PMID- 11472184 TI - The first transition metal complex of tetrafluorobenzyne: Ir(eta(5) C(5)Me(5))(PMe(3))(eta(2)-C(6)F(4)). PMID- 11472185 TI - Evidence for large structural fluctuations of the photobleached intermediate of photoactive yellow protein in solution. PMID- 11472186 TI - Reversible [7]-thiahelicene formation using a 1,2-dithienylcyclopentene photochrome. PMID- 11472187 TI - Fluorous Swern reaction. PMID- 11472188 TI - Remarkable electronic effect on rhodium-catalyzed carbonyl additions and conjugated additions with arylmetallic reagents. PMID- 11472189 TI - Direct detection of heat and cold denaturation for partial unfolding of a protein. PMID- 11472191 TI - Chemoselectivity in metal cation mediated hydrolysis of a phosphonoformate diester. PMID- 11472190 TI - RNA-selective coordination complexes identified via dynamic combinatorial chemistry. PMID- 11472192 TI - [4 + 1] Cycloadditions of cyclohexadienes with osmium nitrides. PMID- 11472193 TI - Catalytic asymmetric synthesis of highly functionalized cyclopentenes by a [3 + 2] cycloaddition. PMID- 11472194 TI - A conformationally programmable ligand. PMID- 11472195 TI - Interpenetrated 3D polymeric metal-radical networks built from a tetranitroxide radical and bis(hexafluoroacethylacetonato) manganese(II). PMID- 11472196 TI - An experimental strategy to dramatically reduce the RF power used in cross polarization solid-state NMR spectroscopy. PMID- 11472197 TI - Reactivity of [MoX(eta(3)-allyl)(CO)(2)(N-N)] complexes with simple, nonstabilized carbanions. PMID- 11472198 TI - Highly regio- and enantioselective Pd-catalyzed allylic alkylation and amination of monosubstituted allylic acetates with novel ferrocene P,N-ligands. PMID- 11472199 TI - Catalytic cycle for the asymmetric hydrogenation of prochiral ketones to chiral alcohols: direct hydride and proton transfer from chiral catalysts trans Ru(H)(2)(diphosphine)(diamine) to ketones and direct addition of dihydrogen to the resulting hydridoamido complexes. PMID- 11472200 TI - Palladium-catalyzed enantioselective oxidations of alcohols using molecular oxygen. PMID- 11472202 TI - Potent and selective carboxylic acid-based inhibitors of matrix metalloproteinases. PMID- 11472203 TI - Experimental precedent for the need to involve the primary hydration layer of DNA in lead drug design. PMID- 11472204 TI - A rational approach to the design of selective substrates and potent nontransportable inhibitors of the excitatory amino acid transporter EAAC1 (EAAT3). new glutamate and aspartate analogues as potential neuroprotective agents. PMID- 11472205 TI - Novel selective PDE4 inhibitors. 1. Synthesis, structure-activity relationships, and molecular modeling of 4-(3,4-dimethoxyphenyl)-2H-phthalazin-1-ones and analogues. AB - A number of 6-(3,4-dimethoxyphenyl)-4,5-dihydro-2H-pyridazin-3-ones and a novel series of 4-(3,4-dimethoxyphenyl)-2H-phthalazin-1-ones were prepared and tested on the cGMP-inhibited phosphodiesterase (PDE3) and cAMP-specific phosphodiesterase (PDE4) enzymes. All tested compounds were found to specifically inhibit PDE4 except for pyridazinone 3b, which showed moderate PDE4 (pIC(50) = 6.5) as well as PDE3 (pIC(50) = 6.6) inhibitory activity. In both the pyridazinone and phthlazinone series it was found that N-substitution is beneficial for PDE4 inhibition, whereas in the pyridazinone series it also accounts for PDE4 selectivity. In the phthalazinone series, the cis-4a,5,6,7,8,8a hexahydrophthalazinones and their corresponding 4a,5,8,8a-tetrahydro analogues showed potent PDE4 inhibitory potency (10/11c,d: pIC(50) = 7.6-8.4). A molecular modeling study revealed that the cis-fused cyclohexa(e)ne rings occupy a region in space different from that occupied by the other fused (un)saturated hydrocarbon rings applied; we therefore assume that the steric interactions of these rings with the binding site play an important role in enzyme inhibition. PMID- 11472206 TI - Novel selective PDE4 inhibitors. 2. Synthesis and structure-activity relationships of 4-aryl-substituted cis-tetra- and cis-hexahydrophthalazinones. AB - A series of 4-aryl-substituted cis-4a,5,8,8a-tetra- and cis-4a,5,6,7,8,8a hexahydro-2H-phthalazin-1-ones with high inhibitory activity toward cAMP-specific phosphodiesterase (PDE4) was synthesized. To study structure-activity relationships various substituents were introduced to the 2-, 3-, and 4-positions of the 4-phenyl ring. Substitution at the 4-position of the phenyl ring was restricted to a methoxy group, probably due to unfavorable steric interactions of larger groups with the binding site. The introduction of many alkoxy substituents including distinct ring systems and functional groups was allowed to the 3 position. It was found that in general the cis-4a,5,8,8a-tetrahydro-2H-phthalazin 1-ones are more potent than their hexahydrophthalic counterparts, the best activity residing in (4-imidazol-1-yl-phenoxy)butoxy analogue 16o (pIC(50) = 9.7). PMID- 11472207 TI - Design, synthesis, DNA binding, and biological evaluation of water-soluble hybrid molecules containing two pyrazole analogues of the alkylating cyclopropylpyrroloindole (CPI) subunit of the antitumor agent CC-1065 and polypyrrole minor groove binders. AB - We have synthesized and evaluated a series of hybrids, denoted 22--27, for in vitro cytotoxic activity against a variety of cancer cell lines. These hybrids represent a molecular combination of polypyrrole minor groove binders structurally related to the natural antitumor agent distamycin A and two pyrazole analogues of the left-hand segment called cyclopropylpyrroloindole (CPI) of the potent antitumor antibiotic (+)-CC-1065. These novel water-soluble hybrids have been designed to enhance the minor groove binding ability of alkylating units 20 and 21, which should increase their clinical appeal by overcoming the administration problems of (+)-CC-1065 derivatives. The DNA alkylating and cytotoxic activities against several tumor cell lines are reported and discussed in terms of their structural differences in relation to both the number of N methyl pyrrole rings and the type of the alkylating unit tethered to the oligopeptidic frame. It may be noted that, in general, and especially for 22--24, the cytotoxicity of the hybrids was much greater than that of the alkylating units alone. In only one case, compound 27, did the hybrid have cytotoxic activity comparable to that of the alkylating unit alone against FM3A/0 cells. The broadest spectrum of activity and greatest potency was shown by the hybrid 24, in which the alkylating unit 20 and the deformyl distamycin A are tethered by 1-methyl 2,5-dicarbonyl pyrazole, with IC(50) values for the different tumor cell lines ranging from 7 to 71 nM. For compounds 22--24, the increase of the length of the pseudopeptidic moiety from one to three N-methylpyrrole residues led to an increased cytotoxicity. Among the hybrids tested for their inhibitory effects on the proliferation of murine L1210 leukemia cell line, compound 24 proved to be the most active (IC(50) = 7.4 nM), and in the sequencing gel experiments, it showed the strongest and most highly sequence-specific DNA alkylation activity. For compounds 22-24, the sequence specificity of DNA alkylation appears to be affected by the modification of the number of pyrrole rings, and the correlation between cytotoxicity and alkylation pattern suggests that 24 exerts its cytotoxicity through DNA sequence-specific alkylation of the third adenine located in the sequence 5'-ACAAAAATCG-3'. The two other hybrids 22 and 23 were slightly less active for tumor cell proliferation, with IC(50) values of 58 and 19 nM, respectively. With only one exception, none of the compounds was endowed with antiviral activity at subtoxic concentrations. Compound 24 inhibited the effect of vaccinia virus at a concentration that was significantly lower than its minimum cytotoxic concentration for the E(6)SM host cells. These compounds gave distinct patterns of alkylation in AT-rich sequences, indicating that minor structural changes produced marked alterations in sequence selectivity. PMID- 11472208 TI - Structure-based design, synthesis, and biological evaluation of conformationally restricted novel 2-alkylthio-6-[1-(2,6-difluorophenyl)alkyl]-3,4-dihydro-5 alkylpyrimidin-4(3H)-ones as non-nucleoside inhibitors of HIV-1 reverse transcriptase. AB - 5-Alkyl-2-(alkylthio)-6-(2,6-difluorobenzyl)-3,4-dihydropyrimidin-4(3H)-ones (S DABOs, 2) have been recently described as a new class of human immunodeficiency virus type 1 (HIV-1) non-nucleoside reverse transcriptase (RT) inhibitors (NNRTIs) active at nanomolar concentrations (Mai, A. et al. J. Med. Chem. 1999, 42, 619-627). In pursuing our lead optimization efforts, we designed novel conformationally restricted S-DABOs, 3, featuring a methyl at the benzylic carbon (Y = Me) and at the pyrimidine 5-position (R = Me). Conformational analyses and docking simulations suggested that the presence of both methyls would significantly reduce conformational flexibility without compromising, in the R enantiomers, the capability of fitting into the RT non-nucleoside binding pocket. To develop structure-activity relationships, we prepared several congeners of type 3 belonging to the thymine (R = Me) and uracil (R = H) series, featuring various 2-alkylthio side chains (X = Me, i-Pr, n-Bu, i-Bu, s-Bu, c-pentyl, and c hexyl) and aryl moieties different from the 2,6-difluorophenyl (Ar = phenyl, 2,6 dichlorophenyl, 1-naphthyl). Moreover, alpha-ethyl derivatives (Y = Et) were included in the synthetic project in addition to alpha-methyl derivatives (Y = Me). All of the new compounds were evaluated for their cytotoxicity and anti-HIV 1 activity in MT-4 cells, and some of them were assayed against highly purified recombinant wild-type HIV-1 RT using homopolymeric template primers. The results were expressed as CC(50) (cytotoxicity), EC(50) (anti-HIV-1 activity), SI (selectivity, given by the CC(50)/EC(50) ratio), and IC(50) (RT inhibitory activity) values. In the 2,6-difluorobenzylthymine (R = Me) series, methylation of the benzylic carbon improved anti-HIV-1 and RT inhibitory activities together with selectivity. Compound 3w (Ar = 2,6-F(2)-Ph, R = Y = Me, X = c-pentyl) turned out the most potent and selective among the S-DABOs reported to date (CC(50) > 200 microM, EC(50) = 6 nM, IC(50) = 5 nM, and SI > 33 333). Assays performed on the pure enantiomer (+)-3w, much more active than (-)-3w, yielded the following results: CC(50) > 200 microM, EC(50) = 2 nM, IC(50) = 8 nM, and SI > 100 000, under conditions wherein MKC-442 was less active and selective (CC(50) > 200 microM, EC(50) = 30 nM, IC(50) = 40 nM, SI > 6666). The 2,6 difluorophenylethylthymines (R = Me) were generally endowed with higher potency compared with the uracil counterparts (R = H). In the 2,6-difluorophenyl series the best and the least performant 2-alkylthio side chains were the 2-c-pentylthio and the 2-methylthio, respectively. When the methyl at the benzylic carbon was replaced by an ethyl, activity was retained or decreased slightly, thus suggesting that the dimensions of the cavity within the RT hosting this substituent would not be compatible with groups larger than ethyl. Aryl moieties different from the 2,6-difluorophenyl (phenyl, 1-naphthyl, 2,6-dichlorophenyl) were generally detrimental to activity, consistent with a favorable electronic effect exerted by the 2,6-fluorines on a putative charge-transfer interaction between the aromatic moieties of the inhibitor and Tyr188. PMID- 11472209 TI - Structural studies on bioactive compounds. 34. Design, synthesis, and biological evaluation of triazenyl-substituted pyrimethamine inhibitors of Pneumocystis carinii dihydrofolate reductase. AB - The triazenyl-pyrimethamine derivative 3a (TAB), a potent and selective inhibitor of Pneumocystis carinii DHFR, was selected as the starting point for a lead optimization study. Molecular modeling studies, corroborated by a recent crystal structure determination of the ternary complex of P. carinii DHFR--NADPH bound to TAB, predicted that modifications to the acetoxy residue of the lead inhibitor could exploit binding opportunities in the vicinity of an active site pocket bounded by residues Ile33, Lys37, and Leu72. Substitutions in the benzyl moiety with electron-donating and electron-withdrawing groups were predicted to probe face-edge interactions with amino acid Phe69 unique to the P. carinii enzyme. New triazenes 10a--v and 12a--f were prepared by coupling the diazonium tetrafluoroborate salt 6b of aminopyrimethamine with substituted benzylamines or phenethylamines. The most potent of the new inhibitors against P. carinii DHFR was the naphthylmethyl-substituted triazene 10t (IC(50): 0.053 microM), but a more substantial increase in potency against the rat liver DHFR led to a reduction in selectivity (ratio rat liver DHFR IC(50)/P. carinii DHFR IC(50): 5.36) compared to the original lead structure 3a (ratio rat liver DHFR IC(50)/P. carinii DHFR IC(50): 114). PMID- 11472210 TI - Disabling erbB receptors with rationally designed exocyclic mimetics of antibodies: structure-function analysis. AB - Overexpression of the HER2 receptor is observed in about 30% of breast and ovarian cancers and is often associated with an unfavorable prognosis. We have recently designed an anti-HER2 peptide (AHNP) based on the structure of the CDR H3 loop of the anti-HER2 rhumAb 4D5 and showed that this peptide can mimic some functions of rhumAb 4D5. The peptide disabled HER2 tyrosine kinases in vitro and in vivo similar to the monoclonal antibody (Park, B.-W. et al. Nat. Biotechnol. 2000, 18, 194--198). AHNP has been shown to selectively bind to the extracellular domain of the HER2 receptor with a submicromolar affinity in Biacore assays. In the present paper, we demonstrate that in addition to being a structural and functional mimic of rhumAb 4D5, AHNP can also effectively compete with the antibody for binding to the HER2 receptor indicating a similar binding site for the peptide and the parental antibody. To further develop AHNP as an antitumor agent useful for preclinical trials and as a radiopharmaceutical to be used for tumor imaging, a number of derivatives of AHNP have been designed. Structure- function relationships have been studied using surface plasmon resonance technology. Some of the AHNP analogues have improved binding properties, solubility, and cytotoxic activity relative to AHNP. Residues in the exocyclic region of AHNP appear to be essential for high-affinity binding. Kinetic and equilibrium analysis of peptide-receptor binding for various AHNP analogues revealed a strong correlation between peptide binding characteristics and their biological activity. For AHNP analogues, dissociation rate constants have been shown to be better indicators of peptide biological activity than receptor binding affinities. This study demonstrates a possibility of mimicking the well documented antibody effects and its applications in tumor therapy by much smaller antibody-based cyclic peptides with potentially significant therapeutic advantages. Strategies used to improve binding properties of rationally designed AHNP analogues are discussed. PMID- 11472211 TI - Original 2-alkylamino-6-halogenoquinazolin-4(3H)-ones and K(ATP) channel activity. AB - A series of 6-substituted 2-alkylaminoquinazolin-4(3H)-ones structurally related to 3-alkylamino-4H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-dioxides were synthesized and tested as putative K(ATP) channel openers on isolated pancreatic endocrine tissue as well as on isolated vascular, intestinal, and uterine smooth muscle. Most of the 6-halogeno-2-alkylaminoquinazolin-4(3H)-ones were found to inhibit insulin release from pancreatic B-cells and to exhibit vasorelaxant properties. In contrast to their pyridothiadiazine dioxide isosteres previously described as more active on the endocrine than on the smooth muscle tissue, quinazolinones cannot be considered as tissue selective compounds. Biological investigations, including measurements of (86)Rb, (45)Ca efflux from pancreatic islet cells and measurements of vasodilator potency in rat aortic rings exposed to 30 or 80 mM KCl in the presence or the absence of glibenclamide, were carried out with 6 chloro- and 6-iodo-3-isopropylaminoquinazolin-4(3H)-ones. Such experiments showed that, depending on the tissue, these new compounds did not always express the pharmacological profile of pure K(ATP) channel openers. Analyzed by X-ray crystallography, one example of quinazolinones appeared to adopt a double conformation. This only suggests a partial analogy between the 2 alkylaminoquinazolin-4(3H)-ones and the 3-alkylamino-4H-pyrido[4,3-e]-1,2,4 thiadiazine 1,1-dioxides. In conclusion, the newly synthesized quinazolinones interfere with insulin secretion and smooth muscle contractile activity. Most of the compounds lack tissue selectivity, and further investigations are required to fully elucidate their mechanism(s) of action. PMID- 11472212 TI - Design and synthesis of potent and selective alpha(4)beta(7) integrin antagonists. AB - Interactions of the integrins alpha(4)beta(7) with its cognate ligand mucosal addressin cell adhesion molecule-1 (MAdCAM-1) play a crucial role in the development of mucosa-associated lymphoid organs, in the generation of mucosal immune responses, and in diverse pathological processes such as chronic inflammatory bowel disease and type I diabetes. Using a previously developed spatial screening technique we describe the development of potent and selective alpha(4)beta(7) integrin antagonists based on the domain 1 Leu-Asp-Thr (LDT) sequence of MAdCAM-1 that is essential for alpha(4)beta(7) integrin binding. A library of homodetic cyclic penta- and hexapeptides was synthesized presenting the pharmacophoric LDT-sequence in different conformations. The cyclic hexapeptide P10 cyclo(Leu-Asp-Thr-Ala-D-Pro-Ala) inhibits alpha(4)beta(7) integrin mediated cell adhesion to MAdCAM-1 effectively. Further optimization of the lead structure P10 resulted in cyclic hexapeptides with enhanced activity. The compounds P25 cyclo(Leu-Asp-Thr-Ala-D-Pro-Phe), P28 cyclo(Leu-Asp-Thr-Asp-D Pro-Phe), P29 cyclo(Leu-Asp-Thr-Asp-D-Pro-His), and P30 cyclo(Leu-Asp-Thr-Asp-D Pro-Tyr) strongly inhibited alpha(4)beta(7) integrin mediated cell adhesion to MAdCAM-1, but they did not affect binding of the closely related alpha(4)beta(1) integrin to VCAM-1. PMID- 11472213 TI - Structure-activity studies of ground- and transition-state analogue inhibitors of cyclophilin. AB - Peptidyl-prolyl isomerases (PPIases) are ubiquitous cellular enzymes that play roles in cellular signaling and protein folding. In addition, these proteins are the receptors for the widely used immunosuppressants cyclosporin A and FK506. We report the first structure-activity studies of de novo designed inhibitors of cyclophilin, the cellular target of cyclosporin A. Our mechanism-based inhibitors were modeled on the ground- and transition-state structures of proline-containing peptides, the natural substrates of the enzyme. Both ground-state analogues 1 and transition-state analogues 2 were prepared as single enantiomers from L-proline following a "self-reproduction of chirality" procedure. The binding affinities of the analogues for the active site of cyclophilin were measured by a fluorescence perturbation assay. While the transition-state analogues 2 did not display significant avidity for the active site (K(d) = 77 microM for 2b), several ground state analogues bound to the enzyme with low micromolar affinity (K(d) = 1.5 microM for 1e). These results proclaim that properly designed small molecular weight molecules can form strong complexes with cyclophilin and may find use as probes in cell biology and as therapeutic agents. PMID- 11472214 TI - The effect of 1,3-diaryl-[1H]-pyrazole-4-acetamides on glucose utilization in ob/ob mice. AB - This article provides evidence of a new class of compounds, 1,3-diaryl-[1H] pyrazole-4-acetamides, initially identified from their ability to increase glucose transport in an adipocyte and muscle cell line and ultimately demonstrating dramatic glucose lowering in ob/ob mice, a diabetic animal model. The lead compound, 1, possessed some behavioral-like effects which were removed by structural variation during the course of this investigation. Specifically, 11g (R1 = meta-CF(3), Ar2 = 4'biphenyl, R3 = diethylamide) illustrated the potency of this series with ED(50) values for glucose lowering in ob/ob mice of 3.0 mg/kg/day. Concomitant with its effect on glucose lowering, 11g also caused a 50% reduction in insulin levels consistent with an agent that increases whole body insulin sensitivity. 11g showed favorable pharmacokinetic data with acceptable absorption, negligible metabolism, and good duration of action. 11g demonstrated no appreciable adipogenic effect through PPAR gamma agonism, a characteristic of the thiazolidinediones (TZD), and so represents a potentially new class of agents for the treatment of diabetes. PMID- 11472215 TI - Synthesis of cytotoxic 6E-hydroximino-4-ene steroids: structure/activity studies. AB - In an effort to determine the pharmaceutical utility and the structural requirements for activity against various tumor cell lines, several 6E hydroximino-4-ene steroids with different side chains and degrees of unsaturation on ring A were synthesized in our laboratory. Evaluation of the synthesized compounds for cytotoxicity against P-388, A-549, HT-29, and MEL-28 tumor cells revealed that some important structural features are required for activity. The presence of a cholesterol-type side chain, which appears to play a major role in determining the biological activity, the existence of a ketone functionality at C 3, and an elevated degree of oxidation on ring A all result in higher bioactivity than other structural motifs. PMID- 11472216 TI - Synthesis of 6- and 7- hydroxy-8-azabicyclo[3.2.1]octanes and their binding affinity for the dopamine and serotonin transporters. AB - Cocaine is a potent stimulant of the central nervous system. Its reinforcing and stimulant effects are related to its ability to inhibit the membrane bound dopamine transporter (DAT). Inhibition of the DAT causes an increase of dopamine in the synapse with a resultant activation of postsynaptic receptors. The rapid onset and short duration of action of cocaine contribute to its high addictive potential. Consequently, the design of tropane analogues of cocaine that display longer onset times on the DAT and extended duration of action is driven by the need to develop cocaine medication. This study extends the exploration of bridge hydroxylated azabicyclo[3.2.1]octanes (tropanes). A series of 6- and 7 hydroxylated tropanes was prepared and evaluated biologically. Structure activity relationships lead to the following conclusions. Bridge hydroxylated tropanes retain biological enantioselectivity but display higher DAT versus SERT selectivity, particularly for the 3 alpha-aryl compounds as compared with the 3beta-aryl compounds, than the bridge unsubstituted analogues. The 7-hydroxyl compounds are more potent at the DAT than their 6-hydroxyl counterparts. The general SAR of the tropanes is maintained and the rank order of potencies based on substitution at the C3 position remains 3,4-dichloro > 2-naphthyl > 4-fluoro > phenyl. PMID- 11472217 TI - Design, synthesis, and structure-activity relationships of macrocyclic hydroxamic acids that inhibit tumor necrosis factor alpha release in vitro and in vivo. AB - To search for TNF-alpha (tumor necrosis factor alpha) converting enzyme (TACE) inhibitors, we designed a new class of macrocyclic hydroxamic acids by linking the P1 and P2' residues of acyclic anti-succinate-based hydroxamic acids. A variety of residues including amide, carbamate, alkyl, sulfonamido, Boc-amino, and amino were found to be suitable P1-P2' linkers. With an N-methylamide at P3', the 13-16-membered macrocycles prepared exhibited low micromolar activities in the inhibition of TNF-alpha release from LPS-stimulated human whole blood. Further elaboration in the P3'-P4' area using the cyclophane and cyclic carbamate templates led to the identification of a number of potent analogues with IC(50) values of 0.05 (n=6). No significant change was observed when thiopental was administered to the cranial circulation: low dose, 1061+/-1167 to 965+/-874 impulses/min at 1 min post-injection, P>0.05 (n=10); high dose, 864+/-331 to 917+/-525 impulses/min at 1 min post-injection, P>0.05 (n=8). CONCLUSION: Thiopental has a direct (spinal) depressant effect on dorsal neuronal responses to noxious stimulus, but no significant supraspinal effect. PMID- 11472282 TI - Effects of combined methohexitone-remifentanil anaesthesia in electroconvulsive therapy. AB - BACKGROUND: Methohexitone is widely used to provide anaesthesia for patients undergoing electroconvulstive therapy (ECT). Short seizure duration, high blood pressures (BP) and heart rates (HR) are usual in elderly patients. In this study, elderly patients undergoing ECT received low dose methohexitone with remifentanil or methohexitone alone and motor seizure duration, haemodynamic response and recovery time were compared. METHODS: Ten patients, of mean age 74.3 years, were enrolled in this double-blind, randomised crossover trial, receiving a total of 38 ECTs. Each patient was given the following two i.v. regimens in random order: A) methohexitone 0.5 mg kg(-1) combined with remifentanil 1.0 microg kg(-1) and B) methohexitone 0.75 mg kg(-1). Additional methohexitone was given, if needed, until loss of consciousness, and then suxamethonium 1.0 mg kg(-1) for muscular paralysis. RESULTS: Mean motor seizure duration was significantly longer with methohexitone-remifentanil (37.6 s (SD 12.0)) than with methohexitone alone (27.1 (SD11.5)) (P=0.0009). Recovery time, time to spontaneous breathing, peak postictal changes in BP and HR were similar with both regimens. CONCLUSION: A reduced dose of methohexitone combined with remifentanil allows prolonged duration of motor seizures in ECT. We conclude that low dose methohexitone combined with a short-acting opioid is a reasonable alternative for elderly patients undergoing ECT, and for other patients with short seizure duration. PMID- 11472283 TI - Being awake intermittently during propofol-induced hypnosis: a study of BIS, explicit and implicit memory. AB - BACKGROUND: Being awake during anaesthesia is a serious complication. An anaesthetic depth monitor must discriminate in real time between wakefulness and unconsciousness. The present study created a period of wakefulness during propofol-induced hypnosis. Bispectral index (BIS), explicit and implicit memories of the awake period were investigated. METHODS: Ten volunteers were studied. The calculated brain concentration of a target controlled infusion of propofol was increased until loss of response (LOR) to verbal command and then propofol was stopped. When fully awake, volunteers were presented with a picture, sound and smell. Propofol infusion was restarted until LOR and then ceased. BIS and the calculated brain concentration of propofol were recorded every minute. A structured interview was conducted for explicit memories after awakening and for explicit as well as implicit memories the day after. RESULTS: Median BIS-index for the transition between awake and asleep and vice versa differed significantly. It was not possible, however, to establish any threshold value or zone for discriminating between wakefulness and LOR due to the large inter individual variations in BIS-index. No volunteer could explicitly recall any of the stimuli presented during the period of wakefulness. CONCLUSION: The BIS-index decreases with increasing sedation but because of the large individual variations, the real-time BIS-index for the individual subject cannot reliably discriminate wakefulness from unconsciousness during propofol infusion. Propofol causes such profound amnesia that lack of postoperative recall does not assure that episodes of awareness have not occurred during propofol-induced hypnosis. PMID- 11472284 TI - Pain on injection of propofol with or without infusion of carrier fluid. AB - BACKGROUND: Propofol, a popular intravenous (iv) anaesthetic induction agent for brief cases or day surgery, is associated with smooth induction, pleasant sleep, rapid recovery and little postoperative nausea. A major disadvantage is pain at the site of injection. The aim of the present study was to examine the influence of simultaneous iv infusion of carrier fluid on propofol-induced local pain. METHODS: Thirty patients, scheduled for ear-nose-throat or plastic surgery under general anaesthesia, were randomly allocated into two groups. Each patient had two 2 ml iv bolus injections of propofol given at two minutes' interval. In group I (n=15) the first bolus injection was given with no iv carrier fluid and the second one given with a 10 ml iv carrier fluid infused over 10 s. Correspondingly, the patients in group II (n=15) had their first injection with and their second one without the iv carrier fluid. Following each injection of propofol the patients were asked by a blinded investigator to score their pain on a 10-point visual analogue scale, and to report the appearance, maximum and disappearance of pain. After the second assessment of pain, general anaesthesia was induced with more propofol. RESULTS: Pain intensity at the site of propofol injection was found not to be influenced by simultaneous iv infusion of carrier fluid. CONCLUSION: It seems, from the results obtained here, that simultaneous iv infusion of carrier fluid has no particular effect on local pain following iv administration of propofol. PMID- 11472285 TI - Controlled hypercapnia during one-lung ventilation in patients undergoing pulmonary resection. AB - BACKGROUND: In a pilot study, the haemodynamic and gas exchange effects of acute hypercapoia during one-lung ventilation in thoracotomy patients were investigated. The effects of normocapnic one-lung ventilation (OLV-N) on haemodynamics and pulmonary gas exchange were compared with those of hypercapnic one-lung ventilation (OLV-H) in 14 patients undergoing pulmonary lobectomy. METHODS: Hypercapnia was induced by decreasing tidal volume until PaCO2 increased to 8-9 kPa. During OLV, minute ventilation was reduced from 8.8+/-1.7 to 4.2+/ 0.7 l min(-1). RESULTS: Cardiac index (from 3.3+/-0.6 to 3.9+/-0.6 l min(-1), P<0.01) and pulmonary vascular resistance index (from 245+/-96 to 347+/-125 dyn s cm(-5) m(-2), P<0.05) increased during OLV-H, whereas systemic vascular resistance index decreased from 1952+/-403 to 1636+/-361 dyn s cm(-5) m(-2) (P<0.01). Pulmonary oxygenation remained unchanged. CONCLUSIONS: All patients had an uneventful course during OLV-H. The determinants of pulmonary oxygenation during hypercapnic one-lung ventilation remain to be further elucidated. PMID- 11472286 TI - High-dose flumazenil potentiates the hypnotic activity of propofol, but not that of thiopental, in ddY mice. AB - BACKGROUND: Flumazenil is a specific benzodiazepine agonist, which is reported to have a partial benzodiazepine agonist-like effect at a high dose. This study investigated the effects of flumazenil and midazolam on the hypnotic dose of propofol and thiopental in ddY mice, using a behavioral model. METHODS: Mice were given either propofol or thiopental intravenously to induce hypnosis, which was defined as a loss of the righting reflex. The mice were pre-treated with flumazenil (0.05, 0.1, or 0.2 mg kg(-1)) or midazolam (0.1 or 0.2 mg kg(-1)), and given propofol or thiopental after a 30-s delay. RESULTS: Pre-treatment with flumazenil (0.1 or 0.2 mg kg(-1)) significantly decreased the hypnotic dose of propofol compared to the control group (9.3+/-0.39 [8.5-10.0] or 9.0+/-0.28 [8.5 9.6] vs. 10.8+/-0.42 [9.9-11.6] mg kg(-1) (ED50+/-SEM and [95% confidence interval]) P<0.05), but not that of thiopental (9.1+/-0.30 [8.5-9.7] with 0.2 mg kg(-1) flumazenil vs. 9.3+/-0.42 [8.4-10.1] mg kg(-1) with saline). Midazolam reduced the hypnotic dose of both propofol and thiopental. Flumazenil antagonized the potentiating effect of midazolam (0.2 mg kg(-1)) on the hypnotic activity of propofol. CONCLUSIONS: These results suggest that the hypnotic activity of propofol is potentiated by the partial agonist activity of flumazenil in ddY mice. PMID- 11472287 TI - Determinants of ICU mortality in necrotizing pancreatitis: the influence of Staphylococcus epidermidis. AB - BACKGROUND: The severity of acute necrotizing pancreatitis ranges from self limited to rapidly progressive illness leading to multiple organ failure. Several scoring systems and clinical parameters have been used to predict the course of the disease. The aim of this study was to evaluate the clinical and microbiological determinants of poor outcome in necrotizing acute pancreatitis. METHODS: Medical records of 67 consecutive patients admitted to the intensive care unit (ICU) of Oulu University Hospital due to acute necrotizing pancreatitis were retrospectively analyzed. All patients received standard surgical intensive care. RESULTS: Patients who died (n=14) had significantly higher APACHE II, SAPS II and Ranson scores at admission to the ICU and maximum SOFA score achieved during ICU stay than did the survivors. The non-survivors were hospitalized later from the time the symptoms were first manifest (5.3 vs. 2.4 days, P=0.051). Mechanical ventilation (P=0.002), surgical management (P=0.028), open packing surgical management (P=0.03), renal replacement therapy (P<0.001), use of inotropic drugs (P=0.012) and Staphylococcus epidermidis growth (P=0.029) in infected pancreatic tissue were all associated with mortality. CONCLUSIONS: In this study the time to hospitalization, severity of illness, intensity of care, and surgical management were associated with poor outcome. In addition, Staphylococcus epidermidis in pancreatic necrosis was associated with increased mortality. PMID- 11472288 TI - Time course of changes in jugular venous oxygen saturation during hypothermic or normothermic cardiopulmonary bypass in patients with diabetes mellitus. AB - BACKGROUND: Preexisting diabetic mellitus is a risk factor determining postoperative neurological disorders. The present study assesses the effects of normothermic and hypothermic cardiopulmonary bypass (CPB) on jugular venous oxygen saturation (SjvO2)in patients with preexisting diabetic mellitus. METHODS: Sixteen diabetic patients who underwent elective coronary artery bypass grafting surgery were randomly divided into two groups: Group DN (n=8, diabetic patients) underwent normothermic CPB (>35 degrees C), and group DH (n=8, diabetic patients) underwent hypothermic CPB (32 degrees C). Controls were 16 age-matched non diabetic patients (normothemic group, CN: n=8; hypothemic group, CH: n=8). A 4.0 F fiberoptic oximetry oxygen saturation catheter was inserted into the right jugular bulb to continuously monitor SjvO2 values. Hemodynamic parameters and arterial and jugular venous blood gases were measured seven times. RESULTS: Cerebral desaturation, which was defined as SjvO2 values below 50%, was observed during normothermic CPB in diabetic patients (at the onset of CPB: 46+/-3%, at 20 min after onset of CPB: 49+/-3%, means+/-SD, respectively). No cerebral desaturation occurred in diabetic and control patients during hypothermic CPB. CONCLUSIONS: Patients with preexisting diabetes mellitus experienced cerebral desaturation during normothermic CPB. PMID- 11472289 TI - Changes in cardiac output and intrathoracic blood volume: a mathematical coupling of data? AB - BACKGROUND: Measurements of intrathoracic blood volume (ITBV) provide volumetric information about cardiac preload and are used to investigate the cause of alterations in cardiac output (CO). On the other hand, CO is required to calculate ITBV. Thus, concerns have been raised with respect to a mathematical coupling of data. The aim of this prospective, clinical study was to investigate whether a variation in CO induced by high-dose beta-blockade influences thermodilution measurements of ITBV in the absence of changes in intravascular volume in patients undergoing minimally invasive coronary artery bypass grafting. METHODS: Sixteen patients undergoing elective minimally invasive direct coronay artery bypass (MIDCAB) surgery were studied. Transpulmonary thermodilution measurements of ITBV and CO were simultaneously performed before bypass grafting, during beta-blockade induced by high-dose esmolol and at the end of surgery. RESULTS: During esmolol administration, CO significantly decreased by 33%, whereas ITBV remained unchanged compared to control values (876+/-46 ml m-2 during control versus 860+/-61 ml m-2 during esmolol administration). After the end of esmolol administration, CO significantly increased by 79%. Again, ITBV remained virtually unchanged (860+/-61 ml m-2 during esmolol administration versus 911+/-38 ml m-2 after esmolol administration). CONCLUSIONS: The results of the present study demonstrate that substantial alterations in CO as a consequence of high-dose esmolol infusion are not associated with changes in ITBV. Because haemodynamic changes were induced by factors other than variation of preload, these findings suggest that changes in cardiac output do not influence thermodilution measurements of ITBV in this setting. PMID- 11472290 TI - Analysis of left ventricular systolic function during elevated external cardiac pressures: an examination of measured transmural left ventricular pressure during pressure-volume analysis. AB - BACKGROUND: Variations or disturbances in intrathoracic and extracardiac pressures (ECP) occur in critically ill and anaesthetised patients. There are uncertainties concerning the analysis of left ventricular pressure-volume relationship (LVPVR) and the calculation of systolic function parameters when conducted without reference to transmural left ventricular pressure (LVPtm) in the setting of elevated ECP. METHODS: In 7 anaesthetised adult pigs, we measured LVPVR using conductance volumetry and tip manometry along with measurement of pericardial and other intrathoracic pressures. Experimental pericardial infusion and pleural insufflation were performed. Transient controlled preload reductions were accomplished using balloon occlusion of the inferior vena cava. Preload recruitable stroke work (PRSW) was calculated using both intracavitary left ventricular pressure (LVPic) and LVPtm, and differences were tested for using a paired t-test. RESULTS: The pericardial and pleural interventions produced significant elevations in ECP. No difference in PRSW calculated using LVPic and LVPtm was detected. CONCLUSION: These results suggest that LVPtm need not be measured and included in LVPVR analysis of systolic function when there is significant external cardiac pressure. To be able to employ LVPVR analysis of systolic function without reference to LVPtm is important for simplified application in the clinical setting, particularly when elevated extracardiac pressures are suspected, or have been therapeutically induced, as with continuous positive pressure ventilation. PMID- 11472291 TI - Effects of positive end-expiratory pressure on intestinal circulation during graded mesenteric artery occlusion. AB - BACKGROUND: Reduced gut perfusion is associated with multiple organ failure. Positive end-expiratory pressure (PEEP) reduces cardiac output (CO) and portal blood flow, and might be detrimental in a situation of already compromised intestinal circulation. The aim of this study was to investigate regional circulatory and metabolic effects of PEEP during graded regional hypoperfusion. METHODS: In 12 barbiturate-anesthetized pigs, we measured systemic and regional blood flows (superior mesenteric arterial, QSMA and portal venous, QPORT), jejunal mucosal perfusion (LDF), tissue oxygenation (PO2TISSUE) and metabolic parameters at PEEP (0, 4, 8 and 12 cm H2O) in a randomized order. Measurements were performed at unrestricted intestinal perfusion pressures (IPP) and at IPP levels of 50 and 30 mmHg. RESULTS: During unrestricted IPP, PEEP decreased MAP, CO, QSMA and QPORT, while systemic, and preportal (RPORT) vascular resistances and jejunal mucosal perfusion were not significantly changed. Preportal tissue oxygen delivery and PO2TISSUE decreased, while preportal tissue oxygen uptake was unaltered. During restricted IPP, PEEP produced the same pattern of hemodynamic alterations as when IPP was not restricted. QPORT and QSMA were lowered by the reductions in IPP, and QPORT was further reduced during PEEP. At an IPP of 30 mmHg, this reduction in QPORT decreased preportal tissue oxygen uptake. Consequently, intestinal ischemia, as indicated by increased net lactate production, occurred. Simultaneously, jejunal mucosal perfusion and PO2TISSUE declined. CONCLUSION: At IPP levels below 50 mmHg, even moderate levels of PEEP impaired local blood flow enough to cause intestinal ischemia. Our data underscore the importance of considering regional circulatory adaptations during PEEP ventilation. PMID- 11472292 TI - Interactions of halothane with isoproterenol and epinephrine on canine epicardial conduction velocity at normal and elevated potassium levels. AB - BACKGROUND: Halothane is known to potentiate catecholamine-induced depression of conduction velocity in Purkinje fibers but not endocardial muscle fibers. The purpose of this study was to examine the interactions of halothane with epinephrine and isoproterenol on canine epicardial conduction velocity at moderately elevated extracellular potassium concentration ([K]0). METHODS: Epicardial muscle strips (10x10x2 mm) were superfused with Tyrode's solution containing 4 or 8 mM [K]0 in the presence of 5 microM epinephrine or 1 microM isoproterenol with or without 0.8 mM halothane. Conduction velocity in the longitudinal and transverse directions relative to epicardial fiber orientation was recorded during alternate stimulation in each direction. RESULTS: In the presence of halothane, a change from 4 to 8 mM [K]0 decreased (P< or =0.05) longitudinal and transverse conduction velocities by 26% and 21%, respectively. Isoproterenol alone at 4 and 8 mM [K]0 depressed (P<0.05) both longitudinal and transverse conduction velocities. However, the depression of longitudinal conduction velocity by isoproterenol at 4 mM [K]0 was attenuated by halothane. Epinephrine with or without halothane depressed (P<0.05) both longitudinal and transverse conduction velocities at 8 but not at 4 mM [K]0. CONCLUSION: The results do not support a synergistic interaction between halothane and epinephrine on myocardial conduction but do demonstrate depression of conduction by epinephrine at 8 mM [K+]0, a potassium ion concentration comparable to those reported following epinephrine infusions. PMID- 11472293 TI - Epidural washout with high volumes of saline to accelerate recovery from epidural anaesthesia. AB - BACKGROUND: Prolonged postoperative blockade can follow neuraxial blocks for short surgical procedures. We investigated whether washout with a high volume of saline through an epidural catheter could provide a faster recovery after epidural anaesthesia. METHODS: Thirty patients were randomly assigned to a control group (no washout), to group 2x (epidural washout with twice the volume of 2% mepivacaine) and group 4x (epidural washout with four times that volume). RESULTS: Recovery times from sensory blockade at L2 were 151+/-24, 122+/-29 and 116+/-24 min for control, 2x and 4x groups respectively. Significant differences were found in both saline groups when compared with control group, but not between group 2x and group 4x. No differences were found concerning motor blockade. One patient in group 4x demonstrated signs of intracranial hypertension. Mepivacaine plasma concentrations were increased by saline washout in group 4x. CONCLUSIONS: Epidural washout with a high volume of saline can not be recommended since no clinically significant reduction in the recovery time can be achieved without risk. PMID- 11472294 TI - High concentrations of adrenergic antagonists prolong sciatic nerve blockade by tetrodotoxin. AB - BACKGROUND: Millimolar-range concentrations of some adrenergic antagonists have been shown to have local anesthetic-like properties, and to stimulate GTPase activity in vitro. In this report, we investigate whether these agents can potentiate the effect of tetrodotoxin (TTX) and bupivacaine, a conventional local anesthetic, and whether GTPase activation plays a role. METHODS: Rats received sciatic nerve blockade with tetrodotoxin or bupivacaine co-injected with adrenergic antagonists and/or agonists, or pertussis toxin. Thermal nociceptive blockade was quantified with modified hotplate testing. RESULTS: Nerve block from TTX alone lasted 153 (99-223) min (median and 25th and 75th percentiles). Co injection with 20 mM phentolamine, propranolol, and yohimbine prolonged TTX block to 856 (765-862), 486 (444-510), and 465 (413-495) min respectively (P<0.005 in all cases, compared to TTX alone). Micromolar concentrations of adrenergic antagonists (which inhibited the prolongation of TTX block by epinephrine) did not prolong TTX block. Injection of adrenergic antagonists alone did not produce specific nerve block. They did not prolong TTX block when injected at a remote subcutaneous site. Prolongation of TTX block by phentolamine was not inhibited by co-injection with pertussis toxin. Adrenergic antagonists did not prolong bupivacaine block. CONCLUSIONS: High concentrations of adrenergic antagonists markedly prolonged TTX block, but not bupivacaine block. This locally mediated action does not appear to be adrenergic-receptor-specific, or mediated by GTPase activation. PMID- 11472295 TI - Perineural antinociceptive effect of opioids in a rat model. AB - BACKGROUND: The research on conductive analgesia induced by perineural opioids generated a large body of conflicting data. In this study we reassessed the antinociceptive response to perineural administration of morphine, fentanyl or meperidine in a rat model. METHODS: Analgesia was assessed using the hind paw withdrawal latency (HPWL) response to radiant heat. The opioid dose producing 20% of maximal possible effect (20%MPE) for systemic analgesia was calculated for each drug. Then sciatic blockade was performed with the dose corresponding to 20%MPE. The injected hind paw was used to measure direct perineural effect and the contralateral hind paw was used as an indicator of systemic effect. RESULTS: The response latency produced by morphine or fentanyl was not significantly different for ipsilateral (perineural effect) or contralateral (systemic effect) paw (27+/-11 vs. 28+/-16 and 3l+/-16 vs. 23+/-16 s, respectively). However, the meperidine group showed significantly higher %MPE for the ipsilateral paw (79+/ 32 s) than for the contralateral paw (27+/-22 s). CONCLUSIONS: The results indicate that perineural fentanyl or morphine do not produce analgesia. Perineural block produced by meperidine was attributed to local anesthetic-like effect, rather than to drug interaction with opioid receptor. PMID- 11472296 TI - Pulmonary hemorrhage in association with negative pressure edema in an intubated patient. AB - Negative pressure pulmonary edema due to upper airway obstruction following extubation is a well-recognized problem. However, frank pulmonary hemorrhage as a manifestation of upper airway obstruction is uncommon. We report a case of significant pulmonary hemorrhage and negative pressure pulmonary edema in an intubated patient. Bronchoscopy showed a collection of blood in the right lower lobe of the lungs, suggesting a localized source of bleeding. There have been two previously reported cases of pulmonary hemorrhage after upper airway obstruction. One suggested that the bleeding was due to damage to the pulmonary capillaries, the other that it was due to disruption of the bronchial vessels. We feel that in our case there was some indication that the pulmonary bleeding was a result of bronchial vessel damage. A number of factors might have been involved in its development, including negative pulmonary pressure, recent respiratory tract infection, and positive airways pressure (due to coughing). PMID- 11472297 TI - Anticoagulation with hirudin for continuous veno-venous hemodialysis in liver transplantation. AB - Postoperative encephalopathy after orthotopic liver transplantation can be difficult to diagnose. We report a case of heparin-induced thrombocytopenia in a liver transplant patient who had seizures. Due to poor liver function the patient developed hepatorenal syndrome requiring continuous veno-venous hemodialysis (CVVHD). Initially we used heparin as the anticoagulant. After the diagnosis of heparin-induced thrombocytopenia (HIT) was made, we switched to r-hirudin. No serious side effects, e.g. bleeding or immune sensitization, were seen. PMID- 11472298 TI - Spinal anesthesia for a patient with type I sialidosis undergoing abdominal surgery. AB - Type I and II sialidosis are autosomal recessively inherited glycoprotein storage disorders. Until now, there has been no published reports of patients with these conditions requiring anesthesia. We present the case of a 31-year-old male afflicted with type I sialidosis who underwent a surgical jejunostomy. Regional (spinal) anesthesia was carried out uneventfully. We discuss the anesthetic challenges posed by patients with type I and II sialidosis. Airway assessment and management is particularly crucial. PMID- 11472299 TI - Is nitrous oxide a real gentleman? PMID- 11472300 TI - Reply Nitrous oxide, a gentleman? Absolutely! PMID- 11472301 TI - Nitrous oxide: an ageing gentleman. PMID- 11472302 TI - Effects of C-peptide on forearm blood flow and brachial artery dilatation in patients with type 1 diabetes mellitus. AB - Recent studies suggest that C-peptide increases blood flow in both exercising and resting forearm in patients with type 1 diabetes. Now we have studied the effect of C-peptide administration on endothelial-mediated and non-endothelial-mediated arterial responses as well as central haemodynamics in 10 patients with type 1 diabetes in a placebo-controlled double-blind study. Euglycaemia was maintained with an i.v. insulin infusion before and during the study. A high-resolution ultrasound technique and Doppler echocardiography were used to assess haemodynamic functions. Brachial artery blood flow and brachial artery diameter were measured in the basal state, 1 and 10 min after reactive hyperaemia and 4 min after sublingual glyceryl trinitrate administration (GTN; endothelial independent vasodilatation), both before and after the end of 60-min C-peptide (6 pmol kg-1 min-1) or saline infusion periods. Echocardiographic measurements were also performed before and at the end of the infusion periods. Seven healthy age matched males served as controls for vascular studies. The patients showed a blunted brachial dilatation after reactive hyperaemia in comparison with the healthy controls (2.1 +/- 0.5% vs. 9.3 +/- 0.3%, P < 0.001), indicating a disturbed endothelial function. C-peptide infusion compared with saline resulted in increased basal blood flow (33 +/- 6%, P < 0.001) and brachial arterial dilatation (4 +/- 1%, P < 0.05). Left ventricular ejection fraction seemed to be improved (5 +/- 2%, P < 0.05) at the end of C-peptide infusion compared with placebo. The vascular response to reactive hyperaemia and GTN was not affected by C-peptide infusion. Our results demonstrate that physiological concentrations of C-peptide increase resting forearm blood flow, brachial artery diameter and left ventricular systolic function in patients with type 1 diabetes. PMID- 11472303 TI - Diffusion-limited tissue equilibration and arteriovenous diffusion shunt describe skeletal muscle nitrous oxide kinetics at high and low blood flows in sheep. AB - This study evaluated the relative importance of perfusion and diffusion mechanisms in compartmental models of blood : tissue inert gas exchange in skeletal muscle. Nitrous oxide kinetics in a hind limb skeletal muscle bed were determined during and after 20 min of nitrous oxide inhalation, at separate low and high steady states of hind limb blood flow in five sheep under halothane anaesthesia. Nitrous oxide concentrations in arterial and femoral vein blood were determined using gas chromatographic analysis and femoral vein blood flow was monitored continuously. Parameters and model selection criteria of various perfusion- or diffusion-limited structural models of skeletal muscle were estimated by simultaneous fitting of the models to the mean observed femoral vein nitrous oxide concentration for both blood flow states. Purely perfusion-limited models fit the data poorly. Models that allowed a diffusion-limited exchange of nitrous oxide between a perfusion-limited tissue compartment and an unperfused deep compartment provided better overall fit of the data and credible parameter estimates. The data was best described by allowing, in addition to diffusion limited tissue equilibration, counter current diffusion of nitrous oxide between arterial and venous blood. The level of tissue blood flow modifies the magnitudes of both these diffusion effects. These results suggest a dual role of diffusion in blood : tissue inert gas equilibration in skeletal muscle. PMID- 11472304 TI - Differential effects of diminished oestrogen and androgen levels on development of skeletal muscle fibres in hypogonadal mice. AB - Androgen and oestrogen hormones influence skeletal muscle size and the characteristics of skeletal muscle fibre types. These effects have typically been assessed by producing acute shortages (castration/ovariectomy) or by hormone supplementation. Little evidence exists, however, on how sex hormone shortages affect muscle development from early stages through to adulthood. Using the hypogonadal mouse model (hpg) we examined the effects of diminished androgen and oestrogen upon muscle size and fibre type composition in murine gastrocnemius and soleus muscles. Hypogonadal male soleus muscle was significantly smaller than normal males, and approximated the normal and hypogonadal females weight and fibre type characteristics. The hypogonadal male gastrocnemius muscle, however, was significantly small in comparison with normal and hypogonadal female gastrocnemius muscles, with the type IIB fibre diameters decreased most markedly. The hypogonadal female soleus muscle approximated the normal female phenotype, but the gastrocnemius muscle was larger than the normal female, approximating the size of the normal male gastrocnemius muscle. Here too, the type IIB fibres showed the most alteration, with greatly increased fibre diameters. Appropriate amounts of androgens were necessary for gender-specific patterns of growth in male muscles, whilst similar amounts of oestrogen were necessary for female gastrocnemius muscle growth, but not for female soleus muscle. Hypogonadism in this murine model generally retards muscle development in males, but has no apparent influence or enhances muscle development in females. Type IIB fibres are most dependent upon sex hormones for appropriate development, but this relationship is muscle-specific. PMID- 11472305 TI - Power and peak blood lactate at 5050 m with 10 and 30 s 'all out' cycling. AB - Anecdotal observations suggest that the reduction in peak lactate accumulation in blood ([La]b peak) after exhausting exercise, in chronic hypoxia vs. normoxia, may be related to the duration of the exercise protocol, being less pronounced after short supramaximal exercise than after incremental exercise (IE) lasting several minutes. To test this hypothesis, six healthy male Caucasians (age 36.8 +/- 7.3, X +/- SD) underwent three exercise protocols on a cycle ergometer, at sea level (SL) and after 21 +/- 10 days at 5050 m altitude (ALT): (1) 10 s, (2) 30 s 'all out' exercise and (3) IE leading to exhaustion in approximately 20-25 min. 'Average' power output (P) was calculated for 10 or 30 s 'all out'; maximal power output (Pmax) was determined for IE. Lactate concentration in arterialized capillary blood ([La]b) was measured at rest and at different times during recovery; the highest [La]b during recovery was taken as [La]b peak. No significant differences in P were observed between SL and ALT, for either 10 or 30 s 'all out' exercise; Pmax during IE was significantly lower at ALT than at SL. [La]b peak after 10 s 'all out' was unaffected by chronic hypoxia (7.0 +/- 0.9 at ALT vs. 6.3 +/- 1.8 mmol x L(-1) at SL). After 30 s 'all out' the [La]b peak decrease, at ALT (10.6 +/- 0.6 mmol x L(-1)) vs. SL (12.9 +/- 1.4 mmol x L( 1)), was only approximately 50% of that observed for IE (6.7 +/- 1.6 mmol x L(-1) vs. 11.3 +/- 2.8 mmol x L(-1)). Muscle power output and blood lactate accumulation during short supramaximal exercise are substantially unaffected by chronic hypoxia. PMID- 11472306 TI - Comparison of fatigue produced by various electrical stimulation trains. AB - Previous work has shown that variable-frequency trains (VFTs) that use an initial doublet to take advantage of the catch-like property of muscle produce more force in fatigued muscle than constant-frequency trains (CFTs); however, it is unclear whether repetitive activation with VFTs is more or less fatiguing than repetitive activation with CFTs. The purpose of this research was to investigate the forces and fatigue produced by various stimulation trains during repetitive isometric muscle contractions. Two force measurements, peak force and force-time integral, were used to measure the performance of the human quadriceps muscle. Three fatiguing protocols, each consisting exclusively of either CFTs, trains with an initial doublet (VFTs), or trains with doublets separated by longer intervals [doublet-frequency trains (DFTs)], were tested. In addition, force responses to each of the three train types were tested before and immediately following each fatiguing protocol. Regardless of the fatiguing protocol, the doublet-frequency testing trains produced the greatest peak forces and force-time integrals before and immediately following the fatiguing protocols. Repetitive activation with exclusively DFTs produced greater attenuation of the testing trains than repetitive activation with CFTs or VFTs. These results suggest that clinical applications of electrical stimulation to activate skeletal muscle may need to contain a combination of train types to optimize performance. PMID- 11472307 TI - Additive hypothermic effects of the 5-HT1A receptor agonist 8-OH-DPAT and the dopamine D2/3 receptor agonist 7-OH-DPAT in the rat. AB - The objective of this study was to examine possible interactions between serotonergic and dopaminergic agents lowering core temperature via stimulation of 5-HT1A and dopamine (DA) D2 receptors, respectively. The effects of the 5-HT1A receptor agonist (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin HBr (8-OH-DPAT) and the DA D2/3 receptor agonist 7-OH-DPAT on core temperature was monitored in adult male Wistar rats, approximately 300 g body weight. The temperature probe was connected to a PC-assisted temperature instrument, and an automated printer device was activated when the temperature reading had stabilized (+/-0.1 degrees C) for 10 s. As expected, 7-OH-DPAT [0.5 and 2.0 micromol x kg(-1) subcutaneous (s.c.)] as well as 8-OH-DPAT (0.15-2.4 micromol x kg(-1) s.c.), produced a dose dependent hypothermia. When combined, there were additive effects of the two compounds, although the effects of 7-OH-DPAT were attenuated by 8-OH-DPAT at the higher doses (0.6-2.4 micromol x kg(-1)), in all probability because of emerging DA D2 receptor blocking properties of the latter compound. PMID- 11472308 TI - Effects of heterotopic- and segmental counter-stimulation on the nociceptive withdrawal reflex in humans. AB - A nociceptive withdrawal reflex in 12 human volunteers was elicited by painful electrical stimulation applied to the cutaneous innervation area of the sural nerve. The evoked electromyographic reflex activities were recorded with surface electrodes placed on the short head of the biceps femoris muscle ipsi-lateral to sural nerve stimulation, before, during and after conditioning stimuli. The nociceptive withdrawal reflex was conditioned by nociceptive and non-nociceptive, heterotopic and segmental counter-stimulation. Heterotopic nociceptive counter stimulation and segmental nociceptive counter-stimulation were induced by ice water applied to the contra-lateral hand and foot, respectively. Heterotopic nociceptive counter-stimulation produced a significant inhibition of the nociceptive withdrawal reflex and the associated pains rating to sural nerve stimulation. Segmental nociceptive counter-stimulation inhibited the pain rating and tended to inhibit the nociceptive withdrawal reflex. There was no obvious correlation between visual analogue scale (VAS) and the nociceptive withdrawal reflex elicited by sural nerve stimulation neither in the group nor in the individual subject. This suggests that the nociceptive withdrawal reflex cannot always be used as a quantitative measure of pain. PMID- 11472309 TI - Changing dopaminergic activity through different pathways: consequences for renal sodium excretion, regional blood flow and oxygen tension in the rat. AB - Dopamine (DA) is an intrarenal natriuretic hormone involved in sodium homeostasis, but the regulation of renal dopaminergic tonus is unclear. We evaluated different pathways for elevating DA tonus to determine which are important for the ability of the kidney to produce natriuresis and studied the accompanying effects on regional renal blood flow and oxygen tension. Thus, we compared the effects of a catechol-O-methyl transferase (COMT)-inhibitor, an unspecific monoamine oxidase (MAO)-inhibitor, a D1-like receptor agonist and a DA precursor in anaesthetized rats. Sodium excretion increased sixfold after COMT inhibition, eightfold after administration of the D1-like agonist, whereas it was similar to control after MAO inhibition and infusion of DA precursor. Urinary dopamine excretion increased 42% by COMT inhibition, 55% by MAO inhibition and 12 fold after DA precursor, but remained unchanged after infusion of the D1-like agonist. The D1-like receptor agonist led to a 38% increase in the cortical blood flow and a 21% increase in outer medullary blood flow. Regional renal blood flow was unaffected by all other treatments. Cortical and outer medullary oxygen tension was unaffected in all treatment groups. To conclude, the natriuretic and haemodynamic properties of an elevation in DA tonus depends on the route by which the elevation occurred. Systemic administration of a D1-like receptor agonist, results in a natriuretic response which, as opposed to the natriuresis seen after COMT inhibition, coincides with an increase in renal cortical and outer medullary blood flow. Precursor delivery or MAO inhibition did not change neither urinary sodium excretion nor renal blood flow. PMID- 11472310 TI - Mitogen-activated protein kinase signal transduction in skeletal muscle: effects of exercise and muscle contraction. AB - Exercise has numerous growth and metabolic effects in skeletal muscle, including changes in glycogen metabolism, glucose and amino acid uptake, protein synthesis and gene transcription. However, the mechanism(s) by which exercise regulates intracellular signal transduction to the transcriptional machinery in the nucleus, thus modulating gene expression, is largely unknown. This review will provide insight on potential intracellular signalling mechanisms by which muscle contraction/exercise leads to changes in gene expression. Mitogen-activated protein kinase (MAPK) cascades are associated with increased transcriptional activity. The MAPK family members can be separated into distinct parallel pathways including the extracellular signal-regulated kinase (ERK) 1/2, the stress-activated protein kinase cascades (SAPK1/JNK and SAPK2/p38) and the extracellular signal-regulated kinase 5 (ERK5). Acute exercise elicits signal transduction via MAPK cascades in direct response to muscle contraction. Thus, MAPK pathways appear to be potential physiological mechanisms involved in the exercise-induced regulation of gene expression in skeletal muscle. PMID- 11472311 TI - Review article: a conceptual approach to understanding the molecular mechanisms of cancer development in Barrett's oesophagus. AB - Oesophageal adenocarcinoma is one of the most deadly human malignancies. Gastro oesophageal reflux disease (GERD) has been established as a strong risk factor for oesophageal adenocarcinoma, and more than 40% of adult Americans experience regular GERD symptoms. GERD can be complicated by oesophagitis, and by replacement of oesophageal squamous mucosa with metaplastic, intestinal-type epithelium (Barrett's oesophagus) that is predisposed to malignancy. Cancers in Barrett's oesophagus arise through a sequence of genetic alterations which endow unlimited proliferative capacity upon the cells by affecting components of the cell cycle clock apparatus-the pivotal molecular machinery in the cell nucleus that controls whether a cell will proliferate, differentiate, become quiescent or die. This report describes how the genetic abnormalities that have been recognized in Barrett's oesophagus might promote carcinogenesis through effects on the cell cycle clock machinery. The goal of this review is to provide the clinician with a useful conceptual basis for evaluating studies on the molecular mechanisms underlying the progression from metaplasia to carcinoma in Barrett's oesophagus. PMID- 11472312 TI - Review article: the risk of lymphoma associated with inflammatory bowel disease and immunosuppressive treatment. AB - Lymphoma complicating inflammatory bowel disease is well described. Whether the risk of lymphoma is increased by immunosuppressive treatment with azathioprine, 6 mercaptopurine or infliximab is a common concern among patients and physicians considering using these agents. This review aims to quantify the lymphoma risk in inflammatory bowel disease and the added risk attributable to these treatments. The evidence from published cases is that lymphomas occur at sites of active inflammatory bowel disease more often than expected for this to be a chance association. Studies on inflammatory bowel disease populations are conflicting, with some follow-up studies from large inflammatory bowel disease clinics showing an increase in lymphoma incidence, while other population-based studies show little or no increase in risk of lymphoma. A small increase in lymphoma risk in inflammatory bowel disease, perhaps 2-3-fold, may be compatible with both sets of data. Studies of the risks associated with immuno- suppression are less satisfactory, with smaller numbers of patients and relatively short follow-up. The available evidence would support a further increase in lymphoma risk associated with immunosuppressive treatment in inflammatory bowel disease of around fivefold compared to no immunosuppressive use, and tenfold compared to the general population. The risks appear to be less than that associated with renal and hepatic transplant-related immunosuppression. Infliximab treatment is still too new to make a full assessment of its long-term safety, but post-marketing surveillance currently suggests that lymphoma risk may not be any greater than that associated with azathioprine and 6-mercaptopurine. Population-wide surveillance for lymphoma in inflammatory bowel disease would be required to narrow the confidence intervals on these estimates of lymphoma risk in inflammatory bowel disease and immunosuppressive treatment. PMID- 11472313 TI - Review article: transfusion in gastrointestinal haemorrhage--if, when and how much? AB - Despite the conventional wisdom that blood transfusion is of use in acute upper gastrointestinal haemorrhage, this review proposes that there is considerable doubt about the optimal timing and intensity of transfusion of blood and crystalloids in this setting. It advocates the need for controlled trials to identify the most appropriate timing and intensity of transfusion of patients with acute gastrointestinal haemorrhage. PMID- 11472314 TI - Review article: lymphatic vessel pumping and inflammation--the role of spontaneous constrictions and underlying electrical pacemaker potentials. AB - The lymphatic circulation is important in maintaining tissue fluid homeostasis. It removes fluid, proteins and other particles from tissue spaces and returns them to the blood stream. This function is achieved by rhythmical contractions of the collecting lymphatic vessels. The contractile mechanism is intrinsic to the smooth muscles present in the vessel wall and consequent to action potentials. The underlying electrical mechanism has been proposed to be due to rhythmic synchronization of Ca2+-dependent spontaneous transient depolarizations. The lymphatic pumping activity adapts to changes in fluid load and has been observed to augment during inflammatory reactions to help resolve the associated oedema. This beneficial action has been generally attributed to the increase in interstitial pressure consequent to the oedema. However, little attention has been paid to the possible role inflammatory mediators that are present in the lymphatic vessel environment, could play in directly affecting the lymphatic contractile mechanism. This review article discusses our current knowledge on the mechanism and initiation of lymphatic pumping and how these events are modulated during inflammatory conditions. PMID- 11472315 TI - Renal tubular injury is present in acute inflammatory bowel disease prior to the introduction of drug therapy. AB - BACKGROUND: 5-aminosalicylic acid (5-ASA) has been associated with renal complications in inflammatory bowel disease. Renal function is typically monitored using serum creatinine; however, significant disease may predate increases in creatinine. AIMS: To identify whether markers of early renal disease (urinary albumin, alpha-1-microglobulin [alpha-1-M] and N-acetyl-beta-D glucosaminidase [NAG], and serum cystatin C) are useful in the assessment of renal function in inflammatory bowel disease patients receiving 5-ASA. METHODS: Twenty-one patients with a new diagnosis of inflammatory bowel disease were investigated. Samples were taken at diagnosis, and at 3-monthly intervals after the commencement of 5-ASA, for 1 year. RESULTS: Mean creatinine clearance was 100 mL/min and did not change following treatment. Inflammatory bowel disease was not associated with albuminuria. Urinary N-acetyl-beta-D-glucosaminidase and alpha-1 microglobulin at diagnosis were increased in 10 (48%) and 11 (52%) patients, respectively: treatment was not associated with consistent changes in urinary protein excretion. There was a significant correlation between cystatin C and creatinine clearance both at diagnosis (r=-0.533, P=0.0275) and combining the initial and follow-up data (r=-0.601, P < 0.01), but not between creatinine and creatinine clearance (P > 0.05). CONCLUSIONS: Tubular proteinuria is an extra intestinal manifestation of inflammatory bowel disease irrespective of 5-ASA treatment. Tubular proteins are not useful predictors of an adverse renal response to 5-ASA. Serum cystatin C may be an improved marker of glomerular filtration rate in this setting. PMID- 11472316 TI - A study of fructo oligosaccharides in the prevention of travellers' diarrhoea. AB - BACKGROUND: Prebiotic carbohydrates selectively stimulate the growth of bifidobacteria and lactobacilli in the human colon. These bacteria form part of the gut's inherent defence against invading pathogens. AIM: To test the effectiveness of fructo oligosaccharides in preventing travellers' diarrhoea. METHODS: A total of 244 healthy subjects, travelling to high and medium risk destinations for travellers' diarrhoea, took part in a randomized, double-blind, placebo-controlled study. The protocol comprised a preliminary week for recording bowel habit by diary, a 2-week pre-holiday period with the diary and consumption of 10 g of fructo oligosaccharides or placebo daily, followed by a 2-week holiday with continuation of treatment and diary. A post-study questionnaire was completed by all subjects on their return to the UK. RESULTS: The consumption of fructo oligosaccharides led to a small (6%; P < 0.02) increase in stool frequency in the pre-holiday period and gave a significantly better sense of 'well-being' during the holiday, although subjects reported more flatulence. There were non significant decreases in episodes of diarrhoea with 20% on placebo and 11% on fructo oligosaccharides recording episodes in the post-study questionnaire (P=0.08) and 46% placebo, 38% fructo oligosaccharides recording episodes in the diary (P > 0.1). No change in bowel frequency, consistency or stool size was recorded. CONCLUSION: Travel to high risk areas increases diarrhoea. Fructo oligosaccharides alone are not sufficient to prevent this, although do have some benefits for the subjects. PMID- 11472317 TI - Anal sphincter biofeedback and pelvic floor exercises for faecal incontinence in adults--a systematic review. AB - BACKGROUND: Faecal incontinence is a common health care problem. Biofeedback is extensively used in clinical practice to treat faecal incontinence. AIM: To systematically review and evaluate the evidence from clinical studies on the effectiveness of biofeedback as a treatment for faecal incontinence in adults. METHODS: A systematic literature search was undertaken using electronic databases, with review of the retrieved references. RESULTS: The search identified 46 studies published in English using biofeedback to treat adults complaining of faecal incontinence. Those studies included a total of 1364 patients. Of those studies with adequate data, 275 out of 566 patients (49%) were said to be cured of symptoms of faecal incontinence following biofeedback therapy and 617 out of 861 (72%) patients were reported to be cured or improved. Studies varied in the method of biofeedback used, criteria for success and the outcome measures used. Only eight of the 46 studies employed any form of control group. CONCLUSIONS: The data suggest that biofeedback and exercises help a majority of patients with faecal incontinence. However, methodological variation, lack of controls and a lack of validated outcome measures are problems in evaluating these results. PMID- 11472318 TI - Ambulatory gastric pH monitoring: proper probe placement and normal values. AB - BACKGROUND: Gastric acid production may persist while patients are treated with proton pump inhibitors. Twenty-four-hour intragastric pH monitoring is being used to identify gastric acid in the stomach while on medical therapy. AIM: To identify the optimal region of the stomach to demonstrate the presence of gastric acid. METHOD: Probe locations confirmed with fluoroscopy after placement and prior to removal. In experiment 1, five volunteers underwent simultaneous, 24-h gastro-oesophageal pH monitoring with the pH sensors located in the gastric antrum, body, fundus and distal oesophagus. In experiment 2, five volunteers underwent simultaneous 24-h pH monitoring with sensors located side by side in the gastric fundus assessing the reproducibility of gastric pH in this region. In experiment 3, 35 volunteers underwent 24-h pH monitoring with pH sensors located in the distal oesophagus and gastric fundus. The mean percentage time for which pH < 4 was calculated for total, upright, and supine time periods. RESULTS: pH profiles for the gastric fundus and body are similar-the mean percentage total time for which pH < 4 was 92.2% and 90.1%, respectively (P=N.S.). These values are significantly different from the antrum; pH < 4=54.6% (P < 0.01). pH values from the gastric fundus are highly reproducible (linear regression P= 0.004, r(2)=0.96). The normal values (mean +/- 95th percentile) for percentage time gastric pH < 4 in the fundus were: total 95.6 +/- 1.5%, upright 94.8 +/- 1.8%, and supine 96.5 +/- 2.3%. CONCLUSION: The fundus is the optimal location to evaluate the presence of gastric acid; pH values are highly reproducible in this area. Normal values for percentage time gastric pH < 4 for a healthy population are now defined. PMID- 11472319 TI - Double gastric infection with Helicobacter pylori and non-Helicobacter pylori bacteria during acid-suppressive therapy: increase of pro-inflammatory cytokines and development of atrophic gastritis. AB - BACKGROUND: Long-term acid suppression may accelerate the development of atrophic gastritis in Helicobacter pylori-positive subjects. The pathogenetic mechanism remains unclear. AIM: To test the hypothesis that gastric double infection with H. pylori and non-H. pylori bacterial species-during acid suppression-may result in an enhanced inflammatory response, contributing to the development of atrophic gastritis. PATIENTS AND METHODS: A consecutive series of patients with gastro oesophageal reflux disease undergoing treatment with proton pump inhibitors (n=113) or histamine2-receptor antagonists (H2-RAs) (n=37), and 76 non-treated dyspeptic controls were investigated. Gastric mucosal H. pylori and non-H. pylori bacteria, histological gastritis, H. pylori serology, and circulating interleukin (IL)-1beta, IL-6, and IL-8 were examined. RESULTS: Patients on acid suppression with either proton pump inhibitors or H2-RAs had a similar prevalence of H. pylori infection to the controls, but a higher prevalence of non-H. pylori bacteria (61% and 60% vs. 29%, P < 0.0001 and P < 0.002). Both the presence of H. pylori and non-H. pylori bacteria were independent risk factors of atrophic gastritis (antrum: relative risks (RRs), 10.1 and 5.07; corpus: RRs, 11.74 and 6.38). A simultaneous presence of H. pylori and non-H. pylori bacteria was associated with a markedly increased risk of atrophic gastritis (antrum: RR, 20.25; corpus: RR, 20.38), compatible with a synergistic effect. Furthermore, the simultaneous presence of both types of bacteria was associated with higher cytokine levels than in patients without any type of bacteria. This increase was also greater than in patients with H. pylori infection alone (P < 0.001, for both IL-1beta and IL-8). SUMMARY AND CONCLUSIONS: H. pylori-positive patients on long term acid inhibition displayed three features: non-H. pylori bacterial growth; increased cytokine levels; and a higher risk of atrophic gastritis. We suggest that double infection with H. pylori and non-H. pylori bacteria is a major factor in the development of atrophic gastritis during gastric acid inhibition. PMID- 11472320 TI - There are some benefits for eradicating Helicobacter pylori in patients with non ulcer dyspepsia. AB - BACKGROUND: The relationship between Helicobacter pylori infection and non-ulcer dyspepsia is not established. AIM: To determine whether eradication of H. pylori might be of benefit in non-ulcer dyspepsia patients. METHODS: We randomly assigned 129 H. pylori infected patients with severe epigastric pain, without gastro-oesophageal reflux symptoms, to receive twice daily treatment with 300 mg of ranitidine, 1000 mg of amoxicillin, and 500 mg of clarithromycin for 7 days and 124 such patients to receive identical-appearing placebos. RESULTS: Treatment was successful (decrease of symptoms at 12 months) in 62% of patients in the active-treatment group and in 60% of the placebo group (N.S.). At 12 months, the rate of eradication of H. pylori was 69% in the active-treatment group and 18% in the placebo group (P < 0.001). Complete relief of symptoms occurred significantly more frequently in patients on the active treatment (43%) than in placebo-treated patients (31%, P=0.048). Within the active-treatment group, therapeutic success was significantly more frequent in the non-infected patients (84% vs. 64%, P=0.04). CONCLUSIONS: Although eradicating H. pylori is not likely to relieve symptoms in the majority of patients with non-ulcer dyspepsia, a small proportion of H. pylori-infected patients may benefit from eradication treatment. PMID- 11472321 TI - Efficacy of ecabet sodium for Helicobacter pylori eradication triple therapy in comparison with a lansoprazole-based regimen. AB - BACKGROUND: The cytoprotective agent, ecabet sodium, inhibits urease activity and growth of Helicobacter pylori. AIM: To evaluate the efficacy and safety of ecabet sodium-based eradication of H. pylori infection, compared with a lansoprazole based regimen, in a randomized multicentre study. SUBJECTS AND METHODS: A total of 120 H. pylori-positive patients were assigned to one of two treatment regimens for 2 weeks: ecabet sodium 1 g b.d., amoxicillin 500 mg t.d.s. and clarithromycin 400 mg b.d. (EAC: 60 patients); or lansoprazole 30 mg (o.m.) with the same antimicrobial agents (LAC: 60 patients). Cure of infection was assessed by a 13C urea breath test 1 month after completion of treatment. RESULTS: One patient in the EAC group and two in the LAC group did not complete therapy because of an adverse event, and three did not undergo the 13C-urea breath test. Cure rates for the intention-to-treat, all-patients-treated and per protocol analysis in the EAC group were 85%, 86% and 88%, respectively, whereas those in the LAC group were 85%, 88% and 91%. There were no significant differences in cure rate or adverse events between the two regimens. CONCLUSIONS: Ecabet sodium in combination with amoxicillin and clarithromycin is as effective as lansoprazole-based eradication therapy for H. pylori. PMID- 11472322 TI - A triple therapy regimen after failed Helicobacter pylori treatments. AB - BACKGROUND: Following standard triple therapy, up to 20% of patients require further Helicobacter pylori eradication treatment. Data regarding the efficacy of re-treatment in these patients are scarce. AIM: To evaluate the efficacy of a triple therapy after one or more consecutive treatment failures. METHODS: A total of 51 patients with persistent H. pylori infection after at least one unsuccessful standard 1-week regimen were enrolled in the study. H. pylori infection at entry was assessed by rapid urease test and histology on biopsies from the antrum and the corpus. Patients were given a 2-week triple therapy, comprising ranitidine bismuth citrate 400 mg b.d., tetracycline 500 mg t.d.s., and tinidazole 500 mg b.d. Ranitidine bismuth citrate was given during meals, whilst tetracycline and tinidazole was given after meals. Bacterial eradication was assessed by endoscopy (36 patients) or 13C-urea breath test (15 patients) 4-6 weeks after therapy had ended. RESULTS: All 51 patients completed the study and H. pylori eradication was achieved in 46, with an eradication rate of 90% (95% CI: 82-98). In detail, bacterial eradication was obtained in 96% of patients who had previously failed one course of clarithromycin-amoxicillin based triple therapy, in 88% patients who had failed a clarithromycin-tinidazole based triple therapy, in 83% patients who had failed both treatment schedules, and in the only patient who had failed three consecutive therapeutic attempts. Two patients took the therapy for 9 and 10 days instead of the full 14 day-course. No major side effects were reported, whilst six (12%) patients complained of mild side-effects. CONCLUSION: This study demonstrates that this triple therapy regimen is effective for re-treatment of H. pylori infection. PMID- 11472323 TI - Ranitidine bismuth citrate with clarithromycin alone or with metronidazole for the eradication of Helicobacter pylori. AB - BACKGROUND: Both triple therapy with ranitidine bismuth citrate (RBC) plus two antibiotics for 7 days and dual therapy of RBC with clarithromycin for 14 days have been extensively studied; both regimens effectively eradicate Helicobacter pylori. However, few studies have assessed the efficacy of dual therapy given for 7 days. AIM: To compare the efficacy and safety of RBC 400 mg with clarithromycin 500 mg, alone or with metronidazole 400 mg, given twice daily for 7 days for the eradication of H. pylori. METHODS: This single centre, randomized, double-blind study involved 118 patients with dyspepsia or a history of peptic ulcer disease. H. pylori infection was detected initially by CLO test, and confirmed in 109 patients by urea breath test and/or microbiology culture. H. pylori eradication was assessed 4 and 12 weeks after the end of treatment by urea breath test. H. pylori antibiotic susceptibility was assessed pre-study in all patients, and post treatment in patients with a positive post-treatment urea breath test. Adverse events were recorded throughout the study. RESULTS: H. pylori was eradicated in 93% of patients who received RBC with clarithromycin and metronidazole and in 84% of patients who received RBC with clarithromycin (intention-to-treat rates). Per protocol eradication rates were 98% and 90% for triple therapy and dual therapy, respectively. The eradication of metronidazole-resistant H. pylori was achieved in 100% and 88% of patients following dual therapy and triple therapy, respectively, and acquired resistance to clarithromycin occurred in only one patient following treatment failure. Both treatments were well-tolerated; only one patient (2%) was withdrawn from each treatment group due to adverse events. CONCLUSIONS: RBC with clarithromycin and metronidazole is a highly effective and well-tolerated triple therapy regimen for the eradication of H. pylori. RBC with clarithromycin dual therapy has a similar efficacy, and offers an alternative to triple therapy when there are concerns about treatment with metronidazole or the use of multiple antibiotics. Both regimens are effective against antibiotic resistant strains of H. pylori. PMID- 11472324 TI - Helicobacter pylori testing in the primary care setting: which diagnostic test should be used? AB - AIM: To identify the most accurate and efficient test for diagnosing Helicobacter pylori infection in primary care patients. STUDY DESIGN: A whole blood test, an ELISA, and carbon13 urea breath test (CUBT) were evaluated in a primary care setting and validated against two different gold standards that used gastric biopsies. POPULATION: Primary care patients who had dyspeptic complaints lasting at least 2 weeks and were referred for endoscopy. OUTCOMES MEASURED: Positive and negative predictive values, sensitivity and specificity were determined for all three noninvasive H. pylori tests. RESULTS: Data from the three non-invasive H. pylori tests were available for 136 primary care dyspeptic patients referred for endoscopy. They were compared with data from the gold standards. The positive predictive value of the whole blood test was in the range 71-75%, the ELISA 83 86%, and the CUBT 88-92%, while the negative predictive values were in the ranges 72-77%, 96-100%, and 95-98%, respectively. The sensitivity of the whole blood test was in the range 36-42%, the ELISA 93-100%, and the CUBT 92-97%, while the specificities were in the ranges 92-93%, 90-91% and 93-95%, respectively. The positive predictive value of the ELISA dropped significantly at lower H. pylori infection rates. DISCUSSION: Both the ELISA and CUBT are effective in the primary care setting, while the whole blood tests produces inferior results. ELISA might, however, be less suitable for detecting H. pylori infection in a population with a low rate of infection. PMID- 11472325 TI - Glucocorticoids and IL-10, but not 6-MP, 5-ASA or sulfasalazine block endothelial expression of MAdCAM-1: implications for inflammatory bowel disease therapy. AB - BACKGROUND: Enhanced MAdCAM-1 (mucosal addressin cell adhesion molecule-1) expression is associated with the aetiology of inflammatory bowel disease, but little is known about MAdCAM-1: regulation, or how inflammatory bowel disease therapies modulate MAdCAM-1. AIM: To examine how agents currently used to treat inflammatory bowel disease affect MAdCAM-1: induced by tnf-alpha in an in vitro model of inflammatory bowel disease. METHODS: Endothelial monolayers were pretreated with dexamethasone (DEX): 5-aminosalicylic acid (5-ASA), 6 mercaptopurine (6-MP), sulfasalazine or interleukin-10: (IL-10: prior to TNF alpha (20 ng/mL), and MAdCAM-1: measured by Western blotting, RT-PCR, EMSA and lymphocyte adhesion assays. RESULTS: MAdCAM-1: was induced dose- and time dependently by TNF-alpha on endothelial cells. Either dexamethasone or IL-10: reduced TNF-alpha-induced MAdCAM-1: protein, mRNA and lymphocyte adhesion. However, neither 5-ASA, sulfasalazine nor 6-MP blocked MAdCAM-1 induction. CONCLUSIONS: Our data indicate that dexamethasone or IL-10 can exert therapeutic activity in inflammatory bowel disease through MAdCAM-1 inhibition. 5-ASA, sulfasalazine and 6-MP, while beneficial in inflammatory bowel disease, do not directly control MAdCAM-1, and are beneficial through inhibition of other inflammatory processes. PMID- 11472326 TI - Probiotic impact on microbial flora, inflammation and tumour development in IL-10 knockout mice. AB - BACKGROUND: The enteric bacterial flora has been implicated in the pathogenesis of enterocolitis and colon cancer in C57BL/6 IL-10 knockout mice. Probiotic Lactobacilli modify the enteric flora and are thought to have a beneficial effect on enterocolitis. We conducted a controlled feeding trial in IL-10 knockout mice using the probiotic Lactobacillus salivarius ssp. salivarius UCC118. AIM: To determine the effect of probiotic consumption on the gastrointestinal microflora, tumour development and colitis in IL-10 knockout mice. METHODS: Twenty IL-10 knockout mice were studied (10 consumed probiotic organisms in milk and 10 consumed unmodified milk) for 16 weeks. Faecal microbial analysis was performed weekly to enumerate excretion of the probiotic UCC118, total lactobacilli, Clostridium perfringens, bacteroides, coliforms, bifidobacteria and enterococci. At sacrifice, the small and large bowel were microbiologically and histologically assessed. RESULTS: L. salivarius UCC118 was detected in faeces from all mice in the probiotic fed group, but not the control group. Faecal coliform and enterococci levels were significantly reduced in probiotic fed animals compared to the controls (P < 0.05). At sacrifice, a significant reduction in C. perfringens numbers was observed in the test mice (P < 0.05). There were no fatalities in the test group compared to two deaths from fulminant colitis in the control group. Only one test mouse developed colonic adenocarcinoma compared to five in the control group. Test animal mucosal inflammation consistently scored lower than that of the control mice. CONCLUSION: In this placebo controlled trial, modification of enteric flora in IL-10 knockout mice by probiotic lactobacilli was associated with reduced prevalence of colon cancer and mucosal inflammatory activity. PMID- 11472327 TI - Interleukin-8 increases acetylcholine response of rat intestinal segments. AB - BACKGROUND: Interleukin-8 (IL-8) is a pro-inflammatory cytokine highly expressed in inflammatory bowel diseases, but whose effects on intestinal motility are unknown. AIM: To characterize the role of IL-8 in the contraction of rat intestinal segments. METHODS: Contractile response to acetylcholine (ACh 10-6 M) in terminal ileal segments (including mucosa) from Wistar rats was measured before and after incubation (15, 30, 60 or 90 min) with IL-8 (1 ng/mL), and after 60 min of incubation with different doses of IL-8 (0, 0.1, 0.5, 1, 10 and 100 ng/mL). The effects of blocking neural transmission with tetrodotoxin (TTX) and inhibiting protein synthesis (cycloheximide) were tested. The contractile response of longitudinal muscle-myenteric plexus preparations (i.e. without mucosa) was measured after 60 min of incubation with 0.1 and 1 ng/mL of IL-8. RESULTS: IL-8 increased ileal contraction induced by ACh 10(-6) M. This augmentation was significant after 60 min of incubation (58%, P=0.01) and persisted after 90 min (18%, P=0.04). A 60-min incubation period showed a dose related effect, beginning at 0.5 ng/mL (30%, P=0.003) and reaching a peak at 1 ng/mL (58%, P=0.01). The same effect was also observed on colonic segments. TTX did not affect the IL-8 increase of ACh-induced contractions, which was completely abolished by cycloheximide. IL-8 had no significant effect on longitudinal muscle-myenteric plexus preparations. CONCLUSION: In vitro, IL-8 increases contractile response of the ileum to ACh in a dose-dependent manner. This effect is not neurally mediated, but seems to involve protein synthesis by intestinal mucosa. PMID- 11472328 TI - Duodenogastric reflux following cholecystectomy in the dog: role of antroduodenal motor function. AB - BACKGROUND: Duodenogastric reflux has been implicated in the pathogenesis of gastric ulcer and gastritis. Duodenogastric reflux after cholecystectomy is also a possible cause of post-cholecystectomy syndrome. AIM: To investigate the role of antroduodenal motor function in increased duodenogastric reflux following cholecystectomy and the effect of trimebutine maleate (trimebutine) on the duodenogastric reflux in conscious dogs. METHODS: Antropyloric and duodenal motility and bile acids content in the gastric juice were measured for 3 h during the inter-digestive state in dogs with or without cholecystectomy. RESULTS: Bile acids content in the gastric juice of cholecystectomized dogs was significantly higher than that of non-cholecystectomized dogs. The frequency of pyloric relaxation during phase II of the migrating motor complex was significantly increased following cholecystectomy. Intravenous infusion of trimebutine inhibited both the increased duodenogastric reflux and the frequency of pyloric relaxation in the cholecystectomized dog. CONCLUSION: Duodenogastric reflux and frequency of pyloric relaxations were increased in cholecystectomized dogs and trimebutine suppressed both of them. These findings suggest that the increased frequency of pyloric relaxation contributes to the duodenogastric reflux following cholecystectomy. PMID- 11472329 TI - Reconstruction of seminal ducts in obstructive azoospermia. AB - Depending on the localization of the obstruction of the seminal ducts, either a microsurgical reconstruction (tubulovasostomy, vasovasostomy) or a transurethral resection of the ejaculatory ducts is carried out. We have compared the effectiveness and economic advantages of reconstructive microsurgery of the epididymis and vas deferens with standard procedures in animal experiments. Microsurgical invagination techniques in tubulovasostomy are equal to the standard procedure from the point of view of the patency and fertility rates. They are also easier to learn and carry out. Less time is required for the invagination technique, and also less microsurgical suture material. The double layer technique in vasovasostomy is equal to the one-layer microsurgical technique from the point of view of patency and fertility rates. The one-layer technique requires less time and suture material. It seems that the discrepancy between the patency and the fertility rate is related to immunological processes after reconstruction of the seminal ducts. In cases of obstructive azoospermia it is necessary to investigate the individual conditions and possibilities of the infertile couple. As a result of the high success rate obtainable today by surgical reconstruction of the seminal ducts, this must constitute the first type of treatment to be considered, before any of the procedures of reproductive medicine are undertaken. PMID- 11472330 TI - Mobility shift assay of calcium-binding proteins of mouse epididymal spermatozoa. AB - The calcium-binding proteins (CBPs) of mouse epididymal spermatozoa were analysed by mobility changes in the presence of added Ca2+ in two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The expression patterns of relatively high molecular weight CBPs (Mr > 20 kDa) were different between caput and cauda epididymal spermatozoa. There was a constitutive expression of low molecular weight CBPs (Mr < 20 kDa) regardless of the epididymal region. Most of the CBPs disappeared after the acrosome reaction (AR) induced by Ca2+ ionophore A23187, suggesting that they originated from the acrosome and/or the plasma membrane overlaying the acrosome. Taken together, it can be suggested that changes in CBPs of spermatozoa are important features of sperm maturation during epididymal transit, and that they may be related to the fertilizing ability of mouse epididymal spermatozoa. PMID- 11472331 TI - Differential binding of X- and Y-chromosome-bearing human spermatozoa to zona pellucida in vitro. AB - The sex of human offspring has been associated with the day in the mother's menstrual cycle on which insemination occurs, with male zygotes being formed earlier in the fertile period than female zygotes. Using an in vitro environment designed to mimic the in vivo milieu, we tested the hypothesis that Y-chromosome bearing spermatozoa survive functionally longer than X-chromosome-bearing spermatozoa, and that this differential functional survival is a contributing factor to the in vivo phenomenon. Donor semen was processed by swim-up and incubated at 37 degrees C in culture medium for 0, 24 and 48 h, with human zona pellucida (hemizona, HZ) being used to select functional spermatozoa. A second set of in vitro storage conditions, 4 degrees C in test-yolk refrigeration buffer, was used to determine whether changing the incubation conditions alters the process. The sex chromosome of the spermatozoa was determined using fluorescent in situ hybridization (FISH). For spermatozoa incubated at 37 degrees C, the percentage of functional (HZ bound) Y-bearing spermatozoa was significantly increased (P < 0.05) at 48 h (55.4 + 2.9%) but not at 0 h (50.5 + 0.7%) or 24 h (52.8 + 3.1%) compared to swim-up spermatozoa (50.6 + 0.3%). No difference in the percentage of functional Y-spermatozoa was observed at any time point with storage at 4 degrees C in refrigeration buffer. Thus, we demonstrated that significantly more Y-bearing spermatozoa were capable of zona binding than X bearing spermatozoa at 48 h at 37 degrees C incubation, with an observed Y : X ratio of 1.15 for these zona-bound spermatozoa compared to 1.02 for post-swim-up spermatozoa. We conclude that a differential functional survival appears to exist between X-bearing and Y-bearing spermatozoa, with the latter exhibiting a longer functional survival under in vitro conditions. PMID- 11472332 TI - Evaluation of the effect of a cervical cap device on sperm functional characteristics in vitro. AB - Intracervical insemination continues to be employed for homologous and donor insemination in natural and stimulated cycles. Efficacy studies for potential fertility involve in vivo assessment; however, in vitro testing of particular sperm function(s) critically involved in fertilization is an important component of such evaluation. We report here on the in vitro evaluation of the effects of the silicone Veos cervical cap (Veos, London, UK) on sperm function. Donor semen was exposed to the Veos cervical cap or a sterile 15-cc centrifuge tube (control), or treated with the spermicide nonoxynol-9 (5 mg x ml(-1) in saline) for 4 h at 37 degrees C and 5% CO2 in water-saturated air. After exposure, motility characteristics, both in semen and in spermatozoa processed by standard swim-up procedure, cervical mucus penetration and sperm-zona pellucida interaction using the hemizona assay were assessed. Results indicated that exposure to the Veos cervical cap had no effect on either sperm motility characteristics or sperm-zona pellucida interaction. A small but significant difference was observed for cervical mucus penetration (P = 0.05); however, for both the control and treated groups, vanguard spermatozoa exceeded manufacturer's guidelines for a normal test, a penetration distance of > or = 30 mm. As expected, nonoxynol-9 was a potent inhibitor of sperm function. Lack of adverse effects on in vitro spermatozoa functional characteristics after exposure to the silicone Veos cervical cap supports its addition to the repertoire of fertility treatment modalities when cervical insemination is indicated. PMID- 11472333 TI - Development of a new, highly sensitive zona pellucida binding assay using a bioluminescence-enhanced detection system. AB - To date, two different zona binding assays have been described in the literature. Both assays, however, require a large quantity of human zonae which vary immensely in quality. Furthermore, an inverted microscope with micromanipulation equipment is necessary, which makes both assays relatively complicated and time consuming, and requires skilled staff. Therefore, we developed a new, highly sensitive zona binding assay using a bioluminescence-enhanced system which employs a pool of solubilized zona pellucida and is easier for routine use. In the detection system, light emission by the luciferin-luciferase system is measured. Because of the limited availability of human zonae pellucidae, this new assay was first developed in the porcine system. The new bioluminogenic substrate D-luciferin-O-beta-galactopyranoside (Lu-Gal) was synthesized, purified and characterized. Synthesis of Lu-Gal resulted in purity better than 99.998%. Analytical data and spectra were appropriate. In terms of the kinetic data, Lu Gal is a highly sensitive and specific substrate for beta-galactosidase. Using the given chemical conditions, nonlabelled zonae bound competitively to boar spermatozoa, which resulted in a high sensitivity and specificity. By the addition of 10 nonlabelled zonae, the binding of labelled zonae was almost completely inhibited. Corresponding results were obtained when the bioluminescent system was compared with the hemizona assay. On the other hand, spermatozoa of other species (bull, hamster and man) showed only low binding to the porcine zonae or none at all. Competitive displacement was not observed, indicating the inter-species specificity of the assay. PMID- 11472334 TI - An improved method of sperm selection by glass wool filtration. AB - An improved method of sperm selection by glass wool filtration is introduced. After incubation of glass wool filtrates for 30 min at 37 degrees C in a conical shaped 1.5-ml tube, an enrichment of highly motile spermatozoa was found in the bottom layer of the tube. The effect turned out to be dependent on the conical shape of the tube, as it was not observed in flat-bottomed tubes. Native ejaculates (obtained from 30 men) and their glass wool filtrates were analysed by cell counter, computer-assisted sperm-motility analysis, morphological differentiation and supravital staining of spermatozoa. When 400 microl of ejaculate, diluted with 800 microl of medium, was applied to the top of a column consisting of a 1-ml disposable syringe barrel gently packed with 15 mg of glass wool to a depth of 6 mm, an enrichment of viable spermatozoa was found in the first three 100-microl fractions taken from the bottom of the tube. It is the simplicity of this technique that makes it so easily applicable. PMID- 11472336 TI - Normal fetal bone. PMID- 11472337 TI - Hepatosplenic T-cell lymphoma. PMID- 11472338 TI - Congenital alpha(2)-plasmin inhibitor deficiencies: a review. PMID- 11472339 TI - Non-MHC-restricted cytotoxic cells: their roles in the control and treatment of leukaemias. PMID- 11472340 TI - Does red-cell T activation matter? PMID- 11472341 TI - Relationship between graft-versus-host disease and graft-versus-leukaemia in partial T cell-depleted bone marrow transplantation. AB - The success of allogeneic bone marrow transplantation (BMT) is limited by the major complications, graft-versus-host disease (GVHD) and relapse. The very beneficial effect of maximal T-cell depletion of the graft for prevention of GVHD has been counterbalanced by an increase in graft failure and relapse of disease. Therefore, we started an approach of partial T-cell depletion of the graft. GVHD and graft-versus-leukaemia (GVL) are strongly correlated after non-T cell depleted BMT. Here, we report whether the correlation between GVHD and GVL also exists in partial T cell-depleted BMT from sibling donors. We retrospectively studied 117 adult patients with early haematological malignancies. Our method of partial T-cell depletion gave a relapse rate in patients with acute leukaemias similar to that observed in non-T cell-depleted BMT. However, patients with chronic myeloid leukaemia had a relapse rate that was similar to that observed in maximal T cell-depleted BMT. We found a significant correlation between the presence of chronic GVHD and an improved disease-free survival. Nevertheless, overall survival was lower in patients with chronic GVHD. There was no correlation between the occurrence of acute GVHD and disease-free or overall survival. PMID- 11472342 TI - HLA-A3 increases and HLA-DR1 decreases the risk of acute graft-versus-host disease after HLA-matched sibling bone marrow transplantation for chronic myelogenous leukaemia. AB - Frequencies of human leucocyte antigens (HLA)-A, -B and -DR were determined in 751 patients with chronic myelogenous leukaemia (CML) reported to the European Group for Blood and Marrow Transplantation after bone marrow transplantation from HLA-identical family donors and related to the occurrence of graft-versus-host disease (GVHD). HLA-A3 and DR1 were significantly associated with acute GVHD, the first with a higher risk (44% in HLA-A3(+) versus 34% in HLA-A3(-) patients) and the latter with a lower risk (28% in HLA-DR1(+) versus 38% in HLA-DR1(-) patients) for developing acute GVHD grade II--IV. Both factors were independent of known variables for GVHD as shown in a multivariate analysis. The results show that MHC alleles independently influence the incidence of GVHD in bone marrow transplantation from an HLA-identical donor for first chronic-phase CML. Possible mechanisms might include an HLA antigen-specific allele-associated effect, and/or non-specific allele-associated immune hypo- or hyper-responsiveness. PMID- 11472343 TI - Factors affecting lymphocyte subset reconstitution after either related or unrelated cord blood transplantation in children -- a Eurocord analysis. AB - Immune recovery after cord blood transplantation (CBT) is of concern owing to the low number of lymphocytes transferred with the graft and their immaturity. Risk factors influencing lymphocyte subset reconstitution related to disease, patient, donor and transplant were studied in 63 children (< 16 years), given either related (n = 14) or unrelated (n = 49) CBT for malignant (n = 33) or non malignant diseases (n = 30). Only children with sustained myeloid engraftment were analysed. Absolute numbers of T (CD3(+), CD4(+), CD8(+)), B and natural killer (NK) cells were reported 2--3, 6, 9, 12 and 12--24 months after CBT. Median patient age was 4.0 years (0--15) and median follow-up was 23 months (1.7- 61.0). Twenty-six patients received human leucocyte antigen (HLA)-matched CBT and 37 received HLA-mismatched CBT. The median number of nucleated cells (NCs) collected/recipient weight was 6.1 x 10(7)/kg. In this selected population, the estimate 2 year survival was 85%. Lymphocyte reconstitution (defined as the median time to reach the normal value of age-matched healthy children) was 3, 6 and 8 months for NK, B and CD8(+) cells, while it was 11.7 months for both CD3(+) and CD4(+) lymphocytes. In the multivariate analysis, factors favouring T-cell recovery were: related donor (P = 0.002); higher NCs/kg (P = 0.005) and recipient cytomegalovirus (CMV)-positive serology (P = 0.04). Presence of acute graft versus-host disease (GVHD) delayed T-cell recovery (P = 0.04). To summarize, in children with sustained myeloid engraftment the concern that lymphocyte recovery after CBT could be delayed does not appear to be substantiated by our results. PMID- 11472344 TI - In vivo expansion of the endogenous B-cell compartment stimulated by radiation and serial bone marrow transplantation induces B-cell leukaemia in mice. AB - Chronic lymphocytic leukaemia (CLL) is a malignancy of CD5(+) B cells. This B cell lineage is established during ontogeny and replenished by the process of self-renewal. Spontaneous and induced leukaemias that frequently affect this lineage are thought to arise as a result of the frequent cell division required to maintain the population throughout adulthood and in response to repeated exposure to environmental antigens. In a series of bone marrow transplant (BMT) experiments performed in B6D2F1 mice, B-cell leukaemia occurred in recipients of serially transplanted syngeneic bone marrow. This study was therefore designed to determine the frequency and phenotype of the observed leukaemia. Male donor cells were initially transplanted into lethally irradiated female hosts and secondary (2 degrees ) BMT was performed at 3 months. At 1, 2, 3 and 16 months following primary (1 degrees ) BMT, and when 2 degrees BMT recipients developed leukaemia, animals were sacrificed and their tissues extensively examined. These analyses confirmed a host-derived CD5(+) transplantable B-cell leukaemia that was initiated in 50% of 1 degrees BMT recipients. With serial passage, the leukaemia became more aggressive and lost CD5 expression, suggesting transformation to a high-grade leukaemia/lymphoma. This previously unreported observation suggests that the combination of radiation and subsequent serial transplantation induces a proliferative stress to the host B-cell compartment that is causative in leukaemic transformation. PMID- 11472345 TI - In vitro T-cell receptor V beta repertoire analysis may identify which T-cell V beta families mediate graft-versus-leukaemia and graft-versus-host responses after human leucocyte antigen-matched sibling stem cell transplantation. AB - We studied oligoclonal T-cell expansions of 24 T-cell receptor (TCR) V beta families in normal donor lymphocytes stimulated with patient's cells and in recipient blood after transplant, using a polymerase chain reaction-based assay (spectratyping). T cells from donor blood were incubated with separated myeloid leukaemia cells or T cells from the HLA-identical sibling recipient. In five of the six patients tested, the T-cell V beta skewing pattern observed in vitro was seen in vivo after transplant. After transplant, the myeloid-specific V beta skewing coincided with the disappearance of residual disease in three patients and in one patient skewing was lost at the time of leukaemic relapse. In functional tests, T cells generated against leukaemic cells in vitro produced interferon gamma in response to the leukaemia. Removal of the leukaemia-expanded skewed V beta families significantly decreased cytotoxic killing of the leukaemia. However, while there was a general concordance in the V beta family exhibiting clonal expansion in vitro and in vivo, the exact clonotype expanded in vitro and in vivo differed. These findings suggest that alloresponses involve multiple T-cell clones within a restricted TCR V beta repertoire that undergo different selection pressures in vitro and in vivo. PMID- 11472346 TI - Aberrant expression of caspase cascade regulatory genes in adult T-cell leukaemia: survivin is an important determinant for prognosis. AB - Derangement of either apoptosis or cell division is known to play an important role in tumorigenesis. Fas-mediated apoptosis on normal and leukaemic T cells is finely tuned by inhibitory proteins, such as FAP-1, FLIP and survivin, and defective caspase isoform which can attenuate the function of its intact caspase as a decoy molecule. However, complex involvement of such inhibitors in tumour biology relating to apoptotic pathology remains unclear in the neoplasms. We report the aberrant expression of FAP-1, FLIP and survivin mRNAs on leukaemic T cells from adult T-cell leukaemia (ATL) patients. Among these inhibitors, only survivin was aberrantly expressed in all ATL cases, but not in any normal peripheral blood mononuclear cells (PBMCs). Furthermore, survivin mRNA expression level was characteristic in each subtype of ATL and represented an important determinant for ATL prognosis. However, the apoptotic effector of casp-8, which is essential in Fas-mediated signal transduction, was dominant in defective casp 8 rather than intact casp-8 in ATL cells, suggesting a favourable biological situation for escape from apoptosis. Taken together, ATL cells probably possess many different regulatory mechanisms in order to attenuate Fas-mediated signalling and subsequently expand their populations under escape from apoptosis. Among these inhibitors, survivin is a useful bio-marker to assess tumour biology and may be a potential new target for apoptosis-based selective therapy in neoplasms as the expression is a general feature of neoplasia, but not normal tissues. PMID- 11472347 TI - Flavopiridol circumvents Bcl-2 family mediated inhibition of apoptosis and drug resistance in B-cell chronic lymphocytic leukaemia. AB - Flavopiridol, a synthetic flavone, is currently under clinical investigation for the treatment of B-cell chronic lymphocytic leukaemia (B-CLL). In this study, we examined the in vitro effects of flavopiridol and fludarabine on B-CLL cells from 64 patients (36 treated and 28 untreated) in terms of apoptosis induction and Bcl 2 family expression. Both flavopiridol and fludarabine induced apoptosis in all the samples tested with mean LD(50) values (+/- SD) of 59.7 nmol/l (+/- 36.5) and 6.2 micromol/l (+/- 7.5) respectively. Mean flavopiridol LD(50) values were not significantly different between the treated and untreated patient groups (P = 0.35), whereas the fludarabine LD(50) values were significantly higher in the previously treated patient group (P = 0.01). Bcl-2 and Mcl-1 expression were downregulated in both flavopiridol and fludarabine-induced apoptotic cells, but the increase in Bax expression that accompanied fludarabine-induced apoptosis was not evident in flavopiridol-treated cells. In addition, Bcl-2:Bax ratios were not predictive of flavopiridol cytotoxicity (P = 0.82), whereas they were highly predictive of in vitro responsiveness to fludarabine (P = 0.001). Overall, these findings suggest that flavopiridol exerts its cytotoxic effect through a novel cell-death pathway that is not subject to the Bcl-2 family mediated resistance mechanisms that reduce the efficacy of many conventional chemotherapeutic drugs. PMID- 11472348 TI - Safety and efficacy of thalidomide in patients with myelofibrosis with myeloid metaplasia. AB - We administered the anti-angiogenic drug thalidomide to 21 patients (12 men) with myelofibrosis with myeloid metaplasia (MMM), who were not responsive to standard treatment. Patients received thalidomide at an escalating dose from 100 to 400 mg/d. Administration of the drug was discontinued before the planned 6 months of treatment in 19 patients (90.5%), mainly because of somnolence and/or fatigue, neurological symptoms or neutropenia. Of the 13 evaluable patients (who received more than 30 d of therapy), anaemia improved in three out of seven (43%) who were treated because of anaemia; thrombocytopenia improved in two out of three (66.6%) who were treated because of thrombocytopenia; splenomegaly was reduced in four (30.8%). Undesired increases in white blood cell and platelet counts were observed in three (23.1%) and five (38.5%) patients respectively. A severity score, indexed on haematological and clinical parameters, improved in two patients (15.4%), but worsened in five (38.5%). In conclusion, standard-dose thalidomide in MMM patients is burdened with a high rate of side-effects, which prevent prolonged treatment. Because the drug is effective in improving anaemia and thrombocytopenia and in reducing splenomegaly, low-dose therapy warrants evaluation. The unexpected observation of leucocytosis and thrombocytosis suggests biological studies and better criteria for selection of patients for treatment. PMID- 11472349 TI - Antisense oligodeoxynucleotides to latent membrane protein 1 induce growth inhibition, apoptosis and Bcl-2 suppression in Epstein-Barr virus (EBV) transformed B-lymphoblastoid cells, but not in EBV-positive natural killer cell lymphoma cells. AB - Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP-1) is essential for immortalization of B cells by EBV, protects the infected cells from apoptotic cell death and induces Bcl-2 expression. Suppression of LMP-1 expression by antisense oligodeoxynucleotides (AS-oligo) to LMP-1 inhibits proliferation, promotes apoptosis and suppresses Bcl-2 expression in EBV-transformed B cells. However, the function of LMP-1 expression in EBV-positive natural killer (NK) cell lymphoma cells has not been reported previously. We examined the function of LMP-1 in two EBV-positive NK cell lymphoma cell lines (NK-YS and YT) through suppressing LMP-1 expression by AS-oligo to LMP-1. The AS-oligo to LMP-1 suppressed LMP-1 mRNA and protein expression in two EBV-positive NK cell lymphoma cell lines, as well as in an EBV-transformed B-cell line (CMG-1). Proliferation was inhibited, apoptosis was induced and Bcl-2 expression was suppressed in CMG-1 cells, but none of these events were observed in NK-YS or YT cells. These results suggest that proliferation, inhibition of apoptosis and Bcl-2 expression in EBV positive NK cell lymphoma cells are not directly regulated by LMP-1 as in EBV transformed B-cell lines, but are probably mediated through other signal transducing systems. PMID- 11472350 TI - Clinical significance of Aspergillus fungaemia in patients with haematological malignancies and invasive aspergillosis. AB - The clinical significance of Aspergillus fungaemia in the setting of a deep seated aspergillosis has not been clearly established. Among 107 microbiologically documented Aspergillus infections in patients with haematological diseases observed over a 17-year period, blood cultures grew Aspergillus species from 10 cases. Aspergillus fungaemia was documented in 9 out of 89 (10.1%) patients with pulmonary aspergillosis at a median of 5 d from the onset of clinical signs of infection, and in one patient with central venous catheter focal infection. Five (50%) patients died as a result of fungal infection a median of 12 d (range 4--48) from the documentation of Aspergillus fungaemia. A comparison between cases of invasive aspergillosis with or without fungaemia showed that fungaemic patients were similar to those without positive blood cultures regarding clinical presentation, risk factors, clinical course and outcome. The diagnostic role of Aspergillus fungaemia in the setting of a deep seated infection is limited because blood cultures become positive when a microbiological or clinical diagnosis of aspergillosis has already been performed. Aspergillus fungaemia does not necessarily seem to be correlated with a disseminated infection or a poorer prognosis. PMID- 11472351 TI - Pretreatment characteristics and clinical outcome of acute promyelocytic leukaemia patients according to the PML-RAR alpha isoforms: a study of the PETHEMA group. AB - Of 167 newly diagnosed acute promyelocytic leukaemia patients, 83 patients were long (L)-form (50%), eight variable (V)-form (5%) and 76 short (S)-form (45%). The V-form and S-form groups presented a significantly higher percentage of patients with white blood cell counts > 10 x 10(9)/l (P < 0.05). The S-form cases displayed a significantly higher number of cases with M3v microgranular features (P = 0.005) and CD34 expression (P < 0.0001). There were no differences between the three isoforms in complete remission (CR) rate (overall CR 90%), but the 3 year disease-free survival was lower for V-form cases than it was for L- and S form cases (62% vs. 94% and 89%, P = 0.056). We conclude that the V-form and S form types are associated with some negative prognostic features at diagnosis. However, our data were only able to demonstrate an association with adverse prognosis in the V-form type and, moreover, as the number of cases was limited, needs to be confirmed in large, uniformly treated series. PMID- 11472352 TI - Clearance of maternal leukaemic cells in a neonate. AB - A 36-week pregnant woman was diagnosed with acute lymphoblastic leukaemia. Delivery was initiated prematurely, and a healthy child was born. Cord blood and peripheral blood samples from the neonate (obtained at 6 weeks, 3 months and 6 months) were analysed for the presence of minimal residual disease by polymerase chain reaction analysis of a leukaemia-specific IGH gene rearrangement and the E2A--PBX1 fusion gene transcript. In the cord blood sample, a tumour load of approximately 4 x 10(-4) was found, whereas all later blood samples were negative. Our data indicate that the maternal leukaemic cells did not engraft in the neonate. PMID- 11472353 TI - Response to vaccination against different types of antigens in patients with chronic lymphocytic leukaemia. AB - We investigated responses to vaccination against pneumococcal polysaccharide, Haemophilus influenzae b (Hib) conjugate and tetanus toxoid antigens in 31 patients with chronic lymphocytic leukaemia (CLL) and 25 controls. While in the control group all antibody responses against different antigens were highly significant, in the patient group clear evidence for responsiveness was detected only in the case of Hib polysaccharide antigen. Certain CLL patient subgroups showed low reactivity against tetanus toxoid antigen. In conclusion, plain polysaccharide vaccines seem to be ineffective in patients with CLL. Conjugate vaccines, in turn, are immunogenic and may offer protection against infections caused by encapsulated bacteria in these patients. Further studies concerning an optimal vaccination scheme and clinical efficiency are warranted. PMID- 11472354 TI - A phase II trial of pirfenidone (5-methyl-1-phenyl-2-[1H]-pyridone), a novel anti fibrosing agent, in myelofibrosis with myeloid metaplasia. AB - The anti-fibrotic and cytokine modulatory properties of pirfenidone suggest its usefulness in the treatment of myelofibrosis with myeloid metaplasia (MMM). In a prospective study, 28 patients with MMM were treated with oral pirfenidone. Twelve patients completed 1 year of therapy; 13 were withdrawn because of disease progression and three because of drug intolerance. Only one patient experienced a clinically relevant benefit with respect to anaemia and splenomegaly. The overall lack of clinical benefit correlated with no significant improvement in the bone marrow morphological features of the disease. We conclude that pirfenidone has no significant clinical or biological activity in MMM. PMID- 11472355 TI - High-dose factor VIIa increases initial thrombin generation and mediates faster platelet activation in thrombocytopenia-like conditions in a cell-based model system. AB - Clinical experience has shown that high doses of recombinant factor VIIa (rFVIIa) may ensure haemostasis in thrombocytopenic patients. We have used a cell-based model system to mimic thrombocytopenia and analyse the effect of rFVIIa. Lowering the platelet density from 200 x 10(9)/l (reflecting normal conditions) to 100, 50, 20 and 10 x 10(9)/l revealed a platelet density-dependent decrease in the maximal rate of thrombin generation, a prolongation in the time to maximal thrombin activity and a lower maximal level of thrombin formed. The platelet activation, measured as the time to half-maximal P-selectin (CD62) exposure, was not significantly dependent on the platelet density in the range of 200 x 10(9)/l to 10 x 10(9)/l, although there was a tendency for slower platelet activation at 20 x 10(9) and 10 x 10(9) platelets/l than at the higher platelet densities. Addition of 50--500 nmol/l rFVIIa to samples with 20 x 10(9) or 10 x 10(9) platelets/l shortened the lag phase of thrombin generation as well as the time to half-maximal platelet activation. Our data indicate that high doses of rFVIIa may help to provide haemostasis in thrombocytopenic patients by increasing the initial thrombin generation, resulting in faster platelet activation and thereby compensating for the lower number of platelets present. PMID- 11472356 TI - Cyclosporin A for the treatment of patients with chronic idiopathic thrombocytopenic purpura refractory to corticosteroids or splenectomy. AB - Patients with chronic immune thrombocytopenic purpura (ITP) who are unresponsive to corticosteroids require splenectomy, but if this fails, treatment is difficult. We tried to induce durable remissions in ITP patients refractory to corticosteroids before or after splenectomy by applying strong immunosuppression with the combination of cyclosporin A (CyA 5 mg/kg/d) and prednisone (0.4 mg/kg/d). Patients were assigned to one of two groups. Group 1, 10 patients refractory to prednisone; and group 2, 10 patients refractory to at least prednisone and splenectomy. Overall response rate was 55% (50% in group 1 and 60% in group 2 patients). Nine of the 10 patients in group 1 finally had a splenectomy because of relapse, insufficient response or toxicity of CyA. Thirty percent of the patients discontinued CyA because of side-effects; hypertension, severe headache and muscle pain being the most frequent encountered. It is concluded that CyA treatment does not avoid, but only postpones, splenectomy in chronic ITP patients who are refractory to corticosteroids. However, CyA can be useful in a subgroup of patients with corticosteroid- and splenectomy-refractory ITP, but treatment toxicity is high. PMID- 11472357 TI - The role of thrombopoietin in post-operative thrombocytosis. AB - Thrombopoietin (Tpo), the main regulator of thrombocytopoiesis, is a probable candidate to play a role in the increase in platelet counts that is frequently seen after surgery. In the current study, serial blood samples of patients that underwent major surgery were analysed with respect to Tpo kinetics, platelet turnover and inflammatory cytokines. Platelet Tpo content and plasma Tpo levels rose before platelet counts increased, suggesting that Tpo was indeed responsible for the elevation in platelet counts. In addition, an increase in interleukin 6 (IL-6) levels, but not in IL-11 and tumour necrosis factor alpha levels, was seen before the rise in Tpo concentration. In vitro, IL-6 was shown to enhance Tpo production by the HepG2 liver cell line. Thus, increased Tpo levels after surgery, possibly resulting from enhanced Tpo production under the influence of IL-6 or other inflammatory cytokines, are involved in an enhanced thrombocytopoiesis. PMID- 11472358 TI - RAFTK/Pyk2 involvement in platelet activation is mediated by phosphoinositide 3 kinase. AB - Platelet activation by different agonists initiates a signalling cascade involving the phosphorylation of several protein kinases, which control key regulatory events. Previously, we demonstrated that the related adhesion focal tyrosine kinase (RAFTK, Pyk2) was involved in an early phase of platelet activation, independent of integrin and glycoprotein IIb-IIIa activation. In this study, we demonstrate that RAFTK is co-immunoprecipitated with phosphoinositide 3 kinase (PI3K) upon platelet activation, and that thrombin, ADP and collagen induced the phosphorylation of both PI3K and RAFTK. A low dose of thrombin (0.015 U/ml) induced RAFTK phosphorylation and platelet aggregation in a PI3K activity dependent manner, whereas a high dose of thrombin (0.1 U/ml) induced these events in a PI3K activity-independent manner. ADP and collagen also induced RAFTK phosphorylation and platelet aggregation in a PI3K activity-dependent manner, similar to that of the low-dose thrombin. Furthermore, protein tyrosine phosphatase activity was associated with RAFTK in response to platelet activation, and was found to be that of protein tyrosine phosphatase-2 (SHP-2). The association of SHP-2 with RAFTK was PI3K-dependent and was increased upon RAFTK phosphorylation. Taken together, our results strongly suggest that the involvement of RAFTK in platelet activation is mediated via the PI3K pathway. PMID- 11472359 TI - Bruton's tyrosine kinase is present in normal platelets and its absence identifies patients with X-linked agammaglobulinaemia and carrier females. AB - X-linked agammaglobulinaemia (XLA) is a primary immunodeficiency caused by mutations in the gene coding for Bruton's tyrosine kinase (Btk) and is characterized by an arrest of B-cell development. We analysed Btk protein expression in platelets using flow cytometry and found that normal platelets express large amounts of Btk. Assessment of affected males from 45 unrelated XLA families revealed that platelets of the majority of the patients (37 out of 45 families) had decreased or absent Btk expression, and that platelets from carrier females of these families had both normal and mutated Btk expression, indicating that megakaryocytes in XLA carriers undergo random X-chromosome inactivation. These observations demonstrate that Btk is not crucial for maturation of megakaryocytes and the production of platelets. No correlation between Btk expression in platelets and clinical phenotype was observed in this study. Flow cytometric evaluation using platelets is a simple and rapid method to test Btk expression. It may be used as a screening test for XLA and for carrier detection, followed, if necessary, by more expensive mutation analyses. PMID- 11472360 TI - The C807T/G873A polymorphism in the platelet glycoprotein Ia gene and the risk of acute coronary syndrome in the Italian population. AB - Membrane glycoprotein (GP) Ia/IIa mediates platelet adhesion to collagen. The linked C807T/G873A polymorphisms in the GP Ia gene are correlated with a variable expression of the platelet surface receptor, the 807 TT/873 AA genotype being associated with a higher receptor density. Our study aimed to evaluate the possible role of the GP Ia C807T/G873A polymorphism as a risk factor for acute coronary syndrome in the Italian population. We investigated 157 patients with acute coronary syndrome (117 with myocardial infarction and 40 with severe unstable angina) as the first manifestation of coronary disease occurring before 65 years of age, compared with 312 healthy controls. All individuals were of Italian ancestry and were genotyped for the GP Ia C807T/G873A polymorphism. Complete linkage between the 807 and 873 sites was found in all samples. The 807 TT genotype was present in 12.7% of cases and in 4.8% of controls; the odds ratio for acute coronary syndrome was 2.9 (95% CI 1.4--5.8) for the 807 TT genotype compared with C-allele carriers and 0.6 (95% CI 0.4--0.9) for the 807 CC genotype compared with T-allele carriers. For the TT genotype, compared with CC homozygotes, the increase in risk was 3.4-fold in patients with at least one risk factor (smoking, hypercholesterolaemia, diabetes, systemic hypertension) and 4.1 fold in patients with angiographically diagnosed two- or three-vessel disease. We conclude that the GP Ia 807 TT (873 AA) genotype is associated with an increased risk of acute coronary syndrome in the Italian population; conversely, the GP Ia 807 CC (873 GG) genotype seems to represent a protective factor. PMID- 11472361 TI - Decreased fibrinolytic potential in South Asian women with ischaemic cerebrovascular disease. AB - To investigate gender differences in conventional, coagulation and fibrinolytic factors in South Asian ischaemic stroke patients, we compared these variables in 50 South Asian females (SAFP) with 90 South Asian males (SAMP) with ischaemic stroke and in 52 females (SAFC) and 38 males (SAMC) without stroke. Plasminogen activator inhibitor-1 (PAI-1) antigen levels were significantly higher in SAFP compared with SAMP (18.2 vs. 13.3 U/ml, P = 0.04) even after adjustment for known covariates, but there was no difference in PAI-1 antigen levels between males and females in the control group. South Asian females exhibited higher levels of factor VII antigen and FVII:C activity in both stroke patients (114 vs. 99% in males, P = 0.01; 116 versus 104% in males, P = 0.04) and controls (116 vs. 97% in males, P = 0.004; 115 vs. 93% in males, P = 0.01). There were no significant differences in the levels of fibrinogen (3.8 vs. 3.7 g/l), FXIIa (2.2 vs. 2.4 ng/ml), von Willebrand factor (1.8 vs. 1.9 IU/ml) and tissue plasminogen activator (11.4 vs. 12.0 ng/ml) in SAMP and SAFP respectively. These results suggest that South Asian females have increased FVII levels and that females with a history of ischaemic stroke have a decreased fibrinolytic potential in comparison with males. PMID- 11472362 TI - Post-menopausal hormone therapy and concentrations of protein C and antithrombin in elderly women. AB - The effects of post-menopausal hormone therapy (HRT) on blood coagulation in elderly women are not well defined. We studied associations of HRT use with levels of natural anticoagulant proteins in a cross-sectional study of 3393 women > or = 65 years of age participating in the Cardiovascular Health Study. Protein C antigen and antithrombin were measured in all users (n = 230 unopposed oestrogen; 60 oestrogen/progestin) and a comparison group of 196 age- and race matched non-users. Compared with non-users, oestrogen use was associated with higher protein C (4.80 vs. 4.30 microg/ml, P < 0.01). Results were similar for oestrogen/progestin (P > 0.05). In both user groups, antithrombin was lower than in non-users (109% for each vs. 115% in non-users, P < 0.001). Adjustment for factors related to prescription of HRT and to anticoagulant protein levels had little impact on the results. For antithrombin, associations with HRT were larger for thinner Caucasian women and black women. Venous thrombosis from HRT may be mediated partly by alterations in antithrombin, but not protein C concentrations. This study extends previous observations to older women, the group at highest risk of venous thromboembolism. Studies of HRT-induced alterations in anticoagulant function in relation to the occurrence of thrombosis with HRT are required. PMID- 11472363 TI - Protein Z in ischaemic stroke. AB - Many risk factors associated with ischaemic stroke are known, including high levels of fibrinogen or factor VII. Protein Z is a vitamin K-dependent coagulation factor, which was found to promote the assembly of thrombin with phospholipid vesicles that might promote coagulation. Indeed, a low protein Z level may be associated with a varying bleeding tendency. Therefore, we hypothesized that high protein Z levels could induce a hypercoagulable state and performed a case-control study to investigate a potential association between high protein Z plasma levels and ischaemic stroke. We measured protein Z in plasma samples from 157 patients with stroke of unknown aetiology and 192 control subjects. All patients had survived an ischaemic stroke or transient ischaemic attack (TIA) for at least 2 months. We found an increased relative risk of ischaemic stroke with increasing protein Z levels, with an odds ratio of 4.3 [95% confidence interval (CI): 1.7--11] for protein Z plasma levels > or = 160%. Excluding patients with a history of venous thromboembolism from the analysis, the same result was obtained (odds ratio 4.2; 95% CI: 1.6--11.2). Using a logistic regression model, this association also remained significant (P = 0.04) after adjustment for established risk factors. Our data indicated that a high plasma level of protein Z is an independent risk factor for ischaemic stroke. PMID- 11472364 TI - Successful treatment of severe intra-abdominal bleeding associated with disseminated intravascular coagulation using recombinant activated factor VII. AB - Recombinant activated factor VII (rFVIIa) is indicated mainly for the treatment of patients with haemophilia and inhibitors. However, little information is available on the use of rFVIIa in the treatment of the severe bleeding associated with disseminated intravascular coagulation (DIC). We report a pregnant woman with DIC, who developed severe intra-abdominal bleeding after caesarean section. Despite treatment with fresh-frozen plasma, fibrinogen, platelet transfusions and surgery, the abdominal bleeding persisted and intravenous treatment with rFVIIa was initiated. The response to treatment was rapid, with control of the bleeding and resolution of the coagulopathy. No side-effects related to rFVIIa were noted. This case suggests a potential role for rFVIIa in the treatment of severe and refractory bleeding associated with DIC. PMID- 11472365 TI - Evidence that natural killer cells express mini P-glycoproteins but not classic 170 kDa P-glycoprotein. AB - Several lines of evidence including reverse transcription polymerase chain reaction, immunoreactivity and their ability to efflux rhodamine 123 have implied the existence of P-glycoprotein in natural killer (NK) cells. It has been a natural tendency to assume that NK-cell P-glycoprotein is identical to the P glycoprotein of multidrug resistant (MDR) cell lines, however, the present study uncovered major differences. Functionally, NK cells demonstrated a restricted substrate profile, being unable to transport daunorubicin and calcein acetoxymethylester while efficiently transporting other P-glycoprotein substrates. Furthermore, physical differences in NK-cell P-glycoprotein were established by differential reactivity with P-glycoprotein antibodies. NK cells demonstrated strong reactivity with C494 and JSB-1, but did not react appreciably with C219. In addition, NK cells were unable to bind to the antibody MM4.17 unless they had been fixed and permeabilized, yet this antibody normally recognizes an extracellular epitope of P-glycoprotein. These differences culminated in the demonstration using Western analysis that NK cells did not express detectable levels of 170 kDa P-glycoprotein. Instead, NK cells expressed small-molecular-weight 'mini P-glycoprotein' products, of approximately 70 and 80 kDa. Collectively, these data indicate that the predominant P-glycoprotein species of NK cells are novel mini P-glycoproteins and not the classic P glycoprotein of MDR models. PMID- 11472366 TI - (99m)Tc-HMPAO (Ceretec) is stored in and released from the granules of eosinophil granulocytes. AB - (99m)Tc-HMPAO (Ceretec) labelling of leucocytes is used clinically for the detection of inflammatory processes in the body. This study investigated the mechanisms by which (99m)Tc-HMPAO is taken up by eosinophils and neutrophils. Blood cells were labelled with (99m)Tc-HMPAO and the cells separated by means of their densities in Percoll gradients. For other purposes, eosinophils and neutrophils were purified by means of the MACS system and, after labelling these pure cellular preparations, the cells were ultrasonicated and the organelles separated on sucrose density gradients by means of ultracentrifugation. Organelles were characterized by their morphology on electron microscopy. Granulocytes were stimulated to secrete their granule constituents by means of exposure to complement-coated particles. ECP (eosinophil cationic protein) and MPO (myeloperoxidase) were measured using specific immunoassays. The uptake of (99m)Tc-HMPAO was 15--25-fold higher in eosinophils than in other leucocytes. (99m)Tc-HMPAO was predominantly stored in the secretory granules of eosinophils and released from the eosinophil, upon activation, together with ECP. A second storage compartment was a very light density organelle of unknown nature. These results indicated that, among leucocytes, (99m)Tc-HMPAO is preferentially taken up by eosinophils and stored in the secretory granules, which has to be taken into consideration when evaluating images based on this technique. Our findings suggest that (99m)Tc-HMPAO (Ceretec) may be used as a tool to follow eosinophil turnover and activity in disease. PMID- 11472367 TI - Profiling of genes expressed in human monocytes and monocyte-derived dendritic cells using cDNA expression array. AB - Using a human cDNA expression array, we obtained expression profiles of 588 genes in CD14+ monocytes and monocyte-derived dendritic cells (DCs). Overall, 22 genes were upregulated, and nine genes were downregulated in DCs of both samples from two different individuals. Many of the genes that were upregulated in DCs encode proteins that are related to differentiation, cell structure, migration, termination of cell cycle as well as proliferation, e.g. tumour necrosis factor alpha (TNF-alpha), tumour necrosis factor receptor II (TNFRII), thymosin beta-10, epithelial discoidin domain receptor 1, replication factor C, putative transcription factor DB1, alpha catenin, transforming growth factor-beta 1, prohibitin, p53-regulating protein and neu differentiation factor. Among the downregulated genes in DCs were genes that encode proteins of cell cycle regulation: mitotic growth and transcription activator, platelet-derived growth factor receptor-beta subunit, interleukin 2 receptor (IL-2R)-gamma subunit, IL-7R alpha subunit, leucocyte interferon-gamma (IFN-gamma) and granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR). Semi-quantitative reverse transcription-polymerase chain reaction method confirmed the upregulated expression levels in DCs for TNFRII, TNF-alpha, alpha catenin and downregulation of IFN-gamma, GM-CSFR on four different donor samples of DCs and monocytes. Moreover, our data show the presence of a 'switch-on' step for the TNF-alpha and TNFRII gene expression in immature DCs for further differentiation into mature DCs. PMID- 11472368 TI - Autoimmune neutropenia in pregnant women causing neonatal neutropenia. AB - Autoimmune neutropenia (AIN) can occur during pregnancy. However, neonatal neutropenia occurring in an infant born to a mother with AIN has only rarely been documented. Recently, we have experienced two cases of AIN during pregnancy, both of which caused severe yet transient neonatal neutropenia (< 0.3 x 10(9)/l), probably as a result of transplacental maternal anti-neutrophil autoantibodies. The anti-neutrophil antibodies seemed to be against antigens other than NA1/NA2 because the autoantibodies did not bind to neutrophils of specific NA types selectively in the granulocyte indirect immunofluorescence test. Although AIN is a relatively uncommon disease, neonatal neutropenia caused by maternal AIN may not be quite as rare. PMID- 11472369 TI - An in situ study of CD34(+) cells in human fetal bone marrow. AB - The purpose of this study was to characterize the spatial distribution, number and size of CD34(+) cells in fetal bone marrow. Thin sections of normal fetal bone marrow from lumbar vertebrae were stained using CD34 antibody QBend/10. Sections were used under light microscopy with various eyepiece graticules to make measurements of CD34(+) cells in situ. Results showed that at mid- and late gestation, approximately 2% and 0.5% of fetal bone marrow cells were CD34(+) respectively. The mean distance of CD34(+) cells from the nearest trabecular bone surface was 61 +/- 4 and 46 +/- 4 microm, respectively, for mid- and late gestation. The mean distance to the nearest neighbour was 46 +/- 5 and 105 +/- 15 microm, and the mean distance to the nearest blood vessel was 13 +/- 1 and 17 +/- 2 microm respectively. The concentration of CD34(+) cells in the peripheral region was 6.5 times greater than that at the centre of the sections. Overall, the percentage number of CD34(+) cells decreased with gestational age. The cellular and nuclear diameters of CD34(+) cells remained unchanged throughout mid and late gestation at 5.4 +/- 0.1 and 3.8 +/- 0.1 microm respectively. This information will be used to calculate the natural background alpha-radiation dose to haemopoietic stem cells. PMID- 11472370 TI - Variables to predict engraftment of umbilical cord blood into immunodeficient mice: usefulness of the non-obese diabetic--severe combined immunodeficient assay. AB - Umbilical cord blood is an alternative stem cell source for patients without matched family donors. In this study, we examined several parameters that have not been studied in detail -- radiation dose, cell dose, age of mice, and maternal and neonatal characteristics of the cord blood donor -- that affect engraftment of cord blood in non-obese diabetic-severe combined immunodeficient (NOD--scid) mice. Engraftment, measured using flow cytometry analyses of human CD45(+) cells, was highest in 400 cGy-treated mice. Successful engraftment was demonstrated up to 6 months, with a mean engraftment of 31% (range 0--67%) of human cells in recipient bone marrow. Engraftment was skewed to B lymphocytes. The radiation dose of 350 cGy resulted in superior survival of the murine recipients compared with 400 cGy (P = 0.03). The sex of the NOD--scid recipients had a significant effect on survival (female superior to male, P = 0.01), but not on engraftment. There were high levels of variability among different cord units and among animals injected with the same cord unit. This variability may limit the clinical usefulness of the NOD--scid mice as hosts for the quantification of human stem cells. PMID- 11472371 TI - Analysis of chimaerism in thalassaemic children undergoing stem cell transplantation. AB - We have prospectively assessed the relative contribution of host and donor to haemopoiesis following stem cell transplantation (SCT) in children with beta thalassaemia major (n = 35), using karyotype analysis or Southern blot/polymerase chain reaction analysis of variable number tandem repeats on genomic DNA from peripheral blood. Early haemopoiesis was fully donor in origin in 24 out of 35 cases and remained so throughout the post-transplant course in all but one patient, who evolved to stable mixed chimaerism. The remaining 11 cases (31%) initially showed mixed chimaerism: four of these rejected, one eventually eradicated host haemopoiesis to become fully donor haemopoietic, and the remaining six had persistent mixed chimaerism, with 5--38% host haemopoiesis. The risk of graft rejection was high when > 15% host haemopoiesis was present at 3 months post transplant: four out of six such patients rejected their grafts; conversely, zero out of 29 patients with < 15% host haemopoiesis at 3 months rejected (P < 0.0001). There was a higher incidence of significant acute and chronic graft-versus-host disease in patients with full donor chimaerism. These studies confirm that the mixed chimaeric state is common following SCT for thalassaemia, often persists (with up to 4 years follow-up) and is compatible with long-term cure. Analysis of chimaerism in patients undergoing SCT for beta thalassaemia enables monitoring of engraftment in the early post-transplant period, provides insight into the biology of engraftment and may be useful in identifying patients at high risk of rejection. PMID- 11472372 TI - Reduced serum ceruloplasmin levels in hereditary haemochromatosis. AB - As ceruloplasmin (Cp) seems to be involved in iron mobilization, serum Cp levels were measured in 35 patients with hereditary haemochromatosis (HH), 12 with acquired iron overload (AIO) and 36 healthy subjects. Cp was lower in HH patients than in controls (P < 0.001); no difference was found between untreated HH patients and those on a phlebotomy programme (P = 0.07) and between the HH patients carrying the normal and the mutated alleles of the HFE gene (P = 0.8). Cp levels in AIO subjects were significantly higher than in HH patients (P < 0.004) and similar to those of controls (P = 0.2). No differences in albumin, alpha1 acid glycoprotein and copper serum levels were observed in the three groups. PMID- 11472373 TI - Haemolytic onset of Wilson disease in a patient with homozygous truncation of ATP7B at Arg1319. AB - We describe a 19-year-old woman with haemolytic anaemia and thrombocytopenia as the initial manifestation of Wilson disease (WD). There are two reasons for reporting such an improbable case. First, it emphasizes the importance of recognizing atypical clinical presentations of potentially lethal recessive traits for which therapy is available. Second, it shows that, even in a monogenic disorder like WD, the phenotype cannot be extrapolated from the mutated genotype in a simple fashion; this patient had a relatively late-onset form of WD despite homozygosity for a genetic lesion leading to an apparent complete loss of function of the WD copper transporter. PMID- 11472374 TI - The effect of leucocyte depletion on the quality of fresh-frozen plasma. AB - The aim of this study was to evaluate the quality of leucodepleted (LD) fresh frozen plasma (FFP) produced using one of five whole blood filters (Baxter RS2000 & RZ2000, NPBI T2926, Macopharma LST1 and Terumo WBSP) or two plasma filters (Pall LPS1 and Baxter FGR7014). Whole blood or plasma was filtered within 8 h of collection at an ambient temperature. Samples were taken pre- and post filtration for analysis of coagulation factors and complement activation (n = 7--12 for each type of filter). All filtered units (209--286 ml) contained < 5 x 10(6) residual leucocytes and < 30 x 10(9)/l platelets. Statistically significant losses of factors V, VIII, IX, XI and XII and increases in markers of coagulation activation were observed (0--21%), which were dependent on filter type. None of the filters had a significant effect on von Willebrand factor (VWF) multimeric distribution or the activity of VWF and factors II, VII or X. The effect on levels of C3a appeared to be related to the filter surface charge: positively charged filters resulted in C3a generation, whereas negatively charged resulted in C3a removal. None of the observed changes are likely to be clinically significant unless subsequent processing of plasma (such as pathogen inactivation) results in further losses of coagulation factors. PMID- 11472375 TI - Testing Sokal's and the new prognostic score for chronic myeloid leukaemia treated with alpha-interferon: comments. PMID- 11472377 TI - TFR2 Y250X mutation in Italy. PMID- 11472378 TI - Treatment of refractory autoimmune haemolytic anaemia with anti-CD20 (rituximab). PMID- 11472379 TI - Thalidomide and low-dose dexamethasone in myeloma treatment. PMID- 11472380 TI - Use of recombinant factor VIIa for post-operative haemorrhage in a patient with Glanzmann's thrombasthenia and human leucocyte antigen antibodies. PMID- 11472381 TI - Migraine and function of the immune system: a meta-analysis of clinical literature published between 1966 and 1999. AB - Mechanisms underlying migraine precipitation are largely unknown. A role of the immune system in migraine precipitation is a matter of debate because of the association of atopic disorders and migraine. Recently, it was demonstrated that migraineurs benefit from eradication of a Helicobacter pylori infection, which substantiates a possible role for (sub-clinical) infections in precipitation of migraine. Since 1966, about 45 clinical investigations have reported on alterations of immune function in migraine patients, which we present in this review. Changes of serum levels of complement and immunoglobulins, histamine, cytokines and immune cells were found in some of these studies but in most cases not corroborated by others. Migraineurs suffering from comorbid atopic disorders show elevated plasma IgE levels but not patients without a type I hypersensitivity. Histamine plasma levels are chronically elevated in migraineurs, and interictally decreased lymphocyte phagocytotic function and increased plasma tumor necrosis factor alpha (TNFalpha) levels were found, and may be related to increased infection susceptibility. The cause of this increased susceptibility is unclear but most likely is a result of chronic stress, a well known suppressor of the immune system. Stress relief enhances immune activity and triggers a burst of circulating vasoactive compounds that function as mediators of inflammation and potential precipitators of a migraine attack in vulnerable subjects. In conclusion, in the clinical literature of the past decades, there is no clear-cut evidence of an immune dysfunction in migraineurs, but we cannot totally exclude the possibility of an altered immune function in migraineurs. Discrepancies in the literature most likely are caused by the divergent patterns of sample collection relative to the time of the attack. We propose stringent definition of sample collection times for future studies of immune function in migraine patients. PMID- 11472382 TI - Clinical presentation of transformed migraine: possible differences among male and female patients. AB - Chronic daily headache (CDH) represents a group of non-paroxysmal headache disorders that occur on a daily or near-daily basis, for longer than 6 months. Even though it is a common problem, affecting 30-70% of the patients attending specialized headache clinics, it is not a well-defined and classified disorder, resulting in controversies regarding its description and approach. The aim of this study was to evaluate the clinical presentation of CDH due to transformed migraine and possibly compare the differences among male and female patients. Two hundred and seventy-one patients, 217 women and 54 men, ages 16-83 (mean 37.5 years for women and 41.4 for men), fulfilling the proposed criteria for transformed migraine and selected from a group of 300 consecutive CDH patients attending a subspecialty headache centre, were studied retrospectively. The most observed clinical presentation was pressure or tightening, bilateral fronto temporal, moderate non-continuous headache, with a progressive onset. The association with nausea and phonophobia was demonstrated in 60% and 32% of the patients, respectively. The association with photophobia (29.6% male, 44.2% female, P = 0.05) sleep (77.7% male, 49.8% female, P = 0.0002) and emotional (87% male, 64.1% female, P = 0.001) disturbances, as well as the occurrence of intermittent full-blown migraine attacks (81.5% male, 95.4% female, P = 0.001) was significantly different among male and female patients. Overuse of symptomatic medications (SM) was observed in 87% of the male and in 83.8% of the female patients, with a significant difference concerning the use of more than one type of SM (male 68% compared with female 91.7%; P = 0.006). We concluded that TM patients have a clinical presentation compatible with previous descriptions but suggesting, even though limited by the restricted number of male patients, different aspects among male and female patients. PMID- 11472383 TI - Platelet 3H ketanserin binding in migraine. AB - Platelet 3H ketanserin binding was studied in 33 patients of migraine and 30 healthy controls. The binding characteristics: equilibrium dissociation constant (Kd) and maximal number of binding sites (Bmax) determined by Scatchard analysis revealed a significant decrease in Kd and no change in Bmax in migraine cases. No correlation was observed between the Kd and Bmax with the clinical features of migraine. The findings of the present study show that there is a decreased affinity of platelet 5-HT2 receptors in migraine. PMID- 11472384 TI - Cervicogenic headache: evaluation of the original diagnostic criteria. AB - A variety of headaches are frequently associated with the occurrence of neck pain. The purpose of this paper was to describe the adherence to diagnostic criteria of a series of patients enrolled on the basis of two clinical criteria: (1) unilateral headache without side-shift, and (2) pain starting in the neck and spreading to the fronto-ocular area. One hundred and thirty-two patients (36 male and 96 female) entered the study. Sixty-two patients were assigned to Group A (patients fulfilling criteria 1 and 2), 40 to Group B (criterion 2 only) and 12 to Group C (criterion 1, only). Eighteen subjects were excluded because X-rays of the neck were not available. Patients were evaluated regardless of whether or not they fell into one or more of the following diagnostic categories: cervicogenic headache (CEH), migraine without aura (M) and headache associated with disorders of the neck (HN) (IHS definitions). Fulfillment of the diagnostic criteria for CEH was found to be particularly frequent in Group A. A higher frequency of CEH diagnosis was found when two criteria were used (Group A) than in Group B (P = 0.001); in the former group a higher mean number of diagnostic criteria for CEH were also present (P = 0.001). Group A patients more frequently presented pain episodes of varying duration or fluctuating, continuous pain and moderate, non excruciating, non-throbbing pain than Group B patients (P = 0.04 and P = 0.08, respectively). In Group C patients, the frequency of these two criteria was relatively low (17%) especially of the first mentioned variable. The presence of at least five of the seven 'pooled' CEH criteria (present in > or = 50% of the patients) might be deemed a reliable cut-off point, allowing the headache to be diagnosed as 'probable' CEH. If patients fulfilling M or HN criteria in addition to the CEH criteria are added to the 'pure' CEH group a total of 74% of Group A patients may have a CEH picture. The temporal pattern of pain and the quality of pain in Group A showed good sensitivity and specificity (> or = 75) when compared with Group B; therefore, the chances of diagnosing a definite CEH are significantly more frequent in patients presenting with unilateral pain that also begins as a neck pain. Head/neck trauma and radiological abnormalities in the cervical spine were not significantly associated with CEH, M or HN diagnoses. An improvement of the current diagnostic IHS criteria might make it possible to avoid the existing, partial overlap of CEH with HN and M. Extensive use should be made of the GON, and other, blockades in the routine work-up of CEH, both in the differential diagnosis and in the mixed forms (CEH + M, and CEH + HN), in order to improve the efficiency of the current diagnostic system. PMID- 11472385 TI - Diagnosis of migraine: empirical analysis of a large clinical sample of atypical migraine (IHS 1.7) patients and proposed revision of the IHS criteria. AB - The International Headache Society (IHS) diagnostic criteria for headache improved the accuracy of primary headache diagnoses, including migraine. However, many migraineurs receive an 'atypical migraine' diagnosis according to the IHS nosology (IHS 1.7), indicating that they approximate but do not fully meet all IHS criteria. This study characterized and sub-classified patients with atypical migraine. Within a clinical sample of 382 headache sufferers, 83 patients met IHS criteria for 'atypical migraine'. Patients receiving the IHS 1.7 designation did not converge to form a homogeneous group. Rather, distinct and clinically relevant subgroups were empirically derived (e.g. migraine with atypical pain parameters, brief migraine, chronic migraine). The results call for revisions of the IHS diagnostic criteria for migraine that would minimize the number of patients receiving an atypical diagnosis. Revisions would include decreasing the minimum headache duration criteria from 4 h to 2 h, and developing a classification for 'chronic migraine' for migraine greater than 15 days per month. The proposed revision provides a means of diagnosing the daily and near daily headache commonly observed in clinical populations. PMID- 11472386 TI - Patterns of ergotamine and sumatriptan use in the Netherlands from 1991 to 1997. AB - The objective of this study was to assess usage patterns of ergotamine and sumatriptan over a period of 6 years, primarily to evaluate the impact that sumatriptan has had on the prescription of ergotamine. This study used ergotamine and sumatriptan prescription data representing inhabitants of eight cities in the Netherlands and covering the period of 1991-1997. The yearly incidence of new users between 1991 and 1997 was estimated for both drugs as well as for the drug of first choice to be prescribed to patients initiating specific abortive migraine treatment with either ergotamine or sumatriptan. Intra-individual ergotamine and sumatriptan usage patterns, characterized by single (incidental), continuous (rate of retention) or switch use, were examined for five patient cohorts, each for a follow-up period of 1 year. During the year of sumatriptan introduction (1991-1992), the overall incidence of new use for both drugs was highest (5.4 per 1000 inhabitants). Hereafter, a substantial reduction of more than 50% was observed. From 1992 to 1996, the yearly incidence of ergotamine first-time use was significantly higher than that of sumatriptan and up to 1996 ergotamine was more than twice as likely than sumatriptan to be prescribed to patients initiating specific abortive treatment. Hereafter, sumatriptan was as likely as ergotamine to be prescribed as the drug of first choice, which coincided with the full reimbursement of sumatriptan tablets. Overall, neurologists were more likely than general practitioners (GPs), to prescribe sumatriptan as the drug of first choice. Approximately half of the total study population were identified as single-time users. This phenomonen occurred more frequently in the ergotamine cohorts. The sumatriptan cohorts displayed a slight yet significant stronger retention rate compared with the ergotamine cohorts. The overall impact of sumatriptan on ergotamine use in The Netherlands was marginal, predominantly due to GP's adherence to migraine treatment guidelines and reimbursement policies concerning sumatriptan tablets. Overall, incidental use was relatively high and may reflect the reported difficulties in diagnosing migraine, lack of patient-doctor consultation, or that anticipated benefits of the drug were not achieved. Further study is required to clarify these issues. PMID- 11472387 TI - Efficacy and safety of metamizol vs. acetylsalicylic acid in patients with moderate episodic tension-type headache: a randomized, double-blind, placebo- and active-controlled, multicentre study. AB - We assessed the efficacy and safety of oral single doses of 0.5 and 1 g metamizol vs. 1 g acetylsalicylic acid (ASA) in 417 patients with moderate episodic tension type headache included in a randomized, double-blind, placebo- and active controlled, parallel, multicentre trial. Eligibility criteria included 18-65 years of age, history of at least two episodes of tension-type headache per month in the 3 months prior to enrollment, and successful previous pain relief with a non-opioid analgesic. Treatment arms were metamizol 0.5 g (n = 102), metamizol 1 g (n = 108), ASA 1 g (n = 102) and placebo (n = 105). The analgesic efficacy of 0.5 and 1 g metamizol vs. placebo was highly statistically significant (alpha: 0.025; one-sided) for sum of pain intensity differences, maximum pain intensity difference, number of patients with at least 50% pain reduction, time to 50% pain reduction, maximum pain relief and total pain relief. A trend towards an earlier onset of a more profound pain relief of 0.5 and 1 g metamizol over 1 g ASA was noticed. All medications including placebo were almost equally safe and well tolerated. PMID- 11472388 TI - Altered cerebrovascular response pattern in interictal migraine during visual stimulation. AB - A dysbalance of the cerebrovascular response during functional activation of the brain has been postulated as a factor in the pathophysiology of migraine. To determine the dynamic pattern of the cerebrovascular response in migraineurs compared with a control group, changes of the cerebral perfusion during cerebral activation were studied with high temporal resolution by functional transcranial Doppler sonography (fTCD). The cerebral blood flow velocity (CBFV) in the right posterior cerebral artery (PCA) and the left middle cerebral artery (MCA) was measured simultaneously during visual stimulation in 19 interictal migraineurs and in 19 age- and sex-matched control subjects. Data were analysed with a previously validated technique based on automated stimulus-related averaging of the CBFV. The MCA migraineurs exhibited a steady increase of CBFV during the stimulation, while normal subjects showed a habituation of the CBFV response. The lack of habituation in migraineurs was significantly (P < or = 0.05) more pronounced across patients with a high attack frequency (> or = 4 per month) compared with migraineurs with a low attack frequency (< 4 per month). In the PCA, compared with normal subjects, migraineurs showed significantly (P < or = 0.05) stronger CBFV changes at the beginning and after the end of stimulation, with a slower decline to baseline. Data are in accordance with electrophysiological findings in migraineurs. It is assumed that a lack of habituation of the cerebrovascular response in migraineurs might contribute to a disturbance of the metabolic homeostasis of the brain that might induce migraine attacks. PMID- 11472389 TI - Exploding head syndrome and idiopathic stabbing headache relieved by nifedipine. PMID- 11472390 TI - Cluster headache after dental extraction: implications for the pathogenesis of cluster headache? PMID- 11472391 TI - Compulsive yawning as migraine premonitory symptom. PMID- 11472392 TI - Chemokines in myocardial failure -- pathogenic importance and potential therapeutic targets. PMID- 11472393 TI - Enhanced expression and production of monocyte chemoattractant protein-1 in myocarditis. AB - Monocyte chemoattractant protein-1 (MCP-1) is a member of the C-C chemokine family that has been shown to play a major role in the migration of monocytes and T cells to an inflammatory focus. To clarify the role of MCP-1 in the pathogenesis of myocarditis, we have examined the expression of MCP-1 in rat hearts with experimental autoimmune myocarditis (EAM), and have also measured serum levels of MCP-1 in patients with histology-proven acute myocarditis. Lewis rats were immunized with cardiac myosin and were killed 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 42 and 56 days after immunization. Large mononuclear cells in the myocardial interstitium were stained with an anti-MCP-1 antibody. mRNA of MCP-1 increased in the hearts of EAM rats from days 15--27 as shown by quantitative reverse transcription-polymerase chain reaction. Serum MCP-1 levels of the rats with EAM were significantly elevated from days 15--24. In the clinical study, serum levels of MCP-1 in 24 patients with acute myocarditis at the time of admission (165.2 +/- 55.8 pg/ml) were significantly (P = 0.0301) elevated compared with those of 20 healthy volunteers (61.8 +/- 10.7 pg/ml). Serum MCP-1 levels of 8 fatal cases (371.8 +/- 145.2 pg/ml) were significantly (P = 0.0058) higher than those of 16 cases who survived (65.5 +/- 12.8 pg/ml). In conclusions, MCP-1 may play an important role in the pathogenesis of human acute myocarditis as well as in the progression of rat EAM. PMID- 11472394 TI - Increased numbers of circulating donor-specific T helper lymphocytes after human heart valve transplantation. AB - Implantation of cryopreserved human donor heart valves for either congenital or acquired cardiac disease has been performed since the last three decades. Although the clinical outcome is good, long-term valve degeneration resulting in dysfunction has been observed. A specific immunological response of the recipient against the allograft has been proposed as one of the factors involved in this process. Helper T lymphocytes play an important intermediate role in cellular and humoral immune response. Increasing numbers of circulating donor-specific helper T lymphocytes precursors (HTLp) correlate with graft rejection after organ transplantation. To investigate whether cryopreserved human donor heart valves are able to induce a donor-specific T helper response, we monitored the HTLp frequencies (HTLpf) in peripheral blood samples of 13 patients after valve allograft transplantation by use of a limiting dilution assay followed by an interleukin-2 bioassay. Prior to transplantation, HTLpf specific for donor and third-party antigens showed individual baseline levels. After allografting, the antidonor frequencies significantly increased in 11 of the 13 patients (P = 0.02). This was not found for stimulation with third-party spleen cells (P = 0.68), which indicates a donor-specific response. Maximal donor-specific HTLpf were already found at 1--2 months after operation. Valve allograft transplantation induces an increase in the numbers of donor-specific HTLp in peripheral blood of the patients. Analogous to organ transplantation, these HTLp may play a crucial role in events that lead to valve damage. Therefore, monitoring of HTLp in peripheral blood samples might be informative for donor valve degeneration (rejection) and subsequently valve allograft failure. PMID- 11472395 TI - Enhancement of cALL immunogenicity by co-culture with a CD154 expressing 293 cell line. AB - Pre-B cell acute lymphoblastic leukaemia (cALL) commonly occurs in young patients and although successful conventional therapies are available (such as cytotoxic drugs and bone marrow transplantation) for a proportion of patients (approximately 30%) these are ultimately unsuccessful. Recurrence of disease is a result of the failure of the immune system to recognize these abnormal cells and down-regulation of crucial molecules required for cognate CD4(+) T cell recognition has been postulated as a means of immune escape. In this study we show that an embryonic kidney cell line (293 cells) transfected with CD154 (40 L.1) are capable of not only maintaining the viability of primary ALL cells in culture but can also up-regulate the expression of a number of crucial molecules involved in antigen recognition. We show that 40 L.1 cell stimulation of primary ALL cell cultures can not only enhance the allogeneic and autologous MLR response to such cells but will also induce CTL effectors which are capable of lysing wild type autologous ALL cells. It is therefore conceivable that such an approach could be used to generate an active anti-tumour response in patients, following conventional therapy, reducing the incidence of recurrence. PMID- 11472396 TI - Cytolytic complement activity in otitis media with effusion. AB - Otitis media with effusion (OME) is a chronic inflammation persisting in the middle ear cavity of at least 8 weeks duration. Middle ear effusion (MEE; n = 38), samples from children suffering from OME were investigated for their direct cytolytic activity or an ability to enhance complement lysis of unsensitized bystander cells. Thirteen of the 38 MEEs had direct endogenous haemolytic activity and 27 samples had an ability to enhance serum-initiated lysis. Using an enzyme immunoassay, high levels of terminal complement complexes (TCC) were detected in the MEE samples (mean 34.1 microg/ml, range 5--89 microg/ml). This indicated strong local complement activation that had progressed to the terminal stage. As one potential factor promoting complement activation we identified both monomeric and trimeric properdin in MEE by Western blotting. By stabilizing C3 and C5 convertases properdin accelerates the alternative and terminal pathways of complement. On the other hand, the membrane attack complex (MAC) inhibitor CD59, which was found to be extensively shed into the MEE in a functionally active form, may control excessive cytotoxicity of the MEE. In conclusion, intense complement activation, up to the terminal level, maintains ongoing inflammation in the middle ear cavity and can pose a threat to the local epithelium. PMID- 11472397 TI - Identification of antigenic domains on the human sodium-iodide symporter which are recognized by autoantibodies from patients with autoimmune thyroid disease. AB - The sodium-iodide symporter (NIS) is a novel autoantigen in autoimmune thyroid disease. In the present study we have characterized the antigenic domains on the human symporter which are recognized by autoantibodies from patients with either Graves' disease (GD) or autoimmune hypothyroidism (AH). Deletion derivatives of complementary DNA (cDNA) encoding the Na(+)/I(-) symporter were constructed using polymerase chain reaction (PCR) amplification. These deletion constructs were translated in vitro with the concomitant incorporation of [(35)S]methionine into the protein products. The reactivity of seven GD and six AH sera, which were known to contain symporter-binding antibodies, to each of the radiolabelled modified symporters was then determined in immunoprecipitation experiments. Analyses of the results obtained in the radiobinding assays suggest the existence of multiple antibody binding sites on human NIS (hNIS), including regions between amino acids (aa) 1--134, 191--286, 290--411, 411--520 and 520--588. Computer prediction of the potential B cell epitopes on the symporter revealed that, apart from aa 134--191, all the epitope domains identified overlapped, at least in part, with areas predicted to be highly antigenic. Interestingly, the antigenic domains represented by aa 191--286, 290--411 and 411--520 include regions of the polypeptide which form putative extracellular domains in the secondary structure model of the rat symporter. No correlation between the recognition of specific epitopes on the human symporter and the type of autoimmune thyroid disease was demonstrated. PMID- 11472398 TI - Direct B/B-cell interactions in immunoglobulin synthesis. AB - The principal roles of B/B-cell interactions in immune response have not yet been established. We therefore investigated B/B-cell interactions in immunoglobulin synthesis via direct cell-to-cell contact, particularly in the tumour necrosis factor receptor (TNFR)/tumour necrosis factor (TNF) family. We prepared highly purified peripheral blood B cells and stimulated them with Epstein--Barr virus (EBV)-transformed B lymphoblastoid cell lines (LCLs) as activated human B cells. The IgG production by B cells was increased by the addition of fixed LCLs in a dose-dependent manner in the presence of IL-10 plus IL-2. LCLs strongly expressed CD40 and CD70 on their surface, but marginal or no CD154, CD27, OX40 (CD134) and CD134 ligand. The enhancement of immunoglobulin production by LCLs was completely blocked by the initial addition of anti-CD70 blocking MoAb, but not by anti-CD154 or anti-CD134 ligand MoAb. The addition of LCLs also caused a reduction in CD27 expression on B cells, and this effect was completely blocked by anti-CD70 MoAb, indicating a direct B cell--LCL contact via CD27/CD70. LCLs markedly promoted B cell differentiation into plasma cells in the presence of IL-10 plus IL-2. These findings demonstrate that direct interactions between B and B cells via CD27/CD70 induce immunoglobulin production by promoting the generation of plasma cells. PMID- 11472399 TI - Association between natural killer cell activity and infection in immunologically normal elderly people. AB - Congenital patients who lack natural killer (NK) cell activity experience repeated polymicrobial infections. NK cell activity varies significantly among normal people, but it is unknown whether this variation influences their ability to fight infections. This study examined this concern. NK cell activity and other variables, i.e. age, sex, performance status (PS), serum albumin value, lymphocyte and neutrophil counts, various lymphocyte subsets, etc. were determined for 108 immunologically normal elderly subjects who were in nursing homes due to an impaired PS. We analysed for correlations between these variables and the follow-up results of the subjects. Forty-eight subjects developed infection(s) during the first year of follow-up. A low NK cell activity was associated with the development of infection (P = 0.0105, multivariate logistic regression analysis). The relative risk for the development of infection increased in accordance with the decrease in the NK cell activity. Eleven subjects died of infection during the study period. A low NK cell activity was associated with short survival due to infection (P = 0.0056, multivariate Cox's proportional-hazards regression analysis). Our data indicate that low NK cell activity is associated with development of infections and death due to infection in immunologically normal elderly subjects with an impaired PS. PMID- 11472400 TI - Functional T lymphocytes infiltrate implanted polyvinyl alcohol foams during surgical wound closure therapy. AB - Vacuum-assisted closure involving the implantation of polyvinyl alcohol foam is a technique recently developed for the treatment of patients suffering from either wound infection or chronic wounds. This method has been shown to improve and accelerate wound healing. However, little is known about the cell populations that infiltrate the foam, and their potential role in resolving the infection and promoting granulation tissue formation. Our study demonstrates that wound implanted foams are mainly infiltrated with granulocytes, but that mononuclear cells, including macrophages and minor populations of T, B and natural killer lymphocytes, are also present. We show that foam-infiltrating T cells, especially CD4(+) T cells, constitute a phenotypically and functionally heterogeneous population influenced by wound-infecting bacteria. Thus, T lymphocytes could play a role in wound cleansing. In addition, our data indicate that implanted polyvinyl alcohol foams might be suitable microenvironments for manipulating T cell-mediated immune responses in patients. PMID- 11472401 TI - Phenytoin promotes Th2 type immune response in mice. AB - The effects of chronic administration of phenytoin, a common anticonvulsive drug, on immune responses were studied in mice. Anti-keyhole limpet haemocyanin (KLH) IgE antibody response after KLH-immunization was enhanced in phenytoin-treated mice. Proliferative responses of spleen cells induced with KLH, concanavalin A (ConA), lipopolysaccharide and anti-CD3 antibody were reduced in phenytoin treated mice. Accessory function of spleen adherent cells on ConA-induced T cell proliferative response was reduced in phenytoin-treated mice. KLH-induced IL-4 production of spleen cells was enhanced, while IFN-gamma production was reduced in phenytoin-treated mice. In addition, production of IL-1 alpha, but not IL-6 and IL-12 by spleen adherent cells from phenytoin-treated mice was reduced. Natural killer cell activity was reduced in phenytoin-treated mice. These results suggest that phenytoin treatment preferentially induces a Th2 type response. We also observed that plasma ACTH and corticosterone levels were increased in phenytoin-treated mice, and speculated that phenytoin might act directly and indirectly, through HPA axis activation, on the immune system to modulate Th1/Th2 balance. PMID- 11472402 TI - Melatonin provides signal 3 to unprimed CD4(+) T cells but failed to stimulate LPS primed B cells. AB - Growing evidence has supported the conclusion that melatonin, a pineal hormone, modulates the immune function. In our previous study, we evaluated in vivo the potential role of melatonin in the regulation of the antigen specific T and B cells. In the present study, we observe that melatonin down-regulated the expression of the co-stimulatory molecule B7-1 but not B7-2 on macrophages. Further, melatonin encouraged the proliferation of anti-CD3 antibody activated CD4(+) T cells only in the presence of antigen-presenting cells and promoted the production of Th2-like cytokines. Furthermore, it failed to influence the activity of B cells in a T-independent manner. Melatonin suppressed the release of TNF-alpha by LPS or IFN-gamma activated macrophages but failed to inhibit nitric oxide (NO) release. Thus the study shows that melatonin can engineer the growth of unprimed CD4(+) T cells if both the signals are provided by antigen presenting cells. However, it could not regulate the function of B cells. PMID- 11472403 TI - Escherichia coli-induced expression of IL-1 alpha, IL-1 beta, IL-6 and IL-8 in normal human renal tubular epithelial cells. AB - The aim of the present study was to investigate whether the IL-1 family cytokines, in addition to IL-6 and IL-8, could be induced in normal human cortical epithelial cells in response to bacterial stimuli. Human renal tissue was obtained from 9 patients undergoing elective tumour nephrectomy. Renal cortical epithelial cells of tubular origin were prepared from the unaffected tissue. The proximal tubular cells were stimulated for 2, 6 and 24 h with a heat inactivated pyelonephritogenic Escherichia coli strain DS-17. Cultured unstimulated tubular cells served as controls. IL-1 alpha, IL-1 beta, IL-1 receptor antagonist, IL-6, IL-8, IL-10, TNF-alpha, G-CSF and GM-CSF were analysed using immunohistochemistry at the single cell level. The nonstimulated cells were found to express low levels of IL-6 and IL-8 (mean value < 3% of total cells). In contrast, E. coli exposure resulted in significantly increased incidences of IL-6 and IL-8 expressing cells (mean values approximately 18% of total cells) peaking within two hours of stimulation (P < 0.008 and P < 0.02 versus non-stimulated cells, respectively). A gradual decrease was thereafter observed at 6 and 24 h, respectively, although persistently higher compared to controls. A different kinetic response was found for IL-1 alpha, IL-1 beta and IL-1 receptor antagonist expressing cells, which peaked 24 h after E. coli stimulation (mean values 3- 10%) (P < 0.008, P < 0.02, P < 0.02 versus non-stimulated cells, respectively). Low levels of TNF-alpha and GM-CSF were found in 3 of the 9 donated epithelial cells, peaking at 2 h, and IL-10 and G-CSF producing cells in 1 patient each. In conclusion we found that heat-inactivated pyelonephritic E. coli induced a proinflammatory cytokine response in the normal human proximal tubular cells including the IL-1 family, IL-6 and IL-8. PMID- 11472404 TI - The GOR gene product cannot cross-react with hepatitis C virus in humans. AB - GOR (GOR47--1) is an epitope thought to be a host-derived antigen cross-reactive with hepatitis C virus (HCV) since it was isolated from a cDNA library of host animals reactive with sera of HCV-positive patients. An enzyme immunosorbent assay (ELISA) using this epitope as antigen is of sufficient sensitivity and specificity for screening patients with HCV. However, the relationship between GOR47--1 epitope and autoimmune phenomena associated with HCV infection or autoimmune hepatitis is controversial. Here we isolated the human GOR gene and found that the GOR47--1 epitope was not translated in humans due to a single base replacement from chimpanzee. Furthermore, we found some patients who had antibodies against another epitope, which is translated (GOR1--125) in humans, although there was no correlation between the existence of anti-GOR47--1 or anti GOR1--125 Ab and autoimmune phenomena. Serum IgG levels did not influence the titres of these antibodies. Taken together with the results of several other studies, our finding that the GOR47--1 epitope cannot be translated into a protein suggests that there is little relationship between autoimmunity and the GOR gene product in human beings. We also discuss here the possible mechanism of cross-reactivity between HCV and the GOR gene product. PMID- 11472405 TI - High frequency of circulating HBcAg-specific CD8 T cells in hepatitis B infection: a flow cytometric analysis. AB - Viral antigen-specific T cells are important for virus elimination. We studied the hepatitis B virus (HBV)-specific T cell response using flow cytometry. Three phases of HBV infection were studied: Group A, HBeAg (+) chronic hepatitis; Group B, HBeAb (+) HBV carrier after seroconversion; and Group C, HBsAb (+) phase. Peripheral T cells were incubated with recombinant HB core antigen (HBcAg), and intracytoplasmic cytokines were analysed by flow cytometry. HBcAg-specific CD4 and CD8 T cells were identified in all three groups and the number of IFN-gamma positive T cells was greater than TNF-alpha-positive T cells. The frequency of IFN-gamma-positive CD4 and CD8 T cells was highest in Group C, compared with Groups A and B. No significant difference in the HBcAg-specific T cell response was observed between Group A and Group B. The HBcAg-specific CD8 T cell response was diminished by CD4 depletion, addition of antibody against human leucocyte antigen (HLA) class I, class II or CD40L. Cytokine-positive CD8 T cells without HBcAg stimulation were present at a high frequency (7 of 13 cases) in Group B, but were rare in other groups. HBcAg-specific T cells can be detected at high frequency by a sensitive flow cytometric analysis, and these cells are important for controlling HBV replication. PMID- 11472406 TI - Activation of HIV-1-specific immune responses to an HIV-1 vaccine constructed from a replication-defective adenovirus vector using various combinations of immunization protocols. AB - We constructed a recombinant replication defective adenovirus vector containing the env gene (Ad-Bal) derived from macrophage-trophic HIV-1 (HIV-1 Bal). We then immunized mice with this vector using several administration routes and protocols, and examined the immune response. When the Ad-Bal viral vector (over 1 x 10(7) pfu) was injected subcutaneously, both humoral and cell-mediated immunities were induced. However, immune response induced by the Ad-Bal vector alone was weaker than that induced by the recombinant vaccinia viral vector. We then employed the following three immunization protocols: (l) DNA vaccination followed by immunization with the Ad-Bal; (2) vaccination using the Ad-Bal vector followed by DNA vaccination; and (3) DNA vaccination followed by Ad-Bal infection and passive transfer of dendritic cells (DCs) infected with the Ad-Bal. Among the three protocols, the last gave the strongest humoral and cell-mediated immunity. These results suggest that the combination of DNA vaccination, Ad-Bal vector infection and passive transfer of Ad-Bal-infected DCs can induce strong immunity against HIV-1 Bal. PMID- 11472407 TI - Early immune responses accompanying human asymptomatic Ebola infections. AB - In a recent study we identified certain asymptomatic individuals infected by Ebola virus (EBOV) who mounted specific IgG and early and strong inflammatory responses. Here, we further characterized the primary immune response to EBOV during the course of asymptomatic infection in humans. Inflammatory responses occurred in temporal association with anti-inflammatory phase composed by soluble antagonist IL-1RA, circulating TNF receptors, IL-10 and cortisol. At the end of the inflammatory process, mRNA expression of T-cell cytokines (IL-2 and IL-4) and activation markers (CD28, CD40L and CTLA4) was up-regulated, strongly suggesting T-cell activation. This T-cell activation was followed by EBOV-specific IgG responses (mainly IgG3 ang IgG1), and by marked and sustained up-regulation of IFN gamma, FasL and perforin mRNA expression, suggesting activation of cytotoxic cells. The terminal down-regulation of these latter markers coincided with the release of the apoptotic marker 41/7 NMP in blood and with the disappearance of viral RNA from PBMC, suggesting that infected cells are eliminated by cytotoxic mechanisms. Finally, RT-PCR analysis of TCR-V beta repertoire usage showed that TCR-V beta 12 mRNA was never expressed during the infection. Taken together, these findings improve our understanding about immune response during human asymptomatic Ebola infection, and throw new light on protection against Ebola virus. PMID- 11472408 TI - Complete arrest from pro- to pre-B cells in a case of B cell-negative severe combined immunodeficiency (SCID) without recombinase activating gene (RAG) mutations. AB - The B-cell lineage in a patient with B-cell-negative severe combined immunodeficiency (SCID) was analysed by using antisurrogate light chain (SL) MoAbs. Peripheral CD3(+) T cells and CD19(+) B cells were absent in the patient. The common gamma (gamma c) chain was expressed normally on the patient's peripheral NK cells and his peripheral mononuclear cells did not possess any mutations in recombinase activating gene (RAG)-1, 2. Normal levels of expression of Ku70 and Ku80 protein were found by Western blot analysis. The patient did, however, display an increase in fibroblast sensitivity to irradiation. Furthermore, flow cytometric analyses of bone marrow cells showed that surface IgM and cytoplasmic mu positive cells were absent and that CD19(+) B cells were composed of only CD34(+) terminal deoxynucleotidyl transferase (TdT)(+) SL(+) pro B cells. The complete arrest of pro- to pre-B cell development in the SCID patient's bone marrow suggests that some genes involved in V(D)J recombination, excepting the RAG gene, may play a causative role in the immunodeficiency. PMID- 11472409 TI - Assessment of male CVID patients for mutations in the Btk gene: how many have been misdiagnosed? AB - The presentation of hypogammaglobulinaemia in young males without a family history of immunodeficiency can pose a diagnostic problem. In the past, the presence of B-cells has suggested a diagnosis of common variable immunodeficiency (CVID), although genotypic analysis has now clarified that individuals with B cells may have mutations in their Btk gene. In order to address the issue of how many male individuals with a clinical diagnosis of CVID do in fact have mutations in the Btk gene, we analysed a group of 24 male patients. Single-strand conformation polymorphism (SSCP) analysis was used to screen the patient cohort for mutations in the Btk gene. Given the size of the Btk gene, the number of patients in the cohort and the amount of available DNA, multiplex PCR reactions were utilized to span the 19 exons and promoter region of the gene. Where abnormal migration patterns were observed with multiplex PCR reactions, in nine of the 24 patients, the individual Btk gene fragments were re-amplified and analysed again by SSCP. Following this analysis, four patients continued to demonstrate abnormal SSCP migration patterns. However, direct sequencing of the relevant Btk gene fragments for these four CVID patients revealed a mutation in only one patient. The mutation was the previously described polymorphism at position 2031 of Btk gene within exon 18. These results indicate that caution should be taken with the application of SSCP analysis to mutation detection. While it has a role to play in screening large patient cohorts, direct sequencing is a necessary adjunct to such analysis. Finally, the clinical diagnosis of CVID in this cohort successfully excluded males with Btk mutations. PMID- 11472410 TI - Antilactoferrin antibodies in autoimmune liver disease. AB - Antilactoferrin antibodies have been reported in patients with several autoimmune disorders, including primary biliary cirrhosis, autoimmune hepatitis and autoimmune cholangitis. We investigated the prevalence and the clinical significance of such autoreactivity in patients with autoimmune and viral chronic liver disease. Sera from 39 patients with autoimmune hepatitis, 51 with primary biliary cirrhosis, 17 with autoimmune cholangitis, 24 with primary sclerosing cholangitis and 28 with HCV-related chronic hepatitis were studied. Positivity for antilactoferrin antibodies was evaluated by Western immunoblotting with purified human lactoferrin. Antilactoferrin antibodies were detected more often in autoimmune liver disorders (25% autoimmune hepatitis, 25% primary biliary cirrhosis, 35% autoimmune cholangitis, 29% primary sclerosing cholangitis) than in HCV-related chronic hepatitis (3.5%, P < 0.02 versus all). Positivity for antilactoferrin antibodies was not associated with a particular clinical or biochemical profile of the underlying liver disease. No correlation was observed between antilactoferrin reactivity and perinuclear antineutrophil cytoplasmic antibodies. Antilactoferrin antibodies are present significantly more often in autoimmune than in viral liver disorders, but they cannot be considered the serological marker of a specific autoimmune liver disease. PMID- 11472411 TI - Elevated serum L-selectin levels and decreased L-selectin expression on CD8(+) lymphocytes in systemic sclerosis. AB - L-selectin is expressed on most circulating leucocytes and mediates leucocyte rolling on endothelium at sites of inflammation. Following rolling or activation of leucocytes, cell surface L-selectin is released as soluble L-selectin (sL selectin). In the present study, we assessed serum levels of sL-selectin by ELISA and blood leucocyte L-selectin expression by flow cytometry in patients with systemic sclerosis (SSc). Serum levels of sL-selectin in patients with SSc (n = 51) were significantly higher than in normal controls (n = 30) while sL-selectin levels were similar for systemic lupus erythematosus patients (n = 20) and normal controls. Furthermore, SSc patients with elevated sL-selectin levels had inflammatory joint involvement, pitting scar/ulcers, and diffuse pigmentation more frequently than those with normal sL-selectin levels. The frequency of L selectin(+) population among CD8(+) T cells was significantly decreased in SSc patients (n = 30) compared with normal controls (n = 20), while that among CD4(+) T cells, B cells, monocytes, and neutrophils was similar for SSc patients and normal controls. These suggest that elevated sL-selectin levels and decreased frequency of L-selectin+ CD8(+) T cells in SSc patients may be involved in inflammation associated with SSc. PMID- 11472412 TI - Acetylcholine-evoked calcium mobilization and ion channel activation in human labial gland acinar cells from patients with primary Sjogren's syndrome. AB - Recent evidence has indicated that the salivary gland dysfunction associated with Sjogren's syndrome (SjS) is not necessarily due to immune-mediated destruction of acinar tissue. SjS sufferers may possess substantial reserves of acinar tissue but nevertheless be incapable of maintaining salivary flow rates in the normal range. We have investigated the ability of isolated labial gland acinar cells from SjS patients to fluid secrete by measuring agonist-evoked changes in intracellular Ca(2+) ([Ca(2+)](i)) using fura-2 microfluorimetry and activation of K(+) and Cl(-) channels using the patch-clamp whole cell technique. We can confirm that stimulation with a super-maximal dose of acetylcholine (ACh) increased [Ca(2+)]i equally in both control acinar cells and those derived from SjS patients. However, at submaximal concentrations, the dose-response curve for ACh was shifted to the right by approximately one order of magnitude in acinar cells from SjS patients compared to control acinar cells. Patch-clamp measurements consistent with the presence of Ca(2+)-activated K(+) and Cl(-) conductances were obtained from both control acinar cells and those obtained from SjS patients. Dose-dependent activation of the ion channels by acetylcholine was also right-shifted in acinar cells from SjS patients compared to control cells. Our data show that labial gland acinar cells from SjS patients were capable of responding to agonist stimulation by mobilizing [Ca(2+)](i) and activating K(+) and Cl(-) channels consistent with the requirements of fluid secretion. However, the persistent loss of sensitivity to ACh observed in from SjS patients may account for the lack of saliva production observed in these patients in vivo. PMID- 11472413 TI - Inosine monophosphate dehydrogenase (IMPDH) inhibition in vitro suppresses lymphocyte proliferation and the production of immunoglobulins, autoantibodies and cytokines in splenocytes from MRLlpr/lpr mice. AB - The aim of this study was to evaluate the effects of the immunomodulating drug mycophenolic acid (MPA) on splenocytes in an animal model of systemic lupus erythematosus (SLE), using MRLlpr/lpr mice. MPA reversibly inhibits inosine 5' monophosphate dehydrogenase, an enzyme involved in the de novo guanosine synthesis. Splenocytes were treated with MPA (at 1 or 10 microM), and stimulated with either lipopolysaccharide (LPS; 10 microg/ml) or concanavalin A (ConA; 1.25 microg/ml). In blocking experiments, guanosine (100 microM) was added to the cultures to inhibit the effects of MPA. Lymphocyte proliferation, enumeration of immunoglobulin producing cells (using ELISPOT) and quantification of anti-double stranded (ds) DNA antibodies, IFN-gamma and IL-10 (by ELISA) in supernatants were performed. In addition, cell viability was evaluated using propidium iodide and flow cytometry. We found that MPA-treated splenocytes had dramatically decreased mitogen-induced proliferation and number of immunoglobulin producing cells, down regulated production of IFN-gamma, IL-10 and IgM anti-dsDNA antibodies. The viability of MPA-treated cells was also decreased. All of the effect modulated by MPA could be neutralized by the addition of guanosine. We conclude that MPA has potent immunomodulating effects on both B and T lymphocytes, modulating not only proliferation, but also the production of cytokines, immunoglobulins and autoantibodies. PMID- 11472414 TI - Functional heterogeneity of anti-endothelial cell antibodies. AB - While it has been claimed that some anti-endothelial cell antibodies (AECA) activate EC, there is also evidence that others trigger apoptosis. To address the issue of whether activation is a prerequisite for AECA-mediated apoptosis of EC, 23 AECA-positive sera were evaluated for their ability to induce activation and/or apoptosis. Activation was defined as an over-expression of E-selectin and intercellular adhesion molecule 1. Optical microscopy, annexin V binding, hypoploid cell enumeration, and determination of poly (ADP-ribose) polymerase cleavage-related products were used to assess apoptosis. Four functional profiles were defined: 10 sera promoted activation and apoptosis (act+/apo+), one was act+/apo-, six act-/apo+, and the remaining six act-/apo-. The reduced membrane expression of thrombomodulin was associated with apoptosis, rather than activation. Caspase-3 was implicated in the two models of apoptosis, the ratios of several survival proteins to Bax decreased, regardless of the ability of apo+ AECA to activate the cells, while radical oxygen species did not appear to be involved. Furthermore, it occurred that macrophages engulfed EC treated with apoptosis-promoting AECA, but not those incubated with AECA that did not induce apoptosis. Hence, AECA represent an extremely heterogeneous family of autoantibodies, not only because of the variety of their target antigens, but also the subsequent diversity of their effects. PMID- 11472415 TI - Detection of 'antiphospholipid' antibodies: a single chromogenic assay of thrombin generation sensitively detects lupus anticoagulants, anticardiolipin antibodies, plus antibodies binding beta(2)-glycoprotein I and prothrombin. AB - The diagnosis of the antiphospholipid syndrome (APS) requires both a typical clinical event plus a persistently positive test in an assay for either anticardiolipin (aCL) antibodies or a lupus anticoagulant (LA). Enzyme linked immunosorbent assays (ELISA) specific for autoantibodies against beta(2) glycoprotein I (beta(2)GPI) or prothrombin are also used, but none of the tests are adequately sensitive or specific. A chromogenic assay was developed that measures the effect of test antibody or plasma samples on in vitro thrombin formation. It is able to detect both LA and beta(2)GPI-dependent aCL antibodies and may have greater specificity for APS than currently available tests. Using this method various monoclonal antibodies (MoAbs) were examined, from mice immunized with beta(2)GPI, mice with a spontaneous animal model of APS, and from three humans with APS. Plasma and affinity purified antibodies from patients with APS and control groups were also examined. Thrombin inhibition was more sensitive to perturbation by MoAbs than a combination of tests for LA (P < 0.05) and at lower antibody concentrations (12.5 microg/ml versus 100 microg/ml). There was a significant correlation between inhibition of thrombin generation and the level of MoAb reactivity to beta(2)GPI (r = 0.90; P < 0.001) but not to CL (r = 0.06; P = 0.76). Plasma and affinity purified antibodies from patients with APS also inhibited thrombin generation, and significantly more so than patients with aPL from causes other than APS. APS patient samples showed thrombin inhibition in the presence of anti-beta(2)GPI or antiprothrombin antibodies. All MoAbs binding beta(2)GPI showed inhibition of thrombin generation, while MoAbs binding domain I of beta(2)GPI had more LA effect. PMID- 11472416 TI - Molecular mapping of epitopes on melanocyte-specific protein Pmel17 which are recognized by autoantibodies in patients with vitiligo. AB - Previously, we reported the identification of Pmel17 autoantibodies in some patients with vitiligo. Here, we have determined the B cell epitopes on Pmel17 which are recognized by these autoantibodies. Deletion derivatives of Pmel17 cDNA were constructed using either subcloning of specific cDNA fragments or polymerase chain reaction amplification. Full-length Pmel17 cDNA and its truncated derivatives were then translated in vitro to produce [(35)S]-labelled proteins. The radiolabelled ligands were used subsequently in radiobinding assays to investigate the reactivity of sera from vitiligo patients. Two epitope regions were identified: one located at the C-terminal end of Pmel17 between amino acids 634--644 and one in a central region of the protein between amino acids 326--341. Computer analysis of the potential B cell epitopes on Pmel17 revealed that the epitope domain encompassing amino acids 326--341 was located in an area of the protein which was predicted to be highly antigenic. In contrast, the epitope identified at the C-terminal of Pmel17 (amino acids 634--644) was located in a region of the protein predicted to have low antigenicity. The amino acid sequences of the identified Pmel17 epitopes were compared to the amino acid sequences of the related melanogenic enzymes tyrosinase, tyrosinase-related protein-1 and tyrosinase-related protein-2. However, no sequence homology was found between either of the Pmel17 epitopes and the aforementioned proteins. This finding is consistent with our previous study in which we were unable to show the presence of Pmel17 antibodies which were cross-reactive with either tyrosinase, tyrosinase-related protein-1 or tyrosinase-related protein-2. It also suggests that the IgG response to Pmel17 is distinct from the antibody response to the other melanocyte-specific antigens. PMID- 11472417 TI - Skin T cell proliferative response to M protein and other cell wall and membrane proteins of group A streptococci in chronic plaque psoriasis. AB - To determine and compare the T cell response to M protein and other group A streptococcal (GAS) antigens, T cell lines (TCL) were cultured from the lesional skin of 33 psoriatic patients and 17 disease controls. GAS-reactive skin TCL were tested in proliferation assays with recombinant M6 protein, and extracts of cell wall and membrane from type M6 GAS and its corresponding M gene deletion mutant. Initially, GAS-reactive skin TCL were obtained from 16 of 25 (64%) psoriasis, and from seven of 17 (41%) control patients. Eleven psoriatic and four control GAS reactive TCL proliferated to M6 cell wall extract, whereas all the TCL from both groups responded to the extract of M6 membrane proteins. This difference in response to the two extracts was significant for both groups of patients (psoriasis, P = 0.0335, controls, P = 0.0156). GAS-reactive TCL from a further eight psoriasis patients showed no difference in response to cell wall extract from M6 GAS (containing the M protein minus its C-terminus) compared to that of its corresponding M gene deletion mutant. Furthermore, GAS-reactive TCL did not proliferate to recombinant M6 protein. However, a small, but significant reduction in proliferation by the eight psoriatic GAS-reactive TCL to the M negative (lacking the M protein C-terminus) compared to M6-positive membrane extract was observed (P = 0.01). These findings suggest that GAS-reactive T cells in skin lesions of chronic plaque psoriasis proliferate to streptococcal membrane and, to a lesser extent, cell wall proteins. However, psoriatic skin T cells do not recognize cell wall M protein. PMID- 11472418 TI - Benign autoimmunity to combat malignancy. PMID- 11472419 TI - De-novo humoral immune responses to cancer-associated autoantigens during transition from chronic liver disease to hepatocellular carcinoma. AB - A feature of hepatocellular carcinoma (HCC) is that antecedent liver cirrhosis and chronic hepatitis are common precursor conditions and during transition to malignancy some patients develop autoantibodies which were not present during the preceding chronic liver disease phase. Serum samples from such patients can be used to immunoscreen cDNA expression libraries to identify genes encoding the new autoantigens. We demonstrate here the de novo appearance of antibodies to p62, a cytoplasmic protein which has been shown to bind to a developmentally regulated fetal species of insulin-like growth factor II (IGF-II) mRNA. Another antibody appearing during the transition period was against CENP-F, a cell cycle-related nuclear protein with maximum expression in the G2 and M phases of the cell cycle and previously shown to have a high association with malignancy. In three additional patients in whom serial serum samples were examined, new appearance of anti-p62 was detected in two patients and anti-CENP-F in one patient. This study demonstrates that transition to malignancy can be associated with autoantibody responses to certain cellular proteins which might have some role in tumorigenesis. PMID- 11472420 TI - An abnormal adherence of monocytes to fibronectin in thyroid autoimmunity has consequences for cell polarization and the development of veiled cells. AB - Blood monocytes of patients with thyroid autoimmune disease (TAID) display defects in rearranging their cortical actomyosin cytoskeleton ('polarize') in response to chemoattractants. Such rearrangements also take place after the adherence of monocytes to the extracellular matrix (ECM). It is therefore not surprising that monocytes are primed after fibronectin (FN) adherence, displaying an enhanced polarization toward chemoattractants. We investigated the integrin expression and chemoattractant-induced polarization of monocytes of TAID patients before and after FN adherence. Since cytoskeletal rearrangements are also required during the transition of monocytes into veiled antigen-presenting cells (VCs), we investigated such transition of FN-adherent monocytes of TAID patients. Adherent and nonadherent monocyte populations from TAID patients and healthy controls were subjected to a polarization test with the chemoattractant fMLP (or MCP-1), FACS analyses (FITC-labelled FN, CD29, CD49e, d, b and a) and tested for their capability to develop into veiled APC. Monocytes of healthy individuals showed an improved chemoattractant-induced cell polarization after FN adherence, not reflected by TAID monocytes, in which chemoattractant-induced polarization worsened. Monocytes of healthy individuals up-regulated CD49e and d integrins and their capability to bind FITC-labelled FN after adherence to a FN-coated plate, as well as enhancing their capability to generate T cell-stimulatory VCs. Monocytes of TAID patients did not. These data indicate that integrin- (and chemokine-) mediated functions are hampered in monocytes of TAID patients. Because integrin action is pivotal to processes such as monocyte adherence to endothelial cells, uropod formation, migration into tissues and differentiation into APC and macrophages, these defects might underly immune dysbalances important in thyroid autoimmune development. PMID- 11472421 TI - Novel immunosuppressive effect of FK506 by augmentation of T cell apoptosis. AB - We have recently reported the accumulation of oligoclonal activated T cells in the spontaneously developed autoimmune pancreatitis in aly/aly mouse. In this study, we examined the effects of FK506 in this mouse model in preventing autoimmune pancreatitis and investigated its action on calcium signalling apoptosis of alymphoplasia (aly) lymphocytes in vitro. Mice were treated with FK506 from 8 to 25 weeks of age. At the age of 15 weeks, minimal mononuclear cell infiltration was observed in the pancreas in both the FK506 treated group and the control group. Furthermore, a marked cell infiltration associated with destruction of acini and partial fatty changes were observed in 25-week-old control mice. In contrast, FK506 treated mice showed almost no tissue destruction or mononuclear cell infiltration at the age of 25 weeks. Furthermore, at 15 weeks of age, most mononuclear cells in FK506-treated mice were TUNEL positive, whereas only a few were positive in control mice. This augmentation of T cell apoptosis by FK506 was confirmed using naive splenocytes activated by PMA and ionomycin in vitro. Finally, a suppressive effect of FK506 on Bcl-2 production but not on Bax production was confirmed by Western blotting. This unique effect of FK506 on the augmentation of T cell apoptosis is probably one of the mechanisms explaining its beneficial effect on aly autoimmune pancreatitis. PMID- 11472422 TI - Co-incubation of pig islet cells with spleen cells from non-obese diabetic mice causes decreased insulin release by non-T-cell- and T-cell-mediated mechanisms. AB - In vitro studies were conducted in the non-obese diabetic (NOD) mouse, prone to Type 1 autoimmune diabetes, to investigate the mechanisms involved in cell mediated rejection of pig islet xenografts. Our previous work concerning the mechanisms of proliferation of xenogeneic lymphocytes to pig islet cells (PIC) was not indicative of PIC impairment. Consequently, a test was developed based on perifusion analysis of the alteration of basal and stimulated insulin release from adult PIC incubated with mouse splenocytes or subsets. Compared with PIC incubation alone or with syngeneic pig splenocytes, co-incubation with mouse whole spleen cells resulted in a decrease of basal and stimulated insulin release (P < 0.001). Two components of this alteration were detected separately: PIC impairment was decreased (P < 0.01) after removal of plastic-adherent cells from spleen cells, but maintained (P < 0.01) when plastic-adherent cells alone were co incubated with PIC. The increase of murine interleukin-1 beta when mouse plastic adherent spleen cells were cultured with PIC (P < 0.04) was indicative of macrophage activation. Soluble factors produced during co-incubation of mouse splenocytes or plastic-adherent cells with PIC were involved in the impairment process, since supernatant fluids collected during previous PIC-mouse cell co incubations directly altered (P < 0.01) insulin release from PIC. Moreover, impairment of PIC by mouse spleen cells was abolished (P < 0.01) by gadolinium chloride (which inhibits macrophages), but not by cyclosporin A. Another mechanism was apparent, since co-incubation of PIC with purified mouse T cells or CD4+ T cells, re-mixed with antigen-presenting cells, led to a decrease (P < 0.01) of insulin release. This model, based on the alteration of dynamic basal and stimulated insulin release, is indicative of in vitro cell-mediated alteration of PIC in the NOD mouse. The effect of whole spleen cells was rapid, and a crucial role was played by plastic-adherent cells. Two mechanisms were responsible for the behaviour of these cells: an early direct effect (at least in part via soluble products); and the indirect presentation of PIC xenoantigens (leading to impairment by CD4+ T lymphocytes). PMID- 11472423 TI - Characterization of conditions for the primary culture of human small intestinal epithelial cells. AB - Intestinal epithelial cells (IECs) are important for many aspects of gut physiology and pathology. Different approaches have been tried for the primary culture of human IECs, with varying degrees of success, as apoptosis easily occurs. Our aim was to develop a method for primary culture of human IECs from biopsy material. IECs and Lamina propria (LP) cells were liberated from duodenal biopsies obtained from subjects undergoing routine endoscopy for clinical investigations, whose small bowel was macroscopically normal. IECs were cultured on collagen membranes in a 12-well tissue culture cluster, with LP cells and allogeneic Epstein-Barr Virus (EBV)-transformed B lymphocytes (allo-B cells) underneath, in the well. Cultured IECs were characterized by light and confocal microscopy. Cytokine levels in culture supernatants were measured by ELISA. Cells showed the columnar morphology of IECs, even after several days in culture. Best results were obtained from IECs cultured above both LP and allo-B cells. IECs did not form monolayers as do transformed epithelial cell lines, but they did preserve their original cell-cell contacts. Analysis of culture supernatants showed that IL-10 was produced by IECs initially, but IL-1ra was produced by LP cells in the underlying wells with increasing time in culture. Very little IL-1 beta was produced from any cultures. These results show that IECs can be isolated and maintained in primary culture for a short while, which could open new possibilities for research using patient material instead of cell lines. PMID- 11472424 TI - Oral administration of unmodified colonic but not small intestinal antigens protects rats from hapten-induced colitis. AB - Colonic administration of a hapten, 2,4,6-trinitrobenzene sulphonic acid (TNBS) has been shown to induce colitis in rats. We are using this model to investigate the role of colonic antigens in the immunopathology. In this study, we show that colitis can be suppressed by oral administration of haptenized colonic antigens prior to the TNBS enema. Moreover, our data suggest that haptenization of the colonic antigens is not essential because oral feeding of non haptenized colonic antigens too protects rats from TNBS-induced colitis. Thus, unmodified colonic antigens may be involved in the induction of oral tolerance, and possibly in the pathogenesis in this model of colitis. Further, we show that the protective immunity or oral tolerance induced by non haptenized colonic antigens can be passively transferred to naive rats by mesenteric T lymphocytes. Interestingly, oral feeding of small intestinal antigens, haptenized and non haptenized, does not protect rats from colitis, suggesting a specific role for colonic antigens. These data underscore the usefulness of this rat model in the identification of pathogenic antigens in colitis and in the development of therapeutic strategies based on oral tolerance. PMID- 11472425 TI - Subtractive screening reveals up-regulation of NADPH oxidase expression in Crohn's disease intestinal macrophages. AB - Macrophages play a central role during the pathogenesis of inflammation. In normal intestinal mucosa surface expression of typical macrophage markers such as CD14, CD16, CD11b or T-cell co-stimulatory molecules such as CD80 or CD86 is low indicating anergy and low pro-inflammatory activity of these cells. During inflammatory bowel disease (IBD) the mucosa is invaded by a population of macrophages displaying these markers, secreting higher cytokine levels and representing an activated cell population. CD33(+) cells (macrophages) were isolated from normal and Crohn's disease mucosa and mRNA was isolated by polyT magnetic beads. A subtractive screening was performed subtracting mRNA from normal macrophages from those of Crohn's disease macrophages. Oxidative burst activity was determined by flow cytometry. Seventy clones were obtained by the subtractive mRNA screening. Sequencing showed > 99% homology to mRNA of monocyte chemoattractant protein-1 (MCP-1) for three clones. Five clones obtained by subtraction revealed > 99% homology to mRNA of cytochrome b (subunit gp91). Differential expression of the cytochrome b subunit gp91 and the cytosolic NADPH oxidase subunit p67 was confirmed by RT-PCR and 'virtual' Northern blots. The fluorescence ratio of stimulated versus unstimulated cells was 0.9 +/- 0.16 in control macrophages indicating a lack of oxidative burst activity. In Crohn's disease this ratio was significantly increased to 1.80 +/- 0.8 (P = 0.004) confirming the molecular data. In conclusion NADPH oxidase mRNA is down-regulated or absent in macrophages from normal mucosa correlating with a lack of oxidative burst activity. In IBD macrophage-oxidative burst activity is increased and NADPH oxidase mRNA induced. Inhibition of NADPH oxidase could be a new therapeutical target in IBD and reduce mucosal tissue damage in active IBD. PMID- 11472426 TI - Cooperation of interleukin-17 and interferon-gamma on chemokine secretion in human fetal intestinal epithelial cells. AB - Interleukin (IL)-17 is a newly identified T cell-derived cytokine that can regulate the functions of a variety of cell types. In this study, we investigated the effects of IL-17 and interferon (IFN)-gamma on chemokine secretion in human fetal intestinal epithelial cells. IL-8 and monocyte chemoattractant protein (MCP)-1 secretion by the human fetal intestinal epithelial cell line, intestine 407, was evaluated by ELISA and Northern blot. The expression of IL-17 receptor (R) was analysed by a binding assay using [(125)I]-labelled IL-17. The activation of nuclear factor-kappa B (NF-kappa B), NF-IL6 and AP-1 was assessed by an electrophoretic gel mobility shift assay (EMSA). IL-17 induced a dose-dependent increase in IL-8 and MCP-1 secretion. The inducing effects of IL-17 on IL-8 and MCP-1 mRNA abundance reached a maximum as early as 3 h, and then gradually decreased. IL-17 and IFN-gamma synergistically increased IL-8 and MCP-1 secretion and mRNA abundance. IFN-gamma induced a weak increase in IL-17 R mRNA abundance, and incubation with IFN-gamma for 24 h enhanced [(125)I]-labelled IL-17-binding by 2.4-fold. IL-17 rapidly induced the phosphorylation and degradation of I kappa B alpha molecules, and the combination of IL-17 and IFN-gamma induced a marked increase in NF-kappa B DNA-binding activity as early as 1.5 h after the stimulation. Furthermore, this combination induced an increase in NF-IL-6 and AP 1 DNA-binding activity. In conclusion, it becomes clear that IL-17 is an inducer of IL-8 and MCP-1 secretion by human fetal intestinal epithelial cells. The combination of IL-17 with IFN-gamma synergistically enhanced chemokine secretion. These effects of IL-17 and IFN-gamma might play an important role in the inflammatory responses in the intestinal mucosa. PMID- 11472427 TI - Inhibition of p38 MAP kinase during cellular activation results in IFN-gamma dependent augmentation of IL-12 production by human monocytes/macrophages. AB - Interleukin-12 (IL-12) is a key immunomodulatory cytokine produced by antigen presenting cells that promotes cellular immunity and enables the generation of protective immunity against intracellular pathogens and tumours. Therefore, modulation of IL-12 activity is a primary immunotherapeutic goal. However, little is known about its regulation. Signalling via p38 MAPK has been implicated in the control of inflammatory responses and is therefore a potential therapeutic target. We have used the highly selective p38 MAPK inhibitor (SB203580) to examine the effect of this pathway on the production of IL-12. Surprisingly, we found that SB203580 strongly up-regulated LPS induced IL-12p40 at the protein (intracellular and secreted) and mRNA levels in PBMC cultures. The effect on IL 12 was apparent using both T cell-independent and T cell-dependent stimuli but not in unstimulated cultures, indicating that activation signals are required. Furthermore, the production of IFN-gamma by T cells is crucial as production was not increased in LPS-stimulated, purified adherent monocytes/macrophages without the addition of exogenous IFN-gamma. These results provide evidence that p38 MAPK has an unexpected suppressive effect on IL-12p40 gene transcription, and suggests interplay between p38 MAPK- and IFN-gamma -mediated signals in the regulation of IL-12 production by monocytes/macrophages. Furthermore, the importance of IL-12 as a key immunoregulatory cytokine suggests that the clinical application of pyrinidyl imidazole inhibitors, such as SB203580, may need to be reassessed. PMID- 11472428 TI - Somatostatin receptor (SSTR) expression and function in normal and leukaemic T cells. Evidence for selective effects on adhesion to extracellular matrix components via SSTR2 and/or 3. AB - We have examined normal T-cells and T-cell lines with respect to expression of various somatostatin receptor subtypes (SSTR1--5) using RT-PCR and PCR. To evaluate the function of these receptors we have further studied the effects of subtype specific signalling on T-cell adhesion using somatostatin analogs specific for various receptors as probes. Human T-lymphocytes showed SSTR expression related to activation and stage of differentiation. Normal T-cells (peripheral blood, T-cell clone) and T-leukaemia cell lines expressed SSTR2, SSTR3 and SSTR4. Normal T-cells expressed SSTR1 and SSTR5 while T-leukaemia lines did not. SSTR5 was selectively expressed in activated normal T-cells. T lymphocytes produced no somatostatin themselves. Somatostatin and somatostatin analogs specific for SSTR2 and/or SSTR3 enhanced adhesion of T-cells to fibronectin (FN), and to a certain extent, also to collagen type IV (CIV) and laminin (LAM). T-lymphocytes express multiple SSTR and somatostatin may therefore regulate lymphocyte functions via distinct receptor subtypes as shown here for adhesion to extracellular matrix components (ECM) via SSTR2 and SSTR3. SSTR expression also distinguishes normal and leukaemic T-cells. Our findings suggest that SSTR subtypes may be useful targets for therapy during inflammatory diseases and malignancies affecting lymphocytes. PMID- 11472429 TI - Inter-individual differences in cytokine release in patients undergoing cardiac surgery with cardiopulmonary bypass. AB - Cardiac surgery with cardiopulmonary bypass (CPB) leads to a systemic inflammatory response with secretion of cytokines (e.g. IL-6, TNF-alpha, IL-1 beta and sIL-2R). The objective of the following study was to investigate in vitro and in vivo cytokine responses and white blood cell counts (WBC) of patients with high versus low cytokine secretion after a coronary artery bypass grafting (CABG) procedure. Twenty male patients undergoing elective CABG surgery with CPB under general anaesthesia were enrolled in the study. On the day of surgery (postoperatively), serum levels of TNF-alpha and IL-1 beta were significantly higher in patients of the high IL-6 level group compared to the respective values in the patient group with low IL-6 levels. The inter-individual differences in IL-6 release in patients undergoing CABG surgery with CPB were accompanied by differences in the release of other cytokines, such as TNF-alpha, IL-1 beta and sIL-2R. To understand whether genetic background plays a role in influencing cytokine plasma levels under surgical stress, we examined the distribution of polymorphic elements within the promoter regions of the TNF-alpha and IL-6 genes, and determined their genotype regarding the BAT2 gene and TNF beta intron polymorphisms. Our preliminary data suggests that regulatory polymorphisms in or near the TNF locus, more precisely the allele set 140/150 of the BAT2 microsatellite marker combined with the G allele at -308 of the TNF alpha gene, could be one of the genetic constructions providing for a less sensitive response to various stimuli. Our results suggest: (1) close relationships between cytokine release in the postoperative period, and (2) inter individually varying patterns of cytokine release in patients undergoing CABG surgery with CPB. PMID- 11472430 TI - Plasma cytokine response to surgical stress in schizophrenic patients. AB - Schizophrenic patients are reported to have immunological dysfunction, however, the immune response to surgery in schizophrenic patients remains unclear. We measured plasma interleukin-6 (IL-6), interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-alpha) before, during and after colectomy, hemicolectomy and sigmoidectomy in 25 chronic schizophrenic patients (Group S) and 25 control patients (Group C) using ELISA assays. We could find no significant difference in the baseline plasma concentrations of IL-6, IL-8 and TNF-alpha between Group S and Group C. Plasma IL-6 concentrations (32.1 (30.3) and 15.8 (9.6) pg/ml) in Group S at the end of the operation and 24 h after surgery were significantly lower than 76.9 (37.1) and 35.1 (21.5) pg/ml of Group C. Plasma IL-8 concentration (6.1 (2.8)) in Group S at the end of the operation was significantly lower than 8.7 (4.2) pg/ml of Group C. There were no significant changes in plasma TNF-alpha concentration throughout the study period in either group. Plasma cortisol concentrations of schizophrenic patients during surgery were significantly lower than those of control patients. The plasma IL-6 concentrations correlated with plasma cortisol concentrations in either group. We conclude that proinflammatory cytokine response to abdominal surgery is inhibited in schizophrenic patients. PMID- 11472431 TI - Selection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1. AB - We have prepared human recombinant antibody molecules against the glycoprotein antigen of the rabies virus (GPRV) based on the single chain variable fragment (scFv) format. Anti-GPRV scFvs were selected from a human synthetic scFv phage display library with a repertoire of approximately 109 specificities. After three rounds of selection against the PV11 strain of the virus, 40% of the clones tested recognized the rabies antigen. Of the 20 positive clones that were sequenced, five distinct sequences were identified. These distinct scFvs were cloned into a mammalian expression vector carrying the human IgG1 Fc region. The specificity of the resulting scFv-Fc molecules for GPRV was established by ELISA, dot blot and western blot analyses and membrane immunofluorescence. Two of the scFv-Fc fusion proteins neutralized the PV11 strain in a standard in vivo neutralization assay where the virus was incubated with the scFv-Fc molecules before intracranial inoculation in mice. These anti-GPRV scFv-Fc molecules have the potential to be used as an alternative to the presently available HRIG, for use in post-exposure preventive treatment. PMID- 11472432 TI - Modulation of HIV transcription by CD8(+) cells is mediated via multiple elements of the long terminal repeat. AB - HIV replication and LTR-mediated gene expression can be modulated by CD8(+) cells in a cell type-dependent manner. We have previously shown that supernatant fluids of activated CD8(+) cells of HIV-infected individuals suppress long terminal repeat (LTR)-mediated transcription of HIV in T cells while enhancing transcription in monocytic cells. Here, we have examined the effect of culture of T cells and monocytic cells with CD8(+) supernatant fluids, and subsequent binding of transcription factors to the HIV-1 LTR. In transfections using constructs in which NF kappa B or NFAT-1 sites were mutated, the LTR retained the ability to respond positively to culture with CD8 supernatant fluid in monocytic cells. Nuclear extracts prepared from both Jurkat T cells and U38 monocytic cells cultured with CD8(+) cell supernatant fluid demonstrated increased binding to the HIV-1 LTR at an AP-1 site which overlapped the chicken ovalbumin upstream promoter (COUP) site. In monocytic cells, increased binding activity was observed at the NF kappa B sites of the LTR. In contrast, an inhibition in binding at the NF kappa B sites was observed in Jurkat cells. Examination of two NFAT-1 sites revealed enhanced binding at - 260 to - 275 bp in U38 cells which was reduced by cellular activation. PMA and ionomycin-induced binding at a second NFAT-1 site (- 205 to - 216 bp) was abrogated by CD8(+) cell supernatant fluid in T cells. These results, taken together, suggest that factors present in CD8(+) supernatant fluids may act through several sites of the LTR to modulate transcription in a cell type-dependent manner. PMID- 11472433 TI - Elongation factor 1 beta/delta of Echinococcus granulosus and allergic manifestations in human cystic echinococcosis. AB - Allergic reactions, such as urticaria, itching and anaphylactic shock, often complicate the course of cystic echinococcosis (CE). To investigate the role of the IgE-immunoreactive recombinant Echinococcus granulosus elongation factor-1 beta/delta (EgEF-1 beta/delta) in the allergic disorders during CE we determined humoral and cell-mediated responses to this antigen in patients with CE grouped according to the clinical presence or absence of allergic reactions. Immunoblotting analysis showed that serum IgE-binding reactivity to EgEF-1 beta/delta differed significantly in patients with and without allergic reactions (38 of 42, 90% vs. 31 of 56, 56%; P < 10(-4)). EgEF-1 beta/delta induced a proliferative response in 14 of 19 (74%) patients' peripheral blood mononuclear cells (PBMC) irrespective of the allergic manifestations and skewed Th1/Th2 cytokine activation towards a preferentially Th2 polarization. Epitope mapping identified an immunodominant epitope of 18 residues with 78% identity and 89% similarity with an IgE-immunoreactive Strongyloides stercoralis antigen. Overall these findings suggest that EgEF-1 beta/delta is an allergenic molecule that may be a general marker of the intensity of CE immune response and that could lead to a deeper understanding of the specific antigen-induced mechanisms underlying allergic reactions in the human host. PMID- 11472434 TI - Enhanced T cell apoptosis in common variable immunodeficiency: negative role of the fas/fasligand system and of the Bcl-2 family proteins and possible role of TNF-RS. AB - CVI is a primary immunodeficiency characterized by a failure of B cell differentiation associated with an array of T cell defects, such as enhanced T cell apoptosis. In this study we investigated the mechanisms underlying CVI enhanced T cell death. We analysed both the expression of Fas using flow cytometry techniques and the expression of FasL mRNA using RT-PCR in CVI T cells. We could not find any significant differences between CVI and normal subjects with regard to Fas expression, although there was a subgroup of CVI patients with very high Fas expression which was accompanied by an up-regulation of FasL mRNA. However, attempts to induce Fas-mediated apoptosis in these high Fas expressing cells, as evaluated by propidium iodide staining and APO2.7 staining, were unsuccessful. We also investigated intracellular levels of Bcl-2, bcl-xl and bax in CD4(+) and CD8(+) CVI T cells, as well as the bax/Bcl-2 ratio, using flow cytometry techniques but could not detect any differences between CVI and normal subjects. Finally we analysed TNF-RI and TNF-RII mRNA expression in CD4(+) and CD8(+) CVI T cells using semiquantitative RT-PCR and found a significant increase in expression of both TNF-Rs in CD4(+) T cells from CVI patients. Our data suggest that the increased expression of both TNF-Rs on T cells may be one of the mechanisms responsible for the accelerated T cell apoptosis in CVI. PMID- 11472435 TI - Initiation of alcoholic fatty liver and hepatic inflammation with a specific recall immune response in alcohol-consuming C57Bl/6 mice. AB - Whether immunological responses are involved in initiation and progression of alcoholic liver disease is unclear. We describe a mouse model of alcoholic liver injury characterized by steatosis and hepatic inflammation initiated by a recall immune response. Mice immune to Listeria monocytogenes fed a liquid diet containing ethanol and challenged with viable bacteria developed steatosis within 24 h and, at a later time, elevated serum alanine aminotransferase levels, indicating more liver damage in this group. Listeria antigen also induced steatosis and increased serum alanine aminotransferase levels in immune ethanol consuming mice. The production of tumour necrosis factor by a recall immune response in this model is a major, but not the only, component in initiation of alcoholic liver disease. PMID- 11472436 TI - Chemokines in human periodontal disease tissues. AB - An immunoperoxidase technique was used to examine IP-10 (interferon-gamma inducible protein 10), RANTES (regulated on activation normal T cell expressed and secreted), MCP-1 (monocyte chemoattractant protein-1), and MIP-1 alpha (macrophage inflammatory protein-1 alpha) in gingival biopsies from 21 healthy/gingivitis and 26 periodontitis subjects. The samples were placed into 3 groups according to the size of infiltrate. MIP-1 alpha+ cells were more abundant than the other chemokines with few MCP-1+ cells. The mean percent MIP-1 alpha+ cells was higher than the percent MCP-1+ cells (P = 0.02) in group 2 (intermediate size infiltrates) lesions from periodontitis subjects, other differences not being significant due to the large variations between tissue samples. Analysis of positive cells in relation to CD4/CD8 ratios showed that with an increased proportion of CD8+ cells, the mean percent MIP-1 alpha+ cells was significantly higher in comparison with the mean percent RANTES+ and MCP-1+ cells (P < 0.015). Endothelial cells were MCP-1+ although positive capillaries were found on the periphery of infiltrates only. Keratinocyte expression of chemokines was weak and while the numbers of healthy/gingivitis and periodontitis tissue sections positive for IP-10, RANTES and MCP-1 reduced with increasing inflammation, those positive for MIP-1 alpha remained constant for all groups. In conclusion, fewer leucocytes expressed MCP-1 in gingival tissue sections, however, the percent MIP-1 alpha+ cells was increased particularly in tissues with increased proportions of CD8 cells and B cells with increasing inflammation and also in tissues with higher numbers of macrophages with little inflammation. Further studies are required to determine the significance of MIP-1 alpha in periodontal disease. PMID- 11472437 TI - Impaired T cell proliferation, increased soluble death-inducing receptors and activation-induced T cell death in patients undergoing haemodialysis. AB - Haemodialysis is a widespread option for end-stage renal disease (ESRD). Long term success of dialysis is, however, limited by a high rate of serious bacterial and viral infections. We compared T cell functions in ESRD patients undergoing haemodialysis (n = 20), or were not dialysed and received conventional medical treatment (n = 20). Healthy volunteers (n = 15) served as controls. The T cell phenotype was examined by immunofluorescence using fluorochrome-labelled monoclonal antibodies and FACS analysis. The concentration of soluble CD95/Fas and of tumour necrosis factor-alpha receptor type 1 (sTNFR1) in the sera was quantified by ELISA. Activation-induced programmed T cell death was triggered by anti-CD3/CD28 antibodies and measured by 7-AAD staining. All immunological tests were performed at least 1 month after dialysis initiation. T cell proliferation in response to phytohaemagglutinin or anti-CD3 monoclonal antibodies was moderately diminished in non-dialysed patients and markedly reduced in haemodialysis patients compared to healthy controls (P < 0.01 and P < 0.001, respectively). In a mixed lymphocyte culture the proliferative response of T cells from dialysed patients was significantly diminished (P < 0.001). T cells of both non-dialysed and dialysed patients have augmented CD95/Fas and CD45RO expression, increased sCD95/Fas and sTNFR1 release and spontaneously undergo apoptosis. Culture of T cells from haemodialysis patients with anti-CD3/CD28 antibodies increased the proportion of CD4(+) T cells committing activation induced cell death by a mean 7.5-fold compared to T-helper cells from non dialysed patients (P < 0.001). Renal failure and initiation of haemodialysis results in a reduced proliferative T cell response, an aberrant state of T cell activation and heightened susceptibility of CD4(+) T cells to activation-induced cell death. PMID- 11472438 TI - Lupus anticoagulant and history of thrombosis are not associated with persistent endothelial cell activation in systemic lupus erythematosus. AB - Antiphospholipid antibodies (aPL), especially lupus anticoagulant (LAC), characterize systemic lupus erythematosus (SLE) patients at increased risk for arterial and venous thromboembolic complications. It has been reported that purified human anti-phospholipid antibodies cause endothelial cell activation in in vitro experiments. In order to investigate whether increased endothelial cell activation is associated with thromboembolic events in SLE patients with LAC, we measured plasma levels of thrombomodulin (TM), von Willebrand factor (vWf), sP selectin, vascular cell adhesion molecule-1 (sVCAM-1) and ED1-fibronectin in a study of 76 patients with SLE. Patients were subdivided on the basis of: no history of thrombosis and LAC-negative (n = 22) or LAC-positive (n = 17); positive history of thrombosis and LAC-negative (n = 16) or LAC-positive (n = 21). The median SLE disease activity index (SLEDAI) was 4. Although concentrations of sTM, vWf, sP-selectin and sVCAM-1 were significantly elevated in SLE compared with values in healthy controls, they did not differ between the four groups, between patients with or without history of thrombosis, and between patients with or without LAC. Presence of anticardiolipin antibodies could not explain these negative findings. Adjustment of the concentrations for significantly associated variables, such as age, hypertension, smoking, immunosuppressive treatment and concentrations of creatinine, cholesterol and homocysteine, did not change the main results of the study. Only sTM was significantly lower in patients with both LAC and thrombosis than in patients without both these features after adjustment for serum creatinine concentrations. In conclusion, we did not find an association between endothelial cell activation and presence of LAC or history of thrombosis in SLE. PMID- 11472439 TI - Selective recruitment of CCR6-expressing cells by increased production of MIP-3 alpha in rheumatoid arthritis. AB - Infiltration of various types of leucocytes has been shown to play a crucial role in the pathogenesis of rheumatoid arthritis (RA). Macrophage inflammatory protein 3 alpha (MIP-3 alpha) is a recently identified chemokine which is a selective chemoattractant for leucocytes such as memory T cells, naive B cells and immature dendritic cells. In this study, we investigated the expression of MIP-3 alpha and its specific receptor CCR6 in the inflamed joints of patients with RA. Increased amounts of MIP-3 alpha were found by ELISA in synovial fluids (SF) of patients with RA. MIP-3 alpha was apparently detected in all synovial tissue specimens of RA patients (n = 6), but it could not be detected in that of osteoarthritis (OA) patients (n = 4). Expression of MIP-3 alpha was detected especially in the sublining layer, and infiltrating mononuclear cells in RA synovial tissue. Gene expression of MIP-3 alpha was also found in six out of 11 RA-synovial fluid cells by RT-PCR. Cultured synovial fibroblasts derived from either RA or OA patients were capable of producing MIP-3 alpha in response to IL-1 beta and TNFalpha in vitro. Furthermore, expression of CCR6 was found in infiltrating mononuclear cells in the cellular clusters and around the vessels of RA synovial tissue. These findings indicate that increased production of MIP-3 alpha may contribute to the selective recruitment of CCR6-expressing cells in RA. PMID- 11472440 TI - Heterogeneity of human Ro ribonucleoproteins (RNPS): nuclear retention of Ro RNPS containing the human hY5 RNA in human and mouse cells. AB - Ro ribonucleoproteins (RNPs) are autoantigens that result from the association of a 60-kDa protein (Ro60) with a small RNA (hY1, hY3, hY4 or hY5 in humans, mY1 or mY3 in mice). Previous studies localized Ro RNPs to the cytoplasm. Because Ro RNPs containing hY5 RNA (Ro(hY5) RNPs) have unique biochemical and immunological properties, their intracellular localization was reassessed. Subcellular distribution of mouse and human Ro RNPs in intact and hY-RNA transfected cells was assessed by immunoprecipitation and Northern hybridization. Human Ro(hY1--4) RNPs as well as murine Ro(mY1, mY3) RNPs are exclusively cytoplasmic. Ro RNPs containing an intact hY5 RNA, but not those containing a mutated form of hY5 RNA, are found in the nuclear fractions of human and mouse cells. Ro(hY5) RNPs are stably associated with transcriptionally active La protein and are known to associate with RoBPI, a nuclear autoantigen. Our results demonstrate that Ro(hY5) RNPs are specifically present in the nucleus of cultured human and murine cells. The signal for nuclear localization of Ro(hY5) RNPs appears to reside within the hY5 sequence itself. In conclusion, we suggest that the unique localization and interactions of primate-specific Ro(hY5) RNPs reflect functions that are distinct from the predicted cytoplasmic function(s) of more conserved Ro RNPs. PMID- 11472441 TI - Different proteolytic mechanisms involved in Fc gamma RIIIb shedding from human neutrophils. AB - The Fc gamma receptor type IIIb (CD16) is highly expressed on human neutrophils and is found in a soluble form in plasma and in other body fluids. Upon activation of neutrophils in vitro, Fc gamma RIIIb is shed from the cell surface by proteolytic cleavage. We have now investigated the effect of metalloproteinase inhibitors and a serine proteinase inhibitor on the shedding of Fc gamma RIIIb induced by phorbol 12-myristate 13-acetate (PMA) or cytochalasin B (cyto B) + N formyl-methionyl-leucyl-phenylalanine (fMLP). Metalloproteinase inhibitors blocked to a large extent PMA-induced, but not cyto B + fMLP-induced shedding of Fc gamma RIIIb. Inhibition of members of the ADAM (a disintegrin and metalloproteinase) family appeared most efficient. In contrast, the serine protease inhibitor N-methoxysuccinyl-alanine-alanine-proline-valine chloromethylketone (MeOsuc-AAPV-CMK) largely blocked cyto B + fMLP-induced, but not PMA-induced shedding of Fc gamma RIIIb. Metalloproteinase inhibitors in combination with the serine proteinase inhibitor resulted in full inhibition of Fc gamma RIIIb shedding induced by either PMA or cyto B + fMLP. The shedding of Fc gamma RIIIb that accompanied apoptosis was inhibited by 60% in the presence of inhibitors of metalloproteinases but was insensitive to inhibition of serine proteinases. These results show that distinct types of proteolytic enzyme are involved in the stimulus-induced shedding of Fc gamma RIIIb from human neutrophils and suggest that these proteinases may become differentially activated under various physiological or pathological conditions. PMID- 11472442 TI - Predicting CHD risk in patients with diabetes mellitus. PMID- 11472443 TI - Management of obesity in patients with Type 2 diabetes. AB - Although prevention is clearly a logical first step in the management of the obese Type 2 diabetic patient, such programmes have had little long-term success. Diet, exercise and behavioural modification still form the cornerstones of treatment and relatively small weight loss results in improvement of all major obesity-related co-morbidities, including Type 2 diabetes. The obese diabetic patient faces extra impediments to weight loss, including the adverse effects of diabetic medication, poor glycaemic control and diabetes-related complications. New drugs may offer some additional help, in general by providing the benefit associated with the weight loss as such. Bariatric surgery can produce major long term weight loss in the severely obese. PMID- 11472444 TI - Coronary heart disease risk assessment in diabetes mellitus--a comparison of PROCAM and Framingham risk assessment functions. AB - AIMS: To assess any differences between coronary heart disease (CHD) risks calculated by the Framingham equation and those calculated by the PROCAM equation in men with and without diabetes mellitus, and whether any such differences are associated with the hypertriglyceridaemia of diabetes mellitus. METHODS: Clinical and biochemical data collected from 1774 men seen in either general practice, a hospital diabetes or lipid clinic. CHD risks were calculated by both the Framingham and PROCAM functions and comparisons made between those patients with and those without diabetes. RESULTS: Of the 1774 men only 996 fulfilled the criteria for assessment by the PROCAM equation and thus further analysis. Patients with diabetes mellitus had significantly higher serum triglyceride levels than those without (1.9 mmol/l vs. 1.7 mmol/l). Median annual CHD risks calculated by the Framingham function were 1.7% in the patients with and 1.32% in the patients without diabetes mellitus, whereas those calculated by the PROCAM function were 0.77% and 0.6%, respectively. Bland-Altman difference plots showed that in both groups of patients the PROCAM equation systematically underestimated risk in comparison with the Framingham equation at low levels of risk but overestimated at higher levels of risk. The shape of the plots in each group of patients was, however, similar. CONCLUSION: There were no systematic differences between CHD risks calculated by the two different equations in patients with diabetes compared with those without, despite the higher serum triglyceride levels associated with diabetes. Restrictions in the use of the PROCAM function meant that only 56% of the original cohort could be assessed in this way. Thus the Framingham equation remains the most suitable method of CHD risk prediction for UK patients with and without diabetes mellitus. PMID- 11472445 TI - Peritoneal dialysis: new developments and new problems. AB - AIMS: To ascertain the incidence of hyponatraemia, and the impact of an icodextrin-based dialysis solution regime on hyponatraemia, in diabetic and non diabetic patients using peritoneal dialysis (PD). METHODS: Following severe hyponatraemia, resulting in neurological sequelae, in two diabetic patients who were on icodextrin-based PD for established renal failure, we reviewed the results of all our PD patients. Data was gathered retrospectively, from our database and case notes, on plasma sodium, haematocrit, dry weight estimation, plasma albumin and residual renal function. Patients using icodextrin-based solutions were compared with a random selection of patients using dextrose-based peritoneal solutions. We also compared diabetic patients on and off icodextrin with their non-diabetic counterparts using the same dialysis regime. The data were analysed using the paired Student's t-test. RESULTS: Plasma sodium was significantly lower in all patients using icodextrin-based solutions compared with those patients on dextrose-based PD. Plasma sodium was also found to fall in all patients following the initiation of an icodextrin-based PD regime. The fall in plasma sodium was statistically significant in diabetic and non-diabetic patients, but only fell below the laboratory reference range in the diabetic patients. CONCLUSIONS: Icodextrin-based PD is a risk factor for hyponatraemia and may produce clinically relevant symptoms if, as in our two cases, the hyponatraemia is compounded by other factors. PMID- 11472446 TI - Child and parental mental ability and glycaemic control in children with Type 1 diabetes. AB - AIMS: Many psycho-social factors can affect the glycaemic control of children with Type 1 diabetes, but the influence of the intelligence of the child and their parents has not been reported. METHODS: Seventy-eight children and adolescents with Type 1 diabetes and their mothers performed standardized tests to assess psychometric intelligence. The children were aged (median (range)) 12.0 (5-17) years with duration of diabetes 5.0 (1.0-13.0) years and required an insulin dose of (mean +/- SD) 1.0 +/- 0.3 U/kg per day. The children completed the Wide Range Achievement Test 3 reading test (WRAT3) and Raven's Standard Progressive Matrices (RSPM). A mean annual HbA1c was calculated for each subject (8.6 +/- 1.4%). The mothers performed the National Adult Reading Test (NART) and provided details of the occupation of the main wage-earner in the family from which social class (SC) was derived. RESULTS: The HbA1c of the child correlated with their age (r = 0.26, P = 0.02), SC (Kendall's rank correlation, tau = 0.17, P = 0.03) and with the NART error score of their mother (r = 0.28, P = 0.01), but no correlation was observed with the child's WRAT3 or RSPM score. Stepwise regression revealed that age and NART error score were the strongest independent determinants of glycaemic control (total adjusted r2 = 0.117). CONCLUSIONS: Parental intelligence appears to have a significant influence on the glycaemic control of a child with Type 1 diabetes, accounting for 7.6% of the reliable variance in HbA1c. PMID- 11472447 TI - What is hypertension in diabetes? Ambulatory blood pressure in 137 normotensive and normoalbuminuric Type 1 diabetic patients. AB - AIMS: To establish reference data for ambulatory blood pressure (AMBP) in normotensive, normoalbuminuric Type 1 diabetic patients and characterize the relation to clinic blood pressure (BP). To evaluate the statement of the third working party of the British Hypertension Society (BHS) that a target clinic BP in diabetes < 140/80 corresponds to a target day-time AMBP < 130/75 mmHg. PATIENTS AND METHODS: AMBP were performed in 172 normoalbuminuric, adult Type 1 diabetic patients, who had never received anti-hypertensive drugs. Clinic BP was determined as the mean of at least three auscultatory (Hawskley random zero manometer) and as the mean of at least three oscillometric (Spacelabs) BP values obtained just prior to ambulatory monitoring. Five patients with more than three missing hours/24 h were excluded. RESULTS: For 30 patients auscultatory clinic BP exceeded 140 mmHg systolic and/or 90 mmHg diastolic. For the remaining 137 normotensive patients day-time AMBP was 125.7/77.2 mmHg and oscillometric clinic BP was 125.3/76.5 mmHg (mean difference 0.3/0.7 mmHg; 95% confidence interval (CI) -0.9 to 1.5/-0.3 to 1.7 mmHg, P = 0.6/P = 0.2). Sixty-five percent of the patients had a diastolic day-time AMBP > 75 mmHg. CONCLUSIONS: Clinic BP and day time AMBP measured by the same method were indistinguishable. The target for day time diastolic AMBP (< 75 mmHg) proposed by the BHS is too low and is based on the misconception that in normotensive subjects day-time AMBP is lower than clinic BP. If the BHS guidelines are strictly adhered to, the consequence may be overtreatment in patients with normoalbuminuria and no end organ damage. PMID- 11472448 TI - Augmentation of central arterial pressure in Type 2 diabetes. AB - AIMS: Aortic systolic blood pressure has been shown to be augmented in Type 1 diabetes, indicative of more rapid pulse wave reflection due to increased arterial stiffness. This abnormality is more pronounced in diabetic males. The aim of this study was to examine the effects of diabetes on augmentation of aortic systolic pressure in subjects with Type 2 diabetes. METHODS: Radial artery pressure waveforms were obtained non-invasively by applanation tonometry. A central aortic waveform can be derived using a transfer function obtained from previous studies during cardiac catheterization. A total of 88 subjects with Type 2 diabetes (51 men and 37 women, aged 55.8 years (interquartile range (IR) 49.7 64.1), duration of diabetes 7.5 years (IR 2.4-12.4), HbA1c 7.6% (IR 6.6-8.7)) and 85 controls subjects (40 men and 45 women, aged 55.3 years (IR 44.2-66.4)) were studied. The central aortic waveform allowed determination of the: (i) aortic augmentation index and (ii) subendocardial viability ratio. RESULTS: Similar to Type 1 diabetic subjects, patients with Type 2 diabetes had a significantly higher aortic augmentation index (136.1 +/- 18.0% vs. 128.3 +/- 19.2%, t = 2.8, P = 0.006) and lower subendocardial viability ratio (137.4 +/- 25.0% vs. 155.1 +/- 25.9%; t = 4.6, P = 0.0001) compared with controls. Multivariate analysis identified diabetes as an important determinant of aortic augmentation index (t = 4.0, P = 0.0001). The higher aortic augmentation index was due mainly to the male cohort (t = 2.6; P = 0.01) and was not apparent for females with diabetes (P = 0.2). CONCLUSIONS: Type 2 diabetes is characterized by higher augmentation of aortic systolic pressure and unfavourable ratio of myocardial perfusion to cardiac workload. These results are consistent with increased arterial stiffness. The age-related progression of arterial stiffness is similar in Type 1 and Type 2 diabetes. Anti-hypertensive agents that reduce wave reflection and augmentation may help to prevent systolic hypertension and cardiac hypertrophy in diabetes. PMID- 11472449 TI - Insulin secretion and insulin sensitivity are normal in non-diabetic subjects from maternal inheritance diabetes and deafness families. AB - BACKGROUND: The pathophysiological mechanism of diabetes mellitus in the presence of the 3243 A-G tRNALEU(UR) mitochondrial DNA mutation is thought to result from deficient insulin secretion. However, few subjects with normal glucose tolerance have been studied to determine the sequence of events resulting in the development of diabetes mellitus. AIM: To determine whether abnormalities of insulin sensitivity, insulin secretion or glucose effectiveness are present in non-diabetic subjects with the 3243 A-G tRNALEU(UUR) mitochondrial DNA mutation. METHODS: Twelve non-diabetic subjects with the mutation were compared with 12 controls, matched for age and anthropometric parameters, using both oral and intravenous glucose tolerance tests, the latter with Minimal Model analysis. RESULTS: Following an oral glucose load we found significantly higher blood glucose levels at 90 min and 120 min and significantly higher insulin levels at 120 min and 180 min in non-diabetic subjects with the mutation but no difference in the insulinogenic indices at 30 min and 180 min. From the intravenous glucose tolerance test there was no difference in overall glucose tolerance, insulin sensitivity, first- or second-phase insulin secretion, proinsulin secretion or glucose effectiveness. Insulin-independent glucose disposal was increased in subjects with lower insulin sensitivity and declined with increasing age in subjects with the mutation but not in controls. CONCLUSIONS: While there appear to be subtle defects of glucose handling in non-diabetic subjects with the 3243 mutation, these could not be explained by differences in insulin sensitivity or secretion. PMID- 11472450 TI - Association of macular involvement with proliferative retinopathy in Type 2 diabetes. AB - AIMS: To determine whether documented evidence of pre-existing maculopathy is present in Type 2 diabetic patients who require photocoagulation or vitrectomy for proliferative disease. This is a retrospective case notes review. METHODS: The study was performed at Queen's Medical Centre, UK. All patients listed in the laser register and operating theatre register over 5 years (March 1994 to March 1999) who had undergone pan retinal photocoagulation (PRP) for diabetic retinopathy, in one consultant firm, were studied. The medical records of patients who had undergone vitrectomy as their first treatment for proliferative diabetic retinopathy in a vitreo-retinal firm over the 5 years were examined. The proportion of patients with documented maculopathy before development of proliferative retinopathy was determined. RESULTS: All except 1/134 or 0.7% (95% confidence interval 0.1-4.5%) patients had documentation of macular signs in at least one eye prior to the patient's first session of PRP. Of the remaining 133, 104 (78%) had had focal or grid photocoagulation of the macula. The median time between the documentation of maculopathy before the first PRP treatment session was 16 months (interquartile range 7-36). Fourteen patients had vitrectomy and endolaser performed as the initial treatment for proliferative disease. All patients had documented maculopathy before the onset of proliferative disease. CONCLUSIONS: The results of this study suggest that in Type 2 diabetes, proliferative disease occurs relatively late compared with maculopathy. In such patients, maculopathy is almost invariably present when proliferative disease is detected. PMID- 11472451 TI - Comparison between repaglinide and glipizide in Type 2 diabetes mellitus: a 1 year multicentre study. AB - AIMS: To evaluate the long-term effectiveness and safety of repaglinide, a novel prandial glucose regulator, in comparison with glipizide in the treatment of patients with Type 2 diabetes. METHODS: Diet or tablet-treated patients with Type 2 diabetes (n = 256; age 40-75 years, body mass index (BMI) 20-35 kg/m2, HbA1c 4.2-12.8%), without signs of severe microvascular or macrovascular complications, were included in this double-blind, multicentre, parallel-group comparative trial. Patients were randomized at a 2:1 ratio to repaglinide, 1-4 mg at mealtimes, or glipizide, 5-15 mg daily. RESULTS: Changes in fasting blood glucose (FBG) and HbA1c during the 12 months of treatment showed a significant difference in favour of repaglinide. In oral hypoglycaemic agents (OHA)-naive patients, HbA1c decreased in the repaglinide and glipizide groups by 1.5% and 0.3%, respectively (P < 0.05 between groups). Fasting blood glucose decreased in the repaglinide group by 2.4 mmol/l and increased in the glipizide group by 1.0 mmol/l (P < 0.05 between groups). In the study population as a whole, repaglinide was able to maintain glycaemic control (HbA1c level) during the 1-year study period, whereas control deteriorated significantly with glipizide. Change in HbA1c from baseline was significantly better with repaglinide than with glipizide after 12 months (P < 0.05). In addition, FBG deteriorated significantly in the glipizide group compared with the repaglinide group (P < 0.05). No patients in either group experienced a major hypoglycaemic event; the number of patients experiencing minor hypoglycaemia was similar in the repaglinide and glipizide groups (15% and 19%, respectively). CONCLUSIONS: Repaglinide, given as a prandial glucose regulator, is shown to be an effective and safe treatment of patients with Type 2 diabetes, and is better than glipizide in controlling HbA1c and FBG levels, overall, and in OHA-naive patients. PMID- 11472452 TI - Longitudinal study of urinary albumin excretion in young diabetic patients- Wessex Diabetic Nephropathy Project. AB - AIMS: This study was established to follow changes in albumin/creatinine ratio (ACR) and to determine the prevalence and degree of progression of microalbuminuria (MA) or of clinical proteinuria (CP) in children with Type 1 diabetes. The study has investigated subjects for up to 12 years in establishing the correlation between MA and gender, age, duration of diabetes and glycated haemoglobin (HbA1c). The study has defined clinical cut-offs for MA in daytime clinic urine samples in young diabetic subjects. METHODS: Three hundred and sixty one patients were involved in the study, with 221 (61.2%) having over six sets of data. Urine samples were collected at routine annual clinic visits and analysed without prior freezing for ACR. Blood samples were taken for HbA1c measurement. Data including sex, age and duration of diabetes were recorded. RESULTS: A random clinic ACR of < 4.5 mg/mmol (males) and 5.2 mg/mmol (females) creatinine was used as the 'clinical cut-off' to define the presence of MA. The presence of MA was independent of HbA1c and duration of diabetes but appeared be associated with the adolescent years (> 10 years). There was little evidence of progression from normoalbuminuria to MA, or from MA to CP. Of patients aged 10-18 years, 30.9% of males and 40.4% of females had one or more episodes of MA. CONCLUSIONS: Persistent MA and random episodes of MA or CP may be associated with the adolescent years but not with duration of diabetes. Further study will reveal if the substantial increases in ACR sometimes seen during adolescence are predictive of diabetic nephropathy. Clinical cut-offs of < 4.5 and < 5.2 mg/mmol creatinine for males and females, respectively, are suggested for the interpretation of changes in ACR in random urine samples in young people with Type 1 diabetes. PMID- 11472453 TI - The National Paediatric Diabetes Register/Audit. PMID- 11472454 TI - Analysis of a polycytosine tract and heteroplasmic length variation in the mitochondrial DNA D-loop of patients with diabetes, MELAS syndrome and race matched controls. AB - AIM: The T to C substitution at position 16189 nt of the human mitochondrial genome has been associated with the development of heteroplasmic length variation in the control region of mtDNA. Previous reports have suggested that this defect may be associated with the development of other pathogenic mtDNA mutations, including the diabetogenic A to G mutation in the tRNALEU(UUR). Recently the 16189 nt variant has also been associated with insulin resistance in British adult men. In order to investigate these associations further we studied 23 patients with the 3243 nt mutation, 150 patients with Type 2 diabetes and 149 non diabetic controls. METHODS: The region around 16189 nt was investigated by polymerase chain reaction-restriction fragment length polymorphism analysis and automated sequencing. RESULTS: We find that the T to C substitution at 16189 nt is associated with heteroplasmic length variation only when the resultant polycytosine tract is not interrupted by a second mutation. There are no significant differences in the prevalence of the 16189 nt variant or heteroplasmic length variation between patients with the 3243 nt mutation, patients with Type 2 diabetes or race-matched normal controls. CONCLUSIONS: We conclude that these variants are likely to represent normal polymorphisms and that previously reported associations should be treated with caution unless they can be replicated in other populations. PMID- 11472455 TI - Predictive genetic testing in maturity-onset diabetes of the young (MODY). AB - INTRODUCTION: Maturity-onset diabetes of the young (MODY) is characterized by autosomal dominant inheritance of young-onset non-insulin-dependent diabetes. It accounts for approximately 1% of Type 2 diabetes (approximately 20 000 people in the UK). Diagnostic and predictive genetic tests are now possible for 80% of MODY families. Diagnostic tests can be helpful as the diagnosis can be confirmed and the subtype defined which has implications for treatment and prognosis. However predictive genetic testing, particularly in children, raises many scientific, ethical and practical questions. METHODS: This is a case report of a family with diabetes resulting from an hepatic nuclear factor (HNF)1alpha mutation, who request a predictive test in their 5-year-old daughter. The scientific issues arising from molecular genetic testing in MODY are discussed, along with the process of genetic counselling. The views of the family and the clinical genetics team involved are presented. RESULTS: The implications of positive and negative predictive test results and the possibility of postponing the test were among many issues discussed during genetic counselling. The family remained convinced the test was appropriate for their daughter and the clinical genetics team fully supported this decision. The family, motivated by their family history of diabetes and personal experiences of the disease, wished to reduce uncertainty about their daughter's future irrespective of the result. CONCLUSIONS: This case emphasizes that decisions on predictive testing are very personal and require appropriate counselling. PMID- 11472457 TI - Estimating the prevalence of diagnosed diabetes in a health district of Wales. PMID- 11472458 TI - Elevated matrix metalloproteinase-2 and -9 in urine, but not in serum, are markers of type 1 diabetic nephropathy. PMID- 11472459 TI - Silent hypoglycaemia at the diabetic clinic. PMID- 11472460 TI - Assessment of blood pressure in patients with Type 2 diabetes: comparison between home blood pressure monitoring, clinic blood pressure measurement and 24-h ambulatory blood pressure monitoring. AB - AIMS: To compare a home blood pressure (BP) monitoring device and clinic BP measurement with 24-h ambulatory BP monitoring in patients with Type 2 diabetes mellitus (DM). METHODS: Fifty-five patients with type 2 DM had BP measured at three consecutive visits to the DM clinic by nurses using a stethoscope and mercury sphygmomanometer (CBP). Twenty-four-hour ambulatory BP was measured using a Spacelabs 90207 automatic cuff-oscillometric device (ABPM). Subjects were then instructed in how to use a Boots HEM 732B semiautomatic cuff-oscillometric home BP monitoring device and measured BP at home on three specified occasions on each of 4 consecutive days at varying times (HBPM). RESULTS: Correlations between HBPM and ABPM were r = 0.88, P < 0.001 for systolic BP and r = 0.76, P < 0.001 for diastolic BP, with correlations between CBP and ABPM being systolic r = 0.59, P < 0.001, diastolic r = 0.47, P < 0.001. HBPM agreed with ABPM more closely compared with CBP (CBP +10.9/+3.8 (95% confidence intervals (CI) 6.9, 14.8/1.6, 6.1) vs. HBPM +8.2/+3.7 (95% CI 6.0, 10.3/2.0, 5.4)). The sensitivity, specificity and positive predictive value of HBPM in detecting hypertension were 100%, 79% and 90%, respectively, compared with CBP (85%, 46% and 58%, respectively). CONCLUSIONS: In patients with Type 2 DM, home BP monitoring is superior to clinic BP measurement, when compared with 24-h ambulatory BP, and allows better detection of hypertension. It would be a rational addition to the annual review process. Diabet. Med. 18, 431-437 (2001) PMID- 11472461 TI - An economic evaluation of atenolol vs. captopril in patients with Type 2 diabetes (UKPDS 54). AB - AIMS: To compare the net cost of a tight blood pressure control policy with an angiotensin converting enzyme inhibitor (captopril) or beta blocker (atenolol) in patients with Type 2 diabetes. DESIGN: A cost-effectiveness analysis based on outcomes and resources used in a randomized controlled trial and assumptions regarding the use of these therapies in a general practice setting. SETTING: Twenty United Kingdom Prospective Diabetes Study Hospital-based clinics in England, Scotland and Northern Ireland. SUBJECTS: Hypertensive patients (n = 758) with Type 2 diabetes (mean age 56 years, mean blood pressure 159/94 mmHg), 400 of whom were allocated to the angiotensin converting enzyme inhibitor captopril and 358 to the beta blocker atenolol. MAIN OUTCOME MEASURES: Life expectancy and mean cost per patient. RESULTS: There was no statistically significant difference in life expectancy between groups. The cost per patient over the trial period was 6485 UK pounds in the captopril group, compared with 5550 UK pounds in the atenolol group, an average cost difference of 935 UK pounds (95% confidence interval 188 UK pounds, 1682 UK pounds). This 14% reduction arose partly because of lower drug prices, and also because of significantly fewer and shorter hospitalizations in the atenolol group, and despite higher antidiabetic drug costs in the atenolol group. CONCLUSIONS: Treatment of hypertensive patients with Type 2 diabetes using atenolol or captopril was equally effective. However, total costs were significantly lower in the atenolol group. Diabet. Med. 18, 438-444 (2001) PMID- 11472462 TI - The apolipoprotein epsilon2 allele and the severity of coronary artery disease in Type 2 diabetic patients. AB - AIMS: To examine the hypothesis that apolipoprotein E2 is associated with more severe coronary disease in Type 2 diabetic patients. RESEARCH DESIGN AND METHODS: In this retrospective cohort study, 491 patients with angiographically assessed coronary disease were recruited from those attending a university hospital cardiology department. Participants completed detailed questionnaires, from which the presence or absence of diabetes was determined. Fasting blood samples were obtained for apolipoprotein E genotype and measurement of blood lipid parameters. RESULTS: The prevalence of triple vessel disease was significantly lower in non diabetic, epsilon2 allele carriers (39.3% vs. 16.2%; odds ratio (OR) 0.30 (0.12 0.71), P < 0.03) compared with E3/3 carriers. In Type 2 diabetic patients, epsilon2 allele carriers had an excess of triple vessel disease compared with E3/3 genotypes (43.3 vs. 68.8%; OR 2.8 (1.07-7.30), P < 0.05). The differences were independent of other variables. The apo E4 subgroup showed no significant differences in the frequency of triple vessel disease. CONCLUSIONS: Diabetic epsilon2 allele carriers had more severe coronary artery disease than diabetic patients with other apo E isoforms. In non-diabetic patients the epsilon2 allele appeared to protect against severe coronary disease. We hypothesize that interaction between the diabetic milieu and the epsilon2 allele accelerates plaque progression. It suggests that diabetic patients who are carriers of the epsilon2 allele, even in the heterozygous form, should be the focus of particular therapeutic attention. Diabet. Med. 18, 445-450 (2001) PMID- 11472463 TI - The relationship between active renin concentration and plasma renin activity in Type 1 diabetes. AB - AIMS: Circulating activity of the renin-angiotensin-aldosterone system (RAAS) can be assessed by measuring plasma active renin concentration (ARE), as well as by measuring plasma renin activity (PRA). We aimed to assess the relationships between ARE and PRA in Type 1 diabetic compared with non-diabetic control subjects. We also assessed concentrations of the active renin precursor, prorenin. PATIENTS AND METHODS: Thirty-five Type 1 diabetic subjects and 34 non diabetic control subjects were assessed. Groups had similar ages, sex distributions, body mass indices, systolic and diastolic blood pressures. PRA was measured by radioimmunoassay of angiotensin I generation from endogenous substrate. ARE and total renin concentration (TRE) were measured by immunoradiometric assay (Nichols Institute Diagnostics, USA). Prorenin concentration was calculated as the difference between ARE and TRE. RESULTS: PRA was significantly lower in Type 1 diabetic than in control subjects (0.8 (0.4 1.1) vs. 1.1 (0.9-1.9) pmol/ml per h; P < 0.005), while ARE was similar (17 (9 33) vs. 18 (15-25) mU/l; P = 0.548). PRA (loge transformed) correlated strongly with ARE in diabetic (r = 0.49; P = 0.003) and control subjects (r = 0.59; P = 0.0002), but there was significant vertical separation of the regression lines for the two groups (P < 0.0001). Prorenin concentrations were significantly higher in Type 1 diabetic subjects (249 (170-339) vs. 171 (153-219) mU/l; P = 0.005). Diabetic subjects with high prorenin concentrations (> 400 mU/l (control mean + 3 SD)) were more likely to have microalbuminuria (P = 0.027) and peripheral neuropathy (P = 0.049). CONCLUSIONS: Type 1 diabetes is associated with an altered relationship between ARE and PRA, such that ARE is higher for a given PRA compared with non-diabetic control subjects. Both ARE and PRA are used to assess circulating RAAS activity. The altered relationship between the two in Type 1 diabetic subjects suggests that neither parameter alone is necessarily an adequate and reliable index of such activity. Higher prorenin concentrations, particularly in association with microvascular complications, were confirmed in the Type 1 diabetic subjects. Diabet. Med. 18, 451-458 (2001) PMID- 11472464 TI - The Utrecht Diabetes Project: telemedicine support improves GP care in Type 2 diabetes. AB - The Utrecht Diabetes Project (UDP) is a shared-care project providing remote diabetologist support for 85 GPs. In the UDP all examinations, performed by the GP, follow standardized procedures, results being sent to the diabetologist. Laboratory results are sent automatically to both GP and diabetologist. AIMS: To study the composition of the UDP population; completeness of data recording; changes in biochemical variables of UDP patients; and GPs' motives for enrolling them. METHODS: Data were extracted from the records of 19 GPs and a questionnaire was sent to all UDP GPs. RESULTS: Of 770 patients with Type 2 diabetes, 44% were treated with UDP support, 29% by their GPs alone, and 27% at out-patient clinics. The 336 UDP patients were representative of all UDP patients. Patients older than 75 years were the greater part of those treated by GPs alone; out-patient clinics had more patients with diabetes > 10 years or with complications. UDP patients' records were the most complete. Diabetes regulation, lipid levels and diastolic blood pressure in the UDP patients improved significantly after inclusion in the UDP. GPs tend to seek UDP aid especially for patients who are young, or of recent onset, at risk of macrovascular complications, or needing insulin. CONCLUSIONS: Standardized data transfer between GP, diabetologist and laboratory might establish an effective infrastructure for shared diabetes care. Diabet. Med. 18, 459-463 (2001) PMID- 11472465 TI - Variability of glomerular filtration rate estimation using single injection methods in Type 1 diabetic patients. AB - AIM: Single-injection inulin clearance (SIIC) methods without urine collection have often been compared with classical constant-infusion inulin clearance methods. Therefore, we used repeated SIIC measurements to prove the hypothesis of a possible overestimation or variability of inulin clearance in Type 1 diabetic patients. METHODS: Two SIIC periods, glomerular filtration rate 1 and 2 (GFR 1, GFR 2) were performed consecutively on the same day. In diabetic patients, GFR 3 was measured several days later at the same time of day as the GFR 1 had been done. The calculation used a two-compartment modelling system (TCM) which showed excellent agreement to ratio of dose over area under the curve (D/AUC) calculation. RESULTS: Twelve normotensive Type 1 diabetic patients (mean and SD), age 39 (10.7) years, diabetes duration 20 (10.7) years, HbA1c 8.5% (0.82%), albuminuria 13.8 (15.7) mg/24 h and six controls, age 26 (2.6) years were examined. Healthy controls showed an excellent repeatability between GFR 1 111.8 (11.44) and GFR 2 110.8 (11.41) ml.min-1.1.73 m2-1. However, in diabetic patients there was a highly significant decrease between GFR 1 129.4 (11.86) and GFR 2 118.1 (13.05) ml.min-1.1.73 m2-1. GFR 1 with 129.4 (11.86) ml.min-1.1.73 m2-1 did not differ significantly from GFR 3 with 129.9 (12.40) ml.min-1.1.73 m2-1 in diabetic patients (P < 0.839). CONCLUSIONS: In contrast to normal subjects, in Type 1 diabetic patients single-injection inulin clearance technique showed a clinically relevant decrease in GFR on two repeated measurements on the same day. Overestimation of GFR by the first inulin clearance may be caused by incomplete inulin distribution in the slow compartment. Diabet. Med. 18, 464-468 (2001) PMID- 11472466 TI - Vibration perception threshold testing in patients with diabetic neuropathy: ceiling effects and reliability. AB - AIMS: To test the reliability of a new vibrometer (Maxivibrometer) which was constructed so that vibration perception threshold (VPT) could be determined without the disadvantage of the off-scale measurements frequently experienced with the Biothesiometer. METHODS: The two devices were compared and tested on a group of diabetic neuropathic subjects and a group of healthy, matched control subjects. VPT was tested on the plantar surface of the feet. RESULTS: The Maxivibrometer gave an actual measurement in all cases even if subjects were severely neuropathic. The replication-to-replication and day-to-day intraclass correlation coefficients for the Maxivibrometer VPT were, except in one case, above 0.94, indicating excellent reliability. The Biothesiometer VPT could also be measured with excellent reliability but only within a limited range of mild to moderate neuropathy, so it appears to be an appropriate screening tool. The replication-to-replication intraclass correlation coefficient was 0.93. CONCLUSIONS: Because VPT could be measured over a wide range with the Maxivibrometer, it was demonstrated that loss of sensation in diabetic neuropathy can progress far beyond the maximum VPT value of the Biothesiometer. The wide measurement range and the excellent reliability make the Maxivibrometer a valuable research tool to quantify loss of sensation, particularly in the presence of severe neuropathy and to record changes over time. Diabet. Med. 18, 469-475 (2001) PMID- 11472467 TI - Insulin resistance as a major determinant of increased coronary heart disease risk in postmenopausal women with Type 2 diabetes mellitus. AB - AIM: To investigate the risk factors associated with clinically defined coronary heart disease (CHD) in women with Type 2 diabetes mellitus (DM). METHODS: CHD status was assessed via standard history and resting electrocardiogram in 41 postmenopausal diabetic and 41 age- and body mass index-matched normoglycaemic women recruited from a community-based cohort. The following parameters were assessed: body composition by dual energy X-ray absorptiometry, blood pressure, metabolic and lipoprotein profile and haemostatic factors. RESULTS: Diabetic women with CHD (n = 14) had greater insulin resistance, calculated by homeostasis model assessment (10.2 (7.0-14.8) vs. 6.5 (5.5-7.7), P = 0.010), and higher plasminogen activator inhibitor-1 (PAI-1) levels (45 (29-69) vs. 24 (19-32) ng/ml, P = 0.013), than those without CHD. They also had higher triglycerides (2.9 (2.2-3.8) vs. 2.1 (1.8-2.4) mmol/l, P = 0.016) and a trend towards reduced low-density lipoprotein particle size (25.5 +/- 0.6 vs. 25.8 +/- 0.5 nm, P = 0.097). In a logistic regression model, insulin resistance was a significant independent predictor of CHD status (odds ratio = 1.33, 95% confidence interval = 1.06-1.68, P = 0.015). In contrast, in normoglycaemic women the major risk factors for CHD were elevated cholesterol, apolipoprotein(a), apolipoprotein B and systolic blood pressure (P = 0.018, P = 0.016, P = 0.006 and P = 0.049, respectively). CONCLUSIONS: Increased insulin resistance in association with elevated PAI-1 and dyslipidaemia appears to underpin the increased risk of CHD in women with Type 2 DM. Therapeutic approaches that increase insulin sensitivity may serve to reduce CHD risk in this vulnerable group. Diabet. Med. 18, 476-482 (2001) PMID- 11472468 TI - Contraindications to metformin therapy in patients with Type 2 diabetes--a population-based study of adherence to prescribing guidelines. AB - AIMS: To define the number of people in Tayside, Scotland (population 349 303) with Type 2 diabetes who use metformin, the incidence of contraindications to its continued use in these people and the proportion that discontinued metformin treatment following the development of a contraindication. METHODS: Retrospective cohort study of the incidence of contraindications to metformin in all patients with Type 2 diabetes using metformin from January 1993 to June 1995. The contraindications of acute myocardial infarction, cardiac failure, renal impairment and chronic liver disease were identified by: the regional diabetes information system, biochemistry database and hospital admissions database and a database of all encashed community prescriptions. RESULTS: One thousand eight hundred and forty seven subjects (26.3% of those with Type 2 diabetes) redeemed prescriptions for metformin. Of these, 3.5% were admitted with an acute myocardial infarction (71 episodes); 4.2% were admitted with cardiac failure (114 episodes); 21.0% received metformin and loop diuretics for cardiac failure concurrently; 4.8% developed renal impairment; and 2.8% developed chronic liver disease. The development of contraindications rarely resulted in discontinuation of metformin, for example only 17.5% and 25% stopped metformin after admission with acute myocardial infarction and development of renal impairment, respectively. In total, 24.5% of subjects receiving metformin, 6.4% of all people with Type 2 diabetes, had contraindications to its use. There was one episode of lactic acidosis in 4600 patient years. CONCLUSIONS: This population-based study shows that 24.5% of patients prescribed metformin have contraindications to its use. Development of contraindications rarely results in discontinuation of metformin therapy. Despite this, lactic acidosis remains rare. Diabet. Med. 18, 483-488 (2001) PMID- 11472469 TI - The influence of isolated small nerve fibre dysfunction on microvascular control in patients with diabetes mellitus. AB - AIM: The aim of the study was to investigate the influence of isolated small nerve fibre dysfunction on microvascular skin blood flow and transcutaneous oxygen tension in patients with diabetes mellitus. METHODS: Small nerve fibre dysfunction was assessed by the measurement of thermal and pain perception thresholds. Patients with evidence of large fibre disturbances as evaluated by means of vibration perception threshold were excluded from the study. Microvascular blood flow was investigated with laser-Doppler-fluxmetry (LDF) following stimulation with acetylcholine and mild thermal injury. RESULTS: Diabetic patients with small nerve fibre injury showed a significantly reduced increase in the laser-Doppler-flux signal following the application of acetylcholine compared with patients without neuropathy or healthy control subjects (2.8 arbitrary units (AU) (1.3-5.5) vs. 7.2 AU (4.1-25.5); P = 0.007 and vs. 8.5 AU (3.0-17.0), P = 0.02, respectively). The increase in LDF following thermal injury was also diminished in patients with small nerve fibre dysfunction compared with patients without neuropathy or the control group (29.8 AU (17.2 46.5) vs. 51.2 AU (29.5-93.5); P = 0.02 and vs. 54.6 AU (39.7-97.7); P = 0.004, respectively). In addition, they showed a significantly reduced transcutaneous oxygen tension compared with the other groups (42.9 mmHg (41.6-55.5) vs. 56.1 mmHg (49.2-60.8); P = 0.04 and vs. 59.0 mmHg (54.6-80.3), P = 0.03, respectively). CONCLUSIONS: Our study confirms an association between small nerve fibre injury and skin microvascular dysfunction. It further underlines the concept of neurovascular disturbances in the pathogenesis of neurotrophic foot ulceration. Diabet. Med. 18, 489-494 (2001) PMID- 11472470 TI - Calcaneal ultrasonometry in patients with Charcot osteoarthropathy and its relationship with densitometry in the lumbar spine and femoral neck and with markers of bone turnover. AB - AIMS: To assess calcaneal ultrasonometry in Charcot osteoarthropathy (CO) and to compare it with densitometry measured by dual energy X-ray absorptiometry (DEXA) and with bone remodelling markers. PATIENTS AND METHODS: A group of 16 diabetic patients in the acute stage of CO with a mean age (+/- SD) of 51 +/- 13 years was compared with 26 sex- and age-matched control subjects. Both calcaneal quantitative ultrasound (QUS) parameter stiffness and bone mineral density (BMD) measured in lumbar spine and femoral neck by DEXA were compared. Collagen type I cross-linked C-telopeptides (ICTP) were used for assessment of bone resorption. RESULTS: Patients with acute CO had significantly lower stiffness of the calcaneus in the Charcot and non-Charcot foot (both P < 0.001) and significantly lower femoral neck BMD (P < 0.05) in comparison with the control group. The T score of stiffness was significantly lower in the Charcot foot compared with the non-Charcot foot (-3.00 +/- 1.39 vs. -2.36 +/- 1.12; P < 0.01) and significantly lower than the mean T-score of BMD in the lumbar spine (-0.57 +/- 1.28; P < 0.001) and femoral neck (-1.58 +/- 1.24; P < 0.05). A significant difference in ICTP (8.49 +/- 4.37 vs. 3.92 +/- 2.55 ng/ml; P < 0.001) between patients with CO and the control group was found, and a significant correlation was demonstrated between ICTP and the T-score of stiffness (r = -0.73; P < 0.01). CONCLUSION: The lower calcaneal QUS parameter stiffness in the Charcot foot in comparison with the control group, with the non-Charcot foot and with BMD in the lumbar spine and femoral neck, and its association with increased bone resorption indicate that calcaneal ultrasonometry may be useful in diagnosing the acute stage of CO and in assessing the risk of foot fracture. Diabet. Med. 18, 495-500 (2001) PMID- 11472471 TI - Ethnicity and glycaemic control are major determinants of diabetic dyslipidaemia in Malaysia. AB - AIMS: To define the prevalence of dyslipidaemia in young diabetic patients in Peninsular Malaysia and the contributory factors of dyslipidaemia in these subjects. METHODS: This is a cross-sectional study involving 848 young diabetic patients from seven different centres, with representation from the three main ethnic groups. Clinical history and physical examination was done and blood taken for HbA1c, fasting glucose, total cholesterol, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol and triglycerides. RESULTS: The overall lipids were suboptimal, worse in Type 2 diabetes mellitus (DM) patients compared with Type 1 DM patients. Of the Type 2 patients, 73.2% had total cholesterol > 5.20 mmol/l, 90.9% had LDL-cholesterol > 2.60 mmol/l, 52.6% had HDL cholesterol < 1.15 mmol/l and 27.3% had serum triglycerides > 2.30 mmol/l. There were ethnic differences in the lipid levels with the Malays having the highest total cholesterol (mean 6.19 mmol/l), and the highest LDL-cholesterol (mean 4.16 mmol/l), while the Chinese had the highest HDL-cholesterol (geometric mean 1.24 mmol/l). Ethnicity was an important determinant of total, LDL- and HDL cholesterol in Type 2 DM, and LDL- and HDL-cholesterol and triglycerides in Type 1 DM. Glycaemic control was an important determinant of total, LDL-cholesterol and triglycerides in both Type 1 and Type 2 DM. Waist-hip ratio (WHR) was an important determinant of HDL-cholesterol and triglycerides in both types of DM. Gender was an important determinant of HDL-cholesterol in Type 2 DM, but not in Type 1 DM. Socioeconomic factors and diabetes care facilities did not have any effect on the dyslipidaemia. CONCLUSIONS: The prevalence of dyslipidaemia was high especially in Type 2 DM patients. Ethnicity, glycaemic control, WHR, and gender were important determinants of dyslipidaemia in young diabetic patients. Diabet. Med. 18, 501-508 (2001) PMID- 11472472 TI - Short-term Acipimox decreases the ability of plasma from Type 2 diabetic patients and healthy subjects to stimulate cellular cholesterol efflux: a potentially adverse effect on reverse cholesterol transport. AB - AIMS: To evaluate the effect of short-term administration of the anti-lipolytic agent, Acipimox, on the ability of plasma to stimulate cellular cholesterol removal, which represents one of the first steps in the anti-atherogenic process of reverse cholesterol transport. METHODS: Eight male Type 2 diabetic patients and eight healthy subjects were studied after a 12-h fast at baseline, after 24 h of Acipimox administration, 250 mg every 4 h, and again after 1 week (recovery). Plasma lipids, apolipoprotein AI, phospholipid transfer protein (PLTP) activity, pre-beta high-density lipoproteins (HDL) in incubated plasma and efflux of radiolabelled cholesterol from Fu5AH rat hepatoma cells to plasma were measured at each time point. RESULTS: Acipimox lowered plasma triglycerides in diabetic patients (P = 0.001) and healthy subjects (P = 0.002), whereas plasma non esterified fatty acids were decreased in diabetic patients (P = 0.001) compared with the averaged values at baseline and recovery. Acipimox decreased HDL cholesterol in healthy subjects (P = 0.007) and plasma apolipoprotein AI in both groups (P = 0.001 for diabetic patients; P = 0.008 for healthy subjects). Not only plasma PLTP activity (P = 0.001 for diabetic patients; P = 0.01 for healthy subjects), but also pre-beta HDL in incubated plasma (P = 0.001 for diabetic patients; P = 0.03 for healthy subjects) and cellular cholesterol efflux to plasma (P = 0.04 for diabetic patients; P = 0.005 for healthy subjects) were lowered by Acipimox in both groups. CONCLUSIONS: Short-term Acipimox administration impairs the ability of plasma from Type 2 diabetic patients and healthy subjects to stimulate cellular cholesterol efflux, in conjunction with alterations in HDL parameters and in PLTP activity. If the impairment of cellular cholesterol efflux to plasma is sustained with long-term treatment, this potentially adverse effect should be considered when treating diabetic dyslipidaemia with Acipimox. Diabet. Med. 18, 509-513 (2001) PMID- 11472473 TI - Coincidence of hand and foot ulceration in people with diabetes. PMID- 11472474 TI - Localized insulin allergy treated with continuous subcutaneous insulin. PMID- 11472500 TI - Functional and structural response of a cellulose-degrading methanogenic microbial community to multiple aeration stress at two different temperatures. AB - Two cellulose-fermenting methanogenic enrichment cultures originating from rice soil, one at 15 degrees C with Methanosaeta and the other at 30 degrees C with Methanosarcina as the dominant acetoclastic methanogen, both degraded cellulose anaerobically via propionate, acetate and H2 to CH4. The degradation was a two stage process, with CH4 production mainly from H2/CO2 and accumulation of acetate and propionate during the first, and methanogenic consumption of acetate during the second stage. Aeration stress of 12, 24, 36 and 76 h duration was applied to these microbial communities during both stages of cellulose degradation. The longer the aeration stress, the stronger the inhibition of CH4 production at both 30 degrees C and 15 degrees C. The 72 h stressed culture at 30 degrees C did not fully recover. Aeration stress at 30 degrees C exerted a more pronounced effect, but lasted for a shorter time than that at 15 degrees C. The aeration stress was especially effective during the second stage of fermentation, when consumption of acetate (and to a lesser extent propionate) was also increasingly inhibited as the duration of the stress increased. The patterns of CH4 production and metabolite accumulation were consistent with changes observed in the methanogenic archaeal community structure. Fluorescence in situ hybridization showed that the total microbial community at the beginning consisted of about 4% and 10% archaea, which increased to about 50% and 30% during the second stage of cellulose degradation at 30 degrees C and 15 degrees C respectively. Methanosarcina and Methanosaeta species became the dominant archaea at 30 degrees C and 15 degrees C respectively. The first round of aeration stress mainly reduced the non Methanosarcina archaea (30 degrees C) and the non-Methanosaeta archaea (15 degrees C). Aeration stress also retarded the growth of Methanosarcina and Methanosaeta at 30 degrees C and 15 degrees C respectively. The longer the stress, the lower was the percentage of Methanosarcina cells to total microbial cells after the first stress at 30 degrees C. A later aeration stress decreased the population of Methanosarcina (at 30 degrees C) in relation to the duration of stress, so that non-Methanosarcina archaea became dominant. Hence, aeration stress affected the acetotrophic methanogens more than the hydrogenotrophic ones, thus explaining the metabolism of the intermediates of cellulose degradation under the different incubation conditions. PMID- 11472501 TI - Direct amplification of nodD from community DNA reveals the genetic diversity of Rhizobium leguminosarum in soil. AB - Sequences of nodD, a gene found only in rhizobia, were amplified from total community DNA isolated from a pasture soil. The polymerase chain reaction (PCR) primers used, Y5 and Y6, match nodD from Rhizobium leguminosarum biovar trifolii, R. leguminosarum biovar viciae and Sinorhizobium meliloti. The PCR product was cloned and yielded 68 clones that were identified by restriction pattern as derived from biovar trifolii [11 restriction fragment length polymorphism (RFLP) types] and 15 clones identified as viciae (seven RFLP types). These identifications were confirmed by sequencing. There were no clones related to S. meliloti nodD. For comparison, 122 strains were isolated from nodules of white clover (Trifolium repens) growing at the field site, and 134 from nodules on trap plants of T. repens inoculated with the soil. The nodule isolates were of four nodD RFLP types, with 77% being of a single type. All four of these patterns were also found among the clones from soil DNA, and the same type was the most abundant, although it made up only 34% of the trifolii-like clones. We conclude that clover selects specific genotypes from the available soil population, and that R. leguminosarum biovar trifolii was approximately five times more abundant than biovar viciae in this pasture soil, whereas S. meliloti was rare. PMID- 11472502 TI - Development of specific oligonucleotide probes for the identification and in situ detection of hydrocarbon-degrading Alcanivorax strains. AB - The genus Alcanivorax comprises diverse hydrocarbon-degrading marine bacteria. Novel 16S rRNA-targeted oligonucleotide DNA probes (ALV735 and ALV735-b) were developed to quantify two subgroups of the Alcanivorax/Fundibacter group by fluorescence in situ hybridization (FISH), and the conditions for the single mismatch discrimination of the probes were optimized. The specificity of the probes was improved further using a singly mismatched oligonucleotide as a competitor. The growth of Alcanivorax cells in crude oil-contaminated sea water under the biostimulation condition was investigated by FISH with the probe ALV735, which targeted the main cluster of the Alcanivorax/Fundibacter group. The size of the Alcanivorax population increased with increasing incubation time and accounted for 91% of the 4',6-diamidino-2-phenylindole (DAPI) count after incubation for 2 weeks. The probes developed in this study are useful for detecting Alcanivorax populations in petroleum hydrocarbon-degrading microbial consortia. PMID- 11472503 TI - Bacterial community associated with Pfiesteria-like dinoflagellate cultures. AB - Dinoflagellates (Eukaryota; Alveolata; Dinophyceae) are single-cell eukaryotic microorganisms implicated in many toxic outbreaks in the marine and estuarine environment. Co-existing with dinoflagellate communities are bacterial assemblages that undergo changes in species composition, compete for nutrients and produce bioactive compounds, including toxins. As part of an investigation to understand the role of the bacteria in dinoflagellate physiology and toxigenesis, we have characterized the bacterial community associated with laboratory cultures of four 'Pfiesteria-like' dinoflagellates isolated from 1997 fish killing events in Chesapeake Bay. A polymerase chain reaction with oligonucleotide primers specific to prokaryotic 16S rDNA gene sequences was used to characterize the total bacterial population, including culturable and non-culturable species, as well as possible endosymbiotic bacteria. The results indicate a diverse group of over 30 bacteria species co-existing in the dinoflagellate cultures. The broad phylogenetic types of dinoflagellate-associated bacteria were generally similar, although not identical, to those bacterial types found in association with other harmful algal species. Dinoflagellates were made axenic, and the culturable bacteria were added back to determine the contribution of the bacteria to dinoflagellate growth. Confocal scanning laser fluorescence microscopy with 16S rDNA probes was used to demonstrate a physical association of a subset of the bacteria and the dinoflagellate cells. These data point to a key component in the bacterial community being species in the marine alpha-proteobacteria group, most closely associated with the alpha-3 or SAR83 cluster. PMID- 11472504 TI - Use of differential fluorescence induction and optical trapping to isolate environmentally induced genes. AB - The techniques of differential fluorescence induction (DFI) and optical trapping (OT) have been combined to allow the identification of environmentally induced genes in single bacterial cells. Designated DFI-OT, this technique allows the in situ isolation of genes driving the expression of green fluorescent protein (Gfp) using temporal and spatial criteria. A series of plasmid-based promoter probe vectors (pOT) was developed for the construction of random genomic libraries that are linked to gfpUV or egfp. Bacteria that do not express Gfp on laboratory medium (i.e. non-fluorescent) were inoculated into the environment, and induced genes were detected with a combined fluorescence/optical trapping microscope. Using this selection strategy, rhizosphere-induced genes with homology to thiamine pyrophosphorylase (thiE) and cyclic glucan synthase (ndvB) were isolated. Other genes were expressed late in the stationary phase or as a consequence of surface-dependent growth, including fixND and metX, and a putative ABC transporter of putrescine. This strategy provides a unique ability to combine spatial, temporal and physical information to identify environmental regulation of bacterial gene expression. PMID- 11472505 TI - Methyl tert-butyl ether (MTBE) degradation by a microbial consortium. AB - The widespread use of methyl tert-butyl ether (MTBE) as a gasoline additive has resulted in a large number of cases of groundwater contamination. Bioremediation is often proposed as the most promising alternative after treatment. However, MTBE biodegradation appears to be quite different from the biodegradation of usual gasoline contaminants such as benzene, toluene, ethyl benzene and xylene (BTEX). In the present paper, the characteristics of a consortium degrading MTBE in liquid cultures are presented and discussed. MTBE degradation rate was fast and followed zero order kinetics when added at 100 mg l(-1). The residual MTBE concentration in batch degradation experiments ranged from below the detection limit (1 microg l(-1)) to 50 microg l(-1). The specific activity of the consortium ranged from 7 to 52 mgMTBE g(dw)(-1) h(-1) (i.e. 19-141 mgCOD g(dw) ( 1) h(-1)). Radioisotope experiments showed that 79% of the carbon-MTBE was converted to carbon-carbon dioxide. The consortium was also capable of degrading a variety of hydrocarbons, including tert-butyl alcohol (TBA), tert-amyl methyl ether (TAME) and gasoline constituents such as benzene, toluene, ethylbenzene and xylene (BTEX). The consortium was also characterized by a very slow growth rate (0.1 d(-1)), a low overall biomass yield (0.11 gdw g(-1)MTBE; i.e. 0.040 gdw gCOD(-1)), a high affinity for MTBE and a low affinity for oxygen, which may be a reason for the slow or absence of MTBE biodegradation in situ. Still, the results presented here show promising perspectives for engineering the in situ bioremediation of MTBE. PMID- 11472507 TI - The modern matron: reborn or recycled? PMID- 11472508 TI - Leadership behaviours: effects on job satisfaction, productivity and organizational commitment. AB - BACKGROUND: Research in the west has shown that job satisfaction, productivity and organizational commitment are affected by leadership behaviours. The purpose of this study is to determine the effect of leadership behaviours on employee outcomes in Singapore. Very little research related to this subject has been done in health care settings in this country. The comparison of the results of the different types of settings and samples will allow a better understanding of the relationship between leadership behaviours and employee outcomes and thus help to determine if leadership is worth the extra effort. METHOD: The study explored the relationships between five leadership behaviours identified by Kouzes and Posner and the employee outcomes of registered nurses practising in the general wards, intensive care units and the coronary care unit in an acute hospital. Survey questionnaires were used to elicit responses from 100 registered nurses and 20 managers belonging to the organization. Data collected included demographic characteristics and the degree to which the five types of leadership behaviours were used as perceived by the nurse managers and the registered nurses. In addition, the level of nurse job satisfaction, the degree of productivity and the extent of organizational commitment are described. FINDINGS: The findings show a similar trend to the original studies in the United States of America. Use of leadership behaviours and employee outcomes were significantly correlated. The regression results indicate that 29% of job satisfaction, 22% of organizational commitment and 9% of productivity were explained by the use of leadership behaviours. Recommendations are made in the light of these findings. PMID- 11472509 TI - Comparing and contrasting the clinical nurse specialist and the advanced nurse practitioner roles. AB - There is currently a great deal of ambiguity regarding the difference between the role of clinical nurse specialist and advanced nurse practitioner. In distinguishing one title from another, factors such as the educational requirements of such, what the role involves, who the client is and whether the role encroaches on a doctor's role are discussed. This paper sets out the factors that are seen by some as significant in distinguishing between a clinical nurse specialist and advanced nurse practitioner. PMID- 11472510 TI - The role of the infection control nurse as a clinical nurse specialist or advanced nurse practitioner. AB - The specialty of infection control is examined regarding the role of the infection control nurse and the educational requirements for practitioners in this field in relation to post titles of clinical nurse specialist and advanced nurse practitioner. Whilst not fulfilling all the role constituents of a clinical nurse specialist, the infection control nurse is seen as more akin to a clinical nurse specialist than an advanced nurse practitioner. PMID- 11472511 TI - From competition to collaboration in the delivery of health care: England and Scotland compared. AB - AIM: The purpose of this paper is to outline and critically appraise the NHS reforms that were operationalized in April 1999. DISCUSSION: The paper recognizes that there are important differences between England and Scotland in the design and implementation of change. Differences are discernible in terms of the organization of the commissioning role and of the acute hospital sector, but more significant differences are evident in terms of the nature and organization of primary care. CONCLUSIONS: The paper concludes by indicating that a major challenge exists in the change management agenda with reference to primary care. Thus, while differences exist both north and south of the border, a shared feature is that the change management agenda in both England and Scotland is formidable. The main element of this challenge is that GPs have, in different ways, had to be drawn into the mainstream of the NHS. PMID- 11472512 TI - Consumerism in health care: the case of medication education. AB - BACKGROUND: The United Kingdom government's policy documents spanning the last decade clearly envisage the patient as a consumer of health care. In this context this paper discusses recent research findings related to the health-promotion practice of medication delivered by nurses in England in a variety of health care settings. Literature exploring consumerism in health care highlights a number of principles which were used to develop a framework to evaluate the data collected in this study. METHOD: Non-participant observation and audio-recordings of nurse patient interactions about medications were collected in seven different contexts focusing on adults, older people, mental health and community nurse settings. Post-interaction interviews with nurse and patient participants were conducted to explore views on quality, satisfaction with, and intended outcomes of, the interactions. FINDINGS: Generally, the findings demonstrated that the espoused theory and practice reality regarding the carrying out of consumerist principles are incongruous. Interactions contained relatively simple information, were dominated and led by nurses and offered little opportunity for patient choice. Patients, however, expressed a satisfaction with minimal information and involvement. CONCLUSION: The findings are discussed with reference to a number of different contextual factors: acuity of illness, perceived balance of power, information gaps, patterns of contact and nurse-patient relationships, and patient-centred care. PMID- 11472513 TI - Conceptions of life situation among next-of-kin of haemodialysis patients. AB - The presence of renal disease in a family is a strain on both the patient and the next-of-kin, affecting their life situations. Surprisingly, few studies dealing with the ways that the next-of-kin experience their situation are available. The aim of this study was to describe how the next-of-kin of haemodialysis patients conceive their life situation. Data were collected by interviewing 12 people who live with someone with dialysis-treated renal disease and analysed according to a qualitative method inspired by the phenomenographic approach. Six description categories of how the subjects construed their life situation emerged: a feeling of confinement; a feeling of social isolation; a feeling that the way of life has changed; a feeling of security in life; a feeling of a threatening future; and promoting health. The next-of-kin generally expressed a large degree of commitment to and concern for the sick person. In spite of their life situation having been dramatically changed, the next-of-kin described an ability to adapt. With the help of society the feelings of confinement and social isolation can be dispersed, enabling the next-of-kin to promote the health of the sick person. A suggestion for further research is to study what adaptation strategies next-of kin use in their life situation. PMID- 11472514 TI - Growth of Escherichia coli O157:H7 during sprouting of alfalfa seeds. AB - AIMS: Escherichia coli O157:H7 was monitored daily during sprouting of alfalfa seeds inoculated at high (3.92 log10 cfu g(-1)) and low (1.86 log10 cfu g(-1)) levels to assess the extent of pathogen growth during production. METHODS AND RESULTS: Sprouts and rinse water were tested by direct and membrane filter plating on modified sorbitol MacConkey agar and BCM O157:H7(+) agar; the antibody direct epifluorescent filter technique; and rapid immunoassays. The pathogen reached maximum populations after one and two days of sprouting seeds inoculated at high and low levels, respectively; in either case, populations of 5-6 log10 cfu g(-1) were reached. Detection limits of two rapid immunoassays, Reveal and VIP, without enrichment were determined to be 5-7 log10 cfu ml(-1). CONCLUSION: These results show the ability of E. coli O157:H7 to grow to high levels during sprouting; however, because these levels may be below detection limits, it is necessary to include enrichment when monitoring sprout production for E. coli O157:H7 by the rapid test kits. SIGNIFICANCE AND IMPACT OF THE STUDY: The data indicate that sprouts may harbor high levels of pathogens. The appropriate use of rapid test methods for pathogen monitoring during sprouting is indicated. PMID- 11472515 TI - Evaluation of spore extraction and purification methods for selective recovery of viable Bacillus anthracis spores. AB - AIMS: To investigate methods of improving anthrax spore detection with PLET. METHODS AND RESULTS: Comparisons were made of PLET and blood-supplemented PLET to recover and distinguish spores of a variety of Bacillus species. Heat and ethanol purification of spores, and spore extraction from soil with water and high specific gravity sucrose plus non-ionic detergent, were also carried out. CONCLUSION: PLET was more selective and suitable than blood-supplemented PLET for detection of anthrax spores in the environmental specimens. However, PLET is not an optimal spore recovery medium. Purification of spores with ethanol was as effective as heat purification. High specific gravity sucrose plus detergent extraction solutions may be more sensitive than extraction with water. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights shortcomings with the standard PLET isolation of anthrax spores and describes ways in which the procedure may be improved. PMID- 11472516 TI - Genetic diversity of Saccharomyces cerevisiae strains during the 24 h fermentative cycle for the production of the artisanal Brazilian cachaca. AB - AIMS: Characterization of yeast populations and genetic polymorphism of Saccharomyces cerevisiae strains collected during the short fermentative cycles from the spontaneous fermentations during the artisanal cachaca production. METHODS AND RESULTS: The prevalent S. cerevisiae strains were analysed by PFG and RAPD-PCR using primers EI1 and M13. The molecular analysis have showed a high degree of genetic polymorphism among the strains within a 24 h fermentative cycle. CONCLUSION: The genetic diversity observed in the S. cerevisiae strains may be occurring due to the existence of a large number of individual genotypes within the species. The unique characteristics of the cachaca fermentation process probably allows for a faster detection of molecular polymorphisms of yeast strains than other types of fermentations. SIGNIFICANCE AND IMPACT OF THE STUDY: Spontaneous fermentations to produce cachaca, due to their characteristics, are an excellent model for the study of molecular diversity of S. cerevisiae strains during the production of fermented beverages. PMID- 11472517 TI - Breeding of mycoparasitic Trichoderma strains for heavy metal resistance. AB - AIMS: This study was designed to investigate the effects of 10 heavy metals on the in vitro activities of beta-glucosidase, cellobiohydrolase, beta-xylosidase and endoxylanase enzymes for six strains of Trichoderma, and to isolate and characterize heavy metal-resistant mutants. METHODS AND RESULTS: At a concentration of 1 mmol, only mercury showed significant inhibitory effects on the in vitro enzyme activities; in all other cases, the enzymes remained active. A total of 177 heavy metal-resistant mutants were isolated and tested for cross resistance to other heavy metals. Some mutants were effective antagonists of Fusarium, Pythium and Rhizoctonia strains, even on media containing the respective heavy metals. CONCLUSION: Trichoderma strains could be developed as biocontrol agents that are effective against plant pathogenic fungi, even under heavy metal stress. SIGNIFICANCE AND IMPACT OF THE STUDY: Trichoderma mutants resistant to heavy metals might be of value for use with heavy metal-containing pesticides, as part of an integrated plant protection system. PMID- 11472518 TI - Characterization of pectin lyase produced by an endophytic strain isolated from coffee cherries. AB - AIMS: The effect of endophytic bacterial activity on the quality of coffee beverage was studied. METHODS AND RESULTS: A survey of the micro-organisms in coffee cherries was performed before harvesting, and their growth on the main nutrients available in coffee cherries was determined in vitro. CONCLUSION: Many endophytic bacteria were isolated from surface-sterilized coffee cherries. One of the pectinolytic strains was physiologically and phenotypically characterized, and was tentatively identified by partial 16S rDNA sequencing as Paenibacillus amylolyticus. This endophytic strain produced an extracellular pectinase with maximal activity at 40 degrees C and pH 7.9, and was thermostable up to 45 degrees C. EDTA and metal ions had little effect on pectin lyase activity. Km and Vmax values were 4.6 mg ml(-1) and 94.0 10(-8) mol min(-1) ml(-1), respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: Pectin lyases have been found in fungi but rarely in bacteria, and this isolate is a promising tool for regulation studies of these enzymes. PMID- 11472519 TI - Evaluation of co-aggregation among Streptococcus mitis, Fusobacterium nucleatum and Porphyromonas gingivalis. AB - AIMS: To develop a semi-quantitative method for evaluating co-aggregation reactions among three bacterial species, and to examine the influence of Fusobacterium nucleatum on the adherence of Porphyromonas gingivalis. METHODS AND RESULTS: The method involves coating hydroxyapatite (HAP) discs with streptococcal cells and treatment with radio-labelled bacterial cell suspensions. The sensitivity of the method was estimated by comparison with a turbidometric co aggregation assay. Results from the two methods were in close agreement. Streptococcus mitis-coated HAP discs were immersed in a 3H-labelled Fus. nucleatum cell suspension and then a 14C-labelled P. gingivalis cell suspension. The discs were then pyrolysed to recover and quantify the released 3H and 14C radioactivity. The number of Fus. nucleatum cells on the discs increased with immersion time and this, in turn, resulted in elevated adherence of P. gingivalis. CONCLUSION: The data indicate that the method closely reflects co aggregation characters, and that Fus. nucleatum has a positive effect on the adherence of P. gingivalis. SIGNIFICANCE AND IMPACT OF THE STUDY: The present method, which is designed to mimic the oral environment, should prove useful in the semi-quantitative evaluation of co-aggregation reactions. PMID- 11472520 TI - Improved acid tolerance of a recombinant strain of Escherichia coli expressing genes from the acidophilic bacterium Oenococcus oeni. AB - AIMS: Oenococcus oeni is a lactic acid bacterium used in wine fermentation. Two open reading frames (orfB and orfC) were identified in the upstream region of the hsp18 gene, encoding the small heat-shock protein Lo18. Expression of these genes in conditions of acid stress was studied in Escherichia coli. METHODS AND RESULTS: Sequence analysis showed that orfB encodes a putative transcriptional regulator of the LysR family. The protein encoded by orfC shares homologies with multi-drug resistance systems. Heterologous expression of orfB, orfC and hsp18 genes in Escherichia coli significantly enhanced the viability of the host strain under acidic conditions. CONCLUSION: It was demonstrated that the three genes were needed for acquisition of this acid tolerance phenotype. SIGNIFICANCE AND IMPACT OF THE STUDY: Heterologous expression of Oenococcus genes could be used to confer acidophilic behaviour on strains of biotechnological interest. PMID- 11472521 TI - Survival of faecal indicator bacteria in bovine manure incorporated into soil. AB - AIMS: Survival of Escherichia coli and enterococci was evaluated in bovine manure incorporated into two Wisconsin soils. METHODS AND RESULTS: Silty clay loam (SCL) and loamy sand (LS) were mixed with fresh bovine manure, exposed daily to 10 h at 22 degrees C/14 h at 9 degrees C, and watered weekly for 12 weeks. Escherichia coli numbers increased 1-2 log cfu g(-1), then decreased < 1 and about 2 log cfu g(-1) in SCL and LS, respectively. Enterococci numbers rose less and then declined faster than those of E. coli. Watering intervals of 3, 7 and 14 days were evaluated in weeks 13-19, but did not affect the slow decline in numbers of E. coli or enterococci. CONCLUSION: Escherichia coli and enterococci may survive at least 19 weeks at 9-21 degrees C in bovine manure/soil, with E. coli surviving better. SIGNIFICANCE AND IMPACT OF THE STUDY: Quantification of E. coli or enterococci in late spring/early summer soil may be useful in indicating recent application of bovine manure. PMID- 11472522 TI - Diversity of Vibrio spp. populations in several exhibition aquaria with a shared water supply. AB - AIMS: Abiotic factors may influence the settlement of bacterial populations in similar marine environments. Exhibition aquaria are a model for the study of the settlement of bacteria in different environment. Vibrio populations in the seawater reservoir, the Mediterranean tank and the Tropical tank from an exhibition aquarium on the western coast of the Mediterranean were compared and the effect of abiotic factors on the structure of these populations was considered. METHODS AND RESULTS: High diversity indexes and similar Vibrio populations were found in the water of the reservoir and of the Mediterranean tank, whereas a lower diversity and different main populations were found in the water of the Tropical tank. The antibiotic resistance profiles of the most representative strains, presented a number of differences depending on the origin of the sample. CONCLUSION: Abiotic conditions, mainly temperature, may determine the structure and composition of Vibrio populations in exhibition aquaria. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial monitoring of water could be useful for health management of aquatic environments. PMID- 11472523 TI - Mycotoxins inactivation by extrusion cooking of corn flour. AB - AIMS: To evaluate the effects of the extrusion cooking process on the inactivation of mycotoxins in corn flour. METHODS AND RESULTS: Samples of corn flour experimentally contaminated with aflatoxin B1 (AFB1) (50 ppb) and deoxynivalenol (DON) (5 ppm) were extruded. The effects of three extrusion variables (flour moisture, extrusion temperature and sodium metabisulphite addition) were analysed according to a two-level factorial design. The process was effective for the reduction of DON content (higher than 95%) under all the conditions assessed, but was only partially successful (10-25%) for the decontamination of AFB1. CONCLUSION: Extrusion cooking is effective for the inactivation of DON but is of limited value for AFB1, even if metabisulphite is added. More severe extrusion conditions are needed for the detoxification of AFB1. SIGNIFICANCE AND IMPACT OF THE STUDY: As contamination with DON occurs mainly in the field prior to harvesting and that of AFB1 is normally produced during grain storage, maize is often contaminated with DON but not with AFB1. Under these conditions, the described extrusion process can be used for the detoxification of DON. The addition of sodium metabisulphite did not significantly affect the inactivation of AFB1. Extrusion cooking is therefore an appropriate treatment for vomitoxin-contaminated maize in countries where, because of the prevailing conditions, these are the only toxins present. PMID- 11472524 TI - Influence of copper-alloying of austenitic stainless steel on multi-species biofilm development. AB - AIMS: To investigate the bactericidal influence of copper-alloying of stainless steel on microbial colonization. METHODS AND RESULTS: Inhibition of bacterial adherence was investigated by monitoring (192 h) the development of a multi species biofilm on Cu-alloyed (3.72 wt%) stainless steel in a natural surface water. During the first 120 h of exposure, lower numbers of viable bacteria in the water in contact with copper-containing steel relative to ordinary stainless steel were observed. Moreover, during the first 48 h of exposure, lower colony counts were found in the biofilm adhering to the Cu-alloyed steel. No lower colony or viable counts were found throughout the remainder of the experimental period. CONCLUSION: The presence of Cu in the steel matrix impedes the adhesion of micro-organisms during an initial period (48 h), while this bactericidal effect disappears after longer incubation periods. SIGNIFICANCE AND IMPACT OF THE STUDY: The application of Cu-alloyed stainless steels for bactericidal purposes should be restricted to regularly-cleaned surfaces. PMID- 11472525 TI - Partial characterization of a bacteriocin produced by Lactobacillus helveticus. AB - AIMS: To investigate the antimicrobial activity of a strain of Lactobacillus helveticus. METHODS AND RESULTS: The culture supernatant fluid Lact. helveticus G51 showed antimicrobial activity against thermophilic strains of Lactobacillus. Purification of the active compound was achieved after gel filtration and ion exchange chromatography. As revealed by SDS-PAGE, active fractions were relatively homogeneous, showing a protein with a molecular mass of 12.5 kDa. The antimicrobial compound was heat labile, inactivated by proteolytic enzymes and had a bactericidal mode of action. CONCLUSION: The antimicrobial activity expressed by Lact. helveticus G51 was correlated with the production of a bacteriocin with properties that were different to other helveticins. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has provided further data on Lact. helveticus bacteriocins. The strong activity of the bacteriocin towards various thermophilic lactobacilli warrants further investigation for its potential to obtain attenuated cultures for the enhancement of the cheese ripening process. PMID- 11472526 TI - Fermentation of pectin and glucose, and activity of pectin-degrading enzymes in the rumen bacterium Lachnospira multiparus. AB - AIMS: Lachnospira multiparus belongs to the main rumen pectinolytic bacteria. Its carbohydrate metabolism was studied in growth experiments on laboratory fermenters, and using assays of activities of enzymes involved in pectin fermentation. METHODS AND RESULTS: The type strain of this species and two substrates were used. Lachnospira multiparus ATCC 19207 grew on pectin and glucose at a similar rate and had no preference for one or the other substrate. Pectin-grown cultures, however, produced significantly more acetate and less formate, lactate, ethanol, hydrogen, cell dry matter and protein than corresponding cultures grown on glucose. Extracellular exopectate lyase (EC 4.2.2.9) was the principal enzyme degrading the pectin macromolecule. Cell extracts possessed 2-keto-3- deoxy-6-phosphogluconate aldolase (EC 4.1.2.14) and fructosediphosphate aldolase (EC 4.1.2.13) activity. The former enzyme catalyses the final reaction in the Entner-Doudoroff pathway; the latter is the key enzyme of glycolysis and the pentose phosphate pathway. CONCLUSION: These results are consistent with the assumption that acidic products of pectin degradation are catabolized via a modified Entner-Doudoroff pathway. Phosphogluconate was not metabolized by cell extracts of the strain studied. SIGNIFICANCE AND IMPACT OF THE STUDY: This suggests that the conventional Entner-Doudoroff pathway of glucose utilization does not operate in this bacterium, presumably because of the lack of 6-phosphogluconate dehydrase (EC 4.2.1.12) activity. PMID- 11472527 TI - Effect of copper on the degradation of phenanthrene by soil micro-organisms. AB - AIMS: The effect of copper on the degradation by soil micro-organisms of phenanthrene, a polycyclic aromatic hydrocarbon, was investigated. METHODS AND RESULTS: Inert nylon filters were incubated in the soil for 28 days at 25 degrees C. Each filter was inoculated with a soil suspension, phenanthrene (400 ppm), copper (0, 70, 700 or 7000 ppm) and nitrogen/phosphorus sources. The filters were assessed for phenanthrene degradation, microbial respiration and colonization. Phenanthrene degradation proceeded even at toxic copper levels (700/7000 ppm), indicating the presence of phenanthrene-degrading, copper-resistant and/or tolerant microbes. However, copper at these high levels reduced microbial activity (CO2 evolution). CONCLUSION: High levels of copper caused an incomplete mineralization of phenanthrene and possible accumulation of its metabolites. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of heavy metals in soils could seriously affect the bioremediation of PAH-polluted environments. PMID- 11472528 TI - Polysaccharide production by a reduced pigmentation mutant of Aureobasidium pullulans NYS-1. AB - AIMS: To isolate a reduced pigmentation mutant of Aureobasidium pullulans NYS-1 and characterize its cellular pigmentation plus its polysaccharide and biomass production relative to carbon source. METHODS AND RESULTS: Cellular pigmentation, polysaccharide levels and biomass production by the isolated mutant NYSRP-1 were analysed relative to carbon source. Cellular pigmentation of the mutant was lower than its parent strain using either carbon source. The mutant elaborated higher polysaccharide levels on sucrose than on corn syrup. The pullulan content of the polysaccharide synthesized and biomass production by the mutant rose as the carbon source concentration was increased. CONCLUSION: It is feasible to isolate a reduced pigmentation mutant from strain NYS-1 that exhibits elevated polysaccharide production using corn syrup as a carbon source. SIGNIFICANCE AND IMPACT OF THE STUDY: The mutant provides an advantage for commercial pullulan production because of its reduced pigmentation and enhanced polysaccharide synthesis. PMID- 11472529 TI - Haploid chromosomes in molecular ecology: lessons from the human Y. AB - We review the potential use of haploid chromosomes in molecular ecology, using recent work on the human Y chromosome as a paradigm. Chromosomal sex determination systems, and hence constitutively haploid chromosomes, which escape from recombination over much of their length, have evolved multiple times in the animal kingdom. In mammals, where males are the heterogametic sex, the patrilineal Y chromosome represents a paternal counterpart to mitochondrial DNA. Work on the human Y chromosome has shown it to contain the same range of polymorphic markers as the rest of the nuclear genome and these have rendered it the most informative haplotypic system in the human genome. Examples from research on the human Y chromosome are used to illustrate the common interests of anthropologists and ecologists in investigating the genetic impact of sex specific behaviours and dispersals, as well as patterns of global diversity. We present some methodologies for extracting information from these uniquely informative yet under-utilized loci. PMID- 11472530 TI - Assessment of genetic structure within and among Bulgarian populations of the common ash (Fraxinus excelsior L.). AB - We analysed genetic variation within and between populations of the common ash from Bulgaria in order to extract biological information useful in the context of conservation management of eastern European genetic resources of noble hardwood species. A total of 321 trees from three regions of Bulgaria were typed at six highly polymorphic microsatellite loci. Analysis of within-population inbreeding suggests an upper boundary value of 2.7% for the selfing rate. Significant spatial genetic structure consistent with models of isolation by distance was detected within four out of 10 populations as well as among populations. Estimates of neighbourhood size in the range 38-126 individual trees were obtained based on spatial genetic structure analyses at either the intrapopulation or interpopulation level. Differentiation among populations explained only about 8.7% of total genetic diversity. These results are discussed in comparison with data from social broad-leaved species such as oak and beech. PMID- 11472531 TI - Microgeographic genetic structure in the yellow-pine chipmunk (Tamias amoenus). AB - While there is evidence for broad-scale genetic structure in small mammals, few studies have used variable DNA-based genetic markers to examine genetic differentiation at microgeographic (tens of kilometres) scales. Yellow-pine chipmunks (Tamias amoenus) live in the heterogeneous landscape of the Rockies in southwest Alberta and are generally restricted to areas of low elevation. We used seven microsatellite loci to determine whether chipmunks show evidence of population genetic structure among three closely situated sites (< 15 km) in the Kananaskis Valley, Alberta. We found evidence for genetic structure in the form of significant differences in allele frequencies among populations and significantly nonzero values of FST for both overall and pairwise population comparisons. However, FIS values for each population were not significantly different from zero, suggesting little evidence for inbreeding within populations. Genetic differentiation probably occurs as a result of the strong effect of drift in very small (N(e) approximately 25) populations of these animals even in the face of substantial immigration rates. PMID- 11472532 TI - Microsatellite variation in the yellowhammer Emberiza citrinella: population structure of a declining farmland bird. AB - In recent years, there has been much concern in the UK about population declines of widespread species in agricultural habitats. Conservation-orientated research on declining birds has focused on vital rates of survival and productivity. However, the environmental factors which may influence movements between populations of widespread species is poorly understood. Population genetic structure is an indirect description of dispersal between groups of individuals. To attempt to develop an understanding of genetic structuring in a widespread, but declining, farmland bird, we therefore investigated the yellowhammer, Emberiza citrinella, population in England and Wales using microsatellite data. Our first aim was to investigate whether there was genetic substructuring in the population. A second aim was to investigate if there was a relationship between genetic distances and various environmental variables. Finally, we analysed the microsatellite data for evidence of loss of genetic variation due to population decline. Our data showed a slight but significant structure within the yellowhammer population. This therefore cannot be considered a panmictic population. Our example from South Cumbria implies that high-altitude barriers may have a slight influence on population structure. However, on the whole, genetic distances between sample sites were not significantly correlated with geographical distances, degrees of population connectivity, high altitudes, or differences in precipitation between sites. Finally, we detected departures from mutation-drift equilibrium (excess heterozygosity), which is indicative of a loss of genetic variation through recent decline. PMID- 11472533 TI - Gene flow on the ice: genetic differentiation among Adelie penguin colonies around Antarctica. AB - Each summer Adelie penguins breed in large disjunct colonies on ice-free areas around the Antarctic continent. Comprising > 10 million birds, this species represents a dominant feature of the Antarctic ecosystem. The patchy distribution within a large geographical range, natal philopatry and a probable history of refugia, suggest that this species is likely to exhibit significant genetic differentiation within and among colonies. We present data from seven microsatellite DNA loci for 442 individuals from 13 locations around the Antarctic continent. With the exception of one locus, there was no significant genic or genotypic heterogeneity across populations. Pairwise FST values were low with no value > 0.02. When all colonies were compared in a single analysis, the overall FST value was 0.0007. Moreover, assignment tests were relatively ineffective at correctly placing individuals into their respective collection sites. These data reveal a lack of genetic differentiation between Adelie penguin colonies around the Antarctic continent, despite substantial levels of genetic variation. We consider this homogeneity in terms of the dispersal of individuals among colonies and the size of breeding groups and discuss our results in terms of the glacial history of Antarctica. PMID- 11472534 TI - Population structure in the endangered Mauna Loa silversword, Argyroxiphium kauense (Asteraceae), and its bearing on reintroduction. AB - Reintroduction of populations of endangered species is a challenging task, involving a number of environmental, demographic and genetic factors. Genetic parameters of interest include historical patterns of genetic structure and gene flow. Care must be taken during reintroduction to balance the contrasting risks of inbreeding and outbreeding depression. The Mauna Loa silversword, Argyroxiphium kauense, has experienced a severe decline in population size and distribution in the recent past. Currently, three populations with a total of fewer than 1000 individuals remain. We measured genetic variation within and among the remnant populations using seven microsatellite loci. We found significant genetic variation remaining within all populations, probably related to the recent nature of the population impact, the longevity of the plants, and their apparent self-incompatibility. We also found significant genetic differentiation among the populations, reinforcing previous observations of ecological and morphological differentiation. With respect to reintroduction, the results suggest that, in the absence of additional data to the contrary, inbreeding depression may not be a substantial risk as long as propagules for the founding of new populations are adequately sampled from within each source population before additional inbreeding takes place. The results further suggest that if mixing of propagules from different source populations is not required to increase within-population genetic variation in the reintroduced populations, it may best be avoided. PMID- 11472535 TI - Genetic differentiation in Eurasian populations of the postfire ascomycete Daldinia loculata. AB - The genetic population structure of the postfire ascomycete Daldinia loculata was studied to test for differentiation on a continental scale. Ninety-six samples of spore families, each comprising mycelia from six to 10 spores originating from single perithecia, were sampled from one Russian and six Fennoscandian forest sites. Allelic distribution was assayed for six nuclear gene loci by restriction enzyme analyses of polymerase chain reaction (PCR)-amplified gene fragments. In addition, the full sequence of the gene fragment was analysed for a subset of haploid single-ascospore isolates in a multiallelic approach. A third data set was generated by using arbitrary-primed PCR with the core sequence of the phage M13 as primer. Although there was a reduction in heterozygosity in the total population from what would have been expected at random mating, the levels of genetic differentiation among the Eurasian subpopulations of D. loculata were low. All subpopulations were found to be in Hardy-Weinberg equilibrium and gametic equilibrium was observed between all investigated nuclear gene loci. The results obtained by the different markers were consistent; we confirmed low levels of genetic differentiation among the Eurasian subpopulations of D. loculata. The differentiation did not increase with distance; the Russian subpopulation, sampled more than 7000 km from the Fennoscandian subpopulations, was only moderately differentiated from the others (FST = 0.00-0.14). In contrast, one of the Swedish populations was the most highly differentiated from the others, with FST and GST values of 0.10-0.16. The results suggest that D. loculata consists of a long-lived background Eurasian population of latent mycelia in nonburned forests, established by sexual ascospores dispersed from scattered burned forest sites. Local differentiation is probably due to founder effects of populations in areas with low fire frequency. A tentative life cycle of D. loculata is presented. PMID- 11472536 TI - Characterization of major histocompatibility complex DRB diversity in the endemic South African antelope Damaliscus pygargus: a comparison in two subspecies with different demographic histories. AB - Major histocompatibility complex (MHC) class II locus DRB was investigated by single-strand conformation polymorphism analysis (SSCP) and sequence analysis in the endemic South African antelope, Damaliscus pygargus, of which there are two subspecies. Greater polymorphism was found in the blesbok (D. p. phillipsi) subspecies (n = 44; 22 alleles) than in the bontebok (D. p. pygargus) subspecies (n = 45; 6 alleles). Erosion of allelic diversity in bontebok was most likely the result of two severe bottleneck events caused by hunting pressure and parasitic infection. A majority of the polymorphism observed was found within the peptide binding region (PBR) where dN/dS ratios were higher than for the non-PBR region. This, and the apparent trans-species relationship among alleles in a bovid phylogeny, suggest the evolution of diversity by heterosis or frequency-dependent selection. PMID- 11472537 TI - Molecular investigations in populations of Spartina anglica C.E. Hubbard (Poaceae) invading coastal Brittany (France). AB - Spartina anglica is a classical example of recent alloploid speciation. It arose during the end of the nineteenth century in England by hybridization between the indigenous Spartina maritima and the introduced East-American Spartina alterniflora. Duplication of the hybrid genome (Spartina x townsendii) gave rise to a vigorous allopolyploid involved in natural and artificial invasions on different continents. Spartina anglica was first recorded in France in 1906, and since then, it has spread all along the western French coast. Earlier studies revealed that native British populations display consistent morphological plasticity and lack of isozyme variation. In this paper, we use different molecular markers (randomly amplified polymorphic DNA, intersimple sequence repeats and restriction patterns from nuclear and chloroplast DNA sequences) to analyse the genetic patterns of the French populations of S. anglica. Our results show that French populations are mainly composed of one "major" multilocus genotype. This genotype is identical to the first-generation hybrid S. x townsendii from England. Losses of few markers from this genotype are observed but are restricted to a few populations from Brittany; it is likely that they appeared independently, subsequent to their introduction. In southern Brittany, no hybrids between S. anglica and S. maritima have been found where the two species co-occur. All French populations of S. anglica display the same chloroplast DNA sequences as S. alterniflora, the maternal genome donor. These findings are consistent with a severe genetic bottleneck at the time of the species formation, as a consequence of a unique origin of the species. Both parental nuclear sequences are present in the allopolyploid populations, revealing that for the markers investigated, no extensive changes have occurred in this young species. PMID- 11472538 TI - Patterns of population subdivision, gene flow and genetic variability in the African wild dog (Lycaon pictus). AB - African wild dogs are large, highly mobile carnivores that are known to disperse over considerable distances and are rare throughout much of their geographical range. Consequently, genetic variation within and differentiation between geographically separated populations is predicted to be minimal. We determined the genetic diversity of mitochondrial DNA (mtDNA) control region sequences and microsatellite loci in seven populations of African wild dogs. Analysis of mtDNA nucleotide diversity suggests that, historically, wild dog populations have been small relative to other large carnivores. However, population declines due to recent habitat loss have not caused a dramatic reduction in genetic diversity. We found one historical and eight recent mtDNA genotypes in 280 individuals that defined two highly divergent clades. In contrast to a previous, more limited, mtDNA analysis, sequences from these clades are not geographically restricted to eastern or southern African populations. Rather, we found a large admixture zone spanning populations from Botswana, Zimbabwe and south-eastern Tanzania. Mitochondrial and microsatellite differentiation between populations was significant and unique mtDNA genotypes and alleles characterized the populations. However, gene flow estimates (Nm) based on microsatellite data were generally greater than one migrant per generation. In contrast, gene flow estimates based on the mtDNA control region were lower than expected given differences in the mode of inheritance of mitochondrial and nuclear markers which suggests a male bias in long-distance dispersal. PMID- 11472539 TI - DNA analysis of transferred sperm reveals significant levels of gene flow between molecular forms of Anopheles gambiae. AB - Anopheles gambiae populations in west Africa are complex, being composed of multiple, sympatric subpopulations. Recent studies have failed to reveal significant genetic differences among subpopulations, stimulating a debate regarding the levels of gene flow among them. The observed homogeneity may be the consequence of substantial contemporary gene flow or it may be that reproductive isolation is complete, but too recent for the accumulation of significant levels of genic divergence. Here, we report the results of a study estimating contemporary levels of gene flow between An. gambiae subpopulations by analysing females and transferred sperm removed from their reproductive systems. A total of 251 female and associated sperm extracts was analysed from a single site in Mali. Two molecular forms of An. gambiae, the M- and S-forms, occurred in sympatry at this site. Overall, we found very strong positive assortative mating within forms, however, we did observe significant hybridization between forms. In the M subpopulation 2/195 females (1.03%) contained sperm from S-form males and in 55 S form females we found one female containing M-form sperm (1.82%). We also identified a mated M xS hybrid adult female. From mating frequencies, we estimate the Nem between the M- and S-form at 16.8, and from the adult hybrid frequency at 5.6. These values are consistent with our earlier estimate, based on FST for 21 microsatellite loci in which Nem = 5.8. We conclude that the general lack of genetic divergence between the M and S subpopulations of An. gambiae can be explained entirely by contemporary gene flow. PMID- 11472540 TI - Genetic structure and distinctness of Apis mellifera L. populations from the Canary Islands. AB - The genetic structure of Apis mellifera populations from the Canary Islands has been assessed by mitochondrial (restriction fragment length polymorphisms of the intergenic transfer RNAleu-COII region) and nuclear (microsatellites) studies. These populations show a low level of genetic variation in terms of average number of alleles and degree of heterozygosity. Significant differences in the distribution of alleles were found in both data sets, confirming the genetic differentiation among some of the islands but not within them. Two mitochondrial haplotypes characteristic of the Canary Islands are found at high frequencies, although populations are introgressed by imported honeybees of eastern European C lineage. This introgression is rather high on Tenerife and El Hierro and low on Gran Canaria and La Gomera, whereas on La Palma it has not been recorded. The finding of microsatellite alleles characteristic of the eastern European lineage corroborates the genetic introgression. Phylogenetic analyses indicate that the Canarian honeybees are differentiated from other lineages and provide genetic evidence of their African origin. PMID- 11472541 TI - Phylogeography of the intertidal copepod Tigriopus californicus reveals substantially reduced population differentiation at northern latitudes. AB - Previous studies of the intertidal copepod Tigriopus californicus revealed one of the highest levels of mitochondrial DNA differentiation ever reported among conspecific populations. The present study extends the geographical sampling northward, adding populations from northern California to south-east Alaska. The mitochondrial phylogeny for the entire species range, based on cytochrome oxidase I sequences for a total of 49 individuals from 27 populations, again shows extreme differentiation among populations (up to 23%). However, populations from Oregon northwards appear to be derived and have interpopulation divergences five times lower than those between southern populations. Furthermore, although few individuals were sequenced from each locality, populations from Puget Sound northward had significantly reduced levels of within-population variation. These patterns are hypothesized to result from the contraction and expansion of populations driven by recent ice ages. PMID- 11472542 TI - Phylogeography of the Vosges mountains populations of Gonioctena pallida (Coleoptera: Chrysomelidae): a nested clade analysis of mitochondrial DNA haplotypes. AB - The pattern of genetic variation in the leaf beetle Gonioctena pallida was investigated inside the Vosges mountains using a highly variable 363 bp DNA fragment of the mitochondrial control region. Sequencing of 242 individuals, sampled in a geographical area of 100 x 40 km, identified 61 haplotypes whose genealogy was inferred. The resulting haplotype network exhibits four star-like phylogenies, two of which may be indicative of a population having recently expanded in size from a small number of founders. Nested clade analysis suggested multiple past expansion events, but also isolation by distance and possibly past fragmentation events, as the causes of the detected geographical associations of haplotypes. These results indicate the existence of effective barriers to gene flow inside the investigated area. Because the oldest demographic events inferred in the nested clade analysis were identified as expansion events, we hypothesize that a small population of founders have expanded not only in size, but also in geographical range from the south towards the north and east of the Vosges. PMID- 11472543 TI - Species boundaries and genetic diversity among Hawaiian crickets of the genus Laupala identified using amplified fragment length polymorphism. AB - Crickets of the genus Laupala represent one of the many morphologically cryptic groups of insects, with the most closely related species distinguished only by the male calling song. Cryptic groups provide a challenge in determining the genetic boundaries between closely related populations and species. We have addressed the question of species boundaries in the Hawaiian cricket, Laupala, using nuclear DNA patterns sampled by the amplified fragment length polymorphism (AFLP) technique. This method has been used widely by plant researchers to facilitate the rapid assessment of genetic diversity in very closely related species and varieties. The AFLP technique is simple and robust, can be applied to any organism, and overcomes problems associated with cost, development time, information content and reproducibility that can plague other marker systems. Our results support previously hypothesized taxonomic relationships among sympatric populations and suggest close genetic relationships among allopatric, conspecific populations. PMID- 11472544 TI - Molecular differentiation of the Old World Culicoides imicola species complex (Diptera, Ceratopogonidae), inferred using random amplified polymorphic DNA markers. AB - Samples of seven of the 10 morphological species of midges of the Culicoides imicola complex were considered. The importance of this species complex is connected to its vectorial capacity for African horse sickness virus (AHSV) and bluetongue virus (BTV). Consequently, the risk of transmission may vary dramatically, depending upon the particular cryptic species present in a given area. The species complex is confined to the Old World and our samples were collected in Southern Africa, Madagascar and the Ivory Coast. Genomic DNA of 350 randomly sampled individual midges from 19 populations was amplified using four 20-mer primers by the random amplified polymorphic DNA (RAPD) technique. One hundred and ninety-six interpretable polymorphic bands were obtained. Species specific RAPD profiles were defined and for five species diagnostic RAPD fragments were identified. A high degree of polymorphism was detected in the species complex, most of which was observed within populations (from 64 to 76%). Principal coordinate analysis (PCO) and cluster analysis provided an estimate of the degree of variation between and within populations and species. There was substantial concordance between the taxonomies derived from morphological and molecular data. The amount and the different distributions of genetic (RAPD) variation among the taxa can be associated to their life histories, i.e. the abundance and distribution of the larval breeding sites and their seasonality. PMID- 11472545 TI - Ecology and population genetics of Sonoran Desert Drosophila. AB - Three species of cactophilic Drosophila endemic to the Sonoran Desert of North America, D. nigrospiracula, D. pachea and D. mettleri, experience marked differences in spatial resource availability, and the first two of these display significant differences in dispersal behaviour. We employed starch gel and cellulose acetate electrophoresis for eight allozyme loci to test for a relationship between these variables and genetic differentiation among geographical populations of each species. No evidence was found for population structure in any of the three species, populations of which were separated by geographical distances of up to 475 km. Allele frequencies for two loci, Mdh-1 and Est-2, in D. nigrospiracula and D. pachea were very similar to those obtained approximately 30 years ago by other workers, indicating that the polymorphisms are remarkably stable under the stressful and variable conditions of the desert environment. High longevity, dispersal and multiple female remating are likely to contribute to the apparent high level of gene flow in all three species. PMID- 11472546 TI - The genetic mating system and tests for cuckoldry in a pipefish species in which males fertilize eggs and brood offspring externally. AB - Highly variable microsatellite loci were used to study the mating system of Nerophis ophidion, a species of pipefish in which pregnant males carry embryos on the outside of their body rather than in an enclosed brood pouch. Despite this mode of external fertilization and brooding, otherwise rare in the family Syngnathidae, the genotypes of all embryos proved to be consistent with paternity by the tending male, thus indicating that cuckoldry by sneaker males is rare or nonexistent in this species. N. ophidion is a phylogenetic outlier within the Syngnathidae and its reproductive morphology is thought to be close to the presumed ancestral condition for pipefishes and seahorses. Thus, our genetic results suggest that the evolutionary elaboration of the enclosed brood pouch elsewhere in the family was probably not in response to selection pressures on pregnant males to avoid fertilization thievery. With regard to maternity assignments, our genotypic data are consistent with behavioural observations indicating that females sometimes mate with more than one male during a breeding episode, and that each male carries eggs from a single female. Thus, the polyandrous genetic mating system in this species parallels the social mating system, and both are consistent with a more intense sexual selection operating on females, and the elaboration of secondary sexual characters in that gender. PMID- 11472547 TI - Diversity of endophytic fungi of single Norway spruce needles and their role as pioneer decomposers. AB - The diversity of endophytic fungi within single symptomless Norway spruce needles is described and their possible role as pioneer decomposers after needle detachment is investigated. The majority (90%) of all 182 isolates from green intact needles were identified as Lophodermium piceae. Up to 34 isolates were obtained from single needles. Generally, all isolates within single needles had distinct randomly amplified microsatellite (RAMS) patterns. Single trees may thus contain a higher number of L. piceae individuals than the number of their needles. To investigate the ability of needle endophytes to act as pioneer decomposers, surface-sterilized needles were incubated on sterile sand inoculated with autoclaved or live spruce forest humus layer. The dry weight loss of 13-17% found in needles after a 20-week incubation did not significantly differ between the sterilized and live treatments. Hence, fungi surviving the surface sterilization of needles can act as pioneer decomposers. A considerable portion of the needles remained green during the incubation. Brown and black needles, in which the weight loss had presumably taken place, were invaded throughout by single haplotypes different from L. piceae. Instead, Tiarasporella parca, a less common needle endophyte, occurred among these invaders of brown needles. Needle endophytes of Norway spruce seem thus to have different abilities to decompose host tissues after needle cast. L. piceae is obviously not an important pioneer decomposer of Norway spruce needles. The diversity of fungal individuals drops sharply when needles start to decompose. Thus, in single needles the decomposing mycota is considerably less diverse than the endophytic mycota. PMID- 11472548 TI - Demographic and random amplified polymorphic DNA analyses reveal high levels of genetic diversity in a clonal violet. AB - We performed demographic and molecular investigations on woodland populations of the clonal herb Viola riviniana in central Germany. We investigated the pattern of seedling recruitment, the amount of genotypic (clonal) variation and the partitioning of genetic variation among and within populations. Our demographic study was carried out in six violet populations of different ages and habitat conditions. It revealed that repeated seedling recruitment takes place in all of these populations, and that clonal propagation is accompanied by high ramet mortality. Our molecular investigations were performed on a subset of three of these six violet populations. Random amplified polymorphic DNA analyses using six primers yielded 45 scorable bands that were used to identify multilocus genotypes, i.e. putative clones. Consistent with our demographic results and independent of population age, we found a large genotypic diversity with a mean proportion of distinguishable genotypes of 0.93 and a mean Simpson's diversity index of 0.99. Using AMOVA we found a strong genetic differentiation among these violet populations with a PhiST value of 0.41. We suggest that a high selfing rate, limited gene flow due to short seed dispersal distances and drift due to founder effects are responsible for this pattern. Although Viola riviniana is a clonal plant, traits associated with sexual reproduction rather than clonality per se are moulding the pattern of genetic variation in this species. PMID- 11472549 TI - Genetic differentiation of a European caddisfly: past and present gene flow among fragmented larval habitats. AB - We describe the genetic structure of a freshwater insect species and interpret it in terms of present-day population dynamics and possible postglacial colonization history. The sampling regime represented a large area of the species range in northwest Europe, particularly focusing on Britain, a region relatively neglected in molecular population genetic studies. Plectrocnemia conspersa generally showed low levels of genetic variation across the sampled populations (Nei's D = 0.0138) and subdivision was unrelated to the pattern of the drainage network. However, the results do suggest that populations across the region are not at equilibrium and that British populations still show effects of the recolonization of the species following the last glacial maximum. Levels of genetic diversity were lower in Britain than in mainland Europe. Two-dimensional scaling showed genetic differentiation between major regions and the pattern of genetic diversity indicates a more recent origin of populations in the north and west of the area compared with the south and east. We argue that, despite the highly fragmented larval habitat, dispersal over tens of kilometres is frequent. Over longer distances, however, P. conspersa does still show evidence of founder effects and postglacial range expansion into Britain. PMID- 11472550 TI - Quantitative polymerase chain reaction analysis of DNA from noninvasive samples for accurate microsatellite genotyping of wild chimpanzees (Pan troglodytes verus). AB - Noninvasive samples are useful for molecular genetic analyses of wild animal populations. However, the low DNA content of such samples makes DNA amplification difficult, and there is the potential for erroneous results when one of two alleles at heterozygous microsatellite loci fails to be amplified. In this study we describe an assay designed to measure the amount of amplifiable nuclear DNA in low DNA concentration extracts from noninvasive samples. We describe the range of DNA amounts obtained from chimpanzee faeces and shed hair samples and formulate a new efficient approach for accurate microsatellite genotyping. Prescreening of extracts for DNA quantity is recommended for sorting of samples for likely success and reliability. Repetition of results remains extensive for analysis of microsatellite amplifications beginning from low starting amounts of DNA, but is reduced for those with higher DNA content. PMID- 11472551 TI - Population-scale laboratory studies of the effect of transgenic plants on nontarget insects. AB - Studies of the effects of insect-resistant transgenic plants on beneficial insects have, to date, concentrated mainly on either small-scale "worst case scenario" laboratory experiments or on field trials. We present a laboratory method using large population cages that represent an intermediate experimental scale, allowing the study of ecological and behavioural interactions between transgenic plants, pests and their natural enemies under more controlled conditions than is possible in the field. Previous studies have also concentrated on natural enemies of lepidopteran and coleopteran target pests. However, natural enemies of other pests, which are not controlled by the transgenic plants, are also potentially exposed to the transgene product when feeding on hosts. The reduction in the use of insecticides on transgenic crops could lead to increasing problems with such nontarget pests, normally controlled by sprays, especially if there are any negative effects of the transgenic plant on their natural enemies. This study tested two lines of insect-resistant transgenic oilseed rape (Brassica napus) for side-effects on the hymenopteran parasitoid Diaeretiella rapae and its aphid host, Myzus persicae. One transgenic line expressed the delta-endotoxin Cry1Ac from Bacillus thuringiensis (Bt) and a second expressed the proteinase inhibitor oryzacystatin I (OC-I) from rice. These transgenic plant lines were developed to provide resistance to lepidopteran and coleopteran pests, respectively. No detrimental effects of the transgenic oilseed rape lines on the ability of the parasitoid to control aphid populations were observed. Adult parasitoid emergence and sex ratio were also not consistently altered on the transgenic oilseed rape lines compared with the wild-type lines. PMID- 11472552 TI - Prospects and challenges in lymphatic filariasis. PMID- 11472553 TI - Immunological genomics of Brugia malayi: filarial genes implicated in immune evasion and protective immunity. AB - Filarial nematodes are metazoan parasites with genome sizes of> 100 million base pairs, probably encoding 15 000-20 000 genes. Within this considerable gene complement, it seems likely that filariae have evolved a spectrum of immune evasion products which underpin their ability to live for many years within the human host. Moreover, no suitable vaccine currently exists for human filarial diseases, and few markers have yet been established for diagnostic use. In this review, we bring together biochemical and immunological data on prominent filarial proteins with the exciting new information provided by the Filarial Genome Project's expressed sequence tag (EST) database. In this discussion, we focus on those genes with the highest immunological profile, such as inhibitors of host enzymes, cytokine homologues and stage-specific surface proteins, as well as products associated with the mosquito-borne infective larva which offer the best opportunity for an anti-filarial vaccine. These gene products provide a fascinating glimpse of the molecular repertoire which helminth parasites have evolved to manipulate and evade the mammalian immune response. PMID- 11472554 TI - Divergent roles for macrophages in lymphatic filariasis. AB - Macrophages have long been recognized as important cells associated with filarial infection but their function as effectors and/or suppressors has not been elucidated. Recent advances in our understanding of the role that macrophages may play in lymphatic filariasis have come from in vitro studies and mouse models of filarial infection. Based on these new findings, we hypothesize that while dead or dying worms induce the 'classical' activation of macrophages and a subsequent pro-inflammatory response, live and healthy worms secrete products that induce type 2 cytokines and the differentiation of 'alternatively' activated macrophages that downregulate an inflammatory response. Thus, the balance between the 'classical' and 'alternative' activation pathways of macrophages could be an important factor in inflammatory pathology associated with filariasis. PMID- 11472555 TI - Lymphatic filariasis: parallels between the immunology of infection in humans and mice. AB - Mouse models of Brugia infection have provided much useful quantitative and qualitative information on the immune response elicited by different life cycle stages of filarial worms. Many parallels exist between the immune response in the mouse and the infected human and in this review we highlight areas of topical interest, including the induction of specific cytokine responses and their role in immunomodulation and protective immunity. These studies have reinforced the concept that different life cycle stages of filarial parasites each have their own mechanism of modulating responses so that potentially inflammatory IFN-gamma responses are downregulated. While the precise mechanisms of protective immunity remain to be defined, studies in the mouse have suggested novel pathways, including a possible role for granulocytes. PMID- 11472556 TI - Transmission intensity and human immune responses to lymphatic filariasis. AB - Our understanding of how the host immune response influences the risk of developing disease has changed dramatically over the past decade. Previously, the spectrum of disease associated with lymphatic filariasis was largely attributed to the nature of the host immune response. Now, we appreciate that the duration and intensity of infection and possibly the direct influence of parasite-derived molecules also determine the risk of disease. Individuals chronically infected with lymphatic filariasis generally have an impaired lymphocyte proliferation response to filarial antigens and favour Th2-type cytokine responses. This ability to down-modulate the host immune response may help protect the host from disease. Defects in antigen-presenting cell (APC) function appear to participate in this acquired immune hyporesponsiveness, although the mechanisms as to how this occurs are poorly understood. Here, we present evidence that repeated exposure to infective stage larvae and their secreted products may stimulate basophils and mast cells to related products that may impair APC function. PMID- 11472557 TI - Transmission intensity and the immunoepidemiology of bancroftian filariasis in East Africa. AB - Previous attempts to determine the interactions between filariasis transmission intensity, infection and chronic disease have been limited by a lack of a theoretical framework that allows the explicit examination of mechanisms that may link these variables at the community level. Here, we show how deterministic mathematical models, in conjunction with analyses of standardized field data from communities with varying parasite transmission intensities, can provide a particularly powerful framework for investigating this topic. These models were based on adult worm population dynamics, worm initiated chronic disease and two major forms of acquired immunity (larval- versus adult-worm generated) explicitly linked to community transmission intensity as measured by the Annual Transmission Potential (ATP). They were then fitted to data from low, moderate and moderately high transmission communities from East Africa to determine the mechanistic relationships between transmission, infection and observed filarial morbidity. The results indicate a profound effect of transmission intensity on patent infection and chronic disease, and on the generation and impact of immunity on these variables. For infection, the analysis indicates that in areas of higher parasite transmission, community-specific microfilarial rates may increase proportionately with transmission intensity until moderated by the generation of herd immunity. This supports recent suggestions that acquired immunity in filariasis is transmission driven and may be significant only in areas of high transmission. In East Africa, this transmission threshold is likely to be higher than an ATP of at least 100. A new finding from the analysis of the disease data is that per capita worm pathogenicity could increase with transmission intensity such that the prevalences of both hydrocele and lymphoedema, even without immunopathological involvement, may increase disproportionately with transmission intensity. For lymphoedema, this rise may be further accelerated with the onset of immunopathology. An intriguing finding is that there may be at least two types of immunity operating in filariasis: one implicated in anti-infection immunity and generated by past experience of adult worms, the other involved in immune mediated pathology and based on cumulative experience of infective larvae. If confirmed, these findings have important implications for the new global initiative to achieve control of this disease. PMID- 11472558 TI - Regulation of the immune response in lymphatic filariasis: perspectives on acute and chronic infection with Wuchereria bancrofti in South India. AB - Delineating the immune responses in lymphatic filariasis has been complicated not only by the rapidly expanding knowledge of new immunological mediators and effortors, but also by new methodologies (in particular, circulating filarial antigen detection) for defining and categorizing filarial-infected individuals. By using assays for circulating antigen in the sera collected as part of the many immunological studies performed on individuals in a Wuchereria bancrofti-endemic region of South India, we have attempted to explore the influence of patency on the antigen-driven proliferative and cytokine responses seen in peripheral blood mononuclear cells of individuals with varying clinical manifestations of lymphatic filarial infection. Moreover, we have provided perspectives on the differences between acute and chronic infection with W. bancrofti and suggested mechanisms that may underly the modulation of the immune response as patency occurs. PMID- 11472559 TI - Wolbachia bacteria in filarial immunity and disease. AB - Lymphatic filarial nematodes are infected with endosymbiotic Wolbachia bacteria. Lipopolysaccharide from these bacteria is the major activator of innate inflammatory responses induced directly by the parasite. Here, we propose a mechanism by which Wolbachia initiates acute inflammatory responses associated with death of parasites, leading to acute filarial lymphangitis and adverse reactions to antifilarial chemotherapy. We also speculate that repeated exposure to acute inflammatory responses and the chronic release of bacteria, results in damage to infected lymphatics and desensitization of the innate immune system. These events will result in an increased susceptibility to opportunistic infections, which cause acute dermatolymphangitis associated with lymphoedema and elephantiasis. The recognition of the contribution of endosymbiotic bacteria to filarial disease could be exploited for clinical intervention by the targeting of bacteria with antibiotics in an attempt to reduce the development of filarial pathology. PMID- 11472560 TI - Regulatory effects of senescence marker protein 30 on the proliferation of hepatocytes. AB - Senescence marker protein 30 (SMP 30) is preferentially expressed in the liver. One of its remarkable functions is the protection of cells against various injuries by enhancement of membrane calcium-pump activity. We analyzed the role of SMP 30 in hepatocyte proliferation. SMP 30 expression was decreased initially, then increased along with hepatic regeneration, after carbon tetrachloride (CCl4) administration. SMP 30 expression was decreased in the necrotic phase and then gradually increased. Its increase was slightly delayed just after the mitotic phase. These results lead us to speculate that mitoses of hepatic cells induce enhanced SMP 30 expression. In contrast, administration of lead nitrate (LN) as a hepatic mitogen induced a more stable increase of SMP 30 expression. To estimate the effect of SMP 30 on cell proliferation, we evaluated hepatic mitosis in wild type and SMP 30-deficient knockout (KO) mice after CCl4 administration. We found an increase in mitotic numbers in hepatocytes of KO mice. This result suggests that SMP 30 has a suppressive effect on cell proliferation. Suppressive activity of SMP 30 cDNA was shown in cultured hepatoblastic cells. Our results suggest that SMP 30 performs a regulatory function in liver regeneration. PMID- 11472561 TI - Apoptosis as an independent prognostic indicator in squamous cell carcinoma of the esophagus. AB - Apoptosis plays a crucial role in determining net cell proliferation and cell turnover in various tumors. The rate of apoptosis in tumor cells has been reported to be a useful prognostic indicator in colorectal carcinoma. We examined apoptosis in 72 specimens of esophageal squamous cell carcinoma, by the terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) digoxigenin-nick end labeling (TUNEL) method. We examined correlation of apoptosis with outcome, clinicopathological features, and expression of the apoptosis-related proteins p53 and Bcl-2. The percentage of apoptotic cells, or apoptotic index (AI), ranged from 0.8 to 9.4 (mean: 3.47; SD: 2.02). Overall, 5 year survival of patients with high AI (AI > or = 5.0; n = 18) tumors was significantly higher than that of patients with low AI tumors (AI < 5.0; n = 58; 76.9% versus 44.9%; P = 0.042). AI did not correlate significantly with the clinicopathological features of patient age and sex, depth of tumor and histological differentiation, lymph node metastasis, lymphatic invasion, or venous invasion. In p53-negative tumors, the AI was significantly higher than in p53-positive tumors. We concluded that AI may be a useful prognostic indicator in esophageal squamous cell carcinoma following curative surgery, and that apoptosis in this tumor is related to relative underexpression of p53 protein. PMID- 11472562 TI - Multiple minute carcinoids in type A gastritis: attempt at 3-D reconstruction. AB - In type A gastritis, the numbers of endocrine cell micronests (ECM) and carcinoids increase through the trophic action of gastrin. This study examined the characteristics and growth of carcinoids in type A gastritis. A total of 395 lesions in five surgically removed stomachs with type A gastritis were investigated, in terms of number, size, distribution and histological appearance, to clarify the tumorigenesis and progression of carcinoids. 3-D reconstruction using serial paraffin sections was used to study carcinoid progression. Our findings suggest that in type A gastritis, carcinoids arise in areas where minute carcinoids are present at a high density. They also suggest that early stage carcinoids not only become large expansively, but also develop in a very complex manner, by maintaining contact with surrounding minute carcinoids. PMID- 11472563 TI - Cytokeratin subtypes in biliary atresia: immunohistochemical study. AB - The etiology of biliary atresia (BA) remains unknown, but ductal-plate malformation and insufficient ductal-plate remodeling have been suggested to play important roles, so it is beneficial to examine the maturation and differentiation of bile ducts in BA. Different epithelial types are characterized by the expression of specific cytokeratin (CK) subtypes. CK can therefore serve as a 'lineage marker' of epithelial cells. CK subtypes have not been previously examined in BA. In this study, we examined the maturation of bile-duct cells in BA (n = 45) using immunohistochemistry of CK subtypes, with mouse monoclonal antibodies to CAM5.2, and CK subtypes 7, 8, 13, 14, 17, 19 and 20. We then compared these findings with pediatric non-BA (n = 11) and fetal (n = 21) liver. We semiquantitatively evaluated the findings using a H score method. In the fetal liver, immunoreactivity for CAM5.2, CK-7, CK-8 and CK-19 was detected in bile duct cells, and CAM5.2 and CK-8 immunoreactivity was also detected in hepatocytes. The distribution of these CK subtypes was the same in fetal, pediatric non-BA and BA liver. However, CK-7 immunoreactivity was markedly weaker in bile ducts of fetal (H scores: ductal plate 0 +/- 0; remodeling 9.5 +/- 40.3; remodeled 37.3 +/- 60.8) and BA (H score: 200.9 +/- 55.3) liver compared to non BA liver (H score: 251.1 +/- 33.5). In addition, CK-20 was detected in the bile ducts of the fetal and BA liver, but not in non-BA liver. These findings suggest that the expression patterns of CK subtypes in bile-duct cells in BA are similar to that in developing bile-duct cells, which is indicative of bile-duct cell immaturity. PMID- 11472564 TI - p53 expression in gastrointestinal stromal tumors. AB - The understanding of gastrointestinal stromal tumors (GIST) is complex because of their divergent differentiation and unpredictable behavior. However, our understanding is becoming clearer, despite some cases of tumors which are exceptions from the typical cases. Tumor size, mitotic rate and, to a lesser degree, location, are the most important predictive parameters for the behavior of GIST. In this study, expression of p53 protein was evaluated in 15 cases of GIST. Tumors were divided into three groups: (i) benign (mitotic index [MI] < 5/50 high-power fields [HPF] and size < 5 cm); (ii) borderline (MI < 5/50 HPF and size > or = 5 cm); and (iii) malignant (MI > or = 5/50 HPF, irrespective of size). The mean values of p53 expression in the three groups were significantly different (benign, 10.6%; borderline, 33.8%; and malignant, 71%). The conclusion of the present study is that p53 overexpression correlates well with the malignant potential of GIST. PMID- 11472565 TI - Gastrointestinal stromal tumors and KIT-positive mesenchymal cells in the omentum. AB - Gastrointestinal stromal tumor (GIST) is currently considered to be derived from the interstitial cells of Cajal (ICC). To test the hypothesis that omental mesenchymal tumor is also a type of GIST, we evaluated the expression of specific molecules in GIST, and c-kit gene mutation in omental mesenchymal tumors, and we identified a possible counterpart of ICC in the omentum. Immunohistochemically, all of the omental mesenchymal tumors (n = 5) were positive for both KIT and CD34, and three of the five tumors were also positive for an embryonic form of smooth-muscle myosin heavy chain (SMemb). Polymerase chain reaction-single-strand conformational polymorphism analysis (PCR-SSCP) and direct sequencing revealed mutations in c-kit gene exon 11 in all five tumors. As for the ICC counterparts in the omentum, there were some KIT-positive mesenchymal cells resembling ICC at the surface of the omentum. Double fluorescence immunostaining, using anti-KIT polyclonal antibodies and monoclonal antibodies against other molecules, demonstrated that KIT-, CD34- and SMemb-positive cells were present just beneath the mesothelial cells of the omentum. These results show that omental mesenchymal tumor corresponds to GIST of the omentum, and that KIT-positive bipolar mesenchymal cells may be a counterpart of ICC in the gastrointestinal tract. Identification of a new type of KIT-positive mesenchymal cell in the omentum may lead to the discovery of a new physiological role for this organ. PMID- 11472566 TI - Expression of intercellular adhesion molecules in epithelioid sarcoma and malignant rhabdoid tumor. AB - We clinicopathologically evaluated 31 cases of epithelioid sarcoma (ES; 25 'classical' type and six 'proximal variant' type) and six cases of malignant rhabdoid tumor (MRT; three extrarenal and three renal). We also did immunohistochemical studies on 12 classical and three proximal variant cases of ES, and six cases of MRT, to clarify the differences in biological behavior in these tumors. E-cadherin, beta-catenin and CD34 expression was evaluated. We also carried out mutational analysis of exon 3 of the beta-catenin gene by polymerase chain reaction-single-strand conformation polymorphism analysis. In ES, the 5- and 10-year survival rates were 71.1 and 55.3%, respectively. A high mitotic rate (>15/10 high-power fields) was significantly correlated with a poor overall survival rate in ES (P = 0.0248). E-cadherin expression was observed in nine cases (69.2%) of ES and in four cases (66.7%) of MRT. Most of these tumors showed aberrant E-cadherin expression. Seven cases (46.7%) of ES were positive for CD34, although none of the cases of MRT were CD34 positive. Eleven cases (73.3%) of ES were positive for beta-catenin, which was localized to the cellular membrane, whereas all of the cases of MRT were beta-catenin negative. Mutational analysis for the beta-catenin gene was done in nine cases of ES and six cases of MRT, however, genetic alteration was not found. From our results, we conclude that beta-catenin membranous expression could be a useful marker for distinguishing ES, including the proximal variant, from MRT. PMID- 11472567 TI - beta-Catenin gene mutation in human hair follicle-related tumors. AB - beta-Catenin, a multifunctional protein related to the adherens junction and to signal transduction, is a key molecule of cell proliferation, and it is central to epithelial architecture, regulating the polarity of cells and tissues. beta Catenin stabilization may play a key role in epidermal signaling leading to hair development, and its aberrant activation may be implicated in formation of hair tumors. Several investigators have shown that pilomatricomas are frequently associated with beta-catenin mutation. In this study, we confirmed beta-catenin gene (CTNNB1) mutation in human pilomatricomas (100% frequency) from which adequate DNA could be obtained for gene analysis. A novel mutation, D32N, was found in one case of pilomatricoma. A preliminary immunohistological study revealed prominent beta-catenin staining in basophilic cells of pilomatricomas, especially in nuclei. Benign tumors which were considered to be derived from hair matrix or hair follicles, and other benign skin tumors, were also investigated. beta-Catenin mutations were not detected in any of the these tumors. These results seem to indicate that hair matrix cells are key players in hair development. Investigation into gene abnormalities of hair-follicle tumors may elucidate the cause of their neoplastic transformation, and may provide a suggestion for the mechanism of hair development. PMID- 11472568 TI - Adenocarcinoma arising in gastric heterotopic pancreas: clinicopathological and immunohistochemical study with genetic analysis of a case. AB - Heterotopic pancreas in the stomach is a relatively common congenital condition, but the risk of malignant transformation is extremely low. In this study, we describe a case of adenocarcinoma arising from a gastric heterotopic pancreas and we consider its morphological and immunohistochemical features and genetic analysis, in order to examine its histogenesis. This unusual sequela was seen in a 57-year-old woman. Image studies showed a protruding lesion with a central ulcer located in the lesser curvature from the angle to the body of the stomach. A biopsy specimen confirmed this lesion as adenocarcinoma before total gastrectomy. The tumor showed mixed patterns of solid neoplastic-cell proliferation and moderately differentiated glandular structures, and also showed transitional lesions to obvious malignancy, that is, dysplasia, or adenocarcinoma in situ. Neoplastic cells had positive immunoreactivity for carbohydrate antigen (CA) 19-9, mucin (MUC) 1, and insulin, and the mutant allele-specific amplification method revealed a point mutation at K-ras codon 12 (GGT [Gly]-->GAT [Asp]), which is the most common mutational change observed in patients with pancreatic carcinoma. The features of the present case provide clear evidence that this tumor originated from heterotopic pancreatic tissue rather than from gastric epithelium. PMID- 11472569 TI - Carcinoma of the urinary bladder with a lymphomatous appearance. AB - We present the first report of bladder carcinoma that demonstrates a mixture of two distinct histological patterns resembling malignant lymphoma. The patient was a 79-year-old man. One of the histological patterns was a diffuse growth of monomorphic carcinoma cells, and the other was a dense lymphoplasmacytic infiltrate, obscuring the carcinoma. The tumor cells showing both patterns expressed cytokeratin and epithelial membrane antigen, but not lymphoid markers. Careful immunohistochemical evaluation should be done when diagnosing urinary bladder carcinomas resembling lymphomas (other than primary lymphomas). PMID- 11472570 TI - Intrarenal mature cystic teratoma associated with renal dysplasia: case report and literature review. AB - We report a case of intrarenal teratoma in a 6-year-old boy. Two years before his operation, multicystic masses had been found in the left side of his abdomen. In the operation, three main cystic masses were located in the upper and lower poles of the left kidney, which were removed in pieces. Histologically, the cyst wall was lined mainly with keratinizing squamous epithelium with hair follicles, shafts and sebaceous glands. The adjacent renal parenchyma showed atrophy, with partially dysplastic and angiomyolipoma-like lesions. Based on these findings, the lesion was diagnosed as mature cystic teratoma of dermoid cyst type. Extragonadal teratoma occurs predominantly along the median line of the body. Intrarenal teratoma is extremely rare; however, it should be distinguished from teratoid Wilms' tumor and other renal cystic lesions. PMID- 11472571 TI - Pigmented neurofibroma: report of two cases and literature review. AB - Two cases of pigmented neurofibroma of the skin are reported. In case 1, the tumor was removed from the back of a 55-year-old man with no associated neurofibromatosis. In case 2, the tumor was removed from the abdominal wall of a 21-year-old woman with neurofibromatosis. Both tumors consisted of benign, short spindle cells and multiple foci of scattered melanin-laden cells. In case 1, the spindle cells were arranged in a storiform pattern, resembling features of dermatofibrosarcoma protuberans. Immunohistochemically, the spindle cells of both cases were demonstrated to be positive for S-100 protein and CD34. The melanin laden cells stained positively for HMB-45. This report describes an additional two cases of pigmented neurofibroma that conform to the new diagnostic criteria for this disease. PMID- 11472572 TI - Neurofibromatosis type 1-associated unusual pleomorphic astrocytoma displaying continual malignant progression. AB - Patients with neurofibromatosis type 1 (NF1) often have gliomas as a complication, most of which are benign pilocytic astrocytomas which have arisen in optic pathways. In the present case, a 17-year-old girl (at death) with stigmata of NF1, initially had a bulky tumor mass in the left thalamus, developing into the lateral ventricle, at 13 years of age. Partially resected tissue samples showed pleomorphic astrocytoma with abundant xanthoma cells and degenerative structures such as Rosenthal fibers (RF) and eosinophilic granular bodies. Fine eosinophilic granules identical to RF, both immunophenotypically and ultrastructurally, were also seen. The residual tumor was subtotally resected 6 months later, and the tumor histology was essentially similar as before, accompanying the regenerative structures; this was believed to be a good prognostic indicator. However, several anaplastic features such as mitosis, necrosis and vascular proliferation appeared even in areas rich in the regenerative structures. After a 2-year, disease-free interval, multiple tumor relapse occurred in June 1997. Partially resected tumor tissues were composed of monotonous small anaplastic cells with prominent proliferative activity. Surprisingly, the tumor cells had retained eosinophilic granules within the cell bodies. Postoperative chemotherapy with procarbazine, MCNU and vincristine (PCV) suppressed the residual tumor dramatically, but the regrowing tumor finally became uncontrollable, leading to the patient's death. TP53 mutation was not detected, while p27 immunopositivity was constantly high during malignant progression, suggesting acquisition of proliferative activity to overcome p53 and p27 inhibitory functions. A review of previously published reports failed to reveal any cases of this type. PMID- 11472573 TI - End-tidal carbon monoxide is predictive for neonatal non-hemolytic hyperbilirubinemia. AB - BACKGROUND: The aim of the present study was to evaluate the value of an end tidal carbon monoxide corrected for inhaled carbon monoxide concentration (ETCOc) at the early neonatal period. The value would be useful for predicting subsequent hyperbilirubinemia in non-hemolytic full-term infants. METHODS: The ETCOc levels were measured every 6 h during the first 72 h of life in healthy, full-term, non hemolytic, newborn Japanese infants using a breath carbon monoxide analyzer. The ETCOc levels in hyperbilirubinemic infants were compared with those in non hyperbilirubinemic infants. Hyperbilirubinemia was defined as the level of the peak total serum bilirubin concentration (TBC) greater than or equal to 257 micromol/L (15.0 mg/dL). The ETCOc measurement for predicting subsequent hyperbilirubinemia was evaluated with a receiver-operating characteristic (ROC) curve. RESULTS: Fifty-one infants were enrolled in the study. Seven of the 51 infants developed hyperbilirubinemia. The ETCOc levels in non-hyperbilirubinemic infants were decreased in the first 72 h after birth. However, those in the hyperbilirubinemic infants were not decreased significantly, and were higher than those in non-hyperbilirubinemic infants at 42, 48, 54 and 66 h of age. The ETCOc level at 42 h of age was the most predictive of subsequent hyperbilirubinemia by ROC analysis. At the cut-off level of 1.8 microL/L (p.p.m.), the sensitivity, the specificity, the positive predictive value and negative predictive value were 86, 80, 40 and 97%, respectively. CONCLUSION: Increased ETCOc level in the early neonatal period is associated with subsequent hyperbilirubinemia, even in infants without hemolytic disease. The ETCOc measurement may be useful as a screening test for predicting hyperbilirubinemia without hemolytic diseases. PMID- 11472574 TI - Functional immaturity of cord blood monocytes as detected by impaired response to hepatocyte growth factor. AB - BACKGROUND: Monocytes as antigen-presenting cells play an important role in host defense and transplantation. However, there are little reports on cord blood monocytes, and the role of monocytes in cord blood transplantation is largely unknown. METHODS AND RESULTS: There are several cytokines affecting monocyte function. These include interferon-gamma, interleukin-4, interleukin-10, granulocyte macrophage-colony stimulating factor and hepatocyte growth factor (HGF). We investigated the effect of these cytokines on antigen-presenting capacity (APC) of cord and adult blood monocytes. Using either mononuclear cells or purified CD4+ T cells as responder cells, HGF enhanced APC of adult monocytes most effectively among these cytokines. In contrast, cord blood monocytes failed to respond to HGF. As HLA, costimulatory and adhesion molecules may affect APC function, we examined these antigens of monocytes following HGF stimulation. The HGF upregulated integrin alpha5 subunit (CD49e) and intercellular adhesion molecule-1 (CD54) was expressed in adult blood monocytes, but not in cord blood. In kinetic studies, HGF downregulated c-met protein/HGF receptor expression of adult monocytes in lower concentrations and at shorter incubation time as compared with that of cord blood. CONCLUSIONS: The results suggest that impaired response of cord blood monocytes to HGF may be responsible, in large part, for their functional immaturity. PMID- 11472575 TI - Spontaneous labor increases nitric oxide synthesis during the early neonatal period. AB - BACKGROUND: This paper aimed to assess the influence of spontaneous labor upon endogenous nitric oxide (NO) and endothelin 1 (ET-1) during transition to extrauterine life. METHODS: The serum levels of NO metabolites (the sum of nitrites and nitrates (NOx)) and the plasma level of ET-1 were determined in 53 healthy full-term infants (spontaneous labor group; n=40, cesarean delivery group; n=13). In both groups, blood samples were obtained from a cord vein at birth and from a peripheral vein at 5 days of age. RESULTS: The differences in serum NOx concentrations between the spontaneous labor group and the elective cesarean group were not significant at birth. By the age of 5 days, serum NOx concentrations had risen significantly in the spontaneous labor group to become significantly higher in the elective cesarean group. CONCLUSION: It is speculated that spontaneous labor might enhance endogenous NO synthesis at 5 days of age. PMID- 11472576 TI - Nitric oxide inhalation and nitric oxide synthase inhibitor supplement for endotoxin-induced hypotension. AB - BACKGROUND: This study was performed to determine whether a combined therapy of nitric oxide (NO) inhalation and nitric oxide synthase (NOS) inhibitor is effective in experimental animals with endotoxin-induced refractive hypotension accompanied by pulmonary hypertension. METHODS: Escherichia coli lipopolysaccharide (1 mg/kg) was administered to 10 newborn piglets to induce endotoxemia. The experiment then began 60 min later, when the systemic arterial pressure dropped. The inhalation of 20 p.p.m. NO at 60 and 120 min of endotoxemia created a control group. Another group was also administered N w-nitro-L-arginine (L-NNA; 5 mg) after the first NO inhalation at 60 min of endotoxemia (the L-NNA group). Pulmonary arterial pressure, systemic arterial pressure and cardiac output were measured and compared among the groups. RESULTS: Three of the 5 piglets in the control group died of hypotensive shock, while in the L-NNA group the systemic arterial pressure recovered to pre-endotoxin administration levels. The L-NNA group produced a further increase in pulmonary arterial pressure against which NO inhalation was effective. CONCLUSION: Nitric oxide inhalation alone carries a potential risk of further lowering systemic arterial pressure in a piglet with hypotension induced by endotoxin, whereas the combined therapy resulted in the recovery of the blood pressure to pre-endotoxin levels. The combined therapy was simultaneously effective against pulmonary hypertension. PMID- 11472577 TI - Toxocara seroprevalence and childhood asthma among Malaysian children. AB - BACKGROUND: The larva of Toxocara spp., a common animal roundworm, may infect non compatible hosts, causing a profound immunological reaction with marked eosinophil and IgE responses, not unlike in atopy. In this study, we determined the seroprevalence of Toxocara exposure in 66 asthmatic and 58 non-asthmatic children. METHODS: Exposure to Toxocara was determined by examining the serum samples of the children for specific IgG antibodies to L2 Toxocara larvae, using a commercially available diagnostic kit. RESULTS: There was no significant difference in the mean age, sex, social class, residence type and presence of domestic pets at home between the two children groups. Children with bronchial asthma were observed to have higher Toxocara seropositivity than that of the non asthmatic controls (21.2 vs 8.6%, P=0.047). CONCLUSION: The observed relationship between exposure to Toxocara infection and bronchial asthma in Malaysian children warrants further evaluation. An understanding of any possible contribution to the pathogenesis of childhood asthma provides a potential avenue for prevention. PMID- 11472578 TI - Serum levels and differential expression of CD44 in childhood leukemia and malignant lymphoma: correlation with prognostic criteria and survival. AB - BACKGROUND: The CD44, a cell surface proteoglycan, participates in a variety of function including tumor dissemination and metastasis. However, there are no available data on the prognostic significance of CD44 expression of tumor tissue correlated with serum sCD44 level in childhood leukemias and lymphomas. METHODS: Serum levels and leukemic cell tumor tissue expression of CD44 were detected in 54 children with acute leukemia and malignant lymphoma. Serum samples were obtained from all patients before treatment and during remission. Twelve age matched healthy children were included as a control group. RESULTS: The serum CD44 levels were significantly higher in patients with Hodgkin's disease (HD), non-Hodgkin's lymphoma (NHL), Burkitt's lymphoma (BL) and acute lymphoblastic leukemia (ALL) than those in the control group. The median values were 1627.0, 1336.0, 1318.5, 1730.4, 902.7 ng/mL, respectively, and P<0.001, P<0.01, P<0.01, P<0.05 in comparisons, respectively. However, there was no significant difference between acute myeloid leukemia (AML) and the control group (median values: 900.3 and 902.7 ng/mL, respectively, P>0.05). Serum sCD44 levels significantly declined in HD, NHL and ALL patients who were in complete remission (median values: 684.0, 573.8 and 1101.1 ng/mL, respectively, P<0.05 in each comparison). Patients with HD had higher levels of serum sCD44 and correlated well with higher erythrocyte sedimentation rate (ESR), B-symptoms and advanced-stage disease (P<0.05, P<0.05 and P<0.01, respectively). Expression of CD44 was significantly high in patients with HD and NHL who were in advanced stages of disease. High serum CD44 level was also associated with high tumor tissue expression of CD44 in patients with HD and BL. In addition, patients with higher levels of serum sCD44, had a poorer outcome and survival than those with lower sCD44 levels in HD and NHL groups. CONCLUSIONS: A high serum sCD44 level and/or tumor tissue expression at diagnosis is associated with poor prognostic criteria and/or unfavorable outcome in childhood leukemias and lymphomas. PMID- 11472579 TI - Skeletal development of achondroplasia: analysis of genotyped patients. AB - BACKGROUND: Achondroplasia is a skeletal dysplasia caused by substitution of arginine for glycine at codon 380 (G380R) mutation of the fibroblast growth factor receptor 3. To date, the developmental course of the phenotype (short stature and skeletal characteristics) has not been clarified in the genotyped population. METHODS: The relationship between age and clinical data (height, arm span and measurements of skeletal radiographs) were statistically analyzed from 27 achondroplasia patients with the G380R genotype. RESULTS: The height standard deviation score had positive correlation and decreased with age, while span-to height ratio did not. Among measurements of skeletal radiographs, the pelvic index, which represents the squared pelvis deformity, were correlated and increased with age. However, interpedicular distance of the first and fourth lumbar vertebrae (L1:L4) ratio as an index for the caudally narrowed pattern of the lumbar spinal canal and fibula-to-tibia ratio for the disproportionally long fibulae were not correlated and did not increase with age. CONCLUSION: In making a clinical diagnosis of achondroplasia in early infancy, it should be noted that short stature and squared pelvis deformity are not prominent. PMID- 11472580 TI - Prognosis of acute poststreptococcal glomerulonephritis (APSGN) is excellent in children, when adequately diagnosed. AB - BACKGROUND: Recently, the prognosis of acute poststreptococcal glomerulonephritis (APSGN) has been reported as improved, compared with the results of previous studies. In an attempt to clarify this, we analyzed the clinical course of patients with APSGN. METHODS: A total of 220 children with acute nephritic syndrome were treated in the affiliated hospitals of our department, between January 1988 and December 1997. Among them, 138 children who were diagnosed with APSGN according to the presence of hematuria, transient hypocomplementemia and evidence of group A beta-hemolytic streptococcal infection, were studied. RESULTS: Serum creatinine and blood urea nitrogen levels at onset were 0.5 +/- 0.2 mg/dL and 20 +/-12 mg/dL, respectively. There were no patients with renal dysfunction (serum creatinine level > or = 1.5 mg/dL), but one patient with nephrotic syndrome. Blood pressure was well controlled in all patients and there were no patients with persistent hypertension. Serum complement levels were normalized within 12 weeks (100%), hematuria disappeared within 4 years (100%) and proteinuria disappeared within 3 years (100%) from the onset. CONCLUSIONS: These data indicate that the prognosis of APSGN during childhood is excellent, when adequately diagnosed and treated. PMID- 11472581 TI - Characteristics of suicide among children and adolescents in Budapest. AB - BACKGROUND: In Europe, the suicide rate among children and adolescents is the highest in Hungary. In the young age group (10-14 and 15-19 years), the suicide mortality rate had not showed a decrease. METHODS: This study examined suicide cases committed by children and adolescents highlighted from the extraordinary death cases in Budapest between 1994 and 1998. The 72 suicide cases, which included 59 males and 13 females, were processed by sex, age, method and time of commitment, and distribution by districts. RESULTS: Leaping off high places or hanging was the highest frequency among the methods of commitment. Accurate data about the frequency, type, time and location of death cases are indispensable to decrease the number of children's and adolescents' suicides. CONCLUSIONS: Studies of epidemiology and risk factors may provide a basis of development of a specific suicide prevention programme including educational, health and welfare elements. PMID- 11472582 TI - Quantitation of human herpesvirus 6 DNA in infant with exanthem subitum by microplate PCR-hybridization assay. AB - BACKGROUND: Quantitative analysis of human herpesvirus 6 (HHV-6) genome is important for monitoring active virus infection. The purpose of our study is to evaluate the reliability of a hybridization-based microtiter plate assay (polymerase chain reaction enzyme-linked immunosorbent assay (PCR ELISA)) for quantifying the virus genome. METHODS: Semiquantitative analysis of the virus genome was carried out in 31 (18 male and 13 female) infants with primary HHV-6 infection. If the HHV-6 virus could be isolated from the peripheral blood mononuclear cells (PBMC), the infants were defined as being infected with HHV-6. The PCR ELISA method was used to determine the virus load. A titration of the virus was also carried out in the samples obtained during the acute phase of exanthem subitum. RESULTS: Specificity of the method was demonstrated by a lack of amplification of human herpesvirus 7 and cytomegalovirus DNA. The upper and lower detection limits of the method were 58 and 5800 copies of the virus genome, respectively. The quantity of HHV-6 DNA in the PBMC during the acute phase (879 +/- 975 copies/10(4) PBMC) was significantly higher than during the convalescent phase (54 +/- 76 copies/10(4) PBMC). Furthermore, the virus load in acute phase plasma (53 +/- 75 copies/microL) was also significantly higher than in the convalescent phase samples (2 +/- 9 copies/microL). Virus load in both PBMC and plasma gradually increased after the onset of exanthem subitum until about day 3 to 4 of the illness, but then decreased quickly. However, there was no significant association between virus load and the numbers of infected cells. CONCLUSION: Virus load in both PBMC and plasma gradually increased after the onset of exanthem subitum until about day 3 and day 4 of the illness, respectively, then it decreased quickly. These results indicate that our PCR ELISA system is reliable for monitoring active HHV-6 infection in vivo. PMID- 11472583 TI - Measles seroprevalence in Izmir with special emphasis on measles vaccination policy for Turkey. AB - BACKGROUND: Measles outbreaks seem to occur every 2- to 3-year intervals in Turkey. However, sero-epidemiological studies are limited. Knowing the prevalence of measles susceptibility as measured either by serologic markers of immunity or surveys of vaccination coverage is an important tool to assess the risk for measles outbreaks. METHODS: In order to determine the seroprevalence of measles antibodies among a 1 to 29-year-old population in Izmir (Turkey) and to develop the best vaccination policy for measles, a total of 600 people aged from 1 to 29 were selected for the study with cluster sampling. The information on sociodemographic characteristics, vaccination status and measles history was gathered for each participant. Measles-specific IgG antibodies were screened qualitatively by using microenzyme immune assay for 595 subjects. RESULTS: Of the 595 participants screened for the measles antibodies, 56 (9.4%) were seronegative. The proportion of the susceptible individuals in the age groups of 1-4, 5-9, 10-14, 15-19 and 20-29 was 20.0, 10.4, 6.0, 10.3 and 3.0%, respectively. The logistic regression analysis showed that none of the independent characteristics (sex, socioeconomic status, past measles history, vaccination status) with the exception of age group, was significantly associated with measles seronegativity. CONCLUSION: The optimal measles vaccination policy for Turkey may be to increase vaccination coverage above 90%, to conduct a catch up campaign covering persons aged 1-19, regardless of previous vaccination status. Another factor to consider is to adopt a routine two-dose vaccination, giving the first dose at 12-15 months of age and the second dose at school entry. PMID- 11472584 TI - Investigation of risk factors for penicillin-resistant Streptococcus pneumoniae carriage in Turkish children. AB - BACKGROUND: Nasopharyngeal colonization plays an important role for infections caused by Streptococcus pneumoniae. Emergence of penicillin resistance in this organism has made it difficult to treat pneumococcal infections. The objectives of this study were to investigate the risk factors for nasopharyngeal colonization with S. pneumonia and for nasopharyngeal colonization with penicillin-resistant S. pneumoniae. METHODS: Three hundred children with or without evidence of infection were investigated for various risk factors. Streptococcus pneumoniae isolated from children's nasopharyngeal swabs were examined for penicillin susceptibility. RESULTS: Day-care attendance (odds ratio OR=2.82, P=0.003) and upper respiratory tract infection within the last month (OR=1.83, P=0.02), have been determined to be risk factors for S. pneumoniae carriage. The use of antibiotics within the last 3 months (OR=81.07, P<0.001), the presence of more than five people living in the house of the child (OR=6.63, P=0.03), and having a sibling under 5-years-old (OR=4.60, P=0.03) have been determined to be risk factors for penicillin-resistant S. pneumoniae carriage. CONCLUSION: Some children are inevitably exposed to and colonized with penicillin susceptible or resistant S. pneumoniae. Changes in day-care organizations, better living conditions, and restriction of antibiotic use seems to be useful precautions to prevent the emerging and colonization with penicillin-susceptible or -resistant S. pneumoniae. PMID- 11472585 TI - Plasma and liver carnitine status of children with chronic liver disease and cirrhosis. AB - BACKGROUND: Carnitine is an essential cofactor in the transfer of long-chain fatty acids across the inner mitochondrial membrane for oxidation. As its synthesis is performed in the liver, alterations in carnitine metabolism is expected in liver diseases, especially in cirrhosis. METHODS: In this study, we investigated plasma and liver carnitine concentrations of 68 children with chronic liver disease, 36 of whom had cirrhosis as well. Carnitine level was determined by enzymatic method. RESULTS: Plasma and liver carnitine concentrations were not correlated. Mean plasma carnitine level of cirrhotic children was significantly lower than that of the control group (P<0. 0001). While there was no difference between liver carnitine concentrations of children with chronic liver disease and cirrhosis (P>0.05), mean plasma level of cirrhotics were lower (P<0.05). Plasma carnitine was correlated with albumin, triglyceride and gamma glutamyl transpeptidase (GGT) in patients with chronic liver disease (P<0.05). Liver carnitine was correlated with GGT in cirrhotic patients (P<0.005). Children with malnutrition had higher plasma and liver carnitine levels (P<0.05). The highest plasma and liver carnitine levels were detected in children with biliary atresia and criptogenic cirrhosis, respectively. Both the lowest plasma and liver carnitine levels were detected in Wilson's disease. CONCLUSION: Children with cirrhosis have low plasma carnitine concentrations. This finding is prominent in children with Wilson's disease. As carnitine is an essential factor in lipid metabolism, the carnitine supplementation for patients with cirrhosis in childhood, especially with Wilson's disease, seems to be mandatory. PMID- 11472586 TI - Serum zinc status in chronic hepatitis B and its relationship to liver histology and treatment results. AB - BACKGROUND: It is known that cytotoxic T lymphocytes are responsible for viral clearance in chronic hepatitis B (HBV) infection. Zinc deficiency affects development of acquired immunity by preventing certain functions of T lymphocytes. We investigated the serum zinc levels and the relationship to liver histopathology and response to interferon alpha (IFN-alpha) and lamivudine combination therapy in 28 children with chronic HBV infection. METHODS: A course of IFN-alpha was injected as 5 million U/m2 subcutaneously, thrice a week for 6 months and lamivudine 4 mg/kg per day orally, for 1 year. Normalization of alanine aminotransferase (ALT), loss of HBV DNA, hepatitis B e antigen (HBeAg) seroconversion altogether was considered as end of therapy response (ETR). RESULTS: The ETR was achieved in eight (30.7%) patients. Serum zinc concentrations of 20 healthy children and patients was not significantly different (P>0.05). While pretreatment serum ALT, zinc, histological activity index (HAI) and portal inflammation scores were statistically higher in children who had ETR (P<0.005, P<0.05, P<0.05 and P<0.05, respectively), pretreatment serum HBV DNA was lower (P<0.005). Serum zinc level was correlated with HAI and portal inflammation scores (P<0.01 and P<0.01). CONCLUSION: This study showed the relationship of serum zinc status to liver histopathology and to the ETR and may be a preliminary study leading new studies focusing on zinc status in patients with chronic HBV infection. PMID- 11472587 TI - Denver developmental screening test II for early identification of the infants who will develop major neurological deficit as a sequalea of hypoxic-ischemic encephalopathy. AB - BACKGROUND: The primary aim of this study was to find widely available, inexpensive, and non-invasive parameters for early identification or prediction of the infants with hypoxic-ischemic encephalopathy (HIE) who will have a severe adverse outcome (classified as death or a major neurological deficit). METHODS: Fifty-seven full-term or near-term newborn infants with a diagnosis of HIE were consecutively admitted to the neonatal intensive care unit and studied. Occurrence of seizures during the first 24 h, cranial ultrasonography (US) findings within the first 5 days of life, and Denver developmental screening test II (DDST II) at 6 months of age, were analyzed in relation to mortality and neurological status at 2 years of age. RESULTS: Of the 57 infants, 10 were lost to follow-up. Twenty of the remaining 47 infants had a severe adverse outcome. Among the predictors of severe adverse outcome, occurrence of seizures was found to have a poor predictive accuracy. Cranial US had 100% sensitivity, however with a rather low specificity (55%). However, DDST II at 6 months of age, yielded a very high predictive accuracy (sensitivity=100%, specificity=95%). CONCLUSION: We conclude that DDST II at 6 months of age could be used in predicting severe neurological outcome in infants with HIE. PMID- 11472588 TI - Primary Duhamel-Martin operations in neonates and infants. AB - BACKGROUND: With advances in neonatal anesthetic and surgical care, a safe, one stage, definitive procedure has been possible in Hirschsprung's disease. Since 1996, we have performed this type of operation in the neonatal and early infancy period. We aimed to review our data to state the feasibility of this operation in these age groups. METHODS: At Dr Behcet Uz Children's Hospital, we treated 10 patients with a single stage Duhamel-Martin operation between 1996 and 2000. Of the 10 patients, seven were boys. Six patients were diagnosed in the first week of the neonatal period. We evaluated these 10 patients by means of age, sex, age at diagnoses, operational age, diagnostic tools, properties of operation, complications and results. RESULTS: The patients were all full-term delivery and had a mean birthweight of 3 kg. The presenting clinical features were abdominal distention (100%), constipation (100%) and vomiting (70%). One patient was a Down syndrome patient, while another patient showed familial Hirschsprung's disease. Contrast enemas gave positive results in eight patients. Definitive diagnoses were performed with rectal biopsy specimens. The extension of the disease was rectosigmoid in nine patients and descending colon in one patient. Five patients were in the newborn period at the time of the operation, while the oldest one was 7 months old. In the postoperative period, two children were treated because of early abdominal eventration and evisceration of the wound. Postoperative enterocolitis occurred in two patients. These 10 patients have been followed-up for a period of 3 years, and spontaneous defecation and weight gain was observed in all of the patients. CONCLUSIONS: Our study confirmed the published data that this operation could be performed as an easy and safe procedure in the neonatal and early infancy period. PMID- 11472589 TI - Omega-3 polyunsaturated fatty acid enriched diet attenuates stress-induced lactacidemia in 10-day-old rats. AB - BACKGROUND: Lactacidemia is often seen under stress conditions including septic shock in the newborn. Under stress conditions, plasma catecholamine concentrations are increased and play an important role in lactate metabolism. Our previous study shows that perinatal feeding of omega-3 polyunsaturated fatty acid enriched diet (omega-3PUFA) attenuates lactacidemia of endotoxic shock in 10 day-old rats. In the omega-6 fatty acids series, decosapentanoic acid, two series prostaglandins and four series leukotrienes are synthesized through linoleic acids. As plasma lactate concentration correlates with the outcome of septic shock in the newborn, it is important to understand the effects of omega-3PUFA on lactate metabolism. Thus, we tested the hypothesis that perinatal feeding of omega-3 polyunsaturated fatty acid enriched diet (omega-3PUFA) alters responses to catecholamines and attenuates the stress-induced lactacidemia in 10-day-old rats. METHODS: Ten-day-old rats which perinatally fed omega-3PUFA. Lactacidemia was induced by swimming for 5 min. Ten-day-old rats which perinatally fed omega 6PUFA were controls. Omega-6 fatty acids series are contained in animal fats and corn oil. Adrenergic blockers were used to assess roles of catecholamines in swimming-induced lactacidemia. RESULTS: Swimming increased plasma lactate concentration less (P<0.05) in rats fed omega-3PUFA than rats fed omega-6PUFA. Swimming increased plasma concentrations of glucose and glucagon, cyclic adenosine monophosphate (cAMP) concentration and phosphoenolypruvate carboxykinase mRNA in the liver, and cAMP concentration in the hindlimb muscle more (P<0.05) in rats fed omega-3PUFA than in rats fed omega-6PUFA. Phentolamine and propranolol enhanced swim-induced lactacidemia in the omega-3PUFA group, while they decreased the lactacidemia in the omega-6PUFA group. Propranolol enhanced swimming-induced hyperglycemia in the omega-6PUFA group more than in the omega-3PUFA group. CONCLUSIONS: Omega-3PUFA might increase beta-adrenergic response in the liver and increase gluconeogenesis in response to stress, resulting in decreased lactacidemia. PMID- 11472590 TI - A boy with systemic lupus erythematosus (SLE) suggesting a familial predisposition for SLE. PMID- 11472591 TI - Systemic lupus erythematosus presenting with normocomplementemic urticarial vasculitis in a 4-year-old girl. PMID- 11472592 TI - Henoch-Schonlein purpura-related intestinal perforation: a steroid complication? PMID- 11472593 TI - 11-year-old boy with chronic renal failure and myelodysplastic syndrome. PMID- 11472594 TI - Chronic mercury intoxication simulating pheochromocytoma: effect of captopril on urinary mercury excretion. PMID- 11472595 TI - Congenital retropharyngeal goiter in a newborn as a magnetic resonance imaging finding. PMID- 11472596 TI - IgA nephropathy associated with Nail-Patella syndrome in a 7-year-old girl. PMID- 11472597 TI - Transient temporal lobe changes and a novel mutation in a patient with Menkes disease. PMID- 11472598 TI - Pulmonary infarction and deep venous thrombosis in a 13-year-old boy with Churg Strauss syndrome. PMID- 11472600 TI - Predicting outcome from post-transplant lymphoproliferative disease: a risky business. PMID- 11472601 TI - Pediatric kidney transplantation: historic hallmarks and a personal perspective. AB - Over the last five decades, pediatric kidney transplantation (Tx) has proved to be a viable therapeutic alternative for children with end-stage renal disease. Patient and graft survival rates, as well as long-term quality of life, have improved dramatically during this time, as a result of advances in surgical techniques, immunosuppression, and pre- and post-operative care. The inspired, hard work of multi-disciplinary clinical teams, combined with the determination and courage of the young patients and their families, have fueled the success of pediatric kidney Tx. It is with similar optimism and drive that we face the great challenges of the future, such as maximizing the donor pool and inducing tolerance. PMID- 11472602 TI - Non-adherence in pediatric transplantation: a review of the existing literature. AB - The degree of medical adherence in pediatric solid-organ transplant recipients frequently correlates with the degree of psychological distress, family functioning, and the physiological side-effects of immunosuppressant medications. This article examines the current literature regarding each of these factors and proposes recommendations for increasing the medical compliance among childhood transplant recipients. PMID- 11472603 TI - Post-transplant lymphoproliferative disease in children. AB - Epstein-Barr virus (EBV)-driven post-transplant lymphoproliferative disease (PTLD) is an important cause of morbidity and mortality following transplantation, and it occurs more frequently in children than in adults. Of 22 (5%) children at our institution who developed tissue-proven PTLD 1-60 months (mean 16.5 months) following organ transplant, 11 died: nine of these 22 patients developed PTLD between 1989 and 1993, and seven (78%) died; the remaining 13 developed PTLD between 1994 and 1998, and four (31%) died (p = 0.08). All nine patients who developed PTLD < 6 months after transplant died, but 11 of 13 patients who manifested disease > or = 6 months after transplant survived (p = 0.0002). Ten of 11 (91%) survivors, but only two of eight (25%) children who died, had serologic evidence of EBV infection at the time of PTLD diagnosis (p = 0.04). EBV seroconversion identified patients at risk for developing PTLD, but also characterized patients with sufficient immune function to survive EBV related lymphoid proliferation. In situ hybridization for EBER1 mRNA was diagnostically helpful because it detected EBV in tissue sections of all 20 patients with B-cell PTLD, including those with negative serology. PMID- 11472604 TI - An in vitro skin explant assay as a predictive assay for graft-versus-host disease in a cohort of pediatric transplants. AB - Severe acute graft-versus-host disease (GvHD) remains a serious complication of allogeneic stem cell transplantation. An in vitro skin explant assay was used to predict the occurrence and severity of acute GvHD in a cohort of 30 pediatric patients undergoing human leucocyte antigen (HLA)-matched sibling transplants (20 patients) and matched or one antigen mismatched unrelated donor transplants (10 patients). In the cohort of HLA-matched sibling transplants, the result appeared to reflect the degree of GvHD prophylaxis. The skin explant assay correlated with GvHD outcome in 12 of 20 children, but this did not reach statistical significance (chi-square 0.95, d.f.=1, p=0.32). These results support previous observations. In this present cohort, patients were treated either with cyclosporin A (CsA) monotherapy (n=7) or with CsA plus additional methotrexate (MTX) (n=13). We have previously demonstrated that the skin explant assay was not as predictive in patients receiving CsA plus additional MTX compared to cohorts treated with CsA alone. In the group of patients treated with CsA alone, four of five patients (80%) predicted to develop GvHD developed acute GvHD of grade II or above; of two patients predicted to develop only grade 0-I GvHD, one patient developed no GvHD and the other grade II GvHD. In the CsA plus MTX group, nine patients were predicted to develop GvHD. Five of nine (55%) developed acute GvHD of grade II or above, while three of four with grade 0 or I skin explant assay results developed only grade 0-I GvHD. In a cohort of 10 patients who received unrelated donor transplants, the skin explant assay correlated with GvHD outcome in all 10 patients (Fisher's exact test p=0.008). Hence, the skin explant assay is a pretransplant in vitro GvHD predictive test that predicts the occurrence and severity of acute GvHD in pediatric unrelated donor transplants and to varying degrees, depending on the GvHD prophylaxis protocols, in HLA-matched sibling cohorts. PMID- 11472605 TI - Incidence, impact on survival, and risk factors for multi-organ system failure in children following liver transplantation. AB - Liver transplantation (LTx) in children currently offers long-term survival rates of more than 80%. Many causes for Tx failure have been identified. However, the incidence and impact of multi-organ system failure (MOSF) are, to date, unknown. Therefore, in this study the role of MOSF after LTx in children was investigated with regard to outcome. The data of 114 children (53 girls, 61 boys; median age 4.3 yr) after first LTx were evaluated retrospectively. The definition of MOSF, as used by Wilkinson et al. [Crit Care Med 1986: 14: 271-274], was modified with regard to age-adjusted values. The influence of MOSF on patient survival was investigated by Kaplan-Meier analysis and multivariate regression analysis. Thirty-one of 114 children with orthotopic LTx developed MOSF (involving two or more organs). In total, 18 children died (15.8%) during the hospitalization; 16 of these had MOSF. Mortality related to two-organ failure was 29.4% (n = 5), to three-organ failure 78% (n = 7), and to four-organ failure 80% (n = 4). The highest mortality rates were observed in children with central nervous system (CNS) and cardiovascular failure, leading to a decreased probability of survival of 0.40 (p < 0.0001). Multi-variate analysis showed that CNS and cardiovascular failure were the most important risk factors for survival (p < 0.0001 and 0.056, respectively). Respiratory and renal failure, in univariate analysis, were significant contributors to poor survival, but had no statistically significant influence on outcome in multivariate analysis. Bone marrow insufficiency was found to have no influence on survival in either analysis. In multivariate analysis, the risk of development of MOSF was significantly increased by high numbers of transfused units of fresh-frozen plasma (FFP), the absence of rejection episodes, or a high bilirubin level prior to Tx. Hence, MOSF is a major factor contributing to the death of children early after LTx. CNS and cardiovascular failure carried the highest risk for a poor outcome. Other risk factors associated with the development of MOSF were: numbers of transfused units of FFP, absence of rejection episodes, and a high pre-Tx bilirubin level. PMID- 11472606 TI - Effect of treatment with prostaglandin E1 and N-acetylcysteine on pediatric liver transplant recipients: a single-center study. AB - Prostaglandin E1 (PGE1) and N-acetylcysteine (NAC) have been used as single agents to decrease reperfusion injury and improve outcome after solid-organ transplantation (Tx). We hypothesized that combined treatment with NAC and PGE1 would be safe and reduce reperfusion injury. We therefore carried out a pilot study to assess the safety of this drug combination and gain information regarding the efficacy of treating pediatric liver transplant recipients with NAC and PGE1. The pilot study took the form of an open-label study incorporating 25 pediatric liver transplant recipients (12 children in the treatment group and 13 children as controls). NAC (70 mg/kg) was given intravenously over 1 h following reperfusion and then every 12 h for 6 days. PGE1 (0.4 mg/kg/h) was given as a continuous intravenous infusion for 6 days, starting after the first NAC dose. The primary outcome was the safety of combined treatment with NAC and PGE1. Patient survival, graft survival, allograft rejection within the first 90 days after Tx, peak post-transplant serum alanine aminotransferase (ALT) concentration, post-transplant length of hospitalization, and post-operative complications were secondary outcomes. Post-operative complications occurred at similar rates in both control and treated groups. No complications or adverse events occurred in the treated group as a result of study drugs. The 3-month patient survival rate was 100% for both groups. For the group treated with NAC and PGE1, peak serum ALT was lower and median length of stay was shorter but the differences did not reach statistical significance. The proportion of patients with allograft rejection was not significantly different between the two groups. However, rejection was more severe in the control group than in the treated group. In summary, infusions of NAC and PGE1 were safely administered to pediatric liver transplant recipients. However, a randomized controlled study is needed to determine the efficacy of treatment with NAC and PGE1. PMID- 11472607 TI - Changes in left ventricular mass index in children and adolescents after renal transplantation. AB - Recent reports indicate a high prevalence of left ventricular hypertrophy (LVH) in children on dialysis and after renal transplantation (Tx), as identified by cross-sectional analysis. However, the evolution of LVH in pediatric patients with end-stage renal disease after renal Tx is not well established. To assess changes of left ventricular mass (LVM), we prospectively performed echocardiography in 23 children and adolescents between November 1998 and July 2000. Each patient had an echocardiographic evaluation while on dialysis (for at least 6 weeks) and a follow-up evaluation at least 6 months after successful renal Tx (i.e. with a measured glomerular filtration rate [GFR] of at least 40 mL/min/1.73 m2). The LVM index was estimated by indexing LVM to height(2.7). Sixteen patients had a cadaveric transplant and seven had a live donor transplant; the mean duration between the two studies was 1.9 +/- 1.6 yr; and the mean GFR was 55.0 +/- 21.4 mL/min/1.73 m2. There was no significant difference in the mean values of the LVM index while on dialysis and after renal Tx (43.9 +/- 17.8 g/m2.7 and 39.3 +/- 12.0 g/m2.7, respectively, p = 0.19), or in the prevalence of LVH (52% and 56%, respectively). Interval changes in the LVM index in individual subjects between the two studies were significantly associated with interval changes in indexed systolic (r = 0.42, p = 0.04) and diastolic (r = 0.42, p = 0.05) blood pressures. Interval changes in hemoglobin, blood urea nitrogen (BUN), creatinine, and duration after Tx did not correlate with changes in the LVM index. There was no significant difference in LVM index change according to the type of dialysis, donor source, and the cause of renal failure. In multivariate analysis, the baseline LVM index and changes in indexed SBP were independent predictors for LVM index change after renal Tx. We conclude that LVH persists in children and adolescents after renal Tx. Control of blood pressure might be an important factor in regression or prevention of progression of LVH in these patients. PMID- 11472608 TI - Polyclonal anti-T-cell globulin as part of the preparative regimen for pediatric allogeneic stem-cell transplantation. AB - To prevent graft rejection and graft-versus-host disease (GvHD) after allogeneic stem-cell transplantation (ASCT), 56 children were given polyclonal anti-T-cell globulin (ATG) as part of the conditioning regimen. Of the 56 children in the cohort, 27 had a non-malignant disease and 29 had different hematological malignancies. Eight were in first remission of leukemia and the remainder in later stages. Donors were in 16 cases a human leucocyte antigen (HLA)-identical sibling and in 40 a matched unrelated donor (MUD). The control group comprised 16 patients with an HLA-identical donor; the children in this group were not treated with ATG. Side-effects related to the ATG treatment occurred in 63% of the patients and included fever, chills, headache, dyspnoea, nausea/vomiting, body pain, fall in blood pressure, and transient respiratory arrest. Engraftment occurred in 55 (98%) of the ATG-treated patients at a median of 17 (11-27) days after ASCT. One rejection occurred at 23 days post-SCT. The probabilities of acute graft-versus-host disease (GvHD) of grades II-IV were 6% for patients with an HLA-identical donor, 12% for controls, and 26% for the MUD group. Chronic GvHD occured in 20%, 50%, and 50% of patients in the three groups, respectively. Transplant-related mortality rates at 100 days were 6%, 6%, and 7%, respectively. The 5-yr survival rate was 94% and 81% using sibling donors, with and without ATG respectively, and 53% using unrelated donors (p = 0.002). Disregarding donor type, among the ATG-treated patients 5-yr survival rates were 46% in patients with a malignant disease and 77% in non-malignant disorders. Relapse and relapse free survival rates were 42% and 46%, respectively. Five out of 12 patients who showed an early full donor chimerism in the T-cell lineage developed acute GvHD of grades II-IV, compared to none out of 13 patients being mixed chimeras (p = 0.01). Hence, the use of polyclonal ATG as part of conditioning prior to ASCT in children is safe and the survival rate encouraging. PMID- 11472609 TI - Single-center experience with mycophenolate mofetil in pediatric renal transplant recipients. AB - Mycophenolate mofetil (MMF), a potent and specific inhibitor of guanosine nucleotide synthesis, is a new immunosuppressive drug used to prevent rejection in transplant patients. Extensive data on its utility and efficacy exists in adult patients but there is limited experience in pediatrics. Twenty-four children (14 male, 10 female; 2-19 yr of age), eight of whom had received living related donor (LRD) transplants and 16 of whom had received cadaveric donor (CD) transplants, have been treated with MMF in our institution since September 1996. MMF was administered for a duration ranging from 13 weeks to 38 months, at an average dose of 600 mg/m2 (range: 200-1,000 mg/dose) every 12 h, for a total experience of 304 patient months. MMF capsules were used in 16 patients and/or pediatric suspension in eight. Five patients were switched to MMF from azathioprine as a result of rejection episodes or inability to taper prednisone, between 5 weeks and 3.5 yr post-transplant. All patients received prednisone, cyclosporin A (CsA), and induction therapy with anti-lymphocyte globulin (19 patients), anti-thymocyte globulin (one patient) or daclizumab (four patients). In 12 patients started on MMF at the time of CD transplant, five (42%) had an acute rejection episode. In seven who received a LRD transplant, one (14%) had an acute rejection episode. No patients who were converted to MMF were treated for acute rejection following conversion to MMF. One LRD graft was lost at 19 days following injury to the donor artery at the time of retrieval. At the last follow up, the average creatinine level was 93 micromol/L and average urea level was 8.6 mmol/L. One patient developed epigastric distress. Three patients developed diarrhea/abdominal pain requiring dose adjustment. Five episodes of leukopenia and one episode of thrombocytopenia required dose adjustment. Two patients developed symptomatic cytomegalovirus (CMV) infection, one while on acyclovir prophylaxis. No malignancy has been encountered to date. Hence, MMF can be administered safely to children with good effect and with an acceptable side effect profile. PMID- 11472610 TI - Efficacy and tolerability of interleukin-2 receptor blockade with basiliximab in pediatric renal transplant recipients. AB - Rejection remains a major threat in pediatric renal transplantation (Tx), causing graft failure and increased exposure to drugs. The new chimeric antibody, basiliximab, directed against the alpha-chain of the interleukin-2 receptor (IL 2R), has been shown to be effective in preventing rejection episodes in adult renal transplant recipients. In our single-center experience from Essen, Germany, we evaluated prospectively the efficacy and tolerability of basiliximab, in combination with cyclosporin A (CsA) and prednisone, in 38 unselected pediatric patients. Mean patient age at Tx was 10.1 yr. Twenty-eight children received a cadaveric organ and 10 children received living-related donor grafts. The 1-yr patient survival rate was 100% and the 1-yr graft survival rate was 95% (36/38 patients). No graft was lost as a result of immunological factors, and single rejection episodes were observed in eight patients (21%). Two of these rejections were steroid-resistant and responded to tacrolimus rescue therapy. The rate of infections was not enhanced; overt cytomegalovirus (CMV) disease was observed in two patients only. Malignancies have not been seen to date. The blockade of the alpha-chain of the IL-2R lasted for up to 6 weeks. We conclude that the addition of basiliximab to standard immunosuppression in pediatric renal transplant recipients is well tolerated and results in a low incidence of rejection. The simple mode of application and the lack of side-effects make basiliximab an especially useful adjunct in pediatric patients. PMID- 11472611 TI - Introduction of mycophenolate mofetil and cyclosporin reduction in children with chronic transplant nephropathy. AB - Chronic transplant nephropathy (CTN) is the most important cause of kidney graft dysfunction. Studies in adult populations have reported a beneficial effect of non-nephrotoxic mycophenolate mofetil (MMF) on graft function in this setting. However, few studies were reported in children in this setting. We therefore reviewed the charts/medical records of renal transplanted patients < 18 yr of age at a single center who had switched from azathioprine to MMF as a result of progressive loss in graft function, for which vascular, infectious, and urological causes were excluded. Serum creatinine (SCr) and calculated creatinine clearance were compared prior to and after MMF introduction. Thirteen patients (nine male/four female), followed-up for 59.3 +/- 35.4 months after transplantation, were analyzed. Age at MMF introduction was 14.2 +/- 3.6 yr. In 11 patients a previous biopsy had shown features of CTN and four patients also presented signs of chronic cyclosporin A (CsA) nephrotoxicity. MMF was started at a dose of 1211 +/- 351 mg/day, and the CsA dose was decreased from 6.69 +/- 3.15 mg/kg/day 6 months before MMF to 4.8 +/- 2.3 mg/kg/day at the time of MMF introduction. CsA was withdrawn in four patients. The median (25-75%) SCr value increased from 1.60 mg/dL (range 1.3 to 1.87 mg/dL) 6 months before MMF to 2.2 mg/dL (range 1.87-2.32 mg/dL) when MMF was introduced. Six months after introduction of MMF, the SCr level had decreased to 1.5 mg/dL (range 1.2-1.8 mg/dL) and remained stable until the last follow-up (17.5 +/- 9.2 months after MMF was started). A similar pattern occured with calculated SCr clearance. There were no acute rejections after changes in immunosuppression. The safety of MMF was also analyzed and in only one patient was the drug stopped as a result of intractable diarrhea. These findings suggest that MMF is sufficiently powerful to allow a decrease/withdrawal of CsA without the burden of acute rejection in a pediatric population with CTN. PMID- 11472612 TI - Oxygen in wound healing: more than a nutrient. PMID- 11472613 TI - The validity of the clinical signs and symptoms used to identify localized chronic wound infection. AB - It is uncertain how accurately classic signs of acute infection identify infection in chronic wounds, or if the signs of infection specific to secondary wounds are better indicators of infection in these wounds. The purpose of this study was to examine the validity of the "classic" signs (i.e., pain, erythema, edema, heat, and purulence) and the signs specific to secondary wounds (i.e., serous exudate, delayed healing, discoloration of granulation tissue, friable granulation tissue, pocketing at the base of the wound, foul odor, and wound breakdown). Thirty-six chronic wounds were assessed for these signs and symptoms of infection with interobserver reliability ranging from 0.53 to 1.00. The wounds were then quantitatively cultured, and 11 (31%) were found to be infected. Increasing pain, friable granulation tissue, foul odor, and wound breakdown showed validity based on sensitivity, specificity, discriminatory power, and positive predictive values. The signs specific to secondary wounds were better indicators of chronic wound infection than the classic signs with a mean sensitivity of 0.62 and 0.38, respectively. None of the signs or symptoms was a necessary indicator of infection, but increasing pain and wound breakdown were both sufficient indicators with specificity of 100%. PMID- 11472614 TI - Dermatitis during radiation for vulvar carcinoma: prevention and treatment with granulocyte-macrophage colony-stimulating factor impregnated gauze. AB - The aim of this study was to determine the effectiveness of granulocyte macrophage colony-stimulating factor (GM-CSF) impregnated gauze in preventing or healing radiation-induced dermatitis. Sixty-one patients were irradiated for vulvar carcinoma. Thirty-seven applied steroid cream at irradiated areas throughout radiotherapy (Group A) and 24 patients applied additionally GM-CSF impregnated gauze (40 micrcog/cm2 of skin-irradiated area, twice per day) in addition to the steroid cream, after 20 Gy of irradiation (Group B). The score of skin reactions (P=0.008, chi2 test) and the time interval of radiotherapy interruption (P=0.037, Mann-Whitney U test) were statistically significantly reduced in Group B patients. Multivariate analysis of variance showed for this group not only a significant reduction in the Sum of Gross Dermatitis Scoring (P<0.001, adjusted for Duration of Dermatitis) but also a significant reduction of the healing time (P=0.02, adjusted for Sum of Gross Dermatitis Scoring). The pain grading was less (P=0.014, chi2 test) and pain reduction was noticed sooner after the application of GM-CSF impregnated gauze (P=0.0017, Mann-Whitney U test). Multivariate logistic regression analysis showed that the only significant effect on dermatitis score is due to Body Mass Index (P=0.034) and the application of GM-CSF (P=0.008). GM-CSF impregnated gauze can be effective in preventing and healing radiation-induced dermatitis and in reducing the interruption intervals of radiotherapy for vulvar carcinomas. PMID- 11472615 TI - Increased collagen deposition in an uncomplicated surgical wound compared to a minimal subcutaneous test wound. AB - Little information is currently available concerning the relationship between results obtained in humans from surgical test wounds and results from wound models. Therefore, to evaluate human wound healing parameters, tubings of expanded polytetrafluoroethylene were implanted in a subcutaneous test wound in the arm of 47 volunteers and 20 patients undergoing hernia repair. The surgical patients also had implants left in the surgical wound cavity. After 10 days the deposition of collagen in the tubings as expressed by hydroxyproline content was 30% higher in the surgical wound than in the test wound, p < 0.05. The amount of collagen deposited in the tubing within the surgical wound did not correlate with measurements in the test wound, whereas a significant correlation of proline levels was found between the two sites, p < 0.05. Deposition of proline and total protein in the model was equivalent. In both wound types age negatively correlated with levels of protein, but not collagen. The variability of the results was 40% lower in the subcutaneous test wound than in the surgical wound. There was no significant difference in hydroxyproline deposition between the volunteers and the patients undergoing hernia repair. In patients undergoing minor surgery without signs of compromised healing the expanded polytetrafluoroethylene test wound in the arm reflects the deposition of non collagenous protein, but not collagen, within the surgical wound. PMID- 11472616 TI - Hyaluronic acid in incision wound fluid: a clinical study with the Cellstick device in children. AB - When inserted into a human incision wound, the Cellstick device harvests inflammatory cells and collects wound fluid, reflecting time-related changes in cell populations and in wound fluid composition. Hyaluronic acid has been postulated to be an important factor in scar reduction in wound healing and in scarless fetal wound healing. The aim of this work was to determine the concentration and variation of hyaluronic acid and proportions of wound cells in closed surgical wounds in children at two time points. The Cellstick device was inserted subcutaneously into the wound at the end of an elective inguinal hernia operation on 37 healthy boys, and the devices were removed 3+/-1 or 24+/-3 hours after surgery. Haluronic acid concentration was measured from the wound fluid and a differential count of the wound cells was performed. There was a significant decrease in hyaluronic acid concentration from 3+/-1 to 24+/-3 hours after surgery (p<0.001, Kruskal-Wallis anova). The variance of hyaluronic acid concentration in wound fluid differed between the wounds at the two time points (p<0.01, Levene test for homogeneity of variance). A positive correlation between hyaluronic acid concentration and patient age (r=0.91, p<0.05, Spearman) at 3+/-1 hours post surgery and between HA and wound lymphocytes (r=0.38, p<0.05, Spearman) was also found. We conclude that the hyaluronic acid concentration in wound fluid peaks early in children and decreases significantly by 3 to 24 hours after surgery, and the concentrations in the wound fluid of healthy boys are more variable 3 hours than at 24 hours after surgery. PMID- 11472617 TI - Expression of cyclin-dependent kinase inhibitors, p21cip1 and p27kip1, during wound healing in rats. AB - Wound healing is a physiological process in which growth of cells is stringently regulated. Cell growth is controlled by cell cycle-related proteins in which the cyclin kinase inhibitors cause cell cycle arrest and inhibit proliferation. However, little is known about the expression and the role of cyclin kinase inhibitors during wound healing in vivo. This study was mainly designed to examine the expression of p21cip1 and p27kip1 in excisional wounds of full thickness skin in rats. Concomitant expression of proliferation marker Ki67 was also examined. Proliferation predominantly occurred in the first week after injury, peaking at postwounding day 5. Expression of both p21cip1 and p27kip1 at the gene and protein levels did occur during wound healing and showed an inverse gradient to that of Ki67. Constitutive p27kip1 was expressed throughout wound healing with low levels during the proliferating period of days 3 and 5 and increased levels during post-mitotic and remodeling stages. In contrast, p21cip1 was expressed transiently with detectable levels only between days 7 and 14 by Western blot analysis. Immunohistochemically, epithelial cells, endothelial cells and fibroblasts all could express both p21cip1 and p27kip1. In conclusion, the overall results suggested that p21cip1 and p27kip1 may play a key role in supervising the growth resulting from cell proliferation in tissue repair. PMID- 11472618 TI - Differential expression and regulation of extracellular matrix-associated genes in fetal and neonatal fibroblasts. AB - Adults and neonates heal wounds by a repair process associated with scarring in contrast to scar-free wound healing in the fetus. In the present study, human dermal fetal fibroblasts, representing the scarless phenotype, and neonatal human dermal fibroblasts, representing scar-forming phenotype, were examined for potential differences that might influence the wound healing process. Fetal fibroblasts secreted four- to tenfold more latent transforming growth factor beta1 depending on the cell strains compared. Fetal fibroblasts also produced higher levels of collagen protein and mRNA for most types of collagen (particularly type III) as compared to neonatal cells. Interestingly, mRNA for type V collagen was significantly reduced in fetal cells. Neonatal fibroblasts expressed significantly higher levels of latent transforming growth factor-beta1 binding protein mRNA, in contrast to almost undetectable levels in fetal fibroblasts. By ligand blot analysis, the levels of insulin-like growth factor binding protein-3, a reported mediator of transforming growth factor-beta1 activity, was eightfold higher in neonatal versus fetal fibroblasts. Approximately 20 other mRNAs for various cytokines, matrix molecules and receptors were examined and found to be similar between the two cell types. The phenotypic differences described in this article may represent potentially important mechanisms to explain the differences in the quality of wound repair observed in fetal versus adult/neonatal tissues. PMID- 11472619 TI - Delayed wound healing in aged rats is associated with increased collagen gel remodeling and contraction by skin fibroblasts, not with differences in apoptotic or myofibroblast cell populations. AB - Aging has been anecdotally reported to result in prolonged wound healing. Measurement of punch biopsy wound closure in young (4 month old) and old (36 month old) rats indicated there was a significant delay in wound closure by old rats during the early phase of repair, after which closure rates were equivalent. The delay in granulation tissue accumulation in older animals could involve premature programmed cell death (apoptosis); however, apoptotic fibroblasts in sponge granulation tissue and tissue culture were less abundant in samples from old rats relative to young rats. Myofibroblasts express alpha-smooth muscle actin, and they are believed to be important in wound contraction. There were no significant differences in overall abundance or distribution of alpha-smooth muscle actin containing myofibroblasts in granulation tissue and in cultured granulation tissue fibroblasts regardless of the age of the donor rat. The spatial distribution of myofibroblasts and apoptotic cells was distinct. Fibroblasts from granulation tissue and skin explants were placed in a collagen gel contraction assay prior to the 5th passage to determine their in vitro contractility. While granulation tissue fibroblasts from young and old rats showed similar collagen gel contractility, skin fibroblasts from old rats displayed greater collagen gel contractile behavior than young skin fibroblasts. Greater gel contractility of fibroblasts from old rats appeared to result, in large part, from the ability of those cells to cause generalized gel degradation. Gelatin zymography indicated a greater abundance of matrix metalloproteinase-2 in supernatants from gels containing skin fibroblasts from old rats. Taken together, these results suggest that the age-associated healing delay in the rat may not be related to the appearance or abundance of distinct myofibroblast or apoptotic cell populations. Proteolysis may have a significant role in delayed wound healing in aged animals. PMID- 11472620 TI - Development of angiotensin (1-7) as an agent to accelerate dermal repair. AB - Angiotensin II has been shown to be a potent agent in the acceleration of wound repair. Angiotensin (1-7), a fragment of angiotensin II that is not hypertensive, was found to be comparable to angiotensin II in accelerating dermal healing. This activity was evaluated in four models: rat and diabetic mouse full-thickness excisional wounds; rat random flap; and guinea pig partial thickness thermal injury. In all models, angiotensin (1-7) was comparable to angiotensin II. Angiotensin (1-7) accelerated the closure of wounds in diabetic mice and rats. In diabetic mice the resultant tissue at day 25 after injury was more comparable to normal tissue than the fibrotic scar observed in placebo-treated wounds. In the random flap model, angiotensin (1-7) was comparable to angiotensin II in maintaining flap viability (approximately 82%) and flap survival (40%). Finally, angiotensin (1-7) increased proliferation in the hair follicles at the edge of the wound and site of thermal injury, and the number of patent blood vessels on day 7 after partial thickness thermal injury. These data may be partially explained by the effect of angiotensin II and angiotensin (1-7) on keratinocyte proliferation. While platelet-derived growth factor had no effect on keratinocyte proliferation, angiotensin II and angiotensin (1-7) significantly increased keratinocyte proliferation. These data show that angiotensin(1-7) is comparable to angiotensin II in accelerating skin repair. Furthermore, the hypertensive and wound healing effects can be separated within the family of angiotensin peptides. PMID- 11472621 TI - Laser photostimulation accelerates wound healing in diabetic rats. AB - In this study, we examined the hypothesis that laser photostimulation can facilitate healing of impaired wounds in experimental diabetes using a rat model. Diabetes was induced in male rats by streptozotocin injection and two 6 mm diameter circular wounds were created on either side of the spine. The left wound of each animal was treated with a 632.8 nm He:Ne laser at a dose of 1.0 J/cm2 for five days a week until the wounds closed (three weeks). Measurements of the biomechanical properties of the laser-treated wounds indicated there was a marginal increase in maximum load (16%), stress (16%), strain (27%), energy absorption (47%) and toughness (84%) compared to control wounds of diabetic rats. Biochemical assays revealed that the amount of total collagen was significantly increased in laser treated wounds (274 +/- 8.7 microg) over the control wounds (230 +/- 8.4 microg). Sequential extractions of collagen from healing wounds showed that laser treated wounds had significantly greater concentrations of neutral salt soluble (15%) and insoluble collagen (16%) than control wounds, suggesting accelerated collagen production in laser treated wounds. There was an appreciable decrease in pepsin soluble collagen (19%) in laser treated wounds over control wounds, indicating higher resistance to proteolytic digestion. In conclusion, the biomechanical and biochemical results collectively suggest that laser photostimulation promotes the tissue repair process by accelerating collagen production and promoting overall connective tissue stability in healing wounds of diabetic rats. PMID- 11472622 TI - New FDA xenotransplantation documents: a proposed rule and a draft guidance. PMID- 11472623 TI - Effects of specific anti-B and/or anti-plasma cell immunotherapy on antibody production in baboons: depletion of CD20- and CD22-positive B cells does not result in significantly decreased production of anti-alphaGal antibody. AB - Anti-Galalpha1-3Gal antibodies (antialphaGal Ab) are a major barrier to clinical xenotransplantation as they are believed to initiate both hyperacute and acute humoral rejection. Extracorporeal immunoadsorption (EIA) with alphaGal oligosaccharide columns temporarily depletes antialphaGal Ab, but their return is ultimately associated with graft destruction. We therefore assessed the ability of two immunotoxins (IT) and two monoclonal antibodies (mAb) to deplete B and/or plasma cells both in vitro and in vivo in baboons, and to observe the rate of return of antialphaGal Ab following EIA. The effects of the mouse anti-human IT anti-CD22-ricin A (proportional to CD22-IT, directed against a B cell determinant) and anti-CD38-ricin A (proportional to CD38-IT, B and plasma cell determinant) and the mouse anti-human anti-CD38 mAb (proportional to CD38 mAb) and mouse/human chimeric anti-human anti-CD20 mAb (proportional to CD20 mAb, Rituximab, B cell determinant) on B and plasma cell depletion and antialphaGal Ab production were assessed both in vitro and in vivo in baboons (n = 9) that had previously undergone splenectomy. For comparison, two baboons received nonmyeloablative whole body irradiation (WBI) (300 cGy), and one received myeloablative WBI (900 cGy). Depletion of B cells was monitored by flow cytometry of blood, bone marrow (BM) and lymph nodes (LN), staining with anti-CD20 and/or anti-CD22 mAbs, and by histology of LN. EIA was carried out after the therapy and antialphaGal Ab levels were measured daily. In vitro proportional to CD22-IT inhibited protein synthesis in the human Daudi B cell line more effectively than proportional to CD38-IT. Upon differentiation of B cells into plasma cells, however, less inhibition of protein synthesis after proportional to CD22-IT treatment was observed. Depleting CD20-positive cells in vitro from a baboon spleen cell population already depleted of granulocytes, monocytes, and T cells led to a relative enrichment of CD20-negative cells, that is plasma cells, and consequently resulted in a significant increase in antialphaGal Ab production by the remaining cells, whereas depleting CD38-positive cells resulted in a significant decrease in antialphaGal Ab production. In vivo, WBI (300 or 900 cGy) resulted in 100% B cell depletion in blood and BM, > 80% depletion in LN, with substantial recovery of B cells after 21 days and only transient reduction in antialphaGal Ab after EIA. Proportional to CD22-IT depleted B cells by > 97% in blood and BM, and by 60% in LN, but a rebound of B cells was observed after 14 and 62 days in LN and blood, respectively. At 7 days, serum antialphaGal IgG and IgM Ab levels were reduced by a maximum of 40-45% followed by a rebound to levels up to 12-fold that of baseline antialphaGal Ab by day 83 in one baboon. The results obtained with proportional to CD38-IT were inconclusive. This may have been, in part, due to inadequate conjugation of the toxin. Cell coating was 100% with proportional to CD38 mAb, but no changes in antialphaGal Ab production were observed. Proportional to CD20 mAb resulted in 100% depletion of B cells in blood and BM, and 80% in LN, with recovery of B cells starting at day 42. Adding 150cGy WBI at this time led to 100% depletion of B cells in the BM and LN. Although B cell depletion in blood and BM persisted for > 3 months, the reduction of serum antialphaGal IgG or IgM Ab levels was not sustained beyond 2 days. Proportional to CD20 mAb + WBI totally and efficiently depleted CD20- and CD22-positive B cells in blood, BM, and LN for > 3 months in vivo, but there was no sustained clinically significant reduction in serum antialphaGal Ab. The majority of antibody secretors are CD38-positive cells, but targeting these cells in vitro or in vivo with proportional to CD38-IT was not very effective. These observations suggest that CD20-and CD22-positive B cells are not the major source of antialphaGal Ab production. Future efforts will be directed towards suppression of plasma cell function. PMID- 11472624 TI - Detection of xenoantibodies using a simple flow cytometric assay. AB - Higher primates, including humans, have high levels of pre-existing naturally circulating antibodies that predominantly recognize the epitope Gal (1,3-Gal), which is highly expressed on the surface of xenogenic cells. Deposition of these antibodies on the endothelial cell surface of vascularized xenografts leads to an activation of the classical pathway of the complement system, resulting in tissue ischemia and necrosis with rapid demise of the xenograft. This hyperacute rejection (HAR) is always a major barrier in xenograft transplantation and should be minimized by accurately monitoring the naturally occurring antibodies. In the present study, we utilized a simple and rapid flow cytometric (FCM) assay to monitor the presence of these naturally occurring antibodies. We found that the FCM assay is very effective in measuring human antibodies bound to the xenogenic cells, which cause cytotoxicity. This assay could be useful in the pre- and post xenotransplantation monitoring of xenoantibodies, thus, helping in the development of strategies to block the binding of preformed human antibodies to the xenograft in order to overcome the problem of HAR. PMID- 11472625 TI - A new in vitro model for the study of pig-to-human vascular hyperacute rejection. AB - Studies on vascular hyperacute xenograft rejection (HAR) are usually conducted in vitro on cultured endothelial cells (EC) exposed to human serum, in complex whole organ perfusion models using heparinized blood or in vivo models. Here we describe a new model allowing perfusion of pig vessels with human whole blood without anticoagulants. Segments of the porcine iliac artery were connected to circular polyvinyl chloride (PVC) tubing, whose inner surface was conjugated with immobilized heparin. The vessels were perfused with 7 to 8 ml of fresh, non anticoagulated human blood by rocking of the tubing device for 5, 15 or 60 min in an incubator at 37 degrees C. Human iliac arteries (n = 4) were perfused with fresh human ABO-compatible blood as controls. Perfusion of human vessels resulted in changes in the blood and plasma parameters similar to those in the PVC control loop. Overall, perfusion of the porcine vessels generated high levels of C3a, sC5b-9 and thrombin-anti-thrombin (TAT). Platelet consumption was near total (97.2 +/- 1.2%; "high" responders) in six of 13 vessels perfused and only moderate (55.8 +/- 9.9%; "low" responders) in the remaining seven vessels. The "high" responder vessel group showed a significantly higher platelet reduction, neutrophil loss and monocyte consumption and higher C3a and TAT factor at 60 min compared with the human vessels. The "low" responder porcine vessel group also generated significantly higher TAT levels at 60 min compared with the human vessels, but lower levels compared with the "high" responder porcine vessel group. Immunohistochemical examination of perfused porcine vessels revealed binding of human IgM, IgG, IgA, C1q, C3, fibrin and platelets at 5 min. The binding of these proteins was even stronger at 15 and 60 min, and at 60 min C9 could also be detected. Addition of soluble complement receptor 1 (sCR1) to the blood resulted in a significant reduction in C3a and sC5b-9 (P = 0.046 and P = 0.046, respectively). However, sCR1 did not reduce C1q, C3c or C5 staining, but did abolish C9 binding to the endothelium. In conclusion, in vitro perfusion of porcine vessel segments with non-heparinized, fresh human blood triggered events characterizing HAR. The small quantity of blood and xenogenic tissue that is needed makes this model ideal for investigations of the mechanisms and treatments of rejections associated with xenogeneic pig-to-human xenotransplantation. PMID- 11472626 TI - T-cell differentiation of human and non-human primate CD34+ hematopoietic progenitor cells using porcine thymic stroma. AB - Transplantation of swine thymic tissue has been proposed as an approach to reconstitute the immune system of HIV-infected individuals. This is an attractive strategy because miniature swine are readily available as donors and porcine tissue is resistant to infection with HIV-1. Demonstration that porcine thymus tissue supports primate T-cell differentiation is critical to the ultimate utility of this approach. Using a thymic stroma culture system we have previously described [Rosenzweig M, Marks DF, Zhu H et al. In vitro T lymphopoiesis of human and rhesus CD34+ progenitor cells. Blood 1996; 87: 4040], we demonstrate that porcine thymus tissue is able to promote the in vitro T-lymphocyte differentiation of both human and non-human primate hematopoietic progenitor cells. CD34+ hematopoietic progenitors differentiated into both double positive (CD4+CD8+) and single positive thymocytes expressing CD4 or CD8 alone. A polyclonal T-cell repertoire was evident. In addition, the T cells responded appropriately to mitogen and were permissive to infection with simian immunodeficiency virus (SIV). These data demonstrate the ability of porcine thymus to support T-cell differentiation of both human and non-human hematopoietic progenitor cells and support in vivo studies of transplantation of swine thymic tissue as a strategy for immune reconstitution in AIDS. PMID- 11472627 TI - Effect in vitro and in vivo of a rat anti-CD2 monoclonal antibody (LO-CD2b) on pig-to-baboon xenogeneic cellular (T and natural killer cells) immune response. AB - Although hyperacute rejection of discordant xenogeneic grafts can be prevented, baboon or human anti-pig cellular response may lead to acute xenograft rejection. Among the immune cellular actors participating in such a xenograft rejection are both T and natural killer (NK) cells. In the pre-clinical model of pig-to-baboon discordant xenograft, there is however, a lack of specific immunological therapeutic agent, in particular antibaboon T-cell monoclonal antibodies do not exist. We therefore developed a rat anti-CD2 monoclonal antibody (LO-CD2b) that recognizes both baboon and human CD2 + cells. In this study, we show that in vitro LO-CD2b inhibits a pig-to-baboon mixed lymphocyte reaction, the direct cytotoxicity of baboon peripheral blood lymphocytes to pig aortic endothelial cells, as well as the baboon NK activity against K562 cell line. In vivo, LO-CD2b produces a strong depletion of all peripheral CD2+ cells including NK CD2+ cells. In summary, LO-CD2b represents an important immunological tool that can be used in the preclinical model of discordant pig-to-baboon vascularized xenograft. PMID- 11472628 TI - The in vitro activity and specificity of human endothelial cell-specific promoters in porcine cells. AB - The chronic shortage of human organs, tissues and cells for transplantation has inspired research on the possibility of using animal donor tissue instead. Transplantation over a species barrier is associated with rejections which are difficult to control. Therefore, it is generally agreed that successful pig to human xenotransplantation requires donor pigs to be genetically modified. Vascular endothelium is the most immediate barrier between the xenogeneic donor organ and host immune and nonimmune defense systems. Thus, these cells are the prime targets for such genetic modifications. Luciferase assays were used to evaluate the activity and specificity of human endothelial-cell specific promoters in porcine aortic-, microvascular- and nonendothelial cells. The promoters for human Flk-1 (fetal liver kinase-1), Flt-1 (fms-like tyrosine kinase), ICAM-2 (intercellular adhesion molecule-2), thrombomodulin and vWf (von Willebrand factor) supported similar levels of luciferase expression in human and porcine aortic endothelial cells, with the Flk-1 promoter being the strongest followed by the thrombomodulin promoter. Relative to the activity of the CMV promoter, the human endothelial cell-specific promoters all showed less activity in porcine kidney microvascular endothelial cells than in liver or brain microvascular endothelial cells. The thrombomodulin and Flk-1 promoters exhibited similar activity in liver and kidney microvascular endothelial cells, whereas the Flk-1 promoter was stronger in aortic and brain microvascular endothelial cells. Human endothelial cell-specific promoters also showed some degree of specificity in pig, because they supported less luciferase activity in porcine nonendothelial cell lines. Based on the in vitro data and previously published in vivo data, the human Flk-1 and thrombomodulin promoters are good candidate promoters for strong endothelial cell-specific gene expression in transgenic pigs. PMID- 11472629 TI - Hamster-to-rat bone marrow xenotransplantation and humoral graft vs. host disease. AB - Bone marrow transplantation (BMT) may induce tolerance across xenogeneic barriers. We have established a xenogeneic BMT model where hamster BM is transplanted into splenectomized LEW rat recipients resulting in high levels of engraftment. Unfortunately, graft vs. host disease (GVHD) with severe dermatitis developed in all rat recipients. We were successful in treating or preventing the dermatitis of this xenogeneic GVHD by the use of the T-cell suppressant tacrolimus. However, this compound did not prevent the development of a fatal liver injury in the rat recipients. This study was designed to elucidate the pathogenesis of this liver injury appearing in T-cell suppressed rat recipients of hamster BM. Splenectomized and irradiated (10 Gy) LEW rats received 300 x 106 unfractionated hamster BM cells. These BMT recipients were divided in 3 groups: Group I recipients (n = 8) did not receive further immunosuppression. Group II animals (n = 10) received tacrolimus 1 mg/kg/d for 7 d. Group III recipients (n = 6) were given the same daily dose of tacrolimus on a long-term basis. Chimerism was detected by flow cytometry. Cytotoxicity of recipient's sera against rat and hamster lymph node cells was measured by complement-dependent cytotoxicity (CDC) test. Immunofluorescence was used to detect hamster antirat antibodies on several recipient organs. In Group I, 2 out of 8 animals engrafted (25%) and survived for a median of 21 d showing the severe dermatitis characteristic of GVHD. In group II (n = 10), 9/10 rat recipients engrafted (90%) and survival was increased to a median of 53.7 days. However, these surviving recipients developed fatal GVHD not different from that observed in Group I recipients. All animals in Group III (n = 6) engrafted and did not show the characteristic dermatitis of GVHD. Their survival, however, was shortened to a median of 30.3 d by a severe liver injury. This injury was characterized by hepatocyte necrosis in zones 1 and 2 with polymorphonuclear (PMN) cell infiltration. Deposits of hamster immunoglobulins were present around the necrotic areas and in the portal veins. Moreover, antirat antibodies appeared in the circulation. These antibodies were sensitive to dithiothreitol (DTT) treatment indicating that they were of the IgM class. This study shows that xenogeneic GVHD may have a dual presentation in the hamster-to rat model: a classical cellular GVHD not distinct to the allogeneic one and a humoral GVHD affecting solely the recipient liver. The degree of humoral injury is potentiated by T-cell suppression. PMID- 11472630 TI - mRNA:guanine-N7 cap methyltransferases: identification of novel members of the family, evolutionary analysis, homology modeling, and analysis of sequence structure-function relationships. AB - BACKGROUND: The 5'-terminal cap structure plays an important role in many aspects of mRNA metabolism. Capping enzymes encoded by viruses and pathogenic fungi are attractive targets for specific inhibitors. There is a large body of experimental data on viral and cellular methyltransferases (MTases) that carry out guanine-N7 (cap 0) methylation, including results of extensive mutagenesis. However, a crystal structure is not available and cap 0 MTases are too diverged from other MTases of known structure to allow straightforward homology-based interpretation of these data. RESULTS: We report a 3D model of cap 0 MTase, developed using sequence-to-structure threading and comparative modeling based on coordinates of the glycine N-methyltransferase. Analysis of the predicted structural features in the phylogenetic context of the cap 0 MTase family allows us to rationalize most of the experimental data available and to propose potential binding sites. We identified a case of correlated mutations in the cofactor-binding site of viral MTases that may be important for the rational drug design. Furthermore, database searches and phylogenetic analysis revealed a novel subfamily of hypothetical MTases from plants, distinct from "orthodox" cap 0 MTases. CONCLUSIONS: Computational methods were used to infer the evolutionary relationships and predict the structure of Eukaryotic cap MTase. Identification of novel cap MTase homologs suggests candidates for cloning and biochemical characterization, while the structural model will be useful in designing new experiments to better understand the molecular function of cap MTases. PMID- 11472631 TI - The C-terminal domain of the Bloom syndrome DNA helicase is essential for genomic stability. AB - BACKGROUND: Bloom syndrome is a rare cancer-prone disorder in which the cells of affected persons have a high frequency of somatic mutation and genomic instability. Bloom syndrome cells have a distinctive high frequency of sister chromatid exchange and quadriradial formation. BLM, the protein altered in BS, is a member of the RecQ DNA helicase family, whose members share an average of 40% identity in the helicase domain and have divergent N-terminal and C-terminal flanking regions of variable lengths. The BLM DNA helicase has been shown to localize to the ND10 (nuclear domain 10) or PML (promyelocytic leukemia) nuclear bodies, where it associates with TOPIIIalpha, and to the nucleolus. RESULTS: This report demonstrates that the N-terminal domain of BLM is responsible for localization of the protein to the nuclear bodies, while the C-terminal domain directs the protein to the nucleolus. Deletions of the N-terminal domain of BLM have little effect on sister chromatid exchange frequency and chromosome stability as compared to helicase and C-terminal mutations which can increase SCE frequency and chromosome abnormalities. CONCLUSION: The helicase activity and the C-terminal domain of BLM are critical for maintaining genomic stability as measured by the sister chromatid exchange assay. The localization of BLM into the nucleolus by the C-terminal domain appears to be more important to genomic stability than localization in the nuclear bodies. PMID- 11472632 TI - Kinesins in the Arabidopsis genome: a comparative analysis among eukaryotes. AB - BACKGROUND: Kinesins constitute a superfamily of microtubule motor proteins that are found in eukaryotic organisms. Members of the kinesin superfamily perform many diverse cellular functions such as transport of vesicles and organelles, spindle formation and elongation, chromosome segregation, microtubule dynamics and morphogenesis. Only a few kinesins have been characterized in plants including Arabidopsis thaliana. Because of the diverse cellular functions in which kinesins are involved, the number, types and characteristics of kinesins present in the Arabidopsis genome would provide valuable information for many researchers. RESULTS: Here we have analyzed the recently completed Arabidopsis genome sequence and identified sixty-one kinesin genes in the Arabidopsis genome. Among the five completed eukaryotic genomes the Arabidopsis genome has the highest percentage of kinesin genes. Further analyses of the kinesin gene products have resulted in identification of several interesting domains in Arabidopsis kinesins that provide clues in understanding their functions. A phylogenetic analysis of all Arabidopsis kinesin motor domain sequences with 113 motor domain sequences from other organisms has revealed that Arabidopsis has seven of the nine recognized subfamilies of kinesins whereas some kinesins do not fall into any known family. CONCLUSION: There are groups of Arabidopsis kinesins that are not present in yeast, Caenorhabditis elegans and Drosophila melanogaster that may, therefore, represent new subfamilies specific to plants. The domains present in different kinesins may provide clues about their functions in cellular processes. The comparative analysis presented here provides a framework for future functional studies with Arabidopsis kinesins. PMID- 11472633 TI - Identification of the gene encoding Brain Cell Membrane Protein 1 (BCMP1), a putative four-transmembrane protein distantly related to the Peripheral Myelin Protein 22 / Epithelial Membrane Proteins and the Claudins. AB - BACKGROUND: A partial cDNA clone from dog thyroid presenting a very significant similarity with an uncharacterized mouse EST sequence was isolated fortuitously. We report here the identification of the complete mRNA and of the gene, the product of which was termed "brain cell membrane protein 1" (BCMP1). RESULTS: The 4 kb-long mRNA sequence exhibited an open-reading frame of only 543 b followed by a 3.2 kb-long 3' untranslated region containing several AUUUA instability motifs. Analysis of the encoded protein sequence identified the presence of four putative transmembrane domains. Similarity searches in protein domain databases identified partial sequence conservations with peripheral myelin protein 22 (PMP22)/ epithelial membrane proteins (EMPs) and Claudins, defining the encoded protein as representative of the existence of a novel subclass in this protein family.Northern-blot analysis of the expression of the corresponding mRNA in adult dog tissues revealed the presence of a huge amount of the 4 kb transcript in the brain. An EGFP-BCMP1 fusion protein expressed in transfected COS-7 cells exhibited a membranous localization as expected. The sequences encoding BCMP1 were assigned to chromosome X in dog, man and rat using radiation hybrid panels and were partly localized in the currently available human genome sequence. CONCLUSIONS: We have identified the existence in several mammalian species of a gene encoding a putative four-transmembrane protein, BCMP1, wich defines a novel subclass in this family of proteins. In dog at least, the corresponding mRNA is highly present in brain cells. The chromosomal localization of the gene in man makes of it a likely candidate gene for X-linked mental retardation. PMID- 11472634 TI - Low frequency of mutations in the core promoter and precore regions of hepatitis B virus in anti-HBe positive Brazilian carriers. AB - BACKGROUND: Mutations in the core promoter and precore regions of the hepatitis B virus (HBV) genome, notably the double substitution (AGG to TGA) at nt positions 1762-1764 in the core promoter, and the precore stop codon mutation G to A at nt 1896, can often explain the anti-HBe phenotype in chronic carriers. However, the A1896 mutation is restricted to HBV isolates that have T at nt 1858. The double substitution at positions 1762-1764 has been described to occur preferentially in patients infected with strains showing C instead of T at nt 1858. RESULTS: HBV DNAs from 29 anti-HBe Brazilian samples were characterized by nucleotide sequencing of PCR products from precore region. Among them, 18 isolates presented C at nt 1858 (mostly genotype A strains). The 11 remaining isolates (genotypes D and F) had T1858. The stop codon mutation at nt 1896 was found in seven isolates (24% of the total and 63% of the isolates that had T1858). The frequency of the double substitution at positions 1762-1764 was surprisingly low (20%) among C1858 isolates. An association between A1896 and TGA 1762-1764 mutations was observed among genotype D isolates: these showed either none of the two mutations or both. Furthermore, strains mutated at positions 1896 and/or 1762-1764 also presented an elevated number of other, less common substitutions in the core promoter and precore regions. CONCLUSIONS: The data reported here are not in accordance with some reports from other parts of the world. In half of the isolates, none of the mutations previously described could explain the anti-HBe phenotype. PMID- 11472635 TI - The estimated economic burden of genital herpes in the United States. An analysis using two costing approaches. AB - BACKGROUND: Only limited data exist on the costs of genital herpes (GH) in the USA. We estimated the economic burden of GH in the USA using two different costing approaches. METHODS: The first approach was a cross-sectional survey of a sample of primary and secondary care physicians, analyzing health care resource utilization. The second approach was based on the analysis of a large administrative claims data set. Both approaches were used to generate the number of patients with symptomatic GH seeking medical treatment, the average medical expenditures and estimated national costs. Costs were valued from a societal and a third party payer's perspective in 1996 US dollars. RESULTS: In the cross sectional study, based on an estimated 3.1 million symptomatic episodes per year in the USA, the annual direct medical costs were estimated at a maximum of $984 million. Of these costs, 49.7% were caused by drug expenditures, 47.7% by outpatient medical care and 2.6% by hospital costs. Indirect costs accounted for further $214 million. The analysis of 1,565 GH cases from the claims database yielded a minimum national estimate of $283 million direct medical costs. CONCLUSIONS: GH appears to be an important public health problem from the health economic point of view. The observed difference in direct medical costs may be explained with the influence of compliance to treatment and possible undersampling of subpopulations in the claims data set. The present study demonstrates the validity of using different approaches in estimating the economic burden of a specific disease to the health care system. PMID- 11472636 TI - Clinical features of culture-proven Mycoplasma pneumoniae infections at King Abdulaziz University Hospital, Jeddah, Saudi Arabia. AB - OBJECTIVE: This retrospective chart review describes the epidemiology and clinical features of 40 patients with culture-proven Mycoplasma pneumoniae infections at King Abdulaziz University Hospital, Jeddah, Saudi Arabia. METHODS: Patients with positive M. pneumoniae cultures from respiratory specimens from January 1997 through December 1998 were identified through the Microbiology records. Charts of patients were reviewed. RESULTS: 40 patients were identified, 33 (82.5%) of whom required admission. Most infections (92.5%) were community acquired. The infection affected all age groups but was most common in infants (32.5%) and pre-school children (22.5%). It occurred year-round but was most common in the fall (35%) and spring (30%). More than three-quarters of patients (77.5%) had comorbidities. Twenty-four isolates (60%) were associated with pneumonia, 14 (35%) with upper respiratory tract infections, and 2 (5%) with bronchiolitis. Cough (82.5%), fever (75%), and malaise (58.8%) were the most common symptoms, and crepitations (60%), and wheezes (40%) were the most common signs. Most patients with pneumonia had crepitations (79.2%) but only 25% had bronchial breathing. Immunocompromised patients were more likely than non immunocompromised patients to present with pneumonia (8/9 versus 16/31, P = 0.05). Of the 24 patients with pneumonia, 14 (58.3%) had uneventful recovery, 4 (16.7%) recovered following some complications, 3 (12.5%) died because of M pneumoniae infection, and 3 (12.5%) died due to underlying comorbidities. The 3 patients who died of M pneumoniae pneumonia had other comorbidities. CONCLUSION: our results were similar to published data except for the finding that infections were more common in infants and preschool children and that the mortality rate of pneumonia in patients with comorbidities was high. PMID- 11472637 TI - MCM2--a promising marker for premalignant lesions of the lung: a cohort study. AB - BACKGROUND: Because cells progressing to cancer must proliferate, marker proteins specific to proliferating cells may permit detection of premalignant lesions. Here we compared the sensitivities of a classic proliferation marker, Ki-67, with a new proliferation marker, MCM2, in 41 bronchial biopsy specimens representing normal mucosa, metaplasia, dysplasia, and carcinoma in situ. METHODS: Parallel sections were stained with antibodies against MCM2 and Ki-67, and the frequencies of staining were independently measured by two investigators. Differences were evaluated statistically using the two-sided correlated samples t-test and Wilcoxon rank sum test. RESULTS: For each of the 41 specimens, the average frequency of staining by anti-MCM2 (39%) was significantly (p < 0.001) greater than by anti-Ki-67 (16%). In metaplastic lesions anti-MCM2 frequently detected cells near the epithelial surface, while anti-Ki-67 did not. CONCLUSIONS: We conclude that MCM2 is detectable in 2-3 times more proliferating premalignant lung cells than is Ki-67. The promise of MCM2 as a sensitive marker for premalignant lung cells is enhanced by the fact that it is present in cells at the surface of metaplastic lung lesions, which are more likely to be exfoliated into sputum. Future studies will determine if use of anti-MCM2 makes possible sufficiently early detection to significantly enhance lung cancer survival rates. PMID- 11472638 TI - Herpes simplex virus bullous keratitis misdiagnosed as a case of pseudophakic bullous keratopathy with secondary glaucoma: an unusual presentation. AB - PURPOSE: To report an unusual case of herpetic bullous keratitis misdiagnosed as a case of pseudophakic bullous keratopathy with secondary glaucoma. RESULTS: A retrospective analysis of the case record of a 60-year-old man who had earlier undergone bilateral cataract surgery, was done. He presented with a complaint of decrease in vision in the right eye of 20 days duration. On examination, cornea showed epithelial bullae all over the surface with stromal and epithelial edema. Intraocular pressure was 30 mm of Hg in RE. He was treated with anti-glaucoma medications. Two dendritic lesions were seen in the cornea during a subsequent visit four days later. Virological investigations confirmed a diagnosis of Herpes simplex keratitis. He was treated with topical acyclovir. CONCLUSIONS: This case highlights the fact that herpes simplex keratitis can present initially as a more diffuse corneal stromal and epithelial edema with epithelial bullae mimicking bullous keratopathy. Herpetic bullous keratitis, although unusual, should be considered in the differential diagnosis under such circumstances. PMID- 11472639 TI - A geometric and algebraic view of MHC-peptide complexes and their binding properties. AB - BACKGROUND: Major histocompatibility complex (MHC) molecules present peptides to T lymphocytes. It is of critical biological and medical importance to elucidate how different MHC alleles bind to a specific set of peptides. METHOD: In this study we approach the problem from the algebraic and geometric point of view to analyse MHC-peptide-binding data accumulated over the years. The space of sequence properties (having a particular amino acid at a particular position) of MHC-peptide complexes conveys a geometric structure to these sequence properties in the form of a distance measure, which reveals the peptide binding requirements imposed by the polymorphic sequence characteristics of the MHC molecules. RESULTS: Comparison of the results of this study with our current knowledge of MHC-peptide binding constraints leads to robust agreement. This study provides the tools to quantitate these binding constraints giving a more detailed account of them and opening the way to make peptide binding predictions for MHC alleles for which there is no peptide elution data. In addition, the geometric representation of MHC-peptide complex sequence data gives a distance measure between amino acids in reference to their ability to meet MHC binding requirements. CONCLUSIONS: The algebraic and geometric view of amino acid sequences provides a theoretical framework to study the function of proteins when there is enough variation in this sequence to account for the variation in their function, as it is the case with MHC molecules in regard to their ability to present peptides. PMID- 11472640 TI - Appendicectomies in Albanians in Greece: outcomes in a highly mobile immigrant patient population. AB - BACKGROUND: Albanian immigrants in Greece comprise a highly mobile population with unknown health care profile. We aimed to assess whether these immigrants were more or less likely to undergo laparotomy for suspected appendicitis with negative findings (negative appendicectomy), by performing a controlled study with individual (1:4) matching. We used data from 6 hospitals in the Greek prefecture of Epirus that is bordering Albania. RESULTS: Among a total of 2027 non-incidental appendicectomies for suspected appendicitis performed in 1994 1999, 30 patients with Albanian names were matched (for age, sex, time of operation and hospital) to 120 patients with Greek names. The odds for a negative appendicectomy were 3.4-fold higher (95% confidence interval [CI], 1.24-9.31, p = 0.02) in Albanian immigrants than in matched Greek-name subjects. The difference was most prominent in men (odds ratio 20.0, 95% CI, 1.41-285, p = 0.02) while it was not formally significant in women (odds ratio 1.56, 95% CI, 0.44-5.48). The odds for perforation were 1.25-fold higher in Albanian-name immigrants than in Greek-name patients (95% CI 0.44- 3.57). CONCLUSIONS: Albanian immigrants in Greece are at high risk for negative appendicectomies. Socioeconomic, cultural and language parameters underlying health care inequalities in highly mobile immigrant populations need better study. PMID- 11472641 TI - A decision analytical cost analysis of offering ECV in a UK district general hospital. AB - OBJECTIVE: To determine the care pathways and implications of offering mothers the choice of external cephalic version (ECV) at term for singleton babies who present with an uncomplicated breech pregnancy versus assisted breech delivery or elective caesarean. DESIGN: A prospective observational audit to construct a decision analysis of uncomplicated full term breech presentations. SETTING: The North Staffordshire NHS Trust. SUBJECTS: All women (n = 176) who presented at full term with a breech baby without complications during July 1995 and June 1997. MAIN OUTCOME MEASURES: The study determined to compare the outcome in terms of the costs and cost consequences for the care pathways that resulted from whether a women chose to accept the offer of ECV or not. All the associated events were then mapped for the two possible pathways. The costs were considered only within the hospital setting, from the perspective of the health care provider up to the point of delivery. RESULTS: The additional costs for ECV, assisted breech delivery and elective caesarean over and above a normal birth were 186.70 pounds sterling, 425.36 pounds sterling and 1,955.22 pounds sterling respectively. The total expected cost of the respective care pathways for "ECV accepted" and "ECV not accepted" (including the probability of adverse events) were 1,452 pounds sterling and 1,828 pounds sterling respectively, that is the cost of delivery through the ECV care pathways is less costly than the non ECV delivery care pathway. CONCLUSIONS: Implementing an ECV service may yield cost savings in secondary care over and above the traditional delivery methods for breech birth of assisted delivery or caesarean section. The scale of these expected cost savings are in the range of 248 pounds sterling to 376 pounds sterling per patient. This converts to a total expected cost saving of between 43,616 pounds sterling and 44,544 pounds sterling for the patient cohort considered in this study. PMID- 11472648 TI - Undifferentiated connective tissue diseases: a new frontier in rheumatology. PMID- 11472652 TI - Editorial. AB - PMID- 11472651 TI - Editorial: Dedicated to the Memory of Marshall R. Urist. AB - BACKGROUND: Previous experimental studies have shown a higher degree of bone implant contact for surface-enlarged implants compared with machined implants. Yet, there is insufficient evidence that such implants show higher stability and an increased survival rate. PURPOSE: The purpose of this investigation was to study the integration and stability of grit-blasted implants with retention elements on the implant neck, with and without marginal bone defects, compared with machined implants without retention elements. MATERIALS AND METHODS: After tooth extraction of the mandibular premolars in six dogs, two grit-blasted, partly microthreaded Astra Tech implants and one standard Branemark implant were bilaterally placed in each dog. On one side, 3 yen 3 mm large buccal defects were created, to expose three to four implant threads. The contralateral side served as control, and no defects were made. The animals were sacrificed after 4 months of healing. Implant stability was measured using resonance frequency analysis at implant installation and after 4 months of healing. Histologic and histomorphometric evaluation was made after 4 months of healing. RESULTS: Resonance frequency analysis indicated that all implants in the test and control groups were osseointegrated after 4 months, with a tendency toward higher implant stability for the Astra Tech implants. There was a statistically significant higher increase in resonance frequency for the Astra test implants compared with their corresponding controls. Histology and histomorphometry showed well integrated implants with varying degrees of bone repair at the defect sites. The greater bone-implant contact for the Astra implants was statistically significant. No significant difference between the implants in amount of bone filling the threads was recorded. CONCLUSIONS: The Astra Tech implants tested showed a higher degree of bone-implant contact and higher level of bone regenerated at defect sites compared with the Branemark implants. Resonance frequency analysis demonstrated a significantly higher increase in the Astra test implants compared with their control groups than did the Branemark test implants versus their controls. PMID- 11472653 TI - In search of truth: the role of systematic reviews and meta-analyses for assessing the effectiveness of rehabilitation with oral implants. AB - BACKGROUND: It is difficult to determine the effectiveness or potential harm of dental therapies. Thus, any tools able to condense reliable scientific information would be of benefit. PURPOSE: To discuss methods for the assessment of the scientific literature and, in particular, of systematic reviews for evaluating the effectiveness of oral implant rehabilitation procedures. MATERIALS AND METHODS: Various study designs and methods to identify scientific evidence are described, discussed, and ranked. Issues on how to critically appraise randomized controlled clinical trials (RCTs) and systematic reviews are presented. RESULTS: Properly conducted RCTs and structured critical systematic reviews are the gold standard of clinical research for assessing whether a therapeutic intervention is effective. In the field of oral implantology, there is an urgent need to implement more RCTs and to summarize their results in systematic reviews. CONCLUSIONS: Searching of the truth requires training and critical skill. Dentists should be trained on how to integrate their clinical experience with evidence-based research and on how to discriminate between clinically useful scientific information and less useful research. PMID- 11472654 TI - Immediate provisional for single-tooth implant replacement with Branemark system: preliminary report. AB - BACKGROUND: The success of osseointegrated implants ad modum Branemark for single tooth restorations is documented. Future developments should aim at improving the benefits to patients by decreasing treatment time, minimizing surgical stages, and maximizing esthetic outcomes. Using knowledge from studies of immediate implant placement, one-stage, immediate loading protocols, the authors developed the immediate provisional. PURPOSE: The purpose of this study was to develop a protocol to provide an immediate solution for restoring a single missing tooth in the esthetic zone. The protocol should be simple, predictable, cost effective, and allow the use of other techniques to improve esthetic outcome. MATERIALS AND METHOD: This prospective clinical study included 24 patients treated from August 1999 to October 2000. Single-tooth implant replacement was done according to immediate provisional protocol. Thirteen of the 24 patients had immediate implant placement after tooth extraction. All implants were placed in the esthetic zone. During surgery, emphasis was placed on obtaining primary stability by achieving bicortical anchorage and maximum insertion torque of at least 40 Ncm. CeraOne (Nobel Biocare) abutments were used, and provisional crowns were fabricated immediately before wound closure. The occlusion was protected by adjacent teeth. RESULTS: Within the follow-up period of between 1 month and 15 months, all fixtures in the 24 patients were stable. Crestal bone loss greater than one thread-width was not detected. The esthetic result was considered satisfactory by all patients. CONCLUSIONS: The implant placement and restoration protocol used in this study showed promising initial results for both the immediate implant and healed extraction site groups. The desirable goals of patient satisfaction, excellent esthetic outcomes, and no increase in treatment cost were achieved in this protocol. Further studies to elucidate the potential of the immediate provisional protocol are justified. PMID- 11472655 TI - Histologic analysis of clinical biopsies taken 6 months and 3 years after maxillary sinus floor augmentation with 80% bovine hydroxyapatite and 20% autogenous bone mixed with fibrin glue. AB - BACKGROUND: Bovine hydroxyapatite (Bio-Oss, Geistlich Pharmaceutical, Wollhausen, Switzerland) has been suggested to be used in maxillary sinus floor augmentation procedures prior to or in conjunction with implant placement. However, the long term histologic fate of this material is not well understood. PURPOSE: The aim with this study was to histologically evaluate the tissue response in patients to a mixture of bovine hydroxyapatite (BH), autogenous bone, and fibrin glue 6 months and 3 years after a maxillary sinus floor augmentation procedure. MATERIALS AND METHOD: Biopsies were taken from a group of 20 consecutive patients 6 months (n = 16) and 3 years (n = 12) after maxillary sinus floor augmentation with a mixture of BH (80%), autogenous bone (20%), and fibrin glue and prepared for histologic analysis. RESULTS: Light microscopy and morphometry from biopsies taken after 6 months showed various amounts of mineralized bone tissue. The specimen area was occupied by 54.1 +/- 12.6% nonmineralized tissue, followed by 21.2 +/- 24.5% lamellar bone, 14.5 +/- 10.3% BH particles, and 10.2 +/- 13.4% woven bone. The nonmineralized tissue seen in bone-forming areas consisted of a loose connective tissue, rich with vessels and cells. There were no signs of resorption of the BH particles. The lamellar bone appeared to have originated from the recipient site and was seldom in contact with the BH particles. After 3 years, the nonmineralized tissue area had decreased to 36.0 +/- 19.0% (p < .05) and consisted mainly of bone marrow tissue. The surface area of lamellar bone had increased to 50.7 +/- 22.8% (p < .05), and there was almost no immature bone. The mean specimen area occupied by BH particles, was 12.4 +/- 8.7% and had not changed from 6 months (not significant). Moreover, the sizes of the particles were similar after 6 months and 3 years. The degree of BH particle-bone contact had increased from 28.8% +/- 19.9% after 6 months to 54.5 +/- 28.8% after 3 years (p < .05). CONCLUSION: Histology of specimens from maxillary sinuses augmented with 80% BH particles, 20% autogenous bone, and fibrin glue showed a positive bone tissue response after 6 months and 3 years after augmentation of the maxillary sinus floor prior to implant placement in a group fo 20 patients. The bone surrounding and in contact with the BH particles after 6 months was mainly immature woven bone, which with time was replaced by mature lamellar bone filling the interparticle space as observed in the 3-year specimens. Moreover, bone integrated BH particles seem to be resistant to resorption. The results indicate that the procedure may be considered when only small amounts of intraoral autogenous bone graft are available. PMID- 11472656 TI - Preliminary report of immediately loaded Altiva Natural Tooth Replacement dental implants. AB - BACKGROUND: Osseointegration may be described as a direct contact between living bone and an alloplastic implant. It is thought that a period of undisturbed healing is required to ensure osseointegration. The length of the healing period is dependent on a number of factors, such as implant material, surface configuration, site preparation technique, bone quality, healing capacity of the osteotomy, and implant design. Elimination of the healing period offers distinct advantages in terms of cost of therapy and convenience to patients. Recently, a new one-piece implant design was proposed that provides improved stability of the implant to allow immediate support of an interim prosthesis. PURPOSE: This article presents the initial clinical experiences when the Altiva Natural Tooth Replacement one-piece implant was used in a human clinical trial. MATERIALS AND METHODS: This was a prospective multicenter study of the placement and immediate loading of the one-piece implant. A total of 142 implants were placed in the jaws of 93 patients. Implants were placed consecutively and were followed at specified intervals in three private clinical practices. Clinical performance was recorded relative to implant survival and prosthesis support. RESULTS: The implant survival rate was 93.7%, with similar responses in the maxilla (95.0%) and the mandible (92.7%). When implant failure occurred, it was observed as mobility without infection within 3 to 5 weeks of implant insertion. CONCLUSIONS: On the basis of this trial, there is promising evidence that this implant design can succeed at rates similar to other designs that require conventional healing times. PMID- 11472657 TI - New fabrication system for dental implant surgical stents: time-saving laboratory technique using a light-curing resin and transparent artificial teeth. AB - BACKGROUND: Surgical stents are a prerequisite for dental implant diagnosis. However, the traditional fabrication method including wax-up, investment, and resin polymerization is time consuming. PURPOSE: This article introduces a new fabrication system using a light-curing resin and transparent artificial teeth to reduce time spent in the laboratory. MATERIALS AND METHOD: The process of this system includes placing the light-curing resin and the artificial teeth on the partially edentulous dentition and alveolar ridge, making an appropriate pattern for the stent, light-curing for 5 minutes, briefly polishing, and making holes for titanium guide pins. RESULTS: This system requires only 30 to 40 minutes in the laboratory to complete a stent after mounting the casts on the articulator. Owing to the elasticity of the light-curing resin, this system eliminates the need for a blockout procedure on the undercuts of existing teeth on the casts, protects the casts from breakage, and provides the appropriate retention intraorally without any retainers. As well, titanium guide pins embedded in the stent were clearly identified by panoramic and computed tomographies. CONCLUSIONS: The new fabrication system proposed here can be time saving and, further, serve benefits for radiographic diagnosis. PMID- 11472658 TI - Osseotite implant: 3-year prospective multicenter evaluation. AB - PURPOSE: This prospective multicenter study evaluates the cumulative success rate of the Osseotite implant after 3 years of prosthetic loading. MATERIALS AND METHODS: A total of 413 Osseotite implants (Implant Innovations) were placed in 142 patients (completely or partially edentulous) in five dental offices exclusively devoted to implants. The average age of the patients was 58.3 years. Of the 413 implants, 191 were placed in the maxilla and 222 in the mandible; 271 (65.6%) were posterior implants and 142 (34.4%) were anterior implants. Clinical and radiographic evaluations were made after completion of the prosthetic restoration, after 6 months of loading, at 1 year, and at 3 years. RESULTS: A cumulative success rate of 95.3% was obtained after 3 years of prosthetic loading. The success rate was similar in both arches: 95.1% in the maxilla and 96.8% in the mandible. Early failures (before prosthetic loading) were greater (n = 12) than late failures (n = 2). After 3 years of prosthetic loading, the marginal bone level of 385 (93.2%) implants were evaluated radiographically. Bone level was at the first thread for 91.4% of the implants. A slightly increased loss was observed around 26 implants (6.7%). Including survival implants, the cumulative implant success rate after 3 years was 96%. A success rate of 98.4% was obtained with 187 short implants (8, 5 and 10 mm) reported in this multicenter evaluation. CONCLUSION: This multicenter evaluation demonstrates excellent predictability for Osseotite implants. PMID- 11472659 TI - [On recommendations, protocols, and clinical guidelines]. PMID- 11472660 TI - [Viral infection of the lower respiratory tract in hospitalized infants: etiology, clinical features and risk factors]. AB - OBJECTIVES: The aim of this study was to assess the clinical and virological characteristics of lower respiratory tract (LRT) infection in hospitalized infants in Spain and to identify clinical, radiological or laboratory parameters that could, on admission, be associated with a more severe clinical course. PATIENTS AND METHODS: A prospective study of infants less than 24 months old hospitalized for LRT infection during six consecutive seasons was performed. RESULTS: A total of 617 infants were included in the study. Diagnosis was bronchiolitis in 64 %, wheezy bronchitis in 24.6 %, laryngitis in 4.4 % and pneumonia in 6.8 %. The mean age was 269 188 days, the male/female ratio was 1:6 and the mean hospital stay was 6.6 3.5 days. At least one viral agent was identified in 55.6 % of the episodes, of which 83.6 % were due to respiratory syncytial virus (RSV). Other less frequently identified viruses were parainfluenza in 7 %, adenovirus in 4.3 % and influenza in 4 %. Coinfection was identified in 6.2 % of RSV-positive infants. These infants were younger (p < 0.005), had higher respiratory distress assessment instrument (RDAI) scores and longer hospital stay than infants in the other etiologic groups. Chest radiographs were performed in 94.3 % of the infants and 39.5 % showed infiltrate or atelectasis. This radiological alteration was significantly associated with a fever of more than 38.5 C and reactive C protein concentrations of more than 30 mg/L (p < 0.001 and p < 0.002), but not with higher RDAI score, SaO2 equal to or less than 87 %, or longer hospital stay. In the crude analysis, hospitalization for more than 5 days was associated with lower age (p < 0.01), a mean RDAI score of more than 6 (p < 0.003), SaO2 equal to or less than 87 % (p < 0.01) and RSV infection (p < 0.001). However, in the multivariate analysis only SaO2 equal to or less than 87 % and RSV infection were significantly associated with prolonged hospitalization. CONCLUSION: The most common lower respiratory tract infections in hospitalized infants in Spain are the various types of RSV-positive bronchiolitis, which have a clinical pattern similar to that described in other countries. Hypoxia on admission and RSV infection are the most important risk factors for prolonged hospitalization. PMID- 11472661 TI - [Pneumonia due to mycoplasma pneumoniae in a primary health care district]. AB - OBJECTIVE: Most studies of atypical pneumonia due to Mycoplasma pneumoniae have been performed in hospitalized patients. The aim of this study was to determine the epidemiological, clinical and radiological characteristics as well as the evolution of patients with atypical pneumonia due to Mycoplasma pneumoniae in an out-of-hospital setting. METHODS: A prospective observational study was conducted in 31 patients with atypical pneumonia due to Mycoplasma pneumoniae in the pediatric population of a primary health care district from May to July 1996. In all patients serological confirmation of infection by Mycoplasma pneumoniae was obtained using a complement-fixation test. RESULTS: The age of the children in the study ranged from 4-13 years. The most common clinical manifestations were cough (93 %), high fever (84 %) and pharyngitis (48 %). The most frequent auscultatory finding was crackles (93 %), which were bilateral in 14 patients. Clinical-radiological dissociation was found in 16 % of the children; 27 (87 %) showed radiological alterations. No characteristic radiological pattern was detected because alveolar and interstitial alterations were equally frequent, with a predominance of lung base involvement (67.7 %). In all patients response to macrolides was excellent, fever abated within 48-72 h and the remaining symptoms progressively improved. CONCLUSIONS: Atypical pneumonia due to Mycoplasma pneumoniae normally presents in children aged more than 5 years old or in adolescents. No signs, symptoms, or radiological patterns clearly indicate the etiology. However, because of its frequency, this disease should be suspected when school-aged children or adolescents present a pneumonia syndrome. Macrolides administration PMID- 11472662 TI - [Serum transferrin receptor in healthy children]. AB - BACKGROUND: Serum transferrin receptor (sTfR) is a reliable tool for assessing functional iron status and erythropoietic activity in adults, but little is known about its role in children. OBJECTIVE: To evaluate sTfR concentrations in healthy children and age and sex-related variations. A further aim was to determine the relationship of these concentrations with other iron parameters and with erythropoietin. PATIENTS AND METHODS: Blood samples from 155 healthy children aged 1-10 years old were obtained to determine sTfR, iron status indices and erythropoietin. The sTfR/ferritin ratio and the sTfR-ferritin index (sTfR/log ferritin) were calculated. Iron deficiency and increased erythropoiesis were eliminated. RESULTS: Values of sTfR, the sTfR/ferritin ratio and the sTfR-F index were 1.94 0.41mg/L (95 % CI: 1.14-2.74 mg/L), 83 40.3 (95 % CI: 4-162) and 1.4 0.36 (95 % CI: 0.69-2.1) respectively. No significant sex-related differences were found. Children aged 1-2 years old showed the highest sTfR levels. sTfR was directly related to erythrocyte count (r: 0.17; p < 0,03), hemoglobin level (r: 0.16; p < 0.05) and total iron binding capacity (r: 0.258; p < 0,001). In the multiple linear regression analysis, the best predictors of sTfR concentrations were log erythropoietin (p < 0.016) and total iron binding capacity (p < 0.0001). CONCLUSIONS: sTfR concentrations are elevated in children due to increased iron requirements for growth. In healthy children sTfR concentrations are related to erythropoietic activity and intracellular iron requirements. The current data are useful as a standard reference for healthy children. PMID- 11472663 TI - [Tobacco, alcohol and illegal drug consumption among adolescents and the relationship with lifestyle and environment]. AB - OBJECTIVES: To identify the age at which adolescents first experiment with tobacco, alcohol and illegal drugs and to identify the relationship between consumption of these substances and lifestyle and environment. METHOD: Cross sectional descriptive study based on a survey of 2,178 adolescents aged 12-16 years old from Cantabria (Spain). RESULTS: Tobacco was considered a drug by 80.4 % of the adolescents. Forty-four percent had smoked at some time and 19.3 % were smokers. Alcohol was not considered a drug by 44.5 %, 69.2 % had drunk alcohol at some time and 37 % drank regularly. Most of the adolescents (92.9 %) drank at weekends and 88 % drank all the spirits surveyed. The adolescents were introduced to this habit by friends (54.1 %) and relatives (16.4 %). Forty-six percent of those who drank regularly had got drunk once or more in the previous 6 months. At the age of 15 years, 29.4 % smoked and 61.8 % drank alcohol regularly. An association with illegal drugs was less common; 10.2 % smoked hashish. Consumption of all these substances was associated with greater age, living in an environment in which these substances were consumed and with certain forms of leisure activity. Logistic regression analysis revealed that considering tobacco and alcohol as drugs protected adolescents against their consumption and that consuming other drugs, getting drunk and being in a consumer environment were risk factors for consumption. CONCLUSIONS: Contact with illegal drugs, alcohol and tobacco starts early in adolescence. Levels of consumption are worrying. Environment and lifestyle are closely associated with risk factors for consumption. PMID- 11472664 TI - [Pre-surgical worries: an empirical study in the child and adolescent population]. AB - OBJECTIVE: To analyze presurgical worries in children and adolescents and to examine differences according to gender and prior surgical experience. METHOD: The child sample included 1,311 subjects, aged 7-10 years old, and the (pre)adolescent sample included 1,488 subjects, aged 11-14 years old. The child sample filled in the child version of the Surgical worries Questionnaire (CPCI-N; Quiles, Ortigosa and Mendez, 1998), with 17 items and a three-point rating scale, and the adolescent sample filled in the adolescent version (CPCI-A, Quiles, Mendez and Ortigosa, 1998), with 22 items and a five-point rating scale. Student's t-test was carried out to analyze the main worries and an analysis of variance was used to examine the effect of gender. RESULTS: Children's and adolescents' main worries concerned the surgical process, the negative results of surgery and the negative consequences of illness. However, the younger children showed other significant worries such as having a needle in the arm for a long time or separating from their parents. In both samples, girls and subjects without previous experience of surgery showed higher levels of pre-surgical worries. CONCLUSIONS: We discuss the implications of these results for the development of programs of psychological preparation for children and adolescents about to undergo surgery. PMID- 11472665 TI - [International adoption: information guide and medical evaluations]. AB - This article aims to provide a guide to pediatricians attending families involved in international adoptions. Firstly, we describe the pre-adoption consultation, which includes a review of the information available on the child, the immunizations and medications recommended for traveling to the foreign country, and the information concerning the more frequent health problems observed in children adopted from abroad. Secondly, we describe the post-adoption evaluation, which includes a physical examination and the laboratory screening tests recommended for internationally adopted children. PMID- 11472666 TI - [Recommendations for the care of healthy newborn infants]. AB - This article makes certain recommendations on the care of the healthy newborn. Firstly, we discuss the situations that should be reported to the pediatrician/neonatologist and the reasons why the presence of these specialists is required in the delivery room (urgent or elective cesarean section, preterm labor). Secondly, we discuss the most important guidelines to follow in the delivery room and after birth. Concerning care in the delivery room, we stress the importance of care of the newborn (especially of the umbilical cord), bonding between the mother and child, identification of the newborn, assessment of neonatal adaptation to extrauterine life, prevention of ophthalmia neonatorum and hypoprothrombinemia, placing the baby correctly in the crib and hepatitis B prophylaxis. Concerning the postnatal period, we recommend feeding (promotion of breast feeding), rooming-in with the mother if the newborn is hospitalized in the nursery screening for hypoacousia and metabolic diseases, and discharge with special surveillance in cases of early discharge. PMID- 11472667 TI - [Recommendations for perinatal transport]. AB - Perinatal transport should be integrated into a system of perinatal care within a regional health care program and should be planned according to the healthcare map of each community. We describe the various types of transport, their advantages and disadvantages, the resources required, and the protocol that should be followed in perinatal transfer. We highlight the importance of maternal and neonatal transport. The organization of transfers receives special attention, and we discuss the different functions of the coordinating, referral and receiving centers as well as those of the transport assistance team. We also discuss ethical-legal questions. PMID- 11472668 TI - [Eosinophilic pustular folliculitis in childhood]. AB - BACKGROUND: Eosinophilic pustular folliculitis (EPF) is an inflammatory disorder of unknown etiology. In infants this disorder is characterized by recurrent episodes of sterile pustules primarily or exclusively involving the scalp with occasional involvement of the face, trunk and extremities. There are few reports of EPF in children. OBJECTIVE: To describe the clinical features and evolution of four pediatric patients and to discuss the main differential diagnoses. METHODS: Biopsy specimens were examined, pustules were cultured and laboratory tests were analyzed. RESULTS: Four patients (3 males and 1 female) aged 7-18 months presented with self limiting recurrent pruritic papules and pustules on the scalp. In one patient, the lesions were mainly localized on the extremities. Cultures for bacteria, fungi and viruses were negative. No systemic disease was found. Topical steroids were effective in three patients but pustules recurred after treatment was stopped. Cetirizine and Hydroxacen were administered in two corticoid-resistant patients with fair response. No other systemic therapy was administered. Peripheral eosinophilia was detected in three patients. CONCLUSIONS: EPF in infants seems to be a clearly defined entity. Although few cases have been described in children, this dermatosis is undoubtedly more frequent than suggested by the literature. Consequently, pediatricians should be aware of its existence. PMID- 11472669 TI - [Whitish lesions in oral mucosa]. AB - Whitish lesions in oral mucosa are usually diagnosed as candidosis, lichen planus or dysplasias. However, other infrequent entities associated with leukoplakia should be borne in mind. We present the case of a 7-year-old girl with whitish lesions in the oral mucosa. Laboratory, mycological and histopathological investigations were performed and the patient was diagnosed with oral white sponge nevus. PMID- 11472670 TI - [Peutz-Jegher's syndrome]. AB - Peutz-Jeghers syndrome is a rare hereditary disease, although in about 20 % of patients there is no known family history. Its clinical hallmarks are facial mucocutaneous pigmentation and diffuse gastrointestinal polyposis of hamartomatous origin. The major difficulty in the management of this disease lies in the complications of surgery for small bowel polyposis and the high incidence of tumors presented by these patients as adults. We present the case of a 10-year old girl with treatment-resistant anemia and no other clinical symptoms. Further investigation revealed pigmentation in the oral mucosa and polyposis in the stomach and small intestine. Hamartomatous lesions were confirmed by histological study of intestinal biopsy. No familial antecedents of pigmentation or intestinal polyps were found. During evolution the patient required subtotal resection of the small intestine due to invagination. PMID- 11472671 TI - [Metastatic Crohn's disease in childhood]. AB - Crohn's disease (CD) is an intestinal inflammatory disease of unknown origin that is sometimes associated with cutaneous manifestations. These skin lesions are usually nonspecific (erythema nodosum, erythema multiforme and pyoderma gangrenosum) and should be differentiated from the cutaneous lesions corresponding to the typical histological pattern of CD; most of these extend directly from the involved bowel. Noncaseating granulomatous infiltration in skin anatomically distant from areas of gastrointestinal CD and separated by normal skin is extremely rare. This entity, known as metastatic Crohn's disease, can be the first symptom of CD or even more unusually can appear during the course of the disease.We report the case of a 9-year-old boy, previously diagnosed with ileocolic and perianal CD, who had been receiving treatment with mesalazine, prednisone and 6-mercaptopurine for 3 years. He presented with swelling of the penis and scrotum of 2 weeks' evolution, without any other symptoms. Testicular ultrasound showed thickening of the scrotal wall and increased flux at this site. Skin biopsy showed noncaseating granulomas. Results of special staining techniques were negative for the presence of microorganisms. The prednisone dose was increased and metronidazole was added, with noticeable improvement in the lesions. Due to reappearance of cutaneous lesions when the prednisone dose was reduced, the patient currently receives maintenance treatment. PMID- 11472672 TI - [Acute abdominal pain in a 10-year-old girl]. PMID- 11472673 TI - [Cholelithiasis]. PMID- 11472675 TI - [Cerebral abscess provoked by Pseudomonas aeruginosa. Complication of ethmoiditis in a healthy two-month-old infant]. PMID- 11472676 TI - [Intracraneal epidural abscess as a complication of sinusitis]. PMID- 11472677 TI - [Incontinencia pigmenti associated with Klinefelter's syndrome]. PMID- 11472678 TI - [Generalized granuloma annulare. Report of two cases]. PMID- 11472682 TI - [Molecular analysis of the RPE65 gene in 72 Spanish families with autosomal recessive retinitis pigmentosa]. AB - BACKGROUND: Autosomal recessive retinitis pigmentosa (arRP) is the most common form of retinitis pigmentosa (RP). It is characterized by a high degree of allelic and non-allelic genetic heterogeneity. Previously,it has been demonstrated that the RPE65 gene is responsible for 2% recessive or isolated RP cases and 16% Leber's congenite amaurosis cases. Although the concrete function of RPE65 is unknown as yet, it has been found to be involved in vitamin A metabolism and rhodopsin regeneration. PATIENTS AND METHOD: We studied the involvement of the RPE65 gene in 72 arRP Spanish families by means of indirect molecular and mutation analysis. RESULTS: The results obtained using the intragenic microsatellite marker D1S2803 allowed us to exclude RPE65 as the causative gene of the disease in 80.5% of the families studied. Three new variants of the RPE65 gene have been identified: IVS6-43delA, IVS6-42delT and IVS6-33CG. We found that IVS6-33C-->G was a common polymorphism. The other variants, namely IVS6-43delA and IVS6-42delT, were not identified in 150 control chromosomes studied. The segregation analysis of IVS6-42delT variant seemed to exclude it as being involved in the disease. CONCLUSIONS: Our results argue against the involvement of RPE65 gene in the families studied, indicating that the prevalence of RPE65 abnormalities in arRP Spanish families may be lower than that observed in other populations. PMID- 11472683 TI - [Treatment with LHRH analogues in girls with precocious puberty does not improve final height. Longitudinal study compared with a control group]. AB - BACKGROUND: We analysed the effectiveness of therapy with LHRH analogues in girls with a puberty onset at age 8 years. PATIENTS AND METHOD: We performed a non randomised clinical study of 32 girls with advanced puberty. These included 16 treated with triptorelin LHRH analogue(3.75 mg/month during 1 year) and 16 control subjects. We carried out anthropometric measurements and determined the pubertal height growth (gain in height from the puberty onset up to the final height) and the pubertal duration (time in years from the puberty onset up to the age at which final height is attained). RESULTS: Treatment with LHRH analogue delayed the menarche age (11.5 [1.46]vs 10.37 [0.67] years of age; p = 0.03), led to an involution in secondary sexual characteristics and a temporary decrease ingrowth rate, and delayed skeletal maturation. However, pubertal duration, pubertal height growth and final height were all similar in both groups. In addition, no significant differences in body fat mass were observed. CONCLUSIONS: Treatment with LHRH analogues in advanced puberty modifies pubertal development, without modifying pubertal duration or pubertal height growth. Furthermore, this treatment does not improve final height. PMID- 11472684 TI - [Spanish language adaptation and validation of the Pfeiffer's questionnaire (SPMSQ) to detect cognitive deterioration in people over 65 years of age]. AB - BACKGROUND: Our aims were to develop a Spanish version of the short portable mental status questionnaire (SPMSQ) (Pfeiffer's test) and to validate the resultant Spanish version of the test among a general population aged 65 years or older. PATIENTS, MATERIAL AND METHODS: First, we developed the Spanish version of the SPMSQ by means of a cross-cultural adaptation methodology. Patients were the assessed by independent teams in two sessions the same day. The first team (two trained psychiatrists and one trained family physician) carried out a standardised neuropsychological assessment to diagnose cognitive impairment/dementia according to ICD-10 criteria which was regarded as the reference "gold standard" to calculate validity parameters (sensitivity and specificity). The second team (family physicians/nurses) assessed the presence of cognitive impairment with the Spanish version of the SPMSQ. RESULTS: 255 subjects (66.7%, females) underwent the assessment. Mean age was 74.5 years and 65.5% of them were illiterate. The inter-rater reliability and test-retest reliability of the SPMSQ Spanish version were 0.738 (p < 0.001), and 0.925 (p < 0.001), respectively,with a convergent validity of 0.74 (p < 0.001) and a discriminant validity of 0.230 (p < 0.001). The internal consistency was high with an *value of 0.82. The area under the ROC curve was 0.89. Sensitivity and specificity were 85.7 and 79.3, respectively (cut-off of 3 or more). CONCLUSIONS: The Spanish version of the SPMSQ test has a good reliability and validity. For clinical use, a cut-off of 3 appears to be most useful. When illiterate patients are assessed, the it is recommended to employ a cut-off of 4 or more. PMID- 11472685 TI - [Retinitis pigmentosa]. PMID- 11472686 TI - [New therapeutical approaches to type 2 diabetes]. PMID- 11472687 TI - [Familial Mediterranean fever]. PMID- 11472688 TI - [New induction immunosuppression treatments in kidney transplantation]. PMID- 11472689 TI - [Breast cancer in Rubens and Rembrandt paintings]. PMID- 11472691 TI - [Necrolytic migratory erythema as initial manifestation of normoglycemic glucagonoma]. PMID- 11472692 TI - [Churg-Strauss syndrome associated with montelukast therapy]. PMID- 11472694 TI - Bundle branch block as a predictor of long-term survival after acute myocardial infarction. AB - Using a community-based population of patients with acute myocardial infarction (AMI), we sought to: (1) determine the prevalence of bundle branch block (BBB) on the presenting electrocardiogram (ECG), (2) compare the clinical characteristics and the treatment administered to patients with and without BBB, and (3) determine the association of BBB with mortality. We analyzed the admission ECGs of 894 consecutive patients with AMI from Olmsted County, Minnesota, seen at our institution from January 1988 to March 1998. Of these, 53 had left BBB (LBBB) (5.9%) and 60 had right BBB (RBBB) (6.7%). Patients with BBB were more likely to be older, have a history of AMI or hypertension, and to be in Killip class >I at presentation. They were less likely to receive primary reperfusion therapy, beta blockers, or heparin, but more likely to receive angiotensin-converting enzyme inhibitors. They had lower mean predischarge ejection fractions (38 +/- 16% vs 50 +/- 15%, p <0.0001). In-hospital mortality was 13.3%, 17.0%, and 9.1% for patients with RBBB, LBBB, and no BBB, respectively (p = 0.11). Respective postdischarge survival at 1, 3, and 5 years was 80%, 60%, and 50% in the RBBB group, 78%, 56%, and 51% in the LBBB group, and 92%, 85%, and 76% in the group without BBB (p <0.0001). Although BBB was not an independent predictor of mortality on multivariate analysis, the presence of transient or persistent BBB with AMI is an easily recognized clinical marker of increased mortality. Our conclusion from this study is that in a community-based population, patients who had LBBB or RBBB at the time of AMI had lower predischarge ejection fractions and higher in-hospital and long-term unadjusted mortality. PMID- 11472695 TI - Determinants and prognostic implications of terminal QRS complex distortion in patients treated with primary angioplasty for acute myocardial infarction. AB - Terminal QRS complex distortion on admission has an impact on a patient's prognosis after primary angioplasty for acute myocardial infarction (AMI). We evaluated the determinants and prognostic significance of terminal QRS complex distortion in 153 consecutive patients with AMI after primary angioplasty. The study population was divided into 2 groups according to the presence (group I, n = 41) or absence (group II, n = 112) of terminal QRS complex distortion. The primary end points were the occurrence, within 6 weeks after AMI, of death, nonfatal reinfarction, or congestive heart failure. Baseline characteristics were similar between the 2 groups. However, patients in group I had higher peak levels of serum creatine kinase than those in group II (5,100 +/- 3,100 vs 3,000 +/- 1,800 U/L, respectively, p <0.01). The rate of angiographic no-reflow (Thrombolysis In Myocardial Infarction flow grade < or =2) was 31.7% in group I and 10.7% in group II (p <0.01). The predischarge left ventricular ejection fraction was 45.0 +/- 12.0% in group I and 54.0 +/- 8.0% in group II (p <0.01). Multivariate analysis identified the pressure-derived fractional collateral flow index and the culprit lesion in the left anterior descending coronary artery as independent determinants of the terminal QRS complex distortion. No patients died during 6 weeks of follow-up. The 2 groups were similar for life-threatening arrhythmia or reinfarction. However, there were more patients in group I than in group II with congestive heart failure (26.8% vs 5.4%, respectively, p <0.01) or who reached the primary end points (29.3% vs 5.4%, respectively, p <0.01). In conclusion, terminal QRS complex distortion on admission is associated with poor clinical outcome after primary angioplasty for AMI, and collateral flow may have a major influence on terminal QRS complex distortion during AMI. PMID- 11472696 TI - Chlamydia pneumoniae exposure and inflammatory markers in acute coronary syndrome (CIMACS). AB - Previous studies have shown higher levels of Chlamydia pneumoniae (C. pneumoniae, CP) antibody titers (CPIgG), C-reactive protein (CRP), and fibrinogen in patients with coronary artery disease. The role of these infectious and inflammatory markers in precipitating acute coronary syndrome (ACS) is unclear. We conducted a cross-sectional study on patients (n = 830, mean age 63 +/- 15 years, 57% male) admitted to the chest pain center of our institution. The differences in the CPIgG, CRP, and fibrinogen levels in patients who were diagnosed with ACS versus those who were not (non-ACS) were evaluated. CPIgG titers tended to be higher in the ACS group than in the non-ACS group. However, when different titers were used to define seropositivity, the difference achieved statistical significance only at the titer of > or =1:1,024 (35% vs 26%, p = 0.004). CRP (median 0.48 vs 0.33 mg/dl, p <0.0001), fibrinogen (median 317 vs 293 mg/dl, p <0.0001), and leukocyte count (median 7.7 vs 6.9 10(9)/L, p <0.0001) were higher in the ACS group. On multivariate analysis, CPIgG > or =1:1,024 (odds ratio [OR] 1.62), diabetes (OR 1.91), hypertension (OR 1.46), prior myocardial infarction (OR 1.78), smoking (OR 1.70), Caucasian race (OR 1.7), high-density lipoprotein (OR 0.98), and elevated troponin-T (OR 12.44) were the only factors independently associated with ACS. Thus, we found a strong association between high level seropositivity to CP and ACS. This may indicate recent re-infection or an exaggerated immune response to CP as an etiologic factor for ACS. This study also suggests that therapeutic interventions may need to be specifically targeted to these patients. PMID- 11472697 TI - Usefulness of calcium scoring using electron beam computed tomography and noninvasive coronary angiography in patients with suspected coronary artery disease. AB - The aim of this study was to investigate the reliability of calcium scoring (CS) and electron beam computed tomographic angiography (EBCTA) as a noninvasive tool in the diagnosis of coronary artery disease (CAD): 93 consecutive patients (aged 59 +/- 9 years) with symptoms suspicious for CAD underwent CS. In 87 of these subjects, an additional EBCTA investigation was performed. Using receiver operating characteristic curve analysis, we determined a calcium score cut point providing an overall sensitivity of 80% and a specificity of 72% in detecting patients with CAD. For clinical purposes the use of cut points is difficult. We therefore determined score ranges providing >80% specificity (high score range) and >85% sensitivity (low score range) and determined the scores between these ranges as equivocal borderline scores. Calculated on a per-segment basis in assessable proximal and midcoronary segments, the sensitivity for detecting coronary stenoses >50% was 78%, and the specificity was 93%. Thus, 32 of 44 patients with significant CAD and 24 of 49 patients without CAD were correctly classified. The combination of CS and EBCTA predicted CAD in 77% (72 of 93) of patients. No or low calcium scores provided high specificity for ruling out CAD. The addition of EBCTA in those patients improved sensitivity. In patients with high calcium scores, accuracy of EBCTA was not significantly different from CS alone (72% vs 83%), whereas in patients with borderline scores it was significantly superior (80% vs 58%, p <0.03). Thus, the complementary use of CS and EBCTA appears beneficial, particularly in patients with borderline scores, and could improve sensitivity in the low score range. In the presence of high scores, no major diagnostic gain from an additional EBCTA versus CS alone could be observed. PMID- 11472698 TI - Usefulness of magnesium sulfate in stabilizing cardiac repolarization in heart failure secondary to ischemic cardiomyopathy. AB - Experimental heart failure is associated with cardiac magnesium loss, causing increased beat-to-beat variability in the action potential. Unstable repolarization contributes to sudden death, but no therapy has been shown to reduce repolarization variability safely. We sought to test whether a prolonged infusion of magnesium sulfate (MgSO(4); 40 mmol/24 hours) would normalize QT interval variability in patients with compensated heart failure. Fifteen patients (New York Heart Association class II to III; mean age 63 years) were enrolled in a placebo-controlled, double-blind study. Surface electrocardiograms were recorded and digitized at entry and at 48 and 168 hours (drug washout). Repolarization stability was assessed using an automated method measuring each QT interval in a 5-minute epoch. The QT variability index was derived as the ratio of normalized QT-to-normalized heart rate variability. Seven of 15 patients received MgSO(4). Mean heart rate and QT did not change in either group. The QT variability index was stable in the placebo group (-0.69 +/- 0.15 at entry, -0.71 +/- 0.22 at 48 hours, -0.70 +/- 0.18 at 168 hours), but decreased significantly in the treated group at 48 hours (-0.95 +/- 0.19 to -1.36 +/- 0.13, p <0.05 repeated-measures analysis of variance), returning to baseline at 168 hours ( 0.84 +/- 0.18). Regression analyses showed that administration of MgSO(4) resulted in a stronger correlation between the QT and RR interval (p <0.01). Thus, MgSO(4) stabilizes cardiac repolarization in patients with compensated heart failure due to ischemic heart disease. Magnesium therapy may be useful in altering the proarrhythmic substrate in heart failure. PMID- 11472699 TI - Profile and prevalence of aspirin resistance in patients with cardiovascular disease. AB - We determined the prevalence and clinical predictors of aspirin resistance by prospectively studying 325 patients with stable cardiovascular disease who were receiving aspirin (325 mg/day for > or =7 days) but no other antiplatelet agents. We also compared the detection of aspirin resistance with optical platelet aggregation, a widely accepted method, with a newer, more rapid method, the platelet function analyzer (PFA)-100, a whole blood test that measures platelet adhesion and aggregation ex vivo. Blood samples were analyzed in a blinded fashion for aspirin resistance by optical aggregation using adenosine diphosphate (ADP) and arachidonic acid, and by PFA-100 using collagen and/or epinephrine and collagen and/or ADP cartridges to measure aperture closure time. Aspirin resistance was defined as a mean aggregation of > or =70% with 10 microM ADP and a mean aggregation of > or =20% with 0.5 mg/ml arachidonic acid. Aspirin semiresponders were defined as meeting one, but not both of the above criteria. Aspirin resistance by PFA-100 was defined as having a normal collagen and/or epinephrine closure time (< or =193 seconds). By optical aggregation, 5.5% of the patients were aspirin resistant and 23.8% were aspirin semiresponders. By PFA 100, 9.5% of patients were aspirin resistant. Of the 18 patients who were aspirin resistant by aggregation, 4 were also aspirin resistant by PFA-100. Patients who were either aspirin resistant or aspirin semiresponders were more likely to be women (34.4% vs 17.3%, p = 0.001) and less likely to be smokers (0% vs 8.3%, p = 0.004) compared with aspirin-sensitive patients. There was a trend toward increased age in patients with aspirin resistance or aspirin semiresponders (65.7 vs 61.3 years, p = 0.06). There were no differences in aspirin sensitivity by race, diabetes, platelet count, renal disease, or liver disease. PMID- 11472700 TI - Oral glycoprotein IIb/IIIa antagonism in patients with coronary artery disease. AB - Dose-finding studies and trials of interaction of oral glycoprotein IIb/IIIa antagonists with other antiplatelet agents have been limited. We hypothesized that these detailed assessments could be first performed in patients with stable coronary artery disease (CAD) and then extrapolated to the target population. To this end, we performed 2 sequential studies. The first study examined the dose related effects on indexes of platelet and vascular function induced by the oral inhibitor RPR 109891, when given alone and in combination with aspirin, in patients (n = 100) with stable CAD. The second study (the Antagonism of the FIbrinogen Receptor after Myocardial Events trial) assessed the pharmacodynamics and safety of derived regimens in patients (n = 320) with unstable coronary syndromes. In patients with stable CAD, platelet aggregation was dose dependently inhibited by RPR 109891, and the dose-response relation was shifted to the right by the concomitant administration of aspirin (p = 0.0001). The degree of platelet inhibition induced by 3 doses of RPR 109891 (plus aspirin) was lower in patients with unstable than stable CAD. No drug-related major bleeding occurred in either study. RPR 109891 treatment was associated with acute and delayed thrombocytopenia. In conclusion, chronic treatment with an oral glycoprotein IIb/IIIa antagonist (1) induces antiplatelet effects that are potentiated by concomitant administration of aspirin, (2) may require dose adjustment in syndromes of platelet activation, (3) is associated with a low rate of clinically significant bleeding when doses inducing incomplete inhibition of platelet aggregation are used, and (4) requires frequent monitoring of platelet count unless reliable predictors of delayed thrombocytopenia become available. PMID- 11472701 TI - Role of vessel size as a predictor for the occurrence of in-stent restenosis in patients with diabetes mellitus. AB - Intracoronary stents have been shown to reduce the rate of restenosis when compared with balloon angioplasty, but in-stent restenosis continues to be an important clinical problem. It was therefore the aim of this registry to identify procedural and angiographic predictors for the occurrence of in-stent restenosis. We analyzed 368 patients with 421 lesions who underwent coronary stent implantation between January 1998 and February 2000. Indications for the placement of a coronary stent were severe dissections (37%), suboptimal angiographic results (38%), restenotic lesions (20%), and graft lesions (4%). Angiographic follow-up was obtained in 270 patients (73%) with 293 lesions after 6 months. Clinical and angiographic variables were analyzed by univariate and multivariate models for the ability to predict the occurrence of in-stent restenosis, defined as a diameter stenosis >50%. In-stent restenosis was angiographically documented in 67 patients and 68 lesions (23%). Under all tested variables the reference luminal diameter before stent implantation (p = 0.006) and diabetes mellitus (p = 0.023) were identified as independent predictors for the occurrence of in-stent restenosis. The comparison of diabetic and nondiabetic patients according to vessel size revealed a 2 times higher rate of in-stent restenosis in small vessels (44% vs 23%, p = 0.002), whereas in vessels >3.0 mm the rate of in-stent restenosis was not significantly different between the 2 groups. In this registry, the clinical variable diabetes and the procedural variable reference vessel size were independent predictors for the occurrence of in-stent restenosis. In these patients, the rate of in-stent restenosis was as high as 45%. PMID- 11472702 TI - Extent and distribution of in-stent intimal hyperplasia and edge effect in a non radiation stent population. AB - Intimal hyperplasia within the body of the stent is the primary mechanism for in stent restenosis; however, stent edge restenosis has been described after brachytherapy. Our current understanding about the magnitude of in vivo intimal hyperplasia and edge restenosis is limited to data obtained primarily from select, symptomatic patients requiring repeat angiography. The purpose of this study was to determine the extent and distribution of intimal hyperplasia both within the stent and along the stent edge in relatively nonselect, asymptomatic patients scheduled for 6-month intravascular ultrasound (IVUS) as part of a multicenter trial: Heparin Infusion Prior to Stenting. Planar IVUS measurements 1 mm apart were obtained throughout the stent and over a length of 10 mm proximal and distal to the stent at index and follow-up. Of the 179 patients enrolled, 140 returned for repeat angiography and IVUS at 6.4 +/- 1.9 months and had IVUS images adequate for analysis. Patients had 1.2 +/- 0.6 Palmaz-Schatz stents per vessel. There was a wide individual variation of intimal hyperplasia distribution within the stent and no mean predilection for any location. At 6 months, intimal hyperplasia occupied 29.3 +/- 16.2% of the stent volume on average. Lumen loss within 2 mm of the stent edge was due primarily to intimal proliferation. Beyond 2 mm, negative remodeling contributed more to lumen loss. Gender, age, vessel location, index plaque burden, hypercholesterolemia, diabetes, and tobacco did not predict luminal narrowing at the stent edges, but diabetes, unstable angina at presentation, and lesion length were predictive of in-stent intimal hyperplasia. In a non-radiation stent population, 29% of the stent volume is filled with intimal hyperplasia at 6 months. Lumen loss at the stent edge is due primarily to intimal proliferation. PMID- 11472703 TI - Comparison of self-expanding and balloon-expandable stents for the reduction of restenosis. AB - To compare the efficacy of self-expanding (SE) and balloon-expandable (BE) stents in native coronary arteries, we randomly assigned 1,096 patients with new and restenotic lesions to receive either device. Baseline demographics and coronary angiographic characteristics were similar in the 2 groups. The incidence of major adverse cardiac events including death, myocardial infarction, bypass surgery, and repeat intervention was similar for both groups at 1 month (2.9% vs 3.1% for SE vs BE, respectively) and at 9 months (19.3% vs 20.1%, SE vs BE respectively). In a subgroup of patients who underwent follow-up angiography (n = 250), the binary restenosis rates (24.2% vs 18.7%, p = 0.30), late loss (0.98 vs 94 mm, p = 0.60), and loss index (0.55 vs 55, p = 0.95) were not significantly different for both groups. In 62 patients who underwent intravascular ultrasound examination (IVUS), there was a trend toward a lower incidence of edge tears in the SE group (6% vs 23%, p = 0.06). Follow-up IVUS analysis showed that the minimum stent area of the SE stent increased by 33% at 6 months, whereas no change occurred in the BE stents; this was accompanied by a greater degree of intimal proliferation in the SE stents compared with BE stents (3.1 +/- 2.0 vs 1.7 +/- 1.7 mm(2)). Thus, the SE stents had similar clinical and angiographic outcomes in patients with lesions in native coronary arteries. PMID- 11472704 TI - Metabolic disorders contribute to subclinical coronary atherosclerosis in patients with coronary calcification. AB - This investigation determined the prevalence of low-density lipoprotein (LDL) subclass distribution abnormalities, elevated lipoprotein(a) (Lp(a)), and elevated total plasma homocysteine in asymptomatic subjects with subclinical coronary artery disease determined by electron beam tomography (EBT). Fifty-five percent of subjects were classified as higher risk patients and 45% as lower risk patients, employing the National Cholesterol Education Program (NCEP) lipid criteria. EBT was performed in 296 consecutive asymptomatic subjects, and blood was analyzed for total, LDL, and high-density lipoprotein (HDL) cholesterol, triglycerides, LDL subclass distribution by S(3) gradient gel electrophoresis, Lp(a), and total homocysteine. Disorders of LDL subclass distribution were the most common disorder with 60.6% of the population expressing a distribution in the small regions IIIa + IIIb of >20%; and this was more common in the NCEP higher risk group (LDL cholesterol > or =130 and/or HDL cholesterol <35 mg/dl) (p <0.0004). A Lp(a) value >25 mg/dl was found significantly more often in the NCEP higher (36.9%) compared with lower (14.3%) risk group (p <0.001). None of the laboratory measurements correlated with the calcium score or calcium score percentile rank, with the exception of a weak correlation of mean LDL peak particle diameter and calcium percentile (r = 0.14, p = 0.02). Determination of metabolic disorders in addition to LDL cholesterol and HDL cholesterol increased the diagnostic yield from 55.1%, based on NCEP lipid criteria, to 84.1% with the addition of LDL subclass distribution, Lp(a), and total homocysteine. We conclude that: (1) disorders of LDL subclass distribution and elevated Lp(a) occur frequently in NCEP higher risk patients with subclinical coronary artery disease and are the only identifiable disorders in lower NCEP risk patients; and (2) electron beam tomographic evaluation and determination of LDL subclass distribution and Lp(a) should be considered for incorporation into primary prevention guidelines. PMID- 11472705 TI - Correlation of non-high-density lipoprotein cholesterol with apolipoprotein B: effect of 5 hydroxymethylglutaryl coenzyme A reductase inhibitors on non-high density lipoprotein cholesterol levels. AB - Apolipoprotein B has been shown to be a better predictor of coronary heart disease than low-density lipoprotein (LDL) cholesterol, and non-high-density lipoprotein (non-HDL) cholesterol may also be a better parameter for coronary heart disease risk assessment and as a target for therapy. Data from the Atorvastatin Comparative Cholesterol Efficacy and Safety Study (ACCESS) were used to assess the correlation between lipid and apolipoprotein B levels before and after lipid-lowering therapy and to examine the effects of 5 hydroxymethylglutaryl coenzyme A reductase inhibitors on lipids and apolipoprotein B. The 54-week study randomized 3,916 hypercholesterolemic patients to atorvastatin, fluvastatin, lovastatin, pravastatin, or simvastatin, initiated at recommended starting doses with titrations as needed at weeks 6, 12, and 18 to achieve National Cholesterol Education Program LDL targets. Compared with LDL cholesterol, non-HDL cholesterol correlated better with apolipoprotein B levels at baseline (r = 0.914, p <0.0001) and at week 54 (r = 0.938, p <0.0001), and the correlation was strong across all baseline triglyceride strata. At starting doses, atorvastatin (10 mg) lowered non-HDL cholesterol by 33.3% compared with 26.6% with simvastatin (10 mg), 24.1% with lovastatin (20 mg), 17.2% with fluvastatin (20 mg), and 17.0% with pravastatin (10 mg). Atorvastatin also provided greater reductions in non-HDL cholesterol after dose titration, and a greater percentage of patients taking atorvastatin achieved non-HDL cholesterol targets. Baseline triglyceride did not affect non-HDL cholesterol reductions with any of the 5 hydroxymethylglutaryl coenzyme A reductase inhibitors. Fewer patients achieved non-HDL cholesterol targets than LDL cholesterol targets, particularly among high-risk patients, implying that if non-HDL cholesterol was used as a target for treatment, more patients would need to be treated more aggressively than National Cholesterol Education Program guidelines require. PMID- 11472706 TI - Effect of niacin and atorvastatin on lipoprotein subclasses in patients with atherogenic dyslipidemia. AB - This study was conducted to determine the efficacy of atorvastatin and niacin on lipoprotein subfractions in patients with atherogenic dyslipidemia. This was a multicenter, randomized, open-label, parallel-design study of patients with total cholesterol >200 mg/dl, triglycerides between 200 and 800 mg/dl, and apolipoprotein B >110 mg/dl. Patients were randomly assigned to atorvastatin 10 mg or immediate release niacin 3,000 mg daily for 12 weeks following a low-fat diet stabilization period. Lipoprotein subclasses were measured by nuclear magnetic resonance spectroscopy. Atorvastatin and niacin both significantly reduced the concentrations of very low-density lipoprotein (VLDL) particles (-31% and -29%, respectively) and small low-density lipoprotein (LDL) particles (-44% and -35%, respectively). Niacin increased the concentration of large LDL (+75%). Atrovastatin reduced the number of LDL particles more than niacin (31% vs 14%). In patients with atherogenic dyslipidemia, both drugs had important effects on lipoprotein subfractions, which contributed to a reduction in coronary heart disease risk. The drugs equally reduced VLDL subclass levels. Niacin shifted the LDL subclass distribution toward the larger particles, more effectively converted patients from LDL phenotype B to phenotype A, and increased levels of the larger and perhaps more cardioprotective high-density lipoprotein particles. In contrast, atorvastatin preferentially lowered the concentration of small LDL particles without increasing levels of large LDL, and more effectively, reduced LDL particle numbers. Atorvastatin had a preferred LDL effect, whereas niacin had a preferred high-density lipoprotein effect. PMID- 11472707 TI - Relation between myocyte disarray and outcome in hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HC) is associated with an increased risk of sudden cardiac death or death from heart failure. Little is known of the pathologic substrate for risk of premature death in this disease. We therefore set out to correlate the pathologic findings with the mode of death and risk profile in 75 patients with HC. Hearts with HC were obtained after death or transplantation. The clinical details were correlated with the macroscopic findings and the percent fibrosis, disarray, and small-vessel disease across 19 sections of each heart. Thirty-nine patients died suddenly, 28 had end-stage heart failure, and 8 died of other causes. Myocyte disarray correlated positively with evidence of ischemia (r = 0.5, p <0.0001), and was greater in patients who died before age 21 years (mean disarray 33% vs 18%, p <0.0001) and in those with an abnormal vascular response to exercise (mean disarray and 30% vs 19%, p = 0.04). Myocardial fibrosis was greater in patients who died in heart failure (mean percent fibrosis was 2.8% versus 0.9%, p = 0.003), and in patients with nonsustained ventricular tachycardia or a high risk fractionation study (4.9% vs 2.7%, p = 0.04, and 6.84% vs 2.8%, p = 0.03, respectively). In conclusion, young patients who die with HC have greater disarray than their older counterparts. In contrast, myocardial fibrosis is the substrate for premature deaths from heart failure and is associated with an increased risk of a primary ventricular arrhythmia. PMID- 11472708 TI - Increased incidence of atrial flutter associated with the rejection of heart transplantation. AB - Atrial fibrillation (AF) and atrial flutter (Afl) are common dysrhythmias that occur after orthotopic heart transplantation (OHT); however, their etiology and clinical significance have not been defined. To determine the precise incidence of sustained AF and Afl and their association with cardiac rejection, 892 consecutive patients who underwent OHT were studied. A total of 104 patients had 113 episodes of Afl; 102 patients had 117 episodes of AF. The incidence of Afl (12.7%) was the same as AF (13.1%). Sixty-nine AF episodes occurred in first 2 weeks after transplantation, and 22 of which were associated with rejection. In contrast, only 20 Afl episodes occurred the first 2 weeks after OHT, 10 of which were associated with rejection. Fifty-two episodes of Afl occurred during from the third week to 6 months after transplantation, 34 of which were associated with moderate to severe cellular or humoral rejection and/or transplant coronary artery disease (TCAD). All 41 Afl episodes that occurred 6 months after transplantation were associated with cellular and humoral rejection, and/or TCAD. The prevalence of Afl was significantly higher in biatrial than bicaval anastomosis. Atrial conduction defect, manifested by the increase of terminal force of the P wave in lead V(1) of the surface electrocardiogram, predicted the occurrence of Afl and AF associated with rejection in OHT with a sensitivity of 89% and specificity of 92%. These results demonstrate that the incidence of Afl increased after OHT, which might be a consequence of cellular and humoral rejection, and coronary vasculopathy of the transplanted hearts. PMID- 11472709 TI - Effect of vitamin E on resistance vessel endothelial dysfunction induced by methionine. AB - We tested if vitamin E, a fat-soluble antioxidant, prevents resistance vessel endothelial dysfunction caused by methionine-induced hyperhomocysteinemia in humans. Moderate elevations in plasma homocysteine concentrations are associated with atherosclerosis and hypertension. Homocysteine causes endothelial dysfunction possibly through several mechanisms. No previous study has tested if a fat-soluble antioxidant can prevent endothelial dysfunction caused by experimental hyperhomocysteinemia. Ten healthy subjects participated in a 2 x 2 factorial, double-blind crossover study, receiving L-methionine (100 mg/kg at -6 hours) or vehicle, with and without vitamin E (1,200 IU at -13 hours). Endothelial function of forearm resistance vessels was assessed using forearm blood flow responses to brachial artery administration of endothelium-dependent and endothelium-independent agents. Forearm resistance vessel dilatation to acetylcholine was significantly impaired 7 hours after methionine (placebo, 583 +/- 87% vs methionine 30 +/- 68%; p <0.05). Dilatation to bradykinin was also impaired (placebo, 509 +/- 54% vs methionine 289 +/- 48%; p <0.05). Methionine did not alter vasodilatation to the endothelium-independent vasodilators, nitroprusside, and verapamil. Methionine-induced impairment of resistance vessel dilatation to acetylcholine and bradykinin (p <0.05 vs placebo) was prevented by administration of vitamin E (acetylcholine, p = 0.004; bradykinin, p = 0.004; both vs methionine alone). Experimentally increasing plasma homocysteine concentrations by oral methionine rapidly impairs resistance vessel endothelial function in healthy humans and this effect is reversed with administration of the fat-soluble antioxidant, vitamin E. PMID- 11472710 TI - Pleiotropic effects of statins and their clinical significance. PMID- 11472711 TI - Characteristics of patients with a large discrepancy in coronary artery diameter between quantitative angiography and intravascular ultrasound. PMID- 11472712 TI - Primary stent implantation compared with primary balloon angioplasty for acute myocardial infarction: a meta-analysis of randomized clinical trials. PMID- 11472713 TI - An optimal diagnostic threshold for minimal stent area to predict target lesion revascularization following stent implantation in native coronary lesions. PMID- 11472714 TI - Comparison of six-month outcomes after percutaneous coronary intervention for Whites versus African-Americans. PMID- 11472715 TI - Usefulness of the rescue PT catheter to remove fresh thrombus from coronary arteries and bypass grafts in acute myocardial infarction. PMID- 11472716 TI - Predictors of restenosis following unprotected left main coronary stenting. PMID- 11472717 TI - Frequency of early occlusion and stenosis in bypass grafts after minimally invasive direct coronary arterial bypass surgery. PMID- 11472718 TI - Comparison of operative mortality after coronary artery bypass grafting in Indian subcontinent Asians versus Caucasians. PMID- 11472719 TI - Enhancing recovery in coronary heart disease (ENRICHD): baseline characteristics. PMID- 11472720 TI - Effect of habitual smoking on QT interval duration and dispersion. PMID- 11472721 TI - Restoration of aortopulmonary shunt patency. PMID- 11472722 TI - How many echocardiograms are necessary for follow-up evaluation of patients with Kawasaki disease? PMID- 11472723 TI - Comparison of frequency of patent foramen ovale by transesophageal echocardiography in patients with cerebral ischemic events versus in subjects in the general population. PMID- 11472724 TI - Association of mitral annular calcium with symptomatic peripheral arterial disease in older persons. PMID- 11472725 TI - Usefulness of magnetocardiography for the investigation of fetal arrhythmias. PMID- 11472726 TI - The role of vitamin E in atherogenesis: linking the chemical, biological and clinical aspects of the disease. AB - Atherosclerosis is a disease involving both oxidative modifications and disbalance of the immune system. Vitamin E, an endogenous redox-active component of circulating lipoproteins and (sub)cellular membranes whose levels can be manipulated by supplementation, has been shown to play a role in the initiation and progression of the disease. Recent data reveal that the activities of vitamin E go beyond its redox function. Moreover, it has been shown that vitamin E can exacerbate certain processes associated with atherogenesis. In this essay we review the role of biology of atherosclerosis, and suggest that these two facets decide the clinical manifestation and outcome of the disease. PMID- 11472727 TI - Non-obese diabetic (NOD) mice exhibit an increased cellular immune response to glycated-LDL but are resistant to high fat diet induced atherosclerosis. AB - Diabetes mellitus is one of the major risk factors for atherosclerosis. In recent years several murine models have been developed in an attempt to reproduce the accelerated atherosclerosis by combining induced hyperglycemia with hyperlipidemia. In the present study we wished to examine the effect of spontaneous hyperglycemia and hyperlipidemia induced by high fat diet on atherosclerosis development and on markers of the immune system in diabetes prone NOD mice. We tested two high fat dietary regimens (with or without cholate supplementation) in female NOD mice that either developed or did not develop diabetes. Plasma fasting glucose, lipid profile, antibodies to oxidized-LDL and glycated-LDL were assessed. The spleens from both groups were evaluated for their proliferative response. The extent of atherosclerosis was assessed at the aortic sinus. It was found that the two high fat dietary regimens were insufficient to elicit atherosclerosis in the diabetic and non-diabetic NOD mice. The diabetic hyperlipidemic NOD mice displayed an increased cellular immune response to glycated-LDL in comparison with their non-diabetic littermates. The immune response towards copper oxidized LDL was similar in both groups despite an increased susceptibility of LDL extracted from diabetic hyperlipidemic mice to undergo copper induced oxidation. We conclude that the NOD mouse is highly resistant to atherosclerosis even in the presence of hyperglycemia-hyperlipidemia and increased susceptibility to copper induced LDL oxidation. PMID- 11472728 TI - The fibroproliferative response of arterial smooth muscle cells to balloon catheter injury is associated with increased hyaluronidase production and hyaluronan degradation. AB - Hyaluronan (HA) is a glycosaminoglycan found in greatest amounts in the extra cellular matrix of loose connective tissue. HA has been shown to be closely involved in arterial smooth muscle cell (ASMC) proliferation and migration. No studies have examined the degradation of HA in the vessel wall during proliferation of ASMC. The aim of our study was to determine whether HA degradation was modulated in the injured rat aorta with a catheter balloon. To evaluate HA degradation we quantified the activity of the enzyme which degrades HA (hyaluronidase) and determined HA molecular mass in the aorta. Aorta was analyzed in sham operated aorta (D0) and 14 (D14) days after injury. Intima-media wet weight and DNA content, a parameters reflecting ASMC response to injury, were significantly increased at D14 (+35.5 and +40.8%). HA increased at D14 (+87%) and was mainly expressed in the neointima. Hyaluronidase activity also increased in the aorta at D14 (+25.5%). In the normal aorta, HA was mainly present in a high molecular mass form (2000 kDa). Two low molecular mass HA were also detected (29 and <20 kDa). At D14, the form of 2000 kDa was dramatically increased in comparison to that in normal aorta. In addition, the injured aorta contained a large number of low molecular mass form of HA. To know whether hyaluronidase production in the injured aorta was associated with appearance of new isoforms, we determined the molecular mass of this enzyme. Only one form of hyaluronidase (78 kDa) was present in both groups (D0 and D14). In conclusion, the proliferative response of ASMC to injury in the rat was found to be associated with increased HA degradation. PMID- 11472729 TI - Paraoxonase producing PON1 gene M/L55 polymorphism is related to autopsy-verified artery-wall atherosclerosis. AB - Paraoxonase (PON) is an antioxidative enzyme, which eliminates lipid peroxides. PON has two common polymorphisms (M/L55 and R/Q192) that influence PON concentration and activity. We studied whether the M/L55 or R/Q192 genotype relates with the severity of atherosclerosis of the abdominal aorta, and the mesenteric and common iliac arteries in 123 consecutive autopsy cases (90 males and 33 females, aged 18-93 years). The severity of atherosclerosis in the arteries was evaluated, and the percentage of stenosis was measured. The intimal thickness in the internal elastic lamina (IEL) of the coeliac (CA), superior mesenteric (SMA) and inferior mesenteric (IMA) arteries were measured by light microscopy. The LL homozygous men had more atherosclerotic plaques and complicated lesions in the common iliac arteries (56.8%) than the M allele carriers (28.3%, P=0.007). In logistic regression analysis, age (P<0.001) and the PON M/L55 genotype (P=0.015) were associated significantly with the severity of atherosclerosis in the common iliac arteries independent of smoking status, R/Q192 genotype, hypertension, diabetes mellitus, BMI and sex. The mean intima of the IMA was significantly thicker (P=0.035) and the number of stenotic lesions in SMAs significantly higher (P=0.008) in the LL homozygous men than M allele carriers. In turn, the R/Q192 genotype was not statistically significantly associated with plaque type, intimal thickness in the IEL or with the number of stenotic lesions. This study demonstrates that PON L55 homozygosity is an independent risk factor for autopsy-verified atherosclerosis in Finns. PMID- 11472730 TI - Cross-sectional association of soluble thrombomodulin with mild peripheral artery disease; the ARIC study. Atherosclerosis Risk in Communities. AB - Thrombomodulin, an endothelial membrane glycoprotein, is an essential part of the protein C anti-coagulant pathway. It may also have a role in the regulation of fibrinolysis. We carried out a cross-sectional study to assess the association of soluble thrombomodulin (sTM) with peripheral artery disease (PAD) in a stratified random sample (n=863) of otherwise healthy black and white participants of the Atherosclerosis Risk in Communities (ARIC) Study. PAD was more common in black than in white participants and associated with classical risk factors in an expected manner; positively with age, smoking, hypertension, diabetes (P=0.05), and LDL-cholesterol, and inversely with HDL-cholesterol. Significant positive associations were observed also with fibrinogen and white blood cell count. Overall, the sTM concentration was not a significant predictor of PAD. The association was, however, modified by the level of factor VIII:C in whites (P=0.002 for the interaction), but not in blacks. Protein C was inversely associated with PAD prevalence (odds ratio 0.33, 95% CI 0.18--0.61, P=0.0004). sTM was inversely associated with plasminogen, but no associations with t-PA, PAI 1, or D-dimer were seen. In conclusion, the present results provide some additional evidence on the role of thrombomodulin-protein C pathway in atherosclerotic disease and support our earlier observation on interaction between sTM and factor VIII:C. PMID- 11472731 TI - Analysis of the relationship between triglyceridemia and HDL-phospholipid concentrations: consequences on the efflux capacity of serum in the Fu5AH system. AB - The high triglyceride/low HDL-cholesterol trait is a common finding in the general population. The aim of the present study was to analyze and interpret the relationships between triglycerides (TG), HDL-related parameters and serum cholesterol efflux potential in an asymptomatic population including both normo- and hyperlipidemic individuals. In a large sample (n = 1143) of this population, there was a negative correlation between TG and HDL-cholesterol (HDL-C) (r = 0.49, P<0.0001) whereas the negative correlation between TG and HDL-phospholipid (HDL-PL) (r = -0.29, P<0.0001) was weaker, leading to a strong positive correlation between TG and HDL-PL/C ratio (r = 0.58, P<0.0001). Thus, increased TG concentrations were associated with an enrichment of HDL with PL. Since we have demonstrated previously that HDL-PL is the major determinant for cholesterol efflux potential from Fu5AH rat hepatoma cells, we determined the effect of the variations in HDL lipid composition on the cholesterol efflux capacity in a subsample of 198 subjects. Compared with normolipidemic subjects (NLP) (TG< or = 1.7 mmol/l; LDL-C< or = 4.1 mmol/l, n=58), hypertriglyceridemic subjects (HTG) (TG>1.7 mmol/l, n=63) exhibited lower HDL-C levels (1.08+/-0.21 vs. 1.25+/-0.32, P=0.0003) whereas they showed similar HDL-PL concentrations (1.25+/-0.21 vs. 1.25+/-2.7) and, thus, higher HDL-PL/C ratio (1.17+/-0.15 vs. 1.02+/-0.14, P=0.0001). The relative efflux capacity of serum measured in the Fu5AH system (5% serum, 4 h incubation at 37 degrees C) was on average identical in the HTG and NLP groups. Thus, this study provides evidence that despite decreased HDL concentrations, as determined routinely by the HDL-C assay, some HTG subjects maintained serum cholesterol efflux capacity thanks to the enrichment of HDL with PL. PMID- 11472732 TI - The metabolism of lipoprotein(a) and other apolipoprotein B-containing lipoproteins: a kinetic study in humans. AB - Lipoprotein(a) is a risk factor for cardiovascular disease composed of an apolipoprotein B-containing lipoprotein to which a second protein, apolipoprotein(a), is attached. We investigated in seven subjects with Lp(a) levels of 39--85 mg/dl the metabolism of four apo B-containing lipoproteins (VLDL(1), VLDL(2), IDL and LDL) together with that of apo B and apo(a) isolated from Lp(a). Rates of secretion, catabolism and where appropriate, transfer were determined by intravenous administration of d(3)-leucine, mass spectrometry for measurements of leucine tracer/tracee ratios and kinetic data analysis using multicompartmental metabolic modeling. Apo B in Lp(a) was secreted at a rate of 0.28 (0.17--0.40) mg/kg per day. It was found to originate from two sources -- 53% (43--67) were derived from preformed lipoproteins, i.e. IDL and LDL, the remainder was accounted for by apo B, directly secreted by the liver. The fractional catabolic rates (FCRs) of apo B and of apo(a) prepared from Lp(a) were determined as 0.27 (0.16--0.38) and 0.24 (0.12--0.40) pools per day, respectively, which is less than half of the FCR observed for LDL. Our in vivo data from humans support the view that Lp(a) assembly is an extracellular process and that its two protein components, apo(a) and apo B, are cleared from the circulation at identical rates. PMID- 11472733 TI - Glucosamine enhances platelet-derived growth factor-induced DNA synthesis via phosphatidylinositol 3-kinase pathway in rat aortic smooth muscle cells. AB - Vascular smooth muscle cells play a key role in the development of atherosclerosis. Culture of vascular smooth muscle A10 cells with high glucose for 4 weeks enhanced platelet-derived growth factor (PDGF)-induced BrdU incorporation. Since a long period of high glucose incubation was required for the effect, and it was inhibited by co-incubation with azaserine, the role of hexosamine biosynthesis in the development of atherosclerosis in diabetes was studied in A10 cells. Addition of glucosamine to the culture media enhanced PDGF stimulated BrdU incorporation, and PDGF-induced tyrosine phosphorylation of the PDGF beta-receptor was increased by glucosamine treatment. Of the subsequent intracellular signaling pathways, PDGF-induced PDGF beta-receptor association with PLC gamma was not affected, whereas tyrosine phosphorylation of Shc, subsequent association of Shc with Grb2, and MAP kinase activation were relatively decreased. In contrast, PDGF-induced PDGF beta-receptor association with the p85 regulatory subunit of PI3-kinase and PI3-kinase activation were increased by 20% (P<0.01) and 36% (P<0.01), respectively. The intracellular signaling molecules responsible for the glucosamine effect were further examined using pharmacological inhibitors. Pretreatment with PLC inhibitor (U73122) had negligible effects, and MEK1 inhibitor (PD98059) showed only a slight inhibitory effect on the PDGF-induced BrdU incorporation. In contrast, pretreatment with PI3 kinase inhibitor (LY294002) significantly inhibited glucosamine enhancement of PDGF-induced BrdU incorporation. These findings suggest that glucosamine is involved in the development of atherosclerosis by enhancing PDGF-induced mitogenesis specifically via the PI3-kinase pathway. PMID- 11472734 TI - Effect of calcium channel blockers on proteoglycan synthesis by vascular smooth muscle cells and low density lipoprotein--proteoglycan interaction. AB - Calcium channel blockers are known to retard atherosclerosis. In this study, we tested the hypothesis that one mechanism by which calcium channel blockers retard atherosclerosis is through the modulation of proteoglycan metabolism by vascular smooth muscle cells. We investigated the effect of amlodipine and nifedipine on proteoglycan synthesis by human aortic smooth muscle cells and the ability of the newly synthesized proteoglycans to bind low density lipoprotein (LDL). Confluent smooth muscle cells were incubated with [(35)S]sulfate alone or [(35)S]sulfate and [(3)H]leucine in the presence and absence of different concentrations of amlodipine and nifedipine (0.1--20 microg/ml) for 24 h, and newly synthesized proteoglycans were analyzed. Both amlodipine and nifedipine inhibited proteoglycan synthesis by smooth muscle cells in a dose-dependent manner; however, amlodipine was significantly more potent than nifedipine in this regard. In the presence of 20 microg/ml amlodipine, media and cellular proteoglycans decreased by 56%. This was due to inhibition of de novo proteoglycan synthesis by amlodipine. Compared with the proteoglycans synthesized by control smooth muscle cells, those synthesized by cells exposed to amlodipine were smaller and less sulfated, and contained fewer glycosaminoglycan chains. In addition, proteoglycans synthesized by cells treated with amlodipine bound LDL with low affinity. These results suggest that amlodipine may protect against atherosclerosis through a proteoglycan-mediated mechanism. PMID- 11472735 TI - Heat shock protein (HSP) 47 and collagen are upregulated during neointimal formation in the balloon-injured rat carotid artery. AB - Heat shock protein (HSP) 47, a collagen-specific molecular chaperone, is thought to be essential for the proper processing and secretion of procollagen molecules. We investigated the time course and localization of HSP47 and collagen expression after balloon catheter angioplasty in the rat carotid artery, based on the premise that accumulation of extracellular matrix components is a main feature of intimal hyperplasia in humans and in laboratory animals. Low levels of HSP47 expression were evident in uninjured carotid arteries. Northern blot analysis revealed that HSP47 mRNA expression was markedly stimulated 1--3 days after the induced injury and a high level was maintained for 7 days, followed by a gradual decline for up to 21 days after the injury. These changes in HSP47 expression paralleled changes in alpha 1(I) collagen expression. Immunohistochemical staining revealed colocalization of HSP47 and collagen in smooth muscle cells (SMCs) of the media and intima. In situ hybridization analysis showed that activated SMCs, which proliferated and migrated into the intima, expressed high levels of HSP47. In cultured human aortic SMCs, similar upregulation of HSP47 and alpha1(I) collagen by TGF-beta was noted. These results show that SMCs activated after balloon injury express high levels of HSP47 and collagen during cell proliferation and migration, hence an overproduction of collagen and development of intimal thickening. Thus, HSP47 plays a role in the formation and progression of neointima after angioplasty. PMID- 11472736 TI - Variability of flow mediated dilation: consequences for clinical application. AB - BACKGROUND: Flow mediated vasodilatation (FMD), a non-invasive tool to assess endothelial function, has been shown to have prognostic value for the development of cardiovascular disease. Conventional B-mode ultrasonography has been criticised for its 'limited' resolution in vivo, which complicates reliable detection of the minute diameter changes during reactive hyperaemia. In the present study we evaluated the physical resolution, reproducibility and the capability to detect FMD impairment of a wall tracking system (WTS). METHODS: The resolution of WTS was compared with that of intravascular ultrasound (IVUS) in pig femoral arteries in vivo. Subsequently, intra- and interobserver variability of FMD testing with WTS was evaluated in 75 healthy volunteers. Finally, the effect of smoking as single risk factor for atherosclerosis on FMD in vivo was assessed. RESULTS: WTS and IVUS readings were not different (difference in arterial cross sectional area 1.97 mm(2), r=0.87). Intrasession coefficient of variation in baseline diameter was 1.1% (extremes 0.06--2.0%). Inter-session baseline diameter variation was 3.6 and 3.8% for each observer and 4.1% between observers. Intra-individual variability in FMD between sessions was considerable with coefficients of variation of 13.9% for FMD and 9.3% for NTG. Smokers had impaired FMD responses compared with matched non-smokers (4.7+/-2.4 vs. 9.6+/ 4.4%, P<0.001), whereas NTG induced vasodilatation did not differ (13.4+/-6.2 vs. 15.4+/-5.1%; p=ns). CONCLUSION: WTS is a suitable technique for reproducibly assessing the brachial artery diameter in vivo with a accuracy comparable to that of IVUS. Using this sensitive technique the reproducibility of FMD in vivo proves to be poor mainly due to physiological factors. Whereas this seriously limits the use of FMD as follow-up parameter for individual subjects, FMD is demonstrated to be a useful research tool at group level. PMID- 11472737 TI - Dietary vegetable oil and wood derived plant stanol esters reduce atherosclerotic lesion size and severity in apoE*3-Leiden transgenic mice. AB - The hypolipidemic and anti-atherosclerotic effects of vegetable oil- and wood based dietary plant stanol esters were compared in female apoE*3-Leiden transgenic mice at relevant plasma cholesterol levels. The plant stanol esters derived from vegetable oil (sitostanol 65.7%, campestanol 30.1%) had different contents of sitostanol and campestanol than the plant stanol esters derived from wood (sitostanol 87.6%, campestanol 9.5%) or from a mixture of vegetable oil and wood (sitostanol 73.0%, campestanol 24.7%). The mice (10 per group) received for 38 weeks a control diet or diets containing 1.0% (w/w) plant stanol esters derived from either vegetable oil, wood or a mixture of both. Vegetable oil ( 46%), wood (-42%) and vegetable oil/wood (-51%) plant stanol esters decreased the plasma cholesterol levels (P<0.0001) by reducing the cholesterol content in plasma very low density-, intermediate density- and to a lesser extent in low density-lipoprotein. Plant stanol ester feeding did not change plasma triglyceride levels. Dietary plant stanol esters reduced the atherosclerotic lesion area by 91+/-13% (vegetable oil), 97+/-4% (wood) and 78+/-34% (vegetable oil/wood) (P<0.0001) and the severity from regular intimal fatty streaks/mild plaques (on average type 2--3 lesions) in controls to individual intimal foam cells ( or =90% occlusion in one and > or =40% occlusion in a second coronary artery, while coronary controls had a maximum of 50% occlusion in only one coronary artery. Both males and females, aged 26--64 years were studied. We have previously reported that plasma tHcy is an independent risk factor for coronary atherosclerosis in this study population. In the present analysis, we found that cases had statistically significant higher mean plasma tCys levels than coronary controls and population-based controls (295.8+/-40.2, 279.0+/-35.5 and 282.6+/ 32.4 micromol/l, respectively). The odds ratio (OR) of coronary atherosclerosis for the upper tertile of tCys compared with the bottom tertile was 2.5 (95% confidence interval (CI), 1.4--4.3). However, the association between tCys and coronary atherosclerosis was confounded to a great extent by risk factors (OR, 1.0; 95% CI, 0.5--2.0). The multivariate adjusted OR of coronary atherosclerosis per 1 S.D. increase in plasma tCys was 1.0 (95% CI, 0.8--1.3). The corresponding OR per 1 S.D. increase in plasma tHcy was 1.4 (95% CI, 1.1--1.8). We conclude that plasma tCys, unlike plasma tHcy, is not an independent risk factor for atherosclerosis. PMID- 11472741 TI - Reduced levels of TNF alpha in hypercholesterolemic individuals after treatment with pravastatin for 8 weeks. AB - BACKGROUND: cellular adhesion molecules (CAMs) expressed on the endothelial surface play a key role in the inflammatory process of atherosclerosis, and increased expression of CAMs has been shown in hypercholesterolemic individuals. The expression of CAMs is mediated by several cytokines including tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6). The aim of the present study was to assess the influence of pravastatin 40 mg per day on selected soluble CAMs; intercellular adhesion molecule 1 (ICAM-1), vascular cellular adhesion molecule 1 (VCAM-1), E-selectin, P-selectin and some circulating markers of inflammation; C-reactive protein (CRP) and the cytokines TNF alpha and IL-6. 40 non-diabetic men, age below 70 years, with serum total cholesterol 6--10 mmol/l combined with HDL-cholesterol < or =1.2 mmol/l were included. The study was randomized, double blinded, placebo controlled, cross over designed with 8 weeks intervention periods. Fasting blood samples were drawn after 8 and 16 weeks. RESULTS (MEDIAN VALUES): significant reduction of total cholesterol was achieved after treatment with pravastatin (7.8 on placebo vs. 5.7 mmol/l on pravastatin). TNF alpha was significantly reduced after treatment with pravastatin (1.33 on placebo vs. 1.10 pg/ml on pravastatin, P=0.032), whereas no differences in the levels of the measured sCAMs, CRP and IL-6 were found. Subgroup analysis among smokers versus non-smokers showed a significant reduction in the level of TNF alpha only among the smokers. CONCLUSION: hypercholesterolemic individuals treated with pravastatin 40 mg per day for 8 weeks showed a statistically significant reduction in the levels of TNF alpha as compared with placebo. PMID- 11472742 TI - Increased fibrin monomer plasma concentration in stable coronary artery disease in patients without oral anticoagulation. AB - Blood coagulation has been shown to play a role in the pathogenesis of local thrombus formation in coronary arteries. Increased plasma concentrations of thrombin activation markers such as fibrin monomers (FM) indicate coagulation activation. Therefore, we investigated FM plasma levels in 194 patients (127 nonanticoagulated and 67 anticoagulated) with stable coronary artery disease (CAD) and in 96 healthy controls. FM levels were significantly higher (P<0.0001) in nonanticoagulated patients compared with healthy controls, whereas anticoagulated patients showed significantly lower (P<0.0001) FM levels, respectively. FM levels above 0.50 mg/ml were associated with an odds ratio (OR) of 3.0 (95% confidence interval (CI), 1.7--5.5) for the presence of stable CAD in nonanticoagulated patients. However, the association lost significance after correction for possible confounders such as age, body mass index, total cholesterol, fibrinogen, sex, smoking, arterial hypertension and diabetes mellitus. In conclusion, we found elevated FM plasma levels in nonanticoagulated patients as compared with healthy controls. Elevated FM plasma levels were associated with an OR of 3.0 for the presence of stable CAD in nonanticoagulated patients. PMID- 11472743 TI - Sustained long-term improvement of arterial endothelial function in heterozygous familial hypercholesterolemia patients treated with simvastatin. AB - Patients with heterozygous familial hypercholesterolemia (hFH) are at very high risk for premature coronary heart disease. In the last decade, treatment with statins has reduced cardiovascular mortality in these patients. The aim of this study was to analyze arterial endothelial function assessed as flow-mediated dilatation (FMD) and soluble E-selectin (sE-selectin) levels in patients with hFH under a long-term lipid-lowering treatment. Twenty-five patients who completed the study received a dose of simvastatin to achieve a treatment goal of at least 30% reduction in serum low-density lipoprotein (LDL)-cholesterol (LDL-C) for 52 weeks. Functional and biochemical measurements were taken at entry, and at week 12 and 52 of treatment. FMD was measured by vascular ultrasound of the brachial artery. sE-selectin, tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 were determined by enzyme linked immunosorbent assay (ELISA). LDL-C levels were significantly reduced by treatment at week 12 and maintained at week 52 (reduction vs. baseline, 44+/-12 and 43+/-11%, respectively, P<0.0001). A significant improvement in endothelial function, measured as FMD (baseline, 4.7+/ 6.2%; 12 weeks, 12.3+/-5.9%; 52 weeks, 9.7+/-4.7%; P<0.005) and a reduction in sE selectin levels (baseline, 16.2+/-3.4 ng/ml; 12 weeks, 11.0+/-3.2 ng/ml; 52 weeks, 12.3+/-4.2 ng/ml; P<0.01) were observed. Endothelial-independent relaxation induced by nitroglycerin was not modified during the study. Our results indicate that a long-term treatment with simvastatin produced a sustained beneficial effect in endothelial function in hFH patients. PMID- 11472744 TI - Relationships between alcoholic beverages and cardiovascular risk factor levels in middle-aged men, the PRIME Study. Prospective Epidemiological Study of Myocardial Infarction Study. AB - The relationships between alcoholic beverages and cardiovascular risk factors were assessed in 6730 men living in France or Northern Ireland. In France, all alcoholic beverages were significantly correlated with body mass index (BMI), systolic (SBP) and diastolic blood pressure (DBP), high density lipoprotein (HDL) parameters, PAI-1 and Factor VII, whereas only wine was negatively related with fibrinogen levels. After adjusting for center, age, BMI, educational level, smoking and marital status, wine had a lesser effect on blood pressure, triglyceride, apo B and LpE:B levels than beer. Wine was associated with lower fibrinogen levels and beer with higher PAI-1 activity levels independent of the amount of alcohol consumed. In Northern Ireland, wine was negatively correlated with BMI, triglycerides, LpE:B and fibrinogen, whereas beer was positively correlated with SBP and DBP, triglycerides, HDL, apoprotein A-I and fibrinogen. Multivariate analysis showed wine to be positively associated with HDL parameters, and negatively with fibrinogen levels. Wine was also associated with higher LpA-I levels and lower fibrinogen levels independent of the amount of alcohol consumed. We conclude that alcohol consumption is related to lipid, lipoprotein and haemostatic variables, but the magnitude of the relationships depends on the type of alcoholic beverage. Also, some effects might be related to non-alcoholic components. PMID- 11472745 TI - High plasma homocyst(e)ine levels in elderly Japanese patients are associated with increased cardiovascular disease risk independently from markers of coagulation activation and endothelial cell damage. AB - Elevated plasma homocyst(e)ine is a risk factor for cardiovascular disease (CVD) in many populations, but the relationship between homocyst(e)ine and CVD in Japanese subjects has been unclear. It has been hypothesized that the link between homocyst(e)ine and CVD may be mediated in part by activation of coagulation and endothelial cell injury in the elderly Japanese subjects. To further evaluate this hypothesis, the present cross-sectional study was designed to assess the relationships among plasma homocyst(e)ine concentrations, risk of CVD, and markers of coagulation (fibrinogen, FVII, F1+2, FVIIa and FXIIa) and endothelial cell damage (vWF and thrombomodulin) in 146 elderly Japanese subjects (79 healthy controls and 67 patients with CVD). The geometric mean (range) of plasma homocyst(e)ine concentrations was 10.2 (3.2--33) micromol/l in 79 Japanese healthy elderly subjects. As expected, healthy female and male elderly subjects had homocyst(e)ine levels that were 2.5 and 5.3 micromol/; higher, respectively, compared to healthy young control subjects (n=62). Healthy young and elderly men had homocyst(e)ine levels that were 1.7 and 4.5 micromol/l higher, respectively, compared to values in women. This higher plasma homocyst(e)ine levels in the elderly subjects were negatively correlated with levels of folic acid, albumin and total cholesterol, but were not significantly related to markers of coagulation or endothelial cell-damage. The results of multiple logistic regression analyses suggested that high homocyst(e)ine levels were independently related to CVD risk. In addition, levels of FVIIa, and F1+2 were significantly higher in elderly Japanese patients with CVD compared to elderly subjects without CVD, but were unrelated to plasma homocyst(e)ine concentrations. In summary, elevated plasma concentrations of homocyst(e)ine, FVIIa, and F1+2 were associated with increased risk of CVD in elderly male and female Japanese subjects, but the association between homocyst(e)ine and CVD was unrelated to abnormalities in markers of coagulation and endothelial cell damage in this population. PMID- 11472746 TI - The methionine synthase reductase (MTRR) A66G polymorphism is a novel genetic determinant of plasma homocysteine concentrations. AB - Epidemiological evidence has revealed that an elevated plasma homocysteine level (hyperhomocysteinemia) confers an increased risk of cardiovascular disease and neural tube defects. Hyperhomocysteinemia is caused by both nutritional (e.g. folate, vitamins B(6) and B(12)) and genetic factors, including functional polymorphisms of key enzymes involved in homocysteine metabolism. One such enzyme, methionine synthase reductase (MTRR), maintains adequate levels of methylcob(III)alamin, the activated cofactor for methionine synthase, which catalyzes the remethylation of homocysteine to methionine. A common MTRR polymorphism, i.e. a 66 A-->G substitution specifying an isoleucine to methionine substitution (I22M), was recently identified. To assess the influence of this polymorphism on total plasma homocysteine (tHcy), we undertook a genotype/phenotype analysis in a study population of 601 Northern-Irish men, aged 30--49, for which biochemical and genetic data relevant to folate/homocysteine metabolism had already been acquired. The 66AA genotype has a frequency of 29% in this population. We established that there was a significant influence of MTRR genotype on tHcy ranking (P=0.004) and that the 66AA genotype contributes to a moderate increase in tHcy levels across the distribution [OR 1.59 (95% CI: 1.10- 2.25) for the 66AA genotype to be in the upper half of the tHcy distribution, P=0.03]. The homocysteine-elevating effect of the 66AA genotype is independent of serum folate, vitamin B(12) and vitamin B(6) levels. Based on published estimates of the enhanced cardiovascular disease risk conferred by defined increments of plasma tHcy, we estimate that 66AA homozygotes have, on average, an approximately 4% increase in cardiovascular disease risk compared to 66GG homozygotes. This study provides the first evidence that the MTRR A66G polymorphism significantly influences the circulating tHcy concentration. PMID- 11472747 TI - Further evidence against the implication of active cytomegalovirus infection in vascular atherosclerotic diseases. AB - The possible contribution of cytomegalovirus (CMV) to pathogenetic events associated with atherosclerotic lesion establishment and progression is still controversial. We evaluated the possibility that active ongoing CMV infection could be correlated to evolution of unstable atheromatous lesion, by analyzing patients suffering from unstable angina (n=61), acute myocardial infarction (n=43), stable angina (n=26) and peripheral arteriopathy (n=22) as compared to healthy subjects (n=30). Particularly, we assessed: past exposure to CMV by evaluating anti-CMV IgG antibodies; ongoing CMV infection by evaluating anti-CMV IgM antibodies and circulating interleukin (IL)-8 in serum; and CMV DNAemia in peripheral blood mononuclear cells (PBMC). Mean IgG values were significantly increased in patients from all groups, as compared to healthy subjects. CMV specific IgM, as well as CMV DNAemia, were undetectable in both controls and patients. Circulating IL-8, significantly elevated in a group of individuals experiencing active CMV infection, was not significantly higher in cardiovascular disease patients, as compared to control subjects. These findings confirm previous evidence from the increased exposure to CMV infection in patients with atheromatous lesions. However, they provide further evidence against a direct implication of active systemic CMV infection in the pathogenesis of cardiovascular diseases, particularly those involving plaque instability. PMID- 11472748 TI - Heart rate variability after long-term treatment with atorvastatin in hypercholesterolaemic patients with or without coronary artery disease. AB - Low heart rate variability (HRV) level, indicative of impaired autonomic function, is associated with an increased risk of cardiovascular morbidity and mortality and is negatively affected by hypercholesterolaemia. In order to test the hypothesis that significant low density lipoprotein (LDL) cholesterol reduction after treatment with a statin will have a beneficial effect on HRV level in hypercholesterolaemic patients with or without coronary artery disease (CAD), forty consecutive patients (28 men and 12 women) with a median age of 61, range (17--70) years were studied. Twenty had stable CAD and 20 were free of CAD at baseline. Twenty healthy volunteers, of similar age and gender as the patients, were used as controls. Patients were treated with atorvastatin (20 mg/day) for 2 years. Changes in lipid parameters and HRV indices were assessed at baseline and 2 years later in all subjects. In both patient subgroups a significant beneficial change in all lipid parameters (more pronounced in the CAD+ subgroup) and a significant beneficial modification in HRV time and frequency domain indices was recorded (more pronounced in the CAD- subgroup), while lipid parameters and HRV indices remained unchanged in the control group. A correlation between LDL concentrations and most of the HRV indices was found at baseline in both patient subgroups, while no such correlation was found between values or their percent changes after hypolipidaemic treatment. These data suggest that treatment with atorvastatin improves autonomic function, as reflected by an increase in HRV level, and this may be a likely mechanism, at least in part, for the reduction in clinical events reported by the landmark survival studies with statins in primary and secondary CAD prevention. Perhaps, if this finding is confirmed by larger studies, HRV level may prove to be a useful tool for risk-stratification and treatment guide in high-risk patients with hypercholesterolaemia, regardless of CAD. PMID- 11472749 TI - The relation of oxidized LDL autoantibodies and long-term hormone replacement therapy to ultrasonographically assessed atherosclerotic plaque quantity and severity in postmenopausal women. AB - BACKGROUND: In epidemiologic studies, the incidence of atherosclerosis rises soon after menopause in women, and hormone replacement therapy (HRT) has proved to be useful in preventing onset of clinical manifestations of the disease. However, it is not known how HRT affects sonographically determined atherosclerotic severity (AS) and number of atherosclerotic plaques (NAP) in large arteries. Furthermore, it is not clear how HRT affects oxidation of low density lipoproteins (LDL), which obviously has an important role in the pathogenesis of atherosclerosis. OBJECTIVES: The purpose of the study was to determine whether HRT has a beneficial effect on sonographically determined AS and NAP in large arteries of 101 postmenopausal women compared to 40 controls without HRT. We also studied the interaction of HRT and antibodies against oxidized LDL on AS and NAP progression. RESULTS: Estradiol valerate alone, combined estradiol valerate-levonorgestrel and combined estradiol valerate-medroxyprogesterone acetate therapy are each associated with lower NAP and AS as compared to controls without HRT. In a multiple regression model explaining NAP in the whole study population, the strongest predictors were HRT (P=0.0006) and copper-oxidized LDL cholesterol autoantibodies (P=0.0491). DISCUSSION: Our findings indicate that postmenopausal HRT is associated with a lower total number of atherosclerotic plaques and less severe atherosclerotic lesions, as compared to controls without HRT, and that this outcome may be associated with the effect of HRT on LDL cholesterol oxidation. PMID- 11472750 TI - Excess coronary heart disease in Familial Combined Hyperlipidemia, in relation to genetic factors and central obesity. AB - AIM: To determine the prevalence of non-fatal coronary artery disease (CAD) in kindred with Familial Combined Hyperlipidemia (FCHL) in relation to various cardiovascular risk factors and DNA variation in the apo AI-CIII-AIV gene cluster. METHODS AND RESULTS: Data were collected from 18 Dutch FCHL probands, 202 living first and second degree relatives, and 175 spouses. Probands and first degree relatives showed dyslipidemia, increased plasma insulin and glucose concentrations, higher waist--hip ratio (WHR), and blood pressure, than spouses. The frequency of the minor alleles M2 and S2 was increased in probands and first degree relatives. The Odds Ratio for CAD was 5.3 in male FCHL relatives (P=0.005), and 5.1 in all FCHL relatives (P=0.001). First and second degree relatives had a markedly reduced CAD-free life-span (logrank vs. spouses: P<0.001 and P=0.03, respectively). The presence of the S2, but not M2, minor allele, showed a marked reduction in CAD-free life-span (logrank S2 present vs. S2 absent: P=0.035). CONCLUSION: Men with FCHL have a severely increased risk of CAD, that appears to be mediated through genetic relation to the proband as the strongest independent risk factor for CAD, followed by increased WHR. PMID- 11472751 TI - C-reactive protein in patients with familial hypercholesterolemia: no effect of simvastatin therapy. AB - Patients with familial hypercholesterolemia (FH) are especially at risk for premature cardiovascular disease (CVD). Recent studies revealed C-reactive protein (CRP) as a strong predictor of future first or recurrent CVD events, suggesting that CRP plays an important role in the development of atherosclerosis. The aim of this study was to evaluate the effect of one year of simvastatin treatment on serum levels of CRP and to assess the influence of risk factors for CVD on CRP concentrations in patients with FH. We measured baseline CRP levels in 337 patients with FH. A second blood sample, collected after one year of treatment with simvastatin (20--40 mg once daily) was measured in a subgroup of 129 patients. Patients with CVD present at baseline had significantly higher serum levels of CRP (2.26 mg/l versus 1.55 mg/l, P<0.001). CRP levels were associated with smoking, body mass index, age, levels of triglycerides (TG), and the use of NSAIDs or anticoagulation drugs. Simvastatin therapy significantly improved lipid profiles in the intervention group. There was a small, but non significant decrease of CRP levels upon treatment. CRP decreased from 1.51 mg/l median (interquartile range (IQR) 0.76--3.41) at baseline to 1.24 mg/l median (IQR 0.72--2.92) after treatment, (P=0.328). In conclusion, CRP levels were associated with the presence of CVD in FH patients. Simvastatin therapy had no significant effect on CRP levels in these patients. PMID- 11472752 TI - Plasma insulin, leptin, and soluble TNF receptors levels in relation to obesity related atherogenic and thrombogenic cardiovascular disease risk factors among men. AB - Obesity is related to cardiovascular disease (CVD) morbidity and mortality, however, the mechanisms for the development of obesity-induced CVD risk remain unclear. Hyperinsulinemia and insulin resistance are considered key components in the metabolic cardiovascular syndrome and as independent risk factors for CVD. Plasma leptin and tumor necrosis factor-alpha (TNF-alpha), two adipocyte products, are also proposed to be associated with the development of CVD risk. The purpose of this study is to evaluate the association of plasma leptin, soluble TNF receptors (sTNF-R), and insulin levels as possible mediators of the effect of obesity on atherogenic and thrombogenic CVD risk factors among men. From the Health Professionals Follow-up Study (HPFS), we selected 268 men, aged 47--83 years, who were free of CVD, diabetes, and cancer (except non-melanoma skin cancer), and who had provided a fasting blood sample in 1994. We measured plasma insulin and leptin levels by radioimmunoassay and sTNF-R levels by ELISA. Men in the highest quintile of body mass index (BMI, mean=30.5 kg/m(2)) were less physically active and had a more adverse cardiovascular lipid and homeostatic profile, as indicated by levels of insulin, triglyceride (TG), tissue plasminogen activator (t-PA) antigen levels, and apolipoprotein A1 (Apo-A1). In a multivariate regression model controlling for age, smoking, alcohol intake, physical activity and diet, BMI was inversely associated with HDL-cholesterol (HDL-C) and Apo-A1 and positively associated with TG, Apo-B and t-PA antigen levels. The associations between BMI and these CVD risk factors were only slightly changed after adjusting for leptin and/or sTNF-R; but were substantially attenuated after controlling for insulin levels. These data suggest that the association between obesity and biological predictors of CVD may be mediated through changes in plasma insulin, rather than leptin or sTNF-R levels. However, plasma leptin may still play a role in CVD through independent effects on lipid metabolism. PMID- 11472753 TI - Effect of simvastatin on monocyte adhesion molecule expression in patients with hypercholesterolemia. AB - Increased monocyte adherence to the vessel wall is one of the earliest events in atherosclerosis. The mechanism by which hypercholesterolemia causes alterations in endothelial adhesiveness for monocytes is unclear. This study sought to determine if monocyte adhesion molecule expression is affected by low-density lipoprotein (LDL)-cholesterol levels. Patients with hypercholesterolemia and stable coronary artery disease were compared with those without major cardiovascular risk (control). Patients with hypercholesterolemia were treated with simvastatin 20--40 mg/day for 8--10 weeks. Blood samples were examined with flow cytometry assays at baseline and after cholesterol-lowering therapy. Monocyte CD11b and CD14 adhesion molecule expression, measured as fluorescence intensity, were significantly (P<0.0001) higher in hypercholesterolemic patients before the study (176.9+/-9.8 and 138.0+/-4.8, respectively) when compared with that in control subjects (97.2+/-8.1 and 84.0+/-6.4, respectively). Both decreased markedly with treatment: to 118.8+/-6.9 and 103.1+/-3.9, respectively. Monocyte L-selectin expression was significantly lower in patients with hypercholesterolemia before treatment (43.0+/-3.0) when compared with control subjects (79.9+/-2.7), and it increased markedly with treatment (54.2+/-2.5). LDL levels correlated directly with both CD11b and CD14 expression and correlated inversely with L-selectin expression. These data show that hypercholesterolemia affects monocyte adhesion molecule expression which, in turn, decreases with statin-induced plasmatic cholesterol reduction. Such perturbations in monocyte function likely represent a proinflammatory response to hypercholesterolemia and may have a role in the early progression of atherogenesis. PMID- 11472754 TI - The use of Achilles tendon ultrasonography for the diagnosis of familial hypercholesterolemia. AB - Differentiating FH from other causes of hypercholesterolemia has important clinical and therapeutic implications but is often not possible by standard clinical criteria. As accumulation of cholesterol in tendon is generally considered as pathognomonic of FH, we evaluated the sensitivity and specificity of clinical and ultrasonographic tendon characteristics using the data of 127 genetically ascertained FH and 160 controls with various lipid profiles. Upon clinical examination, none of the controls and 29% of FH individuals (17% FH women and 38% FH men) presented with xanthomata in Achilles tendons, but no female and only 6% of male FH patients also showed xanthomata in the extensor tendon of the hand. Amongst all possible quantitative parameters (thickness, breadth, section and roundness) of Achilles tendon (AT) measured by ultrasonography, the thickness presented the best receiver operating curves. AT thickness above 5.8 mm was the most useful threshold for diagnosis of FH, procuring sensitivity of 75% and specificity of 85%. Analysis of variation of AT thickness with age and sex indicated that this clinical criterion performed better in females older than 45 and in males under 45. In patients carrying the APOB-R3500Q mutation, AT-thickness appeared significantly less important compared with those carrying LDLR mutations. In conclusion, this study recommends identification of possible FH individuals amongst hypercholesterolemic patients using a criteria of AT-thickness over 5.8 mm eventually associated with a specific genetic test for APOB-R3500Q mutation. PMID- 11472755 TI - Chylomicron remnant metabolism in familial hypercholesterolaemia studied with a stable isotope breath test. AB - Chylomicron remnant metabolism was studied using a stable isotope breath test in 25 patients with familial hypercholesterolaemia (FH) (10 homozygotes, 15 heterozygotes), and in 15 normolipidaemic controls. A lipid emulsion mimicking the composition of chylomicron remnants and labelled with cholesteryl (13)C oleate was injected intravenously; (13)CO(2) was measured subsequently in breath using isotope-ratio mass spectrometry. The fractional catabolic rate (pools/h) of the emulsion, derived from a compartmental model, did not differ significantly among the groups: homozygous FH mean 0.20 (S.E.M. 0.05), heterozygous FH 0.12 (0.02), controls 0.16 (0.03). We suggest that the catabolism of chylomicron remnants from plasma is not impaired in FH and that the hepatic uptake of these particles is not dependent on functional LDL receptors. PMID- 11472756 TI - A novel splice-site mutation in intron 7 causes more severe hypercholesterolemia than a combined FH-FDB defect. PMID- 11472757 TI - Natural oviposition-deterring chemicals in female cotton bollworm, Helicoverpa armigera (Hubner). AB - Observation of the oviposition behavior of cotton bollworm demonstrated the existence of substances that can act as oviposition deterrents. The scales, abdomens and tarsi of the moths, the most likely carrier, were extracted for the substances. An acetone extract exhibiting an oviposition deterrent effect was analyzed using gas chromatography and combined gas chromatography-mass spectroscopy. Three components, 4-methyl-4-hydroxyl-pentanone-2, hexadecanoic acid and (Z)-9-octadecenoic acid, were identified, being present in the ratio 1:2:2. Experiments with synthetic mixtures indicated that a mixture of equal amounts of the two organic acids exhibited similar repellent and deterrent activity to a synthetic mixture of all three components. It is concluded that the natural oviposition-deterring pheromone is a mixture of hexadecanoic acid and (Z) 9-octadecenoic acid. PMID- 11472758 TI - Mating system evolution in sperm-heteromorphic Drosophila. AB - In Drosophila species of the obscura group, males exhibit sperm-heteromorphism, simultaneously producing both long sperm, capable of fertilization, and short sperm that are not. The production of multiple sperm types calls into question whether mating system correlates, such as sperm length and number trade-offs and female remating behavior, are the same as previously described in sperm monomorphic systems. We examine three obscura group species, D. pseudoobscura, D. persimilis, and D. affinis that differ significantly in the lengths of their long fertilizing sperm, to test predictions about the relationship between sperm length and four mating system characters: male age at sexual maturity; sperm number; female remating; and male reproductive output. In D. affinis, where males produce the longest fertilizing sperm, their sexual maturity is delayed and they produce fewer long sperm compared to the other two species, as predicted if long sperm are costly to produce. Female D. affinis, although they receive fewer sperm than females of the other two species, do not remate more frequently or produce fewer progeny from a single mating. Different responses between sperm heteromorphic and sperm-monomorphic systems underscore the complex nature of the coevolution between male and female mating system characters. PMID- 11472759 TI - Surface characteristics of foreign targets that elicit an encapsulation response by the moth Pseudoplusia includens. AB - Hemocytes from the moth Pseudoplusia includens encapsulate a variety of biotic and abiotic targets. Prior studies indicated that granular cells are usually the first hemocyte type to attach to foreign targets. Thereafter, large numbers of plasmatocytes attach to the target and form a capsule. To identify surface features that induce an encapsulation response, chromatography beads that differed in matrix composition, charge, and functional groups were tested using in vitro and in vivo bioassays. We first conducted in vitro assays using hemocytes with no plasma components present. These experiments indicated that bead types having sulfonic, diethylaminoethyl, and quaternary amine functional groups were encapsulated significantly more often than beads with other functional groups. Charge also significantly affected encapsulation with positively charged beads being encapsulated more often than negatively charged or neutral beads. In vitro assays using purified populations of hemocytes confirmed that these targets were recognized as foreign by granular cells, and that plasmatocytes only formed capsules after granular cells attached to the target. Bead types that were encapsulated under these in vitro conditions were always rapidly encapsulated when injected into P. includens larvae. However, some bead types, like CM-Sephadex, not encapsulated in vitro were encapsulated in vivo if left in the insect hemocoel for a longer period of time (ca. 24 h). Purified plasmatocytes encapsulated these beads in vitro if they were preincubated in plasma. Basic characterization studies suggest these humoral recognition molecules are proteins or small peptides. Comparative studies with other species of noctuid moths also indicated that encapsulation of some bead types differed significantly among species. Collectively, these results reveal that P. includens recognizes some targets as foreign by pattern recognition receptors on granular cells, whereas others are recognized by pattern recognition molecules in plasma. The binding affinities of these recognition molecules also appear to differ among closely related species of Lepidoptera. PMID- 11472760 TI - Effects of age, diet, female density, and the host resource on egg load in Anastrepha ludens and Anastrepha obliqua (Diptera: Tephritidae). AB - Oocyte counts, used as a measure of egg load, were compared among three different age groups (15, 30 and 45 days) of two polyphagous species of tephritid fruit flies, Anastrepha ludens and Anastrepha obliqua, which were exposed to varying conditions of diet (sucrose vs sucrose and protein), availability of oviposition substrate (present vs absent), adult female density (1, 2 and 4 females/cage), and semiochemical context (presence vs absence of male pheromones and fruit volatiles). In both species, oocyte counts were higher in older females and for females fed sucrose and protein than for females fed sucrose only. The presence of artificial oviposition substrates influenced oocyte counts in A. obliqua, but not in A. ludens. Female density influenced oocyte counts in both species. Females maintained in groups had higher egg loads than isolated females. Finally, preliminary evidence suggests that semiochemical context influenced oocyte counts. Counts were highest for females in a room containing both fruit volatiles and male pheromone, lowest for females in a room containing neither volatiles nor pheromone, and intermediate for females in rooms containing either volatiles or pheromone but not both. Our results suggest that egg load is influenced by environmental factors in different ways in these two species. Egg load in A. obliqua, a species whose host fruits are highly ephemeral, is responsive to access to the host resource. By contrast, in A. ludens, a species infesting less ephemeral fruit, female density and age played a more important role than host stimuli. The role of ovarian maturation and oviposition in mediating these effects, as well as implications for mass rearing and pest management, are discussed. PMID- 11472761 TI - Source of the host marking pheromone in the egg parasitoid Trissolcus basalis (Hymenoptera: Scelionidae). AB - After oviposition, Trissolcus basalis females always mark the host's surface, depositing host marking substances for herself and to warn other ovipositing females. The perception of these host marking substances, probably through the antennae, can induce the female to leave and seek healthy hosts. Parasitoid females exposed to conspecific parasitized egg masses left the host egg masses significantly more often than when exposed to non-parasitized egg masses. More egg mass leaving behavior also was observed when the egg masses were treated with Dufour's gland secretion but not when treated with secretion from the common oviducts. The common oviduct has a secretory epithelium that produces electron dense vesicles, probably containing proteinaceous substances. The secretory cells of the accessory gland, Dufour's gland, contain electron-lucid vesicles, whose secretion appears to be a lipid similarly to that found in pheromone secreting glands. Ultrastructural and behavioral evidence suggests that Dufour's gland is the host marking pheromone source. PMID- 11472762 TI - The extracellular matrix protein lacunin is expressed by a subset of hemocytes involved in basal lamina morphogenesis. AB - The extracellular matrix protein of Manduca sexta known as lacunin is localized to basal laminae and granular cells of the hemolymph. Circulating granular cells increase in both number and size as pupal basal laminae break down after the initiation of adult development. Basal laminae of wing epithelia break down as ecdysteroid levels rise during adult development, and lacunin immunoreactivity concurrently passes from basal laminae to endocytic vacuoles of granular cells. Granular cells not only endocytose lacunin protein that they salvage from the remnants of the old basal laminae but they also express transcripts for lacunin. As new adult basal laminae form several days later, the number of circulating granular cells decreases as lacunin immunoreactivity appears in the new basal laminae. PMID- 11472763 TI - Resistance to stress as a function of age in transgenic Drosophila melanogaster overexpressing Hsp70. AB - This study investigated the resistance to stress as a function of age in Drosophila melanogaster overexpressing Hsp70. The resistances to starvation, paraquat, and cold in flies from 1 to 7 week-old have been measured. The line carrying the insertion vector without the transgenes is more resistant to starvation and cold than the parental and transgenic lines. In contrast, transgenic flies carrying extra-copies of hsp70 are more resistant to paraquat, however this is due to an especially high resistance in two age groups compared to all the other groups. I showed that exposure to a mild heat shock does not increase starvation resistance, slightly increases paraquat resistance, and strongly increases cold resistance. The transgenic flies expressing Hsp70 at higher levels after the heat shock do not exhibit enhanced stress resistance compared to control lines expressing less Hsp70 after the heat shock. The lack of effect of a mild heat shock on starvation and paraquat resistance is not due to a disappearance of the effect with age, since no effect is observed at any age. In contrast, when an effect of Hsp70 induction is observed as on cold resistance, this effect is still observed in old flies. PMID- 11472764 TI - Nestmate recognition in Parischnogaster striatula (Hymenoptera Stenogastrinae), visual and olfactory recognition cues. AB - The recognition of nestmates from alien individuals is a well known phenomenon in social insects. In the stenogastrine wasp Parischnogaster striatula, we investigated the ability of females to recognize nestmates and the cues on which such recognition is based. Recognition of nestmates was observed in naturally occurring interactions between wasps approaching a nest and the resident females on that nest. This recognition was confirmed in experiments in which nestmates or alien conspecifics were presented to resident females. In naturally occurring interactions, nestmates generally approach their nest with a direct flight, while aliens usually hover in front of the nest before landing. In experiments in which the presented wasps were placed close to the nest in a direct manner, antennation of the presented wasp generally occurred, indicating that chemical cues are involved. Experiments in which dead alien individuals, previously washed in hexane, and then reapplied with extracts were recognized by colonies giving further evidence that chemical cues mediate nestmate recognition. Epicuticular lipids, known to be nestmate recognition cues in social insects, were chemically analysed by GC-MS for 44 P. striatula females from two different populations (13 different colonies). Discriminant analysis was performed on the data for the lipid mixture composition. The discriminant model showed that, in the samples from these two populations, 68.2% and 81.9% of the specimens could be correctly assigned to their colony. PMID- 11472765 TI - Enhanced drought tolerance of a soil-dwelling springtail by pre-acclimation to a mild drought stress. AB - The springtail Folsomia candida has a highly permeable cuticle, but is able to survive several weeks at 98.2%RH. This corresponds to a water potential deficit of about 17bars between the environment and the normal osmotic pressure of the body fluids of this animal. Recent studies have shown a water vapour absorption mechanism by accumulation of sugars and polyols (SP) in F. candida, which explains how this species can survive dehydrating conditions. In the present study, adult F. candida were pre-acclimated at 98.2%RH to induce the accumulation of SP, and were subsequently exposed for additional desiccating conditions from 98 to 94%RH. Activity level, water content, osmotic pressure of body fluids and SP composition were investigated. After the desiccation period, the animals were rehydrated at 100%RH and survival was assessed. The results showed that F. candida survived a more severe drought stress when it had been pre-acclimated to 98.2%RH before exposure to lower humidity. This species was able to maintain hyperosmosity to the surroundings at 95.5%RH, suggesting that it can absorb water vapour down to this limit. Below this limit, trehalose levels increased while myo inositol levels decreased. We propose that this is a change of survival strategy where F. candida at mild desiccation levels seek to retain water by colligative means (remain hyperosmotic), but at severe desiccation levels switches to an anhydrobiotic strategy. PMID- 11472766 TI - Taste cell responses in the polyphagous arctiid, Grammia geneura: towards a general pattern for caterpillars. AB - The caterpillars of Grammia geneura are polyphagous as individuals. Electrophysiological responses of its medial and lateral galeal styloconic sensilla to 21 amino acids, 6 carbohydrates, 10 chemically diverse plant secondary compounds and two inorganic salts were examined. In the medial sensillum, a single cell responded to 8 amino acids, 3 carbohydrates, and the iridoid, catalpol, which is present in a favored hostplant. In the lateral sensillum, one cell responded to amino acids and another to fructose. Two cells in each sensillum responded to secondary compounds and it is suggested that the same cells are stimulated by inorganic salts. There was no evidence of a separate salt-sensitive cell. Phenylalanine stimulated a deterrent cell in the medial sensillum and was behaviorally deterrent. Some essential amino acids did not stimulate any cells and it is suggested that a small number of amino acids (sometimes non-essential) may serve as indicators of nutrient quality. Sugars probably serve as the primary phagostimulants because they are in relatively high concentrations in plants. It is proposed that taste receptor cells should be categorized primarily by their behavioral effects as phagostimulatory or deterrent, rather than their specific ranges of responsiveness. This would emphasize the basic similarities across taxa. PMID- 11472767 TI - Hormonal control of male horn length dimorphism in Onthophagus taurus (Coleoptera: Scarabaeidae): a second critical period of sensitivity to juvenile hormone. AB - Male dung beetles (Onthophagus taurus) facultatively produce a pair of horns that extend from the base of the head: males larger than a threshold body size develop long horns, whereas males that do not achieve this size develop only rudimentary horns or no horns at all. Using topical applications of methoprene, we identified a sensitive period during the feeding stage of third (final) instar larvae when application of methoprene shifted the threshold body size for horn expression. Male larvae that received methoprene at this time delayed horn production until they attained a larger threshold body size than acetone-treated control larvae. This new sensitive period occurs earlier than a sensitive period previously reported for male horn regulation, and it coincides with a morph-specific pulse of ecdysteroid secretion described for this species. It appears that male horn expression is influenced by endocrine events at two different periods of larval development. We incorporate these results into an expanded model for the endocrine regulation of male horn expression. PMID- 11472768 TI - How female caterpillars accumulate their nutrient reserves. AB - Female Lepidoptera are often heavier than males. We examined the importance of consumption and post-ingestive processing as mechanisms for female Heliothis virescens larvae to meet the protein and carbohydrate requirements. In experiments in which caterpillars had a choice of diets, enabling them to select an appropriate protein and carbohydrate intake, females caterpillars ate more carbohydrate than males, but only on the heavily carbohydrate biased treatment. Overall, the sexes were not distinguished according to the selective feeding behavior, but females accumulated more protein and carbohydrate over the whole instar than the males did. Additionally, when given no choice, females ate more than males and accumulated more protein provided the diet contained a high proportion of protein. If they were reared on a high carbohydrate diet, there were no differences between the sexes. Our results indicate that female H. virescens larvae accumulate protein by regulating both intake and post-ingestive processing on high protein foods. In the field, late instar H. virescens feed on anthers, which are protein-rich and have the highest amino acid content relative to other cotton floral tissues. PMID- 11472769 TI - Olfactory receptor neurons detecting plant odours and male volatiles in Anomala cuprea beetles (Coleoptera: Scarabaeidae). AB - We have identified several types of olfactory receptor neurons in male and female Anomala cuprea beetles. The receptor neurons were sensitive to female sex pheromone components, flower volatiles, green leaf volatiles and unknown volatiles from males. Olfactory sensilla were located on three lamellae forming the antennal club. There was a clear spatial separation between some types of sensilla on each lamella. Receptor neurons for the two sex pheromone components were situated in sensilla placodea covering a specific area on each lamella in both males and females. All sex pheromone receptor neurons were found in these sensilla. Most other receptor neurons were located in a longitudinal, heterogeneous streak formed by various types of sensilla. Receptor neurons for plant-derived compounds appeared to be specialists with a high sensitivity to their respective key compound. The most remarkable among these are the green leaf volatile-specific receptor neurons, which were both sensitive and selective, with the key compound being at least 1000 times as effective as any other compound. These green leaf volatile detectors are apparently homologous to detectors recently found in the scarab Phyllopertha diversa. Our results emphasize the role of single-sensillum recordings as a tool in the identification of biologically active odours. PMID- 11472770 TI - A comparison of nectar- and honeydew sugars with respect to their utilization by the hymenopteran parasitoid Cotesia glomerata. AB - Fourteen naturally occurring sugars were individually tested with respect to their effect on Cotesia glomerata longevity. Parasitoids kept with solutions of either sucrose, glucose and fructose lived for >30 days. This constitutes a factor 15 increase in life span in comparison to control individuals kept with water only. Stachyose, mannose, melezitose, melibiose, maltose and erlose increased parasitoid longevity by a factor of 11.2-6.9. Solutions of galactose and trehalose had a marginal, but still significant effect. Lactose and raffinose did not raise parasitoid longevity, while rhamnose actually reduced parasitoid survival. In an additional experiment, the relationship between quantity of sugar consumption and longevity was established for all 14 sugars. To study the effect of an unsuitable sugar in sugar mixtures, a range of glucose:rhamnose mixtures was tested. Even at 20% of the sugar mixture rhamnose suppressed the nutritional benefit of the 80% glucose. The nutritional suitability of the sugars shows a positive correlation with the previously reported gustatory response towards the individual sugars. Patterns of sugar utilization are discussed with respect to hydrolytic enzymes and carbohydrate biochemical characteristics. Our findings for C. glomerata are compared to patterns of sugar utilization reported for other species. The comparison between C. glomerata and its host Pieris brassicae reveals that the parasitoid is capable of utilizing a range of sugars that are unsuitable to its herbivorous host. This specificity opens up opportunities to select food supplements for biological control programs that selectively target the antagonist, without concurrently enhancing herbivore fitness. PMID- 11472772 TI - "Antioxidant defenses in caterpillars: role of the ascorbate-recycling system in the midgut lumen" by Raymond V. Barbehenn, Stacie L. Bumgarner, Erica F. Roosen, Michael M. Martin Journal of Insect Physiology 47 (4-5) pp. 349-357 (2001). PMID- 11472771 TI - History dependence of circadian pacemaker period in the cockroach. AB - The freerunning period of circadian clocks in constant environmental conditions can be history-dependent, and one effect of entrainment of circadian clocks by light cycles is to cause long-lasting changes in the freerunning period that are termed after-effects. We have studied after-effects of entrainment to 22-h (LD 8:14) and 26-h (LD 8:18) light cycles in the cockroach Leucophaea maderae. We find that in cockroaches, the freerunning period of the locomotor activity rhythm, measured in constant darkness (DD), is 0.7h less after entrainment to T22 than after entrainment to T26. Induction of after-effects requires several days (>1 week) entrainment, and after induction, after-effects will persist in DD for over 40 days. Further after-effects are unaltered by phase-resetting of up to 12h caused by exposure to low-temperature pulses (7 degrees C) of 24 or 48h duration. After-effects also persist through re-entrainment for 2 weeks to 24-h light cycles. These results indicate that after-effects arise from stable changes in the circadian system that are likely to be independent of phase relationships among oscillators within the circadian system. We also show that entrainment to temperature cycles does not generate after-effects indicating that light may be unique in its ability to generate lasting changes in pacemaker period. PMID- 11472773 TI - The role of membrane patch size and flow in regulating a proteolytic feedback threshold on a membrane: possible application in blood coagulation. AB - Positive feedback controls in proteolytic systems are characterized by thresholds which are regulated by the concentration of the initial stimulus and the kinetic parameters for enzyme generation and inhibition. Significant complexity is added when a positive feedback is localized on a membrane in contact with a flowing medium, such as seen in the early steps of blood coagulation. A partial differential equation model of an archetypal feedback loop is examined in which a proteolytic enzyme catalyzes its own formation from a zymogen on a membrane in contact with a flowing medium. As predicted from prior solution-phase and membrane-phase analyses, the threshold conditions for activation of the system are regulated by the kinetics of enzyme generation and inhibition and by the density of reactant-binding sites on the membrane; but the present analysis also establishes how the feedback threshold is controlled by the flow rate of the adjacent medium and the physical size of the membrane patch on which the feedback loop is localized. For given systems of particular kinetic properties, lower flow rates or larger active patches of membrane can result in the activation threshold being exceeded, whereas higher flow rates or smaller membrane patches can prevent initiation. In addition to numerical simulation, a simplified non-flowing model is analyzed to formulate an approximate mathematical statement of the dependence of the minimum activatable patch size on the kinetic and other parameters. PMID- 11472774 TI - Recovery of normal hemopoiesis in disseminated cancer therapy--a model. AB - The strategy of normal cell regeneration with recombinant hematopoietic growth factors during cancer chemotherapy is investigated by superimposing a treatment protocol on a simple model that describes an expanding malignant cell population that is coexisting with and inhibiting the population of normal cells. The model predictions suggest that the strategy of normal cell stimulation, possibly with growth factors, and possibly carried out within an intensive treatment framework may be a worthwhile chemotherapeutic option. Under this protocol, the model also predicts a minimum time interval for active treatment, a time to discontinue treatment, and a rest period during treatment in order to guarantee patient safety and recovery. Consequently, by relating and comparing model predictions to patient data, model simulations forecast that treatment could be shortened by 1-2 weeks if organized over or in the neighborhood of a predicted optimal time interval. Following this, it is conjectured that such an approach engendered by the model could produce outcomes that may have an edge over outcomes arising from therapeutic strategies that are executed over time frames that are relatively longer or significantly shorter than the predicted optimal time. PMID- 11472775 TI - On the firing maps of a general class of forced integrate and fire neurons. AB - Integrate and fire processes are fundamental mechanisms causing excitable and oscillatory behavior. Van der Pol [Philos. Mag. (7) 2 (11) (1926) 978] studied oscillations caused by these processes, which he called 'relaxation oscillations' and pointed out their relevance, not only to engineering, but also to the understanding of biological phenomena [Acta Med. Scand. Suppl. CVIII (108) (1940) 76], like cardiac rhythms and arrhythmias. The complex behavior of externally stimulated integrate and fire oscillators has motivated the study of simplified models whose dynamics are determined by iterations of 'firing circle maps' that can be studied in terms of Poincare's rotation theory [Chaos 1 (1991) 20; Chaos 1 (1991) 13; SIAM J. Appl. Math. 41 (3) (1981) 503]. In order to apply this theory to understand the responses and bifurcation patterns of forced systems, it is fundamental to determine the regions in parameter space where the different regularity properties (e.g., continuity and injectivity) of the firing maps are satisfied. Methods for carrying out this regularity analysis for linear systems, have been devised and the response of integrate and fire neurons (with linear accumulation) to a cyclic input has been analyzed [SIAM J. Appl. Math. 41 (3) (1981) 503]. In this paper we are concerned with the most general class of forced integrate and fire systems, modelled by one first-order differential equation. Using qualitative analysis we prove theorems on which we base a new method of regularity analysis of the firing map, that, contrasting with methods previously reported in the literature, does not requires analytic knowledge of the solutions of the differential equation and therefore it is also applicable to non-linear integrate and fire systems. To illustrate this new methodology, we apply it to determine the regularity regions of a non-linear example whose firing maps undergo bifurcations that were unknown for the previously studied linear systems. PMID- 11472776 TI - EcoGenomics--a consilience for comparative immunology? PMID- 11472777 TI - A screen of mammalian antibodies on snapper (Pagrus auratus, Sparidae) peripheral blood leukocytes reveals cross reactivity of an anti-human CD3 antibody with a population of mIg(-) cells. AB - Detailed immunological studies of the teleosts have been hampered by a lack of antibodies against cell-specific markers. Furthermore, where antibodies have been raised, in many instances they have been found to be species-specific. In comparison, many monoclonal and polyclonal antibodies exist with specificities for mammalian proteins and glycoproteins that effectively differentiate leukocyte sub-populations. In this study, we have tested a panel of 54 commercial antibodies against human and murine cell surface receptors for their ability to bind leukocytes isolated from the peripheral blood of snapper (Pagrus auratus). From this panel, one antibody, A452, which is specific for the intracytoplasmic tail of the epsilon (epsilon) chain of the T cell receptor-associated CD3 complex (CD3epsilon) bound to a subpopulation of peripheral blood leukocytes. Mutually exclusive counterstaining was observed when this antibody was used in conjunction with a monoclonal anti-snapper immunoglobulin antibody. This suggests that A452 may be binding to putative snapper T cells. PMID- 11472778 TI - Hematopoietic tissue allografts in Biomphalaria glabrata (Mollusca: Pulmonata) induce humoral immunity to Schistosoma mansoni. AB - We examined a potential mechanism for adoptively transferred resistance against Schistosoma mansoni in schistosome-susceptible snails receiving allografts of the hematopoietic amebocyte-producing organ (APO) from a schistosome-resistant strain of Biomphalaria glabrata. Susceptible NIH albino snails first were implanted with the APO from resistant Salvador strain donors. At 14 days post-implantation, cell free plasma was isolated from APO recipients and injected into a second group of NIH albino snails. Controls were injected with plasma from NIH albino snails that previously had received implants of Salvador mantle, an immunologically inert tissue. Finally, plasma recipients, along with untampered (non-injected) controls, were exposed to miracidia of S. mansoni and subsequently monitored for infection. A significantly lower prevalence of infection occurred in the group injected with plasma from APO recipients (53%) in comparison with that in untampered controls (100%) or in snails receiving plasma from mantle recipients (97%). These results suggest that adoptively transferred resistance in B. glabrata results from secretion of soluble resistance factor(s) by the implant, rather than, or in addition to, formation of a hemocyte chimera having cytotoxic donor hemocytes. PMID- 11472779 TI - Immune gene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific White Shrimp, Litopenaeus vannamei, and the Atlantic White Shrimp, L. setiferus. AB - A pilot program was undertaken in immune gene discovery in two sister species of litopenaeid shrimp, the Pacific white shrimp, Litopenaeus vannamei and the Atlantic white shrimp, L. setiferus. RNA from the hemocytes and hepatopancreas of single individuals from each species was recovered, 4 cDNA libraries (one from each tissue/species) were made by a PCR-based method and a total of approximately 2045 randomly selected clones were sequenced. A total of 268 expressed sequence tags (ESTs) were found that corresponded to 44 immune function genes. The most common immune-function ESTs (172) were antimicrobial peptides, which were restricted to the hemocyte libraries. Lectins were the largest group of immune function ESTs found in the hepatopancreas. Analysis of these libraries indicates that EST approaches are effective for immune gene discovery in shrimp and that the diversity of these PCR-generated libraries would support full-scale EST collection. PMID- 11472780 TI - Genomic organization and differential expression of channel catfish MHC class I genes. AB - Two clones, designated Icpu-UA/3 and Icpu-UA/26, were isolated from a genomic library prepared from a single homozygous gynogenetic channel catfish. Sequence analysis showed that each clone encoded a gene product containing features conserved among MHC class I molecules. The genomic organization of both clones indicated that each domain, with the exception of the cytoplasmic, was encoded by a separate exon. Moreover, like mammals, catfish cytoplasmic regions were encoded by three exons rather than two as previously described for other teleost MHC class I genes. Analysis of nucleotide sequences upstream of catfish class I genes revealed the presence of several regulatory motifs similar to those seen in mammalian class I genes. These included a TATA box, Enhancer B, Site alpha, ISRE, and GAS elements. To determine the functional significance of these elements, EMSAs and tissue expression assays were performed. EMSAs demonstrated that an Enhancer B element within Icpu-UA/26, and an imperfect Enhancer B element and/or a GC-rich region within Icpu-UA/3 were responsible for formation of specific DNA/protein complexes. Expression studies detected Icpu-UA/26 transcripts in all tissues tested, whereas Icpu-UA/3 encoded messages were seen in a limited number of tissues. These results define the intron/exon organization of catfish MHC class I genes, suggest that Icpu-UA/3 encodes a nonclassical gene, and provide the first functional evidence that upstream sequences, similar to those seen in mammalian class I genes, play important roles in regulating teleost MHC gene expression. PMID- 11472781 TI - Organization of the NKEF gene and its expression in the common carp (Cyprinus carpio). AB - Natural killer enhancing factor (NKEF) is a member of the newly defined peroxiredoxin (Prx) family. Its functions are to enhance the cytotoxic capacity of natural killer cells and to prevent DNA and protein from being damaged by oxidative stress in the presence of thiol compounds. However, little is known about the structure and function of NKEF in lower vertebrates. We have recently cloned a cDNA encoding NKEF from the common carp (Cyprinus carpio) by use of suppression subtractive hybridization (SSH). In the present study, we used PCR to obtain a genomic DNA which covers the entire coding region of carp NKEF. In the 3363bp-long genomic sequence, six exons and five introns were identified. The carp NKEF gene has splice donor/acceptor site sequences at the boundaries of exons and introns, and contains two Val-Cys-Pro (VCP) motifs. The exon/intron organization of the carp NKEF gene shows complete conservation with other members of the Prx family. Genomic Southern blotting analyses suggest that carp has multiple copies of the NKEF gene. RT-PCR analyses reveal that carp NKEF has very different expression levels not only in tissues but also from individuals. PMID- 11472782 TI - Location of superoxide production sites in turbot neutrophils and gilthead seabream acidophilic granulocytes during phagocytosis of glucan particles. AB - Here we show, by spectrophotometry and enzyme cytochemical methods, that turbot neutrophils and gilthead seabream acidophils responded in a similar way when incubated with PMA or with particulate glucans. Cells stimulated with PMA released high amounts of superoxide both intra- and extracellularly. However, O2- was mainly released intracellularly when cells were incubated with particulate glucans. Small glucan particles were quickly phagocytosed and O2- was initially produced in intracellular vesicles and tubular structures that later fused with the phagosome or with the cell membrane. Large glucan filaments that were not phagocytosed also induced cell stimulation and O2- was also produced in intracellular vesicles which then appeared to fuse with the cell membrane. We conclude that, in stimulated turbot neutrophils and gilthead seabream acidophils, superoxide production is carried out initially in intracellular compartments that are very similar to those described in mammalian neutrophils. PMID- 11472783 TI - A fish cell surface receptor defined by a mAb mediates leukocyte aggregation and deactivation. AB - Cell adhesion molecules play a key role in the inflammatory response. Selectins, integrins and immunoglobulin gene superfamily adhesion receptors mediate the different steps of leukocyte migration from the blood-stream towards inflammatory foci. In addition to their adhesive function, these receptors modulate major cellular processes such as cell activation, growth, differentiation and death. To characterise the fish molecules involved in cell adhesion, a panel of mAbs was raised by immunising mice with macrophages from the marine fish gilthead seabream (Sparus aurata L.). One of these mAbs, which we named anti-Aggregatin, was found to induce a rapid heterotypic aggregation of seabream leukocytes. Anti-Aggregatin defined a 140-kDa cell surface receptor which was highly expressed by macrophages and was up-regulated after co-stimulation with LPS and MAF. Functionally, the cell adhesion which occurred upon exposure to anti-Aggregatin required Ca(2+), an intact cytoskeleton and an active cell metabolism. More importantly, Aggregatin engagement resulted in strong inhibition of the phagocyte respiratory burst, although the cells showed neither loss of viability nor DNA fragmentation. The results are discussed in relation to the potential role of cell adhesion molecules in fish immune responses. PMID- 11472784 TI - Divergence of the inflammatory response in two types of chickens. AB - We compared inflammatory responses to lipopolysaccharide (LPS) injection in laying type (Brown Nick) to broiler type (Avian x Avian) chicks. Rectal temperature was measured at 0, 1, 2, 4, 6, 12, and 24h after LPS injection (0, 0.1, 0.3, 0.6, 1, 2.5, or 5mg/kg bw). In layers, rectal temperature increased from 41.31+/-0.19 degrees C to a maximum 42.27+/-0.41 degrees C at 4h after 1mg/kg LPS. Relative to layers, the febrile response in broilers was considerably lower, delayed in onset, and required higher levels of LPS (5mg/kg). Proliferation of spleen cells from un-injected chicks in response to LPS, PHA, and Con A was evaluated in vitro. IFNgamma, TGFbeta(2), MGF and IL-1beta relative to beta-actin mRNA expression were analyzed in spleen cells stimulated with LPS. Splenocytes from layers had a higher proliferative response to LPS (P=0.045), but lower proliferative response to PHA (P=0.004) and Con A (P=0.004) than broilers. Expression of mRNA for MGF, IL-1beta and IFNgamma was lower in broilers than in layers (P<0.001). Reduced production of the pro-inflammatory cytokines in broilers could have resulted from the observed increased production of the immunosuppressive cytokine TGFbeta(2.) These differences in cytokine expression may explain the blunted febrile response in broilers compared to layers. Because the acute phase response of inflammation causes decreased food intake, the blunted inflammatory response of broilers may permit faster growth. PMID- 11472785 TI - Signal transduction pathways activated by engaging immunoglobulin Fc receptors on chicken heterophils. AB - In the present studies, we initiated experiments to identify the signal transduction factors involved in activating phagocytosis, oxidative burst, and degranulation following the binding of IgG-opsonized SE to Fc receptors on the surface of avian heterophils. Peripheral blood heterophils were isolated and exposed to known inhibitors of signal transduction pathways for either 20min (chelerythine, genistein, or verapamil) or 120min (pertussis toxin) at 39 degrees C. The cells were then stimulated for 30min at 39 degrees C with SE opsonized with IgG purified from SE-immune chickens. Phagocytosis, luminol-dependent chemiluminescence (LDCL), and beta-D-glucuronidase release were then evaluated in vitro. The G-protein inhibitor, pertussis toxin, the protein kinase C inhibitor, chelerythine, and the Ca(++) channel blocker, verapamil, markedly reduced phagocytosis in a dose responsive manner. Genistein, a tyrosine kinase inhibitor, had no effect on the phagocytosis of the opsonized SE. Both pertussis toxin (66 98%) and verapamil (47-76%) had marked inhibitory effect on LDCL. Chelerythine (13-25%) and genistein (5-25%) had far less biologically significant effects on LDCL. Neither chelerythine nor genistein had a significant effect on degranulation. Verapamil (2-28%) and pertussis toxin (25-29%) had a moderate inhibitory effect on degranulation stimulated by IgG-opsonized SE. As was found with complement receptor mediated activation of heterophils, the binding of Fc receptors by the IgG-SE complex activated distinct signaling pathways that regulate the functional activities of avian heterophils. Pertussis toxin sensitive, Ca(++)-dependent, G-proteins and protein kinase C-dependent protein phosphorylation play a major role in the phagocytosis of IgG-opsonized SE. Pertussis toxin-sensitive, Ca(++)-dependent, G-proteins appear to regulate LDCL following Fc receptor binding. The signal transduction inhibitors used in these studies did not affect Fc receptor mediated degranulation by avian heterophils. PMID- 11472786 TI - Fluvoxamine in the treatment of panic disorder: a multi-center, double-blind, placebo-controlled study in outpatients. AB - Serotonergic mechanisms have been implicated in panic disorder, and several preliminary studies suggest that fluvoxamine, a selective serotonin reuptake inhibitor (SSRI), is helpful in its treatment. This 8-week double-blind parallel group study compared fluvoxamine with a placebo in 188 patients with DSM-III-R defined panic disorder with or without agoraphobia. Efficacy assessments included a Daily Panic Attack Inventory, the Sheehan Disability Scale, the Clinical Anxiety Scale and the Clinical Global Impression Scale. When compared with the placebo, fluvoxamine produced highly significant improvements in most measures of the frequency and severity of panic disorder and in the more global aspects of disability and distress. Significant improvement was evident as early as week 1 for some panic variables. Fluvoxamine is a potent anti-panic agent with a relatively rapid onset of action. PMID- 11472787 TI - Self-destructiveness and serotonin function in bulimia nervosa. AB - Studies have linked bulimia nervosa (BN) to alterations in brain serotonin (5 hydroxytryptamine: 5-HT) activity and to heightened propensity for parasuicidality and self-injuriousness. The coincidence of self-destructiveness and 5-HT abnormality in BN is of interest, given documentation (in various populations) of an inverse association between 5-HT activity and potential for self-harm. The present study examined the connection between 5-HT status and self destructiveness in BN. Structured interviews and self-report questionnaires were used to assess 40 bulimic and 21 normal-eater women for: (a) history of parasuicidal or self-injurious acts; and (b) mood and impulse-regulation problems. We then applied tests, presumed to reflect 5-HT function, of serial prolactin (PRL) and cortisol (CORT) responses after oral administration of the partial 5-HT agonist, meta-chlorophenylpiperazine (m-CPP). Relative to non bulimic women, bulimic women (on average) showed blunting of PRL and CORT following m-CPP. The blunting of neuroendocrine responses was, however, most remarkable in bulimic women with a history of self-destructiveness. These findings suggest that some serotonergic anomalies reported in BN sufferers (i.e. reduced neuroendocrine response after m-CPP) may be most characteristic of individuals in the population showing clear-cut self-destructive potential. PMID- 11472788 TI - Flunitrazepam abuse and personality characteristics in male forensic psychiatric patients. AB - Sixty male non-psychotic forensic psychiatric patients (aged 16-35 years) were studied after they completed their ordinary forensic psychiatric assessment (FPA). The prevalence of flunitrazepam (FZ) abuse was investigated by using both structured and in-depth interviews with the objective of studying the relationship between the abuse and personality traits. The patient's characteristics, DSM-IV disorders, and actual sentences were obtained by studying their files. In order to obtain measures on their personality traits, self-report inventories were administered to the patients. Eighteen out of 60 patients were FZ abusers, but only 4 of them received a diagnosis related to the FZ abuse during the ordinary FPA. In almost all cases, however, indications of the FZ abuse were found in the files. No differences in personality traits were found between the groups. The frequency of previous admissions to an FPA and actual sentences of robbery, weapons offenses, narcotic-related offenses, and other crimes (such as theft) among the FZ abusers deviated significantly from forensic non-FZ abusers. Therefore, the FZ abuse per se might be more responsible for their tendency to commit crimes characterized by danger and thrill-seeking (such as robbery, weapons offences, and theft) than personality. The most important conclusion is that assessment of FZ abuse is needed in forensic psychiatry. PMID- 11472789 TI - Pedophilia is accompanied by increased plasma concentrations of catecholamines, in particular epinephrine. AB - Plasma epinephrine and norepinephrine concentrations were measured in pedophiles and normal men both in placebo conditions and after administration of meta chlorophenylpiperazine (mCPP), a post-synaptic 5-HT2 receptor agonist. The plasma concentrations of catecholamines, in particular epinephrine, were significantly increased in pedophiles. It is concluded that pedophiles may have an increased activity of the sympathoadrenal system. PMID- 11472790 TI - Sleep and treatment response in depression: new findings using power spectral analysis. AB - This study examined quantitative measures of sleep electroencephalogram (EEG) and phasic rapid eye movements (REM) as correlates of remission and recovery in depressed patients. To address correlates of remission, pre-treatment EEG sleep studies were examined in 130 women outpatients with major depressive disorder treated with interpersonal psychotherapy (IPT). To address correlates of recovery, baseline and post-treatment EEG sleep studies were examined in 23 women who recovered with IPT alone and 23 women who recovered with IPT+fluoxetine. Outcomes included EEG power spectra during non-rapid eye movement (NREM) sleep and REM sleep and quantitative REMs. IPT non-remitters had increased phasic REM compared with remitters, but no significant differences in EEG power spectra. IPT+fluoxetine recoverers, but not IPT recoverers, showed increases in phasic REM and REM percentage from baseline to recovery. In NREM sleep, the IPT+fluoxetine group showed a decrease in alpha power from baseline to recovery, while the IPT group showed a slight increase. The number of REMs was a more robust correlate of remission and recovery than modeled quantitative EEG spectra during NREM or REM sleep. Quantitative REMs may provide a more direct measure of brainstem function and dysfunction during REM sleep than quantitative sleep EEG measures. PMID- 11472791 TI - Health-related quality of well-being in chronically hospitalized patients with schizophrenia: comparison with matched outpatients. AB - Quantifying the functional consequences of illness in terms of quality of life can enhance our understanding of both mental and physical disorders. However, little is known about the quality of life among older inpatients vs. outpatients with schizophrenia. We present the results of health-related quality of life assessments in 54 middle-aged and elderly long-term inpatients with schizophrenia and a demographically matched outpatient sample. Assessments were performed using the Quality of Well-Being (QWB) scale, along with standard measures of psychopathology and global cognitive impairment. Compared with outpatients, the inpatients had a significantly lower health-related quality of life, as measured by the QWB. In the inpatient and outpatient groups, higher levels of positive symptoms were associated with lower health-related quality of life. Health related quality of life remained fairly stable among the inpatients who remained hospitalized over 6 months. In both inpatients and outpatients, baseline cognitive status and psychopathology predicted QWB scores at the 6-month follow up. These findings further support the use of the QWB in severely mentally ill populations; implications for improving health-related quality of life among older patients with schizophrenia are discussed. PMID- 11472792 TI - Tryptophan hydroxylase gene and major psychoses. AB - Disturbances of the serotoninergic neurotransmitter system have been implicated in the pathogenesis of mood disorders. The tryptophan hydroxylase (TPH) gene, which codes for the rate-limiting enzyme of serotonin biosynthesis, has been recently reported to be associated with bipolar disorder. In this study, we investigated TPH A218C gene variants in a sample of subjects affected by major psychoses. One thousand four hundred and twenty-four inpatients affected by bipolar (n=627), major depressive (n=511), schizophrenic (n=210), delusional (n=48) disorder and psychotic disorder not otherwise specified (n=27) (DSM-IV) were included; all patients and 380 controls were typed for the TPH variants using PCR techniques. A sub-sample of 963 patients was assessed using the Operational Criteria for Psychotic Illness (OPCRIT). TPH variants were not associated with major psychoses, but a trend was observed toward an excess of TPH*A/A in bipolar disorder. The analysis of symptomatology factors did not show any significant difference either; however, a trend was observed for males with the TPH*A genotype to have lower depressive symptoms compared with TPH*C subjects. Possible stratification factors such as current age and age of onset did not affect the observed results. TPH A218C variants are not, therefore, a major liability factor for the symptoms of major psychoses to have in the present sample. TPH*A containing variants may be a protective factor for depressive symptoms among male subjects with mood disorders or for a subtype of mood disorders characterized by a mainly manic form of symptomatology. PMID- 11472793 TI - Frequency and severity of religious delusions in Christian patients with psychosis. AB - The influence of religious activity on the severity of religious delusions is unclear. This study examined whether Catholic and Protestant patients experienced more religious delusions than non-religiously affiliated patients. We also explored whether the severity of religious delusions, according to the Religious Delusions item on the Scale for the Assessment of Positive Symptoms (SAPS), was associated with the amount of religious activity. The Protestants experienced more religious delusions than Catholics and those without religious affiliation. Although when the groups were combined, patients who were more religiously active experienced more severe religious delusions (n=133), there was no difference in the severity of religious delusions across the non-religious, Catholic and Protestant groups. Religious affiliation may influence the frequency of religious delusions, particularly in Protestant individuals, but religious affiliation appears to be independent of religious delusion severity. PMID- 11472794 TI - Biochemical markers of anxiety and depression. AB - The results of research into a biochemical marker for depression are presented. The research was carried out on a normal population in a primary care setting. Cholesterol levels were identified as the blood marker for anxiety and depression. The Hamilton Rating Scale for Depression was chosen to identify depression. Those with low cholesterol scored significantly higher on the Hamilton Rating Scale for Depression. PMID- 11472795 TI - Monitoring for adsorption of human serum albumin and bovine serum albumin onto bare and polystyrene-modified silver electrodes by quartz crystal impedance analysis. AB - The adsorption of human serum albumin (HSA) and bovine serum albumin (BSA) from PBS (pH 7.4) onto bare and polystyrene (PS)-modified silver electrodes was in situ monitored using quartz crystal impedance analysis. The adsorption characteristics of HSA and BSA were discussed by analyzing piezoelectric parameter simultaneous responses. Experimental results indicated that for both HSA and BSA, the amount adsorbed on bare silver was more than that on PS-modified surface. The BSA amount adsorbed on the two surfaces was more than that of adsorbed HSA. A kinetic model was developed to describe the adsorption process and fitted to the experimental data of frequency shift. It was shown that HSA adsorption could be described by a kinetic equation involving two consecutive reactions. At lower concentration, BSA adsorption only involved the first reaction. At higher concentration, BSA adsorption on PS-modified surface involved two consecutive reactions. All fitted results were well in agreement with the corresponding experimental results. The regression values of reaction rate constants for the HSA and BSA adsorption were obtained. These data exhibited difference in adsorption kinetics under different conditions. PMID- 11472796 TI - A novel esterase from Burkholderia gladioli which shows high deacetylation activity on cephalosporins is related to beta-lactamases and DD-peptidases. AB - The gene (estB) encoding for a novel esterase (EstB) from Burkholderia gladioli (formerly Pseudomonas marginata) NCPPB 1891 was cloned in Escherichia coli. Sequence analysis showed an open reading frame encoding a polypeptide of 392 amino acid residues, with a molecular mass of about 42 kDa. Comparison of the amino acid sequence with those of other homologous enzymes indicated homologies to beta-lactamases, penicillin binding proteins and DD-peptidases. The serine residue (Ser(75)) which is located within a present class A beta-lactamase motif ([F,Y]-X-[L,I,V,M,F,Y]-X-S-[T,V]-X-K-X-X-X-X-[A,G,L]-X-X-[L,C]) was identified by site-directed mutagenesis to represent the active nucleophile. A second serine residue (Ser(149)) which is located within a G-x-S-x-G motif which is typically found in esterases and lipases was demonstrated not to play a significant role in enzyme function. The estB gene was overexpressed in E. coli using a tac promoter based expression system. Investigation of EstB protein with respect to the ability to hydrolyse beta-lactam substrates clearly demonstrated that this protein has no beta-lactamase activity. The recombinant enzyme is active on triglycerides and on nitrophenyl esters with acyl chain lengths up to C6. The preference for short chain length substrates indicated that EstB is a typical carboxylesterase. As a special feature EstB esterase was found to have high deacetylation activity on cephalosporin derivatives. PMID- 11472797 TI - Optimization of citric acid production from Candida lipolytica Y-1095 using n paraffin. AB - Currently, the majority of worldwide microbial production of citric acid utilizes Aspergillus niger in a carbohydrate based submerged fermentation. Due to their high carbon content, hydrocarbons also have the potential of producing high concentrations of citric acid. Initial lab experiments conducted using 1875 ml batch fermentations with n-paraffin found that Candida lipolytica NRRL-Y-1095 assimilated the feedstock and had a citric acid productivity of 47 mg l(-1) h( 1). To determine the optimum level of initial biomass concentration, n-paraffin concentration, iron concentration and temperature for the production of citric acid, a central composite design was developed using 200 ml batch fermentations. The design involved conducting 31 batch fermentations under various combinations of high and low values of these four parameters. From this investigation empirical models were developed describing the interactions between the experimental parameters and citric acid production. It was found that the maximum concentration of citric acid produced was 9.8 g l(-1) and the optimum levels of each parameter for citric acid production were, 10--12% volume for initial biomass concentration, 10--15% volume for n-paraffin concentration, 10 mg l(-1) for ferric nitrate concentration, and 26--30 degrees C for temperature. PMID- 11472798 TI - Use of dynamic step response for control of fed-batch conversion of lignocellulosic hydrolyzates to ethanol. AB - Optimization of fed-batch conversion of lignocellulosic hydrolyzates by the yeast Saccharomyces cerevisiae was studied. The feed rate was controlled using a step response strategy, in which the carbon dioxide evolution rate was used as input variable. The performance of the control strategy was examined using both an untreated and a detoxified dilute acid hydrolyzate, and the performance was compared to that obtained with a synthetic medium. In batch cultivation of the untreated hydrolyzate, only 23% of the hexose sugars were assimilated. However, by using the feed-back controlled fed-batch technique, it was possible to obtain complete conversion of the hexose sugars. Furthermore, the maximal specific ethanol productivity (q(E,max)) increased more than 10-fold, from 0.06 to 0.70 g g(-1) h(-1). In addition, the viability of the yeast cells decreased by more than 99% in batch cultivation, whereas a viability of more than 40% could be maintained during fed-batch cultivation. In contrast to untreated hydrolyzate, it was possible to convert the sugars in the detoxified hydrolyzate also in batch cultivation. However, a 50% higher specific ethanol productivity was obtained using fed-batch cultivation. During batch cultivation of both untreated and detoxified hydrolyzate a gradual decrease in specific ethanol productivity was observed. This decrease could largely be avoided in fed-batch cultivations. PMID- 11472799 TI - Polysaccharide synthesis as a carbon dissipation mechanism in metabolically uncoupled Xanthomonas campestris cells. AB - The utilization of xanthan metabolism as an excess carbon dissipation path in Xanthomonas campestris cells under sub-lethal acid stress was studied. To highlight growth limitation during metabolic uncoupling due to acid toxicity a antibiotic was added. The simultaneous addition of enoxacin and acetic acid showed that the xanthan production per unit of biomass raises with increasing concentrations of enoxacin, which seems to indicate that when the cell is prevented from growing it finds a path to convey the extra carbon. In parallel, although the effect of acetic acid is not very significant, its presence appears to increase xanthan. This tendency seems to be accentuated with increasing concentrations of enoxacin. In fact, in presence of 0.15 mM of acetic acid, 2.88 and 5.76 microM of antibiotic produces xanthan/biomass yields of 8.13 and 9.82 g g(-1) which drop to below half those values (3.55 g g(-1)) when enoxacin is removed. When enoxacin was kept constant, xanthan/biomass yields showed small increments with the increase of acetic acid. Thus, with 1.44, 2.88 and 4.32 microM enoxacin concentrations, the addition of organic acid produces a 6--8% stimulation of xanthan. PMID- 11472800 TI - Two-stage cultures for the production of astaxanthin from Haematococcus pluvialis. AB - A two-stage culture system was established for the production of astaxanthin from Haematococcus pluvialis. In a first stage green vegetative cells were produced in semicontinuous cultures maintained with daily renewal rates between 10 and 40%. The steady-state cell density decreased with increasing renewal rates. Highest cell productivity, 64 x 10(6) cells l(-1) day(-1) was obtained with a daily renewal rate of 20%. In a second stage the harvested cultures were submitted to high light (240 micromol photon m(-2) s(-1)) under batch conditions for 15 days in order to stimulate the transition to the aplanospore stage and the accumulation of astaxanthin. No decrease in cell density was recorded during the induction period in any of the cultures. Cultures obtained at high renewal rates continued growing during the induction period and no astaxanthin was accumulated until all nitrogen in the media had been consumed. The final concentration of astaxanthin was inversely correlated to the growth rate at which first-stage cultures were maintained. Optimal renewal rate for maximal astaxanthin production depended on the duration of the induction period. After a 12-day induction period the highest astaxanthin production, 5.8 mg l(-1) of semi-continuous culture day 1, was obtained with cultures maintained at a renewal rate of 20%. When the induction period was increased to 15 days maximal astaxanthin productivity, 9.6 mg l(-1) of semi-continuous culture day -1, was obtained from cultures maintained at a renewal rate of 40% despite the much lower astaxanthin concentration achieved in these cultures. Results demonstrate the feasibility of semi continuous cultivation of H. pluvialis for the two-stage production of astaxanthin. PMID- 11472801 TI - A method to maintain introduced DNA sequences stably and safely on the bacterial chromosome: application of prophage integration and subsequent designed excision. AB - By application of prophage integration and subsequent intended excision, a method to maintain an introduced DNA sequence stably onto a bacterial chromosome has been proposed. Recently-constructed integration plasmids using Campbell-type prophage integration system in Lactobacillus casei strain Shirota and its temperate phage phi FSW was modified for this purpose and a chloramphenicol (Cm) resistance gene was used as a model passenger DNA. On the integration plasmid having an erythromycin (Em)-resistance gene as a selection marker, N- and C terminally-truncated Cm-resistance genes were inserted into both sides of the attP of phi FSW, within which the site-specific recombination took place with the attB of phi FSW on the recipient chromosome through the phi FSW integrase. Primary integrants of the modified plasmid (integration-excision vector) exhibiting Em-resistant and Cm-sensitive phenotype generated Em-sensitive and Cm resistant derivatives under the nonselective conditions. Sequence analyses showed that one copy of the complete Cm-resistance gene resided at the attachment site on the host chromosome and the other vector-derived sequences were excised probably by endogenous homologous recombination in the host cells to derive final integrants. The Cm-resistant phenotype of the final integrants was stable for more than 50 generations under non-selective conditions. Frequency of the homologous recombination suggests that negative selection is also adoptable. Thus, this method using the integration-excision vector gives a stable and safe derivatives of the strain and is likely to be applicable to various bacteria, since Campbell-type prophage integration system and homologous recombination are prevalent among bacteria. PMID- 11472802 TI - Agar polysaccharides from Gracilaria species (Rhodophyta, Gracilariaceae). AB - Yield, physical and chemical properties of agar from three agarophytes species (Gracilaria gracilis, G. dura and G. bursa-pastoris) were determined. The agar yield from the three species varied significantly (P<0.01). The highest yields of agar (34.8%) and the lowest (30%) were obtained from G. bursa-pastoris and G. gracilis, respectively. Highest gel strength (630+/-15 g cm(-2)) was obtained from agar extracted from G. gracilis and lowest from G. bursa-pastoris (26+/-3.6 g cm(-2)). The values of 3,6-anhydrogalactose were similar for G. gracilis and G. dura and there were no significant differences among the species. The sulfate contents varied significantly (P<0.01) and the higher value was obtained from G. bursa-pastoris. Among the three species, G. gracilis showed superior agar quality than the other two species, hence it can be considered a good potential source for industrial use. PMID- 11472803 TI - Influence of patient factors on age of restorations at failure and reasons for their placement and replacement. AB - OBJECTIVE: This study examined the reasons given by a selected group of dental practitioners for placement and replacement of restorations and correlated the data provided with patient factors, such as patients' age and gender, caries risk, occlusal function and oral hygiene, with restoration longevity. METHOD: A group of general dental practitioners (GDPs) were recruited to take part in the study. Each participant was asked to record the reason for placement or replacement of restorations from a list of potential reasons. The age and Class of the restoration being replaced was also recorded, as also was the material being used and the material being replaced. RESULTS: Details of reason for placement/replacement was received on 3196 restorations from 32 GDPs. Of the restorations placed, 54% were amalgam, 32% composite, 8% compomer and 7% glass ionomer. The reasons for placement/replacement of the restorations were principally primary caries (28%), secondary caries (29%), margin fracture (10%), tooth fracture (7%), and non-carious defects (6%). Overall, the mean age of restorations at failure was 7.1 years. Of the patients who received glass ionomer restorations, 29% were rated as having poor oral hygiene, compared with 18% of the patients who received amalgam restorations, 18% of the patients who received composite restorations and 23% of the patients who received compomer restorations. Of the patients who received glass ionomer restorations, 35% were rated as having high caries susceptibility, compared with 27% of those receiving amalgam restorations, 21% of those receiving composite restorations and 30% of those receiving compomer restorations. CONCLUSION: Primary caries was the principal reason for initial restorations. Secondary caries was the most prevalent reason for replacement of restorations. The results also indicate a selective application of different materials for different patients. PMID- 11472804 TI - Does social deprivation in 1, 2, 3 and 4-year-old Scottish infants influence the frequency isolation of caries-associated micro-organisms? AB - OBJECTIVES: The aim of this study was to determine if there are associations between the level of social deprivation/affluence and the frequency isolation of caries-associated micro-organisms (Streptococcus mutans, Streptococcus sobrinus, lactobacilli and yeasts) in a large cohort of infants examined annually from 1 to 4 years of age. METHODS: DEPCAT was used to measure the socio-economic status of all consented infants (n=1099--1392) born in Dundee during a 1 year period (total n=1974). Caries-associated micro-organisms were cultured from saliva when the infants were 1, 2, 3 and 4 years of age. Standardised dental examinations were also carried out annually. Log linear analysis, which controlled for caries, was used to look for associations between DEPCAT and the isolation frequency of caries-associated micro-organisms. RESULTS: When controlling for caries, there was an association between DEPCAT and the isolation frequency of yeasts when the infants were 1 and 2 but not when 3 and 4 years old, whereas lactobacilli were associated only when the infants were 3 and 4 years old. Correlations between S. mutans and social deprivation were usually dependent on the caries status of the infants. CONCLUSIONS: The relationship between social deprivation and the isolation frequencies of caries-associated micro-organisms is complex with lactobacilli developing an association when the infants were 3 and 4 years old in contrast to yeasts which were only associated when the infants were 1 and 2 years old. Streptococcus mutans was associated with social deprivation when the infants were 2 years old and older, but dependent on caries status in the 3 and 4 year olds. PMID- 11472805 TI - Dental erosion in deciduous teeth--an in vivo and in vitro study. AB - OBJECTIVE: The aims of this study were to report on severe dental erosion and its associated etiological factors in deciduous teeth of Saudi children (n=16, X macro=6.5 years, R=5--8) in vivo, and to confirm (or reject) the clinical diagnosis by scanning electron microscopy (SEM). Another aim was to study progression of erosion in vitro using extracted healthy deciduous and permanent teeth. METHOD: For the in vivo study, a questionnaire and clinical examination was completed, medical history obtained and exfoliated eroded teeth (n=8) examined by SEM. For the in vitro study, enamel specimens were prepared from Saudi (n=21) and Finnish (n=19) deciduous teeth as well as Finnish permanent teeth (n=20), immersed in 2% citric acid for 5--30 min, microhardness measurements performed, and the specimens studied by SEM. RESULTS: The in vivo investigation showed that high intake of acidic drinks and fruits, upper respiratory tract problems and frequent medications may constitute possible etiological and/or aggravating factors for severe dental erosion in Saudi children. Deciduous tooth enamel was softer than permanent tooth enamel. SEM showed no clear differences in the ultrastructure of the enamel specimens amongst the three groups. The clinical diagnosis of severe dental erosion in those cases studied was reliable as verified by SEM. CONCLUSIONS: The presence of dental erosion in children is likely to be associated with a number of general health and dietary factors but is also aggravated by the relatively more rapid progression of erosion in the deciduous teeth. PMID- 11472806 TI - Transdentinal stimulation of reactionary dentinogenesis in ferrets by dentine matrix components. AB - OBJECTIVES: Trans-dentinal stimulation of reactionary dentinogenesis may be mediated through cellular signalling by bio-active components released from the dentine matrix during injury. Understanding of these processes will be important to guide dentinal repair activity following restorative surgery. The purpose of this study was to investigate the effects of implanting isolated dentine matrix proteins within cavity preparations on dentinal repair activity and odontoblast survival using a controlled experimental animal model. METHODS: Forty-five ferret canine teeth each had a standardised non-exposed cylindrical Class V cavity cut into the buccal dentine. Ten cavities were restored with zinc oxide eugenol, as a control. Two different lyophilised preparations of dentine matrix components were implanted on the axial floor of the remaining 35 cavities prior to filling with zinc oxide eugenol. After post-operative periods of 2, 7, 14, 28, and 90 days, the teeth were extracted and examined histomorphometrically and the data analysed statistically by analysis of variance tests. RESULTS: The odontoblasts beneath the restored cavities responded to the presence of the two dentine matrix preparations by increasing the mean area of reactionary dentine secreted by 433 and 578%, and the numbers of odontoblasts remained stable. CONCLUSION: Dentine matrix components can stimulate reactionary dentinogenesis in non-exposed cavity preparations. It will now be important to identify how this may be harnessed as a part of routine restorative surgery to optimise treatment outcomes with a biological basis. PMID- 11472807 TI - Free-electron laser etching of dental enamel. AB - OBJECTIVE: The purpose of this study was to evaluate the Mark-III free-electron laser as a means of etching enamel surfaces, with potential application to resin bonding. METHODS: The FEL was tuned to wavelengths ranging from 3.0 to 9.2 microm. Specific wavelengths that are resonantly absorbed by phosphates, proteins, and water were used. First, bovine enamel was polished and exposed to static FEL exposures. Lased enamel was examined using scanning electron microscopy (SEM). Additional bovine enamel specimens were exposed to FEL at similar wavelengths, but with rastering to create treated rectangular areas on each specimen. Surface roughness was evaluated using profilometry and atomic force microscopy (AFM). Composite was bonded to the lased enamel, and shear bond strengths were determined using an Instron universal testing machine. As a control, the surface roughness of, and shear bond strengths to, acid-etched enamel were determined. RESULTS: Static FEL exposures caused changes in the enamel ranging from an etched appearance to pits, cracks, and frank cratering. The surface roughness of lased enamel was much greater than that of acid-etched enamel, and was qualitatively different as well. Shear bond strengths of resin to acid-etched enamel were significantly higher than bond strengths to lased enamel. CONCLUSIONS: Under the conditions used in this study, the FEL did not offer a practical and effective method of etching enamel for resin bonding. However, the ability of the FEL to deliver many specific wavelengths makes it an interesting tool for further research of laser effects on tooth structure. PMID- 11472808 TI - In vitro analysis of the effects of acid or laser etching on microleakage around composite resin restorations. AB - OBJECTIVES: The aim of the present study was to determine the effect of two conditioning methods, phosphoric acid and XeCl laser application on microleakage at composite resin restorations. METHODS: Class II MOD (mesio-occluso-distal) cavities were prepared on 90 extracted human wisdom teeth with one proximogingival margin on enamel and the other on dentin. Phosphoric acid (37%) and a XeCl excimer laser system were used for the etching procedure. The prepared teeth were randomly assigned to six groups: In group 1, acid-etching of the enamel margins was followed by application of enamel bonding, in group 2 additionally a dentin bonding system was used. In group 3, acid-etching of the enamel and dentin (total etch) was followed by the application of dentin bonding. In group 4, laser-etching of enamel margins was followed by the application of enamel bonding, in group 5 dentin bonding was used additionally. In group 6, laser-etching of enamel and dentin margins was followed by the application of dentin bonding. All restorations were placed incrementally and polymerized with a halogen light source. After restoration, all samples were thermocycled and stained in 0.5% basic fuchsin and sectioned longitudinally in a mesio-distal plane. Microleakage was scored by means of a stereomicroscope. The margins of the restorations at tooth-resin interface were examined with a scanning electron microscope. RESULTS: In all groups, marginal leakage was more extensive at the gingival margin that was in dentin than at the gingival margin in enamel. Microleakage scores and the gap at the tooth-resin interface were significantly lower in all acid-etched groups than in laser-etched groups. CONCLUSION: Acid etch technique is more effective than the investigated laser-etch technique at 308nm concerning the reduction of marginal leakage in composite restorations. PMID- 11472809 TI - An investigation of cuspal deformation and delayed recovery after occlusal loading. AB - OBJECTIVE: The purpose of this study was to investigate the extent of progressive cuspal displacement during prolonged occlusal loading (either continuous or cyclic) and delayed cuspal recovery following removal of the load. METHODS: Extracted maxillary premolars with extensive MOD cavity plus endodontic access were subjected to an occlusal load applied to both cusps, followed by a recovery period. Two types of loading were applied: (a) continuous load of 100, 200 and 300 N for 0.1--1000 s; (b) cyclic loading of 300 N at 0.83 Hz for 1--1000 cycles. Cuspal displacement during loading and subsequent recovery were measured with an extensometer attached to both cusps. RESULTS: Continuous loading led to progressive cuspal displacement that was both time- and load-dependent. After removal of the load, complete recovery from cuspal deformation was prolonged up to 20 min, also in a time- and load-dependent manner. Cyclic loading resulted in a cumulative increase in cusp displacement but to only a very small extent (approximately 1 microm after 1000 cycles). CONCLUSIONS: Progressive cuspal displacement and delayed recovery occur with prolonged continuous or cyclic occlusal loading. Continuous loading (e.g. as in clenching) is potentially more damaging than cyclic loading (as in chewing). PMID- 11472810 TI - In vitro remineralisation of eroded enamel lesions by saliva. AB - OBJECTIVES: It is speculated that saliva, with its mineral content, may possess a reparative effect on an early erosion which is characterised by softened surface and slight subsurface demineralisation in addition to a crater. This study aimed to determine the possible remineralisation of early enamel erosion by saliva. METHODS: Eroded lesions were produced in bovine incisors by 1-h immersion in orange juice. Control sections and three experimental slabs were produced from each tooth. The three slabs were assigned randomly to one of three remineralising agents: clarified natural saliva (NS), artificial saliva (AS) and remineralising solution (RS). All solutions had a pH of 7.2, a fluoride concentration of 0.022 ppm, and were changed daily. NS was collected daily from the same individual at the same time of day. The specimens were exposed to their respective remineralising agents for 28 days. Using microradiography and image analysis, the mineral loss (Delta z) and lesion depth (ld) were quantified in sections cut from the control and experimental slabs. RESULTS: A significant (p<0.001) amount of mineral was gained following exposure to each remineralising agent. Significantly less Delta z and ld were observed for the experimental groups compared with the control group (p<0.001; paired t-test). This effect was greatest with RS and least with AS. Inter-group comparison (Duncan multiple tests) showed no significant difference in Delta z among the experimental groups, however ld was significantly higher for AS (p<0.001) compared with RS and NS, and no difference was observed between RS and NS. CONCLUSION: Saliva as well as remineralising solutions can remineralise early enamel erosion. PMID- 11472811 TI - Reinforcement effect of short glass fibers with CaO- P(2)O(5) -SiO(2) -Al(2)O(3) glass on strength of glass-ionomer cement. AB - OBJECTIVES: A high strength glass-ionomer (not resin-modified) cement was developed using short fibers of CaO--P(2)O(5)--SiO(2)--Al(2)O(3) (CPSA) glass. The purpose of this study was to clarify the effect of the CPSA short fibers contained in the flexural strength of the glass-ionomer cement. METHOD: The 40 mass% short fibers of CPSA glass were added to the powder of commercial glass ionomer cement. Beam specimens of set cement (25 x 2 x 2 mm) were prepared for measuring the flexural strength (FS). The specimens of set commercial glass ionomer cement (GI) were used for comparison with glass ionomer cement with CPSA (FRGI). Half of all specimens were thermocycled in water for 60s of dwell time at 4 and 60 degrees C for 2500, 5000, and 10,000 cycles. The other specimens were stored in water at 37 degrees C as a control. The measurements were statistically analyzed using ANOVA. RESULTS: The initial FS of FRGI was 18.1 and that of GI was 7.7 MPa. After thermocycling, the mean FS ranged from 19.3 to 26.3 MPa and 9.8 to 11.1 MPa for FRGI and GI, respectively. After storage in water, the mean FS of FRGI and GI ranged from 20.4 to 25.9 MPa and 9.4 to 10.1 MPa, respectively. CONCLUSION: These findings suggested that glass-ionomer cement reinforced with CPSA short fibers maintains a higher strength than the conventional cement after aging. PMID- 11472812 TI - Polysaccharides in colon-specific drug delivery. AB - Natural polysaccharides are now extensively used for the development of solid dosage forms for delivery of drug to the colon. The rationale for the development of a polysaccharide based delivery system for colon is the presence of large amounts of polysaccharidases in the human colon as the colon is inhabited by a large number and variety of bacteria which secrete many enzymes e.g. beta-D glucosidase, beta-D-galactosidase, amylase, pectinase, xylanase, beta-D xylosidase, dextranase, etc. Various major approaches utilizing polysaccharides for colon-specific delivery are fermentable coating of the drug core, embedding of the drug in biodegradable matrix, formulation of drug-saccharide conjugate (prodrugs). A large number of polysaccharides have already been studied for their potential as colon-specific drug carrier systems, such as chitosan, pectin, chondroitin sulphate, cyclodextrin, dextrans, guar gum, inulin, amylose and locust bean gum. Recent efforts and approaches exploiting these polysaccharides in colon-specific drug delivery are discussed. PMID- 11472813 TI - Influence of storage on in-vitro release of ibuprofen from sugar coated tablets. AB - Studies performed on ibuprofen tablets (one brand of 400 mg, two brands of 200 mg sugar coated and one brand of film coated tablets) are reported. Tablets were subjected to conditions of 23 degrees C, 30 degrees C and 40 degrees C; at 75% RH and 96% RH for periods of up to 4 weeks. Tablets were stored in different ways- unpacked, packed in air-tight/moisture proof containers, packed in tablet vials and packed in two unit dose packs. Dissolution was carried out in pH 7.2 phosphate buffer using USP or FDA conditions for ibuprofen (Basket-150 rpm or Paddle-50 rpm) with sampling and UV analysis up to 90 or 120 min. Serious reduction in dissolution was noted for the 400 mg sugar coated tablets exposed to moisture. Mean % released at 30 mm (USP conditions) was as low as 1% and, for these tablets, dissolution continued to proceed extremely slowly for the full dissolution period. The film coated tablets were not affected. The tablet vials and unit dose packs showed some protection. Investigation showed not only a change in the subcoat properties (which did not break down easily) but also in the tablet core, which became hard and non-disintegrating. PMID- 11472814 TI - Evaluation of model solvent systems for assessing the accumulation of container extractables in drug formulations. AB - The interaction between a medical device and a pharmaceutical solution it contacts may dictate solution safety and/or efficacy. Of specific concern is the ability of device components to leach into the contacting solution. As pharmaceutical solutions containing surfactants, co-solvents and solubilizing agents become more common, method's for assessing the extent of leaching are needed. In this manuscript, a model is developed which relates a solution's polarity to its ability to interact with a plastic material. The validity of the developed model is examined via direct analysis of several pharmaceutically relevant solutions. PMID- 11472815 TI - Retention and transit of intestinal mucoadhesive films in rat small intestine. AB - The retention and transit characteristics of intestinal mucoadhesive film systems have been studied after intraduodenal administration in rats. Small size four layered film preparations, 0.5x0.5 mm, were prepared, where the backing layer (45.1+/-2.9 microm thick) was made of a water-insoluble polymer, ethylcellulose (EC), the surface layer was made of enteric pH-sensitive polymers, Eudragit L100, S100 or HP-55 and the middle layer was made of cellulose membrane. The surface layer was attached to the middle layer with an adhesive layer composed of carboxyvinyl polymer (Hiviswako(R) 103). After administration of ten films to the duodenum, the rats were sacrificed hourly and the distribution of the films in the whole small intestine was directly observed after abdominal incision. The HP 55, Eudragit L100 and S100 film systems were found to adhere to the upper, middle and lower part of the small intestine after 1, 2 and 4 h, respectively, for 2-3 h. Direct inspection study suggests that intestinal mucoadhesive film system has functions of: (1) pH-dependent intestinal adhesion site specificity; (2) adhesion to the intestinal wall; and (3) retention in the small intestinal adhesion site for at least 2 h. Intestinal mucoadhesive film system has been suggested to be a targeting system for drugs to the gastrointestinal tract. PMID- 11472816 TI - Effect of cholesterol content in activation of the classical versus the alternative pathway of rat complement system induced by hydrogenated egg phosphatidylcholine-based liposomes. AB - Liposomes composed of hydrogenated egg phosphatidylcholine (HEPC) and cholesterol (CHOL) were found to activate the rat complement (C) system in a CHOL content dependent manner. Liposomes containing 22 or 33 mol% CHOL activated the C system in a Ca(2+)-dependent manner, suggesting that C activation occurred via the classical pathway. Liposomes containing 44 mol% CHOL activated the C system in a Ca(2+) independent manner, suggesting that C activation occurred via the alternative pathway. The CHOL content appeared to dictate the pathway by which the C system was activated. This C activation was inhibited by removal of serum component(s), which adsorb to the liposomes. Activation of the alternative pathway, induced by the liposomes, was reduced by the depletion of IgG and IgM, whereas the classical pathway activation was reduced by the depletion of IgG, but not IgM. In addition, the removal of adsorbed serum component(s) by treatment with 44 mol% CHOL-containing liposomes decreased serum IgG and IgM levels that adsorb to the same liposomes, whereas the removal of adsorbed serum component(s) by treatment with 22 mol% CHOL-containing liposomes only slightly decreased serum IgG levels, which adsorbs to the same liposomes. Collectively, both IgG and IgM, which are specifically adsorbed to the liposomes in a CHOL-content dependent manner, were responsible for C activation via the alternative pathway induced by the 44 mol% CHOL containing liposomes. IgG alone would be partially responsible for C activation via the classical pathway induced by 22 or 33 mol% CHOL containing liposomes. The discovery of this unique C-activating property of liposomes will be of value in attempts to decipher the underlying mechanism of C activation by providing a useful model membrane system. PMID- 11472817 TI - In vitro studies on penetration of terpenes from matrix-type transdermal systems through human skin. AB - Polyurethane matrices containing up to 39% of the terpenes eucalyptol, L limonene, D-limonene, dipentene or terpinolene were produced. Release of the terpenes directly to the acceptor fluid, as well as through isolated human epidermis and dermis, was studied. In the presence of dermis the penetration profiles were very similar to the release profiles, indicating that dermis does not present a barrier for penetration of terpenes. For all terpenes the penetration was slower in the presence of epidermis (K(p) was in the range 0.21 1.8x10(-3) cm/h). Release and penetration through the epidermis and dermis were fastest for dipenetene (mixture of D-limonene and L-limonene), being at least 3-4 times faster than for D-limonene and L-limonene. Large amounts of terpenes found in epidermis (approximately 1.5 mg/cm(2)) indicate that affinity of these compounds to the stratum corneum is very high. PMID- 11472818 TI - In vitro and in vivo evaluations of topically applied capsaicin and nonivamide from hydrogels. AB - The purpose of this study was designed to investigate the in vitro and in vivo skin absorption of capsaicin and nonivamide from hydrogels. Various commercialized creams of capsaicin were also compared with hydrogels. Both skin stripping technique and Mexameter were applied to evaluate the level of capsaicin and nonivamide retained in stratum corneum (SC) and skin erythema in vivo. The partition of drug between skin and the hydrogel matrix was considered to play an important role in the permeation process. The in vitro permeation of capsaicin from hydrogels depends on the physicochemical nature and the concentration of the polymer used. The incorporation of nonionic Pluronic F-127 polymer into hydrogels resulted in a retarded release of capsaicin. On the other hand, the in vitro capsaicin permeation showed higher levels in cationic chitosan and anionic carboxymethyl cellulose (CMC) hydrogels than cream bases. The permeation of nonivamide was retarded at the late stage of in vitro application. The inter subject variation was more significant in the in vivo study than in vitro skin permeation experiments. The cream induced in vivo skin erythema depending on the drug concentration, however, the dose-dependence was not observed in hydrogels. Nonivamide-treated skin showed stronger erythema than capsaicin-treated skin. The present study indicates that there is a moderate correlation between in vitro skin permeation and in vivo erythema responses of topically applied capsaicin and nonivamide. The correlation between drug amount in SC and skin erythema test in vivo was also observed. PMID- 11472819 TI - Improved nasal bioavailability of elcatonin by insoluble powder formulation. AB - The bioavailability of elcatonin (ECT) via the nasal route was investigated with a powder dosage form utilizing water-insoluble calcium carbonate (CaCO(3)) in comparison with the liquid dosage form. Total radioactivity and the radioactivity of intact [3H]ECT were measured to evaluate the nasal absorption in vivo and the nasal mucosal transport in vitro. The systemic bioavailability of both total radioactivity and intact [3H]ECT following intranasal administration of the powder formulation in rats was significantly greater than in the case of the liquid formulation. In contrast, similar permeability of ECT across excised rabbit nasal mucosa was seen for both formulations, and was close to that of [14C]inulin, suggesting that the ECT transport is predominantly paracellular in each case. However, the powder formulation significantly prolonged the residence time of [3H]ECT in the rat nasal cavity, compared with the liquid formulation. We conclude that the powder formulation utilizing CaCO(3) improves the nasal bioavailability by increasing the residence time of ECT in the nasal cavity and is likely to be effective in increasing systemic drug delivery. PMID- 11472820 TI - Comparison of two hard keratinous substrates submitted to the action of a keratinase using an experimental design. AB - The influence of temperature, pH, keratinase concentration, substrate concentration and incubation time on the soluble proteins released by a new keratinase from Doratomyces microsporus was studied with a second-order experimental design. Only 15 or 18 spectrophotometric analyses were required to determine the optimal experimental conditions for this keratinase on nail and hoof. This study was carried out by measuring, according to Smith's method, the concentration of soluble proteins released by the enzyme on two substrates: nails and sheep hooves. Results give optimum conditions for the keratinase to release the soluble proteins: pH 8.2, keratinase concentration 0.14% (weight of keratinase lyophilisate/final volume) and substrate concentration 5% (weight of nail powder/final volume) for nails; temperature 38.8 degrees C, pH 9, substrate concentration 5% (weight of hoof powder/final volume) and a 5 h 55 min incubation time for hooves. PMID- 11472821 TI - The study of drug release from microspheres adhered on pig vesical mucosa. AB - The object of our work is the preparation of a mucoadhesive drug delivery system intended for intravesical application. In the present work, microspheres with Eudragit RS matrix polymer and different mucoadhesive polymers, i.e. chitosan hydrochloride (Ch), sodium salt of carboxymethyl cellulose (CMC) and polycarbophil (PC) were prepared to evaluate their influence on the mucoadhesive properties of microspheres. Different parameters were determined and their influence on pipemidic acid release from microspheres adhered on intact and damaged pig vesical mucosa was evaluated: swelling of polymers, mucoadhesion strength of polymeric films and drug dissolution according to USP XXIV method. The dissolution rate from microspheres containing different mucoadhesive polymers decreases as follows: PC>Ch>CMC. PC swelled to the largest volume among all polymers and as a result the fastest release of the drug from PC microspheres was obtained. The release rate of pipemidic acid from microspheres adhered on intact mucosa followed the order PC>CMC>Ch. These results show that both drug dissolution and mucoadhesion strength strongly influence drug release from adhered microspheres. The slowest release from Ch microspheres could be interpreted by the largest mucoadhesion strength of Ch polymeric films. The release rate of pipemidic acid from microspheres adhered on damaged mucosa followed the order PC=Ch>CMC. The results obtained on pathologically changed mucosa model support the indication of the role of glycosaminoglycans and polymer charge in the mucoadhesion process on vesical mucosa. Analysis of release data shows that the drug dissolution profiles follow the Higuchi kinetics better than the release profiles from adhered microspheres and different kinetics might be a consequence of different release mechanisms. PMID- 11472822 TI - Application of fractal geometry to dissolution kinetic study of a sweetener excipient. AB - In the context of relationship study between dissolution kinetic and particle morphology using the fractal geometry tool, we use a commercially available quality of saccharin powder. The characterization of molecular feature and image analysis study allows us to conclude to the statistic self-similarity of particles of four sieved particles size fractions, permitting the fractal approach. Calculation of reactive fractal dimension is performed using two forms of mass transfer equation: -dQ/dt=kQ(D(R)/3)DeltaC and -dQ/dt=k'R(D(R)-3)DeltaC, with DeltaC=(C(f)/[lnC(s)/(C(s)-C(f))]). Based on comparison of the surface fractal dimension D(S) on the two values of reactive fractal dimension D(R), a dissolution mechanism can be drawn: the dissolution starts at the whole surface of particles and is further governed by digging into holes that involve inner mass of particles. S.E.M. observations confirm this hypothesis. The confrontation between the D(R) values provided by the two ways of determination is essential for a good prediction of the mechanism. PMID- 11472823 TI - A simple method for evaluating the mixing efficiency of a new type of pan coater. AB - The objective of this study was to investigate some important process parameters on the mixing behavior of a new coater type. The coater used in this study, the Bohle BLC pan coater, differs from other coaters in its high length to diameter ratio. The pan coater can be divided into two zones: drying and spraying zone. The temperature difference between two points in the pan (one in each zone) was used to explore the influence of some process parameters, i.e. the pan speed and the inclination of the rotation axis on the mixing behavior. In addition, the effect of the spray rate on the temperature difference was studied. The results of the current study demonstrated the possibility to characterize the mixing behavior in a pan coater by a simple temperature measurement. The temperature difference method resulted in a model, which was comparable to the model from a conventional mixture experiment. It was shown that the pan speed and the spray rate influenced the temperature difference and, consequently, the mixing efficiency. However, the inclination of the rotation axis did not show an important effect. PMID- 11472824 TI - Investigation of active substance release from poly(ethylene oxide) hydrogels. AB - The uptake and controlled release of model active substances from poly(ethylene oxide), (PEO), hydrogels synthesized by irradiation were investigated. For the characterization of network structure of PEO hydrogels, swelling properties in water and the number average molecular weight between crosslinks were determined. Salicylic acid, phthalic acid and resorcinol were used as model substances for their controlled release from PEO hydrogels. The effects of dose rate, total dose and chemical structure of active substance on the uptake and release have been studied. The active substance uptake capacity of hydrogels was found to be lowest for phthalic acid and highest for resorcinol in the gel system obtained by irradiation both at low and high dose rates. The release was lowest both in rate and in total amounts in hydrogels containing phthalic acid, more in those with salicylic acid and highest in those with resorcinol. The physical and chemical factors affecting the release of model compounds such as the network structure of hydrogels and hydrogen bond formation between the adsorbent and PEO chains were discussed. PMID- 11472825 TI - Chitosan nanoparticles: a new vehicle for the improvement of the delivery of drugs to the ocular surface. Application to cyclosporin A. AB - Present limitations in the management of extraocular diseases include the inability to provide long-term extraocular drug delivery without compromising intraocular structures and/or systemic drug exposure. In the present study, the potential of chitosan (CS) nanoparticles as a new vehicle for the improvement of the delivery of drugs to the ocular mucosa was investigated. Cyclosporin A (CyA) was chosen as a model compound because of its potential usefulness for the treatment of these local diseases. An ionic gelation technique was conveniently modified in order to produce CyA-loaded CS nanoparticles. These nanoparticles had a mean size of 293 nm, a zeta potential of +37 mV and high CyA association efficiency and loading (73 and 9%, respectively). In vitro release studies, performed under sink conditions, revealed a fast release during the first hour followed by a more gradual drug release during a 24-h period. In vivo experiments showed that, following topical instillation of CyA-loaded CS nanoparticles to rabbits, it was possible to achieve therapeutic concentrations in external ocular tissues (i.e., cornea and conjunctiva) during at least 48 h while maintaining negligible or undetectable CyA levels in inner ocular structures (i.e., iris/ciliary body and aqueous humour), blood and plasma. These levels were significantly higher than those obtained following instillation of a CS solution containing CyA and an aqueous CyA suspension. From these results, we can conclude that CS nanoparticles may represent an interesting vehicle in order to enhance the therapeutic index of clinically challenging drugs with potential application at extraocular level. PMID- 11472826 TI - Stabilization of supersaturated solutions of a lipophilic drug for dermal delivery. AB - The stability of supersaturated solutions of a model lipophilic drug (LAP, a lavendustin derivative) in propylene glycol-water mixtures prepared using the method of mixed cosolvents was investigated. The solutions had a fixed degree of saturation (DS=4), but contained different ratios of propylene glycol-water. The absolute concentrations of LAP in these solutions varied by approximately a factor of 40, but the solutions at lower concentrations were no more stable than the more concentrated solutions. This shows that stability is primarily a question of the degree of saturation and not of the absolute drug concentration. Solutions of up to 5 degrees of saturation in 7:3 propylene glycol-water mixture were stable when stored for several hours; those at higher degrees of saturation recrystallized immediately. When the solutions were stirred, recrystallization occurred more rapidly. The influence of various polymeric additives on the stability of the supersaturated solutions showed that only sodium carboxymethyl cellulose had a stabilizing effect; however, the solution was very viscous and it is not clear whether the stabilizing effect was due to this high viscosity or to a specific interaction between drug and polymer. PMID- 11472827 TI - The formulation of an effective topical antibacterial product containing Ocimum gratissimum leaf essential oil. AB - The antibacterial potential of Ocimum gratissimum essential oil was explored. Liquid and semisolid formulations of the oil were designed in a variety of bases for topical antiseptic medication. The products were evaluated by agar diffusion assay against type strains and clinical isolates from boil, wound and pimples. Remarkable antibacterial effects, higher than those of commercial antiseptic products, were demonstrated at 2% Ocimum oil concentration in some bases. The properties of base into which the oil was incorporated affected its activity. It was more effective in hydrophilic bases than in lipophilic bases. Solubilization and microemulsification grossly reduced its activity. PMID- 11472828 TI - Quaternary ammonium palmitoyl glycol chitosan--a new polysoap for drug delivery. AB - A new polysoap, quaternary ammonium palmitoyl glycol chitosan (GCPQ, M(w)=178,000 g mole(-1)) with drug solubilising potential has been synthesised and characterised. In solution hydrophobic domains of GCPQ polymeric micelles were identified by the hypsochromic shift in the lambda(max) of methyl orange and by the increase in the ratio of the fluorescence emission intensity of the third and first pyrene vibronic peaks (I(3)/I(1)). At high aqueous concentrations (>10 mg ml(-1)) GCPQ presents as a gel which solubilises pyrene (2.5 mM, normal solubility in water approximately 2 microM) on probe sonication. Dilution of the gel to a liquid solution of polymeric micelles (< or =3.75 mg ml(-1) of GCPQ), results in the observation of fluorescent pyrene excimers (excited dimers) and a high excimer to monomer fluorescence ratio (I(E)/I(M)). However, attempts to solubilise pyrene at a concentration of 2.5 mM in a liquid solution of GCPQ (3.75 mg ml(-1)) results in a reduced I(E)/I(M) value and pyrene precipitation. Viscometry measurements show a more compact conformation for the polymer solubilising pyrene than the polymer alone. The polymer is non-haemolytic when present as the liquid solution and relatively non cytotoxic. In conclusion, a new biocompatible polysoap (potential drug solubiliser) is described which forms hydrophobic domains in solution and shows hysteresis in its solubilisation of pyrene. PMID- 11472829 TI - The prevention of cyclosporin A adsorption to Transwell surfaces by human plasma. AB - Prevention of cyclosporin A (CsA) adsorption onto the inner surface of Transwell during transport experiments, by the addition of human plasma to the receiver compartment (basolateral side), was investigated. The addition of plasma to a level of 50% (v/v) of the transport medium led to a reduction in the adsorption of CsA (0.1 microM) down to a level of 5%. As a result, the apical to basolateral flux of CsA across the Caco-2 cell monolayer in the presence of 50% (v/v) plasma was estimated to be 2.7-fold higher than that obtained in the absence of plasma. Thus, the adsorption problem can be overcome simply by the addition of an appropriate volume of human plasma to the transport medium. This method appears to be applicable to the routine estimation of CsA flux across epithelial cell monolayers using Transwell. PMID- 11472831 TI - A possible meiotic function of the peculiar patterns of gene expression in mammalian spermatogenic cells. AB - This review focuses on the striking differences in the patterns of transcription and translation in somatic and spermatogenic cells in mammals. In early haploid cells, mRNA translation evidently functions to restrict the synthesis of certain proteins, notably protamines, to transcriptionally inert late haploid cells. However, this does not explain why a substantial proportion of virtually all mRNA species are sequestered in translationally inactive free-messenger ribonucleoprotein particles (free-mRNPs) in meiotic cells, since most mRNAs undergo little or no increase in translational activity in transcriptionally active early haploid cells. In addition, most mRNAs in meiotic cells appear to be overexpressed because they are never fully loaded on polysomes and the levels of the corresponding protein are often much lower than the mRNA and are sometimes undetectable. A large number of genes are expressed at grossly higher levels in meiotic and/or early haploid spermatogenic cells than in somatic cells, yet they too are translated inefficiently. Many genes utilize alternative promoters in somatic and spermatogenic cells. Some of the resulting spermatogenic cell-altered transcripts (SCATs) encode proteins with novel functions, while others contain features in their 5'-UTRs, secondary structure or upstream reading frames, that are predicted to inhibit translation. This review proposes that the transcriptional machinery is modified to provide access to specific DNA sequences during meiosis, which leads to mRNA overexpression and creates a need for translational fine-tuning to prevent deleterious consequences of overproducing proteins. PMID- 11472832 TI - The function of leak and kuzbanian during growth cone and cell migration. AB - Axonal growth cones require an evolutionary conserved repulsive guidance system to ensure proper crossing of the CNS midline. In Drosophila, the Slit protein is a repulsive signal secreted by the midline glial cells. It binds to the Roundabout receptors, which are expressed on CNS axons in the longitudinal tracts but not in the commissural tracts. Here we present an analysis of the genes leak and kuzbanian and show that both genes are involved in the repulsive guidance system operating at the CNS midline. Mutations in leak, which encodes the Roundabout-2 Slit receptor, were first recovered by Nusslein-Volhard and co workers based on defects in the larval cuticle. Analysis of the head phenotype suggests that slit may be able to act as an attractive guidance cue while directing the movements of the dorsal ectodermal cell sheath. kuzbanian also regulates midline crossing of CNS axons. It encodes a metalloprotease of the ADAM family and genetically interacts with slit. Expression of a dominant negative Kuzbanian protein in the CNS midline cells results in an abnormal midline crossing of axons and prevents the clearance of the Roundabout receptor from commissural axons. Our analyses support a model in which Kuzbanian mediates the proteolytic activation of the Slit/Roundabout receptor complex. PMID- 11472833 TI - The foot formation stimulating peptide pedibin is also involved in patterning of the head in hydra. AB - Pedibin, a peptide of 21 amino acids, has been shown to stimulate foot formation in hydra, one of the simplest metazoan animals. The data presented here show that pedibin is synthesized as a precursor of 49 amino acids. A putative cleavage site precedes the peptide as purified from hydra tissue. The precursor, like pedibin, accelerates foot regeneration. Pedibin transcripts are concentrated in the foot region of hydra as expected, but are also present in the head region accumulating in the tentacle bases. The early appearance of pedibin transcripts during phases of cell fate specification like budding and regeneration implies that in hydra, pedibin plays an important role in patterning processes of foot and head. This is confirmed by the finding that pedibin also stimulates bud outgrowth. PMID- 11472834 TI - Top-DER- and Dpp-dependent requirements for the Drosophila fos/kayak gene in follicular epithelium morphogenesis. AB - The Drosophila fos (Dfos)/kayak gene has been previously identified as a key regulator of epithelial cell morphogenesis during dorsal closure of the embryo and fusion of the adult thorax. We show here that it is also required for two morphogenetic movements of the follicular epithelium during oogenesis. Firstly, it is necessary for the proper posteriorward migration of main body follicle cells during stage 9. Secondly, it controls, from stage 11 onwards, the morphogenetic reorganization of the follicle cells that are committed to secrete the respiratory appendages. We demonstrate that DER pathway activation and a critical level of Dpp/TGFbeta signalling are required to pattern a high level of transcription of Dfos at the anterior and dorsal edges of the two groups of cells that will give rise to the respiratory appendages. In addition, we provide evidence that, within the dorsal-anterior territory, the level of paracrine Dpp/TGFbeta signalling controls the commitment of follicle cells towards either an operculum or an appendage secretion fate. Finally, we show that Dfos is required in follicle cells for the dumping of the nurse cell cytoplasm into the oocyte and the subsequent apoptosis of nurse cells. This suggests that in somatic follicle cells, Dfos controls the expression of one or several factors that are necessary for these processes in underlying germinal nurse cells. PMID- 11472835 TI - Antagonistic regulation of convergent extension movements in Xenopus by Wnt/beta catenin and Wnt/Ca2+ signaling. AB - Convergent extension movements are the main driving force of Xenopus gastrulation. A fine-tuned regulation of cadherin-mediated cell-cell adhesion is thought to be required for this process. Members of the Wnt family of extracellular glycoproteins have been shown to modulate cadherin-mediated cell cell adhesion, convergent extension movements, and cell differentiation. Here we show that endogenous Wnt/beta-catenin signaling activity is essential for convergent extension movements due to its effect on gene expression rather than on cadherins. Our data also suggest that XLEF-1 rather than XTCF-3 is required for convergent extension movements and that XLEF-1 functions in this context in the Wnt/beta-catenin pathway to regulate Xnr-3. In contrast, activation of the Wnt/Ca2+ pathway blocks convergent extension movements, with potential regulation of the Wnt/beta-catenin pathway at two different levels. PKC, activated by the Wnt/Ca2+ pathway, blocks the Wnt/beta-catenin pathway upstream of beta-catenin and phosphorylates Dishevelled. CamKII, also activated by the Wnt/Ca2+ pathway, inhibits the Wnt/beta-catenin signaling cascade downstream of beta-catenin. Thus, an opposing cross-talk of two distinct Wnt signaling cascades regulates convergent extension movements in Xenopus. PMID- 11472836 TI - Impaired ossification in mice lacking the transcription factor Sp3. AB - Sp3 is a ubiquitously expressed member of the Sp family of transcription factors. Recently, the mouse Sp3 gene has been disrupted by homologous recombination. Sp3 null mice die immediately after birth due to respiratory failure. In addition, these mice show a pronounced defect in late tooth formation. Here we show that Sp3 is also required for proper skeletal ossification. Both endochondral and intramembranous ossification are impaired in E18.5 Sp3-/- embryos. The delay in ossification is reflected by reduced expression of the osteoblast-specific marker gene osteocalcin. The transcription factor - core binding factor 1 (Cbfa1)--that is essential for bone formation, however, is expressed at normal levels. In vitro differentiation studies using Sp3-/- ES cells further support the conclusion that Sp3 is needed for correct bone formation. The capacity of Sp3-/- cells to undergo osteogenic differentiation in vitro is reduced and osteocalcin expression is significantly diminished. Our studies establish Sp3 as an essential transcription factor for late bone development. PMID- 11472837 TI - The human chordin gene encodes several differentially expressed spliced variants with distinct BMP opposing activities. AB - During early embryogenesis of both vertebrates and invertebrates, antagonism between bone morphogenetic proteins (BMPs) and several unrelated secreted factors including Chordin (Chd) is a general mechanism by which the dorso-ventral axis is established. High affinity binding of Chd sequesters the BMP ligands in the extracellular space, preventing interactions with their membrane receptors. Another level of regulation consists in processing of vertebrate Chd or its Drosophila counterpart Sog by astacine metalloproteases like Xolloid-BMP 1/Tolloid, respectively, which releases an active BMP. Recently, it was shown that cleavage of Sog by Tolloid could generate novel BMP inhibitory activity and that sog is also capable of stimulation of BMP activity in a tolloid-dependant way. Activity and/or cleavage of Chd/Sog are influenced by other secreted factors like twisted gastrulation. In this study, we have cloned cDNAs of the human chordin gene (CHRD) and characterized alternative splice variants that code for C truncated forms of the protein. We have found that CHRD is expressed in fetal as well as in adult tissues with relatively high levels in liver, cerebellum and female genital tract, suggesting functions in late embryogenesis and adult physiology. We also show that spliced variants are present with specific patterns in various tissues. When tested in an axis-duplication assay in Xenopus, we find that these variants can antagonize BMP activity. Altogether, these results suggest that, in addition to processing by metalloproteases, alternative splicing (AS) is another mechanism by which sub-products of CHRD can be generated to influence BMP activity in different developmental and physiological situations. PMID- 11472838 TI - Tissue specific regulation of VEGF expression during bone development requires Cbfa1/Runx2. AB - Vascular endothelial growth factor (VEGF) is a critical regulator of angiogenesis during development, but little is known about the factors that control its expression. We provide the first example of tissue specific loss of VEGF expression as a result of targeting a single gene, Cbfa1/Runx2. During endochondral bone formation, invasion of blood vessels into cartilage is associated with upregulation of VEGF in hypertrophic chondrocytes and increased expression of VEGF receptors in the perichondrium. This upregulation is lacking in Cbfa1 deficient mice, and cartilage angiogenesis does not occur. Finally, over expression of Cbfa1 in fibroblasts induces an increase in their VEGF mRNA level and protein production by stimulating VEGF transcription. The results demonstrate that Cbfa1 is a necessary component of a tissue specific genetic program that regulates VEGF during endochondral bone formation. PMID- 11472839 TI - Comparative biological responses to human Sonic, Indian, and Desert hedgehog. AB - A comprehensive comparison of Sonic (Shh), Indian (Ihh), and Desert (Dhh) hedgehog biological activities has not previously been undertaken. To test whether the three higher vertebrate Hh proteins have distinct biological properties, we compared recombinant forms of the N-terminal domains of human Shh, Ihh, and Dhh in a variety of cell-based and tissue explant assays in which their activities could be assessed at a range of concentrations. While we observed that the proteins were similar in their affinities for the Hh-binding proteins; Patched (Ptc) and Hedgehog-interacting protein (Hip), and were equipotent in their ability to induce Islet-1 in chick neural plate explant; there were dramatic differences in their potencies in several other assays. Most dramatic were the Hh-dependent responses of C3H10T1/2 cells, where relative potencies ranged from 80nM for Shh, to 500nM for Ihh, to >5microM for Dhh. Similar trends in potency were seen in the ability of the three Hh proteins to induce differentiation of chondrocytes in embryonic mouse limbs, and to induce the expression of nodal in the lateral plate mesoderm of early chick embryos. However, in a chick embryo digit duplication assay used to measure polarizing activity, Ihh was the least active, and Dhh was almost as potent as Shh. These findings suggest that a mechanism for fine-tuning the biological actions of Shh, Ihh, and Dhh, exists beyond the simple temporal and spatial control of their expression domains within the developing and adult organism. PMID- 11472840 TI - Graded expression patterns of ephrin-As in the superior colliculus after lesion of the adult mouse optic nerve. AB - The idea has been put forward that molecules and mechanisms acting during development are re-used during regeneration in the adult, for example in response to traumatic injury in order to re-establish the functional integrity of neuronal circuits. Members of the Eph family of receptor tyrosine kinases and their 'ligands', the ephrins, play a prominent role during development of the retinocollicular projection in rodents, where EphA receptors and ephrin-As are expressed in gradients in both the retina and the superior colliculi (SC). We were interested in investigating whether EphA family members are also expressed or re-expressed in the adult after optic nerve lesion, since the presence of axon guidance information is an important prerequisite for a topographically appropriate re-connection by retinal ganglion cell (RGC) axons. This analysis was encouraged by results showing that RGC axons do not exert guidance preferences in response to membranes from adult unlesioned SC, but in response to membranes from the adult deafferented SC. We found a graded expression pattern of ephrin-As in the SC both before and after deafferentation, which was remarkably similar to those found during development. EphA receptor levels were reduced in the SC after deafferentation and the expression patterns of the EphB family were not changed. In particular, the presence of a graded ephrin-A expression in the deafferented SC suggests that - if robust regeneration of RGC axons can be achieved - topographic guidance information as a likely requirement for a functionally successful re-establishment of the retinocollicular projection is available. PMID- 11472841 TI - Proteolipid protein 2 mRNA is expressed in the rabbit embryo during gastrulation. AB - Differential display technology applied to rabbit blastocysts identified an mRNA that encodes a motif similar to that of the proteolipid protein PLP2/A4 of man, mouse and sheep. The open reading frame (456bp) has 88% amino acid identity to human PLP2/A4. The gene is maximally expressed at the beginning of gastrulation: in situ hybridizations exhibited a sickle-shaped area of labelling at the posterior pole of day 7 post-coitum embryos, which appeared at day 6.5 and decreased in size up to day 8. Weaker labelling was found in the extraembryonic mesoderm, in the anterior part of the primitive streak and in the trophoblast. Time and site of gene expression coincide with emerging morphogenetic activities at the posterior pole of the embryo at the beginning of gastrulation. PMID- 11472842 TI - Selective expression of angiopoietin 1 and 2 in mesenchymal cells surrounding veins and arteries of the avian embryo. AB - The expression of angiopoietin1 and 2 (ang1 and 2) and their receptor tie-2 was studied in avian embryos using in situ hybridization. Ang1 was transcribed in the mesenchymal cells surrounding venous endothelium expressing tie-2. By contrast, ang2 was transcribed around arteries in which the endothelium down-regulated tie 2 mRNA. The aorta and large arteries of the heart outflow tract were never surrounded by ang2 positive cells and maintained tie-2 expression. PMID- 11472843 TI - The Drosophila gene twister, an orthologue of the yeast helicase SKI2, is differentially expressed during development. AB - We have identified and characterized a Drosophila orthologue of SKI2, which, in Saccharomyces cerevisiae, is one of the key components in the cytoplasmic 3'-5' decay of mRNA. The Drosophila orthologue (twister, tst), is expressed as two transcripts which differ in the lengths of their 3'-UTRs, with the smaller transcript being particularly abundant in 0-2 h embryos and the larger transcript reaching its highest levels in 6-8 h embryos. TST protein is expressed in two forms which are differentially expressed in adult tissues and throughout development. Differential expression of TST may modulate activity of the mRNA turnover pathway and could have a major impact on the expression of target RNAs. PMID- 11472844 TI - Shh and Ptc are associated with taste bud maintenance in the adult mouse. AB - In mammals, taste receptor cells are organized into taste buds on tongue. Taste buds are trophically maintained by taste neurons and under continuous renewal, even in adults. We found that the receptor for Sonic hedgehog (Shh), Patched1 (Ptc), was expressed around taste buds where cells were proliferating, and that Shh was expressed within basal cells of taste buds. Denervation caused the loss of Shh and Ptc expression before the degeneration of taste buds. PMID- 11472845 TI - Expression of vinexin alpha in the dorsal half of the eye and in the cardiac outflow tract and atrioventricular canal. AB - Vinexin, a recently identified cytoskeletal protein, contains three SH3 domains and plays important roles in regulation of cytoskeletal organization and signal transduction. Using whole-mount in situ hybridization, we showed here that expression of vinexin alpha, the longer vinexin transcript, is strictly regulated, although the shorter transcript, vinexin beta, is expressed almost ubiquitously during embryonic development in mice. Expression of vinexin alpha was limited to within part of the eye and heart in 10.5 dpc embryos. Analysis of cryosections of 10.5 dpc embryos showed that vinexin alpha was expressed in a dorsal half of the retinal pigment epithelium and in the outflow tract and atrioventricular canal of the heart. Furthermore, we also found that vinexin alpha was expressed in the gonad and in a ventral part of the pons of 12.5 dpc embryos. These results indicated that the expression of vinexin alpha is strictly regulated in a temporally and spatially restricted manner. PMID- 11472846 TI - Meics, a novel zinc-finger protein which relocates from nuclei to the central meiotic spindle during Drosophila spermatogenesis. AB - A Drosophila gene encoding a novel zinc-finger protein, Meics, was cloned using a monoclonal antibody. The predicted amino acid sequence contains 12 zinc-finger motifs of the C2H2-type. During spermatogenesis, Meics distributes intranuclearly at pre- and post-meiotic stages whereas it relocates to central-spindle microtubules at both meiotic divisions. PMID- 11472847 TI - Irx1 and Irx2 expression in early lung development. AB - We describe a comparative lung expression analysis of the murine Irx1 and Irx2 genes. At embryonic day 8.5 (E8.5), the Irx1 and Irx2 expression starts in the foregut region, where the laryngo-tracheal groove will form. The expression is prominent in the lung epithelium during glandular development. It declines at the end of the canalicular phase. We further compare the Irx1 and Irx2 expression domains to Gli1, 2, 3 and Mash1. Their homologues in Drosophila melanogaster are known as regulative partners of the iroquois complex. The Irx and Gli genes are coexpressed in the developing lungs at the same time. Their transcripts are not localised in the same cells but adjacent to each other in either mesenchymal or epithelial structures. It is thought that the lung development is regulated by the mesenchymal/epithelial interactions. PMID- 11472848 TI - Expression pattern of Irx1 and Irx2 during mouse digit development. AB - Irx1 and Irx2 are members of the murine Iroquois homeobox (Irx) gene family. In this study, we describe the dynamic expression pattern of these genes during limb development with a focus on digit formation. We further present a comparative expression analysis with Gli genes (Gli1, Gli2, Gli3). Gli1, Gli2, and Gli3 were suggested for candidate regulators of the Irx genes. The expression was studied between E11.5 and E14.5 when the digits are being formed. Irx1 and Irx2 reproduce the developmental program of the digits in time and space and the Irx1 provides an early and excellent marker for this process. Our analysis also indicates that the expression of Irx1, Gli1 and Irx2, Gli2 are relative to each other. In contrast, Gli3 exhibits a different expression pattern. PMID- 11472849 TI - Expression patterns of the netrin receptor UNC5H1 among developing motor neurons in the embryonic rat hindbrain. AB - The axon guidance molecule netrin-1 has been implicated in the midline repulsion of developing cranial motor axons. We have examined expression patterns of the netrin receptors UNC5H1 and DCC in embryonic rat hindbrains, in combination with labelling of developing motor neurons. We found that UNC5H1 expression colocalised with a number of cranial motor neuron subpopulations from embryonic day 11 (E11) to E14, while DCC was expressed by motor neurons at E12. PMID- 11472850 TI - Embryonic expression of Kruppel-like factor 6 in neural and non-neural tissues. AB - Mammalian Kruppel-like transcription factors include 12 zinc finger proteins (KLF1-12) that are involved in regulation of cell proliferation and differentiation during morphogenesis and development (Trends Biochem. Sci., 24 (1999) 236). Structural considerations have segregated KLF6 and KLF7 into a separate sub-group, whereas in situ hybridizations have revealed predominant expression of the mouse klf7 gene in the developing nervous system. We examined the embryonic pattern of mouse klf6 in order to assess whether close kinship between KLF6 and KLF7 reflects similar expression patterns of the genes. The results of the in situ hybridizations demonstrate that klf6 expression in the developing nervous system is more restricted than klf7. In contrast to klf7, we also identified several non-neural sites of strong klf6 expression; they include the developing hindgut, heart, lung, kidney, and autopod. PMID- 11472851 TI - Cloning and developmental expression of STAT5 in Xenopus laevis. AB - Transcription factors of the signal transducer and activator of transcription (STAT) family are required for cellular responses to multiple signalling molecules. After ligand binding-induced activation of cognate receptors, STAT proteins are phosphorylated, hetero- or homodimerize, and enter the nucleus. STAT dimers bind to specific DNA elements and alter the transcriptional activity of the signal-responsive genes. We report the cloning and developmental pattern of expression of XSTAT5, a Xenopus laevis member of the STAT family, closely related to the mammalian STAT5A and STAT5B. XSTAT5 is expressed maternally and zygotically. With the onset of neurulation, XSTAT5 RNA are clearly localized in the anterior neural plate and subsequently in the neural structures of the developing eye, the pineal gland and the cement gland anlage. At late tailbud stages, a faint expression is detected in a ventral location that might correspond to the ventral blood islands. PMID- 11472852 TI - Expression patterns of Slit and Robo family members during vertebrate limb development. AB - We report the cloning and expression during limb development of the chicken Slit1, Slit2, and Slit3 ligands, and Robo1 and Robo2 receptor genes. We also compare the expression patterns of Robo1 and Robo2 in developing chick and mouse hindlimbs. These genes are expressed in regions of muscle development, chrondrogenesis, and axon guidance. PMID- 11472853 TI - Dynamic expression patterns of the new protocadherin families CNRs and Pcdh-gamma during mouse odontogenesis: comparison with reelin expression. AB - Protocadherins are transmembrane glycoproteins belonging to the cadherin superfamily of molecules, which are involved in many biological processes such as cell adhesion, cytoskeletal organization and morphogenesis. Protocadherins generally exhibit only moderate adhesive activity and are highly expressed in the nervous system. Here, we report on the expression pattern of two novel families of protocadherins (CNRs and Pcdh-gamma) during rodent teeth development. Furthermore, we compare their expression with that of reelin, which is the potential ligand of CNRs. Throughout odontogenesis, CNRs, Pcdh-gamma and reelin show dynamic spatiotemporal expression patterns, which relate to both morphogenesis and cell differentiation events. PMID- 11472854 TI - Specific expression of the retinoic acid-synthesizing enzyme RALDH2 during mouse inner ear development. AB - Retinoid binding proteins and nuclear receptors are expressed in the developing mouse inner ear. Here, we report that the retinaldehyde dehydrogenase 2 (Raldh2) gene, whose product is involved in the enzymatic generation of retinoic acid (RA), exhibits a restricted expression pattern during mouse inner ear ontogenesis. The Raldh2 gene is first expressed at embryonic day (E) 10.5 in a V shaped medio-dorsal region of the otocyst outer epithelium, which evolves as two separate domains upon otocyst morphogenesis. At E14.5, Raldh2 is expressed in two areas of the utricle epithelium and specific regions of the saccule and cochlear mesenchyme. Later, Raldh2 transcripts are restricted to two cochlear areas, the stria vascularis and Reissner membrane. Raldh2 mesenchymal expression did not correlate with migrating neural crest-derived melanoblasts. These restricted expression domains may correspond to specific sites of RA synthesis during inner ear morphogenesis. PMID- 11472855 TI - Abdominal B-type Hox gene expression in Xenopus laevis. AB - We have studied the expression of the Abdominal B-type Hox genes in Xenopus embryos and tadpoles. The probes used represent all paralog groups and are designated Xhoxa9, Xhoxd9, Xhoxd10, Xhoxa11, Xhoxc12 and Xhoxa13 on the basis of comparison to other vertebrates. Three of these genes are novel while the others have previously been detected but expression patterns were only partially described. We find a typical nested pattern of expression in the main body axis, in both central nervous system and mesoderm. All the genes, except Xhoxc12, are also expressed in the mesenchyme of the large intestine and in the limb buds. PMID- 11472856 TI - Nato3 is an evolutionarily conserved bHLH transcription factor expressed in the CNS of Drosophila and mouse. AB - The evolutionarily conserved basic helix-loop-helix (bHLH) transcription factors play important roles during development. Here we report the identification of Nato3 (nephew of atonal fer3) orthologs in Drosophila, C. elegans, mouse, and man, all of which share a high degree of similarity within the bHLH domain. Expression analysis revealed Nato3 transcripts in the central nervous system of both fly and mouse embryos. In the fly, Dnato3 is highly expressed in 9-15h embryos in a few ventral nerve cord cells and a subset of neurons in the brain. In mouse, the MNato3 transcripts were detected from embryonic day 7 until 5 weeks postnatally, with highest levels in the midbrain, thalamus, hypothalamus, pons, and medulla oblongata. In contrast to the brain, expression in the spinal cord was limited to the embryonic stages. PMID- 11472857 TI - Cell- and stage-specific high-level expression of TBP-related factor 2 (TRF2) during mouse spermatogenesis. AB - Mice lacking the TBP-related factor 2 (TRF2) gene, which is highly expressed in the testis, have a severe defect in spermiogenesis. Here we show that the expression of TRF2 is both cell type- and stage-specific. TRF2 expression was first detected in the late pachytene spermatocytes at stage VIII and increased throughout the subsequent stages. After meiotic divisions, the TRF2 expression declined continuously in round spermatids during progression from stage I to stage V. This observation is consistent with an essential regulatory role of TRF2 in male germ cell differentiation during spermatogenesis. PMID- 11472858 TI - Pattern of expression of the tetraspanin Tspan-5 during brain development in the mouse. AB - Here, we report the pattern of expression of the tetraspanin mTspan-5 at various developmental stages. Expression was first seen at E10 and remained until adulthood, with increased levels between P0 and P5. mTspan-5 showed high expression in the cortical areas and Purkinje cells throughout development. Moreover, transcripts were also detected in the developing heart, forelimbs, and hindlimbs. PMID- 11472859 TI - High frequency repetitive transcranial magnetic stimulation (rTMS) of the left dorsolateral cortex: EEG topography during waking and subsequent sleep. AB - Repetitive transcranial magnetic stimulation (rTMS) is a novel research tool in neurology and psychiatry. It is currently being evaluated as a conceivable alternative to electroconvulsive therapy for the treatment of mood disorders. Eight healthy young (age range 21-25 years) right-handed men without sleep complaints participated in the study. Two sessions at a 1-week interval, each consisting of an adaptation night (sham stimulation) and an experimental night (rTMS in the left dorsolateral prefrontal cortex or sham stimulation; crossover design), were scheduled. In each subject, 40 trains of 2-s duration of rTMS (inter-train interval 28 s) were applied at a frequency of 20 Hz (i.e. 1600 pulses per session) and at an intensity of 90% of the motor threshold. Stimulations were scheduled 80 min before lights off. The waking EEG was recorded for 10-min intervals approximately 30 min prior to and after the 20-min stimulations, and polysomnographic recordings were obtained during the subsequent sleep episode (23.00-07.00 h). The power spectra of two referential derivations, as well as of bipolar derivations along the antero-posterior axis over the left and right hemispheres, were analyzed. rTMS induced a small reduction of sleep stage 1 (in min and percentage of total sleep time) over the whole night and a small enhancement of sleep stage 4 during the first non-REM sleep episode. Other sleep variables were not affected. rTMS of the left dorsolateral cortex did not alter the topography of EEG power spectra in waking following stimulation, in the all-night sleep EEG, or during the first non-REM sleep episode. Our results indicate that a single session of rTMS using parameters like those used in depression treatment protocols has no detectable side effects with respect to sleep in young healthy males. PMID- 11472860 TI - Estrogen use and brain metabolic change in older adults. A preliminary report. AB - Because estrogen may influence brain blood flow and metabolism in older adults, we used positron emission tomography to evaluate cerebral glucose metabolic change in post-menopausal women and men. Women estrogen users (n=4), women non users (n=8) and men (n=10) were scanned at baseline and two years later. Analyses focused on glucose metabolism in lateral temporal, inferior parietal and posterior cingulate brain regions, previously reported to decline in non-demented older persons. No metabolic differences in cerebral regions of interest were found among groups at baseline. At follow-up, women estrogen users showed significantly increased glucose metabolism in the lateral temporal region, whereas women non-users and men exhibited no significant metabolic change in this region. These findings suggest that estrogen use may protect against regional cerebral metabolic decline in postmenopausal women. PMID- 11472861 TI - Electrophysiological signs of reduced prefrontal response control in schizophrenic patients. AB - The prefrontal cortex is considered as a brain region important in the etiopathogenesis of schizophrenic disorders. Based on converging results from different research areas, the prefrontal cortex is regarded as the anatomical and functional representation of response control under physiological conditions. In previous studies, a robust electrophysiological marker for the investigation of response control in healthy control subjects was validated. This parameter was termed NoGo anteriorisation and consists of a more anterior peak of the event related potentials during the inhibition of a prepared motor response (NoGo condition within the Continuous Performance Test) than during its execution (Go condition). The present study investigated these brain electrical correlates of response control in 19 schizophrenic patients and 19 age- and sex-matched healthy subjects. Compared to control subjects, the event-related potentials in schizophrenic patients were located more anterior in the Go condition and, as a trend, more posterior in the NoGo condition. The NoGo anteriorisation was strongly reduced in the schizophrenic group. On a qualitative level, the NoGo anteriorisation was present in all control subjects, but not in eight of the 19 patients. The results were interpreted as an indication of a disturbed prefrontal response control in schizophrenic patients. Further studies will clarify whether this method may be useful as a global test of hypofrontality in different groups of chronic schizophrenias, or as a quantifiable measure of an affected response control system, especially in catatonic subgroups. PMID- 11472862 TI - Mapping cortical asymmetry and complexity patterns in normal children. AB - This study reports the first comprehensive three-dimensional (3D) maps of cortical patterns in children. Using a novel parametric mesh-based analytic technique applied to high-resolution T1-weighted MRI scans, we examined age (6-16 years) and gender differences in cortical complexity (the fractal dimension or complexity of sulcal/gyral convolutions) and asymmetry of 24 primary cortical sulci in normally developing children (N=24). Three-dimensional models of the cerebral cortex were extracted and major sulci mapped in stereotaxic space. Given the documented age-related changes in frontal lobe functions and several neuroimaging studies that have reported accompanying volumetric changes in these regions, we hypothesized that, with age, we would find continued modifications of the cerebrum in frontal cortex. We also predicted that phylogenetically older regions of the cerebrum, such as olfactory cortex, would be less variable in anatomic location across subjects and with age. Age-related increases in cortical complexity were found in both left and right inferior frontal and left superior frontal regions, possibly indicating an increase in secondary branching with age in these regions. Moreover, a significant increase in the length of the left inferior frontal sulcus and a posterior shifting of the left pre-central sulcus was associated with age. Three-dimensional asymmetry and anatomic variability maps revealed a significant left-greater-than-right asymmetry of the Sylvian fissures and superior temporal sulci, and increased variance in dorsolateral frontal and perisylvian areas relative to ventral regions of the cortex. These results suggest increases in cortical complexity and subtle modifications of sulcal topography of frontal lobe regions, likely reflecting ongoing processes such as myelination and synaptic remodeling that continue into the second decade of life. More studies in a larger sample set and/or longitudinal design are needed to address the issues of normal individual variation and sulcal development. PMID- 11472863 TI - Abnormal neuronal network in anorexia nervosa studied with I-123-IMP SPECT. AB - Single photon emission computed tomography was used to study 14 female patients with anorexia nervosa and 8 female normal comparison subjects. Automatic voxel based analysis of the images was carried out using statistical parametric mapping (SPM) software. Statistics across the entire brain were displayed as Z scores (threshold: P<0.05). Compared with the normal comparison subjects, the anorectic patients were characterized by hypoperfusion in the medial prefrontal cortex and the anterior cingulate gyrus, and hyperperfusion in the thalamus and the amygdala hippocampus complex. These results suggest that a dysfunction in neuronal circuitry may be related to anorexia nervosa. PMID- 11472864 TI - Tc-99m HMPAO brain perfusion SPECT in drug-free obsessive-compulsive patients without depression. AB - The aim of this study was to confirm prior results of brain-imaging studies on obsessive-compulsive disorder (OCD) in a sample of Turkish patients, as a cross cultural study. Tc-99m HMPAO brain perfusion SPECT imaging was performed in nine drug-free OCD patients without depression and six controls. The patients' Hamilton Depression Rating Scale scores were <16. The severity of obsessive compulsive symptoms was rated with the Yale-Brown Obsessive-Compulsive Rating Scale (YBOCS). Quantitative evaluation of regional cerebral blood flow revealed that right thalamus, left frontotemporal cortex and bilateral orbitofrontal cortex showed significant hyperperfusion in patients with OCD compared with controls. YBOCS scores did not show any correlation with hyperperfusion in regional cerebral blood flow in these areas. Results of this cross-cultural study may support orbitofrontal and thalamic dysfunction in OCD in a sample of Turkish patients. PMID- 11472865 TI - Adolescent patient confidentiality: whom are we kidding? PMID- 11472866 TI - Impact of parental consent and notification policies on the decisions of adolescents to be tested for HIV. AB - Restrictions have been placed on the ability of adolescents to obtain human immunodeficiency virus (HIV) testing independent of their parents. Although some states have given adolescents the right to consent to HIV testing independently, many states have remained silent on the issue or have compromised these rights by providing for parental consent or notification when adolescents seek testing. This article examines existing policies and explores whether policies that require adolescents to obtain parental consent, or that permit or mandate parental notification, may deter them from obtaining needed HIV testing. PMID- 11472867 TI - Treatment of chronic nightmares in adjudicated adolescent girls in a residential facility. AB - PURPOSE: To evaluate imagery rehearsal therapy for the treatment of chronic nightmares in a sample of adolescent girls. METHODS: Adolescent girls ranging in age from 13 to 18 years were recruited from the Wyoming Girls School in Sheridan, Wyoming (treatment group, n = 9; control group, n = 10). These girls had previously suffered a high prevalence of unwanted sexual experiences in childhood and adolescence, and thus many suffered from nightmares, sleep complaints, and posttraumatic stress symptoms. Imagery rehearsal therapy was provided in a 1-day (6-h) workshop. Imagery rehearsal consists of three steps, all of which are performed in the waking state: (a) select a nightmare, (b) "change the nightmare any way you wish," and (c) rehearse the images of the new version ("new dream") 5 to 20 min each day. Control participants received no intervention. RESULTS: At baseline, these girls had been suffering from nightmares, on average, for 4.5 years, and they reported experiencing 20 nightmares per month, which occurred at a frequency of at least one bad dream every other night. At 3 months, self reported, retrospectively assessed nightmare frequency measured in nights per month decreased 57% (p =.01, d = 1.4) and measured in nightmares per month decreased 71% (p =.01, d = 1.7) in the treatment group, compared with no significant changes in the control group. No significant changes were noted for sleep and posttraumatic stress disorder measures in either group. CONCLUSION: Imagery rehearsal therapy was an effective treatment option for chronic nightmares in this adjudicated adolescent population. PMID- 11472868 TI - Predictors of late-onset smoking and cessation over 10 years. AB - PURPOSE: To identify predictors of smoking onset and cessation between early (age 13 years) and late adolescence (age 18 years) and between late adolescence (age 18 years) and young adulthood (age 23 years). METHODS: We employed logistic regression to predict smoking initiation and cessation for an ethnically diverse sample of 3056 adolescents recruited from 30 West Coast schools in 1985 and observed from age 13 to age 23 years. Fifty-six percent of the sample was female. Predictors tapping sociodemographic characteristics, environmental influences, attitudes and beliefs about smoking, bonds with school, and problem behavior were measured at age 13 years (older teenager models) and at age 18 years (young adult models). RESULTS: Robust predictors of both initiation and cessation across the two developmental periods included doing poorly in middle/high school and prior smoking behavior. Predictors common to three of the four models included being young for one's grade cohort and intending to smoke in the next 6 months. Early deviant behavior and drinking fostered initiation among older teenagers, but problem behavior as an older teenager did not predict young adult initiation. Smokers who had few or no high school friends who smoked and felt able to resist prosmoking pressures at age 18 years were more likely to quit by age 23 years. Being female predicted initiation by age 18 years; being African-American, Hispanic, or Asian inhibited it. CONCLUSIONS: The strong association of prior smoking behavior and intentions with later smoking status among both adolescents and young adults underscores the importance of starting smoking prevention early and continuing it through high school. Such programs might also consider the greater vulnerability of females, youth who are young for their grade cohort, and those who are doing poorly in school. PMID- 11472869 TI - Dealing with daily hassles: smoking and African-American adolescent girls. AB - PURPOSE: To examine cigarette use and its relationship to daily life hassles in an urban sample of African-American adolescent girls. METHODS: A sample of 105 African-American adolescent girls (mean age of 15.45 years) derived from a larger cross-sectional research project titled "Female Adolescent Substance Experience Study" funded by the National Institute of Drug Abuse comprised the sample. The sample was divided into adolescents who had ever smoked in their lifetime and adolescents who had never smoked before. Student's t-tests were conducted to determine whether there were differences between these groups on demographic characteristics and the number of daily life hassles. Pearson product moment correlations were also conducted to examine the association between age of smoking initiation and number of hassles. RESULTS: Less than 50% of the teenagers had ever smoked cigarettes in their lifetime, and of those who had ever smoked, the average age of initiation was 12.55 years (SD = 2.63). Furthermore, girls who had ever smoked, in contrast to girls who had never smoked, had a significantly greater number of daily life hassles, in general, and within the school/academic and family/economic domains in particular. Age of smoking initiation was negatively related to the number of hassles, indicating that girls who started to smoke at a younger age reported more hassles. CONCLUSIONS: These findings are discussed in terms of developing an understanding of gender and ethnic-specific correlates of smoking that can be used to better delineate the developmental smoking trajectory of African-American girls. PMID- 11472870 TI - Impact of a school-based intervention on access to healthcare for underserved youth. AB - PURPOSE: To determine whether a multidimensional school-based intervention, which included physical and mental health services, increased adolescents' use of needed medical care and preventive care and decreased emergency room use. METHODS: A total of 2832 seventh- through twelfth-grade students in six public urban intervention schools and 2036 students in six demographically matched comparison schools completed a previously validated survey regarding health status and healthcare utilization in spring 1998 and 1999. Bivariate analyses examined the association between intervention status and Year 1/Year 2 outcomes. The multifaceted intervention included programs such as anger management groups, substance abuse prevention, tutoring, home visits, and enhanced school health services. Stepwise multivariate logistic models tested differences between the intervention and comparison groups across years, controlling for potential confounding variables [gender, age, race/ethnicity, maternal education, grade in school, school district (city or county), health status, and chronic health problems]. The interaction term for Group x Year was used to test the effect of the intervention. Multivariable modeling was also used to determine student factors independently associated with healthcare utilization. RESULTS: Respondents had a median age of 15 years, 56% were female, 51% were white, 42% were black, and 34% reported chronic health problems. In both years, over 45% of students in both groups reported not seeking medical care they believed they needed. The proportion with missed care in the intervention schools did not change, whereas the proportion with missed care in the comparison schools increased. Emergency room use decreased slightly in the intervention schools and increased slightly in the comparison schools between Year 1 and Year 2. There were no major changes in healthcare delivery in this area during the year, demonstrating the volatility of adolescents' perceived access to care. Among the student factors, health status, having a chronic condition, and being in a higher grade were independently associated with students' report of not seeking care they believed they needed. CONCLUSION: These results confirm that many adolescents have unmet healthcare needs. Those with poor health status are most likely to report underutilization and unmet needs. These findings underscore the need for comparison groups when evaluating interventions and suggest the need for better understanding of community level changes in perceived healthcare access and use. PMID- 11472871 TI - Reliability of the Problem Oriented Screening Instrument for Teenagers (POSIT) in adolescent medical practice. AB - PURPOSE: To determine the internal consistency and 1-week test-retest reliability of the Problem Oriented Screening Instrument for Teenagers (POSIT) among adolescent medical patients. METHODS: A research assistant administered the POSIT to a consecutive sample of 15- to 18-year-old patients arriving for routine medical care at a hospital-based adolescent medical practice. Each subject returned for a retest 1 week later. Internal consistency for each scale on test and retest was calculated using Cronbach alpha, and 1-week test-retest reliability by the intraclass correlation coefficient (r) and the kappa coefficient (kappa). RESULTS: The Substance Use/Abuse, Mental Health Status, Educational Status, and Aggressive Behavior/Delinquency scales had favorable alpha scores (>.70). Others, including Physical Health Status, had lower alpha scores. High intraclass correlation coefficients were found for all 10 POSIT scales (r =.72 to.88), although (r) was lower for males on two of the scales. Kappa coefficients for all scales indicated good reproducibility beyond chance (kappa =.42 to.73). CONCLUSIONS: This study provided supportive evidence for the reliability of the POSIT in primary care medical settings, although some POSIT scales could likely be improved. The 20- to 30-min administration time is most practical in settings that are dedicated to adolescent medicine, and computerized administration and scoring are needed. PMID- 11472872 TI - The Adolescent Health Review: a brief, multidimensional screening instrument. AB - PURPOSE: To develop a brief, multidimensional screening instrument for adolescents that addresses psychosocial domains critical to adolescent preventive health care services. METHODS: Secondary analyses were conducted on survey data obtained in 1995 from a school sample of 76,159 students in grades 9 and 12, as well as 893 adolescents from juvenile correctional facilities, 500 adolescents from chemical dependency treatment programs, and 575 adolescents from residential behavioral treatment programs. A comprehensive set of 300 survey items was used in a series of discriminant analyses to determine which items best distinguished males and females in each clinical sample from their counterparts in the school sample. RESULTS: The item selection for the Adolescent Health Review was guided both by empirical analyses and clinical judgment. The final screen is comprised of 33 demographic and clinical items that address a variety of psychosocial domains. The computerized, self-administered screen can be completed in about 3 minutes. The screen is scored automatically and produces an easy-to-read risk assessment profile. Because screening items were drawn from a large epidemiologic survey, normative profiles are available for each age and gender subgroup. CONCLUSIONS: A brief, empirically derived screening instrument, designed to address a range of adolescent risks, offers an opportunity for information gathering that otherwise might not be incorporated into routine clinic visits. PMID- 11472873 TI - The health and well-being of adolescents: a school-based population study of the self-report Child Health Questionnaire. AB - PURPOSE: To evaluate a new generic measure of adolescent health status, the self report version of the Child Health Questionnaire (CHQ), and provide population based data. Furthermore, we aimed to examine the impact of common adolescent illness and health concerns on their health and well-being. METHODS: A stratified, two-stage, random cluster sampling design was used to obtain a cross sectional sample of subjects through schools. A written questionnaire included the 80-item 12-scale self-report CHQ and items measuring health concerns, illnesses/health conditions, and sociodemographics. RESULTS: A total of 2361 adolescents participated (response rate of 70%). Reliability was high: Tests of internal consistency and discriminant validity reported 90% of item-scale correlations >.4; all scales had Cronbach alpha coefficients >.7. Adolescents with illnesses/conditions or health concerns reported lower scores and larger differences for content-related scales, supporting content and construct validity. Statistically significant age and gender trends were observed for Mental Health, Self-Esteem, General Health, and Family Cohesion scales. Health status worsened as health concerns increased (X(2) linear trend, p =.00) with deterioration in health of 5-20% on all scales for emotional health concerns (40% of sample). CONCLUSIONS: The self-report CHQ is a reliable and seemingly valid measure of health and well-being for adolescent health research, although additional measures may be required where scales have high ceiling values. The significantly lower scores reported by adolescents with illness and/or health concerns lend support to the use of standardized health measures and longitudinal research to further examine the impact of adolescent comorbidities and their causal determinants. PMID- 11472874 TI - The long-term response to levodopa in dopa-responsive dystonia. AB - We report the long-term response to levodopa in 20 patients with dopa-responsive dystonia (DRD). We found an inverse correlation between the daily dose of levodopa and duration of treatment (r=-0.59, P<0.01). Mild dyskinesias were present in 20% of our patients. Dyskinetic patients were on a higher dose of levodopa than non-dyskinetics. Dyskinesias responded to a reduction in levodopa, with no deterioration in motor function. We propose that the dopamine turnover might decrease with time, which would lead to a decrease in the requirement for levodopa and the occurrence of dyskinesias late in the course of DRD. PMID- 11472875 TI - Alpha-synuclein expression is up-regulated in NTera2 cells during neuronal differentiation but unaffected by exposure to cytokines and neurotrophic factors. AB - Increasing evidence has indicated that proinflammatory cytokines such as TNF alpha and IL-1beta, produced by activated microglia and astrocytes, play a key role in progressive degeneration of the nigrostriatal dopaminergic neurons in Parkinson's disease (PD). Since alpha-synuclein is a major component of Lewy bodies in PD brains, we studied the constitutive and cytokine/neurotrophic factor regulated expression of alpha-synuclein in cultured human neurons by Northern blot and Western blot analyses. The constitutive expression of alpha-synuclein mRNA was identified in a variety of human neural and non-neural cell lines. The levels of alpha-synuclein expression were elevated markedly in NTera2 teratocarcinoma cells following retinoic acid-induced neuronal differentiation, accompanied with an increased expression of synphilin-1, while they were unaltered in NTera2-derived differentiated neurons by exposure to TNF-alpha, IL 1beta, BDNF or GDNF. These results indicate that alpha-synuclein expression in human neurons is up-regulated during differentiation, but is unaffected by a panel of cytokines and neurotrophic factors which are supposed to be involved in the nigral neuronal death and survival. PMID- 11472876 TI - The parkinsonian neurotoxin, MPP+ induces phosphorylated c-Jun in dopaminergric neurons of mesencephalic cultures. AB - The stress-activated protein kinase (SAPK) cascade serves a critical role in the apoptotic death of neuronal cells in response to a variety of cellular stresses. Recent in vitro and in vivo evidence has directly implicated this kinase in the death of dopaminergic nigral neurons in the MPTP model of Parkinson's disease (PD). To assess the involvement of c-Jun, a key transcription factor substrate of SAPK (also known as c-Jun N-terminal kinase, or JNK) in the MPTP-induced death of dopaminergic nigral neurons, we determined the ability of MPP+, the active toxin metabolite of MPTP, to induce the phosphorylated form of c-Jun in dopaminergic neurons in nigral (ventral mesencephalon) cultures. At a dose of MPP+ that specifically induces apoptotic changes in nuclear morphology in tyrosine hydroxylase-positive (dopaminergic) cells in these cultures, MPP+ induces nuclear phospho-c-Jun immunoreactivity (IR). The peak induction of phospho-c-Jun IR was observed 16h after beginning MPP+ exposure, and preceded the maximal induction of apoptotic nuclear changes by approximately 8h. These data support an important role for the SAPK/JNK pathway including its c-Jun transcriptional target in the apoptotic death of dopaminergic nigral neurons in the MPTP model of PD. PMID- 11472877 TI - CSF-neurofilament and levodopa tests combined with discriminant analysis may contribute to the differential diagnosis of Parkinsonian syndromes. AB - The differentiation between Parkinson's disease (PD), progressive supranuclear palsy (PSP) and multiple system atrophy (MSA) is important for prognostic and therapeutic purposes. In order to evaluate the diagnostic capability of two tests reflecting these items, patients fulfilling strict clinical criteria for PD (n=35), MSA (n=36) and PSP (n=14), were consecutively included. An analysis of neurofilament protein (NFL), a marker of axonal degeneration in the cerebrospinal fluid (CSF) and a levodopa test, recorded with optoelectronic technique were performed. Using discriminant analyses, the test's abilities to predict the clinical PD or non-PD (MSA and PSP) diagnoses were compared. Whereas the CSF-NFL and levodopa tests predicted 79 and 85% correct diagnoses respectively, the combined test predicted 90% correct diagnoses. We conclude that the CSF-NFL and levodopa tests provide detailed information of clinical variables on which the clinical diagnostic criteria are based. As they are pathologically unrelated, the diagnostic precision increases compared to clinical diagnoses when they are combined. PMID- 11472878 TI - Pharmacological involvement of the calcium channel blocker flunarizine in dopamine transmission at the striatum. AB - Single intrastriatal microinjections of 25, 50 and 100nmol/microl of flunarizine in normal rats produced a dose-dependent turning behavior toward the injected side when they were challenged with apomorphine (1mg/kg, s.c). This effect was seen at 1, 3 and 7 days following administration of the high dose of flunarizine, but had subsided by 24h after administration of the intermediate dose; the low dose was ineffective. However, intrastriatal injection of the high dose of flunarizine resulted in a local lesion and thereafter this dose was not used. A similar dose-response relationship was determined for nifedipine, an L-type calcium channel antagonist. Injection of this antagonist did not result in apomorphine-elicited rotational behavior, reflecting its lack of antidopaminergic action. Intrastriatal injections of haloperidol (5microg/microl), an antagonist of dopamine D(2) receptors, or the sodium channel blocker lidocaine (40microg/microl), were given in order to compare their effects to those observed with flunarizine. Intracerebral injection of haloperidol produced ipsilateral turning in response to systemic administration of apomorphine given 60min after. The same response was obtained with the injection of apomorphine 10min after the injection of intracerebral lidocaine. This effect was no longer apparent 24h after the microinjection of haloperidol and 60min after the injection of lidocaine. In rats rendered hemiparkinsionian by lesioning the nigrostriatal pathway with 6OHDA, intrastriatal microinjection of flunarizine (50nmol/microl) significantly reduced apomorphine (0.2mg/kg, s.c.)-elicited turning behavior towards the non-lesioned side. These results suggest an antidopaminergic effect of flunarizine mediated by antagonistic action of post-synaptic striatal dopamine receptors. However, an action of the drug on sodium channels may not be ruled out. These studies offer additional supporting evidence for the induction or aggravation of extrapyramidal side-effects in patients receiving flunarizine. PMID- 11472879 TI - Longitudinal evaluation of economic and physical impact of Parkinson's disease. AB - The cost of parkinsonism and Parkinson's disease (PD) is largely unknown although clinical experience suggests that the impact of this disease is substantial. Longitudinal data is presented for health status, disease symptoms, functional status, and financial costs for 70 participants with PD or parkinsonism. The sample was dichotomized into those rating their health as excellent, good, or very good ('good health') and those rating their health as fair or poor ('poor health'). The 'poor health' group were significantly more disabled at baseline. Symptoms increased between year 1 and 3 with greatest increases in fatigue, pain, and depression for the 'good health' group. At year 1, total direct cost/capita was about dollars 5000/year for both groups; indirect costs were dollars 5000 for the 'good health' group and dollars 15,000/year for the 'poor health' group. By year 3, total expenditures increased over 25% for the 'good health' group and nearly doubled for the 'poor health' group, while percent costs that were compensated declined for groups. Out of pocket, expenses were as high as dollars 3000/year for the 'poor health' group by year 3. Through analysis of the broad impact of PD, including non-neurological symptoms and economic ramifications, it is possible to better appreciate the impact of this chronic condition on overall quality of life. PMID- 11472880 TI - Positron emission tomography of dopamine pathways in familial Parkinsonian syndromes. AB - Positron emission tomography (PET) scan is considered to be the most useful tool with which to assess the integrity of nigrostriatal function in the living brain. Recently, different genetic defects have been associated with a variety of familial parkinsonian syndromes, the clinical phenotypes of which have varying degrees of similarities to idiopathic parkinsonism (IP), (sporadic Parkinson's disease). This review summarizes: (1) the PET scan findings (fluorodopa uptake and raclopride binding) in both familial parkinsonian syndromes and IP; and (2) the similarities and differences of the clinical and PET features between familial parkinsonian syndromes and IP. This analysis demonstrates that more similarities than differences exist in PET scan findings in the different familial parkinsonian syndromes with the exception of pallido-ponto-nigral degeneration (PPND), that is perhaps best considered as multisystem degeneration. As a result of this analysis, we believe that while different genetic defects may underlie different mechanisms of nigrostriatal degeneration, the final pattern of nigrostriatal dysfunction is essentially similar to that of IP. 'Parkinson's disease', therefore, may not represent a single disease entity, but rather the final manifestation of different pathogenetic mechanisms-mediated by genetic or environmental factors, or an interaction of genetic and environmental factors. PMID- 11472881 TI - Muscle weakness in Parkinson's disease: a follow-up study. AB - We compared isokinetic muscle strength between initial and subsequent measurements in 10 patients who could repeat the same testing later among 23 previously described patients with Parkinson's disease. Patients were divided into two groups according to changes in clinical condition between the times of the first and the subsequent measurements. For patients who had improved, both extension and flexion on the more affected side showed significantly greater torque at 15 rpm in the later than the earlier measurement. For patients who had worsened, both extension and flexion on the more affected side showed significantly less torque at 5 and 15 rpm in the later than the earlier measurement. Although isokinetic muscle strength is likely to depend on movement velocity in the early stages of Parkinson's disease, it may be influenced by bradykinesia, as the disease progresses. Speed-force correlation seen in these patients may give clues to the understanding of the pathophysiology of bradykinesia. PMID- 11472882 TI - Cortical Lewy bodies. PMID- 11472883 TI - A cross-over clinical and electromyographic assessment of treatment for parkinsonian tremor. AB - BACKGROUND: Whether dopaminergic and anticholinergic drugs exert influence on parkinsonian rest and postural tremor is a subject of debate. Different types of tremor may be influenced differently by the drugs. The aim of this study was to reevaluate the differential effects of levodopa and anticholinergic drugs on parkinsonian tremor in different limb positions and on different types of postural tremor. METHODS: Thirty-eight patients with parkinsonian resting tremor and postural tremor were included in this study. Patients were divided into two groups according to the electromyographic pattern of the postural tremor. We found fast synchronous postural tremor (>7 Hz) in 16 patients, and slow alternating postural tremor in 22 patients. The tremor was scored clinically in each limb position using the Webster Tremor Scale. Surface electromyographic recordings from the most involved limb in all positions were also performed. The patients were randomly assigned to levodopa (one 250/50-mg tablet), or to biperiden (one 3-mg tablet). Tremor was assessed by clinical and electromyographic examinations at base line 1h following ingestion of the drug. The subjective tremor improvement was also assessed. RESULTS: We found that levodopa had a good effect on the amplitude of the resting tremor, while the effect of biperiden was weaker. Both levodopa and biperiden has less effect on postural tremor. However, levodopa's effect was better than that of biperiden. Levodopa and biperiden had better effect on slow alternating postural tremor than on fast synchronous postural tremor. They had no effect on kinetic and intention tremors. CONCLUSIONS: Levodopa and anticholinergic drugs have differing effects on both resting and postural tremor Also, the different categories of postural tremor respond differently to treatment. The mechanisms underlying resting parkinsonian tremor may be different from those underlying postural, kinetic and intention tremor. Moreover, the mechanisms underlying different types of postural tremor may be different. PMID- 11472884 TI - Nonhuman primate models in skeletal research. PMID- 11472885 TI - Levormeloxifene prevents increased bone turnover and vertebral bone loss following ovariectomy in cynomolgus monkeys. AB - Levormeloxifene, a nonsteroidal selective estrogen receptor modulator (SERM), has been evaluated for its effects on bone in cynomolgus monkeys (Macaca fascicularis). Adult female monkeys were imported from Indonesia and randomized into six groups of 25-28 animals each (n = 158). Animals in one group were sham ovariectomized (sham) and received vehicle. Animals in the remaining five groups were ovariectomized and received either vehicle (ovx); 17beta-estradiol at 0.016 mg/kg (est); or levormeloxifene at 0.5 (L1), 1 (L2), or 5 (L3) mg/kg. Lumbar spine and whole body bone mass were measured by dual-energy X-ray absorptiometry (DXA) pretreatment and at 6 and 12 months following the initiation of treatment. Bone mass at the femoral neck was measured by peripheral quantitative computed tomography (pQCT) at 0 and 12 months. Serum markers of bone turnover, including bone-specific alkaline phosphatase (BSAP), osteocalcin (BGP), tartrate-resistant acid phosphatase (TRAP), and urinary collagen C-terminal extension peptides (CrossLaps), were measured at 0, 6, and 12 months. Ovariectomy resulted in an increase in these markers; the increase was prevented by estradiol or levormeloxifene. Estradiol or levormeloxifene inhibited loss of lumbar spine bone mineral density (BMD) following ovariectomy compared with untreated monkeys (ovx 5.0%; sham -0.4%; est +0.2%; L1 -3.6%, L2 -2.0%, L3 -2.5%). Estradiol, but not levormeloxifene, prevented loss of BMD at the femoral neck (ovx -7.4%; sham 3.1%; est -3.6%; L1 -8.0%, L2 -6.5%, L3 -7.8%), and whole body bone mineral content (BMC) (ovx -7.6%; sham -1.9%, est -2.9%; L1 -6.2%, L2 -6.1%, L3 -6.7%). Bone loss at each site was correlated with bone turnover as measured by serum and urine biomarkers. There was no dose effect of levormeloxifene. PMID- 11472886 TI - Evidence for targeted vesicular glutamate exocytosis in osteoblasts. AB - Regulated intercellular signaling is essential for the maintenance of bone mass. In recent work we described how osteoblasts and osteoclasts express functional receptors for the excitatory amino acid, glutamate, indicating that a signaling pathway analogous to synaptic neurotransmission exists in bone. Here, we show that osteoblasts also express the essential molecular framework for regulated glutamate exocytosis to occur as is present in presynaptic neurons. A combination of reverse transcription-polymerase chain reaction (RT-PCR) and northern and western blotting is used to show expression of the target membrane-SNARE (soluble NSF attachment protein receptor), proteins SNAP-25 and syntaxin 4 and the vesicular-SNARE protein VAMP (synaptobrevin), the minimum molecular requirements for core exocytotic complex formation. Immunofluorescent localizations reveal peripheral SNAP-25 expression on osteoblastic cells, particularly at intercellular contact sites, colocalizing with immunoreactive glutamate and the synaptic vesicle-specific protein, synapsin I. We also identify multiple accessory proteins associated with vesicle trafficking, including munc18, rSec8, DOC2, syntaxin 6, and synaptophysin, which have varied roles in regulated glutamate exocytosis. mRNA for the putative Ca(2+)-dependent regulators of vesicle recycling activity, synaptotagmin I (specialized for fast Ca(2+) dependent exocytosis as seen in synaptic neurotransmission), and the GTP-binding protein Rab3A are also identified by northern blot analysis. Finally, we demonstrate that osteoblastic cells actively release glutamate in a differentiation-dependent manner. These data provide compelling evidence that osteoblasts are able to direct glutamate release by regulated vesicular exocytosis, mimicking presynaptic glutamatergic neurons, showing that a process with striking similarity to synaptic neurotransmission occurs in bone. PMID- 11472887 TI - Role of endothelial nitric oxide synthase in estrogen-induced osteogenesis. AB - It is well recognized that high-dose estrogen induces a marked osteogenic response in long bones of female mice. In light of evidence which suggests that nitric oxide synthase (NOS) plays a role in regulation of osteoblast activity, we analyzed whether NOS is involved in mediating this response. Intact female mice were administered 17beta-estradiol (E(2)) either alone or in combination with N(G)-nitro-L-arginine methylester (L-NAME) or aminoguanidine (AG), over 24 days. The former inhibits both constitutive and inducible isoforms of NOS, whereas the latter is a selective inhibitor of inducible NOS. Bone mineral density (BMD) of the femur was subsequently measured by dual-energy X-ray absorptiometry (DXA), and histomorphometry performed at the proximal metaphysis on longitudinal tibial sections. As expected, E(2) given alone led to a marked accumulation of cancellous bone at the proximal tibial metaphysis, associated with a significant gain in femoral BMD, and an increase in cancellous mineralizing surfaces as assessed by histomorphometry. Neither L-NAME nor AG affected cancellous histomorphometric indices when given alone. However, when administered in combination with L-NAME, the magnitude of the skeletal response to E(2) was significantly reduced. The tendency for L-NAME to reduce estrogen-induced bone formation within the proximal tibial metaphysis was more marked distally compared with proximally. In contrast, AG showed no tendency to suppress the osteogenic response to E(2). Subsequently, we examined the effect of E(2) administration on expression within mouse femoral bone marrow of endothelial NOS (eNOS), which is the predominant constitutive isoform of NOS within bone. No change in eNOS mRNA levels was observed following E(2) administration, as assessed by reverse transcription-polymerase chain reaction (RT-PCR). Taken together, our results suggest that eNOS plays a role in mediating estrogen-induced bone formation in intact female mice, possibly as a consequence of posttranscriptional regulation of eNOS activity by estrogen. PMID- 11472888 TI - The effect of 17beta-estradiol on production of cytokines in cultures of peripheral blood. AB - Estrogen's action on bone may be mediated by cytokines produced by monocytes. We have reported a decreased ratio of interleukin-1beta (IL-1beta) to interleukin-1 receptor antagonist (IL-1ra) produced by whole blood cultures in vivo in women taking hormone replacement therapy (HRT). Also, one study has shown an effect of estradiol on tumor necrosis factor-alpha (TNF-alpha) secretion by separated monocytes in vitro. The aim of this study was to evaluate the effect of estrogen in vitro on the secretion of cytokines using whole blood cultures. Subjects consisted of 12 healthy postmenopausal women, ages 57-69 years, 4-20 years since menopause. Cytokines IL-1beta, interleukin-1alpha (IL-1alpha), IL-1ra, interleukin-6 (IL-6), TNF-alpha, and granulocyte macrophage-colony stimulating factor (GM-CSF) were measured in unstimulated and in stimulated (500 ng/mL lipopolysaccharide [LPS]) whole blood cultures treated with 17beta-estradiol (E(2)) at concentrations of 10(-12)--10(-6) mol/L. We found significant decreases in the spontaneous secretion of IL-6, TNF-alpha, IL-1ra, IL-1beta, and ratio of IL-1beta/IL-1ra compared with control, at physiological concentrations of E(2). The action of E(2) was blocked by the use of the antiestrogen ICI 182780 in coculture. A decrease in cytokine secretion was not observed when the inactive form of estrogen, 17alpha-estradiol, was used in place of 17beta-estradiol. GM CSF and IL-1alpha were not detectable in unstimulated cultures. Cytokine levels measured in stimulated cultures were not attenuated by treatment with E(2). We conclude that E(2) inhibits the spontaneous secretion of cytokines measured in whole blood cultures at physiological concentrations, and that the powerful stimulatory effect of LPS prevents any significant inhibition by E(2) in stimulated cultures. PMID- 11472889 TI - Changes in osteocalcin response to 1,25-dihydroxyvitamin D(3) stimulation and basal vitamin D receptor expression in human osteoblastic cells according to donor age and skeletal origin. AB - Age-related osteopenia is known to occur differently throughout the skeleton. In the present study, we examine the influence of donor age (<50 and >50 years), and bone structure (cortical vs. trabecular) on osteocalcin and vitamin D receptor (VDR) expression in primary cultures of human osteoblastic cells (hOB) cells. Cells were isolated from trabecular bone samples obtained from donors undergoing either knee (mainly trabecular) (n = 22; 4 <50 years, 18 >50 years) or hip (mainly cortical) (n = 16; 6 <50 years, 10 >50 years) arthroplasty. Pooling the results from knee and hip hOB cell cultures, we found that secreted osteocalcin was higher in hOB cells from the younger donors, compared with that in older donors, and peaked after stimulation with 10(-6)--10(-8) mol/L 1,25 dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. In cells from the latter donors, this secretion was maximal after 10(-6) mol/L 1,25(OH)(2)D(3) treatment. On the other hand, using reverse transcription followed by polymerase chain reaction, baseline osteocalcin mRNA was found to be lower in hOB cells from the older donors than in those from younger donors. After treatment with 10(-6)--10(-8) mol/L 1,25(OH)(2)D(3), osteocalcin mRNA increased over baseline in all groups of hOB cells studied. In age-matched cultures, both basal and 10(-6)--10(-8) mol/L 1,25(OH)(2)D(3)-stimulated osteocalcin mRNA showed similar values in hOB cells from both skeletal sites in younger donors. However, in the older donors, baseline as well as 10(-8) mol/L 1,25(OH)(2)D(3)-stimulated osteocalcin mRNA were higher in knee hOB cells than in hip hOB cells. Furthermore, baseline VDR mRNA expression was also higher in the former cells than in the latter cells in the older group. Considering the influence of donor age at each skeletal site of origin, we found lower baseline osteocalcin and VDR mRNA levels in hip hOB cells in the older group than in the younger group. Our findings indicate that the response of osteocalcin secretion and its mRNA to physiological doses of 1,25(OH)(2)D(3) decreases with aging and is associated with decreased VDR mRNA expression in hOB cells from mainly cortical bone. PMID- 11472890 TI - Comparison in localization between cystatin C and cathepsin K in osteoclasts and other cells in mouse tibia epiphysis by immunolight and immunoelectron microscopy. AB - We compared the distribution of a cysteine proteinase inhibitor, cystatin C, with that of cathepsin K in osteoclasts of the mouse tibia by immunolight and immunoelectron microscopy. Light microscopically, strong immunoreactivity for cystatin C was found extracellularly along the resorption lacuna and intracellularly in the organelles of osteoclasts. In serial sections, various patterns of cystatin C and cathepsin K localization were seen, specifically: (1) some resorption lacuna were positive for both cystatin C and cathepsin K; (2) others were positive for either cystatin C or cathepsin K, but not both; and (3) some lacuna were negative for both. In osteoclasts, the localization of cystatin C was similar to that of cathepsin K. Furthermore, cystatin C immunoreactivity was detected in preosteoclasts and osteoblasts, whereas cathepsin K was seen only in preosteoclasts. Electron microscopically, cystatin C immunoreactive products were found in the rough endoplasmic reticulum (ER), Golgi apparatus, vesicles, granules, and vacuoles of osteoclasts. These cystatin C-positive vesicles had fused or were in the process of fusion with the ampullar vacuoles (extracellular spaces) containing cystatin C-positive, fragmented, fibril-like structures. The extracellular cystatin C was deposited on and between the cytoplasmic processes of ruffled borders, and on and between type I collagen fibrils. In the basolateral region of osteoclasts, cystatin C-positive vesicles and granules also fused with vacuoles that contained cystatin C-positive or negative fibril-like structures. These results indicate that osteoclasts not only synthesize and secrete cathepsin K from the ruffled border into the bone resorption lacunae, but also a cysteine proteinase inhibitor, cystatin C. Therefore, it is suggested that cystatin C regulates the degradation of bone matrix by cathepsin K, both extracellularly and intracellularly. PMID- 11472891 TI - Characterization of human bone morphogenetic protein (BMP)-4 and -7 gene promoters: activation of BMP promoters by Gli, a sonic hedgehog mediator. AB - Among the bone morphogenetic protein (BMP) family, which plays a crucial role not only in bone formation but also in development, BMP-2, -4, and -7 participate predominantly in various aspects. To undertake complex tasks, their expression is strictly controlled. In this study we isolated and analyzed the 5'-flanking regions of the human BMP-4 and -7 genes to elucidate the mechanism of their temporally and spatially specific expression. As for BMP-4 expression, a reverse transcription-polymerase chain reaction (RT-PCR) assay with specially designed sets of primers demonstrated that osteoblastic SaOS-2 and Hos cells expressed two types of transcripts comprising one of the 5'-untranslated first exons, whereas MG63 cells displayed only the transcript with the BMP-4 proximal first exon. Likewise, RT-PCR revealed that Hos and MG63 cells expressed BMP-7. Subsequent 5' RACE confirmed an alternative usage of the BMP-4 first exons with clustered multiple transcription start sites in the distal exon and the sole start site in the proximal exon. The transcription start site of the BMP-7 gene was found to be far upstream (764 bp) of the initiation ATG codon. We constructed a series of deletion mutants of fusions between these BMP promoters and the luciferase gene and examined their activity by transient transfection into osteoblastic Hos and renal COS-7 cells. The degree of distal and proximal BMP-4 promoter activity was in accordance with the expression level of the corresponding transcripts. Both distal and proximal BMP-4 promoters possessed suppressor elements that are operative only in Hos cells. The positive and negative elements identified in the BMP-7 promoter were more remarkably effective in Hos cells. The activities of the respective BMP-4 promoters and BMP-7 promoter were all stimulated upon the cotransfection of a potential sonic hedgehog (SHH) mediator, Gli1 or Gli3 into COS-7 cells, providing direct evidence that the Gli proteins are capable of inducing the BMP expression. Our systems are helpful for assessment of the complicated interactions of molecules involved in the skeletogenesis and developmental processes. PMID- 11472892 TI - Calcium metabolism and bone mass in female rabbits during skeletal maturation: effects of dietary calcium intake. AB - This study documents growth and dual-energy X-ray absorptiometry (DXA)-determined peak bone mass profiles in the rabbit model, and tests the hypothesis that rabbits show patterns of bone accretion similar to humans and thus may serve as a viable model for human bone physiology. It is also shown that dietary Ca intake affects peak bone mass and the temporal pattern of its attainment. Groups of weaned animals were administered two nutritionally complete but calcium-altered diets (0.5% or 1.0% Ca). We evaluated growth, bone mass accretion, and Ca metabolism from 20 to 56 weeks of age in both the 1.0% Ca and 0.5% Ca groups of rabbits. For each monthly period, we monitored body mass, naso-tail length, food consumption, and fecal output. In addition, we collected blood and 24 h urine samples for biochemical analyses, and measured bone mass variables of the lumbar spine with DXA. The 1.0% Ca group had a lower apparent fractional absorption of Ca and higher urinary Ca excretion, but retained more Ca than the 0.5% Ca group during the growth phase. Furthermore, the 1.0% Ca group had lower parathyroid hormone (PTH) and bone biochemical marker concentrations throughout the study than the 0.5% Ca group. The lower levels of PTH and bone markers of resorption and formation, may have resulted in a reduction in skeletal remodeling, and this physiological mechanism may have contributed to the 10% increase in peak bone mineral density of the lumbar spine in the 1.0% Ca group of rabbits. PMID- 11472893 TI - Osteoporosis in the Norwegian moose. AB - In the last decade, an increased frequency of fractures among wild moose in the southernmost parts of Norway has been reported. This study focuses on bone mass, mechanical strength, and metal levels in the metacarpal bone of moose hunted in Aust-Agder (AA) County in southern Norway compared with control areas (CA) in more eastern and northern parts. Metacarpal bone with soft tissue from 106 AA and 37 CA animals were excised after hunting. Bone mineral density (BMD) was measured using dual-energy X-ray absorptiometry (DXA) at the proximal joint (trabecular bone tissue) and 100 mm distally (cortical bone tissue). Both accuracy and reproducibility of the method in these large bones were tested. Biomechanical strength was carried out using a three point bending test in bones from nine AA and eight CA animals of the same age. Metal concentrations in bone tissue were determined by atomic absorption spectrophotometry (GF-AAS) and atomic emission spectrometry (ICP-AAS). BMD of the proximal region was 1.58 +/- 0.39 g/cm(2) (mean +/- SD) for AA, 1.88 +/- 0.58 g/cm(2) (p < 0.01) for CA, 2.08 +/- 0.36 g/cm(2) for distal AA, and 2.16 +/- 0.48 (not significant) for distal CA. In analysis of covariance (ANCOVA) tests with age and region as factors, together with weight as cofactor, there were significant effects on BMD in both the proximal and distal areas for all factors. The bending strength of metacarpal bone was 9378 +/- 1814 N and 16,707 +/- 3065 N (p < 0.01) in AA and CA animals, respectively. There was no differences in element content except for the concentration of lead, where the mean value for AA animals was 1.5 +/- 0.8 mg/kg and CA 0.4 +/- 0.3 mg/kg (p < 0.01). In conclusion, the animals from the southern part of Norway had lower bone mass, decreased mechanical strength, and an increased bone concentration of lead. PMID- 11472894 TI - The influence of bone volume fraction and ash fraction on bone strength and modulus. AB - Although bone strength and modulus are known to be influenced by both volume fraction and mineral content (ash fraction), the relative influence of these two parameters remains unknown. Single-parameter power law functions are used widely to relate bone volume or ash fraction to bone strength and elastic modulus. In this study we evaluate the potential for predicting bone mechanical properties with two-parameter power law functions of bone volume fraction (BV/TV) and ash fraction (alpha) of the form y = a(BV/TV)(b) alpha(c) (where y is either ultimate strength or elastic modulus). We derived an expression for bone volume fraction as a function of apparent density and ash fraction to perform a new analysis of data presented by Keller in 1994. Exponents b and c for the prediction of bone strength were found to be 1.92 +/- 0.02 and 2.79 +/- 0.09 (mean +/- SE), respectively, with r(2) = 0.97. The value of b was found to be consistent with that found previously, whereas the value of c was lower than values previously reported. For the prediction of elastic modulus we found b and c to be 2.58 +/- 0.02 and 2.74 +/- 0.13, respectively, with r(2) = 0.97. The exponent related to ash fraction was typically larger than that associated with bone volume fraction, suggesting that a change in mineral content will, in general, generate a larger change in bone strength and stiffness than a similar change in bone volume fraction. These findings are important for interpreting the results of antiresorptive drug treatments that can cause changes in both ash and bone volume fraction. PMID- 11472895 TI - Quantitative ultrasound at the calcaneus in premenopausal women and their postmenopausal mothers. AB - The aim of the study was to establish a relationship between mothers' and daughters' bone status. Forty-eight postmenopausal women and their 48 premenopausal daughters were evaluated. The analysis was made for the whole group and for two subgroups: 27 healthy mothers and their 27 daughters; and 21 fractured mothers and their 21 daughters. The subgroups were matched for age and years since menopause (YSM), and height, weight, and body mass index (BMI) did not differ significantly. Bone status was evaluated by ultrasound measurement at the heel using the Achilles system (Lunar, Madison, WI), which measures speed of sound (SOS [m/sec]) and broadband ultrasound attenuation (BUA [dB/MHz]). The Achilles software also calculates a stiffness index (SI [%]). Ultrasound values for BUA, SOS, stiffness index, and Z score were significantly lower both in mothers with previous fractures and in their daughters, compared with respective values in mothers without fractures and their daughters. Future values in daughters were predicted using a stepwise, multiple regression analysis separately in the whole group and in the two subgroups. Future values were predicted in two models taking into consideration mothers' present SOS, BUA, and age or present SOS, BUA, and YSM. In both models, daughters' present SOS, BUA, age, height, and weight were taken into consideration. Predictive values were found to be high for daughters of women having had fractures (r = 0.72-0.87, p = 0.015-0.00007, SEE = 6.0-15.8) and lower for all daughters studied (r = 0.38 0.62, p = 0.03-0.0001, SEE = 8.8-21.5). In daughters of mothers without past fractures, prediction was not possible. Heritability of ultrasound values in daughters of women with past fractures ranged between 52% and 76%, whereas in the whole group the range was 14%-40%. In conclusion, the data indicate that, as a group, the daughters of women with osteoporotic fracture are likely to be at an increased risk for fractures because they have relatively low ultrasound values. Their future ultrasound values can be predicted on the basis of a single ultrasound evaluation with the condition that there is a history of maternal past fracture. PMID- 11472896 TI - Multicenter collaborative study to calibrate salmon calcitonin by bioassay and high-performance liquid chromatography: establishment of the third international standard. AB - Salmon calcitonin (sCT) is widely used therapeutically in the treatment of patients with postmenopausal osteoporosis, Paget's disease, and some forms of hypercalcemia. Preparations of synthetic calcitonin peptides of high purity and reproducibility are now routinely produced and physicochemical methods, particularly reverse-phase high-performance liquid chromatography (RP-HPLC), are replacing the in vivo biological assay for monitoring and calibration. Although the bioassay is no longer required for routine batch control in Europe, calcitonin bioassays are still required in some countries and in the development of new products. Stocks of the Second International Standard (IS) for salmon calcitonin are now depleted and, to replace it with a new calibrant, an international collaborative study was organized in which the aims were to: determine the activity of the candidate sCT by in vivo bioassay in terms of the second IS; assess the stability of the preparation after accelerated thermal degradation; estimate the purity of the ampouled candidate preparation; and determine the sCT content in gravimetric units by HPLC. The HPLC data in terms of ampoule content were in good agreement giving an estimate of 23.1 (coefficient of variation [CV] 3.8%) microg per ampoule. The HPLC chromatograms revealed a small, but detectable, degree of heterogeneity, which possibly occurred during the formulating or ampouling procedures, resulting in a reduction in monocomponent content (purity) from 96% to 92%. The biological activity of the ampoule contents in international units (IU) was calculated from the mass value and the internationally agreed-upon figure of 6000 IU/per mg for the specific activity of salmon calcitonin. This gave a value of 138 IU per ampoule, which was in good agreement with the biological assay estimate (140 IU per ampoule). The preparation of sCT was subsequently adopted as the Third International Standard by the World Health Organization with an assigned content of 138 IU per ampoule. PMID- 11472897 TI - Osteoporosis in men: a potential role for the sex hormone binding globulin. AB - The exact mechanism of bone loss remains unknown in primary male osteoporosis. It has been suggested that estrogen and sex hormone binding globulin (SHBG) play a role in regulating bone turnover and bone mass in healthy men > 65 years of age. In the present study, 80 men (mean age 49.7 years) with bone mineral density >2.5 SD below the young adult value and 40 age-matched controls were recruited to evaluate the relationships between sex hormone levels, bone biochemical markers levels, and bone mineral density. Fasting serum samples were assayed for total and free testosterone total estradiol, and SHBG. The free androgen index, was calculated as: [total testosterone/SHBG * 100]. Bone remodeling was evaluated by measurement of urinary levels of the C-telopeptide of type I collagen (CTx) and free deoxypyridinoline (D-Pyr), serum osteocalcin, and bone-specific alkaline phosphatase (bSAP). There was no significant difference between controls and osteoporotic men according to age, body mass index (BMI), total testosterone, and estradiol. In contrast, serum SHBG level was significantly higher (+42.2%), whereas free androgen index was lower (-24.8%) in patients with primary or secondary osteoporosis. Testosterone and estradiol levels did not correlate with any bone resorption or bone formation markers. In contrast, stepwise linear regression analysis showed that SHBG was significantly correlated with D-Pyr (r = 0.45, p < 0.05) and CTx (r = 0.34, p < 0.05) in primary osteoporosis. In secondary osteoporosis, SHBG was correlated with D-Pyr (r = 0.48, p < 0.05) and bSAP (r = 0.55, p < 0.01). After adjustment for age and BMI, hip bone mineral density (BMD) was not associated with testosterone or estradiol but only with serum SHBG (r = -0.33, p < 0.01) in primary osteoporosis. The same relationship was observed in men with secondary osteoporosis (r = -0.34, p < 0.01). Among osteoporotic patients, spinal radiography showed at least one vertebral crush fracture in 36 men and none in 44. Serum SHBG concentration was significantly associated with the presence of vertebral fracture: the odds ratio was 2.0 (95% confidence interval [CI] 1.2-3.5) for an increase of one standard deviation of SHBG. In conclusion, the present study showed that serum SHBG concentration is increased in middle-aged men with primary or secondary osteoporosis and is correlated with bone remodeling markers, hip bone mineral density, and vertebral fracture risk. PMID- 11472898 TI - Estrogen receptor gene polymorphism and bone mineral density in postmenopausal Chinese women. AB - PvuII and XbaI restriction fragment length polymorphisms (RFLPs) for the estrogen receptor gene (ERG) and its relation to bone mineral density (BMD) were examined in 454 postmenopausal Chinese women, aged 55-79 years. The RFLPs were represented as P or p (PvuII) and X or x (XbaI), with capital letters signifying the absence of and small letters the presence of restriction sites. There was no significant difference in BMD between the PP, Pp, and pp genotypes. However, women of the XX genotype had significantly higher BMD at the spine than women of the Xx or xx genotype. The magnitude of the difference in BMD was 80% of a standard deviation (SD) for BMD in elderly women and 40% of a SD in postmenopausal women. There was no statistically significant interaction between the PvuII genotype and the XbaI genotype in determining BMD. We conclude that postmenopausal Chinese women who were homozygous for the XX genotype had slightly higher BMD than the others. However, the difference in BMD was small and was unlikely to have any clinical significance. The ERG is not a major determinant of BMD in Chinese women in Hong Kong. PMID- 11472901 TI - Biotransformation of various substituted aromatic compounds to chiral dihydrodihydroxy derivatives. AB - The biotransformation of four different classes of aromatic compounds by the Escherichia coli strain DH5alpha(pTCB 144), which contained the chlorobenzene dioxygenase (CDO) from Pseudomonas sp. strain P51, was examined. CDO oxidized biphenyl as well as monochlorobiphenyls to the corresponding cis-2,3-dihydro-2,3 dihydroxy derivatives, whereby oxidation occurred on the unsubstituted ring. No higher substituted biphenyls were oxidized. The absolute configurations of several monosubstituted cis-benzene dihydrodiols formed by CDO were determined. All had an S configuration at the carbon atom in meta position to the substituent on the benzene nucleus. With one exception, the enantiomeric excess of several 1,4-disubstituted cis-benzene dihydrodiols formed by CDO was higher than that of the products formed by two toluene dioxygenases. Naphthalene was oxidized to enantiomerically pure (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene. All absolute configurations were identical to those of the products formed by toluene dioxygenases of Pseudomonas putida UV4 and P. putida F39/D. The formation rate of (+)-cis-(1R,2S)-dihydroxy-1,2-dihydronaphthalene was significantly higher (about 45 to 200%) than those of several monosubstituted cis-benzene dihydrodiols and more than four times higher than the formation rate of cis-benzene dihydrodiol. A new gas chromatographic method was developed to determine the enantiomeric excess of the oxidation products. PMID- 11472902 TI - Ecological aspects of ntcA gene expression and its use as an indicator of the nitrogen status of marine Synechococcus spp. AB - Nitrogen nutrition in cyanobacteria is regulated by NtcA, a transcriptional activator that is subject to negative control by ammonium. Using Synechococcus sp. strain WH7803 as a model organism, we show that ntcA expression was induced when cells were exposed to nitrogen stress but not when they were subjected to phosphorus or iron deprivation. Transcript levels accumulated in cells grown on a variety of inorganic and organic nitrogen sources, with the sole exception of ammonium. ntcA transcription was induced when ammonium levels dropped below 1 microM and reached maximal levels within 2 h. Furthermore, the addition of more than 1 microM ammonium led to a rapid decline in ntcA mRNA. The negative effect of ammonium was prevented by the addition of L-methionine-D,L-sulfoximine (MSX) and azaserine, inhibitors of ammonium assimilation. Thus, basal ntcA transcript levels are indicative of ammonium utilization. Conversely, the highest ntcA transcript levels were found in cells lacking a nitrogen source capable of supporting growth. Therefore, maximal ntcA expression would indicate nitrogen deprivation. This state of nitrogen deprivation was induced by a 1-h incubation with MSX. The rapid response of ntcA gene expression to the addition of ammonium and MSX was used to design a protocol for assessing relative ntcA transcript levels in field populations of cyanobacteria, from which their nitrogen status can be inferred. ntcA was basally expressed in Synechococcus at a nutrient enriched site at the northern tip of the Gulf of Aqaba, Red Sea. Therefore, these cyanobacteria were not nitrogen stressed, and their nitrogen requirements were met by regenerated nitrogen in the form of ammonium. PMID- 11472903 TI - Quantitative analysis of bacterial gene expression by using the gusA reporter gene system. AB - An Azospirillum brasilense Sp7 strain containing a plasmid-borne translational cytN-gusA fusion was grown in a continuous culture to quantitatively evaluate the influence of extracellular signals (such as O(2)) on expression of the cytNOQP operon. The dissolved oxygen concentration was shifted at regular time intervals before the steady state was reached. The measured beta-glucuronidase activity was used to monitor cytN gene expression. However, as the beta-glucuronidase activity in the experimental setup not only depended on altered transcription of the hybrid gene when the signal was varied but was also influenced by cellular accumulation, degradation, and dilution of the hybrid fusion protein, a mathematical method was developed to describe the intrinsic properties of the dynamic bioprocess. After identification and validation of the mathematical model, the apparent specific rate of expression of the fusion, which was independent of the experimental setup, could be deduced from the model and used to quantify gene expression regulated by extracellular environmental signals. In principle, this approach can be generalized to assess the effects of external signals on bacterial gene expression. PMID- 11472904 TI - Growth of chitinolytic dune soil beta-subclass Proteobacteria in response to invading fungal hyphae. AB - It has frequently been reported that chitinolytic soil bacteria, in particular biocontrol strains, can lyse living fungal hyphae, thereby releasing potential growth substrate. However, the conditions used in such assays (high bacterial density, rich media, fragmented hyphae) make it difficult to determine whether mycolytic activity is actually of importance for the growth and survival of chitinolytic bacteria in soils. An unidentified group of beta-subclass Proteobacteria (CbetaPs) was most dominant among the culturable nonfilamentous chitinolytic bacteria isolated from Dutch sand dune soils. Here we demonstrate that the CbetaPs grew at the expense of extending fungal mycelium of three dune soil fungi (Chaetomium globosum, Fusarium culmorum, and Mucor hiemalis) under nutrient-limiting, soil-like conditions. Aggregates of CbetaPs were also often found attached to fungal hyphae. The growth of a control group of dominant nonchitinolytic dune soil bacteria (beta- and gamma-subclass Proteobacteria) was not stimulated in the mycelial zone, indicating that growth-supporting materials were not independently released in appreciable amounts by the extending hyphae. Therefore, mycolytic activities of CbetaPs have apparently been involved in allowing them to grow after exposure to living hyphae. The chitinase inhibitor allosamidin did not, in the case of Mucor, or only partially, in the cases of Chaetomium and Fusarium, repress mycolytic growth of the CbetaPs, indicating that chitinase activity alone could not explain the extent of bacterial proliferation. Chitinolytic Stenotrophomonas-like and Cytophaga-like bacteria, isolated from the same dune soils, were only slightly stimulated by exposure to fungal hyphae. PMID- 11472905 TI - Carbon limitation induces sigma(S)-dependent gene expression in Pseudomonas fluorescens in soil. AB - Recent studies employing reporter gene technology indicate that the availabilities of the major nutrients nitrogen, phosphate, and iron to Pseudomonas are not severely limited in bulk soil. Indirect evidence has pointed to carbon limitation as a severe nutritional stress in this environment. We show that a plasmid (pGM115)-borne transcriptional fusion between the sigma(S) dependent Escherichia coli promoter P(fic) and lacZ functions as a reliable reporter for carbon availability in Pseudomonas fluorescens. When P. fluorescens strain DF57(pGM115) was introduced into bulk soil, carbon-limiting conditions were indicated by citrate-repressible induction of beta-galactosidase activity. To address carbon availability at the single-cell level, we developed an immunofluorescence double-staining procedure for individual DF57 cells expressing beta-galactosidase from P(fic). Changes in cell size and expression of beta galactosidase were analyzed by flow cytometry. Cells extracted from soil microcosms reduced their size less than carbon-starved cells in pure culture and showed an increased tendency to aggregate. The single-cell analysis revealed that for cells residing in soil, the expression of beta-galactosidase became heterogeneous and only a DF57 subpopulation appeared to be carbon limited. In soil amended with barley straw, limited nitrogen availability has been determined by use of the bioluminescent reporter strain P. fluorescens DF57-N3. We used strain DF57-N3(pGM115) as a double reporter for carbon and nitrogen limitation that allowed us to study the dynamics of carbon and nitrogen availabilities in more detail. In straw-amended soil beta-galactosidase activity remained low, while nitrogen limitation-dependent bioluminescence appeared after a few days. Hence, nitrogen became limited under conditions where carbon resources were not completely exhausted. PMID- 11472906 TI - Effect of genetically modified Pseudomonas putida WCS358r on the fungal rhizosphere microflora of field-grown wheat. AB - We released genetically modified Pseudomonas putida WCS358r into the rhizospheres of wheat plants. The two genetically modified derivatives, genetically modified microorganism (GMM) 2 and GMM 8, carried the phz biosynthetic gene locus of strain P. fluorescens 2-79 and constitutively produced the antifungal compound phenazine-1-carboxylic acid (PCA). In the springs of 1997 and 1998 we sowed wheat seeds treated with either GMM 2, GMM 8, or WCS358r (approximately 10(7) CFU per seed), and measured the numbers, composition, and activities of the rhizosphere microbial populations. During both growing seasons, all three bacterial strains decreased from 10(7) CFU per g of rhizosphere sample to below the limit of detection (10(2) CFU per g) 1 month after harvest of the wheat plants. The phz genes were stably maintained, and PCA was detected in rhizosphere extracts of GMM treated plants. In 1997, but not in 1998, fungal numbers in the rhizosphere, quantified on 2% malt extract agar (total filamentous fungi) and on Komada's medium (mainly Fusarium spp.), were transiently suppressed in GMM 8-treated plants. We also analyzed the effects of the GMMs on the rhizosphere fungi by using amplified ribosomal DNA restriction analysis. Introduction of any of the three bacterial strains transiently changed the composition of the rhizosphere fungal microflora. However, in both 1997 and 1998, GMM-induced effects were distinct from those of WCS358r and lasted for 40 days in 1997 and for 89 days after sowing in 1998, whereas effects induced by WCS358r were detectable for 12 (1997) or 40 (1998) days. None of the strains affected the metabolic activity of the soil microbial population (substrate-induced respiration), soil nitrification potential, cellulose decomposition, plant height, or plant yield. The results indicate that application of GMMs engineered to have improved antifungal activity can exert nontarget effects on the natural fungal microflora. PMID- 11472907 TI - Assessment of the genetic diversity among strains of Xanthomonas cynarae by randomly amplified polymorphic DNA analysis and development of specific characterized amplified regions for the rapid identification of X. cynarae. AB - The randomly amplified polymorphic DNA (RAPD) method was used to investigate the genetic diversity in Xanthomonas cynarae, which causes bacterial bract spot disease of artichoke. This RAPD analysis was also intended to identify molecular markers characteristic of this species, in order to develop PCR-based markers which can be used to detect this pathogenic bacterium in artichoke fields. Among the 340 RAPD primers tested, 40 were selected on their ability to produce reproducible and reliable fingerprints in our genetic background. These 40 primers produced almost similar patterns for the 37 X. cynarae strains studied, different from the fingerprints obtained for other Xanthomonas species and other xanthomonad-like bacteria isolated from artichoke leaves. Therefore, X. cynarae strains form a homogeneous genetic group. However, a little DNA polymorphism within this species was observed and the collection of X. cynarae isolates was divided into two groups (one containing three strains, the second one including all other strains). Out of seven RAPD markers characteristic of X. cynarae that were cloned, four did not hybridize to the genomic DNA of strains belonging to other Xanthomonas species. These four RAPD markers were converted into PCR markers (specific characterized amplified regions [SCARs]); they were sequenced, and a PCR primer pair was designed for each of them. Three derived SCARs are good candidates to develop PCR-based tests to detect X. cynarae in artichoke fields. PMID- 11472908 TI - Relationship between monokaryotic growth rate and mating type in the edible basidiomycete Pleurotus ostreatus. AB - The edible fungus Pleurotus ostreatus (oyster mushroom) is an industrially produced heterothallic homobasidiomycete whose mating is controlled by a bifactorial tetrapolar genetic system. Two mating loci (matA and matB) control different steps of hyphal fusion, nuclear migration, and nuclear sorting during the onset and progress of the dikaryotic growth. Previous studies have shown that the segregation of the alleles present at the matB locus differs from that expected for a single locus because (i) new nonparental B alleles appeared in the progeny and (ii) there was a distortion in the segregation of the genomic regions close to this mating locus. In this study, we pursued these observations by using a genetic approach based on the identification of molecular markers linked to the matB locus that allowed us to dissect it into two genetically linked subunits (matBalpha and matBbeta) and to correlate the presence of specific matBalpha and matA alleles with differences in monokaryotic growth rate. The availability of these molecular markers and the mating type dependence of growth rate in monokaryons can be helpful for marker-assisted selection of fast-growing monokaryons to be used in the construction of dikaryons able to colonize the substrate faster than the competitors responsible for reductions in the industrial yield of this fungus. PMID- 11472909 TI - Effects of nondigestible oligosaccharides on Salmonella enterica serovar Typhimurium and nonpathogenic Escherichia coli in the pig small intestine in vitro. AB - An in vitro intestinal tissue model was developed for the investigation of bacterial association in the pig small intestine under different dietary regimes. In preliminary experiments, jejunal and ileal tissue was taken from Danish Landrace pigs fed standard diet and inoculated with either Salmonella or nonpathogenic Escherichia coli strains. Higher numbers of salmonellae associated with the ileal tissues, but the numbers did not reach significance. Hence, jejunal sections were inoculated with nonpathogenic E. coli and ileal sections were inoculated with salmonellae in the presence of mannose or commercial nondigestible oligosaccharides (NDO) at 2.5%. There was a significant decrease in E. coli associated with the jejunum in the presence of mannose (P < 0.05). Furthermore, in pigs fed a diet supplemented with commercial NDO at 4% there was a significant reduction in the numbers of E. coli in jejunal organ cultures of pigs fed the FOS diet (P < 0.05). There was a reduction, though not a significant one, in the association of Salmonella sp. to the ileal sections of pigs fed the commercial FOS diet. The feeding of commercial GOS or its addition to organ cultures did not affect E. coli or Salmonella numbers. PMID- 11472910 TI - Altered protein expression of Streptococcus oralis cultured at low pH revealed by two-dimensional gel electrophoresis. AB - Streptococcus oralis is the predominant aciduric nonmutans streptococcus isolated from the human dentition, but the role of this organism in the initiation and progression of dental caries has yet to be established. To identify proteins that are differentially expressed by S. oralis growing under conditions of low pH, soluble cellular proteins extracted from bacteria grown in batch culture at pH 5.2 or 7.0 were analyzed by two-dimensional (2-D) gel electrophoresis. Thirty nine proteins had altered expression at low pH; these were excised, digested with trypsin using an in-gel protocol, and further analyzed by peptide mass fingerprinting using matrix-assisted laser desorption ionization mass spectrometry. The resulting fingerprints were compared with the genomic database for Streptococcus pneumoniae, an organism that is phylogenetically closely related to S. oralis, and putative functions for the majority of these proteins were determined on the basis of functional homology. Twenty-eight proteins were up-regulated following growth at pH 5.2; these included enzymes of the glycolytic pathway (glyceraldehyde-3-phosphate dehydrogenase and lactate dehydrogenase), the polypeptide chains comprising ATP synthase, and proteins that are considered to play a role in the general stress response of bacteria, including the 60-kDa chaperone, Hsp33, and superoxide dismutase, and three distinct ABC transporters. These data identify, for the first time, gene products that may be important in the survival and proliferation of nonmutans aciduric S. oralis under conditions of low pH that are likely to be encountered by this organism in vivo. PMID- 11472911 TI - Effects of iron limitation on the degradation of toluene by Pseudomonas strains carrying the tol (pWWO) plasmid. AB - Most aerobic biodegradation pathways for hydrocarbons involve iron-containing oxygenases. In iron-limited environments, such as the rhizosphere, this may influence the rate of degradation of hydrocarbon pollutants. We investigated the effects of iron limitation on the degradation of toluene by Pseudomonas putida mt2 and the transconjugant rhizosphere bacterium P. putida WCS358(pWWO), both of which contain the pWWO (TOL) plasmid that harbors the genes for toluene degradation. The results of continuous-culture experiments showed that the activity of the upper-pathway toluene monooxygenase decreased but that the activity of benzyl alcohol dehydrogenase was not affected under iron-limited conditions. In contrast, the activities of three meta-pathway (lower-pathway) enzymes were all found to be reduced when iron concentrations were decreased. Additional experiments in which citrate was used as a growth substrate and the pathways were induced with the gratuitous inducer o-xylene showed that expression of the TOL genes increased the iron requirement in both strains. Growth yields were reduced and substrate affinities decreased under iron-limited conditions, suggesting that iron availability can be an important parameter in the oxidative breakdown of hydrocarbons. PMID- 11472912 TI - Decrease in cell surface galactose residues of Schizosaccharomyces pombe enhances its coflocculation with Pediococcus damnosus. AB - Pediococcus damnosus can coflocculate with Saccharomyces cerevisiae and cause beer acidification that may or may not be desired. Similar coflocculations occur with other yeasts except for Schizosaccharomyces pombe which has galactose-rich cell walls. We compared coflocculation rates of S. pombe wild-type species TP4 1D, having a mannose-to-galactose ratio (Man:Gal) of 5 to 6 in the cell wall, with its glycosylation mutants gms1-1 (Man:Gal = 5:1) and gms1Delta (Man:Gal = 1:0). These mutants coflocculated at a much higher level (30 to 45%) than that of the wild type (5%). Coflocculation of the mutants was inhibited by exogenous mannose but not by galactose. The S. cerevisiae mnn2 mutant, with a mannan content similar to that of gms1Delta, also showed high coflocculation (35%) and was sensitive to mannose inhibition. Coflocculation of P. damnosus and gms1Delta (or mnn2) also could be inhibited by gms1Delta mannan (with unbranched alpha-1,6 linked mannose residues), concanavalin A (mannose and glucose specific), or NPA lectin (specific for alpha-1,6-linked mannosyl units). Protease treatment of the bacterial cells completely abolished coflocculation. From these results we conclude that mannose residues on the cell surface of S. pombe serve as receptors for a P. damnosus lectin but that these receptors are shielded by galactose residues in wild-type strains. Such interactions are important in the production of Belgian acid types of beers in which mixed cultures are used to improve flavor. PMID- 11472913 TI - Impact of 2,4-diacetylphloroglucinol-producing biocontrol strain Pseudomonas fluorescens F113 on intraspecific diversity of resident culturable fluorescent pseudomonads associated with the roots of field-grown sugar beet seedlings. AB - The impact of the 2,4-diacetylphloroglucinol-producing biocontrol agent Pseudomonas fluorescens F113Rif on the diversity of the resident community of culturable fluorescent pseudomonads associated with the roots of field-grown sugar beet seedlings was evaluated. At 19 days after sowing, the seed inoculant F113Rif had replaced some of the resident culturable fluorescent pseudomonads at the rhizoplane but had no effect on the number of these bacteria in the rhizosphere. A total of 498 isolates of resident fluorescent pseudomonads were obtained and characterized by molecular means at the level of broad phylogenetic groups (by amplified ribosomal DNA restriction analysis) and at the strain level (with random amplified polymorphic DNA markers) as well as phenotypically (55 physiological tests). The introduced pseudomonad induced a major shift in the composition of the resident culturable fluorescent Pseudomonas community, as the percentage of rhizoplane isolates capable of growing on three carbon substrates (erythritol, adonitol, and L-tryptophan) not assimilated by the inoculant was increased from less than 10% to more than 40%. However, the pseudomonads selected did not display enhanced resistance to 2,4-diacetylphloroglucinol. The shift in the resident populations, which was spatially limited to the surface of the root (i.e., the rhizoplane), took place without affecting the relative proportions of phylogenetic groups or the high level of strain diversity of the resident culturable fluorescent Pseudomonas community. These results suggest that the root associated Pseudomonas community of sugar beet seedlings is resilient to the perturbation that may be caused by a taxonomically related inoculant. PMID- 11472914 TI - Influence of effluent irrigation on community composition and function of ammonia oxidizing bacteria in soil. AB - The effect of effluent irrigation on community composition and function of ammonia-oxidizing bacteria (AOB) in soil was evaluated, using techniques of molecular biology and analytical soil chemistry. Analyses were conducted on soil sampled from lysimeters and from a grapefruit orchard which had been irrigated with wastewater effluent or fertilizer-amended water (FAW). Specifically, comparisons of AOB community composition were conducted using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified fragments of the gene encoding the alpha-subunit of the ammonia monooxygenase gene (amoA) recovered from soil samples and subsequent sequencing of relevant bands. A significant and consistent shift in the population composition of AOB was detected in soil irrigated with effluent. This shift was absent in soils irrigated with FAW, despite the fact that the ammonium concentration in the FAW was similar. At the end of the irrigation period, Nitrosospira-like populations were dominant in soils irrigated with FAW, while Nitrosomonas-like populations were dominant in effluent-irrigated soils. Furthermore, DGGE analysis of the amoA gene proved to be a powerful tool in evaluating the soil AOB community population and population shifts therein. PMID- 11472915 TI - Location effects of a reporter gene on expression levels and on native protein synthesis in Lactococcus lactis and Saccharomyces cerevisiae. AB - The engineering of industrially important genetically modified organisms by the integration of heterologous genes into the chromosome is often the method of choice for several reasons concerned with long-term stability, homogeneous population distribution, and the enabling of selection without the addition of antibiotics. However, integration may disrupt endogenous gene expression, giving rise to increased levels of toxic metabolic byproducts or activating otherwise silent genes. The position of integration of a foreign gene in the chromosome can also influence its expression levels, and this effect will be of relevance in terms of optimizing protein production parameters. In this study, we determine how the random integration of a foreign reporter gene might affect expression levels and assess the use of proteome analysis to investigate possible effects on synthesis of endogenous proteins in two important food-relevant microorganisms, Saccharomyces cerevisiae and Lactococcus lactis. Eleven L. lactis integrants carrying the gusA gene were analyzed, and expression levels were found to vary by a factor of threefold in contrast to expression levels of lacZ in 18 S. cerevisiae integrants, which showed a 14-fold variation. Of relevance to industry is whether any changes in expression levels might occur as a consequence of storage of the modified strains. Here it is also shown that the above differences in expression levels were not significantly affected by storage of frozen cultures over a period of several months. Analysis of the protein composition of the yeast and lactococcal integrant strains by separation on one-dimensional (1D) and 2D gels showed no significant variations in position beyond those observed in control samples. PMID- 11472916 TI - Laboratory-scale evidence for lightning-mediated gene transfer in soil. AB - Electrical fields and current can permeabilize bacterial membranes, allowing for the penetration of naked DNA. Given that the environment is subjected to regular thunderstorms and lightning discharges that induce enormous electrical perturbations, the possibility of natural electrotransformation of bacteria was investigated. We demonstrated with soil microcosm experiments that the transformation of added bacteria could be increased locally via lightning mediated current injection. The incorporation of three genes coding for antibiotic resistance (plasmid pBR328) into the Escherichia coli strain DH10B recipient previously added to soil was observed only after the soil had been subjected to laboratory-scale lightning. Laboratory-scale lightning had an electrical field gradient (700 versus 600 kV m(-1)) and current density (2.5 versus 12.6 kA m(-2)) similar to those of full-scale lightning. Controls handled identically except for not being subjected to lightning produced no detectable antibiotic-resistant clones. In addition, simulated storm cloud electrical fields (in the absence of current) did not produce detectable clones (transformation detection limit, 10(-9)). Natural electrotransformation might be a mechanism involved in bacterial evolution. PMID- 11472917 TI - Three glycoproteins with antimutagenic activity identified in Lactobacillus plantarum KLAB21. AB - Antimutagenic substances were purified from a culture supernatant of Lactobacillus plantarum KLAB21 cells isolated from kimchi, a Korean traditional fermented vegetable, and their characteristics were investigated. The antimutagenic substances were separated into two fractions by DEAE-cellulose ion exchange column chromatography, which were designated the R1 and R2 fractions. The R1 fraction was then divided into two fractions again by Sephadex G200 gel filtration chromatography, and the fractions were designated R1-1 and R1-2. All three fractions were further purified using a Sepharose CL-6B gel filtration column. All the purified fractions were successfully stained with fuchsin as well as Coomassie brilliant blue, suggesting that they are glycoproteins. The purified fractions were confirmed to possess antimutagenic activity against N-methyl-N' nitro-N-nitrosoguanidine on Salmonella enterica serovar Typhimurium TA100 cells. Their molecular masses were determined to be 16 (R1-1), 11 (R1-2), and 14 (R2) kDa on the Sepharose CL-6B column. Total sugar contents were 8.4% (R1-1), 7.3% (R1-2), and 9.4% (R2). The amino acid compositions of the fractions were different from each other; the major amino acids were glutamic acid (21.5%) and phenylalanine (17.1%) in the R1-1 fraction and glycine (41.3%) in the R1-2 fraction, but valine (31%) and phenylalanine (22.6%) were the major amino acids in the R2 fraction. PMID- 11472918 TI - Differentiation of Lactobacillus plantarum, L. pentosus, and L. paraplantarum by recA gene sequence analysis and multiplex PCR assay with recA gene-derived primers. AB - In this study, we succeeded in differentiating Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus paraplantarum by means of recA gene sequence comparison. Short homologous regions of about 360 bp were amplified by PCR with degenerate consensus primers, sequenced, and analyzed, and 322 bp were considered for the inference of phylogenetic trees. Phylograms, obtained by parsimony, maximum likelihood, and analysis of data matrices with the neighbor joining model, were coherent and clearly separated the three species. The validity of the recA gene and RecA protein as phylogenetic markers is discussed. Based on the same sequences, species-specific primers were designed, and a multiplex PCR protocol for the simultaneous distinction of these bacteria was optimized. The sizes of the amplicons were 318 bp for L. plantarum, 218 bp for L. pentosus, and 107 bp for L. paraplantarum. This strategy permitted the unambiguous identification of strains belonging to L. plantarum, L. pentosus, and L. paraplantarum in a single reaction, indicating its applicability to the speciation of isolates of the L. plantarum group. PMID- 11472919 TI - Isolation and characterization of a gene specific to lager brewing yeast that encodes a branched-chain amino acid permease. AB - We found two types of branched-chain amino acid permease gene (BAP2) in the lager brewing yeast Saccharomyces pastorianus BH-225 and cloned one type of BAP2 gene (Lg-BAP2), which is identical to that of Saccharomyces bayanus (by-BAP2-1). The other BAP2 gene of the lager brewing yeast (cer-BAP2) is very similar to the Saccharomyces cerevisiae BAP2 gene. This result substantiates the notion that lager brewing yeast is a hybrid of S. cerevisiae and S. bayanus. The amino acid sequence homology between S. cerevisiae Bap2p and Lg-Bap2p was 88%. The transcription of Lg-BAP2 was not induced by the addition of leucine to the growth medium, while that of cer-BAP2 was induced. The transcription of Lg-BAP2 was repressed by the presence of ethanol and weak organic acid, while that of cer BAP2 was not affected by these compounds. Furthermore, Northern analysis during beer fermentation revealed that the transcription of Lg-BAP2 was repressed at the beginning of the fermentation, while cer-BAP2 was highly expressed throughout the fermentation. These results suggest that the transcription of Lg-BAP2 is regulated differently from that of cer-BAP2 in lager brewing yeasts. PMID- 11472920 TI - Brown pigments produced by Yarrowia lipolytica result from extracellular accumulation of homogentisic acid. AB - Yarrowia lipolytica produces brown extracellular pigments that correlate with tyrosine catabolism. During tyrosine depletion, the yeast accumulated homogentisic acid, p-hydroxyphenylethanol, and p-hydroxyphenylacetic acid in the medium. Homogentisic acid accumulated under all aeration conditions tested, but its concentration decreased as aeration decreased. With moderate aeration, equimolar concentrations of alcohol and p-hydroxyphenylacetic acid (1:1) were detected, but with lower aeration the alcohol concentration was twice that of the acid (2:1). p-Hydroxyphenylethanol and p-hydroxyphenylacetic acid may result from the spontaneous disproportionation of the corresponding aldehyde, p hydroxyphenylacetaldehyde. The catabolic pathway of tyrosine in Y. lipolytica involves the formation of p-hydroxyphenylacetaldehyde, which is oxidized to p hydroxyphenylacetic acid and then further oxidized to homogentisic acid. Brown pigments are produced when homogentisic acid accumulates in the medium. This acid can spontaneously oxidize and polymerize, leading to the formation of pyomelanins. Mn(2+) accelerated and intensified the oxidative polymerization of homogentisic acid, and lactic acid enhanced the stimulating role of Mn(2+). Alkaline conditions also accelerated pigment formation. The proposed tyrosine catabolism pathway appears to be unique for yeast, and this is the first report of a yeast producing pigments involving homogentisic acid. PMID- 11472921 TI - Adhesion of bifidobacteria to granular starch and its implications in probiotic technologies. AB - Adhesion of 19 Bifidobacterium strains to native maize, potato, oat, and barley starch granules was examined to investigate links between adhesion and substrate utilization and to determine if adhesion to starch could be exploited in probiotic food technologies. Starch adhesion was not characteristic of all the bifidobacteria tested. Adherent bacteria bound similarly to the different types of starch, and the binding capacity of the starch (number of bacteria per gram) correlated to the surface area of the granules. Highly adherent strains were able to hydrolyze the granular starches, but not all amylolytic strains were adherent, indicating that starch adhesion is not a prerequisite for efficient substrate utilization for all bifidobacteria. Adhesion was mediated by a cell surface protein(s). For the model organisms tested (Bifidobacterium adolescentis VTT E 001561 and Bifidobacterium pseudolongum ATCC 25526), adhesion appeared to be specific for alpha-1,4-linked glucose sugars, since adhesion was inhibited by maltose, maltodextrin, amylose, and soluble starch but not by trehalose, cellobiose, or lactose. In an in vitro gastric model, adhesion was inhibited both by the action of protease and at pH values of < or =3. Adhesion was not affected by bile, but the binding capacity of the starch was reduced by exposure to pancreatin. It may be possible to exploit adhesion of probiotic bifidobacteria to starch granules in microencapsulation technology and for synbiotic food applications. PMID- 11472922 TI - Bile salt hydrolase activity and resistance to toxicity of conjugated bile salts are unrelated properties in lactobacilli. AB - Bacteria of numerous species isolated from the human gastrointestinal tract express bile salt hydrolase (BSH) activity. How this activity contributes to functions of the microorganisms in the gastrointestinal tract is not known. We tested the hypothesis that a BSH protects the cells that produce it from the toxicity of conjugated bile salts. Forty-nine strains of numerous Lactobacillus spp. were assayed to determine their capacities to express BSH activities (taurodeoxycholic acid [TDCA] hydrolase and taurocholic acid [TCA] hydrolase activities) and their capacities to resist the toxicity of a conjugated bile acid (TDCA). Thirty of these strains had been isolated from the human intestine, 15 had been recovered from dairy products, and 4 had originated from other sources. Twenty-six of the strains expressed both TDCA hydrolase and TCA hydrolase activities. One strain that expressed TDCA hydrolase activity did not express TCA hydrolase activity. Conversely, in one strain for which the assay for TDCA hydrolase activity gave a negative result there was evidence of TCA hydrolase activity. Twenty-five of the strains were found to resist the toxicity of TDCA. Fourteen of these strains were of human origin, nine were from dairy products, and two were from other sources. Of the 26 strains expressing both TDCA hydrolase and TCA hydrolase activities, 15 were resistant to TDCA toxicity, 6 were susceptible, and 5 gave inconclusive results. Of the 17 strains that gave negative results for either of the enzymes, 7 were resistant to the toxicity, 9 were susceptible, and 1 gave inconclusive results. These findings do not support the hypothesis tested. They suggest, however, that BSH activity is important at some level for lactobacillus colonization of the human intestine. PMID- 11472923 TI - Presence of nitrate-accumulating sulfur bacteria and their influence on nitrogen cycling in a shallow coastal marine sediment. AB - Nitrate flux between sediment and water, nitrate concentration profile at the sediment-water interface, and in situ sediment denitrification activity were measured seasonally at the innermost part of Tokyo Bay, Japan. For the determination of sediment nitrate concentration, undisturbed sediment cores were sectioned into 5-mm depth intervals and each segment was stored frozen at -30 degrees C. The nitrate concentration was determined for the supernatants after centrifuging the frozen and thawed sediments. Nitrate in the uppermost sediment showed a remarkable seasonal change, and its seasonal maximum of up to 400 microM was found in October. The directions of the diffusive nitrate fluxes predicted from the interfacial concentration gradients were out of the sediment throughout the year. In contrast, the directions of the total nitrate fluxes measured by the whole-core incubation were into the sediment at all seasons. This contradiction between directions indicates that a large part of the nitrate pool extracted from the frozen surface sediments is not a pore water constituent, and preliminary examinations demonstrated that the nitrate was contained in the intracellular vacuoles of filamentous sulfur bacteria dwelling on or in the surface sediment. Based on the comparison between in situ sediment denitrification activity and total nitrate flux, it is suggested that intracellular nitrate cannot be directly utilized by sediment denitrification, and the probable fate of the intracellular nitrate is hypothesized to be dissimilatory reduction to ammonium. The presence of nitrate-accumulating sulfur bacteria therefore may lower nature's self purification capacity (denitrification) and exacerbate eutrophication in shallow coastal marine environments. PMID- 11472924 TI - Characterization of the 13-kilobase ermF region of the Bacteroides conjugative transposon CTnDOT. AB - The conjugative transposon CTnDOT is virtually identical over most of its length to another conjugative transposon, CTnERL, except that CTnDOT carries an ermF gene that is not found on CTnERL. In this report, we show that the region containing ermF appears to consist of a 13-kb chimera composed of at least one class I composite transposon and a mobilizable transposon (MTn). Although the ermF region contains genes also carried on Bacteroides transposons Tn4351 and Tn4551, it does not contain the IS4351 element which is found on these transposons. In CTnDOT, insertion of the ermF region occurred near a stem-loop structure at the end of orf2, an open reading frame located immediately downstream of the integrase (int) gene of CTnDOT, and in a region known to be important for excision of CTnERL and CTnDOT. The chimera that comprises the ermF region can apparently no longer excise and circularize, but it contains a functional mobilization region related to that described for the Bacteroides MTn Tn4399. Analysis of 19 independent Bacteroides isolates showed that the ermF region is located in the same position in all of the strains analyzed and that the compositions of the ermF region are almost identical in these strains. Therefore, it appears that CTnDOT-like elements present in community and clinical isolates of Bacteroides were derived from a common ancestor and proliferated in the diverse Bacteroides population. PMID- 11472925 TI - Multiphasic kinetics of transformation of 1,2,4-trichlorobenzene at nano- and micromolar concentrations by Burkholderia sp. strain PS14. AB - The transformation of 1,2,4-trichlorobenzene (1,2,4-TCB) at initial concentrations in nano- and micromolar ranges was studied in batch experiments with Burkholderia sp. strain PS14. 1,2,4-TCB was metabolized from nano- and micromolar concentrations to below its detection limit of 0.5 nM. At low initial 1,2,4-TCB concentrations, a first-order relationship between specific transformation rate and substrate concentration was observed with a specific affinity (a(0)(A)) of 0.32 liter. mg (dry weight)(-1). h(-1) followed by a second one at higher concentrations with an a(o)(A) of 0.77 liter. mg (dry weight)(-1). h(-1). This transition from the first-order kinetics at low initial 1,2,4-TCB concentrations to the second first-order kinetics at higher 1,2,4-TCB concentrations was shifted towards higher initial 1,2,4-TCB concentrations with increasing cell mass. At high initial concentrations of 1,2,4-TCB, a maximal transformation rate of approximately 37 nmol. min(-1). mg (dry weight)(-1) was measured, irrespective of the cell concentration. PMID- 11472926 TI - Rapid evolution of a sexual reproduction gene in centric diatoms of the genus Thalassiosira. AB - Sexual reproduction is commonly assumed to occur in the vast majority of diatoms due to the intimate association of this process with cell size control. Surprisingly, however, little is known about the impact of sexual events on diatom population dynamics. The Sig1 gene is strongly upregulated during sexual reproduction in the centric diatom Thalassiosira weissflogii and has been hypothesized to encode a protein involved in gamete recognition. In the present study, degenerate PCR primers were designed and used to amplify a portion of Sig1 from three closely related species in the cosmopolitan genus Thalassiosira, Thalassiosira oceanica, Thalassiosira guillardii, and Thalassiosira pseudonana. Identification of Sig1 in these three additional species facilitated development of this gene as a molecular marker for diatom sexual events. Examination of the new sequences indicated that multiple copies of Sig1 are probably present in the genome. Moreover, compared to the housekeeping gene beta-tubulin, the Sig1 genes of isolates of T. weissflogii collected from different regions of the Atlantic and Pacific oceans displayed high levels of divergence. The Sig1 genes of the four closely related Thalassiosira species also displayed high levels of sequence divergence compared to the levels observed with a second gene, Fcp, probably explaining why Sig1 could not be amplified from more distantly related species. The high levels of sequence divergence both within and between species suggest that Sig1 is rapidly evolving in a manner reminiscent of the manner observed in other genes that encode gamete recognition proteins. A simple model is presented for Sig1 evolution and the implications of such a rapidly evolving sexual reproduction gene for diatom speciation and population dynamics. PMID- 11472927 TI - The P450-4 gene of Gibberella fujikuroi encodes ent-kaurene oxidase in the gibberellin biosynthesis pathway. AB - At least five genes of the gibberellin (GA) biosynthesis pathway are clustered on chromosome 4 of Gibberella fujikuroi; these genes encode the bifunctional ent copalyl diphosphate synthase/ent-kaurene synthase, a GA-specific geranylgeranyl diphosphate synthase, and three cytochrome P450 monooxygenases. We now describe a fourth cytochrome P450 monooxygenase gene (P450-4). Gas chromatography-mass spectrometry analysis of extracts of mycelia and culture fluid of a P450-4 knockout mutant identified ent-kaurene as the only intermediate of the GA pathway. Incubations with radiolabeled precursors showed that the metabolism of ent-kaurene, ent-kaurenol, and ent-kaurenal was blocked in the transformants, whereas ent-kaurenoic acid was metabolized efficiently to GA(4). The GA-deficient mutant strain SG139, which lacks the 30-kb GA biosynthesis gene cluster, converted ent-kaurene to ent-kaurenoic acid after transformation with P450-4. The B1-41a mutant, described as blocked between ent-kaurenal and ent-kaurenoic acid, was fully complemented by P450-4. There is a single nucleotide difference between the sequence of the B1-41a and wild-type P450-4 alleles at the 3' consensus sequence of intron 2 in the mutant, resulting in reduced levels of active protein due to a splicing defect in the mutant. These data suggest that P450-4 encodes a multifunctional ent-kaurene oxidase catalyzing all three oxidation steps between ent-kaurene and ent-kaurenoic acid. PMID- 11472928 TI - Effects of cell-bound microcystins on survival and feeding of Daphnia spp. AB - The influence of cell-bound microcystins on the survival time and feeding rates of six Daphnia clones belonging to five common species was studied. To do this, the effects of the microcystin-producing Microcystis strain PCC7806 and its mutant, which has been genetically engineered to knock out microcystin synthesis, were compared. Additionally, the relationship between microcystin ingestion rate by the Daphnia clones and Daphnia survival time was analyzed. Microcystins ingested with Microcystis cells were poisonous to all Daphnia clones tested. The median survival time of the animals was closely correlated to their microcystin ingestion rate. It was therefore suggested that differences in survival among Daphnia clones were due to variations in microcystin intake rather than due to differences in susceptibility to the toxins. The correlation between median survival time and microcystin ingestion rate could be described by a reciprocal power function. Feeding experiments showed that, independent of the occurrence of microcystins, cells of wild-type PCC7806 and its mutant are able to inhibit the feeding activity of Daphnia. Both variants of PCC7806 were thus ingested at low rates. In summary, our findings strongly suggest that (i) sensitivity to the toxic effect of cell-bound microcystins is typical for Daphnia spp., (ii) Daphnia spp. and clones may have a comparable sensitivity to microcystins ingested with food particles, (iii) Daphnia spp. may be unable to distinguish between microcystin-producing and -lacking cells, and (iv) the strength of the toxic effect can be predicted from the microcystin ingestion rate of the animals. PMID- 11472929 TI - Identification and functional characterization of CbaR, a MarR-like modulator of the cbaABC-encoded chlorobenzoate catabolism pathway. AB - In Comamonas testosteroni BR60 (formerly Alcaligenes sp. strain BR60), catabolism of the pollutant 3-chlorobenzoate (3CBA) is initiated by enzymes encoded by cbaABC, an operon found on composite transposon Tn5271 of plasmid pBRC60. The cbaABC gene product CbaABC converts 3CBA to protocatechuate (PCA) and 5-Cl-PCA, which are then metabolized by the chromosomal PCA meta (extradiol) ring fission pathway. In this study, cbaA was found to possess a sigma(70) type promoter. O(2) uptake experiments with whole cells and expression studies with cbaA-lacZ constructs showed that cbaABC was induced by 3CBA. Benzoate, which is not a substrate of the 3CBA pathway, was a gratuitous inducer, and CbaR, a MarR family repressor coded for by a divergently transcribed gene upstream of cbaABC, could modulate induction mediated by benzoate. Purified CbaR bound specifically to two regions of the cbaA promoter (P(cbaA)); site I, a high-affinity site, is between the transcriptional start point (position +1) and the start codon of cbaA, while site II, a lower-affinity site, overlaps position +1. 3CBA at concentrations as low as 40 microM interfered with binding to P(cbaA). PCA also interfered with binding, while benzoate only weakly disrupted binding. Unexpectedly, benzoate with a hydroxyl or carboxyl at position 3 improved CbaR binding. Data are also presented that suggest that an unidentified regulator is encoded on the chromosome that induces cbaABC in response to benzoate and 3CBA. PMID- 11472930 TI - Fate of the biological control agent Pseudomonas aureofaciens TX-1 after application to turfgrass. AB - The fate and impact of Pseudomonas aureofaciens TX-1 following application as a biocontrol agent for fungi in turfgrass were studied. The organism was applied with a modified irrigation system by using a preparation containing 1 x 10(6) P. aureofaciens TX-1 CFU ml(-1) about 100 times between May and August. We examined the impact of this repeated introduction of P. aureofaciens TX-1 (which is known to produce the antimicrobial compound phenazine-1-carboxylic acid) on the indigenous microbial community of the turfgrass system and on establishment of introduced bacteria in the soil system. A PCR primer-DNA hybridization probe combination was developed to accurately monitor the fate of P. aureofaciens TX-1 following application in irrigation water. To assess the impact of frequent P. aureofaciens TX-1 applications on the indigenous bacterial community, turfgrass canopy, thatch, and rhizosphere samples were obtained during the growing season from control and treated plots and subjected to DNA extraction procedures and denaturing gradient gel electrophoresis (DGGE). PCR amplification and hybridization of extracted DNA with the P. aureofaciens TX-1-specific primer probe combination revealed that P. aureofaciens TX-1 not only became established in the rhizosphere and thatch but also was capable of overwintering. Separation of PCR-amplified partial 16S rRNA genes by DGGE showed that the repeated application of P. aureofaciens TX-1 in irrigation water resulted in transient displacement of a leaf surface bacterial community member. There was no obvious alteration of any dominant members of the thatch and rhizosphere microbial communities. PMID- 11472931 TI - Phosphorus-31 nuclear magnetic resonance study of the effect of pentachlorophenol (PCP) on the physiologies of PCP-degrading microorganisms. AB - Free and agarose-encapsulated pentachlorophenol (PCP)-degrading Sphingomonas sp. isolates UG25 and UG30 were compared to Sphingomonas chlorophenolica ATCC 39723 with respect to the ability to degrade PCP. Pretreatment of the UG25 and UG30 strains with 50 microg of PCP per ml enabled the cells to subsequently degrade higher levels of this environmental pollutant. Similar treatment of ATCC 39723 cells had no effect on the level of PCP degraded by this strain. Phosphorus-31 nuclear magnetic resonance spectra of agarose-immobilized strains UG25 and UG30 grown in the absence of PCP showed that there was marked deenergization of the cells upon exposure to a nonlethal concentration of PCP (120 microg/ml). For example, no transmembrane pH gradient was observed, and the ATP levels were lower than the levels obtained in the absence of PCP. The transmembrane pH gradient and ATP levels were restored once the immobilized cells had almost completely degraded the PCP in the perfusion medium. PCP-pretreated cells, on the other hand, maintained their transmembrane pH gradient and ATP levels even in the presence of high levels of PCP. The ability of PCP-pretreated strain UG25 and UG30 cells to remain energized in the presence of PCP was shown to correlate with an altered membrane phospholipid profile; these cells had a higher concentration of cardiolipin than cells cultured in the absence of PCP. Strain ATCC 39723, which did not degrade higher levels of PCP after PCP pretreatment, did not show this response. PMID- 11472932 TI - Effect of primers hybridizing to different evolutionarily conserved regions of the small-subunit rRNA gene in PCR-based microbial community analyses and genetic profiling. AB - Genetic profiling techniques of microbial communities based on PCR-amplified signature genes, such as denaturing gradient gel electrophoresis or single-strand conformation polymorphism (SSCP) analysis, are normally done with PCR products of less than 500-bp. The most common target for diversity analysis, the small subunit rRNA genes, however, are larger, and thus, only partial sequences can be analyzed. Here, we compared the results obtained by PCR targeting different variable (V) regions (V2 and V3, V4 and V5, and V6 to V8) of the bacterial 16S rRNA gene with primers hybridizing to evolutionarily conserved flanking regions. SSCP analysis of single-stranded PCR products generated from 13 different bacterial species showed fewer bands with products containing V4-V5 (average, 1.7 bands per organism) than with V2-V3 (2.2 bands) and V6-V8 (2.3 bands). We found that the additional bands (>1 per organism) were caused by intraspecies operon heterogeneities or by more than one conformation of the same sequence. Community profiles, generated by PCR-SSCP from bacterial-cell consortia extracted from rhizospheres of field-grown maize (Zea mays), were analyzed by cloning and sequencing of the dominant bands. A total of 48 sequences could be attributed to 34 different strains from 10 taxonomical groups. Independent of the primer pairs, we found proteobacteria (alpha, beta, and gamma subgroups) and members of the genus Paenibacillus (low G+C gram-positive) to be the dominant organisms. Other groups, however, were only detected with single primer pairs. This study gives an example of how much the selection of different variable regions combined with different specificities of the flanking "universal" primers can affect a PCR based microbial community analysis. PMID- 11472933 TI - Sequence analysis and molecular characterization of the Lactococcus lactis temperate bacteriophage BK5-T. AB - The Lactococcus lactis temperate bacteriophage BK5-T is one of twelve type phages that define L. lactis phage species. This paper describes the nucleotide sequence and analysis of a 21-kbp region of the BK5-T genome and completes the nucleotide sequence of the genome of this phage. The 40,003-nucleotide linear genome encodes 63 open reading frames. Sequence runoff experiments showed that the cohesive ends of the BK5-T genome contained a 12-bp 3' single-stranded overhang with the sequence 5'-CACACACATAGG-3'. Two major BK5-T structural proteins, of approximately 30 and 20 kDa, were identified, and N-terminal sequence analysis determined that they were encoded by orf7 and orf12, respectively. A 169-bp fragment containing a 37-bp direct repeat and several smaller repeat sequences conferred resistance to BK5-T infection when introduced in trans to the host cell and is likely a part of the BK5-T origin of replication (ori). PMID- 11472934 TI - Molecular cloning, nucleotide sequence, and expression of genes encoding a polycyclic aromatic ring dioxygenase from Mycobacterium sp. strain PYR-1. AB - Mycobacterium sp. strain PYR-1 degrades high-molecular-weight polycyclic hydrocarbons (PAHs) primarily through the introduction of both atoms of molecular oxygen by a dioxygenase. To clone the dioxygenase genes involved in PAH degradation, two-dimensional (2D) gel electrophoresis of PAH-induced proteins from cultures of Mycobacterium sp. strain PYR-1 was used to detect proteins that increased after phenanthrene, dibenzothiophene, and pyrene exposure. Comparison of proteins from induced and uninduced cultures on 2D gels indicated that at least six major proteins were expressed (105, 81, 52, 50, 43, and 13 kDa). The N terminal sequence of the 50-kDa protein was similar to those of other dioxygenases. A digoxigenin-labeled oligonucleotide probe designed from this protein sequence was used to screen dioxygenase-positive clones from a genomic library of Mycobacterium sp. strain PYR-1. Three clones, each containing a 5,288 bp DNA insert with three genes of the dioxygenase system, were obtained. The genes in the DNA insert, from the 5' to the 3' direction, were a dehydrogenase, the dioxygenase small (beta)-subunit, and the dioxygenase large (alpha)-subunit genes, arranged in a sequence different from those of genes encoding other bacterial dioxygenase systems. Phylogenetic analysis showed that the large alpha subunit did not cluster with most of the known alpha-subunit sequences but rather with three newly described alpha subunits of dioxygenases from Rhodococcus spp. and Nocardioides spp. The genes from Mycobacterium sp. strain PYR-1 were subcloned and overexpressed in Escherichia coli with the pBAD/ThioFusion system. The functionality of the genes for PAH degradation was confirmed in a phagemid clone containing all three genes, as well as in plasmid subclones containing the two genes encoding the dioxygenase subunits. PMID- 11472935 TI - Methane oxidation and the competition for oxygen in the rice rhizosphere. AB - A mechanistic approach is presented to describe oxidation of the greenhouse gas methane in the rice rhizosphere of flooded paddies by obligate methanotrophic bacteria. In flooded rice paddies these methanotrophs compete for available O(2) with other types of bacteria. Soil incubation studies and most-probable-number (MPN) counts of oxygen consumers show that microbial oxygen consumption rates were dominated by heterotrophic and methanotrophic respiration. MPN counts of methanotrophs showed large spatial and temporal variability. The most abundant methanotrophs (a Methylocystis sp.) and heterotrophs (a Pseudomonas sp. and a Rhodococcus sp.) were isolated and characterized. Growth dynamics of these bacteria under carbon and oxygen limitations are presented. Theoretical calculations based on measured growth dynamics show that methanotrophs were only able to outcompete heterotrophs at low oxygen concentrations (frequently < 5 microM). The oxygen concentration at which methanotrophs won the competition from heterotrophs did not depend on methane concentration, but it was highly affected by organic carbon concentrations in the paddy soil. Methane oxidation was severely inhibited at high acetate concentrations. This is in accordance with competition experiments between Pseudomonas spp. and Methylocystis spp. carried out at different oxygen and carbon concentrations. Likely, methane oxidation mainly occurs at microaerophilic and low-acetate conditions and thus not directly at the root surface. Acetate and oxygen concentrations in the rice rhizosphere are in the critical range for methane oxidation, and a high variability in methane oxidation rates is thus expected. PMID- 11472936 TI - Region-specific transcriptional activity in the genome of Streptomyces coelicolor A3(2). AB - Transcriptional analysis of microbial genomes is an important component of functional genomics. Strategies such as hybridization of labeled total RNA against ordered clone libraries or differential-display approaches have already been carried out to identify expressed genes. We describe here an additional method which applies subtractive hybridization between genome-specific DNA and total RNA followed by a PCR approach to identify expressed microbial genes. With the new strategy, the expression of genes in the terminal regions of the linear Streptomyces coelicolor A3(2) chromosome and the accessory linear plasmid SCP1 was analyzed. The results indicate that the method is useful for the identification of expressed genes in actinomycetes and other microbial systems. We demonstrate for the first time that at least 24 genes in the chromosome end regions (silent regions) of S. coelicolor are actively expressed. In addition, several expressed SCP1 genes were identified, including a gene which shows high similarity to microbial dnaN genes and which seems to play a role in SCP1 maintenance. PMID- 11472937 TI - The phosphinomethylmalate isomerase gene pmi, encoding an aconitase-like enzyme, is involved in the synthesis of phosphinothricin tripeptide in Streptomyces viridochromogenes. AB - Streptomyces viridochromogenes Tu494 produces the antibiotic phosphinothricin tripeptide (PTT). In the postulated biosynthetic pathway, one reaction, the isomerization of phosphinomethylmalate, resembles the aconitase reaction of the tricarboxylic acid (TCA) cycle. It was speculated that this reaction is carried out by the corresponding enzyme of the primary metabolism (C. J. Thompson and H. Seto, p. 197-222, in L. C. Vining and C. Stuttard, ed., Genetics and Biochemistry of Antibiotic Production, 1995). However, in addition to the TCA cycle aconitase gene, a gene encoding an aconitase-like protein (the phosphinomethylmalate isomerase gene, pmi) was identified in the PTT biosynthetic gene cluster by Southern hybridization experiments, using oligonucleotides which were derived from conserved amino acid sequences of aconitases. The deduced protein revealed high similarity to aconitases from plants, bacteria, and fungi and to iron regulatory proteins from eucaryotes. Pmi and the S. viridochromogenes TCA cycle aconitase, AcnA, have 52% identity. By gene insertion mutagenesis, a pmi mutant (Mapra1) was generated. The mutant failed to produce PTT, indicating the inability of AcnA to carry out the secondary-metabolism reaction. A His-tagged protein (Hispmi*) was heterologously produced in Streptomyces lividans. The purified protein showed no standard aconitase activity with citrate as a substrate, and the corresponding gene was not able to complement an acnA mutant. This indicates that Pmi and AcnA are highly specific for their respective enzymatic reactions. PMID- 11472938 TI - Degradation of chlorinated dibenzofurans and dibenzo-p-dioxins by two types of bacteria having angular dioxygenases with different features. AB - Two kinds of bacteria having different-structured angular dioxygenases-a dibenzofuran (DF)-utilizing bacterium, Terrabacter sp. strain DBF63, and a carbazole (CAR)-utilizing bacterium, Pseudomonas sp. strain CA10-were investigated for their ability to degrade some chlorinated dibenzofurans (CDFs) and chlorinated dibenzo-p-dioxins (CDDs) (or, together, CDF/Ds) using either wild type strains or recombinant Escherichia coli strains. First, it was shown that CAR 1,9a-dioxygenase (CARDO) catalyzed angular dioxygenation of all mono- to triCDF/Ds investigated in this study, but DF 4,4a-dioxygenase (DFDO) did not degrade 2,7-diCDD. Secondly, degradation of CDF/Ds by the sets of three enzymes (angular dioxygenase, extradiol dioxygenase, and meta-cleavage compound hydrolase) was examined, showing that these enzymes in both strains were able to convert 2-CDF to 5-chlorosalicylic acid but not other tested substrates to the corresponding chlorosalicylic acid (CSA) or chlorocatechol (CC). Finally, we tested the potential of both wild-type strains for cooxidation of CDF/Ds and demonstrated that both strains degraded 2-CDF, 2-CDD, and 2,3-diCDD to the corresponding CSA and CC. We investigated the sites for the attack of angular dioxygenases in each CDF/D congener, suggesting the possibility that the angular dioxygenation of 2-CDF, 2-CDD, 2,3-diCDD, and 1,2,3-triCDD (10 ppm each) by both DFDO and CARDO occurred mainly on the nonsubstituted aromatic nuclei. PMID- 11472939 TI - Distribution of archaea in a black smoker chimney structure. AB - Archaeal community structures in microhabitats in a deep-sea hydrothermal vent chimney structure were evaluated through the combined use of culture-independent molecular analyses and enrichment culture methods. A black smoker chimney was obtained from the PACMANUS site in the Manus Basin near Papua New Guinea, and subsamples were obtained from vertical and horizontal sections. The elemental composition of the chimney was analyzed in different subsamples by scanning electron microscopy and energy-dispersive X-ray spectroscopy, indicating that zinc and sulfur were major components while an increased amount of elemental oxygen in exterior materials represented the presence of oxidized materials on the outer surface of the chimney. Terminal restriction fragment length polymorphism analysis revealed that a shift in archaeal ribotype structure occurred in the chimney structure. Through sequencing of ribosomal DNA (rDNA) clones from archaeal rDNA clone libraries, it was demonstrated that the archaeal communities in the chimney structure consisted for the most part of hyperthermophilic members and extreme halophiles and that the distribution of such extremophiles in different microhabitats of the chimney varied. The results of the culture-dependent analysis supported in part the view that changes in archaeal community structures in these microhabitats are associated with the geochemical and physical dynamics in the black smoker chimney. PMID- 11472940 TI - Biochemical characterization of ochratoxin A-producing strains of the genus Penicillium. AB - In order to explore the biochemical scope of ochratoxin A-producing penicillia, we screened 48 Penicillium verrucosum isolates for the production of secondary metabolites. Fungal metabolites were analyzed by high-pressure liquid or gas chromatography coupled to diode array detection or mass spectrometry. The following metabolites were identified: ochratoxins A and B, citrinin, verrucolones, verrucines, anacines, sclerotigenin, lumpidin, fumiquinazolines, alantrypinones, daldinin D, dipodazine, penigequinolines A and B, 2-pentanone, and 2-methyl-isoborneol. By use of average linking clustering based on binary (nonvolatile) metabolite data, the 48 isolates could be grouped into two large and clearly separated groups and a small outlying group of four non-ochratoxin producing isolates. The largest group, containing 24 isolates, mainly originating from plant sources, included the type culture of P. verrucosum. These isolates produced ochratoxin A, verrucolones, citrinin, and verrucines and had a characteristic dark brown reverse color on yeast extract-sucrose agar medium. Almost all of a group of 20 isolates mainly originating from cheese and meat products had a pale cream reverse color on yeast extract-sucrose agar medium and produced ochratoxin A, verrucolones, anacines, and sclerotigenin. This group included the former type culture of P. nordicum. We also found that P. verrucosum isolates and three P. nordicum isolates incorporated phenylalanine into verrucine and lumpidin metabolites, a finding which could explain why those isolates produced relatively lower levels of ochratoxins than did most isolates of P. nordicum. PMID- 11472941 TI - Purification and characterization of pyranose oxidase from the white rot fungus Trametes multicolor. AB - We purified an intracellular pyranose oxidase from mycelial extracts of the white rot fungus Trametes multicolor by using ammonium sulfate fractionation, hydrophobic interaction, ion-exchange chromatography, and gel filtration. The native enzyme has a molecular mass of 270 kDa as determined by equilibrium ultracentrifugation and is composed of four identical 68-kDa subunits as determined by matrix-assisted laser desorption ionization mass spectrometry. Each subunit contains one covalently bound flavin adenine dinucleotide as its prosthetic group. The enzyme oxidizes several aldopyranoses specifically at position C-2, and its preferred electron donor substrates are D-glucose, D xylose, and L-sorbose. During this oxidation reaction electrons are transferred to oxygen, yielding hydrogen peroxide. In addition, the enzyme catalyzes the two electron reduction of 1,4-benzoquinone, several substituted benzoquinones, and 2,6-dichloroindophenol, as well as the one-electron reduction of the ABTS [2,2' azinobis(3-ethylbenzthiazolinesulfonic acid)] cation radical. As judged by the catalytic efficiencies (k(cat)/K(m)), some of these quinone electron acceptors are much better substrates for pyranose oxidase than oxygen. The optimum pH of the pyranose oxidase-catalyzed reaction depends strongly on the electron acceptor employed and varies from 4 to 8. It has been proposed that the main metabolic function of pyranose oxidase is as a constituent of the ligninolytic system of white rot fungi that provides peroxidases with H(2)O(2). An additional function could be reduction of quinones, key intermediates that are formed during mineralization of lignin. PMID- 11472942 TI - Rapid evolution of novel traits in microorganisms. AB - The use of natural microorganisms in biotransformations is frequently constrained by their limited tolerance to the high concentrations of metabolites and solvents required for effective industrial production. In many cases, more robust strains have to be generated by random mutagenesis and selection. This process of directed evolution can be accelerated in mutator strains, which carry defects in one or more of their DNA repair genes. However, in order to use mutator strains, it is essential to restore the normal low mutation rate of the selected organisms immediately after selection to prevent the accumulation of undesirable spontaneous mutations. To enable this process, we constructed temperature sensitive plasmids that temporarily increase the mutation frequency of their hosts by 20- to 4,000-fold. Under appropriate selection pressure, microorganisms transformed with mutator plasmids can be quickly evolved to exhibit new, complex traits. By using this approach, we were able to increase the tolerance of three bacterial strains to dimethylformamide by 10 to 20 g/liter during only two subsequent transfers. Subsequently, the evolved strains were returned to their normal low mutation rate by curing the cells of the mutator plasmids. Our results demonstrate a new and efficient method for rapid strain improvement based on in vivo mutagenesis. PMID- 11472943 TI - L-pyroglutamate spontaneously formed from L-glutamate inhibits growth of the hyperthermophilic archaeon Sulfolobus solfataricus. AB - Identification of physiological and environmental factors that limit efficient growth of hyperthermophiles is important for practical application of these organisms to the production of useful enzymes or metabolites. During fed-batch cultivation of Sulfolobus solfataricus in medium containing L-glutamate, we observed formation of L-pyroglutamic acid (PGA). PGA formed spontaneously from L glutamate under culture conditions (78 degrees C and pH 3.0), and the PGA formation rate was much higher at an acidic or alkaline pH than at neutral pH. It was also found that PGA is a potent inhibitor of S. solfataricus growth. The cell growth rate was reduced by one-half by the presence of 5.1 mM PGA, and no growth was observed in the presence of 15.5 mM PGA. On the other hand, the inhibitory effect of PGA on cell growth was alleviated by addition of L-glutamate or L aspartate to the medium. PGA was also produced from the L-glutamate in yeast extract; the PGA content increased to 8.5% (wt/wt) after 80 h of incubation of a yeast extract solution at 78 degrees C and pH 3.0. In medium supplemented with yeast extract, cell growth was optimal in the presence of 3.0 g of yeast extract per liter, and higher yeast extract concentrations resulted in reduced cell yields. The extents of cell growth inhibition at yeast extract concentrations above the optimal concentration were correlated with the PGA concentration in the culture broth. Although other structural analogues of L-glutamate, such as L methionine sulfoxide, glutaric acid, succinic acid, and L-glutamic acid gamma methyl ester, also inhibited the growth of S. solfataricus, the greatest cell growth inhibition was observed with PGA. We also observed that unlike other glutamate analogues, N-acetyl-L-glutamate enhanced the growth of S. solfataricus. This compound was stable under cell culture conditions, and replacement of L glutamate with N-acetyl-L-glutamate in the medium resulted in increased cell density. PMID- 11472944 TI - Specific detection of Bradyrhizobium and Rhizobium strains colonizing rice (Oryza sativa) roots by 16S-23S ribosomal DNA intergenic spacer-targeted PCR. AB - In addition to forming symbiotic nodules on legumes, rhizobial strains are members of soil or rhizosphere communities or occur as endophytes, e.g., in rice. Two rhizobial strains which have been isolated from root nodules of the aquatic legumes Aeschynomene fluminensis (IRBG271) and Sesbania aculeata (IRBG74) were previously found to promote rice growth. In addition to analyzing their phylogenetic positions, we assessed the suitability of the 16S-23S ribosomal DNA (rDNA) intergenic spacer (IGS) sequences for the differentiation of closely related rhizobial taxa and for the development of PCR protocols allowing the specific detection of strains in the environment. 16S rDNA sequence analysis (sequence identity, 99%) and phylogenetic analysis of IGS sequences showed that strain IRBG271 was related to but distinct from Bradyrhizobium elkanii. Rhizobium sp. (Sesbania) strain IRBG74 was located in the Rhizobium-Agrobacterium cluster as a novel lineage according to phylogenetic 16S rDNA analysis (96.8 to 98.9% sequence identity with Agrobacterium tumefaciens; emended name, Rhizobium radiobacter). Strain IRBG74 harbored four copies of rRNA operons whose IGS sequences varied only slightly (2 to 9 nucleotides). The IGS sequence analyses allowed intraspecies differentiation, especially in the genus Bradyrhizobium, as illustrated here for strains of Bradyrhizobium japonicum, B. elkanii, Bradyrhizobium liaoningense, and Bradyrhizobium sp. (Chamaecytisus) strain BTA-1. It also clearly differentiated fast-growing rhizobial species and strains, albeit with lower statistical significance. Moreover, the high sequence variability allowed the development of highly specific IGS-targeted nested-PCR assays. Strains IRBG74 and IRBG271 were specifically detected in complex DNA mixtures of numerous related bacteria and in the DNA of roots of gnotobiotically cultured or even of soil-grown rice plants after inoculation. Thus, IGS sequence analysis is an attractive technique for both microbial ecology and systematics. PMID- 11472945 TI - Bacillus spore inactivation methods affect detection assays. AB - Detection of biological weapons is a primary concern in force protection, treaty verification, and safeguarding civilian populations against domestic terrorism. One great concern is the detection of Bacillus anthracis, the causative agent of anthrax. Assays for detection in the laboratory often employ inactivated preparations of spores or nonpathogenic simulants. This study uses several common biodetection platforms to detect B. anthracis spores that have been inactivated by two methods and compares those data to detection of spores that have not been inactivated. The data demonstrate that inactivation methods can affect the sensitivity of nucleic acid- and antibody-based assays for the detection of B. anthracis spores. These effects should be taken into consideration when comparing laboratory results to data collected and assayed during field deployment. PMID- 11472946 TI - Enrichment of high-affinity CO oxidizers in Maine forest soil. AB - Carboxydotrophic activity in forest soils was enriched by incubation in a flowthrough system with elevated concentrations of headspace CO (40 to 400 ppm). CO uptake increased substantially over time, while the apparent K(m) ((app)K(m)) for uptake remained similar to that of unenriched soils (<10 to 20 ppm). Carboxydotrophic activity was transferred to and further enriched in sterile sand and forest soil. The (app)K(m)s for secondary and tertiary enrichments remained similar to values for unenriched soils. CO uptake by enriched soil and freshly collected forest soil was inhibited at headspace CO concentrations greater than about 1%. A novel isolate, COX1, obtained from the enrichments was inhibited similarly. However, in contrast to extant carboxydotrophs, COX1 consumed CO with an (app)K(m) of about 15 ppm, a value comparable to that of fresh soils. Phylogenetic analysis based on approximately 1,200 bp of its 16S rRNA gene sequence suggested that the isolate is an alpha-proteobacterium most closely related to the genera Pseudaminobacter, Aminobacter, and Chelatobacter (98.1 to 98.3% sequence identity). PMID- 11472947 TI - Bacterial species determination from DNA-DNA hybridization by using genome fragments and DNA microarrays. AB - Whole genomic DNA-DNA hybridization has been a cornerstone of bacterial species determination but is not widely used because it is not easily implemented. We have developed a method based on random genome fragments and DNA microarray technology that overcomes the disadvantages of whole-genome DNA-DNA hybridization. Reference genomes of four fluorescent Pseudomonas species were fragmented, and 60 to 96 genome fragments of approximately 1 kb from each strain were spotted on microarrays. Genomes from 12 well-characterized fluorescent Pseudomonas strains were labeled with Cy dyes and hybridized to the arrays. Cluster analysis of the hybridization profiles revealed taxonomic relationships between bacterial strains tested at species to strain level resolution, suggesting that this approach is useful for the identification of bacteria as well as determining the genetic distance among bacteria. Since arrays can contain thousands of DNA spots, a single array has the potential for broad identification capacity. In addition, the method does not require laborious cross-hybridizations and can provide an open database of hybridization profiles, avoiding the limitations of traditional DNA-DNA hybridization. PMID- 11472948 TI - Cluster structure of anaerobic aggregates of an expanded granular sludge bed reactor. AB - The metabolic properties and ultrastructure of mesophilic aggregates from a full scale expanded granular sludge bed reactor treating brewery wastewater are described. The aggregates had a very high methanogenic activity on acetate (17.19 mmol of CH(4)/g of volatile suspended solids [VSS].day or 1.1 g of CH(4) chemical oxygen demand/g of VSS.day). Fluorescent in situ hybridization using 16S rRNA probes of crushed granules showed that 70 and 30% of the cells belonged to the archaebacterial and eubacterial domains, respectively. The spherical aggregates were black but contained numerous whitish spots on their surfaces. Cross sectioning these aggregates revealed that the white spots appeared to be white clusters embedded in a black matrix. The white clusters were found to develop simultaneously with the increase in diameter. Energy-dispersed X-ray analysis and back-scattered electron microscopy showed that the whitish clusters contained mainly organic matter and no inorganic calcium precipitates. The white clusters had a higher density than the black matrix, as evidenced by the denser cell arrangement observed by high-magnification electron microscopy and the significantly higher effective diffusion coefficient determined by nuclear magnetic resonance imaging. High-magnification electron microscopy indicated a segregation of acetate-utilizing methanogens (Methanosaeta spp.) in the white clusters from syntrophic species and hydrogenotrophic methanogens (Methanobacterium-like and Methanospirillum-like organisms) in the black matrix. A number of physical and microbial ecology reasons for the observed structure are proposed, including the advantage of segregation for high-rate degradation of syntrophic substrates. PMID- 11472949 TI - Induction of a mitosis delay and cell lysis by high-level secretion of mouse alpha-amylase from Saccharomyces cerevisiae. AB - Some foreign proteins are produced in yeast in a cell cycle-dependent manner, but the cause of the cell cycle dependency is unknown. In this study, we found that Saccharomyces cerevisiae cells secreting high levels of mouse alpha-amylase have elongated buds and are delayed in cell cycle completion in mitosis. The delayed cell mitosis suggests that critical events during exit from mitosis might be disturbed. We found that the activities of PP2A (protein phosphatase 2A) and MPF (maturation-promoting factor) were reduced in alpha-amylase-oversecreting cells and that these cells showed a reduced level of assembly checkpoint protein Cdc55, compared to the accumulation in wild-type cells. MPF inactivation is due to inhibitory phosphorylation on Cdc28, as a cdc28 mutant which lacks an inhibitory phosphorylation site on Cdc28 prevents MPF inactivation and prevents the defective bud morphology induced by overproduction of alpha-amylase. Our data also suggest that high levels of alpha-amylase may downregulate PPH22, leading to cell lysis. In conclusion, overproduction of heterologous alpha-amylase in S. cerevisiae results in a negative regulation of PP2A, which causes mitotic delay and leads to cell lysis. PMID- 11472950 TI - Characterization of a granulovirus isolated from Epinotia aporema Wals. (Lepidoptera: Tortricidae) larvae. AB - A granulovirus (GV) isolated from Epinotia aporema (Lepidoptera: Tortricidae)-a major soybean pest-was studied in terms of its main morphological, biochemical, and biological properties. The ovoidal occlusion bodies were 466 by 296 nm in size, and their most prominent protein had an apparent molecular mass of 29 kDa. Its amino-terminal sequence was remarkably homologous to that of the granulins of other GVs. The DNA genome size was estimated to be 120 kbp. The high specificity and pathogenicity of this newly described granulovirus (EpapGV) indicate that it is indeed a good candidate for the biological control of this pest. PMID- 11472951 TI - Purification and characterization of a vulnificolysin-like cytolysin produced by Vibrio tubiashii. AB - An extracellular cytolysin from Vibrio tubiashii was purified by sequential hydrophobic interaction chromatography with phenyl-Sepharose CL-4B and gel filtration with Sephacryl S-200. This protein is sensitive to heat and proteases, is inhibited by cholesterol, and has a molecular weight of 59,000 and an isoelectric point of 5.3. In addition to lysing various erythrocytes, it is cytolytic and/or cytotoxic to Chinese hamster ovary cells, Caco-2 cells, and Atlantic menhaden liver cells in tissue culture. Lysis of erythrocytes occurs by a multihit process that is dependent on temperature and pH. Twelve of the first 17 N-terminal amino acid residues (Asp-Asp-Tyr-Val-Pro-Val-Val-Glu-Lys-Val-Tyr Tyr-Ile-Thr-Ser-Ser-Lys) are identical to those of the Vibrio vulnificus cytolysin. PMID- 11472952 TI - Effectiveness of germicidal UV radiation for reducing fungal contamination within air-handling units. AB - Levels of fungi growing on insulation within air-handling units (AHUs) in an office building and levels of airborne fungi within AHUs were measured before the use of germicidal UV light and again after 4 months of operation. The fungal levels following UV operation were significantly lower than the levels in control AHUs. PMID- 11472953 TI - Transformation of 2,2'-bimorphine to the novel compounds 10-alpha-S-monohydroxy 2,2'-bimorphine and 10,10'-alpha,alpha'-S,S'-dihydroxy-2,2'-bimorphine by Cylindrocarpon didymum. AB - Whole-cell suspensions of Cylindrocarpon didymum were observed to transform 2,2' bimorphine to the compounds 10-alpha-S-monohydroxy-2,2'-bimorphine and 10,10' alpha,alpha'-S,S'-dihydroxy-2,2'-bimorphine. Mass spectrometry and (1)H nuclear magnetic resonance spectroscopy confirmed the identities of these new morphine alkaloids. PMID- 11472954 TI - Utilization of the rpoB gene as a specific chromosomal marker for real-time PCR detection of Bacillus anthracis. AB - The potential use of Bacillus anthracis as a weapon of mass destruction poses a threat to humans, domesticated animals, and wildlife and necessitates the need for a rapid and highly specific detection assay. We have developed a real-time PCR-based assay for the specific detection of B. anthracis by taking advantage of the unique nucleotide sequence of the B. anthracis rpoB gene. Variable region 1 of the rpoB gene was sequenced from 36 Bacillus strains, including 16 B. anthracis strains and 20 other related bacilli, and four nucleotides specific for B. anthracis were identified. PCR primers were selected so that two B. anthracis specific nucleotides were at their 3' ends, whereas the remaining bases were specific to the probe region. This format permitted the PCR reactions to be performed on a LightCycler via fluorescence resonance energy transfer (FRET). The assay was found to be specific for 144 B. anthracis strains from different geographical locations and did not cross-react with other related bacilli (175 strains), with the exception of one strain. The PCR assay can be performed on isolated DNA as well as crude vegetative cell lysates in less than 1 h. Therefore, the rpoB-FRET assay could be used as a new chromosomal marker for rapid detection of B. anthracis. PMID- 11472955 TI - Isolation and regulation of Sinorhizobium meliloti 1021 loci induced by oxygen limitation. AB - Eleven Sinorhizobium meliloti 1021 loci whose expression was induced under low oxygen concentrations were identified in a collection of 5,000 strains carrying Tn5-1063 (luxAB) transcriptional reporter gene fusions. The 11 Tn5-1063-tagged loci were cloned and characterized. The dependence of the expression of the tagged loci on the FixL/FixJ oxygen-sensing two-component regulatory system was examined. Three of the loci were found to be dependent upon fixL and fixJ for their expression, while one locus showed a partial dependence. The remaining seven loci showed fixL- and fixJ-independent induction of expression in response to oxygen limitation. This suggests that in S. meliloti, additional regulatory system(s) exist that respond either directly or indirectly to oxygen limitation conditions. PMID- 11472956 TI - Horizontal transfer of a multi-drug resistance plasmid between coliform bacteria of human and bovine origin in a farm environment. AB - Multi-drug-resistant coliform bacteria were isolated from feces of cattle exposed to antimicrobial agents and humans associated with the animals. Isolates from both cattle and humans harbored an R plasmid of 65 kb (pTMS1) that may have been transferred between them due to selective antibiotic pressure in the farm environment. PMID- 11472957 TI - Use of an intergenic region in Pseudomonas syringae pv. syringae B728a for site directed genomic marking of bacterial strains for field experiments. AB - To construct differentially-marked derivatives of our model wild-type strain, Pseudomonas syringae pv. syringae B728a (a causal agent of bacterial brown spot disease in snap bean plants), for field experiments, we selected a site in the gacS-cysM intergenic region for site-directed insertion of antibiotic resistance marker cassettes. In each of three field experiments, population sizes of the site-directed chromosomally marked B728a derivatives in association with snap bean plants were not significantly different from that of the wild-type strain. Inserts of up to 7 kb of DNA in the intergenic region did not measurably affect fitness of B728a in the field. The site is useful for site-directed genomic insertions of single copies of genes of interest. PMID- 11472958 TI - Isolation and in vivo and in vitro antifungal activity of phenylacetic acid and sodium phenylacetate from Streptomyces humidus. AB - The antifungal substances SH-1 and SH-2 were isolated from Streptomyces humidus strain S5-55 cultures by various purification procedures and identified as phenylacetic acid and sodium phenylacetate, respectively, based on the nuclear magnetic resonance, electron ionization mass spectral, and inductively coupled plasma mass spectral data. SH-1 and SH-2 completely inhibited the growth of Pythium ultimum, Phytophthora capsici, Rhizoctonia solani, Saccharomyces cerevisiae, and Pseudomonas syringae pv. syringae at concentrations from 10 to 50 microg/ml. The two compounds were as effective as the commercial fungicide metalaxyl in inhibiting spore germination and hyphal growth of P. capsici. However, the in vivo control efficacies of the two antifungal compounds against P. capsici infection on pepper plants were similar to those of H(3)PO(3) and fosetyl-AI but less than that of metalaxyl. PMID- 11472959 TI - Purification and characterization of a dimethoate-degrading enzyme of Aspergillus niger ZHY256, isolated from sewage. AB - A dimethoate-degrading enzyme from Aspergillus niger ZHY256 was purified to homogeneity with a specific activity of 227.6 U/mg of protein. The molecular mass of the purified enzyme was estimated to be 66 kDa by gel filtration and 67 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point was found to be 5.4, and the enzyme activity was optimal at 50 degrees C and pH 7.0. The activity was inhibited by most of the metal ions and reagents, while it was induced by Cu(2+). The Michaelis constant (K(m)) and V(max) for dimethoate were 1.25 mM and 292 micromol min(-1) mg of protein(-1), respectively. PMID- 11472960 TI - Evidence for iron-dependent nitrate respiration in the dissimilatory iron reducing bacterium Geobacter metallireducens. AB - The dissimilatory iron-reducing bacterium Geobacter metallireducens was found to require iron at a concentration in excess of 50 microM for continuous cultivation on nitrate. Growth yield (approximately 3-fold), cytochrome c content (approximately 7-fold), and nitrate (approximately 4.5-fold) and nitrite (approximately 70-fold) reductase activities were all increased significantly when the growth medium was amended with 500 microM iron. PMID- 11472961 TI - Optimization of annealing temperature to reduce bias caused by a primer mismatch in multitemplate PCR. AB - To reduce PCR bias derived from a primer mismatch, the effect of the annealing temperature on the product ratio was investigated by denaturing gradient gel electrophoresis analysis of PCR products from a mixture of perfect-match and one mismatch templates. These templates were generated by PCR from Pediococcus acidilactici for one mismatch and Micrococcus luteus for the perfect match. PCRs showed that the bias was reduced at lower temperatures. An environmental sample was also examined. PMID- 11472962 TI - Recovery of Streptococcus iniae from diseased fish previously vaccinated with a streptococcus vaccine. AB - Streptococcus iniae was recovered from diseased rainbow trout (Oncorhynchus mykiss, Walbaum) previously vaccinated against streptococcosis. PCR and serological methods indicate the presence of a new serotype in the diseased fish. PMID- 11472963 TI - Identification of nucleotide sequences for the specific and rapid detection of Yersinia pestis. AB - Suppression subtractive hybridization, a cost-effective approach for targeting unique DNA, was used to identify a 41.7-kb Yersinia pestis-specific region. One primer pair designed from this region amplified PCR products from natural isolates of Y. pestis and produced no false positives for near neighbors, an important criterion for unambiguous bacterial identification. PMID- 11472964 TI - Antibiotic resistance in Salmonella enterica serovar Typhimurium exposed to microcin-producing Escherichia coli. AB - Microcin 24 is an antimicrobial peptide secreted by uropathogenic Escherichia coli. Secretion of microcin 24 provides an antibacterial defense mechanism for E. coli. In a plasmid-based system using transformed Salmonella enterica, we found that resistance to microcin 24 could be seen in concert with a multiple antibiotic resistance phenotype. This multidrug-resistant phenotype appeared when Salmonella was exposed to an E. coli strain expressing microcin 24. Therefore, it appears that multidrug-resistant Salmonella can arise as a result of an insult from other pathogenic bacteria. PMID- 11472965 TI - Sinorhizobium meliloti cells require biotin and either cobalt or methionine for growth. AB - Sinorhizobium meliloti is usually cultured in rich media containing yeast extract. It has been suggested that some components of yeast extract are also required for growth in minimal medium. We tested 27 strains of this bacterium and found that none were able to grow in minimal medium when methods to limit carryover of yeast extract were used during inoculation. By fractionation of yeast extract, two required growth factors were identified. Biotin was found to be absolutely required for growth, whereas previously the need for this vitamin was considered to be strain specific. All strains also required supplementation with cobalt or methionine, consistent with the requirement for a vitamin B(12) dependent homocysteine methyltransferase for methionine biosynthesis. PMID- 11472967 TI - Transforming growth factor-beta(1) overexpression in tumor necrosis factor-alpha receptor knockout mice induces fibroproliferative lung disease. AB - Tumor necrosis factor-alpha receptor knockout (TNF-alphaRKO) mice have homozygous deletions of the genes that code for both the 55- and 75-kD receptors. The mice are protected from the fibrogenic effects of bleomycin, silica, and inhaled asbestos. The asbestos-exposed animals exhibit reduced expression of other peptide growth factors such as transforming growth factor (TGF)-alpha, platelet derived growth factors, and TGF-beta. In normal animals, these and other cytokines are elaborated at high levels during the development of fibroproliferative lung disease, but there is little information available that has allowed investigators to establish the role of the individual growth factors in disease pathogenesis. Here, we show that overexpression of TGF-beta(1) by means of a replication-deficient adenovirus vector induces fibrogenesis in the lungs of the fibrogenic-resistant TNF-alphaRKO mice. The fibrogenic lesions developed in both the KO and background controls within 7 d, and both types of animals exhibited similar incorporation of bromodeoxyuridine. Interestingly, airway epithelial cell proliferation appeared to be suppressed, perhaps due to the presence of the TGF-beta(1), a well-known inhibitor of epithelial mitogenesis. Before these experiments, there was no information available that would provide a basis for predicting whether or not TGF-beta(1) expression induces fibroproliferative lung disease in fibrogenic-resistant TNF-alphaRKO mice, an increasingly popular animal model. PMID- 11472968 TI - Fetal lung liquid secretion: insights using the tools of inhibitors and genetic knock-out experiments. PMID- 11472969 TI - Chinks in the armor of the airway: Pseudomonas infection in the cystic fibrosis lung. PMID- 11472970 TI - Liquid and ion transport by fetal airway and lung epithelia of mice deficient in sodium-potassium-2-chloride transporter. AB - Chloride (Cl(-)) movement across fetal lung epithelia is thought to be mediated by the sodium-potassium-2-Cl(-) cotransporter NKCC1. We studied the role of NKCC1 in Cl(-) and liquid secretion in late-gestation NKCC-null (-/-) and littermate control fetal mouse lung. NKCC -/- mice had decreased lung water compared with littermate controls (wet/dry: control, 8.01 +/- 0.09; NKCC -/-, 7.06 +/- 0.14). Liquid secretion by 17-d NKCC -/- distal lung explants was similar to control explants. Bumetanide inhibited basal liquid secretion in control but not NKCC -/- explants (expansion over 48 h: control, 35 +/- 4%; NKCC -/- 46 +/- 7%). Treatment with 4,4'-diisothiocyanto-stilbene-2,2'-disulfonic acid (DIDS) decreased liquid secretion in both control and NKCC -/- explants. Basal transepithelial potential difference (PD) of control tracheal explants was higher than that of NKCC -/- (control, -13.7 +/- 0.5 mV; NKCC -/-, -11.6 +/- 0.6 mV). Amiloride (10(-)(4) M) inhibited basal PD to the same extent in control and NKCC -/- mice. Terbutaline stimulated hyperpolarization was less in NKCC -/- than in control tracheas (DeltaPD: control, -10.8 +/- 1.33 mV; NKCC -/-, -6.1 +/- 0.7 mV) and was inhibited by DIDS and acetazolamide in NKCC -/- but not wild-type explants. We conclude that NKCC is rate-limiting for transcellular Cl(-) transport, and that alternative anion transport mechanisms can sustain liquid production at near normal levels in the fetal NKCC -/- mouse lung. PMID- 11472971 TI - Salt-independent abnormality of antimicrobial activity in cystic fibrosis airway surface fluid. AB - The link between the genetic defect in cystic fibrosis (CF) and the recently described breach in pulmonary host defense has focused on the role of salt and water metabolism in the airways. Using a human bronchial xenograft model we demonstrate a salt-independent abnormality in bacterial killing in CF airway surface fluid (ASF). Biochemical characterization implicates the absence or dysfunction of a molecule critical to the constitution of normal bacterial killing. Our study suggests that CF transmembrane conductance regulator (CFTR) deficiency causes a primary abnormality in the composition of ASF that leads to a salt-independent defect in host defense. Importantly, this defect is corrected by adenovirus-mediated gene transfer of CFTR. PMID- 11472972 TI - Oral tolerance induction by type V collagen downregulates lung allograft rejection. AB - Immunization with specific proteins or peptides has been used to induce immunologic tolerance to allografts other than the lung. Recently, we have reported that the immune response to lung alloantigen also involves an immune response to type V collagen [col(V)]. The purpose of the current study was to determine if oral administration of col(V) to lung allograft recipients before transplantation downregulates acute rejection episodes. The data show that, compared with controls, col(V)-fed recipients had fewer polymorphonuclear cells and lymphocytes in allograft bronchoalveolar lavage fluid, and reduced rejection pathology. Data showing that col(V)- fed allograft recipients had diminished delayed-type hypersensitivity (DTH) responses to donor alloantigens suggest that feeding col(V) prevented allograft rejection by inducing tolerance to donor antigens. Systemic production of transforming growth factor (TGF)-beta, interleukin (IL)-4, or IL-10 has been reported to be a mechanism for oral tolerance-induced suppression of immune responses. Feeding col(V) induced upregulated production of TGF-beta, but not IL-4 or IL-10 in serum. Neutralizing TGF-beta recovered the DTH response to donor antigen in tolerant allograft recipients. Collectively, these data show that oral administration of col(V) is a novel approach to induce immunologic tolerance to lung allografts, and that TGF beta contributed to suppression of the rejection response. PMID- 11472973 TI - Decreased surfactant protein-B expression and surfactant dysfunction in a murine model of acute lung injury. AB - This study examines the relationships between inflammation, surfactant protein (SP) expression, surfactant function, and lung physiology in a murine model of acute lung injury (ALI). 129/J mice received aerosolized endotoxin lipopolysaccharide [LPS] daily for up to 96 h to simulate the cytokine release and acute inflammation of ALI. Lung elastance (E(L)) and resistance, lavage fluid cell counts, cytokine levels, phospholipid and protein content, and surfactant function were measured. Lavage and lung tissue SP content were determined by Western blot and immunohistochemistry, and tissue messenger RNA (mRNA) levels were assessed by Northern blot and in situ hybridization. Tumor necrosis factor alpha and neutrophil counts in bronchoalveolar lavage fluid increased within 2 h of LPS exposure, followed by increases in total protein, interleukin (IL)-1beta, IL-6, and interferon-gamma. E(L) increased within 24 h of LPS exposure and remained abnormal up to 96 h. SP-B protein and mRNA levels were decreased at 24, 48, and 96 h. By contrast, SP-A protein and mRNA levels and SP-C mRNA levels were not reduced. Surfactant dysfunction occurred coincident with changes in SP-B levels. This study demonstrates that lung dysfunction in mice with LPS-ALI corresponds closely with abnormal surfactant function and reduced SP-B expression. PMID- 11472974 TI - Aspergillus fumigatus-induced allergic airway inflammation alters surfactant homeostasis and lung function in BALB/c mice. AB - The differential regulation of pulmonary surfactant proteins (SPs) is demonstrated in a murine model of Aspergillus fumigatus (Af )-induced allergic airway inflammation and hyperresponsiveness. BALB/c mice were sensitized intraperitoneally and challenged intranasally with Af extract. Enzyme-linked immunosorbent assay analysis of serum immunoglobulin (Ig) levels in these mice showed markedly increased total IgE and Af-specific IgE and IgG1. This was associated with peribronchial/perivascular tissue inflammation, airway eosinophilia, and secretion of interleukin (IL)-4 and IL-5 into the bronchoalveolar lavage fluid (BALF). Functional analysis revealed that in comparison with nonsensitized mice, allergic sensitization and challenge resulted in significant increases in acetylcholine responsiveness. To analyze levels of SPs, the cell-free supernate of the BALF was further fractionated by high-speed (20,000 x g) centrifugation. After sensitization and challenges, the pellet (large-aggregate fraction) showed a selective downregulation of hydrophobic SPs SP-B and SP-C by 50%. This reduction was reflected by commensurate decreases in SP-B and SP-C messenger RNA (mRNA) expression of the lung tissue of these animals. In contrast, there was a 9-fold increase in SP-D protein levels in the 20,000 x g supernate without changes in SP-D mRNA. The increased levels of SP-D showed a significant positive correlation with serum IgE (r = 0.85, P < 0.001). Tissue mRNA and protein levels of SP-A in either the large- or the small aggregate fractions were unaffected. Our data indicate that allergic airway inflammation induces selective inhibition of hydrophobic SP synthesis accompanied by marked increases in the lung collectin SP-D protein content of BALF. These changes may contribute significantly to the pathophysiology of Af-induced allergic airway hyperresponsiveness. PMID- 11472975 TI - Elevated expression of surfactant proteins in newborn rats during adaptation to hyperoxia. AB - The mechanisms whereby lung adaptation to hyperoxia occurs in the newborn period are incompletely understood. Pulmonary surfactant has been implicated in lung protection against hyperoxic injury, and elevated expression of certain surfactant proteins occurs in lungs of adult rats during adaptation to sublethal oxygen (85% O(2)). Here we report that newborn rats, which can adapt to even higher levels of hyperoxia (100% O(2)) than do adult rats, manifest changes in the lung surfactant proteins (SP), especially SP-A and SP-D. In newborn rats exposed to hyperoxia on Days 3 through 10 of life, lung messenger RNAs (mRNAs) for SP-A and SP-B gradually and progressively increased, relative to levels in age-matched, air-exposed newborns, over this 8-d period. By contrast, SP-C and SP D mRNAs were maximally increased relative to values in simultaneously air-exposed control rats after 4 d of exposure. Lung mRNA for CC-10, a protein specific for Clara cells, was greater in hyperoxia-exposed rats than in air-exposed control rats on Day 4 of exposure, but not on other days. Lung mRNA for thyroid transcription factor (TTF)-1 was marginally increased on Days 1, 2, 4, and 6, and significantly increased on Day 8. Both SP-A and SP-D proteins were increased in lung lavage samples taken from hyperoxia-exposed newborns, relative to those taken from air-exposed controls, with the greatest increases occurring on Days 6 and 8 of exposure. However, the patterns of increase of the proteins were not identical to those of the respective mRNAs. In situ hybridization studies demonstrated increases in SP-D, and to a lesser extent in SP-A, in peripheral lung tissues from oxygen-exposed newborns. Taken together, these data indicate that specific surfactant proteins are upregulated at both the pretranslational and post-translational levels in distal lung epithelium during adaptation to hyperoxia in the newborn rat. PMID- 11472976 TI - Activation of the TGF-beta/activin-Smad2 pathway during allergic airway inflammation. AB - Changes in the levels of transforming growth factor (TGF)-beta cytokines or receptors observed during the progression of several inflammatory and fibrotic disorders have been used to implicate these cytokines in the pathophysiology of these diseases. Although correlative, these studies were inconclusive because they were unable to demonstrate actual continuous TGF-beta-mediated signaling in the involved tissues. We reasoned that the phosphorylation state and subcellular localization of Smad2, the intracellular effector of TGF-beta/activin-mediated signaling, could be used as a marker of active signaling mediated by these cytokines in situ. We therefore used an experimental model of ovalbumin-induced allergic airway inflammation and were able to demonstrate a dramatic increase in the numbers of bronchial epithelial, alveolar, and infiltrating inflammatory cells expressing nuclear phosphorylated Smad2 within the allergen-challenged lungs. This was accompanied by strong upregulation of the activin receptor ALK 4/ActR-IB and redistribution of the TGF-beta responsive ALK-5/TbetaR-I. Although levels of TGF-beta1, TGF-beta2, and TGF-beta3 messenger RNA (mRNA) were marginally altered, the level of activin mRNA was strongly upregulated during the inflammatory response. Our data illustrate the usefulness of antiphosphorylated Smad antibodies in demonstrating active TGF- beta/activin-mediated signaling in vivo and strongly suggest that activin/Smad-mediated signaling could be a critical contributor in the pathophysiology of allergic pulmonary diseases. PMID- 11472977 TI - Naive and memory T cells migrate in comparable numbers through the normal rat lung: only effector T cells accumulate and proliferate in the lamina propria of the bronchi. AB - T cells reach the lung via the pulmonary and bronchial arteries that supply the alveolar and bronchial regions. Although these regions are differentially affected by T cell-mediated diseases, the migration of T-cell subsets in these two regions has not been studied. Naive, memory, and effector T cells were injected into congenic rats and traced in sections of normal lung. All three T cell subsets were found in large numbers in the alveolar region and exited again quickly. Only effector T cells accumulated in the lamina propria of the bronchi. Further, 72 h after injection 6% of the effector T cells still proliferated in the lung, whereas apoptotic effector T cells were only observed 1 h after injection (0.2%). Thus, not only effector and memory but also naive T cells continuously migrated through the lung. The preferential accumulation of effector T cells in the bronchial lamina propria may explain why some diseases preferentially affect the bronchial region. PMID- 11472978 TI - Polycyclic aromatic hydrocarbon diol epoxides increase cytosolic Ca(2+) of airway epithelial cells. AB - Polycyclic aromatic hydrocarbons (PAHs) increase cytosolic Ca(2+) concentration ([Ca(2+)](i)) in lymphocytes and mammary epithelial cells, but little is known regarding their effects on [Ca(2+)](i) in airway epithelium. We hypothesized that benzo[a]pyrene (BP) and/or anti-7,8-dihydroxy-9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene (BPDE), a carcinogenic BP metabolite, increases [Ca(2+)](i) in untransformed human small airway epithelial (SAE) cells and that their effects on [Ca(2+)](i) are directly proportional to carcinogenicity. SAE [Ca(2+)](i) was determined by a ratiometric digital Ca(2+) imaging system. BPDE increased SAE [Ca(2+)](i) within 20 s in media with high (1 mM) and low (10 nM) Ca(2+) at a threshold concentration of 0.2 nM. Elevation of [Ca(2+)](i) persisted longer with high Ca(2+). Neither BP nor solvent altered [Ca(2+)](i). Thapsigargin and inositol 1,4,5- phosphate receptor (InsP(3)R) antagonists inhibited this BPDE action with low Ca(2+). We conclude that BPDE but not BP increases [Ca(2+)](i) partly by mobilizing Ca(2+) from cytosolic stores through an InsP(3)R. The most potent carcinogenic PAH diol epoxide increased in SAE [Ca(2+)](i) at the lowest threshold concentration, suggesting that carcinogenicity is directly proportional to the action of PAHs on SAE [Ca(2+)](i). Short-term exposure to BPDE 36 to 48 h before the study rendered SAE cells less sensitive to BPDE, suggesting that BPDE may also induce persistent changes in Ca(2+) signaling pathways. PMID- 11472979 TI - Cytokine-mediated induction of the human elafin gene in pulmonary epithelial cells is regulated by nuclear factor-kappaB. AB - Elafin, a low molecular-weight proteinase inhibitor, is a member of the recently described trappin gene family. These proteins are thought to play important roles in the regulation of inflammation and are expressed in multiple epithelia. Elafin is found within the lung, and its expression can be induced by inflammatory mediators. The molecular mechanisms that mediate its induction are not understood. In this study we investigated the transcriptional regulation of the elafin gene in pulmonary epithelial cell lines. Transfection of elafin promoter constructs into the elafin-expressing pulmonary epithelial cell line A549 identified a number of positive-acting elements. Cytokine-mediated inducibility of the elafin gene promoter was shown to occur through a nuclear factor (NF) kappaB site present within the minimal promoter. This site was shown to bind to NF-kappaB proteins within nuclear extracts from cytokine stimulated cell lines as well as to in vitro-translated RelA. Cotransfection with both RelA and NF-kappaB inducing kinase induced reporter gene activation via this site, and mutagenesis experiments confirmed that it was crucial for induction of elafin gene activity. These results clearly identify a role for elafin in the inflammatory response of the airway epithelium to pathogenic insult and show that this response is mediated by an NF-kappaB site within the proximal promoter. PMID- 11472980 TI - Geldanamycin inhibits NF-kappaB activation and interleukin-8 gene expression in cultured human respiratory epithelium. AB - Geldanamycin is a benzoquinone ansamycin with multiple pharmacologic properties. Recent data demonstrated that geldanamycin conferred protection in an animal model of inflammation-associated acute lung injury. In the current study, we investigated the effects of geldanamycin on interleukin (IL)-8 gene expression and nuclear factor (NF)-kappaB activation. Geldanamycin inhibited tumor necrosis factor (TNF)-alpha-mediated IL-8 gene expression in A549 human respiratory epithelial cells as measured by enzyme-linked immunosorbent assay and Northern blot analyses. In cells transiently transfected with an IL-8 promoter-luciferase reporter plasmid, geldanamycin inhibited TNF-alpha-mediated luciferase activity. Geldanamycin inhibited TNF-alpha-mediated NF-kappaB activation as measured by electromobility shift assays and transient transfections with a NF-kappaB dependent luciferase reporter plasmid. In contrast, geldanamycin did not affect TNF-alpha-mediated degradation of the NF-kappaB inhibitory protein IkappaBalpha and did not block nuclear translocation of the NF-kappaB p65 subunit as measured by Western blot analyses. Geldanamycin added directly to nuclear extracts of TNF alpha-treated cells reduced the formation of the NF-kappaB/DNA complex. These results demonstrate that geldanamycin inhibits TNF-alpha-mediated IL-8 gene expression in A549 cells by inhibiting activation of the IL-8 promoter. The mechanism of inhibition involves inhibition of NF-kappaB activation, which is independent of IkappaBalpha degradation or p65 nuclear translocation. Geldanamycin appears to directly inhibit the ability of NF-kappaB to bind DNA. The observed in vitro effects could account, in part, for the anti-inflammatory properties of geldanamycin observed in vivo. PMID- 11472981 TI - Molecular and biochemical characterization of a CNP-sensitive guanylyl cyclase in bovine tracheal smooth muscle. AB - Muscarinic activation of bovine tracheal smooth muscle (BTSM) is involved in cyclic guanosine monophosphate (cGMP) production mediated through soluble (sGC) and membrane-bound (mGC) guanylyl cyclases. A muscarinic- and NaCl-sensitive mGC exists in BTSM regulated by muscarinic receptors coupled to G proteins. To identify the mGCs expressed in BTSM, reverse transcriptase/polymerase chain reaction (RT-PCR) from total RNA was performed using degenerate oligonucleotides for amplification of a region conserved among GC catalytic domains. Cloning of amplification products revealed that 76% of all BTSM GC transcripts corresponded to the sGC beta1 subunit and 24% to the B-type (C-type NP 1-22 [CNP]-sensitive) GC receptor. cGMP production by BTSM membrane and soluble fractions confirmed that sGC activity is 3-fold with respect to mGC activity. RT-PCR using specific oligonucleotides revealed that A (atrial NP-sensitive) and C (guanylin-sensitive) mGC subtypes are also expressed in BTSM. Stimulation of basal plasma membrane GC activity by CNP was higher than that by ANP, whereas guanylin showed no effect, indicating that CNP-sensitive guanylyl cyclase (GC-B) is the predominant functional BTSM mGC subtype. Strong adenosine triphosphate inhibition of CNP stimulated mGC activity supports the finding that the tracheal mGC isoform belongs to the natriuretic peptide-sensitive mGCs. Additionally, CNP was able to reverse the chloride inhibition of BTSM mGC activity, suggesting that this is a novel G protein-coupled GC-B receptor. PMID- 11472982 TI - Endogenously released endothelin-1 from human pulmonary artery smooth muscle promotes cellular proliferation: relevance to pathogenesis of pulmonary hypertension and vascular remodeling. AB - Endothelin (ET)-1 is a potent vasoconstrictor and comitogen/ proliferation factor for vascular smooth muscle (VSM). As such, it has been implicated in the vascular wall remodeling observed in pulmonary hypertension (PH). Although the endothelium is considered the main source of ET-1, it can be released by other cells including VSM and may mediate proliferation in an autocrine manner. We investigated this possibility using human pulmonary artery smooth-muscle (HPASM) cells. Serum stimulated the release of ET-1 from HPASM cells in a concentration dependent fashion and caused proliferation as determined by [(3)H]thymidine uptake and increase in cell number. Addition of an ET-A receptor antagonist (BQ123) or an inhibitor of ET-1 synthesis (phosphoramidon) reduced the proliferation induced by serum, confirming an autocrine role for ET-1. In addition, treatment of HPASM cells with two drug types used in the management of PH-cicaprost, a stable prostacyclin-mimetic; or diltiazem, a calcium-channel blocker-reduced ET-1 release from these cells. We conclude that ET-1 released from HPASM cells has an autocrine function in serum-induced proliferation, with important implications for the pathogenesis of human vascular remodeling. Drugs used in the treatment of PH may act, at least in part, by inhibiting this autocrine loop. PMID- 11472983 TI - Tumor necrosis factor-related apoptosis-inducing ligand and chemotherapy cooperate to induce apoptosis in mesothelioma cell lines. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can induce apoptosis in certain tumor cells. In addition, TRAIL and chemotherapy can act cooperatively, possibly as a result of chemotherapy-induced increases in expression of a TRAIL receptor, DR5. We used cell lines derived from a highly chemoresistant tumor, malignant mesothelioma, to learn whether TRAIL was effective alone or together with chemotherapy and whether cooperativity depended on increases in DR5 expression. TRAIL (codons 95-285) was expressed in a bacterial expression vector and purified by nickel affinity chromatography. TRAIL alone (25 to 500 ng/ml) had little effect on mesothelioma cells. TRAIL plus chemotherapy (doxorubicin, cis-platinum, etoposide, or gemcitabine) acted cooperatively to induce apoptosis in mesothelioma cells (M28, REN, VAMT, and MS 1). For example, in M28 cells treated for 18 h, apoptosis from TRAIL (100 ng/ml) plus doxorubicin (0.6 microg/ml; 71 +/- 11%) greatly exceeded that from TRAIL alone (21 +/- 8%) or from doxorubicin alone (6 +/- 2%) (means +/- standard deviation; P < 0.03). Mesothelioma cells treated with chemotherapy showed no change in DR5 protein by Western analysis or by immunocytochemistry. TRAIL plus chemotherapy was associated with an increase in mitochondrial cytochrome c release and mitochondrial depolarization. We conclude that TRAIL and chemotherapy act cooperatively to kill mesothelioma cell lines, not by increases in DR5 receptor but in association with mitochondrial amplification of apoptotic signals. PMID- 11472984 TI - Cloning of dexamethasone-induced transcript: a novel glucocorticoid-induced gene that is upregulated in emphysema. AB - To identify changes in gene expression associated with emphysema, we used differential display to compare RNA extracted from emphysematous lungs with that of unused donor tissues taken at the time of transplant. A differentially expressed sequence was identified corresponding to the 3' end of a novel human complementary DNA (cDNA) of unknown function. The human and mouse cDNA sequences were completed by 5' rapid amplification of cDNA ends. We have named it DEXI for dexamethasone-induced transcript. DEXI messenger RNA (mRNA) was upregulated 147% in emphysematous tissue compared with donor tissue. DEXI mRNA was also upregulated 230% by dexamethasone treatment of A549. The increase in expression of DEXI found in emphysema patients' tissues may be owing to their known treatment with corticosteroids. The human DEXI gene is intronless and the predicted open reading frame encodes a 95-residue acidic protein. Database searches revealed the presence of homologues only in mammals, and a human pseudogene. The protein has a predicted central transmembrane domain and a carboxy-terminal leucine zipper. The human mRNA has a single 1.3-kb transcript. We suggest that the increased expression of DEXI in emphysema may either be relevant to disease progression or be indicative of glucocorticoid responsiveness in treated patients. PMID- 11472985 TI - Cytokine profile of bronchoalveolar lavage-derived CD4(+), CD8(+), and gammadelta T cells in people with asthma after segmental allergen challenge. AB - T cell-derived cytokines play an important role in the pathogenesis of allergic asthma, but little is known about the cytokine profile of their different subsets. The aim of the present study was to investigate the cytokine production potential of CD4(+), CD8(+), or gammadelta(+) T cells derived from the bronchoalveolar space of mild atopic asthmatic subjects (n = 11) and nonatopic control subjects (n = 9) before and 24 h after segmental allergen challenge. The cytokine production was determined using the technique of intracellular cytokine detection by flow cytometry. Comparing asthmatic with control subjects we found no difference in the percentage of CD4(+), CD8(+), or gammadelta T cells in the bronchoalveolar lavage fluid before and after allergen challenge. Before allergen challenge the proportion of cells producing the cytokines interferon (IFN)-gamma, interleukin (IL)-2, IL-4, IL-5, and IL-13 was not different in CD4(+) and CD8(+) cells. The major difference between the groups was an increased percentage of positive-staining cells for the T helper-(Th)2-cytokines IL-5 and IL-13 in the gammadelta T-cell subset. After allergen challenge, all T-cell subsets revealed a decreased proportion of cells producing the Th1-type cytokines IFN-gamma and IL 2. The percentage of IL-4- and IL-5-positive cells did not change in all subsets, and there was a decreased proportion of IL-13- positive cells in the CD4(+) subset. These findings indicate an increased Th2-cytokine profile in gammadelta T cells. After allergen challenge, the dysbalance between Th1 and Th2 cytokines was further accentuated by a reduction in Th1 cytokine-producing T cells. PMID- 11472987 TI - An efficient algorithm for finding short approximate non-tandem repeats. AB - We study the problem of approximate non-tandem repeat extraction. Given a long subject string S of length N over a finite alphabet Sigma and a threshold D, we would like to find all short substrings of S of length P that repeat with at most D differences, i.e., insertions, deletions, and mismatches. We give a careful theoretical characterization of the set of seeds (i.e., some maximal exact repeats) required by the algorithm, and prove a sublinear bound on their expected numbers. Using this result, we present a sub-quadratic algorithm for finding all short (i.e., of length O(log N)) approximate repeats. The running time of our algorithm is O(DN(3pow(epsilon)-1)log N), where epsilon = D/P and pow(epsilon) is an increasing, concave function that is 0 when epsilon = 0 and about 0.9 for DNA and protein sequences. PMID- 11472988 TI - SCOPE: a probabilistic model for scoring tandem mass spectra against a peptide database. AB - Proteomics, or the direct analysis of the expressed protein components of a cell, is critical to our understanding of cellular biological processes in normal and diseased tissue. A key requirement for its success is the ability to identify proteins in complex mixtures. Recent technological advances in tandem mass spectrometry has made it the method of choice for high-throughput identification of proteins. Unfortunately, the software for unambiguously identifying peptide sequences has not kept pace with the recent hardware improvements in mass spectrometry instruments. Critical for reliable high-throughput protein identification, scoring functions evaluate the quality of a match between experimental spectra and a database peptide. Current scoring function technology relies heavily on ad-hoc parameterization and manual curation by experienced mass spectrometrists. In this work, we propose a two-stage stochastic model for the observed MS/MS spectrum, given a peptide. Our model explicitly incorporates fragment ion probabilities, noisy spectra, and instrument measurement error. We describe how to compute this probability based score efficiently, using a dynamic programming technique. A prototype implementation demonstrates the effectiveness of the model. PMID- 11472989 TI - Fast optimal leaf ordering for hierarchical clustering. AB - We present the first practical algorithm for the optimal linear leaf ordering of trees that are generated by hierarchical clustering. Hierarchical clustering has been extensively used to analyze gene expression data, and we show how optimal leaf ordering can reveal biological structure that is not observed with an existing heuristic ordering method. For a tree with n leaves, there are 2(n-1) linear orderings consistent with the structure of the tree. Our optimal leaf ordering algorithm runs in time O(n(4)), and we present further improvements that make the running time of our algorithm practical. PMID- 11472990 TI - Separating real motifs from their artifacts. AB - The typical output of many computational methods to identify binding sites is a long list of motifs containing some real motifs (those most likely to correspond to the actual binding sites) along with a large number of random variations of these. We present a statistical method to separate real motifs from their artifacts. This produces a short list of high quality motifs that is sufficient to explain the over-representation of all motifs in the given sequences. Using synthetic data sets, we show that the output of our method is very accurate. On various sets of upstream sequences in S. cerevisiae, our program identifies several known binding sites, as well as a number of significant novel motifs. PMID- 11472991 TI - Probe selection algorithms with applications in the analysis of microbial communities. AB - We propose two efficient heuristics for minimizing the number of oligonucleotide probes needed for analyzing populations of ribosomal RNA gene (rDNA) clones by hybridization experiments on DNA microarrays. Such analyses have applications in the study of microbial communities. Unlike in the classical SBH (sequencing by hybridization) procedure, where multiple probes are on a DNA chip, in our applications we perform a series of experiments, each one consisting of applying a single probe to a DNA microarray containing a large sample of rDNA sequences from the studied population. The overall cost of the analysis is thus roughly proportional to the number of experiments, underscoring the need for minimizing the number of probes. Our algorithms are based on two well-known optimization techniques, i.e. simulated annealing and Lagrangian relaxation, and our preliminary tests demonstrate that both algorithms are able to find satisfactory probe sets for real rDNA data. PMID- 11472992 TI - Visualizing associations between genome sequences and gene expression data using genome-mean expression profiles. AB - The combination of genome-wide expression patterns and full genome sequences offers a great opportunity to further our understanding of the mechanisms and logic of transcriptional regulation. Many methods have been described that identify sequence motifs enriched in transcription control regions of genes that share similar gene expression patterns. Here we present an alternative approach that evaluates the transcriptional information contained by specific sequence motifs by computing for each motif the mean expression profile of all genes that contain the motif in their transcription control regions. These genome-mean expression profiles (GMEP's) are valuable for visualizing the relationship between genome sequences and gene expression data, and for characterizing the transcriptional importance of specific sequence motifs. Analysis of GMEP's calculated from a dataset of 519 whole-genome microarray experiments in Saccharomyces cerevisiae show a significant correlation between GMEP's of motifs that are reverse complements, a result that supports the relationship between GMEP's and transcriptional regulation. Hierarchical clustering of GMEP's identifies clusters of motifs that correspond to binding sites of well characterized transcription factors. The GMEP's of these clustered motifs have patterns of variation across conditions that reflect the known activities of these transcription factors. Software that computed GMEP's from sequence and gene expression data is available under the terms of the Gnu Public License from http://rana.lbl.gov/. PMID- 11472993 TI - Gene recognition based on DAG shortest paths. AB - We describe DAGGER, an ab initio gene recognition program which combines the output of high dimensional signal sensors in an intuitive gene model based on directed acyclic graphs. In the first stage, candidate start, donor, acceptor, and stop sites are scored using the SNoW learning architecture. These sites are then used to generate a directed acyclic graph in which each source-sink path represents a possible gene structure. Training sequences are used to optimize an edge weighting function so that the shortest source-sink path maximizes exon level prediction accuracy. Experimental evaluation of prediction accuracy on two benchmark data sets demonstrates that DAGGERis competitive with ab initio gene finding programs based on Hidden Markov Models. PMID- 11472994 TI - Using mixtures of common ancestors for estimating the probabilities of discrete events in biological sequences. AB - Accurately estimating probabilities from observations is important for probabilistic-based approaches to problems in computational biology. In this paper we present a biologically-motivated method for estimating probability distributions over discrete alphabets from observations using a mixture model of common ancestors. The method is an extension of substitution matrix-based probability estimation methods. In contrast to previous such methods, our method has a simple Bayesian interpretation and has the advantage over Dirichlet mixtures that it is both effective and simple to compute for large alphabets. The method is applied to estimate amino acid probabilities based on observed counts in an alignment and is shown to perform comparably to previous methods. The method is also applied to estimate probability distributions over protein families and improves protein classification accuracy. PMID- 11472995 TI - GENIES: a natural-language processing system for the extraction of molecular pathways from journal articles. AB - Systems that extract structured information from natural language passages have been highly successful in specialized domains. The time is opportune for developing analogous applications for molecular biology and genomics. We present a system, GENIES, that extracts and structures information about cellular pathways from the biological literature in accordance with a knowledge model that we developed earlier. We implemented GENIES by modifying an existing medical natural language processing system, MedLEE, and performed a preliminary evaluation study. Our results demonstrate the value of the underlying techniques for the purpose of acquiring valuable knowledge from biological journals. PMID- 11472996 TI - An insight into domain combinations. AB - Domains are the building blocks of all globular proteins, and are units of compact three-dimensional structure as well as evolutionary units. There is a limited repertoire of domain families, so that these domain families are duplicated and combined in different ways to form the set of proteins in a genome. Proteins are gene products. The processes that produce new genes are duplication and recombination as well as gene fusion and fission. We attempt to gain an overview of these processes by studying the structural domains in the proteins of seven genomes from the three kingdoms of life: Eubacteria, Archaea and Eukaryota. We use here the domain and superfamily definitions in Structural Classification of Proteins Database (SCOP) in order to map pairs of adjacent domains in genome sequences in terms of their superfamily combinations. We find 624 out of the 764 superfamilies in SCOP in these genomes, and the 624 families occur in 585 pairwise combinations. Most families are observed in combination with one or two other families, while a few families are very versatile in their combinatorial behaviour. This type of pattern can be described by a scale-free network. Finally, we study domain repeats and we compare the set of the domain combinations in the genomes to those in PDB, and discuss the implications for structural genomics. PMID- 11472997 TI - Promoter prediction in the human genome. AB - Computational prediction of eukaryotic poIII promoters has been one of the most elusive problems despite considerable effort devoted to the study. Researchers have looked for various types of signals around the transcriptional start site (TSS), viz. oligo-nucleotide statistics, potential binding sites for core factors, clusters of binding sites, proximity to CpG islands etc. The proximity of CpG islands to gene starts is now a well established fact, although until recently, it was based on very little genomic data. In this work we explore the possibility of enhancing the promoter prediction accuracy by combining CpG island information with a few other, biologically motivated, seemingly independent signals, that cover most of the known knowledge. We benchmarked the method on a much larger genomic datasets compared to previous studies. We were able to improve slightly upon current prediction accuracy. Furthermore, we observe that CpG islands are the most dominant signals and the other signals do not improve the prediction. This suggests that the computational prediction of promoters for genes with no associated CpG-island (typically having tissue-specific expression) looking only at the immediate neighborhood of the TSS may not even be possible. We suggest some biological experiments and studies to better understand the biology of transcription. PMID- 11472998 TI - Disambiguating proteins, genes, and RNA in text: a machine learning approach. AB - We present an automated system for assigning protein, gene, or mRNA class labels to biological terms in free text. Three machine learning algorithms and several extended ways for defining contextual features for disambiguation are examined, and a fully unsupervised manner for obtaining training examples is proposed. We train and evaluate our system over a collection of 9 million words of molecular biology journal articles, obtaining accuracy rates up to 85%. PMID- 11472999 TI - Identifying splits with clear separation: a new class discovery method for gene expression data. AB - We present a new class discovery method for microarray gene expression data. Based on a collection of gene expression profiles from different tissue samples, the method searches for binary class distinctions in the set of samples that show clear separation in the expression levels of specific subsets of genes. Several mutually independent class distinctions may be found, which is difficult to obtain from most commonly used clustering algorithms. Each class distinction can be biologically interpreted in terms of its supporting genes. The mathematical characterization of the favored class distinctions is based on statistical concepts. By analyzing three data sets from cancer gene expression studies, we demonstrate that our method is able to detect biologically relevant structures, for example cancer subtypes, in an unsupervised fashion. PMID- 11473000 TI - GEST: a gene expression search tool based on a novel Bayesian similarity metric. AB - Gene expression array technology has made possible the assay of expression levels of tens of thousands of genes at a time; large databases of such measurements are currently under construction. One important use of such databases is the ability to search for experiments that have similar gene expression levels as a query, potentially identifying previously unsuspected relationships among cellular states. Such searches depend crucially on the metric used to assess the similarity between pairs of experiments. The complex joint distribution of gene expression levels, particularly their correlational structure and non-normality, make simple similarity metrics such as Euclidean distance or correlational similarity scores suboptimal for use in this application. We present a similarity metric for gene expression array experiments that takes into account the complex joint distribution of expression values. We provide a computationally tractable approximation to this measure, and have implemented a database search tool based on it. We discuss implementation issues and efficiency, and we compare our new metric to other standard metrics. PMID- 11473001 TI - Probabilistic divergence measures for detecting interspecies recombination. AB - This paper proposes a graphical method for detecting interspecies recombination in multiple alignments of DNA sequences. A fixed-size window is moved along a given DNA sequence alignment. For every position, the marginal posterior probability over tree topologies is determined by means of a Markov chain Monte Carlo simulation. Two probabilistic divergence measures are plotted along the alignment, and are used to identify recombinant regions. The method is compared with established detection methods on a set of synthetic benchmark sequences and two real-world DNA sequence alignments. PMID- 11473002 TI - Design of a compartmentalized shotgun assembler for the human genome. AB - Two different strategies for determining the human genome are currently being pursued: one is the "clone-by-clone" approach, employed by the publicly funded project, and the other is the "whole genome shotgun assembler" approach, favored by researchers at Celera Genomics. An interim strategy employed at Celera, called compartmentalized shotgun assembly, makes use of preliminary data produced by both approaches. In this paper we describe the design, implementation and operation of the "compartmentalized shotgun assembler". PMID- 11473003 TI - Integrating genomic homology into gene structure prediction. AB - TWINSCAN is a new gene-structure prediction system that directly extends the probability model of GENSCAN, allowing it to exploit homology between two related genomes. Separate probability models are used for conservation in exons, introns, splice sites, and UTRs, reflecting the differences among their patterns of evolutionary conservation. TWINSCAN is specifically designed for the analysis of high-throughput genomic sequences containing an unknown number of genes. In experiments on high-throughput mouse sequences, using homologous sequences from the human genome, TWINSCAN shows notable improvement over GENSCAN in exon sensitivity and specificity and dramatic improvement in exact gene sensitivity and specificity. This improvement can be attributed entirely to modeling the patterns of evolutionary conservation in genomic sequence. PMID- 11473004 TI - Generating protein interaction maps from incomplete data: application to fold assignment. AB - MOTIVATION: We present a framework to generate comprehensive overviews of protein protein interactions. In the post-genomic view of cellular function, each biological entity is seen in the context of a complex network of interactions. Accordingly, we model functional space by representing protein-protein interaction data as undirected graphs. We suggest a general approach to generate interaction maps of cellular networks in the presence of huge amounts of fragmented and incomplete data, and to derive representations of large networks which hide clutter while keeping the essential architecture of the interaction space. This is achieved by contracting the graphs according to domain-specific hierarchical classifications. The key concept here is the notion of induced interaction, which allows the integration, comparison and analysis of interaction data from different sources and different organisms at a given level of abstraction. RESULTS: We apply this approach to compute the overlap between the DIP compendium of interaction data and a dataset of yeast two-hybrid experiments. The architecture of this network is scale-free, as frequently seen in biological networks, and this property persists through many levels of abstraction. Connections in the network can be projected downwards from higher levels of abstraction down to the level of individual proteins. As an example, we describe an algorithm for fold assignment by network context. This method currently predicts protein folds at 30% accuracy without any requirement of detectable sequence similarity of the query protein to a protein of known structure. We used this algorithm to compile a list of structural assignments for previously unassigned genes from yeast. Finally we discuss ways forward to use interaction networks for the prediction of novel protein-protein interactions. AVAILABILITY: http://www.ebi.ac.uk/~lappe/FoldPred/. PMID- 11473005 TI - Feature selection for DNA methylation based cancer classification. AB - Molecular portraits, such as mRNA expression or DNA methylation patterns, have been shown to be strongly correlated with phenotypical parameters. These molecular patterns can be revealed routinely on a genomic scale. However, class prediction based on these patterns is an under-determined problem, due to the extreme high dimensionality of the data compared to the usually small number of available samples. This makes a reduction of the data dimensionality necessary. Here we demonstrate how phenotypic classes can be predicted by combining feature selection and discriminant analysis. By comparing several feature selection methods we show that the right dimension reduction strategy is of crucial importance for the classification performance. The techniques are demonstrated by methylation pattern based discrimination between acute lymphoblastic leukemia and acute myeloid leukemia. PMID- 11473006 TI - New approaches for reconstructing phylogenies from gene order data. AB - We report on new techniques we have developed for reconstructing phylogenies on whole genomes. Our mathematical techniques include new polynomial-time methods for bounding the inversion length of a candidate tree and new polynomial-time methods for estimating genomic distances which greatly improve the accuracy of neighbor-joining analyses. We demonstrate the power of these techniques through an extensive performance study based on simulating genome evolution under a wide range of model conditions. Combining these new tools with standard approaches (fast reconstruction with neighbor-joining, exploration of all possible refinements of strict consensus trees, etc.) has allowed us to analyze datasets that were previously considered computationally impractical. In particular, we have conducted a complete phylogenetic analysis of a subset of the Campanulaceae family, confirming various conjectures about the relationships among members of the subset and about the principal mechanism of evolution for their chloroplast genome. We give representative results of the extensive experimentation we conducted on both real and simulated datasets in order to validate and characterize our approaches. We find that our techniques provide very accurate reconstructions of the true tree topology even when the data are generated by processes that include a significant fraction of transpositions and when the data are close to saturation. PMID- 11473007 TI - Prediction of the coupling specificity of G protein coupled receptors to their G proteins. AB - G protein coupled receptors (GPCRs) are found in great numbers in most eukaryotic genomes. They are responsible for sensing a staggering variety of structurally diverse ligands, with their activation resulting in the initiation of a variety of cellular signalling cascades. The physiological response that is observed following receptor activation is governed by the guanine nucleotide-binding proteins (G proteins) to which a particular receptor chooses to couple. Previous investigations have demonstrated that the specificity of the receptor-G protein interaction is governed by the intracellular domains of the receptor. Despite many studies it has proven very difficult to predict de novo, from the receptor sequence alone, the G proteins to which a GPCR is most likely to couple. We have used a data-mining approach, combining pattern discovery with membrane topology prediction, to find patterns of amino acid residues in the intracellular domains of GPCR sequences that are specific for coupling to a particular functional class of G proteins. A prediction system was then built, being based upon these discovered patterns. We can report this approach was successful in the prediction of G protein coupling specificity of unknown sequences. Such predictions should be of great use in providing in silico characterisation of newly cloned receptor sequences and for improving the annotation of GPCRs stored in protein sequence databases. AVAILABILITY: http://www.ebi.ac.uk/~croning/coupling.html. PMID- 11473008 TI - Non-symmetric score matrices and the detection of homologous transmembrane proteins. AB - Given a transmembrane protein, we wish to find related ones by a database search. Due to the strongly hydrophobic amino acid composition of transmembrane domains, suboptimal results are obtained when general-purpose scoring matrices such as BLOSUM are used. Recently, a transmembrane-specific score matrix called PHAT was shown to perform much better than BLOSUM. In this article, we derive a transmembrane score matrix family, called SLIM, which has several distinguishing features. In contrast to currently used matrices, SLIM is non-symmetric. The asymmetry arises because different background compositions are assumed for the transmembrane query and the unknown database sequences. We describe the mathematical model behind SLIM in detail and show that SLIM outperforms PHAT both on simulated data and in a realistic setting. Since non-symmetric score matrices are a new concept in database search methods, we discuss some important theoretical and practical issues. PMID- 11473009 TI - Designing fast converging phylogenetic methods. AB - Absolute fast converging phylogenetic reconstruction methods are provably guaranteed to recover the true tree with high probability from sequences that grow only polynomially in the number of leaves, once the edge lengths are bounded arbitrarily from above and below. Only a few methods have been determined to be absolute fast converging; these have all been developed in just the last few years, and most are polynomial time. In this paper, we compare pre-existing fast converging methods as well as some new polynomial time methods that we have developed. Our study, based upon simulating evolution under a wide range of model conditions, establishes that our new methods outperform both neighbor joining and the previous fast converging methods, returning very accurate large trees, when these other methods do poorly. PMID- 11473010 TI - Joint modeling of DNA sequence and physical properties to improve eukaryotic promoter recognition. AB - We present an approach to integrate physical properties of DNA, such as DNA bendability or GC content, into our probabilistic promoter recognition system McPROMOTER. In the new model, a promoter is represented as a sequence of consecutive segments represented by joint likelihoods for DNA sequence and profiles of physical properties. Sequence likelihoods are modeled with interpolated Markov chains, physical properties with Gaussian distributions. The background uses two joint sequence/profile models for coding and non-coding sequences, each consisting of a mixture of a sense and an anti-sense submodel. On a large Drosophila test set, we achieved a reduction of about 30% of false positives when compared with a model solely based on sequence likelihoods. PMID- 11473011 TI - An algorithm for finding signals of unknown length in DNA sequences. AB - Pattern discovery in unaligned DNA sequences is a challenging problem in both computer science and molecular biology. Several different methods and techniques have been proposed so far, but in most of the cases signals in DNA sequences are very complicated and avoid detection. Exact exhaustive methods can solve the problem only for short signals with a limited number of mutations. In this work, we extend exhaustive enumeration also to longer patterns. More in detail, the basic version of algorithm presented in this paper, given as input a set of sequences and an error ratio epsilon < 1, finds all patterns that occur in at least q sequences of the set with at most epsilonm mutations, where m is the length of the pattern. The only restriction is imposed on the location of mutations along the signal. That is, a valid occurrence of a pattern can present at most [epsiloni] mismatches in the first i nucleotides, and so on. However, we show how the algorithm can be used also when no assumption can be made on the position of mutations. In this case, it is also possible to have an estimate of the probability of finding a signal according to the signal length, the error ratio, and the input parameters. Finally, we discuss some significance measures that can be used to sort the patterns output by the algorithm. PMID- 11473012 TI - Inferring subnetworks from perturbed expression profiles. AB - Genome-wide expression profiles of genetic mutants provide a wide variety of measurements of cellular responses to perturbations. Typical analysis of such data identifies genes affected by perturbation and uses clustering to group genes of similar function. In this paper we discover a finer structure of interactions between genes, such as causality, mediation, activation, and inhibition by using a Bayesian network framework. We extend this framework to correctly handle perturbations, and to identify significant subnetworks of interacting genes. We apply this method to expression data of S. cerevisiae mutants and uncover a variety of structured metabolic, signaling and regulatory pathways. PMID- 11473013 TI - Fragment assembly with double-barreled data. AB - For the last twenty years fragment assembly was dominated by the "overlap - layout - consensus" algorithms that are used in all currently available assembly tools. However, the limits of these algorithms are being tested in the era of genomic sequencing and it is not clear whether they are the best choice for large scale assemblies. Although the "overlap - layout - consensus" approach proved to be useful in assembling clones, it faces difficulties in genomic assemblies: the existing algorithms make assembly errors even in bacterial genomes. We abandoned the "overlap - layout - consensus" approach in favour of a new Eulerian Superpath approach that outperforms the existing algorithms for genomic fragment assembly (Pevzner et al. 2001 InProceedings of the Fifth Annual International Conference on Computational Molecular Biology (RECOMB-01), 256-26). In this paper we describe our new EULER-DB algorithm that, similarly to the Celera assembler takes advantage of clone-end sequencing by using the double-barreled data. However, in contrast to the Celera assembler, EULER-DB does not mask repeats but uses them instead as a powerful tool for contig ordering. We also describe a new approach for the Copy Number Problem: "How many times a given repeat is present in the genome?". For long nearly-perfect repeats this question is notoriously difficult and some copies of such repeats may be "lost" in genomic assemblies. We describe our EULER-CN algorithm for the Copy Number Problem that proved to be successful in difficult sequencing projects. PMID- 11473014 TI - Improved prediction of the number of residue contacts in proteins by recurrent neural networks. AB - Knowing the number of residue contacts in a protein is crucial for deriving constraints useful in modeling protein folding, protein structure, and/or scoring remote homology searches. Here we use an ensemble of bi-directional recurrent neural network architectures and evolutionary information to improve the state-of the-art in contact prediction using a large corpus of curated data. The ensemble is used to discriminate between two different states of residue contacts, characterized by a contact number higher or lower than the average value of the residue distribution. The ensemble achieves performances ranging from 70.1% to 73.1% depending on the radius adopted to discriminate contacts (6Ato 12A). These performances represent gains of 15% to 20% over the base line statistical predictors always assigning an aminoacid to the most numerous state, 3% to 7% better than any previous method. Combination of different radius predictors further improves the performance. SERVER: http://promoter.ics.uci.edu/BRNN-PRED/. PMID- 11473015 TI - Rich probabilistic models for gene expression. AB - Clustering is commonly used for analyzing gene expression data. Despite their successes, clustering methods suffer from a number of limitations. First, these methods reveal similarities that exist over all of the measurements, while obscuring relationships that exist over only a subset of the data. Second, clustering methods cannot readily incorporate additional types of information, such as clinical data or known attributes of genes. To circumvent these shortcomings, we propose the use of a single coherent probabilistic model, that encompasses much of the rich structure in the genomic expression data, while incorporating additional information such as experiment type, putative binding sites, or functional information. We show how this model can be learned from the data, allowing us to discover patterns in the data and dependencies between the gene expression patterns and additional attributes. The learned model reveals context-specific relationships, that exist only over a subset of the experiments in the dataset. We demonstrate the power of our approach on synthetic data and on two real-world gene expression data sets for yeast. For example, we demonstrate a novel functionality that falls naturally out of our framework: predicting the "cluster" of the array resulting from a gene mutation based only on the gene's expression pattern in the context of other mutations. PMID- 11473016 TI - Designing better phages. AB - We propose a method to engineer the genome of bacteriophages to increase their effectiveness as antibacterial agents. Specifically, we exploit the redundancy of the triplet code to design genomes that avoid restriction sites while producing the same proteins as wild-type phages. We give an efficient algorithm to minimize the number of restriction sites against sets of cutter sequences, and demonstrate that that phage genomes can be significantly protected against surprisingly large sets of enzymes with no loss of function. Finally, we develop a model to explain why evolution has failed to eliminate many possible restriction sites despite selective pressure, thus motivating the need for genome-level sequence engineering. PMID- 11473017 TI - Probabilistic approaches to the use of higher order clone relationships in physical map assembly. AB - Physical map assembly is the inference of genome structure from experimental data. Map assembly depends on the integration of diverse data including sequence tagged site (STS) marker content, clone sizing, and restriction digest fingerprints (RDF). As experimentally measured data, these are uncertain and error prone. Physical map assembly from error free data is straightforward and can be accomplished in linear time in the number of clones, but the assembly of an optimal map from error prone data is an NP-hard problem. We present an alternative approach to physical map assembly that is based on a probabilistic view of the data and seeks to identify those features of the map that can be reliably inferred from the available data. With this approach, we achieve a number of goals. These include the use of multiple data sources, appropriate representation of uncertainties in the underlying data, the use of clone length information in fingerprint map assembly, and the use of higher order information in map assembly. By higher order information, we mean relationships that are not expressible in terms of neighbouring clone relationships. These include triplet and multiple clone overlaps, the uniqueness of STS position, and fingerprint marker locations. In a probabilistic view of physical mapping, we assert that all of the many possible map assemblies are equally likely a priori. Given experimental data, we can only state which assemblies are more likely than others given the experimental observations. Parameters of interest are then derived as likelihood weighted averages over map assemblies. Ideally these averages should be sums or integrals over all possible map assemblies, but computationally this is not feasible for real-world map assembly problems. Instead, sampling is used to asymptotically approach the desired parameters. Software implementing our probabilistic approach to mapping has been written. Assembly of mixed RDF and STS maps containing up to 60 clones can be accomplished on a desktop PC with run times under an hour. AVAILABILITY: http://stl.wustl.edu/software/gibbsmap/. PMID- 11473018 TI - Computational expansion of genetic networks. AB - We present a new methodology for computational analysis of gene and protein networks. The aim is to generate new educated hypotheses on gene functions and on the logic of the biological network circuitry, based on gene expression profiles. The framework supports the incorporation of biologically motivated network constraints and rules to improve specificity. Since current data is insufficient for de-novo reconstruction, the method receives as input a known pathway core and suggests likely expansions to it. Network modeling is combinatorial, yet data can be probabilistic. At the heart of the approach are a fitness function which estimates the quality of suggested network expansions given the core and the data, and a specificity measure of the expansions. The approach has been implemented in an interactive software tool called GENESYS. We report encouraging results in preliminary analysis of yeast ergosterol pathway based on transcription profiles. In particular, the analysis suggests a novel ergosterol transcription factor. PMID- 11473019 TI - Separation of samples into their constituents using gene expression data. AB - Gene expression measurements are a powerful tool in molecular biology, but when applied to heterogeneous samples containing more than one cellular type the results are difficult to interpret. We present here a new approach to this problem allowing to deduce the gene expression profile of the various cellular types contained in a set of samples directly from the measurements taken on the whole sample. PMID- 11473020 TI - 0j.py: a software tool for low complexity proteins and protein domains. AB - Low complexity proteins and protein domains have sequences which appear highly non-random. Over the years, these sequences have been routinely filtered out during sequence similarity searches because interest has been focused on globular proteins, and inclusion of these domains can severely skew search results. However, early work on these proteins and more recent studies of the related area of repeated protein sequences suggests that low complexity protein domains have function and therefore are in need of further investigation. 0j.py is a new tool for demarcating low complexity protein domains more accurately than has been possible to date. The paper describes 0j.py and its use in revealing proteins with repeated and poly-amino-acid peptides. Statistical methods are then employed to to examine the distribution of these proteins across species, while keyword clustering is used to suggest roles performed by proteins through the use of low complexity domains. PMID- 11473021 TI - Protein-protein interaction map inference using interacting domain profile pairs. AB - A number of predictive methods have been designed to predict protein interaction from sequence or expression data. On the experimental front, however, high throughput proteomics technologies are starting to yield large volumes of protein protein interaction data. High-quality experimental protein interaction maps constitute the natural dataset upon which to build interaction predictions. Thus the motivation to develop the first interaction-based protein interaction map prediction algorithm. A technique to predict protein-protein interaction maps across organisms is introduced, the 'interaction-domain pair profile' method. The method uses a high-quality protein interaction map with interaction domain information as input to predict an interaction map in another organism. It combines sequence similarity searches with clustering based on interaction patterns and interaction domain information. We apply this approach to the prediction of an interaction map of Escherichia coli from the recently published interaction map of the human gastric pathogen Helicobacter pylori. Results are compared with predictions of a second inference method based only on full-length protein sequence similarity - the "naive" method. The domain-based method is shown to i) eliminate a significant amount of false-positives of the naive method that are the consequences of multi-domain proteins; ii) increase the sensitivity compared to the naive method by identifying new potential interactions. AVAILABILITY: Contact the authors. PMID- 11473022 TI - CLIFF: clustering of high-dimensional microarray data via iterative feature filtering using normalized cuts. AB - We present CLIFF, an algorithm for clustering biological samples using gene expression microarray data. This clustering problem is difficult for several reasons, in particular the sparsity of the data, the high dimensionality of the feature (gene) space, and the fact that many features are irrelevant or redundant. Our algorithm iterates between two computational processes, feature filtering and clustering. Given a reference partition that approximates the correct clustering of the samples, our feature filtering procedure ranks the features according to their intrinsic discriminability, relevance to the reference partition, and irredundancy to other relevant features, and uses this ranking to select the features to be used in the following round of clustering. Our clustering algorithm, which is based on the concept of a normalized cut, clusters the samples into a new reference partition on the basis of the selected features. On a well-studied problem involving 72 leukemia samples and 7130 genes, we demonstrate that CLIFF outperforms standard clustering approaches that do not consider the feature selection issue, and produces a result that is very close to the original expert labeling of the sample set. PMID- 11473023 TI - Molecular classification of multiple tumor types. AB - Using gene expression data to classify tumor types is a very promising tool in cancer diagnosis. Previous works show several pairs of tumor types can be successfully distinguished by their gene expression patterns (Golub et al. 1999, Ben-Dor et al. 2000, Alizadeh et al. 2000). However, the simultaneous classification across a heterogeneous set of tumor types has not been well studied yet. We obtained 190 samples from 14 tumor classes and generated a combined expression dataset containing 16063 genes for each of those samples. We performed multi-class classification by combining the outputs of binary classifiers. Three binary classifiers (k-nearest neighbors, weighted voting, and support vector machines) were applied in conjunction with three combination scenarios (one-vs-all, all-pairs, hierarchical partitioning). We achieved the best cross validation error rate of 18.75% and the best test error rate of 21.74% by using the one-vs-all support vector machine algorithm. The results demonstrate the feasibility of performing clinically useful classification from samples of multiple tumor types. PMID- 11473024 TI - Centralization: a new method for the normalization of gene expression data. AB - Microarrays measure values that are approximately proportional to the numbers of copies of different mRNA molecules in samples. Due to technical difficulties, the constant of proportionality between the measured intensities and the numbers of mRNA copies per cell is unknown and may vary for different arrays. Usually, the data are normalized (i.e., array-wise multiplied by appropriate factors) in order to compensate for this effect and to enable informative comparisons between different experiments. Centralization is a new two-step method for the computation of such normalization factors that is both biologically better motivated and more robust than standard approaches. First, for each pair of arrays the quotient of the constants of proportionality is estimated. Second, from the resulting matrix of pairwise quotients an optimally consistent scaling of the samples is computed. PMID- 11473025 TI - Glucose induces beta-cell apoptosis via upregulation of the Fas receptor in human islets. AB - In autoimmune type 1 diabetes, Fas-to-Fas-ligand (FasL) interaction may represent one of the essential pro-apoptotic pathways leading to a loss of pancreatic beta cells. In the advanced stages of type 2 diabetes, a decline in beta-cell mass is also observed, but its mechanism is not known. Human islets normally express FasL but not the Fas receptor. We observed upregulation of Fas in beta-cells of type 2 diabetic patients relative to nondiabetic control subjects. In vitro exposure of islets from nondiabetic organ donors to high glucose levels induced Fas expression, caspase-8 and -3 activation, and beta-cell apoptosis. The effect of glucose was blocked by an antagonistic anti-Fas antibody, indicating that glucose induced apoptosis is due to interaction between the constitutively expressed FasL and the upregulated Fas. These results support a new role for glucose in regulating Fas expression in human beta-cells. Upregulation of the Fas receptor by elevated glucose levels may contribute to beta-cell destruction by the constitutively expressed FasL independent of an autoimmune reaction, thus providing a link between type 1 and type 2 diabetes. PMID- 11473026 TI - Insulin production by human embryonic stem cells. AB - Type 1 diabetes generally results from autoimmune destruction of pancreatic islet beta-cells, with consequent absolute insulin deficiency and complete dependence on exogenous insulin treatment. The relative paucity of donations for pancreas or islet allograft transplantation has prompted the search for alternative sources for beta-cell replacement therapy. In the current study, we used pluripotent undifferentiated human embryonic stem (hES) cells as a model system for lineage specific differentiation. Using hES cells in both adherent and suspension culture conditions, we observed spontaneous in vitro differentiation that included the generation of cells with characteristics of insulin-producing beta-cells. Immunohistochemical staining for insulin was observed in a surprisingly high percentage of cells. Secretion of insulin into the medium was observed in a differentiation-dependent manner and was associated with the appearance of other beta-cell markers. These findings validate the hES cell model system as a potential basis for enrichment of human beta-cells or their precursors, as a possible future source for cell replacement therapy in diabetes. PMID- 11473027 TI - Complete protection of islets against allorejection and autoimmunity by a simple barium-alginate membrane. AB - We describe a new technique for microencapsulation with high-mannuronic acid (high-M) alginate crosslinked with BaCl(2) without a traditional permselective component, which allows the production of biocompatible capsules that allow prolonged survival of syngeneic and allogeneic transplanted islets in diabetic BALB/c and NOD mice for >350 days. The normalization of the glycemia in the transplanted mice was associated with normal glucose profiles in response to intravenous glucose tolerance tests. After explantation of the capsules, all mice became hyperglycemic, demonstrating the efficacy of the encapsulated islets. The retrieved capsules were free of cellular overgrowth and islets responded to glucose stimulation with a 5- to 10-fold increase of insulin secretion. Transfer of splenocytes isolated from transplanted NOD mice to NOD/SCID mice adoptively transferred diabetes, indicating that NOD recipients maintained islet-specific autoimmunity. In conclusion, we have developed a simple technique for microencapsulation that prolongs islet survival without immunosuppression, providing complete protection against allorejection and the recurrence of autoimmune diabetes. PMID- 11473028 TI - Proteins linked to a protein transduction domain efficiently transduce pancreatic islets. AB - The resounding success of a new immunosuppressive regimen known as the Edmonton protocol demonstrates that islet cell transplantation is becoming a therapeutic reality for diabetes. However, under the Edmonton protocol, a single donor does not provide enough islets to attain the insulin independence of a transplant recipient. This limitation is mainly caused by islet apoptosis triggered during isolation. In this study, we describe a highly efficient system of transiently transferring anti-apoptotic proteins into pancreatic islets, thus opening an exciting new therapeutic opportunity to improve the viability of transplantable islets. We fused beta-galactosidase to the 11-amino acid residues that constitute the protein transduction domain (PTD) of the HIV/TAT protein and transduced pancreatic islets ex vivo with this fusion protein in a dose-dependent manner with >80% efficiency. We observed that transduction of the anti-apoptotic proteins Bcl-X(L) and PEA-15 fused to TAT/PTD prevented apoptosis induced by tumor necrosis factor-alpha in a pancreatic beta-cell line, indicating that TAT/PTD anti-apoptotic proteins retained their biological activity. Finally, we demonstrated that TAT-fusion proteins did not affect the insulin secretion capability of islets, as determined by glucose static incubation and by reversion of hyperglycemia in diabetic immunodeficient mice. PMID- 11473029 TI - A preprandial rise in plasma ghrelin levels suggests a role in meal initiation in humans. AB - The recently discovered orexigenic peptide ghrelin is produced primarily by the stomach and circulates in blood at levels that increase during prolonged fasting in rats. When administered to rodents at supraphysiological doses, ghrelin activates hypothalamic neuropeptide Y/agouti gene-related protein neurons and increases food intake and body weight. These findings suggest that ghrelin may participate in meal initiation. As a first step to investigate this hypothesis, we sought to determine whether circulating ghrelin levels are elevated before the consumption of individual meals in humans. Ghrelin, insulin, and leptin were measured by radioimmunoassay in plasma samples drawn 38 times throughout a 24-h period in 10 healthy subjects provided meals on a fixed schedule. Plasma ghrelin levels increased nearly twofold immediately before each meal and fell to trough levels within 1 h after eating, a pattern reciprocal to that of insulin. Intermeal ghrelin levels displayed a diurnal rhythm that was exactly in phase with that of leptin, with both hormones rising throughout the day to a zenith at 0100, then falling overnight to a nadir at 0900. Ghrelin levels sampled during the troughs before and after breakfast correlated strongly with 24-h integrated area under the curve values (r = 0.873 and 0.954, respectively), suggesting that these convenient, single measurements might serve as surrogates for 24-h profiles to estimate overall ghrelin levels. Circulating ghrelin also correlated positively with age (r = 0.701). The clear preprandial rise and postprandial fall in plasma ghrelin levels support the hypothesis that ghrelin plays a physiological role in meal initiation in humans. PMID- 11473030 TI - Glucose competence of the hepatoportal vein sensor requires the presence of an activated glucagon-like peptide-1 receptor. AB - Activation of the hepatoportal glucose sensors by portal glucose infusion leads to increased glucose clearance and induction of hypoglycemia. Here, we investigated whether glucagon-like peptide-1 (GLP-1) could modulate the activity of these sensors. Mice were therefore infused with saline (S-mice) or glucose (P mice) through the portal vein at a rate of 25 mg/kg. min. In P-mice, glucose clearance increased to 67.5 +/- 3.7 mg/kg. min as compared with 24.1 +/- 1.5 mg/kg. min in S-mice, and glycemia decreased from 5.0 +/- 0.1 to 3.3 +/- 0.1 mmol/l at the end of the 3-h infusion period. Coinfusion of GLP-1 with glucose into the portal vein at a rate of 5 pmol/kg. min (P-GLP-1 mice) did not increase the glucose clearance rate (57.4 +/- 5.0 ml/kg. min) and hypoglycemia (3.8 +/- 0.1 mmol/l) observed in P-mice. In contrast, coinfusion of glucose and the GLP-1 receptor antagonist exendin-(9-39) into the portal vein at a rate of 0.5 pmol/kg. min (P-Ex mice) reduced glucose clearance to 36.1 +/- 2.6 ml/kg. min and transiently increased glycemia to 9.2 +/- 0.3 mmol/l at 60 min of infusion before it returned to the fasting level (5.6 +/- 0.3 mmol/l) at 3 h. When glucose and exendin-(9-39) were infused through the portal and femoral veins, respectively, glucose clearance increased to 70.0 +/- 4.6 ml/kg. min and glycemia decreased to 3.1 +/- 0.1 mmol/l, indicating that exendin-(9-39) has an effect only when infused into the portal vein. Finally, portal vein infusion of glucose in GLP-1 receptor(-/-) mice failed to increase the glucose clearance rate (26.7 +/- 2.9 ml/kg. min). Glycemia increased to 8.5 +/- 0.5 mmol/l at 60 min and remained elevated until the end of the glucose infusion (8.2 +/- 0.4 mmol/l). Together, our data show that the GLP-1 receptor is part of the hepatoportal glucose sensor and that basal fasting levels of GLP-1 sufficiently activate the receptor to confer maximum glucose competence to the sensor. These data demonstrate an important extrapancreatic effect of GLP-1 in the control of glucose homeostasis. PMID- 11473031 TI - T-cell epitope analysis on the autoantigen phogrin (IA-2beta) in the nonobese diabetic mouse. AB - The protein tyrosine phosphatases (PTPs) IA-2 and phogrin (IA-2beta) are major autoantigens in type 1 diabetes that possess common serological epitopes in their COOH termini. The epitopes recognized by the T-cells that cause the disease, however, remain to be defined. Eight phogrin-specific T-cell clones were generated from NOD mice, and their epitopes were mapped. The mapping was performed initially with recombinant gluthathione S-transferase-phogrin COOH deletion constructs and ultimately with overlapping synthetic peptides. Two dominant epitopes were identified: one (aa 629-649) immediately adjacent to the transmembrane domain (aa 604-628) and the second (aa 755-777) lying in the NH(2) terminal region of the conserved PTP domain. T-cells that are specific to either of these peptides and that could destroy islet tissue in vivo though spontaneous T-cell proliferative responses were observed in prediabetic female NOD splenocytes only to the aa 755-777 epitope. In NOD female mice immunized with the epitope peptide, intramolecular determinant spreading occurred from the aa 629 649 epitope to the aa 755-777 epitope but not in the opposite direction. We concluded that the initial T-cell response to phogrin is restricted to a small number of dominant peptides and that it subsequently spreads to other regions of the molecule, including those containing the major humoral epitopes that are highly conserved between IA-2 and phogrin. PMID- 11473032 TI - Expression of GAD65 and islet cell antibody (ICA512) autoantibodies among cytoplasmic ICA+ relatives is associated with eligibility for the Diabetes Prevention Trial-Type 1. AB - More than 71,000 relatives of type 1 diabetic patients have been screened for cytoplasmic islet cell antibodies (ICAs), GAD65 autoantibodies (GAAs), and ICA512 autoantibodies (ICA512AAs). Among those 71,148 relatives, 2,448 were cytoplasmic ICA+, and the remainder were ICA-. Of the ICA+ group, 1,229 (50.2%) were positive for GAAs and/or ICA512AAs. Among ICA- relatives, 1,897 (2.76%) were positive for GAAs and/or ICA512AAs. Given the large number of relatives positive for cytoplasmic ICA and negative for "biochemically" determined autoantibodies, and the converse, we analyzed the proportion of ICA+ relatives found eligible to participate in the intervention phase of Diabetes Prevention Trial-Type 1 (DPT 1). To be eligible for the parenteral insulin DPT-1 trial, a relative had to have first-phase insulin secretion below the 1st percentile of cut-points (for parents) or below the 10th percentile (for siblings and offspring). To be eligible for the oral insulin trial, a relative had to have first-phase insulin secretion above cut-points (>1st percentile for parents, >10th percentile for siblings/offspring) and be positive for anti-insulin autoantibodies. For both trials, DQB1*0602 was an exclusion criteria, cytoplasmic ICA positivity had to be confirmed, and an oral glucose tolerance test had to result in nondiabetic levels. Of 572 relatives found to be eligible for trial entry, 442 (77.3%) were positive for GAAs and/or ICA512AAs, although overall only 50.2% of ICA+ relatives were positive for GAAs and/or ICA512AAs. The positive predictive value for trial eligibility for ICA+ relatives with GAAs or ICA512AAs who completed staging was 51.0%. In contrast, only 11.9% of ICA+ but GAA- and ICA512AA- relatives were found to be eligible by DPT criteria for trial entry. Positivity for biochemically determined autoantibodies among cytoplasmic antibody-positive relatives is associated with eligibility for the DPT-1 study. PMID- 11473033 TI - Cytokine induction of Fas gene expression in insulin-producing cells requires the transcription factors NF-kappaB and C/EBP. AB - Fas-mediated cell death may play a role in the autoimmune destruction of pancreatic beta-cells in type 1 diabetes. beta-Cells do not express Fas under physiological conditions, but Fas mRNA and protein are induced in cytokine exposed mouse and human islets, rendering the beta-cells susceptible to Fas ligand-induced apoptosis. The aim of the present study was to investigate the molecular regulation of Fas by cytokines in rat beta-cells and in insulin producing RINm5F cells. Fas mRNA expression was increased 15-fold in fluorescence activated cell sorting-purified rat beta-cells exposed to interleukin (IL)-1beta, whereas gamma-interferon had no effect. Transfection experiments of rat Fas promoter-luciferase reporter constructs into purified rat beta-cells and RINm5F insulinoma cells identified an IL-1beta-responsive region between nucleotides 223 and -54. Inactivation of two adjacent NF-kappaB and C/EBP sites in this region abolished IL-1beta-induced Fas promoter activity in RINm5F cells. Binding of NF-kappaB and C/EBP factors to their respective sites was confirmed by gel shift assays. In cotransfection experiments, NF-kappaB p65 transactivated the Fas promoter. NF-kappaB p50 and C/EBPbeta overexpression had no effect by themselves on the Fas promoter activity, but when cotransfected with p65, each factor inhibited transactivation by p65. These results suggest a critical role for NF kappaB and C/EBP factors in cytokine-regulation of Fas expression in insulin producing cells. PMID- 11473034 TI - Characterization of preparations of GAD65, proinsulin, and the islet tyrosine phosphatase IA-2 for use in detection of autoreactive T-cells in type 1 diabetes: report of phase II of the Second International Immunology of Diabetes Society Workshop for Standardization of T-cell assays in type 1 diabetes. AB - The identification, quantification, and characterization of T-cells reactive with the islet autoantigens GAD65, proinsulin (PI), and tyrosine phosphatase-like molecules IA-2 and phogrin are major research goals in type 1 diabetes. In the Immunology of Diabetes Society First Workshop on Autoreactive T-Cells, the quality of recombinant preparations of these autoantigens was identified as a significant weakness, a finding that may account for much of the inconsistency in published studies of peripheral blood T-cell reactivity to islet autoantigens. Poor antigen quality has also hampered the development of novel technologies for the detection of islet-reactive T-cells. For these reasons, in the present study, several preparations of GAD65, PI, and IA-2 were collected and evaluated for endotoxin content, ability to stimulate a panel of relevant T-cell clones, and inhibitory effects on proliferation to unrelated third-party antigens. Through this process, we have been able to identify preparations of GAD65 and IA-2, generated in insect cells using the baculovirus expression system, that stimulate relevant clones and display low inhibitory effects on third-party antigens. In addition, we characterized a PI preparation generated in bacteria as being free of effects on proliferation to third-party antigens and low in endotoxin content. These preparations are important to promote the development of robust and sensitive assays of islet-reactive T-cells in patients with type 1 diabetes or patients at high risk for developing the disease. PMID- 11473035 TI - Nerve growth factor increases insulin secretion and barium current in pancreatic beta-cells. AB - We analyzed the effect of a brief exposure to nerve growth factor (NGF) on insulin secretion and macroscopic barium currents of single adult rat pancreatic beta-cells. After a 1-h exposure to NGF (50 ng/ml), single beta-cells show a 2.5 fold increase in the insulin secretion index in 5.6 mmol/l glucose and a nearly twofold increase in 15.6 mmol/l glucose compared with control cells. We have recently demonstrated that pancreatic beta-cells synthesize and secrete NGF. We analyzed the effect of endogenous NGF on insulin secretion by incubating islet cells in the presence of an anti-NGF monoclonal antibody for 1 h in different glucose concentrations. Although the basal insulin secretion index (5.6 mmol/l glucose) is not affected, glucose-stimulated insulin secretion (15.6 mmol/l glucose) is decreased by 41% in the presence of the antibody. This effect is mediated by the activation of the NGF receptor TrkA because the specific inhibitor of Trk phosphorylation K252a also blocks NGF-induced increase in insulin secretion, both in the presence and absence of exogenous NGF. Using the whole-cell variation of the patch-clamp technique, we found that cells exposed to NGF for 5 min exhibit a 32% increase in the average barium current density. These results suggest that the effects of NGF on insulin secretion are partially mediated by an increase in calcium current through Ca channels. These results further suggest that NGF plays an important autoregulatory role in pancreatic beta-cell function. Two targets of short-term NGF-modulation are insulin secretion and calcium-channel activity. PMID- 11473036 TI - Linear correlation between the total islet mass and the volume-weighted mean islet volume. AB - To understand the dynamics of islet population, especially during conditions with growth of the total islet mass, it is important to have reliable estimators of parameters describing the quantitative appearance of the islet population. We describe a stereological estimator of the volume-weighted mean islet volume based on unbiased assumption-free stereological principles. The volume-weighted mean islet volume is the mean islet volume if the islets are weighted (sampled) proportional to their volume. This method allows simultaneously unbiased estimation of the total islet mass. With use of this method, 22 male Sprague Dawley rats within the age span of 34-102 days old were investigated. We found a linear correlation (P < 0.001) between total islet mass and the volume-weighted mean islet volume. The results support models demonstrating that the physiological growth of the total islet mass in the period studied is totally or mainly caused by proportional growth of existing islets. The functional meaning of the volume-weighted mean islet volume is discussed, and previous methods to study the mean islet volume and islet number are critically evaluated. We propose the volume-weighted mean islet volume to be a biologically useful parameter when describing the mean volume of the pancreatic islets and investigating the differences between experimental groups. PMID- 11473037 TI - Inverse relationship between cytotoxicity of free fatty acids in pancreatic islet cells and cellular triglyceride accumulation. AB - Studies in Zucker diabetic fatty rats have led to the concept that chronically elevated free fatty acid (FFA) levels can cause apoptosis of triglyceride-laden pancreatic beta-cells as a result of the formation of ceramides, which induce nitric oxide (NO)-dependent cell death. This "lipotoxicity" hypothesis could explain development of type 2 diabetes in obesity. The present study examines whether prolonged exposure to FFA affects survival of isolated normal rat beta cells and whether the outcome is related to the occurrence of triglyceride accumulation. A dose-dependent cytotoxicity was detected at 5-100 nmol/l of unbound oleate and palmitate, with necrosis occurring within 48 h and an additional apoptosis during the subsequent 6 days of culture. At equimolar concentrations, the cytotoxicity of palmitate was higher than that of oleate but lower than that of its nonmetabolized analog bromopalmitate. FFA cytotoxicity was not suppressed by etomoxir (an inhibitor of mitochondrial carnitine palmitoyltransferase I) or by antioxidants; it was not associated with inducible NO synthase expression or NO formation. An inverse correlation was observed between the percentage of dead beta-cells on day 8 and their cellular triglyceride content on day 2. For equimolar concentrations of the tested FFA, oleate caused the lowest beta-cell toxicity and the highest cytoplasmic triglyceride accumulation. On the other hand, oleate exerted the highest toxicity in islet non-beta-cells, where no FFA-induced triglyceride accumulation was detected. In conditions without triglyceride accumulation, the lower FFA concentrations caused primarily apoptosis, both in islet beta-cells and non-beta cells. It is concluded that FFAs can cause death of normal rat islet cells through an NO-independent mechanism. The ability of normal beta-cells to form and accumulate cytoplasmic triglycerides might serve as a cytoprotective mechanism against FFA-induced apoptosis by preventing a cellular rise in toxic free fatty acyl moieties. It is conceivable that this potential is lost or insufficient in cells with a prolonged triglyceride accumulation as may occur in vivo. PMID- 11473038 TI - Acute and short-term administration of a sulfonylurea (gliclazide) increases pulsatile insulin secretion in type 2 diabetes. AB - The high-frequency oscillatory pattern of insulin release is disturbed in type 2 diabetes. Although sulfonylurea drugs are widely used for the treatment of this disease, their effect on insulin release patterns is not well established. The aim of the present study was to assess the impact of acute treatment and 5 weeks of sulfonylurea (gliclazide) treatment on insulin secretory dynamics in type 2 diabetic patients. To this end, 10 patients with type 2 diabetes (age 53 +/- 2 years, BMI 27.5 +/- 1.1 kg/m(2), fasting plasma glucose 9.8 +/- 0.8 mmol/l, HbA(1c) 7.5 +/- 0.3%) were studied in a double-blind placebo-controlled prospective crossover design. Patients received 40-80 mg gliclazide/placebo twice daily for 5 weeks with a 6-week washout period intervening. Insulin pulsatility was assessed by 1-min interval blood sampling for 75 min 1) under baseline conditions (baseline), 2) 3 h after the first dose (80 mg) of gliclazide (acute) with the plasma glucose concentration clamped at the baseline value, 3) after 5 weeks of treatment (5 weeks), and 4) after 5 weeks of treatment with the plasma glucose concentration clamped during the sampling at the value of the baseline assessment (5 weeks-elevated). Serum insulin concentration time series were analyzed by deconvolution, approximate entropy (ApEn), and spectral and autocorrelation methods to quantitate pulsatility and regularity. The P values given are gliclazide versus placebo; results are means +/- SE. Fasting plasma glucose was reduced after gliclazide treatment (baseline vs. 5 weeks: gliclazide, 10.0 +/- 0.9 vs. 7.8 +/- 0.6 mmol/l; placebo, 10.0 +/- 0.8 vs. 11.0 +/- 0.9 mmol/l, P = 0.001). Insulin secretory burst mass was increased (baseline vs. acute: gliclazide, 43.0 +/- 12.0 vs. 61.0 +/- 17.0 pmol. l(-1). pulse(-1); placebo, 36.1 +/- 8.4 vs. 30.3 +/- 7.4 pmol. l(-1). pulse(-1), P = 0.047; 5 weeks elevated: gliclazide vs. placebo, 49.7 +/- 13.3 vs. 37.1 +/- 9.5 pmol. l(-1). pulse(-1), P < 0.05) with a similar rise in burst amplitude. Basal (i.e., nonoscillatory) insulin secretion also increased (baseline vs. acute: gliclazide, 8.5 +/- 2.2 vs. 16.7 +/- 4.3 pmol. l(-1). pulse(-1); placebo, 5.9 +/- 0.9 vs. 7.2 +/- 0.9 pmol. l(-1). pulse(-1), P = 0.03; 5 weeks-elevated: gliclazide vs. placebo, 12.2 +/- 2.5 vs. 9.4 +/- 2.1 pmol. l(-1). pulse(-1), P = 0.016). The frequency and regularity of insulin pulses were not modified significantly by the antidiabetic therapy. There was, however, a correlation between individual values for the acute improvement of regularity, as measured by ApEn, and the decrease in fasting plasma glucose during short-term (5-week) gliclazide treatment (r = 0.74, P = 0.014, and r = 0.77, P = 0.009, for fine and coarse ApEn, respectively). In conclusion, the sulfonylurea agent gliclazide augments insulin secretion by concurrently increasing pulse mass and basal insulin secretion without changing secretory burst frequency or regularity. The data suggest a possible relationship between the improvement in short-term glycemic control and the acute improvement of regularity of the in vivo insulin release process. PMID- 11473039 TI - Glucose metabolism and pulsatile insulin release from isolated islets. AB - The effects of metabolic inhibition on insulin release and the cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) were studied in individually perifused pancreatic islets from ob/ob mice. The modest basal secretion in the presence of 3 mmol/l glucose was pulsatile with a frequency of approximately 0.2/min, although [Ca(2+)](i) was stable at approximately 100 nmol/l. Introduction of 11 mmol/l glucose resulted in large amplitude oscillations of [Ca(2+)](i) and almost 20 fold stimulation of average secretion manifested as increased amplitude of the insulin pulses without change in frequency. Inhibition of glycolysis with iodoacetamide or mitochondrial metabolism with dinitrophenol or antimycin A reduced glucose-stimulated secretion back to basal levels with maintained pulsatility. The [Ca(2+)](i) responses to the metabolic inhibitors were more complex, but in general there was an initial peak and eventually sustained elevation without oscillations. When introduced in the presence of 3 mmol/l glucose, the metabolic inhibitors tended to increase the amplitude of the insulin pulses, although the simultaneous elevation in [Ca(2+)](i) occurred without oscillations. The data indicate that pulsatile secretion is regulated by factors other than [Ca(2+)](i) under basal conditions and after metabolic inhibition. Although pulsatile secretion can be driven by oscillations in metabolism when [Ca(2+)](i) is stable, it was not possible from the present data to determine whether insulin pulses have a glycolytic or mitochondrial origin. PMID- 11473040 TI - Glycerol-stimulated proinsulin biosynthesis in isolated pancreatic rat islets via adenoviral-induced expression of glycerol kinase is mediated via mitochondrial metabolism. AB - In this study, we examined whether adenoviral-mediated glycerol kinase (AdV-CMV GlyK) expression in isolated rat pancreatic islets could introduce glycerol induced proinsulin biosynthesis. In AdV-CMV-GlyK-infected islets, specific glycerol-induced proinsulin biosynthesis translation and insulin secretion were observed in parallel from the same islets. The threshold concentration of glycerol required to stimulate proinsulin biosynthesis was lower (0.25-0.5 mmol/l) than that for insulin secretion (1.0-1.5 mmol/l), reminiscent of threshold differences for glucose-stimulated proinsulin biosynthesis versus insulin secretion. The dose-dependent glycerol-induced proinsulin biosynthesis correlated with the rate of glycerol oxidation in AdV-CMV-GlyK-infected islets, indicating that glycerol metabolism was required for the response. However, glycerol did not significantly increase lactate output from AdV-CMV-GlyK-infected islets, but the dihydroxyacetone phosphate (DHAP) to alpha-glycerophosphate (alpha-GP) ratio significantly increased in AdV-CMV-GlyK-infected islets incubated at 2 mmol/l glycerol compared with that at a basal level of 2.8 mmol/l glucose (P < or = 0.05). The DHAP:alpha-GP ratio was unaffected in AdV-CMV-GlyK infected islets incubated at 2 mmol/l glycerol in the added presence of alpha cyanohydroxycinnaminic acid (alpha-CHC), an inhibitor of the plasma membrane and mitochondrial lactate/pyruvate transporter. However, alpha-CHC inhibited glycerol induced proinsulin biosynthesis and insulin secretion in AdV-CMV-GlyK-infected islets (>75%; P = 0.05), similarly to glucose-induced proinsulin biosynthesis and insulin secretion in AdV-CMV-GlyK-infected and control islets. These data indicated that in AdV-CMV-GlyK-infected islets, the importance of mitochondrial metabolism of glycerol was required to generate stimulus-response coupling signals to induce proinsulin biosynthesis and insulin secretion. PMID- 11473041 TI - IPF1/PDX1 deficiency and beta-cell dysfunction in Psammomys obesus, an animal With type 2 diabetes. AB - The homeodomain transcription factor IPF1/PDX1 is required in beta-cells for efficient expression of insulin, glucose transporter 2, and prohormone convertases 1/3 and 2. Psammomys obesus, a model of diet-responsive type 2 diabetes, shows markedly depleted insulin stores when given a high-energy (HE) diet. Despite hyperglycemia, insulin mRNA levels initially remained unchanged and then decreased gradually to 15% of the basal level by 3 weeks. Moreover, insulin gene expression was not increased when isolated P. obesus islets were exposed to elevated glucose concentrations. Consistent with these observations, no functional Ipf1/Pdx1 gene product was detected in islets of newborn or adult P. obesus using immunostaining, Western blot, DNA binding, and reverse transcriptase polymerase chain reaction analyses. Other beta-cell transcription factors (e.g., ISL-1, Nkx2.2, and Nkx6.1) were expressed in P. obesus islets, and the DNA binding activity of the insulin transcription factors RIPE3b1-Act and IEF1 was intact. Ipf1/Pdx1 gene transfer to isolated P. obesus islets normalized the defect in glucose-stimulated insulin gene expression and prevented the rapid depletion of insulin content after exposure to high glucose. Taken together, these results suggest that the inability of P. obesus islets to adapt to dietary overload, with depletion of insulin content as a consequence, results from IPF1/PDX1 deficiency. However, because not all animals become hyperglycemic on HE diet, additional factors may be important for the development of diabetes in this animal model. PMID- 11473042 TI - Effect of physiological hyperinsulinemia on gluconeogenesis in nondiabetic subjects and in type 2 diabetic patients. AB - Gluconeogenesis (GNG) is enhanced in type 2 diabetes. In experimental animals, insulin at high doses decreases the incorporation of labeled GNG precursors into plasma glucose. Whether physiological hyperinsulinemia has any effect on total GNG in humans has not been determined. We combined the insulin clamp with the (2)H(2)O technique to measure total GNG in 33 subjects with type 2 diabetes (BMI 29.0 +/- 0.6 kg/m(2), fasting plasma glucose 8.1 +/- 0.3 mmol/l) and in 9 nondiabetic BMI-matched subjects after 16 h of fasting and after euglycemic hyperinsulinemia. A primed-constant infusion of 6,6-(2)H-glucose was used to monitor endogenous glucose output (EGO); insulin (40 mU. min(-1). m(-2)) was then infused while clamping plasma glucose for 2 h (at 5.8 +/- 0.1 and 4.9 +/- 0.2 mmol/l for diabetic and control subjects, respectively). In the fasting state, EGO averaged 15.2 +/- 0.4 micromol. min(-1). kg(-1)(ffm) (62% from GNG) in diabetic subjects and 12.2 +/- 0.7 micromol. min(-1). kg(-1)(ffm) (55% from GNG) in control subjects (P < 0.05 or less for both fluxes). Glycogenolysis (EGO - GNG) was similar in the two groups (P = NS). During the last 40 min of the clamp, both EGO and GNG were significantly (P < 0.01 or less, compared with fasting) inhibited (EGO 7.1 +/- 0.9 and 3.6 +/- 0.5 and GNG 7.9 +/- 0.5 and 4.5 +/- 1.0 respectively) but remained significantly (P < 0.05) higher in diabetic subjects, whereas glycogenolysis was suppressed completely and equally in both groups. During hyperinsulinemia, GNG micromol. min(-1). kg(-1)(ffm) in diabetic and control subjects, was reciprocally related to plasma glucose clearance. In conclusion, physiological hyperinsulinemia suppresses GNG by approximately 20%, while completely blocking glycogenolysis. Resistance of GNG (to insulin suppression) and resistance of glucose uptake (to insulin stimulation) are coupled phenomena. In type 2 diabetes, the excess GNG of the fasting state is carried over to the insulinized state, thereby contributing to glucose overproduction under both conditions. PMID- 11473043 TI - Treatment of type 2 diabetes by adenoviral-mediated overexpression of the glucokinase regulatory protein. AB - The enzyme glucokinase (GK) plays a central role in glucose homeostasis. Hepatic GK activity is acutely controlled by the action of the GK regulatory protein (GKRP). In vitro evidence suggests that GKRP reversibly binds to GK and inhibits its activity; however, less is known about the in vivo function of GKRP. To further explore the physiological role of GKRP in vivo, we used an E1/E2a/E3 deficient adenoviral vector containing the cDNA encoding human GKRP (Av3hGKRP). High fat diet-induced diabetic mice were administered Av3hGKRP or a control vector lacking a transgene (Av3Null). Surprisingly, the Av3hGKRP-treated mice showed a significant improvement in glucose tolerance and had lower fasting blood glucose levels than Av3Null-treated mice. A coincident decrease in insulin levels indicated that the Av3hGKRP-treated mice had sharply improved insulin sensitivity. These mice also exhibited lower leptin levels, reduced body weight, and decreased liver GK activity. In vitro experiments indicated that GKRP was able to increase both GK protein and enzymatic activity levels, suggesting that another role for GKRP is to stabilize and/or protect GK. These data are the first to indicate the ability of GKRP to treat type 2 diabetes and therefore have significant implications for future therapies of this disease. PMID- 11473044 TI - Alteration in phosphorylation of P20 is associated with insulin resistance. AB - We have recently identified a small phosphoprotein, P20, as a common intracellular target for insulin and several of its antagonists, including amylin, epinephrine, and calcitonin gene-related peptide. These hormones elicit phosphorylation of P20 at its different sites, producing three phosphorylated isoforms: S1 with an isoelectric point (pI) value of 6.0, S2 with a pI value of 5.9, and S3 with a pI value of 5.6 (FEBS Letters 457:149-152 and 462:25-30, 1999). In the current study, we showed that P20 is one of the most abundant phosphoproteins in rat extensor digitorum longus (EDL) muscle. Insulin and amylin antagonize each other's actions in the phosphorylation of this protein in rat EDL muscle. Insulin inhibits amylin-evoked phosphorylation of S2 and S3, whereas amylin decreases insulin-induced phosphorylation of S1. In rats made insulin resistant by dexamethasone treatment, levels of the phosphoisoforms S2 and S3, which were barely detectable in healthy rats in the absence of hormone stimulation, were significantly increased. Moreover, the ability of insulin to inhibit amylin-evoked phosphorylation of these two isoforms was greatly attenuated. These results suggested that alterations in the phosphorylation of P20 might be associated with insulin resistance and that P20 could serve as a useful marker to dissect the cellular mechanisms of this disease. PMID- 11473045 TI - Low acute insulin secretory responses in adult offspring of people with early onset type 2 diabetes. AB - The offspring of Pima Indians with early onset type 2 diabetes are at high risk for developing diabetes at an early age. This risk is greater among those whose mothers were diabetic during pregnancy. To define the metabolic abnormalities predisposing individuals in these high-risk groups to diabetes, we conducted a series of studies to measure insulin secretion and insulin action in healthy adult Pima Indians. In 104 normal glucose-tolerant subjects, acute insulin secretory response (AIR) to a 25-g intravenous glucose challenge correlated with the age at onset of diabetes in the mother (r = 0.23, P = 0.03) and, in multiple regression analyses, the age at onset of diabetes in the father (P = 0.02), after adjusting for maternal age at onset and after allowing for an interaction between these terms. In contrast, insulin action (hyperinsulinemic glucose clamp) did not correlate with the age at onset of diabetes in the parents. To determine whether early onset diabetes in the parents affected insulin secretion in the offspring across a range of glucose concentrations, responses to a stepped glucose infusion were measured in 23 subjects. Insulin secretion rates were lower in individuals whose mothers had developed diabetes before 35 years of age (n = 8) compared with those whose parents remained nondiabetic until at least 49 years of age (n = 15) (average insulin secretory rates: geometric mean [95% CI] 369 [209-652] vs. 571 [418-780] pmol/min, P = 0.007). Finally, the AIR was lower in individuals whose mothers were diabetic during pregnancy (n = 8) than in those whose mothers developed diabetes at an early age but after the birth of the subject (n = 41) (740 [510-1,310] vs. 1,255 [1,045-1,505] pmol/l, P < 0.02). Thus, insulin secretion is lower in normal glucose tolerant offspring of people with early onset type 2 diabetes. This impairment may be worsened by exposure to a diabetic environment in utero. PMID- 11473046 TI - Insulin secretion and glucose kinetics during exercise with and without pharmacological alpha(1)- and alpha(2)-receptor blockade. AB - The mechanism behind exercise-induced decreases in plasma insulin concentrations was examined in eight healthy young men. In addition, the influence of specific alpha(1)- and alpha(2)-adrenoceptor blockade on glucose kinetics during exercise was studied. To test the hypothesis that exercise-induced decreases in insulin secretion are mediated via alpha(2)-adrenoceptors, all subjects exercised for 60 min on separate occasions under four conditions: with and without alpha(1) receptor blockade (1 mg prazosin) and with and without or alpha(2)-receptor blockade (15 mg yohimbine). Glucose kinetics were measured using [3-(3)H]glucose. During exercise with alpha(2)-receptor blockade, the insulin concentration initially increased (first 20 min) then decreased, whereas it continually decreased in the corresponding control experiment. The C-peptide concentration did not change during exercise with alpha(2)-receptor blockade but decreased in the control experiment. During exercise with alpha(1)-receptor blockade and corresponding control experiments, insulin and C-peptide levels always decreased. With alpha(1)-receptor blockade, the glucose concentration increased (first 30 min) and then decreased, whereas it slightly decreased in all other experiments. In addition, with alpha(1)-receptor blockade, the glucose rate of appearance (Ra) increased rapidly (because of higher catecholamine concentrations in alpha(1) receptor blockade versus control) and the glucose rate of disappearance (Rd) was higher compared with control. During exercise with alpha(2)-receptor blockade, the Ra and Rd were always lower compared with control. Therefore, we conclude that exercise-induced decreases in insulin secretion are mediated via alpha(2) adrenoceptors and that blockade of alpha(1)- and alpha(2)-adrenoceptors during exercise elicits opposite responses in glucose Ra and Rd. PMID- 11473047 TI - Nonalcoholic fatty liver disease: a feature of the metabolic syndrome. AB - Insulin sensitivity (euglycemic clamp, insulin infusion rate: 40 mU. m(-2). min( 1)) was studied in 30 subjects with biopsy-proven nonalcoholic fatty liver disease (NAFLD), normal glucose tolerance, and a BMI <30 kg/m(2). Of those 30 subjects, 9 had pure fatty liver and 21 had evidence of steatohepatitis. In addition, 10 patients with type 2 diabetes under good metabolic control and 10 healthy subjects were studied. Most NAFLD patients had central fat accumulation, increased triglycerides and uric acid, and low HDL cholesterol, irrespective of BMI. Glucose disposal during the clamp was reduced by nearly 50% in NAFLD patients, as well as in patients with normal body weight, to an extent similar to that of the type 2 diabetic patients. Basal free fatty acids were increased, whereas insulin-mediated suppression of lipolysis was less effective (-69% in NAFLD vs. -84% in control subjects; P = 0.003). Postabsorptive hepatic glucose production (HGP), measured by [6,6-(2)H(2)]glucose, was normal. In response to insulin infusion, HGP decreased by only 63% of basal in NAFLD vs. 84% in control subjects (P = 0.002). Compared with type 2 diabetic patients, NAFLD patients were characterized by lower basal HGP, but with similarly reduced insulin-mediated suppression of HGP. There was laboratory evidence of iron overload in many NAFLD patients, but clinical, histological, and biochemical data (including insulin sensitivity) were not correlated with iron status. Four subjects were heterozygous for mutation His63Asp of the HFE gene of familiar hemochromatosis. We concluded that NAFLD, in the presence of normoglycemia and normal or moderately increased body weight, is characterized by clinical and laboratory data similar to those found in diabetes and obesity. NAFLD may be considered an additional feature of the metabolic syndrome, with specific hepatic insulin resistance. PMID- 11473048 TI - Glucose regulates the transcription of human genes relevant to HDL metabolism: responsive elements for peroxisome proliferator-activated receptor are involved in the regulation of phospholipid transfer protein. AB - Phospholipid transfer protein (PLTP) plays an important role in human plasma HDL metabolism. Clinical data have recently indicated that plasma PLTP activity and mass were both higher in diabetic patients concomitant with hyperglycemia. The present study shows that high glucose increases both PLTP mRNA and functional activity in HepG2 cells, due to a significant increase in the promoter activity of human PLTP gene. The glucose-responsive elements are located between -759 and 230 of the PLTP 5'-flanking region, within which two binding motifs (-537 to -524 and -339 to -327) for either peroxisome proliferator-activated receptor or farnesoid X-activated receptor are involved in this glucose-mediated transcriptional regulation. This finding suggests that high glucose upregulates the transcription of human PLTP gene via nuclear hormone receptors. In addition, high glucose increases mRNA levels for several genes that are functionally important in HDL metabolism, including human ATP-binding cassette transporter A1, apolipoprotein A-I, scavenger receptor BI, and hepatic lipase. The functional promoter activities of these genes are enhanced by high glucose in three cell lines tested, indicating that glucose may also regulate these genes at the transcriptional level. Our findings provide a molecular basis for a role of hyperglycemia in altered HDL metabolism. PMID- 11473049 TI - Menopause in type 1 diabetic women: is it premature? AB - Women with type 1 diabetes have a delayed menarche and a greater prevalence of menstrual disorders than women without diabetes. However, little is known about the menopause transition among type 1 diabetic women. The Familial Autoimmune and Diabetes (FAD) Study recruited both adult individuals who were identified from the Children's Hospital of Pittsburgh Type 1 Diabetes Registry for the years 1950 1964 and their family members. Unrelated nondiabetic control probands and their relatives were also evaluated. Women with type 1 diabetes (n = 143) compared with nondiabetic sisters (n = 186) or unrelated control subjects (n = 160) were more likely to have an older age at menarche (13.5, 12.5, and 12.6 years, respectively, P < 0.001), more menstrual irregularities before 30 years of age (45.7, 33.3, and 33.1%, respectively, P = 0.04), and a younger age at menopause (41.6, 49.9, and 48.0 years, respectively, P = 0.05). This resulted in a 6-year reduction in the number of reproductive years (30.0, 37.0, and 35.2 years, respectively, P = 0.05) for women with type 1 diabetes. Risk factors univariately associated with earlier menopause included type 1 diabetes (hazard ratio [HR] 1.99, P = 0.04), menstrual irregularities before 30 years of age (HR 1.87, P = 0.04), nulliparity (HR 2.14, P = 0.01), and unilateral oophorectomy (HR 6.51, P < 0.0001). Multivariate analysis confirmed that type 1 diabetes (HR 1.98, P = 0.056), menstrual irregularities by 30 years of age (HR 2.36, P = 0.01), and unilateral oophorectomy (HR 9.76, P < 0.0001) were independent determinants of earlier menopause in our cohort. We hypothesize that an earlier menopause, which resulted in a 17% decrease in reproductive years, is a major unstudied complication of type 1 diabetes. PMID- 11473050 TI - Effects of pioglitazone on adipose tissue remodeling within the setting of obesity and insulin resistance. AB - Obesity and dysfunctional energy partitioning can lead to the development of insulin resistance and type 2 diabetes. The antidiabetic thiazolidinediones shift the energy balance toward storage, leading to an increase in whole-body adiposity. These studies examine the effects of pioglitazone (Pio) on adipose tissue physiology, accumulation, and distribution in female Zucker (fa/fa) rats. Pio treatment (up to 28 days) decreased the insulin-resistant and hyperlipidemic states and increased food consumption and whole-body adiposity. Magnetic resonance imaging (MRI) analysis and weights of fat pads demonstrated that the increase in adiposity was not only limited to the major fat depots but also to fat deposition throughout the body. Adipocyte sizing profiles, fat pad histology, and DNA content show that Pio treatment increased the number of small adipocytes because of both the appearance of new adipocytes and the shrinkage and/or disappearance of existing mature adipocytes. The remodeling was time dependent, with new small adipocytes appearing in clusters throughout the fat pad, and accompanied by a three- to fourfold increase in citrate synthase and fatty acid synthase activity. The appearance of new fat cells and the increase in fat mass were depot specific, with a rank order of responsiveness of ovarian > retroperitoneal > subcutaneous. This differential depot effect resulted in a redistribution of the fat mass in the abdominal region such that there was an increase in the visceral:subcutaneous ratio, as confirmed by MRI analysis. Although the increased adiposity is paradoxical to an improvement in insulin sensitivity, the quantitative increase of adipose mass should be viewed in context of the qualitative changes in adipose tissue, including the remodeling of adipocytes to a smaller size with higher lipid storage potential. This shift in energy balance is likely to result in lower circulating free fatty acid levels, ultimately improving insulin sensitivity and the metabolic state. PMID- 11473051 TI - Small increases in insulin inhibit hepatic glucose production solely caused by an effect on glycogen metabolism. AB - Based on our earlier work, a 2.5-fold increase in insulin secretion should completely inhibit hepatic glucose production through the hormone's direct effect on hepatic glycogen metabolism. The aim of the present study was to test the accuracy of this prediction and to confirm that gluconeogenic flux, as measured by three independent techniques, was unaffected by the increase in insulin. A 40 min basal period was followed by a 180-min experimental period in which an increase in insulin was induced, with euglycemia maintained by peripheral glucose infusion. Arterial and hepatic sinusoidal insulin levels increased from 10 +/- 2 to 19 +/- 3 and 20 +/- 4 to 45 +/- 5 microU/ml, respectively. Net hepatic glucose output decreased rapidly from 1.90 +/- 0.13 to 0.23 +/- 0.16 mg. kg(-1). min(-1). Three methods of measuring gluconeogenesis and glycogenolysis were used: 1) the hepatic arteriovenous difference technique (n = 8), 2) the [(14)C]phosphoenolpyruvate technique (n = 4), and 3) the (2)H(2)O technique (n = 4). The net hepatic glycogenolytic rate decreased from 1.72 +/- 0.20 to -0.28 +/- 0.15 mg. kg(-1). min(-1) (P < 0.05), whereas none of the above methods showed a significant change in hepatic gluconeogenic flux (rate of conversion of phosphoenolpyruvate to glucose-6-phosphate). These results indicate that liver glycogenolysis is acutely sensitive to small changes in plasma insulin, whereas gluconeogenic flux is not. PMID- 11473052 TI - Bezafibrate reduces mRNA levels of adipocyte markers and increases fatty acid oxidation in primary culture of adipocytes. AB - The molecular mechanisms by which peroxisome proliferator-activated receptor (PPAR) activation by fibrates reduces fat deposition and improves insulin sensitivity are not completely understood. We report that exposure of a rat primary culture of adipocytes for 24 h to the PPAR activator bezafibrate increased the mRNA levels of crucial genes involved in peroxisomal and mitochondrial beta-oxidation. The mRNA levels of the peroxisomal beta-oxidation rate-limiting enzyme acyl-CoA oxidase and of the muscle-type carnitine palmitoyl transferase I (M-CPT-I), which determines the flux of mitochondrial beta oxidation, increased by 1.6-fold (P < 0.02) and 4.5-fold (P = 0.001), respectively. These changes were accompanied by an increase in the transcript levels of the uncoupling protein-2 (UCP-2; 1.5-fold induction; P < 0.05) and UCP 3 (3.7-fold induction; P < 0.001), mitochondrial proteins that reduce ATP yield and may facilitate the oxidation of fatty acids. Furthermore, bezafibrate increased the mRNA levels of the fatty acid translocase (2-fold induction; P < 0.01), suggesting a higher fatty acid uptake into adipocytes. In agreement with these changes, bezafibrate caused a 1.9-fold induction (P < 0.02) in 9,10 [(3)H]palmitate oxidation. Moreover, bezafibrate reduced the mRNA expression of several adipocyte markers, including PPARgamma (30% reduction; P = 0.05), tumor necrosis factor-alpha (33% reduction; P < 0.05), and the ob gene (26% reduction). In contrast, adipocyte fatty acid binding protein mRNA levels increased (1.5-fold induction; P < 0.01), pointing to a mobilization of fatty acids to mitochondria and peroxisomes. The reduction of the adipocyte markers caused by bezafibrate was accompanied by an increase in the mRNA levels of the preadipocyte marker Pref-1 (1.6-fold induction; P < 0.01). Some of the changes observed in the primary culture of rat adipocytes also were studied in the epididymal white adipose tissue of bezafibrate-treated rats for 7 days. In vivo, M-CPT-I mRNA levels increased (4.5-fold induction; P = 0.001) in epididymal white adipose tissue of bezafibrate-treated rats. Similarly, fatty acid translocase (2.6-fold induction; P = 0.002) and Pref-1 (5.6-fold induction) mRNA levels increased, although differences in the latter were not significant because of huge individual variations. These results indicate that exposure of adipocytes to bezafibrate, independent of its hepatic effects, increases the degradation of fatty acids, reducing their availability to synthesize triglycerides. As a result, some degree of dedifferentiation of adipocytes to preadipocyte-like cells is achieved. These changes may be involved in the reduction in fat depots and in the improvement of insulin sensitivity observed after bezafibrate treatment. PMID- 11473053 TI - Growth hormone induces cellular insulin resistance by uncoupling phosphatidylinositol 3-kinase and its downstream signals in 3T3-L1 adipocytes. AB - Growth hormone (GH) is well known to induce in vivo insulin resistance. However, the molecular mechanism of GH-induced cellular insulin resistance is largely unknown. In this study, we demonstrated that chronic GH treatment of differentiated 3T3-L1 adipocytes reduces insulin-stimulated 2-deoxyglucose (DOG) uptake and activation of Akt (also known as protein kinase B), both of which are downstream effects of phosphatidylinositol (PI) 3-kinase, despite enhanced tyrosine phosphorylation of insulin receptor substrate (IRS)-1, association of IRS-1 with the p85 subunit of PI 3-kinase, and IRS-1-associated PI 3-kinase activity. In contrast, chronic GH treatment did not affect 2-DOG uptake and Akt activation induced by overexpression of a membrane-targeted form of the p110 subunit of PI 3-kinase (p110(CAAX)) or Akt activation stimulated by platelet derived growth factor. Fractionation studies indicated that chronic GH treatment reduces insulin-stimulated translocation of Akt from the cytosol to the plasma membrane. Interestingly, chronic GH treatment increased insulin-stimulated association of IRS-1 with p85 and IRS-1-associated PI 3-kinase activity preferentially in the cytosol. These results indicate that cellular insulin resistance induced by chronic GH treatment in 3T3-L1 adipocytes is caused by uncoupling between activation of PI 3-kinase and its downstream signals, which is specific to the insulin-stimulated PI 3-kinase pathway. This effect of GH might result from the altered subcellular distribution of IRS-1-associated PI 3-kinase. PMID- 11473054 TI - Defective insulin-induced GLUT4 translocation in skeletal muscle of high fat-fed rats is associated with alterations in both Akt/protein kinase B and atypical protein kinase C (zeta/lambda) activities. AB - The cellular mechanism by which high-fat feeding induces skeletal muscle insulin resistance was investigated in the present study. Insulin-stimulated glucose transport was impaired ( approximately 40-60%) in muscles of high fat-fed rats. Muscle GLUT4 expression was significantly lower in these animals ( approximately 40%, P < 0.05) but only in type IIa-enriched muscle. Insulin stimulated the translocation of GLUT4 to both the plasma membrane and the transverse (T)-tubules in chow-fed rats. In marked contrast, GLUT4 translocation was completely abrogated in the muscle of insulin-stimulated high fat-fed rats. High-fat feeding markedly decreased insulin receptor substrate (IRS)-1-associated phosphatidylinositol (PI) 3-kinase activity but not insulin-induced tyrosine phosphorylation of the insulin receptor and IRS proteins in muscle. Impairment of PI 3-kinase function was associated with defective Akt/protein kinase B kinase activity (-40%, P < 0.01) in insulin-stimulated muscle of high fat-fed rats, despite unaltered phosphorylation (Ser473/Thr308) of the enzyme. Interestingly, basal activity of atypical protein kinase C (aPKC) was elevated in muscle of high fat-fed rats compared with chow-fed controls. Whereas insulin induced a twofold increase in aPKC kinase activity in the muscle of chow-fed rats, the hormone failed to further increase the kinase activity in high fat-fed rat muscle. In conclusion, it was found that GLUT4 translocation to both the plasma membrane and the T-tubules is impaired in the muscle of high fat-fed rats. We identified PI 3 kinase as the first step of the insulin signaling pathway to be impaired by high fat feeding, and this was associated with alterations in both Akt and aPKC kinase activities. PMID- 11473055 TI - Blood-to-brain glucose transport, cerebral glucose metabolism, and cerebral blood flow are not increased after hypoglycemia. AB - Recent antecedent hypoglycemia has been found to shift glycemic thresholds for autonomic (including adrenomedullary epinephrine), symptomatic, and other responses to subsequent hypoglycemia to lower plasma glucose concentrations. This change in threshold is the basis of the clinical syndromes of hypoglycemia unawareness and, in part, defective glucose counterregulation and the unifying concept of hypoglycemia-associated autonomic failure in type 1 diabetes. We tested in healthy young adults the hypothesis that recent antecedent hypoglycemia increases blood-to-brain glucose transport, a plausible mechanism of this phenomenon. Eight subjects were studied after euglycemia, and nine were studied after approximately 24 h of interprandial hypoglycemia ( approximately 55 mg/dl, approximately 3.0 mmol/l). The latter were shown to have reduced plasma epinephrine (P = 0.009), neurogenic symptoms (P = 0.009), and other responses to subsequent hypoglycemia. Global bihemispheric blood-to-brain glucose transport and cerebral glucose metabolism were calculated from rate constants derived from blood and brain time-activity curves-the latter determined by positron emission tomography (PET)-after intravenous injection of [1-(11)C]glucose at clamped plasma glucose concentrations of 65 mg/dl (3.6 mmol/l). For these calculations, a model was used that includes a fourth rate constant to account for egress of [(11)C] metabolites. Cerebral blood flow was measured with intravenous [(15)O]water using PET. After euglycemia and after hypoglycemia, rates of blood to-brain glucose transport (24.6 +/- 2.3 and 22.4 +/- 2.4 micromol. 100 g(-1). min(-1), respectively), cerebral glucose metabolism (16.8 +/- 0.9 and 15.9 +/- 0.9 micromol. 100 g(-1). min(-1), respectively) and cerebral blood flow (56.8 +/- 3.9 and 53.3 +/- 4.4 ml. 100 g(-1). min(-1), respectively) were virtually identical. These data do not support the hypothesis that recent antecedent hypoglycemia increases blood-to-brain glucose transport during subsequent hypoglycemia. They do not exclude regional increments in blood-to-brain glucose transport. Alternatively, the fundamental alteration might lie beyond the blood brain barrier. PMID- 11473056 TI - Angiotensin II promotes glucose-induced activation of cardiac protein kinase C isozymes and phosphorylation of troponin I. AB - Activation of the protein kinase C (PKC) family is a potential signaling mechanism by which high ambient glucose concentration modulates the phenotype and physiological function of cells. Recently, the cardiac renin angiotensin system (RAS) has been reported to promote PKC translocation in the diabetic heart via the angiotensin (ANG) II type 1 receptor (AT-1R). To evaluate the molecular events coupled with high glucose-induced PKC translocation and to examine the role of endogenously released ANG II in myocyte PKC signaling, primary cultures of adult rat ventricular myocytes were exposed to normal (5 mmol/l) or high (25 mmol/l) glucose for 12-24 h. Western blot analysis indicated that adult rat ventricular myocytes coexpress six PKC isozymes (alpha, beta(1,) beta(2,) delta, epsilon, and zeta). Translocation of five PKC isozymes (beta(1), beta(2), delta, epsilon, and zeta) was detected in response to 25 mmol/l glucose. Inhibition of phospholipase C with tricyclodecan-9-yl-xanthogenate blocked glucose-induced translocation of PKC-beta(2), -delta, and -zeta. Inhibition of tyrosine kinase with genistein blocked glucose-induced translocation of PKC-beta(1) and -delta, whereas chelation of intracellular Ca(2+) with 1,2-bis(2-aminophenoxy)ethane N,N,N,'N'-tetraacetic acid blocked translocation of PKC-beta(1) and -beta(2). Enzyme-linked immunosorbent assay performed on culture media from myocytes maintained in 25 mmol/l glucose detected a twofold increase in ANG II. Addition of an AT-1R antagonist (losartan; 100 nmol/l) to myocyte cultures blocked translocation of PKC-beta(1), -beta(2), -delta, and -epsilon. Phosphorylation of troponin (Tn) I was increased in myocytes exposed to 25 mmol/l glucose. Losartan selectively inhibited Tn I serine phosphorylation but did not affect phosphorylation at threonine residues. We concluded that 1) 25 mmol/l glucose triggers the release of ANG II by myocytes, resulting in activation of the ANG II autocrine pathway; 2) differential translocation of myocyte PKC isozymes occurs in response to 25 mmol/l glucose and ANG II; and 3) AT-1R-dependent PKC isozymes (beta(1), beta(2), delta, and epsilon) target Tn I serine residues. PMID- 11473057 TI - Effect of antioxidant treatment of streptozotocin-induced diabetic rats on endoneurial blood flow, motor nerve conduction velocity, and vascular reactivity of epineurial arterioles of the sciatic nerve. AB - We have shown that diabetes-induced reduction in endoneurial blood flow (EBF) and impaired endothelium-dependent vascular relaxation precede slowing of motor nerve conduction velocity (MNCV) and decreased sciatic nerve Na(+)/K(+) ATPase activity. Furthermore, vascular dysfunction was accompanied by an accumulation of superoxide in arterioles that provide circulation to the sciatic nerve. In the present study, we examined the effect that treatment of streptozotocin-induced diabetic rats with antioxidants has on vascular and neural function. Diabetic rats were treated with 0.5% alpha-lipoic acid as a diet supplement or with hydroxyethyl starch deferoxamine (HES-DFO) by weekly intravenous injections at a dose of 75 mg/kg. The treatments significantly improved diabetes-induced decrease in EBF, acetylcholine-mediated vascular relaxation in arterioles that provide circulation to the region of the sciatic nerve, and MNCV. The treatments also reduced the production of superoxide by the aorta and superoxide and peroxynitrite by arterioles that provide circulation to the region of the sciatic nerve. Treating diabetic rats with alpha-lipoic acid prevented the diabetes induced increase in thiobarbituric acid-reactive substances in serum and significantly improved lens glutathione levels. In contrast, treating diabetic rats with HES-DFO did not prevent diabetes-induced changes of either of these markers of oxidative stress. Diabetes-induced increase in sciatic nerve conjugated diene levels was not improved by treatment with either alpha-lipoic acid or HES-DFO. Treating diabetic rats with alpha-lipoic acid but not HES-DFO partially improved sciatic nerve Na(+)/K(+) ATPase activity and myo-inositol content. The increase in sciatic nerve sorbitol levels in diabetic rats was unchanged by either treatment. These studies suggest that diabetes-induced oxidative stress and the generation of superoxide may be partially responsible for the development of diabetic vascular and neural complications. PMID- 11473058 TI - Abnormalities of retinal metabolism in diabetes and experimental galactosemia. VII. Effect of long-term administration of antioxidants on the development of retinopathy. AB - Antioxidants were administered to diabetic rats and experimentally galactosemic rats to evaluate the ability of these agents to inhibit the development of diabetic retinopathy. Alloxan diabetic rats and nondiabetic rats that were fed 30% galactose randomly received standard diets or the diets supplemented with ascorbic acid and alpha-tocopherol (vitamins C+E diet) or a more comprehensive mixture of antioxidants (multi-antioxidant diet), including Trolox, alpha tocopherol, N-acetyl cysteine, ascorbic acid, beta-carotene, and selenium. Diabetes or galactose feeding of at least 12 months resulted in pericyte loss, acellular capillaries, and basement membrane thickening. Compared with diabetic controls, the development of acellular capillaries was inhibited by 50% (P < 0.05) in diabetic rats that received supplemental vitamins C+E, and the number of pericyte ghosts tended to be reduced. The vitamins C+E supplement had no beneficial effect in galactosemic rats, but these rats consumed only approximately half as much of the antioxidants as the diabetic rats. The multi antioxidant diet significantly inhibited ( approximately 55-65%) formation of both pericyte ghosts and acellular capillaries in diabetic rats and galactosemic rats (P < 0.05 vs. controls), without affecting the severity of hyperglycemia. Parameters of retinal oxidative stress, protein kinase C activity, and nitric oxides remained elevated for at least 1 year of hyperglycemia, and these abnormalities were normalized by multi-antioxidant therapy. Thus, long-term administration of antioxidants can inhibit the development of the early stages of diabetic retinopathy, and the mechanism by which this action occurs warrants further investigation. Supplementation with antioxidants can offer an achievable and inexpensive adjunct therapy to help inhibit the development of retinopathy in diabetes. PMID- 11473059 TI - Susceptibility and negative epistatic loci contributing to type 2 diabetes and related phenotypes in a KK/Ta mouse model. AB - The KK/Ta mouse strain serves as a suitable polygenic model for human type 2 diabetes. Using 93 microsatellite markers in 208 KK/Ta x (BALB/c x KK/Ta)F1 male backcross mice, we carried out a genome-wide linkage analysis of KK/Ta alleles contributing to type 2 diabetes and related phenotypes, such as obesity and dyslipidemia. We identified three major chromosomal intervals significantly contributing to impaired glucose metabolism: one quantitative trait locus for impaired glucose tolerance on chromosome 6 and two loci for fasting blood glucose levels on chromosomes 12 and 15. The latter two loci appeared to act in a complementary fashion. Two intervals showed significant linkages for serum triglyceride levels, one on chromosome 4 and the other on chromosome 8. The KK allele on chromosome 8 acts to promote serum triglyceride levels, whereas the KK allele on chromosome 4 acts to suppress this effect in a recessive fashion. In addition, it is suggested that the chromosome 4 locus also acts to downregulate body weight and that the chromosome 8 locus acts to upregulate serum insulin levels. Our data clearly showed that each disease phenotype of type 2 diabetes and related disorders in KK/Ta mice is under the control of separate genetic mechanisms. However, there appear to be common genes contributing to different disease phenotypes. There are potentially important candidate genes that may be relevant to the disease. PMID- 11473060 TI - New amino acid substitutions in the IRS-2 gene in Finnish and Chinese subjects with late-onset type 2 diabetes. AB - We investigated the significance of the variants of the IRS-2 gene in patients with type 2 diabetes. The entire coding part of the IRS-2 gene was screened by single-strand conformation polymorphism analysis in 40 Chinese and 40 Finnish patients with late-onset type 2 diabetes. The association of the variants of the IRS-2 gene with type 2 diabetes was studied in 85 Finnish diabetic patients and 82 Finnish control subjects and in 100 Chinese diabetic patients and 85 Chinese control subjects. The four variants predicting structural changes in the insulin receptor substrate (IRS)-2 protein included an insertion of AAC (Asn) in the Asn repeat sequence centered around codons 29-36 (allele frequencies of 0 vs. 0.6% and 1.5 vs. 0%), the Ala157Thr substitution (0 vs. 0% and 0.5 vs. 0%), the Leu647Val substitution (0.6 vs. 0% and 0 vs. 0%), and the Gly1057Asp polymorphism (31 vs. 31% and 35 vs. 30%) (P = NS for all comparisons). Furthermore, six silent variants were observed (CGC147CGG, CCC155CCG, GCC156GCT, AGT723AGC, TGT816TGC, and CCC829CCT). The Gly1057Asp polymorphism was not associated with insulin resistance or impaired insulin secretion in Finnish subjects with normal glucose tolerance (n = 295) or impaired glucose tolerance (n = 38). These data indicate that structural variants of the IRS-2 gene were uncommon in Finnish and Chinese patients with type 2 diabetes. Thus, the variants in the coding part of the IRS-2 gene are unlikely to have a major role in the development of type 2 diabetes in Finnish or Chinese subjects. PMID- 11473061 TI - A cluster of three single nucleotide polymorphisms in the 3'-untranslated region of human glycoprotein PC-1 gene stabilizes PC-1 mRNA and is associated with increased PC-1 protein content and insulin resistance-related abnormalities. AB - Glycoprotein PC-1 inhibits insulin signaling and, when overexpressed, plays a role in human insulin resistance. Mechanisms of PC-1 overexpression are unknown. We have identified a haplotype in the 3'-untranslated region of the PC-1 gene that may modulate PC-1 expression and confer an increased risk for insulin resistance. Individuals from Sicily, Italy, carrying the "P" haplotype (i.e., a cluster of three single nucleotide polymorphisms: G2897A, G2906C, and C2948T) were at higher risk (P < 0.01) for insulin resistance and had higher (P < 0.05) levels of plasma glucose and insulin during an oral glucose tolerance test and higher levels of cholesterol, HDL cholesterol, and systolic blood pressure. They also had higher (P < 0.05-0.01) PC-1 protein content in both skeletal muscle and cultured skin fibroblasts. In CHO cells transfected with either P or wild-type cDNA, specific PC-1 mRNA half-life was increased for those transfected with P (t/2 = 3.73 +/- 1.0 vs. 1.57 +/- 0.2 h; P < 0.01). In a population of different ethnicity (Gargano, East Coast Italy), patients with type 2 diabetes (the most likely clinical outcome of insulin resistance) had a higher P haplotype frequency than healthy control subjects (7.8 vs. 1.5%, P < 0.01), thus replicating the association between the P allele and the insulin resistance-related abnormalities observed among Sicilians. In conclusion, we have identified a possible molecular mechanism for PC-1 overexpression that confers an increased risk for insulin resistance-related abnormalities. PMID- 11473062 TI - What is so bad about a big baby? PMID- 11473063 TI - Third-trimester maternal glucose levels from diurnal profiles in nondiabetic pregnancies: correlation with sonographic parameters of fetal growth. AB - OBJECTIVE: To assess the 24-h glucose levels in a group of nondiabetic, nonobese pregnant women and to verify the presence of correlations between maternal glucose levels and sonographic parameters of fetal growth. RESEARCH DESIGN AND METHODS: A total of 66 Caucasian nonobese pregnant women with normal glucose challenge tests (GCT) enrolled in the study; from this population, we selected 51 women who delivered term (from 37 to 42 weeks completed) live-born infants without evidence of congenital malformations. The women were requested to have three main meals and to perform daily glucose profiles fortnightly from 28-38 weeks without modifying their lifestyle or following any dietary restriction. All subjects were taught how to monitor their blood glucose by using a reflectance meter. Fetal biometry was evaluated by ultrasound scan according to standard methodology at 22, 28, 32, and 36 weeks of pregnancy. RESULTS: The overall daily mean glucose level during the third trimester was 74.7 +/- 5.2 mg/dl. Daily mean glucose values increased between 28 (71.9 +/- 5.7 mg/dl) and 38 (78.3 +/- 5.4 mg/dl) weeks of pregnancy. We found a significant positive correlation at 28 weeks between 1-h postprandial glucose values and fetal abdominal circumference (AC). At 32 weeks, we documented positive correlations between fetal AC and maternal blood glucose levels 1 h after breakfast, 1 and 2 h after lunch, and 1 and 2 h after dinner. At 36 weeks, there was a positive correlation between fetal AC and 1- and 2-h postprandial blood glucose levels. In addition, there was a negative correlation between head-abdominal circumference ratio and 1-h postprandial blood glucose values. CONCLUSIONS: This longitudinal study first provides a contribution toward the definition of normoglycemia in nondiabetic, nonobese pregnant women; moreover, it reveals significant correlations of postprandial blood glucose levels with the growth of insulin-sensitive fetal tissues and, in particular, between 1-h postprandial blood glucose values and fetal AC. PMID- 11473064 TI - Serum type IV collagen in diabetic patients at risk for nephropathy. AB - OBJECTIVE: Whereas increased urinary excretion of type IV collagen, which is believed to reflect renal overproduction of this extracellular matrix protein in early diabetic nephropathy, has been confirmed in several studies, examination of serum concentrations of this analyte has yielded conflicting results. We sought to clarify the relationship between early renal dysfunction in diabetes and circulating type IV collagen concentrations. RESEARCH DESIGN AND METHODS: We measured serum (human) collagen IV concentrations by immunoassay in 109 patients with type 1 or type 2 diabetes and various amounts of albuminuria extending from the normo- to the macroalbuminuric range, and we examined its relationship to albumin excretion and to serum creatinine levels. RESULTS: Serum collagen IV concentrations (mean +/- SEM) were not significantly different in normoalbuminuric (219 +/- 10 ng/ml), microalbuminuric (209 +/- 6 ng/ml), or macroalbuminuric (206 +/- 7 ng/ml) diabetic subjects or in nondiabetic normal volunteers (206 +/- 10 ng/ml). Collagen IV concentrations showed no significant correlation (P > 0.25) with albumin excretion (r = -0.001), HbA(1c) (r = 0.030), or serum creatinine (r = -0.161) and were unrelated to urinary excretion of collagen IV in the subset of subjects in whom these data were available. CONCLUSIONS: The results of this cross-sectional analysis discount the utility of measurement of the serum concentration of collagen IV as an indicator of early renal dysfunction in diabetes. Increased urine excretion of collagen IV without a significant change in the serum concentration is consistent with a renal origin of this analyte in early diabetic nephropathy. PMID- 11473065 TI - The diabetes network internet-based physical activity intervention: a randomized pilot study. AB - OBJECTIVE: Because of other competing priorities, physical activity (PA) is seldom addressed in a consistent way in either primary care or diabetes education. This 8-week pilot study evaluated the short-term benefits of an Internet-based supplement to usual care that focused on providing support for sedentary patients with type 2 diabetes to increase their PA levels. RESEARCH DESIGN AND METHODS: A total of 78 type 2 diabetic patients (53% female, average age 52.3 years) were randomized to the Diabetes Network (D-Net) Active Lives PA Intervention or an Internet information-only condition. The intervention condition received goal-setting and personalized feedback, identified and developed strategies to overcome barriers, received and could post messages to an on-line "personal coach," and were invited to participate in peer group support areas. Key outcomes included minutes of PA per week and depressive symptomatology. RESULTS: There was an overall moderate improvement in PA levels within both intervention and control conditions, but there was no significant improvement in regard to condition effects. There was substantial variability in both site use and outcomes within the intervention and control conditions. Internal analyses revealed that among intervention participants, those who used the site more regularly derived significantly greater benefits, whereas those in the control condition derived no similar benefits with increased program use. CONCLUSIONS: Internet-based self-management interventions for PA and other regimen areas have great potential to enhance the care of diabetes and other chronic conditions. We conclude that greater attention should be focused on methods to sustain involvement with Internet-based intervention health promotion programs over time. PMID- 11473066 TI - The effect of aggressive versus standard lipid lowering by atorvastatin on diabetic dyslipidemia: the DALI study: a double-blind, randomized, placebo controlled trial in patients with type 2 diabetes and diabetic dyslipidemia. AB - OBJECTIVE: In patients with type 2 diabetes, intensive glucose regulation, although effective for microangiopathy, has not been shown to have unambiguous preventive effects on the occurrence of cardiovascular disease. Patients with diabetes show a characteristic dyslipidemia (high triglyceride level, low HDL cholesterol level). Aggressive reduction of triglycerides might be an effective method to reduce the cardiovascular risk in these patients. RESEARCH DESIGN AND METHODS: A double-blind, placebo-controlled, randomized study to assess the effect of 30 weeks of administration of atorvastatin 10 and 80 mg on plasma triglyceride levels in 217 patients with type 2 diabetes and fasting triglyceride levels between 1.5 and 6.0 mmol/l. RESULTS: Administration of atorvastatin 10 and 80 mg resulted in significant reductions (25 and 35%, respectively) of plasma triglyceride levels (both P < 0.001). The difference between 10 and 80 mg was not statistically significant (P > 0.5). Atorvastatin 10 mg provided significant reductions from baseline in total cholesterol (-30%, P < 0.001), LDL cholesterol (-40%, P < 0.001), and apolipoprotein B (-31%, P < 0.001), and significantly increased HDL cholesterol from baseline by 6% (P < 0.005). Atorvastatin 80 mg had a similar effect on HDL cholesterol (+5.2%, P < 0.005) but significantly decreased total cholesterol (-40%, P < 0.001), LDL cholesterol (-52%, P < 0.001), and apolipoprotein B (-40%, P < 0.001) more than atorvastatin 10 mg (P < 0.005). The side effects of atorvastatin 10 and 80 mg were similar and did not differ from the patients receiving placebo. CONCLUSIONS: Administration of 10- and 80-mg doses of atorvastatin provides similar, significant reductions from baseline in triglyceride levels in patients with type 2 diabetes. A higher dose of atorvastatin improves cholesterol-related parameters. Both doses were well tolerated in this patient population. PMID- 11473067 TI - Persistent differences among centers over 3 years in glycemic control and hypoglycemia in a study of 3,805 children and adolescents with type 1 diabetes from the Hvidore Study Group. AB - OBJECTIVE: Twenty-one international pediatric diabetes centers from 17 countries investigated the effect of simple feedback about the grand mean HbA(1c) level of all centers and the average value of each center on changes in metabolic control, rate of severe hypoglycemia, and insulin therapy over a 3-year period. RESEARCH DESIGN AND METHODS: Clinical data collection and determination of HbA(1c) levels were conducted at a central location in 1995 (n = 2,780, age 0-18 years) and 1998 (n = 2,101, age 11-18 years). RESULTS: Striking differences in average HbA(1c) concentrations were found among centers; these differences remained after adjustment for the significant confounders of sex, age, and diabetes duration. They were apparent even in patients with short diabetes duration and remained stable 3 years later (mean adjusted HbA(1c) level: 8.62 +/- 0.03 vs. 8.67 +/- 0.04 [1995 vs. 1998, respectively]). Three centers had improved significantly, four centers had deteriorated significantly in their overall adjusted HbA(1c) levels, and 14 centers had not changed in glycemic control. During the observation period, there were increases in the adjusted insulin dose by 0.076 U/kg, the adjusted number of injections by 0.23 injections per day, and the adjusted BMI by 0.95 kg/m(2). The 1995 versus 1998 difference in glycemic control for the seven centers could not be explained by prevailing insulin regimens or rates of hypoglycemia. CONCLUSIONS: This study reveals significant outcome differences among large international pediatric diabetes centers. Feedback and comparison of HbA(1c) levels led to an intensification of insulin therapy in most centers, but improved glycemic control in only a few. PMID- 11473068 TI - Long-term trends in childhood diabetes mortality: 1968-1998. AB - OBJECTIVE: In the context of recent improvements in type 1 diabetes therapy, to describe longitudinal trends in mortality attributable to childhood diabetes and to investigate socioeconomic and health services correlates of mortality. RESEARCH DESIGN AND METHODS: We extracted mortality data for 1968-1998 from National Center for Health Statistics files and covariates from the Bureau of Health Professions Area Resource File. Analytical techniques included linear and Poisson regression and standard descriptive statistics. RESULTS: Childhood (defined as 0-19 years of age) age-adjusted mortality from diabetes declined from 9.5 (1968) to 3.0 (1984) deaths per 10 million but remained relatively constant subsequently. All-cause childhood mortality, however, continued to decline. Older children experienced higher mortality rates, as did those living in counties with higher levels of unemployment. CONCLUSIONS: Despite recent improvements in therapy, diabetes-related mortality among children has not declined for 14 years. This finding may be partially attributable to sociodemographic factors influencing access to care, but the remaining mortality may defy available treatment methods. Reducing childhood diabetes mortality rates below the current apparent plateau may require new prevention and/or treatment strategies. PMID- 11473069 TI - The effect of day care exposure on the risk of developing type 1 diabetes: a meta analysis of case-control studies. AB - OBJECTIVE: Exposure to infections in infancy or childhood may be important in the pathogenesis of type 1 diabetes, but a protective role has also been suggested. We tested the hypothesis that increased early contact with infectious agents, measured by day care exposure, would decrease the risk of type 1 diabetes in childhood. RESEARCH DESIGN AND METHODS: We conducted a systematic review of case control studies. Meta-analysis was performed to combine results, assess for heterogeneity, and explore variation in study design. RESULTS: Several generally well-designed case-control studies show a statistically significant protective effect of day care on type 1 diabetes. However, meta-analysis revealed too much heterogeneity to accept the overall synthesis results and none of the studies used prerecorded data. Day care does seem to have a protective effect in the subgroup of children who will be diagnosed with type 1 diabetes before the age of 5 years (odds ratio = 0.6, 95% CI 0.5-0.8); however, this result is based on only two studies. CONCLUSIONS: Recall bias is one alternate explanation for these data; confirmation using prerecorded data is required. Such data could be prospectively measured in cohort studies of children at risk. We also suggest that information about day care attendance be measured in randomized trials of agents for the prevention of type 1 diabetes, as day care exposure could potentially modify the effect of the preventive agent. PMID- 11473070 TI - Evaluation of the insulin resistance syndrome in 5- to 10-year-old overweight/obese African-American children. AB - OBJECTIVE: To characterize the insulin sensitivity of overweight and obese 5- to 10-year-old (Tanner stage 1-3) African-American children screened for participation in a diabetes prevention study and to identify the association of insulin sensitivity with obesity, hyperlipidemia, and hypertension. RESEARCH DESIGN AND METHODS: Measures of insulin resistance (homeostasis model assessment) and insulin sensitivity (Matsuda and DeFronzo's whole-body insulin sensitivity) were calculated from a 2-h oral glucose tolerance test in 137 African-American children recruited into a diabetes prevention study. Measures of lipids (LDL, HDL, total cholesterol, and triglycerides), blood pressure, and body composition were obtained for a subset of the children. RESULTS: In response to a glucose challenge, girls and older and heavier children produced significantly more insulin. As BMI increased, there was a statistically significant decrease in insulin sensitivity, particularly in girls. Insulin sensitivity was inversely correlated with increases in blood pressure, triglycerides, subcutaneous fat, the percentage of total body fat, and Tanner stage, but it was not correlated with LDL and HDL. CONCLUSIONS: Reduced insulin sensitivity and the cluster of risk factors known as the insulin resistance syndrome (IRS) are already apparent in these overweight African-American children. Young African-American girls, in particular, already show evidence of hyperinsulinemia in response to a glucose load, suggesting that the early stages of metabolic decompensation that lead to type 2 diabetes are already occurring. Monitoring of those risk factors known to be part of IRS should become part of routine medical care for overweight or obese African-American children. PMID- 11473071 TI - Long-term effectiveness of a quality improvement program for patients with type 2 diabetes in general practice. AB - OBJECTIVE: To assess the long-term effectiveness of a quality improvement program on care provided and patient outcomes in patients with diabetes. RESEARCH DESIGN AND METHODS: A nonrandomized trial was performed with 312 patients with type 2 diabetes in the intervention group and 77 patients with type 2 diabetes in the reference group. The follow-up period was 42 months. The quality improvement program focused on improving both the provision of diabetes care and the patient outcomes. The program consisted of clinical practice guidelines, postgraduate education, audit and feedback, templates to register diabetes care, and a recall system. Data on the care provided were abstracted from medical records. Main outcomes on the provision of care were annual number of patient visits, blood pressure, and HbA(1c) and blood lipid levels. Main patient outcomes were blood pressure and HbA(1c) and blood lipid levels. Multilevel analysis was used to adjust for dependency between repeated observations within one patient and for clustering of patients within general practices. RESULTS: Patients in the intervention group received care far more in accordance with the guidelines than patients in the reference group. Odds ratios ranged from 2.43 (95% CI 1.01-5.82) for the measurement of urine albumin to 12.08 (4.70-31.01) for the measurement of blood pressure. No beneficial effect was found on any patient outcome. CONCLUSIONS: The quality improvement program improved the provision of diabetes care, but this was not accompanied by any effect on patient outcomes. PMID- 11473072 TI - Windows of opportunity to improve diabetes care when patients with diabetes are hospitalized for other conditions. AB - OBJECTIVE: The overwhelming majority of hospitalizations for patients with diabetes occur for treatment of comorbid conditions. This study assessed broad based interventions to improve diabetes care for patients hospitalized with cardiac conditions. RESEARCH DESIGN AND METHODS: A pre-post quasi-experimental study design was used to evaluate the implementation of two quality improvement interventions: 1) revision of the hospital's capillary blood glucose monitoring form into a color-coded process control chart and 2) a clinical path for type 2 diabetes as a secondary diagnosis. Interventions were implemented on the medical and surgical cardiac care units (not including the intensive care units on these services) of a tertiary academic medical center. A chart abstraction sample included 328 subjects with no exposure to the interventions and 336 subjects hospitalized after both interventions were implemented. Telephone surveys were conducted after discharge on 446 patients. RESULTS: The frequency of patients with severe hyperglycemia (at least one glucose level >400 mg/dl) and prolonged hyperglycemia (at least three consecutive glucose levels >250 mg/dl) decreased from 12 and 17% preintervention to 6.6 and 10% postintervention (P = 0.017 and P = 0.013, respectively). We found that 9% of the patients preintervention and 5% of the patients postintervention (P = 0.05) had nosocomial infections. Patient reported receipt of self-care instruction varied from 44 to 69% on nine survey items preintervention. Postintervention linear regression slopes for receipt of self-care instruction were all greater than preintervention slopes, but the differences did not achieve statistical significance. We found that 40% of the patients had important diabetes knowledge deficits. CONCLUSIONS: Our broad-based interventions were associated with a decreased frequency of prolonged and severe hyperglycemia and a decreased frequency of nosocomial infections. We also identified opportunities to improve diabetes self-care instruction before discharge and to address important knowledge deficits of patients. PMID- 11473073 TI - High prevalence of type 2 diabetes in all ethnic groups, including Europeans, in a British inner city: relative poverty, history, inactivity, or 21st century Europe? AB - OBJECTIVE: To compare the prevalence of type 2 diabetes in white Europeans and individuals of African-Caribbean and Pakistani descent. RESEARCH DESIGN AND METHODS: Random sampling of population-based registers in inner-city Manchester, Britain's third most impoverished area. A total of 1,318 people (25-79 years of age) were screened (minimum response 67%); 533 individuals without known diabetes underwent 2-h glucose tolerance testing, classified by 1999 World Health Organization criteria. RESULTS: More than 60% of individuals reported household annual income < pound10,000 ($15,000) per year. Energetic physical activity was rare and obesity was common. Age-standardized (35-79 years) prevalence (mean 95% CI) of known and newly detected diabetes was 20% (17-24%) in Europeans, 22% (18 26%) in African-Caribbeans, and 33% (25-41%) in Pakistanis. Minimum prevalence (assuming all individuals not tested were normoglycemic) was 11% (8-14%), 19% (15 23%), and 32% (24-40%), respectively. Marked changes in prevalence represent only small shifts in glucose distributions. Regression models showed that greater waist girth, lower height, and older age were independently related to plasma glucose levels, as was physical activity. Substituting BMI and waist-to-hip ratio revealed their powerful contribution. CONCLUSIONS: A surprisingly high prevalence of diabetes, despite expected increases with new lower criteria, was found in Europeans, as previously established in Caribbeans and Pakistanis. Lower height eliminated ethnic differences in regression models. History and relative poverty, which cosegregate with obesity and physical inactivity, are likely contributors. Whatever the causes, the implications for health services are alarming, although substantial preventive opportunities through small reversals of glucose distributions are the challenge. PMID- 11473074 TI - The marital relationship and psychosocial adaptation and glycemic control of individuals with diabetes. AB - OBJECTIVE: To explore the relationship between marital relationship domains (i.e., intimacy and adjustment) and glycemic control and psychosocial adaptation to diabetes. RESEARCH DESIGN AND METHODS: A total of 78 insulin-treated adults with both type 1 and type 2 diabetes were assessed on a single occasion. They completed two marital quality measures (Spanier Dyadic Adjustment Scale and Personal Assessment of Intimacy in Relationships Scale) and four quality-of-life measures (Diabetes Quality of Life Scale, Medical Outcomes Study Health Survey, Problem Areas in Diabetes Scale, and Positive and Negative Affect Scale). Glycemic control was assessed by HbA(1c). Demographic data (age, sex, type and duration of diabetes, years married, other medical conditions, family history, disability, and years of education) were gathered from the chart and questionnaires. RESULTS: Concerning psychosocial adaptation, both of the marital quality measures were predictors of aspects of adaptation. Better marital satisfaction was related to higher levels of diabetes-related satisfaction and less impact, as well as less diabetes-related distress and better general quality of life. Higher levels of marital intimacy were related to better diabetes specific and general quality of life. Concerning glycemic control, there was a nonsignificant trend for marital adjustment scores to relate to HbA(1c) (P = 0.0568). CONCLUSIONS: For insulin-treated adults with diabetes, quality of marriage is associated with adaptation to diabetes and other aspects of health related quality of life. The suggestive finding that marital adjustment may relate to glycemic control warrants further study. Future work should also explore the impact of couples-focused interventions on adaptation, adherence, and glycemic control. PMID- 11473075 TI - Is diabetes treated as an acute or chronic illness in community family practice? AB - OBJECTIVE: Poor quality of diabetes care has been ascribed to the acute care focus of primary care practice. A better understanding of how time is spent during outpatient visits for diabetes compared with visits for acute conditions and other chronic diseases may facilitate the design of programs to enhance diabetes care. RESEARCH DESIGN AND METHODS: Research nurses directly observed consecutive outpatient visits during two separate days in 138 community family physician offices. Time use was categorized into 20 different behaviors using the Davis Observation Code (DOC). Time use was compared for visits for diabetes, other chronic conditions, and acute illnesses during 1,867 visits by patients > or =40 years of age. RESULTS: Of 20 DOC behavioral categories, 10 exhibited differences among the three groups. Discriminant analysis identified two distinct factors that distinguished visits for chronic disease from visits for acute illness and visits for diabetes from those for other chronic diseases. Compared with visits for other chronic diseases, visits for diabetes devoted a greater proportion of time to nutrition counseling, health education, and feedback on results and less time to chatting. Compared with visits for acute illness, visits for diabetes were longer and involved a higher proportion of dietary advice, negotiation, and assessment of compliance. CONCLUSIONS: Visits for diabetes are distinct from visits for other chronic diseases and acute illnesses in ways that may facilitate patient self-management. Novel quality-improvement interventions could support and expand existing differences between family physicians' current approaches to care of diabetes and other chronic and acute illnesses. PMID- 11473076 TI - Postchallenge hyperglycemia and mortality in a national sample of U.S. adults. AB - OBJECTIVE: Although postchallenge hyperglycemia is a well-established feature of type 2 diabetes, its association with risk of mortality is uncertain. Therefore, the aim of this study was to assess the independent association of fasting and 2 h glucose levels with all-cause and cardiovascular disease (CVD) mortality. RESEARCH DESIGN AND METHODS: We analyzed data from the Second National Health and Nutrition Examination Survey (NHANES II) Mortality Study, a prospective cohort study of U.S. adults examined in the NHANES II, and focused on the 3,092 adults aged 30-74 years who underwent an oral glucose tolerance test at baseline (1976 1980). Deaths were identified from U.S. national mortality files from 1976 to 1992. To account for the complex survey design, we used SUDAAN statistical software for weighted analysis. RESULTS: Compared with their normoglycemic counterparts (fasting glucose [FG] < 7.0 and 2-h glucose < 7.8 mmol/l), adults with fasting and postchallenge hyperglycemia (FG > or =7.0 and 2-h glucose > or =11.1 mmol/l) had a twofold higher risk of death after 16 years of follow-up (age and sex-adjusted relative hazard [RH] 2.1, 95% CI 1.4-3.2). However, adults with isolated postchallenge hyperglycemia (FG < 7.0 and 2-h glucose > or =11.1 mmol/l) were also at higher risk of death (1.6, 1.0-2.6). In proportional hazards analysis, FG (fully adjusted RH 1.10 per 1 SD; 95% CI 1.01, 1.22) and 2-h glucose (1.14, 1.00-1.29) showed nearly identical predictive value for mortality. Similar trends were observed for CVD mortality. CONCLUSIONS: These results suggest that postchallenge hyperglycemia is associated with increased risk of all-cause and CVD mortality independently of other CVD risk factors. PMID- 11473077 TI - Fasting plasma homocysteine levels in the insulin resistance syndrome: the Framingham offspring study. AB - OBJECTIVE: Insulin resistance, associated metabolic abnormalities, and elevated homocysteine levels are risk factors for cardiovascular disease (CVD). We examined relationships between homocysteine levels and features of insulin resistance syndrome (IRS). RESEARCH DESIGN AND METHODS: We measured clinical characteristics, plasma levels of fasting homocysteine, folate, B vitamins, creatinine, and fasting and 2-h insulin and glucose levels after a 75-g oral glucose tolerance test in 2,214 subjects without CVD at the fifth examination (1991-1995) of the Framingham Offspring Study. After excluding 203 subjects with diabetes, the remaining 2,011 subjects were categorized as having none, one, two, or all three of the phenotypes of IRS: impaired glucose tolerance, hypertension, and/or a central metabolic syndrome (two or more traits: obesity, dyslipidemia, or hyperinsulinemia). In addition, in 1,592 subjects attending the sixth examination (1995-1998), we measured the urine albumin/creatinine ratio (UACR). Age-, sex-, creatinine-, vitamin-, and UACR-adjusted mean homocysteine levels or proportions with homocysteine >14 micromol/l in each phenotypic category and differences between categories were assessed with regression models. RESULTS: The mean age of the subjects was 54 years (range 28-82); 55% were women, 12.3% had hyperinsulinemia, and 15.9% had two or more of the IRS phenotypes. Adjusted mean homocysteine levels were higher comparing those with hyperinsulinemia (9.8 micromol/l) and those without (9.4 micromol/l, P = 0.04) and were higher among subjects with two or more IRS phenotypes (9.9 micromol/l) compared with those with 1 or no phenotype (9.3 micromol/l, P = 0.003). Mean UACR levels were also higher among subjects with two or more IRS phenotypes (7.2 mg/g) compared with those with 1 or no phenotype (5.5 mg/g, P = 0.007). CONCLUSIONS: Hyperhomocysteinemia and abnormal urinary albumin excretion are both associated with hyperinsulinemia and may partially account for increased risk of CVD associated with insulin resistance. Because hyperhomocysteinemia and microalbuminuria also reflect endothelial injury, these observations also support the hypothesis that endothelial dysfunction is associated with expression of the IRS. PMID- 11473078 TI - Utilization of oral hypoglycemic agents in a drug-insured U.S. population. AB - OBJECTIVE: Clinical trials provide information regarding the safety and efficacy of medications used to manage type 2 diabetes but do not elucidate drug effectiveness in a typical managed care environment. The aim of this study was to characterize "real-world" drug utilization patterns from both a prescriber and a patient perspective. RESEARCH DESIGN AND METHODS: We conducted a retrospective analysis of a large administrative pharmacy claims database, using data on continuously pharmacy benefit-eligible members prescribed oral hypoglycemic agents (OHAs). RESULTS: The 12-month persistence rate for the OHA cohort was low, ranging from 31% for alpha-glucosidase inhibitors to 60% for metformin; compliance rates varied between 70 and 80%. During the first 12 months of therapy, 36% of the patients remaining on therapy at 12 months had one or more therapy modifications. The mean number of therapy changes increased with the length of patient follow-up, with more than half of all patients experiencing at least one therapy change over the duration of follow-up. CONCLUSIONS: These findings document the wide variation in utilization patterns associated with pharmacological management of type 2 diabetes, suggesting that opportunity exists to optimize its pharmacological management. PMID- 11473079 TI - Glucagon-like peptide-1 infusion must be maintained for 24 h/day to obtain acceptable glycemia in type 2 diabetic patients who are poorly controlled on sulphonylurea treatment. AB - OBJECTIVE: To assess the efficacy and safety of glucagon-like peptide-1 (GLP-1) on the plasma glucose level when given as a continuous infusion for either 16 or 24 h per day to type 2 diabetic patients who were poorly controlled on sulfonylurea treatment. RESEARCH DESIGN AND METHODS: This single-center, randomized, parallel, double-blind, placebo-controlled trial was conducted in 40 hospitalized patients who were randomized to receive infusions of either placebo or GLP-1 4 or 8 ng. kg(-1). min(-1) for either 16 or 24 h per day for 7 days. At predetermined intervals, 24-h profiles of glucose, glucagon, and insulin were measured. Adverse events and clinical chemistry and hematology were recorded. RESULTS: For all active treatment groups, the change in average glucose (area under the curve [AUC] for day 7 minus AUC for day 0 divided by 24 h) was statistically significantly different from placebo (P < or = 0.001). The GLP-1 8 ng. kg(-1). min(-1) dose given for 24 h was more efficacious than any of the other doses (P < or = 0.05). Nocturnal and fasting plasma glucose levels at day 7 were greater in the 16-h groups compared with the 24-h groups (P < or = 0.05). Insulin AUC did not show any treatment effect for any of the treatment groups when change was assessed from day 0 to day 7. However, for the 16-h groups, the pattern of the insulin profiles changed; the insulin profiles were considerably higher during the initial 3-4 h after restart of the GLP-1 infusion on day 7, although there was a tendency for insulin levels to decrease during the afternoon and evening. Glucagon AUC decreased significantly for all active treatment groups compared with placebo. GLP-1 was generally well tolerated. CONCLUSIONS: This study demonstrated that GLP-1 should be given continuously to obtain the most optimal glycemic control. Because of the short plasma half-life of native GLP-1, long-acting derivatives should be developed to make GLP-1 treatment clinically relevant. PMID- 11473080 TI - Impact of diabetes on long-term survival after acute myocardial infarction: comparability of risk with prior myocardial infarction. AB - OBJECTIVE: To determine the effect of diabetes on long-term survival after acute myocardial infarction and to compare its effect with that of a previous myocardial infarction. RESEARCH DESIGN AND METHODS: In a prospective cohort study, we followed 1,935 patients hospitalized with a confirmed acute myocardial infarction at 45 U.S. medical centers between 1989 and 1993, as part of the Determinants of Myocardial Infarction Onset Study. Trained interviewers performed chart reviews and face-to-face interviews with all patients. We analyzed survival using Cox proportional hazards regression to control for potentially confounding factors. RESULTS: Of the 1,935 patients, 320 (17%) died during a mean follow-up of 3.7 years. A total of 399 patients (21%) had previously diagnosed diabetes. Diabetes was associated with markedly higher total mortality in unadjusted (hazard ratio [HR] 2.4; 95% CI 1.9-3.0) and adjusted (1.7; 1.3-2.1) analyses. The magnitude of the effect of diabetes was identical to that of a previous myocardial infarction. The effect of diabetes was not significantly modified by age, smoking, household income, use of thrombolytic therapy, type of hypoglycemic treatment, or duration of diabetes, but the risk associated with diabetes was higher among women than men (adjusted HRs 2.7 vs. 1.3, P = 0.01). CONCLUSIONS: Diabetes is associated with markedly increased mortality after acute myocardial infarction, particularly in women. The increase in risk is of the same magnitude as a previous myocardial infarction and provides further support for aggressive treatment of coronary risk factors among diabetic patients. PMID- 11473081 TI - Cardiovascular drug use and hospitalizations attributable to type 2 diabetes. AB - OBJECTIVE: To investigate cardiovascular drug use and hospitalizations attributable to type 2 diabetes from 1 year before until 6 years after the start of oral antidiabetic therapy. RESEARCH DESIGN AND METHODS: In this cohort study, 2,584 patients with type 2 diabetes were selected from the PHARMO Record Linkage System, comprising pharmacy records and hospitalizations for all 320,000 residents of six Dutch cities. Patients with type 2 diabetes were identified as incident oral antidiabetic drug users between 1992 and 1997. Nondiabetic subjects were 1:1-matched for age, sex, pharmacy, and index date and received no insulin, oral antidiabetic drugs, or glucose-testing supplies. RESULTS: Patients with type 2 diabetes were more likely to use cardiovascular drugs (RR 1.28 [95% CI 1.23 1.34]) and to be hospitalized because of cardiovascular diseases (1.54 [1.33 1.78]) after the start of oral antidiabetic therapy than nondiabetic subjects. Differences between patients with type 2 diabetes and nondiabetic subjects lessened from 1 year before until 6 years after the start of oral antidiabetic therapy, reflected by decreasing attributable risks for diuretics, beta-blockers, calcium channel blockers, and cardiac and antithrombotic drugs. The difference in use of angiotensin-converting enzyme inhibitors and lipid-lowering drugs increased. Cardiovascular hospitalizations attributable to type 2 diabetes were approximately 50% in the years close to the start of oral antidiabetic treatment and decreased to approximately 33% in the following years. CONCLUSIONS: Although cardiovascular drug use and hospitalizations remained increased in patients with type 2 diabetes after the start of oral antidiabetic therapy, cardiovascular drug use attributable to type 2 diabetes decreased after the start of oral antidiabetic therapy, especially beta-blockers, whereas cardiovascular hospitalizations first decreased and then stabilized. PMID- 11473082 TI - Peripheral arterial disease in diabetic and nondiabetic patients: a comparison of severity and outcome. AB - OBJECTIVE: The aim of this study was to quantify the distribution of peripheral arterial disease in the diabetic and nondiabetic population attending for angiography and to compare severity and outcome between both groups of patients. RESEARCH DESIGN AND METHODS: Randomly selected lower-extremity angiograms were examined according to the Bollinger system. Patient demographics and medical history were recorded and case notes were examined to determine which patients later underwent a revascularization procedure or amputation and which patients had died. RESULTS: A total of 136 arteriograms obtained between 1992 and 1996 were analyzed. The age (mean +/- SD) of the patients was 64.7 +/- 10.8 years. Diabetic patients (43%) and nondiabetic patients were of similar age (63.9 +/- 10.4 vs. 65.3 +/- 11.1 years, P = 0.43), with a similar history of smoking (81.0 vs. 76.9%, P = 0.26), ischemic heart disease (41.4 vs. 37.2%, P = 0.54), and hypercholesterolemia (24.4 vs. 30.8%, P = 0.48). However, there were a greater proportion of hypertensive patients in the diabetic group (63.8 vs. 39.7%, P = 0.006). Diabetic patients had greater severity of arterial disease in the profunda femoris and all arterial segments below the knee (P = 0.02). A greater number of amputations occurred in the diabetic group: diabetic patients were five times more likely to have an amputation (41.4 vs. 11.5%, odds ratio [OR] 5.4, P < 0.0001). Mortality was higher in the diabetic group (51.7 vs. 25.6%, OR 3.1, P = 0.002), and diabetic patients who died were younger at presentation than nondiabetic patients (64.7 +/- 11.4 vs. 71.1 +/- 8.7 years, P = 0.04). CONCLUSIONS: In patients with peripheral arterial disease, diabetic patients have worse arterial disease and a poorer outcome than nondiabetic patients. PMID- 11473083 TI - NeuroD/beta2 gene G-->A polymorphism may affect onset pattern of type 1 diabetes in Japanese. AB - OBJECTIVE: The majority of type 1 diabetes is considered to be autoimmune with, for the most part, abrupt development. However, type 1 diabetes with slow onset, or the so-called slowly progressive type 1 diabetes or latent autoimmune diabetes in adults, has been recently recognized and is considered to be autoimmune related. Although some investigators tried to explain the difference in onset pattern by the genetic background, including HLA type, it has not been established thus far. We hypothesized that the difference in onset pattern may relate to regeneration or differentiation of pancreatic beta-cells, and we therefore focused on the NeuroD/BETA2 gene, which encodes a transcription factor for the insulin gene and beta-cell differentiation. RESEARCH DESIGN AND METHODS: We examined the NeuroD/BETA2 gene polymorphism in 105 Japanese type 1 diabetic patients and in 122 nondiabetic Japanese subjects in a case-control study, and we stratified the patients according to their onset pattern and islet-associated autoantibody positivity. RESULTS: Regardless of the existence of islet-associated autoantibody, we found a significant difference in A allele frequency between type 1 diabetic patients with acute-onset type and control subjects. However, no difference was found between type 1 slow-onset diabetic patients and control subjects. CONCLUSIONS: These results support our hypothesis that NeuroD/BETA2 may affect the ability of regeneration of beta-cells, leading to a difference in the onset pattern and clinical course of type 1 diabetes. PMID- 11473084 TI - Effectiveness of the diabetic foot risk classification system of the International Working Group on the Diabetic Foot. AB - OBJECTIVE: To evaluate the effectiveness of a diabetic foot risk classification system by the International Working Group on the Diabetic Foot to predict clinical outcomes. RESEARCH DESIGN AND METHODS: A total of 225 diabetic patients were initially evaluated as part of a prospective case-control study at the University of Texas Health Science Center at San Antonio. Complete records were available for 213 patients for follow-up evaluation after 29 months. Upon enrollment, subjects were stratified into four risk groups based on the presence of risk factors according to the consensus of the International Working Group on the Diabetic Foot. Group 0 consisted of subjects without neuropathy, group 1 consisted of patients with neuropathy but without deformity or peripheral vascular disease (PVD), group 2 consisted of subjects with neuropathy and deformity or PVD, and group 3 consisted of patients with a history of foot ulceration or a lower-extremity amputation. RESULTS: Upon enrollment, patients in higher-risk groups had longer duration of diabetes, worse glycemic control, vascular and neuropathic variables, and more systemic complications of diabetes. During 3 years of follow-up, ulceration occurred in 5.1, 14.3, 18.8, and 55.8% of the patients in groups 0, 1, 2, and 3, respectively (linear-by-linear association, P < 0.001). All amputations were found in Groups 2 and 3 (3.1 and 20.9%, P < 0.001). CONCLUSIONS: The foot risk classification of the International Working Group on the Diabetic Foot predicts ulceration and amputation and can function as a tool to prevent lower-extremity complications of diabetes. PMID- 11473085 TI - Increased prevalence of impaired glucose tolerance in patients with painful sensory neuropathy. AB - OBJECTIVE: To characterize a cohort of patients with neuropathy and impaired glucose tolerance (IGT) but no other identifiable cause of neuropathy. Of patients with diabetes, 10% have peripheral neuropathy at the time of their diagnosis, suggesting that axonal injury may occur early in the course of glucose intolerance. The American Diabetes Association (ADA) revised diagnostic criteria to recognize IGT (a serum glucose between 140 and 200 mg/dl in a 2-h oral glucose tolerance test [OGTT]) as a risk factor for cardiovascular disease independent of development of diabetes. RESEARCH DESIGN AND METHODS: Using revised ADA criteria for diabetes and IGT, we prospectively evaluated 107 sequential patients with idiopathic neuropathy. RESULTS: A total of 13 of the 107 patients had diabetes, whereas 36 (34%) had IGT, nearly three times the prevalence in age-matched control subjects (P < 0.01). OGTT was often elevated, whereas both fasting plasma glucose and HbA(1c) were normal. Comparing patients with diabetes, IGT, or normal OGTT, age and BMI were similar. However, painful sensory symptoms were more common in patients with IGT and diabetes, and family history of neuropathy was significantly more common in normoglycemic patients. Electrodiagnostic findings of axonal injury were less severe in patients with IGT and were more likely to be confined to sensory fibers than in patients with diabetes. CONCLUSIONS: Our results suggest that IGT may cause or contribute to small-fiber neuropathy, which is similar in phenotype to the painful sensory neuropathy commonly encountered in diabetes. Two-hour OGTT is more sensitive than other measures of glucose handling in screening these patients. PMID- 11473086 TI - Parental history of diabetes modifies the association between abdominal adiposity and hyperglycemia. AB - OBJECTIVE: To examine whether the association between abdominal obesity and hyperglycemia differs according to the presence of a parental history of diabetes. RESEARCH DESIGN AND METHODS: We conducted a cross-sectional study of 3,068 men and women, aged 20-65 years, without known diabetes who were fasting participants of a population-based study in three Dutch towns. Hyperglycemia was defined as a fasting plasma glucose concentration of 6.1 mmol/l (American Diabetes Association criterion). Waist circumference was categorized according to previously defined waist action levels. All estimates were adjusted for age and town. RESULTS: The regression coefficients for the association between waist circumference and fasting plasma glucose were larger in participants who had a parental history of diabetes than in those who did not (men beta = 0.31 vs. 0.16 mmol/SD, P [for interaction] = 0.003; women beta = 0.24 vs. 0.11 mmol/SD, P = 0.002). Furthermore, larger waist circumference (men > or = 94 vs. < 94 cm, women > or = 88 vs. < 80 cm) was associated with a greater excess prevalence of hyperglycemia in participants who had a parental history of diabetes than in those who did not (men 12.4 vs. 2.0%, P = 0.03; women 13.6 vs. 5.9%, P = 0.05). Adjustment for physical activity, alcohol intake, smoking, and educational level did not materially change the results. CONCLUSIONS: These findings indicate that the association between abdominal obesity and hyperglycemia is stronger in the presence of a parental history of diabetes. Blood glucose screening may be warranted at lower levels of waist circumference in individuals with a parental history of diabetes. PMID- 11473087 TI - Autoimmune diabetes not requiring insulin at diagnosis (latent autoimmune diabetes of the adult): definition, characterization, and potential prevention. AB - Type 1 diabetes is caused by the immune-mediated destruction of islet insulin secreting beta-cells. This chronic destructive process is associated with both cellular and humoral immune changes in the peripheral blood that can be detected months or even years before the onset of clinical diabetes. Throughout this prediabetic period, metabolic changes, including altered glucose tolerance and reduced insulin secretion, deteriorate at variable rates and eventually result in clinical diabetes. A fraction of individuals with humoral immunological changes have clinical diabetes that initially is not insulin-requiring. The onset of diabetes in these patients is usually in adult life, and because their diabetes is at least initially not insulin-requiring, they appear clinically to be affected by type 2 diabetes. Such patients probably have the same disease process as patients with type 1 diabetes in that they have similar HLA genetic susceptibility as well as autoantibodies to islet antigens, low insulin secretion, and a higher rate of progression to insulin dependency. These patients are defined as being affected by an autoimmune type of diabetes not requiring insulin at diagnosis, which is also named latent autoimmune diabetes of the adult (LADA). Special attention should be paid to diagnose such patients because therapy may influence the speed of progression toward insulin dependency, and in this respect, efforts should be made to protect residual C-peptide secretion. LADA can serve as a model for designing new strategies for prevention of type 1 diabetes but also as a target group for prevention in its own right. PMID- 11473088 TI - Dermal neurovascular dysfunction in type 2 diabetes. AB - OBJECTIVE: To review evidence for a relationship between dermal neurovascular dysfunction and other components of the metabolic syndrome of type 2 diabetes. RESEARCH DESIGN AND METHODS: We review and present data supporting concepts relating dermal neurovascular function to prediabetes and the metabolic syndrome. Skin blood flow can be easily measured by laser Doppler techniques. RESULTS: Heat and gravity have been shown to have specific neural, nitrergic, and independent mediators to regulate skin blood flow. We describe data showing that this new tool identifies dermal neurovascular dysfunction in the majority of type 2 diabetic patients. The defect in skin vasodilation is detectable before the development of diabetes and is partially correctable with insulin sensitizers. This defect is associated with C-fiber dysfunction (i.e., the dermal neurovascular unit) and coexists with variables of the insulin resistance syndrome. The defect most likely results from an imbalance among the endogenous vasodilator compound nitric oxide, the vasodilator neuropeptides substance P and calcitonin gene-related peptide, and the vasoconstrictors angiotensin II and endothelin. Hypertension per se increases skin vasodilation and does not impair the responses to gravity, which is opposite to that of diabetes, suggesting that the effects of diabetes override and counteract those of hypertension. CONCLUSIONS: These observations suggest that dermal neurovascular function is largely regulated by peripheral C-fiber neurons and that dysregulation may be a component of the metabolic syndrome associated with type 2 diabetes. PMID- 11473089 TI - Platelet dysfunction in type 2 diabetes. AB - Insulin resistance is a uniform finding in type 2 diabetes, as are abnormalities in the microvascular and macrovascular circulations. These complications are associated with dysfunction of platelets and the neurovascular unit. Platelets are essential for hemostasis, and knowledge of their function is basic to understanding the pathophysiology of vascular disease in diabetes. Intact healthy vascular endothelium is central to the normal functioning of smooth muscle contractility as well as its normal interaction with platelets. What is not clear is the role of hyperglycemia in the functional and organic microvascular deficiencies and platelet hyperactivity in individuals with diabetes. The entire coagulation cascade is dysfunctional in diabetes. Increased levels of fibrinogen and plasminogen activator inhibitor 1 favor both thrombosis and defective dissolution of clots once formed. Platelets in type 2 diabetic individuals adhere to vascular endothelium and aggregate more readily than those in healthy people. Loss of sensitivity to the normal restraints exercised by prostacyclin (PGI(2)) and nitric oxide (NO) generated by the vascular endothelium presents as the major defect in platelet function. Insulin is a natural antagonist of platelet hyperactivity. It sensitizes the platelet to PGI(2) and enhances endothelial generation of PGI(2) and NO. Thus, the defects in insulin action in diabetes create a milieu of disordered platelet activity conducive to macrovascular and microvascular events. PMID- 11473090 TI - Complementary medicine: its hidden risks. PMID- 11473091 TI - The American Diabetes Association's 48th Annual Advanced Postgraduate Course. PMID- 11473092 TI - Plasma total homocysteine levels are associated with von Willebrand factor, soluble intercellular adhesion molecule-1, and soluble tumor necrosis factor alpha receptors in young type 1 diabetic patients without clinical evidence of macrovascular complications. PMID- 11473093 TI - High prevalence of hypovitaminosis D in female type 2 diabetic population. PMID- 11473094 TI - Absence of association of type 2 diabetes with CAPN10 and PC-1 polymorphisms in Oji-Cree. PMID- 11473095 TI - Internalized racism is associated with glucose intolerance among Black Americans in the U.S. Virgin Islands. PMID- 11473096 TI - Improvement in endothelial dysfunction with LDL cholesterol level < 80 mg/dl in type 2 diabetic patients. PMID- 11473097 TI - Maternal mortality in type 1 diabetes. PMID- 11473098 TI - Lack of compliance with home blood glucose monitoring predicts hospitalization in diabetes. PMID- 11473099 TI - Is there a predisposition to intestinal parasitosis in diabetic patients? PMID- 11473100 TI - Seasonal variation of glycemic control in type 2 diabetic patients. PMID- 11473101 TI - Serum nonesterified fatty acids are related with carotid atherosclerotic plaque in nonobese nonhypertensive Japanese type 2 diabetic patients. PMID- 11473102 TI - Increased prevalence of significant coronary artery calcification in patients with diabetes. PMID- 11473103 TI - Diabetes trends among American Indians and Alaska natives: 1990-1998. PMID- 11473104 TI - Increased prevalence of significant coronary artery calcification in patients with diabetes. PMID- 11473105 TI - Strong growth polarity of yeast prion fiber revealed by single fiber imaging. AB - Using the yeast prion as a model, we have developed a novel system to observe the growth of individual prion fibers directly. NM fragments, the prion-determining region of the yeast protein Sup35p, were labeled by either red or green fluorescent dyes, and the fiber growth was observed under a fluorescence microscope. When green-Sup35NM was added to the preformed fibers made of red Sup35NM, 70-97% of green fibers grew unidirectionally, from only one end of individual red fibers, whereas the remainder grew from both ends. Similarly, the majority of red fibers grew from only one end of green fibers when the order of addition was reversed. Sonication of preformed fibers to expose fresh ends did not change the results, excluding a possibility of occasional deformation of one end as the reason of the apparent unidirectional growth. These results indicate the polarity of Sup35 prion fibers and impose constraints on the models of fiber growth. PMID- 11473106 TI - The human estrogen receptor-alpha is a ubiquitinated protein whose stability is affected differentially by agonists, antagonists, and selective estrogen receptor modulators. AB - The human estrogen receptor alpha-isoform (ERalpha) is a nuclear transcription factor that displays a complex pharmacology. In addition to classical agonists and antagonists, the transcriptional activity of ERalpha can be regulated by selective estrogen receptor modulators, a new class of drugs whose relative agonist/antagonist activity is determined by cell context. It has been demonstrated that the binding of different ligands to ERalpha results in the formation of unique ERalpha-ligand conformations. These conformations have been shown to influence ERalpha-cofactor binding and, therefore, have a profound impact on ERalpha pharmacology. In this study, we demonstrate that the nature of the bound ligand also influences the stability of ERalpha, revealing an additional mechanism by which the pharmacological activity of a compound is determined. Of note we found that although all ERalpha-ligand complexes can be ubiquitinated and degraded by the 26 S proteasome in vivo, the mechanisms by which they are targeted for proteolysis appear to be different. Specifically, for agonist-activated ERalpha, an inverse relationship between transcriptional activity and receptor stability was observed. This relationship does not extend to selective estrogen receptor modulators and pure antagonists. Instead, it appears that with these compounds, the determinant of receptor stability is the ligand-induced conformation of ERalpha. We conclude that the different conformational states adopted by ERalpha in the presence of different ligands influence transcriptional activity directly by regulating cofactor binding and indirectly by modulating receptor stability. PMID- 11473107 TI - Histone deacetylase is a direct target of valproic acid, a potent anticonvulsant, mood stabilizer, and teratogen. AB - Valproic acid is widely used to treat epilepsy and bipolar disorder and is also a potent teratogen, but its mechanisms of action in any of these settings are unknown. We report that valproic acid activates Wntdependent gene expression, similar to lithium, the mainstay of therapy for bipolar disorder. Valproic acid, however, acts through a distinct pathway that involves direct inhibition of histone deacetylase (IC(50) for HDAC1 = 0.4 mm). At therapeutic levels, valproic acid mimics the histone deacetylase inhibitor trichostatin A, causing hyperacetylation of histones in cultured cells. Valproic acid, like trichostatin A, also activates transcription from diverse exogenous and endogenous promoters. Furthermore, valproic acid and trichostatin A have remarkably similar teratogenic effects in vertebrate embryos, while non-teratogenic analogues of valproic acid do not inhibit histone deacetylase and do not activate transcription. Based on these observations, we propose that inhibition of histone deacetylase provides a mechanism for valproic acid-induced birth defects and could also explain the efficacy of valproic acid in the treatment of bipolar disorder. PMID- 11473108 TI - Localization of the chaperone domain of FKBP52. AB - FKBP52, a multidomain peptidyl prolyl cis/trans-isomerase (PPIase), is found in complex with the chaperone Hsp90 and the co-chaperone p23. It displays both PPIase and chaperone activity in vitro. To localize these two activities to specific regions of the protein, we created and analyzed a set of fragments of FKBP52. The PPIase activity toward both peptides and proteins is confined entirely to domain 1 (amino acids 1-148). The chaperone activity, however, resides in the C-terminal part of FKBP52, mainly in the region between amino acids 264 and 400 (domain 3). Interestingly, this domain also contains the tetratricopeptide repeats, which are responsible for the binding to C-terminal amino acids of Hsp90. Competition assays with a C-terminal Hsp90 peptide suggest that the non-native protein and Hsp90 are bound by different regions within this domain. PMID- 11473109 TI - Reduced protein phosphatase 2A activity induces hyperphosphorylation and altered compartmentalization of tau in transgenic mice. AB - Hyperphosphorylated isoforms of the microtubule-associated protein tau are the major components of neurofibrillary lesions in Alzheimer's disease (AD). Protein phosphatase (PP) 2A is a major phosphatase implicated in tau dephosphorylation in vitro. Dephosphorylation of tau can be blocked in vivo by okadaic acid, a potent inhibitor of PP2A. Moreover, activity of PP2A is reduced in AD brains. To elucidate the role of PP2A in tau phosphorylation and pathogenesis, we expressed a dominant negative mutant form of the catalytic subunit Calpha of PP2A, L199P, in mice by using a neuron-specific promoter. We obtained mice with high expression levels of Calpha L199P in cortical, hippocampal, and cerebellar neurons. PP2A activity in brain homogenates of transgenic mice was reduced to 66%. Endogenous tau protein was hyperphosphorylated at distinct sites including the AT8 epitope Ser-202/Thr-205, a major AD-associated tau phosphoepitope. AT8 positive tau aggregates accumulated in the soma and dendrites of cortical pyramidal cells and cerebellar Purkinje cells and co-localized with ubiquitin. Our data establish that PP2A plays a crucial role in tau phosphorylation. Our results also show that reduced PP2A activity is associated with altered compartmentalization and ubiquitination of tau, resembling a key pathological finding in AD. PMID- 11473110 TI - Nucleotides provide a voltage-sensitive gate for the rapid anion channel of arabidopsis hypocotyl cells. AB - The rapid anion channel of Arabidopsis hypocotyl cells is highly voltage dependent. At hyperpolarized potentials, the channel is closed, and membrane depolarization is required for channel activation. We have previously shown that channel gating is regulated by intracellular nucleotides. In the present study, we further analyze the channel gating, and we propose a mechanism to explain its regulation by voltage. In the absence of intracellular nucleotides, closure at hyperpolarized voltages is abolished. Structure-function studies of adenyl nucleotides show that the apparent gating charge of the current increases with the negative charge carried by nucleotides. We propose that the fast anion channel is gated by the voltage-dependent entry of free nucleotides into the pore, leading to a voltage-dependent block at hyperpolarized potentials. In agreement with this mechanism in which intracellular nucleotides need to be recruited to the channel pore, kinetic analyses of whole-cell and single-channel currents show that the rate of closure is faster when intracellular nucleotide concentration is increased, whereas the rate of channel activation is unchanged. Furthermore, decreasing the concentration of extracellular chloride enhances the intracellular nucleotide block. This result supports the hypothesis of a mechanism in which blocking nucleotides and permeant anions interact within the channel pore. PMID- 11473111 TI - The heme environment of mouse neuroglobin. Evidence for the presence of two conformations of the heme pocket. AB - Neuroglobin (Ngb) is a newly discovered oxygen-binding heme protein that is primarily expressed in the brain of humans and other vertebrates. To characterize the structure/function relationships of this new heme protein, we have used resonance Raman spectroscopy to determine the structure of the heme environment in Ngb from mice. In the Fe(2+)CO complex, two conformations of the Fe-CO unit are present, one of which arises from an open conformation of the heme pocket in which the CO is not interacting with any nearby residue, and the other arises from a closed conformation where a positively charged residue near the CO group stabilizes the complex. For the Fe(2+)O(2) complex, we detect a single nu(Fe-OO) stretching mode at a frequency similar to that of oxymyoglobins and oxyhemoglobins of vertebrates (571 cm(-1)). Based on the Fe-C-O frequencies of the closed conformation of Ngb, a highly polar distal environment is indicated from which the O(2) off-rate is predicted to be lower than that of Mb. In the absence of exogenous ligands, a heme pocket residue coordinates to the heme iron, forming a six-coordinate complex, thereby predicting a low on-rate for exogenous ligands. These structural properties of the heme pocket of Ngb are discussed with respect to its proposed in vivo oxygen delivery function. PMID- 11473112 TI - delta Ef1 binds to a far upstream sequence of the mouse pro-alpha 1(I) collagen gene and represses its expression in osteoblasts. AB - The transcription of type I collagen genes is tightly regulated, but few cis acting elements have been identified that can modulate the levels of expression of these genes. Generation of transgenic mice harboring various segments of the mouse pro-alpha1(I) collagen promoter led us to suspect that a repressor element was located between -10.5 and -17 kilobase pairs. Stable and transient transfection experiments in ROS17/2.8 osteoblastic cells confirmed the existence of such a repressor element at about -14 kilobase pairs and showed that it consisted in an almost perfect three-time repeat of a 41-base pair sequence. This element, which we named COIN-1, contains three E2-boxes, and a point mutation in at least two of them completely abolished its repressor effect. In gel shift assays, COIN-1 bound a DNA-binding protein named delta EF1/ZEB-1, and mutations that abolished the repressor effect of COIN-1 also suppressed the binding of delta EF1. We also showed that the repressor effect of COIN-1 was not mediated by chromatin compaction. Furthermore, overexpression of delta EF1 in ROS17/2.8 osteoblastic cells enhanced the inhibitory effect of COIN-1 in a dose-dependent manner and repressed the expression of the pro-alpha 1(I) collagen gene. Thus, delta EF1 appears to repress the expression of the mouse pro-alpha 1(I) collagen gene, through its binding to COIN-1. PMID- 11473113 TI - Protein intermediate trapped by the simultaneous crystallization process. Crystal structure of an iron-saturated intermediate in the Fe3+ binding pathway of camel lactoferrin at 2.7 a resolution. AB - This is the first protein intermediate obtained in the crystalline state by the simultaneous process of Fe(3+) binding and crystal nucleation and is also the first structure of an intermediate of lactoferrin in the Fe(3+) binding pathway. Lactoferrin is an iron-binding 80-kDa glycoprotein. It binds Fe(3+) very tightly in a closed interdomain cleft in both lobes. The iron-free structure of lactoferrin, on the other hand, adopts an open conformation with domains moving widely apart. These studies imply that initial Fe(3+) binding must be in the open form. The protein intermediate was crystallized by the microdialysis method. The protein solution, with a concentration of 100 mg/ml in 10 mm Tris-HCl, pH 8.0, was loaded in a capillary and dialyzed against the same buffer containing 26% (v/v) ethanol placed in a reservoir. FeCl(3) and CO(3)(2-) in excess molar ratios to that of protein in its solution were added to the reservoir buffer. The crystals appeared after some hours and grew to the optimum size within 36 h. The structure was determined by molecular replacement method and refined to final R- and R-free factors of 0.187 and 0.255, respectively. The present structure showed that the protein molecule adopts an open conformation similar to that of camel apolactoferrin. The electron density map clearly indicated the presence of two iron atoms, one in each lobe with 4-fold coordinations: two by the protein ligands of Tyr-92(433) OH and Tyr-192(526) OH and two other coordination sites occupied by oxygen atoms of bidentate CO(3)(2-) ions leading to a tetrahedral intermediate. The CO(3)(2-) anion is stabilized through hydrogen bonds with the synergistic anion-binding site Arg-121(463) and with Ser-122 Ogamma in the N-lobe and Thr-464 Ogamma in C-lobe. The third oxygen atom of CO(3)(2-) interacts with a water molecule in both lobes. PMID- 11473114 TI - Human sex hormone-binding globulin promoter activity is influenced by a (TAAAA)n repeat element within an Alu sequence. AB - Sex hormone-binding globulin (SHBG) is the major sex steroid-binding protein in human plasma and is produced by the liver. Plasma SHBG levels vary considerably between individuals and are influenced by hormonal, metabolic, and nutritional factors. We have now found that a (TAAAA)(n) pentanucleotide repeat, located within an alu sequence at the 5' boundary of the human SHBG promoter, influences its transcriptional activity in association with downstream elements, including an SP1-binding site. Furthermore, SHBG alleles within the general population contain at least 6-10 TAAAA repeats, and the transcriptional activity of a human SHBG promoter-luciferase reporter construct containing 6 TAAAA repeats was significantly lower than for similar reporter constructs containing 7-10 TAAAA repeats when tested in human HepG2 hepatoblastoma cells. This difference in transcriptional activity reflected the preferential binding of a 46-kDa liver enriched nuclear factor to an oligonucleotide containing 6 rather than 7-10 TAAAA repeats. Thus, a (TAAAA)(n) element within the human SHBG promoter influences transcriptional activity in HepG2 cells and may contribute to differences in plasma SHBG levels between individuals. PMID- 11473115 TI - Identification of a critical lysine residue in apolipoprotein B-100 that mediates noncovalent interaction with apolipoprotein(a). AB - We have previously shown that lipoprotein(a) (Lp(a)) assembly involves an initial noncovalent interaction between sequences within apolipoprotein(a) (apo(a)) kringle IV types 5-8 and the amino terminus of apolipoprotein B-100 (sequences between amino acids 680 and 781 in apoB-100), followed by formation of a disulfide bond. In the present study, citraconylation of lysine residues in apoB 100 abolished the ability of the modified low density lipoprotein to associate with apo(a), thereby demonstrating a direct role for lysine residues in apoB in the first step of Lp(a) assembly. To identify specific lysine residues in the amino terminus of apoB that are required for the noncovalent interaction, we initially used an affinity chromatography method in which recombinant forms of apo(a) (r-apo(a)) were immobilized on Sepharose beads. Assessment of the ability of carboxyl-terminal truncations of apoB-18 to bind to r-apo(a)-Sepharose revealed that a 25-amino acid sequence in apoB (amino acids 680-704) bound specifically to apo(a) in a lysine-dependent manner; citraconylation of the lysine residues in the apoB derivative encoding this sequence abolished the binding interaction. Using fluorescence spectrometry, we found that a synthetic peptide corresponding to this sequence bound directly to apo(a); the peptide also reduced covalent Lp(a) formation. Lysine residues present in this sequence (Lys(680) and Lys(690)) were mutated to alanine in the context of apoB-18. We found that the apoB-18 species containing the Lys(680) mutation was incapable of binding to r-apo(a)-Sepharose columns, whereas the apoB-18 species containing the Lys(690) mutation exhibited slightly reduced binding to these columns. Taken together, our data indicate that Lys(680) is critical for the noncovalent interaction of apo(a) and apoB-100 that precedes covalent Lp(a) formation. PMID- 11473116 TI - Copper stabilizes a heterodimer of the yCCS metallochaperone and its target superoxide dismutase. AB - The copper chaperone for superoxide dismutase (CCS) activates the antioxidant enzyme Cu,Zn-SOD (SOD1) by directly inserting the copper cofactor into the apo form of SOD1. Neither the mechanism of protein-protein recognition nor of metal transfer is clear. The metal transfer step has been proposed to occur within a transient copper donor/acceptor complex that is either a heterodimer or heterotetramer (i.e. a dimer of dimers). To determine the nature of this intermediate, we generated a mutant form of SOD1 by replacing a copper binding residue His-48 with phenylalanine. This protein cannot accept copper from CCS but does form a stable complex with apo- and Cu-CCS, as observed by immunoprecipitation and native gel electrophoresis. Fluorescence anisotropy measurements corroborate the formation of this species and further indicate that copper enhances the stability of the dimer by an order of magnitude. The copper form of the heterodimer was isolated by gel filtration chromatography and contains one copper and one zinc atom per heterodimer. These results support a mechanism for copper transfer in which CCS and SOD1 dock via their highly conserved dimer interfaces in a manner that precisely orients the Cys-rich copper donor sites of CCS and the His-rich acceptor sites of SOD1 to form a copper bridged intermediate. PMID- 11473117 TI - Interaction of cationic antimicrobial peptides with model membranes. AB - A series of natural and synthetic cationic antimicrobial peptides from various structural classes, including alpha-helical, beta-sheet, extended, and cyclic, were examined for their ability to interact with model membranes, assessing penetration of phospholipid monolayers and induction of lipid flip-flop, membrane leakiness, and peptide translocation across the bilayer of large unilamellar liposomes, at a range of peptide/lipid ratios. All peptides were able to penetrate into monolayers made with negatively charged phospholipids, but only two interacted weakly with neutral lipids. Peptide-mediated lipid flip-flop generally occurred at peptide concentrations that were 3- to 5-fold lower than those causing leakage of calcein across the membrane, regardless of peptide structure. With the exception of two alpha-helical peptides V681(n) and V25(p,) the extent of peptide-induced calcein release from large unilamellar liposomes was generally low at peptide/lipid molar ratios below 1:50. Peptide translocation across bilayers was found to be higher for the beta-sheet peptide polyphemusin, intermediate for alpha-helical peptides, and low for extended peptides. Overall, whereas all studied cationic antimicrobial peptides interacted with membranes, they were quite heterogeneous in their impact on these membranes. PMID- 11473118 TI - GAD65-mediated glutamate decarboxylation reduces glucose-stimulated insulin secretion in pancreatic beta cells. AB - Mitochondrial metabolism plays a pivotal role in the pancreatic beta cell by generating signals that couple glucose sensing to insulin secretion. We have demonstrated previously that mitochondrially derived glutamate participates directly in the stimulation of insulin exocytosis. The aim of the present study was to impose altered cellular glutamate levels by overexpression of glutamate decarboxylase (GAD) to repress elevation of cytosolic glutamate. INS-1E cells infected with a recombinant adenovirus vector encoding GAD65 showed efficient overexpression of the GAD protein with a parallel increase in enzyme activity. In control cells glutamate levels were slightly increased by 7.5 mm glucose (1.4 fold) compared with the effect at 15 mm (2.3-fold) versus basal 2.5 mm glucose. Upon GAD overexpression, glutamate concentrations were no longer elevated by 15 mm glucose as compared with controls (-40%). Insulin secretion was stimulated in control cells by glucose at 7.5 mm (2.5-fold) and more efficiently at 15 mm (5.2 fold). INS-1E cells overexpressing GAD exhibited impaired insulin secretion on stimulation with 15 mm glucose (-37%). The secretory response to 30 mm KCl, used to raise cytosolic Ca(2+) levels, was unaffected. Similar results were obtained in perifused rat pancreatic islets following adenovirus transduction. This GAD65 mediated glutamate decarboxylation correlating with impaired glucose-induced insulin secretion is compatible with a role for glutamate as a glucose-derived factor participating in insulin exocytosis. PMID- 11473119 TI - Recruitment of multiple interferon regulatory factors and histone acetyltransferase to the transcriptionally active interferon a promoters. AB - Type I interferon (IFN) plays a critical role in the innate immunity against viral infection. Expression of IFNA genes in infected cells is cell type dependent and is regulated at the transcriptional level. The present study is focused on the molecular mechanism underlying the differential expression of human IFNA1 and A2 genes. Two nucleotides, at positions -98 and -81 of IFNA1 and A2 promoter, were pivotal to the differential expression. The DNA pull-down and chromatin precipitation assays have shown that nuclear interferon regulatory factor (IRF)-3 and IRF-7 as well as IRF-1 bind to IFNA1 virus-responsive element (VRE). Interestingly, overexpression of IRF-7 increased the otherwise weak binding of both IRF-3 and IRF-7 to IFNA2 VRE. These data together with the results of two-step chromatin immunoprecipitation strongly suggest that the IRF-3 and IRF-7 bind to IFNA1 promoter as a dimer. Furthermore, binding of IRF-3 and IRF-7 to IFNA VRE is associated with the presence of acetylated histone H3, suggesting that histone acetyltransferase(s) is tethered together with virus activated IRF-3 and IRF-7 to the IFNA1 promoter. In addition, the constitutively active IRF-3 (5D) and IRF-7 (2D) mutants activate the endogenous IFNA genes in uninfected cells; however, the expression profile of IFNA is not identical to that induced by viral infection. PMID- 11473120 TI - Mapping the interacting regions between troponins T and C. Binding of TnT and TnI peptides to TnC and NMR mapping of the TnT-binding site on TnC. AB - Muscular contraction is triggered by an increase in calcium concentration, which is transmitted to the contractile proteins by the troponin complex. The interactions among the components of the troponin complex (troponins T, C, and I) are essential to understanding the regulation of muscle contraction. While the structure of TnC is well known, and a model for the binary TnC.TnI complex has been recently published (Tung, C.-S., Wall, M. E., Gallagher, S. C., and Trewhella, J. (2000) Protein Sci. 9, 1312-1326), very little is known about TnT. Using non-denaturing gels and NMR spectroscopy, we have analyzed the interactions between TnC and five peptides from TnT as well as how three TnI peptides affect these interactions. Rabbit fast skeletal muscle peptide TnT-(160-193) binds to TnC with a dissociation constant of 30 +/- 6 microm. This binding still occurs in the presence of TnI-(1-40) but is prevented by the presence of TnI-(56-115) or TnI-(96-139), both containing the primary inhibitory region of TnI. TnT-(228-260) also binds TnC. The binding site for TnT-(160-193) is located on the C-terminal domain of TnC and was mapped to the surface of TnC using NMR chemical shift mapping techniques. In the context of the model for the TnC.TnI complex, we discuss the interactions between TnT and the other troponin subunits. PMID- 11473121 TI - Cleavage of doublecortin-like kinase by calpain releases an active kinase fragment from a microtubule anchorage domain. AB - Doublecortin-like kinase (DCLK) is widely expressed in postmitotic neurons throughout the embryonic nervous system. DCLK consists of an N-terminal doublecortin domain, responsible for its localization to microtubules, and a C terminal serine-threonine kinase domain. Here we report that DCLK is a physiological substrate for the cysteine protease calpain. Cleavage of DCLK by calpain severs the kinase domain from its microtubule anchorage domain and releases it into the cytoplasm. The isolated kinase domain retains catalytic activity and is structurally similar to CPG16, a second product of the DCLK gene expressed in the adult brain that lacks the doublecortin domain. We propose that in neurons cleavage of DCLK by calpain represents a calcium responsive mechanism to regulate localization of the DCLK kinase domain. PMID- 11473122 TI - Fibroblast growth factor-2 antagonist activity and angiostatic capacity of sulfated Escherichia coli K5 polysaccharide derivatives. AB - The angiogenic basic fibroblast growth factor (FGF2) interacts with tyrosine kinase receptors (FGFRs) and heparan sulfate proteoglycans (HSPGs) in endothelial cells. Here, we report the FGF2 antagonist and antiangiogenic activity of novel sulfated derivatives of the Escherichia coli K5 polysaccharide. K5 polysaccharide was chemically sulfated in N- and/or O-position after N-deacetylation. O-Sulfated and N,O-sulfated K5 derivatives with a low degree and a high degree of sulfation compete with heparin for binding to 125I-FGF2 with different potency. Accordingly, they abrogate the formation of the HSPG.FGF2.FGFR ternary complex, as evidenced by their capacity to prevent FGF2-mediated cell-cell attachment of FGFR1-overexpressing HSPG-deficient Chinese hamster ovary (CHO) cells to wild type CHO cells. They also inhibited 125I-FGF2 binding to FGFR1-overexpressing HSPG-bearing CHO cells and adult bovine aortic endothelial cells. K5 derivatives also inhibited FGF2-mediated cell proliferation in endothelial GM 7373 cells and in human umbilical vein endothelial (HUVE) cells. In all these assays, the N sulfated K5 derivative and unmodified K5 were poorly effective. Also, highly O sulfated and N,O-sulfated K5 derivatives prevented the sprouting of FGF2 transfected endothelial FGF2-T-MAE cells in fibrin gel and spontaneous angiogenesis in vitro on Matrigel of FGF2-T-MAE and HUVE cells. Finally, the highly N,O-sulfated K5 derivative exerted a potent antiangiogenic activity on the chick embryo chorioallantoic membrane. These data demonstrate the possibility of generating FGF2 antagonists endowed with antiangiogenic activity by specific chemical sulfation of bacterial K5 polysaccharide. In particular, the highly N,O sulfated K5 derivative may provide the basis for the design of novel angiostatic compounds. PMID- 11473123 TI - Crystal structure of the FAD/NADPH-binding domain of rat neuronal nitric-oxide synthase. Comparisons with NADPH-cytochrome P450 oxidoreductase. AB - Nitric-oxide synthase (NOS) is composed of a C-terminal, flavin-containing reductase domain and an N-terminal, heme-containing oxidase domain. The reductase domain, similar to NADPH-cytochrome P450 reductase, can be further divided into two different flavin-containing domains: (a) the N terminus, FMN-containing portion, and (b) the C terminus FAD- and NADPH-binding portion. The crystal structure of the FAD/NADPH-containing domain of rat neuronal nitric-oxide synthase, complexed with NADP(+), has been determined at 1.9 A resolution. The protein is fully capable of reducing ferricyanide, using NADPH as the electron donor. The overall polypeptide fold of the domain is very similar to that of the corresponding module of NADPH-cytochrome P450 oxidoreductase (CYPOR) and consists of three structural subdomains (from N to C termini): (a) the connecting domain, (b) the FAD-binding domain, and (c) the NADPH-binding domain. A comparison of the structure of the neuronal NOS FAD/NADPH domain and CYPOR reveals the strict conservation of the flavin-binding site, including the tightly bound water molecules, the mode of NADP(+) binding, and the aromatic residue that lies at the re-face of the flavin ring, strongly suggesting that the hydride transfer mechanisms in the two enzymes are very similar. In contrast, the putative FMN domain-binding surface of the NOS protein is less positively charged than that of its CYPOR counterpart, indicating a different nature of interactions between the two flavin domains and a different mode of regulation in electron transfer between the two flavins involving the autoinhibitory element and the C-terminal 33 residues, both of which are absent in CYPOR. PMID- 11473124 TI - A SecY homologue is involved in chloroplast-encoded D1 protein biogenesis. AB - We have used the photosystem II reaction center D1 protein as a model to study the mechanisms of targeting and insertion of chloroplast-encoded thylakoid membrane proteins. The unusually high turnover rate and distinct pausing intermediates during translation make the D1 protein biogenesis particularly suitable for these purposes. Here we show that cpSecY, a chloroplast homologue of bacterial essential translocon component SecY, interacts tightly with thylakoid membrane-bound ribosomes, suggesting its involvement in protein translocation and insertion. Co-immunoprecipitation and cross-linking experiments indicated that cpSecY resides in the vicinity of D1 elongation intermediates and provided evidence for a transient interaction of cpSecY with D1 elongation intermediates during the biogenesis of D1. After termination of translation, such interactions no longer existed. Our results indicate that, in addition to a well characterized role of cpSecY in posttranslational translocation of nuclear-encoded proteins, it seems to be also involved in cotranslational membrane protein translocation and insertion in chloroplasts. PMID- 11473125 TI - HMG-D and histone H1 interplay during chromatin assembly and early embryogenesis. AB - HMG-D is an abundant chromosomal protein associated with condensed chromatin during the first nuclear cleavage cycles of the developing Drosophila embryo. We previously suggested that HMG-D might substitute for the linker histone H1 in the preblastoderm embryo and that this substitution might result in the characteristic less compacted chromatin. We have now studied the association of HMG-D with chromatin using a cell-free system for chromatin reconstitution derived from Drosophila embryos. Association of HMG-D with chromatin, like that of histone H1, increases the nucleosome spacing indicative of binding to the linker DNA between nucleosomes. HMG-D interacts with DNA during the early phases of nucleosome assembly but is gradually displaced as chromatin matures. By contrast, purified chromatin can be loaded with stoichiometric amounts of HMG-D, and this can be displaced upon addition of histone H1. A direct physical interaction between HMG-D and histone H1 was observed in a Far Western analysis. The competitive nature of this interaction is reminiscent of the apparent replacement of HMG-D by H1 during mid-blastula transition. These data are consistent with the hypothesis that HMG-D functions as a specialized linker protein prior to appearance of histone H1. PMID- 11473126 TI - Roles for beta(pat-3) integrins in development and function of Caenorhabditis elegans muscles and gonads. AB - Heterodimeric integrin receptors for extracellular matrix (ECM) play vital roles in bidirectional signaling during tissue development, organization, remodeling, and repair. The beta integrin subunit cytoplasmic domain is essential for transmission of many of these signals and overexpression of an unpaired beta tail in cultured cells inhibits endogenous integrins. Unlike vertebrates, which have at least nine beta subunit genes, the nematode Caenorhabditis elegans expresses only one beta subunit (betapat-3), and a null mutation in this gene causes embryonic lethality. To determine the functions of integrins during larval development and in adult tissues, we have taken a dominant negative approach by expression of an HA-betatail transgene composed of a hemagglutinin (HA) epitope tag extracellular domain connected to the betapat-3 transmembrane and cytoplasmic domains. Expression of this transgene in muscle and gonad, major sites of integrin expression, caused a variety of phenotypes dependent on the level of transgene expression. Abnormalities in body wall and sex muscles led to uncoordinated movement and egg-laying defects. Significant anomalies in migration and pathfinding were caused by tissue-specific expression of HA-betatail in the distal tip cells (DTC), the cells that direct gonad morphogenesis. A pat-3 gene with Tyr to Phe mutations in the cytoplasmic domain was able to rescue pat-3 null animals but also showed DTC migration defects. These results show that betapat-3 plays important roles in post-embryonic organogenesis and tissue function. PMID- 11473127 TI - Evidence that ligand and metal ion binding to integrin alpha 4beta 1 are regulated through a coupled equilibrium. AB - We have used the highly selective alpha(4)beta(1) inhibitor 2S-[(1 benzenesulfonyl-pyrrolidine-2S-carbonyl)-amino]-4-[4-methyl-2S-(methyl-[2-[4-(3-o tolyl-ureido)-phenyl]-acetyl]-amino)-pentanoylamino]-butyric acid (BIO7662) as a model ligand to study alpha(4)beta(1) integrin-ligand interactions on Jurkat cells. Binding of [(35)S]BIO7662 to Jurkat cells was dependent on the presence of divalent cations and could be blocked by treatment with an excess of unlabeled inhibitor or with EDTA. K(D) values for the binding of BIO7662 to Mn(2+) activated alpha(4)beta(1) and to the nonactivated state of the integrin that exists in 1 mm Mg(2+), 1 mm Ca(2+) were <10 pm, indicating that it has a high affinity for both activated and nonactivated integrin. No binding was observed on alpha(4)beta(1) negative cells. Through an analysis of the metal ion dependences of ligand binding, several unexpected findings about alpha(4)beta(1) function were made. First, we observed that Ca(2+) binding to alpha(4)beta(1) was stimulated by the addition of BIO7662. From solution binding studies on purified alpha(4)beta(1), two types of Ca(2+)-binding sites were identified, one dependent upon and the other independent of BIO7662 binding. Second, we observed that the metal ion dependence of ligand binding was affected by the affinity of the ligand for alpha(4)beta(1). ED(50) values for the metal ion dependence of the binding of BIO7762 and the binding of a lower affinity ligand, BIO1211, differed by 2-fold for Mn(2+), 30-fold for Mg(2+), and >1000-fold for Ca(2+). Low Ca(2+) (ED(50) = 5 10 microm) stimulated the binding of BIO7662 to alpha(4)beta(1). The effects of microm Ca(2+) closely resembled the effects of Mn(2+) on alpha(4)beta(1) function. Third, we observed that the rate of BIO7662 binding was dependent on the metal ion concentration and that the ED(50) for the metal ion dependence of BIO7662 binding was affected by the concentration of the BIO7662. These studies point to an even more complex interplay between metal ion and ligand binding than previously appreciated and provide evidence for a three-component coupled equilibrium model for metal ion-dependent binding of ligands to alpha(4)beta(1). PMID- 11473128 TI - Biochemical characterization and ligand binding properties of neuroglobin, a novel member of the globin family. AB - Neuroglobin is a recently discovered member of the globin superfamily that is suggested to enhance the O(2) supply of the vertebrate brain. Spectral measurements with human and mouse recombinant neuroglobin provide evidence for a hexacoordinated deoxy ferrous (Fe(2+)) form, indicating a His-Fe(2+)-His binding scheme. O(2) or CO can displace the endogenous protein ligand, which is identified as the distal histidine by mutagenesis. The ferric (Fe(3+)) form of neuroglobin is also hexacoordinated with the protein ligand E7-His and does not exhibit pH dependence. Flash photolysis studies show a high recombination rate (k(on)) and a slow dissociation rate (k(off)) for both O(2) and CO, indicating a high intrinsic affinity for these ligands. However, because the rate-limiting step in ligand combination with the deoxy hexacoordinated form involves the dissociation of the protein ligand, O(2) and CO binding is suggested to be slow in vivo. Because of this competition, the observed O(2) affinity of recombinant human neuroglobin is average (1 torr at 37 degrees C). Neuroglobin has a high autoxidation rate, resulting in an oxidation at 37 degrees C by air within a few minutes. The oxidation/reduction potential of mouse neuroglobin (E'(o) = -129 mV) lies within the physiological range. Under natural conditions, recombinant mouse neuroglobin occurs as a monomer with disulfide-dependent formation of dimers. The biochemical and kinetic characteristics are discussed in view of the possible functions of neuroglobin in the vertebrate brain. PMID- 11473129 TI - Metabolism of sucrose and its five linkage-isomeric alpha-D-glucosyl-D-fructoses by Klebsiella pneumoniae. Participation and properties of sucrose-6-phosphate hydrolase and phospho-alpha-glucosidase. AB - Klebsiella pneumoniae is presently unique among bacterial species in its ability to metabolize not only sucrose but also its five linkage-isomeric alpha-d glucosyl-d-fructoses: trehalulose, turanose, maltulose, leucrose, and palatinose. Growth on the isomeric compounds induced a protein of molecular mass approximately 50 kDa that was not present in sucrose-grown cells and which we have identified as an NAD(+) and metal ion-dependent 6-phospho-alpha-glucosidase (AglB). The aglB gene has been cloned and sequenced, and AglB (M(r) = 49,256) has been purified from a high expression system using the chromogenic p-nitrophenyl alpha-glucopyranoside 6-phosphate as substrate. Phospho-alpha-glucosidase catalyzed the hydrolysis of a wide variety of 6-phospho-alpha-glucosides including maltose-6'-phosphate, maltitol-6-phosphate, isomaltose-6'-phosphate, and all five 6'-phosphorylated isomers of sucrose (K(m) approximately 1-5 mm) yet did not hydrolyze sucrose-6-phosphate. By contrast, purified sucrose-6-phosphate hydrolase (M(r) approximately 53,000) hydrolyzed only sucrose-6-phosphate (K(m) approximately 80 microm). Differences in molecular shape and lipophilicity potential between sucrose and its isomers may be important determinants for substrate discrimination by the two phosphoglucosyl hydrolases. Phospho-alpha glucosidase and sucrose-6-phosphate hydrolase exhibit no significant homology, and by sequence-based alignment, the two enzymes are assigned to Families 4 and 32, respectively, of the glycosyl hydrolase superfamily. The phospho-alpha glucosidase gene (aglB) lies adjacent to a second gene (aglA), which encodes an EII(CB) component of the phosphoenolpyruvate-dependent sugar:phosphotransferase system. We suggest that the products of the two genes facilitate the phosphorylative translocation and subsequent hydrolysis of the five alpha-d glucosyl-d-fructoses by K. pneumoniae. PMID- 11473130 TI - Differential regulation of the dopamine D2 and D3 receptors by G protein-coupled receptor kinases and beta-arrestins. AB - The D(2) and D(3) receptors (D(2)R and D(3)R), which are potential targets for antipsychotic drugs, have a similar structural architecture and signaling pathway. Furthermore, in some brain regions they are expressed in the same cells, suggesting that differences between the two receptors might lie in other properties such as their regulation. In this study we investigated, using COS-7 and HEK-293 cells, the mechanism underlying the intracellular trafficking of the D(2)R and D(3)R. Activation of D(2)R caused G protein-coupled receptor kinase dependent receptor phosphorylation, a robust translocation of beta-arrestin to the cell membrane, and profound receptor internalization. The internalization of the D(2)R was dynamin-dependent, suggesting that a clathrin-coated endocytic pathway is involved. In addition, the D(2)R, upon agonist-mediated internalization, localized to intracellular compartments distinct from those utilized by the beta(2)-adrenergic receptor. However, in the case of the D(3)R, only subtle agonist-mediated receptor phosphorylation, beta-arrestin translocation to the plasma membrane, and receptor internalization were observed. Interchange of the second and third intracellular loops of the D(2)R and D(3)R reversed their phenotypes, implicating these regions in the regulatory properties of the two receptors. Our studies thus indicate that functional distinctions between the D(2)R and D(3)R may be found in their desensitization and cellular trafficking properties. The differences in their regulatory properties suggest that they have distinct physiological roles in the brain. PMID- 11473131 TI - Potent regulation of midbrain dopamine neurons by the bed nucleus of the stria terminalis. AB - Recent studies have revealed an important role of the ventrolateral (subcommissural) aspect of the bed nucleus of the stria terminalis (vBNST) in motivational aspects of drug abuse (Delfs et al., 2000). Dopaminergic (DA) neurons in the ventral tegmental area (VTA) have also long been linked to motivation and drug abuse (Koob and Le Moal, 2001). The present study tested whether activity in the vBNST influences discharge of midbrain DA neurons. Responses of DA neurons in the VTA to activation of the vBNST were characterized in anesthetized rats using extracellular recording techniques. Electrical or chemical [10-50 mm glutamate (Glu)] stimulation of the vBNST consistently activated DA cells (122% increase in activity with 50 mm Glu). However, stronger chemical stimulation of the vBNST (100 mm Glu) completely inactivated DA neurons. In addition, apomorphine restored the activity of DA neurons that were inactivated by 100 mm Glu stimulation of the vBNST, indicating possible depolarization blockade of DA cells by vBNST activity. These findings reveal that the vBNST exerts a strong excitatory influence on DA neurons. Also striking was the finding that chemical stimulation (50 mm Glu) of the vBNST yielded long lasting oscillatory activity (>15 min) in VTA DA neurons. These results indicate that the vBNST can generate long-lasting alterations in the activity of DA neurons in vivo. PMID- 11473132 TI - Synapse formation between central neurons requires postsynaptic expression of the MEN1 tumor suppressor gene. AB - Synapse formation is a crucial step in the development of neuronal circuits and requires precise coordination of presynaptic and postsynaptic activities. However, molecular mechanisms that control the formation of functionally mature synaptic contacts, in particular between central neurons, remain poorly understood. To identify genes that are involved in the formation of central synapses, we made use of molluscan neurons that in culture form synaptic contacts between their somata (soma-soma synapses) in the absence of neurite outgrowth. Using single-cell mRNA differential display, we have identified a molluscan homolog of the multiple endocrine neoplasia type 1 (MEN1) tumor suppressor gene encoding the transcription factor menin as a gene that is upregulated during synapse formation. In vitro antisense knock-down of MEN1 mRNA blocks the formation of mature synapses between different types of identified central neurons. Moreover, immunocytochemistry and cell-specific knock-down of MEN1 mRNA show that postsynaptic but not presynaptic expression is required for synapses to form. Together, our data demonstrate that menin is a synaptogenic factor that is critically involved in a general postsynaptic mechanism of synapse formation between central neurons. PMID- 11473133 TI - Mitogen-activated protein kinase cascade in the basolateral nucleus of amygdala is involved in extinction of fear-potentiated startle. AB - Previous results indicate that intra-amygdala infusions of NMDA receptor antagonists block the extinction of conditioned fear. Mitogen-activated protein kinase (MAPK) can be activated by NMDA receptor stimulation and is involved in excitatory fear conditioning. Here, we evaluate the role of MAPK within the basolateral amygdala in the extinction of conditioned fear. Rats received 10 light-shock pairings. After 24 hr, fear was assessed by eliciting the acoustic startle reflex in the presence of the conditioned stimulus (CS) (CS-noise trials) and also in its absence (noise-alone trials). Rats subsequently received an intra amygdala or intrahippocampal infusion of either 20% DMSO or the MAPK inhibitor PD98059 (500 ng/side) followed 10 min later by 30 presentations of the light CS without shock (extinction training). After 24 hr, they were again tested for fear potentiated startle. PD98059 infusions into the basolateral amygdala but not the hippocampus significantly reduced extinction, which was otherwise evident in DMSO infused rats. Control experiments indicated that the effect of intra-amygdala PD98059 could not be attributed to lasting damage to the amygdala or to state dependency. These results suggest that a MAPK-dependent signaling cascade within or very near the basolateral amygdala plays an important role in the extinction of conditioned fear. PMID- 11473134 TI - Learning from teachers. PMID- 11473135 TI - Occupational injuries to fisheries workers in Norway reported to insurance companies from 1991 to 1996. AB - Fisheries work is one of the occupations at highest risk for occupational accidents in many countries. It is necessary to understand the injuries in order to prevent them. This study of occupational injury claims by fisheries workers in Norway made to insurance companies from 1991 to 1996 analysed the workers' age, time of injury, injury type, part of the body involved, injury event and cost. The highest injury incidence rates were among the younger fisheries workers and during the winter months. Bruises and fractures were the most frequent injury types, and fingers and hands were most often affected, whereas falls and accidents related to machines were the most common causes. Safety measures should be taken on board to prevent falls and machine-related injuries, and young fisheries workers should have better on-the-job training. PMID- 11473136 TI - The health of fishermen in the catching sector of the fishing industry: a gap analysis. AB - Offshore fishing is a sizeable industry in the UK and it is one of the most dangerous occupations. Long hours, extreme weather and working with heavy machinery contribute to a high mortality rate in fishermen. Despite a long standing acknowledgement of the dangers and high mortality associated with fishing, there has been little research in this field. Although there have been developments within the industry in terms of safety, there has been little emphasis on the relationship between health, the environment and performance at work. This paper reviews the international literature on the health of fishermen, and describes the main findings and highlights gaps in current evidence. Areas for further research that would inform the future development of an evidence based occupational health service for the offshore fishing industry are discussed. PMID- 11473137 TI - Occupational health in general practice in an industrial area of Singapore. AB - The aim of this study was to estimate the caseload with regard to the proportion of work-related medical cases in factory workers and to survey knowledge of and attitudes to occupational medicine among doctors in an industrial area in Singapore. To this end, a self-administered questionnaire was given to 74 general practitioners in the industrial district of Jurong in Singapore. Overall, there was an 89.2% response rate. Thirty-three of the 66 respondents were males. Mean age was 40.8 years (range 27-64 years). Twenty-four (36%) had some postgraduate training in occupational medicine (GP-OM) while the rest did not (GP). The caseload for all doctors was similar with regard to the number of adult patients and, more specifically, the number of factory workers seen per day. The majority of doctors (76.6%) felt that <10% of factory workers seen had work-related problems. Most (70%) felt confident in dealing with the occupational problems that arose. However, a large percentage (78% of GPs, 45.8% of GP-OMs) felt that training in occupational medicine was inadequate. This is reflected in the knowledge questions, where GPs did not score as well as GP-OMs. It was concluded that GPs working in this industrial area see a good proportion of factory workers, in a fair number of cases for work-related problems. However, only a third of the GPs are qualified in occupational medicine. These factors highlight the need for more emphasis on occupational medicine training among general practitioners, especially those working in industrial areas. PMID- 11473138 TI - Promotion of work ability, the quality of work and retirement. AB - In this study, the validity of a model designed to promote the work ability of aging workers was examined. The target areas of work ability promotion were searched for the characteristics that explain work ability the best. In addition, the way work ability relates to the quality of work and retirement was examined. The subjects (n = 1101) participated in the follow-up study on aging Finnish workers in 1992 and 1997. The results consistently supported the model for promoting work ability. All four areas of focus-(i) work demands and the environment; (ii) work organization and the work community; (iii) the promotion of workers' health and functional capacity; and (iv) the promotion of professional competence-proved to be strongly associated with work ability. Good work ability was associated with a high quality of work and the enjoyment of staying in one's job. It also predicted active and meaningful retirement. PMID- 11473139 TI - Estimating occupational health events in workers with asthma or diabetes: a comparison of diary and snapshot methods. AB - The study was set up to assess the completeness of reporting different types of work-related health events through 6 month and 1 week diaries and telephone questionnaires relating to individual days. Subjects recruited from hospital asthma or diabetic clinics or from surgical day-case units were randomly assigned to either a full participation arm or to one completing only a 6 month diary. Of the 375 study participants, return rates were 68.8% for 6 month diaries, 81.9% for weekly diaries and 86.5% for researcher-administered questionnaires. Significantly higher rates of self-reported poor work performance, symptoms or changes in medication were estimated by snapshot methods (same-day telephone interviews or one-off weekly diaries) than from diaries completed over a 6 month period. Asthmatics and diabetics attending hospital outpatient clinics were no more likely, however, to report work-related health events in the following 6 months than those attending the same hospital for day surgery. PMID- 11473140 TI - Pre-employment health assessments for food handlers: a survey of occupational physicians in the food industry. AB - The format of pre-employment health screens within the food industry varies considerably. The aim of this study was to produce a consensus on the content of a screen that will enable employees to handle food with minimum risk to the product and employee. A questionnaire was sent to the 63 members of the Food Industry Medical Association, of whom 45 (71%) responded. As a minimum, a questionnaire completed by the applicant is thought to be sufficient. Applicants reporting health problems need to be assessed by a health professional. PMID- 11473141 TI - Upper aerodigestive tract cancers in former employees at an iron and steel works. AB - Local concern about numbers of laryngeal cancer cases led to an investigation of the incidence of upper aerodigestive tract (UAT) cancer in an industrial cohort. Males (n = 11 470) who had been directly employed at an iron and steel works in northern England at any time between January 1960 and site closure in September 1983 were followed up for UAT cancers until December 1998. The incidence of UAT and laryngeal cancer was compared to the general population of the region via indirect standardization. Fifty-two members of the cohort developed a UAT cancer during 1960-1998. There were no more UAT cancers than expected [standardized incidence ratio = 97, 95% confidence interval (CI) = 72-127], but slightly more laryngeal cancers than expected (standardized incidence ratio = 118, 95% CI = 78 171), although this estimate was less precise. The lack of complete work histories meant that relationships between cancer incidence and length of service or job categories could not be explored. PMID- 11473142 TI - Colorectal cancer among Chinese restaurant waiters. AB - To investigate whether Chinese restaurant waiters in Hong Kong had an increased risk of dying from colorectal cancer, we linked the records of deceased union members to the mortality information for the general population between 1986 and 1995. A case-control analysis among the deceased was performed with 346 deceased waiters and 306 deceased male kitchen workers. Waiters were found to have a significantly higher mortality odds ratio [(MOR) 2.29; 95% confidence interval (CI) = 1.47-3.56] of colorectal cancer when compared with the general population. The MOR (1.45; 95% CI = 0.71-2.93) was increased, but not significantly, when compared with the kitchen workers. The study suggested that waiters in Hong Kong Chinese restaurants are at a higher risk of dying from colorectal cancer. An unhealthy diet associated with their job is a possible explanation, and interventions should be initiated. PMID- 11473143 TI - Long-term sickness absence - risk indicators among occupationally active residents of a Danish county. AB - This study aims to identify risk factors and their prevalence in long-term sickness absence. The study is designed as a case-referent study which comprises 481 participants who have experienced a sickness absence lasting >10 weeks and a reference group of 1326 individuals in active employment. Multivariate analysis identified the following significant risk factors for men: (i) age >50 years [odds ratio (OR) = 2.4]; (ii) short period of education (OR = 2.3); (iii) unemployment within the last 3 years (OR = 1.7); (iv) heavy-duty work (OR = 2.1); (v) monotonous, repetitive work (OR = 1.7); (vi) lack of job satisfaction (OR = 2.1); and (vii) much back pain during the last 3 years (OR = 2.1). The following risk factors were identified for women: (i) leaving school without graduation (OR = 2.6); (ii) unemployment within the last 3 years (OR = 1.5); (iii) heavy-duty work (OR = 2.8); (iv) lack of influence on own job situation (OR = 2.1); and (v) much back pain within the last 3 years (OR = 1.8). It is concluded that the identification of working environment risk factors constitutes a case for improvement of the working environment which may be instrumental in reducing long term sickness absence. PMID- 11473144 TI - A risk-based system to penalize and reward line management for occupational safety and health performance. AB - Penalizing line management for the occurrence of lost time injuries has in some cases had unintended negative consequences. These are discussed. An alternative system is suggested that penalizes line management for accidents where the combination of the probability of recurrence and the maximum reasonable consequences such a recurrence may have exceeds an agreed limit. A reward is given for prompt effective control of the risk to below the agreed risk limit. The reward is smaller than the penalty. High-risk accidents require independent investigation by a safety officer using analytical techniques. Two case examples are given to illustrate the system. Continuous safety improvement is driven by a planned reduction in the agreed risk limit over time and reward for proactive risk assessment and control. PMID- 11473145 TI - Assessment of physical fitness for occupations encompassing load-carriage tasks. AB - There is increasing anecdotal evidence that simple occupational tests of aerobic fitness impose a systematic bias against heavier personnel when predicting fitness for load-carrying tasks. This study tested the hypothesis that simple field tests of aerobic fitness are not good predictors of load-carrying performance and that personnel with greater body mass are more able to perform occupationally relevant load-carrying tasks. Twelve healthy male volunteers ran on a level treadmill at 9.5 km/h for 4 min, with (T18) and without (T0) an external backpack load of 18 kg. During each exercise period, steady-state oxygen uptake (VO(2)) was assessed. On a subsequent occasion (at least 7 days later), 11 of the subjects ran to exhaustion at 9.5 km/h whilst carrying the 18 kg external load (ETT18). There was a strong inverse linear relationship between relative VO(2) and body mass (r = -0.87, P < 0.01) and between VO(2) and lean body mass (r = -0.74, P < 0.01) during the T18 trials. Furthermore, there was a moderately strong relationship between exercise time (ETT18) and body mass (r = 0.69, P < 0.05) and between exercise time and lean body mass (r = 0.71, P < 0.05). There was no relationship between exercise tolerance time and VO(2) (r = 0.12). The results show that fitness tests that determine aerobic power in units relative to body mass (e.g. timed distance run) incur a systematic bias against heavier personnel. Such tests are therefore inappropriate when predicting the ability of personnel to work in occupations that encompass load-carrying tasks. PMID- 11473146 TI - Perforation of the nasal septum. PMID- 11473153 TI - Cellular changes in transplanted human corneas. AB - PURPOSE: To measure endothelial cell and keratocyte densities in transplanted corneas and the changes in these densities with time. METHODS: The endothelia of 500 consecutive penetrating corneal transplants were studied longitudinally by specular microscopy for 10 to 20 years. The keratocytes of 36 corneal transplants that varied in postoperative times from 1 month to 20 years were studied cross sectionally by clinical confocal microscopy. The keratocytes of five transplanted corneas were studied longitudinally by confocal microscopy at 1 day, 1 week, and 1 month postkeratoplasty. RESULTS: Endothelial cell density decreased progressively at an accelerated rate for 20 years after transplantation, with concurrent increases in the coefficient of variation of cell area and corneal thickness and decreases in the percentage of hexagonal cells. Grafts with insufficient endothelial cells developed late endothelial failure, which was the primary cause of graft failure after the first 5 postoperative years. The grafts with late endothelial failure did not lose endothelial cells faster than grafts that did not fail, but instead had fewer cells immediately after transplantation, diminishing to a critically low cell density earlier. The keratocyte density was also decreased in transplanted corneas. Keratocytes became "activated" during the first week after keratoplasty and in grafts with late endothelial failure. CONCLUSION: It should be possible to prevent or delay late endothelial failure, the primary cause of graft failure, by increasing the number of endothelial cells on transplanted corneas. The status of the keratocytes appears to affect corneal transparency and, thus, visual quality in the grafted eye. PMID- 11473154 TI - Anterior stromal puncture in the treatment of bullous keratopathy: six-month follow-up. AB - PURPOSE: This study was designed to evaluate the effect of anterior stromal puncture to treat patients with symptomatic bullous keratopathy. METHODS: Fourteen patients with bullous keratopathy who had decreased vision and were or were not awaiting penetrating keratoplasty were evaluated before and 1, 4, 12, and 24 weeks after anterior stromal punctures with a 25-gauge needle. The patients were questioned about pain intensity, foreign-body sensation, and photophobia. Biomicroscopy, esthesiometry, and pachymetry results were evaluated. The results before and after the punctures were compared using the McNemar test, Friedman test, and multiple comparison tests. RESULTS: The comparisons among before and 1, 4, and 12 weeks after the anterior stromal puncture of pain (p = 0.00000556), photophobia (p = 0.0252), foreign-body sensation (p = 0.0000743), and esthesiometry (p = 0.0243) showed significant statistical differences in terms of decreasing symptoms and corneal sensitivity. The average pachymetry did not show statistical difference among before and 1, 4, 12, and 24 weeks after the procedure (p = 0.956). We did not find important differences in terms of corneal neovascularization. CONCLUSION: Anterior stromal puncture is an effective, simple, and low-cost alternative for treating patients with symptomatic bullous keratopathy. PMID- 11473155 TI - Anterior stromal puncture in bullous keratopathy: a clinicopathologic study. AB - PURPOSE: We report our experience of anterior stromal puncture (ASP) in symptomatic chronic corneal edema patients awaiting penetrating keratoplasty (PK) and hypothesize the mechanism of action of ASP in bullous keratopathy (BK). METHODS: We reviewed the medical records of 28 patients with BK who underwent ASP between November 1996 and September 1999 with at least 1 month of follow-up. In all these patients, approximately 200 punctures were given with a bent 26-gauge needle sparing the peripheral cornea. Patients were examined on days 1 and 7, 1 month, and every 3 months after the procedure. The symptoms, corneal findings, and visual acuity of the patients at last follow-up were noted and compared with the preprocedure findings. For 11 patients who underwent PK, histopathologic study of corneal buttons was performed and a clinicopathologic correlation was attempted. RESULTS: Of the 28 patients, there were 15 men and 13 women with a mean age of 61.1 years. The clinical diagnosis was pseudophakic BK in 11 (39.3%), aphakic BK in 11 (39.3%), Fuchs' dystrophy in 4 (14.3%), failed graft in 1 (3.6%), and chronic corneal edema of unknown etiology in 1 (3.6%). The follow-up ranged from 1 to 33 months with a mean of 9.5 +/- 7.5 months. Symptomatic relief was noted in all. Twenty patients (71.4%) had complete relief, whereas eight patients (28.6%) experienced mild symptoms such as tearing and occasional pain. Visual acuity improved in 7 patients (25%), decreased in 12 (42.8%), and remained the same in 9 (32.4%). Objective evidence of scarring after ASP was noted in all patients. Complete regression of epithelial bullae and epithelial edema was found in 10 (35.7%) and partial regression in 18 (62.25%) patients. There was no progression or appearance of new blood vessels except in one patient (3.6%). Histologically, puncture marks and superficial stromal scarring were noted in all corneal buttons. Adhesion of epithelium with varying degrees of subepithelial fibrosis was seen in six, whereas in the remaining five buttons, there was complete denudation of the epithelium. Vascularization was seen in five and inflammation in two buttons. Clinicopathologic correlation revealed that symptomatic patients had persistent edema and loose adhesion of the epithelium to the stroma, resulting in detachment. CONCLUSIONS: ASP is a simple, safe, and cost effective outpatient procedure for symptomatic relief in patients with BK. The possibility of decreased visual acuity after the procedure should be explained to all patients. Although ASP promoted subepithelial fibrosis in all cases, its subsequent adhesion is variable and probably has clinical relevance. Further studies could be directed toward identifying specific mediators that promote epithelial-stromal interaction and firm anchoring of epithelium to the underlying stroma. PMID- 11473156 TI - Intraocular lens implantation after penetrating keratoplasty. AB - PURPOSE: We evaluated the clinical results of nonsimultaneous penetrating keratoplasty, cataract extraction, and intraocular lens (IOL) implantation (two stage surgery) for combined corneal disease and cataract. METHODS: Twenty-six eyes of 24 patients with an average age of 46 years who underwent two-stage surgery were studied retrospectively. Variables included visual acuity, refractive status, specular microscopy before and after IOL implantation (6 months), and the occurrence of graft failure. Astigmatism was corrected by suture removal and relaxing incision. The mean follow-up after IOL placement was 16 months. RESULTS: Unaided visual acuity was 20/100 or better in 22 (83%) eyes after the second procedure. Twenty-one (81%) eyes had an aided visual acuity of at least 20/40. The mean refractive and absolute errors were -1.49 +/- 1.39 diopters (D) and 1.55 +/- 1.30 D, respectively. The mean keratometric and refractive cylinders were 3.50 D and 2.26 D, respectively. Nineteen (73%) eyes had a spherical equivalent refraction within 2 D of emmetropia. Anisometropia (> or =3 D) occurred in four (15%) eyes. The endothelial cell density, the coefficient of variation, and the percentage of hexagonal cells documented by specular microscopy were not significantly different before and after IOL implantation. Complications included three rejections, two cases of herpetic recurrence, and one late decompensation. Two graft failures (8%) occurred after secondary surgery. CONCLUSION: The two-stage surgery is a safe and effective modality. PMID- 11473157 TI - Corneoscleral rim cultures: lack of utility and implications for clinical decision-making and infection prevention in the care of patients undergoing corneal transplantation. AB - PURPOSE: To assess the utility of donor corneoscleral rim cultures. METHODS: A retrospective review of the culture results of 774 corneoscleral rims that remained after trephination of corneas for transplantation into patients at our academic medical center between January 1992 and November 1997. RESULTS: Forty one (5.3%) corneoscleral rim cultures yielded microorganisms, mostly coagulase negative staphylococci. Two patients developed endophthalmitis (one with Staphylococcus aureus and one with Pseudomonas aeruginosa) within 3 months after transplantation; each had a negative corneoscleral rim culture and neither patient's infection was temporally related to the transplant procedure. CONCLUSIONS: Preoperative donor corneoscleral rim cultures are unreliable predictors of endophthalmitis complicating corneal transplantation and, therefore, are not useful in the clinical management of patients having corneal transplants. Moreover, the discrepancy between the results of corneoscleral rim cultures and subsequent endophthalmitis renders them invalid as a quality assurance procedure. Instead, for patients with suspected endophthalmitis after corneal transplantation, we recommend that corneal surgeons select antimicrobial therapy based on current guidelines and the results of directed sampling. Furthermore, eye banks should prospectively track recipients who develop clinical endophthalmitis, immediately notify the corneal surgeon who transplanted the matched cornea of that used for the index case, and, in selected situations, attempt to identify a possible source of contamination. PMID- 11473158 TI - Corneal transplant tolerance of cryopreservation. AB - PURPOSE: To study the outcome of corneal transplants performed with cryopreserved tissue. METHODS: Maisonneuve-Rosemont Hospital medical records of all corneal transplantations performed with cryopreserved tissue by one surgeon (M.L.F.) between March 1978 and April 1991 were reviewed. The Kaufman--Capella cryopreservation technique was used. Corneas were cryopreserved for periods of 3 days to 16.8 years (mean, 4.6 years) before transplantation. RESULTS: We report a mean follow-up of 54 months (range, 2.8--151.3 months). Survival analysis showed the probability of a clear graft to be 76% at 1 year and 73.2% at 2 years. At the time of the last visit, visual acuity was 20/40 or better in 61 eyes (49.2%). The mean postoperative pachometry was 0.58 mm (range, 0.50--0.75 mm). Specular microscopy performed in 57 eyes showed a mean endothelial cell count of 938 cells/mm(2) 55.1 months (range, 2.9--151.3 months) after surgery. For comparison purposes, the outcome of a subgroup of cryopreserved (n = 33) and noncryopreserved (n = 26) corneas transplanted by the same surgeon between April 1986 and April 1990 was studied. CONCLUSION: Despite an increase in the primary failure rate and higher initial endothelial cell loss, cryopreserved transplants are viable. Although we do not recommend cryopreservation of corneas for elective surgery, we consider that cryopreserved corneas can be very useful in emergency situations when tissue availability is a problem. PMID- 11473159 TI - Acute corneal endothelial changes after laser in situ keratomileusis. AB - PURPOSE: To report the acute effects of laser in situ keratomileusis (LASIK) on the corneal endothelium. METHODS: Twenty eyes of 10 consecutive patients (mean age, 38.1 +/- 10.84 years) underwent bilateral simultaneous LASIK for myopic astigmatism (spherical equivalent ranging from -1.75 to -7.13 diopters) without any complications. Each eye was evaluated by slit-lamp biomicroscopy and noncontact specular microscopy preoperatively, within 15 minutes after LASIK and 1 day after surgery. Specular microscopy images were then analyzed to calculate endothelial cell density (ECD), coefficient of variation (CV) of cell size, and percentage of hexagonal cells. RESULTS: All corneas demonstrated marked alterations in endothelial cell morphology by slit-lamp biomicroscopy within 15 minutes after surgery that resolved by the first postoperative day. Central corneal endothelial analysis by noncontact specular microscopy confirmed pleomorphism with definite loss of hexagonality. Mean ECD was calculated to be 2,816.3 +/- 286.02 cells/mm(2) preoperatively, 2,750.85 +/- 327.95 cells/mm(2) on day 0 (p = 0.395), and 2,810.55 +/- 218.48 cells/mm(2) on day 1 (p = 0.461). Mean CV was 32.65 +/- 7.29 preoperatively, 34.4 +/- 6.19 on day 0 (p = 0.412), and 30.9 +/- 5.54 on day 1 (p = 0.067). Mean percentage of hexagonal cells was 63.35 +/- 10.76 preoperatively, 47.55 +/- 9.69 on day 0 (p = 0.000009), and 60 +/- 9.3 on day 1 (p = 0.00003). CONCLUSION: Qualitative and quantitative changes in endothelial cell morphology (i.e., decreased endothelial cell hexagonality) demonstrate that LASIK does induce an acute effect on the corneal endothelium that may represent transient endothelial cell edema. PMID- 11473160 TI - Nineteen years of penetrating keratoplasty in the Hotel-Dieu Hospital in Paris. AB - PURPOSE: To identify changing trends in penetrating keratoplasties (PKs) performed at the Hotel-Dieu Hospital in Paris between January 1980 and December 1999 and to explain the reasons for the changes. METHODS: We retrospectively reviewed 3,736 of the 3,836 PKs performed between January 1, 1980, and December 31, 1999, and classified them into diagnostic categories. RESULTS: The most common indications for PK were keratoconus (28.8%), herpetic infections (10.9%), graft failures (9.9%), aphakic and pseudophakic corneal edema (9.9%), Fuchs' endothelial dystrophy (9.4%), and nonherpetic leucoma (7.7%). Other indications represented 23.4% of the cases. The incidence of aphakic and pseudophakic corneal edema progressively increased between 1980 and 1991, became the most frequent indication in 1991 (21.4%), and then progressively decreased. The annual number of PKs increased between 1980 and 1986, decreased between 1987 and 1997, and increased again after September 1997. The decrease was caused by both a shortage of corneal buttons, and, in 1987, the fear of transmitting diseases through corneal transplantation, particularly human immunodeficiency virus. Beginning in 1992, decreases were also associated with stringent governmental regulations of eye bank tissue. CONCLUSION: Changes in the incidence and management of corneal disorders were the primary factors leading to modifications of grafting until 1987. After 1987, corneal button shortage probably corresponded to the acquired immune deficiency syndrome epidemic. Governmental regulations of eye banking led to a severe corneal button shortage between 1992 and 1997. Despite an increase in the number of PKs performed after 1997, corneal buttons are still preferentially allocated to patients in whom there is a high probability of graft success. PMID- 11473161 TI - Utility of Etest in choosing appropriate agents to treat fungal keratitis. AB - PURPOSE: To evaluate the utility of Etest in choosing the appropriate treatment of fungal keratitis. METHODS: Etest was used to determine the drug sensitivities of isolates from the eyes of three patients with fungal keratitis, and the clinical outcomes of treatment with selected drugs were evaluated. RESULTS: In all cases, drug sensitivity demonstrated by Etest accorded with clinical efficacy of the drugs. CONCLUSION: The results in these cases suggest that evaluating drug sensitivities with Etest is an efficient means of selecting optimal pharmacotherapy for fungal keratitis. PMID- 11473162 TI - Characterization of ocular surface symptoms from optometric practices in North America. AB - PURPOSE: This study characterized ocular symptoms typical of dry eye in an unselected optometric clinical population in the United States and Canada. METHODS: Self-administered dry eye questionnaires, one for non-contact lens wearers (dry eye questionnaire) and one for contact lens wearers (contact lens dry eye questionnaire), were completed at six clinical sites in North America. Both questionnaires included categoric scales to measure the prevalence, frequency, diurnal severity, and intrusiveness of nine ocular surface symptoms. The questionnaires also asked how much these ocular symptoms affected daily activities and contained questions about computer use, medications, and allergies. The examining doctors, who were masked to questionnaire responses, recorded a nondirected dry eye diagnosis for each patient, based on their own diagnostic criteria. RESULTS: The dry eye questionnaires were completed by 1,054 patients. The most common ocular symptom was discomfort, with 64% of non--contact lens wearers and 79% of contact lens wearers reporting the symptom at least infrequently. There was a diurnal increase in the intensity of many symptoms, with symptoms such as discomfort, dryness, and visual changes reported to be more intense in the evening. The 22% percent of non-contact lens wearers and 15% of contact lens wearers diagnosed with dry eye (most in the mild to moderate categories) reported symptoms at a greater frequency than those not diagnosed with dry eye. CONCLUSIONS: Our results show that symptoms of ocular irritation and visual disturbances were relatively common in this unselected clinical population. The intensity of many ocular symptoms increased late in the day, which suggested that environmental factors played a role in the etiology of the symptoms. PMID- 11473163 TI - Epiphora caused by conjunctivochalasis: treatment with transplantation of preserved human amniotic membrane. AB - PURPOSE: To present our experience with the use of preserved human amniotic membrane on patients with epiphora caused by conjunctivochalasis. METHODS: Twelve patients, seven women and five men, ages ranging from 56 to 72 years (mean, 61 years) were referred to our Cornea Service with chronic epiphora. In all patients, no punctal ectropion was present, the lacrimal pathway was patent, and the dye disappearance test was abnormal. All patients had already undergone various therapies including multiple irrigations of the lacrimal system, antibiotic drops, steroid drops, and artificial tear drops. In all patients, conjunctivochalasis, which was not previously diagnosed, was evident on slit-lamp examination. After surgical removal of the excess conjunctiva, preserved human amniotic membrane was placed over and sutured with 10-0 nylon continuous suture to the free conjunctival edges. During the postoperative period, artificial tear drops and steroid/antibiotic drops were applied. RESULTS: Improvement of the epiphora was evident from the first postoperative day. After removal of the suture 10 to 15 days (mean, 12 days) after surgery, no patient complained of epiphora. The dye disappearance test was normal. During the follow-up period, which ranged from 6 to 11 months (mean, 8 months), no patient complained of epiphora and no conjunctivochalasis was detected in the area in which human amniotic membrane was transplanted. CONCLUSION: In our experience, transplantation of preserved human amniotic membrane greatly improved symptoms of epiphora caused by conjunctivochalasis. Continued education of the general ophthalmologists concerning this condition is required. PMID- 11473164 TI - Tear IgA and serum IgG antibodies against Acanthamoeba in patients with Acanthamoeba keratitis. AB - PURPOSE: Exposure to Acanthamoebaspecies appears to be ubiquitous, as 50% to 100% of healthy human subjects display anti-Acanthamoebaantibodies. However, the presence of specific anti-Acanthamoebaantibodies in the serum and tears of patients has not been investigated. The prevalence of serum immunoglobulin G (IgG) and tear IgA against three species of Acanthamoebawas assessed in healthy subjects and patients with Acanthamoebakeratitis. METHODS: The level of specific serum IgG and tear IgA against A. castellanii, A. astronyxis, and A. culbertsoniin the sera of 23 patients and 25 healthy subjects was tested by enzyme-linked immunosorbent assays. Total serum IgM, IgG, and IgA concentrations were measured by nephelometry. Acanthamoebakeratitis was diagnosed clinically and confirmed by in vivo confocal microscopy. In some patients, corneal biopsies were also performed and trophozoites were cultured on lawns of Escherichia colion non nutrient agar. RESULTS: All healthy subjects and patients with Acanthamoebakeratitis had detectable serum IgG antibodies against all Acanthamoebaantigens. However, patients with Acanthamoebakeratitis had significantly higher anti-AcanthamoebaIgG antibody titers than healthy subjects. In contrast, Acanthamoeba-specific tear IgA was significantly lower in patients with Acanthamoebakeratitis in comparison with healthy subjects. Total serum immunoglobulins did not differ significantly between healthy subjects and patients with Acanthamoebakeratitis. CONCLUSIONS: The results suggest that a low level of anti-AcanthamoebaIgA antibody in the tears appears to be associated with Acanthamoebakeratitis. PMID- 11473165 TI - Amniotic membrane, tear film, corneal, and aqueous levels of ofloxacin in rabbit eyes after amniotic membrane transplantation. AB - PURPOSE: We evaluated ocular penetration and drug levels in tears after topical ofloxacin instillation in rabbit eyes with amniotic membrane transplantation (AMT). METHODS: Forty-eight New Zealand White rabbits were used. In the first set of experiments, 24 rabbits (24 eyes) were divided into four groups according to the epithelial removal or AMT. Topical ofloxacin was instilled four times every 15 minutes. One hour after the last eyedrop, the concentration of ofloxacin in the amniotic membrane, cornea, and aqueous humor was evaluated. In the second set of experiments, 24 rabbits were divided into six groups according to AMT (transplantation of lyophilized or fresh amniotic membrane) or duration of application. Ofloxacin ointment or two drops of ofloxacin were applied to the right eye, and then tear samples were collected after 0.5, 1, 2, 4, and 6 hours for the analysis of ofloxacin concentration. RESULTS: Mean ofloxacin concentrations in the cornea and aqueous humor were statistically higher in deepithelialized cornea regardless of AMT (p < 0.05). The mean tear levels of ofloxacin in the AMT groups were statistically higher than those in non-AMT groups (p < 0.05). There was no statistical significance in the tear level of ofloxacin between lyophilized amniotic membrane groups and fresh amniotic membrane groups nor between 1-hour amniotic membrane-attached groups and 6-hour amniotic membrane-attached groups. CONCLUSION: Amniotic membrane transplantation seems to interfere with the ocular penetration of topical ofloxacin in normal rabbit corneas but enhances ofloxacin penetration in corneas with epithelial defects. The ofloxacin level in tears was higher in eyes with AMT up to 1 hour after topical ofloxacin use. Therefore, it seems that amniotic membrane has some potential to act as an effective drug delivery system. PMID- 11473166 TI - Adherence of Acanthamoeba to unworn conventional and disposable soft contact lenses. AB - PURPOSE: To compare the adherence of Acanthamoeba trophozoites and cysts to two types of unworn conventional and disposable contact lenses included in groups 1 and 4 of the Food and Drug Administration and to assess the adherence to the center and edge of the lenses. METHODS: We used 10 conventional and 10 disposable contact lenses belonging to groups 1 and 4 of the Food and Drug Administration. Each lens was divided into four segments. A human corneal isolate of Acanthamoeba suspended in saline (1x 105 amoebae/mL) was used. Lens segments were exposed to Acanthamoeba cysts and trophozoites for 2 hours. After removal, the count was made with an optical microscope at 100x magnification. Three fields at the center and three fields at the edge were read on each segment. RESULTS: Acanthamoeba adhered to both types of contact lenses, with a proportion of trophozoites to cysts between 6:4 and 7:3. There was a significantly higher adherence to disposable (8.14 +/- 3.57 organisms/mm(2)) than to conventional (4.94 +/- 2.33 organisms/mm(2)) (p < 0.0001) contact lenses. We also found that the greatest adherence was to the edge of the conventional (12.05 +/- 4.03 organisms/mm(2)) and disposable (26.10 +/- 7.84 organisms/mm(2) (p < 0.0001) contact lenses. CONCLUSIONS: This study shows that Acanthamoeba has varying adherence affinity for different new hydrogel contact lenses. The affinity for disposable lenses (etafilcon A) is significantly higher than that for conventional lenses (polymacon). This adherence is greater for ionic material and high water content. The use of low water content, nonionic contact lenses may reduce the adherence of Acanthamoeba. Finally, the edge may play an important role as an Acanthamoeba vector. PMID- 11473167 TI - Saxitoxin: an anesthetic of the deepithelialized rabbit cornea. AB - PURPOSE: The aim of the current study was to define the efficacy of saxitoxin as a corneal anesthetic in rabbits after mechanical corneal abrasion and photorefractive keratectomy (PRK). METHODS: Twelve Dutch belted rabbits were given a single 1.2-microg dose of saxitoxin or vehicle after mechanical abrasion of the cornea. Corneal sensation was evaluated hourly for 6 hours. A second group of 12 Dutch belted rabbits was given a 1.2-microg dose of saxitoxin or vehicle every 5 hours for 30 hours after PRK. Corneal sensation was evaluated after 5, 10, 15, 20, 25, and 30 hours. Pachometry was performed before PRK and again after the epithelial defects had healed. The rate of epithelial defect closure was assessed by measuring the epithelial defect size 25, 42, 65, 88, and 113 hours after PRK. RESULTS: A dose of 1.2 microg of saxitoxin given every 5 hours produced continuous corneal anesthesia after PRK. There was no difference in the rate of wound healing between eyes treated with saxitoxin and vehicle. There was no difference in the degree of wound healing, as measured by pachometry, between eyes treated with saxitoxin and vehicle. There were no apparent ocular or systemic toxic effects from saxitoxin administration. CONCLUSION: At a dose of 1.2 microg, saxitoxin is a safe, effective, long-acting corneal anesthetic in rabbits after PRK. PMID- 11473168 TI - Corneal virulence of LasA protease--deficient Pseudomonas aeruginosa PAO1. AB - PURPOSE: Pseudomonas aeruginosa PAO1 deficient in LasA protease was reported to be ocularly avirulent. However, the avirulence of this mutant could not attributed to the loss of LasA protease. The purpose of this study was to define the mechanism for such a mutant's inability to cause corneal disease. METHODS: A LasA protease--deficient mutant of P. aeruginosa PAO1 was constructed by allelic exchange. Virulence of this mutant in mouse and rabbit models of keratitis was assessed by scoring for ocular disease and quantitating viable bacteria from infected corneas. Adherence to scarified mouse corneal tissue was determined with an organ culture assay. RESULTS: In the mouse eye, the LasA protease--deficient mutant was not virulent, despite being as adherent as its parent strain. Virulence of the mutant was also significantly reduced in the rabbit eye. Complementation with lasA did not restore virulence in either model of infection. Neither the mutant nor the mutant complemented with lasA grew well in ocular tissue. An analysis of the mutant showed that it was auxotrophic for leucine. CONCLUSION: These data show that the mutant's avirulence in the eye is caused by poor growth in the ocular environment and not the loss of a functional lasA gene. PMID- 11473169 TI - Influence of dacron tissue thickness on the performance of the Pintucci biointegrable keratoprosthesis: an in vitro and in vivo study. AB - PURPOSE: In 1979 Pintucci developed a biointegrable keratoprosthesis with polymethylmethacrylate optical cylinder integrated with a Dacron tissue colonizable supporting element to avoid the complications caused by the interaction between the haptic element and the eye. The purpose of this article is to compare the colonization of three Dacron fabrics (thicknesses of 0.25 mm, 0.6 mm, and 1.4 mm) in vitro and in vivo to optimize the device performance. METHODS: In vitro three different Dacron fabrics were cultured for 3 days with 3.5 x 10(5) human fetal lung fibroblasts and observed with a scanning electron microscope. In vivo three different Dacron fabrics were implanted on the sclera near the superior rectus insertion in the right eye of six albino rabbits and were observed after 4 days with light and scanning electron microscopy. RESULTS: In the in vitro experiments, the cells were preserved and their structure was found to be normal. The 0.25-mm thick fabric was coated only on the surface, and the other fabrics were colonized in three dimensions. In the in vivo experiments, the 0.25-mm thick fabric appeared coated only on its surface. The other fabrics were three-dimensionally colonized and the Dacron filaments appeared embedded in neovascularized connective tissue with minimal foreign body reaction. The 1.4-mm thick colonized fabric showed a substantial loss of pliability. CONCLUSION: Given that the 0.25-mm thick fabric was coated only by connective tissue, that the 0.6 mm and 1.4-mm thick fabrics were perfectly colonized, and that the 1.4-mm thick fabrics showed a substantial loss of pliability, the 0.6-mm thick fabric haptic part of the Pintucci keratoprosthesis is preferred. For 19 years, the 0.6-mm Dacron fabric Pintucci keratoprosthesis was implanted in 159 eyes with good results, overcoming the apparently inseparable difficulties represented by mechanical anchorage and biointegrability of a keratoprosthesis. PMID- 11473170 TI - Surface ultrastructure of collagen fibrils and their association with proteoglycans in human cornea and sclera by atomic force microscopy and energy filtering transmission electron microscopy. AB - PURPOSE: We aimed to investigate the possible association of proteoglycans with D periodic collagen fibrils in the human cornea and sclera, using energy-filtering transmission electron microscopy (EF-TEM) and atomic force microscopy (AFM). METHODS: Human cornea and sclera were digested with keratanase to eliminate keratan sulfate proteoglycans (KSPGs). For EF-TEM observation, surface proteoglycans were detected by cupromeronic blue (CB) staining. For AFM observation, cornea and sclera were treated with sodium hydroxide before and after keratanase digestion, and the surface topology of collagen fibrils was analyzed. RESULTS: With CB staining, numerous CB-positive short filaments of surface proteoglycans (proteoglycan filaments) were observed in the interfibrillar spaces of cornea and sclera associated with collagen fibrils. AFM imaging showed that the depth and periodicity of D-periodic collagen fibrils in keratanase-treated corneal collagens were deeper and more regular than in untreated ones. Moreover, the depth and periodicity of keratanase-untreated corneal collagens were shallow and irregular in comparison with keratanase untreated scleral collagens. On the other hand, there was no difference in depth or regularity between keratanase-treated and -untreated scleral collagen fibrils. Using AFM imaging, additional thin grooves sub-bands were detected on the surface of keratanase-treated corneal collagen fibrils. The grooves were not detected in keratanase-untreated collagen fibrils nor in scleral collagen fibrils with or without keratanase digestion. Comparing densitometry waves, the grooves of D periodic corneal collagen sub-bands corresponded to a and c bands. CONCLUSION: Using AFM and EF-TEM to study corneal and scleral collagen fibrils and their association with proteoglycans, we conclude that KSPG is found in ample amounts in the human cornea in comparison with sclera. Moreover, we topologically detected KSPG attached to a and c bands of collagen fibrils. PMID- 11473171 TI - Short-term treatment With a potent topical corticosteroid of an acute ocular adenoviral infection in the New Zealand white rabbit. AB - PURPOSE: To determine the effect of short-term topical therapy with 1% prednisolone acetate (PA) on normal immune adenoviral clearance in the rabbit ocular model. METHODS: Thirty rabbits were topically inoculated in both eyes with adenovirus type 5 (Ad5). On day 1, the rabbits were divided into three topical treatment groups: 1% PA four times daily for 3 days, 1% PA four times daily for 5 days, control (artificial tears) four times daily for 5 days. Eyes were cultured for virus on days 0, 1, 3, 4, 5, 7, 9, 11, and 14. RESULTS: Compared with the control group, treatment with 1% PA for 3 or 5 days significantly increased the total and daily number of Ad5-positive cultures from days 7 to 14, prolonged the duration of Ad5 shedding, and increased the mean combined Ad5 titer from days 1 to 5. In addition, treatment with 1% PA for 5 days increased the mean combined Ad5 titer from days 7 to 14. CONCLUSION: Treatment of an ocular adenoviral infection with 1% PA for as little as four times daily for 3 days significantly enhanced adenoviral replication compared with the control group. Short-term corticosteroid treatment of acute adenoviral ocular infections with 1% PA should be used judiciously. PMID- 11473172 TI - Cat's tooth keratitis: human corneal infection with Capnocytophaga canimorsus. AB - PURPOSE: To describe a patient with keratitis caused by Capnocytophaga canimorsus after traumatic injury with a cat's tooth. METHODS: Case report and review of the literature. RESULTS: Sixteen months after onset of infection, the patient regained 20/30 vision in the affected eye. However, he required 7 months of antibiotic therapy to eliminate the infection. CONCLUSION: C. canimorsus keratitis may follow penetrating corneal injury and wound contamination with cat saliva and may be recalcitrant to topical antibiotic therapy. PMID- 11473173 TI - Bilateral corneal involvement in epidermolysis bullosa acquisita. AB - PURPOSE: To report clinical and laboratory findings of bilateral corneal involvement in a patient with epidermolysis bullosa acquisita. METHODS: A 25-year old man with a history of progressive and painless loss of vision in both eyes presented to our service with bilateral corneal involvement: peripheral corneal perforation in one eye and advanced corneal thinning in the other eye. There was concomitant dermatologic bullous disease. Clinical and laboratory exams were analyzed. RESULTS: The patient was diagnosed as having epidermolysis bullosa acquisita. Therapeutic corneal patch graft and conjunctival resection with cryotherapy were done, with satisfactory results. CONCLUSION: Bilateral corneal involvement in epidermolysis bullosa acquisita is described. To the best of our knowledge, this is the first description of such a case. Surgical management of the ocular findings associated with systemic therapy with colchicine seems to be a good therapeutic option in the management of this defying disease. PMID- 11473174 TI - Pseudokeratoconus caused by rosacea. AB - PURPOSE: A case of inferior corneal thinning and high astigmatism with features of keratoconus in a patient with long-standing ocular rosacea is described. METHODS: Axial curvature mapping was performed with the Tomey TMS-1 videokeratoscopy instrument and corneal thickness mapping was performed with the Orbscan Corneal Topography System (CTS). Tear clearance was assessed by measuring the concentration of fluorescein in the tear fluid with a fluorometer. RESULTS: There were inferior corneal thinning and opacification in both eyes. Tear fluorescein clearance was markedly delayed in the right eye. There was asymmetric inferior corneal steepening in both eyes with I-S values of 1 in the right eye and 5.9 in the left eye. There were 5.9 diopters of astigmatism at 85 degrees in the right eye and 7.3 diopters of astigmatism at 73 degrees in the left eye. Corneal pachymetry mapping with the Orbscan CTS showed a normal central corneal thickness and maximal thinning in the inferotemporal periphery of the right cornea and inferonasal periphery of the left cornea. CONCLUSION: Chronic ocular rosacea can produce inferior corneal thinning and high astigmatism with some features of keratoconus. The inferior pattern of thinning in rosacea may be related to chronic exposure of the inferior cornea to inflammatory and matrix degrading factors in the inferior tear meniscus. PMID- 11473175 TI - Consecutive Descemet membrane detachment after successive phacoemulsification. AB - PURPOSE: To report a patient with consecutive Descemet's membrane (DM) detachments after successive phacoemulsification, review other reported patients with bilateral DM detachments, and explore the possibility of anatomic predisposition to DM detachment in some patients. METHODS: Our patient's course was reviewed along with the reported experience with three other patients with bilateral DM detachments. RESULTS: No clear underlying etiology of DM detachment was found in our patient or any of the other three reported patients reviewed. CONCLUSIONS: Some patients may be anatomically predisposed to DM detachment possibly because of an abnormality in the fibrillary stromal attachment to DM. Early postoperative surgical intervention often leads to satisfactory visual results. PMID- 11473176 TI - The molecular genetics of Bowman's layer dystrophies. PMID- 11473178 TI - Lateralization of brain activation to imagination and smell of odors using functional magnetic resonance imaging (fMRI): left hemispheric localization of pleasant and right hemispheric localization of unpleasant odors. AB - PURPOSE: Our goal was to use functional MRI (fMRI) of brain to reveal activation in each cerebral hemisphere in response to imagination and smell of odors. METHOD: FMRI brain scans were obtained in 24 normal subjects using multislice fast low angle shot (FLASH) MRI in response to imagination of banana and peppermint odors and in response to smell of corresponding odors of amyl acetate and menthone, respectively, and of pyridine. Three coronal sections selected from anterior to posterior brain regions were used. Similar studies were obtained in two patients with hyposmia using FLASH MRI and in one patient with hyposmia using echo planar imaging (EPI) both before and after theophylline treatment that returned smell function to or toward normal in each patient and in two patients with birhinal phantosmia (persistent foul odor) and global phantogeusia (persistent foul taste) with FLASH and EPI fMRI before and after treatment with neuroleptic drugs that inhibited their phantosmia and phantogeusia. Activation images were derived using correlation analysis. Ratios of hemispheric areas of brain activation to total hemispheric brain areas were calculated for FLASH fMRI, and numerical counts of pixel clusters in each hemisphere were made for EPI studies. Total pixel cluster counts in localized regions of each hemispheric section were also obtained. RESULTS: In normal subjects, activation generally occurred in left (L) > right (R) brain hemisphere in response to banana and peppermint odor imagination and to smell of corresponding odors of amyl acetate and menthone. Whereas there were no overall hemispheric differences for pyridine odor, activation in men was R > L hemisphere. Although absolute activation in both L and R hemispheres in response to banana odor imagination and amyl acetate smell was men > women, the ratio of L to R activation was women > men. In hyposmic patients studied by FLASH fMRI, activation to banana odor imagination and amyl acetate smell was L > R hemisphere both before and after theophylline treatment. In the hyposmic patient studied with EPI before theophylline treatment, activation to banana and peppermint odor imagination and to amyl acetate, menthone, and pyridine smell was R > L hemisphere; after theophylline treatment restored normal smell function, activation shifted completely with banana and peppermint odor imagination and amyl acetate and menthone smell to L > R hemisphere, consistent with responses in normal subjects. However, this shift also occurred for pyridine smell, which is opposite to responses in normal control subjects. In patients with phantosmia and phantogeusia, activation to phantosmia and phantogeusia before treatment was R > L hemisphere; after treatment inhibited phantosmia and phantogeusia, activation shifted with a slight L > R hemispheric lateralization. Localization of all lateralized responses indicated that anterior frontal and temporal cortices were brain regions most involved with imagination and smell of odors and with phantosmia and phantogeusia presence. CONCLUSION: Imagination and smell of odors perceived as pleasant generally activated the dominant or L > R brain hemisphere. Smell of odors perceived as unpleasant and unpleasant phantosmia and phantogeusia generally activated the contralateral or R > L brain hemisphere. With remission of phantosmia and phantogeusia, hemispheric activation was not only inhibited, but also there was a slight shift to L > R hemispheric predominance. Predominant L > R hemispheric differences in brain activation in normal subjects occurred in the order amyl acetate > menthone > pyridine, consistent with the hypothesis that pleasant odors are more appreciated in L hemisphere and unpleasant odors more in R hemisphere. Anterior frontal and temporal cortex regions previously found activated by imagination and smell of odors and phantosmia and phantogeusia perception accounted for most hemispheric differences. PMID- 11473179 TI - High b-value diffusion-weighted MRI of normal brain. AB - PURPOSE: As MR scanner hardware has improved, allowing for increased gradient strengths, we are able to generate higher b values for diffusion-weighted (DW) imaging. Our purpose was to evaluate the appearance of the normal brain on DW MR images as the diffusion gradient strength ("b value") is increased from 1,000 to 3,000 s/mm2. METHOD: Three sets of echo planar images were acquired at 1.5 T in 25 normal subjects (mean age 61 years) using progressively increasing strengths of a diffusion-sensitizing gradient (corresponding to b values of 0, 1,000, and 3,000 s/mm2). All other imaging parameters remained constant. Qualitative assessments of trace images were performed by two neuroradiologists, supplemented by quantitative measures of MR signal and noise in eight different anatomic regions. RESULTS: As gradient strength increased from b = 1,000 to 3,000, both gray and white matter structures diminished in signal as expected based on their relative diffusion coefficients [calculated average apparent diffusion coefficient (ADC) values: gray matter = 8.5 x 10(-4) mm2/s, white matter = 7.5 x 10(-4) mm2/s]. The signal-to-noise ratios for the b = 1,000 images were approximately 2.2 times higher than for the b = 3,000 images (p < 0.0001). As the strength of the diffusion-sensitizing gradient increased, white matter became progressively hyperintense to gray matter. Relative to the thalamus, for example, the average MR signal intensity of white matter structures increased by an average of 27.5%, with the densely packed white matter tracts (e.g., middle cerebellar peduncle, tegmentum, and internal capsule) increasing the most. CONCLUSION: Brain DW images obtained at b = 3,000 appear significantly different from those obtained at b = 1,000, reflecting expected loss of signal from all areas of brain in proportion to their ADC values. Consequently, when all other imaging parameters are held constant, b = 3,000 DW images appear significantly noisier than b = 1,000 images, and white matter tracts are significantly more hyperintense than gray matter structures. PMID- 11473180 TI - CT angiography in the rapid triage of patients with hyperacute stroke to intraarterial thrombolysis: accuracy in the detection of large vessel thrombus. AB - PURPOSE: The purpose of this work was to evaluate the accuracy of CT angiography (CTA) for the detection of large vessel intracranial thrombus in clinically suspected hyperacute (<6 h) stroke patients. METHOD: Forty-four consecutive intraarterial thrombolysis candidates underwent noncontrast CT followed immediately by CTA. Axial source and two-dimensional collapsed maximum intensity projection reformatted CTA images were rated for the presence or absence of large vessel occlusion. Five hundred seventy-two circle-of-Willis vessels were reviewed; arteriographic correlation was available for 224 of these. RESULTS: Sensitivity and specificity for the detection of large vessel occlusion were 98.4 and 98.1%; accuracy, calculated using receiver operating characteristic analysis, was 99%. Mean time for acquisition, reconstruction, and analysis of CTA images was approximately 15 min. CONCLUSION: CTA is highly accurate for the detection and exclusion of large vessel intracranial occlusion and may therefore be valuable in the rapid triage of hyperacute stroke patients to intraarterial thrombolytic treatment. PMID- 11473181 TI - Mapping therapeutic response in a patient with malignant glioma. AB - Short-interval scanning of patients offers a detailed understanding of the natural progression of tumor tissue, as revealed through imaging markers such as contrast enhancement and edema, prior to therapy. Following treatment, short interval scanning can also provide evidence of attenuation of growth rates. We present a longitudinal imaging study of a patient with glioblastoma multiforme (GBM) scanned 15 times in 104 days on a 3 T MR scanner. Images were analyzed independently by two automated algorithms capable of creating detailed maps of tumor changes as well as volumetric analysis. The algorithms, a nearest-neighbor based tissue segmentation and a surface-modeling algorithm, tracked the patient's response to temozolomide, showing an attenuation of growth. The need for surrogate imaging end-points, of which growth rates are an example, is discussed. Further, the strengths of these algorithms, the insight gained by short-interval scanning, and the need for a better understanding of imaging markers are also described. PMID- 11473182 TI - Diffusion property in a hamartomatous lesion of neurofibromatosis type 1. AB - Though diffusion-weighted MRI has been applied to various intracranial lesions, few reports had been presented about cerebral hamartomatous lesions in patients with neurofibromatosis type 1 (NF1). In this study, we report the interval changes of apparent diffusion coefficient (ADC) in a presumed hamartomatous lesion. In our case, the ADC increased slightly over a 3 year period. This diffusion property may provide specific insight into the etiology of cerebral hamartomatous lesions observed in NF1. PMID- 11473183 TI - Imaging studies in the diagnostic workup of neonatal nasal obstruction. AB - Twelve neonates presenting with nasal obstruction after birth were evaluated by imaging studies for diagnostic reasons. Four groups were recognized: Group I: choanal atresia (n = 5) and choanal stenosis (n = 1); Group II: congenital nasal pyriform aperture stenosis (CNPAS) (n = 3) and holoprosencephaly (n = 1); Group III: nasolacrimal duct mucocele (n = 1); Group IV: nasal hypoplasia (n = 1). Associated anomalies were found in eight patients. Four patients with choanal atresia showed manifestations of the CHARGE (coloboma, congenital heart defect, atretic choanae, retarded physical and neuromotor development associated with central nervous system anomalies, genital hypoplasia, and ear anomaly and/or deafness) association. In the fifth patient with choanal atresia, the diagnosis of amnion disruption sequence was made. One patient with CNPAS had a solitary maxillary central incisor (SMCI), a mild form of holoprosencephaly. Besides proboscis and synophthalmos, SMCI was also present in the holoprosencephaly case. The patient with severe nasal hypoplasia had warfarin embryopathy. This review emphasizes the need for performing imaging studies in the diagnostic workup of neonates born with nasal obstruction. PMID- 11473184 TI - Cardiac phase contrast gradient echo MRI: characterization of abnormal left ventricular wall motion in patients with ischemic heart disease. AB - PURPOSE: In our patient study, we examined the clinical usefulness of phase contrast velocity mapping for the detection and characterization of localized abnormalities of left ventricular motion. MATERIALS AND METHODS: Velocity encoding is based on the fact that motion in the presence of a magnetic field gradient causes a change of the phase of the MRI signal that is proportional to the velocity of tissue motion. Left ventricular motion was characterized by parameters describing rotation and contraction/dilatation, respectively. We examined 34 patients with localized abnormalities of left ventricular motion due to ischemic heart disease. RESULTS: Three patients could not be sufficiently evaluated due to technical problems including varying positions of the heart during successive breathhold periods. In 27 of the remaining 31 patients, MRI could demonstrate abnormal radial velocities that corresponded fully or partly with perfusion deficits in single photon emission computed tomography or positron emission tomography. The abnormalities were most pronounced in early diastole. Rotational velocities did not show any regional changes. CONCLUSION: Our study showed that our technique is suitable for the detection and characterization of localized abnormalities of left ventricular motion in patients with ischemic heart disease. PMID- 11473185 TI - Serial motion assessment by reference tracking (SMART): application to detection of local functional impact of chronic myocardial ischemia. AB - PURPOSE: To compare measurements of wall motion and thickening with and without correcting for cardiac twisting and shortening. METHOD: Inversion recovery Gd DPTA perfusion and cine motion MRI were performed on 12 pigs with chronic ischemia induced by ameroid occluder. Analyses were based on conventional fixed plane imaging and serial motion assessment by reference tracking (SMART). RESULTS: Normal motion was 31.3 +/- 1.9%, and normal wall thickening was 41.4 +/- 2.2%. At the maximum perfusion defect, SMART wall motion was 10.5 +/- 2.4% and fixed wall motion was 20.6 +/- 1.7% (p < 0.004), SMART wall thickening was 20.1 +/- 4.4%, and fixed wall thickening was 32 +/- 1.9% (p < 0.03). CONCLUSION: SMART measurements of wall thickening and motion detect much smaller thickening and motion in ischemic myocardium than fixed radial metrics. SMART data, covering the entire heart, should prove twice as sensitive to abnormalities in motion and thickening, such as any produced by ischemic heart disease or improved by treatment. PMID- 11473186 TI - CT findings following the cabrol composite graft procedure. AB - PURPOSE: Insertion of a composite graft and reimplantation of the coronary arteries through an intermediate Dacron tube (Cabrol composite graft procedure) has been used to treat ascending aortic aneurysms and dissections. The CT findings after the Cabrol composite graft procedure have not been previously described. METHOD: Retrospective review of 12 postoperative CT and CT angiography (CTA) studies both in the immediate postoperative period as well as during long term follow-up was conducted. RESULTS: The Cabrol composite graft procedure produces typical CT findings consisting of a coronary conduit separate from the aortic graft. The presence of perigraft flow can be normal or abnormal depending on the time point of its occurrence and the extent of its hemodynamic consequences. CONCLUSION: Knowledge of the typical CT and CTA findings following a Cabrol composite graft procedure is essential for the correct interpretation of these studies. PMID- 11473187 TI - Overlapping cross-sections significantly improve the reproducibility of coronary calcium measurements by electron beam tomography: a phantom study. AB - PURPOSE: We conducted phantom studies to investigate whether overlapping cross sections and volumetric scoring would significantly improve interscan reproducibility of electron beam tomography (EBT) for coronary artery calcium quantification. METHOD: Fifteen phantoms simulating various amounts of coronary calcification were scanned in five different positions with a slice thickness of 3.0 mm and a table feed of 3.0, 2.5, and 2.0 mm. For the conventional "Agatston score" and a "volume score" (total volume of calcified lesions), interscan variabilities were compared between the three image acquisition protocols. RESULTS: Agatston score variability was significantly lower for the 2.0 mm table feed than for the 3.0 or 2.5 mm table feed (3.0 mm: 22.9 +/- 10.3%; 2.5 mm: 13.6 +/- 8.2%; 2.0 mm: 8.9 +/- 5.5%). Volume score variability was significantly lower for 2.5 and 2.0 mm table feed than for 3.0 mm table feed (3.0 mm: 21.7 +/- 11.0%; 2.5 mm: 10.9 +/- 5.9%; 2.0 mm: 9.8 +/- 5.9%). CONCLUSION: Overlapping cross sections, especially in combination with volumetric scoring, significantly improved interscan reproducibility of EBT calcium quantification in a phantom study. PMID- 11473188 TI - Postoperative rupture of the right ventricular free wall with transthoracic leak: multidetector CT findings. AB - Right ventricular free-wall rupture may complicate sternal-splitting operations but may be clinically occult. Awareness of this entity and the CT imaging features may help avert a potentially catastrophic outcome. PMID- 11473189 TI - CT and MR findings in a calcified myocardial tuberculoma of the left ventricle. AB - Tuberculosis can involve a multitude of organ tissues but generally affects the respiratory tract. Heart tuberculosis is rare, and the pericardium is the most common location reported in the literature, accounting for 0.5% of extrapulmonary tuberculosis. Isolated myocardial tuberculosis is a very unusual occurrence, with only a few case reports in the literature, usually diagnosed at necropsy. Findings of myocardial tuberculoma on cross-sectional images previously have not been reported. We present computed tomography and magnetic resonance imaging findings of an isolated calcified myocardial tuberculoma. PMID- 11473190 TI - Stent grafting of abdominal aortic aneurysms: pre-and postoperative evaluation with multislice helical CT. AB - Endovascular stent grafting of abdominal aortic aneurysms is a new technique that may replace open surgery in selected cases. Pre-and postoperative angiography can be replaced by helical CT. This pictorial essay describes and illustrates the use of multislice helical CT where maximum intensity projection and multiplanar reformats play a central role in the evaluation. PMID- 11473191 TI - Automated segmentation and visualization of the pulmonary vascular tree in spiral CT angiography: an anatomy-oriented approach based on three-dimensional image analysis. AB - A new method for automated segmentation of the pulmonary vascular tree in spiral CT angiography was developed based on 3D image analysis techniques and anatomic knowledge. For efficient and effective segmentation, an anatomy-oriented approach was introduced, in which several anatomic structures are segmented sequentially and the properties of each segmented structure are used for the next step of segmentation and for validation of intermediate results. By use of clinical data of 12 patients, parameters for segmentation were analyzed and optimized. The effectiveness of the segmentation method was evaluated through the visual assessment by comparison between images of the segmentation results by volume rendering and images of maximum intensity projection of the original volume data. PMID- 11473192 TI - Three-dimensional volume-rendering CT angiography in vasculitis: spectrum of disease and clinical utility. AB - Spiral computed tomographic angiography (CTA) coupled with three-dimensional volume-rendering image processing is a less invasive alternative to conventional catheter angiography. The technique has been used successfully in a variety of vascular diseases. In this pictorial essay, we review the CTA findings in selected cases of vasculitis. Technical considerations and the potential clinical value of this method are discussed. PMID- 11473193 TI - CT differentiation of mesenteric ischemia due to vasculitis and thromboembolic disease. AB - PURPOSE: The purpose of this work was to differentiate CT findings of acute mesenteric ischemia due to vasculitis (MV) or thromboembolism (MTE). METHOD: CT scans of 69 patients with mesenteric ischemia caused by MV (n = 37) or MTE (n = 32) were analyzed. After dividing the patients into groups with and without MV, we compared them with regard to gastrointestinal tract involvement patterns, mesenteric changes, and presence or absence of vascular thrombosis, atherosclerosis, other organ changes, and ascites. RESULTS: Duodenum was involved only in the MV group (30%). The MV group had preferential involvement of the small intestine (89%) to large intestine (51%), whereas the MTE group showed even distribution. Multisegmental bowel involvement was more common in the MV group (86%) than in the MTE group (44%), especially in both jejunum and ileum and both small and large intestine. The MV group showed prominent involvement of the superior mesenteric vessel territory, although the MTE group showed even distribution. Splenomegaly and hydronephrosis were more frequently seen in the MV group and vascular thrombosis and atherosclerosis in the MTE group. CONCLUSION: Although considerable overlap was noted, CT is useful in differentiating MV from MTE. PMID- 11473194 TI - Crohn disease of the small bowel: spiral CT evaluation after oral hyperhydration with isotonic solution. AB - PURPOSE: Our aim was to evaluate the accuracy of spiral CT study of small-bowel Crohn disease with use of oral hyperhydration with isoosmotic solution. METHOD: We prospectively analyzed 33 consecutive patients and 10 control subjects with spiral CT after oral administration of 2,000 ml of polyethylene glycol electrolyte-balanced solution. The CT diagnoses were compared with the results of conventional radiologic oral barium examination (33 cases), ileum colonoscopy (8 cases), and surgery (4 cases). RESULTS: The final diagnoses were Crohn disease (14 cases), no small-bowel disease (16 cases), cancer of ileocecal valve (1 case), carcinosis of mesenteric root (1 case), and intestinal lymphangiectasia (1 case). In the control group, no abnormalities of the small bowel were found. The sensitivity of spiral CT was 85.7%, specificity 100%, positive predictive value 100%, negative predictive value 90%, and diagnostic accuracy 93.9%. CONCLUSION: Our method allowed adequate distension of the small bowel for spiral CT studies, thus resulting in a safe and effective alternative to small-bowel spiral CT enema, which can be used in patients that refuse the nasojejunal balloon catheter. PMID- 11473195 TI - MRI of ganglioneuroma: histologic correlation study. AB - PURPOSE: The purpose of this study was to evaluate the MR findings of ganglioneuroma and to correlate imaging and histologic features. METHOD: Conventional SE and contrast-enhanced dynamic MRI was used to examine 10 patients with pathologically confirmed ganglioneuroma. The morphologic features, signal intensity, and dynamic enhancement pattern of the tumors were retrospectively analyzed and correlated with histologic features. RESULTS: Capsules were present in all tumors histologically and were also detected in five tumors on postcontrast T1-weighted images. A whorled appearance corresponding to interlacing bundles of Schwann cells and collagen fibers on histologic specimens was visualized in five tumors on T1-and/or T2-weighted images. Tumors with markedly high signal intensity on T2-weighted images consisted histologically of a large amount of myxoid stroma and relatively few cellular and fibrous components. Tumors with intermediate to high signal intensity consisted of numerous cellular and fibrous components and little myxoid stroma. Early enhancement of tumors was usually lacking in dynamic MR studies; however enhancement gradually increased. CONCLUSION: The results suggest that the MR features of ganglioneuroma are well correlated with histologic findings. PMID- 11473196 TI - Hepatocellular carcinoma after systemic chemotherapy: gadolinium-enhanced mr measurement of necrosis by volume histogram. AB - As a preliminary study, we measured the necrosis of advanced hepatocellular carcinoma (HCC) by volume histogram after systemic chemotherapy and correlated it with clinical data. Five patients with advanced HCC secondary to chronic hepatitis and cirrhosis underwent pretreatment and posttreatment MR examination on a 1.5 T MR scanner following systemic chemotherapy. MR sequences included dynamic enhanced fast spoiled gradient echo 3D images. Clinical response to chemotherapy, as determined by MR images, was measured as changes of both the total tumor volume and the percent of tumor necrosis by volume histogram algorithm. Four of five patients had clinical improvement. Three of these patients had no or minimal change of tumor volume; however, there was an increase in tumor necrosis in follow-up MR image. One patient of five with no change in tumor necrosis had no response and died at 3 months. Serial MR images showed increased irregular necrosis of advanced HCC after systemic chemotherapy, but stable volume, in patients who responded clinically to systemic chemotherapy. PMID- 11473197 TI - Automated segmentation of colonic walls for computerized detection of polyps in CT colonography. AB - PURPOSE: A new method for fully automated segmentation of the colonic walls in volumetric CT data was developed for limitation of the search space in computerized detection of polyps. METHOD: For reliable segmentation, an anatomy oriented approach was used, in which several anatomical structures are segmented in addition to the colon for utilization of their properties. RESULTS: The segmentation method was validated by use of 14 data sets, consisting of cases positive for colonic polyps. We found that the segmented colonic walls included all of the polyps. A subjective rating of the results was performed based on several criteria for visualization of anatomic detail of the colonic wall and mucosal surface. Except for a few cases in which insufflation of the colon was insufficient, all of the results included >95% of the colonic walls. CONCLUSION: This method for colonic wall segmentation is reliable and the segmentation results are applicable in both visualization of the colon and computer-aided diagnosis in the detection of polyps in CT colonography. PMID- 11473198 TI - Knowledge-based segmentation of pediatric kidneys in CT for measurement of parenchymal volume. AB - PURPOSE: The purpose of this work was to develop an automated method for segmenting pediatric kidneys in helical CT images and measuring their volume. METHOD: An automated system was developed to segment the kidneys. Parametric features of anatomic structures were used to guide segmentation and labeling of image regions. Kidney volumes were calculated by summing included voxels. For validation, the kidney volumes of four swine were calculated using our approach and compared with the "true" volumes measured after harvesting the kidneys. Automated volume calculations were also performed in a cohort of nine children. RESULTS: The mean difference between the calculated and measured values in the swine kidneys was 1.38 ml. For the pediatric cases, calculated volumes ranged from 41.7 to 252.1 ml/kidney, and the mean ratio of right to left kidney volume was 0.96. CONCLUSION: These results demonstrate the accuracy of a volumetric technique that may in the future provide an objective assessment of renal damage. PMID- 11473199 TI - 3,4-Methylenedioxymethamphetamine (ecstasy)-induced hepatotoxicity: multidetector CT and pathology findings. AB - 3,4-methylenedioxymethamphetamine ("Ecstasy") abuse may cause severe hepatotoxicity. The computed tomography and pathologic findings of such a case are described. PMID- 11473200 TI - A case of abnormal pancreaticobiliary junction evidenced by 3D drip infusion cholangiography CT. AB - We report a case of abnormal pancreaticobiliary junction (APBJ), in which three dimensional (3D) drip infusion cholangiography (DIC) computed tomography (CT) was useful to make the diagnosis. The 3D DIC CT clearly demonstrated reflux of the contrast material into the main pancreatic duct as well as a long common channel, dilated biliary tree, and duodenal lumen. Thereby, we could diagnose APBJ. We believe that 3D DIC CT is a valuable method in the evaluation of patients with suspected APBJ. PMID- 11473201 TI - Sectional neuroanatomy of the lower limb I: lower back and hip. AB - This series of two articles is structured to provide anatomically accurate functional schematics of the motor and sensory innervation of the lower back, hip, and lower limb. This first paper provides radiographically oriented schematic axial sections of the lower back and hip in which the muscles are appropriately color-coded to match the peripheral nerves. A companion color-coded summary table allows prediction of unique patterns of denervation from 25 lesion sites. These are divided into three categories (roots T12 to S4, four plexal quadrants, and 11 sectional levels). Correlation between an imaging abnormality at one of these lesion sites and the predicted denervation pattern ensures the lesion is, in fact, clinically significant. The next article will continue this color-coded approach into the lower limb. PMID- 11473202 TI - Current status and future of metabolic cardiac imaging. PMID- 11473203 TI - Stress tests and myocardial perfusion scintigraphy. PMID- 11473204 TI - Semiquantitative analysis of SPECT images using 99Tc(m) -HMPAO in the treatment of brain perfusion after the attenuation correction by the Chang method and the application of the Butterworth filter. AB - The aim of this work was to optimize the parameters for semiquantitative analysis with regard to the average number of counts per pixel in tomographic projections. The analysed studies were divided into three groups, proportionally, to average the counts per pixel. The analysis of results proved the dependence (P<0.05) between the average geometric contrast of images with the maximum counts per pixel higher than 500, reconstructed with the cut-off frequency fc=0.50fn, and the standard image. Also, the same dependence was found between the group with an average of 200 counts per pixel and the standard image at the cut-off frequency fc=0.40fn. The analysis of the attenuation coefficient, c, showed significant differences. The attenuation coefficient c=0.12 cm-1 as accepted for further studies. Optimization of the matrix order of interpolation filter proved increasing of contrast with decreasing size of the matrix. In conclusion, according to the average number of counts per pixel in a projection the parameters of image reconstruction, particularly the cut-off frequency fc of the Butterworth filter, should be changed. Additionally, an attenuation correction with coefficient c=0.12 cm-1 and interpolation with matrix size equal 7x7 should be applied. PMID- 11473205 TI - Low frequency rTMS as an add-on antidepressive strategy: heterogeneous impact on 99mTc-HMPAO and 18 F-FDG uptake as measured simultaneously with the double isotope SPECT technique. Pilot study. AB - Single photon emission computed tomography (SPECT) and positron emission tomography (PET) are examination procedures that have shown that repetitive transcranial magnetic stimulation (rTMS) is biologically active. The aim of the present study was to investigate the patterns of regional cerebral 18F fluorodeoxyglucose (18F-FDG) uptake and regional 99mTc-hexamethylpropyleneamine oxime (99mTc-HMPAO) uptake simultaneously during a series of therapeutic rTMS at low frequency. Four drug-resistant depressed patients underwent 10 rTMS as an add on measure over 14 days. One day before and one day after TMS, simultaneous measurements of 18F-FDG, representing regional cerebral metabolic rate (rCMR), and 99mTc-HMPAO, representing regional cerebral blood flow (rCBF), were carried out. A conventional double head SPECT camera with 511 keV collimators was used. Statistically significant simultaneous overall changes of rCBF and rCMR were found in the upper prefrontal regions bilaterally in terms of increased uptake rates and in the left gyrus frontalis inferior in terms of decreased uptake rates of both isotopes compared to controls. Although this method improves our understanding of rTMS mechanism, there are limitations due to the lower resolution provided. Therapeutic rTMS seems to influence distinct, cortical regions affecting rCBF and rCMR. PMID- 11473206 TI - Whole body PET for the evaluation of bony metastases in patients with breast cancer: comparison with 99Tcm-MDP bone scintigraphy. AB - The purpose of this study was to determine the potential role of positron emission tomography (PET) using 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) for the evaluation of bony metastasis compared with 99Tcm-methylene diphosphonate (99Tcm MDP) bone scintigraphy in patients with breast cancer. Fifty-one female patients with breast cancer who had PET together with a bone scan within 1 month between September 1994 and March 1997 were included in this study. The median age was 49 years (range 29-79 years). The sensitivity, specificity and accuracy of the bone scan were 77.7%, 80.9% and 80.3%, respectively. On the other hand, for the detection of bone metastases PET had a sensitivity, specificity and accuracy of 77.7%, 97.6% and 94.1%, respectively. In the diagnosis of bony metastasis derived from breast cancer, FDG-PET was statistically superior to bone scintigraphy in its specificity. In conclusion, FDG-PET appears to be a powerful tool not only in the diagnosis of the primary lesion and soft tissue metastasis, but also in the diagnosis of bony metastasis among patients with breast cancer. PMID- 11473207 TI - Clinical features of differentiated thyroid carcinoma in children and adolescents from a sub-Himalayan iodine-deficient endemic zone. AB - Northern India is an identified sub-Himalayan iodine-deficient endemic zone. We retrospectively analysed the case files of children with differentiated thyroid carcinoma from this endemic zone and attempted to define the disease in terms of its presentation, clinical course and outcome of radioiodine therapy. Between 1967 and June 2000, 1135 patients with thyroid cancer were treated in our centre and 80 (7%) were less than 20 years of age. There were 45 females and 35 males. Histologically, 85% of patients had papillary carcinoma and the rest follicular carcinoma. Cervical lymph node involvement was seen in 66% of patients, and distant metastasis, mainly pulmonary, in 29%. In children less than 10 years of age, 75% of patients had distant metastasis at the time of presentation. The post surgery 48 h radioiodine neck uptake was 12.2+/-9.6%. Ninety-six per cent of the residual thyroid, 90% of nodal metastases and 57% of pulmonary metastases were ablated. Although nine patients had nodal recurrence between surgery and radioiodine treatment, no recurrence was observed thereafter, and three disease related deaths (all in children less than 10 years of age) were seen in the mean follow-up of 6 years. We conclude that, except for the relatively higher incidence of follicular thyroid cancer and the higher mortality in the less than 10 year age group, the course and outcome of differentiated thyroid carcinoma in children from iodine-deficient areas is no different from that in children in iodine-sufficient areas. PMID- 11473208 TI - Long-term effects of 'ecstasy' abuse on the human brain studied by FDG PET. AB - The popular recreational drug, 'ecstasy', mainly contains 3,4 methylenedioxymethamphetamine (MDMA) as the psychotropic agent. MDMA is suspected of causing neurotoxic lesions to the serotonergic system as demonstrated by animal studies, examinations of human cerebrospinal fluid, and the first positron emission tomography (PET) studies using the serotonin transporter ligand [11C] McN5652. Damage of serotonergic afferents might mediate long-lasting alterations of cerebral glucose metabolism as a secondary effect. To study a relationship between ecstasy use and long-lasting alterations, PET using 2-[18F]-fluoro-2 deoxy-d-glucose (FDG) was performed in 93 ecstasy users and 27 subjects without any known history of illicit-drug abuse. As an index of glucose metabolism, mean normalized FDG uptake was determined in both groups using a computerized brain atlas, and was compared for a selected number of brain regions. FDG uptake was normalized in each individual by dividing local FDG uptake by the maximum FDG uptake in the individual's brain. Within the group of ecstasy users we examined the relationship between FDG uptake and cumulative ecstasy dose, time since last ecstasy ingestion at the time of PET scanning, and age at first ecstasy use, respectively. Normalized FDG uptake was reduced within the striatum and amygdala of ecstasy users when compared to controls. No statistically significant correlation of the FDG uptake and the cumulative dose of ecstasy was detected. A positive correlation was found in the cingulate between FDG uptake and the time since last ecstasy ingestion. As compared to the control group, normalized FDG uptake in the cingulate was reduced in ecstasy users who took ecstasy during the last 6 months, while it was elevated in former ecstasy users who did not consume ecstasy for more than 1 year. FDG uptake was significantly more affected in ecstasy users who started to consume ecstasy before the age of 18 years. In conclusion, ecstasy abuse causes long-lasting effects on glucose metabolism in the human brain. These effects are more severe in the case of very early abuse. However, several questions still remain to be answered, i.e. the correlation of the neuronal alterations and the history of ecstasy use (cumulative dose, and time since the last dose) and its reversibility. PMID- 11473209 TI - The radiographer and role expansion in nuclear medicine. AB - With the introduction and wide application of Clinical Governance to all clinical areas, all healthcare professionals including radiographers/technologists working in Nuclear Medicine need to gain skills required for implementation. New professional and legal obligations demand that relevant expertise is developed. This article examines the potential for role expansion of radiographers/technologists in Nuclear Medicine with emphasis on the training and development aspects. The key areas addressed include clinical audit, risk management, research, management and continuing professional development. PMID- 11473210 TI - Fitting linear compartmental models by a matrix diagonalization method. AB - A general method is presented for fitting experimental data to arbitrary linear compartmental models, based on readily available public domain software. The model is defined by input data so that the same program can be used for different compartmental models. Its use is exemplified by application to the four compartment problem recently treated by Charkes and Siegel. PMID- 11473211 TI - Quality assurance of radiopharmaceuticals. Report of a joint working party: the UK Radiopharmacy Group and the NHS Pharmaceutical Quality Control Committee. AB - In this issue, the UK Radiopharmacy Group presents its third report on the Quality Assurance of Radiopharmceuticals. This concentrates on facilities which prepare only short-lived materials (shelf-life <24 hours) on site, which covers the majority of units within the UK, but is applicable to all radiopharmacies. As has come to be expected, the advice is comprehensive and provides sound interpretation of both current and potential legislation/regulations/guidelines. It will enable existing units to carry out a rapid audit of their documentation and prove invaluable to those planning new facilities. Given the litigious milieu in which we now practise, this is a timely document. PMID- 11473212 TI - Letter to the Editor. PMID- 11473213 TI - Genital ulceration in rural Uganda: sexual activity, treatment-seeking behavior, and the implications for HIV control. AB - BACKGROUND: Genital ulcer disease (GUD) is documented as a significant risk factor for HIV acquisition. Its prompt recognition and effective treatment are important components of HIV control in sub-Saharan Africa. GOALS: To determine the rate and risk factors for GUD, and to describe sexual and treatment-seeking behavior. STUDY DESIGN: A longitudinal follow-up study of 525 individuals, both those infected with HIV and those uninfected, was conducted in a population based, clinical cohort over 9 years. RESULTS: The rate of GUD in this population was 16/100 person years, and 30% of the participants reported an episode during the follow-up period. The rates were almost four times higher in participants infected with HIV than in those who were HIV-negative. Half of the participants with a regular sexual partner reported engaging in sex while symptomatic, and only 16% informed their partner. Treatment was sought for only 12% of the genital ulcer disease episodes, despite open access to doctors at a free clinic. A self diagnosis of genital herpes did not affect treatment-seeking or sexual behavior. CONCLUSIONS: Genital ulcer disease is very common in this rural population, especially among those infected with HIV. Few seek treatment, and many continue sexual activity despite symptoms without informing their partners. The implications of these findings for HIV control are a cause for concern. PMID- 11473214 TI - Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae among at-risk women, young sex workers, and street youth attending community organizations in Quebec City, Canada. AB - BACKGROUND: Despite a relatively recent decline in the global incidence of Chlamydia trachomatis and Neisseria gonorrhoeae it seems that some segments of the population such as street youth, sex workers, and individuals with social problems or delinquent behavior could be part of a core group for STDs. These persons may be reluctant to undergo STD diagnosis in traditional medical settings. GOALS: To determine the prevalence of C trachomatis and N gonorrhoeae infection using polymerase chain reaction on urine samples among subjects attending an anonymous HIV testing clinic and four community organizations in Quebec City, and to identify associated risk factors. STUDY DESIGN: A cross sectional study of 626 street youth, sex workers, and women with social problems or delinquent behavior was conducted. RESULTS: The prevalences of N gonorrhoeae and C trachomatis were, respectively, 1.1% (95% CI, 0.5%--2.3%) and 5.8% (95% CI, 4.1%--7.9%). No significant difference was found between men and women, but the sexually transmitted disease (STD) prevalence was much higher in subjects younger than 20 years: 11.4% versus 3.6% (P < 0.01). In a logistic regression model, factors independently associated with STD infection were age younger than 20 years (OR, 2.6; P = 0.007), occasional sex partners (OR, 2.9; P = 0.007), and injection of drugs (OR, 2.8; P = 0.002) in the preceding 6 months. CONCLUSIONS: A moderate STD prevalence was found in the study population. The prevalence, however, can be considered high (>10%) among street youth and young sex workers. Providing community-based STD screening and treatment services appear to be an efficient method for reaching these high-risk groups. PMID- 11473215 TI - Condom carrying is not associated with condom use and lower prevalence of sexually transmitted diseases among minority adolescent females. AB - BACKGROUND: Most of the studies associated with condom carrying and use have been conducted with adults. Because minority teenage females are particularly at risk for STD/HIV infection, further investigations specifically focusing on this population are warranted. GOAL: To determine whether observed condom carrying among adolescent females was associated with multiple measures of self-reported condom use, self-reported history of sexually transmitted diseases, and prevalence of biologically confirmed sexually transmitted diseases. METHODS: For this study, 522 sexually active African American adolescent females were recruited from low-income neighborhoods in Birmingham, Alabama. Measures of self reported condom use, STD history, and condom carrying were collected. Adolescents were also tested for three prevalent sexually transmitted diseases. RESULTS: At the time of the assessment, 8% of the adolescents were observed to have a condom with them. Condom carrying was not found to be significantly associated with condom use and prevalence of sexually transmitted diseases. CONCLUSION: Condom carrying may not be an important outcome of sexually transmitted disease/HIV prevention programs designed to reduce HIV/sexually transmitted disease risk among adolescent females. PMID- 11473216 TI - Modification of syphilitic genital ulcer manifestations by coexistent HIV infection. AB - BACKGROUND: Conflicting data exist regarding whether HIV infection leads to changes in the clinical manifestations and severity of genital ulcer disease (GUD). GOAL: To determine the impact of HIV on the etiology and clinical severity of GUD. STUDY DESIGN: From July 1990 to July 1992, consecutive patients presenting to the two Baltimore City Health Department (BCHD) Sexually Transmitted Diseases clinics were approached as candidates for enrollment in a prospective study to determine factors associated with the transmission and acquisition of sexually transmitted diseases (STDs). RESULTS: Of the 1368 patients who presented to the BCHD, 214 (16%) had genital ulcerations: 160 (21%) of 757 men and 54 (9%) of 611 women. Among the patients with GUD who had undergone HIV testing, 28 (14%) of 204 were infected with HIV: 25 (17%) of 151 men and 3 (6%) of 53 women. Although both groups-those infected with HIV and those not infected with HIV--presented with GUD of similar duration (10 versus 11 days; P = 0.17), multiple lesions were found more frequently in men with HIV infection than in uninfected men: 87% versus 62% (P = 0.02). Although not statistically significant, GUD in men with HIV infection more often were deep (64% versus 44%, respectively) rather than superficial (36% versus 57%, respectively; P = 0.08), and larger (505 mm(2) versus 109 mm 2; P = 0.06). Primary syphilis caused more GUD among men with HIV infection than among uninfected men: 9 (36%) of 25 versus 24 (19%) of 126, respectively (P < 0.01). Secondary syphilis was diagnosed with concomitant GUD more frequently among men with HIV infection than among uninfected men: 3 (13%) of 25 versus 3 (2%) of 123, respectively (P < 0.01). CONCLUSIONS: In this study, patients who presented with GUD were more likely to be infected with HIV. A higher proportion of men with HIV infection had multiple lesions, and the lesions were more likely to be caused by syphilis. PMID- 11473217 TI - Sexually transmitted disease and hiv screening guidelines for men who have sex with men. PMID- 11473218 TI - Guidance for STD clinical preventive services for persons infected with HIV. PMID- 11473219 TI - Intersecting epidemics and educable moments: sexually transmitted disease risk assessment and screening in men who have sex with men. PMID- 11473220 TI - Validity of self-reported sexually transmitted diseases among African American female adolescents participating in an HIV/STD prevention intervention trial. AB - BACKGROUND: Studies assessing the validity attributed to self-reported measures of sexually transmitted diseases (STDs) clearly are needed, particularly those used for high-risk populations such as female adolescents, in whom STD prevention is a priority. GOAL: To determine the accuracy of self-reported STD test results in female adolescents over a relatively brief period ( approximately 28 days). STUDY DESIGN: A prospective, randomized, controlled clinical trial of STD/HIV prevention for African American females, ages 14 to 18, was conducted. Study participants were recruited from medical clinics and school health classes in low income neighborhoods of Birmingham, Alabama, that had high rates of unemployment, substance abuse, violence, STDs, and teenage pregnancy. RESULTS: Of the 522 adolescents enrolled in the trial, 92% (n = 479) completed baseline STD testing and follow-up surveys. At baseline, 28% had positive test results for at least one disease: 4.8% for Neisseria gonorrhoeae, 17.1% for Chlamydia trachomatis, and 12.3% for Trichomonas vaginalis. Of the adolescents with negative STD test results, 98.8% were accurate in their self-report of STD status, as compared with 68.7% of the adolescents with positive results. Underreporting varied by type of STD. Adolescents who accurately reported their positive STD status were significantly more likely to report their receipt of treatment accurately (P < 0.001). CONCLUSIONS: The substantial underreporting of STD incidence in this study suggests that reliance on self-reports of STD history may introduce misclassification bias, potentially leading to false conclusions regarding the efficacy of prevention interventions. This observation highlights the importance of using biologic indicators as outcome measures. PMID- 11473221 TI - Association of Mycoplasma genitalium persistence in the urethra with recurrence of nongonococcal urethritis. AB - BACKGROUND: Most patients with recurrent symptomatic nongonococcal urethritis receive negative test results for Chlamydia trachomatis and Ureaplasma urealyticum, and the cause of such recurrence usually is unknown. GOAL: To assess the association of Mycoplasma genitalium with recurrent nongonococcal urethritis. STUDY DESIGN: In this study, 72 men with nongonococcal urethritis were treated with levofloxacin. Before and after treatment, symptoms and signs were assessed and first-pass urine was examined for C trachomatis, M genitalium, U urealyticum, and Mycoplasma hominis by polymerase chain reaction-based assays. RESULTS: In 6 of 45 men who had no symptoms and no evidence of inflammation after treatment, nongonococcal urethritis recurred. Of these 6 men, 5 had positive test results for M genitalium before levofloxacin treatment, which remained positive afterward. After the second treatment for recurrent nongonococcal urethritis, one man was still had a positive test result for the mycoplasma and experienced a subsequent recurrence. CONCLUSIONS: This study suggests that the persistence of M genitalium in the urethra may be associated with recurrence of nongonococcal urethritis. PMID- 11473222 TI - Adolescent chlamydia testing practices and diagnosed infections in a large managed care organization. AB - GOAL: To determine chlamydia screening practices and the resulting positive test results for adolescents enrolled in a large nonprofit managed care organization. STUDY DESIGN: The electronic medical records of all 12- to 19-year-olds enrolled in a large nonprofit managed care organization serving a demographically diverse patient population from January 1998 through December 1999 were reviewed retrospectively. RESULTS: Among the 43,205 female and 44,133 male managed care organization members, ages 12 to 19 years in 1998-1999, 7575 adolescents (8.7%) (6914 females [16%] and 661 males [1.5%]) were tested for chlamydia. Among the members tested, chlamydia was diagnosed in 1109 adolescents (14.6%) (983 females [14.2] and 126 males [19.1%]); 761 (68.6%) adolescents were retested for chlamydia; and 182 (16.4%) had repeat positive test results. The median time to diagnosis of a repeat infection was 6 months. CONCLUSIONS: Chlamydia imposes a large disease burden in the private, organized healthcare sector. Managed care organizations can use operational data to enhance chlamydia prevention services by defining testing practices and local disease prevalence. PMID- 11473224 TI - Editorial: "All true life lies in encounter." PMID- 11473223 TI - Thermoreversible gel as a candidate barrier to prevent the transmission of HIV-1 and herpes simplex virus type 2. AB - BACKGROUND: Sexually transmitted diseases (STDs) caused by HIV, herpes simplex virus (HSV), and other pathogens are spreading dramatically. The need to develop active products and vehicles to reduce this epidemic is urgent. GOAL: The efficacy of a thermoreversible gel formulation as a possible barrier to prevent the transmission of pathogens causing STDs was evaluated. STUDY DESIGN: This evaluation investigated the ability of the gel formulation to prevent infection of susceptible cells by HIV-1 and HSV-2 in vitro, the diffusion of radiolabeled herpes virus and micelles of polymer through an insertion membrane, and the electron microscopic appearance of herpes virus and gel alone or mixed together. RESULTS: The gel formulation prevents infection of susceptible cells by HIV-1 and HSV-2. It acts as an effective artificial physical barrier against the herpes virus within the first 4 hours of incubation. Herpes virus is coated by the gel or entrapped within micelles of the gel, which could hinder its attachment to target cells and inhibit its infectivity. CONCLUSION: This thermoreversible gel formulation represents an attractive matrix for the incorporation of microbicides to prevent the spread of STDs. PMID- 11473225 TI - From the Art Director's Desk: Encountering the Neuroendocrinology Letters in its new appearance. PMID- 11473226 TI - Activation of the inflammatory response system: A new look at the etiopathogenesis of major depression. AB - Major depression is accompanied by various direct and indirect indicators of a moderate activation of the inflammatory response system (IRS). Increased production of proinflammatory cytokines, such as interleukin-1 (IL-1), IL-6 and interferon (IFNgamma), may play a crucial role in the immune and acute phase response in depression. Lower serum zinc and changes in the erythron are indirect indicators of IRS activation in depression. The reciprocal relationships between IRS activation and hypothalamic-pituitary-adrenal (HPA)-axis hyperactivity, alterations in HP thyroid (HPT)-axis function and the availability of tryptophan to the brain led us to hypothesize that these neuroendocrine changes in depression are indicators of IRS activation and that a combined dysregulation of the IRS, the turnover of serotonin (5-HT) and the HPA-axis is an integral component of depression. The IRS activation model of depression provides an explanation for the psycho-social (external stress) as well as organic (internal stress) etiology of major depression. Antidepressive treatments with various antidepressive agents, including SSRIs, tricyclic and heterocyclic antidepressants, have in vivo and in vitro negative immunoregulatory effects, suggesting that their antidepressant efficacy may be attributed, in part, to their immune effects. PMID- 11473227 TI - Effects of aproteic diet on hypothalamic-pituitary- gonadal regulation in the male rat. AB - The effect of an aproteic diet (Ap) on the reproductive axis in young male rats was studied. Also the refeeding effect at different times after the aproteic diet was studied. The Ap diet was given during 21 days. In refeeding groups, the control diet was given during 2, 4 and 6 weeks after the aproteic diet. We studied the plasmatic testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels. Also the hypothalamic GnRH concentration and in vitro hypothalamic GnRH secretion in basal and induced condition was studied. The total protein deficit produced significant reduction in body, testis, seminal vesicles and prostate weights. This was accompanied with decreased levels of plasmatic testosterone (P<0.02). In this aproteic group there was a significant reduction in LH (P<0.05) and FSH (P<0.05) plasmatic levels. Refeeding with control diet reversed this situation, producing significant increment in LH (P<0.05) and FSH levels (P<0.01) at the fourth and second weeks, respectively. The basal hypothalamic GnRH secretion did not differ from the control; nevertheless the induced secretion was significantly (P<0.05) greater in the aproteic group. Also the hypothalamic GnRH concentration was increased (P<0.05) in animals fed with the aproteic diet. The minor testis, prostate, and seminal vesicles" weight, and a decreased plasmatic testosterone in rats fed with an aproteic diet, are produced by a decrease in gonadotrophin secretion. This decrease in turn is caused by a reduction in GnRH secretion, since hypothalamic GnRH concentration is increased in rats fed with the aproteic group, and induced secretion is greater in this group. All these alterations produced by an aproteic diet are reversible, since-with contol diet refeeding-the gonadotrophin secretion returned at control levels. PMID- 11473228 TI - General and unspecific damping by malignancy of the circadian amplitude of circulating human melatonin? AB - The circadian rhythm of serum melatonin of 39 cancer patients is compared with that of 28 healthy subjects matched by gender and age. Each subject provided 6 blood samples at 4-hour intervals for determination of melatonin by RIA. After log10-transformation, data series were analyzed by single and population-mean cosinor and compared between the two groups and among patients subgrouped by cancer site, stage and treatment. A circadian rhythm (P<0.001) is demonstrated for both groups, with a contributing 12-hour harmonic (P<0.001). In the absence of a difference in MESOR, the circadian amplitude of the cancer patients is smaller than that of the healthy subjects (P=0.003). Numerically, nocturnal (00:00 and 04:00) melatonin concentrations are lower and daytime (08:00-20:00) melatonin concentrations are higher in the cancer patients than in the healthy subjects (P=0.032 at 12:00 and P=0.058 at 16:00). In the age ranges examined, no differences are found with age in either group or by gender in health. No differences are found among cancer patients subgrouped either by site, stage (localized vs. metastasized) or treatment. If these results are validated, other Janus-like (two-faced: stimulation or inhibition, depending on chronome stage) effects of malignancy should be taken into consideration for screening and for timing treatment. PMID- 11473229 TI - Season's Appreciations 1998. PMID- 11473230 TI - Citation data: the wrong impact? PMID- 11473231 TI - A half-yearly aspect of circulating melatonin in pregnancies complicated by intrauterine growth retardation. AB - In investigating mechanisms underlying intrauterine growth retardation (IUGR), circulating melatonin and cortisol were radioimmunoassayed. Blood samples were collected every 4 hours during 24 hours on a strict 24-hour standardized routine in hospital from two groups of women in their third trimester of pregnancy. One group consisted of 14 healthy, uncomplicated pregnancies (HAGA); the other group consisted of 11 pregnancies complicated by intrauterine growth retardation (IUGR) confirmed at birth. The circadian characteristics of melatonin and cortisol were assessed for each woman and compared between the two groups by analyses of variance for repeated measures and by parameter tests based on the cosinor. Since a circasemiannual (about half-yearly) component prominently characterizes body weight and length at birth of children with birth characteristics below usual norms, the circadian characteristics of melatonin and cortisol were also analyzed transversely (across women within each group). The 24-hour average and the 24 hour and 12-hour amplitudes of melatonin of women in the IUGR, but not in the HAGA group, were indeed found to be modulated by an about half-yearly component. This study confirms the circadian rhythmicity of melatonin in healthy pregnant women and extends the finding to pregnancies complicated by IUGR, uncovering about half-yearly changes in melatonin in women with IUGR, thereby extending results obtained in healthy non-pregnant women and men. These variations may reflect influences from geomagnetic disturbances also characterized by a prominent half-yearly pattern, to which the pineal has been shown to be sensitive. PMID- 11473232 TI - Maternal melatonin influences rates of somatic and reproductive organs postnatal development of male rat offspring. AB - Female rat dams, housed in 12L:12D photoperiod, were pinealectomized or injected daily 1(1/2) h before onset of darkness with 250 mg melatonin/100 g BW., during pregnancy; control and pinealectomized dams received a placebo. Somatic, reproductive organs and gonadotropins levels luteinizing hormone (LH) and follicle stimulating hormone (FSH) of male offspring were examined at the following phases of their sexual development: neonate, infantile, juvenile or prepubertal and pubertal periods. Pinealectomy of the mother produced an altered developmental pattern in the offspring (PIN-X offspring). During the infantile period when pups are lacking maternal melatonin and their own melatonin rhythm is not yet established, a delayed growth of body and testis weights was observed. After the second week of life, from 15 to 25 days of age, coinciding with the initiation of the melatonin rhythm, a speed-up growth of body and testes was observed, followed by a delayed growth from 25 to 30 days, in the juvenile period; this also coinciding with reduced LH levels observed at 30 days of age. Indeed, in PIN-X offspring significantly greater growth rate was observed during the pubertal period than in control offspring, which could be due to the increase in LH secretion up to normal values observed in the PIN-X offspring. Seminal vesicles of the PIN-X offspring also showed delayed growth, which was overcome at the pubertal period. Melatonin (MEL) treatment during pregnancy produced minor alterations in postnatal development of the reproductive tract. Only increased pituitary gland weight was observed at 15 and decreased at 25 days of age. At 25 days of age, MEL offspring reached the highest LH values, and at 30 days of age, PIN-X offspring still show low values. Which suggests that other factors than the endocrine activity of the gland are affecting the somatic growth of the pituitary gland. Seminal vesicles weight was delayed at 25 days of age in the MEL offspring. These results indicate that maternal melatonin is necessary for a normal somatic growth and postnatal development of reproductive organs of the offspring. PMID- 11473233 TI - Effects of oxytocin and vasopressin on retrieval of passive avoidance response in melatonin-treated and/or pinealectomized male rats. AB - The role of the pineal gland and its hormone-melatonin-as to the impact of vasopressin (VP) and/or oxytocin (OT) on the regulation of behavior was studied, the passive avoidance task being chosen as an experimental model. The results showed that VP facilitated the avoidance latency during the first retention trial; after pinealectomy, however, VP was ineffective in this regard. Intraperitoneal application of OT was ineffective in modifying the passive avoidance latency when compared with respective saline-treated animals. Melatonin alone, when injected to shamoperated animals 30 min before behavioral experiment, did not affect the passive avoidance response in SA- or OT-treated rats, but blocked the VP-induced lengthening of the passive avoidance latency in the first retention trial. In pinealectomized and OT-treated animals the passive avoidance latency during the second retention trial was severely diminished by melatonin when compared to respective control. We conclude that: a) VP needs a regulated pineal function for developing short-term effects on the passive avoidance response and b) the effect of OT on the avoidance latency in pinealectomized rats develops after melatonin treatment as a long-term effect. PMID- 11473234 TI - Philosophy Behind Science: The European discoveries in prenatal ethnology and archeology of the mind. AB - "On ne connait que les choses que l"on apprivoise, tu deviens responsable pour toujours de ce que tu as apprivoise," Fedor-Freybergh once quoted in an editorial Saint-Exupery. I remember Gupta quoting an old Indian physician on responsibility of the parents: "The somatic and mental characteristics of the child are predetermined. Valor, health, constitution, and intellect develop in the intrauterine child due to the physiological and spiritual harmony of his parents." The responsibility is not finished by medical observations or help to make the child become intelligent. I try to bring together some of the many ideas on prenatal life which grew during decenniums in Europe to blossom in sudden or quiet revelations and which are due to functions of the new being and the threefold or manyfold way behind him and his parents. PMID- 11473235 TI - The Story Behind: Pineal mythology and chronorisk. The Swan Song of Brunetto TARQUINI. AB - Brunetto Tarquini became professor of medical semeiotics and cardiology in 1981, and chief of an internal medicine department in 1990, a position he held until his untimely death. As director of the Inter-University Center for Clinical Chronobiology and as coordinator of a post-doctoral school in chronobiology, Brunetto influenced many young Italian physicians. He became the leader of a budding specialty of chronomedicine, coordinating an international group. His focus included temporal aspects of vascular diseases from womb-to-tomb as well as oncological risk factors. His research thus ranged from neonatology over neuroendocrinology to geriatrics, by studies on the pineal in particular, documenting the signature of heliogeomagnetic master switches for circulating human melatonin. PMID- 11473236 TI - The Story Behind: Chronome/chrone. PMID- 11473237 TI - The Story Behind: Chronorisk/Circadian-Circannual (Macey, 1994). PMID- 11473238 TI - Progesterone inhibits embryotoxic effect of the complement system. AB - The complement system of normal human serum (NHS) manifests a strong, dose dependent embryotoxic potential when administered to chick embryos inducing, among others, also malformations of the brain. We have demonstrated, however, that the degree of complement-induced embryotoxicity varied remarkably in the course of the menstrual cycle of fertile healthy women, although the complement serum activity (CH100) exhibited no significant fluctuation. On the other hand, the variation of embryotoxicity appeared negatively associated with progesterone levels. Following our results high progesterone levels occurring physiologically in luteal phase of the menstrual cycle suppress the embryotoxic action of the complement system. PMID- 11473239 TI - Detection of secretory immunoglobulin A (SIgA) in saliva of ventilated and non ventilated preterm neonates. AB - The very young preterm neonate has multiple immune deficiencies which may increase his or her vulnerability to infection. Secretory Immunoglobulin A (SIgA) plays an important role in the protection of epithelial surfaces exposed to the external environment; nevertheless controversy exists with regards to the ontogeny of SIgA in newborns and especially the preterm neonate. The objective was to investigate if SIgA could be detected in the saliva of very/extremely low birthweight neonates (V/ELBW). A total of 707 samples which were collected twice daily (morning and afternoon) for three consecutive days were obtained from sixty eight preterm neonates (mean gestational age 28 weeks; conceptional age ranged from 25-35 weeks). A repeated measures design was used. Total concentration of SIgA was determined from unstimulated saliva by an Enzyme Linked Immunosorbant Assay technique. Results indicated that SIgA was detectable in the early postnatal period in the saliva of both ventilated preterms who were receiving intravenous total parenteral nutrition (TPN) and non-ventilated preterms. A 3-way repeated measures Analysis of Variance (ANOVA) showed no significant effect from 'before' and 'after' samples during a period of spontaneous activity, time and day of sampling. A significant effect of mode of nutrition was found; neonates who were receiving expressed breast milk had significantly higher concentrations of SIgA than those infants receiving TPN (df=3, F=14.27, p<0.0001). These results have implications for the care of the preterm neonate in intensive care. PMID- 11473240 TI - Opioid modulation of hypothalamic catecholaminergic neurotransmission and the pre ovulatory LH surge in the rat. AB - We have investigated the inter-relationship between the opioid and catecholaminergic systems in the control of LH secretion, and the involvement of &mgr;- and kappa-opioid subtypes in this process. Conscious female rats were intraperitoneally injected with either &mgr;- (diamorphine) or kappa-opioid agonists (U-50488H) alone or with their respective antagonists (naloxone and MR2266) before the critical period on pro-estrus. Hypothalamic catecholamine and plasma LH levels were determined by HPLC-ECD and RIA, respectively. Both &mgr;- and kappa-agonists significantly decreased concentrations of noradrenaline and its metabolite (DHPG) in all the hypothalamic regions examined concomitant with inhibition of the LH surge. Dopamine levels were selectively reduced only by the &mgr;-agonist in the MPOA. The inhibitory effects of both opioid agonists were mostly reversed following their co-administration with naloxone and MR2266 (except the kappa-antagonist on LH). These results indicate that both the &mgr;- and kappa-opioid subtypes may be involved in the inhibition of the LH surge by altering the hypothalamic noradrenaline content. PMID- 11473241 TI - Evaluation of crystallization processes in the calculi of the submandibular salivary glands. AB - Four patients were subjected to sialolithotomy of the submandibular gland in the Department of Otolaryngology, Collegium Medicum, Jagiellonian University in Cracow. The resected calculi were examined under a scanning electron microscope and chemical analysis of their structure was carried out. Results of these studies were compared with clinical observations. It has been found that sialoliths are built up by inorganic salts, mainly calcium compounds. Crystallization processes proceed at variable rates and are related to duration of the disease and intensity of inflammatory reaction. PMID- 11473242 TI - The early diagnosis of lung cancer. AB - Lung cancer is the most common fatal malignancy in both men and women, both in the United States and elsewhere in the world. Today, lung cancer is most often diagnosed on the basis of symptoms of advanced disease or when chest x-rays are taken for a variety of purposes unrelated to lung cancer detection. Unfortunately, in the United States no society or governmental agency recommends screening, even for patients with high risks, such as smokers with airflow obstruction or people with occupational exposures, including asbestos. The origins of this negative attitude toward lung cancer screening are found in 3 studies sponsored by the National Cancer Institute in the mid-1970s and conducted at Johns Hopkins University School of Medicine, the Mayo Clinic, and the Memorial Sloan-Kettering Center. These studies concluded that early identification of lung cancer through chest x-rays and cytologic diagnosis of sputum did not alter disease-specific mortality. However, patients with earlier stage disease were found through screening, which resulted in a higher resectability rate and improved survival in the screening group compared with a control group of patients receiving ordinary care. Patients in the control group often received annual chest x-rays during the course of this study, which was the standard of care at the time. Thus no true nonscreening control group resulted. The patients at highest risk were not enrolled in this study. No specific amount of pack-years of smoking intensity was required. Only men were screened. The studies were inadequately powered to show an improvement in mortality rate of less than 50%. Ninety percent of lung cancer occurs in smokers. The prevalence of lung cancer is 4 to 6 times greater when smokers have airflow obstruction than with normal airflow, when all other background factors, including smoking history, occupational risk, and family history, are the same. Screening heavy smokers (ie, > or = 30 pack-years) with airflow obstruction (forced expiratory volume in one second < 70% of normal) will yield 2% or more patients with lung cancer (prevalence cases) and, over the course of 5 years, probably from 2% to 3% of patients with additional cancers, yielding an overall incidence of 5%. New technologies include low-dose helical computed tomographic scans for small peripheral adenocarcinomas that cannot yet be visualized by standard chest x-rays and cytologic diagnosis of sputum for central squamous cell lesions. These tests are complementary. A new health care initiative, the National Lung Health Education Program, recommends spirometric testing for all smokers 45 years or older, as well as for patients with symptoms of lung cancer. Screening for lung cancer in such patients will find many cancers at an early stage when they are amenable to cure. Today, we have the knowledge and the technology that could change the outcome of lung cancer. PMID- 11473243 TI - What makes the news? PMID- 11473244 TI - Defining polyubiquitin chain topology. PMID- 11473245 TI - Implications of a fusion peptide structure. PMID- 11473246 TI - Getting a grip on how DNA polymerases function. PMID- 11473247 TI - Unfolded, yes, but random? Never! PMID- 11473248 TI - Acid test for viral fusion. PMID- 11473249 TI - Picture story. Frizzled proteins pair up. PMID- 11473251 TI - Announcement of the BioSync web site. PMID- 11473252 TI - Questions about the structure of the botulinum neurotoxin B light chain in complex with a target peptide. PMID- 11473254 TI - Structure of NaeI-DNA complex reveals dual-mode DNA recognition and complete dimer rearrangement. AB - NaeI, a novel DNA endonuclease, shows topoisomerase and recombinase activities when a Lys residue is substituted for Leu 43. The NaeI-DNA structure demonstrates that each of the two domains of NaeI recognizes one molecule of DNA duplex. DNA recognition induces dramatic rearrangements: narrowing the binding site of the Topo domain 16 A to grip DNA, widening that of the Endo domain 8 A to encircle and bend DNA 45 degrees for cleavage, and completely rebuilding the homodimer interface. The NaeI-DNA structure presents the first example of novel recognition of two copies of one DNA sequence by two different amino acid sequences and two different structural motifs in one polypeptide. PMID- 11473255 TI - Crystal structure of the human ubiquitin conjugating enzyme complex, hMms2 hUbc13. AB - The ubiquitin conjugating enzyme complex Mms2-Ubc13 plays a key role in post replicative DNA repair in yeast and the NF-kappaB signal transduction pathway in humans. This complex assembles novel polyubiquitin chains onto yet uncharacterized protein targets. Here we report the crystal structure of a complex between hMms2 (Uev1) and hUbc13 at 1.85 A resolution and a structure of free hMms2 at 1.9 A resolution. These structures reveal that the hMms2 monomer undergoes a localized conformational change upon interaction with hUbc13. The nature of the interface provides a physical basis for the preference of Mms2 for Ubc13 as a partner over a variety of other structurally similar ubiquitin conjugating enzymes. The structure of the hMms2-hUbc13 complex provides the conceptual foundation for understanding the mechanism of Lys 63 multiubiquitin chain assembly and for its interactions with the RING finger proteins Rad5 and Traf6. PMID- 11473256 TI - Structure of the Bacillus subtilis D-aminopeptidase DppA reveals a novel self compartmentalizing protease. AB - Bacillus subtilis DppA is a binuclear zinc-dependent, D-specific aminopeptidase. The X-ray structure of the enzyme has been determined at 2.4 A resolution by a three-wavelength MAD experiment. The structure reveals that DppA is a new example of a 'self-compartmentalizing protease', a family of proteolytic complexes. Proteasomes are the most extensively studied representatives of this family. The DppA enzyme is composed of identical 30 kDa subunits organized in a decamer with 52 point-group symmetry. A 20 A wide channel runs through the complex, giving access to a central chamber holding the active sites. The structure shows DppA to be a prototype of a new family of metalloaminopeptidases characterized by the SXDXEG key sequence. PMID- 11473257 TI - Structural insights into the hydrolysis of cellular nitric oxide synthase inhibitors by dimethylarginine dimethylaminohydrolase. AB - Nitric oxide synthase is inhibited by asymmetric NG-methylated derivatives of arginine whose cellular levels are controlled in part by dimethylarginine dimethylaminohydrolase (DDAH, EC 3.5.3.18). Levels of asymmetric NG,NG dimethylarginine (ADMA) are known to correlate with certain disease states. Here, the first structure of a DDAH shows an unexpected similarity to arginine:glycine amidinotransferase (EC 2.1.4.1) and arginine deiminase (EC 3.5.3.6), thus defining a superfamily of arginine-modifying enzymes. The identification of a Cys His-Glu catalytic triad and the structures of a Cys to Ser point mutant bound to both substrate and product suggest a reaction mechanism. Comparison of the ADMA DDAH and arginine-amidinotransferase complexes reveals a dramatic rotation of the substrate that effectively maintains the orientation of the scissile bond of the substrate with respect to the catalytic residues. The DDAH structure will form a basis for the rational design of selective inhibitors, which are of potential use in modulating NO synthase activity in pathological settings. PMID- 11473258 TI - Structure of beta-lactam synthetase reveals how to synthesize antibiotics instead of asparagine. AB - The enzyme beta-lactam synthetase (beta-LS) catalyzes the formation of the beta lactam ring in clavulanic acid, a clinically important beta-lactamase inhibitor. Whereas the penicillin beta-lactam ring is generated by isopenicillin N synthase (IPNS) in the presence of ferrous ion and dioxygen, beta-LS uses ATP and Mg2+ as cofactors. According to sequence alignments, beta-LS is homologous to class B asparagine synthetases (AS-Bs), ATP/Mg2+-dependent enzymes that convert aspartic acid to asparagine. Here we report the first crystal structure of a beta-LS. The 1.95 A resolution structure of Streptomyces clavuligerus beta-LS provides a fully resolved view of the active site in which substrate, closely related ATP analog alpha,beta-methyleneadenosine 5'-triphosphate (AMP-CPP) and a single Mg2+ ion are present. A high degree of substrate preorganization is observed. Comparison to Escherichia coli AS-B reveals the evolutionary changes that have taken place in beta-LS that impede interdomain reaction, which is essential in AS-B, and that accommodate beta-lactam formation. The structural data provide the opportunity to alter the synthetic potential of beta-LS, perhaps leading to the creation of new beta-lactamase inhibitors and beta-lactam antibiotics. PMID- 11473259 TI - X-ray snapshots of serine protease catalysis reveal a tetrahedral intermediate. AB - Studies on the catalytic mechanism and inhibition of serine proteases are widely used as paradigms for teaching enzyme catalysis. Ground-breaking work on the structures of chymotrypsin and subtilisin led to the idea of a conserved catalytic triad formed by the active site Ser, His and Asp residues. An oxyanion hole, consisting of the peptide amide of the active site serine and a neighbouring glycine, was identified, and hydrogen bonding in the oxyanion hole was suggested to stabilize the two proposed tetrahedral intermediates on the catalytic pathway. Here we show electron density changes consistent with the formation of a tetrahedral intermediate during the hydrolysis of an acyl-enzyme complex formed between a natural heptapeptide and elastase. No electron density for an enzyme-product complex was observed. The structures also suggest a mechanism for the synchronization of hydrolysis and peptide release triggered by the conversion of the sp2 hybridized carbonyl carbon to an sp3 carbon in the tetrahedral intermediate. This affects the location of the peptide in the active site cleft, triggering the collapse of a hydrogen bonding network between the peptide and the beta-sheet of the active site. PMID- 11473260 TI - A histone fold TAF octamer within the yeast TFIID transcriptional coactivator. AB - Gene activity in a eukaryotic cell is regulated by accessory factors to RNA polymerase II, which include the general transcription factor complex TFIID, composed of TBP and TBP-associated factors (TAFs). Three TAFs that contain histone fold motifs (yTAF17, yTAF60 and yTAF61) are critical for transcriptional regulation in the yeast Saccharomyces cerevisiae and are found in both TFIID and SAGA, a multicomponent histone acetyltransferase transcriptional coactivator. Although these three TAFs were proposed to assemble into a pseudooctamer complex, we find instead that yTAF17, yTAF60 and yTAF61 form a specific TAF octamer complex with a fourth TAF found in TFIID, yTAF48. We have reconstituted this complex in vitro and established that it is an octamer containing two copies each of the four components. Point mutations within the histone folds disrupt the octamer in vitro, and temperature-sensitive mutations in the histone folds can be specifically suppressed by overexpressing the other TAF octamer components in vivo. Our results indicate that the TAF octamer is similar both in stoichiometry and histone fold interactions to the histone octamer component of chromatin. PMID- 11473261 TI - Structure of TCTP reveals unexpected relationship with guanine nucleotide-free chaperones. AB - The translationally controlled tumor-associated proteins (TCTPs) are a highly conserved and abundantly expressed family of eukaryotic proteins that are implicated in both cell growth and the human acute allergic response but whose intracellular biochemical function has remained elusive. We report here the solution structure of the TCTP from Schizosaccharomyces pombe, which, on the basis of sequence homology, defines the fold of the entire family. We show that TCTPs form a structural superfamily with the Mss4/Dss4 family of proteins, which bind to the GDP/GTP free form of Rab proteins (members of the Ras superfamily) and have been termed guanine nucleotide-free chaperones (GFCs). Mss4 also acts as a relatively inefficient guanine nucleotide exchange factor (GEF). We further show that the Rab protein binding site on Mss4 coincides with the region of highest sequence conservation in the TCTP family. This is the first link to any other family of proteins that has been established for the TCTP family and suggests the presence of a GFC/GEF at extremely high abundance in eukaryotic cells. PMID- 11473262 TI - The laminin-binding domain of agrin is structurally related to N-TIMP-1. AB - Agrin is the key organizer of postsynaptic differentiation at the neuromuscular junction. This organization activity requires the binding of agrin to the synaptic basal lamina. Binding is conferred by the N-terminal agrin (NtA) domain, which mediates a high-affinity interaction with the coiled coil domain of laminins. Here, we report the crystal structure of chicken NtA at 1.6 A resolution. The structure reveals that NtA harbors an oligosaccharide/oligonucleotide-binding fold with several possible sites for the interaction with different ligands. A high structural similarity of NtA with the protease inhibition domain in tissue inhibitor of metalloproteinases-1 (TIMP-1) supports the idea of additional functions of agrin besides synaptogenic activity. PMID- 11473263 TI - The crystal structure of MarR, a regulator of multiple antibiotic resistance, at 2.3 A resolution. AB - MarR is a regulator of multiple antibiotic resistance in Escherichia coli. It is the prototypical member of the MarR family of regulatory proteins found in bacteria and archaea that play important roles in the development of antibiotic resistance, a global health problem. Here we describe the crystal structure of the MarR protein, determined at a resolution of 2.3 A. This is the first reported crystal structure of a member of this newly-described protein family. The structure shows MarR as a dimer with each subunit containing a winged-helix DNA binding motif. PMID- 11473264 TI - Membrane structure and fusion-triggering conformational change of the fusion domain from influenza hemagglutinin. AB - The N-terminal domain of the influenza hemagglutinin (HA) is the only portion of the molecule that inserts deeply into membranes of infected cells to mediate the viral and the host cell membrane fusion. This domain constitutes an autonomous folding unit in the membrane, causes hemolysis of red blood cells and catalyzes lipid exchange between juxtaposed membranes in a pH-dependent manner. Combining NMR structures determined at pHs 7.4 and 5 with EPR distance constraints, we have deduced the structures of the N-terminal domain of HA in the lipid bilayer. At both pHs, the domain is a kinked, predominantly helical amphipathic structure. At the fusogenic pH 5, however, the domain has a sharper bend, an additional 3(10) helix and a twist, resulting in the repositioning of Glu 15 and Asp 19 relative to that at the nonfusogenic pH 7.4. Rotation of these charged residues out of the membrane plane creates a hydrophobic pocket that allows a deeper insertion of the fusion domain into the core of the lipid bilayer. Such an insertion mode could perturb lipid packing and facilitate lipid mixing between juxtaposed membranes. PMID- 11473265 TI - Folding of malate dehydrogenase inside the GroEL-GroES cavity. AB - The chaperonin GroEL binds nonnative substrate protein in the hydrophobic central cavity of an open ring. ATP and GroES binding to the same ring converts this cavity into an encapsulated, hydrophilic chamber that mediates productive folding. A 'rack' mechanism of initial protein unfolding proposes that, upon GroES and ATP binding, the polypeptide is stretched between the binding sites on the twisting apical domains of GroEL before complete release into the chamber. Here, the structure of malate dehydrogenase (MDH) subunit during folding is monitored by deuterium exchange, peptic fragment production and mass spectrometry. When bound to GroEL, MDH exhibits a core of partially protected secondary structure that is only modestly deprotected upon ATP and GroES binding. Moreover, deprotection is broadly distributed throughout MDH, suggesting that it results from breaking hydrogen bonds between MDH and the cavity wall or global destabilization, as opposed to forced mechanical unfolding. PMID- 11473266 TI - Ripe opportunities for regional collaboration in east Asia. PMID- 11473267 TI - Back from the brink. PMID- 11473268 TI - Johns Hopkins researchers fume over government crackdown. PMID- 11473269 TI - Drugs approval process gets speed treatment. PMID- 11473271 TI - 'Political fix' saves Kyoto deal from collapse. PMID- 11473270 TI - Scientists fear new guidelines will stifle basic research. PMID- 11473272 TI - US rejects bioweapon inspections. PMID- 11473273 TI - Author of anti-encryption program faces jail. PMID- 11473275 TI - Physicists rally behind linear-collider plan. PMID- 11473276 TI - Sea lions massacred in Galapagos for sex organs. PMID- 11473278 TI - Building a biopolis. PMID- 11473280 TI - Framework welcome, but could do with fine-tuning. PMID- 11473279 TI - Picture perfect. PMID- 11473281 TI - Climate-change strategy needs to be robust. PMID- 11473282 TI - When three's not a crowd. PMID- 11473290 TI - Celebrating science. PMID- 11473291 TI - Ultradivided matter. PMID- 11473292 TI - Caught in the crossflow. PMID- 11473293 TI - Chemistry. How green was my ester. PMID- 11473294 TI - Neurobiology. Feeling bumps and holes. PMID- 11473295 TI - Condensed-matter physics. In search of soft solutions. PMID- 11473296 TI - Chemistry. Rings of destruction. PMID- 11473297 TI - Dyslexia. Talk of two theories. PMID- 11473299 TI - Planetary science. Uncool Callisto. PMID- 11473300 TI - Population ecology. Birds in a buffer state. PMID- 11473302 TI - Chemistry beyond the molecule. PMID- 11473303 TI - Persistence of visual memory for scenes. PMID- 11473304 TI - Bone histology. Evolution of growth pattern in birds. AB - Living (neornithine) birds grow up rapidly and without interruption, terminating their growth within one year and, with a few secondary exceptions, starting to fly only after or near the completion of growth. Bone histology has revealed that pre-avian theropods also grew fast for most of the postnatal period, but that this growth was usually intermittent and probably extended for more than one year. We have found surprising evidence for an early postnatal slowing-down of growth in two lineages of flying basal birds, which suggests that birds may have started their evolution as precocious fliers. PMID- 11473305 TI - Seed dispersal. Directed deterrence by capsaicin in chilies. AB - The primary function of ripe, fleshy fruit is to facilitate seed dispersal by attracting consumers, yet many fruits contain unpleasant-tasting chemicals that deter consumption by vertebrates. Here we investigate this paradox in the chili (Capsicum) and find that capsaicin, the chemical responsible for the fruit's peppery heat, selectively discourages vertebrate predators without deterring more effective seed dispersers. PMID- 11473306 TI - Nanostructure. Epitaxial diamond polytypes on silicon. AB - Carbon is unique in the variety of configurations it can adopt with itself and other elements. Here we show how ion beams can be used to nanostructure various diamond polytypes, epitaxially aligning them to a silicon substrate. The ready controllability of ion beams, which are already used to manufacture submicrometre scale devices, means that our findings should enable new carbon and non-carbon materials to be nanostructured for a host of applications. PMID- 11473307 TI - Dinosaurian growth rates and bird origins. AB - Dinosaurs, like other tetrapods, grew more quickly just after hatching than later in life. However, they did not grow like most other non-avian reptiles, which grow slowly and gradually through life. Rather, microscopic analyses of the long bone tissues show that dinosaurs grew to their adult size relatively quickly, much as large birds and mammals do today. The first birds reduced their adult body size by shortening the phase of rapid growth common to their larger theropod dinosaur relatives. These changes in timing were primarily related not to physiological differences but to differences in growth strategy. PMID- 11473308 TI - The stability against freezing of an internal liquid-water ocean in Callisto. AB - The discovery of the induced magnetic field of Callisto-one of Jupiter's moons has been interpreted as evidence for a subsurface ocean, even though the presence of such an ocean is difficult to understand in the context of existing theoretical models. Tidal heating should not be significant for Callisto, and, in the absence of such heating, it is difficult to see how this internal ocean could have survived until today without freezing. Previous work indicated that an outer ice layer on the ocean would be unstable against solid-state convection, which once begun would lead to total freezing of liquid water in about 108 years. Here I show that when a methodology for more physically reasonable water ice viscosities (that is, stress-dependent non-newtonian viscosities, rather than the stress-independent newtonian viscosities considered previously) is adopted, the outer ice shell becomes stable against convection. This implies that a subsurface ocean could have survived up to the present, without the need for invoking antifreeze substances or other special conditions. PMID- 11473309 TI - Evidence for recent climate change on Mars from the identification of youthful near-surface ground ice. AB - Ground ice in the crust and soil may be one of the largest reservoirs of water on Mars. Near-surface ground ice is predicted to be stable at latitudes higher than 40 degrees (ref. 4), where a number of geomorphologic features indicative of viscous creep and hence ground ice have been observed. Mid-latitude soils have also been implicated as a water-ice reservoir, the capacity of which is predicted to vary on a 100,000-year timescale owing to orbitally driven variations in climate. It is uncertain, however, whether near-surface ground ice currently exists at these latitudes, and how it is changing with time. Here we report observational evidence for a mid-latitude reservoir of near-surface water ice occupying the pore space of soils. The thickness of the ice-occupied soil reservoir (1-10 m) and its distribution in the 30 degrees to 60 degrees latitude bands indicate a reservoir of (1.5-6.0) x 104 km3, equivalent to a global layer of water 10-40 cm thick. We infer that the reservoir was created during the last phase of high orbital obliquity less than 100,000 years ago, and is now being diminished. PMID- 11473310 TI - In situ detection of collisionless reconnection in the Earth's magnetotail. AB - Magnetic reconnection is the process by which magnetic field lines of opposite polarity reconfigure to a lower-energy state, with the release of magnetic energy to the surroundings. Reconnection at the Earth's dayside magnetopause and in the magnetotail allows the solar wind into the magnetosphere. It begins in a small 'diffusion region', where a kink in the newly reconnected lines produces jets of plasma away from the region. Although plasma jets from reconnection have previously been reported, the physical processes that underlie jet formation have remained poorly understood because of the scarcity of in situ observations of the minuscule diffusion region. Theoretically, both resistive and collisionless processes can initiate reconnection, but which process dominates in the magnetosphere is still debated. Here we report the serendipitous encounter of the Wind spacecraft with an active reconnection diffusion region, in which are detected key processes predicted by models of collisionless reconnection. The data therefore demonstrate that collisionless reconnection occurs in the magnetotail. PMID- 11473311 TI - Quantum-enhanced positioning and clock synchronization. AB - A wide variety of positioning and ranging procedures are based on repeatedly sending electromagnetic pulses through space and measuring their time of arrival. The accuracy of such procedures is classically limited by the available power and bandwidth. Quantum entanglement and squeezing have been exploited in the context of interferometry, frequency measurements, lithography and algorithms. Here we report that quantum entanglement and squeezing can also be employed to overcome the classical limits in procedures such as positioning systems, clock synchronization and ranging. Our use of frequency-entangled pulses to construct quantum versions of these protocols results in enhanced accuracy compared with their classical analogues. We describe in detail the problem of establishing a position with respect to a fixed array of reference points. PMID- 11473312 TI - Diamagnetic activity above Tc as a precursor to superconductivity in La2-xSrxCuO4 thin films. AB - Superconductors show zero resistance to electric current, and expel magnetic flux (the Meissner effect) below the transition temperature (Tc). In conventional superconductors, the 'Cooper pairs' of electrons that are responsible for superconductivity form only below Tc. In the unconventional high-Tc superconductors, however, a strong electron correlation is essential for pair formation: there is evidence that some pairs are formed above Tc in samples that have less than the optimal density of charge carriers (underdoped) and an energy gap-the 'pseudogap'-appears to be present. Moreover, excitations that look like the vortices that carry magnetic flux inside the superconducting state have been reported above Tc (refs 6, 7). Although the origin of the pseudogap remains controversial, phase fluctuations above Tc, leading to some form of local superconductivity or local pairing, seem essential. Here we report magnetic imaging (scanning SQUID microscopy) of La2-xSrxCuO4 thin films. Clear quantized vortex patterns are visible below Tc (18-19 K), and we observe inhomogeneous magnetic domains that persist up to 80 K. We interpret the data as suggesting the existence of diamagnetic regions that are precursors to the Meissner state. PMID- 11473313 TI - Sn-zeolite beta as a heterogeneous chemoselective catalyst for Baeyer-Villiger oxidations. AB - The Baeyer-Villiger oxidation, first reported more than 100 years ago, has evolved into a versatile reaction widely used to convert ketones-readily available building blocks in organic chemistry-into more complex and valuable esters and lactones. Catalytic versions of the Baeyer-Villiger oxidation are particularly attractive for practical applications, because catalytic transformations simplify processing conditions while minimizing reactant use as well as waste production. Further benefits are expected from replacing peracids, the traditionally used oxidant, by cheaper and less polluting hydrogen peroxide. Dissolved platinum complexes and solid acids, such as zeolites or sulphonated resins, efficiently activate ketone oxidation by hydrogen peroxide. But these catalysts lack sufficient selectivity for the desired product if the starting material contains functional groups other than the ketone group; they perform especially poorly in the presence of carbon-carbon double bonds. Here we show that upon incorporation of 1.6 weight per cent tin into its framework, zeolite beta acts as an efficient and stable heterogeneous catalyst for the Baeyer Villiger oxidation of saturated as well as unsaturated ketones by hydrogen peroxide, with the desired lactones forming more than 98% of the reaction products. We ascribe this high selectivity to direct activation of the ketone group, whereas other catalysts first activate hydrogen peroxide, which can then interact with the ketone group as well as other functional groups. PMID- 11473314 TI - Warm tropical ocean surface and global anoxia during the mid-Cretaceous period. AB - The middle of the Cretaceous period (about 120 to 80 Myr ago) was a time of unusually warm polar temperatures, repeated reef-drowning in the tropics and a series of oceanic anoxic events (OAEs) that promoted both the widespread deposition of organic-carbon-rich marine sediments and high biological turnover. The cause of the warm temperatures is unproven but widely attributed to high levels of atmospheric greenhouse gases such as carbon dioxide. In contrast, there is no consensus on the climatic causes and effects of the OAEs, with both high biological productivity and ocean 'stagnation' being invoked as the cause of ocean anoxia. Here we show, using stable isotope records from multiple species of well-preserved foraminifera, that the thermal structure of surface waters in the western tropical Atlantic Ocean underwent pronounced variability about 100 Myr ago, with maximum sea surface temperatures 3-5 degrees C warmer than today. This variability culminated in a collapse of upper-ocean stratification during OAE-1d (the 'Breistroffer' event), a globally significant period of organic-carbon burial that we show to have fundamental, stratigraphically valuable, geochemical similarities to the main OAEs of the Mesozoic era. Our records are consistent with greenhouse forcing being responsible for the warm temperatures, but are inconsistent both with explanations for OAEs based on ocean stagnation, and with the traditional view (reviewed in ref. 12) that past warm periods were more stable than today's climate. PMID- 11473315 TI - Dinosaurian growth patterns and rapid avian growth rates. AB - Did dinosaurs grow in a manner similar to extant reptiles, mammals or birds, or were they unique? Are rapid avian growth rates an innovation unique to birds, or were they inherited from dinosaurian precursors? We quantified growth rates for a group of dinosaurs spanning the phylogenetic and size diversity for the clade and used regression analysis to characterize the results. Here we show that dinosaurs exhibited sigmoidal growth curves similar to those of other vertebrates, but had unique growth rates with respect to body mass. All dinosaurs grew at accelerated rates relative to the primitive condition seen in extant reptiles. Small dinosaurs grew at moderately rapid rates, similar to those of marsupials, but large species attained rates comparable to those of eutherian mammals and precocial birds. Growth in giant sauropods was similar to that of whales of comparable size. Non-avian dinosaurs did not attain rates like those of altricial birds. Avian growth rates were attained in a stepwise fashion after birds diverged from theropod ancestors in the Jurassic period. PMID- 11473316 TI - Mealybug beta-proteobacterial endosymbionts contain gamma-proteobacterial symbionts. AB - Some insects have cultivated intimate relationships with mutualistic bacteria since their early evolutionary history. Most ancient 'primary' endosymbionts live within the cytoplasm of large, polyploid host cells of a specialized organ (bacteriome). Within their large, ovoid bacteriomes, mealybugs (Pseudococcidae) package the intracellular endosymbionts into 'mucus-filled' spheres, which surround the host cell nucleus and occupy most of the cytoplasm. The genesis of symbiotic spheres has not been determined, and they are structurally unlike eukaryotic cell vesicles. Recent molecular phylogenetic and fluorescent in situ hybridization (FISH) studies suggested that two unrelated bacterial species may share individual host cells, and that bacteria within spheres comprise these two species. Here we show that mealybug host cells do indeed harbour both beta- and gamma-subdivision Proteobacteria, but they are not co-inhabitants of the spheres. Rather, we show that the symbiotic spheres themselves are beta-proteobacterial cells. Thus, gamma-Proteobacteria live symbiotically inside beta-Proteobacteria. This is the first report, to our knowledge, of an intracellular symbiosis involving two species of bacteria. PMID- 11473317 TI - The buffer effect and large-scale population regulation in migratory birds. AB - Buffer effects occur when sites vary in quality and fluctuations in population size are mirrored by large changes in animal numbers in poor-quality sites but only small changes in good-quality sites. Hence, the poor sites 'buffer' the good sites, a mechanism that can potentially drive population regulation if there are demographic costs of inhabiting poor sites. Here we show that for a migratory bird this process can apply on a country-wide scale with consequences for both survival and timing of arrival on the breeding grounds (an indicator of reproductive success). The Icelandic population of the black-tailed godwit, Limosa limosa islandica, wintering in Britain has increased fourfold since the 1970s (ref. 5) but rates of change within individual estuaries have varied from zero to sixfold increases. In accordance with the buffer effect, rates of increase are greater on estuaries with low initial numbers, and godwits on these sites have lower prey-intake rates, lower survival rates and arrive later in Iceland than godwits on sites with stable populations. The buffer effect can therefore be a major process influencing large-scale population regulation of migratory species. PMID- 11473318 TI - Crossflow filtration in suspension-feeding fishes. AB - Rows of comb-like or tufted gill rakers in the oral cavity of suspension-feeding fishes (for example, herring, anchovies and tilapia) have been thought to serve as (1) non-porous barriers that direct particle-laden water to the sticky oral roof, where particles are retained as water exits from the oral cavity, (2) conventional dead-end filters that sieve particles from water exiting between rakers, or (3) sticky filters that retain particles encountered by a hydrosol filtration mechanism. Here we present data from computational fluid dynamics and video endoscopy in suspension-feeding fish indicating that the rakers of three distantly related species function instead as a crossflow filter. Particles are concentrated inside the oral cavity as filtrate exits between the rakers, but particles are not retained on the rakers. Instead, the high-velocity crossflow along the rakers carries particles away from the raker surfaces and transports the particles towards the oesophagus. This crossflow prevents particles from clogging the gaps between the rakers, and solves the mystery of particle transport from the rakers to the oesophagus. PMID- 11473319 TI - Natural conjugative plasmids induce bacterial biofilm development. AB - Horizontal gene transfer is a principal source of evolution leading to change in the ecological character of bacterial species. Bacterial conjugation, which promotes the horizontal transfer of genetic material between donor and recipient cells by physical contact, is a phenomenon of fundamental evolutionary consequence. Although conjugation has been studied primarily in liquid, most natural bacterial populations are found associated with environmental surfaces in complex multispecies communities called biofilms. Biofilms are ideally suited to the exchange of genetic material of various origins, and it has been shown that bacterial conjugation occurs within biofilms. Here I investigate the direct contribution of conjugative plasmids themselves to the capacity of the bacterial host to form a biofilm. Natural conjugative plasmids expressed factors that induced planktonic bacteria to form or enter biofilm communities, which favour the infectious transfer of the plasmid. This general connection between conjugation and biofilms suggests that medically relevant plasmid-bearing strains are more likely to form a biofilm. This may influence both the chances of biofilm related infection risks and of conjugational spread of virulence factors. PMID- 11473320 TI - Force can overcome object geometry in the perception of shape through active touch. AB - Haptic (touch) perception normally entails an active exploration of object surfaces over time. This is called active touch. When exploring the shape of an object, we experience both geometrical and force cues. For example, when sliding a finger across a surface with a rigid bump on it, the finger moves over the bump while being opposed by a force whose direction and magnitude are related to the slope of the bump. The steeper the bump, the stronger the resistance. Geometrical and force cues are correlated, but it has been commonly assumed that shape perception relies on object geometry alone. Here we show that regardless of surface geometry, subjects identified and located shape features on the basis of force cues or their correlates. Using paradoxical stimuli, for example combining the force cues of a bump with the geometry of a hole, we found that subjects perceived a bump. Conversely, when combining the force cues of a hole with the geometry of a bump, subjects typically perceived a hole. PMID- 11473321 TI - Ubiquitination-dependent mechanisms regulate synaptic growth and function. AB - The covalent attachment of ubiquitin to cellular proteins is a powerful mechanism for controlling protein activity and localization. Ubiquitination is a reversible modification promoted by ubiquitin ligases and antagonized by deubiquitinating proteases. Ubiquitin-dependent mechanisms regulate many important processes including cell-cycle progression, apoptosis and transcriptional regulation. Here we show that ubiquitin-dependent mechanisms regulate synaptic development at the Drosophila neuromuscular junction (NMJ). Neuronal overexpression of the deubiquitinating protease fat facets leads to a profound disruption of synaptic growth control; there is a large increase in the number of synaptic boutons, an elaboration of the synaptic branching pattern, and a disruption of synaptic function. Antagonizing the ubiquitination pathway in neurons by expression of the yeast deubiquitinating protease UBP2 (ref. 5) also produces synaptic overgrowth and dysfunction. Genetic interactions between fat facets and highwire, a negative regulator of synaptic growth that has structural homology to a family of ubiquitin ligases, suggest that synaptic development may be controlled by the balance between positive and negative regulators of ubiquitination. PMID- 11473322 TI - Antibacterial agents based on the cyclic D,L-alpha-peptide architecture. AB - The rapid emergence of bacterial infections that are resistant to many drugs underscores the need for new therapeutic agents. Here we report that six- and eight-residue cyclic d,l-alpha-peptides act preferentially on Gram-positive and/or Gram-negative bacterial membranes compared to mammalian cells, increase membrane permeability, collapse transmembrane ion potentials, and cause rapid cell death. The effectiveness of this class of materials as selective antibacterial agents is highlighted by the high efficacy observed against lethal methicillin-resistant Staphylococcus aureus infections in mice. Cyclic d,l-alpha peptides are proteolytically stable, easy to synthesize, and can be derived from a potentially vast membrane-active sequence space. The unique abiotic structure of the cyclic peptides and their quick bactericidal action may also contribute to limit temporal acquirement of drug resistant bacteria. The low molecular weight d,l-alpha-peptides offer an attractive complement to the current arsenal of naturally derived antibiotics, and hold considerable potential in combating a variety of existing and emerging infectious diseases. PMID- 11473323 TI - RPA governs endonuclease switching during processing of Okazaki fragments in eukaryotes. AB - Extensive work on the maturation of lagging strands during the replication of simian virus 40 DNA suggests that the initiator RNA primers of Okazaki fragments are removed by the combined action of two nucleases, RNase HI and Fen1, before the Okazaki fragments join. Despite the well established in vitro roles of these two enzymes, genetic analyses in yeast revealed that null mutants of RNase HI and/or Fen1 are not lethal, suggesting that an additional enzymatic activity may be required for the removal of RNA. One such enzyme is the Saccharomyces cerevisiae Dna2 helicase/endonuclease, which is essential for cell viability and is well suited to removing RNA primers of Okazaki fragments. In addition, Dna2 interacts genetically and physically with several proteins involved in the elongation or maturation of Okazaki fragments. Here we show that the endonucleases Dna2 and Fen1 act sequentially to facilitate the complete removal of the primer RNA. The sequential action of these enzymes is governed by a single stranded DNA-binding protein, replication protein-A (RPA). Our results demonstrate that the processing of Okazaki fragments in eukaryotes differs significantly from, and is more complicated than, that occurring in prokaryotes. We propose a novel biochemical mechanism for the maturation of eukaryotic Okazaki fragments. PMID- 11473324 TI - Prevention of pancreatitis in patients with idiopathic recurrent pancreatitis: a prospective nonblinded randomized study using endoscopic stents. AB - BACKGROUND AND STUDY AIMS: Currently there is no available therapy to prevent attacks of acute pancreatitis in patients with idiopathic recurrent pancreatitis (IRP). This randomized, nonblinded prospective, controlled trial was undertaken to evaluate the effectiveness of pancreatic duct stents in preventing attacks of pancreatitis in IRP. PATIENTS AND METHODS: During a 5-year period 34 patients met the diagnostic criteria for IRP. Patients were randomly assigned to one of two groups; 19 patients (14 women, 5 men, mean age 44) to the pancreatic stent group; and 15 patients (10 women, five men, mean age 47) to the control group. The stent group received three stents over a period of 1 year and the control group had selective pancreatograms but no stent. Mean follow-up was 33 months (range 13-77) and 35 months (range 10-78) in the stent and control groups, respectively. Episodes of pancreatitis, frequency and intensity of pain requiring emergency room visits, and hospitalizations were recorded. RESULTS: Recurrence of pancreatitis occurred in eight out of 15 patients (53%) in the control group, but in only two our of 19 patients (11%) in the stent group (P<0.02). Two patients in the control group who had recurrences of pancreatitis crossed over to stent therapy and had no further pancreatitis thereafter. Six patients each, 32% and 40% in the stent and control groups respectively, continued to have pancreatic type pain. In the study period 17 stents were occluded and 14 migrated out. CONCLUSION: The results of this study suggest that pancreatic duct stenting may prevent recurrent attacks of pancreatitis in IRP patients. Intermittent pancreatic duct sphincter dysfunction or relative outlet obstruction may be the underlying cause for the recurrent attacks of pancreatitis. PMID- 11473325 TI - Endoscopic papillary balloon dilation vs. sphincterotomy for removal of common bile duct stones: a prospective randomized pilot study. AB - BACKGROUND AND STUDY AIMS: Endoscopic sphincterotomy (EST) is the standard procedure for gaining access to the common bile duct for removal of bile duct stones. However, the procedure is associated with both short-term and long-term complications. Recent reports have described the use of endoscopic papillary balloon dilation (EBD) as an effective and safe alternative to EST. We conducted a prospective randomized pilot study to compare the efficacy and short-term complication rates of these two established methods for removing uncomplicated bile duct stones. PATIENTS AND METHODS: Sixty patients were randomly assigned to receive either EST (n = 30) or EBD (n = 30) prior to removal of bile duct stones (maximum size 20 mm, maximum number five). The patient groups were comparable with regard to sex and age ratios, the size of the stones (EST: mean 10 +/- 4.7 mm; EBD: mean 7 +/- 3.5 min; not significant) and the numbers of stones (EST: mean 1.8 +/- 1.5 mm; EBD: mean 1.6 +/- 1.1 mm; not significant). EBD was carried out using a balloon-tipped biliary catheter (Maxforce, Microvasive, Boston, Massachusetts, USA) with a maximum diameter of 24 Fr for 45-60 s. Bile duct stones were removed using Dormia baskets or retrieval balloons, or both. RESULTS: The two methods were successful in all patients studied. Subsequent stone removal was possible in all 30 patients after EST (100%) and in 23 of the 30 who underwent EBD (77%), respectively (P<0.01). After conversion to EST, complete bile duct clearance was also achieved in the remaining seven EBD patients. The mean duration for the whole procedure was 17 +/- 12 min for EST and 29 +/- 15 min for EBD (not significant). Complications (WHO grades 2-4) were observed in five of the 30 EST patients (three cases of mild pancreatitis, two of hemorrhage) and in nine of the 30 EBD patients (three cases of cholangitis, four of mild pancreatitis, and two of severe pancreatitis), showing a trend toward higher complication rates in the EBD group. Postintervention hyperamylasemia was observed in six patients (three in each group). CONCLUSIONS: The results of this prospective randomized pilot study indicate that EST is superior to EBD in terms of stone removal, duration of the procedure, and complication rates. EST will therefore continue to be the standard procedure for stone removal in the near future. Further studies will be needed in order to compare the longer-term results with EST and EBD. PMID- 11473326 TI - Lifting of lesions during endoscopic mucosal resection (EMR) of early colorectal cancer: implications for the assessment of resectability. AB - BACKGROUND AND STUDY AIMS: This study assessed the indications for and limitations of endoscopic mucosal resection (EMR) for early colorectal cancer, focusing on the way in which the lesion lifts after submucosal injection. PATIENTS AND METHODS: The study included 94 patients with early colorectal cancer who received EMR treatment. The lifting of the lesion after submucosal injection was analyzed (classified as completely lifted/soft; completely lifted/hard; incompletely lifted; and non-lifted) along with the endoscopic findings, pathological findings, and clinical course. RESULTS: Almost all completely lifted/soft lesions were mucosal cancers. Some of the completely lifted/hard lesions were staged as sm2. The incompletely lifted lesions included stages sm1 to sm3. Non-lifting lesions were almost always deeper than sm3. The lifting condition was significantly associated with the depth of invasion, and the lesion type was related to the extent of lifting but not to tumor size or recurrent disease. Recurrent disease was noted in three patients who underwent piecemeal EMR. CONCLUSIONS: The indication for EMR is easily assessed on the basis of the lifting characteristics of the tumor after submucosal injection, which was found to be significantly related to the depth of invasion. The factor limiting the indication for EMR is not the size of a tumor, but its lifting condition. PMID- 11473327 TI - Right-sided shift found in metachronous colorectal adenomas. AB - BACKGROUND AND STUDY AIMS: Patients who have had a colorectal adenoma are likely to develop a metachronous adenoma, and therefore need to be kept under surveillance. The question is whether metachronous adenomas will be found at the same anatomical site as the preceding adenomas, thus prompting us to focus surveillance examinations on this region, using flexible sigmoidoscopy or total colonoscopy. PATIENTS AND METHODS: Between 1978 and 1996 a long-term follow up of 1091 patients was prospectively documented at the Erlangen Registry of Colorectal Polyps. The anatomical sites--distal (rectosigmoid) and proximal colon--of two subsequent generations of metachronous adenomas were analysed using logistic regression analysis. RESULTS: In 556 (51%) patients metachronous adenomas were found during follow up. In 211 (37.9%) of these patients a right-sided shift in the first generation of metachronous adenomas in comparison with initial lesions (P<0.0001) was found. Some 305 patients underwent further follow up, and 51 (27.1%) out of 188 patients with metachronous lesions demonstrated a right-sided shift in their second generation (vs. first generation) of metachronous adenomas. Proximally located first generation metachronous adenomas would be missed by flexible sigmoidoscopy in 50.8% of patients with only distal adenomas at baseline colonoscopy, and in 58.7% of patients with second-generation lesions. Multivariate analysis revealed that multiplicity (odds ratio 2.64, 95% CI 1.49 4.66) and size of initial adenomas (>5 mm) (odds ratio 3.60, 95% CI 1.96-6.66) were significantly associated with a right-sided shift in metachronous adenomas, while female gender was associated with a significantly lower tendency to manifest a right-sided shift (odds ratio of 0.64. 95% CI 0.43-0.94). CONCLUSIONS: Metachronous adenomas are found significantly more often in the right colon than would be expected assuming clustering at the same anatomical site as the preceding lesions. Total colonoscopy is thus needed for surveillance, regardless of the initial adenoma site. PMID- 11473328 TI - The indications, utilization and safety of gastrointestinal endoscopy in an extremely elderly patient cohort. AB - BACKGROUND: In developed nations, increasing proportions of the population now reach advanced age. Physicians may be reluctant to refer such patients for non critical diagnostic and therapeutic interventions, on the basis of perceived diminution of tolerance, safety and substantive benefits in these patients. We aimed to review the utility and safety of gastrointestinal endoscopy in an extremely elderly cohort. METHODS: The study involved 214 consecutive participants aged 85 years or more, between 1995 and 1997. They were identified using a prospective database linked to the endoscopy reporting system. Procedure type, indication, use of sedation, complications, and outcomes were evaluated. RESULTS: The median age was 87 (85-94, sigma = 2). The female:male ratio was 3:2; 185 had undergone one procedure and 29 two or more; and 65% of procedures were performed on an outpatient basis. Of the inpatient procedures, 10% of all procedures were performed emergently, predominantly for upper gastrointestinal hemorrhage. Midazolam was administered to 129 patients (60%), at a median dose of 2 mg (range 1-11); of these, 75 (35%) also received a median dose of 25 microg fentanyl (range 12.5-125). Colonoscopy (n = 95) was the most frequently performed procedure, followed by esophagogastroduodenoscopy (EGD) (n = 64) and endoscopic retrograde cholangiopancreatography (ERCP) (n = 21). There was no procedure related mortality. The incidence of post-ERCP pancreatitis was 5%, colonic perforation 1%, and cardiopulmonary complications in sedated patients, 0.6%. The majority underwent procedures which related to active management of ongoing medical problems, and procedures were performed for palliative indications in only 15 (7%) patients. CONCLUSIONS: Gastrointestinal endoscopy is extremely safe and well tolerated in extremely elderly patients. Age alone should not influence decisions relating to its utilization. PMID- 11473329 TI - Initial experience with new dedicated needles for laparoscopic ultrasound-guided fine-needle aspiration and histological biopsies. AB - BACKGROUND AND STUDY AIMS: Laparoscopic ultrasonography (LUS) is an important imaging modality during laparoscopic staging of intra-abdominal malignancies, but LUS-assisted biopsy is often difficult or impossible. We report a newly developed inbuilt biopsy system for direct LUS-guided fine-needle aspiration (FNA) and Tru cut biopsies. PATIENTS AND METHODS: LUS-guided biopsy was performed in 20 patients with upper gastrointestinal tract tumors. The biopsied lesions had either not been previously detected by other imaging modalities or had been inaccessible, or the biopsy sample had been inadequate. Primary diagnosis, duration of biopsy procedure, needle monitoring (visibility, penetration, and deviation), complications, technical failures, and pathological findings were prospectively recorded. RESULTS: 44 biopsies were performed with 25 needles (19, 20, and 22-G). Needle monitoring and penetration were good or acceptable in 18 patients (90%). Slight needle deviation (<10 mm) was seen in eight patients (40%). The LUS-guided biopsy specimen was sufficient for analysis in 13 patients (65%). In two additional patients, adequate material was obtained, but pathological examination was impossible owing to incorrect handling of the specimen. The biopsy procedures lasted 16.3 minutes (range 10-20 minutes) and no complications were seen. CONCLUSIONS: LUS-guided fine-needle aspiration or Trucut biopsy is possible using this newly developed biopsy system. These preliminary data suggest that LUS-guided biopsy may further improve the diagnostic possibilities of LUS. PMID- 11473330 TI - Laparoscopic repair of paraesophageal hernia. AB - BACKGROUND AND STUDY AIMS: Surgical repair of paraesophageal hernia is mandatory, due to the risk of severe complications, and it can be accomplished via the laparoscopic route. This study presents the results of laparoscopic repair of paraesophageal hernia combined with anterior hemifundoplication. PATIENTS AND METHODS: During a two-year period, ten consecutive patients with paraesophageal hernia (six men, four women; mean age 73, range 55-82) underwent laparoscopic treatment. Five patients presented with symptoms of gastroesophageal reflux, while another four reported lower chest pain. There was one patient in whom the paraesophageal hernia was manifested with upper gastrointestinal bleeding. Six patients had type III hiatal hernia. They all underwent esophagography, upper gastrointestinal endoscopy, stationary manometry, and 24-hour ambulatory pH metry, preoperatively and within three months postoperatively. At laparoscopy, the hernia content was completely reduced, the sac excised, and the diaphragmatic crura approximated. The operation was completed with an anterior hemifundoplication. In three cases, a prosthetic mesh was applied to close the hiatal defect securely. RESULTS: Operating times ranged from 75 min to 125 min (mean 90 min). There were no postoperative deaths. One patient developed atelectasis, and another had empyema of the left pleura, treated with drainage and antibiotics. All patients but one were discharged on the second or third postoperative day. At the three-month follow-up examination, none of the patients had symptoms related to the paraesophageal hernia, gastroesophageal reflux, or fundoplication. Esophagography demonstrated restoration of normal anatomy at the gastroesophageal region, while esophageal motility was improved, and esophageal pH-metry showed no gastroesophageal reflux. CONCLUSION: Laparoscopic repair of paraesophageal hernias is a safe, technically feasible, and well tolerated procedure, which offers rapid and total relief of symptoms. The addition of an anterior hemifundoplication not only cures preexisting gastroesophageal reflux, but also prevents the development of postoperative gastroesophageal reflux. PMID- 11473331 TI - Changes in esophageal variceal pressure after transcatheter arterial embolization for hepatocellular carcinoma. AB - BACKGROUND AND STUDY AIM: The aim of this study was to investigate the influence of transcatheter arterial embolization on esophageal variceal pressure and portal hemodynamics. PATIENTS AND METHODS: Out of 18 cirrhotic patients with hepatocellular carcinoma, 12 underwent transcatheter arterial embolization and the remaining six patients underwent angiography alone as a control. We examined esophageal variceal pressure with an endoscopic pneumatic pressure sensor and portal blood flow with Doppler ultrasonography immediately before and 3 days after transcatheter arterial embolization or angiography. RESULTS: Angiography alone did not influence esophageal variceal pressure or portal blood flow. Transcatheter arterial embolization resulted in an increase in variceal pressure in five (42%) of the 12 patients and in a marked increase in portal blood flow in eight (88.9%) of nine patients, although no change in the endoscopic variceal findings was observed after transcatheter arterial embolization. The change in esophageal variceal pressure did not correlate with the change in portal blood flow. We could not find predictive factors for the elevation of variceal pressure after transcatheter arterial embolization. CONCLUSION: Our study demonstrated that transcatheter arterial embolization resulted in an increase in esophageal variceal pressure in about half of the patients, bearing little relation to portal hemodynamic parameters. PMID- 11473332 TI - Self-expanding oesophageal stents: strategies for re-intervention. AB - BACKGROUND AND STUDY AIMS: Self-expanding metal stents have become accepted palliation for inoperable malignant oesophageal obstruction, the cost of the devices being offset against the ease of insertion and the reduced complication rate. However, re-intervention is often required for obstruction, malposition, migration and tumour progression. The marginal cost of re-stenting is generally higher than other modalities. This study aims to determine the rate of re intervention and the effectiveness of the various intervention modalities. PATIENTS AND METHODS: A population of 165 patients, treated in a tertiary referral oesophageal centre, (132 with oesophageal cancer, 31 with mediastinal metastases from other tumours, two with benign conditions) whose initial stent placement was performed between January 1994 and December 1998 was followed-up through July 1999 or till death. RESULTS: A total of 75 re-interventions were required in 44 patients and were successful in 51 (68%). Rigid oesophagoscopy and removal of food bolus was successful in three out of three, dilation in one of 11, rigid oesophagoscopy and physical debridement in 12 of 17 and laser debridement in 12 of 20. Re-stenting was the primary re-intervention in 10 cases and was ultimately necessary in 14 patients (with 11 self-expanding metal stents, three Celestin) who had previously undergone other forms of re-intervention. It was not successful in one case. The median survival following first re intervention was 9.8 weeks (compared with 14.3 weeks for initial stenting) and was longer in those receiving radiotherapy (23.6 weeks) or chemotherapy (14.4 weeks). CONCLUSIONS: While repeated stenting is usually successful, debridement and laser vaporization are viable alternatives for proximal tumour overgrowth or ingrowth in the upper or middle third of the oesophagus. Distal tumour growth or ingrowth at the oesophagogastric junction are best treated with a second stent. Repeated treatment is justified, as survival following first re-intervention is comparable to that after initial stenting, particularly in those patients who are able to undergo chemotherapy or radiotherapy. PMID- 11473333 TI - Colonoscopic manifestations of primary colorectal lymphoma. AB - BACKGROUND AND STUDY AIMS: Little is known about the colonoscopic manifestation of colorectal lymphoma. This article describes the use of colonoscopy for diagnosis of primary colorectal lymphoma. PATIENTS AND METHODS: We analyzed colonoscopic findings in 13 retrospectively collected cases of primary colorectal lymphoma diagnosed between 1990 and 1999 at our hospital. RESULTS: The primary colorectal tymphomas were distinguished into three colonoscopic types: a) the mucosal type, which included six lymphomas (46%), which were classified into two subtypes, erosive (two lymphomas) and ulcerative (four tymphomas); b) the polypoid type, which included three tymphomas (23%); and c) the massive type, which included four (31%). The proportion of patients with T-cell lineage lymphoma (5 of 12, 42%) was higher than in previous studies. Three cases were diagnosed as enteropathy-associated T-cell (EAT)-like lymphoma: one presenting with multicentric ulcerations, one as a massive type and another as a polypoid type. Of those with B-cell lineage lymphoma (7 of 12, 58%), three presented as massive type, two as polypoid type and two as erosive subtype. CONCLUSIONS: Based on the analysis of colonoscopic records, we have proposed three main types of primary colorectal lymphoma. An unusually high proportion of T-cell lineage lymphomas was noted, but as the number of cases was low, further study is still needed for confirmation. PMID- 11473334 TI - The laparoscopic Janeway gastrostomy. An alternative technique when percutaneous endoscopic gastrostomy is impractical. AB - BACKGROUND AND STUDY AIMS: Today, percutaneous endoscopic gastrostomy (PEG) is the standard procedure for ensuring safe feeding access. In the case of advanced hypopharyngeal and esophageal carcinomas, it may not be feasible to pass an existing stenosis by endoscopy. The aim of this prospective study was to investigate the laparoscopic Janeway gastrostomy as an alternative technique for guaranteeing feeding access in these instances. PATIENTS AND METHODS: Between May 1993 and December 1999, 21 patients underwent laparoscopic Janeway gastrostomies. Indications were extended incurable tumors of the hypopharnyx (n = 12) and esophagus (n = 9) which rendered oral nutrition impossible and did not allow passage by the endoscope. RESULTS: The mean operative time was 38 minutes (range 24-50). No procedure-related intraoperative or postoperative complications were observed. All patients recovered quickly from surgery and the gastrostomies functioned well in all cases until the death of the patients. CONCLUSION: The laparoscopic Janeway gastrostomy is a safe and simple technique for palliative feeding access, avoiding a laparotomy in patients in whom percutaneous endoscopic placement is impractical. PMID- 11473335 TI - Carcinoma arising in choledochocele. AB - BACKGROUND AND STUDY AIMS: Choledochocele has a potential for carcinogenesis, but no report has described malignant changes of the choledochocele in relation to pancreaticobiliary reflux because its anatomic form does not fit the criteria of pancreaticobiliary malunion (PBM). The aims of this study were to analyze the amylase level in bile in patients with choledochocele and to clarify whether the presence of a choledochocele predisposed to carcinoma. PATIENTS AND METHODS: Records of 2826 patients who had undergone endoscopic retrograde cholangiopancreatography between 1995 and 1999 were reviewed for the presence of choledochocele and/or periampullary carcinoma. As an evidence of pancreaticobiliary reflux, amylase activity was examined in common duct bile obtained at surgery or by endoscopy. The prevalence of periampullary carcinoma was compared between patients with and without choledochocele. RESULTS: A total of 11 patients were diagnosed as having a choledochocele. The amylase level in bile was higher in patients with choledochocele (120,922 +/- 62,269 IU/l; n = 4) than in previously examined patients with functioning gallbladders (15 +/- 24 IU/l; n = 10, P = 0.005). The prevalence of periampullary carcinoma in patients with choledochocele (27%, 3/11) was significantly higher than that in those without choledochocele (0.9%, 26/2815; P<0.0002). CONCLUSION: The bile analysis of the present study presents one possible explanation for the predisposition to carcinoma in choledochocele as bile containing amylase may stagnate in the choledochocele and then carcinoma may develop in the inner epithelium of the choledochocele by the same mechanism as that leading to carcinogenesis in patients with PBM. PMID- 11473336 TI - Endoscopic balloon dilation for removal of bile duct stones: special indications only. PMID- 11473337 TI - Nonsteroidal anti-inflammatory drug-associated colitis with a histology of collagenous colitis. AB - Here we report a case of nonsteroidal anti-inflammatory drug (NSAID)-associated colitis with a histology of collagenous colitis in a 77-year-old woman. The patient had taken aspirin since 1993 after being diagnosed at another hospital, as having multiple cerebral infarctions. She began to suffer from intermittent diarrhea in April 1999. Serological examination showed hypoproteinemia, which indicated that she had protein-losing enteropathy. By July 1999, she had undergone colonoscopic examination four times. Biopsy specimens taken during the fourth colonoscopy revealed collagenous colitis. As the patient had been taking aspirin for 6 years, she was diagnosed as having NSAID-associated colitis with a histology of collagenous colitis. When she stopped taking aspirin, the diarrhea ceased. Three months later, the patient underwent a fifth colonoscopy. A histological examination of the biopsy specimen revealed that the collagen band had vanished. NSAID-associated colitis sometimes shows collagenous colitis histologically and is cured by withdrawing the drug. It is important to differentiate NSAID-associated colitis, even if it shows a histology of collagenous colitis, from collagenous colitis as the two diseases differ in etiology and therapy. PMID- 11473338 TI - Rapid progress of acute suppurative cholangitis to secondary sclerosing cholangitis sequentially followed-up by endoscopic retrograde cholangiography. AB - A 66-year-old man was admitted to our hospital because of right hypochondralgia and fever after colonic polypectomy. Endoscopic examination revealed purulent bile excretion from the duodenal papilla orifice; based on this finding, acute suppurative cholangitis was diagnosed. An endoscopic retrograde cholangiography revealed no abnormality in the biliary tree. However, chronic cholestasis persisted, and endoscopic cholangiography performed 4 months later disclosed a beaded appearance of the intrahepatic bile ducts; this sign is a characteristic finding of sclerosing cholangitis. This is the first report of rapid progression of acute suppurative cholangitis to secondary sclerosing cholangitis sequentially followed-up by endoscopic retrograde cholangiography. PMID- 11473339 TI - Endosonography-guided endoscopic band ligation of Dieulafoy's malformation: a case report. AB - Dieulafoy's vascular malformation may cause serious upper gastrointestinal hemorrhage. The mucosal lesion is usually small, and precise location of the lesion may therefore be difficult. We report a case of Dieulafoy's lesion successfully treated by endoscopic band ligation (EBL) where endoscopic ultrasonography (EUS), including Doppler examination, was applied for accurate diagnosis, guiding EBL and control of treatment. PMID- 11473340 TI - Accidental intrarectal administration of alcohol enema induces proctocolitis and fecal incontinence. PMID- 11473341 TI - An unusual complication following intubation of a benign oesophageal stricture. PMID- 11473342 TI - Images in focus. Peribiliary cysts both in the cystic duct and in the intrahepatic biliary tract. PMID- 11473343 TI - Images in focus. Self-knotting of feeding tube. PMID- 11473344 TI - Lower-than-expected linkage disequilibrium between tightly linked markers in humans suggests a role for gene conversion. AB - Understanding the pattern of linkage disequilibrium (LD) in the human genome is important both for successful implementation of disease-gene mapping approaches and for inferences about human demographic histories. Previous studies have examined LD between loci within single genes or confined genomic regions, which may not be representative of the genome; between loci separated by large distances, where little LD is seen; or in population groups that differ from one study to the next. We measured LD in a large set of locus pairs distributed throughout the genome, with loci within each pair separated by short distances (average 124 bp). Given current models of the history of the human population, nearly all pairs of loci at such short distances would be expected to show complete LD as a consequence of lack of recombination in the short interval. Contrary to this expectation, a significant fraction of pairs showed incomplete LD. A standard model of recombination applied to these data leads to an estimate of effective human population size of 110,000. This estimate is an order of magnitude higher than most estimates based on nucleotide diversity. The most likely explanation of this discrepancy is that gene conversion increases the apparent rate of recombination between nearby loci. PMID- 11473346 TI - Comparative rheological behavior of hyaluronan from bacterial and animal sources with cross-linked hyaluronan (hylan) in aqueous solution. AB - Using a variety of rheological techniques, the behavior of hyaluronan (M(w) 0.8 2.2 x 10(6)), cross-linked hyaluronan (hylan) (M(w) 1.8-12.5 x 10(6)), and Healon (M(w) approximately 5 x 10(6)) (a proprietary hyaluronan) was studied over a large range of molecular weights. The object was to study the effect of the cross links in hylan on the various rheological parameters, in comparison with linear hyaluronan. There are significant differences. The Huggins constant and the critical overlap parameter C*[eta] are considerably lower for hylan and an increase in moduli at low frequencies was observed for hylan compared with the hyaluronan samples at all molecular weights studied. The results point to a difference in structure in dilute solution for hylan due to the ability to form networks, which can be removed by pressure filtration. In contrast, we do not find an increase of the steady shear viscosity and elastic modulus at higher concentrations when a homogeneous entangled network is reached. We attribute this behavior to the semirigid character of the hyaluronan chain and to the predominance of entanglements over the cross-link points present in hylan in the semidilute domain. Due to the higher apparent molecular weights that are possible with hylan structures but not with the hyaluronans currently available, a wider range of applications can be achieved with hylans when viscoelasticity is required, particularly for the viscosupplementation of synovial fluid damaged by osteoarthritis. PMID- 11473345 TI - Paget disease of bone: mapping of two loci at 5q35-qter and 5q31. AB - Paget disease of bone is characterized by focal increases of the bone-remodeling process. It is the second most common metabolic bone disease after osteoporosis. Genetic factors play a major role in the etiology of Paget disease of bone, and two loci have been mapped for the disorder: PDB1 and PDB2. The gene(s) causing the typical form of the disorder remains to be characterized. To decipher the molecular basis of Paget disease of bone, we performed genetic linkage analysis in 24 large French Canadian families (479 individuals) in which the disorder was segregating as an autosomal dominant trait. After exclusion of PDB2, a genomewide scan was performed on the three most informative family nuclei. LOD scores >1.0 were observed at seven locations. The 24 families were then used to detect strong evidence for linkage to chromosome 5q35-qter. Under heterogeneity, a maximum LOD score of 8.58 was obtained at D5S2073, at straight theta= .1. The same characteristic haplotype was carried by all patients in eight families, suggesting a founder effect. A recombination event in a key family confined the disease region within a 6-cM interval between D5S469 and the telomere. The 16 other families, with very low conditional probability of linkage to 5q35-qter, were further used, to map a second locus at 5q31. Under heterogeneity, a maximum LOD score of 3.70 was detected at D5S500 with straight theta=.00. Recombination events refined the 5q31 region within 12.2 cM, between D5S642 and D5S1972. These observations demonstrate the mapping of two novel loci for Paget disease of bone and provide further evidence for genetic heterogeneity of this highly prevalent disorder. It is proposed that the 5q35-qter and 5q31 loci be named "PDB3" and "PDB4," respectively. PMID- 11473347 TI - Molecular dynamics simulation reveals sequence-intrinsic and protein-induced geometrical features of the OL1 DNA operator. AB - We have carried out molecular dynamics simulation of the lambda OL1 DNA operator on the free and the protein-bound forms. Our results lead us to conclude that the binding of the repressor actually makes the N-7 atom of Gua8' more solvent exposed, thereby enhancing its reactivity to chemical methylation. This increase in solvent accessibility surface area occurs simultaneously with the formation of hydrogen bonds between Lys-4 of the nonconsensus flexible N-terminal arm and Gua6' of the nonconsensus half-site operator DNA. Calculations of protein--DNA interaction energies reveal that among the residues of the arm, Lys-4 contributes the most favorably to the interaction energies. This result is consistent with mutagenesis studies that established that lysine at position 4 is absolutely required for tight binding. We find that the nonconsensus arm and the nonconsensus monomer interacts less favorably with DNA than do their respective counterparts of the consensus monomer. Moreover, the six-residue flexible arm accounts for at least half the total protein--DNA interactions energy. These results are in agreement with previous experimental studies. In accord with the diffuse electron density map observed in crystallographic studies of the nonconsensus flexible arm, we find that our model built for this region is more flexible and exhibits more conformations than its consensus counterpart. The simulation also reveals that DNA bending observed near the outer edge of the operator site is an intrinsic sequence-dependent property. By contrast, the DNA bending features observed toward the center of the operator are induced by the protein. On the whole, stepwise protein-induced bending is more pronounced in the consensus half-site operator. We also find that the unusually large helical twist (49 degrees ) observed in the protein-bound form near the center of the operator results from the binding of the protein at a base step with some propensity for high twists. PMID- 11473348 TI - Real-time monitoring of enzymatic hydrolysis of galactomannans. AB - Enzymatic hydrolysis was monitored in real-time using time dependent static light scattering (TDSLS) for a variety of galactomannans from native Brazilian flora. alpha-Galactosidase, which strips only the (1-6)alpha-D galactose side groups, and beta-mannanase, which hydrolyses only the (1-4)beta-D mannan main chain into oligosaccharides were investigated separately and in combination. The time dependent signatures matched those describing side-chain stripping for galactosidase, whereas those resulting from the action of mannanase followed the signature typical of random backbone cleavage. Use of both enzymes together required that the TDSLS theory of polymer degradation be extended to the case where random backbone cleavage sites appear as side chains are stripped by the first enzyme. Whereas galactosidase allowed mannanase to access more backbone cleavage sites as time passes, leading to a higher degree of hydrolysis, there was no increase in rate constants. The distribution of random fragments in the case of mannanase digestion alone followed reasonably well the predictions for random cleavage of a single-strand polymer with a restricted number of cleavage sites. The fragment distributions were evaluated by size exclusion chromatography. PMID- 11473349 TI - Structure and topology of a peptide segment of the 6th transmembrane domain of the Saccharomyces cerevisae alpha-factor receptor in phospholipid bilayers. AB - A detailed analysis of the structure of an 18-residue peptide AQSLLVPSIIFILAYSLK [M6(252-269, C252A)] in 1,2-dimyristoyl-sn-glycero-phosphocholine bilayers was carried out using solid state NMR and attenuated total reflection Fourier transform infrared spectroscopy. The peptide corresponds to a portion of the 6th transmembrane domain of the alpha-factor receptor of Saccharomyces cerevisiae. Ten homologs of M6(252-269, C252A) were synthesized in which individual residues were labeled with (15)N. One- and two-dimensional solid state NMR experiments were used to determine the chemical shifts and (1)H-(15)N dipolar coupling constants for the (15)N-labeled peptides in oriented dimyristoylphosphatidylcholine bilayers on stacked glass plates. These parameters were used to calculate the structure and orientation of M6(252-269, C252A) in the bilayers. The results indicate that the carboxyl terminal residues (9-14) are alpha-helical and oriented with an angle of about 8 degrees with respect to the bilayer normal. Independently, an attenuated total reflection Fourier transform infrared spectroscopy analysis on M6(252-269, C252A) in a 1,2-dimyristoyl-sn glycero-phosphocholine bilayer concluded that the helix tilt angle was about 12.5 degrees. The results on the structure of M6(252-269, C252A) in bilayers are in good agreement with the structure determined in trifluoroethanol/water solutions (B. Arshava et al. Biopolymers, 1998, Vol. 46, pp. 343-357). The present study shows that solid state NMR spectroscopy can provide high resolution information on the structure of transmembrane domains of a G protein-coupled receptor. PMID- 11473350 TI - CD and (13)C(alpha)-NMR studies of folding equilibria in a two-stranded coiled coil formed by residues 190-254 of alpha-tropomyosin. AB - Synthesis and CD and (13)C(alpha)-NMR studies in a near-neutral saline buffer are reported for a 65-residue peptide ((190)Tm(254)) comprising residues 190-254 of the alpha-tropomyosin chain. CD on a version disulfide cross-linked via the N terminal cysteine side chains indicates that this dimer is highly helical and melts near 48 degrees C. The CD is independent of peptide concentration, showing that association of (190)Tm(254) stops at the two-strand level. Similar studies on the reduced version show much lower helix content at low temperature, melting points below room temperature, and the expected concentration dependence. The observed melting temperature of the reduced peptide is far below (by 27 degrees C) that expected from an extant analysis of calorimetry data on parent tropomyosin that designates (190)Tm(254) as an independently melting "cooperative block." This disagreement and the pronounced nonadditivity seen when data for (190)Tm(254) are combined with extant data for other subsequences argue decisively against the concept of specific independently melting blocks within the tropomyosin chain. The data for (190)Tm(254) also serve to test recent ideas on the sequence determinants of structure and stability in coiled coils. Analysis shows that some ideas, such as the stabilizing effect of leucine in the d heptad position, find support, but others--such as the destabilizing effect of alanine in d, the dimer-disfavoring effect of beta-branching in d and its dimer-favoring effect in a, and the dimer-directing effect of asparagine in a--are more questionable in tropomyosin than in the leucine zipper coiled coils. (13)C(alpha) NMR data at two labeled sites, L228(d) and V246(a), of (190)Tm(254) display well separated resonances for folded and unfolded forms at each site, indicating that the transition is slow on the NMR time scale and thus demonstrating the possibility of obtaining thermodynamic and kinetic information on the transition at the residue level. PMID- 11473351 TI - (13)C cross-polarization/magic angle spinning NMR studies of alpha-elastin preparations show retention of overall structure and reduction of mobility with a decreased number of cross-links. AB - High-resolution solid-state (13)C NMR spectra are presented for samples of alpha elastin prepared from the aorta of normal and copper-deficient pigs. Chemical shifts of the various peaks indicate that both the normal and undercross-linked peptides have similar overall structures. However, (13)C T(1), (13)C T(1 rho), and (1)H T(1 rho) measurements indicate that the alpha-elastin peptides obtained from the abnormal elastic fibers samples exhibit altered mobilities, particularly in their side chains. Results from spectra taken with a range of contact times and from dipolar dephasing experiments are consistent with conclusions reached with the relaxation measurements. Namely, the loss of function associated with the undercross-linked sample is correlated to a small but measurable difference in relative mobility. PMID- 11473352 TI - Effects of hydrogen-bond deletion on peptide helices: structural characterization of depsipeptides containing lactic acid. AB - The insertion of alpha-hydroxy acids into peptide chains provides a convenient means for investigating the effects of hydrogen bond deletion on polypeptide secondary structures. The crystal structures of three oligopeptides containing L lactic acid (Lac) residue have been determined. Peptide 1, Boc-Val-Ala-Leu-Aib Val-Lac-Leu-Aib-Val-Ala-Leu-OMe (Boc: tert-butyloxycarbonyl; Aib: alpha- aminoisobutyric acid; OMe: methyl ester), and peptide 2, Boc-Val-Ala-Leu-Aib-Val Lac-Leu-Aib-Val-Leu-OMe, adopt completely helical conformations in the crystalline state with the Lac(6) residue comfortably accommodated in the center of a helix. The distance between the O atoms of Leu(3) CO group and the Lac(6) O (ester) in both the structures is 3.1-3.3 A. The NMR and CD studies of peptide 1 and its all-amide analogue 4, Boc-Val-Ala-Leu-Aib-Val-Ala-Leu-Aib-Val-Ala-Leu OMe, provide firm evidence for a continuous helical conformation in solution in both the cases. In a 14-residue peptide 3, Boc-Val-Ala-Leu-Aib-Val-Ala-Leu-Val Ala-Leu-Aib-Val-Lac-Leu-OMe, residues Val(1)-Leu(10) adopt a helical conformation. Aib(11) is the site of chiral reversal resulting in helix termination by formation of a Schellman motif. Residues 12-14 adopt nonhelical conformations. The loss of the hydrogen bond near the C-terminus appears to facilitate the chiral reversal at Aib(11). Published 2001 John Wiley & Sons, Inc. Biopolymers 59: 276-289, 2001 PMID- 11473353 TI - Applications of fluorescence correlation spectroscopy: measurement of size-mass relationship of native and denatured schizophyllan. AB - Diffusion dynamics of a polysaccharide, schizophyllan has been studied by fluorescence correlation spectroscopy (FCS). Several different sizes of nondenatured and denatured schizophyllan have been labeled with rhodamine 6G in borate buffer. The length of the nondenatured schizophyllan was calculated from FCS data by using the Broersma's relationship for rod-like macromolecules. The obtained length was close to that obtained by atomic force microscopy (AFM) measurements. Denatured schizophyllan possesses a random coil conformation. Its hydrodynamic radius R(h) was measured by FCS. The relationship between R(h) and the molecular mass M has been studied and the scaling relationship R(h)--M(0.59) has been obtained, which is in agreement with the random coil model with excluded volume effect. The persistence length q(denat) of the denatured schizophyllan was determined by Hearst's relationship, to be equal to 5.16 +/- 0.75 (nm). The work demonstrates the utility of FCS method for dynamics investigations of biopolymers especially in diluted regime (concentration lower than 10(-8)M could be measured) where other techniques could not be used. PMID- 11473354 TI - Hsp90: chaperoning signal transduction. AB - Hsp90 is an ATP dependent molecular chaperone involved in the folding and activation of an unknown number of substrate proteins. These substrate proteins include protein kinases and transcription factors. Consistent with this task, Hsp90 is an essential protein in all eucaryotes. The interaction of Hsp90 with its substrate proteins involves the transient formation of multiprotein complexes with a set of highly conserved partner proteins. The specific function of each component in the processing of substrates is still unknown. Large ATP-dependent conformational changes of Hsp90 occur during the hydrolysis reaction and these changes are thought to drive the chaperone cycle. Natural inhibitors of the ATPase activity, like geldanamycin and radicicol, block the processing of Hsp90 substrate proteins. As many of these substrates are critical elements in signal transduction, Hsp90 seems to introduce an additional level of regulation. PMID- 11473355 TI - Kinetics of endomitosis in primary murine megakaryocytes. AB - Megakaryocytes (MKs) develop from diploid progenitor cells via successive rounds of DNA synthesis in the absence of cell division, a process termed endomitosis (EnM). While the mechanism underlying EnM is not known, studies in yeast and leukemic cell lines have suggested that it may be due to reduced levels of cyclin B1 or cdc2, leading to a decrease in mitotic kinase activity. Using flow cytometry to study EnM highly purified marrow-derived MK precursors, we found that: (1) on average, 36% of 8N-32N MKs expressed abundant cyclin B during G2/M. The percentage of cells in G2/M decreased in >64N MKs, suggesting the limit of EnM, (2) the level of cyclin B per G2/M MK increased linearly with ploidy, (3) cyclin B expression oscillated normally in polyploid MKs, (4) MPM-2, a phosphoepitope created by the action of mitotic kinases and specific to M-phase cells, was expressed in a significant fraction of polyploid MKs, and (5) there was an apparent increase of cyclin B in G1-phase in polyploid MKs. This study provides the first qualitative kinetic data regarding the cell cycle status of MKs within individual ploidy classes. It also demonstrates the feasibility of using anti-cyclin B antibody and flow cytometry to resolve G1 from G2/M populations in polyploid MKs. Finally, these findings establish that neither a relative nor absolute deficiency of mitotic kinase components is responsible for EnM, suggesting that the departure from normal cell division kinetics seen in polyploid MKs is likely due to alterations in other cell cycle regulators. PMID- 11473356 TI - Endothelial exposure to hypoxia induces Egr-1 expression involving PKCalpha mediated Ras/Raf-1/ERK1/2 pathway. AB - Hypoxia induces endothelial dysfunction that results in a series of cardiovascular injuries. Early growth response-1 (Egr-1) has been indicated as a common theme in vascular injury. Here we demonstrates that in bovine aortic endothelial cells (ECs) subjected to hypoxia (PO(2) approximately 23 mmHg), rapidly increased Egr-1 mRNA expression which peaked within 30 min and decreased afterwards. Treatment of ECs with PD98059, a specific inhibitor to mitogen activated protein kinase (MAPK/ERK), inhibited this hypoxia-induced Egr-1 expression. The involvement of ERK pathway was further substantiated by the inhibition of Egr-1 promoter activities when ECs were co-transfected with a dominant negative mutant of Ras (RasN17), Raf-1 (Raf 301), or a catalytically inactive mutant of ERK2 (mERK). In addition, the hypoxia-induced transcriptional activity of Elk-1, an ERK substrate, was abolished by administration of PD98059. Addition of calphostin C, a protein kinase C (PKC) inhibitor, completely blocked the hypoxia-augmented Egr-1 expression. The likewise occurred while exposing ECs to D609 to inhibit phospholipase C and BAPTA/AM to chelate intracellular calcium. Hypoxia to ECs increased ERK phosphorylation within 10 min and which was abolished by administration of PD98095, calphostin C, and BAPTA/AM. Hypoxia triggered a transient translocation of PKCalpha from cytosol to membrane fraction concurrent with the association of PKCalpha to Raf-1. Involvement of PKCalpha in mediating ERK activation was further confirmed by the inhibition of ERK and the subsequent Egr-1 gene induction with antisense oligonucleotides to PKCalpha. These results indicate that ECs under hypoxia induce Egr-1 expression and this induction requires calcium, phospholipase C activation, and PKCalpha-mediated Ras/Raf-1/ERK1/2 signaling pathway. Our finding support the importance of specific PKC isozyme linked to MAPK pathway in the regulation of endothelial responses to hypoxia. PMID- 11473357 TI - Suppression of inducible nitric oxide generation by agmatine aldehyde: beneficial effects in sepsis. AB - The induction of inducible nitric oxide synthase (iNOS) serves an important immuno-protective function in inflammatory states, but ungoverned nitric oxide (NO) generation can contribute to a number of pathologic consequences. Delineation of the mechanisms that can downregulate iNOS-generated NO in inflammation could have therapeutic relevance. Here we show that agmatine, a metabolite of arginine, inhibits iNOS mediated nitric oxide generation in cytokine stimulated cell culture preparations. This effect was not cell type specific. Increased diamine oxidase (DAO) and decreased aldehyde dehydrogenase (AldDH) activities are also representative of inflammatory settings. Increasing the conversion of agmatine to an aldehyde form by addition of purified DAO or suppression of aldehyde breakdown by inhibition of AldDH activity increases the inhibitory effects of agmatine in an additive fashion. Inhibitors of DAO, but not monoamine oxidase (MAO), decreased the inhibitory effects of agmatine, as did the addition of AldDH or reacting aldehydes with phenylhydrazine. We examined rats given lipopolysaccharide (LPS) to evaluate the potential effects of agmatine in vivo. Endotoxic rats administered agmatine prevented the decreases in blood pressure and renal function normally associated with sepsis. Agmatine treatment also increased the survival of LPS treated mice. Our data demonstrate the capacity of agmatine aldehyde to suppress iNOS mediated NO generation, and indicate a protective function of agmatine in a model of endotoxic shock. How agmatine may aid in coordinating the early NO phase and the later repair phase responses in models of inflammation is discussed. PMID- 11473358 TI - Cytosolic phospholipase A(2) activity during the ongoing cell cycle. AB - Cytosolic phospholipase A(2) (cPLA(2)) is of special interest because it selectively releases arachidonic acid from membrane phospholipids. Arachidonic acid has been implicated to play an important role in various cellular responses. Recently arachidonic acid release and prostaglandin synthesis have been shown to be cell cycle dependent and therefore the activity of cPLA(2) during the ongoing cell cycle was investigated, using the mitotic shake off method for cell synchronisation. cPLA(2) activity was high in mitotic cells and decreased rapidly in the early G1 phase. A strong increase in activity was measured following the G1/S transition in both neuroblastoma and Chinese hamster ovary cells. The changes in activity were not due to a difference in cPLA(2) expression but due to phosphorylation of cPLA(2). Phosphorylation of cPLA(2) occurs through MAPK since the use of a specific MAPK kinase inhibitor and serum depletion of synchronised cells inhibited cPLA(2) activity. PMID- 11473359 TI - Functional characterization of P2Y and P2X receptors in human eosinophils. AB - Activation of purinoceptor by ATP induces in eosinophils various cell responses including calcium transients, actin polymerization, production of reactive oxygen metabolites, CD11b-expression, and chemotaxis. Here, the effect of ion channel gated P2X and/or G protein-coupled P2Y receptor agonists ATP, ATPgammaS, alpha,beta-meATP, 2-MeSATP, BzATP, ADP, CTP, and UTP on the intracellular Ca(2+) mobilization, actin polymerization, production of reactive oxygen metabolites, CD11b expression and chemotaxis of human eosinophils were measured and the biological activity was analyzed. Although all tested nucleotides were able to induce all these cell responses, the biological activity of the analyzed nucleotides were distinct. Agonists of the G protein-coupled P2Y receptors such as 2-MeSATP, UTP, and ADP have a higher biological activity for production of reactive oxygen metabolites, actin polymerization and chemotaxis in comparison to the ion channel-gated P2X agonists alphabeta-meATP, BzATP, and CTP. In contrast, P2Y and P2X agonist showed similar potencies in respect to intracellular calcium transient and CD11b up-regulation. This conclusion was further supported by experiments with receptor iso-type antagonist KN62, EGTA or with the G(i) protein inactivating pertussis toxin. These findings indicate participation of different purinorecptors in the regulation of cell responses in eosinophils. PMID- 11473360 TI - Effects of nitric oxide and peroxynitrite on endotoxin-induced leukocyte adhesion to endothelium. AB - Leukocyte accumulation has been shown to be increased in sepsis. Moreover, in inducible nitric oxide synthase (iNOS) knockout mice, a further increase in leukocyte accumulation has been observed during sepsis, suggesting that nitric oxide (NO) may affect leukocyte/endothelial interaction. Accelerated peroxynitrite formation also occurs during sepsis. In the present study, the effect of peroxynitrite or NO on leukocyte adhesion to nitric oxide synthase (NOS)-inhibited or endotoxin-treated endothelium was examined. Bovine aortic endothelial cells were treated with either L-NAME or lipopolysaccharide (LPS) and interferon-gamma for 4 hr and subsequent leukocyte adhesion was measured. Both L NAME and LPS treatment resulted in increased leukocyte adhesion compared with control. Neither a peroxynitrite donor, SIN-1, nor a direct NO donor, DETA-NO, had any effect on leukocyte adhesion to untreated endothelium. However, when the L-NAME or LPS-treated endothelial cells were treated simultaneously with either SIN-1 or DETA-NO, there was a significant reduction in leukocyte adhesion. Moreover, at the concentrations used in the present study, neither peroxynitrite nor NO showed harmful effects on normal cultured endothelial cells. These data demonstrating inhibition of leukocyte adhesion to endotoxin-treated endothelium suggest that peroxynitrite or NO may exert a beneficial effect during sepsis. PMID- 11473361 TI - PTHrP expression in chick sternal chondrocytes is regulated by TGF-beta through Smad-mediated signaling. AB - PTHrP regulates the rate of chondrocyte differentiation during endochondral bone formation. The expression of PTHrP and its regulation by TGF-beta, BMP-2, and PTHrP was examined in upper sternal chondrocytes following 1, 3, and 5 days of continuous treatment. While TGF-beta stimulated the expression of PTHrP (5-fold), PTHrP caused a slight inhibition, and BMP-2 markedly inhibited PTHrP mRNA expression. The effect of these factors on PTHrP expression was not simply related to the maturational state of the cells, since BMP-2 increased, while both PTHrP and TGF-beta decreased the expression of type X collagen. TGF-beta isoforms 1, 2, and 3 all stimulated PTHrP expression. Signaling events involved in the induction of PTHrP by TGF-beta were further evaluated in a PTHrP-promoter CAT construct. The effect of TGF-beta, BMP-2, and PTHrP on the PTHrP-promoter paralleled their effects on mRNA expression, with TGF-beta significantly increasing CAT activity, BMP-2 decreasing CAT activity, and PTHrP having a minimal effect. Co-transfection of the TGF-beta signaling molecule, Smad 3, mimicked the effect of TGF-beta (induction of PTHrP promoter), while dominant negative Smad 3 inhibited the induction of the PTHrP promoter by TGF-beta. Furthermore, infection with a Smad 3-expressing retrovirus mimicked the effects of exogenously added TGF-beta, and induced PTHrP mRNA expression in the infected chondrocyte culture. In contrast, a dominant negative Smad 3 completely inhibited PTHrP promoter stimulation by TGF-beta, but only partially blocked the effect of TGF-beta on PTHrP mRNA synthesis. These findings demonstrate that PTHrP is expressed in chondrocytes undergoing endochondral ossification, and show regulation, at least in part, by TGF-beta through Smad mediated signaling events. PMID- 11473362 TI - Involvement of transcriptional repressor ATF3 in acceleration of caspase protease activation during DNA damaging agent-induced apoptosis. AB - DNA topoisomerase inhibitors are effective chemotherapeutic agents on several solid tumor cells. They induce a specific signaling cascade that executes an active cell death process (apoptosis), including caspase activation, and the blockage of the signaling is associated with drug-resistance of tumor cells. However, little is known about the initial signal transduction induced by the agents. In the present study, we screened genes that are initially upregulated in caspase-independent manner. We found that the activating transcription factor 3 (ATF3) protein, a repressor of cyclic-AMP responsive element (CRE)-dependent transcription, was strongly induced among CRE-BP/ATF members and subsequently accumulated in nuclei following camptothecin or etoposide treatment. During induction of apoptosis, the accumulation and the nuclear translocation of ATF3 coincided with the activation of caspase protease and were not inhibited by the broad caspase inhibitor Z-VAD-fmk, indicating that ATF3 induction is not a downstream event of caspase activation. When stably or transiently overexpressed, ATF3 markedly accelerated the drug-induced apoptosis and enhanced caspase protease activation. ATF3 strongly downregulated CRE-dependent transcription, while ATF3 did not affect the expression levels of Bcl-2, Bcl-x, or Bax. Our present results indicate that ATF3 plays a critical role in accelerating caspase protease activation and apoptosis. Since CRE-dependent transcription functions as cell survival signaling, ATF3 could control the upstream signaling of apoptosis by repressing CRE-dependent gene expression of cell survival factors. PMID- 11473363 TI - Expression of vascular endothelial growth factor receptors in smooth muscle cells. AB - Vascular Endothelial Growth Factor (VEGF) has been typically considered to be an endothelial-specific growth factor. However, it was recently demonstrated that VEGF can interact with non endothelial cells. In this study, we tested whether vascular smooth muscles cells (VSMCs) can express VEGF receptors, such as flk-1, flt-1, and neuropilin (NP)-1, and respond to VEGF in vitro. In cultured VSMCs, flk-1 and flt-1 expression was inversely related to cell density. The expression of flk-1 was down-regulated with increasing passage numbers. However, NP-1 levels were not affected by cell density or passage numbers. Flk-1, Flt-1, and NP-1 protein levels were confirmed by Western Blotting. Although the functional mature form of Flk-1 protein is expressed at low levels in VSMCs, phosphorylation of Flk 1 following VEGF(165) stimulation was still observed. SMCs migrated significantly in response to VEGF(165) and VEGF-E, whereas Placenta Growth Factor (PlGF) induced migration only at higher concentrations. Since VEGF-E is a specific activator of flk-1 while PlGF specifically activates only flt-1, SMC migration induced by VEGF(165) is likely to be mediated primarily through the flk-1 receptor. VSMCs did not significantly proliferate in response to VEGF(165), PlGF, and VEGF-E. In conclusion, our studies demonstrate the presence of VEGF receptors on VSMCs that are functional. These studies also indicate that in vivo, VEGF may play a role in modulating the response of VSMCs. PMID- 11473364 TI - Role of phosphoinositide 3OH-kinase in autocrine transformation by PDGF-BB. AB - Phosphoinositide 3OH-kinases (PI3K) are a family of lipid kinases that activates signalling pathways important for migration, cytoskeletal rearrangements, and cell survival. These processes are important hallmarks in transformation. We have evaluated the functional role of PI3K for development of a transformed morphology and migratory responses of murine fibroblasts (NIH/sis and COL1A1/NIH3T3 cell lines) stimulated in an autocrine fashion by constitutive expression of platelet derived growth factor-BB (PDGF-BB). We show that prolonged treatment with the specific PI3K inhibitor LY294002, induced a reversion of the transformed morphology, and prevented density-independent growth and focus formation. Functional PI3K was also required for development of the transformed morphology of NIH/sis and COL1A1/NIH3T3. Furthermore, treatment with LY294002 completely perturbed random migration of the cells. In addition our data show that, in the signalling pathways downstream of PI3K, activation of the small GTPase Rac was a prerequisite for the transformation signal. Our data also indicate the presence of a suramin-insensitive PI3K activity. Most likely this was due to the presence of a suramin-insensitive intracellular PDGFR pool that allowed activation of PI3K located in intracellular compartments. In conclusion these data show that intact PI3K activity was required for the morphological alterations and the enhanced migratory response that are hallmarks for PDGF induced autocrine transformation. PMID- 11473365 TI - Autocrine TGFbeta signaling mediates vitamin D3 analog-induced growth inhibition in breast cells. AB - In this study, we address whether TGFbeta signaling mediates vitamin D3 analog induced growth inhibition in nonmalignant and malignant breast cells. Normal mammary epithelial cells (184), immortalized nonmalignant mammary epithelial cells (184A1 and MCF10A), and breast cancer cells (early passage MCF7: MCF7E) were sensitive to the inhibitory effects of vitamin D3 analogs (EB1089 and MC1288) while late passage MCF7 breast cancer (MCF7L) cells were relatively resistant. A similar pattern of sensitivity to TGFbeta was observed with these cells. Thus, the sensitivity to the vitamin D3 analogs correlated with the sensitivity to TGFbeta. MCF7L TGFbetaRII-transfected cells, which have autocrine TGFbeta activity, were more sensitive to EB1089 than MCF7L cells. TGFbeta neutralizing antibody was found to block the inhibitory effects of these analogs. These results are consistent with the idea that autocrine TGFbeta signaling mediates the anti-proliferative effects of the vitamin D3 analogs in these cells. The expression of TGFbeta isoforms and/or TGFbeta receptors was induced by the analogs in the vitamin D3 and TGFbeta sensitive cells. Vitamin D3 analogs did not induce TGFbeta or TGFbeta receptor expression in the resistant MCF7L cells. Therefore, EB1089 induces autocrine TGFbeta activity through increasing expression of TGFbeta isoforms and/or TGFbeta receptors. In addition, EB1089 induced nuclear VDR protein levels in the sensitive 184A1 cells but not in the resistant MCF7L cells. 184A1 cells were more sensitive to EB1089-induced VDR dependent transactivation than MCF7L cells as measured by a luciferase reporter construct containing the VDRE, indicating a defect of VDR signaling in MCF7L cells. Smad3, a TGFbeta signaling mediator, coactivated VDR-dependent transactivation in 184A1 cells but not in MCF7L cells. These results indicate that Smad3 coactivates VDR to further enhance TGFbeta signaling and vitamin D3 signaling in the sensitive 184A1 cells. The results also indicate that Smad3 is not of itself sufficient to coactivate VDR in TGFbeta/vitamin D3 resistant MCF7L cells and other factors are required. We found that the PI 3-kinase pathway inhibitor LY29004 inhibited the synergy of TGFbeta and EB1089 on VDR-dependent transactivation activity. This indicates that the crosstalk between TGFbeta and vitamin D signaling is also PI 3-kinase pathway dependent. PMID- 11473366 TI - Vascular smooth muscle cell growth-promoting factor/F-spondin inhibits angiogenesis via the blockade of integrin alphavbeta3 on vascular endothelial cells. AB - Vascular smooth muscle cell growth-promoting factor (VSGP) was originally isolated from bovine ovarian follicular fluid as a stimulator of vascular smooth muscle cell proliferation. Homology searches indicate that bovine and human VSGPs are orthologs of rat F-spondin. Here, we examined whether recombinant human VSGP/F-spondin affected the biological activities of endothelial cells. VSGP/F spondin did not affect the proliferation of human umbilical vein endothelial cells (HUVECs); however, it did inhibit VEGF- or bFGF-stimulated HUVEC migration. To clarify the mechanism of this inhibitory effect, we examined the adhesion of HUVECs to extracellular matrix proteins. VSGP/F-spondin specifically inhibited the spreading of HUVECs on vitronectin via the functional blockade of integrin alphavbeta3. As a result, VSGP/F-spondin inhibited the tyrosine phosphorylation of focal adhesion kinase (FAK) when HUVECs were plated on vitronectin. Moreover, VSGP/F-spondin inhibited the activation of Akt when HUVECs on vitronectin were stimulated with VEGF. VSGP/F-spondin inhibited tube formation by HUVECs in vitro and neovascularization in the rat cornea in vivo. These results indicate that VSGP/F-spondin inhibits angiogenesis at least in part by the blockade of endothelial integrin alphavbeta3. PMID- 11473367 TI - Monocytic cell adhesion to intact and plasmin-modified fibrinogen: possible involvement of Mac-1 (CD11b/CD18) and ICAM-1 (CD54). AB - beta(2)-integrin Mac-1 and immunoglobulin-like ICAM-1 adhesion molecules are expressed by monocytes and both known to bind fibrinogen and its degradation products. Here, we investigated whether fibrinogen cleavage with plasmin modulates the adherence of monocytic cells and what types of adhesion molecules are involved. Using several cell types, characterized by different patterns of Mac-1 and ICAM-1 expression, and monoclonal antibodies against beta(2)-integrins and ICAM-1 we demonstrate, that fibrinogen cleavage evokes gradual decrease in beta(2)-integrin-dependent cell adhesion. Furthermore, generation of the early degradation products, fragments X and Y, by minimum cleavage of fibrinogen stimulates cell adhesion, mediated by ICAM-1. PMID- 11473368 TI - Radical versus conservative cancer surgery: an anachronism? PMID- 11473369 TI - Breast cancer in the third millennium. PMID- 11473370 TI - AJCC/TNM cancer staging, present and future. PMID- 11473371 TI - FDG-PET sensitivity for melanoma lymph node metastases is dependent on tumor volume. AB - BACKGROUND AND OBJECTIVES: The purpose of this study is to determine the tumor volume threshold for successful positron emission tomography (PET) imaging of melanoma nodal metastases. METHODS: Review of a clinical series of patients who had FDG-PET imaging of regional lymph node basins followed by lymphadenectomy. Lymph node tumor volumes were calculated from direct measurements of metastatic nodule(s) in formalin fixed specimens. PET scan interpretations were correlated with histology to determine sensitivity. Sensitivity was correlated with the aggregate lymph node tumor volume in the nodal basin and with AJCC stage group. RESULTS: Forty-five patients with 49 pathologically positive regional nodal basins comprised the study group. Median total basin tumor volume was 28.3 mm(3)(range 0.004-22,879 mm(3)). FDG-PET sensitivity for detection of all tumor volumes was 0.49. The observed 90% sensitivity threshold for detection of nodal metastases was > or = 78 mm(3). PET sensitivity was 0.14 for detection of tumor volumes < 78 mm(3). PET sensitivity differed by prescan AJCC stage: I-0.0; II 0.24; III-0.81; IV-1.0 (P < 0.001). CONCLUSIONS: FDG-PET reliably detects lymph node tumor deposits greater than approximately 80 mm(3) volume, but sensitivity falls rapidly below this. This amount of tumor is most likely to occur in patients with AJCC stage III or IV disease. PMID- 11473372 TI - Accuracy of sentinel node biopsy in predicting nodal status in patients with breast carcinoma. AB - BACKGROUND AND OBJECTIVES: While sentinel lymph node biopsy is considered by many to have replaced axillary node dissection in the management of breast cancer, concerns remain regarding false-negative results. METHODS: To investigate the accuracy of sentinel node biopsy, we reexamined all sentinel and nonsentinel nodes with multilevel sectioning and immunohistochemical staining in 42 consecutive cases of breast cancer in which sentinel node biopsy was performed and followed by axillary dissection. RESULTS: By routine hematoxylin and eosin (H&E) staining, 34% of patients were found to be node positive, with no cases of false-negative sentinel node biopsy. Reevaluation of 775 negative sentinel and nonsentinel nodes with an additional two levels and immunohistochemistry identified three "node-negative" patients who had micrometastases in the sentinel node, increasing detection in 8% of cases. More important, is the fact however, that there were no cases where additional sections and immunohistochemistry identified metastases in nonsentinel nodes that had bypassed the sentinel node. The accuracy of the sentinel node in predicting the nodal status was 100%. CONCLUSIONS: Cytokeratin immunohistochemistry will identify more patients with nodal micrometastases; however, it was unable to identify any cases where micrometastases were present in nonsentinel nodes when the sentinel node was negative. The status of the sentinel node accurately identifies the status of the axillary basin. PMID- 11473373 TI - Folate status among patients with non-small cell lung cancer: a case-control study. AB - BACKGROUND AND OBJECTIVES: This case control study explored the purported inverse relationship between folate status and lung cancer development. METHODS: Folate status of 46 postoperative non-small cell lung cancer (NSCLC) patients was compared to that of 44 non-cancer patients. Cancer patients had completed treatment > 3 months prior and had no evidence of cancer. Ineligibility criteria for all patients included (1) > 2 alcoholic drinks/day (2) ongoing tobacco use, or (3) folate supplementation > 400 microg/day. RESULTS: No differences were found between groups in serum and RBC folate after adjustment for age and use of folate-interfering medications: geometric means (GM) x /geometric standard error (GSE): 7.9 ng/ml x /1.1 vs. 7.8 ng/ml x /1.1, respectively (P = 0.91) for serum folate; 264 ng/ml x /1.1 vs. 263 ng/ml x /1.1, respectively (P = 0.97) for RBC folate. Age- and creatinine-adjusted homocysteine was no different between groups: GM x /GSE: 9.4 micromol/L x /1.0 vs. 8.6 micromol/L x /1.0, respectively (P = 0.17). No difference were seen in folate intake. Frequencies of the homozygous genotype for the MTHFR polymorphism, an enzyme important in folate metabolism and associated with a reduced risk of other cancers, were no different. CONCLUSIONS: This case control study does not support the hypothesis that low folate is an independent risk factor for NSCLC. PMID- 11473374 TI - Optical coherence tomography monitoring for laser surgery of laryngeal carcinoma. AB - BACKGROUND AND OBJECTIVES: The goal of this study is to apply a new bioimaging modality, the Optical Coherence Tomography (OCT), for intraoperative control in laser surgery of laryngeal carcinoma. STUDY DESIGN/MATERIALS AND METHODS: We studied 26 patients with laryngeal carcinoma in situ and in T(1), T(2) stage. We used an endoscopic OCT device for imaging at a wavelength of 0.83 microm with the acquisition rate of approximately 0.5 frames/s for a single (200 x 200 pixel) tomogram. All patients were operated with a surgical YAG:Nd laser at two switchable wavelengths of 1.44 microm and 1.32 microm by laryngofissure, direct microlaryngoscopy, and fibrolaryngoscopy. RESULTS: Information on structural alterations in laryngeal mucosa to the depth of 2 mm, obtained by OCT, makes it possible to precisely locate tumor borders, thus giving an opportunity to control the surgical treatment of laryngeal carcinoma. The YAG:Nd laser scalpel with wavelengths of 1.32 microm and 1.44 microm is successful in surgical procedures both in open and closed larynx due to efficient coagulation and minimization of collateral tissue damage area. Combination of the two wavelengths in the single laser unit and intraoperative OCT monitoring result is a new modality for minimally invasive larynx surgery. CONCLUSIONS: OCT is promising to become a new diagnosing method of laryngeal carcinoma and a tool for laser treatment monitoring. PMID- 11473375 TI - Status of hyperthermia in the treatment of advanced liver cancer. AB - The vast majority of patients with malignant liver tumors have inoperable disease. These patients must rely on chemotherapy, radiotherapy, and various locoregional treatments. Although these treatments have demonstrated encouraging response rates, symptom palliation and occasional down staging of tumors, their impact on survival is minor. As a result there has been renewed interest in hyperthermia as a treatment option. This study reviews the current modalities of hyperthermia in terms of clinical results, side effects, limitations, and therapeutic standing. PMID- 11473376 TI - Quality of life as an outcome in clinical trials and cancer care: a primer for surgeons. PMID- 11473377 TI - Intraoperative needle decompression: a simple alternative to nasogastric decompression. PMID- 11473378 TI - Laparoscopy of the right iliac fossa through a paramedian incision. PMID- 11473379 TI - Is the groin region a good source for full-thickness skin graft? PMID- 11473380 TI - Mucinous colorectal carcinoma. PMID- 11473382 TI - DNA damage in human transitional cell carcinoma cells after exposure to the proximate metabolite of the bladder carcinogen 4-aminobiphenyl. AB - The DNA damage induced by N-hydroxy-4-acetylaminobiphenyl (N-OH-AABP), the proximate carcinogenic metabolite of the human bladder carcinogen 4-aminobiphenyl (ABP), was examined in human transitional cell carcinoma (TCC) cells after exposure to the chemical in vitro. 32P-postlabeling analysis of TCC cultures exposed to N-OH-AABP revealed a minor adduct identified as 3-(deoxyguanosin-N2 yl)-4-acetylaminobiphenyl (dG-N2-AABP) based on comparison of the HPLC and TLC mobility of the product with the synthetic standard. An adduct with the same chromatographic properties was also detected on postlabeling analyses of calf thymus DNA bound to N-OH-AABP by incubation with horseradish peroxidase and hydrogen peroxide. Detection of dG-N2-AABP, which contains the acetyl moiety, suggests that N-acetoxy-4-acetylamino-biphenyl might be formed as a reactive intermediate and could conceivably arise by a free-radical-mediated reaction of N OH-AABP with endogenous peroxidases. The radical intermediates could also form reactive oxygen species (ROS). To test this possibility, TCC cultures were exposed to N-OH-AABP and the formation of ROS was measured using 2,7 dichlorofluorescein (DCF) fluorescence assay. TCC cultures exposed to N-OH-AABP showed a dose-dependent increase in the ratio of DCF/DNA fluorescence compared to the untreated controls. Formation of ROS was inhibited by butylated hydroxyanisole (BHA). Furthermore, oxidative DNA damage resulting from ROS was monitored by measurement of 8-oxoguanine products by immunochemical staining and the TCC cells treated with N-OH-AABP revealed a characteristic staining. These results suggest that N-OH-AABP caused oxidative DNA damage as well as bulky covalent adducts in urothelial DNA, possibly involving endogenous peroxidases. These findings show that human uroepithelial cells, which are the target cell types in vivo for arylamine-induced cancers, are metabolically capable of activating these proximate carcinogenic metabolites of arylamines, and these reactions might play a determinate role in the genotoxicity of these environmental carcinogens. PMID- 11473383 TI - Recombinant human P450 forms 1A1, 1A2, and 1B1 catalyze the bioactivation of heterocyclic amine mutagens in Escherichia coli lacZ strains. AB - Three recombinant human P450 enzymes, forms 1A1, 1A2, and 1B1, were coexpressed with NADPH-cytochrome P450 reductase in an E. coli lacZ strain suitable for detection of the mutagenicity of heterocyclic and aromatic amines. The resulting strains expressed the recombinant P450 holoenzymes at high levels. MeIQ (2-amino 3,4-dimethylimidazo[4,5-f]quinoline) was activated effectively by P450 1A2, weakly by P450 1A1, and not detectably by P450 1B1. MeIQx (2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline) and Trp-P-2 (3-amino-1-methyl-5H-pyrido[4,3 b]indole) were activated by all three enzymes, with form 1A2 the most effective. These strains facilitate analysis of the substrate specificity of human P450 forms that participate in the metabolic activation of carcinogens. PMID- 11473384 TI - A single (6-4) photoproduct inhibits plasmid DNA replication in xeroderma pigmentosum variant cell extracts. AB - The human skin cancer-prone disease xeroderma pigmentosum variant (XPV) results from a mutation in the human RAD30 gene, which encodes the lesion bypass DNA polymerase eta. XPV cells are characterized by delayed completion of DNA replication and increased mutagenesis following UV-irradiation. Using extracts of an XPV lymphoblast cell line (GM2449C) that has a truncating mutation in the RAD30 gene, we investigated the effect of a (6-4) photoproduct and a cyclobutane pyrimidine dimer (CPD), at a unique -TT- site on either the leading or lagging strand, on plasmid DNA replication. Compared to normal cell extracts, XPV cell extracts have a reduced capacity to carry out complete replication of DNA containing either a (6-4) photoproduct or a CPD on the leading strand, whereas there is little difference between the two cell extracts in replication of DNA containing a lesion on the lagging strand. Inhibition of replication in the presence of a (6-4) photoproduct is attributed to arrest of nascent DNA strand synthesis at the lesion site; in XPV cell extracts, the proportion of arrested products is increased compared to that of normal cell extracts. These results are consistent with a requirement for functional DNA polymerase eta in the replication of a double-stranded plasmid containing either a (6-4) photoproduct or a CPD, on the leading but not the lagging strand. PMID- 11473385 TI - Transplacental mutagenicity of N-ethyl-N-nitrosourea at the hprt locus in T lymphocytes of exposed B6C3F1 mice. AB - Previous studies have compared age-related differences in total mutagenic burden in mice of differing age (preweanling, weanling, or young adult) after single intraperitoneal (i.p.) injections of ethylnitrosourea (ENU). The purpose of the present investigation was to determine the effects of time elapsed since treatment on the frequency of hprt mutant T-cells (Mf) from mice treated transplacentally with single acute vs. multiple split doses of ENU. To this end, pregnant C57BL/6 mice (n = 13-16/group), which had been bred to C3H males, were given i.p. injections of 40 mg ENU/kg bw in a single dose on day 18 of gestation, in a split dose of 6 mg ENU/kg bw on days 12 through 18 of gestation, or DMSO vehicle alone. Groups of pups were necropsied on days 10, 13, 15 (single dose only), 17, 20, 40, and 70 postpartum for T-cell isolations and hprt Mf measurements using the T-cell cloning assay. The time required to reach maximum Mfs in T-cells isolated from thymus of transplacentally treated animals was 2 weeks, the same time span as previously observed after ENU treatment of adult, weanling, and preweanling mice. Mfs in T-cells isolated from spleens of control animals averaged 2.1 +/- 0.3 (SE) x 10(-6). In spleens of mice treated transplacentally with ENU in a single dose, Mfs reached a maximum at 15 days postpartum [84.7 +/- 15.8 (SE) x 10(-6)] and decreased to lower but still elevated levels at 40 days postpartum. In spleens of mice treated transplacentally with ENU in a split dose, Mfs reached a maximum at 13 days postpartum [74.0 +/- 16.3 (SE) x 10(-6)] and decreased to background levels at 40 days postpartum. The areas under the curves describing the change in hprt Mfs over time for ENU-treated vs. control mice estimate the mutagenic potency for transplacental single- and split-dose exposures to be 1.9 and 0.8 x 10(3), respectively. Comparison of the mutagenic potency estimates for mice exposed to ENU in utero to 4-week-old mice given a similar dose of the same lot number of ENU indicates that the mouse is more susceptible to ENU-induced mutagenesis during fetal life. PMID- 11473386 TI - Micronuclei and chromatid buds are the result of related genotoxic events. AB - Chromatin buds (CHB), broken eggs, or budding cell nuclei are structures similar to micronuclei (MN) in shape, structure, and size, which are linked to the main nuclei of cells by a thread or stalks of chromatin. They have been observed in numerous cell types and there are reports of their existence relating them with MN or with genotoxic events. However, there is no systematic study reporting their frequency and no experiment has been done to ascertain whether they are really induced by genotoxins. Furthermore, they have been discarded as genotoxic events with the argument that they are not formed in dividing cells. Studies are presented here that indicate that CHB can be considered as genotoxic events and that their origin is comparable to that of MN. Bromodeoxyuridine (BrdU) was used to label proliferating lymphocytes, which were later identified by means of an immunohistochemical method, using the H2O2-DAB stain. The results show that CHB are consistently formed where MN are seen. CHB were induced by the clastogen mitomycin C (MMC) as well as by the aneuploidogen colcemid, with frequencies similar to MN in both cases, and to multinucleated cells in the case of colcemid. CHB occur in lymphocytes of smokers with frequencies similar to those of MN, and we found that the infection with Taenia solium metacestodes induced a comparable increase of both MN and CHB frequency in lymphocytes from pigs. PMID- 11473387 TI - Characteristics of mutations generated through digestion with restriction enzyme and ligation in plasmid DNA. AB - Recently, the use of restriction enzymes has been extended to studies in which rare events such as mutation and mistakes in DNA repair are examined. In these studies, the specificity of restriction enzymes becomes critical. To clarify the nature of the rare unexpected events occurring in the process of cutting of DNA with restriction enzymes then ligating it, we studied the molecular characteristics of unexpected plasmid DNAs that were retrieved as mutants of the plasmid after transfection to E. coli. The plasmid used was pUR288, containing lacZ as a marker of mutation. It was digested with restriction enzymes under the conditions recommended by the supplier of the enzymes and under the presence of DMSO, which is known to induce star activity of the enzymes. Comparisons of mutant frequencies and of nucleotide sequences of the mutants found in the different conditions indicated that nonspecific endonucleolytic activity similar to that found under star activity was present under the recommended conditions and, further, was responsible for the creation of deletion-type mutations. The frequency of these events ranged from 10(-5) to 10(-3), depending on the kind of restriction enzymes analyzed. Although the levels of the nonspecificity were not high, they should be considered in assays such as mutation and mistakes in DNA repair, where rare events are examined. PMID- 11473388 TI - Thermolysin improves mutation analysis in skin epidermis from ultraviolet light irradiated Muta Mouse. PMID- 11473389 TI - Flow cytometric enumeration of micronucleated reticulocytes: high transferability among 14 laboratories. AB - This laboratory previously described a single-laser flow cytometric method, which effectively resolves micronucleated erythrocyte populations in rodent peripheral blood samples. Even so, the rarity and variable size of micronuclei make it difficult to configure instrument settings consistently and define analysis regions rationally to enumerate the cell populations of interest. Murine erythrocytes from animals infected with the malaria parasite Plasmodium berghei contain a high prevalence of erythrocytes with a uniform DNA content. This biological model for micronucleated erythrocytes offers a means by which the micronucleus analysis regions can be rationally defined, and a means for controlling interexperimental variation. The experiments described herein were performed to extend these studies by testing whether malaria-infected erythrocytes could also be used to enhance the transferability of the method, as well as control intra- and interlaboratory variation. For these studies, blood samples from mice infected with malaria, or treated with vehicle or the clastogen methyl methanesulfonate, were fixed and shipped to collaborating laboratories for analysis. After configuring instrumentation parameters and guiding the position of analysis regions with the malaria-infected blood samples, micronucleated reticulocyte frequencies were measured (20,000 reticulocytes per sample). To evaluate both intra- and interlaboratory variation, five replicates were analyzed per day, and these analyses were repeated on up to five separate days. The data of 14 laboratories presented herein indicate that transferability of this flow cytometric technique is high when instrumentation is guided by the biological standard Plasmodium berghei. PMID- 11473390 TI - Genetic toxicology testing of the antimalarial drugs chloroquine and a new analog, AQ-13. AB - AQ-13 ([N1-(7-chloro-quinolin-4yl)-3-(N3,N3-diethylamino)propylamine] dihydrochloride trihydrate) is an aminoquinoline antimalarial drug that is effective against chloroquine-resistant strains of Plasmodium falciparum. It is structurally similar to the widely used chloroquine diphosphate (CQ). We evaluated these drugs in the three assays currently recommended by the International Conference on Harmonization (ICH): bacterial mutagenesis in Salmonella typhimurium and Escherichia coli, mammalian cell mutagenesis in L5178Y mouse lymphoma cells, and micronucleus induction in rat bone marrow. A small but statistically significant increase in revertant colonies was produced by CQ with Salmonella tester strain TA98 without metabolic activation (MA) and by AQ-13 with strain TA1537 both with and without MA. In L5178Y cells, testing of CQ and AQ-13 up to cytotoxic concentrations with and without MA produced no increase in mutant colonies and no increase in the numbers of small colonies. Slight decreases in the ratio of polychromatic erythrocytes (PCE) to red blood cells (RBC) were observed in male and female rats treated with CQ and in females only treated with AQ-13; however, none of these changes was statistically significant. No increases in the frequency of micronucleated PCE were observed at any dose level of CQ or AQ-13. Although both CQ and AQ-13 showed weak bacterial mutagenicity, this mutagenic effect was not confirmed in either the mouse lymphoma mutagenesis assay or the micronucleus assay. These results indicate that CQ and AQ-13 should pose minimal risk of genotoxic damage in human populations being administered these drugs. PMID- 11473391 TI - Molecular analysis at the hisD3052 allele of S. typhimurium of mutations induced by aromatic amines, activated by mixed-function oxidases from plants. PMID- 11473392 TI - Glutathione S-transferase T1-null genotype interacts synergistically with heavy smoking on lung cancer risk. AB - We studied the influence of genotype for glutathione S-transferase T1 (GSTT1) on susceptibility to lung cancer among 184 Swedish lung cancer patients (88 never smokers and 96 ever-smokers) and 162 matched population controls (79 never smokers and 83 ever-smokers), with special emphasis on gene-environment interactions. Cases had significantly lower frequency of the GSTT1-null genotype than that of controls among never-smokers (4.6 vs. 16.5%, P = 0.02), whereas the frequencies were very close to each other among smokers (7.4 vs. 7.2%). Cases with high packyears of smoking, however, had a significantly higher frequency of the GSTT1-null genotype compared to that of cases with low packyears (18.3 vs. 5.6%, P = 0.005). Adjusted for age and gender, the GSTT1-null genotype appeared to be protective against lung cancer among never-smokers (odds ratio [OR] = 0.2, 95% confidence interval [CI] = 0.07-0.7), although it was associated with an increased risk for lung cancer among smokers (OR = 2.1, 95% CI = 0.8-5.9), mainly attributed to the group of heavy smokers (>23 packyears; OR = 3.5, 95% CI = 0.7 17.3). Heavy smoking conferred a threefold increased risk for lung cancer (OR = 2.6, 95% CI = 1.3-5.0) among GSTT1-positive individuals, but a ninefold increased risk when combined with the GSTT1-null genotype (OR = 9.3, 95% CI = 1.9-46.3, relative to GSTT1-positive light smokers). This joint effect was further demonstrated by a positive interaction between the GSTT1-null genotype and packyears of smoking. The risk of lung cancer increased steeply with increasing packyears among GSTT1-null smokers, whereas no such effect was seen among GSTT1 positive smokers. We conclude that the GSTT1-null genotype may strengthen the effect of heavy smoking on lung cancer risk. PMID- 11473393 TI - Thermochemical aspects of proton transfer in the gas phase. AB - The beginning of the twentieth century saw the development of new theories of acidity and basicity, which are currently well accepted. The thermochemistry of proton transfer in the absence of solvent attracted much interest during this period, because of the fundamental importance of the process. Nevertheless, before the 1950s, few data were available, either from lattice energy evaluations or from calculations using the emerging molecular orbital theory. Advances in mass spectrometry during the last 40 years allowed studies of numerous systems with better accuracy. Thousands of accurate gas-phase acidities or basicities are now available, for simple atomic and molecular systems and for large biomolecules. The intrinsic effect of structure on the Bronsted basic or acidic properties of molecules and the influence of solvents have been unravelled. In this tutorial, the basics of the thermodynamic principles involved are given, and the mass spectrometric techniques are briefly reviewed. Advances in the design and measurements of gas-phase superacids and superbases are described. Recent studies concerning biomolecules are also evoked. PMID- 11473394 TI - Reactions of BBr(n)(+) (n = 0--2) at fluorinated and hydrocarbon self-assembled monolayer surfaces: observations of chemical selectivity in ion--surface scattering. AB - Ion-surface reactions involving BBr(n)(+) (n = 0--2) with a fluorinated self assembled monolayer (F-SAM) surface were investigated using a multi-sector scattering mass spectrometer. Collisions of the B(+) ion yield BF(2)(+) at threshold energy with the simpler product ion BF(+)* appearing at higher collision energies and remaining of lower abundance than BF(2)(+) at all energies examined. In addition, the reactively sputtered ion CF(+) accompanies the formation of BF(2)(+) at low collision energies. These results stand in contrast with previous data on the ion-surface reactions of atomic ions with the F-SAM surface in that the threshold and most abundant reaction products in those cases involved the abstraction of a single fluorine atom. Gas-phase enthalpy data are consistent with BF(2)(+) being the thermodynamically favored product. The fact that the abundance of BF(2)(+) is relatively low and relatively insensitive to changes in collision energy suggests that this reaction proceeds through an entropically demanding intermediate at the vacuum--surface interface, one which involves interaction of the B(+) ion simultaneously with two fluorine atoms. By contrast with the reaction of B(+), the odd-electron species BBr(+)* reacts with the F-SAM surface to yield an abundant single-fluorine abstraction product, BBrF(+). Corresponding gas-phase ion--molecule experiments involving B(+) and BBr(+)* with C(6)F(14) also yield the products BF(+)* and BF(2)(+), but only in extremely low abundances and with no preference for double fluorine abstraction. Ion--surface reactions were also investigated for BBr(n)(+) (n = 0-2) with a hydrocarbon self-assembled monolayer (H-SAM) surface. Reaction of the B(+) ion and dissociative reactions of BBr(+)* result in the formation of BH(2)(+), while the thermodynamically less favorable product BH(+)* is not observed. Collisions of BBr(2)(+) with the H-SAM surface yield the dissociative ion-surface reaction products, BBrH(+) and BBrCH(3)(+). Substitution of bromine atoms on the projectile by hydrogen or alkyl radicals suggests that Br atoms may be transferred to the surface in a Br-for-H or Br-for-CH(3) transfer reaction in an analogous fashion to known transhalogenation reactions at the F-SAM surface. The results for the H-SAM surface stand in contrast to those for the F-SAM surface in that enhanced neutralization of the primary ions gives secondary ion signals one to two orders of magnitude smaller than those obtained when using the F-SAM surface, consistent with the relative ionization energies of the two materials. PMID- 11473395 TI - Use of L-[(15)N] glutamic acid and homoglutathione to determine both glutathione synthesis and concentration by gas chromatography-mass spectrometry (GCMS). AB - A method for simultaneous measurement of both glutathione enrichment and concentration in a biological sample using gas chromatography mass spectrometry is described. The method is based on the preparation of N,S-ethoxycarbonylmethyl ester derivatives of glutathione, and the use of homoglutathione (glutamyl cysteinyl--alanine) as an internal standard. A procedure for determination of glutamate concentration and enrichment is also reported. Both methods have within day and day-to-day inter-assay coefficients of variation less than 5%, and recoveries of known added amounts of glutathione and glutamate are close to 100%. Taken together, these methods allowed determination of glutathione concentration and fractional synthesis rate in red blood cells using L-[(15)N] glutamic acid infusion. This approach was applied in vivo to investigate the effects of a 72 h fast, compared with a control overnight fast, on erythrocyte glutathione in a single dog. The 72 h fast was associated with a 39% decline in erythrocyte glutathione level, (2.9 +/- 0.4 versus 4.7 +/- 0.5 mmol l(-1), fasting versus control) with no change in glutathione fractional synthesis (67.4 versus 71.3% d( 1), fasting versus control). PMID- 11473396 TI - Determination of famotidine in low-volume human plasma by normal-phase liquid chromatography/tandem mass spectrometry. AB - A rapid, sensitive and robust assay procedure using liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) for the determination of famotidine in human plasma and urine is described. Famotidine and the internal standard were isolated from plasma samples by cation-exchange solid-phase extraction with benzenesulfonic acid (SCX) cartridges. The urine assay used direct injection of a diluted urine sample. The chromatographic separation was accomplished by using a BDS Hypersil silica column with a mobile phase of acetonitrile-water containing trifluoroacetic acid. The MS/MS detection of the analytes was set in the positive ionization mode using electrospray ionization for sample introduction. The analyte and internal standard precursor-product ion combinations were monitored in the multiple-reaction monitoring mode. Assay calibration curves were linear in the concentration range 0.5--500 ng ml(-1) and 0.05--50 microg ml(-1) in plasma and urine, respectively. For the plasma assay, a 100 microl sample aliquot was subjected to extraction. To perform the urine assay, a 50 microl sample aliquot was used. The intra-day relative standard deviations at all concentration levels were <10%. The inter-day consistency was assessed by running quality control samples during each daily run. The limit of quantification was 0.5 ng ml(-1) in plasma and 0.05 microg ml(-1) in urine. The methods were utilized to support clinical pharmacokinetic studies in infants aged 0-12 months. PMID- 11473397 TI - Reaction monitoring of platinum(II) complex--5'-guanosine monophosphate adduct formation by ion exchange liquid chromatography/electrospray ionization mass spectrometry. AB - Cisplatin and four structurally related platinum(II) complexes were incubated with guanosine 5'-monophosphate (5'-GMP) in water at 37 degrees C. The adduct formation reactions were monitored with cation- and anion-exchange liquid chromatography/electrospray ionization mass spectrometry. In addition to mono- and bis-adducts of guanosine 5'-monophosphate with the platinum(II) complexes, other molecular species, presumably with a binuclear structure (two platinum(II) centres), were detected in the reaction mixtures, which have not been reported previously, indicating an unexpected complexity of adduct formation. Anion exchange chromatography revealed the presence of isomers of two complexes which presumably result from the restricted rotation at the platinum-- N-7 (5'-GMP) bonds. All reaction products were characterized in both the positive and negative ion modes. Furthermore, preliminary kinetics and half-times of complex formation were investigated for cisplatin and two other platinum(II) complexes, monitoring the relative concentrations of free 5'-GMP and of mono- and bis-GMP adducts as a function of time (250 h) using an internal standard protocol with thymidine 5' monophosphate. PMID- 11473398 TI - Electron ionization mass spectra of 3,4-disubstituted-1,2,4-oxa(thia)diazole 5(4H)-thione(ones). Substituent effects on the mass spectrometric rearrangement of 3-aryl-4-(p-tolyl)-1,2,4-oxadiazole-5(4H)-thiones to the corresponding oxo compounds. AB - The fragmentation behaviour of ten 3,4-disubstituted 1,2,4-oxadiazole-5(4H) thiones and seven 3,4-disubstituted 1,2,4-thiadiazole-5(4H)-ones studied here confirmed the earlier observations about the partial rearrangement of the former after ionization into the latter before further fragmentation. In the case of eight 3-(substituted phenyl)-4-(p-tolyl)-1,2,4-oxadiazole-5(4H)-thiones the fragmentations reflecting the above-mentioned molecular ion rearrangement show a clear correlation on the substituent sigma values. The electron-withdrawing substituents destabilize the molecular ion, so higher amounts of the rearranged ion [R(1)NCO](+.) are obtained. A good correlation of log[R(1)NCO](+) against sigma was obtained (r = 0.96). Only a satisfactory correlation prevailed for log([R(1)NCO](+)*/[R(1)NCS](+)*) against sigma(r = 0.87). PMID- 11473399 TI - Determination of molecular mass distribution of silicone oils by supercritical fluid chromatography, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and their off-line combination. AB - Silicone oil samples were characterized by supercritical fluid chromatography (SFC), matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI--TOF MS), and their off-line combination. SFC was used to separate samples of silicone oils on micropacked capillary columns. The fractions for the identification studies were obtained from SFC runs at defined time intervals, when the restrictor was pulled out from the chromatographic flame ionization detector (FID) and inserted into a glass vial with acetone. MALDI--TOF MS was used for the identification of individual oligomers in the fractions separated. The molecular mass distributions determined based on SFC and MALDI- TOF MS measurements were compared. From this comparison, it follows that the results are in good agreement. However, certain differences were observed: MALDI- TOF MS was capable of detecting somewhat larger oligomers than the SFC-FID, but the lower molecular mass oligomers were not present in the MALDI spectra. Differences in the region of lower molecular masses can be explained by evaporation of the more volatile low molecular mass oligomers resulting from heating of the sample during the MALDI--TOF MS measurements as a result of the absorption of the laser shot energy. The fact that no high mass discrimination effects of the MALDI--TOF MS measurements, compared with SFC, were observed is very promising for further applications of MALDI--TOF MS in characterizing synthetic polymers of moderate polydispersity. PMID- 11473400 TI - Simultaneous determination of etoposide and its catechol metabolite in the plasma of pediatric patients by liquid chromatography/tandem mass spectrometry. AB - The anticancer drug etoposide is associated with leukemias with MLL gene translocations and other translocations as a treatment complication. The genotype of cytochrome P450 3A4 (CYP3A4), which converts etoposide to its catechol metabolite, influences the risk. In order to perform pharmacokinetic studies aimed at further elucidation of the translocation mechanism, we have developed and validated a liquid chromatography/electrospray/tandem mass spectrometry assay for the simultaneous analysis of etoposide and its catechol metabolite in human plasma. The etoposide analog teniposide was used as the internal standard. Liquid chromatography was performed on a YMC ODS-AQ column. Simultaneous determination of etoposide and its catechol metabolite was achieved using a small volume of plasma, so that the method is suitable for pediatric patients. The limits of detection were 200 ng ml(-1) etoposide and 10 ng ml(-1) catechol metabolite in human plasma and 25 ng ml(-1) etoposide and 2.5 ng ml(-1) catechol metabolite in protein-free plasma, respectively. Acceptable precision and accuracy were obtained for concentrations in the calibration curve ranges 0.2--100 microg ml( 1) etoposide and 10--5000 ng ml(-1) catechol metabolite in human plasma. Acceptable precision and accuracy for protein-free human plasma in the range 25- 15 000 ng ml(-1) etoposide and 2.5--1500 ng ml(-1) etoposide catechol were also achieved. This method was selective and sensitive enough for the simultaneous quantitation of etoposide and its catechol as a total and protein-free fraction in small plasma volumes from pediatric cancer patients receiving etoposide chemotherapy. A pharmacokinetic model has been developed for future studies in large populations. PMID- 11473401 TI - Quadrupole time-of-flight versus triple-quadrupole mass spectrometry for the determination of phosphopeptides by precursor ion scanning. AB - An API 3000 triple-quadrupole instrument and a QSTAR Pulsar quadrupole time-of flight (TOF) mass spectrometer were compared for the determination of phosphopeptides by precursor ion scanning in both the positive and negative nanoelectrospray ionization modes. The limits of detection for synthetic phosphopeptides were similar (500 amol microl(-1)) for both types of instruments when monitoring precursors of -79 Da (PO(3)(-)). However, the quadrupole TOF system was approximately fivefold more sensitive (1 fmol microl(-1)) than the triple-quadrupole instrument (5 fmol microl(-1)) when monitoring precursors of 216 Da (immonium ion of phosphotyrosine). The recently introduced Q(2)-pulsing function, which enhances the transmission of fragment ions of a selected m/z window from the collision cell into the TOF part, improved the sensitivity of precursor ion scans on a quadrupole TOF instrument. The selectivity of precursor ion scans is much better on quadrupole TOF systems than on triple quadrupoles because the high resolving power of the reflectron-TOF mass analyzer permits high accuracy fragment ion selection at no expense of sensitivity. This minimizes interferences from other peptide fragment ions (a-, b-, and y- type) of the same nominal mass but with sufficient differences in their exact masses. As a result, the characteristic immonium ion of phosphotyrosine at m/z 216.043 can be utilized for the selective detection of tyrosine phosphorylated peptides. Our data suggest that, in addition to their superior performance for peptide sequencing, quadrupole TOF instruments also offer a very viable alternative to triple quadrupoles for precursor ion scanning, thus combining high sensitivity and selectivity for both MS and MS/MS experiments in one instrument. PMID- 11473402 TI - Identification of ozone-oxidation products of oxycodone by electrospray ion trap mass spectrometry. AB - The products of oxycodone oxidized by ozone were characterized by electrospray ionization-tandem mass spectrometry (ESI--MS/MS). Liquid Chromatography(LC)--MS analyses revealed that the main constituents in the oxidation reaction mixture included the protonated molecules m/z 316, corresponding to oxycodone, and m/z 332, m/z 348, m/z 366, corresponding to the oxidation products. ESI--MS/MS and MS(n) spectra were used to study oxycodone fragmentation in detail and to characterize the structures of oxidation products. The results show that the oxidation products were formed by addition of one or two oxygen atoms or by addition of three oxygen and two hydrogen atoms to oxycodone. The fragmentation of the oxidation products also shows that the aromatic ring oxidizes due to rupture of the C-3--C-4 bond during product formation. PMID- 11473403 TI - Acetate, propionate and butyrate in plasma: determination of the concentration and isotopic enrichment by gas chromatography/mass spectrometry with positive chemical ionization. AB - This study describes a rapid and simple method to determine short-chain fatty acid (SCFA) concentrations and their isotopic enrichments (M(0) + 1 and M(0) + 2) in human plasma. Sample preparation involves SCFA extraction and derivatization with 1-(tert-butyldimethylsilyl)imidazole. Gas chromatography/mass spectrometry was performed using chemical ionization with ammonia as the reagent gas. Outstanding resolution, excellent linearity and good detection limits were obtained. Inter-assay and intra-assay repeatability was below 10% and 3% respectively for SCFA concentration. Inter-assay repeatability was below 5%, 4%, 6%, and 14% for isotopic enrichment determination of [1-(13)C]acetate and [1,2 (13)C(2)]acetate, [1-(13)C]propionate and [1-(13)C]butyrate respectively, with intra-assay being below 6%. Such SCFA concentrations and isotopic enrichments were determined in the plasma of rats infused with a (13)C-labeled SCFA. The turnovers of acetate, propionate and butyrate in rats were 19 micromol kg(-1) min(-1), 2.6 micromol kg(-1) min(-1), 0.3 micromol kg(-1) min(-1) respectively. PMID- 11473404 TI - Electrospray ionization mass spectrometry of tribenzyltin substituted phenoxyacetate compounds. AB - Hydrolysis products of organotin compounds RC(6)H(4)OCH(2)COOSn(CH(2)ph)(3) (R = o-NO(2), 1; m-NO(2), 2; p-NO(2), 3; o-CH(3), 4; o-OCH(3), 5; o-Cl, 6; o-Br, 7) and RC(6)H(3)OCH(2)COOSn(CH(2)ph)(3) (R = o,o-2CH(3), 8, o-OCH(3), p-CHO, 9; o,p 2Cl, 10), produced in aqueous acetonitrile solution, have been investigated by electrospray mass spectrometry (MS) and MS(n) techniques. The complexes [Y(2)SnXR'](-), [Y(3)SnXR'](-), [Y(3)SnX(2)R'](-), [Y(2)SnX(3)R'](-), and fragment ions of [Y(3)SnR'](-), plus abundant RC(6)H(4)(or RC(6)H(3))OCH(2)COO(-) and RC(6)H(4)(or RC(6)H(3))O(-) ions are observed in negative mode, whereas the protonated molecular ion [M + H](+), complexes [Y(2)SnXR'](+), [Y(3)SnXR'](+), [Y(2)SnX(2)R'](+), [Y(3)SnX(2)R'](+), [Y(2)SnX(3)R'](+), [Y(3)SnX(3)R'](+), as well as [YSnXR'](+), [M - CH(2)ph](+), XSn(+), (phCH(2))(3)Sn(+), phCH(2)Sn(+) (Y = &bond;CH(2)ph, X = &bond;OOCCH(2)OC(6)H(4)R(or C(6)H(3)R)) are detected in the positive mode. Water adduct ions are seen in both modes. The assignments are facilitated by agreement between observed and calculated isotopic patterns and tandem mass spectrometry studies. PMID- 11473405 TI - Metastable ion study of organosilicon compounds. Part XIII: dimethoxydimethylsilane, (CH(3))(2)Si(OCH(3))(2), and dimethoxymethylsilane, CH(3)SiH(OCH(3))(2). AB - Unimolecular metastable fragmentations of dimethoxydimethylsilane, (CH(3))(2)Si(OCH(3))(2) (MW 120, 1), and dimethoxymethylsilane, CH(3)SiH(OCH(3))(2) (MW 106, 2), upon electron impact ionization have been studied by means of mass-analyzed ion kinetic energy (MIKE) spectrometry and the D-labeling technique in conjunction with thermochemistry. The results have been compared with those of the corresponding carbon analogues, 2,2-dimethoxypropane, (CH(3))(2)C(OCH(3))(2) (MW 104, 3) and 1,1-dimethoxyethane, CH(3)CH(OCH(3))(2) (MW 90, 4). In analogy with the cases of 3 and 4, both molecular ions from 1 and 2 are formed at very low abundance at 70 eV, and begin to decompose by the expulsion of the substituents (H, CH(3) or OCH(3)) on the central silicon atom. These decompositions are followed by the loss of a formaldehyde molecule (CH(2)O), as commonly observed in the mass spectra of methoxysilanes. Further, an ethylene (C(2)H(4)) or a dimethyl ether (CH(3)OCH(3)) molecule loss is observed in the fragmentation of some intermediate ions generated from 1(+)* and 2(+)*, but the mechanisms are different than those in the cases of 3 and 4. Some of these fragmentations are also different than those reported previously. The relative abundance of the ions in many MIKE spectra is explained by the extension of the Stevenson-Audier rule. The reaction, which is in contrast to the rule, however, is rationalized by the energy of the transition state for the reaction, estimated by semi-empirical molecular orbital calculation. The peak at m/z 59 from 2(+)* consists only of CH(3)OSi(+) ion, whereas the peak from 1(+)* consists of two different ions, CH(3)OSi(+) and (CH(3))(2)Si(+)H. The ions CH(3)OSi(+) from 1(+)* and 2(+)* are generated by at least two and three separate routes respectively. PMID- 11473406 TI - Characterization by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry of the major photoproducts of temoporfin (m-THPC) and bacteriochlorin (m-THPBC). AB - The photobleaching of 5,10,15,20-tetrakis(m-hydroxyphenyl)chlorin (temoporfin, m THPC) and 5,10,15,20-tetrakis(m-hydroxyphenyl)bacteriochlorin (bacteriochlorin, m THPBC) was studied in ethanol-water (1 : 99, v/v) and in physiological medium (phosphate-buffered saline, PBS) with or without fetal calf serum (FCS). m-THPC solution was irradiated with the laser radiation of 650 nm, whereas m-THPBC solution underwent two consecutive irradiations at 532 and 650 nm. The photoproducts were characterized by UV-visible absorption spectrophotometry and by matrix-assisted laser desorption/ionization (MALDI) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS). Independent of the solvent used, the phototransformation of either photosensitizer yielded the formation of 5,10,15,20-tetrakis (m-hydroxyphenyl)porphyrin (m-THPP) through a major dehydrogenation process. PMID- 11473407 TI - Labile sulfogroup allows differentiation of sulfotyrosine and phosphotyrosine in peptides. PMID- 11473408 TI - Measurement of chiral complexes of cyclodextrins and amino acids by electrospray ionization time-of-flight mass spectrometry. PMID- 11473409 TI - Current literature in mass spectrometry. PMID- 11473410 TI - New natural epothilones from Sorangium cellulosum, strains So ce90/B2 and So ce90/D13: isolation, structure elucidation, and SAR studies. AB - In addition to epothilones A (1) and B (2), 37 natural epothilone variants and epothilone-related compounds were isolated from the culture broth of a 700 L fermentation of Sorangium cellulosum, strain So ce90/B2. Of these, only the 12,13 desoxyepothilones, epothilone C (14) and D (15), were produced in significant amounts (3-6 mg/L); the 21-hydroxy derivatives and epothilones E (3) and F (4), in low and variable amounts due to further degradation by the producing organism. Most of the other epothilone variants were produced only in 1-100 microg/L amounts. The new compounds are very similar in structure to the parent compounds 1, 2 and 14, 15 and are presumably the result of the imperfect selectivity of the biosynthetic enzymes for acetate and propionate. Further, epothilones containing an oxazole moiety (10-13) in the side chain instead of a thiazole as well as ring expanded 18-membered macrolides, epothilones I (30-35), and a ring contracted 14 membered macrolide, epothilone K (36), were found as very minor metabolites. The mutant strain, So ce90/D13, instead of macrolactones, produced short-chain carboxylic acids 40, 41, and 42 bearing the characteristic thiazole side chain. The structures of the new epothilones were elucidated on the basis of comprehensive NMR and MS data. The new epothilone variants were tested in a cytotoxicity assay with mouse fibroblasts (cell line L929), and structure activity relationships were established. Several new natural epothilones showed activity comparable to 1 and 2, but in no case exceeded that of 2. PMID- 11473411 TI - Four new flavones and a new isoflavone from Iris bungei. AB - Four new irisflavones A-D (1-4) and irilin D (5) have been isolated from the underground parts of Iris bungei along with known isoflavones, irilins A-B (6-7) and tlatancuayin (8). The structures of the new compounds were determined using NMR and mass spectroscopic methods and were found to be 2',5,7-trihydroxy-3,6 dimethoxyflavone (1), 2',5-dihydroxy-3,6,7-trimethoxyflavone (2), 2',5,6' trihydroxy-3,6,7-trimethoxyflavone (3), 3,3',5-trihydroxy-2',7-dimethoxyflavone (4), and 3',4',5,7-tetrahydroxy-6-methoxyisoflavone (5). The structures of irisflavones 1, 3, and 4 were confirmed by single-crystal X-ray diffraction studies. PMID- 11473412 TI - Antiinflammatory constituents from Heterotheca inuloides. AB - Three new compounds, cadalen-15-oic acid (1), 3,7-dihydroxy-3(4H)-isocadalen-4 one (2), and dicadalenol (3), were isolated from the aerial parts of Heterotheca inuloides (Mexican arnica), together with the known compounds 7-hydroxycadalene (4), 7-hydroxy-4alphaH-3,4-dihydrocadalene (5), 1alpha-hydroxy-1(4H)-isocadalen-4 one (6), 1alpha-hydroxy-4alphaH-1,2,3,4-tetrahydrocadalen-15-oic acid (7), 7-(3,3 dimethylallyloxy)coumarin, caryolan-1,9beta-diol, and quercetin. The structures of the new compounds were elucidated by spectroscopic methods. The antiinflammatory activities of the extracts and the isolated compounds were evaluated by determining the inhibition of TPA-induced mouse ear edema. The natural products 3, caryolan-1,9beta-diol, and quercetin were the most active substances tested and displayed dose-dependent activities. PMID- 11473413 TI - Monoterpene glycosides and triterpene acids from Eriobotrya deflexa. AB - A phytochemical study on a methanolic extract of leaves of Eriobotrya deflexa led to the isolation and characterization of nine terpenoid compounds. Four of these are new chemical entities, including two monoterpene glycosides, (3S)-O-alpha-L rhamnopyranosyl-(1-->3)-[4-O-(E)-coumaroyl]-alpha-L-rhamnopyranosyl-(1-->6)-beta D-glucopyranosyl-linalool (1) and (3S)-O-alpha-L-rhamnopyranosyl-(1-->3)-[4-O-(Z) coumaroyl]-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranosyl-linalool (2), and two triterpene acids, 1beta,2alpha,19alpha-trihydroxy-3-oxo-12-ursen-28-oic acid (3) and 2alpha,3alpha,19alpha-trihydroxy-12-oleanen-28-oic acid (4). Their structures were elucidated on the basis of spectroscopic analysis. The activities of these isolates in an in vitro antiproliferation test were also determined. PMID- 11473414 TI - Novel sesquiterpenoids from the roots of Phyllanthus emblica. AB - Three novel bisabolane-type sesquiterpenoids, phyllaemblic acids B (1) and C (2) and phyllaemblicin D (3), together with two new phenolic glycosides, 2 carboxylmethylphenol 1-O-beta-D-glucopyranoside (4) and 2,6-dimethoxy-4-(2 hydroxyethyl)phenol 1-O-beta-D-glucopyranoside (5), were isolated from the roots of Phyllanthus emblica. The structures of 1-5 were established by spectral and chemical methods. The absolute stereochemistry of 1 and 2 was determined by applying the PGME method. PMID- 11473415 TI - The complestatins as HIV-1 integrase inhibitors. Efficient isolation, structure elucidation, and inhibitory activities of isocomplestatin, chloropeptin I, new complestatins, A and B, and acid-hydrolysis products of chloropeptin I. AB - From the screening of a microbial extract library, isocomplestatin (1), a new axial-chiral isomer of complestatin (2) which is a known rigid bicyclic hexapeptide, was identified as a potent natural product inhibitor of HIV-1 integrase, a unique enzyme responsible for viral replication. Isocomplestatin showed inhibitory activities (IC(50)) in coupled 3'-end processing/strand transfer (200 nM), strand transfer (4 microM), and HIV-1 replication (200 nM) in virus-infected cells. Attempted large-scale isolation of 1 by the literature method, used for the isolation of complestatin, led to lower yield and limited availability. We have developed several new, two-step, high-yielding absorption/elution methods of isolation based on reverse-phase chromatography at pH 8 that are applicable to scales from one gram to potential industrial quantities. We have also discovered and determined the structure of two new congeners of 1, namely, complestatins A (4) and B (5), with almost equal HIV-1 integrase activity. They differ from 1 at C2' and C3' of the tryptophan moiety (residue F). Selective acid hydrolysis of chloropeptin I (3), itself a known acid catalyzed rearranged isomer of 1 and 2 (8'- vs 7'-substitution in tryptophan residue F, respectively), an isomer of complestatin, and isocomplestatin resulted in a number of fragments (6-10) with retention of most of the HIV-1 integrase activity. The structure-activity relationship as revealed by these compounds could possibly lead to the design of better inhibitors or understanding of the HIV-1 integrase target. PMID- 11473416 TI - Synthesis of phakellistatin 11: a micronesia (Chuuk) marine sponge cyclooctapeptide. AB - The cyclic octapeptide phakellistatin 11 (1), a constituent of The Federated States of Micronesia (Chuuk) marine sponge Phakellia sp., was synthesized using solid-phase techniques. An initial solution-phase synthesis proved to be inadequate owing to spontaneous deprotection of the Fmoc group at the heptapeptide stage. Using the PAL resin attachment and proceeding from Fmoc-Glu alpha-allyl ester, linear elongation of the octapeptide was performed until the final unit Pro was added. The allyl ester was removed using Pd(0)[P(C(6)H(5))(3)](4). Cleavage of the final Fmoc group and cyclization with PyAOP provided phakellistatin 11 (1) in 17% overall yield. The synthetic specimen of phakellistatin 11 (1) was found to be chemically but not biologically (cancer cell lines) identical to the natural product. The result suggested a conformational difference or more likely the presence of a trace amount of a highly active antineoplastic agent that binds noncovalently to the natural cyclic octapeptide 1. PMID- 11473417 TI - Antioxidant principles from Bauhinia tarapotensis. AB - A new cyclohexenone (1) and a new caffeoyl ester derivative (2), together with the known compounds (-)-isolariciresinol 3-alpha-O-beta-D-glucopyranoside (3), (+)-1-hydroxypinoresinol 1-O-beta-D-glucopyranoside (4), isoacteoside (5), luteolin 4'-O-beta-D-glucopyranoside (6), and indole-3-carboxylic acid (7), were isolated from the leaves of Bauhinia tarapotensis. The structures of these new compounds were determined by spectroscopic data analysis. The antioxidant activities of 1-7 were determined by measuring their free radical scavenging effects, using the 1,1-diphenyl-2-dipicrylhydrazyl free radical (DPPH) and Trolox equivalent antioxidant activity (TEAC) methods, and the coupled oxidation of beta carotene and linoleic acid. Compounds 3-5 showed good activities in the DPPH and TEAC tests, while compounds 1 and 2 were active in the coupled oxidation of beta carotene and linoleic acid bioassay. PMID- 11473418 TI - In vitro cytotoxicity of the protoberberine-type alkaloids. AB - In vitro cytotoxic activities of 24 quaternary protoberberine alkaloids related to berberine have been evaluated using a human cancer cell line panel coupled with a drug sensitivity database. Extending the alkyl chain at position 8 or 13 strongly influenced the cytotoxic activity, that is, relative lipophilicity as well as the size of the substituent affects cytotoxicity. The highest level of activity was observed in 8- or 13-hexyl-substituted derivatives of berberine. Structure-activity relationships are described. PMID- 11473419 TI - Diterpenoids from Acacia leucophloea: revision of the structures of leucophleol and leucophleoxol. AB - The pimarane-type structures previously suggested for leucophleol (1) and leucophleoxol (2), two diterpenoids isolated from Acacia leucophloea, must be amended to the isopimarane-type derivatives 5 and 6, respectively. These corrections were supported on NMR spectroscopic studies and, in the case of 6, by an X-ray diffraction analysis. Moreover, the unpublished complete and unambiguous (1)H and (13)C NMR assignments of 5 and 6 together with those of leucoxol (3), another diterpenoid from the same plant, are also reported. PMID- 11473420 TI - Three xanthones and a benzophenone from Garcinia mangostana. AB - Investigation of the constituents of Garcinia mangostana has led to the isolation of four new compounds: three minor xanthones, garcimangosone A (1), garcimangosone B (2), and garcimangosone C (3), and a benzophenone glucoside, garcimangosone D (4). The structures of these four compounds were established by spectral (NMR and MS) and chemical methods. PMID- 11473421 TI - Isolation of dolastatin 10 from the marine cyanobacterium Symploca species VP642 and total stereochemistry and biological evaluation of its analogue symplostatin 1. AB - The potent antitumor agent dolastatin 10 (1) was originally isolated from the sea hare Dolabella auricularia, and we now report its isolation from the marine cyanobacterium Symploca sp. VP642 from Palau. The chemically related analogue symplostatin 1 (2) has been reisolated from Guamanian and Hawaiian varieties of S. hydnoides and its total stereochemistry completed by determining the N,N dimethylisoleucine unit to be L. Symplostatin 1 (2), like dolastatin 10 (1), is a potent microtubule inhibitor. The antitumor activity of 2 was assessed in vivo against several murine tumors. Symplostatin 1 (2) was effective against a drug insensitive mammary tumor and a drug-insensitive colon tumor; however, it was only slightly effective against two MDR tumors. PMID- 11473422 TI - Biological and mechanistic activities of xanthorrizol and 4-(1',5'-dimethylhex-4' enyl)-2-methylphenol isolated from Iostephane heterophylla. AB - Xanthorrizol (1) and 4-(1',5'-dimethylhex-4'-enyl)-2-methylphenol (2) were identified as the principal antimicrobial components of a CH(2)Cl(2)-MeOH (1:1) extract derived from Iostephane heterophylla. Compound 2 is a new natural product, but has been synthesized. Both compounds exhibited low level activity (MICs of 16-32 microg/mL) against methicillin-resistant staphylococci and vancomycin-resistant enterococci. They were either inactive or poorly active against Gram-negative bacteria and yeast. Mechanistic studies performed in Escherichia coli imp suggested nonspecific inhibition of DNA, RNA, and protein synthesis by both of these compounds. Compound 1 was tested in an in vivo model; it did not provide protection to mice infected with Staphylococcus aureus. PMID- 11473423 TI - Cytotoxic pheophorbide-related compounds from Clerodendrum calamitosum and C. cyrtophyllum. AB - Three pheophorbide-related compounds (1-3) were isolated from the leaves and stems of Clerodendrum calamitosum. The methyl ester of 3 (6) and the known (10S) hydroxypheophytin a (7) also were isolated from leaves of the related plant Clerodendrum cyrtophyllum. Compounds 1 and 6 were isolated for the first time as naturally occurring products from a plant source. All structures were elucidated by detailed spectroscopic analysis. Biological evaluation showed that 1 and 2 exhibited strong cytotoxicity against human lung carcinoma (A549), ileocecal carcinoma (HCT-8), kidney carcinoma (CAKI-1), breast adenocarcinoma (MCF-7), malignant melanoma (SK-MEL-2), ovarian carcinoma (1A9), and epidermoid carcinoma of the nasopharynx (KB), and its etoposide- (KB-7d), vincristine- (KB-VCR), and camptothecin-resistant (KB-CPT) subclones. Compound 3 was less cytotoxic than 1 and 2. Compounds 4-6, the methyl esters of 1-3, showed strongly increased cytotoxicity compared with the parent acids. Interestingly, 6 was the most active derivative among these compounds. Compound 7 was inactive. PMID- 11473424 TI - Three new acylated triterpene saponins from Acanthophyllum squarrosum. AB - Three new triterpenoid saponins, 1-3, were isolated from the roots of Acanthophyllum squarrosum. Their structures were established mainly by 2D NMR techniques as 3-O-beta-D-galactopyranosyl-(1-->2)-[beta-D-xylopyranosyl-(1-->3)] beta-D-glucuronopyranosyl-gypsogenin-28-O-beta-D-xylopyranosyl-(1-->3)-beta-D xylopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)-3-O-acetyl-alpha-L rhamnopyranosyl-(1-->2)-3,4-di-O-acetyl-beta-D-fucopyranoside (1), 3-O-beta-D galactopyranosyl-(1-->2)-[beta-D-xylopyranosyl-(1-->3)]-beta-D-glucuronopyranosyl gypsogenin-28-O-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->2)-[5-O acetyl-alpha-L-arabinofuranosyl-(1-->3)]-4-O-acetyl-beta-D-fucopyranoside (2), and 3-O-beta-D-glucopyranosyl-quillaic acid-28-O-alpha-L-rhamnopyranosyl-(1-->2) alpha-L-arabinopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->6)]-beta-D glucopyranoside (3). PMID- 11473425 TI - Novel cytotoxic annonaceous acetogenins from Annona muricata. AB - Seven new annonaceous acetogenins, muricins A-G (1-7), as well as five known compounds, a mixture of muricatetrocin A (8) and muricatetrocin B (9), longifolicin (10), corossolin (11), and corossolone (12), were isolated from the seeds of Annona muricata. The structures of all isolates were elucidated and characterized by spectral and chemical methods. These acetogenins showed significantly selective in vitro cytotoxicities toward the human hepatoma cell lines Hep G(2) and 2,2,15. PMID- 11473426 TI - Antimicrobial cuparene-type sesquiterpenes, enokipodins C and D, from a mycelial culture of Flammulina velutipes. AB - Two new cuparene-type sesquiterpenes, enokipodins C (1) and D (2), were isolated from culture medium of an edible mushroom, Flammulina velutipes, along with enokipodins A (3) and B (4). The structures of 1 and 2 were determined using spectroscopic methods (HRMS, (1)H and (13)C, and 2D NMR). The absolute configuration of enokipodin C was determined from the observed (1)H NMR chemical shifts and NOEs in NOESY experiments after conversion into the corresponding esters with the chiral reagent 2-(2'-methoxy-1'-naphthyl)-3,4-dichlorobenzoic acid. All the metabolites showed antimicrobial activity against a fungus, Cladosporium herbarum, and Gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus. PMID- 11473427 TI - Seven new sesquiterpene glycosides from the root bark of Dictamnus dasycarpus. AB - From the water-soluble constituents of the root bark of Dictamnus dasycarpus, six new eudesmane-type sesquiterpene glycosides, dictamnosides H-M (1-6), and a new trinorguaiane-type sesquiterpene glycoside, dictamnoside N (7), together with four known sesquiterpene glycosides, dictamnosides A (8), B (9), D (10), and G (11), were isolated. Their structures were elucidated by spectroscopic analyses and chemical evidence. In vitro tests for immunological activity showed dictamnoside A (8) to possess remarkable activity in stimulating the proliferation of T-cells. PMID- 11473428 TI - New carbonimidic dichlorides from the Australian sponge Ulosa spongia and their possible taxonomic significance. AB - Five sesquiterpene carbonimide dichlorides (1-5) have been isolated from the dichloromethane extract of the Australian sponge Ulosa spongia. The structures of the two new compounds, ulosins A (1) and B (2), were elucidated by employing spectroscopic techniques (NMR, MS, UV, and IR). The chemotaxonomic significance of our data is discussed. PMID- 11473429 TI - Prunioside A: a new terpene glycoside from Spiraea prunifolia. AB - Prunioside A (1) has been isolated from an EtOAc-soluble extract of Spiraea prunifolia var. simpliciflora by a combination of chromatographic techniques. The structure was determined primarily by extensive NMR experiments. Compound 1 is a unique terpene glycoside. Its acetylated derivative (1a) inhibited nitric oxide production in murine macrophage-like RAW 264.7 cells in a dose-dependent manner. PMID- 11473430 TI - New steroid glycosides from the starfish Asterias rathbuni. AB - Two novel steroidal 24-O-xylosides, designated as rathbuniosides R(1) (1) and R(2) (2), and the known amurensoside A (3) and 3-O-sulfomarthasterone (4) have been isolated from the starfish Asterias rathbuni. The structures of all the compounds were determined from their spectroscopic data, including one- and two dimensional NMR methods. The compounds 1 and 4 inhibit the cell division of fertilized sea urchin eggs at doses of 7.0 x 10(-5) and 2.9 x 10(-5) M, respectively. PMID- 11473431 TI - Hydrachine A, a novel alkaloid from the roots of Hydrangea chinensis. AB - A novel alkaloid, hydrachine A (3), has been isolated, along with 15 known compounds, from the roots of Hydrangea chinensis. The structure and stereochemistry of the new alkaloid 3 was determined using extensive 2D NMR data. PMID- 11473432 TI - Taxane diterpenoids from the stem bark of Taxus mairei. AB - Three new 11(15-->1)-abeo-taxanes, taxumairols U-W (1-3), have been isolated from extracts of the stem bark of Formosan Taxus mairei. The structures of 1-3 were identified as 5alpha,7beta,9alpha,13alpha,20-pentaacetoxy-2alpha,10beta,15 trihydroxy-11(15-->1)-abeo-taxene, 5alpha,7beta,9alpha,20-tetraacetoxy 2alpha,10beta,13alpha,15-tetrahydroxy-11(15-->1)-abeo-taxene, and 2alpha,4alpha,7beta,10beta-tetraacetoxy-5beta,20-epoxy-9alpha,13alpha,15 trihydroxy-11(15-->1)-abeo-taxene, respectively, on the basis of 2D NMR techniques including COSY, HSQC, HMBC, and NOESY experiments as well as chemical reactions of compounds 1-3 to give 4 (5alpha,7beta,9alpha,10beta,13alpha,20 hexaacetoxy-2alpha,15-dihydroxy-11(15-->1)-abeo-taxene) and 5 (4alpha,7beta,10beta-triacetoxy-9alpha,13alpha-dibenzoxy-5beta,20-epoxy-2alpha,15 dihydroxy-11(15-->1)-abeo-taxene), which are also novel taxane derivatives. Taxumairols U (1) and V (2) exhibited significant cytotoxicities against human hepatoma tumor cells, while taxumairol W (3) was inactive. PMID- 11473433 TI - Triterpenoids from Tillandsia fasciculata. AB - The leaves of Tillandsia fasciculata afforded four tetracyclic triterpenoids of the cycloartane type, two new compounds identified as cyclolaudenyl formate (2) and the (24S)-24-isopropenyl cycloartanone, which we named tillandsinone (1), and the known cyclolaudenone (3) and cyclolaudenol (4). PMID- 11473434 TI - Montipyridine, a new pyridinium alkaloid from the stony coral montipora species. AB - A new pyridinium alkaloid, montipyridine (1), has been isolated from the stony coral Montipora sp. The structure was established from spectroscopic data. PMID- 11473435 TI - Anti-AIDS agents. 46. Anti-HIV activity of harman, an anti-HIV principle from Symplocos setchuensis, and its derivatives. AB - Matairesinol (1) and harman (5), identified from Symplocos setchuensis, were found to inhibit HIV replication in H9 lymphocyte cells. Anti-HIV evaluation of 28 derivatives of 5 revealed that compound 19 showed potent activity with EC(50) and therapeutic index values of 0.037 microM and 210, respectively. PMID- 11473436 TI - Flavonoid, iridoid, and lignan glycosides from Putoria calabrica. AB - From the aerial parts of Putoria calabrica, two new flavonol triglycosides were isolated and their structures were elucidated as quercetin-3-O-[alpha-L rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside]-7-O-beta-D-glucopyranoside (1, calabricoside A) and quercetin-3-O-[4' "-O-caffeoyl-alpha-L-rhamnopyranosyl-(1- >2)-alpha-L-arabinopyranoside]-7-O-beta-D-glucopyranoside (2, calabricoside B). Additionally, seven iridoid and three lignan glycosides were isolated and characterized. Radical scavenging activities of all compounds were determined by quantifying their effects on luminol-enhanced chemiluminescence in formyl methionyl-leucyl-phenylalanine (FMLP) stimulated human polymorphonuclear neutrophils (PMNs). Calabricoside A and B showed strong radical scavenging activity with IC(50) values of 0.25 and 0.3 microM, respectively. PMID- 11473437 TI - Multiplolides A and B, new antifungal 10-membered lactones from Xylaria multiplex. AB - Two new 10-membered lactones, namely, multiplolides A (1) and B (2), were isolated from the broth extract of the fungus Xylaria multiplex BCC 1111. Chemical structures of 1 and 2 were elucidated on the basis of their spectral data. Multiplolides A (1) and B (2) exhibited antifungal activity against Candida albicans with IC(50) values of 7 and 2 microg/mL, respectively. Both 1 and 2 were inactive in the screening systems toward the malarial parasite Plasmodium falciparum (at 20 microg/mL) and were not cytotoxic to BC-1 and KB cell lines (at 20 microg/mL). PMID- 11473438 TI - Cytotoxic oxoisoaporphine alkaloids from Menispermum dauricum. AB - Four new oxoisoaporphine alkaloids, daurioxoisoporphines A-D (1-4), were isolated from the rhizomes of Menispermum dauricum. The structures of these alkaloids were established by spectroscopic methods. The cytotoxic evaluation of 1 and 2 is reported against four cancer cell lines. PMID- 11473439 TI - A novel dimeric diterpene from Salvia prionitis. AB - A novel dimeric abietane diterpene, hongencaotone (1), has been isolated from the roots of Salvia prionitis, and its structure was determined by spectral data interpretation and X-ray analysis. Hongencaotone (1) is the first ether-linked heterodimeric diterpene to have been isolated from the genus Salvia. PMID- 11473440 TI - Aristophenones A and B. A new tautomeric pair of polyisoprenylated benzophenones from Garcinia aristata. AB - A tautomeric pair of new isoprenylated benzophenones, aristophenone A (1a) and B (1b), have been isolated from Garcinia aristata fruits. Their structures has been determined using high-field 2D NMR techniques. PMID- 11473441 TI - Isolation and bioactivities of constitutents of the roots of Garcinia atroviridis. AB - Two new prenylated compounds, the benzoquinone atrovirinone (1) and the depsidone atrovirisidone (2), were isolated from the roots of Garcinia atroviridis. Their structures were determined on the basis of the analysis of spectroscopic data. While compound 2 showed some cytotoxicity against HeLa cells, both compounds 1 and 2 were only mildly inhibitory toward Bacillus cereus and Staphylococcus aureus. PMID- 11473442 TI - Suberedamines A and B, new bromotyrosine alkaloids from a sponge Suberea species. AB - Two new cytotoxic bromotyrosine alkaloids, suberedamines A (1) and B (2), have been isolated from an Okinawan marine sponge Suberea sp. The structures were elucidated on the basis of spectroscopic data and chemical means. PMID- 11473443 TI - Antillatoxin B, a neurotoxic lipopeptide from the marine cyanobacterium Lyngbya majuscula. AB - Bioassay-guided fractionation of organic extracts from two Lyngbya majuscula collections led to the isolation of a new secondary metabolite, antillatoxin B, an unusual N-methyl homophenylalanine analogue of the potent neurotoxin antillatoxin. Its structure was deduced from 2D NMR and data comparisons with antillatoxin. Antillatoxin B exhibited significant sodium channel-activating (EC(50) = 1.77 microM) and ichthyotoxic (LC(50) = 1 microM) properties. PMID- 11473444 TI - Actaeaepoxide 3-O-beta-D-xylopyranoside, a new cycloartane glycoside from the rhizomes of Actaea racemosa (Cimicifuga racemosa). AB - A new cycloartane glycoside (1) was obtained from a minor triterpene fraction of the rhizome extract of Actaea racemosa (synonym: Cimicifuga racemosa) along with a known compound, cimigenol 3-O-beta-D-xylopyranoside. The structure of 1 was elucidated as 20(S),22(R),23(R),24(S)-12beta-acetoxy-16beta:23,23alpha:24-diepoxy 3beta,22beta,25-trihydroxy-9,19-cyclolanost-7-ene 3-O-beta-D-xylopyranoside (actaeaepoxide 3-O-beta-D-xylopyranoside) on the basis of spectral and chemical evidence. PMID- 11473445 TI - Water-soluble glycosides from Ruta graveolens. AB - An EtOH extract of the dried aerial parts of Ruta graveolens was suspended in water and then partitioned with EtOAc. Three new glycosides, 3'-sinapoyl-6 feruloylsucrose (4), methylcnidioside A (5), and methylpicraquassioside A (6), together with four known glycosides, 3',6-disinapoylsucrose (1), cnidioside A (2), rutin, and picraquassioside A (3), were isolated from the water-soluble part. Their structures were elucidated by interpretation of IR, MS, and 1D and 2D NMR spectra and comparison with literature data. PMID- 11473446 TI - Suppressors of cancer cell proliferation from fig (Ficus carica) resin: isolation and structure elucidation. AB - A mixture of 6-O-acyl-beta-D-glucosyl-beta-sitosterols, the acyl moeity being primarily palmitoyl and linoleyl with minor amounts of stearyl and oleyl, has been isolated as a potent cytotoxic agent from fig (Ficuscarica) latex and soybeans. Identity was established by spectroscopic methods (NMR, MS) and confirmed by chemical synthesis. Both the natural and the synthetic compounds showed in vitro inhibitory effects on proliferation of various cancer cell lines. PMID- 11473447 TI - Cyclopeptides from the bark of Discaria americana. AB - Two new cyclopeptides, named discarene C (1) and discarene D (2), have been isolated from the bark of Discaria americana, along with seven known cyclopeptide alkaloids. The structures of the new compounds were determined by spectroscopic methods, mainly NMR. The stereochemistry of the ring amino acid residues have been assigned by gas chromatography employing modified cyclodextrins as chiral stationary phases. PMID- 11473448 TI - Evaluation of sentinel lymph nodes in colorectal carcinoma. PMID- 11473450 TI - Granular cell traumatic neuroma of salivary gland. PMID- 11473451 TI - Decreasing incidence of sudden death due to undiagnosed primary central nervous system tumors. AB - CONTEXT: Although most fatal brain tumors are diagnosed well before a patient's death, occasionally medical examiners and coroners encounter cases in which the presence of a primary tumor of the central nervous system (CNS) was not suspected prior to death. Analysis of such cases can shed light on specific pitfalls hindering the diagnosis of brain tumors. In addition, by analyzing the incidence of these cases in a large autopsy series, one can draw conclusions about the evolving effectiveness of medical diagnosis. OBJECTIVE: To determine the incidence of deaths due to undiagnosed primary CNS tumors in the era of advanced neuroimaging techniques. DESIGN: Records from forensic autopsies performed during a 20-year period (1980-1999) at the Office of the Chief Medical Examiner of the State of Maryland were reviewed to identify cases in which death was caused by primary CNS tumors undiagnosed prior to the patient's death. RESULTS: We present 11 cases of undiagnosed primary CNS tumors resulting in sudden death that were identified among 54 873 forensic autopsies. Sudden deaths due to undiagnosed CNS neoplasms account for a significantly lower percentage of cases in our study (0.02%) than in similar series reported prior to 1980 (> or =0.16%). CONCLUSIONS: We hypothesize that improvements in imaging techniques, notably the introduction of computed tomography and magnetic resonance imaging, have resulted in increased early detection of CNS neoplasms. However, vague or short-term symptoms and limited health care access can dissuade patients from seeking medical attention and result in failure to diagnose these tumors correctly. PMID- 11473452 TI - Diagnostic utility of intracytoplasmic lumen and transgressing vessels in evaluation of Hurthle cell lesions by fine-needle aspiration. AB - INTRODUCTION: Recent abstracts have emphasized the importance of recognizing intracytoplasmic lumen and transgressing vessels as useful criteria enabling distinction between Hurthle cells encountered in neoplastic and nonneoplastic thyroid aspirates. The purpose of this retrospective study was to evaluate if application of these criteria improves specificity and sensitivity of cytologic diagnosis of true Hurthle cell neoplasms. MATERIALS AND METHODS: We retrospectively reviewed 30 fine-needle aspirates of thyroid with cytologic diagnosis of Hurthle cell neoplasms (13 cases) and nonneoplastic thyroid with prominent Hurthle cells (17 cases). All cases were evaluated for the presence of intracytoplasmic lumen and transgressing vessels and were reclassified as neoplastic or nonneoplastic based on the presence or absence of 1 or both of these criteria. Surgical follow-up was available in all cases. RESULTS: Surgical follow-up in 13 cases of Hurthle cell neoplasms revealed Hurthle cell carcinoma (3 cases), Hurthle cell adenoma (6 cases), and Hashimoto's thyroiditis (4 cases). Seventeen cases with nonneoplastic diagnosis revealed Hurthle cell carcinoma (1 case), Hashimoto's thyroiditis (12 cases), and nodular goiter (4 cases). After application of the previously mentioned cytologic criteria, the cytologic diagnoses were reclassified as Hurthle cell neoplasms (13 cases) and nonneoplastic thyroid (17 cases). The true sensitivity of the test before and after the application of the criteria was 90% and 100%, respectively. The true specificity before and after the application of the cytologic criteria was 65% and 85%, respectively. CONCLUSIONS: Intracytoplasmic lumen and transgressing vessels are helpful features in distinguishing neoplastic and nonneoplastic Hurthle cell thyroid lesions. Use of these criteria may improve the specificity and sensitivity of the cytologic diagnosis. PMID- 11473453 TI - CD30 expression in follicular lymphoma. AB - CONTEXT: CD30(+) anaplastic large cell lymphomas were originally described as being of T-cell, null cell, and B-cell origin. CD30, however, is not a specific marker of anaplastic large cell lymphoma and has been found to be expressed in reactive as well as neoplastic populations as a probable activation marker. In addition, CD30(+) cells have also been described in both diffuse large B-cell and follicular lymphomas (FLs), resembling the pattern seen in reactive tonsils and lymph nodes. OBJECTIVE: We report an index case of FL with CD30 expression, which on initial touch preparations and flow cytometric immunophenotyping revealed a prominent population of CD30(+) cells with marked cellular pleomorphism (anaplasia) in a background of typical FL. Immunohistochemistry of the paraffin section for CD30 in our index case confirmed unequivocal CD30(+) pleomorphic cells in the malignant nodules in occasional clusters. This case prompted a study of additional cases of FL for pattern of immunoreactivity with CD30 on paraffin sections. DESIGN: Twenty-two additional cases of FL (grades 1-3) were retrieved for CD30 immunoperoxidase staining as in the index case. RESULTS: This study demonstrated 32% of the additional cases of FL had definitive CD30(+), large, pleomorphic malignant cells by paraffin immunohistochemistry. In 2 cases (9%), the pattern of immunoreactivity with CD30 showed clustering and variable staining of large cells, as our index case. CONCLUSION: This study underscores the morphologic and immunophenotypic spectrum of FL that includes CD30 staining and cellular pleomorphism. PMID- 11473454 TI - The histopathology of 103 consecutive colonoscopy biopsies from 82 symptomatic patients with acquired immunodeficiency syndrome: original and look-back diagnoses. AB - OBJECTIVE: To compare the primary diagnoses assigned by general surgical pathologists on a series of 103 consecutive colon biopsies from individuals infected with human immunodeficiency virus (HIV) with diagnoses rendered by a pathologist with extensive experience in gastrointestinal pathology in HIV/acquired immunodeficiency syndrome. DESIGN: New sections were cut from paraffin blocks of 103 consecutive colon biopsies taken during colonoscopies of 82 different HIV-infected patients; all new sections were stained with hematoxylin-eosin. These individuals either had negative stool studies or had failed to respond to therapy and had chronic large bowel symptoms, such as frequent small volume-type diarrhea, tenesmus, and/or bright red blood per rectum. Immunohistochemistry for cytomegalovirus (CMV) was performed on 18 of 22 specimens originally diagnosed with CMV colitis. RESULTS: The initial study yielded 70 (68%) negative or nonspecific diagnoses, 22 (21%) cases of CMV colitis, 5 (5%) Cryptosporidium diagnoses, 2 cases each of adenomatous polyps and Kaposi sarcoma, and 1 case each of spirochetosis and squamous cell carcinoma of the anorectum. Review of the recuts yielded 64 (62%) negative or nonspecific diagnoses, 12 (12%) new adenovirus infections (3 combined with CMV), and 11 (11%) lone CMV infections. Three attaching and effacing bacterial infections were diagnosed, 1 with adenovirus coinfection. A total of 4 spirochetosis cases were found on review. Seven (7%) of the biopsies showed at least 1 coinfection. Nine biopsies had features suggestive of inflammatory bowel disease. CONCLUSIONS: Colonoscopy with biopsy after negative stool studies or failure to respond to therapy yielded a high proportion of negative or nonspecific diagnoses. Adenovirus and enteropathogenic bacterial infections had been totally overlooked on initial examination. It takes particular experience to evaluate gastrointestinal biopsies from HIV-infected patients. PMID- 11473455 TI - The small blue cell dilemma associated with tamoxifen therapy. AB - CONTEXT: Several endometrial diseases, such as endometrial hyperplasia, endometrial carcinoma, and endometrial polyps, have been reported to be associated with tamoxifen administration. We recently observed a high incidence of distinctive small blue cells in Papanicolaou tests of women who had received tamoxifen treatment for breast carcinoma. OBJECTIVES: To define the characteristics of these small blue cells, to identify the patient population in which they are found, and to determine the clinical significance and possible etiology of these findings. DESIGN: A total of 154 Papanicolaou tests from 60 patients with a clinical history of tamoxifen therapy were reviewed retrospectively. RESULTS: Small blue cells were found in 40% of Papanicolaou tests from patients who received tamoxifen therapy. Patients with small blue cells in their Papanicolaou tests were an average of 9 years older at the time tamoxifen therapy was initiated than those without. Among the available follow-up surgical biopsies, no malignant diagnoses were made. CONCLUSIONS: We conclude that these distinctive small blue cells are found more frequently in older patients and most probably represent proliferative reserve cells of cervical/vaginal epithelium resulting from the estrogenic agonist effect of tamoxifen. More importantly, they are nonneoplastic in nature. PMID- 11473456 TI - Major pancreatic resections for chronic pancreatitis. AB - OBJECTIVE: Indications for major pancreatic resections have been expanded to include complicated chronic pancreatitis (CP). We assessed clinical findings and outcomes and evaluated histology in patients who had major pancreatic resections for CP. We also determined if histologic findings were associated with persistent postoperative pain. DESIGN: We reviewed charts and slides from 44 patients who underwent major pancreatic resections for CP between 1989 and 1999. RESULTS: The etiology for disease included alcohol (n = 15), hereditary (n = 5), idiopathic (n = 6), pancreas divisum (n = 3), stricture (n = 2), trauma (n = 2), systemic lupus erythematosus (n = 1), and unknown (n = 10). Patients included 20 men and 24 women; ages ranged from 22 to 76 years. Perioperative mortality and morbidity were 0% and 4.5%, respectively. Persistent pain was present in 25 (57%) of the 44 patients, and pain was encountered more frequently in patients with alcoholic pancreatitis (67%) versus other etiologies (52%), and in those who underwent Whipple/Beger or total resections (68%) versus distal or subtotal pancreatectomy (42%). Metaplastic changes were present in 14 cases, and ductal atypia was seen in 9 cases. No malignancies were found. Acinar necrosis and acute inflammation were seen more often in patients with persistent pain than in those who were pain free (P =.081). CONCLUSION: Major pancreatic resection for CP can be performed with low morbidity and mortality. This procedure relieves pain in nearly half the patients. There is a wide spectrum of histopathologic changes seen in CP, including ductal atypia and metaplastic changes. Acute exacerbations of CP identified histologically at the time of surgery and alcohol as etiology for CP may be associated more frequently with intractable pain. PMID- 11473457 TI - Adequate histologic sampling of breast core needle biopsies. AB - OBJECTIVE: To determine the degree of histologic sampling necessary for adequate examination of breast core needle biopsy specimens. DESIGN: The results of all breast core needle biopsies (11 and 14 gauge) with a diagnosis of atypical small acinar proliferation or atypical ductal hyperplasia and subsequent excisional biopsies, for a 50-month period were reviewed. Blocks of all cores were sectioned entirely in 8 slides to determine the amount of sectioning needed to detect these foci, and the results were correlated with those from the excision specimen. SETTING: Large community hospital practice. RESULTS: Of 3026 cases, 216 (7.1%) were diagnosed as atypical ductal hyperplasia or atypia not otherwise specified. Subsequent resections were available in 105 (49%) cases, and after review, 95 (92%) qualified as atypical ductal hyperplasia and 2 were determined to be atypical small acinar proliferations. The 2 small acinar proliferations were first detected on the second and fourth slides. Of the atypical ductal hyperplasia cases, 43% were detected on the first slide, 17% on the second, 23% on the third, 8% on the fourth, and 8% on the fifth. No lesions were initially detected after this level. Ductal carcinoma in situ was detected in the excision specimens from 1 case each of those detected initially on the fourth and fifth slides. CONCLUSION: Five sections of breast core needle biopsy specimens are necessary to ensure that all atypical small acinar proliferations and atypical ductal hyperplasia lesions are sampled. PMID- 11473458 TI - Pagetoid dyskeratosis is a frequent incidental finding in hemorrhoidal disease. AB - BACKGROUND: Pagetoid dyskeratosis is considered a selective keratinocytic response in which a small part of the normal population of keratinocytes is induced to proliferate. Pagetoid dyskeratosis has been found incidentally in the squamous epithelium of the skin in various locations and in the ectocervix in uterine prolapse. In cases in which these pale cells are conspicuous, there is a hazard of overdiagnosis. It has been suggested that friction is the most probable inductor of the lesion. To the best of our knowledge, pagetoid cells have not been reported in surgically resected hemorrhoids. OBJECTIVE AND DESIGN: We here describe the location of pagetoid dyskeratosis in the squamous epithelium of hemorrhoids and the incidence of this lesion in a group of 100 unselected patients surgically treated for hemorrhoidal disease. In addition to the conventional histologic method, special staining procedures and an immunohistochemical study of cytokeratins were performed in selected cases. RESULTS: Pagetoid dyskeratosis was found in 68 cases (68%) and was a prominent finding in 22 cases (22%). The cells of pagetoid dyskeratosis were strongly positive for high-molecular weight cytokeratin. These cells showed an immunohistochemical profile that was different from that of the surrounding squamous cells and indicative of premature keratinization. CONCLUSIONS: In hemorrhoidal disease, the cushions are susceptible to trauma as a result of prolapse. In this setting, friction may be the stimulus for the appearance of pagetoid dyskeratotic cells. These cells must be distinguished from the artifactual clear cells of the squamous epithelium, glycogen-rich cells, and koilocytes. The lesion must also be distinguished from extramammary Paget disease, pagetoid spread of carcinoma cells, pagetoid Bowen disease, and pagetoid melanoma. Pathologists should be familiar with the histologic features of pagetoid dyskeratosis in hemorrhoidectomy specimens to avoid misdiagnosis. Routine histologic study is usually adequate for recognizing this lesion. PMID- 11473459 TI - Evaluation of bone marrow specimens with acute myelogenous leukemia for CD34, CD15, CD117, and myeloperoxidase. AB - CONTEXT: Immunophenotyping of bone marrow (BM) specimens with acute myelogenous leukemia (AML) may be performed by flow cytometric (FC) or immunohistochemical (IH) techniques. Some markers (CD34, CD15, and CD117) are available for both techniques. Myeloperoxidase (MPO) analysis may be performed by enzyme cytochemical (EC) or IH techniques. OBJECTIVE: To determine the reliability of these markers and MPO by these techniques, we designed a study to compare the results of analyses of these markers and MPO by FC (CD34, CD15, and CD117), EC (MPO), and IH (CD34, CD15, CD117, and MPO) techniques. MATERIALS AND METHODS: Twenty-nine AMLs formed the basis of the study. These AMLs all had been immunophenotyped previously by FC analysis; 27 also had had EC analysis performed. Of the AMLs, 29 had BM core biopsies and 26 had BM clots that could be evaluated. The paraffin blocks of the 29 BM core biopsies and 26 BM clots were stained for CD34, CD117, MPO, and CD15. These results were compared with results by FC analysis (CD34, CD15, and CD117) and EC analysis (MPO). RESULTS: Immunodetection of CD34 expression in AML had a similar sensitivity by FC and IH techniques. Immunodetection of CD15 and CD117 had a higher sensitivity by FC analysis than by IH analysis. Detection of MPO by IH analysis was more sensitive than by EC analysis. There was no correlation of French-American-British (FAB) subtype of AML with CD34 or CD117 expression. Expression of CD15 was associated with AMLs with a monocytic component. Myeloperoxidase reactivity by IH analysis was observed in AMLs originally FAB subtyped as M0. CONCLUSIONS: CD34 can be equally detected by FC and IH techniques. CD15 and CD117 are better detected by FC analysis and MPO is better detected by IH analysis. PMID- 11473460 TI - Pseudosarcomatous myofibroblastic tumor and myosarcoma of the urogenital tract. AB - OBJECTIVE: Pseudosarcomatous myofibroblastic tumors (PMTs) of the urogenital tract are rare but distinctive lesions. Despite their benign behavior, they are frequently misinterpreted as leiomyosarcomas and rhabdomyosarcomas in preoperative biopsies and even in resected specimens because of their atypical spindle-cell features. Precise diagnosis of PMTs is important to avoid unnecessary radical therapy. We analyzed urogenital myoid tumors to clarify which of their characteristics are useful for the differential diagnosis. METHODS: We evaluated 7 urogenital myoid tumors consisting of 3 PMTs, 2 leiomyosarcomas, and 2 rhabdomyosarcomas. We studied the expression of various immunohistochemical muscle-cell markers including desmin, muscle-specific actin, alpha-smooth muscle actin, high-molecular-weight caldesmon, and myogenin. RESULTS: Desmin, muscle specific actin, and alpha-smooth muscle actin were noted variably in all tumor types, whereas high-molecular-weight caldesmon was expressed only in leiomyosarcomas, and myogenin was expressed only in rhabdomyosarcomas. CONCLUSION: High-molecular-weight caldesmon and myogenin are useful for differentiating urogenital PMTs from myosarcomas. PMID- 11473461 TI - Cytokeratin subset immunostaining in rectal adenocarcinoma and normal anal glands. AB - CONTEXT: A large percentage of cases of perianal Paget disease are associated with an internal cancer, most commonly rectal adenocarcinoma. Immunostains for cytokeratin 7, cytokeratin 20, and gross cystic disease fluid protein 15 are useful in identifying cases associated with rectal adenocarcinoma. The Paget cells and rectal adenocarcinoma cells of these lesions typically have a cytokeratin 7(+)/cytokeratin 20(+)/gross cystic disease fluid protein 15(-) immunophenotype. It is not known whether rectal adenocarcinoma unassociated with perianal Paget disease has the same cytokeratin profile as rectal adenocarcinoma associated with perianal Paget disease. OBJECTIVE: To evaluate the immunohistochemical cytokeratin 7 and 20 profile of resected rectal adenocarcinoma unassociated with perianal Paget disease as well as that of normal anal glands from hemorrhoidectomy specimens. DESIGN: We performed immunohistochemistry for cytokeratins 7 and 20 on tissues from 30 cases of rectal adenocarcinoma unassociated with perianal Paget disease and 12 hemorrhoidectomy specimens from 12 cases with normal anal glands. We defined positive staining as any immunoreactivity within the neoplastic cells. RESULTS: Twenty-six of 30 cases of rectal adenocarcinoma (87%) had a cytokeratin 7(-)/cytokeratin 20(+) immunophenotype, similar to the immunophenotype of cases of nonrectal large intestine adenocarcinoma. In 4 cases (13%), neoplastic cells coexpressed cytokeratins 7 and 20. Anal glands stained strongly for cytokeratin 7 but were negative for cytokeratin 20 in all cases, and the anal transitional zone mucosa had a similar immunophenotype. CONCLUSIONS: Rectal adenocarcinoma unassociated with perianal Paget disease has a cytokeratin profile similar to that of nonrectal large intestine adenocarcinoma. These data suggest that rectal adenocarcinoma unassociated with perianal Paget disease has a different cytokeratin profile than rectal adenocarcinoma associated with perianal Paget disease. PMID- 11473462 TI - Primary plasmacytoma of the breast. AB - We describe a solitary extramedullary plasmacytoma of the breast in a 37-year-old woman. No other involvement was detected in the bone marrow or in any other site during a 15-month follow-up period. Extramedullary plasmacytomas of the breast are extremely rare, especially those that are not associated with multiple myeloma. We review the histologic features of the previously reported cases with an emphasis on differential diagnosis and the difficulties encountered in arriving at the correct diagnosis in frozen sections. PMID- 11473463 TI - Adult extrarenal Wilms tumor occurring in the uterus. AB - Five previous cases of extrarenal Wilms tumor (EWT) occurring in the uterus have been reported. The oldest patient was 22 years. We report a case of uterine EWT occurring in a 42-year-old woman. Histologically, there was typical triphasic differentiation, including epithelial, blastemal, and mesenchymal elements. The important differential diagnosis in this age group, the malignant mixed mullerian tumor, is excluded by the absence of glomeruloid structures and primitive tubules. The exact histogenesis of EWT is unknown but most likely relates to the presence of nephrogenic rests occurring in the female genital tract. PMID- 11473464 TI - Metastatic carcinomatous cirrhosis and hepatic hemosiderosis in a patient heterozygous for the H63D genotype. AB - A 38-year-old woman had a mastectomy for infiltrating ductal carcinoma of the breast 3 years before her last admission and had received chemotherapy for known liver metastases. She developed the rapid onset of liver failure with portal hypertension and died in a hospice. Autopsy revealed macronodular cirrhosis of the liver secondary to metastatic carcinoma of the breast with associated florid fibrosis. This rare lesion, previously called metastatic carcinomatous cirrhosis, was also found, in this case, to have marked hepatic hemosiderosis, and analysis of the patient's DNA showed heterozygosity for the H63D genotype. The possibility of cirrhosis-associated hemosiderosis secondary to an iron metabolism abnormality associated with the H63D mutation of the HFE gene is proposed. Computed tomographic scans showed the development of cirrhosis during the 3-month period before the patient's last admission and suggested the possibility of a postnecrotic type origin. PMID- 11473465 TI - Endometrial stromal sarcoma of the rectosigmoid colon arising in extragonadal endometriosis and revealed by portal vein thrombosis. AB - Malignant transformation is an infrequent complication of endometriosis. The ovary is the primary site in 76% of cases, and extragonadal sites are identified in 24%. Endometrioid carcinoma is the most common histologic type; sarcoma is very rare. We report a case of low-grade endometrial stromal sarcoma of the rectosigmoid colon presenting with epigastric pain due to portal vein thrombosis. This tumor arose from extragonadal endometriosis in a 61-year-old woman and was treated by surgical resection. The main differential diagnosis of this unusual colonic neoplasm includes primary mesenchymal tumors, such as gastrointestinal stromal tumors. PMID- 11473466 TI - Fine-needle aspiration biopsy features in a case of giant cell fibroblastoma of the chest wall. AB - Giant cell fibroblastoma is an unusual tumor of childhood, primarily occurring in the superficial soft tissues. We describe the fine-needle aspiration biopsy features of a case of giant cell fibroblastoma of the chest wall in a 3-year-old child. The aspirates comprised bland spindle to oval cells entrapped in a metachromatic matrix, accompanied by rare multinucleated giant cells with wreathlike nuclei. Although we were unable to render a definitive diagnosis on fine-needle aspiration biopsy, surgical resection of the mass established the diagnosis of giant cell fibroblastoma. We review the distinctive cytologic features of some common soft tissue tumors arising in this age group that may give rise to a diagnostic conundrum on fine-needle aspiration biopsy. PMID- 11473467 TI - Inflammatory pseudotumor of the submandibular gland: report of a case presenting with autoimmune disease-like clinical manifestations. AB - We report a rare case of inflammatory pseudotumor arising in the submandibular gland, which presented with autoimmune disease-like clinical manifestations. A 70 year-old Japanese man developed masses in both submandibular regions. Laboratory tests revealed polyclonal hypergammaglobulinemia, high titers of antinuclear antibody, and a positive thyroid test. Histologically, the lesion was composed of multiple nodules separated by thick fibrous bands and contained a few atrophic lymphoid follicles and residual ductal structures. At higher magnification, the nodules contained numerous mature plasma cells mixed with myofibroblasts, lymphocytes, and histiocytes. Occasionally, the myofibroblasts were arranged in poorly formed fascicles and in a storiform pattern. Polymerase chain reaction analysis failed to demonstrated the rearrangement of the immunoglobulin heavy chain gene. The patient was free of disease after 72 months follow-up. Marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) type arising from salivary glands occasionally showed prominent plasma cell differentiation. The present case indicates that inflammatory pseudotumor should be added to the list of different diagnoses for mucosa-associated lymphoid tissue-type lymphoma of the salivary glands. PMID- 11473468 TI - Nasopharyngeal gangliocytic paraganglioma. AB - Gangliocytic paraganglioma (GP) is a rare neoplasm described almost exclusively in the gastrointestinal tract, especially the periampullary region. However, several examples have been reported at various sites, including the stomach, jejunum, and appendix. Herein we report a case of GP involving the nasopharynx. To our knowledge, this is the first report of GP at this site. A 44-year-old woman presented with headaches and symptoms of fullness and pressure related to mass effect. An initial endoscopic biopsy was followed by surgical excision of the nasopharyngeal mass. The triphasic tumor fulfilled the morphologic and immunohistochemical criteria for GP. The histogenesis of GP is uncertain, and the current belief is that it arises from the embryonic ventral pancreas. This concept is based largely on the location of most cases, which is along the embryologic migration route of the ventral pancreas, as well as the expression of pancreatic polypeptide by the tumor. The nasopharyngeal location of our case clearly refutes the pancreatic origin of GP. We propose that the tumor probably arises from totipotential adult stem cells, which in the right microenvironment differentiate along nonnative cell lineages. PMID- 11473469 TI - Metastatic prostatic adenocarcinoma within a primary solid papillary carcinoma of the male breast. AB - We report the case of a 78-year-old man who developed a breast mass 12 months after hormonal therapy for palliation of prostatic adenocarcinoma. On histologic and immunohistochemical examination, the breast tumor revealed a unique collision tumor composed of metastatic prostatic adenocarcinoma and solid papillary breast carcinoma. PMID- 11473470 TI - Cytokeratin-positive, CD45-negative primary centroblastic lymphoma of the adrenal gland: a potential for a diagnostic pitfall. AB - We report a case of cytokeratin-positive, CD45-negative primary polymorphic centroblastic lymphoma of the adrenal gland. Additional immunostaining, which demonstrated positivity for CD20 and kappa light chain, as well as detection of the monoclonal rearrangement of the immunoglobulin heavy chain gene, helped to establish the diagnosis of lymphoma and to rule out an initially favored diagnosis of poorly differentiated carcinoma. PMID- 11473471 TI - Vibrio vulnificus septicemia in a patient with the hemochromatosis HFE C282Y mutation. AB - Vibrio vulnificus is an extremely invasive gram-negative bacillus found in marine waters that causes overwhelming bacteremia and shock that is associated with high mortality. Impaired iron metabolism has been implicated in the susceptibility to V vulnificus bacterial infections. We report a case of fatal V vulnificus sepsis in a 56-year-old man who died within 1 to 3 days after consuming raw seafood. At autopsy, he was found to have micronodular cirrhosis and iron overload. Postmortem genetic analysis revealed the presence of the hemochromatosis gene (HFE) C282Y mutation. To our knowledge, this is this first documented fatal case of V vulnificus infection in a patient proven to carry the HFE C282Y mutation. Because this patient was heterozygous for the major hereditary hemochromatosis mutation and was not previously diagnosed with clinical iron overload, the spectrum of clinical susceptibilities to V vulnificus infection may include carriers of the C282Y mutation. PMID- 11473472 TI - Colonic lymphoid hyperplasia in melanosis coli. AB - We describe the case of a patient with Rett syndrome, a syndrome characterized by progressive infant encephalopathy, developmental delay, dementia, autism, ataxia, microcephaly, spastic paraparesis, and autonomic neuropathy with constipation. At colonoscopy, multiple foci of tiny white, sessile, polypoid lesions were seen throughout the colon and rectum, mimicking the appearances of small hyperplastic or adenomatous polyps, associated with generalized melanosis coli. This is the first case to our knowledge describing melanosis coli in a patient with Rett syndrome. As melanosis pigment deposition is characteristically not seen in lymphoid tissue, the lymphoid tissue was identifiable at endoscopy as multiple white nodules mimicking generalized colonic polyposis throughout the colon. We discuss the likely mechanisms of lymphoid hyperplasia and coexistent melanosis coli in Rett syndrome. PMID- 11473473 TI - Pathologic quiz case: an unusual cerebellopontine angle tumor. PMID- 11473474 TI - Pathologic quiz case: a 48 year-old woman with a dural-based intracranial tumor. PMID- 11473475 TI - Pathologic quiz case: rapidly growing expansile lesion of the left jaw in a 15 year-old boy. PMID- 11473476 TI - Pathologic quiz case: a 77-year-old woman with obstructive hydrocephalus. PMID- 11473477 TI - Multilocular peritoneal inclusion cyst (benign cystic mesothelioma). PMID- 11473478 TI - Platelet storage pool deficiency of alpha and delta granules. PMID- 11473479 TI - Acinar cell carcinoma of the pancreas. PMID- 11473484 TI - Binding and uptake of chylomicron remnants by primary and THP-1 human monocyte derived macrophages: determination of binding proteins. AB - The binding and uptake of chylomicron remnants by human macrophages was studied in order to resolve paradoxical observations that have described the putative mechanisms by which postprandial lipoproteins induce foam cell formation. Chylomicron remnants bound to human monocyte-derived macrophages (HMMs) and to the transformed monocytic cell line THP-1 with high affinity (Kd of approx. 5.5 microg of chylomicron remnant protein/ml). Binding was found to be saturable for both cell types, and was strongly inhibited in the presence of unlabelled chylomicron remnants. Ligand blot studies with colloidal-gold-labelled chylomicron remnants identified two cell surface binding sites on both HMMs and THP-1 cells, with molecular masses of approx. 128 kDa and 43 kDa. The high molecular-mass binding site was found to be the low-density lipoprotein (LDL) receptor, based on the strong inhibition of chylomicron remnant binding in the presence of unlabelled LDL, Fab2 antibody fragments to the LDL receptor or calcium chelators. Competition studies suggested that, in HMMs, the LDL receptor appeared to facilitate approximately half of the total chylomicron remnant uptake. In contrast, the LDL receptor was not significantly involved in macrophage uptake of chylomicron remnants by THP-1 cells. The identity of the 43 kDa binding site is presently unknown, but, importantly, expression was not inhibited as a consequence of sterol loading, which was induced by incubating HMMs and THP-1 cells with 25-hydroxycholesterol. In contrast, the expression of the LDL receptor was substantially attenuated following lipid loading. Collectively, our data suggest that, while the macrophage LDL receptor can bind chylomicron remnants and facilitate uptake in non-lipid-loaded HMMs, other sterol insensitive sites are responsible for the unabated uptake of chylomicron remnants by macrophages. We propose that the 43 kDa macrophage chylomicron remnant binding protein may be a candidate for the sterol loading of macrophages. PMID- 11473485 TI - Biophysical effects of pore mutations of ROMK1. AB - Potassium channels are ubiquitous, being present in all living organisms. These proteins share common structural elements, which confer common functional features. In general, all K+ channels have a high selectivity for K+, and are blocked by cations of similar dimensions, such as Cs+ and Ba2+. Mutations in the pore region tend to lead to either the total loss of function or K+ selectivity. We have made mutations to one of the most highly conserved residues of the pore, glycine-143, of the inward rectifier ROMK1 (Kir1.1), and examined the resulting channel properties in the Xenopus oocyte expression system with a two-electrode voltage clamp. Mutations G143A and G143R resulted in failure to express functional channels. Co-injection of wild-type ROMK1 cRNA with these mutants led to rescue of channel function, which was different from wild-type ROMK1. In both mutants, the sensitivity to Ba2+ and Cs+ was increased, the rate of onset of block by Ba2+ was enhanced, and the selectivity to potassium was reduced. Whereas the crystallographic evidence shows that cations bind to the carbonyl backbone of the pore-lining residues, the present results indicate that the side chains of these amino acids, which face away from the pore lining, also affect permeation. PMID- 11473486 TI - Differences in lymph drainage between swollen and non-swollen regions in arms with breast-cancer-related lymphoedema. AB - Recent research indicates that the pathophysiology of breast-cancer-related lymphoedema (BCRL) is more complex than simple axillary lymphatic obstruction as a result of the cancer treatment. Uneven distribution of swelling (involvement of the mid-arm region is common, but the hand is often spared) is puzzling. Our aim was to test the hypothesis that local differences in lymphatic drainage contribute to the regionality of the oedema. Using lymphoscintigraphy, we measured the removal rate constant, k (representing local lymph flow per unit distribution volume, VD), for 99mTc-labelled human immunoglobulin G in the oedematous proximal forearm, and in the hand (finger web) in women in whom the hand was unaffected. Tracer was injected subcutaneously, and the depot plus the rest of the arm was monitored with a gamma-radiation camera for up to 6 h. VD was assessed from image width. Contralateral arms served as controls. k was 25% lower in oedematous forearm tissue than in the control arm (BCRL, -0.070+/-0.026% x min(-1); control, -0.093+/-0.028% x min(-1); mean+/-S.D.; P=0.012) and VD was greater. In the non-oedematous hand of the BCRL arm, k was 18% higher than in the control hand (BCRL, -0.110+/-0.027% x min(-1); control, -0.095+/-0.028% x min( 1); P=0.057) and 59% higher than forearm k on the BCRL side (P=0.0014). VD did not differ between the hands. Images of the BCRL arm following hand injection showed diffuse activity in the superficial tissues, sometimes extending almost to the shoulder. A possible interpretation is that the hand is spared in some patients because local lymph flow is increased and diverted along collateral dermal routes. The results support the hypothesis that regional differences in surviving lymphatic function contribute to the distribution of swelling. PMID- 11473487 TI - Sympathetic and reflex abnormalities in heart failure secondary to ischaemic or idiopathic dilated cardiomyopathy. AB - Congestive heart failure (CHF) is characterized by a sympathetic activation and a baroreflex impairment whose degree is directly related to the clinical severity of the disease. However, whether these abnormalities vary according to the ischaemic or idiopathic dilated nature of the CHF state has not been conclusively documented. In patients with a clinically stable, chronic CHF state in New York Heart Association functional class II and III, due either to ischaemic heart disease (IHD; n=22, age 60.3+/-2.4 years, means+/-S.E.M.) or to idiopathic dilated cardiomyopathy (IDC; n=20, age 58.9+/-2.8 years), and in 30 age-matched controls, we measured arterial blood pressure (using a Finapres device), heart rate (by electrocardiogram) and postganglionic muscle sympathetic nerve traffic (by microneurography) at rest and during baroreceptor manipulation induced by the vasoactive drug-infusion technique. Blood pressure values were not significantly different in CHF patients and controls. Compared with controls, heart rate was similarly increased and left ventricular ejection fraction (by echocardiography) similarly reduced in CHF patients with IHD or IDC. Muscle sympathetic nerve traffic was significantly greater in CHF patients than in controls, and did not differ between patients with IHD or IDC (67.3+/-4.2 and 67.8+/-3.8 bursts/100 heart beats respectively). This was also the case for the degree of baroreflex impairment. These data show that CHF states due to IHD or to IDC are characterized by a similar degree of peripheral sympathetic activation and by a similar impairment of the baroreflex function. Thus the neuroadrenergic and reflex abnormalities characterizing CHF are independent of its aetiology. PMID- 11473488 TI - Effects of hormone replacement therapy and high-impact physical exercise on skeletal muscle in post-menopausal women: a randomized placebo-controlled study. AB - An age-related decline in muscle performance is a known risk factor for falling, fracture and disability. In women, a clear deterioration is observed from early menopause. The effect of hormone replacement therapy (HRT) in preserving muscle performance is, however, unclear. This trial examined the effects of a 12-month HRT and high-impact physical exercise regimen on skeletal muscle in women in early menopause. A total of 80 women aged 50-57 years were assigned randomly to one of four groups: exercise (Ex), HRT, exercise+HRT (ExHRT) and control (Co). The exercise groups participated in a high-impact training programme. The administration of HRT (oestradiol/noretisterone acetate) or placebo was carried out double-blind. Knee extension torque and vertical jumping height were evaluated. Lean tissue cross-sectional area (LCSA) and the relative proportion of fat within the muscle compartment were measured for the quadriceps and lower leg muscles. The ExHRT group showed significant increases in knee extension torque (8.3%) and vertical jumping height (17.2%) when compared with the Co group ( 7.2%). Vertical jumping height also increased after HRT alone (6.8%). The LCSA of the quadriceps was increased significantly in the HRT (6.3%) and ExHRT (7.1%) groups when compared with the Ex (2.2%) and Co (0.7%) groups. Lower leg LCSA was also increased in the ExHRT group (9.1%) when compared with the Ex (3.0%) and Co (4.1%) groups. In addition, the increase in the relative proportion of fat in the quadriceps in the Co group (16.6%) was significant compared with those in the HRT (4.9%) and ExHRT (-0.6%) groups. Thus, in post-menopausal women, muscle performance, muscle mass and muscle composition are improved by HRT. The beneficial effects of HRT combined with high-impact physical training may exceed those of HRT alone. PMID- 11473489 TI - A comparative study of the adsorption of amino acids on to calcium minerals found in renal calculi. AB - To assess the binding of individual amino acids to the principal calcium minerals found in human kidney stones, the adsorption of 20 amino acids on to calcium oxalate monohydrate, CaHPO4*2H2O, Ca3(PO4)2 and Ca5(PO4)3OH crystals was determined over the physiological urinary pH range (pH 5-8) in aqueous solutions. All amino acids adsorbed most strongly at pH 5, and this decreased in all cases as the pH was increased. The amino acids which adsorbed most strongly were aspartic acid, glutamic acid and gamma-carboxyglutamic acid, with the last displaying the strongest affinity. All amino acids bound more avidly to calcium oxalate monohydrate than to any of the phosphate minerals. Adsorption on to CaHPO4*2H2O was generally higher than for Ca3(PO4)2 and Ca5(PO4)3OH, for which all amino acids, with the exception of gamma-carboxyglutamic acid, had only a weak affinity. The binding affinity of these acids is thought to be due to their zwitterions being able to adopt conformations in which two carboxyl groups, and possibly the amino group, can interact with the mineral surface without further rotation. The strong binding affinity of di-and tri-carboxylic acids for calcium stone minerals indicates that proteins rich in these amino acids are more likely to play a functional role in stone pathogenesis than those possessing only a few such residues. These findings, as well as the preferential adsorption of the amino acids for calcium oxalate monohydrate rather than calcium phosphate minerals, have ramifications for research aimed at discovering the true role of proteins in stone formation and for potential application in the design of synthetic peptides for use in stone therapy. PMID- 11473490 TI - Novel insight into the pathophysiology of breast-cancer-related lymphoedema. PMID- 11473491 TI - Counteracting sarcopenia in post-menopausal women: do hormones and strength training accomplish the task? PMID- 11473492 TI - Dilution and redistribution effects of rapid 2-litre infusions of 0.9% (w/v) saline and 5% (w/v) dextrose on haematological parameters and serum biochemistry in normal subjects: a double-blind crossover study. AB - Although hypoalbuminaemia after injury may result from increased vascular permeability, dilution secondary to crystalloid infusions may contribute significantly. In this double-blind crossover study, the effects of bolus infusions of crystalloids on serum albumin, haematocrit, serum and urinary biochemistry and bioelectrical impedance analysis were measured in healthy subjects. Ten male volunteers received 2-litre infusions of 0.9% (w/v) saline or 5% (w/v) dextrose over 1 h; infusions were carried out on separate occasions, in random order. Weight, haemoglobin, serum albumin, serum and urinary biochemistry and bioelectrical impedance were measured pre-infusion and hourly for 6 h. The serum albumin concentration fell in all subjects (20% after saline; 16% after dextrose) by more than could be explained by dilution alone. This fall lasted more than 6 h after saline infusion, but values had returned to baseline 1 h after the end of the dextrose infusion. Changes in haematocrit and haemoglobin were less pronounced (7.5% after saline; 6.5% after dextrose). Whereas all the water from dextrose was excreted by 2 h after completion of the infusion, only one-third of the sodium and water from the saline had been excreted by 6 h, explaining its persistent diluting effect. Impedances rose after dextrose and fell after saline (P<0.001). Subjects voided more urine (means 1663 and 563 ml respectively) of lower osmolality (means 129 and 630 mOsm/kg respectively) and sodium content (means 26 and 95 mmol respectively) after dextrose than after saline (P<0.001). While an excess water load is excreted rapidly, an excess sodium load is excreted very slowly, even in normal subjects, and causes persistent dilution of haematocrit and serum albumin. The greater than expected change in serum albumin concentration when compared with that of haemoglobin suggests that, while dilution is responsible for the latter, redistribution also has a role in the former. Changes in bioelectrical impedance may reflect the electrolyte content rather than the volume of the infusate, and may be unreliable for clinical purposes. PMID- 11473493 TI - To bolus or not to bolus--is that the question? PMID- 11473494 TI - Acute mental stress elicits delayed increases in circulating inflammatory cytokine levels. AB - The influence of acute mental stress on cardiovascular responses and concentrations of inflammatory cytokines up to 2 h later was assessed in 12 subjects exposed to stress and in eight control subjects. Beat-by-beat recordings of finger blood pressure and heart rate were made at rest and during two behavioural tasks (colour-word interference and mirror tracing). Blood was drawn after adaptation and at 45 min and 2 h after the tasks, and assayed for interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-alpha), interleukin-1 receptor antagonist (IL-1Ra), C-reactive protein (CRP) and haematocrit. Saliva was sampled periodically and assayed for free cortisol. The tasks were rated as stressful by the participants. The stress group showed significant increases in systolic and diastolic blood pressure (mean rises of 16.4+/-12.3 and 12.6+/-6.9 mmHg respectively) and heart rate (5.39+/-5.3 beats/min); these values returned to baseline during the recovery period. The IL-6 concentration was increased by 56% at 2 h after the tasks (P<0.05), while IL-1Ra was increased by 12.3% (P<0.01). No changes in cardiovascular variables or cytokine concentrations were observed in the control subjects, and haematocrit did not change. The magnitude of blood pressure responses during tasks was correlated positively with the IL-6 concentration after 45 min (r=0.70, P<0.05), and with the IL-1Ra concentration after 2 h (r=0.63, P<0.05). Increases in TNF-alpha after 2 h were correlated with heart rate responses to tasks (r=0.66, P<0.05). Associations between IL-6 and IL 1Ra concentrations were also recorded. This study indicates that inflammatory cytokines respond to acute mental stress in humans with delayed increases, and suggest that individual differences in cytokine responses are associated with sympathetic reactivity. PMID- 11473495 TI - Psychological stress and the inflammatory response system. PMID- 11473496 TI - In vivo inhibition of renal 11beta-hydroxysteroid dehydrogenase in the rat stimulates collecting duct sodium reabsorption. AB - In order to test the proposal that the aldosterone specificity of mineralocorticoid receptors in the collecting duct depends on inactivation of glucocorticoids by the enzyme 11beta-hydroxysteroid dehydrogenase (11beta-HSD), we have assessed the effect of pharmacological inhibition of 11beta-HSD on collecting duct Na+ reabsorption in vivo. Adrenalectomized rats (n=14) were infused intravenously with high-dose corticosterone, and late-distal tubules were perfused orthogradely with artificial tubular fluid containing [14C]inulin and 22Na; urinary recoveries of the radioisotopes were monitored. Half of the rats received intravenous carbenoxolone to inhibit renal 11beta-HSD activity. The urinary recovery of [14C]inulin was complete in both groups of animals (101+/-2% versus 101+/-3%), but the recovery of 22Na was lower in carbenoxolone-treated rats (34+/-5%) than in the corticosterone-alone group (54+/-4%, P<0.01). These data, which provide the first demonstration of enhanced Na+ reabsorption in the distal nephron during inhibition of renal 11beta-HSD in vivo, strongly support the proposal that 11beta-HSD normally prevents endogenous glucocorticoid from exerting mineralocorticoid-like effects. PMID- 11473497 TI - Diagnosing coeliac disease by rectal gluten challenge: a prospective study based on immunopathology, computerized image analysis and logistic regression analysis. AB - The purpose of this study was to evaluate the use of rectal gluten challenge in the diagnosis of coeliac disease. A total of 103 patients with features suggestive of this diagnosis were prospectively enrolled into the study; a diagnosis of coeliac disease was based on strictly defined criteria used in judging the proximal jejunal biopsy. On that basis, 45 out of the 103 patients were deemed to have coeliac disease. A slurry of gluten powder in physiological saline was introduced into the rectum, and biopsies taken before and at 2 h or 4 h after the challenge were examined immunohistochemically by computerized image analysis. Cell counts were analysed by logistic regression, and the best equations were obtained for each challenge group. The 2 h challenge yielded diagnostic sensitivity and specificity of 69.6% and 78.6% respectively. The 4 h challenge provided sensitivity and specificity of 100% and 100% respectively. These results were compared with other clinical diagnostic predictors,including anti-endomysial antibodies, which yielded diagnostic sensitivity and specificity of 70% and 98% respectively. It is concluded that a 4 h rectal challenge is a highly sensitive means of identifying gluten-sensitized individuals, and would be of particular value in cases showing negative antibody screening or equivocal biopsy appearances. PMID- 11473498 TI - Why do smokers have higher plasma fibrinogen levels than non-smokers? PMID- 11473499 TI - Major depressive disorder: longitudinal symptomatic structure, relapse and recovery. PMID- 11473500 TI - Selective serotonin reuptake inhibitors and violence: a review of the available evidence. AB - OBJECTIVE: To provide a clinically useful analysis of the relationship between selective serotonin reuptake inhibitors (SSRIs), in particular fluoxetine and violent or suicidal behaviour. METHOD: All published papers on Medline and other databases linking serotonin, SSRIs and aggression were reviewed. RESULTS: A small proportion of patients treated with SSRIs may become akathisic and others may show increases in anxiety in the initial phase of treatment, but no increased susceptibility to aggression or suicidality can be connected with fluoxetine or any other SSRI. In fact SSRI treatment may reduce aggression, probably due to positive effects on the serotonergic dysfunction that is implicated in aggressive behaviour directed towards oneself or others. CONCLUSION: In the absence of convincing evidence to link SSRIs causally to violence and suicide, the recent lay media reports are potentially dangerous, unnecessarily increasing the concerns of depressed patients who are prescribed antidepressants. PMID- 11473501 TI - Forty years of antidepressant drug trials. AB - OBJECTIVE: This study was conducted to investigate whether the quality of antidepressant drug trials has improved during the last 40 years. METHOD: A sample of 314 randomized clinical trials published between 1962 and 1998 was analysed. RESULTS: From 1962 to 1970 the median number of patients per trial was 56 (range 24-137), from 1971 to 1980 was 50 (10-211) and from 1981 to 1990 was 51 (20-314). In the last 8 years the median sample size increased to 100 patients (20-1002). Trials had a median duration of 4 weeks in the first two decades of publication, and a median duration of 6 weeks in the following two decades. Patient selection criteria have become increasingly sophisticated and the median number of efficacy measures increased in the last 4 decades from 1 to 4. CONCLUSION: Stringent selection criteria and sophisticated outcome assessment tend to exclude typical patients from randomized controlled trials and made it more difficult to follow many patients in the long term. PMID- 11473502 TI - Clinical significance and comorbidity of subthreshold depression and anxiety in the community. AB - OBJECTIVE: This report examines: 1) the magnitude of co-occurrence of threshold and subthreshold-level depression and anxiety in the community, and 2) the relationship between comorbidity and the diagnostic level of depression and anxiety and their clinical correlates. METHOD: A community sample of 591 subjects was interviewed prospectively five times across 15 years from the ages of 20-35. The diagnostic interview allowed the assignment of diagnoses according to DSM-III criteria and operational definitions of subthreshold syndromes. RESULTS: 1) Comorbidity between depression and anxiety was more frequent when one syndrome reached threshold level; 2) comorbidity at both the threshold and subthreshold diagnostic levels was associated with symptom severity, disability and treatment, whereas the diagnostic level was associated with disability and suicidal attempts. CONCLUSION: The systematic association between comorbidity across diagnostic threshold levels of anxiety and depression with clinical correlates suggests the importance of a more dimensional approach to their classification. PMID- 11473503 TI - Moclobemide vs. imipramine in bipolar depression: a multicentre double-blind clinical trial. AB - OBJECTIVE: To determine the relative efficacy, tolerability and risk of precipitating mania of moclobemide and imipramine in the treatment of bipolar depression. METHOD: A randomized, double-blind, parallel group, multicentre study of moclobemide (MCB) (450-750 mg daily) and imipramine (IMI) (150-250 mg daily) in 21 centres in nine countries; 156 patients (65 males, 91 females) aged 18-65 with bipolar depression (17-item Hamilton Depression Rating Scale (HAMD) score chi16) participated. Clinical status was assessed using standardized rating scales before treatment and at 1,2,3,4,6 and 8 weeks. The data were analysed on an intention to treat basis with the last observation carried forward. RESULTS: In the MCB group, the mean HAMD fell from 23.0 to 13.1, in the IMI group it fell from 22.5 to 9.5; the mean score on the Montgomery-Asberg Depression Rating Scale (MADRS) fell from 29.5 to 16.3 on MCB and from 29.2 to 11.6 on IMI. There were no statistically significant differences between the two groups on any efficacy measures. Anticholinergic side-effects were three times more common with IMI than MCB and weight gain was also greater on IMI. Two patients (3.7%) on MCB and six patients (11%) on IMI were withdrawn because of manic symptoms, with manic symptoms occurring earlier on IMI, although these differences did not reach statistical significance. CONCLUSION: No differences in efficacy were detected between MCB and IMI in the treatment of bipolar depression. The data suggests that MCB is less likely than IMI to precipitate mania. PMID- 11473504 TI - The epidemiology of perimenstrual psychological symptoms. AB - OBJECTIVE: The purpose of this paper is to examine the prevalence, clinical significance and comorbidity of perimenstrual symptoms/syndrome (PERI-MS) in a community sample of women. METHOD: Premenstrual and menstrual physical and psychological symptoms were assessed in a prospective longitudinal study of a representative community cohort of women (N=299) who were interviewed five times between the ages of 21 and 35 years. RESULTS: When classified according to the severity of distress, the prevalence rates were 8.1% for severe and 13.6% for moderate PERI-MS, respectively. Clinical indicators of severity were all greater among PERI-MS women with than without depressed mood. CONCLUSION: The results of this study suggest that irritability, nervousness and tension irrespective of the presence of concomitant depressed mood are core elements of the perimenstrual syndrome. PMID- 11473505 TI - Long-term risk factors for suicide mortality after attempted suicide--findings of a 14-year follow-up study. AB - OBJECTIVE: To determine the risk of suicide over a 14-year follow-up period, and to investigate the long-term risk factors for suicide using survival analysis. METHOD: Data were collected on all unselected deliberate self-poisoning patients (n=1018) treated during 1983 in the emergency unit of Helsinki University Central Hospital. RESULTS: By the end of the 14 -year follow-up period 222 (21.7%) of these patients had died. Sixty-eight (6.7%) had committed suicide; 44 (9.2%) men and 24 (4.5%) women. The long-term risk factors for suicide were male sex, previous psychiatric treatment, previous suicide attempts, somatic disease and a self-reported 'wish to die' motive for the index suicide attempt. CONCLUSION: The essential risk factors for suicide were being male and having previous suicide attempts. In addition, history of earlier psychiatric treatment, presence of somatic disease and genuine intent to die in the index suicide attempt suggest that the long-term risk has remained high for over a decade. The findings emphasize the need for long-term planning and treatment of suicide attempters met in the emergency room of general hospitals. PMID- 11473506 TI - Prevalence, incidence and prospective risk factors for eating disorders. AB - OBJECTIVE: To examine the prevalence, incidence and prospective risk factors for eating disorders (ED) among young adult females. METHOD: Using a prospective design, a randomly selected sample of 1157 females (18-30 years) from the general population was examined with questionnaires for establishing ED diagnoses, self esteem, body concern, coping and perceived social support. RESULTS: The prevalence of ED was 3.2% and the 2-year first-time incidence was 0.0105 (n=8). Subjects in the extended incidence group (n=34) reported significantly lower self esteem and perceived social support, and higher body concern and relative use of escape-avoidance coping, at the onset of the study in 1997, compared to controls (n=643). Furthermore, they reported a significant increase in body concern and relative use of escape-avoidance coping, and a significant decrease in self esteem compared to controls from 1997 to 1999. CONCLUSION: These factors may be considered as risk factors for later development of ED among young adult women. PMID- 11473507 TI - Bone mineral content and bone mineral density in adolescent girls with anorexia nervosa--a longitudinal study. AB - OBJECTIVE: Total body and lumbar spine bone mineral density (BMD-TB, BMD-L) and total body bone mineral content (BMC-TB) were measured to establish the course of bone demineralization in anorexia nervosa and the clinical factors influencing BMC-TB and BMD changes during treatment. METHOD: Forty-two girls with DSM III-R anorexia nervosa, age 14.7+/-2.4 years. BMC-TB, BMD-TB and BMD-L were measured in approximately 7-month intervals for 27.8+/-4.1 months using DXA. RESULTS: Despite nutritional improvement, there was an initial decrease of BMD-L, and no change in BMC-TB and BMD-TB. an increase in BMC-TB and BMD was observed after approx. 21 months from the beginning of the study. CONCLUSION: The improvement in BMC-TB and BMD was related to changes in nutritional status and was significantly marked in younger patients, with earlier anorexia onset and before menarche. PMID- 11473508 TI - Season of birth in schizophrenia and affective psychoses in Western Australia 1916-61. AB - OBJECTIVE: This study examines seasonality birth effects in schizophrenia and affective psychoses in the southern hemisphere, given possible confounding of age incidence effects with winter birth peaks in northern hemisphere data. METHOD: Distributions of births by season, quarter and month for individuals born in Western Australia 1916-61 with a diagnosis of schizophrenia (N=2284), affective psychoses (N=3236), and neurotic depression (N=4869) on the statewide mental health register were compared with distributions for the general population. RESULTS: We found no association between season of birth and schizophrenia, affective psychoses or neurotic depression. For schizophrenia, the pattern of risks by quarter reflects northern hemisphere trends. Results by month are difficult to interpret due to large fluctuations in the data. While age-incidence effects had no impact on the distribution of risk, we found an artefactual increase in January births due to routine imputation of missing birth dates. CONCLUSION: Adjusting for artefacts in the data produced a pattern analogous to northern hemisphere trends. PMID- 11473509 TI - Birth order and risk for schizophrenia: a 31-year follow-up of the Northern Finland 1966 Birth Cohort. AB - OBJECTIVE: The Northern Finland 1966 Birth Cohort was studied in order to investigate the association between birth order and schizophrenia. METHOD: Four categories of birth order status (first-born, last-born, only child and other status) were formed and linked to data on psychiatric morbidity. Effects were adjusted for wantedness of pregnancy, perinatal complications, maternal age at delivery, family type and number of siblings. RESULTS: The risk for schizophrenia was elevated among male first-borns (ratio 1.5; 95% CI 1.0-2.2) and female last borns (ratio 1.3; 95% CI 0.9-1.9). The risk was lower than expected among male last-borns (ratio 0.7; 95% CI 0.5-0.9) and females belonging to other status (ratio 0.6; 95% CI 0.3-0.9). CONCLUSION: These results suggest that specific birth order status is an independent risk factor for schizophrenia. Theoretical explanations may arise from biological factors unidentified here and/or psychological stressors linked with these positions. PMID- 11473510 TI - Rediscovering chronic mania. AB - OBJECTIVE: This paper presents a case of chronic mania and reviews the diagnosis with respect to its definition and utility. METHOD: A case of chronic mania is described. RESULTS: Unremitting mania poses a diagnostic and management challenge. CONCLUSION: Persistent mania of this duration is rare, but a diagnosis of chronic mania should not be rejected on the basis of chronicity. PMID- 11473527 TI - Anti-remodelling drugs for the treatment of asthma: requirement for animal models of airway wall remodelling. AB - 1. Airway wall remodelling (AWR), the structural change induced by acute and chronic inflammation in the airways, may be one of the most significant and difficult to reverse components of progressive asthma. 2. The mechanisms underlying the development of AWR are not known. Studies of only the most superficial wall structures of large airways can be conducted in living humans because of the degree of invasiveness required to measure airway structural changes. These studies reveal that currently available agents do not fully prevent or reverse AWR. Thus, animal models of asthma pathology may be used to assess the contribution of particular mediators and cells to the development of remodelling and may also prove to be useful in the initial screening of potential anti-remodelling agents. 3. Airway hyperresponsiveness and AWR stimulated by chronic antigen challenge in previously disease-free animals is the most popular of the currently used models of remodelling. Other animal models include the use of specially bred strains with intrinsic airway hyperresponsiveness or animals that have a naturally occurring asthma-like disease, such as cats with feline asthma or horses with heaves. The further development of animal models of AWR will facilitate the development of novel anti-asthma therapies. PMID- 11473528 TI - Hypervolaemia improves global and local function and efficiency in postischaemic myocardium. AB - 1. In the present study, we investigated the effects of blood volume on postischaemic function and efficiency. In 14 anaesthetized dogs, following recovery from a period of 15 min occlusion of the left anterior descending coronary artery, the effects of hypervolaemia (HYPER; 15% increased volume produced by fast infusion of Hespan; B Braun Medical, Irvine, CA, USA), normovolaemia (NORMO) and hypovolaemia (HYPO) were studied. 2. Although myocardial O2 consumption was not significantly increased by volume (6.37+/-0.94 vs 6.89+/-1.1 mL/min per 100 g for HYPO and HYPER, respectively), local work of the stunned myocardium was markedly elevated (8.8+/-1.7 vs 22.5+/-3.5 g.mm/ beat, for HYPO and HYPER, respectively; P < 0.05). External work of the heart was also significantly improved (71.8+/-12.7 vs 139.5+/-16.2 mmHg.L/min for HYPO and HYPER, respectively). These data indicate markedly improved efficiency produced by volume, because work was increased with no change in myocardial O2 consumption. 3. Local dysfunction was characterized by several parameters, including systolic bulge, end-diastolic length, delay to onset of shortening, end shortening time delay (EST) and tail work ratio. Hypervolaemia reduced EST compared with hypovolaemia (98.6+/-18.3 vs 110.7+/-14.9 msec, respectively; P < 0.05) and improved tail work ratio (28.0+/-7.0 vs 36.0+/-7.0%, respectively; P < 0.05), with no effects on systolic bulge, end-diastolic length and delay to onset of shortening. 4. Thus, even in the postischaemic myocardium, increasing work by volume is energetically efficient and is accompanied by partial improvement of local dysfunction. PMID- 11473529 TI - Effect of epomediol on ethinyloestradiol-induced changes in bile acid and cholesterol metabolism in rats. AB - 1. Epomediol is a terpenoid compound that has been reported to stimulate bile acid synthesis and to reverse 17alpha- ethinyloestradiol-induced cholestasis. The aim of the present study was to investigate the contribution of changes in bile acid and cholesterol metabolism to the protective effects of epomediol in ethinyloestradiol-treated rats. Animals received epomediol for 5 days at 100 mg/kg daily, i.p., ethinyloestradiol for 5 days at 5 mg/kg, s.c., or a combination of both drugs. 2. When compared with control animals, epomediol treatment resulted in a significant increase in bile flow (+42%) and in the secretion of bile acids (+74%) and cholesterol (+42%). Ethinyloestradiol administration caused a significant decrease in bile flow (-43%), bile acid secretion (-37%) and cholesterol secretion (-45%). Bile flow, bile acid secretion and cholesterol secretion were significantly increased in animals receiving ethinyloestradiol plus epomediol compared with ethinyloestradiol-treated rats (+13, +29 and +31%, respectively). 3. Both cholesterol 7alpha-hydroxylase and hydroxy-3- methylglutaryl coenzyme A reductase activities were significantly increased in epomediol-treated rats (+30 and +96%, respectively). Cholesterol 7alpha-hydroxylase activity was significantly reduced by ethinyloestradiol (-22%) and did not differ from control values in animals receiving epomediol plus ethinyloestradiol. Levels of cholesterol 7alpha-hydroxylase mRNA were elevated (+41%) by epomediol, but were not significantly modified by ethinyloestradiol or ethinyloestradiol plus epomediol. 4. It is concluded that epomediol enhances bile acid secretion by increasing the expression of cholesterol 7alpha-hydroxylase. Changes in bile acid metabolism contribute to the effects of epomediol in rats with ethinyloestradiol-induced cholestasis. PMID- 11473530 TI - Effects of adenosine administration on spike-wave discharge frequency in genetically epileptic rats. AB - 1. In the present study, the effects of the administration of adenosine on absence seizures were investigated in the Wistar Albino Glaxo/Rijswijk (WAG/Rij) strain of rats, which are an adequate model for human absence epilepsy. 2. After baseline electroencephalogram (EEG) recording, adenosine was injected intraperitoneally (i.p.) at doses of 6, 30 and 120 mg/kg and EEG recordings were continued for 1 h postinjection. In addition, to enable evaluation of the contribution of peripheral factors to the occurrence of spike-wave discharges (SWD) after adenosine injection, arterial blood pressure and rectal temperature were also recorded. 3. Injection of adenosine induced a dose-dependent increase in the appearance of SWD. The number and total duration of SWD were found to be significantly increased after 30 and 120 mg/kg adenosine (P < 0.05). Adenosine also induced a fall in both blood pressure and body temperature in all experimental groups. 4. These results show that peripheral administration of adenosine promotes absence seizures. It is likely that the lowering of blood pressure and body temperature and the activation of sensory afferents after adenosine injection may contribute to the facilitation of SWD observed in the present study. PMID- 11473531 TI - Role of endothelin-1 receptors in healthy anaesthetized rabbits. AB - 1. Many diseases are associated with elevated endothelin (ET)-1 plasma concentrations. In order to understand the consequence of this elevation, in the present study the effects of exogenous ET-1 on the entire organsim were investigated, in particular with respect to the role of ETA and ETB receptors in the cardiovascular system. In open-chest rabbits, left ventricular (LV) pressure (LVPmax, LVPed), dP/dtmax and dP/dtmin were recorded in ejecting and isovolumically beating hearts to determine cardiac function. In addition, heart rate (HR), aortic pressure (AoP) and aortic flow (AoF) were measured. Total peripheral resistance (TPR) was calculated from mean AoP and AoF. 2. In the first series of experiments (n = 11), ET-1 (0.5 nmol/kg; bolus) produced a non significant reduction in HR. Systolic function, in terms of AoF, LVPmax and dP/dtmax, was improved; for example, LVPmax was increased significantly (69 +/- 10 vs 106 +/- 20 mmHg for control and ET-1, respectively; P < 0.05). Similarly, early relaxation (dP/dtmin) was improved. In parallel, TPR rose significantly (0.25+/-0.07 vs 0.35+/-0.1 mmHg/min per mL for control and ET-1, respectively; P < 0.05). Isovolumic measurements showed corresponding responses. 3. In the second series of experiments (n = 7), animals were pretreated with an ETA receptor antagonist (330 nmol/min per kg FR 139317). After ETA receptor blockade, the administration of ET-1 had no significant effect on cardiac function or vasomotion. 4. In the third series of experiments (n = 6), animals were pretreated with an ETB receptor antagonist (10 nmol/min per kg BQ 788). In this series of experiments, the effects of ET-1 on cardiac function and vasomotion were the same as in the first series of experiments, except for the effect on HR, which decreased by 35% after ET-1. 5. In our experimental model, exogenous ET-1 exerted a clear-cut positive inotropic effect, together with the anticipated peripheral vasoconstriction via ETA receptors. PMID- 11473532 TI - Effects of dopamine d2 receptor agonists in a pituitary transplantation-induced hyperprolactinaemia/anovulation model in rats. AB - 1. In the present study, we investigated the effects of hyperprolactinaemia, induced by transplantation of anterior pituitary glands under the kidney capsule in female rats, on the relationship between serum and pituitary concentrations of the gonadotropins and on the oestrous cycle. 2. Rats with pituitary transplants showed increased serum prolactin concentrations and decreased serum concentrations of gonadotropins and increased pituitary concentrations of gonadotropins. Moreover, these rats showed persistent dioestrous and anovulation from 3 to 6 days after transplantation. 3. A single oral administration of cabergoline (at doses between 0.001 and 0.1 mg/kg) dose-dependently inhibited the elevated serum prolactin concentrations in hyperprolactinaemic rats. At 0.1 mg/kg, cabergoline induced a continuous reduction in serum prolactin concentrations for 5 days after administration. Terguride (0.1 mg/kg) and bromocriptine (10 mg/kg) also reduced serum prolactin concentrations at 1 and 3 days after administration. All three dopamine D2 receptor agonists increased serum gonadotropin concentrations and ovarian weight at 3 days after administration. 4. In rats exhibiting anovulation, a single oral administration of any one of the three dopamine D2 receptor agonists dose-dependently restored ovulation and a normal oestrous cycle appeared. Oral administration of cabergoline (0.03 mg/kg) or terguride (0.1 mg/kg) restored ovarian function and abolished the anovulation following a reduction in serum prolactin concentrations. However, bromocriptine (10 mg/kg) did not completely abolish anovulation. Following administration of terguride (0.3 mg/kg) or bromocriptine (30 mg/kg), only one normal oestrous cycle appeared; however, following cabergoline (0.1 mg/kg), two normal oestrous cycles appeared. 5. These results suggest that cabergoline has a potent and long-lasting action as a dopamine D2 receptor agonist and, thus, should be a useful drug for the treatment of galactorrhoea and hyperprolactinaemic amenorrhoea and/or anovulation in humans. PMID- 11473533 TI - Exacerbatory mechanism responsible for water immersion stress-induced gastric lesions in aged rats compared with young rats. AB - 1. The present study was designed to investigate whether or not ageing affects the development of water immersion stress-induced gastric lesions in rats. Effects of cetraxate, an anti-ulcer drug, were also examined. 2. Gastric lesions were induced by 6 h water immersion stress in rats. Gastric mucosal blood flow was determined by the hydrogen gas clearance technique and nitric oxide synthase (NOS) activity was measured enzymatically. 3. Early development of gastric lesions was observed in aged rats and exacerbation of gastric lesions was also found. Lowering of gastric mucosal blood flow and reduced NOS activity were observed in aged rats. 4. Cetraxate mitigated the development of gastric lesions in young rats and also increased gastric mucosal blood flow and NOS activity. However, these favourable effects were diminished in aged rats. 5. Decreased NOS activity may be an important exacerbatory factor to the development of gastric lesions in aged rats. 6. Effects of cetraxate differed between young rats and aged rats. 7. These results may explain the refractoriness and drug resistance in gastric ulcers encountered by elderly individuals. PMID- 11473534 TI - General practitioner participation in the second Australian National Blood Pressure Study (ANBP2). AB - 1. The second Australian National Blood Pressure Study (ANBP2) is an outcome trial of the treatment of hypertension in the elderly conducted entirely in general practices across Australia. Prior to ANBP2, no study of this size and nature had been undertaken in Australian general practice and the response of General Practitioners (GPs) to becoming involved in long-term cardiovascular research was unknown. 2. Academic departments and Divisions of General Practice were approached to support the project. General Practitioners were approached by letter of invitation and contacted by a regional medical coordinator (RMC) either at a face-to-face meeting or by telephone. 3. At the close of recruitment to ANBP2, 1938 GPs from 950 practices had registered as investigators. Sixty-two Divisions of General Practice were approached to support the study in five mainland Australian states with 39 (63%) participating, although participation by state was highly variable (range: 18-100%). Thirty divisional or promotional dinner meetings were held, with 56% (368/658) of those attending registering as investigators. Of the 8098 GPs sent a letter of invitation to participate in the study, 1357 (17%) expressed interest and eventually enrolled as investigators, ranging from 8% in Queensland to 28% in New South Wales. Ninety-six per cent of GPs who had a personal face-to-face contact (696/724) with the RMC registered in the study. 4. The GP recruitment phase of ANBP2 has been successfully completed. Peer-to-peer recruitment was the most successful strategy; however, success varied between states. General Practitioner recruitment to long-term clinical trials appears to be successful with a multifactorial approach focusing on peer to-peer recruitment. PMID- 11473535 TI - "Current" advances in mechanically skinned skeletal muscle fibres. AB - 1. In skeletal muscle, excitation-contraction (E-C) coupling describes a cascade of cellular events initiated by an action potential (AP) at the surface membrane that ultimately results in muscle contraction. Being able to specifically manipulate the many processes that constitute E-C coupling, as well as the many factors that modulate these processes, has proven challenging. 2. One of the simplest methods of gaining access to the intracellular environment of the muscle fibre is to physically remove (mechanically skin) the surface membrane. In doing so, the myoplasmic environment is opened to external manipulation. 3. Surprisingly, even though the surface membrane is absent, it is still possible to activate both twitch and tetanic force responses in a mechanically skinned muscle fibre by generating an AP in the transverse tubular system. This proves that all the key steps in E-C coupling are retained in this preparation. 4. By using this technique, it is now possible to easily manipulate the myoplasmic environment and observe how altering individual factors affects the normal E-C coupling sequence. The effect of important factors, such as the redox state of the cell, parvalbumin and the sarcoplasmic reticulum Ca2+-ATPase, on twitch and tetanic force can now be specifically investigated independent of other factors. PMID- 11473536 TI - The power of single channel recording and analysis: its application to ryanodine receptors in lipid bilayers. AB - 1. Since the inception of the patch-clamp technique, single-channel recording has made an enormous impact on our understanding of ion channel function and its role in membrane transport and cell physiology. 2. However, the impact of single channel recording methods on our understanding of intracellular Ca2+ regulation by internal stores is not as broadly recognized. There are several possible reasons for this. 3. First, ion channels in the membranes of intracellular organelles are not directly accessible to patch pipettes, requiring other methods that are not as widely known as the patch-clamp techniques. 4. Second, bulk assays for channel activity have proved successful in advancing our knowledge of Ca2+ handling by intracellular stores. These assays include Ca2+ imaging, ryanodine binding assays and measurements of muscle tension and Ca2+ release and uptake by vesicles that have been isolated from internal stores. 5. The present review describes methods used for single- channel recording and analysis, as applied to the calcium release channels in striated muscle, and details some of the unique contributions that single-channel recording and analysis have made to our current understanding of the release of Ca2+ from the internal stores of muscle. 6. With this in mind, the review focuses on three aspects of channel function and shows how single-channel investigations have led to an improved understanding of physiological processes in muscle. 7. Finally, the review describes some of the latest improvements in membrane technology that will underpin future advances in single-channel recording. PMID- 11473537 TI - Gene transfer: manipulating and monitoring function in cells and tissues. AB - 1. The ectopic expression of genes has proven to be an extremely valuable tool for biologists. The most widely used systems involve electrically or chemically mediated transfer of genes to immortalized cell lines and, at the other end of the spectrum, transgenic animal models. As would be expected, there are compromises to be made when using either of these broad approaches. Immortalized cell lines have limited "physiological relevance" and transgenic approaches are costly and out of the reach of many laboratories. There is also significant time required for the de novo generation of a transgenic animal. 2. As a viable alternative to these approaches, we describe the use of recombinant adenovirus and Sindbis virus to deliver genes to cells and tissues. 3. We exemplify this approach with studies from our laboratories: (i) an investigation of Ca2+ handling deficits in cardiac myocytes of hypertrophied hearts using infection with recombinant adenovirus encoding either green fluorescent protein (GFP) or the sarcoplasmic/endoplasmic reticulum calcium-ATPase (Serca2a); (ii) a study of the mechanism of macrophage/microglial migration by infection of embryonic phagocytes with a GFP-encoding virus and coculture with brain slices to then track the movement of labelled cells; and (iii) we are also exploiting the natural tropism of the Sindbis virus to label neurons in hippocampal brain slices in culture to resolve high-resolution structure and to map neuronal connectivity. 4. Further development of these approaches should open new avenues of investigation for the study of physiology in a range of cells and tissues. PMID- 11473538 TI - Hybrid skeletal muscle fibres: a rare or common phenomenon? AB - 1. The main aim of the present review is to raise awareness of the molecular complexity of single skeletal muscle fibres from "normal" and "transforming" muscles, in recognition of the many types of hybrids that have been observed in vertebrate skeletal muscle. The data used to illustrate various points made in the review were taken from studies on mammalian (mostly rat) and amphibian muscles. 2. The review provides a brief overview of the pattern and extent of molecular heterogeneity in hybrid muscle fibres and of the methodological problems encountered when attempting to identify and characterize such fibres. Particular attention is given to four types of skeletal muscle hybrids: (i) myosin heavy chain (MHC) hybrids; (ii) mismatched MHC-myosin light chains (MLC) hybrids; (iii) mismatched MHC-regulatory protein hybrids; and (iv) hybrids containing mismatched MHC-sarcoplasmic reticulum protein isoforms. 3. Some of the current ideas regarding the functional significance, origin and cognitive value of hybrid fibres are examined critically. PMID- 11473539 TI - Does Ca2+ release from the sarcoplasmic reticulum influence heart rate? AB - 1. The present review summarizes the evidence that Ca2+ release from the sarcoplasmic reticulum (SR) is an important contributor to the systolic rise in [Ca2+]i (the Ca2+ transient) and influences the pacemaker firing rate. 2. We believe that the mechanism whereby [Ca2+]i influences firing rate is through the dependence of the Na+-Ca2+ exchanger on [Ca2+]i. 3. Extrusion of Ca2+ by the electrogenic Na+-Ca2+ exchanger produces an inward current that contributes to the pacemaker currents. Confocal images of Ca2+ indicate the distribution of [Ca2+]i and Ca2+ sparks add to the evidence that Ca2+ release from SR is involved in pacemaker activity. 4. The normal pathway for increased heart rate is sympathetic activation; we discuss the evidence that part of the chronotropic effect of beta-adrenoceptor stimulation is through the modulation of SR Ca2+ release. 5. These studies show that Ca2+ handling by the pacemaker cells makes an important contribution to the regulation of pacemaker activity. PMID- 11473540 TI - Of mice, frogs and flies: generation of membrane asymmetries in early development. AB - Embryonic development begins with cleavage of the fertilized egg. Cleavage comprises two major processes: cytokinesis and formation of a polarized epithelial cell layer. The focus of this review is comparison of the generation of membrane polarity during embryonic cleavage in three different developmental model systems. In mammalian embryos, as exemplified by analysis of the mouse, generation of distinct membrane domains is uncoupled from cleavage divisions and is initiated in a specific developmental phase, called compaction. In Xenopus laevis embryos, generation of polarized blastomeres occurs simultaneously with cytokinesis. The origin of specific membrane domains of X. laevis polar blastomeres, however, can be traced back to oogenesis. Finally, in Drosophila melanogaster, generation of polarized cells occurs at cellularization. The relevance of cell adhesion, cell junctions and cytocortical scaffolds will be discussed for each of the model systems. Despite enormous morphologic differences, the three models share many common features; in particular, many important molecular interactions are conserved. PMID- 11473541 TI - Fertilization and the first cleavage mitosis in insects. AB - Fertilization in animals is now considered to be of the "sea urchin type"; that is, haploid male and female pronuclei completely fuse shortly after sperm entry into the egg, followed by the formation of a mitotic spindle to allow cleavage mitoses to proceed. However, two other patterns of fertilization and early embryonic mitosis in some animal species are known: an Ascaris type and a gonomeric type. The gonomeric type of fertilization in insects and other arthropods is not well known and is quite different from the sea urchin and Ascaris types. In the present article, the author examines the peculiar gonomeric fertilization, using mainly the silkworm as an example. PMID- 11473542 TI - Comparative study of sequential expression of the organizer-related genes in normal Cynops pyrrhogaster embryos and mesodermalized ectoderm. AB - An artificially mesodermalized ectoderm (mE) of early Cynops pyrrhogaster gastrula acquires the organizer property; the mE is able to induce the secondary axis. The expression of organizer-related genes was investigated during the mesodermalizing process of the mE. The expression of C. pyrrhogaster organizer related genes, such as bra, gsc, lim-1, chd and noggin, were analyzed. Cynops pyrrhogaster shh expression was also investigated. The organizer-related genes were activated by 12 h after the mesoderm-inducing stimulus. It was noted that there was a temporal gap in the expression of each gene. The expression of bra and gsc seemed to be more quickly activated during the mesodermalizing process. While expression of lim-1 and noggin was activated later than that of bra and gsc, lim-1 expression was earlier than chd and noggin expression. Shh expression was activated later than lim-1/noggin. The present study suggests the possibility that the bra/gsc, lim-1, chd, noggin and shh genes are expressed one by one in that order during the mesodermalizing of the presumptive ectoderm. It also indicates that the sequence is not always consistent with that of the whole embryo during normal embryogenesis. The meaning of the discrepancy will be discussed in connection with the cascade of certain genes expressed during the mesodermalizing process. PMID- 11473543 TI - Highly efficient transfection system for functional gene analysis in adult amphibian lens regeneration. AB - The analysis of newt lens regeneration has been an important subject in developmental biology. Recently, it has been reported that the genes involved in the normal eye development are also expressed in the regenerative process of lens regeneration in the adult newt. However, functional analysis of these genes has not been possible, because there is no system to introduce genes efficiently into the cells involved in the regeneration. In the present study, lipofection was used as the method for gene transfer in cultured pigmented iris cells that can transdifferentiate into lens cells in newt lens regeneration. Positive expression of a reporter gene was obtained in more than 70% of cells. In addition, the aggregate derived from gene-transfected cells maintained its expression at a high level for a long time within the host tissue. To verify the effectiveness of this model system with a reporter gene in lens regeneration, Pax6, which is suggested to be involved in normal eye development and lens regeneration, was transfected. Ectopic expression of lens-specific crystallins was obtained in cells that show no such activity in normal lens regeneration. These results made it possible for the first time to analyze the molecular mechanism of lens regeneration in the adult newt. PMID- 11473544 TI - Developmental fates of the first four blastomeres of the chaetognath Paraspadella gotoi: relationship to protostomes. AB - Experimental analysis of the development of chaetognaths is virtually lacking. To elucidate developmental fates, single blastomeres of the 2-cell and 4-cell embryos of Paraspadella gotoi were injected with a lineage-tracing dye (Fluoro Ruby or DiI). The distribution of the labels was observed in the hatchlings. In a previous study, embryos were injected at the 2-cell stage with Fluoro-Ruby and two sets of complementary labeling patterns (DL and VR, and DR and VL) were found. The same results were obtained when DiI was used as a tracer dye. The 4 cell embryo consists of the animal and vegetal cross-furrow cells in a tetrahedral arrangement and one of the vegetal cross-furrow cells typically contains the germ plasm. When single cells were injected at the 4-cell stage, four labeling patterns were observed (D, V, L and R). These four patterns represent subsets of the four patterns observed in the hatchling injected at the 2-cell stage. The V pattern is probably generated from the blastomere containing the germ plasm. It was found that the positions of the blastomeres at the 4-cell stage corresponded to the future body axes, similar to classic spiralians and modified spiralians such as crustaceans. Furthermore, it was confirmed that second cleavage occurs in a leiotropic fashion, which is seen in the second cleavage of the classic spiralians. Chaetognaths may have some similarities to protostomes in their developmental program. PMID- 11473545 TI - Activation of the maternally preset program of apoptosis by microinjection of 5 aza-2'-deoxycytidine and 5-methyl-2'-deoxycytidine-5'-triphosphate in Xenopus laevis embryos. AB - The present study examines the effects on embryogenesis of microinjecting Xenopus laevis fertilized eggs with 5-aza-2'-deoxycytidine (5-Aza-CdR), which induces hypomethylation of DNA, and 5-methyl-2'- deoxycytidine-5'-triphosphate (5-methyl dCTP), which induces hypermethylation of DNA. Embryos injected with either one of these analogs cleaved normally until the mid-blastula stage, but underwent massive cell dissociation and stopped development at the early gastrula stage. Dissociated cells that appeared here were positive by terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick end-labeling and contained fragmented nuclei with condensed chromatin. The DNA from these cells formed a "ladder" on electrophoresis. Furthermore, the induction of cell dissociation by 5-Aza-CdR and 5-methyl-dCTP was postponed by 2 3 h by co-injection of Bcl-2 mRNA and the normal metabolite (CdR and dCTP, respectively). Using a specific antibody against 5-methyl-cytosine, we confirmed that 5-Aza-CdR induces hypomethylation, whereas 5-methyl-dCTP induces hypermethylation in X. laevis embryos before the onset of cell dissociation. Incorporation of radioactive precursors revealed that synthesis of DNA, and also RNA, is inhibited significantly in both 5-Aza-CdR-injected and 5-methyl-dCTP injected embryos. These results show that 5-Aza-CdR and 5-methyl-dCTP are incorporated into DNA and induce apoptosis, probably through alteration of DNA methylation coupled with inhibition of DNA replication and/or transcription. PMID- 11473546 TI - Possible roles of beta-catenin in evagination of the optic primordium in rat embryos. AB - The roles of beta-catenin in evagination of the optic primordium in rat embryos were studied using immunostaining. High levels of beta-catenin appeared transiently in the evaginating optic primordium. Evagination of the optic primordium was suppressed in embryos treated with LiCl. In deficient optic vesicles of these embryos, accumulation of beta-catenin was decreased. Deficient optic vesicles also showed suppression of cyclin D1 accumulation and bromodeoxyuridine incorporation, no break in the deposition of laminin and type IV collagen at the basement membrane (BM) and prevention of the change in distribution of microtubules and microfilaments. These results suggest that beta catenin regulates cell proliferation, breakdown of BM and changes in cell shape in the evaginating optic primordium to cause optic vesicle formation. PMID- 11473547 TI - Involvement of a urethane-sensitive system in timing the onset of gastrulation in Xenopus laevis embryos. AB - This paper describes success in delaying the onset of gastrulation in Xenopus laevis embryos without damage to their subsequent development by temporarily arresting cleavage with urethane. Exposure of X. laevis embryos to 150 mM urethane before gastrulation resulted in cleavage arrest and its removal led to cleavage resumption. During cleavage arrest, cyclic activities including nuclear replication and the M-phase-promoting factor cycle continued, although their duration was lengthened to nearly 1.8-fold that of the controls. Because of a 30 min time lag from removal of urethane to resumption of cleavage, as well as the retardation of cyclic activities during cleavage arrest, the development of embryos after a 60-min exposure to urethane lagged two cell cycles behind that of control embryos. Here, the two cell cycle delay is equivalent to 50 min at 22-23 degrees C. The start of gastrulation in exposed embryos was accordingly delayed about 50 min, although the delay in mid-blastula transition was as little as 20 25 min. Consistent results were obtained in embryos exposed to urethane for 90 or 120 min and those exposed to procaine or NH4Cl for 60 min. Although these results imply that delay in the start of gastrulation in exposed embryos is ascribed simply to delay in their development raised by cleavage arrest, at the same time they suggest that the onset of gastrulation is timed by systems sensitive to urethane, procaine and NH4Cl in X. laevis embryos. PMID- 11473548 TI - Inhibitors of procollagen C-terminal proteinase block gastrulation and spicule elongation in the sea urchin embryo. AB - In the sea urchin embryo, inhibition of collagen processing and deposition affects both gastrulation and embryonic skeleton (spicule) formation. It has been found that cell-free extracts of gastrula-stage embryos of Strongylocentrotus purpuratus contain a procollagen C-terminal proteinase (PCP) activity. A rationally designed non-peptidic organic hydroxamate, which is a potent and specific inhibitor of human recombinant PCP (FG-HL1), inhibited both the sea urchin PCP as well as purified chick embryo tendon PCP. In the sea urchin embryo, FG-HL1 inhibited gastrulation and blocked spicule elongation, but not spicule nucleation. A related compound with a terminal carboxylate rather than a hydroxamate (FG-HL2) did not inhibit either chick PCP or sea urchin PCP activity in a procollagen-cleavage assay. However, FG-HL2 did block spicule elongation without affecting spicule nucleation or gastrulation. Neither compound was toxic, because their effects were reversible on removal. It was shown that the inhibition of gastrulation and spicule elongation were independent of tissue specification events, because both the endoderm specific marker Endo1 and the primary mesenchyme cell specific marker SM50 were expressed in embryos treated with FG-HL1 and FG-HL2. These results suggest that disruption of the fibrillar collagen deposition in the blastocoele blocks the cell movements of gastrulation and may disrupt the positional information contained within the extracellular matrix, which is necessary for spicule formation. PMID- 11473549 TI - Morphologic and functional development of whole human fetal stomachs grafted into nude mice. AB - To study in vivo the cellular differentiation and secretion of human developing fetal stomach, ethically and technically impossible to perform in utero, 256 fetal stomachs were xenografted. Human stomachs from 6- to 10-week-old fetuses were grafted for 1-273 days into nude mice. Biopsies for immunohistochemistry, hybridization and electron microscopy were taken and a catheter introduced into the human stomach. Macroscopic growth was fast and cells in S phase were numerous during the first 9 weeks, then the stomach size was stable and the gastric mucosa, of adult type, remained normal. In situ hybridization detected only a minute mouse mesenchymal chimerism in the graft. Chromogranin A, intrinsic factor and H+/K+ adenosine triphosphatase were immunohistolocally detected in epithelial cells 20 days after grafting, gastrin was detected after 30 days and pepsinogen after 60 days. The pH in gastric juice, which was at 8.0 +/- 0.1 from days 10-25, dropped from 4.39 +/- 1.80 at 30 days to 1.58 +/- 0.29 at 90 days. Intrinsic factor was stable and pepsin ranged from 6.8 +/- 7.8 to 134 +/- 51 units at 90 days. The differentiation of the epithelial cells in xenografts was very accelerated in comparison to that in utero. PMID- 11473550 TI - Centriole behavior during meiosis in oocytes of the sea urchin Hemicentrotus pulcherrimus. AB - Ultrastructural changes in the maturing oocyte of the sea urchin Hemicentrotus pulcherrimus were observed, with special reference to the behavior of centrioles and chromosomes, using oocytes that had spontaneously started the maturation division process in vitro after dissection from ovaries. The proportion of oocytes entering the maturation process differed from batch to batch. In those eggs that accomplished the maturation division, it took ~4.5-5 h from the beginning of germinal vesicle breakdown to the formation of a second polar body. Serial sections revealed that a young oocyte before germinal vesicle breakdown had a pair of centrioles with procentrioles, located between the presumed animal pole and the germinal vesicle and accompanied by amorphous aggregates of moderately dense material and dense granules (granular aggregate). Just before germinal vesicle breakdown, a pair of fully grown centrioles located in the granular aggregate, which is present until this stage and then disappears, had already separated from another pair of centrioles. In meiosis I, each division pole had two centrioles, whereas in meiosis II each had only one. The two centrioles in the secondary oocyte separated into single units and formed the mitotic figure of meiosis II. The first polar body had two centrioles and the second had only one. The two centrioles in the first polar body did not form the mitotic figure nor did they separate at the time of meiosis II. These results indicate that, in sea urchins, duplication of the centrioles does not occur during the two successive meiotic divisions and the egg inherits only one centriole from the primary oocyte, confirming the results previously reported for starfish oocytes. PMID- 11473551 TI - Measurement of the intracellular pH threshold for sperm aster formation in sea urchin eggs. AB - In the fertilization of sea urchin eggs, intracellular [Ca2+] (Cai) increases transiently and intracellular pH (pHi) elevates accordingly. Unlinking these two activating factors experimentally, the requirement of the increase in pHi for sperm aster formation in the sea urchin, Clypeaster japonicus, was investigated. When the eggs were injected with an EGTA or BAPTA solution, they incorporated sperm but did not organize the sperm aster. Using these sperm-incorporated eggs under the condition that an increase in Cai was blocked, pHi was regulated by two methods: (i) perfusing ammonium acetate-containing seawater; and (ii) injecting pH buffer solutions of various pH values. By either of the two methods, the sperm aster formed at pHi 7.0 or more and functioned in female pronuclear migration when the sperm aster reached the female pronucleus. Hence, the step of the transient increase in Cai at fertilization can be bypassed. In contrast, a pHi increase is indispensably required for sperm aster formation in sea urchin eggs. Moreover, under the condition that there was the transient increase in Cai, the threshold pHi value for sperm aster formation was pHi 7.0 or more. Consequently, whether a Cai increase on fertilization occurs or not, the threshold pHi value for sperm aster formation is constant in sea urchin eggs. PMID- 11473552 TI - Evolution of larval form in the sea star genus Patiriella: conservation and change in the larval nervous system. AB - The organization of the peptidergic system in the larvae of Patiriella species with divergent ontogenies was compared to determine which aspects of neurogenesis are conserved and which are altered in the evolution of development in these sea stars. P. regularis has ancestral-type feeding bipinnaria and brachiolaria larvae and the organization of the nervous system, in association with feeding structures, paralleled the bilateral larval body plan. P. calcar and P. exigua have non-feeding planktonic and benthic brachiolariae, respectively, and there was no trace of the neuronal architecture involved with feeding. The nervous system in the attachment stage brachiolaria was similar in all three species and neuronal organization reflected larval symmetry. Delayed expression of peptidergic lineages to the brachiolaria stage in the lecithotrophs indicates heterochronic change in the timing of neurogenesis or deletion of the ancestral early neurogenic program. The bipinnarial program is suggested to be a developmental module autonomous from the brachiolar one. With a divergence time of less than 10 Ma, the evolution of development in Patiriella has resulted in extensive reduction in the complexity of the larval nervous system in parallel with simplification in larval form. There is, however, strong conservation in the morphology and neuronal architecture of structures involved with settlement. PMID- 11473553 TI - Transfusion strategy. PMID- 11473554 TI - Transfusion strategy. AB - As is often the case in medicine, the risk to benefit ratio of blood transfusion requires careful examination. The decision to transfuse a patient cannot be based only on the haemoglobin or the haematocrit concentration. In a given clinical situation, tolerance to anaemia will depend on the ability of the patient to recruit his compensatory mechanisms in accordance with his metabolic demand. Maintenance of circulating volume is of paramount importance. In the absence of a universal transfusion trigger, decision to transfuse a patient with an haemoglobin concentration between 7.0 and 10.0 g dL-1 depends primarily on clinical judgement. With the exception of emergency situations, blood transfusion will be realized on a unit-by-unit basis, and the efficacy of treatment should be evaluated after each transfused unit. PMID- 11473555 TI - Low-flow anaesthesia with desflurane: kinetics during clinical procedures. AB - BACKGROUND AND OBJECTIVE: Low-flow anaesthesia is economical and less polluting. The purpose of this study was to determine the inspired and end-tidal desflurane concentrations during anaesthesia with a vaporizer setting maintained at 5%, during low-flow anaesthesia after 120 min with fresh gas inflows of 1.0 and 2.0 L min-1. METHODS: The study was both prospective and randomized, including 56 patients (28 patients in each group) scheduled for elective surgery of an expected anaesthesia time of at least 120 min. Inspired and end-tidal concentrations of desflurane were measured during low-flow anaesthesia with fresh gas inflows of 1.0 and 2.0 L min-1. The vaporizer setting was fixed at 5% desflurane. RESULTS: The inspired and end-tidal concentrations of desflurane in the 1.0 L min-1 group after 120 min were 4.54% vs. 4.37% (P < 0.001). In the 2.0 L min-1 group, the inspired and end-tidal concentrations of desflurane were 4.76% vs. 4.58% (P < 0.001). The estimated end-tidal/inspired ratios at 120 min of anaesthesia were 0.96 in both groups. At a fresh gas flow of 1.0 L min-1, the end tidal concentration was 0.87 of the vaporizer setting. Increasing the fresh gas flow to 2.0 L min-1 increased the end-tidal value by 0.05. CONCLUSION: There is a significant difference between the inspired and end-tidal concentrations of desflurane when fresh gas inflows were 1.0 and 2.0 L min-1, but not for the ratio of inspired/end-tidal. PMID- 11473556 TI - Painful regional anaesthesia induces an immunological stress reaction: the model of retrobulbar anaesthesia. AB - BACKGROUND AND OBJECTIVE: Laboratory stress studies found that acute psychological stresses may elicit changes in leukocyte numbers similar to those occurring in physical stresses. Both types of stress evoke - mainly by release of catecholamines - leukocytosis resulting from a release of natural killer cells (NK-cells), of CD8+ T-cells, of monocytes and of neutrophils. However, there is little proof that laboratory stress models can be applied to daily clinical routines. As a likely inductor of an immunological stress response the setting of retrobulbar anaesthesia prior to intraocular surgery permits the study of a short term painful anaesthetic procedure under highly standardized conditions. This was examined in 16 female patients. METHODS: Counts of leukocyte subsets, serum cortisol and cardiovascular variables were measured 30 min and 1 min prior to retrobulbar anaesthesia as well as 2, 15 and 45 min afterwards. RESULTS: The setting of retrobulbar anaesthesia induced an increase in total leukocytes [+380 cells microL-1; P < 0.01 (means; significance level)] mainly due to rising counts of neutrophils (+241 cells microL-1, P < 0.01). Of all lymphocyte subpopulations, natural killer cells increased most markedly (+64 cells microL-1; P < 0.01). Furthermore, the retrobulbar block induced an increase in systolic arterial pressure (+15.2 mmHg; P < 0.01). CONCLUSION: These changes in immunological and cardiovascular variables are considered to be elements of a sympatho-adrenal stress reaction; catecholamines are considered to induce a demargination of leukocytes by binding to beta2-adrenoceptors and by modifying the avidity state of adhesion molecules. PMID- 11473557 TI - Morbidity in electroconvulsive therapy. AB - BACKGROUND AND OBJECTIVE: To assess retrospectively the complications and morbidity of electroconvulsive therapy. METHODS: Complications occurring in 75 patients during 612 electroconvulsive therapy procedures under propofol anaesthesia were reviewed by data analysis. RESULTS: At least one complication occurred in 51 patients (68%) during the course of their treatment. Among these complications, 12 were potentially life-threatening: one patient developed angina pectoris, another aspiration pneumopathy, there were two incidences of bronchospasm, three hypoxic episodes (SpO2 < 92% with FiO2=1) and five severe episodes of laryngospasm which caused hypoxia. Twenty-five patients (33%) were confused for more than 2 h after the electroconvulsive therapy. Confusion recurred in 10 patients (13%) after several sessions of electroconvulsive treatment. Six patients had a traumatic complication, with one requiring surgery. CONCLUSION: Our results, compared with other studies, suggest that electroconvulsive therapy is not a low-risk procedure, with a particularly high rate of respiratory complications that may have been previously overlooked. Therefore, ambulatory anaesthesia may not be appropriate on a regular basis for most of these patients. PMID- 11473558 TI - The influence of fentanyl vs. s-ketamine on intubating conditions during induction of anaesthesia with etomidate and rocuronium. AB - BACKGROUND AND OBJECTIVE: In the present study, we investigated the combination of etomidate and s-ketamine with regard to its suitability for modified rapid sequence induction using rocuronium for muscle relaxation. METHODS: In a prospective, randomized and double-blinded study, 90 patients were assigned to one of three groups for induction of anaesthesia in combination with etomidate (0.3 mg kg-1) and muscle relaxation with rocuronium (0.6 mg kg-1). The groups were as follows: (a) control, i.e. placebo; (b) fentanyl, fentanyl (1.5 microg kg 1); (c) ketamine, s-ketamine (0.5 mg kg-1). Tracheal-intubating conditions after 1 min were classified as excellent, good or poor. During the induction of anaesthesia, arterial pressure and heart rate were measured every 60 s. RESULTS: Intubating conditions were best using etomidate and s-ketamine (23 excellent, 7 good, 0 poor) compared with the control (8, 16, 6 respectively) and fentanyl groups (7, 21, 2 respectively) (P < 0.01). While heart rate and arterial pressure remained stable in the control and fentanyl groups during induction, both significantly increased in the ketamine group (P < 0.01). CONCLUSIONS: The combination of etomidate and s-ketamine for anaesthesia induction produces mostly excellent intubating conditions after 60 s using only 0.6 mg kg-1 of rocuronium. This combination of drugs may be a useful alternative, if succinylcholine needs to be avoided, for modified rapid-sequence induction. PMID- 11473559 TI - The effect of lidocaine on neutrophil respiratory burst during induction of general anaesthesia and tracheal intubation. AB - BACKGROUND AND OBJECTIVE: Respiratory burst is an essential component of the neutrophil's biocidal function. In vitro, sodium thiopental, isoflurane and lidocaine each inhibit neutrophil respiratory burst. The objectives of this study were (a) to determine the effect of a standard clinical induction/tracheal intubation sequence on neutrophil respiratory burst and (b) to determine the effect of intravenous lidocaine administration during induction of anaesthesia on neutrophil respiratory burst. METHODS: Twenty ASA I and II patients, aged 18-60 years, undergoing elective surgery were studied. After induction of anaesthesia [fentanyl (2 microg kg-1), thiopental (4-6 mg kg-1), isoflurane (end-tidal concentration 0.5-1.5%) in nitrous oxide (66%) and oxygen], patients randomly received either lidocaine 1.5 mg kg-1 (group L) or 0.9% saline (group S) prior to tracheal intubation. Neutrophil respiratory burst was measured immediately prior to induction of anaesthesia, immediately before and 1 and 5 min after lidocaine/saline. RESULTS: Neutrophil respiratory burst decreased significantly after induction of anaesthesia in both groups [87.4 +/- 8.2% (group L) and 88.5 +/- 13.4% (group S) of preinduction level (P < 0.01 both groups)]. After intravenous lidocaine (but not saline) administration, neutrophil respiratory burst returned towards preinduction levels, both before (97.1 +/- 23.6%) and after (94.4 +/- 16.6%) tracheal intubation. CONCLUSION: Induction of anaesthesia and tracheal intubation using thiopentone and isoflurane, inhibit neutrophil respiratory burst. This effect may be diminished by the administration of lidocaine. PMID- 11473560 TI - Laparoscopically assisted vaginal and abdominal hysterectomy: comparison of postoperative pain, fatigue and systemic response. A case-control study. AB - BACKGROUND AND OBJECTIVE: Laparoscopic and open surgery have been compared with conflicting results regarding their systemic responses. The sensitivity of biochemical markers that are used to discriminate between the stress responses to different types of surgery varies from study to study. We wanted to evaluate the stress response and the sensitivity of clinical and biochemical stress markers in patients undergoing laparoscopically assisted vaginal or abdominal hysterectomy. METHODS: We performed a case-control study with patients undergoing laparoscopically assisted vaginal hysterectomy (n=20) or abdominal hysterectomy (n=20). Pain scores were assessed at rest and during coughing, and active leg elevation and fatigue scores using a visual analogue scale. In 10 patients of each group, haematocrit, white cell count, C-reactive protein, glucose, cortisol, adrenocorticotrophic hormone, beta-endorphin immunoreactivity, interleukin-6 and urine excretion of epinephrine and norepinephrine were measured preoperatively and during the first 44 postoperative hours. RESULTS: The most sensitive symptoms and markers of the systemic response were pain scores during mobilization, fatigue scores, C-reactive protein and interleukin-6 (P < 0.01 in all comparisons). Pain scores at rest, and all other laboratory markers of the systemic response, did not discriminate between the two types of surgery. CONCLUSION: Follow-up of postoperative pain scores during mobilization and fatigue levels might be an easy tool for the evaluation of postoperative recovery. Using an identical anaesthetic technique, the neuroendocrine response was of the same magnitude after both types of surgery. PMID- 11473561 TI - Management and outcome of patients undergoing thoracic surgery in a regional chest medical centre. AB - BACKGROUND AND OBJECTIVE: The main objective of this study was to assess mortality and morbidity after thoracic surgery in a medical centre, without resident chest surgeons and anaesthesiologists, and to determine specific risk factors. METHODS: A prospective cohort study using a local database which includes patients' clinical characteristics, results of preoperative investigations, surgical and anaesthesia data and all postoperative complications was undertaken. Two hundred and seventy-three consecutive patients undergoing thoracic surgery from 1992 to 1999 were studied. The referral chest medical centre was without resident thoracic surgeons or anaesthesiologists; postoperative care was led by local chest physicians according to standardized protocols and in close collaboration with university-based surgeons and anaesthesiologists. RESULTS: The majority of patients had lung cancer (71%) and underwent resection of at least one lobe (62%). Thirty-day mortality rate was 2.2% and one or more complications occurred in 74 patients (27%). Three patients had to be transferred to a university hospital for further treatment. Univariate predictors of complications included age (> 70 years), history of smoking, body mass index, as well as the extent and duration of surgery. After multiple logistic regression analysis, smoking (current or past), prolonged surgery (>120 min) and major lung resection (pneumonectomy or bilobectomy) remained the only independent risk factors. CONCLUSIONS: Overall perioperative mortality and morbidity rates did not exceed those reported from large teaching hospitals. In selected patients, thoracic surgery can be safely performed in a specialized chest medical centre without on-site surgeons and anaesthesiologists. PMID- 11473562 TI - Fatal form of phaeochromocytoma presenting as acute pyelonephritis. AB - We report the case of a young man who presented with a clinical picture of acute pyelonephritis. Within 3 h of admission, the patient developed acute respiratory distress associated with tachycardia and shock, and he was transferred to the intensive care unit. Mechanical ventilation of the lungs and symptomatic treatment were started immediately. Abdominal ultrasound revealed the presence of an adrenal tumour with central necrosis indicating a probable phaeochromocytoma. There was no sign of pyelonephritis. Ventricular fibrillation followed by asystole occurred soon after admission. The suddenness of the patient's death did not allow time for further investigation and therapy. The severity of the clinical signs was probably related to a massive release of catecholamines because of necrosis of the tumour, which may have been worsened by the diagnostic procedures performed to investigate the clinical symptoms and signs of acute pyelonephritis. PMID- 11473563 TI - Vasovagal syncope and anaesthetic practice. AB - We surveyed anaesthetists working in North-West England and in North Wales concerning episodes of vasovagal syncope encountered in their practice. Eighty eight anaesthetists described 109 such events occurring in either patients or their relatives and the estimated frequency of syncope was 1 in 5000 anaesthetic episodes. The patient syncopal episodes were triggered by venous cannulation in 16 instances and regional or local techniques in 20 instances. Thirty-three of the 53 patients were in the upright position when syncope occurred. Thirty-nine of the 56 relatives were male partners of female patients and four of these partners suffered some morbidity. The results of the survey are consistent with our current knowledge of the pathophysiology of vasovagal syncope, which is summarized, and also highlight the common anaesthetic scenarios where fainting is most likely to occur. Given this information anaesthetists will be in a better position to avoid such potentially harmful episodes in the future. PMID- 11473564 TI - Effects of hydroxyethylstarch infusion on drug protein binding in critically ill patients. PMID- 11473565 TI - Seppo Yla-Herttuala receives the ESCI Award for Excellence in Clinical Science 2000. PMID- 11473566 TI - Gene therapy for therapeutic angiogenesis in critically ischaemic lower limb - on the way to the clinic. AB - Currently, no effective pharmacological treatment is available for vascularisation defects in lower limbs. Many patients presenting with persistent pain and ischaemic ulcers are not suitable candidates for surgical or endovascular approaches. Further refinement of the available methods will undoubtedly lead to a more active approach towards treatment of peripheral arterial occlusive disease (PAOD). Recently, therapeutic angiogenesis, in the form of recombinant growth factor administration or gene therapy, has emerged as a novel tool to treat these patients. However, improved gene transfer methods and better understanding of blood vessel formation are required to bring therapeutic angiogenesis to clinical practice. Here we review the clinical problem (PAOD), mechanisms of blood vessel formation (angiogenesis, vasculogenesis and arteriogenesis), experimental evidence and clinical trials for therapeutic angiogenesis in critically ischaemic lower limbs. Also, angiogenic growth factors, including vascular endothelial growth factors (VEGFs) and fibroblast growth factors (FGFs), delivery methods, and vectors for gene transfer in skeletal muscle, are discussed. In addition to vascular growth, gene transfer of growth factors may enhance regeneration, survival, and innervation of ischaemic skeletal muscle. Nitric oxide (NO) appears to be a key mediator in vascular homeostasis and growth, and a reduction in its production by age, hypercholesterolemia or diabetes leads to the impairment of ischaemic disorders. PMID- 11473567 TI - Homocysteine levels in patients treated with lipid apheresis: effect of a vitamin therapy. AB - BACKGROUND: Patients treated with lipid apheresis already suffer from familial hypercholesterolemia and severe coronary heart disease: any additional risk factor is dangerous for these patients. Hyperhomocysteinemia has been recognized as an independent risk factor for atherosclerotic disease. We checked the frequency of hyperhomocysteinemia in lipid apheresis patients and measured the effect of a vitamin therapy. MATERIALS AND METHODS: Sixteen heterozygous patients (10 males, 6 females) were studied, who were being treated by three different apheresis procedures. Homocysteine was measured using an enzyme conversion immunoassay. Cystathionine and methylmalonic acid were assessed by gas chromatography/mass spectrometry. Serum levels of folic acid, vitamin B12, and vitamin B6 were also determined. The patients received a vitamin therapy (3 mg folate, 60 microg cyanocobalamine, 10 mg pyridoxine hydrochloride daily) for 12 weeks. RESULTS: In 9 out of 16 patients, plasma homocysteine levels were found to be elevated (> 12 micromol L(-1)). Cystathionine concentrations were also increased, especially in those patients with elevated homocysteine. Methylmalonic acid levels were not elevated. Serum folic acid, vitamin B6, and vitamin B12 concentrations were initially in the normal range and not correlated to plasma homocysteine. The vitamin therapy reduced the plasma homocysteine concentrations in all patients significantly by 33%. Among those patients with elevated homocysteine levels, the optimal range < 12 micromol L(-1) for homocysteine was rarely reached. CONCLUSIONS: In patients treated with lipid apheresis, a hyperhomocysteinemia can be frequently seen. The constellation of both elevated homocysteine and cystathionine levels points to the existence of tissue vitamin deficiencies, folate and vitamin B-6, which were improved by vitamin supplements. Because methylmalonic acid was mostly normal, a vitamin B-12 deficiency was not proven. PMID- 11473568 TI - Characterization of platelet-derived growth factor-induced p38 mitogen-activated protein kinase activation in vascular smooth muscle cells. AB - BACKGROUND: The mitogen-activated protein (MAP) kinase super-family plays a crucial role in cell growth and differentiation and even in programmed cell death in response to diverse extracellular stimuli. The platelet-derived growth factor (PDGF)-BB is well known to promote the proliferation of vascular smooth muscle cells (VSMC) via extracellular-regulated protein kinases (ERKs), leading to the development of cardiovascular diseases. However, it has not yet been clarified whether PDGFs that include other isoforms can activate the other parallel signal transduction pathways, c-jun NH2-terminal protein kinase (JNK) and p38 MAP kinase (p38), in VSMC. In this study, we investigated the effect of PDGFs on p38 activation in cultured rat VSMC. MATERIALS AND METHODS: After stimulation by PDGFs with SB-203580 or PD-98059, the cells were solubilized, and the expressions of MAP kinases, MAP kinase kinases (MKKs), phosphorylated DNA-binding proteins, and cyclooxigenases (COXs) were examined by immunoblot analysis. RESULTS: PDGFs activated p38 phosphorylation dose-dependently, and the phosphorylations were specifically inhibited by SB-203580 but not by PD-98059. PDGFs also activated the phosphorylation of MKK 3/MKK 6 but not that of either stress-activated protein kinase/ERK kinase or JNK. PDGFs affected the activation of a cyclic AMP response element binding protein, which was inhibited by SB-203580. However, the activating transcription factor-2 was not activated by PDGFs. Interestingly, the stimulation of PDGFs for 72 h enhanced the level of COX-2, and these levels were decreased by SB-203580. CONCLUSION: These results have clarified that PDGFs activate the p38 cascade via an MKK 3/6 pathway, independently of the ERK cascade, and subsequently regulate the level of COX-2 in rat VSMC, providing that PDGFs influence the inflammatory process in the vascular wall. PMID- 11473569 TI - Cardiovascular effects of beta 3-adrenoceptor stimulation in perinephritic hypertension. AB - BACKGROUND: A new beta 3-adrenoceptor (beta3-AR) has been shown to mediate peripheral vasodilation. This study was conducted to evaluate effects of the beta3-AR agonist, SR58611 in normal and hypertensive dogs. MATERIALS AND METHODS: In protocol 1, SR58611 was infused in normal dogs after placebo, after beta1/beta2 blockade with nadolol, after beta1/beta2/beta3 blockade with bupranolol and after combined autonomic blockade (CAB). In protocol 2, perinephritic hypertension was produced in dogs, which received SR58611 at 3 and 6 weeks of hypertension. Effects of SR58611 were evaluated at 7 weeks of hypertension after CAB. RESULTS: In normal dogs, SR58611 produced a dose dependent decrease in mean aortic pressure (AOP) (from 116 +/- 19 to 100 +/- 19 mmHg, - 14%; P < 0.05) that was accompanied by baroreflex activation (heart rate increased by 70%; P < 0.01). This hypotensive effect resulting from peripheral vasodilation persisted after nadolol or CAB while baroreflex activation was blunted or abolished. A biphasic response of cardiac output, characterized by a rise and a decline (P < 0.05) reflected a reduction in after- and pre-load. After CAB, SR58611 did not modify cardiac contractility. SR58611 stimulated lipolysis as reflected by a 4-fold increase in blood free fatty acids (FFA) (P < 0.0005). Under CAB, the rise of FFA was reduced (P < 0.01). In hypertensive dogs, SR58611 produced a dose-dependent decrease in mean AOP (from 168 +/- 32 to 125 +/- 35 mmHg; - 26%, P < 0.0001), that was greater than in normal dogs (P < 0.05). Reflex mediated tachycardia also occurred but at higher blood pressure values. Blood FFA rose similarly (P < 0.0001). Under CAB, heart rate remained unchanged but SR58611 still induced a decrease (P < 0.0001) in mean AOP concomitantly with a rise of (dP/dt)/DP40 (P < 0.005), an effect not observed in normal dogs. CONCLUSIONS: Beta3-AR stimulation exerts hypotensive effects, increases cardiac contractility and stimulates lipolysis in hypertensive dogs. PMID- 11473570 TI - Assessment of the left ventricular diastolic reserve in essential hypertension: the acute saline load test. AB - OBJECTIVE: The aim of this study was to evaluate the significance of the development of a restrictive response to an acute saline load, defined as an increase in the ratio of peak early to peak late diastolic transmitral flow velocity (E/A ratio) associated with a decrease in the deceleration time, in patients with mild to moderate untreated hypertension. BACKGROUND: Recognised abnormal patterns of transmitral diastolic flow include, from 'best' to 'worst': prolonged relaxation, pseudonormalisation, and restrictive physiology. The common denominator of these transitions is the constellation of an increase in the E/A ratio associated with a decrease in deceleration time. PATIENTS AND METHODS: Sixteen normal control subjects (6 males, 10 females, age 51.6 +/- 6.9 years) and 24 patients with mild to moderate untreated hypertension (12 males, 12 females, age 46.8 +/- 7.5 years) underwent supine blood pressure measurement with sphygmomanometry, biochemical studies, and transthoracic M-mode, 2D, and Doppler echocardiography before and after an acute saline load (7 mL kg(-1), maximum 500 mL, NaCl 0.9% within 15 min IV). RESULTS: The baseline E/A ratio was lower (0.90 +/- 0.14 vs. 1.04 +/- 0.18; P < 0.01) and the deceleration time was longer (158.8 +/- 19.4 vs. 135 +/- 8.9 ms; P < 0.01) in patients with hypertension compared with normotensive controls. However, no patient with hypertension exhibited a transmitral flow velocity pattern compatible with typical prolonged relaxation. A restrictive response to the acute saline load was observed in 12 (50%) of the hypertensive and none of the control subjects. Hypertensive patients with a restrictive response to the acute saline load had a lower baseline E velocity (54.8 +/- 8.7 cm s(-1) vs. 66 +/- 6.4 cm s(-1); P = 0.003), a lower baseline E/A ratio (0.83 +/- 0.13 vs. 0.97 +/- 0.12; P = 0.015), and a longer deceleration time (167.5 +/- 15.4 ms vs. 150 +/- 19.5; P = 0.03) than hypertensive patients without such a response. CONCLUSION: A restrictive response to an acute saline load is indicative of a limited diastolic reserve in patients with mild to moderate untreated hypertension. Further studies are required in order to evaluate the significance of such a response with regards to risk stratification and efficacy of medical treatment in this patient population. PMID- 11473571 TI - Comparison of angiotensin II receptor antagonists. AB - Nonpeptide orally active angiotensin II type 1 (AT1) receptor antagonists are the most specific means presently available to block the renin-angiotensin enzymatic cascade. Six of these drugs have already been licensed in Europe and in the United States for the treatment of high blood pressure, and additional candidates are in the pipeline. The World Health Organisation has also recently endorsed their use for this condition. Inasmuch as AT1 receptor antagonists have proven themselves the equals of angiotensin converting enzyme inhibitors with respect to antihypertensive efficacy, but demonstrated better safety profiles, this class of drugs may be considered to be a qualitative improvement in the treatment of essential hypertension. Interestingly, the six agents now on the market diverge considerably with respect to their pharmacokinetic and pharmacodynamic properties, although it is not certain whether such differences are clinically relevant. A considerable number of large, multicentre trials are in progress to ascertain the possible longer-term organoprotective effects of these substances on cardiovascular morbidity and mortality. Because of their noteworthy safety record to date, and simple once-a-day dosage regimen, AT1 receptor antagonists have the potential to improve compliance in patients with chronic hypertension. PMID- 11473572 TI - Gemfibrozil treatment potentiates oxidative resistance of high-density lipoprotein in hypertriglyceridemic patients. AB - BACKGROUND: Studies suggest that both oxidized low and high density lipoprotein (LDL and HDL) play a role in the pathogenesis of atherosclerosis. Gemfibrozil is widely used and is reported to increase cholesterol of LDL and HDL in hypertriglyceridemic patients. The aim of this study was to investigate the effect of gemfibrozil treatment on the oxidative status of lipoprotein particles in Fredrickson phenotype IV hypertriglyceridemic patients. METHODS: Twenty-two patients, aged 38-64 years, with fasting plasma triglyceride concentrations between 2.90 and 8.97 mmol L(-1), were recruited and were given gemfibrozil 300 mg three times daily for 12 weeks. Venous blood samples were collected before gemfibrozil treatment, after 4, 8, or 12 weeks of treatment, and 4 weeks after termination of treatment, and used to analyse the plasma lipid profile, isolate lipoproteins, and analyse the chemical composition and in vitro oxidation of lipoprotein particles. RESULTS: Gemfibrozil treatment resulted in a decrease in plasma total triglyceride levels and the triglyceride content of all lipoproteins. Plasma total cholesterol levels were decreased as a result of a decrease in very low density lipoprotein (VLDL) cholesterol levels. A slight increase in LDL cholesterol levels was observed, whereas the thiobarbituric acid reactive substances (TBARS) of LDL were decreased and the lag and peak time of LDL to oxidation were unchanged and maximal diene production was decreased. Plasma HDL cholesterol levels, the surface-to-core ratio of HDL particles, and the resistance of HDL to oxidation were increased. CONCLUSION: The decreased TBARS and diene production of LDL, increased HDL cholesterol levels, and increased resistance of HDL to oxidation may, in part, explain why gemfibrozil treatment was found to be generally beneficial in terms of protection against coronary heart disease. PMID- 11473573 TI - Fibroblast studies documenting a case of peroxisomal 2-methylacyl-CoA racemase deficiency: possible link between racemase deficiency and malabsorption and vitamin K deficiency. AB - BACKGROUND: 2-Methylacyl-CoA racemase interconverts the 2-methyl group of pristanoyl-CoA or the 25-methyl group of hydroxylated cholestanoyl-CoAs, allowing further peroxisomal desaturation of these compounds in man by the branched chain acyl-CoA oxidase, which recognise only the S-isomers. Hence, oxidation studies in fibroblasts, currently based on the use of racemic substrates such as [1-14C] pristanic acid, do not allow us to distinguish between a deficient racemase or an impaired oxidase. DESIGN: To evaluate the racemase activity directly, the 2R isomer of[1-14C] pristanic acid, as well as the 2R-isomer of 2-methyl-[1-14C] hexadecanoic, a synthetic pristanic acid substitute, were prepared and their degradation by cultured human skin fibroblasts was compared to that of the racemic substrates. RESULTS: In fibroblasts in a young girl, presenting with elevated urinary levels of trihydroxycholestanoic acid metabolites but normal plasma levels of very long chain fatty acids, a partial deficient degradation of racemic [1-14C] pristanic acid was observed. Incorporation of 2R-[1-14C] pristanic acid in glycerolipids of the patient's fibroblasts proceeded normally, but breakdown was impaired. Similar findings were seen with the 2R-isomer of 2 methyl-[1-14C] hexadecanoic. These data, combined with the fact that the branched chain acyl-CoA oxidase, catalyzing the first oxidation step of pristanic acid and bile acid intermediates in man, appeared normal, suggested a peroxisomal beta oxidation defect in the patient at the level of 2-methylacyl-CoA racemase. CONCLUSION: Carboxy-labelled 2R-methyl branched chain fatty acids might be useful tools to document cases of racemase deficiencies. Because a brother of the patient died with a diagnosis of vitamin K deficiency, an impaired racemase might be responsible for other cases of unexplicable malabsorption. PMID- 11473574 TI - Protein adducts in type I and type II fibre predominant muscles of the ethanol fed rat: preferential localisation in the sarcolemmal and subsarcolemmal region. AB - BACKGROUND: Chronic alcoholic myopathy is characterised by reduced muscle strength and structural changes including a decrease in the diameter of Type II (glycolytic, fast-twitch, anaerobic) fibres. In contrast, the Type I fibres (oxidative, slow-twitch, aerobic) are relatively protected. It is possible that adduct formation with reactive metabolites of ethanol may be a contributory process. MATERIALS AND METHODS: We analysed skeletal muscles from rats fed nutritional-complete liquid diets containing ethanol as 35% of total dietary energy; control rats were fed the same diet in which ethanol was replaced by isocaloric glucose. Reduced-acetaldehyde, unreduced-acetaldehyde, malondialdehyde, malondialdehyde-acetaldehyde and alpha-hydroxyethyl protein adducts in both soleus and plantaris were analysed by ELISA or immunohistochemistry with comparative studies in liver. RESULTS: After 6 weeks, the weights of the plantaris, but not the soleus, were decreased. ELISA analyses for protein adducts showed increased amounts of unreduced-acetaldehyde adducts in soleus (P < 0.025) and plantaris (P < 0.025). Reduced-acetaldehyde, malondialdehyde, malondialdehyde-acetaldehyde and alpha-hydroxyethyl protein adducts in both soleus and plantaris muscles from ethanol-fed rats were not significantly different from their pair-fed controls (P > 0.05). In contrast, liver from ethanol-fed rats showed significantly higher levels of unreduced acetaldehyde (P < 0.025), reduced-acetaldehyde (P < 0.01), malondialdehyde (P < 0.01), malondialdehyde-acetaldehyde (P < 0.025) and alpha-hydroxyethyl radical (P < 0.01) protein adducts compared to pair-fed controls. Immuno-histochemical analysis using an antiserum reacting with both reduced- and unreduced acetaldehyde adducts showed adducts were increased in soleus (P < 0.05) and plantaris (P < 0.025), confirming ELISA analysis. Adducts were located within the sarcolemmal (i.e. muscle membrane) and subsarcolemmal regions. CONCLUSION: This is the first report of adduct formation in myopathic skeletal muscle due to chronic alcohol ingestion and suggests a role for acetaldehyde in the aetiology of alcoholic myopathy. PMID- 11473575 TI - Relationship between bone mass density and tensile strength of the skin in women. AB - BACKGROUND: Skin and bone may be affected similarly during the climacteric and during long-term corticotherapy. Little is known about the correlation between the respective alterations in bone mass density (BMD) and tensile strength of the skin. MATERIALS AND METHODS: A total of 100 women aged 19-88 years, receiving, or not, hormone replenishment therapy or systemic corticosteroids, were enrolled in the study. Tensile strength of the inner forearm skin was measured using the suction method operated in both the steep and progressive-force application modes. BMD was measured by dual X-ray absorptiometry at the hip, femoral neck and lumbar spine. RESULTS: Being heterogeneous, the population of volunteers yielded a wide range of BMD and cutaneous tensile strength values. However, significant correlations were found between BMD and tensile skin parameters. In particular, a positive correlation was yielded between the biological elasticity of skin and the BMD of the hip and femoral neck. CONCLUSION: An overall correlation is shown between skin elasticity on a relatively sun-protected area and cortico-trabecular BMD. PMID- 11473576 TI - Induction and treatment of the antiphospholipid syndrome--lessons from animal models. AB - The diagnosis of antiphospholipid syndrome (APS) is based on a combination of clinical and serological criteria. This review summarizes the literature regarding animal models of APS. Animal models of this syndrome help to elucidate pathogenic roles of the characteristic antiphospholipid antibodies in pregnancy morbidity, vascular thrombosis, neurologic disease and atherosclerosis. Furthermore, these models also provide measures to evaluate novel therapies for APS. These include bone-marrow transplantation, intravenous immunoglobulin, anticoagulation, interleukin-3, in addition to other therapies. Experimental induction and treatment of APS largely contributes to our understanding of the disease and to better management of its clinical characteristics. PMID- 11473577 TI - AAA+ superfamily ATPases: common structure--diverse function. AB - The AAA+ superfamily of ATPases, which contain a homologous ATPase module, are found in all kingdoms of living organisms where they participate in diverse cellular processes including membrane fusion, proteolysis and DNA replication. Recent structural studies have revealed that they usually form ring-shaped oligomers, which are crucial for their ATPase activities and mechanisms of action. These ring-shaped oligomeric complexes are versatile in their mode of action, which collectively seem to involve some form of disruption of molecular or macromolecular structure; unfolding of proteins, disassembly of protein complexes, unwinding of DNA, or alteration of the state of DNA-protein complexes. Thus, the AAA+ proteins represent a novel type of molecular chaperone. Comparative analyses have also revealed significant similarities and differences in structure and molecular mechanism between AAA+ ATPases and other ring-shaped ATPases. PMID- 11473578 TI - BIP, a BRAM-interacting protein involved in TGF-beta signalling, regulates body length in Caenorhabditis elegans. AB - BACKGROUND: The TGF-beta superfamily has diverse biological activities and is involved in the early development of animals. We previously identified a novel family member, BMP receptor associated molecule (BRAM), which binds to the intracellular domain of BMP type IA receptor and is involved in the BMP signalling pathway. RESULTS: To identify novel molecules involved in TGF-beta signalling pathways, we performed yeast two-hybrid screening using BRAM as bait. From a Xenopus cDNA library, we cloned a cDNA encoding 693 amino acids and containing the motif for an oxysterol binding protein (OSBP), which we designated BRAM interacting protein (BIP). We then isolated a BIP homologue from the Caenorhabditis elegans that encodes 733 amino acids and also contains the OSBP like motif. Immunoprecipitation and Western blotting studies revealed that C. elegans BIP could interact with the C. elegans BRAM homologues BRA-1 and BRA-2. C. elegans BIP was expressed in pharyngeal muscle, hypodermis and several neuronal cells, an expression pattern overlaps with those of BRA-1 and BRA-2. Finally, we found that inhibition of BIP expression in C. elegans by double stranded RNA interference produces a Sma phenotype. CONCLUSIONS: BIP was isolated using the yeast two-hybrid systems. BIP may function in the TGF-beta pathway and regulate body length in C. elegans. PMID- 11473579 TI - Arabidopsis transcriptional regulation by light stress via hydrogen peroxide dependent and -independent pathways. AB - BACKGROUND: High (intense) light stress causes the formation of oxygen radicals in chloroplasts and has the potential to damage them. However, plants are able to respond to this stress and protect the chloroplasts by various means, including transcriptional regulation in the nucleus. Although the corresponding signalling pathway is largely unknown, the high light response in the expression of the Arabidopsis APX2 gene is reported to be mediated by hydrogen peroxide. RESULTS: We characterized light stress signalling by analysing expression profiles of another high light-inducible gene of Arabidopsis, ELIP2, with the aid of an ELIP2 promoter-luciferase gene fusion. The established ELIP2:LUC transgenic Arabidopsis showed activation by high light, but not by hydrogen peroxide. On the other hand, the native ELIP2 gene as well as the APX2 gene was activated by the hydrogen peroxide. The activation of ELIP2:LUC by intense light was not inhibited by K252a but by okadaic acid. CONCLUSION: The light stress signalling from the chloroplast to the nucleus is revealed to be mediated through at least two pathways: both hydrogen peroxide-dependent and -independent. The latter pathway is thought to be mediated by the protein phosphatase 2A/1 activity that is suppressed by okadaic acid. PMID- 11473580 TI - Ypel1: a novel nuclear protein that induces an epithelial-like morphology in fibroblasts. AB - BACKGROUND: Embryonic development depends on the regulation of cell morphology and behaviour to carry out morphogenesis. One example of this is the development of the face, which is constructed from independent blocks of tissue that must grow in a coordinated way and then fuse to form a continuous tissue. RESULTS: We have isolated a novel gene of unknown function from mouse and quail embryos in a search for genes involved in craniofacial development. Sequence analysis of this gene, known as Ypel1, demonstrates a striking level of amino acid conservation between vertebrates and invertebrates but no significant homology with any other characterized genes. Ypel1 is expressed in the ventral half of early embryos including the branchial arches from which the face derives. Ypel1 localizes to the nucleus, and transfection into fibroblasts induces an epithelial-like transition, which is accompanied by alterations to the cytoskeleton and cell adhesion machinery. In addition, human YPEL1 localizes to chromosome 22q11.2, a region associated with a number of syndromes involving malformation of the craniofacial complex. CONCLUSION: These data suggest a role in regulation of cellular morphology and behaviour that is important for development of the craniofacial complex. PMID- 11473581 TI - Cross talk of the interferon-alpha/beta signalling complex with gp130 for effective interleukin-6 signalling. AB - BACKGROUND: Signalling cross talk provides a molecular basis for modulating a given signalling pathway by another, and it is often critical for regulating cellular responses elicited by cytokines. Previously, we reported on the critical role of the IFN-alpha/beta signalling complex, generated by spontaneously produced IFN-alpha/beta, in efficient IFN-gamma signalling. RESULTS: In the present study, we have demonstrated that the IFN-alpha/beta signalling complex also contributes to efficient IL-6 signalling. In fact, IL-6-induced activation of the Stat1 and Stat3 transcription factors is markedly diminished in the absence of the IFN-alpha/beta signalling complex. The induction of several target genes for these factors is also diminished, both in vitro and in vivo. We provide evidence that the cytoplasmic tyrosine residues of IFNAR-1, which remains phosphorylated by a weak IFN-alpha/beta stimulation, provide docking sites for Stat1 and Stat3 to form homo- or heterodimers following IL-6 stimulation. Furthermore, a chemical cross-linking experiment revealed that IFNAR-1 and gp130, a common signal transducer for the IL-6 family of cytokines, exist in close proximity. CONCLUSIONS: The constitutive weak IFN-alpha/beta signal provides a foundation for strong cellular responses to IL-6, IFN-gamma, and possibly other cytokines. Our results also suggest the assembly of cytokine receptor subunits, which may represent a 'receptosome'-like structure, allowing the unique signalling cross talks to occur. PMID- 11473582 TI - Terminal deoxynucleotidyltransferase directly interacts with a novel nuclear protein that is homologous to p65. AB - BACKGROUND: Terminal deoxynucleotidyltransferase (TdT) is a DNA polymerase that enhances Ig and TcR gene diversity in the N region in B- and T-cells. TdT is found as a member of a large protein complex in the lysate of the thymocytes. To elucidate the molecular mechanism of the synthesis of the N region, we first attempted to isolate the genes with products that are interacting directly with TdT. RESULTS: Using a yeast two-hybrid system, we isolated a cDNA clone encoding a novel nuclear protein that interacts with TdT. This protein was designated as TdT interacting factor 1 (TdIF1). TdIF1 has a high degree of homology to the transcription factor p65, which belongs to the nuclear receptor superfamily. TdIF1 contains HMG-I and HMG-Y DNA binding domains (AT-hooks) and can bind to single- and double-stranded DNA. TdT and TdIF1 were co-eluted at position 232 kDa by gel filtration of MOLT4 lysate. TdIF1 can enhance TdT activity fourfold in vitro assay system using oligo(dT)16 as primers. CONCLUSIONS: TdIF1 binds directly to TdT, both in vitro and in vivo. TdIF1 and TdT exist as the members of a 232 kDa protein complex. TdIF1 can enhance TdT activity maximum fourfold in vitro assay system, suggesting that it positively regulates the synthesis of the N region during V(D)J recombination in the Ig and TcR genes. PMID- 11473583 TI - Developments in microbiological risk assessment for drinking water. PMID- 11473584 TI - Coinfection with Campylobacter species: an epidemiological problem? AB - AIMS: To determine the frequency of coinfection with multiple strains in sporadic cases of human Campylobacter infection. METHOD AND RESULTS: During 1999 10 single colonies of Campylobacter were cultured from each of 53 positive faecal samples. Five isolates were taken from nonselective agar after passive filtration of faecal suspensions and five isolates were taken from selective agar plates. All isolates were sero- and phage typed and their antibiotic resistance determined. Pulsed-field gel electrophoresis and flagellin gene typing were performed on selected isolates. One patient was infected with Camp. coli, the remainder with strains of Camp. jejuni. The majority of patients was infected with a single strain of Campylobacter, but from each of four samples, 7.5%, two strains of Camp. jejuni, confirmed by molecular typing, were identified. CONCLUSION: Coinfection occurs in sporadic cases of campylobacteriosis. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has implications in outbreak investigation when distinct strains have been isolated from epidemiologically related patients and/or the suspected source or vehicle. PMID- 11473585 TI - Disinfection of wooden structures contaminated with Paenibacillus larvae subsp. larvae spores. AB - AIMS: The aim of the study is to examine the disinfection of wood contaminated with Paenibacillus larvae subsp. larvae spores, in order to find a practical decontamination method for hive materials. METHODS AND RESULTS: The number of viable spores recovered after the treatment, on the surface by swabbing, and in the deeper parts of the wood by scraping, was used to test the efficiency of the disinfection. Our results indicate that chemical disinfection is only complete when high concentrations (> 50%) of the disinfectant are used. Heat treatment in general was found to be very effective. The scorching of wood was not satisfactory as it only killed spores at the surface. CONCLUSION: Complete disinfection is only possible with some heat treatments or by using high concentrations of chemical disinfectants. SIGNIFICANCE AND IMPACT OF THE STUDY: This study puts forward some methods that can provide complete decontamination, which is necessary for an effective control of American foulbrood disease. PMID- 11473586 TI - The ability of Fla-typing schemes to discriminate between strains of Campylobacter jejuni. AB - AIMS: The aim of this investigation was to compare the usefulness of two previously published flagellin PCR-RFLP typing (Fla-typing) techniques for the subtyping of Campylobacter jejuni strains, in terms of ease of use and discriminatory power. METHODS AND RESULTS: Six groups of isolates, which were epidemiologically unrelated but with similar Fla-types, and five groups of epidemiologically related poultry isolates, with similar PFGE profiles, were used in the comparison. The Fla-typing methods used varied in the number and length of fla-genes amplified and the restriction enzymes used. In addition, the use of separately amplified PCR fragments of both the flaA and flaB genes to generate RFLP profiles was investigated. CONCLUSION: The results clearly demonstrated that both previously published methods exhibit some advantages over the other. However, optimal discrimination was obtained by the use of separately amplified PCR fragments of both fla-genes. SIGNIFICANCE AND IMPACT OF THE STUDY: The subtyping of Camp. jejuni isolates is considered essential for epidemiological purposes. Genotyping methods are now more frequently used but have yet to be standardized. Fla-typing is a rapid and easy to use method with acceptable discriminatory power. However, the discriminatory power of the currently published Fla-typing techniques may be further improved by incorporating RFLP profiles of both fla-genes. PMID- 11473587 TI - Comparison of pattern recognition techniques for the identification of lactic acid bacteria. AB - AIMS: The goal of this study was to evaluate three pattern recognition methods for use in the identification of lactic acid bacteria. METHODS AND RESULTS: Lactic acid bacteria (21 unknown isolates and 30 well-characterized strains), including the Lactobacillus, Lactococcus, Streptococcus, Pediococcus and Oenococcus genera, were tested for 49 phenotypic responses (acid production on carbon sources). The results were scored in several ways. Three procedures, k nearest neighbour analysis (KNN), k-means clustering and fuzzy c-means clustering (FCM), were applied to the data. CONCLUSION: k-Nearest neighbour analysis performed better with five-point-scaled than with binary data, indicating that intermediate values are helpful to classification. k-Means clustering performed slightly better than KNN and was best with fuzzified data. The best overall results were obtained with FCM. Genus level classification was best with FCM using an exponent of 1.25. SIGNIFICANCE AND IMPACT OF THE STUDY: The three pattern recognition methods offer some advantages over other approaches to organism classification. PMID- 11473588 TI - Modelling bacterial spoilage in cold-filled ready to drink beverages by Acinetobacter calcoaceticus and Gluconobacter oxydans. AB - AIMS: Mathematical models were created which predict the growth of spoilage bacteria in response to various preservation systems. METHODS AND RESULTS: A Box Behnken design included five variables: pH (2.8, 3.3, 3.8), titratable acidity (0.20%, 0.40%, 0.60%), sugar (8.0, 12.0, 16.0 * Brix), sodium benzoate concentration (100, 225, 350 ppm), and potassium sorbate concentration (100, 225, 350 ppm). Duplicate samples were inoculated with a bacterial cocktail (100 microl 50 ml(-1)) consisting of equal proportions of Acinetobacter calcoaceticus and Gluconobacter oxydans (5 x 10(5) cfu ml(-1) each). Bacteria from the inoculated samples were enumerated on malt extract agar at zero, one, two, four, six, and eight weeks. CONCLUSION: The pH, titratable acidity, sugar content, sodium benzoate, and potassium sorbate levels were all significant factors in predicting the growth of spoilage bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This beverage spoilage model can be used to predict microbial stability in new beverage product development and potentially reduce the cost and time involved in microbial challenge testing. PMID- 11473589 TI - Assessment of resistance towards biocides following the attachment of micro organisms to, and growth on, surfaces. AB - AIMS: To develop a rapid method for the assessment of biocidal activity directed towards intact biofilms. METHODS AND RESULTS: Escherichia coli and Staphylococcus epidermidis were cultured for up to 48 h within 96-well microtitre plates. The planktonic phase was removed and the wells rinsed. Residual biofilms were exposed to various concentrations of chloroxylenol, peracetic acid, polyhexamethylene biguanide (PHMB), cetrimide or phenoxyethanol for 1 h. At 15-min intervals, biocide was removed, and the wells washed in neutraliser and filled with volumes of fresh medium. Re-growth of the cultures was monitored during incubation at 35 degrees C in the plate reader. Times taken for the treated wells to re-grow to fixed endpoints were determined and related to numbers of surviving cells. Time- survival curves were constructed and the survival of the attached bacteria, following exposure to the agents for 30 min, interpolated for each biocide concentration. Log--log plots of these survival data and biocide concentration were constructed, and linear regression analysis performed in order to (i) calculate concentration exponents and (ii) compare the effectiveness of the biocides between variously aged biofilm and planktonic cells. From such analyses iso-effective concentrations of biocide (95% kill in 30 min) were calculated and expressed as planktonic : biofilm indices (PBI). CONCLUSION: PBI varied between 1.02 and 0.02, were relatively unaffected by age of the biofilms but differed significantly between organism and biocide. Notably those compounds with the higher activity against planktonic bacteria (PHMB and peracetic acid) were most prone to a biofilm effect but remained the most effective of the agents selected. SIGNIFICANCE AND IMPACT OF THE STUDY: The endpoint method proved robust, enabled the bactericidal effects of the biocides to be assessed against in-situ biofilms, and was suitable for routine screening applications. PMID- 11473590 TI - Campylobacter contamination in French chicken production from farm to consumers. Use of a PCR assay for detection and identification of Campylobacter jejuni and Camp. coli. AB - AIMS: Campylobacter contamination in French chicken production from the farm to the consumer was determined using a PCR assay for bacteria detection and identification. METHODS AND RESULTS: Samples were bird droppings from poultry houses, neck skins, livers, hearts, gizzards, wings, legs and escalopes from slaughterhouses and gizzards, legs, drumstick, breast and escalopes from a supermarket. Bacterial DNA extraction was performed after an enrichment step in a broth and was followed by PCR. An internal control (IC) was used for both DNA extraction and PCR. Campylobacter were detected in 79.2% of poultry houses. Of the 303 samples, 201 were Campylobacter-positive (i.e. 66.3%) including 43.2% faecal samples, 5.6% slaughterhouse samples and 17.5% supermarket samples. There was no significant difference between the molecular method and the conventional culture technique for Campylobacter detection whatever the samples. The sensitivity was 5 UFC g(-1) of samples and 1.5 x 10(3) UFC ml(-1) of enrichment broth. The use of IC revealed PCR inhibition in 13 samples and problems in the DNA extraction in five samples. CONCLUSION: Significant Campylobacter contamination affects all stages of French chicken production. SIGNIFICANCE AND IMPACT OF THE STUDY: The understanding of Campylobacter contamination at different levels of chicken production and the determination of the best place(s) for intervention are important for significantly decreasing Campylobacteriosis. Our technique is rapid and can be used on different chicken samples for Campylobacter detection and identification. PMID- 11473591 TI - Identification of plant-associated enterococci. AB - AIMS: Enterococcus isolates from forage grass were subjected to taxonomical investigations and tested for antibiotic resistance. METHODS AND RESULTS: The identification procedure included phenotypic characterizations, restriction analyses of polymerase chain reaction-amplified 16S rDNA, whole-cell protein profile analyses and 16S rDNA sequence analyses. Agar diffusion tests were performed to detect antibiotic resistance. CONCLUSION: The isolates were identified as belonging to the species Enterococcus faecium, Ent. mundtii, Ent. casseliflavus, Ent. faecalis and Ent. sulfureus. However, the majority of isolates differed distinctly in their restriction patterns from those of known species. They formed a group of a homogeneous 16S rDNA genotype (VI). The 16S rDNA sequence of a representative isolate revealed the closest relationship to the species Ent. faecalis (similarity of 97.4%). All isolates were sensitive to vancomycin, but almost all were resistant to gentamycin and streptomycin. SIGNIFICANCE AND IMPACT OF THE STUDY: The taxonomical investigations suggest that the isolates of the 16S rDNA genotype VI represent a new plant-associated Enterococcus species. PMID- 11473592 TI - A phylogenetic analysis of Bacillus thuringiensis serovars by RFLP-based ribotyping. AB - AIMS: To determine the 23S and 5S rRNA gene fingerprints in order to reveal phylogenetic relationships among Bacillus thuringiensis strains. METHODS AND RESULTS: Eighty-six B. thuringiensis strains which include 80 serovar type strains, five intraserovar strains and a non-serotypeable strain, wuhanensis, were tested. Total DNA was digested with EcoRI and HindIII. The 23S and 5S rRNA gene restriction fragment length polymorphisms showed 82 distinctive ribopatterns. The dendrogram generated by numerical analysis showed 10 phylogenetic groups and six ungrouped serovars at the 95.5% DNA relatedness rate. A second dendrogram was constructed using a combination of the data from this study and from a previous study on 16S rRNA gene fingerprinting. It revealed eight distinct phylogenetic groups and three ungrouped serovars at the 94% DNA relatedness rate. CONCLUSION: This method permitted the classification and positioning of a wide variety of B. thuringiensis strains on a phylogenetic tree. Bacillus thuringiensis strains appear to be relatively homogeneous and to share a high degree of DNA relatedness. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes a further step to the definition of valid taxonomic sublevels for the B. thuringiensis species. PMID- 11473593 TI - Studies on bioemulsifier production by Acinetobacter strains isolated from healthy human skin. AB - AIMS: In recent years, interest has been growing in the search for novel bioemulsifiers. Many bacterial genera including Acinetobacter have been reported to produce bioemulsifiers. The present study aims to screen Acinetobacter isolates from healthy human skin for bioemulsifier production. METHODS AND RESULTS: Acinetobacter junii SC14 produced maximum bioemulsifier in the presence of almond oil during stationary growth phase at 37 degrees C and pH 7.2. Partially purified, nondialysable bioemulsifier from SC14 was a proteoglycan. The protein and polysaccharide fractions resulted in 95.2% reconstitution of the emulsification activity. The role of esterase in the release of cell-bound emulsifier and the contribution of capsular polysaccharide to the emulsification activity were observed. CONCLUSION: Acinetobacter strains from human skin exhibited better emulsification activity than that by burn wound or soil isolates, owing to the inherent differences in chemical microenvironment of their habitats. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigation of skin commensals, especially acinetobacters, would lead to the discovery of novel bioemulsifiers with interesting properties. Attempts of screening and strain improvement directed towards skin commensals will open up new avenues for strains producing bioemulsifier on a commercial scale. PMID- 11473594 TI - Synthesis of intracellular storage polymers by Amaricoccus kaplicensis, a tetrad forming bacterium present in activated sludge. AB - AIMS: The study investigated the physiology of Amaricoccus kaplicensis to determine whether it could outcompete polyphosphate accumulating bacteria in activated sludge systems removing phosphorus, by preferentially assimilating substrates in the anaerobic stages of these processes. METHODS AND RESULTS: The storage processes were investigated under anaerobic, anoxic and aerobic conditions in both batch and periodically fed cultures in an aerobic sequencing batch reactor (SBR). Amaricoccus kaplicensis showed a high capacity for storing aerobically large amounts of acetate as poly beta-hydroxybutyrate (PHB) at high rates. However, no acetate assimilation under anaerobic conditions and very slow assimilation under anoxic conditions could be detected. CONCLUSION: Amaricoccus kaplicensis in pure culture does not behave as polyphosphate accumulating bacteria competitor; therefore it is difficult to understand why anaerobic/aerobic systems often contain such large numbers of Amaricoccus cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Amaricoccus kaplicensis is probably not responsible for the failure of activated sludge systems removing phosphorus, and other organisms capable of anaerobic substrate assimilation should be sought. PMID- 11473595 TI - The influence of dissolved CO(2) concentration on the death kinetics of Saccharomyces cerevisiae. AB - AIMS: The effects of temperature and concentration of dissolved CO(2) on the inactivation of Saccharomyces cerevisiae were investigated using a plug-flow system. METHODS AND RESULTS: Several combinations of pressure (4, 6, 8, 10 mega Pa (MPa)) and temperature (30, 34, 36, 38 degrees C) were used. The D-values obtained were 0.14 min at 8 MPa and 38 degrees C, and 0.15 min at 10 MPa and 36 degrees C. The log D-values were related linearly to the treatment temperature and to the dissolved CO(2) concentration. The thermal resistance constant (zCO(2)(T)) was 9.5 degrees C in the media, including significant levels of CO(2), and the CO(2) resistance constant was z(temp.)(gamma)=7.2 gamma. CONCLUSION: This work has shown that inactivation followed first-order death kinetics, and the effects of temperature and CO(2) concentration were consistent through the critical temperature and pressure of CO(2). Therefore, it is feasible to estimate D-values at any temperature and any CO(2) concentration. SIGNIFICANCE AND IMPACT OF THE STUDY: Non-thermal inactivation of micro-organisms in acidic beverages could be realized by the present technique. PMID- 11473596 TI - Amino acid catabolism in cheese-related bacteria: selection and study of the effects of pH, temperature and NaCl by quadratic response surface methodology. AB - AIMS: To screen the cystathionine lyase and L-methionine aminotransferase activities of cheese-related bacteria (lactococci, non-starter lactobacilli and smear bacteria) and to determine the individual and interactive effects of temperature, pH and NaCl concentration on selected enzyme activities. METHODS AND RESULTS: A subcellular fractionation protocol and specific enzyme assays were used, and a quadratic response surface methodology was applied. The majority of the strains, 21 of 33, had detectable cystathionine lyase activity which differed in the specificity. Aminotransferase activity on L-methionine was observed in only three strains. The cystathionine lyase activities of Lactobacillus reuteri DSM20016, Lactococcus lactis subsp. cremoris MG1363, Brevibacterium linens 10 and Corynebacterium ammoniagenes 8 and the L-methionine aminotransferase activity of Lact. reuteri DSM20016 had temperature and pH optima of 30-45 degrees C, and 7.5 8.0, respectively. As shown by the quadratic response surface methodology these enzymes retained activities in the range of temperature, pH and NaCl concentration which characterized the cheeses from which the bacteria originated. CONCLUSION: The enzyme activities may have a role in flavour development during cheese ripening. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of this work contribute to the knowledge about the amino acid catabolic enzymes in order to improve cheese ripening. PMID- 11473597 TI - Comparison of selective media for the detection of Vibrio vulnificus in environmental samples. AB - AIMS: To compare two selective agars, cellobiose-colistin (CC) agar and a modification of the Vibrio vulnificus medium (VVMc agar), for the isolation of Vibrio vulnificus from environmental samples. METHODS AND RESULTS: The efficiencies of recovery of V. vulnificus collection strains on CC, VVM, VVMc and on thiosulphate-citrate-bile salts-sucrose (TCBS) agar were compared and similar efficiencies were obtained. A slightly higher recovery was observed on VVMc agar. The detection of V. vulnificus in environmental samples (eels and water) was performed by combining culture-based methods (CC and VVMc agars) with DNA-based methods using species-specific probes based on the cytolysin-haemolysin and the 16S rDNA genes. A lower accompanying microbiota was found on CC agar than on VVMc agar. CONCLUSION: The comparison between CC and VVMc agars confirms that both are useful for the detection of V. vulnificus in environmental samples. However, the use of any of these media should be combined with a species-specific probe. SIGNIFICANCE AND IMPACT OF THE STUDY: The combined use of a selective medium and a specific probe provides a feasible method for the detection of V. vulnificus for epidemiological and ecological studies. PMID- 11473598 TI - Partial characterization of a bacteriocin produced by Lactobacillus delbrueckii subsp. lactis UO004, an intestinal isolate with probiotic potential. AB - AIMS: The partial characterization of a bacteriocin produced by a human Lactobacillus delbrueckii isolate with probiotic potential. METHODS AND RESULTS: A bacterocin, UO004, was partially purified by cation exchange followed by a hydrophobic interaction column, biochemically characterized and the N-terminal region sequenced. Bacteriocin UO004 was found to be a hydrophobic, heat-stable polypeptide with an apparent molecular mass of 6 kDa. It was also stable and active over a wide pH range. CONCLUSION: The active compound was proteinaceous, heat-stable, and had a bactericidal (and bacteriolytic) mode of action on a limited number of micro-organisms. Such a narrow spectrum of activity is typical for bacteriocins produced by intestinal Lactobacillus. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteriocin UO004 from a probiotic strain is a new compound that does not share any homology with any other known lactic acid bacteria bacteriocin. Furthermore, Lact. delbrueckii is regarded as a suitable starter for the production of fermented milks. PMID- 11473599 TI - Nucleotide sequence and characterization of the cell envelope proteinase plasmid in Lactococcus lactis subsp. cremoris HP. AB - AIMS: The major cell envelope proteinase (lactocepin; EC 3.4.21.96) produced by Lactococcus lactis cheese starter bacteria is required for starter growth and acid production in milk. The aim of this study was to characterize a lactocepin plasmid from a L. lactis subsp. cremoris cheese starter strain. METHODS AND RESULTS: A restriction map of the lactocepin plasmid pHP003 from strain HP was constructed, fragments were cloned in Escherichia coli vectors, and the complete DNA sequence (13,433 bp) was determined. Among 120 industrial L. lactis starter strains screened, five contained the same specificity-type lactocepin as pHP003. The lactocepin gene in these strains was invariably linked with a partially deleted abiB gene. CONCLUSION: The lactocepin specificity type of strain HP, conferred by a known configuration of key residues, is relatively uncommon. The gene is invariably linked with a partially deleted abiB gene on each lactocepin plasmid. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first complete sequence reported for a lactocepin plasmid, and provides the basis for better understanding, or manipulation, of lactocepin production. PMID- 11473600 TI - Methicillin resistance study in clinical isolates of coagulase-negative staphylococci and determination of their susceptibility to alternative antimicrobial agents. AB - AIMS: To achieve reliable detection of methicillin resistance in clinical isolates of coagulase-negative staphylococci. METHODS AND RESULTS: Strains (105) were evaluated by normatized antimicrobial susceptibility methods, and for the presence of the methicillin resistance-determining mecA gene, using the polymerase chain reaction. Correlation between phenotypic and genotypic methods was obtained in 87.6% of the samples. Six strains, classified as methicillin susceptible by phenotypic assays, revealed the presence of the mecA gene, indicating that methicillin resistance expression was probably repressed. Another seven isolates failed to show mecA amplification after displaying methicillin resistance in phenotypic evaluations. The susceptibility of the methicillin resistant isolates to other antimicrobial agents was variable. CONCLUSION: Genotypic determination of the mecA gene proved to be the most reliable method for detection of methicillin resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: Correct assessment of methicillin resistance, such as that attained through genotyping, is essential for defining therapeutic strategies, particularly when treating severely compromised patients. PMID- 11473601 TI - Advances in disinfection testing and modelling. AB - AIMS: To develop a set of kinetic equations which more ably describe the disinfection process. METHODS AND RESULTS: A group of functions, the fat equations, based on the model used for the quantification of microbial inhibition, was produced. These functions introduce a limit to the numbers of micro-organisms capable of being disinfected. These new expressions were shown to be more general forms of currently-used (e.g. log-linear) disinfection models, and accommodate the lags and/or tails of non-linear log-survivor--time plots. An advance in the experimental procedures used to obtain disinfection data, using an optical density technique, was developed concomitantly. CONCLUSION: The methods of analyses (experimental and modelling) allow the researcher to examine, more ably, five-minute disinfection (or specific time disinfection tests) as well as the more important disinfection rate analyses. SIGNIFICANCE AND IMPACT OF THE STUDY: The fat equations are an improvement over commonly-used rate models of disinfection, which are shown to be special cases of these equations. This raises the question as to whether our current understanding of the kinetic basis of disinfection requires revision. PMID- 11473602 TI - Analysis of the role of bacterial endospore cortex structure in resistance properties and demonstration of its conservation amongst species. AB - AIMS: The aim of this work was to compare the chemical structure of the spore cortex of a range of species, and to determine any correlation between cortex structure and spore resistance properties. METHODS AND RESULTS: The fine chemical structure of the cortex of Bacillus subtilis, Bacillus megaterium, Bacillus cereus and Clostridium botulinum was examined by muropeptide analysis using reverse phase HPLC. There is a conserved basic structure between peptidoglycan of these species, with the only difference being the level of de-N-acetylation of an amino sugar. In order to determine if an alteration in cortex structure correlates with heat resistance properties, the peptidoglycan structure and properties of B. subtilis spores prepared under different conditions were compared. Peptidoglycan from spores prepared in Nutrient Broth (NB) showed reduction in single L-alanine substituted muramic acid to only 13.9% compared with 20.6% in CCY-grown spores. NB-prepared spores are also unstable, with 161 fold less heat resistance (60 min, 85 degrees C) and 43 times less Mn(2+) content than CCY-grown spores. Addition of MnCl(2) to NB led to a peptidoglycan profile similar to CCY-grown spores, sevenfold more heat resistance (60 min, 85 degrees C) and an 86-fold increase in Mn(2+) content. Addition of CCY salts to NB led all parameters to be comparable with CCY-grown spore levels. CONCLUSION: It has been shown that peptidoglycan structure is conserved in four spore-forming bacteria. Also, spore heat resistance is multifactorial and cannot be accounted for by any single parameter. SIGNIFICANCE AND IMPACT OF THE STUDY: Endospores made by diverse species most likely have common mechanisms of heat resistance. However, the molecular basis for their resistance remains elusive. PMID- 11473603 TI - The optimization of isolation media used in immunomagnetic separation methods for the detection of Escherichia coli O157 in foods. AB - AIMS: To compare media used in immunomagnetic separation (IMS) techniques for the isolation of Escherichia coli O157 from food. METHODS AND RESULTS: Foods, both naturally contaminated and spiked, with low numbers (< 1 g(-1)) of stressed E. coli O157 were enriched in media based on buffered peptone water (BPW), tryptone soya and EC broths incubated at 30, 37, 40 and 42 degrees C. Following immunomagnetic separation, beads were plated on a range of selective agars. CONCLUSION: BPW supplemented with vancomycin (8 mg l(-1)) incubated at 42 degrees C, followed by IMS and subsequent plating of immunobeads onto cefixime tellurite sorbitol MacConkey agar plus either Rainbow or CHROMagar agars, proved optimum for the recovery of spiked, stressed E. coli O157 in minced beef, cheese, apple juice and pepperoni. The same protocol was optimum for recovery from naturally contaminated minced beef and cheese. SIGNIFICANCE AND IMPACT OF THE STUDY: The optimum protocol would increase isolation rates of E. coli O157 from foods. PMID- 11473617 TI - Transcriptional regulation by changes in tonicity. AB - Most organisms respond to a hypertonic environment by accumulating small organic solutes. In contrast to high concentrations of electrolytes, the small organic solutes do not perturb the activity of enzymes and other macromolecules within the cell. When the renal medulla becomes hypertonic during antidiuresis, multiple signaling pathways are activated. Here, we review the role of tonicity responsive enhancers (TonE) binding protein (TonEBP), a transcription factor activated in hypertonic cells. The activation of TonEBP by hypertonicity results from its translocation to the nucleus as well as an increase in TonEBP mRNA and protein. TonEBP may have a role beyond the response to tonicity since it is highly expressed in activated lymphocytes and in developing tissues. PMID- 11473618 TI - Transcriptional regulation of the NPT2 gene by dietary phosphate. AB - Dietary phosphate (Pi) is an important regulator for renal Pi reabsorption. The type II sodium-dependent phosphate (Na/Pi) cotransporters (NPT2) are located at the apical membranes of renal proximal tubular cells and major functional transporters associated with renal Pi reabsorption. The yeast one-hybrid system was used to clone a transcription factor that binds to a specific sequence (Pi response element) in the promoter of the NPT2 gene. Two cDNA clones that encoded protein of the mouse transcription factor mu E3 (TFE3) were isolated. TFE3 may participate in the transcriptional regulation of the NPT2 gene by dietary Pi. PMID- 11473619 TI - Isolation of a novel zinc finger repressor that regulates the kidney-specific CLC K1 promoter. AB - CLC-K1 and CLC-K2, two kidney-specific CLC chloride channels, are transcriptionally regulated on a tissue-specific basis. We have shown that a GA element near their transcriptional start sites is important for basal and cell specific activities of the CLC-K1 and CLC-K2 gene promoters. To identify the GA binding proteins, a kidney cDNA library was screened by a yeast one-hybrid system. A novel member of the Cys2-His2 zinc finger gene designated as KKLF (kidney-enriched Kruppel-like factor) and the myc-associated zinc finger protein (MAZ) were cloned. KKLF was found to be abundantly expressed in the liver, kidney, heart, and skeletal muscle. In the kidney, KKLF protein was localized in interstitial cells, mesangial cells, and nephron segments where CLC-K1 and CLC-K2 were not expressed. Gel mobility shift assay revealed that recombinant KKLF and MAZ proteins exhibited sequence-specific binding to the CLC-K1 GA element and that the consensus sequence for the KKLF binding site was GGGGNGGNG. In transient transfection, MAZ had a strong activating effect on the CLC-K1-luciferase reporter gene transcription. On the other hand, KKLF coexpression with MAZ appeared to block the activating effect of MAZ. These results suggest that a novel set of zinc finger proteins may help regulate the strict tissue and nephron segment-specific expression of CLC-K1 and CLC-K2 channel genes through their GA cis element. PMID- 11473620 TI - Oligomerization of water and solute channels of the major intrinsic protein (MIP) family. AB - Water and small solute fluxes through cell membranes are ensured in many tissues by selective pores that belong to the major intrinsic protein family (MIP). This family includes the water channels or aquaporins (AQP) and the neutral solute facilitators such as the glycerol facilitator (GlpF). We have compared the characteristics of representatives of each subfamily. Following solubilization in the nondenaturing detergents n-octyl-glucoside (OG) and Triton X-100 (T-X100), AQPs remain in their native homotetrameric state, while GlpF always behaves as a monomer. Solute facilitators are fully solubilized by the detergent N-lauroyl sarcosine (NLS), while AQPs are not. Analyses of mutants and chimeras demonstrate a close correlation between the water transport function and the resistance to NLS solubilization. Thus, AQPs and solute facilitators exhibit different behaviors in mild detergents; this could reflect differences in quaternary organization within the membranes. We propose that the oligomerization state or the strength of self-association is part of the mechanisms used by MIP proteins to ensure solute selectivity. PMID- 11473621 TI - Ion pump sorting in polarized renal epithelial cells. AB - The plasma membranes of renal epithelial cells are divided into distinct apical and basolateral domains, which contain different inventories of ion transport proteins. Without this polarity vectorial ion and fluid transport would not be possible. Little is known of the signals and mechanisms that renal epithelial cells use to establish and maintain polarized distributions of their ion transport proteins. Analysis of ion pump sorting reveals that multiple complex signals participate in determining and regulating these proteins' subcellular localizations. PMID- 11473622 TI - Molecular determinants for apical expression and regulatory membrane retrieval of the type IIa Na/Pi cotransporter. AB - Renal inorganic phosphate (Pi) reabsorption is a key process in Pi homeostasis. Type IIa Na/Pi cotransporters, located at the apical membrane of renal proximal tubular cells, guarantee the vectorial transport of Pi. Renal Pi reabsorption can be modulated by controlling the number of cotransporters expressed at the apical membrane. Indeed, factors that increase Pi reabsorption induce the expression of type IIa cotransporters at the apical membrane, whereas factors that decrease Pi reabsorption lead to their retrieval. Therefore, proper sorting of this type of cotransporters is an essential step in Pi homeostasis. The relevance of polarization has been highlighted by the finding that improper sorting of transporters can cause disease. Here we describe the identification of signals involved in apical expression of newly synthesized type IIa cotransporters and in their hormonal-induced endocytosis. PMID- 11473623 TI - Renal potassium channels: function, regulation, and structure. AB - Many transport functions in renal tubules depend on potassium (K) channels. Not only does K secretion and the maintenance of external K balance depend on K channel activity in principal tubule cells, but K channels also regulate cell volume; they are an integral party of cell function in all tubule cells because of their key role in the generation of the cell-negative electrical potential that affects the transmembrane movement of many charged solutes. Moreover, the recycling of K across the apical membrane of the thick ascending limb (TAL) plays an important role in the control of NaCl reabsorption in this tubule segment. Significant progress in our understanding of the structure and function of renal K channels has become possible by combining several strategies. These include transport studies in single tubules, application of the patch-clamp technique for exploring the properties of single K channels in native tubules and the cloning, and expression of diverse K channels of renal origin. Insights from these investigations promise to provide a deeper understanding of the mechanism by which K channels participate in many diverse tubule functions. PMID- 11473624 TI - Role and identification of protein kinase A anchoring proteins in vasopressin mediated aquaporin-2 translocation. AB - The antidiuretic hormone arginine vasopressin (AVP) regulates water reabsorption in renal principal cells by inducing a cAMP/protein kinase A-dependent translocation of water channels [aquaporin-2 (AQP2)] from intracellular vesicles into the apical cell membranes. Using primary cultured rat inner medullary collecting duct (IMCD) cells, it has been shown that AQP2 translocation in response to AVP stimulation occurs only if protein kinase A (PKA) is anchored to PKA anchoring proteins (AKAPs), which are present in various subcellular compartments. The identity of the AKAPs involved has not yet been elucidated. One potential candidate is a new splice variant of AKAP18, namely AKAP18 delta. PMID- 11473625 TI - Acute regulation of NHE3 by protein kinase A requires a multiprotein signal complex. AB - Biochemical and cellular experiments in fibroblasts have established the requirement for a member of the PDZ motif Na(+)/H(+) exchanger regulatory factor family of proteins (NHERF and NHERF2) in cAMP-mediated phosphorylation and inhibition of NHE3 activity. NHERF interacts with the actin cytoskeleton through the scaffolding protein ezrin to target a multiprotein signal complex to the plasma membrane. Recent experiments have focused on elements of this model. First, using specific antibodies, NHERF was identified in the renal proximal tubule, where it colocalized with ezrin and NHE3. NHERF2 was seen in glomeruli, the renal vasculature, and collecting duct cells, where it colocalized with ROMK. This distinct nephron localization suggests different physiologic roles for NHERF and NHERF2. Second, the signal-complex model of protein kinase A regulation of NHE3 developed in fibroblasts has been extended to epithelial cells by the development of a dominant-negative opossum kidney cell line expressing an ezrin binding domain-deficient truncation of NHERF. Preliminary studies indicate that these cells have normal basal Na+/H+ exchanger activity but a blunted inhibitory response to cAMP. Third, biochemical, biophysical, and cell experiments have indicated that NHERF binds to itself in a head-to-head configuration, raising the possibility that dimerization may alter the availability of active NHERF. The potential role of the NHERF proteins in the kidney has been expanded by recent studies indicating their involvement in the membrane targeting, trafficking, sorting, and regulation of a range of other transporters, receptors, and signaling proteins. NHERF and related PDZ-containing proteins may serve as adapters for regulation of renal transporters. PMID- 11473626 TI - Mechanisms of the inhibition of epithelial Na(+) channels by CFTR and purinergic stimulation. AB - The epithelial Na+ channel ENaC is inhibited when the cystic fibrosis transmembrane conductance regulator (CFTR) coexpressed in the same cell is activated by the cyclic adenosine monophosphate (cAMP)-dependent pathway. Regulation of ENaC by CFTR has been studied in detail in epithelial tissues from intestine and trachea and is also detected in renal cells. In the kidney, regulation of other membrane conductances might be the predominant function of CFTR. A similar inhibition of ENaC takes place when luminal purinergic receptors are activated by 5'-adenosine triphosphate (ATP) or uridine triphosphate (UTP). Because both stimulation of purinergic receptors and activation of CFTR induce a Cl(-) conductance, it is likely that Cl(-) ions control ENaC activity. PMID- 11473627 TI - Phosphorylation of a new member of the bicarbonate cotransporter superfamily. AB - Acid-base balance is regulated by cortical collecting duct cells in kidney, where the transporter functions of beta-intercalated cells are controlled by such factors as isoproterenol, cyclic adenosine monophosphate (cAMP), and the protein kinase A (PKA) pathway. The apical anion exchanger (AE) in beta-intercalated cells is thought to be involved in the secretion of bicarbonate into urine. In isolated cells, the Cl- channel was shown to be activated by isoproterenol via the PKA pathway. The importance of the PKA pathway and phosphorylation in the regulation of its transporter activity is not yet known. PMID- 11473628 TI - Liddle's syndrome: a novel mouse Nedd4 isoform regulates the activity of the epithelial Na(+) channel. AB - The epithelial Na(+) channel (ENaC), which plays an essential role in renal Na(+) handling, is composed of three subunits (alpha beta gamma), each containing a conserved PY motif at the C terminus. In Liddle's syndrome, an inherited form of salt-sensitive hypertension, the PY motifs of either beta or gamma ENaC are deleted or modified. We have recently shown that a ubiquitin-protein ligase Nedd4 binds via its WW domains to these PY motifs on ENaC, that ENaC is regulated by ubiquitination, and that Xenopus laevis Nedd4 (xNedd4) controls the cell surface pool of ENaC when coexpressed in Xenopus oocytes. Interestingly, Na(+) transporting cells, derived from mouse cortical collecting duct, express two different Nedd4 isoforms, which we have termed mNedd4-1 and mNedd4-2. Only mNedd4 2, which is orthologous to xNedd4, but not mNedd4-1, is able to regulate ENaC activity, and this property correlates with the capability to bind to the ENaC complex. Hence, Nedd4-2 may be encoded by a novel susceptibility gene for arterial hypertension. PMID- 11473629 TI - Pathophysiological mechanisms of vascular calcification in end-stage renal disease. AB - Vascular calcification has been clearly defined as a risk factor for cardiovascular mortality in the general population and is highly prevalent in end stage renal disease (ESRD), where it is associated with a number of markers of increased mortality such as left ventricular hypertrophy. The pattern of calcification in ESRD is characterized by mineral deposition in the tunica media, in contrast to non-ESRD populations, where calcification of atheromatous plaque predominates. This difference may have important clinical implications. The pathophysiological mechanisms underlying both types of vascular calcification remain to be clarified; however, current evidence suggests that they are active processes rather than passive mineral precipitation, and the presence in the vasculature of cells expressing an osteoblastic phenotype may be of central importance. In ESRD, the presence of secondary and tertiary hyperparathyroidism, disordered calcium and phosphate homeostasis, and the use of vitamin D- and calcium-based treatments in its therapy may all contribute to vascular calcification. These issues and the impact on other current and future therapies have great importance for clinical nephrology, and a better understanding of vascular calcification through a focused research effort is essential. PMID- 11473630 TI - COL4A4 mutation in thin basement membrane disease previously described in Alport syndrome. AB - BACKGROUND: Carriers of autosomal-recessive and X-linked Alport syndrome often have a thinned glomerular basement membrane (GBM) and have mutations in the COL4A3/COL4A4 and COL4A5 genes respectively. Recently, we have shown that many individuals with thin basement membrane disease (TBMD) are also from families where hematuria segregates with the COL4A3/COL4A4 locus. This study describes the first COL4A4 mutation in an individual with biopsy-proven TBMD who did not have a family member with autosomal-recessive or X-linked Alport syndrome, inherited renal failure, or deafness. METHODS: The index case and all available family members were examined for dysmorphic hematuria> 50,000/mL using phase contrast microscopy and for segregation of hematuria with the COL4A3/COL4A4 and COL4A5 loci using DNA satellite markers. COL4A4 exons from the index case were then studied using the enzyme mismatch cleavage method, and exons that demonstrated abnormal cleavage products were sequenced. RESULTS: Hematuria in this family segregated with a haplotype at the COL4A3/COL4A4 locus (P = 0.031) but not with haplotypes at the COL4A5 locus. A mutation in COL4A4 that changed C to T resulting in an arginine residue being replaced by a stop codon (R1377X) was demonstrated in exon 44, which encodes part of the alpha 4(IV) collagen sequence close to the junction with the noncollagenous domain. This mutation was present in all five family members with hematuria, but not in the four unaffected family members, 33 unrelated individuals with TBMD, or 22 nonhematuric normals. CONCLUSIONS: R1377X has been described previously in a compound heterozygous form of autosomal-recessive Alport syndrome. Our observation is evidence that TBMD can represent a carrier state for autosomal-recessive Alport syndrome in at least some individuals. PMID- 11473631 TI - Polycystin-1 transforms the cAMP growth-responsive phenotype of M-1 cells. AB - BACKGROUND: Polycystic kidney disease (PKD) is characterized by the abnormal proliferation of tubular epithelial cells. It was recently shown that the growth of PKD cyst-lining cells is stimulated by cyclic adenosine monophosphate (cAMP), whereas the growth of normal human kidney cortex cells is inhibited. METHODS: We have examined the effects of overexpressing the C-terminal cytosolic tail of mouse polycystin-1, as a membrane-targeted fusion protein, on cAMP-responsive cell proliferation in stably transfected M-1 cortical collecting duct cells. Two cell lines that express high levels of the polycystin-1 fusion protein and two control cell lines that do not express the fusion protein were tested. RESULTS: Growth of parental M-1 cells and the control cell lines was inhibited by 8-Br cAMP and by a variety of cAMP agonists. In contrast, growth of the polycystin-1 expressing clones was stimulated by cAMP. Consistent with this, the protein kinase A (PKA) inhibitor H-89 caused either a positive or a negative growth effect depending on the primary response to cAMP. PD98059 blocked the cAMP stimulation of cell proliferation, indicating that the pathway is MEK1 dependent. CONCLUSIONS: Expression of the polycystin-1 C-terminal tail disrupts normal cellular signaling and transforms the stably transfected M-1 cells to an abnormal PKD cell proliferation phenotype. PMID- 11473632 TI - Activation of renal signaling pathways in db/db mice with type 2 diabetes. AB - BACKGROUND: Altered regulation of signaling pathways may contribute to the pathogenesis of renal disease. We examined renal cortical signaling pathways in type 2 diabetes. METHODS: The status of renal cortical signaling pathways was examined in control and db/db mice with type 2 diabetes in the early phase of diabetic nephropathy associated with renal matrix expansion and albuminuria. RESULTS: Tyrosine phosphorylation of renal cortical proteins was increased in diabetic mice. Renal cortical activities of phosphatidylinositol 3-kinase (PI 3 kinase) in antiphosphotyrosine immunoprecipitates, Akt (PKB), and ERK1/2-type mitogen-activated protein (MAP) kinase activities were significantly augmented sixfold (P < 0.01), twofold (P < 0.0003), and sevenfold (P < 0.001), respectively, in diabetic mice compared with controls. A part of the increased renal cortical PI 3-kinase activity was due to insulin receptor activation, as PI 3-kinase activity associated with beta chain of the insulin receptor was increased nearly fourfold (P < 0.0235). Additionally, the kinase activity of the immunoprecipitated insulin receptor beta chain was augmented in the diabetic renal cortex, and tyrosine phosphorylation of the insulin receptor was increased. In the liver, activities of PI 3-kinase in the antiphosphotyrosine immunoprecipitates and Akt also were increased threefold (P < 0.05) and twofold (P < 0.0002), respectively. However, there was no change in the hepatic insulin receptor-associated PI 3-kinase activity. Additionally, the hepatic ERK1/2-type MAP kinase activity was inhibited by nearly 50% (P < 0.01). CONCLUSIONS: These studies demonstrate that a variety of receptor signaling pathways are activated in the renal cortex of mice with type 2 diabetes, and suggest a role for augmented insulin receptor activity in nephropathy of type 2 diabetes. PMID- 11473633 TI - A novel somatostatin analogue prevents early renal complications in the nonobese diabetic mouse. AB - BACKGROUND: PTR-3173 (S) is a novel somatostatin analogue that has been found to exert a prolonged inhibitory action on the growth hormone (GH)-insulin-like growth factor (IGF)-I axis, but not on insulin secretion. We investigated the potential effect of this agent on the development of markers of diabetic nephropathy in the nonobese diabetic (NOD) mouse model of insulin-dependent diabetes. METHODS: Female diabetic NOD mice treated with PTR-3173 (DS group) or saline (D) and their control groups of nonhyperglycemic age-matched littermates (C) and C mice treated with PTR-3173 (CS) were sacrificed three weeks after onset of diabetes. RESULTS: Serum GH was elevated in the D group, decreased in the DS group, and unchanged in the CS group. Serum IGF-I was significantly decreased in both the D and DS groups. Kidney weight, glomerular volume, albuminuria, and creatinine clearance were increased in the D animals and showed a trend toward normalization in the DS animals. Renal extractable IGF-I protein and IGFBP1 mRNA were increased in the D group and normalized in the DS group. CONCLUSIONS: GH antagonism by PTR-3173 has a blunting effect on renal/glomerular hypertrophy, albuminuria, and glomerular filtration rate (GFR) in diabetic NOD mice. This phenomenon is apparently associated with the prevention of renal IGF-I accumulation. Thus, modulation of GH effects may have beneficial therapeutic implications in diabetic nephropathy. PMID- 11473634 TI - Intraperitoneal atrial natriuretic peptide increases peritoneal fluid and solute removal. AB - BACKGROUND: Atrial natriuretic peptide (ANP) is a hormone with well-known diuretic and vasodilating properties. Recently it was reported that ANP could increase the peritoneal fluid formation and increase peritoneal solute clearance. This study investigated the effect of ANP on peritoneal fluid and solute transport characteristics. METHODS: Eighteen male Sprague-Dawley rats were divided into three groups. A four-hour dwell study using 25 mL 2.27% glucose dialysis solution with 50 microg/kg ANP (N = 6, H-ANP) or 5 microg/kg ANP (N = 6, L-ANP) or without ANP (N = 8, control) and frequent dialysate and blood sampling was done in each rat. Radiolabeled human albumin (RISA) was added to the solution as an intraperitoneal volume marker. RESULTS: The intraperitoneal volume was significantly higher in the H-ANP group as compared with the control group and the L-ANP group. The drainage volume was 26.2 +/- 1.1, 25.5 +/- 0.7, and 29.8 +/- 0.8 mL for the control, L-ANP, and H-ANP groups, respectively (P < 0.01). This was related to significant differences in the peritoneal fluid absorption rates (K(E); estimated as the RISA elimination coefficient): 39 +/- 3, 38 +/- 3, and 19 +/- 4 microL/min, and in the direct lymphatic absorption rate (K(EB); estimated as the clearance of RISA from dialysate to blood): 7 +/- 1, 6 +/- 1, and 4 +/- 1 microL/min for the control, L-ANP, and H-ANP groups, respectively (all P < 0.01). No differences were found in the intraperitoneal volume, K(E), and K(EB) between the control group and the L-ANP group. The diffusive mass transport coefficient (K(BD)) for urea, sodium, potassium, and total protein did not differ among the three groups. However, the glucose D/D(0) was significantly higher, and the K(BD) for glucose was significantly lower in the H-ANP group as compared with the other two groups. Solute clearances (+175% for sodium and +26% for potassium) were significantly increased in the H-ANP group, mainly as a result of the increased fluid removal in this group. CONCLUSIONS: Our results suggest that ANP may decrease peritoneal fluid absorption (by 51%, partially because of decreasing the direct lymphatic absorption), resulting in a significant increase in peritoneal fluid removal and small solute clearances. While the basic diffusive permeability of the peritoneal membrane was not changed, the peritoneal glucose absorption was retarded by adding ANP to peritoneal dialysate, perhaps through interaction of ANP with glucose metabolism. PMID- 11473635 TI - Lipoprotein-X stimulates monocyte chemoattractant protein-1 expression in mesangial cells via nuclear factor-kappa B. AB - BACKGROUND: Lipoprotein-X (Lp-X) is an abnormal lipoprotein found in the plasma of patients with familial lecithin:cholesterol acyltransferase (LCAT) deficiency. The majority of patients with this disorder develop progressive glomerulosclerosis. One key event in the pathogenesis of glomerulosclerosis is the infiltration of monocytes into affected glomeruli. Mesangial cells can synthesize and secrete monocyte chemoattractant protein-1 (MCP-1), an important chemoattractant for monocytes. The objective of the present study was to examine the effect of Lp-X on MCP-1 expression in mesangial cells leading to an enhanced monocyte chemotaxis and to elucidate the mechanisms involved in this process. METHODS: Lp-X was isolated from the plasma of a patient with familial LCAT deficiency. After rat mesangial cells were incubated with Lp-X for four or six hours, the expression of MCP-1 mRNA was determined by nuclease protection assay, and MCP-1 protein was measured by Western immunoblotting analysis. Monocyte chemotaxis was determined by using a Micro Chemotaxis Chamber. RESULTS: Lp-X (50 to 100 nmol/mL) stimulated mesangial cell MCP-1 mRNA expression (137 to 220%) and MCP-1 protein levels (233 to 375%). Conditioned media collected from Lp-X-treated mesangial cells stimulated human acute monocytic leukemia (THP-1) monocyte chemotaxis (165 to 200%). The increase in MCP-1 expression in mesangial cells was associated with an elevation of intracellular diacylglycerol levels, and activation of protein kinase C (PKC) as well as nuclear factor-kappa B (NF-kappa B). CONCLUSION: These results suggest that Lp-X participates in the pathogenesis of glomerulosclerosis and subsequent renal failure in familial LCAT deficient patients by stimulating monocyte infiltration via a mechanism involving mesangial MCP-1 expression. PMID- 11473636 TI - Transforming growth factor-beta suppresses macrophage-induced mesangial cell fibronectin expression. AB - BACKGROUND: We have previously shown that macrophages are able to promote prosclerotic responses in rat mesangial cells. Th2-type cytokines, including interleukin-10 (IL-10), IL-13, and IL-4 as well as transforming growth factor beta (TGF-beta), are known to have suppressive effects on various aspects of macrophage function. In the current study, we investigated the effect of TGF-beta pretreatment on the ability of macrophages to induce fibronectin expression. RESULTS: Conditioned medium from TGF-beta pretreated macrophages (MPCM(TGF)) induced lower fibronectin levels in mesangial cells in both the secreted and cell associated forms, compared with conditioned medium from standard macrophages (MPCM) (5.5 +/- 0.2 vs. 3.4 +/- 0.3 and 4.05 +/- 0.45 vs. 2.3 +/- 0.2-fold increase over medium alone for MPCM versus MPCM(TGF) in supernatants and cell lysates, respectively). Northern blot analysis demonstrated that fibronectin message was marginally reduced to 0.88 +/- 0.04 (P < 0.03 vs. MPCM, N = 3) of MPCM-induced levels. However, mesangial cell transin mRNA levels induced in response to MPCM(TGF) were 2.29 +/- 0.47-fold greater than those induced by standard MPCM (P = 0.03 vs. MPCM, N = 4). TIMP-1 mRNA levels were also increased in response to MPCM(TGF), but only by 1.43 +/- 0.1-fold (P = 0.02 vs. MPCM, N = 5). Casein-FITC digestion studies confirmed that MPCM(TGF) stimulated more mesangial cell caseinolytic activity than did MPCM. In addition, MPCM-mediated up regulation of mesangial cell TGF-beta mRNA and protein expression was significantly reduced in response to conditioned medium from macrophages pretreated with TGF-beta. CONCLUSION: This study suggests that TGF-beta is able to regulate negatively the profibrotic effects of macrophages on mesangial cells by both enhancing matrix degradation and reducing synthesis. PMID- 11473637 TI - p38 MAPK and MAPK kinase 3/6 mRNA and activities are increased in early diabetic glomeruli. AB - BACKGROUND: The p38 mitogen-activated protein kinase (MAPK) pathway is activated by several stress factors, potentially leading to cellular apoptosis and growth. Little is known about the pattern of glomerular p38 MAPK pathway activation during the course of diabetic nephropathy (DN). We examined the activity and expression of the p38 MAPK pathway members, p38 MAPK, MKK3/6, cAMP-responsive element binding protein (CREB), and MAPK phosphatase-1 (MKP-1), in experimental DN in rats over the course of four months. METHODS: Control (C; N = 16) and diabetic (DM; N = 16) rats were studied. Four rats from each group were sacrificed monthly, and competitive reverse transcription-polymerase chain reaction and Western blot were performed with microdissected and sieved glomeruli, respectively. RESULTS: Glomerular p38 MAPK mRNA expression was significantly higher in DM than C (P < 0.01) throughout the four-month period. Western blot revealed an average 3.1-fold increase in p38 MAPK protein throughout the study period (P < 0.05). However, p38 MAPK activity was significantly increased only in one- and two-month diabetic glomeruli. Glomerular MKK3/6 and CREB mRNA as well as activity were significantly increased only in one- and two month DM compared with C. MKP-1 mRNA showed a similar pattern. CONCLUSIONS: Glomerular p38 MAPK activity was increased in early DN. Parallel to this, we also showed, to our knowledge for the first time, that there were increased MKK3/6 and CREB activities and mRNA expression. This activated p38 MAPK pathway in diabetic glomeruli may, in part, play a role in the pathogenesis of early hypertrophy and extracellular matrix accumulation. PMID- 11473638 TI - Hyperosmolality activates Akt and regulates apoptosis in renal tubular cells. AB - BACKGROUND: The novel serine-threonine kinase Akt is a critical enzyme in cell survival. We investigated the roles of the Akt pathway and apoptotic signals in (1) Madin-Darby canine kidney (MDCK) cells in a hyperosmotic condition in vitro and (2) in the inner medulla of dehydrated rat in vivo. METHODS: The in vivo experiments were performed in 24- and 48-hour water-restricted rats. Hyperosmolality-stimulated Akt phosphorylation was examined in MDCK cells. Phosphatidylinositol 3-kinase (PI3-K) inhibitors, the dominant-negative mutant of PI3-K, the dominant-negative mutant of Akt, and the dominant-active form of Akt were used to examine the roles of the PI3-K/Akt pathways in renal tubular cell apoptosis. RESULTS: The amount of phosphorylated Akt protein was increased in the inner medulla of dehydrated rats. Hyperosmolality induced by the addition of NaCl, urea, and raffinose phosphorylated Akt in MDCK cells in an osmolality dependent manner. PI3-K inhibitors and the dominant-negative mutant of PI3-K inhibited the hyperosmolality-induced phosphorylation of Akt. Raising the media osmolality from a normal level to 500 or 600 mOsm/kg H2O final osmolality elicited apoptotic changes such as nucleosomal laddering of DNA and an increment of caspase-3 activity and increased activity in the cell death enzyme-linked immunosorbent assay. Dominant-active Akt prevented the mild hyperosmolality induced apoptosis, while inhibition of the PI3-K/Akt pathways promoted apoptosis. CONCLUSION: The Akt pathway is activated by hyperosmolality in vitro and in vivo, and activation of Akt prevents the mild hyperosmolality-induced apoptotic changes in MDCK cells. PI3-K/Akt pathways are involved in a hypertonic condition that confers the balance between cell survival and apoptosis. PMID- 11473639 TI - Morphogenesis of MDCK cells in a collagen gel matrix culture under stromal adipocyte-epithelial cell interaction. AB - BACKGROUND: The stromal-epithelial cell interaction is essential for epithelial morphogenesis. Recently, the specific stromal cell type adipocytes, which abundantly exist in perirenal adipose tissue, have been suggested to affect the biological behavior of some epithelial cell types. However, adipocyte-renal epithelial cell interaction remains unclear. We thus examined the effects of adipocytes on the morphogenesis of renal epithelial cells. METHODS: The renal epithelial cell line, Madin-Darby canine kidney (MDCK), cells were cultured in three-dimensional collagen gel matrix with or without mature unilocular adipocytes. Cultures cells were examined by histochemistry, immunohistochemistry, and electron microscopy. RESULTS: Adipocytes extensively promoted the tubule formation of MDCK cells in two different manners. In the first type, after approximately 20% of MDCK cells actively adhered to adipocytes; they organized double-cell structured tubules between the adipocytes and the gel, contacting directly with the entire surface of the adipocytes. In the second type, approximately 70% of MDCK cells apart from adipocytes also formed tubules that had no contact with adipocytes. The component cells of both tubule types at the apical side showed microvilli and peanut agglutinin lectin-positive stain. These cells at the basal side had the basal lamina and type IV collagen-positive stain. CONCLUSIONS: These results indicate that the specific stromal cell type adipocytes cause MDCK cells to organize the well-polarized tubular structures in two different manners according to their direct and indirect interactions, suggesting that adipocytes may be involved in the regulatory mechanism of renal epithelial morphogenesis. PMID- 11473640 TI - Expression, regulation, and function of inhibitor of apoptosis family genes in rat mesangial cells. AB - BACKGROUND: The inhibitor of apoptosis (IAP) family of proteins regulates programmed cell death triggered by various stimuli. The purpose of this investigation was to examine the expression, regulation, and function of IAP genes in cultured rat mesangial cells. METHODS: Basal and inducible expression of c-IAP1, c-IAP2, XIAP, and TIAP mRNAs was examined in mesangial cells, isolated glomeruli, and other cell lines under unstimulated and tumor necrosis factor alpha (TNF-alpha)-stimulated conditions. To examine a role of nuclear factor kappa B (NF-kappa B) in the regulation of IAPs, expression of IAPs in NF-kappa B inactive mesangial cells was compared with that in wild-type cells. To investigate roles of IAPs in mesangial cell apoptosis, NF-kappa B--inactive cells were stably supertransfected with c-IAP1 or c-IAP2, and the susceptibility of these cells to TNF-alpha--induced apoptosis was evaluated quantitatively. RESULTS: Substantial, constitutive expression of c-IAP2, XIAP, and TIAP was observed in serum-deprived rat mesangial cells and c-IAP2 and XIAP in isolated normal rat glomeruli. In response to TNF-alpha, expression of c-IAP1 and c-IAP2 was induced in HeLa cells and ECV304 endothelial cells, but not in mesangial cells. In contrast to previous reports on other cell types, the expression of IAPs in rat mesangial cells was independent of NF-kappa B; that is, expression levels of IAPs in NF-kappa B--inactive cells were same as those in NF-kappa B- active cells under both unstimulated and TNF-alpha--stimulated conditions. Even without the induction of IAPs, NF-kappa B--active mesangial cells were more resistant to TNF-alpha--induced apoptosis than NF-kappa B--inactive cells. Interestingly, overexpression of either c-IAP1 or c-IAP2 completely compensated for the lack of resistance to apoptosis in NF-kappa B--inactive cells. CONCLUSIONS: IAPs are constitutively expressed in cultured rat mesangial cells and isolated normal rat glomeruli. IAPs can contribute to the survival of rat mesangial cells, but unexpectedly, these molecules are not involved in the TNF alpha--induced, NF-kappa B--dependent cytoprotection in this cell type. PMID- 11473641 TI - PAI-1 deficiency attenuates the fibrogenic response to ureteral obstruction. AB - BACKGROUND: Progressive renal disease is characterized by the induction of plasminogen activator inhibitor-1 (PAI-1), suggesting that impaired activity of the renal plasmin cascade may play a role in renal fibrosis. METHODS: To test this hypothesis, the severity of renal fibrosis caused by unilateral ureteral obstruction (UUO) was compared in PAI-1 wild-type (+/+) and PAI-1 deficient (-/-) mice. The extent of interstitial inflammation and fibrosis, renal plasminogen activator and plasmin activity, and renal expression of profibrotic genes was evaluated after 3, 7, and 14 days of UUO. RESULTS: Renal PAI-1 mRNA levels increased 8- to 16-fold in the +/+ mice after UUO surgery, and PAI-1 protein was detected in kidney homogenates. Interstitial fibrosis was significantly attenuated in -/- mice compared with +/+ mice at day 7 and day 14, based on the interstitial area stained with picrosirius red and total kidney collagen content. However, neither the mean renal plasminogen activator nor plasmin activities were increased in -/- mice compared with +/+ mice. The number of interstitial macrophages were significantly lower in the -/- mice three and seven days after UUO; interstitial myofibroblasts were significantly fewer at three days. At the same time points, this altered interstitial cellularity was associated with a significant reduction in renal mRNA levels for transforming growth factor-beta and procollagens alpha 1(I) and alpha 1(III). CONCLUSIONS: These studies establish an important fibrogenic role for PAI-1 in the renal fibrogenic response. The results demonstrate that one important fibrosis-promoting function of PAI-1 is its role in the recruitment of fibrosis-inducing cells, including myofibroblasts and macrophages. PMID- 11473642 TI - Renal fibroblast-like cells in Goodpasture syndrome rats. AB - BACKGROUND: The extent of renal fibrosis is the best predictor for functional outcomes in a variety of progressive renal diseases. Interstitial fibroblast-like cells (FbLCs) are presumably involved in the fibrotic process. However, such FbLCs have never been well characterized in the kidney. METHODS: We characterized renal FbLCs in the nephritic kidney (in which the number of FbLCs and extracellular matrix accumulation were significantly increased) with regards to their expression of phenotypic and functional markers using day 49 Goodpasture syndrome (GPS) rats. RESULTS: Within the renal cortical interstitium, there were a number of alpha-smooth muscle actin(+) (alpha-SMA(+)) FbLCs, negative for vimentin (VIM) and transforming growth factor-beta 1, and not equipped with well developed rough endoplasmic reticulum and actin-stress fibers. All of these findings were incompatible with the typical features of granulation tissue alpha SMA(+) myofibroblasts. On the other hand, FbLCs negative for alpha-SMA and VIM produced alpha1(I) procollagen in the nephritic kidney. CONCLUSION: A number of FbLC populations reside within the cortical interstitium of the kidney in GPS rats, each of which is likely to have developed independently in response to the local conditions of the nephritic kidney, contributing to renal fibrogenesis. Further studies are needed to clarify the key type of FbLC that orchestrates other members to produce renal fibrosis. PMID- 11473643 TI - Compensated heart failure predisposes to outer medullary tubular injury: studies in rats. AB - BACKGROUND: Heart failure (HF) is considered a putative factor predisposing to acute renal failure (ARF). Since outer medullary hypoxic injury may play an important role in the pathogenesis of acute tubular necrosis, we explored the impact of experimental HF on the propensity to develop ARF with hypoxic medullary injury following the inhibition of prostaglandin and nitric oxide synthesis. METHODS: Compensated, high-output HF was induced in Sprague-Dawley rats by aorto caval fistula. At the eighth to ninth postoperative day, the rats were injected with indomethacin and N(omega) nitro-L-arginine methyl ester (L-NAME; ARF protocol) and were sacrificed 24 hours later for morphologic evaluation. RESULTS: Kidney function comparably declined in HF-ARF rats and in control sham operated animals (CTR-ARF). Nevertheless, outer medullary hypoxic damage with medullary thick ascending limb (mTAL) necrosis occurred almost exclusively in the HF-ARF group (11 +/- 4% vs. 0.2 +/- 0.2% of tubules in CTR-ARF, P < 0.03). In a third group of HF animals subjected to vehicles only (HF-Nil), kidney function was preserved and renal morphology remained intact. Papillary-tip necrosis was consistently found in all animals subjected to indomethacin and L-NAME, irrespective of preconditioning. Morphometric evaluation disclosed that HF was not associated with mTAL hypertrophy. CONCLUSIONS: Incipient HF predisposes to hypoxic outer medullary injury, probably reflecting the impact of regional vasoconstrictive stimuli rather than tubular hypertrophy when protective local vasodilating mechanisms are hampered. The presence and extent of outer medullary hypoxic damage cannot be predicted from the functional derangement, which in the experimental settings may also represent prerenal azotemia or papillary damage. PMID- 11473644 TI - Tubules are the major site of M-CSF production in experimental kidney disease: correlation with local macrophage proliferation. AB - BACKGROUND: Local proliferation of macrophages occurs within both the glomerulus and the interstitium in severe forms of human and experimental glomerulonephritis and plays an important role in amplifying renal injury. Macrophage colony stimulating factor (M-CSF) is thought to be the growth factor driving this local macrophage proliferation. Previous studies have found that glomeruli are the predominant source of M-CSF production. However, this is difficult to reconcile with the prominent macrophage accumulation and proliferation seen in the interstitial compartment in glomerulonephritis. To address this issue, we localized M-CSF expression in rat models of glomerular versus tubulointerstitial injury and examined its relationship to local macrophage proliferation. METHODS: M-CSF expression (Northern blotting, in situ hybridization, immunostaining, Western blotting) and local macrophage proliferation (double immunostaining) was examined in normal rat kidney on days 1 and 14 of rat anti-glomerular basement membrane (anti-GBM) glomerulonephritis and on day 5 following unilateral ureteric obstruction. RESULTS: M-CSF mRNA and protein expression were identified in small numbers of glomerular podocytes, approximately 25% of cortical tubules, and most medullary tubules in normal rat kidney. Northern blotting showed a significant increase in whole kidney M-CSF mRNA in rat anti-GBM glomerulonephritis. Up regulation of glomerular and, most prominently, tubular M-CSF production was confirmed by three independent methods: in situ hybridization, immunostaining, and Western blotting. The increase in M-CSF expression colocalized with local macrophage proliferation (ED1+PCNA+ cells) in both the glomerulus and tubulointerstitium. On day 5 after ureter ligation, there was a significant increase in tubular M-CSF mRNA and protein expression in the obstructed kidney, with no change in glomerular M-CSF. In parallel with M-CSF expression, macrophage accumulation and proliferation was prominent in the interstitium, but was absent from glomeruli. CONCLUSIONS: The tubular epithelial cell is the major site of M CSF production within the injured kidney. Indeed, substantial macrophage accumulation and local proliferation can occur in the tubulointerstitium in the absence of glomerular inflammation. These results suggest that M-CSF production within the kidney, particularly by tubular epithelial cells, plays an important role in regulating local macrophage proliferation in experimental kidney disease. PMID- 11473645 TI - Podocyte-specific expression of a novel trans-Golgi protein Vear in human kidney. AB - BACKGROUND: Vear is a recently identified Golgi apparatus-associated protein. It has been suggested to be involved in vesicular trafficking between the Golgi and the vacuolar/lysosomal system. Proteins similar to Vear have also been shown to interact with activated ARF proteins (ADP ribosylation factor), and they are probably involved in membrane trafficking from the trans-Golgi network (TGN). We have previously shown that Vear is widely distributed in human tissues, with an especially high level of mRNA in the kidney. This study further characterizes the distribution and subcellular localization of Vear in normal adult kidney and shows its association with glomerulogenesis in fetal kidney. METHODS: Immunofluorescence and immunoelectron microscopy were used to study the expression of Vear in fetal and adult kidney. The expression of Vear in isolated glomeruli was shown by immunoblotting. The distribution of its mRNA was analyzed by using in situ and Northern hybridization. RESULTS: In situ hybridization and immunofluorescence microscopy showed that in the kidney, Vear is present in glomerular structures. By fluorescence microscopy, the immunoreactivity for Vear was found only in podocytes, as judged by its distinct colocalization with podocalyxin and vimentin, well-established marker proteins of podocytes. Its specific expression in the glomeruli versus other compartments of the kidney was also verified by Western blotting. By using immunogold electron microscopy, Vear was seen in the Golgi apparatus, tubulovesicular structures, and membranes adjacent to the Golgi complex. In fetal kidney, expression of Vear coincided with the formation of segmental structures of the glomeruli. It was first seen close to the undifferentiated luminal cells at the vesicular stage and increasingly in the differentiating podocytes at the more advanced stages of glomerulogenesis. CONCLUSIONS: In the kidney, Vear shows a distinct, specific, and developmentally regulated expression in glomerular podocytes. This suggests that Vear has a specific function in podocytes. It could be associated with the known high secretory and synthetic activity of the podocytes, especially the production of the basement membrane components, which are critically involved in the glomerulogenesis and the maintenance of the glomerular function. PMID- 11473646 TI - Osteopontin expression in human cyclosporine toxicity. AB - BACKGROUND: Osteopontin is a secreted phosphoprotein that has a number of diverse biological functions, including cell signaling, mediation of cell adhesion, migration, and chemoattraction of monocytes/macrophages. Up-regulation of osteopontin expression by proximal tubular epithelium has been demonstrated in both human and rodent models of renal injury in association with macrophage influx. METHODS: We studied the expression of osteopontin protein and mRNA in renal donor biopsies (N = 7) and renal transplant biopsies with cyclosporine A toxicity (N = 23) by immunohistochemistry and in situ hybridization. Serial tissue sections were immunostained with a monocyte/macrophage marker, CD68, to demonstrate the pattern of macrophage infiltration. RESULTS: Strong osteopontin expression was observed in the majority of pretransplant donor biopsies in the absence of any macrophage infiltration. In the biopsies with cyclosporine toxicity, osteopontin expression was widespread and demonstrated moderate immunohistochemical signal intensity that did not correlate with the number of interstitial macrophages present. CONCLUSIONS: Strong osteopontin protein and mRNA expression by tubular epithelium was observed in pretransplant donor biopsies and in biopsies with cyclosporine toxicity without an inflammatory cell infiltration. Therefore, osteopontin expression alone is insufficient to serve as the principal mediator of intrarenal monocyte/macrophage influx in the transplant setting. PMID- 11473647 TI - Cloning of rodent megsin revealed its up-regulation in mesangioproliferative nephritis. AB - BACKGROUND: We recently cloned a new human mesangium-predominant gene, megsin. Megsin is a novel member of the serine protease inhibitor (serpin) superfamily. To elucidate functional roles of this gene, we cloned megsin in rodents and investigated its role in a rat nephritis model. METHODS: Megsin homologues were cloned from cultured rat and mouse mesangial cDNAs utilizing polymerase chain reaction (PCR) with degenerative primers. Expression of megsin in three different types of resident glomerular cells was investigated by PCR. Levels of megsin mRNA expression at various time points in the anti-Thy1 rat nephritis model were studied by semiquantitative PCR and Northern blotting analysis. In order to investigate megsin protein expression in anti-Thy1 nephritis rats, we raised antibody against rat megsin-specific synthetic peptide, with which immunohistochemical studies were performed. RESULTS: Rat and mouse megsins were composed of 380 amino acids, which revealed 75.3 and 73.9% identity, respectively, with human megsin at the amino acid level. Characteristic features as an inhibitory serpin were conserved in both rat and megsin megsins. PCR analysis revealed expression of megsin in cultured mesangial cells but not in glomerular epithelial or endothelial cells. In anti-Thy1 nephritis rats, semiquantitative PCR and Northern blotting showed that expression of megsin mRNA was up-regulated in glomeruli at day 8. Immunohistochemical studies demonstrated the prominent accumulation of megsin in glomeruli at the same time point. Megsin was mainly localized in mesangial area. The megsin expression level returned to the basal level at day 28. CONCLUSION: Sequences of megsin were well conserved among different species. Rat megsin was also predominantly expressed in mesangial cells. Expression of megsin was up-regulated at the peak of hypercellularity and matrix accumulation in the mesangioproliferative glomerulonephritis model, suggesting that megsin may participate in the process of glomerulosclerosis by modulating extracellular matrix deposition or cell survival. PMID- 11473648 TI - Mycophenolate mofetil combined with a cyclooxygenase-2 inhibitor ameliorates murine lupus nephritis. AB - BACKGROUND: Approaches to the treatment of lupus nephritis include immunosuppressants associated with anti-inflammatory drugs, mainly steroids, which, however, cause major side effects. Mycophenolate mofetil (MMF) has been described as being less toxic than conventional immunosuppressants, and it was effective in preventing progressive nephritis in lupus mice. Our study evaluated the therapeutic effect of MMF in NZB/W F1 hybrid mice with established disease. We also examined the combination of MMF with a selective cyclooxygenase-2 (COX-2) inhibitor, DFU, based on previous findings of excessive renal production of COX-2 derived thromboxane A2 (TXA2) in lupus nephritis. METHODS: Four groups of NZB/W mice (N = 30 each group), starting at five months of age, were given daily by gavage the following: vehicle, MMF 60 mg/kg, DFU 3 mg/kg, or MMF + DFU. Fifteen mice for each group were used for the survival studies, and the remaining mice were sacrificed at nine months. RESULTS: MMF or DFU alone partially delayed the onset of proteinuria compared with vehicle. Combined therapy was significantly (P < 0.05) more effective than single drugs. Animal survival was partially ameliorated by MMF and significantly improved by the drug combination in comparison with the vehicle (P = 0.005) and DFU alone (P < 0.03). At nine months, serum blood urea nitrogen (BUN) levels were lower in all of the treated groups than in the vehicle group. Renal damage was also limited, but to a greater extent in mice given the combined therapy. In untreated mice, renal COX-2 mRNA expression was up-regulated, and generation of TXB(2), the stable breakdown product of TXA(2), increased. DFU prevented the abnormal renal TXB(2) production, confirming the COX-2 origin of this eicosanoid, whereas renal 6-keto-PGF(1 alpha) and prostaglandin E(2) (PGE(2)) were not affected substantially. CONCLUSIONS: These results offer a strong case for exploring the possibility that in humans MMF combined with COX-2 inhibitors has a role in the treatment options for lupus nephritis. This combined drug therapy may be at least as effective as steroids but without the obvious nephrotoxicity of the latter. PMID- 11473649 TI - Kidney protection against autoreactive CD8(+) T cells distinct from immunoprivilege and sequestration. AB - BACKGROUND: The kidney tubulointerstitium has been reported to be protected from T-cell--mediated damage by sequestration from the T-cell compartment. We examined the ability of autoreactive T cells to infiltrate the kidney in a transgenic mouse model. METHODS: RIP-mOVA transgenic mice express the model autoantigen, membrane-bound ovalbumin (mOVA), in kidney proximal tubular cells and pancreatic beta cells. OVA-specific CD8(+) T cells (OT-I cells) were transferred into these recipient mice and their immune response against pancreas and kidney tissue was compared. RESULTS: When OVA-specific CD8(+) T cells (OT-I cells) were injected into RIP-mOVA mice, they were activated in the renal and pancreatic lymph nodes by cross-presentation. These in vivo-activated OT-I cells caused the destruction of pancreatic islets leading to autoimmune diabetes, but did not infiltrate the kidney. Neither CD95--CD95 ligand interactions, which have been proposed to induce apoptosis in T cells infiltrating immunologically privileged sites, nor CD30 signaling was responsible for the lack of kidney infiltration. When OT-I cells were activated in vitro prior to injection, they could infiltrate the kidney and caused acute renal failure when injected in high numbers. CONCLUSIONS: A mechanism distinct from previously described organ-specific protective mechanisms such as sequestration of antigen or CD95-mediated immunoprivilege contributes to the protection of the kidney tubulointerstitium from infiltration by autoreactive CD8(+) T cell. PMID- 11473650 TI - Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft. AB - BACKGROUND: The underlying mechanisms of renal transplant dysfunction are poorly understood. There is little information on tubular function in kidney grafts. The cDNAs encoding kidney-specific cell surface proteins required for renal reabsorption of sodium (sodium cotransporter in thick ascending limb of Henle, rBSC1) and water (apical water channel in collecting duct, AQP2) have been recently identified. Since transcripts of these proteins are up-regulated in dehydration in association with maximal concentration of urine, we examined urinary concentrating ability and expression levels of mRNA of these proteins in kidney isografts. METHODS: Male Sprague-Dawley rats underwent syngeneic renal transplantation or unilateral nephrectomy (UNX) and were deprived of water for 24 hours at six weeks after the operation when histological and functional compensation of the intact kidney was complete. Blood and urinary samples were collected before and after dehydration. The amount of rBSC1 or AQP2 mRNA was measured using competitive polymerase chain reaction (PCR) by inducing a point mutation at the middle of PCR product for rBSC1 or by deleting 180 bp from 780 bp PCR product for AQP2, respectively. The protein expression was examined by Western blot analysis. RESULTS: Both groups of rats demonstrated the same levels of compensatory renal hypertrophy (approximately 60% weight increase) and plasma creatinine values. Histological examination revealed enlarged glomeruli and tubules, but no findings of ischemic damage, such as tubular atrophy or interstitial changes. Urinary concentration was noted in the UNX rats but not in rats with kidney grafts. Competitive PCR demonstrated that dehydration did not increase rBSC1 and AQP2 transcripts in rats with kidney transplantation. Immunoblot analysis confirmed that the marked increase of both rBSC1 and AQP2 proteins was noted only in the remnant kidney of dehydrated rats. CONCLUSIONS: Rats with kidney isografts have a limited capacity to concentrate urine and, at the same time, fail to increase rBSC1 and AQP2 transcripts. This suggests that there is a prolonged damage of renal tubules by ischemia or denervation of the donor kidney, both of which are inevitable in the transplantation procedure. PMID- 11473651 TI - "Avian-type" renal medullary tubule organization causes immaturity of urine concentrating ability in neonates. AB - BACKGROUND: While neonatal kidneys are not powerful in concentrating urine, they already dilute urine as efficiently as adult kidneys. To elucidate the basis for this paradoxical immaturity in urine-concentrating ability, we investigated the function of Henle's loop and collecting ducts (IMCDs) in the inner medulla of neonatal rat kidneys. METHODS: Analyses of individual renal tubules in the inner medulla of neonatal and adult rat kidneys were performed by measuring mRNA expression of membrane transporters, transepithelial voltages, and isotopic water and ion fluxes. Immunofluorescent identification of the rCCC2 and rCLC-K1 using polyclonal antibodies was also performed in neonatal and adult kidney slices. RESULTS: On day 1, the transepithelial voltages (V(Ts)) in the thin ascending limbs (tALs) and IMCDs were 14.6 +/- 1.1 mV (N = 27) and -42.7 +/- 6.1 mV (N = 14), respectively. The V(Ts) in the thin descending limbs (tDLs) were zero on day 1. The V(Ts) in the tALs were strongly inhibited by luminal bumetanide or basolateral ouabain, suggesting the presence of a NaCl reabsorption mechanism similar to that in the thick ascending limb (TAL). The diffusional voltage (V(D)) of the tAL due to transepithelial NaCl gradient was almost insensitive to a chloride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB). The V(Ts) in the IMCDs were strongly inhibited by luminal amiloride. On day 1, both the tDL and tAL were impermeable to water, indicating the water impermeability of the entire loop. Diffusional water permeability (P(dw)) and urea permeabilities (P(urea)) in the IMCDs indicated virtual impermeability to water and urea on day 1. Stimulation by vasopressin (1 nmol/L) revealed that only P(dw) was sensitive to vasopressin by day 14. A partial isoosmolar replacement of luminal urea by NaCl evoked negligible water flux across the neonatal IMCDs, indicating the absence of urea-dependent volume flux in the neonatal IMCD. These transport characteristics in each neonatal tubule are similar to those in quail kidneys. Identification of mRNAs and immunofluorescent studies for specific transporters, including rAQP-1, rCCC2, rCLC-K1, rENaC beta subunit, rAQP-2, and rUT-A1, supported these findings. CONCLUSION: We hypothesize that the renal medullary tubule organization of neonatal rats shares a tremendous similarity with avian renal medulla. The qualitative changes in the organization of medullary tubules may be primarily responsible for the immature urine-concentrating ability in mammalian neonates. PMID- 11473652 TI - Glycosphingolipids modulate renal phosphate transport in potassium deficiency. AB - BACKGROUND: Potassium (K) deficiency (KD) and/or hypokalemia have been associated with disturbances of phosphate metabolism. The purpose of the present study was to determine the cellular mechanisms that mediate the impairment of renal proximal tubular Na/Pi cotransport in a model of K deficiency in the rat. METHODS: K deficiency in the rat was achieved by feeding rats a K-deficient diet for seven days, which resulted in a marked decrease in serum and tissue K content. RESULTS: K deficiency resulted in a marked increase in urinary Pi excretion and a decrease in the V(max) of brush-border membrane (BBM) Na/Pi cotransport activity (1943 +/- 95 in control vs. 1184 +/- 99 pmol/5 sec/mg BBM protein in K deficiency, P < 0.02). Surprisingly, the decrease in Na/Pi cotransport activity was associated with increases in the abundance of type I (NaPi-1), and type II (NaPi-2) and type III (Glvr-1) Na/Pi protein. The decrease in Na/Pi transport was associated with significant alterations in BBM lipid composition, including increases in sphingomyelin, glucosylceramide, and ganglioside GM3 content and a decrease in BBM lipid fluidity. Inhibition of glucosylceramide synthesis resulted in increases in BBM Na/Pi cotransport activity in control and K-deficient rats. The resultant Na/Pi cotransport activity in K-deficient rats was the same as in control rats (1148 +/- 52 in control + PDMP vs. 1152 +/- 61 pmol/5 sec/mg BBM protein in K deficiency + PDMP). These changes in transport activity occurred independent of further changes in BBM NaPi-2 protein or renal cortical NaPi-2 mRNA abundance. CONCLUSION: K deficiency in the rat causes inhibition of renal Na/Pi cotransport activity by post-translational mechanisms that are mediated in part through alterations in glucosylceramide content and membrane lipid dynamics. PMID- 11473653 TI - Kidney-specific chromosome transfer in genetic hypertension: the Dahl hypothesis revisited. AB - BACKGROUND: A central dogma in the field of essential hypertension research is that the genetic transmission of increased blood pressure is determined solely by the genotype of the kidney. This concept is based in large part on studies in experimental rat models of spontaneous hypertension in which transplantation of a kidney from a hypertensive strain into a normotensive strain was reported to increase blood pressure, and transplantation of a kidney from a normotensive strain into a hypertensive strain was reported to decrease blood pressure. The enduring interpretation of these now classic experiments remains virtually unchanged from the view originally espoused a quarter century ago by Lewis Dahl, one of the founding fathers of the field of genetic hypertension research: "Blood pressure is determined by the genotype of the donor kidney and not the genotype of the recipient." METHODS: To test the Dahl hypothesis, we determined the blood pressure effects of selective intrarenal versus extrarenal exchange of single chromosome regions between the spontaneously hypertensive rat (SHR) and the normotensive Brown Norway (BN) rat. RESULTS: The replacement of a defined segment of chromosome 1 in the SHR with the corresponding chromosome region of the BN rat was sufficient to attenuate hypertension when selectively achieved either inside the kidney or outside the kidney. CONCLUSIONS: The current finding (1) demonstrates that naturally occurring genetic variants exist that can regulate blood pressure when selectively expressed outside the kidney as well as inside the kidney, and (2) compels reconsideration of the long-held view that in essential hypertension, the genetic transmission of increased blood pressure is determined solely by the genotype of the kidney. PMID- 11473654 TI - Vasopeptidase inhibition attenuates the progression of renal injury in subtotal nephrectomized rats. AB - BACKGROUND: Vasopeptidase inhibitors are a new class of cardiovascular compounds that inhibit both angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP). The aim of the present study was to explore the effects of omapatrilat, a vasopeptidase inhibitor, on renal function and pathology in subtotally nephrectomized (STNx) rats. METHODS: STNx rats were randomized to four groups and treated for 12 weeks: no treatment (N = 14); omapatrilat at a low dose of 10 mg/kg (L, N = 12) and at a high dose of 40 mg/kg (H, N = 10); or an ACE inhibitor, fosinopril, at a dose of 10 mg/kg (N = 12). Sham-operated rats were used as control animals (N = 12). RESULTS: Elevated blood pressure in STNx rats (174 +/- 9 mm Hg) was reduced by omapatrilat in a dose-dependent manner (L, 121 +/- 3 mm Hg; H, 110 +/- 3 mm Hg) and by fosinopril (149 +/- 5 mm Hg). Proteinuria in STNx rats (246 +/- 73 mg/day) was reduced by treatment with fosinopril (88 +/- 21 mg/day) and was normalized by treatment with omapatrilat (L, 30 +/- 4 mg/day; H, 20 +/- 2 mg/day vs. control 25 +/- 1 mg/day). Decreased glomerular filtration rates, elevated plasma urea and creatinine and glomerulosclerosis, and tubulointerstitial fibrosis were ameliorated by omapatrilat and fosinopril to a similar degree. Compared with fosinopril, omapatrilat treatment was associated with increased plasma renin activity and decreased renal ACE and NEP binding in a dose-dependent manner. CONCLUSION: These findings suggest that vasopeptidase inhibition may provide a useful strategy for the treatment of progressive renal disease. PMID- 11473655 TI - Hyperplastic vascular smooth muscle cells of the intrarenal arteries in angiotensin II type 1a receptor null mutant mice. AB - BACKGROUND: Angiotensin II (Ang II), which contracts vascular smooth muscle cells (VSMCs), has been reported to regulate VSMC growth. Recently formed transgenic mice without angiotensinogen or Ang II receptors showed vascular alterations. However, it is still unclear how their VSMCs alter. We explored the role of Ang II via the Ang II type 1a receptor (AT1a) in VSMCs in vivo using AT1a null mutant mice. METHODS: We analyzed the ultrastructure of the intrarenal arteries in AT1a null mutant mice that were homozygous for a targeted disruption of AT1a receptor gene using light and electron microscopy. RESULTS: The structural changes of the intrarenal arteries in AT1a null mutant mice showed the wall thickening, which in the interlobar, arcuate, and proximal interlobular arteries consisted of two additional populations of VSMCs, on the luminal and abluminal sides of the media. The luminal overpopulation of smooth muscle cells (SMCs) was arranged in a longitudinal direction separated by increased interposed elastic laminae. The abluminal overpopulation of SMCs ran in circumferential directions separated from the main population. The cytological structure of VSMCs in AT1a null mutant mice was smaller in size, contained more organelles for protein synthesis and secretion than in control mice, and had poorly developed contractile apparatus. CONCLUSIONS: The lack of AT1a signaling causes structural abnormalities in the renal vascular system and transforms the phenotype of VSMCs into cell proliferation, induces the escape of VSMCs from the circular mechanical integrity, and results in increased synthesis of extracellular matrices. PMID- 11473656 TI - ACE gene polymorphism and IgA nephropathy: an ethnically homogeneous study and a meta-analysis. AB - BACKGROUND: Conflicting results have implicated the angiotensin-converting enzyme (ACE) D allele in the progression of renal damage in patients with IgA nephropathy (IgAN). Most of these findings have been obtained by heterogeneous studies. METHODS: We investigated the ACE insertion/deletion (I/D) gene polymorphism by polymerase chain reaction (PCR) amplification of genomic DNA in an ethnically homogeneous sample size of IgAN patients from Southern Italy. The association between ACE I/D gene polymorphism and the development of the disease was examined in 247 biopsy-proven IgAN patients and 205 healthy subjects. The association with the progression of renal damage was evaluated in 136 patients with a follow-up of > or =3 years according to the slope of the creatinine clearance against time, and in 221 patients with a follow-up of > or =1 year assessing by univariate and multivariate analyses of renal survival. These associations were further estimated in a meta-analysis of seven studies retrieved in the Medline database. The meta-analysis was performed according to the Mantel Haenszel-Peto method when homogeneity of the studies was established using the chi(2) test by Breslow-Day. RESULTS: No difference in the ACE I/D gene distribution between patients and controls and between patients with stable and those with deteriorating renal function was found in our study. A meta-analysis performed separately for Caucasian and Asian studies showed that the ACE I/D gene polymorphism did not contribute to the genetic susceptibility of the development of IgAN (total OR 0.93, 95% CI, 0.71 to 1.23; and 0.95, 95% CI, 0.64 to 1.42, respectively) or the progression of the renal damage (total OR 1.12, 95% CI, 0.67 to 1.88; and 2.26, 95% CI, 0.75 to 6.79, respectively) in both groups. CONCLUSIONS: Our study and meta-analysis suggest caution in the interpretation of results from association studies enrolling heterogeneous populations. Further studies using new tests, which are free of the bias due to population stratification and ethnicity, are warranted. PMID- 11473657 TI - Novel erythropoiesis stimulating protein for treatment of anemia in chronic renal insufficiency. AB - BACKGROUND: Novel erythropoiesis stimulating protein (NESP) is a glycoprotein with a threefold longer terminal half-life than recombinant human erythropoietin (rHuEPO) in humans. The aim of this study was to determine whether NESP is effective for the treatment of anemia at a reduced dosing frequency relative to rHuEPO in patients with chronic renal failure not yet on dialysis [chronic renal insufficiency (CRI)]. METHODS: This was a multicenter, randomized, open-label study. A total of 166 rHuEPO-naive patients with CRI were randomized in a 3:1 ratio to receive NESP (0.45 microg/kg once weekly) or rHuEPO (50 U/kg twice weekly) administered subcutaneously for up to 24 weeks. Dose adjustments were made as necessary to achieve a hemoglobin response, defined as an increase > or =1.0 g/dL from baseline and a concentration > or = 11.0 g/dL. RESULTS: During the 24-week treatment period, 93% (95% CI, 87 to 97%) of patients receiving NESP and 92% (95% CI, 78 to 98%) of patients receiving rHuEPO achieved a hemoglobin response. The median time to response was seven weeks (range of 3 to 25 weeks) in both groups. After correction of anemia, mean hemoglobin concentrations were maintained within the target range of 11.0 to 13.0 g/dL for the remainder of the 24-week treatment period. The safety profiles of NESP and rHuEPO were similar, and no antibodies were detected to either drug. CONCLUSIONS: These results demonstrate that NESP safely and effectively corrects and maintains hemoglobin concentrations at a reduced dosing frequency relative to rHuEPO in patients with CRI, providing a potential benefit to patients and health care providers. PMID- 11473658 TI - Level of hydration and renal function in healthy humans. AB - BACKGROUND: High hydration is commonly used in renal studies to improve the completeness of urine collection. The renal effects of hydration are not well defined. METHODS: Renal function was studied under fasting conditions (baseline) and after a meat meal (2 g of protein/kg body weight) in 12 healthy adults on a low and high hydration regimen of 0.5 and 4 mL of oral water per kg body weight/30 min, respectively. RESULTS: Urine flow, urinary and plasma Na, K, urea, and osmolality were stably different on low and high hydration regimens. At baseline, there were significant or borderline significant correlations of plasma and urine osmolality with glomerular filtration rate (GFR; inulin clearance) only in the low hydration regimen. GFR was higher in the low than the high hydration regimen at all time points. The difference was significant at baseline (19.2%) and at 90 to 180 minutes after the meal (14.4%). After the meal, GFR increased significantly over baseline values only in the high hydration regimen (30.0% at peak time). Urinary excretion of Na, urea, and osmoles was lower in the low than the high hydration regimen at all time points: The difference was significant for Na (at baseline) and osmoles (all time points). Urinary K excretion was not different in the two regimens. After the meal, there were significant increases in urinary excretion of Na (in the low hydration regimen) and urea (90 to 180 min after the meal). CONCLUSIONS: In fasting adults, high hydration lowered GFR and increased natriuresis. After a meat meal, GFR increased only in the high hydration regimen and natriuresis only in the low hydration regimen. Hydration affects GFR and natriuresis under fasting conditions and after a meat meal. PMID- 11473659 TI - Biochemical profile of idiopathic uric acid nephrolithiasis. AB - BACKGROUND: The objective of this study was to elucidate a biochemical profile of patients with idiopathic uric acid nephrolithiasis, without secondary causes (such as dehydration or diarrhea). Study subjects comprised 56 patients with idiopathic uric acid nephrolithiasis (UA stone group) who underwent a full outpatient evaluation. The control group was composed of 54 with absorptive hypercalciuria and 2 normal subjects, matched with the UA stone group according to age, body mass index, and gender. METHODS: Urinary pH and ammonium and serum and urinary uric acid were measured. The fractional excretion of urate was calculated. RESULTS: Compared with the control group, the UA stone group had a significantly higher serum uric acid and significantly lower urinary uric acid, pH (5.34 +/- 0.23 vs. 6.17 +/- 0.36, P < 0.001), and fractional excretion of urate (0.052 +/- 0.028 vs. 0.080 +/- 0.029, P < 0.001), but individual values overlapped considerably between the two groups. Discriminant analysis of the relationship between urinary pH and fractional excretion of urate yielded a "discriminant score," which provided a much better separation between the two groups, with a correct classification in 95.5% of subjects. In contrast, urinary ammonium, citrate, sulfate, and potassium did not differ between two groups. CONCLUSIONS: In idiopathic uric acid nephrolithiasis, urinary pH and fractional excretion of urate are significantly lower than in control subjects, suggestive of defects in urinary acidification and urate excretion. Since these impairments are believed to be associated with primary gout, the underlying disturbance in idiopathic uric acid nephrolithiasis may be primary gout. PMID- 11473660 TI - Clinical performance characteristics of hemodialysis graft monitoring. AB - BACKGROUND: Regular monitoring of dialysis grafts is recommended, but the value of dialysis graft blood flow monitoring and venous pressures in predicting subsequent outcomes are controversial. METHODS: Over a period of one month, we performed simultaneous flow and venous pressure monitoring in 71 dialysis patients with polytetrafluoroethylene (PTFE) grafts. These patients were prospectively followed for one year. Receiver operating characteristic (ROC) curves were constructed to evaluate the performance of the various monitoring techniques. RESULTS: During the period of follow-up, there were 71 graft failures (30 angioplasty alone and 41 thrombosis followed by interventional or surgical revisions). Failed grafts had a lower blood flow rate [799 +/- 452 (SD) mL/min] when compared with those without failure (1019 +/- 485 mL/min, P = 0.05) Single static or dynamic venous-pressure monitoring were not predictive of graft failure. ROC analysis showed poor performance of graft flows in predicting graft failures over the short (30 days, AUC = 0.726, 95% CI, 0.509 to 0.942) and long term (one year, AUC = 0.630, 95% CI, 0.499 to 0.761). An adjustment of graft flows for systolic blood pressure or classification of graft based both on flows and venous pressure did not improve test performance. CONCLUSIONS: Although dialysis graft blood flow rates are statistically different in patients who have graft failure (graft angioplasty and surgery or thrombosis) versus those who do not, the performance characteristics preclude clinical decision-making from an isolated blood flow or venous pressure study. PMID- 11473661 TI - Effect of fluid and sodium removal on mortality in peritoneal dialysis patients. AB - Effect of fluid and sodium removal on mortality in peritoneal dialysis patients. BACKGROUND: Adequacy of peritoneal dialysis (PD) traditionally is assessed using Kt/V(urea) and total creatinine clearance (TCC). However, this approach underestimates the importance of fluid and sodium removal. The aim of this study was to determine the effect of fluid and sodium removal on morbidity and mortality in PD patients. METHODS: One hundred twenty-five PD patients were monitored for three years from the beginning of the treatment. The effects of demographic features, comorbidity, peritonitis rate, blood pressure, medications, blood biochemistry, peritoneal membrane transport characteristics, residual renal function (RRF), Kt/V(urea), TCC, normalized protein nitrogen appearance (nPNA), and removal of sodium and fluid on mortality were evaluated. Total and cardiovascular hospitalization rates were also recorded. A Cox proportional hazards model was used to determine factors predicting mortality. RESULTS: In the Cox model, comorbidity, total sodium and fluid removals, hypertensive status, serum creatinine, and RRF were independent factors affecting survival. In contrast, Kt/V(urea) or TCC did not affect the adjusted survivals. Total sodium and fluid removal and hypertensive status also significantly influenced the hospitalization rate. Systolic and diastolic blood pressures were negatively correlated with total fluid (P < 0.001) and sodium removal (P < 0.001). CONCLUSIONS: Together, these findings suggest that removal of sodium and fluid is a predictor of mortality in PD patients, whereas Kt/V(urea) and TCC are not factors. Adequate fluid and sodium balance is crucial for the management of patients on PD. PMID- 11473662 TI - Sustained low-efficiency dialysis for critically ill patients requiring renal replacement therapy. AB - BACKGROUND: The replacement of renal function for critically ill patients is procedurally complex and expensive, and none of the available techniques have proven superiority in terms of benefit to patient mortality. In hemodynamically unstable or severely catabolic patients, however, the continuous therapies have practical and theoretical advantages when compared with conventional intermittent hemodialysis (IHD). METHODS: We present a single center experience accumulated over 18 months since July 1998 with a hybrid technique named sustained low efficiency dialysis (SLED), in which standard IHD equipment was used with reduced dialysate and blood flow rates. Twelve-hour treatments were performed nocturnally, allowing unrestricted access to the patient for daytime procedures and tests. RESULTS: One hundred forty-five SLED treatments were performed in 37 critically ill patients in whom IHD had failed or been withheld. The overall mean SLED treatment duration was 10.4 hours because 51 SLED treatments were prematurely discontinued. Of these discontinuations, 11 were for intractable hypotension, and the majority of the remainder was for extracorporeal blood circuit clotting. Hemodynamic stability was maintained during most SLED treatments, allowing the achievement of prescribed ultrafiltration goals in most cases with an overall mean shortfall of only 240 mL per treatment. Direct dialysis quantification in nine patients showed a mean delivered double-pool Kt/V of 1.36 per (completed) treatment. Mean phosphate removal was 1.5 g per treatment. Mild hypophosphatemia and/or hypokalemia requiring supplementation were observed in 25 treatments. Observed hospital mortality was 62.2%, which was not significantly different from the expected mortality as determined from the APACHE II illness severity scoring system. CONCLUSIONS: SLED is a viable alternative to traditional continuous renal replacement therapies for critically ill patients in whom IHD has failed or been withheld, although prospective studies directly comparing two modalities are required to define the exact role for SLED in this setting. PMID- 11473663 TI - Endothelial cell protection against ischemia/reperfusion injury by lecithinized superoxide dismutase. AB - BACKGROUND: Organs used for transplantation may experience long periods of cold ischemic preservation and consequently oxygen free radical-mediated damage following reperfusion. Lecithinized superoxide dismutase (lec-SOD) is a novel free radical scavenger that has been shown to bind with high affinity to cell membranes. The aim of this study was to determine whether lec-SOD bound to endothelial cells under organ preservation conditions to mediate direct antioxidant activity at the endothelial cell surface and thus offer protection against the harmful effects of ischemia/reperfusion injury. METHODS: An in vitro study was performed on large vessel endothelial cells (HUVEC) and a human microvascular endothelial cell line HMEC-1, to investigate the potential therapeutic benefits of incorporating lec-SOD into organ preservation solution. A cold hypoxia/reoxygenation system was developed to examine lec-SOD binding affinity to endothelial cells, protection against hypoxia/reoxygenation-induced cell death, and neutrophil adhesion. RESULTS: Lec-SOD bound to endothelial cells with higher affinity than unmodified recombinant human superoxide dismutase (rhSOD) and significantly protected both HUVEC and HMEC-1 from cell death following 27 hours of cold hypoxia (P < 0.01). Furthermore, neutrophil adhesion to the endothelium stimulated by hypoxia and reoxygenation was significantly inhibited by treatment with lec-SOD but not by lecithin or rhSOD (P < 0.01). Analysis by flow cytometry demonstrated that E-selectin and ICAM-1 were up regulated by hypoxia/reoxygenation that was inhibited in part by lec-SOD. CONCLUSIONS: The results from this study suggest that incorporation of lec-SOD into organ preservation solutions provides effective protection to endothelial cells against cold ischemia and reperfusion injury following transplantation. PMID- 11473664 TI - Beyond a glomerulocentric view of inflammation. PMID- 11473665 TI - Thin glomerular basement membrane disease. PMID- 11473666 TI - Need for publication of all studies testing the SphygmoCor device. PMID- 11473668 TI - Elevated blood pressure predicts risk of acute rejection in renal allograft recipients. PMID- 11473670 TI - Preventing dialysis hypotension: a comparison of usual protective maneuvers. PMID- 11473672 TI - Drug-induced acute interstitial nephritis. PMID- 11473676 TI - Definition of respiratory symptoms and disease in early childhood in large prospective birth cohort studies that predict the development of asthma. AB - We have reviewed the prospective value of early respiratory symptoms for determining the risk of development of asthma later in life by using data from studies based on the general population, hospital population, and general practices. Although "wheezing" in infancy generally has a good prognosis, it is an important risk factor for the development of asthma later in life. The prognostic value of "coughing" and "shortness of breath" in infancy for the later development of asthma is less clear. Despite the fact that no internationally accepted criteria for the definition of asthma in early childhood are available, many studies have been performed on this topic. We also investigated the outcome variables that were used to describe respiratory symptoms and disease in early childhood in the publications of nine large prospective birth cohort studies on the development of asthma. From seven of these studies, we reviewed the original questionnaires. We found that various studies used different outcome variables, but the data actually collected were similar. This is an important observation because it implies that comparisons between studies can be markedly improved by data sharing among investigators. PMID- 11473677 TI - Asthma severity and inflammation markers in children. AB - The relationship of airway inflammation with asthma severity remains unclear. Our aim was to correlate the results of recommended methods of assessment of inflammation with measures of asthma control, in children with a wide range of asthma severity. The study was a cross-sectional investigation of 58 children receiving a wide range of treatment, including 10 treated without regular maintenance therapy and 29 treated with high-dose inhaled corticosteroids (CS). Exhaled nitric oxide (NO), serum eosinophil cationic protein (ECP), and induced sputum (processed for eosinophil count and ECP level) were related to recent symptoms, lung function, and bronchial responsiveness. There was no significant correlation between the results of any METHOD: Neither did any marker of airway inflammation relate to recent symptoms, unlike PC20, which did. There was a significant, inverse correlation between the forced expiratory volume in 1 s (FEV1) and both NO and sputum ECP (r=-0.46, p=<0.001; r=-0.48, p=0.004, respectively). Sputum eosinophils were inversely related to the dose of methacholine that corresponded to a 20% fall in FEV1 (PC20) (r=-0.57, p=0.02). Serum ECP did not relate to any measure of asthma control. There was no association of any recommended inflammation markers with current symptoms and only a weak relationship between them and physiological measures. The place of these markers remains unclear and their use in clinical practice needs further investigation by long-term longitudinal studies. PMID- 11473678 TI - Development of specific immunoglobulin E in coughing toddlers: a medical records review of symptoms in general practice. AB - The aim of this study was to study whether young children, originally immunoglobulin E (IgE) negative and who became sensitized to specific inhalation allergens, presented more frequently to their general practitioner (GP) with other allergy- and asthma-related symptoms than children who remained IgE negative. It was also investigated whether asthma was diagnosed more often in children who developed IgE to inhalant allergens. Coughing children, 1-5 years of age, visiting the participating GPs, were tested for IgE antibodies to mites, dogs, and cats by using radioallergosorbent testing (RAST). All IgE-negative (RAST < 0.2 IU/ml) children were re-tested after 2 years. The medical records of 162 children were reviewed on asthma- and allergy-related symptoms and on prescribed medication. After 30 months, 27 of the 162 children (17%) had become IgE positive for one or more allergens. Most children (93%) had visited their GP for treatment of respiratory symptoms during this period. However, the children who had become IgE positive had visited their GP more often than the children who remained IgE negative. Differences in visits were seen for: shortness of breath (52% IgE-positive vs. 19% IgE-negative children, respectively), wheeze (37% vs. 17%), allergic rhinitis (33% vs. 16%), and pneumonia (22% vs. 8%), but not for coughing (89% vs. 88%). The IgE-positive children were more frequently diagnosed by their GP as having asthma (48%) than were the IgE-negative children (23%). In a multivariate analysis, indicators of becoming IgE positive were: a visit for shortness of breath (odds ratio [OR] = 6.9; 95% confidence interval [CI] = 2.1 23.1) and two or more visits for wheeze (OR = 6.0; 95% CI = 1.9-19.2), adjusted for breast-feeding, age, and asthma or allergy in the family. The positive predictive value (PPV) of being IgE positive with a diagnosis of asthma was 90% (whereas the negative predictive value was 48.0%) for a child attending their GP for treatment of wheeze. For recurrent coughing (six or more visits) and shortness of breath, the PPVs were 73% and 71%, respectively. The development of sensitization to common inhalant allergens is associated with specific allergy and asthma-related symptoms in young children. IgE-positive children were more frequently diagnosed as having asthma by their GP. This implies that in general practice it is possible to detect children at high risk for developing allergic asthma early in life by their respiratory symptoms and by subsequent testing for specific IgE to inhalant allergens. PMID- 11473679 TI - Developmental follow-up in 15-month-old infants of asthmatic vs. control mothers. AB - The purpose of this study was to evaluate mental and psychomotor development in infants of mothers whose asthma was actively managed during pregnancy and to compare the results with those froms infants of non-asthmatic mothers. Bayley Scales were assessed at age 15 +/- 3 months in 379 infants of asthmatic mothers and 376 control infants. Relationships were assessed between developmental indices and asthma severity, socioeconomic status, and infant prematurity. No significant differences in developmental indices were observed between infants of asthmatic mothers and control infants. No relationships were identified between developmental indices and maternal asthma severity. In the infants of both asthmatic and control mothers, a lower mean psychomotor developmental index was associated with birth weight < 2,500 g, and a lower mental developmental index with lower socioeconomic status. Hence, infants of asthmatic mothers whose asthma has been actively managed during pregnancy have developmental outcomes at 15 months of age that are similar to those of control infants. PMID- 11473680 TI - Montelukast in the treatment of children with moderate-to-severe atopic dermatitis: a pilot study. AB - The primary action of leukotrienes includes contraction of human airway muscle, chemotaxis, and increased vascular permeability, with secondary effects of inhibiting allergen-induced early and late responses. Although there is limited available information and research regarding leukotrienes in atopic dermatitis (AD), there is evidence to support their role in the pathogenesis of the disease. We conducted a pilot study to test the efficacy of montelukast, a cysteinyl leukotriene-1 receptor antagonist, in 15 patients (6-16 years of age) with moderate-to-severe AD, using a randomized double-blind placebo-controlled crossover study. These patients had chronic moderate-to-severe AD, despite being on conventional therapy. They were randomized either to placebo for 4 weeks and then the study drug for 4 weeks, or vice versa. There was a 2-week run-in period for all participants before commencement of the study, and a 2-week washout before crossover. At enrollment and on each follow-up visit, every patient was assessed by a single observer and objectively scored for disease extent and severity. A subjective score was given for the impact of eczema on daily living. There was statistical improvement in patents on active treatment compared with placebo in the severity of AD (p < 0.05). Our findings suggest that leukotriene receptor antagonist as an adjunct treatment has an anti-inflammatory effect on moderate-to-severe AD. A larger trial is needed to ascertain its efficacy fully. PMID- 11473681 TI - Nebulized budesonide after hospitalization for recurrent bronchial obstruction in children younger than 18 months. AB - A multi-center, double-blind, randomized dose-response study was performed to assess the effect of 3 months of treatment with two different doses of inhaled nebulized budesonide in children with acute recurrent bronchial obstruction (BO) causing hospitalization. Steroid-naive children younger than 18 months were included when admitted to hospital because of BO for at least the second time, and were followed-up monthly for 15 months. Forty-five of 49 subjects (43 boys, 2 girls) (mean age 9.3 months upon inclusion) completed the study. Twenty-four patients (20 boys, 4 girls) received nebulized budesonide 0.5 mg twice daily for 1 month followed by 0.25 mg daily for the next 2 months, whereas 25 children received 0.1 mg twice daily throughout the 3-month treatment period. Outcome (number of BO episodes, time to first BO after start of treatment, and use of rescue medication), as well as height/length and weight, were assessed at the start of treatment and monthly for the following 3 months, as well as for 12 months after cessation of treatment (15 months in total). There was an overall tendency towards better symptom control (fewer episodes of acute BO during treatment and follow-up, fewer hospital visits because of acute BO, lower clinical score during follow-up, and less use of rescue medication during follow up) in the high-dose treatment group vs. the low-dose treatment group. However, the differences did not reach statistical significance for any of the outcomes. The only significant difference in effect between the groups was fewer children in the high-dose group treated openly with nebulized budesonide during follow-up. Length/height and weight gain did not differ significantly between the two treatment groups throughout the study. There was no significant dose-dependent beneficial effect of 3 months of treatment with nebulized budesonide in infants and toddlers with at least two hospitalizations for acute bronchial obstruction. PMID- 11473682 TI - Complement analysis in children with idiopathic membranoproliferative glomerulonephritis: a long-term follow-up. AB - Fifty children with idiopathic membranoproliferative glomerulonephritis (MPGN), aged 2-14 years at apparent onset, were monitored for the presence of C3 nephritic factor (C3 NeF) and signs of complement activation in serum. In addition, C3 allotyping was performed in 32 patients. Observation time ranged from 2 to 20 (median 11) years. C3 NeF activity was detected at least once in 60% of the patients (in 11 of 26 with type I, in 15 of 17 with type II, and in four of seven with type III). C3 NeF-positive patients had significantly reduced levels of CH50 and C3 and elevated levels of C3dg/C3d. During follow-up, C3 levels were persistently normal in 62% of the patients with MPGN type I and in 43% with type III but in only 18% with type II. C3 allotype frequencies differed from those found in healthy controls with a significant shift to the C3F/C3FS variants in C3 NeF-positive patients. C3b(Bb)P as a marker for alternative pathway activation was not increased in C3 NeF-positive patients. Despite the presence of C3 NeF activity, C3 levels remained normal in six patients throughout the observation period. C3 NeF became undetectable in six patients, whereas seven developed C3 NeF activity during follow-up. There was no significant difference in renal survival probability in patients with or without C3 NeF activity. Neither C3 variants nor continuous low C3 or low CH50 levels had any prognostic value for the clinical outcome. No factor H deficiency was detected. PMID- 11473683 TI - Re.: Findings in the retrospective study of hospital admissions for asthma in Finland. PMID- 11473685 TI - Oil bodies and their associated proteins, oleosin and caleosin. AB - Oil bodies are lipid storage organelles which have been analyzed biochemically due to the economic importance of oil seeds. Although oil bodies are structurally simple, the mechanisms involved in their formation and degradation remain controversial. At present, only two proteins associated with oil bodies have been described, oleosin and caleosin. Oleosin is thought to be important for oil body stabilization in the cytosol, although neither the structure nor the function of oleosin has been fully elucidated. Even less is known about caleosin, which has only recently been described [Chen et al. (1999) Plant Cell Physiol 40: 1079 1086; Naested et al. (2000) Plant Mol Biol 44: 463-476]. Caleosin and caleosin like proteins are not unique to oil bodies and are associated with an endoplasmatic reticulum subdomain in some cell types. Here we review the synthesis and degradation of oil bodies as they relate to structural and functional aspects of oleosin and caleosin. PMID- 11473686 TI - Small complex-type N-linked glycans are attached to cell-wall bound exo-beta glucanases of both mung bean and barley seedlings. AB - N-linked glycans of wall-bound exo-beta-glucanases from mung bean and barley seedlings, namely Mung-ExoI and Barley-ExoII, were characterized. The N-linked glycans of Mung-ExoI and Barley-ExoII were liberated by gas-phase hydrazinolysis followed by re-N-acetylation. Their structures were determined by two-dimensional sugar-mapping analysis and MALDI-TOF mass spectrometry. N-glycans from both glucanases were of paucimannosidic-type (small complex-type) structures, Manalpha1-6(+/-Manalpha1-3)(Xylbeta1-2)Manbeta1-4GlcNAcbeta1-4(+/-Fucalpha1-3) GlcNAc, which are known as typical vacuole-type N-glycans. The results suggest that N-glycans of cell-wall glucanase were produced by partial trimming of complex-type N-glycans by exoglycosidases during its transport from Golgi apparatus to cell walls or in the cell walls. PMID- 11473687 TI - Stimulation of plant plasma membrane Ca2+-ATPase activity by acidic phospholipids. AB - The effect of phospholipids on the activity of the plasma membrane (PM) Ca2+ ATPase was evaluated in PM isolated from germinating radish (Raphanus sativus L. cv. Tondo Rosso Quarantino) seeds after removal of endogenous calmodulin (CaM) by washing the PM vesicles with EDTA. Acidic phospholipids stimulated the basal Ca2+ ATPase activity in the following order of efficiency: phosphatidylinositol 4,5 diphosphate (PIP2) approximately phosphatidylinositol 4 monophosphate>phosphatidylinositol approximately phosphatidylserine approximately phosphatidic acid. Neutral phospholipids as phosphatidylcholine and phosphatidylethanolamine were essentially ineffective. When the assays were performed in the presence of optimal free Ca2+ concentrations (10 &mgr;M) acidic phospholipids did not affect the Ca2+-ATPase activated by CaM or by a controlled trypsin treatment of the PM, which cleaved the CaM-binding domain of the enzyme. Analysis of the dependence of Ca2+-ATPase activity on free Ca2+ concentration showed that acidic phospholipids increased Vmax and lowered the apparent Km for free Ca2+ below the value measured upon tryptic cleavage of the CaM-binding domain; in particular, PIP2 was shown to lower the apparent Km for free Ca2+ of the Ca2+-ATPase also in trypsin-treated PM. These results indicate that acidic phospholipids activate the plant PM Ca2+-ATPase through a mechanism only partially overlapping that of CaM, and thus involving a phospholipid-binding site in the Ca2+-ATPase distinct from the CaM-binding domain. The physiological implications of these results are discussed. PMID- 11473688 TI - Purification and characterization of an aspartic protease from potato leaves. AB - A protease was isolated from potato (Solanum tuberosum L. cv. Pampeana) leaves 48 h after detaching, when aspartic protease (AP) activity is markedly increased. Purification was performed by ammonium sulfate precipitation, ion exchange chromatography and affinity chromatography. A size of 40 kDa was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; it is monomeric and its properties are consistent with those of aspartic proteinases (EC 3.4.23): it has a pH optimum of 3 and it is inhibited by pepstatin. Like other plant APs, leaf AP appears to be glycosylated with a complex-type N-glycan. The enzyme has properties different from those of a tuber AP previously described, indicating that they may have different physiological roles. PMID- 11473689 TI - Mitochondrial alternative oxidase acts to dampen the generation of active oxygen species during a period of rapid respiration induced to support a high rate of nutrient uptake. AB - When wild type (wt) tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) suspension cells were grown under phosphate (P) limitation, they contained large amounts of mitochondrial alternative oxidase (AOX). When these cells were resupplied with P, there was a large, immediate and sustained stimulation of respiration to support a period of rapid P uptake. Two lines of evidence suggest that the abundant level of AOX present in wt cells contributed to this stimulated rate of respiration. First, when P-limited transgenic antisense tobacco cells (AS8) lacking AOX were resupplied with P, the stimulation of respiration was much less dramatic even though these cells displayed similar rates of P uptake. Second, while the stimulated rate of respiration in AS8 cells was insensitive (as expected) to the AOX inhibitor n-propyl gallate (nPG), much of the stimulated rate of respiration in wt cells could be inhibited by nPG. Given the non phosphorylating nature of AOX respiration, wt cells required higher rates of electron transport to O2 than AS8 cells to support similar rates of P uptake. The utilization of AOX by wt cells during P uptake was apparently not occurring because the cytochrome (Cyt) pathway alone could not fully support the rate of P uptake, as the respiration of cells lacking AOX (either untreated AS8 cells or wt cells treated with nPG) supported similar rates of P uptake as wt cells with abundant AOX. Rather, we provide in vivo evidence that the utilization of AOX during the period of high respiration supporting P uptake was to dampen the mitochondrial generation of active oxygen species (AOS). PMID- 11473690 TI - Effect of embryonic axis removal and exogenous calcium on carboxypeptidase I of mung bean seedling cotyledons. AB - Removal of the embryonic axis prevents the normal decline of carboxypeptidase (Cpase) I in mung bean seedling cotyledons. Cpase I activity and protein, the latter manifested on western blots, almost completely disappear about 24 h before the cotyledon abscises. Of the 3 proteolytic enzyme patterns, only that of Cpase I can be restored by an exogenous supply of 10 mM CaCl2 in the agar growth medium. The calcium effect is dependent on [CaCl2] and is not manifested in the presence of chelators and calcium channel blockers. For detached cotyledons to show the normal low level of Cpase I by the eighth day of growth, calcium had to be supplied during seed imbibition and throughout the entire time from removal of the axis. The difference between detached cotyledons in the absence and presence of calcium was greatest when the cotyledons were detached 4-6 days after seed imbibition. Loss of Cpase I activity and protein can be demonstrated in vitro, with the maximum level of Cpase I-degrading activity measured 4 days after seed imbibition under the same growth conditions used to study the calcium effect. It is sensitive to pepstatin and has a pH optimum of 3, suggesting that this Cpase I degrading activity is due to an aspartic protease. PMID- 11473691 TI - Phosphoenolpyruvate carboxylase in mistletoe leaves: Regulation of gene expression, protein content, and covalent modification. AB - Seasonal changes in the activity of phosphoenolpyruvate carboxylase (PEPCase, EC 4.1.1.31), a key enzyme in the interaction of carbohydrate and nitrogen metabolism, were studied in leaves of the C3 semiparasitic mistletoe, Viscum album, growing on different host trees. Maximum extractable PEPCase activities were higher in leaves of mistletoes growing on Betula pendula and Alnus glutinosa hosts compared with those on the conifers, Abies alba and Larix decidua. Independent of host, maximum extractable PEPCase activities were high in spring and autumn while low in summer. Samples with higher PEPCase activities showed higher amounts of PEPCase protein and higher PEPCase mRNA levels. A curvilinear correlation between leaf total nitrogen content and the maximum extractable PEPCase activity as well as PEPCase mRNA level suggested that nitrogen might affect the activity of PEPCase of mistletoe by up-regulating gene expression. In addition to extractable activity, seasonal changes of the PEPCase activation state, the ratio of activities resulting from limited:non-limited assays, were found, which was correlated to the variation of malate content in leaves of mistletoe. ATP-dependent activation of PEPCase was characterized by an increase in I0.5(L-malate), indicating that PEPCase of leaves of mistletoes is probably regulated via phosphorylation. PMID- 11473692 TI - Changes in cell-wall properties of wheat (Triticum aestivum) roots during aluminum-induced growth inhibition. AB - The effects of aluminum (Al) on root elongation, the mechanical extensibility of the cell wall, and the amount of cell-wall polysaccharides in the roots of Al resistant (Atlas 66) and Al-sensitive (Scout 66) cultivars of wheat (Triticum aestivum L.) were examined. Exposure to 10 &mgr;M AlCl3 for 6 h inhibited root elongation in Scout 66 but not in Atlas 66. It also decreased the mechanical extensibility of the cell wall in the roots of both cultivars, but prominently only in the roots of Scout 66. The amount of hemicellulose in the 10-mm region of root apex of Scout 66 was increased by the exposure to Al, especially in the apical regions. Al did not influence the neutral sugar composition of either pectin or hemicellulose in Scout 66 roots. However, Al increased the weight average molecular mass of hemicellulosic polysaccharides and the amounts of wall bound ferulic and diferulic acids in Scout 66 roots. These findings suggest that Al modifies the metabolism of cell-wall components and thus makes the cell wall thick and rigid, thereby inhibiting the growth of wheat roots. PMID- 11473693 TI - Genes encoding chitinase-antifreeze proteins are regulated by cold and expressed by all cell types in winter rye shoots. AB - One group of antifreeze proteins (AFPs) is composed of two chitinases that accumulate in the apoplast of winter rye leaves during cold acclimation. In this study, the 28- and 35-kDa chitinase-AFPs were localized in nonacclimated and cold acclimated rye leaves by immunoelectron microscopy with an antiserum produced against the purified winter rye 35-kDa chitinase-AFP. In cold-acclimated winter rye leaves, labelled chitinase-AFPs were abundant in the walls of epidermal, parenchymal sheath and mesophyll cells and xylem vessels, while less label was present in walls of vascular parenchyma cells. In contrast, chitinase labelling was essentially absent in the nonacclimated cells except in xylem vessels. As shown by RNA blotting, the transcripts of chitinase-AFPs accumulated to a high level in rye leaves during cold acclimation, to a lesser extent in crowns and were not detectable in roots. mRNA transcripts of the 28-kDa chitinase-AFP were localized in rye leaves by in situ hybridization. The chitinase-AFP transcripts were found in the same cell types as the protein itself. We conclude that all metabolically active cell types in cold-acclimated winter rye leaves and crowns are able to synthesize chitinase-AFPs and secrete them into adjacent cell walls, where they may interact with ice to delay its propagation through the plant and modify its growth. PMID- 11473694 TI - Water sorption-desorption in conifer cuticles: The role of lignin. AB - Current information on the type and amount of biopolymers present in the epidermis of conifer species is still insufficient. This work presents the detailed morphology and chemical composition of Araucaria bidwillii cuticle after selective treatments to remove the different types of biopolymers. After removal of the waxes, cutin and polar hydrolyzable components, a lignin-like fraction, which makes up 25% of the initial cuticle weight, was identified by GC-MS and infrared spectroscopy. The isolated lignin is of G type, mainly formed by guaiacyl units. This composition indicates that the conifer cuticle investigated here has similar composition to other conifer-isolated cuticles. Water sorption and desorption by the isolated cuticle and the different cuticle fractions, including lignin, were studied. The analysis of the isotherms, following distinct physicochemical models, gave useful information on the structural and physiological role of the different biopolymers present in the cuticle. Lignin fraction showed both a high water sorption and capability of retaining it in comparision to other cuticle components. Hysteresis effect on water sorption desorption cycle and water cluster formations has also been studied, and their physiological role discussed. PMID- 11473695 TI - Limited carbohydrate availability as a potential cause of fruit abortion in Rubus chamaemorus. AB - Fruit abortion can be caused by a range of abiotic and biotic factors. To gain a better understanding of the causes of the high fruit abortion frequency in cloudberry (Rubus chamaemorus L.), we manipulated different sources of carbon, that is, leaves and rhizome. We also manipulated flower number to see if competition between floral ramets explained fruit abortion in cloudberry. Reducing the number of flowers had no impact on fruit abortion frequency. In fact, the species forms an extensive rhizome network with only a few ramets per clone and competition between floral ramets is unlikely. Ramet defoliation had limited impact on fruit abortion, but successful fruit development was affected by rhizome length. The longer the rhizome, the higher the chances to mature a fruit. These results suggest that current photoassimilate production by the reproductive ramet alone is insufficient to insure fruit development. Carbon can come from other ramets but distances are usually high between ramets. Fruit production might thus depend on the use of stored carbohydrates in the rhizome to balance insufficient photosynthetic contribution during fruit production. PMID- 11473696 TI - The promoter of a cytosolic glutamine synthetase gene from the conifer Pinus sylvestris is active in cotyledons of germinating seeds and light-regulated in transgenic Arabidopsis thaliana. AB - We have isolated and characterized a genomic clone encoding Scots pine (Pinus sylvestris) cytosolic glutamine synthetase (GS). The clone contains the 5' end half of the gene including part of the coding region and 980 bp upstream of the translation initiation codon. The major transcription start site (+1) was mapped around 180 nucleotides upstream of the translation initiation codon. Sequence analysis of the 5'-upstream region of the gene reveals the presence of putative regulatory elements including a poly-CT consensus sequence, a purine-rich tandem repeat and two AT-rich regions. Fusions of the upstream gene region to uidA were shown to be transiently expressed in the cotyledons of germinating pine seeds transformed by microprojectile bombardment. Stable transformation of Arabidopsis thaliana revealed the shoot apical meristem as the major region of heterologous permanent expression in Arabidopsis, in agreement with the expression of the GS gene in Pinus. Moreover, quantitative data derived from fluorometric beta glucuronidase assays in control and continuous light-grown transgenic Arabidopsis plants indicate that the isolated upstream region of the gene contains regulatory sequences involved in the response to light. PMID- 11473697 TI - Isolation of aluminum-tolerant cell lines of tobacco in a simple calcium medium and their responses to aluminum. AB - Aluminum (Al)-tolerant cell lines (ALT301 and ALT401) of tobacco were isolated in a simple calcium (Ca) solution from ethyl methane sulfonate (EMS)-treated suspension cultured tobacco cells (Nicotiana tabacum L. cv. Samsun, a cell line SL) at the logarithmic phase of growth. A highly tolerant cell line ALT301 exhibited the accumulation of Al and the deposition of callose to the same extent as the parental SL cells during the exposure to Al. However, the Al-treated ALT301 cells grew much better than the Al-treated SL cells after transfer to Al free growth medium. Compared to SL cells, ALT301 cells were more tolerant to toxicity of copper and iron, but not to that of lanthanum. These results suggest that ALT301 cells have an internal tolerance mechanism, which makes cells grow normally in spite of Al accumulation and Al-induced lesion represented by the deposition of callose. This tolerance mechanism seems also to be effective against copper and iron toxicity. A slightly tolerant cell line ALT401 also accumulated Al to the same degree as SL cells, but deposited significantly less callose than did SL cells (43% of SL). The growth of ALT401 cells after Al treatment was only slightly better than that of SL cells. Thus, it seems that ALT401 cells have a mechanism to protect cells only from the Al-induced deposition of callose, which is not enough to overcome the Al-induced inhibition of growth. The different phenotypes exhibited by these Al-tolerant cell lines suggest that the deposition of callose is not directly related to the inhibition of growth in Al-treated cells. PMID- 11473698 TI - Stability of barley aleurone transcripts: Dependence on protein synthesis, influence of the starchy endosperm and destabilization by GA3. AB - We have studied the stability of Barley aleurone and embryo expressed (Balem) transcripts in aleurone layers. The Per1, Ole1 and Ole2 transcripts are abundant during desiccation and in dry resting seeds, while B12D and B22E transcripts are expressed mainly during seed maturation and germination. From 21 to 40 days post anthesis (DPA) incubation of aleurone layers resulted in a substantial, but differential reduction in the levels of these transcripts. In contrast, Balem transcript levels in aleurone layers of incubated embryoless grains were (except for B22E) similar to those of freshly dissected layers. Cycloheximide lowered transcript levels significantly. This indicates that a protein-synthesis dependent mRNA-stabilizing mechanism is active in the aleurone cells when attached to the starchy endosperm. At the onset of seed desiccation (40 DPA), half-lives of transcripts to be stored in the dry seed were up to several days longer than the half-life of B22E, which decreases during seed maturation. While the Per1, Ole1 and Ole2 transcript levels decline rapidly in the aleurone layers of mature, germinating seeds, the genes are actively transcribed and their transcripts highly stable in the aleurone of incubated embryoless seeds. The expression of Ole1 and Ole2, as well as Per1, can be repressed 100-1 000-fold by gibberellic acid (GA3) in a dose-dependent manner. Abscisic acid can counteract the GA3 repression. Incubations with transcriptional and translational inhibitors indicate that GA3 inhibits the transcription of these genes and at the same time induces a protein-synthesis-dependent mechanism destabilizing their mRNA molecules present. PMID- 11473699 TI - The nitrogen-induced recovery of alpha-zein gene expression in in vitro cultured opaque2 maize endosperms depends on the genetic background. AB - The effect of nitrogen nutrition on the accumulation of seed storage proteins has been studied in vitro by cultivating on agar media maize (Zea mays L.) endosperm explants from seeds at 10 days after pollination. The experiments were performed on various genetic backgrounds bearing different opaque2 (o2) mutant alleles and on the corresponding wild-type lines. In the seed of the o2 genotypes the high molecular weight alpha-zein polypeptides (zHs), whose transcription is Opaque2 (O2) regulated, are absent or extremely reduced. The endosperms were incubated on basal agar medium with amino acid supply. In these growth conditions, fresh and dry weights increased in both wild-type and o2 endosperms, irrespective of the genetic background. In 4 out of the 5 o2 mutant genotypes analysed we detected an accumulation of the zHs similar to the corresponding wild-type explants or seeds. However, in one of these mutants, Mo17o2R, the addition of amino acids to the culture media had no effect on the zH accumulation. We showed that the Mo17o2R behaviour is not due to a negative regulation but to the absence of putative transcription factor(s) able to regulate the zH transcription occurring in the other o2 mutants. PMID- 11473700 TI - Effects of cytokinin on ethylene production and nodulation in pea (Pisum sativum) cv. Sparkle. AB - In this study, we were interested in learning if cytokinins play a role in the developmental process that leads to nodulation in the pea cv. Sparkle. We demonstrate that the application of the synthetic cytokinin BAP (6-benzyl-amino purine) results in a number of nodulation-related changes. BAP stimulates the production of ethylene, a known inhibitor of nodulation. At low levels (up to 1 &mgr;M), BAP also stimulates nodulation but as its concentration is increased (up to 25 &mgr;M), nodule number decreases. In BAP-treated roots, the infection threads are abnormal; they are twisted, very knotty, and generally grow in a direction parallel to the root surface. In addition, the centers of cell division in the inner cortex are very few. Thus, BAP-treated Sparkle appears to phenocopy the low-nodulating pea mutant R50 [Guinel FC, Sloetjes LL (2000) Ethylene is involved in the nodulation phenotype of Pisum sativum R50 (sym 16), a pleiotropic mutant that nodulates poorly and has pale green leaves. J Exp Bot 51: 885-894]. However, it appears doubtful that there is a direct correlation between the actions of cytokinin and ethylene in causing a reduction in nodule organogenesis because nodulation is not restored by treating BAP-treated Sparkle with ethylene inhibitors. PMID- 11473701 TI - Gibberellins in relation to flowering in Polianthes tuberosa. AB - Endogenous gibberellins (GAs) in corms of Polianthes tuberosa L. (cv. Double) were isolated and identified by high performance liquid chromatography, bioassay and combined capillary gas chromatography-mass spectrometry (GC-MS). Gibberellins A1, A19, A20 and A53 were quantified at the vegetative, early floral initiation and flower development stages. The identification of 13-hydroxylated GAs indicates the presence of the early 13-hydroxylation pathway in P. tuberosa corms. An increase in GA1 and GA20, and a decrease in GA19 levels, coincided with the transition from the vegetative phase to the stages of early floral initiation and flower development. GA53 stayed at constant levels at the 3 different growth stages. The absence of GA1 in vegetative corms and its presence in corms at early floral initiation and flower development stages suggest that GA1 is a causal factor in inducing floral initiation in P. tuberosa. When GA1, GA3, GA4, GA20 and GA32 were applied to corms at the vegetative stage (plants about 5 cm in height), floral initiation was promoted by all of the GAs used, GA32 being the most active. In contrast with the other GAs, GA32 had no effect on stem elongation. Therefore, it is suggested that hydroxylated C-19 GAs play an important role in flower induction in P. tuberosa. PMID- 11473702 TI - Photocontrol of flowering and stem extension of the intermediate-day plant Echinacea purpurea. AB - Intermediate-day plants (IDP) flower most rapidly and completely under intermediate photoperiods (e.g., 12-14 h of light), but few species have been identified and their flowering responses are not well understood. We identified Echinacea purpurea Moench as an IDP and, based on our results, propose a novel mechanism for flowering of IDP. Two genotypes of E. purpurea ('Bravado' and 'Magnus') flowered most completely (>/=79%) and rapidly and at the youngest physiological age under intermediate photoperiods of 13-15 h. Few (/=230% as the photoperiod or NI duration increased, until plants received a saturating duration (at 14 or 1 h, respectively). Flowering was inhibited when 16-h photoperiods were deficient in red (R, 600-700 nm) light, and was promoted when photoperiods were deficient in far-red (FR, 700-800 nm) light. Because of our results, we propose the flowering behavior of IDP such as E. purpurea is composed of two mechanisms: a light-dependent response operating through light-labile (type I) phytochrome in which flowering is inhibited by an LD, and a light-stable (type II) phytochrome (i.e., phyB, D and E) response in which flowering is promoted by a short-night. PMID- 11473703 TI - Developmental expression and perturbation of arabinogalactan-proteins during seed germination and seedling growth in tomato. AB - Arabinogalactan-proteins (AGPs) are a family of highly glycosylated hydroxyproline-rich glycoproteins present throughout the plant kingdom. A synthetic chemical reagent, (beta-D-Gal)3 Yariv reagent, specifically binds AGPs and can be used for histochemical staining, isolating and probing the function of AGPs. Here, the role of AGPs in tomato (Lycopersicon esculentum Mill. cv. UC82B) seed germination and seedling growth was examined by following expression of AGPs during these events and by treatment with (beta-D-Gal)3 Yariv to perturb AGP function. AGP expression changed during germination and seedling development both quantitatively and qualitatively as revealed by analysis of total AGP content, crossed electrophoresis patterns, RNA blots using LeAGP-1 probe, and western blots with LeAGP-1, JIM13, and MAC207 antibodies. (beta-D-Gal)3 Yariv treatment of seeds and developing seedlings did not affect percent seed germination, but markedly inhibited seedling growth in roots and to a lesser degree in shoots. Root growth inhibition encompassed reductions in overall root length, epidermal root cell elongation, root cell numbers and root hair formation. This growth inhibition was reversible following removal of (beta-D-Gal)3 Yariv. In a related experiment, water uptake by tomato seedlings was greatly inhibited by (beta-D Gal)3 Yariv treatment. Based on these experiments, AGPs are clearly associated with tomato seedling development and likely to function in root growth, more specifically in cell elongation, cell proliferation, root hair formation and water uptake. PMID- 11473704 TI - Ubiquitin- and proteasome-dependent proteolysis in plants. AB - In recent years it has become obvious that protein degradation is an important catabolic process during development in plants and animals. One very conserved degradative system is the ubiquitin- and proteasome-dependent proteolytic pathway, which is found in all eukaryotes from yeast to mammals and plants. The pathway consists of two parts, one in which chains of ubiquitin are conjugated to substrate proteins, and one in which these substrate proteins are either degraded by the 26S proteasome or are de-ubiquitinated. The ubiquitin- and proteasome dependent pathway degrades a wide range of proteins in the nucleus and cytoplasm. It is highly specific, but controls a large number of cellular events due to the diversity in the conjugating enzymes. This pathway is important for removal of abnormal/damaged proteins that have had their recognition sites exposed as well as for control of specific transcription factors and cell cycle regulators. In plants, ubiquitin- and proteasome-dependent proteolysis is known to be involved in regulation of the cell cycle and transcription factors as well as endoplasmic reticulum-associated protein degradation, stress response and developmental processes, such as xylogenesis and senescence. PMID- 11473705 TI - A calcium-binding protein with similarity to serum albumin localized to the ER Golgi network and cell walls of spinach (Spinacia oleracea). AB - Using polyclonal antibodies raised against human serum albumin (HSA), a 70-kDa microsomal protein with an isoelectric point of approximately 6.5 was detected in spinach (Spinacia oleracea L.). The protein was purified by selective ammonium sulfate precipitation and anion exchange HPLC. The protein shared 100% identity with the first 15 amino acids at the NH2 terminus of HSA, including the X-X-H amino acid region, which was identified in HSA as being responsible for binding of copper, zinc, indole derivatives and calcium. Blue staining of the protein with the cationic carbocyanine dye 'Stains-all' and 45Ca overlay following SDS PAGE also suggest that the 70-kDa plant protein binds calcium. The protein reacted positively with carbohydrate specific thymol stain, and the carbohydrates associated with the protein were identified by gas chromatography-mass spectrometry (GC-MS) as galactose and galacturonic acid. The 70-kDa plant protein was present in the detergent-poor phase following Triton X-114 extraction of the microsomal proteins. Cell fractionation using continuous sucrose gradients showed that the protein is present in membrane fractions with high activity of endoplasmic reticulum (ER) and Golgi marker enzymes. Using nitrocellulose tissue prints probed with anti-HSA antibodies, we demonstrated that the protein is present in the apoplastic space of petioles, suggesting that the protein is secreted to the apoplast of cortex cells in plants. Localization and binding properties suggest that the plant protein identified in the present study may participate in secretion processes, possibly involved with the transport of precursors required for cell-wall synthesis. PMID- 11473706 TI - Grain sink strength may be related to the poor grain filling of indica-japonica rice (Oryza sativa) hybrids. AB - The physiological and biochemical factors contributing to poor grain filling of indica-japonica rice (Oryza sativa L.) hybrids were studied by analyzing the role of grain sink strength in dry matter accumulation of grains of two types of rice cultivars, Yayou 2 (an indica-japonica hybrid) and Yanjing 2 (a japonica cultivar). Carbon dioxide enrichment and plant hormone application were imposed at anthesis and the number of endosperm cells, dry matter accumulation and the activities of some sugar-metabolizing enzymes of grains were measured during grain filling. In Yayou 2, strong-potential grains (SPGs) accumulated dry weight much earlier than weak-potential grains (WPGs), but this difference was not obvious for Yanjing 2. Carbon dioxide enrichment imposed after heading significantly stimulated dry matter accumulation of WPGs of Yayou 2, but had little influence on WPGs of Yanjing 2 and SPGs of both cultivars. Leaf sheath dry matter decreased steadily in both cultivars during early stages of grain filling and accumulated during the later stages. Carbon dioxide enrichment increased leaf sheath dry matter. Dry matter accumulated by grains was linearly related to the increases in endosperm cell numbers and the activities of sucrose synthase (SS) and ADP-glucose pyrophosphorylase (AGPase) in SPGs and WPGs of both cultivars. Application of either 6-benzyladenine or abscisic acid had no significant influences on both endosperm cell number and grain dry matter accumulation. These results suggest that grain sink strength, determined by both cell numbers, SS and AGPase activities in the endosperm control the dry matter accumulation of grains. PMID- 11473707 TI - Contrasting responses of photosynthesis and carbon metabolism to low temperatures in tall fescue and clovers. AB - Growth, photosynthesis and carbohydrate metabolism in plants of two grassland species, clover (Trifolium subterraneum L. cv. Areces and Gaitan) and tall fescue (Festuca arundinacea Schreb.), shifted from 25 to 12 degrees C for 1 day or developed at 12 degrees C were compared with controls kept at 25 degrees C. Cold development produced a larger inhibition of growth in fescue than in clovers. In contrast, transferring plants from high to low temperature inhibited photosynthesis to a lesser extent in fescue than in clovers, this difference being associated with an increase in the activation state of Calvin cycle enzymes in fescue, but not in the clovers, a decreased cytosolic fructose-1,6 bisphosphatase (cFBPase, EC 3.1.3.11) activity in clovers, and an accumulation of hexose phosphates only in fescue. Development at 12 degrees C partly relieved the inhibition of photosynthesis in clovers, in contrast with fescue, which correlated with increases in total ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco, EC 4.1.1.39) activity only in clovers, and with greater increases in total stromal FBPase (sFBPase) activity in clovers than in fescue. The activity of sucrose synthesis enzymes was increased in the two clovers and fescue developed in the cold, while carbohydrate accumulation was much bigger in cold-developed fescue than in clovers because of a 5-fold increase in fructan contents in the former. The contents of phosphorylated intermediates increased in clovers but decreased in fescue grown at 12 degrees C. Our results suggest that restricted ribulose-1,5-bisphosphate (RuBP) regeneration limited the recovery of photosynthetic capacity in cold-developed fescue. PMID- 11473708 TI - Response of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii to salt-dependent oxidative stress: The root antioxidative system. AB - The response of the antioxidant system to salt stress was studied in the roots of the cultivated tomato Lycopersicon esculentum Mill. cv. M82 (Lem) and its wild salt-tolerant relative L. pennellii (Corr.) D'Arcy accession Atico (Lpa). Roots of control and salt (100 mM NaCl)-stressed plants were sampled at various times after commencement of salinization. A gradual increase in the membrane lipid peroxidation in salt-stressed root of Lem was accompanied with decreased activities of the antioxidant enzymes: superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11) and decreased contents of the antioxidants ascorbate and glutathione and their redox states. In contrast, increased activities of the SOD, CAT, APX, monodehydroascorbate reductase (MDHAR; EC 1.6.5.4), and increased contents of the reduced forms of ascorbate and glutathione and their redox states were found in salt-stressed roots of Lpa, in which the level of membrane lipid peroxidation remained unchanged. It seems that the better protection of Lpa roots from salt induced oxidative damage results, at least partially, from the increased activity of their antioxidative system. PMID- 11473709 TI - Tolerance of coffee (Coffea spp.) seeds to ultra-low temperature exposure in relation to calorimetric properties of tissue water, lipid composition, and cooling procedure. AB - The effect of exposure to ultra-low temperature (liquid nitrogen, LN) on viability of seeds desiccated to various water contents was investigated in 9 coffee species. Three groups of species could be distinguished based on seed survival after LN exposure. In group 1 species, no seedling production could be obtained after LN exposure due to endosperm injury. In group 2 species, recovery was very low or nil after rapid cooling, and only moderate after slow cooling. In group 3 species, very high percentages of seedling development were observed after both rapid and slow cooling. A high interspecific variability for the high moisture freezing limit was observed within the species of groups 2 and 3, since it ranged from 0.14 to 0.26 g H2O g-1 dry weight. A very highly significant correlation was found for those species between the unfreezable water content, as determined from DSC analysis, and the high moisture freezing limit of their seeds. No significant correlation was found between seed lipid content, which varied from 9.8 to 34.6% dry weight, and survival after LN exposure. However, a negative relationship was found between seed unfreezable water content and lipid content. Interspecific differences in fatty acid composition of seed lipids resulted in a high variability in the percentage of unsaturated fatty acids, which ranged from 28.7 to 54.4% among the 9 species studied. For all species studied, a highly significant correlation was found between the percentage of unsaturated fatty acids and the percentage of seedling recovery after rapid or slow cooling. PMID- 11473710 TI - The effect of high temperature and high atmospheric CO2 on carbohydrate changes in bell pepper (Capsicum annuum) pollen in relation to its germination. AB - Pollen viability and germination are known to be sensitive to high temperature (HT). However, the mode by which high temperature impairs pollen functioning is not yet clear. In the present study, we investigated the effect of high temperature on changes occurring in carbohydrate of bell pepper (Capsicum annuum L. cv. Mazurka) pollen in order to find possible relations between these changes and pollen germination under heat stress. When pepper plants were maintained under a moderate HT regime (32/26 degrees C, day/night) for 8 days before flowers have reached anthesis, pollen count at anthesis was similar to that found in plants grown under normal temperatures (NT 28/22 degrees C). However, the in vitro germination, carried out at 25 degrees C, of pollen from HT plants was greatly reduced. This effect matched the marked reduction in the number of seeds per fruit in the HT plants. Maintaining the plants at high air CO2 concentration (800 &mgr;mol mol-1 air) in both temperature treatments did not affect the in vitro germination of pollen from NT plants, but restored germination to near the normal level in pollen from HT plants. Under NT conditions, starch, which was negligible in pollen at meiosis (8 days before anthesis, A-8) started to accumulate at A-4 and continued to accumulate until A-2. From that stage until anthesis, starch was rapidly degraded. On the other hand, sucrose concentration rose from stage A-4 until anthesis. Acid invertase (EC 3.2.1.26) activity rose parallel with the increase of sucrose. In pollen from HT plants, sucrose and starch concentrations were significantly higher at A-1 pollen than in that of NT plants. Under high CO2 conditions, the sucrose concentration in the pollen of HT plants was reduced to levels similar to those in NT pollen. In accordance with the higher sucrose concentration in HT pollen, the acid invertase activity in these pollen grains was lower than in NT pollen. The results suggest that the higher concentrations of sucrose and starch in the pollen grains of HT plants may result from reduction in their metabolism under heat stress. Elevated CO2 concentration, presumably by increasing assimilate availability to the pollen grain, may alleviate the inhibition of sucrose and starch metabolism, thereby increasing their utilization for pollen germination under the HT stress. Acid invertase may have a regulatory role in this system. PMID- 11473711 TI - Response to mild water stress in transgenic Pssu-ipt tobacco. AB - The response of antioxidant enzymes to cyclic drought was studied in control non transformed tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) and two types of transgenic Pssu-ipt tobacco (grafted on wild rootstock and poorly rooted progeny of F1 generation) grown under different conditions of irradiation (greenhouse, referred as high light, versus growth chamber, referred as low light). Water stress cycles started with plants at two contrasting developmental stages, i.e., at the stage of vegetative growth (young) and at the onset of flowering (old). Drought reduced the growth of SR1 plants compared with transgenic ones, particularly, when treatment started in earlier stage of plant development. Relative leaf water content was significantly lower (below 70%) in all transgenic grafts and plants compared with the wild type, irrespective of age, drought, and growth conditions. The response of antioxidant enzymes was significantly dependent on plant type and plant age; nevertheless, growth conditions and water stress also affected enzyme activities. Contrary to non-transgenic tobacco, where about half of glutathione reductase activity was found in older plants, both transgenic types exhibited unchanged activities throughout plant development and stress treatment. No differences were found in catalase activity, although the growth in the greenhouse caused a moderate increase in all older plants. In contrast to non-transgenic and Pssu-ipt rooted plants, peroxidase activities (ascorbate, guaiacol, and syringaldazine peroxidase) in older Pssu-ipt grafts were up to four times higher, irrespective of growth and stress, nevertheless, the effect seemed to be age-dependent. Superoxide dismutase (SOD) activity was affected particularly by plant age but also by growth conditions. Unlike in older plants, water stress caused an increase of SOD activities in all younger plants. The differences observed in activities of enzymes of intermediary metabolism (i.e., malic enzyme and glucose-6-phosphate dehydrogenase) revealed that transgenic grafts probably compensated differently for a decrease of ATP and NADPH than control and transgenic rooted plants under stress. PMID- 11473712 TI - Regulation by light and metabolites of ferredoxin-dependent glutamate synthase in maize. AB - The regulation of Fd-glutamate synthase (Fd-GOGAT, EC 1.4.1.7) and NADH-glutamate synthase (NADH-GOGAT, EC 1.4.1.14) was investigated in maize (Zea mays L. cv. DEA) (1) during development starting from 7- to 11-day-old seedlings, (2) by treatment of 7-day-old etiolated leaves with intermittent light pulses to activate (red) and inactivate (far-red) phytochromes and (3) in 7-day-old green leaves grown under 16-h light/8-h dark cycles. Fd-GOGAT mRNA accumulated 4-fold, and the enzyme polypeptide (3-fold) and activity (3-fold) also increased in leaf cells, while NADH-GOGAT activity remained constantly low. Leaf-specific induction of Fd-GOGAT mRNA (3-fold) occurred in etiolated leaves by low fluence red light, and far-red light reversibly repressed the mRNA accumulation. Red/far-red reversible induction also occurred for Fd-GOGAT polypeptide (2-fold) and activity (2-fold), implicating the phytochrome-dependent induction of Fd-GOGAT. In contrast, NADH-GOGAT activity remained constant, irrespective of red/far-red light treatments. Fd-GOGAT showed diurnal changes under light/dark cycles with the maximum early in the morning and the minimum in the afternoon at the levels of mRNA, enzyme polypeptide and activity. Gln diurnally changed in parallel with Fd-GOGAT mRNA. The induction of Fd-GOGAT provides evidence that light and metabolites are the major signal for the Gln and Glu formation in maize leaf cells. PMID- 11473713 TI - Two distinct mechanisms regulate the transcription of photosystem II genes in Synechocystis sp. PCC 6803. AB - Expression and regulation of psb genes, encoding various subunits of photosystem II (PSII), were studied in the cyanobacterium Synechocystis sp. PCC 6803. Transcription of the psbA and psbD genes, encoding the PSII reaction centre proteins D1 and D2, was rapidly activated upon onset of illumination and the transcription rates were enhanced at high irradiance. Gel retardation analysis demonstrated dark-enhanced binding of proteins to the upstream region of the psbA2 gene, pointing to a repressor-protein-based transcriptional regulation mechanism. Transcription of all the other psb genes also required light, but unlike the psbA and psbD genes, these psb genes did not respond specifically to high-light. Moreover, the transcription of these psb genes was activated slowly at onset of illumination, and was strictly dependent on de novo protein synthesis. We suggest that these psb genes are up-regulated in the light via transcriptional activator proteins, and the slow activation may be related to production of new PSII centres during growth. Apart from the two distinct mechanisms for transcriptional regulation, all psb genes shared a common regulation mechanism at the level of transcript stability, mediated by the redox poise of intersystem electron carrier(s). PMID- 11473714 TI - Glyphosate selection of gene amplification in suspension cultures of 3 plant species. AB - Stepwise selection was carried out with increasing glyphosate concentrations to produce suspension cultures of Medicago sativa L. (alfalfa), Glycine max L. (Merr.) (soybean) and Nicotiana tabacum L. (tobacco) (two lines) that were at least 100-fold more resistant than the original culture as measured by the I50. The selection process required from 8 to 11 transfers to fresh medium over a total period from 161 to 312 days. The alfalfa and soybean lines contained 62- and 21-fold higher activity levels of the glyphosate target enzyme, 5 enolpyruvylshikimate-3-phosphate synthase (EPSPS), respectively. The tobacco lines had EPSPS enzyme activity levels more than 800-times higher than the original cultures. The EPSPS gene copy number and mRNA were increased in all of the lines as measured by southern and northern hybridization, respectively. Thus, as has been found before with most glyphosate-resistant suspension cultures, the resistance is caused by high EPSPS enzyme activity due to EPSPS gene amplification. Alfalfa and soybean EPSPS gene amplification and the very high EPSPS enzyme activity increases found in the tobacco cultures have not been reported before. These studies show that EPSPS gene amplification can occur in many plant species to confer glyphosate tolerance. PMID- 11473715 TI - Cloning and sequence analysis of cDNA encoding thiamin-binding proteins from sesame seeds. AB - The amino acid sequences of the large polypeptides of thiamin-binding proteins (TBPs) from sesame (Sesamum indicum L.) seeds (STBP-I, -II and -III) were analyzed. The large polypeptides of STBP-I, -II and -III had the same amino acid sequences as did their small polypeptides. The peptide sequence information obtained from STBPs was used to synthesize DNA primers for amplification of the gene(s) encoding STBPs. A 200-bp fragment was amplified from cDNA synthesized from RNA from sesame seeds 4 weeks after flowering. The 200-bp fragment was used to clone full-length cDNA(s) encoding STBP(s) with RACE techniques. A 644-bp fragment was amplified, cloned and sequenced. The cDNA was a full-length clone encoding STBP(s). It contained an open reading frame, which defined a 143-residue polypeptide. The identified small and large polypeptide sequences of STBPs exactly matched the sequence encoded within the cDNA clone. These results indicated that the small and large polypeptides of STBPs were encoded on the mRNA as a single large proprotein precursor and that the final mature forms were generated by post-translational processing in the same manner as the other 2S albumins of plant seeds. PMID- 11473716 TI - Polyphenol oxidase and herbivore defense in trembling aspen (Populus tremuloides): cDNA cloning, expression, and potential substrates. AB - The biochemical anti-herbivore defense of trembling aspen (Populus tremuloides Michx.) was investigated in a molecular analysis of polyphenol oxidase (PPO; EC 1.10.3.2). A PPO cDNA was isolated from a trembling aspen wounded leaf cDNA library and its nucleotide sequence determined. Southern analysis indicated the presence of two PPO genes in the trembling aspen genome. Expression of PPO was found to be induced after herbivory by forest tent caterpillar, by wounding, and by methyl jasmonate treatment. Wound induction was systemic, and occurred in unwounded leaves on wounded plants. This pattern of expression is consistent with a role of this enzyme in insect defense. A search for potential PPO substrates in ethanolic aspen leaf extracts using electron spin resonance (ESR) found no pre existing diphenolic compounds. However, following a brief delay and several additions of oxygen, an ESR signal specific for catechol was detected. The source of this catechol was most likely the aspen phenolic glycosides tremulacin or salicortin which decomposed during ESR experiments. This was subsequently confirmed in experiments using pure salicortin. PMID- 11473717 TI - Developmental expression of the mitochondrial pyruvate dehydrogenase complex in pea (Pisum sativum) seedlings. AB - In order to better understand control of the mitochondrial pyruvate dehydrogenase complex (PDC), total catalytic activity was determined during development of the primary leaves of pea (Pisum sativum L.) seedlings, as well as in each leaf pair of 21-day-old plants. Activity of the PDC in clarified homogenates was highest in the youngest organs and then dropped dramatically as the leaves matured and became photosynthetically competent. As leaves began to senesce, total PDC activity dropped to zero. Steady-state mRNA levels were determined using E1 and E3 cDNA probes. The overall pattern of transcript abundance matched the pattern observed for total PDC activity; transcript levels for E1alpha and E1beta approached zero during senescence. Levels of the E1alpha, E1beta, E2 and E3 subunits of the PDC were analyzed in the same samples, using specific antibodies. Quantitation of the immunoblotting results throughout this developmental series showed a pattern in parallel with that of catalytic activity and mRNA levels, although the relative changes in subunit protein levels were not as extreme as the changes in activity. The exception to the global pattern was that of the E3 subunit: lipoamide dehydrogenase. Expression of this enzyme was highest in mature, fully expanded leaves, which were active in photosynthesis and photorespiration, reflecting the additional role of E3 as a component of glycine decarboxylase. PMID- 11473718 TI - Effect of the tetrapeptide RGDS on somatic embryogenesis in Daucus carota. AB - The present work examines the requirement for RGD-binding sites, such as those in the plasma membrane protein integrin during somatic embryogenesis in carrot (Daucus carota L. cv. Scarlet nantes). It is possible to assay for this requirement by competitively inhibiting binding of the site to the natural RGD containing ligand by adding an excess of synthetic RGDS. We found that treatment of carrot callus cultures with RGDS (2.7 mM) inhibited the development of a normal shoot pole in carrot somatic embryos. The structures that formed contained separate zones of highly cytoplasmic and vacuolate cells and no evidence of embryonic organization, although occasionally a root-like structure was observed. If the aspartic acid residue in the peptide was replaced by a chemically similar amino acid (glutamic acid), the resultant somatic embryos were indistinguishable from those developing in untreated cultures. These effects are similar to those reported in animal systems where the protein receptor involved has been identified as integrin. Our results are thus consistent with the conclusion that a binding site for a motif similar to the integrin-binding site in a variety of animal extracellular matrix proteins exists in plants and appears to be important in somatic embryo development in carrot. PMID- 11473719 TI - Two forms of phycobilisomes in the Antarctic red macroalga Palmaria decipiens (Palmariales, Florideophyceae). AB - The phycobilisomes (PBS), the light-harvesting antennae, from the endemic Antarctic red macroalga Palmaria decipiens were isolated on discontinuous sucrose gradients in two discrete bands and not in one as expected. To exclude methodical faults, we also isolated PBS from the temperate Palmaria palmata and the unicellular red algae Porphyridium cruentum and Rhodella violacea. In P. palmata the PBS were separated in two discrete bands, whereas the PBS from Porphyridium and Rhodella were found in one band. The double-banded PBS (PBSup and PBSlow) from P. decipiens were further characterized by absorption and fluorescence spectroscopy, native and SDS-PAGE as well as by negative staining. The phycobiliproteins RIII-phycoerythrin, RI-phycocyanin and allophycocyanin were identified and 3 gamma-subunits were described. The PBSup and PBSlow showed no significant differences in their absorption spectra and phycobiliprotein ratios although the negative stained PBSlow were smaller. Differences were found in their low molecular mass subunit complexes, which are assumed to be r phycoerythrin. The polypeptide pattern of the PBSup and PBSlow showed no differences in the molecular masses of their subunits and linker polypeptides, but in their percentage distribution. The results suggest that the PBSlow is a closer packed and PBSup a little more loosely aggregated hemiellipsiodal PBS form. We discuss the ecophysiological function of two PBS forms in P. decipiens and suggest advantages in the rapid acclimation to changes in environmental light conditions. PMID- 11473720 TI - Consequence of restricted mitochondrial oxidative metabolism on photosynthetic carbon assimilation in mesophyll protoplasts: Decrease in light activation of four chloroplastic enzymes. AB - The patterns of light activation of 4 chloroplastic enzymes were examined in mesophyll protoplasts of pea (Pisum sativum) in the absence or presence of oligomycin (inhibitor of oxidative phosphorylation) or antimycin A (inhibitor of cytochrome pathway) or salicylhydroxamic acid (SHAM, inhibitor of alternative pathway). The results were compared with those of DCMU (inhibitor of photosynthetic electron transport). The light activation of NADP glyceraldehyde-3 phosphate dehydrogenase (NADP-GAPDH), fructose-1,6-bisphosphatase (FBPase), phosphoribulokinase (PRK) (enzymes of the Calvin cycle) and NADP malate dehydrogenase (NADP-MDH) (reflects chloroplast redox state) was more pronounced at limiting CO2 (0.1 mM NaHCO3) than that at optimal CO2 (1.0 mM NaHCO3). SHAM decreased markedly (up to 33%) the light activation of all 4 enzymes, while antimycin A or oligomycin exerted only a limited effect (<10% decrease). Antimycin A or oligomycin or SHAM had no significant effect on light activation of these 4 enzymes in isolated chloroplasts. However, DCMU caused a remarkable decrease in light activation of enzymes in both protoplasts (up to 78%) and chloroplasts (up to 69%). These results suggest that the restriction of alternative pathway of mitochondrial metabolism results in a marked decrease in the light activation of key chloroplastic enzymes in mesophyll protoplasts but not in isolated chloroplasts. Such a decrease in the light activation of enzymes could be also a secondary feedback effect because of the restriction on carbon assimilation. PMID- 11473721 TI - Contribution to promoting cancer epidemiology in Japan and to the activities of the UICC and others in cancer epidemiology and prevention. AB - On the occasion of being awarding the 5th Nagayo Mataro Prize for contributions to promoting cancer epidemiology and international involvement in the UICC (International Union against Cancer), the Monbusho Overseas Field Research (International Scientific Research)-Special Cancer Study and others, it is my pleasure to give a concise description of activities in which I have played a role. My achievements in administration and management are only a small part of the whole of what has been accomplished in cancer epidemiology and prevention, but I hope that a comprehensive coverage of the projects in which it has been my good fortune to participate may provide orientation and suggest priorities for research in the coming decades. I have taken part in many meetings that stimulated the interest of young scientists and physicians in epidemiology and prevention. It can be said that efficient administration and management are indispensable for modern scientific research in order to promote information collection and exchange. I, therefore, humbly hope that my experience may be illuminating. PMID- 11473722 TI - Preventive effects of isoflavones, genistein and daidzein, on estradiol-17beta related endometrial carcinogenesis in mice. AB - The effects of isoflavones (genistein and daidzein) on endometrial carcinogenesis in mice were investigated in two experiments. In the short-term experiment (2 weeks), single subcutaneous (s.c.) administration of genistein [1 mg / 30 g body weight (b.w.)] significantly decreased the levels of estradiol-17beta (E(2)) (5 ppm in diet)-induced expression of c-jun, interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha) mRNAs in the uteri of ovariectomized mice (P < 0.005, P < 0.05 and P < 0.01, respectively). Daidzein significantly inhibited E(2)-induced expression of c-fos and IL-1alpha (P < 0.01, P < 0.01 respectively). In the long-term experiment (30 weeks), 140 female ICR mice were given N-methyl-N-nitrosourea-containing solution (1 mg / 100 g b.w.) and normal saline (as controls) into their left and right uterine corpora, respectively. They were divided into six groups; group 1 was given E(2) (in diet) alone. Group 2 was given E(2) and genistein (1 mg / 30 g b.w., s.c., every four weeks). Group 3 was exposed to E(2) and daidzein (1 mg / 30 g b.w., s.c., every four weeks). Groups 4 and 5 respectively received genistein and daidzein, and were kept on the basal diet. Group 6 was kept on the basal diet and served as a control. At the termination of the experiment, incidences of endometrial adenocarcinoma and atypical endometrial hyperplasia of the group given E(2) and genistein or daidzein were significantly lower than of the group with E(2) alone (P < 0.01 and P < 0.05, respectively). It is suggested that both genistein and daidzein have an inhibitory effect on estrogen-related endometrial carcinogenesis in mice, possibly by suppressing expression of estrogen-induced estrogen-related genes c fos and c-jun, and internal cytokines IL-1alpha and TNF-alpha through a cytokine and estrogen receptor-mediated pathway. PMID- 11473723 TI - Different susceptibility of each L-myc genotype to esophageal cancer risk factors. AB - To understand the relationship between the L-myc genotypes and esophageal cancer risk, a polymerase chain reaction-based restriction fragment length polymorphism analysis was performed on 91 Japanese patients with esophageal cancer and 241 non cancer outpatients. No significant difference in the distribution of genotypes was observed between patients and controls; 18.7% LL genotype, 56.0% LS and 25.3% SS among patients, and 24.5%, 55.6% and 19.9%, respectively, among controls. Frequency of the s-allele in patients (0.533) was slightly higher than in controls (0.477), but the difference was not statistically significant. However, the odds ratios (ORs) for smoking or heavy drinking were markedly higher in SS and LS genotypes than in LL genotype; age-sex-adjusted ORs for smoking was 7.57 in the SS genotype, 6.40 in the LS genotype and 1.77 in the LL genotype. Age-sex adjusted ORs for heavy drinking were 19.78, 18.20 and 7.40, respectively. The age sex-adjusted ORs for both factors combined were 12.77, 18.45 and 1.44, respectively. These results suggested that the L-myc polymorphism might modify the effects of lifestyle factors on esophageal cancer risk. PMID- 11473724 TI - DNA sequence copy number aberrations associated with histological subtypes and DNA ploidy in gastric carcinoma. AB - We have analyzed DNA sequence copy number aberrations (DSCNAs) and DNA ploidy by using comparative genomic hybridization and laser scanning cytometer in gastric carcinomas (GCs) to elucidate the genomic aberrations in relation to clinicopathological parameters. Thirty-two out of 33 cases showed one or more DSCNAs with a mean number of 11.7 per tumor. High-level gains were detected at 2p, 3q, 6p, 7p, 7q, 8q, 12p, 13q, 19q, and 20q. Frequency of gross genomic abnormalities and chromosome regions that have genomic aberrations were similar in both intestinal- and diffuse-type GCs, except aberrations at 8p, 9p, 12q, and 20q. The overall number of DSCNAs was significantly greater in DNA aneuploid tumors than that in DNA diploid tumors. We detected genomic aberrations characterized by histological subtype, tumor location, and DNA ploidy status: gain of 20q and losses of 8p and 9p in intestinal-type GCs, gains of 8p and 12q in diffuse-type GCs, gain of 20q in the lower third GCs, and loss of 5q, 9p, 10q, 16q, and 18q in DNA aneuploid GCs. Furthermore, 5q loss is associated with DNA aneuploidy (P = 0.0001) or the total number of losses (P = 0.001), gain + losses (P = 0.004), and high-level gains (P = 0.001) in GCs. Among these loci, chromosome 8p was unique. Gain of 8p was more common in diffuse-type GC, whereas loss of 8p was more frequently detected in intestinal-type GC. In conclusion, we describe chromosomal regions of 5q, 8p, and 20q, which are of interest for further investigation of GCs. PMID- 11473725 TI - A novel experimental mouse model of peritoneal dissemination of human gastric cancer cells: analysis of the mechanism of peritoneal dissemination using cDNA macroarrays. AB - We established a new cell line, NUGC-3P4T, with high peritoneal metastatic disseminating potential in nude mice. NUGC-3P4T cells were derived from the human gastric carcinoma line NUGC-3, which has low capacity for peritoneal dissemination. NUGC-3P4T cells developed peritoneal dissemination in 10 / 10 (100%) mice, whereas the parental NUGC-3 cells developed dissemination in 1 / 5 (20.0%) mice. The metastatic foci in the peritoneum showed essentially the same histological appearance as those induced by parental cells. The tumorigenicity, the motile activity and the adhesive activity to the laminin of NUGC-3P4T cells were stronger than those of NUGC-3 cells. Production of IL-8 was significantly higher in NUGC-3P4T than in NUGC-3. cDNA macroarrays analysis showed that a variety of cytokines, interleukins, and other immunomodulators and their receptors were up- or down-regulated at the mRNA level in NUGC-3P4T cells, compared with NUGC-3 cells. Thus, this unique cell line and in vivo model might be useful to study the biology of peritoneal dissemination of human gastric cancer. PMID- 11473726 TI - Phenotypic expression of colorectal adenocarcinomas with reference to tumor development and biological behavior. AB - The purpose of this study is to clarify the correlation between cell differentiation and tumor development, including tumor aggressiveness and biological behavior. Eighty-three cases of advanced colorectal adenocarcinoma were randomly selected. Using immunohistochemical staining with antibodies to CD10, MUC2 and human gastric mucin (HGM), the colorectal adenocarcinomas could be classified into five types (18 small intestinal, 27 large intestinal, 2 gastric, 9 mixed and 27 unclassified). Each type had characteristic features. The small intestinal type showed a relatively lower incidence of lymphatic permeation and higher venous invasion. The large-intestinal type showed a low incidence of venous invasion and lymph node metastasis. The mixed type revealed female and right-side-dominant distribution, large tumor size, high incidence of mucinous carcinoma, and low incidence of venous invasion. Gastric type was seen in only two cases (2%), which exhibited high histologic grade, lymphatic permeation and lymph node metastasis with no venous invasion. Such phenotypic classifications are considered to be useful not only for evaluation of the biological behavior of the carcinoma, but also for analysis of tumorigenesis. PMID- 11473727 TI - Identification of cyclophilin B-derived peptides capable of inducing histocompatibility leukocyte antigen-A2-restricted and tumor-specific cytotoxic T lymphocytes. AB - We recently suggested that cyclophilin B (Cyp-B) is a tumor antigen recognized by histocompatibility leukocyte antigen (HLA)-A24-restricted and tumor-specific cytotoxic T lymphocytes (CTLs). In this study, we tried to identify Cyp-B-derived epitopes, which can induce HLA-A2-restricted and tumor-specific CTLs in cancer patients. The tumor-infiltrating lymphocytes (TILs) from an HLA-A0207 patient with colon cancer were found to respond to COS-7 cells when co-transfected with the Cyp-B gene and either HLA-A0201, -A0206, or -A0207 cDNA. These TILs contained CTLs capable of recognizing either the Cyp-B(129 - 138) or the Cyp-B(172 - 179) peptide among 28 different peptides, all of which were prepared based on the HLA A2 binding motif. Both Cyp-B peptides possessed the ability to induce tumor specific CTLs in HLA-A2(+) cancer patients. Cyp-B(172 - 180 (V)), which is a 9 mer peptide with valine added at the C terminus, showed no clear superiority over the parental Cyp-B(172 - 179) peptide in an in vitro sensitization experiment. In vitro-sensitized T cells with these peptides responded to cancer cells in an HLA A2-restricted manner. These two Cyp-B peptides could be useful for specific immunotherapy of HLA-A2(+) cancer patients. PMID- 11473728 TI - Enhancement of in vivo antitumor activity of a novel antimitotic 1 phenylpropenone derivative, AM-132, by tumor necrosis factor-alpha or interleukin 6. AB - TK5048 and its derivatives, AM-132, AM-138, and AM-97, are recently developed antimitotic (AM) compounds. These 1-phenylpropenone derivatives induce cell cycle arrest at the G2 / M phase of the cell cycle. TK5048 inhibited tubulin polymerization in human lung cancer PC-14 cells in a concentration-dependent manner. In a polymerization assay using bovine brain tubulin, AM-132 and AM-138 were quite strong, AM-97 was moderately strong, and TK5048 was a relatively weak inhibitor of tubulin polymerization. A murine leukemia cell line resistant to a sulfonamide antimitotic agent, E7010, which binds to colchicine-binding sites on tubulin, was cross-resistant to the in vitro growth-inhibitory effect of AM compounds. Inhibition of tubulin polymerization is therefore one of the mechanisms of action of these AM compounds against tumor cells. To profile the antitumor effect of AM compounds, the in vivo antitumor effect of AM-132 was evaluated against cytokine-secreting Lewis lung carcinoma (LLC). Tumor-bearing mice were treated with intravenous AM-132 using three different treatment schedules. LLC tumors expressing tumor necrosis factor-alpha (TNF-alpha), granulocyte macrophage colony-stimulating factor (GM-CSF), or interleukin (IL)-6 were very sensitive to AM-132. In particular, LLC tumors expressing IL-6 were markedly reduced by AM-132 treatment, and showed coloring of the tumor surface and unusual hemorrhagic necrosis. These results suggest a combined effect of AM 132 and cytokines on the blood supply to tumors. PMID- 11473729 TI - Expression of drug resistance genes in VP-16 and mAMSA-selected human carcinoma cells. AB - The cell lines described in the present study were isolated as part of an effort to understand resistance to topoisomerase (topo) II inhibitors. To that end, 50 sublines were isolated from four human breast cancer cell lines, i.e., MCF-7, T47D, MDA-MB-231, and ZR-75B. As an initial step, a concentration that would be lethal to the majority of cells (IC99) was selected for both VP-16 and mAMSA, for each cell line. The identification of an increasing number of putative drug resistance-related proteins provided the opportunity to examine expression of the corresponding genes in the selected cell lines. Northern blot analysis revealed different responses to the selecting agents in the different cell lines. Previous studies examining expression of multidrug resistance (MDR)-1 in resistant cell lines had found undetectable levels in all cells. In the ZR-75B sublines, increased expression of MDR-associated protein (MRP) and canalicular multispecific organic anion transporter (cMOAT) was observed, and when the relative levels of overexpression were compared, a high correlation was found. In contrast, increased expression of MRP was observed in some of the MDA-MB-231 sublines, without a concomitant increase in cMOAT expression. Finally, in both T47D and MCF-7 sublines, increased expression of cMOAT or MRP was observed infrequently, and where it occurred, was of a much smaller magnitude. In the analysis of expression of MRP, the highest levels were found in the ZR-75B and MDA-MB-231 sublines, with lower levels in the MCF-7 and T47D clones. Similarly, differences in the expression of topo IIalpha were observed among the sublines. Although the differences in expression appear to depend on the parental cell line from which the resistant sublines were derived, a strong correlation was observed between the expression of MRP and the levels of topo IIalpha. Cell lines with low levels of MRP had lower levels of topo IIalpha, while those with high levels of MRP maintained higher levels of topo IIalpha. While a reduced topo IIalpha level was common, there did not appear to be a compensating increase in the expression of topo IIbeta or topo I or casein kinase (CK) IIalpha in any of the cell lines. While the possibility that such compensation could occur has been discussed and even reported in some cell lines, such an adaptation was not observed in the present study, suggesting that it is not common. PMID- 11473730 TI - A new quinoline derivative MS-209 reverses multidrug resistance and inhibits multiorgan metastases by P-glycoprotein-expressing human small cell lung cancer cells. AB - Development of distant metastases and acquired multidrug resistance (MDR) are major problems in therapy for human small cell lung cancer (SCLC). MS-209 is a novel quinoline compound, which reverses P-glycoprotein (P-gp)-mediated MDR. We previously reported that MS-209 reversed in vitro MDR of human SCLC (SBC-3 / ADM and H69 / VP) cells expressing P-gp. In the present study, we determined the therapeutic effect of MS-209 in combination with chemotherapy against multiorgan metastases of MDR SCLC cells. SBC-3 / ADM cells expressing P-gp were highly resistant to etoposide (VP-16), adriamycin (ADM), and vincristine (VCR) in vitro, compared with parental SBC-3 cells lacking P-gp expression. MS-209 restored chemosensitivity of SBC-3 / ADM cells to VP-16, ADM, and VCR in a dose-dependent manner in vitro. Intravenous injection with SBC-3 or SBC-3 / ADM cells produced metastatic colonies in the liver, kidneys and lymph nodes in natural killer (NK) cell-depleted severe combined immunodeficiency (SCID) mice, though SBC-3 / ADM cells more rapidly produced metastases than did SBC-3 cells. Treatment with VP-16 and ADM reduced metastasis formation by SBC-3 cells, whereas the same treatment did not affect metastasis by SBC-3 / ADM cells. Although MS-209 alone had no effect on metastasis by SBC-3 or SBC-3 / ADM cells, combined use of MS-209 with VP-16 or ADM resulted in marked inhibition of metastasis formation by SBC-3 / ADM cells to multiple organs. These findings suggest that MS-209 reversed the MDR of SBC-3 / ADM cells, but not SBC-3 cells, growing in the various organs, and inhibited metastasis formation in vivo. Therefore, this chemosensitizing agent, MS-209, may be useful for treatment of refractory SCLC patients with multiorgan metastases. PMID- 11473731 TI - Deletion mutants of human deoxycytidine kinase mRNA in cells resistant to antitumor cytosine nucleosides. AB - We studied mutational events in deoxycytidine (dCyd) kinase mRNA expression, focusing on aberrant dCyd kinase mRNA, which has been frequently observed in established cell lines resistant to antitumor dCyd nucleoside analogues such as 1 beta-D-arabinofuranosyl cytosine (Ara-C), gemcitabine (dFdC) and 2'-C-cyano-2' deoxy-1-beta-D-arabinofuranosylcytosine (CNDAC). We describe here the expression of aberrant dCyd kinase mRNAs identified as splicing mutants. These mutants included deletions of the fifth exon in CNDAC-resistant cells (originating from HT-1080 cells), of the third exon in Ara-C-resistant cells (originating from SK MEL-28 cells) and of the fourth exon in 2'-deoxy-2'-methylidenecytidine (DMDC) resistant cells (originating from SK-MEL-28 cells). Various nucleoside-resistant cells originating from the same parental HT-1080 cells were established. The resulting cells expressed the same mRNA with deletion of the fifth exon, and the location of splicing was independent of the type of nucleosides used for the establishment of resistant cells. The deletion of the fifth exon in dCyd kinase seems to be a target for acquisition of resistance to antitumor cytosine nucleosides. However, distinct mutations in the dCyd kinase gene seem to be associated with acquisition of resistance to different antitumor cytosine nucleosides. PMID- 11473732 TI - Hypophosphorylation of topoisomerase IIalpha in etoposide (VP-16)-resistant human carcinoma cell lines associated with carboxy-terminal truncation. AB - Topoisomerase IIalpha is a target for many chemotherapeutic agents in clinical use. To define mechanisms of resistance and regions crucial for the function of topoisomerase IIalpha, drug-resistant cell lines have been isolated following exposure to topoisomerase II poisons. Two resistant sublines, T47D-VP and MCF-7 VP, were isolated from human carcinoma cell lines following exposure to 300 or 500 ng / ml etoposide (VP-16). Cytotoxicity studies confirmed resistance to etoposide and other topoisomerase II poisons. KCl-sodium dodecyl sulfate (K-SDS) precipitation assays using intact cells showed reduced DNA-topoisomerase II complex formation following VP-16 or amsacrine (m-AMSA). RNAse protection analysis identified a deletion of 200 base pairs in the topoisomerase IIalpha cDNA of T47D-VP and rising dbl quote, left (low)AA insertion" in the topoisomerase IIalpha cDNA of MCF-7-VP. Reduced topoisomerase IIalpha mRNA and protein levels were observed in both cell lines. It was somewhat surprising to find that nuclear extracts from T47D-VP and MCF-7-VP cells had comparable topoisomerase II activity to that of parental cells. Analysis of the extent of phosphorylation demonstrated that topoisomerase IIalpha from the resistant cells was relatively hypophosphorylated compared to that of parental cells. In these cell lines, hypophosphorylation secondary to loss of a portion of the C-terminal domain of topoisomerase IIalpha mediated the restored activity, despite a fall in topoisomerase IIalpha mRNA and protein, and this resulted in cross resistance to topoisomerase II poisons. PMID- 11473733 TI - Expression of ICAD-l and ICAD-S in human brain tumor and its cleavage upon activation of apoptosis by anti-Fas antibody. AB - ICAD / DFF is a downstream molecule of caspases, participating in nuclear DNA fragmentation during apoptosis. ICAD / DFF binds CAD / DFF40 and inhibits its DNase activity. ICAD / DFF has two alternative isoforms, long isoform (ICAD-L / DFF45) and short isoform (ICAD-S / DFF35). We have studied the presence and functional status of ICAD / DFF in human glioma cell lines. All cell lines tested expressed both ICAD-L and ICAD-S. When the cultured glioma cells were exposed to anti-Fas antibody, these isoforms were degraded prior to the fragmentation of the nuclear DNA, indicating that the ICAD / DFF expressed in cultured glioma cells was potentially functional. In primary brain tumors and normal brain tissues, there was a difference in the expression level between ICAD-L and ICAD-S. In glioblastomas, ICAD-S was more abundant than ICAD-L. In contrast, ICAD-L was more abundant than ICAD-S in medulloblastomas. The present findings suggest that primary brain tumors and normal brain constitutively express ICAD / DFF, and that there is a difference between the expression levels of ICAD-L and ICAD-S. PMID- 11473734 TI - A simple method for classification of cell death by use of thin layer collagen gel for the detection of apoptosis and/or necrosis after cancer chemotherapy. AB - To assess the efficacy of cancer chemotherapy, an important index is apoptosis of the target cells, which can usually be confirmed by electron microscopy (EM). We established a new experimental technique, whereby cancer cells (MKN45) were distributed in thin collagen gel as one or two cell layers, and cultured with anti-cancer drugs (5-FU and CDDP). The cells were stained with fluorescent Hoechst 33258 (Ho) and photographed, then with hematoxylin and eosin (H&E) and again photographed, and processed for EM. This approach allowed us to characterize the patterns of death of single cells in detail. There were six patterns of cell damage: two patterns of apoptosis, early peripheral condensation of chromatin and late apoptotic bodies, two patterns of necrosis, cytoplasmic swelling and washed-out images, and two further patterns, with morphological features of both apoptosis and necrosis, neither classified into necrosis nor apoptosis. The results show that cell death patterns can be mostly determined by combining observations of Ho and H&E-stained cells without the necessity for EM observation. PMID- 11473735 TI - First International Meeting of the Society of Atherosclerosis Imaging. February 2 4, 2001. San Diego, California, USA. Proceedings and abstracts. PMID- 11473736 TI - The atherosclerosis epidemic: methodology, nosology, and clinical practice. AB - An electronic search shows that there is a small but respectable amount of research on psychiatric and neurologic nosology. There is a also a small body of literature on endocrinologic nosology, but cardiologists almost never write about the nosology of heart disease. It does not keep them awake at night. Our understanding of the 20th-century epidemic of coronary artery disease (CAD), which dominates cardiology, has nevertheless been frustrated by problems of nosology. One of the purposes of this article is to discuss how our understanding of the epidemic has depended on classification of disease events and on the methods used to detect them. The other purpose is to show how current reclassifications of CAD and the major metabolic condition conducive to CAD, type 2 diabetes, each affect the practice of cardiology and medicine. PMID- 11473737 TI - Coronary plaque as a replacement for age as a risk factor in global risk assessment. AB - Risk assessment is assuming an increasing role for identification of high-risk persons for intensive medical intervention to reduce risk for coronary heart disease (CHD). Of particular importance is the need to identify those persons with CHD risk equivalents who can be managed with the same intensity as patients with established CHD. For example, the National Cholesterol Education Program (NCEP) recently classified diabetes as a CHD risk equivalent. The NCEP also recommended use of Framingham risk scoring in persons with multiple (2+) risk factors to uncover others without diabetes who have CHD risk equivalents. One limitation of Framingham risk scoring, however, is that age becomes the dominant risk factor after age 50. Age is a surrogate for coronary atherosclerotic plaque burden, which is the true risk factor. However, for individuals, coronary plaque burden can vary greatly at any given age. For this reason, if coronary plaque burden could be measured accurately with noninvasive techniques, the degree of plaque burden could be used to replace age as a risk factor in Framingham scoring for risk prediction. This article describes a technique whereby such a replacement can be made. PMID- 11473738 TI - The inflamed plaque: cytokine production and cellular cholesterol balance in the vessel wall. AB - Although the concept that inflammation plays a role in the biology of atherosclerosis is now well accepted, the basic feature of the arterial lesion remains the accumulation of clusters of foam cells. These clusters are the consequence of the enhanced recruitment of monocytes in the vessel wall induced by the hyperlipidemia and of the disproportionate accumulation of lipids in the cytoplasm of macrophages deriving from monocytes. Ultimately, every molecular force and pathway with modulating activity over the developing lesion will have to act on a convergence point with factors regulating cholesterol balance in the macrophage. Consistent with this view is the recent report that cytokines, such as tumor necrosis factor-alpha, can influence the expression of the scavenger receptor, whereas interferon-gamma can inhibit adenosine triphosphate-binding cassette transporter-1, the main effector of cholesterol efflux in the peripheral cell. Conversely, recent data have shown that primary alterations in macrophage cholesterol balance, such as those produced by the total absence of acylcoenzyme A:cholesterol acyltransferase-1, may determine local changes compatible with the activation of inflammatory pathways. In this brief review, we discuss some of the convergence points between inflammation and cholesterol balance, and we highlight the additional therapeutic targets suggested by these new developments in vascular biology. PMID- 11473739 TI - The development of calcium deposits in atherosclerotic lesions and their persistence after lipid regression. AB - Children have both macrophages and macrophages that are filled with lipid droplets (foam cells) at susceptible sites of arteries. Such changes are minimal and may not develop further. However, in some adolescents, small pools of dead foam cell remnants and lipid droplets (extracellular lipid) are added to the foam cell accumulations at the susceptible sites. The pools are the precursor of a much larger confluent accumulation of extracellular lipids (the lipid core)-the hallmark of the atheromas of young adults. As soon as a lesion with a lipid core is present, calcium granules of microscopic size are found among the packed extracellular particles and droplets and in smooth-muscle cells isolated among them. Disintegration of arterial structure at the core facilitates tears at the surface, hematoma, and thrombosis. As a response, layers of reparative fibromuscular tissue are added and may predominate in a lesion. Over time, calcium lumps and plates form through accretion of adjacent extracellular calcium granules. In adults past the fourth decade of life, the greater part of the former lipid core of a lesion may be calcified and there may be osseous metaplasia. The effect of therapeutic reduction of high levels of blood cholesterol on lesions was studied in rhesus monkeys. Drastic reduction of blood cholesterol levels for 312 years resulted in the disappearance of macrophages, macrophage foam cells and lymphocytes, and reduction of extracellular lipid from advanced lesions. Calcium deposits remained in the arterial wall and were not visibly changed. PMID- 11473740 TI - Insights into the mechanism of vascular calcification. AB - Vascular calcification is common and clinically significant in atherosclerosis and heart failure. It was long believed to be an end-stage process of "passive" mineral precipitation. However, there is now a growing awareness that vascular calcification is a biologically regulated phenomenon. It has many similarities to bone formation, and ectopic bone is a well-documented part of vascular calcification. This implies that alterations in vascular cell differentiation, extensive or localized, are an integral part of vascular calcification. Matrix gamma-carboxylated glutamate (GLA) protein (MGP)-deficient mice develop extensive vascular calcification with replacement of the media by progressively calcifying cartilage. A potential mechanism that explains these findings is MGP interference with bone morphogenetic proteins-potent inducers of cartilage and bone. PMID- 11473741 TI - Electron-beam tomography coronary calcium scores are superior to Framingham risk variables for predicting the measured proximal stenosis burden. AB - Previous studies of electron-beam tomography (EBT) have correlated coronary calcium scores with simplistic visual estimates of disease severity. In a clinical trial designed to evaluate 2 treatment strategies in coronary artery disease (CAD) patients with low levels of high-density lipoprotein cholesterol, we used quantitative coronary angiography to measure composite proximal stenosis burden from the baseline coronary angiogram and assessed the traditional Framingham risk variables in 146 patients. Stenosis burden is the sum, per patient, of percent stenosis for the worst lesion found in each of 9 standard proximal coronary segments. EBT estimates of coronary calcium (Agatston score, calcium volume score) were obtained for 115 of these patients. Stenosis burden was correlated with the calcium scores and risk variables. The best traditional correlates of stenosis burden were smoking status (r = 0.31, p = 0.001), prior myocardial infarction (r = 0.24, p = 0.005), body mass index (r = 0.23, p = 0.005), pack-years smoking (r = 0.20, p = 0.05), and age (r = 0.17, p = 0.04). With adjustment for age, all these correlations improved (eg, body mass index x age [r = 0.28, p = 0.001]). In addition, total cholesterol x age (r = 0.22, p = 0.008), fibrinogen x age (r = 0.19, p = 0.03), and systolic blood pressure x age (r = 0.18, p = 0.03) became significant correlates. Spearman correlations of the calcium scores with stenosis burden were considerably greater (Agatston: r = 0.62, p <0.0001; calcium volume: r = 0.63, p <0.0001). In multivariate regression analysis, calcium score, body mass index, and history of myocardial infarction were independent correlates of stenosis burden (R(2) = 0.45). At a given point in time, the EBT coronary calcium scores are greatly superior to the Framingham risk factors in predicting the measured proximal stenosis burden. Agatston and calcium volume scores are comparably predictive of stenosis burden. PMID- 11473742 TI - Predictive value of carotid 2-dimensional ultrasound. AB - Several pieces of evidence provide a rationale for an association between disease of the extracranial carotid arteries and incident coronary artery disease (CAD): (1) patients with transient ischemic attack are most likely to die from CAD; (2) atherosclerosis of the extracranial carotid arteries is correlated with that of the coronary arteries; (3) stenosis of the extracranial carotid arteries is associated with incident CAD; (4) risk factors for extracranial carotid atherosclerosis are also risk factors for CAD; and (5) there is an association between wall thickness of the extracranial carotid arteries (extracranial intimal medial thickness [IMT]) and prevalent CAD, as well as CAD and stroke. Accordingly, large population-based studies have demonstrated an association between IMT and incident CAD and stroke in younger (Atherosclerosis Risk in Communities study, 45 to 65 years of age) as well as older (Cardiovascular Health Study, > or =65 years of age) samples. IMT, measured at 1 point in time, is likely to be an excellent reflection of an individual's past exposure to risk factors. However, a single measure of IMT might bear an imperfect relation to incident events, because current risk may be influenced more by current risk factor burden than by past exposure. Longitudinal studies have shown an association of risk factors with IMT progression, and clinical trials have demonstrated that lipid-lowering therapy retards the rate of progression of disease. In addition, IMT progression has been shown to correlate with incident CAD. We therefore suggest that the best index of future CAD risk may be progression of IMT rather than IMT itself. PMID- 11473743 TI - Measurement of endothelial function by brachial artery flow-mediated vasodilation. AB - In response to physical and chemical stimuli, the endothelium regulates vascular tone through the release of vasodilators, such as nitric oxide, and vasoconstrictors, such as endothelin. Increased blood flow is an important stimulus for endothelium-mediated vasodilation (flow-mediated vasodilation). Brachial artery flow-mediated vasodilation can be assessed using high-frequency ultrasound assessment of changes in brachial artery diameter after a 5-minute blood pressure cuff arterial occlusion. Endothelial dysfunction is demonstrated as reduced vasodilation, which is usually associated with the presence of coronary risk factors. Brachial artery flow-mediated vasodilation correlates with coronary artery flow-mediated vasodilation. Both indexes have been shown to predict long-term cardiovascular events. Whereas the clinical applicability of brachial artery flow-mediated vasodilation is unclear, it has become a frequently employed measure of endothelial function. PMID- 11473744 TI - Current status of mechanical computed tomography in cardiac imaging. AB - Mechanical computed tomography (CT) scanners have undergone significant improvements in their ability to perform cardiac imaging. Previously limited by cardiac motion artifacts and slow acquisition speeds, newer-generation scanners are able to obtain multiple slices with a temporal resolution of <100 msec, minimizing motion artifacts to levels comparable to electron-beam computed tomography (EBCT). This is done using various approaches, including (1) prospective and retrospective electrocardiographic gating, (2) multislice detectors, and (3) partial and segmented reconstruction algorithms. The major CT manufacturers are putting significant resources into protocol optimization and cardiac applications development, including coronary calcium quantification, functional analysis, and coronary CT angiography. Given the rapid pace of development, higher in-plane and z-axis spatial resolution, and superior signal to-noise ratio of mechanical scanners, it is evident that they will be capable of performing cardiac imaging with a quality approaching or possibly surpassing that of existing EBCT scanners. This article summarizes the current status of mechanical CT scanners in cardiac imaging. PMID- 11473745 TI - Electron-beam computed tomography: a Bayesian approach to risk assessment. AB - The epidemic of coronary artery disease continues to affect a large number of individuals who often experience sudden and unexpected events. This underscores the need to develop more effective programs to detect silent atherosclerosis, with the ultimate goal of preventing coronary events. The use of conventional risk factors is helpful in assessing the median risk of a population, but it is often unsatisfactory in estimating the actual risk of an individual patient. As a consequence, newer imaging modalities are being developed to detect atherosclerosis in its early developmental phases. Technologies such as electron beam computed tomography (EBCT) may render risk stratification more accurate if used in the appropriate patient populations and with the right diagnostic approach. Several studies have already demonstrated the power of coronary calcification as a strong predictor of future cardiovascular events. Nonetheless, the medical literature is currently pervaded by an animated debate, as some investigators still have concerns about the effectiveness of a preventive approach driven by technology. The use of Bayesian models to interpret data acquired with EBCT screening may provide practitioners with valuable evidence to aid in their decision making. PMID- 11473746 TI - The human high-risk plaque and its detection by magnetic resonance imaging. AB - The study of atherosclerotic disease during its natural history and after therapeutic intervention will enhance our understanding of the progression and regression of this disease and will aid in selecting the appropriate treatments. Several invasive and noninvasive imaging techniques are available to assess vessels in atherosclerotic disease. Most of the standard techniques, however, identify luminal diameter or stenosis, wall thickness, or plaque volume. None of the standard techniques can characterize the composition of an atherosclerotic plaque and therefore are incapable of identifying the high-risk plaques. High resolution, multicontrast, magnetic resonance imaging (MRI) can noninvasively image vulnerable plaques and characterize plaques in terms of their different components (ie, lipid, fibrous, calcium, or thrombus). Application of MRI opens up whole new areas for diagnosis, prevention, and treatment of atherosclerosis. PMID- 11473747 TI - Coronary artery disease progression assessed by electron-beam computed tomography. AB - The ability to observe changes in atherosclerotic plaque burden over time should provide an accurate measure of efficacy for different cardiovascular therapies. Electron-beam computed tomography (EBCT), by quantification of coronary artery calcification, is a noninvasive measure of atherosclerosis burden. This article summarizes data from abstracts and publications related to coronary artery calcium measurement and its use in progression studies. The issues related to interscan variability and reproducibility of this measure are detailed. The limitations of multidetector spiral computed tomography (high radiation dose and poor reproducibility) are also addressed. Several studies of progression using 2 scans, administered > or =1 year apart, demonstrate significant annual progression (22% to 52% per year). All studies demonstrate that therapy with cholesterol-lowering agents slows the atherosclerotic process, and that it may lead to regression of coronary calcium over time. There are 2 small prognostic studies that demonstrate that coronary events predominantly occur in those patients who exhibit significant progression of coronary artery calcium. Large multicenter trials are underway to evaluate the prognostic significance of coronary artery calcium progression. The progression of coronary artery atherosclerosis can be observed noninvasively by monitoring the progression of coronary calcification with EBCT. With annual progression rates of 22% to 52% and a median interscan variability of only 5% to 8%, this technology provides an opportunity to noninvasively monitor patients to assess the clinical efficacy of medical therapies in studies as short as 1 year. PMID- 11473748 TI - Nuclear cardiology and electron-beam computed tomography: competitive or complementary? AB - Electron-beam computed tomography (EBCT) and nuclear cardiology techniques are both valuable in the noninvasive assessment of patients with suspected coronary artery disease. The techniques, however, are different in the information they provide about the patient. EBCT provides anatomic information on coronary atherosclerosis, whereas myocardial perfusion single-photon emission computed tomography assesses the physiologic significance of coronary stenosis. Because of these differences, the techniques are highly complementary. In considering the complementary nature of these methods, it is important to clarify the issues being raised. An important question in the consideration of a patient with known or suspected coronary artery disease is, What is the risk in an individual patient of developing clinical coronary artery disease? The answer to this question will determine who needs aggressive medical management. A second question in a suspected coronary artery disease patient is, What is the risk of cardiac death? As will be discussed, this risk, in general, determines the need to consider coronary revascularization. In the former question, EBCT testing and clinical assessment alone is usually sufficient, and in some cases nuclear testing can be of additional value. In answering the second question, on the basis of currently available data, the EBCT and nuclear cardiology studies appear to be operating in a complementary fashion. PMID- 11473749 TI - Lipid disorders and plaque imaging. AB - Electron-beam tomography (EBT) plaque imaging has the potential to fundamentally change the practice of prevention, through application of the following principles: (1) Asymptomatic patients with significant calcified plaque should be considered to have a clinical coronary artery disease risk equivalent. (2) Measurement of nontraditional parameters should be incorporated into the evaluation of patients with calcified plaque. (3) Drug treatment should focus on the asymptomatic patient with subclinical atherosclerosis. (4) Change in plaque rather than change in lipid values should guide treatment. PMID- 11473750 TI - Clinical trials: surrogate endpoints or hard endpoints? AB - A man, aged 50, has about a 50% risk and a 50-year-old woman a 35% risk of having a myocardial infarction during their lifetime. The extent of atherosclerosis is the primary determinant of the risk of myocardial infarction. We will be unable to substantially reduce the lifetime risk of coronary artery disease without primary prevention of atherosclerosis. Noninvasive methods to measure subclinical atherosclerosis and its progression offer a unique opportunity to improve individual patient preventive strategies, based both on pharmacologic and nonpharmacologic therapies, as well as an opportunity to study and develop new drug therapies. The use of subclinical disease as a surrogate marker of atherosclerosis and the study of plaque characteristics will greatly enhance our understanding of the role of new risk factors for atherosclerosis, lipoprotein metabolism, and genetic-lifestyle interactions. PMID- 11473751 TI - From clinical trials to public health policy: the path from imaging to screening. PMID- 11473752 TI - Tomographic (plaque) imaging: state of the art. AB - Tomographic coronary artery plaque imaging is possible noninvasively using x-ray computed tomography (CT) and magnetic resonance imaging (MRI). The pathophysiology of coronary plaque disease is one of repeated inflammation and repair. Imaging of coronary artery calcium, a consequence of this process, is possible using CT, whereas MRI has the potential to examine the lipid and fibrous components of plaque acquisition and 3-dimensional slice registration. Quantitation of coronary artery calcium has been validated using electron-beam CT [EBCT], a unique device that images the entire heart in a single breathold from rapid [100 msec] tomographic scans done in synchrony with the heart cycle. Current mechanical CT devices require 300 to 500 msec per scan and acquire hundreds of tomographic images that then must be retrospectively separated to the required phase of the heart cycle. There are limited correlations with calcium scoring by EBCT versus mechanical CT and the later device has a limitation in situations of low plaque volume and necessitates increased radiation exposure to the patient. MRI has been shown to have the potential to define plaque composition ex vivo or in the aorta, but studies of the heart arteries are so far very limited. Widespread utilization of noninvasive plaque imaging requires that the studies be done consistently and reproducibly. The training of the interpreting physicians is of paramount concern. PMID- 11473753 TI - Noninvasive coronary angiography by magnetic resonance imaging, electron-beam computed tomography, and multislice computed tomography. AB - In recent years, several techniques for noninvasive imaging of the coronary artery lumen (noninvasive coronary angiography) have been developed. These techniques include magnetic resonance imaging, electron-beam computed tomography, and, most recently, multislice computed tomography. Each of these techniques has specific advantages and disadvantages. Currently, EBCT seems to permit the most robust coronary artery imaging. In the future, imaging modalities will have to be further improved and validated in order to define specific areas for potential clinical applications. PMID- 11473785 TI - Single molecule measurements of titin elasticity. AB - Titin, with a massive single chain of 3--4MDa and multiple modular motifs, spans the half-sarcomere of skeletal and cardiac muscles and serves important, multifaceted functions. In recent years, titin has become a favored subject of single molecule observations by atomic force microscopy (AFM) and laser optical trap (LOT). Here we review these single titin molecule extension studies with an emphasis on understanding their relevance to titin elasticity in muscle function. Some fundamental aspects of the methods for single titin molecule investigations, including the application of dynamic force, the elasticity models for filamentous titin motifs, the technical foundations and calibrations of AFM and LOT, and titin sample preparations are provided. A chronological review of major publications on recent single titin extension observations is presented. This is followed by summary evaluations of titin domain folding/unfolding results and of elastic properties of filamentous titin motifs. Implications of these single titin measurements for muscle physiology/pathology are discussed and forthcoming advances in single titin studies are anticipated. PMID- 11473786 TI - Analysis of single-molecule mechanical recordings: application to acto-myosin interactions. AB - Several laboratories have now developed methods to make single-molecule mechanical recordings from interacting pairs of biological molecules. The mechanical work done (product of force and distance) by a single biomolecular interaction is usually of the same order as thermal energy. Recordings made from non-processive, intermittently interacting, molecular motors such as acto-myosin therefore contain a large background of thermal noise. We have applied Page's test to analyse mechanical interactions between muscle myosin II's and F-actin recorded using an optical tweezers based single-molecule mechanical transducer. We compare Page's test with other variance-based methods and find it to be a robust method for analysing both simulated and real data sets. We discuss some of the problems associated with automatic detection of transient mechanical events in noisy data signals, and show that if the start and end points of individual events are known accurately then the events may be synchronised and combined to give more detailed information about different mechanical states. PMID- 11473787 TI - Structural and functional imaging with carbon nanotube AFM probes. AB - Atomic force microscopy (AFM) has great potential as a tool for structural biology, a field in which there is increasing demand to characterize larger and more complex biomolecular systems. However, the poorly characterized silicon and silicon nitride probe tips currently employed in AFM limit its biological applications. Carbon nanotubes represent ideal AFM tip materials due to their small diameter, high aspect ratio, large Young's modulus, mechanical robustness, well-defined structure, and unique chemical properties. Nanotube probes were first fabricated by manual assembly, but more recent methods based on chemical vapor deposition provide higher resolution probes and are geared towards mass production, including recent developments that enable quantitative preparation of individual single-walled carbon nanotube tips [J. Phys. Chem. B 105 (2001) 743]. The high-resolution imaging capabilities of these nanotube AFM probes have been demonstrated on gold nanoparticles and well-characterized biomolecules such as IgG and GroES. Using the nanotube probes, new biological structures have been investigated in the areas of amyloid-beta protein aggregation and chromatin remodeling, and new biotechnologies have been developed such as AFM-based haplotyping. In addition to measuring topography, chemically functionalized AFM probes can measure the spatial arrangement of chemical functional groups in a sample. However, standard silicon and silicon nitride tips, once functionalized, do not yield sufficient resolution to allow combined structural and functional imaging of biomolecules. The unique end-group chemistry of carbon nanotubes, which can be arbitrarily modified by established chemical methods, has been exploited for chemical force microscopy, allowing single-molecule measurements with well-defined functionalized tips. PMID- 11473789 TI - The golden age of rapid eye movement sleep discoveries. 1. Lucretius--1964. AB - Although there were several premonitory signs of a sleep stage with dreaming, it was only in 1953 that such a stage was identified with certainty. This paper analyses the observations and research related to this dreaming stage (rapid eye movement sleep) until 1964. During these 11 years of research, the main psychological and physiological characteristics of this sleep stage were first described. Where the few results or discussions were later questioned, today's current state of knowledge is briefly outlined. PMID- 11473790 TI - Alternative RNA splicing in the nervous system. AB - Tissue-specific alternative splicing profoundly effects animal physiology, development and disease, and this is nowhere more evident than in the nervous system. Alternative splicing is a versatile form of genetic control whereby a common pre-mRNA is processed into multiple mRNA isoforms differing in their precise combination of exon sequences. In the nervous system, thousands of alternatively spliced mRNAs are translated into their protein counterparts where specific isoforms play roles in learning and memory, neuronal cell recognition, neurotransmission, ion channel function, and receptor specificity. The essential nature of this process is underscored by the finding that its misregulation is a common characteristic of human disease. This review highlights the current views of the biological phenomenon of alternative splicing, and describes evidence for its intricate underlying biochemical mechanisms. The roles of RNA binding proteins and their tissue-specific properties are discussed. Why does alternative splicing occur in cosmic proportions in the nervous system? How does it affect integrated cellular functions? How are region-specific, cell-specific and developmental differences in splicing directed? How are the control mechanisms that operate in the nervous system distinct from those of other tissues? Although there are many unanswered questions, substantial progress has been made in showing that alternative splicing is of major importance in generating proteomic diversity, and in modulating protein activities in a temporal and spatial manner. The relevance of alternative splicing to diseases of the nervous system is also discussed. PMID- 11473791 TI - Rapidly exchanging Ca2+ stores in neurons: molecular, structural and functional properties. PMID- 11473792 TI - Aversive learning in patients with unilateral lesions of the amygdala and hippocampus. AB - The present study applied a visual half field paradigm with emotional facial expressions in patients with selective unilateral amygdalo-hippocampectomy (AHE) to elucidate the contributions of the left and right medial temporal lobe and amygdala to emotional learning. Electrodermal indicators of aversive learning were studied in 14 left AHE and 12 right AHE patients, as well as 13 controls matched in sex and age. In a differential conditioning paradigm with negative (CS+) and positive (CS-) facial expressions, CS+ were associated with an aversive vocalization (US, 95 dB, 3 s). During extinction, stimuli were presented laterally and preattentively using backward masking. Appropriate CS durations yielding preattentive presentation were individually determined prior to conditioning. In contrast to controls, both left and right AHE patients failed to show an autonomic conditioning effect following left visual field presentations of masked negative CS+ during extinction. AHE patients also showed no clear differential acquisition. Moreover, right AHE patients poorly recognised that negative valence was an affiliating dimension of the CS-US compound. PMID- 11473793 TI - Genetic analysis of autonomic reactivity to psychologically stressful situations. AB - We present the results of a behavioural genetics study on response profiles of autonomic measures (heart rate, blood pressure, and galvanic skin level), under ecologically valid, stressful conditions. Where response profiles of different physiological variables are the object of study, and when daily life stressors are taken into account (Turner and Hewitt, Annals of Behavioral Medicine, 14 (1992) 12-20), autonomic responsiveness to psychological stressors is thought to be an inherited trait. The participants were 100 female twin pairs, 57 monozygotic and 43 dizygotic twin pairs. Participants watched eight films with a stressful social content while autonomic measures were continuously recorded. Results show that the heritability coefficients of response profiles of autonomic measures are almost twice as high as that of single variables. The results further show that genes exert their influence on physiological behaviour not only directly, but also indirectly, by influencing the idiosyncratic relation between a person and his environment. PMID- 11473794 TI - Dominance and testosterone in women. AB - Fifty-two young women completed the Simple Adjective Test (a questionnaire designed to measure dominance) and at the same time provided 5 ml blood for testosterone assay. Higher dominance scores were associated with higher serum testosterone levels (t-test P<0.008). PMID- 11473795 TI - Habituation of male sexual arousal: effects of attentional focus. AB - Three experiments are reported demonstrating that levels of penile tumescence and subjective sexual arousal are greater when men employ participant-oriented rather than spectator-oriented attentional focus while viewing an erotic film segment. Under each instructional set, there was a reduction in sexual arousal during repeated erotic stimulation. As sexual arousal habituated, the men reported feeling less absorbed during erotic stimulation. When these associated changes in attentional focus (absorption) were partialled out through analysis of covariance, sexual arousal remained relatively stable over trials, suggesting that sexual arousal is less likely to habituate if attentional focus remains constant during repeated erotic stimulation. Further directions for studying associations between habituation of sexual arousal and cognitive processing are discussed. PMID- 11473796 TI - Differences in naming accuracy of odors presented to the left and right nostrils. AB - Two interlinked areas of debate within psychology are the existence of hemispheric specialization for olfactory processing and whether odors are processed primarily as perceptual codes or as a set of semantic features. This study compared accuracy in naming and judgments of familiarity by right handed subjects to common odors that were presented to the left or right nostrils. There was significantly better production of the correct name of the odor after left side presentation, than after right. Familiarity ratings were identical. PMID- 11473797 TI - Interhemispheric asymmetry of EEG alpha activity at rest and during the Wisconsin Card Sorting Test: relations with performance. AB - There are conflicting results regarding the functional asymmetry of the prefrontal cortex. Spectral power analysis of electroencephalographic (EEG) activity can provide important clues about the cortical mechanisms. In this study, interhemispheric EEG alpha power asymmetry of healthy individuals was investigated during the execution of the Wisconsin Card Sorting Test (WCST) and during rest. We analyzed alpha-1 (8.6-10.2 Hz) and alpha-2 (10.9-12.5 Hz) bands separately and found some evidence to indicate that lower and upper alpha bands reflect different cortical processes. On the other hand, greater alpha power during resting correlated with higher performance on the WCST. The lower left frontal alpha power during WCST correlated significantly with the higher WCST performance. However, greater bilateral parietal alpha power during WCST correlated with higher performance. Significant correlations between EEG activity and WCST performance were, in general, restricted to lower alpha power, both at rest and during the task. These findings are discussed with regard to attention processes reflected by lower alpha activity. PMID- 11473798 TI - Application of solvent microextraction to the analysis of amphetamines and phencyclidine in urine. AB - A fast and simple method to detect some commonly abused illicit drugs, amphetamine, methamphetamine, 3,4-methylendioxy-amphetamine (MDA), 3,4 methylendioxy-methamphetamine (MDMA), 3,4-methylendioxy-N-ethylamphetamine (MDEA) and phencyclidine (PCP) in urine using solvent microextraction (SME) combined with gas chromatography (GC) analysis has been developed. The extraction is conducted by suspending a 2 microl drop of chloroform in a 2 ml urine sample. Following 8 min of extraction, the organic solvent is withdrawn into the syringe and injected into a GC with a pulsed discharge helium ionization detector (PDHID). The effects of different extraction solvents and times, pH and sample preparation were studied. The optimized method was capable of detecting drugs in urine at concentrations below Substance Abuse and Mental Health Services Administration (SAMHSA) established cut-off values for preliminary testing. Good linearity and reproducibility of extraction were obtained. The limits of detection were 0.5 microg/ml for amphetamine, 0.1 microg/ml for methamphetamine and MDA, 0.05 microg/ml for MDMA, 0.025 microg/ml for MDEA and 0.015 microg/ml for PCP. Relative standard deviation (R.S.D.) values ranged between 5 and 20% for the studied drugs. PMID- 11473799 TI - Powder method for detecting latent fingerprints: a review. AB - The powder technique for detecting latent fingerprints involves the application of a finely divided formulation to the fingermark impression, generally with a glass-fibre or a camel hair brush. The powder gets mechanically adhered to the sweat residue defining the ridge pattern. The furrows which are devoid of the fingerprint residue, do not adhere the powder onto them. The final outcome is that the powder formulation sticks to the ridges, but is easily blown off the furrows. Since the powder is normally coloured, the ridge pattern becomes visible and the latent print is said to have developed. PMID- 11473800 TI - Validation and casework application of a Y chromosome specific STR multiplex. AB - A series of validation experiments was performed for a Y chromosome specific STR multiplex system following the suggestions made by the Technical Working Group DNA Analysis Methods (TWGDAM). The multiplex PCR products were detected on Perkin Elmer 373 and 377 automated sequencers using two labeling colors. No problems regarding the stability, robustness and sensitivity of the Y STR multiplex were observed. Mixture studies revealed a cut off rate similar to autosomal STRs for mixtures of male DNAs and no interference of any female admixture. The comparison of the Y STR results to the autosomal typing results for 56 nonprobative semen stains and swabs, showed a slightly higher success rate in detecting the semen donor's alleles for the Y STR multiplex. Two examples are shown to illustrate the usefulness of Y STR typing for DNA mixtures. In one case the Y STR results confirmed an isolated exclusion; in the other case, the interpretation of a mixture was clarified since the Y STR results proved the presence of DNA from at least two semen donors. Y STR typing is a valuable addition to the forensic DNA testing panel. PMID- 11473801 TI - Immunohistochemical quantification of pulmonary mast-cells and post-mortem blood dosages of tryptase and eosinophil cationic protein in 48 heroin-related deaths. AB - Recent studies suggest that many fatal heroin overdoses are caused by anaphylactoid reaction. In the present study we measured tryptase and eosinophil cationic protein in post-mortem blood of 48 deaths after heroin injection. We also investigated the presence and pulmonary distribution of mast-cells using specific immunohistochemical antibody for tryptase and morphometric evaluation in those cases of heroin-related deaths. The data were compared with 44 subjects who died following head trauma and to 32 cases of fatal anaphylactic shock. In the heroin-related death cases, the measurements of serum tryptase levels and eosinophil cationic protein dosages resulted in particularly elevated concentrations compared with the trauma cases. Nevertheless, the data that our study supplies by immunohistochemical techniques indicate that when mast-cells count in the lung was determined, no definite pattern was obtained between fatal heroin overdose cases and the control groups. Furthermore, the wide range of morphine concentrations found in post-mortem blood samples suggest that the term 'overdose' is relative and does not sufficiently characterize death associated with heroin addiction. Our study confirms that elevated concentrations of serum tryptase are associated with many heroin-related deaths. At this moment to attribute the cause of these deaths to 'heroin overdose' ignores the likely causal contribution of other possible systemic reactions to the mechanism of death. PMID- 11473802 TI - Driving under the influence of alcohol and/or drugs in the Netherlands 1995-1998 in view of the German and Belgian legislation. AB - This study presents the test results of blood and urine samples of impaired drivers in the Netherlands between January 1995 and December 1998. In this period, the blood alcohol concentrations of 11,458 samples have been determined and 1665 blood or urine samples have been analysed for drugs. The median alcohol concentration was between 1.7 and 1.8 mg/ml blood. In 80% of the 1665 analysed samples drugs were detected. At least 42% (702/1665) of the impaired drivers were poly-drug users, with cocaine present in the most frequent combinations. In the Netherlands, the procedure to prove driving under the influence is complex. This procedure can be made more efficient and more effective by embedding the analytical test results, needed to prosecute an impaired driver, in the law. In Belgium and Germany, such laws already are in force. If we would apply the qualifications of the new Belgian law on our analytical data, 67% of the impaired drivers included in this comparison could have been prosecuted without discussion in court. PMID- 11473803 TI - Sequence polymorphism in the coding region of mitochondrial genome encompassing position 8389-8865. AB - Analysis of the polymorphic sequences in mitochondrial DNA (mtDNA) has been widely applied to forensic tests and anthropology studies. However, these polymorphic data in human have thus far been derived from the displacement-loop and intergenic regions only. Here, we report the identification of clustered polymorphic sites in the mitochondria coding region encompassing position 8389 8865. The DNA sequences of 119 unrelated Chinese were determined by PCR amplification and direct sequencing. The results showed that heteroplasmy was found in five individuals, 39 sites were noted in this 477 bp region, and 41 haplotypes were identified. The probability of identity and allelic diversity were estimated as 0.1265 and 0.8809, respectively. The results suggest that sequence polymorphism from position 8389-8865 in human mtDNA can be used as a marker for identity investigation. PMID- 11473804 TI - Y-chromosomal STR haplotype in Toscany (central Italy). AB - Here we show the Y-haplotype database consisting in the loci DYS19, DYS388, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385, YCAII and DXYS156Y of 107 males living in Toscany (central Italy). PMID- 11473805 TI - Multiplex DNA typing of short tandem repeat loci on Y chromosome of Chinese population in Taiwan. AB - Allele frequencies and haplotypes for ten Y chromosome STRs loci, namely, DYS19, DYS385 I, DYS385 II, DYS388, DYS389 I, DYS389 II, DYS390, DYS391, DYS392 and DYS393 were obtained from a sample of 582 Chinese individuals in Taiwan. PMID- 11473806 TI - Population data of Y-chromosomal STRs in Lithuanian, Latvian and Estonian males. AB - Data of nine Y-chromosomal STRs, the so called "extended core set", were obtained from 152 unrelated males from Lithuania, 145 from Latvia and 133 from Estonia. The haplotype data reported here have been included into the Y-STR database maintained at the Institute of Legal Medicine, Humboldt-University, Berlin. PMID- 11473807 TI - Human Y-specific STR haplotypes in a Slovenian population sample. AB - The allele distribution of the systems DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385 and YCAII were investigated in a sample of 121 unrelated males from Slovenia PMID- 11473808 TI - Gene transfer into retinal ganglion cells by in vivo electroporation: a new approach. AB - We developed a new in vivo electroporation method to deliver genes into retinal ganglion cells (RGCs). Efficiency and degree of tissue damage were evaluated using green fluorescent protein (GFP) gene and TUNEL. Soon after the intravitreous injection of the GFP gene, electroporation (five electric pulses of 99 ms duration each and 12V/cm delivered twice 5 min apart) was carried out on the adult rat eyeball with the aid of tweezer-type disc electrodes attached to corneal (cathode) and scleral (anode) surfaces. GFP expression, exhibiting a maximum on day 7, was detectable for up to 21 days. DiI retrograde labeling of RGCs showed that 41.5% of the total ganglion cells in the electroinjected area were GFP-positive. Therefore, this new method may be a useful tool for the delivery of genes into RGCs. PMID- 11473809 TI - Rapana thomasiana hemocyanin (RtH): dissociation and reassociation behavior of two isoforms, RtH1 and RtH2. AB - Rapana thomasiana hemocyanin (RtH) is a mixture of two hemocyanin isoforms, termed RtH1 and RtH2. The two subunit types, purified by ion exchange chromatography, were used for macromolecular reassociation studies. In vitro reassociation was achieved with Tris-saline stabilizing buffer at pH 7.4, containing 100mM calcium and magnesium chloride at 4 degrees C. The relatively slow progress of reassociation was monitored, and the different oligomeric forms of RtH1 and RtH2 were studied by transmission electron microscopy, using samples negatively stained with 1% (w/v) uranyl acetate or 5% (w/v) ammonium molybdate containing 1% (w/v) trehalose at pH 7.0. The two subunits reassociate to produce characteristic didecamers, oligomeric and polymeric forms depending on the dissociated material and the reassociation conditions (i.e. divalent ion concentration, duration). In contrast to the didecamers of the freshly isolated RtH preparations, RtH1 and RtH2 show after 2 weeks' reassociation a clear tendency to generate multidecameric structures. The behavior of the native RtH1 and RtH2 during reassociation in the presence of 100mM calcium and magnesium chloride corresponds to the reported common oligomerization characteristics of KLH1/HtH1 and KLH2/HtH2, respectively. It is important to note that during the reassociation of the RtH isoforms: (I) no smaller diameter tubular polymers (ca. 25-27nm) were formed from the subunits as well as from the decamers; (II) multidecamers with one or more 'nucleating' didecamers were detected in addition to the multidecamers, composed of didecamers with associated decamers at one or both ends. PMID- 11473810 TI - Correction of aberration for a high-resolution electron hologram by means of the amplitude contrast criterion of image wave. AB - In order to further improve the resolution for a high-resolution electron hologram, the aberration working on the hologram must be corrected. Since it is rather difficult to precisely control aberration coefficients in the experimental stage, we proposed an amplitude contrast D criterion of imaging wave to determine the working aberration from the hologram itself. In the determination or correction of the aberration, we assume a symmetrical aberration function is parameterized only by a spherical aberration coefficient and a defocus value. First, D is calculated from a holographically reconstructed imaging wave of YBa(2)Cu(3)O(7-x) for each combination of these parameters. The working aberration on the imaging wave is determined from the combination of the parameters by noting the maximum or minimum D of the imaging wave at some specifically chosen thickness regions. The theoretical validity for the D criterion is then proved with three-beam dynamical diffraction formula. Finally, the 'experimental' examination for the D criterion is successfully performed on the reconstructed image wave for the Sigma=9 interface structure of a wedge shaped silicon sample. PMID- 11473811 TI - Selective photoreceptor damage in four species of insects induced by experimental exposures to UV-irradiation. AB - Damage to photoreceptive cells of insect compound eyes exposed to abnormally high doses of UV-radiation of 350nm peak wavelength manifests itself in at least two different ways. In the butterflies Papilio xuthus and Pieris napi from Japan and northern Finland, respectively, only the cell bodies of retinula cells 1 and 2, (identified as short wavelength receptors), but not their corresponding rhabdomeres, exhibit damage with apoptotic features. In the eye of UV-irradiated adult crickets, however, cell bodies and cytoplasm remain normal, while the rhabdomeres of cells 7 and 8 exhibit signs of severe membrane disruptions. No signs of damage whatsoever occurred in the eyes of northern Finnish bumblebees exposed to UV. It is suggested that metabolic shortfalls in the UV-sensitive cells of the butterfly eyes result in cellular shut-down, but that in the cricket receptors UV-induced changes of the membrane lipids dominate, leading to membrane instability without concomittant cell death. The strong resistance of the bumblebee eye to UV-induced damage requires further investigation, but since preconditioning to light can reduce photic damage in the rat eye, the 24h daylight experienced by northern Finnish bumblebees during the summer season could be involved. PMID- 11473812 TI - Localization of transcripts corresponding to the major allergen from olive pollen (Ole e I) by electron microscopic non-radioactive in situ RT-PCR. AB - In situ reverse transcription-PCR of mRNAs corresponding to the olive major allergen (Ole e I) has been tested at the ultrastructural level in mature olive pollen. The transcripts were present in the cytoplasm of both the vegetative and the generative cells, frequently associated to ribosomes in the endoplasmic reticulum. No labeling was detected in the pollen wall, nor in vacuoles, lipid bodies, plastids or mitochondria. Localization of the major olive allergen at ultrastructural level showed the protein present mainly in the lumen of the endoplasmic reticulum vesicles or pockets scattered in the cytoplasm, and in the outer region of the pollen exine. The results confirm the rough endoplasmic reticulum as the cell system involved in both the synthesis and storage of this protein. This is the first report of in situ RT-PCR on plant material at the ultrastructural level. The method described for mRNA amplification and detection is confirmed as a valuable tool for studying gene expression in plant material. PMID- 11473813 TI - A tilting procedure to enhance compositional contrast and reduce residual diffraction contrast in energy-filtered TEM imaging of planar interfaces. AB - This paper systematically demonstrates that energy-filtered transmission electron microscope (EFTEM) images of a planar interface between two single crystals have increased compositional contrast and decreased residual diffraction contrast when the sample is oriented so that the electron beam is parallel to the interface, but not directly on a zone axis. This off-axis orientation reduces diffraction contrast in the unfiltered (and zero-loss) image, which in turn, reduces residual diffraction contrast in single energy-filtered TEM (EFTEM) images, thickness maps, jump-ratio images, and elemental maps. Most importantly, this procedure produces EFTEM images that are more directly interpretable and, in most cases, possess superior spatial resolution compared to EFTEM images acquired directly on a zone axis. PMID- 11473814 TI - Ferritin in iron containing granules from the fat body of the honeybees Apis mellifera and Scaptotrigona postica. AB - It is already known that the behaviour of the honeybee Apis mellifera is influenced by the Earth's magnetic field. Recently it has been proposed that iron rich granules found inside the fat body cells of this honeybee had small magnetite crystals that were responsible for this behaviour. In the present work, we studied the iron containing granules from queens of two species of honeybees (A. mellifera and Scaptotrigona postica) by electron microscopy methods in order to clarify this point. The granules were found inside rough endoplasmic reticulum cisternae. Energy dispersive X-ray analysis of granules from A. mellifera showed the presence of iron, phosphorus and calcium. The same analysis performed on the granules of S. postica also indicated the presence of these elements along with the additional element magnesium. The granules of A. mellifera were composed of apoferritin-like particles in the periphery while in the core, clusters of organised particles resembling holoferritin were seen. The larger and more mineralised granules of S. postica presented structures resembling ferritin cores in the periphery, and smaller electron dense particles inside the bulk. Electron spectroscopic images of the granules from A. mellifera showed that iron, oxygen and phosphorus were co-localised in the ferritin-like deposits. These results indicate that the iron-rich granules of these honeybees are formed by accumulation of ferritin and its degraded forms together with elements present inside the rough endoplasmic reticulum, such as phosphorus, calcium and magnesium. It is suggested that the high level of phosphate in the milieu would prevent the crystallisation of iron oxides in these structures, making very unlikely their participation in magnetoreception mechanisms. They are most probably involved in iron homeostasis. PMID- 11473815 TI - Use of light and scanning electron microscopy to examine colonisation of barley rhizosphere by the nematophagous fungus Verticillium chlamydosporium. AB - Barley roots were readily colonised by the nematophagous fungus Verticillium chlamydosporium. Light microscopy (LM) but also low temperature scanning electron microscopy (LTSEM) revealed details of the colonisation process. Hyphae were found on the rhizoplane often with dictyochlamydospores. Hyphae of V. chlamydosporium penetrated epidermal cells, often by means of appressoria. A hyphal network was formed in epidermal and cortical cells. Likewise, hyphal coils were found within root cells next to transverse cell walls. Cortical cells were the limits of fungal colonisation, since no hyphae were seen in the vascular cylinder. Modifications of root cell contents (phenolic droplets and callose appositions) were common three weeks after inoculation with V. chlamydosporium. These features may indicate induction of plant defence reactions in late stages of root colonisation by the fungus. Both LTSEM and LM have proved extremely useful to describe root colonisation by the fungus. The results found may have implications in the mode action of nematophagous fungi against plant parasitic nematodes. PMID- 11473816 TI - A new type of scanning electron microscope using the coaxial backscattered electrons. AB - A new coaxial detection system for backscattered electrons in SEM is described. This coaxial detection system allows us to collect only the backscattered electrons that have lost a small percentage of the primary energy, emerging from the sample surface with a take-off angle defined by the objective lens. This new configuration reinforces the atomic-number contrast and suppresses effectively the topographic contrast. The simulation and experimental results confirm these expectations: this new type of SEM is very suitable for observing differences in atomic number. Moreover, by associating the obtained image with a conventional secondary electron image, we build a third (color) image that allows us to give finally at the same time, in a single image, both of the chemical and topographic information. PMID- 11473817 TI - Caveolae and caveolin isoforms in rat peritoneal macrophages. AB - Caveolea are special (highly hydrophobic) plasma membrane invaginations with a diameter of 50-100 nm. Their characteristic features are the flask- or omega shape and the lack of basket-like coat composed of clathrin. Caveolin-an integral membrane protein-is the principal component of caveolae membranes in vivo. Multiple forms of caveolin have been identified: caveolin-1alpha, caveolin-1beta, caveolin-2 and caveolin-3. They differ in their specific properties and tissue distribution. In this paper we summarize the morphological and biochemical data providing strong evidence about the existence and function of caveolae in rat peritoneal macrophages. When studied electron microscopically, the surface of both resident and elicited macrophages exhibited omega- or flask-shaped plasma membrane invaginations. There was a significant difference, however, in the number of these profiles: whereas in resident cells only a small amount of them was found on the cell surface, in elicited cells they were abundantly present on the plasma membrane. Using an antibody against the VIP21/caveolin-1 isoform we showed that these plasma membrane pits were indeed caveolae. The number and the appearance of caveolae were found to be in close correlation with the functional activity of these phagocytotic cells, indicating that the formation of caveolae is a highly regulated process. Using Western blot analysis two different proteins ( approximately 29 and approximately 20 kDa)-both labelled with anti-caveolin antibodies-were identified in resident and elicited macrophages that have been isolated from rat peritoneal cavity. The approximately 20 kDa protein was labelled specifically only by anti-VIP21/caveolin-1, while the approximately 29 kDa protein was labelled by both anti-VIP21/caveolin-1 and anti-caveolin-2 antibodies. The presence of the approximately 29 kDa protein was highly characteristic of resident cells, and only a small amount of approximately 20 kDa protein was detected in these cells. Elicitation has resulted in a significant increase in the amount of approximately 20 kDa protein labeled only with anit VIP21/caveolin-1. Our morphological (confocal and electron microscopical) studies have shown that in resident cells caveolin was present in the cytoplasm, in smaller vesicles and multivesicular bodies around the Golgi area. Only a very small amount of caveolae was found on the cell surface of these cells. In elicited macrophages, caveolae (labelled with anti-VIP21/caveolin-1 antibody) appeared in large numbers on the cell surface, but caveolin detected by anti caveolin-2 was also found in small vesicles and multivesicular bodies. These data support the idea that the expression of the approximately 29 kDa (caveolin related) protein is insufficient for caveolae formation in resident cells, it can function as a modified, macrophage-specific caveolin-2 isoform. Our results strongly suggest that caveolin-1 plays a crucial role in the formation of caveolae: it is the amount of caveolin-1 that regulates the appearance of caveolae on the plasma membrane. Studying the endocytotic processes of resident and elicited macrophages we have found that elicited macrophages bound and internalized significantly larger amounts of fluid phase marker (HRP) and immune complex (peroxidase-antiperoxidase-PAP) than resident cells. Serial section analysis, double labelled immunocytochemistry, and filipin treatment were used to demonstrate that caveolae can pinch off from the plasma membrane and can take part in endocytotic processes as alternative carriers in elicited macrophages. PMID- 11473818 TI - Specific staining of tissue components with metal-hematoxylin complexes. AB - The older metal-hematoxylin stains stain a broad spectrum of tissue components. Several recently introduced metal-hematoxylin stains are highly selective. This selectivity is usually bought at the price of severe limitations on the choice of fixative. A very dilute (2 x 10(-4)M) aluminum hematoxylin is selective for nucleic acids in tissues fixed in organic solvents alone. Vanadate hematoxylin is selective for basic proteins in tissues fixed in formaldehyde or mercuric salts. Bismuth hematoxylin is selective for arginine residues and thus for histones and myelin basic protein in tissues fixed in strong acids (Bouin's fluid or SUSA fluid). Zirconyl hematoxylin is selective for acidic mucins. Zirconyl hematoxylin does not restrict the choice of fixative. PMID- 11473819 TI - Cross-section preparation for transmission electron microscopy of phases and interfaces in C/BN heterostructures. AB - A technique is described for the preparation of transmission electron microscopy cross-sectional samples of pyrolytical carbon layers deposited on polycrystalline boron nitride substrates. To solve the problem of different abrasion rates of C and BN a filler material, Si wafers, has been bonded to both sides of the pre thinned BN substrate. Correspondence between color and thickness of Si wafers facilitates controlled sample thickness reduction during dimpling. The samples prepared by this technique even without ion milling are thin enough for HRTEM studies. PMID- 11473821 TI - Lymphocyte-mediated immunosurveillance of epithelial cancers? AB - The validity and importance of the immunosurveillance of tumors have been debated ever since Burnet and Thomas outlined several key concepts that underpin our understanding of tumor immunity. With the development of gene-targeting technology, it has been shown that interferon gamma (IFN-gamma) and perforin--key effector molecules of both the innate and adaptive immune systems--contribute to the prevention of of tumors in mice. These findings have rejuvenated interest in the immunosurveillance of tumors, and recent data have provided evidence that the immune system influences the development of some spontaneous epithelial malignancies. PMID- 11473822 TI - Stressed bacteria and TB vaccines. PMID- 11473823 TI - Crohn's disease gene is given the NOD. PMID- 11473824 TI - Another bell tolls for Toll-like receptors. PMID- 11473825 TI - A tale of T-cell tubulin. PMID- 11473827 TI - Vaccine safety concerns. PMID- 11473826 TI - Blocking lymphotoxin: a potential therapy for diabetes? PMID- 11473829 TI - Irish say no to Nice but yes to immunology. PMID- 11473828 TI - Toll-like receptors and the danger hypothesis. PMID- 11473830 TI - Antinuclear autoantibodies as potential antineoplastic agents. AB - The immune system confines neoplasia at various stages of tumor development. Whereas the role of cellular immunity has been investigated widely and utilized in the clinic, the importance of humoral immunity in this process has begun to emerge only in recent years. Circulating antinuclear autoantibodies (ANAs) typically found in autoimmune conditions, have also been detected in cancer patients and in healthy elderly individuals. The pathogenic role of ANAs in autoimmunity is well studied; however, little research has been carried out to elucidate the functions of ANAs in cancer patients. Experimental data favoring the antitumor activity of ANAs might support the clinical testing of monoclonal ANAs as a cancer therapy, if confirmed by further experiments. PMID- 11473831 TI - An etiopathogenic role for the type I IFN system in SLE. AB - The type I interferon (IFN) system plays a pivotal role in the etiopathogenesis of systemic lupus erythematosus (SLE). The initial appearance of autoantibody producing B cells can be precipitated by infection-induced type I IFNs, but the further, significant generation of autoimmune T and B cells is caused by the prolonged production of IFN-alpha, which is maintained by a vicious circle mechanism. This involves the activation of immature dendritic cells, known as natural IFN-producing cells, by continuously formed endogenous IFN-alpha inducers. These IFN-alpha inducers consist of complexes of autoantibodies with nucleic-acid-containing autoantigens derived from apoptotic cells. PMID- 11473832 TI - Neighborhood politics: the immunoregulatory function of organ-resident liver endothelial cells. AB - The liver is known for its ability to induce antigen (Ag)-specific immune tolerance. Among the different cell populations involved in the induction of hepatic tolerance, the liver sinusoidal endothelial cells (LSECs) are particularly important because they are highly efficient at presenting soluble Ags to CD4(+) and CD8(+) T cells. The crosspresentation of soluble Ags to CD8(+) T cells was believed previously to be restricted to professional Ag-presenting cells (APCs) such as dendritic cells (DCs). However, in contrast to DCs, crosspresentation by LSECs can induce Ag-specific immune tolerance. It is proposed that these organ-resident APCs act as sessile hepatic APCs that control the immune responses to soluble blood-borne Ags, in concert with APCs in lymphatic tissue. PMID- 11473833 TI - Designer dendritic cells for tolerance induction: guided not misguided missiles. AB - Dendritic cells (DCs) are professional antigen-presenting cells (APCs) that play crucial roles as initiators and modulators of adaptive immune responses. Although DC-based vaccines have been utilized successfully to generate cytolytic T-cell activity against tumor antigens (Ags), evidence has accumulated that DCs also have potent capabilities to tolerize T cells in an Ag-specific manner. DCs cultured in the laboratory can suppress auto- or alloimmunity. Current and prospective strategies to promote this inherent tolerogenic potential of DCs might prove to be important for the therapy of transplant rejection and autoimmune diseases. PMID- 11473834 TI - Post-translational protein modifications in antigen recognition and autoimmunity. AB - It is estimated that 50-90% of the proteins in the human body are post translationally modified. In the proper context, these modifications are necessary for the biological functions of a vast array of proteins and the effector functions of the cells in which they reside. However, it is now clear that some post-translational modifications can create new self antigens (Ags) or even mask Ags normally recognized by the immune system. In either case, they profoundly affect the recognition of Ag by bone marrow-derived cells, as well as their effector functions. How do post-translational protein modifications affect the processing of foreign and self Ags and what is their role in the origin of autoimmune responses? PMID- 11473835 TI - Th1- and Th2-cell commitment during infectious disease: asymmetry in divergent pathways. AB - The development of T helper 1 (Th1) versus Th2 cells is a major branch point in the immune response. It is an important determinant of whether the response to an infectious pathogen will lead to protection of the host or dissemination of the disease. Recent studies have suggested that this process is governed by distinct sets of signals provided by dendritic cells upon interactions with specific infectious agents. A model is proposed that links together the pathogen, the innate response and Th-cell polarization. PMID- 11473836 TI - DC-SIGN and LFA-1: a battle for ligand. AB - The intercellular adhesion molecules (ICAMs) play a prominent role in regulating the migration and activation of both dendritic cells (DCs) and T lymphocytes in the immune system. Recent observations have demonstrated that both leukocyte function-associated molecule 1 (LFA-1) and DC-specific ICAM-grabbing nonintegrin (DC-SIGN), two structurally unrelated adhesion receptors, regulate the function of leukocytes and DCs by binding to the same ICAMs. Here, we focus on the structure-function relationships of DC-SIGN and LFA-1 to obtain an insight into their role in the migration and activation of DCs and T cells in the control of immunity. PMID- 11473837 TI - Routine laboratory testing in the elderly: is it indicated? PMID- 11473838 TI - Cytokines during ventilator-induced lung injury: a word of caution. PMID- 11473839 TI - Thoracic epidural bupivacaine attenuates supraventricular tachyarrhythmias after pulmonary resection. AB - Supraventricular tachyarrhythmias after pulmonary surgery are well described. Some investigators suggest that tachyarrhythmias after thoracic operations may result from the relative sympathotonic status produced by injury to the cardiac parasympathetic nerves. We examined whether postoperative thoracic sympathetic blockade by thoracic epidural bupivacaine might reduce the tachyarrhythmias after pulmonary resection. Fifty patients with lung cancer were randomized to receive epidural bupivacaine (Group B) or epidural morphine (Group M). Patients in Group B were given 6 to 10 mL of 0.25% bupivacaine epidurally, followed by epidural infusion at 3 to 5 mL/h for 3 days, and patients in Group M were given 2 to 3 mg morphine epidurally, followed by morphine infusion at a rate of 0.2 mg/h. Tachyarrhythmias were diagnosed by using the continuous heart rate trend and arrhythmia trend with a central monitoring system. Postoperative analgesia was not statistically different between groups. However, the incidence of postoperative tachyarrhythmias in Group B was significantly less than in Group M (1 of 23 vs 7 of 25, P = 0.0497, Fisher's exact test). The continuous infusion of thoracic epidural bupivacaine can reduce supraventricular tachyarrhythmias compared with epidural morphine infusion, presumably because of attenuation of the sympathotonic status after pulmonary resection. IMPLICATIONS: We examined whether postoperative thoracic sympathetic blockade by thoracic epidural bupivacaine after pulmonary resection might reduce the tachyarrhythmias that may result from the relative sympathotonic status produced by injury to the cardiac parasympathetic nerves. The continuous infusion of thoracic epidural bupivacaine was shown to reduce supraventricular tachyarrhythmias. PMID- 11473840 TI - Ipsilateral shoulder pain after thoracotomy with epidural analgesia: the influence of phrenic nerve infiltration with lidocaine. AB - Patients receiving effective thoracic epidural analgesia for postthoracotomy pain may still complain of severe ipsilateral shoulder pain. The etiology of this pain is unclear. In this randomized, double-blinded, placebo-controlled study, we investigated the effect of phrenic nerve infiltration with lidocaine or saline on postoperative shoulder pain in 48 patients. After completion of a lung resection, patients received either 10 mL of 1% lidocaine or 10 mL of 0.9% saline infiltrated into the periphrenic fat pad at the level of the diaphragm. Shoulder pain was experienced by 33% of patients receiving lidocaine, compared with 85% of patients receiving saline (P < 0.008). Overall pain scores were lower with lidocaine (P < 0.05). PaCO(2) values were not significantly higher with lidocaine in the first 2 h. We conclude that pain transmitted via the phrenic nerve and referred to the shoulder is the most likely explanation for the ipsilateral shoulder pain experienced by patients receiving epidural analgesia for postthoracotomy pain. IMPLICATIONS: Ipsilateral shoulder pain after thoracotomy is common and may be severe, even in the presence of a functioning thoracic epidural. We have shown that infiltration of the phrenic nerve with local anesthetic significantly and safely reduces this shoulder pain, potentially allowing the ideal goal of a pain-free thoracotomy. PMID- 11473841 TI - Late preconditioning is blocked by racemic ketamine, but not by S(+)-ketamine. AB - Racemic ketamine blocks K(ATP) channels in isolated cells and abolishes short term cardioprotection against prolonged ischemia. We investigated the effects of racemic ketamine and S(+)-ketamine on ischemic late preconditioning (LPC) in the rabbit heart in vivo. A coronary occluder was chronically implanted in 36 rabbits. After recovery, the rabbits divided into four groups (each n = 9). LPC was induced in conscious rabbits by a 5-min coronary occlusion. Twenty-four hours later, the animals were instrumented for measurement of left ventricular systolic pressure (LVSP, tip manometer), cardiac output (CO, ultrasonic flowprobe) and myocardial infarct size (triphenyltetrazolium staining). All rabbits were then subjected to 30-min coronary occlusion and 2 h reperfusion. Controls underwent the ischemia-reperfusion program without LPC. To test whether racemic ketamine or S(+)-ketamine blocks the cardioprotection induced by LPC, the drugs (10 mg/kg) were given 10 min before the 30-min ischemia. Hemodynamic values were not significantly different between groups during the experiments (baseline: LVSP, 94 +/- 3 mm Hg [mean +/- SEM] and CO, 243 +/- 9 mL/min; coronary occlusion: LVSP, 93% +/- 4% of baseline and CO, 84% +/- 4%; after 2 h of reperfusion: LVSP, 85% +/ 4% and CO, 83% +/- 4%). LPC reduced infarct size from 44% +/- 3% of the area at risk in controls to 22% +/- 3% (P = 0.002). Administration of racemic ketamine abolished the cardioprotective effects of LPC (44 +/- 4%, P = 0.002). S(+) ketamine did not affect the infarct size reduction induced by LPC (26 +/- 6%, P = 0.88). IMPLICATIONS: Racemic ketamine, but not S(+)-ketamine, blocks the cardioprotection induced by ischemic late preconditioning in rabbit hearts in vivo. Thus, the influence of ketamine on ischemic late preconditioning is most likely enantiomer specific, and the use of S(+)-ketamine may be preferable in patients with coronary artery disease. PMID- 11473842 TI - Clonidine-induced vasoconstriction in awake volunteers. AB - We evaluated the vasomotor effects of clonidine in awake subjects with an intact central cardiovascular regulatory system. To determine the lower limit of the vasoconstrictive effect of clonidine in awake volunteers, we blocked sympathetic innervation to the left arm by anesthetizing the brachial plexus. We then measured arterial blood pressure and vasoconstriction via finger volume plethysmography measuring infrared light transmitted through a fingertip (LTF). LTF values obtained from the left arm were compared with those from the neurally intact right arm during four progressively increasing IV doses of clonidine, targeting plasma clonidine concentrations of 0.3, 0.45, 0.68, and 1.0 ng/mL. Clonidine decreased systolic blood pressure (P < 0.004) from 135 +/- 8 mm Hg to 115 +/- 8 mm Hg and heart rate (P = 0.0017) from 68 +/- 7 mm Hg to 61 +/- 10 mm Hg. Clonidine decreased LTF by -12% +/- 11% (P < 0.0001) less than preinfusion values at the 0.68 ng/mL target concentration in the right hand. In contrast, in the left hand, clonidine increased LTF significantly more than (P < 0.0001) preinfusion values at all target concentrations, with a maximal increase of 30% +/- 7%. We conclude that IV clonidine, at doses that decrease arterial blood pressure, causes arterial vasoconstriction in awake subjects. IMPLICATIONS: IV clonidine, at doses that decrease blood pressure, causes arterial vasoconstriction in awake subjects. These data suggest that an alpha-2 agonist with a high alpha-2a/alpha-2b selectivity should provide more profound sedative and analgesic effects with less undesirable vasoconstrictive effects. PMID- 11473843 TI - Arterial and venous Thrombelastograph variables differ during cardiac surgery. AB - The Thrombelastograph (TEG; Haemoscope Corp., Skokie, IL) coagulation analyzer is an effective point-of-care monitor for routine clinical practice and clinical research. Prior investigators have used either arterial or venous samples of blood for TEG measurements. We conducted this prospective cohort study to determine potential differences in TEG variables between arterial and venous blood samples. Arterial and venous samples were drawn from 40 cardiac surgical patients, yielding 134 pairs for comparison. Twenty-nine comparisons (control) were between arterial and arterial samples and were not significantly different. For the arterial and venous comparisons (n = 105), mean (+/-sd) arterial and venous values were the following: reaction time, 10 +/- 2 mm vs 13 +/- 4 mm, P = 0.004; maximum amplitude, 59 +/- 9 mm vs 49 +/- 12 mm, P < 0.001; alpha angle, 61 +/- 10 degrees vs 51 +/- 14 degrees, P < 0.001; K, 5 +/- 2 mm vs 8 +/- 4 mm, P = 0.007; and lysis, 2.5 +/- 1.7 vs 2.5 +/- 2.0 (not significant), arterial versus venous, respectively. Arterial blood samples demonstrated TEG values reflecting stronger (larger maximum amplitude) and faster (shorter reaction time and K value, wider alpha angle) clot formation. The results suggest that users of TEG coagulation analyzers should be consistent with the site of blood sampling given the potential differences obtained. IMPLICATIONS: Thrombelastograph (TEG) values obtained from venous blood samples differ from values obtained from arterial blood samples. When the TEG coagulation analyzer is used for clinical purposes, it is important to be consistent in the blood collection site. PMID- 11473844 TI - Cibenzoline has an inhibitory effect on vasorelaxation mediated by adenosine triphosphate-sensitive K(+) channels in the rat carotid artery. AB - Studies in cardiac myocytes have shown that cibenzoline reduces adenosine triphosphate (ATP)-sensitive K(+) currents, suggesting that this class Ia antiarrhythmic drug may modify the activity of ATP-sensitive K(+) channels in these preparations. The effects of class Ia antiarrhythmic drugs on vasodilation mediated by ion channels have not been studied. Therefore, we designed this study to examine whether cibenzoline may produce changes in vasorelaxation in response to a selective ATP-sensitive K(+) channel opener, levcromakalim, in the isolated rat carotid artery. Rings of rat carotid arteries without endothelium were suspended for isometric force recording. Concentration-response curves were obtained in a cumulative fashion. During submaximal contraction to phenylephrine (3 x 10(-7) M), vasorelaxation in response to levcromakalim (10(-8) to 10(-5) M) or 1-hydroxy-2-oxo-3-(N-methyl-3-aminopropyl)-3-methyl-1-triazene (NOC-7; 10(-10) to 10(-5) M) was obtained. During contraction to phenylephrine, levcromakalim induced concentration-dependent vasorelaxation. A selective ATP-sensitive K(+) channel antagonist, glibenclamide (5 x 10(-6) M), completely abolished vasorelaxation in response to levcromakalim, whereas a selective Ca(2+)-dependent K(+) channel antagonist, iberiotoxin (5 x 10(-8) M), did not affect the relaxation. Cibenzoline (10(-6) to 10(-5) M) significantly reduced vasorelaxation to levcromakalim in a concentration-dependent fashion. In contrast, cibenzoline (10(-5) M) did not alter vasorelaxation to a nitric oxide donor, NOC-7. These results suggest that from the clinically relevant concentrations, a novel class Ia antiarrhythmic drug, cibenzoline, impairs carotid vasodilation mediated by ATP sensitive K(+) channels. IMPLICATIONS: In isolated rat carotid artery, cibenzoline (10(-6) to 10(-5) M) reduced vasorelaxation to levcromakalim in a concentration-dependent fashion. These results suggest that from the clinically relevant concentrations, a novel class Ia antiarrhythmic drug, cibenzoline, impairs carotid vasodilation mediated by adenosine triphosphate-sensitive K(+) channels. PMID- 11473845 TI - An effective treatment of severe intractable bleeding after valve repair by one single dose of activated recombinant factor VII. AB - IMPLICATIONS: The successful treatment with recombinant factor VIIa of a patient experiencing intractable bleeding after cardiac surgery is described. PMID- 11473846 TI - Anterior interosseous nerve palsy after cardiopulmonary resuscitation in a resuscitator with undiagnosed muscle anomaly. AB - IMPLICATIONS: We present a case of nerve palsy after cardiopulmonary resuscitation in a resuscitator with undiagnosed muscle anomaly. Effort-related nerve palsy may occur after prolonged performance of CPR. PMID- 11473847 TI - Respiratory failure after pneumonectomy in a patient with unknown hyperlipidemia. AB - IMPLICATIONS: We report the case of a patient who had increased lipids in his blood and who complained of dyspnea the first postoperative day after resection of his left lung. As the blood lipids were decreased, his respiration was improved. We conclude that when respiration deteriorates postoperatively, increased blood lipids should be considered as a cause. PMID- 11473848 TI - Lord or vassal? Academic anesthesiology finances in 2000. AB - This article examines recent trends in the management of academic physician practice groups, and in particular the allocation of revenues and expenses to anesthesiology departments. The history of academic group practice is traced, beginning with the "corporate model," in which each department functioned in financial independence from the others. This evolved gradually into the "feudal system," in which departments were ostensibly independent, but paid variable and often large "assessments" to the central group. The final stage in this evolution is the "big bag," in which all clinical revenue is pooled by the central practice group, and then distributed by the group to departments or individuals according to some compensation plan formula. The advantages and disadvantages of each of these systems are discussed as they apply to anesthesiology departments. A productivity-based compensation plan formula under the big bag system is calculated for a typical anesthesiology department. This calculation shows that if the compensation formula is truly based on measured clinical productivity, anesthesiology departments may actually fare better under the big bag than under the feudal system. Finally, options for survival in the academic practice groups of the future are discussed. IMPLICATIONS: The history, current status, and trends of finances in academic anesthesiology departments are reviewed. Knowledge of these issues will help departments develop funds allocation methods to ensure that they receive an appropriate share of their faculty practice group's clinical income. PMID- 11473849 TI - The prevalence and predictive value of abnormal preoperative laboratory tests in elderly surgical patients. AB - Because data to determine which preoperative laboratory tests are important in elderly surgical patients are limited, we performed a prospective cohort study to evaluate the prevalence and predictive value of abnormal preoperative laboratory tests in consecutive patients > or =70 yr old who were undergoing noncardiac surgery. Patients presenting for surgery requiring only local anesthesia or monitored anesthesia care were excluded. Preoperative risk factors and laboratory test results were measured and evaluated for their association with the occurrence of predefined in-hospital postoperative adverse outcomes. In 544 patients, the prevalence of preoperative electrolytes and platelet count abnormalities (<115 x10(9)/L) was small (0.5%-5%), and abnormal creatinine (>1.5 mg/dL), hemoglobin (<10 g/dL), and glucose (>200 mg/dL) values were 12%, 10%, and 7%, respectively. Univariate predictors for adverse outcome of abnormal sodium and creatinine were not as predictive as ASA classification and surgical risk. By multivariate logistic regression, only ASA classification (>II) (odds ratio [OR], 2.55; 95% confidence interval [CI], 1.56-4.19; P < 0.001) and surgical risk (OR, 3.48; 95% CI, 2.31-5.23; P < 0.001) were significant independent predictors of postoperative adverse outcomes. The prevalence of abnormal preoperative electrolyte values and thrombocytopenia was small and had low predictive values. Although more prevalent, abnormal hemoglobin, creatinine, and glucose values were also not predictive of postoperative adverse outcomes. Routine preoperative testing for hemoglobin, creatinine, glucose, and electrolytes on the basis of age alone may not be indicated in geriatric patients. Rather, selective laboratory testing, as indicated by history and physical examination, which will determine patient's comorbidities and surgical risk, seems to be indicated. IMPLICATIONS: The prevalence of abnormal preoperative electrolyte values and thrombocytopenia was small and had low predictive values. Although more prevalent, abnormal hemoglobin, creatinine, and glucose values were also not predictive of postoperative adverse outcomes. Routine preoperative testing for hemoglobin, creatinine, glucose, and electrolytes on the basis of age alone may not be indicated in geriatric patients. Rather, selective laboratory testing, as indicated by history and physical examination, which will determine patient's comorbidities and surgical risk, seems to be indicated. PMID- 11473850 TI - Designing meaningful industry metrics for clinical productivity for anesthesiology departments. AB - IMPLICATIONS: Clinical productivity measurements that account for differences in clinical settings and concurrencies provided more precise comparisons between two anesthesiology groups. The data show that different concurrencies confound the current industry standard, "per full-time equivalent" measurements, whereas "per operating room site" and "per case" measurements allowed for more meaningful comparisons. PMID- 11473851 TI - Productivity versus availability as a measure of faculty clinical responsibility. AB - Faculty clinical time is an extremely valuable commodity. Most departments quantify faculty clinical time on an "availability" basis (e.g., number of days in the operating room or nights on call). We hypothesize that a productivity measure (i.e., determination of actual clinical care delivered rather than availability of such care) would produce different results than the availability system. The "billable hour" was chosen as the measurement device. It was defined as time that anesthesia was actually given, as obtained from the anesthetic record. After collecting data for a year, we found that despite parity using the availability system, the billable hour system detected significant differences between faculty within and between groups. We conclude that "availability" and "productivity" systems produce different conclusions regarding the relative contributions of an individual faculty or subspecialty group. IMPLICATIONS: Accountability of clinical activities by faculty is crucial to the financial status of any department of anesthesia. We hypothesized that methods of availability (e.g., amount of time scheduled for clinical activities) versus productivity measure (actual amount of clinical care delivered) would be quite different between faculty and differing subspecialty groups. Even though the availability system distributed clinical time on an equal basis, there was a wide difference of clinical productivity within and between specialty groups. We conclude that a productivity measure (i.e., billable hours) is a more accurate reflection of faculty productivity than an availability system and is more in line with departmental sources of financial income. PMID- 11473852 TI - A comparison of minidose lidocaine-fentanyl spinal anesthesia and local anesthesia/propofol infusion for outpatient knee arthroscopy. AB - Traditional methods of spinal anesthesia have proven problematic in the outpatient setting. Minidose lidocaine-fentanyl spinal anesthesia (SAB(MLF)) may be the adaptation necessary to reestablish spinal anesthesia in this venue. One hundred patients scheduled for outpatient knee arthroscopy were randomized to receive either local anesthesia plus a titrated IV propofol infusion (LA/PI) or SAB(MLF) using 20 mg lidocaine 0.5% + 20 microg fentanyl. Patients received midazolam 0.02-0.03 mg/kg IV and fentanyl 0.75-1.0 microg/kg IV upon arrival in the operating room before lumbar puncture or propofol infusion. The propofol infusion was begun at 50-75 microg. kg(-)(1). min(-)(1) and titrated to maintain patient comfort. Boluses (200-400 microg/kg) were given as needed. Local anesthesia included 30 mL lidocaine 1% with epinephrine 1:200,000 intraarticularly plus 10 mL at the portal sites. Three patients (6%) in the LA/PI group versus none in the SAB(MLF) group required general anesthesia. Airway support was required in 54% of the LA/PI patients and in none of the SAB(MLF) patients. Total operating room time (43 vs 45 min), time to home readiness (43 vs 45 min), actual discharge times (73.5 min in both groups), and the incidence of discharge >90 min (22% vs 24%) were the same for both LA/PI and SAB(MLF) groups. LA/PI and SAB(MLF) groups differed in terms of postoperative pruritus (8% vs 68%), pain (44% vs 20%), nausea (8% vs 22%), and ability to void before discharge (56% vs 32%). One patient in each group had mild difficulty initiating voiding at home, but neither required medical attention. In both groups, 90% of patients were either "satisfied" or "very satisfied" with their anesthetic. The two techniques provided comparable patient satisfaction and efficiencies both intraoperatively and in postoperative recovery and discharge. The efficiencies of these techniques were not dependent on special provisions of the physical plant or the practice model. IMPLICATIONS: Both local anesthesia supplemented by a titrated IV propofol infusion and minidose lidocaine-fentanyl spinal anesthesia for outpatient knee arthroscopy provide high patient satisfaction with equally rapid recovery and discharge. PMID- 11473853 TI - The differences in the bispectral index between infants and children during emergence from anesthesia after circumcision surgery. AB - The bispectral index (BIS) correlates with consciousness during adult anesthesia. In this prospective, blinded study of children (n = 24) and infants (n = 25) undergoing elective circumcision, we evaluated BIS and consciousness level during emergence from anesthesia. Anesthesia was maintained with sevoflurane, and a penile nerve block was performed in each patient before surgical stimulation. At the completion of surgery, the sevoflurane was decreased stepwise from 0.9% in increments of 0.2%, and arousal was tested with a uniform auditory stimulus given after a steady state of end-tidal sevoflurane concentration was achieved at each step. The BIS increased significantly as the sevoflurane concentrations decreased in children (0.9%, 62.5 +/- 8.1; 0.7%, 70.8 +/- 7.4; and 0.5%, 74.1 +/- 7.1; P < 0.001 for 0.7% and 0.5% compared with 0.9%), but a similar relationship was not demonstrated in infants. The BIS values at 0.7% and 0.5% sevoflurane were significantly higher in children than infants (P < 0.02 and P < 0.002, respectively). In both children and infants, the BIS increased significantly from pre- to postarousal (children, 73.5 +/- 7 to 83.1 +/- 12, P = 0.01; infants, 67.8 +/- 10 to 85.6 +/- 13.6, P < 0.001). The BIS at which arousal was possible with the stimulus tended to be higher in children than in infants (P = 0.06). IMPLICATIONS: In this study comparing the Bispectral index (BIS) in infants and children undergoing circumcision surgery by use of a standardized surgical and anesthetic technique, a significant decrease in BIS was detected in children during a stepwise decrease in end-tidal sevoflurane concentration. A similar relationship was not demonstrated in infants less than 1 yr old. In both children and infants, BIS increased significantly from pre- to postarousal. Additional studies are necessary to determine changes in BIS with maturational changes in the electroencephalogram. PMID- 11473854 TI - Right internal jugular vein venography in infants and children. AB - We obtained venograms of the right internal jugular vein (RIJV) in 105 infants and children with congenital heart disease during cardiac catheterization. No major anomalies were found in the course of the RIJV. The diameter of the RIJV tended to increase with the patient's age, weight, and height. However, some disproportionately small vessels were seen in 8% of the patients. The depth from the skin to the RIJV varied from 2.5 to 20 mm and did not significantly correlate with age, weight, or height. Confirmation of the diameter or the depth of the RIJV by venography may facilitate clinical decisions and may be useful for performing percutaneous cannulation. IMPLICATIONS: We obtained venograms of the right internal jugular vein in children with congenital heart disease. Generally, the diameter increased with the patient's body size, but disproportionately small vessels were seen in 8% of the patients. Preoperative internal jugular venography may facilitate identifying those patients. PMID- 11473855 TI - Clonidine prevents sevoflurane-induced agitation in children. AB - In a double-blinded trial, 40 male children (age 2-7 yr) undergoing circumcision were randomly assigned to receive clonidine 2 microg/kg IV or placebo after anesthetic induction. For induction and maintenance of anesthesia, we used sevoflurane as the sole anesthetic. For pain treatment, a penile block was performed before surgery. After surgery the incidence and severity of agitation was measured during an observation period of 2 h. Severe agitation was treated with midazolam. In 16 placebo and 2 clonidine-treated patients agitation was observed (P < 0.001). In 6 patients of the Placebo group, agitation was graded as severe, whereas none of the patients in the Clonidine group developed severe agitation (P = 0.02). During the postoperative period heart rate and blood pressure were significantly decreased in clonidine treated patients (P < 0.05). We conclude that clonidine effectively prevents agitation after sevoflurane anesthesia. IMPLICATIONS: The recovery from sevoflurane anesthesia may be complicated by the presence of agitation in pediatric patients. Clonidine 2 microg/kg IV after anesthetic induction effectively reduces the incidence of agitation without resulting in clinically relevant bradycardia and hypotension. PMID- 11473856 TI - The pharmacoeconomics of neuromuscular blocking drugs: a perioperative cost minimization strategy in children. AB - The purpose of this investigation was to compare the costs of intermediate-acting neuromuscular blocking drugs in children during routine ambulatory surgery. We studied 200 healthy, 2-10-yr-old children undergoing elective dental restorative surgery. During Part 1 of the study, children received an inhaled anesthetic with halothane and nitrous oxide, whereas in Part 2, the anesthetic was IV propofol with nitrous oxide. The study drugs were atracurium, cisatracurium, mivacurium, rocuronium, and vecuronium. Patients were initially administered 2x the effective dose for 95% of the study drug. After recovery to 10% of baseline neuromuscular function, the neuromuscular blockade was rigidly maintained with an infusion of the study drug at about 10% of baseline function. Neuromuscular drug costs were approximated as drug usage x cost/unit. The initial drug costs were not substantially different for both Parts 1 and 2, but over time, mivacurium became the most expensive drug and cisatracurium the least expensive. In conclusion, based on current costs, cisatracurium is the least expensive intermediate-acting neuromuscular drug. IMPLICATIONS: For children undergoing minor ambulatory procedures of 1-2 h, and continuous intraoperative neuromuscular blockade is indicated, cisatracurium currently is the least expensive drug. PMID- 11473857 TI - Unmasked residual neuromuscular block after administration of vecuronium for days. AB - IMPLICATIONS: Significant neuromuscular block may be present in patients who have received vecuronium for days. PMID- 11473858 TI - Continuous caudal epidural analgesia for congenital lobar emphysema: a report of three cases. AB - IMPLICATIONS: In congenital lobar emphysema, positive pressure ventilation can expand the emphysematous lobe, compressing the normal lung during anesthesia induction. We managed the dual challenges of safe induction and analgesia for thoracotomy by placing thoracic epidural catheters via the caudal insertion site and retaining spontaneous ventilation until thoracotomy. PMID- 11473859 TI - The influence of inhaled nitric oxide on cerebral blood flow and metabolism in a child with traumatic brain injury. AB - IMPLICATIONS: The effects of inhaled nitric oxide (INO) on cerebrovascular hemodynamics are not well established. We report no adverse cerebral effects with INO therapy in a child with traumatic brain injury. PMID- 11473860 TI - The effects of benzodiazepines on human opioid receptor binding and function. AB - We performed in vitro studies to investigate the potential interaction of benzodiazepines with cloned human opioid receptor subtypes. Midazolam, chlordiazepoxide, and diazepam directly displaced [(3)H]-diprenorphine binding from kappa and delta receptors, but not mu receptors, whereas flumazenil was inactive. These benzodiazepines also stimulated (35)S-GTPgammaS binding in membranes containing human kappa receptors, and the effect of midazolam was prevented by a selective kappa antagonist. Midazolam was also weakly active at delta-receptor activation, whereas all three were inactive at mu receptors. The results suggest that the analgesic efficacy reported for intrathecal benzodiazepines may be attributed, in part, to direct interaction with kappa opioid receptors. IMPLICATIONS: Several human and animal studies have shown analgesic effects of benzodiazepines after spinal injection. Our results show that large concentrations of midazolam, chlordiazepoxide, and diazepam displace the binding of [(3)H]-diprenorphine-an opiate radioligand from kappa receptors. In an in vitro functional assay, midazolam is a weak agonist at the delta-opioid receptor, whereas all three benzodiazepines are kappa-opioid agonists. These findings may partially explain the mechanism of benzodiazepine-induced spinal analgesia. PMID- 11473861 TI - The use of remifentanil to facilitate the insertion of the laryngeal mask airway. AB - Propofol is often used as an IV induction drug for anesthesia and the insertion of a laryngeal mask airway (LMA). As a sole anesthetic, it may be associated with undesirable airway responses such as coughing and gagging. We conducted a randomized, double-blinded study to compare the conditions during insertion of the LMA in 120 patients who received normal saline (Group P), remifentanil 0.25 microg/kg (Group R1), or remifentanil 0.5 microg/kg (Group R2) before the induction of anesthesia with IV propofol. The addition of remifentanil significantly improved the conditions of insertion; in Group R1, 82.5% (33 of 40 patients), and in Group R2, 85.0% (34 of 40 patients) had excellent insertion conditions as compared with the Control group P, 32.5% (13 of 40 patients). Patients in Group P were apneic for a mean (SD) time of 85 (38) s, 186 (75) s in group R1, and 284 (130) s in group R2. There was a lesser decrease in mean arterial blood pressure in group R1. We conclude that remifentanil 0.25 microg/kg, when administered after IV propofol 2.5 mg/kg, provides excellent conditions for insertion of the LMA with minimal hemodynamic disturbances. IMPLICATIONS: Small-dose remifentanil can provide excellent conditions for laryngeal mask airway insertion with minimal hemodynamic disturbances. PMID- 11473862 TI - Clonidine suppresses plasma and cerebrospinal fluid concentrations of TNF-alpha during the perioperative period. AB - The analgesic properties of alpha(2)-agonists are well known. In experimental models, tumor necrosis factor (TNF)-alpha regulates adrenergic responses in the brain. Constitutive TNF-alpha, in brain regions involved in pain perception, is decreased after the administration of clonidine. We investigated patients undergoing lower-extremity revascularization. Seven patients were treated with clonidine 0.2 mg per os (low), and three patients received 0.4 mg per os clonidine (high) before surgery. Eight patients received placebo and served as controls. Continuous spinal anesthesia was provided by insertion of a pliable catheter into the subarachnoid space. Baseline plasma and cerebrospinal fluid (CSF) samples were obtained before injection of local anesthetic. Samples were analyzed for TNF-alpha using a biologic assay. Systemic and central release of catecholamines were assessed by high-pressure liquid chromatography measurement of norepinephrine in plasma and CSF, vanillylmandelic acid and methoxy hydroxyl phenyl glycol in 24-h urinary excretion, respectively. Clonidine 0.2 mg pretreatment decreased TNF-alpha concentrations both in plasma and CSF. Patients receiving clonidine had lower pain visual analog scale scores and required less morphine compared with the Placebo group (P < 0.01). Preoperative administration of clonidine decreased catecholamine release in the periphery, as well as in the central nervous system. A smaller norepinephrine concentration in plasma and CSF, and less secretion of vanillylmandelic acid (P < 0.01) and methoxy hydroxyl phenyl glycol in the urine, were observed. Larger dose clonidine (0.4 mg) resulted in no detectable TNF-alpha in CSF. These results suggest that an interaction between TNF-alpha and the function of adrenergic neurons in the central nervous system may contribute to the sedative and analgesic effects of adrenergic agonists. IMPLICATIONS: Preoperative administration of clonidine decreases both plasma and cerebrospinal fluid concentrations of inflammatory cytokines, resulting in perioperative analgesia and decreased sympathetic tone. PMID- 11473863 TI - A multicenter evaluation of the time-course of action of two doses of rapacuronium after early and late reversal with neostigmine. AB - Early reversal of rapacuronium may accelerate return of neuromuscular function. This study was designed to compare early (2 min after rapacuronium) or late (at 25% recovery of the first twitch [T1] of train-of-four) reversal of rapacuronium with neostigmine. We studied 119 subjects between the ages of 18 and 75 yr. Anesthesia was induced with fentanyl and thiopental and maintained with nitrous oxide, propofol, and fentanyl. Mechanomyographic neuromuscular monitoring was performed by using train-of-four stimulation of the ulnar nerve. Two groups received 1.5 mg/kg rapacuronium followed by neostigmine (50 microg/kg) and glycopyrrolate (10 microg/kg) either at 2 min after rapacuronium bolus or at 25% T1 recovery. The other two groups received 2.0 mg/kg rapacuronium, after which neostigmine was similarly given. For each rapacuronium dose, the time from the administration of rapacuronium to the start of T1 recovery or 25% T1 recovery was significantly shorter in subjects who received the reversal 2 min after rapacuronium. However, late recovery, defined by times from administration of rapacuronium to 70%, or 80% T4/T1 recovery, was not influenced by early reversal administration. We conclude that initial recovery is accelerated by early administration of neostigmine. Time to full recovery after rapacuronium administration is, however, dose-dependent and not significantly altered by early administration of neostigmine. IMPLICATIONS: "Rescue reversal," which includes the administration of neostigmine shortly after the administration of rapacuronium, may accelerate the return of spontaneous breathing (early recovery), but does not shorten the time to complete recovery of upper airway function. PMID- 11473864 TI - The effects of isoflurane on native and chimeric muscarinic acetylcholine receptors: the role of protein kinase C. AB - By using two electrode voltage clamps, we investigated the effects of isoflurane on m3 and chimeric m1/m3 muscarinic receptors and the role of protein kinase C (PKC) in the effects. Muscarinic receptors were expressed by injection of mRNA into Xenopus oocytes, and Ca(2+)-activated Cl(-) currents were measured after the application of acetyl-beta-methylcholine. We constructed chimeric m1/m3 receptor DNA encoding the third intracellular loop of m1 and the remainder from the m3 receptor. Chimeric and m3 receptors were inhibited by isoflurane, but the m1 receptor was not. PKC activation with phorbol-12-myrisate-13-acetate (50 nM) decreased signaling of both chimeric and m3 receptors significantly. Chelerythrine (20 microM, PKC inhibitor) abolished the effect of isoflurane on chimeric and m3 signaling. Whereas isoflurane inhibition of chimeric and m3 receptors was completely reversible after washout with Tyrode's solution for 3 min, treatment with okadaic acid (500 nM, protein phosphatase inhibitor) rendered the inhibition irreversible. Taken together, our results suggest that isoflurane inhibits m3 and chimeric m1/m3 muscarinic signaling by enhancing PKC activity and that the site of action is located outside of the third intracellular loop. IMPLICATIONS: By use of the Xenopus oocyte expression system, we investigated the effects of isoflurane on muscarinic signaling and the role of protein kinase C in these effects. Our findings suggest that isoflurane inhibits muscarinic receptors through activation of protein kinase C and that the relevant phosphorylation sites are located outside the third intracellular loop. PMID- 11473865 TI - Propofol in an emulsion of long- and medium-chain triglycerides: the effect on pain. AB - IMPLICATIONS: In a test of two formulations of propofol for induction, patients experienced less pain with the formulation in Intralipid (Propofol-Lipuro 1%) than with Diprivan 1%. PMID- 11473866 TI - A new, safety-oriented, integrated drug administration and automated anesthesia record system. AB - Medication errors are an important cause of patient morbidity and mortality and excessive costs, including in anesthesia. Conventional methods of injectable drug administration in anesthesia make little use of technology to support manual checking and are idiosyncratic and relatively error prone. Similarly, conventional anesthesia records are handwritten, time-consuming to make, and often unreliable. There are automated record systems, but they do not provide support for checking drugs. Therefore, by using a multifaceted approach based on established principles of systems design and human factors psychology, we have developed a system that includes trays that promote a well-organized anesthetic workspace, color- and bar-coded labeling of syringes, and automatic visual and auditory verification of the syringe labels by computer just before each drug administration. In addition, documentation of drugs administered and a traditional anesthetic case record are generated automatically. The system has been successfully deployed for 25 mo and has been used by 35 anesthesiologists in 1148 diverse cases, including cardiopulmonary bypass procedures, heart and lung transplants, and orthopedic and otorhinolaryngologic operations. It is in daily use in a tertiary teaching center and in a private hospital. IMPLICATIONS: Traditional methods of drug administration and record keeping in anesthesia are relatively error prone. By using sound principles of systems design and human factors psychology, we have designed and deployed a system with the aim of improving patient safety by facilitating correct drug administration and accurate anesthesia record making. PMID- 11473867 TI - The ADU vaporizing unit: a new vaporizer. AB - We determined the performance of the vaporizer of the ADU machine (Anesthesia Delivery Unit; Datex-Ohmeda, Helsinki, Finland). The effects of carrier gas composition (oxygen, oxygen/N(2)O mixture, and air) and fresh gas flow (0.2 to 10 L/min) on vaporizer performance were examined with variable concentrations of isoflurane, sevoflurane, and desflurane across the whole range of each vaporizer's output. In addition, the effects of sudden changes in fresh gas flow and carrier gas composition, back pressure, flushing, and tipping were assessed. Vaporizer output depended on fresh gas flow, carrier gas composition, dial settings, and the drug used. Vaporizer output remained within 10% of dial setting with fresh gas flows of 0.3-10 L/min for isoflurane, within 10% of dial setting with fresh gas flows of 0.5-5 L/min for sevoflurane, and within 13% of dial setting with fresh gas flows of 0.5 to 1 L/min for desflurane. Outside these fresh gas flow ranges, output deviated more. The effect of sudden changes in fresh gas flow or carrier gas composition, back pressure, flushing, and tipping was minimal. We conclude that the ADU vaporizer performs well under most clinical conditions. Despite a different design and the use of complex algorithms to improve accuracy, the same physical factors affecting the performance of conventional vaporizers also affect the ADU vaporizer. IMPLICATIONS: The ADU vaporizer performs well under most clinical conditions. Despite a different design and the use of complex algorithms to improve accuracy, the same physical factors affecting the performance of conventional vaporizers also affect the ADU vaporizer. PMID- 11473868 TI - Intraoperative monitoring of the recurrent laryngeal nerve in 151 consecutive patients undergoing thyroid surgery. AB - IMPLICATIONS: We present a technique of intraoperative monitoring of the recurrent laryngeal nerve using a surface electrode attached to a routine endotracheal tube. The technique proved noninvasive, easy to use, and reliable in 151 prospective consecutive patients for preventing permanent laryngeal nerve damage in thyroid surgery. PMID- 11473869 TI - The pharmacokinetics of cisatracurium in patients with acute respiratory distress syndrome. AB - Continuous neuromuscular blockade is often necessary in patients being treated for acute respiratory distress syndrome (ARDS) to optimize oxygenation. In this study, neuromuscular blockade (no response to two responses at the train-of-four stimulation at the orbicularis oculi muscle) was achieved in six patients with ARDS by a continuous infusion of cisatracurium. The plasma concentration of cisatracurium during the infusion averaged 1.00 (0.25-1.45) microg/mL, expressed as median (range). The clearance and half-life were 6.5 (3.3-7.6) mL. min(-1). kg(-1) and 25 (16-48) min, respectively. The laudanosine plasma concentrations were 0.70 (0.12-1.20) microg/mL. The pharmacokinetic variables of cisatracurium are similar to those of patients without organ failure undergoing elective surgery. Plasma laudanosine levels always remained well less that those associated with seizure activity in animal models. Long-term infusion of cisatracurium was not associated with any side effects. Cisatracurium is a suitable muscle relaxant when deep and continuous levels of muscle relaxation are required in patients treated for ARDS. IMPLICATIONS: We studied the pharmacokinetics of cisatracurium in six patients treated for respiratory distress syndrome by continuous muscle relaxation. A deep degree of neuromuscular blockade corresponding to abolition of two responses at the orbicularis oculi to train-of-four stimulation was obtained in all patients. The pharmacokinetic variables observed in these severely ill patients were similar to those of anesthetized patients. No accumulation of laudanosine was seen. Cisatracurium appears to be suitable when continuous muscle relaxation is required in critically ill patients. PMID- 11473870 TI - Colloids versus crystalloids and tissue oxygen tension in patients undergoing major abdominal surgery. AB - The effects of intravascular volume replacement regimens on tissue oxygen tension (ptiO(2)) are not definitely known. Forty-two consecutive patients scheduled for elective major abdominal surgery were prospectively randomized to receive either 6% hydroxyethyl starch (HES) (mean molecular weight 130 kd, degree of substitution 0.4, n = 21) or lactated Ringer's solution (RL, n = 21) for intravascular volume replacement. Fluids were administered perioperatively and continued for 24 h on the intensive care unit to keep central venous pressure between 8 and 12 mm Hg. The ptiO(2) was measured continuously in the left deltoid muscle by using microsensoric implantable partial pressure of oxygen catheters after the induction of anesthesia (baseline, T0), 60 min (T1) and 120 min thereafter (T2), at the end of surgery (T3), and on the morning of the first postoperative day on the intensive care unit (T4). HES 130/0.4 2920 +/- 360 mL and 11,740 +/- 2,630 mL of RL were given to the patients within the study period. Systemic hemodynamics and oxygenation (PaO(2), PaCO(2)) did not differ significantly between the two volume groups throughout the study. From similar baseline values, ptiO(2) increased significantly in the HES-treated patients (a maximum of 59% at T4), whereas it decreased in the RL group (a maximum of -23% at T4, P < 0.05). The largest differences of ptiO(2) were measured on the morning of the first postoperative day. We conclude that intravascular volume replacement with 6% HES 130/0.4 improved tissue oxygenation during and after major surgical procedures compared with a crystalloid-based volume replacement strategy. Improved microperfusion and less endothelial swelling may be responsible for the increase in ptiO(2) in the HES 130/0.4-treated patients. IMPLICATIONS: In patients undergoing major abdominal surgery, a colloid-based (with hydroxyethyl starch [HES] 130/0.4) and a crystalloid-based (with lactated Ringer's solution [RL]) volume replacement regimen was compared regarding tissue oxygen tension (ptiO(2)) measured continuously by microsensoric implantable catheters. The ptiO(2) increased in the HES-treated (+59%) but decreased in the RL-treated ( 23%) patients. Improved microcirculation may be the mechanism for the better ptiO(2) in the HES group. PMID- 11473871 TI - The effect of intrathecal analgesia on the success of external cephalic version. AB - External cephalic version (ECV), the procedure whereby a fetus in the breech position is converted to vertex, is often performed to avoid an operative delivery. Potential benefits of epidural and spinal anesthesia for this procedure are controversial. Several previous studies have evaluated the use of epidural anesthesia with varying results. We sought to determine whether analgesia produced by subarachnoid sufentanil would safely improve the success of ECV. Patients who received subarachnoid analgesia (n = 20) were compared with those who did not (n = 15) in regard to success of ECV, level of pain during ECV, and satisfaction. ECV was successful in 21 patients (60%), with more frequent success in women who received spinal analgesia as compared with those who did not (80% vs 33%, respectively; P = 0.005). Patients who received spinals also reported smaller pain scores and were more satisfied with ECV. None of the women who received spinal analgesia developed a postdural puncture headache, and the only case of fetal bradycardia occurred in a patient who did not receive spinal analgesia. More profound patient comfort after spinal analgesia may have permitted greater manipulation of the abdomen during ECV, thus improving success rates of ECV without increasing risk. IMPLICATIONS: The success of external cephalic version (ECV) was compared in women who received spinal analgesia and those who did not. Successful ECV occurred more frequently in those women who received spinal analgesia. Because term singleton pregnancies associated with breech position usually require cesarean delivery, an increase in success of ECV may decrease the number of cesarean deliveries performed. PMID- 11473872 TI - Predictors of breakthrough pain during labor epidural analgesia. AB - Parturients who receive labor epidural analgesia may experience breakthrough pain that requires supplemental medications. We investigated the factors associated with breakthrough pain. This prospective observational study included 1963 parturients who received epidural analgesia. Subjects were categorized into two groups on the basis of the number of episodes of breakthrough pain: the Recurrent Breakthrough Pain (RBP) group experienced three or more episodes. Univariate and multivariate regression analyses were used to evaluate factors associated with the RBP group. By multivariate analysis, nulliparity, heavier fetal weight, and epidural catheter placement at an earlier cervical dilation were found to be independently associated with the RBP group. These factors may predict which parturients' analgesia may be complicated by breakthrough pain. Parturients who received a combined spinal/epidural technique were less likely to be associated with the RBP group. The combined spinal/epidural technique may be superior to conventional epidural anesthesia, because breakthrough pain occurred less often. It is interesting to note that the characteristics that are associated with the RBP group are similar to those that have been associated with increased severity of maternal pain. IMPLICATIONS: Nulliparity, heavier fetal weight, and epidural catheter placement at an early cervical dilation are predictors of breakthrough pain during epidural labor analgesia. The combined spinal/epidural technique may be associated with a decreased incidence of breakthrough pain. PMID- 11473873 TI - The effects of capsaicin cream on prostaglandin-induced allodynia. AB - Although intradermal injection of capsaicin produces acute pain and secondary hyperalgesia, long-term topical application of capsaicin cream has been used as a medication for pain relief in various pain conditions. We previously reported that intrathecal administration of prostaglandin (PG) E(2) and PGF(2alpha) into mice induced touch-evoked pain (allodynia) through capsaicin-sensitive and capsaicin-insensitive afferent fibers, respectively. To clarify the mechanism of an analgesic effect by capsaicin cream, here we applied it to the tail and hind paws of mice and investigated its effects on PGE(2)- and PGF(2alpha)-induced allodynia. Twenty-four-hour pretreatment of mice with 0.025% or 0.05% capsaicin cream markedly alleviated allodynia induced by PGE(2), but not by PGF(2alpha). These results suggest that the topical application of capsaicin cream modulates capsaicin-sensitive afferents and ameliorates allodynia evoked by PGE(2) at the spinal level. IMPLICATIONS: Topical application of capsaicin cream alleviates touch-evoked pain induced by the intrathecal administration of prostaglandin E(2). This study may provide a rationale for the use of capsaicin cream as a therapeutic drug for pain relief. PMID- 11473874 TI - The residual effects of hemorrhagic shock on pain reaction and c-fos expression in rats. AB - To investigate the residual effects of hemorrhagic shock on pain reaction and c fos expression, we performed formalin tests after hemorrhage and reinfusion in rats. Twenty adult male Sprague-Dawley rats were divided into Control (n = 10) and Postshock (n = 10) groups. The mean blood pressure of the Control group was 100-120 mm Hg, and that of the Postshock group was kept at 50-60 mm Hg for 30 min by draining blood. After 15 min of returning mean blood pressure to normal levels in the Postshock group, 10% formalin (3.7% formaldehyde solution, 100 microL) was injected into the left rear paw of both groups. Nociceptive behaviors were observed for 1 h after the formalin injection. The rats were killed at 2 h after the formalin injection, and the lumbar spinal cord was then stained for c-fos immunohistochemistry by using the avidin-biotin-peroxidase method. Animals in the Postshock group showed considerably less nociceptive behavior than those in the Control group. C-fos expression in the deep layer (IV-VI) of the spinal cord was significantly less in the Postshock group. In conclusion, decreases of nociceptive behaviors and c-fos expression were observed under normotensive conditions after hemorrhagic shock. The mechanisms governing these reactions remain unclear. IMPLICATIONS: Formalin tests were performed after hemorrhage and reinfusion in rats. A stress-induced analgesia was observed under normotensive conditions after hemorrhagic shock. The mechanisms remain unclear. PMID- 11473875 TI - An isobolographic analysis of the adrenergic modulation of diclofenac antinociception. AB - We evaluated the noradrenergic modulation of the antinociceptive activity of diclofenac in mice using the acetic acid writhing test. Dose-response curves were obtained for the antinociceptive effect of diclofenac, phenylephrine, clonidine, desipramine, prazosin, and yohimbine administered both systemically and intrathecally, and ED(50)s were calculated. Noradrenergic modulation was evaluated by performing an isobolographic analysis of the systemic or intrathecal coadministration of fixed-ratio combinations of diclofenac with each adrenergic drug. The systemic, but not the intrathecal, combinations of diclofenac with phenylephrine or clonidine showed supraadditivity, suggesting that the activation of alpha(1) and alpha(2) adrenoceptors interfered with the nociceptive transmission at spinal and supraspinal levels. Supraadditive effects were not demonstrated for the intrathecal injection of diclofenac combined with phenylephrine, clonidine and a selective norepinephrine uptake inhibitor (desipramine) or adrenergic antagonists. We conclude that interaction between adrenoceptors and diclofenac can modulate antinociception by activating common or different mechanisms. Diclofenac has an antinociceptive activity that, in addition to cyclooxygenase inhibition, can be modulated by additive and supraadditive interactions with adrenergic drugs. IMPLICATIONS: Diclofenac analgesia in mice can be modulated by interaction with adrenergic drugs. The systemic but not the intrathecal administration of phenylephrine and clonidine produced supraadditive interactions. For desipramine, prazosin, and yohimbine, supraadditive interactions were not statistically demonstrated. The coadministration of drugs inducing supraadditive effects could be clinically relevant for the treatment of chronic pain because of reduction of doses and side effects. PMID- 11473876 TI - An evaluation of the infraclavicular block via a modified approach of the Raj technique. AB - Infraclavicular plexus block has recently become a technique of increasing interest. However, no approach has provided easily identifiable landmarks, good conditions for catheter placement, and lack of complications (mainly pneumothorax). We describe a modified approach of the Raj technique based on the identification of the anterior acromial process, jugular notch, and emergence of the axillary artery within the axillary fossa, with the arm abducted to 90 degrees and elevated by approximately 30 degrees. We evaluated the clinical characteristics of this approach by injecting 40 to 50 mL of ropivacaine 0.6% in 150 patients scheduled for elective surgery of the forearm, wrist, or hand. Success was defined as a sensory block of the 5 nerves with territories distal to the elbow within 30 min after performing the block. The success rate was 97% when a distal response (flexion or extension of the wrist or fingers) was elicited and 44% when a proximal (contraction of the triceps, biceps) was obtained using a nerve stimulator. Complications were rare: aspiration of blood was seen in 2% of patients and hematoma was seen at the puncture site in 0.6%; no pneumothorax occurred. Eleven patients (7%) complained of some pain during the procedure. We conclude that the modified approach of the Raj technique for infraclavicular block is very effective when a distal nerve stimulator response is obtained with a small complication rate and a high degree of patient satisfaction. IMPLICATIONS: We describe a modified approach of the Raj technique for the infraclavicular brachial plexus. The elicitation of a distal nerve stimulator response is associated with a high success rate, a low incidence of complications and a high degree of patient satisfaction. PMID- 11473877 TI - The supraclavicular lateral paravascular approach for brachial plexus regional anesthesia: a simulation study using magnetic resonance imaging. AB - In the supraclavicular lateral paravascular approach for brachial plexus regional anesthesia by Moorthy et al. (Moorthy's block), the patient is supine with the ipsilateral shoulder displaced anteriorly 5-8 cm. The needle direction is precisely defined in the coronal plane (using a Doppler flowprobe) but not in the sagittal plane. We sought to determine whether the block could be simplified by keeping the shoulder in a neutral position, if the needle direction in the sagittal plane could be more precisely described, and if the risk of pneumothorax appeared acceptably small. These questions were studied by magnetic resonance imaging in 10 volunteers. Volume datasets of the periclavicular region allowed precise positioning of simulated needles. In all volunteers, Moorthy's block could be performed with the shoulder in a neutral position. The optimal needle trajectory passed 5 mm posterior to the clavicle and was 25 degrees posterior to the coronal plane, never approaching the pleura closer than 18 mm. We conclude that Moorthy's block can be performed with the shoulder in a neutral position, that more precise instructions for the needle direction can be given, and that the risk of pneumothorax seems minimal. This should be confirmed by a clinical study. IMPLICATIONS: We studied an established method for brachial plexus block with needle advancement in the chest region in volunteers using magnetic resonance imaging. Our results suggest a simplification of the method and more guidelines for the needle angle to the skin, with a minimal risk for lung injury. PMID- 11473878 TI - The pharmacokinetics and pharmacodynamics of bupivacaine-loaded microspheres on a brachial plexus block model in sheep. AB - We evaluated bupivacaine-loaded microspheres (B-Ms) using a brachial plexus block model in sheep. In the first step, pharmacokinetic characterization of 75 mg bupivacaine hydrochloride (B-HCl) (IV infusion and brachial plexus block) was performed (n = 12). In the second step, a brachial plexus block dose response study of B-HCl was performed with 37.5 mg, 75 mg, 150 mg, 300 mg, and 750 mg. As a comparison, evaluations were performed using a 750-mg bupivacaine base (B). In the third step, evaluations of brachial plexus block were performed with B-Ms (750 mg of B as B-Ms) using two formulations, 60/40 and 50/50 (w/w %); drug-free microspheres were also evaluated. Toxicity evaluations were also performed after IV administration of B-HCl (750 mg and 300 mg), B-Ms (750 mg), and drug-free microspheres (30 mL over 1 min). As the B-HCl dose increased, the time of onset of block decreased and the duration of complete motor blockade increased at the expense of an increase in bupivacaine plasma concentrations. The time of maximum concentration appeared to be independent of the B-HCl dose. In brachial plexus block, a 37.5-mg dose of B-HCl did not induce motor blockade whereas a dose of 750 mg of B-HCl was clinically toxic. In the case of IV administration, doses of 300 mg of B-HCl were as toxic as 750 mg of B-HCl. Compared with the 75 mg of B HCl administration for brachial plexus block, administration of 750 mg of B as B Ms increased the duration of complete motor blockade without significant difference in maximum concentration. No significant clinical difference between the two formulations of B-Ms was demonstrated. The IV administration of B-Ms was safe. We conclude that the controlled release of bupivacaine from microspheres prolonged the brachial plexus block without obvious toxicity. IMPLICATIONS: Administration of 750 mg of bupivacaine as loaded-microspheres resulted in prolongation of brachial plexus block in sheep. The peak plasma concentration was not significantly larger than that obtained with 75 mg of plain bupivacaine. The motor blockade was increased more than six times compared with 75 mg plain bupivacaine. PMID- 11473879 TI - The effect of large-dose intrathecal opioids on the autonomic nervous system. AB - Decreases in blood pressure after the spinal injection of opioids suggest that intrathecal (IT) opioids may have a sympatholytic effect similar to that of local anesthetic drugs. We compared two groups of patients aged 10-16 yr (n = 10 in each group). Group One (IT group) received IT opioids. Group Two (Epidural group) received 0.5% bupivacaine epidurally. The sympathetic effects of IT opioids and epidural bupivacaine were monitored by the changes in toe relative to calf temperature and by the changes in pulse wave gradients with digital plethysmography. Changes in temperature gradients comparing calf to toe and increases in pulse amplitude indicate vasodilatation caused by sympathetic blockade in this model. Calf to toe temperature gradients (Deltacalf-Deltatoe) were evaluated by subtracting the two measurements. Pulse wave plethysmography was recorded before and after spinal and epidural injection at intervals of 10 min for 40 min. All patients demonstrated changes in their calf to toe gradients after IT and epidural injections (-3.2 +/- 1.6). Systolic blood pressure decreased from a mean of 70 +/- 15 mm Hg to 55 +/- 10 mm Hg. Pulse wave plethysmography amplitude increased after the intrathecal opioid and epidural bupivacaine injection similarly. We conclude that the increases in pulse wave amplitude and decreases in calf-toe gradients indicate a sympatholytic effect after IT opioids similar to that of local anesthetics. IMPLICATIONS: The sympatholytic effects of neuraxial opioids were compared with those of local anesthetics. Two groups of patients were assigned to receive a neuraxial opioid or bupivacaine. Our results demonstrate that opioids cause hypotension and peripheral vasodilatation similar to bupivacaine. This finding suggests that neuraxial opioids have a sympatholytic effect comparable to that of local anesthetic drugs. PMID- 11473880 TI - The in vivo effects of general and epidural anesthesia on human immune function. AB - Impaired in vivo immunity is often observed after major surgery and is multifactorial. We conducted a randomized clinical study to determine the independent effects of general anesthesia (GA) and of lumbar epidural anesthesia (LEA) on human immune function in the absence of surgical trauma. Nineteen healthy volunteers were randomized to receive GA with thiopental and isoflurane, LEA with lidocaine, or no anesthesia (Control). Serial blood samples were tested for antibody responses to antigen inoculation, neutrophil and mononuclear cell antibody-dependent cell cytotoxicity (ADCC), natural killer cell cytotoxicity, and neutrophil phagocytic activity. Antibody responses were similar in the three groups. Mononuclear cell ADCC increased in the LEA group at the end of the anesthetic (P < 0.05 at effector/target [E/T] ratios of 10:1, 25:1, and 50:1). Natural killer cell cytotoxicity increased at the end of the anesthetic in both the LEA group (P < 0.05 at all E/T ratios) and the GA group (P < 0.05 at an E/T ratio of 5:1 and 10:1). No significant changes were observed for neutrophil ADCC or phagocytosis. General or epidural anesthesia alone, in the absence of surgery, seems to have only transient and minor effects on human immune function. IMPLICATIONS: General or epidural anesthesia alone, in the absence of surgery, seems to have only transient and minor effects on human immune function. PMID- 11473881 TI - Topical application of acidic bupivacaine to the lumbar ganglion induces mechanical hyperalgesia in the rat. AB - To investigate the neurologic mechanisms of acidic local anesthetic-induced low back pain in humans, we administered bupivacaine and buffered saline at acidic or alkalinized pH at the L5 dorsal root ganglion (DRG) of rats via a hole drilled through the transverse process covering the DRG. Behavioral changes were tested before and after bupivacaine or saline administration. Results indicate that acute single-dose infusion of the DRG with bupivacaine (0.5%) at acidic pH (5.5) induced ipsilateral mechanical hyperalgesia that lasted for 7 days. Acute infusion of alkalinized bupivacaine (pH 7.2), however, caused only minor hyperalgesia that lasted <3 days. Similar results were obtained when bupivacaine was replaced with saline. Alternatively, chronic delivery of acidic saline to the DRG via a subcutaneously implanted osmotic pump resulted in a significant decrease in the withdrawal threshold on the ipsilateral hind paw that lasted for 10 days. In rats receiving chronic treatment of the DRG with alkalinized saline, mechanical hyperalgesia lasted for only 3 days. The results demonstrated that acidic bupivacaine deposited at the DRG causes pain and hyperalgesia when the effects of the local anesthetic have dissipated. These findings may explain the limited therapeutic effects of some acidic local anesthetics used for management of cancer-related and chronic back pain. IMPLICATIONS: Acidic bupivacaine administered at the L5 lumbar ganglion causes pain and hypersensitivity of the hind paw in the rat. These findings may explain the limited therapeutic effects of some acidic local anesthetics used for treatment of cancer-related and chronic back pain. PMID- 11473882 TI - More epidural than intravenous sufentanil is required to provide comparable postoperative pain relief. AB - The extent to which epidurally administered sufentanil acts directly on spinal opioid receptors remains controversial. We tested the hypothesis that small-dose boluses of sufentanil, given epidurally or IV, provide comparable analgesia at similar plasma sufentanil concentrations. The lipophilicity of sufentanil makes it likely to be absorbed into fat surrounding the epidural space. We therefore also tested the hypothesis that more epidural than IV sufentanil is required to produce comparable analgesia. Analgesia and plasma sufentanil concentrations were evaluated in 20 postoperative patients randomly assigned to patient-controlled epidural or IV sufentanil. Pain was evaluated with visual analog scales by blinded observers. Sufentanil doses and plasma concentrations were measured. Analgesia was similar with epidural and IV sufentanil administration. Plasma sufentanil concentrations were virtually identical in the two groups. However, significantly larger sufentanil doses were required with epidural administration: 238 +/- 50 microg vs 160 +/- 32 microg (P < 0.01). The primary mechanism by which small-dose boluses of epidurally-administered sufentanil produce analgesia seems to be systemic absorption of the drug with subsequent recirculation to the supraspinal opioid receptors. This study demonstrates that the cumulative dose of sufentanil, when administered as a small epidural bolus, is approximately 50% more than that administered IV to provide comparable analgesia. This indicates that the bioavailability of epidurally-administered sufentanil is reduced and suggests that a large proportion of the drug may be absorbed into the epidural fat. IMPLICATIONS: More epidural than IV sufentanil was required to provide comparable postoperative pain relief and similar plasma sufentanil concentrations. These data suggest that when sufentanil is administered in small dose boluses, much of the drug is absorbed into the epidural fat and that the primary mechanism by which epidurally administered sufentanil produces analgesia is via systemic absorption. PMID- 11473883 TI - A study of the paravertebral anatomy for ultrasound-guided posterior lumbar plexus block. AB - IMPLICATIONS: We investigated the feasibility of posterior paravertebral sonography as a basis for ultrasound-guided posterior lumbar plexus blockades. Posterior paravertebral sonography proved to be a reliable as well as accurate imaging procedure for visualization of the lumbar paravertebral region except the lumbar plexus. PMID- 11473884 TI - The analgesic efficacy of patient-controlled bupivacaine wound instillation after total abdominal hysterectomy with bilateral salpingo-oophorectomy. AB - To assess the effect of local anesthetic wound instillation on visceral and somatic pain, we studied 36 patients undergoing total abdominal hysterectomy and bilateral salpingo-oophorectomy. A standard general anesthetic was administered. On completion of the operation, a multiorifice 20-gauge epidural catheter was placed above the superficial abdominal fascia such that the tip was at the midpoint of the surgical wound. After surgery, either bupivacaine 0.25% (Bupivacaine group) or sterile water (Control group) was administered via a patient-controlled analgesia device programmed to deliver 9.0 mL with a 60-min lockout interval. During the first 6 h after surgery, rescue IV morphine (2 mg) was administered every 10 min until a visual analog scale score of <30 mm was achieved. Thereafter, on patient request, rescue meperidine 1 mg/kg IM was administered. When compared with the Control group, significantly (P < 0.001) less rescue analgesia was administered to patients in the Bupivacaine group. Rescue morphine administered during the first 6 h after surgery was 6 +/- 4 mg versus 12 +/- 6 mg (P < 0.001) for the Bupivacaine and Control groups, respectively. Rescue meperidine administered was 29 +/- 37 mg versus 95 +/- 36 mg (P < 0.001) for the Bupivacaine and Control groups, respectively. Nausea and antiemetic drug administration was significantly (P = 0.003) less in the Bupivacaine group. Pain scores were similar between the groups. Patient satisfaction was significantly (P = 0.04) more in the Bupivacaine group. We conclude that bupivacaine wound instillation decreases opioid requirements and nausea in the first 24 h after total abdominal hysterectomy with bilateral salpingo-oophorectomy. IMPLICATIONS: Bupivacaine instillation via an electronic patient-controlled analgesia device provides effective analgesia after total abdominal hysterectomy with bilateral salpingo-oophorectomy. PMID- 11473885 TI - Carbon monoxide formation in dry soda lime is prolonged at low gas flow. AB - When exposed to volatile anesthetics containing a CHF(2)-group, such as isoflurane, desiccated absorbents produce carbon monoxide (CO). In the anesthesia circuit, the anesthetic flow that passes through the absorber varies with the minute ventilation. We sought to determine CO formation at different levels of test gas flow. Isoflurane 0.5% (series A) or 0.5% isoflurane + 5% CO(2) (series B) in pure O(2) was passed through dry soda lime samples (600 g, Draegersorb 800((R))) at flows of 1, 3, 5, 7, and 10 L/min. Each experiment was performed three times. At the outlet, CO concentration, isoflurane concentration and temperature were continuously recorded. In both series, the duration of CO formation was inversely related to the test gas flow. In series B, mean CO concentrations and the calculated amount of CO formation decreased significantly with increasing flow rates, which was not the case in series A. In both series, the higher the flow rate, the higher was the temperature and the shorter the time until the isoflurane concentration increased to the set level. We conclude that anesthetic degradation in dry soda lime is finite and, as long as no CO(2) is added, will produce roughly the same amount of CO regardless of inflow rate. The inflow rate influences the duration of CO formation such that at lower minute ventilation longer CO formation can be expected. IMPLICATIONS: CO formation from isoflurane degradation in dry soda lime was determined at different rates of test gas flow. The duration and, in the presence of CO(2), the total amount of CO formation were inversely related to the flow rate. PMID- 11473886 TI - Gastroesophageal reflux and aspiration of gastric contents in anesthetic practice. AB - General anesthesia may predispose patients to aspiration of gastroesophageal contents because of depression of protective reflexes during loss of consciousness. In addition, some patients may be at increased risk of pulmonary aspiration because of retention of gastric contents caused by pain, inadequate starvation, or gastrointestinal pathology resulting in reduced gastric emptying and gastroesophageal reflux. Despite increasing knowledge of the problems associated with aspiration, the relatively small incidence and associated mortality rates in the perioperative period do not appear to have changed markedly over the last few decades. In this review article, the physiological factors associated with an increased risk of gastroesophageal reflux and aspiration are considered together with some of the methods that are used to prevent aspiration. In particular, preoperative starvation, the use of drugs designed to increase gastric pH, recent developments in airway devices, and appropriate application of cricoid pressure are critically appraised. PMID- 11473887 TI - Whose value is it anyway? PMID- 11473888 TI - A randomized controlled trial on local anesthetics in the wound: what do the numbers mean? PMID- 11473889 TI - Excessive voltage output? PMID- 11473890 TI - Carbon monoxide rebreathing during low flow anesthesia. PMID- 11473892 TI - Thromboelastography: another point of view. PMID- 11473891 TI - Prophylactic use of tranexamic acid and incidence of arterial thrombosis in liver transplantation. PMID- 11473893 TI - Postoperative nausea and vomiting in female patients. PMID- 11473894 TI - Myocardial ischemia in cataract surgery patients. PMID- 11473895 TI - Policies to tackle social exclusion. PMID- 11473896 TI - Reaching all children. PMID- 11473897 TI - Social capital. PMID- 11473898 TI - Disability discrimination. PMID- 11473899 TI - One Bristol, but there could have been many. PMID- 11473900 TI - Bristol inquiry condemns hospital's "club culture". PMID- 11473901 TI - Individuals who should have behaved differently. PMID- 11473902 TI - Kennedy refused to read the General Medical Council's reports. PMID- 11473903 TI - Kyoto protocol rescued in Bonn. PMID- 11473904 TI - Summit fails to agree new deal on world debt. PMID- 11473905 TI - Oxfam accuses Pfizer of "moral bankruptcy". PMID- 11473906 TI - Health effects of housing improvement: systematic review of intervention studies. AB - OBJECTIVE: To review the evidence on the effects of interventions to improve housing on health. DESIGN: Systematic review of experimental and non-experimental housing intervention studies that measured quantitative health outcomes. DATA SOURCES: Studies dating from 1887, in any language or format, identified from clinical, social science, and grey literature databases, personal collections, expert consultation, and reference lists. MAIN OUTCOME MEASURES: Socioeconomic change and health, illness, and social measures. RESULTS: 18 completed primary intervention studies were identified. 11 studies were prospective, of which six had control groups. Three of the seven retrospective studies used a control group. The interventions included rehousing, refurbishment, and energy efficiency measures. Many studies showed health gains after the intervention, but the small study populations and lack of controlling for confounders limit the generalisability of these findings. CONCLUSIONS: The lack of evidence linking housing and health may be attributable to pragmatic difficulties with housing studies as well as the political climate in the United Kingdom. A holistic approach is needed that recognises the multifactorial and complex nature of poor housing and deprivation. Large scale studies that investigate the wider social context of housing interventions are required. PMID- 11473907 TI - Financial cost of social exclusion: follow up study of antisocial children into adulthood. AB - OBJECTIVES: To compare the cumulative costs of public services used through to adulthood by individuals with three levels of antisocial behaviour in childhood. DESIGN: Costs applied to data of 10 year old children from the inner London longitudinal study selectively followed up to adulthood. SETTING: Inner London borough. PARTICIPANTS: 142 individuals divided into three groups in childhood: no problems, conduct problems, and conduct disorder. MAIN OUTCOME MEASURES: Costs in 1998 prices for public services (excluding private, voluntary agency, indirect, and personal costs) used over and above basic universal provision. RESULTS: By age 28, costs for individuals with conduct disorder were 10.0 times higher than for those with no problems (95% confidence interval of bootstrap ratio 3.6 to 20.9) and 3.5 times higher than for those with conduct problems (1.7 to 6.2). Mean individual total costs were 70 019 pounds sterling for the conduct disorder group (bootstrap mean difference from no problem group 62 pound sterling; 898 pound sterling 22 692 pound sterling to 117 pound sterling) and 24 324 pound sterling (16 707 pound sterling; 6594 pound sterling to 28 149 pound sterling) for the conduct problem group, compared with 7423 pound sterling for the no problem group. In all groups crime incurred the greatest cost, followed by extra educational provision, foster and residential care, and state benefits; health costs were smaller. Parental social class had a relatively small effect on antisocial behaviour, and although substantial independent contributions came from being male, having a low reading age, and attending more than two primary schools, conduct disorder still predicted the greatest cost. CONCLUSIONS: Antisocial behaviour in childhood is a major predictor of how much an individual will cost society. The cost is large and falls on many agencies, yet few agencies contribute to prevention, which could be cost effective. PMID- 11473908 TI - Multicentre controlled trial of parenting groups for childhood antisocial behaviour in clinical practice. AB - OBJECTIVE: To see whether a behaviourally based group parenting programme, delivered in regular clinical practice, is an effective treatment for antisocial behaviour in children. DESIGN: Controlled trial with permuted block design with allocation by date of referral. SETTING: Four local child and adolescent mental health services. PARTICIPANTS: 141 children aged 3-8 years referred with antisocial behaviour and allocated to parenting groups (90) or waiting list control (51). INTERVENTION: Webster-Stratton basic videotape programme administered to parents of six to eight children over 13-16 weeks. This programme emphasises engagement with parental emotions, rehearsal of behavioural strategies, and parental understanding of its scientific rationale. MAIN OUTCOME MEASURES: Semistructured parent interview and questionnaires about antisocial behaviour in children administered 5-7 months after entering trial; direct observation of parent-child interaction. RESULTS: Referred children were highly antisocial (above the 97th centile on interview measure). Children in the intervention group showed a large reduction in antisocial behaviour; those in the waiting list group did not change (effect size between groups 1.06 SD (95% confidence interval 0.71 to 1.41), P<0.001). Parents in the intervention group increased the proportion of praise to ineffective commands they gave their children threefold, while control parents reduced it by a third (effect size between groups 0.76 (0.16 to 1.36), P=0.018). If the 31 children lost to follow up were included in an intention to treat analysis the effect size on antisocial behaviour was reduced by 16%. CONCLUSIONS: Parenting groups effectively reduce serious antisocial behaviour in children in real life conditions. Follow up is needed to see if the children's poor prognosis is improved and criminality prevented. PMID- 11473910 TI - Changing patterns of teenage pregnancy: population based study of small areas. AB - OBJECTIVES: To measure the impact of socioeconomic deprivation on rates of teenage pregnancy and the extent of local variation in pregnancy rates in Scotland, and to examine how both have changed over time. DESIGN: Population study using routine data from hospital records, aggregated for small areas. SUBJECTS: Female teenagers resident in Scotland who were treated for pregnancy in an NHS hospital in either 1981-5 (62 338 teenagers) or 1991-5 (48 514) and who were aged 13-19 at the time of conception. MAIN OUTCOME MEASURES: Pregnancy rates per 1000 in age group and the proportions of pregnancies resulting in a maternity (live birth or stillbirth) in teenagers aged 13-15, 16-17, and 18-19. RESULTS: From the 1980s to the 1990s pregnancy rates increased differentially according to levels of local deprivation, as measured by the Carstairs index. Among teenagers aged less than 18 the annual pregnancy rate increased in the most deprived areas (from 7.0 to 12.5 pregnancies per 1000 13-15 year olds and from 67.6 to 84.6 per 1000 16-17 year olds), but there was no change, on average, among teenagers in the most affluent areas (3.8 per 1000 13-15 year olds and 28.9 per 1000 16-17 year olds). Among 18-19 year olds the pregnancy rate decreased in the most affluent areas (from 60.0 to 46.3 per 1000) and increased in the most deprived areas (from 112.4 to 116.0 per 1000). The amount of local variation explained by deprivation more than doubled from the 1980s to the 1990s. The proportion of pregnancies resulting in a maternity was positively associated with level of deprivation, but the effect remained similar over time. CONCLUSION: From the 1980s to the 1990s the difference in rates of teenage pregnancy between more affluent and more deprived areas widened. This has implications for allocating resources to achieve government targets and points to important social processes behind the general increase in the number of teenage pregnancies in Scotland. PMID- 11473911 TI - "They're doing people a service"-qualitative study of smoking, smuggling, and social deprivation. AB - OBJECTIVES: To examine the behaviour and attitudes related to smoking and contraband tobacco products among smokers in two socially deprived areas. DESIGN: Cross sectional study with qualitative semistructured interviews, augmented by smokers' day grid. SETTING: Two areas of socioeconomic deprivation in Edinburgh. PARTICIPANTS: 50 male and 50 female smokers aged 25-40 years randomly selected from general practitioners' lists from two health centres, each located in an area of deprivation. RESULTS: Most smokers wanted to quit but felt unable to because of the importance of smoking in their daily routine and their addiction to nicotine. Strategies for maintaining consumption levels in the face of increasing cigarette prices and low income included purchasing contraband cigarettes and tobacco. Vendors were contacted through social networks, family, and friends as well as common knowledge of people and places, particularly pubs where contraband was available. Most users of contraband considered that smugglers were providing a valuable service. Purchasing contraband tobacco was viewed as rational in the face of material hardship. Many smokers criticised the government for its high tobacco taxation and the lack of local services to help them to stop smoking. CONCLUSIONS: Smokers in deprived areas perceive a lack of support to help them to stop smoking. Cigarette and tobacco smuggling is therefore viewed positively by low income smokers as a way of dealing with the increasing cost of cigarettes. Smokers in areas of deprivation may thus show little support for tackling smuggling until more action is taken to deal with the material and personal factors that make it difficult for them to quit. PMID- 11473912 TI - Mortality in children registered in the Finnish child welfare registry: population based study. PMID- 11473913 TI - Social environments and health: cross sectional national survey. PMID- 11473915 TI - The health of children in refuges for women victims of domestic violence: cross sectional descriptive survey. AB - OBJECTIVES: To describe the health and developmental status of children living in refugees for women victims of domestic violence and to investigate their access to primary healthcare services. DESIGN: Cross sectional survey. SETTING: Women's refugees in Cardiff. PARTICIPANTS: 148 resident children aged under 16 years and their mothers. MAIN OUTCOME MEASURES: Completeness of records on the child health system (register of all children that includes data on the child's health) for named health visitor, named general practitioner, and immunisation uptake; satisfactory completion of child health surveillance; Denver test results for developmental status; Rutter test scores for behavioural and emotional problems; reports of maternal concerns. RESULTS: 148/257 (58%) children living in refugee between April 1999 and January 2000 were assessed. Child health system data were incorrect (general practitioner and/or address) or unavailable for 85/148 (57%) children. Uptake of all assessments and immunisations was low. 13/68 (19%) children aged <5 years had delayed or questionable development on the Denver test, and 49/101 (49%) children aged 3-15 years had a Rutter score of >10 (indicating probable mental health problems). Concerns were expressed by mothers of 113/148 (76%) children. After leaving the refuge, 22 children were untraceable and 36 returned home to the perpetrator from whom the families had fled. CONCLUSIONS: The children had a high level of need, as well as poor access to services. Time spent in a refuge provides a window of opportunity to review health and developmental status. Specialist health visitors could facilitate and provide support, liaison, and follow up. PMID- 11473916 TI - Barriers to uptake of services for coronary heart disease: qualitative study. AB - OBJECTIVES: To identify factors within the South Yorkshire coalfields that influence use of health services by people with angina. DESIGN: Qualitative study using semistructured individual and group interviews. SETTING: General practice and community settings in Barnsley and Rotherham health authorities. PARTICIPANTS: 14 patients with stable angina and nine primary care staff had individual interviews plus five community groups and one group of general practitioners. MAIN OUTCOME MEASURES: Barriers to accessing health services. RESULTS: A complex web of factors was identified that prevented, delayed, or facilitated referral to secondary care. Delay, denial, and self management by patients meant that the full extent of symptoms often remained hidden from general practitioners, resulting in a delayed or missed referral. Barriers identified fell into six categories: structural, personal, social and cultural, past experience and expectations, diagnostic confusion, and knowledge and awareness. CONCLUSIONS: Many of the factors influencing referral operate before general practitioners become involved. Community development could be one way of tackling inequalities and promoting sustainable change. Structural changes are needed to improve access and increase the acceptability of general practice services. Primary care staff should be educated to detect underreporting of symptoms and promote appropriate referral. PMID- 11473918 TI - Trachoma. PMID- 11473920 TI - Equity in the new NHS: hard lessons from implementing a local healthcare policy on donepezil. PMID- 11473921 TI - Systematic reviews of evaluations of prognostic variables. PMID- 11473926 TI - Olfactory marker protein immunohistochemistry and the anterograde transport of horseradish peroxidase as indices of damage to the olfactory epithelium. AB - The present study compared the relative effectiveness of wheatgerm agglutinin- horseradish peroxidase (WGA--HRP) and olfactory marker protein (OMP) in detecting the presence of intact olfactory axons in glomeruli of the main olfactory bulb (MOB) in the rat. The olfactory epithelium was damaged by i.p. injections of the toxin 3-methyl indole and, after 5 or 6 days, the olfactory sac was injected with a 1% WGA--HRP solution. No anterograde labeling was observed in the dorsal and ventromedial quadrants of the MOB in the WGA--HRP material. However, in the same cases OMP immunostaining was observed throughout the MOB. In other rats the rostral olfactory epithelium was aspirated unilaterally and after 3, 11 and 16 days the olfactory sacs were injected with WGA--HRP and rats were perfused 1 day later. In these cases WGA--HRP reaction product was absent in the dorsolateral quadrant of the MOB on the aspirated side in all animals, but OMP immunostaining could be detected in the 4 and 12 day survival animals but not in the 17 day survival rat. These findings indicate that anterograde transport of WGA--HRP accurately reflects the presence of intact axons en route to the MOB. In contrast, OMP immunostaining persists in axon terminals severed from their parent cell body for at least 12 days and is a less useful marker of intact olfactory axons in experiments using short survival times. PMID- 11473927 TI - Source and cyclic release pattern of (Z)-7-dodecenyl acetate, the pre-ovulatory pheromone of the female Asian elephant. AB - Female Asian elephants (Elephas maximus) release a pre-ovulatory urinary pheromone, (Z)-7-dodecenyl acetate (Z7--12:Ac), to signal males of their readiness to mate. Z7--12:Ac is quantitatively elevated during the follicular stage of estrus, reaching maximum concentrations just prior to ovulation, as demonstrated by two complementary headspace techniques: (i) evacuated canister capture followed by cryogenic trapping; (ii) solid phase microextraction (SPME), used prior to gas chromatography/ mass spectrometry (GC/MS). These patterns were coincident with observed male behaviors and were consistent with biochemical and binding properties of the active ligand, including optimal binding pH. To release maximum amounts of Z7--12:Ac for quantitation, serum and urine samples from three mature female Asian elephants in their luteal and follicular stages of several estrous cycles were subjected to heat and pH changes and were then treated with protease prior to SPME-GC/MS analyses. When the post-luteal serum progesterone concentrations declined to baseline levels, Z7--12:Ac became detectable in the female urine. Throughout the follicular stage pheromone concentrations increased linearly with no apparent relationship to the two serum luteinizing hormone peaks. Pre-ovulatory urine also contained related compounds, including (Z)-7--12 dodecenol. The relative amount of this alcohol increased relative to acetate during long-term storage, with a proportional reduction in bioactivity. Z7--12:Ac was not detected in mucus samples from the urogenital tract. A potential precursor of Z7--12:Ac was identified in liver homogenates from female elephants in the follicular stage. PMID- 11473928 TI - fMRI activation in response to odorants orally delivered in aqueous solutions. AB - During food intake flavor perception results from simultaneous stimulation of the gustatory, olfactory and trigeminal systems. Olfactory stimulation occurs mainly through the retronasal pathway and the resulting perception is often interpreted as a taste perception, thus leading to the well-known sensory confusion between taste and olfaction. The present experiment was designed to study, with functional magnetic resonance imaging (fMRI), the cortical representation of olfactory perception in humans in response to retronasal stimulation by odorants delivered in aqueous solution. Psychophysical evaluation confirmed that the stimuli acted as pure olfactory stimuli through the retronasal pathway and did not present any taste component. Results showed activation in all brain regions previously described with neuroimaging techniques using olfactory stimulation with an odorized air flow. Piriform and orbitofrontal cortex were found activated as well as the hippocampal region, the amygdala, the insular lobe, the cingulate gyrus and the cerebellum. These results demonstrate the feasibility of efficiently stimulating the olfactory system in an fMRI scanner through the retronasal pathway with liquids delivered to the oral cavity. The presentation of olfactory stimuli in liquids to the mouth is a realistic model for the study of food-related flavor perception. This stimulation protocol furthermore allows presenting taste and olfactory stimuli separately or combined, thus allowing for direct comparisons between single modality representation, taste or olfaction, and representation of multi-modality mixtures. PMID- 11473929 TI - The oral sensation of carbonated water: cross-desensitization by capsaicin and potentiation by amiloride. AB - The oral sensation elicited by carbonated water is reduced by capsaicin and by blockers of carbonic anhydrase. We have investigated the temporal profile of this sensation and its cross-desensitization by capsaicin. We additionally tested if the sensation is influenced by amiloride. Following pretreatment of half of the dorsal tongue with 33 p.p.m. capsaicin, carbonated water was flowed over the tongue bilaterally for 5, 15 or 60 s. Subjects then performed a two-alternative forced choice test by indicating which side of the tongue had a stronger sensation and separately rated the sensory intensity on each side. Capsaicin significantly reduced the intensity of sensation elicited by carbonated water, consistent with cross-desensitization. This effect was weaker at 60 s because of a significant decline (desensitization) in ratings of the intensity of carbonated water on both sides of the tongue. Pretreatment with amiloride resulted in a small but significant increase in the intensity of the sensation elicited by the 15 s carbonated water stimulus, suggesting an amiloride-sensitive transduction mechanism. PMID- 11473930 TI - Histochemical identification of carbohydrate moieties in the accessory olfactory bulb of the mouse using a panel of lectins. AB - Lectin binding patterns in the olfactory bulb of the mouse were investigated using 12 biotinylated lectins. Three, with specificities for galactose, N acetylgalactosamine and L-fucose, stained only the nervous and glomerular layers of the accessory olfactory bulb; four, with specificities for galactose or N acetylglucosamine, stained these layers in both the accessory and the main olfactory bulbs; three, with specificities for N-acetylgalactosamine or L-fucose, effected general staining with little contrast between the background and the accessory olfactory bulb or other structures; the remaining two, both of them specific for mannose, stained no part of the tissue studied. In the nervous and glomerular layers of the accessory olfactory bulb six lectins stained the anterior and posterior halves with different intensities and two of these six similarly differentiated between rostral and caudal regions of the posterior half. We conclude that: (i) three lectins binding to different monosaccharides are specific stains for the vomeronasal system when used in this area of the mouse brain; (ii) it may be appropriate to distinguish three parts in the mouse accessory olfactory bulb, instead of the hitherto generally accepted two. PMID- 11473931 TI - Biogenic amines modulate olfactory receptor neurons firing activity in Mamestra brassicae. AB - The modulatory effects of the biogenic amines octopamine and serotonin on pheromonal receptor neurons of Mamestra brassicae were investigated. The responses to sex pheromone components of two cells types (A and B) in single male long sensilla trichodea were monitored. Cell types A and B do not respond to the same compound. The response of type A to a pulse of the major sex pheromone component increased 5 min after octopamine injection. Responses of type B to other odorants increased after 30 min. In the absence of any pheromone stimulation the background firing activity of type A increased following octopamine injection. This background activity was used to evaluate the kinetics of octopamine and other biogenic amine effects on olfactory receptor neurons. Octopamine increased this background activity in a concentration- and time dependent manner. Clonidine, an octopamine agonist, was shown to be more powerful in increasing the background activity of olfactory receptor neurons. The effects of octopamine and clonidine were hypothesized to arise from specific receptor activation as chlorpromazine (an octopamine antagonist) was shown to block the effect of octopamine. Serotonin, a known neuromodulator in most animal species, induced a reversible inhibition of spike firing. Altogether, these results indicate that biogenic amines can modulate the sensitivity of olfactory receptor neurons of moths either directly or by an action on adaptation. PMID- 11473932 TI - A simple and reliable test of olfactory learning and memory in mice. AB - The present paper describes a quick and efficient method for assessing olfactory discrimination learning in mice. In training mice received trials in which one odor (CS+) was paired with sugar and another odor (CS-) was paired with no sugar. When the mice were subsequently placed in a chamber with CS+ odor at one end and CS- odor at the other, they spent more time digging in CS+ than in CS- odor. In Experiment 2 mice trained with this procedure and tested after 60 days also spent more time digging in CS+ than CS- in the test phase, indicating that this olfactory discrimination task is effective for assessing long-term memory. In addition to the outbred strain of CD1 mice used in Experiments 1 and 2, C57Bl/6NCr/BR and DBA/2NCr/BR mice used in Experiment 3 also acquired this learned odor discrimination. Moreover, Experiment 4 showed that DBA animals were capable of acquiring this odor discrimination after receiving only two training trials (one exposure each to CS+ and CS-) per day for 4 days. PMID- 11473933 TI - Identification of a third rat odorant-binding protein (OBP3). AB - From a rat olfactory epithelium cDNA library clones encoding a lipocalin were isolated with sequence identity to the previously described salivary-specific alpha-2u globulin and the N-terminal region of mouse odorant-binding proteins OBP III and OBP-IV. In situ hybridization showed strong expression in nasal glands displaying a pattern equivalent to rat OBP1. Heterologously expressed protein was evaluated for its binding properties using spectroscopic approaches. The recombinant protein interacted with two fluorescent probes, 1-aminoanthracene (1 AMA) and 1,1'-bis(4-anilino-5-naphthalene)-sulfonic acid. 1-AMA binding was competed by several odorants with high affinity. The thermodynamic parameters of the protein-odorant interaction were determined using isothermal titration calorimetry. Due to its nasal expression and odorant-binding characteristics this protein was designated OBP3. PMID- 11473934 TI - Perception of trigeminal chemosensory qualities in the elderly. AB - One hundred healthy elderly subjects (65-88 years) were tested for their ability to: (i) assign verbal labels from a list of trigeminal type descriptors to six odorants known to have a strong trigeminal component; (ii) discriminate between intensity-matched pairs of these odorants in an odd-ball paradigm. Their performance was compared with that of 100 young controls (23--36 years). Young controls judged menthol and cineole as distinctly cool and fresh, acetic cid as pungent and sour and acetone as pungent, but showed no clear descriptive profile for ethanol and propanol. The descriptive profiles given by the elderly subjects correlated significantly with those given by the young controls for all six odorants and thus indicate a high degree of conformity in trigeminal perception of chemosensory qualities between the two age groups. In the odd-ball test the young controls correctly discriminated an average of 8.0 of 9 stimulus pairs presented, with most mistakes occurring in response to pairs with a similar trigeminal profile. With an average of 6.4 of 9 items correct, the discrimination performance of the elderly subjects was significantly poorer than that of the young controls but nevertheless significantly above chance at the group level with all 9 stimulus pairs. These results suggest that the nasal trigeminal system may experience some degree of age-related impairment but still contributes considerably to the perception and discrimination of chemosensory qualities in the elderly. PMID- 11473937 TI - Recommendations on the management of pulmonary hypertension in clinical practice. PMID- 11473938 TI - Publication ethics and human reproduction. PMID- 11473939 TI - Double publication in Human Reproduction Volume 15, Supplement 1, 2000. PMID- 11473940 TI - Prognostic value of Y deletion analysis. The role of current methods. AB - Y chromosome microdeletions represent the most frequent genetic alteration in azoospermic and severely oligozoospermic men, and screening for microdeletions in AZFa, b and c are routinely performed in the major andrology and infertility centres. Since patients with Y microdeletions often require intracytoplasmic sperm injection (ICSI), the question of whether the type of the microdeletion present could have prognostic value for the presence of spermatozoa in the ejaculate or in the testes [by testicular sperm extraction (TESE)] is an interesting one. The review of the literature on this topic showed that there is still no clear genotype--phenotype relationship, i.e. similar testicular alterations may be caused by different types of microdeletions, and apparently identical microdeletions may be associated with diverse tubular damage. Even in azoospermic men, the localization of the microdeletion cannot be used as a valid prognostic parameter before TESE--ICSI to identify patients with spermatozoa in their testes. The only finding with absolute negative prognostic value is the presence of complete AZFa--c deletions, which are invariably associated with an absence of spermatozoa. Microdeletions in AZFa or AZFb seem to have promising prognostic value, but more data and gene-specific deletions have to be provided to draw clear conclusions. The absence of a clear genotype--phenotype relationship, and therefore of a prognostic value of Y deletion analysis, is probably due to the current methods used for the screening of the microdeletions. In fact, to date most centres do not use gene-specific markers but instead use anonymous primers that contribute little information to the pathogenic role of the microdeletions. PMID- 11473941 TI - Peritoneal closure--to close or not to close. AB - Peritoneal closure is a controversial issue among obstetricians and gynaecologists. This article reappraises the issue of peritoneal closure. We conducted a thorough literature search using Medline, Pubmed and Embase as well as a hand-search for all references quoted in the relevant papers. The routine non-closure of the peritoneum reduces operation time by an average of 6 min. Most studies showed no difference in the other outcome measures including infection/febrile episodes, analgesic/anaesthetics requirement, bowel function restoration, post-operative stay and adhesion formation. There are insufficient data concerning adhesion formation. In conclusion, apart from a slightly shorter operation time associated with non-closure of the peritoneum, many studies showed no difference in short-term morbidity in the closure and the non-closure group. More studies are needed to examine the long-term morbidity associated with the closure or the non-closure of the peritoneum. PMID- 11473942 TI - Disposition of sperm donors with resultant abnormal pregnancies. AB - We wished to determine how clinicians manage sperm donors whose offspring have chromosomal or structural abnormalities. A directed, multiple-choice survey was given to reproductive endocrinologists and obstetrical geneticists to assess management of sperm donors whose offspring have chromosomal or structural abnormalities. The questionnaire was completed by 66 reproductive endocrinologists and obstetrical geneticists. Abnormalities and the most common inheritance modes included: Trisomy 21 (aneuploidy, maternal origin), Turner syndrome (aneuploidy, paternal origin), cleft lip/palate (multifactorial), VATER sequence (vertebral defects, imperforate anus, tracheo-esophageal fistula, radial and renal dysplasia, sporadic inheritance), and Hurler syndrome (autosomal recessive). Response choices were: (i) remove donor from programme, (ii) inform potential recipients of prior pregnancy outcomes and continue to use donor, or (iii) further study donor to assess karyotype/mutations. Inheritance mode appeared to influence decisions to remove donors from sperm banks; however, no clear consensus was noted. Guidelines exist for screening potential gamete donors, but not for managing donors whose offspring has a chromosomal or structural abnormality. Guidelines must be developed to manage sperm donors with untoward pregnancy outcomes. PMID- 11473943 TI - Endometrial angiogenesis throughout the human menstrual cycle. AB - BACKGROUND: The timing and mechanisms of new blood vessel formation in the endometrium during the menstrual cycle are still largely unknown. In the present study we used the chick embryo chorioallantoic membrane (CAM) as an in-vivo assay for angiogenesis to assess the angiogenic potential of endometrium obtained at different stages of the menstrual cycle. METHODS: Endometrial fragments were explanted onto the CAM and, after 4 days of incubation, slides of the treated area were taken in ovo through a microscope for computerized image analysis. The vascular density index (VDI), a stereological estimate of vessel number and length, was obtained by counting the intersections of vessels with five concentric circles of a circular grid superimposed on the computerized image. RESULTS: We demonstrated that human endometrium has angiogenic potential throughout the menstrual cycle. Furthermore, there was a significant difference in angiogenic response between the stages of the menstrual cycle (P = 0.01). The VDIs of the early proliferative, early and late secretory stage were significantly higher than the VDI of the late proliferative phase. CONCLUSIONS: Elongation of existing vessels during the early proliferative phase as well as growth and coiling of the spiral vessels during the secretory phase may demand far higher angiogenic activity than outgrowth and maintenance of vessels during the late proliferative phase. PMID- 11473944 TI - Reversible suppression of menstruation with progesterone antagonists in rhesus macaques. AB - BACKGROUND: A reliable means of menstrual suppression would greatly improve the quality of life for women. Information is lacking on the direct endometrial effects and appropriate dosages of new antiprogestins that may be useful for this purpose. METHODS: The current work evaluated three different systems in macaque monkeys. First, the range of doses of two relatively new antiprogestins, ZK 137 316 and ZK 230 211, that would block progesterone action directly on the endometrium in artificially cycled, spayed rhesus macaques; second, the direct endometrial effects of ZK 230 211, a type III antiprogestin; and third, investigation of whether endometrial-suppressive doses administered chronically to intact, cycling monkeys could be used for reversible, menstrual suppression. RESULTS: The results in naturally cycling animals showed that ZK 137 316 blocked menstruation in all animals, but doses of 0.05 mg/kg blocked ovulation in 55.5% of animals and doses of 0.1 mg/kg blocked ovulation in 66.6% of the animals. However, all doses of ZK 230 211 that blocked menstruation also blocked ovulation. All progesterone antagonist (PA)-treated animals, regardless of dose, maintained normal follicular phase concentrations of oestradiol and returned to normal menstrual cyclicity within 15--41 days post-treatment. Therefore ZK 137 316, depending on dose, can allow ovulation but block menstruation, while ZK 230 211, a much more potent PA, blocks both ovulation and menstruation at all effective doses. Both PAs block unopposed oestrogenic action on the endometrium through their antiproliferative effects. CONCLUSIONS: Reversible amenorrhoea can be achieved with these two PAs, and they can protect the endometrium from the effects of unopposed oestrogen whether or not ovulation is blocked. Chronic, low dose PA treatment may provide a new option for women who wish to suppress their menstrual periods. PMID- 11473945 TI - Live human germ cells in the context of their spermatogenic stages. AB - BACKGROUND: Various types of live, dispersed, human testicular cells in vitro were previously compared with the morphologic characteristics of human spermatogenic germ cells in situ within seminiferous tubules. The current study extends those observations by placing live human germ cells in the context of their developmental steps and stages of the spermatogenic cycle. METHODS: Live human testicular tissue was obtained from an organ-donating, brain-dead person. A cell suspension was obtained by enzymatic digestion, and dispersed cells were observed live with Nomarski optics. Testes from 10 men were obtained at autopsy within ten hours of death, fixed in glutaraldehyde, further fixed in osmium, embedded in Epon, sectioned at 20 microm, and observed unstained by Nomarski optics. RESULTS: In both live and fixed preparations, Sertoli cells have oval to pear-shaped nuclei with indented nuclear envelopes and large nucleoli, which makes their appearance distinctly different from germ cells. For germ cells, size, shape, and chromatic pattern of nuclei, the presence of meiotic metaphase figures, acrosomic vesicles/structures, tails, and/or mitochondria in the middle piece are characteristically seen in live dispersed cells and those in the fixed seminiferous tubules. These lead to identification of live germ cells in man and placement of each in the context of their developmental steps of spermatogenesis at corresponding stages of the spermatogenic cycle. CONCLUSIONS: This comparative approach allows verification of the identity of individual germ cells seen in vitro and provides a checklist of distinguishing characteristics of live human germ cells to be used in clinical procedures or by scientists interested in studying live cells at known steps in spermatogenic development characteristic of germ cells in specific stages of the spermatogenic cycle. PMID- 11473946 TI - PAF receptor and PAF acetylhydrolase expression in the endosalpinx of the human Fallopian tube: possible role of embryo-derived PAF in the control of embryo transport to the uterus. AB - BACKGROUND: Prostaglandin-E(2) and platelet-activating factor (PAF) are embryonic derived signals that time embryo passage into the uterus in the mare and hamster respectively. PAF-like activity is detectable in the spent media of preimplantation human embryos and it has been suggested that PAF may be the embryonic signal that controls embryo transport to the uterus in our species. The actions of PAF are regulated at the level of its synthesis and degradation as well as the expression of a specific cell surface receptor (PAFr). The enzyme PAF acetylhydrolase (PAF-AH) degrades PAF. This study was undertaken to examine whether or not PAFr and PAF-AH are expressed in the human Fallopian tube and to identify the cell types in which they are expressed. METHODS: The presence of PAFr mRNA in tissue extracts was investigated using reverse transcription polymerase chain reaction. We amplified the predicted amplicon for PAFr mRNA from RNA samples extracted from Fallopian tubes. The expression of PAF-AH was detected by Western blot and the localization of PAFr and PAF-AH proteins was detected by immunohistochemistry. RESULTS: Utilizing antibodies against PAFr and PAF-AH, co localization of the two proteins in the epithelium and stromal cells were demonstrated. CONCLUSIONS: These observations show that the human Fallopian tube expresses PAFr and PAF-AH at a location compatible with the proposed paracrine role of early embryo-derived PAF. PMID- 11473947 TI - Perinatal outcome of pregnancy after GnRH antagonist (ganirelix) treatment during ovarian stimulation for conventional IVF or ICSI: a preliminary report. AB - BACKGROUND: Gonadotrophin-releasing hormone (GnRH) antagonists have been proven safe and effective, with no adverse effects on offspring in animal studies. Careful study of pregnancy outcome in humans is mandatory. METHODS AND RESULTS: This preliminary report includes follow-up data of patients treated with the GnRH antagonist, ganirelix, during ovarian stimulation for IVF or ICSI. In total, 333 patients were randomized in a multicentre, double-blind, dose-finding study of ganirelix, at six different doses ranging from 0.0625 to 2 mg. In total, 68 vital intrauterine pregnancies were established that resulted in the birth of 46 singletons, 12 twins and one triplet. Follow-up of the 67 pregnant patients (one subject was lost to follow-up) revealed six miscarriages (9%). Of the 61 subjects with an ongoing pregnancy, two with a singleton pregnancy did not give birth to a live-born infant (one spontaneous abortion in week 19, and one intrauterine death in week 27). The mean gestational age was 39.4 weeks for singleton pregnancies, and 36.6 weeks for multiple pregnancies. In total, 73 infants (33 boys, 40 girls) were born. A birth weight <2500 g was reported for 8.7% and 54.2% of the infants resulting from singleton and twins delivery respectively. One major congenital malformation was diagnosed; a boy with Beckwith-Wiedemann syndrome (exomphalos and macroglossia). Seven minor malformations were reported among five infants. CONCLUSIONS: In this first follow-up study, the incidence of adverse obstetrical and neonatal outcome was comparable with reported incidences for IVF-embryo transfer pregnancies. PMID- 11473948 TI - First human exposure to FSH-CTP in hypogonadotrophic hypogonadal males. AB - BACKGROUND: This is the first report of human exposure to the novel compound follicle stimulating hormone (FSH)-C-terminal peptide (CTP) 'FSH-CTP' (Org 36286), a long-acting recombinant FSH like substance, consisting of the alpha subunit of human FSH and a hybrid beta-subunit. The latter is composed of the beta-subunit of human FSH and the C-terminus part (CTP) of the beta-subunit of human chorionic gonadotrophin (HCG). METHODS: In this phase I, non-blind, multi centre study, 13 hypogonadotrophic hypogonadal male subjects were enrolled to test the safety of FSH-CTP in terms of antibody formation in humans. Furthermore, the pharmacokinetic profile of this new compound was determined. Subjects were injected four times with 15 microg FSH-CTP with an interval of approximately 4 weeks between each injection. RESULTS: No drug related (serious) adverse events occurred. No antibodies against FSH-CTP or chinese hamster ovary (CHO)-cell derived proteins were detected and measurement of local tolerance demonstrated that s.c. administration of FSH-CTP is well tolerated and no increase in intensity of injection-site responses was observed after repeated exposure to FSH CTP. After the first and third injection, FSH-CTP serum concentrations were determined. Overall mean (+/- SD) C(max) was 0.426 (+/- 0.116) ng/ml, mean t(1/2) and AUC(0-infinity) were 94.7 (+/- 26.2) h and 81.5 (+/- 18.8) ng.h/ml respectively. Compared with recFSH (Puregon), the half life of FSH-CTP was increased 2-3 times. Following the first and third injection a clear rise in serum inhibin-B concentrations were observed. CONCLUSIONS: The use of FSH-CTP is safe and does not lead to detectable formation of antibodies. Furthermore, the pharmacokinetic and dynamic profile of FSH-CTP may lead to the development of new, more convenient regimens for the treatment of male and female infertility. PMID- 11473949 TI - Resumption of ovarian function during lactational amenorrhoea in breastfeeding women with polycystic ovarian syndrome: metabolic aspects. AB - BACKGROUND: Polycystic ovarian syndrome (PCOS) is a common endocrine-metabolic disorder in women, a high percentage of whom exhibit peripheral insulin resistance. After delivery, in normal women, lactation imposes a metabolic adaptation, the impact of which on the insulin resistance of PCOS patients is not known. The aim of this study was to evaluate the effect of lactation on insulin resistance, glucose and insulin metabolism, and sex hormone-binding globulin (SHBG) and insulin-like growth factor binding protein-1 (IGFBP)-1 concentrations in fully breast-feeding normal and PCOS women during the postpartum period (lactational amenorrhoea) and also after weaning. METHODS: Twelve lactating PCOS (LPCOS) women and six normal lactating (NL) women of similar age and body mass index (BMI) were selected for the study. At the 4th and the 8th week postpartum (pp), and 8 weeks after weaning, a 2 h, 75 g oral glucose tolerance test (oGGT) was performed, followed by an insulin tolerance test 2 days later. For the oGGT, glucose and insulin were measured in each sample and SHBG and IGFBP-1 were determined in the fasting sample. RESULTS: During lactation, fasting insulin levels were similar in both groups. In LPCOS women 2 h insulin concentrations were significantly higher, and SHBG and IGFBP-1 concentrations were significantly lower, than those observed in NL women. In both groups, insulin sensitivity evaluated by the insulin tolerance test was not modified. After weaning, in LPCOS women, SHBG and IGFBP-1 concentrations remained lower and insulin concentrations remained higher than those observed in NL women ( P < 0.05 ). CONCLUSIONS: In PCOS women, insulin resistance is not modified during lactation. Lactation has a transitory beneficial effect on insulin levels and biological markers of insulin resistance. PMID- 11473950 TI - Resumption of ovarian function during lactational amenorrhoea in breastfeeding women with polycystic ovarian syndrome: endocrine aspects. AB - BACKGROUND: The aim of this study was to evaluate the changes in gonadotrophin concentrations and the dynamics of the episodic fluctuations of circulating LH during night-time, in fully breastfeeding normal women and in those with polycystic ovarian syndrome (PCOS) during lactational amenorrhoea and after weaning, in order to provide insights into the onset of this syndrome. Additionally, ovarian activity was evaluated by ultrasound examination and steroid concentrations. METHODS: Twelve lactating PCOS (LPCOS) women and six normal lactating (NL) women of similar age were selected. On the 4th and 8th week postpartum (PP) and eight weeks after weaning, blood samples were collected every 10 min (10.00--20.00h). Gonadotrophin concentrations were determined in all samples. Steroid hormones were measured in one fasting sample and ovarian morphology was assessed by ultrasound. RESULTS: On the 8th week PP, LH pulse frequency was higher and FSH concentrations were lower in LPCOS women compared with NL women, and steroid hormone concentrations remained low, except for androstenedione which was higher in LPCOS patients. After weaning, similar differences were observed between both groups. PCOS patients also showed enlarged ovaries with a PCOS pattern in the three study periods. CONCLUSIONS: The enlarged ovaries associated with higher androstenedione concentrations suggest that PCOS is a primary ovarian defect, making it difficult to establish if the abnormal LH pattern observed in these women is primary or secondary to the ovarian dysfunction. PMID- 11473951 TI - Changes in the biological:immunological ratio of basal and GnRH-releasable FSH during the follicular, pre-ovulatory and luteal phases of the human menstrual cycle. AB - BACKGROUND: Significant changes in charge isoform distribution of serum FSH occur throughout the human menstrual cycle. In the present study, we analysed the impact of the changing endocrine milieu characteristic of the menstrual cycle on the capability of basal and gonadotrophin-releasing hormone (GnRH)-releasable FSH to trigger intracellular signal transduction via the human FSH receptor. METHODS: Seven normal women underwent blood sampling every 10 min for 10 h during the early follicular phase (FP), pre-ovulatory phase (PO) and mid- to late luteal phase (LP) of the menstrual cycle. Serum from successive samples collected across 2 h intervals containing FSH released under baseline and exogenous GnRH stimulated conditions was tested for bioactivity employing a homologous in-vitro assay. RESULTS: The biological to immunological (B:I) ratio of basal and GnRH releasable FSH was significantly (P < 0.05 ) higher at LP (range, 0.83 +/- 0.07 to 1.35 +/- 0.30) than during the FP (0.43 +/- 0.02 to 0.65 +/- 0.04) and PO (0.49 +/- 0.05 to 0.62 +/- 0.06). In all phases, the B:I FSH ratio in baseline samples was similar to those exhibited by samples collected after 10 and 90 microg GnRH administration. CONCLUSIONS: The selective increase in the capability of the admixture of FSH isoforms circulating during the LP to activate the FSH receptor, apparently represents an additional mechanism through which the anterior pituitary may regulate the maturation of those follicles destined to ovulate during the coming cycle. PMID- 11473952 TI - Prediction of the potentially fertile period by urinary hormone measurements using a new home-use monitor: comparison with laboratory hormone analyses. AB - BACKGROUND: The study compared a new urinary hormone monitoring system, Clearview Primera Fertility Monitor (CPFM), with laboratory hormone analyses in the prediction of the potentially fertile period. METHODS: Thirty healthy female volunteers provided blood and early morning urine samples for one cycle. Serum oestradiol, progesterone and luteinizing hormone (LH), and urinary LH and oestrone-3-glucuronide (E3G) were measured. The fertility status of volunteers; Low, High or Peak, was collected from monitors and compared with the hormone measurements. RESULTS: There was agreement between the first day of peak fertility and the urinary LH peak day in 65.6% of cycles and detection 1 or 2 days before the urinary LH peak day in 24.1 and 6.9% of cycles respectively. In 58.6% of cycles the system detected up to 5 days of increased fertility prior to the urinary LH peak day. Warning days of the urinary LH peak were similarly determined using defined thresholds of E3G and oestradiol providing up to 5 days warning in 82.8 and 96.6% of cycles respectively. CONCLUSIONS: The system can provide couples attempting to conceive with information about the potentially fertile days in the cycle in order that they may time intercourse. It also has potential for use in evaluation and treatment of infertile couples. PMID- 11473953 TI - Effects of metformin on ovulation rate, hormonal and metabolic profiles in women with clomiphene-resistant polycystic ovaries: a randomized, double-blinded placebo-controlled trial. AB - BACKGROUND: Metformin, an insulin-sensitizing agent, has been used successfully as the first-line drug to induce ovulation in women with polycystic ovary syndrome. There are, however, very few studies evaluating metformin treatment in women with clomiphene citrate (CC)-resistant polycystic ovaries (PCO). METHODS: Twenty infertile Chinese women aged <40 years, who had ultrasound features of PCO and remained anovulatory on CC, were randomized by computer using the sealed envelope method to receive placebo or metformin 500 mg three times a day for 3 months. Hormonal and metabolic profiles were determined before the therapy and were repeated after 3 months for women who failed to become pregnant within this period. Clomiphene was then added for one cycle to those women who did not ovulate after taking placebo or metformin alone. RESULTS: The median ovulation rate in the placebo group was 0% (range: 0--50%) after placebo only and 6.9% (range: 0--50%) after placebo and CC, whereas the corresponding rates in the metformin group were 0% (range: 0--22%) and 0% (range: 0--22%) respectively. There was no improvement in the ovulation rate despite a significant reduction of body mass index, serum testosterone and fasting leptin concentrations in the metformin group. CONCLUSIONS: Metformin treatment may result in successful ovulation only in certain subgroups of these women. PMID- 11473954 TI - Comparison between 1 year oral and transdermal oestradiol and sequential norethisterone acetate on circulating concentrations of leptin in postmenopausal women. AB - BACKGROUND: Oral and transdermal postmenopausal hormone replacement therapy (HRT) affects lipid and glucose metabolism differently, which is of significance in the release of leptin by adipocytes. Moreover, oestrogen and progesterone can stimulate leptin secretion in women of reproductive age. Therefore, we compared the effects of oral and transdermal oestrogen plus progestin regimen on plasma leptin in 38 healthy postmenopausal women with normal body mass index (BMI), who wished to use HRT to control incapacitating climacteric symptoms. METHODS: The women were randomized to treatment with oral HRT (2 mg oestradiol on days 1--12, 2 mg oestradiol plus 1 mg norethisterone acetate (NETA) on days 13--22, and 1 mg oestradiol on days 23--28, n = 19), or with transdermal HRT (50 microg/day of oestradiol on days 1--13, and 50 microg oestradiol plus 250 microg/day NETA on days 14--28, n = 19) for 1 year. Plasma samples were collected before and at oestradiol + NETA phase after 2, 6 and 12 months treatment and were assayed for leptin. RESULTS: The baseline leptin, ranging from 3.3 to 34.9 microg/l, was significantly associated with BMI (r = 0.78, P < 0.0001 ), but showed no difference between women in oral HRT (geometric mean 13.9 microg/l, 95% confidence interval (CI) 10.1--17.6 microg/l) or transdermal HRT group (geometric mean 12.0 microg/l, 95% CI 9.7--14.3 microg/l). Neither oral nor transdermal oestradiol + NETA caused any significant changes in plasma leptin (or BMI) after 2, 6, or 12 months of treatment. CONCLUSION: Leptin is an unsuitable factor to detect oestradiol + NETA-induced metabolic changes in postmenopausal women. PMID- 11473955 TI - Suppression of LH during ovarian stimulation: analysing threshold values and effects on ovarian response and the outcome of assisted reproduction in down regulated women stimulated with recombinant FSH. AB - BACKGROUND: It has been recently suggested that gonadotrophin-releasing hormone agonist down-regulation in some normogonadotrophic women may result in profound suppression of LH concentrations, impairing adequate oestradiol synthesis and IVF and pregnancy outcome. The aims of this study, where receiver-operating characteristic (ROC) analysis was used, were: (i) to assess the usefulness of serum LH measurement on stimulation day 7 (S7) as a predictor of ovarian response, IVF outcome, implantation, and the outcome of pregnancy in patients treated with recombinant FSH under pituitary suppression; and (ii) to define the best threshold value, if any, to discriminate between women with 'low' or 'normal' LH concentrations. METHODS: A total of 144 infertile women undergoing IVF/intracytoplasmic sperm injection (ICSI) treatment were included. Seventy-two consecutive patients having a positive pregnancy test (including 58 ongoing pregnancies and 14 early pregnancy losses) were initially selected. As a control non-pregnant group, the next non-conception IVF/ICSI cycle after each conceptual cycle in our assisted reproduction programme was used. RESULTS: The median and range of LH values in non-conception cycles, conception cycles, ongoing pregnancies, and early pregnancy losses, clearly overlapped. ROC analysis showed that serum LH concentration on S7 was unable to discriminate between conception and non-conception cycles (AUC(ROC) = 0.52; 95% CI: 0.44 to 0.61) or ongoing pregnancy versus early pregnancy loss groups (AUC(ROC) = 0.59; 95% CI: 0.46 to 0.70). To assess further the potential impact of suppressed concentrations of circulating LH during ovarian stimulation on the outcome of IVF/ICSI treatment, the three threshold values of mid-follicular serum LH proposed in the literature (<1, < or =0.7, <0.5 IU/l) to discriminate between women with 'low' or 'normal' LH were applied to our study population. No significant differences were found with respect to ovarian response, IVF/ICSI outcome, implantation, and the outcome of pregnancy between 'low' and 'normal' S7 LH women as defined by those threshold values. CONCLUSIONS: Our results do not support the need for additional exogenous LH supplementation in down-regulated women receiving a recombinant FSH-only preparation. PMID- 11473956 TI - Endothelial nitric oxide synthase gene polymorphism in women with idiopathic recurrent miscarriage. AB - BACKGROUND: Lack of endothelium-derived nitric oxide is associated with vasospasm and vascular infarction. We investigated the relationship between idiopathic recurrent miscarriage and a polymorphism of the gene encoding endothelial nitric oxide synthase (NOS3). METHOD: In a prospective case-control study, 105 women with idiopathic recurrent miscarriage and 91 healthy controls were investigated. We used the polymerase chain reaction to identify the different alleles of a 27 base pair tandem repeat polymorphism in intron 4 of the NOS3 gene. RESULTS: The wild type B allele was identified on 329 out of 392 chromosomes (frequency 0.84). The polymorphic A allele was present on 63 chromosomes (frequency 0.16). The genotype frequencies were as follows: 68% (B/B), 31% (A/B) and.5% (A/A). The distribution of genotype frequencies was significantly different between the study and control groups for allele A/B heterozygotes (NOS3(A/B)) (36.7 versus 23.8%, P = 0.03, OR 1.6, 95% CI 1.1--3.8). Only one individual was homozygous for the A allele (NOS3(A/A)). She was in the study group. Between women with primary and secondary recurrent miscarriages, no statistically significant difference between the distribution of NOS3(A/B) genotypes (28 versus 34%) was observed. CONCLUSIONS: These data support a role for the NOS3 gene as a genetic determinant of the risk of idiopathic recurrent miscarriage. PMID- 11473957 TI - Low-level sex chromosome mosaicism in female partners of couples undergoing ICSI therapy does not significantly affect treatment outcome. AB - BACKGROUND: There is an increased rate of chromosomal anomalies, in particular low-level sex chromosome mosaicism, in the female partners of couples undergoing intracytoplasmic sperm injection (ICSI). METHODS: Among 811 consecutive couples presenting for pre-ICSI chromosome analysis, chromosomal abnormalities were detected in 54 individuals, of which 26 were low-level sex chromosome mosaicism in the females. Attention was focused on the treatment course and outcome of ICSI in 20 couples with low-level sex chromosome mosaicism in the females actually embarking on ICSI treatment (group I, n = 38 ICSI treatment cycles). Applying a case-control design, each of the 20 couples was matched according to female age and source of spermatozoa to couples without a chromosomal abnormality in either of the partners (group II, n = 38 ICSI treatment cycles). RESULTS: No significant differences were found between the groups in ovarian response, fertilization rate and number of embryos transferred. Pregnancy rates, as well as implantation and abortion rates did not differ significantly between the groups. CONCLUSIONS: The data suggest that low-level sex chromosome mosaicism in females has no major effect on the course and outcome of ICSI. PMID- 11473958 TI - Mutation screening and CAG repeat length analysis of the androgen receptor gene in Klinefelter's syndrome patients with and without spermatogenesis. AB - BACKGROUND: Mutations of the androgen receptor (AR) gene give rise to a wide array of phenotypic abnormalities. A systematic analysis of the AR gene in patients with 47,XXY has not previously been performed. METHODS: Mutations of the AR gene and expansion of the CAG repeats in exon 1 of the AR gene were studied in 13 patients with Klinefelter's syndrome either with (n = 1) or without (n = 12) spermatogenesis. RESULTS: No abnormalities in the AR gene were detected by single strand conformational polymorphism analysis. The CAG lengths ranged from 17 to 27 (mean +/- SD 22.8 +/- 3.3, median 23) for Klinefelter patients or from 17 to 28 (mean +/- SD 23.2 +/- 2.6, median 23) for control subjects. X-inactivation analysis for the methylation status of the AR gene was performed in seven patients who were heterozygous for CAG repeats of different length, showing that the longer CAG repeat alleles underwent random but more frequent inactivation in five patients and skewed inactivation in two. CONCLUSIONS: An AR gene abnormality does not constitute an important factor for impaired spermatogenesis in patients with Klinefelter's syndrome. PMID- 11473959 TI - Oral contraceptive use in relation to age at menopause in the DOM cohort. AB - BACKGROUND: We investigated the hypothesis that long-term use of oral contraceptives (OCs), in particular high-dose OCs, could postpone age at menopause. METHODS: Data was used from 8701 women who participated in a breast cancer screening programme in Utrecht (DOM-3 cohort), and who did not use hormone replacement therapy (HRT) or OCs in the 4 years prior to their last menses. Data on OC-use, menopausal status, age at menopause, year of birth, parity, smoking behaviour, socio-economic status, body mass index and age at menarche was available. Use of high-dose OCs has been defined in this study as OC-use before 1972. The data was analysed by means of linear regression and Cox's proportional hazards analysis. Women still menstruating, women with surgical menopause and women lost to follow-up were censored at their last known date of menstruation. Endpoint was the natural menopause (n = 4589). RESULTS: The use of high-dose OCs advanced the onset of menopause by approximately 1.2 months for every year of OC use compared with no OC-use. High-dose OC-use for > or = 3 years, adjusted for confounding variables, increased the risk of earlier menopause compared with no OC-use (adjusted hazard ratio 1.12; 95% CI 1.03--1.21). The use of lower dose OCs did not increase the risk of earlier menopause (adjusted hazard ratio 1.00; 95% CI 0.91--1.09). CONCLUSIONS: These results are inconsistent with the hypothesis that long-term use of OCs could postpone the onset of menopause by inhibiting follicle depletion. Possible explanations are discussed. PMID- 11473960 TI - Utero-ovarian blood flow characteristics of pituitary desensitization. AB - BACKGROUND: Down-regulation in assisted reproduction treatment cycles is monitored by suppression of ovarian/pituitary hormones and/or measurement of endometrial thickness. METHODS: This prospective longitudinal study reports on utero-ovarian characteristics of pituitary desensitization. A total of 75 patients were recruited; 32 had IVF treatment, 20 frozen--thawed embryo transfer cycles and 23 patients were recipients of donated oocytes. All received early follicular-phase down-regulation and had colour flow Doppler velocimetry of the utero-ovarian arteries < or =3 days before the start of menses and after 21 days of gonadotrophin-releasing hormone (GnRH) analogue treatment. Ovarian volume, endometrial thickness, pituitary and ovarian hormone concentrations were recorded at each scan. RESULTS: Significant changes (P < 0.05) were noted in these and utero-ovarian vasculature during the down-regulation period, with good correlation between resistance index and oestradiol estimations. Neither the type of GnRH analogue nor age influenced the changes in utero-ovarian blood flow. Ovarian artery resistance index was the best Doppler predictor for pituitary suppression and a mean discriminatory cut-off value of 0.867 +/- 0.025 was found to have the highest specificity and positive predictive value. CONCLUSIONS: This study has, for the first time, defined cut-off values for satisfactory pituitary suppression with high positive predictive value and specificity in an early follicular phase long protocol of GnRH analogue down-regulation using colour flow Doppler. PMID- 11473961 TI - Continuous administration of gonadotrophin-releasing hormone agonist during the luteal phase in IVF. AB - BACKGROUND: It has been reported that ceasing the administration of gonadotrophin releasing hormone (GnRH) agonist causes a profound suppression of circulating serum gonadotrophins. A comparative prospective and randomized study was conducted to investigate the effect of continuous administration of GnRH agonist during the luteal phase in an ovarian stimulation programme for IVF. METHODS: GnRH agonist was administered intranasally from the midluteal phase of the previous cycle, and pure FSH administration started on cycle day 7. In the continuous-long protocol (cL) group (n = 161 ), GnRH agonist administration was continued until 14 days after oocyte retrieval. In the long protocol (L) group (n = 158 ), GnRH agonist was administered until the day before human chorionic gonadotrophin (HCG) administration. RESULTS: The implantation rate and live birth rate per unit of transferred embryos were significantly higher in the cL group than the L group (P < 0.05 ). Serum LH and FSH concentrations on the day of, and 1 day after, HCG administration were significantly lower in the L group than the cL group (P < 0.01 ). CONCLUSIONS: Continuation of GnRH agonist administration during the luteal phase might facilitate implantation, and prevent the profound suppression of serum gonadotrophins. PMID- 11473962 TI - A prospective, randomized comparison of two starting doses of recombinant FSH in combination with cetrorelix in women undergoing ovarian stimulation for IVF/ICSI. AB - BACKGROUND: A prospective randomized study was carried out in two centres to compare the number of oocytes retrieved after two different starting doses of recombinant human FSH (rhFSH) (Gonal-F) in women undergoing ovarian stimulation for IVF/intracytoplasmic sperm injection (ICSI) cycles using the multiple dose regimen of the gonadotrophin-releasing hormone (GnRH) antagonist cetrorelix (Cetrotide) to prevent induction of the premature LH surge. METHODS: Sixty women were randomized to receive rhFSH 150 IU ('low'), and 60 women to receive rhFSH 225 IU ('high') as the starting dose for the first 5 days of stimulation. From stimulation day 6 and onwards, including the day of human chorionic gonadotrophin (HCG) administration, the women received 0.25 mg of cetrorelix as a daily dose. The primary endpoint was the number of oocytes retrieved. RESULTS: The mean number (+/- SD) of oocytes was 9.1 +/- 4.4 and 11.0 +/- 4.6 in the 'low' and 'high' groups respectively (P = 0.024). The mean number of 75 IU ampoules of rhFSH was significantly lower in the 'low' group (23.0 +/- 6.3 versus 30.5 +/- 5.6, P < 0.0001). The ongoing pregnancy rate per started cycle and per embryo transfer were 25.9 and 28.8% versus 25.4 and 26.8% respectively in the 'low' and 'high' rhFSH groups (P = NS). CONCLUSIONS: When using a starting dose of 225 IU rhFSH combined with the multiple dose of 0.25 mg cetrorelix from stimulation day 6, significantly more oocytes were obtained than with a starting dose of 150 IU rhFSH. PMID- 11473963 TI - Perifollicular vascularity as a potential variable affecting outcome in stimulated intrauterine insemination treatment cycles: a study using transvaginal power Doppler. AB - BACKGROUND: The aim of the present study was to assess any potential relationship between perifollicular vascularity and outcome in an in-vivo environment following human chorionic gonadotrophin (HCG) administration. METHODS: A total of 182 unselected consecutive patients undergoing stimulated intrauterine insemination (IUI) cycles was recruited where the perifollicular vascularity of follicles > or =16 mm was studied using a subjective grading system and transvaginal power Doppler ultrasonography, 36 h after HCG administration. RESULTS: A total of 601 follicles was studied. The incidence of follicles showing high-grade perifollicular vascularity (3 and 4) was higher than those with low grade vascularity (1 and 2) (80 versus 20%). Treatment cycles were divided according to uniformity of vascularity grades of follicles > or =16 mm on the day of IUI [55% all high (3/4) grade; 33% mixed (1/2 and 3/4) and 12% all low (1/2) grade]. The mean age and duration of subfertility were significantly higher (P < 0.05), whereas the number of follicles > or =16 mm pre/post HCG, serum oestradiol and incidence of ultrashort gonadotrophin-releasing hormone (GnRH) agonist use were all significantly lower (P < 0.05) in treatment cycles with uniformly low follicular vascularity grades compared with mixed or uniformly high-grade cycles. However, on subjecting the data to multiple logistic regression analysis, the only independent variables that affected pregnancy rates appeared to be serum oestradiol (OR 1.28, 1.01--1.62) and high-grade follicular vascularity (OR 2.41, 1.08--5.40). CONCLUSION: These data would suggest that perifollicular vascularity has an important role to play in the outcome of IUI cycles, and that power Doppler has the potential to refine the management of assisted reproduction treatment cycles. PMID- 11473964 TI - Usefulness of transvaginal hydrolaparoscopy in investigating infertile women with Chlamydia trachomatis infection. AB - BACKGROUND: A new technique called transvaginal hydrolaparoscopy (THL) was recently developed for the exploration of the tubo-ovarian structures in infertile patients without obvious pelvic pathology. This study was performed to investigate the usefulness of THL to evaluate Chlamydia trachomatis tubal infertility. METHODS: Forty-one women with primary and secondary infertility participated in this study. Fourteen had past C. trachomatis infection. In 38 (92.7%) of the 41, access to the pouch of Douglas was obtained. In total, 71 (93.4%) out of 76 adnexa were clearly visualized. Thirty-seven patients were analysed and compared their tubal passages and peritubal adhesions using both hysterosalpingography (HSG) and THL. Twenty-four tubes from 14 patients with past C. trachomatis infection and 44 tubes from 23 patients without a history of C. trachomatis infection were compared. RESULTS: For the diagnosis of the tubal passage, there were no significant differences in the discrepancy rates between HSG and THL, in patients with and without past C. trachomatis infection. In 14 (58.3%) of the 24 tubes from patients with past C. trachomatis infection and in eight (18.2%) of the 44 tubes from patients without infection, peritubal adhesion was diagnosed only by THL. There was a significant difference in the discrepancy rates of the diagnosis of peritubal adhesion between HSG and THL in the two groups (P = 0.0007 ). CONCLUSIONS: These results suggest that C. trachomatis infection is highly associated with peritubal adhesion which is difficult to diagnose by HSG. Therefore, in C. trachomatis antibody-positive patients, exclusion of tubal pathology by THL or standard laparoscopy should be carried out to consider appropriate treatments. Although THL is not a substitute for laparoscopy, it can be proposed as a first line procedure in the early stages of the infertility investigation. PMID- 11473965 TI - Comparison of ovarian response in right and left ovaries in IVF patients. AB - BACKGROUND: Anatomical and cyclical physiological differences exist between right and left ovaries which may affect their function and response to ovulation induction. Although authors have compared right and left ovarian response during IVF for patients with a unilateral diseased or absent ovary, no study has examined the response of normal ovaries to gonadotrophin stimulation within the same patient. We wished to determine if there were any significant differences between right and left ovarian response in patients with healthy ovaries having standard IVF treatment. METHODS: We performed a prospective observational case- controlled study in 200 consecutive IVF patients. The main outcome measures were the number of oocytes retrieved, fertilization rates, grade of embryos produced, pregnancy rates and live birth rates. RESULTS: Comparison of right versus left ovary revealed: number of oocytes 4.9 versus 4.7, percentage fertilization 61.3 versus 62.5%, percentage of grade 1 embryos 81 versus 83%, chemical pregnancy rate 33 versus 47% and live birth rate 27 versus 32% (all not significant). CONCLUSIONS: We conclude that there are no statistical differences between right and left ovarian response in IVF patients with healthy ovaries. PMID- 11473966 TI - The development of an oocyte-containing follicle during gonadotrophin-releasing hormone agonist administration. AB - Administration of gonadotrophin-releasing hormone (GnRH) agonist in a 29 year old woman with infertility due to ovulatory dysfunction resulted in the development of several ovarian cysts. After human chorionic gonadotrophin (HCG) was injected, the cysts were aspirated and one mature oocyte was retrieved. Intracytoplasmic sperm injection (ICSI) was performed and the resulting embryo was transferred. A singleton pregnancy was obtained and a healthy baby was born at 36 weeks of gestation. Because GnRH agonist-derived cysts may contain oocytes, we suggest that when the growth of cysts is accompanied by high concentrations of oestradiol, the administration of HCG may be useful to achieve oocyte maturation and advance IVF treatment. PMID- 11473967 TI - Pregnancy and delivery after cryopreservation of zygotes produced by in-vitro matured oocytes retrieved from a woman with polycystic ovarian syndrome. AB - This case report describes the birth of a healthy infant after cryopreservation of zygotes produced by in-vitro matured oocytes retrieved from an anovulatory woman with polycystic ovarian syndrome (PCOS). To initiate the treatment cycle, the patient received intravaginal progesterone at night for 10 days to induce a withdrawal bleed. Oocyte retrieval was performed on day 11 following a withdrawal bleed. The patient was administered 10,000 IU of HCG subcutaneously 36 h prior to oocyte collection. A total of 63 immature oocytes were obtained; 10 were morphologically abnormal. Following incubation for 24--48 h in the maturation medium, TC-199 supplemented with 20% patient's own serum, 75 mIU/ml FSH and LH, 77.4% (41/53) of the oocytes were at the metaphase-II stage. Thirty-one (31/41, 75.6%) were fertilized using ICSI with her husband's spermatozoa, 15 fertilized oocytes were cultured for embryo transfer and 16 were frozen at the pronuclear stage. Pregnancy ensued following fresh embryo transfer. Unfortunately, the pregnancy was miscarried eight weeks later. However, the second frozen-thawed embryo transfer attempt resulted in a full-term pregnancy with delivery of a healthy male infant. PMID- 11473968 TI - Low rates of DNA fragmentation in selected motile human spermatozoa assessed by the TUNEL assay. AB - BACKGROUND: In this study we present the physiological changes observed in ejaculated spermatozoa of normospermic men after exposure to hydrogen peroxide (H(2)O(2)) or gamma irradiation. METHODS: Motility changes as well as membrane and DNA-damage were determined in spermatozoa after incubation with 25 micromol/l of H(2)O(2) during increasing intervals of time (0--60 min and after 24 h) or after irradiation of cells using alpha rays. Annexin V-binding in combination with propidium iodide was used for the assessment of membrane changes after each incubation time. TdT-mediated-dUTP nick-end labelling (TUNEL) was used to evaluate DNA damage. RESULTS: After 1 h incubation of the spermatozoa with H(2)O(2), almost all cells were positive for Annexin-V, while no significantly increase in TUNEL positivity was observed. TUNEL results were significantly higher 24 h after incubation with H(2)O(2) (10--16.3%, P = 0.03). In the control group (cumulus cells), an increase in the percentage of TUNEL positive cells was observed after 15 min of incubation with H(2)O(2) and showed a five-fold increase after 24 h (from 8.1-72.1%, P < 0.001). TUNEL positive cells after alpha irradiation increased with the doses and post-irradiation time (from 10.8- 47.2%). Interestingly, when only motile spermatozoa from irradiated samples were analysed, only 0.5% were TUNEL positive. CONCLUSION: Motility may be a relevant physiological marker for DNA-intact sperm after exposure of spermatozoa to H(2)O(2) and alpha irradiation. PMID- 11473969 TI - The effect of insemination/injection time on the results of IVF and ICSI. AB - BACKGROUND: The aim of this study was to investigate whether a pre-incubation time between oocyte retrieval and insemination or injection had any effect on the success rate of IVF or intracytoplasmic sperm injection (ICSI). Based on previously published data, many laboratories retain a time interval of several hours between oocyte retrieval and insemination/injection. In our setting, insemination and injection times are dependent only on the laboratory workload. METHODS: Totals of 881 IVF and 432 ICSI cycles performed between 1997 and 1999 were analysed retrospectively. Oocyte retrieval occurred 36 h after human chorionic gonadotrophin administration, and insemination or injection took place 1--7 or 0.5--8 h after oocyte retrieval respectively. RESULTS: No statistically significant differences were found between these time periods and outcome of IVF and ICSI with respect to fertilization rate, embryo quality, implantation rate, abortion and ongoing pregnancy rates, except for the abortion rate after IVF. As this finding may be due to chance and no differences were found in the ongoing pregnancy rates, this finding was considered to be of less importance. CONCLUSIONS: If laboratory control and efficiency demands early insemination or injection, it could be performed without reservation. PMID- 11473970 TI - Morphology of in-vitro matured oocytes: impact on fertility potential and embryo quality. AB - BACKGROUND: The purpose of the present study was to investigate the morphology of in-vitro matured metaphase II (MII) oocytes and to observe if there was a difference in the morphology between polycystic and normal ovaries. Furthermore, the morphology of in-vitro matured MII oocytes was related to their subsequent fertilization and cleavage rates and to embryo quality. METHODS: This retrospective study included 264 MII oocytes obtained in 100 consecutive cycles. Oocyte retrieval was performed transvaginally and cumulus enclosed oocytes were matured for 28--30 h before evaluation. Prior to ICSI, all MII oocytes were graded into three groups according to the number of anomalies: grade I: oocytes without any anomaly (n = 144, 54%), grade II: oocytes with one anomaly (n = 87, 33%) and grade III: oocytes with at least two anomalies (n = 33, 12.5%). RESULTS: Oocyte grades did not differ between women with polycystic ovaries [grades I, II and III respectively: 58/94 (61.7%), 29/94 (30.9%) and 7/94 (7.4%)] and women with normal ovaries [grades I, II and III respectively: 86/170 (50.6%); 58/170 (34%); 26/170 (15.3%)]. Morphology was not related to fertilization rates. The cleavage rate was, however, affected by morphological anomalies (grade I [77/144 (53.5%) versus grade II 33/87 (37.9%) (P = 0.03)], although no significant decrease in cleavage rate could be demonstrated when all grade II and III oocytes were compared with normal oocytes. Significantly more embryos of good quality developed after grade I oocytes [54/144 (37.5%)] compared with those from grade II and grade III oocytes (22/120; P = 0.001). The presence of cytoplasmic abnormalities significantly decreased the cleavage rate (P = 0.04) and also the number of good quality embryos (P < 0.001). CONCLUSION: The in-vitro maturation of oocytes without anomalies yields higher quality embryos, with higher cleavage rates, than those with anomalies. PMID- 11473971 TI - Randomized placebo controlled trial of lofexidine hydrochloride for chronic pelvic pain in women. AB - BACKGROUND: We hypothesised that the orally-active alpha(2)-adrenoceptor agonist lofexidine hydrochloride would ameliorate chronic pelvic pain in women. METHODS: A randomized placebo-controlled parallel group trial was undertaken in the University Hospital Gynaecology Clinic. Women with pelvic pain of at least 6 months duration were eligible, and were randomized using a sealed envelope system to receive up to 600 mg lofexidine hydrochloride twice daily over 8 weeks or placebo. Outcome measures were summary and daily diary visual analog scales for pain (VAS) and a 5 point self rating scale. RESULTS: 9/19 women randomized to lofexidine completed the study compared to 14/20 of those randomized to placebo. Intention-to-treat analysis showed that 4/19 in the lofexidine group achieved 50% or greater reduction in VAS compared with 8/20 in the placebo group (OR 2.5, 95% CI 0.6--10.3). Summary and diary VAS were closely correlated. CONCLUSIONS: Within the limits of a small study with power to detect only a substantial effect, we conclude that lofexidine hydrochloride is not effective for the treatment of chronic pelvic pain. PMID- 11473972 TI - Utero-vaginal anastomosis in women with uterine cervix atresia: long-term follow up and reproductive performance. A study of 18 cases. AB - BACKGROUND: Atresia of the uterine cervix is an uncommon Mullerian anomaly. Total hysterectomy remains the classical treatment of this malformation. The purpose of this study was to evaluate functional results and reproductive performance of women who had conservative surgical procedure. METHODS AND RESULTS: The medical records of 18 patients admitted to our centre between 1969 and 1998 for the treatment of uterine cervix atresia, were reviewed. Seven women had an associated high vaginal aplasia. Fifteen women had a history of abdominal or pelvic surgery before referral, with an unsuccessful attempt at canalization in five cases. Associated pelvic endometriosis or adhesions were observed in 12 cases. The utero vaginal anastomosis procedure was performed successfully in all cases. A secondary stenosis of the anastomosis occurred in one case and this required canalization. Median follow-up after surgery was 4.5 years. Sexual intercourse was satisfactory for the 12 patients who began sexual activity. Ten patients had a pregnancy, four of which resulted in a total of six successful spontaneous pregnancies. Of the six remaining women, five had an evident cause of infertility. CONCLUSIONS: Utero-vaginal anastomosis should be proposed in women with congenital atresia of the uterine cervix, even when it is associated with vaginal aplasia. Early diagnosis and surgery appear necessary to avoid the development of pelvic associated lesions. PMID- 11473973 TI - Laparoscopic myomectomy: predicting the risk of conversion to an open procedure. AB - BACKGROUND: Laparoscopic myomectomy (LM) has some advantages over laparotomy; however, it is reputed to be technically difficult, and the risk of conversion to laparotomy might be an obstacle in using this procedure. The aim of this study was to identify the pre-operative factors affecting the risk of conversion to an open procedure (either laparoscopic assisted myomectomy or laparotomy), and to develop a simple prediction model based on available pre-operative data with the use of multiple logistic regression. METHODS: A total of 426 women presenting with a subserous or intramural myoma measuring 20 mm or more underwent LM between March 1989 and October 1999. Of these patients, 378 had successful LM. Forty eight patients [11.3%, 95% confidence interval (CI) 8.3--14.3] had a conversion to an open procedure. A total of 265 women had adequate pre-operative ultrasonography (US) and were used for the analysis. RESULTS: The best prediction model included four pre-operative factors that were found to be independently related to the risk of conversion: size > or = 50 mm at US (adjusted OR = 10.3; 95% CI = 2.8--37.9), intramural type (adjusted OR = 4.3; 95% CI = 1.3--14.5), anterior location (adjusted OR = 3.4; 95% CI = 1.3-9.0) and pre-operative use of gonadotrophin-releasing hormone (GnRH) agonists (adjusted OR = 5.4; 95% CI = 2.0- 14.2). The regression coefficients were then scaled and rounded to integers to provide an estimate of the risk for conversion. For a given patient with selected characteristics the predicted risk varied from 0--73%. CONCLUSIONS: This prediction model provides a useful tool that enables multiple criteria to be taken into account simultaneously to help select cases for LM. GnRH agonists should been used only in selected cases. US evaluation is essential before performing LM. PMID- 11473974 TI - Prevention of postoperative adhesion formation in rat uterine horn model by nimesulide: a selective COX-2 inhibitor. AB - BACKGROUND: Pelvic surgery is one of the main causes of intraperitoneal (i.p.) adhesions that create various medical problems including pelvic pain, bowel obstructions and female infertility. A rat model was used to investigate the efficacy of nimesulide, a selective cyclooxygenase-2 inhibitor, in the prevention of adhesion formation. METHODS: Fifty Wistar-Albino rats underwent bilateral uterine horn injury with a unipolar cautery. Study groups were as follows: (i) control group, no adjuvant therapy; (ii) i.p. Ringer's lactate group, 2 ml Ringer's lactate solution was instilled i.p.; (iii) i.p. Ringer's lactate plus nimesulide group, 1 ml Ringer's lactate plus 1 ml nimesulide (0.5 mg/ml) were given i.p.; (iv) intramuscular (i.m.) nimesulide group, 1 ml i.m. nimesulide (0.5 mg/ml) was given preoperatively for 5 days; and (v) i.p. nimesulide group, 1 ml nimesulide (0.5 mg/ml) was instilled i.p. At the end of the study all animals were killed, and a standard adhesion scoring system was applied by a blinded examiner. RESULTS: The mean adhesion extent in study groups was as follows: 1.33 +/- 0.76 in control group, 1.40 +/- 0.90 in i.p. Ringer's lactate group, 0.75 +/- 0.70 in i.p. Ringer's lactate plus nimesulide group, 0.25 +/- 0.44 in i.m. nimesulide group and 0.31 +/- 0.70 in i.p. nimesulide group. The mean +/- SD adhesion severities of control, i.p. Ringer's lactate, i.p. Ringer's lactate plus nimesulide, i.m. nimesulide, and i.p. nimesulide groups were 0.58 +/- 0.35, 0.30 +/- 0.41, 0.27 +/- 0.3, 0.12 +/- 0.28 and 0.15 +/- 0.35 respectively. The lowest adhesions were found in the groups treated with nimesulide i.m. and nimesulide i.p. ( P < 0.05). CONCLUSIONS: This study showed that preoperative i.m. or postoperative i.p. administration of nimesulide to the site of injury reduced the formation of postoperative adhesions in a rat uterine horn model. PMID- 11473975 TI - Peritoneal endometriosis: validation of an in-vivo model. AB - BACKGROUND: The current medical treatment of endometriosis, a common gynaecological disease, is still associated with a high recurrence rate. To establish an appropriate in-vivo model to evaluate new therapeutic strategies we validated the nude mouse model for the intraperitoneal cultivation of human endometrial tissue. METHODS: Human endometrium of the proliferative phase was implanted into the peritoneal cavity of normal cycling and ovariectomized athymic mice and of cycling non-obese diabetic (NOD)-severe combined immuno-deficiency (SCID) mice. Morphology, proliferation, differentiation, and angiogenesis in the ectopic endometrium at different time points after implantation was investigated. RESULTS: Adhesion of endometrial fragments was observed from day 2 onwards. The lesions persisted for up to 28 days revealing a well preserved glandular morphology. The glandular epithelium maintained cytokeratin expression even after 14 days of culture. With progressing culture, glands exhibited vimentin staining in combination with a decrease of surrounding stromal cells. Proliferation of glandular epithelium could be demonstrated throughout the investigated period of 28 days, whereas expression of oestrogen and progesterone receptors was maintained only in endometriotic lesions grown in cycling but not in ovariectomized mice. Neoangiogenesis occurred from day 4 onwards, independent from the intraperitoneal localization of the ectopic lesions. CONCLUSIONS: This in-vivo model is a promising tool to test the effect of compounds such as different hormone agonists/antagonists or anti-angiogenic factors to develop new therapeutic concepts in endometriosis. PMID- 11473976 TI - Magnetic resonance imaging of the human cervix: a study of the effects of prostaglandins in the first trimester. AB - BACKGROUND: To establish that magnetic resonance imaging (MRI) can provide an objective measurement of the biophysical state of the cervix and to measure the response of the cervix to prostaglandins in the first trimester of pregnancy. METHODS: A comparative study, with each patient serving as her own control, was carried out on 10 primigravid women with 49--84 days amenorrhoea undergoing surgical termination of pregnancy. Each woman had two MRI scans of the uterus and cervix. The first scan was performed prior to administration of prostaglandins and the second scan 2.5--3 h following an 800 microg dose of misoprostol given vaginally. Cervical change was assessed by measurement of cervical length, transverse diameter of the cervix, diameter of the internal os and cervical stromal signal intensity. RESULTS: A significant difference in cervical length (P = 0.012), transverse diameter (P = 0.001) and diameter of the internal os (P = 0.008) was detected following the administration of misoprostol. In five women a significant change in one or both components of the cervical stromal signal was detectable and in five women no change could be demonstrated. CONCLUSION: MRI is capable of detecting changes in the physical parameters of the cervix following administration of prostaglandin. However, the changes detected in the stromal signal intensity are less consistent. The potential of MRI as a clinical tool to monitor cervical changes in a variety of clinical situations warrants further investigation. PMID- 11473977 TI - Is voluntary abortion a seasonal disorder of mood? AB - BACKGROUND: Depressive mood and suicides are more frequent in women seeking voluntary abortion and occur in a seasonal rhythmic fashion. Whether voluntary abortion shows a similar seasonal rhythm was investigated in this study. METHODS: A 4-year analysis was performed on the database of the National Institute of Statistics (ISTAT) (508,130 abortions) and on the medical records of our institute (3463 abortions). The ratio of voluntary abortions to the number of vital pregnancies (terminated with birth and voluntary abortion) present at the third month of gestation (8--12 weeks) was evaluated. Analyses were carried out by the periodogram method. RESULTS: The rate of voluntary abortions showed a seasonal rhythm with an amplitude of 6.1--6.7% and peaked in May (+/-38 days). The national frequency of female suicides, obtained from the same ISTAT database, showed a similar rhythm, with an amplitude of 11.1% and maximal rate in June (+/ 37 days). CONCLUSIONS: The present data show a seasonal rhythm in the rate of voluntary abortion, which is almost identical to that of female suicides. This link suggests common provocative mechanisms and may indicate common preventative measures. PMID- 11473978 TI - Psychosocial characteristics of infertile couples: a study by the 'Heidelberg Fertility Consultation Service'. AB - BACKGROUND: The aim of the study was to identify differences in psychological characteristics between couples with fertility disorders, especially idiopathic infertility, and a representative sample. MATERIAL AND METHODS: A total of 564 couples was examined using psychological questionnaires pertaining to sociodemographic factors, motives for wanting a child, dimensions of life satisfaction and couple relationships, physical and psychic complaints, and a personality inventory. RESULTS: Specific to our sample was the high educational level of the couples, and the large number with idiopathic infertility (27% of all diagnoses). There were no remarkable differences in psychological variables between the infertile couples and a representative sample, except that the infertile women showed higher scores on the depression and anxiety scales. Couples with idiopathic infertility showed no remarkable differences in the questionnaire variables compared with couples with other medical diagnoses of infertility. CONCLUSIONS: A typical psychological profile for infertile couples could not be identified using standardized psychometric rating methods. This may be an effect of the specific characteristics of our sample. For some couples, the infertility crisis can be seen as a cumulative trauma, which indicates that these couples have a marked need for infertility counselling. PMID- 11473979 TI - The effect of an infertility diagnosis on the distress, marital and sexual satisfaction between husbands and wives in Taiwan. AB - BACKGROUND: Few studies have explored the effect of a gender-specific infertility diagnosis on the responses of couples in Taiwan. The purpose of this research was to compare the differences in distress, marital and sexual satisfaction in husbands and wives based on an infertility diagnosis. METHODS: Three structured questionnaires were used. RESULTS: Female members of couples in which both partners were infertile expressed less marital and sexual satisfaction than their husbands. No differences in marital and sexual satisfaction were found between wives and husbands with unexplained infertility. Only wives with a diagnosed female infertility expressed higher distress to infertility than their husbands. Although no differences in psychosocial responses were found among husbands, regardless of the diagnosis, wives with a diagnosed female infertility experienced higher distress in self-esteem and less satisfaction in acceptance by in-laws than wives experiencing a diagnosed male infertility. CONCLUSIONS: These findings suggest that the diagnosis of infertility is an important factor in assessing the differences in infertility distress and marital and sexual satisfaction between husbands and wives. Health professionals can explain the gender differences when counselling infertile couples and encourage them to share each other's feelings, which may help couples to cope with the communication problems they may experience. PMID- 11473980 TI - Contribution of environmental factors to the risk of male infertility. AB - BACKGROUND: An increasing number of reports suggest that chemical and physical agents in the environment, introduced and spread by human activity, may affect male fertility in humans. We investigated the relationships between exposure to environmental agents and seminal characteristics, and the concentrations of reproductive hormones in the serum of men seeking infertility treatment. METHODS: We studied 225 male partners from consecutively recruited couples, who had their first infertility consultation between 1995 and 1998, in the Litoral Sur region of Argentina, one of the most productive farming regions in the world. RESULTS: A multivariate logistic regression model showed that exposure to pesticides and solvents is significantly associated with sperm threshold values well below the limit for male fertility. We also found that men exposed to pesticides had higher serum oestradiol concentrations, and that men exposed to solvents had lower LH concentrations than non-exposed men. All of these effects were greater in men with primary infertility than in men with secondary infertility. CONCLUSION: We have shown that environmental factors contribute to the severity of infertility, and that this may worsen the effects of pre-existing genetic or medical risk factors. PMID- 11473981 TI - Blood pressure during pregnancy, sex ratio of offspring and reproductive hormone concentrations. PMID- 11473982 TI - Vitrification of embryos and oocytes with 5.5 mol/l ethylene glycol and 1.0 mol/l sucrose. PMID- 11473985 TI - Certain forms of morphological anomalies of spermatozoa may reflect chromosomal aneuploidies. PMID- 11473986 TI - Spontaneous pregnancies in severe oligoasthenozoospermia. PMID- 11473987 TI - The COPE Report 1999. Guidelines on good publication practice. PMID- 11473988 TI - Detection of precytopathic effect of enteroviruses in clinical specimens by centrifugation-enhanced antigen detection. AB - Rapid enterovirus detection is important for decisions about antibiotic administration and length of hospital stay. The efficacy of rapid antigen detection-cell culture amplification (Ag-CCA) was evaluated with monoclonal antibodies (MAbs) 5-D8/1 (DAKO) and Pan-Enterovirus clone 2E11 (Chemicon) with 10 poliovirus, echovirus, and coxsackievirus type A and B stock isolates and College of American Pathologists check samples. By using Ag-CCA technology, MAb 2E11 was more sensitive than 5-D8/1 at detecting a greater number of stock isolates at or past tube (cytopathic effect [CPE]) culture (TC) end points. The efficacy of Ag CCA in the clinical setting was subsequently confirmed with 273 consecutively freshly collected nasopharyngeal aspirate or swab specimens, rectal swab, and cerebrospinal fluid specimens during the 1999 enterovirus season. All specimens were tested by Ag-CCA in parallel with rhesus monkey kidney (RhMk), MRC-5, and A549 conventional TCs. Approximately 60% of field specimens were additionally tested with Hep-2 and HNK conventional TCs. Sixty-two percent of the clinical specimens tested were Ag-CCA positive after 48 h. Among 51 isolates, the mean time to CPE or culture confirmation was 5.5 days (range, 2 to 18 days). After 48 h, Ag-CCA achieved sensitivity, specificity, and positive and negative predictive values of 62, 100, 100, and 93%, respectively. During the same period, TC-CPE displayed test parameters of 12, 100, 100, and 85%, respectively. After 5 days, the sensitivity and specificity of Ag-CCA increased to 92 and 98%, respectively. Within the same period, isolation attained sensitivity and specificity of 52 and 100%, respectively. Although Ag-CCA displayed slightly reduced sensitivity and reduced specificity compared with conventional cell culture after 14 days, the markedly superior 48-h enterovirus Ag-CCA detection rate supports incorporation of this assay into the routine clinical setting. PMID- 11473989 TI - Genetic variation among hospital isolates of methicillin-sensitive Staphylococcus aureus: evidence for horizontal transfer of virulence genes. AB - Staphylococcus aureus strains often carry in their genomes virulence genes that are not found in all strains and that may be carried on discrete genetic elements. Strains also differ in that they carry one of four classes of an accessory gene regulator (agr) locus, an operon that regulates virulence factor expression and that has been proposed to be a therapeutic target. To look at their distribution among hospital strains, we investigated 38 methicillin sensitive S. aureus isolates, classifying the isolates by agr class and screening them for the presence and restriction fragment length polymorphisms (RFLPs) of 12 core and 14 accessory virulence genes. Twenty-three (61%) were agr class I, 10 (26%) were agr class II, and 5 (13%) were agr class III. None were agr class IV. The S. aureus strains had distinguishable RFLP profiles, although clusters of isolates with clearly related core gene profiles were found among our strains, including all five agr class III strains, two sets of six strains within agr class I, and six strains within agr class II. Within these clusters there was evidence of horizontal acquisition and/or loss of multiple accessory virulence genes. Furthermore, two isolates from the same patient were identical except for the presence of the sea gene, indicating that movement of mobile elements may occur in vivo. Several strong correlations with the carriage of virulence genes between strains were seen, including a positive correlation between tst and agr class III and negative correlations between tst and lukE-splB and between lukE splB and seg-sei. This suggests that the core genome or the presence of accessory genetic elements within a strain may influence acquisition and loss of other elements encoding virulence genes. PMID- 11473990 TI - 16S/23S rRNA intergenic spacer regions for phylogenetic analysis, identification, and subtyping of Bartonella species. AB - Species of the genus Bartonella are currently recognized in growing numbers and are involved in an increasing variety of human diseases, mainly trench fever, Carrion's disease, bacillary angiomatosis, endocarditis, cat scratch disease, neuroretinitis, and asymptomatic bacteremia. Such a wide spectrum of infections makes it necessary to develop species and strain identification tools in order to perform phylogenetic and epidemiological studies. The 16S/23S rRNA intergenic spacer region (ITS) was sequenced for four previously untested species, B. vinsonii subsp. arupensis, B. tribocorum, B. alsatica, and B. koehlerae, as well as for 28 human isolates of B. quintana (most of them from French homeless people), six human or cat isolates of B. henselae, five cat isolates of B. clarridgeiae, and four human isolates of B. bacilliformis. Phylogenetic trees inferred from full ITS sequences of the 14 recognized Bartonella species using parsimony and distance methods revealed high statistical support, as bootstrap values were higher than those observed with other tested genes. Five well supported lineages were identified within the genus and the proposed phylogenetic organization was consistent with that resulting from protein-encoding gene sequence comparisons. The ITS-derived phylogeny appears, therefore, to be a useful tool for investigating the evolutionary relationships of Bartonella species and to identify Bartonella species. Further, partial ITS amplification and sequencing offers a sensitive means of intraspecies differentiation of B. henselae, B. clarridgeiae, and B. bacilliformis isolates, as each strain had a specific sequence. The usefulness of this approach in epidemiological investigations should be highlighted. Among B. quintana strains, however, the genetic heterogeneity was low, as only three ITS genotypes were identified. It was nevertheless sufficient to show that the B. quintana population infecting homeless people in France was not clonal. PMID- 11473991 TI - Comparison of a multiplex reverse transcription-PCR-enzyme hybridization assay with conventional viral culture and immunofluorescence techniques for the detection of seven viral respiratory pathogens. AB - A multiplex reverse transcription-PCR-enzyme hybridization assay (RT-PCR-EHA; Hexaplex; Prodesse Inc., Waukesha, Wis.) was used for the simultaneous detection of human parainfluenza virus types 1, 2, and 3, influenza virus types A and B, and respiratory syncytial virus types A and B. One hundred forty-three respiratory specimens from 126 patients were analyzed by RT-PCR-EHA, and the results were compared to those obtained by conventional viral culture and immunofluorescence (IF) methods. RT-PCR-EHA proved to be positive for 17 of 143 (11.9%) specimens, whereas 8 of 143 (5.6%) samples were positive by viral culture and/or IF. Eight samples were positive by both RT-PCR-EHA and conventional methods, while nine samples were RT-PCR-EHA positive and viral culture and IF negative. Eight of the nine samples with discordant results were then independently tested by a different multiplex RT-PCR assay for influenza virus types A and B, and all eight proved to be positive. In comparison to viral culture and IF methods, RT-PCR-EHA gave a sensitivity and a specificity of 100 and 93%, respectively. Since RT-PCR-EHA was able to detect more positive samples, which would otherwise have been missed by routine methods, we suggest that this multiplex RT-PCR-EHA provides a highly sensitive and specific means of diagnostic detection of major respiratory viruses. PMID- 11473992 TI - Characterization of mutations in the rpoB gene associated with rifampin resistance in Rhodococcus equi isolated from foals. AB - Treatment with a combination of erythromycin and rifampin has considerably improved survival rates of foals and immunocompromised patients suffering from severe pneumonia caused by Rhodococcus equi. Frequently, because of monotherapy, emergence of rifampin-resistant strains has been responsible for treatment failure. Using consensus oligonucleotides, we have amplified and sequenced the rifampin resistance (Rif(r))-determining regions of 12 rifampin-resistant R. equi strains isolated from three foals and of mutants selected in vitro from R. equi ATCC 3701, a rifampin-susceptible strain. The deduced amino acid sequences compared to those of four rifampin-susceptible R. equi strains showed several types of mutations. In 3 of the 10 strains isolated from one foal, His526Asn (Escherichia coli numbering) and Asp516Val mutations were associated with low level resistance (rifampin MIC, 2 to 8 microg/ml), whereas His526Asp conferred high-level resistance (rifampin MIC, 128 microg/ml) in the 7 remaining strains. In strains from the two other foals, His526Asp and Ser531Leu mutations were found to be associated with high-level and low-level resistance, respectively. The in vitro mutants, highly resistant to rifampin, harbored His526Tyr and His526Arg substitutions. As described in other bacterial genera, His526, Ser531, and Asp516 are critical residues for rifampin resistance in R. equi, and the resistance levels are dependent on both the location and the nature of the substitution. PMID- 11473993 TI - Molecular and antigenic comparison of Ehrlichia canis isolates from dogs, ticks, and a human in Venezuela. AB - We previously culture isolated a strain of Ehrlichia canis, the causative agent of canine ehrlichiosis, from a human in Venezuela. In the present study, we examined whether dogs and ticks are infected with E. canis in Venezuela and, if so, whether this is the same strain as the human isolate. PCR analysis using E. canis-specific primers revealed that 17 of the 55 dog blood samples (31%) and all three pools of four Rhipicephalus sanguineus ticks each were positive. An ehrlichial agent (Venezuelan dog Ehrlichia [VDE]) was isolated and propagated in cell culture from one dog sample and was further analyzed to determine its molecular and antigenic characteristics. The 16S rRNA 1,408-bp sequence of the new VDE isolate was identical to that of the previously reported Venezuelan human Ehrlichia isolate (VHE) and was closely related (99.9%) to that of E. canis Oklahoma. The 5' (333-bp) and 3' (653-bp) sequences of the variable regions of the 16S rRNA genes from six additional E. canis-positive dog blood specimens and from three pooled-tick specimens were also identical to those of VHE. Western blot analysis of serum samples from three dogs infected with VDE by using several ehrlichial antigens revealed that the antigenic profile of the VDE was similar to the profiles of VHE and E. canis Oklahoma. Identical 16S rRNA gene sequences among ehrlichial organisms from dogs, ticks, and a human in the same geographic region in Venezuela and similar antigenic profiles between the dog and human isolates suggest that dogs serve as a reservoir of human E. canis infection and that R. sanguineus, which occasionally bites humans residing or traveling in this region, serves as a vector. This is the first report of culture isolation and antigenic characterization of an ehrlichial agent from a dog in South America, as well as the first molecular characterization of E. canis directly from naturally infected ticks. PMID- 11473994 TI - Detection of dengue viral RNA using a nucleic acid sequence-based amplification assay. AB - Faster techniques are needed for the early diagnosis of dengue fever and dengue hemorrhagic fever during the acute viremic phase of infection. An isothermal nucleic acid sequence-based amplification (NASBA) assay was optimized to amplify viral RNA of all four dengue virus serotypes by a set of universal primers and to type the amplified products by serotype-specific capture probes. The NASBA assay involved the use of silica to extract viral nucleic acid, which was amplified without thermocycling. The amplified product was detected by a probe hybridization method that utilized electrochemiluminescence. Using normal human plasma spiked with dengue viruses, the NASBA assay had a detection threshold of 1 to 10 PFU/ml. The sensitivity and specificity of the assay were determined by testing 67 dengue virus-positive and 21 dengue virus-negative human serum or plasma samples. The "gold standard" used for comparison and evaluation was the mosquito C6/36 cell culture assay followed by an immunofluorescent assay. Viral infectivity titers in test samples were also determined by a direct plaque assay in Vero cells. The NASBA assay was able to detect dengue viral RNA in the clinical samples at plaque titers below 25 PFU/ml (the detection limit of the plaque assay). Of the 67 samples found positive by the C6/36 assay, 66 were found positive by the NASBA assay, for a sensitivity of 98.5%. The NASBA assay had a specificity of 100% based on the negative test results for the 21 normal human serum or plasma samples. These results indicate that the NASBA assay is a promising assay for the early diagnosis of dengue infections. PMID- 11473995 TI - Identification of 54 mycobacterial species by PCR-restriction fragment length polymorphism analysis of the hsp65 gene. AB - A total of 121 reference and clinical strains of both slowly and rapidly growing mycobacteria belonging to 54 species were studied for restriction fragment length polymorphism of a PCR-amplified 439-bp segment of the gene encoding the 65-kDa heat shock protein. Restriction digests were separated by 10% polyacrylamide gel electrophoresis (PAGE). By including a size standard in each sample, the restriction fragment profile was calculated using a computer-aided comparison program. An algorithm describing these 54 species (including 22 species not previously described) is proposed. We found that this assay based on 10% PAGE provided a more precise estimate than that based on agarose gel electrophoresis of the real size of restriction fragments as deduced from the sequence analysis and allowed identification of mycobacteria whose PCR-restriction fragment length polymorphism analysis patterns were unequivocally identified by fragments shorter than 60 bp. PMID- 11473996 TI - Antibody-secreting cell responses to rotavirus proteins in gnotobiotic pigs inoculated with attenuated or virulent human rotavirus. AB - Because of their similarities to infants in mucosal immune responses and their susceptibility to human rotavirus (HRV) diarrhea, gnotobiotic pigs provide a useful model for rotaviral disease. In this study, we performed quantitative enzyme-linked immunospot (ELISPOT) assays to measure local and systemic isotype specific antibody-secreting cell (ASC) responses to individual structural (VP4, VP6, and VP7) and nonstructural (NSP3 and NSP4) proteins of Wa HRV. The Spodoptera frugiperda cells expressing each recombinant baculovirus HRV protein were formalin fixed and used as antigen for ELISPOT assays. Neonatal gnotobiotic pigs were orally inoculated once with virulent Wa (WaV) or three times with attenuated Wa (WaA) HRV or mock inoculated (Mock) and then were challenged with virulent Wa (WaV/PC) 28 days after the first inoculation. The ASCs from intestinal and systemic lymphoid tissues of pigs from each group were quantitated by ELISPOT assay at the day of challenge, at postinoculation day 28 (WaV, WaA, and Mock) or at postchallenge day (PCD) 7 (WaV+WaV/PC, WaA+WaV/PC, and Mock+WaV/PC). In all virus-inoculated pigs, regardless of the inoculum, lymphoid tissue, or isotype, VP6 induced the highest numbers of ASCs, followed by VP4; ASCs specific for VP7, NSP3, and NSP4 were less numerous. At challenge, total HRV and HRV protein-specific immunoglobulin A (IgA) and IgG ASCs in intestinal lymphoid tissues were significantly greater in WaV- than in WaA-inoculated pigs, and WaV pigs were fully protected against diarrhea postchallenge, whereas the WaA pigs were partially protected. At PCD 7, there were no significant differences in ASC numbers for any HRV proteins between the WaV+WaV/PC and WaA+WaV/PC groups. PMID- 11473997 TI - Characterization and analysis of a stable serotype-associated membrane protein (P30) of Mycoplasma agalactiae. AB - The gene for a 30-kDa immunodominant antigen, P30, of Mycoplasma agalactiae was cloned from type strain PG2 and expressed in Escherichia coli. P30 is encoded on a monocistronic operon determined by two -10 boxes and a possible -35 region constituting the potential promoter, and a transcription termination site. The gene for the 266-amino-acid protein is preceded by a polypurine-rich region designed as the consensus sequence for a ribosome-binding site. Analysis of the amino acid sequence of P30 revealed the presence of a recognition site for a prokaryotic signal peptidase II at amino acid (aa) 24, indicating that P30 is a transmembrane protein. Moreover, Triton X-114 phase partitioning of M. agalactiae PG2 total antigen revealed that P30 is strongly hydrophobic and hence a possible membrane component. Immunoblot analysis using the monospecific polyclonal anti P30-His serum indicated that P30 is specific to M. agalactiae. Furthermore, PCR amplification with specific primers for p30 and Southern blot analysis revealed the presence of the gene in all M. agalactiae strains tested and its absence in the other mycoplasma species. Among 27 strains of M. agalactiae studied, 20 strains belonging to the common serotypes A to D, including PG2, expressed P30 or part of it as detected by the monospecific polyclonal anti-P30 antibodies. The other seven strains belonging to the rarely isolated serotypes E to H were negative for P30. The p30 gene was sequenced in 15 strains of M. agalactiae, 10 of which expressed P30 or at least part of it and 5 of which did not express P30. The negative strains carried mutations in both -10 boxes of the promoters. These mutations seem to be responsible for the lack of P30 expression in these strains. Analysis of sera from sheep that were experimentally infected with M. agalactiae revealed that P30 induced a strong and persistent immune response which was still very high two months after infection. In contrast, currently used enzyme-linked immunosorbent assay serology gave only low titers. PMID- 11473998 TI - Improved antimicrobial interventions have benefits. AB - Studies have shown benefits to patients from improved interventions involving antimicrobial therapy. The purpose of the present study was to evaluate prospectively the impact of improved interventions by (i) the use of TheraTrac 2, a computer software program which electronically links susceptibility testing results immediately to the pharmacy and alerts pharmacists of potential interventions, and (ii) the education of pharmacists involving microbiologic topics. The study group had the new intervention program. The control group had interventions performed the way that they had previously been done by manually reviewing hard copies of susceptibility testing data. In a 5-month period, all inpatients whose last names began with A to K were the study group; inpatients whose last names began with L to Z were controls. Three analyses were done; one analysis (analysis A) involved only patients with interventions, one analysis (analysis B) involved all patients for whom antimicrobial testing was done and who were matched for diagnosis-related groups (DRGs), regardless of whether an intervention occurred, and one analysis (analysis C) involved these DRG-matched patients by using severity-adjusted data. In analysis A, the study group had a 4.8% decreased rate of mortality, an average of a 16.5-day decreased length of stay per patient, and $20,886 decreased variable direct costs per patient. None of these differences was statistically significant. In analysis B, the study patients had a 1.2% higher mortality rate (P = 0.741), an average of a 2.7-day decreased length of stay per patient (P = 0.035), and $2,626 decreased variable direct costs per patient (P = 0.008). In analysis C, the study patients had a 1.4% lower mortality rate, a 1.2-day decreased length of stay per patient, and $1,466 decreased variable direct costs per patient. In conclusion, the institution of this program caused substantial cost savings. PMID- 11473999 TI - Clinical isolates of non-O157 Shiga toxin-producing Escherichia coli: serotypes, virulence characteristics, and molecular profiles of strains of the same serotype. AB - All human Shiga toxin-producing Escherichia coli (STEC) non-O157 strains (n = 56) isolated in Finland from 1990 to August 2000 were characterized for the O:H serotype, stx(1) and stx(2) genes, production of enterohemolysin, and sensitivity to 12 antimicrobial agents. Strains of the same serotype were genotyped by pulsed field gel electrophoresis (PFGE) after XbaI restriction of total DNA. The 56 non O157 isolates belonged to 29 serotypes. Two of the serotypes (O102:H7 and OX181:H49) have not previously been described as being associated with STEC infections in humans or isolated from animals. Thirty-four strains (61%) within seven serotypes (O103:H2 [14 isolates], O26:H11 [6 isolates], O145:H28 [4 isolates], O145:HNM [3 isolates], O15:HNM [3 isolates], OX174:H21 [2 isolates], and O Rough:HNM [2 isolates]) were represented by more than one isolate. Of these strains, O103:H2 isolates were divided into seven, O26:H11 isolates were divided into four, and the rest within a serotype were divided into two genotypes in PFGE. In PCR, 31 (55%) of the 56 strains were positive for the stx(2) gene only and 24 strains (43%) were positive for stx(1) only. One strain (O43:H2) carried both stx(1) and stx(2). Forty-two strains (75%) produced enterohemolysin, and 39 strains (70%) possessed the eae gene. Of the latter 39 strains, 36 (92%) were enterohemolytic, whereas only 6 (35%) of the 17 isolates lacking the eae gene were enterohemolytic (P < 0.001). The majority of the strains (44 strains, 79%) were sensitive to all 12 antimicrobials tested. Of the 56 strains, 20 (36%) were associated with small family outbreaks in nine families and 14 (25%) were associated with recent travel abroad. PMID- 11474000 TI - Detection and quantification of infectious hypodermal and hematopoietic necrosis virus and white spot virus in shrimp using real-time quantitative PCR and SYBR Green chemistry. AB - A rapid and highly sensitive real-time PCR detection and quantification method for infectious hypodermal and hematopoietic necrosis virus (IHHNV), a single stranded DNA virus, and white spot virus (WSV), a double-stranded DNA (dsDNA) virus infecting penaeid shrimp (Penaeus sp.), was developed using the GeneAmp 5700 sequence detection system coupled with SYBR Green chemistry. The PCR mixture contains a fluorescence dye, SYBR Green, which upon binding to dsDNA exhibits fluorescence enhancement. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. A linear relationship was observed between the amount of input plasmid DNA and cycle threshold (C(T)) values over a range of 1 to 10(5) copies of the viral genome. To control the variation in sampling and processing among samples, the shrimp beta-actin gene was amplified in parallel with the viral DNA. The C(T) values of IHHNV- and WSV-infected samples were used to determine absolute viral copy numbers from the standard C(T) curves of these viruses. For each virus and its beta-actin control, the specificity of amplification was monitored by using the dissociation curve of the amplified product. Using genomic DNA as a template, SYBR Green PCR was found to be 100- to 2000-fold more sensitive than conventional PCR, depending on the virus, for the samples tested. The results demonstrate that SYBR Green PCR can be used as a rapid and highly sensitive detection and quantification method for shrimp viruses and that it is amenable to high-throughout assay. PMID- 11474001 TI - Evaluation of methods for detection of toxins in specimens of feces submitted for diagnosis of Clostridium difficile-associated diarrhea. AB - Clostridium difficile is the principal pathogen associated with hospital-acquired acute diarrheal disease. We have evaluated the performances of six approaches for diagnosis of C. difficile-associated diarrhea (CDAD). Consecutive stool specimens (n = 200) from 133 patients were examined by cytotoxin assay, by culture of C. difficile on cycloserine-cefoxitin-fructose agar, and by toxin detection using four rapid immunoassay systems (Oxoid Toxin A test, ImmunoCard Toxin A test, TechLab Tox A/B II test, and Premier Toxins A&B test). A diagnosis of CDAD was established for 35 (27%) patients (representing 29% of specimens). The adjusted sensitivity and specificity of the methods were, respectively, 98 and 99% for the cytotoxin assay, 54 and 99% for ImmunoCard, 50 and 98% for Oxoid, 79 and 98% for TechLab, 80 and 98% for Premier, and 57 and 100% for culture. The TechLab and Premier assays are acceptable tests for diagnosis of CDAD but are not equivalent to the cytotoxin assay. PMID- 11474002 TI - Assessment, by transcription-mediated amplification, of virologic response in patients with chronic hepatitis C virus treated with peginterferon alpha-2a. AB - Transcription-mediated amplification (TMA) is an isothermal, autocatalytic target amplification method which has the potential to detect less than 50 hepatitis C virus (HCV) RNA copies/ml (10 IU/ml). The TMA assay was used to assess the presence of residual HCV RNA in plasma from patients treated with polyethylene glycol-modified interferon alpha-2a (peginterferon alpha-2a) who showed a virologic relapse after the end of therapy. Stored end-of-treatment and end-of follow-up plasma samples from 177 of 267 patients treated with peginterferon alpha-2a (S. Zeuzem et al., N. Engl. J. Med. 343:1666--1672, 2000) were available for retesting by TMA. Plasma samples from patients in the same study who exhibited virologic relapse after treatment with standard interferon alpha-2a served as controls. Virologic response during the trial was defined as HCV RNA that was undetectable using a PCR-based test system with a sensitivity of 50 IU/mL (Cobas Amplicor HCV version 2.0) and was compared with TMA-based retesting results (VERSANT HCV RNA Qualitative Assay). Residual HCV RNA was detected in 4 of 60 cases (7%) by the TMA technology in end-of-treatment plasma samples from patients who relapsed after receiving peginterferon alpha-2a and in 6 of 18 patients (33%) following therapy with standard interferon alpha-2a. For peginterferon alpha-2a-treated patients with sustained virologic response, HCV RNA was detectable by TMA in end-of-treatment samples in 3 of 78 cases but in none of the end-of-follow-up samples. For all end-of-treatment and end-of-follow up plasma samples of virologic nonresponders, a complete concordance between the PCR-based assay and TMA was observed. In conclusion, in patients with virologic relapse after the end of therapy, according to PCR, who were treated with peginterferon alpha-2a or standard interferon alpha-2a, residual HCV RNA was detectable in end-of-treatment samples by the TMA-based assay in 7 or 33% of cases, respectively. The lower rate of residual HCV RNA detection by TMA for patients treated with peginterferon alpha-2a than that for patients treated with standard interferon alpha-2a may be due to the maintained antiviral pressure of the long-acting peginterferon alpha-2a at the end-of-treatment visit. PMID- 11474003 TI - Quantitation of varicella-zoster virus DNA in patients with Ramsay Hunt syndrome and zoster sine herpete. AB - Varicella-zoster virus (VZV) reactivation causes facial nerve palsy in Ramsay Hunt syndrome (RHS) and zoster sine herpete (ZSH) with and without zoster rash, respectively. In the present study, we analyzed the VZV DNA copy number in saliva samples from 25 patients with RHS and 31 patients with ZSH using a TaqMan PCR assay to determine differences in the viral load between the two diseases. VZV copy number in saliva peaked near the day of the appearance of zoster in patients with RHS. Consequently, VZV DNA was less frequently detected in patients with RHS who exhibited facial palsy several days after the appearance of zoster. These findings suggest that the VZV load in saliva samples reflects the kinetics of viral reactivation in patients with RHS. In addition, VZV DNA was equally detected in saliva from patients with RHS and ZSH, and there was no significant difference in the highest viral copy number between patients with RHS and those with ZSH. The VZV load does not appear to reflect a major difference between RHS and ZSH. PMID- 11474004 TI - Transmission of hepatitis C virus in a gynecological surgery setting. AB - A cluster of hepatitis C virus (HCV) infections among gynecological patients who underwent surgical intervention in the same setting is described. An epidemiological investigation was conducted to identify the cases, the likely source of infection, and the route of transmission. Four recent HCV infections were identified. Based on molecular fingerprinting analysis and epidemiological investigation, transmission between the putative source patient (an HCV-positive woman who was the first patient of the surgical session) and outbreak patients was highly suggestive. All patients, including the source patient, were infected with HCV type 1b. Molecular characterization of HCV clones by sequence analysis of both structural envelope regions (20 clones from the source patient and 58 from the outbreak patients) and the nonstructural NS5 region of the viral genome (12 clones from the source patient and 32 from the outbreak patients) showed close homology between the viral isolates from the source and those from the outbreak patients that was higher than that observed between the viral isolates from the source and those from four unrelated, HCV type 1b-infected patients from the same geographical area (in the latter case, 33 clones were sequenced for the envelope regions and 30 were sequenced for the NS5 region). The mean percent divergence between clones was 4.69 for the envelope and 3.71 for the NS5 region in the source patient and the outbreak patients compared with 6.76 (P = 0.001) and 5.22 (P = 0.01) in the source patient and control patients, respectively. Among the risk factors investigated, only that of having undergone surgery in the morning session of the same day reached statistical significance (P = 0.003). The investigation showed that the source patient and outbreak patients shared only the administration of propofol in multidose vials. The study documents the risk of nosocomial transmission of HCV and the importance of infection control procedures in the operating room and highlights the crucial role of molecular strategies, especially sequence-based phylogenetic analysis of cloned viral isolates, in the investigation of HCV outbreaks. PMID- 11474005 TI - Characterization of clinical isolates of Klebsiella pneumoniae from 19 laboratories using the National Committee for Clinical Laboratory Standards extended-spectrum beta-lactamase detection methods. AB - Extended-spectrum beta-lactamases (ESBLs) are enzymes found in gram-negative bacilli that mediate resistance to extended-spectrum cephalosporins and aztreonam. In 1999, the National Committee for Clinical Laboratory Standards (NCCLS) published methods for screening and confirming the presence of ESBLs in Klebsiella pneumoniae, Klebsiella oxytoca, and Escherichia coli. To evaluate the confirmation protocol, we tested 139 isolates of K. pneumoniae that were sent to Project ICARE (Intensive Care Antimicrobial Resistance Epidemiology) from 19 hospitals in 11 U.S. states. Each isolate met the NCCLS screening criteria for potential ESBL producers (ceftazidime [CAZ] or cefotaxime [CTX] MICs were > or =2 microg/ml for all isolates). Initially, 117 (84%) isolates demonstrated a clavulanic acid (CA) effect by disk diffusion (i.e., an increase in CAZ or CTX zone diameters of > or =5 mm in the presence of CA), and 114 (82%) demonstrated a CA effect by broth microdilution (reduction of CAZ or CTX MICs by > or =3 dilutions). For five isolates, a CA effect could not be determined initially by broth microdilution because of off-scale CAZ results. However, a CA effect was observed in two of these isolates by testing cefepime and cefepime plus CA. The cefoxitin MICs for 23 isolates that failed to show a CA effect by broth microdilution were > or =32 microg/ml, suggesting either the presence of an AmpC type beta-lactamase or porin changes that could mask a CA effect. By isoelectric focusing (IEF), 7 of the 23 isolates contained a beta-lactamase with a pI of > or =8.3 suggestive of an AmpC-type beta-lactamase; 6 of the 7 isolates were shown by PCR to contain both ampC-type and bla(OXA) genes. The IEF profiles of the remaining 16 isolates showed a variety of beta-lactamase bands, all of which had pIs of < or =7.5. All 16 isolates were negative by PCR with multiple primer sets for ampC-type, bla(OXA), and bla(CTX-M) genes. In summary, 83.5% of the K. pneumoniae isolates that were identified initially as presumptive ESBL producers were positive for a CA effect, while 5.0% contained beta-lactamases that likely masked the CA effect. The remaining 11.5% of the isolates studied contained beta lactamases that did not demonstrate a CA effect. An algorithm based on phenotypic analyses is suggested for evaluation of such isolates. PMID- 11474006 TI - Detection and identification of fungal pathogens by PCR and by ITS2 and 5.8S ribosomal DNA typing in ocular infections. AB - The goal of this study was to determine whether sequence analysis of internal transcribed spacer/5.8S ribosomal DNA (rDNA) can be used to detect fungal pathogens in patients with ocular infections (endophthalmitis and keratitis). Internal transcribed spacer 1 (ITS1) and ITS2 and 5.8S rDNA were amplified by PCR and seminested PCR to detect fungal DNA. Fifty strains of 12 fungal species (yeasts and molds) were used to test the selected primers and conditions of the PCR. PCR and seminested PCR of this region were carried out to evaluate the sensitivity and specificity of the method. It proved possible to amplify the ITS2/5.8S region of all the fungal strains by this PCR method. All negative controls (human and bacterial DNA) were PCR negative. The sensitivity of the seminested PCR amplification reaction by DNA dilutions was 1 organism per PCR, and the sensitivity by cell dilutions was fewer than 10 organisms per PCR. Intraocular sampling or corneal scraping was undertaken for all patients with suspected infectious endophthalmitis or keratitis (nonherpetic), respectively, between November 1999 and February 2001. PCRs were subsequently performed with 11 ocular samples. The amplified DNA was sequenced, and aligned against sequences in GenBank at the National Institutes of Health. The results were PCR positive for fungal primers for three corneal scrapings, one aqueous sample, and one vitreous sample; one of them was negative by culture. Molecular fungal identification was successful in all cases. Bacterial detection by PCR was positive for three aqueous samples and one vitreous sample; one of these was negative by culture. Amplification of ITS2/5.8S rDNA and molecular typing shows potential as a rapid technique for identifying fungi in ocular samples. PMID- 11474007 TI - Comparison of the BacT/Alert PF pediatric FAN blood culture bottle with the standard pediatric blood culture bottle, the Pedi-BacT. AB - The performance of the BacT/Alert PF (Organon-Teknika Corp., Durham, N.C.), a new nonvented pediatric FAN blood culture bottle, was compared to that of the original pediatric bottle, the Pedi-BacT, with matched aerobic cultures obtained from two separate facilities. A total of 244 clinically significant isolates were recovered from 4,015 compliant pairs. Among the positive cultures, 170 (70%) isolates were detected in both the BacT/Alert PF and the Pedi-BacT bottles, while 47 (19%) isolates were recovered in the BacT/Alert PF bottle only and 27 (11%) isolates were recovered in the Pedi-BacT bottle only. Although isolation of specific microorganisms was comparable for the two bottles, the total number of organisms recovered by the BacT/Alert PF was greater than that by the Pedi-BacT (P = 0.0272). In addition, more organisms were recovered by the BacT/Alert PF bottle from the blood of patients receiving antimicrobial therapy (P = 0.0180). Overall time to detection was similar for the two bottles; however, a significantly decreased mean time to detection was recorded for yeast from the BacT/Alert PF bottle (22.9 h; P = 0.0001) and staphylococci from the Pedi-BacT bottle (22.5 h; P = 0.0056). One false-negative culture and five false-positive cultures occurred with the Pedi-BacT bottle, compared to one false-positive culture with the BacT/Alert PF bottle. The BacT/Alert PF bottle is a reliable blood culture bottle for pediatric blood culture specimens and may offer improved recovery of microbes from patients on antimicrobial therapy. The use of the nonvented bottle will both facilitate bottle processing and decrease expenditures for materials due to the elimination of the venting needles required for the original vented bottles. PMID- 11474008 TI - Malaria rapid diagnostic devices: performance characteristics of the ParaSight F device determined in a multisite field study. AB - Microscopic detection of parasites has been the reference standard for malaria diagnosis for decades. However, difficulty in maintaining required technical skills and infrastructure has spurred the development of several nonmicroscopic malaria rapid diagnostic devices based on the detection of malaria parasite antigen in whole blood. The ParaSight F test is one such device. It detects the presence of Plasmodium falciparum-specific histidine-rich protein 2 by using an antigen-capture immunochromatographic strip format. The present study was conducted at outpatient malaria clinics in Iquitos, Peru, and Maesod, Thailand. Duplicate, blinded, expert microscopy was employed as the reference standard for evaluating device performance. Of 2,988 eligible patients, microscopy showed that 547 (18%) had P. falciparum, 658 (22%) had P. vivax, 2 (0.07%) had P. malariae, and 1,750 (59%) were negative for Plasmodium. Mixed infections (P. falciparum and P. vivax) were identified in 31 patients (1%). The overall sensitivity of ParaSight F for P. falciparum was 95%. When stratified by magnitude of parasitemia (no. of asexual parasites per microliter of whole blood), sensitivities were 83% (>0 to 500 parasites/microl), 87% (501 to 1,000/microl), 98% (1,001 to 5,000/microl), and 98% (>5,000/microl). Device specificity was 86%. PMID- 11474009 TI - Burkholderia cepacia complex infection in Italian patients with cystic fibrosis: prevalence, epidemiology, and genomovar status. AB - The prevalence, epidemiology, and genomovar status of Burkholderia cepacia complex strains recovered from Italian cystic fibrosis (CF) patients were investigated using genetic typing and species identification methods. Four CF treatment centers were examined: two in Sicily, one in central Italy, and one in northern Italy. B. cepacia complex bacteria were isolated from 59 out of 683 CF patients attending these centers (8.6%). For the two geographically related treatment centers in Sicily, there was a high incidence of infection caused by a single epidemic clone possessing the cblA gene and belonging to B. cepacia genomovar III, recA group III-A, closely related to the major North America United Kingdom clone, ET12; instability of the cblA sequence was also demonstrated for clonal isolates. In summary, of all the strains of B. cepacia encountered in the Italian CF population, the genomovar III, recA group III-A strains were the most prevalent and transmissible. However, patient-to-patient spread was also observed with several other genomovars, including strains of novel taxonomic status within the B. cepacia complex. A combination of genetic identification and molecular typing analysis is recommended to fully define specific risks posed by the genomovar status of strains within the B. cepacia complex. PMID- 11474010 TI - Serosubtypes and PorA types of Neisseria meningitidis serogroup B isolated in Brazil during 1997--1998: overview and implications for vaccine development. AB - Meningococcal disease caused by N. meningitidis serogroup B (MenB) has been endemic in Brazil since 1997. In this study, we determined the prevalence of serosubtypes of MenB isolated in 10 Brazilian states and the Federal District during 1997 and 1998 and investigated the extent of PorA VR sequence variation among the most prevalent serosubtypes to evaluate the possible use of an outer membrane vesicle (OMV)-, PorA-based vaccine to prevent meningococcal disease in Brazil. During this period, a total of 8,932 cases of meningococcal disease were reported. Only 42% (n = 3,751) of the reported cases were laboratory confirmed, and about 60% (n = 2,255) of those were identified as MenB. Among 1,297 MenB strains selected for this study, the most prevalent serosubtypes were P1.19,15 (66%), P1.7,1 (11%), and P1.7,16 (4%). PorA VR typing showed that 91% of the P1.19,15 strains analyzed had VR1 and VR2 sequences identical to those of the prototype strain. No sequence variation was detected among the 40 strains representing all isolated MenB P1.7,16 strains in the three southern states, where this serosubtype accounts for 75% of the serosubtypes identified. Similarly, all P1.7,1 strains were identified by PorA typing as P1.7-1,1. Although further improvements in the reporting of cases and collection of strains in Brazil are needed, our data suggest that a trivalent OMV-based vaccine prepared with PorA types P1.19,15, P1.7-1,1, and P1.7,16 may be appropriate to control serogroup B meningococcal disease in most of the Brazilian states. PMID- 11474011 TI - Development of conventional and real-time PCR assays for detection of Legionella DNA in respiratory specimens. AB - The development and validation of a PCR assay based on the use of new 16S ribosomal DNA (rDNA)-targeted primers to detect Legionella DNA in respiratory specimens are described. The assay was originally developed as conventional PCR followed by electrophoretic detection and was then adapted to Lightcycler format with SYBR Green I detection and melting curve analysis. The 73 Legionella pneumophila strains tested were amplified with both applications. In addition, 21 and 23 out of 27 other Legionella strains were found positive by conventional and real-time PCR assays, respectively, including the clinically important species L. micdadei, L. bozemaniae, and L. dumoffii. Two DNA purification methods were compared using artificially seeded clinical specimens: a standard organic extraction method and a commercial kit based on adsorption of DNA to silica particles. The detection limit of the assay varied from 2 CFU to >200,000 CFU per ml of clinical specimen, depending on the background sample (i.e., pooled sputa or BAL fluids) and the DNA purification method, the silica method achieving lower detection limits. Analysis of 77 clinical samples (66 bronchoalveolar lavage fluid and 11 sputum samples) by conventional PCR yielded results that were consistent with Legionella culture results. The melting curve analysis in the Lightcycler system readily detected the specific amplification products. However, run-to-run variations in the measured melting temperatures required normalization against the standard sample in each run. The results obtained with the clinical specimens were similar to those obtained with conventional PCR, but more samples are required to determine whether the system can be applied to routine screening of samples for the presence of Legionella DNA. PMID- 11474012 TI - Molecular cloning and sequencing of the circumsporozoite protein gene from Plasmodium falciparum strain FCC-1/HN and expression of the gene in Mycobacteria. AB - Mycobacterium bovis bacillus Calmette-Guerin (BCG) has been used as a live bacterial vaccine to immunize more than 2 billion people against tuberculosis. In an attempt to use this vaccine strain as a vehicle for protective antigens, the Plasmodium falciparum gene from strain FCC-1/HN encoding circumsporozoite protein (CSP) was amplified from the P. falciparum genome, sequenced, and expressed in M. bovis BCG under the control of an expression cassette carrying the promoter of heat shock protein 70 (HSP70) from Mycobacterium tuberculosis. The recombinant shuttle plasmid pBCG/CSP was introduced into mycobacteria by electroporation, and the recombinant mycobacteria harboring pBCG/CSP could be induced by heating to express CSP; the molecular mass of recombinant CSP was about 42 kDa. This report of expression of the almost-full-length P. falciparum CSP gene in BCG provides scientific evidence for the application of the HSP70 promoter in expressing a foreign gene in BCG and in development of BCG as a multivalent vectoral vaccine for malaria. PMID- 11474013 TI - Identification of Enterococcus, Streptococcus, and Staphylococcus by multivariate analysis of proton magnetic resonance spectroscopic data from plate cultures. AB - A new fingerprinting technique with the potential for rapid identification of bacteria was developed by combining proton magnetic resonance spectroscopy ((1)H MRS) with multivariate statistical analysis. This resulted in an objective identification strategy for common clinical isolates belonging to the bacterial species Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, and the Streptococcus milleri group. Duplicate cultures of 104 different isolates were examined one or more times using (1)H MRS. A total of 312 cultures were examined. An optimized classifier was developed using a bootstrapping process and a seven-group linear discriminant analysis to provide objective classification of the spectra. Identification of isolates was based on consistent high-probability classification of spectra from duplicate cultures and achieved 92% agreement with conventional methods of identification. Fewer than 1% of isolates were identified incorrectly. Identification of the remaining 7% of isolates was defined as indeterminate. PMID- 11474014 TI - Evaluation of laboratory testing methods for Chlamydia trachomatis infection in the era of nucleic acid amplification. AB - Diagnostic tests presently available for Chlamydia trachomatis have widely varying performance characteristics. To assess evolving laboratory testing practices since the introduction of nucleic acid amplification tests (NAAT), we surveyed laboratories in Washington State about their testing practices in 1998 and compared our findings to a similar survey conducted in 1995. Laboratory directors of 61 (87%) of 70 laboratories performing chlamydial tests in 1998 returned a survey. Between 1995 and 1998, 36 laboratories discontinued chlamydial testing, and the total number of laboratories performing tests in the state decreased from 92 to 70, a 24% decline. Of the 36 laboratories that discontinued testing, 25 (69%) had previously used rapid tests. While no laboratory routinely used NAAT in 1995, ligase chain reaction (LCR) was used in 23% of laboratories in 1998 and accounted for 113,624 (36%) of the 318,133 tests performed that year. Among the remaining 204,509 tests performed in 1998, other tests employed included DNA probe (29%), enzyme immunoassay (20%), culture (12%), direct fluorescent antibody assays (3%), and rapid tests (<1%). The majority (65%) of tests performed in 1998 using technologies other than LCR or culture were done in laboratories that did more than 10,000 tests. Cost and loss of revenue to laboratories were the most frequently cited reasons for not adopting NAAT. We conclude that in Washington State, NAAT have been rapidly adopted in larger laboratories, but most patients are still tested with much less sensitive technologies. Financial constraints represent the major barrier to more widespread use of DNA amplification tests. PMID- 11474015 TI - Evaluation of COBAS AMPLICOR (Roche): accuracy in detection of Chlamydia trachomatis and Neisseria gonorrhoeae by coamplification of endocervical specimens. AB - We evaluated further the accuracy of the COBAS AMPLICOR (Roche) (CA) PCR-based system in detection of Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical specimens. Endocervical specimens collected for any indication for testing for C. trachomatis and N. gonorrhoeae among a university hospital health system population were included. Testing for C. trachomatis was done by two PCR methods, CA and manual microwell AMPLICOR (Roche) (MWA), and by culture; testing for N. gonorrhoeae was done by CA and culture. Discrepancy resolution was performed. Reproducibility testing and hands-on labor time measurements for CA were done. Among 654 C. trachomatis samples, the prevalence of true positivity was 9.2%, and among the 618 N. gonorrhoeae samples, the prevalence of true positivity was 4.4%. For detection of C. trachomatis, the sensitivity, specificity, and negative and positive predictive values were, respectively, as follows for each test: CA, 93.3, 99.7, 99.3, and 96.4%; MWA, 91.7, 99.7, 99.2, and 96.5%; and culture, 65.0, 100, 96.6, and 100%. For detection of N. gonorrhoeae those values were as follows: CA, 96.3, 100, 99.8, and 100%; and culture, 92.6, 100, 99.7, and 100%. Hands-on labor time for each clinical result was estimated to be at 7.5 min. The prevalence of inhibitory specimens was 3.5%, including two positive C. trachomatis samples which would have been missed otherwise. The direct cost of each clinical result with CA was estimated to be $9.09. Our methods include a diverse range of indications for testing among women, using endocervical swabbing samples, 2 M sucrose phosphate transport medium, and discrepancy resolution for comparison. Under our test conditions, the CA system is an accurate, rapid, and cost- and labor-efficient method for detection of C. trachomatis and N. gonorrhoeae. PMID- 11474016 TI - Evaluation of BACTEC MYCO/F Lytic medium for recovery of mycobacteria, fungi, and bacteria from blood. AB - MYCO/F Lytic medium (MFL), a liquid medium developed for use with the BACTEC 9240 blood culture system, was compared to the Isolator system (IS) for the recovery of fungi and to the BACTEC 13A medium for the recovery of mycobacteria. Recovery of bacteria was compared to routine BACTEC Plus Aerobic/F (AF) blood cultures. Microbial growth was detected in 203 (17%) of 1,166 blood cultures. Fifty-seven specimens were positive for fungi: 35 were positive with both IS and MFL; six were positive with IS only (three Candida albicans, one Histoplasma capsulatum, one Candida glabrata, and one Fusarium species isolate); three were positive with AF only (two C. albicans and one Candida parapsilosis isolate); and 13 were positive with MFL only (five C. glabrata, three C. albicans, two Candida krusei, two Candida tropicalis, and one C. parapsilosis isolate; P > 0.05 versus IS). Eighteen of 19 blood cultures positive for H. capsulatum grew in both IS and MFL, although the time to detection for MFL was greater. The mean time to detection for all fungi was 8.15 days for IS and 12.07 days for MFL. Seven hundred forty specimens were also cultured for mycobacteria with MFL and 13A. Forty-four grew mycobacteria; 38 were positive with both 13A and MFL; and 16 were positive with MFL only. Mycobacterium avium was recovered from 41 specimens; 36 were positive for both systems and 5 were positive for MFL alone. MFL was also compared to the AF bottle for the same 740 specimens. MFL and AF both detected 34 of the 40 clinically significant bacteria, while IS detected only 15 of 40. In summary, MFL is an excellent medium for the recovery of fungi, mycobacteria, and bacteria; however, the time to detection of H. capsulatum is increased. PMID- 11474017 TI - Automated multiplex assay system for simultaneous detection of hepatitis B virus DNA, hepatitis C virus RNA, and human immunodeficiency virus type 1 RNA. AB - We have developed an automated multiplex system for simultaneously screening hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1) in blood donations. The assay, designated AMPLINAT MPX HBV/HCV/HIV-1 Test (AMPLINAT MPX), consists of virus extraction and target sequence-specific probe capture on specimen preparation workstation GT-X (Roche Diagnostics K.K., Tokyo, Japan) and amplification and detection by TaqMan PCR on the ABI PRISM 7700 Analyzer (Perkin-Elmer Applied Biosystems, Foster City, Calif.). An internal control (IC) is incorporated in the assay to monitor the extraction, target amplification, and detection processes. The assay yields qualitative results without discrimination of the three targets. Detection limits (95% confidence interval) are 22 to 60 copies/ml for HBV, 61 to 112 IU/ml for HCV, and 33 to 66 copies/ml for HIV-1, using a specimen input volume of 0.2 ml. The AMPLINAT MPX assay detects a broad range of genotypes or subtypes for all three viruses and has a specificity of 99.6% for all three viruses with seronegative specimens. In an evaluation of seroconversion panels, the AMPLINAT MPX assay detects HBV infection an average of 24 days before the detection of HBsAg by enzyme immunoassay. HCV RNA was detected an average of 31 days before HCV antibody. HIV-1 RNA was detected an average of 14 days before HIV-1 antibody and an average of 9 days before p24 antigen. The Japanese Red Cross has been evaluating the AMPLINAT MPX system since October 1999. The clinical performance indicates that the AMPLINAT MPX system is robust, sensitive, and reproducible, with a high percentage of valid assay runs (96.8%), a low false-positive rate (0.34%), and a low IC failure rate (0.24%). PMID- 11474018 TI - Detection of cervical infections in colposcopy clinic patients. AB - The purpose of this study was to determine if Neisseria gonorrhoeae; Chlamydia trachomatis; herpes simplex virus; cytomegalovirus; Epstein-Barr virus; human herpesviruses 6, 7, and 8; or adeno-associated virus influenced the production of cervical intraepithelial neoplasia. Two hundred thirty-one cervical smear samples were tested for the presence of the organisms by PCR. In addition, human papillomavirus types in the samples were determined by PCR and classified into cancer risk types of high, moderate, and low. There was no link with cervical intraepithelial neoplasia status and detection of herpes simplex virus, cytomegalovirus, Epstein-Barr virus, human herpesviruses 6 and 8, gonorrhea, or chlamydia. However, high-grade cervical intraepithelial neoplasia was found more frequently with mixed infection by moderate-risk human papillomavirus types and human herpesvirus 7 than with these papillomavirus types alone. The presence of human herpesvirus 7 may increase the oncogenic potential of moderate-risk human papillomavirus types. PMID- 11474019 TI - Cyclospora cayetanensis in three populations at risk in Guatemala. AB - In 1996 and 1997, outbreaks of Cyclospora cayetanensis in North America were linked to Guatemalan raspberries. From April 1999 to April 2000, we undertook a survey for C. cayetanensis in raspberry farm workers, malnourished children, and human immunodeficiency virus and AIDS patients in Guatemala. Stool samples were analyzed using ethylacetate-formalin concentration, wet preparation, modified acid-fast staining method, and epifluorescence. Oocysts were found in 1.5% of the subjects, none of whom were raspberry farm workers. PMID- 11474020 TI - Characterization of Borrelia burgdorferi isolated from erythema migrans lesions: interrelationship of three molecular typing methods. AB - Genetic diversity among Borrelia burgdorferi isolates recovered from the skin of Lyme disease patients was assessed by ribosomal DNA (rDNA) spacer restriction fragment length polymorphism analysis, genomic restriction site polymorphism analysis, and plasmid content analysis. There was a significant association between the three rDNA spacer types, the six pulsed-field gel types, and plasmid content (P < 0.001). The association between distinct chromosomal and plasmid markers implies a clonal origin for each genotype. PMID- 11474021 TI - Efficacy of a swab transport system in maintaining viability of Neisseria gonorrhoeae and Streptococcus pneumoniae. AB - The efficacy of swab transport systems in maintaining viability of Neisseria gonorrhoeae and Streptococcus pneumoniae is crucial both for establishing definitive diagnosis and for monitoring emerging resistance. We tested the efficacy of a newly modified Amies charcoal swab transport system, the StarSwab SP131X (Starplex Scientific, Inc., Etobicoke, Ontario, Canada), by using a combined total of 31 clinical and American Type Culture Collection stock reference strains of N. gonorrhoeae and S. pneumoniae in 46 suspensions of concentrations ranging from 10(5) to 10(8) CFU/ml. Triplicate swabs per strain held at room temperature for 0, 24, and 48 h were plated without prior vortexing, and their growths were graded. All 31 strains were viable at 0 and 24 h. Gonococcal viability at 48 h varied considerably, even among strains with comparable inoculum sizes, suggesting that viability might be strain dependent and confirming the different structural and growth profiles of gonococcal strains. S. pneumoniae strains showed consistent viability, with all strains recovered at all holding periods. This study demonstrates that the StarSwab SP131X is capable of maintaining the viability of N. gonorrhoeae and S. pneumoniae for at least 24 and 48 h, respectively, and reinforces the need for adequate sampling and for timely processing of specimens to maintain optimum performance. PMID- 11474022 TI - Correlation between genotype and phenotypic categorization of staphylococci based on methicillin susceptibility and resistance. AB - Positive correlation between methicillin and oxacillin susceptibility test results and the detection of the mecA gene was observed for Staphylococcus aureus, S. epidermidis, and S. haemolyticus as well as among mecA(+) strains of other species of coagulase-negative staphylococci (CNS). However, at least 50% of the mecA-negative strains of these other species of CNS were falsely classified as methicillin and oxacillin resistant. PMID- 11474023 TI - Evaluation of VITEK 2 rapid identification and susceptibility testing system against gram-negative clinical isolates. AB - A total of 281 strains of miscellaneous members of the family Enterobacteriaceae, Pseudomonas aeruginosa, and other gram-negative bacteria were evaluated by use of identification tests with the VITEK 2 system (bioMerieux) and an API identification system (bioMerieux). A total of 237 (95%) strains were correctly identified to the species level. Only six (2.1%) strains were misidentified, and eight (2.8%) strains were not identified. Among 14 strains with discrepant identifications, 8 (57.1%) strains were nonfermenters. The susceptibilities of 228 strains to 11 antibiotics including amikacin, netilmicin, tobramycin, gentamicin, ciprofloxacin, imipenem, meropenem, ceftazidime, cefepime, piperacillin, and piperacillin in combination with tazobactam were tested with the VITEK 2 AST-No. 12 card and by the broth microdilution (MB) method, according to NCCLS guidelines, as a reference. For the 2,508 organism-antibiotic combinations, the rates at which duplicate MICs correlated within +/-1 dilution ranged from 84.2 to 95.6%. Only 13 (0.5%) and 10 (0.4%) of the susceptibility tests gave major errors (resistant with the VITEK 2 system but sensitive by the MB method) and very major errors (sensitive with the VITEK 2 system but resistant by the MB method), respectively. Both VITEK 2 ID-GNB (an identification system) and VITEK 2 AST-No. 12 (a susceptibility testing system) card systems gave rapid, reliable, and highly reproducible results. PMID- 11474024 TI - Detection of antibodies to U.S. isolates of avian pneumovirus by a recombinant nucleocapsid protein-based sandwich enzyme-linked immunosorbent assay. AB - The nucleocapsid (N) protein of subgroup C (United States-specific) avian pneumovirus (APV/US) was expressed in Escherichia coli, and antibodies to the recombinant N protein were shown to specifically recognize the approximately 47 kDa N protein of APV/US by Western immunoblot analysis. The recombinant APV/US N protein was used in a sandwich-capture enzyme-linked immunosorbent assay (ELISA), and the resulting assay was found to be more sensitive and specific than the routine indirect ELISA for the detection of APV/US antibodies in turkey sera. PMID- 11474025 TI - Candida species exhibit differential in vitro hemolytic activities. AB - A total of 80 Candida isolates representing 14 species were examined for their respective responses to an in vitro hemolytic test. A modification of a previously described plate assay system where the yeasts are incubated on glucose (3%)-enriched sheep blood agar in a carbon dioxide (5%)-rich environment for 48 h was used to evaluate the hemolytic activity. A group of eight Candida species which included Candida albicans (15 isolates), C. dubliniensis (2), C. kefyr (2), C. krusei (4), C. zeylanoides (1), C. glabrata (34), C. tropicalis (5), and C. lusitaniae (2) demonstrated both alpha and beta hemolysis at 48 h postinoculation. Only alpha hemolysis was detectable in four Candida species, viz., C. famata (3), C. guilliermondii (4), C. rugosa (1), and C. utilis (1), while C. parapsilosis (5) and C. pelliculosa (1) failed to demonstrate any hemolytic activity after incubation for 48 h or longer. This is the first study to demonstrate the variable expression profiles of hemolysins by different Candida species. PMID- 11474026 TI - Identification of the hexon region of an adenovirus involved in a new outbreak of keratoconjunctivitis. AB - We tested 15 adenovirus (Ad)-positive patients involved in a case of nosocomial spread of keratoconjunctivitis. A neutralization test, PCR-restriction fragment length polymorphism analysis, and sequencing of the hypervariable regions of the hexons were performed in order to identify the type of Ad involved. The serotype of the Ad was not identical to any published Ad sequence by either method. PMID- 11474027 TI - Immunofluorescent detection of intraerythrocytic Bartonella henselae in naturally infected cats. AB - To determine the presence of Bartonella henselae bacteremia in six cats, we compared isolation using blood culture with direct immunofluorescence on blood smears. Three cats that were positive by blood culture were also positive by direct immunofluorescence, and laser confocal microscopy confirmed the intraerythrocytic location of B. henselae. PMID- 11474028 TI - Atypical, fljB-negative Salmonella enterica subsp. enterica strain of serovar 4,5,12:i:- appears to be a monophasic variant of serovar Typhimurium. AB - An fljB-negative, multidrug-resistant Salmonella enterica serovar 4,5,12:i:- phage type DT U302 strain (resistant to ampicillin, chloramphenicol, sulfonamide, gentamicin, streptomycin, tetracycline, and sulfamethoxazole-trimethoprim) emerged and spread in Spain in 1997. Sequences specific for Salmonella serovar Typhimurium and phage type DT 104 and U302 were present in this atypical Salmonella strain, suggesting that it is a monophasic Salmonella serovar Typhimurium variant. PMID- 11474029 TI - Simplified microneutralization test for serotyping adenovirus isolates. AB - A simplified microneutralization procedure is described that uses an empirically determined virus challenge dose, a single dilution of antiserum, and observation of cytopathic effect to determine the adenovirus serotype. The simplified test has faster turnaround time and was 96% concordant with a confirmatory test using serial dilutions of type-specific sera. This method will find utility in high volume serotyping work. PMID- 11474030 TI - Mutations in the rpoB gene of multidrug-resistant Mycobacterium tuberculosis clinical isolates from India. AB - Mutations in the 81-bp rifampin resistance-determining region (RRDR) of the rpoB gene were analyzed by DNA sequencing of 50 Mycobacterium tuberculosis clinical isolates (44 resistant and 6 sensitive) from various parts of India. Fifty-three mutations of 18 different kinds, 17 point mutations and one deletion, were observed in 43 of 44 resistant isolates. Three novel mutations and three new alleles within the RRDR, along with two novel mutations outside the RRDR, are reported in this study. PMID- 11474031 TI - Possible approach for serodiagnosis of ascariasis by evaluation of immunoglobulin G4 response using Ascaris lumbricoides somatic antigen. AB - Somatic antigen of Ascaris lumbricoides was purified to homogeneity (molecular mass, 34 kDa) by ammonium sulfate fractionation and successive chromatographic procedures, namely, gel permeation, ion exchange, and high-performance gel permeation liquid chromatographies. The antigen showed strong binding with immunoglobulin G (IgG) in Ascaris-infested patients and was cross-reactive with IgE and IgG in patients infested with other nematodes. It reacted specifically with IgG4 (P < 0.001) in 63 Ascaris-infested patients, which represented 65% of the total IgG response, though cross-reactivity with IgG1, IgG2, and IgG3 subclasses was observed, indicating the unique specificity of this test system and its potential utility in the serodiagnosis of ascariasis. PMID- 11474032 TI - Genetic diversity of Pneumocystis carinii isolated from human immunodeficiency virus-positive patients in Turin, Italy. AB - By DNA sequence analysis we identified two new strain types and five novel sporadic variations among 25 isolates of Pneumocystis carinii f. sp. hominis obtained from 19 human immunodeficiency virus-positive patients. Of these, 13 were infected with a single strain and 6 were coinfected. Fifteen different combination types were identified among the 18 strains for which complete molecular typing was accomplished. PMID- 11474033 TI - Serological differentiation of experimentally induced Candida dubliniensis and Candida albicans infections. AB - Using a rabbit model of systemic infection, we show that it is possible to differentiate infections caused by Candida dubliniensis and other Candida species by detecting the antibody response mounted by the infected animals. These results confirm our previous observation in a patient with C. dubliniensis candidemia and suggest that detection of C. dubliniensis-specific antibodies is useful in the diagnosis of invasive candidiasis caused by this yeast. PMID- 11474034 TI - Automated ribotyping using different enzymes to improve discrimination of Listeria monocytogenes isolates, with a particular focus on serotype 4b strains. AB - To develop improved automated subtyping approaches for Listeria monocytogenes, we characterized the discriminatory power of different restriction enzymes for ribotyping. When 15 different restriction enzymes were used for automated ribotyping of 16 selected L. monocytogenes isolates, the restriction enzymes EcoRI, PvuII, and XhoI showed high discriminatory ability (Simpson's index of discrimination > 0.900) and produced complete and reproducible restriction cut patterns. These three enzymes were thus evaluated for their ability to differentiate among isolates representing the two major serotype 4b epidemic clones, those having ribotype reference pattern DUP-1038 (51 isolates) and those having pattern DUP-1042 (20 isolates). Among these isolates, PvuII provided the highest discrimination for a single enzyme (nine different subtypes; index of discrimination = 0.518). A combination of PvuII and XhoI showed the highest discriminatory ability (index of discrimination = 0.590) for these isolates. A group of 44 DUP-1038 isolates and a group of 12 DUP-1042 isolates were identical to each other even when the combined data for all three enzymes were used. We conclude that automated ribotyping using different enzymes allows improved discrimination of L. monocytogenes isolates, including epidemic serotype 4b strains. We furthermore confirm that most of the isolates representing the genotypes linked to the two major epidemic L. monocytogenes clonal groups form two genetically homogeneous groups. PMID- 11474035 TI - Rapid pulsed-field gel electrophoresis method for group B streptococcus isolates. AB - We developed a rapid pulsed-field gel electrophoresis (PFGE) method that required 3 days to complete, an improvement over the standard method that required as many as 8 days. The accuracy and reproducibility of the rapid method were verified by analysis of DNA band sizes of our control group B streptococcus isolate. The rapid method was superior to the standard method, providing more precise molecular sizing and gels of higher image quality. The reproducibility of rapid PFGE substantiated its value and continued use. PMID- 11474036 TI - Cariogenic actinomyces identified with a beta-glucosidase-dependent green color reaction to Gardenia jasminoides extract. AB - The oral bacteria Actinomyces naeslundii and Actinomyces viscosus are known to contribute to the initiation and progression of human dental caries, especially root caries. We report that both A. naeslundii and A. viscosus react with a component in the Gardenia jasminoides extract to produce a distinct green product. This green color reaction was found to be dependent on the bacterial beta-glucosidase. The reaction is specific for cariogenic actinomyces, and it can detect as few as 10(4) cells of A. naeslundii and A. viscosus per ml. PMID- 11474037 TI - Waldenstrom's disease complicated by recurrent meningococcal arthritis. AB - Meningococcal arthritis is rare. We report a patient in whom a first episode of meningococcal arthritis revealed Waldenstrom's disease and who experienced a second episode of meningococcal arthritis 8 years later. We suggest that an impaired immune response secondary to Waldenstrom's disease favored the recurrence of meningococcal arthritis. PMID- 11474038 TI - Suicide and gun control. PMID- 11474040 TI - Best practices: dissemination of guidelines for the treatment of major depression in a managed behavioral health care network. PMID- 11474041 TI - Consumer & family information: premenstrual dysphoric disorder. PMID- 11474042 TI - Practical psychotherapy: how practice guidelines can rescue psychotherapy in public systems. PMID- 11474043 TI - Economic grand rounds: inpatient and partial hospital care under medicare. PMID- 11474044 TI - Datapoints: trends in psychiatric practice, 1988-1998: III. Activities and work settings. PMID- 11474045 TI - Alcohol & drug abuse: a preliminary investigation of cocaine craving among persons with and without schizophrenia. PMID- 11474046 TI - Psychopharmacology: rational and irrational polypharmacy. PMID- 11474047 TI - Personal accounts: memoirs of a compulsive firesetter. PMID- 11474048 TI - Some perspectives on deinstitutionalization. AB - The authors discuss what can be learned from our experience with deinstitutionalization. The deinstitutionalization of mentally ill persons has three components: the release of these individuals from hospitals into the community, their diversion from hospital admission, and the development of alternative community services. The greatest problems have been in creating adequate and accessible community resources. Where community services have been available and comprehensive, most persons with severe mental illness have significantly benefited. On the other hand, there have been unintended consequences of deinstitutionalization-a new generation of uninstitutionalized persons who have severe mental illness, who are homeless, or who have been criminalized and who present significant challenges to service systems. Among the lessons learned from deinstitutionalization are that successful deinstitutionalization involves more than simply changing the locus of care; that service planning must be tailored to the needs of each individual; that hospital care must be available for those who need it; that services must be culturally relevant; that severely mentally ill persons must be involved in their service planning; that service systems must not be restricted by preconceived ideology; and that continuity of care must be achieved. PMID- 11474049 TI - A critical review of internet information about depression. AB - OBJECTIVE: The source and quality of information about depression available on the Internet were examined. METHODS: Online searches using the phrase "depression and treatment" were conducted on ten major Internet search engines. The first 20 sites generated by each engine were examined. The Web sites and the individual Web pages they contained were categorized by source, target audience, and affiliation with either a for-profit or a not-for-profit organization. Each site was rated by whether it mentioned the nine symptoms and five major criteria of a major depressive episode and whether it made any of three basic treatment recommendations. Ratings were compared to determine whether treatment differences between the sites could be attributed to the site's source. RESULTS: The search generated a total of 178 active sites. Overall, the quality of information was poor. Only half of the sites mentioned any DSM-IV diagnostic symptom or criteria in their descriptions of depression, and only a quarter listed 11 to 14 symptoms or criteria. Almost half of the sites made no mention of medications, psychotherapy, or professional consultation as suggested treatments for depression. Almost two-thirds of the pages were found on for-profit sites. Documents from these sites scored lower on diagnostic accuracy and treatment recommendations than those from not-for-profit sites. CONCLUSIONS: The quality of information on the Internet produced by the search was quite low. For-profit Web sites appeared much more frequently than not-for-profit sites among the first 20 sites generated by each search engine, and they contained poorer information. PMID- 11474050 TI - Long-stay patients in state psychiatric hospitals at the end of the 20th century. AB - OBJECTIVES: The purpose of this study was to assess the characteristics of long stay patients in contemporary state psychiatric hospitals and to identify factors representing possible barriers to alternative placements for these patients. METHODS: All patients in inpatient units of the Massachusetts Department of Mental Health who had been hospitalized for at least three years as of April 1, 1999, were assessed by their treatment teams with a standardized data collection instrument. Domains assessed included medical problems, need for nursing care, psychiatric diagnosis, and history of problematic behaviors. RESULTS: The 330 individuals identified as long-stay patients had an array of medical problems and nursing care needs that likely would have been manageable in other long-term-care settings. A total of 276 patients had at least one significant medical problem. However, some patients exhibited behavioral problems that might have complicated such placements, especially when behavioral problems co-occurred with the need for medical supervision. A total of 228 patients had exhibited a significant problematic behavior in the previous 30 days. CONCLUSIONS: Although the number of long-stay patients in state psychiatric hospitals declined dramatically during the second half of the 20th century, a small group of patients still requires care in this setting. State psychiatric hospitals continue to occupy a significant niche in the mental health system. PMID- 11474051 TI - Firearms risk management in psychiatric care. AB - OBJECTIVE: This study sought to assess the effectiveness of a firearms risk management program. METHODS: A firearms risk management program, which included multidisciplinary assessment, treatment, and discharge planning, was developed and implemented among 46 civilly committed psychiatric inpatients at the Twin Valley Psychiatric System, a behavioral health organization of the Ohio Department of Mental Health. RESULTS: The research sample comprised mainly men who had personality disorders and histories of substance abuse and who had expressed an intent to use a firearm to commit suicide. On discharge, none of the patients had access to a firearm. Of the 16 patients who were hospitalized during the next 24 months, only five were noted to have threatened to harm themselves or others with a firearm or to have access to a firearm. CONCLUSIONS: Multidisciplinary and focused assessment, treatment, and discharge planning can be effective in neutralizing the risk of firearms use among psychiatric patients. PMID- 11474052 TI - A national survey of care for persons with co-occurring mental and substance use disorders. AB - OBJECTIVE: The delivery of appropriate treatment to persons who have mental and substance use disorders is of increasing concern to clinicians, administrators, and policy makers. This study sought to describe use of appropriate mental health and comprehensive substance abuse care among adults in the United States with probable co-occurring disorders. METHODS: Data from the Healthcare for Communities survey, which is based on a national household sample studied in 1997 and 1998, were used to identify individuals who had probable co-occurring mental and substance use disorders. The sociodemographic and clinical characteristics of these individuals and their use of services were recorded. Logistic regression analysis was used to identify variables associated with receipt of mental health and substance abuse treatment and with receipt of appropriate treatment. RESULTS: Estimates for the U.S. adult population based on the weighted survey data indicated that 3 percent of the population had co-occurring disorders. Seventy two percent did not receive any specialty mental health or substance abuse treatment in the previous 12 months; only 8 percent received both specialty mental health care and specialty substance abuse treatment. Only 23 percent received appropriate mental health care, and 9 percent received supplemental substance abuse treatment. Perceived need for treatment was strongly associated with receipt of any mental health care and with receipt of appropriate care. CONCLUSIONS: Despite the availability of effective treatments, most individuals who had co-occurring mental health and substance use problems were not receiving effective treatment. Efforts to improve the care provided to persons who have co occurring disorders should focus on strategies that increase the delivery of effective treatment. PMID- 11474053 TI - Prevalence of loss and complicated grief among psychiatric outpatients. AB - OBJECTIVE: This study examined the prevalence of significant loss through the death of another person as well as complicated grief among patients at two psychiatric outpatient clinics for one year. METHODS: A total of 729 patients were interviewed about significant losses through death during their lives. Standard questionnaires were used to classify 235 patients who had experienced such losses into three groups: those who had minimal disturbance, those who had moderate complicated grief, and those who had severe complicated grief. Multivariate and univariate analyses of variance were used to test for differences in loss-specific variables (for example, pathological grief) and variables that were not specific to loss (for example, depression) among the three groups. RESULTS: More than half of the 729 patients reported that they had experienced one or more significant losses through death. About a third of all patients who came to the clinics met the criteria for either moderate or severe complicated grief. The average time since the loss was about ten years, indicating that these patients had long-term complicated grief. Significant differences in loss-specific variables and variables that were not specific to loss were detected among the three groups. Patients who had severe complicated grief scored higher than patients in the other two groups on both types of variables. Patients with moderate complicated grief had higher scores than those with minimal disturbance. CONCLUSIONS: Clinicians should routinely assess outpatients for loss and complicated grief and should consider addressing loss and complicated grief in treatment. Rather than a single classification of complicated grief, different levels should be considered. PMID- 11474054 TI - Vocational outcomes among formerly homeless persons with severe mental illness in the access program. AB - OBJECTIVE: This study examined the vocational outcomes of 4,778 formerly homeless individuals with severe mental illness who were enrolled in the Access to Community Care and Effective Services and Support (ACCESS) program, a multisite demonstration project designed to provide services to this population. METHODS: Participants were interviewed at the time of enrollment and again three months and 12 months later by trained researchers who were not part of the treatment team to determine their employment status. At 12 months, participants were also asked about the types of services they had received during the past 60 days. Multiple logistic regression analysis was used to predict employment at 12 months. RESULTS: ACCESS participants reported receiving relatively few job related services. Nonetheless, modest but significant increases occurred between baseline and three months and between three months and 12 months in the total proportion of participants who were employed and who were employed full-time and in hourly earnings and estimated monthly earnings. The number of hours worked per week increased significantly between three months and 12 months. When the analysis controlled for site, study condition (whether the ACCESS site received or did not receive extra funds to improve service integration), minority status, addiction treatment, and mental health treatment, participants who were employed at 12 months were more likely to have received job training and job placement services. CONCLUSIONS: Programs that work with homeless mentally ill persons may better serve their clients by placing as great an emphasis on providing employment services as on providing housing and clinical treatment. PMID- 11474055 TI - Prescription of psychotropic medications to youths in office-based practice. AB - OBJECTIVE: This study sought to determine sociodemographic characteristics of treatment of children and adolescents for whom psychotropic medications are prescribed and to describe the clinical management approaches associated with the prescription of each major class of psychotropic medication in office-based medical practices in the United States. METHODS: Data for a four-year period (1992-1996) were drawn from the National Ambulatory Medical Care Survey, a nationally representative survey of office-based medical practices, to determine prescribing patterns, patients' sociodemographic characteristics, and clinical management approaches associated with visits during which psychotropic medications were prescribed to patients aged 19 years or under. RESULTS: Psychotropic medications were prescribed during 2.2 percent of all visits. A majority of the prescriptions for psychotropic medications (84.8 percent) were provided by general practitioners or pediatricians. For the visits during which a psychotropic medication was prescribed, stimulants were the most commonly prescribed (53.9 percent of such visits), but prescription of other classes of medications was not uncommon: antidepressants (30 percent), anxiolytics (7.2 percent), antipsychotics (7.2 percent), and mood stabilizers (12.7 percent). Significant differences were observed in the prescription of each class of medication by sex, race, and payment source. CONCLUSIONS: General practitioners and pediatricians have a role in the office-based treatment of youths with psychotropic medications. PMID- 11474056 TI - Reliability of the services assessment for children and adolescents. AB - OBJECTIVE: This study examined the test-retest reliability of a new instrument, the Services Assessment for Children and Adolescents (SACA), for children's use of mental health services. METHODS: A cross-sectional survey was undertaken at two sites. The St. Louis site used a volunteer sample recruited from mental health clinics and local schools. The Ventura County, California, site used a double-blind, community-based sample seeded with cases of service-using children. Participating families completed the SACA and were retested within four to 14 days. The reliability of service use items was calculated with use of the kappa statistic. RESULTS: The SACA- Parent Version had excellent test-retest reliability for both lifetime service use and previous 12-month use. The SACA also had good to excellent reliability when administered to children aged 11 and older for lifetime and 12-month use. Reliability figures for children aged nine and ten years were considerably lower for lifetime and 12-month use. The younger children's responses suggested that they were confused about some questions. CONCLUSIONS: This study demonstrates that parents and older children can reliably report use of mental health services by using the SACA. The SACA can be used to collect currently unavailable information about use of mental health services. PMID- 11474057 TI - An analysis of reported deaths following electroconvulsive therapy in Texas, 1993 1998. AB - Since 1993, Texas law has required that all deaths that occur within 14 days of electroconvulsive therapy (ECT) be reported to the Texas Department of Mental Health and Mental Retardation. This study attempted to differentiate deaths that may have been due to ECT or the associated anesthesia from those due to other causes. Among more than 8,000 patients who received 49,048 ECT treatments between 1993 and 1998, a total of 30 deaths were reported to the mental health department between 1993 and 1998. Only one death, which occurred on the same day as the ECT, could be specifically linked to the associated anesthesia. An additional four deaths could plausibly have been associated with the anesthesia, for which the calculated mortality rate is between two and ten per 100,000, but probably not with the stimulus of the ECT or seizure. The mortality rate associated with ECT (less than two per 100,000 treatments) in Texas is extremely low. PMID- 11474058 TI - Validation of the brief instrumental functioning scale in a homeless population. AB - This report describes the psychometric properties of the six-item Brief Instrumental Functioning Scale (BIFS), a self-report questionnaire for measuring instrumental functioning in community populations, and the results of a study to validate the scale among homeless persons. The participants in the study were 1,509 homeless persons, of whom 1,077, or 71 percent, were seriously mentally ill, substance dependent, or both. Confirmatory factor analysis provided evidence for the unidimensionality of the items in the scale. The BIFS had high internal consistency, respectable stability, and reasonable convergent validity. The BIFS is useful for community populations when self-report data on functioning are required. Additional research is needed to develop and validate comparable scales for assessing other domains of functioning. PMID- 11474059 TI - Factors associated with readmission to a psychiatric facility. AB - This study examined patient-related factors that were associated with readmission to a tertiary care psychiatric hospital in Canada. The charts of a random sample of 200 patients were reviewed from an index discharge date in 1991 through subsequent rehospitalizations over the next three years. Eighty-eight patients (44 percent) were readmitted at least once. The only variable that significantly differentiated patients who were readmitted from those who were not was a history of admission. System variables or factors that are not patient related, such as staff attitudes and perceptions, may contribute to readmission and thus may warrant further exploration. PMID- 11474061 TI - Confusion about outpatient commitment. PMID- 11474062 TI - Who needs trauma initiatives? South Carolina! PMID- 11474063 TI - Smoking and panic disorder. PMID- 11474064 TI - Fluvoxamine and aggression in mental retardation. PMID- 11474065 TI - Alice James and endometriosis. PMID- 11474066 TI - Segregation of human neural stem cells in the developing primate forebrain. AB - Many central nervous system regions at all stages of life contain neural stem cells (NSCs). We explored how these disparate NSC pools might emerge. A traceable clone of human NSCs was implanted intraventricularly to allow its integration into cerebral germinal zones of Old World monkey fetuses. The NSCs distributed into two subpopulations: One contributed to corticogenesis by migrating along radial glia to temporally appropriate layers of the cortical plate and differentiating into lamina-appropriate neurons or glia; the other remained undifferentiated and contributed to a secondary germinal zone (the subventricular zone) with occasional members interspersed throughout brain parenchyma. An early neurogenetic program allocates the progeny of NSCs either immediately for organogenesis or to undifferentiated pools for later use in the "postdevelopmental" brain. PMID- 11474067 TI - Global analysis of protein activities using proteome chips. AB - To facilitate studies of the yeast proteome, we cloned 5800 open reading frames and overexpressed and purified their corresponding proteins. The proteins were printed onto slides at high spatial density to form a yeast proteome microarray and screened for their ability to interact with proteins and phospholipids. We identified many new calmodulin- and phospholipid-interacting proteins; a common potential binding motif was identified for many of the calmodulin-binding proteins. Thus, microarrays of an entire eukaryotic proteome can be prepared and screened for diverse biochemical activities. The microarrays can also be used to screen protein-drug interactions and to detect posttranslational modifications. PMID- 11474068 TI - Resistance to an herbivore through engineered cyanogenic glucoside synthesis. AB - The entire pathway for synthesis of the tyrosine-derived cyanogenic glucoside dhurrin has been transferred from Sorghum bicolor to Arabidopsis thaliana. Here, we document that genetically engineered plants are able to synthesize and store large amounts of new natural products. The presence of dhurrin in the transgenic A. thaliana plants confers resistance to the flea beetle Phyllotreta nemorum, which is a natural pest of other members of the crucifer group, demonstrating the potential utility of cyanogenic glucosides in plant defense. PMID- 11474069 TI - The Y chromosome and the replacement hypothesis. PMID- 11474070 TI - The biomedical research bottleneck. PMID- 11474072 TI - Global warming. World starts taming the greenhouse. PMID- 11474071 TI - High-energy physics. Plans for next big collider reach critical mass at Snowmass. PMID- 11474073 TI - Genome research: map of the human genome 3.0. PMID- 11474075 TI - New facilities. Funding backlog at NSF sets off free-for-all. PMID- 11474074 TI - Genome research. DNA sequencers to go bananas? PMID- 11474076 TI - Clinical research. Shutdown at Hopkins sparks a debate. PMID- 11474077 TI - Houston flood. Research toll is heavy in time and money. PMID- 11474078 TI - Animal behavior. New data reveal the sisterhood of lions. PMID- 11474079 TI - Ecology. Galapagos takes aim at alien invaders. PMID- 11474080 TI - North American Paleontological convention meeting. Fossils with lessons for conservation biology. PMID- 11474081 TI - Genetics. Can SNPs deliver on susceptibility genes? PMID- 11474082 TI - Genetics. SNP-ing drugs to size. PMID- 11474083 TI - Science competitions. Top young problem solvers vie for quiet glory. PMID- 11474084 TI - Science competitions. IMO's golden boy makes perfection look easy. PMID- 11474085 TI - Science competitions. Science olympiads offer a variety of arenas for overachievers. PMID- 11474086 TI - Animal research. Reliability of protocol reviews for animal research. PMID- 11474087 TI - Statistical thermodynamics. Taking a walk on a landscape. PMID- 11474088 TI - Cell biology. Surviving starvation. PMID- 11474089 TI - Volcanology. Predicting volcanic eruptions. PMID- 11474090 TI - Structural biology. Actin' up. PMID- 11474092 TI - Geology. The smile of the Cheshire cat. PMID- 11474091 TI - Immunology. The push-me pull-you of T cell activation. PMID- 11474093 TI - Filling generation gaps. PMID- 11474094 TI - An experiment for all seasons. PMID- 11474095 TI - Where the grass never stops growing. PMID- 11474096 TI - Divining a forest's future from its past. PMID- 11474097 TI - The partitioning of the Red Sea. PMID- 11474098 TI - Historical overfishing and the recent collapse of coastal ecosystems. AB - Ecological extinction caused by overfishing precedes all other pervasive human disturbance to coastal ecosystems, including pollution, degradation of water quality, and anthropogenic climate change. Historical abundances of large consumer species were fantastically large in comparison with recent observations. Paleoecological, archaeological, and historical data show that time lags of decades to centuries occurred between the onset of overfishing and consequent changes in ecological communities, because unfished species of similar trophic level assumed the ecological roles of overfished species until they too were overfished or died of epidemic diseases related to overcrowding. Retrospective data not only help to clarify underlying causes and rates of ecological change, but they also demonstrate achievable goals for restoration and management of coastal ecosystems that could not even be contemplated based on the limited perspective of recent observations alone. PMID- 11474099 TI - Noisy clockwork: time series analysis of population fluctuations in animals. AB - Both biotic interactions and abiotic random forcing are crucial influences on population dynamics. This frequently leads to roughly equal importance of deterministic and stochastic forces. The resulting tension between noise and determinism makes ecological dynamics unique, with conceptual and methodological challenges distinctive from those in other dynamical systems. The theory for stochastic, nonlinear ecological dynamics has been developed alongside methods to test models. A range of dynamical components has been considered-density dependence, environmental and demographic stochasticity, and climatic forcing-as well as their often complex interactions. We discuss recent advances in understanding ecological dynamics and testing theory using long-term data and review how dynamical forces interact to generate some central field and laboratory time series. PMID- 11474100 TI - Complex species interactions and the dynamics of ecological systems: long-term experiments. AB - Studies that combine experimental manipulations with long-term data collection reveal elaborate interactions among species that affect the structure and dynamics of ecosystems. Research programs in U.S. desert shrubland and pinyon juniper woodland have shown that (i) complex dynamics of species populations reflect interactions with other organisms and fluctuating climate; (ii) genotype x environment interactions affect responses of species to environmental change; (iii) herbivore-resistance traits of dominant plant species and impacts of "keystone" animal species cascade through the system to affect many organisms and ecosystem processes; and (iv) some environmental perturbations can cause wholesale reorganization of ecosystems because they exceed the ecological tolerances of dominant or keystone species, whereas other changes may be buffered because of the compensatory dynamics of complementary species. PMID- 11474101 TI - Long-term studies of vegetation dynamics. AB - By integrating a wide range of experimental, comparative, and theoretical approaches, ecologists are starting to gain a detailed understanding of the long term dynamics of vegetation. We explore how patterns of variation in demographic traits among species have provided insight into the processes that structure plant communities. We find a common set of mechanisms, derived from ecological and evolutionary principles, that underlie the main forces shaping systems as diverse as annual plant communities and tropical forests. Trait variation between species maintains diversity and has important implications for ecosystem processes. Hence, greater understanding of how Earth's vegetation functions will likely require integration of ecosystem science with ideas from plant evolutionary, population, and community ecology. PMID- 11474102 TI - Investigating long-term ecological variability using the Global Population Dynamics Database. AB - The Global Population Dynamics Database (GPDD) is an important new source of information for ecologists, resource managers, and environmental scientists interested in the dynamics of natural populations. It comprises more than 4500 time series of population abundance for over 1800 animal species across many taxonomic groups and geographical locations. The GPDD offers great potential for asking comparative questions about the nature of population variability. We illustrate this by characterizing some critical features of ecological variability, variance growth, and spectral reddening. PMID- 11474103 TI - Ecological forecasts: an emerging imperative. AB - Planning and decision-making can be improved by access to reliable forecasts of ecosystem state, ecosystem services, and natural capital. Availability of new data sets, together with progress in computation and statistics, will increase our ability to forecast ecosystem change. An agenda that would lead toward a capacity to produce, evaluate, and communicate forecasts of critical ecosystem services requires a process that engages scientists and decision-makers. Interdisciplinary linkages are necessary because of the climate and societal controls on ecosystems, the feedbacks involving social change, and the decision making relevance of forecasts. PMID- 11474104 TI - The composite genome of the legume symbiont Sinorhizobium meliloti. AB - The scarcity of usable nitrogen frequently limits plant growth. A tight metabolic association with rhizobial bacteria allows legumes to obtain nitrogen compounds by bacterial reduction of dinitrogen (N2) to ammonium (NH4+). We present here the annotated DNA sequence of the alpha-proteobacterium Sinorhizobium meliloti, the symbiont of alfalfa. The tripartite 6.7-megabase (Mb) genome comprises a 3.65-Mb chromosome, and 1.35-Mb pSymA and 1.68-Mb pSymB megaplasmids. Genome sequence analysis indicates that all three elements contribute, in varying degrees, to symbiosis and reveals how this genome may have emerged during evolution. The genome sequence will be useful in understanding the dynamics of interkingdom associations and of life in soil environments. PMID- 11474105 TI - Optical response of high-dielectric-constant perovskite-related oxide. AB - Optical conductivity measurements on the perovskite-related oxide CaCu3Ti4O12 provide a hint of the physics underlying the observed giant dielectric effect in this material. A low-frequency vibration displays anomalous behavior, implying that there is a redistribution of charge within the unit cell at low temperature. At infrared frequencies (terahertz), the value for the dielectric constant is approximately 80 at room temperature, which is far smaller than the value of approximately 10(5) obtained at lower radio frequencies (kilohertz). This discrepancy implies the presence of a strong absorption at very low frequencies due to dipole relaxation. At room temperature, the characteristic relaxation times are fast (less than or approximately 500 nanoseconds) but increase dramatically at low temperature, suggesting that the large change in dielectric constant may be due to a relaxor-like dynamical slowing down of dipolar fluctuations in nanosize domains. PMID- 11474106 TI - beta -Helical polymers from isocyanopeptides. AB - Polymerization of isocyanopeptides results in the formation of high molecular mass polymers that fold in a proteinlike fashion to give helical strands in which the peptide chains are arranged in beta-sheets. The beta-helical polymers retain their structure in water and unfold in a cooperative process at elevated temperatures. The peptide architecture in these polymers is a different form of the beta-helix motif found in proteins. Unlike their natural counterparts, which contain arrays of large beta-sheets stacked in a helical fashion, the isocyanopeptide polymers have a central helical core that acts as a director for the beta-sheet-like arrangement of the peptide side arms. The helical structure of these isocyanopeptide polymers has the potential to be controlled through tailoring of the side branches and the hydrogen-bonding network present in the beta-sheets. PMID- 11474107 TI - Topochemical polymerization of C70 controlled by monomer crystal packing. AB - Polymeric forms of C60 are now well known, but numerous attempts to obtain C70 in a polymeric state have yielded only dimers. Polymeric C70 has now been synthesized by treatment of hexagonally packed C70 single crystals under moderate hydrostatic pressure (2 gigapascals) at elevated temperature (300 degrees C), which confirms predictions from our modeling of polymeric structures of C70. Single-crystal x-ray diffraction shows that the molecules are bridged into polymeric zigzag chains that extend along the c axis of the parent structure. Solid-state nuclear magnetic resonance and Raman data provide evidence for covalent chemical bonding between the C70 cages. PMID- 11474108 TI - Calibration of the lutetium-hafnium clock. AB - Well-defined constants of radioactive decay are the cornerstone of geochronology and the use of radiogenic isotopes to constrain the time scales and mechanisms of planetary differentiation. Four new determinations of the lutetium-176 decay constant (lambda176Lu) made by calibration against the uranium-lead decay schemes yield a mean value of 1.865 +/- 0.015 x 10(-11) year(-1), in agreement with the two most recent decay-counting experiments. Lutetium-hafnium ages that are based on the previously used lambda176Lu of 1.93 x 10(-11) to 1.94 x 10(-11) year(-1) are thus approximately 4% too young, and the initial hafnium isotope compositions of some of Earth's oldest minerals and rocks become less radiogenic relative to bulk undifferentiated Earth when calculated using the new decay constant. The existence of strongly unradiogenic hafnium in Early Archean and Hadean zircons implies that enriched crustal reservoirs existed on Earth by 4.3 billion years ago and persisted for 200 million years or more. Hence, current models of early terrestrial differentiation need revision. PMID- 11474109 TI - Very-long-period seismic signals and caldera formation at Miyake Island, Japan. AB - Over a period of roughly 40 days, starting on 8 July 2000, a caldera structure 1.7 kilometers in diameter developed by means of gradual depression and expansion of the summit crater at Miyake Island, Japan. At the same time, very-long-period (VLP) seismic signals were observed once or twice a day. Source mechanism analyses of the VLP signals show that the moment tensor solutions are smooth step functions over a time scale of 50 seconds, with dominant volumetric change components. We developed a model to explain the caldera and the VLP signals, in which a vertical piston of solid materials in the conduit is intermittently sucked into the magma chamber by lateral magma outflow. This model offers potential for making quantitative estimations of the characteristic physical properties of magma systems. PMID- 11474110 TI - Egalitarianism in female African lions. AB - Because most cooperative societies are despotic, it has been difficult to test models of egalitarianism. Female African lions demonstrate a unique form of plural breeding in which companions consistently produce similar numbers of surviving offspring. Consistent with theoretical predictions from models of reproductive skew, female lions are unable to control each other's reproduction because of high costs of fighting and low access to each other's newborn cubs. A female also lacks incentives to reduce her companions' reproduction, because her own survival and reproduction depend on group territoriality and synchronous breeding. Consequently, female relationships are highly symmetrical, and female lions are "free agents" who only contribute to communal care when they have cubs of their own. PMID- 11474111 TI - Modeling household transmission of American trypanosomiasis. AB - American trypanosomiasis, or Chagas disease, caused by the protozoan parasite Trypanosoma cruzi and transmitted by blood-feeding triatomine bugs, is a chronic, frequently fatal infection that is common in Latin America. Neither adequate drugs nor a vaccine is available. A mathematical model calibrated to detailed household data from three villages in northwest Argentina shows that householders could greatly reduce the risk of human infection by excluding domestic animals, especially infected dogs, from bedrooms; removing potential refuges for bugs from walls and ceilings; and using domestically applied insecticides. Low-cost, locally practicable environmental management combined with intermittent use of insecticides can sustainably control transmission of T. cruzi to humans in rural Argentina and probably elsewhere. PMID- 11474112 TI - An autoinhibitory mechanism for nonsyntaxin SNARE proteins revealed by the structure of Ykt6p. AB - Ykt6p is a nonsyntaxin SNARE implicated in multiple intracellular membrane trafficking steps. Here we present the structure of the NH2-terminal domain of Ykt6p (Ykt6pN, residues 1 to 140). The structure of Ykt6pN differed entirely from that of syntaxin and resembled the overall fold of the actin regulatory protein, profilin. Like some syntaxins, Ykt6p adopted a folded back conformation in which Ykt6pN bound to its COOH-terminal core domain. The NH2-terminal domain plays an important biological role in the function of Ykt6p, which in vitro studies revealed to include influencing the kinetics and proper assembly of SNARE complexes. PMID- 11474113 TI - Lysophosphatidylcholine as a ligand for the immunoregulatory receptor G2A. AB - Although the biological actions of the cell membrane and serum lipid lysophosphatidylcholine (LPC) in atherosclerosis and systemic autoimmune disease are well recognized, LPC has not been linked to a specific cell-surface receptor. We show that LPC is a high-affinity ligand for G2A, a lymphocyte-expressed G protein-coupled receptor whose genetic ablation results in the development of autoimmunity. Activation of G2A by LPC increased intracellular calcium concentration, induced receptor internalization, activated ERK mitogen-activated protein kinase, and modified migratory responses of Jurkat T lymphocytes. This finding implicates a role for LPC-G2A interaction in the etiology of inflammatory autoimmune disease and atherosclerosis. PMID- 11474114 TI - Role of inorganic polyphosphate in promoting ribosomal protein degradation by the Lon protease in E. coli. AB - Inorganic polyphosphate (polyP), a polymer of hundreds of phosphate (Pi) residues, accumulates in Escherichia coli in response to stresses, including amino acid starvation. Here we show that the adenosine 5'-triphosphate-dependent protease Lon formed a complex with polyP and degraded most of the ribosomal proteins, including S2, L9, and L13. Purified S2 also bound to polyP and formed a complex with Lon in the presence of polyP. Thus, polyP may promote ribosomal protein degradation by the Lon protease, thereby supplying the amino acids needed to respond to starvation. PMID- 11474115 TI - The crystal structure of uncomplexed actin in the ADP state. AB - The dynamics and polarity of actin filaments are controlled by a conformational change coupled to the hydrolysis of adenosine 5'-triphosphate (ATP) by a mechanism that remains to be elucidated. Actin modified to block polymerization was crystallized in the adenosine 5'-diphosphate (ADP) state, and the structure was solved to 1.54 angstrom resolution. Compared with previous ATP-actin structures from complexes with deoxyribonuclease I, profilin, and gelsolin, monomeric ADP-actin is characterized by a marked conformational change in subdomain 2. The successful crystallization of monomeric actin opens the way to future structure determinations of actin complexes with actin-binding proteins such as myosin. PMID- 11474116 TI - Islet amyloid development in a mouse strain lacking endogenous islet amyloid polypeptide (IAPP) but expressing human IAPP. AB - BACKGROUND: Several mouse strains expressing human islet amyloid polypeptide (IAPP) have been created to study development of islet amyloid and its impact on islet cell function. The tendency to form islet amyloid has varied strongly among these strains by factors that have not been elucidated. Because some beta cell granule components are known to inhibit IAPP fibril formation in vitro, we wanted to determine whether a mouse strain expressing human IAPP but lacking the nonamyloidogenic mouse IAPP is more prone to develop islet amyloidosis. MATERIALS AND METHODS: Such a strain was created by cross-breeding a transgenic mouse strain and an IAPP null mouse strain. RESULTS: When fed a fat-enriched diet, male mice expressing only human IAPP developed islet amyloid earlier and to a higher extent than did mice expressing both human and mouse IAPP. Supporting these results, we found that mouse IAPP dose-dependently inhibits formation of fibrils from human IAPP. CONCLUSIONS: Female mice did not develop amyloid deposits, although small extracellular amorphous IAPP deposits were found in some islets. When cultivated in vitro, amyloid deposits occurred within 10 days in islets from either male or female mice expressing only human IAPP. The study shows that formation of islet amyloid may be dependent on the environment, including the presence or absence of fibril inhibitors or promoters. PMID- 11474117 TI - Potent induction of apoptosis by beta-lapachone in human multiple myeloma cell lines and patient cells. AB - BACKGROUND: Human multiple myeloma (MM) remains an incurable hematological malignancy. We have reported that beta-lapachone, a pure compound derived from a plant, can induce cell death in a variety of human carcinoma cells, including ovary, colon, lung, prostate, pancreas, and breast, suggesting a wide spectrum of anticancer activity. MATERIALS AND METHODS: We first studied antisurvival effects of beta-lapachone in human MM cells by colony formation assay. To determine whether the differential inhibition of colony formation occurs through antiproliferative activity, we performed MTT assays. The cytotoxicity of beta lapachone on human peripheral blood mononuclear cells was also measured by MTT assay. To determine whether the cell death induced by beta-lapachone occurs through necrosis or apoptosis, we used the propidium iodide staining procedure to determine the sub-GI fraction, Annexin-V staining for externalization of phosphatidylserine, and fragmentation of cellular genomic DNA subjected to gel electrophoresis. To investigate the mechanism of anti-MM activity, we examined Bcl-2 expression, cytochrome C release, and poly (ADP ribose) polymerase cleavage by Western blot assay. RESULTS: We found that beta-lapachone (less than 4 microM) inhibits cell survival and proliferation by triggering cell death with characteristics of apoptosis in ARH-77, HS Sultan, and MM.1S cell lines, in freshly derived patient MM cells (MM.As), MM cell lines resistant to dexamethasone (MM.1R), doxorubicin (DOX.40), mitoxantrone (MR.20), and mephalan (LR5). Importantly, after treatment with beta-lapachone, we observed no apoptosis in peripheral blood mononuclear cells in either quiescent or proliferative states, freshly isolated from healthy donors. In beta-lapachone treated ARH-77, cytochrome C was released from mitochondria to cytosol, and poly (ADP ribose) polymerase was cleaved, signature events of apoptosis. Finally, the apoptosis induced by beta-lapachone in MM cells was not blocked by either interleukin-6 or Bcl-2, which confer multidrug resistance in MM. CONCLUSIONS: Our results suggest potential therapeutic application of beta-lapachone against MM, particularly to overcome drug resistance in relapsed patients. PMID- 11474118 TI - Lack of interferon-gamma production despite the presence of interleukin-18 during cutaneous wound healing. AB - BACKGROUND: Recently, we have reported a rapid and strong induction of interleukin-18 (IL-18) upon cutaneous injury in mice. In this paper, we investigated a possible role of IL-18 in triggering interferon-gamma (IFN-gamma) production at the wound site. MATERIALS AND METHODS: Expression of IFN-gamma during cutaneous wound healing was analyzed by RNase protection assay, Western blot, ELISA, and immunohistochemical techniques in a murine model of excisional skin repair. RESULTS: We could not detect any IFN-gamma mRNA and protein expression during normal skin repair. Additionally, impaired healing in the genetically diabetic db/db mouse, which was used as a model for a prolonged inflammatory phase of repair, was characterized by largely elevated levels of IL 18 during the late phase of repair and an absence of IFN-gamma. Western blot analysis for T-cell- and monocyte/macrophage-specific marker proteins (CD4, F4/80) clearly revealed the presence of these subsets of leukocytic cells at the wound site, that are known to produce IFN-gamma in response to IL-18. Furthermore, we provide evidence that the presence of transforming growth factor beta1 (TGF-beta1) at the wound site might reflect a counterregulatory mechanism in IL-18-induced IFN-gamma production, as TGF-beta1 strongly suppressed IL 18/phytohaemagglutinin (PHA)-induced IFN-gamma production by peripheral blood mononuclear cells (PBMC) in vitro. CONCLUSIONS: Normal tissue regeneration processes after cutaneous injury were not dependent on the presence of IFN-gamma in vivo, and IL-18 must serve additional roles rather than inducing IFN-gamma during the healing process. PMID- 11474119 TI - Tumor necrosis factor and reactive oxygen species cooperative cytotoxicity is mediated via inhibition of NF-kappaB. AB - BACKGROUND: Tumor necrosis factor alpha (TNFalpha) plays a key role in pathogenesis of brain injury. However, TNFalpha exhibits no cytotoxicity in primary cultures of brain cells. This discrepancy suggests that other pathogenic stimuli that exist in the setting of brain injury precipitate TNFalpha cytotoxicity. The hypothesis was tested that reactive oxygen species (ROS), that are released early after brain injury, act synergistically with TNFalpha in causing cell death. MATERIALS AND METHODS: Cultured human and rat brain capillary endothelial cells (RBEC), and cortical astrocytes were treated with TNFalpha alone or together with different doses of H2O2, and apoptotic cell death and DNA fragmentation were measured by means of 3'-OH-terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and Hoechst fluorescence assay, respectively. The effect of H2O2 on TNFalpha-induced activation of nuclear factor kappa B (NF-kappaB) was measured by Western blots of cytoplasmic and nuclear extracts of RBEC using anti-inhibitor of NF-kappaB (IkappaB) and anti-p65 subunit of NF-kappaB antibodies. Nuclear translocation of NF-kappaB was investigated by immunofluorescent staining of RBEC with anti-p65 antibodies. RESULTS: TNFalpha alone had no cytotoxic effect in brain endothelial cells and astrocytes at concentrations up to 100 ng/ml. Co-treatment with 5-10 microM of H2O2 caused a two-fold increase in the number of apoptotic cells 24 hr later. Similar doses (1-3 microM) of H2O2 initiated early DNA fragmentation. H2O2 inhibited TNFalpha-induced accumulation of p65 in the nucleus, although it had no effect on degradation of the IkappaB in cytoplasm. Immunostaining confirmed that H2O2 inhibited p65 transport to the nucleus. CONCLUSIONS: Reactive oxygen species could act synergistically with TNFalpha in causing cytotoxicity via inhibition of a cytoprotective branch of TNFalpha signaling pathways, which starts with NF kappaB activation. PMID- 11474120 TI - Fc-receptor-mediated intracellular delivery of Cu/Zn-superoxide dismutase (SOD1) protects against redox-induced apoptosis through a nitric oxide dependent mechanism. AB - BACKGROUND: Using specific antibodies against bovine Cu/Zn-superoxide dismutase (EC 1.15.1.1, SOD1) we demonstrated that anti-SOD antibodies (IgG1) are able to promote the intracellular translocation of the antioxidant enzyme. The transduction signalling mediated by IgG1 immune complexes are known to promote a concomitant production of superoxide and nitric oxide leading to the production of peroxynitrites and cell death by apoptosis. The Fc-mediated intracellular delivery of SOD1 thus limited the endogenous production of superoxide. It was thus of interest to confirm that in the absence of superoxide anion, the production of nitric oxide protected cells against apoptosis. Study in greater detail clearly stated that under superoxide anion-free conditions, nitric oxide promoted the cell antioxidant armature and thus protected cells against redox induced apoptosis. MATERIALS AND METHODS: The murine macrophage cell-lines J774 A1 were preactivated or not with interferon-gamma and were then stimulated by IgG1 immune complexes (IC), free SOD1 or SOD1 IC and superoxide anion, nitric oxide, peroxynitrite, and tumor necrosis factor-alpha (TNF-alpha) production was evaluated. The redox consequences of these activation processes were also evaluated on mitochondrial respiration and apoptosis as well as on the controlled expression of the cellular antioxidant armature. RESULTS: We demonstrated that SOD1 IC induced a Fcgamma receptor (FcgammaR)-dependent intracellular delivery of the antioxidant enzyme in IFN-gamma activated murine macrophages (the J774 AI cell line). The concomitant stimulation of the FcyR and the translocation of the SOD1 in the cytoplasm of IFN-gamma-activated macrophages not only reduced the production of superoxide anion but also induced the expression of the inducible form of nitric oxide synthase (iNOS) and the related NO production. This inducing effect in the absence of superoxide anion production reduced mitochondrial damages and cell death by apoptosis and promoted the intracellular antioxidant armature. CONCLUSIONS: To define the pharmacologic mechanism of action of bovine SOD1, we attempted to identify the second messengers that are induced by SOD1 IC. In this work, we propose that Fc-mediated intracellular delivery of the SOD1 that reduced the production of superoxide anion and of peroxynitrite, promoted a NO induced protective effect in inducing the antioxidant armature of the cells. Taken together, these data suggested that specific immune responses against antigenic SOD1 could promote the pharmacological properties of the antioxidant enzyme likely via a NO-dependent mechanism. PMID- 11474121 TI - New insight into mechanisms of allograft transplantation in the rat by differential display: macrophage scavenger receptor-A brings to light. AB - BACKGROUND: Donor specific tolerance to heart allografts is induced in LEW.1A rat recipient by two donor LEW.1W blood transfusions prior engraftment. Although the tolerant allograft is infiltrated by leukocytes, graft infiltrating cells are only expressing low levels of the Th1- or Th2-related cytokines suggesting that induction of tolerance is an active phenomenon in which the mechanisms remain to be elucidated. MATERIALS AND METHODS: Differential display (DD) method was applied on RNAs extracted from graft infiltrating cells (GIC) derived from allografts either from rejecting untreated rats or donor-specific blood transfusion treated tolerant rats. Quantitative RT/PCR was performed to confirm mRNA expressions of the selected genes. RESULTS: Among the six differentially displayed DNAs (ddDNA) overexpressed in GIC from rejected allografts, the macrophage scavenger receptor-A (A:D13265) was identified; it exhibited a stricking induction of mRNA expression from day 1 to 7 after transplantation. Among the seven ddDNAs overexpressed in GIC from tolerant allografts, the 3 hydroxy-3-methyl glutaryl coenzyme-A reductase (A:M29249) and an "unknown gene" (ddDNA EC9) were identified and both were confirmed to be up-regulated by quantitative RT/PCR. CONCLUSIONS: The relevance of these genes in transplantation has not yet been reported and must therefore be elucidated; they represent possible targets for immunointervention. Nevertheless, our data demonstrate that the DD is a powerful tool to identify new genes involved in organ transplantation. PMID- 11474122 TI - Essential components of antimicrobial gastrointestinal epithelial barrier: specific interaction of mucin with an integral apical membrane protein of gastric mucosa. AB - BACKGROUND: The gastric mucosal protective barrier consists of two essential elements: mucus glycoprotein, mucin, secreted by gastric mucosal cells, and the mucin binding protein (MBP), an integral component of the apical epithelial membrane. The studies described here provide evidence on the structure of MBP, its interaction with mucin, and the susceptibility to phospholipase C (PLC) and Helicobacter pylori protease. MATERIAL AND METHODS: The rat gastric mucosa was used to isolate mucin and the apical epithelial membranes. A buffered saline extract of the mucosal cells was used for the isolation of mucin and the 1% Triton X-100-insoluble gastric apical membranes for the preparation of MBP. RESULTS: The studies on MBP, the mucosal mucin receptor revealed that the protein is anchored in apical membrane through glycosylphosphatidylinositol (GPI). The deamination of MBP with nitrous acid afforded phosphatidylinositols (PIs) and a water soluble, 97 kDa glycosylated protein. The in situ studies with untreated rat gastric mucosa and the mucosa depleted of mucin showed that MBP without mucin was susceptible to the proteolytic degradation with pepsin and H. pylori proteases, but was not released from the apical membrane by the treatment with bacterial PLC. CONCLUSION: The study of carbohydrate ligands for MBP revealed binding of octa- and decasaccharides of gastric mucin. The severe impairment in mucin adhesion to MBP, induced by the diet containing ethanol, supports the conclusion that specific carbohydrate determinants participate in mucin attachment to MBP and epigenetic control of the processes that coordinates its interaction with apical mucosal epithelium in the formation of innate protective barrier. PMID- 11474123 TI - Vdelta1 T lymphocytes expressing a Th1 phenotype are the major gammadelta T cell subset infiltrating the liver of HCV-infected persons. AB - BACKGROUND: Hepatitis C infection induces an acute and chronic liver inflammation that may lead to cirrhosis, liver failure, or hepatocarcinoma. Since the role of alphabeta T lymphocytes in hepatitis C virus (HCV) immunopathology has been analyzed extensively, we investigated the distribution and functional activation of gammadelta T cell subsets in chronically HCV-infected patients. MATERIALS AND METHODS: Blood samples and liver biopsies from 35 patients with compensated chronic HCV infection were compared in terms of T cell subset distribution, expression of activation markers, gammadelta T cell receptor (TCR) repertoire, and pattern of cytokine production. Moreover, we analyzed whether these immunological parameters were associated with other clinical observations (plasma viremia, ALT levels, Ishak index). RESULTS: Differing from peripheral blood distribution, a specific compartmentalization of Vdelta1 T cells (p < 0.001) was observed in the liver of HCV patients. These cells represented a relevant fraction of intrahepatic T lymphocytes (1.8-8.7%) and expressed the memory/effector phenotype (CD62-L- CD45-RO+CD95+). This phenotype was consistent with selective homing upon antigen recognition. Mitogenic stimulation of Vdelta1 + T lymphocytes recruited in the liver revealed the T helper cell type 1 (Th1) pattern of cytokine secretion. Interestingly, the frequency of interferon-gamma (IFN-gamma)-producing Vdelta1 T cells was associated with an higher degree of liver necroinflammation, measured by the Ishak index. Finally, the T-cell repertoire analysis revealed the absence of Vgamma selection in the TCR repertoire of intrahepatic Vdelta1 T cells. CONCLUSIONS: gammadelta T cell distribution in the peripheral blood differs from the Vdelta1 T cell subset because it is policlonally activated and recruited in the liver of chronic HCV infected patients. During HCV-infection, this T cell subset may release Th1 cytokines and contribute to the necroinflammatory liver disease. PMID- 11474124 TI - Increased insulin-like growth factor 1 activity can rescue KLE endometrial-like cells from apoptosis. AB - BACKGROUND: The peritoneal fluid (PF) of women with endometriosis contains protease(s) activity able to hydrolyze insulin-like growth factor-binding protein 3 (IGFBP-3), increasing the bioavailability of insulin-like growth factor-1 (IGF 1) locally. Therefore, we characterized the effects of IGF-1 on KLE endometrial like cells in vitro. MATERIALS AND METHODS: The mitogenic effect of IGF-1 was assessed by the analysis of the DNA content and cell count. Apoptosis was triggered experimentally by the 48 hr exposure of KLE cells to 100 nM of adriamycin in the presence and absence of IGF-1 (50 ng/ml). Adriamycin apoptosis of KLE cells was determined by the number of dead KLE cells using trypan blue exclusion and by the DNA fragmentation on simple agarose gel and flow cytometry of propidium iodide and HOECHST 33342-stained KLE cells using an EPICS 753 pulse cytometer. RESULTS: IGF-1 stimulated the growth of KLE cells in a dose-dependent manner (optimal dose of 50 ng/ml) and protected KLE cells from adriamycin (100 nM)-induced apoptosis. CONCLUSIONS: These data suggest that IGF-1 is a survival factor for KLE cells. Conceivably, increased IGF-1 activity in the PF can optimize both the survival and ectopic growth of endometrial cells in the peritoneal cavity. PMID- 11474125 TI - Purification and characterization of macrophage migration inhibitory factor as a secretory protein from rat epididymis: evidences for alternative release and transfer to spermatozoa. AB - BACKGROUND: The cytokine macrophage migration inhibitory factor (MIF), originally described as a T cell product, has recently been identified to mediate cellular interactions in several endocrine organs. Western blots analysis of rat epididymal homogenates using an anti-MIF antibody indicated the presence of substantial amounts of an immunoreactive protein with the apparent Mr of 12 kDa. Our study aimed to characterize the molecular nature of this immunoreactive factor. MATERIALS AND METHODS: The purified 12 kDa protein and a cloned cDNA fragment were characterized by sequence analysis. Furthermore, expression pattern and localization of the 12 kDa protein were investigated using in situ hybridization, immunohistochemistry, immunoelectron microscopy, and western blots experiments on epididymal sections, isolated epididymal vesicles, and outer dense fibers from spermatozoa. RESULTS: The N-terminal amino acid sequence analysis over 10 amino acids revealed a 100% homology of the 12 kDa protein to the N terminus of the cytokine MIF. These data were confirmed by sequence analysis of a reverse transcription polymerase chain reaction (RT-PCR) amplified cDNA fragment from rat epididymis, which also showed complete homology to the MIF cDNA sequence. MIF protein and mRNA were localized in the epithelial cells of the epididymis in a regional distribution manner, with the expression maximal in the caput. Immune cells were not labeled. MIF is the first classical cytokine identified to be expressed by the epididymal epithelial cells. Immunoelectron microscopy detected MIF immunoreactivity in the cytoplasm, with no reaction visible in the Golgi complex and the cisternae of the endoplasmic reticulum. At the apical cell surface, MIF accumulated in stereocilia and vesicles that were pinched off from the plasma membrane. MIF detection in vesicles isolated from epididymal secretion together with the lack of a N-terminal signal sequence for translocation in the endoplasmic reticulum strongly suggested a nonclassical secretion mode. Furthermore, MIF was identified as a new component of the outer dense fibers (ODF), a cytoskeletal element of the mid- and principal piece of the sperm tail. CONCLUSION: The cytokine MIF was identified in substantial amounts in the epithelial cells of rat epididymis and in the outer dense fibers of rat epididymal spermatozoa. Our results indicate a nonclassical secretion mode for MIF and suggest a cell-to-cell transfer of MIF via vesicles to the sperm cells. PMID- 11474126 TI - Apoptosis in caspase-inhibited neurons. AB - BACKGROUND: There is growing evidence of apoptosis in neurodegenerative disease. However, it is still unclear whether the pathological manifestations observed in slow neurodegenerative diseases are due to neuronal loss or whether they are related to independent degenerative events in the axodendritic network. It also remains elusive whether a single, caspase-based executing system involving caspases is responsible for neuronal loss by apoptosis. MATERIALS AND METHODS: Long-term exposure to the microtubule-disassembling agent, colchicine, was used to disrupt the axodendritic network and eventually trigger caspase-3-mediated apoptosis in cultures of cerebellar granule cells. For this model, we investigated the role of Bcl-2 and caspases in neurite degeneration and death of neuronal somata. RESULTS: Early degeneration of the axodendritic network occurred by a Bcl-2 and caspase-independent mechanism. Conversely, apoptosis of the cell body was delayed by Bcl-2 and initially blocked by caspase inhibition. However, when caspase activity was entirely blocked by zVAD-fmk, colchicine-exposed neurons still underwent delayed cell death characterized by cytochrome c release, chromatin condensation to irregularly shaped clumps, DNA-fragmentation, and exposure of phosphatidylserine. Inhibitors of the proteasome reduced these caspase-independent apoptotic-like features of the neuronal soma. CONCLUSION: Our data suggest that Bcl-2-dependent and caspase-mediated death programs account only partially for neurodegenerative changes in injured neurons. Blockage of the caspase execution machinery may only temporarily rescue damaged neurons and classical apoptotic features can still appear in caspase-inhibited neurons. PMID- 11474127 TI - p27Kip1 localizes to detergent-insoluble microdomains within lymphocyte membranes. AB - BACKGROUND: Low levels of the cyclin-dependent kinase inhibitor p27Kip1 are associated with poor prognosis in cancer. It is unclear whether this is related strictly to p27Kip1-mediated cell cycle inhibition or to other, possibly extranuclear, roles of this protein. In this study, we examined p27Kip1 expression in quiescent and activated lymphocytes. T-cell membranes have been shown to possess sphingolipid and cholesterol-rich microdomains that are insoluble in non-ionic detergents. These "rafts" provide a scaffold for signaling proteins. Signal transduction coincides with coalescence of these microdomains into larger complexes. METHODS: Localization of p27Kip1 was studied by electron and confocal microscopy. Association of p27Kip1 with membrane microdomains in unstimulated and stimulated lymphocytes was determined using Western blots analysis of isolated membranes variably treated with detergents. RESULTS: We demonstrated that p27Kip1 was present in clusters associated with the plasma membrane in normal lymphocytes. The solubility profile of p27Kip1 in isolated membranes indicated that it was localized to raft structures. When lymphocytes were stimulated, however, p27Kip1 was excluded from aggregated raft complexes. CONCLUSIONS: This study identifies, for the first time, the localization of p27 within a membrane microdomain associated with signaling. Because some cell surface signaling complexes lose p27Kip1 upon cellular activation, p27Kip1 may play a functional role in modulating membrane signaling. PMID- 11474128 TI - A therapeutic target for hormone-independent estrogen receptor-positive breast cancers. AB - BACKGROUND: The action of the steroid hormone estradiol (E2) is mediated via interaction with a specific receptor (ER) that initiates a series of events downstream, leading to the modulation of hormone-responsive genes and cell proliferation. Antihormones also bind, but do not confer the active configuration to ER, thereby, blocking the transmission of E2-ER-initiated signals for cell proliferation. Although these compounds qualify for successful therapy of ER positive [ER (+)] breast cancer patients, only a fraction of patients responds to antihormone treatment. In this study, the functional status of ER is determined to identify alternative targets for therapy of antihormone-resistant ER (+) breast cancers. METHOD: The interaction of ER with a specific DNA sequence, designated as E2 response element (ERE), was targeted to assess the functional state of ER. ER-ERE complex formation was measured by electrophoretic mobility shift assay (EMSA) and by a newly developed technique, based on the preferential binding of DNA-protein complex to a nitrocellulose membrane (NMBA) that measures both total and functional fraction of ER. RESULTS: The NMBA assay identified functional variants of ER among ER (+) breast cancer cell lines and breast tumor biopsy specimens. ER of (21PT) cells did not bind E2 and these cells were tamoxifen (TAM) resistant. However 21PT cells were sensitive to a calmodulin (CaM) antagonist, W7, that blocked ERE-ER complex formation. CONCLUSIONS: ER variants of the 21PT type were detected among breast cancer biopsy specimens, emphasizing the significance of an alternative therapeutic target for TAM resistant ER (+) human breast cancers with compounds such as W7. PMID- 11474129 TI - Stretch-mediated activation of selective MAPK subtypes and potentiation of AP-1 binding in human osteoblastic cells. AB - BACKGROUND: It is widely accepted that mechanical loading is necessary to construct the architecture of bone and to maintain bone mass. However, the molecular mechanisms whereby bone cells respond to mechanical stimuli remain elusive. The mitogen-activated protein kinase (MAPK) signaling cascades are known to play a crucial role in the immediate osteoblast response to a variety of bone active agents. An important component of this response occurs at the transcriptional level and is executed by downstream phosphorylation substrates, most notably, a number of signal-responsive transcription factors. To identify whether the MAPKs are involved in the mechanotransduction process and to determine the effect on down-stream transcription factors, we stimulated human periodontal ligament (hPDL) osteoblast-like cells by mechanical stretching by employing an established in vitro model of continuous stretch application. MATERIALS AND METHODS: Whole-cell extracts were prepared from cultivated hPDL cells that were exposed to short-term, continuous mechanical stretch. In-gel kinase assays were used to assess their kinase activity towards the immediate early gene products c-Jun and c-Fos [constituents of the activator protein-1 (AP 1) transcription factor]. Electrophoretic mobility-shift and southwestern experiments utilizing a DNA sequence that contained a previously undefined atypical AP-1-binding site in the promoter of the human liver/bone/kidney alkaline phosphatase (L/B/K ALP) gene (an early marker for osteoblastic differentiation) were employed to evaluate their specific binding capacity. RESULTS: Selective members of the MAPK family were rapidly induced by stretching, as manifested by their ability to enhance phosphorylation of their cognate substrates c-Jun and, to a lesser extent, c-Fos in the in-gel kinase assay. This induction was accompanied by markedly increased, phospho-c-Jun-containing AP-1 binding activity, as determined by the binding analyses performed with the relevant sequence from the L/B/K ALP promoter. CONCLUSIONS: In as much as AP-1 is instrumental in regulating genes activated at the onset of osteoblast differentiation, such as the ALP gene, we pose that an interplay of distinct MAPKs targeting AP-1 components may dictate the osteogenic response of hPDL cells to mechanical stimulation. PMID- 11474130 TI - The role of the complement cascade in ischemia/reperfusion injury: implications for neuroprotection. AB - BACKGROUND: The complement cascade plays a deleterious role in multiple models of ischemia/reperfusion (I/R) injury, including stroke. Investigation of the complement cascade may provide a critical approach to identifying neuroprotective strategies that can be effective at clinically relevant time points in cerebral ischemia. This review of the literature describes the deleterious effects of complement activation in systemic I/R models and previous attempts at therapeutic complement inhibition, with a focus on the potential role of complement inhibition in ischemic neuroprotection. Translation of these concepts into ischemic stroke models and exploration of related neuroprotective strategies are also reviewed. SUMMARY OF REVIEW: We performed a MEDLINE search to identify any studies published between 1966 and 2001 dealing with complement activation in the setting of I/R injury. We also searched for studies demonstrating up-regulation of any complement components within the central nervous system during inflammation and/or ischemia. CONCLUSIONS: The temporal and mechanistic overlap of the complement cascade with other biochemical events occurring in cerebral I/R injury is quite complex and is only beginning to be understood. However, there is compelling evidence that complement is quite active in the setting of acute stroke, suggesting that anticomplement strategies should be further investigated through genetic analysis, nonhuman primate models, and clinical investigations. PMID- 11474131 TI - Gene expression in atherosclerotic lesion of ApoE deficient mice. AB - BACKGROUND: Atherosclerosis, the major cause of mortality and invalidity in industrialized countries, is a multifactorial disease associated with high plasma cholesterol levels and inflammation in the vessel wall. Many different genes have previously been demonstrated in atherosclerosis, although limited numbers of genes are dealt with in each study. In general, data on dynamic gene expression during disease progress is limited and large-scale evaluation of gene expression patterns during atherogenesis could lead to a better understanding of the key events in the pathogenesis of atherosclerosis. We have therefore applied a mouse gene filter array to analyze gene expression in atherosclerotic ApoE-deficient mice. MATERIALS AND METHODS: ApoE-deficient mice were fed atherogenic western diet for 10 or 20 weeks and aortas isolated. C57BL/6 mice on normal chow were used as controls. The mRNAs of 15 animals were pooled and hybridized onto commercially available Clontech mouse gene array filters. RESULTS: The overall gene expression in the ApoE-deficient and control mice correlated well at both time points. Gene expression profiling showed varying patterns including genes up regulated at 10 or 20 weeks only. At 20 weeks of diet, an increasing number of up regulated genes were found in ApoE-deficient mice. CONCLUSIONS: The gene expression in atherogenesis is not a linear process with a maximal expression at advanced lesion stage. Instead, several genes demonstrate a dynamic expression pattern with peaks at the intermediate lesions stage. Thus, detailed evaluation of gene expression at several time points should help understanding the development of atherosclerosis and establishment of preventive intervention. PMID- 11474132 TI - Factors secreted by human neuroblastoma mediated doxorubicin resistance by activating STAT3 and inhibiting apoptosis. AB - BACKGROUND: The transcription factor Stat3 has been reported to play a key role in protecting cells against apoptosis by up-regulating expression of the anti apoptotic gene BCl-xL. This investigation analyzes the relationship between the development of resistance to doxorubicin-mediated apoptosis in neuroblastoma cells (SKN-SH) and activation of the Stat3 signaling pathway. MATERIALS AND METHODS: A drug-resistant cell line (SKN-SH/Dox6) was generated by continuous exposure to incremental concentrations of doxorubicin. Specific antibodies were utilized for Western blots and confocal microscopy to determine the nuclear localization of activated Stat3. RESULTS: Doxorubicin-mediated DNA fragmentation was inhibited and caspase-3 activity decreased in SKN-SH/Dox6 cells. Up regulation of Stat3 phosphorylation and Bcl-xL expression, increased nuclear translocation of phospho-Stat3, and binding to DNA occurred only in resistant SKN SH/Dox6 cells. The expression of Bcl-xL was inhibited by AG490, an inhibitor of the JAK/Stat3 signaling pathway, suggesting that the regulation of Bcl-xL and Stat3 involved a common mechanism. Activation of Stat3 in SKN-SH/Dox6 cells was contingent upon stimulation evoked by ligands secreted by the drug-resistant cells. Evidence to support this hypothesis was provided by experiments in which doxorubicin-sensitive SKN-SH cells were preincubated with conditioned media obtained from doxorubicin-resistant SKN-SH/Dox6 cells. This treatment increased Stat3 activation. It also rendered SKN-SH cells resistant to doxorubicin as demonstrated by a sharp decrease in doxorubicin-induced DNA degradation and cytotoxic potency. CONCLUSIONS: These findings suggest that the resistance of SKN SH/Dox6 cells to doxorubicin may be mediated by anti-apoptotic factor(s) that are synthesized and secreted by tumor cells in response to cytotoxic agents. PMID- 11474133 TI - Acetaldehyde adducts in blood and bone marrow of patients with ethanol-induced erythrocyte abnormalities. AB - BACKGROUND: Although alcohol abuse is known to cause a wide array of adverse effects on blood cell formation, the molecular mechanisms by which alcohol exerts its toxic actions remain poorly defined. We examine here the formation of acetaldehyde-derived protein modifications in erythrocytes and in their bone marrow precursors using antibodies specifically recognizing acetaldehyde-modified epitopes in proteins independently of the nature of the carrier protein. MATERIALS AND METHODS: We studied 138 consecutive adult patients undergoing bone marrow aspiration due to macrocytosis (MCV values above 99 fL). Assessment included complete blood counts, morphologic review, assessment of alcohol consumption, and biochemical and immunocytochemical assays for acetaldehyde adducts. RESULTS: There were 68 patients (49%) with a history of excessive alcohol consumption, 28 (20%) of whom were patients with severe dependence. The blood smears prepared from the alcoholic patients with macrocytosis also contained stomatocytes and knizocytes. Bone marrow aspirates from 12 alcoholic patients showed vacuolization of pronormoblasts and the presence of ring sideroblasts was noted in 8 cases. In immunocytochemical analyses of the peripheral blood erythrocytes, acetaldehyde-derived epitopes were found to occur both on the cell membrane and inside the erythrocytes. Bone marrow aspirates also showed positive staining for acetaldehyde adducts in the erythropoietic cells in 8 of 11 (73%) consecutive alcoholic patients. Separation of the erythrocyte proteins from the samples of alcoholics on HPLC-chromatography revealed the formation of fast-eluting hemoglobin fractions, which also reacted with antibodies against acetaldehyde adducts. CONCLUSIONS: Current data suggest that acetaldehyde-erythrocyte adducts are formed in vivo in blood and bone marrow of patients with excessive alcohol consumption. This may contribute to the generation of the erythrocyte abnormalities, which are frequently observed in alcoholic patients. PMID- 11474134 TI - Myocardial ischemic preconditioning in rodents is dependent on poly (ADP-ribose) synthetase. AB - BACKGROUND: Activation of the nuclear enzyme poly (ADP-ribose) synthetase (PARS) in response to oxidant-mediated DNA injury has been shown to play an important role in the pathogenesis of reperfusion injury. Here we investigated the role of PARS in myocardial ischemic preconditioning (IPC). MATERIALS AND METHODS: Mice with or without genetic disruption of PARS and rats in the absence or presence of the PARS inhibitor 3-aminobenzamide underwent coronary occlusion and reperfusion with or without IPC. RESULTS: Both poly(ADP-ribose) synthetase (PARS) deficiency and ischemic preconditioning (IPC) induced protection from reperfusion injury, attenuated inflammatory mediator production, and reduced neutrophil infiltration when compared to the response in wild-type mice. Surprisingly, the protective effect of IPC not only disappeared in PARS-/- mice, but the degree of myocardial injury and inflammatory response was similar to the one seen in wild-type animals. Similarly, in the rat model of IPC, 3-aminobenzamide pretreatment blocked the beneficial effect of IPC. Myocardial NAD+ levels were maintained in the PARS-deficient mice during reperfusion, while depleted in the wild-type mice. The protection against reperfusion injury by IPC was also associated with partially preserved myocardial NAD+ levels, indicating that PARS activation is attenuated by IPC. This conclusion was further strengthened by poly(ADP-ribose) immunohistochemical measurements, demonstrating that IPC markedly inhibits PARS activation during reperfusion. CONCLUSIONS: The mode of IPC's action is related, at least in part, to an inhibition of PARS. This process may occur either by self auto-ribosylation of PARS during IPC, and/or via the release of endogenous purines during IPC that inhibit PARS activation during reperfusion. PMID- 11474135 TI - Low levels of p27 in association with deregulated p53-pRb protein status enhance tumor proliferation and chromosomal instability in non-small cell lung carcinomas. AB - BACKGROUND: Down-regulation or overexpression of the cyclin-dependent kinase inhibitor p27 have been observed in a range of malignancies, including lung cancer. To further elucidate the role of the molecule in tumor growth regulation, we evaluated p27 expression in a series of non-small cell lung carcinomas (NSCLCs), and examined its relation with histology, kinetic parameters, ploidy, and overall survival. We extended our investigation into the association of p27 levels with the presence of Ki-ras mutations, as well as with the expression status of p53 and pRb in tumor cells. MATERIAL AND METHODS: p27, p53, and pRb status were immunohistochemically evaluated in a total of 69 NSCLCs. In situ assays were employed to assess the kinetic parameters (Ki-67 immunohistochemistry for proliferation index, Tdt-mediated dUTP nick end labeling assay for apoptotic index). The ploidy status of the tumors was assessed after staining nuclei with the Feulgen procedure, and the presence of Ki-ras mutations was examined by restriction fragment length polymorphisms. All possible associations were assessed with a series of statistical methods. RESULTS: Immunoreactivity for p27 was observed in the entire series of specimens, with the mean percentage of positive cells being 33%. Adenocarcinomas (AdCs) exhibited higher p27 levels compared to squamous cell carcinomas (SqCCs) (p < 0.01). An inverse correlation was established between p27 expression and proliferation index (PI) (r = -0.834, p < 0.01) but not with apoptotic index (AI), whereas aneuploid tumors were characterized by lower p27 levels than diploid ones (p < 0.01). No difference in p27 immunostaining was observed with regard to the presence of Ki-ras mutations, whereas aberrant p53 and/or pRb expression patterns were associated with p27 underexpression (p < 0.01 for p53 status, p < 0.05 regarding pRb levels, and p < 0.01 for a combined deregulation of both proteins). Two or more alterations in the p27/p53/pRb protein network (i.e., p27 levels lower than the estimated mean value, overexpressed p53, and/or aberrant pRb) were associated with increased PI and aneuploidy (p < 0.001 and p < 0.01, respectively). A powerful trend was found between p27 expression and overall survival (p = 0.066). CONCLUSIONS: Our findings confirm the heterogeneity between AdCs and SqCCs, and are suggestive of an increased proliferative activity in NSCLCs underexpressing p27. Furthermore, our analysis supports the concept of p27 forming a functionally compact network with p53 and pRb, which is actively involved in the regulation of cellular proliferation and chromosomal stability. PMID- 11474136 TI - The pathway enabling external sounds to reach and excite the fetal inner ear. AB - The human fetus in utero is able to respond to sounds in the amniotic fluid enveloping the fetus after about 20 weeks gestation. The pathway by which sound reaches and activates the fetal inner ear is not entirely known. It has been suggested that in this total fluid environment, the tympanic membrane and the round window membrane become 'transparent' to the sound field, enabling the sounds to reach the inner ear directly through the tympanic membrane and the round window membrane. It is also possible that sounds reach the inner ear by means of tympanic membrane--ossicular chain--stapes footplate conduction (as in normal air conduction). There is also evidence that sounds reach the fetal inner ear by bone conduction. Several animal and human models of the fetus in utero were studied here in order to investigate the pathway enabling sounds to reach and activate the fetal inner ear. This included studying the auditory responses to sound stimuli of animals and humans under water. It was clearly shown in all the models that the dominant mechanism was bone conduction, with little if any contribution from the external and middle ears. Based on earlier experiments on the mechanism and pathway of bone conduction, the results of this study lead to the suggestion that the skull bone vibrations induced by the sound field in the amniotic fluid enveloping the fetus probably give rise to a sound field within the fetal cranial cavity (brain and CSF) which reaches the fetal inner ear through fluid communication channels connecting the cranial cavity and the inner ear. PMID- 11474137 TI - Overexpression of copper/zinc-superoxide dismutase protects from kanamycin induced hearing loss. AB - The participation of reactive oxygen species in aminoglycoside-induced ototoxicity has been deduced from observations that aminoglycoside-iron complexes catalyze the formation of superoxide radicals in vitro and that antioxidants attenuate ototoxicity in vivo. We therefore hypothesized that overexpression of Cu/Zn-superoxide dismutase (h-SOD1) should protect transgenic mice from ototoxicity. Immunocytochemistry confirmed expression of h-SOD1 in inner ear tissues of transgenic C57BL/6-TgN[SOD1]3Cje mice. Transgenic and nontransgenic littermates received kanamycin (400 mg/kg body weight/day) for 10 days beginning on day 10 after birth. Auditory thresholds were tested by evoked auditory brain stem responses at 1 month after birth. In nontransgenic animals, the threshold in the kanamycin-treated group was 45-50 dB higher than in saline-injected controls. In the transgenic group, kanamycin increased the threshold by only 15 dB over the respective controls. The effects were similar at 12 and 24 kHz. The protection by overexpression of superoxide dismutase supports the hypothesis that oxidant stress plays a significant role in aminoglycoside-induced ototoxicity. The results also suggest transgenic animals as suitable models to investigate the underlying mechanisms and possible strategies for prevention. PMID- 11474138 TI - Functional beta2-adrenergic receptors are present in nonstrial tissues of the lateral wall in the gerbil cochlea. AB - Recently, we have demonstrated that functional beta1-adrenergic receptors are the dominant beta-adrenergic receptor subtype in the stria vascularis and that beta1 adrenergic receptors stimulate K+ secretion in strial marginal cells. The goal of the present study was to determine whether nonstrial tissues in the cochlear lateral wall contain beta-adrenergic receptors and if so which subtype is present. Pharmacological tools were used to identify receptors in functional studies where cAMP production was measured. Further, receptors were identified as transcripts by cloning and sequencing of reverse-transcriptase polymerase chain reaction (RT-PCR) products. Experiments were performed on gerbil nonstrial lateral wall tissues. Tissues stimulated with 10(-5) M isoproterenol produced 0.42 +/- 0.22 pmol cAMP per ear within 12 min (n = 14). The selective beta adrenergic receptor agonist isoproterenol stimulated cAMP production with an EC50 of (2 +/- 3) x 10(-7) M (n = 7). Isoproterenol-stimulated cAMP production was inhibited by the beta2-adrenergic receptor antagonist ICI 118551 with an IC50 of (7 +/- 7) x 10(-6) M, which corresponds to an affinity constant of 1 x 10(-7) M (pK(DB) = 6.89 +/- 0.23, n = 3). Isoproterenol-stimulated cAMP production was not inhibited by the highly selective beta1-adrenergic receptor antagonist CGP 20712A. The IC50 and the affinity constant for CGP 20712A were estimated to be >3 x 10(-4) and >6 x 10(-6) M, respectively. RT-PCR of total RNA isolated from nonstrial lateral wall tissues using primers specific for beta1-, beta2- and beta3-adrenergic receptors revealed products of the predicted sizes for the beta1 and beta2- but not for the beta3-subtype. Sequence analysis confirmed that amplified cDNA fragments encoded gene-specific nucleotide sequences. These results demonstrate that nonstrial lateral wall tissues contain transcripts for beta1- and beta2- but not for beta3-adrenergic receptors and that the beta2 adrenergic receptor is the dominant functional receptor subtype. The cellular localization and function of the beta2-adrenergic receptors remains to be determined. PMID- 11474139 TI - Peripheral cell loss related to calcium binding protein immunocytochemistry in the dorsal cochlear nucleus in CBA/CaJ mice during aging. AB - The influence of cochlear hair cell and spiral ganglia neuron loss on calcium binding protein immunoreactivity (calretinin, parvalbumin and calbindin) in the dorsal and posteroventral cochlear nuclei (DCN and PVCN) in CBA/CaJ (CBA) mice during aging (1-39 months) was determined. Since calcium binding proteins have buffering properties against calcium overload, they may have a protective role during aging. It is shown that the percentage of calretinin- and parvalbumin immunopositive neurons in the DCN showed a statistically significant positive correlation with inner hair cell loss, outer hair cell loss, and spiral ganglion cell loss. A correlation was also found between aging and the auditory periphery, and calcium binding proteins in the DCN. These findings imply that the pathophysiological state of the auditory periphery may influence the neuronal homeostasis in the dorsal cochlear nucleus. PMID- 11474140 TI - Temporal relationship between the auditory brainstem response and focal responses of auditory nerve root and cochlear nucleus during development in the tammar wallaby (Macropus eugenii). AB - Thirty-two pouch-young tammar wallabies were used to discover the generators of the auditory brainstem response (ABR) during development by the use of simultaneous ABR and focal brainstem recordings. A click response from the auditory nerve root (ANR) in the wallaby was recorded from postnatal day (PND) 101, when no central auditory station was functional, and coincided with the ABR, a simple positive wave. The response of the cochlear nucleus (CN) was detected from PND 110, when the ABR had developed 1 positive and 1 negative peak. The dominant component of the focal ANR response, the N1 wave, coincided with the first half of the ABR P wave, and that of the focal CN response, the N1 wave, coincided with the later two thirds. In older animals, the ANR response coincided with the ABR's N1 wave, while the CN response coincided with the ABR's P2, N2 and P3 waves, with its contribution to the ABR P2 dominant. The protracted development of the marsupial auditory system which facilitated these correlations makes the tammar wallaby a particularly suitable model. PMID- 11474141 TI - A brief survey of the modifications in sensory-secretory organs of the neonatal rat tongue. AB - Recent data obtained on rats suggest that in the days immediately following birth several events take place in the circumvallate papillae of the oral cavity. A phylogenetically primitive system of solitary chemosensory cells develops and is rapidly replaced by taste buds. The lipase-secreting von Ebner gland, which is associated with taste organs, begins to develop by forming short tubules. The intrinsic nervous system of the gustatory organs rapidly completes its maturation showing fast proliferation of fibers and immunocytochemical maturation. Intraepithelial lipid accumulation is visible in the non-receptorial mucosa of the tongue, showing aspects which suggest an active lipid secretion. These data demonstrate that in the rat the structure of the sensory-secretory organs of the newborn's tongue shows a typical conformation with respect to the adult and rapidly changes its organization in the first week after the birth. At the present level of knowledge, it is difficult to link the anatomical structures to peculiar functional roles but the rather simple organization of the neonatal gustatory epithelium could be in relation to the dietary regimen. The data obtained in laboratory animals underline the necessity of studies on human newborns to update the anatomical knowledge of the oral chemoceptive system. PMID- 11474142 TI - Advanced magnetic resonance imaging techniques in perinatal brain injury. AB - Despite marked improvements in perinatal practice, perinatal brain injury remains one of the most common complications causing chronic handicapping conditions. Experimental advances have elucidated many of the cellular and vascular mechanisms of perinatal brain damage showing a correlation between the nature of the injury and the maturation of the brain. New diagnostic tools, such as quantitative three-dimensional magnetic resonance (MR) imaging, diffusion weighted MR imaging and proton MR spectroscopy, are presented in this review article that allow to assess brain development, detect early brain injury and monitor effects of perinatal brain injury on subsequent brain development and brain plasticity. These techniques will guide future therapeutic interventions aimed at minimizing irreversible perinatal brain injury. PMID- 11474143 TI - Sensorial saturation: an effective analgesic tool for heel-prick in preterm infants: a prospective randomized trial. AB - Pain is traumatic for preterm infants and can damage their CNS. We wanted to assess whether multisensorial stimulation can be analgesic and whether this effect is only due to oral glucose or sucking. We performed a randomized prospective study, using a validated acute pain rating scale to assess pain during heel-prick combined with five different procedures: (A) control, (B) 10% oral glucose plus sucking, (C) sensorial saturation (SS), (D) oral water, and (E) 10% oral glucose. SS is a multisensorial stimulation consisting of delicate tactile, vestibular, gustative, olfactory, auditory and visual stimuli. Controls did not receive any analgesia. We studied 85 heel-pricks (5 per baby) performed for routine blood samples in 17 preterm infants (28-35 weeks of gestational age). We applied in random order in each patient the five procedures described above and scored pain. SS and sucking plus oral glucose have the greater analgesic effect with respect to no intervention (p < 0.001). The effect of SS is statistically better than that of glucose plus sucking (p < 0.01). SS promotes interaction between nurse and infant and is a simple effective form of analgesia for the NICU. PMID- 11474144 TI - Fetal bile acid metabolism: analysis of urinary 3beta-monohydroxy-delta(5) bile acid in preterm infants. AB - To elucidate the urinary concentration of total bile acids after birth and the profile of the usual and unusual urinary bile acids, especially 3beta-hydroxy-5 cholen-24-oic acid (Delta(5)-3beta-ol), we measured the concentrations of 13 bile acids in the urine from preterm infants vs. full-term controls by gas chromatography-mass spectrometry. The urinary concentration of total bile acids in early preterm infants below 32 weeks of gestational age significantly exceeded that of the late preterm and full-term infants (p < 0.0005). The major urinary bile acids in early preterm infants were cholic acid, 1beta,3alpha,7alpha,12alpha tetrahydroxy-5beta-cholan-24-oic acid and Delta(5)-3beta-ol. In conclusion, the high urinary concentrations of total bile acids in preterm infants may be due to an overproduction, or more likely to a low hepatic bile acid clearance. An alternative fetal pathway, the acidic pathway, may be a major route of bile acid biosynthesis in preterm infants. PMID- 11474145 TI - Changes in the G gamma- and A gamma-globin mRNA components of fetal hemoglobin during human development. AB - To determine the postnatal developmental changes in (G)gamma- and (A)gamma-globin mRNAs of HbF, blood samples were obtained from 28 pre-term infants born at < or = 33 weeks of gestation (27.9 +/- 2.5), 9 term-born infants and 26 adults. Adult samples were used to determine the relationship between the levels (G)gamma- and (A)gamma-mRNAs of HbF and the-158 (C-->T) variation in the promoter region of the (G)gamma-globin gene. The results showed that (G)gamma-globin mRNA to total gamma globin mRNAs remained around 66% (66.2 +/- 4) until the 44 week of postconceptual age when a change in the (G)gamma- and (A)gamma-globin mRNA proportions occurred. The immature red cells of adults had a range of (G)gamma-globin mRNA to total gamma-globin mRNAs varying from 20 to 74% (53.1 +/- 16.9). The high levels of (G)gamma-globin mRNA to total gamma-globin mRNAs are associated with the presence of a -158 (C-->T) polymorphism. The gene frequency of polymorphism was 0.32. PMID- 11474146 TI - Effects of cisapride on ventricular depolarization-repolarization and arrhythmia markers in infants. AB - To study prospectively the effects of cisapride on ventricular repolarization, depolarization, and arrhythmia markers in neonates, we determined before and three days after starting cisapride (1 mg/kg/day): corrected QT interval (QTc) and QT dispersion (QTd) on standard ECGs, and duration of filtered QRS (fQRS) and of low amplitude (<40 microV) terminal signals (LAS40, ms) and root mean square of the last 40 ms (RMS40, microV) using high-gain signal-averaged ECG (SAECG). Twenty-four term and 11 preterm infants (gestational age 23-35 weeks) were studied at a median chronological age of 32 days. QTc and QTd were not different between term and preterm infants. Cisapride lengthened QTc (mean +/- SD; ms: 396.6 +/- 24.8 before vs. 417.0 +/- 35.2 after, p < 0.001). Three term and two preterm infants (5/35 = 14%; 95% CI: 5-30%) had a QTc >450 ms after cisapride. QTd after cisapride increased significantly in all infants with prolonged QTc. Filtered QRS, LAS40, and RMS40 before and after cisapride were within our normal values. We conclude that cisapride prolongs ventricular repolarization in neonates and infants without altering depolarization. Although no clinical arrhythmias were observed the dose of 0.8 mg/kg/day should not be exceeded. PMID- 11474147 TI - Immaturity or starvation? Longitudinal study of leptin levels in premature infants. AB - OBJECTIVE: Leptin, the protein product of the ob gene, is a potential placental growth factor and is integral to the body's system of energy regulation as shown in animal models. Premature infants are especially vulnerable to changes in energy regulation, and several studies have demonstrated a rapid fall in leptin values at birth. The purpose of the present investigation was to measure leptin levels in premature infants throughout hospitalization. METHODS: Eligible infants were less than 32 weeks' gestation, appropriate for gestational age, and hospitalized at Christiana Hospital Special Care Nursery. Serum samples for leptin analysis were drawn within 24 h of birth and twice a week thereafter until discharge. Concurrent growth measurements were obtained with each leptin sample. Body mass index, ponderal index, and midarm circumference/head circumference ratios were calculated to assess growth. RESULTS: Leptin levels were low and remained low for the duration of the premature infants' hospitalization (mean +/- SD = 1.35 +/- 0.63 ng/ml/ml, range 0-3.06). After controlling for weight, there was a small (r(2) = 0.1, p < 0.00001) but significant correlation between leptin and postnatal age after 4 days of age. Despite an increase in caloric intake during the study period, there was no relationship between leptin and caloric intake. There were significant negative correlations between measurements of growth and both leptin and the leptin/weight ratio. Maternal diabetes and the use of steroids had small but significant effects on the leptin/weight ratio. CONCLUSION: In this population of predominantly female premature infants, leptin levels were very low as compared to term infants, children and adults, and did not change appreciably over the study period. The low leptin levels seen in these premature infants are similar to those levels seen in malnourished adults, anorexics, and in animal models of starvation. We speculate that a critical adipose store needs to be reached before increased amounts of leptin can be adequately produced. Persistently low leptin levels may also reflect an immaturity in the hypothalamic-pituitary-adrenal axis. PMID- 11474148 TI - Inflammatory mediators in umbilical plasma from neonates who develop early-onset sepsis. AB - OBJECTIVES: To study whether early-onset neonatal sepsis is associated with a prenatal immune response with elevated umbilical plasma levels of inflammatory mediators, and to study whether mediator levels may be helpful in identifying infected neonates. SETTING: Nested case-control study. METHODS: Cord blood was sampled from 7,073 consecutively delivered neonates. After review of the medical records, neonates suspected to suffer from infection were classified as infected (n = 52) or noninfected but sick controls (n = 33). We also included a group of healthy controls (n = 99). Umbilical plasma levels of tumour necrosis factor alpha (TNFalpha), interleukin (IL)-1beta, IL-6, IL-8, soluble TNF receptors (p55 and p75), IL-1 receptor antagonist (IL-1RA) and C-reactive protein were measured by immunoassays. RESULTS: Infected neonates had higher levels of TNFalpha, IL 1beta, IL-6, IL-8, p55, p75 and IL-1RA than healthy controls (all p < 0.01). Among preterm infants (GA <37 weeks), those with infection (n = 11) had higher levels of IL-1beta, IL-6, IL-8, p55 and p75 than noninfected sick controls (n = 13) (all p < 0.05), but among term infants, the infected did not differ from the noninfected sick controls. Receiver operator characteristic plots showed that IL 1beta, IL-6 and IL-8 identified preterm infected neonates accurately. CONCLUSIONS: Early-onset neonatal sepsis is associated with a prenatal immune response with increased TNFalpha, IL-1beta, IL-6, IL-8, p55, p75 and IL-1RA levels in umbilical plasma. Among neonates who present symptoms suggestive of infection, cytokine levels may be helpful in identifying preterm, but not term infected individuals. PMID- 11474149 TI - Estimation of mean nuclear volume of neocortical neurons in sudden infant death syndrome cases using the nucleator estimator technique. AB - At present, no information is available with regards to either neocortical neuronal mean nuclear volume or maturation (functional or morphological) in abnormal paediatric brains. Using the nucleator estimator technique, the mean neocortical nuclear volume was estimated in sudden infant death syndrome (SIDS) cases [10 normal birth weight (NBW) and 10 low birth weight (LBW) cases classified by birth weight for gestational age] and compared to 10 NBW control cases. Both the control and SIDS LBW cases showed an increase in mean nuclear volume with age; the SIDS NBW cases showed no increase. At 8 months, the SIDS NBW cases showed a reduced mean nuclear volume (p = 0.02) when compared to controls. The SIDS LBW cases showed no statistically significant difference (p = 0.10) when compared with controls. A deficiency in mean neuronal nuclear volume may represent a deficiency in neuronal function. PMID- 11474150 TI - 7-Nitroindazole, but not aminoguanidine, attenuates the acute inflammatory responses and brain injury during the early phase of Escherichia coli meningitis in the newborn piglet. AB - We evaluated the anti-inflammatory and neuroprotective effects of the selective neuronal nitric oxide synthase inhibitor 7-nitroindazole and aminoguanidine, which predominantly inhibits inducible nitric oxide synthase, during the early phase of experimental bacterial meningitis in the newborn piglet. Meningitis was induced by intracisternal injection of 10(8) colony-forming units of Escherichia coli in 100 microl of saline. 7-Nitroindazole significantly attenuated the meningitis-induced acute inflammatory responses such as increased intracranial pressure, decreased cerebrospinal fluid (CSF) glucose concentration, and CSF leukocytosis at 2 h. However, meningitis-induced CSF leukocytosis at 4 h and increased CSF lactate and tumor necrosis factor alpha levels were not significantly attenuated. Reduced cerebral cortical cell membrane Na(+),K(+) ATPase activity and increased lipid peroxidation products, indicative of meningitis-induced brain cell membrane dysfunction, were also significantly improved with 7-nitroindazole treatment. In contrast, although aminoguanidine significantly attenuated the increase in the CSF tumor necrosis factor alpha level, it failed to attenuate the acute inflammation and the ensuing brain injury in bacterial meningitis. In summary, 7-nitroindazole, but not aminoguanidine, significantly attenuated the acute inflammatory responses and brain injury during the early phase of neonatal bacterial meningitis. PMID- 11474151 TI - Effect of N(G)-nitro-L-arginine methyl ester on intestinal permeability following intestinal ischemia-reperfusion injury in a rat model. AB - Subclinical intestinal ischemia-reperfusion injury (IRI) causes an increase in mucosal permeability and may represent an early event in the pathogenesis of necrotizing enterocolitis in premature infants. Previous studies suggested that continuous, endogenous formation of nitric oxide (NO) maintains the mucosal integrity of the intestine, thus protecting the gut from injuries from blood borne toxins and tissue-destructive mediators. This study was undertaken to assess whether the inhibition of NO production causes an increase in intestinal permeability in rats following IRI. Sprague-Dawley rats weighing 200-300 g were divided into 4 groups: (1) untreated group (normal control); (2) ischemia reperfusion group; (3) early N(G)-nitro-L-arginine methyl ester (L-NAME), a specific inhibitor of NO production, treatment group, and (4) late L-NAME treatment group. Transient IRI was induced by 30-min occlusion, followed by reperfusion of the isolated ileal loop. The L-NAME was administered 15 min before and after mesenteric ischemia as a 25-mg/kg bolus. Fluorescein isothiocyanate dextran (FITC-D) was used to quantitatively assess the alteration in mucosal permeability of the intestine. There was no significant increase in the portal vein FITC-D level among normal controls, ischemia-reperfusion group and late L NAME-treated group, but there was an approximately 6-fold increase in the early L NAME treatment group. The pathological features of the intestine following IRI include denudation of the villus epithelium and reduction of villus height, associated with marked inflammatory cell infiltration over the lamina propria. These results suggest that endogenous NO may play a role in the protecting intestinal integrity after IRI. PMID- 11474152 TI - Increased constriction of the ductus arteriosus with combined administration of indomethacin and L-NAME in fetal rats. AB - We studied age-related changes and the caliber of the ductus arteriosus (DA) after two-pathway inhibition of prostaglandin E(2) and nitric oxide (NO) by the combined administration of indomethacin, a cyclooxygenase inhibitor, and N(G) nitro-L-arginine methyl ester (L-NAME), an NO synthase inhibitor, in fetal rats. Pregnant rats from day 18 to 21 of gestation were used. They were administered indomethacin orally (3 mg/kg) 3 h before cesarean section, and then L-NAME (50 mg/kg) was injected intraperitoneally 3 h before the rats were killed. Using rapid-freezing and shaving methods, the caliber of the DA in fetal rats was measured. Compared with the indomethacin alone group, indomethacin plus L-NAME further constricted the DA after indomethacin and L-NAME were simultaneously administered 3 h before the rats were sacrificed. The extent of the final DA constriction was almost equal to the addition of each effect of indomethacin and L-NAME. We concluded that the magnitude of DA constriction following indomethacin plus L-NAME was due to the additive effects of these agents, suggesting a possible method to treat patent DA in premature infants. PMID- 11474153 TI - mRNA D(2) dopaminergic receptor expression after hypoxia-ischemia in rat immature brain. AB - Previous studies have shown a reduction of dopaminergic D(2) receptors (D(2)R) in the striatum after hypoxia-ischemia in newborn rats. We show here an early and transient reduction of mRNA D(2)R in nonatrophic brains following hypoxia ischemia. The left carotid artery of P7 rats was ligated followed by hypoxia for 2 h. The rats were sacrificed after 24 h, 48 h and 14 days. D(2)R mRNA was studied by in situ hybridization, the cell number by conventional histology, and neuronal and astrocyte differentiation by immunohistochemistry. A 20% reduction of striatal mRNA D(2)R occurred 24 h after hypoxia-ischemia, whereas no reduction was observed after 48 h and 14 days. There were no differences in total cell number and in the expression of neuronal (MAP-1, MAP-2) and astrocyte (GFAP) markers between both brain hemispheres nor between control and hypoxia-ischemia animals. The early decrease in mRNA D(2)R could explain the delayed reduced D(2)R after neonatal hypoxia-ischemia. PMID- 11474154 TI - Hemodynamic effects of levcromakalim in neonatal porcine pulmonary hypertension. AB - Levcromakalim (LKM; a K(ATP) channel opener) reverses hypoxic pulmonary vasoconstriction in isolated pulmonary arteries and perfused lungs. This vasorelaxation is blocked by glibenclamide (GLB; a K(ATP) channel blocker). We evaluated the hemodynamic effect of LKM followed by GLB in a chronically instrumented neonatal porcine model of pulmonary hypertension, created by exposing piglets to hypoxia (n = 7) or heat-killed group B streptococci (GBS) (n = 6). Hypoxia increased pulmonary arterial pressure (PAP), which LKM decreased, and GLB subsequently increased in a dose-dependent manner. Systemic arterial pressure (SAP) did not change with hypoxia but was also decreased by LKM and increased by GLB. GBS also led to increased PAP, but LKM significantly reduced only SAP, which was then increased by GLB. We conclude LKM is capable of reversing hypoxic, but not GBS-induced, pulmonary hypertension but lacks specificity for the neonatal pulmonary vasculature. PMID- 11474155 TI - Prenatal influence of ischemia-hypoxia-induced intrauterine growth retardation on brain development and behavioral activity in rats. AB - The effects of intrauterine growth retardation (IUGR) on brain histological or functional development were examined in rats. IUGR was induced by ligating the bilateral uterine arteries at day 17 of pregnancy. On day 22 of pregnancy, cesarean section was performed, and pups with a birth weight of <2 SD of the mean birth weight of control pups were regarded as IUGR rats. Morphological changes of the brain were studied by Nissl's staining at different timepoints during prenatal and postnatal periods. For behavioral study, an open-field test was performed at 5, 7 and 10 weeks after birth. Histological studies showed the migration disorder of the neurons in the cerebral cortex from embryonic day 17 to postnatal day (PD) 49. The open-field test revealed locomotor disturbance at PD49 in male IUGR rats, but not in female IUGR rats or control rats. It is concluded that IUGR due to antenatal ischemia-hypoxia causes morphological changes in the central nervous system, and induces behavioral impairment, particularly in male rats. PMID- 11474156 TI - Aspirin and percutaneous coronary angioplasty are associated with a decline in mortality from cardiogenic shock. Results from a national Israeli survey, 1992- 1998. AB - BACKGROUND AND METHODS: Despite the significant progress in the care and outcome of patients with acute myocardial infarction (AMI), the impact of evolving therapies on the incidence and outcome of patients with cardiogenic shock complicating AMI has been questioned. We analyzed trends in the incidence, care and outcome of cardiogenic shock from four national surveys conducted during 1992 -1998. RESULTS: Of the 5,351 AMI patients admitted to all coronary care units in Israel, 254 (4.7%) developed cardiogenic shock. The incidence of cardiogenic shock decreased over time (5.8, 5.1, 4.3 and 4.4% for the years 1992, 1994, 1996 and 1998, respectively, p = 0.08). Concomitantly, there was an increase in utilization of coronary angiography, urgent angioplasty and intra-aortic balloon counterpulsation. In addition, there was an increase in hospital use of aspirin, nitrates, ACE inhibitors and beta-blockers. Patients with shock were more likely to die within 7 days compared with AMI patients not having shock (65 vs. 4%; p < 0.001). During the study period, the mortality of patients with shock decreased: at 7 days (72% in 1992 to 60% in 1998; p = 0.09), at 30 days (87 to 70%, respectively; p = 0.01) and at 6 months (89 to 77%, respectively; p = 0.02). Both aspirin and angioplasty were independently associated with improved outcome after adjustment for baseline characteristics and study period. CONCLUSIONS: Although the mortality rate of cardiogenic shock complicating AMI remains high, the increased utilization of aspirin and angioplasty is associated with improved outcome. PMID- 11474157 TI - The peripheral vascular response to exercise is impaired in patients with risk factors for coronary artery disease. AB - BACKGROUND: Abnormal coronary and brachial artery responses have been described in individuals with risk factors for coronary artery disease (CAD). Peripheral arterial tonometry (PAT), a newly developed digital plethysmographic technique was used to assess peripheral vascular response to exercise in healthy controls and individuals with risk factors. METHODS AND RESULTS: Continuous finger PAT during Bruce protocol exercise test was performed in 30 subjects with risk factors for CAD and 30 healthy individuals. Compared with baseline, the PAT wave amplitude at peak exercise decreased in the subjects but increased in the controls: 83 +/- 28% vs. 114 +/- 40% respectively, p < 0.01. CONCLUSIONS: A different pattern of systemic vascular response to exercise was found in individuals with risk factors for atherosclerosis. Since the vascular behavior in these patients is probably related to endothelial dysfunction, it may be that peripheral arterial tonometry can be used as a simple, readily available technique to assess endothelial function. PMID- 11474158 TI - Determinants of short-period heart rate variability in the general population. AB - Decreased heart rate variability (HRV) is associated with a worse prognosis in a variety of diseases and disorders. We evaluated the determinants of short-period HRV in a random sample of 149 middle-aged men and 137 women from the general population. Spectral analysis was used to compute low-frequency (LF), high frequency (HF) and total-frequency power. HRV showed a strong inverse association with age and heart rate in both sexes with a more pronounced effect of heart rate on HRV in women. Age and heart rate-adjusted LF was significantly higher in men and HF higher in women. Significant negative correlations of BMI, triglycerides, insulin and positive correlations of HDL cholesterol with LF and total power occurred only in men. In multivariate analyses, heart rate and age persisted as prominent independent predictors of HRV. In addition, BMI was strongly negatively associated with LF in men but not in women. We conclude that the more pronounced vagal influence in cardiac regulation in middle-aged women and the gender different influence of heart rate and metabolic factors on HRV may help to explain the lower susceptibility of women for cardiac arrhythmias. PMID- 11474159 TI - Interpretation of high wedge pressure on exercise in patients with chronic obstructive pulmonary disease. AB - In a retrospective review of patients subjected to right heart catheterization, case records from 109 consecutive patients with chronic obstructive pulmonary disease (COPD) and wedge pressure > or =15 mm Hg on exercise were analyzed. Patients were separated into group 1, resting wedge pressure (P macro w) <15 mm Hg and difference between P macro w and right atrial pressure (P macro ra) change on exercise <5 mm Hg (n = 54), group 2, same P macro w at rest but increase in macro on exercise > or =5 mm Hg, being higher than that of P macro ra (n = 34), and group 3, P macro w > or =15 mm Hg at rest (n = 21). The occurrence of left heart disease increased from group 1 to group 3 (19, 53 and 71%, respectively), and the slope of the P macro w/stroke index relationship was lowest in group 3. High P macro w on exercise can be explained by the pressure rise in the cardiac fossa associated with lower lobe gas trapping in group 1, which showed the most severe airflow limitation, decreased left ventricular compliance in group 2, and heart failure in group 3. PMID- 11474160 TI - Efficacy and safety of imidapril in patients with essential hypertension: a double-blind comparison with captopril. AB - In this 12-week, double-blind, parallel-group, comparative trial, 57 adult patients with mild-to-moderate hypertension were randomly allocated to receive imidapril or captopril, initially at a dose of 5 mg once a day and 25 mg twice daily, respectively. After 4 weeks of therapy, the dose of each drug was increased twice if diastolic blood pressure (DBP) remained > or =90 mm Hg. Both treatments effectively lowered DBP in a comparable manner. Mean changes from baseline in DBP at 12 weeks were -9.9 mm Hg for imidapril and -8.8 mm Hg for captopril (p = 0.488). Responder rates in patients receiving active treatment for at least 6 weeks were 53.9% for imidapril and 48% for captopril (p = 0.676). Both treatments were well tolerated. Adverse drug reactions were observed in 20.7% (6/29) of the imidapril group and 46.4% (13/28) of the captopril group (p < 0.05). A cough was the most frequent side effect, reported in 13.8% of the imidapril group and 35.7% of the captopril group. The results indicate that imidapril is as effective as captopril in the treatment of hypertension. Imidapril produces less adverse effects compared with captopril. PMID- 11474161 TI - Role of intraventricular dispersion of early diastolic filling in indicating left ventricular diastolic dysfunction: assessment by color M-mode inflow propagation velocity. AB - The intraventricular to mitral E velocity ratio (IvE/MvE) and the color M-mode Doppler inflow propagation velocity (LVIPV) were evaluated in 36 healthy controls and 33 patients with hypertension and/or ischemic heart disease. The intraventricular E velocity was significantly lower than the mitral E velocity in the control group (52 vs. 68 cm/s, p < 0.001), but they are similar in the disease group (59 vs. 63 cm/s, p = nonsignificant). Compared with the control group, the disease group had a higher IvE/MvE (0.95 +/- 0.28 vs. 0.78 +/- 0.23, p < 0.01). Nevertheless, the LVIPV was not significantly different between the disease and control groups (53 +/- 14 vs. 56 +/- 13 cm/s, p = nonsignificant). The LVIPV did not correlate with the IvE/MvE (r = 0.202, p = nonsignificant). Therefore, an intraventricular dispersion of early diastolic filling does not seem to indicate impaired left ventricular relaxation. PMID- 11474162 TI - Can pre-treatment with beta-agonists reduce stress test time and the use of atropine in dobutamine stress testing? AB - AIM: To determine whether the use of inhaled beta-agonists decreases the duration of dobutamine stress testing (DAS), reduces the amount of dobutamine infused and decreases the use of atropine. METHODS: 34 patients on beta-agonists (group A) (mean age 65 +/- 8 years) and 32 patients not on beta-agonists or beta-blockers (group B) (mean age 65 +/- 10 years) undergoing DAS with SPECT were enrolled. RESULTS: The time of infusion of dobutamine in group A was 6.41 +/- 1.58 min and in group B was 9.77 +/- 3.60 min (p < 0.001). The amount of dobutamine infused was 10.64 +/- 5 mg in group A and 19.20 +/- 8 mg in group B (p < 0.001). In group A, 2 of 34 patients and 18 of 32 patients in group B needed atropine to reach peak HR (p < 0.001). CONCLUSION: Patients on short acting beta-agonists require smaller amounts of dobutamine with a shorter infusion time during DAS, and lesser use of side-effect prone atropine. PMID- 11474163 TI - Comparative evaluation of thermodilution and gated blood pool method for determination of right ventricular ejection fraction at rest and during exercise. AB - BACKGROUND: Since the development of a Swan-Ganz thermodilution ejection fraction catheter several studies have been published which compare this technique for obtaining right ventricular ejection fraction (RVEF(TD)) with alternative methods. However, the reliability of RVEF(TD) measurements under exercise conditions remains undetermined. Therefore, the aim of the present study was a comparative evaluation of RVEF(TD) with the established gated blood pool method (RVEF(GBP)) under exercise conditions. METHODS AND RESULTS: Twenty-two patients with different cardiac diseases underwent right heart catheterization, including RVEF(TD) and simultaneous RVEF(GBP) determination at rest and during supine bicycle exercise. Linear regression analysis showed a significant correlation between RVEF(TD) and RVEF(GBP) at rest (r = 0.73, p < or = 0.0005) and during exercise (r = 0.74, p < or = 0.0005). A Wilcoxon analysis showed a high probability of agreement of RVEF(TD) and RVEF(GBP) at rest and exercise (level of significance for error of the 0 hypothesis of 95.9/73.3%). CONCLUSION: The thermodilution ejection fraction catheter provides a useful device for reliable, repetitive and safe RVEF measurements, not only at rest but also under exercise conditions. This seems to be clinically important, because by it means RVEF, as a sensitive parameter of primary or secondary right ventricular dysfunction, can be determined in the course of standard right heart catheterization. PMID- 11474164 TI - Pulsation, systolic thrill and murmur in femoral veins secondary to severe tricuspid regurgitation. AB - We report on a case of pulsatile femoral veins with a systolic thrill and murmur in a 26-year-old patient who exhibited severe tricuspid regurgitation. The pulsatile nature of the veins may result from the 'ventricularization' of venous pressure with each pressure pulse. The observed systolic thrill and murmur may be due to the systolic reversal of substantial regurgitant flow in the venous system of the lower limbs. This case also demonstrates that severe tricuspid regurgitation can have far-reaching manifestations. PMID- 11474165 TI - Arrhythmias in neurofibromatosis. A case report and review of the literature. AB - Patients with neurofibromatosis have a higher incidence of anatomic cardiac abnormalities. However, there is little data regarding incidence of arrhythmias in this population. It is known that these patients have a higher mortality than the normal population, and it is possible that some deaths may be due to preventable causes such as cardiac arrhythmias. We report a patient with neurofibromatosis who was treated for a refractory seizure disorder for 8 years. However, video/EEG monitoring demonstrated that the patient had recurrent syncopal seizures secondary to sinus node dysfunction. Complete resolution of symptoms occurred after a permanent pacemaker implantation. We believe this is the first reported case of sinus node dysfunction associated with neurofibromatosis. PMID- 11474166 TI - False-positive troponin I in a patient with acute cholecystitis and positive rheumatoid factor assay. PMID- 11474167 TI - Point-of-care monitoring of patients with left ventricular dysfunction on cardiovascular drugs. PMID- 11474168 TI - Classifying by colors: FISH-based genome analysis. AB - In recent years a fascinating evolution of different multicolor fluorescence in situ hybridization (FISH) technologies could be witnessed. The various approaches to cohybridize multiple DNA probes in different colors opened new avenues for FISH-based automated karyotyping or the simultaneous analysis of multiple defined regions within the genome. These developments had a remarkable impact on microscopy design and the usage of highly sensitive area imagers. In addition, they led to the introduction of new fluorochromes with appropriate filter combinations, refinements of hybridization protocols, novel probe sets, and innovative software for automated chromosome analysis. This paper attempts to summarize the various multicolor approaches and discusses the application of the individual technologies. PMID- 11474169 TI - Facilitating haplotype analysis by fully automated analysis of all chromosomes in human-mouse hybrid cell lines. AB - Recent evidence suggests that haplotype analysis is essential in recognizing genetic factors involved in the tendency toward a particular disease or pharmacogenetic phenotype, as well as to identify genes involved in multigenic disorders. Because of the increasing need for efficient haplotype tests, a new hybrid system, called conversion technology, was developed. Conversion technology aims at converting the diploid chromosome content into a haploid state so that hybrids contain a single copy of any desired chromosome. A number of mutations can now be identified easily, as they are no longer obscured by the normal sequence present on the other copy of the chromosome. However, the efficient use of this hybrid system depends on a complete analysis of both human and mouse chromosome complements in order to assess the stability of the hybrid cells and to accurately determine their human chromosome content. We describe a new multicolor FISH-based method capable of analyzing both genomes simultaneously in a single hybridization. This new technique should become an instrumental part of inexpensive, reliable haplotype tests. PMID- 11474170 TI - Use of primed in situ labeling (PRINS) for the detection of telomeric deletions associated with mental retardation. AB - Rearrangements involving the telomeric regions of human chromosomes are often associated with mental retardation. These rearrangements, however, are difficult to detect using conventional cytogenetic techniques. We propose the use of primed in situ (PRINS) labeling as an alternative to fluorescence in situ hybridization because it is very fast, reproducible, and simple to perform. Sixty-five children with unexplained mental retardation were studied using PRINS technology; two of them were shown to have a telomeric deletion. PMID- 11474171 TI - C18orf2, a novel, highly conserved intronless gene within intron 5 of the GNAL gene on chromosome 18p11. AB - We have characterized a novel intronless human gene (C18orf2) which is embedded in intron 5 of the G-protein gene (GNAL) on chromosome 18p11. This gene codes for a 199 amino acid polypeptide with a predicted molecular weight of 22.1 kDa. It is highly homologous to a number of predicted developmental proteins in organisms ranging from yeasts to Drosophila. C18orf2 mRNA was found to be expressed in various tissues. PMID- 11474172 TI - Molecular cloning of a human novel type I cytokine receptor related to delta1/TSLPR. AB - In a search for a human sequence related to a recently identified type I cytokine receptor delta1, which turned out to be a receptor subunit for a cytokine called TSLP, we have now identified a novel human type I cytokine receptor from a human T lymphocyte cDNA library. The deduced amino acid sequence of 371 residues has a typical signal sequence and a membrane-spanning region. The mature protein is predicted to have a molecular mass of 39,698 Da. The N-terminal extracellular region contains two fibronectin type III-like domains, four conserved cysteine residues, and a WSXWS box-like motif. The C-terminal intracellular region contains box 1 and box 2-like motifs. Thus, it has common characteristics of type I cytokine receptor family members, and we tentatively termed this protein CRLF2, which stands for cytokine receptor-like factor 2. Northern blot analysis revealed CRLF2 mRNA in liver, kidney, heart, and skeletal muscle. The fetal liver also expresses CRLF2 transcripts. The gene for CRLF2 was mapped to the pseudoautosomal region, Xp22.3 and Yp11.3 by FISH analysis, a region where genes encoding the IL 3 receptor alpha and the GM-CSF receptor alpha chains are also located. The biological function of this newly identified receptor is now under investigation. PMID- 11474173 TI - Genomic structure, chromosome mapping and expression analysis of the human AXIN2 gene. AB - Conductin is a Wnt signalling protein and serves as a negative regulator of beta catenin stability. We have previously isolated the human homolog (AXIN2) of the murine conductin gene and shown that it is mutated in colorectal cancer (CRC) with defective mismatch repair (MMR). Here we report the detailed genomic structure of this gene by analysis of cDNA and genomic clones. The gene spans > or =25 kb containing ten exons ranging from 96 bp to 904 bp. All splice donor and acceptor sites conform to the GT/AG rule. FISH (Fluorescence in situ Hybridization) analysis localized this gene to human chromosome band 17q24 and showed that it exists as a single copy in the human genome. Northern blot analysis from different human organs demonstrated that the AXIN2 gene is highly expressed in human thymus, prostate, testis, small intestine and ovarian tissues but expressed at a lower level in colon. The data reported here provides a framework for further analysis of this important Wnt signalling protein in vertebrate development and tumorigenesis. PMID- 11474174 TI - Fine-mapping of cytogenetically undetectable EWS/ERG fusions on DNA fibers of Ewing tumors. AB - In contrast to the EWS/FLI1 fusion which is represented by a t(11;22)(q24;q12), EWS/ERG fusions are frequently cytogenetically not detectable. Three Ewing tumors (ET), two with apparently normal chromosomes 21 and 22, and one ET with a t(2;22)(p25;q12), were studied by FISH on interphase nuclei, metaphase chromosomes and on DNA fibers. EWS/ERG transcripts were detected by RT-PCR in all cases. FISH, using cosmids located proximally (F10, G9) and distally (F7) to the EWS breakpoint region, revealed no detectable separation of these probes in two cases. In contrast, co-hybridization of probe PT1526 containing the ERG breakpoint region with G9 revealed the juxtaposition of two signals per interphase nucleus in all three cases indicating the EWS/ERG fusions. Chromosome preparations displayed the juxtaposed signals on the der(22), and hybridization signals of the probes PT1526 and G9 on the non-rearranged chromosomes 21 and 22 in all cases, respectively. The PT1526 signal on the der(21) was seen only in cases 1 and 2. These results were confirmed by triple-target FISH on tumor DNA fibers. In all three cases, the hybridization pattern F10 - G9 - PT1526 indicates a centromere to telomere orientation. This finding suggests that EWS/ERG fusions in ETs may be generated by an inversion of the ERG gene or a part thereof followed by an insertion into the EWS gene on the der(22). Double-target FISH on interphase nuclei using probes flanking the EWS breakpoint region and probe PT1526 enables the detection of virtually all 22q12 rearrangements in ETs, thus providing a reliable diagnostic assay. PMID- 11474175 TI - Isolation, characterization, and chromosome mapping of a human A-C1 Ha-Ras suppressor gene (HRASLS). AB - Recently, we cloned a cDNA encoding a novel mouse protein, named A-C1, by differential display between two mouse cell lines, embryonic fibroblast C3H10T1/2 and chondrogenic ATDC5. Mouse A-C1 has homology with a ras-responsive gene, rat Ha-rev107 (Hrasls), and modulates a Ha-ras-mediated signaling pathway. Here, we report a cDNA encoding a human homolog of mouse A-C1. The deduced amino acid sequence of human A-C1 consists of 168 amino acids, and shows 83% identity with that of mouse A-C1. Human A-C1 mRNA was expressed in skeletal muscle, testis, heart, brain, and thyroid in vivo. Moreover, expression of human A-C1 mRNA was detected at a high level in human osteosarcoma-derived U2OS cells in vitro. By FISH analysis the human A-C1 gene (HRASLS) was mapped to human chromosome 3q28--> q29. PMID- 11474176 TI - Genomic organization and mapping of the gene (SLC25A19) encoding the human mitochondrial deoxynucleotide carrier (DNC). AB - The deoxynucleotide carrier (DNC) transports deoxynucleotides into mitochondria and is therefore essential for mtDNA synthesis. The human DNC gene (SLC25A19) spans about 16.5 kb and consists of nine exons with the translation start site in exon 4. It is located on chromosome 17q25.3. Three transcripts, which differ in their 5' ends and are generated by alternative splicing, have been identified. PMID- 11474177 TI - A physical and expression map of the D17S1810-D17S1353 region spanning the central areolar choroidal dystrophy locus. AB - Central areolar choroidal dystrophy (CACD) causes bilateral irreversible central visual loss in the 5th to 7th decades. The authors previously described a large pedigree with the disorder, which showed linkage to chromosome 17p13.2-->p13.1 between microsatellite markers D17S1353 and D17S1810. 17p13 is very rich in genes that cause retinal diseases. We have now constructed a detailed and ordered physical map of the critical CACD region which spans up to 2.4 Mb. The new transcript map contains thirteen genes and seven expressed sequence tags (ESTs) that are eye-expressed, and therefore are positional candidates. Several of these have been screened, but no disease-causing mutations were found in CACD patients. PMID- 11474178 TI - Delineation of a 150-kb breakpoint cluster in benign thyroid tumors with 19q13.4 aberrations. AB - Structural rearrangements involving the long arm of chromosome 19 characterize a cytogenetic subgroup of benign thyroid tumors and constitute one of the most frequent specific chromosome abnormalities in epithelial tumors. Recently, we have been able to narrow down the breakpoint region affected in two cell lines to a region covered by a single PAC clone. Close to that region a candidate gene has been identified which we tentatively referred to as RITA (Rearranged In Thyroid Adenomas) now named ZNF331 according to HUGO nomenclature. However, the results had been obtained on two cell lines only making it necessary to extend the studies to a larger number of tumors including primary material. Herein, we have used four further primary tumors showing translocations involving 19q13 for fluorescence in situ hybridization (FISH) mapping studies using a variety of molecular probes from a 470-kbp cosmid/BAC contig. Ten new STSs were characterized and physically mapped within an EcoRI restriction map. The results enabled us to define an approximately 150-kbp breakpoint cluster region of the 19q13 aberrations in benign thyroid tumors flanked by two newly established STS markers. PMID- 11474179 TI - Expression and chromosome location of hamster Ku70 and Ku80. AB - Ku proteins play an important role in DNA double-strand break (DSB) repair, chromosome maintenance, and growth regulation. To understand the fundamental characteristics of Ku proteins, we examined the electrophoretic mobility and expression of hamster Ku70 and Ku80 and determined the chromosome locations of their genes. The electrophoretic mobility of hamster Ku proteins are different from that of human Ku proteins. No significant changes in the quantity of Ku proteins were observed in CHO-K1 cells treated with 10 Gy of ionizing radiation, suggesting that both proteins are expressed constitutively in amounts adequate to repair DNA DSBs. The chromosome locations of the Ku genes were determined by direct R-banding fluorescence in situ hybridization. The Ku70 gene was localized to Syrian hamster chromosome 4qa4.1--> qa4.2 and Chinese hamster chromosome 2p3.1, and the Ku80 gene was localized to Syrian hamster chromosome 4qb5--> qb6.1 and Chinese hamster chromosome 2p3.5-->p3.6. These results provide clues to the biological functions of Ku, as well as useful information for constructing comparative chromosome maps between hamsters and other mammalian species, including human, mouse, and rat. PMID- 11474180 TI - Chromosomal and comparative mapping of rat oxytocin, oxytocin receptor and vasopressin genes. AB - Oxytocin and its receptor are potentially important for cardiovascular functions. In the present paper, we report their chromosome locations in the rat and their comparative mapping with the mouse and human. They are located in chromosome regions previously known to contain quantitative trait loci for blood pressure in various genetic crosses. Thus, they have become valid candidate genes for genetic hypertension. PMID- 11474181 TI - Assignment of eight additional genes from human chromosome 11 to bovine chromosomes 15 and 29: refinement of the comparative map. AB - A comparative mapping approach was applied in order to refine the extent and the distribution of conserved segments between human chromosome 11 (HSA11) and cattle chromosomes 15 and 29 (BTA15 and BTA29 respectively). Eight genes from HSA11 were mapped using a bovine-hamster somatic cell hybrid panel and seven represent new assignments. Adding these assignments to those present in human, mouse and cattle databases, a new conserved segment was identified between the telomeric region of HSA11 and BTA29. This brings to seven the number of conserved segments identified between HSA11 and BTA15 and 29, and our study refines their boundaries to the level of the human cytogenetic band. PMID- 11474182 TI - Genomic organization and assignment of the interleukin 7 gene (IL7) to porcine chromosome 4q11-->q13 by FISH and by analysis of radiation hybrid panels. AB - Interleukin 7 (IL7) is a cytokine that has many immunological functions, including regulation of hematopoiesis and peripheral lymphocytes. cDNA and a genomic DNA segment containing the porcine IL7 gene were isolated and sequenced, showing that porcine IL7 consists of 176 amino acids and that its gene spans over about 13 kb of genomic DNA. Porcine IL7 has 85% and 73% homology with human IL7 in terms of the nucleotide and amino acid sequences, respectively. Whereas the murine IL7 gene does not have an exon corresponding to human exon 5 (Lupton et al., 1990), the porcine IL7 gene was found to contain the same exon-intron structure as the human gene. These findings, together with the upstream structure of the cDNA elucidated in the present study, indicate that the relationship between swine and human IL7 is closer than that between mouse and human IL7. The IL7 gene was mapped to swine chromosome 4q11-->q13 by fluorescence in situ hybridization and, using a radiation hybrid panel, was localized between microsatellite markers Sw1336 and Sw1073 on the same chromosome. PMID- 11474183 TI - DNA sequence and physical mapping of the canine transglutaminase 1 gene. AB - The transglutaminase 1 gene (TGM1) encodes an enzyme necessary for cross-linking the structural proteins that form the cornified envelope, an essential component of the outermost layer of the skin, the stratum corneum. Reported here is the complete coding region of canine TGM1, its chromosome localization, and its map position in the integrated canine linkage-radiation hybrid map. Canine TGM1 consists of 2,448 nucleotides distributed over 15 exons. The nucleotide sequence has 90% identity to human TGM1. The deduced canine TGM1 protein is 816 amino acids long and is 92% identical to human TGM1. Using fluorescence in situ hybridization, we localized canine TGM1 to dog (Canis familiaris) chromosome 8 (CFA 8q). Canine TGM1 localized to CFA 8 on the integrated linkage-radiation hybrid map in the interval FH2149-MYH7. Characterizing the coding region of canine TGM1 is a first step in examining the role of this enzyme in normal and defective cornification in the dog. PMID- 11474184 TI - Mouse paracentric inversion In(3)55Rk mutates the urate oxidase gene. AB - The paracentric inversion In(3)55Rk on mouse Chromosome 3 (Chr 3) was induced by cesium irradiation. Genetic crosses indicate the proximal breakpoint cosegregates with D3Mit324 and D3Mit92; the distal breakpoint cosegregates with D3Mit127, D3Mit160, and D3Mit200. Giemsa-banded chromosomes show the inversion spans approximately 80% of Chr 3. The proximal breakpoint occurs within band 3A2, not 3B as reported previously; the distal breakpoint occurs within band 3H3. Mice homozygous for the inversion exhibit nephropathy indicative of uricase deficiency. Southern blot analyses of urate oxidase, Uox, show two RFLPs of genomic mutant DNA: an EcoRI site between exons 4-8 and a BamHI site 3' to exon 6. Mutant cDNA fails to amplify downstream of base 844 at the 3' end of exon 7. FISH analysis of chromosomes from inversion heterozygotes, using a cosmid clone containing genomic wild-type DNA for Uox exons 2-4, shows that a 5' segment of the mutated Uox allele on the inverted chromosome has been transposed from the distal breakpoint region to the proximal breakpoint region. Clinical, histopathological, and Northern analyses indicate that our radiation-induced mutation, uox(In), is a putative null. PMID- 11474185 TI - Mapping of the rat glypican genes. AB - The glypicans compose a family of glycosylphosphatidylinositol (GPI)-anchored heparan sulfate proteoglycans that play a role in the control of cell division and growth regulation. So far, six members (GPC1-6) of this family are known in vertebrates. The rat glypican gene 3 (Gpc3) was previously assigned to chromosome Xq36 (Shen et al., 1997). Using standard and radiation cell hybrids, we localized the five other rat glypican genes. PMID- 11474186 TI - Molecular characterization and chromosome assignment of the porcine gene for leukemia inhibitory factor LIF. AB - Leukemia inhibitory factor (LIF) is a pleiotropic cytokine involved in early conceptus development in pig. We isolated a PAC clone containing the porcine LIF gene and determined the complete DNA sequence of the gene, which spans about 6.3 kb and consists of five exons including three alternative first exons (1D, 1M, 1T) spliced onto common second and third exons. The LIF-D transcript encodes a protein of 202 amino acids sharing 87, 84, and 78% identity with respectively human, ovine, and murine leukemia inhibitory factors. The LIF-M and LIF-T transcripts both encode a truncated protein of 158 amino acids. Two SNP markers within untranslated regions of the LIF cDNA were identified. One SNP is located in the 5'-UTR of the alternative exon 1T while the other SNP is located in the 3' UTR of exon 3. Based on fluorescence in situ hybridization and radiation hybrid mapping, the porcine LIF gene was assigned to chromosome 14q2.1-->q2.2. PMID- 11474187 TI - Mouse Sox8 is located between, not within,the t-complex deletions t(w18) and t(h20) on chromosome 17. AB - The SOX family of developmental transcription factors is known to play critical roles in cell lineage specification, fate determination and differentiation during development in diverse phyla. Their importance is underscored by their involvement in a number of human diseases and mouse mutants, and by targeted mutation in mice. SOX8 is broadly expressed during development and is located on human chromosome 16p and within the t-complex on mouse chromosome 17, in the vicinity of two mutations t(w18) and t(h20). Here we analyse mutant genomic DNA to show that the Sox8 gene locus lies outside the deletion regions of both t(w18) and t(h20) and between these deletions. These data exclude Sox8 from contributing to the t(w18) and t(h20) phenotypes, and provide an additional marker for structural characterization of this complex genomic region. PMID- 11474188 TI - Assignment of T cell receptor (TCR) alpha-chain gene (A), beta-chain gene (B), gamma-chain gene (G), and delta-chain gene (D) loci on swine chromosomes by in situ hybridization and radiation hybrid mapping. AB - Using the cDNA sequence of porcine T-cell receptor (TCR) alpha-, beta-, gamma-, and delta-chain genes, we screened a porcine BAC library to isolate clones containing these genes. The isolated BAC clones were confirmed to carry these TCR genes by partial nucleotide sequencing. Among the clones obtained in the present screening, two clones carried both the TCR alpha-chain gene (TRA) and the TCR delta-chain gene (TRD) while one clone each carried only the sequence of either TRA or TRD. This observation demonstrated that TRA and TRD are localized in close proximity on a swine chromosome. Also two clones contained the sequence of the TCR beta-chain gene (TRB), and two clones contained the sequence of TCR gamma chain gene (TRG). Fluorescence in situ hybridization using the above BAC clones as probes revealed that TRA and TRD, TRB, and TRG loci reside on swine chromosomes 7q15.3-->q21, 18q11.3-->q12, and 9q21-->q22, respectively. The chromosome positions of TRA and TRB are consistent with those determined by somatic cell hybrid analysis (Rettenberger et al., 1996). In addition, RH mapping of these genes was performed using the INRA-University of Minnesota RH panel DNA. The result confirmed the position of TRA and TRB reported earlier (http://imprh.toulouse.inra.fr/), and further demonstrated that TRG was located 11 cR away from genetic marker SW989 toward the marker S0019. PMID- 11474189 TI - Organization and chromosomal localization of the human and mouse genes coding for LanC-like protein 1 (LANCL1). AB - We have determined the organization and chromosome location of the human LANCL1 and mouse Lancl1 genes encoding LANCL1, the lanthionine synthetase component C (LanC)-like protein 1. LANCL1 is related to the bacterial LanC family which is involved in the biosynthesis of antimicrobial peptides. The human and mouse genes span 45 kb and 38 kb, respectively, each comprising ten exons. Within the potential promoter regions, several consensus sequences for ubiquitous and tissue specific transcription factors are present, reflecting the expression data. The nucleotide sequence of the previously unknown mouse full-length transcript is also reported here. Fluorescence in situ hybridization analyses assigned the LANCL1 gene to human chromosome 2q34 and the Lancl1 gene to mouse chromosome 1, region C2-C5, in accordance with the known homology. PMID- 11474190 TI - Characterization and chromosomal localization of five canine ATOX1 pseudogenes. AB - We have isolated six ATOX1 loci from the canine genome in BAC clones. Sequence analysis showed that five of these clones correspond to processed pseudogenes. Fluorescent in situ hybridization allowed us to map the genuine ATOX1 gene to CFA4q24-->q31 and the ATOX1 pseudogenes to CFA19q13.1, CFA4q24-->q31, CFA18q24- >q25, CFA9q22.1 -->q22.2 and CFA20q11-->q12. PMID- 11474191 TI - Sequence polymorphisms, allelic expression status and chromosome locations of the chicken IGF2 and MPR1 genes. AB - By screening 26 chicken breeds and lines, DNA polymorphisms were identified in the IGF2 and MPR1 genes, of which mammalian homologues are parentally imprinted, and the GAPD gene, a housekeeping control. Using the polymorphisms as genetic markers, we found that all three genes are expressed biallelically in embryonic tissues. IGF2 and MPR1 were mapped on chicken chromosomes 5 and 3, respectively, by fluorescence in situ hybridization, demonstrating conserved linkage homology between mammals and birds. PMID- 11474192 TI - DNA amplification associated with double minutes originating from chromosome 19 in mouse hepatocellular carcinoma. AB - DNA amplification is associated with genomic instability, the main characteristic of cancer cells, and it frequently involves protooncogenes. Double minute chromosomes (DM) and homogeneously stained regions (HSR) are cytological manifestations of DNA amplification. Gain of chromosome 19 is a recurrent alteration in mouse hepatocellular carcinoma (HCC). In one tumor cell line established from HCC developed in myc transgenic mice, DM derived from chromosome 19 were identified by spectral karyotyping and confirmed by fluorescence in situ hybridization (FISH). A probe generated by PCR from microdissected DM was localized by FISH on normal and HCC-derived cell lines on DM and chromosome 19 at two sites separated by several medium size G-bands. This organization of DM containing amplified sequences from separate loci of the same chromosome, indicates a complex mechanism of DNA amplification, possibly involving more than one gene. DM or HSR were not previously identified in mouse HCC and adult human HCC. The recognition of these loci could lead to the cloning of new genes or identification of known genes important in development or progression of HCC. PMID- 11474193 TI - Genomic organization, chromosome location, and expression analysis of mouse beta synuclein, a candidate for involvement in neurodegeneration. AB - The synuclein family of proteins is a group of primarily brain-expressed polypeptides that show a high degree of amino acid conservation. alpha-Synuclein is the best known of the synuclein family, as it is a major component of the Lewy body, a cytoplasmic inclusion characteristic of Parkinson's disease as well as a variety of related neurodegenerative disorders. With the discovery that mutations in alpha-synuclein can cause Parkinson's disease, a potential role for the other synuclein family members in neurodegenerative disease is being considered. beta Synuclein in particular may deserve special attention, as it is co-expressed with alpha-synuclein at presynaptic nerve terminals, is subject to phosphorylation by Ca(2+) calmodulin protein kinase II, appears important for neural plasticity, and forms aggregates in the brains of patients with Parkinson's disease and a related disorder. To facilitate study of beta-synuclein, we have cloned the mouse beta synuclein gene (Sncb) and determined its genomic organization, size, and intron exon structure. Using an interspecific backcross mapping panel from The Jackson Laboratory, we were then able to localize Sncb to chromosome 13 at the MGD 35.0 cM position. Like the human beta-synuclein gene, Sncb appears to consist of six exons separated by five introns. Unlike the human beta-synuclein gene, the mouse ortholog possesses a variant GC 5' splice donor sequence at the exon 4 - intron 4 boundary in a highly conserved splice junction consensus. Northern blot analysis and Western blot analysis both indicate that Sncb is highly expressed in the brain. Knowledge of the genomic organization and expression pattern of Sncb will allow functional studies of its potential role in neurodegeneration to commence in the mouse. PMID- 11474194 TI - Twelve loci from HSA10, HSA11 and HSA20 were comparatively FISH-mapped on river buffalo and sheep chromosomes. AB - Ten type I loci from HSA10 (IL2RA and VIM), HSA11 (HBB and FSHB) and HSA20 (THBD, AVP/OXT, GNAS1, HCK and TOP1) and two domestic cattle type II loci (CSSM30 and BL42) were FISH mapped to R-banded river buffalo (BBU) and sheep (OAR) chromosomes. IL2RA (HSA10) maps on BBU14q13 and OAR13q13, VIM (HSA10) maps on BBU14q15 and OAR13q15, HBB (HSA11) maps on BBU16q25 and OAR15q23, FSHB (HSA11) maps on BBU16q28 and OAR15q26, THBD (HSA20) maps on BBU14q15 and OAR13q15 while AVP/OXT, GNAS1, HCK, and TOP1 (HSA20) as well as CSSM30 and BL42 map on the same large band of BBU14q22 and OAR13q22. All loci were mapped on the same homologous chromosomes and chromosome bands of the two species, and these results agree with those earlier reported in cattle homologous chromosomes 15 and 13, respectively, confirming the high degree of both banding and physical map similarities among the bovid species. Indirect comparisons between physical maps achieved on bovid chromosomes and those reported on HSA10, HSA11 and HSA20 were performed. PMID- 11474195 TI - Chromosomal distribution of the telomere sequence (TTAGGG)(n) in the Equidae. AB - Telomeres are a class of repetitive DNA sequences that are located at chromosome termini and that act to stabilize the chromosome ends. The rapid karyotypic evolution of the genus Equus has given rise to ten taxa, all with different diploid chromosome numbers. Using fluorescence in situ hybridization (FISH) we localized the mammalian telomere sequence, (TTAGGG)(n), to the chromosomes of nine equid taxa. TTAGGG signal was located at chromosome termini in all species, however additional signal was seen at interstitial sites on some chromosomes in the Burchell's zebra, Equus quagga burchelli, the Hartmann's zebra, Equus zebra hartmannae, and at large heterochromatin-associated regions on the chromosomes of the donkey, Equus asinus. The interstitial signal in the zebras may be a relic of an ancient telomere-telomere fusion and mark the point at which two ancestral chromosomes may have fused. For the donkey, the heterochromatin-associated signal may represent degenerate telomere-like satellite sequences and identify a second type of satellite DNA for this taxon. PMID- 11474196 TI - The human FGF gene family: chromosome location and phylogenetic analysis. PMID- 11474197 TI - Assignment of SH3KBP1 to human chromosome band Xp22.1-->p21.3 by in situ hybridization. PMID- 11474198 TI - Assignment of the human histone deacetylase 6 gene (HDAC6) to X chromosome p11.23 by in situ hybridization. PMID- 11474199 TI - Assignment of the human histone deacetylase 4 gene (HDAC4) to chromosome 2q37.2 by in situ hybridization. PMID- 11474200 TI - Assignment of the human metastasis-associated gene 1 (MTA1) to human chromosome band 14q32.3 by fluorescence in situ hybridization. PMID- 11474201 TI - Assignment of the fatty acid synthase (FASN) gene to bovine chromosome 19 (19q22) by in situ hybridization and confirmation by somatic cell hybrid mapping. PMID- 11474202 TI - Assignment of an ionotropic glutamate receptor-like gene (GRINL1A) to human chromosome 15q22.1 by in situ hybridization. PMID- 11474203 TI - Assignment of the homeobox A10 gene (HOXA10) to porcine chromosome SSC18q23-->q24 by FISH and confirmation by hybrid panel analyses. PMID- 11474204 TI - Assignment of human NADH dehydrogenase (ubiquinone) 1 beta subcomplex 3 (NDUFB3) and of its four pseudogenes to human chromosomes 2q31.3, 1p13.3-->p13.1, 9q32- >q34.1, 14q22.3-->q23.1 and 14q32.2 by radiation hybrid mapping. PMID- 11474205 TI - Assignment of myotrophin to human chromosome band 7q33-->q35 by in situ hybridization. PMID- 11474206 TI - Consanguinity and prereproductive mortality in the Utah Mormon population. AB - To test the effects of parental consanguinity on mortality among offspring, inbreeding coefficients were estimated for 303,675 members of the Utah Mormon population who were born between 1847 and 1945. Although consanguinity has been relatively rare in this population, the large sample size permitted the identification of more than 3,500 inbred offspring. Among the offspring of unrelated parents, 13.2% died before the age of 16. Significant elevations in prereproductive mortality were seen among the offspring of first-cousin marriages (22%) and among the offspring of closer unions (32%). The cor- responding relative risks are 1.70 (95% confidence limits = 1.52, 1.91) and 2.41 (95% confidence limits = 1.59, 3.41), respectively. Other categories of relationship did not produce significant elevations in offspring mortality. Similar results were obtained when a case-control approach was used to remove the effects of socioeconomic variation. Consistent with many other studies of populations with low consanguinity rates, this population experienced a relatively high absolute increase in mortality among the offspring of first-cousin marriages (9%). Preliminary evidence is offered for the hypothesis that mortality differentials are larger in populations with low inbreeding and low mortality because nongenetic causes of death do not obscure the effects of consanguinity. PMID- 11474207 TI - Association tests in nuclear families. AB - We present a conditional likelihood approach for testing linkage disequilibrium in nuclear families having multiple affected offspring. The likelihood, conditioned on the identity-by-descent (IBD) structure of the sibling genotypes, is unaffected by familial correlation in disease status that arises from linkage between a marker locus and the unobserved trait locus. Two such conditional likelihoods are compared: one that conditions on IBD and phase of the transmitted alleles and a second which conditions only on IBD of the transmitted alleles. Under the log-additive model, the first likelihood is equivalent to the allele counting methods proposed in the literature. The second likelihood is valid under the added assumption of equal male and female recombination fractions. In a simulation study, we demonstrated that in sibships having two or three affected siblings the score test from each likelihood had the correct test size for testing disequilibrium. They also led to equivalent power to detect linkage disequilibrium at the 5% significance level. PMID- 11474208 TI - Localization of a recessive juvenile cataract mutation to proximal chromosome 7 in mice. AB - OBJECTIVE: To localize the chromosomal position of a novel cataract mutation (juvenile recessive cataract; jrc) in mice. METHODS: A mapping population was developed by crossing cataract males (albino MH) to wild-type females (black C57BL/6J). F1 females were backcrossed to albino MH males with cataracts. RESULTS: The results were consistent with a model of a single autosomal recessive gene [153 cataract, 169 wild-type; chi2 = 0.8, 1 degree of freedom (d.f.), p > 0.35]. Linkage with the albino (tyrosinase; Tyr) locus was evident (chi2 = 61.5, 1 d.f., p < 0.0001), implicating chromosome 7 as the location of jrc. Recombination percentages (+/- SE) between jrc and D7Mit340 (1.2 cM location), D7Mit227 (16.0 cM) and D7Mit270 (18.0 cM) were 17.1 +/- 2.1, 3.7 +/- 1.1 and 6.2 +/- 1.3%, respectively. Multi-point mapping determined that the most likely order of these loci is D7Mit340 - jrc - D7Mit227 - D7Mit270 - Tyr. Although animals with the mutant phenotype appeared to have little or no sense of sight, their growth was not different (p >0.20) from that of normal mice. CONCLUSION: The jrc mutation model may be useful in the study of the genetics of cataracts in other animal species, including humans. PMID- 11474209 TI - The score statistic of the LD-lod analysis: detecting linkage adaptive to linkage disequilibrium. AB - We study the properties of a modified lod score method for testing linkage that incorporates linkage disequilibrium (LD-lod). By examination of its score statistic, we show that the LD-lod score method adaptively combines two sources of information: (a) the IBD sharing score which is informative for linkage regardless of the existence of LD and (b) the contrast between allele-specific IBD sharing scores which is informative for linkage only in the presence of LD. We also consider the connection between the LD-lod score method and the transmission-disequilibrium test (TDT) for triad data and the mean test for affected sib pair (ASP) data. We show that, for triad data, the recessive LD-lod test is asymptotically equivalent to the TDT; and for ASP data, it is an adaptive combination of the TDT and the ASP mean test. We demonstrate that the LD-lod score method has relatively good statistical efficiency in comparison with the ASP mean test and the TDT for a broad range of LD and the genetic models considered in this report. Therefore, the LD-lod score method is an interesting approach for detecting linkage when the extent of LD is unknown, such as in a genome-wide screen with a dense set of genetic markers. PMID- 11474210 TI - Novel mutations (H337R and 238-362del) in the CPS1 gene cause carbamoyl phosphate synthetase I deficiency. PMID- 11474211 TI - Statistical considerations for genome-wide scans: design and application of a novel software package POLYMORPHISM. AB - OBJECTIVE: Given the cost and complexity of genome-wide scans, optimization of study design is of critical importance. Available algorithms only partially satisfy this need. We designed a software package called 'POLYMORPHISM' to meet these needs. METHODS: The program is designed to calculate linkage parameters for both 'single-point' and 'two-point' settings that are applicable also to incompletely informative microsatellite markers. In single-point analysis, the heterozygosity, polymorphism information content (PIC) and linkage information content (LIC) statistics based on marker allele frequencies are provided. In two point analysis, joint PIC values for two markers, the conditional probability of detecting linkage phase, the frequency of double heterozygotes and the expected number of informative meioses are calculated. RESULTS: Results were obtained using S.A.G.E./DESPAIR (Design of Linkage Studies Based on Pairs of Relatives) in addition to applying this program to a Centre d'Etude du Polymorphisme pedigree derived genotyping data set, which estimated critical parameters used in a two stage genome scan. A single nucleotide polymorphism (SNP)-based one-stage genomic screen strategy is also considered. CONCLUSIONS: LIC values are crucial for getting accurate estimates on those parameters that are important for a two-stage genome screening study. Optimization of the cost-effectiveness of an SNP-based genomic screen strategy is possible by modeling a balance between marker information content and marker density. PMID- 11474212 TI - Haemochromatosis gene mutations in Finns, Swedes and Swedish Saamis. AB - Frequencies of three different mutant haemochromatosis (HFE) alleles (282Tyr, 63Asp and 65Cys) were studied in three northern European populations, i.e. Finns, Swedes and Swedish Saamis. In Finns and Swedes the allele frequencies were within the range found in other populations from northern and western Europe. The Saamis differed from the Swedes with respect to all mutant alleles. Lower frequencies compared to Swedes were found for the 282Tyr (p = 0.0046) and 63Asp (p = 0.034) alleles, whereas the frequency of the 65Cys allele was higher (p = 0.046) in the Saamis. The total distribution of HFE alleles in Saamis showed a highly significant difference from that in Swedes (chi2 = 16.7, 3 d.f., p = 0.0008). These results further underline the genetic uniqueness of the Saamis. PMID- 11474213 TI - Improved power of sib-pair linkage analysis using measures of complex trait dynamics. AB - The influence of epistasis on a quantitative trait can reduce the power of linkage analysis to identify the underlying loci. In the present study, we simulated a complex trait derived from a dynamic one-locus gene expression system with epistasis arising from feedback regulation and tested the power of sib-pair linkage analysis methods for detecting the underlying quantitative trait locus (QTL). Using this simple genetic architecture, we demonstrate that the power of sib-pair linkage analysis can be greatly improved if measures of complex trait dynamics are considered. PMID- 11474214 TI - The endocrine disrupters butyl benzyl phthalate and bisphenol A increase the expression of progesterone receptor messenger ribonucleic acid in the preoptic area of adult ovariectomized rats. AB - Butyl benzyl phthalate (BBP) and bisphenol A (BPA), so-called endocrine disrupters, are known to mimic the action of estrogens: they are thus liable to influence reproductive functions. Since little is known about their action on gene expression in the adult hypothalamus, we examined the effects of these chemicals on the expression of estrogen-regulated mRNAs, i.e., progesterone receptor (PR) mRNA, preproenkephalin (PPE) mRNA and neurotensin (NT) mRNA, in the hypothalamus and pituitary of adult female rats. Two weeks after ovariectomy, rats were subcutaneously injected with 10 mg BBP, 10 mg BPA, or 10 microg 17 beta estradiol (E(2)) in sesame oil, or with sesame oil alone as a control. Twenty four hours after the injection, tissues including the preoptic area (POA), mediobasal hypothalamus (MBH) and anterior pituitary were collected. Northern blot revealed that injection of E(2) resulted in expected changes, i.e., significant increases in PR mRNA in the POA, MBH and anterior pituitary, and in PPE mRNA in the MBH. We also found that injection of BPA significantly increased PR mRNA in the POA and anterior pituitary, while injection of BBP increased PR mRNA in the POA and anterior pituitary, although the increase in the anterior pituitary was not significant. No significant effect of E(2), BPA, or BBP on NT mRNA in the POA was detected. The present study demonstrates that the two endocrine disrupters BPA and BBP can increase the expression of PR mRNA in the POA of adult ovariectomized rats. PMID- 11474215 TI - Nitric oxide mediates the inhibitory effect of tumor necrosis factor-alpha on prolactin release. AB - Tumor necrosis factor-alpha (TNF-alpha) is a pleiotropic cytokine that markedly affects neuroendocrine functions. This cytokine is expressed in the anterior pituitary where its receptors are also present. Nitric oxide (NO) is synthesized in gonadotropes and folliculo-stellate cells of the anterior pituitary. Since NO directly inhibits prolactin secretion, we investigated the involvement of NO in the inhibitory effect of TNF-alpha on prolactin release from anterior pituitary cells of female rats. The presence of L-NAME (1 mM), an inhibitor of NO synthase (NOS), in the incubation medium significantly blunted the inhibition of prolactin release produced by TNF-alpha (50 ng/ml). TNF-alpha increased nitrite release to the incubation medium. The activity of NOS as measured by [(14)C]citrulline production was significantly enhanced when anterior pituitary cells were incubated with TNF-alpha for 8 h or more. Also, TNF-alpha induced iNOS gene expression in anterior pituitary cells as assessed by reverse transcriptase polymerase chain reaction. The current results indicate that NO is involved in the inhibitory effect of TNF-alpha on prolactin secretion and that TNF-alpha induces iNOS transcription and stimulates NO synthesis in anterior pituitary cells. PMID- 11474216 TI - Effects of loperamide and other opioid-related substances on the transcriptional regulation of the rat pro-opiomelanocortin gene in AtT20 cells. AB - Although opioid peptides are involved in the regulation of the hypothalamic pituitary-adrenal axis, their role in pro-opiomelanocortin (POMC) gene expression at the pituitary level is not known. We therefore examined the effects of opioid receptor agonists, including recently discovered endogenous opioid peptides, on POMC gene expression using the AtT20PL cell line, a subclone of AtT20 in which the rat POMC 5'-promoter-luciferase fusion gene was stably incorporated. The endogenous mu-opioid receptor agonists endomorphin 1 and 2 had no effect on either basal or corticotropin-stimulating-hormone-induced POMC expression. This was also the case with the delta-agonist BUBUC, the kappa-agonist U50488H and the orphan receptor agonist orphanin FQ. In contrast, the synthetic mu-agonist loperamide significantly inhibited basal and yet enhanced cAMP-induced POMC expression. The inhibitory effect of loperamide was mimicked by the calmodulin antagonist W7 and antagonized by the calcium channel blocker nifedipine, whereas neither the inhibitory nor the enhancing effect of loperamide was influenced by the opioid antagonist naloxone. These results suggest that the synthetic mu agonist loperamide has a modulatory effect on the 5'-promoter activity of the POMC gene. This effect does not seem to be mediated through the classical mu opioid receptor but rather in part through a calcium/calmodulin-related mechanism. PMID- 11474217 TI - Vascular endothelial growth factor production and regulation in rodent and human pituitary tumor cells in vitro. AB - Angiogenesis, the formation of a new blood supply, is an essential step in tumorigenesis. Although vascular endothelial growth factor (VEGF) is known to be a very potent angiogenic factor in most solid tumors, little is known about its production and regulation in pituitary adenomas. We have investigated basal and stimulated VEGF production by rodent pituitary tumor cells (mouse corticotrope AtT20, rat lactosomatotrope GH3, mouse gonadotrope alpha T3-1 and mouse folliculostellate TtT/GF cells), and by hormone-inactive (27), corticotrope (9), lactotrope (3) and somatotrope (21) human pituitary adenoma cell cultures. All 4 pituitary cell lines secreted VEGF, which in the case of AtT20, GH3 and TtT/GF cells was inhibited by approximately 50% by dexamethasone. TtT/GF cells were the most responsive to the different stimuli used since basal values were augmented by pituitary adenylate cyclase activating polypeptide-38 (PACAP-38), interleukin 6 (IL-6), transforming growth factor-alpha (TGF-alpha), IGF-I and the somatostatin analogue ocreotide. However, in GH3, AtT20 and alpha T3-1 cells, basal VEGF levels where not enhanced with any of the stimuli tested. The majority of the human adenomas tested (92%) basally secreted measurable VEGF which was inhibited by dexamethasone in most cases (84%). VEGF levels were increased in hormone inactive adenomas, somatotrope tumors and prolactinomas by TGF-alpha, PACAP-38, and 17 beta-estradiol, respectively. In conclusion, pituitary tumor cells are capable of producing VEGF which may be involved in tumoral angiogenesis. Our results concerning the suppression of VEGF by dexamethasone suggest that glucocorticoids may have anti-angiogenic properties and therefore therapeutic relevance for the treatment of pituitary adenomas. PMID- 11474218 TI - Hypothalamic circuitry of neuropeptide Y regulation of neuroendocrine function and food intake via the Y5 receptor subtype. AB - Neuropeptide Y (NPY) displays diverse modes of action in the CNS including the modulation of feeding behavior, gonadotropin releasing hormone release, and stress responses. Many of the above physiological actions have been at least partially attributed to actions of NPY on the NPY Y5 receptor subtype. We utilized an antibody directed against the NPY Y5 receptor to characterize the distribution of this receptor in the rat brain. Using Western blot analysis, this antibody recognized a single major band at approximately 57 kD. To further verify the specificity of the antibody, animals were treated for 5 days with antisense oligonucleotides for the Y5 receptor. The antisense treatment significantly reduced food intake and body weight. Furthermore, the Y5 antibody detected a significant decrease in Y5 receptor protein. Y5-like immunoreactivity (-ir) was observed throughout the hypothalamus, thalamus, hippocampus and cortex. Double label immunofluorescence demonstrated that Y5-ir was colocalized with the following neuronal phenotypes in the hypothalamus, gonadotropin-releasing hormone, neurophysins, corticotropin-releasing hormone, and gamma-amino butyric acid. In addition, functional interactions were demonstrated by the presence of close appositions of NPY fibers with Y5-ir expressing cells. The wide distribution of the Y5 receptor-ir, as well as the colocalization within specific neuronal populations, agrees with the distribution of the Y5 receptor mRNA and the known physiological roles of the NPY/Y5 system. The role of the NPY/Y5 receptor system as a mediator between signals of peripheral energy availability and reproductive neuroendocrine function is discussed. PMID- 11474219 TI - Lack of decrease in hypothalamic and hippocampal glucocorticoid receptor mRNA during starvation. AB - We have shown in a previous study that high corticosterone levels during repeated immobilization stress result in a reduction of glucocorticoid receptor (GR) mRNA in the hypothalamic paraventricular nucleus (PVN) and the hippocampus. The reduction of GR presumably accounts for loss of or decrease in glucocorticoid negative feedback, and thus hyperfunction of the hypothalamic-pituitary adrenocortical (HPA) axis persists during chronic stress. Starvation is a stress state in which the counterregulatory responses against the loss of food occur in the central nervous system. We explored the impact of starvation on the HPA axis, GR and mineralocorticoid receptor (MR) mRNAs in the hippocampus, the PVN, and the anterior pituitary (AP) of rats. Rats were starved for 4 days and sacrificed in the morning. Starved rats showed high levels of plasma corticosterone, whereas neither plasma corticotropin (ACTH), AP proopiomelanocortin (POMC) mRNA nor AP type-1 corticotropin-releasing hormone (CRH) receptor mRNA was altered in the starved rats. In the presence of high corticosterone, starvation resulted in a decrease in both CRH mRNA and type-1 CRH receptor mRNA in the PVN. Consistently, the starved rats did not show any changes in GR mRNA in the hippocampus (CA1-2, CA3, and dentate gyrus), the PVN or the AP despite the elevation of plasma corticosterone. A significant decrease in MR mRNA was seen in the dentate gyrus and the AP, but not in CA1-2, CA3 or PVN. The lack of reduction of GR may be one of the organism's counterregulatory responses during starvation, which allows an intact glucocorticoid negative feedback, thereby resulting in decreased anorectic neuropeptide levels, namely CRH, in the PVN. The results also indicate that GR mRNA in the hippocampus and other brain regions is not solely regulated by circulating glucocorticoids. The mechanism underlying the regulation of GR mRNA in the central nervous system remains to be clarified. PMID- 11474220 TI - Central stimulatory influence of oxytocin on preovulatory gonadotropin-releasing hormone requires more than the median eminence. AB - The study was designed to determine whether the ability of central oxytocin (OT) to stimulate gonadotropin-releasing hormone (GnRH) on the afternoon of proestrus (PE) in the cycling female rat is mediated at the level of GnRH terminals within the median eminence (ME), or at higher hypothalamic levels where GnRH cell bodies and axons are located. Determining the location of this OT effect in vivo has proven difficult. Therefore, an in vitro system utilizing ME or basal hypothalamic (BH) explants containing GnRH terminals, or GnRH neurons including the cell bodies, axons and terminals, respectively, were harvested from regular cycling female rats at 15:00 h on PE or diestrus (DI). The explants were allowed to preincubate in Krebs Ringer Bicarbonate Buffer containing glucose, ascorbic acid, calcium, and a metalloprotease inhibitor (KRBG) and enriched with 95% O(2)/5% CO(2) at 37 degrees C until a stable baseline release of GnRH was achieved (30 min). The 0.05 level of probability was used as the minimum criterion of significance in all experiments. The ability of OT (10(-15)--10(-9) M) to stimulate the release of GnRH was determined in both ME and BH explants on PE and DI. The results demonstrated a sensitive, dose-dependent ability of OT to stimulate GnRH release from PE BH explants which was observed only in PE. Furthermore, OT failed to significantly stimulate GnRH release from ME explants on either PE or DI. The data indicate that the PE BH explant paradigm can be used to examine the manner and mechanisms by which OT influences GnRH release on the afternoon of PE. Furthermore, the results indicate for the first time that the stimulatory action of OT by itself on preovulatory GnRH release in cycling female rats is not mediated at the level of the GnRH terminals within the ME, but requires neuronal interactions and mechanisms within the BH explants. PMID- 11474221 TI - Serotonin 5-HT(1A) receptor mRNA expression in dorsal hippocampus and raphe nuclei after gonadal hormone manipulation in female rats. AB - Female ovarian steroids influence mood and cognition, an effect presumably mediated by the serotonergic system. A key receptor in this interplay may be the 5-HT(1A) receptor subtype. We gave adult ovariectomized female rats subcutaneous pellets containing different dosages of 17 beta-estradiol alone or in combination with progesterone, or placebo pellets, for 2 weeks. 5-HT(1A) receptor mRNA levels were analyzed by in situ hybridization in the dorsal hippocampus, dorsal and median raphe nuclei, and entorhinal cortex. Estradiol treatment alone reduced 5 HT(1A) gene expression in the dentate gyrus and the CA2 region (17 and 19% decrease, respectively). Estradiol combined with progesterone supplementation increased 5-HT(1A) gene expression versus placebo in the CA1 and CA2 subregions of the dorsal hippocampus (16 and 30% increase, respectively). Concomitantly, 5 HT(1A) mRNA expression was decreased by 13% in the ventrolateral part of the dorsal raphe nuclei, while no changes were found in the median raphe nucleus and entorhinal cortex. Chronic effects of ovarian hormones on 5-HT(1A) receptor mRNA expression appear tissue-specific and involve hippocampal subregions and the raphe nuclei. Modulation of 5-HT(1A) receptor gene expression may be of importance for gonadal steroid effects on mood and cognition. PMID- 11474222 TI - New vistas in anti-inflammatory therapy. PMID- 11474223 TI - Dialysis in non-renal organ (liver) transplantation. AB - The renal management of acute hepatic failure and liver transplantation requires an understanding of the features of liver failure and the causes of liver graft dysfunction. The management of any underlying pathology in addition to supportive care is fundamental to successful management and a return to independent renal function. These issues are discussed particularly in relationship to a case history involving a patient presenting with acute fulminant liver failure secondary to a paracetamol overdose who was successfully treated by liver transplantation and continuous veno-venous haemodiafiltration. The liver can be successfully transplanted but acute renal failure is a severe complication post transplantation. Its appearance can be predicted in patients with pre-transplant renal dysfunction, severe graft dysfunction, or both. It may be avoided through careful selection of transplant recipients and correct timing of transplantation. Prevention of renal failure, appropriate patient selection for transplantation and timely procurement of a donor organ are vital for best use of limited donor resources. Treatment success depends on patient and donor selection, skilled surgeons, careful post-operative care, and successful immunosuppression. PMID- 11474224 TI - Urinary copper excretion in type 2 diabetic patients with nephropathy. AB - BACKGROUND/AIMS: The aim of this study was to examine the relationship between the degree of urinary copper excretion and stages of diabetic nephropathy. METHODS: Copper, ceruloplasmin and albumin concentrations were measured in serum and urine samples from 41 type 2 diabetic outpatients with different stages of nephropathy and from 10 healthy controls. The copper/albumin and copper/ceruloplasmin ratios in serum and urine were determined. Furthermore, we examined whether free copper ions are dissociated from ceruloplasmin under various pH conditions. RESULTS: Urinary copper concentrations significantly increased only in macroalbuminuric patients. The copper/ceruloplasmin and copper/albumin ratios in urine were consistently greater than those in serum which were not different between patients and healthy controls except the copper/albumin ratio in macroalbuminuric patients. The ratios in urine decreased in parallel with the progression of nephropathy. Copper was found to be released from ceruloplasmin under acidic conditions. CONCLUSION: Urinary copper excretion in healthy controls may be the result of dissociation from the albumin-copper complex of serum during its passage through the kidney. In diabetic patients with advanced nephropathy, urinary copper excretion may be due to dissociations from both copper-albumin and ceruloplasmin-copper complexes filtered through the damaged glomerulus. Overloading of urinary copper to damaged renal tubules may play some roles in the progression of nephropathy in patients with advanced nephropathy. PMID- 11474225 TI - Significance of ACE genotypes and medical treatments in childhood focal glomerulosclerosis. AB - BACKGROUND: There is little information on the significance of angiotensin converting enzyme (ACE) genotypes and medical treatments in children with primary focal segmental glomerulosclerosis (FSGS). METHODS: A multicenter retrospective study was performed on the role of ACE genotypes and medical treatments in 43 Japanese children with FSGS (20 males and 23 females), including 17 children who progressed to end-stage renal failure during the mean observation period of 6.9 +/- (SD) 5.0 years. RESULTS: The incidence of the D allele of the ACE gene was higher in the whole group of 43 children with FSGS and in a subgroup of 28 steroid-resistant FSGS children (p < 0.05) than in the 130 children of the healthy control group (0.48, 0.48, and 0.33, respectively). ACE genotypes did not affect renal survival in the whole FSGS group nor in the steroid-resistant subgroup. Among the 28 steroid-resistant children, treatment with ciclosporin was effective in delaying the development of end-stage renal failure (p = 0.044), independently of other treatment regimens. CONCLUSION: The present study of Japanese children with FSGS showed that the D allele of the ACE gene is associated with the development of FSGS, but not associated with the progression of FSGS which was greatly ameliorated with ciclosporin, irrespective of ACE genotypes. PMID- 11474226 TI - Study on concretions developed around urinary catheters and mechanisms of renal calculi development. AB - AIMS: To study the structure and composition of encrustation and concretions developed on urinary catheters to better understand their formation mechanism to be able to prevent them. METHODS: The surface of catheters was studied by direct and scanning electron microscopy observation. In vitro formation of encrustations was performed in synthetic urine. RESULTS: The surface of catheters was covered by a continuous layer of organic matter, on which a thin scale consisting of crystals of calcium oxalate monohydrate (COM), uric acid anhydrous or calcium phosphate developed. Encrustations observed on catheters generally exhibited the same composition as the previously developed renal calculi. In catheters collected from patients without previous episodes of renal calculi or with previous episodes of infected renal calculi in which infection was afterwards eradicated, on the first organic layer, in that case plate-like COM crystals forming a columnar layer were observed. In vitro experiments demonstrated that COM columnar structures were only formed when normocalciuric urine containing organic matter was used, and the presence of crystallization inhibitors, as phytate, notably delayed their formation. CONCLUSION: Calcium oxalate was the main crystalline phase developed on catheters inserted in patients, specially in the absence of urinary infection or urinary pH values <5.5 and high urinary uric acid levels. Thus, prophylaxis of encrustations may consist of preventive measures usually applied in cases of recurrent idiopathic calcium oxalate urolithiasis. PMID- 11474227 TI - Combined treatment with low-dose pravastatin and fish oil in post-renal transplantation dislipidemia. AB - BACKGROUND: The most common cause of post-transplant dyslipidemia is the use of corticosteroids and cyclosporin-A (CyA). The HMG-CoA reductase inhibitors have emerged as the agents of first choice in the treatment of post-transplant hyperlipidemia in combination with low fat diet. The objective of this study was to evaluate the efficacy of combined treatment with low-dose pravastatin and fish oil in post-renal transplantation dislipidemia. PATIENTS AND METHODS: Twenty-four renal transplant patients, 15 men and 9 women aged from 30 to 60 years with stable renal function were included in this study. All patients were transplanted from living related donors and were given a stable triple immunosuppressive therapy, with methylprednisolone, azathioprine and CyA. All patients were also given a standard diet containing 1 g/kg BW protein, reducing the daily fat to less than 30%, and maintaining at least a 1:1 ratio of saturated to polyunsaturated (or monounsaturated) fats. A dosage of 20 mg pravastatin (pravachol) and 1 g of fish oil (prolipid) were added to the diet after dinner, according to our protocol. Blood samples were taken after each study period for total cholesterol, LDL-cholesterol, triglycerides, Apo A(1), Apo B, Lp(a), creatinine, CPK and fibrinogen determination. RESULTS: At the end of the therapeutic protocol with pravastatin a significant reduction (p < 0.02) of total and LDL-cholesterol was observed, but no significant change in triglycerides, HDL, Lp(a), Apo A(1), Apo B and fibrinogen was shown. At the end of the therapeutic protocol with pravastatin and fish oil supplement significant changes were seen in TC (p < 0.02), TG (p < 0.03), LDL-C (p < 0.03), Apo A(1) (p < 0.04) and Apo B (p < 0.05) concentrations. There were no significant changes in HDL-C and Lp(a) concentrations. Renal function and cyclosporine levels were not changed during and after the study. CPK was increased only in one case. CONCLUSIONS: It is suggested that if the response to the diet is inadequate, the use of combined treatment with low-dose pravastatin and fish oil is a more effective strategy than the pravastatin treatment alone for changing the lipid profile after renal transplantation. PMID- 11474228 TI - Elevated interleukin-18 levels in the urine of nephrotic patients. AB - BACKGROUND/AIM: The etiology of minimal-change nephritic syndrome (MCNS) is obscure. It has been speculated that T cells play a role in the pathogenesis of MCNS. Interleukin (IL)-18, a novel immunoregulatory cytokine with potent inferon gamma-inducing activities, may play an important role in T-helper type 1-mediated immune responses. To examine further the possible role of IL-18 in nephrotic syndrome (NS), in the present study we measured IL-18 levels in the urine in different clinical stages of MCNS. The aim of this study was to investigate the involvement of IL-18 in MCNS. METHODS: Urine samples were obtained from 20 MCNS patients. The disease controls included 20 patients with IgA nephropathy. The samples were assayed for IL-18 protein by a sandwich enzyme-linked immunosorbent assay. RESULTS: Compared with normal controls, significantly increased urinary levels of IL-18 were detected in MCNS patients with the NS. The urinary IL-18 (uIL-18) levels correlated with the degree of proteinuria in MCNS patients. Moreover, when individual MCNS patients were followed through their clinical illness, uIL-18 levels were increased during the active phase and decreased as the patients went into remission. CONCLUSIONS: These results indicate that uIL- 18 is detectable in a subgroup of patients with active NS and correlates to their disease activity in patients with MCNS. Our findings support the notion that IL 18 may play a role in the pathophysiology of NS. PMID- 11474229 TI - Renal localisation of rat cysteine dioxygenase. AB - BACKGROUND/AIMS: Cysteine dioxygenase (CDO, EC 1.13.11.20) catalyses the conversion of cysteine to cysteine sulphinic acid and controls the rate-limiting step of sulphate production. Many neurological and non-neurological diseases are associated with abnormalities in CDO activity, giving rise to reduced availability of sulphate. The importance of the kidney in the sulphation of xenobiotics has long been recognised, but little is known about the renal expression of key enzymes in this pathway. In order to address this, this report demonstrates the expression of CDO in the kidney. METHODS: Two previously characterised antibodies were used to investigate the localisation and expression of CDO using immunohistochemistry, in-situ hybridisation and Western blotting. RESULTS: Renal CDO was shown to exist as a 68-kDa protein, which was unaffected by levels of cysteine and methionine that had been previously shown to induce hepatic CDO. CDO protein expression was present in the proximal convoluted tubules of the cortex and the collecting ducts of both the medulla and papilla. DISCUSSION: These results suggest that renal CDO is immunologically identical to that of the liver. Its expression in the kidney tubules, the major site of sulphation in the kidney, suggests that CDO in the kidney may play a role in both xenobiotic metabolism and sodium and water homeostasis. PMID- 11474230 TI - Hyaluronan and rat renal fibroblasts: in vitro studies. AB - Hyaluronic acid (HA) is a ubiquitous component of extracellular matrix. After tissue injury, HA appears in greater abundance during the inflammatory response and the phase of clearance of cell and matrix debris, before collagen production and matrix degradation. The aim of this study was to examine whether normal rat renal fibroblasts were capable of HA synthesis and to determine the effect of HA on in vitro collagen production in a series of normal rat cortical fibroblast cultures. Fibroblast cultures from both renal cortex and medulla were established from adult Sprague-Dawley rats. HA synthesis was measured by radioimmunoassay, and incorporation of (3)H-proline into collagen was used to determine collagen synthesis. Fibroblasts were defined on the basis of morphology and alpha smooth muscle actin immunohistochemistry. HA synthesis was measured in both renal cortical and medullary fibroblasts at passage 3 for both 24 and 48 h in 5 animals and expressed as a fraction of protein content. HA was synthesized by both cortical and medullary fibroblasts; however, cortical fibroblasts produced less HA than medullary fibroblasts at both 24 h (p = 0.05) and 48 h (p = 0.02). In normal cortical fibroblasts, exogenous HA suppressed overall total (cell and media) collagen production after a 22-hour labelling period (p = 0.002 compared to controls). Decreased collagen production was also found individually in cell (p = 0.02) and media fractions (p = 0.01). Both cortical and medullary fibroblasts are capable of synthesizing HA in vitro. Furthermore, the findings in this study suggest that HA may be an important mediator in reducing renal cortical fibroblast collagen production and may play an important role in limiting renal interstitial scarring. PMID- 11474231 TI - Effect of methyldopa on renal function in rats with L-NAME-induced hypertension in pregnancy. AB - BACKGROUND: Pregnancy-induced hypertension is characterized by an increased sympathetic activity and probably by a decreased synthesis/activity of nitric oxide. The aim of the present study is to evaluate whether the beneficial action of the sympathetic antagonist methyldopa (a first-choice hypotensive agent in the treatment of PIH) may be associated to changes in nitric-oxide synthesis. METHODS: Forty pregnant Wistar rats received L-NAME (NO synthase inhibitor, 9-10 mg/kg/day) from mid-pregnancy (day 11) through to term. Some rats were treated with daltroban (TxA receptor antagonist, 60 mg/kg/day), diltiazem (calcium channel blocker, 30 mg/kg/day), methyldopa (central adrenergic antagonist, 400 mg/kg/day) or L-arginine (260 mg/kg/day) from mid-pregnancy. The effect of the different treatments on systolic blood pressure (SBP), creatinine clearance (CCR), urine protein excretion (UP) and urinary nitrate excretion (UNO(3), representing urine NO metabolite) were evaluated and the results compared with those found in normal pregnancy. Normal pregnant rats receiving similar treatment were used as controls. RESULTS: In normal pregnant (P) rats, SBP values decreased from 94 +/- 2 to 83 +/- 3 mm Hg at the end of pregnancy (p < 0.01) and CCR augmented significantly. Drug treatment had no significant effect. In NAME treated rats, at the same period, the SBP augmented from 92 +/- 1 to 129 +/- 1.8 mm Hg (p < 0.01). At the end of pregnancy, NAME rats had significantly lower CCR values and higher UP excretion when compared with P rats. UNO(3) increased significantly in P and in P rats treated with methyldopa. As expected, in NAME rats UNO(3) excretion was significantly reduced. Treatment with methyldopa normalized SBP, improved CCR and proteinuria and was associated with an increase in UNO(3). Similar results were obtained with L-arginine treatment. Diltiazem lowered SBP significantly but had no effect on renal function or UNO(3) and daltroban had no effect. CONCLUSION: The increased UNO(3) found in NAME rats treated with methyldopa suggests that the vasoconstriction secondary to chronic NO inhibition may be partially related to an increased sympathetic activity. The efficient action of the sympathetic antagonist methyldopa may be due not only to its antihypertensive effects but also by its stimulating effect on NO synthesis leading also to an improvement of renal function. PMID- 11474232 TI - mRNA expression of the IGF system in the kidney of the hypersomatotropic rat. AB - AIM: To examine the response of the insulinlike growth factor (IGF) system in the kidney during a state of extreme growth. METHODS: We studied the mRNA expression of IGF-I, IGF-I receptor, and IGF-binding proteins (BP) using sensitive RNase protection assays following subcutaneous implantation of growth hormone pituitary cells (GH(3)) in rats. RESULTS: Within 5 weeks, the serum GH levels increased from 18.0 +/- (SE) 5.0 ng/ml in control animals to 389.8 +/- 30.3 ng/ml in GH(3) rats (n = 5, p < 0.001). The circulating IGF-I levels were also elevated. The kidney weights increased from 0.74 +/- 0.01 g in controls to 1.06 +/- 0.03 g in GH(3) animals (n = 5, p < 0.001). Similar changes were observed at week 10. The renal IGF-I mRNA averaged 1.0 +/- (SD) 0.33 relative densitometry units in controls (n = 4) and increased to 2.11 +/- 0.13 relative densitometry units in GH(3) rats (n = 5, p < 0.001). On the other hand, mRNA for the type I IGF receptor decreased in hypersomatotropic rats. Messenger RNAs for IGFBP-1 and IGFBP-4, which have been localized to renal tubules, both decreased significantly following growth induction, while IGFBP-3, the mRNA of which has an interstitial localization, was increased at week 10. CONCLUSION: These data suggest that there is a dynamic relationship between tubular and interstitial compartments with regard to the IGF system in the kidney which may be important in the regulation of the cell mass. PMID- 11474233 TI - Up-regulated TGF-beta mRNA expression in splenic T cells of high IgA-prone mice: a murine model of IgA nephropathy with glomerulosclerosis. AB - BACKGROUND/AIMS: Recently, we established a high serum IgA-prone inbred (HIGA) mouse strain as a murine model of spontaneous IgA nephropathy by selective mating of high serum IgA ddY mice, and found that they showed enhanced production of glomerular extracellular matrix components with increased expression of TGF-beta mRNA and protein in the kidneys. In this study, we examined the roles of lymphocytes in the development of high serum IgA in this strain. METHODS: We performed flow cytometric analyses of T and B cells in splenic mononuclear cells (SMNCs) from these mice using BALB/c mice as normal controls. We also compared serum TGF-beta1 concentrations and TGF-beta mRNA expression levels in the B-cell depleted (T-cell-rich) fraction of SMNCs in these mice. RESULTS: HIGA mice showed significantly fewer CD3-positive cells compared with BALB/c mice when young, but not when aged. The CD4/CD8 ratio of HIGA mice was lower than that of BALB/c mice, but this difference was not significant. Although the number of B220-positive cells did not vary significantly, the ratio of surface IgA-positive B cells was significantly increased in both young and adult HIGA mice. The B-cell-depleted SMNCs from HIGA mice exhibited higher levels of expression of TGF-beta and TGF beta1 mRNA than controls from a young age, which were maintained throughout life, but there were no differences in PDGF, MCP-1 or bFGF expression between these two strains. In contrast to local mRNA expression, serum TGF-beta1 concentration was decreased in HIGA mice compared with BALB/c controls. CONCLUSION: These findings suggest that the mating procedure performed to establish HIGA mice selected for a unique phenotype of local up-regulation of TGF-beta production in the kidneys, as well as T cells that may contribute to both the early and consistently high serum IgA expression and enhanced production of renal extracellular matrix components in HIGA mice. Additionally, TGF-beta1 may act locally, not systemically, in a paracrine or autocrine manner. PMID- 11474234 TI - Determination of bisphenol a in effluents of hemodialyzers. AB - This study was designed to determine whether bisphenol A (BPA) is eluted from hemodialyzers in which polycarbonate and polysulfone based on BPA are used as materials. Four types of polysulfone hemodialyzer (PS hemodialyzer: PS-A, PS-B, PS-C and PS-D) and an ethylene-vinyl alcohol copolymer dialyzer (EVAL hemodialyzer) were used in this study. In the PS-C, PS-D and EVAL hemodialyzers, polycarbonate was used in the case headers at both ends of the hemodialyzer. In in vitro experiments, the hemodialyzers were filled with reverse osmotic water, and BPA concentrations were measured. Saline solution (200 ml) was then circulated through a blood circuit tube connected to the hemodialyzer, and BPA concentrations in the saline solution were measured. In in vivo experiments, BPA concentrations in whole blood samples from hemodialysis patients treated with PS C (n = 3) and PS-D (n = 3) hemodialyzers were measured. In in vitro experiments, BPA was detected in the effluents of the PS-C, PS-D and EVAL hemodialyzers. In in vivo experiments, BPA was detected in whole blood samples from hemodialysis patients treated with the PS-D hemodialyzer (mean value, 0.77 ppb). This is the first report of BPA elution from hemodialyzers in which polycarbonate and polysulfone are used, and also the first report of detection of BPA in whole blood samples from patients on hemodialysis. PMID- 11474235 TI - Are uraemic children immunologically compromised? AB - BACKGROUND: Various immunological abnormalities leading to impaired immune status have been described in uraemic adults; however, few data are available for uraemic children. METHODS: In this study, peripheral blood total lymphocyte count and lymphocyte subsets (CD3+, CD4+, CD8+, CD16+, CD20+) were evaluated, skin tests with PPD and Candida antigens were performed, and serum immunoglobulin (IgG, IgA, IgM) and complement (C3, C4) levels were measured in 30 children with end-stage renal failure (10 before dialysis, 10 on continuous ambulatory peritoneal dialysis, and 10 on haemodialysis) and the results compared with those of 15 healthy controls. RESULTS: The data showed significant lymphopenia in predialysis and haemodialysis groups. No significant change was observed in the CD4+/CD8+ ratio or in the percentages of lymphocyte subsets in either group studied, while the absolute values of some lymphocyte subsets were significantly lower in all groups as compared with controls. In skin test evaluation, only the patients in the predialysis group showed a significantly decreased response to Candida antigen. The serum immunoglobulin levels were significantly decreased in the continuous ambulatory peritoneal dialysis group as compared with the control group. CONCLUSION: Our results, together with those of other paediatric studies, reported in the literature, suggest that uraemic children are not immunocompromised, though the effects of uraemia may cause some variation in their immune status. PMID- 11474236 TI - Retinopathy, hematuria, and diabetic nephropathy. AB - We have retrospectively analyzed the incidence of diabetic nephropathy in 21 diabetic patients who underwent renal biopsy between 1985 and 1995 for microscopic hematuria and/or proteinuria >2.5 g/day without retinopathy. Diabetic nephropathy was observed in 13 of 21 patients (62%). 50% of our patients with diabetic nephropathy had hematuria, the incidence being higher in type I as compared with type II diabetic patients (30 vs. 20%). Diabetic nephropathy without retinopathy but with hematuria was noted in 5 of 13 patients, and diabetic nephropathy without retinopathy and hematuria was also noted in 5 of 13 patients. We suggest from our retrospective analysis that renal-retinal diabetic syndrome really exists. PMID- 11474237 TI - Relationship between insulin resistance and uremic toxins in the gastrointestinal tract. AB - The relationship between insulin resistance and local uremic toxins was examined using an oral adsorbent. Fourteen rats demonstrating a diabetic state underwent two-thirds, nephrectomy and were divided into two groups. The control group was fed standard rat chow, and the test group was fed standard rat chow containing 5% AST-120. The target level of blood glucose was achieved by controlling the dosage of exogenous insulin. All rats were sacrificed at week 6. Body weight, blood glucose level, and renal function at week 6 were not significantly different between both groups. However, the mean blood glucose level and the mean dose of exogenous insulin in the AST-120-fed group were significantly reduced as compared with the control group. The results of the present study indicate that administration of an oral adsorbent in diabetic nephropathy decreases the doses of exogenous insulin and improves insulin resistance, and that uremic toxins which exist in the gastrointestinal tract play important roles. PMID- 11474238 TI - SDS-PAGE of urinary proteins for detection of nonglomerular hematurias. PMID- 11474239 TI - Decreased apolipoprotein levels are associated with decreased complement levels in acute glomerulonephritis. PMID- 11474240 TI - Secondary hyperparathyroidism, proinflammatory cytokines and response to epoietin in anemic maintenance dialysis patients. PMID- 11474241 TI - Clinicopathological study of early adenocarcinoma of the gastric cardia: comparison with early adenocarcinoma of the distal stomach and esophagus. AB - To clarify the clinicopathological characteristics of adenocarcinoma of the gastric cardia (AGC), including its association with Barrett's esophagus and intestinal metaplasia, 49 surgically resected early AGCs (EAGCs) were examined clinicopathologically, histopathologically, histochemically, and immunohistochemically. The clinicopathological characteristics of the patients with EAGC were compared with those of 293 patients with early adenocarcinoma of the distal stomach (EADS) and 7 patients with early adenocarcinoma of the esophagus (EAE). Histochemical staining with paradoxical concanavalin A (ConA) and immunohistochemical staining with monoclonal antibodies 45M1, Ccp58, and 56C6 were performed to investigate the differentiation phenotype of the tumor. ConA and 45M1 were used for markers of the gastric phenotype, and Ccp58 and 56C6 were used for markers of the intestinal phenotype. EAGC was associated with a higher mean age (p < 0.0001), a higher male-to-female ratio (p < 0.05), a higher incidence of elevated-type tumors (p < 0.0001), a higher incidence of differentiated-type tumors (p < 0.0001), and greater depth of invasion (p < 0.05) compared with EADS. EAE was associated with a higher incidence of elevated-type tumors (p < 0.001), a higher incidence of differentiated-type tumors (p < 0.05), and larger tumor size (p < 0.05) compared with EADS. The prevalence of Barrett's esophagus in patients with EAGC was significantly lower than in patients with EAE (10.2%, 5/49 patients vs. 100%, 7/7; p < 0.0001). The prevalence of intestinal (Barrett's) metaplasia in surrounding non-neoplastic mucosa in patients with EAGC was significantly lower than in patients with EADS or EAE (36.7%, 18/49 patients vs. 72.0%, 211/293 and 85.7%, 6/7; p < 0.0001 and p < 0.05, respectively). EAGC was associated with a higher incidence of tumors that reacted positively for gastric phenotype markers alone than EADS (32.7%, 16/49 cases vs. 17.1%, 50/293; p < 0.05) and a lower incidence of tumors that reacted positively for both gastric and intestinal markers than EADS or EAE (40.8%, 20/49 cases vs. 59.7%, 175/293 and 85.7%, 6/7; p < 0.05, respectively). Our findings indicate that AGC forms a specific category different from both adenocarcinoma of the distal stomach and esophagus in terms of association with Barrett's esophagus or intestinal metaplasia, and the differentiation phenotype of the tumor. PMID- 11474242 TI - Renal vein and vena cava involvement does not affect prognosis in patients with renal cell carcinoma. AB - OBJECTIVES: The prognostic value of tumor extension into the renal vein or vena cava is still a controversial issue. The aim of this study is to report our experience with radical surgery in patients with renal cell carcinoma (RCC) extending into the renal vein or subdiaphragmatic vena cava. METHODS: We evaluated 142 patients with RCC involving the renal vein or inferior subdiaphragmatic vena cava. RCC had extended into the renal vein in 118 patients and into the inferior vena cava in the remaining 24. Radical nephrectomy was performed in all cases with renal vein invasion. Radical nephrectomy with cavotomy and tumor thrombus removal was carried out in all cases with inferior subdiaphragmatic vena caval invasion. Cause-specific survival was calculated by means of the Kaplan-Meier method. The log rank test was used for survival comparisons and univariate analysis. RESULTS: The 5- and 10-year cause-specific survival rates were 51.5 and 39%, respectively, in the group of patients with tumor extension into the renal vein and 33.4% in those with inferior vena caval involvement. In 52 patients (44%), RCC extended only into the renal vein. In the remaining 66 patients, renal vein invasion was associated with other adverse prognostic factors. Life expectancy was lower for patients with other concurrent adverse prognostic factors than for those affected by renal vein involvement alone (p < 0.0001). In the latter group, survival expectancy was similar to those with stage T2N0M0 tumor. In 7 cases (29%), inferior vena caval invasion was not associated with other adverse prognostic factors. In the remaining 15 patients (71%), vena caval involvement was associated with other adverse prognostic factors. Concurrence of other adverse prognostic factors with vena caval invasion significantly decreased the disease-specific survival expectancy in comparison with the patients in whom vena caval involvement was the main prognostic factor (p = 0.008). In these patients, disease-specific survival was similar to those with stage T2N0M0 tumor. CONCLUSION: Renal vein or inferior subdiaphragmatic vena caval involvement does not significantly affect prognosis in patients with RCC. PMID- 11474243 TI - Complete response of highly advanced gastric cancer with peritoneal dissemination after new combined chemotherapy of S-1 and low-dose cisplatin: report of a case. AB - TS-1(S-1) has been developed as a new oral anticancer drug based on the biological modulation of 5-fluorouracil. We treated a patient with highly advanced gastric carcinoma with a new combination chemotherapy of S-1 and low dose cisplatin. Remarkable tumor reduction was observed after two cycles of this therapy in the primary tumor and metastatic lymph nodes, and the ascites disappeared. This was concluded to be a partial response. The only adverse effect was skin pigmentation of the fingers (grade 1), leading to early timing of operation after chemotherapy. The gastric tumor showed evident invasion to the serosa. Lymph nodes around the stomach were swollen. Peritoneal dissemination was also recognized in the omentum and mesocolon. Total gastrectomy with regional lymph node dissection was performed. Disseminated tumors were all resected. Histological examination showed that no tumor cells were detected in the gastric primary lesion, metastatic lymph nodes or disseminated peritoneal tumors, suggesting pathological complete remission. It was suggested that this regimen could be a potent combined therapy for the treatment of patients with highly advanced gastric carcinoma, and it could be useful as neoadjuvant chemotherapy. Further studies are necessary to evaluate the efficacy of this therapy. PMID- 11474244 TI - Successful treatment of bronchiolitis obliterans organizing pneumonia with low dose methotrexate in a patient with Hodgkin's disease. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is a rare disease, which is histopathologically defined by the presence of granulation tissue in the bronchioles, alveolar ducts and alveoli leading to plugging of the bronchiolar and alveolar lumen. BOOP is considered as a nonspecific response to many types of lung injury, including drugs, radiation, an underlying hematologic malignant neoplasm, autoimmune diseases, bacterial or virus infection, or an underlying lung disease, or occurs idiopathically. BOOP is mainly treated with corticosteroids, which induce a rapid clinical improvement. A frequent problem is relapse of disease when corticosteroid dosage is tapered off. We present the case of a 20-year-old patient with Hodgkin's disease developing BOOP after chemotherapy (COPP/ABVD) and irradiation. Initially, she responded well to corticosteroids, but relapsed when medication was discontinued. Complete remission of BOOP was achieved by long-term treatment with low-dose methotrexate (5-20 mg/week, i.v.). PMID- 11474245 TI - 5-fluorouracil administered as a 48-hour chronomodulated infusion in combination with leucovorin and cisplatin: a randomized phase II study in metastatic colorectal cancer. AB - OBJECTIVE: The primary objective of this trial was to determine the objective response of two regimens with CDDP administered every 2 weeks immediately before or after an 'optimal' 48-hour chronomodulated infusion of 5-fluorouracil (5-FU) modulated with leucovorin (LV) in metastatic colorectal cancer patients. Secondary endpoints were toxicity, 5-FU and its metabolites, plasma pharmacokinetics and progression-free and overall survival. METHODS: Metastatic colorectal cancer patients with measurable disease who were chemotherapy-naive or pretreated only with a 5-FU-bolus-based chemotherapy were eligible for this study. The study was designed as a randomized phase II clinical trial. RESULTS: Eighty-three patients were entered into the study. Forty-two were randomized to CDDP given before 5-FU and 41 to CDDP given after 5-FU. Patient characteristics were similar among the two groups. Toxicities were also similar among the two arms and the most frequent WHO grade III-IV toxicities were stomatitis (14%) and neutropenia (39-50%). Plasma pharmacokinetic profiles of 5-FU and 5-FUH2 were not significantly affected by the sequence of CDDP and 5-FU administration. Antitumor activity was similar in the two arms and was very promising both in pretreated patients (response rate 29%; 95% confidence interval 15-46%) and in chemotherapy naive patients (response rate 56%, complete response 9%, 95% confidence interval 40-71%). Median survival of the patients with and without pretreatment was 12 and 16 months, respectively. CONCLUSIONS: These results do not suggest a sequence dependence of the synergism between CDDP and 5-FU. However, they challenge the need of oxaliplatin to improve 5-FU/LV activity in advanced colorectal cancer. In fact, our results with an 'optimal' 5-FU dose and scheduling are very similar to those obtained with oxaliplatin plus 5-FU/LV. However, only a randomized phase III study will be able to give an answer to the hypotheses raised by this study. PMID- 11474246 TI - Oxaliplatin in combination with infusional 5-fluorouracil and leucovorin every 2 weeks as first-line treatment in patients with advanced colorectal cancer: a phase II study. AB - PURPOSE: To evaluate the efficacy and safety of oxaliplatin (L-OHP) in combination with leucovorin (LV)-modulated bolus plus infusional 5-fluorouracil (5-FU; de Gramont schedule) every 2 weeks in chemotherapy-naive patients with advanced colorectal cancer (CRC). PATIENTS AND METHODS: Thirty-two patients (median age: 69 years) with histologically confirmed and two-dimensionally measurable metastatic CRC were enrolled. The patients' performance status (WHO) was 0 in 14 (44%), 1 in 15 (47%), and 2 in 3 (9%) patients. Twenty (62.5%) patients had at least two metastatic sites. LV was administered at a dose of 200 mg/m2/day as a 2-hour intravenous infusion, followed by 5-FU as an intravenous bolus at the dose of 400 mg/m2 and then, as a 22-hour continuous infusion at the dose of 600 mg/m2/day for 2 consecutive days. L-OHP was administered on day 1 at the dose of 85 mg/m2 as a 2-hour infusion in parallel with LV but using different infusion lines. Treatment was administered every 2 weeks. RESULTS: In an intention-to-treat analysis, 2 (6.2%) complete and 9 (28%) partial responses (28%; odds ratio 34.2%; 95% confidence interval 17.92-50.83%) were achieved while 8 (25%) patients had stable disease and 13 (41%) progressive disease. The median duration of response was 5 months, but the median time to progression has not yet been reached. After a median follow-up period of 11 months, the median survival has not yet been attained, but the projected probability for 1-year survival was 72%. Grade 3/4 neutropenia occurred in 16 (50%) patients while 1 (3%) of them developed febrile neutropenia. There was no treatment-related death. Peripheral neuropathy grade 2 and > or =3 occurred in 5 (16%) and 7 (21%) patients, respectively. CONCLUSIONS: The bimonthly administration of L-OHP in association with LV-modulated bolus plus infusional 5-FU ('de Gramont' regimen) is a well tolerated and effective front-line treatment for metastatic CRC. PMID- 11474247 TI - Pilot study of albendazole in patients with advanced malignancy. Effect on serum tumor markers/high incidence of neutropenia. AB - Our preclinical studies have shown that the widely used antiparasitic drug albendazole has potent antiproliferative activity against colorectal cancer (CRC) and hepatocellular carcinoma (HCC). This trial was designed to evaluate albendazole in a small number of patients (n = 7) with either HCC or CRC and hepatic metastases refractory to other forms of therapy. Albendazole was given at 10 mg/kg/day orally in two divided doses for a period of 28 days. To follow the effect of treatment, tumor markers, carcinoembryonic antigen (CEA) or alpha-feto protein (AFP), were measured routinely in these patients. A range of hematological and biochemical indices were also serially measured to monitor bone marrow, kidney or liver toxicity. Albendazole therapy resulted in a decrease in CEA in 2 patients. In the remaining 5 with measurable tumor markers, serum CEA or AFP was stabilized in 3 patients, while in the other 2, after an initial stabilization (5-10 days), the markers began to increase. In the 7 patients completing the trial, albendazole was well tolerated and there were no significant changes in any hematological, kidney or liver function tests, but 3 patients were withdrawn for severe neutropenia which was probably contributory to the death of 1 patient. These data support our previous experimental results demonstrating that albendazole has antitumor effects. PMID- 11474248 TI - Clinical value of thymidine kinase in patients with cervical carcinoma. AB - OBJECTIVE: Our purpose was to determine the clinical value of thymidine kinase (TK), which is an important pyrimidine pathway enzyme involved in salvage DNA synthesis, in patients with cervical carcinoma. METHODS: We examined TK mRNA expression by reverse transcription polymerase chain reaction in 19 tissue specimens of invasive cervical carcinoma and 9 normal cervices and related it to thymidylate synthase (TS) and thymidine phosphorylase (TP) mRNA expressions. Serum TK level was determined by radioenzymatic assay in 79 patients with invasive cervical carcinoma, 7 patients with microinvasive carcinoma, 21 patients with carcinoma in situ and 32 normal women. RESULTS: TK mRNA expression was upregulated in invasive cervical carcinoma compared with the normal cervix (p < 0.05) and significantly correlated with TS mRNA expression (p < 0.0001) but not with TP mRNA expression. The serum TK level was significantly higher in patients with invasive carcinoma than in normal women and patients with carcinoma in situ (p < 0.01 and p < 0.05). In patients with invasive cervical carcinoma, the serum TK level significantly correlated with TK mRNA expression (p < 0.05), but not with any conventional clinicopathologic factors. High serum TK levels significantly correlated with a poorer survival (p < 0.05), and multivariate analysis showed serum TK level to be an independent prognostic factor (p < 0.05). CONCLUSION: TK may play an important role in influencing the malignant behavior of cervical carcinoma, and measurement of the serum TK level may be useful in predicting survival in patients with cervical carcinoma. PMID- 11474249 TI - Association between HLA-A/-B antigens and -DRB1 alleles and nasopharyngeal carcinoma in Tunisia. AB - Using serologic and molecular methods, 45 nasopharyngeal carcinoma (NPC) patients were typed for HLA class I and class II and were compared to 100 unrelated normal Tunisians. Our results showed that the antigen frequency of HLA-B13 and allelic frequencies of DRB1*03, DRB1*15 were significantly higher in the NPC patients than in the control group (15.5 vs. 4; 26.4 vs. 11.5, and 14.4 vs. 6.5%, respectively) probably indicating a positive association with NPC. Moreover, we observed that HLA-A23 was absent in our NPC sample and was present in 18% of normal controls, and HLA-DRB1*11 was less frequent among the patients compared to the controls (5.5 vs. 14%) suggesting a protective effect of this association with NPC. PMID- 11474250 TI - Functionally active duplications of the CYP2D6 gene are more prevalent among larynx and lung cancer patients. AB - The cytochrome P450 CYP2D6 is a polymorphic drug-metabolizing enzyme that is involved in the metabolism of several drugs and xenobiotics. Several independent studies indicate that the CYP2D6 metabolic status is a secondary factor in the risk of developing lung cancer, with individuals with high activity being at increased risk. The occurrence of functionally active duplications of the CYP2D6 gene is a phenomenon that affects 3-8% of Caucasians and up to 30% in some ethnic groups. These duplications cause ultrarapid metabolism of CYP2D6 substrates. In order to establish whether the highest CYP2D6 enzyme activity is associated with an increased risk of cancer, we analyzed the frequency of CYP2D6 gene duplications and enzyme-inactivating mutations in 199 Caucasian patients with lung or larynx cancer and in 335 healthy controls. A significantly increased frequency of carriers of the CYP2D6 gene duplication were found among lung and larynx cancer patients (13%), as compared with healthy controls (6.9%; p < 0.02). The frequency of the mutated active CYP2D6*9 allele was increased in lung cancer patients (p < 0.01) but not in larynx cancer patients. Global findings indicate that over 20% patients with lung or larynx cancer show CYP2D6 genotypes leading to ultrarapid metabolism or to the expression of an enzyme with altered kinetics (p < 0.01 vs. healthy controls). This may influence the metabolism of CYP2D6 substrates, including antineoplastic drugs and opioid derivatives used for pain relief in cancer patients. These patients would require higher doses than those considered as standard. We conclude that dosages for CYP2D6 substrates should be adapted to lung and larynx cancer patients. PMID- 11474251 TI - Clinicopathologic correlation of serum tissue polypeptide specific antigen in hepatocellular carcinoma. AB - OBJECTIVE: Recently, tissue polypeptide specific antigen (TPS) has been introduced as a cell proliferation marker. Little is known about its clinical significance in hepatocellular carcinoma (HCC). This study aimed to clarify serum TPS levels and tumor invasiveness of HCC. METHODS: Serum TPS levels were determined with a monoclonal TPS IRMA assay in 69 patients with HCC. A correlation between serum TPS levels and clinical, biochemical, and pathological features was sought and compared with that of alpha-fetoprotein (AFP). In 57 healthy subjects, 56 patients with biopsy-proven chronic hepatitis and in 49 patients with liver cirrhosis, serum TPS levels were assayed and compared. RESULTS: Serum TPS levels were significantly correlated with glutamic oxalacetic transaminase (p < 0.0001), glutamic pyruvic transaminase (p < 0.001), and lactate dehydrogenase (p = 0.027). There tended to be a positive relationship between serum TPS levels and tumor size, histological differentiation, capsular invasion, portal invasion, and clinical staging, although it did not reach statistical significance. A significant correlation, however, was observed between AFP and tumor size (p = 0.01), number (p = 0.042), histological grading (p = 0.028), portal invasion (p = 0.009), and clinical staging (p = 0.03). Patients with HCC had significantly higher TPS than healthy subjects (p < 0.001). However, there was substantial overlap between patients with HCC, chronic hepatitis, and liver cirrhosis. CONCLUSIONS: Our data suggest that serum TPS is not significantly related to tumor invasiveness in patients with HCC. Serum TPS levels are affected by the proliferative activity of the underlying chronic liver disease, which is frequently associated with HCC in Chinese patients. As a cell proliferation marker, serum TPS should be interpreted cautiously in the presence of chronic liver disease. PMID- 11474252 TI - Clinicopathological significance of cathepsin D expression in gastric adenocarcinoma. AB - OBJECTIVE: An estrogen-regulated lysosomal protease, cathepsin D, has been detected in a variety of tissues. This proteinase has been described as closely associated with tumor progression and metastasis in malignant tumors. The purpose of this study was to determine the clinicopathological and prognostic significance of cathepsin D expression in gastric adenocarcinoma. METHODS: In a consecutive series of 478 patients with gastric carcinoma (median follow-up period: 93 months, range: 1-285 months), cathepsin D expression in tumors was quantitatively analyzed with immunohistochemistry using a monoclonal antibody against cathepsin D (clone: 1C11). The percentage of cathepsin-D-positive cancer cells (the CD index) was calculated. In addition, the amount of cathepsin-D positive stromal cells was evaluated; three grades (high, intermediate, and low) were used for the classification. RESULTS: The mean CD index of 478 tumors was 12.8% (range: 0-100%, median: 8%). The mean CD index of diffuse-type gastric carcinomas (14.9%) was significantly higher than that of intestinal-type carcinomas (10.1%, p < 0.0001). Cathepsin D expression of cancer cells was significantly associated with the depth of tumor invasion in both types. The percentage of tumors with high cathepsin D expression in stromal cells was significantly higher in well-differentiated tumors (25.5%) than in moderately differentiated (12.8%) or in poorly differentiated tumors (19.1%). Cathepsin D expression of stromal cells was significantly associated with the depth of tumor invasion in the intestinal type, in contrast to the diffuse type. Highly expressed cathepsin D in cancer cells was associated with a poor prognosis in both types of carcinoma, but in stromal cells highly expressed cathepsin D was associated to a poor prognosis in the intestinal type only. CONCLUSION: These results indicate that cathepsin D expression in cancer cells may play an important role in tumor progression in diffuse-type gastric carcinoma, whereas in the intestinal type of carcinoma, cathepsin D expression in stromal cells may play an important role in tumor progression. PMID- 11474253 TI - Inoculation of human interleukin-17 gene-transfected Meth-A fibrosarcoma cells induces T cell-dependent tumor-specific immunity in mice. AB - OBJECTIVE: The biological activities of interleukin-17 (IL-17), a newly cloned cytokine, have not been fully elucidated. The present study was designed to assess the in vitro and in vivo effect of transfecting the IL-17 gene into tumor cells. METHODS: A complementary DNA (cDNA) encoding human IL-17 (hIL-17) was obtained by polymerase chain reaction amplification from the human CD4+ T cell cDNA library and inserted into the plasmid pRc/cytomegalovirus to construct an expression vector for the hIL-17 gene. Murine Meth-A fibrosarcoma cells were transfected with the hIL-17 gene using the lipofectin method. The hIL-17 gene expressing clone (Meth-A/IL-17) was selected and analyzed for cytokine expression by Northern blot. RESULTS: There was no significant difference in the in vitro proliferation rate among parent Meth-A, cells transfected with vector alone and Meth-A/IL-17 cells. When the tumor cells were transplanted subcutaneously into BALB/c nude (nu+/nu+) mice, there was no difference in in vivo growth rates among the three cell lines. Challenge with tumor cells in conventional BALB/c mice, however, resulted in the rejection of Meth-A/IL-17 cells, but the other two lines did grow. After immunization with Meth-A/IL-17 cells, the mice were rechallenged by parent Meth-A or syngeneic MOPC-104E plasmacytoma cells; the immunized mice rejected the Meth-A cells, but not the MOPC-104E cells. Injecting the anti-thy 1,2 (CD90), anti-CD4 or anti-CD8 monoclonal antibody into conventional BALB/c mice resulted in the resumption of in vivo growth of Meth-A/IL-17 cells, but injecting the anti-asialo GM1 antibody did not. Furthermore, flow cytometric analysis demonstrated a significant increase in the expression of major histocompatibility complex (MHC) class I and class II antigens and lymphocyte function-associated antigen-1 on Meth-A/IL-17 cells. CONCLUSION: Meth-A cells transfected with the hIL-17 gene can induce tumor-specific antitumor immunity by augmenting the expression of MHC class I and II antigens, and both CD4+ and CD8+ T cells may play important roles in inducing antitumor immunity, suggesting the possibility of developing a tumor vaccine incorporating IL-17-transfected tumor cells. PMID- 11474254 TI - CMF (cyclophosphamide, methotrexate, 5-fluorouracil) versus cnf (cyclophosphamide, mitoxantrone, 5-fluorouracil) as adjuvant chemotherapy for stage II lymph-node positive breast cancer: a phase III randomized multicenter study. AB - A multicenter phase III randomized study compared the efficacies of two adjuvant polychemotherapeutic regimens in 145 patients with stage II node-positive breast cancer. The standard chemotherapy combination, CMF (cyclophosphamide, methotrexate, 5-fluorouracil), was administered to 77 women. The experimental protocol, CNF (cyclophosphamide, mitoxantrone, 5-FU), in which mitoxantrone (Novantrone) replaced methotrexate, was given to 68 patients. Follow-up of the 145 patients by six participating hospitals showed no statistically significant difference (p = 0.6) between the two treatment regimens during a median follow-up of 4.5 years in terms of overall survival. There was, however, a significant advantage (p = 0.04) in the disease-free survival for those receiving mitoxantrone (mean survival 4.4 years for CNF versus 2.7 years for CMF). Toxic side effects associated with CNF (particularly alopecia and myelotoxicity) were relatively more frequent but acceptable and did not lead to dose reduction. In light of its association with improved disease-free survival in this study, larger studies should be undertaken on the role of mitoxantrone as adjuvant treatment in stage II breast cancer. PMID- 11474255 TI - Epirubicin--docetaxel combination in first-line chemotherapy for patients with metastatic breast cancer: final results of a dose-finding and efficacy study. AB - The aim of the study was to define a regular and tolerable dose of the epirubicin docetaxel combination in first-line chemotherapy of patients with metastatic breast cancer. Sixty-five women with measurable and/or evaluable disease were treated with epirubicin escalated from 60 to 110 mg/m(2) according to 5 dose levels, in combination with a fixed dose of 75 mg/m(2) docetaxel, every 21 days for 6 cycles, without preventive use of hematopoietic growth factors or antibiotics. Forty-three women received adjuvant chemotherapy, consisting of anthracyline- or anthracenedione-based regimens in 39 cases (60%). Twenty-seven women were treated in the phase I study (3 at epirubicin 60 mg/m(2), and 6 at each subsequent dose level). Dose-limiting toxicity consisted of grade III asthenia and febrile neutropenia (epirubicin 75 mg/m(2)), grade IV thrombopenia and grade III asthenia (epirubicin 90 mg/m(2)), grade IV stomatitis and grade III diarrhea (epirubicin 100 mg/m(2)), and grade III diarrhea (epirubicin 110 mg/m(2)). In the phase II study, an additional 38 women were treated at epirubicin 90 mg/m(2) and epirubicin 100 mg/m(2). During the 349 cycles delivered, grade IV neutropenia occurred in 90%; febrile neutropenia requiring hospitalization occurred in 62 (17.8%) and lasted more than 3 days in 12 (3.4%). Nonhematologic toxicity was acceptable. Three left ventricular ejection fraction depressions occurred and normalized during follow-up. The overall response rate in the 62 evaluable women was 69.4% (range: 58--81%), with a median duration of 7.8 months. After 26 months of follow-up, the median time to progression was 9.1 months and median overall survival was 22.7 months. On the basis of efficacy and toxicity, the recommended dose of the combination is epirubicin 100 mg/m(2) plus docetaxel 75 mg/m(2). PMID- 11474256 TI - Phase I clinical trial of weekly combined topotecan and irinotecan. AB - Combining antineoplastic analogues may increase efficacy by increasing the serum and intracellular concentration of the cytotoxic moiety shared by the analogues. Topotecan and irinotecan are two camptothecan analogues that are active in different human tumors (topotecan in ovary; irinotecan in colon) and in different experimental tumor systems. These data suggest that different mechanisms of drug resistance may be operative for the two agents, and if incomplete cross resistance exists between analogues, concomitant administration may be advantageous. The objectives of this phase I study were 1) to determine in a phase trial design whether topotecan and irinotecan administered concomitantly on a weekly schedule can be delivered at the same dose intensity as that of single agent topotecan or irinotecan delivery; and 2) to determine whether hematologic and/or nonhematologic toxicity is increased with topotecan and irinotecan administered together as a prelude to a possible phase II trial in responsive tumor categories. Irinotecan was administered for 30 to 45 minutes weekly x 4 at four dose levels: 50, 75, 100, and 125 mg/m(2)/wk. Topotecan was administered for 30 minutes (after irinotecan administration) at two dose levels within each of the irinotecan dose levels (1.0 and 1.5 mg/m(2)). Concomitant single-dose granulocyte-macrophage colony-stimulating factor (G-CSF) was used for leukocyte counts between 1,000 cells/mm(3) and 3,500 cells/mm(3) to maintain schedule. Maximum tolerated dosage (MTD) for the topotecan and irinotecan combination was defined as that which permitted 4 weeks of topotecan and irinotecan administration with G-CSF at or near the dose intensity reported for each single agent. Twenty-one patients received 32 4-week cycles. Dose-limiting toxicity was hematologic with grade IV leukopenia and neutropenia occurring at all dose levels. There was no apparent increase in the diarrhea syndrome associated with irinotecan. The MTD for irinotecan (at 125 mg/m(2)/wk) is the same MTD as with single-agent irinotecan use. The MTD for concomitant topotecan (1.5 mg/m(2)/wk) is 60% of the single-agent topotecan dose for the 5-day topotecan schedule (at 2.5 mg/m(2)/wk) but only 30% of the single-agent topotecan dose for the weekly schedule (5 mg/m(2)/wk). The topoisomerase I inhibitor dose is increased minimally when the analogues are administered concomitantly on a weekly schedule. Comparative trials of single-agent topotecan and irinotecan versus the combination of topotecan and irinotecan would be necessary to provide the proof of principle that combining analogues can increase therapeutic effectiveness. PMID- 11474257 TI - Parotid-sparing irradiation for cancer of the oral cavity: maintenance of oral nutrition and body weight by preserving parotid function. AB - The purpose of this study was to assess the nutritional benefit of parotid sparing irradiation, comparing the body weights of patients irradiated with parotid-sparing technique versus those irradiated with bilateral opposed photon beams, including both parotid glands in the radiation fields. One hundred fourteen patients with cancer of the oral cavity were irradiated with curative intent. Two-dimensional radiotherapy techniques sparing at least one parotid gland from the radiation beams were used to treat 31 patients (parotid-sparing techniques). Eighty-three patients were irradiated using bilateral opposed photon beams, which included both parotid glands (bilateral technique). Body weight during and after irradiation, treatment outcome, and survival were compared. Patients treated with parotid-sparing techniques maintained their nutritional intake and baseline body weight during and after irradiation. Patients treated with the bilateral technique that included both parotid glands had poor nutritional intake, leading to a more than 10% decline in their initial body weight; these patients did not regain their body weight during the 2-year follow up period. Primary tumor control rate was higher for patients treated with parotid-sparing techniques than for patients who had both parotid glands irradiated (70% versus 48%; p = 0.05). This difference is because a higher percentage of patients treated with parotid-sparing techniques had early-stage tumors (54%) compared with patients treated with the bilateral technique (24%). When analyzed according to the tumor stage, the primary tumor control rates for patients treated with parotid-sparing techniques and for patients treated with the bilateral technique showed no difference; control rates were 93% and 87% (p = 1.00) for early-stage tumors and 42% and 36% (p = 0.75) for advanced-stage tumors, respectively. Nodal control rates in the ipsilateral side of the neck and in the contralateral side of the neck for patients treated with parotid-sparing techniques and with the bilateral technique were not significantly different-74% versus 76% (p = 0.86) and 70% versus 82% (p = 0.21), respectively. Sparing at least one parotid gland during irradiation of patients with head and neck cancer will preserve parotid function and prevent xerostomia. Patients treated with parotid-sparing techniques were able to maintain their oral nutrition and body weight, compared with patients who had both parotid glands irradiated. A higher percentage of patients treated with parotid-sparing techniques had early-stage tumors, resulting in higher rates of primary tumor control and survival in this group of patients. PMID- 11474258 TI - Stevens-Johnson syndrome in a patient receiving anticonvulsant therapy during cranial irradiation. AB - A 28-year-old female patient with a recent history of breast carcinoma was referred to our clinic with generalized necrotic skin eruptions and severe mucosal erosions, which developed right after the completion of cranial radiotherapy for brain metastases. She had been receiving prophylactic diphenylhydantoin treatment 100 mg three times daily during radiation therapy. The extensive involvement of the oral mucosa with conjunctivitis and synechiae of the eyelids, facial swelling, and extension of the rash over the trunk and shoulders with bullous detachment of less than 10% of the total body surface strongly suggested Stevens-Johnson syndrome caused by phenytoin treatment in our patient. There has been conflicting evidence on the role of radiotherapy in the increased risk of severe drug reactions. Although various authors have emphasized the augmented rate of severe mucocutaneous reactions caused by anticonvulsants given during radiotherapy and suggested discontinuing the prophylactic use of such drugs in patients with no history of seizures, others have argued in favor of prophylactic anticonvulsants. Given the high risk of seizures, reaching 20% in patients with brain tumors, and the low incidence of drug reactions, the suggestion of refraining from prophylactic anticonvulsants in the setting of primary or metastatic brain tumors is controversial. PMID- 11474259 TI - Malignant proliferating trichilemmal tumor. AB - Proliferating trichilemmal tumors are uncommon neoplasms that are usually benign, but characterized by frequent local recurrence. In this report, we describe a patient who sought treatment at our clinic for a right occiput scalp nodule measuring 1 cm in diameter. The subcutaneous lesion was mobile and without overlying skin ulceration. Excisional biopsy of the mass was performed and the pathologic diagnosis returned as malignant, proliferating trichilemmal (pilar) tumor. Resection margins were free of tumor. On further examination the patient had no evidence of metastatic disease. A schedule of routine follow-up visits was arranged to monitor the site for recurrent disease. PMID- 11474260 TI - Combined regional and systemic chemotherapy by a mini-invasive approach for the treatment of colorectal liver metastases. AB - From February 1996 to December 1998, 95 patients affected with colorectal liver metastases underwent the positioning of an intraarterial hepatic catheter by a transcutaneous subclavian access, under local anesthesia. All patients were evaluated for catheter implantation complications. Moreover, 61 patients of 95 treated at our center were retrospectively evaluated for results of chemotherapy performed with two different schedules of hepatic artery infusion (HAI) combined with systemic chemotherapy (SC). Eleven patients (group A) were treated with combined SC (5-fluorouracil continuous infusion) and HAI (floxuridine). A subsequent 50 patients underwent HAI (floxuridine, 4 cycles) followed, if a response or stable disease were observed, by combined SC and HAI (group B). Three cases of aneurysm of subclavian artery occurred, which were treated by the positioning of a radiologic arterial stent and the reimplantation of the catheter by a femoral access. Thrombosis of the hepatic artery was registered in four cases. We observed 10.5% occurrence of dislocation of the catheter, which was always moved again in the hepatic artery. In group A, with 45% clinical objective response rate and 10% stable disease rate, median survival time and median time to extrahepatic progression were 9 and 6 months, respectively. In group B, we observed 44% clinical objective responses and 26% stable disease after HAI. Patients without disease progression and therefore submitted to sequential SC and HAI had a median survival time of 21 months and a median time to extrahepatic progression of 16 months. The development of the mini-invasive technique of implantation of an arterial port can avoid laparotomy for HAI. Percutaneous implantation of an arterial port has a low rate of technical complications. HAI followed by combined systemic and regional chemotherapy has good results in terms of survival and time to extrahepatic progression. PMID- 11474261 TI - Treatment of aggressive non-Hodgkin's lymphoma in elderly patients with thiotepa, Novantrone (mitoxantrone), vincristine, prednisone (TNOP). AB - The aging of the population and the increased incidence of non-Hodgkin's lymphoma will result in a large number of elderly patients with this disorder. Newer therapies will be required for this group of patients. This article reports a new therapy for elderly patients with diffuse aggressive non-Hodgkin's lymphoma. Patients were treated with TNOP (thiotepa 20 mg/m(2), mitoxantrone (Novantrone) 10 mg/m(2), vincristine (Oncovin) 1 mg/m(2) all on day 1 and prednisone 60 mg/m(2) on days 1 through 5 of a 21-day course. Twenty-six patients were enrolled on study. The patients' ages ranged from 66 years to 87 years, with a mean age of 75.5 years. Eleven patients had a partial response (42%) and 4 patients had a complete response (15%) for a total response of 57%. Eighty-one percent of patients survived 1 year and 54% survived for 2 years. The median survival was 26 months. Hematologic and nonhematologic toxicity was tolerable. We believe that TNOP is an excellent therapeutic option in this group of elderly patients, particularly in the palliative setting. PMID- 11474262 TI - Gastric metastases from breast cancer: a retrospective series of 12 patients. AB - Although well described in the literature, gastric metastases are often misdiagnosed in patients with breast cancer. The accuracy of diagnosis is critical because systemic therapy is beneficial, affording symptom palliation and an opportunity to avoid an unnecessary gastrectomy. PMID- 11474263 TI - A pilot study of pentoxifylline in the treatment of radiation-induced trismus. AB - Radiation-induced trismus has a significant impact on the quality of life of patients receiving radiotherapy for head and neck cancers. Pentoxifylline has immunomodulatory activities that downregulate certain cytokines that have been implicated as mediators of fibrogenic reactions after radiation. A pilot study was therefore conducted to evaluate the efficacy of pentoxifylline in the treatment of radiation-induced trismus. Twenty patients with severe trismus (dental gap < or = 25 mm) after receiving radiotherapy for nasopharyngeal carcinoma were enrolled onto the study. Four patients were excluded from analysis because of poor drug compliance. The remaining 16 evaluable patients completed an 8-week course of pentoxifylline at a dose of 400 mg two to three times daily. Changes in dental gap were recorded by measuring the distance between left upper and lower incisor before and after treatment. At the end of treatment, 10 patients had a measured increase in dental gap ranging from 2 to 25 mm. Six patients had an increment of 5 mm or more. The mean dental gap before treatment was 12.5 mm compared with 16.5 mm at the end of treatment (p = 0.023). The mean difference in dental gap was 4 mm, with a 95% confidence interval of 0.6 to 7.4 mm. The drug pentoxifylline appears to exert a modest therapeutic effect in patients with radiation-induced trismus, although our findings need to be confirmed by a randomized placebo-controlled study. While awaiting more evidence from clinical studies, a therapeutic trial of pentoxifylline is worth considering in patients experiencing radiation-induced trismus. PMID- 11474264 TI - Malignant fibrous histiocytoma of the trunk metastatic to the liver: treatment with transcatheter intrahepatic chemoembolization. AB - We report a patient with symptomatic liver metastases as the sole site of recurrence of a previously resected malignant fibrous histiocytoma of the trunk. We treated the patient with two sessions of transcatheter chemoembolization. Tumor regression was achieved, and the patient remained free of symptoms 23 months after the diagnosis of liver involvement, when he died of acute myocardial infarction. To our knowledge, this is the only case of transcatheter chemoembolization of liver metastases from soft-tissue sarcoma reported in the literature. Although it is a single case, it shows that this method may be a promising therapy for this grave disease. PMID- 11474265 TI - Combination of cisplatin-doxorubicin-cyclophosphamide in adenocarcinoma of unknown primary site: a phase II trial. AB - The purpose of this report is to evaluate toxicity, response, and survival of the cyclophosphamide-doxorubicin-cisplatin (CAP) chemotherapy regimen in patients with adenocarcinoma of unknown primary site (ACUP). Twenty-two patients with ACUP were eligible for this study between June 1992 and April 1999. There were 13 men (59%) and 9 women (41%) with a median age of 53.5 years (range: 29--78 years). Lung (seven), liver (six), vertebral bone site (six), and abdominal nodes (six) were the most common metastatic sites. Treatment consisted of doxorubicin 50 mg/m(2), cyclophosphamide 1,000 mg/m(2), and cisplatin 100 mg/m(2) (CAP), administered every 3 weeks; a total of six courses were planned. Twenty-two patients were assessable for toxicity and 20 patients were assessable for response. Grade III to IV neutropenia was observed in 14 patients (64%); febrile neutropenia occurred in 6 patients (27%) and in 10 cycles (12.5%). Grade III to IV anemia and thrombocytopenia were found in 12 (54.5%) and 9 patients (41%), respectively. Grade III to IV nausea and vomiting was observed in 9 patients (41%). Ten patients, 50% of the assessable population, obtained an objective response, including 3 complete (15%) and 7 partial (35%) responses. The median response duration was 3.9 months (range: 0.5--13.3 months). One patient (5%) had stable disease and 5 patients (25%) had progressive disease. The median overall survival and the median time to progression were 10.7 months (range: 0.4--56.9 months) and 8.8 months (range: 6.6--16.5 months), respectively. The CAP regimen in patients with ACUP had significant activity. This chemotherapy regimen induced a high level of grade III to IV toxicities and could not be considered as a treatment of reference. However, the emergence of long-term survivors among responder patients highlighted the need to search for an active treatment for patients with ACUP. PMID- 11474266 TI - Serum lactate dehydrogenase levels at presentation predict outcome of patients with limited-stage small-cell lung cancer. AB - Serum lactate dehydrogenase (LDH) is a biochemical parameter that is elevated in the majority of extensive-stage small-cell lung cancer (SCLC). In this study, distribution and prognostic importance of serum LDH in limited-disease SCLC were investigated. Serum concentrations of LDH were measured in 184 patients at initial examination. These results were compared with prospectively recorded clinicopathologic characteristics and patient outcome data. Significant positive association was found between LDH levels and weight loss, performance status, response to chemotherapy, and albumin but not between age, gender, and hemoglobin values. Patients with high concentrations of LDH had a significantly worse prognosis than did patients with normal levels. The probability of overall survival at 1 year was 60.2% in patients with normal serum LDH levels and 33.1% in patients with higher values (p = 0.0017). Also, the prognostic value of LDH on overall survival was shown in multivariate analysis (p = 0.05). At the time of diagnosis, serum levels of LDH appear to have a significant relation to outcome in patients with limited-stage SCLC. PMID- 11474267 TI - Amiodarone and radiation therapy sequelae. AB - With the aging of our population, amiodarone, a cardiac antiarrhythmic, is being used with increasing frequency. Anecdotal reports have appeared describing unexpected mucosal as well as cutaneous toxicity in patients treated concurrently with external beam radiotherapy and oral amiodarone. We retrospectively reviewed 12 years of experience at the University of Iowa Hospitals and Clinics and found 10 cases wherein amiodarone was taken concurrently with external beam radiation therapy. In our series, there were no missed treatment fractions because of unexpected acute sequelae. We conclude that amiodarone use does not preclude the delivery of therapeutic radiotherapy. PMID- 11474268 TI - Systemic chemotherapy in patients with renal failure. AB - The administration of systemic chemotherapy to patients with moderate-to-severe organ impairment remains a challenge. In renal failure, guidelines for dose adjustments exist for very few chemotherapy agents. Subsequently, oncologists typically withhold treatment with systemic chemotherapy for patients with severe renal dysfunction. We present the case of a patient with metastatic transitional cell carcinoma of the bladder and renal failure who underwent successful systemic chemotherapy with paclitaxel and carboplatin. The data on systemic chemotherapy in patients with severe renal dysfunction are reviewed. PMID- 11474269 TI - Palliative radiotherapy for synchronous bilateral lung cancers. AB - Scant information is available about the outcomes of patients with synchronous bilateral lung cancers (SBLC) treated with radiation. We report on 32 patients with SBLC who were treated for palliation by external beam irradiation during a 19-year period. The overall median survival was 7 months; 20 individuals (63%) survived at least 6 months. Among the evaluable patients, the complete/partial subjective and objective response rates were 100% and 60%, respectively. Survival was not significantly influenced by the total dose applied. Radiotherapy is effective for palliation of symptoms associated with SBLC. However, the dismal prognosis of SBLC managed by nonradical radiotherapy alone warrants investigation of treatment using curative doses of radiotherapy or chemotherapy with radiotherapy. PMID- 11474270 TI - Validity of local treatment including intraarterial infusion chemotherapy and radiotherapy for fungating adenocarcinoma of the breast: case report of more than 8-year survival. AB - A 39-year-old woman came to us complaining of severe anemia (hemoglobin: 2.3 g/dl) and a painful right breast that was entirely occupied by an ulcerative, foul-smelling tumor approximately 20 cm in maximum dimension. The tumor, which was determined to be mucinous adenocarcinoma at biopsy, had invaded the chest wall with multiple lung metastases (T4cN2M1 stage IV). After a blood transfusion, the patient received the following multimodal treatment: concurrent chemoradiotherapy (50 Gy), intraarterial infusion chemotherapy consisting of doxorubicin 50 mg, mitomycin-C 10 mg, and cisplatin 50 mg, and skin graft surgery. After intraarterial infusion chemotherapy, the fungating tumor disappeared. The patient experienced a relapse of right pleural effusion 2 years later and received multimodal treatment. Now, March 30, 2000, the patient is doing well without local recurrence, 8 years after her first admission. The combination of intraarterial infusion chemotherapy and radiotherapy plays a role in successful treatment of extensive local disease of the breast. PMID- 11474271 TI - Combined 5-fluorouracil infusion with fractionated epirubicin and cyclophosphamide in advanced breast cancer. AB - 5-Fluorouracil (5-FU) given by continuous infusion (c.i.) allows higher dose delivery, causes less myelosuppression, and may interfere with repair of DNA damage caused by epirubicin and cyclophosphamide. With this rationale, we conducted a phase II study to test the activity and toxicity of 5-FU c.i., epirubicin, and cyclophosphamide in patients with metastatic breast cancer (MBC). Twenty-eight patients with MBC were entered in the trial. 5-FU (200 mg/m(2)) was administered by c.i. from day 1 to day 20. Epirubicin (35 mg/m(2)) and cyclophosphamide (400 mg/m(2)) were administered from day 2 to day 4, every 4 weeks. All patients were evaluable for response and toxicity. A total of 125 courses of chemotherapy were administered, with a median of 4 per patient (range: 2--6). Toxicity, assessed using World Health Organization criteria, was as follows: nausea and vomiting grade III--IV occurred in 36%, alopecia (grade III) in 86%, neutropenia (grade III--IV) in 50%, and cardiac toxicity grade I--II in 11% of patients. Five patients (17.9%) had a complete response to therapy, and 16 (57.1%) had a partial response (response rate 75%, 95% CI 55--89%). Disease stability and progression occurred in 4 (14.3%) and 3 (10.7%) patients, respectively. Median time to progression was 13.1 months (range: 3.4--66.9+), and median survival time was 27.7 months (range: 5.4--67.1+). Outpatient treatment with combined 5-FU c.i., epirubicin, and cyclophosphamide shows high activity in advanced breast cancer and gives prolonged remission with acceptable toxicity. PMID- 11474272 TI - Paget disease of the breast: analysis of 41 patients. AB - The treatment for the patients with Paget disease of the breast is controversial. This review of its natural history, treatment approach, and clinical outcome will help to formulate treatment. Forty-one patients with a diagnosis of Paget disease of the breast were retrospectively reviewed at Providence Hospital & Medical Centers from 1980 to 1999. Ninety-eight percent of patients had underlying carcinoma (ductal carcinoma in situ and/or invasive ductal cancer). Patients with a palpable mass have a much higher incidence of invasive ductal cancer, positive lymph node, and a worse survival rate. The median length of follow-up was 42 months (range: 6--200 months). Twenty-seven percent of patients (11/41) had conservative operations, including 1 patient with a palpable mass; 10 patients with no palpable mass; and 3 patients with recurrence after conservative operation. Thirty-seven percent of patients received adjuvant therapy. Paget disease of the breast has very high incidence of underlying carcinoma (100% in a palpable mass, 96% in nonpalpable mass). Patients with a palpable mass have a worse survival than do patients with nonpalpable mass. Conservative operation should cautiously be selected even for patients with no palpable mass because of a higher recurrence rate. PMID- 11474273 TI - Pharmacologic study of paclitaxel administered with or without the cytoprotective agent amifostine, and given as a single agent or in combination with epirubicin and cisplatin in patients with advanced solid tumors. AB - The main purpose of this study was to investigate whether the coadministration of amifostine alters the pharmacokinetic behavior of paclitaxel. Eight patients were included in the study: six received paclitaxel in combination with epirubicin and cisplatin, and two received paclitaxel as a single agent. Doses of paclitaxel in these protocols were 135, 150, 175, and 200 mg/m(2) and two patients were treated at each dose level. Pharmacokinetic sampling for paclitaxel analysis was performed in each patient during two consecutive cycles, one with and one without amifostine (750 mg/m(2) as a 15-minute intravenous infusion 30 minutes before paclitaxel administration). At each dose level, the pharmacokinetic data of paclitaxel were compared per patient for a cycle without amifostine versus a cycle with amifostine. Amifostine did not seem to interact pharmacokinetically with paclitaxel, given either alone or in combination chemotherapy. This is in line with the clinical findings that amifostine has no negative effects on the antitumor activity of various antineoplastic agents. Also, amifostine may reduce toxic effects of combination chemotherapy regimens that include paclitaxel. PMID- 11474274 TI - A dose-finding study of the weekly administration of paclitaxel in patients with advanced solid tumors. AB - The purpose of this study was to determine the dose-limiting toxicities and the maximum-tolerated dose (MTD) of weekly administration of paclitaxel in patients with advanced solid tumors. Twenty-six patients with advanced solid tumors were treated with escalated doses of paclitaxel (starting dose 70 mg/m(2)/wk with increments of 10 mg/m(2)/wk) for 4 consecutive weeks every 6 weeks. No intrapatient escalation or growth factor support was allowed. The DLT was exceeded at the dose of 120 mg/m(2)/wk, and the dose-limiting events were grade IV neutropenia and treatment delay because of incomplete hematologic recovery. There was no cumulative myelosuppression. Grade IV neutropenia occurred in four (6%) cycles, and there was one episode of febrile neutropenia. Grade II/III fatigue occurred in 19 (73%) patients, resulting in discontinuation of treatment in 2 of them; grade II neurosensory toxicity and grade II alopecia occurred in 8 (31%) patients each. The MTD, which is also the recommended dose for further phase II studies, was 110 mg/m(2)/wk. Among the 21 patients with bidimensionally measurable disease, 2 (10%) partial responses were observed, both in patients with heavily pretreated advanced breast cancer. The weekly administration of paclitaxel for 4 consecutive weeks in cycles of 6 weeks is a feasible, safe, and active outpatient regimen that merits further evaluation in combination with other anticancer agents. PMID- 11474275 TI - Relapsed Wilms tumor: is there a role for radiation therapy? AB - From 1968 to 1996, 21 of 97 (22%) patients with Wilms tumor treated at the University of Iowa Hospitals and Clinics relapsed after initial therapy. The most common sites of initial recurrence were the lungs in 12 (57%), abdomen in 3 (14%), bone in 3 (14%), and liver in 2 (10%). Treatment for recurrence included chemotherapy in 16, radiotherapy in 15 and surgery in 8. Radiotherapy was given to bilateral lungs of 8 previously untreated chests; total dose was 1,200 cGy to 1,500 cGy, with 2 patients receiving 1,000-cGy boost. One patient had abdominal radiotherapy after surgery for relapse; six patients had palliative radiotherapy to distant sites. The 1-, 3-, and 5-year postrelapse survivals were 62 +/- 11%, 38 +/- 11%, and 33 +/- 10%, respectively. Using Cox proportional-hazard regression models, abdominal recurrence (p = 0.0002; hazard ratio, 39.5) and initial stage IV disease (p = 0.019; hazard ratio, 6.27) were found to have a negative impact on postrelapse survival. Seven patients with previously nonirradiated chests are alive at a median follow-up of 123 months after relapse (range, 55--272 months); all had whole lung irradiation with or without chemotherapy at relapse for lung metastases. Radiotherapy was also found to be an effective treatment in the palliation of painful metastases. PMID- 11474276 TI - Treatment of vinorelbine-associated tumor pain. AB - Vinorelbine is a synthetic vinca alkaloid used for the treatment of Hodgkin's disease and metastatic breast, non-small-cell lung, and ovarian cancers. Vinorelbine-associated tumor pain can be so severe that patients are unable to complete therapy. We present a series of three patients who were premedicated with ketorolac in an attempt to control infusion-related pain. Two of our patients responded well and were able to complete their courses of vinorelbine therapy. The third had persistently severe pain, and therapy was terminated because of pain and lack of response to vinorelbine treatment. These three cases support the hypothesis that surgery or radiation therapy before the use of vinorelbine may increase the likelihood of experiencing associated tumor pain. Premedication with ketorolac and morphine before vinorelbine infusion is proposed as a reasonable prophylactic treatment for associated tumor pain. PMID- 11474277 TI - Hypercalcemia mediated by parathyroid hormone-related protein as an early manifestation of pancreatic adenocarcinoma metastasis: a case report. AB - Humoral hypercalcemia of malignancy (HHM) is a paraneoplastic syndrome rarely associated with pancreatic adenocarcinoma. Parathyroid hormone-related peptide (PTHrP) is the central mediator of this condition. In our patient, hypercalcemia associated with elevated PTHrP was the initial manifestation of metastatic pancreatic adenocarcinoma. Successful palliation of HHM with bisphosphonates and loop diuretics has been previously reported and was effective in our patient. We report the first case of pancreatic adenocarcinoma metastasis after successful resection to present with hypercalcemia. PMID- 11474278 TI - Primary osteogenic sarcoma arising from the dura mater: case report. AB - A 42-year-old woman who sought treatment for left drop foot was found to have a right frontoparietal parasagittal mass. Gross total resection of the tumor was performed and pathologic analysis revealed high grade osteoblastic osteosarcoma. The patient received adjuvant chemotherapy and continues to do well with no evidence of metastases or local recurrence 3 years after initial presentation. PMID- 11474279 TI - Capecitabine for 5-fluorouracil-resistant brain metastases from breast cancer. AB - We report a case in which brain metastases originating from breast cancer responded to treatment with oral capecitabine. The metastases had progressed and Karnofsky performance status deteriorated despite whole brain irradiation, hormonal treatment, and systemic chemotherapy that included 5-fluorouracil (5 FU). In contrast, 2 months of treatment with oral capecitabine produced a partial response, documented by lesion size on magnetic resonance imaging and an improvement in performance status; both measures continued to improve during 11 months of capecitabine treatment. PMID- 11474280 TI - Is MUGA scan necessary in patients with low-risk breast cancer before doxorubicin based adjuvant therapy? Multiple gated acquisition. AB - Doxorubicin-based chemotherapy in the adjuvant treatment of breast cancer has become standard. Use of doxorubicin is limited by cardiac dysfunction; however, the incidence is dramatically reduced by limiting the dose to less than 550 mg/m(2). Although the cumulative dose in breast cancer is typically 240 mg/m(2), multiple gated acquisition (MUGA) scans are still recommended for determining cardiac functional status in these patients. To examine the need for this practice, we reviewed 296 patients who underwent surgery for breast cancer at Roswell Park Cancer Institute between July 1997 and December 1998. Fifty-nine of 95 (62%) patients receiving doxorubicin-based regimens, and 3 of 39 (7%) receiving nondoxorubicin regimens had pretreatment MUGA scans. The MUGA scans showed normal results in 58 patients and low-normal in 4 (6.5%), with no wall motion abnormalities encountered. There were no cases where doxorubicin was not used because of an abnormal MUGA scan. There were no cardiac complications in the 59 women who received doxorubicin-based chemotherapy. MUGA will screen out few, if any, women under consideration for doxorubicin-based adjuvant therapy; the decision to avoid doxorubicin can be made based on age and preexisting comorbidity. Guidelines recommending routine use of MUGA before the administration of doxorubicin for adjuvant therapy for breast cancer should be reconsidered. PMID- 11474281 TI - Gastrointestinal autonomic nerve tumor: immunohistochemical and molecular identity with gastrointestinal stromal tumor. AB - Gastrointestinal autonomic nerve tumor (GANT) is a gastrointestinal neoplasm that ultrastructurally recapitulates the enteric neural plexus. This study identifies and defines the features of 10 cases of this rare mesenchymal tumor and compares its clinicopathologic and molecular genetic features with the data on gastrointestinal stromal tumor (GIST). The majority of patients in this series presented at an older age (mean 64 years). Tumors arose from the stomach (6), small intestine (2), and retroperitoneum (2). Mean tumor size was 14 cm; however, four neoplasms were <6 cm. Histologically, tumors were spindled or epithelioid; one epithelioid tumor demonstrated a previously undescribed rhabdoid histologic phenotype. All tumors were positive for CD117 (KIT), while eight of 10 were positive for CD34. In contrast, only two were positive for S-100, and all were negative for actin and desmin. Five GANTs demonstrated GIST-specific gain-of function mutations in the juxtamembrane domain of the c-kit gene (50%). Three of 10 patients died of disease in 22-30 months, one patient died in the postoperative period, and one patient died of complications of CML. The clinicopathologic, histologic, immunohistologic, and molecular features of GANT are similar to GIST, indicating that GANT merely represents a phenotypic variant of GIST. PMID- 11474282 TI - Serous tumors involving extra-abdominal/extra-pelvic sites after the diagnosis of an ovarian serous neoplasm of low malignant potential. AB - The involvement of extra-abdominal/extra-pelvic sites by serous tumors after the diagnosis of an ovarian serous neoplasm of low malignant potential is extremely rare. In this study we present the clinicopathologic features of 12 such cases seen at our institution during a period of 19 years (1980-1999). The patients' age ranged from 19 to 50 years (mean 33 years). By FIGO staging the original ovarian tumors were distributed as follows: stage I, 4; stage II, 2; stage III, 5; unknown stage, 1. All patients were treated surgically. Ten patients also received adjuvant therapy (radiotherapy, 2; chemotherapy and radiotherapy, 4; chemotherapy, 3; intraperitoneal 32P, 1). The interval between the diagnosis of the ovarian neoplasm and the subsequent tumor involving an extra-abdominal/extra pelvic site ranged from 4 to 240 months (mean 124 months). Sites of extra abdominal/extra-pelvic involvement and the number of cases were as follows: left neck lymph nodes (LNs), 4; left and right neck LNs, 1; pleura, 2; lung, 1; mediastinum, 1; chest wall, 1; axillary and chest LNs, 1; and vertebral body, 1. Eight patients were treated with chemotherapy, 1 with radiotherapy, 2 with chemotherapy and radiotherapy, and 1 with surgery alone. Follow-up ranging from 5 months to 18 years was available in 11 patients. Six patients died of disease and 5 patients were alive with no evidence of disease. In this small series of cases, no definitive clinical or pathologic feature related to the occurrence of extra abdominal/extra-pelvic serous tumors was found. Based on the LN involvement and the endosalpingiosis seen in some cases, these tumors might develop from circulating neoplastic serous cells or from areas of endosalpingiosis involving extra-abdominal/extra-pelvic sites. PMID- 11474283 TI - Pathologic and clinical features of primary pulmonary extranodal marginal zone B cell lymphoma of MALT type. AB - We reviewed pathologic, phenotypic, and clinical features of extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) type primarily involving lung to address unresolved questions regarding behavior and pathologic features of unambiguously diagnosed pulmonary MALT lymphoma. Lung specimens from 50 patients were reviewed. Forty-one had low-grade MALT lymphoma. Nine had low grade MALT lymphoma and diffuse large B-cell lymphoma. The patients included 32 women and 18 men with a median age of 68 years (range 34-88 years). Half of the patients were asymptomatic at the time lymphoma was diagnosed. Radiographic abnormalities were more commonly unilateral (37 patients) than bilateral (12 patients). Localized masses or nodules occurred in 39 patients. Associated autoimmune disorders (29%) and monoclonal gammopathies (43%) were common. Low grade lymphomas formed intraparenchymal masses composed of centrocyte-like cells, plasmacytoid lymphocytes, and plasma cells that formed lymphoepithelial lesions and exhibited a lymphangitic growth pattern. Mediastinal lymph nodes were involved histologically in 44% of cases. Lymphoma-specific survival was 71.7% at 10 years, and overall survival was significantly worse than age-and gender matched control patients. None of the following features predicted those patients who had an adverse outcome: systemic symptoms, presence of autoimmune disorders or paraproteinemia, anatomic distribution and number of pulmonary lesions, lymph node involvement, or presence of anthracycline-treated large B-cell lymphoma. PMID- 11474284 TI - Low-grade (fibromatosis-like) spindle cell carcinoma of the breast. AB - Spindle cell carcinoma of the breast, a variant of metaplastic carcinoma, includes a wide spectrum of lesions with histomorphologic and nuclear features ranging from overtly malignant to mildly atypical. Spindle cell carcinomas with mildly atypical features may resemble fasciitis, fibromatosis, or myofibroblastic tumors and therefore are often misinterpreted as such. A recent study has suggested that spindle cell carcinomas with a dominant fibromatosis-like phenotype, unlike spindle cell carcinomas in general, have no propensity for distant metastasis and should be termed "tumors" rather than "carcinomas." To investigate the question of fibromatosis-like spindle cell breast carcinoma (FLSpCCs) metastatic potential, we studied cases of FLSpCC seen at the University of Texas M.D. Anderson Cancer Center between 1987 and 2000. Clinical, pathologic, and immunophenotypic features were reviewed, with emphasis on biologic behavior and predictors of clinical outcome. Our series included 24 women who ranged in age from 55 to 85 years (mean 66 years). Tumor size ranged from 1.0 to 5 cm (mean 2.8 cm). Most tumors were grossly well defined but had microscopic infiltrative borders. Tumors showed a dominant fibromatosis-like or myofibroblastic-like growth pattern with prominent collagenization. Inflammatory infiltrate was noted in the majority of tumors. Cytokeratin-positive cells were seen in all cases and usually appeared as cords or sheets of polygonal cells; isolated cytokeratin positive cells were rare. In most tumors immunoreactivity for smooth muscle actin (SMA) was confined to the cytokeratin-negative cells. In five cases intense co expression of cytokeratin and SMA was noted. None of the tumors showed immunoreactivity for smooth muscle heavy chain myosin, estrogen receptors, progesterone receptors, or HER-2/neu. Ki-67 expression was noted in fewer than 5% of tumor cells. Treatment consisted of local excision (seven cases) or modified radical mastectomy (13 cases). Treatment was unknown in four cases. In patients who underwent axillary nodal dissection, no lymph node metastases were found. Two of the six patients who underwent local excision developed local recurrence. Two patients who underwent modified radical mastectomy developed lung metastases within 2 years after the initial diagnosis. The metastatic tumors were histologically similar to the primary tumors. Our findings indicate that FLSpCCs have the potential for local recurrence and distant metastasis and should be treated accordingly. Because FLSpCCs may be underdiagnosed as benign, the use of immunohistochemical studies, especially for cytokeratins and SMA, is essential in the evaluation of any spindle cell proliferations of the breast. PMID- 11474285 TI - Core biopsy of the breast with atypical ductal hyperplasia: a probabilistic approach to reporting. AB - The diagnosis of atypical ductal hyperplasia (ADH) at needle core breast biopsy (NCB) is typically regarded as an indication for surgical excision. Although ADH is an intermediate risk nonobligate precursor lesion, the rationale for further therapy is the result of a reported high prevalence of a concomitant more advanced lesion (typically ductal carcinoma in situ) as the index lesion. To assess whether certain histopathologic features of ADH in NCB are predictive of open biopsy outcomes, the authors correlated the extent and pattern of ADH in 47 core biopsies (11-or 14-gauge) with the subsequent surgical specimen. Extent of ADH on NCB was ascertained by determining the number of large ducts and/or terminal duct-lobular units affected, with involvement of one large duct or one terminal duct-lobular unit representing a single focus, involvement of one duct and one terminal duct-lobular unit as two foci, and so on. Of the 47 cases, ADH was restricted to < or =2 foci in 24 cases (51.1%), confined to 3 foci in 8 cases (17.0%), and involved > or =4 foci in 15 cases (31.9%). The corresponding histopathologic findings at excision were benign lesions without atypia (n = 14), focal residual ADH (n = 13), atypical lobular hyperplasia (n = 3), ductal carcinoma in situ (n = 15), and invasive mammary carcinoma (n = 2). When the number of foci of involvement by ADH on NCB (based on an average of 11.6 cores per case) was correlated with the open biopsy results, all cases of ADH limited to < or =2 foci had no worse lesion on excision, whereas ADH present in > or =4 foci was found to be a strong predictor of a more advanced lesion on excision (p <0.0001, chi2). When histologic pattern was evaluated, all cases of pure micropapillary ADH on NCB showed pure micropapillary ductal carcinoma in situ on excision. PMID- 11474286 TI - Mammary-type myofibroblastoma of soft tissue: a tumor closely related to spindle cell lipoma. AB - Mammary myofibroblastoma is a benign breast tumor, with a reported predilection for older men. It is composed of fascicles of spindle cells having features of myofibroblasts, with intervening hyalinized collagenous stroma and a variably prominent component of adipose tissue. The spindle cells characteristically express both CD34 and desmin. Herein, we report the clinicopathologic features of nine tumors that were morphologically and immunohistochemically identical to myofibroblastoma of breast; however, they arose in subcutaneous soft tissue at extramammary sites. The study group comprised seven men and two women with an age range of 35-67 years (median 53 years). Lesions presented as either a slowly growing painless mass or were incidental findings at the time of surgery. The site distribution was as follows: inguinal/groin area (five cases) and one case each in posterior vaginal wall, buttock, anterior abdominal wall, and mid-back. Tumor size ranged from 2 to 13 cm (median 6 cm), and all lesions were well circumscribed. Eight tumors had a component of adipose tissue (ranging from 10% to 60%), within which some variation in adipocyte size was often seen. One case showed epithelioid cytomorphology and three cases showed rare atypical or multinucleated cells. Focal myxoid stromal change was seen in four cases. Tumor cells were positive for desmin (9 of 9 cases), CD34 (8 of 9 cases), and occasionally positive for smooth muscle actin (3 of 9 cases). Lesions were marginally excised with no recurrences to date, although follow-up is very limited. Lesions with morphologic and immunophenotypic features similar to myofibroblastoma of breast can arise at extramammary sites, with an apparent predilection for the inguinal area of older men. Both mammary and extramammary lesions show morphologic overlap with spindle cell lipoma and are likely closely related. PMID- 11474287 TI - Pleomorphic leiomyosarcoma: clinicopathologic and immunohistochemical study with special emphasis on its distinction from ordinary leiomyosarcoma and malignant fibrous histiocytoma. AB - Pleomorphic leiomyosarcoma (PLMS) was recently described as a morphologic variant of leiomyosarcoma; however, its diagnostic criteria, as shown by morphologic features and biologic behavior, remain controversial. We describe 28 cases of pleomorphic sarcoma with pleomorphic areas in more than two thirds of the tumor and an ordinary leiomyosarcomatous fascicular area covering less than one third as PLMS. PLMS comprised 8.6% of all the leiomyosarcomas (322 cases) registered in our institute. Patients ranged in age from 31 to 89 years (average, 57.9 years). Seventeen patients (60.7%) were male and 11 were female. Tumor location was as follows: the extremities in 17 cases, the retroperitoneum or abdominal cavity in 7 cases, the chest/abdominal wall in 3 cases, and the scalp in 1 case. Histologically, all cases showed at least small foci of fascicles consisting of smooth muscle tumor cells, in addition to pleomorphic areas mimicking storiform pleomorphic malignant fibrous histiocytoma. The border between pleomorphic and leiomyosarcomatous fascicular areas was sharp in 3 cases, gradual in 2 cases, and blending in 23 cases. Sixteen cases (57.1%) showed a typical storiform pattern, 6 cases revealed extensive stromal hyalinization, 6 cases showed a chronic inflammatory infiltrate, 2 cases had the foci of foamy xanthomatous cells, and 7 cases contained myxoid malignant fibrous histiocytoma-like areas covering less than 50% of the tumor. The tumors had a tendency to be of a morphologically higher grade (10 tumors were French Federation of Cancer Centers grade 2, 18 were grade 3). Five of 28 cases (18%) showed rhabdoid features. Immunohistochemically, all of the 28 tumors examined showed a positive reactivity for at least one smooth muscle marker (desmin, muscle-specific actin, and alpha-smooth muscle actin) in the leiomyosarcomatous fascicular areas. In the pleomorphic areas the expression of smooth muscle markers (desmin 10 of 28, muscle-specific actin 13 of 28, and alpha-smooth muscle actin 14 of 28) was significantly reduced, compared with that in leiomyosarcomatous fascicular area (desmin 18 of 28, muscle-specific actin 26 of 28, and alpha-smooth muscle actin 24 of 28). No significant difference was observed between the MIB-1 labeling index in the leiomyosarcomatous fascicular areas (26.10 on average) and that in the pleomorphic areas (26.17 on average). However, the MIB-1 labeling index in PLMS was significantly higher than that in ordinary leiomyosarcoma (n = 20, 12.86 on average) or storiform-pleomorphic malignant fibrous histiocytoma (n = 16, 16.63 on average). In 23 patients follow-up data were available with a duration of 1 239 months. Eleven patients developed metastases, and lung accounted for the most common site of metastasis (9 cases). Fifteen of 23 patients (65.2%) died of disease. Our results indicate that PLMS should be differentiated from ordinary leiomyosarcoma because of its high proliferative activities and rather aggressive biologic behavior. PMID- 11474288 TI - Evaluation of micrometastases in sentinel lymph nodes of cutaneous melanoma: higher diagnostic accuracy with Melan-A and MART-1 compared with S-100 protein and HMB-45. AB - Accurate diagnosis of micrometastases in sentinel lymph nodes of cutaneous melanoma is critical for proper clinical management. S-100 protein and HMB-45 are the traditional immunomarkers widely used for this purpose. However, the interpretation of micrometastases by these markers is difficult with significant reduction in the diagnostic accuracy. S-100 protein demonstrates immunoreactivity for other nonmelanoma cells and obscures nuclear details, which are crucial for the interpretation of single cell metastases. We compared the new melanoma markers, Melan-A (clone A103) and MART-1 (clone M2-7C10), with S-100 protein and HMB-45, by examining 77 formalin-fixed paraffin-embedded sections of sentinel lymph nodes from 13 cases of primary cutaneous melanoma. CD68 (PG-M1) and hematoxylin-eosin-stained sections were also studied. Four pathologists interpreted the staining pattern after concealing the identity of each immunomarker. Az values (area under receiver operating characteristic curve) with receiver operating characteristic curve were higher with Melan-A (0.9742) and MART-1 (0.9779) compared with S-100 protein (0.8034) and HMB-45 (0.8651), demonstrating a higher diagnostic accuracy with Melan-A and MART-1 with superior detection of melanoma micrometastases. Melan-A and MART-1 showed sharp cytoplasmic immunoreactivity, almost exclusively restricted to the melanoma cells. Therefore, Melan-A and MART-1 are recommended for the evaluation of micrometastases in sentinel lymph nodes of cutaneous melanoma as a routine alternative to S-100 protein and HMB-45. PMID- 11474289 TI - Familial fibrocystic pancreatic atrophy with endocrine cell hyperplasia and pancreatic carcinoma. AB - Understanding the pathology of familial pancreatic carcinoma may provide important insights into pancreatic tumorigenesis. We now describe in detail the pancreatic pathology of an autosomal dominant pancreatic carcinoma kindred with distinct clinical, genetic, and pathologic manifestations differing from all other reported forms of sporadic or familial pancreatic neoplasia. Affected individuals develop a prodrome of diabetes mellitus, pancreatic exocrine insufficiency, and characteristic pancreatic imaging abnormalities. Eleven family members have undergone total pancreatectomy, revealing a unique and characteristic fibrocystic, lobulocentric pancreatic atrophy. This was patchy to diffuse in distribution and was invariably associated with a nesidioblastosis like endocrine cell hyperplasia. All but one resected pancreas demonstrated glandular epithelial dysplasia: 10 had low-grade dysplasia (pancreatic intraductal neoplasia grade II of III or PanIN II) and seven also had high-grade dysplasia (pancreatic intraductal neoplasia grade III of III or PanIN III). Dysplasia was multifocal in small-to medium-sized duct-like structures within areas of acinar atrophy, microcystic change, and mucinous hyperplasia. Two pancreata had carcinomas of multiple and unusual histologic subtypes, including small cell undifferentiated carcinoma and giant cell anaplastic carcinoma. The findings in this kindred yield important information on a distinctive and previously unrecognized pancreatic cancer precursor. Recognition of this entity may help identify additional kindreds and perhaps the underlying genetic defect. As is the case for other familial cancers, the as yet unknown specific genetic defect may have wider implications for pancreatic cancer in general. PMID- 11474290 TI - p63, a p53 homologue, is a selective nuclear marker of myoepithelial cells of the human breast. AB - Myoepithelial cells (MCs) constitute the basal cell layer of normal mammary epithelia, and their identification is of particular diagnostic value because they are retained in most benign lesions while being lost in malignancy. Several MC immunocytochemical markers are currently available for diagnostic purposes, with special reference to smooth muscle-related antigens. p63 is a member of the p53 gene family, and its germline mutations are associated with severe mammary developmental defects in both rodents and humans. Different p63 isoforms have been identified, some of which (DeltaNp63) are preferentially expressed in the epithelial basal cells of different organs and have been considered as possible markers of stem cells/reserve cells. We investigated immunohistochemically 384 samples of normal and diseased human breast, including 300 invasive carcinomas, using four antibodies recognizing all p63 isoforms, or the DeltaNp63 isoforms. Twenty cytologic specimens were also investigated. Furthermore, snap-frozen tissue samples from three fibroadenomas and 10 invasive ductal carcinomas with their paired non-neoplastic tissues and three corresponding lymph node metastases were evaluated for the expression of p63 mRNA by RT-PCR. In normal breast tissue p63 immunoreactivity was confined to the nuclei of MCs. In all benign lesions p63 immunoreactive cells formed a continuous basal rim along the epithelial structures. Stromal cells, and in particular myofibroblasts, were consistently unreactive. Adenomyoepitheliomas showed nuclear staining in most neoplastic cells. A peripheral rim of p63-immunoreactive cells was retained surrounding lobular and ductal carcinoma in situ, although it was discontinuous as opposed to the normal structures. Invasive breast carcinomas were consistently devoid of nuclear p63 staining, with the exception of the two adenoid-cystic carcinomas, of the two ductal carcinomas with squamous metaplasia, and of 11 (4.6%) ductal carcinomas not otherwise specified, showing p63 immunoreactivity in a minor fraction (5-15%) of the neoplastic cells. In comparison with other MC markers, p63 was the most specific, being restricted exclusively to MCs, whereas antibodies to smooth muscle actin and, to a lesser extent, calponin also decorated stromal myofibroblasts. In the cytologic preparations p63 immunoreactivity was a consistent feature of "naked nuclei" and of a subset of cells surrounding benign epithelial clusters. RT-PCR experiments with primers specific for different p63 isoforms documented that normal tissues and fibroadenomas preferentially expressed the DeltaNp63 isoforms. Our study demonstrates that in normal and pathologic breast tissues MCs consistently express the DeltaNp63 isoforms. We suggest p63 as a reliable, highly specific, and sensitive MC marker in both histologic and cytologic preparations. Furthermore, because p63 immunoreactivity in adult epithelia is normally restricted to progenitor cells, it can be speculated that it might be a clue for the identification of the still elusive breast progenitor cells. PMID- 11474291 TI - Expression of Fli-1, a nuclear transcription factor, distinguishes vascular neoplasms from potential mimics. AB - Fli-1 protein, a member of the ETS family of DNAbinding transcription factors, is involved in cellular proliferation and tumorigenesis. Approximately 90% of Ewing's sarcoma/primitive neuroectodermal tumors (ES/PNET) have a specific translocation, t(11;22)(q24;q12), which results in fusion of EWS to Fli-1, and production of an EWS-Fli-1 fusion protein. We have recently shown that immunohistochemistry for the carboxy terminal of Fli-1 protein is sensitive and highly specific for the diagnosis of ES/PNET. In our earlier study we noted that among normal tissues only endothelial cells and small lymphocytes expressed Fli 1. Fli-1 expression in vascular neoplasms has not been previously studied. Formalin-fixed paraffin-embedded tissue from 54 vascular tumors and 75 nonvascular tumors were immunostained for Fli-1 (1:120, Sc 356, Santa Cruz Biotechnology, Santa Cruz, CA), after steam heat-induced epitope retrieval. Only cases with >10% of cells showing nuclear staining were accepted as positive. Cases without positive internal controls (endothelium and small lymphocytes) were not scored. Positive internal controls were present in 122 of 129 cases (95%). One vascular tumor (Kaposi's sarcoma) and 7 nonvascular tumors (2 epithelioid sarcomas and 5 carcinomas) without internal controls were not scored. Fli-1 was expressed by 50 of 53 vascular tumors scored (94%), including 20 of 22 angiosarcomas, 11 of 12 hemangioendotheliomas, 7 of 7 hemangiomas, and 12 of 12 Kaposi's sarcomas. In contrast, Fli-1 expression was absent in the 68 nonvascular tumors scored (0 of 68), including 16 sarcomas, 7 melanomas, and 45 carcinomas. The results of this study strongly suggest a role for Fli-1 as a novel marker of both benign and malignant vascular tumors. The sensitivity (94%) and specificity (100%) of Fli-1 with regards to the cases evaluated in this study equal or exceed those of the established vascular markers, CD31, CD34, and von Willebrand factor. As the first nuclear, rather than cytoplasmic or membranous marker of endothelium, Fli-1 immunostaining also generally lacks cytoplasmic staining artifacts that are the result of endogenous peroxidases or biotin. PMID- 11474292 TI - Colchicine toxicity: distinct morphologic findings in gastrointestinal biopsies. AB - Colchicine is an alkaloid with antimitotic ability used to treat a variety of medical conditions. Colchicine toxicity can result in multiorgan failure and death. The histopathologic features of colchicine toxicity in gastrointestinal biopsies have not been reported. Twenty-one gastrointestinal mucosal biopsies obtained from nine patients receiving oral colchicine therapy were studied. Immunohistochemical staining for Ki67 proliferation antigen was performed, and medical records of each patient were reviewed. All patients had a history of gout. Four patients with chronic renal failure also had clinical evidence of colchicine toxicity, and the other five patients did not. Distinct morphologic changes, seen as metaphase mitoses, epithelial pseudostratification, and loss of polarity, were seen in biopsy material from 4 of 4 (100%) patients with clinical colchicine toxicity. Three of these four cases (75%) also contained abundant crypt apoptotic bodies. These morphologic features were best seen in the biopsies from duodenum and gastric antrum, with relative sparing of the gastric body in the upper gastrointestinal tract. Ki67 staining demonstrated an expansion of the proliferating region in three available cases with clinical colchicine toxicity. These distinctive morphologic features were not seen in the five patients without clinical colchicine toxicity. These results indicate that colchicine toxicity can produce diagnostic morphologic features in gastrointestinal mucosal biopsies. Recognition of these features is important because colchicine toxicity can be fatal if undiagnosed clinically. PMID- 11474293 TI - Discriminatory immunohistochemical staining of urothelial carcinoma in situ and non-neoplastic urothelium: an analysis of cytokeratin 20, p53, and CD44 antigens. AB - Distinction of urothelial carcinoma in situ (CIS) from reactive atypia on the basis of morphology alone may be difficult in some cases. Because this distinction is therapeutically and prognostically critical, we attempted to determine if an immunohistochemical panel would help in this differential diagnosis. The immunoprofile of 21 cases of CIS and 25 non-neoplastic urothelia (15 urothelial biopsies with reactive atypia from patients without a history of bladder cancer and 10 normal ureter sections from nephrectomies performed for renal cell carcinoma) was determined using antibodies against cytokeratin 20 (CK20), p53, and CD44 (standard isoform). In the normal urothelium CK20 showed patchy cytoplasmic immunoreactivity in only the superficial umbrella cell layer and CD44 stained only the basal cells. Nuclear immunoreactivity to p53 varied from negative to weak and patchy. Reactive urothelium also showed CK20 immunoreactivity in only the umbrella cell layer in all 15 cases, and p53 nuclear staining was predominantly negative with occasional weak positivity in the basal and parabasal intermediate cells. CD44 was overexpressed in the entire reactive urothelium in 9 cases (60%) or focally positive in intermediate cells in 6 cases (40%). In contrast, CIS showed intense CK20 and p53 positivity (81% and 57%, respectively) in the majority (>50%) of malignant cells. CD44 staining revealed residual basal cells with membranous reactivity in 44% of the cases of CIS; however, the neoplastic cells were immunonegative in all cases. At least one positive immunomarker (CK20 or p53) was abnormally expressed in all cases of CIS. Abnormal expression of CK20 (increased), p53 (increased), and CD44 (decreased) in urothelial CIS, and increased expression of CD44 in reactive atypia allows more confident distinction of urothelial CIS from non-neoplastic urothelial atypias. From a differential diagnosis perspective, use of a panel of all three antibodies with morphologic correlation would be essential. PMID- 11474294 TI - Predicting cancer following a diagnosis of high-grade prostatic intraepithelial neoplasia on needle biopsy: data on men with more than one follow-up biopsy. AB - Most studies on the risk of cancer after high-grade prostatic intraepithelial neoplasia (PIN) on biopsy have been small (fewer than 100 men), have not analyzed in detail if histologic features can predict cancer, and have not assessed the risk of cancer with multiple repeat biopsies. We analyzed 245 men in whom the only abnormal finding on the initial biopsy was high-grade PIN and who had at least one follow-up biopsy. Repeat biopsy identified cancer in 32.2% of men. If only one follow-up biopsy had been performed on the 245 men, only 24.5% of men would have been found to have cancer. The only independent histologic predictor of a cancer diagnosis was the number of cores with high-grade PIN; risk of cancer: 30.2% with 1 or 2 cores, 40% with 3 cores, and 75% with >3 cores. The following did not predict cancer: number of high-grade PIN glands, maximum percentage of gland involved by high-grade PIN, nucleolar prominence, percentage of cells with prominent nucleoli, pattern of high-grade PIN (flat, tufting, micropapillary, cribriform), marked pleomorphism, digital rectal examination, transrectal ultrasound findings, family history of prostate cancer, serum prostate specific antigen (PSA) at time of high-grade PIN diagnosis, and rate of change of serum PSA. Eighty-one (33%) men had more than one follow-up biopsy; in these cases the following findings on the original high-grade PIN biopsy predicted cancer: the presence of mitoses, the number of positive cores, predominant micropapillary and cribriform high-grade PIN, and very large prominent nucleoli. Of 81 men with more than one follow-up biopsy, if the first repeat biopsy were benign, high-grade PIN, or atypical, the eventual cancer rate was 10%, 25.9%, and 57.1%, respectively (p = 0.002). Of 15 men with more than two repeat biopsies, only two (13.3%) had cancer. In summary, approximately one third of men with high-grade PIN on biopsy have cancer on follow-up. If cancer is not found on the first two follow-up biopsies, it will unlikely be found. Although clinical findings at the time of diagnosis of high-grade PIN are not useful to predict who might have cancer, histologic findings may help identify who needs additional biopsies. PMID- 11474295 TI - Thymoma with prominent cystic and hemorrhagic changes and areas of necrosis and infarction: a clinicopathologic study of 25 cases. AB - Twenty-five cases of thymoma with prominent cystic and hemorrhagic changes and areas of necrosis and infarction are presented. The patients were 11 women and 14 men between the ages of 18 and 73 years (median 45.5 years). Clinically, nine patients were asymptomatic and their mediastinal tumor was discovered on routine chest radiograph. Sixteen patients presented with symptoms of chest pain and cough. All patients underwent surgical resection of their tumor. Grossly, the tumors were described as well circumscribed and encapsulated, with the exception of two that showed infiltration of pleura and pericardium. The tumors measured from 4 to 13 cm in greatest dimension. On cut surface they showed prominent cystic areas and foci of hemorrhage and necrosis. Histologically, the tumors contained solid areas showing an admixture of round to oval epithelial cells devoid of atypia admixed with small lymphocytes in varying proportions. Cystic changes with areas of necrosis, infarction, and hemorrhage were present in all cases and comprised extensive areas of the tumors. The areas of infarction showed features of ischemic necrosis and were always intimately associated with vaso occlusive and thrombotic phenomena and with cystic and hyperplastic changes of adjacent thymic epithelium. Clinical follow-up in 14 patients showed that 11 were alive and well from 1 to 18 years after surgery (median follow-up 9 years). Three patients died: one of complications during the immediate postoperative period, one because of colonic adenocarcinoma 9 years after diagnosis of the mediastinal tumor, and one because of pneumonia 6 years later. The two patients with invasive tumors were lost to follow-up. The present study appears to indicate that areas of hemorrhage and necrosis in well encapsulated, noninvasive thymomas do not portend an adverse prognosis. PMID- 11474296 TI - Anatomic levels: important landmarks in penectomy specimens: a detailed anatomic and histologic study based on examination of 44 cases. AB - The majority of squamous cell carcinomas of the penis arise from the glans, and the prognosis is related significantly to the depth of invasion of crucial anatomic landmarks. Accurate information related to this can only be obtained when specimens are carefully evaluated grossly. Most pathologists in developed countries encounter resected specimens of penile carcinoma infrequently, and gross evaluation is occasionally suboptimal, potentially preventing obtaining reliable prognostic information. The four distinct levels of the glans penis are the epithelium, lamina propria, corpus spongiosum, and corpus cavernosum. A simple method for pathologic evaluation of the glans is presented. Noteworthy findings in our study of a South American population were that the distance from the lamina propria to tunica albuginea ranged from 7 to 13 to 6 mm at the dorsal, central, and ventral areas of the corpus spongiosum, respectively. The most distal portion of the corpus cavernosum was located within the glans in 34 of 44 cases and in the body of the penis in only 10. The corpus spongiosum was thinner in the former cases. These anatomic variations may bear on prognosis. PMID- 11474297 TI - Molecular genetic analysis of appendiceal mucinous adenomas in identical twins, including one with pseudomyxoma peritonei. AB - Pseudomyxoma peritonei (PMP) is a clinical syndrome characterized by mucinous ascites and peritoneal lesions composed of histologically bland to low-grade adenomatous mucinous epithelium within pools of extracellular mucin, often with an associated mucinous adenoma of the appendix. There is evidence that the peritoneal lesions in PMP are clonally derived from the associated appendiceal adenoma. Little is known about the molecular genetic alterations or hereditary factors involved in the development of appendiceal mucinous tumors and PMP. We report the only known example of appendiceal mucinous adenomas in identical twin brothers, one of whom developed PMP. We analyzed the status of the K-RAS and APC genes in these tumors using digital polymerase chain reaction and digital single nucleotide polymorphism (SNP) assay. Identical K-RAS mutations were detected in the appendiceal adenoma and peritoneal tumor from the twin with PMP, whereas the adenoma from the other twin harbored a different mutation. Digital SNP analysis demonstrated loss of heterozygosity of APC only in the adenoma from the twin without PMP but not from the appendiceal or peritoneal tumors of the twin with PMP. The adjacent normal tissue in each case retained both APC alleles. The K-RAS mutational analysis supports the view that PMP is clonally derived from the associated appendiceal mucinous adenoma. The lack of loss of heterozygosity of APC in the adenoma and peritoneal tumor from the twin with PMP suggests that loss of heterozygosity of APC is not necessarily involved in the development of all appendiceal adenomas or PMP. The different types of mutations in K-RAS and the different allelic status of the APC locus in the tumors from both twins suggest that mutation in K-RAS and loss of heterozygosity of APC occurs somatically in adenomas and is independent of the identical genetic background of the twins. PMID- 11474298 TI - Diagnostic 'individuality'. PMID- 11474299 TI - 'Individuality' in the specialty of surgical pathology. PMID- 11474300 TI - On variability, standardization, and error in diagnostic pathology. PMID- 11474301 TI - 'Individuality' in surgical pathology. PMID- 11474302 TI - Sarcomas arising in dermatofibrosarcoma protuberans: collision or illusion? PMID- 11474304 TI - Peptide growth factors in the intestine. AB - A continuously increasing number of regulatory peptides has been demonstrated to be expressed in the intestine and to modulate several functional properties of various intestinal cell populations, including the intestinal epithelium and lamina propria cell populations. These regulatory peptides include members of the epidermal growth factor (EGF) family, the transforming growth factor beta (TGF beta) family, the insulin-like growth factor (IGF) family, the fibroblast growth factor (FGF) family, the trefoil factor (TFF) family, the colony-stimulating factor (CSF) family, and a few other seemingly unrelated regulatory peptides, such as hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and various interleukins, interferons and tumour necrosis factor-related proteins. In addition to the well-known effects on cell proliferation, these regulatory peptide factors regulate several other functional properties of epithelial and other cell populations, such as differentiation, migration, and extracellular matrix deposition and degradation. This review is designed not to discuss all the identified factors in detail but to highlight some of the basic principles of growth factor action in the intestine. It focuses mainly on classical growth factors rather than interleukins and interferons. PMID- 11474305 TI - Defensins -- innate immunity at the epithelial frontier. AB - Physical barrier function was formerly believed to play the major role in mucosal protection against luminal bacteria. This view has now been challenged by the discovery of specialized molecules that possess antimicrobial activity. More than 100 peptides have been identified so far, and the number is still growing. These peptides are distributed widely and conserved throughout phylogeny. The epithelial expression of antimicrobial peptides is of particular interest as many pathogens adhere to epithelial surfaces and may eventually invade the host. This rapidly acting defence system of innate immunity is already engaged before adoptive immune interactions take place. These antimicrobial peptides consist of constitutive and inducible forms, potentiating this barrier function in terms of an inflammatory response. One important subgroup of antimicrobial peptides is the family of defensins, which are classified as alpha (alpha-) and beta (beta-) defensins. Eight different peptides with varying antimicrobial properties have been identified. They are distributed widely in humans, and organ-specific expression patterns have been observed. Homologous peptides have been found in other mammals, vertebrates, invertebrates, insects and plants. The identification of alpha-defensins and their murine counterparts, cryptdins, in the small intestine prompted intensive research into epithelial antimicrobial defence. PMID- 11474306 TI - Cytokines in the liver. AB - Cytokines comprise a group of small proteins released from cells in order to influence the function of other cells. By binding to highly specific cell-surface receptors, they trigger a vast array of intracellular signalling cascades. Cytokines have been described as interleukins, growth factors, interferons and chemokines. Unlike hormones, which act in a similar way, cytokines are produced by many different types of cell and act on many other types. Most of them are produced only after certain stimuli. The most intense field of cytokine activity is without doubt host defence. The liver resembles a central organ of cytokine activity due to the fact that it hosts hepatocytes, which are highly susceptible to the activity of cytokines in a variety of physiological and pathophysiological processes. Moreover, the non-parenchymal cells of the liver, in particular Kupffer cells (KCs), the resident tissue macrophages of the liver, are able to synthesize a variety of cytokines that may act systemically on any other organ of the body, or in a paracrine manner on hepatocytes and other non-parenchymal liver cells. A classic example of how cytokines act can be observed during the acute phase reaction discussed in this article. The role of cytokines in liver development, acute liver injury, liver regeneration, liver fibrosis and liver metastasis is also discussed. PMID- 11474307 TI - Apoptosis in liver disease. AB - A variety of biological functions are regulated through extracellular signals. Amongst the best studied examples is growth control, which is achieved by the regulatory function of growth factors. In recent years it has become apparent that cell death (apoptosis) is controlled in a similar fashion.Apoptosis, firstly a morphologically defined process, is a highly controlled type of cell death that plays a critical role in embryonic development, deletion of autoreactive T-cells and adult tissue homoeostasis. There is increasing evidence that derangement of the apoptotic program is the underlying cause of a series of diseases including liver diseases. The deadly program can be initiated by ligand binding to membrane bound receptors such as CD95 (Fas), which is the most prominent cell death inducing member of the TNF receptor superfamily. The core of the subsequently activated intracellular machinery is formed by a set of proteases, namely caspases. Once activated, they orchestrate the complete destruction of the cellular skeleton leading to the typical apoptotic morphology. This review focuses on the underlying mechanism leading to derangement of the usually highly controlled apoptotic program in different liver diseases. PMID- 11474308 TI - The role of the liver in the production of thrombopoietin compared with erythropoietin. AB - The liver plays an important role in the production of haemopoietic hormones. It acts as the primary site of synthesis of erythropoietin (EPO) in the fetal stage, and it is the predominant thrombopoietin (TPO)-producing organ for life. In contrast to that of EPO and other liver proteins, the hepatic synthesis of TPO is influenced little by external signals. Hepatocytes express the TPO gene in a constitutive way, i.e. irrespective of the level of platelets in blood. Megakaryocytes and platelets remove the hormone from blood by means of their high affinity TPO receptors. Normally, the plasma level of TPO is relatively low ( approximately 10(-12) mol/l). However, in thrombocytopenic states due to marrow failure or bleeding, the concentration of circulating TPO may increase greatly. The simple feedback regulation by TPO and its target cells is efficient in maintaining constant platelet numbers in healthy people. Persisting thrombocytopenia develops only in severe liver or marrow failure. On the other hand, an increase in circulating TPO and interleukin 6 (IL-6) may cause reactive thrombocytosis in inflammatory diseases, including cancer. The indications for recombinant human thrombopoietin (rHuTPO) therapy and its impact on transfusion medicine are still under investigation. PMID- 11474309 TI - The (13)C-urea breath test for non-invasive diagnosis of Helicobacter pylori infection: which procedure and which measuring equipment? AB - 1) The (13)C-urea breath test is one of the most important non-invasive methods for detecting Helicobacter pylori infection in both pre- and post-treatment phases. 2) In order to render the test less expensive and more rapid, a series of modifications have recently been proposed concerning the dose of (13)C-urea and its formulation, as well as the type of measuring equipment used for detecting (13)CO(2) in breath samples. 3) The use of small doses (40--50 mg) of isotope in gelatin capsules or mixed with citric acid in rapid-releasing tablets seems to be promising as it shortens sampling time to 10--20 min. 4) Infrared spectroscopy and laser-assisted ratio analysis equipment are two new valid and less expensive alternatives to conventional isotope mass spectrometry for (13)C analysis in breath. The (13)C-urea breath test is one of the most accurate non-invasive tests for diagnosing Helicobacter pylori infection. This method has become increasingly popular in the last decade since it employs an innocuous, non-radioactive isotope that can be safely used in both children and adults. The test has been recently simplified by reducing the dose of (13)C-urea used, by adopting shorter sampling times and by using new and cheaper measuring methods for (13)C analysis in the breath samples. These include nondispersive isotope-selective infrared spectroscopy (NDIRS) and laser-assisted ratio analysis (LARA), which greatly differ from each other in their technical characteristics and cost. Both NDIRS and LARA are valid alternatives to conventional isotope ratio mass spectrometry (IRMS), although LARA appears to be more suitable for large laboratories or gastroenterological centres, which need to process many breath samples per day, whereas NDIRS seems to be more suitable for small laboratories or doctors' offices. PMID- 11474310 TI - Qualitative and quantitative clinical evaluation of the laser-assisted ratio analyser for detection of Helicobacter pylori infection by (13)C-urea breath tests. AB - OBJECTIVES: Currently, the (13)C-urea breath test is the reference method for non invasive diagnosis of Helicobacter pylori infection and therapy control. Therefore, new technologies have been developed to measure the ratio of (13)CO(2)/(12)CO(2) in breath. The laser-assisted ratio analyser (LARA) device is based on optogalvanic effects of the stimulated CO(2) molecules. DESIGN: In this study, the LARA system is prospectively compared to conventional isotope ratio mass spectrometry (IRMS) analysis of (13)C-urea breath tests. METHODS: The (13)C urea breath test was used to screen 103 patients for H. pylori infection. Breath samples were analysed by LARA and IRMS techniques. RESULTS: Seven breath tests could not be analysed by the LARA system, one by IRMS. Out of the remaining 95 breath tests, 13 were positive for H. pylori infection (13.7%). In reference to IRMS analysis (with a cut-off of > 5 delta per thousand at 30 min), LARA produced one false positive and one false negative breath test result giving a sensitivity of 92.3% and a specificity of 98.8%. The mean difference in delta over baseline values between IRMS and LARA measurements was 2.02 delta per thousand +/- 5.48 delta per thousand. CONCLUSION: LARA allows the reliable qualitative evaluation of 13C-urea breath tests, but the quantitative results differ from IRMS findings. PMID- 11474311 TI - Relapse prevention in reflux oesophagitis with regard to Helicobacter pylori status: a double-blind, randomized, multicentre trial to compare the efficacy of pantoprazole versus ranitidine. AB - OBJECTIVE: To compare prospectively the effectiveness of 1 year's treatment with pantoprazole versus ranitidine in order to prevent relapse after initial cure of reflux oesophagitis. For the first time the influence of the initial Helicobacter pylori status on therapeutic results was also taken into account. METHODS: In order to cure reflux oesophagitis, 396 patients with Savary/Miller stage II or III reflux oesophagitis were treated with pantoprazole 40 mg once daily for 8 weeks. Those who were H. pylori positive (n = 140) were also given 1 week of eradication treatment with clarithromycin 2 x 250 mg daily, metronidazole 2 x 400 mg daily, and a further 40 mg pantoprazole daily. The 303 patients who were endoscopically cured after the 8-week period were randomized and treated with either pantoprazole 20 mg (n = 199) or ranitidine 150 mg (n = 104) daily in double-blind fashion. The primary objective was to assess the time to endoscopically proven recurrence of reflux oesophagitis. RESULTS: In the intention-to-treat (ITT) population, 66.3% (118/178) of the pantoprazole group and 34.0% (32/94) of the ranitidine group showed neither endoscopic nor clinical symptoms of relapse after the 1-year treatment period (P < 0.0001) (per-protocol populations: 70.3% [109/155] in the pantoprazole group and 39.4% [28/71] in the ranitidine group). In the pantoprazole group, the relapse rate in initially H. pylori-positive patients who underwent eradication was 30.9% (17/55) and in H. pylori-negative patients 29% (29/100). CONCLUSIONS: Long-term treatment with 20 mg pantoprazole daily to prevent relapse of reflux oesophagitis in H. pylori negative patients is significantly more effective than 150 mg ranitidine daily. The initial H. pylori eradication treatment does not influence the outcome of the long-term treatment. PMID- 11474312 TI - Percutaneous endoscopic gastrostomy: complications and suggestions to avoid them. AB - OBJECTIVES: Percutaneous endoscopic gastrostomy (PEG) tubes have become an excellent alternative for the long-term management of patients with proximal obstructions of the gastrointestinal tract. However, their use has limitations and can be associated with serious complications. We therefore studied the frequency and severity of complications related to the use of PEG tubes in our clinic. DESIGN: All adults (aged 18 years and above) in whom a PEG tube was placed between January 1 1994 and January 1 1999 at the Free University Hospital in Amsterdam were included in this study. In initial cases, the indication and procedure were individually judged according to a liberal protocol. However, after several major complications, a strict procedure protocol was implemented in September 1996. RESULTS: During the study period, 263 PEG tubes were placed in 254 patients with head and neck cancer (n = 183; 70%), neurological disorders (n = 52; 20%) or severe upper gastrointestinal motility disorders (n = 28; 11%). In period I, 167 PEG tubes were placed and in period II, 96 PEG tubes were inserted. Patients were followed for a median 111 days. Minor complications occurred in 13% of the patients. Major complications occurred in 8% of the patients. In period I, the percentage of major complications was higher than in period II (9.5% versus 6%). CONCLUSION: PEG tube placement is a safe procedure when performed according to strict guidelines. By doing so, PEG tubes allow optimal feeding for prolonged periods with the occasional need for replacement of the tube. PEG tubes should not be introduced in acutely ill patients, patients with a short life expectancy and preferably not to patients with severe coughing. PMID- 11474313 TI - Gastric myoelectrical activity and gut hormone secretion in myotonic dystrophy. AB - BACKGROUND: Myotonic dystrophy (MD) is a systemic disease affecting striated, cardiac and smooth muscles, as well as nerve structures and endocrine glands. Patients with MD may suffer from slow gastric emptying. OBJECTIVE: To study electrogastrograms (EGG) and postprandial gut hormone profiles in MD in order to evaluate whether disturbances in these regulatory mechanisms could explain, or contribute to, the delayed gastric emptying. SUBJECTS: Ten patients with MD complaining of symptoms consistent with slow gastric emptying, and ten healthy matched controls. METHODS: After an overnight fast, the patients and the control subjects were examined with standard EGG using surface electrodes before and during intake of a standard meal. Blood tests were drawn at regular time intervals for hormone analyses. RESULTS: The EGG in MD showed a reduced amount of normal three cycles per minute activity compared with controls (P < 0.04). The dominant frequency in MD was less stable than in controls (P < 0.03), and the power of the signal showed less increase after a meal. The postprandial increase in plasma motilin (P < 0.05) and glucagon-like peptide-1 (GLP-1) (P < 0.001) was significantly less pronounced in MD compared with controls, whereas the plasma concentrations of cholecystokinin (CCK), neurotensin (NT), peptide YY (PYY) and somatostatin (SOM) did not differ significantly. CONCLUSION: Disturbed electrophysiological control of the stomach and impaired secretion of gastrointestinal peptide hormones could contribute to slow gastric emptying in MD. Combined impairment of gastric pacing and gastrointestinal hormone responses was found in patients with the most prominent retardation of gastric emptying. PMID- 11474314 TI - The effects on gastroduodenal mucosa of a new nonsteroidal anti-inflammatory drug, amtolmetin-guacyl, versus piroxicam in healthy volunteers: a short-term, double-blind, endoscopically controlled study. AB - AIM: Amtolmetin-guacyl (AMG) (2-[2[1-methyl-5-(4-methylbenzoyl) pyrrol-2-yl] acetamido] acetic acid 2-methoxyphenyl ester) is a recent drug that, in preliminary studies, has shown effective anti-inflammatory properties with improved gastrointestinal safety. Our study was designed to investigate the effects of AMG and piroxicam on gastroduodenal mucosa in healthy volunteers. MATERIALS AND METHODS: Forty-two healthy volunteers aged 18--45 years were randomized in a double-blind manner to AMG 1200 mg for 2 days and 600 mg for 12 days, or piroxicam 40 mg for 2 days and 20 mg for 12 days. Endoscopic evaluation and laboratory tests were performed at baseline and at the end of the treatment. The mucosa was evaluated by endoscopy using a predefined scale: the score could range from 0 to 4. Only volunteers with endoscopy grade 0-1 entered the trial. RESULTS: The median post-treatment endoscopy gastric injury scores were 1 (range 0--4) in the AMG-treated volunteers and 3 (range 0--4) in the piroxicam-treated volunteers (P = 0.04). There were two cases with an endoscopic gastric score of 4 in the AMG group, and seven in the piroxicam group (P = 0.1). The corresponding values in the duodenum were 1/21 volunteers in the AMG group and 1/21 in the piroxicam group. Eight out of 11 subjects with an endoscopic score of 4 were Helicobacter pylori negative, and 3/11 were infected by the micro-organism. Different adverse reactions were reported by 15/21 volunteers (71%) in the AMG group and by 12/21 (57%) in the piroxicam group. None of these events resulted in interruption of the study. CONCLUSIONS: AMG is a new anti-inflammatory drug with limited gastric toxicity. If these findings are confirmed on a wider scale in long-term trials, then the drug might become a valid alternative to current treatments, especially for patients such as those with rheumatoid arthritis who need steroids and second-line drugs simultaneously. PMID- 11474315 TI - Characterization of antigen-presenting dendritic cells in the peripheral blood and colonic mucosa of patients with ulcerative colitis. AB - OBJECTIVE: Increased lymphocyte activation and production of inflammatory cytokines are implicated in the pathogenesis of ulcerative colitis. Because antigen-presenting dendritic cells play a cardinal role in the activation and survival of activated lymphocytes, the aim of the present study was to characterize dendritic cells in ulcerative colitis. DESIGN: This study was designed to compare the phenotypes and functions of peripheral blood dendritic cells among healthy normal volunteers and patients with ulcerative colitis or Crohn's disease. Activated dendritic cells were also localized at the colonic mucosa. METHODS: Peripheral blood dendritic cells were generated from 15 patients with ulcerative colitis, 10 patients with Crohn's disease and 15 healthy control volunteers. The stimulatory capacities of dendritic cells were analysed in an allogenic mixed lymphocyte reaction. Nitric oxide was detected by the Griess method. Single- and dual-colour flow cytometry was employed to study the levels of maturation of dendritic cells. Activated dendritic cells were localized immunohistochemically in the colonic mucosa. RESULTS: In comparison to normal controls, peripheral blood dendritic cells from patients with ulcerative colitis showed significantly increased stimulatory capacities (P < 0.05) and produced significantly higher levels of nitric oxide (P < 0.05). The numbers of activated dendritic cells were also significantly higher in ulcerative colitis (P < 0.05). Mature and activated dendritic cells expressing the CD83 antigen were detected at the inflamed colonic mucosa in patients with ulcerative colitis and Crohn's disease. CONCLUSIONS: Activated and mature dendritic cells may have a role in the induction of an exacerbated immune response in ulcerative colitis. This study provides the scientific and logical basis for blocking the maturation and activation of dendritic cells in ulcerative colitis as a new therapeutic intervention. PMID- 11474316 TI - Expression of insulin-like growth factor binding proteins and collagen in experimental colitis in rats. AB - OBJECTIVES AND DESIGN: Crohn's disease is complicated by smooth muscle hyperplasia and stricture formation. Insulin-like growth factor (IGF)-1 and insulin-like growth factor binding proteins (IGFBPs) may be involved in stimulating intestinal smooth muscle growth and collagen synthesis. Therefore, we investigated the expression of IGFBPs, collagen and collagenase activity in rat colitis and the effects of IGF-1 on IGFBP and collagen expression in rat colonic smooth muscle cells. METHODS: Animals were sacrificed during a 4-week time course of 2,4,6-trinitrobenzenesulphonic acid (TNBS)-induced colitis. RNA from the animals' colons was blotted and hybridized with collagen-1 and IGFBP mRNA probes. Tissue proteins were screened for IGFBPs by Western ligand blotting. Collagenase activity was measured by zymography. Rat colonic smooth muscle cells in primary culture were incubated with IGF-1 then collagen-1, and IGFBP mRNAs and proteins were measured. RESULTS: In the rat tissue, IGFBP-3 mRNA and protein were increased 2 h after induction of colitis. IGFBP-4 mRNA was elevated after 2 h and IGFBP-4 protein after 4 h. IGFBP-5 mRNA was upregulated after 2 h with a peak at 12 h. IGFBP-5 protein was upregulated after 1 h and reached a peak at 3 days. Collagen-1 mRNA was increased after 5 days. Collagenase levels were decreased after 1 h and returned to normal by 28 days. In rat colonic smooth muscle cells, IGF-1 increased collagen-1 and IGFBP-5 expression. CONCLUSION: We demonstrated an upregulation of IGFBP and collagen expression and a downregulation of collagenase in rat colitis. In colonic smooth muscle cells, we found an upregulation of collagen-1 and IGFBP-5 following IGF-1 incubation. These results suggest an important role of IGF-1 in the collagen synthesis in colitis, mediated by IGFBPs. PMID- 11474317 TI - Interferon alpha therapy in haemophilic patients with chronic hepatitis C: a French multicentre pilot study of 58 patients. AB - BACKGROUND AND OBJECTIVES: Information about the long-term efficacy of interferon alpha (interferon-alpha) in haemophilic patients with chronic hepatitis not co infected with the human immunodeficiency virus (HIV-1) is still limited. Previous studies seemed to indicate a low rate of response. The aim of this study was to evaluate the safety and long-term efficacy of interferon treatment in multi transfused haemophiliacs. METHODS: Fifty-eight haemophiliacs were scheduled to receive 3 MU of interferon-alpha 2b three times a week for 12 months. The patients were followed up for at least 24 months post-treatment. Response was assessed by measurements of serum hepatitis C virus (HCV) RNA. RESULTS: Twenty four patients (41.4%) dropped out. Except for seven patients, the symptoms that led to interrupting interferon treatment would probably not have resulted in the same decision in non-haemophilic patients. One patient developed an inhibitor to the deficient clotting factor without haemorrhagic consequences. In an intent to treat, the sustained virological response rate was 14%. However, when considering only the 34 patients who received the full treatment, HCV-RNA was cleared in eight patients (23%). CONCLUSIONS: This study suggests that multi-transfused haemophiliacs with chronic hepatitis not co-infected with HIV-1 respond to prolonged treatment with interferon-alpha in a similar proportion to that observed in non-haemophiliacs. There was a high rate of patients who did not complete the interferon-alpha treatment, and this seems to be characteristic of this patient population. PMID- 11474318 TI - Sister Joseph's nodule in hepatocellular carcinoma. AB - We describe three black South African patients in whom hepatocellular carcinoma metastasized to the umbilicus. Sister Joseph's nodule has previously been reported in only two patients with this tumour. A number of routes for this spread are possible: malignant hepatocytes in the portal venous system may reach the umbilicus via a patent umbilical vein; the tumour may propagate directly along the ligamentum teres hepatis to the umbilicus; contiguous spread of the tumour to the umbilicus from anterior peritoneal tissue, either directly infiltrated by hepatocellular carcinoma or the site of metastatic nodules, may occur; embolization of malignant hepatocytes to the umbilicus might take place by way of its arterial blood supply; or hepatocellular carcinoma might reach the umbilicus as a result of retrograde lymphatic spread from para-aortic lymph nodes or from the anterior abdominal wall, to which the tumour has metastasized. PMID- 11474319 TI - Oropharyngeal pemphigus in a patient with chronic hepatitis C during interferon alpha-2a therapy. AB - There are a few reports in the literature concerning pemphigus induced by interferon given for hepatitis C. We present the case of a 28-year-old woman with post-transfusional chronic hepatitis C who developed ulcers and vesicles on her tongue, cheeks, posterior oropharynx and vocal cords 5 months after beginning treatment with recombinant interferon alpha-2a. The direct and indirect immunofluorescence was diagnostic of pemphigus vulgaris. The drug was promptly withdrawn; the patient was medicated with prednisolone and azathioprine and recovered only 3 months later. Although there are several publications describing the occurrence of other autoimmune diseases in patients receiving interferon alpha therapy, this is the first report of a pemphigus induced by interferon in hepatitis C patients involving oropharyngeal and laryngeal mucosae without cutaneous involvement. PMID- 11474320 TI - Ectopic hepatocellular carcinoma arising in the left chest wall: a long-term follow-up. AB - We report the case of a 66-year-old man with chronic hepatitis C and a slowly growing left chest wall mass. Two years after the patient first noticed the mass, it was resected. A diagnosis of hepatocellular carcinoma (HCC) was established. The liver was studied by ultrasound, computed tomography (CT), magnetic resonance imaging (MRI) and angiography, but no mass was found. Blind liver biopsy showed mild chronic hepatitis without cirrhosis or HCC. Three years after the discovery of the chest wall HCC, no liver mass had appeared at CT and MRI. We conclude that solitary extrahepatic HCC (i) may arise in ectopic liver tissue; (ii) should not be considered as a metastasis of an occult HCC; and (iii) can be amenable to cure through resection. PMID- 11474321 TI - Haemobilia causing acute pancreatitis after percutaneous liver biopsy: diagnosis by magnetic resonance cholangiopancreatography. AB - Magnetic resonance cholangiopancreatography (MRCP) has received much attention as a non-invasive alternative to endoscopic retrograde cholangiopancreatography, primarily for investigation of choledocholithiasis, but also for evaluation of less common biliary anomalies. We present a case of haemobilia causing acute pancreatitis after percutaneous liver biopsy in which the diagnosis could be made clearly by MRCP, thus avoiding endoscopic retrograde cholangiopancreatography and sphincterotomy. PMID- 11474322 TI - Insulin action and long-term electrically induced training in individuals with spinal cord injuries. AB - PURPOSE: Individuals with spinal cord injuries (SCI) have an increased prevalence of insulin resistance and type 2 diabetes mellitus. In able-bodied individuals, training with large muscle groups increases insulin sensitivity and may prevent type 2 diabetes mellitus. However, individuals with SCI cannot voluntarily recruit major muscle groups, but by functional electrical stimulation (FES) they can now perform ergometer bicycle training. METHODS: Ten subjects with SCI (35 +/ 2 yr (mean +/- SE), 73 +/- 5 kg, level of lesion C6--Th4, time since injury: 12 +/- 2 yr) performed 1 yr of FES cycling (30 min x d(-1), 3 d x wk(-1) (intensive training)). Seven subjects continued 6 months with reduced training (1 d x wk(-1) (reduced training)). A sequential, hyperinsulinemic (50 mU x min(-1) x m(-2) (step 1) and 480 mU x min(-1) x m(-2) (step 2)), euglycemic clamp, an oral glucose tolerance test (OGTT), and determination of GLUT 4 transporter protein in muscle biopsies were performed before and after training. RESULTS: Insulin stimulated glucose uptake rates increased after intensive training (from 4.9 +/- 0.5 mg x min(-1) x kg(-1) to 6.2 +/- 0.6 mg x min(-1) x kg(-1) (P < 0.008) (step 1) and from 9.0 +/- 0.8 mg x min(-1) x kg(-1) to 10.6 +/- 0.8 mg x min(-1) x kg( 1) (P = 0.103) (step 2)). With the reduction in training, insulin sensitivity decreased to a similar level as before training (P > 0.05). GLUT 4 increased by 105% after intense training and decreased again with the training reduction. The subjects had impaired glucose tolerance before and after training, and neither glucose tolerance nor insulin responses to OGTT were significantly altered by training. CONCLUSIONS: Electrically induced bicycle training, performed three times per week increases insulin sensitivity and GLUT 4 content in skeletal muscle in subjects with SCI. A reduction in training to once per week is not sufficient to maintain these effects. FES training may have a role in the prevention of the insulin resistance syndrome in persons with SCI. PMID- 11474323 TI - Knee bracing after ACL reconstruction: effects on postural control and proprioception. AB - PURPOSE: To evaluate the effects an anterior cruciate ligament (ACL) brace has on various measures of knee proprioception and postural control. METHODS: Thirty subjects (mean age 27 +/- 11 yr) having undergone unilateral ACL reconstruction were tested with and without wearing their own custom-fit brace on their involved limb. Proprioception was assessed using joint angle replication tests completed on an isokinetic dynamometer. Postural control was assessed using a series of single-limb standing balance tests completed on a force platform. The balance tests included: 1) standing on the stable platform with eyes open, 2) standing on a foam mat placed over the platform with eyes open, 3) standing on the platform with eyes closed, and 4) standing on the platform after landing from a maximal single-limb forward hop. RESULTS: The brace provided a small but statistically significant improvement in proprioception (mean reduction in error scores between target and reproduced angles = 0.64 +/- 1.4 degrees, P = 0.02). For the postural control tests, there was a significant brace condition by test situation interaction (P = 0.02), with the brace providing a small but statistically significant improvement during the test completed on the stable platform with eyes open (mean reduction in center of pressure path length = 4.2 +/- 8.4 cm, P = 0.02) but not during the other more challenging test situations. Additional post hoc analyses indicated that the relationship between knee proprioception and postural control measures were low and not significant (r = 0.003 to 0.19, P > 0.32), consistent with the suggestion that changes in knee proprioception can occur in the absence of substantial changes in postural control. Also, standing balance tests that challenged the somatosensory contribution to postural control (i.e., those completed on foam, or with eyes closed) were significantly related to single-limb forward hop distances (r = -0.4, P < 0.05), whereas performance during the proprioception test was not (r = 0.1, P > 0.50). CONCLUSIONS: In general, bracing appears to improve performance during tasks characterized by relatively limited somatosensory input but not during tasks characterized by increased somatosenory input. The small magnitude of the improvements, coupled with their apparent lack of carry over to more difficult and functionally relevant tasks, questions the clinical benefit of the present effects of bracing. PMID- 11474324 TI - Prior meal enhances the plasma glucose lowering effect of exercise in type 2 diabetes. AB - PURPOSE: To compare the changes in plasma glucose and insulin levels in response to 1 h of exercise performed at 60% of VO(2peak) either in the fasted state or 2 h after a standardized breakfast in subjects with type 2 diabetes. METHODS: Ten sedentary men with type 2 diabetes treated with oral agents and not under strict metabolic control were tested on two occasions (fasted and fed state) in a random order at a 1-wk interval. RESULTS: Plasma glucose was slightly but not significantly higher at the beginning of exercise performed in the fed state versus the fasted state (12.4 +/- 1.3 vs 11.1 +/- 1.1 mmol x L(-1) respectively; mean +/- SE, P = 0.06). However, after exercise, plasma glucose levels were much lower in the fed state (7.6 +/- 1.1 mmol x L(-1)) compared with the fasted state (10.0 +/- 1.0 mmol x L(-1); P = 0.009). Insulin levels were higher at the beginning of the exercise bout performed in the fed state (177 +/- 26 vs 108 +/- 19 pmol x L(-1); P < 0.05) and during exercise. Similar respiratory exchange ratio at identical workload indicated that the difference in glycemic response was not due to differences in whole body substrate utilization. Plasma concentrations of free fatty acids, glucagon, epinephrine, and norepinephrine were also similar during both experiments. CONCLUSIONS: One hour of aerobic exercise has a minimal impact on plasma glucose level when performed in fasted moderately hyperglycemic men with type 2 diabetes but induces an important decrease in plasma glucose level when performed 2 h after breakfast. Because glucose utilization increased similarly during exercise in both conditions, the higher insulin levels after the meal might have blunted glucose production, creating an imbalance between total glucose production and total peripheral utilization in the fed state in contrast to the fasted state. PMID- 11474325 TI - Is there a disassociation of maximal oxygen consumption and maximal cardiac output? AB - PURPOSE: The purpose of the present study was to determine whether maximal cardiac output (Q) is affected by the duration of the maximal exercise test. METHODS: Nine healthy men (N = 6) and women (N = 3) performed three separate maximal treadmill exercise tests, separated by at least 24 h, and underwent a body composition assessment by hydrostatic weighing, all within a 2-wk period. A baseline maximal exercise test was performed to establish VO(2max). The second and third tests, assigned randomly, were designed to elicit the subjects' predetermined VO(2max) in either 6 or 12 min. Heart rate (HR), blood pressure (BP), minutes of ventilation, and oxygen consumption (VO(2)) were measured during all tests. At the end of the 6- and 12-min tests, Q was measured using an acetylene rebreathing technique. Stroke volume (SV), mean arterial pressure (MAP), total peripheral resistance (TPR), and arteriovenous O(2) difference were calculated using standard equations. RESULTS: Repeated-measures ANOVA indicated that there were no significant differences in HR and VO(2max) between the baseline, 6-min, and 12-min tests. Paired t-tests revealed significantly greater Q (25.1 +/- 5.6 vs 23.7 +/- 5.2 L.min-1) and SV (138.3 +/- 31.5 vs 130.5 +/- 31.2 mL) in the 6- versus 12-min tests, respectively. There were no significant differences in systolic BP, diastolic BP, MAP, TPR, or arteriovenous O(2) difference. CONCLUSIONS: Despite there being no difference in VO(2max) between the two tests, the 6-min maximal exercise test resulted in a significantly greater Q than the 12-min test, because of a significantly greater SV. Thus, there was a disassociation between VO(2) and Q during maximal exercise. PMID- 11474326 TI - Effect of intermittent high-intensity exercise on gastric emptying in man. AB - PURPOSE: The effect on gastric emptying of brief intermittent high-intensity sprints and a moderate-intensity steady-state cycle exercise was studied. METHODS: Eight healthy male subjects were each studied at rest (R), during steady state exercise at a constant power output corresponding to 66% of their VO(2max) (C66), during intermittent high-intensity exercise at a power output averaging 66% of their VO(2max) (I66), and during intermittent high-intensity exercise at a power output averaging 75% of their VO(2max) (I75). Gastric emptying was measured using the double-sampling gastric aspiration technique. Subjects ingested 600 mL of a 6% carbohydrate-electrolyte solution immediately before exercise or seated rest. RESULTS: The volume of test solution in the stomach was less at all time points on trial I66 than on trial I75 (P = 0.023). The rate of gastric emptying, expressed as the median (range) time (minutes) taken to empty half the test meal volume (t(1/2)), was not different on trials R (20 (7--30)) and C66 (21 (7--49)), and was faster than on trial I75 (62 (27--100); P = 0.003 and P = 0.005, respectively). Median t(1/2) was faster on trial R than on trial I66 (30 (15--74) min; P = 0.019), but no difference was detected between C66 and I66 or between I66 and I75. However, over the initial 30 min period after ingestion, the median (range) volume of test drink delivered to the duodenum was faster (P < 0.01) on trials R (387 (296--541) mL) and C66 (389 (165--584) mL) than on trials I66 (331 (191--494) mL) or I75 (249 (79-335) mL). CONCLUSION: The data demonstrate that gastric emptying of liquids is slowed during brief intermittent high-intensity exercise compared with rest or steady-state moderate exercise. PMID- 11474327 TI - Does endurance training affect orthostatic responses in healthy elderly men? AB - PURPOSE: To investigate the effects and time course of endurance training on the regulation of heart rate (HR), arterial pressure (AP), norepinephrine (NE), and plasma volume (PV) during orthostatic stress in healthy elderly men. METHODS: Thirty-one healthy men (65--75 yr) were randomly allocated into endurance training (N = 20, EX) and control (N = 11, CON) groups. The EX group cycled 3 d x wk(-1) for 30 min at 70% VO(2peak) for 12 wk x VO(2peak) was determined on an electronically braked cycle ergometer, before training and after 4, 8, and 12 wk of endurance training. The immediate (initial 30 s), early steady-state (1 min), and prolonged (5, 10, 15 min) beat-by-beat HR and AP responses during 90 degrees head-up tilt (HUT) were measured at least 3 d after each VO(2peak) test. Spontaneous baroreflex slopes were determined by application of linear regression to sequences of at least three cardiac cycles in which systolic blood pressure (SBP) and R-R interval changed in the same direction. Venous blood was collected during 90 degrees HUT and analyzed for changes in plasma NE concentrations, as well as hematocrit and hemoglobin to determine changes in PV. RESULTS: Endurance training significantly (P < 0.01) increased VO(2peak) (mL x kg(-1) x min(-1)) in EX by 10 +/- 2%. The immediate, early steady-state, and prolonged HR and AP responses and spontaneous baroreflex slopes during 90 degrees HUT were not significantly different (P > 0.05) between EX and CON groups before or after 4, 8, or 12 wk of endurance training. No significant differences (P > 0.05) were observed between EX and CON groups for peak changes in PV during orthostasis before (-15.0 +/- 1.4% vs -11.9 +/- 1.3%) or after 4 (-12.2 +/- 1.0% vs -12.7 +/- 1.4%), 8 (-13.7 +/- 1.2% vs -12.4 +/- 0.7%), and 12 wk (-10.8 +/- 1.6% vs -10.6 +/- 0.6%) of endurance training, suggesting a similar stimulus presented by 90 degrees HUT in both groups. Peak changes in NE concentrations during HUT were similar (P > 0.05) between EX and CON groups before (119 +/- 23 pg x mL(-1) vs 191 +/- 36 pg x mL(-1)) and after 4 (139 +/- 29 pg x mL(-1) vs 146 +/- 25 pg x mL(-1)), 8 (114 +/- 32 pg x mL(-1) vs 182 +/- 41 pg x mL(-1)), and 12 wk (143 +/- 35 pg x mL(-1) vs 206 +/- 42 pg.mL-1) of endurance training. CONCLUSIONS: These data indicate that in healthy elderly men, improvements in VO(2peak) can occur without compromising the regulation of HR, AP, NE, and PV during orthostatic stress. PMID- 11474328 TI - Exercise training and autonomic nervous system activity in obese individuals. AB - PURPOSE: This study was designed to investigate the effects of 12 wk of exercise training on autonomic nervous system (ANS) in 18 obese middle-aged men (N = 9) and women (N = 9) (age: 41.6 +/- 1.2 yr; BMI: 27.3 +/- 0.4 kg x m(-2); %fat: 29.6 +/- 1.3%, mean +/- SE). METHODS: Each subject participated in an aerobic exercise training at anaerobic threshold (AT), consisting of 30 min/session, 3 times/wk, for 12 consecutive weeks. The ANS activities were assessed by means of power spectral analysis of heart rate variability (HRV) at resting condition before, at 5 wk, and after the exercise program. RESULTS: The exercise training resulted in a significant decrease in body mass, BMI, and % fat (P < 0.01) but not in lean body mass (P > 0.05) together with a significant increase in the AT VO2 (P < 0.01). Our power spectral data indicated that there were significant increases in the low-frequency component associated with the sympathovagal activity (0.03- 0.15 Hz, 348.5 +/- 66.8 vs 694.7 +/- 91.5 ms(2), P < 0.01), the high-frequency vagal component (0.15--0.4 Hz, 146.3 +/- 30.4 vs 347.7 +/- 96.5 ms(2), P < 0.05), and the overall autonomic activity as evaluated by total power (0.03--0.4 Hz, 494.8 +/- 88.5 vs 1042.4 +/- 180.9 ms(2), P < 0.01) of HRV after the training. CONCLUSIONS: Twelve weeks of exercise training has significantly improved both the sympathetic and parasympathetic nervous activities of the obese individuals with markedly reduced ANS activity, suggesting a possible reversal effect of human ANS functions. These favorable changes may also have an influence on the thermoregulatory control over the obesity. PMID- 11474329 TI - Dietary restraint, exercise, and bone density in young women: are they related? AB - PURPOSE: Women with high scores for dietary restraint have been found to have higher 24-h urinary cortisol excretion and a higher prevalence of subclinical ovulatory disturbances, both of which may be risk factors for bone loss. The purpose of this study was to explore relationships between dietary restraint and bone health in regularly menstruating young women. METHODS: 62 women (age: 21.7 +/- 2.5 yr) had body composition and total body and lumbar spine bone mineral density (BMD) and content (BMC) assessed using dual-energy x-ray absorptiometry. Dietary restraint was assessed using the restraint subscale from the Three-Factor Eating Questionnaire: 29 women had low restraint (LR; restraint score 0--5), 33 had high restraint (HR; restraint score 13--21). Exercise (h x wk(-1)) was assessed by questionnaire on two occasions. RESULTS: LR and HR women were similar in age and body composition (fat mass = 15.0 +/- 4.7 kg, lean mass = 40.9 +/- 4.9 kg), but HR women exercised more (3.4 +/- 1.7 vs 2.2 +/- 1.8 h x wk(-1), P < 0.05). Exercise was correlated with BMD and BMC, and when it was included as a covariate, total body BMC was significantly lower in HR than LR women. In multiple regression analysis, weekly hours of exercise and restraint score were significant predictors of total body BMD and BMC. CONCLUSION: The observations of this cross-sectional study suggest that high levels of cognitive dietary restraint, or associated factors such as higher cortisol, may attenuate the positive effects of exercise on bone in young women. PMID- 11474330 TI - Muscular characteristics of detraining in humans. AB - Skeletal muscle is characterized by its ability to dynamically adapt to variable levels of functional demands. During periods of insufficient training stimulus, muscular detraining occurs. This may be characterized by a decreased capillary density, which could take place within 2--3 wk of inactivity. Arterial-venous oxygen difference declines if training stoppage continues beyond 3--8 wk. Rapid and progressive reductions in oxidative enzyme activities bring about a reduced mitochondrial ATP production. The above changes are related to the reduction in VO(2max) observed during long-term training cessation. These muscular characteristics remain above sedentary values in the detrained athlete but usually return to baseline values in recently trained individuals. Glycolytic enzyme activities show nonsystematic changes during periods of training cessation. Fiber distribution remains unchanged during the initial weeks of inactivity, but oxidative fibers may decrease in endurance athletes and increase in strength-trained athletes within 8 wk of training stoppage. Muscle fiber cross sectional area declines rapidly in strength and sprint athletes, and in recently endurance-trained subjects, whereas it may increase slightly in endurance athletes. Force production declines slowly and in relation to decreased EMG activity. Strength performance in general is readily maintained for up to 4 wk of inactivity, but highly trained athletes' eccentric force and sport-specific power, and recently acquired isokinetic strength, may decline significantly. PMID- 11474331 TI - Dietary creatine supplementation and muscular adaptation to resistive overload. AB - PURPOSE: This study examined the influence of dietary creatine (CR) supplementation upon mechanical and hypertrophic responses to a well-defined conditioning stimulus provided by electromyostimulation (EMS). METHODS: Eighteen resistance-trained subjects were assigned CR or a placebo (PL) in a randomized, double-blind fashion. After CR loading (20 g x d(-1) for 7 d), CR supplementation (5 g x d(-1)) or PL was continued for 8 wk. During supplementation, EMS (3--5 sets of 10 coupled eccentric and concentric actions) was applied to the left m. quadriceps femoris (QF) twice weekly while subjects continued voluntary resistance training of both lower limbs unsupervised. Cross-sectional area (CSA) of each QF was assessed with magnetic resonance imaging (MRI). Torque during EMS was analyzed to assess muscle loading and fatigue resistance. RESULTS: Maximal torque and the torque time integral increased markedly over training (P < or = 0.0001). These responses reflected activation of more muscle as EMS current was increased (about 16%), greater recovery between sets (P < or = 0.0423), and less fatigue during sets over training (P = 0.0002). CR did not influence these responses (P = 0.8093). In accord with these results, the increase in CSA for the stimulated QF (11%) was comparable for CR and PL (P = 0.2190). CSA in the nonstimulated QF increased 5% in CR (P = 0.0091) but did not change in PL. CONCLUSION: We conclude that CR supplementation did not augment the mechanical or hypertrophic response to a precisely measured conditioning stimulus that attenuated but did not ameliorate fatigue. We suggest that enhanced fatigue resistance may not explain the apparent ergogenic effect of CR during voluntary training. PMID- 11474332 TI - Use of air displacement plethysmography for estimating body fat in a four component model. AB - PURPOSE: To compare measurements of body density (D(b)) obtained from air displacement plethysmography (AP) and hydrostatic weighing (HW) and to determine the accuracy of substituting D(b) via AP (D(b)-AP) for D(b) via HW (D(b)-HW) in estimating body fatness (%Fat(4C)) and the composition and density of the fat free mass (Dffm) from a four-component model (fat, mineral, water, and protein). METHODS: D(b) was measured in 50 young adults using AP and HW. Total body water via deuterium dilution, bone mineral content via dual-energy x-ray absorptiometry, and D(b) were used to estimate %Fat(4C). RESULTS: D(b)-AP and D(b)-HW were highly correlated (r = 0.89, SEE = 0.008 g x mL(-1)), but D(b)-AP (1.065 +/- 0.003 g x mL(-1)) was significantly higher (P < 0.05) than D(b)-HW (1.058 +/- 0.003 g x mL(-1)), resulting in a mean difference of 2.8%fat. Differences between %Fat(4C-AP) (17.8 +/- 1.2%) and %Fat(4C-HW) (19.3 +/- 1.2%) were significant (P < 0.05), but the SD of the differences (2.3%) was low. When D(b)-AP was used in a four-component model in place of D(b)-HW, the calculated Dffm was significantly higher (1.109 +/- 0.002 vs 1.105 +/- 0.002 g x mL(-1)) based on a higher (P < 0.05) protein fraction (22.0 +/- 0.4% vs 20.6 +/- 0.4%) and lower (P < 0.05) water (71.1 +/- 0.4% vs 72.4 +/- 0.4%) and mineral fractions (7.0 +/- 0.1% vs 7.1 +/- 0.1%). CONCLUSIONS: AP yields a higher D(b) than HW and may not be a valid method for measuring D(b) or estimating %fat using densitometry. However, due to relatively small bias and low individual error, D(b)-AP is an acceptable substitute for D(b)-HW when estimating %fat with a four component model in young adults. PMID- 11474333 TI - Effect of impact load and active load on bone metabolism and body composition of adolescent athletes. AB - PURPOSE: It is unclear whether adolescents involved in nonweight-bearing activities experience a delay in bone growth acquisition and sexual maturation. The purpose of this study was to compare bone mineral density (BMD), body composition, hormonal profile, and bone biochemical markers of adolescent athletes active in sports involved in impact load sports with those participating in active load sports. METHODS: Forty-five male Caucasian athletes aged 12--18 yr were divided into two groups according to type of skeleton loading, impact (N = 18), or active (N = 27). Twenty-four male Caucasian adolescents (12--18 yr) served as controls and only performed the activities included in their physical education classes. All subjects were assessed for bone mass, body composition, and bone age by dual x-ray absorptiometry (DXA). Serum calcium (Ca), phosphorus (P), bone-specific alkaline phosphatase (BAP), total testosterone, FSH, LH, urinary calcium to creatinine ratio (Ca/Cr), and urinary deoxypyridinoline crosslinks to creatinine ratio (DPD/Cr) were measured. RESULTS: The impact load group presented the highest BMD among the three groups for all studied sites. Lean mass and absolute weight were correlated with all of the bone mass measurements. BAP levels were significantly higher and testosterone levels significantly lower in the active load group compared with the impact group. CONCLUSION: High-impact load exercises have a beneficial effect on bone mass in male adolescents. There is also a positive correlation of weight and body composition with BMD. However, further longitudinal studies are necessary to determine whether there is a delay in bone growth acquisition among adolescents involved in a nonweight-bearing exercise regimen and its association with sex hormones. PMID- 11474334 TI - Relationship between subcutaneous fatness and leptin in male athletes. AB - PURPOSE: Circulating leptin is low in trained subjects and closely related to body fat content. However, data are scarce as to whether differences exist in the relationship between different estimates of adiposity, metabolic parameters, and leptin in endurance- and resistance-trained male athletes. We investigated this relationship with special emphasis on subcutaneous fatness and its distribution. METHODS: 20 endurance (ET) and 17 resistance (RT) athletes recruited from different kind of sports were studied. Fat-free mass (FFM) was estimated by means of impedance and fat mass (FM) was calculated. Subcutaneous fat (SAT) and its distribution was measured by means of the optical device Lipometer at 15 body sites (SAT-layers; from 1-neck to 15-calf) on the right side of the body. Fifteen SAT-layers were summed to calculate SAT. Blood samples were obtained for determination for leptin, insulin, and glucose. Insulin resistance was calculated through the fasting insulin resistance index (FIRI; [insulin x glucose/25]). RESULTS: RT-athletes had a greater body mass and body fat content than ET athletes, but no differences were found for leptin and metabolic parameters. In all athletes, estimates of adiposity were correlated to leptin. However, in ET athletes FM (P < 0.05), FFM (P < 0.05), and SAT (P < 0.001) but not metabolic parameters were correlated to leptin. In RT-athletes, SAT (P < 0.0001), metabolic parameters (all P < 0.05), but not FM and FFM were in significant relationship with leptin. Stepwise regression revealed SAT as the main determinant for the variation in leptin in all athletes (adj. R(2 )= 0.52, P < 0.0001). CONCLUSION: The results suggest that estimates of adiposity and metabolic parameters are associated with leptin in a sport-specific manner. Whereas leptin might be regulated by overall subcutaneous fatness in athletes, our study does not imply a main influence of fat patterning on leptin in this group of trained subjects. PMID- 11474335 TI - Biomechanical factors affecting running economy. AB - PURPOSE: The present study was designed to investigate kinematics, kinetics, and muscle activity for explaining running economy at different running speeds. METHODS: A total of 17 young endurance runners ran at 12-13 different running speeds. Respiratory gases were collected. Kinematic records were obtained by a high-speed video camera, and 3-D ground reaction forces (GRF) were measured simultaneously with telemetric EMG recordings of the selected leg muscles. In the analysis, joint moments and power were calculated by inverse dynamic methods. RESULTS: The oxygen consumption and energy expenditure increased quite linearly with increasing running speed. However, already at the slowest speed, interindividual differences in running economy were noticed, and they increased with increasing running speed. Simultaneously, the instantaneous joint moment angular velocity curves of the ankle and knee joints shifted to the right and upward, thus increasing joint power in the push-off phase of contact. Most definitive was the increase in EMG-activity of the BF muscle and its correlation with energy expenditure (r = 0.48, P < 0.05). This two-joint muscle seems to be very active during the maximal running: its amplitude increased (P < 0.05) both in the swinging and contact phases with increasing running speed. CONCLUSIONS: The increased EMG of working muscles and the associated increase in power output may partly explain the increased energy expenditure with increasing running speed. Lower performances in running economy by some of the athletes may also be explained by poor running technique, such as unusually high braking and mediolateral forces, which may be caused by limited action of the hamstring muscles. However, no exclusive biomechanical parameters could be identified to explain the running economy. PMID- 11474336 TI - Ankle biomechanics during four landing techniques. AB - PURPOSE: An understanding of landing techniques is important for the prevention of injuries in a number of athletic events. There is a risk of injury to the ankle during landings, and the kinematics and forces involved in different landing strategies may be related to the occurrence of trauma. METHODS: In the current study, four drop conditions from a 30.48-cm (12-inch) height were tested. The conditions were a) BN: Bent knee (self-selected), Natural (self-selected) plantar flexor contraction; b) SN: Stiff-knee, Natural plantar flexors; c) SP: Stiff-knee, Plantar flexors absorbing the impact; and d) SH: Stiff-knee, absorbing most of the impact in the Heels. Peak vertical forces and accelerations were measured, and Achilles tendon forces and stiffnesses were calculated. RESULTS: Peak vertical forces and peak tibial accelerations were highest for the SH condition (2418 N and 20.7 G), whereas peak Achilles tendon force was highest for SP drops. The overall average AT stiffness was 166,345 N x m(-1). CONCLUSIONS: The results from the study were used in an extensive cadaver study to investigate ankle injuries. The data from the current study indicate that athletes may not use their full energy absorbing potential in landings during sporting activities. PMID- 11474337 TI - Biomechanical analysis of the deadlift during the 1999 Special Olympics World Games. AB - PURPOSE: Improper lifting techniques may increase injury risks and decrease performance. The aim of this study was to compare and contrast biomechanical parameters between sumo and conventional style deadlifts and between high- and low-skilled lifters who participated in the powerlifting event during the 1999 Special Olympics World Games. METHODS: Two synchronized video cameras collected 60 Hz of data from 40 subjects. Parameters were quantified at barbell liftoff (LO), when the barbell passed the knees (KP), and at lift completion. RESULTS: Compared with the conventional group, the sumo group had a 100% greater stance width, 20% smaller hand width, 10% less vertical bar distance, a more vertical trunk at LO, a more horizontal thigh at LO and KP, a less vertical shank at KP, and greater forefoot abduction. The sumo group generated ankle dorsiflexor, knee extensor, and hip extensor moments, whereas the conventional group produced ankle plantar flexor, knee flexor and extensor, and hip extensor moments. Compared with low-skilled lifters, high-skilled lifters had a 40% greater barbell load, 15% greater stance width (sumo group only), greater knee flexion at LO (conventional group only), greater knee extension at KP, a less vertical shank position at LO (sumo group only), 15% less vertical bar distance, less first peak bar velocity between LO and KP (conventional group only), smaller plantar flexor and hip extensor moment arms at LO and KP, and greater knee extensor moment arms at LO. CONCLUSIONS: The sumo deadlift may be more effective in working ankle dorsiflexors and knee extensors, whereas the conventional deadlift may be more effective in working ankle plantar flexors and knee flexors. High-skilled lifters exhibited better lifting mechanics than low-skilled lifters by keeping the bar closer to the body, which may both enhance performance and minimize injury risk. PMID- 11474338 TI - Neural adaptations to fatigue: implications for muscle strength and training. AB - PURPOSE: This paper investigates the neural mechanisms responsible for the increase in strength that occurs during serial isometric contractions. METHODS: A three-session design was used. Thirteen subjects (N = 13) were asked to perform five maximal isometric elbow extension strength trials to serve as baseline. After a 5-min rest, the subjects were administered a 30-trial fatigue protocol. This process was repeated two more times at 2-wk intervals. Elbow extension torque and surface electromyography (EMG) of the triceps and biceps brachii were monitored concurrently. The criterion measures were elbow extension torque, root mean-square EMG amplitude, and mean power frequency (MPF). RESULTS: Intraclass reliability ranged from good to excellent. Within each experimental session, the fatigue protocol resulted in a decrease in maximal isometric elbow extension torque as well as biceps and triceps EMG amplitude and MPF (P < 0.05). However, the mean of the 30 trials and the magnitude of the linear decrease in elbow extension torque increased across the three sessions (P < 0.05). Biceps and triceps EMG amplitude increased and MPF decreased as the number of sessions increased (P < 0.05). CONCLUSIONS: These findings suggest that the fatigue protocol served as a training stimulus to down regulate motor-unit firing frequency. PMID- 11474339 TI - Preferred pedalling cadence in professional cycling. AB - PURPOSE: The aim of this investigation was to evaluate the preferred cycling cadence of professional riders during competition. METHODS: We measured the cadence of seven professional cyclists (28 +/- 1 yr) during 3-wk road races (Giro d'Italia, Tour de France, and Vuelta a Espana) involving three main competition requirements: uphill cycling (high mountain passes of approximately 15 km, or HM); individual time trials of approximately 50 km on level ground (TT); and flat, long ( approximately 190 km) group stages (F). Heart rate (HR) data were also recorded as an indicator of exercise intensity during HM, TT, and F. RESULTS: Mean cadence was significantly lower (P < 0.01) during HM (71.0 +/- 1.4 rpm) than either F and TT (89.3 +/- 1.0 and 92.4 +/- 1.3 rpm, respectively). HR was similar during HM and TT (157 +/- 4 and 158 +/- 3 bpm) and in both cases higher (P < 0.01) than during F (124 +/- 2 bpm). CONCLUSION: During both F and TT, professional riders spontaneously adopt higher cadences (around 90 rpm) than those previously reported in the majority of laboratory studies as being the most economical. In contrast, during HM they seem to adopt a more economical pedalling rate (approximately 70 rpm), possibly as a result of the specific demands of this competition phase. PMID- 11474340 TI - Physiological and performance responses to tournament wrestling. AB - PURPOSE: The purpose of this study was to investigate the physiological and performance responses to a simulated freestyle wrestling tournament after typical weight loss techniques used by amateur wrestlers. METHODS: Twelve Division I collegiate wrestlers (mean +/- SD;19.33 +/- 1.16 yr) lost 6% of total body weight during the week before a simulated, 2-d freestyle wrestling tournament. A battery of tests was performed at baseline and before and immediately after each individual match of the tournament. The test battery included assessment for body composition, reaction/movement time, lower and upper body power and isokinetic strength, and a venous blood sample. RESULTS: Lower body power and upper body isometric strength were significantly reduced as the tournament progressed (P < or = 0.05). Significant elevations in testosterone, cortisol, and lactate were observed after each match (P < or = 0.05). However, there was a significant reduction (P < or = 0.05) in resting testosterone values in the later matches. Norepinephrine increased significantly (P < or = 0.05) after each match, whereas epinephrine increased significantly (P < or = 0.05) after each match except the last match of each day. Plasma osmolality was consistently higher than normal values at all times including baseline, with significant increases observed after each match (P < or = 0.05). CONCLUSIONS: Tournament wrestling augments the physiological and performance decrements of weight loss and its impact is progressive over 2 d of competition. The combined effects of these stresses may ultimately be reflected in a wrestler's ability to maintain physical performance throughout a tournament. PMID- 11474341 TI - Physiological determinants of cross-country ski racing performance. AB - PURPOSE: Previous laboratory testing has identified the importance of upper-body aerobic and anaerobic power to cross-country skiing performance. The purpose of this investigation was to extend laboratory research into a field setting to identify predictors of performance through ski-specific testing. METHODS: Thirteen male collegiate skiers performed three field-testing sessions on roller skis to establish lactate threshold (LT) and ski economy (ECON) and maximal oxygen uptake (SK VO(2max)) and a 1-km double-pole time trial (UBTT) to determine peak upper-body oxygen uptake (UB VO(2)). As a measure of skiing performance, the subjects performed a 10-km skating time trial (TT) and were ranked according to competitive season performance (RANK). RESULTS: Significant correlations (P < 0.05) were found between SK VO(2max), LT VO(2), UB VO(2), and RANK (r = -0.66 to 0.84) and TT time (r = -0.74 to -0.79), as well as ECON to RANK (r = 0.57) and TT time (r = 0.68). Time to complete the UBTT (UB time) exhibited the strongest correlation to both RANK (r = 0.95) and TT time (r = 0.92). Multiple regression analyses revealed that UB time was the best predictor of RANK and TT time, as demonstrated by the significant beta values (0.77, P < 0.001, and 0.79, P < 0.001, respectively). The importance of the UB component was further seen in that UB time was still the best predictor of performance when the subjects were divided into two distinct groups of greater and lesser competitive ability. CONCLUSIONS: These findings identify the importance of the upper body component to cross-country skiing performance, suggesting a need to focus on upper-body conditioning within a well-rounded endurance training program. Additionally, the UBTT exhibits potential as a simple field test to predict cross-country skiing performance over more sophisticated and costly laboratory and field testing. PMID- 11474342 TI - Ultraviolet exposure in the Ironman triathlon. AB - PURPOSE: Skin cancer is increasing worldwide and exposure to ultraviolet (UV) radiation is thought to be the most important environmental risk factor. People practicing outdoor sports are exposed to considerable amounts of UV radiation from the sun. METHODS: Three triathletes participated in the Ironman Triathlon World Championships 1999 in Hawaii (3.9-km swim, 180.2-km bike, 42.4-km run). They attached Bacillus subtilis spore film dosimeters (VioSpor) on the back between their shoulders. The dosimeter system measured cumulative biologically weighted erythemal UV exposure. UV exposure is given in minimal erythema doses (1 MED corresponds to 250 J x m(-2) at 298 nm). RESULTS: The mean personal UV exposure was 8.3 MED (6.9--9.7 MED) after 8:43 to 9:44 h of competition corresponding to 0.8 to 1.3 MED x h(-1) (bike and run). The athletes were sunburned despite the use of water-resistant sunscreen (SPF 25+) on sun exposed skin. CONCLUSION: The International Radiation Protection Agency has issued guidelines for professional UV exposure. Ironman triathletes considerably exceeded these limits of exposure similar to other outdoor sports. Professional and amateur athletes should be aware of hazards caused by UV radiation. Adequate protection by water-resistant sunscreens and clothing as well as training and competition schedules with low sun exposure seem to be a reasonable recommendation. PMID- 11474343 TI - Aerobic and anaerobic power characteristics of off-road cyclists. AB - PURPOSE: The purpose of this study was to describe the relationship between anaerobic power at different pedaling frequencies (including the optimal cadence) and aerobic power in off-road cyclists (CYC; N = 25) and sports students, who did not perform specific cycle exercise more than two times per week (CON; N = 60). METHODS: To describe the aerobic power, we measured the maximal power output (W(max)) and the power output at the fixed lactate threshold at 4 mmol x L(-1) (W(L4)) obtained during a maximal aerobic power cycling test. To describe anaerobic power output, we measured the average power output (IsoW(mean)) over a range from 50 to 140 rpm by using a 10-s sprint on an isokinetic cycle ergometer. RESULTS: For the 10-s anaerobic test, CON and CYC showed a peak power output (IsoW(peak)) of 13.3 +/- 1.4 and 14.9 +/- 1.1 W x kg(-1), respectively. IsoW(peak) corresponded to an optimal cadence of 100 +/- 9.3 rpm for CON and 100 +/- 8.7 rpm for CYC. There was a significant difference (P < 0.001) in the W(max):IsoW(peak) (W(aerobic):W(anaerobic)) ratio between CON (32 +/- 4.5%) and CYC (38 +/- 3.9%). Significant differences among group means were identified using an ANOVA test and a post hoc analysis. The off-road cyclists showed a significantly higher IsoW(mean) at all pedaling frequencies and at the optimal cadence (P < 0.01). There was a modest relationship between W(max) and IsoW(peak) in both groups (CON r = 0.53; CYC r = 0.64; P < 0.01). CONCLUSION: Anaerobic power values are important components associated with cycle performance in both noncyclists and off-road cyclists. However, the results of the present study demonstrated the usefulness of the power index in the physiological evaluation of off-road cyclists, as it gives information on the proportion of aerobic to anaerobic energy contribution. PMID- 11474344 TI - Heart rate variability in athletes: relationship with central and peripheral performance. AB - PURPOSE: To investigate relationships between heart rate variability (HRV) and peripheral and central performance measures, 17 cross-country (X-C) skiers and seven canoeists were studied before and after a training period of 7 months. METHODS: For the skiers and canoeists respectively, leg and arm peak torque (Tq), time to peak torque (TiTq), and total work (Wrk) were measured in an isokinetic dynamometer. Maximal oxygen uptakes (VO(2max)) were obtained from treadmill tests. Power spectral analysis of HRV was performed on electrocardiographic recordings in the resting supine position and after a tilt to yield power in the low-frequency (0.04--0.15 Hz) and high-frequency (0.15--0.45 Hz) components of HRV. RESULTS: The change in normalized LF-variability in standing (Delta LFnT) correlated (P < 0.01) with the changes in TiTq (r = 0.63), max lactate (r = 0.63), and VO(2max) (r = -0.53). The change in absolute LFT was inversely correlated with the change in Tq. Subjects who improved VO(2max) were characterized by consistently higher high-frequency and total HRV than subjects with deteriorated aerobic capacity (P < 0.05). CONCLUSION: The results suggest that improved measures of both peripheral and central (aerobic) work capacities were associated with a reduction of low-frequency HRV in the tilted position. High-frequency and total HRV did not change in proportion with changes in muscle performance or aerobic capacity, but the ability to further improve VO(2max) with training in these already fit subjects seemed to depend on their average levels of these HRV measures, interpreted to reflect parasympathetic activity. PMID- 11474345 TI - Effect of caffeine and ephedrine ingestion on anaerobic exercise performance. AB - PURPOSE: Ingestion of a combination of caffeine (C) and ephedrine (E) prolongs time to exhaustion during high-intensity aerobic exercise. CNS stimulation by C and E was proposed as part of the mechanism for the improvement. It was thought that this arousal might also be of benefit during anaerobic exercise. The purpose of this study was to investigate the effect of C, E, and C+E ingestion on performance of anaerobic exercise. METHODS: Two groups were used to evaluate the effect of C and E on anaerobic performance. Group 1 (WIN) consisted of 16 healthy untrained male subjects who performed a 30-s Wingate test. Group 2 (MAOD) consisted of 8 healthy untrained male subjects who performed a supramaximal (125%VO(2peak)) cycle exercise trial to exhaustion to determine maximum accumulated oxygen deficit. The trials commenced 1.5 h after ingesting either C (5 mg x kg(-1)), E (1 mg x kg(-1)), a combination of C+E, or a placebo (P). All trials were randomized and double blind. Blood samples were assayed for lactate and glucose post drug ingestion just before exercise, and again 3, 5, and 10 min post exercise. Catecholamines were measured in the preexercise and 10-min postexercise blood samples. RESULTS: Ephedrine increased power output during the early phase of the Wingate test, whereas C increased time to exhaustion and O(2) deficit during the MAOD test. C, E, and C+E increased blood lactate, glucose, and catecholamine levels. CONCLUSION: The improvement in anaerobic exercise performance is likely a result of both stimulation of the CNS by E and skeletal muscle by C. PMID- 11474346 TI - Intermodal comparison of energy expenditure at exercise intensities corresponding to the perceptual preference range. AB - PURPOSE: This study compared the rate of energy expenditure among six popular exercise machines at intensities corresponding to ratings of perceived exertion (RPE) for fairly light (RPE-11), somewhat hard (RPE-13), and hard (RPE-15) in 9 healthy men and 10 healthy women. METHODS: A maximal exercise test on each exercise machine was used to anchor the Borg 15-point category scale. Subjects performed three submaximal exercise tests at selected RPEs on a treadmill, stair stepper, cycle ergometer, rowing ergometer, cross-country ski simulator, and rider. The submaximal tests on each exercise device were performed in random order and were 6 min in duration with 15-min rest between trials. Oxygen uptake, heart rate, and blood lactate concentration were measured during the final 2 min of each exercise intensity. RESULTS: Energy expenditure at each RPE was highest on the treadmill and ski simulator in men, and on the treadmill, ski simulator, and rowing ergometer in women. Energy expenditure in men and women at all RPEs was lowest on the rider and cycle ergometer. Energy expenditure at a given RPE was greater in men than women on all exercise machines, but men and women used a similar percentage of their machine specific peak oxygen uptake at each RPE on all machines. Heart rate was generally similar among the machines and between both men and women at each RPE. CONCLUSIONS: Our results indicated that there are large differences in energy expenditure between exercise machines and between men and women at intensities perceived to be fairly light, somewhat hard, and hard. Consequently, subjects can expend more calories at the same RPE during treadmill and ski simulator exercise, for example, than during exercise with other devices. This may have important implications for the health benefits of different exercises and in promoting long term exercise adherence. PMID- 11474347 TI - Differences between whole blood and plasma lactate concentrations have to be considered when comparing various studies. PMID- 11474348 TI - A multicenter, prospective, randomized trial evaluating a new hemostatic agent for spinal surgery. AB - STUDY DESIGN: A prospective, randomized trial comparing Proceed, a gelatin-based hemostatic sealant (treatment), with Gelfoam-thrombin (control) in stopping intraoperative bleeding during spinal surgery. OBJECTIVES: To determine the effectiveness and safety of Proceed. SUMMARY OF BACKGROUND DATA: Proceed has been tested in animal models to determine its safety and effectiveness as a hemostatic agent. The current study was conducted under a Food and Drug Administration approved Investigational Device Exemption to evaluate the effectiveness and safety of Proceed in humans. METHODS: For this study, 127 patients undergoing spinal surgery were randomized into either the treatment or control group after standard surgical means to control bleeding had failed. The bleeding site was evaluated at 1, 2, 3, 6, and 10 minutes after the hemostatic agent was applied. The application was considered successful if the bleeding stopped within 10 minutes. Follow-up evaluation was performed at 12 to 36 hours, then at 6 to 8 weeks after surgery. RESULTS: Proceed stopped bleeding in 98% of the patients (first bleeding site only) within 10 minutes, as compared with 90% of the control patients (P = 0.001). At 3 minutes, successful hemostasis had been achieved in 97% of the Proceed group, as compared with 71% of the control group (P = 0.0001). There was no difference in the adverse event profile between the two groups. CONCLUSIONS: A significantly larger number of bleeding sites had achieved hemostasis with Proceed than with Gelfoam-thrombin at 1, 2, and 3 minutes after application. Proceed was as safe as Gelfoam-thrombin when used for hemostasis during spinal surgery procedures. PMID- 11474349 TI - Effects of anulus fibrosus and experimentally degenerated nucleus pulposus on nerve root conduction velocity: relevance of previous experimental investigations using normal nucleus pulposus. AB - STUDY DESIGN: Nerve conduction velocity was measured in the pig cauda equina after local application of anulus fibrosus or in vitro/postmortem degenerated nucleus pulposus from the same pig. OBJECTIVES: To analyze the effects of anulus fibrosus and degenerated nucleus pulposus on nerve conduction velocity. SUMMARY OF BACKGROUND DATA: Previous studies on nucleus pulposus-induced effects on nerve roots have used normal, nondegenerated nucleus pulposus. Because both anulus fibrosus and degenerated nucleus pulposus are commonly seen in the clinical situation of disc herniation, the value of the previous work could be questioned. METHODS: Anulus fibrosus and nucleus pulposus were harvested using a retroperitoneal approach. The nucleus pulposus was degenerated artificially either by addition of sodium lactate with HCl added to form a pH of either 6.0 or 3.5 (in vitro degeneration), or by storing the nucleus pulposus at 4 C until a pH of 6.0 (postmortem degeneration) was reached. After epidural application, the nerve conduction velocity was determined at 7 days (anulus fibrosus) or 3 days (degenerated nucleus pulposus). RESULTS: Application of anulus fibrosus did not induce any reduction of nerve conduction velocity. In vitro and postmortem degenerated nucleus pulposus induced a reduction of nerve conduction velocity similar to that of normal nucleus pulposus. CONCLUSIONS: Although only nerve function and not pain was assessed, it seems likely that previous experiments using normal nucleus pulposus may be relevant for evaluating the pathophysiologic mechanisms behind the nucleus pulposus-induced nerve root injury, also in a clinical perspective. PMID- 11474350 TI - Osteogenic protein-1 overcomes the inhibitory effect of nicotine on posterolateral lumbar fusion. AB - STUDY DESIGN: An established rabbit posterolateral lumbar fusion model was used to evaluate the ability of osteogenic protein-1 to overcome the inhibitory effect of nicotine. OBJECTIVE: To determine whether osteogenic protein-1 should be considered as a bone graft alternative for the patient who smokes. SUMMARY OF BACKGROUND DATA: Smoking interferes with the success of posterolateral lumbar fusion. This inhibitory effect has been attributed to nicotine and confirmed in a New Zealand white rabbit model. Osteoinductive protein-1 has been shown to induce posterolateral spine fusion reliably in the rabbit model. The effectiveness with which osteogenic protein-1 induces fusion in the presence of nicotine has not been studied previously. METHODS: Single-level posterolateral intertransverse process fusions were performed at L5-L6 in 18 New Zealand white rabbits. Either autograft or osteogenic protein-1 was used as grafting material. Nicotine was administered via subcutaneous mini-osmotic pumps. The animals were killed 5 weeks after surgery, and the resulting fusion masses were studied. RESULTS: Three rabbits (17%) were excluded because of complications. By manual palpation, two of the eight nicotine-exposed autograft rabbits (25%) and all of the nicotine exposed osteogenic protein-1 rabbits (100%) were found to be fused. These results correlated well with those obtained from biomechanical testing. Histologically, the fusion zones of the nicotine-exposed autograft rabbits were distinctly less mature than the fusion masses of the nicotine-exposed osteogenic protein-1 rabbits. CONCLUSION: Osteoinductive protein-1 was able to overcome the inhibitory effects of nicotine in a rabbit posterolateral spine fusion model, and to induce bony fusion reliably at 5 weeks. PMID- 11474352 TI - Biomechanical comparison of posterior cervical fixation. AB - STUDY DESIGN: Biomechanical stability using four different posterior cervical fixation techniques was evaluated in human cadaveric spine. OBJECTIVES: To introduce an alternative interspinous fixation technique using wavy-shaped rods, and to compare its in vitro biomechanical stability with that of other posterior cervical fixation techniques. SUMMARY OF BACKGROUND DATA: Fixation of the posterior cervical spine with interspinous wiring is well known as Rogers' or Bohlman's technique. Recently, several plate fixation techniques have been used for posterior cervical stabilization. Since 1983, the authors have developed the wavy-shaped rod system as an alternative to the interspinous fixation technique. This unique technique has been proven clinically useful in Japan. However, the authors are not aware of any prior biomechanical studies. METHODS: Seven fresh frozen cervical human spines were tested at the C5-C6 motion segment. Nondestructive static biomechanical testing was performed with flexion-extension, lateral bending, and axial rotation for the following stabilization techniques: intact spine, creation of a Stage 3 distractive-flexion injury followed by fixation with the wavy-shaped rods bounded by three multistrand cables, interspinous wiring with a multistrand cable, triple wiring technique using multistrand cables with a pair of unicortical grafts from the ilium, and lateral mass plate fixation with Magerl's screw technique. Testing was performed on a material testing machine (MTS 858 Bionix test system, MTS, Minneapolis, MN), and load displacement curves were obtained using four linear extensometers and one rotatory extensometer across the C5-C6 motion segment. RESULTS: In axial compression loading, the reconstructed specimens showed significant differences in range of motion measured at the anterior and posterior positions, and statistical analysis was performed using one-way analysis of variance. In a comparison of the four fixation techniques, the construct with the wavy-shaped rod indicated significantly less motion both anteriorly and posteriorly than with the other fixation techniques. Also in flexion-extension loading, all the techniques significantly limited the intervertebral motion below the level of the intact motion segment. Particularly, the construct with the wavy-shaped rod showed the smallest mobility, 49.9% anteriorly and 9.3% posteriorly, compared with that of the intact spine. In lateral bending, the lateral mass plate provided the greatest stability, which was superior to the intact segment, but the difference was not statistically significant. In axial rotation, all the reconstruction techniques limited the angular motion below the intact level (wavy rod, 68.0%; Rogers' wiring, 75.2%; Bohlman's triple wiring, 59.8%; lateral mass plate, 71.7%), but no significant differences were observed using one-way analysis of variance, as compared with the intact segment. CONCLUSIONS: All four reconstruction techniques restored the stability of the cervical motion segment to at least the level of the intact motion segment before destabilization. An alternative cervical posterior fixation technique, the Wavy Rod system, was considered the most effective technique in stabilizing a cervical motion segment, particularly in axial compression and flexion extension loading. PMID- 11474353 TI - Notch sensitivity of titanium alloy, commercially pure titanium, and stainless steel spinal implants. AB - STUDY DESIGN: This is a biomechanical study comparing the fatigue life of stainless steel (SS), titanium alloy (TI), and commercially pure titanium (CP) rods that were notched using techniques essential to spine surgery. OBJECTIVES: To show the difference in fatigue life of the three rods and the effect on fatigue life of marks caused by techniques unavoidable in spine surgery. SUMMARY OF BACKGROUND DATA: Past studies have shown a decrease in the fatigue life of titanium when there is a defect or notch in the surface. This has been demonstrated with spinal and fracture implants, prosthetic joints, and in the materials science literature. Previous authors have recommended avoidance of techniques that mark the surface of titanium implants. METHODS: Four Texas Scottish Rite Hospital rods of each metal were prepared in four different ways: 1) no notch, 2) a French bender notch, 3) a top-loading connector--left on, 4) a top-loading connector--removed. The rods were fatigued in four point bending up to 1 million cycles. Before fatigue testing, two samples of each preparation were examined with electron microscopy. RESULTS: No SS rods failed. No TI rods without a notch or connector removed failed. One TI rod with the connector left in place failed (106,635 cycles). Three of the TI rods in the French-bender group failed (average, 87,663 cycles). One CP rod without a notch failed (358,957 cycles). No CP rods with the connector left in place failed. All CP rods with the connector removed failed (average, 230,094 cycles). All CP rods in the French bender group failed (average 120,817 cycles). The following differences were significant (P < 0.05): SS-TI and SS-CP in the French bender group; SS-CP and TI-CP in the connector removed group. Electron micrographs performed before fatigue testing showed cracks in the TI and CP rods that were marked with the French bender. CONCLUSION: Caution is recommended when bending TI or CP rods, or placing a connector and removing it when using CP rods in situations in which fatigue failure is a concern. PMID- 11474354 TI - Tumoral calcinosis of the spine: a study of 21 cases. AB - STUDY DESIGN: A retrospective study was conducted to investigate 21 patients found during spinal surgery to have paraspinal masses of dystrophic calcification (tumoral calcinosis). OBJECTIVES: To determine the magnetic resonance imaging characteristics of this disorder, and to document the associated spinal pathology. SUMMARY OF BACKGROUND DATA: Tumoral calcinosis usually is associated with hereditary disorders of calcium metabolism or renal dialysis. It also occurs in degenerated tissues in the absence of systemic disorders. Characteristically, calcific masses in the appendicular skeleton are visible on plain radiographs. Tumoral calcinosis has only rarely been reported in the spine. Documented patients have had an obvious calcific mass, and almost always the disorder has existed in other locations as well. Careful histologic study of specimens removed during spinal surgery suggests that tumoral calcinosis is common in the spine and usually is secondary to preexisting pathology. METHODS: This study involved 21 patients with lesions of tumoral calcinosis identified by histopathologic analysis of specimens removed during spinal surgery. The magnetic resonance images and the plain radiographs of the patients were reviewed and correlated with their clinical histories. RESULTS: In all the patients, the lesion of tumoral calcinosis was associated with a mass lesion seen on magnetic resonance imaging. Calcific masses were not apparent on plain films. In no case was the mass diagnosed before surgery as tumoral calcinosis. The magnetic resonance imaging changes were variously misinterpreted as neoplasms, infections, extruded disc material, or cysts. The observed features of tumoral calcinosis were those of an extradural mass showing a heterogeneous mixed-signal lesion that was identical on T1- and T2-weighted images. Characteristically, gadolinium did not enhance the lesions. CONCLUSIONS: Awareness of tumoral calcinosis of the spine may prevent unwarranted diagnoses of a more serious lesion in patients with characteristic magnetic resonance imaging changes. Also, this awareness may prevent pathologists from interpreting lesional tissue as nondiagnostic when other diagnoses are suspected clinically. This process may be a manifestation of degenerative spinal disease that has become so dominant that the underlying processes are obscured. PMID- 11474355 TI - Maximal versus feigned active cervical motion in healthy patients: the coefficient of variation as an indicator for sincerity of effort. AB - STUDY DESIGN: Maximal and submaximal (feigned) cervical motions in healthy patients were compared. OBJECTIVE: To test the efficiency of the coefficient of variation in differentiating maximal (sincere) from submaximal (feigned) cervical motion in healthy patients. SUMMARY OF BACKGROUND DATA: Although limitation of cervical motion is a recognized impairment, no well-founded approach for verifying the degree to which a patient is maximizing his or her performance is available currently. METHODS: A new ultrasound-based system for three-dimensional motion analysis of the head was used to test 25 healthy patients (22 women and 3 men) ages 26 to 48 years. During the first test, (Test 1), the patients were asked initially to move the head maximally at a self-determined velocity in all the primary directions: flexion, extension, right and left rotation, and right and left lateral flexion. They then were presented with a short vignette describing a fictitious accident and asked, using the same protocol, to perform the same types of cervical motions as if they had experienced an injury. No further instructions were provided. A retest (Test 2) in reverse order (feigned effort first) took place 1 to 16 weeks (mean, 3.3 weeks) later. RESULTS: The range of motion and average velocity were significantly smaller (P = 0.0001) in the feigned than in the maximal performance in all directions and on both tests. Feigned range of motion and velocity also were significantly reduced in Test 2 (P = 0.006), as compared with those in Test 1 (P = 0.0001). The range of motion coefficient of variation in the feigned performance (CVf) remained stable on the retest, but was significantly larger on both tests (P < 0.001) than the corresponding CV derived from maximal performance (CVm). Furthermore, a case-by case analysis showed that whereas the American Medical Association guidelines identified only 16% of the feigned cases, the use of CVf and CVm resulted in a corresponding rate of 87%. Tolerance intervals at 95% and 99% indicated total separation between the distributions of CVf and CVm, respectively. The average velocity-based CVf and CVm were not of a comparable differentiating power. CONCLUSIONS: This study indicates that in healthy patients, feigned performance may be differentiated from maximal (sincere) performance effectively and reproducibly using the coefficient of variation. PMID- 11474356 TI - Scoliosis in patients with diastrophic dysplasia: a new classification. AB - STUDY DESIGN: Scoliosis in patients with diastrophic dysplasia was analyzed. OBJECTIVES: To study the natural history of scoliosis and to classify the patients with different types of scoliosis. SUMMARY OF BACKGROUND DATA: Typical findings in diastrophic dysplasia are short-limbed short stature, multiple joint contractures, early degeneration of joints, and spinal deformities. The largest studies have reported scoliosis in 37% to 88% of the patients with this rare skeletal dysplasia. The natural history of the deformity is unknown. METHODS: Of the 130 unselected patients, 98 (75%) who were older than 16 years and/or had undergone surgery at the time of the last radiograph were included in the final analysis. These 98 patients included 37 males and 61 females. Their ages at the first radiograph ranged from newborn to 78 years (average, 21 years). The mean follow-up period was 20 years (range, 2-41 years) for 80 patients. Standard standing anteroposterior and lateral radiographs were taken. The degrees of scoliosis, kyphosis, and lordosis were measured according to the Cobb method. Classification of the scoliosis was based on the patient's age at onset of scoliosis, the rate of progression, the magnitude of the scoliosis at the end of growth, and the curve pattern. RESULTS: Of the 98 patients in this study, 86 (88%) had scoliosis. This difference was highly significant statistically (P < 0.001), as compared with the normal population. The frequency of scoliosis was 90% among females and 84% among males. Scoliosis can be divided further into three subtypes: early progressive (11 patients), idiopathic-like (41 patients), and mild nonprogressive (33 patients). One patient had a congenital scoliosis. CONCLUSIONS: Scoliosis is very common in patients with diastrophic dysplasia. The natural history of scoliosis varies from severe deformity with rapid progression to mild deformity without any progression. The authors suggest that the classification described in this report offers a tool for the predicting natural history of scoliosis in diastrophic dysplasia, and for adjusting the timing of surgery in individual patients. PMID- 11474357 TI - Early-phase enhanced inflammatory reaction after spinal instrumentation surgery. AB - STUDY DESIGN: The erythrocyte sedimentation rate, C-reactive protein, white blood cell count, and body temperature were measured prospectively in patients after two types of spinal surgery without complications and three cases of infection after spinal instrumentation surgery. OBJECTIVES: To investigate the effects of instrumentation on postoperative inflammatory reaction, and to describe early detection of postoperative wound infection. SUMMARY OF BACKGROUND DATA: In thoracic and abdominal surgery as well as hip arthroplasty, C-reactive protein has proved more valuable than erythrocyte sedimentation rate for early detection of postoperative infectious complications. It has not yet been established, however, how inflammatory parameters change after surgery when spinal instruments have been inserted into the body. METHODS: For this study, two groups of patients were examined: a control group that underwent spinal decompression surgery without instrumentation (n = 36) and another group that underwent spinal decompression and fusion surgery with spinal instrumentation (n = 37). The erythrocyte sedimentation rate, C-reactive protein, white blood cell count, and body temperature were recorded 1 day before surgery and on days 0 to 4, 7, 11, 14, 21, 28, and 42 after surgery. RESULTS: Inflammatory indexes (i.e., C-reactive protein, erythrocyte sedimentation rate, white blood cell count, and body temperature) were significantly higher for the surgery with instrumentation than for the spinal decompression surgery without instrumentation. Multiple regression analysis showed that C-reactive protein and erythrocyte sedimentation rate peaks significantly correlated with the use of instrumentation (C-reactive protein: P = 0.000257, erythrocyte sedimentation rate: P = 0.000132). In the patients with infection after spinal instrumentation surgery, C-reactive protein, white blood cell count, and body temperature started to increase again 4 to 11 days after surgery. The elevation of erythrocyte sedimentation rate levels was prolonged. CONCLUSIONS: Erythrocyte sedimentation rate and C-reactive protein display a significantly higher reaction after spinal surgery with instrumentation. Renewed elevation of C-reactive protein, white blood cell count, and body temperature after postoperative days 4 to 7 may be a critical sign of postoperative infection. PMID- 11474358 TI - Image-guided endoscopic spine surgery: Part I. A feasibility study. AB - STUDY DESIGN: A feasibility study was performed to determine the efficacy of computer assistance in endoscopic spine surgery. OBJECTIVES: To assess a new method for computer assistance based on image guidance during thoracoscopic or any endoscopic spine procedure. To evaluate the reproducibility, the sensitivity and the reliability of the technique first in vitro and second in clinical use. SUMMARY OF BACKGROUND DATA: The computer-based, image-guided surgery is now a routine tool used in open spine surgery. Exposure of the anatomy of the vertebra is needed for registration. This methodology is inapplicable in endoscopic approach. Fluoroscopic-based navigation combines the technology of image-guided surgery and C-arm fluoroscopy. The navigation is based on the fluoroscopic images acquired before surgery. This technology is applicable to endoscopic surgery but the navigation is based on fluoroscopic image. The computed tomography images are not exploited. There are no published data on a technique that allows image guided surgery based on computed tomography and magnetic resonance imaging. METHOD: A laboratory study was performed on a thoracic human spine. One vertebra was marked on the right lateral side of the body with five titanium marks. A percutaneous reference frame was specifically designed to be placed in the pedicle of the same marked vertebrae. The reference frame acted as a 3D localizer and a registration tool. The spine model was scanned including the reference frame. A standard Stealth station treatment guidance platform (Medtronic, Sofamor Danek, Memphis, TN) was used for simulation. The registration was obtained using the reference frame. Twenty navigation procedure trials were done and the error was recorded based on the distance between the anatomical point and the corresponding virtual one. RESULTS: Registration was always possible using the stealth station and a standard spine navigational software (spine 3, Medtronic Sofamor Danek, Memphis, TN). The mean error after registration given by the computer was 0.96 mm. The mean error recorded during the navigation simulation was 1.6 mm. CONCLUSIONS: This technique allows the possibility of computed tomography and magnetic resonance imaging-based, image-guided endoscopic surgery. It is probable that in the near future, as image fusion technology improves, the fluoronavigation based on fluoroscopic images would enable to navigate on multimodal images. Otherwise the technique described in this article is the only reproducible one that allows computed-tomography-based computer assistance during endoscopic procedures. PMID- 11474359 TI - Image-guided endoscopic spine surgery: Part II: clinical applications. AB - STUDY DESIGN: Endoscopic spinal procedures were performed under computed tomography-based, image-guided assistance. OBJECTIVE: To assess the clinical feasibility of applying a methodology that allows image-guided assistance in endoscopic spinal surgery. SUMMARY OF BACKGROUND DATA: Endoscopic spinal procedures have become a part of the minimal invasive approaches to the spine. The main disadvantage of these techniques is the long learning curve and the lack of peroperative monitoring. Fluoroscopy does have disadvantages, such as positioning during surgery and the risk for radiation exposure. Fluoroscopy-based navigation has many advantages, however it is still based on preselected fluoroscopic images. There is no method that allows computed-tomography-based navigation in endoscopic conditions. METHODS: Two patients have been operated on using endoscopic approaches assisted by computed-tomography-based navigational system. One had a thoracoscopic approach for median calcified disc herniation and another one had an endoscopic posterior approach for resection of a sacro-iliac osteophyte. For each patient, a frame of reference had been placed percutaneously and scanned. The computed tomography images were registered to the anatomy using the geometry of the frame as fiducials. Navigation through endoscopic approaches was possible in both cases. RESULTS: In both cases navigation was reliable and a helpful monitoring to achieve the surgical goals through endoscopic approaches. CONCLUSIONS: There are some factors that make endoscopic spine surgery a difficult start. Image-guided spine surgery is technically feasible and clinically applicable in endoscopic approaches. PMID- 11474360 TI - Patent foramen ovale and brain microembolization during scoliosis surgery in adolescents. AB - STUDY DESIGN: A case series is reported. OBJECTIVE: To improve understanding of the potential mechanisms associated with cerebral microemboli during scoliosis surgery in adolescents. SUMMARY OF BACKGROUND DATA: Paradoxical cerebral fat microembolization during scoliosis surgery has been associated with right-to-left shunting through an undetected patent foramen ovale. The prevalence of this cardiac defect in the adult population may be as high as 15% to 25%. Although the clinical relevance of this embolic phenomenon during scoliosis surgery has not been investigated, a few reports have documented its fatal consequences. It has been suspected in some patients with postoperative reduced visual function, particularly after complex instrumented fusions. METHODS: Bilateral transcranial Doppler (2 MHz) was used to monitor cerebral microemboli in the right and left middle cerebral arteries during spinal instrumentation and fusion in four patients. Additionally, transthoracic or transesophageal echocardiography incorporating pulse and color Doppler (3.5-5 MHz) and a contrast test was used in these patients to detect atrial shunts. RESULTS: Intraoperative transcranial Doppler monitoring detected high rates of cerebral microemboli associated with the presence of an atrial communication in two adolescents. In two additional patients with no detected brain microembolization, echocardiographic examination indicated the absence of an atrial shunt. CONCLUSION: The study findings suggest that unrecognized atrial communications in adolescents undergoing scoliosis surgery contribute to higher rates of Doppler-detected cerebral microemboli than those found in adolescents without this congenital defect. PMID- 11474361 TI - Posterior atlanto-occipital dislocation and concomitant discoligamentous C3-C4 instability with survival. AB - STUDY DESIGN: A case report is presented. OBJECTIVES: To describe the diagnostic procedure, treatment, and outcome of a 56-year-old man with posterior atlanto occipital dislocation and concomitant discoligamentous C3-C4 instability. CASE REPORT: A 56-year-old male seat-belted driver of a car was involved in an accident. After initial spontaneous breathing and weak movements of all his extremities, artificial respiration became necessary because of increasing respiratory insufficiency. Radiologic diagnostics, including computed tomography scans, showed a posterior atlanto-occipital dislocation and a Grade 2 craniocerebral trauma with occipital subarachnoidal bleeding. Further examination showed a serial rib fracture with concomitant hemopneumothorax. After stabilization of the patient, magnetic resonance imaging of the cervical spine showed a contusion of the upper cervical spinal cord. Additionally, rupture of the interspinal ligaments and the posterior longitudinal ligament could be seen, as well as a rupture of the intervertebral C3-C4 disc. Operative stabilization was performed by posterior fusion of C0-C4 using the CerviFix-System and autogenous bone grafts taken from the iliac crest. At 12 months after the operation, the patient had only slight weakness of the left arm and a 70% limitation in the range of motion of the cervical spine. CONCLUSIONS: With improvements in advanced trauma life support, the number of patients with atlanto occipital dislocation admitted to hospital alive is increasing, so this particular lesion must be kept in mind. When the patient survives the accident, the long-term prognosis is quite good, with a high rate of recovery after initial neurologic deficits. PMID- 11474362 TI - Spinal toxoplasmic arachnoiditis associated with osteoid formation: a rare presentation of toxoplasmosis. AB - STUDY DESIGN: An extremely rare presentation of an isolated spinal toxoplasmic arachnoiditis is described. OBJECTIVE: To draw attention to the fact that spinal arachnoid membranes may be a potential reservoir for Toxoplasma gondii. SUMMARY OF BACKGROUND DATA: Central nervous system toxoplasmosis is a common manifestation in patients who are immunodeficient. Reports on the spinal toxoplasmosis are rare and focused on spinal cord involvement. METHODS: An adult patient presented with symptoms of spastic paraparesis that had begun 13 years before admission. Thoracic spinal magnetic resonance imaging showed small lesions in posterior subarachnoid space at Th7-Th8. A Th7-Th8 laminectomy was performed. Intradural-extramedullary lesions were excised. RESULTS: Clinical, immunologic, and pathologic examinations showed adhesive spinal arachnoiditis associated with osteoid formation caused by past toxoplasmic infection. There was no impairment of the immunologic defense system. CONCLUSION: Where no causative factor is found in serious spinal adhesive arachnoiditis, the possibility of spinal toxoplasmosis should also be investigated. PMID- 11474363 TI - Sacral radiculopathy secondary to multicentric osteosarcoma. AB - STUDY DESIGN: A case of multicentric osteosarcoma presenting with sacral radiculopathy is reported. OBJECTIVE: To present unusual clinical and radiologic findings of multicentric osteosarcoma. SUMMARY OF BACKGROUND DATA: Multicentric osteosarcoma is a rare variant of osteosarcoma. To the authors' knowledge, no cases of multicentric osteosarcoma presenting as sacral radiculopathy have been reported previously. METHODS: A 14-year-old boy had a large sacral tumor extending into the spinal canal, which was found to account for the initial symptoms, which mimicked those of herniated nucleus pulposus. At diagnosis, a bone survey showed multiple foci of osteosarcoma in the long bones. RESULTS: The patient was treated with chemotherapy, but died of the disease 8 months after the initial presentation. CONCLUSION: Multicentric osteosarcoma should be considered in the differential diagnosis for a pediatric patient with low back pain and sciatica. PMID- 11474365 TI - An immunohistochemical study of the dorsal capsule of the lumbar and thoracic facet joints. AB - STUDY DESIGN: The molecular composition of the extracellular matrix in the dorsal capsules of lumbar and thoracic facet joints was analyzed immunohistochemically. OBJECTIVES: To determine whether the immunohistochemical profile of the lumbar joint capsule suggests a role of the capsule in limiting axial rotation of the lumbar motion segment. SUMMARY OF BACKGROUND DATA: During axial rotation of the lumbar vertebrae, the axis of rotation shifts toward the facet joints in the direction of rotation. Thus, the capsule of the opposing joint should become tensed and wrap around the inferior articular process. Previous studies suggest that wrap-around ligaments are fibrocartilaginous. However, thoracic joint capsules are largely shielded from such loading and should be purely fibrous. METHODS: Dorsal capsules were removed from lumbar and thoracic facet joints of six adult cadavers. Specimens were immunolabeled with monoclonal antibodies for collagens, chondroitin, dermatan and keratan sulfates, versican, tenascin, aggrecan and link protein. Antibody binding was detected using the Vectastain ABC 'Elite' peroxidase kit (Vector Laboratories, Inc., Burlingame, CA). RESULTS: Both lumbar and thoracic joint capsules immunolabelled for most glycosaminoglycans and for Type I, III and VI collagens. However, labeling for Type II collagen, chondroitin-6-sulfate, aggrecan, and link protein was restricted to lumbar capsules. Such labeling was constantly seen at entheses and occasionally in the midsubstance. CONCLUSIONS: The molecular composition of the lumbar joint capsule suggests that it acts as a fibrocartilaginous, 'wrap-around' ligament that withstands compression in addition to tension during torsional movements of the lumbar spine. It wraps around the inferior articular process as rotation occurs and limits further movement. PMID- 11474366 TI - Cross-cultural adaptation of the lumbar North American Spine Society questionnaire for Italian-speaking patients with lumbar spinal disease. AB - STUDY DESIGN: A cross-cultural adaptation and cross-sectional study of a sample of lumbar spine patients, with a subsample followed prospectively for retest reliability. OBJECTIVES: To assess the Italian version instrument reliability and validity. SUMMARY OF BACKGROUND DATA: The orthopaedic outcome measurements have been usually focused on objective parameters as radiograph measures or other technical aspects. However, these parameters are weakly related with outcomes that are more relevant to patients as functional status and symptoms. In the last ten years, the patient-oriented measures have become an important aspect of spinal clinical outcome evaluation. The most common instruments to assess patient perspective are self-administered questionnaires that must be validated by a widely accepted process to evaluate reliability and validity, which are fundamental for every instrumental measure. METHODS: The North American Spine Society (NASS) questionnaire was culturally adapted for Italian-speaking people following the Guillemin criteria. The Italian version was tested on 74 consecutive patients who were referred to the authors' department and suffered from low back pain with leg irradiation. The results were compared with other validated patient-oriented measures. Forty-eight-hour retests were performed on a subsample of 45 patients. RESULTS: The questionnaire was favorably accepted by patients. The lumbar spine pain and disability and neurogenic symptoms subscales showed a high correlation with other patient-oriented measures, as hypothesized, and it also showed good values on test-retest. CONCLUSIONS: The questionnaire should be considered for patient health status monitoring and for clinical trials. PMID- 11474367 TI - Implementation barriers for general practice guidelines on low back pain a qualitative study. AB - STUDY DESIGN: Qualitative study design, using semi-structured interviews. OBJECTIVE: To explore factors that determine non-adherence to the guidelines for low back pain. SUMMARY OF BACKGROUND DATA: Guidelines for low back pain have been published in the past decade in various countries. In the Netherlands, general practitioners adhere to them to a fair extent, and it is unclear whether room for improvement remains. METHODS: Forty semistructured, in-depth interviews were conducted with twenty patients who consulted for low back pain, and with their general practitioners. The interviews were fully transcribed and analyzed qualitatively. RESULTS: Patients often had limited expectations of the consultation. They wanted to hear a diagnosis or expected to receive simple advice. The general practitioners said they were well informed about the guideline and mostly agreed with its content. Reasons for non-adherence were mainly related to patients' experiences in the past and general practitioners' interpretations of their preferences. General practitioners stated that they were inclined to give in to patients' demands, for example the request for radiographic films or a referral to a physical therapist. In general, patients and their general practitioners were satisfied with the chosen management. CONCLUSIONS: Improvement of the quality of back pain care may still be possible. Implementation strategies should aim at training physicians in communication skills, especially about subjects for debate, where patients' beliefs and experiences color their expectations. PMID- 11474369 TI - Clinical and laboratory findings of cytomegalovirus infection in 115 hospitalized non-immunocompromised adults. AB - We report a retrospective study of 115 hospitalized non-immunocompromised adults with proved or presumed diagnosis of cytomegalovirus infection. Clinical symptoms were fever (95%), constitutive symptoms (80%), joint and muscle pain (41%), shivering (32%), abdominal pain (26%), non-productive cough (20%), cutaneous eruption (20%), and diarrhea (10%). Examination found hepatomegaly (25%), splenomegaly (23%), cutaneous rash (20%), adenopathy (19%), pharyngitis (9%), jaundice (3%) or signs of meningeal irritation (1%). Seventeen patients had a gastrointestinal form (hepatitis, jaundice, colitis, antral gastritis or cholecystitis), eight had a pattern of hemopathy, two interstitial pneumonitis, two pericarditis, two immune thrombocytopenic purpura, two a polymyalgia rheumatica-like pattern, one thrombotic thrombocytopenic purpura, one cutaneous vasculitis and one meningoencephalitis. Sixty-four percent of the patients had atypical lymphocytosis. Hepatocellular injury occurred in 90% of the patients. Nineteen of the patients had biological immune abnormalities. Cytomegalovirus infection should be mainly suspected in any patient with persistent fever, isolated or associated with signs of poor specificity, or in some patients with visceral manifestations of initially unknown origin. PMID- 11474370 TI - [Bacterial spondylodiskitis: pyogenic or tuberculous?]. AB - We retrospectively studied the epidemiological, clinical, radiological and evolutive aspects of bacterial spondylodiskitis observed in 39 patients diagnosed from January 1989 to December 1998, in order to find criteria to distinguish between pyogenic spondylodiskitis (PS) and tuberculous spondylodiskitis (TS). During the study period, we collected 13 patients (33.3%) with PS and 26 patients (66.7%) with TS. The mean age was 44 years. The delay in diagnosis was much longer in TS (8.4 months) than in PS (2 months). Diagnosis was certain in 6 cases (46%) of PS and in 12 cases (46%) of TS. Clinically, paravertebral abscesses and neurologic complications were significantly more frequent in TS. The main causative agents in PS were staphylococci. Diagnosis of spondylodiskitis is based on the imaging techniques, especially magnetic resonance imaging which is more sensitive and specific than computed tomography. Except in patients with positive blood cultures or positive urine culture in PS, and isolation of Mycobacterium tuberculosis in another visceral foci in TS, bacteriologic diagnosis was based on discovertebral needle biopsy. The yield of this technique was poor in our study (12.5%), even if bacteriologic confirmation of spondylodiskitis was made in 15 patients (38.5%). In the other cases, diagnosis rested on a set of clinical, biological and radiological criteria. PMID- 11474371 TI - [From rare to frequent diseases: illustration from digestive tract oncology]. PMID- 11474372 TI - [Digestive tract oncologic genetics: its role in the crossroads between hepato gastroenterology and medical genetics]. PMID- 11474373 TI - [Genetic diagnosis of cancers? Which cancers should be screened?]. AB - Our knowledge in the genetic basis of hereditary human cancer has improved over the last 10 years. Molecular diagnoses have become feasible in major hereditary cancer-prone syndromes, as well as in other hereditary diseases in which cancer appears as symptoms or complications. The major cancer-prone syndromes are described here with a table summarizing the links between various cancer types and the genetic syndromes to which they could belong. Clinicians should be aware of these new diagnostic tools. Specific consultations in oncogenetics are being settled to help clinicians and patients in the course of establishing molecular diagnoses, and providing guidelines for a better surveillance of high-risk persons. PMID- 11474374 TI - [Practical data on digestive tract oncogenetics]. AB - Inherited conditions predisposing to cancers have long been recognized. Specific medical recommendations have more recently been proposed to avoid the development of advanced cancers in such high-risk families. The identification of the first genes involved in monogenic dominant cancer-prone conditions have opened the ways to their predictive diagnosis in asymptomatic members in at-risk families. The development of such procedures has required specific involvement for clinicians and molecular geneticians. The example of familial predispositions to colonic cancers has been chosen to illustrate the practical issues, and to help the physicians in charge of at-risk families to offer their patients an access to these emerging possibilities in cancer prevention. A table summarizes the practical issues requested to refer a family and initiate the procedure of molecular testing. PMID- 11474375 TI - [Intestinal polyposis]. PMID- 11474376 TI - [Rapidly progressive dementia disclosing primary angiitis of the central nervous system]. AB - Angiitis of the central nervous system involves a wide spectrum of clinical symptoms. Complementary exams (CSF, neuroradiological exams) are not very specific and sensitive. Diagnostic procedures often require cortical and leptomeningeal biopsy. Treatment, based on corticosteroids and cyclophosphamid, have greatly improved prognosis. We report a rapidly progress dementia syndrome that revealed angiitis of the central nervous system. Corticosteroid treatment did not prevent a fatal outcome. Autopsy findings confirmed the diagnosis of central nervous leucocytoclastic angiitis. PMID- 11474377 TI - [RS3PE syndrome: an acute edematous polyarthritis of the elderly with variable prognosis]. AB - The RS3PE syndrome or subacute edematous polyarthritis of the elderly remains a doubtful entity. We report three cases that exhibited different courses: complete recovery, definite rheumatoid polyarthritis, and chronicity as a sign of myelodysplasic disease. These three different courses raise the question of whether RS3PE is a disease or a syndrome. Actually, the use of the term RS3PE syndrome should be restricted to cases with a favorable outcome. Definitive diagnosis thus cannot be reached before complete recovery. PMID- 11474378 TI - [Septic arthritis of a lumbar facet joint. A case report]. AB - Septic arthritis of the posterior lumbar joints is extremely rare in comparison with spondylodiscitis which is much more common. We report a case of an 86-year old women with septic arthritis of the left L4-L5 lumbar facet joint associated with endocarditis. Arthritis diagnosis was made on CT scan and MRI, infection by Staphyloccocus aureus was proved by blood cultures. Heart growth was seen by echocardiography. Twenty-three cases were reported in the literature. Clinical and biological data failed to discriminate between facet joint septic arthritis and spondylodicitis. Diagnosis is established on imaging findings, computed tomography and magnetic resonance imaging, completed by blood cultures and, if they are negative, by aspiration-biopsy. Appropriate antimicrobial therapy is usually successful. Some back pain generally persists. In conclusion, lumbar pain with fever without spondylodiscitis is suggestive of septic arthritis of a lumbar facet joint. Epiduritis associated in 60% patients requires rapid treatment. PMID- 11474379 TI - [Sulfasalazine neurotoxicity]. AB - We report a case of seizures with acute encephalopathy in a female patient under sulfasalazine treatment for polyarthritis. Neurotoxicity secondary to sulfasalazine was suspected. This side effect has seldom been reported in the literature. PMID- 11474380 TI - [Common bile duct dilatation: one more complication in patients with autosomal dominant polycystic kidney disease]. PMID- 11474381 TI - Physiologic assessment of coronary artery stenoses. PMID- 11474382 TI - A comparison of 3 types of covered metal stents for the palliation of patients with dysphagia caused by esophagogastric carcinoma: a prospective, randomized study. AB - BACKGROUND: There are currently 3 types of covered metal stents available in Europe for palliation of patients with malignant dysphagia. Their relative merits have not been compared in a prospective, randomized study. METHODS: One hundred consecutive patients with esophagogastric carcinoma were randomized to placement of an Ultraflex stent, a Flamingo Wallstent, or a Gianturco-Z stent. Malignant strictures of the esophagus were treated by insertion of a small-diameter stent (n = 71), whereas those involving the gastric cardia were treated with a large diameter stent (n = 29). RESULTS: At 4 weeks, dysphagia had improved in all patient groups (p < 0.001), but the degree of improvement did not differ among the 3 groups (p = 0.14). There were differences among the 3 stent types with respect to major complications (Ultraflex stent: 8/34 [24%], Flamingo Wallstent: 6/33 [18%], and Gianturco-Z stent: 12/33 [36%]), but these were not statistically significant (p = 0.23). Nine patients (26%) with an Ultraflex stent, 11 (33%) with a Flamingo Wallstent, and 8 (24%) with a Gianturco-Z stent had recurrent dysphagia (p = 0.73), mainly because of tumor overgrowth or stent migration; 12 of 13 episodes of migration involved small-diameter stents in the esophagus. CONCLUSIONS: All 3 covered metal stents evaluated offer the same degree of palliation of patients with malignant dysphagia. Placement of Gianturco-Z stents was associated with more complications as compared with Ultraflex stents and Flamingo Wallstents. Although stent migration is reduced by increasing stent diameter, tumor overgrowth remains an intractable problem that requires a new approach. PMID- 11474383 TI - Long-term follow-up after biliary stent placement for postoperative bile duct stenosis. AB - BACKGROUND: The outcome of temporary biliary stent placement for postoperative bile duct stenosis was retrospectively evaluated with the main aim of assessing long-term complications after stent removal. METHODS: ERCP was performed between 1981 and 1991 in 74 patients with postoperative bile duct stenoses. Two 10F stents were inserted for a maximum of 12 months with stent exchange every 3 months to avoid cholangitis caused by clogging. RESULTS: Stent insertion failed in 11 patients with complete and 4 patients with incomplete biliary obstruction. Early complications occurred in 14 patients (19%) including 2 deaths. Therefore 57 patients were included in the stent phase of the study. In 10 patients the referring physician did not adhere to the treatment protocol, and nonelective stent exchange for jaundice and/or cholangitis was necessary in 7 (70%). Of the 47 patients treated according to protocol, complications developed in 40% during the period with stents in situ. Stents were eventually removed in 44 patients who were subsequently followed for a median of 9.1 years. Late complications developed in 15 patients (34%) including recurrent stenosis in 9 (20%). All cases of recurrent stenosis occurred within 2 years of stent removal. CONCLUSIONS: Endoscopic treatment is feasible in 80% of patients who undergo an ERCP for postoperative bile duct stenosis. After stent insertion and during the time with stents in situ, complications occur at a significant rate but are usually mild or reflect the patient's underlying condition. After stent removal, recurrent stenosis develops in 20% of patients within 2 years of stent removal. Endoscopic treatment should be the initial management of choice for postoperative bile duct stenosis. PMID- 11474384 TI - Long-term results of endoscopic management of postoperative bile duct strictures with increasing numbers of stents. AB - BACKGROUND: Endoscopic dilation with stents has been proposed as an alternative to hepaticojejunostomy for management of postoperative biliary strictures. Good long-term results with double 10F plastic stent insertion for 1 year have been reported in 74% to 90% of cases. This is a review of our experience with a more aggressive approach. METHODS: The technique, short-term results, and long-term results of placement of increasing numbers of stents until complete disappearance of the biliary stricture are reported. At each exchange, the maximum possible number of stents in relation to the tightness of the stricture and diameter of the bile duct were inserted. All stents were removed at the end of treatment. RESULTS: The records of 45 of 55 patients with postoperative biliary strictures treated in this manner and observed consecutively were reviewed retrospectively. By intention-to-treat analysis the success rate was 89% (40/45). Early complications developed in 4 (9%) patients (3 cholangitis, 1 pancreatitis) and stent occlusion that required early exchange occurred in 8 (18%) patients. There was 1 death caused by a stroke 2 months after a stent exchange. Forty-two patients completed the protocol (mean number of stents 3.2 +/- 1.3; range 1-6). Mean duration of treatment was 12.1 +/- 5.3 months (range 2-24 months). Two patients died of unrelated causes during follow-up. Among the remaining 40 patients there was no recurrence of symptoms caused by relapsing biliary stricture at a mean follow-up of 48.8 months (range 2-11.3 years). One patient sustained 2 episodes of cholangitis but without stricture recurrence. CONCLUSIONS: This more aggressive approach to endoscopic treatment with stents may improve long-term results for patients with postoperative biliary strictures. PMID- 11474385 TI - Routine versus "on demand" sedation and analgesia for colonoscopy: a prospective randomized controlled trial. AB - BACKGROUND: The safety and tolerance of routine sedation and analgesia versus "on demand" sedation were compared in patients undergoing colonoscopy. METHODS: Two hundred forty-nine outpatients were randomly assigned to one of two groups. Group A (n = 125) received midazolam, 0.07 mg/kg intravenously plus meperidine, 0.77 mg/kg intravenously immediately preceding the colonoscope insertion. Group B (n = 124) received the same medication upon request during the procedure. Tolerance was assessed 24 hours later by phone interview performed by a nurse blinded to the medication regimen administered. RESULTS: Eighty-three patients (66%) in Group B required sedation during colonoscopy. Among men in Group B more than 60 years of age, only 23% required sedation. The proportion of patients reporting moderate or severe pain (34% vs. 12.1%, p < 0.001) and of those stating they would not be willing to undergo colonoscopy again in the future (22% vs. 9.7%, p < 0.005) was significantly higher in the "on demand" sedation group. By multivariate analysis the randomization group was the single variable independently associated with both such outcomes. The frequency of side effects was similar in the two groups. CONCLUSIONS: Administration of sedative and analgesic drugs routinely before colonoscopy is superior to "on demand" sedation in terms of tolerance and is not associated with an increase in side effects. PMID- 11474386 TI - Diagnostic yield of open access colonoscopy according to appropriateness. AB - BACKGROUND: Open-access endoscopy allows physicians to directly schedule endoscopic procedures for their patients without prior consultation. Evaluation of both appropriateness and diagnostic yield of endoscopic procedures is critical when assessing the costs and benefits of endoscopy in an open-access setting. The aim of this study was to assess the appropriate use of colonoscopy in an open access system and to establish the yield of diagnostic information relevant to patient care. METHODS: Overall, 1123 consecutive patients referred for open access colonoscopy were prospectively enrolled in the study. The American Society for Gastrointestinal Endoscopy (ASGE) guidelines were used to assess the relationship between the appropriate use of colonoscopy and the presence of relevant endoscopic findings. RESULTS: The rate of colonoscopies "generally not indicated" according to ASGE guidelines was 29% (39% for primary care physicians and 23% for specialists; p < 0.0001). A relevant endoscopic finding was detected in 338 examinations (35%). The diagnostic yield was significantly higher for "generally indicated" colonoscopies (43%) compared with "generally not indicated" procedures (16%) (p < 0.001). CONCLUSIONS: Although the rate of inappropriate use of colonoscopy was high, open-access colonoscopy was effective in detecting neoplastic lesions. Because most of these were detected during examinations performed for appropriate indications, the appropriateness of the indication emerges as crucial to the cost-effectiveness of an open-access system. PMID- 11474387 TI - Cisapride does not improve precolonoscopy bowel preparation with either sodium phosphate or polyethylene glycol electrolyte lavage. AB - BACKGROUND: Oral sodium phosphate solution (NAP) and polyethylene glycol electrolyte lavage (PEG-EL) are used for precolonoscopy bowel preparation. The benefit of adding cisapride to PEG-EL is controversial, and its influence on the effectiveness of NAP has not been investigated. The primary aim of this study was to determine whether cisapride improves the effectiveness and/or tolerableness of bowel preparation with either NAP or PEG-EL. METHODS: In 187 patients undergoing colonoscopy, a randomized, double-blind, placebo-controlled trial with a Latin square design was conducted to compare 4 different bowel preparations: NAP plus either cisapride (10 mg; 2 doses) or placebo, or PEG-EL plus either cisapride (10 mg; 1 dose) or placebo. Quality of the bowel preparation was graded by the endoscopist according to the amount of stool present in the colon (excellent, satisfactory, unsatisfactory). To assess tolerability, patients rated 8 symptoms, the taste of the lavage solution, and the ease of preparation on a 5-point scale (1: easy; 5: distressing). RESULTS: Endoscopists scored the bowel preparation as either excellent or satisfactory as follows: NAP: cisapride 50% versus placebo 61% (p = 0.3); PEG-EL: cisapride 80% versus placebo 78% (p = 1.0). Cisapride did not improve tolerability or the frequency of adverse symptoms associated with either solution. The ease of bowel preparation was significantly better in the NAP group versus PEG-EL group (mean score 1.8 versus 2.8; p < 0.0001). CONCLUSIONS: Cisapride does not improve the quality of bowel preparation with either NAP or PEG-EL. NAP is better tolerated by patients than PEG-EL; however, PEG-EL results in better bowel preparation. PMID- 11474388 TI - Novel adaptation of brush cytology technique for short-term primary culture of squamous and Barrett's esophageal cells. AB - BACKGROUND: Esophageal disease research is impeded by a paucity of nonmalignant cell lines. Furthermore, culture of primary esophageal cells is difficult because of a lack of cell adhesion and contamination. The aim of this study was to develop a short-term culture method to facilitate cell physiologic studies by using primary esophageal cells. METHODS: By using a cytology brush, squamous and Barrett's epithelial cells were obtained from the esophagus of patients undergoing upper endoscopy. Cells were brushed onto chamber slides and allowed to adhere to the surface. Primary culture media was then added and cells were maintained at 37 degrees C for up to 72 hours. Cell yield and viability were calculated after trypan blue staining. For cell physiologic studies, cells were loaded with pH-sensitive dye BCECF-AM and intracellular pH measured on a dual excitation fluorescence microscope after an ammonium chloride prepulse. At the end of the physiologic experiments, cells were fixed in methanol and acetone and the cell type was verified with cytokeratin immunocytochemistry. RESULTS: Viable human esophageal cells were maintained in culture for up to 72 hours. The cells extruded trypan blue, and BCECF-AM was cleaved to BCECF by an intact cell membrane and permitted intracellular pH measurements. The epithelial cell origin of the cells was confirmed by cytokeratin staining. There was no contamination over the culture period and no overgrowth by lymphocytes or fibroblasts. CONCLUSIONS: This novel adaptation of brush cytology technique enables short-term culture of esophageal cells for in vitro studies. This rapid, simple primary culture technique is suitable for endoscopy and does not require the immediate, time-consuming laboratory techniques. PMID- 11474389 TI - Metachronous squamous cell carcinoma of the esophagus arising after endoscopic mucosal resection. AB - BACKGROUND: Endoscopic mucosal resection (EMR) is being used increasingly to treat early stage esophageal carcinoma. However, the preserved esophageal mucosa may be the source of new lesions. The aims of this study were to analyze the frequency of metachronous esophageal carcinoma after EMR and to determine whether minute iodine unstained areas often associated with squamous cell carcinoma develop into carcinoma. METHODS: Eighty-two patients with esophageal squamous cell carcinoma who underwent EMR were studied. Based on the iodine staining pattern at initial EMR, they were divided into those with uniform (group U) and scattered (group S) types of background mucosa. Patients were followed by endoscopy with iodine staining (group U: median 39 months, range 12 to 71 months; group S: median 38 months, range 14 to 68 months). RESULTS: In total, 12 (14.6%) of 82 patients were found to have metachronous esophageal carcinoma during follow up, including 6 (37.5%) of 16 patients in group S. The cumulative proportion of metachronous carcinoma-free subjects was significantly lower in group S than group U (p = 0.0048). CONCLUSIONS: Primary esophageal carcinoma develops frequently in patients who have undergone EMR for esophageal squamous carcinoma. The high frequency of metachronous carcinoma may be attributed to field carcinogenesis. Careful long-term endoscopic observation is required for patients who undergo EMR for esophageal carcinoma, especially those with scattered-type iodine staining of the background mucosa. PMID- 11474390 TI - Light-induced autofluorescence spectroscopy for the endoscopic detection of esophageal cancer. AB - BACKGROUND: Any innovative optical system that facilitates the early endoscopic detection of neoplastic change in the GI mucosa has the potential to greatly improve survival and quality of life for patients prone to have GI malignancies develop. The present article describes light-induced autofluorescence spectroscopy with violet-blue excitation light for in vivo diagnosis of cancerous tissue of the esophagus during routine endoscopy. METHODS: One hundred twenty nine endogenous fluorescence spectra were obtained from normal mucosa and malignant lesions in 9 patients with squamous cell cancer and 4 with adenocarcinoma of the esophagus. Following spectrographic measurements, biopsy specimens were obtained for definitive classification of the spectra. A special light source capable of delivering either white or violet-blue light for excitation of tissue autofluorescence by means of an endoscope was used. Endogenous fluorescence spectra emitted by tissues were detected with a fiberoptic probe and analyzed with a spectrograph. RESULTS: Squamous cell cancer and adenocarcinoma of the esophagus exhibit specific changes in the emitted fluorescence spectra as compared with normal mucosa. Based on the results obtained in earlier studies, malignant and benign spectra were differentiated with the aid of a mathematical algorithm. By using this algorithm, a sensitivity of 97% and specificity of 95% were obtained for the diagnosis of esophageal carcinoma. CONCLUSIONS: Light-induced fluorescence spectroscopy is useful for the endoscopic detection of squamous cell cancer and adenocarcinoma of the esophagus. This spectroscopic study provides a basis for the design of a simplified autofluorescence imaging system for detection of esophageal neoplasms. PMID- 11474391 TI - Endoscopic management of tumors of the major duodenal papilla: Refined techniques to improve outcome and avoid complications. AB - BACKGROUND: Adenomas of the major duodenal papilla have malignant potential and are traditionally treated by pancreaticoduodenectomy. This is a report of our experience with endoscopic management and a description of techniques for decreasing complications and enhancing efficacy. METHODS: Forty-one patients were referred for endoscopic management of papillary tumors. If there was no duct invasion and the appearance suggested a benign lesion, biductal sphincterotomy with pancreatic duct stent placement was performed. If the lesion could be elevated by injection of an epinephrine solution, piecemeal resection was performed. The base of the lesion was thermally ablated as needed. Resection/ablation together with stent removal was performed 1 month later. RESULTS: Nine patients (22%) had lesions other than papillary adenoma or cancer. Malignant appearance, ductal stricturing, or extension into the ducts was found in 16 of 41 patients (39%) in whom biopsy specimens alone were obtained. Three patients with adenomas (7%) did not undergo endoscopic resection (because of extremely large lesions and/or comorbid illnesses). Thirteen patients with adenomas (32%) had endoscopic resection; 12 (92%) were lesion-free after 32 ERCPs (mean 2.7). Endoscopic management was unsuccessful in 1 patient (8%). Pancreatitis developed in 1 patient. CONCLUSIONS: Endoscopically treatable papillary neoplasms can be identified on the basis of endoscopic, radiographic, and biopsy features. Preresection sphincterotomy, stent placement, elevation by epinephrine injection, and piecemeal resection may reduce complications and permit more aggressive treatment. PMID- 11474392 TI - Stent placement in the pancreatic duct prevents pancreatitis after endoscopic sphincter dilation for removal of bile duct stones. AB - BACKGROUND: Pancreatitis is the most serious complication of endoscopic sphincter dilation. The aim of this study was to determine whether temporary stent placement in the main pancreatic duct decreases the frequency of pancreatitis and level of hyperamylasemia. METHODS: Stents were placed in the pancreatic duct after endoscopic sphincter dilation in 40 consecutive patients with bile duct stones. Stents were removed endoscopically 3 days later. Changes in serum amylase and the frequency of pancreatitis for this group were compared with those in 92 patients who underwent sphincter dilation without pancreatic duct stent placement. RESULTS: Stent placement was successful in 38 of 40 patients. Although the difference in the frequency of pancreatitis was not significantly different between stent and control groups, there was a trend toward a decrease in pancreatitis in the stent group. The level of postprocedure hyperamylasemia was significantly less in the stent group (p < 0.05). There were no procedure-related complications. CONCLUSIONS: Temporary placement of a stent in the pancreatic duct after sphincter dilation for removal of bile duct stones has a beneficial effect in terms of postprocedure hyperamylasemia and appears to reduce the frequency of postprocedure pancreatitis. PMID- 11474393 TI - Bacteremia after endoscopic injection of N-butyl-2-cyanoacrylate for gastric variceal bleeding. AB - BACKGROUND: Cyanoacrylate may form a barrier that prevents bacterial invasion when used in tissue. Because cyanoacrylate polymerizes within seconds on contact with aqueous media, it is used worldwide to arrest gastric variceal bleeding. The aim of this study was to determine the frequency of bacteremia after endoscopic cyanoacrylate injection for gastric variceal bleeding. METHODS: Patients with cirrhosis who underwent endoscopic cyanoacrylate injection for gastric variceal bleeding were included. Patients with cirrhosis who underwent upper endoscopy for nonvariceal upper GI bleeding were recruited as controls. Patients with infection before endoscopy were excluded. Blood was cultured in both groups. Injection needles and endoscope accessory channels were cultured in the cyanoacrylate injection group. RESULTS: More patients injected with cyanoacrylate had positive blood cultures in comparison with the control group (15/47 vs. 1/47, p < 0.0001). In the cyanoacrylate injection group, the volume of blood transfused and Child Pugh score were factors associated with the occurrence of bacteremia. Most episodes of bacteremia were transient, except for 1 patient who died of sepsis. Most of the microorganisms cultured from blood samples were identical to those cultured from injection needles (65%) and accessory channels (90%). CONCLUSIONS: Endoscopic cyanoacrylate injection for gastric varices does not limit the spread of bacteria. The endoscope accessory channel was the major source of bacteria. Most episodes of bacteremia were transient and uneventful. PMID- 11474394 TI - High-resolution endoscopic imaging of the GI tract: a comparative study of optical coherence tomography versus high-frequency catheter probe EUS. AB - BACKGROUND: Both optical coherence tomography (OCT) and catheter probe EUS (CPEUS) are candidates for high-resolution imaging of the GI wall, but their potential roles in this clinical context have not been investigated. METHODS: OCT and CPEUS were used to image normal-appearing portions of the GI tract at the same sites. CPEUS was performed with a 20-MHz or a new 30-MHz catheter probe. RESULTS: Forty-four histologically confirmed normal sites in 27 patients were evaluated. With OCT, mucosa and muscularis mucosa were clearly seen at all sites. Except for stomach, OCT demonstrated the submucosa in all sites. OCT penetration ranged from 0.7 to 0.9 mm. Microscopic structures such as esophageal glands, intestinal villi, colonic crypts, and blood vessels were easily identified. CPEUS penetration ranged from 10 mm to 20 mm, and 5 to 7 distinct layers were discernible. However, both mucosa and submucosa were seen as thin layers without microscopic detail. CONCLUSION: OCT resolution is superior to high-frequency CPEUS, but depth of penetration is limited to mucosa and submucosa. OCT images the major structural components of the mucosa and submucosa whereas CPEUS does not. Potentially, OCT and high-frequency CPEUS may be complementary for clinical imaging. PMID- 11474395 TI - Acute necrotizing esophagitis. PMID- 11474396 TI - Villous adenoma of the common hepatic duct. PMID- 11474397 TI - Choledochocolonic fistula secondary to primary choledocholithiasis. PMID- 11474398 TI - Duodenal metastasis from lung cancer presenting as obstructive jaundice. PMID- 11474399 TI - Usefulness of Dennis Colorectal Tube in endoscopic decompression of acute, malignant colonic obstruction. AB - BACKGROUND: Colonoscopic decompression has been attempted in patients with acute colonic obstruction caused by colon cancer to avoid emergency surgery and multiple subsequent operations but has usually been unsuccessful. This is an evaluation of the usefulness of a new device for endoscopic decompression in these patients. METHODS: Nine consecutive patients (6 men, 3 women; 65 to 89 years of age) with acute colonic obstruction resulting from colon cancer underwent endoscopic decompression with a Dennis Colorectal Tube. RESULTS: In all 9 patients (1 with carcinoma of the sigmoid colon, 3 with carcinoma of descending colon, 2 with carcinoma of the transverse colon, 2 with carcinoma of the ascending colon, and 1 with carcinoma of the cecum), endoscopic decompression was successful. After decompressing and cleansing the colon for several days, a one stage operation was possible in all patients. CONCLUSION: Dennis Colorectal Tube is an excellent device for endoscopic decompression of acute, malignant colonic obstruction. PMID- 11474400 TI - The role of percutaneous transhepatic papillary balloon dilation in percutaneous choledochoscopic lithotomy. AB - BACKGROUND: When choledochoscopic lithotomy with basket and electrohydraulic lithotripsy is used to remove intrahepatic duct stones, fragments or small stones usually remain in the bile duct that are too small to be captured with a basket. METHODS: An attempt was made to remove stone fragments in 16 patients with intrahepatic duct stones by antegrade balloon dilation of the sphincter of Oddi with a conventional balloon catheter. After balloon dilation, remnant stones and sludge were pushed through the papilla with the choledochoscope. RESULTS: Bile duct stones were completely removed in 12 of 16 patients (75%); stones were removed in 1 session. There was no clinical evidence of procedure-related pancreatitis or fatal complications. CONCLUSIONS: Percutaneous transhepatic papillary balloon dilation of the sphincter of Oddi and clearance of remnant bile duct stones and stone fragments with the tip of choledochoscope is simple and effective in patients undergoing percutaneous transhepatic choledochoscopic lithotomy. PMID- 11474401 TI - Placement of PEG tubes through previously placed self-expanding esophageal metal stents. AB - BACKGROUND: Self-expanding metal stents are frequently used to palliate patients with malignant dysphagia and close tracheoesophageal fistulae. Despite proper stent positioning and deployment, in a subset of patients there is no improvement in dysphagia, closure of tracheoesophageal fistulae, or resolution of anorexia. Such patients may require a PEG tube. It has been suggested that PEG placement through a preexisting esophageal stent is problematic because of the risks of gastrostomy tube impaction within the stent and resultant stent migration. METHODS: Case records were retrospectively reviewed of 9 consecutive patients with indwelling esophageal self-expanding metal stents undergoing attempted PEG. OBSERVATIONS: PEG tube placement was successful in all patients. In 1 patient, the stent migrated distally into the stomach during PEG placement. This was managed endoscopically without further complication. CONCLUSIONS: PEG placement in patients with previously placed esophageal self-expanding metal stents is a relatively safe and feasible procedure, although stent migration may occur. PMID- 11474402 TI - Pancreatic clonorchiasis diagnosed by EUS-guided pancreatic duct aspiration. PMID- 11474403 TI - Colonoscopic clipping of fecal fistula that occurred as a postoperative complication in patients with perforated appendicitis: two case reports. PMID- 11474404 TI - Endoscopic closure of a colovesical fistula. PMID- 11474405 TI - Clinical and endoscopic improvement of lymphocytic gastritis with eradication of Helicobacter pylori. PMID- 11474406 TI - Tension pneumothorax complicating ERCP in a patient with a Billroth II gastrectomy. PMID- 11474407 TI - Esophagitis dessicans superficialis with bulla in chronic renal failure: a case report. PMID- 11474408 TI - Iatrogenic perforation of the colon during diagnostic colonoscopy: endoscopic treatment with clips. PMID- 11474409 TI - Intramural cyst of the bile duct demonstrated by cholangioscopy and intraductal US. PMID- 11474410 TI - Failure of ERCP to identify biliary leak after laparoscopic cholecystectomy. PMID- 11474411 TI - Endoscopic removal of a phytobezoar obstructing the distal small bowel. PMID- 11474412 TI - Ileal carcinoid tumor with a rose-shaped appearance by dye spraying: diagnosis at colonoscopy. PMID- 11474413 TI - A practical guide for choosing an expandable metal stent for GI malignancies: is a stent by any other name still a stent? PMID- 11474414 TI - Long-term follow-up after biliary stent placement for postoperative bile duct stenosis. PMID- 11474415 TI - GI endoscopy in Lebanon: past, present, and future. PMID- 11474416 TI - Endoscopic markers in celiac disease: are they useful? PMID- 11474417 TI - Distal rectal tumors are always closer (to the anus) than they seem. PMID- 11474418 TI - Substantial surveillance bias in an unblinded study of pain and adverse effects associated with nitrous oxide inhalation or standard medication for colonoscopy. PMID- 11474419 TI - Cost minimization analysis of alternative strategies for initial staging of esophageal cancer. PMID- 11474420 TI - The analysis of clinical studies: the use of transformations to achieve normality. PMID- 11474421 TI - Hu 210: a potent tool for investigations of the cannabinoid system. AB - The synthetic compound HU 210 displays a multiplicity of biochemical, pharmacological, and behavioral effects, most of which have been demonstrated to be dependent on a selective agonistic activity at CB(1) and CB(2) cannabinoid receptors and to involve the main neurotransmitter systems. Results obtained in various studies suggest a potential clinical application of this highly potent drug (e.g., as antipyretic, antiinflammatory, analgesic, antiemetic, and antipsychotic agent) as well as its usefulness in research aimed to develop a better understanding of the involvement of the endogenous cannabinoid system in a number of physiopathological functions. PMID- 11474422 TI - BTS 72664-- a novel CNS drug with potential anticonvulsant, neuroprotective, and antimigraine properties. AB - BTS 72664, (R)-7-[1-(4-chlorophenoxy)]ethyl]-1,2,4-triazolo(1,5-alpha)pyrimidine, was identified as a drug development candidate from a research program designed to discover novel, broad-spectrum, non-sedative anticonvulsant drugs. BTS 72664 antagonized bicuculline (BIC)- and maximal electroshock (MES)-induced convulsions with ED(50) values of 1.9 and 47.5 mg/kg p.o., respectively. In rodents, it has a wide spectrum of activity preventing seizures induced by picrotoxin, pentylenetetrazol, i.c.v. 4-aminopyridine or NMDA, and audiogenic seizures in DBA 2 mice and GEPR-9 rats. BTS 72664 was also effective in preventing convulsions in amygdala-kindled rats The lack of sedative potential was predicted on the basis of wide separation between ED(50) in anticonvulsant models and TD(50) for motor impairment in mice in rotating rod and inverted horizontal grid tests. BTS 72664 is likely to produce its anticonvulsant effect by enhancing chloride currents through picrotoxin-sensitive chloride channels, and by weak inhibition of Na(+) and NMDA channels. It does not act, however, at the benzodiazepine binding site. In addition to its potential use in the treatment of epilepsy BTS 72664 may be useful in the treatment of stroke. At 50 mg/kg p.o. x 4, given to rats at 12 hourly intervals, starting at 15 min after permanent occlusion of middle cerebral artery (MCA), it reduced cerebral infarct size by 31% (measured at 2 days after insult) and accelerated recovery in a functional behavioral model. BTS 72664 prevented increases in extraneuronal concentrations of glutamate, glycine and serine brain levels induced by a cortical insult to rats (cf. cortical spreading depression). It may, therefore, have also antimigraine activity. PMID- 11474423 TI - Gacyclidine: a new neuroprotective agent acting at the N-methyl-D-aspartate receptor. AB - Gacyclidine is a new phencyclidine derivative with neuroprotective properties. Tritiated gacyclidine and its enantiomers bind to NMDA receptors with binding parameters similar to those of other non-competitive NMDA receptor antagonists. The (-)enantiomer, (-)GK11, exhibits an affinity (2.5 nM) similar to that of dizocilpine (MK-801), while the (+)enantiomer, (+)GK11, has a 10 times lower affinity. When its interaction with NMDA receptors is prevented, gacyclidine binds also to "non-NMDA" binding sites which are mainly located in the molecular layer of the cerebellum on the dendritic tree of Purkinje cells. These binding sites do not appear to be related to any known neurotransmitters. In primary cortical cultures, gacyclidine and its enantiomers, at 0.1 to 5.0 microM, prevent glutamate-induced neuronal death. In rats, in vivo neurotoxicity of gacyclidine is far low than that of MK-801. No necrotic neurons were detected in animals sacrificed at 18 or 96 h after treatment with gacyclidine (1, 5, 10 or 20 mg/kg i.v.). At the highest (20 mg/kg) but not the lower doses (1-100 mg/kg) electron microscopy revealed the presence of few cytoplasmic or intramitochondrial vacuoles. In soman-treated monkeys gacyclidine enhanced neuroprotective activity of "three drugs cocktail" (atropine + diazepam + pralidoxime). Moreover, in rats, gacyclidine exerts a dose- and time-dependent neuroprotection in three models of spinal cord lesion. Beneficial effects of gacyclidine include reduction of lesion size and improvement of functional parameters after injury. In traumatic brain injury models gacyclidine improves also behavioral parameters and neuronal survival. Optimal protection is obtained when gacyclidine is administered at 0 to 30 min after injury. It is, therefore, concluded that gacyclidine exhibits neuroprotective effects similar to those of other NMDA receptor antagonists, with the advantage of being substantially less neurotoxic maybe due to its interaction with "non-NMDA" binding sites. PMID- 11474424 TI - Ondansetron: a selective 5-HT(3) receptor antagonist and its applications in CNS related disorders. AB - Ondansetron is a selective 5-hydroxytryptamine(3) (5-HT(3)) receptor antagonist that has been introduced to clinical practice as an antiemetic for cancer treatment-induced and anesthesia-related nausea and vomiting. Its use under these circumstances is both prophylactic and therapeutic. It has a superior efficacy, safety and pharmacoeconomic profile compared with other groups of antiemetics, namely antidopaminergics, antihistamines and anticholinergics. However, its place in the management of anticipatory and delayed vomiting in cancer treatment and as a rescue antiemetic in surgical patients needs to be further explored. Furthermore, recent animal and human research also reflects its possible novel application in the treatment of other disease states, such as alcoholism, cocaine addiction, opioid withdrawal syndrome, anxiety disorders, gastrointestinal motility disorders, Tourette's syndrome and pruritus. This review revisits the widespread physiological and pathological effects of 5-HT and discusses both the basic science literature and the clinical developments responsible for the conventional and novel uses of ondansetron. In addition, new discoveries relating to the effects of ondansetron on other receptors/channels and their possible therapeutic applications are presented. PMID- 11474425 TI - Ion channel diseases of the central nervous system. AB - In the last decade, advances in molecular genetics and cellular electrophysiology have increased our understanding of ion channel function. A number of diseases termed "channelopathies" have been discovered that are caused by ion channel dysfunction. Channelopathies can be caused by autoimmune, iatrogenic, toxic or genetic mechanisms. Mutations in genes encoding ion channel proteins that disrupt channel function are now the most commonly identified cause of channelopathies, perhaps because gene disruption is readily detected by the methods of molecular genetics. Ion channels are abundant in the central nervous system (CNS), but CNS channelopathies are rare; however, they overlap with some important neurological disorders, such as epilepsy, ataxia, migraine, schizophrenia, Alzheimer's disease and other neurodegenerative diseases. It is possible that more CNS channelopathies will be discovered when additional ion channels are characterized and the complex mechanisms of brain function are better understood. At present, increased knowledge of the identity, structure and function of ion channels is facilitating diagnosis and treatment of many channelopathies. PMID- 11474426 TI - Unusual abundance of atypical strains associated with human ocular toxoplasmosis. AB - To facilitate genotyping of Toxoplasma gondii in vitreous fluid of patients with severe or atypical ocular toxoplasmosis, polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP) assays were developed for SAG3 (p43) and SAG4 (p18), 2 single-copy surface antigen genes. Together with strategies for SAG1, SAG2, and B1, multilocus RFLP analyses were performed on PCR amplified parasite DNA present in 12 clinical specimens. Most samples (8/12) were not infected by type II or type III mouse-avirulent strains. Only 1 type III and 3 type II strains were identified, all from immunosuppressed patients. In 6 otherwise healthy adults and in 1 immunosuppressed patient, the SAG1 allele associated with mouse virulence was amplified. Of 12 samples, 3 possessed true type I strains; 5 of 12 had new recombinant genotypes with alleles typical of type I or III strains at all loci examined. The unusual bias toward type I and/or recombinant genotypes bearing the SAG1 type I allele associated with mouse virulence in immunocompetent adults has important implications for the epidemiology and efficacious treatment of ocular toxoplasmosis. PMID- 11474427 TI - Mannose-binding lectin gene polymorphisms as a susceptibility factor for chronic necrotizing pulmonary aspergillosis. AB - It was investigated whether a deficiency of mannose-binding lectin (MBL), which binds Aspergillus species avidly in vitro, could account for chronic necrotizing pulmonary aspergillosis (CNPA), which is seen most commonly in nonimmunocompromised patients. Blood samples were obtained from 11 patients (10 white) with CNPA and were compared with blood samples from 82 white control subjects. MBL haplotype profiles were determined by polymerase chain reaction, using sequence-specific primers and sequence-specific oligonucleotide probing techniques. Seven of the 10 white patients with CNPA had MBL haplotypes that encode for low levels of the protein, compared with 25.6% of the white control subjects (P=.004). Presence of the codon 52 mutation was particularly common in patients with CNPA (P=.015), which suggests a greater involvement of this mutation. PMID- 11474428 TI - Canarypox vaccines induce antigen-specific human gammadelta T cells capable of interferon-gamma production. AB - Induction of human gammadelta T cells was investigated in subjects who were vaccinated with live recombinant canarypox virus expressing human immunodeficiency virus (HIV) proteins or soluble MN rgp120. Both canarypox and rgp120 induced antigen-specific lymphoproliferative and interferon (IFN)-gamma responses. However, only canarypox vaccination induced increased gammadelta T cell responses detectable after secondary in vitro expansion (P<.02). These enhanced gammadelta T cell responses were specific for canarypox but not HIV antigens. Canarypox-specific gammadelta T cells were predominantly Vgamma9(+) and produced intracellular and secreted IFN-gamma. gammadelta T cell lines generated from canarypox vaccinees responded to canarypox antigens but not to mycobacterial antigens shown previously to induce bacille Calmette-Guerin-specific gammadelta T cells. Furthermore, canarypox vaccinations were associated with significantly higher NK cell expansions (P=.02). Increased IFN-gamma production by gammadelta T and NK cells could enhance the induction of protective type 1 memory immunity. Thus, stimulation of gammadelta T cells might be an important feature of live vaccines. PMID- 11474429 TI - National Department of Defense Surveillance for Invasive Streptococcus pneumoniae: antibiotic resistance, serotype distribution, and arbitrarily primed polymerase chain reaction analyses. AB - To provide surveillance among US military personnel and their beneficiaries, 157 invasive Streptococcus pneumoniae clinical isolates were collected systematically from 7 large military hospitals between August 1997 and August 1999. The isolates were studied for antibiotic resistance, and 120 were serotyped and subjected to arbitrarily primed polymerase chain reaction (AP-PCR). Fifty (31.9%) of 157 isolates had intermediate or high-level resistance to penicillin, and 15.9% had multidrug resistance. The most common serotypes were 4, 6B, 9V, 14, 19F, and 23F. Those serotypes associated with penicillin resistance were 6B, 9V, 19A, and 19F. Most invasive disease cases were caused by serotypes included in the currently available 23- and 7-valent pneumococcal vaccines. By use of AP-PCR, 4 DNA groups were correlated with health care site (P< or =.0001). These results are valuable in assessing appropriate use of antibiotics and vaccines against S. pneumoniae in both military personnel and their families. PMID- 11474430 TI - Relationship of Campylobacter toxigenicity in vitro to the development of postinfectious irritable bowel syndrome. AB - Campylobacter enteritis is associated with a significant risk of developing irritable bowel syndrome, but the mechanism is unknown. This study ascertained bowel symptoms in 93 patients 3 months after Campylobacter jejuni enteritis infection. The infecting organisms were cultured, and the effects of culture supernatants on toxin-sensitive epithelial cell monolayers (HEp-2, Green monkey kidney epithelial [Vero], and CHO-K1) were investigated. In all, 50, 43, and 41 of the isolates showed toxigenic effects on HEp-2, CHO-K1, and Vero cells, respectively. Persistently changed bowel habit was reported by 9 of 50 persons with HEp-2 toxin-positive infections but by only 2 of 43 with isolates negative for toxin (chi2=4.0; P<.05). Toxicity to HEp-2 and Vero cells was associated with significantly increased numbers of days with loose stools 6 months after C. jejuni enteritis infection (both, P<.05). Thus, long-term symptoms that occur Campylobacter infection are significantly associated with bacterial toxicity. PMID- 11474431 TI - Role of inducible cyclooxygenase and prostaglandins in Clostridium difficile toxin A-induced secretion and inflammation in an animal model. AB - Cyclooxygenase (Cox)-2 expression and inhibition were investigated in a rabbit ileal loop model of Clostridium difficile colitis and diarrhea. Intestinal tissue stimulated with C. difficile toxin A showed up-regulation of Cox-2 expression in lamina propria macrophages and elevated prostaglandin levels. Toxin A-stimulated loops exhibited severe inflammation and increased secretory volume. Celecoxib, a specific Cox-2 inhibitor, significantly reduced toxin A-induced prostaglandin production. Furthermore, celecoxib (> or =0.02 mg/mL) blocked both histologic damage (mean histologic grade, 1.25 vs. 3.44 in rabbits receiving toxin A alone; P<.0005) and secretion (volume:length ratio, 0.18 vs. 0.72 in those receiving toxin A alone; P=.002) in toxin A-stimulated loops in a dose-related manner. Thus, toxin A induced expression of Cox-2 in the host, and prostaglandins produced through Cox-2 were involved in the mediation of the increased secretion of electrolytes and water and the inflammatory response induced by toxin A. PMID- 11474432 TI - Optochin resistance in Streptococcus pneumoniae: mechanism, significance, and clinical implications. AB - Traditionally, Streptococcus pneumoniae is identified in the laboratory by demonstrating susceptibility to optochin. Between 1992 and 1998, 4 pneumococcal isolates exhibiting optochin resistance were recovered from patients at Children's National Medical Center. Three of the 4 isolates consisted of mixed populations of optochin-resistant and -susceptible organisms. Both subpopulations had identical antibiograms, serotypes, and restriction fragment profiles. The other isolate was uniformly resistant to optochin. Resistant strains had MICs of optochin 4-30-fold higher than susceptible strains, belonged to different serotypes, and had dissimilar restriction fragment profiles, indicating clonal unrelatedness. Resistance arose from single point mutations in either the a subunit (W206S) or the c-subunit (G20S, M23I, and A49T) of H(+)-ATPase. There is speculation of a possible association between exposure to antimalarial drugs and evolution of optochin resistance. alpha-Hemolytic streptococci resistant to optochin, particularly invasive isolates, should be tested for bile solubility or with an S. pneumoniae DNA probe before identification as viridans streptococci. PMID- 11474433 TI - Systemic effects of oral infections: a possible preventive role for the oral and maxillofacial surgeon. PMID- 11474434 TI - Complications of the mandibular sagittal split ramus osteotomy associated with the presence or absence of third molars. AB - PURPOSE: This study compared the number of unfavorable fractures after sagittal split osteotomies (SSOs) of the mandible when third molars were present or absent. PATIENTS AND METHODS: The treatment records of 262 patients (500 SSOs) were retrospectively evaluated. Group 1 consisted of 250 SSOs and concomitant removal of impacted mandibular third molars and group 2 consisted of 250 SSOs with absence of third molars. A modified SSO technique with inferior border cuts was used on all patients, and the third molars, when present, were removed after separation of the proximal and distal segments. Rigid fixation was applied using bicortical bone screws. Bone plates with monocortical screws were additionally used to secure the free bony segments of the proximal segment in cases with unfavorable fracture. RESULTS: There were 11 (2.2%) unfavorable fractures in 500 SSO procedures. The incidence of unfavorable splits was 3.2% in group 1 and 1.2 % in group 2. In group 1, all fractures occurred in teenagers, with 7 of 8 fractures extending through the extraction socket in the distal segment. Six of the 8 fractures were associated with completely impacted third molars, and 2 involved partially impacted teeth. All 3 fractures in group 2 occurred in the proximal segment. No significant difference was seen in the amount of relapse in patients with unfavorable or favorable splits. CONCLUSIONS: The occurrence of unfavorable splits is uncommon when using a modification of the SSO that includes an inferior border osteotomy. Although more unfavorable fractures occurred in teenage patients with third molars, this had no impact on the stability of the final result. PMID- 11474436 TI - Magnetic resonance imaging evaluation of temporomandibular joint disc deformities in relation to type of disc displacement. AB - PURPOSE: This study was undertaken to assess the relationship between the temporomandibular joint (TMJ) disc deformity and the type of internal derangement. PATIENTS AND METHODS: One hundred thirty-three TMJs of 72 patients (53 female and 19 male) with intracapsular dysfunction were studied using clinical and magnetic resonance imaging examinations. RESULTS: Of the 133 TMJ discs, 41.35% had no deformity, whereas 18% of the discs were folded, 19.55% were lengthened, 9.77% were round, 7.51% were biconvex, and 3.75% had thick posterior bands. The frequency of disc deformity was greater with anterior disc displacement without reduction than in cases of anterior disc displacement with reduction (P <.001). Folded and round discs were most common in cases with TMJ anterior disc displacement without reduction (P <.0001). An increase in length was seen in 56.75% of the cases with anterior disc displacement with reduction, whereas 100% of nonreducing discs were mainly folded or rounded. Crepitation was correlated with folded and round disc deformities, whereas lengthening was the feature of reducing discs associated with early and intermediate clicking. CONCLUSIONS: The results show that the degenerative changes in the TMJ disc are influenced by the degree and the type of disc displacement. The more advanced the internal derangement, the more deteriorated the disc configuration. PMID- 11474438 TI - Differences between patients with or without the need for intensive care due to severe odontogenic infections. AB - PURPOSE: Despite greatly improved dental health in industrialized countries, severe odontogenic infections still occasionally lead to hospitalization. The aim of the present study was to determine whether what symptoms, signs, or laboratory parameters on hospital admission were associated with the need for treatment in the intensive care unit (ICU). PATIENTS AND METHODS: Over an 18-month period, 100 consecutive patients (59 male, 41 female) were included in the study. Twenty percent of the patients required ICU treatment because of cardiorespiratory problems or severe complications of their infection. Both ICU and non-ICU patients were examined clinically and blood samples were taken and studied in respect to several parameters associated with infection, including C-reactive protein (CRP) levels. The findings were analyzed statistically for differences between the groups. RESULTS: No particular anamnestic background variable was associated with the need for intensive care. However, a particularly high CRP level on admission was found to be associated with a more severe course of the infection. CONCLUSIONS: This study showed that determination of CRP levels may be useful in clinical decision-making in patients with severe odontogenic infections. PMID- 11474440 TI - Prognostic factors in arthrocentesis of the temporomandibular joint: evaluation of 100 patients with internal derangement. AB - PURPOSE: The goal of this study was to analyze the prognostic factors for successful arthrocentesis for internal derangement (ID) of the temporomandibular joint (TMJ). PATIENTS AND METHODS: Arthrocentesis was carried out as the initial treatment in 103 TMJs of 100 patients with ID. The effectiveness of the treatment was evaluated in terms of the postoperative range of maximal mouth opening (MMO) and the degree of preoperative pain on a visual analog scale (VAS). Prognostic factors analyzed were age, sex, duration of painful locking, preoperative and postoperative range of MMO, preoperative degree of pain, preoperative clicking on opening mouth before becoming locked, and magnetic resonance imaging (MRI) findings. These factors were statistically analyzed. RESULTS: Seventy-three cases (71%) satisfied the criteria for success. One week after the procedure, the mean range of MMO in the successful cases (43.8 +/- 5.5 mm) was significantly greater than that in unsuccessful cases (36.9 +/- 6.6 mm) (P <.05). The mean preoperative degree of pain on a VAS in successful cases (4.7 +/- 2.9) was significantly smaller than that in unsuccessful cases (6.2 +/- 2.1) (P <.05). The preoperative incidence of morphologic changes in the mandibular condyle seen on MRI of successful cases was significantly less than that in unsuccessful cases (P <.05). CONCLUSION: Severe preoperative pain, relapse in the amount of mouth opening within 1 week after arthrocentesis, and preoperative bony changes in the condyle may be predictors of the effectiveness of arthrocentesis for ID of the TMJ. PMID- 11474442 TI - A protocol for the management of compound mandibular fractures based on the time from injury to treatment. AB - PURPOSE: The purpose of this study was to evaluate the validity of a treatment protocol for compound mandibular fractures that is based on the time of injury to treatment. PATIENTS AND METHODS: Fifty-two patients with 71 mandibular fractures were treated in a prospective fashion in conformity with the protocol. Thirty seven open reductions with rigid fixation were performed on 30 patients. The remaining 22 patients were treated solely with closed reduction and maxillomandibular fixation (MMF). Forty-five patients were treated before 72 hours and 7 after 72 hours. RESULTS: Fifty-one of the 52 patients healed without evidence of infection. One patient developed suppurative osteomyelitis. Thus, the bone infection rate was 1.9% for all patients treated and 3.3% for patients treated with rigid fixation (ORIF). CONCLUSION: These results underscore the validity of the treatment protocol to immobilize compound fractures within 72 hours of injury, if possible. If the initial treatment is delayed for more than 3 days, any infection at the compound fracture site(s) should first be resolved by MMF and intravenous antibiotics before performing an open reduction. This is done to ensure adequate perfusion of blood at the fracture site when the open reduction is performed. PMID- 11474444 TI - Modified labial tissue sliding flaps for repairing large lower lip defects. AB - PURPOSE: Common flap techniques for reconstructing large defects in the lower lip are often destructive and complex and result in unsatisfactory function and appearance. This article describes a modified technique that uses sliding labial tissue flaps to avoid these problems. PATIENTS AND METHODS: Eight patients with lower lip carcinomas had defects ranging from one third to four fifths of the length of the lower lip after tumor resection. The defects were closed with sliding labial tissue flaps, taking advantage of the elasticity of soft tissue to reduce the loss of normal tissue. Patients were followed for 2 to 12 years. RESULTS: The functional and cosmetic results were good, and there were no intra- or postoperative complications. Some patients experienced tightness of the lower lip that disappeared within 6 months. Carcinoma did not recur in any patient after a median follow-up period of more than 5 years. CONCLUSIONS: This modified labial tissue sliding flap technique is simple, safe, functionally and aesthetically satisfactory, sacrifices little healthy tissue, heals rapidly, and requires no further revision. The technique is effective for repairing defects covering one third to four fifths of the lower lip. PMID- 11474445 TI - A protocol for uvulopalatopharyngoplasty, mortised genioplasty, and maxillomandibular advancement in patients with obstructive sleep apnea: an analysis of 40 cases. AB - PURPOSE: This study evaluated the effectiveness of uvulopalatopharyngoplasty (UPPP) with mortised genioplasty and maxillomandibular advancement (MMA) for the treatment of patients with obstructive sleep apnea not controllable with appliances or continuous positive airway pressure. METHODS: Forty patients with obstructive sleep apnea were evaluated retrospectively. Thirty-three patients underwent combined UPPP and a modified mortised genioglossus advancement. Patients who had specific indications for MMA underwent combined procedures, eliminating staging of multiple surgeries. Seven patients were in this group. All patients were evaluated preoperatively and postoperatively with polysomnography to evaluate the efficacy of the treatment. RESULTS: Mean respiratory distress indices (RDI) and nadir oxyhemoglobin desaturation values were significantly improved with each of the therapies despite many patients having body mass indices significantly greater than the average quoted in other studies. Patients with moderate sleep apnea (RDI, 21 to 40) who underwent UPPP/genioglossus advancement did very well, with 86% of patients achieving success. Patients who underwent MMA all decreased their RDI by at least 56% and had an average improvement of 86%. CONCLUSIONS: The UPPP/mortised genioglossus advancement is effective for the treatment of obstructive sleep apnea. Maxillomandibular advancement is effective for treating severe sleep apnea and may, in some cases, be indicated in combination with UPPP/mortised genioglossus advancement to avoid multiple procedures. Surgical reconstruction of the upper airway is a reasonable approach to the treatment of patients with obstructive sleep apnea, and can be approached more directly to minimize repeated surgical intervention. PMID- 11474447 TI - Neural cell adhesion molecule and neurothelin expression in human ameloblastoma. AB - PURPOSE: The objective of the present study was to determine whether neural cell adhesion molecules (NCAM; CD56) and neurothelin (CD147) are expressed by adontogenic cells in the ameloblastoma. MATERIALS AND METHODS: Frozen sections from ameloblastoma specimens (n = 7) were stained with monoclonal antibody recognizing CD56 and CD147 molecules using indirect immunoperoxidase and indirect immunofluorescent techniques. RESULTS: CD56 and CD147 molecules were strongly expressed by the peripheral columnar cells of the tumor nests. Neurothelin reactivity was also present in the stellate reticulum of the nests and in some stromal components. CONCLUSIONS: The presence of these antigens in ameloblastoma supports the classic view about the neural crest origin of cells giving use to this tumor. PMID- 11474449 TI - Bone volume collected from dental implant sites during osteotomy. AB - PURPOSE: Little research has been done to determine the amount of bone harvested from implant site preparations using an inline bone collector. This study looked at the amount of bone that can be harvested from common dental implant osteotomies. PATIENTS AND METHODS: A total of 24 implants were placed in 9 patients over a 3-month period. Implant size ranged from 3.75 x 13 mm to 4.75 x 13 mm. Nine implants were placed in the maxilla, and 15 implants were placed in the mandible. Seven patients were female, and 2 patients were male. The patient age ranged from 27 to 72 years. Four patients had implants placed within 5 years after tooth extraction, and 5 patients had implants placed 5 years after tooth extraction: an analysis of variance was used to determine if there were statistical differences between maxilla versus mandible, male versus female, and edentulism less than or greater than 5 years. RESULTS: The average bone volume from the 24 osteotomies was 0.195 +/- 0.099 mL. The average osteotomy site measured 4.02 x 12.90 mm. There were no statistical differences noted among maxilla and mandible, gender, or time of edentulism. CONCLUSIONS: When using an inline bone collector to harvest implant osteotomy sites, an average of 0.195 mL of bone can be obtained from a site approximately 4.0 x 13 mm. This bone can often be combined with a xenograft or alloplastic material to provide extra bulk to fill peri-implant defects. When multiple implant sites are prepared, often sufficient bone can be obtained with the bone collector alone. PMID- 11474450 TI - Recurrent lingual swelling in an adult patient. PMID- 11474451 TI - Desired training characteristics of a potential oral and maxillofacial surgery practice associate: a New Jersey survey response. PMID- 11474454 TI - Oculo-facio-cardio-dental syndrome: two new cases. PMID- 11474455 TI - Tongue carcinoma in a young patient with dermatomyositis: a case report and review of the literature. PMID- 11474456 TI - Multiple superficial oral mucoceles: case report. PMID- 11474457 TI - A massive pleomorphic adenoma of the submandibular salivary gland accompanied by neurilemomas of the neck misdiagnosed as a malignant tumor: report of case. PMID- 11474458 TI - Third molar extractions in a patient with congenital afibrinogenemia: a case report. PMID- 11474459 TI - Solitary angiomyolipoma of the labial mucosa: a case report. PMID- 11474460 TI - Macroglossia following palatoplasty causing upper airway obstruction: case report. PMID- 11474461 TI - Schwannoma of the brachial plexus: an unusual cause for a cervical mass. PMID- 11474462 TI - An unusual penetrating facial injury due to a fishing-line sinker. PMID- 11474463 TI - Dialysis-related amyloidosis of the tongue. PMID- 11474464 TI - Ectopic thymus presenting as a submandibular swelling. PMID- 11474465 TI - Penicillium marneffei infection presenting as oral ulcerations in a patient infected with human immunodeficiency virus. PMID- 11474466 TI - Why there is a lack of female role models in oral and maxillofacial surgery. PMID- 11474467 TI - The mechanism of disc displacement. PMID- 11474468 TI - A call for uniform terminology regarding arthrocentesis. PMID- 11474469 TI - Clinical controversies in oral and maxillofacial surgery: surgical arthroscopy (part 1) or arthroscopic lysis and lavage (part 2) as the preferred treatment for internal derangement of the temporomandibular joint. PMID- 11474470 TI - Sharing the risk in the use of TMJ protheses. PMID- 11474471 TI - The renal metabolism of insulin: urinary insulin excretion in patients with mutant insulin syndrome (insulin Wakayama). AB - Many studies have shown that the kidney plays an important role in the metabolism of many proteins and small peptides. To understand insulin handling in the kidney, we examined urinary insulin excretion under several conditions in patients with mutant insulin syndrome (MIS; insulin Wakayama). Urinary excretion of insulin was studied using high-performance liquid chromatography analysis in patients with MIS. In these patients, most of the insulin extracted from a 24 hour urine collection and from urine collected after stimulation of insulin secretion by glucose or glucagon was normal insulin, whereas 90% of serum insulin is structurally abnormal (Leu-A3 insulin). On the other hand, arginine, which is known as an inhibitor of renal tubular reabsorption, increased urinary excretion of Leu-A3 insulin. The ratio of Leu-A3 and normal insulin in urine after arginine was similar to that in serum. A large amount of Leu-A3 insulin is excreted in urine when reabsorption of insulin at renal tubules is inhibited by arginine. These data indicate that normal and Leu-A3 insulin are filtered through the glomerulus with relatively little restriction. Using the fact that basal urine has a high concentration of normal insulin and an extremely low concentration of Leu-A3 insulin, which has less receptor-binding affinity, we speculated some possibilities. One possibility is that both forms of insulin are reabsorbed by the tubular cells, but with different efficiencies. Leu-A3 insulin absorption in more complete, and this suggests differences in the uptake pathways that may account for the differences in response to arginine infusions. Another possibility is that only normal insulin is secreted from tubules into urine which is mediated by receptors. Our results provide new insight into renal metabolism of insulin and showed that MIS is a useful model for studying it. PMID- 11474472 TI - The effects of treatment with alpha-lipoic acid or evening primrose oil on vascular hemostatic and lipid risk factors, blood flow, and peripheral nerve conduction in the streptozotocin-diabetic rat. AB - Oxidative stress and defective fatty acid metabolism in diabetes may lead to impaired nerve perfusion and contribute to the development of peripheral neuropathy. We studied the effects of 2-week treatments with evening primrose oil (EPO; n = 16) or the antioxidant alpha-lipoic acid (ALA; n = 16) on endoneurial blood flow, nerve conduction parameters, lipids, coagulation, and endothelial factors, in rats with streptozotocin-induced diabetes. Compared with their nondiabetic littermates, untreated diabetic rats had impaired sciatic motor and saphenous sensory nerve-conduction velocity (NCV; P <.001), reduced endoneurial blood flow (P <.001), and increased serum triglycerides (P <.01), cholesterol (P < 0.01), plasma factor VII (P <.0001), and von Willebrand factor (vWF; P <.0001). Plasma fibrinogen and serum high-density lipoprotein concentrations were not significantly different. Treatment with either ALA or EPO effectively corrected the deficits in NCV and endoneurial blood flow. ALA was associated with marked and statistically significant decreases in fibrinogen, factor VII, vWF, and triglycerides (P <.01, paired t tests before v after treatment). In contrast, EPO was associated with significant (P <.05) increases in fibrinogen, factor VII, vWF, triglycerides, and cholesterol and a significant decrease in high-density lipoprotein. Changes in levels of coagulation factors and lipids, qualitatively similar to those found with EPO, were obtained with a diet containing sunflower oil (to control for calorific and lipid content) or with a normal diet alone. Blood glucose and hematocrit levels were not significantly altered by treatments. These data suggest that although both ALA and EPO improve blood flow and nerve function, their actions on vascular factors differ. The marked effects of ALA in lowering lipid and hemostatic risk factors for cardiovascular disease indicate potential antithrombotic and antiatherosclerotic actions that could be of benefit in human diabetes and merit further study. PMID- 11474473 TI - Blunted glucose metabolism in anorexia nervosa. AB - Only few studies have specifically investigated diet-induced thermogenesis in anorexia nervosa. Twenty women, 10 anorectics (body mass index [BMI] = 14.98 +/- 1.02 kg/m(2)) and 10 controls (BMI = 22.53 +/- 0.75 kg/m(2)) were studied. Body composition was evaluated by isotopic dilution. Respiratory gas exchange was measured by indirect calorimetry. An oral glucose load (75 g) was administered to the anorectics (A) and the controls (CA). The controls underwent a second load (CB) with a higher glucose amount (1.85 +/- 0.11 g/kg body weight [BW]) to compare with the load taken by anorectics. Glucose-induced thermogenesis (GIT) was computed for 300 minutes following the load as the percent increase of energy expenditure (EE) above resting-EE (REE). Serum glucose levels were lower in anorectic patients both in fasting (3.46 +/- 0.66 v 5.23 +/- 0.23 in CA, P <.01 v 5.32 +/- 0.34 mmol in CB, P <.01) and in the postprandial state (glucose area under the curve [AUC] 175.51 +/- 6.40 v 289.80 +/- 7.30 in CA, P <.01 v 324.65 mmol in CB, P <.001); insulin AUC was lower, 1,926 +/- 452 versus 41,148 +/- 2,071 in CA, P <.0001 versus 60,765.5 pmol in CB, P <.0001. REE, normalized by fat-free mass (FFM), was similar between groups. GIT was lower in anorectics (3.58 +/- 1.20 v 5.45 +/- 1.83 in CA, P <.05 v 9.09% +/- 1.05% in CB, P <.01). Glucose oxidation was higher in anorectics than in CA (689.44 +/- 72.22 v 333.32 +/- 32.98 micromol/L/min, P <.001), but similar to CB. Lipid oxidation become negative after 30 minutes in anorectics (postprandial lipid oxidation = -93.58 +/ 39.86 v 370.61 +/- 21.73 in CA, P <.0001 v 119.01 +/- 12.32 micromol/L/300 min in CB, P <.0001). Anorectic patients displayed a low REE and GIT. Carbohydrate oxidation was similar between groups; lipid oxidation was extremely reduced. An increased protein catabolism was observed. PMID- 11474474 TI - Cardiovascular risk factors in men: The role of gonadal steroids and sex hormone binding globulin. AB - Males have higher risk of cardiovascular disease (CVD) than premenopausal females. Gonadal steroids are probably involved in the gender difference in CVD, but previous results have been conflicting. We investigated the associations between CVD risk factors and sex hormones in a cross-sectional designed study of 508 healthy males, aged 41 to 72 years. We determined total testosterone (T), sex hormone-binding globulin (SHBG), free androgen index (FAI), and estradiol (E2) and studied their relationship to body fat mass (BF), blood pressure (BP), aortic compliance, left ventricular mass (LVM), and plasma lipids (total cholesterol, high-density lipoprotein [HDL], low-density lipoprotein [LDL], very--low-density lipoprotein [VLDL], and triglycerides). In quartile analyses after adjustment for confounders (age, body mass index [BMI], alcohol consumption, and smoking), SHBG and E2 were positively associated with HDL, while FAI was negatively associated with HDL. T and SHBG were negatively associated with VLDL and triglycerides, while FAI was positively associated with VLDL and triglycerides. T and SHBG were negatively associated with BMI and BF, while FAI and E2 were positively associated with BMI and BF. E2 was negatively associated with LVM. No hormone varied with total cholesterol, LDL, BP, and aortic compliance in the adjusted analyses. In multiple regression analyses, SHBG was the main predictive variable of HDL, VLDL, and triglycerides explaining 12%, 17%, and 17% of the variation, respectively. No other hormones were selected as predictive variables for VLDL and triglycerides, but E2, T, and FAI were selected in the HDL regression, explaining 3%, 2%, and less than 1%, respectively. Our regression analyses illustrate the diverging results when investigating associations between gonadal steroids and lipids with and without SHBG adjustment. Atherogenic lipid profile in males is associated with low SHBG, low T levels, and a high FAI. Males with high E2 levels may have a less atherogenic lipid profile and lower LVM. SHBG is a key hormone in the association between sex hormones and plasma lipids. We suggest that conflicting results of cross-sectional and intervention studies of sex hormones and lipids, in part, may be explained by interindividual differences or changes in SHBG. Thus, further studies on the potential role of SHBG in the development of ischemic heart disease (IHD) should be performed. PMID- 11474475 TI - Synergy of sibutramine and low-dose leptin in treatment of diet-induced obesity in rats. AB - Tachyphylaxis to the effects of anorexigenic agents, such as sibutramine (S), may be due, in part, to counterregulatory decreases in energy expenditure (EE) and increases in hunger that result from reduced circulating leptin (L) due to loss of body fat and lowered L production/adipocyte. The present study was conducted to test the hypothesis that L administered at low doses sufficient to restore ambient L to preweight loss concentrations would enhance the intercurrent efficacy of S by reducing the strength of physiologic counterregulation to weight loss. Forty male Sprague-Dawley rats were fed a high-fat (HF) diet (45% energy) to induce obesity. After 8 weeks, the obese rats (600 +/- 58 g) were weight matched into 4 groups (N = 8/group) and implanted subcutaneously (SC) with 2 mL, 7-day Alzet mini-pumps that provided: vehicle (V, saline), L (0.5 mg/kg/d), S (3 mg/kg/d), or L+S. Food intake (FI) on the HF diet was measured daily. On day 7, 24-hour EE was measured by indirect calorimetry, and the animals then killed for body composition analysis. Compared with vehicle, treatment with S alone, but not L alone, produced significant weight loss (-23 +/- 26 v -6 +/- 16 g, P <.01). L alone, or with S, increased fat oxidation (decreased respiratory quotient [RQ]) compared with V (P <.05). The lack of decline in EE with S may be due to its documented effect to stimulate thermogenesis. Administration of L with S synergistically decreased FI and increased weight loss and fractional fat loss. A reduction in plasma L concentration may contribute to the "plateau phenomenon" observed in studies of weight loss therapies. Replacement doses of L during S administration increased weight loss and fractional fat loss by (1) decreasing food intake and (2) by increasing fat oxidation. Such drug combinations may be useful in the treatment of human obesity. PMID- 11474476 TI - The Ser(447)-Stop polymorphism of lipoprotein lipase is associated with variation in longitudinal serum high-density lipoprotein-cholesterol profiles: the Bogalusa Heart Study. AB - The Ser(447)-Stop polymorphism of lipoprotein lipase (LPL) has been associated with altered high-density lipoprotein-cholesterol (HDL-C) and triglyceride (TG) levels at individual measurements, but nothing is known of its associations with lipid profiles derived from serial measurements. We used multilevel statistical models to study effects of this polymorphism on longitudinal lipid profiles in 1,006 Bogalusa Heart Study subjects examined 4 to 9 times between the ages of 4 and 38 years. Stop(447) allele frequencies in African Americans (0.053 +/- 0.011) and whites (0.091 +/- 0.009) differed significantly (chi(2) = 7.595, 1 df, P =.006; Stop(447) homozygotes and heterozygotes combined). Overall, TG levels were lower and HDL-C levels higher in blacks than in whites of the same age and sex. Longitudinal TG profiles were lower in Stop(447) carriers at all ages. However, longitudinal HDL-C profiles differed among genotype groups with age: the Stop(447) allele was associated with higher HDL-C only in subjects above approximately 10 years of age. Genotype-specific HDL-C profiles also differed significantly among race/sex groups. Thus, we found evidence of LPL genotype effects that vary within individuals with age. Possible mechanisms, which could account for age-related changes in the effects of LPL variants, are discussed. PMID- 11474477 TI - Pycnodysostosis: clinical, radiologic, and endocrine evaluation and linear growth after growth hormone therapy. AB - Pycnodysostosis is a rare hereditary bone abnormality with an autosomal recessive mode of inheritance. We report the clinical, radiologic, and endocrine status of 8 children with this rare disease. All patients had the characteristic phenotype of the disorder including short stature (8 of 8), increased bone density (7 of 8), separated cranial sutures (8 of 8), large fontanel with delayed closure (8 of 8), obtuse mandibular angle (8 of 8), delayed teeth eruption (8 of 8), enamel hypoplasia (7 of 8), dysplastic acromial ends of the clavicles (6 of 8), frontal bossing (6 of 8), ocular proptosis (8 of 8), and dysplastic nails (8 of 8). Developmental evaluation according to the revised Denever developmental screening showed normal motor, fine motor-adaptive language, and personal social abilities in all the children. All had normal hepatic and renal functions. Serum calcium and phosphorus concentrations were normal. Two children had low serum alkaline phosphatase concentration. Short stature is a characteristic feature of pycnodysostosis. Seven of the 8 children were born short (length standard deviation score [SDS] = -3 to -1.5). Deceleration of linear growth was significant during the first 3 years of life. All the children had height SDS below -3 at the end of their third year of life. Although short stature is a feature of this genetic disorder, defective growth hormone (GH) secretion in response to provocation with clonidine and glucagon was found in 4 of the 8 patients. These 4 patients had pituitary hypoplasia on the magnetic resonance imaging (MRI) of their brain. In addition, 3 of these 4 patients had demyelination of the cerebrum. Patients with pycnodysostosis (n = 8) had low circulating concentrations of insulin-like growth factor-1 (IGF-1) compared with normal age-matched short children with constitutional short stature (CSS). IGF-I increased significantly after injecting GH for 3 days in these patients. Physiologic replacement with GH (18 U/m(2)/week) divided in daily evening doses subcutaneously increased IGF-1 concentration and improved linear growth velocity and height standard deviation scores (HtSDS) in the 4 children with GH deficiency. These data ruled out GH resistance and proved the usefulness of GH therapy in the management of short stature in these patients. In summary, some patients with pycnodysostosis have partial GH deficiency and low IGF-1 concentration. GH therapy markedly increases IGF-I secretion and improves their linear growth. MRI study of the brain including the hypothalamic-pituitary area is recommended in these children because of the high incidence of pituitary hypoplasia and cerebral demyelination. PMID- 11474478 TI - Effect of alendronate on bone mineral density in male idiopathic osteoporosis. AB - Idiopathic osteoporosis in men is an increasingly recognized disorder accounting for up to 200,000 hip fractures worldwide each year. Although there is no widely accepted or proven efficacious treatment for men with idiopathic osteoporosis, we attempted to examine the effectiveness of alendronate in this disorder. We retrospectively compared the clinical records of male patients with osteopenia (hip or spine T scores less than -1.0, with or without low-trauma fractures) treated either with alendronate 10 mg orally/day and calcium and vitamin D replacement versus conservative treatment with calcium and vitamin D alone. Review included analysis of laboratory studies and bone turnover markers in a subset of patients. We documented bone mineral density (BMD) by dual-energy x-ray absorptiometry (DXA) and repeated BMD after an average follow-up of 1.9 and 2.7 years in the alendronate-treated and conservative treatment groups, respectively. At baseline, conservatively-treated and alendronate-treated patients had similar BMD at the lumbar spine and hip. Over the period of observation, the conservatively-treated patients exhibited insignificant changes in BMD at all measured sites. In contrast, alendronate treatment resulted in a significant increase in BMD of the spine (+4.6%, P =.002), trochanter (+6.4%, P =.002), and total hip (+4.7%, P =.002). Indeed, compared with conservative treatment, alendronate-treated patients sustained a significant annualized percent increment of the BMD in the spine (2.7 +/- 0.6 v 1.1 +/- 0.3, P =.025), trochanter (4.7 +/- 1.7 v 0.7 +/- 0.6, P =.025), and total hip BMD (3.3 +/- 0.9 v 0.1 +/- 0.4, P =.0009). These data are among the first that illustrate the potential efficacy of alendronate in the management of idiopathic osteoporosis in men. PMID- 11474479 TI - Race differences in the pattern of familial aggregation for dehydroepiandrosterone sulfate and its responsiveness to training in the HERITAGE Family Study. AB - Using a familial correlation model to assess familial influences, baseline dehydroepiandrosterone sulfate (DHEAS) and its change (post-training minus baseline) in response to a 20-week endurance exercise training program were analyzed in 85 black families who participated in the HERITAGE Family Study (HERITAGE). Baseline levels were adjusted for a polynomial in age, and the training response was adjusted for a polynomial in age, as well as the baseline values, within 4 sex-by-generation groups before genetic analysis. We found that the maximal heritability for baseline DHEAS reached 66% (with no sex and generation differences) in black families, which is slightly (but not significantly) higher than the estimate (58%) reported previously in 99 white families in HERITAGE. Whereas weak, but significant, familial effects (26%) for the training response were previously reported for whites in HERITAGE, they were undetectable in the present study. Furthermore, we found heterogeneity in the pattern of familial aggregation (primarily due to different spouse and parent offspring correlations) for both the baseline and its training response between blacks and whites. In conclusion, baseline DHEAS levels in blacks were also determined by substantial familial factors (just as for whites), independent of the effects of age and sex. Genetic and nongenetic familial components influencing baseline DHEAS levels in both races may be different. PMID- 11474480 TI - Stimulation of leptin release by arachidonic acid and prostaglandin E(2) in adipose tissue from obese humans. AB - The purpose of this study was to examine the effect of arachidonic acid and its metabolites on leptin formation by explants of human adipose tissue over a 48 hour incubation in primary culture. We found that arachidonic acid or prostaglandin E(2) (PGE(2)) stimulated leptin release by explants of subcutaneous adipose tissue from obese humans. The stimulatory effect of arachidonic acid on leptin formation was blocked by NS-398, a cyclooxygenase-2 (COX-2) inhibitor. There was appreciable release of PGE(2) to the medium over 48 hours, and this was inhibited by 99% in the presence of 200 nmol/L dexamethasone or 5 micromol/L NS 398. The increase in PGE(2) release correlated with induction of COX-2 activity during the 48-hour incubation. The increase in COX-2 activity was blocked by 200nmol/L dexamethasone. The level of leptin mRNA at 48 hours was reduced by 28% if PGE(2) was added in the absence of dexamethasone, while in the presence of dexamethasone, the amount of leptin mRNA was enhanced by 156%. These data suggest that when upregulation of COX-2 is blocked by dexamethasone, exogenous PGE(2) enhances both leptin release and leptin mRNA accumulation by explants of human adipose tissue in primary culture. PMID- 11474481 TI - Insulin sensitivity and sodium excretion in normotensive offspring and hypertensive patients. AB - Insulin resistance and hyperinsulinemia have been suggested to precede and promote hypertension, possibly by impairing sodium balance. We examined insulin sensitivity and the influence of acute hyperinsulinemia on sodium excretion after acute sodium loading in hypertension-prone individuals. Insulin sensitivity and sodium excretion in response to a 1,000-mL isotonic saline bolus were examined in 24 strictly normotensive offspring of at least 1 hypertensive parent, 19 controls without a family history of hypertension, and 8 untreated, young hypertensive patients. After the saline bolus, urinary sodium excretion was measured at baseline and during a 2-hour euglycemic, hyperinsulinemic clamp, and insulin sensitivity was determined. Insulin, pressor hormones, and atrial natriuretic peptide (ANP), were measured by radioimmunoassay (RIA) or high-performance liquid chromatography (HPLC). Results are given as means +/- SEM. Offspring and controls were well matched in age (23.7 +/- 0.5; 24.6 +/- 0.5 years, respectively), blood pressure (113.0 +/- 2.9/68.5 +/- 1.9; 110.6 +/- 2.5/71.7 +/- 2.2 mm Hg, respectively), bone mass index (BMI), plasma glucose, and lipid parameters. Insulin sensitivity index did not significantly differ between offspring and controls (0.102 +/- 0.012; 0.112 +/- 0.018 micromol/min/kg/body weight [BW]/pmol, respectively), but was markedly reduced in hypertensives (0.045 +/- 0.006, P <.001). In response to sodium loading, natriuresis increased significantly (P <.05) in both offspring and controls to a similar extent, despite the presence of hyperinsulinemia, but failed to increase in hypertensives. In normotensive offspring of hypertensive patients who have not yet developed any features of the metabolic syndrome, insulin sensitivity is not impaired. Acute hyperinsulinemia impairs the ability to excrete an acute sodium load in hypertensive patients, but not in offspring of hypertensives with normal insulin sensitivity. PMID- 11474482 TI - Environmental factors modulate the effect of the APOE genetic polymorphism on plasma lipid concentrations: ecogenetic studies in a Mediterranean Spanish population. AB - To investigate APOE gene--environment interaction effects on plasma lipid concentrations, we conducted a cross-sectional study in a Mediterranean Spanish population consisting of 396 men and 513 women aged 18 to 66 years. The frequency of the epsilon 4 variant was 0.071 (95% confidence interval 0.059, 0.082), confirming the lower frequency of this allele in Southern Europe. In general, the carriers of the epsilon 2 variant had lower concentrations (P <.05) of total and low-density lipoprotein cholesterol (LDL-C), carriers of the epsilon 3 variant had intermediate concentrations, and carriers of the epsilon 4 variant had higher concentrations (P <.05) in both sexes, even after multivariate adjustment for age, body mass index, alcohol consumption, tobacco smoking, physical activity, marital status, and education. However, when the homogeneity of allelic effects according to environmental factors was tested, significant interaction terms were found. In women, an important interaction between alcohol consumption and the APOE polymorphism in determining LDL-C concentrations was found (P <.003). LDL-C concentrations in female drinkers with the epsilon 2 variant were significantly lower (P <.014) than in nondrinkers with the epsilon 2 variant. Likewise, in female drinkers with the epsilon 4 variant, LDL-C concentrations were also significantly (P <.010) lower than in nondrinkers with the epsilon 4 variant. Moreover, in female drinkers, LDL-C concentrations did not differ between carriers of the epsilon 4 and the epsilon 3 variants, and in nondrinkers, LDL-C concentrations did not differ between carriers of the epsilon 2 and the epsilon 3 variants. We also found a statistically significant interaction effect (P <.001) between the APOE polymorphism and physical activity in determining high-density lipoprotein cholesterol concentrations in men. Our results indicate that environmental factors are important modulators of the effect of the APOE polymorphism on plasma lipid concentrations. PMID- 11474483 TI - Lowering plasma free fatty acids with Acipimox mimics the antidiabetic effects of the beta 3-adrenergic agonist CL-316243 in obese Zucker diabetic fatty rats. AB - We previously reported that long-term treatment of Zucker diabetic fatty (ZDF) rats with the selective beta(3) agonist CL-316243 normalizes glycemia, decreases plasma free fatty acids (FFA) concentration, improves insulin responsiveness, and increases glucose uptake, not only in brown and white adipose tissues, but also in skeletal muscles. Because muscles do not express typical beta(3) adrenoceptors, we postulated that the muscle effect was indirect and that it was possibly mediated by an activation of the glucose-fatty acid cycle. To test this hypothesis, we investigated the effects of Acipimox, a potent inhibitor of lipolysis in adipose tissue. Similar to CL-316243, Acipimox (150 mg/kg orally) markedly decreased plasma FFA, glucose, and insulin concentrations and improved glucose tolerance while reducing the insulin response in obese (350 to 400 g) ZDF rats. Plasma FFA concentrations were significantly correlated with plasma glucose and insulin concentrations (r =.72 and.83, respectively; P <.01), indicating strong metabolic relationships between these parameters. Euglycemic hyperinsulinemic clamps combined with the 2-[(3)H]deoxyglucose method revealed that Acipimox markedly improved insulin responsiveness and significantly increased glucose uptake (Rg') in the diaphragm, the heart, and various skeletal muscles. Unlike CL-316243, Acipimox did not increase glucose use in brown or white adipose tissues. This selectivity shows that it is possible to improve diabetes in obese ZDF rats without necessarily stimulating thermogenesis in adipose tissues. Thus, decreasing plasma FFA with 2 drugs (Acipimox or CL-316243) that act via different mechanisms (acute inhibition of lipolysis or chronic stimulation of FFA oxidation) is associated with increased glucose uptake in muscles and enhanced insulin responsiveness. These observations support the hypothesis that CL-316243 may indirectly stimulate glucose uptake in muscles of type II diabetic rats by first stimulating brown adipose tissue (increasing uncoupling protein content and fatty acid oxidation) and progressively decreasing the levels of circulating FFA, resulting in activation of the glucose-fatty acid cycle or other mechanisms regulating insulin responsiveness in skeletal muscles. PMID- 11474484 TI - Decreased activities of lipoprotein lipase and hepatic triglyceride lipase in patients with gout. AB - Postheparin plasma lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) activities were measured in 30 male primary gout patients as well as in control subjects. The activities of these lipolytic enzymes were significantly decreased in the patients as compared with the controls (gout v control; LPL, 5.4 +/- 0.4 v 7.9 +/- 0.9 U; HTGL, 14.6 +/- 2.0 v 17.9 +/- 3.4 U) when matched with serum triglyceride concentration. Further, LPL activity was negatively correlated with serum- and very-low-density lipoprotein (VLDL)-triglyceride in gout patients, while that of HTGL was negatively correlated with low-density lipoprotein (LDL)-triglyceride in both gout patients and control subjects. These results suggest that decreased activities of LPL and HTGL may contribute, in part, to the increased concentrations of serum-, VLDL-, and LDL-triglyceride seen in gout patients, leading to a higher risk for coronary atherosclerotic diseases in gout. PMID- 11474485 TI - Characterization of an inhibitory effect of pioglitazone on balloon-injured vascular smooth muscle cell growth. AB - This study investigates whether pioglitazone could suppress an atherogenic process such as balloon-injured carotid intimal thickening and the proliferation of vascular smooth muscle cells (VSMC). We first examined the effect of pioglitazone to determine whether it could suppress intimal thickening induced by balloon catheterization in Sprague-Dawley rats. After 14 days postcatheterization in the left common carotid artery, the neointimal layers were completely occupied by proliferated VSMC, and the area ratio of neointima to media treated with 10 mg/kg/d of pioglitazone was significantly decreased to 57%. Next, we evaluated the effect of pioglitazone on the proliferation of rat cultured VSMC. Piogliotazone dose-dependently decreased the values of DNA synthesis, total cellular protein content, phosphorylations of extracellular signal-regulated protein kinase 1/2 and mitogen-activated protein kinase kinase 1/2, and proliferative cell nuclear antigen in VSMC. Pioglitazone also inhibited the phosphorylation of Pyk2. We conclude that pioglitazone itself could be effective for suppressing the growth of VSMC and consequent carotid intimal thickening. PMID- 11474486 TI - Peroxisome proliferator-activated receptor subtype-specific regulation of hepatic and peripheral gene expression in the Zucker diabetic fatty rat. AB - Fibrates and thiazolidinediones are used clinically to treat hypertriglyceridemia and hyperglycemia, respectively. Fibrates bind to the peroxisome proliferator activated receptor (PPAR)-alpha, and thiazolidinediones are ligands of PPAR gamma. These intracellular receptors form heterodimers with retinoid X receptor to modulate gene transcription. To elucidate the target genes regulated by these compounds, we treated Zucker diabetic fatty rats (ZDF) for 15 days with a PPAR alpha-specific compound, fenofibrate, a PPAR-gamma-specific ligand, rosiglitazone, and a PPAR-alpha/-gamma coagonist, GW2331, and measured the levels of several messenger RNAs (mRNAs) in liver by real-time polymerase chain reaction. All 3 compounds decreased serum glucose and triglyceride levels. Fenofibrate and GW2331 induced expression of acyl-coenzyme A (CoA) oxidase and enoyl-CoA hydratase and reduced apolipoprotein C-III and phosphoenolpyruvate carboxykinase mRNAs. Rosiglitazone modestly increased apolipoprotein C-III mRNA and had no effect on expression of the other 2 genes in the liver but increased the expression of glucose transporter 4 and phosphoenolpyruvate carboxykinase in adipose tissue. We identified a novel target in liver, mitogen-activated phosphokinase phosphatase 1, whose down-regulation by PPAR-alpha agonists may improve insulin sensitivity in that tissue by prolonging insulin responses. The results of these studies suggest that activation of PPAR-alpha as well as PPAR gamma in therapy for type 2 diabetes will enhance glucose and triglyceride control by combining actions in hepatic and peripheral tissues. PMID- 11474487 TI - Effects of fat mass and body fat distribution on resting metabolic rate in the elderly. AB - The aim of the present study was to investigate the relationship between resting metabolic rate (RMR) and fat-free mass, fat mass, and body fat distribution in 164 women (age 60 to 85 years; body mass index [BMI], 18.5 to 35.6 kg/m(2)) and 98 men (age 60 to 85 years; BMI, 18.3 to 36.5 kg/m(2)). After an overnight fast, RMR was assessed by indirect calorimetry and body composition by bioelectrical impedance analysis. Waist-to-hip ratio (WHR) was used to determine fat distribution. Results from linear regression analysis showed that most of the variance in RMR could be attributed to fat-free mass in women (R(2) = 0.54) and men (R(2) = 0.44), respectively. Fat mass explained an additional 3% and 2% of the variability in RMR in women and men, respectively. In stepwise multiple regression analysis, considering body composition and fat distribution, only fat free mass and WHR were significant predictors of RMR in both sexes. In addition to fat-free mass, in women 6% and in men 8% of the variability in RMR was attributable to WHR. Grouping subjects according to their WHR, RMR, and RMR adjusted for fat-free mass and fat mass showed a significant increase with increasing WHR in both sexes. Results indicate that RMR not only depends on fat free mass but also is influenced by fat mass, especially by fat distribution. These findings support our hypothesis of an elevated RMR with increasing abdominal body fat as a direct consequence of its greater metabolic activity. PMID- 11474488 TI - Abdominal fat distribution in pre- and postmenopausal women: The impact of physical activity, age, and menopausal status. AB - Age-related increases in total body fat have been reported, but the impact of menopause on abdominal fat distribution is still unclear. The purpose of this study was to determine the impact of menopausal status on abdominal fat distribution using magnetic resonance imaging (MRI). In addition, we investigated the influence of abdominal fat distribution on blood lipid profiles and leptin concentrations. Twenty-three premenopausal (PRE), 27 postmenopausal (POST), and 28 postmenopausal women on estrogen replacement therapy (ERT) had measurements of regional abdominal fat, blood lipids, and serum leptin concentrations. The women were matched for body mass index (BMI) and total body fat mass. Age and menopausal status were not found to be significant predictors of total abdominal fat, visceral fat, or subcutaneous fat, while physical activity was a significant predictor (P <.01) for total abdominal fat (R(2) =.16), visceral fat (R(2) =.32) and percent visceral fat (R(2) =.25). There was a trend for a greater visceral fat content in the POST women compared with the PRE women (2,495.0 +/- 228.4 v 1,770.4 +/- 240.8 cm(2), respectively, P =.06). The percent visceral abdominal fat was significantly lower (P <.05) in the premenopausal women than in either postmenopausal group (PRE, 23.2% +/- 1.7%; POST, 28.9% +/- 1.8%; ERT, 28.9% +/- 1.6%). Menopausal status and age did not influence any of the blood lipid values. Abdominal fat distribution was a significant predictor of cholesterol concentrations and the cholesterol/high-density lipoprotein-cholesterol (HDL-C) ratio, but only accounted for approximately 15% of the variability in these levels. Total body fat and physical activity accounted for 47% of the variability in leptin concentrations, while abdominal fat distribution, age, and menopausal status were not significant predictors. In conclusion, in early postmenopausal women, the level of physical activity accounts for the variability in abdominal fat distribution observed, while menopausal status and age do not play a significant role. ERT was not associated with additional benefits in abdominal fat distribution compared with postmenopausal women not on ERT or in the blood lipid profile in these women. PMID- 11474489 TI - Effect of atorvastatin on low-density lipoprotein subtypes in patients with different forms of hyperlipoproteinemia and control subjects. AB - Atorvastatin is a potent hydroxy-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor that decreases low-density lipoprotein (LDL) cholesterol and triglyceride concentrations, but little is known about its effects on LDL subtype distribution in different types of hyperlipoproteinemia. Thus, we evaluated the influence of atorvastatin (10 mg/d, 4 weeks) on lipid concentrations and LDL subtype distribution in patients with hypercholesterolemia (n = 9; LDL cholesterol, 227 +/- 30 mg/dL; triglycerides, 137 +/- 56 mg/dL), patients with type 2 diabetes and dyslipoproteinemia (n = 11; LDL cholesterol, 163 +/- 34 mg/dL; triglycerides, 260 +/- 147 mg/dL), and controls (n = 10; LDL cholesterol, 116 +/- 20 mg/dL; triglycerides, 130 +/- 47 mg/dL). Cholesterol concentration was determined in 7 LDL subfractions isolated by density gradient ultracentrifugation before and during atorvastatin treatment. Atorvastatin decreased LDL cholesterol (-36%, -28%, and -41%, all P <.01) and triglyceride (-4%, NS; -2%, NS; -24%, P <.05) concentrations but had little effect on high-density lipoprotein (HDL) cholesterol (-1%, NS; +10%, P <.05; +6%, NS) in hypercholesterolemic, diabetic, and control subjects, respectively. In all 3 groups, a significant reduction in cholesterol in each LDL subfraction was observed. Large-buoyant (LDL-1, LDL-2) and intermediate-dense (LDL-3, LDL-4) LDL were reduced more than small-dense (LDL 5 through LDL-7) LDL in hypercholesterolemic (-45%, -35%, and -32%, P <.05) and control subjects (-48%, -44%, and -25%, P <.05), but in diabetic patients cholesterol reduction was uniform in all LDL subtypes (-32%, -27%, and -29%, P =.45). Thus, atorvastatin decreases cholesterol concentration in all LDL subfractions in hypercholesterolemic, diabetic, and control subjects. However, the relative reduction of individual LDL subtypes differed between these groups. This finding suggests that the effect of atorvastatin on LDL subtype distribution depends on the type of underlying hyperlipoproteinemia. PMID- 11474490 TI - Introduction: Modern management of acute myeloid leukemia. PMID- 11474491 TI - Current status and perspectives of therapy for acute myeloid leukemia. AB - Intensification of induction and postremission chemotherapies has resulted in a steady improvement in outcomes for adults with acute myeloid leukemia (AML) over the past two decades. The global response to initial therapy is now in the range of 60% to 80%, with 20% to 30% of patients experiencing long-term disease-free survival. Modern methodologies have revealed that cytogenetic aberrations are closely related to treatment outcome and have allowed subgroups of patients to be identified who have AML with a good, intermediate, or poor prognosis. The actions and interactions of cytogenetics and therapy in determining treatment outcome have been investigated in a series of clinical trials conducted by the German AML Cooperative Group. Evidence is presented that therapy and cytogenetics can independently influence outcome. It is hoped that our increasing understanding of the biological mechanisms that dictate disease characteristics will lead to improved outcomes for patients with AML. PMID- 11474492 TI - Managing therapy in older adult patients with acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is predominantly a disease of older adults, with more than 50% of cases occurring in adults over 60 years of age. Treatment of AML in older adults is complicated not only by comorbidities that are common in this patient population, but also by the prevalence, in this age group, of forms of AML with a poor prognosis. The problems encountered and the strategies that have been used to improve the outlook in older adults with AML are presented. The two main strategies for improving outcomes in older adults with AML are to develop effective chemotherapeutic regimens with improved tolerability, and to reduce drug resistance. In studies to identify optimal chemotherapeutic regimens in older adults, a satisfactory balance between efficacy and toxicity has not yet been achieved. Also, the use of growth factors to promote hematopoietic recovery has yet to yield consistent reductions in treatment-related morbidity or mortality. Drug resistance can be modified by inhibiting drug efflux mechanisms or by increasing sensitivity to cytotoxic agents, but these strategies have not yet been shown to significantly affect outcomes. Novel approaches including antibody-targeted and molecular-targeted chemotherapy may have the potential to improve the prognosis for older adults with AML. PMID- 11474493 TI - Postremission therapy in adults with acute myeloid leukemia. AB - A patient with acute myeloid leukemia (AML) who has achieved remission after induction chemotherapy still harbors 10(9) to 10(10), albeit undetectable, leukemic cells. Optimally, postremission therapy safely reduces the leukemic burden to a level compatible with long-term disease-free survival. Although older adults fare poorly with intensive postremission therapy, young and middle-aged adults can receive either chemotherapy based on high-dose cytarabine or myeloablative therapy in preparation for autologous or, if a histocompatible donor is available, allogeneic stem cell rescue. The rationale for each approach, the prospective trials comparing the various options, and a suggested strategy for choosing among them is presented. Although cytogenetic category at diagnosis is the most important prognostic factor, this feature remains an imperfect guide to postremission therapy. An informal consensus has arisen in favor of chemotherapy for patients with good prognosis and allogeneic transplant for those whose AML displays an adverse karyotype. For the intermediate group, an individualized decision is required and any of the three options is reasonable. Because the relapse rate is so high, new therapies for AML, such as signaling and immunotherapeutic approaches, are the focus of active investigation. PMID- 11474494 TI - Current use and future development of gemtuzumab ozogamicin. AB - The antigen CD33 is expressed on blast cells in 80% to 90% of acute myeloid leukemia (AML) cases but, importantly, is not expressed on pluripotent hematopoietic stem cells or on nonhematologic cells. Gemtuzumab ozogamicin (CMA 676) uses a recombinant humanized anti-CD33 monoclonal IgG4 antibody to deliver the potent cytotoxin, calicheamicin, into cells. Three multicenter trials have evaluated the efficacy and safety of gemtuzumab ozogamicin as a single agent in 142 patients with CD33+ AML in untreated first relapse. The median age was 61 years (range, 22 to 84 years), none had prior myelodysplasia, and all had had a first complete remission lasting > or = 3 months. Two doses of 9 mg/m2 were given 14 days apart by 2-hour intravenous infusion. The overall response rate was 30% (ie, < or = 5% blasts remaining in the bone marrow, neutrophils > or = 1,500/microL, and red blood cell and platelet transfusion independence). There was no significant difference in response rate between patients less than 60 years of age and those > or = 60 years old (34% v 26%, respectively) or between patients whose first remission had lasted less than 12 months or > or = 12 months (28% v 32%, respectively). Overall survival was 31% at 1 year; median survival was 5.9 months. Median relapse-free survival was 6.8 months. An infusion-related syndrome (chills, fever, rigors, nausea, hypotension, and pain) was common. Severe myelosuppression occurred in all patients, but severe mucositis (4%) and infections (23%) were relatively infrequent. Severe hyperbilirubinemia (23%) and elevated hepatic transaminases (18%) were usually transient. Among all 142 patients, the median total hospitalization was 24 days; 16% of patients required < or = 7 days in hospital. Additional studies are currently evaluating gemtuzumab ozogamicin in combination with, or as an alternative to, other standard AML chemotherapy. PMID- 11474495 TI - Ionizing radiation as a response-enhancing agent for CD95-mediated apoptosis. AB - CD95 (Fas/APO-1) is a death receptor on the surface of a wide variety of cell types. In most cells examined, ionizing radiation acts as a response-enhancing agent for CD95-mediated cell death. Although DNA-damaging radiation appears to modulate CD95-mediated signals through multiple mechanisms, the only well characterized mechanism is activation of the tumor-suppressor protein p53, which transcriptionally regulates the expression of CD95 on various cell types. The ligand for CD95 is expressed by activated lymphocytes and natural-killer cells, which produce factors that sensitize cells resistant to CD95-mediated cell death. Ligation of CD95 on irradiated tumor cells might be achievable using emerging modalities that reactivate the stalled anti-tumor immune response. PMID- 11474496 TI - Manganese superoxide dismutase-plasmid/liposome (MnSOD-PL) administration protects mice from esophagitis associated with fractionated radiation. AB - Intraesophageal administration of manganese superoxide dismutase-plasmid/liposome (MnSOD-PL) prior to single fraction radiation has been shown to protect mice from lethal esophagitis. In our study, C3H/HeNsd mice received fractionated radiation in two protocols: (i) 18 Gy daily for four days with MnSOD-PL administration 24 hr prior to the first and third fraction, or (ii) 12 Gy daily for six days with MnSOD-PL 24 hr prior to the first, third, and fifth fraction. Control radiated mice received either no liposomes only or LacZ (bacterial beta-galactosidase gene)-plasmid/liposome (LacZ-PL) by the same schedules. We measured thiol depletion and lipid peroxidation (LP) in whole esophagus and tested the effectiveness of a new plasmid, hemagglutinin (HA) epitope-tagged MnSOD (HA MnSOD). In fractionation protocols, mice receiving MnSOD-PL, but not LacZ-PL (200 microl of plasmid/liposomes containing 200 microg of plasmid DNA), showed a significant reduction in morbidity, decreased weight loss, and improved survival. Four and seven days after 37 Gy single fraction radiation, the esophagus demonstrated a significant increase in peroxidized lipids and reduction in overall antioxidant levels, reduced thiols, and decreased glutathione (GSH). These reductions were modulated by MnSOD-PL administration. The HA-MnSOD plasmid product was detected in the basal layers of the esophageal epithelium 24 hr after administration and provided significant radiation protection compared to glutathione peroxidase-plasmid/liposome (GPX-PL), or liposomes containing MnSOD protein, vitamin E, co-enzyme Q10, or 21-aminosteroid. Thus, MnSOD-PL administration significantly improved tolerance to fractionated radiation and modulated radiation effects on levels of GSH and lipid peroxidation (LP). These studies provide further support for translation of MnSOD-PL treatment into human esophageal radiation protection. PMID- 11474497 TI - Experimental determination of depth-scaling factors and central axis depth dose for clinical electron beams. AB - Depth-scaling factors rho(eff) for clear polystyrene and polymethylmethacrylate (PMMA) phantoms have been determined experimentally as a function of nominal electron-beam energy in the range 6 to 22 MeV. Values of rho(eff) have been calculated from the ratio rho(eff) = R(wat)(50) / R(med)(50), where R(wat)(50) and R(med)(50) are the measured depths of 50% ionization in electron solid water and plastic (clear polystyrene and PMMA) phantoms, respectively. Measurements were made using an Attix chamber in an electron solid water phantom, a Holt chamber in a clear polystyrene phantom, and a Markus chamber in a PMMA phantom. The average value of measured rho(poly)(eff) was found to be 0.999 +/- 0.009. This is higher than the value of 0.975 recommended by Task Group 25 (TG-25) of the American Association of Physicists in Medicine (AAPM) by 2.5%. Depending on energy, the maximum differences between the AAPM TG-25-recommended and the measured values lie in the range 1% to 3.5%. Similarly, the average value of measured rho(PMMA)(eff) was found to be 1.168 +/- 0.023. This is higher than the AAPM TG-25-recommended value of 1.115, by 5%. Depending on energy, the maximum differences between the AAPM TG-25-recommended and the measured values lie in the range 3% to 8%. Central axis depth dose curves in water were generated for 6, 15, and 20 MeV electron beams from measured depth-ionization data in PMMA and clear polystyrene phantoms following the recommendations of the AAPM TG-25 report and using both TG-25-recommended and experimentally determined values of depth scaling factors rho(eff). For both phantoms, either the TG-25-recommended value or the experimentally determined values of rho(eff) yielded agreement to within about 2 mm among all depth doses in water at the depths of clinical relevance. PMID- 11474498 TI - Two metachronous tumors in the radiotherapy fields of a patient with Li-Fraumeni syndrome. AB - A woman with a family history of brain tumors in her daughter and sister presented with a breast cancer. She subsequently developed two metachronous primary tumors: a small-cell lung cancer and a colon carcinoma. These tumors arose within the internal mammary radiotherapy field and within the field irradiated for ovariolysis. The p53 gene was analyzed in whole blood lymphocytes using a functional assay developed in yeast Saccharomyces cerevisiae, which tests the transcriptional competence of p53. DNA from the colon cancer cells was analyzed by polymerase chain reaction and sequencing. The patient had a germline inactivating p53 mutation, confirming the diagnosis of Li-Fraumeni syndrome (LFS). The colon tumor and the lung tumor both conserved the mutant p53 allele but had lost the wild-type allele. This observation and the experimental data suggest an abnormal sensitivity of LFS patients to radiogenic carcinogenesis. The indications and extent of radiotherapy in patients with a clinical or molecular diagnosis of LFS should be discussed individually and should take into account the risk of secondary neoplasms arising in the radiation fields. PMID- 11474499 TI - Unresectable hepatocellular carcinoma treated with radiotherapy and/or chemoembolization. AB - The purpose of our study was to evaluate the outcome, patterns of failure, and toxicity for patients with unresectable hepatocellular carcinoma (HCC) treated with radiotherapy, transcatheter arterial chemoembolization (TACE), or combined TACE and radiotherapy. Forty-two patients with unresectable HCC were treated with combined radiotherapy and TACE (TACE+RT group, 17 patients), radiotherapy alone (RT group, 9 patients), or with TACE alone (TACE group, 16 patients). Mean dose of radiation was 46.9 +/- 5.8 Gy in a daily fraction of 1.8 to 2 Gy, directed only to the cancer-involved areas of the liver. TACE was performed with a combination of Lipiodol, doxorubicin, cisplatin, and mitomycin C, followed by Gelfoam or Ivalon embolization. Tumor size was smaller in the TACE group (mean: 5.4 cm) compared with the TACE+RT group (8.6 cm) and the RT group (13.1 cm) (P = 0.0003). The median follow-up was 24 months in the TACE+RT group, 28 months in the RT group, and 23 months in the TACE group. Survival was significantly worse for patients treated with radiotherapy alone due to the selection bias of patients with more advanced disease and compromised condition in this group. In contrast, the TACE+RT and TACE groups had comparable survival (two-year rates: TACE+RT 58%, TACE 56%, P = 0.69). The local control rate for the treated tumors was similar in the TACE+RT and TACE groups (P = 0.11). The intrahepatic recurrence outside the treated tumors was common and similar between these two groups (P = 0.48). The extrahepatic progression-free survival was significantly shorter for patients in the TACE+RT group than in the TACE group (two-year rates: TACE+RT 36%, TACE 100%, P = 0.002). Seven patients died from complications of treatment. Local radiotherapy may be added to treat patients with unresectable HCC, and the control of progression of the treated tumors was promising even in patients with large hepatic tumors. Survival of patients with combined TACE and radiotherapy was similar to that with TACE as the only treatment, while a significant portion of the patients treated with radiotherapy developed extrahepatic metastasis. PMID- 11474500 TI - Long-term results of neoadjuvant radiation therapy for breast cancer. AB - Our aim was to determine the long-term outcome and the possible role of neoadjuvant (preoperative) radiation therapy for breast cancers unsuitable for primary conservative surgery. From 1977 to 1992, 75 unifocal non-inflammatory and non-metastatic T2 and T3 breast cancers were treated in our department. All these patients underwent initial radiotherapy, followed by secondary limited surgery. A population of 74 patients, aged from 32 to 82 years (median 56 years), presenting 49 T2 and 26 T3 tumors, was studied. Seventy-two patients (96%) underwent secondary tumorectomy and three patients (4%) reduction mammaplasty. The secondary tumorectomy was followed by a postoperative boost. There were nine recurrences, treated by mastectomy in eight cases and by tumorectomy in one case. Twenty-five patients showed secondary dissemination. Forty-seven patients are still alive and free of disease. The cosmetic results were considered excellent or satisfactory in 71 cases. Under good conditions, preoperative radiotherapy (as well as preoperative chemotherapy) allows the possibility of conservative surgery for cancers of more than 3 cm. The choice between the two modalities depends on the patient's condition and on a precise analysis of all prognostic factors that would justify the need for systemic treatment. PMID- 11474501 TI - Plant cutin biosynthesis: the involvement of a new acyltransferase. PMID- 11474503 TI - Re: Buduhan G. McRitchie DI. Missed injuries in patients with multiple trauma. J Trauma. 2000;49:600--605. PMID- 11474502 TI - Infection and cancer. PMID- 11474505 TI - [Cutaneous and mucosal infections caused by Candida albicans. Epidemiology, diagnosis, treatment]. PMID- 11474504 TI - Re: Lanoix R et al. C-spine injury associated with gunshot wounds to the head: retrospective study and literature review. J Trauma.2000;49:860--863. PMID- 11474506 TI - [Cutaneous ectoparasitic infections. Epidemiology, diagnosis, treatment]. PMID- 11474507 TI - [Dermatophyte infection of the smooth skin and folds. Diagnosis and treatment]. PMID- 11474508 TI - [Diagnostic approach in pruritus. Excluding anal and vulval pruritus]. PMID- 11474509 TI - [Cutaneous and mucosal herpes. Epidemiology, physiopathology, diagnosis, clinical course, treatment]. PMID- 11474511 TI - [Primary and secondary syphilis. Epidemiology, diagnosis, treatment]. PMID- 11474510 TI - [Varicella and zona. Epidemiology, physiopathology, diagnosis, clinical course, treatment]. PMID- 11474512 TI - [Gonococcal and Chlamydia trachomatis urogenital infections (excluding Nicolas Favre disease). Epidemiology, diagnosis, clinical course, treatment]. PMID- 11474513 TI - [Dermatologic signs of sarcoidosis]. PMID- 11474514 TI - [Sore]. PMID- 11474515 TI - [Cutaneo-mucosal manifestations of HIV primary infection]. PMID- 11474516 TI - [Soft tissue infection caused by anaerobic germs. Etiology, diagnosis, treatment]. PMID- 11474517 TI - [Cutaneo-mucosal manifestations of systemic lupus erythematosus]. PMID- 11474518 TI - [Systemic lupus erythematosus. Diagnosis, clinical course, treatment principles]. PMID- 11474519 TI - [Normal and pathological cicatrization. Physiology and anatomopathology]. PMID- 11474520 TI - [Dermatologic topical corticosteroids. Principles and usage standards, posology]. PMID- 11474521 TI - [Acne. Diagnosis, physiopathology and treatment]. PMID- 11474522 TI - [Diagnostic approach in bullous dermatosis in the adult]. PMID- 11474523 TI - [Human papilloma virus (HPV) infections of the skin and mucous membranes. Epidemiology, physiopathology, diagnosis, treatment]. PMID- 11474525 TI - [Diagnostic approach in ulceration or erosion of oral and genital mucous membranes]. PMID- 11474526 TI - [Purpura. Diagnostic approach]. PMID- 11474527 TI - [Diagnostic approach in erythema]. PMID- 11474529 TI - [Contact eczema. Physiopathology, diagnosis, etiology, clinical course, treatment]. PMID- 11474528 TI - [Drug-induced dermatoses. Diagnosis]. PMID- 11474530 TI - [Atopic dermatitis. Diagnosis, clinical course, treatment]. PMID- 11474531 TI - [Urticaria and Quincke's edema. Diagnosis, etiology, treatment]. PMID- 11474532 TI - [Leg ulcer]. PMID- 11474533 TI - [Diagnostic approach in erythema nodosum]. PMID- 11474534 TI - [Epithelial skin cancer. Epidemiology, etiology, pathologic anatomy, diagnosis, treatment principles]. PMID- 11474535 TI - [Melanoma. Epidemiology, diagnosis, clinical and histopathologic criteria of prognosis, clinical course and screening]. PMID- 11474536 TI - [Psoriasis. Diagnosis, clinical course, treatment principles]. PMID- 11474537 TI - [Bacterial skin infections: impetigo, furunculosis, erysipelas. Etiology, diagnosis, clinical course, treatment]. PMID- 11474538 TI - Abstracts of the 9th International Magnesium Symposium. Vichy, France, September 10-15, 2000. PMID- 11474539 TI - Stimulatory effect of D-ephedrine on beta3-adrenoceptors in adipose tissue of rats. AB - The effect of ephedrine on beta3-adrenoceptos (beta3-AR) was studied in the isolated adipose tissue of Wistar rat. Incubation with D-ephedrine (0.1-10 microM) induced a concentration-dependent decrease of uptake of [14C]-deoxy-D glucose into white adipose tissues (WAT). The inhibitory effect of D-ephedrine was potentiated by BRL 37344, the agonist of beta3-AR and concentration dependently inhibited by SR 59230A, the selective antagonist of beta3-AR. The action of D-ephedrine on beta3-AR was further blocked by the antibodies for beta3 AR, but not the immunoglobulin. in a concentration-dependent manner. Moreover, D ephedrine increased glycerol release from the isolated brown adipose tissues (BAT) and this action was also abolished by SR 59230A at concentration sufficient to block beta3-AR. Thus, these results suggest that D-ephedrine has the ability to activate beta3-AR both in WAT and BAT of Wistar rats in vitro. PMID- 11474540 TI - Vagal neurostimulation in patients with coronary artery disease. AB - We tested the hypotheses that (1) progression of coronary artery disease (CAD) increases sympathetic inflow to the heart, thus impairing cardiac blood supply, and (2) reduced sympathetic tone improves cardiac microcirculation and ameliorates severity of anginal symptoms. Electrical irritation of the nerve auricularis--a sensitive ramus of the vagus nerve--provides a central sympatholytic action. Using this technique, we studied the effects of vagal neurostimulation (VNS) on hemodynamics, the content of atrial noradrenergic nerves and the microcirculatory bed of CAD patients. VNS was performed in the preoperative period of CAD patients with severe angina pectoris. The comparison groups consisted of untreated patients with CAD or Wolff-Parkinson-White syndrome. Atrial tissue of patients with this syndrome (n = 6); with effort angina (n = 14); with angina at rest (n = 10); and with severe angina treated with VNS (n = 8) contained the following volume percentages of noradrenergic nerves: 1.7+/-0.1%, 1.3+/-0.3%, 0.5+/-0.1% (p < 0.05 vs. the other groups) and 1.3+/-0.2%, respectively. In these groups, cardiac microcirculatory vessels (diameter, 10-20 microm) had the following densities: 2.7+/-0.2%, 3.4+/-0.2%, 2.0+/-0.4% (p < 0.05 vs. the other groups) and 3.3+/-0.3%, respectively. VNS treatment abolished angina at rest, decreased heart rate and blood pressure. It improved left ventricular ejection fraction from 50+/-1.5% to 58+/-1.0% (p < 0.05), also changing left ventricular diastolic filling. The ratio of time velocity integrals of the early (Ei) to late (Ai) waves increased from 1.07+/ 0.12 to 1.65+/-0.17 after VNS (p < 0.05). In electrocardiograms of VNS-treated patients, QRS- and QT-duration were shortened. the PQ-interval did not change, but T-wave configuration improved. In the postoperative period, heart failure occurred in 90% of the control group. vs. 12% in patients treated with VNS (p < 0.05). We conclude that CAD is characterized by overactivity of sympathetic cardiac tone. Vagal stimulation reduced sympathetic inflow to the heart, seemingly via an inhibition of norepinephrine release from sympathetic nerves. VNS' sympatholytic/vagotonic action dilated cardiac microcirculatory vessels and improved left ventricular contractility in patients with severe CAD. PMID- 11474541 TI - Irregular activation of individual sweat glands in human sole observed by a videomicroscopy. AB - Sweat secretion from individual sweat glands on the human sole was observed in four male subjects by using a videomicroscope and correlated with sudomotor neural activity recorded from the tibial nerve by means of microneurography. Individual sweat glands could be distinguished as active, less active and inactive according to the incidence of sweat secretion during spontaneous sweating. The threshold amplitude of the sudomotor burst necessary for sweat secretion varied from gland to gland. The number of sweat secretion was significantly related to the threshold amplitude. Sweat glands often failed to produce sweat secretion even when a suprathreshold burst occurred: only 46.1+/ 3.8% (mean +/- S.E.M.) of the suprathreshold bursts elicited sweat secretion. Failure of the sweat secretion tended to appear after several bursts occurred consecutively with short intervals. In spite of the variability in sweat gland activity, the number of sweat glands recruited was linearly related to the amplitude of the sudomotor burst (P < 0.001). Thus, although sweat secretion from each sweat gland depends primarily on the intensity of sudomotor neural activity. the activity of each sweat gland may fluctuate temporally as the result of irregular activation of sudomotor fibers and possibly some intrinsic factors of the gland. PMID- 11474542 TI - Depression of transmitter release at synapses in the rat superior cervical ganglion: the role of transmitter depletion. AB - The characteristics of depression of the excitatory postsynaptic potential (EPSP) during a short train of impulses to the rat superior cervical ganglion (SCG) have been ascertained with the object of determining the relative contributions of transmitter depletion and autoreceptors to depression. Successive EPSPs in a short train were depressed after the first (Vo) up to about the fourth impulse when a steady-state depressed EPSP level (Vss) was reached. Vss increased with the stimulation frequency between 1 and 30 Hz. Vo recovered after a short train with a time constant of about 2.8 s in the frequency range from 5 to 30 Hz. In order to determine if depression was related to changes in calcium influx with successive impulses in the train. preganglionic boutons were loaded with the calcium indicator Oregon Green 488 BAPTA-1 and line scans taken through individual boutons with a confocal laser microscope. Successive calcium transients were of about the same amplitude in boutons during short trains of impulses at 5 Hz. The contribution of autoreceptors activated by the action of endogenously derived adenosine on the extent of depression of the EPSP during short trains was ascertained by blocking these receptors with 8 phenyltheophylline (10 microM). There was no change in the extent or time course of development of depression. Similar results were obtained with the opioid receptor antagonist naloxone (10 microM) and the adrenergic receptor antagonist yohimbine (10 microM). Factors, which increased the extent of transmitter release during a train, such as increasing the external calcium concentration from 0.8 to 2.5 mM, increased depression. Factors. which decreased the extent of transmitter release such as increasing the exogenous adenosine concentration between 1 and 200 microM decreased depression. These results are interpreted in terms of a model in which vesicles are mobilised by a calcium-dependent process from a store into an available pool of docked vesicles. Depletion of the docked vesicles during exocytosis then leads to depression of transmitter release during a train of impulses. PMID- 11474543 TI - Estrogen blocks the cardiovascular and autonomic changes following vagal stimulation in ovariectomized rats. AB - The current investigation examines the effect of acute and chronic estrogen administration on baroreflex sensitivity and autonomic tone following 2 h of vagal afferent stimulation in ovariectomized female rats. Female Sprague-Dawley rats were ovariectomized and supplemented daily for 7 days with either estrogen (OVX-E2; 0.5 microg/kg; s.c.) or saline (OVX-S; 0.9%; s.c.). On the 8th day the animals were anaesthetized (sodium thiobutabarbitol; 100 mg/kg) and instrumented for recording blood pressure, heart rate and efferent vagal and renal nerve activities. The baroreflex was evoked using intravenous injection of various doses of phenylephrine hydrochloride (0.025, 0.05+/-0.1 mg/kg). Electrical stimulation of vagal afferents for 2 h produces autonomic imbalance characterized by sympathoexcitation and parasympathetic withdrawal. This protocol of vagal stimulation produced a significant increase in renal nerve activity (from 20+/-6 to 140+/-20 spikes/2 s) and decreases in both vagal nerve activity (from 22+/-3 to 10+/-2 spikes/2 s) and baroreflex sensitivity (from 0.55+/-0.05 to 0.3+/-0.05) in OVX-S female rats. However. vagal stimulation had no effect on baroreflex sensitivity or autonomic nerve activities in OVX-E2 rats. Administration of a single, bolus dose of estrogen (1 x 10(-2) mg/kg) to OVX-S rats immediately prior to termination of vagal stimulation blocked the changes in autonomic nerve activities and baroreflex sensitivity previously observed. These results suggest that both chronic and acute estrogen supplementation may provide resistance to the autonomic disturbances associated with visceral afferent activation. PMID- 11474544 TI - Low pH modulation of recombinant vanilloid receptors and perivascular capsaicin sensitive sensory neurotransmission. AB - The effect of low pH on capsaicin-sensitive sensory neurotransmission in the rat isolated mesenteric arterial bed and at recombinant (rVR1) vanilloid receptors was investigated. Mesenteric sensory neurogenic vasorelaxation elicited by electrical field stimulation was reversibly inhibited by lowering pH from 7.4 to 6.9 and 6.3. Capsaicin-induced vasorelaxation was not different at pH 6.9, but was attenuated at pH 6.3. Vasorelaxation to calcitonin gene-related peptide, the principal sensory motor neurotransmitter in rat mesenteric arteries, was not different at pH 6.9 or pH 6.3. In rVR1-transfected HEK293 cells, acidic conditions enhanced the affinities of capsaicin and capsazepine at rVR1, but did not affect the potency of carbachol at endogenous muscarinic receptors. Following inactivation of endogenous acid-sensitive ion channels, lowering pH (6.0-4.5) directly increased [Ca2+]i in rVR1-HEK293 cells (EC50 5.5). This response was abolished by 1 microM capsazepine. In conclusion, a decrease in pH (to 6.9 and 6.3) enhances the affinity of capsaicin at rVR1, but inhibits sensory neurotransmission in the rat mesenteric arterial bed. This likely explains why there is no evidence of an enhancement of sensitivity to capsaicin at endogenous vanilloid receptors, as observed with rVR1. When pH is reduced still further (6.0 5.5) there is direct activation of rVR1. PMID- 11474545 TI - Sympathetic efferent pathways projecting to the vagina in the dog. AB - The level of the thoracolumbar sympathetic outflow projecting to the vagina and the laterality of the signals passing through the splanchnic nerves to the vagina have not been well understood. This study was undertaken to explore the sympathetic innervation of the canine vagina by measuring intraluminal pressure of the vagina and contraction of the vaginal wall by electrical stimulation of each of the thoracolumbar splanchnic nerves with or without transection of unilateral hypogastric nerve (HGN). The 2nd-4th lumbar splanchnic nerve (LSN) stimulation elicited elevation of intraluminal pressure of the vagina and contraction of the vaginal wall. In contrast, intermesenteric plexus stimulations caused no response in both the vaginal pressure and the wall itself. On the right side, 12 of 27 LSNs that were stimulated elicited elevation of vaginal pressure and 16 of 32 LSN stimulations caused contraction of the wall, 10 increases in tension and 6 reductions. On the left side, 14 of 26 LSN stimulations elicited elevation of vaginal pressure and 12 of 27 LSN stimulations caused contraction of the wall, 5 increases in tension and 7 reductions. After transection of the right HGN, 6 of 14 right LSN stimulations elicited elevation of vaginal pressure and 10 of 15 right LSN stimulations caused contraction of the wall, 9 increases in tension and 1 reduction. After transection of the left HGN, 13 of 19 left LSN stimulations elicited elevation of vaginal pressure and 10 of 18 left LSN stimulations caused contraction of the wall. 4 increases in tension and 6 reductions. The results indicate that each of the L2-L4 LSNs on either side sends signals to the canine vagina via multiple routes with a crossing site at the level of the caudal mesenteric plexus (CMP). PMID- 11474546 TI - Nociceptive stimulation increases NO synthase mRNA and vasopressin heteronuclearRNA in the rat paraventricular nucleus. AB - Nociceptive stimulation causes neuroendocrine responses such as arginine vasopressin (AVP) release and activation of the hypothalamo-pituitary-adrenal (HPA) axis. We examined the effects of nociceptive stimulation on the expression levels of neuronal nitric oxide synthase (nNOS) mRNA, heteronuclear (hn)RNA for AVP and AVP mRNA in the rat paraventricular nucleus (PVN) and supraoptic nucleus (SON), using in situ hybridization histochemistry. For nociceptive stimulation, formalin (5%) or saline was injected subcutaneously (s.c.) into the bilateral hind paws of rats. The expression of the nNOS gene in the PVN was significantly increased 2 and 6 h after s.c. injection of formalin in comparison with that in untreated and saline injected rats. The expression of the nNOS gene in the SON did not change in the untreated, saline- and formalin-injected rats. The AVP hnRNA in the PVN and SON was also significantly increased 15, 30 min and 2 h after s.c. injection of formalin, though AVP mRNA did not change at any time points that we studied. Plasma concentration of AVP was significantly increased 15 min after s.c. injection of formalin. These results suggest that NO in the PVN may be involved in nociceptive stimulation-induced neuroendocrine responses. PMID- 11474547 TI - Effect on urinary bladder function and arterial blood pressure of the activation of putative purine receptors in brainstem areas. AB - The effect on bladder function and arterial blood pressure of adenosine-5' triphosphate (ATP) and its synthetic analogue, alpha,beta-methylene ATP (alpha,beta-meATP) applied by microinjection to brainstem areas was assessed in the anaesthetised, paralysed and artificially ventilated female rat. Recordings of bladder pressure, changes in the pelvic nerve activity, arterial blood pressure and heart rate were evaluated. The purinergic drugs were microinjected into two brainstem areas the periaqueductal grey matter (PAG) and the area of the Barrington nucleus/locus coeruleus (LC) - only after electrical stimulation (50 Hz, 1 ms, 30-50 microA; n(PAG) = 17; n(LC) = 18) and the microinjection of glutamate (2 mM, pH 7.4+/-0.1; n(PAG) = 16; n(LC) = 16) had shown increases of bladder pressure and/or rate of bladder contractions and/or pelvic nerve activity at specific sites. Electrical and glutamate activation of PAG evoked an increase of arterial blood pressure. Microinjections of ATP (20 mM, pH 7.4+/-0.1; n(PAG) = 11; n(LC) = 11) and alpha,beta-meATP (2 mM, pH 7.4+/-0.1; n(PAG) = 10; n(LC) = 9) both evoked consistent increases of bladder pressure and/or pelvic nerve activity. Stimulation with ATP elicited a biphasic change of arterial blood pressure characterised by an increase followed by a decrease which was accompanied by a rise of heart rate. Microinjection of alpha,beta-meATP into PAG did not elicit a consistent response: a decrease of arterial blood pressure was evoked in five rats, while in two other rats an increase occurred. Electrical stimulation and glutamate activation of Barrington's nucleus/LC evoked an increase of arterial blood pressure, but a decrease was observed after microinjection of both ATP and alpha,beta-meATP. At some sites (n = 8) the effect of alpha,beta-meATP after a pre-injection at the same site of the P2 purino receptor antagonist, suramin (20 mM, pH 7.4+/-0.1) was smaller than the control. At three sites within PAG and two within LC located more medially to sites where an excitatory response had been observed, electrical stimulation evoked a small decrease or no change in bladder pressure. Following the stimulus, a rise in bladder pressure was preceded by an increase of pelvic nerve activity. A similar effect of glutamate was observed in one case. These data suggest that activation of P2 purine receptors in both PAG and Barrington's nucleus/LC is implicated in the neuronal mechanisms that generate patterns of activity in the parasympathetic innervation of the bladder and that purines also act at this level to modify sympathetic outflow to the cardiovascular system. PMID- 11474548 TI - Role of parabrachial nucleus in submandibular salivary secretion induced by bitter taste stimulation in rats. AB - When rats lick a bitter taste solution such as quinine-hydrochloride, they secrete profuse amounts of saliva. The salivation has a higher flow rate than that induced by other qualities of taste stimulation: sweet, salty, and sour. The present study is aimed to clarify the neural mechanism of the quinine-evoked salivation by means of behavioral, neuroanatomical, and electrophysiological experiments. Behaviorally, submandibular salivary secretion and rejection behavior (gaping) were observed in normal rats, as well as in rats chronically decerebrated at the precollicular level. In chronically decerebrate rats, these quinine-evoked reactions were strongly suppressed by destruction of the medial part of the parabrachial nucleus, including the so-called taste area, and ventral part of the parabrachial nucleus, including the pontine reticular formation. Neuroanatomical study using a retrograde tracer, Fluoro-gold, revealed that the neurons sending their axons to the superior salivatory nucleus, parasympathetic secretory center, were located mainly in the pontine reticular formation ventral to the parabrachial nucleus, not in the parabrachial taste area. Extracellular neural activity was recorded from the parabrachial region in decerebrate rats, and responsiveness to taste stimulation, jaw movements, and electrical stimulation of the superior salivatory nucleus was examined. Neurons responsive to both taste stimulation and antidromic stimulation of the superior salivatory nucleus were found in the pontine reticular formation ventral to the parabrachial nucleus, which responded well to quinine and HCl taste stimuli. Neurons in the parabrachial taste area could respond to four qualities of taste stimulation, but not to antidromic stimulation of the salivary center. These results suggest that aversive taste information from the parabrachial taste area reaches the salivary secretory center via the reticular formation ventral to the parabrachial nucleus. PMID- 11474549 TI - Comparison of aortic and carotid baroreflex stimulus-response characteristics in humans. AB - In order to characterize the stimulus-response relationships of the arterial, aortic, and carotid baroreflexes in mediating cardiac chronotropic function, we measured heart rate (HR) responses elicited by acute changes in mean arterial pressure (MAP) and carotid sinus pressure (CSP) in 11 healthy individuals. Arterial (aortic + carotid) baroreflex control of HR was quantified using ramped changes in MAP induced by bolus injection of phenylephrine (PE) and sodium nitroprusside (SN). To assess aortic-cardiac responses, neck pressure (NP) and suction (NS) were applied during PE and SN administration, respectively, to counter alterations in CSP thereby isolating the aortic baroreflex. Graded levels of NP and NS were delivered to the carotid sinus using a customized neck collar device to assess the carotid-cardiac baroreflex, independent of drug infusion. The operating characteristics of each reflex were determined from the logistic function of the elicited HR response to the induced change in MAP. The arterial pressures at which the threshold was located on the stimulus-response curves determined for the arterial, aortic and carotid baroreflexes were not significantly different (72+/-4, 67+/-3, and 72+/-4 mm Hg, respectively, P > 0.05). Similarly, the MAP at which the saturation of the reflex responses were elicited did not differ among the baroreflex arcs examined (98+/-3, 99+/-2, and 102+/-3 mm Hg, respectively). These data suggest that the baroreceptor populations studied operate over the same range of arterial pressures. This finding indicates each baroreflex functions as both an important anti-hypotensive and anti-hypertensive mechanism. In addition, this investigation describes a model of aortic baroreflex function in normal healthy humans, which may prove useful in identifying the origin of baroreflex dysfunction in disease- and training-induced conditions. PMID- 11474550 TI - Vascular and nociceptive effects of localized prolonged sympathetic blockade in human skin. AB - Supersensitivity to noradrenaline contributes to certain vascular disorders (e.g., hypertension) and chronic neuropathic pain conditions (e.g., complex regional pain syndrome). We aimed to develop a procedure for inducing adrenergic supersensitivity that could be used to investigate the role of catecholamines in these clinical conditions. In the first study, three doses of guanethidine were administered by iontophoresis to separate small patches of skin in the forearm of healthy human volunteers. Four to five hours later. the vasoconstrictor response to the adrenergic releasing agent tyramine was inhibited in a dose-dependent manner by iontophoretic pretreatment with guanethidine, indicating that guanethidine had depleted endogenous adrenergic stores. In a second study, guanethidine and saline were administered by iontophoresis four times over approximately 2 weeks at separate sites in the forearm. One to two days after the final pretreatment, vasoconstriction to the iontophoresis of a weak dose of noradrenaline was enhanced at sites pretreated with guanethidine. To investigate the effect of guanethidine pretreatment on thermal hyperalgesia. the experimental sites were sensitized to heat by the topical application of 0.6% capsaicin. Both before and after the application of capsaicin, the heat-pain threshold and heat pain ratings to suprathreshold stimulation were similar at sites pretreated for 2 weeks with guanethidine or saline. However, after the iontophoresis of noradrenaline, thermal hyperalgesia was greater at the guanethidine-pretreated site than the saline pretreated site. These observations indicate that prolonged depletion of adrenergic stores by guanethidine induces adrenergic supersensitivity in cutaneous vessels, and that adrenergic supersensitivity enhances thermal hyperalgesia in the presence of noradrenaline. PMID- 11474551 TI - Effect of tamsulosin hydrochloride on sympathetic hyperactivity in amyotrophic lateral sclerosis. AB - We assessed subclinical sympathetic hyperactivity in amyotrophic lateral sclerosis (ALS) patients, which might be followed by an autonomic spell leading to circulatory collapse, or sudden death as the disease progresses, and investigated the effect of tamsulosin hydrochloride (TSHC) on sympathetic hyperactivity. We measured the plasma norepinephrine (NE) concentrations of 41 ALS patients and 10 normal controls. TSHC, a selective alpha 1 blocker. was then administered to 10 ALS patients who had high plasma NE and to the 10 normal controls. Subsequent plasma NE change was evaluated for the possible alleviating effect of TSHC on subclinical sympathetic hyperactivity in ALS. Plasma NE was high in 20 of the ALS patients (48.8%), but had no relation to respiratory problems, which supports the previous speculation that plasma NE increases in ALS are not secondary to respiratory deficit, but reflect the primary pathomechanism of the disease. ALS patients showed a marked decrease in the NE concentration after TSHC administration, whereas there was no change in the controls. In conclusion, TSHC may be useful for suppressing central sympathetic hyperactivity, presumably the primary pathomechanism in ALS, and for preventing autonomic spells during the advanced stage of the disease. PMID- 11474552 TI - Decreased chaos and increased nonlinearity of heart rate time series in patients with panic disorder. AB - In this study, we investigated measures of nonlinear dynamics and chaos of heart rate time series in 30 normal control subjects and 36 age-matched patients with panic disorder in supine and standing postures. We obtained minimum embedding dimension (MED), largest Lyapunov exponent (LLE) and measures of nonlinearity (NL) of heart rate time series. MED quantifies system's complexity, LLE predictability and NL, deviation from linear processes. There was a significant increase in complexity (p < 0.00001), an increase in predictability (decreased chaos) (p < 0.00001) and an increase in nonlinearity (Snet GS) (p = 0.00001), especially in supine posture in patients with panic disorder. Increased NL score in supine posture may be due to a relative increase in cardiac sympathetic activity and an overall decrease in LLE may indicate an impaired cardiac autonomic flexibility in these patients due possibly to a decrease in cardiac vagal activity. These findings may further explain the reported higher incidence of cardiovascular mortality in patients with anxiety disorders. PMID- 11474553 TI - Recombinant hCG (OVIDREL) and recombinant interferon-beta1a (REBIF) (No. 13 in a series of articles to promote a better understanding of the use of genetic engineering). PMID- 11474554 TI - T (tail)-rhythm oscillators: principles of nervous control of the vasculature revealed? AB - This review focuses on the nervous control of the caudal ventral artery of the rat tail, and aims to convince the reader that sympathetic control of the vasculature can be mediated via neural oscillators intrinsic to the sympathetic nervous system. The definitive functional significance of these oscillators is unknown at present. However, it is expected that through dynamic relationships with modulating and driving inputs, such oscillators would permit graded vascular responses. PMID- 11474555 TI - Neurophysiological effects of recurrent laryngeal and thoracic vagus nerves on mediating the neurogenic inflammation of the trachea, bronchi, and esophagus of rats. AB - The present study aims to investigate the neurophysiological effects of recurrent laryngeal nerve and thoracic vagus nerve on the non-cholinergic regulation of neurogenic plasma extravasation of the rat trachea, bronchi, and esophagus. Through thoracotomy, three nerve components, the right thoracic vagal trunk, thoracic vagus nerve, and recurrent laryngeal nerve, were identified. The experiment was sequentially conducted in four steps. First, the individual nerve component was electrically stimulated and the induced inflammatory responses, as quantified by the area density of India ink-labelled blood vessels in the trachea, bronchial trees and esophagus, were compared. Second, we assessed the relative importance of medial and lateral side of the right thoracic vagus nerve in inducing the inflammatory responses by alternative stimulation of one side with simultaneous severance of the other side of this nerve. Third, we examined the effects of transection of the lateral half of the right thoracic vagus nerve on the degeneration of axon fibers located at the following three sites: the nerve segment proximal to cutting site, bronchial and esophageal nerve branches. Finally, we directly observed the inflammatory histopathology of the right lower trachea after stimulation of the medial half of the right thoracic vagus nerve with transection of its lateral half. In this study, we found that the right recurrent laryngeal nerve was predominant in mediating the neurogenic inflammatory responses of upper and dorsal portions of trachea, whereas the right thoracic vagus nerve was predominant in mediating those of the right lower ventral wall of trachea, right main bronchus, and right lobar bronchial trees. The axon fibers of the right thoracic vagus nerve responsible for mediating the neurogenic inflammatory responses of the right lower ventral trachea were mainly accumulated in the medial half, whereas those innervating the right main bronchus, right lobar bronchial trees, and lower esophagus were largely in the lateral half of this nerve. Transection of the lateral half of the right thoracic vagus nerve resulted in significant degeneration of myelinated fibers in its bronchial and esophageal nerve branches. Histopathological examination of the right lower trachea after electrical stimulation of the medial half of thoracic vagus nerve demonstrated the silver-stained leaky venules with accumulations of inflammatory cells. We thus concluded that afferent C-fibers to upper and dorsal portions of trachea were mainly from recurrent laryngeal nerve. In contrast, the neurogenic inflammatory responses of the right lower trachea were predominantly mediated by the medial half of the right thoracic vagus nerve, and those of the right main bronchus, bronchial trees and lower esophagus were largely by the lateral half of this nerve. PMID- 11474556 TI - Modulation of ACh-induced currents in rat adrenal chromaffin cells by ligands of alpha2 adrenergic and imidazoline receptors. AB - The aim of this study was to investigate the expression of the alpha2-adrenergic receptors in the adrenal medulla, and to examine the mechanism by which clonidine and related drugs inhibit acetylcholine (ACh)-induced whole-cell currents in adrenal chromaffin cells. Reverse transcription-polymerase chain reaction (RT PCR) performed on punches of rat adrenal medulla demonstrated expression of mRNA for the 2A-, alpha2B- and alpha2C-adrenergic receptors. Similar experiments conducted with tissue punches obtained from the adrenal cortex did not reveal expression of these receptor subtypes. Whole-cell currents were recorded in isolated chromaffin cells using the perforated-patch configuration. ACh (50 microM) evoked inward currents with a peak amplitude of 117.8+/-9.3 pA (n = 45; Vhol = -60 mV). The currents were inhibited in a dose-dependent manner (0.5-50 microM) by clonidine, UK 14,304 and rilmenidine (agonists of alpha2/imidazoline receptors), as well as by SKF 86466 and efaroxan (antagonists). Adrenaline and noradrenaline (50-100 microM) had no significant effect. Thus, although the adrenal medulla expresses mRNA for the alpha2-adrenergic receptors, the lack of agonist-antagonist specificity observed in our whole-cell recordings (in the absence of intracellular dialysis) provides additional evidence against the possibility that these inhibitory effects are mediated by classical alpha2 or imidazoline receptor interactions. PMID- 11474557 TI - Somatosensory nociceptive mechanical stimulation modulates systemic and mesenteric microvascular hemodynamics in anesthetized rats. AB - The effects of somatosensory nociceptive pinch stimulation of the hindpaw on mesenteric microvascular hemodynamics and systemic circulatory parameters were investigated in anesthetized rats using an intravital microscope-television system. Blood flow velocity in the terminal (18-40 microm in diameter) or precapillary (10-20 microm in diameter) arterioles of the mesentery was monitored by the dual sensor method developed by the authors. In the proximal terminal arterioles, blood flow velocity decreased substantially along with arteriolar constriction induced by pinching of the hindpaw for 30 s. In the distal terminal arterioles and precapillary arterioles, blood flow velocity increased after pinching. In the proximal terminal arterioles, the decrease of velocity in response to reflex vasoconstriction was abolished by intravenous injection of an alpha-blocker (phentolamine, 10 mg/kg). The increase in mesenteric precapillary arteriolar blood flow velocity (43+/-9%, p < 0.01) associated with the increase in mean arterial pressure (MAP) (22+/-1%, p < 0.01) was observed within a few seconds after the onset of the stimulation, and then the response in blood flow velocity returned to the baseline ahead of MAP response recovery after the end of the stimulus. These responses were diminished by alpha-adrenergic receptor blockade. The heart rate (HR) increase (4+/-1%, p < 0.01) induced by pinching was abolished by beta-adrenergicreceptor blockade (propranolol, 3 mg/kg, i.v.). There was a strong correlation between the increase in MAP and the decrease in renal blood flow measured by laser Doppler flowmeter (r = 0.87-0.98). Pinch stimulation of the rat hindpaw evoked changes in mesenteric arteriolar blood flow velocity that were mediated via the somato-sympathetic reflex vasoconstriction and the pressor response. PMID- 11474558 TI - Electrocardiographic changes during postnatal development in conscious swine with cardiac autonomic imbalance. AB - Using a conscious swine model, we studied the effects of different patterns of cardiac autonomic denervation on alterations of R-R and Q T intervals for 8 postnatal weeks. Newborn pigs were assigned randomly to four different groups: sham-operated controls (C), stellate ganglion ablation (SGX), either left (LSGX) or right (RSGX), and the right cardiac vagus nerve (RCVX) transection. The ECGs were recorded by telemetry while animals rested quietly or were judged behaviorally to be asleep. Analyses of the ECG included measurements of R-R and Q T intervals, as well as corrected Q-T intervals (QTc). Poincare plots were used to display age-related differences in R-R and Q-T intervals. For stellectomized animals, significantly prolonged R-R intervals were first observed at post surgical week 3 in the RSGX group and at week 5 in the LSGX group. Significantly prolonged QTc was found only in the RSGX group. In the RCVX group, shortened QTc and R-R intervals were noted at 6 and 7 weeks after denervation. Furthermore, three of six RSGX animals (50%) and one of four RCVX animals (25%) exhibited marked pauses in sinus rhythm that were unrelated to changes in heart rate or to sinus arrhythmia. These results in conscious animals support our hypothesis that abnormal autonomic innervation of the heart during maturation, e.g., withdrawal of vagal cardiac modulation or asymmetry of sympathetic innervation, impairs cardiac electrical stability. PMID- 11474559 TI - Stimulatory effect of isoferulic acid on alpha1A-adrenoceptor to increase glucose uptake into cultured myoblast C2C12 cell of mice. AB - In an attempt to elucidate the effect of isoferulic acid on alpha1-adrenoceptor (AR), the myoblast C2C12 cells of mice were employed to investigate the change of glucose uptake in the present study. Isoferulic acid enhanced the uptake of radioactive glucose into C2C12 cells in a concentration-dependent manner, which were abolished by pretreatment with prazosin. Effect of isoferulic acid on alpha1 AR was further characterized using the displacement of [3H]YM617 binding in C2C12 cells. The radioactive glucose uptake increasing action of isoferulic acid was abolished by tamsulosin or WB 4101 at concentration sufficient to block alpha1A adrenoceptor (alpha1A-AR) but it was not modified by chlorethylclonidine (CEC) at the concentration sufficient to abolish alpha1B-AR. An activation of alpha1A-AR by isoferulic acid in C2C12 cells can thus be considered. Pharmacological inhibition of phospholipase C (PLC) by U73312 resulted in a concentration dependent reduction of isoferulic acid-stimulated glucose uptake in C2C12 cells. This inhibition by U73112 was specific because the inactive congener, U73343, failed to modify the action of isoferulic acid. Also, chelerythrine and GF 109203X diminished the action of isoferulic acid at concentration sufficient to inhibit the activity of protein kinase C (PKC). The obtained data suggest that an activation of alpha1A-AR by isoferulic acid may increase the glucose uptake via PLC-PKC pathway in C2C12 cells. PMID- 11474560 TI - Non-dipper phenomenon in essential hypertension is related to blunted nocturnal rise and fall of sympatho-vagal nervous activity and progress in retinopathy. AB - Although the relation between the blunted nocturnal decline in blood pressure and target organ damages is well established, the mechanism underlying these results has not been clarified. We investigated the relationship among heart rate variability, nocturnal change in blood pressure and the severity of cardiac and extracardiac target organ damages caused by essential hypertension. We studied 52 Japanese inpatients with essential hypertension (24 men and 28 women; mean age, 49+/-3 years). After a stabilization period of 1 week, ambulatory blood pressure monitoring (ABPM) and 24-h ECG monitoring were performed and analyzed. The non dipper subjects were defined as those whose nocturnal decrease of mean BP was < 10% of daytime blood pressure (BP). The sex, age, body mass index. duration of hypertension, and 24-h BP were similar in dipper (n = 34) and non-dipper (n = 18) patients. The left ventricular mass index (LVMI) was significantly higher and the degree of hypertensive retinopathy was significantly worse in the non-dipper patients than that of the dipper patients. In the non-dipper patients, indexes of time-domain analysis such as the sum of differences between adjacent RR intervals (NNDrms), the number of pairs of adjacent RR intervals differing by more than 50 ms in the entire recording (RR 50) were significantly lower than that of the dipper patients. Additionally, as for spectral analysis, daytime low frequency/high frequency (LF/HF) was higher and nighttime high frequency (HF) was lower than that of the dipper patients. Independent predictors were the 24-h mean blood pressure (MBP) for left ventricular hypertrophy (LVH), nighttime systric BP (SBP) for progress in retinopathy and duration of hypertension for proteinuria. In conclusion, decrease in parasympathetic nervous function and increase in sympathetic nervous function may contribute to occurrence of non-dipper phenomenon, as well as progress in retinopathy. PMID- 11474561 TI - Myasthenia gravis with autoimmune autonomic neuropathy. AB - The autoantibodies that impair neuromuscular junction transmission in myasthenia gravis are specific for the nicotinic acetylcholine receptor (AChR) of muscle. Antibodies specific for AChRs in ganglionic neurons are found in a majority of patients with subacute autonomic neuropathy. Dysautonomia is not a recognized feature of myasthenia gravis, but there have been rare reports of myasthenia gravis coexisting with autonomic failure, usually in association with thymoma. Here we report seven patients who had myasthenia gravis with subacute autonomic failure. Their autonomic dysfunction ranged from isolated gastroparesis to severe panautonomic failure. Gastrointestinal dysmotility was a common feature. All had antibodies against muscle AChR, and three (all of whom had thymoma) had antibodies against neuronal ganglionic AChRs. In several patients, gastrointestinal function improved clinically after administration of an acetylcholinesterase inhibitor. These observations support a rare but definite clinical association between myasthenia gravis and autonomic failure and strengthen the concept that subacute autonomic neuropathy is an autoimmune disorder. PMID- 11474562 TI - [Topography of secondary dystonia lesions]. AB - Biological causes provoking dystonia can not be systematized, with the exception of the small group of levodopa-responsive dystonia. Therefore the pathophysiology of the dystonic syndrome can be approached by considering the site of the lesions. In 40 cases of uni or bilateral symptomatic dystonias, this site could be identified with CT Scan or MRI. Twenty-one were located in the striatum, six in the pallidum, seven in the thalamus, six in the midbrain. Each group is characterized by etiologic and clinical criteria, sometimes associated with abnormal movements. In the striatal group, the most important, dystonia was often associated by athetosis or choreoathetoid abnormal movements. In some cases, in children, lesions were vascular due to impairment of lenticulo-striatal arteries, often following cranial trauma. The pallidal lesions were usually provoked by metabolic or infectious agents. Most thalamic dystonias were of vascular origin, sometimes accompanied by myoclonia. Midbrain lesions were usually vascular with tremor. Athetosis occurred after striatal rarely after pallidal lesions. It is advisable not to assimilate dystonia and athetosis as both are simultaneously observed if the lesions are located in the striatum, rarely in the the pallidum but not in the midbrain. PMID- 11474563 TI - [A new concept to explain dysnatremia: the tonicity balance of entries and exits]. AB - Plasma sodium concentration, or natremia, results from three main factors: exchangeable sodium (Na+), exchangeable potassium (K+) and total body water (H2O). Its alterations often imply a change in cell volume. Understanding dysnatremias is essential for the treatment and prevention of hydromineral disorders. Extra-cellular fluid tonomoles consist almost exclusively of Na+ salts. Their dilution is the tonicity. K+ is an essential tonomole for intra cellular fluid tonicity. The balance between intra and extracellular tonicities depends on water movements and is responsible for changes in intra- and extracellular fluid volumes. Cell volume is therefore depending on the tonicity balance. A change in body tonicity (which is not osmolality) can be correctly and rapidly appreciated by measuring the (Na+ + K+) and H2O balances. Clinical cases emphasize the misleadings resulting from the free-water clearance calculation or the only measurements of urinary losses. They also demonstrate that tonicity balance provides indications for therapy whereas analyses based upon electrolyte free water do not. Intakes should be quantified with the same care than losses. The units used must be coherent to allow a quick and easy understanding at the bedside. Tonicity balance should be taught and Na+ + K+ and H2O balances should be routinely utilized by practitioners, dieticians and nurses in the concerned pediatrics, in particular intensive care, internal medicine, nephrology, pediatry and anesthesiology. PMID- 11474564 TI - [Prognosis of acute lymphoblastic leukemia in children. Results of the French protocol FRALLE 93]. AB - 1,120 children were included in protocol FRALLE 93 from june 1993 to september 1998. Disease Free Survival for the all protocol is 78% +/- 3 and overall survival 83% +/- 3. Various clinical and laboratory features at the time of diagnosis have been correlated with prognosis. They provide a potential mean to stratify patients into treatment subgroups according their relative risk of treatment failure. The identification of these prognostic factors has been an essential element in the design of current therapeutic trials. Prognostic characteristics of childhood ALL include: age, white blood cell count, tumor burden, cytogenetics (chromosome count and chromosomal translocation), immunophenotype and early response to treatment. Molecular biology has been the revolution of the last two decades permitting the cloning of the genes involved in the leukemic process. Finally the new molecular techniques allow a sensitive diagnostic approach to minimal residual disease (MRD). The better detection of MRD must allow a more rational basis for therapeutic intensification for a subset of poor responder patients. A decrease in therapy of very good responders can also be envisaged. PMID- 11474565 TI - [Results of surgical treatment of calcified aortic valve stenosis: report of a series of 4,129 interventions]. AB - Aortic stenosis (AS) is the most common lesion currently encountered among valvular heart disease, particularly in elderly people. Severe functional impairment and risk of sudden death explain that surgical treatment is largely accepted. We report a retrospective analysis of institutional experience with aortic valve replacement (AVR) for AS from 1971-1997 in 4,129 patients. Age ranged from 13 to 91 years (mean 68 +/- 10) and degenerative disease was largely predominant (86%). For AVR, mechanical prostheses were used in 2,054 patients (50.2%) and bioprostheses in 2,075 (48.8%) in elderly group. Coronary artery revascularization was associated in 670 patients (16%). Operative mortality was 7% (303 pts) and main cause was left ventricular failure (52%). Late results were studied with a maximum follow-up of 26 years. Total follow-up represents 21,533 pt-years. Late death occurred in 1,108 patients between 1 month and 24 years after operation (mean 6.6 years). Reoperation was necessary in 136 cases. Actuarial survival--including operative mortality--was 77% and 56% at 5 and 10 years. A large functional improvement was observed in the vast majority of patients, 73% being I or II subgroups of the NYHA classification. Incremental risk factors for death (immediate as well as late) were older age, preoperative functional status, emergency, presence of cardiac failure, coronary artery lesions and associated morbidity. The choice of valvular prosthesis remains controversial, but the results show that AVR is the procedure of choice for the vast majority of patients wtih significant aortic valve disease. PMID- 11474566 TI - [New progress and new tools for the study of molecular genetics in dyslipoproteinemia]. AB - More than three centuries after Mendel, at the era of electronic and computed information taking over the principle of information transmitted in discrete "packets" on the "internet", the sequence of the human genome is about to be completely released on public databases accessible on that very same internet. The gene, classically a virtual object, has become after several decades of intensive progress in cellular and molecular biology, a real object commonly manipulated and analyzed. More than fifty genes have been identified in the regulation of lipoprotein metabolism, giving rise to novel molecular pathophysiological bases for dyslipoproteinemia and beyond to other disorders related with lipid homeostasis. Dyslipoproteinemia, or disorders of lipoprotein metabolism commonly considered as lifestyle and age-related diseases, have now a molecular basis. Novel clinical entities no longer defined as "essential", but as molecular-based are progressively individualized. Novel tools for the diagnosis, prognosis or treatment have already modified the way these silent and frequent diseases are managed in clinical practice. In that respect, dyslipoproteinemia are among pioneer diseases in the medicine of the new millennium, which progressively evolves from a fact-based medicine to the individualized prevention of morbid events. PMID- 11474567 TI - [Role of anomalies of low density lipoproteins (LDL) in atherogenicity]. AB - Qualitative and quantitative anomalies of low-density lipoproteins (LDL) play a key role in the pathophysiology of atherosclerosis. Such anomalies are characteristics of the atherogenic dyslipidemias which occur most frequently, i.e. primary hypercholesterolemia of phenotype IIA (including familial hypercholesterolemia), combined hyperlipidemias (Type IIB) and hypertriglyceridemia (Type IV). An elevated concentration of circulating LDL occurs either as a result of hepatic overproduction of VLDL particles, the major precursors of LDL, or as a result of delayed catabolism, as occurs when there is a deficit of cellular LDL receptors (e.g. familial hypercholesterolemia), or as a combination of both. The major qualitative anomaly of LDL which results in elevated atherogenicity involves a predominance of small dense LDL, as seen in patients with premature coronary heart disease and equally in combined hyperlipidemia and in hypertriglyceridemia. The mechanism of the formation of these particles is complex and involves the concerted intravascular action of cholesteryl ester transfer protein (CETP), lipoprotein lipase (LPL) and hepatic lipase (HL) on triglyceride-rich precursors of dense LDL Lipid-lowering agents, such as fibrates and statins, act to reduce the atherogenicity of dense LDL by distinct mechanisms, which lead to normalisation of circulating LDL levels and/or to targeted reduction in dense particles of elevated atherogenicity. Indeed, such pharmacological probes have facilitated new insight into the molecular and cellular mechanisms which underlie each of the major forms of atherogenic dyslipidemia. PMID- 11474568 TI - [Clinical, epidemiologic, and biochemical findings on the reverse cholesterol transport (cholesterol-HDL)]. AB - Contrary to high plasma levels of low-density lipoprotein cholesterol (LDL-C), closely linked with coronary heart disease (CHD), high-density lipoprotein cholesterol (HDL-C) play an antiatherogenic role, through the reverse cholesterol transport from peripheral cells to the liver. New data in the pathophysiology of a rare genetic dyslipidemia, the Tangier disease, characterized by very low HDL-C levels and premature CHD, have shed light on this complex mechanism. In this disease, cholesterol efflux from peripheral cells is dramatically reduced, and this has been recently shown to be caused by mutations in an ATP-binding cassette transporter, which normally stimulates cholesterol efflux. Reverse cholesterol transport is therefore greatly decreased. Epidemiological data have revealed that 15% to 30% of coronary patients have low HDL-C levels. However, this is often combined with high triglycerides levels, and this association is frequently found in diabetic patients, especially prone to CHD. HDL-C has been repeatedly shown to be an inverse predictor of CHD. This has been enhanced by recent interventional studies (Veterans Affairs HDL Intervention Study, Bezafibrate Infarction Prevention Study) which have provided strong evidence that pharmacological increase of HDL-C, in combination with decrease in triglycerides level, reduces incidence of CHD. PMID- 11474569 TI - [Therapeutic and pleiotropic effects of statins in primary and secondary prevention of atherosclerosis]. AB - Strategy for management of patients displaying hyperlipidemia has changed since the first statin has been launched in France more than ten years ago. Following the first primary and secondary prevention studies conducted with hyperlipidemic subjects, results have been extended to patients with cholesterol levels within normal range both in primary (AFCAPS/TexCAPS) and secondary (CARE) prevention. Drug therapy has been simplified with targets tailored to the patient's risk. Despite these important results from intervention studies with statins, other crucial questions remain. Among these questions the first one concerns the possibility that cardiovascular benefits demonstrated with statins are also partly due to other (pleiotropic) effects. This was first hypothesised when statins were proven to be efficacious after short term treatments (one year in the regression studies). The second question is whether we can extend the benefit observed in populations included in published trials to primary prevention of stroke and to the groups less well represented in previous trials including women, elderly patients. The current trends and economic strains of every health care systems have focused on evidence-based medicine and cost-efficacy studies. Beyond LDL-cholesterol there is clear emergence of a role of triglyceride in cardiovascular disease as well as new data on pathophysiology of the so-called reverse cholesterol transport. Recent trials including patients with hypertriglyceridemia as well as a decline of the benefit observed in hypertriglyceridemic patients strongly suggest that the use of statins may have its own limitation. Finally, translating results of recent trials in clinical practice remains a challenge for treating physicians who face poor compliance and difficulties in implementing diet in patients. PMID- 11474570 TI - [New concepts on the mechanism of action of fibrates and therapeutic prospectives in atherosclerosis]. AB - Peroxisome Proliferator-Activated Receptors (PPARs) have been discovered 10 years ago as being orphan nuclear receptors. Three subtypes of PPAR(s) have been identified (alpha, gamma, delta). Activated PPARs bind to Peroxisome Proliferator Response Element which are localized in numerous gene promoters. PPAR(s) are activated by fatty acids and eicosanoids. PPAR-alpha activators (fibrates) improve plasma lipid levels and decrease CHD risk in patients with low HDL cholesterol (gemfibrozil). They also decrease atherogenesis (fenofibrate) in patients with type 2 diabetes. These drugs decrease atherogenic lipoprotein plasma levels such as VLDL and small dense LDL and they increase anti-atherogenic HDL, through increases in apo A-I and apo A-II synthesis. Furthermore, they induce overexpression in HDL receptors, such as SR-BI/CLA-1 and ABCA1 which are capable to increase cellular cholesterol efflux. Therefore, fibrates would reduce atherogenesis through their capacity to increase the "reverse cholesterol transport". Moreover, they would reduce vascular inflammation by repressing NF kappa B and AP-I transcriptional activity and they would reduce thrombosis risk by inhibiting tissue factor and fibrinogen synthesis. PMID- 11474571 TI - [Chondroid tissue]. AB - Nearly impossible to evidence with classical paraffin sections, chondroid tissue is regularly absent in the chapters of textbooks dealing with skeletal growth in which however it represents one of the major constituents. Microradiograph and methylene blue surface staining of thick undecalcified sections are the most suitable methods to reveal the presence of chondroid tissue. Moreover, all the methods used clearly indicate that this tissue is different from both calcified cartilage and woven bone or lamellar bone. The simultaneous presence, within the chondroid tissue matrix, of collagen type I, specific for bone tissue, and type II, specific for cartilage enables also to distinguish chondroid tissue from all the other calcified tissues. The presence of chondroid tissue in both fetal mandibular symphysis and in the sutural spaces of the skull strongly suggests that the same biomechanical stresses have the same consequences, i.e. the growth of the tongue separates the hemimandibles and the development of the brain has the same effect on the sutural areas. Experimental production of chondroid tissue is obtained in the space appeared between the bone fragments submitted to a continuous distraction. Finally, since the first cranial vault is also constituted by chondroid tissue islets, it has to be concluded that bone tissue is always secondary in its origin, i.e., after hyaline cartilage in endochondral ossification and after chondroid tissue in membranous ossification. PMID- 11474572 TI - [Perinatal medicine in the year 2000 seen by the anesthesiology-resuscitation specialist]. AB - Every year, 40.000 preterm babies are born in France, 10.000 of them being born before 33 weeks of gestation. They represent 1.2% of birth rate, but 50% of neonatal mortality. If IVF (in vitro fertilization) is the cause of less than 1% of all births, it is the cause of 7% of very preterm babies. Resuscitation of the very premature born infants has improved their survival rate but led to some adverse outcome. So, it is necessary to design a long term follow-up for children born from IVF, to evaluate the neonatal resuscitation and to improve treatment and care of children with developmental abnormalities. PMID- 11474573 TI - Autumnal IAP meeting. Sassari seminars of histopathology. Alghero, Italy, September 20-21, 2000. Proceedings and abstracts. PMID- 11474574 TI - Common germinal-center B-cell origin of the malignant cells in two composite lymphomas, involving classical Hodgkin's disease and either follicular lymphoma or B-CLL. AB - BACKGROUND: Classical Hodgkin's disease (HD) and B-cell non-Hodgkin lymphoma (NHL) occasionally occur in the same patient. Such composite lymphomas represent interesting models to study the pathogenesis of B-cell lymphomas and the relationship between HD and B-cell NHL. MATERIALS AND METHODS: We analyzed two composite lymphomas (a combination of classical HD with follicular lymphoma [FL] and a combination of classical HD with B-cell chronic lymphocytic leukemia [B CLL]) by micromanipulation of single cells from tissue sections and amplification of immunoglobulin V region genes for the clonal relationship of the tumor cells. RESULTS: In both cases, clonally related variable (V) genes with both shared as well as distinct somatic mutations were obtained from the two lymphomas, showing that in each of the cases the distinct tumor cells were members of a common germinal center (GC) B-cell clone. FL cells from two different lymph nodes of patient 1 showed a similar mutation pattern, suggesting that infiltration of these lymph nodes by tumor cells was not restricted to a particular FL cell or subclone. In the FL, a single cell was identified with a mutation signature indicating that premalignant cells can persist in the tissue. CONCLUSIONS: The cases presented here further underline the close relationship between HD and B cell NHL and the role of the GC in lymphomagenesis. Whereas the latter was already suggested for FL and HD, the present study indicates that also in the B CLL subset characterized by mutated Ig genes, important steps in malignant transformation happen in the GC, and that HRS cells can derive from CD5-positive B cells. PMID- 11474575 TI - The small GTPase Rac suppresses apoptosis caused by serum deprivation in fibroblasts. AB - BACKGROUND: The small GTPase Rac1 is a key signaling protein that mediates a number of important physiologic functions including the organization of the actin cytoskeleton, lipid metabolism, and gene transcription. Rac1 has also been implicated in oncogenic transformation. Expression of constitutively active Rac1 in Rat1 fibroblasts elicits serum- and anchorage-independent growth and causes tumorigenicity in nude mice. The signaling pathways that mediate the role of Rac in cell transformation remain to be identified. Here, we study the role of Rac in cell survival in the absence of serum. MATERIALS AND METHODS: The cell lines used in this study are Ratl fibroblasts that express constitutively active or dominant negative mutants of Rac1. We used long-term video time-lapse microscopy to analyze the effects of these Rac1 mutants on mitogenicity and apoptosis. RESULTS: We show that the increase in viability, which is stimulated by Rac1 in the absence of serum, is predominantly caused by an inhibition of apoptosis, with a minor increase in cell division. We also show that Rac1-stimulated cell viability in serum-starved cells is inhibited by chemical inhibition of phosphatidylinositol 3-kinase. CONCLUSIONS: Our observations indicate a role for Rac1 in survival signaling, possibly via activation of phosphatidylinositol 3 kinase. We propose that Rac1-stimulated cell survival may contribute to the role of Rac1 in serum-independent growth and cell transformation. PMID- 11474576 TI - Increased cytokine-induced cytotoxicity of pancreatic islet cells from transgenic mice expressing the Src-like tyrosine kinase GTK. AB - BACKGROUND: The loss of beta cells in type 1 diabetes may involve protein kinases because they control cell growth, differentiation, and survival. Previous studies have revealed that GTK, a Src-like protein tyrosine kinase expressed in beta cells (also named Bsk/Iyk), regulates multiple responses including growth and survival of rat insulinoma cells (RINm5F) and differentiation of neuronal PC12 cells. In the present study, we have generated a transgenic mouse expressing a kinase active GTK mutant (GTK-Y504F) under the control of the rat insulin I promoter to establish a role of GTK in beta cells. MATERIALS AND METHODS: Control and GTK-transgenic CBA mice were used for determination of in vivo glucose tolerance and the relative insulin-positive area. Isolated islets from both groups were cultured in the absence and presence of cytokines and insulin secretion, viability and protein expression were assessed. RESULTS: The beta-cell mass of GTK-transgenic mice was increased as a consequence of a larger pancreas and an increased relative beta-cell area. Islets isolated from the transgenic animals exhibited an enhanced glucose-induced insulin release and reduced viability in response to cytokines that could not be explained by higher levels of nitric oxide (NO) compared with control islets. Extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen-activated protein kinase (MAPK), c-Jun NH2-terminal kinase (JNK), and Akt were all activated by cytokines, but GTK-transgenic islets contained higher basal levels of phosphorylated ERK1/2 and lower basal levels of phosphorylated p38 compared with the control islets. The total amount of activated MAPKs was, however, higher in the cytokine-stimulated transgenic islets compared with the control islets due to increased levels of phospho-ERK1/2. Moreover, the proline-rich tyrosine kinase (PYK) 2 (also named RAFTK/CAK beta/CADTK) levels were elevated in response to a 24-hr exposure to cytokines in control islets but not in the GTK-transgenic islets. CONCLUSIONS: These results suggest that although GTK increases the beta-cell mass, it also enhances islet cell death in response to cytokines and may thus be involved in the beta-cell damage in type 1 diabetes. PMID- 11474577 TI - Islet endocrine-cell behavior from birth onward in mice with the nonobese diabetic genetic background. AB - BACKGROUND: Glucagon-producing alpha cells play a crucial role during the perinatal period. Because of their peri-islet localization near the early dendritic and macrophage cell infiltration, we thought it pertinent to investigate alpha cells in greater depth in nonobese diabetic (NOD) mice, a well recognized spontaneous model for human type I diabetes. MATERIALS AND METHODS: We determined alpha-cell distribution (glucagon immunohistochemistry and image analysis) and activity (real-time reverse transcriptase polymerase chain reaction [RT-PCR] and glucagon radioimmunoassay [RIA]), in relationship to glycemia in NOD and lymphocyte-deficient NODscid mice as compared to control mice (C57BL/6) from birth onward. RESULTS: NOD and NODscid mice, particularly at 1 day of age, had twice as many very small islets (<2,000 pixels) as C57BL/6 mice. During the postnatal period, the percentages of glucagon-positive areas in islets less than 2000 pixels were higher in NOD mice than C57BL/6; only a trend was found in NODscid. Pancreatic mRNA expression and glucagon content decreased in all strains at weaning. However, before weaning, pancreatic and blood glucagon levels were significantly lower in NOD and NODscid compared to C57BL/6 mice. Low basal nonfasting glycemia was observed in all strains before weaning with some strain differences: glycemia was significantly lower in NOD than C57BL/6, and higher in NODscid than NOD and C57BL/6. CONCLUSION: These data suggest that, before weaning, NOD and, to some extent NODscid pancreata contain more immature islets (as reflected by their small size and high percentages of glucagon-positive areas, concomitant with lower glucagon storage and basal secretion) than C57BL/6 pancreata. PMID- 11474578 TI - Expression and characterization of six mutations in the protoporphyrinogen oxidase gene among Finnish variegate porphyria patients. AB - BACKGROUND: Variegate porphyria (VP) is an inherited disorder of heme biosynthesis that results from a partial deficiency of protoporphyrinogen oxidase (PPOX). Patients with VP may experience acute neurovisceral attacks and cutaneous photosensitivity. To date we have characterized 109 VP patients representing 19 VP families in the Finnish population of 5 million, both biochemically and clinically. MATERIALS AND METHODS: Mutations were identified by direct sequencing of the patients' genomic DNA. The effect of the mutations was determined by sequencing the reverse transcriptase polymerase chain reaction (RT-PCR) product amplified from total RNA extracted from the patients' lymphoblast cell lines and expressing the mutations in E. coli and COS-1 cells. RESULTS: Of the six mutations identified in the PPOX gene, three mutations (IVS2-2a-->c, 338G-->C, and 470A-->4C) caused splicing defects, one produced a frameshift (78insC) and two mutations (R152C and L401F) caused amino acid substitutions. In RT-PCR, the IVS2-2a-->c mutation caused a retention of a 36-bp fragment in the 3' end of intron 2, the 338G-->C mutation caused an exon 4 deletion, and the 470A-->C mutation caused an exon 5 deletion with retention of a 19-bp fragment of the 3' end of intron 5. In both prokaryotic and eukaryotic expression systems, the PPOX activities of five mutants were decreased to 0-5% of the normal activity. CONCLUSIONS: This study describes five novel mutations and one earlier described major mutation among Finnish VP patients. All mutations produced detectable transcripts, but resulted in decreased PPOX activity confirming the causality of the mutations and the biochemical defects in these patients. PMID- 11474579 TI - Transcriptional regulation of the human iNOS gene by IL-1beta in endothelial cells. AB - BACKGROUND: Vascular endothelium participates in the control of vascular tone and function via the release of nitric oxide (NO) by the endothelial-type NO synthase (eNOS). Inducible NO synthase (iNOS) expression in endothelial cells occurs in many clinical conditions following induction by lipopolysaccharide or cytokines and generates large quantities of NO that result in endothelial cell activation and dysfunction. No information exists on the transcriptional regulation of the human iNOS gene (or that of other species) in endothelial cells. MATERIALS AND METHODS: We examined the transcriptional regulation of the human iNOS gene by interleukin-1beta (IL-1beta) in rat pulmonary microvascular endothelial cells (PVEC) by transient cotransfections of different iNOS-promoter constructs and cDNA of different transcription factors and regulatory proteins. RESULTS: The 1034/+88 bp iNOS promoter was strongly induced by IL-1beta, the regulatory elements for such induction being localized downstream of -205 bp. Cotransfection experiments with NF-kappaB isoforms, IkappaB isoforms, and IKK mutants suggested that the NF-kappaB site at -115/-106 bp is important, but not sufficient, for induction of iNOS promoter and that the role of NF-kappaB is partially independent of its binding site. C/EBP sites within the -205/+88 bp region were shown to be responsible, along with NF-kappaB site, for induction of iNOS promoter by IL-1beta. Overexpression of C/EBPalpha, C/EBPdelta, and liver enriched activator protein (LAP) activated the promoter, whereas overexpression of liver-enriched inhibitory protein (LIP) strongly suppressed it. C/EBPbeta (LAP and LIP isoforms) was constitutively present in PVEC and was induced (approximately 2-fold) by IL-1beta, whereas C/EBPdelta was not constitutively expressed but was strongly induced by IL-1beta. Both C/EBPbeta and C/EBPdelta participated in DNA-protein complex formation. CONCLUSION: Both NF-kappaB and C/EBP pathways are important for the transcriptional regulation of the human iNOS gene by IL-1beta in PVEC. PMID- 11474580 TI - Pulmonary inflammation induced by a recombinant Brugia malayi gamma-glutamyl transpeptidase homolog: involvement of humoral autoimmune responses. AB - BACKGROUND: A major allergen from the lymphatic filarial parasite Brugia malayi implicated in the pathogenesis of tropical pulmonary eosinophilia (TPE) has recently been cloned and identified as the homolog of the membrane-bound mammalian enzyme gamma-glutamyl transpeptidase (gamma-GT). Patients with acute TPE show autoreactive antibodies against endogenous gamma-GT from the pulmonary epithelium. MATERIALS AND METHODS: Recombinant B. malayi gamma-GT, alone or adsorbed to aluminium hydroxide (AL), was used in a BALB/c mouse model to analyze its antigenic/allergenic potential, its potential to induce pulmonary inflammation, and its capacity to induce autoreacting antibodies. RESULTS: Mice immunized with B. malayi gamma-GT showed significant levels of gamma-GT-specific IgG1, IgG2a, IgG3, IgA, IgE antibodies, and mild blood eosinophilia, even in the absence of adjuvant. Intranasal challenge with B. malayi gamma-GT induced peribronchial and perivascular inflammation characterized by a mixed infiltrate of lymphocytes, neutrophils, eosinophils, and macrophages. Both IL-4 and IFN gamma were detected in the peripheral blood and in the bronchoalveolar lavage fluid of immunized and intranasally challenged mice. Histological analysis of murine lungs using affinity-purified antibodies from mice immunized with the parasite's gamma-GT revealed the presence of autoimmune antibodies against pulmonary epithelium. Western blot analysis identified the 55 kDa heavy chain subunit of the murine gamma-GT as the target of autoreactive/crossreacting antibodies. CONCLUSION: Our data from the in vivo mouse model demonstrate the potent allergenicity/antigenicity of B. malayi gamma-GT, and its capacity to induce pulmonary inflammation upon intranasal challenge. This leads to breakdown of tolerance against endogenous murine gamma-GT. Thus, humoral autoimmunity against the airways epithelium may contribute to the pathogenesis of TPE. PMID- 11474582 TI - [History of the application of cold to food]. PMID- 11474581 TI - iNOS expression in dystrophinopathies can be reduced by somatic gene transfer of dystrophin or utrophin. AB - BACKGROUND: Nitric oxide (NO) is an inorganic gas produced by a family of NO synthase (NOS) proteins. The presence and the distribution of inducible-NOS (NOS II or iNOS), and NADPH-diaphorase (NADPH-d), a marker for NOS catalytic activity, were determined in muscle sections from control, DMD, and BMD patients. MATERIALS AND METHODS: NADPH-d reactivity, iNOS- and nNOS (NOS I)-immunolocalization were studied in muscles from mdx mice before and after somatic gene transfer of dystrophin or utrophin. RESULTS: In control patients, few fibers (<2%) demonstrated focal accumulation of iNOS in sarcolemma. In DMD patients, a strong iNOS immunoreactivity was observed in some necrotic muscle fibers as well as in some mononuclear cells, and regenerating muscle fibers had diffusely positive iNOS immunoreactivity. In DMD patients, NADPH-d reactivity was increased and mainly localized in regenerating muscle fibers. In mdx mice quadriceps, iNOS expression was mainly observed in regenerating muscle fibers, but not prior to 4 weeks postnatal, and was still present 8 weeks after birth. The expression of dystrophin and the overexpression of utrophin using adenovirus-mediated constructs reduced the number of iNOS-positive fibers in mdx quadriceps muscles. The correction of some pathology in mdx by dystrophin expression or utrophin overexpression was independent of the presence of nNOS. CONCLUSIONS: These results suggest that iNOS could play a role in the physiopathology of DMD and that the abnormal expression of iNOS could be corrected by gene therapy. PMID- 11474583 TI - [Technical problems associated with the cold chain]. AB - After having covered the current status of the cold chain in France, the author highlights the economic impact of refrigerated storage and processing involving refrigeration. Lose reduction is such that the return on investments is estimated as being roughly 3 years. In years to come, progress will be achieved in the field of plant design. Some information on design trends is provided. Operators in general, and carriers in particular, are faced with an increasingly complex regulatory framework leading to a competition imbalance. The current cold chain developmental trend is characterized by complex computer management of logistics: the flow of goods is modulated in order to meet the production to the distribution needs as cost effectively as possible. This logistic strategy enables management of downstream to upstream interfaces involving various operators. Finally, the author indicates the main causes of non-compliance with the cold chain, generally at retail and consumer levels. PMID- 11474584 TI - [Microbial growth and cold. Study of the particular case of Listeria monocytogenes]. AB - Some of micro-organisms are able to adapt themselves to hardest environmental conditions. So, about cold conditions, most of them can still grow at very low temperatures under 0 degree Celsius (nitrogen liquefied at -169 degrees C is the best way for preserving microbes). By another way some germs, qualified psychrotophic bacteria, grow quite easily between 0 degree C and +10 degrees C. Among these bacteria the author draw the attention to Listeria monocytogenes, a germ contaminating often foodstuffs and being responsible of deep diseases by eating food preserved in bad conditions (breaking off of refrigeration chain, bad use of domestic refrigerators). Recommendations are laid down to avoid these diseases, more particularly for frail consumers as immuno compromised adults. PMID- 11474585 TI - [Cold chain --the concept of traceability-- practical applications]. AB - Traceability makes it possible to monitor the properties of a foodstuff in general and its safety in particular. Based on the "refrigerated trio" concept developed by Andre Monvoisin, cold chain implementation conditions are examinated, with emphasis on the following three aspects, a safe product and early and continuous application of refrigeration. The TTT and PPP theories are presented. Following an overview of the French Academy of Medicine and the National Consumer Centre's recommendations, non compliance with the cold chain at household refrigerator level is discussed along with control equipment for refrigerators and products. Various indicators and integrators are presented and their described. Time-temperature integrators are examined in depth. In the conclusion, the authors stress the need to restore consumer confidence following recent foof-safety scares. PMID- 11474586 TI - [Factors affecting the temperature of domestic refrigerators]. AB - A survey was carried out in France in 1999 in order to know the air temperature in domestic refrigerators and the factors which may effect this temperature. Temperatures were recorded at three levels (top, middle and bottom of the refrigerator compartment). A questionnaire was filled to acquire the following information: characteristic of family (number of family members, age, profession, income...), characteristic of refrigerator (trade, type, age, temperature setting, refrigerating type...) and the use condition (room temperature, near by heat source, built in, door opening frequency...). The average temperature of the 119 surveyed refrigerators was 6.6 degrees C. Descriptive analysis and multi dimensional analysis of factors effecting refrigerator temperature were carried out. The classification tree and the segmentation confirm the influence of the use condition (frequency of door opening, temperature setting, near by heat source and built in). There is no direct effect of one factor but the combination of all of them. PMID- 11474587 TI - [Temperature and sensorial qualities of food]. AB - The pleasure of food-intake was emphasized by Brillat-Savarin in XIXo century. Beside pathogen bacterias, bad flavours caused by bacterial growth or enzymatic effects may happen in refrigerators with a mismanaged temperature. We have to distinguish between food-conservation and food-intake temperature. The ideal room temperature to appreciate a meal is about 22 degrees C with a damp of 60%. Relating to the four main flavours, salt and sweet are at their best at 18 degrees, bitter and sour at 8 degrees. All what is written before can be applied either in the case of sensorial analysis and meal. PMID- 11474588 TI - [Value of RHD fetal genotyping in the prevention of anti-D immunization]. AB - Anti-D prophylaxis is currently applied in France after birth of an RhD positive infant, after interruption of pregnancy and after some antenatal immunizing events (amniocentesis...). However this program does not cover all the prenatal exposures to fetal RhD antigen, and maternal Rh immunization continues to occur. DNA RhD genotyping of the fetus is now reliably performed on amniotic fluid, and pre diagnostic studies on fetal DNA extracted from maternal plasma are promising. The widespread use of fetal RhD genotyping on maternal blood would allow the antenatal administration of Rh immunoglobuline in all Rh negative patients bearing an Rh positive fetus, insofar as it would preclude exposing the other Rh negative patients to the above plasma derived and rather expensive blood product. PMID- 11474589 TI - [Microglia: origin and development]. AB - As suggested by Del Rio Ortega a long time ago, it is now widely accepted that microglia are the resident macrophages of the central nervous system. Microglia represent about 10% of the adult brain cell population. We have previously shown that the late embryonic and adult mouse brain contain potential microglial progenitors. We report here that microglial progenitors can be detected in neural folds from embryonic day 8. They originate from the yolk sac in which macrophage progenitors are found from embryonic day 7. We also report that the bulk of microglial cells (about 95%) appear during post-natal development. A major finding is that microglia arise by an intense in situ proliferation comparable to that of neural cells. Taken together, these results show that adult mouse microglia originate from cells migrating from the yolk sac and whose progeny actively proliferates in the brain during development. PMID- 11474590 TI - [Neurosteroids, their role in brain physiology: neurotrophicity, memory, aging...]. PMID- 11474591 TI - [Oocyte donation after the bioethics law. Medical, ethical and legal implications drawn form a series of 300 cases at the Tenon hospital]. AB - This is the perfect example of the problems which are the consequences of the actual medicine. We carried out an ovocyte donation study at the Tenon Hospital, in Paris, between 1994 and 1999 involving 177 cryopreserved thawed embryo transfers among 300 recipients. This study enables us to stress the ethical difficulties posed by the so called bioethical laws of 1994. Simultaneously two consequences became clearly evident: a paucity of donors, and the necessity to only transfer frozen embryos due to decree of 1996 upon sanitary security that imposes the quarantine of embryos for six months. On the other hand, the use of this method has yielded important new information regarding embryo implantation and the importance of ovocyte quality that is closely correlated to donor age. PMID- 11474592 TI - [Surgery of bronchial cancer after radiochemotherapy or chemotherapy. Risks and benefits]. AB - The purpose of this study was to evaluate the conditions and results of lung cancer surgery, following chemo and/or radiotherapy. This retrospective study included 69 patients treated from January 1990 to January 1998 for a primary lung cancer in whom surgery had been performed after induction treatment. Surgery had not been considered initially for the following reasons: N2 disease (n = 25), temporary functional impairment (n = 4); doubtful resectability (n = 40). The medical regimen resulted in combined radio-chemotherapy in 43 patients who received 2 to 4 sessions of chemotherapy (average = 2.9 +/- 0.8 sessions) and 43 +/- 8 Gy (20 to 60 Gy), or chemotherapy alone in 26 patients (3 +/- 0.7 sessions). Exploratory thoracotomy was performed in 4 patients (6%). There were 33 pneumonectomies, 1 bilobectomy, 23 lobectomies and 8 lung sparing resections. The in-hospital mortality was 9% (n = 6) from respiratory origin in all cases. There were 4 reoperations (6%): 3 for bronchial fistula and 1 for bleeding. Thirty five patients (51%) required blood transfusion (4.5 +/- 3.8 cell packs). The incidence of early and delayed bronchial fistula after pneumonectomy was 15%. Thirteen patients had a postoperative pneumonia (19%). The overall 5 years survival was 22% [19-32]. In the group of patients who had a complete resection, five-years survival for patients classified pathologically as N0 or N1 was 31% and, for those classified as N2, 8% (p = 0.19). Surgical management after induction chemo and/or radiotherapy of NSC lung cancer should be considered, in the absence of N2 disease, when a complete resection is achievable. However this surgery is associated with an increased risk. PMID- 11474593 TI - [Inherited human prion diseases]. AB - Familial prion diseases (familial Creutzfeldt-Jakob disease: Gerstmann-Straussler Scheinker disease, Fatal familial insomnia) are rare, but are also at present one of the most fascinating chapters of Neurology because of their double transmissibility. They are hereditary diseases of adults with a dominant autosomal transmission and an almost complete penetrance. They result most often of a point mutation of the gene of PrP with a consequent change in its primary sequence and conformation. Does the mutated PrP acquire a novel function or lose a still unknown function? At present there is no answer tho these question. The mutated PrP may sometimes be transmitted from man to animals. All PrP point mutations appear to have a noxious effect on neurons, but only some of them are transmissible. How mutated PrP acquires or does not acquire transmissibility may represent a fundamental progress in our understanding of prion diseases. PMID- 11474594 TI - [Familial articular chondrocalcinosis: study of an Alsatian family]. AB - Familial articular chondrocalcinosis is a chronic articular disease characterized by acute intermittent attacks of arthritis, presence of calcium pyrophosphate dihydrate crystal in synovial fluid, cartilage and periarticular soft tissue and by x rays calcium deposition in articular cartilage. A family originating from Alsace, with an autosomal dominant transmission has been studied. As in English and Argentinean families, a linkage to the short arm of chromosome 5p has been found. These results suggest that a defective gene at this location may be related to the chondrocalcinosis in these families. PMID- 11474595 TI - Standardized treatment of intra-articular calcaneal fractures using an oblique lateral incision and no bone graft. AB - BACKGROUND: The purpose of this study was to evaluate our protocol with the modified skin incision without bone graft in the treatment of intra-articular calcaneal fractures in a consecutive series. METHODS: Thirty-three displaced intra-articular calcaneal fractures operatively treated by a single surgeon were evaluated. Each fracture was managed according to a standardized protocol that included an oblique lateral incision and stable internal fixation, without supplemental bone graft. Fractures were classified according to two accepted criteria. Patients were evaluated with serial clinical and radiographic examinations. Final evaluation was made after at least a 2-year follow-up and was determined by the use of a functional outcome assessment questionnaire. RESULTS: Anatomic or near anatomic reductions were obtained in 97% of cases. All fractures healed without loss of initial reduction or collapse. Excellent or good results were achieved in 88% of non-worker's compensation patients and 27% of worker's compensation patients. Fractures with anatomic reductions were more likely to achieve a good or excellent result. Ninety-two percent of patients returned to the workforce. Complications included one superficial wound infection and five cases of peroneal tendinitis requiring hardware removal. One patient underwent subtalar fusion secondary to painful posttraumatic arthritis. CONCLUSIONS: The open reduction and internal fixation using our standardized protocol is an effective method of treatment for intra-articular calcaneal fractures. PMID- 11474596 TI - [Treatment of trophic ulcers in chronic venous insufficiency using bio transplants]. AB - The results of treatment of 66 patients with trophic ulcer in postthrombophlebitic syndrome, using biological lining, manufactured from mollusc of Cephalopoda genus owing properties of semipermeable biological membrane, were analyzed. Application of biogenic linings had promoted the results of patients treatment improvement due to quickening of epithelization and formation of ripe granular tissue. PMID- 11474597 TI - [Surgical treatment of aortal aneurysm]. AB - There were operated on 53 patients with aorta aneurysm. Operative mortality was 11.3%, but all 17 patients, in whom aminocapronic acid or aprotinin and synthetic prostheses were applied, survived after the operation. PMID- 11474598 TI - [Pulmonary resection of arteriovenous fistula]. AB - Arteriovenous fistula (AVF) is rare congenital pathology of the bronchopulmonary system vessels. The results of treatment of 12 patients with AVF were analyzed. In 2 (16.6%) patients Randu-Veber-Osler syndrome was diagnosed. In 10 observations the diagnosis was confirmed by the angiography data. All the patients were operated on. One (8.4%) patient died. PMID- 11474599 TI - [Destructive-perforative complications of an acute pancreatitis]. AB - There were examined 80 patients with fistula of various localization as complication of an acute pancreatitis. PMID- 11474600 TI - [Foreign bodies of biliary ducts as a cause of the obturative jaundice origin after cholecystectomy and other interventions]. AB - There were summarized the results of treatment of 17 patients with foreign bodies of biliary ducts which caused the obturation jaundice after cholecystectomy and other interventions. In 5 patients was applied the concealed rubber drain, in 12- polychlorvinyl one. All the patients were operated on. After the foreign bodies removal biliodigestive anastomosis was formed with external drainage of choledochus. After the operation one woman died due to complication by hepato renal insufficiency. PMID- 11474601 TI - [Modern aspects of treatment of colonic cancer, complicated by ileus. Part II. Treatment of colonic cancer, complicated by ileus]. AB - The results of treatment of 364 patients with colonic cancer, complicated by ileus. In 165 patients with partial ileus the radical operative interventions were performed, palliative--in 61 and symptomatic--in 83; with complete ileus--in 30.9 and 16 patients accordingly. Application of the interintestinal anastomoses formation methods, elaborated in the clinic, had permitted to perform primarily restorational operations in 81.9% of patients with partial ileus and in 61.5%- with complete one, obtaining satisfactory immediate results. Insufficiency of the duplicature anastomosis sutures had occurred in 5 (2.2%) of colonic cancer patients, suffering partial ileus. PMID- 11474602 TI - [Surgical treatment of the spleen damage in combined closed thoracoabdominal trauma]. AB - Experience of surgical treatment of 513 injured persons with closed thoracoabdominal trauma (CTAT) in an acute period was summarized. Splenic damage was revealed in 123 (23.9%) of patients. The CTAT structure was suggested as well as peculiarities of diagnosis, surgical tactics in splenic injury, complications and causes of mortality were analyzed. Mortality was 43.9%. PMID- 11474603 TI - [Treatment iatrogenic injury of rectum and colon]. AB - The results of treatment of 72 patients with iatrogenic injury of colon during performance of urological and gynaecological operations and endoscopic procedures, colonic investigations as well, were summarized. In 19 patients the injury was not revealed intraoperatively, causing the postoperative fecal peritonitis occurrence in 7 of them, abscess of small pelvis and the fecal fistula formation--in 12. PMID- 11474604 TI - [Cranio-abdominal injury]. AB - Cranioabdominal trauma (CAT) is one of the most severe forms of combined injury, in which the mortality rises up to 80%. Four principle factors in diagnosis and treatment of CAT are noted. The absence or noncomplete volume of the aid provided in the before hospitalization period causes unsatisfactory result of treatment in 30% of injured persons. Instrumental methods of investigation play the main role in CAT diagnosis. The informativity of laparocentesis in the closed abdominal trauma constitutes 98%. Operative intervention is performed under the support of antishock intensive therapy. When an urgent operation conduction is necessary it is performed by two teams of surgeons. PMID- 11474605 TI - [Reoperation in late terms after surgical treatment of morbid obesity]. AB - Experience of the reoperations performance after jejunoileoshunting and gastric volume reducing operations for morbid obesity (MO) was demonstrated on. They may serve as a concluding phase of MO surgical treatment (various variants of dermatolipectomy) or as a reconstructive-restorational operations, directed on correction of various pathological syndromes. PMID- 11474606 TI - [The state of the pulmonary surfactant system in patients with the acute pulmonary damage syndrome]. AB - There were examined the surface-active properties and phospholipidious contents of the breathed out air condensate and the general respiratory compliance (C) in 21 patients with an acute pulmonary damage syndrome (APDS), caused by pancreonecrosis, peritonitis, polytrauma, sepsis. The lowering of the surface activity, reducing of phosphatidylcholine content and raising of phosphatidylethanolamine level, significant C lowering were noted. Such changes were more pronounced in patients, which have died subsequently. This testifies the profound inhibition of the pulmonary surfactant system activity in patients with APDS. It is expedient to apply the exogenic surfactant preparations for the treatment of patients with APDS. PMID- 11474607 TI - [Non-clostridial epi-fascia phlegmon]. AB - Complex method of surgical treatment of epifascial phlegmon is elaborated. The flaps tailoring and dividing in accordance to anatomic site and its blood supply permits to do radical cleansing and sanitation of focus with mixture of hydrogen peroxide and spirituous solution of iodine preserving the viable skin maximally. In contrast to conventional "stripe" incisions, application of the proposed method permits to preserve the wound from significant lympho--and plasma losses, causing the wound exhaustion, secures the possibility of visual control of the necrotic process course. The wound coverage with cutaneo-subcutaneous flap of the full value assists reducing of the pain intensity. The flap fixation prevents its shortening, that's why good cosmetic effect is noted after the wound healing concludes. Complex program of the necrotic fascitis treatment proposed had permitted to reduce the hospitalization duration of patients twice and to lower the mortality rate more than in 3 times. PMID- 11474608 TI - [Noncomplicated and complicated healing of wound in patients after hemorrhoidectomy]. AB - Complicated and noncomplicated forms of rehabilitational process course after hemorrhoidectomy are delineaed. There were studied up an organism reactivity, the peroxidal oxidation of lipids intensity, the dynamics of the biogenic amines contents. The dependence of the inflammation severity in the wound from severity of the organism stress-reaction on operative intervention was established. In patients with normal reactivity the noncomplicated healing of operative wound was observed. In patients with lowered or enhanced reactivity there were observed the reduction of the antioxidant system limiting activity, disbalance of pro- and antioxidation systems, the biogenic amines dynamics change with long drawn-out course of inflammatory reaction and with complicated healing of operative wound. PMID- 11474609 TI - [Application of auto-dermal hernioplasty for treatment of postoperative abdominal hernia]. AB - The results of surgical treatment of 98 patients with postoperative abdominal hernia, in whom various methods of autodermal hernioplasty were applied, were summarized. In 33 patients for the hernial defect closure deepithelized cutaneous strip was used, in 22--free cutaneous flap, in 12--the defect was closed with the help of free armourized autodermotransplant, in 10 patients--with free cutaneous flap in combination with lavsan mesh. Recurrency after application of cutaneous strip had occurred in 41% of patients, of cutaneous flap--in 36.3%, armourized flap--in 23.1%, after the combined plasty--in 20%. Insufficient blood circulation in transplant is the cause of high recurrency rate after doing the plasty using free flaps. To eliminate this shortage and to improve the results of surgical treatment in 20 patients there was applied original method of autodermal hernioplasty using deepithelized cutaneous flap on two nourishing pedicles. Recurrency had occurred in one patient during follow-up from 1 to 5 years. PMID- 11474610 TI - [Results of surgical treatment of postoperative abdominal hernia]. AB - There were examined 525 patients with postoperative abdominal hernia, in 47.3% of them big, vast and giant hernia was revealed. There were operated 436 patients using local tissues with duplicature formation--according to Mayo, Sapezhko, Napalkov and Yanov method. PMID- 11474611 TI - [Urgent interventions for invasive thyroidal carcinoma]. AB - There was summarized the experience of treatment of the thyroid gland cancer, invading trachea (larynx) and other neighboring structures of the neck in 42 patients, whose state was estimated as severe one and neoplastic process--as nonoperable. Optimal surgical methods were applied depending on clinical signs of thyroidal carcinoma. In 36 patients the palliative resection of thyroid gland (in 6--with tracheostomy addition) was done, in 6--intervention was restricted by tracheostomy formation. The method of tracheostomy formation in cases of massive spreading of tumor along trachea is adduced. PMID- 11474612 TI - [Wound healing effect of preparation vermilat for injections in experiment]. AB - Results of studying of the woundhealing effect of preparation vermilat for injections, which constitutes the extract of polypeptide origin, were adduced. The skin wound was done to the Wistar line rats and than the preparation in 0.1 mg dose of dry substance was injected. The preparation had caused pronounced effect on regeneration of tissues. The expediency of application of the preparation in the wound healing process during its second and third stages was proved. PMID- 11474613 TI - [Thromboembolism of pulmonary artery]. PMID- 11474614 TI - [Closed pancreatic injury]. PMID- 11474615 TI - [Influence of amino acids on pancreatic functional activity]. PMID- 11474616 TI - [Chronic venous insufficiency of the lower extremities in patients with post thrombophlebitic syndrome]. AB - The results of examination and treatment of 734 patients with postthrombophlebitic syndrome of lower extremities were adduced. The pathogenesis diagram for chronic venous insufficiency was proposed. The possibilities of the regional blood flow correction using reconstructive operations on veins were shown. In 73% of patients good result in immediate postoperative period was noted and in late follow-up period--in 64%. PMID- 11474617 TI - [Surgical correction of the intrathoracic obstruction of respiratory tracts in the presence of the pulmonary artery "loop" in children]. PMID- 11474618 TI - [Surgical errors during colostomy in children with Hirschsprung's disease]. PMID- 11474619 TI - [Method of exclusion of intestinal fistula]. PMID- 11474620 TI - [The delayed sternum closure after operation of arterial over-switching in D transposition of the main vessels]. PMID- 11474621 TI - [Surgical strategies in cardiac wound treatment]. PMID- 11474622 TI - [Observation of numerous gun shots of the abdomen and retroperitoneal injuries]. PMID- 11474623 TI - [Application of video-thoracoscopy in differential diagnosis of mediastinal tumors]. PMID- 11474624 TI - [Multiple cholangiogenic hepatic abscesses in a patient with diffuse purulent peritonitis and septic shock]. PMID- 11474625 TI - [The gallbladder agenesis]. PMID- 11474626 TI - [Arterial directions of the hand's venous flow]. AB - In 9 patients arterialization of venous bed of manus was performed. The operation technique and results of treatment are adduced. The modern approach to the method of treatment choice had permitted to achieve positive results in 88.9% of operated patients. PMID- 11474627 TI - [XX Consensus Conference in Resuscitation and Emergency Medicine. Corticotherapy in septic shock and respiratory distress syndrome]. PMID- 11474628 TI - CDC says there are ways to reduce enteric pathogen transmission in swimming pools. PMID- 11474629 TI - HHS releases first guide specifically addressing medical treatment of women with HIV. PMID- 11474630 TI - Unexplained Deaths and Critical Illnesses Project (UNEX). PMID- 11474631 TI - Notice to readers: deferral of routine booster doses of tetanus and diphtheria toxoids for adolescents and adults. PMID- 11474632 TI - Susceptibility of a variety of clinical isolates to linezolid: a European inter country comparison. AB - Using standardized in vitro susceptibility tests, 3382 bacteria recently isolated from skin, blood or respiratory tract infections were analysed for their susceptibility to linezolid, a new oxazolidinone, and a number of comparator antibacterial agents. Isolates originated in France, Italy, Germany, Spain, Sweden, The Netherlands and the UK. Laboratories in each country independently conducted broth microdilution susceptibility tests using NCCLS methods and epsilonometry (Etest). Isolates of Gram-positive cocci tested in each laboratory included methicillin-susceptible and -resistant Staphylococcus aureus, methicillin-susceptible and -resistant Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus pneumoniae and Enterococcus spp. Isolates of Moraxella catarrhalis and Haemophilus influenzae were also included. Where appropriate, comparator drugs (oxacillin, vancomycin, gentamicin, co-amoxiclav, ciprofloxacin, erythromycin, penicillin G, clindamycin and ampicillin) were also tested. Linezolid demonstrated excellent activity against all of the Gram-positive cocci with MIC50s ranging from 0.5 to 4 mg/L. The drug demonstrated only modest activity against M. catarrhalis and H. influenzae with MIC50s ranging from 4 to 16 mg/L. PMID- 11474633 TI - Efficacy and safety of gemifloxacin in the treatment of community-acquired pneumonia: a randomized, double-blind comparison with trovafloxacin. AB - This multicentre, randomized, double blind, parallel group study compared the efficacy and safety of gemifloxacin (320 mg once daily) with trovafloxacin (200 mg once daily) in 571 patients with community-acquired pneumonia (CAP). Although treatment was given routinely for 7 days it could be extended to 14 days; two thirds of patients were treated for 7 days. High clinical success rates were noted at follow-up in the per-protocol population in both the gemifloxacin group (95.8%) and the trovafloxacin group (93.6%), non-inferiority with 95% CI. In the intent-to-treat population, the clinical success rate at follow-up was significantly superior for gemifloxacin (87.6%) compared with trovafloxacin (81.1%; 95% CI 0.5, 12.4). The pathogens identified most commonly at presentation were Mycoplasma pneumoniae and Streptococcus pneumoniae. Gemifloxacin eradicated 100% of S. pneumoniae. One bacteraemic isolate of S. pneumoniae was associated with clinical failure in the trovafloxacin group (MIC of trovafloxacin 8 mg/L). Gemifloxacin was well tolerated and the incidence of transient liver function abnormalities was very low. Gemifloxacin is an effective and well-tolerated treatment for patients with CAP. PMID- 11474634 TI - Health plans, insurers unsure about likely effect of Gramm-Leach-Bliley. PMID- 11474635 TI - Compensation monitor. Incentives make the difference. PMID- 11474636 TI - Managed care outlook. 'Quality of care' in eye of beholder. PMID- 11474637 TI - Daily changes in beta-adrenergic sensitivity of rat submandibular gland. Correlation with beta-adrenoceptor rhythm. AB - Neurons from the superior cervical ganglia (SCG) innervate the submandibular gland and release noradrenaline during the dark phase of the daily photoperiod. Since in the pineal, another structure innervated by sympathetic neurons, nocturnal activation of the SCG is associated with beta-adrenergic sub- and super sensitivity rhythms, the possible existence of similar phenomena in the rat submandibular gland was assessed. Wistar female rats, kept on a 14:10 light/dark cycle (light from 06:00 to 20:00 h), were sacrificed at 09:00, 14:00, 20:00, 24:00 and 04:00 h. beta-Adrenoceptors were studied by 3H-dihydroalprenolol binding to membrane preparations. The equilibrium dissociation constant (Kd) did not change as a function of time while significant daily variations in maximal binding values (Bmax) were observed with a peak at 20:00 h. Changes in Bmax correlated with a high response of adenylate cyclase to isoproterenol. In addition, when the response in salivary flow to isoproterenol was measured. a shift to the left (about 1 logarithmic unit) in dose-response curves was observed at 19:00-20:00 has compared to 08:00-09:00 h. These daily variations in isoproterenol responsiveness seem not to depend on the pattern of eating since a 24-h starvation or a nocturnal starvation for 16-18 days did not abolish the morning-evening differences in the salivary flow response to isoproterenol. Rather, the results suggest that the daily variations in isoproterenol response correlate with beta-adrenergic super- and sub-sensitivity phenomena associated with the circadian release of noradrenaline from SCG neurons. PMID- 11474638 TI - The sweating apparatus in growth hormone deficiency, following treatment with r hGH and in acromegaly. AB - Adult growth hormone deficient patients are known to exhibit reduced sweating and their ability to thermoregulate is diminished. Treatment of these patients with recombinant human growth hormone (r-hGH) is claimed to reverse these abnormalities. We have investigated this claim, as well as the mechanism underlying these altered sweating responses in GH-deficient patients as part of a placebo-controlled study on the effects of 6-12 months r-hGH therapy. Skin biopsies were obtained from these subjects and changes in morphology and innervation parameters for the eccrine sweat glands were examined. These included histochemistry for acetylcholinesterase (AChE) and immunohistochemistry for the neuropeptide vasoactive intestinal polypeptide (VIP) and for PGP9.5, a general neuronal marker. Sweat gland acinar size and periacinar innervation were measured by computerised image analysis. The patients underwent pilocarpine iontophoresis sweat rate tests and their serum insulin-like growth factor 1 (IGF-1) levels were assessed. Since active acromegaly involves excess GH secretion and hyperhidrosis, skin biopsies and sweat tests were also carried out on a group of these patients, as well as on control subjects. We have demonstrated a sweating defect in adult GH-deficiency which is accompanied by a reduction in AChE and VIP levels in the nerve supply to sweat glands. Following r-hGH therapy, an increase in AChE and VIP staining is seen in the sudomotor nerves accompanied by restoration of sweat rates and serum IGF-1 levels. Hence, normalization of sweat gland function includes recovery of sudomotor synapse constituents. A trophic effect of GH on sweat gland epithelium and/or on the associated nerves is proposed, supported by the observation that in acromegaly the size of sweat gland acini and the density of innervation to the sweat glands was greater than in controls. PMID- 11474639 TI - Anatomic relationships of the human nucleus paragigantocellularis lateralis: a DiI labeling study. AB - The nucleus paragigantocellularis lateralis (PGL) is located in the rostral ventrolateral medulla (RVLM), a brainstem region that regulates homeostatic functions, such as blood pressure and cardiovascular reflexes, respiration. central chemosensitivity and pain. In the present study, we examined anatomic relationships of the human nucleus paragigantocellularis lateralis using a bidirectional lipophilic fluorescent tracer, 1,1'-dioctadecyl-3,3.3',3' tetramethylindocarbocyanine perchlorate (DiI), in nine postmortem human fetal midgestational brainstems. The areas which were labeled by diffusion of DiI from the nucleus paragigantocellularis lateralis included the arcuate nucleus (ARC) of the medulla, caudal raphe (nucleus raphe obscurus and pallidus), hilum and amiculum of the inferior olive, bilateral "reticular formation" (including the nucleus paragigantocellularis lateralis, nucleus gigantocellular-is and the intermediate reticular zone (IRZ)). vestibular and cochlear nuclei, cells and fibers at the floor of the fourth ventricle with morphologic features of tanycytes, parabrachial nuclei (PBN), medial lemniscus, lateral lemniscus, inferior cerebellar peduncle and cerebellar white matter, central tegmental tract, and the capsule of the red nucleus. This pattern of DiI labeling bears many similarities with the pattern of connections of the nucleus paragigantocellularis lateralis previously demonstrated by tract-tracing methods in experimental animals, and is consistent with the role of the nucleus paragigantocellularis lateralis in central regulation of homeostatic functions. In contrast to the animal studies, however, we did not demonstrate connections of the nucleus paragigantocellularis lateralis with the nucleus of the tractus solitarius (nTS) (only connections with the rostral subdivision were examined), locus coeruleus, or the periaqueductal gray (PAG) in the human midgestational brainstem. In our previous studies, six medullary areas showed reduced serotonin receptor binding in a subset of victims of sudden infant death syndrome (SIDS). The present study demonstrated DiI labeling in all of these six areas, suggesting that they are interconnected. PMID- 11474640 TI - Choline acetyltransferase activity parallels the pressure gradient in the feline pharyngo-esophageal region. AB - Pressures generated by the pharyngeal constrictor muscles and proximal esophagus involve acetylcholine-induced muscle contractions. We hypothesized that the pharyngo-esophageal pressure gradient is related to choline acetyltransferase activity. In nine anesthetized cats, hypopharyngeal pressure and proximal esophageal pressure were recorded with a solid state transducer assembly. Enzymatic activities in the thyropharyngeus, cricopharyngeus, and proximal esophageal muscles were measured. Hypopharyngeal pressure was higher than the proximal esophagus (p < 0.01), and choline acetyltransferase activity was higher in the cricopharyngeus compared to the proximal esophagus ( p < 0.05). The pressure gradient between the hypopharynx and proximal esophagus may be influenced by the activity of choline acetyltransferase. PMID- 11474641 TI - Cardiac autonomic denervation and functional response to neurotoxins during acute experimental Chagas' disease in rats. AB - Severe cardiac autonomic denervation occurs in the acute Chagas' disease in rats. The present study aims at verifying whether this denervation was accompanied by impairment of heart function. Scorpionic (Tityus serrulatus) crude venom was used for neurotransmitter release in isolated hearts (Langendorff's preparation). In control hearts, the venom induced significant bradycardia followed by tachycardia. In infected animals, despite the severe (sympathetic) or moderate (parasympathetic) cardiac denervation, the venom provoked similar bradycardia but the tachycardia was higher. The hearts of infected animals beat at significantly lower rate. Atropine prevented this lower rate. Our results demonstrated sympathetic dysfunction during the acute phase of Trypanosoma cruzi infection in rats, the parasympathetic function being spared. PMID- 11474642 TI - Adrenergic vasoconstrictor activity in the cerebral circulation after inhibition of nitric oxide synthesis in conscious goats. AB - The interaction between nitric oxide (NO) and adrenergic activity in the cerebral circulation was studied using conscious goats, where blood flow to one brain hemisphere (cerebral blood flow) was electromagnetically measured, and the effects of phentolamine and hexamethonium on cerebrovascular resistance were evaluated before (control) and after inhibition of NO synthesis with NW-nitro-L arginine methyl ester (L-NAME). L-NAME (12 goats, 40 mg kg(-1) administered i.v.) reduced cerebral blood flow from 62 +/- 3 to 44 +/- 2 ml min(-1), increased mean systemic arterial pressure from 100 +/- 3 to 126 +/- 4 mm Hg, decreased heart rate from 79 +/- 5 to 50 +/- 4 beats min(-1) and increased cerebrovascular resistance from 1.63 +/- 0.08 to 2.91 +/- 0.016 mm Hg ml(-1)min(-1) (all P < 0.01). These hemodynamic variables normalized 48-72 h after L-NAME administration. Phentolamine (six goats, 1 mg), injected into the cerebral circulation. increased cerebral blood flow without changing systenic arterial pressure, but its cerebrovascular effects were augmented for about 24 h after L NAME. The decrements in cerebrovascular resistance induced by phentolamine, in mm Hg ml(-1) min(-1), were: under control, 0.42 +/- 0.05; immediately after L-NAME, 1.38 +/- 0.09 (P < 0.01 compared with control); by about 24 h after L-NAME, 0.71 +/- 0.09 (P < 0.05 compared with control); and by about 48 h after L-NAME, 0.40 +/- 0.07 (P > 0.05 compared with control). Hexamethonium (six goats, 0.5-1 mg kg( 1) min(-1) i.v.) decreased mean systemic arterial pressure to about 75 mm Hg and caused tachycardia similarly before and after L-NAME, but the decrements in cerebrovascular resistance were augmented for about 24 h after L-NAME. The decrements in cerebrovascular resistance induced by hexamethonium, in mm Hg ml( 1).min(-1), were: under control. 0.61 +/- 0.09, immediately after L-NAME, 1.33 +/ 0.16 (P < 0.01 compared with control); by about 24 h after L-NAME, 1.18 +/- 0.10 (P < 0.01 compared with control): and by about 48 h after L-NAME, 0.99 +/- 0.10 (P > 0.05 compared with control). Therefore, these results suggest that adrenergic vasoconstrictor tone in cerebral vasculature may be augmented after inhibition of NO synthesis, and that this increment may contribute to the reduction of cerebral blood flow after inhibition of NO formation. PMID- 11474643 TI - Nonlinear dynamics applied to blood pressure control. AB - Hypertension is a very frequent disease, known to trigger a range of severe cardiovascular problems. The elucidation of its pathophysiology requires investigation of the mechanisms responsible for the maintenance of blood pressure in the normal system, and their possible failure in hypertension. Some of these control mechanisms display nonlinear features, indicating that the blood pressure signal might be characterized by nonlinear dynamics. Our aim was thus to investigate the nonlinear properties of the blood pressure signal under normal conditions, and in a cardiovascular system prone to hypertension. Blood pressure was investigated in young spontaneously hypertensive rats (SHR), versus their age matched normotensive progenitors (WKY). The correlation dimension was computed as quantification of blood pressure control complexity. The parameters required for the calculation procedure of the correlation dimension were carefully determined. The results were tested with surrogate data statistics. assuming linear autocorrelated Gaussian noise as the null hypothesis. Non-integer correlation dimension values were found in both strains, with lower values for SHR than for WKY, in particular following alpha-blockade. In all cases, a statistically significant difference was found between the real and surrogate data. These results show that the nonlinear dynamics parameter D, can be used to detect differences in BP control between prehypertensive SHR and WKY rats as early as 6 7 weeks after birth. PMID- 11474644 TI - Dendritic hypertrophy of Stach type VI neurons within experimentally altered ileum of pigs. AB - Myenteric neurons were investigated morphometrically to answer the question if type-specific somal hypertrophy of type VI neurons in mechanically stressed ileum of pigs, which was known from an earlier study, is correlated with an increased dendritic arborization, that is, with dendritic hypertrophy. Muscular hypertrophy was induced in the ileum of two juvenile pigs by narrowing the gut circumference (mechanical stenosis) and by reversing a loop of ileum which results in an antiperistaltic segment (functional stenosis), respectively. After a survival time of 6 weeks, wholemounts from the pre- and poststenotic ileal regions, from the antiperistaltic segment as well as from an age matched control animal, were silver impregnated. Dendritic parameters of Stach types IV and VI neurons were recorded using a computer-aided morphometric program and analysed statistically. Type IV neurons showed no change of dendritic parameters, neither within control nor within stenosed ileal segments. In contrast, the type VI neurons displayed increased dendritic parameters within zones of muscular hypertrophy such as total dendritic length, numbers of dendrites, of dendritic branching points and of dendritic endings. We suggest that type VI neurons may participate as descending nitrergic interneurons or motorneurons in the control of muscular function, thus, undergoing plastic changes in case of experimental muscular hypertrophy. Type IV neurons which are involved in the regulation of mucosal processes were not affected by muscular hypertrophy. PMID- 11474645 TI - Rostral ventrolateral medullary opioid receptor subtypes in the inhibitory effect of electroacupuncture on reflex autonomic response in cats. AB - Electroacupuncture (EA) is used in traditional Chinese medicine to treat arrhythmias, hypertension and myocardial ischemia. Our previous work suggests that the inhibitory effect of EA on the pressor reflex induced by bradykinin (BK) applied to the gallbladder is due, in part, to the activation of opioid receptors, most likely located in the rostral ventrolateral medulla (rVLM). However, specific opioid receptor subtypes, and hence the neurotransmitters. responsible for this inhibition are unknown. Therefore, in anesthetized cats, BK (10 microg/ml) was applied to the gallbladder to induce transient reflex increases in arterial blood pressure (BP). EA (1-2 mA, 5 Hz, 0.5 ms pulses) was delivered through acupuncture needles inserted bilaterally into Neiguan and Jianshi acupoints on forelimbs, overlying the median nerves. EA attenuated the BK induced pressor response by 39%. Opioid receptor subtype antagonists or agonists were microinjected unilaterally into the rVLM. The mu- and delta-receptor antagonists CTOP and ICI 174,864, respectively, significantly attenuated the EA induced inhibition for at least 30 min. The K-receptor antagonist (nor-BNI) was less effective and was shorter acting. Like EA, microinjection of mu- and delta opioid agonists, DAGO and DADLE, respectively, into the rVLM significantly decreased the pressor responses. In contrast, the kappa-opioid agonist, U50,488, failed to alter the BK-induced pressor response. We conclude that a significant portion of inhibition of the gallbladder pressor response by EA is related to activation of mu- and delta-opioid receptors in the rVLM. The endogenous neurotransmitters for mu- and delta-opioid receptors, beta-endorphins and enkephalins, in the rVLM, therefore appear to play a role in the EA-related modulation of cardiovascular reflex responses. Conversely, dynorphin is less likely to be involved in this response. PMID- 11474646 TI - Sympathetic neurons of the cat stellate ganglion in postnatal ontogenesis: morphometric analysis. AB - Basic morphometric parameters (the maximal diameter, cross-sectional area), the distribution density of neurons were determined in the zones of the emergence of the basic nerves and in the center of the stellate ganglion (SG) in newborn, 10-, 20-day-old as well as 1- and 2-month-old kittens. Most of the investigated neurons in all animals were oval in their profile. In parallel to the increase of the average diameters, the number of small neurons decreased and the percentage of large neurons increased in postnatal ontogenesis. In all kittens, neurons with larger average size were located in the cranial pole of the ganglion. In 10-day old kittens and older animals, the average size of neurons was more in the left SG in comparison to the right one. Only in 1-month-old kittens the density of location of neurons in the right ganglion exceeded such parameters of neurons located at the left side. The number of neurons in the SG was not constant in postnatal ontogenesis and reduced from newborn to 20-day-old animals. PMID- 11474647 TI - Cardiorespiratory reflexes in a working heart-brainstem preparation of the house musk shrew, Suncus murinus. AB - In this study, we adapted the working heart-brainstem preparation (WHBP) from rodents to the Insectivore, Suncus murinus. Suncus WHBPs had a baseline heart rate of 333 +/- 8 beats min(-1), a perfusion pressure of 69 +/- 2 mm Hg and a respiratory cycle length of 6.5 +/- 0.7 s. Administration of atropine produced an increase in heart rate of 26 +/- 9 beats min(-1) indicative of the presence of cardiac vagal tone. Activation of baroreceptors produced pressure-dependent reflex falls in heart rate and reduced respiratory cycle length. The baroreceptor reflex sensitivity in Suncus WHBP was a decrease in heart rate of 8.1 +/- 1.4 beats min(-1) mm Hg(-1). Activation of peripheral chemoreceptors with aortic injections of sodium cyanide (0.1-12.5 microg) produced a dose-dependent reflex fall in heart rate and reduced respiratory cycle length. The reflex falls in heart rate evoked by baroreceptor and peripheral chemoreceptor stimulation were both atropine-sensitive. We conclude that viable WHBP can be prepared from Suncus and that Suncus WHBP is a novel non-rodent model in which to study brainstem mediated reflexes. PMID- 11474648 TI - A functional study of uncrossed and crossed pulmonary afferent fibres in the cervical vagus nerves of the cat. AB - The functional distribution of uncrossed and crossed pulmonary afferent fibres in the cervical vagus nerves has been studied in the anaesthetized cat using acute and chronic unilateral pneumonectomized preparations. The heart and lungs were sympathectomized routinely. The vagal afferent pathways of three pulmonary reflexes were investigated: the Hering-Breuer respiratory reflex, the lung inflation cardio-accelerator reflex, and the pulmonary chemoreflex. Inflation of the remaining lung caused temporary inhibition of inspiration. It also resulted in acceleration of the heart, but only when the background cardiac vagal tone was augmented. These respiratory and cardiac responses were abolished in most animals by ipsilateral cervical vagotomy; however, in some, a small response persisted and this was abolished by contralateral vagotomy. Stimulation of pulmonary C fibre endings with right atrial injections of phenylbiguanide caused a reduction in respiration, bradycardia and systemic hypotension, responses which occurred with a latency of 2.9 +/- 0.15 s. They were mostly abolished by ipsilateral cervical vagotomy, but reduced responses persisted in a few animals. The residual responses were abolished by contralateral cervical vagotomy and by selective denervation of the lung. These results indicate that most afferent fibres subserving the three pulmonary reflexes studied run in the ipsilateral cervical vagus, representing the uncrossed pathway. Some afferent fibres, however, cross to the contralateral cervical vagus. Degenerative changes in cells of the contralateral nodose ganglion in chronic unilateral pneumonectomized animals support these findings. PMID- 11474649 TI - Distribution of cGMP in guinea pig autonomic ganglia after stimulation with sodium nitroprusside. AB - Nitric oxide (NO) is an intercellular messenger molecule in the nervous system and exerts its action in many regions by generating cyclic GMP (cGMP) via soluble guanylyl cyclase. In this study, on the male guinea pig, we have analyzed the localization of cGMP in some autonomic ganglia with immunohistochemistry after stimulation with sodium nitroprusside (SNP) as NO donor, and made correlations with the NO synthesizing enzyme NO synthase (NOS), tyrosine hydroxylase (TH) and some neuropeptides. The putative target neurons for NO were examined in the anterior pelvic ganglia (APGs), as well as some pre- and paravertebral sympathetic ganglia. The results show that cGMP-like immunoreactivity (LI) in the APG was in most cases observed in the TH-positive, NOS-negative neuron population after SNP stimulation, whereas the NOS-expressing cholinergic population mostly lacked detectable cGMP-LI. In the pre- and paravertebral ganglia, SNP stimulation increased cGMP levels to a much lesser extent than in the APGs. cGMP was also observed in blood vessels, in the ganglion capsule, and in some cases. possibly in satellite cells. We propose, as one alternative, that there is a functional, intraganglionic regulatory loop between the parasympathetic and sympathetic divisions of the APG, using the NO/cGMP pathway. PMID- 11474650 TI - Peripheral target contact regulates Ca2+ channels in the cell bodies of bullfrog sympathetic ganglion B-neurons. AB - Tyrosine-hydroxylase immunohistochemistry demonstrated that a single injection of 120 mg/kg 6-hydroxydopamine (6-OHDA) reversibly disconnected bullfrog sympathetic ganglia from their peripheral targets. This was correlated with a decrease in sympathetic outflow to the eyes and a reversible decrease in pupil diameter. 6 OHDA did not damage the cell bodies of ganglionic neurons. Calcium channel current in ganglionic B-neurons, (measured at -10 mV; holding potential -60 mnV; Ba2+ as charge carrier; IBa) was reduced. It reached a minimum of about 40% of control amplitude 7-14 days after 6-OHDA injection and recovered to 73% of control amplitude after 63 days. 6-OHDA induced loss and recovery of functional sympathetic innervation of peripheral target tissues, as determined by measurement of pupil diameter, occurred at a similar rate. Thus, pupil diameter attained mininum values 7-14 days after 6-OHDA treatment and recovered to 81% of control after 63 days. The properties of Ca2+ channels in sympathetic neurons are, therefore, determined by continuity of contact with peripheral target. 6 OHDA also decreased the peak amplitude and duration of the afterhyperpolarization (a.h.p) that follows the action potential (a.p.). The rate of recovery of a.h.p duration was more rapid than the rate of recovery of peak a.h.p. amplitude. This may reflect known differences in properties of two types of Ca2+-sensitive K currents. IC and IAHP, IC, which is responsible for the peak amplitude of the a.h.p has a low affinity for Ca2+, whereas IAHP, which determines a.h.p. duration, has higher Ca2+ affinity. PMID- 11474651 TI - Attenuation of the afferent limb of the baroreceptor reflex in streptozotocin induced diabetic rats. AB - Diabetic autonomic neuropathy is a common complication following prolonged diabetes. Alterations of cardiovascular reflexes contribute to the increased cardiovascular morbidity and mortality seen in diabetic patients. This study sought to better characterize these complications by investigating the afferent limb of the baroreceptor reflex in an experimental rat model of diabetes. Streptozotocin (STZ)-induced diabetic and euglycemic control rats were studied at 8- and 16-week time points after initiation of the experiment. Activation of the afferent limb of the baroreceptor reflex was assessed by measuring the numbers of c-Fos-immunoreactive (ir) neurons in the CNS site of termination of the baroreceptor afferent neurons, the nucleus of the solitary tract (NTS). Initial experiments established that baseline cardiovascular parameters and NTS expression of c-Fos-ir neurons were not different between diabetic and control rats at either time point. Phenylephrine (PE)-induced activation of baroreceptors resulted in a significant elevation in the numbers of c-Fos-ir neurons in the NTS of control rats. Although diabetic rats showed similar pressor responses to PE, the activation of c-Fos-ir neurons in the NTS of diabetic rats was significantly attenuated. At both 8 and 16 weeks, STZ-induced diabetic rats had significantly fewer c-Fos-ir neurons in the commissural NTS and in the caudal subpostrernal NTS when compared to the non-diabetic control animals receiving PE. These data suggest that STZ-induced diabetes, for a period of 8 and 16 weeks, results in reduced activity in the afferent baroreceptor input to the NTS, and are consistent with diabetes-induced damage to baroreceptor afferent nerves. PMID- 11474652 TI - Orthostatic intolerance in Behcet's disease. AB - Neurological manifestations are known to occur in patients with Behcet's Disease (BD) and significantly affect the clinical course of the disease. Nevertheless, the prevalence, pattern and severity of autonomic impairment in such patients have yet to be defined. In this paper, we presented a BD patient with orthostatic tachycardia. Non-invasive standardized autonomic function tests revealed no evidence of autonomic impairment, but profound orthostatic tachycardia accompanied by abnormal catecholamine increase was observed upon standing. The diagnosis of Orthostatic Intolerance (OI) was made and initial symptomatic therapy was started. The contribution of an immunological damage to components of neural pathways in the pathogenesis of the autonomic involvement can be assumed. PMID- 11474653 TI - Pseudoxanthoma elasticum: evidence for the existence of a pseudogene highly homologous to the ABCC6 gene. PMID- 11474654 TI - Hereditary and somatic DNA mismatch repair gene mutations in sporadic endometrial carcinoma. PMID- 11474655 TI - Absence of learning difficulties in a hyperactive boy with a terminal Xp deletion encompassing the MRX49 locus. PMID- 11474656 TI - Identification of a new TWIST mutation (7p21) with variable eyelid manifestations supports locus homogeneity of BPES at 3q22. PMID- 11474657 TI - Association between the defective Pro369Ser mutation and in vivo intrahepatic 1 antitrypsin accumulation. PMID- 11474658 TI - The androgen receptor and DXS15-134 markers show a high rate of discordance for germline X chromosome inactivation. PMID- 11474659 TI - Evaluation of the ELOVL4 gene in families with retinitis pigmentosa linked to the RP25 locus. PMID- 11474660 TI - Parity, oral contraceptives, and the risk of ovarian cancer among carriers and noncarriers of a BRCA1 or BRCA2 mutation. AB - BACKGROUND: Multiparity and the use of oral contraceptives reduce the risk of ovarian cancer, but their effects on this risk in women with a BRCA1 or BRCA2 mutation are unclear. METHODS: We conducted a population-based case-control study of ovarian cancer among Jewish women in Israel. Women were tested for the two founder mutations in BRCA1 and the one founder mutation in BRCA2 that are known to be common among Jews. We estimated the effects of parity and oral contraceptive use on the risk of ovarian cancer in carriers and noncarriers in separate analyses that included all control women, who did not have ovarian cancer. RESULTS: Of 751 controls who underwent mutation analysis, 13 (1.7 percent) had a BRCA1 or BRCA2 mutation, whereas 244 of 840 women with ovarian cancer (29.0 percent) had a BRCA1 or BRCA2 mutation. Overall, each additional birth and each additional year of use of oral contraceptives were found to lower the risk of ovarian cancer, as expected. Additional births were protective in separate analyses of carriers and noncarriers, but oral-contraceptive use appeared to reduce the risk only in noncarriers; among carriers, the reduction in the odds of ovarian cancer was 12 percent per birth (95 percent confidence interval, 2.3 to 21 percent) and 0.2 percent per year of oral-contraceptive use ( 4.9 to 5.0 percent). CONCLUSIONS: The risk of ovarian cancer among carriers of a BRCA1 or BRCA2 mutation decreases with each birth but not with increased duration of use of oral contraceptives. These data suggest that it is premature to use oral contraceptives for the chemoprevention of ovarian cancer in carriers of such mutations. PMID- 11474661 TI - Efficacy of the anti-CD22 recombinant immunotoxin BL22 in chemotherapy-resistant hairy-cell leukemia. AB - BACKGROUND: Hairy-cell leukemia that is resistant to treatment with purine analogues, including cladribine, has a poor prognosis. We tested the safety and efficacy of an immunotoxin directed against a surface antigen that is strongly expressed by leukemic hairy cells. METHODS: RFB4(dsFv)-PE38 (BL22), a recombinant immunotoxin containing an anti-CD22 variable domain (Fv) fused to truncated pseudomonas exotoxin, was administered in a dose-escalation trial by intravenous infusion every other day for a total of three doses. RESULTS: Of 16 patients who were resistant to cladribine, 11 had a complete remission and 2 had a partial remission with BL22. The three patients who did not have a response received low doses of BL22 or had preexisting toxin-neutralizing antibodies. Of the 11 patients in complete remission, 2 had minimal residual disease in the bone marrow or blood. During a median follow-up of 16 months (range, 10 to 23), 3 of the 11 patients who had a complete response relapsed and were retreated; all of these patients had a second complete remission. In 2 of the 16 patients, a serious but completely reversible hemolytic-uremic syndrome developed during the second cycle of treatment with BL22. Common toxic effects included transient hypoalbuminemia and elevated aminotransferase levels. CONCLUSIONS: BL22 can induce complete remissions in patients with hairy-cell leukemia that is resistant to treatment with purine analogues. PMID- 11474663 TI - Glanders in a military research microbiologist. PMID- 11474662 TI - Treatment of chronic plaque psoriasis by selective targeting of memory effector T lymphocytes. AB - BACKGROUND: Psoriatic plaques are characterized by infiltration with CD4+ CD45RO+ and CD8+ CD45RO+ memory effector T lymphocytes. The recombinant protein alefacept binds to CD2 on memory effector T lymphocytes, inhibiting their activation. METHODS: In a multicenter, randomized, placebo-controlled, double-blind study, we evaluated alefacept as a treatment for psoriasis. Two hundred twenty-nine patients with chronic psoriasis received intravenous alefacept (0.025, 0.075, or 0.150 mg per kilogram of body weight) or placebo weekly for 12 weeks, with follow up for 12 additional weeks. Before treatment, the median scores on the psoriasis area-and-severity index were between 14 and 20 in all groups (0 denotes no psoriasis and 72 the most severe disease possible). RESULTS: Alefacept was well tolerated and nonimmunogenic. The mean reduction in the score on the psoriasis area-and-severity index two weeks after treatment was greater in the alefacept groups (38, 53, and 53 percent in the groups receiving 0.025, 0.075, and 0.150 mg per kilogram, respectively) than in the placebo group (21 percent, P<0.001). Twelve weeks after treatment, 28 patients who had received alefacept alone were clear or almost clear of psoriasis. Three patients in the placebo group were clear or almost clear; all three had received additional systemic therapy for psoriasis. Alefacept reduced peripheral-blood memory effector T-lymphocyte (CD45RO+) counts, and the reduction in the number of memory-effector T lymphocytes was correlated with the improvement in psoriasis. CONCLUSIONS: Treatment with alefacept for 12 weeks is associated with improvement in chronic plaque psoriasis; some patients have a sustained clinical response after the cessation of treatment. Alefacept selectively targets CD45RO+ memory effector T lymphocytes, suggesting that they have a role in the pathogenesis of psoriasis. PMID- 11474664 TI - Images in clinical medicine. Bilateral ovarian dermoid cysts. PMID- 11474665 TI - Clinical practice. Subclinical hypothyroidism. PMID- 11474666 TI - Treatment of endometriosis. PMID- 11474667 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 23-2001. An 18-month-old girl with persistent diarrhea and malnutrition. PMID- 11474668 TI - A magic bullet for cancer--how near and how far? PMID- 11474669 TI - New treatments for psoriasis. PMID- 11474670 TI - Ready or not--preparedness for bioterrorism. PMID- 11474671 TI - A synthetic pentasaccharide for the prevention of deep-vein thrombosis. PMID- 11474672 TI - A synthetic pentasaccharide for the prevention of deep-vein thrombosis. PMID- 11474673 TI - Low-molecular-weight heparin in patients with deep-vein thrombosis. PMID- 11474674 TI - Low-molecular-weight heparin in patients with deep-vein thrombosis. PMID- 11474675 TI - Biventricular pacing in patients with heart failure. PMID- 11474676 TI - Underuse of coronary revascularization procedures. PMID- 11474677 TI - Underuse of coronary revascularization procedures. PMID- 11474678 TI - Underuse of coronary revascularization procedures. PMID- 11474679 TI - Underuse of coronary revascularization procedures. PMID- 11474680 TI - Cutaneous squamous-cell carcinoma. PMID- 11474681 TI - Cutaneous squamous-cell carcinoma. PMID- 11474682 TI - Cutaneous squamous-cell carcinoma. PMID- 11474683 TI - Sterile cerebrospinal fluid ascites and chronic peritonitis. PMID- 11474684 TI - What's happening to Britain's National Health Service? PMID- 11474685 TI - Calcium channel blockers provide significant benefit in hypertension, says a WHO ISH analysis. PMID- 11474686 TI - Significant hypertrophy regression with indapamide sustained release 1.5 mg. PMID- 11474687 TI - Will ACE inhibitor therapy reduce secondary stroke? PMID- 11474688 TI - Benefits of combination therapy for achieving goal blood pressure in high CV risk patients. PMID- 11474689 TI - Aggressive LDL-cholesterol lowering with atorvastatin results in regression of atherosclerosis. PMID- 11474690 TI - Weight loss reduces risk of developing cardiovascular disease (CVD)--need for effective weight loss strategies grows as prevalence of obesity increases. PMID- 11474691 TI - Primary results of COPERNICUS, a pivotal landmark study (Carvedilol Prospective Randomised Cumulative Survival Trial). PMID- 11474692 TI - Lisinopril shows regression of myocardial fibrosis in patients with hypertensive heart disease. PMID- 11474693 TI - TACTICS-TIMI 18 shows positive results of invasive approach. PMID- 11474695 TI - Intravascular Ultrasound (IVUS) study exposes diffuse nature of atherosclerotic disease. PMID- 11474694 TI - Indapamide plus perindopril--simpler treatment, better compliance. PMID- 11474696 TI - Carvedilol outclasses the other beta-blockers. PMID- 11474697 TI - Carvedilol saves lives--new data from landmark trials prove survival benefits in heart failure and post myocardial infarction. PMID- 11474698 TI - LIFE study--still-blinded results show promise. PMID- 11474699 TI - Treatment of the lipoprotein complex increases health benefits. PMID- 11474700 TI - Unique vulnerability of diabetic patients to hypertensive injury. PMID- 11474701 TI - Irbesartan achieves consistent and effective use over 1 year. PMID- 11474702 TI - Perindopril lowers blood pressure without reducing cerebral blood flow in stroke patients with carotid disease. PMID- 11474703 TI - Study at up to 700 sites will build on landmark HOPE trial. PMID- 11474704 TI - Hepatitis B vaccine: risks and benefits of universal neonatal vaccination. AB - Global eradication of hepatitis B, which has infected over 2000 million people worldwide, is an achievable goal. Hepatitis B vaccine is effective and safe, and is recommended in Australia as a four-dose childhood schedule commencing with a neonatal dose. A neonatal dose has a greater impact on carriage, the main reservoir of transmission, due to the inverse relationship of age and risk of chronic carriage. Universal vaccination is clearly cost-effective in countries of high hepatitis B endemicity but less so in countries of low endemicity. Other factors affecting the perceived benefits of universal vaccination in low-risk countries include the use of the preservative thiomersal in hepatitis B vaccines, and case reports of multiple sclerosis (MS) and unexplained fever in recipients. Careful epidemiological studies have failed to confirm any risk of MS or fever with the hepatitis B vaccine, which is now thiomersal-free. Other arguments against universal vaccination include 'unnecessary' vaccination of low-risk neonates. However, selective vaccination programmes targeting at-risk neonates are often poorly implemented and do not protect against horizontal transmission in early childhood. Universal vaccination, which is safe and effective, is the only practical means of achieving global eradication of hepatitis B. PMID- 11474705 TI - Environmental triggers of type 1 diabetes. AB - High risk HLA class II alleles account for 40% of the genetic susceptibility to type 1 diabetes in Caucasians, but the majority of the genetically predisposed do not develop the disease. This supports some environmental modification of the autoimmune destruction of beta cells that precedes type 1 diabetes. Identical twin studies and geographical variation in incidence also argue for a critical role of environmental factors. Attention has been directed to the possible harmful effect of cow's milk protein (or protective effect of breast-feeding) and enteric infections in early life. Natural history studies that follow children at increased risk of type 1 diabetes provide the best opportunity to study environmental triggers. The Australian Baby Diab Study has followed approximately 500 babies from birth who have a first-degree relative with type 1 diabetes. No prospective association between duration of breast-feeding or introduction of cow's milk and the development of islet autoimmunity was found. The same Australian cohort demonstrated a relationship between rotavirus infection and the first appearance or increase in islet antibodies. Enteroviral infection is seen more frequently in prediabetic children and prior to the onset of islet autoimmunity in Finnish cohorts. Environmental factors may interact. Breast milk protects against enteric infections; enteric infections in turn could increase immunity to dietary antigens by increasing intestinal permeability. It is also possible that an alteration in gut mucosal immune function in genetically susceptible individuals underlies any effect of dietary or viral proteins on the development of islet autoimmunity in early life. PMID- 11474706 TI - Low cyclosporin-A level due to Saint-John's-wort in heart transplant patients. PMID- 11474707 TI - Managing a bilateral pneumothorax in lung transplantation using single chest-tube drainage. PMID- 11474708 TI - Combined anti-coagulation protocol for the MicroMed DeBakey VAD: a proposal. PMID- 11474709 TI - In vitro activities of mitomycin C against growing and hypoxic dormant tubercle bacilli. PMID- 11474710 TI - [Words and action of medical instruction: surgical lectures of Antonio Scarpa]. AB - Antonio Scarpa (1752-1832) was chairman of anatomy and surgery at the university of Pavia for thirty years since 1783. The study of the manuscripts containing his surgical lectures, collected by his pupils, is the basis for the knowledge of the method of teaching in its twofold dimension of the theoretical matter and of the practical demonstration. Starting from the analysis of one of these manuscripts, entitled "Lezioni di operazioni chirurgiche del Cav. Prof. Antonio Scarpa" the value of these collections of notes is discussed and their significance as a textbook for the students is emphasized. The survey of every single lecture demonstrates that the theoretical exposition immediately preceded the operative procedure on the corpse. Finally the qualities expected by Scarpa in the teachers and the students for a satisfactory education in surgery are considered. Fulfilling his mastership Scarpa struggled to convey to the pupils the method learned when he himself was a student. He was therefore considered the head of the anatomy and surgery school in Pavia. PMID- 11474711 TI - [Psychiatry and psychiatric care. On the relationship of science and society around 1900]. AB - Psychiatrists undertaking historical research, poets praising advances in the natural sciences, sociologists speaking the language of medicine and biology: these are only some examples of how various approaches started to flow together around 1900. This convergence was attributable on the one hand to a fear of social disintegration and the consequences of modernization, and on the other hand to a widespread belief in scientific progress. The nineteenth century concept of individual integrity and a right to cultural education was now replaced by collective approaches emphasizing the role of society as a whole (the "masses") and heredity. During the next few decades eugenic approaches to the treatment of "asocial elements" and minorities predominated. Rather than deriving, as claimed, from scientific principles, these approaches were often based on cultural argumentation. The trend was more toward a culturalization of the scientific and medical discourse than toward a biologization of thinking with regard to society. A good many areas in this connection have hardly been researched yet. Interdisciplinary studies would be required to elucidate the relationships between medically defined "health" and socially defined "normality", the intertwinement of disciplines, and the implementation of scientific concepts into social and political models. PMID- 11474714 TI - [Signs of discovery. (Re-)construction of infertility in the age of reproduction medicine]. AB - Even though infertility seems to be an old phenomenon, it will be argued that it was discovered in a new way once it became necessary to legitimize the technological possibilities of artificial fertilization at the end of the 1970s. With the birth of the first test-tube baby in 1978 the public became excited about the new possibilities of medical procreation. Reactions were extremely divided because extracorporeal fertilization seemed to undo the difference between natural organisms and technologically created human beings. Today reproductive medicine is an accepted field of gynecology and an increasing number of couples make use of it (even in managing their fertility). This development was the result of the successful integration of reproductive techniques into the discourse of infertility which is by no means self-evident. From the perspective of how societies communicate about reproduction it becomes clear that the dividing line between fertility and infertility is disappearing. PMID- 11474716 TI - [History of medicine and pharmacology. Aspects of the history of interdisciplinary relations]. AB - Both boundaries and communication exist between medical disciplines. Chairs for the combination of pharmacology and medical history were created in the 19th century. Some medical historians have dealt with pharmacology and its history (Heischkel, Sigerist, Temkin, Ackerknecht, Lesky, Mettler). In turn, individual pharmacologists have contributed to the history of medicine (Kobert, Leake, Binz, Hans Fischer), the last two particularly to its institutionalization. PMID- 11474717 TI - [First International Congress of Psychiatry and Neurology 1907]. AB - The first International Congress of Psychiatry, Psychology and Neurology took place in Amsterdam in 1907 with the participation of a remarkable number of the best-known specialists of the time. The author persuades a study of the arrangements made prior to the Congress and the reports published subsequently. The difference of opinion between the supporters of Freudian psychoanalysis and those of traditional neuropsychiatry that existed at that time was expressed only half-heartedly. Parallels with the great, modern congresses are drawn. PMID- 11474718 TI - Who attains social status? Effects of personality and physical attractiveness in social groups. AB - One of the most important goals and outcomes of social life is to attain status in the groups to which we belong. Such face-to-face status is defined by the amount of respect, influence, and prominence each member enjoys in the eyes of the others. Three studies investigated personological determinants of status in social groups (fraternity, sorority, and dormitory), relating the Big Five personality traits and physical attractiveness to peer ratings of status. High Extraversion substantially predicted elevated status for both sexes. High Neuroticism, incompatible with male gender norms, predicted lower status in men. None of the other Big Five traits predicted status. These effects were independent of attractiveness, which predicted higher status only in men. Contrary to previous claims, women's status ordering was just as stable as men's but emerged later. Discussion focuses on personological pathways to attaining status and on potential mediators. PMID- 11474719 TI - Changes in women's assertiveness in response to status and roles: a cross temporal meta-analysis, 1931-1993. AB - Across two meta-analyses, American women's assertiveness rose and fell with their social status from 1931 to 1993. College women and high school girls' self reports on assertiveness and dominance scales increased from 1931 to 1945, decreased from 1946 to 1967, and increased from 1968 to 1993, explaining about 14% of the variance in the trait. Women's scores have increased enough that many recent samples show no sex differences in assertiveness. Correlations with social indicators (e.g., women's educational attainment, women's median age at first marriage) confirm that women's assertiveness varies with their status and roles. Social change is thus internalized in the form of a personality trait. Men's scores do not demonstrate a significant birth cohort effect overall. The results suggest that the changing sociocultural environment for women affected their personalities, most likely beginning in childhood. PMID- 11474720 TI - Fear, anger, and risk. AB - Drawing on an appraisal-tendency framework (J. S. Lerner & D. Keltner, 2000), the authors predicted and found that fear and anger have opposite effects on risk perception. Whereas fearful people expressed pessimistic risk estimates and risk averse choices, angry people expressed optimistic risk estimates and risk-seeking choices. These opposing patterns emerged for naturally occurring and experimentally induced fear and anger. Moreover, estimates of angry people more closely resembled those of happy people than those of fearful people. Consistent with predictions, appraisal tendencies accounted for these effects: Appraisals of certainty and control moderated and (in the case of control) mediated the emotion effects. As a complement to studies that link affective valence to judgment outcomes, the present studies highlight multiple benefits of studying specific emotions. PMID- 11474721 TI - Do response time limitations counteract the effect of faking on personality inventory validity? AB - Different models of lying on personality scales make discrepant predictions on the association between faking and item response time. The current research investigated response time restriction as a method for reducing the influence of faking on personality scale validity. In 3 assessment simulations involving 540 university undergraduates responding to 2 common, psychometrically strong personality inventories, no evidence emerged to indicate that limiting respondents' answering time can attenuate the effects of faking on validity. Results were interpreted as failing to support a simple model of personality test item response dissimulation that predicts that lying takes time. Findings were consistent with models implying that lying involves primitive cognitive processing or that lying may be associated with complex processing that includes both primitive responding and cognitive overrides. PMID- 11474722 TI - Do people aggress to improve their mood? Catharsis beliefs, affect regulation opportunity, and aggressive responding. AB - Do people aggress to make themselves feel better? We adapted a procedure used by G. K. Manucia, D. J. Baumann, and R. B. Cialdini (1984), in which some participants are given a bogus mood-freezing pill that makes affect regulation efforts ineffective. In Study 1, people who had been induced to believe in the value of catharsis and venting anger responded more aggressively than did control participants to insulting criticism, but this aggression was eliminated by the mood-freezing pill. Study 2 showed similar results among people with high anger out (i.e., expressing and venting anger) tendencies. Studies 3 and 4 provided questionnaire data consistent with these interpretations, and Study 5 replicated the findings of Studies 1 and 2 using measures more directly concerned with affect regulation. Taken together, these results suggest that many people may engage in aggression to regulate (improve) their own affective states. PMID- 11474723 TI - Moral credentials and the expression of prejudice. AB - Three experiments supported the hypothesis that people are more willing to express attitudes that could be viewed as prejudiced when their past behavior has established their credentials as nonprejudiced persons. In Study 1, participants given the opportunity to disagree with blatantly sexist statements were later more willing to favor a man for a stereotypically male job. In Study 2, participants who first had the opportunity to select a member of a stereotyped group (a woman or an African American) for a category-neutral job were more likely to reject a member of that group for a job stereotypically suited for majority members. In Study 3, participants who had established credentials as nonprejudiced persons revealed a greater willingness to express a politically incorrect opinion even when the audience was unaware of their credentials. The general conditions under which people feel licensed to act on illicit motives are discussed. PMID- 11474724 TI - Do others judge us as harshly as we think? Overestimating the impact of our failures, shortcomings, and mishaps. AB - When people suffer an embarrassing blunder, social mishap, or public failure, they often feel that their image has been severely tarnished in the eyes of others. Four studies demonstrate that these fears are commonly exaggerated. Actors who imagined committing one of several social blunders (Study 1), who experienced a public intellectual failure (Studies 2 and 3), or who were described in an embarrassing way (Study 4) anticipated being judged more harshly by others than they actually were. These exaggerated fears were produced, in part, by the actors' tendency to be inordinately focused on their misfortunes and by their resulting failure to consider the wider range of situational factors that tend to moderate onlookers' impressions. Discussion focuses on additional mechanisms that may contribute to overly pessimistic expectations as well as the role of such expectations in producing unnecessary social anxiety. PMID- 11474725 TI - The norm of self-interest and its effects on social action. AB - Four studies investigated whether people feel inhibited from engaging in social action incongruent with their apparent self-interest. Participants in Study 1 predicted that they would be evaluated negatively were they to take action on behalf of a cause in which they had no stake or in which they had a stake but held stake-incongruent attitudes. Participants in Study 2 reported both surprise and anger when a target person took action on behalf of a cause in which he or she had no stake or in which he or she held stake-incongruent attitudes. In Study 3, individuals felt more comfortable engaging in social action and expected others to respond more favorably toward their actions if the issue was described as more relevant to their own sex than to the opposite sex. In Study 4, the authors found that providing nonvested individuals with psychological standing rendered them as likely as vested individuals to undertake social action. The authors discuss the implications of these results for the relationship between vested interest, social action, and attitude-behavior consistency. PMID- 11474726 TI - Trust and communicated attributions in close relationships. AB - The attributional statements intimate partners communicate to one another were examined as a function of trust. In discussions by 35 married couples, 850 attributions and corresponding events were coded on dimensions of valence, globality, and locus. Results of regression and contingency analyses indicate that attributional statements expressed in high-trust relationships emphasized positive aspects of the relationship. Medium-trust couples actively engaged issues but focused more on negative events and explanations. Low-trust couples expressed more specific, less affectively extreme attributional statements that minimized the potential for increased conflict. Results could not be accounted for by relationship satisfaction. These findings also highlight the importance of focusing on features of the events for which attributions are expressed. PMID- 11474727 TI - Attitude alignment in close relationships. AB - On the basis of principles of balance theory and interdependence theory, this research examined a phenomenon termed attitude alignment, or the tendency of interacting partners to modify their attitudes in such a manner as to achieve attitudinal congruence. The results of three experiments generally were consistent with the proposed model. First, tendencies toward attitude alignment were greater to the extent that attitudinal discrepancies were salient. Second, alignment tendencies were greater to the extent that an issue was central to the partner; there was also evidence that the degree to which an issue was peripheral to the self affected alignment processes (e.g., for changes in centrality of issue, with regard to persuasion methods). Third, degree of alignment tended to be greater in dating-partner interactions than in stranger interactions and tended to be greater among couples with high adjustment than among those with low adjustment. PMID- 11474728 TI - Mind-set and close relationships: when bias leads to (In)accurate predictions. AB - The authors investigated whether mind-set influences the accuracy of relationship predictions. Because people are more biased in their information processing when thinking about implementing an important goal, relationship predictions made in an implemental mind-set were expected to be less accurate than those made in a more impartial deliberative mind-set. In Study 1, open-ended thoughts of students about to leave for university were coded for mind-set. In Study 2, mind-set about a major life goal was assessed using a self-report measure. In Study 3, mind-set was experimentally manipulated. Overall, mind-set interacted with forecasts to predict relationship survival. Forecasts were more accurate in a deliberative mind-set than in an implemental mind-set. This effect was more pronounced for long-term than for short-term relationship survival. Finally, deliberatives were not pessimistic; implementals were unduly optimistic. PMID- 11474729 TI - Attachment theory and intergroup bias: evidence that priming the secure base schema attenuates negative reactions to out-groups. AB - Five studies examined the effects of priming the secure base schema on intergroup bias. In addition, Studies 1-2 examined the effects of dispositional attachment style, Studies 2-5 examined a mood interpretation, Study 3 examined the mediating role of threat appraisal, and Studies 4-5 examined the effects of secure base priming while inducing a threat to self-esteem or cultural worldview. Secure base priming led to less negative evaluative reactions toward out-groups than positive affect and neutral control conditions. In addition, whereas the effects of secure base priming did not depend on attachment style and were not explained by mood induction, they were mediated by threat appraisal and occurred even when self esteem or cultural worldview was threatened. The discussion emphasizes the relevance of attachment theory for understanding intergroup attitudes. PMID- 11474730 TI - The acute toxicity and primary irritancy of glutaraldehyde solutions. AB - Glutaraldehyde (GA, CAS Number 110-30-8), an aliphatic dialdehyde, has a wide range of industrial, scientific, and medical applications. It is available in aqueous solutions, whose concentrations vary up to 50% (w/w) and from which there is a potential during use for skin and eye contact and exposure to the vapor. The acute toxicity and primary irritancy of a wide range of GA concentrations were investigated to determine the differential hazards for such solutions. The acute peroral toxicity in the rat, expressed as ml of solution dosed, was moderate for solutions of 5% and above (LD50 range 0.88-3.25 ml/kg) and generally varied little for solutions up to 50%. Solutions less than 5% GA were of slight toxicity (LD50 range 3.34-12.30 ml/kg for 1 and 2% solutions). When lethality was expressed as absolute amount of GA dosed (mg GA/kg), there was a reciprocal relationship between the concentration of GA solution dosed and LD50. This was confirmed in the mouse, which is more susceptible than the rat to acute peroral toxicity. The acute percutaneous toxicity of GA solutions to rabbits (24 h occlusion) was moderate (LD50 range 1.59-2.71 ml/kg) for 46 and 50% solutions, and slight for 25% GA solutions (8.80-16.00 ml/kg). At 15% and less, 16.0 ml/kg was not lethal. Exposures (4-8 h) of rats to saturated vapor atmospheres of GA generated dynamically or statistically at ambient temperature (17-25 C) produced only transient peripheral sensory irritant effects to the eyes and respiratory tract. In contrast, vapor atmosphere generated dynamically at elevated temperature (60 or 65 C) produce severe effects, including mortality (4-h LC50 range 23.5-44.3 ppm). Histopathology in rats that died included exposure concentration-related acute inflammation and necrosis in the nasal mucosa, larynx, trachea, and bronchi. Standard primary skin irritation tests in the rabbit indicated severe skin irritation and necrosis at 45 and 50% GA; necrosis occurred with 1 and 4 h contact at 50% and at 4 h with 45%. Inflammation was moderate at 25%, slight to moderate with 5 and 10% GA, minor at 2%, and threshold at 1%. Standard primary eye irritation tests showed 45% GA to produce severe conjunctival and corneal injury, which was persistent. At 2% GA corneal injury was mild, and at 5% marked. The lowest concentration producing corneal injury was 1.0%, and the no-effects concentration was 0.5%. The threshold for conjunctival effects was 0.2%, and the no-effects concentration 0.1%. At 1% GA, conjunctival hyperemia and chemosis were moderate to marked, and became more severe with higher GA concentrations. The results suggest potential acute handling hazards with various concentrations of GA solutions and indicate industrial hygiene considerations. PMID- 11474731 TI - A case of jellyfish sting. AB - Jellyfish sting may result in a wide range of symptoms from common erythematous urticarial eruptions to the rare box-jelly induced acute respiratory failure. In Taiwan, with the increasing frequency of international travel, cases of jellyfish sting to foreigners are on the rise. We report a case of jellyfish sting with the rare presentation of painless contact dermatitis. A 38-y-o man accidentally stepped on a sea urchin with his right foot during scuba diving in a beach in Thailand. Traditional therapy with vinegar was applied on the lesion. However, when he returned to Taiwan, erythematous patches on the left thigh with linear radiations to the leg were discovered. The skin lesions had bizzare shapes and showed progressive change. No pain or numbness was noticed. Jellyfish stingwas suspected, topical medications were applied, and the patient recovered without complication. Jellyfish stings usually result in a painful erythematous eruption. In this case, though the lesion involved a large surface, there was no pain. Delayed diagnosis of jellyfish sting was due to the atypical presentation and the physician's unfamiliarity to the Thai jellyfish sting. Awareness to the wide spectrum of jellyfish sting symptoms should be promoted. PMID- 11474732 TI - Bovine atypical interstitial pneumonia associated with the ingestion of damaged sweet potatoes (Ipomoea batatas) in Northeastern Brazil. AB - Atypical interstitial pneumonia in cattle associated with the ingestion of damaged sweet potatoes is reported in the State of Paraiba, Northeastern Brazil. The sweet potatoes were severely damaged by Myzus tersicae, and had an obvious fungal infection. Eighteen milking cows, 1 bull and 1 steer were fed approximately 400 kg of the sweet potatoes. Six days after consumption 13 cows were affected with labored abdominal breathing, extended and lowered head, coughing, expiratory grunt, salivation and protruded tongue. Six animals died and the others recovered in 4-7 d. At necropsy lungs were distended and did not collapse when the thorax was opened. Bullous emphysema and gelatinous exudates were observed in the interlobular, peribronchial and subpleural tissues. Microscopically, the lungs had severe edema and emphysema, congestion, and alveolar epithelial cell hyperplasia. The sweet potatoes were cultured but Fusarium spp was not isolated, probably because a zigomycete fungus covered the plates in 48 h of incubation. PMID- 11474733 TI - Late confirmation of acute methyl bromide poisoning using S-methylcysteine adduct testing. AB - Methyl bromide poisoning is difficult to confirm because routine laboratory testing has not been reliable. Measurable levels of the parent agent are rapidly reduced, probably as a result of direct tissue chemical reaction. Serum bromide levels have been used as an indirect measure of exposure and/or toxicity but are inconsistent. Recently special testing has shown that protein adducts formed after exposure to methyl bromide may be a better measure of significant exposure. The S-methylcysteine adduct was used to confirm acute methyl bromide toxicity 10 weeks after an exposure. Additionally, genotyping of the glutathione-S transferase TI enzyme in erthyrocytes from this case characterized a predisposition to the neurotoxic effects of methyl bromide. PMID- 11474734 TI - Fatal poisoning by butachlor and chlornitrofen ingested from a bottle marked as nitrofen. AB - Nitrofen has been banned in Taiwan since January 1, 1983 due to its tetrogenicity. A 78-y-o female consumed about 500 ml of herbicide, labeled as nitrofen, in a suicide attempt. Consciousness disturbance occurred immediately. After 9 h, bloody-tarry stool was noted. Hematemesis occurred 19 h later. Hepatotoxicity and nephrotoxicity also developed. Twitching of head and both upper extremities lasting 20-30 sec and up to 5 min developed on day 3. Sometimes an upward gaze with the face turned to the right or a convulsion of the right upper extremity was observed. EEG showed alpha coma, generalized slow spike-and wave complexes, and isolated generalized sharp waves mainly in the left occipital area. On day 7, endotracheal intubation was performed due to apnea; she expired on day 9. The pesticide was found to contain butachlor and chlornitrofen. The Agricultural Committee prohibited the marketing of chlornitrofen due to its generation of tumors in animals. The combination of butachlor and chlornitrofen can result in consciousness disturbance, leucocytosis, gastrointestinal hemorrhage, rhabdomyolysis, hypocalcemia, hypoalbuminemia, elevated amylase, nephrotoxicity, hepatotoxicity, seizures and death. The cytotoxicity of butachlor may be related to the patient's death. Active components of pesticides should be checked if the clinical course of a poisoned patient is unusual to allow appropriate interventions. PMID- 11474735 TI - Myoclonus secondary to the concurrent use of trazodone and fluoxetine. AB - A 39-y-o male with a history of human immunodeficiency virus infection and depression was admitted for diagnosis and treatment of tuberculosis and pneumocystis carinii pneumonia infections. Prior to admission, he was on 50 mg trazodone every evening for 2 mo for depression. He was admitted with a 2-w history of fever chills and fatigue and on admission had hand tremors which disappeared at rest. Four days post-admission the trazodone dose was increased to 100 mg and 20 mg fluoxetine was initiated. He became increasingly anxious and his hand tremor worsened 3 d after initiation of the regimen. To rule out drug induced tremor, both trazodone and fluoxetine were discontinued and symptoms resolved in 7 d. Clinicians should be aware of the potential for excessive seratonergic activities secondary to trazodone + fluoxetine interactions causing a worsening myoclonus adverse event. PMID- 11474736 TI - Thapsia garganica L: a poisonous plant of North Africa. AB - Poisoning of livestock by ingestion of Thapsia garganica L, common plant in many countries of North Africa, is described. PMID- 11474737 TI - Persistently elevated acetaminophen concentrations for two days after an initial four-hour non-toxic concentration. AB - The decision to treat patients who overdose with n-acetylcysteine is routinely made with a single APAP concentration drawn 4 or more hours post-ingestion. However, in cases where there are co-ingestants that may delay gastric emptying, there have been recommendations for additional concentrations to determine peak APAP concentrations. This report is of a case of acetaminophen overdose involving narcotic co-ingestants with persistenty elevated acetaminophen concentrations for 2 d, suggesting delayed gastric emptying and/or bezoar formation. A second striking feature of this case was the persistently elevated acetaminophen concentrations without evidence of liver injury despite antidotal therapy not being employed. PMID- 11474738 TI - Analysis of research studying the effects of polychlorinated biphenyls and related chemicals on neurobehavioral development in children. AB - During the past 25 y numerous studies have been performed in infants and children to determine whether environmental exposure to polychlorinated biphenyls (PCBs), dibenzo-p-dioxins, dibenzofurans (PCDFs) and dichlorodiphenyl trichloroethane is causally associated with neurobehavioral effects. In addition, 1 episode of poisoning with PCDFs occurred in Taiwan in 1978. This poisoning outbreak is referred to as Yu-Cheng, oil disease, since the PCDFs and PCBs had accidentally contaminated cooking oil. In the aggregate the studies reviewed here show no conclusive evidence that environmental exposure to these chemicals affect the neurobehavioral development of infants and children. PMID- 11474739 TI - Dimethyl sulfoxide: recent pharmacological and toxicological research. AB - A survey of the literature published in the past 2 decades on the basic pharmacology, therapeutic uses and toxicity of dimethyl sulfoxide (DMSO) is presented. A salient pharmacological action of DMSO is its ability to scavenge oxygen-free radicals implicated in xenobiotic-induced tissue damages when given before, during or several hours after the tissue insult. More trials with DMSO in diseases and conditions caused by oxygen-free radicals are warranted. PMID- 11474740 TI - Immunomodulative effects of aflatoxins and selenium on human natural killer cells. AB - The possible immunomodulative effects of aflatoxin B1 and selenium (as sodium selenite) at concentrations lower than 5.10(-11) M or 0.05ppb and 10(-5) M, respectively on natural killer (NK) cells from healthy volunteers were evaluated by a 51Cr release natural killer assay. Aflatoxin B1 and selenium separately had an immunosuppressive effect on NK cell activity. When they were studied in combination, they caused a statistically significant increase of the cytotoxic activity of NK cells in the presence of the lowest concentrations of aflatoxin B1 (0.005ppb) and of sodium selenite (10(-8)M). PMID- 11474741 TI - Let go of Latin! AB - Health care practitioners have long used Latin abbreviations. Using these shorthand terms does not consistently promote patient safety. although in some instances they offer a time-saving convenience. This article illustrates hazards associated with the common Latin abbreviation "Qd" by showing copies of actual medication orders. Other dangerous abbreviations are also discussed. The use of plain English is suggested as the prescribing practice most consistent with professional values. PMID- 11474742 TI - Treatment of palmar hyperhidrosis with botulinum toxin. AB - Excessive sweating of the palms, axillae, and soles can be managed with intradermal injections of botulinum toxin as an alternative to more aggressive surgical therapies such as sympathectomy and less effective techniques including topical antiperspirants. The dosage and injection techniques can be optimized to provide several months of freedom from this troubling disorder. PMID- 11474743 TI - Complications and adverse reactions with the use of botulinum toxin. AB - Botulinum toxins are the causative agents of the severe food-borne illness botulism. With lethal doses approximating 10(-9) g/kg body weight, these neurotoxins represent some of the most toxic naturally occurring substances. Regardless, botulinum toxin is considered a safe therapy for inappropriate muscle spasms with adverse effects being typically self-limited. This article deals with some of the complications that have occurred with these treatments. The greatest concern with the use of BOTOX is probably the formation of blocking antibodies leading to nonresponse of subsequent treatment. Prevalence of resistance is less than 5%. Most complications associated with its aesthetic use are few and anecdotal. Nevertheless, the common problems and pitfalls associated with aesthetic treatment of the various areas of the face and neck with botulinum toxin are discussed. Also included are recommendations as to how to avoid these very undesirable, yet common, problems. PMID- 11474744 TI - Other noncosmetic uses of BOTOX. AB - Botulinum toxin A has a wide variety of clinical applications, which are related by blockade of acetylcholine and often are related to abnormal muscle contractures. These applications include ocular disorders, disorders of the upper aerodigestive tract, dystonia and hemifacial spasm, cosmetic, gastrointestinal disorders, genitourinary disorders, management of pain, and use in autonomic nervous system disorders. Many of these diseases will be discussed with regard to their treatment with botulinum toxin compared to conventional treatments. Advantages and disadvantages of botulinum toxin use are delineated. General guidelines for adult and pediatric dosing will also be discussed. PMID- 11474745 TI - Botulinum toxin type B: an overview of its biochemistry and preclinical pharmacology. AB - Produced by Clostridium botulinum, botulinum toxins are high molecular weight protein complexes consisting of the neurotoxin and additional nontoxic proteins that function to protect the toxin molecule. The neurotoxin acts to inhibit the release of acetylcholine at the neuromuscular junction, causing muscle paralysis. Purified toxin complexes have found a niche in the treatment of clinical disorders involving muscle hyperactivity. The different serotypes are structurally and functionally similar; however, specific differences in neuronal acceptor binding sites, intracellular enzymatic sites, and species sensitivities suggest that each serotype is its own unique pharmacologic entity. Recently, botulinum toxin type B has been developed as a liquid formulation to avoid the lyophilization (vacuum-drying) and reconstitution processes associated with decreasing the potency and stability of current type A toxin preparations. Biochemical tests were conducted to evaluate the quality of toxin in this formulation. In 3 consecutive manufacturing lots, the botulinum toxin type B complex was found to be highly purified, intact, uniform, and consistent from lot to lot. Also, it showed long-term stability at refrigerator and room temperatures (2 to 25 degrees C). Electrophysiologic studies in cynomolgus monkeys showed that botulinum toxin type B is effective in paralyzing injected muscle groups, with minimal spread to relatively distant noninjected muscles. PMID- 11474746 TI - Botulinum toxins. Introduction. PMID- 11474747 TI - Botulinum toxin type A: history and current cosmetic use in the upper face. AB - This article reviews the cosmetic use of botulinum toxin in upper face from both the historic and clinical viewpoints. The published literature and our current experience are outlined. Botulinum toxin type A in the upper face has become an extremely poplular cosmetic procedure and is outstandingly safe. PMID- 11474748 TI - BOTOX use in the mid and lower face and neck. AB - BOTOX is the universally accepted gold standard treatment for upper facial rejuvenation. BOTOX application in the mid and lower face and neck is a more advanced treatment requiring a detailed knowledge of underlying facial muscular anatomy and function as well as the patient's aesthetic desires and an ability to interpret the effects of BOTOX chemodenervation on their function. PMID- 11474749 TI - Botulinum toxin type A (BOTOX) for treatment of migraine. AB - An open-label study and 2 double-blind, placebo-controlled studies have provided supporting evidence of botulinum toxin type A (BTX-A) as an effective, well tolerated treatment for migraine. Observed durations of benefit were consistent with known properties of BTX-A. Findings suggest that response may vary by features of preinjection headaches, such as migraine frequency. The precise mechanism by which BTX-A provides pain relief is hypothesized to be related not only to acetylcholine inhibition but also to a blocking action on the parasympathetic nervous system. Additional studies that control factors likely to be related to response may lead to better understanding of the BTX-A effect on migraine and an optimal treatment protocol. PMID- 11474750 TI - Subjective well-being among the old-old: the role of health, personality and social support. AB - The purpose of this study was to devise and test a path model that explains how background variables, physical capacity, and psychosocial resources (locus of control and social network supportiveness) affect three well-being outcome measures--self-rated health, mental health, and life satisfaction--among the old old in Israel. A sample of European-born persons aged seventy-five and over, drawn randomly from the population registry in the greater Tel Aviv area, was administered a structured questionnaire in personal interviews (N=194). The findings confirm that the relationship between background variables, physical capacity and well-being is differentially mediated by the psychosocial resources, with different variable combinations predicting each of the respective facets of well-being. Moreover, the relative strength of the relationships between locus of control and social network supportiveness and well-being indicate that among the old-old, personality factors may be more consequential than social resources for one's well-being. PMID- 11474751 TI - Everyday interpersonal conflicts of middle-aged women: an examination of strategies and their contextual correlates. AB - Everyday problem solving within the interpersonal domain has been identified as propelling cognitive development in adulthood, and may be particularly relevant to women's development. Yet few studies of everyday problem solving in adulthood focus on interpersonal problems, on women, or on the middle adult years. We examined strategies middle-aged women used for real-life interpersonal problems, and contextual variables associated with strategy use. In contrast to research with younger and older women, in which lower-level strategies were favored, strategy use here was fairly evenly split between higher and lower levels. Also in contrast to other research with younger and older women, participants here favored lower-level strategies that were directive, rather than submissive. Thematic analyses showed a high degree of felt emotional distress to be associated with lower-level strategy use, and attempts at controlling emotions and overcoming fundamental attribution errors in judgment to be related to higher level strategy use. PMID- 11474752 TI - Support from others and efficacy of volunteer ombudsmen in long-term care facilities. AB - The work of the ombudsman has been described as the most difficult in the field of aging. Hypotheses about support from ten sources, relationships with peers, and efficacy were tested using questionnaire data from 607 volunteer ombudsmen in nursing facilities. Support from facility administrators and residents contributed to efficacy whereas help from other volunteers had a negative effect. Committed peers, however, facilitated the perception of a job well done. Efficacious ombudsmen believed their needs were taken into account suggesting the importance of caring for volunteers who in turn care for others. PMID- 11474753 TI - Identity processing styles and the need for self-esteem in middle-aged and older adults. AB - This study was a test of the relationship between self-esteem and the identity processing styles of identity assimilation (i.e., maintaining consistent views of the self), accommodation (i.e., changing the self ), and a balance between consistency seeking and identity change. A community sample of 242 older adults ranging in age from forty to ninety-five (M = 63.31) completed measures of identity processing and self-esteem. Previous research has demonstrated that identity assimilation increases with age in order to maintain self-esteem in the domain of physical and cognitive functioning; this is referred to as the identity assimilation effect (IAE). Based on this research, a similar result was expected in the domain of personality. Although identity assimilation and balance predicted increases in self-esteem, and identity accommodation predicted decreases in self-esteem, as predicted, no interaction effects were observed. The results of this study suggest the IAE may be domain specific to physical and cognitive functioning. PMID- 11474754 TI - Marital relationship in late adulthood: synchronous versus asynchronous couples. AB - The article presents a study on marital relations in late adulthood, conducted among a sample of 469 Israeli couples, who were divided into four groups based on occupational status and spousal timing of retirement. Two of the groups were categorized as synchronous: pre-retired couples (both partners are close to retirement); and retired couples (both partners are retired), and two of the groups were categorized as asynchronous: employed husband/retired wife and retired husband wife/employed husband. Differences between the groups were examined in three areas: Division of tasks in the home; marital power relations; and quality of marriage. Regarding division of tasks in the home, feminine tasks were usually more egalitarian among retired couples and retired husbands/employed wives than among pre-retired couples and employed husbands/retired wives. In addition, synchronous-retired couples were found to be more egalitarian than synchronous-pre-retired and asynchronous employed husbands/retired wives with regard to general tasks, while masculine tasks were usually carried out by husbands in all four groups. With respect to quality of marriage, the pre-retired couples expressed more marital complaints than did the retired couples, while no differences were found between either of the asynchronous groups. Nonetheless, marital power relations were generally egalitarian in all four groups. PMID- 11474755 TI - Motor vehicle crash involvement and moving violations: convergence of self-report and archival data. AB - In the crash involvement literature, it is generally assumed that archival and other "objective" criterion data are superior to self-reports of crash involvement. Using 394 participants (mean age = 36.23 years), the present study assessed the convergence of archival and self-report measures of motor vehicle crash involvement and moving violations. We also sought to determine whether predictor/criterion relationships would vary as a function of criterion type (i.e., archival vs. self-report), and if a combination of both criteria would result in better prediction than would either by itself. The degree of agreement between the two criterion sources was low, with participants self-reporting more crashes and tickets than were found in their state records. Different predictor/criterion relationships were also found for the two criterion types; stronger effects were obtained for self-report data. Combining the two criteria did not result in relationships stronger than those obtained for self-reports alone. Our findings suggest that self-report data are not inherently inferior to archival data and, furthermore, that the two sources of data cannot be used interchangeably. Actual or potential applications include choosing the appropriate criterion to use, which, as the finding of this study reveals, may depend on the purpose of the investigation. PMID- 11474756 TI - Eye activity correlates of workload during a visuospatial memory task. AB - Changes in six measures of eye activity were assessed as a function of task workload in a target identification memory task. Eleven participants completed four 2-hr blocks of a mock anti-air-warfare task, in which they were required to examine and remember target classifications (friend/enemy) for subsequent prosecution (fire upon/allow to pass), while targets moved steadily toward two centrally located ship icons. Target density served as the task workload variable; between one and nine targets were simultaneously present on the display. For each participant, moving estimates of blink frequency and duration, fixation frequency and dwell time, saccadic extent, and mean pupil diameter, integrated over periods of 10 to 20 s, demonstrated systematic changes as a function of target density. Nonlinear regression analyses found blink frequency, fixation frequency, and pupil diameter to be the most predictive variables relating eye activity to target density. Participant-specific artificial neural network models, developed through training on two or three sessions and subsequently tested on a different session from the same participant, correlated well with actual target density levels (mean R = 0.66). Results indicate that moving mean estimation and artificial neural network techniques enable information from multiple eye measures to be combined to produce reliable near real-time indicators of workload in some visuospatial tasks. Potential applications include the monitoring of visual activity of system opetators for indications of visual workload and scanning efficiency. PMID- 11474757 TI - Auditory distraction and short-term memory: phenomena and practical implications. AB - Irrelevant sound tends to break through selective attention and impair cognitive performance. This observation has been brought under systematic scrutiny by laboratory studies measuring interference with memory performance during exposure to irrelevant sound. These studies established that the degree of interference depends on the properties of the irrelevant sound as well as those of the cognitive task. The way in which this interference increases or diminishes as characteristics of the sound and of the cognitive task are changed reveals key functional characteristics of auditory distraction. A number of important practical implications that arise from these studies are discussed, including the finding that relatively quiet background sound will have a marked effect on efficiency in performing cognitive tasks. PMID- 11474758 TI - Effects of hearing protectors on auditory localization in azimuth and elevation. AB - An experiment was conducted to determine the effects of two types of hearing protectors on auditory localization performance. Six listeners localized a 750-ms broadband noise from loudspeakers ranging in azimuth from -180 degrees to +180 degrees and in elevation from -75 degrees to +90 degrees. Independent variables included the type of hearing protector and the elevation of the source. Dependent measures included azimuth error, elevation error, and the percentage of trials resulting in a front-back confusion. Performance on each of the dependent measures was found to be mediated by one or more of the independent variables. Actual or potential applications include the generation of improved design guidelines for hearing protectors and workplace alarms. PMID- 11474759 TI - Effects of image scale and system time delay on simulator sickness within head coupled virtual environments. AB - Novel patterns of visual-vestibular intersensory stimulation often result in symptoms of simulator sickness, raising health and safety concerns regarding virtual environment exposure. Two experiments investigated the effect of conflicting visual-vestibular cues on subjective reports of simulator sickness during and after a 50-min exposure to a head-coupled virtual interface. Virtual image scale factors (0.5. 1.0, 2.0 magnification, generated by varying geometric field of view angle) were investigated in Experiment 1, and additional system time delays (125, 250 ms) were investigated in Experiment 2. Simulator sickness metrics included spoken self-reports during exposure and simulator sickness questionnaires (pre-exposure, immediate postexposure, and 20 min postexposure). Head yaw angular position data were also recorded. Reports of simulator sickness symptoms were significantly greater in the minification (0.5) and magnification (2.0) image scale factor conditions than in the neutral condition (1.0). Simulator sickness did not vary with changes in time delay, however. Furthermore, a comparison across experiments suggests no appreciable increase in simulator sickness with increasing time delays above the nominal value (48 ms). Head angular position data exhibited certain systematic variations across conditions. Actual or potential applications of this research include virtual environment training, simulation, and entertainment systems. PMID- 11474760 TI - Spatial representations of virtual mazes: the role of visual fidelity and individual differences. AB - Twenty-four people learned three versions of a room-sized maze: a wire-frame desktop virtual environment (VE), a normal surface-rendered desktop VE, and a real-world maze. Differences among the mental representations formed from each environment were measured with pointing and distance estimation tasks in a real world version of each maze. People were more accurate at pointing after having learned the real and wire-frame VE maze than the surface-rendered VE maze; however, this effect was small compared with the effect of individual differences. Differences in gender, spatial ability, and prior computer experience were significantly related to the ability to acquire spatial information from the desktop VE. There was a high correlation between spatial knowledge when it was measured in the VE and spatial knowledge measured in the real world. Actual or potential applications include the design of effective VE training systems. PMID- 11474761 TI - Minimum and comfortable driving headways: reality versus perception. AB - A field study was conducted to evaluate drivers' actual headways in car-following situations, their relationship to the drivers' brake reaction times, and their relationship to the drivers' ability to estimate those headways using different metrics. Drivers were asked to maintain "minimum safe distance" and "comfortable, normal distance with no intention to pass" behind the car ahead. The lead car speeds varied from 50 to 100 km/hr. The results showed that under both sets of instructions, drivers adjusted their distance headways in relation to speed, maintaining constant time headways. A significant portion of the drivers maintained time headways that are considered unsafe in relation to drivers' reaction times. There was no significant relationship between the minimal headways maintained by the drivers and their brake reaction times under conditions of maximum attention and preparedness to apply brakes. Accuracy of spoken estimates of headways varied widely among the three measures used to report perceived headway; meters and car lengths yielded much lower estimates (and ones closer to the actual headways) than did seconds. The results have implications for headway perception, driving safety, driver education, and smart cruise-control design. PMID- 11474762 TI - Peripheral visual feedback: a powerful means of supporting effective attention allocation in event-driven, data-rich environments. AB - Breakdowns in human-automation coordination in data-rich, event-driven domains such as aviation can be explained in part by a mismatch between the high degree of autonomy yet low observability of modern technology. To some extent, the latter is the result of an increasing reliance in feedback design on foveal vision--an approach that fails to support pilots in tracking system-induced changes and events in parallel with performing concurrent flight-related tasks. One possible solution to the problem is the distribution of tasks and information across sensory modalities and processing channels. A simulator study is presented that compared the effectiveness of current foveal feedback and two implementations of peripheral visual feedback for keeping pilots informed about uncommanded changes in the status of an automated cockpit system. Both peripheral visual displays resulted in higher detection rates and faster response times, without interfering with the performance of concurrent visual tasks any more than does currently available automation feedback. Potential applications include improved display designs that support effective attention allocation in a variety of complex dynamic environments, such as aviation, process control, and medicine. PMID- 11474763 TI - Effects of computer monitor viewing angle and related factors on strain, performance, and preference outcomes. AB - A model of visual and musculoskeletal strain associated with computer monitor placement was developed. The main premise of which is that monitor placement decisions must take into consideration development of both visual and musculoskeletal strains. Certain factors in the model that were thought to affect one or both types of strain. or that were considered important to rule out for effect, were tested in a lab setting. These factors were viewing angle (eye level, midlevel, low level), monitor size (14 in., 19 in.), keyboard familiarity (touch typist, nontouch typist), and task (reading, mousing, typing). Outcomes included indicators of visual and musculoskeletal strain, preference, and performance. Muscle activity was generally greater for the low viewing angle, for the standard monitor (14 in.), and for non-touch typists. Participants preferred the midlevel placement. Task performance was slightly diminished with eye-level placement. Results are interpreted in relation to the model and to several hypotheses that were formed to focus the inquiry. Actual or potential applications of this research include monitor placement decisions in the design or modification of computer workstations. PMID- 11474764 TI - Visual context and the control of movements through video display. AB - The aim of this experiment was to test the influence of target context on adaptation to scale perturbations introduced by a video display. Participants performed pointing movements without direct vision of their moving hand, although they could see their movements on a video display. Their perceived movements could be reduced, enlarged. or displayed at their actual size. Three target contexts were compared: dark surround, illuminated frame, and familiar object. Movements were executed with or without vision of hand displacement. Results showed that target context enhanced an allocentric coding of the movement, which improved movement execution. However, the effect of target context changed whether or not the displacement of the hand was available. Overall, the results suggest that target context allowed the extraction of dynamic information about movements, which is used to program and control movements. This suggests that target context could be used efficiently to improve spatial accuracy and speed in teleoperation learning. Potential applications include the reduction of difficulties encountered during teleoperation learning through the introduction of visual context. PMID- 11474765 TI - Anesthesia alarms in context: an observational study. AB - This paper surveys current work on the design of alarms for anesthesia environments and notes some of the problems arising from the need to interpret alarms in context. Anesthetists' responses to audible alarms in the operating room were observed across four types of surgical procedure (laparoscopic, arthroscopic, cardiac, and intracranial) and across three phases of a procedure (induction, maintenance, and emergence). Alarms were classified as (a) requiring a corrective response, (b) being the intended result of a decision, (c) being ignored as a nuisance alarm, or (d) functioning as a reminder. Results revealed strong effects of the type of procedure and phase of procedure on the number and rate of audible alarms. Some alarms were relatively confined to specific phases; others were seen across phases, and responses differed according to phase. These results were interpreted in light of their significance for the development of effective alarm systems. Actual or potential applications of this research include the design of alarm systems that are more informative and more sensitive to operative context than are current systems. PMID- 11474766 TI - Does stress training generalize to novel settings? AB - Many high-stress task environments are complex and dynamic, and it is often difficult during training to anticipate the exact conditions that may be encountered in these settings. We conducted an empirical study to examine whether the positive effects of stress training that addressed one specific type of stressor and task would remain when trainees performed under a novel stressor or performed a novel task. Participants performed a laboratory task under stress conditions. Measures of task performance and self-reported stress were obtained at three performance trials: (a) prior to stress training, (b) after a stress training intervention targeted to that specific task environment, and (c) under novel stressor/task conditions. Results indicated that the beneficial effects of stress training were retained when participants performed under a novel stressor and performed a novel task. We discuss the implications of this study with regard to their application in the design of stress training and the transfer of learning to complex, dynamic task environments. PMID- 11474767 TI - Improving the treatment of epilepsy in the era of managed care. Introduction. PMID- 11474768 TI - Evaluation of the patient with epilepsy. AB - Accurate evaluation of the patient with epilepsy is the first step toward developing an effective treatment regimen. Many problems can occur when patients who have had seizures are not assessed properly. A seizure may be caused by conditions other than epilepsy. If epilepsy is the cause, knowing the type of seizure and identifying epilepsy syndromes as early as possible is vital. An incorrect diagnosis can leave the patient with uncontrolled disease, leading to debilitating morbidity that in most cases can be treated effectively. Patients who would be good candidates for surgical intervention benefit by being identified early to prevent long periods, perhaps years, with uncontrolled disease. Understanding the type of seizure or epilepsy syndrome also determines the type of antiepileptic drugs that should be selected. Early, accurate evaluation and diagnosis are 2 of the most important aspects of treatment. PMID- 11474769 TI - The role of new antiepileptic drugs. AB - For many years, epileptologists had few choices for treating seizures. Within the past 20 years several "new generation" antiepileptic drugs (AEDs) were introduced. The most recent additions include oxcarbazepine, levetiracetam, and zonisamide. New agents have been shown in clinical trials to offer similar efficacy compared with older, more established AEDs, but the new agents offer important improvements in safety. Although clinical trials to specifically measure the efficacy of the new AEDs in treating idiopathic generalized epilepsy are rare, the new agents have demonstrated efficacy in treating generalized tonic clonic convulsions. Data for treatment of Lennox-Gastaut syndrome indicate a clear effect with lamotrigine or topiramate and possibly some effect with zonisamide and levetiracetam. Studies of juvenile myoclonic epilepsy and absence seizures suggest that zonisamide, lamotrigine, topiramate, and levetiracetam may be effective. Each of the new AEDs is effective in controlling partial seizures. These agents may also be appropriate choices for newly diagnosed patients or those whose conditions are refractory to treatment. In clinical trials, patients who are refractory to treatment are often given escalated doses to gain effect, but higher doses also result in more adverse events and higher withdrawal rates. Generally, the higher the dose, the greater the odds of withdrawal, with the exception of levetiracetam, which is not associated with increased withdrawal rates at high doses. Newly diagnosed patients are likely to be controlled with the first therapy given to them. It is therefore important to select an agent with the best safety, efficacy, and tolerability profile possible. PMID- 11474770 TI - Pharmacokinetics of the new antiepileptic drugs. AB - A physician's choice of an antiepileptic drug (AED) usually depends upon the patient's seizure type. But the pharmacokinetic characteristics of AEDs can further help determine the best drug for a particular patient. These characteristics, including absorption, elimination pathway, and potential for drug interactions, are of critical importance for patients who take other medication for comorbid conditions and for patients with impaired renal or hepatic function. The ideal AED would have a rapid, linear, consistent absorption rate with complete clearance, low plasma protein binding, and rapid central nervous system penetration. It would be eliminated predominantly by the kidneys. The new AEDs do not prompt the same concerns about interactions because they have much better pharmacokinetic profiles than the older drugs and as a result require less monitoring for potential interactions. PMID- 11474771 TI - Treatment of epilepsy in 3 specialized populations. AB - When discussing the treatment of epilepsy, targeted populations need to be defined. Three patient groups, children, the elderly, and women, are considered "special" because of metabolic and physical differences that require particular care during treatment of this disease. Treatment options vary significantly among these populations. Metabolic differences in very young and elderly patients require close attention by the prescribing physician. Rates of metabolism in children may be much faster than in nonelderly adults, requiring dosing adjustments to ensure enough medication is used to control seizures. Additional concerns with treating children include their increased sensitivity to toxic effects. Elderly patients may have slower metabolic rates because of decreased renal or hepatic function, and thus these patients can easily be overdosed as toxic drug levels build when clearance is reduced. Many elderly patients also may have concomitant illnesses that require other chronic medications. The potential for drug interactions is very high among this population. Women are considered a special population because of issues related to contraception, childbirth, and breast-feeding. Some antiepileptic medications are known to reduce the efficacy of oral contraceptives, and no medication has been proven safe for pregnant women. The pharmacokinetic profiles of many new generation antiepileptic medications may be advantageous to these specialized populations by creating fewer adverse effects, cleaner metabolism, and the reduced risk for drug interactions and teratogenicity. PMID- 11474772 TI - Managing healthcare resources appropriately. Roundtable discussion. PMID- 11474773 TI - CYP2C19 genotype and pharmacokinetics of three proton pump inhibitors in healthy subjects. AB - PURPOSE: To predict the CYP2C19 genotype-dependence in anti-Helicobacter pylori (H. pylori) therapy when lansoprazole or rabeprazole was used instead of omeprazole as a proton pump inhibitor (PPI). METHODS: A comparative pharmacokinetic study with each PPI was designed as an open, randomized, and crossover study of 18 Japanese healthy volunteers who were classified into the homozygous, heterozygous extensive metabolizer and the poor metabolizer based on the CYP2C19 genotype determined by PCR-RFLP method. Each subject received a single oral dose of 20 mg omeprazole, 30 mg lansoprazole, or 20 mg sodium rabeprazole, with at least 1 week washout period between treatments. Plasma concentrations of PPIs and their metabolites were monitored until 12 h after medication. RESULTS: Pharmacokinetic profiles of omeprazole and lansoprazole were well correlated with the CYP2C19 genotype. The heterozygous extensive metabolizer was slightly different from the homozygote, but there was no statistically significant difference. The CYP2C19 genotype dependence found for lansoprazole was not obvious compared with omeprazole. As for rabeprazole, the pharmacokinetic profile was independent of the CYP2C19 genotype. CONCLUSIONS: CYP2C19 genotype dependence will be found in the anti-H. pylori therapy even when lansoprazole is used as the PPI. PMID- 11474774 TI - Evaluation of the bioequivalence of highly-variable drugs and drug products. AB - PURPOSE: To establish procedures for the effective evaluation of bioequivalence (BE) for highly-variable drugs and drug products (HVD/P). METHODS: 2- and 4 period crossover BE studies with 24 subjects were simulated which generally assumed within-subject coefficients of variation of 40%. The relationship between the fraction of studies in which BE was accepted (the statistical power) and the ratio of geometric means (GMR) of the two formulations was evaluated for various methods of analysis. These included, primarily, scaled average BE (ABE), the corresponding approach of expanding BE limits (BEL), and, for comparison, unscaled ABE and scaled individual BE (IBE). RESULTS: Scaled ABE and expanding BEL showed very similar properties in both 2- and 4-period studies. They had steeper power curves than scaled IBE. Unscaled ABE had very low statistical power. The acceptance of BE by unscaled and scaled ABE and expanding BEL was almost independent of subject-by-formulation interaction and the ratio of within subject variations of the two formulations. By contrast, the conclusions reached by scaled IBE were strongly affected by these parameters. CONCLUSIONS: Scaled ABE and expanding BEL evaluate BE effectively for HVD/P in both 2- and 4-period investigations. However, additional, useful information can be obtained from 4 period studies. PMID- 11474775 TI - Novel direct curve comparison metrics for bioequivalence. AB - PURPOSE: The object of this work was to devise four new direct curve comparison (DCC) metrics and examine each metric's distribution properties and performance characteristics. METHODS: DCC metrics, Cmax, and AUCi were calculated from two bioequivalence studies of three sustained release carbamazepine formulations, where a range of profile similarity was observed. DCC metric values and their confidence intervals were compared to Cmax and AUCi. RESULTS: The DCC metrics rho, rhom, deltaa, and deltas, exhibited more favorable distributions than Cmax and AUCi ratios, which were frequently skewed. The DCC metrics performed differently than Cmax and AUCi ratios in profile comparisons due to the nature of the DCC metrics. Unlike Cmax and AUCi, the DCC metrics utilize all data points to directly compare entire profiles. Each DCC metric appears to measure "exposure" in a single assessment. Possible bioequivalence acceptance criteria are: p < or =1.40, rhom, < or =0.35, deltaa, < or =0.27, and deltas < or =0.102. CONCLUSIONS: These DCC metrics, particularly rhom, are promising bioequivalence metrics for "exposure." PMID- 11474776 TI - Human jejunal permeability of two polar drugs: cimetidine and ranitidine. AB - PURPOSE: To determine the human jejunal permeability of cimetidine and ranitidine using a regional jejunal perfusion approach, and to integrate such determinations with previous efforts to establish a baseline correlation between permeability and fraction dose absorbed in humans for soluble drugs. METHODS: A sterile multi channel perfusion tube, Loc-I-Gut, was inserted orally and positioned in the proximal region of the jejunum. A solution containing cimetidine or ranitidine and phenylalanine, propranolol, PEG 400, and PEG 4000 was perfused through a 10 cm jejunal segment in 6 and 8 subjects, respectively. RESULTS: The mean Peff (+/- se) of cimetidine and ranitidine averaged over both phases were 0.30 (0.045) and 0.27 (0.062) x 10(-4) cm/s, respectively, and the differences between the two were found to be statistically insignificant. The mean permeabilities for propranolol, phenylalanine, and PEG 400 averaged over both phases and studies were 3.88 (0.72), 3.36 (0.50), and 0.56 (0.08) x 10(-4) cm/s, respectively. The differences in permeability for a given marker were not significant between phases or between the two studies. CONCLUSIONS: The 10-fold lower permeabilities found for cimetidine and ranitidine in this study, compared to propranolol and phenylalanine, appear to be consistent with their less than complete absorption in humans. PMID- 11474777 TI - Anti-drug antibodies as drug carriers. I. For small molecules. AB - PURPOSE: To better understand the pharmacokinetics of drugs compounds that bind endogenous antibodies METHODS: Three groups of mice with differing anti fluorescein (FL) titers were established by empirically developed immunization protocols. These with two control groups were given intravenously [3H] ethanolamine conjugate of FL (FL-EA). The latter was synthesized using isothiocyanate chemistry. Radioactivity in the circulation, and occasionally in peritoneal ascites, was monitored for 7 days. A group of mice was immunized with eosin Y and given FL-EA. Conversely, eosin Y conjugate of radiolabeled EA (EY-EA) was given to mice immunized with FL. These two groups represented animals of low affinity to probe haptens. The affinity was assessed by a precipitation procedure, while titer was determined by a standard ELISA. Dose of FL-EA varied over a 100-fold. RESULTS: On average, the three immunized groups showed a 1:13:85 ratio of anti-FL titer, with remarkably consistent levels within each group. Elimination rates of FL-EA from the serum of very high-titer mice and high-titer mice were similar, however, were substantially lower than that found in low-titer mice. The latter was in turn lower than that found in non- or mock-immunized mice. Serum of mice immunized with FL showed approximately 200-fold lower affinity towards EY-EA than FL-EA. In these mice and in mice immunized with eosin Y and given FL-EA, the elimination of the probe haptens was again fast, reminiscent of low-titer mice. Mice of either low titer or low affinity showed more rapid redistribution of the conjugate between serum and peritoneal fluid. In a group of mice with comparable anti-FL titer, elimination from serum was independent of dose over a 100-fold difference. The bi-phasic concentration-time profile observed was accommodated by a physiologically meaningful pharmacokinetic model incorporating two compartments in which antibody binding can occur. CONCLUSIONS: Monovalent antigenic substance cannot trigger immune clearance. As such, endogenous antibodies that recognize the molecule can serve as a carrier to result in a substantial decrease in clearance. PMID- 11474778 TI - Antibodies as drug carriers. II. For proteins. AB - PURPOSE: To evaluate the potential use of antibodies as a carrier for monovalent protein haptens. METHODS: A single -SH functionality present in the human IgG light chain was fluoresceinated. This conjugate, FL-LC, was treated with pepsin to obtain FL conjugate of half light chain, FL-(LC)1/2, of MW 11 kDa. These two were radiolabeled using [3H]-propionic acid N-hydroxysuccinimide ester, and administered via tail vein to FL-immunized or mock-immunized mice. The blood radioactivity was measured over a 72-h period. Attempts were made to measure the affinity constant for the interaction between the conjugates and anti-FL antibodies by fluorescence quenching, surface plasmon resonance spectroscopy, and competitive ELISA. RESULTS: All of the three methods used produced supportive, if not conclusive, evidence of decreased binding affinity with increased conjugate size. Subsequent to tail-vein injection to FL-immunized mice, FL-LC showed approximately 4-fold smaller volume of distribution than mock-immunized mice: 0.041 +/- 0.005 vs. 0.16 +/- 0.02 mL/g. Corresponding values for FL-(LC)1/2, were significantly larger: 0.070 +/-0.013 and 0.30 +/- 0.02 mL/g, respectively. Compared with a small FL conjugate of ethanolamine, FL-EA, we studied earlier, the dose-normalized concentrations of the protein conjugates started at a higher level but declined more rapidly with time. In mock-immunized mice, the radioactivity disappeared very rapidly after administration, followed by an extremely slow decline with half-life close to 60 h. Evidence is provided to support that the radiolabel dissociated in the kidney, however, binding to anti FL antibodies greatly stabilized the conjugate. CONCLUSIONS: Based on an entropic principle alone the affinity of monovalent hapten-antibody interaction is expected to diminish with increase in hapten size. As such, the size of a hapten should be an important determinant of its pharmacokinetics in animals harboring antibodies that recognize the hapten. Relative to what was observed with small MW FL-EA, the protein conjugates showed substantially sustained circulation as a result of antibody binding, but this effect was diminished at later time points. Both affinity and pharmacokinetic data are consistent with the hypothesis of reduced affinity with increasing MW for monovalent hapten conjugates, but neither offered overwhelming proof. PMID- 11474779 TI - Simulation of the delivery of doxorubicin to hepatoma. AB - PURPOSE: To develop a two-dimensional simulation platform for the transport of doxorubicin to the hepatoma. To examine the temporal and spatial variation of doxorubicin concentration and its penetration into the tumor and the surrounding normal tissues. METHODS: Simulations are carried out with Fluent/UNS using the finite volume method to obtain the interstitial fluid pressure, velocity, and concentration profiles. RESULTS: Interstitial fluid pressure in the tumor and core reaches a steady state value in about 800 s, corresponding well with the assumed time scale for interstitial matrix fluid percolation (-1,000 s). There is a strong correlation between the drug concentration in the interstitial space of tumor and blood plasma for time >> 1 h. Concentration of doxorubicin is highest in the viable zone of the tumor at early times and in the necrotic core at later times, and lowest in the surrounding normal tissues. Diffusion is the dominant form of transport for doxorubicin. CONCLUSIONS: Varying the volume of solution injected, while keeping the dosage the same, does not cause significant changes in the amount and distribution of drug in the tumor. A higher vascular exchange area leads to higher concentrations of drug in the tumor. Lymphatic drainage in the tumor causes negligible reductions in the mean concentrations in all three different zones. Cellular metabolism and DNA binding kinetics decrease the mean concentrations of drug by about 15 to 40%, as compared to the baseline case. PMID- 11474780 TI - Systemic delivery of cetrorelix to rats by a new aerosol delivery system. AB - PURPOSE: To study the pulmonary absorption and tolerability of various formulations of the decapeptide cetrorelix acetate in rats by a new aerosol delivery system (ASTA-ADS) for intratracheal application. METHODS: Using the ASTA ADS, cetrorelix liquid formulations (aqueous solutions for ultrasonic nebulization) were firstly selected and subsequently delivered as nebulized aerosol to orotracheally cannulated rats. The pharmacologic effect (decrease of testosterone serum level) of four cetrorelix formulations was determined in rats by enzyme linked immunosorbant assay, and pharmacokinetic data were determined after measurement of cetrorelix serum level by radioimmunoassay. Histological examination of the lung was performed at the end of the experiments, and in a supplementary experiment the respiratory parameters (resistance and compliance) of rats were monitored by a validated pulmonary monitoring system during the aerosol application of the same formulations. RESULTS: After an exposure time of 5 min, the applied formulations reduced the testosterone concentration in serum to subnormal levels (< or =1 ng/ml) over a period of 24 h. Comparing the plasma concentration after intratracheal aerosolization with data of intravenous administration, the mean calculated bioavailabilities for the four formulations using the corrected dose (delivered--exhaled amount) were between 48.4 +/- 27.0% and 77.4 +/- 44.0%. The histologic examination of the lungs revealed different tolerability of the various tested formulations ranging from locally intolerable to well tolerated. The measurement of the lung function parameters did not reveal any compound or formulation related changes. CONCLUSIONS: Our studies show that cetrorelix can be effectively administered as aerosol and that intratracheal aerosolization via the ASTA-ADS provides results that are well comparable to other application routes, as demonstrated by statistical comparison of the newly obtained data with previous results from intratracheal instillation of cetrorelix solutions in rats. PMID- 11474781 TI - Host factors influencing the preferential localization of sterically stabilized liposomes in Klebsiella pneumoniae-infected rat lung tissue. AB - PURPOSE: To gain insight into the host factors influencing liposome localization at sites of bacterial infection. METHODS: In a unilateral Klebsiella pneumoniae pneumonia rat model, capillary permeability and number of circulating leukocytes was quantified and related to the degree of liposome target localization. RESULTS: Liposome localization was highest in the hemorrhagic zone of infection, a zone characterized by markedly increased capillary permeability and high bacterial numbers. Both liposome localization and capillary permeability correlated positively with severity of infection. Lung instillation of other inflammatory stimuli, such as lipopolysaccharide or 0.1 M HCl inducing increased capillary permeability, also promoted liposome localization. As liposomal target localization in leukopenic rats was similar to that in immunocompetent rats, contribution of circulating leukocytes seems limited. Intrapulmonary distribution of liposomes shows that leukocytes at the target site are involved in liposome uptake after extravasation. CONCLUSIONS: Increased capillary permeability plays a crucial role in liposome localization at the infected site, whereas contribution of leukocytes is limited. These results suggest inflammatory conditions that could benefit from liposomal drug delivery. The involvement of leukocytes in liposome uptake at the target site could be important information in the selection of appropriate drugs. PMID- 11474782 TI - Size-dependent bioadhesion of micro- and nanoparticulate carriers to the inflamed colonic mucosa. AB - PURPOSE: The size-dependent deposition of microparticles and nanoparticles after oral administration to rats using an experimental model colitis was examined. Local delivery of an entrapped drug could reduce side effects and would be a distinct improvement compared with existing colon delivery devices. METHODS: Ulcerative colitis was induced in Lewis rats with trinitrobenzenesulfonic acid. Fluorescent polystyrene particles with a size of 0.1, 1, or 10 microm were administered for 3 days. The animals then were sacrificed and their guts resected. Particle distribution in the colon was imaged by confocal laser scanning microscopy and quantified by fluorescence spectrophotometry. RESULTS: In the inflamed tissue, an increased adherence of particles was observed at the thicker mucus layer and in the ulcerated regions. A size dependency of the deposition was found, and an increased number of attached particles to the colon was determined compared with the control group. For 10-micorm particles, only fair deposition was observed (control group: 1.4 +/- 0.6%; colitis: 5.2 +/- 3.8% of administered particle mass). One-micrometer particles showed higher binding (control group: 2.0 +/- 0.8%; colitis: 9.1 +/- 4.2%). Highest binding was found for 0.1-microm particles (control group: 2.2 +/- 1.6%; colitis: 14.5 +/- 6.3%). The ratio of colitis/control deposition increased with smaller particle sizes. CONCLUSIONS: The use of submicron-sized carriers holds promise for the targeted delivery of drugs to the inflamed colonic mucosal areas in inflammatory bowel disease. PMID- 11474783 TI - Glucose-binding property of pegylated concanavalin A. AB - PURPOSE: Concanavalin A (Con A) has been used in the development of sol-gel phase reversible hydrogels for modulated insulin delivery. The usefulness of Con A has suffered from its poor aqueous solubility and stability. The goal of this study was to modify Con A with poly(ethylene glycol) (PEG) and examine the water solubility and stability of the PEGylated Con A. METHODS: Con A was PEGylated using monomethoxy poly(ethylene glycol) p-nitrophenol carbonates, and the extent of PEGylation was determined by the fluorescamine method. The stability of the PEGylated Con A was examined by measuring the time-dependent absorbance at 630 nm. The binding affinities of glucose and allyl glucose to native- and PEGylated Con A were measured by the equilibrium dialysis method. RESULTS: The total number of PEG molecules that can be grafted to Con A was 10. As the number of grafted PEG chains per each Con A was increased up to 5, the binding affinity of glucose was gradually increased and reached the maximum. The solubility and stability of PEGylated Con A were improved significantly over those of native Con A. The binding affinity of allyl glucose to Con A was not changed much by PEGylation. When the extent of PEGylation was excessive (i.e., the number of grafted PEG chains per each Con A was larger than 5), however, the binding affinities of both glucose and allyl glucose were decreased significantly. CONCLUSIONS: PEGylation of Con A resulted in improved aqueous solubility and stability of Con A. The binding affinity of glucose increased and reached the maximum when the extent of PEGylation was 50%. Advantages of PEGylated Con A over native Con A are improved aqueous solubility, enhanced long-term stability, and higher glucose sensitivity. PMID- 11474784 TI - HMG-CoA reductase inhibitors (statins) characterized as direct inhibitors of P glycoprotein. AB - PURPOSE: HMG-CoA reductase inhibitors (statins) are commonly prescribed for lipid lowering to treat hypercholesterolemia. Although they are well tolerated, their pharmacokinetic interactions with other drugs can lead to some adverse clinical consequences. The avenue of interaction has been asserted to be CYP3A4 because most (or all) known interactions are with CYP3A4 inhibitors, and statin oxidative metabolism is mediated by CYP3A4 as well as other CYP enzymes. However, these same drugs that exert a clinical pharmacokinetic effect on statin disposition are generally also P-gp substrates/inhibitors; hence, this transporter may be, or may contribute to, the mechanism of interaction. METHODS: This study shows directly, as well as quantifies, the inhibition of P-gp-mediated transport of a fluorescent marker substrate. RESULTS: Lovastatin and simvastatin are very potent and effective inhibitors of P-gp transport with IC50's of 26 and 9 microM, respectively, for the human enzyme. Atorvastatin is also an effective P-gp inhibitor, but at higher concentrations. Uniquely, pravastatin, whose functional groups render it an inferior inhibitor of P-gp in the whole cell, had no effect in this assay. This result is consistent with known clinical interactions. The effect of these statins on ATP consumption by P-gp was also assessed, and the Km results were congruent with the IC50 observations. CONCLUSIONS: Therefore, the clinical interactions of statins with other drugs may be due, in part or all, to inhibition of P-gp transport. PMID- 11474785 TI - PEPT2-mediated uptake of neuropeptides in rat choroid plexus. AB - PURPOSE: The peptide transporter PEPT2 was recently shown to be functionally active in rat choroid plexus, suggesting that it may play a role in neuropeptide homeostasis in the cerebrospinal fluid. This study, therefore, examined the role of PEPT2 in mediating neuropeptide uptake into choroid plexus. METHODS: Whole tissue rat choroid plexus uptake studies were performed on GlySar in the absence and presence of neuropeptides and on carnosine. RESULTS: The neuropeptides NAAG, CysGly, GlyGln, kyotorphin, and carnosine inhibited the uptake of radiolabeled GlySar at 1.0 mM concentrations. In contrast, TRH, [D-Arg2]-kyotorphin, glutathione, and homocarnosine did not inhibit GlySar uptake. Kyotorphin, an analgesic, was a competitive inhibitor of GlySar with a Ki of 8.0 microM. The direct uptake of carnosine was also shown to be mediated by PEPT2 in isolated choroid plexus (Km = 39.3 microM; Vmax = 73.9 pmol/mg/min). Radiolabeled carnosine uptake was inhibited by 1.0 mM concentrations of GlySar or carnosine but not homocarnosine, L-histidine, or beta-alanine. CONCLUSIONS: These findings indicate that PEPT2 mediates the uptake of a diverse group of neuropeptides in choroid plexus, and suggests a role for PEPT2 in the regulation of neuropeptides, peptide fragments, and peptidomimetics in cerebrospinal fluid. PMID- 11474786 TI - Excretion into gastrointestinal tract of irinotecan lactone and carboxylate forms and their pharmacodynamics in rodents. AB - PURPOSE: To investigate the excretion of irinotecan hydrochloride (CPT-11) and its active metabolite, SN-38, into the gastrointestinal lumen via the biliary and/or intestinal membrane route after dosing with lactone and carboxylate forms of CPT-11, and to evaluate the toxic and antitumor effects of the two forms. METHODS: The excretions of CPT-11 and SN-38 were investigated by the in situ perfusion technique using rats. The incidence of delayed diarrhea was evaluated after i.v. dosing (60 mg/kg) with CPT-11 lactone and carboxylate forms for 4 days. Antitumor activity and changes in body weight were investigated in mice with Meth A tumors. RESULTS: The excretion of CPT-11 into bile was greater in dosing with CPT-11 carboxylate than that with its lactone form, whereas the exsorption across intestinal membrane was greater in dosing with CPT-11 lactone than that with its carboxylate form. Dosing with CPT-11 lactone dose-dependently inhibited the increase in tumor weights in Meth A tumor mice, whereas the dosing with its carboxylate form reduced the antitumor effect. CONCLUSIONS: The decreased antitumor effect caused by dosing with the CPT-11 carboxylate form could be due to less accumulation in the tissue including tumor cells resulting from the rapid elimination of the form in the body. PMID- 11474787 TI - Intestinal absorption of octreotide using trimethyl chitosan chloride: studies in pigs. AB - PURPOSE: To investigate the enhancing effect of trimethyl chitosan chloride (TMC) on the enteral absorption of octreotide and to delineate the required doses of both TMC and peptide in vivo in juvenile pigs. METHODS: Six female pigs (body weight, 25 kg) were operated to induce a stoma at the beginning of their jejunum and to insert an in-dwelling fistula for intrajejunal (IJ) administration of the formulations. A silicone cannula was inserted at the jugular vein for blood sampling. One week after surgery the pigs received IJ octreotide solution administrations with or without TMC at pH 7.4 or chitosan HCl at pH 5.5. For determining bioavailability (F) values, the pigs also received an octreotide solution intravenously (IV). Blood samples were taken from the cannulated jugular vein and subsequently analyzed by radioimmunoassay. RESULTS: Intrajejunal administration of 10 mg octreotide without any polymer (control solution) resulted in F values of 1.7 +/- 1.1% (mean +/- SE). Chitosan HCl 1.5% (w/v) at pH 5.5 led to a 3-fold increase in F compared to the control (non-polymer containing) formulations. Co-administration of octreotide with 5 and 10% (w/v) TMC at pH 7.4 resulted in 7.7- and 14.5-fold increase of octreotide absorption, respectively (F of 13.9 +/- 1.3% and 24.8 +/- 1.8%). IJ administration of 5 mg octreotide solutions resulted in low F values of 0.5 +/- 0.6%, whereas co administration with 5% (w/v) TMC increased the intestinal octreotide bioavailability to 8.2 +/- 1.5%. CONCLUSIONS: Cationic polymers of the chitosan type are able to enhance the intestinal absorption of the peptide drug octreotide in pigs. In this respect, TMC at neutral pH values of 7.4 appears to be more potent than chitosan HCl at a weak acidic pH of 5.5. PMID- 11474788 TI - Gastrointestinal transit and mucoadhesion of colloidal suspensions of Lycopersicon esculentum L. and Lotus tetragonolobus lectin-PLA microsphere conjugates in rats. AB - PURPOSE: To investigate in vivo the fate and the behavior of lectin-particle conjugates after oral administration. METHODS: Two plant lectins were selected, namely Lycopersicon esculentum L. and Lotus tetragonolobus lectins, which have been reported to be specific for oligomers of N-acetyl-D-glucosamine and L fucose, respectively, and conjugated to small poly(lactide) microspheres. Their intestinal transit was investigated in detail using radiolabeled particles. The transport and the distribution of the particles along the intestine, as well as their interactions with the intestinal mucosa, were determined after oral administration in rat. RESULTS: The overall transit of the particles was shown to be strongly delayed when the microspheres were conjugated to the lectins, mainly due to the gastric retention of the particles. A significant fraction of the conjugates adhered to the gastric and intestinal mucosae. No significant differences were observed after a preliminary incubation of lectin-microsphere conjugates with specific sugars. CONCLUSION: Although specific interactions could not be excluded, especially in the stomach, it was likely that adhesion was predominantly due to nonspecific interactions. These results could be attributed both to unfavorable physicochemical characteristics of the conjugates and to premature adsorption of soluble mucin glycoproteins, preventing any further specific adhesion. PMID- 11474789 TI - Altered disposition and effect of lerisetron in rats with elevated alpha 1-acid glycoprotein levels. AB - PURPOSE: To examine the effect of changes in plasma alpha1-acid glycoprotein (AAG) levels on the pharmacokinetics (PK) and pharmacodynamics (PD) of lerisetron, a novel serotonin 5-HT3 receptor antagonist, in the rat. METHODS: After subcutaneous administration of turpentine oil, AAG was significantly elevated compared with controls. The PK of unchanged lerisetron (UL; high performance liquid chromatography with radioactivity monitoring) and total lerisetron (TL; unchanged + changed, scintillation counting) was characterized post intravenous (i.v.) 14C lerisetron (50 microg/kg) in control and turpentine oil pretreated rats. The PK (0-180 min) was described by a two-compartmental model. Protein binding of lerisetron in vitro was measured using an ultrafiltration technique. The effect of lerisetron (5 microg/kg, i.v.) over 180 min was measured in anesthetized rats (control and pretreated) with the Bezold Jarisch reflex (inhibition of bradycardia after 16 microg/kg serotonin i.v.) as the endpoint. PD parameters were estimated by sigmoid Emax models. RESULTS: The unbound fraction was significantly diminished in pretreated rats (mean +/- SEM) (6.60 +/- 1.23% vs. control 14.4 +/- 1.40%, P < 0.05). Volume of distribution (V) and clearance for UL and TL were significantly decreased when compared to the controls (P < 0.0001 for UL and P < 0.05 for TL). Plasma clearance based on unbound concentration for UL did not differ between groups but the unbound V and steady-state unbound V remained decreased (P < 0.05 and P < 0.0001). Pretreated rats showed a significantly diminished drug effect: the area under the E-t curve over 180 min was (mean +/- SEM) 5,189 +/- 657.7 in control animals vs. 3,486 +/- 464.4 in the pretreated group (P < 0.05). The EC50 (concentration at half maximum effect) for UL and TL were increased in pretreated rats and were not compensated when the unbound concentration was used. CONCLUSIONS: An increase in AAG causes alterations in the PK and PD of lerisetron, and because this is not compensated with the unbound concentration, we suggest that mechanisms not linked to protein binding may be involved. PMID- 11474790 TI - A novel topoisomerase II poison GL331 preferentially induces DNA cleavage at (C/G)T sites and can cause telomere DNA damage. AB - PURPOSE: Topoisomerase II (Topo II) preferentially cuts DNA at alternating purine pyrimidine repeats. Different Topo II poisons may affect Topo II to produce distinct drug-specific DNA cleavage patterns. GL331 is a new podophyllotoxin derivative exhibiting potent Topo II-poisoning activity. Therefore, the sequence selectivity of GL331-induced DNA cleavage was determined. METHODS: Human gastric adenocarcinoma SC-M1 cells were treated with GL331, and the resultant DNA fragments were isolated by SDS-K+ precipitation. These DNA fragments were further cloned and sequenced to exhibit GL331-induced DNA cleavage sites. In addition, the telomere damage was detected by Southern blot analyses using a (TTAGGG)4 probe. GL331's effect on telomerase was examined using the TRAP assay. RESULTS: The selective sequences of GL331-induced DNA cleavage were analyzed. The first nucleotide 3'-terminal to the cleavage sites was preferentially C or G and followed by the second nucleotide T. More than 50% of GL331-induced DNA cleavage fragments exhibited AT-rich sequences in the first 20 nucleotides. In addition, the telomeric damage was observed both from GL331-treated SC-M1 cells and in vitro incubation of genomic DNA with GL331 and purified human Topo II. Although GL331 treatment reduced cellular telomerase activity, in vitro reaction data suggested that GL331 was not a telomerase inhibitor. CONCLUSION: GL331 preferentially induced Topo II-mediated DNA cleavage at (C/G)T sites. Because the telomeric repeat sequence contains GL331's GT preference site, the telomere was identified as one of the targets of GL331-induced DNA damage. PMID- 11474791 TI - Characterization of two polymorphs of salmeterol xinafoate crystallized from supercritical fluids. AB - PURPOSE: To characterize two polymorphs of salmeterol xinafoate (SX-I and SX-II) produced by supercritical fluid crystallization. METHODS: SX-I and SX-II were crystallized as fine powders using Solution Enhanced Dispersion by Supercritical Fluids (SEDS). The two polymorphs and a reference micronized SX sample (MSX) were characterized using powder X-ray diffractometry (PXRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), aqueous solubility (and dissolution) determination at 5-40 degrees C, BET adsorption analysis, and inverse gas chromatography (IGC). RESULTS: Compared with SX-I, SX-II exhibited a lower enthalpy of fusion, a higher equilibrium solubility, a higher intrinsic dissolution rate, a lower enthalpy of solution (based on van't Hoff solubility plots), and a different FTIR spectrum (reflecting differences in intermolecular hydrogen bonding). Solubility ratio plot yielded a transition temperature (-99 degrees C) below the melting points of both polymorphs. MSX showed essentially the same crystal form as SX-I (confirmed by PXRD and FTIR), but a distinctly different thermal behaviour. Mild trituration of SX-I afforded a similar DSC profile to MSX while prolonged grinding of SX-I gave rise to an endotherm at -109 degrees C, corresponding to solid-solid transition of SX-I to SX-II. Surface analysis of MSX, SX-I, and SX-II by IGC revealed significant differences in surface free energy in terms of both dispersive (nonpolar) interactions and specific (polar) acid-base properties. CONCLUSIONS: The SEDS-processed SX-I and SX-II display high polymorphic purity and distinctly different physical and surface properties. The polymorphs are related enantiotropically with SX-I being the thermodynamically stable form at room temperature. PMID- 11474792 TI - Ritonavir: an extraordinary example of conformational polymorphism. AB - PURPOSE: In the summer of 1998, Norvir semi-solid capsules supplies were threatened as a result of a new much less soluble crystal form of ritonavir. This report provides characterization of the two polymorphs and the structures and hydrogen bonding network for each form. METHODS: Ritonavir polymorphism was investigated using solid state spectroscopy and microscopy techniques including solid state NMR, Near Infrared Spectroscopy, powder X-ray Diffraction and Single crystal X-ray. A sensitive seed detection test was developed. RESULTS: Ritonavir polymorphs were thoroughly characterized and the structures determined. An unusual conformation was found for form II that results in a strong hydrogen bonding network A possible mechanism for heterogeneous nucleation of form II was investigated. CONCLUSIONS: Ritonavir was found to exhibit conformational polymorphism with two unique crystal lattices having significantly different solubility properties. Although the polymorph (form II) corresponding to the "cis" conformation is a more stable packing arrangement, nucleation, even in the presence of form II seeds, is energetically unfavored except in highly supersaturated solutions. The coincidence of a highly supersaturated solution and a probable heterogeneous nucleation by a degradation product resulted in the sudden appearance of the more stable form II polymorph. PMID- 11474793 TI - Fracture property studies of paracetamol single crystals using microindentation techniques. AB - PURPOSE: To study the fracture behavior of the major habit faces of paracetamol single crystals using microindentation techniques and to correlate this with crystal structure and molecular packing. METHODS: Vicker's microindentation techniques were used to measure the hardness and crack lengths. The development of all the major radial cracks was analyzed using the Laugier relationship and fracture toughness values evaluated. RESULTS: Paracetamol single crystals showed severe cracking and fracture around all Vicker's indentations with a limited zone of plastic deformation close to the indent. This is consistent with the material being a highly brittle solid that deforms principally by elastic deformation to fracture rather than by plastic flow. Fracture was associated predominantly with the (010) cleavage plane, but was also observed parallel to other lattice planes including (110), (210) and (100). The cleavage plane (010) had the lowest fracture toughness value, Kc = 0.041MPa m1/2, while the greatest value, Kc = 0.105MPa m1/2 was obtained for the (210) plane. CONCLUSIONS: Paracetamol crystals showed severe cracking and fracture because of the highly brittle nature of the material. The fracture behavior could be explained on the basis of the molecular packing arrangement and the calculated attachment energies across the fracture planes. PMID- 11474794 TI - Compression shear strength and tableting behavior of microcrystalline cellulose agglomerates modulated by a solution binder (polyethylene glycol). AB - PURPOSE: To investigate the possibility of modulating the compression shear strength of agglomerates by the incorporation of a solution binder and to study the subsequent effect on the deformation behavior and tablet forming ability of the agglomerates. METHOD: Various concentrations (0.5 to 10%) of polyethylene glycol were incorporated as a solution binder into microcrystalline cellulose agglomerates of different porosity (10 and 20%) and the shear strength of the agglomerates, as evaluated by the 1/b value of the Kawakita equation, and the permeability to air and tensile strength of tablets formed from them were determined. RESULTS: Increased agglomerate porosity and concentration of polyethylene glycol reduced the 1/b values, which led to the formation of tablets with a lower permeability. A decreased tablet permeability corresponded to an increased tablet tensile strength except that the highest binder content was associated with a drop in the tablet tensile strength. CONCLUSIONS: The solution binder reduced the agglomerate shear strength, which was expressed as an increased degree of agglomerate deformation during compression. The latter seemed to be controlled by both agglomerate porosity and shear strength. The main role of the solution binder in improving the agglomerate compactability was to increase the degree of deformation of agglomerates during compression. PMID- 11474795 TI - Stabilization and controlled release of bovine serum albumin encapsulated in poly(D, L-lactide) and poly(ethylene glycol) microsphere blends. AB - PURPOSE: The acidic microclimate in poly(D, L-lactide-co-glycolide) 50/50 microspheres has been previously demonstrated by our group as the primary instability source of encapsulated bovine serum albumin (BSA). The objectives of this study were to stabilize the encapsulated model protein, BSA, and to achieve continuous protein release by using a blend of: slowly degrading poly(D, L lactide) (PLA), to reduce the production of acidic species during BSA release; and pore-forming poly(ethylene glycol) (PEG), to increase diffusion of BSA and polymer degradation products out of the polymer. METHODS: Microspheres were formulated from blends of PLA (Mw 145,000) and PEG (Mw 10,000 or 35,000) by using an anhydrous oil-in-oil emulsion and solvent extraction (O/O) method. The polymer blend composition and phase miscibility were examined by FTIR and DSC, respectively. Microsphere surface morphology, water uptake, and BSA release kinetics were also investigated. The stability of BSA encapsulated in microspheres was examined by losses in protein solubility, SDS-PAGE, IEF, CD, and fluorescence spectroscopy, RESULTS: PEG was successfully incorporated in PLA microspheres and shown to possess partial miscibility with PLA. A protein loading level of 5% (w/w) was attained in PLA/PEG microspheres with a mean diameter of approximately 100 microm. When PEG content was less than 20% in the blend, incomplete release of BSA was observed with the formation of insoluble, and primarily non-covalent aggregates. When 20%-30% PEG was incorporated in the blend formulation, in vitro continuous protein release over 29 days was exhibited. Unreleased BSA in these formulations was water-soluble and structurally intact. CONCLUSIONS: Stabilization and controlled relaease of BSA from PLA/PEG microspheres was achieved due to low acid and high water content in the blend formulation. PMID- 11474796 TI - Enantioseparation of flobufen with cyclodextrins studied by capillary electrophoresis and NMR. AB - PURPOSE: The aim of this study was to obtain the resolution of flobufen enantiomers, an antiinflammatory active substance, by capillary electrophoresis with cyclodextrins. The mechanism of complexation and determination of the stoichiometry of the complexes were studied by NMR and the analytical method was developed and validated. METHODS: Zone capillary electrophoresis coupled to direct ultraviolet detection was selected. The interaction between flobufen and the chiral selector was studied by NMR. Optimization of the separation was performed using a Box-Wilson Central Composite Design for three factors related to the composition of the electrolyte. RESULTS: Heptakis (2,3,6-tri-O-methyl) beta-cyclodextrin (TM-beta-CD) was found to be the most efficient selector via the formation of a 1:1 complex proved by NMR. Constants of complexation of flobufen enantiomers were determined by NMR and capillary electrophoresis. Optimal values for the critical factors of the analytical system were: pH (5.50), content in methanol (10% v/v), and TM-beta-CD (30 mM). The ability of capillary electrophoresis to quantify as low as 0.1% (w/w) of R in S-flobufen or vice-versa was established. CONCLUSIONS: Capillary electrophoresis was shown to be a valuable method to control the enantiocomposition of flobufen by use of a chiral selector whose interactions with the analytes could be explored by NMR. PMID- 11474797 TI - Magainin-mediated disruption of stratum corneum lipid vesicles. PMID- 11474798 TI - Neural network classification of nerve activity recorded in a mixed nerve. AB - Whole-nerve cuff electrodes can be used to record electrical nerve activity in peripheral nerves and are suitable for chronic implantation in animals or humans. If the whole nerve innervates multiple target organs or muscles then the recorded activity will be the superposition of the activity of different nerve fibers innervating these organs. In certain cases it is desirable to monitor mixed nerve activity and to determine the origin (modality) of the recorded activity. A method using the autocorrelation function of recorded nerve activity and an artificial neural network was developed to classify the modality of nerve signals. The method works in cases where different end organs are innervated by nerve fibers having different diameter distributions. The electrical activity in the cat S1 sacral spinal root was recorded using a cuff electrode during the activation of cutaneous, bladder, and rectal mechanoreceptors. Using the classification method, 87.5% of nerve signals were correctly classified. This result demonstrates the effectiveness of the neural network classification method to determine the modality of the nerve activity arising from activation of different receptors. PMID- 11474799 TI - Estimation of intra-cranial neural activities by means of regularized neural network-based inversion techniques. AB - Artificial neural networks can be exploited to solve inverse problems arising from the estimation of neural activities in the brain. In this paper, we review the network inversion techniques for solving inverse problems with special attention directed towards electroencephalographic dipole localization and the improvement of positron emission tomography. In our regularized network inversion technique, for stabilizing the solution, we explicitly include the a priori knowledge by adding penalty terms to the energy function and/or build this knowledge into the architecture of the multi-layered neural networks that are used as an inverse problem solver. In the electroencephalogram analysis, the consensus term added to the energy function facilitated 3-dipole localization for visually evoked potentials. Effectiveness of our regularization is shown in improving the positron emission tomographic images and for generating metabolic images of the brain, under the constraints given by the a priori knowledge inherent to the measurement systems and physiological rules. PMID- 11474800 TI - Cortical spreading depression and the pathogenesis of brain disorders: a computational and neural network-based investigation. AB - This paper reviews our recent studies of the role of cortical spreading depression (CSD) in the pathogenesis of brain disorders. Our investigation is a computational one, involving the development and utilization of a complex neuro metabolic model of the interactions assumed to occur in the cortex during the passage of multiple CSD waves. Incorporating these neuro-metabolic changes of CSD within a neural network model of normoxic cortex produces cortical activation patterns during the passage of a CSD wave that, projected onto the visual fields, resemble the visual hallucinations observed during the migraine aura. When focal ischemia is simulated with the model, the evoked CSD waves are found to affect the expansion of the infarction into the ischemic penumbra. Our findings support the hypothesis that CSD does play an important pathogenic role in these and other neurological disorders, and suggest additional experimental studies that may further substantiate it. PMID- 11474801 TI - EEG source localization: a neural network approach. AB - Functional activity in the brain is associated with the generation of currents and resultant voltages which may be observed on the scalp as the electroencephelogram. The current sources may be modeled as dipoles. The properties of the current dipole sources may be studied by solving either the forward or inverse problems. The forward problem utilizes a volume conductor model for the head, in which the potentials on the conductor surface are computed based on an assumed current dipole at an arbitrary location, orientation, and strength. In the inverse problem, on the other hand, a current dipole, or a group of dipoles, is identified based on the observed EEG. Both the forward and inverse problems are typically solved by numerical procedures, such as a boundary element method and an optimization algorithm. These approaches are highly time-consuming and unsuitable for the rapid evaluation of brain function. In this paper we present a different approach to these problems based on machine learning. We solve both problems using artificial neural networks which are trained off-line using back-propagation techniques to learn the complex source-potential relationships of head volume conduction. Once trained, these networks are able to generalize their knowledge to localize functional activity within the brain in a computationally efficient manner. PMID- 11474802 TI - The callosal dilemma: explaining diaschisis in the context of hemispheric rivalry via a neural network model. AB - It is often suggested that a major factor in diaschisis is the loss of transcallosal excitation to the intact hemisphere from the lesioned one. However, there is long-standing disagreement in the broader experimental literature about whether transcallosal interhemispheric influences in the human brain are primarily excitatory or inhibitory. Some experimental data are apparently better explained by assuming inhibitory callosal influences. Past neural network models attempting to explore this issue have encountered the same dilemma: in intact models, inhibitory callosal influences best explain strong cerebral lateralization like that occurring with language, but in lesioned models, excitatory callosal influences best explain experimentally observed hemispheric activation patterns following brain damage. We have now developed a single neural network model that can account for both types of data, i.e., both diaschisis and strong hemisphere specialization in the normal brain, by combining excitatory callosal influences with subcortical cross-midline inhibitory interactions. The results suggest that subcortical competitive processes may be a more important factor in cerebral specialization than is generally recognized. PMID- 11474803 TI - Control of neuromuscular stimulation for ambulation by complete paraplegics via artificial neural networks. AB - The paper describes the application of a neural network (ANN) for controlling a functional neuromuscular stimulation (FNS) system to facilitate patient responsive ambulation by paralyzed patients with traumatic, thoracic-level spinal cord injuries. The particular ANN that is employed is a modified Adaptive Resonance-Theory (ART-1) network. It serves as a controller in an FNS system (the Parastep system) that is presently in use by approximately 500 patients worldwide (but still without ANN control) and which was the first and only FNS system approved by FDA. The proposed neural network discriminates above-lesion upper trunk electromyographic (EMG) time series to activate standing and walking functions under FNS and controls FNS stimuli levels using response-EMG signals. For this particular application, several modifications are introduced into the standard ART-1 ANN. First, a modified on-line learning rule is proposed. The new rule assures bi-directional modification of the stored patterns and prevents noise interference. Second, a new reset rule is proposed, which prevents 'exact matching' when the input is a subset of the chosen pattern. A single ART-1-based structure is being applied to solving two problems, namely (1) signal pattern recognition and limb function determination, and (2) control of stimulation levels. This also facilitates ambulation of paraplegics under FNS, with adequate patient interaction in initial system training, retraining the network when needed, and in allowing patient's manual over-ride in the case of error, where any manual over-ride serves as a re-training input to the neural network. The ANN control facilitates continuous update of control settings during normal use, without formal retraining. PMID- 11474804 TI - On the inter-relations between artificial and physiological neural networks. AB - This paper discusses the inter-relations between findings on the physiological neural network (PNN) and artificial neural networks (ANN). It discusses the interaction of progress in both PNN and ANN for the purpose of borrowing from ANN's mathematical understandings to establish pointers for further explorations to better understand the PNN, and also for the reciprocal transferring of knowledge from PNN findings to improve ANN schemes. Such improvements in ANN are essential for better handling the needs of the information technology (IT) explosion in dealing with huge data bases and where data often defy analysis and are incomplete and fuzzy. On the other hand, principles and elements of ANN designs that appear to be important and successful can serve as guides for identifying them in the PNN, to be subsequently confirmed by bioanalytical tests. Hence progress in PNN is obviously essential for progress in ANN, as is progress in ANN helpful in PNN modeling, though its laboratory confirmation is still a far lengthier process. We discuss certain specific ANN schemes with respect to the above inter-relations with PNN. We feel that the progress in both PNN and ANN research provides a major link between the thrust in information technology developments and the thrust in biological science research, which are most probably the two major focus areas of research at the dawn of the 21st century. PMID- 11474805 TI - Neural models for auditory localization based on spectral cues. AB - In this paper we analyze several auditory localization neural models that are based on head related transfer functions (HRTFs). HRTFs represent the combined directional-spectral response of the pinnae head and torso. The role of HRTFs in such modeling has hitherto been underestimated despite substantial experimental evidence to its relevance in spatial hearing, especially in determining direction of high-frequency sound sources. In the first section we suggest a neural model that links the physiology of binaural processing to a neural network that extracts spectral ratios. These ratios correspond to HRTFs ratios and can provide auditory directional cues. Next, we compare several methods of matching HRTFs ratios using discriminative matching measure (DMM). We consider several solutions to the matching problem from a neural signal processing viewpoint. We compare correlation based approaches with DMM optimization approach and with a non-linear approach based on neural back-propagation algorithm. All three models can be implemented by neural networks. Finally, we include experimental results of simulations that are conducted using these methods. Experiments show that the back-propagation based neural network yields the best results in terms of DMM both for narrow-band and broad band excitation. The back-propagation neural network is also superior in matching noisy HRTF ratio vectors. PMID- 11474806 TI - Effects of QiGong on brain function. AB - QiGong is an ancient and widely practiced Chinese meditation exercise. We studied the effects of QiGong on brain function with modern neuromonitoring tools in two subjects. In a male QiGong master (extremely trained practitioner), the technique induced reproducible changes in transcranial Doppler sonography, EEG, stimulus induced 40 Hz oscillations, and near-infrared spectroscopy findings. Similar effects were seen after the application of multimodal stimuli and when the master concentrated on intense imagined stimuli (e.g. 22.2% increase in mean blood flow velocity (vm) in the posterior cerebral artery, and a simultaneous 23.1% decrease of vm in the middle cerebral artery). Similar effects were seen in the female subject. Neuromonitoring during QiGong appears able to objectify accompanied cerebral modulations surrounding this old Chinese meditation exercise. PMID- 11474807 TI - Physiological levels of beta-amyloid induce cerebral vessel dysfunction and reduce endothelial nitric oxide production. AB - beta-amyloid (A beta), the major component of senile plaques in Alzheimer's disease (AD), normally circulates in the blood at nanomolar levels but is elevated in AD. Previous studies have found that high concentrations (10(-5)-10( 4) M) of A beta result in neuronal cell death. Here we show that physiological levels of soluble A beta can induce dysfunction in perfused rat cerebral vessels and in cultured endothelial cells. At concentrations of 10(-9)-10(-6) M, A beta induced a significant concentration-dependent reduction of NO production in endothelial cells. At 10(-8) M, A beta significantly decreased the sensitivity of cerebral vessels to acetylcholine (ACh), an endothelium dependent vasodilator. At 10(-7) M and higher concentrations, A beta significantly reduced the maximum response of vessels to ACh, and induced significant endothelial cell death. A beta (10(-9)-10(-5) M) did not cause any detectable change in nitric oxide synthase levels. The results suggest that a modest increase in the concentration of A beta above its normal physiological level in the circulation, as found in the early stages of AD, results in decreased NO production and vessel sensitivity to endothelium-dependent vasodilation that could lead to constricted blood vessels and ischemia in the surrounding tissue. Further increases in A beta concentration, which may occur in the later stages of AD, result in cell death and decreased maximum vasodilator response of cerebral vessels. PMID- 11474808 TI - Pericapillary arteriovenous malformations angiographically manifested as cerebral venous malformations. AB - Cerebral venous malformations have been diagnosed by angiographic features and are considered to be a benign anomaly. However, ample evidence indicates that stroke or similar symptomatology occurs in patients harboring a cerebral vascular malformation that was diagnosed angiographically as a venous malformation. The purpose of the study is to confirm the presence of a pericapillary arteriovenous malformation in these patients by analyzing the clinical history and surgical findings and correlating them with histological features. Thirteen patients were included in this study. Each patient fulfilled four criteria: 1. the patient was neurologically symptomatic; 2. the angiographic diagnosis was a venous malformation; 3. at operation, shunting arterioles (50-100 microns) were found to contribute to the malformation; and 4. histologically, a mixture of venous channels and arterioles with arterioles directly connected to venules was found. Based on the above findings, the malformation present in the 13 patients can be termed a 'pericapillary arteriovenous malformation'. Its angiographic distinction from the cerebral venous malformation requires technological advancement in the capability of magnifying images of arterioles and venules, along with improvement in image resolution. PMID- 11474809 TI - Crucial role of calpain in hypoxic PC12 cell death: calpain, but not caspases, mediates degradation of cytoskeletal proteins and protein kinase C-alpha and delta. AB - Ca2+ influx is one of the main causative events in hypoxic PC12 cell death, because an extracellular Ca2+ chelator, ethylene glycol bis (2-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) inhibited and Ca2+ ionophore A23187 mimicked the hypoxic cell death. The hypoxic cell death was markedly prevented by a broad spectrum caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z VAD-FMK) as well as a calpain inhibitor, calpeptin, as assessed by nuclear staining with Hoechst 33258 and lactate dehydrogenase release. The processing of procaspase-3 was inhibited by z-VAD-FMK, but not by calpeptin. In contrast, z-VAD FMK failed to block the proteolytic cleavage of fodrin-alpha, a preferential substrate for calpain. On the other hand, degradation of actin and fodrin-alpha was prevented by calpeptin but not by z-VAD-FMK. In addition, not only protein kinase C (PKC)-alpha but also PKC-delta were cleaved to generate approximately 46 kDa fragments. The PKC fragmentation was inhibited by calpeptin but not by z-VAD FMK. These findings suggest that the extracellular Ca2+ influx induced by hypoxic stress activates calpain, resulting in the degradation of cytoskeletal proteins and generation of PKC fragments almost independently of caspase activation. Therefore, calpain may play an important role in hypoxic PC12 cell death. PMID- 11474810 TI - Extracellular levels of glucose and lactate measured by quantitative microdialysis in the human brain. AB - The aim of this study was to use quantitative microdialysis to estimate the true extracellular concentrations of glucose and lactate in minimally disturbed human brain. These values are important as criteria for microdialytical monitoring in critical care patients and for determining therapy. Microdialysis procedures were carried out during tumor operations, the probe being inserted distant from the site of manipulation in minimally disturbed tissue. Two methods were used: 1. The zero net flux method of Lonnroth. 2. The low flow method (10 mm membrane length, flow rate 0.3 microliter min-1, high in vivo recovery). Both methods gave similar values of about 2000 microM for lactate and slightly less for glucose (1700 microM). Glucose levels correspond with those measured by other methods in humans, allowing for the fact that our patients were anesthetised. Extracellular glucose levels were positively correlated with blood glucose values measured before the operation, and with extracellular lactate. Results confirm that extracellular glucose is zero when blood glucose is about 2 mM. PMID- 11474811 TI - Pre-clinical subdural tissue reaction and absorption study of absorbable hemostatic devices. AB - SURGIFOAM (Absorbable Gelatin Sponge, USP), a new absorbable hemostatic sponge, GELFOAM (Absorbable Gelatin Sponge, USP) or Avitene (microfibrillar collagen hemostat) were evaluated in a three-month tissue reaction and absorption study in rabbits. Bilateral craniotomy was followed by subdural implantation of each hemostatic device. A sham control group was treated in a similar way except that no material was implanted. Implantation of these hemostatic devices for 15, 43, or 92 days did not result in any deaths or clinical neurobehavioral abnormalities, changes in cerebrospinal fluid, or significant macroscopic observations at necropsy. The tissue reaction to SURGIFOAM sponge was characterized by transient granulomatous inflammation that was slightly less intense than that observed for GELFOAM sponge which correlated to slightly longer absorption. In contrast, the tissue reaction to Avitene hemostat was characterized by moderate to marked granulomatous inflammation with an acute inflammatory component indicating a greater degree of tissue irritancy. Sequelae of this reaction were still observed at 92 days post-implantation. The tissue reaction in humans to SURGIFOAM sponge used as a hemostatic agent for neurologic surgical procedures is expected to be comparable to that observed with GELFOAM sponge, resulting in no significant adverse reactions for patients. This animal model was useful to assess the tissue reaction and absorption of biomaterials implanted in contact with the central nervous system, and it was able to differentiate between materials of biologic origin. PMID- 11474812 TI - Gene therapy for treatment of cerebral ischemia using defective herpes simplex viral vectors. AB - Significant advances have been made over the past few years concerning the cellular and molecular events underlying neuron death. Recently, it is becoming increasingly clear that some of the genes induced during cerebral ischemia may actually serve to rescue the cell from death. However, the injured cell may not be capable of expressing protein at levels high enough to be protective. One of the most exciting arenas of such interventions is the use of viral vectors to deliver potentially neuroprotective genes at high levels. Neurotrophic herpes simplex viral strains are an obvious choice for gene therapy to the brain, and we have utilized bipromoter vectors that are capable of transferring various genes to neurons. Using this system in experimental models of stroke, cardiac arrest and excitotoxicity, we have found that it is possible to enhance neuron survival against such cerebral insults by over-expressing genes that target various facets of injury. These include energy restoration by the glucose transporter (GLUT-1), buffering calcium excess by calbindin, preventing protein malfolding or aggregation by stress proteins and inhibiting apoptotic death by BCL-2. We show that in some cases, gene therapy is also effective after the onset of injury, and also address whether successful gene therapy necessarily spares function. Although gene therapy is limited to the few hundred cells the vector is capable of transfecting, we consider the possibility of such gene therapy becoming relevant to clinical neurology in the future. PMID- 11474813 TI - Inflated perception of responsibility for harm in obsessive-compulsive disorder. AB - The present study examined the proposition that inflated responsibility is implicated in obsessive compulsive disorder (OCD). Compared to non-anxious control participants (NACs), and an anxious control group with generalized social phobia (GSPs), we predicted that individuals with OCD (OCs) would exhibit a greater urge to rectify situations involving potential risk, would report more distress upon leaving such situations unrectified, and would feel more personal responsibility if the unrectified situations resulted in harm. Fifteen OCs, 15 NACs, and 15 GSPs completed the Obsessive Compulsive Responsibility Scale (OCRS), which included low-risk, OC-relevant, and high-risk situations. Compared to NACs and GSPs, OCs reported more urges, distress, and responsibility in low-risk and OC-relevant situations; no group differences were detected on high-risk situations. GSPs and NACs differed only in their responsibility in OC-relevant situations, with GSPs reporting higher responsibility than controls. Our results suggest that compared to OCs, NACs, and GSPs can better differentiate between situations that merit concern and ones that do not. PMID- 11474814 TI - Cognitive-behavior therapy of obsessive-compulsive disorder in private practice: an effectiveness study. AB - A controversy exists over whether or not the results of randomized controlled trials (RCTs) are generalizable to routine clinical practice. The present study examines the effectiveness of cognitive-behavior therapy (CBT) for obsessive compulsive disorder (OCD) in a private practice setting. Twenty-six consecutive clients referred to a private anxiety disorders specialty clinic began treatment for OCD. Of the 19 (73%) clients who completed treatment, 84% were treatment responders. Clients, treatment, and outcome of the present study are compared with those of representative RCTs, and it is concluded that there are more similarities than differences. It is concluded that CBT can be effectively delivered in routine clinical practice. PMID- 11474815 TI - The Claustrophobia Questionnaire. AB - The content and psychometric properties of the Claustrophobia Questionnaire (CLQ) are described. An earlier version of the CLQ was developed to test the hypothesis that claustrophobia is comprised of two distinct but related fears--the fear of suffocation and the fear of restriction [J. Anxiety Disord. 7 (1993) 281.]. The scale was used to assess patients undergoing the magnetic resonance imaging (MRI) procedure [J. Behav. Med. 21 (1998) 255.] and in participants with panic disorder [J. Abnorm. Psychol. 105 (1996) 146; Taylor, S., Rachman, S., & Radomsky, A. S. (1996). The prediction of panic: a comparison of suffocation false alarm and cognitive theories. Unpublished data.]. On the basis of these studies, we decided to revise and shorten the CLQ, collect normative data, and provide information on the scale's predictive and discriminant validity as well as its internal consistency and test-retest reliability. This was done through a set of four interconnected studies that included psychometric analyses of undergraduate and community adult questionnaire responses and behavioural testing. Results indicate that the CLQ has good predictive and discriminant validity as well as good internal consistency and test-retest reliability. The CLQ appears to be a reliable and sensitive measure of claustrophobia and its component fears. We encourage the use of the CLQ in a variety of clinical and research applications. The scale is provided in this paper for public use. PMID- 11474816 TI - Selective attentional biases for physical threat in physical anxiety sensitivity. AB - Anxiety sensitivity is a trait susceptibility associated with the fear of anxiety related sensations. One reason why such fears exist may be because those high in anxiety sensitivity selectively attend towards such sensations. However, few studies have actually investigated these cognitive biases in high anxiety sensitive individuals. The current study, therefore, sought to investigate selective attentional biases using the visual dot-probe paradigm. Since recent research suggests that at least one component of anxiety sensitivity is linked to the fear of physical sensations, individual were selected on the basis as to whether they were high or low in their anxious concern for physical sensations. In order to determine whether a general or specific attentional bias exists, the emotionality of material presented to participants was varied in terms of whether it was physically threat-related, socially threat-related, or positive. Consistent with predictions, those high in physical anxiety sensitivity were found to exhibit a selective attentional bias in favour of the location of physically threatening material. Furthermore, those low in anxiety sensitivity were found to avoid such material. Interestingly, a similar attentional bias was not found for either socially threatening or positive material. If anything, those high in physical anxiety sensitivity avoided positive material. These findings are discussed in light of current theories of anxiety sensitivity and future research. PMID- 11474817 TI - Psychopathological correlates of anxiety sensitivity: evidence from clinical interviews and self-report measures. AB - Most research on anxiety sensitivity (AS) and its relation to psychopathology has examined the Anxiety Sensitivity Index (ASI) in various clinical samples. The present study was the first to investigate psychopathological correlates of AS using self-report measures, the anxiety, somatoform, and substance use disorders sections of the Structured Clinical Interview for the DSM-IV (SCID), and open ended interview questions about the subjective meaning and origins of AS. A college student sample (N=317) was used for the correlational analysis, and subsamples of high-AS (n=44) and low-AS (n=41) participants completed the interviews. Despite having an ASI score (Mean=35.0) that was equivalent to that observed in panic disorder samples, the screening questions in the clinical interview indicated that only 55% of high-AS participants had panic attacks and the diagnostic assessment indicated that only 30% met criteria for an anxiety disorder. Several findings suggested that AS, at least as measured by the ASI, was strongly related to anxiety disorder symptoms and diagnoses, although there was substantial variation within the anxiety disorder classifications. There was also evidence that AS might extend to a broader catastrophic style concerning bodily symptoms and health that go beyond anxiety symptoms per se. The subjective meaning of AS derived from the interview data stressed the need for a formal, structured interview of AS. PMID- 11474818 TI - Disability measurement in the anxiety disorders: comparison of three brief measures. AB - General measures of disablement are powerful tools in clinical settings as they provide a link between clinic and community populations and allow cross-disorder comparisons. Because of their generic nature, they allow the transmittal of comprehensible health planning information to decision makers. We located no studies of such general disability measures in the anxiety disorders and decided to examine the properties of three brief generic measures in an anxiety disorders clinic. Consecutive attenders (N=168) were administered the Medical Outcomes Study Short Form-12 item (SF-12), the Brief Disability Questionnaire (BDQ), and a measure of the number of disability days due to health over the past 4 weeks. These measures were compared on their ability to discriminate within the clinic and in comparison to a representative community group. The mental health scale of the SF-12 was the most sensitive of the generic measures to differences in functioning, and is recommended as a measure of such for people with anxiety disorders. Attenders at this anxiety clinic reported high levels of disablement, and normative data on the general measures are provided. PMID- 11474819 TI - The structure of posttraumatic stress disorder symptoms in combat veterans: a confirmatory factor analysis of the impact of event scale. AB - There has been controversy over the most appropriate way to define symptom clusters for posttraumatic stress disorder (PTSD). We tested the factor structure of the Impact of Event Scale (IES) in a sample of 195 male combat veterans with chronic PTSD by using confirmatory factor analysis. The two-factor model including Intrusion (i.e., unwanted memories of the event) and Avoidance (i.e., attempts to avoid reminders and numbing of emotional responsiveness) deviated significantly from good fit. However, a four-factor model, including Intrusion and Effortful Avoidance subscales, as well as Sleep Disturbance and Emotional Numbing subscales, fit significantly better. Correlations with other PTSD measures are explored and implications for the conceptualization of PTSD are discussed. PMID- 11474820 TI - Efficacy of Bach-flower remedies in test anxiety: a double-blind, placebo controlled, randomized trial with partial crossover. AB - Bach-flower remedies are a type of alternative medication used increasingly for over-the-counter self-help purposes. We studied the efficacy of a combination of Bach-flower remedies in subjects with test anxiety in a randomized, placebo controlled, blinded parallel group design, with crossing over the placebo group to remedies after the first phase. Anxiety was measured by a standardized, validated test anxiety questionnaire (the German version of the Test Anxiety Inventory, TAI-G). Fifty-five of 61 subjects with self-reported test anxiety gave valid data. There was no significant difference between the groups, but a significant decrease of test anxiety in all groups was present. We conclude that Bach-flower remedies are an effective placebo for test anxiety and do not have a specific effect. PMID- 11474823 TI - The promise of optical sensing strategies for glucose. PMID- 11474824 TI - Current activities at the Centers for Disease Control and Prevention's National Diabetes Laboratory. AB - In 1997, the Centers for Disease Control and Prevention established the National Diabetes Laboratory in order to help prevent and treat type 1 diabetes. This state-of-the-art laboratory collaborates with research scientists and key national and international organizations throughout the world to identify and study risk factors for type 1 diabetes by developing measurements for glycosylated proteins, developing and evaluating technology for measuring genetic risk factors for the disease, and working to standardize autoantibody measurements. Developing improved technologies for diagnosing and managing diabetes and developing reference materials for properly calibrating and standardizing blood glucose meters are also critical aspects of the laboratory's work. In addition, the laboratory provides quality storage for valuable collections of biologics and other materials and facilitates sharing of specimens, associated epidemiologic data, and test results. Working with our partners in diabetes research, we are improving the diagnosis, treatment, and prevention of type 1 diabetes. PMID- 11474825 TI - Prospective evaluation of accuracy, precision, and reproducibility of an at-home hemoglobin A1c sampling kit. AB - BACKGROUND: The measurement of hemoglobin A1c (HbA1c) is critical to the optimal therapeutic management of diabetes. To be most useful, the HbA1c value should be available at the clinical visit. Recently, a number of at-home sampling kits have been developed that facilitate the timely availability of HbA1c results. This is a report of the accuracy, precision (among-subject), and reproducibility (within subject) of one such kit, B-D A1c, At-Home-Test, which combines a filter paper technique for spotting capillary blood with an immunoturbidometric assay (a variation of the Cobas Integra Hemoglobin A1c method also referred to as Roche Unimate). METHODS: The B-D A1c At-Home test kit was evaluated in a clinical trial using 1625 dried blood spot samples from 59 subjects diagnosed with type 1 or type 2 diabetes collected in an in-clinic setting. Data for replicate samples were compared against those from the standard Cobas Integra Hemoglobin A1c assay and from the BioRad Variant high-performance liquid chromatography [HPLC] assay. The effects of subjecting the dried spotted blood samples to prolonged elevated temperatures was evaluated in a separate laboratory analysis. RESULTS: The B-D A1c At-Home results, which are highly correlated with the standard Cobas Integra Hemoglobin A1c assay, (r2 = 94.7%), demonstrate excellent within-subject reproducibility for 3- to 10-day-old samples (coefficient of variation = 2.7%), and provide a coefficient of variation for among-subjects that is 3.9%. There were no clinically significant differences (i.e., < 0.3 units) in samples aged 3 to 10 days, between venous or capillary blood samples, or from freezing and thawing or prolonged exposure of B-D A1c At-Home dried blood samples to elevated temperatures before assay. CONCLUSIONS: The B-D A1c At-Home kit combines the accuracy, precision, and reproducibility of a clinical laboratory test with the convenience of at-home sample collection. This product may add to the convenience of both patient and health care provider by making it easier for patients to obtain their HbA1c values and have them available at their visit to the clinician. PMID- 11474826 TI - Glycohemoglobin testing in patients with diabetes: it may be underutilized now, but that is certain to change. PMID- 11474827 TI - Glycosal: the first rapid, point-of-care test for the determination of hemoglobin A1c in patients with diabetes. AB - BACKGROUND: Glycosal is a rapid, point-of-care test using the established method of affinity chromatography for the quantitative determination of hemoglobin A1c (HbA1c). Glycosal enables the measurement of HbA1c to be performed in the diabetic clinic or doctor's office during a normal consultation, within minutes. The Glycosal test cartridge integrates with HaemaQuant, a compact spectrophotometric analyser equipped with a light emitting diode (LED) photo optic detection system. An interactive liquid crystal display (LCD) provides the operator with an intuitive guide through the test procedure with the final result being expressed as HbA1c. Glycosal has a linear working range of at least 3%-18% HbA1c, requires approximately 10 microL of whole blood and generates a result in approximately 4 minutes. Glycosal has a mean coefficient of variation of less than 5% across its linear range. METHODS: Glycosal was evaluated in the clinical environment by means of comparison to two routine HbA1c autoanalyser methods used in the Clinical Chemistry Department of the Malmo University Hospital, Sweden. The Variant (BioRad) and the Mono-S (Pharmacia) systems are both cation exchange high-performance liquid chromatography (HPLC) systems specifically measuring HbA1c. Fresh (< 2 days) EDTA anticoagulated blood samples were randomly selected from the routine diabetic clinic (n = 54) along with nondiabetic samples from consenting volunteers (n = 8). RESULTS: The Variant and Mono-S HPLC methods showed excellent agreement compared to each other (correlation coefficient [r] > 0.99). The agreement between Glycosal and Variant (r = 0.98) and Glycosal and Mono-S (r > 0.98) HPLC methods were almost equally as significant. A further final evaluation of a 10 sample series (4%-10% HbA1c) of the hospital's HbA1c reference materials by Glycosal produced very strong agreement (r > 0.99) against the Mono-S HPLC. CONCLUSIONS: The Glycosal rapid test is proven to be comparable to two high-precision cation exchange HPLC autoanalyzers and constitutes a low cost, HbA1c measurement system for the management of individuals suffering from diabetes mellitus at the point-of-care. PMID- 11474828 TI - Glycosal. Commentary. PMID- 11474829 TI - Clinical validity of a self-test fructosamine in outpatient diabetic management. AB - BACKGROUND: Measurement of glycated hemoglobin and hemoglobin A1c (HbA1c) is now well established as the best means of measuring overall glucose control in managing diabetes. Other glycated serum protein assays reflecting recent glycemic control, e.g., glycated albumin and glycated protein (fructosamine), have also been validated in clinical studies. Regardless of the method, the expense and inconvenience of laboratory testing of blood samples may contribute to the well documented underutilization of clinical glycated protein assessment. Accordingly, a rapid, inexpensive fingerstick test of fructosamine has been developed. This study cross-sectionally and prospectively assesses the clinical validity of fingerstick fructosamine versus laboratory determination. METHODS: Fifty-one subjects (18 control, 33 with diabetes) participated in a cross-sectional study and 20 subjects with type 2 diabetes participated in a prospective, 6-week study with clinical intervention consisting of glipizide gits or metformin in mono- and combination therapy. Subjects had weekly laboratory determination of serum fructosamine, glycated hemoglobin, and fasting glucose; fingerstick fructosamine was obtained at each clinic visit. RESULTS: Fingerstick fructosamine was shown to correlate highly to laboratory fructosamine (r = 0.80, p < 0.001) and glycated hemoglobin (r = 0.81, p < 0.001). In the clinical intervention study subjects, significant decreases in fasting glucose (p < 0.001), laboratory fructosamine (p < 0.001), and fingerstick fructosamine (p < 0.001) were noted compared to baseline. The subject's self-test fingerstick fructosamine mirrored laboratory testing of fructosamine in detection of changes in clinical glucose control. CONCLUSIONS: This study demonstrates that fingerstick fructosamine correlates well to laboratory assessment of fructosamine and glycated hemoglobin. The patient self-test fructosamine provides the same clinical information as laboratory assessment. PMID- 11474830 TI - Fructosamine, hemoglobin A1c, and measures of diabetic control. PMID- 11474831 TI - Determination of reference values for a novel ketoamine-specific fructosamine assay for assessment of diabetic glycemic control. AB - BACKGROUND: Intensive monitoring to improve glycemic control is essential for effective management of diabetes and reduction of long-term morbidity and pathology. Measurement of glycated serum proteins (fructosamine) allows more frequent assessment (monthly) of glycemic control than the 2- to 3-month window of the traditional glycated hemoglobin (HbA1c) assay. In response to concerns about assays designed to measure glycated serum proteins based on the nitroblue tetrazolium (NBT) methodology, a novel assay to measure glycated serum proteins has been developed based on the specific oxidation of the ketoamine bonds within the glycated proteins. METHODS: Reference range values for this new, enzymatic glycated-serum-protein assay (GlyPro; Genzyme Corporation, Cambridge, MA) are reported. RESULTS: The GlyPro reference range is lower and shows close correlation with ranges reported for the NBT assay. The 95% overall reference range was 122 to 236 mumol/L. CONCLUSIONS: GlyPro is a reliable, accurate assay and correlates well with the NBT assay for the measurement of glycated serum proteins. The assay may be useful in the short-term assessment of diabetes control, a necessary complement to long-term control as assessed by hemoglobin A1c (HbA1c) assays. PMID- 11474832 TI - Fructosamine, III: Prologue or epitaph? PMID- 11474833 TI - Fructosamine monitoring in patients with diabetes: is there a need? PMID- 11474834 TI - Comparison of a radioimmunoassay with a microparticle enzyme immunoassay of insulin for use with the minimal model method of determining whole-body insulin sensitivity. AB - The purpose of this study was to compare minimal model results of insulin sensitivity and glucose effectiveness using insulin levels measured by a conventional radioimmunoassay (RIA) versus an automated microparticle enzyme immunoassay (MEIA). Thirty obese subjects participated in an insulin-modified frequently sampled intravenous glucose tolerance test. The MEIA exhibited lower day-to-day variability than did the RIA. The MEIA yielded lower insulin values compared to the RIA probably because of the high cross-reactivity with proinsulin in the RIA. The MEIA yielded a good correlation with the RIA for both insulin sensitivity (r = 0.97, p = 0.0001) and for glucose effectiveness (r = 0.98, p = 0.0001). The MEIA did not significantly effect the results of the MinMod analysis and the low cross-reactivity with proinsulin makes MEIA preferable when insulin sensitivity (SI) is measured in patients with diabetes or obese individuals whose insulin:proinsulin ratio is altered. PMID- 11474835 TI - Microelectromechanical systems technology to deliver insulin. AB - A new microfabrication technology, microelectromechanical systems (MEMS), is envisioned for improved insulin delivery in the context of a device currently being developed for the Defense Advanced Research Projects Agency (DARPA). The drug delivery system utilizes MEMS technology to move and control fluids at the microscale, making possible the reconstitution and delivery of extremely small amounts of drug with extreme precision. In this article, the required microscale components that are currently being developed for the system are described. MEMS are made using fabrication methods similar to that utilized in microelectronics. Consequently, MEMS technology can be used to fabricate devices that are extremely small. The fundamental difference is that MEMS devices can either move themselves or control the movement of other materials, such as fluids. Furthermore, this manufacturing method is intrinsically low-cost and therefore is ideal for drug delivery systems. The current development of a new drug delivery system for controlled drug reconstitution and delivery system for DARPA is described as are the MEMS-based components for the required fluidic control. The adaptation of the system for insulin delivery is addressed and is envisioned to be a fully self contained parenteral drug delivery system about the size of a 4-mm thick credit card. PMID- 11474836 TI - Diabetic nephropathy. AB - Renal involvement is one of the major microvascular complications of both type 1 and type 2 diabetes mellitus. Diabetic nephropathy is the major cause of end stage renal failure in most Western nations and is associated with increased morbidity and mortality as compared to other causes of renal disease. The pathogenesis of renal involvement in diabetes is presumed to be the result of the interplay of metabolic and hemodynamic factors. Significant advances in the prevention and treatment of progression of diabetic nephropathy have been achieved with intensive glycemic control and the treatment of elevated blood pressure. Patients with diabetes should thus be screened regularly for the appearance of any of the risk factors for renal or other complications and treated intensively according to established guidelines for control of hyperglycemia and hypertension. Ancillary therapeutic measures include treatment of hyperlipidemia, low-protein diet, and the cessation of smoking. PMID- 11474837 TI - Expressing the sense of the Senate to raise the awareness of the devastating impact of diabetes and to support increased funds for diabetes research. PMID- 11474838 TI - Electronic medical records in clinical diabetes care. PMID- 11474839 TI - Zinc and copper in the pathogenesis of amyotrophic lateral sclerosis. AB - 1. Missense mutations in the gene encoding Cu,Zn superoxide dismutase (SOD1) are responsible for causing one form of familial amyotrophic lateral sclerosis (FALS) linked to chromosome 21q. 2. Mutant SOD1-induced disease is clearly related to a toxic gain of function for the abnormal enzyme, and recent work has begun to investigate the mechanisms underlying this toxicity. In addition to its well known and likely beneficial dismutase activity, wild type SOD1 also possesses the ability to participate in other enzymatic reactions that may be injurious to cells including peroxidation or nitration. 3. Many of the SOD1 mutations associated with FALS appear to increase the likelihood that the enzyme will perform either one of these potentially harmful functions resulting in increased hydroxyl radical formation or the addition of nitro groups to tyrosine residues within cellular proteins. PMID- 11474840 TI - Genetic basis of schizophrenia: trinucleotide repeats. An update. AB - 1. Recent developments in technologies permit systematic screening of the entire human genome as a strategy for identification of susceptibility genes of small effect that influence risk to complex traits, like schizophrenia (Schz), inflammatory bowel disease, bipolar affective disorder (BPAD) etc. 2. Schizophrenia is known to have a high heritability and a complex inheritance pattern. Several studies provide evidence that both genes and environment play a role in the etiology of schizophrenia. Linkage studies have observed racial and sex bias in the genetic constitution of schizophrenia. Schizophrenia also manifests clinical anticipation and genomic imprinting. 3. "Dynamic mutations" or "tandem repeat expansions" in DNA, explain a number of observations associated with clinical anticipation and genomic imprinting. In patient populations, the repeat expands well beyond the normal range, altering the biological function of the gene. These sequence are unstable and increase in size between family members in successive generations, giving rise to greater severity of disease. 4. Several workers have reported an association of trinucleotide repeat length with adult- and child-onset schizophrenia. One such expanded allele has been found at the CTG18.1 locus on the 18th chromosome. Other genes known to have similar mutation are SEF2-1, which codes for a helix-loop-helix protein, hSKCa3 gene, which codes for a calcium-activated potassium channel and the transthyretin gene. In schizophrenic patients, significant difference in allele frequency distribution of these genes has been reported. 5. Population based genetic research would not only help identify different subgroups of this of schizophrenia. PMID- 11474841 TI - Prenatal morphine exposure induces age- and sex-dependent changes in seizure susceptibility. AB - 1. Prenatal exposure to morphine induces long-term alterations in seizure susceptibility, which are age-, sex-, and seizure model-specific. 2. Adult male and female rats exposed prenatally to morphine show decreased susceptibility to GABA-regulated seizures. 3. Prenatally morphine-exposed, adult male rats are more sensitive to excitatory amino acid receptor-mediated seizures than control males, control females, or morphine-exposed females. PMID- 11474842 TI - Non-linear dynamic analysis of clozapine-induced electroencephalographic changes in schizophrenic patients--a preliminary study. AB - 1. In order to find the electroencephalographic (EEG) parameters that reflect the effect of clozapine in schizophrenic patients, the authors applied various non linear analyses on multi-channel EEG data drawn from patients before and after a therapeutic trial of clozapine. 2. The correlation dimension was difficult to extract from our limited time series EEG data and the authors did not find a meaningful association with clozapine use. The primary Lyapunov exponent could be reliably calculated but also did not reflect the effect of clozapine. 3. However, the mutual cross-prediction (MCP) algorithm showed potentially meaningful results. The driving system was shifted to the frontal channels after a 4-week trial with clozapine. Moreover, MCP might have a value as a predictor of treatment response. 4. Although preliminary in nature, the MCP might have greater power for interpreting complex changes from channel to channel in EEG induced by clozapine. PMID- 11474843 TI - Deficits of respiratory-cardiac coupling in heavy drinkers. AB - 1. Physiological evidence of chronic alcohol abuse prior to the onset of clinical signs of alcohol dependence is difficult to obtain The purpose of this study was to search for possible non-invasive indicators for chronic alcohol consumption yielding information in addition to conventional biological markers. 2. The authors investigated the relationship between respiratory-cardiac coupling and blood alcohol concentration (BAC) in male subjects who lost their driver's license from drunk driving. 3. We found that subjects who had a high BAC level (0.16-0.31% at the time of offense) show altered respiratory sinus arrhythmia (RSA) and, in particular, an altered heart-rate response to auditory stimulation and compared them to a control group of social drinkers. Normal subjects showed a pronounced acoustic heart-rate response, i.e., particularly during expiration there was a difference between the interbeat-interval (IBI) traces with and without auditory stimulation. Subjects who had lost their driver's license from drunk driving had an overall severely reduced heart-rate response, that was even absent particularly in the subgroup having high BAC values (0.21-0.31%). The authors also found some evidence that in the latter subgroup IBI, RSA, and acoustic heart-rate responses partially recover after a six-month period of abstinence. 4. Specific parameters of the acoustic heart-rate response are changed in our group of alcohol abusers presumably, due to impairment of vagal function. These parameters may therefore be useful to serve as a non-invasive measure of alcohol abuse. PMID- 11474844 TI - Involvement of nuclear factor-kB in the expression of opiate withdrawal. AB - 1. To investigate the role of NF-kB in the expression of opiate withdrawal, the effects of PDTC, an inhibitor of NF-kB activation, was studied on acute opiate withdrawal induced by morphine in vitro. 2. After a 4 min in vitro exposure to morphine, a strong contracture of guinea pig isolated ileum was observed after the addition of naloxone. 3. PDTC (1x10(-8)-5x10(-8)-1x10(-7) M) was able to reduce the naloxone-induced contracture after exposure to the opioid agonist in a concentration-dependent fashion. 4. The results of the present study indicate that NF-kB is involved in the expression of opiate withdrawal thus extending and explaining previous papers performed with dexamethasone and selective arachidonic acid metabolites inhibitors. PMID- 11474845 TI - Modulation of serotonin2A receptor function in rats after repeated treatment with dexamethasone and L-type calcium channel antagonist nimodipine. AB - 1. It has been conceivable that the hypothalamic-pituitary-adrenal (HPA) axis hyperactivity plays an important role in the pathophysiology of depression. In the present study, we have investigated the effect of repeated treatment with dexamethasone on serotonin (5-HT) 1A, 5-HT2A and alpha1-adrenergic receptors in the rat frontal cortex. Moreover, several studies have suggested the effectiveness of L-type calcium channel antagonist nimodipine for the treatment of depression. We also investigated the effect of repeated treatment with nimodipine on 5-HT2A receptor in rats with repeated dexamethasone treatment. 2. Repeated treatment with dexamethasone (1 mg/kg/day for 14 days) increased the density of 5-HT2A receptor, but not 5-HT1A and alpha1-adrenergic receptors in the rat frontal cortex. 3. The density of 5-HT2A receptor in the rat frontal cortex was significantly increased 1 day after repeated treatment with dexamethasone, but was not increased 7 or 14 days after repeated treatment. Wet dog shakes (WDS) induced by (+/-)-1-(4-iodo-2,5-dimethoxyphenyl)-2-aminopropane hydrochloride (DOI), a 5-HT2A receptor agonist, in rats were significantly enhanced 1, 7 and 14 days after repeated treatment with dexamethasone, although the frequency of WDS gradually decreased after repeated treatment. 4. Repeated treatment with nimodipine (5 mg/kg/day for 14 days) attenuated DOI-induced WDS enhanced by repeated treatment with dexamethasone (1 mg/kg/day for 14 days), however, it did not change the density of 5-HT2A receptor. Repeated treatment with dexamethasone decreased locomotor activity and body weight, but repeated treatment with nimodipine did not recover these parameters. 5. The results of the present study suggest that repeated treatment with dexamethasone may selectively increase the 5 HT2A receptor in the rat frontal cortex and affect 5-HT2A receptor-mediated signal transduction. In addition, the intracellular calcium homeostasis by blocking calcium influx through L-type calcium channel may play an important role in the regulation of the 5-HT2A receptor function by dexamethasone. PMID- 11474846 TI - Ethanol enhances naloxone sensitization and disrupts morphine discrimination- comparison to dizocilpine and pentobarbital: explanation of enhancing acute and attenuating chronic effects. AB - 1. Ethanol affects ligand-gated ion channels as a positive modulator of gamma aminobutyric acid (GABA(A)) receptor function and an N-methyl-D-aspartate (NMDA) antagonist. NMDA antagonists attenuate chronic drug effects. Accordingly, we found that ethanol decreased morphine dependence and locomotor sensitization. We now test whether ethanol alters sensitization to the disrupting effects of naloxone on schedule-controlled responding after morphine administration or affects the acute stimulus effects of morphine. 2. Groups of rats, trained to lever-press for food, were co-administered ethanol (1 g/kg; i.p.), the NMDA antagonist dizocilpine (DZ; 0.05 mg/kg; i.p.), the GABA(A) agonist pentobarbital (PB; 3 mg/kg i.p.), or vehicle with morphine (5 mg/kg s.c.). Separate groups received naloxone (0.1-1 mg/kg s.c.) 4-hrs later, prior to food sessions (FR15; 30 min) on three consecutive days. Ethanol enhanced the suppressive effects of higher naloxone doses on all three days. DZ and PB altered this behavior differentially by day and naloxone dose. 3. Next, we examined the effects of ethanol, DZ, PB, and naloxone (0.3 mg/kg; s.c.) on morphine discrimination. Rats, trained to discriminate morphine (3.2 mg/kg s.c.) from saline in a two-lever, food-reinforced procedure, were tested with morphine (0, 1-5.6 mg/kg) after vehicle and drug administrations. Naloxone blocked dose-related responding to morphine, demonstrating pharmacological specificity, and altered response rates. Both ethanol and DZ, but not PB, disrupted morphine-appropriate responding. 4. The paradox that ethanol and DZ attenuate chronic morphine effects while enhancing acute effects may reflect a temporal pattern of primary mu opiate receptor function followed by secondary NMDA-mediated processes induced by morphine administration. PMID- 11474847 TI - Effects of lesioning noradrenergic neurones in the locus coeruleus on conditioned and unconditioned aversive behaviour in the rat. AB - 1. The brain noradrenergic system may have a role in anxiety disorder. This study has examined the effect of bilateral 6-hydroxydopamine lesions of the noradrenergic neurones in the locus coeruleus (LC) of male Lister hooded rats on behaviour produced by unconditioned and conditioned aversive stimuli. 2. The 6 hydroxydopamine (4 microg) lesions markedly reduced the noradrenaline content of the locus coeruleus hypothalamus, frontal cortex and the periaqueductal grey area without altering the levels of either dopamine or 5-hydroxytryptamine measured 14 days after administration. 3. Exposure to ultrasound (20 kHz at 98 dB for 60 sec), an unconditioned aversive stimulus, induced a defence response in the rats characterised by an increase in activity (running and jumping) followed by a period of inactivity (freezing). 4. Lesioning of the LC significantly attenuated the duration of freezing but was without effect on the active phase of the response. A similar reduction in freezing behaviour was seen with LC lesions when rats were exposed (3 hours after the acquisition) to the contextual cue of the conditioned emotion response paradigm. 5. These findings confirm that the locus coeruleus is involved in the regulation of fear-related behaviour in the rat both in an unconditioned and a conditioned model. Furthermore the results indicate that noradrenaline modifies defence behaviour rather than being the principle activating mechanism. PMID- 11474848 TI - Reduction of motor seizures in rats induced by the ethyl bicyclophosphate trimethylolpropane phosphate (TMPP). AB - 1. Trimethylolpropane phosphate (TMPP) is a potent cage convulsant, reported to act through binding to the picrotoxinin and/or benzodiazepine receptor sites of the gamma-aminobutyricA (GABA(A)) ionophore complex. 2. Adult male Fischer-344 rats were pretreated by intraperitoneal (i.p.) injection with either diazepam (DZP) [0.5-5.0 mg/kg], Phenobarbital (PB) [5-20 mg/kg], dizocilpine maleate (MK 801) [0.5-3.0 mg/kg], Tiagabine (TGB) [0.5-5.0 mg/kg], 6,7-dinitro-quinoxaline 2,3-dione (DNQX), [5-20 mg/kg], or scopolamine [SCP] (0.25-1.0 mg/kg) 30 min prior to i.p. injection with a convulsive dose of TMPP (0.6 mg/kg). 3. Rats were rated for occurrence of convulsive activity for 120 min post-injection. Time from TMPP injection to observation of subclinical seizures, generalized (tonic-clonic) seizures, and lethality was rated for each pretreatment group. 4. In general, DZP = PB > TGB in reduction of TMPP subclinical and/or clinical seizures. MK-801, at dose levels inducing near sedation, was also effective in modulation of TMPP induced seizures. SCP or DNQX were generally ineffective in reducing or eliminating TMPP-induced seizures. PMID- 11474849 TI - Killarney 2001: equity, quality, paper clips and staff motivation. PMID- 11474850 TI - Evidence-based practice: does it have a role in paediatrics? PMID- 11474851 TI - Consultant directed or consultant provided hospital services. Achieving an optimal balance. PMID- 11474852 TI - The Forum on Medical Manpower. PMID- 11474853 TI - National guidelines for the active management of HIV in pregnancy. AB - The number of HIV positive women becoming pregnant in Ireland is increasing, with many pregnancies occurring in women from countries of high HIV prevalence. The management of such patients offers the challenge of reducing the risk of maternal fetal transmission of HIV infection, avoiding fetal-infant toxicities, and maintaining future maternal antiretroviral options. Guidelines for the optimal management of HIV disease in pregnancy have been produced by a subgroup of the Irish Infection Society consisting of GenitoUrinary Medicine and Infectious Diseases Consultants. They are based on currently available international data and guidelines. The national guidelines offer a broad management outline for HIV positive pregnant patients. Ultimately, each patient is assessed individually by a multidisciplinary team, and a careful plan for antenatal, intrapartum, and postpartum care is determined. PMID- 11474854 TI - Analysis of outbreaks of infectious intestinal disease in Ireland: 1998 and 1999. AB - Surveillance of general outbreaks of infectious gastroenteritis was introduced in 1998 by the Food Safety Authority of Ireland (FSAI), in co-operation with the eight health boards. A total of 67 general outbreaks of gastroenteritis in Ireland were reported to the FSAI in 1998 and 1999. Over 1900 people were ill as a result of these outbreaks. Four percent required hospitalisation and there were two deaths. The duration of the outbreaks varied between one day and 38 days. Salmonellae (44%) and small round structured viruses (SRSV) (12%) were the most commonly reported pathogens. In 25% of the outbreaks the aetiology was unknown. The commonest settings were restaurants, hotels and take-aways, which accounted for 45% (30/67) of all outbreaks. Sixteen percent of all outbreaks occurred in hospitals and residential institutions. Over half of the outbreaks were reported to be foodborne, 63% of which were due to various serotypes of Salmonella enterica. Eggs were implicated as the vehicle of infection in 13% of all outbreaks. An infected food handler was identified in almost one third of outbreaks, although it could not be established if this had contributed directly to the outbreak. PMID- 11474855 TI - Cost of treating acute myocardial infarction in an Irish teaching hospital. AB - Recent figures indicate that there are approximately 6,500 patients admitted to hospital following acute myocardial infarction (AMI) in Ireland each year. As hospital admission is frequently the most expensive component of healthcare we determined the costs associated with treatment of AMI in a teaching hospital. The costing evaluation was from the hospital perspective and the strategy used was a micro-costing detailed collection of resources used. The average cost of hospital admission for the treatment of AMI was Pound Sterling 3,976. The average cost per day was calculated at Pound Sterling 501. Approximately 50% of hospital costs were associated with ward costs. Procedures accounted for 35% of costs whereas medications contributed just 7% of total costs. PMID- 11474856 TI - A study looking at the effectiveness of developmental screening in identifying learning disabilities in early childhood. AB - This is a retrospective study of children under six years of age referred to the Brothers of Charity Early Intervention Services in County Galway, a service that caters for children under 6 years with learning disabilities. The aim in doing this study was to assess the value of routine developmental screening in identifying children with learning difficulties. This study also investigates the patterns and sources of referral to the remedial services provided by the Brothers of Charity and highlights possible avoidable delays in referral. The results showed that many children were referred for remedial services late. The reasons for late referral included late identification of some children with problems, insufficient co-ordination of community-based services and a lack of awareness of the importance of early intervention in some cases. As some communication disorders such as autism, autistic spectrum disorders and specific language delay may not express themselves until the later part of the second year of life, the 18-24 month developmental assessment is of vital importance. However identification of these disorders can present difficulties and may call for additional training for professionals involved in the developmental screening of children in that age group. The interval between initial identification and referral for remedial care in many cases was more than twelve months. We propose that, in order to minimize this time, children requiring a more in-depth assessment should be assessed by a community-based multidisciplinary team, enabling integrated assessment by the different disciplines and thus speedier referral to remedial services. PMID- 11474857 TI - What happens to stroke patients after hospital discharge? AB - Of 231 stroke patients discharged from hospital, 34 patients (14.7%) had died when reviewed 6 months later. Of 195 survivors, 115 (58%) were independent and living in the community. The remaining 80 (42%) patients were dependent. The majority of dependent patients were in institutional care but 29 (36%) were residing in the community of whom a substantial number were not receiving physiotherapy, occupational therapy or day care. Patients who were dependent in nursing homes were less likely to have received physiotherapy (48% versus 70%) or occupational therapy (28% versus 60%) compared to disabled patients in hospital based extended nursing care. 45 patients (24%) had been re-admitted to hospital although only 48% of patients had been reviewed in hospital outpatients since discharge. 64% of patients were on anti-thrombotic treatment. This survey suggests that 6 months after hospital discharge, most stroke patients are still alive and living in the community. Many of the dependent survivors have ongoing unmet medical and rehabilitation needs. PMID- 11474858 TI - An unusual cause of an acute abdomen. PMID- 11474859 TI - Evaluating cardiac murmurs; are diagnostic tests helpful? AB - The detection of an asymptomatic murmur in a child usually results in a referral to a consultant paediatrician. In most cases a chest radiograph (CXR) and electrocardiograph (ECG) are performed as an aid to diagnosis, however the evidence for this is contradictory. A retrospective chart review of children referred with asymptomatic cardiac murmurs in a one-year period was conducted. We wished to determine whether CXR and ECG are useful diagnostic aids. 81 patient charts were reviewed. Of those patients with a clinical diagnosis of pathological murmur all had normal CXR and ECG. Never the less 2 cases were referred for echocardiogram and were found to have structural heart disease. Of those with a clinical diagnosis of innocent murmur most (81%) had a CXR and ECG and in most cases (96%) these were normal. Those cases with abnormal investigations subsequently were shown to have structurally normal hearts on echocardiogram. We conclude that routine use of CXR and ECG in evaluating asymptomatic cardiac murmurs in children is not useful. PMID- 11474860 TI - A study on the productivity and diseases of camels in eastern Ethiopia. AB - A study concerning performance traits of the Ethiopian camel indicated that, in the camel herds examined, there was one active bull camel for 25 females. The bull camel was 5 years old at puberty; it reached rutting vigour at the age of 9 years, the number of mountings per day was 8 during the breeding season, and the reproduction span was 10 years. The female camel reached puberty at 4 years of age; the age at first calving was 5 years, and the lactation period was one year; the calving interval was 2 years, the calving rate was 50%, and the reproduction span was 10-15 years. The survival rate of the newborn calves was 50%. The average milk yield was 2.5 L per day; the price of camel's milk was higher than that of cow's milk at US$0.5. Adult camels weighed around 500 kg; the dressing out percentage was 52%. Mutton was preferred to camel meat, which came second in popularity, costing US$2/kg. Owing to their poor reproductive performance, camels are not efficient for producing meat. The camels worked for 16 h per day, covering 60 km. Animal health problems encountered were trypanosomosis, camel pox, camel pustular dermatitis, camel cephalopsis, dermatomycosis, mange mite, tick infestation and balantidiosis, most of which mainly affected the young animals. PMID- 11474861 TI - Specific antibodies of Pasteurella multocida in newborn calves of vaccinated dams. AB - Twenty-five newborn Holstein Friesian calves, from dams vaccinated against haemorrhagic septicaemia (HS), were tested repeatedly over the first 6 months of life to monitor the transferred antibody levels against HS. Enzyme-linked immunosorbent assays were used to measure the specific HS antibodies with antigens from Pasteurella multocida strains B:2 and E:2. There was a significant curvilinear relationship between the monitored IgG response and the age of the calves. Peak serum IgG levels were obtained during the period from 8 to 16 weeks of age. Beyond this age, the concentration of IgG in the serum fell away. PMID- 11474862 TI - Detection of anti-listeriolysin O and Listeria monocytogenes in experimentally infected buffaloes (Bubalus bubalis). AB - The kinetics of antibody production against listeriolysin O (ALLO) and the recovery pattern of Listeria monocytogenes from bacteriological samples were studied following oral infection of buffalo calves with 3 x 10(9) cells each of pathogenic L. monocytogenes. Antibodies to LLO appeared by 7-10 days post infection (PI), with a shallow peak between days 16 and 36 PI, when tested by indirect plate-ELISA. The titres of ALLO in all the animals then declined slowly but remained detectable up to day 70 PI. In dot-ELISA, ALLO could be detected by days 5 to 7 PI, and with higher titres than with the plate-ELISA. The pathogen was recovered at low rates as ALLO first appeared but was absent in the faecal, nasal and blood cultures as production of ALLO peaked. PMID- 11474863 TI - A cross-sectional study of bovine tuberculosis in dairy farms in Asmara, Eritrea. AB - Bovine tuberculosis in dairy cattle in Asmara, Eritrea, was studied using a cross sectional study to describe its prevalence and to identify factors associated with it. A total of 72 randomly selected herds were included in the study. The comparative intradermal tuberculin test was used for the diagnosis. Of 1813 individual animals tested, 14.5% were reactors. Thirty herds (41.7%) had at least one reactor but, by defining a reactor herd as any herd with two or more reactors, only 19 (26.4%) herds were classified as reactor herds. Based upon individual animal specificity of 98.5%, the calculated herd specificity was > 99%. A multiple logistic model showed that the presence of exotic breeds was associated with a high risk (odds ratio = 5.70; 95% confidence interval 1.13 28.8). An increased risk was also linked to large herds. Keeping the animals always indoors reduced the risk, but could not be fitted to the model owing to empty cells. PMID- 11474864 TI - Anthelmintic resistance on an organized sheep farm in India. AB - Monitoring anthelmintic resistance in strongyle nematodes by the faecal egg count reduction test and a commercial larval development assay on an organized sheep farm in the semi-arid area of Rajasthan revealed the emergence of resistance to benzimidazoles and rafoxanide and a potential risk of the development of levamisole/tetramisole resistance. A benzimidazole/levamisole combination, avermectins and closantel were each found to be efficacious. PMID- 11474865 TI - Studies on the pathogenicity of Newcastle disease virus isolates in guinea fowl. AB - Newcastle disease viruses isolated from chickens and guinea fowl were characterized as viscerotropic, velogenic strains on the basis of their mean death time, intracerebral pathogenicity index, intravenous pathogenicity index and cloacal and conjunctival mean death time. The pathogenesis of the disease caused by both the strains was studied in 4-week-old guinea fowl. Both strains had an incubation period of 5 days and the birds showed dullness, depression, anorexia, diarrhoea and paralysis of the legs. They also exhibited nervous signs such as incoordination, muscle tremors and trembling of the neck at the advanced stage of the disease. Mortality reached 52% in the group infected with the chicken isolate but it was only 8% in the birds infected with the guinea fowl isolate. No specific changes were observed at post-mortem examination except haemorrhages at the tip of the glands of the proventriculus and in the caecal tonsil. Changes in the lymphoid organs and brain were always present in both the groups. Despite the low mortality, the guinea fowl isolated had multiplied in various organs in the birds. In both groups, the frequency of virus isolation increased from 5 to 10 days post infection. PMID- 11474866 TI - Effect of season on the productivity of camels (Camelus dromedarius and the prevalence of their major parasites in eastern Ethiopia. AB - The productivity and the prevalence rates of the major parasites of camels (Camelus dromedarius) kept under traditional management conditions at Errer valley, Ethiopia were assessed for a year (October 1997-September 1998). The daily milk offtake was significantly higher (p < 0.05) during the wet season (3.12 +/- 0.03 L) than during the dry season (1.49 +/- 0.04 L). Likewise, significantly higher (p < 0.05) daily weight gains (50.68 +/- 0.54 g) were observed during the wet season than during dry season in immature camels aged 1-4 years. Trypanosoma evansi, Sarcoptes scabiei and strongyle parasites were present throughout the year, but their prevalence rates were higher during the rainy months than during the dry months. The minimum and maximum point prevalence rates for T. evansi were 5.4% and 20.6%, respectively. Similarly, the point prevalence rates for S. scabiei also varied from 4.7% during the dry season to 21.7% during the rainy months. The highest strongyle egg counts per gram of faeces and highest point prevalence rate (85.7%) were observed in October, a rainy month, whereas the lowest number of eggs per gram of faeces and the lowest point prevalence rate (61.5%) were recorded in April, a dry month. PMID- 11474867 TI - Growth rate, apparent nutrient digestibility and some blood metabolites of Gwembe valley goats on rations based on crop residues in the hot dry season in Zambia. AB - Twelve Gwembe goats (9 females and 3 males; 12-18-months old; average body weight 18.50 +/- 0.05 kg) were divided into three groups and fed complete rations based on crop residues. Three residues of oilseeds--bambara groundnut shell (BGS), groundnut shell (GNS) and sunflower head (SFH)--were used. The rations were isonitrogenous and isocaloric. The average daily dry matter intake was BGS 0.85 +/- 0.02, GNS 0.88 +/- 0.02 and SFH 0.89 +/- 0.03 kg/head per day, which did not differ from each other significantly (p > 0.05). However, the daily live weight gains were significantly different (p < 0.05). Goats that consumed SFH gained more (84 g; p < 0.05) in live weight than those fed BGS (76 g) or GNS (72 g). Feed conversion followed the pattern of live weight gain. The plasma non-protein N values were 25.1 +/- 0.36, 24.6 +/- 0.38 and 24.9 +/- 0.35 mg/100 ml, while the plasma glucose concentrations were 67.2 +/- 1.44, 65.1 +/- 1.32 and 67.8 +/- 1.49 mg/100 ml, respectively, for goats on BGS, GNS and SFH rations, with no significant difference (p > 0.05) between the three rations. The cost/kilogram for each of the rations were all within the same range, but the cost/kilogram of live weight gain was lower for the SFH ration than for the GNS and BGS rations. The nutrient digestibilities of the crop residues were similar among the treatments. Overall, SFH tended to perform better than GNS or BGS in these complete rations for goats in the hot dry season. PMID- 11474868 TI - Phenotypes including immunocompetence in scavenging local chicken ecotypes in Tanzania. AB - A study was conducted to determine the variations in physical characters and immunocompetence among scavenging local chicken ecotypes in Tanzania. Eighty-four adult scavenging local chickens from four eco-climatic regions of Tanzania were studied. Measurements of adult body weight, body length, shank length and egg weight and observations of plumage colour and pattern, earlobe colour, skin colour and the shape of the comb were conducted. The antibody response to sheep red blood cells, serum haemolytic complement and the cutaneous response to phytohaemagglutinin-P were assessed. Five ecotypes were identified and named Mbeya, Morogoro-medium, Ching'wekwe, Kuchi and Singamagazi. Singamagazi and Kuchi were significantly heavier, with longer shanks and heavier eggs than the other ecotypes. The average adult body weight for males ranged from 1621 g (Mbeya) to 2915 g (Singamagazi). Average female weights ranged from 1108 g (Morogoro-medium) to 2020 g (Singamagazi). Mean egg weights ranged from 37.65 g (Ching'wekwe) to 45.61 (Singamagazi). The Kuchi had mostly rose and walnut combs, while the other ecotypes were mostly single combed. In each ecotype there were chickens with a high or low antibody response to red blood cells, but there was a significant difference between the ecotypes. PMID- 11474869 TI - [Bronchodilator response in elderly patients]. AB - Asthma in the elderly is more severe and a decreased bronchodilating response has been suggested as a contributing factor. There is no agreement on the best way of expressing reversibility. The aim of this study was to evaluate bronchodilator response in elderly patients with asthma with different levels of airway obstruction and expressing reversibility by different indices. A total of 72 asthmatic patients were studied: (FEV1/FVC < 1.64 SEE below predicted). Two groups were considered: Group I: > or = 65 years (71.0 +/- 11.7 years; FEV1 54.0 +/- 16.7% of predicted) and Group II: < 40 years (23.0 +/- 7.7 years, FEV1 67.6 +/- 16.1%). Response to bronchodilators expressed as delta absolute, delta%predicted or delta%maximal was not different between the two groups. Reversibility expressed as delta%initial, however, was lower in younger patients (> 65 years: 22.2 +/- 16.6% vs 40 years: 11.8 +/- 9.9%, p = < 0.005). A covariance analysis was performed using baseline FEV1 as covariate and bronchodilator response was not different between the two groups. Neither delta absolute (r = 0.13, p = NS), delta%predicted (r = 0.06, p = NS) nor delta maximal (r = 0.09, p = NS) showed correlation with age. delta%initial showed weak but significant correlation with age (r = 0.28, p = < 0.05) and marked dependence on baseline FEV1 (r = 0.47, p = < 0.001). Bronchodilator reversibility in the elderly asthmatics is preserved. Expressing reversibility as delta%initial produces differences depending on baseline airway obstruction. PMID- 11474870 TI - [Effect of ipratropium bromide on histamine-induced bronchoconstriction in subjects with chronic airway obstruction]. AB - Ipratropium bromide (IB), a quaternary derivative of atropine has been extensively recommended as the first bronchodilator to be tried in chronic obstructive pulmonary disease (COPD). Despite the large information concerning IB use, a controversy still persists about the lack of non bronchodilator effects (preventive) of inhaled IB. Therefore, the purpose was: to study the effects of IB (80 micrograms) on histamine-induced bronchoconstriction in moderate airway obstruction due to COPD. From outpatient clinic 9 men aged (mean +/- SEM) 57.9 +/ 2.4 yr with smoking history of 54.6 +/- 5.1 pack-yrs and a mean FEV1 = 1.36 +/- 0.08 liters (47.2 +/- 3.8% predicted) were enrolled to participate in this randomized placebo-controlled double blind cross-over study. Each subject attended on 3 occasions (first visit was control day; logPC20 = -0.54 +/- 0.24 mg/ml; geometric mean [MG] = 0.27 mg/ml) for histamine challenge tests using the tidal breathing method after either 4 puffs of IB or placebo aerosol. IB significantly increased baseline FEV1. A correlation between baseline obstruction (FEV1; FEV1/FVC) and bronchodilation with airway hyperreactivity (logPC20) could not be demonstrated. The major finding was that IB attenuated the histamine induced bronchoconstriction (logPC20 = -0.15 +/- 0.17 mg/ml; GM = 0.70 mg/ml) in comparison with placebo (logPC20 = -0.76 +/- 0.22 mg/ml; GM = 0.17 mg/ml; p = 0.018; doubling doses: IB = 2.02 +/- 0.68 vs placebo = -0.62 +/- 0.79; p = 0.024). The lack of correlation between bronchodilator response to IB and the shift in logPC20 might indicate an intrinsic protective role of IB against histamine. Both IB and fenoterol completely resolved the final fall in FEV1 after ending the histamine challenge test. In conclusion, IB diminished histamine induced bronchoconstriction in these subjects with moderate COPD. PMID- 11474871 TI - [Hemorrhagic acute abdomen due to rupture of splenic artery aneurysm. Tomography contribution in the diagnosis]. AB - Three new splenic artery aneurysm rupture cases are presented insisting on the usefulness of abdominal helicoidal computed tomography (AHCT) with intravenous contrast pump. The three patients presented were hospitalized showing acute abdomen associated with non-traumatic hypovolemic shock. In two of the three cases, abdominal computed tomography was used to arrive at the pre-surgical diagnosis. In one of them the abdominal computed tomography with intravenous contrast pump was used, which showed high specificity for the diagnosis, thus becoming a useful methodology whenever arteriography is not available. PMID- 11474872 TI - [Effect of estrogen therapy on the growth of Turner syndrome in girls treated with growth hormone]. AB - Two groups of patients with Turner Syndrome (TS) were studied to analyse the effect of estrogens on their growth. Group 1 (G1): 14 patients treated with growth hormone (GH) who started estrogens (SO) (Premarin) at 15.3 +/- 0.9 years old. Group 2 (G2): 10 girls not treated with GH who started Premarin at 14.3 +/- 2.3 years old. Height SDS improvement in periods of time of GH and GH plus estrogen treatment was evaluated and compared between both groups. The gain in stature during estrogen therapy (final height-projected height at SO) was calculated. In G1 multiple regression was used to examine factors influencing the gain in stature during GH treatment (final height-projected height at the start of GH): bone age at the SO, chronological age at the SO and the time of GH treatment prior to SO. In G1 the height SDS improvement was 0.43 +/- 0.11 cm during GH treatment and 0.59 +/- 0.18 cm during GH plus oestrogens (p = 0.064). The height SDS improvement during estrogen therapy in G2 was 0.14 +/- 0.19 cm, smaller than in G1 (p < 0.001). Gain in stature during estrogen therapy was 5.3 +/- 1.8 cm in G1 and -0.6 +/- 4.2 cm in G2 (p = 0.001). In G1 the time of GH treatment prior to SO was the strongest predictor of the gain in stature during GH treatment (r = 0.89). PMID- 11474873 TI - [Hyperprolactinemia in subclinical hypothyroidism]. AB - Most patients with subclinical hypothyroidism are asymptomatic; however, when this disorder is associated with alterations of the lactotroph, clinical and biochemical manifestations are evident, along with involvement of the gonadotrophic function. In this report, we selected 11 women with hypothyroidism, 5 of them in stage I and 6 in stage II. In all of them, the thyroid function was altered while the most affected was the gonadal axis. The available literature on this subject is scarce, so that we propose a systematic investigation in order to exclude other associated disorders. PMID- 11474874 TI - Immunologic and genetic markers in insulin-dependent diabetes mellitus (type 1) in an Argentine population. AB - The objective was to evaluate the prevalence and association of several markers (islet cell antibodies: ICA, insulin autoantibodies: IAA, glutamic acid decarboxylase antibodies: GADA and ICA512 antibodies: ICA512A) along with HLA DQB1 genotype in type 1 diabetes mellitus of recent onset, including siblings and individuals without any history of this disease, in an Argentine population. A total of 79 children with type 1 diabetes mellitus of recent onset were studied, as well as 79 control children, and 68 healthy siblings of type 1 diabetic cases. IAA, ICA, GADA, ICA512A and HLA DQB1 alleles were determined. Sensitivity was 67.1% for ICA, 36.7% for IAA, 74.6% for GADA and 63.4% for ICA512A. None of the control subjects was positive for the immunological markers. Combined sensitivity of ICA-IAA-GADA was 89.8%, similar to the ICA512A-GADA (87.3%) or ICA512A-GADA IAA combination (91.1%). GADA correlated positively with ICA, but no such correlation was found between IAA, ICA512A and ICA. IAA correlated negatively and GADA positively with age. IAA was associated to DQB1*0201, whereas ICA and ICA512A associated to DQB1*0302. Among siblings, 3/68 (4.4%) were positive for IAA and a single case (1.5%) was positive for GADA and one for ICA512A. Our findings show that the combination of multiple tests increases the sensitivity for prediction, with the ICA512A-GADA combination proving highly sensitive and equivalent to other proposed combinations, such as ICA-IAA-GADA. PMID- 11474875 TI - [Tegumentary leishmaniasis in area with epidemic levels of transmission, Salta, Argentina, 1998]. AB - The incidence of tegumentary leishmaniasis increased in October 1997, in an area close to Oran city, province of Salta. The leishmaniasis' epidemiology and the entomology of this outbreak were studied during June 1998 at Paraje Las Carmelitas and Rio Blanco, places with high reported incidence. The prevalence of infection (Montenegro Skin Test reactive) was 171/1000 inhabitants in Paraje Las Carmelitas and 790/1000 in Rio Blanco, while the rates of leishmaniasis' incidence (active ulcers) were 72/1000 and 790/1000. The prevalence differences among sex and age groups were not significant. The clinical and epidemiologic patterns were consistent with those reported in the area for Leishmania (Viannia) braziliensis. Lutzomyia intermedia was the prevalent species (96%) among 2577 Phlebotominae collected, Lu. shannoni, Lu. cortelezzii y Lu. migonei were also present. The results about distribution of prevalence by age, and spatial distribution of Phlebotominae supported deforestation-residual primary forest transmission hypotheses. Therefore, in Salta the epidemic outbreaks generated by extensive deforestation could be sustained and amplified because the human settlements are located close to the remaining primary forest "spots". Furthermore, in this scenario the risk of human-vector contact increases by the subsistence or recreational activities performed in secondary transitional forests, and the presence of domestic animals around the houses. The results are discussed in the frame of possible control strategies. PMID- 11474876 TI - Human herpesvirus 8 can be transmitted through blood in drug addicts. AB - Human Herpes virus type-8 (HHV-8) seroprevalence was studied in a population of HIV positive intravenous drug users (IVDUs) from Argentina. Analysis of this population also indirectly made it possible to study HHV-8 blood transmission, because these individuals frequently engage in needle sharing behavior and are capable of acquiring a broad array of blood borne pathogens, including Hepatitis B/C virus. The seroprevalence of HHV-8 in IVDUs was compared to a group of non IVDUs and HIV negative individuals. Of the 223 individuals tested, 13.45% were HHV-8 positive, 16.99% in the IVDUs group, and 5.71% in the non-IVDUs. Among HIV positive IVDUs, 25/144 (17.36%) were also HHV-8 seropositive. The seropositivity rate of HHV-8 in HIV negative IVDUs was 11.1%. In contrast, HHV-8 seroprevalence in HIV negative heterosexual individuals without drug usage behavior was even lower (5.71%). The rate of HHV-8 infection in HIV positive IVDUs was three times as high compared to the non IVDU HIV negative individuals, suggesting that IVDU is a risk for HHV-8 infection. Furthermore, it was found that IVDUs showed a very high rate of Hepatitis B/C (52.77%), which also correlate with HHV-8 infection in this population (23.68%). All Hepatitis B/C positive individuals were also HIV positive. Our data confirm other studies showing that individuals who share needles are at risk for acquiring Hepatitis B/C and HIV infections. In addition, our results suggest that they are also at risk to acquiring HHV-8 infection by the same route. PMID- 11474877 TI - Phenotypic and genotypic study of Streptococcus agalactiae in vagina of pregnant women in Argentina. AB - Streptococcus agalactiae (GBS) is a leading cause of serious neonatal infection. In this study we determine the prevalence, serotype distribution and genomic diversity of GBS in vagina of pregnant women. Vaginal swabs of 531 pregnant women were cultured on Columbia Agar Base Blood, GBS Agar Base and Todd Hewitt Broth. GBS were characterized by group and type-specific agglutination. Genomic polymorphism was studied by random amplification of DNA (RAPD). Seventeen patients (3.2%) were positive for GBS, resulting serotype III the most frequent. RAPD detected 16 different RAPD profiles from 21 GBS studied, revealing a good discriminatory power. In this sense, this method showed different genotype from GBS serotype III recovered from successive samples of two patients, suggesting reinfection. In conclusion, the combination of RAPD and serotyping appear promising for epidemiological studies. Finally, findings of reinfection after therapy during pregnancy, led us to suggest performing prenatal GBS screening and intrapartum prophylaxis in order to reduce neonatal risk. PMID- 11474878 TI - [Effects of lysine clonixinate on platelet function. Comparison with other non steroidal anti-inflammatory agents]. AB - One of the mechanisms of action of non steroid antiinflammatory drugs (NSAIDs) consists of inhibition of prostaglandin synthesis. This explains many of the pharmacological effects and adverse events observed in medical practice. Administration of NSAIDs to patients with hemostatic disorders or perioperative conditions entails the risk of bleeding due to inhibition of platelet function. This study deals with platelet changes induced by lysine clonixinate vs diclofenac, ibuprofen and aspirin in classical tests such as platelet count, platelet factor 3 (PF3) activity and platelet aggregation with various inductors and more recent procedures such as P-selectin measurement by flow cytometry. Unlike control drugs, lysine clonixinate did not induce changes in platelet count or function when administered to healthy volunteers at the commonly used therapeutic doses. PMID- 11474879 TI - [Overweight and obesity in children and adolescents of Corrients city. Relationship with cardiovascular risk factors ]. AB - The prevalence of children's obesity is increasing all over the world. It has been related to an increment of all morbimortality causes in the mature age. To determine the prevalence of obesity and other cardiovascular risk factors in adolescence, and correlate them with each other, we designed a cross-sectional study. A survey was applied to 2115 secondary school students, 1212 (57.3%) females and 903 (42.7%) males; blood pressure (BP) and cholesterol level were measured. Mean age: 14.8 +/- 1.6.: Body Mass Index (BMI): 20.6 +/- 3.1, 1838 (86.9%) normal, 230 (10.9%) where overweight and 47 (2.2%) obese, higher in males (OR = 1.46; IC = 1.12-1.91; p = 0.03). Weekly physical activity: 3.9 +/- 2.9 hs. higher in males (p < 0.001). Sedentarism daily hours: 3.2 +/- 1.9. ANTECEDENTS: maternal obesity 282 (13.3%), paternal 311 (14.7%) and in both 116 (6.6%). Regular consumption of fatty foods: 1711 (80.9%). SBP: 107.8 +/- 13.6; DBP: 66.9 +/- 11.5; MBP: 80.5 +/- 10.1. We found hypertension in 64 (3%) prevalent in males (p = 0.002). Cholesterol: 161.9 +/- 29.2; desirable 1283 (60.7%); normal high 628 (29.5%) and high 208 (9.8%). BMI and MBP correlation was: r = 0.25; r2 = 0.06; CI 95%: 0.02-0.11; b coefficient 0.83. We found a positive correlation of BMI with MBP (0.83 mm Hg increment for each point of BMI increment). We found significant association between parental obesity and alcohol consumption and BMI increment. We did not find any association with dietary habits, physical activity or sedentarism. PMID- 11474880 TI - [Printed material distributed by pharmaceutical propaganda agents]. AB - Pharmaceutical sales representatives (drug reps) frequently visit 70% to 90% of physicians during their daily clinical practice and many consider the promotional printed material to be a major source of clinical information. We evaluated samples of the promotional printed material distributed to physicians by drug reps in order to determine whether the data contained in the promotional material is correct and supported by references accessible in Argentina. A consecutive sample of all the promotional material distributed by drug reps in the general internal medicine program (Hospital de Clinicas) was collected between March 15 and April 15, 2000. Reprints and monographs were excluded. Clinical information was reviewed by two general internists and compared to information in a major pharmacology textbook and in an electronic medical information program. References cited were reviewed for correct listing and accessibility in any of the four major medical libraries in Buenos Aires. Of the sixty-four pieces of promotional material collected, thirty were randomly selected and evaluated. In twenty one (70%) the therapeutic effect promoted in advertisement appeared in Goodman & Gilman's 9th edition textbook of pharmacology, in the pharmacology section of the Up-to-Date version 8.1 or in both. Only eighteen (60%) of the thirty promotional printed material evaluated had statements supported by cited references. From a total of 131 references cited in promotional materials, sixty (46%) were incorrectly listed according to the International Committee of Medical Journal Editors. These references were inaccessible. Of the 71 references correctly cited, 49 (69%) were not available in any of the four major medical libraries in Buenos Aires and 8 were available in only two of the libraries. Twenty-two references were reviewed, and in twelve of these (54%), the objective of the research study concurred with the statement of the promotional printed material. Adverse reactions, warnings about drug interactions and contraindications were absent from all promotional printed material. It can be concluded that the promotional printed material distributed by the drug reps in Buenos Aires are biased and provide misinformation more often than not. We recommend that practicing physicians routinely disregard promotional printed material as a source of clinical information. PMID- 11474881 TI - [Thrombosis of great pulmonary vessels causing severe pulmonary hypertension in systemic lupus erythematosus]. AB - We describe a 34 year old male patient with systemic lupus erythematosus (SLE) who developed pulmonary hypertension (PHT) associated with anticardiolipin antibodies. Ten years after diagnosis, an helical CT scan revealed thrombosis of the major pulmonary arteries, which was confirmed by digital pulmonary angiography. An inferior caval vein filter was placed and oral anticoagulation was begun. The patient refused pulmonary thromboendarterectomy and died soon thereafter. Even though PHT is widely described associated with SLE, proximal thrombosis of pulmonary arteries is exceptional. We describe such a patient review the different etiologies, incidence and diagnostic approach of PHT in SLE, focusing on those patients with an effective treatment. PMID- 11474882 TI - [Prostatic adenocarcinoma with neuroendocrine differentiation of the small cell type]. AB - Adenocarcinomas (AC) account for more than 90% of the prostatic malignant neoplasias; a neuroendocrine differentiation (NE) component is to be considered in cases of atypical evolution for its diagnostic and therapeutic relevance. The (NE) originates by proliferation of the NE cells of the acinar epithelium and may develop into two different histologic patterns, a "Carcinoid" type and a "Small Cell" type. We report a patient with a prostatic AC diagnosed 5 years before his death with worsening of the disease together with a return of the prostatic specific antigen (PSA) to normal values and the presence of hepatic metastases of a tumor (NE) of the small cell type. The autopsy confirmed the presence of a prostatic tumor with areas of (AC) type and (NE) small cell type and multiple hepatic metastases (NE). PMID- 11474883 TI - [Vertebral osteoporosis induced by corticoids and cyclosporine ina a patient with Still disease]. AB - We report on a 29-year-old patient who received high doses of prednisone and cyclosporine for the treatment of Still disease. She consulted about dorsolumbar pain leading to physical disability. She presented multiple vertebral fractures, decreased lumbar bone mineral density in the rank of osteoporosis, high bone turnover, and associated hypercalciuria. Cyclosporine and corticoids induced severe changes in bone and mineral metabolism. All patients in treatment with these drugs should undergo radiology, bone densitometry and biochemical determinations of mineral metabolism at the beginning of therapy. Treatment with high doses of intravenous pamidronate (225 mg in 3 months), calcitonin (200/400 IU daily), tiazide (25 mg/daily), and kinesiotherapy mitigated the pain quickly and she recovered motility. We discuss this approach of treating osteoporosis with corticoids and immunosuppressors according to the present knowledge of bone biology. PMID- 11474884 TI - [Telomeres and telomerase activity: their role in aging and in neoplastic development]. AB - Telomeres are specialized structures at the ends of eukaryotic chromosomes, composed of tandem repeats of a repetitive DNA sequence (TTAGGG)n and associated proteins. They have a number of important functions including the protection of chromosomes from end-to-end fusion and degradation. When telomeres become critically short, telomere separation in mitosis cannot be performed properly leading to metaphase telomeric associations (tas) and chromosome instability. This instability can be relevant for neoplastic transformation because it increases the probability of errors that can generate genetic changes critical in the multistep process of transformation, like gene amplification and loss of heterozygosity. The mechanisms involved in tas are unknown, but it could be because of failure in the enzymatic activity of telomerase, a ribonucleoprotein enzyme with an RNA template that directs synthesis of telomeric repeats at chromosome extremities, producing telomeric length stabilization. A progressive telomere shortening with ageing has been shown to occur both in vitro and in vivo. Recent studies have shown an association between the presence of tas and telomeric shortening, and also a correlation between telomere reduction and increased telomerase activity in both solid tumors and hematologic malignancies. The evidence that most human malignancies have telomerase activity would indicate that telomerase could be a prevalent and specific tumor marker, and thus may be a novel and excellent target for anti-cancer therapy. PMID- 11474885 TI - [Cytotoxicity of beta-lapachone, an naphthoquinone with possible therapeutic use]. AB - beta-lapachone (beta-lap) is a lipophilic o-naphthoquinone isolated from the bark of the lapacho tree. Initial observations proved its capability for inhibiting growth of Yoshida tumor and Walker 256 carcinosarcoma. beta-Lap redox-cycling in the presence of reductants and oxygen yields "reactive oxygen species" (ROS: O2-, OH and H2O2) which cytotoxicity led to assume its role in beta-lap activity in cells. beta-Lap inhibited DNA synthesis in Trypanosoma cruzi as well as topoisomerases I and II, poly(ADP-ribose) polymerase (PARP) in different cells. These enzymes are essential for maintaining DNA structure. beta-Lap inhibited growth of a large variety of tumor cells including epidermoid laringeal cancer, prostate, colon, ovary and breast cancer and also different types of leukemia cells. Advances in knowledge of apoptosis ("programmed cell death") and necrosis provided useful information for understanding the mechanism of beta-lap cytotoxicity. Thiol-dependent proteases (Calpaine), kinases (e.g. c-JUN NH2 terminal kinase), caspases and nucleases are involved in beta-lap cytotoxicity. These enzymes activity, as well as ROS production by beta-lap redox-cycling, would be essential for beta-lap cytotoxicity. Diaphorase and NAD(P)H-quinone reductase, which catalyse beta-lap redox-cycling and ROS production, seem to play an essential role in beta-lap activity. On these grounds, clinical applications of beta-lap have been suggested. PMID- 11474886 TI - [Argentine consensus on sleep-related respiratory disorders]. AB - Due to the increased interest of the medical community in sleep disorders an experts meeting was called to establish common criteria for diagnosis, treatment and management of these disorders. Adult prevalence of sleep apnea/hypopnea syndrome (SA/HS) is about 2-4% and increases in the elderly. Snoring and excessive daytime somnolence (EDS) are habitual symptoms. Increased risk to cardiovascular disorders and traffic accidents are the major complications. Increased upper airways resistance syndrome is a recently described syndrome which also involves EDS. A standardized questionnaire was developed and its use was recommended in order to evaluate patients with respiratory sleep disorders (RSD). Polysomnography was established as gold standard in the diagnosis of RSD. Minimal requirement of split night studies and screening studies was also standardized and specific indications were summarized. Medical treatment of obesity in relationship to RSD was analyzed. Nasal continuous positive airways pressure (CPAP) was established as the first choice treatment of SA/HS. Titration of CPAP was standardized. Oral appliances with mandibular advancement could be considered in the treatment of snoring patients without SA/HS and in patients with increased upper airways resistance syndrome. Uvulopalatopharingoplasty can only be performed in snoring patients in whom the presence of SA/HS has been dismissed by polysomnography. Management of patients must include periodic clinical control. EDS must be determined by Epworth test. In order to evaluate CPAP compliance the use of time-controlled devices is highly recommended. PMID- 11474887 TI - [Genetic therapy for the hepatic tumors]. PMID- 11474888 TI - [Serendipity in clinical medicine]. PMID- 11474889 TI - [Diabetes, genetic and immunity]. PMID- 11474891 TI - [Kikuchi-Fujimoto necrotizing lymphadenitis]. PMID- 11474890 TI - [Beta-blockers in the treatment of systolic dysfunction after myocardial infarction. Closing the circle?]. PMID- 11474892 TI - [Conscience objection]. PMID- 11474893 TI - [Serological evidence of cytomegalovirus, hepatitis B and C, Epstein-Barr virus, Toxoplasma gondii, Trypanosoma cruzi and Treponema pallidum in HIV infected patients]. PMID- 11474894 TI - Prevalence of coronary risk factors in healthy adult Kuwaitis. AB - Obesity has been shown to be a serious health risk and is associated with a wide range of comorbid conditions, including diabetes mellitus, coronary heart disease (CHD), gall bladder disease, hypertension, osteoarthritis, and certain cancers. Studies in Kuwait and other Gulf States indicate that obesity is prevalent and may be increasing. Previous studies in Kuwait have mostly focused on the epidemiology of overweight and obesity in children, adolescents, and young adults. Relatively less is known about the prevalence of overweight and obesity and its correlates in older adults for whom obesity may be more closely related to CHD risk. The purpose of this study was to examine the prevalence of overweight and obesity in 9755 adult men and women and to determine the relationship of weight to other risk factors for chronic disease. For the first time we examine a sample of older Kuwaitis to explore the relationship between high BMI, hypercholesterolemia and hyperglycemia. One-third of adult Kuwaitis are obese (BMI > 30 kg/m2), 4.7% of Kuwaiti females have severe obesity (BMI > 40 kg/m2). BMI, cholesterol, and glucose values increase with age across the life span. Older Kuwaitis have a greater constellation of risk factors for chronic diseases. The results of a multiple linear regression model in which cholesterol values were studied, controlling for age, BMI, blood glucose, and sex revealed that age, BMI and sex were significant predictors of cholesterol values. These results indicate that actions to reduce obesity, hypercholesterolemia, and hyperglycemia are needed. PMID- 11474895 TI - Antioxidative properties of water extracts obtained from herbs of the species Lamiaceae. AB - Essential oils and extracts of aromatic herbs obtained by organic solvents have been extensively studied for their antioxidant activity in lipid substrates. Very little is known about the possible presence of antioxidants in polar extracts from herbs used in preparation of infusions and decoctions. In this work water extracts of six different herbs of the Lamiaceae family (dittany, lemon balm, mint, sage, sideritis and sweet marjoram) were prepared. The extracts were examined for their effect against lipid oxidation in comparison to a tea water extract. Sweet marjoram, sage and dittany extracts were found to have a remarkable capacity in retarding lipid oxidation. Examination by thin-layer chromatography of the freeze-dried extracts, before and after hydrolysis, showed that the extracts were rich in bound forms of phenolic compounds such as hydroxycinnamic acids and flavonoids. Rosmarinic and caffeic acids were detected in all extracts with the exception of those from mint and sideritis. These results indicate that certain plants used for the preparation of infusions could be further studied like tea as sources of antioxidants. PMID- 11474896 TI - Studies on germination conditions and antioxidant contents of wheat grain. AB - Germination time and conditions for wheat grain were studied to determine optimum conditions that would maximize the production of antioxidants. Wheat grains were first steeped in water for 24 or 48 h, followed by incubation in the dark for 9 d at 98% RH and 16.5 degrees C. The changes in the concentration of vitamins C and E, beta-carotene, ferulic acid and vanillic acid were monitored over the germination period. Vitamins C and E and beta-carotene were barely detectable in the dry grains. However, upon germination the concentrations of these antioxidant vitamins steadily increased with increasing germination time, reaching their peaks after 7 d at 550 mu/g for vitamin C, 10.92 micrograms/g for alpha tocopherol, and 3.1 micrograms/g for beta-carotene. Concentrations of ferulic and vanillic acids were also increased, reaching their maxima after 7 d at 932.4 micrograms/g and 12.9 micrograms/g, respectively. The grains steeped for 48 h before germination became wet, sticky, yellow-brown color with acidic smell after 7 d. These results suggested that wheat grains steeped for 24 h and germinated for 7 d would produce the most desirable sprouts with respect to antioxidant concentrations and sensory properties. PMID- 11474897 TI - Nutritional composition of uncommon foods and their role in meeting micronutrient needs. AB - Uncommon fruits and vegetables, namely leaves of Bauhenia purpurea Linn., Chenopodium album Linn., Fagopyrum esculentum Moench., and Gleichenia linearis and green and ripe fruits of Ficus roxburghi were analysed for their proximate composition, minerals and vitamin content, in vitro bioavailability of mineral, in vitro protein digestibility and two anti-nutrients, i.e. oxalic and phytic acid. Results showed crude protein, crude fat, minerals, crude fibre, carbohydrate, energy, calcium, iron, and iodine content was in the range of 1.74 to 4.93%, 0.23 to 1.38%, 0.46 to 3.02%, 0.88 to 5.02%, 1.46 to 14.46%, 15 to 76 Kcal, 19 to 355 mg, 1.22 to 6.2 mg and 0.5 to 5.16 mg, respectively, beta carotene, thiamin, riboflavin, niacin and ascorbic acid content ranged between 169 to 3020 micrograms, 0.03 to 0.1 mg, 0.02 to 0.24 mg, 0.07 to 0.87 mg and 3.26 to 173.13 mg per 100 g, respectively. In vitro bioavailability of iron and calcium ranged between 4.62 to 9.23% and 7.30 to 63.48%. In vitro protein digestibility ranged between 9.78 to 14.48%. Findings of the study indicate that all the samples studied are good sources of micronutrients. To provide food security there is need to explore every possible source of nutrients. PMID- 11474898 TI - Underutilised legumes: potential sources for low-cost protein. AB - Seeds of 104 leguminous species belonging to 17 genera were analysed for their protein contents. The promising ones were investigated for fibre, carbohydrate, ash, oil, fatty acids, amino acid profile and trypsin inhibitor activity (TIA). The variation of fibre contents was 4.1-8.9%, carbohydrate 18.4-49.2%, ash 1.8 7.2%, TIA 48.7-87.5 mg/g, oil 1.3-19.8% and protein 11.0-51.6%. The protein content (41-45%) in Acacia mellifera (41.6%), Albizzia lebbek (43.6%), Bauhinia triandra (42.7%), Lathyrus odoratus (42.8%), Parkinsonia aculeata (41.6%), Psophocarpus tetragonolobus (41.9%), Sesbania paludosa (41.2%) and S. sesban (43.8%) was in close proximity to soybean (42.8%), whereas Bauhinia retusa (51.6%), B. variegata (46.5%), Delonix elata (48.7%) and Gliricidia maculata (46.3%) showed higher percentages of protein than soybean. The essential amino acid composition of some of the seed proteins was reasonably well balanced (lysine up to 7.6%). The seeds of Bauhinia retusa (18.6%), B. triandra (16.5%), B. variegata (17.3%), Gliricidia maculata (16.2%), Parkia biglandulosa (18.9%) and Psophocarpus tetragonolobus (19.8%) had a good amount of oil, comparable to soybean (18-22%). The fatty acid composition of some genera/species was quite promising with high amount of unsaturated fatty acids. PMID- 11474899 TI - Functional properties of raw and processed pigeonpea (Cajanus cajan) flour. AB - Emulsification capacity, foam capacity, foam stability, fat and water absorption of raw and processed pigeonpea flour were investigated and compared with those of raw soy flour. The processing methods included soaking in water, soaking and blanching in water and soaking and blanching with trona, a sodium sesquicarbonate salt. Soaking in water was found to reduce the water absorption capacity of pigeonpea flour while soy flour had a slightly higher water absorption capacity than the other processed flours. All the processing treatments increased the fat absorption capacity of pigeonpea flour and the values were also higher than the value obtained for raw soy flour. Data on the emulsifying activities showed that the trona-treated pigeonpea flour had the highest emulsification capacity and was even higher than that of raw soy flour. Soy flour was found to have higher foaming capacity than all the pigeonpea flour samples. PMID- 11474900 TI - Studies on tape ketan--an Indonesian fermented rice food. AB - Preparation of a tape ketan starter material called ragi tape was described by using pure cultures of A. rouxii, S. cerevisiae, P. anomale (HA 122) and P. anomale (HA 893) grown on rice. In a fermentation experiment this self-produced ragi tape was compared with a commercial available ragi tape (NKL). In addition to that, the growth of fungi, yeast and bacteria as well as the biochemical changes during the fermentation process of tape ketan were monitored. To do this, samples were taken periodically after 0, 5, 10, 15, 24, 36, 48 and 60 h fermentation with a commercially available starter (Tebu) at 30 degrees C. Samples were analysed for DM, ash, total N, soluble N, glucose before and after hydrolysis, ethanol, lactic acid, pH and viable cell count. Results show that a palatable product was ready for consumption after 48 h. Finally, tape ketan was prepared using two different samples of Indonesian ragi tape (Tebu) as inoculum and the products were compared by their nutritional constituents. PMID- 11474901 TI - Perceptions of infant cereals and dietary intakes of children aged 4-24 months in a rural South African community. AB - The objectives of this study were to determine (1) mothers' perceptions on infant cereals, and (2) dietary intakes of children aged 4-24 months in a rural South African community. All mothers/caregivers of preschool children who attended community-based growth monitoring posts during January-March 2000 were interviewed. The setting was a low socioeconomic rural African community (Ndunakazi), approximately 60 km northwest of the coastal city of Durban in KwaZulu-Natal, South Africa. Mothers/caretakers (n = 119) of children aged 5 years and under were interviewed to determine perceptions on infant cereals, and to collect retrospective data on complementary feeding practices. Dietary intakes of children aged 4-24 months (n = 50) were determined by 24 h recall. Retrospective data for the 119 children represented in the survey showed that (1) an infant cereal was given as first solid food to 50% of the children; (2) 81% of children had received an infant cereal at some stage, usually at an early age; (3) half of the children had not received any infant cereal after the age of 6 months; (4) the duration for the use of infant cereals was short (on average 3 months); and (5) feeding of an infant cereal was stopped for more than half of the children because the child did not want to eat it any more (disliked it). Most mothers had a positive attitude towards the use of infant cereals, recognizing the health benefits. The cost of the products is high compared to the amount of money spent to feed the household. For children aged 4-24 months of age, the quantities of infant cereal used was inadequate (approximately a quarter of the recommended portion size) and total dietary intakes were below 67% of the recommended levels for calcium, iron, zinc, niacin, vitamin B6 and vitamin B12. It was found that the concept of infant cereals is well accepted and the health benefits thereof are recognized in this rural community. Although these products are available in the area, intakes of children aged 4-24 months were below recommended levels for most of the micronutrients. The availability of culturally acceptable fortified products which are affordable for people in the lower socioeconomic sector is needed. PMID- 11474902 TI - Nutrient intake among female shift workers in a computer factory in Japan. AB - Based on a 4-day questionnaire survey for all meals and snacks consumed by female workers in a computer factory in Japan, consisting of 44 daytime workers and 93 weekly-rotating shift workers (of whom 47 and 46 were engaged in, respectively, early-shift work and late-shift work during the survey week), the present study aimed to clarify the effects of shift work on their nutrient intakes in association with food consumption patterns. Their dietary intakes for 3 working days and an off day were assessed by self-registered food consumption records with the aid of a photographic method, and intakes of energy, protein, fat, carbohydrate, calcium and iron were estimated. The inter-group differences were prominent in the working days. The shift workers, particularly the late-shift workers, took smaller amounts of energy and nutrients than the daytime workers, implying that the former group's nutritional status has been worsened, judged from the recommended dietary allowance for Japanese. Their inadequate nutrient intake was due to lower meal frequency and poor meal quality, both of which were conditioned by shift work. PMID- 11474903 TI - Cadmium in zinc-containing mineral supplements. AB - Seven zinc-containing dietary supplements were analyzed for zinc (Zn) and cadmium (Cd) by inductively coupled plasma/mass spectrometry (ICP/MS). Cadmium was detected in all samples; however, the amount of Cd per 15 mg Zn (the daily US Recommended Dietary Allowance) varied by over 37-fold (0.039 to 1.46 micrograms Cd/15 mg Zn). Supplements with Zn in the form of a gluconate consistently contained the lowest amounts of Cd. Because Cd is a non-essential potentially toxic element for humans, its concentration in nutritional supplements should be minimized and possibly regulated by government-established standards. PMID- 11474904 TI - Perspectives. Dueling managed care bills differ on details. PMID- 11474905 TI - From the Centers for Disease Control and Prevention. Publication of report of tobacco control investment by states. PMID- 11474906 TI - From the Centers for Disease Control and Prevention. HIV incidence among young men who have sex with men--seven US cities, 1994-2000. PMID- 11474907 TI - From the Centers for Disease Control and Prevention. Kernicterus in full-term infants--United States, 1994-1998. PMID- 11474908 TI - Pacifiers and breastfeeding. PMID- 11474909 TI - Defining and diagnosing irritable bowel syndrome. AB - Approximately 20% of the general population has irritable bowel syndrome (IBS). Although the majority of these individuals do not consult a physician, IBS accounts for 25% of visits to a gastroenterologist and up to 12% of visits to a primary care physician. Consequently, the direct and indirect costs associated with IBS are estimated at $8 billion annually. IBS symptoms, with no apparent structural pathology, include altered bowel habits, abdominal pain/discomfort, and bloating. The Rome II criteria, a standardized guideline for the diagnosis of IBS, contains in its definition abdominal pain or discomfort associated with altered bowel habits. Bloating may often be present. Three patient subgroups are defined according to the predominant bowel symptom: constipation, diarrhea, or alternating constipation and diarrhea. Hematology, fecal occult blood test, flexible sigmoidoscopy, and lactose intolerance evaluations are recommended for all patients demonstrating symptoms of IBS. When indicated, tests are recommended to rule out bacterial or parasitic infections, pelvic floor muscle dyssynergia, colonic inertia, peptic ulcer, or inflammatory bowel disease. PMID- 11474910 TI - The role of serotonin in the pathophysiology of irritable bowel syndrome. AB - Coordinated activities of the central, autonomic, and enteric nervous systems modulate intestinal motor, sensory, and secretory activities that may contribute to the triad of dysfunction (altered motility, altered sensation, and psychosocial distress) observed in patients with irritable bowel syndrome (IBS). Autonomic modulation of gastrointestinal (GI) function occurs via the actions of neurotransmitters and neuromodulators such as serotonin (5-hydroxytryptamine, or 5-HT), norepinephrine, and dopamine. Of those modulators, serotonin has received the most attention with respect to disorders of GI function. Serotonin exerts its effects via neurocrine, paracrine, and endocrine pathways. Recent studies have demonstrated that serotonin, acting primarily through 5-HT3 and 5-HT4 receptors, is intricately involved in initiating the peristaltic reflex and facilitating intraluminal secretions. Serotonin receptors mediate reflex control of GI motility and secretion and may influence the perception of bowel function and pain under some circumstances. GI motor activity and sensory dysfunction in patients with IBS may be a result of alterations in serotonin levels or associated 5-HT receptors. Serotonin agonists and antagonists such as tegaserod, a 5-HT4 agonist, may offer new treatments that normalize GI motor and sensory functions in patients with disorders of GI function. PMID- 11474911 TI - Drug therapy options for patients with irritable bowel syndrome. AB - Existing pharmacotherapeutic options for the treatment of patients with irritable bowel syndrome (IBS) are limited in treating the multiple symptoms associated with the disorder. There is much interest in the use of serotonin agents as new therapeutics. Acting primarily through 5-HT3 and 5-HT4 receptors, serotonin elicits changes in motor function and possibly visceral sensation. Two serotonin agents were developed specifically for IBS: tegaserod, a 5-HT4 receptor partial agonist, and alosetron, a 5-HT3 receptor antagonist (which is no longer available). Phase III clinical trial data show that during a 12-week treatment period with tegaserod, IBS patients with abdominal pain and discomfort, bloating, and constipation experienced significant global relief (i.e., improvement in overall well-being, abdominal pain, and bowel habit) compared with placebo. Improvement in bowel movement frequency and consistency was achieved and pain was relieved by 1 week. During 12 weeks of treatment, alosetron was shown to elicit significant relief of abdominal pain and discomfort compared with placebo or mebeverine in female IBS patients with diarrhea. Alosetron slowed colonic transit and treatment efficacy was apparent after a week of treatment. Another 5-HT4 receptor agonist, prucalopride, which is being developed for chronic constipation, accelerates colonic transit and increases stool frequency. Therefore, this agent may be of benefit in IBS patients with constipation. PMID- 11474912 TI - Irritable bowel syndrome: toward a cost-effective management approach. AB - OBJECTIVE: To examine the economic implications of current irritable bowel syndrome (IBS) management practices and formulate recommendations based on these implications. METHODS: Relevant English-language research publications in which the direct and indirect costs of IBS were examined, identified using a search of records contained in Medline. RESULTS: Review of the identified publications indicates that in Western nations, IBS management is associated with high direct costs (particularly for diagnostic testing, office visits, pharmacotherapy, and emergency department visits). Indirect costs, associated with lost wages and decreased productivity, account for the largest proportion of the IBS economic burden. Moreover, rapid projected growth in IBS disease-related costs indicates a need for more focused attention toward improved treatment of IBS. More cost effective management might be achieved by diagnosing and instituting nonpharmacologic and pharmacologic management earlier in the disease process. Under such an approach, patients are classified based on symptoms and a therapeutic trial is begun. More extensive, expensive diagnostic testing is reserved for patients refractory to treatment or for whom serious disease must be ruled out. CONCLUSION: IBS is a condition with high direct and indirect costs. Management strategies should be evaluated both on their clinical efficacy and on their cost effectiveness. As new, IBS-specific pharmacotherapies become available, the ability to diagnose and manage the condition in a cost-effective manner can be improved. PMID- 11474913 TI - Community-oriented oral health promotion for infants in Jerusalem: evaluation of a program trial. AB - OBJECTIVES: This study sought to measure the effect of a community health education program on reported infants' bottle-feeding practices and infants' toothbrushing behavior, with or without distribution of toothpaste and toothbrushes. METHODS: In this quasi-experimental comparison group design study conducted in mother and child health centers in Jerusalem, parents of 727 children were surveyed by telephone at baseline and six months later. The cohort of infants was aged 6-12 months at baseline. The program group received structured health education. The control group received no organized educational intervention. Within the program and control groups, half of the centers were randomly given toothpaste and toothbrushes. RESULTS: Parents' reports revealed a secular 32.5 percent increase in toothbrushing for infants with no intervention, 45.1 percent for infants only receiving toothpaste and toothbrushes, 43.7 percent for infants only receiving the health education program, and a 60.4 percent increase for infants receiving health education together with toothpaste and toothbrushes (chi-square, P = .0002). Modification of bottle-drinking practices, in this program, was unsuccessful. CONCLUSION: The free distribution of toothpaste and toothbrushes, together with an oral health education program, is recommended as a potentially practical and effective method of promoting early oral hygiene practices. PMID- 11474914 TI - Training status and interest in certification of nondiplomate faculty teaching predoctoral dental public health. AB - OBJECTIVES: A survey was conducted to better understand the training needs of faculty members without dental public health (DPH) specialty board certification who teach DPH to dental students. METHODS: An 11-item questionnaire was sent to 193 non-DPH diplomate faculty members at US dental schools who were dentists and at least one of the following: a member of the American Association of Dental Schools Community and Preventive Dentistry Section, a referral from an academic American Board of Dental Public Health diplomate, a DPH faculty listed on the school's Web pages, a DPH contact from the AADS Institutional Directory, or the school's dean if no other contact. RESULTS: A 70 percent response rate was obtained. Seventy-nine percent of the respondents taught at least one national board-related DPH topic. Among these faculty members, 67 percent have or are in training for the master of public health, 26 percent have completed or are in a DPH residency, and 63 percent desire training in one or more of the DPH topics. The majority (64%) does not plan to take the specialty exam, while 28 percent plan to take the exam within five years. About half reported no personal incentives to take the exam and 39 percent perceived no institutional incentives. CONCLUSIONS: These nondiplomate teachers of predoctoral DPH desire training, but appear to have barriers and perceive few benefits to achieving DPH board certification. PMID- 11474915 TI - A two-year longitudinal study of dental caries in permanent first molars of Korean elementary schoolchildren. AB - OBJECTIVE: This longitudinal study investigated dental caries increment in permanent first molars of Korean elementary schoolchildren. METHODS: A convenience sample of 722 children aged 7-9 years attending one urban elementary school was examined at baseline, with follow-up examinations at one and two years. Coronal surfaces of permanent first molars were scored with regard to caries experience and sealant status. RESULTS: Among sound occlusal surfaces at baseline, 21 percent of upper and 25 percent of lower surfaces developed caries during the two-year interval. In teeth that erupted between baseline and the first follow-up exam, over 10 percent of occlusal surfaces developed caries. Pit and fissure caries accounted for 93 percent of all new carious surfaces, while sealants had been placed on 16 percent of occlusal surfaces during the study. CONCLUSIONS: Recognizing the limitations of this convenience sample, dental sealants should be used more widely in this Korean population, and should be applied soon after tooth eruption. PMID- 11474916 TI - Response to the JPHD's guest editorial: qualitative research--does it have a place in dental public health? PMID- 11474917 TI - Patterns of fluoride intake from birth to 36 months. AB - OBJECTIVES: Dental fluorosis prevalence has increased in the United States, Canada, and other nations due to the widespread availability of fluoride in many forms, with fluoride ingestion during the first three years of life appearing most critical in fluorosis etiology. With few contemporary studies of fluoride ingestion in this age group, the purpose of this paper is to describe patterns of estimated fluoride ingestion from birth to 36 months of age from water, dentifrice, and dietary fluoride supplements and combined. METHODS: Repeated responses to separate series of questions about water intake, use of fluoride dentifrice, and use of fluoride supplements were collected by questionnaire as part of the longitudinal Iowa Fluoride Study and used to estimate fluoride intake. Estimated intake is reported by source and combined at different ages. Effects of subject age and other covariates on fluoride intake were assessed using regression methods appropriate for the analysis of correlated data. RESULTS: For most children, water fluoride intake was the predominant source, especially through age 12 months. Combined daily fluoride intake increased through 9 months, was lower at 12 and 16 months, and increased again thereafter. Mean intake per unit body weight (bw) was about 0.075 mg F/kg bw through 3 months of age, 0.06 mg F/kg bw at 6 and 9 months, 0.035 mg F/kg bw at 12 and 16 months, and 0.043 mg F/kg bw from 20-36 months. Depending on the threshold chosen (e.g., 0.05 or 0.07 mg F/kg bw), variable percentages of the children exceeded the levels, with percentages greatest during the first 9 months. Regression analyses showed fluoride intake (mg F/kg bw) from 1.5-9 months to decrease with increasing child's age, mother's age, and mother's education, with a complex three-way interaction among these factors. From 12-20 months, fluoride intake increased with increasing child age and decreased with increasing mother's age. No statistically significant relationships were found for fluoride intake from 24-36 months. CONCLUSIONS: There is considerable variation in fluoride intake across ages and among individuals. Longitudinal studies may be necessary to fully understand the relationships between fluoride ingestion over time and development of fluorosis. PMID- 11474918 TI - An economic evaluation of community water fluoridation. AB - OBJECTIVE: The purpose of this research was to assess the local cost savings resulting from community water fluoridation, given current exposure levels to other fluoride sources. METHODS: Adopting a societal perspective and using a discount rate of 4 percent, we compared the annual per person cost of fluoridation with the cost of averted disease and productivity losses. The latter was the product of annual dental caries increment in nonfluoridated communities, fluoridation effectiveness, and the discounted lifetime cost of treating a carious tooth surface. We obtained or imputed all parameters from published studies and national surveys. We conducted one-way and three-way sensitivity analyses. RESULTS: With base-case assumptions, the annual per person cost savings resulting from fluoridation ranged from $15.95 in very small communities to $18.62 in large communities. Fluoridation was still cost saving for communities of any size if we allowed increment, effectiveness, or the discount rate to take on their worst-case values, individually. For simultaneous variation of variables, fluoridation was cost saving for all but very small communities. There, fluoridation was cost saving if the reduction in carious surfaces attributable to one year of fluoridation was at least 0.046. CONCLUSION: On the basis of the most current data available on the effectiveness and cost of fluoridation, caries increment, and the cost and longevity of dental restorations, we find that water fluoridation offers significant cost savings. PMID- 11474919 TI - Prevalence of dental fluorosis in the primary dentition. AB - OBJECTIVES: This paper presents data on the prevalence of primary tooth fluorosis among children residing in Iowa, and the relationships between fluorosis prevalence and selected measures of fluoride exposures. METHODS: Children in the study cohort were followed prospectively during the first year of life. This study assessed their home water fluoride concentrations and use of fluoride dentifrice or dietary fluoride supplements. A total of 637 children (320 females and 317 males) were examined for fluorosis using a modification of the TSIF index at age 4 1/2 to 5 years, with 90.4 percent having intact primary dentitions. RESULTS: 74 children (11.6%) had fluorosis present on one or more of their primary teeth, and 71 children (11.1%) had two or more teeth affected. Nearly all fluorosis was mild, with the primary second molar teeth most commonly affected. Fluorosis was significantly associated with higher water fluoride concentration, but not with the use of dentifrice or fluoride supplements. CONCLUSIONS: The results of this study show that primary tooth fluorosis is relatively uncommon, but is most frequently seen on the posterior teeth, particularly the primary second molars, which form at later stages of development. This finding suggests that primary tooth fluorosis is mostly a postnatal phenomenon, and is associated with higher water fluoride levels. PMID- 11474920 TI - US drinking water: fluoridation knowledge level of water plant operators. AB - OBJECTIVES: We determined the knowledge level of water plant operators who fluoridate drinking water, and we compared small and large water plants. METHODS: A pretested survey was sent to 2,381 water plant operators in 12 states that adjust the fluoride concentration of drinking water. A z-test for proportion was used to test for statistical difference between small and large plants at alpha = 0.05. Small water plants were those treating less than 1 million gallons of water daily. RESULTS: Eight hundred small and 480 large water plant operators responded, resulting in a response rate of 54 percent. Two-thirds of water plant operators correctly identified the optimal fluoride level, but more than 20 percent used a poor source for choosing the optimal level. Only one-fourth of operators were able to maintain the fluoride concentration to within 0.1 mg/L of the optimal concentration. A significantly greater proportion of operators at large water plants than at small water plants reported that they were able to maintain a fluoride concentration to within 0.1 mg/L of the optimal concentration (33.5% vs 21.3%, z = 4.74, P < .05). CONCLUSIONS: Although most operators correctly identified the optimal fluoride level, small water plant operators were less likely to use accurate reasoning for choosing that level and in maintaining fluoride concentrations within 0.1 mg/L of that level than large water plant operators. PMID- 11474921 TI - Fluid consumption related to climate among children in the United States. AB - OBJECTIVE: Recommended fluoride concentrations in US public water systems are between 0.7-1.2 ppm, depending on the mean daily maximum temperature. This range assumes that water intake is higher in warmer than in cooler climates, based on research from the 1950s. The aim of this analysis is to relate fluid consumption among American children aged 1-10 years to the local climate under modern conditions. METHODS: The quantities of daily total fluid intake per body weight (ml/kg) and plain water intake per body weight (ml/kg) of children were calculated from the 24-hour recall diet survey in the third National Health and Nutrition Examination Survey (NHANES III, 1988-94). The mean daily maximum temperature from 1961 to 1990, averaged for the month during which the NHANES III exam was conducted, was obtained for each survey location from the US Local Climate Historical Database. Multiple regression analysis was conducted using SAS and SUDAAN. RESULTS: Fluid intake was significantly associated with age, sex, socioeconomic status (SES), and race and ethnicity. No significant association could be found between the amount of either total fluid or plain water intake and mean daily maximum temperature, either before and after controlling for sex, age, SES, and race or ethnicity. CONCLUSIONS: Results indicate that there is no evidence that fluid consumption among children is significantly related to mean temperature in modern conditions. This suggests that the national temperature related guidelines for fluoride concentration in drinking water may be due for reevaluation. PMID- 11474922 TI - Simultaneous MEG and EEG source analysis. AB - A method is described to derive source and conductivity estimates in a simultaneous MEG and EEG source analysis. In addition the covariance matrix of the estimates is derived. Simulation studies with a concentric spheres model and a more realistic boundary element model indicate that this method has several advantages, even if only a few EEG sensors are added to a MEG configuration. First, a simultaneous analysis profits from the 'preferred' location directions of MEG and EEG. Second, deep sources can be estimated quite accurately, which is an advantage compared to MEG. Third, superficial sources profit from accurate MEG location and from accurate EEG moment. Fourth, the radial source component can be estimated, which is an advantage compared to MEG. Fifth, the conductivities can be estimated. It is shown that conductivity estimation gives a substantial increase in precision, even if the conductivities are not identified appropriately. An illustrative analysis of empirical data supports these findings. PMID- 11474923 TI - Numerical evaluation of shaped surface coil sensitivity at 63 MHz. AB - Surface coils are widely used in magnetic resonance studies due to their superior signal to noise (SNR) properties. When shaping planar coils to cylindrical surfaces, the region with maximum sensitivity migrates from the coil plane towards the centre of the shaping radius. The influence of the coil current, the probe and tissue dimensions, the electrical tissue properties and the operating frequency on the B1 field strength of a coil has been studied using statistical methods. This analysis allows the dependence of the axial SNR distribution of circular and square surface coils on shaping radius and coil dimensions to be evaluated quantitatively using 3D finite element methods. An empirically derived equation describing the dependence of the SNR distributions on coil geometry and depth allows the optimum coil dimensions to be predicted for a given shaping radius and desired optimized depth of sensitivity. Simulations are validated experimentally using both B1 and SNR mapping techniques. A comparison between the axial SNR of circular and square coils demonstrated equal SNR distributions of coils with equivalent area at depth. PMID- 11474924 TI - A spatially varying compton scatter correction for SPECT utilizing the integral Klein-Nishina cross section. AB - An algorithm correcting for the fraction of scattered events in SPECT and planar imaging is proposed. The algorithm utilizes a pixel-based multi-channel analyser for data acquisition. The method was designed to operate on a local level by three subtraction steps: (a) Subtracting a modified Klein-Nishina single scatter distribution, pixel by pixel, from the events obtained experimentally in the upper half of the photo-peak window. (b) Subtracting a mirrored distribution of the unscattered events hence obtained from that of total events in the lower half of the window, thus giving the scatter distribution in this part of the window. (c) Subtracting the sum of the scatter distributions in both window halves from the corresponding sum of total events in order to obtain the unscattered photons within the photo-peak window. The accuracy of the method was validated experimentally, using a new rCBF phantom allowing for imaging in matter corresponding to soft tissue and approximately in air, respectively. After correction for photon scattering and attenuation, the regional difference in SPECT values in soft tissue equivalent matter and in low-density matter (simulating air) was only 1.5 +/- 7.2% (mean +/- 1 SD), thus indicating a high accuracy of the correction method. Provided that an accurate and stable pixel peak-alignment routine is available, the method can be applied using a minimum of three windows. PMID- 11474925 TI - Comparison of modelled and observed in vivo temperature elevations induced by focused ultrasound: implications for treatment planning. AB - Two numerical models for predicting the temperature elevations resulting from focused ultrasound heating of muscle tissue were tested against experimental data. Both models use the Rayleigh-Sommerfeld integral to calculate the pressure field from a source distribution. The first method assumes a source distribution derived from a uniformly radiating transducer whereas the second uses a source distribution obtained by numerically projecting pressure field measurements from an area near the focus backward toward the transducer surface. Both of these calculated ultrasound fields were used as heat sources in the bioheat equation to calculate the temperature elevation in vivo. Experimental results were obtained from in vivo rabbit experiments using eight-element sector-vortex transducers at 1.61 and 1.7 MHz and noninvasive temperature mapping with MRI. Results showed that the uniformly radiating transducer model over-predicted the peak temperature by a factor ranging from 1.4 to 2.8, depending on the operating mode. Simulations run using the back-projected sources were much closer to experimental values, ranging from 1.0 to 1.7 times the experimental results, again varying with mode. Thus, a significant improvement in the treatment planning can be obtained by using actual measured ultrasound field distributions in combination with backward projection. PMID- 11474926 TI - Simultaneous optimization of cryoprobe placement and thermal protocol for cryosurgery. AB - We demonstrate that it is possible to simultaneously optimize multiple cryoprobe placements and their thermal protocol for one freeze-thaw cycle. A numerical optimization algorithm is used and three different forms of objective function are examined in terms of algorithm convergence rate, minimum value of the chosen objective function, temperature-volume histograms and isotherm distributions. The optimization results depend on the initial values of the variables, the form of the objective function, optimization goals and the mathematical method adopted for gradient calculation. The proposed optimization model offers significant advantages over the previously reported semi-empirical approach to conformal cryotherapy, such as the ability to handle an unlimited number of variables and eliminating the need for the user input between iterations, thereby reducing, if not removing, the subjectivity of cryosurgery treatment planning. PMID- 11474927 TI - Series model volume effects in a population of non-identical patients: how low is low? AB - Working with several mechanisms of critical local tissue damage, formulae are analytically derived that describe normal tissue complication probabilities (ntcps) for series-type radiotherapy complications arising in heterogeneous patient populations. Using the formulae, values are calculated for deltaD50(10) the increase in dose leading to a 50% series-type complication rate (D50) when irradiated organ volume is reduced tenfold. From the structure of the ntcp formulae derived, it follows that dose-levels leading to clinically relevant serious complication rates (less than 5%) will change less with irradiated volume than will D50. Calculated values of deltaD50(10) for the heterogeneous series model are low-generally less than 6 Gy; such values are much lower than those calculated for the non-heterogeneous series model (27-37 Gy). These results suggest that if the dose-limiting toxicity of a radiotherapy treatment is a series-type complication with a local damage mechanism similar to any of those studied in this work, then even very substantial improvements in technique leading to large reductions in highly dosed normal tissue volumes-would be unlikely to allow a useful degree of escalation of the dose delivered to the tumour, unless highly dosed normal tissue volumes can be reduced below the length scale of a functional subunit. PMID- 11474928 TI - Ordered subset reconstruction for x-ray CT. AB - Statistical methods for image reconstruction such as the maximum likelihood expectation maximization are more robust and flexible than analytical inversion methods and allow for accurate modelling of the counting statistics and photon transport during acquisition of projection data. Statistical reconstruction is prohibitively slow when applied to clinical x-ray CT due to the large data sets and the high number of iterations required for reconstructing high-resolution images. Recently, however, powerful methods for accelerating statistical reconstruction have been proposed which, instead of accessing all projections simultaneously for updating an image estimate, are based on accessing a subset of projections at the time during iterative reconstruction. In this paper we study images generated by the convex algorithm accelerated by the use of ordered subsets (the OS convex algorithm (OSC)) for data sets with sizes, noise levels and spatial resolution representative of x-ray CT imaging. It is only in the case of extremely high acceleration factors (higher than 50, corresponding to fewer than 20 projections per subset), that areas with incorrect grey values appear in the reconstructed images, and that image noise increases compared with the standard convex algorithm. These image degradations can be adequately corrected for by running the final iteration of OSC with a reduced number of subsets. Even by applying such a relatively slow final iteration, OSC produces almost an equal resolution and lesion contrast as the standard convex algorithm, but more than two orders of magnitude faster. PMID- 11474929 TI - Performance evaluation of the microPET P4: a PET system dedicated to animal imaging. AB - The microPET Primate 4-ring system (P4) is an animal PET tomograph with a 7.8 cm axial extent, a 19 cm diameter transaxial field of view (FOV) and a 22 cm animal port. The system is composed of 168 detector modules, each with an 8 x 8 array of 2.2 x 2.2 x 10 mm3 lutetium oxyorthosilicate crystals, arranged as 32 crystal rings 26 cm in diameter. The detector crystals are coupled to a Hamamatsu R5900 C8 PS-PMT via a 10 cm long optical fibre bundle. The detectors have a timing resolution of 3.2 ns, an average energy resolution of 26%, and an average intrinsic spatial resolution of 1.75 mm. The system operates in 3D mode without inter-plane septa, acquiring data in list mode. The reconstructed image spatial resolution ranges from 1.8 mm at the centre to 3 mm at 4 cm radial offset. The tomograph has a peak system sensitivity of 2.25% at the centre of the FOV with a 250-750 keV energy window. The noise equivalent count rate peaks at 100-290 kcps for representative object sizes. Images from two phantoms and three different types of laboratory animal demonstrate the advantage of the P4 system over the original prototype microPET. including its threefold improvement in sensitivity and a large axial FOV sufficient to image an entire mouse in a single bed position. PMID- 11474930 TI - Temporal backward projection of optoacoustic pressure transients using fourier transform methods. AB - In medical imaging different techniques have been developed to gain information from inside a tissue. Optoacoustics is a method to generate tomography pictures of tissue using Q-switched laser pulses. Due to thermal and pressure confinement, a short light pulse generates a pressure distribution inside tissue, which mirrors absorbing structures and can be measured outside the tissue. Using a temporal back-projection method, the pressure distribution measured on the tissue surface allows us to gain a tomography picture of the absorbing structures inside tissue. This study presents a novel computational algorithm, which, at least in principle, yields an exact reconstruction of the absorbing structures in three dimensional space inside the tissue. The reconstruction is based on 2D pressure distributions captured outside at different delay times. The algorithm is tested in a simulation and back-projection of pressure transients of a small absorber and a single point source. PMID- 11474931 TI - Wedge-shaped applicator for additional light delivery and dosimetry in the diaphragmal sinus during photodynamic therapy for malignant pleural mesothelioma. AB - In situ light dosimetry during photodynamic therapy (PDT) of malignant pleural mesothelioma (MPM) after tumour resection facilitates the delivery of a controlled light distribution to the inner thoracic surface. Illumination of the diaphragm-induced sinus, however, remains difficult. Our aim was to develop a wedge-shaped light applicator with incorporated light dosimetry to deliver an additional fluence limited to the sinus. The wedge-shaped applicator contains a cylindrical diffuser for light delivery and two isotropic detectors for simultaneous light dosimetry. These detectors were placed at strategic positions where the fluence rate is maximal or minimal (middle and edge). Prior to its clinical use, the performance of the sinus light applicator was tested in several optical tissue phantoms with different optical properties. The fluence rate distribution over the surface of the applicator showed little change when the wedge was submerged in four different optical phantoms. During clinical PDT of MPM the applicator had to be re-located manually four times in order to give an additional fluence of approximately 2 J cm(-2) to the entire sinus. The light applicator enables dosimetry-controlled light delivery for additional illumination of the sinus region that is often under-illuminated during thoracic integral illumination of MPM. PMID- 11474932 TI - Monitoring of deep brain temperature in infants using multi-frequency microwave radiometry and thermal modelling. AB - In this study we present a design for a multi-frequency microwave radiometer aimed at prolonged monitoring of deep brain temperature in newborn infants and suitable for use during hypothermic neural rescue therapy. We identify appropriate hardware to measure brightness temperature and evaluate the accuracy of the measurements. We describe a method to estimate the tissue temperature distribution from measured brightness temperatures which uses the results of numerical simulations of the tissue temperature as well as the propagation of the microwaves in a realistic detailed three-dimensional infant head model. The temperature retrieval method is then used to evaluate how the statistical fluctuations in the measured brightness temperatures limit the confidence interval for the estimated temperature: for an 18 degrees C temperature differential between cooled surface and deep brain we found a standard error in the estimated central brain temperature of 0.75 degrees C. Evaluation of the systematic errors arising from inaccuracies in model parameters showed that realistic deviations in tissue parameters have little impact compared to uncertainty in the thickness of the bolus between the receiving antenna and the infant's head or in the skull thickness. This highlights the need to pay particular attention to these latter parameters in future practical implementation of the technique. PMID- 11474933 TI - Helical antenna arrays for interstitial microwave thermal therapy for prostate cancer: tissue phantom testing and simulations for treatment. AB - Interstitial microwave therapy is an experimental treatment for prostate cancer. The objective of this work was to measure the power deposition (specific absorption rate, SAR) patterns of helical microwave antennae both individually and in array patterns that would be useful for clinical treatment protocols. Commercial helical antenna 3D SAR patterns were measured in muscle equivalent phantoms using a thermographic technique. Two array patterns were tested: a 'square' and a 'crescent' array, both surrounding the urethra. To assess the feasibility of pre-treatment planning, the measured SAR patterns were input to a treatment planning computer simulation program based on a series of trans-rectal ultrasound images from a prostate cancer patient. The simulation solved the Pennes linear bioheat heat transfer equation in prostate tissue, with the aim of achieving a target of 55 degrees C at the prostate periphery while not allowing normal surrounding tissues (bladder, urethra, rectum) to rise above 42 C. These criteria could not be met with the square array but they could be met with the crescent array, provided that the prostate was first dissected away from the rectum. This can be done with a procedure such as 'hydrodissection', where sterile saline is injected to separate the prostate and rectum. The results of these SAR measurements and heat transfer simulations indicate that arrays of helical antennae could be used for safe and effective thermal therapy for prostate cancer. PMID- 11474934 TI - Regional hyperthermia applicator design using FDTD modelling. AB - Recently published results confirm the positive effect of regional hyperthermia combined with external radiotherapy on pelvic tumours. Several studies have been published on the improvement of RF annular array applicator systems with dipoles and a closed water bolus. This study investigates the performance of a next generation applicator system for regional hyperthermia with a multi-ring annular array of antennas and an open water bolus. A cavity slot antenna is introduced to enhance the directivity and reduce mutual coupling between the antennas. Several design parameters, i.e. dimensions, number of antennas and operating frequency, have been evaluated using several patient models. Performance indices have been defined to evaluate the effect of parameter variation on the specific absorption rate (SAR) distribution. The performance of the new applicator type is compared with the Coaxial TEM. Operating frequency appears to be the main parameter with a positive influence on the performance. A SAR increase in tumour of 1.7 relative to the Coaxial TEM system can be obtained with a three-ring, six-antenna per ring cavity slot applicator operating at 150 MHz. PMID- 11474935 TI - The START trial-measurements in semi-anatomical breast and chest wall phantoms. AB - This paper describes dosimetry measurements performed prior to departments entering patients into the START Trial, a breast fractionation trial. Absolute and relative doses were measured in semi-anatomical breast and chest wall phantoms, as part of a quality assurance programme visit. Doses were measured using an ionization chamber and the resulting distributions were compared with those calculated by the department. The mean ratio of measured to calculated dose at the START reference point was found to be 0.981 for the breast phantom and 0.978 for the chest wall phantom. This average measured dose was significantly less than the prescribed dose (p < 0.001). Differences were found between 2D and 3D planning systems and for departments using cobalt 60 beams. A number of departments had deviations of greater than 4%, which was the tolerance applied for this trial. It is essential for dose measurements of this type to be performed for randomized clinical trials involving radiotherapy, particularly where dose fractionation regimes are being compared. PMID- 11474936 TI - Effects of crosslinker fraction in polymer gel dosimeters using FT Raman spectroscopy. AB - Radiation-induced chemical changes in polyacrylamide gels (PAGs) used in 3D radiation dosimetry have been studied using Fourier transform Raman spectroscopy. Consumption of monomer and crosslinker were characterized for polymer gel dosimeters with initial fraction of crosslinker (%C) varying from to 100%. Results indicate that the rate of monomer/crosslinker consumption is highly nonlinear over %C. This indicates that maximum polymer gel dosimeter sensitivity (i.e. slope of the monomer consumption curves) is dependent not only on initial %C but also on dose range. For low-dose regions (<5 Gy) 30%C PAGs are the most sensitive dosimeters, their maximum sensitivity being a factor of 1.8 times greater than 50%C PAGs. In mid-dose range (S-15 Gy), however, 50%C PAGs are the most sensitive dosimeters, being potentially a factor of 1.3 more sensitive than any other PAG studied. In high-dose regions (> 15 Gy) 70%C PAGs are at least 1.5 times more sensitive than any other PAG studied. A qualitative model is presented which helps to explain the variation in overall monomer consumption rate as a function of initial %C. The variation in the shapes (i.e. mathematical course) of the consumption curves for different %C PAGs is, in part, qualitatively explained by consideration of the average number of monomer to crosslinker molecules available for reaction in each PAG as it is irradiated to higher doses. Finally, we demonstrate the importance of an existing polymer network on the subsequent monomer consumption rate in a 50%C PAG. PMID- 11474937 TI - Absorbed fraction and dose conversion coefficients of alpha particles for radon dosimetry. AB - The sensitivity to different relevant parameters of the absorbed fraction of alpha particles emitted from the 222Rn chain in sensitive cells of the tracheo bronchial tree have been investigated. The structure of the airway wall given by ICRP (ICRP66) has been adopted and employed in the present calculations. The source thickness (mucous gel and sol + cilia), target layer thickness and the depth of the sensitive layers have been varied within reasonable ranges around the default values recommended by ICRP66. The results have shown that the depth of the sensitive layers is the most important parameter in calculating the absorbed fraction. In addition, dose conversion coefficients were calculated and presented along with the absorbed fractions. PMID- 11474938 TI - Phantom dosimetry for conformal stereotactic radiotherapy with a head and neck localizer frame. AB - Linear accelerator based stereotactic radiotherapy (SRT) with the Gill-Thomas Cosman (GTC) re-locatable frame has been in use for several years. The use of the frame is limited to treating lesions above the hard palate. For treating tumours in the head and neck region, the Head and Neck Localizer (HNL) frame has been designed by Radionics Inc. for use with their XPlan treatment planning software. In this study we have used a spherical acrylic phantom commercially known as the 'Lucy' phantom (Sandstrom Sandstrom Trade and Technology Inc.) to perform thermoluminiscent as well as film dosimetry for the HNL frame. A radio-opaque marker was placed in the phantom and a film test carried out to verify the accuracy in isocentre positioning. The results of the dosimetry with TLD were within 2% for points near the isocentre and 5% (or 2 mm in steep gradients) in the planning target volume (PTV). In regions of low dose, larger percentage differences in local dose were observed, but all differences were within 5% of isocentre dose. The film dosimetry provided dose distributions that matched well with those generated by the XPlan stereotactic treatment planning software. PMID- 11474939 TI - The measurement of kappach factors for application with the IPEMB very low energy dosimetry protocol. AB - In 1996. the IPEMB issued a new code of practice detailing the procedures by which the output of therapeutic kilovoltage x-ray devices are to be determined. For x-ray beam qualities in the range 0.035-1.0 mm Al half-value thickness (HVL), the equation for converting the instrument reading into absorbed dose to water contained a factor named kch. This was included to account for the change in response of the parallel plate chamber from its calibration conditions free in air to those in the user's measurement phantom. As no data were available with which to quantify this factor at very low energies, the code advised that the user take a value of unity until sufficient data became available upon which to base an addendum. In this work, kch values have been determined for four chambers: two PTW 23344 large volume soft x-ray chambers, one PTW 23342 small volume soft x-ray chamber and a Markus electron chamber. Variations in the value of kch were investigated for changes in FSD, applicator size and beam quality. The water equivalence of the Mix-D phantom used for these measurements was also verified. A comparison of the results for the two PTW 23344 chambers showed no significant differences for any experimental situation, indicating that kch is a factor of chamber design rather than variations in construction. No variation in kch was identified with changes in FSD. A small dependence on applicator size was identified for larger applicators, and this was found to be dependent upon chamber design. The measured values of kch were found to increase with energy and again differences were seen between chamber designs. Overall, the values of kch recorded during these measurements ranged from 1.01 to 1.08. These results highlight a significant underestimation of doses calculated using the very low energy code of practice. This supports the need for further work to confirm these results. and the production of an addendum to the code in its present form. PMID- 11474940 TI - A variable fluence step clustering and segmentation algorithm for step and shoot IMRT. AB - A step and shoot sequencer was developed that can be integrated into an IMRT optimization algorithm. The method uses non-uniform fluence steps and is adopted to the constraints of an MLC. It consists of a clustering, a smoothing and a segmentation routine. The performance of the algorithm is demonstrated for eight mathematical profiles of differing complexity and two optimized profiles of a clinical prostate case. The results in terms of stability, flexibility, speed and conformity fulfil the criteria for the integration into the optimization concept. The performance of the clustering routine is compared with another previously published one (Bortfeld et al 1994 Int. J. Radiat. Oncol. Biol. Ph.vs. 28 723-30) and yields slightly better results in terms of mean and maximum deviation between the optimized and the clustered protile. We discuss the specific attributes of the algorithm concerning its integration into the optimization concept. PMID- 11474941 TI - Analysing collimator structure effects in head-scatter calculations for IMRT class fields using scatter raytracing. AB - The frequent blocking of the irradiated volume in intensity modulated radiation therapy (IMRT) makes the head-scatter fraction of the incident photon fluence more significant than that in conventional therapy with open fields. On the other hand. certain collimator configurations block scatter photons directed to a given observation point while allowing primary photons to be transmitted. The 'anomalous blocking' makes the primary field a poor indicator of the scatter fluence. Since large MU-to-cGy ratios in IMRT can magnify head-scatter uncertainties, it becomes necessary to accurately model both the effective scatter source and the collimator structure that limits the scatter reaching the irradiated volume. First we obtain a dual-source model, using a Taylor series expansion to derive the effective scatter source distribution from the data measured for the Elekta SL20 linac equipped with a multi-leaf collimator (MLC). Then, using a raytracing algorithm, we calculate the transmission of scatter rays from the effective scatter source plane to points in the patient plane. The method can account for the anomalous blocking of scatter by the MLC leaves and the backup diaphragms. For a variety of collimator settings tested, the calculations agree with measurements to an accuracy of 0.002psi10 x 10, where psi10 x 10 is the total (primary + scatter) photon fluence of an open 10 x 10 cm2 field for the same MU delivered. Although the significance of collimator structure in IMRT depends strongly on fields shapes employed for the delivery, potential cumulative errors on the order of a few per cent can be avoided in fluence calculations if the proposed method is used. PMID- 11474942 TI - Monte Carlo calculation of dose rate distributions around the Walstam CDC.K-type 137Cs sources. AB - Basic dosimetric data for the Walstam CDC.K-type low dose rate 137Cs sources in water have been calculated using Monte Carlo techniques. These sources, CDC.K1 K3 and CDC.K4, are widely used in a range of applicators and moulds for the treatment of intracavitary and superficial cancers. Our purpose is to improve existing data about these sources using the Monte Carlo simulation code GEANT3. Absolute dose rate distributions in water have been calculated around these sources and are presented as conventional 2D Cartesian look-up tables. Also the AAPM Task Group 43 formalism for dose calculation has been applied. The calculated dose rate constant for the CDC.K1-K3 source is A = 1.106 +/- 0.001 cGy h(-1) U(-1), and for the CDC.K4 source, A = 1.092 +/- 0.001 cGy h(-1) U(-1). The anisotropy of the sources are accurately studied and F(r, theta) tables are given. Also phi an(r) factors are presented. The radial dose functions are given as a polynomial fit to the calculated data up to 15 cm. Best-fit values of coefficients suitable for use in Sievert integral calculations have been derived. PMID- 11474943 TI - A monte carlo study of dose rate distribution around the specially asymmetric CSM3-a 137Cs source. AB - The CSM3 137Cs type stainless-steel encapsulated source is widely used in manually afterloaded low dose rate brachytherapy. A specially asymmetric source, CSM3-a, has been designed by CIS Bio International (France) substituting the eyelet side seed with an inactive material in the CSM3 source. This modification has been done in order to allow a uniform dose level over the upper vaginal surface when this 'linear' source is inserted at the top of the dome vaginal applicators. In this study the Monte Carlo GEANT3 simulation code, incorporating the source geometry in detail, was used to investigate the dosimetric characteristics of this special CSM3-a 137Cs brachytherapy source. The absolute dose rate distribution in water around this source was calculated and is presented in the form of an along-away table. Comparison of Sievert integral type calculations with Monte Carlo results are discussed. PMID- 11474944 TI - An intercomparison of IMRT delivery techniques: a case study for breast treatment. AB - Intensity-modulated radiotherapy beams can be delivered using a multileaf collimator by one of two methods: either by superposition of a series of multiple static fields, or by moving the collimators while the beam is on to produce 'dynamically' modulated beams. The leaf trajectories in this dynamic mode are given by a series of linear steps between control points defining each collimator position at known intervals throughout an exposure. The complexity of the resulting modulation is limited in the first case by the number of fields superposed and in the second case by the number of control points defined. Results are presented for an experimental study that investigates the effect of changing both the number of fields for the multiple-static technique, and the number of control points for a dynamic 'close-in' technique. All deliveries studied are clinical intensity-modulated breast fields. The effect of using a universal wedge in conjunction with the multileaf collimator is also studied, together with a comparison of the relative efficiency, time taken and the absolute dosimetric accuracy of the various delivery options. It is shown that all delivery techniques produce equivalent dose distributions when using 15 control points, with 10 control points being sufficient to produce an adequate breast compensator distribution. Except for the case of a four-control-point dynamic delivery, the universal wedge makes no significant difference to the dose distribution. However, it makes the delivery less efficient. The close-in interpreter consistently produces deliveries that are more efficient than the more conventional sliding-window technique and faster than the multiple-static field technique. Finally the close-in technique is compared to the more 'standard' leaf-sweep technique and shown to be equivalent. PMID- 11474945 TI - A simple method to control aspects of fluence modulation in IMRT planning. AB - Many inverse-planning algorithms and commercial systems generate intensity modulated beam profiles that have considerable structure. This is the desirable outcome of the quest for high dose-space conformality. However, when these profiles are realized experimentally using the dynamic multileaf collimator (DMLC) method of delivery the monitor-unit efficiency can be quite small, with unwanted consequences. Also the interpretation of these fields leads to the generation of small field segments, again with undesirable consequences. In this note it is shown that the features of beam-space can be user-controlled to minimize these problems. There is a tradeoff between obtaining desirable features in beam-space and high conformality in dose-space. PMID- 11474946 TI - Competition no obstacle in a new cancer project created by consortium. AB - Cancer hospitals always are quick to explain that they are different, often pointing to their special commitment to their patients and ability to deal with end-of-life issues. But those differences are rarely accounted for in the benchmarking data they use. The facilities have a keen sense of competition and have, until recently, been unwilling to come together for benchmarking purposes. But all that changed in 1997 when the Joint Commission on Accreditation of Healthcare Organizations started talking about ORYX. Cancer hospital executives knew from experience that comparing data with noncancer hospitals wouldn't be useful, so a group of them came together and created a National Cancer Database initiative that would allow for subcomparisons within the ORYX system. PMID- 11474947 TI - UHC records project is attention grabber. AB - Talk about benchmarking in general and most hospitals will probably think of clinical quality indicators or financial comparisons. But for the University HealthSystem Consortium of Oak Brook, IL, a recently completed project has benchmarked medical records practices at 37 member facilities. PMID- 11474948 TI - Pain management: the fifth vital sign. AB - The Joint Commission on Accreditation of Healthcare Organizations blitzed the health care market in December to announce new pain standards. They were developed over two years in conjunction with the University of Wisconsin, Madison School of Medicine, to ensure all patients the right to appropriate assessment and management of their pain. The standards underline that organizations have a responsibility to develop processes within their settings to help support improvements in pain management. PMID- 11474949 TI - On-line FAQs (frequently asked questions) help. AB - Mark Hammerschmidt, RN, has seen plenty of orientation programs come and go during his 21 years of nursing. But when his large, New England teaching hospital lost its clinical educator for the medical intensive care unit, Hammerschmidt was asked to help out. PMID- 11474950 TI - Effects of muscle strengthening and physical conditioning training on temporal, kinematic and kinetic variables during gait in chronic stroke survivors. AB - The purpose of this study was to evaluate the impact of a combined program of muscle strengthening and physical conditioning on gait performance in subjects with chronic stroke, using a single group pre- and post-test design. Thirteen subjects were recruited for the 10-week program (3 days/week), which consisted of warm-up, aerobic exercises, lower extremity muscle strengthening and cool-down. Data from cinematographic film and a force plate obtained during multiple walking trials were used in a four-segment kinetic model to yield spatiotemporal, kinematic and kinetic variables. Gait analysis revealed that the 10 week training resulted in significant increases in gait speed associated with improvements in walking patterns as determined by increases in selected kinematic and kinetic measures. After training, subjects were able to generate higher levels of powers and demonstrated increases in positive work performed by the ankle plantar flexor and hip flexor/extensor muscles. PMID- 11474951 TI - An initial investigation of the reliability of the Rivermead Extended ADL index in patients presenting with neurological impairment. AB - The objective of this study was to establish the reliability and sensitivity of both postal and interviewer-administrated versions of the Rivermead Extended Activities of Daily Living (READL) index, which assesses six domestic activities and six community activities. Sixty patients with stable neurological impairment were recruited. In one group (n = 40), every patient was assessed face-to-face using the READL, the Barthel index (BI) and the short orientation memory and concentration test (SOMC). One week later, the READL was repeated by the same person, in the same place. In the second group (n = 20), all the patients were first sent a postal form of the READL and were then seen face-to-face for assessment as in group 1. To be included patients had to score at least 18/28 points on the SOMC. Scores were compared using scatterplots, Bland and Altman plots and correlation coefficients, and difference scores were calculated. Sensitivity was established comparing groups of patients expected to differ in their activities. Repeated assessment score, both face-to-face and by post, showed significant correlation (Pearson coefficient = 0.97 and 0.88, respectively). Most scores were within four points of each other, with no systematic bias, although patients tended to rate themselves more independent. Both methods were able to detect differences in the level of activities as predicted between more and less dependent groups (t-test: p < 0.00001 and p = 0.00087). The READL index appears to be a reliable and sensitive measure, with some evidence for validity, but further research is needed. PMID- 11474952 TI - Factors associated with successful vocational rehabilitation in a Swedish rural area. AB - The objective of this study was to identify factors associated with a positive outcome of vocational rehabilitation, and to identify groups that have been successfully rehabilitated in a Swedish rural area. In this study vocational rehabilitation is defined as medical multidisciplinary, psychological, social and occupational activities aiming to re-establish, among sick or injured people with previous work history, their working capacity and prerequisites for returning to the labour market. The study was based on 732 people on registered long-term sick leave who, in a rural area in northern Sweden during 1992-94, became objects for vocational rehabilitation. Bivariate and stepwise logistic regression analysis was used to identify factors associated with the outcome. By successful vocational rehabilitation is meant reporting well (no economical benefit) at all three time-points 6, 12 and 24 months after termination of rehabilitation, or lowered benefit levels. The results indicate that younger, male, employed persons, with an early start on rehabilitation, in a programme entailing education, and partly sick-listed before the start of this programme, had the greatest chance of successful rehabilitation. In contrast, older, female, unemployed people, with a delayed start on rehabilitation, without education, and fully sick-listed before the start, greatly risked being unsuccessful with vocational rehabilitation. The results indicate how to improve the rehabilitation process: several process-related factors shown to be connected with successful vocational rehabilitation include time before the start of rehabilitation, partial instead of full sickness benefit, and education programmes. PMID- 11474953 TI - Chest physiotherapy after coronary artery bypass graft surgery--a comparison of three different deep breathing techniques. AB - The effectiveness of three deep breathing techniques was evaluated in 98 male patients after coronary artery bypass graft surgery in a randomized trial. The techniques examined were deep breathing with a blow bottle-device, an inspiratory resistance-positive expiratory pressure mask (IR-PEP) and performed with no mechanical device. Pulmonary function and roentgenological changes were evaluated. Four days post-operatively there were significantly decreased vital capacity, inspiratory capacity, forced expiratory volume in 1 second, functional residual capacity, total lung capacity and single-breath carbon monoxide diffusing capacity in all three groups (p < 0.0001). No major differences between the treatment groups were found, but the impairment in pulmonary function tended to be less marked using the blow bottle technique. The Blow bottle group had significantly less reduction in total lung capacity (p = 0.01) compared to the Deep breathing group, while the IR-PEP group did not significantly differ from the other two groups. PMID- 11474954 TI - Torque variations during repeated passive isokinetic knee movements in persons with multiple sclerosis. AB - The purposes of this study were to investigate the effect of movement repetitions on resistive torque during passive isokinetic dynamometry of the knee and to determine the role of electromyographic activity in the stretched muscles on the torque measurements. Ten persons with multiple sclerosis and hypertonia of the knee muscles were compared with 10 healthy age- and gender-matched control subjects. During series of 10 flexion and extension movements of the knee at 60, 180 and 300 degrees/s, torque and electromyographic activity in the stretched muscles were registered. The persons with hypertonia presented a significantly larger torque reduction (p < 0.05) than the control subjects in all test conditions except for repeated knee flexion at 300 degrees/s. Electromyographic activity in the stretched muscles was not identified as the only explanatory mechanism for the reduction in hypertonia during the movement repetitions, suggesting that other factors were also involved. PMID- 11474955 TI - Ankle dorsiflexor muscle performance in healthy young men and women: reliability of eccentric peak torque and work measurements. AB - The aims of this study were: (i) to assess the test-retest intrarater reliability of eccentric ankle dorsiflexor muscle performance in young healthy men and women using the Biodex dynamometer; and (ii) to examine different statistical indices for the interpretation of reliability. Thirty men and women (age 22.5 +/- 2.5 years, mean +/- S.D.) performed three maximal eccentric contractions at 30 degrees/second and 90 degrees/second, with 7-10 days between test sessions. Reliability was evaluated with three intraclass correlation coefficients (ICC1,1, ICC2,1 and ICC3,1), and was excellent for peak torque (ICC 0.90-0.96) and good to excellent for work (ICC 0.69-0.83), with no discernible differences among the three ICCs. Method errors, assessed by the standard error of the measurement (S.E.M.) and S.E.M.%, were low. The Bland & Altman graphs and analyses indicated no significant systematic bias in the data. In conclusion, measurements of eccentric ankle dorsiflexor muscle performance in young healthy individuals using the Biodex are highly reliable. PMID- 11474956 TI - Reducing sedentary behavior: role in modifying physical activity. AB - Decreased physical activity is associated with the increased incidence of obesity. Behavioral economic research demonstrates that reducing sedentary behaviors in children increases physical activity. Understanding how people choose physical or sedentary activities can aid in developing public health initiatives that increase access to physical activity, while reducing access to sedentary behaviors. PMID- 11474957 TI - Sex differences in human skeletal muscle fatigue. AB - Research on muscle fatigue suggests that greater fatigue resistance may be evident in females compared with males. The possible mechanisms for this sex difference include factors related to muscle mass, substrate utilization, muscle morphology, and neuromuscular activation. PMID- 11474958 TI - Hyponatremia associated with exercise: risk factors and pathogenesis. AB - Exercise-related hyponatremia is an infrequent but potentially life-threatening accompaniment of prolonged exercise. This condition results from sodium losses in sweat, excessive water intake, or both. We review the risk factors for development of this condition and discuss evidence that there is a population at increased risk of hyponatremia during prolonged exercise. PMID- 11474959 TI - Does aging adversely affect muscle mitochondrial function? AB - Muscle oxidative function appears to decline with aging, and evidence suggests that this is related to reduced synthesis of mitochondrial and other muscle proteins. Causes for these events may include mtDNA damage or reduced mtDNA copy numbers, reduced oxidative enzyme activities and ATP production, and increased proton leak. PMID- 11474960 TI - Visual expertise in the perception of action. AB - Within the exercise and the sports sciences, various "expert observers" rely on visual assessments of movement. Strengths and limitations of unaided observation are outlined based on the biological motion perception literature and related work. The concepts of coordination, control, and action are used to propose an "action prototype" for pattern recognition. PMID- 11474961 TI - Regulation of oxygen consumption at the onset of exercise. AB - Increased aerobic production of ATP at the onset of exercise could be limited by availability of metabolic substrates independent of O2, or interaction between O2 and metabolic substrates. We point out the importance of feedback control to match O2 supply to demand and discuss metabolic control at the onset of exercise. PMID- 11474962 TI - Regulation of oxygen consumption at exercise onset: is it really controversial? AB - The conflicting hypotheses on the limiting factors for skeletal muscle VO2 on kinetics might be reconciled in a unifying scenario. Under "normal" conditions, during transitions to moderate intensity exercise, the limiting factor appears to be an inertia of oxidative metabolism. During transitions to exercise of higher metabolic intensity, O2 delivery could play a relatively minor but significant role as a limiting factor. PMID- 11474963 TI - Why do people jump the way they do? AB - When humans perform maximum height squat jumps, their segmental rotations contribute in a proximodistal sequence to the vertical acceleration of the center of gravity. The same kinematic pattern occurs in a forward dynamic model of the musculoskeletal system when muscle stimulation is optimized to maximize jump height. This paper examines why this kinematic pattern maximizes jump height in humans, given the design of the human musculoskeletal system. PMID- 11474964 TI - A reliable gait phase detection system. AB - A new highly reliable gait phase detection system, which can be used in gait analysis applications and to control the gait cycle of a neuroprosthesis for walking, is described. The system was designed to detect in real-time the following gait phases: stance, heel-off, swing, and heel-strike. The gait phase detection system employed a gyroscope to measure the angular velocity of the foot and three force sensitive resistors to assess the forces exerted by the foot on the shoe sole during walking. A rule-based detection algorithm, which was running on a portable microprocessor board, processed the sensor signals. In the presented experimental study ten able body subjects and six subjects with impaired gait tested the device in both indoor and outdoor environments (0-25 degrees C). The subjects were asked to walk on flat and irregular surfaces, to step over small obstacles, to walk on inclined surfaces, and to ascend and descend stairs. Despite the significant variation in the individual walking styles the system achieved an overall detection reliability above 99% for both subject groups for the tasks involving walking on flat, irregular, and inclined surfaces. In the case of stair climbing and descending tasks the success rate of the system was above 99% for the able body subjects and above 96 % for the subjects with impaired gait. The experiments also showed that the gait phase detection system, unlike other similar devices, was insensitive to perturbations caused by nonwalking activities such as weight shifting between legs during standing, feet sliding, sitting down, and standing up. PMID- 11474965 TI - Increased risk for falling associated with obesity: mathematical modeling of postural control. AB - Recent epidemiological studies report that obesity is positively related to fracture incidence. In the present experiment, a model of postural control was used to examine the impact of an abnormal distribution of body fat in the abdominal area upon postural stability. Obese and lightweight humanoids were destabilized by imposing a small initial angular speed from a neutral standing position. To avoid a loss of stability yielding a stepping reaction or a fall, an ankle torque is necessary to counteract the perturbation. Three torque parameters -ankle torque onset, time to peak torque, and muscular ankle torque--were entered in a program to simulate the intrinsic variability of the human postural control system. A loss of stability was detected when the center of pressure exceeded stability margins. The most striking observation is the nonlinear increase of torque needed to stabilize the humanoid when the motor response was characterized by delayed temporal parameters. The effect was more pronounced when an anterior position of the center of mass was included in the simulations. This suggests that, when submitted to daily postural stresses and perturbations, obese persons (particularly those with an abnormal distribution of body fat in the abdominal area) may be at higher risk of falling than lightweight individuals. PMID- 11474966 TI - Modeling the postural disturbances caused by upper extremity movements. AB - This paper describes the design, validation, and application of a dynamic, three dimensional (3-D) model of the upper extremity for the purpose of estimating postural disturbances generated by movements of the arms. The model consists of two links representing the upper and lower arms, with the shoulder and elbow modeled as gimbal joints to allow three rotational degrees of freedom. With individualized segment inertial parameters based on anthropometric measurements, the model performs inverse dynamic analysis of recorded arm movements to calculate reaction forces and moments acting on the body at the shoulder in three dimensions. The method was validated by comparing the output of the model to estimates obtained from ground reaction loads during stereotypical and free form unilateral movements at various velocities and with different loads carried by human subjects while seated on biomechanical force platforms. The correlation between predicted and measured reaction forces and moments was very good under all conditions and across all subjects, with average rms errors less than 8% of measured peak-to-peak values. The model was then applied to bimanual activities representative of functional movements that would typically be performed while standing at a counter. The resulting estimates were consistent and adequate for the purpose of evaluating postural disturbances caused by upper extremity movements. PMID- 11474967 TI - Characteristics of somatosensory feedback in postural control during standing. AB - In the present study, the function of the somatosensory feedback system in postural control was investigated. For the sake of simplicity, the present study considered only balancing in the anteroposterior direction using the ankle strategy, in which the ankle moment is mainly used to maintain balance. To suppress the vestibular and visual feedback paths, a subject stood on a force measuring platform with a fixed back support. Because the subject's body was immovable under these conditions, the subject controlled a computer model that simulated the subject's load at the ankles. Information about the sway angle of the model was fed through the somatosensory feedback path. Frequency response functions of the ankle moment in response to the sway angle were calculated. The experimental results suggest that the human somatosensory feedback system has derivative characteristics and, consequently, can maintain an upright posture by itself. The results were compared with those of previous studies on vestibular and visual feedback systems. The comparison reveals that subject-to-subject variance in the somatosensory system is significantly smaller than that in the other systems. This may indicate that the somatosensory feedback is the most automatic of the systems and plays a dominant role when a subject maintains an upright posture using the ankle strategy. PMID- 11474968 TI - Somatotopy of the motor cortex after long-term spinal cord injury or amputation. AB - Certain brain-computer interface (BCI) methods use intrinsic signals from the motor cortex to control neuroprosthetic devices. The organization of the motor pathways in those populations likely to use neuroprosthetic devices, therefore, needs to be determined; there is evidence that following disease or injury the representation of the body in the motor cortex may change. In this study, functional MRI measures of somatotopy following spinal cord injury (SCI) showed evidence of changes in limb representations in the motor cortex. Subjects with chronic SCI had unusual cortical patterns of activity when attempting to move limbs below their injury; amputees showed a more normal somatotopy. The functional reorganization may affect optimal implanted electrode placements for invasive BCI methods for these different populations. PMID- 11474969 TI - A pilot study of myoelectrically controlled FES of upper extremity. AB - Functional electrical stimulation (FES) of upper limbs can be used for the recovery of some hand functions on patients with CNS lesions. This study deals with the control of FES by means of myoelectrical activity detected from voluntarily activated paretic muscles. The specific aim of this paper is to evaluate the accuracy of myoelectrical control in terms of produced force and movement. For this purpose, a specific device called myoelectrical controlled functional electrical stimulator (MeCFES) has been developed and applied to six tetraplegic patients with a spinal cord lesion and one stroke hemiplegic patient. Residual myoelectric signals from the paretic wrist extensor (m. extensor carpi radialis, ECR) have been used to control stimulation of either the wrist extension (i.e., the same muscle) or thumb flexion. A tracking test based on a visual feedback of the produced force or movement compared to a reference target trajectory was used to quantify control accuracy. A comparison was made between the tracking performances of each subject with and without the MeCFES and the learning process for two of the subjects were observed during consecutive sessions. Results showed that the wrist extension was improved in three out of five C5 SCI patients and the thumb flexion was largely increased in one incomplete C3 SCI patient. The hemiplegic patient showed limited thumb control with the MeCFES but indicated the possibility of a carry over effect. It was found that a low residual natural force resulted in a less accurate movement but also with a large increase (up to ten times) of the muscle output. On the contrary, persons with a medium residual force obtained a smaller amplification of muscle force with a higher tracking accuracy. PMID- 11474970 TI - Test bed with force-measuring crank for static and dynamic investigations on cycling by means of functional electrical stimulation. AB - Cycling by means of functional electrical stimulation (FES) is an attractive training method for individuals with paraplegia. The physiological benefits of FES are combined with the psychological incentive of independent locomotion. In addition, cycling has the advantage in that the generated muscle forces are converted into drive power with relatively high efficiency compared to other means of locomotion, e.g., walking. For the design of an appropriate cycling device and the development of optimal stimulation patterns, it has to be investigated how the geometry for FES cycling, influenced by individual parameters of the FES-generated drive torques and the magnitude of variations among subjects with paraplegia, can be optimized. This study shows the design of a freely adjustable test bed with additional motor drive which allows static and dynamic measurements of force components and drive torque at the crank. Furthermore, the influence of geometry and various individual parameters on FES pedaling can be tested for each subject individually. A pedal path realized by a three-bar linkage that was optimized according to preliminary simulations further increases leg cycling efficiency. Safety precautions avoid injuries in case of excessive forces, e.g., spasms. Test results illustrate the application of the test bed and measurement routines. A test series with four paraplegic test persons showed that the presented static and dynamic measurement routines allow to provide optimal stimulation patterns for individual paraplegic subjects. While pedaling with these optimal stimulation patterns only negligible negative active drive torques, due to active muscle forces, were applied to the crank and sufficient drive power was generated to power a cycle independently. PMID- 11474971 TI - Localization and control of a rehabilitation mobile robot by close human-machine cooperation. AB - In the field of rehabilitation robotics, a mobile personal robot represents an attractive solution, especially in economic terms in comparison with a desktop workstation. A manipulator arm mounted on a mobile robot can facilitate the restoration of the disabled user's manipulative function. In order both to encourage the person to participate in the task at hand and to be cost effective, close human-machine cooperation is essential. The person controls the robot via a remote station and develops strategies to successfully carry out a mission. The main problems encountered by the person during the execution of a mission are electing to change modes, and the mode transition itself. We have examined two aspects of this cooperation: 1) information exchange between human and machine for decision-making and 2) giving to operators complementary and redundant modes to command the system. An experiment has been conducted to study these two aspects. This paper focuses on the control of robot movements in an indoor environment and especially on localization parameters, human-like robot behavior, and the value of proposing complementary control modes to the operator. PMID- 11474972 TI - A fuzzy rule-based approach to recognizing 3-D arm movements. AB - One particular application of gesture-based systems is to implement a speaking aid for the deaf. For this to happen, it requires a module that can recognize three-dimensional (3-D) arm movements since arm movement is one of the four main attributes characterizing a sign word. In this paper we propose a fuzzy rule based recognition method. The effectiveness of this system is evaluated by recognizing 3-D arm movements involved in Taiwanese Sign Language (TSL). PMID- 11474973 TI - Surgical rehabilitation of the planovalgus foot in cerebral palsy. AB - The objectives of this study were to quantitatively determine the effects of subtalar arthrodesis on the planovalgus foot using three-dimensional (3-D) gait analysis and plantar pressure measurements. Twelve children and adolescents with planovalgus foot deformity secondary to spastic cerebral palsy participated in this outcome study. The pediatric population were evaluated preoperatively and following subtalar fusion. Seventeen feet were operated for the correction of the planovalgus foot deformity. A Holter-type microprocessor-based portable in-shoe data acquisition system was used in this study to collect the multistep dynamic plantar pressure history, while a five-camera Vicon-based gait analysis system was used to track the lower extremity joint kinematics. The results obtained from the plantar pressure measurement showed significant increases in mean peak vertical plantar pressures postoperatively at the lateral midfoot and lateral metatarsal heads. Mean contact durations and mean pressure-time integrals were also significantly increased at these plantar locations following foot surgery. This redistribution in pressure metrics suggests the formation of new lateral plantar weight bearing areas. The 3-D gait analysis system, using standardized lower extremity measurements, was unable to reveal any significant changes in joint kinematics, particularly at the foot and ankle where the surgery was performed. This suggests the need for a more refined system to track the complex motion of the pediatric foot and ankle during gait. PMID- 11474974 TI - Analysis of sliding and pressure distribution during a repositioning of persons in a simulator chair. AB - This study was undertaken to investigate the effect of system tilt and back recline angles on sliding and pressure distribution of seated subjects. Ten able bodied subjects adopted successively 12 postures on a multiadjustable simulator chair. The system tilt angle was varied from 0 degrees to 45 degrees posterior tilt, while the seat to back angle varied from 90 degrees to 120 degrees. A maximum of 40.2% of weight shift was found when combining a system tilt angle of 45 degrees to a seat to back angle of 120 degrees. Maximum value of 74 mm of sliding was observed for the acromion marker during repositioning. Significant weight shift at the level of the seat is obtained only when the system tilt angle exceeds 15 degrees in a posterior direction. We can put forward here that a small tilt < or =15 degrees can be used to adjust back pressure distribution, whereas large posterior tilts are used for an effective weight shift at the seat level. The peak pressure gradient remains in general in the interval of +/-30% from the neutral posture for the able-bodied subjects and is fairly constant at 15 degrees of tilt. A significant amount of displacement along the back and seat reference plane were found for the shoulder and hip markers, but this displacement does not necessarily correspond to a pure translation motion of the pelvic segment. PMID- 11474975 TI - Fuzzy-control of a hand orthosis for restoring tip pinch, lateral pinch, and cylindrical prehensions to patients with elbow flexion intact. AB - This study examines, through simulation, the use of fuzzy logic as a feasible control scheme for a hand orthosis that can restore fingertip pinch, lateral pinch, and cylindrical grasps to individuals suffering from C5-C7 spinal cord injuries. A simplified hand orthosis model, consisting of four fingers and a thumb, was derived for the purpose of planning appropriate grasp trajectories and for validating the fuzzy logic control architecture. For comparison a proportional-plus-integral-plus-derivative (PID) controller was also designed. Fuzzy logic is advantageous for this system since it eliminates solving coupled nonlinear equations of motion. For the various grips, the fuzzy controller produced better performance than the PID controller. PMID- 11474976 TI - Accuracy and reliability testing of a portable soft tissue indentor. AB - We have designed, built, and tested a portable indentor device that allows us to determine force/displacement (F/D) measurements on soft tissue in a clinical or research setting. The indentor system consists of a load cell mounted on a three dimensional measurement device (Metrecom). The output of the load cell and the Metrecom are recorded and analyzed by software running on a notebook computer. The displacement calibration of the Metrecom gave an average error = 0.005 mm [standard deviation (SD) = 0.062)]. The force calibration of the load cell resulted in an average error = 0.022 N (SD = 0.049) and a linearity of 1.0062 (R2 = 0.9998). The indentor device was tested on six different human soft tissues by two different investigators. The interreliabilities and intrareliabilities were 0.99 [interclass correlation (ICC)] indicating that the results were repeatable by more than one investigator. F/D measurements from indentor testing on two materials were comparable to values measured using an Instron device (5.34 versus. 5.52 N/mm, and 0.98 versus 1.04 N/mm). The device was used to measure the soft tissue characteristics on the plantar surface of the foot of one subject. These data were used to calculate the effective Young's modulus for the tissue using equations derived by Zheng et al. [1] and indicated a wide range of values dependent upon the portion of the F/D curve used. All results indicate data from this portable indentor device are reliable, accurate, and sensitive enough to identify mechanical properties of human tissues. PMID- 11474977 TI - Access is better for racial/ethnic elderly in Medicare HMOs--but disparities persist. PMID- 11474978 TI - Plasmodium kentropyxi n.sp. (Apicomplexa: Haemosporina: Plasmodiidae) and a Plasmodium tropiduri-like parasite in the lizard Kentropyx calcarata (Lacertilia: Teiidae) in north Brazil. AB - Plasmodium kentropyxi n.sp. is described in the teiid lizard Kentropyx calcarata from north Brazil. Young asexual stages and gametocytes are at first polar in the erythrocyte but with elongation, move to a lateral position. Largest meronts seen contained from 30-40 nuclei and conspicuous greenish-black pigment granules located in a distinct vacuole. With growth the gametocytes eventually assume a smooth, curved cylindrical shape, with evenly rounded ends. Pigment is scattered or concentrated around a conspicuous vacuole which is slowly developed as the gametocytes mature. Mature male parasites measured 11.8 x 4.0 microns (9.6 x 4.2 13.2 x 3.6 microns), shape-index 2.9 (2.2 - 5.0), and females 13.5 x 4.5 microns (12.0 x 4.5 - 15.0 x 4.8 microns), shape-index 3.0 (2.2 - 3.8). Some larger meronts may slightly enlarge the erythrocyte, but most asexual stages and the mature gametocytes rarely do so. A second, P. tropiduri-like parasite encountered in K. calcarata possessed small rounded or fan-shaped meronts producing from 4-14 merozoites, and spherical to subspherical gametocytes of approximately 6.0 x 5.0 microns. The parasite was consistently polar in its position in the erythrocyte. PMID- 11474979 TI - Life history and pathogenesis of Gallegostrongylus australis (Nematoda: Angiostrongylidae) in muridae. AB - Gallegostrongylus australis Spratt, Haycock & Walter, 2001 (Nematoda: Angiostrongylidae) developed in Deroceras panormitanum, Lehmannia nyctelia, L. flava and Milax gigates (Gastropoda). The first moult occurred at 18-19 days after infection (DAI) and the second moult at 28 DAI. Larvae were infective to experimental murid definitive hosts at 35 DAI. In experimentally infected Rattus fuscipes larvae moulted L3-4 at 3 DAI and L4-5 at 6-7 DAI. Patency in R.fuscipes, R. lutreolus, R. norvegicus and R. rattus occurred 27-64 DAI and duration varied from 7-392 days. Histopathological changes in the lungs of R. lutreolus and development of debilitating clinical signs, in contrast to R. fuscipes, suggests that the former host-parasite relationship may be the more recent one but other traits suggest the opposite. Patent infections were established in some wild R. rattus and some laboratory R. norvegicus but not in wild M. domesticus, laboratory M. musculus, rabbit, Oryctolagus cuniculus, and marsupial bandicoot, Isoodon macrourus. PMID- 11474980 TI - Monogeneans from Pangasiidae (Siluriformes) in Southeast Asia: I. Five new species of Thaparocleidus Jain, 1952 (Ancylodiscoidinae) from Pangasius pangasius, P. kinabatanganensis, P. rheophilus and P. nieuwenhuisii. AB - The examination of gill parasites from Pangasius pangasius (Hamilton, 1822); P. kinabatanganensis Roberts & Vidthayanon, 1991; P. rheophilus Pouyaud & Teugels, 2000 and P. nieuwenhuisii (Popta, 1904) (Siluriformes, Pangasiidae) in Southeast Asia revealed the presence of six species of Thaparocleidus Jain, 1952 (Monogenea, Ancylodiscoidinae). One has been previously described from P. pangasius: T. pangasi (Tripathi, 1957). The other five are new species: T. chandpuri n. sp. from P. pangasius; T. bahari n. sp. and T. sabanensis n. sp. from P. kinabatanganensis; T. redebensis n. sp. from P. rheophilus and T. mahakamensis n. sp. from P. nieuwenhuisii. PMID- 11474981 TI - Chromosome studies of Ichthyocotylurus platycephalus (Creplin, 1825) Odening 1969 with description of triploid variant and comparative karyology of the genus Ichthyocotylurus. AB - This paper reports the first karyological study of trematode Ichthyocotylurus platycephalus (Creplin, 1825) Odening 1969. Chromosome number and morphology were studied in somatic cells of parthenites from intermediate host--mollusc Valvata piscinalis using an air drying technique and Giemsa staining. The karyotype consisted of 20 chromosomes comprising five pairs of subtelo-acrocentric (1st to 5th), two pairs of submeta-subtelocentric (6th-7th), one submetacentric (9th) and two pairs of metacentric chromosomes (8th, 10th). A supernumerary chromosome variation (presence/absence of B-chromosome) and occurrence of triploidy in one population of I. platycephalus are reported. In addition, the karyological analysis of Lithuanian population of I. variegatus was carried out. In the light of presented and previously obtained data comparative karyology of genus Ichthyocotylurus is discussed. PMID- 11474982 TI - [Two new species of Stivalius (Siphonaptera--Pygiopsyllidae--Pygiopsyllinae) from Indonesia]. AB - The fleas, S. meridionalis n. sp. from Timor and S. franciscae n. sp. from Sulawesi, Indonesia are described and illustrated. S. meridionalis was collected in Central Timor (Timor Tengah) at elevations between 930 m and 1,520 m, mainly from the murine rodent Rattus tanezumi, although one specimen was recovered from a house shrew, Suncus murinus. S. franciscae was collected in Central Sulawesi (Sulawesi Tengah) at known elevations between 950 m and 2,255 m from six species of native endemic murine rodents, especially Rattus hoffmanni, and from two species of commensal murine rodents. One specimen of S. franciscae was also collected from the native endemic murine Paruromys dominator in North Sulawesi (Sulawesi Utara) at 1,780 m. These two new flea species are compared with the previously described taxa included in the genus Stivalius sensu stricto. PMID- 11474983 TI - Scanning electron microscopy of larval instars and imago of Oestrus caucasicus (Grunin, 1948) (Diptera: Oestridae). AB - Oestrus caucasicus (Grunin, 1948) is a larval parasite of the nasal cavities of Capra caucasica, Capra ibex and Capra pyrenaica. This study is the first description of the parasite using scanning electron microscopy. The first larval instar shows minor differences with Oestrus ovis. The second larval instar shows important synapomorphic features common to Oestrus ovis but, also, distinctive features as the spines-crown or the currycomb-shaped spines. The third larval instar shows many differences with Oestrus ovis, mostly in the ventral and dorsal spines. The imagos of the two species have closely related morphologies. This study is a contribution to a revision of phylogeny of Oestridae family. PMID- 11474984 TI - [New observation in France of a nuisance due to Simulium (Boophthora) erythrocephalum (De Geer, 1776) (Diptera, Simuliidae)]. AB - Some nuisance due to Simuliidae appeared near the French border with Spain (Pyrenees-Orientales) during the summer 1998. Larvae and pupae were collected in the lower parts of the rivers Tet, Tech and Agly. The identification demonstrated the occurrence of Simulium erythrocephalum. If the nuisance would become durable, a control campaign should be initiated. PMID- 11474985 TI - Isolation of a cDNA encoding an IgG immunoreactive antigen (zinc finger protein) of Strongyloides stercoralis. AB - A full length cDNA encoding an IgG immunoreactive antigen of Strongyloides stercoralis is described. A clone containing 1,328 bp insert was selected following screening of S. stercoralis cDNA library with an IgG fraction obtained from a pool of 78 S. stercoralis positive human sera samples. The nucleotide sequence of the 1,328 bp insert was found to be 70.5% A/T, reflecting a characteristic A/T codon bias of S. stercoralis. The nucleotide sequence of this insert identified a cDNA coding for a zinc finger protein. The conceptually translated amino acid sequence of the open reading frame for the IgG immunoreactive antigen of S. stercoralis encodes a 211 amino acid residue protein with an apparent molecular weight of 22.8 kDa and a predicted isoelectric point of 8.71. The diagnostic potential of this IgG immunoreactive antigen of S. stercoralis is also discussed. PMID- 11474986 TI - Serological differentiation of microsporidia with special reference to Trachipleistophora hominis. AB - Myositis is a common clinical syndrome in advanced stages of AIDS. Trachipleistophora hominis (phylum Microspora) has been detected in several cases of painful, immobilising myositis in AIDS patients. Enzyme linked immunosorbent assays (ELISAs) and Western blotting of protein profiles separated by SDS PAGE were used to determine whether this species could be detected and differentiated by serology. Sixteen microsporidia, including several species known to infect man and species infecting fish, crustaceans and a mosquito, were used as antigen. Each species had a unique profile of SDS PAGE-separated proteins. In Western blots, mouse antiserum, raised to T. hominis and selected for its high ELISA specificity, bound to antigens ranging from less than 25 kDa to greater than 250 kDa with major bands at 39-44 kDa and 98-150 kDa on T. hominis protein profiles. The serum also recognised some high molecular weight antigens in the profiles of Vavraia culicis, Heterosporis anguillarum, and three species of Pleistophora but none in the remaining genera examined. It was concluded that ELISA and Western blotting could be used to detect and differentiate T. hominis in muscle biopsy tissue from patients with myositis. However, sera from T. hominis-infected patients in the terminal stages of AIDS would not be useful for detection of infections because of a sharp decline in antibody level. PMID- 11474987 TI - Virulence for BALB/c mice and antigenic diversity of eight Toxoplasma gondii strains isolated from animals and humans in Brazil. AB - With the purpose of establishing alternative parameters to determine the virulence of Toxoplasma gondii strains, the antigenic diversity of eight strains of the parasite isolated in Brazil was evaluated. BALB/c mice were inoculated i.p. with 10(0), 10(1), 10(2) and 10(3) tachyzoites from each strain. The mortality and time to death of the animals showed that T. gondii strains may be divided in three groups: three strains resulted in 100% of mortality, 5-10 days post inoculation (DPI); three strains resulted in 100% of mortality, 7-19 DPI and brain cysts were observed in the mice which were inoculated; two strains resulted in 0% of mortality, 30 DPI. The analysis of the antigenic profile of different T. gondii strains through Western blotting, using rabbit antiserum to T. gondii, revealed that most antigens are similar to all strains. The mAb 4C3H4 recognized antigens only in the RH, N, AS28 and ME49 strains. PMID- 11474988 TI - [Comparison of effects of endotracheal suction and surfactant administration on hemodynamics and oxygenation of premature infants--a near infrared spectroscopy study]. AB - BACKGROUND: Aim of this study was to assess whether the decrease in heart rate and oxygen saturation occurring acutely after bolus administration of surfactant is different from the effect of endotracheal suctioning and whether there are effects on cerebral haemodynamics and oxygenation. METHODS: Twelve premature infants, birth weight 720-1560 g (Median 935 g), gestational age 25-30 weeks (Median 28 weeks), were investigated during surfactant administration (Curosurf) as a single bolus and a preceding endotracheal suctioning procedure. Using near infrared spectroscopy changes in cerebral blood volume and cerebral oxygenation index were assessed. Heart rate, arterial oxygen saturation, PCO2, and arterial blood pressure were registered continuously. Data were compared at 30, 60 and 120 seconds after the beginning of suctioning and surfactant administration. RESULTS: Endotracheal suctioning and surfactant administration induced a similar decrease in heart rate (-29/min, SD 25, p < 0.01 and -30/min, SD 23, p < 0.001) during the first 30 seconds and in oxygen saturation at the 30 seconds datapoint (-3%, SD 2, p < 0.01 and -3%, SD 6, p < 0.05). The decrease in oxygen saturation persisted at 60 seconds after the start of suctioning (-4%, SD 5, p < 0.05). In contrast, oxygen saturation was no longer different from baseline levels 60 seconds after surfactant administration (+1%, SD 6, p = 0.5). After suctioning, cerebral oxygenation index had decreased significantly from baseline levels at 30, 60, and 120 seconds. In contrast, after surfactant administration, no significant difference from baseline was noted at 30 and 60 seconds and a significant increase occurred at 120 seconds. Mean arterial blood pressure, PCO2, and cerebral blood volume did not change significantly during endotracheal suctioning and during surfactant administration. CONCLUSIONS: Acute decreases in heart rate and oxygen saturation during bolus administration of surfactant are equal to effects occurring during endotracheal suctioning. However, the decrease in oxygen saturation is shorter and more importantly, no decrease in cerebral oxygenation occurs. PMID- 11474989 TI - [Aortic dissection in pregnancy in Marfan syndrome--case report and treatment concept]. AB - PATIENTS AND METHODS: Herein we report the case of a 37 year old pregnant women with Marfan syndrome with dilatation of the aorta ascendens of 42 mm preconceptionAl. The dilatation remained stable until 34 weeks of gestation when a aortic dissection (De Bakey type I) occurRed rapidly. The patient underwent a cesarean section and hysterectomy, followed by surgery of the aorta asc. by means of implantation of a valved graft immediately. CONCLUSIONS: The criteria of a dilatation of the aorta ascendens < 40 mm does not identify reliably those patients who are at risk of dissection during pregnancy. A interdisciplinary approach together with obstetricians, cardiologists and cardiac surgeons is mandatory in all cases of Marfan syndrome and pregnancy. PMID- 11474990 TI - [Massive placental hemorrhage in partial molar pregnancy due to hypotriploidy (68,XX)--prenatal diagnosis and clinical course]. AB - BACKGROUND: Triploidy describes the presence of threefold haploid chromosome set and is a frequent cause of early abortion. Only few pregnancies reach the second trimester. CASE REPORT: Prenatal diagnosis of a fetus in the 28th week of gestation with a hypotriploidy and with characteristic ultrasonographic features is presented. An enlarged placenta with molar changes, an oligohydramnion, severe growth retardation and minor cardiac anomalies were observed by ultrasound. Before prenatal karyotyping could be performed, immediate cesarian section was necessary due to massive intraplacentar hemorrhage resulting in a decrease of the hemoglobin level. The diagnosis of triploidy was confirmed postnatally by cytogenetic analysis of lymphocytes, the child died after 3 days. DISCUSSION: Ultrasonographic and clinical features for the diagnosis of triploidy are presented. Etiology of the rare karyotype 68,XX is discussed. PMID- 11474991 TI - [Fetal obstructive uropathy--diagnosis and possible treatments]. AB - BACKGROUND: Abnormalities of the urinary tract are found in up to 5% of newborns. They account for 25% of all prenatally diagnosed congenital defects and contribute 4% to perinatal mortality. When considering the prognosis of these anomalies, association with other pathological conditions has to be taken into account. MATERIALS AND METHODS: We discuss the state of the art in detection and management of fetal urinary tract abnormalities as are standard at this tertiary center of fetal medicine. RESULTS AND DISCUSSION: Posterior urethral valve, megaureter or ureteropelvic junction obstruction represent one end of a wide spectrum of obstructive uropathies, ranging from pathophysiologically minor deviations to severe impairment of renal function. Organ development, physiology of urine production and excretion as well as their potential disorders are presented. CONCLUSION: Sonographic and invasive diagnostic options in the evaluation of the urogenital tract are discussed. Careful assessment of each individual case and prognosis determine pre- and postnatal treatment. PMID- 11474992 TI - [Comment on Beller FK. Incidence and causes of upper arm plexus paralysis in newborn infants. An overview and basic principles of legal considerations]. PMID- 11474993 TI - New avenues in surfactant research. PMID- 11474994 TI - [Oral sugar solutions in pain therapy of neonates and premature infants]. AB - OBJECTIVES: We review the efficacy of oral sugar solutions for treating procedural pain in neonates and address the following questions: Do newborns need analgesic therapy for procedural pain during blood sampling? How do sugars influence pain-reactions of neonates? What is the efficacy of sugar solutions in clinical practice? METHODS: We searched for relevant articles in the PubMed database from 1990 to September 2000. RESULTS: Treatment of procedural pain in newborns is desirable because they are more sensitive to pain than adults, they show marked pain reactions during blood sampling and repeated acute pain in the newborn period results in longterm behavioural changes. Oral sugar solutions have been studied for treatment of procedural pain in neonates. Their initial effect is the result of orotactile stimulation by the intraoral fluid. The orogustatory stimulation by the sweet taste prolongs the effect for up to 10 minutes through endorphin release. In randomized-controlled trials oral sugar solutions (2 ml of 25% sucrose or 30% glucose) reduced pain reactions and crying and attenuated the heart rate increase after capillary and venous blood sampling in term and preterm neonates. They are more effective than traditional calming strategies, like cuddling by parents, use of a pacifier, or breast feeding. Yet, sugar solutions provide no adequate analgesia for more severe pain, e.g. during circumcision. CONCLUSIONS: Sugar solutions effectively relieve procedural pain during blood sampling in neonates. Additional studies are needed to determine the minimal effective dose and the efficacy and side effects of repeated sugar doses in the same patient. PMID- 11474995 TI - [Three-dimensional fetal echocardiography--current status and future prospects]. AB - The purpose of echocardiographic imaging in the fetus is to provide clear representations of the underlying cardiac anatomy. Studies on pre- and postnatal 3D echocardiography have shown this technique can provide an alternative form of image display for comprehensive assessment of congenital heart disease. To date, several different methods are used for 3D echocardiography in the fetus. The technique used at our institution is analogous to the 3D technology employed in studies on neonates and children and uses an electromagnetic location device to register transducer position during data acquisition. As it derives from a complex assembly of sequentially acquired two-dimensional images this technique has important limitations due to fetal movement artifacts and difficulties in cardiac gating. Despite good three-dimensional appreciation of some anatomical detail, this often results in inadequate image quality when compared with 2D echocardiography. Potential advantages of 3D fetal echocardiography include the following: the ability to slice the acquired 3D volume data into an infinite number of two-dimensional cross-sections, and the ability to reconstruct unique three-dimensional views not seen with two-dimensional imaging. However, considering the limitations and the time needed for 3D image processing its practical clinical relevance in the antenatal situation is still not clear. Ongoing research should be aimed toward improved image resolution and applying this new technique to more immediate "on-line" analysis of 3D reconstructions. PMID- 11474998 TI - [27th annual meeting of the Society for Neonatology and Pediatric Intensive Care. Lubeck, 21-23 June 2001. Abstracts]. PMID- 11474999 TI - Treatment of watermelon stomach (GAVE syndrome) by means of endoscopic argon plasma coagulation (APC): long-term outcome. AB - We report on 17 patients with GAVE-syndrome (gastric antral vascular ectasia) treated by means of endoscopic argon plasma coagulation (APC). 16 of 17 patients presented with iron deficiency anemia; transfusion-dependent anemia was noted in 11 patients (65%). Resolution of the gastric angiectasia could be achieved in all patients by endoscopic APC after 1-4 treatment sessions. Endoscopic follow-up revealed recurrence of GAVE in 5 patients (requiring further treatment sessions). Mean pretreatment hemoglobin level of 78 g/l improved to 115 g/l after treatment. Only one patient needed post-treatment transfusions; she had refused further endoscopy. The mean follow-up was 30.4 months (range 1-65). In one case circumferential scarring of the antrum led to asymptomatic stenosis 6 months after APC; at the same time early recurrence of extensive angiectasia occurred. Billroth I resection was performed. No other complications were observed. Our results show that argon plasma coagulation is an effective and safe treatment for gastrointestinal blood loss due to GAVE syndrome (watermelon stomach). Control endoscopies are indicated in order to recognize and treat recurrence of angiectasia on time. PMID- 11474996 TI - [Clinical study of assessment of trisomy 21--precise risk evaluation in the first trimester of pregnancy]. AB - BACKGROUND: In the past a non invasive risk analysis for detecting specific chromosomal aberrations was only possible from week 15 of pregnancy. In this paper the practicability of first trimester screening is analysed. MATERIAL AND METHODS: Blood samples were taken from 1000 pregnant women before a invasive prenatal diagnosis was performed. Total hCG, free beta-hCG and PAPP-A (pregnancy associated plasma protein A) was analysed. These data were combined with complete cytogenetic and ultrasonographic (CRL and nuchal translucency--NT) data. RESULTS: In more than 90% of cases the NT was below 3 mm. Here the rate of normal karyotypes was 97.8%. In 61 cases a abnormal karyotype was found. Here in the most cases we found an elevated NT. Also in the most cases of trisomy 21 and 18 and in triploidies a characteristic ratio of hCG/free beta-hCG and PAPP-A was discovered. Combining NT and biochemical analysis, 85% of trisomies 21 could be discovered as a risk group. CONCLUSIONS: This study demonstrates the possibilities of first trimester screening with a high detection rate for specific chromosomal aberrations. DISCUSSION: First trimester screening should only be performed in specialised centers because determination of NT and risk analysis needs extensive experience. PMID- 11475000 TI - Lipid and DNA oxidative damage in experimentally induced hepatic porphyria in C57BL/10ScSn mice. AB - Patients with porphyria cutanea tarda (PCT) develop hepatocellular carcinoma as a late consequence. Pre-loading of C57BL/10ScSn mice with iron greatly sensitizes them to the induction of hepatic porphyria caused by hexachlorobenzene (HCB). HCB will also cause liver tumors in experimental animals. Elevated liver iron stores are implicated in the development of some human liver cancers in connection with its known catalytic role in generation of highly reactive activated oxygen species. The aim of this study was to determine the lipid and DNA oxidative damage in iron and HCB-induced porphyric mice. C57BL/10ScSn mice received i.p. injections of dextran sulfate (control), iron (Imferon) or combined iron and HCB. 6 weeks after treatment plasma ALT levels and hepatic free iron, porphyrin, lipid peroxides and 8-hydroxyguanosine (8-OHdG) levels were analyzed. Hepatic porphyrin level was significantly (p < 0.001) increased following combined iron/HCB treatment as compared to control mice. The level of lipid peroxides increased 9 fold (p = 0.001) and 35-fold (p < 0.001) after iron and iron/HCB treatment respectively, whereas the level of 8-OHdG was increased 2.5-fold (p = 0.002) and 7.5-fold (p < 0.001) after iron and iron/HCB treatment respectively as compared to control mice. The authors conclude that iron overload in conjugation with HCB induce lipid and DNA oxidative damage in C57BL/10ScSn mice. DNA oxidative damage may be important in the early events of hepatic carcinogenesis in experimental porphyria. PMID- 11475001 TI - [Gastrointestinal ultrasounds--possibilities for research and research funding]. AB - Sonography is nowadays the most used imaging procedure in gastroenterology. In contrast research in this field is little developed. However, a number of possibilities of research as regarding basic (technological) research as well in the context of patient oriented and outcome research can be proposed. If structure and quality conditions are adequate, a number of ways for funding sonographical research can be found which need to be used dependent on the category of research chosen. In particular outcome research should be considered for funding by public funding institutions (BMG/BMBF) and the insurance system. PMID- 11474997 TI - [Combined test in the first trimester of pregnancy in 3,316 unselected pregnant patients for diagnosing Down syndrome]. AB - The high detection rate (DR) for Down syndrome pregnancies which can be achieved by measuring fetal nuchal translucency (NT) early in pregnancy can be improved by combining it with placental hormones (PAPP-A, f beta hCG) and maternal age ("combined test"). In this study we wanted to assess the DR using the "combined test" in an unselected population of self-referred pregnant women at a false positive rate (FPR) of about 5%. MATERIALS AND METHODS: NT, PAPP-A, f beta hCG and maternal age were measured in all women with singleton pregnancies who booked for delivery in our hospital from 1.12.97 to 31.12.99 and who were between 10 and 13 completed weeks of gestation (crown-rump-length 35-70 mm). Calculation of the specific Down risk was done with the computer program Alpha, Version 5aa (Logical medical systems, London). RESULTS: A total of 3316 women were tested. Out of 10 Down pregnancies, which occurred in this period of time 9 could be detected with the test. 137 women had a positive test result but a non-Down pregnancy (FPR 4.1%). CONCLUSIONS: The combined test is an excellent test for the detection of Down syndrome pregnancies in a low-risk population. DISCUSSION: The constant increase in maternal age at the time of delivery can also lead to an improved DR if a simple age dependant protocol for Down-detection is used, but only at the price of a much higher number of amniocenteses and subsequent abortions. The DR for Down syndrome can be increased much more markedly using the "combined test" at a much lower FPR (approximately 5%). PMID- 11475003 TI - Mild thrombocytopenia as presenting symptom of type 1 Gauchers's disease. AB - A young woman was examined for a mild thrombocytopenia which was present for some months. No signs of bleeding had so far occurred. Physical examination was normal except for a moderately enlarged spleen. Laboratory investigations showed a low platelet count. There was no evidence of an autoimmune or hematologic disease. Bone narrow aspirate indicated Gaucher's-like cells raising the suspicion of Gaucher's disease. This was further supported by electron microscopic demonstration of Gaucher's bodies in crista biopsy specimens. However, the definitive diagnosis was obtained by verifying deficient lysosomal glucosylceramide-beta-D-glucosidase activity in peripheral blood leukocytes. Upon the absence of neurologic involvement the patient was typical for the adult-onset or type 1 form of Gaucher's disease. PMID- 11475002 TI - Gastric stromal tumor--a rare cause of an upper gastrointestinal bleeding. AB - Gastrointestinal stromal tumors (GISTs) are rare neoplasms arising from connective tissue elements of the gastrointestinal wall. They show a great heterogeneity with respect to their histogenetic, morphologic and prognostic characteristics. GISTs are known with myoid, neural or mixed features of differentiation. Clinical findings are gastrointestinal bleeding, abdominal pain and weight loss. We report on the case of a 50-year-old male patient who presented with melena and acute anemia (hemoglobin 10.5 g/dl). Esophagogastroduodenoscopy revealed a broad-based, centrally ulcerated polypoid formation of 3 cm in the gastric corpus as the cause of the upper gastrointestinal bleeding. Multiple endoscopic biopsies were negative for neoplastic changes. Because of no tendency of healing after triple eradication therapy of Helicobacter pylori and following proton pump inhibitor medication, the patient underwent distal gastrectomy with gastrojejunostomy. GIST of combined smooth muscle and neural type was diagnosed by histological and immunohistochemical examination. The features with increased mitotic activity and cellularity were those of a borderline stromal tumor. 6 months after surgery the patient is well with no signs of residual malignancy. This case demonstrates that rare stromal neoplasms have to be taken into account in the differential diagnosis of gastrointestinal tumors even if endoscopic biopsies are negative for neoplastic changes. Because of the uncertain biological behavior of the GISTs an early surgical intervention is recommended. PMID- 11475005 TI - [Colonoscopy is superior to colon double-contrast imaging in detection of recurrent polyps after polypectomy]. PMID- 11475004 TI - [Measuring quality of life in gastroenterology--concepts, instruments and problems]. AB - Health-related quality of life (HRQOL) is becoming an increasingly more important primary or secondary end point of clinical studies. Patients and their self help organizations demand a greater regard to their subjective experience of their disease and its treatment. Cost-effectiveness analyses measuring quality-adjusted life years are becoming more and more decisive in health politics. Therefore it is important for gastroenterologists to know the concepts and methods of HRQOL measurement. The present article discusses the definitions of (health-related) quality of life and reviews generic- and disease-specific quality of life instruments in gastroenterology including quality criteria. Rules for the adaptation of Anglo-American questionnaires into German-speaking countries are pointed out. Finally the limitations of the concepts of HRQUOL and its measurement are discussed. PMID- 11475008 TI - Protective effect of 7-nitroindazole against DSP-4 induced noradrenaline depletion in mouse hippocampus. AB - N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) is a selective noradrenaline (NA) uptake blocker, capable of inducing a long-lasting depletion of NA in some noradrenergic axon terminals originating from the locus coeruleus in rodents. Pretreatment with 7-nitroindazole, a fairly selective inhibitor of neuronal nitric oxide synthase in vivo, partially prevented DSP-4 induced NA depletion in mouse hippocampus measured seven days after the neurotoxic insult. Administration of L-arginine, the substrate of nitric oxide synthase, altered neither the NA depletion induced by DSP-4, nor the protective effect of 7-nitroindazole. Inhibition of nitric oxide synthesis by N(G)-nitro-L-arginine methyl ester did not attenuate the NA depleting effect of DSP-4. Thus, the contribution of neuronal nitric oxide synthase inhibition to the protective effect of 7 nitroindazole needs further studies. As 7-nitroindazole did not block NA uptake, this cannot play a part in the protective effect. The possible contribution of monoamine oxidase B enzyme inhibition by 7-nitroindazole to the protective effect is also discussed. PMID- 11475007 TI - Pineal indoles stimulate the gene expression of immunomodulating cytokines. AB - Male C57 mice received 10 consecutive daily intraperitoneal injections of melatonin, 5-methoxytryptamine or 5-methoxytryptophol (5mg/kg body weight). Control mice received the alcoholic saline vehicle. All mice were sacrificed 24 hours after the last injection. Following extraction of RNA from peritoneal exudate cells (PEC) and splenocytes, the level of gene expression was analyzed with the reverse transcription-polymerase chain reaction (RT-PCR). The results revealed that melatonin up-regulated the level of gene expression of transforming growth factor-beta (TGF-beta), macrophage-colony stimulating factor (M-CSF), tumor necrosis factor-alpha (TNF-alpha) and stem cell factor (SCF) in PEC, and the level of gene expression of interleukin-1beta (IL-1beta), M-CSF, TNF-alpha, interferon-gamma (IFN-gamma) and SCF in splenocytes. 5-Methoxytryptamine augmented the level of gene expression of TGF-beta, M-CSF and SCF in PEC, and the level of gene expression of IL-1beta, TNF-alpha, IFN-gamma, M-CSF and SCF in splenocytes. 5-Methoxytryptophol elevated the level of gene expression of TNF alpha, IL-1beta, TGF-beta and M-CSF in PEC, and the level of gene expression of inducible nitric oxide synthase (iNOS), IL-1beta, M-CSF, TNF-alpha, IFN-gamma and SCF in splenocytes. PMID- 11475006 TI - On the role of peripheral macrophages during active experimental allergic encephalomyelitis (EAE). AB - Experimental allergic encephalitis (EAE) is an experimental autoimmune inflammatory condition of the central nervous system (CNS) that serves as a disease model for multiple sclerosis (MS). The primary effector mechanisms of the immune system leading to tissue destruction during EAE remain still controversial. T-cells, microglia, and macrophages infiltrating the brain parenchyma are suggested to be involved. To clarify the role of these cells during disease Lewis rats were immunised with different immunisation protocols: Immunisation with myelin basic protein (MBP) in complete Freunds adjuvant (CFA) containing high dose of mycobacterial components induced severe disease, whereas immunisation with low dose of mycobacterial components induced only mild disease. Severely and mildly diseased animals were analysed with respect to infiltration of T-cells, macrophages and upregulation of MHC class II molecules on microglia in the brain. All immunised rats showed high T-cell infiltration accompanied by microglia activation. The degree of disease and the infiltration of macrophages varied with dose of adjuvant. Lowering the dose of adjuvant prevented the development of disease but also the influx of peripheral macrophages into the brain without affecting the peripheral T-cell response to the autoantigen. Thus, appearance of (autoreactive) T-cells in the brain and microglia activation were probably not sufficient for development of disease. It can be concluded that peripheral macrophages play an essential or even key role in the pathogenesis of active EAE. PMID- 11475009 TI - Scopolamine does not restore normal conditioned avoidance performance in raclopride-treated rats. AB - Several studies have shown antagonism by anticholinergics of antipsychotic induced suppression of conditioned avoidance behavior, as well as of catalepsy, in rats. These observations provide pharmacological evidence for these behaviors mediated via nigro-striatal dopaminergic projections, as well as known dopaminergic-cholinergic interactions in the neostriatum. The objective of the present study was to examine the quality of behavioral change produced by scopolamine (0.25-1.00 mgkg(-1) s.c.) on conditioned avoidance behavior, by itself, and in combination with raclopride (0.1 mgkg(-1) s.c.) in the rat. Adult male Wistar rats were trained to perform a conditioned avoidance response requiring a brightness discrimination. A two-way avoidance shuttle-box was used with the modification that there were two passages in the partition separating the two compartments of the shuttle-box. In order to make a correct avoidance in the response to white noise conditioned stimulus, the rat had to take background light into consideration. Correct passage under dim background conditions was to the left and, with increased background lights, to the right. A weak, intermittent, electric shock (approximately 0.2 mA) was used as unconditioned stimulus. Scopolamine by itself (0.25-1.00 mgkg(-1) s.c.) disrupted the visual discrimination without affecting avoidance performance. As expected, raclopride (0.1 mgkg(-1) s.c.) produced a suppression of conditioned avoidance behavior. A dose of 1.00 mgkg(-1) of scopolamine was needed to restore raclopride-induced suppression of conditioned avoidance behavior. Thus, restoration of the avoidance behavior by scopolamine treatment was not possible at doses that allow normal performance of the visual discrimination. It is concluded that anticholinergics do not restore normal behavior after neuroleptic-induced suppression of conditioned avoidance behavior. PMID- 11475011 TI - A study of alpha-adrenoceptor gene polymorphisms and Alzheimer disease. AB - There exists considerable evidence implicating abnormalities of the alpha (alpha) adrenergic system in the development of Alzheimer disease (AD). We propose to investigate potential correlations between the presence or otherwise of alpha adrenoceptor polymorphisms and the presence of AD. We studied the polymorphisms of the alpha1a- and the alpha2a-adrenoceptor genes in 142 AD patients and 98 normal controls. The result demonstrated that none of the alpha2a-adrenoceptor genotypes was associated with increased susceptibility to AD. However, there was a trend that the frequency of the C allele of the alpha1a-adrenoceptor was elevated and an excess of the CC genotype (90.1%) was found in the subjects with AD in comparison with the controls (78.6%). This association was unrelated to the apolipoprotein E genotypes. The hypothesis that the alpha1a-adrenoceptor gene may be implicated in the pathogenesis of AD may deserve further study. PMID- 11475012 TI - Tacrine and rate of progression in Alzheimer's disease--relation to ApoE allele genotype. AB - Today, cognitive impairment can be successfully treated with acetylcholine esterase inhibitors (AChE-I) in many, but not all, patients with Alzheimer's disease (AD). To investigate the relation between tacrine treatment, inheritance of ApoE epsilon4 alleles, and rate of progression, the differences in MMSE and CIBIC scores (efficacy parameters) after 6 and 12 months of tacrine (an AChE-I) treatment were investigated in 145 AD patients. Of these, 84 were ApoE epsilon4 positive (ApoE4) and 61 were ApoE epsilon4-negative (ApoE2-3). No differences were found after 6 months of treatment, but after 12 months the CIBIC scores revealed that the ApoE4 patients had declined more than the ApoE2-3 patients (p < 0.05). No differences were found for the last 6 months of treatment. The results primarily suggest a faster rate of decline in the ApoE4 AD compared to the ApoE2 3, but may also reflect that ApoE epsilon4 genotype inheritance is a negative predictor of treatment effect of tacrine in AD patients. PMID- 11475010 TI - Decreased densities of dopamine and serotonin transporters and of vesicular monoamine transporter 2 in severely kainic acid lesioned subregions of the striatum. AB - Fifteen days after a striatal kainic acid (KA) injection, we have examined presynaptic modifications of dopamine and serotonin terminals in the striatum through (i) autoradiographic labeling of dopamine, serotonin and vesicular monoamine transporters respectively with 3H-mazindol, 3H-citalopram and 3H dihydrotetrabenazine, and (ii) determination of the contents in dopamine, serotonin and their metabolites. Acetylcholinesterase histochemical labeling enabled the definition of severely and moderately KA-lesioned subregions within the striatum. A significant decrease of the three transporters labeling density was observed only in the severely lesioned subregions. The strong decrease in serotonin transporter labeling revealed here has not been described until now. Besides, the striatal contents of homovanillic acid (dopamine metabolite) and 5 hydroxyindolacetic acid (serotonin metabolite) were significantly increased in the lesioned striatum. The whole data evidence an incomplete sparing of dopamine and serotonin terminals in the striatum 15 days after a KA injection, especially in the areas where the degeneration of postsynaptic neurons was the most extensive. PMID- 11475014 TI - Neurotoxicity of 24-hydroxycholesterol, an important cholesterol elimination product of the brain, may be prevented by vitamin E and estradiol-17beta. AB - 24-Hydroxycholesterol, the main cholesterol elimination product of the brain is increased in serum of Alzheimer patients. This oxysterol behaves neurotoxic towards the human neuroblastoma cell line, SH-SY5Y. Here we demonstrate, that 24 hydroxycholesterol-induced neurotoxicity in differentiated SH-SY5Y cells was due to apoptosis, as indicated by DNA-fragmentation, caspase-3 activation and a decrease of the mitochondrial membrane potential. Free radicals were generated, resulting in the death of 75% of the cells within 48h; neurotoxicity in differentiated SH-SY5Y cells was partially prevented by physiological concentrations of vitamin E (50-100 microM) in that 75% of the cells survived. Physiological concentrations of estradiol-17beta (1-100nM) elicited a protective effect in differentiated cells, which was not significant; however, in undifferentiated cells a significant protection was noted by this steroid hormone. Vitamin C and melatonin did not prevent 24-hydroxycholesterol-induced neurotoxicity. These in vitro data support the in vivo observed beneficial effects reported as circumstantial evidence of vitamin E and estradiol-17beta treatment in the prevention and therapy of neurodegenerative disease. PMID- 11475013 TI - Antiapoptotic effects of the peptidergic drug cerebrolysin on primary cultures of embryonic chick cortical neurons. AB - Cerebrolysin (EBEWE Arzneimittel, Austria, Europe) is a widely used drug relieving the symptoms of a variety of neurological disorders, particularly of neurodegenerative dementia of the Alzheimer's type. It consists of approximately 25% of low molecular weight peptides (<10k DA) and a mixture of approximately 75% free amino acids, this being based on the total nitrogen content. In this study we used a low serum (2% serum supplement) cell stress in-vitro model to assess drug effectiveness on neuronal viability and programmed cell death (PCD). In this in-vitro model the type of cell death was previously shown to be primarly apoptotic, which was verified by DNA-laddering and TUNEL-staining. For evaluation of neuronal viability a MTT-reduction assay was performed after 4 DIV and 8 DIV and the percentage of apoptotic neurons was determined by bis-benzimide staining of nuclear chromatin. To differentiate between possible effects of the free amino acids and the peptide fraction of Cerebrolysin an artificial amino acid mixture (AA-mix) was used as a control. Cerebrolysin, the AA-mix and 10% foetal calf serum (FCS) caused a similar increase in viability after 4 DIV, whereas the effects of the growth factors BDNF and FGF-2 were less pronounced. After 8 DIV Cerebrolysin, but not the AA-mix, was able to ameliorate neuronal viability, which could reflect a neuro-protective effect or an increased activity of the mitochondrial dehydrogenase measured in a MTT-reduction assay. The percentage of cells showing apoptotic chromatin changes was significantly reduced (p < 0.01) in cultures treated with Cerebrolysin, whereas the AA-mix failed to decrease the percentage of cells showing apoptotic chromatin changes. These findings ascertain an anti-apoptotic effect of the peptide fraction of Cerebrolysin and reveal a transient viability promoting effect of the amino acid fraction, which is most likely due to improved nutritional supply. PMID- 11475015 TI - An association of the polymorphic repeat of tetranucleotide (TCAT) in the first intron of the human tyrosine hydroxylase gene with schizophrenia in a Japanese sample. AB - Tyrosine hydroxylase (TH) is the rate limiting enzyme in the synthesis of dopamine and norepinephrine. A polymorphic repeat of the tetranucleotide (TCAT) in the first intron of the TH gene may behave as a regulatory element for the gene expression. Allelic fragments of the tandem repeat were typed by a PCR-based process with a pair of primers specific for the polymorphic sequence. The association between the polymorphism and schizophrenia was examined in a Japanese sample. There was a statistically significant association between the polymorphism and schizophrenia in females (chi2 = 26.018, p = 0.010), but not in males (chi2 = 15.995, p = 0.305). The genotype heterozygous for the TH9 and TH6 was significantly decreased in female schizophrenics (chi2 = 5.125, p = 0.0236). These results suggest that TH could be considered as a minor gene contributing to the susceptibility of Japanese female schizophrenia. PMID- 11475016 TI - [Three patients with massive pulmonary embolism]. AB - Three patients presenting with massive venous pulmonary thrombo-embolism are described, who have been selected from a series of 22 patients treated with thrombolysis during a 6-year period. A 23-year-old female presented with tachycardia and dyspnoea. She had pulmonary angiography following scintigraphy with a perfusion deficit of more than 60%. Thrombolysis resulted in open blood vessels and a disappearance of the complaints. A 51-year-old woman presented with profound hypoxemia, probably due to a patent foramen ovale, with shunting and tachycardia. Perfusion defects on scintigraphy combined with a normal chest radiograph in the absence of pre-existent pulmonary disease established the diagnosis. She responded favourably to intravenous streptokinase. The third patient was an 80-year-old woman with hypertension. She developed dyspnoea, tachycardia and shock following immobilisation due to a fractured hip. Despite an initial improvement on streptokinase, she deteriorated and died from right-sided heart failure. The diagnostic tests should be limited and aimed at ruling out left-sided heart failure and pericardial tamponade. Echocardiography is often diagnostic in these patients. Thrombolysis may be life saving but there are no randomised trials to prove that survival rate is indeed better compared to heparin therapy. Streptokinase is less expensive than alteplase and there is no evidence from trials to suggest that it is inferior to more expensive thrombolytics such as alteplase or urokinase. PMID- 11475017 TI - [Netherlands Health Council report:'Risks associated with folic acid fortification': those who don't risk, don't win]. AB - Because the advice to take folic acid supplements is insufficiently followed by women who wish to become pregnant, a committee of the Health Council of the Netherlands investigated the possible risks associated with increasing folic acid intake through fortification of food products. The committee concludes that the risk of inducing or exacerbating neurological abnormalities by increasing the intake of folic acid in people with a vitamin B12 deficiency cannot be excluded. Therefore the fortification of food products should, for the time being, be limited to those foods which are especially aimed at women who wish to become pregnant. It is unlikely, however, that such a restricted food fortification program will lead to a decrease in the incidence of neural tube defects. Like the use of folic acid supplements, the purchase of special food products presupposes that the individual woman will undertake a positive action. Moreover, most women do not want their shopping to reveal their wish to have a child, and the need to cook their own meals will meet practical difficulties. The hypothetical and avoidable side-effects of food fortification have to be balanced against the certain benefit of preventing neural tube defects in unplanned pregnancies and pregnancies in women unaware of the benefits of folic acid supplementation. A revision of the report is recommended. PMID- 11475018 TI - [Virus quantification in peripheral blood as a reflection of the clinical course]. AB - Quantitative determination of viral nucleic acid in peripheral blood appears to be important for nearly all viral infections with a chronic and reactivating course, as this provides relevant information on the clinical consequences, the level of infectivity and the response to antiviral treatment or other interventions. The practical application of this principle in clinical virology has benefited from the availability of new techniques, particularly those in which amplification of viral nucleic acid can be followed in real time. In addition, the potential value of quantitative virology was clearly demonstrated by the experience gained from viral load monitoring in HIV-infected patients. This approach will probably be widely applicable in clinical microbiology in general. PMID- 11475021 TI - [Increased prevalence of overweight and obesity in Dutch children, and the detection of overweight and obesity using international criteria and new reference diagrams]. AB - OBJECTIVE: To determine the prevalence of overweight and obesity in Dutch children in 1980 and 1997 according to international criteria, and to design new reference diagrams for overweight and obesity in children. DESIGN: Descriptive. METHOD: The prevalence of overweight and obesity, based on height and weight data from the Fourth Dutch Growth Study (1997), was determined according to international criteria for age and sex. RESULTS: In 1997, the prevalence of overweight and obesity increased in both boys and girls compared to 1980: in 1997, the prevalence of overweight ranged between 7.1 and 15.5% for boys, and between 8.2 and 16.1% for girls. Both the prevalence of overweight and obesity was higher in girls than in boys. CONCLUSION: By applying the international criteria for overweight and obesity and reference growth diagrams based upon the 1997 Dutch growth study, the prevention and detection of overweight and obesity has to be implemented with vigour in Dutch youth health care. PMID- 11475020 TI - [Diagnostic image (45). Ochronosis]. AB - In a 70-year-old woman, in whom ochronosis (alkaptonuria) was diagnosed at the age of 54, bluish discolouration of the cartilage of the ears was observed. PMID- 11475019 TI - [Abdominal obesity: metabolic complications and consequences for the liver]. AB - Obesity is associated with a number of metabolic and haemodynamic risk factors for cardiovascular disease and type 2 diabetes mellitus. This risk depends on a complex of metabolic and haemodynamic consequences of (visceral) fat accumulation, which probably results from the continuous delivery of fatty acids to the liver via the portal vein. Hypertriglyceridaemia, hyperinsulinaemia, hypertension, insulin resistance and increased hepatic glucose production are all independent risk factors for atherosclerosis. Their combination increases the risk of cardiovascular disease considerably. Triglyceride storage in hepatocytes is another consequence of increased fatty acid supply to the liver. Until recently, hepatic steatosis was considered a harmless condition secondary to obesity or alcoholism. However, it may lead to non-alcoholic hepatic steatosis, which predisposes to liver fibrosis and even cirrhosis. PMID- 11475023 TI - [Evaluation of the Dutch Medical Treatment Act (WGBO)]. PMID- 11475022 TI - [Misinterpretation of nerve damage to the hand and forearm due to variations in the motor innervation]. AB - A 21-year-old man served his ulnar nerve at the level of the left elbow. After nerve suturing only a slight loss of strength in two ulnar intrinsic hand muscles was found. Electromyographic examination revealed the crossing of some motor nerve fibres in the forearm from the median nerve to the ulnar nerve, innervating most of the first dorsal interosseous and the abductor digiti minimi muscle. One and a half years later there was still a loss of strength in the other ulnar muscles and a reduced sensitivity of the ulnar area, without a loss of function. Variations in the motor nerve supply of the hand, in particular connections between the median nerve and ulnar nerve in the forearm and hand are common. A complete transection of the ulnar nerve without complete paralysis of the ulnar intrinsic hand muscles can occur. If these variations are not considered, such a transection can be overlooked. PMID- 11475025 TI - [Hydatid cyst of the adductor muscles. MRI aspect, Apropos of 1 case]. AB - Muscular involvement is rare, representing less than 1% of hydrated cyst localisation's. Clinical symptoms are insidious and non-specific causing a frequent delay in diagnosis. We report the observation of a 50 year-old woman consulting for inguinal tumefaction with radiological exploration suggestive of hydrated cyst of adductors muscles. Magnetic resonance imaging is helpful in diagnosis, since it reveals a very suggestive aspect. Treatment is based on surgery, which cures patients but incurs a low risk of local relapse. PMID- 11475026 TI - [Kawasaki syndrome: apropos of 2 cases observed in Brazzaville]. AB - The authors report for first time in Congo two cases of Kawasaki syndrome. One case from a five years boy carried out a typical form of an adeno-mucocutaneous syndrome; the other from a six month little girl was an incomplete form. In both cases, the immediate development has been excellent. The scarceness of this syndrome among the blacks and the diagnostic difficulties have been the background of the interest of this work. PMID- 11475024 TI - [Ambulatory management of sickle cell disease: evaluation of the first year follow up of patients in the pediatric department of Lome (Togo)]. AB - BACKGROUND: This study is an evaluation of the first year ambulatory follow up of patients from the sickle-cell care centre of the paediatric ward of the teaching hospital in Lome-Tokoin. PATIENTS AND METHODS: Togo is situated in the epicentre of the Benin haplotype. A total of 132 patients (109 SS, 22 SC and 1 S beta zero thal) followed up during one year from their admission date (period of 1st January 1996 to 31st December 1997). 132 patients were included in the study. RESULTS: The patients' age varied, for the majority, between 2 months and 15 years, but a few adults (15%) were included in the study. Information was collected from the hospital files and health cards, which unfortunately did not have specific entrees for sickle cell disease. Clinical features revealed that the frequency of tooth decay and chronic persistent splenomegaly was low when compared to the rates in central Africa (Bantu haplotype). Laboratory findings lead to the conclusion that some analysis are relevant such as the dosage of the G6PD activity (24.1% of patients were deficient), parasitologic analysis of faeces (positive in 22.5%), retinal fluoro-angiography (32.2% of ocular lesions), and cardiologic check-up. On the other hand, scanning of biliary tracts and systematic X-rays of the hips seems to be secondary. Some positive results were noticed by the scanning of biliary tracts without any therapeutic decisions in non-symptomatic patients; no case of osteonecrosis was detected by the X-rays. The mean haemoglobin level was 7.4 +/- 1.4 g/dl for the SS and 10.7 +/- 2.4 g/dl for the SC. The mean MCV were 91.3 +/- 10.1 fl and 82.1 +/- 7.7 fl, respectively. Specific vaccinations were not well performed because of their high cost. CONCLUSION: In order to carry on and improve the ambulatory management of patients with sickle cell disease, it is important in low income countries, such as Togo, to target the necessary laboratory tests for an initial and annual check up. Solidarity networks for patients should be promoted and effective involvement of the health authorities ensured. PMID- 11475030 TI - [Sickle cell disease and thalassemia: examples of a Darwinian approach to disease]. PMID- 11475027 TI - [Behavioral disorders after treatment with isoniazid]. AB - Isoniazid, efficient antituberculosis drug, can provoke neuropsychiatric manifestations at certain patients. Two mechanisms of action for isoniazid related psychosis are kept by the majority of authors: pyridoxin deficiency isoniazid toxicity, molecule near chemically to iproniazid, powerful IMAO (monoamine oxydase inhibitor). The predisposing factors are: slow acetylator, diabetes, hepatic insufficiency, old age, alcoholism. Family and personal history of mental illness are also predisposing factors. We report here the observation of a 53-year-old man presented with psychotic symptoms suspected to be relation with isoniazid. The favorable evolution after the definitive stop of isoniazid therapy is in favor of this hypothesis. Although rare, the neuropsychiatric symptoms during tuberculosis treatment by isoniazid, the possibility of iatrogenic etiology must be evoked. Isoniazid, antituberculosis very used in Africa, is mentioned by several authors like the cause of psychiatric disorders. Pyridoxin deficiency seems to play a role of trigger. Supplementation with vitamin B6 during the treatment by isoniazid must be therefore systematic to warm these unrests. PMID- 11475031 TI - [Physiopathological basis of sickle cell disease, management and current therapeutics]. AB - Physiopathological knowledge in the area of sickle cell disease underwent a first major development three decades ago, some 15 years after the discovery of the molecular nature of the disease. Several physiopathological areas can be distinguished: physiopathology of polymerisation which has been successfully approached by fundamental biophysicists and biochemists alike; physiopathology of the red blood cell; more recently, the characterisation of red blood sickle cells by extra-hemoglobin aspects: the red blood cell is not a passive recipient for hemoglobin but carries out a series of functions which can contribute to physiopathology; the phenomenon of vaso-obstruction is still poorly understood, whereas clinical secondary manifestations of the disease, the physiopathology of infectious risk and severe anaemic manifestations have been well understood. The treatment of sickle cell patients continues to evolve and progress. There are certain characteristics which remain, however, permanent: there is continual interaction between preventive and therapeutic measures; treatment must be both medical and social, given the impact of the disease of the individual and his family or vice-versa; treatment is centred around the hospital with more than ever the necessity to ensure excellence in internal medicine or paediatrics, intensive care and transfusion. Finally, the distinction among three periods for the natural history of the disease remain: before 7 years of age, from 7 years to adult age, adulthood. An integrated, pilot centre for treating patients with sickle cell disease is of crucial importance and can contribute to therapeutic innovations in the area. PMID- 11475029 TI - [Resurgence of the plague in the Ikongo district of Madagascar in 1998. 2. Reservoirs and vectors implicated]. AB - Our survey of mammals and fleas arose as a result of an outbreak of bubonic plague at an usually low altitude in the Ikongo district (Madagascar), while a previous study had found anti-F1 antibodies in an endemic hedgehog. Animals were sampled with live traps in two hamlets (Antanambao-Vohidrotra, 540 m alt. and Ambalagoavy, 265 m alt.) and with pitfall traps in a neighbouring forest (750 m alt.). Rat fleas were collected by brushing the fur and free-living fleas by use of light traps. The introduced shrew Suncus murinus was found only in the village of Ambalagoavy while the black rat (Rattus rattus) was found in all three sites and the only seropositive rat was caught at Antanambao-Vohidrotra. In contrast, among the Tenrecidae (endemic shrews and hedgehogs) found in the forest near the first village, four animals were found seropositive for anti-F1 antibodies. One of them was carrying the endemic flea Paractenopsyllus pauliani, not yet reported as a vector of plague. The endemic vector of plague, Synopsyllus fonquerniei, was found only in the first village of Antanambao-Vohidrotra, and the cosmopolite flea Xenopsylla cheopis only in Ambalagoavy. Although no Yersinia pestis could be isolated and no F1-antigen could be detected in these animals, we found evidence of the recent transmission of plague in Antanambao-Vohidrotra and the nearby forest, but not in Ambalagoavy. These data corroborate with the sylvatic plague cycle hypothesis in Madagascar and its involvement in the outcome of the bubonic plague outbreak in this district. PMID- 11475028 TI - [Resurgence of the plague in the Ikongo district of Madagascar in 1998. 1. Epidemiological aspects in the human population]. AB - Between the 20th October and the 18th November 1998, an outbreak of bubonic plague was declared in a hamlet in the Ikongo district of Madagascar. We conducted an epidemiological survey because of the re-emergence of the disease in this area (the last cases had been notified in 1965) and because of the low altitude compared to the classical Malagasy foci. The outbreak had been preceded by an important rat epizootics during September. A total of 21 cases were registered with an attack rate of 16.7% (21/126) and a lethality rate of 33% (7/21). The disease was more prevalent in males (66% of cases) and children aged < 15 years, as observed in general throughout the country. The anti-F1 seroprevalence among the contact population was 13.5% (13/96), probably attributable to subclinical infection by Yersinia pestis. No rodent was trapped during the survey, but an endemic hedgehog (Tenrec ecaudatus) was highly seropositive, suggesting a recent transmission of the plague bacillus among this species. The small mammals and vectors possibly involved in these new foci were investigated in May 1999. PMID- 11475032 TI - [Sickle cell disease: laboratory and hemoglobin study]. AB - The diagnosis of any sickle cell disease syndrome is based on the unambiguous identification of HbS. Electrophoretic tests are usually the first to be performed. A much better resolution is obtained with isoelectricfocusing than with the more conventional cellulose acetate electrophoresis at alkaline pH. In some laboratories the first test is cation exchange HPLC. The diagnosis of HbS should never be accepted if not confirmed by a second test, more specific of this Hb such as the solubility test or electrophoresis on agar in citrate buffer. The laboratory should also evaluate other factors interacting with HbS, such as HbF level, sickle cell restriction haplotype, association with alpha-thalassemias. It should also evaluate other cellular factors and, in case of symptomatic heterozygous patients, help to understand of the underlying mechanisms. PMID- 11475035 TI - [Beta-thalassemia: clinical manifestations]. AB - Monogenic diseases are very unevenly distributed throughout the world and beta thalassemies are due chiefly to a large number of point mutations of the beta globine gene. The thalassemia trait (heterozygous thalassemia) can be asymptomatic and diagnosis is established by demonstration of an increased proportion of Hb A2. In the homozygous state (thalassemia major) hypochromic anemia is extremely severe because erythropoiesis is largely ineffective. Regular transfusion is necessary to prevent early death and transfusion therapy is usually initiated in the first year of life after biological diagnosis. Iron chelation is now capable of preventing transfusional haemachromatosis responsible for late mortality. 10% only of patients with homozygous beta-thalassemia have a syndrome of intermediate haematologic severity (thalassemia intermedia). Hb S- beta-thalassemia disease is characterised by a clinical course that resembles more that of Sickle Cell disease than to the thalassemia syndromes. PMID- 11475034 TI - [Treatment of major sickle cell syndromes. The point of view of the internist]. AB - Major sickle cell syndromes constitute at present the most important risk of genetic disease in the wider Paris area. Sickle cell patients are exposed to vaso occlusive attacks and to chronic visceral complications of ischemic origin capable of affecting all organs. Acute vaso-occlusive complications represent the first cause of hospitalisation and the main cause of death for the adult patient. Functional asplenia also occurs with a risk of infection through encapsuled germs. The risk of infection must always be taken into account by clinicians treating adults even if serious infectious accidents are less frequent in adults than in children. Sickle cell disease is a public health problem. Better knowledge of acute vaso-occlusive complications and ways of treating them rapidly by simple symptomatic measures mean that the mortality rate and the risk of serious functional sequelae can be decreased. Such measures allow sickle cell patients to enjoy a life expectancy beyond 50 years. PMID- 11475036 TI - [Therapeuetic management of patients with thalassemia major]. AB - In industrialised countries, the use of regular blood transfusions and of chelation therapy with Deferoxamine (DFO) has led to the transformation of thalassemia major from a fatal disease in early childhood to a chronic illness associated with prolonged survival. Transfusion regimens maintaining pretransfusion hemoglobin > 9-10 g/dl are effective in suppressing erythroid marrow expansion. Long term DFO therapy using subcutaneous infusions at least 4-6 d a week have clearly demonstrated major effects on iron overload complications. DFO treatment reduces excessive iron and prevents cardiac, hepatic and endocrine diseases. Nonetheless, compliance is difficult for many patients and the cost of DFO limits its use in developing countries. The only oral iron chelating agent that has been investigated extensively is Deferiprone (L1). In France, this oral agent can be administered in patients experiencing toxic side effects under DFO treatment. Since 1981 more than 1500 bone-marrow transplants have been performed word-wide, mostly in Italy. Allogenic BMT is currently able to cure 85% of thalassemic children with an available HLA matched sibling donor. PMID- 11475038 TI - A decision-making model for health promotion advocacy: the case for advocacy of drunk driving control measures. AB - This paper presents a decision-making model which can help public health professionals justify their decision to advocate for a particular intervention. The model is demonstrated by a case study related to advocacy of Random Breath Testing (RBT). For the purpose of this paper advocacy is a "catch-all word for the set of skills used to create a shift in public opinion and mobilise the necessary resources and forces to support an issue, policy, or constituency..." (Wallack, Dorfman, Jernigan & Themba, 1994). PMID- 11475037 TI - [Prevention of hemoglobinopathies in non-endemic countries]. AB - Although hemoglobinopathies are primarily found in Africa, India, SouthEast Asia and the Mediterranean area, their distribution is becoming worldwide due to increased migration. Unlike other genetic diseases, carriers can be detected by simple and cost-effective means. Prenatal hemoglobinopathy screening is possible and direct prenatal diagnosis can be offered to couples at risk of giving birth to a child affected by a major defect of the beta-globin chain. Several hemoglobinopathy screening programmes have been organised in various countries of Northern Europe and have been effective in identifying couples at risk. PMID- 11475033 TI - [Sickle cell disease: from childhood to adolescence. Management in 2001]. AB - Sickle cell anaemia is a chronic disease which evolves on the background of a basal state punctuated by critical periods, often with complications requiring overall treatment. The management of both periods must be well integrated into the treatment programme. In order to treat occlusive crisis, medical staff must be well trained and equipped for the complicated clinical diagnoses involved in drepanocytosis: bone infarction or osteomyelitis, pneumonia or acute chest syndrome, multifocal crisis or severe infectious disease. Neonatal diagnosis is the best way for early treatment by preventing severe pneumococcal diseases, ensuring sufficient hydration and appropriate surveillance, as well as providing family education. Much progress has been made over the last decade in the early treatment of retinitis, biliary lithiasis, and aseptic hip necrosis. It is also important to ensure accurate prescriptions for hydroxyurea. Specialised centres for drepanocytosis treatment are now a growing necessity and should be encouraged in order to provide treatment for dispersed patients against a relatively little known disease. PMID- 11475041 TI - Developing a partnership for HIV primary prevention for men at high risk for HIV infection in rural communities. PMID- 11475040 TI - The role of advocacy in promoting health. PMID- 11475039 TI - Peer education. PMID- 11475042 TI - International governance and reform of the World Trade Organization. PMID- 11475046 TI - Phylogenetic autocorrelation under distinct evolutionary processes. AB - I show how phylogenetic correlograms track distinct microevolutionary processes and can be used as empirical descriptors of the relationship between interspecific covariance (V(B)) and time since divergence (t). Data were simulated under models of gradual and speciational change, using increasing levels of stabilizing selection in a stochastic Ornstein-Uhlenbeck (O-U) process, on a phylogeny of 42 species. For each simulated dataset, correlograms were constructed using Moran's I coefficients estimated at five time slices, established at constant intervals. The correlograms generated under different evolutionary models differ significantly according to F-values derived from analysis of variance comparing Moran's I at each time slice and based on Wilks' lambda from multivariate analysis of variance comparing their overall profiles in a two-way design. Under Brownian motion or with small restraining forces in the O U process, correlograms were better fit by a linear model. However, increasing restraining forces in the O-U process cause a lack of linear fit, and correlograms are better described by exponential models. These patterns are better fit for gradual than for speciational modes of change. Correlograms can be used as a diagnostic method and to describe the V(B)/t relationship before using methods to analyze correlated evolution that assume (or perform statistically better when) this relationship is linear. PMID- 11475047 TI - Genetic trends in a population evolving antibiotic resistance. AB - The evolution of antibiotic resistance provides a well-documented, rapid, and recent example of a selection driven process that has occurred in many bacterial species. An exhaustive collection of Moraxella catarrhalis that spans a transition to chromosomally encoded penicillin resistance was used to analyze genetic changes accompanying the transition. The population was characterized by high haplotypic diversity with 148 distinct haplotypes among 372 isolates tested at three genomic regions. The power of a temporally stratified sample from a single population was highlighted by the finding of high genetic diversity throughout the transition to resistance, population numbers that remained high over time, and no evidence of departures from neutrality in the allele frequency spectra throughout the transition. The direct temporal analysis documented the persistence, antibiotic status, and haplotypic identity of strains undergoing apparent clonal expansions. Several haplotypes that were beta-lactamase nonproducers in early samples converted to producers in later years. Maintenance of genetic diversity and haplotype conversions from sensitive to resistant supported the hypothesis that penicillin resistance determinants spread to a diverse array of strains via horizontal exchange. Genetic differentiation between sample years, estimated by F(ST), was increasing at a rate that could cause complete haplotype turnover in less than 150 years. Widespread linkage disequilibrium among sites within one locus (copB) suggested recent mutation followed by clonal expansion. Nonrandom associations between haplotypes and resistance phenotypes provided further evidence of clonal expansion for some haplotypes. Nevertheless, the population structure was far from clonal as evidenced by a relatively low frequency of disequilibria both within sites at a second locus (M46) as well as between loci. The haplotype-antibiotic resistance association that was accompanied by gradual haplotype turnover is consistent with a hypothesis of genetic drift at marker loci with directional selection at the resistance locus. PMID- 11475044 TI - The evolution of postzygotic isolation: accumulating Dobzhansky-Muller incompatibilities. AB - Hybrid sterility and inviability often result from the accumulation of substitutions that, while functional on their normal genetic backgrounds, cause a loss of fitness when brought together in hybrids. Previous theory has shown that such Dobzhansky-Muller incompatibilities should accumulate at least as fast as the square of the number of substitutions separating two species, the so-called snowball effect. Here we explicitly describe the stochastic accumulation of these incompatibilities as a function of time. The accumulation of these incompatibilities involves three levels of stochasticity: (1) the number of substitutions separating two allopatric lineages at a given time; (2) the number of incompatibilities resulting from these substitutions; and (3) the fitness effects of individual incompatibilities. Previous analyses ignored the stochasticity of molecular evolution (level 1) as well as that due to the variable effects of incompatibilities (level 3). Here we approximate the full stochastic process characterizing the accumulation of hybrid incompatibilities between pairs of loci. We derive the distribution of the number of incompatibilities as a function of divergence time between allopatric taxa as well as the distribution of waiting times to speciation by postzygotic isolation. We provide simple approximations for the mean and variance of these waiting times. These results let us estimate. albeit crudely, the probability, p, that two diverged sites from different species will contribute to hybrid sterility or inviability. Our analyses of data from Drosophila and Bombina suggest that p is generally very small, on the order of 10(-6) or less. PMID- 11475043 TI - Perspective: From mutants to mechanisms? Assessing the candidate gene paradigm in evolutionary biology. AB - The generation of mutants in model organisms by geneticists and developmental biologists over the last century has occasionally produced phenotypes that are startlingly reminiscent of those seen in other species. Such extreme mutations have generally been dismissed by evolutionary geneticists since the "modern synthesis" as irrelevant to adaptation and speciation. But only in recent years has information on the molecular bases of mutant phenotypes become widely available, and thus work on testing the relevance of such extreme mutations to the generation of phylogenetic diversity has just begun. Here we evaluate whether evolutionary mimics are, in fact, useful for pinpointing the genetic differences that distinguish morphological variants generated during evolution. Examples come from both plants and animals, and range from intraspecific to interordinal taxonomic ranges. The use of mutationally defined candidate genes to predict evolutionary mechanisms has so far been most fruitful in explaining intraspecific variants, where it has been effective in both plants and animals. In several cases these efforts were facilitated or supported by parallel results from quantitative trait loci studies, in which natural alleles controlling continuous variation in developmental model organisms were mapped to mutationally defined genes. However, despite these successes the approach's utility seems to rapidly decay as a function of phylogenetic distance. This suggests that the divergence of developmental genetic systems is great even in closely related organisms and may become intractable at larger distances. We discuss this result in the context of what it teaches us about development, the future prospects of the candidate gene approach, and the historical debate over process in micro- and macroevolution. PMID- 11475045 TI - How closely correlated are molecular and quantitative measures of genetic variation? A meta-analysis. AB - The ability of populations to undergo adaptive evolution depends on the presence of quantitative genetic variation for ecologically important traits. Although molecular measures are widely used as surrogates for quantitative genetic variation, there is controversy about the strength of the relationship between the two. To resolve this issue, we carried out a meta-analysis based on 71 datasets. The mean correlation between molecular and quantitative measures of genetic variation was weak (r = 0.217). Furthermore, there was no significant relationship between the two measures for life-history traits (r = -0.11) or for the quantitative measure generally considered as the best indicator of adaptive potential, heritability (r = -0.08). Consequently, molecular measures of genetic diversity have only a very limited ability to predict quantitative genetic variability. When information about a population's short-term evolutionary potential or estimates of local adaptation and population divergence are required, quantitative genetic variation should be measured directly. PMID- 11475049 TI - Genetic variation in a host-parasite association: potential for coevolution and frequency-dependent selection. AB - Models of host-parasite coevolution assume the presence of genetic variation for host resistance and parasite infectivity, as well as genotype-specific interactions. We used the freshwater crustacean Daphnia magna and its bacterial microparasite Pasteuria ramosa to study genetic variation for host susceptibility and parasite infectivity within each of two populations. We sought to answer the following questions: Do host clones differ in their susceptibility to parasite isolates? Do parasite isolates differ in their ability to infect different host clones? Are there host clone-parasite isolate interactions? The analysis revealed considerable variation in both host resistance and parasite infectivity. There were significant host clone-parasite isolate interactions, such that there was no single host clone that was superior to all other clones in the resistance to every parasite isolate. Likewise, there was no parasite isolate that was superior to all other isolates in infectivity to every host clone. This form of host clone parasite isolate interaction indicates the potential for coevolution based on frequency-dependent selection. Infection success of original host clone-parasite isolate combinations (i.e., those combinations that were isolated together) was significantly higher than infection success of novel host clone-parasite isolate combinations (i.e., those combinations that were created in the laboratory). This finding is consistent with the idea that parasites track specific host genotypes under natural conditions. In addition, correspondence analysis revealed that some host clones, although distinguishable with neutral genetic markers, were susceptible to the same set of parasite isolates and thus probably shared resistance genes. PMID- 11475050 TI - Temporal patterns of genetic variation for resistance and infectivity in a Daphnia-microparasite system. AB - Theoretical studies have indicated that the population genetics of host-parasite interactions may be highly dynamic. with parasites perpetually adapting to common host genotypes and hosts evolving resistance to common parasite genotypes. The present study examined temporal variation in resistance of hosts and infectivity of parasites within three populations of Daphnia magna infected with the sterilizing bacterium Pasteuria ramosa. Parasite isolates and host clones were collected in each of two years (1997, 1998) from one population; in two other populations, hosts were collected from both years, but parasites from only the first year. We then performed infection experiments (separately for each population) that exposed hosts to parasites from the same year or made combinations involving hosts and parasites from different years. In two populations, patterns were consistent with the evolution of host resistance: either infectivity or the speed with which parasites sterilized hosts declined from 1997 to 1998. In another population, infectivity, virulence, and parasite spore production did not vary among host-year or parasite-year. For this population, we also detected strong within-population genetic variation for resistance. Thus, in this case, genetic variability for fitness-related traits apparently did not translate into evolutionary change. We discuss a number of reasons why genetic change may not occur as expected in parasite-host systems, including negative correlations between resistance and other traits, gene flow, or that the dynamic process itself may obscure the detection of gene frequency changes. PMID- 11475048 TI - Pollen- versus seed-mediated gene flow in a scattered forest tree species. AB - We examined the spatial distribution of maternally inherited chloroplast DNA markers over the French part of the range of Sorbus torminalis, a scattered temperate forest tree native to most of Europe. The survey by restriction analysis of polymerase-chain-reaction amplified fragments for 880 individuals distributed among 55 populations allowed the detection of 25 haplotypes. The coefficient of differentiation among populations computed on the basis of haplotype frequency (G(STc) = 0.34) was one of the lowest found in forest trees so far, and the mean within-population diversity was relatively high, indicating multiple-mother foundation events. A significant but slight geographical pattern was observed, up to distances of about 100 km. This pattern of differentiation was compared to the genetic structure of the same populations revealed by biparentally inherited markers (isoenzymes), and a new method to quantify the relative importance of seed and pollen dispersal was derived, based on isolation by-distance models. Neither pollen- nor seed-mediated gene flow was predominant in S. torminalis, a finding that differs from those for the majority of tree species studied so far. This result was most likely due to an extinction recolonization dynamics based on efficient seed dispersal strategies. The joint screening of 31 individuals of the related Sorbus aria and of 163 hybrid individuals shows that hybridization occurs predominantly in one direction and is rarely followed by cytoplasmic introgression. As a consequence, interspecific gene flow should not significantly affect the diversity dynamics within S. torminalis. PMID- 11475051 TI - Hybrid zone dynamics and species replacement between Orconectes crayfishes in a northern Wisconsin lake. AB - Hybrid zones that result in the genetic assimilation (replacement) of one species by another are underrepresented in the animal literature, most likely due to their transient nature. One such zone involves the rusty crayfish, Orconectes rusticus, and its congener O. propinquus. Orconectes rusticus was recently introduced into northern Wisconsin and Michigan lakes and streams, where it is hybridizing with and displacing resident O. propinquus. Here we report on a study investigating the dynamics of a hybrid zone between the two crayfish in Trout Lake, Wisconsin, where both the time (circa 1979) and location of the initial introduction are known. Our prediction was that hybridization should hasten the demise of O. propinquus because we expected that male O. rusticus (which are larger than congeners) would outcompete male O. propinquus for mates of both species. If hybrid progeny are unfit, then the result would be decreased reproductive output of O. propinquus females. However, we found a pattern of cytonuclear disequilibrium between allozymes and mtDNA suggesting that a majority (94.5%) of F1 hybrids resulted from matings between O. rusticus females and O. propinquus males. Also contrary to expectations, fecundity (O. rusticus and O. propinquus) and early hybrid survivorship did not differ significantly from nonhybrids. Moreover, adults of mixed ancestry were superior to both O. rusticus and O. propinquus in competition for a limiting food resource. Using a single locus model, we estimated that hybridization increases the advance of O. rusticus genes in Trout Lake between 4.8% and 36.3% above that due to the previously documented ecological interactions. Consequently, whereas hybridization may be hastening the elimination of genetically pure O. propinquus, introgression is nevertheless slowing the loss of O. propinquus nuclear genes. Although our results suggest that O. rusticus and O. propinquus may not be true species under the biological concept, their ecological differences are of great conservation importance. PMID- 11475054 TI - The effects of the minimum threshold condition for breeding on offspring sex ratio adjustment in the lesser kestrel. AB - We propose a model for sex-ratio adjustment complementary to that of Trivers and Willard. In addition to the three basic assumptions of the Trivers-Willard model, our model assumes that the sex with more variable reproductive success (normally male) is also the sex less constrained for reproduction. This assumption seems realistic, because several studies have demonstrated that poor-condition males may adopt alternative mating strategies and sire some offspring, whereas females have physiological constraints for gestation or egg production that cannot be avoided. Thus, under these circumstances, sons of both poor and good condition would be more valuable for parents than daughters, whereas daughters would be relatively more valuable than sons at intermediate condition. This model predicts, therefore, a U-shaped relationship between parental condition and offspring sex ratio. We present a case study for the monogamous lesser kestrel (Falco naumanni) that fulfills the assumptions and predictions of the model. The minimum body condition for breeding, measured as pectoral thickness, was lower for sons than for daughters. Below this minimum, males had a higher chance of breeding than females. Above this minimum, however, the lifetime reproductive success was condition dependent in males but not in females. Thus, males in better body condition attain, on average, higher reproductive success than females. Offspring sex ratio varied with the size of the father's ornaments and mother condition according to the U-shaped pattern predicted by the model. PMID- 11475053 TI - Heterozygosity-fitness correlations in rainbow trout: effects of allozyme loci or associative overdominance? AB - Previous studies with rainbow trout (Oncorhynchus mykiss) have shown that increased heterozygosity at allozyme loci is correlated with several phenotypic traits associated with fitness. We expected to find a similar effect of heterozygosity at other nuclear loci if these associations are due to loci in linkage disequilibrium with the allozyme loci (i.e., associative overdominance), rather than the allozymes themselves. We examined the association between multiple locus heterozygosity and condition factor at 10 allozyme and 10 microsatellite loci. Individuals that were more heterozygous at allozyme loci had significantly greater condition factor in two hatchery cohorts of rainbow trout (1996 P = 0.006; 1997 P < 0.001). In contrast, there was no evidence at microsatellite loci that increased heterozygosity was associated with greater condition factor. Our results suggest that the observed relationship between heterozygosity at allozyme loci and condition factor in rainbow trout appears to be due to the allozyme loci themselves, rather than associative overdominance. We cannot, however, rule out that differences in the mutation process between allozymes and microsatellites may be responsible for these observations. Regardless of the underlying mechanism, these results support the view that allozymes and microsatellites are differentially affected by natural selection. PMID- 11475052 TI - Phylogeography of the tidewater goby, Eucyclogobius newberryi (Teleostei, Gobiidae), in coastal California. AB - The tidewater goby, Eucyclogobius newberryi, inhabits discrete, seasonally closed estuaries and lagoons along approximately 1500 km of California coastline. This species is euryhaline but has no explicit marine stage, yet population extirpation and recolonization data suggest tidewater gobies disperse intermittently via the sea. Analyses of mitochondrial control region and cytochrome b sequences demonstrate a deep evolutionary bifurcation in the vicinity of Los Angeles that separates southern California populations from all more northerly populations. Shallower phylogeographic breaks, in the vicinities of Seacliff, Point Buchon, Big Sur, and Point Arena segregate the northerly populations into five groups in three geographic clusters: the Point Conception and Ventura groups between Los Angeles and Point Buchon, a lone Estero Bay group from central California, and San Francisco and Cape Mendocino groups from northern California. The phylogenetic relationships between and patterns of molecular diversity within the six groups are consistent with repeated, and sometimes rapid, northward and southward range expansions out of central California caused by Quaternary climate change. Plio-Pleistocene tectonism, Quaternary coastal geography and hydrography, and historical human activities probably also influenced the modern geographic and genetic structure of E. newberryi. The phylogeography of E. newberryi is concordant with phylogeographic patterns in several other coastal California taxa, suggesting common extrinsic factors have had similar effects on different species. However, there is no evidence of a phylogeographic break coincident with a biogeographic boundary at Point Conception. PMID- 11475055 TI - An endemic radiation of Malagasy songbirds is revealed by mitochondrial DNA sequence data. AB - The bird fauna of Madagascar includes a high proportion of endemic species, particularly among passerine birds (Aves: Passeriformes). The endemic genera of Malagasy songbirds are not allied obviously with any African or Asiatic taxa, and their affinities have been debated since the birds first were described. We used mitochondrial sequence data to estimate the relationships of 13 species of endemic Malagasy songbirds, 17 additional songbird species, and one species of suboscine passerine. In our optimal trees, nine of the 13 Malagasy species form a clade. although these birds currently are classified in three different families. In all optimal trees, the sister to this endemic clade is a group of Old World warblers including both African and Malagasy birds. The endemic Malagasy songbird clade rivals other island radiations, including the vangas of Madagascar and the finches of the Galapagos, in ecological diversity. PMID- 11475056 TI - Maladaptive mate choice maintained by heterozygote advantage. AB - Common buzzards (Buteo buteo) show a plumage polymorphism that appears to be maintained by heterozygote advantage and allows a maladaptive form of mate choice to persist. The light and dark morphs have a much lower fitness than the presumed heterozygous intermediate morph, but are replenished through Mendelian segregation in intermediate-intermediate pairs. Light and dark morphs could maximize their fitness by mating light with dark to produce all intermediate offspring, but instead choose partners of their own color, thereby producing broods of minimally fit homozygotes. Such maladaptive behavior argues forcefully against mate choice based on "good genes," and its persistence is best explained by heterozygote advantage maintaining the polymorphism coupled with nongenetic mate choice based on sexual imprinting. Modeling different patterns of mate choice shows that random mating and preference for own morph fit our data poorly, whereas preference for mother's morph yields a good fit. PMID- 11475057 TI - Genetic consequences of polygyny and social structure in an Indian fruit bat, Cynopterus sphinx. I. Inbreeding, outbreeding, and population subdivision. AB - Population subdivision into behaviorally cohesive kin groups influences rates of inbreeding and genetic drift and has important implications for the evolution of social behavior. Here we report the results of a study designed to test the hypothesis that harem social structure promotes inbreeding and genetic subdivision in a population with overlapping generations. Genetic consequences of harem social structure were investigated in a natural population of a highly polygynous fruit bat, Cynopterus sphinx (Chiroptera: Pteropodidae), in western India. The partitioning of genetic variance within and among breeding groups was assessed using 10-locus microsatellite genotypes for 431 individually marked bats. Genetic analysis of the C. sphinx study population was integrated with field data on demography and social structure to determine the specific ways in which mating, dispersal, and new social group formation influenced population genetic structure. Microsatellite data revealed striking contrasts in genetic structure between consecutive offspring cohorts and between generations. Relative to the 1998 (dry-season) offspring cohort, the 1997 (wet-season) cohort was characterized by a more extensive degree of within-group heterozygote excess (F(IS) = -0.164 vs. -0.050), a greater degree of among-group subdivision (F(ST) = 0.123 vs. 0.008), and higher average within-group relatedness (r = 0.251 vs. 0.017). Differences in genetic structure between the two offspring cohorts were attributable to seasonal differences in the number and proportional representation of male parents. Relative to adult age-classes, offspring cohorts were characterized by more extensive departures from allelic and genotypic equilibria and a greater degree of genetic subdivision. Generational differences in F-statistics indicated that genetic structuring of offspring cohorts was randomized by natal dispersal prior to recruitment into the breeding population. Low relatedness among harem females (r = 0.002-0.005) was primarily attributable to high rates of natal dispersal and low rates of juvenile survivorship. Kin selection is therefore an unlikely explanation for the formation and maintenance of behaviorally cohesive breeding groups in this highly social mammal. PMID- 11475058 TI - Genetic consequences of polygyny and social structure in an Indian fruit bat, Cynopterus sphinx. II. Variance in male mating success and effective population size. AB - Variance in reproductive success is a primary determinant of genetically effective population size (Ne), and thus has important implications for the role of genetic drift in the evolutionary dynamics of animal taxa characterized by polygynous mating systems. Here we report the results of a study designed to test the hypothesis that polygynous mating results in significantly reduced Ne in an age-structured population. This hypothesis was tested in a natural population of a harem-forming fruit bat, Cynopterus sphinx (Chiroptera: Pteropodidae), in western India. The influence of the mating system on the ratio of variance Ne to adult census number (N) was assessed using a mathematical model designed for age structured populations that incorporated demographic and genetic data. Male mating success was assessed by means of direct and indirect paternity analysis using 10-locus microsatellite genotypes of adults and progeny from two consecutive breeding periods (n = 431 individually marked bats). Combined results from both analyses were used to infer the effective number of male parents in each breeding period. The relative proportion of successfully reproducing males and the size distribution of paternal sibships comprising each offspring cohort revealed an extremely high within-season variance in male mating success (up to 9.2 times higher than Poisson expectation). The resultant estimate of Ne/N for the C. sphinx study population was 0.42. As a result of polygynous mating, the predicted rate of drift (1/2Ne per generation) was 17.6% higher than expected from a Poisson distribution of male mating success. However, the estimated Ne/N was well within the 0.25-0.75 range expected for age-structured populations under normal demographic conditions. The life-history schedule of C. sphinx is characterized by a disproportionately short sexual maturation period scaled to adult life span. Consequently, the influence of polygynous mating on Ne/N is mitigated by the extensive overlap of generations. In C. sphinx, turnover of breeding males between seasons ensures a broader sampling of the adult male gamete pool than expected from the variance in mating success within a single breeding period. PMID- 11475059 TI - Geographic patterns of genetic differentiation within the restricted range of the endangered Stephens' kangaroo rat Dipodomys stephensi. AB - Using mtDNA variation in the kangaroo rat Dipodomys stephensi, we found no support for the hypothesis that a species with an historically restricted range will exhibit low levels of genetic polymorphism and little genetic structure. Dipodomys stephensi has long been restricted to a few interior coastal valleys in southern California encompassing an area of approximately 70 x 40 km; however, we found high levels of genetic variation over much of its range and significant genetic structure both within and between regions. We also found evidence for a recent range expansion. Dipodomys stephensi is a federally endangered species that is separated from D. panamintinus, its presumed sister taxon, by a mountain range to the north. We assessed genetic variation by sequencing 645 bases of the mitochondrial d-loop from 61 individuals sampled from 16 locations across the species range and rooted their relationship using two D. panamintinus individuals. Despite its limited geographic range, the level of mtDNA variation in D. stephensi is comparable to that of other rodents, including that of the more widely distributed D. panamintinus. This variation revealed significant regional differentiation. The northern, central, and southern regions of the range differ in both the level and the distribution of genetic variation. Phylogenetic analysis revealed that the center of the range contains the most diversity of lineages, including the most basal. In this region and in the north, most haplotypes were found at only a single location (25/29), or at a pair of nearby locations (3/29). In addition, related haplotypes clustered geographically. These results are consistent with long-term demographic stability characterized by limited dispersal and high local effective population size. Further support for this conclusion is the finding of unique diversity in two northern peripheral populations, Norco and Potrero Creek (PC). However, in sharp contrast, one haplotype (CC) was found at five of 11 central and northern locations and comprised 18% of individuals sampled. The atypical distribution of the CC haplotype reflected a pattern seen more strongly in the southern region. Here the CC haplotype comprised 69% of the sample and was found at all five sampling locations. Consequently, the southern region had very low genetic variability. We propose that this dominance of CC was probably due to a local population bottleneck that occurred during a recent range expansion into the southern region. PMID- 11475060 TI - Does total reproductive effort evolve independently of offspring size? AB - In all species, patterns of reproductive allocation have important fitness consequences and therefore important implications for life-history evolution. Nearly universally, theory in this field has modeled as independent the evolution of total allocation to offspring and the subsequent division of this allocation into many small versus few large offspring. Yet, some theory and a very small amount of experimental evidence suggest that these life-history traits may be evolutionarily linked. Using comparative analyses of copepod life histories, we illustrate that rather than being evolutionarily independent these traits can be linked, in this case, across a very large clade of invertebrates. Our results indicate that a more complete understanding of the evolution of these traits will require greater consideration of simultaneous allocation decisions, rather than sequential ones, and other genetic and selective mechanisms. PMID- 11475061 TI - Estimating genetic correlations in natural populations in the absence of pedigree information: accuracy and precision of the Lynch method. AB - Usually, genetic correlations are estimated from breeding designs in the laboratory or greenhouse. However, estimates of the genetic correlation for natural populations are lacking, mostly because pedigrees of wild individuals are rarely known. Recently Lynch (1999) proposed a formula to estimate the genetic correlation in the absence of data on pedigree. This method has been shown to be particularly accurate provided a large sample size and a minimum (20%) proportion of relatives. Lynch (1999) proposed the use of the bootstrap to estimate standard errors associated with genetic correlations, but did not test the reliability of such a method. We tested the bootstrap and showed the jackknife can provide valid estimates of the genetic correlation calculated with the Lynch formula. The occurrence of undefined estimates, combined with the high number of replicates involved in the bootstrap, means there is a high probability of obtaining a biased upward, incomplete bootstrap, even when there is a high fraction of related pairs in a sample. It is easier to obtain complete jackknife estimates for which all the pseudovalues have been defined. We therefore recommend the use of the jackknife to estimate the genetic correlation with the Lynch formula. Provided data can be collected for more than two individuals at each location, we propose a group sampling method that produces low standard errors associated with the jackknife, even when there is a low fraction of relatives in a sample. PMID- 11475062 TI - Evaluation of d2, a microsatellite measure of inbreeding and outbreeding, in wolves with a known pedigree. AB - We have evaluated a microsatellite measure proposed as an indicator of inbreeding and outbreeding using a captive wolf population with known inbreeding levels and founder sources. The measure, which is based on the difference in the repeat number for microsatellite alleles within an individual, was not more predictive of the known inbreeding coefficient than microsatellite heterozygosity (it was actually less predictive). We also found no support that the measure was predictive of the level of outbreeding. However, we could not determine if the measure was predictive of very low levels of inbreeding due to matings between remote relatives. Overall, it appears that the usefulness of this measure to identify individuals on the inbred-outbred continuum beyond that of heterozygosity and identify biologically important associations with fitness related traits may be limited. We suggest that the measure be examined theoretically to determine when (and how much) the predictive value of the measure is different from that of heterozygosity for inbreeding or outbreeding levels in a variety of different scenarios. PMID- 11475063 TI - Polytomies, the power of phylogenetic inference, and the stochastic nature of molecular evolution: a comment on Walsh et al. (1999). PMID- 11475064 TI - Overview of breast imaging. AB - In summary, it is an exciting time in breast imaging with many tools being brought to bear on an ever more common problem. The challenge for this decade will be to develop optimal cost-effective strategies to use all the tools now available with minimal discomfort and disfigurement to the patient. PMID- 11475065 TI - Mammographic screening: international perspective. AB - The twentieth century saw the introduction of mammography as a diagnostic tool and its refinement as a screening method. It appears guaranteed that women who are well informed will seek mammography screening with high expectations of technical quality and accurate interpretation. More refined knowledge of breast anatomy and pathology will assist radiologists to interpret with high specificity. We will learn how to recognized more accurately normal structures and doubtful findings. We will gain experience in interpretation through faithful review of interval cancers and subtle screen-detected cancers, and will use educational tools that have the potential to improve the efficiency of education by directing attention to specific deficiencies. Mammographic screening has been advanced through the efforts of dedicated teams of physicians, scientists, and other professionals throughout the world. The international communication of ideas and discoveries will continue to challenge the boundaries of what can be accomplished in early detection as well as noninvasive therapy, and this body of knowledge will continue to be enriched by these diverse contributions. PMID- 11475066 TI - Perspective on digital mammography. AB - Digital mammography, particularly through its advanced applications, holds great promise for improved diagnostic accuracy, but the display of the images is not ideal at present. Clinical softcopy workstations are somewhat unwieldy to use, and image processing has not yet been optimized for each machine or for each clinical task. In addition, the cost-effectiveness and accuracy of the technology warrant careful study before digital mammography becomes widely disseminated and potentially replaces screen-film mammography, a technology that has been well documented to reduce breast cancer mortality. PMID- 11475067 TI - Asymmetric densities of the breast: strategies for imaging evaluation. PMID- 11475068 TI - Toward a standardized breast ultrasound lexicon, BI-RADS: ultrasound. AB - The approach outlined above for describing and reporting sonographic features of breast masses represents only the initial step in the development of a comprehensive system to enhance the accurate identification, reporting, and analysis of sonographic abnormalities of the breast. Future revisions, with validation of interobserver consistency in application of these descriptors across multiple centers, with feedback from potential users in the breast imaging community, will undoubtedly expand the utility of this effort. PMID- 11475069 TI - Review of breast MRI: indications and limitations. PMID- 11475070 TI - Illustrated breast MR lexicon. AB - A group of international breast MRI experts is currently working on a definitive lexicon for breast MRI that will incorporate both morphologic and kinetic features of lesions identified on breast MRI. The work to develop the lexicon is supported currently by the American College of Radiology (ACR). This article is aimed at introducing this material and should not be used as a definitive guide as the breast MRI lexicon is a work in progress. It is hoped that radiologists will use the terms and concepts presented here as a template to which future lexicon terminology can be added. PMID- 11475071 TI - Current status of PET in breast cancer imaging, staging, and therapy. AB - The exact roles of PET in the imaging management of patients with known or suspected breast cancer are still in evolution. For assessing primary lesions, it is sometimes possible with PET to detect cancers occult on standard methods. This could be useful in high-risk patient populations, but in dense breasts, background FDG uptake is often higher than in women with fatty breasts, making identification of lesions < 1 cm in size improbable with current technologies. Distinguishing malignant from benign primary breast disease would seem better addressed by biopsy. With a positive predictive value of FDG PET for cancer over 96%, any FDG-avid breast lesion is highly suspicious and merits biopsy. Although PET in theory should be useful for depicting multifocal disease before surgery, the limitations in detecting small lesions in the breast limit the contribution of PET at present. It is most likely that PET will have a greater role in depicting primary breast lesions as dedicated PET imaging devices for the breast evolve. For axillary and internal mammary nodal staging, results with FDG PET are variable. Small nodal metastases < or = 5 mm will be missed by PET, whereas larger ones are more likely to be detected. PET can depict internal mammary nodes, but the accuracy of the method in this setting is not known, nor is there consensus on how identifying internal mammary node metastases will change treatment. Based on the available data, for pT1 breast lesions, PET, if negative, is not an adequate replacement for sentinel node surgery or axillary dissection. Results from the multicenter trial will be of great interest. Clearly PET can stage metastatic disease well. Bone scans with 18F- are exquisitely sensitive for metastases, and FDG is also very good. However, FDG PET can miss some blastic metastases to bone so at present FDG is not capable of excluding the presence of bone metastases. PET seems very well suited to detecting recurrences in soft tissues and the brachial plexus region in particular. The utility of PET in planning the treatment of individual patients appears promising. Although results must be confirmed in larger studies, it appears safe to conclude that failure of a chemotherapy regimen to decrease FDG uptake promptly in a breast cancer portends poor response. This does not hold true for hormonal therapy. At present, labeled estrogens are not widely available and cannot be recommended for clinical use. Thus, PET has shown considerable promise in breast cancer imaging, but in the author's experience is best applied to solve difficult imaging questions in specific patients and is not recommended for routine evaluation of the breast cancer patient. However, in larger primary tumors, the ability to use PET for staging and to plan treatment response suggest it will be more widely used. Additional studies with newer PET imaging devices and FDG and other tracers will help us better determine the role of PET in routine clinical care of the patient with known or suspected breast cancer. Certainly, this represent a fertile area for translational research studies over the next several years with the potential to significantly alter the way breast cancer is imaged and managed. PMID- 11475072 TI - Sentinel lymph nodes: optimizing success. PMID- 11475073 TI - Controversies in core-needle breast biopsy. PMID- 11475074 TI - [Nuclear medicine diagnosis of patients with fever of unknown origin (FUO)]. AB - Fever of unknown origin (FUO) in immunocompetent and non neutropenic patients is defined as recurrent fever of 38.3 degrees C or greater, lasting 2-3 weeks or longer, and undiagnosed after 1 week of appropriate evaluation. The underlying diseases of FUO are numerous and infection accounts for only 20-40% of them. The majority of FUO-patients have autoimmunity and collagen vascular disease and neoplasm, which are responsible for about 50-60% of all cases. In this respect FOU in its classical definition is clearly separated from postoperative and neutropenic fever where inflammation and infection are more common. Although methods that use in-vitro or in-vivo labeled white blood cells (WBCs) have a high diagnostic accuracy in the detection and exclusion of granulocytic pathology, they are only of limited value in FUO-patients in establishing the final diagnosis due to the low prevalence of purulent processes in this collective. WBCs are more suited in evaluation of the focus in occult sepsis. Ga-67 citrate is the only commercially available gamma emitter which images acute, chronic, granulomatous and autoimmune inflammation and also various malignant diseases. Therefore Ga-67 citrate is currently considered to be the tracer of choice in the diagnostic work-up of FUO. The number of Ga-67-scans contributing to the final diagnosis was found to be higher outside Germany than it has been reported for labeled WBCs. F-18-2'-deoxy-2-fluoro-D-glucose (FDG) has been used extensively for tumor imaging with PET. Inflammatory processes accumulate the tracer by similar mechanisms. First results of FDG imaging demonstrated, that FDG may be superior to other nuclear medicine imaging modalities which may be explained by the preferable tracer kinetics of the small F-18-FDG molecule and by a better spatial resolution of coincidence imaging in comparison to a conventional gamma camera. PMID- 11475075 TI - Nonspecific bowel activity in imaging inflammation with Tc-99m labelled monoclonal anti-NCA-90 Fab' fragment MN3. AB - AIM: Since the Tc-99m labelled monoclonal anti-NCA 90 granulocyte antibody Fab' fragment MN3 (MN3 Fab') might be of interest for imaging abdominal inflammation which could be hampered by nonspecific bowel activity, we prospectively investigated the appearance of bowel activity in MN3 Fab' imaging. METHODS: Eighty consecutive patients (age range 12-85 years) referred for suspected nonabdominal, mostly musculoskeletal infection, were included. Abdominal inflammation was excluded clinically and there were no signs of inflammatory bowel disease in the patients' histories. One, 5, and 24 hours after intravenous injection of up to 1.1 GBq of MN3 Fab' planar images of the abdomen were performed. Bowel activity was graded visually using a 5-point scale. RESULTS: The one (N = 80), 5 (N = 79), and 24 (N = 52) hour images revealed 46 (10%), 162 (34%), and 173 (55%) accumulating bowel segments, respectively, in 37 (46%), 69 (87%), and 52 (100%) patients. The mean intensity score per accumulating segment was 1.1, 1.8 and 2.7 (p = 0), respectively. Relative frequencies of appearance of the small intestine were 38%, 57%, and 21%, ileocaecal region 6%, 53%, and 48%, ascending colon 5%, 67%, and 89%, transverse colon 1%, 9%, and 69%, descending colon 8%, 15%, and 67%, and rectosigmoid 0%, 4%, and 38%, respectively. Follow-up investigations in 13 patients revealed diverging uptake patterns. CONCLUSION: Nonspecific bowel activity is often present in the early and almost always and more intense, in the delayed images. Early imaging at one hour after administration seems feasible, but a loss in sensitivity has to be considered. Thus, nonspecific bowel activity can be anticipated to be a pitfall in imaging abdominal inflammation with MN3 Fab'. PMID- 11475076 TI - [Antigranulocyte scintigraphy of septic loosening of hip endoprosthesis: effect of different methods of analysis]. AB - AIM: Diagnosis of infection with help of antigranulocyte scintigraphy near body stem is difficult because of contemporary visualisation of bone marrow. Therefore, we investigated, whether it is possible to improve the accuracy in diagnosing septic loosening of hip endoprosthesis by changing the analyzing methods. METHODS: In 28 patients, the results of a visual interpretation of late scan, a visual interpretation and a quantitative interpretation of time-activity course were compared. These results were verified by histology respectively microbiology. RESULTS: Histological and microbiological verification found 14 septic loosening and 14 aseptic loosening of the hip prothesis. Therefore, sensitivity, specificity, negative and positive predictive value for the visual interpretation of late scan were 0.86, 0.57, 0.80 and 0.67. For visual and quantitative interpretation of time-activity-course we found 0.86, 0.79, 0.85 and 0.80 respectively 1, 0.93, 1, 0.93. For interobserver agreement we found kappa coefficients of 0.28 +/- 0.2 for visual interpretation of late scan, 0.48 +/- 0.17 for visual interpretation and 1.0 +/- 0 for quantitative interpretation of time-activity-course. CONCLUSION: In all investigated values quantitative interpretation of time-activity-course was superior to the other analyzing methods. Therefore, antigranulocyte scintigraphy for septic loosening of hip endoprosthesis should be interpreted quantitatively. PMID- 11475077 TI - Results of a standardized protocol for sentinel node imaging in breast cancer with Tc-99m labeled nanocolloidal albumin. AB - AIM: Of this study was to evaluate the results of a standardized protocol for sentinel node (SN) detection in breast cancer using Tc-99m labeled nanocolloidal albumin and a combined intra- and subdermal injection technique. METHODS: One hundred and fifty-five women with proven breast cancer (disease stages Tis-T2) were included. Four injections of 10 to 15 MBq of Tc-99m nanocolloid in 0.1 ml physiologic saline were administered intra- and subdermally at the 3, 6, 9 and 12 o'clock positions in the skin overlying the tumor. Planar scintigraphic images in lateral and anterior projections were obtained once between 2.5 and 18 hours after tracer administration. Guided by a gamma probe, all radioactive lymph nodes in the axilla were resected, then complete dissection followed. RESULTS: In 151 of the 155 women (97.4%), nodal tracer uptake (range 1-7 foci, average 2.2) was scintigraphically revealed. In one of these cases, drainage was only to the internal mammary lymphatic chain. Three of the 4 women with detection failure presented with histologically proven tumor infiltration of the lymphatics and axillary involvement. In 49 of the patients with visualized axillary lymph nodes (32.7%), at least one SN was metastatic. In 21 cases, this SN was the only positive node. The remaining 101 patients with negative SN included 4 cases with axillary involvement. The sensitivity of the SN with respect to the histological status of the entire axilla was thus 92.5%, the negative predictive value was 96.0%. The overall accuracy of the method was 97.3%. There was a significant difference between the number of totally detected radioactive nodes in the groups with and without nodal metastases (3.49 vs. 2.57, p < 0.01). CONCLUSION: The described protocol represents an easy reproducible and reliable method for SN detection in breast cancer that additionally allows flexible timing of surgery. Further, we found evidence that the number of scintigraphically visualized nodes also reflects the histological status of the axilla. PMID- 11475078 TI - Accuracy of the intra-operative radioguided localization of the sentinel lymph node (SLN) 24 hours after lymphoscintigraphy in patients with malignant melanoma. AB - AIM: For optimized logistics for the sentinel lymphadenectomy (SL) it might be helpful for the clinics involved if a longer time period between the lymphoscintigraphy (LS) and surgery is possible. Therefore, we investigated if a precise localization of the sentinel lymph node is possible 24 hours after LS. METHODS: 78 patients with primary malignant melanoma (MM; n = 44) or with MM pre operated by excisional biopsy (n = 34) were investigated. In 40 cases the tumor was localized on the trunk and in 38 cases on the extremities. Mean MM thickness was 2.68 mm (range: 0.29 to 12 mm). In all patients a lymphoscintigraphy (LS) with an average of 85 MBq of Tc-99m nanocolloid was performed one day prior to surgery. Immediately after tracer application dynamic data acquisition was started at a LFOV gamma camera followed by a whole body scan. With a hand-held gamma detector (C-Trak) 2, 4, 6, 8, and 24 hours after tracer administration the SLN was identified and the counts registered. RESULTS: 94 SLNs were identified in 87 lymphatic basins from which 86 could be resected. Nine MM showed two draining channels. After 24 hours 15.5% (as an average) of the initial counts could be measured in the SLN. The uptake in the SLN in pre-operated versus patients with primary tumor was statistically not significant (p = 0.4). In 16 cases (20.5%) the SLN was tumor positive. Four of those patients developed distant metastases and two died within the first year. None of the patients with negative SLN developed distant metastases or died. CONCLUSION: The remaining activity in the SLN up to 24 hours after administration is sufficient for their intra operative localization. The method of lymphoscintigraphy and localization of the SLN by a hand-held gamma detector optimizes the intra operative identification of the SLN in patients with malignant melanoma. PMID- 11475079 TI - [Radiosynovectomy for the treatment of rheumatoid arthritis of the elbow joint]. AB - The aim of this retrospective study was to evaluate the efficacy of radiosynovectomy (RSO) in patients with rheumatoid elbow arthritis. PATIENTS AND METHODS: 40 joints of 31 patients were evaluated. At the time of therapy, patients had been suffering from elbow arthritis for 17.5 months (2-72 months). 95% of the joints (n = 38) had severe daily pain or continuous pain, 97.5% (n = 39) had moderate to severe limitation of the mobility and 10% (n = 4) had severe swelling. RSO was performed by intraarticular injection of 74 MBq colloidal rhenium-186 and 15 mg triamcinolonehexacetonide. Before and six to 26 months after therapy (median follow-up 14.7 months) severity of the patients pain, mobility and swelling (transferred to a scoring system) were determined with a standardised questionnaire. A clinical re-evaluation, along with an orthrosonographical follow-up was performed in 28 joints. RESULTS: A "good to very good" overall long-term response was achieved in 80% (n = 32) of the treated joints and a temporary response in 10% (n = 4). Only 10% (n = 4) had a non satisfactory response due to advanced articular destruction. The range of motion for flexion-extension increased from 103.8 +/- 20.0 degrees to 144.0 +/- 12.8 degrees (p < 0.001). The respective scores for articular pain, impaired mobility and swelling decreased significantly (pain from 3.15 to 0.82, impaired mobility from 3.15 to 0.82, swelling from 2.40 to 0.65; p < 0.001). No deterioration or complication occurred. The effects lasted throughout the entire follow-up time for 36 joints (90%). CONCLUSION: For patients with rheumatoid involvement of the elbow joint, radiosynovectomy results in a significant decrease of articular pain and improvement of objective parameter, i.e. joint mobility. Thus, radiosynovectomy represents a feasible and effective therapeutic option for elbow arthritis. PMID- 11475080 TI - [Order of management for nuclear medicine for sentinel lymph node (SLN) diagnosis]. PMID- 11475081 TI - [Molecular imaging: dream or reality?]. PMID- 11475082 TI - [Primary diagnosis of papillary thyroid carcinoma in the framework of staging of malignant melanoma with F-18 deoxyglucose using PET]. PMID- 11475083 TI - Residents' perceptions of water quality improvements following remediation work in the Pymme's Brook catchment, north London, UK. AB - Residents' perceptions of water quality change following remediation work in the upper Pymme's Brook catchment (north London) were elicited by questionnaire and compared with monitored changes in Escherichia coli count and BMWP (The Biological Monitoring Working Party (BMWP)) score. The wider usefulness of consumer perception surveys was then discussed. Monthly data collected between 1990 and 1996 shows that both E. coli count and BMWP score improved following flushing of the foul sewerage system in 1992, but that only E. coli count improved following the subsequent completion of large-scale remedial engineering works. Local residents were surveyed regarding their awareness of the scheme, and the causes of pollution, together with their perceptions as to the effects of the engineering works and of the resulting water quality improvements. Most respondents selected and ranked indicators in a way that suggested they had an awareness of the significance of various indicators of pollution severity. Following completion of the remediation scheme, residents perceived the watercourse to contain less rubbish and sewage fungus, and to have an improved colour and smell, which corresponds favourably to the monitored improvements. However, respondents' perceptions were found to vary when the study population was sub-divided using a range of parameters. For instance, frequent observers of the brook were most likely to correctly identify sewage as the main form of pollution. These divergent perceptions suggest that there may be considerable difficulties when perception surveys are used to quantify 'benefits' following environmental improvement programmes. Nevertheless, the survey was clearly beneficial in enhancing residents' awareness of their environment and the role of their voice in its management. PMID- 11475084 TI - A comparison of alternative contingent valuation elicitation treatments for the evaluation of complex environmental policy. AB - Attempts to evaluate the economic benefits associated with complex environmental policies, using the contingent valuation (CV) method, have been dogged by controversy. In particular, debate has centred on the influence of embedding and related effects on the validity of CV willingness-to-pay (WTP) estimates. This paper discusses these effects in the context of identifying the most appropriate WTP elicitation treatment to evaluate the willingness to pay for the various elements of a multi-dimensional environmental policy. The findings of an empirical experiment demonstrate that a top-down allocation treatment, which uses independent sub-samples for individual policy components, provides the most reliable treatment to value all aspects of a multi-dimensional policy, whilst also overcoming embedding bias problems. PMID- 11475085 TI - Assessment of potential groundwater contamination sources in a wellhead protection area. AB - Determining the human health dangers from potential contamination sources, within a wellhead protection area (WHPA), requires that a risk analysis be undertaken. In this study, a desktop geographic information system and spreadsheet software are used to implement an EPA risk screening methodology for WHPAs called 'Managing Ground Water Contamination Sources in Wellhead Protection Areas--A Priority Setting Approach'. The methodology was applied to a WHPA in Gaston County, North Carolina. Results indicate that the risk of well contamination from an interstate highway and gas station with old steel underground storage tanks were comparatively high. Medium risks included a thoroughfare and highway, while low risks were assigned to machine shops, a body shop, septic systems and a gas station with new underground storage tanks and secondary containment. A sensitivity analyses of the Priority Setting Approach indicated that risk scores were extremely sensitive to hydrogeologic variables such as hydraulic conductivity. It is recommended that risk assessors utilize a range of hydrogeologic parameters to assess overall risk from each potential contamination source. PMID- 11475086 TI - Local participation, international politics, and the environment: the World Bank and the Grenada Dove. AB - The process of locating waste disposal sites in the Eastern Caribbean country of Grenada illustrates important lessons in the implementation of new international mandates to invite stakeholder participation in projects with environmental and social impacts. This case study analyzes the participatory methods and results of the World Bank-funded project in Grenada, including an unexpected shift in the policy agenda toward habitat protection for the elusive Grenada Dove, the national bird of Grenada. We conclude that the impact of new requirements for stakeholder inclusion by funding agencies such as the World Bank and Global Environmental Facility has been palpable, but mixed. As the catalysts of more participatory methods, funding agencies still must give more careful consideration to the methods by which their participatory requirements are implemented. In particular they must develop more effective knowledge of and relationships with a broader range of stakeholders than are routinely considered by existing methods, allow for and learn from unexpected contingencies, and be flexible as to project goals and methods. PMID- 11475087 TI - Community values for environmental protection in a cane farming catchment in northern Australia: a choice modelling study. AB - Choice modelling is an emerging approach to estimating the non-use values of environmental services with multiple attributes. In this paper, results are reported of a choice modelling study conducted in the Herbert River District of North Queensland to estimate the value placed on the protection of natural vegetation in areas suitable for cane production by the local community. Resource use options that vary in the level of environmental protection and the level of agricultural production were presented as a series of choice sets and respondents were asked to choose among a set of three discrete alternatives in a given choice set. The alternatives in each choice set were described by four attributes, pertaining to the area of teatree woodlands, the area of vegetation along rivers and in wetlands, regional income from cane production, and an environmental levy. The responses were analysed together with socio-economic data using a nested logit discrete-choice model to estimate the community willingness-to-pay for the protection of natural vegetation. The results indicate that the environmental values of wetlands are comparable to returns from commercial production of sugar cane and that the values of teatree woodlands are comparable to returns from extensive grazing. It is argued that land allocation policies should recognise these values in tandem with commercial benefits of production to ensure that resources are used more efficiently. PMID- 11475088 TI - Energy effects associated with e-commerce: a case-study concerning online sales of personal computers in The Netherlands. AB - The introduction of e-commerce is changing purchase and distribution patterns dramatically. One of the observed effects is that logistics become more efficient as products are directly shipped from a manufacturer or wholesaler to an end user. Another effect is that market transparency increases, which has a downward pressure on prices of many products sold via the Internet. This article addresses the energy implications of e-commerce at the micro level. This is done by quantifying the transport related energy savings in the case of a Dutch online computer reseller and by assessing the extra energy expenditure associated with increased buying power of online buyers. It is found that energy use per article sold by the online computer reseller is lower. However, taking into account indirect effects such as increased consumer buying power, there are scenarios that lead to an overall increase in energy use. PMID- 11475089 TI - Catastrophe options: an experiment in the management of catastrophic risk in the United States. PMID- 11475090 TI - ["Medical ethics patient forum--patient education and consent]. PMID- 11475091 TI - [Physicians responsibility for informed consent in Austrian law]. AB - This article considers legal aspects of the informed consent. The focus is placed on the legal basis of the doctor's duty to inform and the extent of information, the determination of which is one of the most difficult problems in practice. Finally, the importance of the duty to inform as an instrument for the avoidance of conflict and for the stabilization of the doctor-patient relationship will be discussed. PMID- 11475092 TI - [Prerequisites and limits of informed consent]. AB - The process of getting an "informed consent" is based on judicial necessity as well as on the relationship between the patient and his doctor, which has been changing remarkably over the last years. In this alliance the idea of autonomy increases, while the paternal treatment is loosing its influence continuously. Although patients have the right to decide, they also need competence in order to make a decision. Many circumstances may be responsible for a corruption of this competence. A communication based on appreciation, empathy and authenticity is able to discover such circumstances and fight against them. This kind of communication is able to prevent communication gaps, even when severe or dangerous realities have to be verbally expressed. Despite the judicial necessity the healing power of communication should get a more central place in therapy. PMID- 11475093 TI - [Prostate cancer: prevention and patient education--wishes and demands from the viewpoint of patients]. AB - Everyone is personally responsible for his/her yearly medical checks. But who is responsible for the fact that everybody knows about them? Prostate cancer is the most common malicious illness of men in Austria. How good are the chances to detect prostate cancer by various means of examinations? With the cost of one man whose cancer has been detected too late you can do the PSA test for about 10,000 men. PMID- 11475094 TI - [Global aspects of medical ethics: conditions and possibilities]. AB - A global or universal code of medical ethics seems paradoxical in the era of pluralism and postmodernism. A different conception of globalisation will be developed in terms of a "procedural universality". According to this philosophical concept, a code of medical ethics does not oblige physicians to accept certain specific, preset, universal values and rules. It rather obliges every culture and society to start a culture-sensitive, continuous, and active discourse on specific issues, mentioned in the codex. This procedure might result in regional, intra-cultural consensus, which should be presented to an inter cultural dialogue. To exemplify this procedure, current topics of medical ethics (spiritual foundations of medicine, autonomy, definitions concerning life and death, physicians' duties, conduct within therapeutic teams) will be discussed from the point of view of western medicine. PMID- 11475095 TI - [Anthropological-ethical principles of so-called patient education]. AB - Information disclosure requires veracity, authentic communication, empathy and shared decision-making. We should understand patient information as a communicative part, which is integrated into the therapeutic process. PMID- 11475096 TI - [Patient education and informed consent in hemato-oncology--limits and problems from the physician's viewpoint]. AB - Patients' information implies a complex communication process and includes more than simply giving neutral information about cancer and therapy. The goal of shared decision-making is based on the therapeutic relationship, which is rooted in the trust between doctor and patient. At the beginning of the conversation it seems paramount to optimise the setting, consisting of sufficient time, suitable space and empathy. The information should be phase-adapted and patient-centered to arrive at an informed consent about a therapeutic perspective. It seems wrong to give a plain diagnosis without therapeutic options. If possible, the next of kin should be included in the accompaniment. PMID- 11475097 TI - [Epigastric pain]. AB - It is explained how the general practitioner (= family physician) can consult a patient with epigastric pain by making use of a specific diagnostic protocol and the casugraphy. PMID- 11475098 TI - Evaluation of handicap and socio-economic status in patients with multiple sclerosis--data from a population-based survey in the sanitary area of Calatayud, northern Spain. AB - There are a few reports about handicap and socio-economic status in patients with multiple sclerosis (MS) based on epidemiological studies. The objective of our work is to evaluate handicap in patients with multiple sclerosis in the sanitary area of Calatayud, northern Spain, as well as the socio-economic situation--in comparison with patients from other parts off the world. In this study we included 34 patients with clinically definite MS found in a long-term and prospective population-based survey. For assessing the handicap degree and socio economic status we used the Environmental Status Scale (ESS) recommended by the International Federation of MS. We compared the results with those found in 1116 patients from 7 different international series. The global mean score in ESS was 9.9 (sd 9.3, range 0-31). The mean score for the item "actual work status" was 3.3 (sd 2.3, range 0-5), for the item "financial/economic status" was 1 (sd 1.7, range 0-5), for the item "personal residence/home" was 0.8 (sd 1, range 0-4), for the item "personal assistance" was 1.2 (sd 1.7, range 0-5), for the item "transportation" was 1.5 (sd 1.7, range 0-5), for the item "community health services" was 0.7 (sd 1.3, range 0-5) and for "social activity" was 1.3 (sd 1.5, range 0-4). In the comparative assessment we found that our patients were better in all items, but in the item "actual work status" where our patients yielded higher scores than those obtained in other series. Socio-economic status measured with ESS correlates well with the degree of impairment measured with EDSS and it is more favourable than previously recognized, except for "actual work status" item. The high rate of labour incapacity in our series could be due to the considerable restrictions handicapped people have to cope with in order to find employment in rural areas. PMID- 11475099 TI - [Effect of body mass index on success rate of external and internal cardioversion]. AB - Atrial fibrillation is due to complications a frequent clinical problem. Internal cardioversion shows a high effectiveness with conversion rates from up to 85%. Aim of this study was to evaluate the influence of body weight (body-mass-index [BMI]) on the effectiveness of different cardioversion procedures. PMID- 11475101 TI - [XIV National Congress of the SPIGC. Ischia, Italy, 10-12 May 2001. Abstracts]. PMID- 11475100 TI - [Study planning II--What follows the myth of the treatment refractory patient!]. PMID- 11475102 TI - [What makes a real nephrologist?]. PMID- 11475103 TI - [Hypertension and chronic renal insufficiency]. AB - In the last ten years the incidence of patients with terminal renal insufficiency has been continuously rising. A decrease in numbers is expected through vigorous treatment of systolic and diastolic blood pressure. Any antihypertensive therapy- independent of the substance group used--decreasing blood pressure, will slow down the progression of renal insufficiency. Whenever possible, an angiotensin converting enzyme inhibitor should be part of the treatment, since these drugs have been shown to be renoprotective beyond their antihypertensive effect in certain renal diseases. This review will discuss pathogenesis, treatment and the use of renoprotective drugs in hypertensive patients with chronic renal failure. PMID- 11475104 TI - [Living kidney donation--selection criteria, preparation and follow-up]. AB - Since a dialysis patient in Austria still waits on average more than two years for a renal transplant, the question of a transplant from a living donor is very interesting. We differentiate between related and non-related living donors, who are chosen on the basis of medical criteria and emotional ties. Austria's first three kidney transplants from related donors were performed in 1967. Since then a total of 317 kidneys from genetically related donors have been transplanted until December 31, 2000. Transplants from non-related living donors were performed once in 1982, once in 1990 and since 1995 in a steadily increasing number each year, until they reached 47 by December 31, 2000. The United Network for Organ Sharing calculated the ten-year survival rate for functional grafts for a four-year period (1995-1998) in more than 30,000 renal transplant recipients from HLA identical twins, non-related living donors, parent and cadaver donors. As anticipated, this study demonstrates that HLA-identical twins (n = 1,581) have the most functional grafts (81%), followed by non-related donor-recipients (n = 1,704) at 67% despite their often poor HLA match, parent-child transplants (n = 2,428) at 62% and cadaver renal grafts (n = 26,178) at 50%. Therefore, medical aspects as well as influences from the psychosocial environment would appear to be decisive for transplantation success. Thus, when choosing from several possible living donors it is absolutely justifiable to choose a donor with a poorer HLA match but good emotional ties. Such a choice requires strict selection criteria, and surgical preparation and follow-up demand the greatest care. While the criteria given in this paper are meant to be guidelines to help in deciding for a liver donor, they certainly do not rule out a different approach following critical reflection and participation by the affected parties, namely donor and recipient, as well as their advisors, nephrologist, transplant surgeon and psychotherapist. At the same time we need to make every effort to further intensify the use of cadaver kidneys. Only in this way can we ensure optimal implementation of all the resources available to us for supplying renal grafts to dialysis patients. PMID- 11475105 TI - [Value of sonography in kidney transplantation]. AB - Colour coded duplex sonography (CCDS) is the method of choice in diagnostic monitoring of kidney transplants. Lack of radiation exposure, easy accessibility and high resolution have contributed to the fact that CCDS is used to monitor kidneys after transplantation. However, the value of CCDS in diagnosis of rejection is still a matter of debate. CCDS is used to evaluate perfusion of transplanted kidneys, detect renal artery stenosis or real vein thrombosis and arteriovenous fistulas. The value of recent technical advances, such as 3D Sonography, contrast harmonic imaging and tissue harmonic imaging, is currently being tested. This article focuses on indications and limitations of CCDS in the diagnosis of functional impairment of transplanted kidneys. PMID- 11475106 TI - [Pathogenesis and clinical course of hereditary nephropathies]. AB - Hereditary diseases have to be considered in the differential diagnosis of many kidney diseases. The kidney can be affected by systemic metabolic diseases such as primary hyperoxaluria or Fabry's disease. Inborn errors of the coagulation cascade or the complement system may cause familiar forms of the hemolytic uremic syndrome. Of central interest are hereditary cystic kidney diseases with autosomal dominant polycystic kidney disease as its most prominent example. Hereditary forms of the nephrotic syndrome are usually caused by abnormalities of podocyte function. Alport's syndrome is a classical example of a basement membrane disease. Of special interest are hereditary defects in tubular transport mechanisms such as carrier defects affecting sodium reabsorption along the tubulus. PMID- 11475107 TI - [Immunosuppressive therapy and bone metabolism after kidney transplantation]. AB - The success of transplant medicine due to improvements of immunosuppressive therapy has led to a significant increase of patient and organ survival. With the increasing number of transplantations, however, long term complications, often affecting the skeletal system, are becoming more frequent. Bone alterations often exist prior to transplantation in patients with chronic renal failure. There are two types of renal osteopathy, including "low-turnover bone disease", consisting of osteomalacia, and adynamic bone disease, and "high-turnover bone disease" due to the development of secondary hyperparathyroidism. Many patients show evidence of both disorders (mixed bone disease). During the first months after transplantation patients lose bone mass rapidly. One of the major factors responsible for the development of osteoporosis is thought to be the intensive immunosuppressive therapy during that period, steroids in particular seem to play an important role. To what extent other medications influence bone metabolism has not been established. Currently there are no studies about a standardized therapy and treatment relies mainly on experience with other forms of osteoporosis. PMID- 11475108 TI - Use of cartilage grafts in the treatment of laryngotracheal stenoses and defects in children. AB - Stenosis of the laryngotracheal section is in most cases a sequel of intensive care treatment. Most of the children presenting with such stenoses need tracheostomy that in consequence impedes their physical, social and language development. Surgical treatment of stenosis and closure of tracheostomy as early as possible should be strived for. Reconstructive techniques using autogenous cartilage grafts have proved very effective. The aim of our study was to examine the influence of different methods on the structure and function of reconstructed airways. In 23 children between 2 and 16 years suffering from laryngotracheal stenosis we reconstructed the stenotic segments by insertion of autogenous cartilage grafts. Enlargement was achieved by cartilage implantation as a stent into the split cricoid plate in 16 children; in 3 cases we performed enlargement of the anterior wall and in 5 cases instable lateral walls of the trachea had to be reinforced. One child had to be treated with all three techniques at once. In another case the overstretched and soft posterior tracheal wall had to be stabilized. Enlargement of the cricoid plate has proved excellent in cases of a high degree of stenosis of the posterior commissure (Type III and IV of the Cotton classification). Subglottic stenoses Type II can be treated with anterior insertion of cartilage. The use of cartilage in surgical treatment of laryngotracheal stenosis in childhood leads to safe and sufficiently satisfactory results that do not hamper further development of either larynx or trachea. PMID- 11475109 TI - Thyroid tumors in children and adolescents: preoperative study. AB - Several studies indicate that in young patients (less than 21 years of age at the time of diagnosis), the prognosis of thyroid carcinoma (TC) is more favorable than in older patients. However, a more radical treatment approach is recommended in children and adolescents due to the higher prevalence of local lymph-node involvement in these cases. Since the extent of primary surgical treatment is closely related to the overall prognosis, preoperative diagnosis becomes essential in the management of thyroid neoplasms in young patients. In this retrospective study (1987-1998), we analyzed a surgical series of 50 children and adolescents with thyroid nodules in an attempt to establish the role of diagnostic studies in detecting malignant lesions prior to surgery. Our diagnostic protocol for evaluating thyroid nodules was based on clinical evaluation, measurement of thyroid-hormone and thyroglobulin (TG) levels, anti-TG and anti-TPO antibody titers, calcitonin, CEA, and TPA levels, sonography, scintigraphy, and fine-needle aspiration cytology (FNAC) of the thyroid nodules and any enlarged lymph nodes. Eleven of the 15 cases of histologically confirmed carcinoma were preoperatively identified as malignant lesions with the aid of FNAC. The authors conclude that the preoperative work-up of children and adolescents with thyroid nodules requires the collaboration of an experienced team of professionals, and recommend FNAC as the initial test. PMID- 11475110 TI - Is intestinal ischemia a risk of laparoscopy? An experimental study in rabbits. AB - BACKGROUND AND PURPOSE: In the literature, there are few reports documenting intra-abdominal organ necrosis following laparoscopic procedures. This experimental study was planned to investigate whether intestinal ischemia develops during laparoscopic procedures and if laparoscopy could cause intestinal necrosis. MATERIAL AND METHODS: Two experimental groups, each consisting of 10 adult New-Zealand rabbits, were used in this study. The first group comprised the study group which had pneumoperitoneum, the second group comprised the normal animals serving as the controls. A cervical tracheostomy was performed to achieve successful general anesthesia in both groups. In the study group, intraperitoneal CO2 insufflation was carried out and intraabdominal pressure (IAP) was adjusted so as not to exceed the arterial blood pressure. After 20 min high IAP period, the intraabdominal gas was aspirated. Five minutes later, samples of both small intestine and colon tissue were taken. In the control group, tissue samples were taken 25 min after anesthesia was achieved. Xanthine oxidase (XO) and malondialdehyde (MDA) levels were measured as indicators of intestinal ischemia and lipid peroxidation in the intestinal tissues. Statistical analysis was done to compare the XO and MDA levels of the small intestines and colons of both groups. RESULTS: The mean colonic XO levels were 1.323+/-1.17 and 0.217+/-0.27 (U/mg protein) in study and control groups, respectively. This difference was statistically significant (t = 2.60, p<0.05). The other comparisons with regard to XO and MDA levels were statistically not significant. CONCLUSION: Our results demonstrate that intraperitoneal CO2 insufflation in which intraabdominal pressure was adjusted to be lower than arterial blood pressure may affect oxygenization of the colon. PMID- 11475111 TI - Urinary trypsin inhibitor and biliary atresia--providing protection for the liver? AB - Urinary Tyrosine Inhibitor (UTI) is produced in the liver and excreted in urine hepatic inflammation, infection or malignancy. We assess the possible implications of UTI in biliary atresia (BA). Liver function was used to divide 34 postoperative BA patients into 2 groups: Group 1 (n=25), anicteric (total bilirubin [T-Bil] <2.0 mg/dl); and Group 2 (n = 9), icteric (total bilirubin >2.0 mg/dl) with abnormal liver function test results, and repeated episodes of cholangitis. 26 age-matched subjects with no history of liver disease acted as controls PMID- 11475112 TI - The role of recombinant human erythropoietin in lipid peroxidation and platelet activating factor generation in a rat model of necrotizing enterocolitis. AB - In the present study we examined the effect of recombinant human erythropoietin (rhEPO) on intestinal malondialdehyde (MDA) as an index of lipid peroxidation, related to iron-catalysed free radical reaction and platelet-activating factor (PAF) synthesis in the experimental model of necrotizing enterocolitis (NEC). Three groups, each consisting of eight 1-day-old Wistar albino rat pups, were studied; Group 1, hypoxia-reoxygenation; Group 2, hypoxia-reoxygenation and rhEPO pretreatment; Group 3, control. rhEPO was given 750 U/kg/week by intraperitoneal injection three times a week for 2 weeks. On day 15th of life, hypoxia was induced by placing rat pups in a 100% CO2 chamber for 5 min. After hypoxia, the rat pups were reoxygenated for 10 min with 100% oxygen and returned to their mothers. All pups were killed at 4h following hypoxia-reoxygenation. The abdomen was opened and representative samples of injured areas were taken for histopathologic examination. MDA and PAF levels were determined in the intestine. Significantly increased intestinal MDA content was found in Group 1 rat pups compared to Group 2 and Group 3 pups (p < 0.001 and p < 0.001, respectively). However, PAF concentrations were highly elevated in the intestine of Group 1 and Group 2 pups (p>0.05) when compared to the intestine of Group 3 pups (p < 0.001 and p < 0.001, respectively). Histopathologic findings did not differ between Groups 1 and 2. The present study demonstrates that oxygen-derived free radicals and PAF are involved in the pathophysiological mechanism of the development of NEC. This study also shows that administration of rhEPO significantly decreases lipid peroxidation; however, PAF generation was not inhibited in hypoxia-induced bowel necrosis. PMID- 11475113 TI - Testicular descent: when to interfere? AB - Spontaneous descent of testes after birth can occur in up to 70% of cases, yet the factors contributing to it are still controversial. This study aims to evaluate factors contributing to spontaneous descent of palpable undescended testes. Eighty-four newborns with 126 palpable undescended testes (42 unilateral and 42 bilateral) were followed up for a period of one year to study the occurrence and time of testicular descent and its relation to gestational age, birth weight, uni- or bilaterality and levels of FSH, LH and testosterone. A total of 58 testes (46%) descended between 3 and 6 months. Spontaneous descent occurred in 10 premature patients (14 testes 63%) compared to 44 testes of full term patients (43%). Descent occurred in 14 unilateral undescended testes (33%) compared to 44 (52%) in bilateral cases. In patients with spontaneous testicular descent there was postnatal peak of LH and testosterone at 2 to 3 months of age which returned to basal level at 6 months of age. In patients with permanent undescended testes the peak of LH and testosterone was very low and almost absent in some of them; no significant difference was found between the mean values of FSH in both groups. No spontaneous testicular descent occurred after the 4th month in the full-term group, whereas in the pre-term group spontaneous descent occurred up to 6 months of age. This study concluded that spontaneous descent of palpable undescended testes is closely related to the presence of LH and testosterone surge. Therapy of undescended testes should start at 4 months of age in a full-term baby and at 6 months of age in a pre-term baby. PMID- 11475114 TI - Experience with laparoscopy in nonpalpable testis. AB - AIM: The aim of this study was to investigate the feasibility and benefits of diagnosis and interventional laparoscopy in those paediatric patients with nonpalpable testes (NPT). PATIENTS AND METHODS: Between 1992 and 1999, 75 patients with NPT (mean age 3 years, range 6 months to 14 years) were treated. 86 testes were evaluated. RESULTS: According to the laparoscopic findings 4 groups of testes were identified: Vanishing testis (n = 32), low abdominal testis (< 2 cm to the internal ring) (n = 26), high abdominal testis (> 2 cm to the internal ring) (n = 24) and intersex patients (n = 4). Of the first group, 19 testes (one bilateral) had blind-ending spermatic cord and vessels and if an atrophic testicular tissue was identified, it was removed laparoscopically. For those with spermatic cord and vessels beyond the internal ring (13 testes), atrophic testes were removed through a high scrotal incision. 19 testes of the second group had a laparoscopy-assisted orchidopexy. In the same group a laparoscopic orchidopexy was performed on 7 testes. 24 testes in the 3rd group had a Fowler-Stephens (FS) stage 1 and 18 testes had a laparotomy performed for FS stage 2 procedure (laparotomy and orchidopexy) after 6 months. At laparotomy there was no evidence of testicular atrophy in all but one testis, which was removed and the FS stage 2 procedure was completed in 17 testes. The follow-up period was between 6 months and 4 years, and two more testicular atrophies were noted after FS stage 2. The results were satisfactory in 15 out of 18 testes (83%). In the intersex group, the patient with testicular feminization underwent laparoscopic orchiectomy. The other patient with bilateral nonpalpable testis was identified as having an uterus and two intraabdominally located gonads on laparoscopy and gonadal biopsies were obtained for diagnosis. Histology demonstrated bilateral ovotestes, confirming the diagnosis of a true hermaphrodite. CONCLUSION: We are of the opinion that laparoscopy decreases the number of laparotomies in NPT, allows a single-stage procedure in low abdominal testis, and facilitates clip ligation of the testicular artery in high abdominal testis. Laparoscopy also provides diagnostic and therapeutic options for vanishing testis and intersex patients. PMID- 11475115 TI - Sacrococcygeal teratoma: results of a retrospective multicentric study in Belgium and Luxembourg. AB - Eighteen patients, operated upon for sacrococcygeal teratoma in 7 different centres in Belgium and Luxembourg between 1992 and 1996, were reviewed. From an epidemiological point of view, this series compares very well to others. Although excellent results were obtained, with all patients surviving, some imperfection in diagnosis, timing of delivery and of operation, and in operative technique was observed. Therefore, it is stated that for optimal treatment of sacrococcygeal teratoma to be achieved, these cases should be treated in just a very few centres of neonatal surgery. PMID- 11475116 TI - Anal canal duplication in infants and children--a series of 6 cases. AB - The authors present a series of six anal canal duplications (ACD), duplications of the alimentary tract located along the posterior side of the anal canal, with a perineal opening just behind the anus. Five asymptomatic duplications were diagnosed before the age of one year, by simple perineal inspection. A twelve year-old girl presented with perineal and anal pains and diarrhoea. Fistulography revealed a tubular structure in five cases and a cystic structure in one case, behind the normal anal canal, in one case communicating with it. A presacral sacrococcygeal teratoma was found in two children and in one case it was visualised by preoperative US in an infant with a lumbosacral myelomeningocele. Surgical excision was performed by a perineal approach in 5 cases, by a combined sacral and perineal approach in the last case, because of the associated teratoma. Non-invasive preoperative investigations, consisting of a pelvic X-ray, US examination, barium enema and fistulography, are sufficient in most cases; MRI is reserved for special indications. Surgical treatment restores a normal perineal aspect, without sequelae, and avoids complications like those described in other types of digestive duplications: infection, ulceration, bleeding, malignant changes during later adult life. Associated anomalies are frequently described in the literature, especially presacral tumours (16%) and anorectal malformations (21%); they can influence the management, the surgical approach and the functional prognosis. PMID- 11475117 TI - Anorectal atresia. An experimental model in the rat. AB - Aiming to assess the development of the anorectal sphincteric apparatus and whether it is influenced by the presence of an intestinal conduit through the muscular complex, we seek to develop an experimental model of anorectal atresia in foetuses of female pregnant rats, given orally the herbicide ethylenethiourea (ETU). Six 3-months-old, 300 g female Wistar rats were included in the study group (SG). The foetuses of another unexposed rat formed the control group (CG). In the SG 54, foetuses were alive, all had hypoplasia of the tail and 47 (87%) had no visible anal orifice. The 18 foetuses of the CG were normal. The histological study was performed at the level of the pubic symphysis. Type I anorectal atresia was defined when this section demonstrated an intestinal conduit and Type II when only a muscular cord was found, without any intestinal tube. This study was completed in 15 (32%) of the SG, 9 (60%) with anorectal atresia Type II and 6 (40%) with atresia Type I, and in 11 animals (61%) of the CG. The thickness of the sphincter muscle was larger in the CG (260.47+/-35.32 micro) than in the SG (208.98+/-55.01 micro), p<0.05, but this difference was due to the animals showing atresia Type II. To conclude: ETU had a teratogenic effect. The foetuses with atresia, particularly Type II, had a decreased development of the muscular complex. The presence of an intestinal conduit does not significantly influence the development of the muscular complex. PMID- 11475118 TI - Pleuropulmonary blastoma in congenital cystic adenomatoid malformation: report of a case. AB - Pulmonary blastoma is a rare malignant tumor seen in both adults and children. Approximately only 25% of cases occur in pediatric patients, many of whom affected by a congenital pulmonary cystic lesion. The clinical features, radiological findings and management of a 3-year-old boy affected by a pulmonary blastoma which arose in a congenital cystic adenomatoid malformation are reported, and an extensive review of the literature is also made. Because of the well-known tendency of cystic pulmonary diseases to develop malignancies, authors recommend the surgical excision of these kind of lesion or at least their close radiological follow-up. PMID- 11475119 TI - Multiple ipsilateral congenital diaphragmatic pathologies: rarities to consider. AB - BACKGROUND: Bilaterality of congenital diaphragmatic pathologies is well documented in the English literature. Nevertheless, ipsilateral simultaneous congenital diaphragmatic pathologies, though rare, are worth revisiting to shed more light into their embryology and management. METHODS: A review of the literature for reported cases of multiple ipsilateral congenital diaphragmatic pathologies was performed. Their findings, management and outcomes were classified and reviewed, in addition to reporting and comparison with a new case from our medical center. RESULTS: Two patients were adult females with multiple defects in the right central tendon of the diaphragm. The remaining four were all pediatric cases. Two had double congenital hernial defects on the same side and two had the defect associated with ipsilateral diaphragmatic eventration. Our case is the third one with diaphragmatic eventration, but the first being associated with a posterolateral defect on the same side. Its presentation with acute rectal bleeding proved difficult to diagnose. CONCLUSION: There are various postulated theories concerning the pathogenesis of ipsilateral pathologies during embryologic development of the diaphragm. Management of asymptomatic congenital eventrations remains debatable. However, when symptoms become apparent, ipsilateral defects though rare, should be considered and prompt surgical management through a thoracotomy or laparotomy approach is recommended. PMID- 11475120 TI - Acute perforated neonatal appendicitis associated with chorioamnionitis. AB - Perforation of the appendix is rare in term neonates and even more rare in prematures. In most of the reported cases it is a complication of necrotizing enterocolitis (NEC) but can also complicate Hirschsprung's disease (HD), cystic fibrosis (CF) or acute appendicitis (AA). We present a pre-term newborn, born to a mother who had bacteriaemia associated with chorioamnionitis. On his sixth day of life he had clinical and radiological signs of perforated abdominal viscus. Laparotomy and histologic findings indicated acute perforated appendicitis. The association of acute perforated appendicitis with chorioamnionitis points to the role of maternal foetal infection in the aetiology of neonatal acute appendicitis. PMID- 11475121 TI - Transitional cell carcinoma of the urinary bladder following exposure to cyclophosphamide in childhood. AB - With the increasing survival of children with malignancies, the long-term effects of chemotherapies are now becoming important and one of these dreadful long-term complications is the development of a second malignancy. Described here is a case of transitional cell carcinoma of the bladder developing 8 years following exposure to cyclophosphamide for the treatment of embryonal rhabdomyosarcoma. PMID- 11475122 TI - Paediatric surgeons websurfers bulletin. PMID- 11475124 TI - Biological therapies for inflammatory bowel diseases: are we finally going to modify the disease course? 26-27 January 2001, Leuven. Symposium proceedings. PMID- 11475123 TI - Detection of Helicobacter pylori in dental plaque and gastric biopsy samples of Turkish patients by PCR-RFLP. AB - Helicobacter pylori is regarded as an important pathogen playing a key role in the pathogenesis of peptic ulcer. Different studies about the mode of transmission of the microorganism report conflicting results about dental plaque as the source of H. pylori infection. In the present study we aimed to detect the presence of H. pylori in dental plaque of Turkish patients by polymerase chain reaction (PCR) and if any to do typing by restriction fragment length polymorphism (RFLP) analysis. Fifty dyspeptic patients, to whom upper gastrointestinal endoscopy was performed, were included in the study. Dental plaques were obtained before endoscopic examination. Both dental plaque and gastric biopsy samples were amplified with Ure A and Cag A gene primers. There were no positive dental plaque samples even in the 23 patients whose gastric biopsy specimens were positive. Our findings showed that there is not a correlation between dental presentation of the microorganism and H. pylori gastritis. PMID- 11475125 TI - The changing epidemiology of inflammatory bowel diseases. AB - Epidemiological studies of inflammatory bowel disease are important in order to find possible clues to the still unknown etiology, as well as for the planning of the health service. In supplement to short-term studies, reporting the actual state of the disease, long-term studies are requisited to follow up the development of inflammatory bowel disease. Inflammatory bowel disease has become more frequent during the past decades, with an approximate prevalence of 0.5% in the northern part of the world. A changed disease panorama can be noted with increasing median age at diagnosis, a growing entity of patients with colorectal Crohn's disease and a shift towards more distal ulcerative colitis. The increased age at diagnosis is attributed to a postponement of the age-specific incidence peak. While the difference between gender has levelled off, difference still exists in young adults in Crohn's disease and in elderly in ulcerative colitis. Colorectal cancer has become a matter also in Crohn's disease. PMID- 11475126 TI - Genetics of inflammatory bowel disease--an update. PMID- 11475127 TI - Biologicals: principles, techniques and mechanisms of action. AB - Biological agents for the treatment of IBD are the result of both the explosion of knowledge precipitated by the techniques of molecular biology, and by the ability to use these same techniques to produce agents. Thus, there has been a greatly facilitated translation of basic knowledge into clinical therapy. An astounding number of biologic agents are currently in development for the treatment of IBD and other immune-mediated conditions. These include native microbiologic preparations isolated for beneficial properties, recombinant cytokines and anticytokines, monoclonal antibodies, antisense oligonucleotides, and in the future, somatic gene therapy. This work seeks to describe the principles and techniques of biologic agent development, as well as prime sites of action targeted by these agents. Recent advances in the techniques of molecular biology have made possible unprecedented progress in the treatment of many conditions. The techniques of molecular biology have provided new methods of drug discovery and at the same time have elucidated new therapeutic targets. Most notable has been the progress made in the treatment of chronic inflammatory and immune mediated conditions, including inflammatory bowel disease. This paper is intended to highlight the methodological principles behind biologic agents, methods of discovery and production, and to highlight potential therapeutic targets for these new agents. PMID- 11475128 TI - Strategies targeting tumor necrosis factor in Crohn's disease. AB - Tumor necrosis factor plays an important role in mediating the inflammation of Crohn's disease. Strategies aimed at reducing tumor necrosis factor in patients with Crohn's disease include the mouse/human chimeric monoclonal antibody infliximab, the humanized monoclonal antibody CDP571, the human recombinant tumor necrosis factor receptor fusion protein etanercept, and the small molecule thalidomide. Infliximab is effective for treating active Crohn's disease, maintaining remission, and closing fistulas. Side effects occurring in patients treated with infliximab include human anti-chimeric antibodies, infusion reactions, formation of autoantibodies, and rarely drug induced lupus. CDP571 is effective for treating active Crohn's disease, steroid sparing, and possibly for closing fistulas and maintaining remission. Side effects occurring in patients treated with CDP571 include anti-idiotype antibodies, infusion reactions, and formation of autoantibodies. Pilot studies have suggested that etanercept and thalidomide may also be beneficial. Anti-tumor necrosis factor therapies are effective for the treatment for Crohn's disease. PMID- 11475129 TI - Tissue effects of anti-TNF therapies. PMID- 11475130 TI - Biological therapies of inflammatory bowel disease. PMID- 11475131 TI - Standard immunosuppression in IBD: current practice. PMID- 11475132 TI - Crohn's disease therapy: step up or top down therapy. AB - In most Crohn's disease scenarios a step-up approach has been used to initiate therapy according to disease severity. However, this approach has ignored the concept of inductive therapy followed by maintenance treatment to prevent relapse. It is apparent from clinical trials that the success of maintenance approaches will depend upon the inductive therapy. Furthermore, the step-up approach often results in steroid-dependent or refractory disease. The potential of aggressive initial therapy followed by step-down maintenance therapy affords the potential to modify the disease course, a possibility that has not been accomplished by current therapeutic algorithims. PMID- 11475133 TI - Biologics in peri-operative management of Crohn's disease. AB - The role of biologics for peri-operative Crohn's disease will remain speculative until the accumulation of clinical trial data. The potential positioning of peri operative biologic therapies includes the prevention of surgery, minimizing surgical morbidity or prevention or delaying post-operative disease recurrence. To date, there is limited, indirect data regarding the potential for infliximab to prevent surgery via the effective treatment of refractory disease and the improvement in fistulizing disease. Although there is no controlled data regarding the impact of biologics on surgical morbidity, patients with stricturing disease have undergone resection without added morbidity. Post operative trials are certainly needed to ascertain the impact on the inevitability of disease recurrence. PMID- 11475134 TI - Can we influence fibrosis in Crohn's disease? AB - Despite recent advances in the therapy of active Crohn's disease (CD) fibrostenosis remains a challenging complication of the disease. Transmural inflammation of CD is associated with phenotypic switch of the mesenchymal cells resulting in proliferation and collagen deposition. Both resident myofibroblasts and smooth muscle are candidate progenitor cells of the fibrogenic cells in CD stenoses. The principal growth factors involved in intestinal fibrosis have not been identified although TGF-beta 1 and 2, PDGF and IL-1 may be involved. Research aimed at elucidating the basic mechanisms underlying fibrosis in the gut has been hindered by the lack of an adequate animal model. Recently, however, new rodent models of chronic inflammation with distinct fibrosis have been described. Cell culture research has provided more information about possible pathways that may limit uncontrolled mesenchymal proliferation in the inflamed intestinal wall. The modulator role of neurotransmitters such as VIP, nitric oxide and prostaglandines is an important target for therapeutic intervention. Interfering with the phenotypic switch of mesenchymal cells may offer new therapeutic perspectives in the prevention of fibrostenosis. Further in vitro and animal studies as well as restenosis prevention studies are needed to develop pharmacological tools in the prevention of Crohn's disease fibrostenosis. PMID- 11475135 TI - Crohn's disease, ulcerative colitis or indeterminate colitis--how important is it to differentiate? AB - In most patients coming to the general practitioner or specialist with a history of bloody diarrhoea, bacteria or drugs are the most likely causative agents and it will be possible to make a diagnosis fairly easily. Because of differences in treatment, ulcerative colitis (UC) and Crohn's disease (CD) must however seriously be considered especially in younger patients, with severe symptoms and whenever the history is prolonged. A variety of colitides may indeed be clinically confused with UC and CD. Pathological mimics that should not be missed include infectious diseases such as Campylobacter colitis, yersiniosis, amoebiasis and others; drug-induced diseases (due to nonsteroidal antiinflammatory drugs...); diverticular disease-associated colitis; intestinal endometriosis; intestinal vasculitis and Behcet's disease and iatrogenic conditions such as graft-versus-host-disease and radiation colitis. In most situations a precise diagnosis of these conditions should be possible when all data are available. The term "indeterminate colitis" is used, when a diagnosis of chronic idiopathic inflammatory bowel disease (IBD) is suggested, but the differential diagnosis between UC and CD can not be solved. This occurs in approximately 5% of all patients with IBD. Diagnostic problems can occur in acute fulminant colitis, acute prolonged colitis, chronic relapsing disease and pouchitis. Indeterminate colitis is essentially a temporary diagnosis. Surgical and medical treatment of these patients can be difficult. When surgical treatment is indicated, the type of surgery must be seriously considered. The clinical course of patients with indeterminate colitis is usually more severe when compared with classical UC and these patients require often more severe medical treatment. Diagnostic problems can also arise in longstanding IBD, either UC and CD. Relapse of symptoms can be due to intercurrent infection (CMV is one of the candidates). Medical treatment can influence the microscopic features and induce a discontinuous inflammation in UC, reminiscent of CD. In cases of doubt, the original biopsies should be reviewed to ascertain the diagnosis, and orient treatment. PMID- 11475136 TI - Cyclosporine in ulcerative colitis: state of the art. AB - Forty percent of patients with severe ulcerative colitis will fail to respond to intravenous corticosteroids. Cyclosporine and other calcineurin inhibitors offer an alternative to colectomy for these patients. Intravenous cyclosporine will induce remission within 14 days in 50-80% of patients who fail intravenous corticosteroids. The long-term response rates for responding patients are 40-60%. Subsequent maintenance therapy with azathioprine or 6-mercaptopurine is recommended at the present time, although the uncontrolled studies underlying this observation have yielded variable results. Toxicity occurs frequently in patients treated with high dose cyclosporine, and there is a small risk of opportunistic infection and death. Pilot studies have suggested that the microemulsion cyclosporine formulation Neoral and tacrolimus may also be of benefit in this patient population. Additional studies to determine the dose response of intravenous cyclosporine, to determine the role of azathioprine for maintenance, and to determine the efficacy of Neoral and tacrolimus are needed. PMID- 11475137 TI - Biological therapies for ulcerative colitis. AB - Biological therapies are being increasingly investigated for the treatment of inflammatory bowel disease. However, a great deal more study has been devoted to studies of Crohn's disease rather than ulcerative colitis. Ulcerative colitis, like Crohn's disease, represents an area of high clinical need, particularly for those patients who have disease inadequately responsive to corticosteroids and 5 aminosalicylates. The distinct anatomic distribution of inflammation in ulcerative colitis represents an important model for study, with the entire involved mucosa entirely accessible to endoscopy. In addition, there is an opportunity for local delivery of biologic agents in left-sided disease. Distinct pathogenetic factors in ulcerative colitis raise the possibility of therapies quite different from those used in Crohn's disease. This work describes the current state of knowledge regarding biological therapy in ulcerative colitis. The role of probiotic therapy, and studies of cytokine-directed therapies, therapies targeting adhesion and recruitment, and restitution and repair are described. PMID- 11475138 TI - Safety of biologics: current issues and future concerns. PMID- 11475139 TI - Restorative resections for ulcerative colitis. PMID- 11475140 TI - Refractory pouchitis. PMID- 11475141 TI - Advancement flap plasty for the closure of anal and recto-vaginal fistulas in Crohn's disease. AB - The management of anal fistulas in patients with IBD continues to be extremely challenging and, indeed, somewhat frustrating. Despite a global closure rate of about 75%, all patients should be informed about the risk of infection, early failure, eventual temporary disfunctioning stoma and the possibility of late recurrence (about 15%). Closure of a RVF in Crohn's disease should not be considered an easy undertaking, especially in patients with several Crohn localisations. The technique can be adapted to the local situation. Construction of a temporary stoma is not mandatory. However, stoma construction seems to be beneficial when extensive perianal or recto-vaginal dissection including eventual tissue interposition is required. Advancement flaps are an attractive surgical alternative for the management of all anal transsphincteric fistulas, also in Crohn's disease, because sphincter architecture and function are well preserved. Improved medical treatment and the changed approach from conservative to reparative surgery may well have resulted in a decreased need or at least in a delay of the need for a proctectomy. Although the surgical principles of advancement flap techniques are sound, these techniques have not been used for many decades. Skills needed, problematic approach, suboptimal quality of local tissues have contributed to its selective use and to the absence of prospective randomised studies. PMID- 11475142 TI - Autoimmune pancreatitis mimicking cancer of the head of pancreas: report of two cases. AB - Autoimmune pancreatitis has been characterised in 1995, but only a few cases have been published since then, most of them from Japan. This report describes the cases of two Belgian male patients who presented with isolated obstructive jaundice. Radiological imaging studies were highly suggestive of carcinoma of the head of pancreas and both patients underwent uneventful cephalic pancreaticoduodenectomy with portal vein resection. Pathological analysis of the removed tissues suggested an autoimmune process in both cases. Both patients had hyper-gammaglobulinemia and antinuclear antibodies, but failed to show evidence of any other autoimmune disease or cause of chronic pancreatitis. In both cases final diagnosis was autoimmune pancreatitis. Preoperative clinical suspicion of this diagnosis is mandatory and may avoid unnecessary surgery in future cases. PMID- 11475143 TI - Hepatic myelopathy after splenorenal shunting: report of one case and review of the literature. AB - Hepatic myelopathy is a rare complication of cirrhosis, usually associated with surgical or spontaneous porto-systemic shunts. Its pathophysiology is unknown. It is characterized by a motor involvement of the lower limbs without clinical sensory abnormality, leading to spastic paraparesis. These neurological features are related to a symmetric loss of myelin in the lateral corticospinal tracts. Usefulness of liver transplantation in this setting is not yet determined. We describe here the case of a 29-year-old male who presented with progressive spastic paraparesis of the lower limbs 3 years after a spleno-renal shunt. PMID- 11475144 TI - Recombinant human erythropoietin in the treatment of anemic patients with hematological malignancies. AB - Patients with malignant diseases frequently develop anemia. An alternative to blood transfusions is the application of recombinant human erythropoietin. Several nonrandomized and prospective, placebo-controlled studies have demonstrated the effect and safety of erythropoietin in patients with hematological malignancies, particularly in patients with multiple myeloma and low- to intermediate-grade non-Hodgkin's lymphoma. However, in patients with myelodysplastic syndromes, the rather low response rate of erythropoietin is overcome by the combination of erythropoietin with granulocyte colony-stimulating factor. A significant acceleration of the reconstitution of erythropoiesis has been reported in allogeneic, but not in autologous bone marrow transplantation. Especially in large open-label, multicenter studies, a statistically and clinically significant improvement in quality of life independent from chemotherapeutic response or tumor type has been demonstrated. A number of simple algorithms have been proposed using the pretreatment serum erythropoietin level, transfusion requirements, and early changes in hematological parameters. PMID- 11475145 TI - Red cell antibodies in frequently transfused patients with myelodysplastic syndrome. AB - Therapy for myelodysplastic syndrome (MDS) is often restricted to lifelong support with red blood cell units (RBCU). A variety of immune phenomena associated with antibody production have been reported in MDS patients. Therefore, we hypothesized that red cell antibodies are more frequent in patients with MDS compared to other regularly transfused patients. Red cell antibodies were determined in 42 MDS patients, in 28 patients with other hematological disorders, and in a historical group of 129 patients with end-stage renal failure. All of these patients received frequent red cell substitution therapy, at least two RBCU in biweekly intervals. Red cell antibodies were detected in 9 of 42 patients with MDS, in 3 of 28 patients with other hematological disorders, and in 4 of 129 patients with end-stage renal failure. Evidence of red cell antibodies was displayed by 6 of 27 MDS patients treated with prestorage leukocyte-depleted RBCU and 3 of 15 MDS patients transfused with bedside leukocyte-filtered RBCU. Red cell antibodies are frequent in patients with hematological disorders who require repetitive red cell transfusions. The formation of alloantibodies to red cell antigens is as frequent in MDS patients as in other patients with hematological disorders. PMID- 11475146 TI - Efficacy of fludarabine, intermittent sequential high-dose cytosine arabinoside, and mitoxantrone (FIS-HAM) salvage therapy in highly resistant acute leukemias. AB - Patients with refractory acute leukemias after intensive induction and salvage attempts have a particularly poor prognosis and therapeutic options are limited. In the current study, the pharmacologically based FIS-HAM regimen was applied, which included fludarabine 15 mg/m2 q 12 h (days 1, 2, 8, and 9), cytosine arabinoside as a 45-min infusion every 3 h at 750 mg/m2 per single application (days 1, 2, 8, and 9), and mitoxantrone 10 mg/m2 (days 3, 4, 10, 11). Twenty-six intensively pretreated patients [median age: 38 years; range: 22-65; 16 cases of acute myeloid leukemia (AML) and 10 of acute lymphoblastic leukemia (ALL)] were included. Of 16 patients with AML, 5 achieved a complete remission (CR, 31%), 1 a partial remission (PR, 6%), 2 were nonresponders (13%), and 8 succumbed to early death (ED, 50%). Of 10 patients with ALL, 5 achieved a CR, 1 a PR, 1 was a nonresponder, and 3 died early. Overall, the CR rate was 38%. The median disease free survival time was 50 days and median survival 90 days. Two patients underwent allogeneic bone marrow transplantation and are alive after 27 and 28 months. Neutropenia amounted to a median of 46 days. Toxicity WHO III/IV included infection (61%), diarrhea (48%), nausea/vomiting (43%), impairment of heart function (30%), and mucositis (26%). The current data indicate a significant activity of FIS-HAM chemotherapy in advanced acute leukemias. However, due to its pronounced toxicity, this regimen should be restricted to third-line therapy for patients expecting a suitable donor for allogeneic transplantation, and supportive treatment should be optimized. PMID- 11475147 TI - Histological subtypes of Hodgkin's disease in the setting of HIV infection. AB - The relative incidence of Hodgkin's disease (HD) has been found to have increased approximately seven times in HIV-infected patients. We analyzed the histological distribution of HIV-associated HD with the aim of clarifying purported difference(s) from de novo HD. References on HIV/AIDS-associated HD were retrieved from the most complete databases. Nineteen articles were the subject of our analysis. Seventeen of them reported data on the histological type of HIV/AIDS-associated HD patients; the route of infection and age of the patients were also considered when available. According to the Peto's methodology, histological types were compared with those from two large studies in the United States on de novo HD: 3,245 cases from the Surveillance, Epidemiology, and End Results (SEER) and 1,140 from Stanford University. The analysis of the two groups showed statistically significant differences (p<0.001) in the percentage of all histological types and odds ratios (OR) of the pooled effect of 0.4 (95% CI: 0.3 0.6) for lymphocyte predominance (LP), 0.3 (95% CI: 0.2-0.4) for nodular sclerosis (NS), 3.2 (95% CI: 2.6-3.8) for mixed cellularity (MC), and 6.3 (95% CI: 4.5-8.8) for lymphocyte depletion (LD). Comparison with the Stanford University series yielded similar results. Whilst retrospective and based on a limited number of cases, our data confirm a higher incidence of unfavorable histological subtypes in HIV-infected patients and show a reduction in the observed cases of good prognosis subtypes. Prospective studies, with careful histological observations, are required to better evaluate the characteristics of the LP subtype in the special setting of HIV infection. PMID- 11475148 TI - Deletion of chromosome 13q14 detected by fluorescence in situ hybridization has prognostic impact on survival after high-dose therapy in patients with multiple myeloma. AB - Interphase cytogenetic analysis of chromosome 13q14 was performed in 28 patients with multiple myeloma (MM) receiving high-dose therapy followed by autologous (n=24) or allogeneic (n=4) stem cell support. Eleven (39%) patients were found to have a deletion of chromosome 13q14. Response rates to high-dose therapy were independent of the chromosome 13 status, but patients with a deletion of 13q14 had a significantly shorter progression-free (p=0.001) and overall survival (p=0.012) than patients with normal chromosome 13q14. Our results indicate that high-dose therapy appears promising in patients with normal chromosome 13, whereas in patients with a deletion of 13q14 innovative therapeutic concepts are warranted. PMID- 11475149 TI - Value of Tc-99m sestamibi scintigraphy in the detection of bone lesions in multiple myeloma: comparison with Tc-99m methylene diphosphonate. AB - Technetium 99m-2-methoxyisobutylisonitrile (Tc-99m MIBI) is a lipophilic agent that accumulates preferentially within living malignant cells due to the higher transmembrane electrical potential as a consequence of the higher metabolic rate than in the surrounding normal cells. It has been effectively used to detect malignant tumors at diagnosis and follow-up and has been reported to be useful in detecting disease lesions in multiple myeloma. We studied 28 consecutive patients with multiple myeloma at diagnosis to determine the value of Tc-99m MIBI in comparison with Tc-99m methylene diphosphonate (MDP), conventional X-rays, computed tomography (CT) scan, and magnetic resonance imaging (MRI). We found 26 patients with obvious osteolytic lesions in X-rays, 22 patients with positive Tc 99m MIBI scans, and 15 patients with positive Tc-99m MDP scans. There was no coincidence of the positive lesions in the two scans, while in two patients the osteolytic areas were positive in the Tc-99m MDP scans, and in one case the osteolytic area was positive in the Tc-99m MIBI scan. The intensity of Tc-99m MIBI scans correlated with disease activity as determined by lactate dehydrogenase (LDH) (p<0.05), C-reactive protein (CRP) (p<0.01), beta2 microglobulin (p<0.05), and serum ferritin (p<0.01). We believe that Tc-99m MIBI scintigraphy can detect bone marrow lesions in myeloma patients that cannot be detected by other imaging methods and that it can be useful especially in solitary myeloma to exclude other involved sites. In addition, it could be a prognostic factor related to disease activity and multidrug resistance. We believe that a multicenter study is needed to evaluate the usefulness of this agent. PMID- 11475150 TI - Type 2B von Willebrand's disease due to Val1316Met mutation. Heterogeneity in the same sibship. AB - An analysis was conducted in four members of the same family, two of whom had a history of severe bleeding associated with type 2B von Willebrand's disease (VWD) which, although found to be due to the same mutation, nevertheless exhibited different phenotype patterns in the two subjects involved. Von Willebrand's factor (VWF) multimers were assayed with high- and low-resolution sodium dodecyl sulfate (SDS) agarose gels. The patients were studied before and after intravenous administration of desmopressin (DDAVP) at doses of 0.4 microg/kg body weight. Automatic sequencing techniques were used to analyze VWF gene exon 28. The propositus presented with mild basal thrombocytopenia with ristocetin-induced platelet aggregation (RIPA) at low concentrations of ristocetin. He had a very prolonged bleeding time (BT), and his plasma VWF was found to be lacking in large and intermediate multimers. Thrombocytopenia was observed to intensify transiently after the administration of DDAVP. The propositus' mother, in contrast, presented reduced RIPA while in a basal state, with only partial loss of the high molecular weight VWF multimers. Although she had a very prolonged BT, her platelet count was borderline. Transient correction of BT and a decrease in the platelet count were observed after administration of DDAVP and RIPA was observed at low concentrations of ristocetin. Exon 28 sequencing revealed a G4196A-->Val1316Met mutation in both patients. No other abnormality was detected within this exon. Val1316Met has been reported in type 2B VWD. In conclusion, in the family presented here, the phenotype pattern in one patient was typical of type 2B VWD, whereas the pattern in his mother was closer to type 2A VWD. After administration of DDAVP, however, a type 2B phenotype could be clearly attributed to both, indicating that this drug can be a useful tool for elucidating ambiguous phenotypes. PMID- 11475151 TI - Virus-associated hemophagocytic syndrome due to rubella virus and varicella zoster virus dual infection in patient with adult idiopathic thrombocytopenic purpura. AB - A 26-year-old woman with idiopathic thrombocytopenic purpura (ITP) was admitted to our hospital because of fever and rash. Blood tests revealed thrombocytopenia, liver dysfunction, coagulopathy, and hyperferritinemia. Bone marrow examination revealed many atypical lymphocytes and some histiocytes with hemophagocytosis. On admission she was diagnosed with rubella virus-associated hemophagocytic syndrome (VHAS), but on laboratory examination, she was seropositive for varicella-zoster virus (VZV)-IgM as well as rubella virus-IgM. She was therefore diagnosed with dual infection by rubella virus and VZV. Her simultaneous rubella virus and VZV infection may have been related to the VAHS pathogenesis. She was treated with prednisolone and gamma globulin therapy and recovered completely. PMID- 11475152 TI - Hemoglobinopathy York [beta146 (HC3) His==>Pro]: first report of a family history. AB - The rare hemoglobinopathies with abnormal oxygen binding are usually characterized by erythropoietin-mediated erythrocytosis. Bare et al. first described a hemoglobinopathy with mild erythrocytosis in a 22-year-old Caucasian woman in 1976. These authors called the abnormal hemoglobin Hb York. Hb York is characterized by a mutation at the beta146 position that changes histidine into proline. A second case of Hb York was observed by Kosugi et al. in 1983. To the best of our knowledge, no further cases have been reported. We have encountered a new case of Hb York, which was detected by agar gel electrophoresis at pH 6.0. Analysis of DNA sequences revealed a CAC-->CCC mutation in codon 146. The proportion of Hb York was approximately 50%. Analysis of oxygen transport function showed a leftward shift of the sigmoidal O2-dissociation curve. P50 was reduced to 15.5 mmHg. Investigation of family members revealed Hb York in the patient's sister, two daughters and a grandson. In retrospect, the mother of the patient may also have been affected. The mode of inheritance is autosomal dominant. PMID- 11475153 TI - Familial occurrence of the May-Hegglin anomaly: is the accompanying renal failure part of a new subentity? AB - We present two cases of the May-Hegglin anomaly discovered in a patient and one of her two sons. The female patient was known to have proteinuria from the age of 14 and was hospitalized in 1980, at the age of 25 years, because of hypertension and proteinuria (1.5 g/day). Thrombocytopenia was found with an abundance of megakaryocytes in the bone marrow. Both steroid treatment and splenectomy failed to ameliorate the thrombocytopenia, thought to be due to idiopathic thrombocytopenic purpura. Progressive renal failure, secondary hyperparathyroidism and uremic osteodystrophy were diagnosed in 1995. In January 1996, when she was hospitalized because of high-grade fever, we saw giant platelets and prominent blue inclusion bodies in almost all granulocytes in the peripheral blood smear. Electron microscopy confirmed the diagnosis of May Hegglin anomaly in this patient and one of her sons, who at that time showed thrombocytopenia but no renal disease. Three years later, however, at the age of 15, the affected son was found to develop proteinuria. Coexpression of the May Hegglin anomaly and renal disease, reported previously in a few other patients, may in fact represent a new subentity. PMID- 11475154 TI - CD20-positive T cell leukemia/lymphoma: case report and review of the literature. AB - We report on a case of CD20-positive peripheral T cell lymphoma. The lymphoma cell was positive for CD20 and T cell lineage markers such as cytoplasmic CD3, CD4, and CD5 and had a monoclonal rearrangement of the T cell receptor (TCR) gamma chain gene. The clinical characteristics resembled angioimmunoblastic lymphadenopathy: spontaneous regression of lymphadenopathy and immunological abnormalities such as polyclonal hypergammaglobulinemia, positive results of direct and indirect antiglobulin tests, and a high antinuclear antibody titer. We reviewed seven cases of CD20-positive T cell malignancies including the present case. Three were immature T cell malignancies (acute lymphoblastic leukemia) and four were peripheral T cell malignancies (non-Hodgkin's lymphoma and chronic lymphocytic leukemia). Hepatomegaly and/or splenomegaly were common features. Further cases must be evaluated to understand the clinical significance of the CD20 expression on the surface of T cell malignancies. PMID- 11475155 TI - Peripheral T-cell lymphoma in a patient with osteopetrosis. AB - Osteopetrosis is a rare genetic disorder in which the function of osteoclasts is defective, resulting in impaired bone resorption. This disease is usually accompanied by myelosuppression due to decreased marrow space and by osteomyelitis, especially in the submandibular bone. We report the case of a 72 year-old woman with an autosomal dominant form of osteopetrosis who suffered from peripheral T-cell lymphoma. Accurate clinical and pathological diagnoses and staging were difficult due to nonspecific reactive hyperplasia of the lymph nodes, even though we used several scintigraphic techniques and [18F]fluorodeoxyglucose positron emission tomography. We also paid special attention to myelosuppression and exacerbation of osteomyelitis after combination chemotherapy. Severe infectious complications were avoided by early administration of G-CSF for leukocytopenia and by continuous oral administration of antibiotics. The patient achieved complete remission after four courses of chemotherapy and this status has been maintained for 6 months. PMID- 11475156 TI - Prediction of mass-transfer coefficient for solute transport in porous media. AB - Several previously reported laboratory studies related to transport of solutes through packed columns were utilized to develop predictive relationships for mass transfer rate coefficient. The data were classified into two groups: those obtained under rate-limited mass transfer between mobile and immobile water regions (physical nonequilibrium conditions), and those derived from rate-limited mass transfer between instantaneous and slow sorption sites (sorption nonequilibrium conditions). The mass-transfer coefficient in all these studies was obtained by fitting breakthrough data to a transport model employing a first order rate limitations with a "constant" mass-transfer coefficient, independent of flow conditions. This study demonstrated that the mass-transfer coefficient in these models is dependent on system parameters including pore-water velocity, length-scale, retardation coefficient, and particle or aggregate size. Predictive relationships were developed, through regression analysis, relating mass-transfer coefficient to residence time. The developed relationships adequately estimated previously reported field mass-transfer values. Successful simulations of field desorption data reported by Bahr [J. Contam. Hydrol. 4 (1989) 205] further demonstrate the potential applicability of the developed relationships. PMID- 11475157 TI - Effectiveness of equilibrium and physical non-equilibrium approaches for interpreting solute transport through undisturbed soil columns. AB - This study observes the dispersion process of chloride through three undisturbed soil columns (sabA, sabB and arg) of different texture under both saturated and unsaturated conditions. Overall, 17 leaching experiments were conducted by means of an equipment designed and built in Portici together with the DITEC of the University of Basilicata. During the experiments, both symmetric and positive asymmetric breakthrough curves of the chloride anion were measured. As expected, manifest asymmetry was obtained especially for soil columns labeled as sabA and arg. In such columns, it was also noted that the degree of asymmetry increased as the flow density increased at the sample surface. The experiments conducted on soil column labeled as sabB resulted in more symmetric BTCs. The experimental data were firstly analysed using the classical two-parameter CD model. The variation of the dispersion coefficient, D, with pore water velocity, upsilon 0, and porous medium characteristics was explored in detail. The application to all experimental data of the four-parameter MIM model, which compartmentalises the pore water in the mobile-stagnant domains provided an insignificant different description of BTCs, even of the asymmetric ones. Furthermore, it resulted in ill defined parameters, especially those related to the partitioning in the mobile immobile phases. However, for all the three soils examined the two parameter CD model proved to be very powerful even in interpreting asymmetric BTCs. In any case, it should be emphasised that the agreement of outflow experimental data with a mechanistic model does not unequivocally identify the mechanism of solute transport in the soil. In fact, when we applied the CLT model, which is based on a stochastic approach, the BTCs were comparably well predicted. PMID- 11475158 TI - Factors affecting the distribution of hydrocarbon contaminants and hydrogeochemical parameters in a shallow sand aquifer. AB - The distributions of hydrocarbon contaminants and hydrogeochemical parameters were investigated in a shallow sand aquifer highly contaminated with petroleum hydrocarbons leaked from solvent storage tanks. For these purposes, a variety of field investigations and studies were performed, which included installation of over 100 groundwater monitoring wells and piezometers at various depths, soil logging and analyses during well and piezometer installation, chemical analysis of groundwater, pump tests, and slug tests. Continuous water level monitoring at three selected wells using automatic data-logger and manual measuring at other wells were also conducted. Based on analyses of the various investigations and tests, a number of factors were identified to explain the distribution of the hydrocarbon contaminants and hydrogeochemical parameters. These factors include indigenous biodegradation, hydrostratigraphy, preliminary pump-and-treat remedy, recharge by rainfall, and subsequent water level fluctuation. The permeable sandy layer, in which the mean water table elevation is maintained, provided a dominant pathway for contaminant transport. The preliminary pump-and-treat action accelerated the movement of the hydrocarbon contaminants and affected the redox evolution pattern. Seasonal recharge by rain, together with indigenous biodegradation, played an important role in the natural attenuation of the petroleum hydrocarbons via mixing/dilution and biodegradation. The water level fluctuations redistributed the hydrocarbon contaminants by partitioning them into the soil and groundwater. The identified factors are not independent but closely inter-correlated. PMID- 11475159 TI - Removal of volatile and semivolatile organic contamination from soil by air and steam flushing. AB - A soil core, obtained from a contaminated field site, contaminated with a mixture of volatile and semivolatile organic compounds (VOC and SVOC) was subjected to air and steam flushing. Removal rates of volatile and semivolatile organic compounds were monitored during flushing. Air flushing removed a significant portion of the VOC present in the soil, but a significant decline in removal rate occurred due to decreasing VOC concentrations in the soil gas phase. Application of steam flushing after air flushing produced a significant increase in contaminant removal rate for the first 4 to 5 pore volumes of steam condensate. Subsequently, contaminant concentrations decreased slowly with additional pore volumes of steam flushing. The passage of a steam volume corresponding to 11 pore volumes of steam condensate reduced the total VOC concentration in the soil gas (at 20 degrees C) by a factor of 20 to 0.07 mg/l. The corresponding total SVOC concentration in the condensate declined from 11 to 3 mg/l. Declines in contaminant removal rates during both air and steam flushing indicated rate limited removal consistent with the persistence of a residual organic phase, rate limited desorption, or channeling. Pressure gradients were much higher for steam flushing than for air flushing. The magnitude of the pressure gradients encountered during steam flushing for this soil indicates that, in addition to rate-limited contaminant removal, the soil permeability (2.1 x 10(-9) cm2) would be a limiting factor in the effectiveness of steam flushing. PMID- 11475160 TI - Sediment filled fractures in the Permo-Triassic sandstones of the Cheshire basin: observations and implications for pollutant transport. AB - Fracture mapping in a tunnel system and at nearby outcrop on the Runcorn Penninsula, UK, suggests the need for a review of the potential pathways for pollutant transport in Permo-Triassic sandstone aquifers. Sediment infilling is pervasive in the largest sub-vertical multi-layer fractures in the study area, both at the surface and to a depth of about 40 m below ground level. Sediment infill is inferred to have formed in situ. The conventional models of pollutant transport in fracture networks assume that they comprise open fractures, with pollutant mobility depending on fracture connectivity (a function of density, length, orientation and intersection) and aperture. The presence of extensive sediment fills in fractures will materially change their permeability, thereby reducing pollutant flux, and be of significance in the assessment of risks arising from chemical spillages. There has been little or no substantive evidence for such fills in Permo-Triassic sandstones in the UK, apart from observations at outcrop and anecdotes of sand being pumped from boreholes. Here, we report surface and rare, but complementary, subsurface observations of extensive fills in the Cheshire basin, and argue that they will only act as preferential pathways where they crosscut low-permeability horizons such as mudstones. PMID- 11475161 TI - A set of constitutive relationships accounting for residual NAPL in the unsaturated zone. AB - Although laboratory experiments show that non-aqueous phase liquid (NAPL) is retained in the unsaturated zone, no existing multiphase flow model has been developed to account for residual NAPL after NAPL drainage in the unsaturated zone. We developed a static constitutive set of saturation-capillary pressure relationships for water, NAPL and air that accounts for both this residual NAPL and entrapped NAPL. The set of constitutive relationships is formulated similarly to the set of scaled relationships that is frequently applied in continuum models. The new set consists of three fluid-phase systems: a three-phase system and a two-phase system, that both comply with the original constitutive model, and a newly introduced residual NAPL system. The new system can be added relatively easily to the original two- and three-phase systems. Entrapment is included in the model. The constitutive relationships of the non-drainable residual NAPL system are based on qualitative fluid behavior derived from a pore scale model. The pore scale model reveals that the amount of residual NAPL depends on the spreading coefficient and the water saturation. Furthermore, residual NAPL is history-dependent. At the continuum scale, a critical NAPL pressure head defines the transition from free, mobile NAPL to residual NAPL. Although the Pc-S relationships for water and total liquid are not independent in case of residual NAPL, two two-phase Pc-S relations can represent a three-phase residual system of Pc-S relations. A newly introduced parameter, referred to as the residual oil pressure head, reflects the mutual dependency of water and oil. Example calculations show consistent behavior of the constitutive model. Entrapment and retention in the unsaturated zone cooperate to retain NAPL. Moreover, the results of our constitutive model are in agreement with experimental observations. PMID- 11475162 TI - In-situ oxidation of trichloroethene by permanganate: effects on porous medium hydraulic properties. AB - In-situ oxidation of dense nonaqueous-phase liquids (DNAPLs) by strong oxidants such as potassium permanganate (KMnO4) has been proposed as a possible DNAPL remediation strategy. In this study, we investigated the effects of in-situ trichloroethene (TCE) oxidation by KMnO4 on porous medium hydraulic properties. In particular, we wanted to determine the overall effects of concurrent solid phase (MnO2) precipitation, gas (CO2) evolution and TCE dissolution resulting from the oxidation reaction on the porous medium's aqueous-phase relative permeability, krw. Three TCE removal experiments were conducted in a 95-cm long, 5.1-cm i.d. glass column, which was homogeneously packed with well-characterized 30/40-mesh silica sand. TCE was emplaced in the sand-pack in residual, entrapped form through a sequence of water/TCE imbibition and drainage steps. The column was then flushed under constant aqueous flux conditions for up to 104 h with either deionized water (reference experiment), deionized water containing 5 mM KMnO4 or deionized water containing 5 mM KMnO4 and 300 mM Na2HPO4. Aqueous-phase relative permeabilities were computed from measured flow rates and measurements of aqueous-phase pressure head, h obtained using pressure transducers connected to tensiometers distributed along the column length. A dual-energy gamma radiation system was used to monitor changes in fluid saturation that occurred during each experiment. In addition, column effluent samples were collected for chemical analyses. Dissolution of TCE during deionized water flushing led to an increase in krw by approximately 22% and a local reduction in h. On the other hand, vigorous CO2 gas production and precipitation of MnO2 was visually observed during flushing with deionized water that contained 5 mM KMnO4. As a consequence, krw declined by approximately 96% and h increased locally by more than 1000 cm H2O during the first 24 h of the experiment, causing sand-pack ruptures and pump failure. Conversely, less CO2 gas production and MnO2 precipitation was visually observed during flushing with deionized water that contained 5 mM KMnO4 and 300 mM Na2HPO4. Consequently, only small increases in h (< 15 cm H2O) were observed in this experiment due to a reduction in krw of approximately 53%. While we must attribute changes in h due to variations in krw to our specific experimental design (constant aqueous flux, one-dimensional flow experiments), these experiments nevertheless confirm that successful application of in situ chemical oxidation of TCE requires consideration of detrimental processes such as MnO2 precipitation and CO2 gas formation. In addition, our results indicate that utilization of a buffered oxidant solution may improve the effectiveness of in situ oxidation of TCE by KMnO4 in otherwise weakly buffered porous media. PMID- 11475163 TI - Factors affecting mass transfer limited biodegradation in saturated porous media. AB - Microbial degradation rates in the subsurface are not only limited by the physiological capacity of the organisms, but also by inefficient supply of nutrients to the microbes. Although mass transfer limitation of biodegradation in the subsurface has been postulated for years, experimental evidence is still scarce. In the column experiments described here, diffusive transport of 4 nitroanisole from the bulk solution to cells of Rhodococcus opacus strain AS2 immobilized on glass beads or sand appeared to be responsible for the slow transformation rates observed. Assuming steady state, we applied a coupled transformation/transport equation to these data (Best equation) and apparent bead related mass transfer coefficients were found to increase in proportion to the surface area covered with bacteria. This implies that mass transfer coefficients for individual cells remained constant. In an idealized oligotrophic environment where cells are only loosely clustered and do not shield each other, we would therefore expect biodegradation rates to be independent from the longitudinal distribution of the total biomass along a given flow path. Moreover, apparent mass transfer coefficients increased with the grain size of the column fillings, but did not change upon varying the flow rate. With a limiting external transport step, overall transformation fluxes do not become saturated at concentrations as low as predicted for Michaelis-Menten-type kinetics. Mass transfer limitation thus offers a justification for the common assumption that biodegradation rates in the subsurface follow first order kinetics in a wide concentration range. PMID- 11475164 TI - Congenital syphilis--United States, 2000. AB - In 1998, CDC initiated intensive efforts to eliminate syphilis from the United States. The following year, the National Syphilis Elimination Plan was launched with the goal of reducing primary and secondary (P&S) syphilis in adults to <0.4 cases per 100,000 population. A decrease in syphilis among women of reproductive age usually is followed by reductions in congenital syphilis (CS) rates. CS occurs when the spirochete Treponema pallidum is transmitted from a pregnant woman with syphilis to her fetus. Untreated syphilis during pregnancy may lead to stillbirth, neonatal death, and infant disorders such as deafness, neurologic impairment, and bone deformities. One of the national health objectives for 2000 was to reduce the rate of CS to <40 cases per 100,000 live-born infants. To evaluate progress toward this goal, the CS rate for 2000 was compared with the rate for 1997, the year before syphilis elimination efforts began. This report summarizes 1997-2000 CS surveillance data, which indicate that CS rates have decreased substantially among most racial/ethnic minority populations and that the elimination of CS in the United States is feasible because of the limited number of cases and highly focal distribution. To increase the percentage of women at risk who receive screening for syphilis during pregnancy, collaborative efforts are needed among health-care providers, health insurers, policymakers, and the public. PMID- 11475165 TI - Prevalence of hepatitis C virus infection among clients of HIV counseling and testing sites--Connecticut, 1999. AB - Hepatitis C virus (HCV) is a common chronic bloodborne virus infection that affects an estimated 2.7 million persons in the United States. HCV infection causes an estimated 8,000-10,000 deaths each year from cirrhosis and hepatocellular carcinoma and is the leading reason for liver transplantation. Because injection drug use is a major risk factor for both human immunodeficiency virus (HIV) and HCV transmission, publicly funded HIV counseling and testing sites (HIV CTS) may have a role in HCV prevention. To evaluate the need for HCV services at these sites, the Connecticut Department of Public Health (CDPH) conducted an anonymous HCV seroprevalence study among clients of HIV CTS. This report summarizes the results of this analysis, which indicate that, among clients of these HIV CTS, the prevalence of antibody to HCV (anti-HCV) was 9.8%, compared with 1.3% for HIV, with significantly higher prevalence among clients of substance abuse treatment sites (40.2%), compared with other sites (6.9%). HCV counseling and testing should be integrated into all HIV CTS, especially those associated with substance abuse treatment. PMID- 11475166 TI - Delayed influenza vaccine availability for 2001-02 season and supplemental recommendations of the Advisory Committee on Immunization Practices. AB - Manufacturer projections of vaccine distribution for the 2001-02 influenza season suggest that 49.8 million doses will be available for delivery by the end of October 2001; this is approximately 26 million fewer doses of influenza vaccine than were available by the end of October 1999 (75.8 million doses) (Figure 1). Manufacturers also project distribution of 27.3 million doses in November and December, bringing the cumulative projected total to 77.1 million doses, which is greater than in 2000 (70.4) and comparable with 1999 (76.8). Predictions of monthly vaccine distribution vary by manufacturer, and providers will probably receive vaccine on different schedule. PMID- 11475167 TI - Origins and treatment of airway inflammation in childhood asthma. AB - Several early events and risk factors are associated with the development of childhood asthma. Two significant risk factors are viral lower respiratory tract infections and atopy. Studies suggest that imbalances in TH1/TH2 cytokine responses in relationship to viral infections may play a role in the development of the childhood asthmatic phenotype. Airway inflammation is now recognized to contribute to the inception, persistence, and severity of asthmatic symptoms. The majority of information pertaining to airway inflammation in asthma has been derived from adult studies, but recent evaluations have been done in children. Available data are inconclusive as to the right medication to be used at the inception and during the evolution of the asthmatic phenotype in children. Inhaled corticosteroids (ICS) are not consistently effective in young children for a variety of reasons, including underlying pathophysiologic mechanisms that are unresponsive to the pharmacologic properties of ICS. The leukotriene receptor antagonists (LTRAs), recently approved for children as young as 2 years of age, address the relationship between leukotriene production and airway inflammation or remodeling in asthma. Therapeutic trials using LTRAs in children should prove beneficial. PMID- 11475168 TI - Behavioral issues in asthma management. AB - Despite advances in drug treatment, outcomes in asthma remain unsatisfactory. Often overlooked in developing treatment strategies are important factors that affect outcomes in asthma, namely behavioral issues such as the low rate of patient adherence to the prescribed treatment regimen, inadequate physician patient communication, and inconsistent implementation of evidence-based treatment guidelines. The low adherence rates among asthma patients may be improved by education programs that emphasize both the potentially serious risks of this disease and a team approach to treatment that includes patients and their families in developing a treatment plan. Interactive physician education programs have been shown to improve guideline implementation and physician communication skills. Indeed, effective physician-patient communication may be the key to improving guideline implementation and patient adherence to treatment, resulting in meaningful decreases in asthma-related morbidity and mortality. PMID- 11475169 TI - Inflammation and remodeling in the adult and child with asthma. AB - Inflammation and remodeling are characteristic features of the conducting airways in asthma, but the relationships between inflammation, inappropriate remodeling, bronchial hyperresponsiveness, and reduced pulmonary function are still unclear. In both adults and children with asthma, there are structural changes of the conducting airways that include injury and loss of the surface epithelium, thickening of the reticular basement membrane, increases of underlying collagen, blood vessels, and airway smooth muscle, and plugging of the airways by exudate. Bronchial biopsies obtained from persons with mild stable asthma already demonstrate the presence of inflammation. Many of the inflammatory and structural changes begin early in childhood. Whereas corticosteroids markedly reduce many aspects of inflammation, it is not known whether and how they affect the changes associated with airway wall remodeling. Leukotriene receptor antagonists appear to be antiinflammatory and able to reduce the proliferation of bronchial smooth muscle. PMID- 11475170 TI - Pediatric asthma: "Real world" measures of effectiveness. AB - In the treatment of pediatric asthma, measuring the effectiveness of an intervention in terms of cost or therapeutic value can be a challenging process. Parameters that should be considered in determining the cost effectiveness of an intervention vary considerably. In clinical therapeutic trials of pediatric asthma, subtle patient selection biases, difficulties in choosing and assessing treatment outcomes in young children, and the highly variable nature of asthma symptomatology can compromise valid measurements of the therapeutic effectiveness of an intervention. Ideally, outcome measurements in asthma should reflect "real world" conditions. They should be as broad-based as possible and include both objective measures of pulmonary function and subjective measures of quality-of life evaluated over the long term. Patient adherence is thus a critical variable that influences the ultimate effectiveness of treatment. The rate of patient adherence is influenced not only by drug characteristics such as route of administration and dosing frequency, but also by the impact of treatment on a patient's quality of life. Because asthma is a chronic disease with variable symptomatology, clinical trials investigating the cost and therapeutic effectiveness of asthma interventions must evaluate real-world variables in long term trials. This article will provide information regarding proper assessment of an intervention's cost and therapeutic value. PMID- 11475171 TI - Controversies and challenges of exercise-induced bronchoconstriction and their implications for children. AB - The association of exercise with hyperresponsiveness and bronchoconstriction of the airways has been well-recognized. Exercise may be the sole trigger or one of many triggers in patients with asthma. Although it is clear that asthma and the symptoms brought about by exercise are closely intertwined, the pathophysiology underlying this process is not well-understood. Recent studies on the epidemiology of the disease have provided insights into the processes leading to exercise-induced bronchoconstriction, with particular significance for children with asthma. PMID- 11475172 TI - Montelukast in 2- to 5-year-old children with asthma. AB - Pediatric dose selection for montelukast in 2-5-year-old children was based on a pharmacokinetic approach that was also used in previously selecting a once-daily 5-mg chewable tablet for asthmatic children 6-14 years of age. A 4-mg chewable tablet administered once daily was determined to be the correct dose and was subsequently found to be safe and effective in asthmatic children 2-5 years of age. PMID- 11475173 TI - Issues in pediatric asthma. AB - Different wheezing syndromes can carry the diagnostic label of "asthma," especially in very young children, and an accurate differential diagnosis is essential for improving outcomes. Because presenting symptoms are similar, making a rapid, accurate differential diagnosis is often daunting. Asthma can, eventually, be distinguished from other wheezing disorders based on differences in symptomatology, pathophysiology, and disease evolution. Once diagnosed, inhaled corticosteroids (ICS) remain the cornerstone of treatment in persistent pediatric asthma. Compelling evidence supports the benefits of these agents in reducing asthma-related morbidity and mortality and in preventing airway remodeling. As a result, ICS use in young children with asthma has grown substantially in recent years. Questions are being raised about the appropriateness of early and long-term use of ICS in young children, and whether ICS are overused in this population. Because of the challenges faced by clinicians in differentiating asthma from the other childhood wheezing disorders often present in young children, it may be best to delay the use of regular ICS therapy until a definitive diagnosis can be established. Alternatives to ICS (such as the leukotriene receptor antagonists) should also be considered to avoid the growth-suppressing potential of steroids in the management of mild asthma. PMID- 11475174 TI - Facilitation of fiberoptic nasotracheal intubation by simultaneous direct laryngoscopy in anesthetized patients. AB - Two cases of unexpected difficult intubation during induction of general anesthesia were reported. The first case, a 68-year-old male was scheduled for coronary artery bypass surgery. The second case, a 94-year-old male with senile dementia was arranged for exploratory laparotomy. Anesthesia was induced with diazepam 10 mg, fentanyl 30 micrograms/kg, and pancuronium 8 mg in the first case while in the second case fentanyl 100 micrograms, lidocaine 80 mg, thiopental 200 mg and succinylcholine 80 mg were used. In these two cases oral tracheal intubation with laryngoscope was unsuccessful. Assisted ventilation could be maintained via a face mask. With the help of direct laryngoscopy, fiberoptic bronchoscope-aided nasotracheal intubation was successfully achieved. PMID- 11475175 TI - Is it a dream--the possibility of development of a new venous air embolism monitor in the near future? PMID- 11475176 TI - Oxygen in nitrogen versus nitrous oxide during pediatric general anesthesia. AB - BACKGROUND: Induction of anesthesia decreases lung volumes, giving areas of non ventilated lung. Nitrogen is a slowly-absorbed gas that could prevent resorption of gases from these closed spaces, preventing atelectasis and improving oxygen saturations during recovery. METHODS: We evaluated oxygen saturations during emergence after intra-operative administration of 33% oxygen in nitrogen versus 33% oxygen in nitrous oxide in 62 children having elective urologic surgery. Patients were randomly assigned to either group A (2.5 L/min air with 0.5 L/min oxygen) or group N (2 L/min nitrous oxide and 1 L/min oxygen). Flowmeters were covered to maintain blinding. Anesthetic technique was standardized (laryngeal mask airway, caudal, halothane, and deep extubation). Patients breathed room air during emergence, while a blinded observer recorded duration of desaturations by stopwatch for 15 min. RESULTS: We found similar desaturations in both groups. The difference in desaturations < 94% at an interim 2-min total probably reflects diffusion hypoxia and was not significant at 15 min. The frequency of desaturations < 87% during emergence was significantly greater in children who were crying during induction. CONCLUSIONS: The intra-operative use of air versus nitrous oxide has no substantial effect on oxygen saturations during emergence from anesthesia in children. These results are consistent with a recently published mathematical model. PMID- 11475177 TI - The effect of intraischemic mild hypothermia on focal cerebral ischemia/reperfusion injury. AB - BACKGROUND: The protective role of intraischemic mild hypothermia on brain injury associated with cerebral ischemia/reperfusion was investigated using a localized, no-flow cerebral ischemia/reperfusion model in rats in vivo. METHODS: Male Wistar rats weighing 250-350 g were anesthetized lightly with halothane. All rats were randomly allocated into two series experiments the first of which was for study of infarct volume and the second of which was for study of NO (nitric oxide) content, and then subjected to various time intervals of focal ischemia followed by reperfusion. The intraischemic mild hypothermia was induced by feedback regulated semiconductor cooling block in a group of these rats. Normothermia groups were considered as the control groups. The infarct volume and the NO content of rat brain were measured in different groups and hours respectively. RESULTS: The infarct volume of mild hypothermia group was significantly different from that of normal control group within the time window of 10 min (P < 0.05), infarcted brain volumes in hypothermic rats were significantly smaller than in normothermic rats, for the time window of 1 h there was no significant difference in the infarct volume between mild hypothermic group and normothermic control group (P > 0.5). Furthermore, increased tissue levels of NO, a potential mediator of cerebral injury in the brain following focal ischemia were depressed by mild hypothermia. CONCLUSIONS: It is suggested that localized mild hypothermia may be a protective against focal cerebral ischemia/reperfusion injury and that the decreased nitric oxide production in the brain may serve as the protective mechanism of hypothermia. PMID- 11475178 TI - Hemodynamic changes caused by venous gas embolism in dogs: comparisons among air, carbon dioxide and oxygen. AB - BACKGROUND: The volume of a bulky venous air emboli (VAE) can be estimated based on the calibration curve generated by injections of minute amount of air into the right atrium (RA) of dogs. We speculated that in patients similar VAE calibration curves could be generated using CO2 injections. As part of pre-clinical evaluation of the usefulness of CO2 injection, the present study was designed to determine whether injection of CO2 into the RA would cause less hemodynamic changes in dogs as compared with that of air or O2. METHODS: Twenty-one anesthetized mongrel dogs were divided into 3 groups, i.e., groups air, CO2 and O2 (n = 7 each). Animals were injected a bolus of gas (air, CO2 or O2) in increasing volumes (from 0.25 to 4.0 mL/kg) into the RA via a central venous catheter at 10-min intervals. We measured the maximal changes in mean arterial pressure (MAP), pulmonary arterial pressure (PAP), central venous pressure (CVP), end-tidal CO2 (ETCO2), heart rate (HR), and mixed venous O2 saturation (SvO2). RESULTS: After venous injections, dogs receiving O2 or air showed greater decreases in MAP and ETCO2 as compared with those receiving CO2 (volume > 2.5 mL/kg; P < 0.05). The increases in PAP and CVP bore direct relation to O2 and air volume. In the CO2 group, the maximal changes in PAP and CVP were subtle as compared with the baseline (P < 0.05). There were no significant differences in HR and SvO2 among three groups. CONCLUSIONS: Injections of CO2 in increasing volumes into the RA of dogs caused less hemodynamic changes in comparison with that of air and O2. PMID- 11475179 TI - Lack of intravenous lidocaine effects on HRV changes of tracheal intubation during induction of general anesthesia. AB - BACKGROUND: Intravenous lidocaine has been widely used for suppressing the autonomic activation from tracheal intubation during induction of general anesthesia. Conventionally, researches of its effectiveness through assessment of heart rate and blood pressure changes obtained by common clinical methods result in the conclusions deduced of much controversy. Heart rate variability is a noninvasive measurement of autonomic regulation and is suitable for the study of this subject. METHODS: 36 ASA class I-II patients undergoing general anesthesia were divided into 3 groups. Besides induction agents, intravenous lidocaine was given 5 min before tracheal intubation in group A, 3 min before intubation in group B and nothing in group C. HRV spectral powers were measured at awake state, anesthetized state before tracheal intubation and anesthetized state after tracheal intubation by time frequency spectral analysis method and comparison was made between the three groups. RESULTS: The HRV spectral power in high frequency (HF) and mid-frequency (MF) power bands and their ratios (MF/HF) were not significantly different among the 3 groups during the 3 observation periods. CONCLUSIONS: There was no evidence to indicate the effectiveness of intravenous lidocaine on the autonomic regulation during tracheal intubation under the influence of induction agents used in general anesthesia. PMID- 11475180 TI - Low dose fentanyl and propofol improve the speed and quality of tidal-breathing induction techniques in sevoflurane anesthesia for adults. AB - BACKGROUND: The objective of this study was to investigate whether low dose fentanyl, with or without low dose propofol, as pretreatment agent/s is capable of speeding up and improving the quality of laryngeal mask airway (LMA) insertion in tidal-breathing induction technique with high-concentration sevoflurane. METHODS: One hundred and twenty patients were assigned to one of the three groups: Group S, induction with 8% sevoflurane only; Group F + S, 1.0 microgram/kg fentanyl prior to induction; and Group F + P + S, 1.0 microgram/kg fentanyl and 0.5 mg/kg propofol prior to induction. RESULTS: It was demonstrated that the time from administration of drug (drugs) to loss of eyelash reflex (P < 0.05, Group F + P + S vs. F + S; P < 0.01, Group F + P + S vs. S), to jaw relaxation (P < 0.05, Group F + P + S vs. S) and time taken for LMA insertion (P < 0.01, Group F + P + S vs. S) were all shorter in Group F + P + S, with fewer complications (coughing and involuntary movement) during induction, however, the first time success rate with LMA insertion did not significantly differ among the comparing groups. According to a postoperative inquiry (by questionnaire), there were significantly more patients in the Group F + P + S (57.5%) who considered the induction as pleasant (P < 0.05), of whom 75% expressed that they would be willing to undergo an induction of the same form again in the future (P < 0.05). This more positive rating may be related to the mild sedative effects of the agents given and shorter induction time, which significantly helped reduce the rate of recall of the unpleasant gas. CONCLUSIONS: The results of this study of LMA insertion, for ASA I or II adult patients undergoing the tidal-breathing technique with 8% sevoflurane, suggest that pretreatment with 1 microgram/kg fentanyl plus 0.5 mg/kg propofol is superior in comparison with either pretreatment with 1 microgram/kg fentanyl or absence of pretreatment. PMID- 11475181 TI - Pneumopericardium after bronchoscopic carbon dioxide laser surgery--a case report. AB - A female baby sustained pneumopericardium after bronchoscopic CO2 laser surgery for tracheal stenosis. Perioperative pneumopericardium though rare is a life threatening condition in babies. The symptoms are usually not specific such as hypotension, bradycardia, muffled heart sound, cardiovascular collapse, etc. The causes and clinical findings of pneumopericardium were reviewed, as are the radiographic features so as to differentiate the condition from pneumomediastinum. Early recognition of pneumopericardium is important, because emergent pericardiocentesis may be required if there is clinical evidence of cardiac tamponade. PMID- 11475182 TI - Delayed airway obstruction secondary to inadvertent arterial puncture during percutaneous central venous cannulation. AB - We report a case of fatal respiratory complication secondary to central venous cannulation in a 63-year-old male patient, which came up gradually and insidiously following an accidental puncture of carotid artery in an attempt to cannulate the right internal jugular vein. He died 14 h after the mishap due to severe upper airway obstruction. The nature of the vascular laceration was still obscure. PMID- 11475183 TI - Reperfusion pulmonary edema after pulmonary endarterectomy. AB - Pulmonary artery thromboendarterectomy is a potentially curative procedure in chronic, major vessel thromboembolic pulmonary hypertension. However, persistent pulmonary hypertension and unrelenting reperfusion edema have serious complications, often requiring prolonged mechanical ventilation. A 50-year-old man who was diagnosed with a thromboembolism in both pulmonary arteries underwent a bilateral pulmonary endarterectomy. He received O2-isoflurane-fentanyl anesthesia. When the lungs were reperfused with CPB weaning, massive hemorrhage occurred in the left lung. After the operation, the patient was taken to the intensive care unit. Mechanical ventilation was performed immediately and then both inhaled NO and i.v. furosemide therapies were administered. The patient was discharged from ICU 15 days postoperation. PMID- 11475184 TI - [The era of general practice has not gone by!]. PMID- 11475185 TI - [Selective serotonin uptake inhibitors--are severe adverse effects a problem?]. PMID- 11475186 TI - [When should a fatal case occurring in hospital be reported to the police?]. PMID- 11475188 TI - [Does the use of computerized medical records improve the treatment of patients?]. PMID- 11475187 TI - [Import of resistant bacteria]. PMID- 11475189 TI - [Blood pressure records in general practice--expectations and reality]. AB - BACKGROUND: The effect of antihypertensive medication on cardiovascular diseases is based on results achieved in prospective and controlled clinical studies. Comparable results in clinical practice can be achieved only when the quality of management for subjects with high blood pressure is comparable with the quality achieved in clinical studies. MATERIAL AND METHODS: This study consists of two parts; an attempt to implement an computer-based clinical decision support system for management of hypertension in primary health care in two counties in Norway, and a questionnaire survey among doctors in primary care and specialists in internal medicine in hospitals in other counties. We asked what they expected would be the result of the specific implementation strategy for the computer based program. The objectives were to evaluate the results of the implementation strategy and to compare these results with expectations expressed in the filled in questionnaires by doctors without any obligations to the main study. RESULTS: A total of 175 doctors were invited to implement the clinical decision support system. 85% responded; 44% of these, or 37% of the invited doctors, were willing to participate. After 12 months with recurrent visits by one of the authors, only six out of 74 doctors participating in the intervention study still used the program. The questionnaire were completed and returned by 203 doctors, who expected that 55% of the invited doctors would accept the invitation following the implementation strategy used. In general the validity of the information given by the questionnaire was poor and unreliable. INTERPRETATION: We conclude that introduction of a computer-based clinical decision support system is difficult in a busy primary care setting. Availability and simplicity are crucial requirements, and doctors would need financial compensation if they were to use such a system. PMID- 11475190 TI - [Perilymphatic fistula]. AB - BACKGROUND: Perilymphatic fistula is leakage of perilymph from the inner ear to the middle ear. The perilymph surrounds the endolymphatic system in the inner ear (cochlea and the vestibular apparatus) and is most likely a cerebrospinal fluid filtrate. Leakage can occur through the fenestra vestibuli (the oval window), the fenestra cochleae (the round window), or pathological openings in the otic capsule. The cause of perilymphatic fistula can be trauma (barotrauma, head trauma, whiplash injury, iatrogenic trauma during ear surgery or ear syringing), chronic otitis media with cholesteatoma, congenital malformations or idiopathic. MATERIAL AND METHODS: We present 15 patients operated at Haukeland University Hospital from 1980 to 1997. RESULTS: Symptoms and signs are not very consistent, but dizziness and hearing loss are usual. The fistula test has low sensitivity and specificity, and otomicroscopy is not diagnostic. Imaging techniques with CT and MRI are seldom helpful unless in cases of inner ear abnormalities. Surgical exploration of the middle ear can be diagnostic, but even then it may in some cases be difficult to determine if a perilymphatic fistula is present. INTERPRETATION: The treatment of suspected perilymphatic fistula is primarily bed rest with elevated head, and avoidance of straining to facilitate spontaneous closure of a possible fistula. If the symptoms continue, or get worse, surgical exploration is indicated. The results are good regarding dizziness, but more uncertain as far as hearing improvement is concerned. Hearing improvement seems to be negatively correlated with the length of the delay before the fistula is closed. PMID- 11475191 TI - [Salivary gland drainage into the thyroglossal duct]. AB - BACKGROUND: Failure in regression of the thyroglossal duct is one of the most common reasons for midline swellings in the neck. Several authors have described recurrent thyroglossal duct remnants with persisting draining sinuses. However, few have described accessory salivary glands that drain into the thyroglossal duct. MATERIAL AND METHODS: In this article we report two such cases with midline salivary glands in the floor of the mouth. RESULTS: These two patients were subsequently successfully treated with radical tissue resection in the area between the hyoid bone and foramen cecum. INTERPRETATION: Preoperative fistulography or sinography was useful to demonstrate the ductal ramification of the salivary glands, and use of methylene blue during surgery proved of significant value for the result. PMID- 11475192 TI - [Pneumocephalus as a complication to sinus surgery]. AB - BACKGROUND: Pneumocephalus is a rare complication of sinus surgery. The complications of sinus surgery are often classified as minor or major. Minor complications are seen as adhesions, scarring, bleeding, development of subcutaneous emphysema, penetration of the orbit without herniation and eyelid ecchymosis. The incidence of major complications is 0 to 3% and include haemorrhage, retrobulbar haematoma, injury to the optic nerve, CSF leaks, meningitis, intracranial penetration with development of tension pneumocephalus, and death. MATERIAL AND METHODS: We present a case of pneumocephalus after endonasal ethmoidectomy. The literature is reviewed. RESULTS: The patient had clinical findings consistent with non-tension pneumocephalus. Development of tension pneumocephalus is a distinct possibility and it is necessary to monitor these patients closely. INTERPRETATION: In order to reduce the risk of injury to the anterior skull base it is important to eliminate certain normal anatomic variations and to assess the individual anatomy thoroughly. This can be done with preoperative nasal endoscopy and CT imaging. This applies even more to patients who have previously undergone sinus surgery. PMID- 11475193 TI - [Takayasu's arteritis]. AB - BACKGROUND: Takayasu's arteritis is a chronic, idiopathic, inflammatory disease that affects aorta and its main branches. The disease is rare; its etiology is unknown and shows race differences. The inflammation of the arteries may lead to stenosis, occlusions, dilatations or aneurysms. The clinical picture and angiographic findings are not previously reported in a Norwegian cohort. MATERIAL AND METHODS: We report a retrospective, hospital-based study describing the clinical picture, diagnostic findings, treatment and prognosis in a cohort of six patients in Central Norway with Takayasu's arteritis. The data was extracted through chart review. RESULTS: In the period 1988-2000, six patients with Takayasu's arteritis, five women and one man, were identified. All the patients were of Norwegian origin. Median age at diagnosis was 39 years, range 24-63 years, and median time from first symptoms to definite diagnosis was six months, range 1-36 months. The estimated minimum annual incidence was 0.8 per million. All patients had elevated erythrocyte sedimentation rate; five out of six patients had unilateral or bilateral subclavian stenosis; one patient had a thoracoabdominal aneurysm. All patients were treated with prednisolone. There were no deaths in the observation period of median 7.5 years, range 0-26 years. INTERPRETATION: Takayasu's arteritis is a rare disease in our region, with lower incidence than reported in the literature. The prognosis is excellent, but the morbidity was substantial. The clinical findings are similar to those reported in other studies. The location and appearance of the angiographic findings were characteristic for the disease. PMID- 11475194 TI - [Selective serotonin uptake inhibitors and pancreatitis]. AB - BACKGROUND: About 2% of all cases of pancreatitis are drug-induced. Several drugs cause pancreatitis, including selective serotonin reuptake inhibitors (SSRIs). MATERIAL AND METHODS: Two Norwegian adverse drug event reports and a review of the international WHO adverse drug reactions database form the basis for this discussion. We found a total of 160 cases of pancreatitis associated with SSRIs. RESULTS: Our findings support the notion that SSRIs may trigger pancreatitis in rare cases. The course of the disease may be severe and even fatal. INTERPRETATION: Physicians should be aware of the possibility that SSRIs may trigger pancreatitis. Patients taking SSRIs should be evaluated for pancreatitis if they report abdominal pain of unknown cause. PMID- 11475195 TI - [Air in the portal vein and necrosis of the ventricle]. AB - BACKGROUND: Air in the portal vein is a serious sign, usually caused by enteric necrosis. METHODS: The case report presents a 69-year-old male who died shortly after arrival in our department. RESULTS: Abdominal CT scan showed hepatic portal venous gas, and thrombi in the superior mesenteric artery, the coeliac trunk and in the left iliac artery. Autopsy revealed a necrotic stomach but viable small bowel. INTERPRETATION: Differential diagnoses are discussed, and we conclude that the patient most probably died from ischaemic necrosis. In severely ill patients, hepatic portal venous gas is an indication for urgent laparotomy. PMID- 11475196 TI - [Three primary stents in extensive esophageal cancer]. AB - BACKGROUND: Most patients with oesophageal cancer have unresectable disease at initial assessment. Dysphagia is the most common symptom. Laser ablation and self expandable metal stents are among the possible treatments for dysphagia caused by oesophageal cancer. There are two types of self-expandable metal stents: covered and uncovered. Tumour can easily grow through uncovered stents. Such tumour ingrowth can be treated with laser ablation or Argon plasma coagulation. MATERIAL AND METHODS: We present a 54-year-old man with metastatic oesophageal cancer. RESULTS: Because of dysphagia, self-expandable stents were inserted into his oesophagus. At the time of stent insertion, the primary tumour was 23 cm in craniocaudal direction. We inserted three stents, each 10 cm long. The patient restored normal swallowing and had no pain, dyspepsia or unpleasant sensation from his stents. Uncovered stents were used. Tumour ingrowth was treated with Argon plasma coagulation. INTERPRETATION: In this patient, with a long tumour, the use of three primary stents reduced dysphagia. PMID- 11475197 TI - [Glycopyrrolate treatment for drooling in an adult with cerebral palsy]. AB - BACKGROUND: The aim of the study was to assess the effect of glycopyrrolate on drooling in an adult male patient with cerebral palsy. MATERIAL AND METHODS: After a thorough medical examination and consent in writing from his guardian, a baseline sum score for frequency and seriousness of drooling was established over a two-week period in addition to data on shifts of bibs, urination, defecation and observation of behaviour. Glycopyrrolate (1 mg) tablets were then administered, starting with one tablet daily the third week and increasing the daily dose by one tablet per week until a maximum of four tablets during week 6 and four days of week 7, when the daily dose was reduced to two tablets for three days. In the four weeks 8-11, three tablets were given daily. In week 12 the dose was reduced to two tablets and for the weeks 13-15 no glycopyrrolate was given. RESULTS: As long as the patient received three to four tablets of glycopyrrolate daily, drooling was markedly reduced, and bibs were not necessary on some days. After withdrawal, drooling increased to approximately the same level as before treatment. INTERPRETATION: For shorter periods, glycopyrrolate can be given in controlled doses provided that an adequate medical assessment has been undertaken. PMID- 11475198 TI - [Metabolic interactions with statins]. AB - BACKGROUND: Isoenzymes of the cytochrome P450 (CYP) system play a prominent role in drug metabolism, including the HMG-CoA reductase inhibitors (statins). Alteration in metabolic activity is often the underlying mechanism of clinically relevant interactions, thus it is important to make a rational assessment of the risk for metabolic interactions with statins. MATERIAL AND METHODS: Articles covering the field of statin metabolism and related interactions were mainly searched for via PubMed. RESULT: More than 50% of the overall CYP metabolism is mediated through the isoenzyme CYP3A4, which is the main elimination route of simvastatin, lovastatin and atorvastatin. Interaction studies reveal that simvastatin and lovastatin have the highest potential for clinically relevant interactions related to this isoenzyme, most often leading to increased efficacy/toxicity of the statin. Interactions with agents that alter the activity of CYP3A4 can also be expected for atorvastatin. Other statins are to a lesser extent dependent on CYP3A4 in their elimination. Thus, pravastatin, cerivastatin and fluvastatin have a minor potential for interactions linked to this isoenzyme. Interactions associated with CYP2C9-interference may, however, be present for fluvastatin. INTERPRETATION: Since lipid-lowering therapy most often is life long, the use of statins will frequently be accompanied by the use of multiple drugs, hence the importance of considering the risk for interactions when selecting statins. PMID- 11475199 TI - [Peripheral neuropathy in cancer]. AB - Neuropathy is a quite common finding in cancer patients. If the cancer is known, the main clinical problem is often whether the symptoms and signs are caused by metastasis or recurrence, or if the neuropathy can be related to effects of the oncological treatment. In addition to the clinical findings, radiological and neurophysiological examination can often be of considerable diagnostic help. Peripheral neuropathy may also be the first manifestation of the cancer, as in paraneoplastic neuropathies. Detecting the tumor in these cases may improve the oncological prognosis, as the neuropathy often precedes tumor symptoms by several years. A determined approach in searching for an occult cancer is necessary. The neuropathic symptoms are seldom life-threatening. However, the neuropathy may cause distressing symptoms, and symptomatic treatment and rehabilitation is often required. PMID- 11475200 TI - [The transfer of selective serotonin reuptake inhibitors to human milk]. AB - BACKGROUND: This article presents an overview of the excretion of the SSRIs citalopram, paroxetine, fluoxetine, fluvoxamine and sertraline in breast milk. MATERIAL AND METHODS: Published articles on selective serotonin reuptake inhibitors and excretion in breast milk were identified and reviewed. In addition, drug concentrations were measured in milk from eight women using paroxetine (n = 4), citalopram (n = 3) or fluvoxamine (n = 1). RESULTS: Data from the literature indicate that the relative dose to the infant is lowest for fluvoxamine and sertraline, somewhat higher for paroxetine and highest for citalopram and fluoxetine. Adverse effects were reported in three of the 119 breastfed infants. Our own results show minimal excretion of fluvoxamine, small excretion of paroxetine and higher excretion of citalopram into breast milk. INTERPRETATION: If treatment with a selective serotonin reuptake inhibitor is started during the postpartum period, fluoxetine should not be the first alternative. High doses of citalopram should also be used with caution. However, when the use of an SSRI is clearly indicated in a breastfeeding woman, available data generally indicate that the positive effects of breast-feeding outweigh the risks for pharmacological effects in the infant. PMID- 11475201 TI - [Infection outbreaks caused by methicillin-resistant Staphylococcus aureus at Haukeland hospital]. AB - BACKGROUND: Nosocomial infections caused by methicillin-resistant Staphylococcus aureus (MRSA) represent an increasing problem worldwide. MATERIAL AND METHODS: We report two outbreaks of methicillin-resistant S. aureus at Haukeland University Hospital during 1998-99. RESULTS: During the fall of 1998, four patients in the dermatological ward and three of their relatives were infected or colonised with MRSA. The strain was probably introduced by an eczematous patient who had recently arrived from Japan. Three patients became chronic carriers. The second outbreak involved three other hospital departments in July-August 1999. The index patient, a multitraumatised japanese tourist, died 16 days after admission. Two other patients were infected, one of them became a chronic carrier. According to official guidelines, neither of the index patients needed MRSA screening. Pulsed field gel-electrophoresis of the MRSA isolates revealed two different strains at the first outbreak, both probably introduced from the index patient, and one single strain at the second outbreak. Nasal swabs of staff were negative. INTERPRETATION: Physicians need to know that recommended guidelines regarding MRSA screening do not cover all types of risk situations. MRSA carriage among healthcare workers is probably not an important source of MRSA spread in hospitals. Measures to prevent cross infection between patients should be emphasised. PMID- 11475202 TI - [Fridtjof Nansen as a neurobiologist]. AB - Fridtjof Nansen (1861-1930) started his four-year research carrier in neurobiology at the Museum in Bergen in 1882. His PhD thesis from 1886 describes the structure of the nervous system in selected groups of invertebrates and in Amphioxus and the hagfish. For his time, he was a firm antireticularist and is still recognised as a forerunner of the emerging neurone doctrine of contiguity and no continuity between neurons in neural pathways. PMID- 11475203 TI - [Digitalis in heart failure--still a therapeutic principle of current interest?]. PMID- 11475204 TI - [Meeting of human beings during the consultation]. PMID- 11475205 TI - [Dismantling of the welfare state]. PMID- 11475206 TI - [In the shadow of the new laws--a fatal case]. PMID- 11475207 TI - [Is approval of a multidisciplinary course by the Medical Society inappropriate?]. PMID- 11475208 TI - [Is Alzheimer disease a nutritional problem?]. PMID- 11475209 TI - [Is hypothyroidism underdiagnosed?]. PMID- 11475210 TI - [Coordination of psychosocial work and evidence-based medicine]. PMID- 11475211 TI - [Zyban in smoking cessation--what about medical ethics?]. PMID- 11475212 TI - [My way to diagnosis]. PMID- 11475213 TI - [Elderly physicians]. PMID- 11475214 TI - [Physicians with blinkers]. PMID- 11475215 TI - [A lot can be changed for the patients with stroke]. PMID- 11475216 TI - [Primary hyperparathyroidism--an underdiagnosed disease]. PMID- 11475217 TI - [Changing treatment of diabetes]. PMID- 11475218 TI - [Genetic tests]. PMID- 11475219 TI - [Solidarity thinking with one foot in the grave]. PMID- 11475220 TI - [Should patients with acute stroke routinely receive supplemental oxygen?]. AB - BACKGROUND: Supplemental oxygen is often given routinely to all patients suffering from an acute stroke, although clinical evidence for its efficacy is not available. The object of this study was to study the impact on mortality, impairment and disability of supplemental oxygen given the first 24 hours after an acute stroke. MATERIAL AND METHODS: Patients admitted to hospital with acute stroke were randomly allocated to two groups: one group received supplemental oxygen (100% atmospheres, 3 litres/minute) for 24 hours (n = 292); whereas a control group did not receive additional oxygen (n = 258). RESULTS: One-year survival was 69% in the oxygen group and 73% in the control group (odds ratio 0.82; 95% CI 0.57-1.19; p = 0.30). Impairment scores and disability scores were comparable seven months after stroke. Among patients with Scandinavian Stroke Scale (SSS) > or = 40.82% in the oxygen group and 91% in the control group survived (odds ratio 0.45; 95% CI 0.23-0.90; p = 0.02). For patients with SSS < 40, 53% in the oxygen group and 48% in the control group survived (odds ratio 1.26; 95% CI 0.76-2.09; p = 0.54). INTERPRETATION: The study indicates that supplemental oxygen should not be given routinely to non-hypoxic stroke victims with minor or moderate strokes. Further research is needed for giving conclusive advice concerning oxygen supplementation for patients with severe strokes. PMID- 11475221 TI - [Primary hyperparathyroidism--a 16-year material from a central hospital]. AB - BACKGROUND: In Norway, population 4.3 million, approximately 160 patients a year have undergone surgery for primary hyperparathyroidism during the last ten years. Previous reports are rare. MATERIAL AND METHODS: A retrospective analysis of all patients operated for primary hyperparathyroidism during 16 years at Vestfold Hospital, which serves a population of 200,000. Late recurrences and persisting hypocalcaemia were evaluated with laboratory tests median eight years after surgery. RESULTS: 104 patients were operated (88 females and 16 males; median age 64 years, range 22-78). Adenomas were found in 88 patients, of which four had double adenomas, and hyperplasia in 16 patients. Three patients died postoperatively, within 30 days, all of them high-risk patients. Two patients were reoperated because of bleeding. In one patient an infection was drained. Median hospital stay was five days. Two patients had persisting hypercalcaemia; eight patients needed permanent vitamin D substitution. Four recurrences were detected. Normocalcaemia was achieved in 91 patients (90%). INTERPRETATION: Surgical treatment leads to normocalcemia in 90% of patient, but is not risk-free in severely ill patients. To assess the true benefit of the operation for the individual patient, better preoperative mapping of symptoms and better follow-up protocols should be implemented. PMID- 11475222 TI - [Cardiac tumors]. AB - BACKGROUND: Tumours of the heart are rare, but of great clinical interest as the majority are benign and amenable to radical excision. MATERIAL AND METHODS: We report on 30 patients with cardiac tumours resected at the National Hospital, Oslo, Norway from 1990 to 1999. Data were obtained from patient files. RESULTS: 27 patients had benign tumours, 25 of which were myxomas predominantly located in the left atrium. One patient had a primary malignant tumour (rhabdomyosarcoma) and two had secondary malignant tumours (metastases from malignant tumours). All patients were alive at follow-up four to ten years after operation. INTERPRETATION: Patient with benign tumours are cured by surgery, whereas in malignant primary tumours the surgery will most often be palliative. Surgery for cardiac metastases may be indicated in selected symptomatic patients. PMID- 11475223 TI - [Cardiac manifestations in Fabry disease]. AB - BACKGROUND: Fabry's disease is an X-linked inborn error of metabolism. The patients lack or have very low activity of the enzyme alpha-galactosidase A. This results in deposition of sphingolipids in endothelial cells and vascular smooth muscle cells; thus the disease can affect nearly every organ in the body. Renal failure is the most common cause of death, but cardiac involvement is frequent. MATERIAL AND METHODS: We describe two brothers with Fabry's disease and provide a review of the literature in the field. RESULTS: Both patients had extensive electro- and echocardiographic findings. INTERPRETATION: Fabry's disease should be suspected in men with unexplained electro- and/or echocardiographic signs of left ventricular hypertrophy and a short PQ interval in the ECG. PMID- 11475224 TI - [Esophageal lesions associated with diphosphonates]. AB - BACKGROUND: Bisphosphonates are potent inhibitors of osteoclast-mediated bone resorption and effective in preventing osteoporotic fractures, but they can occasionally cause oesophageal adverse events. MATERIAL AND METHODS: We report on seven patients who developed severe oesophagitis or oesophageal ulceration during treatment with bisphosphonates. They were registered at our endoscopy unit during a 31-month period. RESULTS: Six of the patients took alendronate (Fosamax) and one etidronate (Didronate). The oesophageal lesions heal on discontinuation of the bisphosphonate. Institution of gastric acid suppression treatment may enhance the healing process. INTERPRETATION: In order to minimise the risk of serious side-effects, it is important to give detailed instructions regarding medication and to ensure that the instructions are properly understood. The risk of serious complications can be reduced by early recognition of oesophageal symptoms and appropriate intervention. PMID- 11475225 TI - [Intraspinal infections in patients treated with epidural analgesia]. AB - BACKGROUND: Intraspinal infections (meningitis, epidural abscess) may occur spontaneously or present as a complication of epidural analgesia. MATERIAL AND METHODS: All cases during the 1991-99 period of clinically significant intraspinal infections in patients treated with epidural analgesia were analysed from clinical records in our institution. RESULTS: One patient with uncertain, three patients with well documented meningitis, and one patient with epidural abscess were identified. The treatment time varied from 12 to 49 days in patients with documented infections; the number of catheters varied from one to six. Skin bacteria were isolated from one patient (Staphylococcus sp), opportunistic bacteria (Pseudomonas, Enterococcus, Micrococcus sp) were isolated from three others. Two of the patients were at risk because of probable immunosuppression and chronic infections. Diagnosis and surgery of the patient who developed epidural abscess were significantly delayed because of three negative MRs with and without gadolinium enhancement. INTERPRETATION: Because of the danger of infection related to epidural analgesia, all patients have to be properly monitored as long as they have epidural catheters and also after the removal of catheters. Some epidural abscesses spread longitudinally and may present as a diffuse process on MR without mechanical compression of the medulla, and may be interpreted as negative findings. Myelography with CT scan is an alternative method of investigation in such cases. Early neurosurgical diagnosis and intervention may prevent serious complications. PMID- 11475226 TI - [Diagnostic DNA testing for Huntington disease]. AB - Direct DNA testing for the autosomal dominant neurodegenerative disorder, Huntington's disease, may be performed in a diagnostic setting, as a presymptomatic procedure or prenatally. This paper is intended for physicians practising outside departments of medical genetics who are considering diagnostic testing of patients presenting with symptoms or signs compatible with Huntington's disease. It offers a brief overview of practically relevant clinical, epidemiological, molecular and legal aspects of diagnostic genetic testing for Huntington's disease in Norway. We stress the need for adequate information before sampling and after the test has been performed, and for close contact with the genetics centre which offers the test for Huntington's disease and provides genetic counselling. As with other diagnostic tests, the treating physician is responsible for informing the patient about the result of the test and for ensuring adequate follow-up. The physician will often need the assistance of an expert in clinical genetics. A positive DNA test for Huntington's disease in a patient may have a profound impact on family members, who should be offered genetic counselling and support. Since asymptomatic at-risk family members may ask for a presymptomatic test in the future, diagnostic confirmation at the DNA level is warranted in any person examined because of clinical signs of Huntington's disease, even when the clinical diagnosis is considered unquestionable. PMID- 11475227 TI - [Genetic counseling in presymptomatic testing for Huntington disease]. AB - Norwegian law and international guidelines require genetic counselling before, during and after presymptomatic testing for Huntington's disease. The genetic counselling of at-risk persons who considers taking tests, includes explanation of the possible implications of a test result for both participant and relatives. The test is performed only when explicitly requested by the participant and after informed consent. The participant decides if and when the test should be conducted. The participant also has major influence on the timing of the consecutive phases of the testing procedure, in compliance with medical and ethical recommendations. This paper reviews main issues raised during genetic counselling and the preparation period preceding the test and communication of the test result. We illustrate different individual situations and backgrounds for considering presymptomatic testing for Huntington's disease by describing three anonymized cases and associated pedigrees. PMID- 11475228 TI - [The Norwegian procedure in connection with presymptomatic testing for Huntington disease]. PMID- 11475229 TI - [Need of better knowledge of genetic tests among Norwegian physicians]. AB - BACKGROUND: New predictive genetic tests are introduced in clinical work, and this means new tasks for the physician. MATERIAL AND METHODS: Every sixth member of the Norwegian Medical Association practising as a general practitioner, neurologist or psychiatrist (N = 732), were asked to answer a mailed, anonymous questionnaire about their attitudes to the new tasks. RESULTS: We obtained 451 (62%) answers. There were no significant differences between the various groups in the profession. So far, 54% had no patient in their practice who had taken a predictive genetic test. About two-thirds answered that the geneticist should inform about what is known about the consequences after a test result has been given. The general practitioner wants to do the follow-up. 97% of physicians think that the test result could lead to increased distress in various ways for the tested person or his/her family. Half of the physicians would advise taking a prenatal test if one of the parents had a known risk of an inherited disease and the foetus was at risk. Only 22% are in favour of abortion if the foetus has the gene in question. INTERPRETATION: The physicians (93%) do not think they have sufficient knowledge about predictive genetic tests to handle the information procedure on their own. They want courses in medical genetics, concise and relevant information from geneticists, and the possibility of consulting with specialists. PMID- 11475230 TI - [Can postoperative nausea or vomiting be predicted?]. AB - BACKGROUND: In order to identify patients who may benefit from prophylactic anti emetic medication during surgery, it is of interest to evaluate risk factors for postoperative nausea and vomiting preoperatively by a simple scoring system. MATERIAL AND METHODS: In this retrospective study we evaluated the ability to assess the risk of PONV by a preoperative clinical score developed in our institution (U-score, 0-13 points) and two simple scoring systems from the literature (A-score, 0-4 points and K-score, 0-5 points). 120 patients were interviewed 24 hours after a surgical procedure. Possible risk factors for PONV were scored and compared to the occurrence of PONV. RESULTS: 60% of patients experienced PONV during the first 24 postoperative hours. No difference was seen between those who had received droperidol prophylaxis and those who had not. A significant correlation (p < 0.01) were seen on all scoring systems between occurrence of PONV and high score. With the U-score, 78% of the patients with a score > 7 experienced PONV, compared to only 41% of those with a score < 6. INTERPRETATION: We suggest that patients with a U-score > 7 should receive effective prophylaxis against PONV, i.e. not droperidol only. Further evaluation of different scoring systems should be carried out in larger patient populations. Simplified systems, such as the A-score or K-score or simplified U-score, may be sufficient in uniform surgical populations. The U-score may be more universal because anaesthetic method and use of postoperative opioids are also included. PMID- 11475231 TI - [Plasma concentration analysis of drugs by immunologic techniques]. PMID- 11475232 TI - [Can type 2 diabetes be prevented?]. AB - BACKGROUND: The prevalence of type 2 diabetes among Norwegian men increased from 2.6% to 3.3% from 1986 to 1997. The most important environmental risk factors for type 2 diabetes are obesity and reduced physical activity. Genetic factors are also strongly involved. Biochemical risk factors are impaired glucose tolerance and decreased insulin response. MATERIAL AND METHODS: Only a few small studies have investigated the possibility of primary prevention of type 2 diabetes. RESULTS: In a six-year intervention study on persons with impaired glucose tolerance in China, diet and/or increased physical activity reduced the risk of type 2 diabetes by 30 to 50%. Similar results were found in a study from Sweden. No drug is shown to prevent type 2 diabetes. Possible candidates are metformin and thiazolidinediones which increase insulin sensitivity, and pancreatic lipase inhibitors which reduce the absorption of fat from the gut. Three large, randomised, prospective studies are investigating whether life style intervention or medication can prevent the disease. The results of these studies will be available in about five years. INTERPRETATION: Present evidence clearly indicates that increased physical activity and diet can prevent the development of type 2 diabetes. PMID- 11475233 TI - [Hypoglycemic treatment of type 2 diabetes]. AB - BACKGROUND: Hyperglycaemia in type 2 diabetes results from reduced beta-cell function and insulin resistance. The treatment of type 2 diabetes must be targeted against both conditions to reduce symptoms of hyperglycaemia and the risk of diabetic late complications. MATERIAL AND METHODS: In this review we cover the recent scientific documentation from long term studies on treatment effects. We also discuss the different treatments available. RESULTS: Recent clinical trials have shown that lowering blood glucose levels over time significantly reduces development of microvascular complications in type 2 diabetes. There are several treatment options available which make adequate blood glucose control possible in patients with type 2 diabetes. INTERPRETATION: A reduction in the development of microvascular complications in type 2 diabetes is an achievable goal. PMID- 11475234 TI - [Hypoglycemia--a dreaded complication of diabetes]. AB - Hypoglycaemia is a common side effect of insulin therapy in type 1 diabetes. In patients with type 2 diabetes treated with blood-glucose lowering agents of the sulphonylurea group, hypoglycaemia is less frequent than in insulin-treated patients. In most patients strict metabolic control increases the risk of hypoglycaemia, but this risk may be reduced if patients are offered individualised insulin treatment in combination with active support and education. Previously experienced hypoglycaemic episodes and lack of endogenous insulin production are risk factors for repeated episodes. Patients with longstanding diabetes and loss of warning symptoms have increased risk of severe hypoglycaemic episodes, which may lead to loss of consciousness or convulsions. Driving performance is significantly disrupted at relatively mild hypoglycaemia, and persons with diabetes should not start driving when their blood glucose is in the 4-5 mmol/l range without prophylactic treatment. They ought to have carbohydrate-rich snacks easily available in the car and should stop driving if they feel hypoglycaemic. Repeated episodes of severe hypoglycaemia seem to be associated with cognitive dysfunction. When deciding the targets of blood-glucose lowering therapy, the risk of severe hypoglycaemia must be weighed against the beneficial effects of good metabolic control. PMID- 11475235 TI - [License for elderly physicians]. PMID- 11475236 TI - [Treatment of venous thromboembolism in pregnancy with low molecular weight heparin]. PMID- 11475237 TI - [Acute otitis and antibiotic therapy]. PMID- 11475238 TI - [When money is short, research is set aside. And when the drug industry helps, its interests dominate]. PMID- 11475239 TI - [Pharmacology at the start of a new millennium. Larger assortment and individual drug dosing are coming]. PMID- 11475240 TI - [More stringent requirements in connection with the choice of drugs. Members of the drug committees in Stockholm deliver an annual declaration of challengeability]. PMID- 11475241 TI - [The elderly yesterday, today and tomorrow. Aspects of cohort differences and prevention of the manifestations of aging]. AB - Larger and different groups of elderly persons will meet us in the next few decades. This review describes cohort differences during a couple of decades from the gerontological and geriatric population studies in Goteborg, Sweden (H 70), regarding for example anthropometry, hearing, cognitive function, dietary habits and oral health. The sometimes great cohort differences have an obvious impact on for example planning of health care. Preventive possibilities are dealt with within the fields of nutrition and physical activity. PMID- 11475242 TI - [Epidemiology of aging with focus on physical and mental functional ability]. AB - In the past decades, the "graying" of the population has emerged as a world-wide phenomenon, leading to an increased interest in research on aging. Many population-based studies have been initiated in several countries, such as the Kungsholmen Project in Stockholm, Sweden. These studies have shown that older adults can be recruited to participate in intensive physiological and clinical evaluations, and that longitudinal surveys are well accepted by the elderly. Comorbidity and physical and mental functioning have emerged as important variables for describing health status and identifying risk factors. Dementia arose as one of the most common diseases in the very old, as dementia prevalence nearly doubles every fifth year. Some risk factors for Alzheimer's disease have been identified and interesting working hypotheses have been suggested. The natural history of the dementias have been sufficiently outlined for allocating medical and social resources, and for counseling patients and relatives. PMID- 11475243 TI - [Restructuring of health care services--mostly consequences for the elderly]. PMID- 11475244 TI - [Leflunomide--the first specific disease-modifying drug against rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a disabling disorder with chronic inflammation leading to significant disability and pain if not treated early and effectively. Leflunomide is the first drug developed and registered specifically for modifying disease course in RA, and is a good complement to other forms of drug therapy currently in use. In clinical trials, leflunomide has been proven to exert an effect on clinical parameters of RA-inflammation superior to placebo and equal to methotrexate and sulphasalazine, while clinical response evidently begins somewhat earlier than on these other drugs. There appears to be a retarding effect on radiographic progression over 13 months comparable to that of methotrexate and sulphasalazine. The very long elimination half-life as well as side effects including nausea, diarrhea, and elevated liver enzymes (almost as frequent as with methotrexate) hamper its usefulness, and its definitive place in modern RA-therapy remains to be established. PMID- 11475245 TI - [A case report. Symptoms of vitamin B12 deficiency are not always according to the textbook]. PMID- 11475247 TI - [Assisted euthanasia]. PMID- 11475246 TI - [Treatment with beta-interferon should be started already after the first attack of MS]. PMID- 11475248 TI - [Honey jars and diagnosis of urinary tract infections--ascent quality work]. PMID- 11475249 TI - [Searching for a scope--for the rest of us]. PMID- 11475250 TI - [An officer and a head of a clinical department]. PMID- 11475251 TI - [Gene therapy and cell therapy can be revolutionary for cardiology. An alternative in threatening heart failure after myocardial infarction]. PMID- 11475252 TI - [Gender perspective is neglected in clinical research--mostly for women's disadvantage]. PMID- 11475253 TI - [Adults with congenital heart defects--a growing patient group]. AB - The number of grown ups with congenital heart disease (GUCH) increases due to the success story of pediatric cardiology and heart surgery. However, late complications such as arrhythmia, endocarditis and heart failure are common, as are patients requiring reoperation/catheter intervention. In some categories of congenital heart disease, pregnancy may pose a substantial threat. Early information about this is essential. The care of these patients, including heart surgery, should be centralized. A registry of GUCH-patients has been created in Sweden, promoting experience and knowledge concerning a "new" and expanding group of patients in cardiology. PMID- 11475254 TI - [Organ transplantation, ethics and culture in Japan. Japanese citizens may choose between cardiac death and brain death]. AB - The first heart transplantation in Japan was performed in 1968. However, the surgeon was prosecuted, and transplantations from cadaveric donors came to an end. A new law in 1997 offered the possibility to choose how one would be pronounced dead, and if one chose to be declared dead according to brain related criteria, then one could become a cadaveric donor, if certain conditions were fulfilled. The first donations and transplantations took place in 1999. The Japanese public has been reluctant to accept the concept of brain death. The integrity of the dead body seems to command greater respect in Japan than in Sweden, and there is a reluctance to treat body parts as exchangeable. At the same time many Japanese travel abroad to obtain organs. The act of 1997 is a very interesting way of trying to integrate a new technology in Japanese culture, and the process of obtaining informed consent from relatives relies on traditional cultural roles. PMID- 11475255 TI - [Irritable bowel syndrome. Survey of definitions, differential diagnosis and pathogenesis]. AB - Abdominal pain/discomfort, bloating, need to rush to the toilet, straining, feeling of incomplete bowel emptying and alternating periods of diarrhea and constipation is the clinical definition of the irritable bowel syndrome. The internationally used syndrome definition is based on expert opinions and answers to patient questionnaires. When symptoms are registered prospectively, abdominal pain starts or worsens after meals and is not relieved by defecation. As in the general population patients with the syndrome define diarrhea as loose stools and constipation as hard stools regardless of stool frequencies. Variation in defecatory symptoms and discrepancies between these symptoms and stool consistency are the hallmarks of the syndrome, and the degree of variation per fortnight is relatively stable in the individual patient. Fermentation of carbohydrates by colonic bacteria, increased sensitivity to bowel distention by gas, gas-producing food, increased secretion of cholecystokinin after fatty meals and/or increased sympathetic nerve tone at stress can give rise to symptoms. Symptoms can start after a single period of bacterial gastroenteritis. Although patients seeking medical care for the syndrome are more often anxious, the syndrome itself is not psychosomatic. Symptoms are possibly mediated through partial degranulation of mast cells in bowel mucosa, but this does not make it an allergic disease. If bowel dysmotility can be measured, early stage or a mild case of intestinal pseudoobstruction should be considered. Hyperreactivity in the enteric nervous system and/or in the brain is the likely main cause of the symptoms. More widespread activity in the brain after exposure to stimuli originating from bowel nerves or less inhibition of this stimulation in the brain are possible mechanisms. PMID- 11475256 TI - [Functional gastrointestinal diseases. More reliable diagnosis using new criteria. Modification required in primary care, however]. AB - The diagnosis of functional gastrointestinal disorders is symptom based. New viewpoints go beyond the old concept of a disease-based reductionistic model, according to which effort is directed toward identifying a single underlying biological etiology, to a more integrated biopsychosocial model of illness. "Rome II: A Multinational Consensus Document on Functional Gastrointestinal Disorders" is an attempt to condense present knowledge into a practical systematic manual. Whereas the ordinary clinician may benefit from it in the understanding of the often confusing accumulation of alternating bowel complaints, its main area of application will be international studies. It will also serve as a basis for taking the concept of functional gastrointestinal disorders further. PMID- 11475257 TI - [CDT a valuable marker of overconsumption of alcohol. Guidelines for its use in connection with automobile driver examination]. AB - According to the medical regulations for obtaining a driver's license in Sweden, alcohol abuse/dependency constitutes sufficient grounds for denial. In the case of a conviction for gross drunk driving, it is incumbent upon the offender to present a medical certificate verifying a "sober lifestyle". Biological markers are important tools for proving alcohol abuse in each of these contexts. In this connection, CDT analyses play a key role through their high marked specificity for increased alcohol consumption. The authors have agreed upon the guidelines as presented in this paper for determining sobriety as it pertains to possession of a driver's license. Special emphasis is placed on how CDT tests should be used and interpreted in such contexts, as well as their value as evidence in the case of increased CDT levels. PMID- 11475258 TI - [Reumacon--licensed drug of widespread use]. PMID- 11475259 TI - [Diagnoses of ADHD and DAMP--politics or science?]. PMID- 11475260 TI - [On the epidemiologic evidence of burnout]. PMID- 11475261 TI - [Can radiation hazards of computed tomography be ignored?]. PMID- 11475262 TI - [Vulvar vestibulitis is not necessarily confusing]. PMID- 11475263 TI - [Comments to a proposal: "Specialists closely connected to primary health care" should be excellent generalists in the long run]. PMID- 11475264 TI - [Scientific position doesn't mean immunity against criticism]. PMID- 11475265 TI - [Risk of confusing the vaccines]. PMID- 11475266 TI - [Orthopedics for general practitioners--valuable education]. PMID- 11475267 TI - New products and new technologies for diabetes. PMID- 11475268 TI - Optical radiation safety considerations for ocular glucose monitoring. AB - The potential for noninvasive detection of blood glucose is an area of intense academic and commercial research and a subject of keen interest in the diabetic and healthcare communities. A number of techniques are under investigation that attempt to infer blood glucose levels from measurements of optical signals. Frequently, these techniques are based on laser sources that may, under certain circumstances, be capable of inducing ocular injury. This article provides an overview of ocular damage mechanisms and the international standards for laser exposure limits that have been developed. The application of relevant standards to specific implementations of lasers in optical glucose sensing is presented. In addition, the concept of risk versus benefit for consideration of new medical devices is also discussed. PMID- 11475269 TI - Pharmacokinetics, pharmacodynamics, and dose-response relationship of repaglinide in type 2 diabetes. AB - BACKGROUND: The pharmacodynamics and dose-response relationship of repaglinide, a novel oral hypoglycemic agent, were evaluated in steady-state treatment of patients with type 2 diabetes. METHODS: Efficacy of repaglinide (0.25 mg, 0.5 mg, 1 mg, 2 mg, and 4 mg) was compared to that of placebo in a double-blind, randomized, parallel-group, 4-week dose-response clinical trial in 143 patients. Repaglinide was administered 15 minutes before meals (breakfast, lunch, and dinner). Efficacy of repaglinide therapy was assessed by measuring changes from baseline in mean levels of blood glucose (BGmean), fasting serum glucose (FSG), and mean levels of serum insulin (INSmean). RESULTS: Blood concentrations of repaglinide were proportional to the dose administered. INSmean values increased in all repaglinide treatment groups (by 6.7 to 12.9 microU/mL). All doses of repaglinide significantly decreased values of BGmean and FSG as compared with the placebo group. BGmean values stabilized between the second and third week of repaglinide treatment. A well-defined dose-response relationship was observed for BGmean and FSG values. All doses of repaglinide were well tolerated, and there were no serious adverse events. CONCLUSIONS: These findings show that the therapeutic reduction of serum glucose levels produced by repaglinide is dose dependent for the 0.25- to 4-mg dose range. All doses of repaglinide tested were effective and well tolerated in patients with type 2 diabetes. PMID- 11475270 TI - Repaglinide dose response? A clinician's viewpoint. PMID- 11475271 TI - The importance of rapid insulin secretion: revisited. PMID- 11475272 TI - Continuous glucose monitoring: long-term implantable sensor approach. AB - Problems with existing glucose monitoring technology have resulted in poor compliance with recommended monitoring guidelines by patients with diabetes. To achieve the goal of tight glucose control by patients with diabetes, a long-term implantable glucose sensor should meet the following functional requirements: it should be a one-time minimally invasive implantable with a wireless external unit; provide on-demand real-time glucose levels and trends; operate for up to 12 months after implantation with infrequent recalibration; contain built-in hypoglycemic and hyperglycemic alarms; and have an ergonomically designed, external, wearable user interface. Measurements of glucose in interstitial fluid (ISF) can be used for long-term monitoring. A novel approach to continuous and long-term glucose sensing could be based on measuring the changes in fluorescence of glucose-sensitive indicator molecules. To measure these changes in fluorescence a miniature optoelectronic device with a glucose sensitive indicator could be implanted subcutaneously for long-term remote operation. The fluorescence-based glucose sensing process is reversible and does not consume glucose. The combination of fluorescence-based glucose detection, sensor miniaturization and the use of biomaterials, inducing neovascularization at the implant site, opens the opportunity for achieving the requirements for long-term, continuous and convenient glucose monitoring. PMID- 11475273 TI - Current views on the mechanism of action of thiazolidinedione insulin sensitizers. AB - Resistance to the action of insulin in its target tissues in a major predisposing factor for the development of type 2 diabetes and is also tightly associated with a common pattern of cardiovascular risk factors that characterize the "insulin resistance syndrome." The thiazolidinediones are a new class of drugs that act as insulin sensitizers with well-documented-efficacy in the control of hyperglycemia in patients with overt diabetes. A growing body of evidence also suggests that thiazolidinediones may preserve beta-cell function and protect cardiovascular and renal function in patients with type 2 diabetes. This review will summarize our current notions of the mechanism of action of thiazolidinediones, which appears to involve a fascinating interplay between the partitioning of triglyceride stores, circulating free fatty acids and insulin signaling pathways. A detailed understanding of the action of thiazolidinediones will provide new insights into the pathogenesis of insulin resistance, diabetes and some of the causes of increased cardiovascular mortality in these conditions. PMID- 11475274 TI - Obesity medications and the treatment of type 2 diabetes. AB - Obesity is a chronic disease and requires ongoing treatment. Type 2 diabetes is associated with obesity and improves with weight loss. Diets of 800 kcal/d induce twice the weight loss induced by weight loss medications. The strength of weight loss medication, which should be used with diet and a lifestyle change program, is the maintenance of weight loss. Sibutramine and orlistat are the only two medications approved for the long-term treatment of obesity. Orlistat gives a reduction of low-density lipoprotein (LDL) cholesterol in excess of that expected with weight loss, and the drop in blood pressure expected with weight loss is not seen with sibutramine. Except in newly diagnosed patients with diabetes subjects, patients with diabetes lose half the weight of subjects who do not have diabetes when treated with weight loss medications. Metformin and, to a lesser extent, acarbose cause weight loss, making them attractive choices for the treatment of obese type 2 diabetic subjects. Repaglinide appears to be weight-neutral, but other medications for patients with diabetes can be associated with weight gain. Many new medications are in development for the treatment of obesity. These new medications act through a variety of mechanisms and will surely play an increasingly important role in the treatment of obese patients with type 2 diabetes. PMID- 11475275 TI - Assessing body composition of adults with diabetes. AB - Overweight and obesity are associated with the development of type 2 diabetes. Thus, it is important for clinicians to accurately measure and monitor the body composition of at-risk individuals and patients with diabetes. This article reviews valid and reliable field methods and prediction equations for assessing the body composition of obese individuals, as well as persons with type 2 diabetes. We also reviewed research that assessed the validity of practical methods in estimating the body composition of individuals with either type 1 or type 2 diabetes. PMID- 11475276 TI - Control of glucose homeostasis by incretin hormones. AB - In humans as well as in other animal species, the ingestion of food provides the fundamental source of energy for various cellular activities. The intake of food and the ability of controlling the plasma levels of substrates for energy production involve complex mechanisms that ensure a constantly adequate supply of metabolites both in the fasting and in the fed state. A number of hormonal peptides released from the gastrointestinal (GI) tract in response to the ingestion of food have been shown to play a critical role in the postprandial control of glucose homeostasis. They are known to act through three main mechanisms of action. These include; (1) stimulation of insulin secretion of pancreatic islet (beta) cells; (2) inhibition of hepatic gluconeogenesis by suppression of glucagon secretion; and (3) inhibition of GI motility. While for some of these hormones all three mechanisms of action are utilized under physiological conditions, others preferentially use one or a combination of two mechanisms for lowering postprandial hyperglycemia. Although the term glucoincretins (or incretins, or insulinotropic hormones) etymologically only describes factors capable of inducing insulin secretion, it is more frequently used to identify a larger class of peptides that, rather than manifesting a specific mechanism of action (i.e., insulin secretion), share the ability of controlling glucose excursion in the fed state (with or without a direct insulinotropic effect). The latter more inclusive meaning, incretins, is used in this article. This review summarizes recent advances on synthesis, secretion, blood plasma patterns, and metabolism of some of the major GI regulatory peptides acting in the postprandial state. PMID- 11475277 TI - Inhaled insulin. AB - Inhalation is a potentially viable route of insulin administration. This treatment is being tested but has not been approved by the United States Food and Drug Administration (FDA). The lung airways contain bronchial tubes, which are impermeable to insulin and alveoli, from which insulin can be absorbed into the circulation. Inhaled asthma medications deposit before reaching the alveoli. Novel devices can deliver insulin particles via slow even breaths into the alveoli. An aerosol of either powdered or solubilized insulin can be released mechanically or electronically. Pulmonary toxicity due to inhaled insulin has not been reported, but no chronic use studies have been conducted. The efficiency of insulin delivery by inhalation may be compromised by: (1) losses within the device and the environment; (2) deposition onto the throat and bronchial tubes; and (3) incomplete absorption from the alveoli. With current delivery technology, the maximum possible delivery efficiency is approximately 30%. The peak activity of inhaled regular insulin occurs at 60 minutes, which is similar to that of subcutaneous lispro insulin. Inhaled long-acting insulin is in early development. Studies comparing sequentially administered subcutaneous and inhaled regular insulins have demonstrated significantly reproducible glucose-lowering effects of inhaled insulin. In type 1 and insulin-treated type 2 patients, substitution of premeal inhaled insulin for subcutaneous insulin has resulted in no significant change in control. In a series of patients with type 2 diabetes failing oral agents, addition of premeal inhaled insulin resulted in significantly improved control. Inhaled insulin will become established if ongoing studies continue to demonstrate this technology to be safe and effective. PMID- 11475278 TI - Hopes, dreams, and reality: the long road to pain-free glucose testing. PMID- 11475279 TI - FDA approves new Glucose Monitoring System. PMID- 11475280 TI - Diabetes Prevention Program meets recruitment goals. PMID- 11475281 TI - Database programs for use with blood glucose meters. PMID- 11475282 TI - Noninvasive laser measurement of blood glucose in the eye: a bright idea or an optical illusion? PMID- 11475283 TI - Introduction to concepts in laser technology for glucose monitoring. AB - Lasers are important tools in many diabetes-related applications, both diagnostic and therapeutic. Despite its wide range of uses, the laser derives all of its advantages over more conventional light sources from a few basic principles. A brief introduction to the fundamental properties of lasers emphasizes these points and suggests ways in which they can be exploited in experiments. Short descriptions of particular laser-based methods for monitoring glucose and related molecules provide introductions to research articles on these subjects. PMID- 11475284 TI - Laser-based measurement of glucose in the ocular aqueous humor: an efficacious portal for determination of serum glucose levels. AB - The potential for the noninvasive measurement of blood glucose concentration to improve the level of blood sugar control and the quality of life of diabetic patients has been recognized for nearly 3 decades. Because of the many challenges involved in directly sensing glucose in the bloodstream noninvasively, the approach of noninvasive measurement of glucose concentration in the ocular aqueous humor, which reflects blood glucose concentration, has been likewise pursued over those 3 decades. Moreover, studies of the dynamic relationship between blood glucose concentration and ocular glucose concentration in both animals and humans suggest minimal latency between variations in blood glucose and those in ocular glucose. In a review of technological approaches to such measurements, the most promising techniques appear to be laser-based, and include laser polarimetry and Raman spectroscopy. Results for techniques such as multiwavelength polarimetry and Raman spectroscopy have been quite good, and it is likely that one or both of these techniques, possibly combined with refractive index measurement, will yield a useful instrument for both clinical use and patient self-monitoring. PMID- 11475285 TI - The use of polarized laser light through the eye for noninvasive glucose monitoring. AB - BACKGROUND: In order to optimally manage diabetes mellitus, it is recommended blood glucose levels be monitored several times daily so an appropriate action can be taken to maintain tight control of these levels within a normal physiological range. All commercially available devices to measure blood glucose concentrations require the extraction of a drop of blood, normally obtained via the lancing of a finger. The main drawback to this method is the pain, often leading to low patient compliance. Therefore, a noninvasive glucose sensing method would greatly facilitate the management of diabetes. METHODS: In this article, we describe in vitro and in vivo results from a laser-based optical polarimetry system using the anterior chamber of the eye as a potential method to noninvasively monitor glucose levels in the body. RESULTS: It is shown, in vitro, that glucose can be predicted in the presence of albumin at physiological levels and, through the use of a novel light coupling mechanism, it is demonstrated that a polarimetric signal can be detected, in vivo, through a rabbit eye. CONCLUSIONS: Although the commercial production of a feasible noninvasive glucose monitoring method is still years away, laser-based polarimetry remains a viable alternative due to its potential to extract concentration information using the eye as a unique optical window into the body. PMID- 11475286 TI - A noninvasive glucose monitor: preliminary results in rabbits. AB - BACKGROUND: Glucose in the aqueous humor appears to correlate with plasma glucose in humans. It is therefore a potential substrate for noninvasive optical glucose monitoring techniques. We wished to determine the potential for using rabbit aqueous humor as a model to develop a noninvasive method of measuring glucose with Raman spectroscopy. METHODS: Aqueous humor from 32 rabbit eyes was removed immediately after sacrifice by rapid exsanguination under anesthesia. Raman spectroscopy was performed on the aqueous humor using near infrared (NIR) excitation wavelengths. The Raman spectra from 785 nm and 787.2 nm were subtracted one from the other to eliminate broadband fluorescence. The spectra were then analyzed with linear and nonlinear multivariate analysis and assessed for ability to predict actual aqueous humor glucose concentration. Nine other rabbits were anesthetized with xylazine in order to cause elevation of blood glucose by blocking release of insulin. Blood and aqueous humor glucose were measured at various times after injection of xylazine. Correlation of aqueous humor glucose with simultaneous plasma glucose was assessed. RESULTS: Partial least squares analysis of raw Raman spectra of aqueous humor showed fair correlation with actual glucose concentration (r2 = 0.76). When the fluorescence spectrum was subtracted prior to linear multivariate analysis correlation was good (r2 = 0.90). When back-propagation with an artificial neural network was added to the analysis, correlation was further improved (r2 = 0.98). Aqueous humor glucose concentration exceeded blood glucose concentration at normoglycemic levels. When blood glucose rose above 200 mg/dL, aqueous humor glucose correlated linearly with plasma glucose. CONCLUSIONS: Raman spectroscopy can accurately predict glucose concentration in rabbit aqueous humor in vitro. Although rabbit aqueous humor is probably not a good model of human aqueous humor glucose physiology, its reliable correlation with plasma glucose makes it a good model on which to test noninvasive optical sensing techniques. PMID- 11475287 TI - Reagentless blood analysis by near-infrared Raman spectroscopy. AB - BACKGROUND: Raman spectroscopy has advantages over infrared absorption spectroscopy. Combined with a novel multivariate technique, hybrid linear analysis (HLA), low prediction error is expected. METHODS: A near-infrared (NIR) light source excited Raman signals, and a charge coupled device (CCD) camera was used to collect the signal. Samples were collected from 69 individuals for 7 weeks. The standard multivariate calibration technique, partial least squares (PLS) and HLA were both used to analyze the collected spectra. A Clarke error grid was used to evaluate the usefulness of the glucose measurement in serum. RESULTS: The root mean squared error of prediction (RMSEP) for glucose in serum obtained with PLS is 21 mg/dL, and the RMSEP obtained with HLA is 17 mg/dL. In whole blood, the PLS RMSEP for glucose was 79 mg/dL, and HLA predictions had an RMSEP of 63 mg/dL. CONCLUSIONS: The measurement technique was robust over the 7 week period. HLA was shown to generate a lower prediction error than PLS. The predictions by both PLS and HLA were clinically acceptable. The result with whole blood requires further improvement. PMID- 11475288 TI - Use of dynamic light scattering and Scheimpflug imaging for the early detection of cataracts. AB - Cataract is a leading cause of blindness. Diabetes and glycemic conditions enhance the chances of developing cataracts early. At clinical stage cataracts are detected and documented by slit-lamp biomicroscopy and Scheimpflug photography. A compact fiber optic probe, developed for space experiments, was mounted on a Scheimpflug imaging system. The probe detects and documents cataracts at pre-clinical stage noninvasively and quantitatively. The early detection at the molecular level may lead to medical treatment of cataracts, better control of glycemia, and diabetes mellitus. PMID- 11475289 TI - Dynamic light scattering of diabetic vitreopathy. AB - BACKGROUND: Diabetes induces pathology throughout the body via nonenzymatic glycation of proteins. Vitreous, which is replete with type II collagen, undergoes significant changes in diabetes. The resultant diabetic vitreopathy plays an important role in diabetic retinopathy. Detecting these molecular changes could provide insight into diabetic eye disease as well as molecular effects elsewhere in the body. METHODS: Human eyes were obtained at autopsy and studied in the fresh, unfixed state. Sclera, choroid, and retina were dissected off the vitreous for dark-field slit microscopy and dynamic light scattering (DLS). For the former, the entire vitreous was exposed. For the latter, only a window at the equator was dissected in some specimens, and the anterior segment was removed leaving the posterior lens capsule intact in others. DLS was performed to determine particle sizes at multiple sites 0.5 mm apart, spanning the globe at the equator (window dissections) and along the antero-posterior axis. RESULTS: Dark-field slit microscopy in diabetic subjects detected findings typical of age-related vitreous degeneration, but at much younger ages than nondiabetic controls. Noninvasive DLS measurements found a greater heterogeneity and larger particle sizes in vitreous of subjects with diabetes as compared to age-matched controls. CONCLUSIONS: DLS can detect and quantify the early molecular effects that cause vitreous collagen fibrils to cross-link and aggregate. This could provide valuable insight into ocular and systemic effects of hyperglycemia, because the molecular changes in diabetic vitreopathy could serve as an index of such effects throughout the body. In addition to the diagnostic implications, this methodology could provide a rapid, reproducible way to monitor the response to therapy with novel agents intended to prevent the complications of diabetes on a molecular level. PMID- 11475290 TI - Lasers and diabetic retinopathy: the art of gentle destruction. AB - Laser retinal photocoagulation represents the primary therapy for the potentially blinding manifestations of diabetic retinopathy. Advances in laser therapy for diabetic eye disease have arisen from the convergence of laser technology and clinical ophthalmology. In this review, the basis of laser therapy in diabetic retinopathy is discussed. Included are the clinical characteristics, classification, and pathogenetic mechanisms of diabetic retinopathy. The principles of laser technology, variety of lasers currently available for therapy, and mechanisms of laser actions on retinal target tissues are introduced. The milestone clinical trials establishing the efficacy of laser therapy and the parameters for its clinical application in diabetic retinopathy are outlined. PMID- 11475291 TI - Using lasers in diabetic wound healing. AB - Lasers are a treatment choice that appeals to patients. Early research suggests that laser therapy may have a role in hastening wound healing. Attempts have been made to use helium neon, CO2, and KTP lasers in encouraging wound healing in diabetics. We will review the English literature related to laser use in diabetic wound healing and discuss the concept of the use of nondestructive lasers for biostimulation. Further research is needed to assess effectiveness of biostimulation for diabetic wound healing. PMID- 11475292 TI - The best defense against hypoglycemia is to recognize it: is caffeine useful? AB - Caffeine, 1,3,7trimethylxanthine, is used by 80% of the adult population of the world in its various forms. Even the simple pleasure of consuming this socially acceptable drug has implications for the person with diabetes mellitus. Caffeine may increase an individual's sensitivity to hypoglycemia through the combined effects of reducing substrate delivery to the brain via constriction of the cerebral arteries, whilst simultaneously increasing brain glucose metabolism and augmenting catecholamine production. This article summarizes the evidence supporting the hypothesis that caffeine influences the perception of and physiological response to hypoglycemia. Under laboratory conditions, acute ingestion of caffeine markedly enhances the symptomatic and sympathoadrenal responses to hypoglycemia in both healthy volunteers and patients with type 1 diabetes. Recently a study of free-living people with type 1 diabetes showed that caffeine consumption increased the awareness of hypoglycemia. Caffeine has been associated with a number of negative effects and addiction. Most serious of these associations are ischemic heart disease and hypertension, the relationships have not been clearly established and the evidence to date is controversial. Thus we conclude that in modest doses, caffeine may be a useful adjuvant therapy for patients with hypoglycemia unawareness. For once here is a therapy which is inexpensive, safe, and remarkably popular with its consumers. PMID- 11475293 TI - FDA Clinical Chemistry and Clinical Toxicology Devices Panel. PMID- 11475294 TI - The FDA panel advises approval of the first continuous glucose sensor. PMID- 11475295 TI - New technologies and genetics of type 1 diabetes. PMID- 11475296 TI - The Internet may provide a long-awaited answer to health informatics. PMID- 11475297 TI - Database software for use with blood glucose meters. PMID- 11475298 TI - Diabetes information technology & WebWatch. General resources. PMID- 11475299 TI - Radioimmunoassay for glutamic acid decarboxylase-65. AB - BACKGROUND: Glutamic acid decarboxylase-65 (GAD65), the enzyme that catalyzes the formation of gamma-aminobutyric acid (GABA), is the major autoantigen in both type 1 (insulin-dependent) diabetes and stiff-man syndrome (SMS). The observation that GAD65 autoantibodies may be present for years before the clinical onset of diabetes raises the question of when GAD65 is available to initiate an immune response to allow the formation of autoantibodies. In order to address this question it will be necessary to measure GAD65 in tissue, cells, and plasma. METHODS: A radioimmunoassay (RIA) was developed for GAD65 based on the use of a polyclonal rabbit antiserum directed to the N-terminus of GAD65. RESULTS: Using the GAD65 RIA, we have determined the GAD65 content in a human GAD65 gene transfected cell line and in beta-cell preparations from different species. The assay detects an increase of immunoreactive GAD65 after glucose-stimulation and GAD65 that is discharged from rat beta cells after their exposure to the diabetogenic agent streptozotocin. It also measures good recovery of GAD65 added to human plasma samples. CONCLUSIONS: The GAD65 RIA makes it possible to determine both cellular and extracellular levels of GAD65; this might be useful in investigating the mechanisms leading to the formation of GAD65 autoantibodies in type 1 diabetes and SMS patients. PMID- 11475300 TI - Fluorescein kinetics in interstitial fluid harvested from diabetic skin during fluorescein angiography: implications for glucose monitoring. AB - BACKGROUND: Glucose monitoring based on sampling skin interstitial fluid (ISF) is being developed as an alternative to fingerstick blood glucose monitoring. Time delays between rapidly changing levels of glucose in blood and interstitial fluid have been reported in the literature to be between 5 and 20 minutes. This study investigated the time delay between the injection of a small molecular weight fluorescent tracer into the circulation and interstitial fluid. METHODS: Diabetic subjects undergoing fluorescein angiography were studied. Skin ISF was sampled using a proprietary microporation and harvesting process. ISF was drawn through micropores created in the stratum corneum. After intravenous injection of sodium fluorescein, samples of ISF were drawn from 2 sites for 30 seconds over a period of 20 minutes. Fluorescence levels in ISF were measured with a fluorometer and used to create ISF fluorescein concentration versus time profiles. RESULTS: The ISF fluorescein versus time profiles were characterized by a rapid rise followed by a slow decay. The time to peak of the ISF fluorescein concentration ranged from 2-4 minutes for the patients studied. CONCLUSIONS: Intravenous injection of a bolus of low molecular weight fluorescent tracer was used to estimate the time delay between changing glucose levels in blood and ISF. The results indicate that the ISF sampling technology utilized here is capable of tracking rapidly rising levels of blood glucose. PMID- 11475301 TI - Capillary blood sampling for self-monitoring of blood glucose. AB - New and emerging capillary blood sampling technologies for self-monitoring of blood glucose (SMBG) are reviewed for their impact on factors pertaining to users such as pain, and from the standpoint of skin physiology and technical feasibility. Innovative blood sampling techniques based on lancets for skin penetration on nonfinger (alternate) sites such as the forearm seem to be virtually painless, convenient and cost effective as compared to other methods such as laser-based perforation of fingertips. Alternate site blood sampling with new lancet devices appears not only medically sound but also technically practical and user-friendly. It is anticipated that alternate site blood sampling techniques would improve compliance rate and, consequently, outcome of treatment for patients with diabetes. PMID- 11475302 TI - Issues and implications in the selection of blood glucose monitoring technologies. AB - Over the last 2 decades assay technology originating in the laboratory has been adapted for the special situation of in vitro blood glucose monitoring in the home, at work or play, or at the bedside. The availability of blood glucose monitoring devices has had a significant impact on the treatment of diabetes, especially with respect to involving the patients in their treatment. The unique requirements of this type of testing have led to novel developments in sample acquisition techniques, analyte detection, measurement techniques, and error detection. The performance of these in vitro devices in terms of accuracy and imprecision is largely dependent on factors that contribute to variation in response that are related to testing with blood samples outside of the laboratory. These factors include, for example, variations in environmental conditions, the variability of hematocrit and oxygen concentrations of the blood, and the fact that the blood is used undiluted. Therefore, the technologies used have been selected, developed, optimized, and calibrated to minimize the impact of these factors. The technologies also must be capable of providing accurate, reproducible results over the large range of clinical interest from the hypoglycemic range to glucose concentrations 10 to 15 to 20 times greater. However, when selecting a technology there are invariably some trade-offs to consider. Thus, the products must be optimized to balance performance, reliability, and cost. Examples are discussed. PMID- 11475303 TI - Blood glucose monitoring in the cutaneous microenvironment. PMID- 11475304 TI - Experience with the Internet release of AIDA v4.0- http://www.diabetic.org.uk.aida.htm--an interactive educational diabetes simulator. AB - AIDA v4.0 is a freeware computer program that permits the interactive simulation of plasma insulin and blood glucose profiles for demonstration and teaching purposes. It has been made freely available, without charge, on the World Wide Web as a noncommercial contribution to continuing diabetes education. Since its Internet launch in 1996 over 23,000 people have visited the AIDA Web site (http://www.diabetic.org.uk/aida.htm) and over 7,750 copies of the program have been downloaded gratis. This report overviews the Internet release of AIDA v4.0 and provides examples of the simulator in operation. The concept of a "virtual diabetic patient" is introduced. This provides an electronic representation of a patient with diabetes that can be used for self-learning/teaching/demonstration purposes. PMID- 11475305 TI - Diabetes simulators: ready for prime time? PMID- 11475306 TI - Measuring glycated proteins: clinical and methodological aspects. AB - The recognition that nonenzymatic glycation, wherein free sugar condenses with certain reactive protein amino groups, is increased in hyperglycemic states has led to improved means for assessing diabetic control through the measurement of concentrations of glycated proteins in the circulation. This article reviews the chemistry, methods for measurement, and clinical relevance of circulating glycated proteins in the management of diabetes. PMID- 11475307 TI - Functional genomics and the discovery of new drug targets. AB - Functional genomics can be defined as the search for the physiological role of a gene for which only its primary sequence is known. One example of a successful functional genomics adventure is the search for the natural ligands of orphan G protein-coupled receptors (GPCRs). GPCRs are proteins containing 7 hydrophobic domains that are the recognition sites of neurotransmitters and neuropeptides. Although many of these have been shown to interact with known natural ligands, several bind ligands that have not been thus far isolated. These are the so called "orphan" GPCRs. As an example of functional genomics, an "orphan receptor strategy" has been developed to identify the natural ligands of orphan GPCRs. We describe that the application of this strategy has already led to the identification of 4 new neuropeptides and report on what has been learned about these neuropeptides. We finally discuss the importance of the application of the orphan receptor strategy to the development of novel drugs. PMID- 11475308 TI - Transcutaneous electrostimulation: emerging treatment for diabetic neuropathic pain. AB - Three independent studies utilizing transcutaneous electrical nerve stimulation to relieve diabetic peripheral neuropathic pain were reviewed. The proprietary equipment, an H-wave machine, administered all electrotherapy. The first two studies assessed the efficacy of electrotherapy alone and electrotherapy with amitriptyline. The treated electrotherapy group reported an overall greater reduction of symptoms, 52% with 2-3 weeks of active treatment. Amitriptyline alone produced a 26% reduction of pain after 4 weeks. The addition of active electrotherapy to amitriptyline produced a 66% reduction of pain. The final study looked at patients who have utilized electrotherapy for over one year. A reported 44% improvement of symptoms was attained with continuous electrotherapy treatment. The data also suggested that a maintenance treatment protocol for long term pain relief would have to be developed. PMID- 11475309 TI - Transcutaneous electrical nerve stimulation: new applications of an established treatment. PMID- 11475310 TI - Novel approaches to cellular transplantation from the U.S. Space Program. PMID- 11475311 TI - Programmable implantable insulin infusion devices and diabetes care. PMID- 11475312 TI - Stains in microbiology and molecular biology. PMID- 11475313 TI - Use of the gram stain in microbiology. AB - The Gram stain differentiates bacteria into two fundamental varieties of cells. Bacteria that retain the initial crystal violet stain (purple) are said to be "gram-positive," whereas those that are decolorized and stain red with carbol fuchsin (or safranin) are said to be "gram-negative." This staining response is based on the chemical and structural makeup of the cell walls of both varieties of bacteria. Gram-positives have a thick, relatively impermeable wall that resists decolorization and is composed of peptidoglycan and secondary polymers. Gram-negatives have a thin peptidoglycan layer plus an overlying lipid-protein bilayer known as the outer membrane, which can be disrupted by decolorization. Some bacteria have walls of intermediate structure and, although they are officially classified as gram-positives because of their linage, they stain in a variable manner. One prokaryote domain, the Archaea, have such variability of wall structure that the Gram stain is not a useful differentiating tool. PMID- 11475314 TI - Application of stains in clinical microbiology. AB - Stains have been used for diagnosing infectious diseases since the late 1800s. The Gram stain remains the most commonly used stain because it detects and differentiates a wide range of pathogens. The next most commonly used diagnostic technique is acid-fast staining that is used primarily to detect Mycobacterium tuberculosis and other severe infections. Many infectious agents grow slowly on culture media or may not grow at all; stains may be the only method to detect these organisms in clinical specimens. In the hands of experienced clinical microscopists, stains provide rapid and cost-effective information for preliminary diagnosis of infectious diseases. A review of the most common staining methods used in the clinical microbiology laboratory is presented here. PMID- 11475315 TI - Staining nucleic acids and proteins in electrophoresis gels. AB - This review addresses the most widely used stains for gel electrophoresis in molecular biology today. A brief introduction to electrophoresis is given, followed by a description of agarose and acrylamide gel formation. The major stains for nucleic acids are described, most of which are fluorescent and intercalate into the double helix structure of the nucleic acids. An ultraviolet transilluminator is necessary to visualize the fluorescent bands. The major protein stains include Coomassie blue, silver, and commercial fluorescent stains. Negative staining also can be used to stain the gel background, leaving the protein bands of interest transparent and visible against a white background. PMID- 11475316 TI - Application of the membrane filter technique to bromodeoxyuridine immunochemistry for exfoliative cytology. AB - The membrane filter technique for smear specimens of tumors in bromodeoxyuridine (BrdU) immunochemistry is described. The staining results of Raji cells processed using the filter technique was compared with that obtained by the conventional cytospin method. Although the BrdU mean labeling index (LI) for in cytospin specimens was almost the same as the LI in membrane filter specimens, filter specimens showed excellent staining and less cell destruction compared with those processed by cytospin. Small amounts of tumor specimens such as squamous cell carcinoma and polymorphous low-grade adenocarcinoma also were processed using the membrane filter appliance. For squamous cell carcinoma, the LI for the filter specimens was 5.36+/-0.38 and that of the paraffin sections was 5.56+/-0.38. The membrane filter technique provided relatively undamaged specimens for exfoliative cytology and will be useful for immunohistochemical evaluation of tumor cells and for routine, noninvasive cytological screening. PMID- 11475317 TI - A contribution to the theory of biological staining based on the principles for structural organization of biological macromolecules. AB - Biological staining is to a large degree explainable based on the principles governing folding and aggregation of macromolecules in aqueous solution. Most macromolecules are polyions, which, except for heteropolysaccharides, have a large proportion of nonpolar or only slightly polar residues. Because they are amphiphilic, they react in water by a complex set of hydrophobic interactions involving charged residues, nonpolar residues and water molecules. The hydrophobic interactions lead to complex folding systems or micelle-like structures. Dyes are amphiphilic molecules with a tendency to form micelles, but with limitations due to geometric constraints and charge repulsion. Macromolecules and dyes react with each other in aqueous solution following the same principles as for the structural organization of macromolecules, as in protein folding for example. Dye binding requires near contact between nonpolar groups in both the dye and macromolecule, and this is accomplished by choosing a pH at which the dye and macromolecule have opposite net charges. Charge attraction is insufficient for binding in most cases, but it is directive because it determines which macromolecules a given dye ion is able to contact. These considerations apply to the staining of globular (cytoplasmic) proteins and to nucleic acid staining. The staining mechanism is by hydrophobic interactions. Above approximately pH 3.5, DNA may also bind dyes by hydrophobic intercalation between the bases of the double helix; at lower pH the double helix opens and dye binding is as for RNA and globular proteins. Heteroglycans (mucins) have virtually no nonpolar groups, so nonpolar interactions are restricted to the dye molecules. Metachromatic staining of heteroglycans is due to hydrophobic bonding or micelle formation between the monovalent planar dye molecules aided by charge neutralization by the negatively charged heteroglycans. Alternatively, as the charge attraction increases with the number of closely placed charges, acidic heteroglycans may be stained by a polycation such as alcian blue or colloidal iron. For elastic fiber and collagen staining, actual hydrophobic interactions are less important and hydrogen bonding and simple nonpolar interactions play a major role. These macromolecules may therefore be stained using a nonaqueous alcoholic solution. PMID- 11475318 TI - Physical fitness and changes in mortality: the survival of the fittest. AB - Physical fitness is a term describing maximal aerobic capacity adjusted for body size and composition, and is an integrated measure of cardiorespiratory and neuromusculo-skeletal function, oxygen transport and delivery, and psychological drive. Accordingly, high physical fitness requires that all these important body functions function normally, while low physical fitness suggests malfunction of one or more of them. High levels of physical fitness--maintained through heavy daily exercise--has probably been a necessary requirement for survival in the earlier history of humans. In modern, industrialised countries the demand for physical activity to sustain life is declining, and a decline in physical fitness is observed in many populations. Epidemiological studies over the past 50 years have unanimously shown that low physical activity and physical fitness is associated with high cardiovascular and total mortality. Recent data also suggest that low physical activity and physical fitness is followed by an increased incidence of diabetes mellitus and nonfatal cardiovascular diseases. A number of unfavourable biochemical and physiological aberrations following physical inactivity probably explain these observations. However, recent studies also indicate that changes in physical activity, and especially changes that bring increases in physical fitness, can reverse this rather pessimistic scenario. The Global Burden of Disease Study initiated by the World Health Organization included physical inactivity among the most important risk factors threatening global health. A sedentary life style may be as detrimental to health as smoking. Encouragement of physical activity is an important and difficult task, as society is becoming increasingly successful in reducing our need to move. PMID- 11475319 TI - Zinc status in athletes: relation to diet and exercise. AB - Zinc is involved in the biochemical processes supporting life, such as cellular respiration, DNA reproduction, maintenance of cell membrane integrity and free radical scavenging. Zinc is required for the activity of more than 300 enzymes, covering all 6 classes of enzyme activity. Zinc binding sites in proteins are often of distorted tetrahedral or trigonal bipyramidal geometry, made up of the sulphur of cysteine, the nitrogen of histidine or the oxygen of aspartate and glutamate, or a combination. Zinc in proteins can either participate directly in chemical catalysis or be important for maintaining protein structure and stability. The nutritional habits of elite athletes during training and competition are quite different from the recommended diet in the majority of the population. Endurance athletes often adopt an unusual diet in an attempt to enhance performance: an excessive increase in carbohydrates and low intake of proteins and fat may lead to suboptimal zinc intake in 90% of athletes. Mild zinc deficiency is difficult to detect because of the lack of definitive indicators of zinc status. In athletes, zinc deficiency can lead to anorexia, significant loss in bodyweight, latent fatigue with decreased endurance and a risk of osteoporosis. PMID- 11475320 TI - Plasma leptin and exercise: recent findings. AB - The cloning of murine and human obese genes in 1994, and the subsequent identification that the product of the obese gene, leptin, is secreted from adipose tissue, stimulated a tremendous amount of interdisciplinary interest in adipose tissue endocrinology and the potential role of this tissue in the regulation of energy balance. Exercise, with concomitant changes in fuel flux, systemic hormone levels and energy expenditure, may contribute to the regulation of plasma leptin levels and presumably, leptin action. The initial work characterising the leptin-exercise relationship was equivocal. Cross-sectional studies provided some mixed evidence regarding the relationship between aerobic capacity or habitual physical activity and plasma leptin. In contrast, studies on acute bouts of exercise and exercise training interventions have, with few exceptions, suggested that exercise does not alter systemic leptin independent of changes in fat mass. In general, these studies did not carefully control for energy balance, and sampled only a single fasting plasma leptin level. Two recent studies utilising experimental designs in which energy balance was controlled and 24-hour profiles of plasma leptin were determined have provided the most compelling evidence to date of the interaction between exercise, energy balance and systemic leptin in humans. These studies provide a clear explanation for the apparent lack of an acute effect of exercise on systemic leptin and underscore the importance of clearly defining the balance between energy intake and energy expenditure when studying the physiology of leptin. The aim of this brief review is to provide an overview of the interaction between energy expenditure during physical activity and systemic leptin level. Special emphasis will be placed on those studies in which energy intake/balance was carefully controlled. PMID- 11475321 TI - Exercise and the child born prematurely. AB - With the increase in survival rates of children born prematurely, issues related to their active pursuits and responses to exercise have been gaining increasing attention. In some preterm children with an extremely low birthweight, bronchopulmonary dysplasia or cerebral palsy exercise capacity may be limited, especially in tasks requiring good neuromotor coordination. Deficiencies in aerobic and anaerobic performance, strength and coordination may even occur in children without overt manifestations of a neuromuscular or pulmonary disease. However, as a rule, children born prematurely may engage in physical activities and competitive sports without limitations. Exercise is safe in almost all such children as long as precautions are taken to avoid exercise-induced bronchoconstriction. However, to date there are no studies that have determined the efficacy of training. A wide variety of activities should be encouraged in all children born prematurely at an early age, to support the development of skills and to compensate for the possible effects of their premature birth on coordination. PMID- 11475323 TI - Physical activity guidelines for children and adolescents: a critical review. AB - Since it is generally accepted that the onset of many chronic diseases lies in early childhood, preventive strategies should start as early in life as possible. Physical inactivity is recognised as an important determinant for chronic disease and the prevalence of physical inactivity is increasing during adolescence. Therefore, in the last 10 years much effort has been put into the development of physical activity guidelines for children and adolescents. In this critical review it is shown that the scientific evidence on which these guidelines are based is rather weak. There is only marginal evidence that physical activity during youth is related to adult health status. There is, however, hardly any evidence for a certain dose-response relationship or a particular threshold value from which guidelines can be obtained. Furthermore, it can be suggested that although the value of physical activity guidelines for children and adolescents for public health purposes is beyond doubt, perhaps these guidelines should focus on aspects other than possible health benefits. PMID- 11475322 TI - Prevalence and mechanisms of development of asthma and airway hyperresponsiveness in athletes. AB - A high prevalence of asthma and airway hyperresponsiveness (AHR) has been reported in the athlete population. Factors potentially predisposing athletes to these conditions have not been clearly identified. Although moderate exercise has been shown to be beneficial in patients with asthma, repeated high-intensity exercise could possibly contribute to the development of asthma and AHR. This report provides an overview of the prevalence and possible mechanisms of development of asthma and AHR in the athlete population. The prevalence of asthma and AHR are higher in athletes than in the general population, particularly in swimmers and athletes performing sports in cold air environments. Possible mechanisms involved in the development of asthma in athletes are still uncertain; however, the content and physical characteristics of the inhaled air seem to be important factors, while immune and neurohumoral influences could play a modulatory role. This report stresses the need for further studies to better define the aetiologic factors and mechanisms involved in the development of asthma and AHR in athletes, and proposes relevant preventive and therapeutic measures. PMID- 11475325 TI - Analysis of Tag1-like elements in Arabidopsis thaliana and their distribution in other plants. AB - Analysis of genomic DNA of Arabidopsis Columbia (Col.) ecotype using a transposon Tag1-specific primer showed the presence of Tag1 homologues which was confirmed by Southern hybridization with a Tag1 probe. Further analysis showed that the homologue, 0.75 kb in length, had inverted repeats at both ends, 8-bp duplicated sequences at the site at which it is located and about 80% homology with Tag1, and was randomly distributed in the Arabidopsis genome. Based on these results, we concluded that these elements are non-autonomous variants of Tag1 and we termed this element sTag1. Using the polymerase chain reaction fragment hybridization technique, we found the distribution of such homologues in other plant species. PMID- 11475324 TI - Recommendations for grading of concussion in athletes. AB - Mild sports-related concussions, in which there is no loss of consciousness, account for >75% of all sports-related brain injury. Universal agreement on concussion definition and severity grading does not exist. Grading systems represent expertise of clinicians and researchers yet scientific evidence is lacking. Most used loss of consciousness and post-traumatic amnesia as markers for grading concussion. Although in severe head injury these parameters may have been proven important for prognosis, no study has done the same for sport-related concussion. Post-concussion symptoms are often the main features to help in the diagnosis of concussion in sport. Neuropsychological testing is meant to help physicians and health professionals to have objective indices of some of the neurocognitive symptoms. It is the challenge of physicians, therapists and coaches involved in the care of athletes to know the symptoms of concussion, recognise them when they occur and apply basic neuropsychological testing to help detect this injury. It is, therefore, recommended to be familiar with one grading system and use it consistently, even though it may not be scientifically validated. Then good clinical judgement and the ability to recognise post concussion signs and symptoms will assure that an athlete never returns to play while symptomatic. PMID- 11475326 TI - Characterization of four extensin genes in Arabidopsis thaliana by differential gene expression under stress and non-stress conditions. AB - From Arabidopsis thaliana we isolated four different cDNAs that encode extensins, a family of cell-wall hydroxyproline-rich glycoproteins (HRGPs). Putative proteins (AtExt2-5) contained one open reading frame and characteristic Ser (Pro)4 sequences organized in a high-order repetitive motif. AtExt2-5 genes were strongly expressed during rehydration after dehydration. They were also expressed after treatment with various amino acids. In particular, AtExt3 and five mRNAs were abundantly accumulated after treatment with L-Ser, Hyp, and L-Pro, which are major components of extensin proteins. The AtExt transcripts were strongly expressed in root tissues of both unbolted and bolted plants. The transcripts of AtExt2, 3, and 5 were also detected in the lower stem and flower buds, and that of AtExt4 was detected in bolted flowers. Therefore, we suggest that these four AtExt genes are novel extensin genes in A. thaliana, because the expression of atExt1, which has already been isolated from A. thaliana, was different from these. PMID- 11475327 TI - A novel bacterial gene-finding system with improved accuracy in locating start codons. AB - Although a number of bacterial gene-finding programs have been developed, there is still room for improvement especially in the area of correctly detecting translation start sites. We developed a novel bacterial gene-finding program named GeneHacker Plus. Like many others, it is based on a hidden Markov model (HMM) with duration. However, it is a 'local' model in the sense that the model starts from the translation control region and ends at the stop codon of a coding region. Multiple coding regions are identified as partial paths, like local alignments in the Smith-Waterman algorithm, regardless of how they overlap. Moreover, our semiautomatic procedure for constructing the model of the translation control region allows the inclusion of an additional conserved element as well as the ribosome-binding site. We confirmed that GeneHacker Plus is one of the most accurate programs in terms of both finding potential coding regions and precisely locating translation start sites. GeneHacker Plus is also equipped with an option where the results from database homology searches are directly embedded in the HMM. Although this option does not raise the overall predictability, labeled similarity information can be of practical use. GeneHacker Plus can be accessed freely at http://elmo.ims.u-tokyo.ac.jp/GH/. PMID- 11475328 TI - Control and prevention of rubella: evaluation and management of suspected outbreaks, rubella in pregnant women, and surveillance for congenital rubella syndrome. AB - Outbreaks of rubella continue to occur in the United States despite widespread use of the measles-mumps-rubella (MMR) vaccine. Throughout the mid- to late 1990s, rubella outbreaks were characterized by increased numbers of cases among adults born in countries that do not have or have only recently instituted a national rubella vaccination program. To address this change in disease epidemiology, CDC's National Immunization Program (NIP) developed the following recommendations in conjunction with public health officials in the field. Public health officials should implement appropriate responses to reports of suspected rubella to determine if an outbreak exists, evaluate its scope, and implement appropriate control measures. Health-care providers should be aware of the need for rubella prevention and control among women of childbearing age and of the appropriate follow-up for pregnant women exposed to rubella. Comprehensive surveillance for congenital rubella syndrome should begin during a rubella outbreak. PMID- 11475329 TI - Increased expression of metallothionein in inflammatory bowel disease. AB - OBJECTIVE: Metallothioneins (MT) are cytoprotective against the damaging effects of oxygen-derived free radicals. Therefore MT may be involved in defence mechanisms to counter Crohn's disease (CD) and ulcerative colitis (UC). MATERIALS: 107 routinely processed tissue samples from 22 patients with CD and 48 patients with UC were tested with the monoclonal anti-MT antibody E9. METHODS: Immunohistochemistry was used to assess MT staining in a semiquantitative manner. Chi-square test was used for statistical analysis. RESULTS: MT overexpression was found in the fibroblasts of all ulcerative and/or fissural lesions in UC and CD. MT overexpression in intestinal epithelial cells of 40% of UC and CD lesions correlated significantly with the grade of inflammation. CONCLUSIONS: MT immunoreactivity in fibroblasts supports a protective role for MT in inflammatory bowel disease. It remains unclear whether MT overexpression in epithelial cells is also important in this protection. PMID- 11475330 TI - Hyaluronan molecular weight and polydispersity in some commercial intra-articular injectable preparations and in synovial fluid. AB - OBJECTIVE AND DESIGN: Hyaluronan is the major non-proteinaceous component of joint synovial fluid and is responsible for the unique rheological and biological properties of this medium. In joint arthropathies the molecular weight and concentration of hyaluronan may change, thereby influencing joint physiology and function. Intra-articular administrated hyaluronan derived from a number of sources, has been used for the treatment of osteoarthritis, however, there is limited information on the molecular weight and polydispersity of these various commercial preparations. The objective of this study was to develop an accurate, convenient method by which the molecular weight and polydispersity of hyaluronan may be determined and then applied to characterise the hyaluronan in synovial fluid. MATERIALS AND METHODS: Characterisation of the molecular parameters of hyaluronan of different origins and in ovine synovial fluid was accomplished using a multi-angle laser-light scattering (MALLS) detector coupled to a gel permeation chromatography (GPC) system, fitted with an automatic sample injector. CONCLUSION: Seven commercially available hyaluronan preparations of reported molecular weight were analysed. The weight average molecular weight (Mw) and number average molecular weight (Mn) values obtained for 6 of the 7 preparations using the MALLS-GPC system were in good agreement with the reported values. The abnormally low values for the exception suggested that degradation of hyaluronan had occurred. The MALLS-GPC technique was then used to determine the molecular characteristics of the endogenous hyaluronan in normal ovine synovial fluids. While the Mws ranged from less than 1 x 10(6) Da to 7 x 10(6) Da the majority were between 1-3 x 10(6) Da. [mean Mw = 2.42 x 10(6), mean Mn = 2.21 x 10(6) Da]. The effects of freezing and thawing synovial fluid upon molecular weight of hyaluronan were also investigated and were found to diminish both Mz and Mw values. PMID- 11475331 TI - Mouse histamine N-methyltransferase: cDNA cloning, expression, gene cloning and chromosomal localization. AB - OBJECTIVE: Histamine N-methyltransferase (HNMT) catalyzes the Ntau-methylation of histamine. We set out to clone a mouse liver HNMT cDNA and the mouse HNMT gene as steps toward characterizing molecular genetic mechanisms involved in the regulation of this important histamine-metabolizing enzyme. DESIGN: A PCR-based strategy was used to clone both the mouse HNMT cDNA and the gene encoding that cDNA, Hnmt. The cDNA was used both to express recombinant mouse HNMT and to determine the chromosomal localization of Hnmt. RESULTS: The mouse liver HNMT cDNA was 1657 bp in length with an 888 bp open reading frame (ORF) that encoded a 296 amino acid protein with a predicted Mr value of approximately 32.5 kDa. The amino acid sequence of the encoded protein was 84% identical to that of human kidney HNMT. Mouse HNMT was expressed in COS-1 cells, and its apparent Km values for histamine and S-adenosyl-L-methionine (Ado-Met), the two cosubstrates for the reaction, were 5.3 and 5.8 microM, respectively. The mouse HNMT gene, Hnmt, spanned approximately 25 kb and had 7 exons. Its structure differed from that of the human gene primarily by the presence of an additional exon at the 5' terminus. Hnmt mapped to mouse chromosome 2 in an area of conserved synteny to human chromosome 2q, the location of the human gene (2q22) on the basis of fluorescence in situ hybridization. CONCLUSIONS: Cloning and functional characterization of the mouse HNMT cDNA and gene will now make it possible to study in the mouse molecular genetic mechanisms involved the regulation of this important histamine-metabolizing enzyme. PMID- 11475332 TI - Mechanisms of allergen- and LPS-induced bone marrow eosinophil mobilization and eosinophil accumulation into the pleural cavity: a role for CD11b/CD18 complex. AB - OBJECTIVE: The mechanisms involved in bone marrow eosinophil emigration and recruitment to inflammatory sites are not fully understood. The involvement of CD11b/CD18 in marrow eosinophil release induced by lipopolysaccharide (LPS) or allergen was investigated in mice. METHODS: Eosinophil and neutrophil counts in the pleural cavity, blood and bone marrow were performed at different time intervals after the intrathoracic injection of LPS (250 ng/cavity) or ovalbumin (OVA, 12 microg/cavity; into actively sensitized mice) and compared to anti CD11b/CD 18 (5C6, 1 mg/mouse) or anti-IL-5 (TRFK-5, 500 microg/kg) treated mice. RESULTS: LPS induced local eosinophil influx, that peaked within 24 h and that was preceded by a decrease in marrow eosinophils at 4 h. Antigenic challenge induced a decrease in marrow eosinophils within 4 h, followed by a long lasting pleural eosinophil accumulation and a persistent increase in marrow eosinophil numbers. Pretreatment with anti-CD11b/CD18 abolished LPS-induced neutrophil and eosinophil accumulation in the pleural cavity at 4 and 24 h, respectively. This pretreatment failed to modify neutrophil emigration from bone marrow, but significantly inhibited marrow eosinophil release at 4 h post-LPS or OVA challenge. Anti-IL-5 pretreatment failed to inhibit LPS-induced pleural eosinophil accumulation and mobilization from bone marrow, but it abolished allergen-induced effects, indicating a role for IL-5 in marrow eosinophil mobilization induced by antigen, but not by LPS challenge. CONCLUSIONS: Our results suggest that eosinophil migration induced by antigen or LPS into the pleural cavity is preceded by bone marrow eosinophil release through a mechanism that depends on CD11b/CD18. PMID- 11475333 TI - Effect of imidazo[1,2-a]pyrimidine derivatives on leukocyte function. AB - OBJECTIVE AND DESIGN: A series of six imidazo[1,2-a]pyrimidine (IP) derivatives were evaluated for their effects on leukocyte functions in vitro as well as on the inflammatory response induced by zymosan in the mouse air pouch. MATERIALS AND SUBJECTS: Human neutrophils and murine peritoneal macrophages were used for in vitro assays. Mouse air pouch was performed in Swiss mice. TREATMENT: Test compounds were incubated with either human neutrophils or mouse peritoneal macrophages at concentrations not showing cytotoxic effects. For in vivo experiments, IPs were injected into the air pouch. METHODS: Elastase and myeloperoxidase release, superoxide generation and LTB4 production were assayed in human neutrophils treated with different stimuli. Mouse peritoneal macrophages were stimulated with lipopolysaccharide (LPS), followed by determination of nitrite and PGE2 levels in supernatants. Zymosan was injected into six days-old mouse air pouches. Dunnett's t-test was employed for statistical analysis. RESULTS: All IPs inhibited human neutrophil degranulation with IC50 values in the microM range. IP-1, IP-2 and IP-5 also decreased superoxide generation. In LPS stimulated macrophages, IP-4 and IP-6 inhibited nitrite production with a moderate reduction in PGE2 generation. In addition, these two compounds at 100 nmol/pouch inhibited leukocyte migration and LTB4 levels in the exudate of the mouse air pouch. CONCLUSIONS: Imidazo[1,2-a]pyrimidines are a class of compounds with antiinflammatory potential. PMID- 11475334 TI - Regulation of 15-lipoxygenase isozymes and mucin secretion by cytokines in cultured normal human bronchial epithelial cells. AB - OBJECTIVE AND DESIGN: To study the effects of IL-4, IL-13, IL-1beta, IL-8 and TNFalpha on 15-lipoxygenase(15-LO) isozyme expression and mucin secretion in normal human bronchial epithelial cells. MATERIAL AND METHODS: The effects of IL 4, IL-13, IL-1beta, IL-8 and TNFalpha on 15-LO isoenzyme mRNA and protein expression, total 15-LO enzyme activity and mucin secretion were examined in cultures of normal human bronchial epithelial cells. In addition, in order to determine whether the observed effects on mucin secretion were due to lipoxygenase (LO) products, the effect of the non-selective LO inhibitor nordihydroguaiaretic acid was examined. RESULTS: IL-4 and IL- 13 selectively enhanced 15-LOa mRNA and protein levels, and total 15-LO enzyme activity. In contrast, no changes were observed in 15-LOb mRNA or protein levels. IL-4 and IL 13 both reduced mucin secretion in this cell type, however the non-selective LO inhibitor nordihydroguaiaretic acid had no effect on this action of IL-4. CONCLUSIONS: These data demonstrate that IL-4 and IL-13 selectively regulate the expression of the 15-LOa isozyme. However, 15LOa products do not mediate the IL-4 induced reduction in mucin secretion observed in this cell type. PMID- 11475335 TI - The analysis of nonsteroidal antiinflammatory drug selectivity in prostaglandin G/H synthase (PGHS)-null cells. AB - OBJECTIVES: Nonsteroidal antiinflammatory drugs (NSAIDs) that are more selective for PGHS-2 maintain their antiinflammatory properties but exhibit fewer unfavorable gastrointestinal side effects. To evaluate the usefulness of the murine PGHS-null cell system in analyzing PGHS-2 selective inhibitors, we tested PGHS-2 non-selective NSAIDs and two PGHS-2 specific compounds using either endogenous or exogenous sources of substrate, arachidonic acid. MATERIALS AND METHODS: A whole-cell assay system for testing the efficacy of PGHS isozyme specific inhibitors using murine PGHS-1 or PGHS-2-null fibroblast cell lines derived from lung tissues of PGHS-2(-/-) and PGHS-(-/-) mice, respectively, was employed. PGHS-1 and PGHS-2 null cell lines were exposed to three widely used NSAIDs, ibuprofen, indomethacin and aspirin, and two PGHS-2 specific inhibitors, MK-966 (rofecoxib) and NS-398. Excess arachidonic acid (AA) was provided externally and internally via calcium ionophore A23187. PGHS-1 and PGHS-2 activity were determined by measuring the prostaglandin E2 production by radioimmunoassay. IC50 and IC80 values of each compound were determined from the reduction of PGE2 levels as a measure of inhibition of existing PGHS isozyme in the PGHS-null cells. RESULTS: In our murine PGHS-null cell systems, both PGHS-1 and PGHS-2 null cells can use both externally provided AA and endogenous, A23187 derived AA. Both NS-398 and MK-966 were potent inhibitors and demonstrated strong selectivity for PGHS-2. Among the non-selective NSAIDs, based on the PGHS-2 IC50/PGHS-1 IC50 ratio ranking, ibuprofen is more selective for PGHS-2 than indomethacin while aspirin is the least selective inhibitor regardless of the arachidonic acid source. Indomethacin and MK-966 IC50 values for PGHS-2 were in the range of 10(-9)-10(-8) M while the IC50 values for aspirin were in the range of 10(-5) M. There were differences in the ranking of indomethacin and ibuprofen when the IC80 ratios were used. CONCLUSION: Here, we report on a whole cell assay system for testing the efficacy of PGHS isozyme-specific inhibitors using murine PGHS-1 or PGHS-2-null cell lines. This system, using cells that express either PGHS-1 or PGHS-2, offers a convenient and reliable method to determine IC50 and IC80 values of the two PGHS isoforms, entirely independent of each other, in the same cell type. The results of our evaluation using a panel of NSAIDs, both PGHS 2 selective and non-selective inhibitors, correlate well with previously published clinical and laboratory data, demonstrating the usefulness of the whole cell assay system described here. PMID- 11475336 TI - Increased expression of G12 and G13 proteins in bronchial smooth muscle of airway hyperresponsive rats. AB - OBJECTIVE: To obtain information on the activation pathway of the monomeric G protein, RhoA, in bronchial smooth muscle, the expression of G alpha12 and G alpha13 in bronchial smooth muscle of the rat was determined. The levels of these G proteins were also compared between antigen-induced airway hyperresponsive and normal control groups. METHODS: Actively sensitized rats were repeatedly challenged by antigen inhalation. Twenty-four hours after the final antigen challenge, membrane preparations of bronchial smooth muscles were prepared. Immunoblottings were performed, and the density ratios of G alpha12/beta-actin and G alpha13/beta-actin were calculated to quantify the levels of these G protein alpha subunits. RESULTS: Both G alpha12 and G alpha13 proteins were expressed in rat bronchial smooth muscle. The levels of bronchial G alpha12 and G alpha13 proteins in the repeatedly antigen challenged rats were significantly increased as compared with those in control animals; the magnitude of upregulation in the airway-hyperresponsive group was 89% and 68% in the control group, respectively. CONCLUSION: G alpha12 and G alpha13 proteins were expressed in rat bronchial smooth muscle. Considering the probable involvement of G12 and G13 proteins in Ca2+ sensitization through Rho protein, the augmented expression of such G proteins after repeated antigen challenge may be responsible for the hyperresponsiveness of bronchial smooth muscle contraction in rats. PMID- 11475337 TI - Vancomycin for the initial therapy of peritonitis: don't throw out the baby with the bathwater. PMID- 11475338 TI - Guidelines by an Ad Hoc European Committee for Elective Chronic Peritoneal Dialysis in Pediatric Patients. PMID- 11475339 TI - Structural and functional alterations of the peritoneum after prolonged exposure to dialysis solutions: role of aminoguanidine. AB - OBJECTIVE: The effect of long-term use of high glucose dialysate on peritoneal structure and function, and its relation with accumulation of advanced glycosylation end-product (AGE) in the peritoneum was investigated in this study. METHODS: Dialysates with 4.25% glucose were injected into the peritoneal cavity of normal rats for 12 weeks without (PD, n = 7) and with (1 g/L, PD+AG, n = 7) aminoguanidine in their drinking water. Rats not having intraperitoneal (IP) injection served as control (n = 9). After 12 weeks of IP injection, a 2-hour peritoneal equilibration test (PET) was performed using 30 mL 4.25% glucose dialysate. Intraperitoneal volume (IPV), dialysate-to-plasma urea ratio at 2 hours (D/P2), the ratio of dialysate glucose at 2 hours to initial dialysate glucose (D2/D0), and the peritoneal fluid absorption rate (Qa) were evaluated. After the PET, samples of the parietal peritoneum were taken for hematoxylin and eosin (H&E) staining and immunohistochemical staining for AGE. RESULTS: The IPV and D2/D0 glucose were significantly lower and Qa and D2/P2 urea significantly higher in the PD group than in the control group. Aminoguanidine reversed in part the changes in IPV and D2/P2 urea in the PD group; it had no effect on Qa and D2/D0 glucose. The H&E staining showed a linear mesothelial lining with negligible cells and capillaries in the narrow submesothelial space in the control group. Mesothelial denudation and submesothelial infiltration of monocytes and capillary formation were observed in the PD group. Mesothelial denudation was relatively intact in the PD+AG group compared with the PD group. Submesothelial monocyte infiltration and capillary formation in the PD+AG group were not as prominent as in the PD group. Positive AGE staining was found in the submesothelial space, vascular walls, and endomysium in the PD group, while it was markedly attenuated in PD+AG group and negligible in the control group. CONCLUSION: Long-term use of high glucose solutions induced peritoneal AGE accumulation and mesothelial denudation, and increased peritoneal permeability and peritoneal fluid absorption rate. Inhibition of peritoneal AGE accumulation prevented those functional and structural damages to the peritoneum. PMID- 11475340 TI - A new titanium fiber mesh-cuffed peritoneal dialysis catheter: evaluation and comparison with a Dacron-cuffed tenckhoff catheter in goats. AB - OBJECTIVE: We report the development of a new peritoneal dialysis catheter. The purpose of the current study was to evaluate tissue response to this catheter. DESIGN: In our catheter design, the catheter's cuffs are made from sintered titanium fiber mesh instead of the common Dacron material. The catheters were implanted in goats. During the study period, the catheter exit sites were observed once every 2 weeks. After 14 weeks, the catheters and surrounding tissues were processed for histology and analyzed. RESULTS: We observed that catheters with titanium cuffs performed significantly better, compared with a commercial catheter design. Histology showed that titanium fibers evoked a lesser inflammatory response compared with Dacron fibers. In addition, the capsule surrounding the titanium cuff was significantly thinner. CONCLUSIONS: Supported by the obtained results, we conclude that the use of Dacron velour for anchorage of percutaneous catheters has to be reconsidered. Titanium fiber mesh has a great potential for application in percutaneous catheters. PMID- 11475341 TI - Peritoneal dialysis clearance can replace residual renal function. AB - OBJECTIVE: There is controversy whether increasing peritoneal clearance effectively substitutes for declining residual renal function. We studied the impact of renal and peritoneal clearances on outcome, controlling for comorbidity. DESIGN: Registry database. SETTINGS: Four dialysis centers. PATIENTS: Incident peritoneal dialysis patients. METHODS: Data were collected prospectively on 90 incident patients between 1991 and 1999. At the end of their first year on peritoneal dialysis, patients were divided into groups based on the first year's clearance results: group 1 (n = 62) had weekly Kt/W greater than or equal to 2.0 and creatinine clearance (CCr/1.73 m2) greater than or equal to 60 L throughout the first year; group 2 (n = 28) fell below these targets due to loss of residual renal function and then reached targets due to prescription change. MAIN OUTCOME MEASURES: Patient and technique survival. RESULTS: Both groups were similar in baseline characteristics except age (57 years vs 49 years, p = 0.02) and initial albumin (34.4 g/L vs 37.5 g/L, p = 0.001). One-year patient survival after grouping was similar in both groups (86.3% vs 80.9%, p = 0.72). Cox proportional hazard model, controlling for comorbidity, did not show "group" to be a significant predictor of outcome (p = 0.96). One-year technique survival after grouping was similar in both groups (77.3% vs 83.2%, log rank p = 0.89). For technique failure, Cox proportional hazard model showed peritonitis (p = 0.004) to be the only significant predictor of worse outcome. CONCLUSIONS: Peritoneal dialysis patients with improved clearances due to prescription changes had survival comparable to patients who never fell below target. This suggests that loss of residual renal function may be replaced by increasing peritoneal dialysis clearance. A large multicenter trial to study this important question further is needed. PMID- 11475342 TI - Icodextrin degradation products in spent dialysate of CAPD patients and the rat, and its relation with dialysate osmolality. AB - OBJECTIVE: Peritoneal dialysis (PD) with a 7.5% icodextrin-containing dialysis solution provides prolonged ultrafiltration compared with glucose-based dialysis solutions. Colloid osmosis is the most likely mechanism, but studies in rats suggest it is caused by an increase in osmolality due to degradation of icodextrin. Therefore, human spent dialysate was analyzed with high-performance liquid chromatography (HPLC) using gel permeation size-exclusion chromatography. An increasing peak (with a low molecular weight, < 1000 Da) was observed during the dwell. The aim of this study was to quantitate breakdown products of icodextrin (which could explain this peak) and investigate whether there was a relationship with dialysate amylase concentration and dialysate osmolality. DESIGN: Long-dwell effluents (dwell time 9.15- 14.30 hours) obtained from 12 PD patients using a 7.5% icodextrin solution during the night were analyzed. The following icodextrin breakdown products were measured: maltotetraose (G4), maltotriose (G3), maltose (G2), and glucose (G1). In 6 of these patients, the sugars maltoheptaose (G7), maltohexaose (G6), and maltopentaose (G5) were also determined in both effluent and plasma. In addition, G4, G3, G2, and G1 were measured in four Wistar rats during a 6-hour dwell study. RESULTS: In the human studies, the median distribution of the sugars in the effluent was G4,6.7%; G3,16.5%; G2, 23.1%; and G1, 53.5%. The osmolality in spent dialysate ranged between 288 and 326 mOsm/kg H2O. The median contribution of the sugars G2 - G4 was 5.4 mOsm/kg H2O. No correlation was present between dialysate osmolality and duration of the dwell (r= -0.04, p= 0.91); nor was there a relation between the concentration of G2 and duration of the dwell (r = 0.50, p = 0.10). No relationship was found between the amount of amylase and the concentration of G2 in the effluent (r = 0.49, p = 0.10), nor between the total concentration of the sugars G2 - G4 in the spent dialysate and dialysate osmolality (r = -0.31, p = 0.33). However, a strong correlation was seen between urea concentration and osmolality (r= 0.85, p < 0.001), and also between sodium concentration and dialysate osmolality in the spent dialysate (r = 0.92, p < 0.0001). The levels of the sugars G2, G3, and G4 in effluent were higher than in unused dialysate, but lower than or similar to plasma levels. Concentrations of the sugars G5, G6, and G7 were lower in spent dialysate than in unused dialysate, and higher than in plasma. In the rat study, dialysate osmolality increased with the duration of the dwell. A clear relationship was present between osmolality and concentration of the sugars G2 - G4 in the effluent. The median amount of amylase in the effluent was 1252 U/L. CONCLUSION: A 7.5% icodextrin-based dialysis solution used during the long exchange caused only a slight increase in dialysate osmolality in humans. The osmolality at the end of the dwell in the human situation was dependent mainly on concentrations of the small solutes urea and sodium in the effluent. The contribution of icodextrin degradation products was marginal. In the rat, however, a clear relationship was present between osmolality and icodextrin degradation products in spent dialysate, explaining the increased dialysate osmolality at the end of the dwell. The difference between the two species can be explained by the very high amylase concentrations in the rat, leading to a rapid degradation of icodextrin. The rat is therefore not suitable to study peritoneal fluid kinetics using icodextrin as an osmotic agent. PMID- 11475343 TI - Effects of peritoneal dialysis with an overnight icodextrin dwell on parameters of glucose and lipid metabolism. AB - OBJECTIVE: To examine whether a reduced daily glucose load by overnight application of the less-absorbed glucose polymer icodextrin would have favorable effects on lipid profiles of continuous ambulatory peritoneal dialysis (CAPD) patients. STUDY DESIGN: Randomized crossover study with two subsequent periods of 6 weeks. SETTING: Home PD unit of a secondary-care hospital. PATIENTS: Twenty-one nondiabetic CAPD patients (15 male, 6 female; mean age 50.3+/-11.8 years). INTERVENTION: Participants were randomly assigned to receive an overnight dwell with either standard glucose solution or with a 7.5% icodextrin-containing solution. MAIN OUTCOME MEASURES: Relation between reduction in the total amount of intraperitoneal infused glucose and parameters of glucose (plasma glucose, insulin, and HbA1C) and lipid metabolism [free fatty acids, plasma lipids, lipoproteins, and low density lipoprotein (LDL) subfraction profile]. RESULTS: After the icodextrin dwells, a reduction of plasma total cholesterol (from 5.43+/ 0.85 to 4.86+/-0.70 mmol/L, p < 0.001) and LDL cholesterol (from 3.38+/-0.87 to 2.93+/-0.73 mmol/L, p = 0.001) was observed. Also, high density lipoprotein (HDL) cholesterol (from 0.95+/-0.27 to 0.90+/-0.24 mmol/L, p = 0.029) was reduced, but the plasma total cholesterol-to-HDL ratio remained similar. Plasma free fatty acids and triglyceride levels tended to decrease (from 0.16+/-0.10 to 0.13+/-0.08 mmol/L, p= 0.06, and from 2.14+/-1.96 to 1.92+/-1.03 mmol/L, respectively). Evaluation of LDL subfraction profiles after ultracentrifugation showed a more buoyant LDL subfraction profile with fewer dense LDL particles in 6 patients and no changes in 14 patients after icodextrin. The effects on lipids were not accompanied by a decrease in fasting plasma glucose (from 5.76+/-1.29 to 5.86+/ 0.80 mmol/L) or insulin levels (from 19.5+/-14.4 to 20.3+/-13.0 mU/L). CONCLUSION: These results suggest a beneficial effect on lipid profiles of CAPD patients with the use of an overnight dwell with icodextrin. PMID- 11475344 TI - Association of hypertriglyceridemia and insulin resistance in uremic patients undergoing CAPD. AB - OBJECTIVE: Hyperlipidemia is frequently encountered in uremic patients and may be worsened by continuous ambulatory peritoneal dialysis (CAPD) treatment. The lipid abnormalities in these patients may be multifactorial. Insulin resistance (or its compensatory hyperinsulinemia) is commonly observed in uremic patients, but its association with hyperlipidemia in these patients has not been studied. PATIENTS AND METHODS: Lipid profiles of 35 nondiabetic nonobese patients undergoing CAPD for more than 1 year (mean 52.3 months) were studied. Current laboratory data and parameters related to peritoneal dialysis (PD) within the previous 3 months were recorded. After overnight fasting and interruption of PD, an oral 75-g glucose tolerance test (OGTT) was examined. RESULTS: After CAPD treatment for more than 12 months, these patients had higher serum triglyceride (TG) (p = 0.001) and total cholesterol (p = 0.0058) levels than their values before commencing CAPD. Twelve of 14 patients with serum TG higher than 200 mg/dL (high-TG) were diagnosed de novo, in contrast with only 1 patient diagnosed of de novo hypercholesterolemia (total cholesterol > 240 mg/dL). There was no difference in age, gender, body mass index (BMI), duration of PD treatment, serum albumin, hematocrit, intact serum parathyroid hormone (iPTH), peritoneal glucose load, solute transport, or weekly Kt/V urea between normal-TG and high-TG patients. After adjusting for age, gender, BMI, weekly Kt/V urea, and iPTH, the high-TG patients had higher levels of area under the curve for glucose (AUC(Glu)), area under the curve for insulin (AUC(Ins)), and AUC(Ins)/AUC(Glu) ratios (F = 10.63, 10.14, and 8.65; p = 0.0029, 0.0035, and 0.0065, respectively), indicating that the high-TG patients were more insulin resistant. There were 24 patients with normal glucose tolerance (NGT), and 11 patients with impaired glucose tolerance (IGT). The IGT group had higher serum TG (F = 10.43, p = 0.003) and total cholesterol (F = 8.05, p = 0.009) than the NGT group, after adjusting for BMI, duration of CAPD treatment, peritoneal glucose load, solute transport, serum albumin, and lipid levels before PD treatment. TheTG levels after CAPD treatment were positively correlated with AUC(Glu), AUC(Ins), and AUC(Ins)/AUC(Glu) ratio (r = 0.48, 0.53, and 0.49; p = 0.0037, 0.001, and 0.0028, respectively). CONCLUSIONS: These results indicate that insulin resistance is an important factor in the development of hypertriglyceridemia in CAPD patients. PMID- 11475345 TI - Experience with a large dose (500 mg) of intravenous iron dextran and iron saccharate in peritoneal dialysis patients. AB - OBJECTIVE: To compare efficacy in anemia correction and side effects of large doses of intravenous (IV) iron dextran and iron saccharate preparations in peritoneal dialysis (PD) patients. SETTING: Tertiary-care teaching hospital of University of Toronto. DESIGN: Retrospective analysis of 379 PD patients who attended PD clinics in past 5 years. Of these 379 patients, 62 were selected to receive IV iron based on ferrokinetic markers of iron deficiency, noncompliance to or ineffectiveness of oral iron, or increased erythropoietin (EPO) requirement. INTERVENTION: Sixty-one patients received two IV iron injections of 500 mg each, 1 week apart, 33 patients received iron dextran, 23 received iron saccharate, and 5 received both iron dextran and iron saccharate. One patient developed anaphylaxis to a test dose of iron dextran and was excluded from further therapy. Blood samples were collected before and 3 and 6 months after iron infusions. RESULTS: At 3 months, the group's average hemoglobin rose from 98.3+/-18.3 g/L to 110.6+/-16.4 g/L (p < 0.0001). Ferritin rose from 104.9+/ 115.4 microg/L to 391.5+/-294.1 microg/L (p < 0.0001), and transferrin saturation from 0.17+/-0.07 to 0.26+/-0.19 (p < 0.0001). Erythropoietin requirements fell from 7278.7 IU/week to 5900 IU/week (p < 0.01). Five of the 34 patients who received iron dextran developed minor side effects and 1 patient had anaphylaxis to the test dose. Of the 23 patients who received iron saccharate, 1 had an anaphylactic reaction and 2 had transient chest pain, which subsided without therapy. Overall, there were more side effects with iron dextran (7.4% of injections) compared to the iron saccharate group (4.3% of injections), but this difference was statistically insignificant. Although statistically insignificant, there was an increase in the number of peritonitis episodes during the 6 months after IV iron infusion, especially with iron dextran, compared to the peritonitis episodes during the 6 months before iron infusions. CONCLUSION: Our study indicates that IV iron in PD patients is effective in restoring iron stores and in decreasing EPO requirements. One anaphylactic reaction occurred in each group. Our data suggest that as much caution be exercised with iron saccharate as with iron dextran. The slight trend toward increased peritonitis rates after iron infusions needs to be investigated in a larger group of patients. PMID- 11475346 TI - Polymicrobial outbreak of intermittent peritoneal dialysis peritonitis during external wall renovation at a dialysis center. AB - OBJECTIVE: To investigate an outbreak of peritonitis in intermittent peritoneal dialysis (IPD) patients. DESIGN: An outbreak investigation was performed to identify the etiology of the polymicrobial outbreak, and a retrospective case control study was conducted to assess the risk factors for development of peritonitis. SETTING: Renal dialysis center. PATIENTS: Ten episodes of peritonitis occurred in 8 of 61 patients over a 6-month period in which 669 IPD procedures were analyzed. INTERVENTIONS: Field visit to renal dialysis center to examine the entire IPD procedure, inspect the hospital environment, and perform air bacterial count. MAIN OUTCOME MEASURES: The environmental factors and risk factors contributing to the polymicrobial peritonitis outbreak in IPD patients. The incidence of IPD peritonitis was determined before and after interventions. RESULTS: The causative organisms included Acinetobacter baumanii (6), Stenotrophomonas maltophilia (2), Pseudomonas aeruginosa (1), Candida albicans (1), C. tropicalis (1), Enterococcus (3), and Enterobacteriaceae (2). Four episodes of peritonitis involved infection by more than one organism. Air sampling of the environment detected a median of 110 colony forming units of bacteria per cubic meter of air, 10% of which were found to be Acinetobacter baumanii. The source of this polymicrobial outbreak was attributed to the bamboo scaffolding structure covering the external wall of the hospital during renovation. A retrospective case-control study indicated that the absence of the flush-before-fill step was a risk factor for development of peritonitis. CONCLUSION: In addition to invasive aspergillosis in transplant or oncology patients, Acinetobacter peritonitis in dialysis patients should be considered another microbial cause of outbreak associated with hospital renovation. PMID- 11475347 TI - The influence of demographic factors and modality on loss of residual renal function in incident peritoneal dialysis patients. AB - OBJECTIVE: To determine whether gender, race, diabetes, peritoneal dialysis (PD) modality, and comorbid conditions influence loss of residual renal function (RRF). DESIGN: Retrospective study of incident PD patients, using database of prospectively collected demographic, laboratory, and clearance data. SETTING: Peritoneal Dialysis Registry of the University of Pittsburgh Medical Center. PATIENTS: The study included 184 continuous ambulatory PD and automated PD patients who had at least two 24-hour urine collections for glomerular filtration rate (GRF) between April 1991 and March 2000. 836 urine collections were analyzed. OUTCOME MEASURES: Loss of RRF was defined as the slope of the decline in GFR as measured by the average of creatinine and urea clearances in 24-hour urine collections. Stepwise forward regression was used to identify demographic and laboratory factors associated with loss of GFR. Spearman correlations were used to assess the significance of associations. RESULTS: The median rate of decline of renal function was -0.17 mL/minute/month. Gender, race, diabetes, automated PD, peritoneal equilibration test, protein equivalent of nonprotein nitrogen appearance normalized to body surface area, and serum albumin did not predict loss of RRF. Cardiac disease was the only variable affecting decline of RRF (p = 0.045). CONCLUSION: Modality of PD and patient demographic factors do not contribute to the rate at which RRF is lost in incident PD patients. Additional study of the factors contributing to the decline and maintenance of RRF is needed. PMID- 11475348 TI - A comparison of quality of life of patients on automated and continuous ambulatory peritoneal dialysis. AB - OBJECTIVE: Data on health-related quality of life (HRQOL) of automated peritoneal dialysis (APD) patients are scarce. The objectives of this study were (1) to explore HRQOL of APD patients and compare it with HRQOL of continuous ambulatory peritoneal dialysis (CAPD) patients and a general population sample, and (2) to study the relationship between HROOL assessment outcomes and background variables. DESIGN: Home interviews of APD and CAPD patients. HRQOL, social demographic, clinical, and treatment-related background data were collected at the interview and from patient charts. Multiple regression analysis and logistic regression analysis were used to study the relationship of HRQOL assessment outcomes with background variables. SETTING: Sixteen Dutch dialysis centers. PATIENTS: Convenience sample of 37 APD patients and 59 CAPD patients matched for total time on dialysis. MAIN OUTCOME MEASURES: Four HRQOL instruments: Short-Form 36, EuroQol EQ-5D, Standard Gamble, and Time Trade Off. RESULTS: Physical functioning of both APD and CAPD patients was impaired compared with the general population; mental functioning was not different. In multivariate analyses, the mental health of APD patients was found to be better than that of CAPD patients. In addition, APD patients were less anxious and depressed than CAPD patients. With respect to physical aspects of HRQOL and role-functioning, no differences were observed between APD and CAPD patients. Other variables to explain HRQOL assessment outcomes were age, the number of comorbid diseases, and primary kidney disease. CONCLUSIONS: HRQOL of APD patients is at least equal to HRQOL of CAPD patients. PMID- 11475349 TI - Injury to the inferior epigastric artery complicating percutaneous peritoneal dialysis catheter insertion. PMID- 11475350 TI - A case of candida peritonitis followed by mediastinitis after esophageal perforation in a peritoneal dialysis patient. PMID- 11475351 TI - Intravenously administered erythropoietin may cause a direct increase in blood pressure in CAPD patients. PMID- 11475352 TI - Concerns regarding recommendations for the treatment of CAPD peritonitis. PMID- 11475353 TI - On the recent recommendations of the Ad Hoc Advisory Committee on Peritonitis Management--or should ceftazidime be used as initial empiric therapy? PMID- 11475354 TI - Peritoneal reaction to icodextrin in a female patient on CAPD. PMID- 11475355 TI - Literature: May-June 2001. PMID- 11475356 TI - Arterial smooth muscle cell proliferation on a novel biomimicking, biodegradable vascular graft scaffold. PMID- 11475357 TI - Liposomes: applications in medicine. AB - Liposomes are spherical lipid bilayers from 50 nm to 1000 nm in diameter that serve as convenient delivery vehicles for biologically active compounds. The field of liposome research has expanded considerably over the last 30 years. It is now possible to engineer a wide range of liposomes varying in size, phospholipid composition and surface characteristics to suit the specific application for which they are intended. This paper gives an overview of the main advances in liposome research from a point of view of their applications in medicine. Aqueous contrast enhancing agents entrapped in liposomal carriers can be targeted to the liver and spleen and distinctions can be made between normal and tumorous tissue using computed tomography. Topical application of liposomes has great potential in dermatology. Liposomes have been used to deliver anticancer agents in order to reduce the toxic effects of the drugs when given alone or to increase the circulation time and effectiveness of the drugs. From the original concept of encapsulating hemoglobin in an inert shell, liposome encapsulated hemoglobin (LEH) has evolved into a fluid proven to carry oxygen, capable of surviving for reasonable periods in the circulation and amenable to large-scale production. Liposomes may be used to target specific cells by attaching amino acid fragments such as antibodies or proteins or appropriate fragments that target specific receptor sites. Liposomal DNA delivery vectors and further enhancements in the forms of LPDI and LPDII are some of the safest and potentially most versatile transfer vectors used to date. DNA vaccination and improved efficiency of gene therapy are just a few of the upcoming applications of liposomes. PMID- 11475358 TI - Xanthan gum as a carrier for controlled release of drugs. AB - Systems based on xanthan gum matrix containing 1%, 2% and 5% salicylic acid were prepared and studied as controlled release devices. Swelling of the matrix in distilled water and buffer solutions showed that the ionic strength of the liquid has a strong effect on the sorptive properties of the matrix. From the release experiments, conducted in distilled water at 37 +/- 0.5 degrees C, it was found that the drug delivery process was accomplished within the first 10 hours after immersion and salicylic acid was always released via a non-Fickian transport. The phenomenon can be described by a release exponent (n) in the area of 0.77 independently of the initial concentration of salicylic acid in the xanthan matrix. These results can be interpreted taking into consideration the dimensional and physical changes of the polymeric matrix during swelling. PMID- 11475359 TI - Influence of the selection of the suture material on the mechanical behavior of a biomaterial to be employed in the construction of implants. Part 1: Calf pericardium. AB - A hydraulic stress simulator was employed to study the mechanical behavior of the calf pericardium used in the construction of cardiac valve leaflets. One hundred eighty pairs of tissue samples were subjected to tensile testing to rupture. One of the two samples from each of 144 pairs (four series of 36 pairs each) was sutured with commercially available threads made of nylon, silk, Prolene or Gore Tex, while the other sample in each of these pairs was left unsewn. The remaining 36 pairs were employed as controls in which neither of the two samples was subjected to suturing. The sutured tissue samples showed a significant decrease in tensile strength at rupture (range: 11.81 to 26.04 MPa) when compared with unsutured samples (range: 39.38 to 87.96 MPa; p < 0.01). The application of morphological and mechanical selection criteria to maximize the homogeneity of the samples provided excellent fit with respect to the stress/strain curves. This method made it possible to carry out a predictive study of the mechanical behavior of a sutured sample, based on that observed in the corresponding unsutured fragment. The interaction of the different suture materials with the pericardial tissue was also assessed by comparing the mechanical behavior of the sutured samples with that of the control samples. At stresses of less than 0.8 MPa, samples sewn with Gore-Tex were found to show the least difference with respect to the controls, indicating that this material presented the lowest degree of interaction with the pericardium. In conclusion, the degree of the loss of resistance to tearing of the sutured samples is of no value in the selection of the optimal suture material. The selection process applied makes it possible to predict the mechanical behavior in response to suturing of a given unsewn tissue specimen by determining that of its sutured mate. The similarity between the findings in samples sewn with Gore-Tex and in the unsutured controls indicates a lesser degree of interaction between the suture material and the pericardium employed in the construction of cardiac valve leaflets. PMID- 11475360 TI - Influence of the selection of the suture material on the mechanical behavior of a biomaterial to be employed in the construction of implants. Part 2: Porcine pericardium. AB - Using a hydraulic stress simulator, the mechanical behavior of the porcine pericardium used in the construction of cardiac valve leaflets was characterized following the same procedure employed with calf pericardium in Part 1 of this study. One hundred fifty pairs of tissue samples were subjected to tensile testing to rupture. One of the two samples from each of 120 pairs (four series of 30 pairs each) was saturated with commercially available threads made of nylon, silk, Prolene or Gore-Tex, while the other sample in each of these pairs was left unsewn. The remaining 30 pairs were employed as controls in which neither of the two samples was subjected to suturing. The sutured tissue samples showed a significant decrease in tensile strength at rupture (range: 11.61 to 21.22 MPa) when compared with unsutured samples (range: 50.80 to 89.45 MPa; p < 0.01). When these results were compared with their equivalent in calf pericardium, no significant differences were observed (the mean values at rupture in calf pericardium ranged between 211.61 MPa and 26.04 MPa). Again, the application of morphological and mechanical selection criteria to ensure the homogeneity of the samples provided excellent fit with respect to the stress/strain curves. The interaction of the different suture materials with the pericardial tissue was also assessed by comparing the mechanical behavior of the sutured samples with that of the control samples. At the working stress of a cardiac valve leaflet, 0.250 MPa, samples sewn with Gore-Tex were found to show the least difference in behavior with respect to the controls, indicating that this material presented the lowest degree of interaction with the pericardium. In conclusion, the suture clearly has deleterious effects on the resistance of both calf and porcine pericardium, which showed no statistically significant differences in terms of resistance to rupture when their respective sutured or unsutured samples were compared, except in the case of porcine pericardium sewn with silk, which presented lower resistance to rupture in all the zones studied. These findings suggest that the hypothesis that porcine pericardium is less resistant is erroneous. The Gore-Tex suture also presented a lower degree of interaction with the porcine pericardium, with values similar to the working stress of a cardiac valve leaflet. This methodology and the results should be evaluated in dynamic studies, such as fatigue testing, that not only confirm the resistance of the material but establish the durability of the samples being assayed. PMID- 11475361 TI - A contact lens electrode with built-in high intensity white light-emitting diodes. A contact lens electrode with built-in white LEDs. AB - We determined the clinical usefulness of a new contact lens electrode with built in, white light-emitting diodes (LEDs) for full-field electroretinograms (ERGs). Three, high-brightness white LEDs were incorporated into a contact lens electrode and served as the source for the stimulus and the background. The stimulus intensity, stimulus duration and background illumination were regulated by a small LED control device. Maximum stimulus and background intensities were 3.9 and 3.6 log cd/m2, respectively. We successfully recorded intensity-response series for scotopic and photopic ERGs. We also recorded a duration-series for photopic ERGs, and ERGs that were comparable to the ISCEV standardized ERGs. The compactness and ease of using this system suggest that it will be clinically useful under different conditions. PMID- 11475362 TI - Eye votives from Croatian shrines. AB - This paper investigates votive gifts that people brought to sacred places in Croatia, as a gift in connection with various eye diseases. The custom of votive giving has been present in the territory of Christian Europe from ancient times until recent days. By examining the models of eyes dating from the seventeenth to the twentieth century, we came to the conclusion that they do not show any malformation, meaning that no pathological processes of eyes were shown. With their healthy form, they show the image of a healthy eye that the one who prays wishes to have. This is the opposite from the eye votives from Greek and Roman times, which were given in the ancient Esculap shrines with the desire to be healthy, because the Greek and Roman votives do show pathological processes. PMID- 11475363 TI - The incidence of negative ERG in clinical practice. AB - INTRODUCTION: A negative electroretinogram (ERG) is one in which there is a selective reduction in amplitude of the b-wave, such that it does not exceed that of the a-wave. The purpose of this study was to determine the incidence and clinical causes of negative ERGs at a tertiary referral centre. In addition, interesting and previously unreported aetiologies are described. PATIENTS AND METHODS: Retrospective review of all ERGs done at Moorfields Eye Hospital from November 1995 to December 1998 under ISCEV standard conditions. Many patients had photopic ON- and OFF-response recording in addition to conventional ISCEV Standard ganzfeld ERG. RESULTS: A total of 2,640 ERGs were performed during the study period. 128 cases (4.8%) showed a negative ERG. The causes, where a firm clinical diagnosis was possible, include X-linked juvenile retinoschisis, congenital stationary night blindness, central retinal artery occlusion, birdshot chorioretinopathy and melanoma-associated retinopathy (MAR). Unilateral negative ERG waveforms with normal fundal appearances were seen in 7 patients. Photopic ON responses could be selectively affected. CONCLUSIONS: The incidence of negative ERGs over a 34-month period presenting to a large tertiary centre was almost 5%. The presence of a negative ERG may be instrumental in demonstrating the site of visual dysfunction, with many cases showing minimal or no fundus abnormality. ON- and OFF-response recording yielded additional information regarding photopic post receptoral/phototransduction function. PMID- 11475364 TI - Multifocal ERG changes before and after macular hole surgery. AB - To evaluate the visual function of 15 eyes suffering from macular hole the multifocal ERG was used pre and postoperatively. In all the cases a successful vitrectomy has been done with macular hole closure in all the eyes. The Multifocal ERG shows an improvement of retinal response density in regions 1 and 2, even in the 2 eyes with unchanged visual acuity postoperatively. Although a relationship exist between the postoperative retinal response density and the visual acuity, the retinal response density varies between eyes with the same visual acuity. PMID- 11475365 TI - Visual evoked cortical potential can be used to differentiate between uncorrected refractive error and macular disorders. AB - The visual evoked cortical potential (VECP) is widely used to verify complaints of reduced visual performance and to identify the site of the disorder. In this study, we investigated the correlation between reduced visual acuity and VECP in volunteers with normal corrected visual acuity and in patients suffering from inherited macular degeneration or from age related macular degeneration (ARMD). Flash evoked VECP was not affected by the visual acuity in the cases of refractive error and in ARMD patients but was reduced in amplitude and delayed in implicit time in the patients suffering from inherited macular degeneration. The VECP elicited by pattern reversal checkerboard (PVECP) was not affected by the quality of the visual image in volunteers with uncorrected refractive error when checks of 60' or larger were used but were considerably reduced in size and prolonged in implicit time for checks smaller than 15'. In both groups of patients suffering from macular dysfunction, pattern reversal VECP was very subnormal and was characterized by prolonged implicit time compared to values expected from their visual acuity. These findings indicate that the PVECP does not directly correlate with visual acuity but rather with foveal function. Therefore, we suggest that recordings of PVECP can be used to differentiate between refractive error and macular disorders as causing reduction in visual acuity when other clinical signs are missing or not available. PMID- 11475367 TI - Normal test scores in the Farnsworth-Munsell 100 hue test. AB - One hundred and sixty persons aged from 10 to 69 years (106 women, 54 men) with healthy eyes were studied with the Farnsworth-Munsell 100 hue (FM100) test. The mean of the results in the total scores and in the individual box scores in the right and left eye were calculated. The total score was also separately calculated in women and men. The test was administered under the illumination of Macbeth Easel lamp, 1000 lux, and the right eye was tested first. The results were calculated in six different age groups, 10-19 years, 20-29 years, etc. The mean of the total scores in the right eye varied from 7.44+/-2.46 (SD) to 10.07+/ 2.03 in different age groups and in the left eye from 7.56+/-2.36 to 10.16+/ 2.68. The scores changed significantly with the age: the correlation between the age and the test scores by linear regression gave significant results, in the right eye (R = 0.308, P = 0.0001), and in the left eye (R = 0.246, P = 0.0021). The present study with the normal error scores in the FM100 test and its individual boxes in persons aged 10-69 years gives clinicians working with colour vision defects a possibility to estimate the normality or abnormality of the results in their patients. PMID- 11475366 TI - Multifocal ERG and full-field ERG in patients on long-term vigabatrin medication. AB - PURPOSE: To examine the retinal function with different electrophysiological methods in twelve Swedish patients on long-term treatment (2-10 years) with the anti-epileptic drug vigabatrin. METHODS: Ophthalmological examination of twelve consecutive patients included testing of visual acuity, fundus inspection and fundus photography, kinetic perimetry, full-field ERG and multifocal ERG. RESULTS: All patients had a visual acuity of 0.7 or better. Fundus inspection revealed no pathology except in one patient who had a pallor of the optic disc. All patients had a normal appearance of the macula. The result of kinetic perimetry was normal in five patients while seven patients had a concentric defect of the visual field. The 30 Hz flicker cone b-wave amplitude in the full field ERG was abnormal in all of the seven patients with a visual field defect. None of the patients with normal visual fields had a reduction of the 30 Hz flicker cone b-wave amplitude. Six of the twelve patients had a reduced multifocal ERG response but without any correlation with visual field defect. CONCLUSION: Long-term treatment with vigabatrin seems to selectively reduce retinal cone function. The visual field defects in patients taking vigabatrin correlate with pathology in the full-field ERG (reduction of the cone b-wave amplitude). The results from this study indicate that electroretinography can be used for monitoring patients taking vigabatrin in a more objective manner than with visual field testing. PMID- 11475368 TI - Telecommunications. Static from states. PMID- 11475369 TI - International business. Foreign profits. PMID- 11475370 TI - Outsourcing. Screening ED calls. PMID- 11475371 TI - Child life specialists. An extra comfort. PMID- 11475372 TI - Benchmarking. Guidelines on kids. PMID- 11475373 TI - Language barriers. Plain speaking. PMID- 11475374 TI - The difficult patients live. PMID- 11475375 TI - Health care's most wired. The big payback. PMID- 11475376 TI - Circle of Life Award. AB - For the three winners of this year's Circle of Life Award--Beth Israel Medical Center, New York City; St. Joseph's Manor, Trumbull, Conn.; and Hospice of the North Shore, Evanston, Ill.--life and death are points on a continuous thread. The award is created by the American Hospital Association to recognize innovations in end-of-life care. PMID- 11475377 TI - Facts & fixes. HRET's research agenda complements the AHA's advocacy program. PMID- 11475378 TI - HIV and AIDS--United States, 1981-2000. AB - The acquired immunodeficiency syndrome (AIDS) epidemic has had a substantial impact on the health and economy of many nations. Since the first AIDS cases were reported in the United States in June 1981, the number of cases and deaths among persons with AIDS increased rapidly during the 1980s followed by substantial declines in new cases and deaths in the late 1990s. This report describes the changes in the characteristics of persons with AIDS since 1981. The greatest impact of the epidemic is among men who have sex with men (MSM) and among racial/ethnic minorities, with increases in the number of cases among women and of cases attributed to heterosexual transmission. The number of persons living with AIDS has increased as deaths have declined. Controlling the epidemic requires sustained prevention programs in all of these affected communities, particularly programs targeting MSM, women, and injection drug users. PMID- 11475379 TI - The global HIV and AIDS epidemic, 2001. AB - Human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) are pandemic and pose one of the greatest challenges to global public health. As a bloodborne and sexually transmitted infection, HIV has variable patterns of transmission and impact among world regions and has disproportionately affected disadvantaged or marginalized persons such as commercial sex workers, injection drug users, men who have sex with men (MSM), and persons living in poverty. HIV infection has caused approximately 20 million deaths; an estimated 36 million persons are infected (Figure 1). On the basis of data from the Joint United Nations Program on AIDS (UNAIDS) and other sources, this report summarizes epidemiologic trends, highlights several HIV and AIDS prevention milestones, and describes some prevention activities for the coming decade. PMID- 11475380 TI - HIV incidence among young men who have sex with men--seven U.S. cities, 1994 2000. AB - Twenty years after the first report on human immunodeficiency virus (HIV) infection in the United States, studies of sexually transmitted diseases (STDs) and sexual behaviors suggest a resurgent HIV epidemic among men who have sex with men (MSM). However, few recent studies have measured HIV incidence in this population. To determine HIV incidence among young MSM, CDC analyzed data from the Young Men's Survey (YMS), a study that found a high prevalence of HIV and associated risks among MSM aged 15-22 years sampled in seven U.S. cities. This report confirms high HIV incidence among these young men. PMID- 11475381 TI - The 20th year of AIDS: a time to re-energize prevention. AB - Since the first acquired immunodeficiency syndrome (AIDS) cases were reported in 1981, human immunodeficiency virus (HIV) has caused approximately 22 million deaths worldwide. In the United States, approximately 400,000 persons have died, and approximately one million have been infected. However, numerous persons have avoided infection through prevention efforts, and many lives have been prolonged through advances in treatment. PMID- 11475382 TI - Dynamic analysis of liver allocation policies. AB - Comparisons of alternative liver allocation policies often begin by assuming that patients are either urgent or nonurgent, ignoring the process by which patients become urgent in the first place. This article employs a simulation model to study how patients' health changes between listing and transplant as a function of the rationing rule and the ratio of liver demand to supply. Compared to a first-come first-served queue or random assignment, a "sickest-first" policy results in worse patient outcomes when the demand-to-supply ratio is high. A substantial portion of this differential may be attributed to the fact that under the sickest-first rule, many patients are listed in a nonurgent state but transplanted only once they have reached the sickest patient category. The sickest-first rule is equitable, however, in that patients placed on the waiting list in the sickest category are not disadvantaged relative to patients listed in healthier states. PMID- 11475383 TI - In search of tools to aid logical thinking and communicating about medical decision making. AB - To have real-time impact on medical decision making, decision analysts need a wide variety of tools to aid logical thinking and communication. Decision models provide a formal framework to integrate evidence and values, but they are commonly perceived as complex and difficult to understand by those unfamiliar with the methods, especially in the context of clinical decision making. The theory of constraints, introduced by Eliyahu Goldratt in the business world, provides a set of tools for logical thinking and communication that could potentially be useful in medical decision making. The author used the concept of a conflict resolution diagram to analyze the decision to perform carotid endarterectomy prior to coronary artery bypass grafting in a patient with both symptomatic coronary and asymptomatic carotid artery disease. The method enabled clinicians to visualize and analyze the issues, identify and discuss the underlying assumptions, search for the best available evidence, and use the evidence to make a well-founded decision. The method also facilitated communication among those involved in the care of the patient. Techniques from fields other than decision analysis can potentially expand the repertoire of tools available to support medical decision making and to facilitate communication in decision consults. PMID- 11475384 TI - Revising a priority list based on cost-effectiveness: the role of the prominence effect and distorted utility judgments. AB - BACKGROUND: People sometimes object to the results of cost-effectiveness analysis when the analysis produces a ranking of options based on both cost and benefit. We suggest 2 new reasons for these objections: the prominence effect, in which people attend mostly to a more prominent attrbute (benefit as opposed to cost), and distortion of utility judgments. METHOD: We simulated the production of a cost-effectiveness ranking list in 3 experiments using questionnaires on the World Wide Web. Subjects rated the utility of 16 health benefits using either rating scale or person trade-off elicitation methods. In some experiments, subjects were asked to rate the utility of the health benefits with attention also to the cost of achieving the benefits. In all experiments, at the end, subjects were shown a priority list based on their own utility judgments and were asked whether they wanted to move any of the health benefits up or down the list. RESULTS: In all experiments, subjects wanted to give higher priority to treatments with higher benefit, even when they also had higher cost. They thus wanted to give less weight to high cost (which would, by itself, lead to lower ranking) and more weight to benefit than the weight implied by their own prior judgments. The desire for revision was reduced when subjects made their utility judgments after indicating whether the utility was above or below the midpoint of the scale (a manipulation previously found to reduce distortion). CONCLUSION: The desire to change cost-effectiveness rankings is in part a preference reversal phenomenon that occurs because people attend mainly to the benefit of health interventions as opposed to cost, when they examine the ranking. People should be wary of tinkering with priority lists by examining the lists themselves. PMID- 11475385 TI - An off-the-shelf help list: a comprehensive catalog of preference scores from published cost-utility analyses. AB - PURPOSE: The Panel on Cost-Effectiveness in Health and Medicine recommends an organized collection of preference measure values for health states that can be used in costutility analyses (CUAs). The authors sought to construct a catalog of preference scores from published CUAs, organize the catalog by clinical categories, and identify methods of preference score assessment. METHOD: The authors systematically searched Medline and other databases to identify original CUAs published through 1997. Information was abstracted on the health state descriptions, corresponding preference scores, method of preference score elicitation, and the source of the estimate. RESULTS: Two hundred twenty-eight CUAs were appraised. The authors found 949 health states and corresponding preference scores. Most frequently, health states pertained to the circulatory system (21.7%), health states were valued by experts (35.8%), and values were derived through community-based preference scores (23.5%). CONCLUSION: A catalog of preference scores for health states can be constructed. The catalog (http://www.hsph.harvard.edu/organizations/hcra/cuadatabase/ intro.html) may provide a useful reference tool for producers and consumers of CUAs but also underscores the methodologic variation and inconsistencies present in the field. PMID- 11475386 TI - Stability of patients' preferences for chemotherapy: the impact of experience. AB - BACKGROUND: Studies have shown that utilities for a particular treatment, elicited by means of a hypothetical treatment scenario, may remain stable within the same patients when examined before, during, and after experiencing that treatment (within-group stability). However, other studies have found that utilities for a particular health state may differ between patient groups who are and who are not experiencing the particular health state (between-group differences). OBJECTIVE: The authors evaluated this apparent contradiction in the case of adjuvant chemotherapy for breast cancer. A related purpose was to examine whether a chemotherapy scenario adequately reflects the patients' own experiences with chemotherapy. METHOD: Forty-three patients with early-stage breast cancer evaluated their actually experienced health state and a chemotherapy scenario before, during, and after undergoing adjuvant chemotherapy (chemotherapy group). A control group of 51 patients for whom chemotherapy was not part of the treatment plan was interviewed at similar points in time. Utilities were elicited by means of a visual analog scale (VAS), a chained time trade-off (TTO), and a chained standard gamble (SG). RESULTS: The utilities for the chemotherapy scenario remained relatively stable over time in the 2 patient groups. Furthermore, the chemotherapy scenario was evaluated more positively by patients in the chemotherapy group than by control patients (e.g., utilities before chemotherapy: VAS 0.69 vs. 0.50, TTO 0.88 vs. 0.50, SG 0.92 vs. 0.58, all Ps < 0.01). Finally, patients in the chemotherapy group evaluated their actually experienced health states during chemotherapy higher than the chemotherapy scenario that was assessed at the same time (VAS 0.79 vs. 0.69, TTO 0.93 vs. 0.87, SG 0.97 vs. 0.96, all Ps < 0.05). CONCLUSIONS: Both within-group stability and between-group differences were found. A possible explanation for within-group stability may be that the chemotherapy scenario did not fully correspond to the patients' actual experiences with chemotherapy ("noncorresponding description"). Therefore, preferences did not change even when the patients' own clinical health status had changed. The between-group differences may be explained by "anticipated adaptation." Both explanations may work together to explain why utilities remain stable within the same patients but differ between different patient groups. PMID- 11475387 TI - Does cost-effectiveness analysis make a difference? Lessons from Pap smears. Symposium. PMID- 11475388 TI - Rationing versus rationality: observations from outside the United States. AB - This commentary takes up A. David Paltiel's invitation to reflect on how to promote the use of decision analysis and cost-effectiveness analysis in health. From the perspective of a health services researcher outside the U.S. system, I make 3 arguments. First, the unthinking use of the term rationing for all applications of cost-effectiveness analysis distorts research priorities and may jeopardize wider public support. Second, public skepticism about decision and cost-effectiveness analysis (and thus the skepticism of decision makers) is well founded when ethical dimensions of these methods are not considered. We must continue to refine our methods to take account of societal values. Third, the United States may have particular problems in adopting more rational decision making in health care. The dominance of for-profit institutions in the U.S. health care system erodes the social legitimacy on which other systems depend to improve the rationality of health care decision making. PMID- 11475389 TI - Visual analog scales: do they have a role in the measurement of preferences for health states? AB - Visual analog scales (VASs) have long been used as a method of measuring preferences for health outcomes. They are easy and inexpensive to implement, can be administered quickly, and lend themselves to self-completion. Over time, however, disturbing questions have emerged concerning the validity of the VAS approach. This article reviews briefly the history, theory, practice, problems, and advantages of VASs; presents some suggestions to improve the validity of VASs; and recommends a limited but useful role for VASs in the process of measuring preferences for health states. PMID- 11475390 TI - Law and ethics. PMID- 11475391 TI - Structural changes of the retina in retinal vein occlusion--imaging and quantification with optical coherence tomography. AB - BACKGROUND AND OBJECTIVE: To evaluate the potential of optical coherence tomography (OCT) for imaging and quantifying structural changes in the retinal architecture following venous occlusive disease. PATIENTS AND METHODS: We studied 20 patients with retinal venous occlusive disease: 5 patients suffered from branch retinal vein occlusion (BRVO), and 15 from central retinal vein occlusion (CRVO). Patients with CRVO were studied within 72 hours after the first onset of symptoms. Patients were examined by OCT after a complete ophthalmologic examination including fluorescein angiography. A standard set of linear scans through the center of fixation and individual scans over areas of special interest were performed. Scans were qualitatively evaluated and quantitative measurements were performed on single A-scans at 5 locations of linear scans. Measurements were taken in the center of fixation, at the foveal rim, and at the edge of the scan. We measured total retinal thickness, thickness of cystoid spaces, and thickness of the highly reflective outer band. RESULTS: OCT produced detailed images of retinal thickening, intra- and subretinal hemorrhage, intra- and subretinal fluid accumulation, and formation of intraretinal cystoid spaces. Quantitative evaluation revealed a mean central retinal thickness of 274 +/- 181 microm (72-760 microm) in patients with central retinal vein occlusion. Visual acuity did not correlate with central or extrafoveal retinal thickness. CONCLUSION: Optical coherence tomography is a useful imaging technique for studying changes of retinal architecture following venous occlusive disease. Even minor changes like localized subretinal fluid accumulation or beginning epiretinal membrane formation can be visualized in detail. In contrast to other diseases like diabetic maculopathy, quantitative assessment of macular thickness does not correlate to visual acuity. Nevertheless, because of detailed imaging, OCT might become a useful tool in the follow-up and treatment of individual patients. PMID- 11475392 TI - Suprachoroidal hemorrhage as a complication of vitrectomy. AB - OBJECTIVE: To study the risk factors, management, and end results of suprachoroidal hemorrhage that occur during or after vitrectomy. MATERIALS AND METHODS: This retrospective study involves patients suffering from this complication either during the curing process or immediately after vitrectomy. Preoperative risk factors, operative management, postoperative picture, and end results are reported. RESULTS: During the study period, surgery was performed on 3342 patients with primary vitrectomy. Complications were experienced by 4 patients: 2 occurred near the end of vitrectomy, and 2 in the first postoperative day. The 4 patients were myopic more than 7 diopters, 2 were pseudophakic and 2 were aphakic. Cryopexy was used to treat the retinal breaks in the operative cases. Perfluorophenanthrene was used as a postoperative tamponade in one operative case. Reoperation was done in the 4 patients 3 to 8 weeks after surgery. Silicone oil was used as a prolonged tamponade in all cases. Hypotony persisted in operative cases. All the patients had vision more than 20/400 at the end of follow-up (6 to 24 months). CONCLUSION: The risk factors for suprachoroidal hemorrhage were old age, high myopia, aphakia or pseudophakia, retinal detachment, and scleral buckle. Postoperative suprachoroidal hemorrhage has a better prognosis than the operative type. Perfluorophenanthrene "vitreon" as an operative and postoperative tamponade has a beneficial effect in keeping the retina attached and in preventing pooling of blood under the macula. PMID- 11475393 TI - Subconjunctival anesthesia in contact diode laser cyclophotocoagulation. AB - BACKGROUND: Retro- or peribulbar anesthesia are the standard procedures for cyclodestructive surgery. Because these methods of anesthesia may further compromise optic nerve function, especially in advanced glaucoma, subconjunctival anesthesia was evaluated as an alternative procedure in contact diode laser cyclophotocoagulation (CPC). PATIENTS AND METHODS: A prospective study concerning diode laser CPC in advanced glaucoma was started using subconjunctival anesthesia with 2% mepivacaine. Complications and pain during CPC (5 point rating scale) and on the first postoperative day were recorded. Only the first CPC in every patient was included for evaluation. RESULTS: Included in the study were 120 eyes of 120 patients. During CPC, 82.5% of patients experienced no pain, 11.7% mild pain, and 5.8% moderate pain. No instances of pain or excess motion occurred that required peribulbar anesthesia or discontinuation of the procedure. CONCLUSION: Most of the patients experienced no pain during diode laser CPC using subconjunctival anesthesia. Therefore, risks and side effects of retro- or peribulbar anesthesia can be successfully avoided by this simple modification. PMID- 11475394 TI - Laser therapy in the management of choroidal breast tumor metastases. AB - PURPOSE: To evaluate the potential of laser photocoagulation as a method of treating small metastatic lesions of breast carcinoma in the choroid. METHODS: The 10 eyes of 7 patients were treated by Krypton red or argon green laser applications for small choroidal breast carcinoma metastasis with serous detachment of the retina. Before treatment 5 eyes had visual acuity of finger counting and 5 eyes had visual acuity of 6/15 or better. The treatment was repeated once in all eyes, except 1 eye in which it was repeated 3 times. RESULTS: In all eyes, one to two weeks after the treatment the tumor shrunk, the subretinal serous detachment absorbed, the retina flattened, and the visual acuity improved to 6/6-6/21. No reduction in vision was seen after the second treatment until the end of the follow-up period. In 1 eye, new tumors located elsewhere were again twice successfully treated. Patients reported a significant improvement of their quality of vision, as judged subjectively. CONCLUSIONS: Laser treatment is a feasible, easy, rapid, and effective therapy for small choroidal breast carcinoma. PMID- 11475395 TI - Evaluation of glutaraldehyde and povidone iodine for sterilization of wide-field contact vitrectomy lenses. AB - BACKGROUND AND OBJECTIVE: Wide-field vitrectomy contact lenses are currently sterilized with ethylene oxide gas, and other lenses with autoclaving. To maintain a large inventory or possibly run the risk of loss of lens quality with repeated autoclaving, glutaraldehyde 2% and povidone iodine 5% solution were evaluated as possible sterilizing agents. MATERIALS AND METHODS: Ethylene oxide presterilized lenses were contaminated with known concentrations (10(5) organisms/mL) of bacteria (S. epidemidis, P. aeruginosa, B. subtilis), and fungi (A. flavus, C. albicans) for 5 minutes. The test lenses were treated with glutaraldehyde or povidone iodine for 5, 10, 30, 60, and 120 minutes, and controls with sterilized water for a similar duration. Following treatment, both test and control lenses were sampled with sterile cotton swabs. The swabs were cultured for bacteria (tryptone soya broth 48 hours), and fungi (Saubourd's dextrose broth 5 days). RESULTS: The culture was negative for both glutaraldehyde and povidone iodine-treated lenses against all organisms at all time points except B subtilis, which needed 120 minutes treatment. CONCLUSION: Two hours contact time with glutaraldehyde 2% or providone iodine 5% can sterilize vitrectomy contact lenses against common bacteria and fungi without affecting lens quality. PMID- 11475396 TI - Tectonic epikeratoplasty: a surgical procedure for corneal melting. AB - BACKGROUND AND OBJECTIVE: Tectonic epikeratoplasty (TEK) is a method in which a corneal button, preserved in glycerin is used as a seal over corneal perforation. The graft is sutured to the recipient sclera, upon the sick melted cornea, with silk sutures, after 360 degree peritomy of the conjunctiva. The graft is left in place for a few weeks, and by that time the cornea is completely healed. We describe our experience in TEK as a method of treatment for selected cases of melted cornea or descemetocele. METHODS AND PATIENTS: We review 12 TEK operations done in our department over the period of 1992 to 1997, and describe the outcome in each case. RESULTS: The 12 TEK operations included 9 eyes of 9 patients. The patients, 7 males and 2 females, suffered from ocular surface disturbances including indifference to pain syndrome, Steven-Johnson syndrome, dry eye, relapsing herpetic keratitis, post-traumatic corneal thinning, and local anesthetic abuse. Six of the 9 eyes had leaking perforated corneal ulcer, and the remaining 3 had an imminent perforation caused by descemetocele or melting. All the grafts but one fell off the eyes between 10 and 21 days following surgery. In most of the cases the leakage ceased, and a scar sealing the perforation site was observed. One eye had gone through this procedure 3 times, and another eye had gone through it twice. In only 3 cases did the procedure fail to eliminate leakage, and another TEK or urgent penetrating keratoplasty (PKP) was done. CONCLUSIONS: TEK is an available method for saving the integrity of the globe when PKP is not possible. Although it increases corneal vascularity, PKP can be done later under optimal conditions. PMID- 11475397 TI - Internal limiting membrane removal for traumatic macular holes. AB - OBJECTIVE: To evaluate a new surgical technique for traumatic macular holes and to provide epidemiological information for such holes. MATERIALS AND METHODS: Vitrectomy with internal limiting membrane (ILM) removal but without adjuvant use on 17 consecutive eyes. RESULTS: The hole closed in 100% of eyes. Vision improved > or = 2 Snellen lines in 16 eyes (94%). The macula showed additional trauma related damage in 10 eyes (59%). No permanent complication related to ILM removal was seen. Among 4440 eyes with contusion trauma in the United States Eye Injury Registry, the risk of macular hole formation is 9 times higher in eyes closed than with open globe injury (1.4% versus 0.15%). CONCLUSIONS: Without introducing special risks, removal of the macular ILM appears to be a highly successful surgical option in improving vision in eyes with traumatic macular holes. The majority of eyes benefit from ILM removal, even when additional traumatic macular pathology is present. PMID- 11475398 TI - Toxicity of the photosensitizer NPe6 following intravitreal injection. AB - BACKGROUND AND OBJECTIVE: To determine the retinal toxicity of mono-L-aspartyl chlorin e6 (NPe6) following intravitreal injection. METHODS: Twelve Dutch-belted rabbits divided into 5 experimental groups (n=2 each) were injected intravitreally with 6.25, 12.5, 25, 50, or 100 microg of NPe6; one control group (n=2) was injected with intravitreal normal saline. One eye in each rabbit was sutured shut to test the effect of light exposure. Fundus photography and electroretinograms were performed before treatment and 2 days, 1 week, and 2 weeks after injection. Animals were euthanized and the eyes enucleated for histopathologic analysis. RESULTS: After 1 week, 4 uncovered eyes given 50 and 100 microg had central retinal vein occlusion and varying degrees of retinal hemorrhage. RPE proliferation was seen in the covered eyes given 50 or 100 microg. Electroretinograms revealed absent retinal response at 100 microg and mild toxicity at 50 microg, but no change from normal at doses of < or = 25 microg of NPe6. CONCLUSIONS: Intravitreal doses of < or = 25 microg NPe6 caused little or no apparent toxicity; however, toxicity was significant at doses of 50 microg and 100 microg. PMID- 11475399 TI - Acute bilateral postoperative endophthalmitis in a HLA-DQ5-positive patient: possible susceptibility to infections. AB - An unusual case of bilateral acute endophthalmitis in association with the HLA DQ5 histocompatibility antigen is reported. A 35-year-old woman with high myopia and cataracts in both eyes underwent phacoemulsification procedures performed in separate sessions using different viscoelastic substances and following the same strict prophylactic measures. After each surgical procedure, she developed S. epidermidis acute endophthalmitis in the right eye and Propionibacterium acnes acute endophthalmitis in the left eye; both were successfully treated with capsular bag irrigation and intracameral vancomycin (1 mg/0.1 mL). She tested positive for the HLA-DQ5 (DQ1), DQ2 antigen. The question is raised as to whether the HLA-DQ5 histocompatibility antigen may be a predisposing factor for both staphylococcal and P. Acnes acute endophthalmitis. A multicenter prospective study is proposed to identify this HLA antigen in all patients scheduled for cataract surgery who have a history of postoperative endophthalmitis in the fellow eye. PMID- 11475400 TI - Endogenous Pseudallescheria boydii endophthalmitis in a patient with ring enhancing brain lesions. AB - A 46-year-old man, status post liver transplantation and taking immunosuppressive medications, was admitted after suffering a generalized seizure. Magnetic resonance imaging of the brain revealed two ring-enhancing lesions and treatment was begun for presumed toxoplasmic encephalitis. He was already receiving amphotericin B for a skin lesion suspected to be caused by candidiasis. One day after the seizure, he complained of photophobia in the left eye. Intraocular inflammation and a small infiltrate in the macula were seen. Vision deteriorated over the next three days. Vitreous tap and injection of amphotericin B and vancomycin were performed, but the intraocular inflammation continued to increase. On day 15, a vitrectomy was performed. The vitreous specimen ultimately grew Pseudallescheria boydii. The patient died on hospital day 30 from complications of the brain abscesses. Pseudallescheria boydii should be considered in the differential diagnosis of endophthalmitis, especially in patients with immunosuppression, serious medical disease, or ring-enhancing brain lesions. PMID- 11475401 TI - Valsalva retinopathy-like hemorrhage associated with combined trabeculotomy trabeculectomy in a patient with developmental glaucoma. AB - This case describes valsalva retinopathy-like hemorrhage in a patient with developmental glaucoma who underwent primary combined trabeculotomy trabeculectomy. A 12-year-old boy with developmental glaucoma in both eyes developed shallow anterior chamber following combined trabeculotomy trabeculectomy in his right eye. He underwent air injection for reformation of the anterior chamber under general anesthesia. Following reformation of the anterior chamber on the 5th postoperative day, examination of the patient revealed valsalva retinopathy-like hemorrhage. The patient was treated with betamethasone and cyclopentolate in the postoperative period, and was followed by complete absorption of the retinal hemorrhages with retention of normal visual acuity. Valsalva retinopathy-like hemorrhage may occur with valsalva maneuver during general anesthesia. PMID- 11475402 TI - Cataract surgery with primary posterior chamber intraocular lens implantation in nanophthalmos. AB - A 56-year-old Asian woman with nanophthalmos and cataract in the right eye underwent extracapsular cataract extraction with primary posterior chamber intraocular lens implantation of +31.0 D. Intraoperatively, there was severe vitreous upthrust; however, there was no vitreous loss and the lens could be placed in the posterior chamber. The patient's postoperative course was uneventful except for a moderate degree of coagulum over the lens. Her best corrected visual acuity was 20/80. The patient required YAG capsulotomy one year later. Her intraocular pressure was under control without any medication. With appropriate precautionary measures, primary posterior chamber intraocular lens implantation can be performed in nanophthalmic eyes with a favorable visual outcome. PMID- 11475403 TI - Severe penetrating ocular injury from ninja stars in two children. AB - The authors describe two cases of penetrating ocular trauma in children resulting from ninja stars. In the first case, despite a scleral laceration, loss of iris tissue, and a vitreous hemorrhage, the child had a good result with a final best corrected visual acuity of 20/20. Unfortunately, the child in the second case did not fare as well. In this case, the child suffered a large corneal laceration and traumatic cataract. He ultimately required a penetrating keratoplasty, and he is currently being treated for amblyopia, strabismus, and elevated intraocular pressures. His best corrected visual acuity is 20/70. PMID- 11475404 TI - Rhinosporidiosis and peripheral keratitis. AB - Report of a case of peripheral keratitis caused by Rhinosporidium seeberi. The patient was seen in a referral practice. Corneal scraping was performed on a middle-aged female patient presenting with peripheral keratitis and progressive nasal obstruction that revealed spores suggestive of rhinosporidiosis. The patient was started on topical amphotericin B 0.15% eye drops. Ear, nose, and throat (ENT) examination showed presence of a polypoid lesion in the left nostril for which a polypectomy was performed. Histopathological examination confirmed rhinosporidiosis. Complete resolution of the keratitis was observed. Topical amphotericin B is an effective drug in the management of this condition. Keratitis secondary to rhinosporidial infection has not been described although occasional patients with limbal and scleral involvement have been reported. Corneal scraping was effective in helping us make a tentative diagnosis. PMID- 11475405 TI - Pterygium surgery with mitomycin-C: ten-year results. AB - A review of 870 cases of primary and recurrent pterygia done between 1988 and 1998 by the merest sclera conjunctival flap technique with the single application of 0.1 cc of 0.4 mg/mL of mitomycin-C to the subconjunctival space is reported. There were 3 recurrences (0.35%) and no vision threatening complications. The adverse effects were mild and easily resolved with topical treatment. PMID- 11475406 TI - "Luggage-tag" suture fixation of partially dislocated intraocular lenses. AB - There are many techniques for sulcus-suturing dislocated or partially dislocated intraocular lenses. Many involve passing suture or a knot around the end of the haptic before securing the suture to the sclera. However, in some patients, the tip of the dislocated haptic cannot be visualized. We describe a suturing method using vitrectomy techniques that was used in two patients when the end of the haptic could not be visualized. The technique involves passing an untied loop of double-armed suture under the haptic so that it emerges upward between the haptic and the optic. The loop of suture is regrasped from above the haptic and externalized. The free ends are passed through the loop in a manner analogous to a luggage tag, and the knot is secured. This technique may be useful in patients where the tip of the haptic cannot be visualized, or in patients requiring minimal manipulation of the intraocular lens. PMID- 11475407 TI - Optic nerve metastasis from oat cell lung adenocarcinoma: regression after radiotherapy. PMID- 11475408 TI - Biological timing and the clock metaphor: oscillatory and hourglass mechanisms. AB - Living organisms have developed a multitude of timing mechanisms--"biological clocks." Their mechanisms are based on either oscillations (oscillatory clocks) or unidirectional processes (hourglass clocks). Oscillatory clocks comprise circatidal, circalunidian, circadian, circalunar, and circannual oscillations- which keep time with environmental periodicities--as well as ultradian oscillations, ovarian cycles, and oscillations in development and in the brain, which keep time with biological timescales. These clocks mainly determine time points at specific phases of their oscillations. Hourglass clocks are predominantly found in development and aging and also in the brain. They determine time intervals (duration). More complex timing systems combine oscillatory and hourglass mechanisms, such as the case for cell cycle, sleep initiation, or brain clocks, whereas others combine external and internal periodicities (photoperiodism, seasonal reproduction). A definition of a biological clock may be derived from its control of functions external to its own processes and its use in determining temporal order (sequences of events) or durations. Biological and chemical oscillators are characterized by positive and negative feedback (or feedforward) mechanisms. During evolution, living organisms made use of the many existing oscillations for signal transmission, movement, and pump mechanisms, as well as for clocks. Some clocks, such as the circadian clock, that time with environmental periodicities are usually compensated (stabilized) against temperature, whereas other clocks, such as the cell cycle, that keep time with an organismic timescale are not compensated. This difference may be related to the predominance of negative feedback in the first class of clocks and a predominance of positive feedback (autocatalytic amplification) in the second class. The present knowledge of a compensated clock (the circadian oscillator) and an uncompensated clock (the cell cycle), as well as relevant models, are briefly re viewed. Hourglass clocks are based on linear or exponential unidirectional processes that trigger events mainly in the course of development and aging. An important hourglass mechanism within the aging process is the limitation of cell division capacity by the length of telomeres. The mechanism of this clock is briefly reviewed. In all clock mechanisms, thresholds at which "dependent variables" are triggered play an important role. PMID- 11475409 TI - Multiple oscillators in the suprachiasmatic nucleus. AB - The suprachiasmatic nucleus (SCN) of the hypothalamus is the site of the pacemaker that controls circadian rhythms of a variety of physiological functions. Data strongly indicate the majority of the SCN neurons express self sustaining oscillations that can be detected as rhythms in the spontaneous firing of individual neurons. The period of single SCN neurons in a dissociated cell culture is dispersed in a wide range (from 20h to 28h in rats), but that of the locomotor rhythm is close to 24h, suggesting individual oscillators are coupled to generate an averaged circadian period in the nucleus. Electrical coupling via gap junctions, glial regulation, calcium spikes, ephaptic interactions. extracellular ion flux, and diffusible substances have been discussed as possible mechanisms that mediate the interneuronal rhythm synchrony. Recently, GABA (gamma aminobutyric acid), a major neurotransmitter in the SCN, was reported to regulate cellular communication and to synchronize rhythms through GABA(A) receptors. At present, subsequent intracellular processes that are able to reset the genetic loop of oscillations are unknown. There may be diverse mechanisms for integrating the multiple circadian oscillators in the SCN. This article reviews the knowledge about the various circadian oscillations intrinsic to the SCN, with particular focus on the intercellular signaling of coupled oscillators. PMID- 11475410 TI - Casein kinase I: another cog in the circadian clockworks. AB - Multiple components of the circadian central clock are phosphoproteins, and it has become increasingly clear that posttranslational modification is an important regulator of circadian rhythm in diverse organisms, from dinoflagellates to humans. Genetic studies in Drosophila have identified double-time (dbt), a serine/threonine protein kinase that is highly homologous to human casein kinase I epsilon (CKIepsilon), as the first kinase linked to behavioral rhythms. Identification of a missense mutation in CKIepsilon as the tau mutation in the Syrian hamster places CKIepsilon within the core clock machinery in mammals. Most recently, identification of a phosphorylation site mutant of hPER2 in a family with an inherited circadian rhythm abnormality strongly suggests that PER2 is a physiologically relevant substrate of CKI. Phosphorylation may regulate multiple properties of clock proteins, including stability and intracellular localization. PMID- 11475411 TI - Temporal pattern of LH secretion: regulation by multiple ultradian oscillators versus a single circadian oscillator. AB - The possibility that the 24h rhythm output is the composite expression of ultradian oscillators of varying periodicities was examined by assessing the effect of external continuously or pulsed (20-minute) Gonadotropin-releasing hormone (GnRH) infusions on in vitro luteinizing hormone (LH) release patterns from female mouse pituitaries during 38h study spans. Applying stepwise analyses (spectral, cosine fit, best-fit curve, and peak detection analyses) revealed the waveform shape of LH release output patterns over time is composed of several ultradian oscillations of different periods. The results further substantiated previous observations indicating the pituitary functions as an autonomous clock. The GnRH oscillator functions as a pulse generator and amplitude regulator, but it is not the oscillator that drives the ultradian LH release rhythms. At different stages of the estrus cycle, the effect of GnRH on the expression of ultradian periodicities varies, resulting in the modification of their amplitudes but not their periods. The functional output from the system of ultradian oscillators may superimpose a "circadian or infradian phenotype" on the observed secretion pattern. An "amplitude control" hypothesis is proposed: The temporal pattern of LH release is governed by several oscillators that function in conjunction with one another and are regulated by an amplitude-controlled mechanism. Simulated models show that such a mechanism results in better adaptive response to environmental requirements than does a single circadian oscillator. PMID- 11475412 TI - Phase-shifting effects of dusklike and dawnlike light pulses on the circadian activity rhythms of Syrian hamsters. AB - This study tested whether light pulses with a dusklike offset or a dawnlike onset caused phase shifts of different sizes in the circadian wheel-running activity of Syrian hamsters, Mesocricetus auratus. Six experiments were conducted, each with 30 hamsters; the hamsters received first one type of pulse and then the other type a few weeks later, allowing a paired comparison. The six experiments represented the combination of two maximum light intensities (150 and 250 lux) and three zeitgeber times (ZTs) at which the pulses were given (ZT13.5, ZT14.5, and ZT20). Pulses were 30 minutes long, a relatively short duration to minimize circadian time effects. Aschoff's type II method of measuring phase shifts was used. In none of the six experiments did a two-tailed paired t test detect a significant difference in the size of phase shifts caused by dusklike versus dawnlike pulses. A three-way analysis of variance (ANOVA) on the combined data from all six experiments (with pulse type, pulse intensity, and ZT as factors) also failed to detect a significant effect of pulse type. Statistical power was calculated and found to be reasonably good. These negative results are in line with those of a previous study in which a different methodology was used. PMID- 11475413 TI - Photoperiodic induction of ovarian maturation in crayfish Procambarus clarkii is mediated by extraretinal photoreception. AB - The current study was carried out to investigate whether the photoperiodic induction of ovarian maturation in crayfish is based on a photosensitive rhythm related to extraretinal photoreceptors. To test this, two batches of 61 juvenile crayfish Procambarus clarkii consisting of (1) intact organisms and (2) animals lacking retina and lamina were exposed to 24h light-dark cycles of different photoperiodic schedules based on a night-break protocol for 3 months. Both batches of crayfish showed the greatest ovarian maturation (size, color, degree and size of oocytes) when the light pulse interrupted the scotophase at 21:00 and 05:00, showing a bimodal photoinducible rhythm. Results of the current study indicate that crayfish ovarian maturation depends on a photoinducible rhythm with two possible states that is related to the circadian clock of crayfish. This phenomenon is mediated by extraretinal photoreceptors. Results are interpreted in the light of models of external coincidence. PMID- 11475414 TI - Effect of melatonin on 24h changes in plasma protein levels during the preclinical phase of Freund's adjuvant arthritis in rats. AB - The 24h rhythms in plasma protein concentration were examined in rats on the third day after injection of Freund's complete adjuvant or adjuvant's vehicle, performed 3h after light on. In rats treated with adjuvant's vehicle, peak values of albumin and gamma globulin occurred during the nocturnal activity span (P < .02 and P < .0001, respectively), while those of alpha-1, alpha-2, and beta globulins were found late during the rest span (P < .002, P < .0001, and P < .0004, respectively). Freund's adjuvant administration abolished temporal changes in plasma albumin and beta globulin levels. It also decreased the amplitude of daily changes in alpha-1 and alpha-2 globulin (P < .05) and diminished mean values of alpha-2 globulin (P < .01). Pretreatment of rats with melatonin (30 microg daily) for 11 days, 11h after light on, counteracted mycobacterial adjuvant-induced suppression of the 24h rhythms in albumin and alpha-1, alpha-2, and beta globulins. The results further support the existence of preventive properties of a pharmacological dose of melatonin in situations in which a lost of circadian rhythmicity is expected. PMID- 11475415 TI - Influence of circadian time and age on glomerular angiotensin II receptors in normotensive Sprague-Dawley and transgenic hypertensive TGR(mREN2)27 rats. AB - In male heterozygous transgenic hypertensive rats, TGR(mREN2)27 (TGR), exhibiting an inverse blood pressure profile and in normotensive Sprague-Dawley (SPRD) controls, the density and affinity of angiotensin II receptors were determined at six circadian times in glomeruli of animals 11 weeks old kept under light-dark 12h:12 (LD 12:12) conditions. Angiotensin II receptors were also studied in rats 18-20 weeks old of both strains at 2h after light onset. As a measure of renal excretory functions, diuresis, creatinine, and protein excretion were monitored using metabolic cages. The expression of angiotensin II receptor mRNA was determined in renal arteries 2h-4h after light onset. The following results were obtained: (1) Renal excretory functions showed significant daily variation, with higher excretion rates in the dark span in both TGR and SPRD rats. (2) No circadian phase dependency was found in the glomerular angiotensin II receptors in both rat strains. However, receptor density was significantly lower in TGR than in SPRD rats. In both strains, receptor number increased with aging. (3) In renal arteries, the angiotensin II receptor mRNA of the main receptor subtype AT1A was neither strain nor age dependent, AT1B- and AT2-receptor mRNAs were significantly lower in TGR than SPRD rats. In conclusion, the results demonstrate that the overactive renin-angiotensin system in TGR rats led to a down-regulation of glomerular angiotensin II receptors that was not accompanied by a down regulation of the mRNA of the dominant AT1A- receptor subtype. Circadian short term variations in blood pressure in both TGR and SPRD rats are not reflected by daily variation in angiotensin II receptor density of renal glomeruli or by variation in receptor expression in renal vascular tissue. PMID- 11475416 TI - Cardiovascular regulation in TGR(mREN2)27 rats: 24h variation in plasma catecholamines, angiotensin peptides, and telemetric heart rate variability. AB - Dysfunction of the sympathetic nervous system might play an important role in disturbed 24h blood pressure regulation in transgenic hypertensive TGR (mREN2)27 (TGR) rats. Our study was performed to determine possible differences in activity of the sympathetic nervous system in TGR rats in comparison to their normotensive Sprague-Dawley (SPRD) controls; we measured plasma catecholamine and angiotensin concentrations throughout 24h under synchronized light-dark 12h:12H (LD 12:12) conditions. In the TGR rat strain, rhythms of plasma catecholamines were blunted, and the concentrations were significantly decreased. In addition, TGR rats showed increased plasma angiotensin I and II concentrations without any significant rhythm. An impaired autonomic regulation was confirmed by monitoring heart rate variability in TGR rats. Data showed that the TGR rat strain is characterized by a reduction in plasma catecholamines and an increase in angiotensin peptides. At present, it is not clear whether the reduction in catecholamines represents a decrease in sympathetic tone mediated by baroreflex activation or an increased catecholamine turnover induced by elevated angiotensin II. However, the blunted, but normally phased, rhythms in plasma catecholamines in TGR rats make it unlikely that the sympathetic nervous system is mainly responsible for the inverse circadian blood pressure rhythm in the transgenic strain. PMID- 11475417 TI - Circadian rhythm of double (rate-pressure) product in healthy normotensive young subjects. AB - The double product (DP), systolic blood pressure multiplied by heart rate, is a surrogate measure of myocardial oxygen demand and cardiac workload used increasingly today in medicine. The double product is more strongly correlated with left ventricular mass than the daily blood pressure mean. The purpose of this study was to describe the normative circadian pattern of the double product in healthy normotensive young adults. We studied 125 men and 75 women, 23.0+/-3.3 (mean +/- SD) years of age, without medical history of hypertension and 24h ambulatory systolic/diastolic blood pressure mean consistently below 135/85 mm Hg. Subjects underwent ambulatory blood pressure monitoring at 30-minute intervals for 48 consecutive hours once each season of the year, yielding 930 protocol-correct blood pressure and heart rate time series. Subjects maintained their usual routine of diurnal activity and nocturnal sleep and avoided use of over-the-counter and other medication. Circadian rhythmicity in the double product was established by population multiple-component analysis. The double product rose rapidly from the lowest value, attained 3h before awaking from sleep at night, to a markedly elevated level at the commencement of morning activity. The double product was highest in the afternoon, roughly 7h after the commencement of diurnal activity. In both men and women, the shape of the high amplitude circadian rhythm in the double product was best described by a complex model composed of three cosine curves having periods of 24h, 12h, and 6h. The 24h mean in the double product of 8092.51+/-42.76 (mean +/- SD) in men was significantly lower than that of 8353.17+/-37.48 in women (P < .001). The circadian double amplitude of the rhythm was statistically significantly greater (P < .001) in men (50% of the 24h mean) than women (44% of the 24h mean). The double product did not differ between seasons in women, but it did in men (P = .017) due to reduced heart rate in summer. The circadian pattern of large amplitude in the double product and its gender differences must be taken into account when using this variable to assess cardiac workload, risk of left ventricular hypertrophy, and efficiency of antihypertensive therapy. PMID- 11475418 TI - Gender differences in diurnal variations of subjective activation and mood. AB - This article evaluates the influence of gender on diurnal and postlunch period variations in subjective activation and mood. This topic is not often addressed in the literature; particularly, little attention has been paid to how biological rhythms might bias research results. We studied 40 university student volunteers (20 men, 20 women) aged 18 to 23 years old (X = 20.23, SD = 1.03); they responded to questions on eight unipolar visual analog scales every hour from 08:00 to 21:00. Gender differences were observed in both diurnal and postlunch variations for scales of positive activation (alertness, vigor); sleepiness, however, was only sensitive to diurnal variation, and weariness was sensitive only to a postlunch effect. Women displayed a morning-type pattern, with their optimal moment (11:00) coming 2h earlier than for men, and their activation ratings ranged more widely. The only mood scale that showed differences related to gender was that of happiness, for which women had a higher diurnal mean, a diurnal peak 2h earlier, and a less-intense postlunch effect. Endogenous control of rhythmic pattern appears to be less intense in women, probably due to the coexistence of circamensual rhythmicity, although environmental or sociocultural influences may play a modulating role. Chronopsychological gender differences in affective states should be studied further given the implication they have for the prevention and treatment of mood disorders. PMID- 11475419 TI - Day-night variation in aggressive behavior among psychiatric inpatients. AB - Self-directed aggressive behaviors of human beings show a 24h pattern. The aim of this study was to evaluate if violence of psychiatric inpatients against one another and hospital staff varies over the 24h. The clock time occurrence of 334 episodes of assault behaviors by 119 psychiatric inpatients (78 males and 41 females, mean age 34.8+/-11.3 years) committed during a 5-year span in the psychiatric unit of the university-based hospital of Ferrara, Italy, was evaluated. The clock time of each event was categorized by hour during the 24h and into one of four 6h intervals for analysis of temporal variation by cosinor and chi2 tests, respectively. A significant 24h variation, characterized by an early afternoon peak, was detected irrespective of gender and number (single vs. repeated) of episodes committed. Changes during the 24h in ward activity, patient contact, and endogenous circadian rhythms are likely to contribute to the observed 24h pattern, although further study is needed to confirm our findings and to define causal factors. PMID- 11475420 TI - Actigraphic sleep-wake patterns and urinary 6-sulfatoxymelatonin excretion in patients with Alzheimer's disease. AB - Recent studies suggest melatonin, due to its antioxidant and free-radical scavenging actions, may play a role in the neuroprotection against amyloid, which is implicated in the pathogenesis of Alzheimer's disease (AD). In this study, we determined urinary 6-sulfatoxymelatonin (aMT6s) excretion together with actigraphic sleep-wake patterns of untreated male patients with AD who lived at home. Results were compared with those obtained from normal age-matched elderly and normal young male subjects. Similar measurements were also performed in another group of patients with AD who were treated with a cholinesterase inhibitor (Donepezil, Aricept). Total 24h aMT6s values were significantly reduced in elderly controls (19.9h +/- 5.2 microg/ 24h), in those with untreated AD (12.7 +/- 4.4 microg/24h), and in patients treated for AD (12.4 +/- 4.4 microg/24h) compared with normal young men (32.8 +/- 3.1 microg/24h). A day-night difference in aMT6s was evident in all young controls, in 50% of elderly controls, in only 20% of patients with untreated AD, and in 67% of those with AD receiving Aricept. Sleep quality (expressed as sleep efficiency, wake time, and long undisturbed sleep duration) was better in young and elderly controls compared with the two groups of patients with AD. There was no significant correlation between aMT6s values or sleep patterns and the severity of cognitive impairment in patients with AD. Taken together, these data suggest that disrupted sleep, decreased melatonin production, and partial lack of day-night difference in melatonin secretion were observed equally in normal elderly and in patients with AD. Our results do not permit drawing any conclusion as to whether changes in urinary aMT6s excretion is correlated with disturbed sleep in patients with AD. PMID- 11475421 TI - Season of birth and reproductive performance: an analysis of family reconstitutions of 800 women born in The Netherlands at the end of the 19th century. AB - BACKGROUND: Several studies have reported associations between season of birth and reproductive characteristics such as menarcheal age, fecundability, and twinning, but the results are inconsistent with respect to the location of high- and low-risk seasons. To assess whether this disagreement could be due to the use of populations from different geographic areas and time frames instead of different etiologic pathways, we investigated the season-of-birth dependency of a variety of reproductive outcomes within one time- and area-limited population. METHODS: In a historic follow-up study, the reconstituted families of 800 women born between 1873 and 1887 in or near Rotterdam, The Netherlands, were used to determine eight types of reproductive outcome: childlessness, interval to first pregnancy, pregnancy interval, stillbirth, neonatal death, postneonatal death, multiple birth, and gender of offspring. The relation of these outcomes with season of birth was modeled using cosinor functions with periods of 1 year or a half year. Data were analyzed by use of logistic regression or general estimation equations (GEE), dependent on whether outcomes could occur more than once per woman. RESULTS: Peaks in the model-based risks of reproductive failure were found within two small temporal ranges, January 1 to February 11 and July 1 to August 11 for all outcomes except gender. The picture did not change after controlling for known and possible risk factors, including age, offspring's birth cohort, and some social variables. CONCLUSIONS: This study reconfirms the idea that seasonal factors around conception or birth influence later reproductive characteristics. Observing the consistency of the location of high-risk seasons across a variety of outcomes, the explanation of season-of-birth dependency of different reproductive outcomes need not involve multiple etiological pathways. PMID- 11475422 TI - Influence of dietary intake and physical activity on annual rhythm of human blood cholesterol concentrations. AB - Seasonal variation in the plasma total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) have been repeatedly reported, with contradictory results regarding the pattern of seasonal variation of these parameters. Furthermore, it is still not well established whether the variation is due to changes in the nutrition or changes in physical activity depending on the season. The aim of this study was therefore to determine plasma TC and HDL-C in different groups of healthy participants: 19 vegetarians with a constant diet independent of the season, 14 athletes with almost constant physical activity over the year, and 114 controls in the age groups 20-26 years (mean age 24 + 1.5 years) and 40 48 years (mean age 44.3 + 2.1 years). Over 2 years, blood samples were collected every 2-3 months and were analyzed for plasma TC and HDL-C. At all visits, body mass index (BMI) and waist-to-hip ratio (WHR) were calculated, and nutrition and physical activity profiles were obtained. The seasonal model was calculated using object-oriented software for the analysis of longitudinal data in S (OSWALD); multiple regression analysis was used to determine the influence of age, gender, diet, and physical activity on seasonal changes of the lipid parameters. In all groups, we found an annual rhythm of the plasma TC and HDL-C concentrations, which can be mathematically described by a sine curve with a maximum in winter and a minimum in summer. This rhythm was independent of the age, gender, BMI, diet, or physical activity. The observed seasonal differences between the maximum and the minimum were about 5%-10% for TC and about 5%-8% for HDL-C concentration. These differences were greater than the determined circadian (TC 3.5%, HDL-C 4%) and day-to-day changes for TC and HDL-C (coefficient of variation <5% for both). In conclusion, annual rhythm of TC and HDL-C is not primarily induced by seasonal differences in dietary intake or physical activity. Therefore, the annual rhythm in cholesterol levels is most likely determined by endogenous factors or factors directly related to seasonal changes in the environment. PMID- 11475423 TI - Impaired expression of the mPer2 circadian clock gene in the suprachiasmatic nuclei of aging mice. AB - The expression of circadian clock genes was investigated in the suprachiasmatic nuclei (SCN) of young adult and old laboratory mice. Samples were taken at two time points, which corresponded to the expected maximum (circadian time 7 [CT7]) or minimum (CT21) of mPer mRNA expression. Whereas the young mice had a stable and well-synchronized circadian activity/rest cycle, the rhythms of old animals were less stable and were phase advanced. The expression of mPerl mRNA and mPer2 mRNA was rhythmic in both groups, with peak values at CT7. The levels of mClock and mCry1 mRNA were not different depending on the time of day and did not vary with age. In contrast, an age-dependent difference was found in the case of mPer2 (but not mPerl) mRNA expression, with the maximum at CT7 significantly lower in old mice. The decreased expression of mPer2 may be relevant for the observed differences in the overt activity rhythm of aged mice. PMID- 11475424 TI - Chronotoxicology of florfenicol. AB - Sixty 3-month-old homozygote male mice were studied for circadian rhythmicity in the toxicity of florfenicol overdose. Animals were kept under a regimen of 12h light, 12h darkness (12:12 LD) with food and water available ad libitum. The LD50 (median lethal) dose was determined in a preliminary experiment and was administered to groups of 10 mice at six different clock times (hours) after light onset (HALO): 0, 4, 8, 12, 16, and 20 HALO. Cosinor analysis verified a statistically significant (P < .04) circadian rhythm in the toxic effect (mortality) of florfenicol. Mortality was greatest when the drug was injected 4h after the commencement of the activity span (16 HALO) and least when injected 4h after the start of the diurnal rest span (4 HALO). Mortality was 2.5 times greater when drug injection was given at 16 HALO than at 4 HALO. PMID- 11475425 TI - The sources of knowledge in psychogeriatrics. PMID- 11475426 TI - Gender-related cognitive deficits in Alzheimer's disease. AB - Recent studies have identified cognitive deficits in semantic memory and verbal language abilities among women with Alzheimer's disease (AD). Few studies to date have explored gender differences in episodic memory function in AD. The present study compared the performance of men and women diagnosed with AD on a battery of neuropsychological measures. Results indicated the presence of gender-related cognitive deficits on tasks of confrontation naming, expressive word knowledge, and both episodic and semantic memory for women with AD, relative to findings in men. PMID- 11475427 TI - Further evidence of westernization of dementia prevalence in Nagasaki, Japan, and family recognition. AB - The present study examined the prevalence of dementia in the Nagasaki Prefecture. The purposes of our investigation were (a) to study the relationship between aging and the prevalence of dementia and the ratio of Alzheimer's disease (AD) to vascular dementia (VD), (b) to understand the features of early-onset dementia as seen in patients from 60 to 65 years, and (c) to examine the recognition of dementia by family members. The subjects of the study, a total of 4,368, were all 60 years old and over and were residing in the three areas of Nagasaki Prefecture at the time of the investigation, August 1995. We adopted a two-stage design. The first-stage questionnaire that we developed was delivered to subjects, and we selected for the second stage those subjects who met the criteria outlined in the Methods section. The second-stage investigation was an interview by community nurses and psychiatrists. The prevalence of dementia in subjects 60 years and over was 6.2% (men: 5.9%; women: 6.8%). The prevalence increased with age. The AD/VD ratio was 1.4, and was similar to the recent trend in Japan in that the ratio has reversed to resemble the western pattern. In regard to the family members' recognition of illness, the higher the severity of dementia, the higher the recognition ratio of family members became. Only half of these subjects were recognized as having dementia by their family members. In conclusion, the westernization of the AD/VD ratio in Japan was proved. There was little study about family recognition of dementia. In this study, it was remarkable that only half of the subjects were recognized as having dementia by their family members. PMID- 11475428 TI - A revised CES-D measure of depressive symptoms and a DSM-based measure of major depressive episodes in the elderly. AB - This study examines the psychometric properties of two new abbreviated versions of standard measures of depression, a revised eight-item Center for Epidemiological Studies-Depression Scale (CES-D) and a short-form Composite International Diagnostic Interview (short-form CIDI). A sample of 6,133 elders, age 70 years or older, completed both measures as part of the Asset and Health Dynamics Study of the Oldest Old. The revised CES-D had an internal consistency and factor structure comparable to that of prior versions of the CES-D. The sources of discordance between the two measures were examined and the two measures were compared on self-report of four clinical variables: medical illness, physician diagnosis, psychiatric treatment, and antidepressant or tranquilizer use. Both measures were associated with self-report of physician diagnosis and psychiatric treatment. Respondents positive for depression on the CES-D reported higher rates of antidepressant use. Respondents positive on the short-form CIDI only did not report more antidepressant use than nondepressed respondents. PMID- 11475429 TI - Predictors of recovery from major depression among geriatric psychiatry inpatients: the importance of caregivers' beliefs. AB - Caregiver support is an important factor in recovery from depression among older patients. We examined whether caregivers' perceptions regarding patients' ability to control depressive symptoms were related to depression recovery. Depression treatment, demographics, number of depressive symptoms, and health were controlled. The sample comprised 51 geriatric psychiatry inpatients who met DSM IV criteria for major depression and who had a primary caregiver. Depression was assessed at both admission and discharge. Caregivers were asked to indicate whether they believed their patient-relatives could control their depressive symptoms. At discharge, 33 patients (64.7%) were "remitted" and 18 (35.3%) were "nonremitted." Multivariate analyses indicated that receiving electroconvulsive treatment, having fewer depressive symptoms caregivers perceived to be within patient control, and being female predicted depression remission at discharge. This study highlights the important relationship between family dynamics and course of depression. PMID- 11475430 TI - Reliability and validity of the subjective burden scale in family caregivers of elderly relatives with dementia. AB - The reliability and validity of the Subjective Burden Scale (SBS) were investigated in 255 Japanese family caregivers of elderly relatives with dementia. Values of the Cronbach's alpha reliability, split-half reliability (r), and test-retest reliability (r) were .87, .80, and .72, respectively. The SBS score was significantly positively correlated with the degree of mental health, which was assessed with the Japanese version of the 60-item General Health Questionnaire (r = .41). The family caregivers who gave up the caregiving of their relatives showed significantly higher SBS scores than those who did not. The SBS scores for caregivers indicated satisfactory predictive validity both in the degree of their mental health and in the prospects of family caregiving about 6 months later. These findings suggest that the SBS is adequately reliable and valid and can be used to assess the subjective burden of Japanese caregivers. PMID- 11475431 TI - Determination of normative criteria and validation of the SKT for use in Spanish speaking populations. AB - Most of the tests, questionnaires, and neuropsychological batteries for the assessment of dementia have been translated and adapted for use in the Hispanic population without having normative data, which results in a high number of false positives when age and educational level are not considered. The Short Cognitive Performance Test (SKT) is a psychometric instrument evaluating memory and attention deficits that has been developed and standardized in Germany (Erzigkeit, 1989a, 1989b). The objective of this study was to adapt the SKT, to establish normative criteria that take into consideration age and educational level, and to establish its concurrent validity in comparison to other neuropsychological tests: Brief Neuropsychological Evaluation for Spanish Speaking Subjects (Ostrosky-Solis et al., 1994), the Mini-Mental State Examination (Folstein et al., 1975), and the Blessed Functional Scale (Blessed et al., 1968) in 238 neurologically intact subjects and 97 subjects with mild to moderate dementia. The SKT showed adequate sensitivity (80.5%) and specificity (80.3%) in subjects with medium and high educational level; however, the sensitivity and specificity diminished (75% and 56.7%) in subjects with no education or low educational level. The adapted and validated version of the SKT in the Mexican population has been shown to be a psychometric instrument that in subjects with medium educational level can detect cognitive alterations and is able to determine the severity of deterioration; however, in subjects with low educational level and severe dementia, the SKT cannot be usefully administered. PMID- 11475432 TI - Revised memory and behavior problems checklist in Taiwanese patients with Alzheimer's disease. AB - The Revised Memory and Behavior Problems Checklist (RMBPC) is a 24-item caregiver report that measures observable behavioral and memory problems in dementia patients and their caregivers' reaction to these problems. The purpose of the present study was to evaluate the applicability of the RMBPC for use in Taiwanese patients with Alzheimer's disease (AD). The subjects included 76 AD patients (39 men and 37 women, mean age 72.3) and their caregivers (34 men and 42 women, mean age 53.5) who participated in a comprehensive assessment at the Veterans General Hospital-Taipei. The Chinese version of the Cognitive Abilities Screening Instrument was administered to the patients. Their caregivers rated the RMBPC and the short version of the Geriatric Depression Score (GDS). To assess the test retest reliability, 30 caregivers rated a second RMBPC 3 days after the first evaluation. The mean score for the frequency rating on the RMBPC was 32.63 (SD = 12.44, range = 5-61) and the mean reaction score was 10.96 (SD = 11.53). The reaction score was significantly correlated with the GDS score (r = .363, p = .001). The Cronbach's alpha coefficients for frequency and reaction scores were .816 and .895 respectively. The test-retest reliabilities of total frequency and reaction scores were significantly correlated; overall correlations were .89 for frequency (p < .001) and .74 for reaction (p < .001). These findings suggest that the RMBPC be recommended as a reliable tool to assess behavioral and memory disturbance in Taiwanese AD patients. PMID- 11475433 TI - Pharmacological and psychotherapeutic treatment of personality disorders in the elderly. AB - Although estimates as to the incidence of personality disorder in the elderly remain controversial, it is well known that such a disorder is prevalently ego syntonic and capable of interfering with the onset and treatment of other somatic and psychic pathologies, especially in later life, when individuals tend tobe particularly vulnerable. Misdiagnosis or failure to treat these disorders may greatly diminish the quality of life of older adults and their families. The aim of this work is to define the chief psychopharmacological and psychotherapeutic guidelines for treating personality disorders in the elderly. Pharmacologically, patients require treatment with molecules designed to rebalance neurotransmitter system alterations, which underlie the symptomatological picture. As regards psychotherapeutic treatment, although there are only a few validation studies, we believe that dialectical behavior therapy and interpersonal psychotherapy may constitute valid therapeutic approaches that meet both the needs and the individual characteristics of patients affected by personality disorder and those of elderly patients. PMID- 11475434 TI - Modeling decline in Alzheimer's disease. PMID- 11475435 TI - In search of a disease marker: the cytokine profile of primary progressive multiple sclerosis. PMID- 11475436 TI - Intracellular cytokine profile in T-cell subsets of multiple sclerosis patients: different features in primary progressive disease. AB - OBJECTIVE: To evaluate the expression of cytokines in both CD4+ and CD8+ T cells derived from peripheral blood of untreated multiple sclerosis (MS) patients with either relapsing-remitting (RR), secondary progressive (SP) or primary progressive (PP) MS and healthy controls (HC). BACKGROUND: MS is an immune mediated disease and cytokines hove been hypothesized to contribute significantly to disease progression. Compared to the relapse-onset (RR, SP) form of the disease, PPMS patients have different clinical, immunological and pathological features. Surprisingly, the ability of their circulating T cells to produce immunoregulatory cytokines has not been extensively studied so far. METHODS: Seventy-two MS patients (24 RR, 26 SP, 22 PP) and 34 HC were studied. Stimulated peripheral blood derived CD4+ and CD8+ T MS patients express significantly more CD4+ and CD8+ T cells were analyzed for IFN-gamma, IL-2, TNF-alpha, IL-4, IL-10 and IL-13 production. RESULTS: cells producing IFN-gamma compared to HC. Compared to the other forms of the disease, PPMS patients display a significant decrease in CD4+ T cells producing IL-2, IL-13 and TNF-alpha and a significant increase in CD8+ T cells producing IL-4 and IL-10. CONCLUSIONS: The data presented here demonstrate that patients with PPMS express less pro- and more anti-inflammatory cytokine producing T cells compared to the relapse-onset form of the disease, confirming the view on PPMS as a distinct disease entity. PMID- 11475437 TI - Similar pro- and anti-inflammatory cytokine production in the different clinical forms of multiple sclerosis. AB - Cytokines play an important role in the initiation and maintenance of the inflammatory reaction in multiple sclerosis, a chronic inflammatory demyelinating disease of the central nervous system. Magnetic resonance imaging evidence supports clinical divergence between forms of multiple sclerosis with relapses and the primary progressive form without relapses, which shows fewer and smaller inflammatory lesions. With the aim of understanding better the relative role of pro-inflammatory and/or anti-inflammatory cytokines in primary progressive multiple sclerosis in comparison to relapsing forms, we analysed in 65 patients (24 primary progressive, 20 relapsing-remitting and 21 secondary progressive) and 29 healthy controls, the production of cytokines (IFN-gamma, TNF-alpha, IL-6, IL 10 and IL-12) by peripheral blood mononuclear cells after in vitro stimulation. We found a similar percentage of cytokines producing cells between healthy controls and the different clinical forms of multiple sclerosis patients. PMID- 11475438 TI - Analysis of an interferon-gamma gene dinucleotide-repeat polymorphism in Nordic multiple sclerosis patients. AB - The proinflammatory cytokine interferon (IFN)-gamma has been shown to influence the course of multiple sclerosis (MS). The IFN-gamma (IFNG) contains a multiallelic dinucleotide repeat in intron 1. To investigate whether alleles at this locus influence susceptibility to MS, we performed linkage and familial association analyses on 100 sibling pairs from four Nordic countries, and case control association analysis on 220 intermediately disabled sporadic MS patients and 266 controls. To determine the effect of the polymorphism on disease outcome, we compared genotype frequencies in the most and least disabled octiles of a total cohort of 913 cases. We also measured IFN-gamma mRNA levels in unstimulated peripheral blood mononuclear cells from 46 MS patients and 27 controls grouped according to IFNG intron 1 genotype. Both nonparametric linkage analysis and transmission disequilibrium testing of the 100 sibling pairs produced negative results. Genotype frequencies for intermediate-MS patients did not differ significantly from those for controls; nor did genotype frequencies in the benign MS octile differ significantly from those in the severe-MS octle. Comparison of IFN-gamma mRNA levels in genotype-conditioned subgroups revealed no significant differences. Thus, alleles at the IFNG intron 1 dinucleotide repeat appear to affect neither MS susceptibility and severity nor IFN-gamma mRNA expression in vivo. PMID- 11475439 TI - Uric acid levels in patients with multiple sclerosis: analysis in mono- and dizygotic twins. AB - Presence of nitrotyrosine in cells surrounding plaques indicates that peroxynitrite may be the cause of brain lesions in multiple sclerosis. Low levels of uric acid, a natural scavenger of peroxynitrite, were demonstrated in blood of patients with multiple sclerosis in comparison with control individuals. These observations were now extended to 132 sets of twins with one sibling affected by multiple sclerosis. In blood of both mono- and dizygotic twins the uric acid levels were lower in the twin with the disease than in the healthy twin. PMID- 11475440 TI - Ring-enchancement in multiple sclerosis: marker of disease severity. AB - Correlations between conventional MRI measures of disease activity and clinical disability in multiple sclerosis (MS) have been disappointing. Because ring enhancing lesions may reflect a more destructive pathology, we tested their potential association with disease severity. We evaluated active lesions with regard to their enhancement pattern on serial magnetic resonance images in a cohort of 28 patients with relapsing-remitting MS. The percentage of ring enhancing lesions correlated with EDSS, T2 lesion load and duration of disease and predicted the occurrence of relapses during the baseline period of observation as well as after 3 years of follow-up in multiple logistic regression analysis. The findings suggest that the pathological process reflected by ring enhancing lesions may contribute to more severe clinical disease. PMID- 11475441 TI - Clinical, radiological, immunological and pathological findings in inflammatory CNS demyelination--possible markers for an antibody-mediated process. AB - The present report describes immunopathological, radiologcal and serological characteristics of antibody-mediated demyelination in a multiple sclerosis (MS) case with the main findings: (1) immunoglobulin and complement deposits in areas of active demyelination accompanied by massive macrophage activation; (2) ring enhancing lesions in T1-weighted MRI after gadolinium application; (3) high titers of serum anti-myelin antibodies; and (4) signs of macrophage activation in the serum. Plasmapheresis may be a successful treatment for the type of inflammatory demyelination shown in the present case. PMID- 11475442 TI - Double-blind randomized multicenter dose-comparison study of interferon-beta-1a (AVONEX): rationale, design and baseline data. AB - We describe the rationale and design of a double-blind, randomized, multicenter, dose-comparison study of interferon-beta-1a (IFN-beta-1a; AVONEX), in the treatment of relapsing multiple sclerosis (MS). The study is expected to provide quantitative insights on the dose range for optimal clinical benefits in MS. The study involves 802 patients in 10 European countries who have EDSS scores 2.0 5.5, and who have experienced at least two relapses within the 3 years prior to enrolment Patients are randomized to receive once-weekly intromuscular injections of IFN-beta-la 30 or 60 mcg for at least 3 years. The primary endpoint of the study is the effect of IFN-beta-1a therapy on the time to sustained progression of disability. For patients with a baseline EDSS < or = 4.5, sustained progression of disability is defined as a 1 point increase in EDSS from baseline, maintained for 6 months. For patients with baseline EDSS > or = 5, sustained progression of disability is defined as reaching an EDSS > or = 6.0, maintained for 6 months. EDSS scores will be determined every 3 months. A series of prospectively defined secondary and tertiary efficacy endpoints will be examined. Safety will be monitored throughout the study. Magnetic resonance imaging (MRI) with and without gadolinium-enhancement has been performed in at least 358 patients at baseline and repeated annually after enrolment In a subset of these patients, a frequent MRI study is also being performed. PMID- 11475443 TI - Effect of monthly intravenous cyclophosphamide in rapidly deteriorating multiple sclerosis patients resistant to conventional therapy. AB - Fourteen consecutive clinically definite relapsing-remitting multiple sclerosis (MS) patients were treated with monthly intravenous cyclophosphomide (CTX) for 6 months. All had experienced severe dinical deterioration during the 12 months prior to treatment with CTX despite treatment with conventional immunomodulating agents and intravenous methylprednisolone. Treatment with CTX led to improvement and neurologic stability within 6 months which was sustained for at least 18 months after the onset of treatment with CTX. Therapy with CTX was well tolerated. CTX may be of benefit in MS patients who experience rapid clinical worsening and are resistant to conventional therapy. PMID- 11475444 TI - Profiles of nursing home residents with multiple sclerosis using the minimum data set. AB - This paper profiles nursing home residents with multiple sclerosis (MS) at the time of admission, including sociodemographic characteristics, health status measures, and treatments received. Admission assessments from the Minimum Data Set are used to create these profiles of residents with MS. There are 9,013 admission assessments in the MDS for residents with MS between June 22, 1998 and January 17, 2000 analyzed for this study. Residents with MS are distinctly younger at admission than most nursing home residents, averaging 57.98 years of age. Recently admitted residents with MS are more physically dependent than other nursing home residents and tend to have limited range of motion and loss of voluntary movement About one in three newly admitted residents with MS had some degree of impaired cognitive function. Over one third of residents with MS were depressed at admission, yet only 11.7% of recently admitted residents with MS were evaluated by a licensed mental health specialist This prompts concem about the psychosocial well-being of MS residents in nursing homes. PMID- 11475445 TI - Self-administered Expanded Disability Status Scale with functional system scores correlates well with a physician-administered test. AB - BACKGROUND: Patient-administered measures are needed to assess disability cost effectively in large epidemiological studies. SETTING: An outpatient clinic in a large multiple sclerosis center. METHODS: A self-administered EDSS questionnaire was developed (EDSS-S). Consecutive patients with clinically definite multiple sclerosis completed the EDSS-S (n=95). During the same visit, a physician completed an EDSS (EDSS-P). Scores below 4.0 were determined using functional system (FS) scores. Scores above 4.0 were calculated by two methods, using gait alone and using gait and functional system scores combined. RESULTS: EDSS-P scores ranged from 0-9.5 (mean 5.1, median 5.0, 78% female, age 17-78, mean age 45). Mean EDSS-P, EDSS-S and intraclass correlation coefficients of agreement were: EDSS using ambulation alone (4.6, 5.1, 0.89), EDSS using ambulation and FS scores (4.6, 5.3, 0.87), bowel/bladder FS scores (1.6, 1.7, 0.79), pyramidal FS scores (2.1, 2.4, 0.67), sensory FS scores (1.6, 2.1, 0.60), cerebellar FS scores (1.1, 1.6, 0.55), brainstem FS scores (0.5, 1.2, 0.45), vision FS scores (1.9, 1.3, 0.38), cerebral FS scores (0.6, 2.3, 0.27). CONCLUSIONS: Very good correlation was seen between patient and physician scores for EDSS and the bowel/bladder FS score. Four other FS scores correlated moderately. In general, patients scored themselves more disabled than physicians. PMID- 11475446 TI - Nutrition and physical activity interventions to reduce cardiovascular disease risk in health care settings: a quantitative review with a focus on women. AB - The authors conducted a quantitative literature review of the impact of 32 diet and physical activity (PA) interventions delivered in health care settings on cardiovascular disease risk factors. Intervention effects were relatively modest but statistically significant for PA, body mass index or weight, dietary fat, blood pressure, and total and low-density lipoprotein serum cholesterol. Intervention effects were generally larger for samples with a mean age >50 years and for studies with <6 months follow-up. Type of comparison group, type of intervention, and use of a behavior theory did not have a consistent impact on intervention effects. Few studies focused on persons of color, although the results from these studies are promising. PMID- 11475447 TI - Fetal malformations and folate metabolism: review of recent evidence. AB - Although a reduction in incidence of neural tube defects is unequivocally linked to adequate folate status, evidence is also mounting associating folate with other fetal malformations. The emerging discoveries about single nucleotide polymorphisms have given new insight into folate biochemistry, enabling more precise understanding of how genetic variations influence folate-dependent pathways in embryogenesis. Findings suggest that folate status may be partly under genetic control, and may involve a "cocktail effect" resulting from interactions among genes, nutrients, and enzymes. Despite major laboratory advances, much of the human evidence comes from observational studies, and questions linger that cannot be definitively answered without randomized clinical trials. PMID- 11475448 TI - Family and child-care provider influences on preschool children's fruit, juice, and vegetable consumption. AB - Children's intakes of fruit, juice, and vegetables (FJV) do not meet the recommended minimum of five daily servings, placing them at increased risk for development of cancer and other diseases. Because children's food preferences and practices are initiated early in life (e.g., 2-5 years of age), early dietary intervention programs may have immediate nutritional benefit, as well as reduce chronic disease risk when learned healthful habits and preferences are carried into adulthood. Families and child-care settings are important social environments within which food-related behaviors among young children are developed. FJV preferences, the primary predictor of FJV consumption in children, are influenced by availability, variety, and repeated exposure. Caregivers (parents and child-care providers) can influence children's eating practices by controlling availability and accessibility of foods, meal structure, food modeling, food socialization practices, and food-related parenting style. Much remains to be learned about how these influences and practices affect the development of FJV preferences and consumption early in life. PMID- 11475449 TI - Efficacy of garlic supplementation in lowering serum cholesterol levels. AB - Previous studies using garlic have found alterations on a number of cardiovascular disease (CVD) risk factors including blood pressure, plasma viscosity, platelet activity, and serum lipid levels. The latest clinical research suggests that consumption of garlic powder does not play a significant role in lowering plasma lipid levels when in conjunction with a low-fat, low cholesterol diet. Additional well-controlled, long-term studies that explore dosage and preparation type are necessary to confirm the efficacy of garlic in lowering cholesterol levels and to fully understand garlic's potential role in CVD. PMID- 11475450 TI - Does lowering plasma homocysteine reduce vascular disease risk? AB - A recent study indicates that lowering homocysteine with folic acid and vitamin B6 significantly reduces the incidence of abnormal exercise electrocardiography tests and thus decreases the risk of atherosclerotic disease. Questions about study design and sample size, however, suggest that this conclusion may be premature. PMID- 11475452 TI - Outcomes following cheilectomy and interpositional arthroplasty in hallux rigidus. AB - Surgical management of hallux rigidus remains controversial. Arthrodesis is considered the gold standard. However, many patients are reluctant to undergo fusion. This paper reviews two commonly used procedures that are reasonable alternatives. A retrospective review of 19 patients (24 feet) with grade 2 osteoarthritis and 11 patients (11 feet) with grade 3 osteoarthritis was performed. The patients with grade 2 osteoarthritis were managed with a cheilectomy and the patients with grade 3 osteoarthritis with an interpositional arthroplasty. All patients were individually assessed with a subjective questionnaire, physical exam, AOFAS hallux scale, SF-36 and pedobarographic analysis. Cheilectomy patients (51.9 years) were younger than interpositional arthroplasty (59 years). Follow up between the interpositional arthroplasties (2.0 years) and cheilectomies (2.1 years) were comparable. Postoperative motion, visual analogue pain scale and SF-36 scores were comparable between groups. Cheilectomies had a higher mean AOFAS score (77.3) than interpositional arthroplasties (71.6). Weakness of the great toe was reported in 72.7% of interpositional arthroplasty patients compared to only 16.7% of patients with a cheilectomy. Patient satisfaction was 87.5% in cheilectomies and 72.7% in interpositional arthroplasties. Pedobarographic analysis demonstrated a decreased load under the great toe with increased weight transfer to the lesser metatarsal heads in all patients. The weight transfer to the lesser metatarsal heads was greatest in patients with interpositional arthroplasty. Management of moderate hallux rigidus with a cheilectomy and phalangeal osteotomy is a reliable method of relieving pain and improving function. Management of severe osteoarthritis of the joint with an interpositional arthroplasty should be considered a salvage procedure with less reliable results. PMID- 11475453 TI - Pediatric calcaneal fractures. AB - Calcaneal fractures in pediatric patients are seen infrequently. We retrospectively reviewed 22 skeletally immature patients with 23 fractures of the calcaneus before their distal tibial physis had fused. Eighteen (78%) of these fractures were intraarticular and five (22%) were extraarticular. Nine patients were followed for an average of 4.4 years. Of these nine fractures, 8 were treated non-operatively and one was treated with open reduction and internal fixation. A modification of the American Orthopaedic Foot and Ankle Society scoring system, which focuses on residual complaints of pain, work or sports restrictions, ability to walk on different surfaces and gait abnormalities, was employed to assess their outcome. Seven of the nine patients were free of pain, had unrestricted foot function, and possessed no apparent gait abnormalities. Two patients had activity related and cold weather related pain. We believe that the excellent prognosis for both intraarticular and extraarticular fractures, seen in this study is multi-fold. First, the immature talus and calcaneus have a superior capacity to remodel. Second, pediatric calcaneal fractures are usually consequence of low energy trauma. Lastly, a favorable intraarticular fracture pattern, unique to the skeletally immature calcaneus may exist, rendering a good prognosis. PMID- 11475454 TI - Healing of the Achilles tendon: an experimental study. AB - Biomechanical properties of healing ruptures in the Achilles tendon of rabbits were examined after two, four, eight and 12 weeks. Treatment modalities were (n7): a) suture, b) fibrin-glue, c) non-surgical treatment. All animals received a functional aftertreatment consisting of a special orthotic support and free ambulation. For biomechanical testing a fixation-technique was applied that guaranteed intratendinous rupture. After two weeks, the tendons treated with fibrin glue showed better results (stiffness, maximum force to rupture, tensile stress to rupture) compared with the non-surgically treated group. The results for sutured tendons were in between those for the other groups. After four weeks, the results for sutured and for glued tendons were nearly equal and slightly better than the tendons in the non-surgical group. Late results revealed comparable biomechanical properties among all treatment groups and control tendons, suggesting our conclusion that non-surgical treatment is equal to repair using sutures or using fibrin glue as measured by stiffness and tensile stress. PMID- 11475455 TI - Syme ankle disarticulation: a simplified technique. PMID- 11475456 TI - Force patterns of heel strike and toe off on different heel heights in normal walking. AB - This study investigated the changes of force patterns of the heel strike and toe off phases at different heel heights during normal walking. Ten healthy female college students wore running shoes, flat leather shoes and high heeled shoes while walking on a Kistler force platform at their self-comfortable paces. It was found that the high heeled shoes and the leather shoes generated significantly greater vertical impact forces and anterior-posterior forces in the toe off phase than those in the heel strike phase. Accumulated impulses did not show significant increase while the heel heights increased and total support time while wearing the high heeled shoes was significantly longer than while wearing the running shoes. PMID- 11475457 TI - The measurement of the medial longitudinal arch in children. AB - Both feet of two hundred and seventy two children aged between five years six months and ten years and eleven months were studied using a footprint technique called the arch index (Al), and the vertical height of the navicular (NH) as non invasive techniques of objective measures of the medial longitudinal arch (MLA). In addition to age the study investigated the influence of gender, limb dominance, and body weight. The study found the existence of a relationship between the two measures of the MLA. There was no significant difference in NH measures between males and females and body weight did not affect the NH. The NH changed with age, suggesting it provides a useful, easily obtained clinical measure. The Al measures were slightly more reliable than the NH but showed less change with age. PMID- 11475458 TI - 'Ostoesynthesis' of a symptomatic bipartite medial cuneiform. AB - Bipartition of the medial cuneiform is uncommon and often not recognized on plain radiographs. It is usually asymptomatic and rarely, if ever, requires surgery. Injury to the synchondrosis of a bipartite medial cuneiform is rare and has, to our knowledge, been reported only once. We describe such a case with chronic disabling midfoot pain after remote trauma. PMID- 11475459 TI - Pneumatic bracing and total contact casting have equivocal effects on plantar pressure relief. AB - The purpose was to examine and compare plantar pressures produced in healthy subjects while wearing a running shoe (RS), total contact cast (TCC) and 'customized' pneumatic pre-fabricated walking brace (PWB). A repeated measures design was used to compare the plantar pressures recorded for three footwear types (RS, TCC, PWB) in two body regions (forefoot, heel). Nine healthy subjects walked at a self-selected walking pace on a motorized treadmill while wearing the RS, TCC and PWB (ordered randomization). Following a five-minute acclimatization period on the treadmill with each footwear device, plantar pressures were recorded from 84 constant gait speed and step length steps using the Pedar system of in-shoe array of capacitive sensors embedded in an insert. Mean spatially averaged peak plantar pressures were recorded for the metatarsal heads and heel region for each footwear device worn by each subject. A two-way analysis of variance with repeated measures and post-hoc Tukey tests analysed the data with a significance level of p=.05. The main effects of footwear (p=.005) and body region (p=.000), and interaction effect (body region x footwear device) (p=.000) were significant. Unloading of the forefoot was 63.72% and 58.77% for the TCC and PWB, respectively, whereas loading under the heel was increased 37.09% and 34.11% for the same two devices, respectively. Patients who develop neuropathic plantar ulcers in the forefoot region, but not in the heel region, may benefit from a reduction in plantar pressures by using either the TCC or a 'customized' PWB. An alternative footwear device still needs to be found for those patients with heel ulceration. PMID- 11475460 TI - Benign calcaneal bone cyst and pathologic fracture--surgical treatment with injectable calcium-phosphate bone cement (Norian): a case report. AB - Solitary calcaneal bone cysts are uncommon. Usually they measure 1/3 to 1/2 of the calcaneal length. Symptomatic calcaneal bone cysts are generally treated with open debridement and autologous bone grafting. We report a case of a patient with a displaced intra-articular calcaneal fracture who presents with a large benign calcaneal bone cyst. This patient was treated with debridement and filling of defect with injectable calcium-phosphate bone cement (Norian) and open reduction and internal fixation of the calcaneal fracture. PMID- 11475461 TI - Congenital crossover fifth toe correction with soft tissue release and cutaneous Z-plasty. AB - Congenital overriding of the fifth toe causes symptoms in approximately half of all patients. The results of three patients treated with a simpler procedure involving metatarsal phalangeal capsular release dorsally and medially, oblique lengthening of the extensor tendon, and Z-plasty of the skin is reported. At an average of 33 month follow-up, no recurrence is noted with this simple procedure. PMID- 11475462 TI - Autologous chondrocyte transplantation in osteochondral lesions of the ankle joint. AB - The aim of this study was to assess the repair of osteochondral defects of the ankle joint with hyaline cartilage. For this purpose we have been using a technique of autologous chondrocyte transplantation for osteochondral defects of the talus for the last two years. Until the method described in the paper, treatment methods proposed for the repair of cartilaginous defects have not been histologically effective in restoring the hyaline cartilage sheath, and in all cases the neoformation of cartilage was of a fibrocartilaginous nature with varying cellular characteristics. Clinical and histological results obtained using this surgical technique have confirmed its validity. Furthermore, neither subjective nor objective complications have been reported. Less pain and better articular function have also been observed. According to the AOFAS score, an improvement from an average score of 32/100 points pre-op. to 91/100 points at 24 months of follow up was obtained. Laboratory data have confirmed the presence of reconstructed cartilage with chondrocytes and expression of collagen II, characteristic of hyaline cartilage. PMID- 11475463 TI - Re: "Comparison of results of retrocalcaneal decompression for retrocalcaneal bursitis and insertional Achilles tendinosis with calcific spur" by Dr. Anthony Watson et. al. PMID- 11475464 TI - A cavovarus foot is a predisposing factor for rather than a result of peroneus longus tendinopathy. PMID- 11475465 TI - Pharmacokinetic software for the health sciences: choosing the right package for teaching purposes. AB - Computer assisted learning has an important role in the teaching of pharmacokinetics to health sciences students because it transfers the emphasis from the purely mathematical domain to an 'experiential' domain in which graphical and symbolic representations of actions and their consequences form the major focus for learning. Basic pharmacokinetic concepts can be taught by experimenting with the interplay between dose and dosage interval with drug absorption (e.g. absorption rate, bioavailability), drug distribution (e.g. volume of distribution, protein binding) and drug elimination (e.g. clearance) on drug concentrations using library ('canned') pharmacokinetic models. Such 'what if' approaches are found in calculator-simulators such as PharmaCalc, Practical Pharmacokinetics and PK Solutions. Others such as SAAM II, ModelMaker, and Stella represent the 'systems dynamics' genre, which requires the user to conceptualise a problem and formulate the model on-screen using symbols, icons, and directional arrows. The choice of software should be determined by the aims of the subject/course, the experience and background of the students in pharmacokinetics, and institutional factors including price and networking capabilities of the package(s). Enhanced learning may result if the computer teaching of pharmacokinetics is supported by tutorials, especially where the techniques are applied to solving problems in which the link with healthcare practices is clearly established. PMID- 11475466 TI - Therapeutic drug monitoring: do the improved outcomes justify the costs? AB - In the 30 years that therapeutic drug monitoring (TDM) has moved from an abstract consideration to a routine intervention, issues remain over justifying the benefits in the light of the ever-increasing competition for budgetary resources. Resolving the issues is constrained by various methodological concerns. These include considerations such as: (i) the changed environment of knowledge and practice during the generation in which TDM has been used and evaluated; (ii) the predominance of studies using system-related rather than patient-centred outcomes; (iii) using a timeframe for analysis that is too short; (iv) a lack of rigour in many of the pharmacoeconomic analyses; and (v) excessive use of a site specific rather than a societal perspective. Current observation suggests that the greatest benefit of TDM accrues from targeted or specialty populations: those with severely decompensated renal function, those at the extremes of age, and those using immunosuppressive, some antineoplastic, some psychotherapeutic and some anticonvulsant drugs. In these situations, safe and humane practice considers TDM a necessity without respect to cost. But for many routine situations with drugs for which TDM has commonly been used in the past, present reliance on the intervention may have become excessive in the light of today's knowledge base of practitioners. PMID- 11475467 TI - Pharmacokinetic studies with esomeprazole, the (S)-isomer of omeprazole. AB - This article reviews the pharmacokinetics of esomeprazole, the (S)-isomer of the proton pump inhibitor (PPI) omeprazole. Esomeprazole is the first single isomer PPI developed for the treatment of patients with acid-related diseases. In vitro experiments in human liver microsomes demonstrated that the formation of the hydroxy and 5-O-desmethyl metabolites of esomeprazole is via cytochrome P450 (CYP) 2C19, whereas that of the sulphone metabolite is via CYP3A4. The formation rate of the hydroxy metabolite from esomeprazole is lower than for (R) omeprazole, but that of the 2 other metabolites is higher, demonstrating stereoselective metabolism. The sum of the intrinsic clearances of all 3 metabo- lites for esomeprazole was one-third of that for (R)-omeprazole, suggesting lower clearance of esomeprazole in vivo. In vivo investigations demonstrated that esomeprazole is chirally stable after administration. Esomeprazole is 97% bound to plasma proteins. In normal (extensive) metabolisers with regard to CYP2C19, esomeprazole is metabolised more slowly than omeprazole, resulting in a higher area under the concentration-time curve (AUC) after administration of the same dose. This is more pronounced after repeated administration rather than after a single dose. In poor metabolisers, the AUC is lower for esomeprazole than for omeprazole, contributing to less overall interindividual variability for esomeprazole than for omeprazole. In general, esomeprazole and omeprazole are subject to the same metabolic transformations. Almost complete recoveries were reported and the ratio between urinary and faecal excretion is about 4:1 for both compounds. The dose-dependent increase in AUC of esomeprazole with repeated administration results from a combination of decreased first-pass elimination and decreased systemic clearance. Patients with gastro-oesophageal reflux disease exhibit a pharmacokinetic pattern similar to that in healthy individuals, whereas elderly individuals exhibited a slightly lower metabolism rate. Patients with a severe deficit in their liver function had a lower rate of metabolism, as would be expected, whereas those with mild to moderate liver disease did not exhibit any alteration in the pharmacokinetics. The pharmacokinetics of esomeprazole in individuals with impaired renal function is unlikely to differ from that in healthy individuals. A slight sex difference in the pharmacokinetics of esomeprazole was demonstrated in that the AUC and peak plasma drug concentration were slightly, but not statistically significantly, higher in females than in males. PMID- 11475468 TI - Clinical pharmacokinetics of inhaled budesonide. AB - The corticosteroid budesonide is a 1:1 racemic mixture of 2 epimers, (22R)- and (22S)-, and is available in 3 different inhaled formulations for the management of asthma: a pressurised metered dose inhaler (pMDI), a dry powder inhaler (DPI) and a solution for nebulised therapy. Inhaled corticosteroids such as budesonide reach the systemic circulation either by direct absorption through the lungs (a route that is much more important than previously recognised) or via gastrointestinal absorption of drug that is inadvertently swallowed. Although the pharmacokinetics of budesonide have been extensively investigated following oral and intravenous administration, relatively few studies have defined the systemic disposition of budesonide after inhalation. Drug deposition in the lungs depends on the inhaler device: 15% of the metered dose of budesonide reached the lung with a pMDI compared with 32% with a breath-actuated DPI. In patients with asthma (n = 38) receiving different doses of budesonide by DPI (Turbuhaler), the pharmacokinetic parameters peak plasma concentration (Cmax) and area under the concentration-time curve (AUC) were dose-dependent after both single dose and repeat dose (3 weeks) administration: time to Cmax (tmax) was short (0.28 to 0.40 hours) and the elimination half-life approximately 3 hours. Both AUC and Cmax were linearly related to budesonide dose. In a small group of healthy male volunteers (n = 9), the pharmacokinetics of budesonide 1,600 microg twice daily via pMDI were assessed on the fifth day of administration. Mean model-independent parameters for (22R)-budesonide were as follows: Cmax 1.8 microg/L, tmax 0.46 hours, elimination half-life 2.3 hours and oral clearance 163 L/h, and there were no enantiomer-specific differences in drug disposition. Budesonide undergoes fatty acid conjugation within the lung, but very limited pharmacokinetic data are available to define the pulmonary absorption characteristics. There is evidence from a population analysis that the pulmonary absorption of budesonide is prolonged and shows wide interindividual variation. Further pharmacokinetic studies are required to define the time-course of budesonide absorption through the lung in specific patient groups, and to investigate the effect of new inhaler devices (especially chlorofluorocarbon-free pMDIs) on the pharmacokinetic profile and systemic drug exposure. PMID- 11475470 TI - Pharmacokinetics of cyclosporin microemulsion in patients with inflammatory bowel disease. AB - OBJECTIVE: To obtain a pharmacokinetic profile of cyclosporin microemulsion formulation in patients with inflammatory bowel disease. PATIENTS AND PARTICIPANTS: 58 consecutive patients (19 women and 39 men), aged 16 to 64 years (mean age 38 years), with a diagnosis of ulcerative colitis (29 patients) or Crohn's disease (29 patients). METHODS: Patients were treated with oral doses of cyclosporin microemulsion ranging from 200 to 400 mg daily. Blood samples were collected after 7 days of treatment; blood was drawn at 0, 0.5, 1, 2, 3, 5, 7 and 12 hours after the morning dose. In 23 patients out of 29 with ulcerative colitis and 23 out of 29 with Crohn's disease, these profiles were repeated immediately before hospital discharge, which took place an average of 18 days after admission. Blood specimens were assayed for cyclosporin immunoreactivity on the day of blood withdrawal by a radioimmunoassay technique. MAIN OUTCOME MEASURES AND RESULTS: In the range of doses employed, the average peak plasma drug concentration (Cmax) and area under the concentration-time curve to 12 hours tended to increase linearly with the dose (from 782.35 to 1,607.98 microg/L and from 3,612 to 7,221 microg x h/L for doses of 200 mg and 400 mg, respectively), whereas the time to Cmax (tmax) and elimination half-life (t 1/2beta) ranged between 78 and 95.2 min and 85.5 and 162 min, respectively, and did not appear to change with the dose. After dose-normalisation by transformation of data into percentage increase over baseline (trough) concentration for each patient, single kinetic parameters for the whole study population (n = 58) could be calculated (Cmax 620% vs baseline. tmax 86.5 min, t 1/2 115 min). Comparison between patients with Crohn's disease and ulcerative colitis showed that the latter had higher Cmax values (702% compared with 543% vs baseline, p < 0.05) whereas tmax and t 1/2beta values overlapped. CONCLUSIONS: The pharmacokinetic parameters of cyclosporin microemulsion in patients with inflammatory bowel disease are broadly similar to those previously measured in healthy volunteers and in other disorders requiring cyclosporin treatment. PMID- 11475472 TI - Erythropoietin in tumor therapy. 10-11 November 2001, Berlin, Germany. Abstracts. PMID- 11475469 TI - Pharmacokinetics of antifungal agents in onychomycoses. AB - Onychomycosis is caused by infection by fungi, mainly dermatophytes and nondermatophyte yeasts or moulds; it affects the fingernails and, more frequently, the toenails. Dermatophytes are responsible for about 90 to 95% of fungal infections. Trichophyton rubrum is the most common dermatophyte; Candida albicans is the major nondermatophyte yeast. Although topical therapy of onchomycosis does not lead to systemic adverse effects or interactions with concomitantly taken drugs, it does not provide high cure rates and requires complete compliance from the patient. At present there are 3 oral antifungal medications that are generally used for the short term treatment of onychomycosis: itraconazole, terbinafine and fluconazole. The persistence of these active drugs in nails allows weekly administration, reduced treatment or a pulse regimen. Good clinical and mycological efficacies are obtained with itraconazole 100 to 200 mg daily, terbinafine 250mg daily for 3 months, or fluconazole 150 mg weekly for at least 6 months. Itraconazole is a synthetic triazole with a broad spectrum of action. It is well absorbed when administered orally and can be detected in nails 1 to 2 weeks after the start of therapy. The nail : plasma ratio stabilises at around 1 by week 18 of treatment. Itraconazole is still detectable in nails 27 weeks after stopping administration. Nail concentrations are higher than the minimum inhibitory concentration (MIC) for most dermatophytes and Candida species from the first month of treatment. The elimination half-life of itraconazole from nails is long, ranging from 32 to 147 days. Terbinafine is a synthetic allylamine that is effective against dermatophytes. Terbinafine is well absorbed from the gastrointestinal tract, and the time to reach effective concentrations in nail is 1 to 2 weeks. The half-life is from 24 to 156 days, explaining the observed persistence of terbinafine in nails for longer than 252 days. Fluconazole is a bis-triazole broad spectrum antifungal with high oral bioavailability. The uptake of fluconazole by nail increases with the length of treatment, and nail : plasma ratios are generally 1.5 to 2 at steady state. Fluconazole concentrations exceed the MIC for Candida species soon after the start of treatment. Fluconazole concentrations fall slowly after the drug is stopped, with a half-life of 50 to 87 days, and fluconazole is still detectable in nails 5 months after the end of treatment. All these drugs are potent inhibitors of cytochrome P450 (CYP) enzymes and may increase the plasma concentrations of concomitantly used drugs. Itraconazole inhibits CYP3A4. Fluconazole inhibits CYP3A4, but to a lesser degree than itraconazole, CYP2C9 and CYP2C19. Terbinafine inhibits CYP2D6. PMID- 11475473 TI - Smoking for nicotine. PMID- 11475474 TI - Promoting smoking cessation in the rehabilitation setting. AB - Cigarette smoking is a known risk factor in patients with ischemic and hemorrhagic stroke. Smoking also increases the risk of cardiovascular disease, chronic bronchitis, emphysema, peptic ulcers, and cancer of several organs among middle-aged individuals and the elderly. In the elderly, smoking has also been associated with a general decline in physical functioning as a result of the increased incidence of chronic illnesses. The prevalence of smoking among community-dwelling adults aged 65 to 74 yr has been estimated to be 18% for men and 15% for women. More than 30% of Americans who are hospitalized each year are smokers. Although there are no published studies that have established the prevalence of smoking in a rehabilitation population, these data and our own clinical experience suggest that smoking continues to be a significant health problem for many persons who enter the inpatient rehabilitation setting. Because most hospitals have adopted a smoke-free policy, hospitalization itself may initiate a period of nonsmoking in patients who were smokers at the time of their admission. In addition, some smokers choose to quit smoking after stroke or other medical crisis caused by the health risks associated with cigarette smoking. However, research has also revealed a rather low-smoking cessation rate (30%) among smokers who have had a transient ischemic attack despite the health benefits associated with smoking cessation. Given the significant health risks associated with cigarette smoking, particularly in the elderly and those with cerebrovascular compromise, the effects of smoking on the patient's health should be discussed with the patient during inpatient rehabilitation. Unfortunately, given the current healthcare demands of reducing lengths of hospitalization and the focus on functional outcomes, health promotion issues, such as smoking cessation, nutrition, exercise, may not receive the attention that they deserve. Despite these constraints, we believe that the inpatient rehabilitation setting provides an opportunity for a "teachable moment" to introduce the idea of smoking cessation to the active smoker or to encourage continued smoking cessation and relapse prevention to those patients who have not smoked since their admission to the acute care hospital. If instituted in an effective manner, we believe that there could be significant healthcare benefits in establishing a formal smoking cessation or relapse prevention program in the rehabilitation setting. PMID- 11475475 TI - Use and satisfaction with prosthetic devices among persons with trauma-related amputations: a long-term outcome study. AB - OBJECTIVE: To document and examine the use, satisfaction, and problems with prosthetic devices among persons who suffered a trauma-related lower limb amputation. DESIGN: Abstracted medical records and follow-up interview data were collected for a retrospective cohort of persons with a lower limb trauma-related amputation who received their acute care at the University of Maryland R. Adams Cowley Shock Trauma Center, Baltimore, MD, between 1984 and 1994. Patients with spinal cord injury, traumatic brain injury, or only toe amputations were excluded. RESULTS: There were 146 patients identified. Of those, 9% died during the acute admission and 3.5% died after discharge. Seventy-eight amputees were available for interview (68% response rate). The majority of those interviewed were male (87%), and two-thirds had undergone amputation before age 40 yr. Nearly 95% had a prosthesis and wore it an average of 80 hr (SD = 33) per week. Despite high use, only 43% reported being satisfied with the comfort of their prosthesis. About one-quarter of all users reported problems with wounds, skin irritation, or pain. Traumatic amputees used an average of four prostheses since injury, about one new prosthesis every 2 yr. Statistical analyses revealed that males reported higher prosthetic use (P < 0.01). Higher Injury Severity Score negatively impacted on prosthetic use (P < 0.01). Phantom pain negatively influenced reported satisfaction with the prosthesis (P < 0.03) CONCLUSIONS: Although almost all persons living with trauma-related amputations use prosthetic devices, the majority are not satisfied with prosthetic comfort. Phantom pain and residual limb skin problems are also common afflictions in this population. PMID- 11475476 TI - Relationship between hip muscle imbalance and occurrence of low back pain in collegiate athletes: a prospective study. AB - OBJECTIVE: To assess whether athletes with strength imbalance of the hip musculature would be more likely to require treatment for low back pain (LBP) over the ensuing year. DESIGN: The study population included 163 National Collegiate Athletic Association Division I college athletes (100 males and 63 females) undergoing preparticipation sports physicals. Institutional review board approval was obtained to acquire and analyze hip muscle strength data. A commercially available dynamometer (Chatillon, Lexington, KY) incorporated into a specially designed anchoring station was used for testing the hip extensors and abductors. The maximum force generated for the hip abductors and extensors was used to calculate a percentage difference between the right and left hip extensors and abductors. Treatment of athletes by the athletic trainers for LBP unrelated to blunt trauma over the ensuing year was recorded. RESULTS: Of all athletes, 5 of 63 females and 8 of 100 males required treatment for LBP. Logistic regression analysis indicated that for female athletes, the percentage difference between the right and left hip extensors was predictive of whether treatment for LBP was required over the ensuing year (P = 0.05). There was no significant association noted for the percentage difference between the right and left hip abductors in females and for the percentage difference between both the right and left hip abductors and right and left hip extensors in males requiring treatment for LBP. CONCLUSIONS: These data support our results from our previous cohort study, adding validity to the concept of hip muscle imbalance being associated with LBP occurrence in female athletes. This research further supports the need for the assessment and treatment of hip muscle imbalance in individuals with LBP. PMID- 11475477 TI - Brace modification improves aerobic performance in Charcot-Marie-Tooth disease: a single-subject design. AB - OBJECTIVE: Ankle-foot orthoses (AFOs) can lower energy expenditure in patients with hemiplegia by 10%-13%. Review of the lower motor injury literature reveals insufficient physiologic evidence supporting the use or modification of AFOs in patients with lower motor neuron injury and, specifically, progressive conditions such as Charcot-Marie-Tooth disease. We sought to test the hypothesis that optimal AFOs would improve submaximal aerobic performance and submaximal perceived exertion, while producing no change in maximal aerobic capacity. DESIGN: In an individual with Charcot-Marie-Tooth disease, a single-subject design study was used. An A-B-A design was used, with "A" corresponding to use of the patient's old AFOs and "B" corresponding to the newly prescribed AFOs. The subject underwent treadmill exercise tolerance testing using a modified Balke protocol. Indirect calorimetry was used to measure oxygen consumption per unit time (VO2), and the Borg scale was used to measure perceived exertion. RESULTS: At the same submaximal exercise intensities, VO2, rate-pressure product, and perceived exertion were all reduced when using the modified AFOs. Additionally, these conditions allowed the subject to conduct the treadmill exercise test 20% longer. Maximal VO2 remained constant under all conditions. CONCLUSION: Optimizing the AFO prescription in a patient with Charcot-Marie-Tooth disease can enhance physiologic performance and perceived exertion at submaximal activity levels. Larger controlled trials are necessary to further demonstrate such benefits in patients with progressive neuropathy and other causes of lower motor neuron injury. PMID- 11475478 TI - Physical treatment of Peyronie disease. AB - OBJECTIVE: Peyronie disease is a localized and progressive fibrosis. It is characterized by a plaque in the tunica albuginea, which leads to penile deformity, making sexual intercourse difficult, if not impossible. DESIGN: During a 4-yr period, we treated 35 patients, aged 30-62 yr, in different stages of this disease. We applied ultrasound therapy (0.5 W/cm; 10 min), infrared radiation, and iontophoresis with 8% potassium iodide (0.2 mA; 30 min). The patients were taught to administer therapy by themselves. The patients' diseases were classified into three stages on the basis of subjective symptoms and clinical findings. At the beginning of treatment, 20 patients' diseases were classified as being in the first stage, 13 patients' diseases in the second stage, and 2 patients' diseases in the third stage. RESULTS: By the end of treatment, 10 patients were cured, 17 patients' diseases were classified as being in the first stage, 8 patients' diseases were in the second stage, and there were no patients in the third stage. CONCLUSIONS: The method is simple, safe, painless, and inexpensive. Patients were taught to administer the therapy by themselves. There were no side effects. Functional improvement and the cessation of pain were noted by all the patients. The level of improvement depended on the disease duration, the length of therapy, and the stage of the disease. PMID- 11475479 TI - Predicting models of outcome stratified by age after first stroke rehabilitation in Japan. AB - OBJECTIVE: A multivariate model predicting the function at discharge following inpatient rehabilitation has been previously produced. The aim of this study is to determine predictors of function at discharge for stroke outcome and examine their accuracy of prediction. DESIGN: Four hundred sixty-four stroke patients were enrolled. Sex, the nature of the stroke, age, onset to rehabilitation admission interval and length of rehabilitation hospital stay were obtained from their medical records. Patients were divided into the following five groups according to age: < or = 49, 50-59, 60-69, 70-79, and > or = 80 yr. Disability was assessed on admission and at discharge by the FIM. Stepwise multiple regression analysis was performed in each group. RESULTS: The model for patients aged 60-69 yr was best for accuracy of prediction and explained 76% of variation for discharge FIM total score. The equation: (expected discharge FIM total score) = 111.88 + 0.08 x (the type of stroke) - 0.11 x (age) + 0.81 x (admission FIM total score) - 0.12 x (onset to rehabilitation admission interval), R = 0.87, R2 = 0.76, P < 0.0001. The type of stroke = 1 for cerebral infarction and 0 otherwise. Length of rehabilitation stay is not selected as a predictor. CONCLUSION: The stratification of patients by age is useful to determine predictors of function at discharge for stroke outcome and to improve their accuracy of prediction. PMID- 11475480 TI - Bone mineral density in patients with stroke. AB - OBJECTIVE: Stroke is an acute neurologic dysfunction of vascular origin, characterized by loss of voluntary movement, sensory disturbances, and neurologic findings in the contralateral half of the body. Acute and long-term complications because of immobilization are seen in all organ systems. The aim of this study was to determine any differences between the affected and unaffected sides' bone mineral densities of acute and chronic stroke patients. DESIGN: In this study, we determined the bone mineral densities (BMD) of 30 male patients with acute (0-20 days) and 30 male patients with chronic (6 months or longer) stroke and compared the densities with the normal side. Upper and lower limb BMDs were measured by dual-energy x-ray absorptiometry. Additionally, patients were evaluated for the degree of spasticity, the phases of motor improvement, and the activities of daily living. RESULTS: In acute-phase stroke patients, BMD of the affected side was not significantly different from BMD of the normal side. BMD of both upper limbs of acute stroke patients was not different from normal side upper limb BMD of chronic stroke patients. In both acute and chronic stroke patients, affected and unaffected side lower limb BMDs (femur total BMD scores) were not significantly different. However, in chronic stroke patients, affected side BMD of Ward's region was significantly higher compared with the normal side. There is no correlation between BMD and Brunnstrom phases, Ashworth scales, and the degree of activities of daily living. CONCLUSIONS: Higher BMD of the affected side Ward's region in chronic cerebrovascular accident patients may be related to spasticity and changes in walking pattern, which increase the mechanical stress loading of the Ward's region. PMID- 11475481 TI - Virtual reality in the assessment of selected cognitive function after brain injury. AB - OBJECTIVE: To assess selected cognitive functions of persons with traumatic brain injury using a computer-simulated virtual reality environment. STUDY DESIGN: A computer-simulated virtual kitchen was used to assess the ability of 30 patients with brain injury and 30 volunteers without brain injury to process and sequence information. The overall assessment score was based on the number of correct responses and the time needed to complete daily living tasks. Identical daily living tasks were tested and scored in participants with and without brain injury. Each subject was evaluated twice within 7 to 10 days. A total of 30 tasks were categorized as follows: information processing, problem solving, logical sequencing, and speed of responding. RESULTS: Persons with brain injuries consistently demonstrated a significant decrease in the ability to process information (P = 0.04-0.01), identify logical sequencing (P = 0.04-0.01), and complete the overall assessment (P < 0.01), compared with volunteers without brain injury. The time needed to process tasks, representing speed of cognitive responding, was also significantly different between the two groups (P < 0.01). CONCLUSION: A computer-generated virtual reality environment represents a reproducible tool to assess selected cognitive functions and can be used as a supplement to traditional rehabilitation assessment in persons with acquired brain injury. PMID- 11475482 TI - Trampolines, children, and strokes. AB - Strokes in children related to sports injuries are rare, but pediatric trampoline injuries are dramatically increasing. Minor trauma to the vulnerable extracranial vertebral arteries as they travel superficially through the dorsum of the neck can begin a cascade of events that results in arterial dissection, thrombus formation, and embolization with cerebral infarction. We present the case of an 11-yr-old boy who developed left vertebral artery dissection subsequent to a trampoline injury. PMID- 11475483 TI - Continuous passive motion in the management of heterotopic ossification in a brain injured patient. AB - We report a man admitted to inpatient rehabilitation 6 wk after traumatic brain injury, who presented with bilateral knee heterotopic ossification. In addition to conventional physical therapy, we applied a continuous passive motion device during 4 wk increasing the range of motion of the knees. On the basis of the limited current literature and this case, we suggest that the use of continuous passive motion devices for heterotopic ossification may be effective and safe and should be the subject of further study. PMID- 11475484 TI - Persistent hiccup associated with thoracic epidural injection. AB - Epidural steroid injections are commonly used to treat lumbosacral radicular and discogenic pain. When used in this manner, these agents can cause minor, transient systemic side effects and rarely result in any serious complications. Because adverse reactions are uncommon and transient, epidural injections are considered a safe therapeutic intervention. We describe the first case of persistent hiccups as a consequence of a thoracic epidural steroid injection in a patient with thoracic discogenic pain. PMID- 11475485 TI - Accessory nerve injury as a complication of cervical lymph node biopsy. PMID- 11475486 TI - Increasing resident physician initiation of vocational rehabilitation services. PMID- 11475487 TI - Preliminary observations on reduced utilization in skilled nursing facility rehabilitation. PMID- 11475488 TI - Healthy People 2000: an assessment based on the Health Status Indicators for the United States and each state. AB - The Health Status Indicators (HSIs) were developed as part of the Healthy People 2000 process to facilitate the comparison of health status measures at national, State and local levels. In this report, the number of HSIs where the national target has been attained and the number of HSIs that have improved significantly are enumerated for the United States, the District of Columbia, and each State. Based on data for 1998, the United States had attained targets for 6 of the 17 HSIs with national targets. Twenty-three States had attained targets for 9 or more of the HSIs and 37 States had either attained the target for, or had made significant progress on, at least 12 of the indicators. Substantial progress has therefore been made since the Healthy People 2000 National Health Promotion and Disease Prevention Objectives were published in 1991. PMID- 11475489 TI - Good cop, bad cop: federal prosecution of state-legalized medical marijuana use after United States v. Lopez. AB - The Supreme Court's recent decisions in United States v. Lopez and United States v. Morrison articulate a vision of federalism under which Congress's regulatory authority under the Commerce Clause is severely limited in favor of returning traditional areas of state concern, particularly criminal law enforcement, to local or state control. The Court's decisions in these cases coincide with ballot initiatives legalizing the medical use of marijuana garnering a majority of the vote in California, Arizona, Alaska, Colorado, Nevada, Oregon, Washington, Maine, and Washington D.C. Those who use marijuana for medical purposes under sanction of state law, however, still face the threat of federal prosecution under the Controlled Substances Act. Medical marijuana proponents have traditionally, and unsuccessfully, contested federal prosecution using individual rights arguments under theories of equal protection or substantive due process. This Comment argues that after Lopez and Morrison, the federal government's authority to regulate intrastate use of marijuana for medicinal purposes is not the foregone conclusion it once was. The author suggests that proponents of medical marijuana use should invoke the federalism arguments of Lopez and Morrison and argue for state legislative independence from the federal government on this issue. PMID- 11475490 TI - Infliximab: new preparation. A last resort for Crohn's disease. AB - (1) The licensed indications for infliximab, an immunosuppressive agent, cover the treatment of severe or fistulised Crohn's disease after failure of steroids or other immunosuppressants. (2) The clinical file includes two comparative placebo-controlled trials with a total of about 200 patients. It is not clear that all the patients included had severe Crohn's disease, and not all had previously been treated with steroids and/or azathioprine. (3) One trial showed that infliximab monotherapy (optimum dose unknown) in patients with mild to moderate exacerbation despite previous treatment yielded remission in one-third of patients within a month (versus 4% on placebo). Retreatment every eight weeks led to sustained remission after ten months in approximately one-quarter of patients. Some patients who did not respond to the first infliximab injection improved after a second one. (4) A trial involving patients with Crohn's disease and enterocutaneous fistulae showed that infliximab healed the fistulae in one third of patients, most of whom had not responded to conventional treatment. (5) Data on the adverse effects of infliximab are limited. A risk of potentially severe infection has been established, especially when infliximab is combined with another immunosuppressive drug. The risks of malignancy, delayed hypersensitivity and autoimmune disorders must be better assessed by close pharmacovigilance. PMID- 11475491 TI - Rofecoxib: new preparation. A disappointing NSAID analgesic. AB - (1) Rofecoxib, a nonsteroidal antiinflammatory drug, is licensed in France for symptom relief in osteoarthritis. It is promoted by MSD-Chibret as a "specific inhibitor of type 2 cyclooxygenases (COX-2)". (2) The clinical dossier includes trials versus other antiinflammatory drugs, but the reports available are generally vague. Rofecoxib has not been compared with paracetamol. (3) In these trials rofecoxib 12.5-25 mg/day was no more effective than the comparators (ibuprofen or diclofenac) used at maximal recommended doses. (4) Relative to other nonsteroidal antiinflammatory drugs prescribed at high doses to selected patients in the controlled conditions of clinical trials, rofecoxib only moderately reduces the risk of severe gastrointestinal reactions (1.3% versus 1.8% after a year of treatment) and dyspeptic disorders (23.5% versus 25.5%). (5) Questions persist on other adverse effects, especially those potentially affecting the kidneys and heart. (6) Rofecoxib is subject to the same precautions (pregnancy, interactions, etc.) as other nonsteroidal antiinflammatory drugs. PMID- 11475492 TI - Treatment of osteoarthritic pain: systemic drug therapy is not always necessary. AB - (1) The treatment of osteoarthritis is symptomatic, and relies first and foremost on physical therapy. (2) Paracetamol is the first choice when systemic drug treatment is deemed necessary. PMID- 11475493 TI - Temozolomide and anaplastic astrocytoma: new indication. No clear proof of efficacy. AB - (1) Temozolomide, a cytotoxic agent, was recently licensed in France for treating patients with anaplastic astrocytoma who are in relapse or progression after standard therapy. (2) The clinical dossier contains only one non comparative trial. (3) In this trial, 111 patients with anaplastic astrocytoma or oligoanaplastic astrocytoma had not all had the standard treatment with surgery, radiotherapy and chemotherapy based on nitrosourea plus procarbazine. In the subgroup of 54 patients who met these criteria, median global survival on temozolomide was 16 months, or 31 months from the start of initial treatment, which was no better than survival before the introduction of temozolomide. (4) The adverse effects of temozolomide include gastrointestinal disorders (nausea, vomiting, constipation), headache, and haematological disorders. PMID- 11475494 TI - Palivizumab in prevention of bronchiolitis: new preparation. Moderate efficacy in some infants. AB - (1) RSV infection, the main cause of bronchiolitis, can necessitate hospitalisation, especially of infants at risk, i.e. those with a history of prematurity or bronchodysplasia. No drug prevention has been available. (2) Palivizumab, a monoclonal antibody directed against respiratory syncytial virus (RSV), is now marketed for preventing respiratory tract infection by RSV in certain infants. (3) The evaluation dossier barely answers the questions raised by the use of this drug. (4) The results of six trials suggest that the optimal dose is 15 mg/kg palivizumab by monthly injection throughout the seasonal epidemic period. (5) A double-blind trial in 1 502 infants either aged less than 6 months and born prematurely (35 weeks of gestation or earlier), or aged under 2 years with a history of bronchopulmonary dysplasia, has shown that, relative to a placebo, palivizumab reduces the hospitalisation rates by 5% in absolute values. It does not influence mortality or the need for mechanical ventilation. (6) Given the lack of relevant trials, we do not know if palivizumab is effective in infants with immunodeficiency or congenital heart diseases. We do not know, either, whether the definition of groups at risk used in the only relevant trial is appropriate. (7) No serious adverse effects attributable to palivizumab were reported in clinical trials. (8) Treatment with palivizumab is costly. PMID- 11475495 TI - RSV infection and bronchiolitis: who qualifies for prevention? AB - (1) Approximately 80% of cases of bronchiolitis appear to be due to respiratory syncytial virus (RSV). (2) Overall, about 1% of cases of bronchiolitis lead to hospitalisation. This rises to about 25% in children with a history of severe prematurity, bronchopulmonary dysplasia or congenital heart disease. PMID- 11475496 TI - Enoxaparin and prevention of venous thrombo-embolism in medical units: new indication. No tangible benefit. AB - (1) In France, enoxaparin 40 mg has been granted a license extension to cover the prevention of deep vein thrombosis in patients confined to bed for acute medical reasons and who have at least one other risk factor. (2) In this indication the clinical file is based on one placebo-controlled trial with methodological weaknesses, such as the use of a surrogate end point (phlebography) and exclusion of a large number of patients from analysis. (3) In this trial fewer than 1% of the 1 102 patients enrolled had a clinical venous thrombosis in the placebo group. Enoxaparin was no more effective than the placebo on clinical outcome measures. (4) Massive bruising at the injection site was more common on enoxaparin than on the placebo. (5) Two trials versus unfractionated heparin, one of which involved nearly 1 000 patients, also failed to show any superiority of enoxaparin over placebo. (6) The cost of enoxaparin in this indication is unjustified, given the lack of proven benefit. PMID- 11475497 TI - Pergolide and Parkinson's disease: new preparation. No clear benefit. AB - (1) Levodopa is the cornerstone of therapy for Parkinson's disease, and bromocriptine is the reference drug for patients who develop motor complications on levodopa. (2) Pergolide, a dopamine agonist, is now marketed in France for the treatment of motor complications associated with levodopa therapy. (3) Four trials comparing pergolide with bromocriptine have been published. The methodological quality of these trials varies, and their published reports often lack detail. (4) Taken together, these trials fail to demonstrate that pergolide provides a tangible clinical advantage over bromocriptine. (5) Pergolide has not been compared with other dopamine agonists in double-blind trials. (6) Pergolide has the same safety profile as other dopamine agonists. PMID- 11475498 TI - Pregnancy and mefloquine prevention of malaria. AB - (1) Mefloquine is teratogenic in animals at high doses. (2) A prospective cohort study of several hundred women exposed to mefloquine at prophylactic doses (250 mg per week) in early pregnancy showed no increased risk of malformations. (3) Exposure to mefloquine in early pregnancy is not an indication for termination. (4) Data on exposure during the second or third trimester are reassuring. According to the World Health Organisation (WHO), the risk-benefit ratio of mefloquine prophylaxis is favourable in pregnant women from the fourth month of pregnancy onwards in areas of group III chemoresistance. (5) In France, mefloquine is contraindicated throughout pregnancy. In other countries mefloquine is approved from the first trimester to cover unavoidable visits to areas of group III chemoresistance. PMID- 11475499 TI - Drug-induced nightmares. AB - (1) A wide variety of drugs have been implicated in nightmares, often on inadequate evidence. (2) Recurrent nightmares can be induced by many drugs, and not only agents with psychotropic or neurological effects. PMID- 11475500 TI - Glaucoma and oral corticosteroids. AB - (1) A case-control survey of patients over 65 taking oral steroids has shown a small rise in the risk of elevated intraocular pressure and glaucoma. (2) It seems advisable to monitor intraocular pressure in such patients, especially if they have a family history or other risk factors. PMID- 11475501 TI - Trial results must be accessible. Clinical research data, a public asset. PMID- 11475502 TI - Think INN, prescribe INN, dispense INN: good professional practice. AB - (1) International nonproprietary names (INN) for drugs were invented about fifty years ago, under the aegis of the World Health Organisation, to provide a common language for health professionals and patients worldwide. (2) No country forbids INN prescriptions. Some countries actively recommend using INNs. (3) INN prescription empowers prescribers and pharmacists in their choice of treatment. (4) The choice between a prescription based on the INN or the brand name will depend on the type of treatment (short term or chronic), the nature of the drug (especially its therapeutic margin) and any specific risks related to the patient (age, disease condition, allergy, and adherence). (5) A pharmacist's decision to dispense a brand name drug from an INN prescription must be based on usual dispensing precautions. (6) Adopting INN prescribing means having to reflect on one's knowledge of drugs, and to challenge the quality of one's initial and continuing education in pharmacology and therapeutics. The INN system is a means of improving prescribing and dispensing practices: it involves paying more attention to the patient, explaining the treatment in greater detail, and respecting his/her choice. PMID- 11475503 TI - Raloxifene and prevention of vertebral fracture (cont'd): mainly when oestrogen is contraindicated. AB - (1) The three-year results of a clinical trial confirm that raloxifene is the second-line treatment for primary and secondary prevention of vertebral fracture in postmenopausal women at risk of osteoporosis. They also confirm that raloxifene increases the risks of thromboembolism and diabetes. PMID- 11475504 TI - Health care law. PMID- 11475505 TI - Watching out for the mentally ill. PMID- 11475506 TI - Sex, lies, and insurance coverage? Insurance carrier coverage defenses for sexually transmitted disease claims. PMID- 11475507 TI - Logical analysis: a method of examination of expert medical opinion through the basic logic of medical reasoning. PMID- 11475508 TI - Evolving limitations on coverage for AIDS: implications for health insurers and employers under the ADA and ERISA. PMID- 11475509 TI - Credentialing liability in the managed care arena. PMID- 11475510 TI - Extending coverage of the Americans with Disabilities Act to individuals with Attention Deficit-Hyperactivity Disorder: a demonstration of inadequate legislative guidance. PMID- 11475511 TI - Differences in access to health care among the moderate-- and low--income population across urban areas. PMID- 11475512 TI - Nearly one-fifth of urban Americans lack health insurance. PMID- 11475513 TI - African Americans remain uninsured at higher rates than whites despite their increasing job-based coverage. PMID- 11475514 TI - Disparity in job-based health coverage places California's Latinos at risk of being uninsured. PMID- 11475515 TI - Declining Medi-Cal coverage leads to increasing uninsured rate among California's Asian Americans and Pacific Islanders. PMID- 11475516 TI - Noncitizen children's rising uninsured rates threaten access to health care. PMID- 11475517 TI - 1.6 million California children are uninsured. PMID- 11475518 TI - Benefits for substance abusers repealed: impact in California. PMID- 11475519 TI - African Americans and Latinos are least likely to receive high-tech heart treatments. PMID- 11475520 TI - Family income and health insurance coverage for women in California. PMID- 11475521 TI - Welfare reform could add 25,000 uninsured L.A. County residents per year. PMID- 11475522 TI - 38% of California's nonelderly Latinos are uninsured. PMID- 11475523 TI - Reforms to cover some, but not all, of California's uninsured children. PMID- 11475524 TI - One in every three young men in California is uninsured. PMID- 11475525 TI - The consequences of being uninsured in California. PMID- 11475526 TI - New estimates find 400,000 children eligible for healthy families program. PMID- 11475527 TI - Barriers and benefits of managed care for low-income women. PMID- 11475528 TI - Women and managed care in California: an examination of selected services. PMID- 11475529 TI - California immigrants have mostly lower rates of disability and use of disability services than state's U.S.-born residents. PMID- 11475530 TI - Focus groups suggest new strategies are needed to reach uninsured children in low income families. PMID- 11475531 TI - Postoperative proprionibacterium acnes endophthalmitis. PMID- 11475532 TI - The genome as a drug target: sequence specific minor groove binding ligands. AB - The ability to target defined sequences on the DNA molecule would be of enormous benefit to the treatment of human disease. Towards this goal much research has been invested in examining the DNA binding and biological mechanisms of action of sequence selective minor groove binding ligands. These compounds act in a variety of ways to inhibit gene expression and DNA replication and also alter nuclear architecture. Concomitant with this, minor groove adducts formed by certain compounds are inefficiently removed by cellular DNA repair systems and are extremely cytotoxic. Additionally compounds targeting A.T rich DNA sequences have found clinical use in the treatment of particular parasitic infections. PMID- 11475533 TI - The DNA methylation machinery as a therapeutic target. AB - Pharmacology and therapeutics have traditionally focused on altering the activity of specific proteins that play an important physiological role either to counteract disease processes or to achieve changes in physiology that are of benefit to the patient. The explosion in our understanding of gene expression programs opens up new horizons for pharmacological intervention. Key processes reversibly controlling genetic programs are attractive drug targets. DNA methylation is a fundamental feature of genomes and the control of their function and is therefore a candidate for pharmacological manipulation that might have important therapeutic advantage. This review argues that DNA methylation plays an important role in the control of genomic processes, suggests how the DNA methylation machinery is involved in cancer, identifies the enzymatic processes that are a target for drug intervention, proposes potential therapeutic utilities for agents that manipulate the DNA methylation machinery and discusses novel drugs that target the DNA methylation machinery. PMID- 11475534 TI - Oligonucleotide-based gene therapy for cardiovascular disease: are oligonucleotide therapeutics novel cardiovascular drugs? AB - Gene therapy is emerging as a potential strategy for the treatment of cardiovascular disease such as restenosis after angioplasty, vascular bypass graft occlusion, transplant coronary vasculopathy, for which no known effective therapy exists. One strategy for combating disease processes has been to target to the transcriptional process. Two approaches have been used to accomplish this. One is the use of antisense that is complimentary to the mRNA of interest. The second approach is the use of ribozymes, a unique class of RNA molecules that not only store information but also process catalytic activity. Ribozymes are known to catalytically cleave specific target RNA leading to degradation, whereas antisense inhibit translation by binding to mRNA sequences on a stoicheometric basis. Theoretically, ribozymes are more effective to inhibit target gene expression. Especially, the application of DNA technology such as antisense strategy to regulate the transcription of disease-related genes in vivo has important therapeutic potential. More recently, transfection of cis-element double stranded (ds) oligodeoxynucleotides (ODN) (= decoy) as a powerful tool in a new class of anti-gene strategies for gene therapy has been reported. Transfection of ds ODN corresponding to cis sequence will result in the attenuation of authentic cis-trans interaction, leading to the removal of trans factors from the endogenous cis-elements with subsequent modulation of gene expression. This "decoy" strategy is not only a novel strategy for gene therapy as an anti-gene strategy, but also a powerful tool for the study of endogenous gene regulation in vivo as well as in vitro. In this review, we have focused on the future potential of oligonucleotide (antisense, decoy & ribozyme)-based gene therapy for the treatment of cardiovascular disease. PMID- 11475535 TI - Ginkgo biloba extract (EGb 761) and CNS functions: basic studies and clinical applications. AB - The effects of EGb 761 on the CNS underlie one of its major therapeutic indications; i.e., individuals suffering from deteriorating cerebral mechanisms related to age-associated impairments of memory, attention and other cognitive functions. EGb 761 is currently used as symptomatic treatment for cerebral insufficiency that occurs during normal ageing or which may be due to degenerative dementia, vascular dementia or mixed forms of both, and for neurosensory disturbances. Depressive symptoms of patients with Alzheimer's disease (AD) and aged non-Alzheimer patients may also respond to treatment with EGb 761 since this extract has an "anti-stress" effect. Basic and clinical studies, conducted both in vitro and in vivo, support these beneficial neuroprotective effects of EGb 761. EGb 761 has several major actions; it enhances cognition, improves blood rheology and tissue metabolism, and opposes the detrimental effects of ischaemia. Several mechanisms of action are useful in explaining how EGb 761 benefits patients with AD and other age-related, neurodegenerative disorders. In animals, EGb 761 possesses antioxidant and free radical-scavenging activities, it reverses age-related losses in brain alpha 1 adrenergic, 5-HT1A and muscarinic receptors, protects against ischaemic neuronal death, preserves the function of the hippocampal mossy fiber system, increases hippocampal high-affinity choline uptake, inhibits the down-regulation of hippocampal glucocorticoid receptors, enhances neuronal plasticity, and counteracts the cognitive deficits that follow stress or traumatic brain injury. Identified chemical constituents of EGb 761 have been associated with certain actions. Both flavonoid and ginkgolide constituents are involved in the free radical-scavenging and antioxidant effects of EGb 761 which decrease tissue levels of reactive oxygen species (ROS) and inhibit membrane lipid peroxidation. Regarding EGb 761-induced regulation of cerebral glucose utilization, bilobalide increases the respiratory control ratio of mitochondria by protecting against uncoupling of oxidative phosphorylation, thereby increasing ATP levels, a result that is supported by the finding that bilobalide increases the expression of the mitochondrial DNA-encoded COX III subunit of cytochrome oxidase. With regard to its "anti-stress" effect, EGb 761 acts via its ginkgolide constituents to decrease the expression of the peripheral benzodiazepine receptor (PBR) of the adrenal cortex. PMID- 11475536 TI - Proteases involved in erythrocyte invasion by the malaria parasite: function and potential as chemotherapeutic targets. AB - Malaria places an increasing burden on global public health resources. In the face of growing resistance of the malaria parasite to available antimalarial drugs, there is a need for new drugs and the identification of new chemotherapeutic targets. The malaria parasite has a complex life cycle which includes a number of obligate intracellular stages. Clinical malaria results from cyclic asexual replication of the blood-stage parasite in circulating erythrocytes of the human host. Erythrocyte entry and host cell rupture require the activity of parasite proteases, and these enzymes are, therefore, attractive targets for rational approaches to new drug development. Malarial proteases play a role in at least two distinct aspects of host cell invasion; modification of parasite proteins involved in host cell recognition and entry; and restructuring of the host cell itself, during and following invasion, and in order to allow parasite release from the host cell. This review details recent advances in the identification of these proteases, describes current understanding of their activation and functional role, and discusses their potential as targets for protease inhibitor-based drugs. PMID- 11475537 TI - P-glycoprotein as a drug target in the treatment of multidrug resistant cancer. AB - Multidrug resistance (MDR) is a major obstacle to successful cancer chemotherapy. One important mechanism of MDR involves the multidrug transporter, P-glycoprotein (Pgp), which confers upon cancer cells the ability to resist lethal doses of certain cytotoxic drugs by pumping the drugs out of the cells and thus reducing their cytotoxicity. Pgp belongs to the ATP-binding cassette (ABC) family of transporter molecules which require hydrolysis of ATP to run the transport mechanism. The substrates of Pgp may be endogenous (steroid hormones, cytokines) or exogenous (cytostatic drugs). A number of studies have demonstrated a negative correlation between Pgp expression levels and chemosensitivity or survival in a range of human malignancies. In principle, Pgp mediated drug resistance can be circumvented by treatment regimens that either exclude Pgp substrate drugs or include Pgp inhibitory agents. Experimental studies have demonstrated that certain structural modifications of anthracyclines confer the ability to escape Pgp transport. The therapeutic benefit of Pgp inhibitors as chemosensitizers is currently being explored in phase III clinical trials, and the first promising results have already been reported. Another therapeutic option for Pgp inhibitors has recently evolved as several Pgp inhibitors, many of which are generally low toxic substances, by themselves constrain proliferation and cause cell death by apoptosis in certain MDR cancer cell lines. The dual effect of Pgp inhibitors, targeting MDR cancer cells selectively, may translate into improved efficacy of cancer chemotherapy and perhaps new and less toxic drug treatment strategies in human MDR cancer. PMID- 11475538 TI - What's a fee for? PMID- 11475539 TI - Unilaterally reinforced facebow for asymmetrical correction. PMID- 11475540 TI - The clinical efficiency of self-ligated brackets. PMID- 11475541 TI - Relating the incisors to points A and B in cephalometric, model, and clinical analysis. PMID- 11475542 TI - New, esthetic organic polymer maxillary retainers. PMID- 11475543 TI - Orthodontic intrusion of periodontally involved teeth. PMID- 11475544 TI - Corticotomy-assisted orthodontics. PMID- 11475545 TI - Impact of a population-based medical curriculum on specialty choice. PMID- 11475546 TI - Gaps in health coverage among working-age Americans and the consequences. AB - This paper examines health coverage and access to care among working-age adults using the Kaiser/Commonwealth 1997 National Survey of Health Insurance. One in three (52 million) working-age adults were either uninsured at the time of the survey or had a recent gap in their health coverage in the past two years. Having even a temporary gap in health coverage made a significant difference in access to care. Compared to the elderly, who are continuously covered by Medicare, working-age adults have greater problems paying their medical bills and gaining access to care and are less satisfied with their health insurance coverage. PMID- 11475547 TI - Barriers to follow-up of hypertensive patients. AB - Lack of follow-up care for hypertension adversely affects health in urban communities. The authors designed this study to (1) evaluate the effectiveness of a specialized intervention program for hypertension follow-up and (2) evaluate the associations with loss to follow-up. They evaluated factors related to loss to follow-up to either a routine care medical clinic or a special primary care intervention program (the Competitive Initiative Program [CIP]). They also conducted interviews to provide in-depth information on the barriers to this program. They found that patients referred through the CIP were significantly more likely to receive follow-up care through a primary care provider. Cost of care, long waiting times, lack of physician continuity, and more pressing priorities explained the lack of follow-up care. Despite a program to provide health care at no cost to patients, lack of insurance and worries about cost are described as barriers to adequate follow-up for hypertension treatment. PMID- 11475548 TI - Poverty, comorbidity, and survival of colorectal cancer patients diagnosed in Connecticut. AB - Studies have reported reduced survival rates for colorectal cancer patients in lower socioeconomic status categories, but this finding could be due (at least in part) to higher comorbidity. This study involved 1,219 patients diagnosed with invasive colorectal cancer in 1992 who were reported to the population-based Connecticut Tumor Registry and followed to their death or through the end of 1997. Risk of death was elevated for patients living in census tracts in the highest quintile for poverty rate, independent of comorbidity (as recorded in a hospital discharge database), age, and stage at diagnosis. Patients living in census tracts with a poverty rate of 20 percent or higher had the highest risk of death. The explanation for these findings requires further study, in order to reduce socioeconomic status disparities in survival rates. PMID- 11475549 TI - Tuberculosis cases reported among migrant farm workers in the United States, 1993 97. AB - Migrant farm workers (MFWs) are considered a high-risk group for tuberculosis. MFW tuberculosis cases reported to the Centers for Disease Control and Prevention represented 1 percent of all reported tuberculosis cases from 1993 to 1997. Most of these cases (70 percent) were reported from Florida, Texas, and California. MFW tuberculosis cases were more likely to be male, foreign-born, or Hispanic and to have a history of alcohol abuse and homelessness than were non-MFWs. Most (79 percent) foreign-born MFWs were from Mexico. HIV status was poorly reported, with results available for only 28 percent of MFW and 33 percent of non-MFW cases. Of the MFWs tested, 28 percent were HIV infected, whereas 34 percent of non-MFWs were HIV infected. Twenty percent of MFWs move or are lost to follow-up before completing therapy; these cases pose a management challenge for the nation's tuberculosis control efforts. PMID- 11475550 TI - Homeless women's gynecological symptoms and use of medical care. AB - Information is lacking on homeless women's gynecological symptoms and use of medical care for symptoms. This paper documents and explains gynecological symptoms and conditions and use of medical care in a probability sample of 974 reproductive-age (15-44) homeless women. Two-thirds of women reported symptoms during the previous year; 71 percent of those received medical care for their gynecological symptoms. Pregnancy, drug dependence, more episodes of homelessness, and general physical health symptoms were positively associated with a number of gynecological symptoms. Gynecological symptoms, younger age, better perceived health, and insurance coverage were positively associated with medical care; women reporting recent drug use and rape received less care. These findings support the importance of medical care and other treatment and support services for homeless women, including expanded care during pregnancy and substance abuse treatment. Health insurance coverage and an interruption in the cycle of homelessness also appear vital to women's health. PMID- 11475551 TI - Hispanic versus white, non-Hispanic physician medical practices in Colorado. AB - The purpose of the study was to extend the scope of earlier research on minority physicians attending to the needs of the poor and their own ethnicity by contrasting practice characteristics of Hispanic doctors in Colorado with those of their white, non-Hispanic counterparts. It was found that Hispanic physicians spent more hours per week in direct patient care, were more likely to have a primary care specialty, and were less often specialty board certified than white, non-Hispanic doctors. Hispanic generalists established practices in areas in which the percentages of the population that were (1) below poverty level, (2) Hispanic, (3) Hispanic and below poverty level, and (4) white, non-Hispanic, and below poverty level were greater than in areas in which white, non-Hispanic primary care physicians practiced. These findings argue for special provision to admit ethnic minorities to undergraduate and graduate medical education programs. PMID- 11475552 TI - Postadmission disparities in nursing home stays of whites and minority elderly. AB - This study examined the type of nursing home stay of white and minority elderly and investigated whether minority differences in nursing home admissions were extended to the type of stay, and if so, what accounted for those differences. African American/white and Latino/white elderly were compared with respect to stays of (1) longer than three months, (2) shorter than three months and discharged alive (recovery), and (3) shorter than three months and deceased (terminal). African American elderly in nursing homes were significantly less likely to have recovery stays than were white elderly. Multivariate analysis revealed that this difference was due to Medicaid coverage. Overall, elderly with Medicaid coverage were more likely to make long-term stays in nursing homes. The findings highlight the importance of Medicaid coverage for short-term recovery nursing home stays. Reductions of benefits or restrictions in eligibility would negatively and disproportionately affect the nursing home stays of elderly African Americans. PMID- 11475553 TI - [Drug resistance in lung cancer]. PMID- 11475554 TI - [Comparison of types and drug sensitivity of bacteria isolated from sputum and bronchoscopic material. Analysis of results from routine diagnostic tests from 1995-1997]. AB - Proper identification of the organism and determination of their sensitivity to antimicrobial agents, is the most important part of the management of patients with symptoms of respiratory system infections. In the case of pulmonary infections, the utility of sputum for diagnosing the cause of lower respiratory tract infections seems to be limited. In the patients receiving antibiotics, there is a high rate of colonization of the upper respiratory tract that may lead to the overgrowth of one or an other pathogen, making it more abundant in the upper than an lower respiratory tract. Also, the patients may be unable to provide adequate lower-respiratory-tract samples. For these reason, more invasive procedures have been recommended. These include bronchoscopy with bronchoalveolar lavage (BAL) sampling. The purpose of our study was to determine the value of sputum and BAL fluid on the basis of profile bacteria and their drug sensitivity isolated from the patients treated in Nat. Res. Inst. Tuber. and Lung Diseases in Warsaw during period of 3 years (1995-1997). On the basis of analysis of 5926 sputum samples and 3534 bronchoscopic materials we have found that the bacterial pathogens species and drug resistant pattern were similar in sputum samples as in materials of bronchoscopic procedures. The same bacterial species and numbers Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were predominant in both type of clinical materials with near the same frequency. PMID- 11475555 TI - [The effect of small cell lung cancer extension on secretion of interleukin-2 (IL 2) and interferon gamma (IFM gamma) in whole blood culture stimulated with mitogens]. AB - The aim of this study was to assess the role of SCLC extension on II-2 and IFN gamma secretion in whole blood cell cultures stimulated with mitogens. Material consisted of 42 SCLC patients, 19 with extensive and 23 with limited disease. In 22 patients partial or complete regression of tumour occurred after treatment. There was a tendency to lower II-2 and IFN gamma secretion before treatment in patients with extensive disease in comparison to those with limited lesions but this was not statistically significant. The ability to secrete cytokines in 22 patients after partial or complete regression of tumour decreased or increased but median values after treatment were not statistically different from those before treatment. PMID- 11475556 TI - [Evaluation of the value of 2 methods for measuring levels of allergen specific IgE antibodies in serum--Phadebas RAST I immuno CAP System]. AB - Two diagnostics sets Pharmacia--Phadebas RAST and Immuno CAP System determining the concentration of specific IgE antibodies in blood serum were compared with each other and with skin prick tests (SPT). Two common alergens--Dermatophagoides pteronyssinus and Dermatophagoides farinae were evaluated. Analysis of ROC curves shows that maximum specificity and sensitivity was obtained when the cut off value of RAST and CAP reached to 4.9 PRU/ml and 7.3 kU/l respectively. Our date shows the good correlation between RAST,CAP and SPT. PMID- 11475557 TI - [Levels of CC-chemokine (MCP-1 alpha, MIP-1 beta) in induced sputum of patients with chronic obstructive pulmonary disease and patients with chronic bronchitis]. AB - BACKGROUND: MCP-1, MIP-1 alpha and MIP-1 beta are CC-chemokines, which act as chemoattractants for inflammatory cells like macrophages, lymphocytes and eosinophils. These cells are known to be important for development of chronic obstructive pulmonary disease (COPD) and chronic bronchitis (CB). METHODS: Sputum was obtained from 14 patients with COPD, 12 patients with CB and 14 healthy persons by means of the sputum induction method. The MCP-1, MIP-1 alpha and MIP-1 beta levels were measured in induced sputum via ELISA method. RESULTS: MIP-1 alpha levels in sputum were significantly higher in CB patients compared to healthy persons (p = 0.01). Median and (range) were following: CB patients--22.7 pg/ml (9.2-95.9 pg/ml), control subjects--17.5 pg/ml (0-27.1 pg/ml). The MIP-1 beta levels in sputum were significantly higher in COPD patients compared to healthy persons (p = = 0.003). Median and (range) were following: COPD patients- 173.2 pg/ml (30.6-1880 pg/ml), control subjects--19.0 pg/ml (0-570.5 pg/ml). No significant differences were detected among studied groups for MCP-1 levels in induced sputum. There was positive correlation in CB group between levels of MIP 1 beta and number of eosinophils in gram of sputum (r = 0.81 and p = 0.001). CONCLUSIONS: MIP-1 alpha maybe important for development of chronic bronchitis and MIP-1 beta for development of chronic obstructive pulmonary disease. MIP-1 beta is probably a chemoattractant for eosinophils in patients with chronic bronchitis. PMID- 11475558 TI - [Levels of CEA, antibodies against mycobacterial antigens and ACE activity in serum and in BALF in patients with sarcoidosis, tuberculosis and lung cancer- preliminary results]. AB - Assessment of CEA concentration, levels of the antibodies against mycobacterial antigens A60 and 38 kDa and ACE activity in serum and in BALF of the patients with sarcoidosis (n = 8), tuberculosis (n = 13) and lung cancer (n = 10) was performed. Nine healthy volunteers were referred to as the control group. Sarcoidosis and lung cancer were confirmed histopathologically. Diagnosis of tuberculosis was stated using the radiometric Bactec 460 system. Serum and BALF CEA concentration and IgG levels against A-60 and 38 kDa were measured by ELISA: IMx, Immunozym and Pathozyme, respectively. ACE activity was assayed by the spectrophotometric method. Statistically significant (p < 0.01) increase in BALF ACE activity in patients with sarcoidosis compared to each group was observed. Serum level of antibodies against 38 kDa and serum ACE activity were specific for tuberculosis and sarcoidosis respectively, but their sensitivity were very poor. Serum CEA concentration was fairly specific for lung cancer but its sensitivity was not satisfactory. CONCLUSIONS: 1. Determination of BALF-ACE activity may be helpful in diagnosis of sarcoidosis but it is recommended to continue the study on more numerous groups of patients taking into consideration polymorphism in the ACE gene. 2. High level of antibodies against 38 kDa and ACE activity in serum are specific for tuberculosis and sarcoidosis respectively but both tests have restricted application because of their poor sensitivity. 3. Serum CEA concentration could be considered only as an auxiliary test in the diagnosis of lung cancer. PMID- 11475559 TI - [Plastic bronchitis and mucoid impaction--uncommon disease syndromes with expectoration mucus plugs]. AB - Mucoid impaction and plastic bronchitis are relatively rare disorders caused by the formation of obstructive airway plugs. We observed from February 1999 to June 2000 seven patients with mucoid impaction and one with plastic bronchitis. In the group of mucoid impaction there were 4 patients with bronchial asthma and 3 without history of lung disease. At the admission to hospital all patients suffered from cough, chest pain and effort dyspnea. Two of them expectorated during cough "bronchial casts". The chest X-ray of 5 patients revealed atelectasis of one of the lung's lobes and diffuse opacities in 2 others. In 4 cases during bronchoscopy one bronchus and in another three--numerous bronchi were obstructed with mucoid casts. Removing of the casts caused both the improvement of the patients' condition and withdrawal of atelectasis in 4 cases. In 5 patients the final diagnosis was allergic bronchopulmonary aspergillosis and in two mucoid impaction in the course of asthma without aspergillosis. Plastic bronchitis was observed in 44 years old man, who expectorated white, branching, bronchial casts for three months. On admission he was in respiratory failure. The chest X-ray revealed diffuse alveolar infiltrates and HRCT glass-ground opacities in both lungs and bronchiectasis in the middle lobe. The bronchofiberoscopy disclosed diffuse tracheobronchitis with casts occluding the middle lobe bronchus. Microscopic examination of the removed casts showed aggregates of mucus, macrophages, neutrophils and cells of respiratory epithelium. Precipitins against Aspergillus fumigatus were not found. Staphyloccocus coagulase (-) was cultured from urine and sputum specimens. We administered Vancomycin with Netylmycin, acetylocysteine, oxygen therapy and humid inhalation and the patient recovered. HRCT made six months after admission revelated total withdrawal of glass-ground opacities. The pathogenesis of plastic bronchitis in this case was unclear. PMID- 11475560 TI - [Extrapulmonary manifestations of sarcoidosis as the cause of primary symptoms--4 case reports]. AB - Sarcoidosis is a multisystem disorder most frequently presenting with hilar lymphadenopathy, pulmonary infiltrations, ocular and skin lesions. However pulmonary manifestations typically dominate, any organ can be affected. Sometimes leading symptoms are caused by extrapulmonary manifestation of the disease, and together with the absence of typical picture in chest radiographs may be confusing for the physicians. We present 4 cases of proven sarcoidosis in which leading symptoms were caused by sarcoidal involvement of different organs (liver, spleen, heart and skin) without typical changes in the lung. In all cases multiorgan involvement was documented and disease was successfully treated. PMID- 11475561 TI - [Atopy with recurrent wheezy bronchitis in children]. AB - Childhood asthma and wheezy bronchitis are the most frequent chronic diseases of childhood. Unfortunately their clinical symptoms are similar--which makes it difficult to distinguish between the two, and therefore decide on proper treatment of patients. The aim of the study was to establish the parameters leading to right diagnosis. The study was performed in 50 children aged 3-7 years with recurrent wheezy bronchitis. All patients underwent allergological examinations (skin tests with inhaled allergens, personal and family history and serum total and specific IgE levels). 42 of them were tested for ventilatory parameters with bronchodilatation test. Three features of atopy were found in 21 (42%) patients, two features in 7 (14%) patients. In 31 (62%) children at least one feature of atopy was shown. 7 (17%) of the examined children had significant bronchodilatation after salbutamol inhalation. Finally in 24 (48%) of children suffering from wheezy bronchitis, bronchial asthma was diagnosed. The diagnosis was confirmed by antiasthma tic treatment with cromones or inhaled corticosteroids. In great majority (88%) of patients bronchial asthma was atopic. In 23% wheezy bronchitis children not diagnosed with bronchial asthma features of atopy were observed. They are of bronchial asthma risk group. PMID- 11475562 TI - [The evaluation of efficacy and tolerability of salbutamol administered by nebulization in patients with stable severe bronchial asthma]. AB - The aim of the study was the assessment of the effect of salbutamol in nebulization on ventilatory parameters and heart action in patients with stable severe well controlled bronchial asthma. The study was performed in 30 asthmatics (19 females and 11 men) with incomplete reversibility of airflow obstruction after salbutamol inhalation administered via MDI device. The mean age was 47 years, the duration of asthma--18 years. Mean FVC value was 2.46 L (66.6%), FEV1- 1.76 L (56.8%), and MEF50--1.74 L/s (40.4% predicted). The study was performed according to the double-blind crossover method with placebo used. On the first day the reversibility test with 400 mg salbutamol was performed. On the two consecutive days salbutamol (Steri-Neb Salamol 2.5 mg) and placebo in nebulization were randomly administered. Ventilatory parameters were measured before and in 20, 40, 60, 120, 180 and 240 minutes after the nebulization. At the same time points the heart action was assessed by physical examination. The changes in FVC, FEV1 and MEF50 were expressed in absolute values and as a relative increase in relation to predictive value. The significant improvement of measured ventilatory parameters was observed as early as 20 minute after the nebulization. This increase in MEF50 lasted 2 hours, in FEV1--3 hours and in FVC 4 hours. The relative increase in MEF50 was significant higher than the remaining parameters. The significant increase in heart rate was noted after salbutamol nebulization. PMID- 11475563 TI - [Recurrent pneumothorax in a child with Langerhans' cell histiocytosis]. AB - We are presenting a 16 months old boy, who has been previously developing normally, physically very active, and who suddenly developed right-sided pneumothorax with infiltrations in both lungs, initially defined as inflammatory. After placing suction drainage of the pleural cavity and antibiotic administration the child's condition improved quickly. The infiltration changes still remained in lungs arousing a suspicion of fibrocystic changes. During an attempt at changing the drainage into a water one, the symptoms of pneumothorax with hypertension increased again. A minithoracotomy with an edge resection of segment 4 and pleurectomy were performed, relating to the histopathological test of the sample, Largenhans' cell histiocytosis was diagnosed. The symptoms of diabetes insipidus appeared. The treatment according to the program DAL-HX83/90 modified by the Polish Group for Leukaemia and Malignant Lymphoma was introduced. During the treatment inducing remission a pneumothorax occurred two more times. PMID- 11475564 TI - [Undiagnosed double aortic arch causing respiratory disturbances after 15-years]. AB - The double aortic arch is a very rare vascular anomaly belonging to a group so called vascular rings. The pressure of abnormal "doubled" aorta on the trachea and esophagus is a cause of sometimes sever symptoms of the respiratory tract such as dyspnea, stridor or cyanosis and recurrent respiratory infections. The authors present a case of a 15 year old boy who had stridor, cough and recurrent respiratory infections. Due to the symptoms the boy was treated for bronchial asthma. Correct diagnosis of double aortic arch was made after the X-ray examination of the chest with barium in the esophagus, followed by bronchoscopy, angiographic and NMR examinations. Surgery treatment was successful. PMID- 11475565 TI - Private choices and public law: Richard A. Posner's contributions to family law and policy. PMID- 11475566 TI - Surrogacy from the perspectives of economic and civil liberties. PMID- 11475567 TI - A civil liberties analysis of surrogacy arrangements. PMID- 11475568 TI - Private religious hospitals: limitations upon autonomous moral choices in reproductive medicine. PMID- 11475569 TI - Application of the corporate negligence doctrine to managed care organizations: sound public policy or judicial overkill? PMID- 11475570 TI - American Manufacturers Mutual Insurance Company v. Sullivan: "Meta-Analysis" as a tool to navigate through the Supreme Court's "state action" maze. PMID- 11475571 TI - Reprocessing and reusing single-use only medical devices: safe medical practice or risky business? PMID- 11475572 TI - Senioritis: why elderly federal inmates are literally dying to get out of prison. PMID- 11475573 TI - "Synthetic sanity": the ethics and legality of using psychotropic medications to render death row inmates competent for execution. PMID- 11475574 TI - Who is responsible for fraternity related injuries on American college campuses? PMID- 11475575 TI - Bringing the "gospel of life" to American jurisprudence: a religious, ethical and philosophical critique of federal funding for embryonic stem cell research. PMID- 11475576 TI - Infant formulae: how well are they tested? PMID- 11475577 TI - The microbiologic and immunologic basis for recurrent otitis media in children. AB - Otitis media is very common in children. A subpopulation of children, representing 5-10% of the general population, are otitis prone and they experience 4 or more episodes of acute otitis media (AOM) in the first year of life. Nasopharyngeal colonization with the three major middle ear pathogens, S. pneumoniae, nontypeable H. influenzae and M. catarrhalis is frequent in otitis prone children and is directly related to the frequency of AOM. Colonization stimulates the production of mucosal as well as serum antibodies to the pathogens. Specific IgA mucosal antibody limits the duration and frequency of colonization. Serum IgG antibody protects children against the development of otitis media but does not affect colonization. Antibody detected in the middle ear often reflects passive transfer from serum rather than local production. Antibody responses to the three pathogens following AOM are generally reduced in the first 2 years of life and rise rapidly thereafter. There are many different strains of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. Among the different strains, there are heterologous surface antigens and some conserved antigens. Conserved antigens induce broadly protective antibodies while strain specific antigens induce limited protection. Although otitis prone children may display strain specific immunity, they often fail to develop a broadly protective antibody response. This subtle immunologic defect makes them susceptible to recurrent and persistent otitis media. CONCLUSIONS: Otitis media is common. Otitis prone children appear to display a subtle immunologic abnormality that predisposes them to recurrent infections. Recent advances in vaccine development may reduce the frequency of otitis media in the general population but the impact on otitis prone children remains unknown. PMID- 11475578 TI - Bladder-sphincter dysfunction in myelomeningocele. AB - Pediatric urodynamics taught us that detrusor-sphincter dyssynergia creates a bladder outlet obstruction in about 50% of any population of children with myelomeningocele. This functional obstruction causes renal damage due to obstructive uropathy, exactly the same way as a congenital anatomical urethral obstruction does. Pediatric urodynamics also taught us that in children with myelomeningocele pelvic floor activity and detrusor activity can be abnormal (hyperactive or inactive) completely independent from each other. These insights have changed the management of myelomeningocele. Children with overactivity of the pelvic floor can be singled out at infant age, and started on clean intermittent catherization, to prevent obstructive uropathy and preserve renal function. Children with detrusor overactivity can be singled out too at very early age, and treated with anticholinergics, to prevent irreversible structural damage to the detrusor and preserve normal bladder capacity and compliance. PMID- 11475579 TI - Phaeochromocytoma associated with a de novo VHL mutation as form fruste of von Hippel-Lindau disease. AB - Phaeochromocytomas usually occur sporadically but may be associated with dominant inherited cancer syndromes such as multiple endocrine neoplasia type 2 (MEN 2), von Hippel-Lindau disease (VHL) and type 1 neurofibromatosis. We report on a boy presenting at age 8 years with an isolated benign phaeochromocytoma of the left adrenal. Three years later a second adrenal phaeochromocytoma was diagnosed on the right side and removed. His family history was negative. Genetic analysis did not show a mutation in the MEN 2 susceptible proto-oncogene rearranged during transfection; however, we found a germline missense mutation in the VHL gene (nucleotide 695 G to A transversion) which has been described only twice before in the literature. Both parents had normal (wild type) VHL copies indicating that our patient had a de novo germline VHL mutation. Careful clinical evaluation of the patient at 18 years did not reveal any other manifestations of VHL disease. CONCLUSION: Carriers of von Hippel-Lindau germline mutations can present with a form fruste of von Hippel-Lindau disease presenting initially with unilateral phaeochromocytoma and therefore mutation analysis should be carried out. PMID- 11475580 TI - Clinical significance of the serum surfactant protein D and KL-6 levels in patients with measles complicated by interstitial pneumonia. AB - To examine the value of surfactant protein D and KL-6 as markers for the diagnosis and the severity of interstitial pneumonia caused by measles infection, surfactant protein D, KL-6 and lactic acid dehydrogenase were measured serially in three patients with measles complicated by interstitial pneumonia as compared to ten measles infected patients without interstitial pneumonia. The serum surfactant protein D and KL-6 levels were higher in patients with measles and interstitial pneumonia as compared to those with measles without interstitial pneumonia. In patients with measles and interstitial pneumonia, the respiratory distress and the alveolar-arterial oxygen differences improved after steroid pulse therapy while the serum surfactant protein D level decreased dramatically under the cut-off level and earlier than the KL-6 level. On the contrary, the serum KL-6 level increased transiently and it took longer to decrease below the cut-off level as compared to the pattern observed for serum surfactant protein D. The serum lactic acid dehydrogenase level changes were between those of the surfactant protein D and KL-6 levels. CONCLUSION: Surfactant protein D and KL-6 are easily measured and useful markers for the diagnosis of interstitial pneumonia caused by measles infection. Early decrease of surfactant protein D contrasts with the transient increase of KL-6 levels after prednisolone pulse therapy. PMID- 11475581 TI - Who are the adolescents who stop smoking? AB - In most industrialised countries, the number of adolescent smokers has increased dramatically during the past several years. Encouraging smokers to quit constitutes one of several methods to control smoking. To improve our understanding of quitting during adolescence and help professionals address the specific attributes and needs of those who quit, we compared the characteristics of youth who report having quit (former smokers, FS) with those of both "regular" smokers (RS) and nonsmokers (NS). As part of the Swiss Multicentre Adolescent Survey on Health, anonymous self-administered questionnaires were distributed to a national representative sample of 9268 15- to 20-year-old teenagers. A multiple logistic regression analysis showed FS to differ from NS for the following variables: suicide attempt (odds ratio, OR 2.10); sleeping difficulties (OR 1.67); delinquency (1.61); assault (OR 1.55); painting of graffiti (OR 1.52); shoplifting (OR 1.51); dissatisfaction with academic choices (OR 1.46); suffering from back problems (OR 1.31); belonging to a sports club (OR 0.78); receiving good marks at school (OR 0.75); and having a positive future expectation with regard to health (OR 0.74). FS differed from RS for the following variables: practising sport (OR 2.18); scouting (OR: 1.97); shoplifting (OR 0.63); tardiness (OR 0.63 and having many friends (OR 0.59). CONCLUSION: Former smokers resemble regular smokers as far as mental health and general health are concerned, whereas they behave similarly to nonsmokers in the field of social and sports activities. PMID- 11475582 TI - Congenital neuroblastoma mimicking early onset sepsis. AB - A newborn girl presented with symptoms of severe early onset sepsis but also with systemic hypertension (SH) at age 3 h. Plasma catecholamine (CAT) levels were extremely elevated, reflecting increased release of CAT from a congenital neuroblastoma (NB). Clinical symptoms at time of admission were: prolonged capillary refill (5 s), tachycardia, tachydyspnoea, metabolic acidosis (pH 7.17, lactate 11.8 mmol/l), fever (38.4 degrees C) and SH: 90/50/65 mmHg (systolic/diastolic/mean). The infant experienced organ failure (lung, heart, liver). A retroperitoneal dumbbell tumour was detected. Plasma CAT levels at age 15 h were: noradrenaline 219 nmol/l; adrenaline 13 nmol/l; and dopamine 65.3 nmol/l. SH responded to intermittent alpha-adrenergic blockage. CAT-related symptoms ceased within 1 week. The intraspinal NB was surgically removed when cord compression became symptomatic. The neurological and developmental state is normal at age 17 months. The abdominal NB regressed spontaneously. CONCLUSION: A neuroblastoma should be considered in newborn infants presenting with a shock like condition together with systemic hypertension. PMID- 11475583 TI - Neurofibromatosis type 2 diagnosed in the absence of vestibular schwannomas. A case report and guidelines for a screening protocol for children at risk. AB - A 5-year-old girl presented with multiple tumours of the central nervous system. As on the first MRI scan bilateral vestibular schwannomas were not detected due to their small size, she initially did not meet the criteria for neurofibromatosis type 2 (NF2), although her clinical symptoms were highly suggestive for the diagnosis. Using molecular studies, a mutation in the NF2 gene was found confirming the clinical suspicion at an early age and indicating the value of molecular analysis. Follow-up MRI 3 years later demonstrated bilateral vestibular schwannomas more clearly, since they had increased in size. CONCLUSION: In children, magnetic resonance imaging can be inconclusive for the diagnosis of neurofibromatosis type 2, since very small vestibular schwannomas may be missed. In these cases molecular studies may provide additional evidence for the diagnosis. We propose guidelines for a screening protocol for children at risk for having neurofibromatosis type 2. PMID- 11475584 TI - Carbamazepine in phenobarbital-nonresponders: experience with ten preterm infants. AB - Carbamazepine is a standard anticonvulsant in children and adults. Until now there is only little information available on its use in neonates. We investigated the oral administration of carbamazepine in refractory neonatal seizures treated with phenobarbital. Ten preterm infants (gestational age 23 + 6 34 + 6 weeks, birth weight 640 g-3080 g) with neonatal seizures were refractory to a primary therapy with phenobarbital. All patients subsequently received carbamazepine exclusively as a second choice anticonvulsant. A daily dose of 7-23 mg/kg carbamazepine was administered orally in two to three aliquots. All patients reached therapeutic plasma drug levels (3-12 mg/l; 13-50 mumol/l). In nine out of ten patients (complete group of small preterms with gestational age under 30 weeks and weight less than 1000 g), therapeutic success was excellent. Carbamazepine was continued for 1-5 months. After termination of therapy no further seizures occurred, also on EEG recordings. Finally, no carbamazepine induced adverse effects were observed. CONCLUSION: This is the first report on the use of carbamazepine in small preterm infants. Carbamazepine may provide a useful and effective oral maintenance therapy in the management of neonatal seizures in these patients. PMID- 11475585 TI - Pacifiers during sleep and sudden infant death. PMID- 11475586 TI - Endogenous endophthalmitis and necrotising pneumonia caused by Klebsiella pneumoniae in a child with beta-thalassaemia major. AB - A case beta-thalassaemia major complicated with Klebsiella pneumoniae endogenous endophthalmitis is presented to increase physician awareness of the association of the two clinical conditions. Severe morbidity including subretinal abscess and retinal detachment may develop despite aggressive intravenous and intravitreal ceftriaxone therapy, along with vitrectomy and external drainage. PMID- 11475587 TI - An ultra premature baby of 290 g birth weight needed more than 500 mg/kg of calcium and phosphorus daily. AB - Ultrapremature babies accompanied by intrauterine growth retardation may require supplemental calcium and phosphorus far above the recommended doses. PMID- 11475588 TI - Alpha-sarcoglycanopathy previously misdiagnosed as Duchenne muscular dystrophy: implications for current diagnostics and patient care. AB - Differential diagnosis of limb-girdle muscular dystrophy, including alpha sarcoglycanopathy and Duchenne muscular dystrophy, is impossible to acheive on clinical grounds alone; therefore immunohistology, Western blotting and molecular genetic analysis are manadatory for a correct diagnosis. The patient's genotype with a hitherto unknown mutation (Tyr134STOP) in exon 5 adds to the growing spectrum of mutations in the alpha-sarcoglycan gene. PMID- 11475589 TI - Elevated KL-6 levels in fatal measles pneumonia. AB - We measured KL-6 concentrations in serum samples from measles patients with various forms of pneumonia and abnormally elevated levels of KL-6 were found exclusively in four fatal cases of pulmonary insufficiency. PMID- 11475590 TI - Acute idiopathic scrotal oedema: four cases and a short review. AB - We report four cases of acute idiopathic scrotal oedema, an underreported cause of acute painless or moderately painfull swelling and erythema of the scrotum in young boys. It is a self-limiting disease and to prevent unnecessary surgical exploration or antibiotic therapy, it is very important to distinguish acute idiopathic scrotal oedema from more serious diseases. PMID- 11475592 TI - Health tips. Volunteering and your health. PMID- 11475591 TI - Mild cognitive impairment. Is it the first sign of Alzheimer's disease? PMID- 11475593 TI - New test for intermittent claudication. PMID- 11475594 TI - Moderate activity on a daily basis is healthy. PMID- 11475595 TI - The ABCs of CBC. A common blood test. PMID- 11475596 TI - Endometrial biopsy. Finding the cause of uterine bleeding. PMID- 11475597 TI - Baker's cysts. Common cause of a bulge behind the knee. PMID- 11475598 TI - I'm taking over-the-counter ibuprofen every day for neck pain. Recently, my doctor gave me a prescription for naproxen for my arthritis. Should I be concerned about taking these two together? PMID- 11475599 TI - I've heard germs can be spread by cutting boards. Is this true? PMID- 11475600 TI - [Arthrosonography: from an alternative method to standardized and integrated procedure in diagnostic imaging in rheumatology]. PMID- 11475601 TI - [Technique and value of arthrosonography in rheumatologic diagnosis. 1: Ultrasound diagnosis of the knee joint]. AB - Within the last few years, ultrasonography (US) of joints has attained a firm position for the diagnosis of joint diseases. Degenerative as well as inflammatory changes can be recognized using this method. With new, higher resolution techniques even bone surfaces and tendon textures can be visualized in greater detail. The advantages of sonography are general availability and low costs. When used properly, as a non-invasive procedure US has no detrimental effects on patients. The disadvantages of this technique result from its physical limitations, such as high reflection of US on bone and the negative correlation between resolution and penetration which makes US imaging difficult in deeper regions. The current technical development of probes and imaging processing, however, promises better deep structure imaging in the future. The quality of ultrasound examination always depends on the technical equipment, as well as on the patients' individual tissue constitution and the experience of the physician. It is possible to avoid misinterpretation and to increase the diagnostic value of US by using a standardized technique and professional knowledge of the specific aspects of the method. The significance of qualified education and sufficient training of sonographers is to be stressed. The aim of this article is to deliver a basic contribution to the standardization and quality assurance of joint US and to indicate the value of this method. In addition to the overview the authors propose guidelines for performance and interpretation of joint US. Due to the major significance of the knee joint in rheumatology it was decided to begin the work in this area. PMID- 11475602 TI - Power Doppler sonography in the evaluation and follow-up of knee involvement in patients with juvenile idiopathic arthritis. AB - INTRODUCTION: This study was undertaken to evaluate the role of ultrasound (US), conventional color (CD) and power Doppler (PD) in the detection and quantification of inflammatory signs of the knee in children with juvenile idiopathic arthritis (JIA) and to correlate these findings with patient history, clinical, laboratory and radiological findings. PATIENTS AND METHODS: Thirty patients with JIA who had clinical signs of knee involvement as well as 15 healthy children as a control group where subjected to full clinical examination and laboratory investigations on the same day of US examination. The knee joints were evaluated with plain radiography, US, and color Doppler in 13 patients, while the remaining 17 were assessed with power Doppler. Fourteen patients were subjected to follow-up assessment. RESULTS: A highly significant difference in synovial thickening and cartilage thickness detected by US between JIA affected knees and those of controls (p < 0.0001). Knee effusion was demonstrated in 93% of patients. Synovial vessels were detected by Doppler in 76.7% of patients. A significant correlation was detected between the degree of vascularity detected by PD and knee score (p < 0.05), and JAFAR score (P < 0.05). On comparing the findings of the follow-up with those of the initial examination, a significant positive correlation was detected between the differences in the knee score and those in synovial thickness (p < 0.05), and with the vascularity scale detected by PD (p < 0.05). CONCLUSION: This study suggests the Doppler sonography as a non invasive, low-cost, and readily available tool for the evaluation and follow-up of articular involvement in knees of JIA patients. PMID- 11475603 TI - A method to score radiographic change in psoriatic arthritis. AB - INTRODUCTION: Radiographic features of psoriatic arthritis (PsA) are very characteristic and differ from those observed in rheumatoid arthritis, especially in two aspects: 1) the distribution of affected joints (i.e. DIP joints), 2) the presence of destructive changes and bone proliferation at the same time. A scoring method for PsA, therefore, has to account for these characteristics of PsA. OBJECTIVE: To develop, describe and validate a method for scoring radiographic changes in patients with PsA. DESCRIPTION OF THE METHOD: Forty joints of the hands and feet are scored for destruction and proliferation. In the destruction score (DS) grading on a 0-5 scale is based on the amount of joint surface destruction: 0 = normal, 1 = one or more erosions with an interruption of the cortical plate of > 1 mm with destruction of the total joint surface up to 10%, 2 = 11-25%, 3 = 26-50%, 4 = 51-75%, 5 = > 75% joint surface destruction. The proliferation score (PS) sums up any kind of bony proliferation typical for PsA; graded 0-4: 0 = normal, 1 = bony proliferation of 1-2 mm or bone growth < 25% of the original size (diameter), 2 = bony proliferation 2-3 mm or bone growth 25 50%, 3 = bony proliferation > 3 mm or bone growth > 50%, 4 = bony ankylosis. The DS (0-200) and the PS (0-160) can be summed up to the total score (0-360). VALIDATION OF THE METHOD: To validate the method x-rays of 20 patients with active PsA taken 3 years apart were read twice in pairs, knowing the chronological order but not knowing demographic, clinical or laboratory data of the patients. The data were analyzed with a hierarchical analysis of variance model. RESULTS: There was good agreement between the first and the second reading of the same rater and between the two raters regarding the destruction score. The agreement regarding the proliferation score was lower but still acceptable. The reliability of the method to describe change over time--relation of progression (intra-patient variance) to the measurement error (inter-rater variance)--was 3.9 for the DS, 2.8 for the PS and 4.1 for the total score. The minimal detectable change when the readings of two raters were compared (inter-rater MDC) was 5.8, 5.0 and 4.6%, respectively of the maximum possible score for the destruction, the proliferation and the total score. These data compare very well with the results of standard scoring methods in rheumatoid arthritis. CONCLUSION: We propose a method for scoring radiographic change in psoriatic arthritis which reliably quantifies the progression of the disease seen on radiographs. PMID- 11475604 TI - [Molecular mechanisms of action of gold in treatment of rheumatoid arthritis--an update]. AB - Gold was first used 90 years ago by Robert Koch for the treatment of tuberculosis based on the assumption that rheumatoid arthritis was caused by microbacteria. It soon became clear that this would not explain the action of gold in rheumatoid arthritis, and since then scientists have been struggling to elucidate the mechanisms of gold's action in the treatment of rheumatic diseases. In nearly every area of immunology inhibiting actions of gold could be documented; however, it is still unclear if there is a common denominator or if there are parallel modes of actions which are independent of each other. In any case, also based on recent studies the reactivity of gold compounds with thiol groups appears to the predominant factor. Analyzing the actions of gold in the different phases of an immune reaction suggested that gold plays an important role already in the initiation, namely the uptake and presentation of foreign antigens. Thus, gold is taken up by the macrophages and stored in the lysosomes which are called aureosomes where gold inhibits antigen processing. Especially peptide antigens, which contain sulfur such as cysteine and methionine, are important. Moreover, it could be shown that gold suppresses NF-kappa B binding activity as well as the activation of the I-kappa B-kinase. This mechanism results in a subsequently reduced production of pro-inflammatory cytokines, most notably TNF-alpha, interleukin-1 and interleukin-6. On the subsequent T-cell level, gold has been shown to induce an upregulation of IL-4 mRNA, resulting in a shift of the T-cell population to the Th2 phenotype. Moreover, the activation of T-cells is inhibited. On the effector level, gold inhibits proteolytic enzymes and can result in the destruction of synovial fibroblasts. In conclusion, gold remains one of the most fascinating antirheumatic drugs with multiple modes of actions. The future analysis of molecular mechanisms, especially with regard to signal transduction, will lead to new fundamental knowledge of gold action, possible allowing a further understanding of the pathogenesis of rheumatoid arthritis. PMID- 11475605 TI - [Introduction of the Freiburg Competence Center]. PMID- 11475606 TI - ["Scientific Continuing Education, Sponsoring and Research Cooperation in the Tension Field of the Anti-Corruption Law". Protocol of workshop in Berlin 19 January 2001, 10 a.m. to 3 p.m., Kaiserin Friedrich Foundation]. PMID- 11475607 TI - [Stereoscopic simulation of ear surgery intervention with a novel 3D computer models]. AB - INTRODUCTION: The presentation of the surgical anatomy of the temporal bone by standard anatomical figures is not suitable for otosurgical training. For the comprehension of its complex morphology temporal bone drilling is inalienable. Aim of the present cooperation was to gain an interactive real-3D program for the simulation of specific laterobasal surgical approaches. METHODS: The program was derived from a standard horizontal section of a human temporal bone using a Siemens Somatom Plus 4 Tomograph. The slice thickness was 1 mm, the image matrix was 512. The Voxel-Man-system was used to built up the application for unix workstations. RESULTS: Each step of a surgical approach to the temporal bone can be performed by the present computer model. Calculation in a stereo mode even allows spatial 3D-perception when using red/green glasses. CONCLUSIONS: This program is a novel tool to simulate critical aspects of otosurgical procedures on a computer. Up to now the lack of tactile and kinesthetic information does not allow to renounce individual temporal bone drilling. PMID- 11475608 TI - [Vertigo in suction of open mastoid cavities ("radical surgery cavities")]. AB - BACKGROUND: Vertigo during suction-cleaning of open mastoid cavities is caused by cold stimulation of the vestibular organ. In clinical and model-experimental measurements we tried to estimate the physical background and the dimension of this temperature drop. METHODS: Using different techniques like thermo-probes, thermovision, or additional model experiments temperature changes during suction were measured in 5 volunteers with open mastoid cavities. These registrations were carried out with special regard to the moisture of the cavity-walls. RESULTS: Distinct temperature changes during suction were observed. In moist cavities superficial temperature decrease was much higher than in dry ones. Using additional model experiments, temperature conduction towards e.g. the horizontal semicircular canal could be simulated. DISCUSSION AND CONCLUSIONS: Among other mechanisms like replacement of sucked-off air by colder air, or taking away warm air close to the cavity walls, effects of evaporative cold seem to be very significant. This could be demonstrated by moistening cavity walls in patients as well as in model experiments. The conclusion to keep the cavity walls as dry as possible is already a clinical demand. In this paper, the physical background and the dimensions of thermal effects during suction-cleaning could be elucidated. PMID- 11475609 TI - [Clinical relevance of developmental changes of the endolymphatic sac]. AB - BACKGROUND: Outcome of endolymphatic sac shunt surgery crucially depends on endolymphatic sac morphology. The intraoperative finding of a fibrotic sac, e.g., will lead to therapy-failure. Aim of the present study was to determine possible developmental changes in endolymphatic sac morphology which could imply, that a shunt operation should not be undertaken in older age. METHOD: 46 histological temporal bone serial sections were examined. All sections were taken from the "Wittmaack Temporal Bone Collection". All specimens were obtained from patients with morphologically healthy ears, especially without a history of Meniere's disease or other kinds of vertigo. RESULTS: During the fetal period the endolymphatic sac is a wide cavity that is surrounded by immature embryonal connective tissue. In older age this cavity gradually shrinks, leading to stenosis in about 80% of the cases. The endolymphatic duct, however, remains open. CONCLUSION: The increasing fibrosis of the endolymphatic sac is part of the natural involution of the inner ear. It constitutes a natural limitation for the outcome of endolymphatic sac shunt surgery in the elderly. PMID- 11475610 TI - [Cutaneous malignant melanoma in the area of the head and neck with intermediate tumor thickness: does primary site have prognostic relevance?]. AB - BACKGROUND: Tumor thickness and infiltration of malignant melanoma are the main prognostic factors for recurrence and survival. The sentinel lymph node biopsy may provide a step toward a more individual staging and therapy. It was the aim of this study to investigate the prognostic influence of the primary localization of head and neck melanoma subdivided into scalp, ear, neck, and face. To form a basis for routine sentinel lymph node biopsy in case of intermediate tumor thickness metastatic pattern of the different primary sites were analysed. METHOD: Survival rates depending on primary tumor site of 51 patients with cutaneous malignant melanoma of the head and neck were analysed. Metastatic pattern were evaluated with the help of clinical, intraoperative, pathohistologic data. RESULTS: Scalp and ear melanoma were found to be higher risk lesions with a 5-year survival rate of 28% and 59% respectively. Melanoma of the neck and face had a 5-year survival rate of 75% and 78% respectively. In two cases of ear melanoma lymph node metastases could be demonstrated in unusual sites by skipping the first draining basin. In three cases bilateral metastases could be shown histologically as opposed to clinical prediction. CONCLUSIONS: Localization of cutaneous malignant melanoma of the head and neck is a prognostic factor for survival. Because of the discordancy between clinical prediction and lymphatic drainage pattern sentinel lymph node biopsy improves accuracy of diagnostic and therapeutic procedures. PMID- 11475611 TI - [Comparison between the Eppendorf histograph and the Licox system for determination oxygen partial pressure in tumor tissue]. AB - BACKGROUND: The oxygenation of a tumor is certainly one of the major facts influencing the response to treatment. pO2 measurement in tumor tissue can be done by using the Eppendorf-histograph. In recent years the Licox-system (GMS) was developed for oxygen tension measurement in tissues and fluids under clinical routine conditions and for research work. Aim of the present study was to assess the suitability of the Licox-system in drawing up histograms of pO2 values in tumor tissues. METHOD: Histograms of the distribution of oxygen tension in twelve tumors were prepared with both measuring systems in parallel. Investigations were performed on rats with isotransplanted R1H rhabdomyosarcomas in continuous intravenous anesthesia with Fentanyl and Dormicum. Ventilation with a mixture of air and oxygen was applied through a tracheostoma. Arterial pressure and body temperature were analysed and balanced continuously. RESULTS: No correspondence between data of both measuring methods could be found. CONCLUSIONS: Because of constructional advantages the Eppendorf-histograph seems to be comparatively more suitable in drawing up histograms of oxygen tension tumor tissue. PMID- 11475612 TI - [Tracheostomy valve with integrated cough flap for improving hands-free speech in laryngectomized patients--development and clinical applications]. AB - BACKGROUND: Following successful voice restoration after laryngectomy either by a voice prosthesis, a surgical shunt or microvascular laryngoplasty, a further goal in rehabilitation is the insertion of a tracheostoma valve, which enables the patient to speak without using his fingers for closure of the tracheostoma. One important disadvantage of the tracheostoma valves, which are available today, is the necessity of removal of the valve in case of coughing, because the valve could be thrown from the stoma by the strong air flow during coughing. As many laryngectomies suffer from chronic bronchitis, this coughing problem is one of the reasons why only few patients could be provided with this useful aid. METHOD: At the department of biomedical engineering of the faculty of medicine at the university of Groningen, the Netherlands, 1994 two prototypes of a tracheostoma valve with an integrated cough lid were developed. These devices contain two separate valve systems: the normal speaking valve and a special coughing valve, which opens at a certain air flow and closes automatically after the coughing attack. Thus no manipulations are necessary during coughing, the patient can speak undisturbed. The ADEVA company (Lubeck, Germany) undertook the industrial production of this new type of tracheostoma valve creating different modifications of the prototype #2. PATIENTS: In four series with 6-8 patients per group the modified tracheostoma valves were tested clinically and the occurring faults or lack of correct function eliminated by small changes in the production. RESULTS: Meanwhile a suitable model for routine use is available, which was tested in 30 patients so far. This suitability was achieved by improvements in the valve mechanism, the valve seal and the adjustment mechanisms for the individual pressure level of the speaking and the coughing valve. CONCLUSION: The newly developed tracheostoma valve with integrated coughing lid (Window, ADEVA medical Company, Lubeck, Germany) provides further improvement in speech rehabilitation of laryngectomies. The low acceptance of tracheostoma valves, which enable the patient to speak without using his fingers for closure of the tracheostoma, possibly may be raised by this new aid. PMID- 11475613 TI - [Topographic diagnosis in the area of the head-neck: initial experiences comparing ultrasound panorama images with CT and MRI]. AB - PURPOSE: Due to the restricted width of the ultrasound array, documentation of topographical correlations in B-scan-echography is still difficult. By means of Color SieScape it is possible to produce successive images of the regions of interest by lateral movement of the ultrasound array. Aim of the present study was to evaluate this new method in comparison to CT and MRI. METHOD: 18 patients with tumors in different areas of the head and neck region were examined by CT or MRI and by sonography. Color SieScape was used for documentation. A Sonoline Elegra ultrasound platform with 7.5 MHz linear array was used for the investigations. RESULTS: Because of claustrophoby no MRI-investigation could be done in 4 of the patients. Sonography and Color SieScape documentation was possible in all cases. In addition to the panorama-information of SieScape technique in every investigation by Color SieScape large and very small vessels could be documented in colour. CONCLUSION: By the use of Color SieScape the identification of the topographical correlations is improved. The spatial resolution is comparable to MRI and better than in CT. Investigation of patients who suffer from claustrophobia is much easier by Color SieScape. The imaging of parapharyngical structures could not be improved by Color SieScape. Nevertheless it is the only technique for the coloured documentation of even small vessels and topographic relations. Color SieScape investigations are faster and less expensive than CT or MRI. PMID- 11475614 TI - [Endoscopic multi-color deglutition study]. AB - BACKGROUND: The complexity of the diagnostic/therapeutic problem in ENT-specific as well as in neurogenic dysphagia requires that test foods be offered in a variety of consistencies (viscosities), at different temperatures, and with various tastes. The swallowing assessment is undertaken with different head and trunk postures, using different swallowing techniques. METHODS: A series of test foods of varied viscosities is prepared with different food colorings. It is given to the patient according to an individually-tailored viscosity plan, but usually in the same (repeatable) color sequence of blue-yellow or blue-yellow black, an order which offers optimal color contrasts. RESULTS, CONCLUSIONS: During repeated swallowing assessments, the "multicolor technique" is more useful for endoscopic observation of the pharyngolaryngeal route of the food bolus than the "methylene blue swallow". Food coloring is cheap and it changes neither the taste nor the pH of the food. PMID- 11475615 TI - [Interesting case no. 46. Bronchial foreign body]. PMID- 11475616 TI - [Significance of cytokines and chemokines in infections of the upper respiratory tract]. PMID- 11475617 TI - [Traumatology of the soft tissues of the neck. II]. PMID- 11475618 TI - [Inline skating in school--perspectives from the sports medicine viewpoint]. AB - INTRODUCTION: Children are motivated by new activities in physical education. Inline Skating (ILS) is one of the trend activities of the last years. For the establishment of ILS in school injury risks and the possible preventive transfer in leisure time activities are of interest. The aim of this study was to compare the incidence of injuries in an ILS-program in school with traditional activities of exercise lessons. Furthermore inline skate specific injury incidence in leisure time was evaluated. METHOD: A sample of 47 schools was evaluated by a standardized questionnaire asking for: injury incidence in traditional sports and ILS and the incidence of ILS-injuries in leisure time. RESULTS: The relative injury risk of ILS (% per 100 teaching units) is 14.3%. Basketball e.g. has an injury risk of 16.1%, handball 14.6%, gymnastics 13.9% and other games 9.5%. After the end of the ILS-program 37.3% of the students reported the incidence of an injury due to ILS in leisure time. CONCLUSION: Compared to other activities in physical education ILS shows a moderate risk of injury and may improve the acceptance of active and passive prophylaxis. PMID- 11475619 TI - [Effect of external stabilization aids of the upper ankle joint on sports specific performance with reference to isokinetic strength measurements]. AB - Two questions were investigated: Do ankle devices have any influence on sports performance? Are there any differences between the devices? 31 participants could be included. The average age was 24.5 (+/- 4.1). The participants exercised 5.8 hours/week. We tested 41 ankles without any previous injury. With the Cybex 6000 four parameters were evaluated. Torque maximum for the plantarflexion, range of motion, work in Joule. The ankles were tested with three different ortheses, ankle taping and without any device. The Friedman Test was used to evaluate differences between the five test conditions. All measurements showed significant worse results for the devices and taping. The protective effect of ankle devices is well known. In this investigation we could show a restriction of the performance. Thus it should be decided individually, if an ankle device is useful. PMID- 11475620 TI - [Electronic plantar pressure measurements in different types of moutaineering boots]. AB - Specific footwear has been developed for different, partly newly developed mountain sports. This footwear has very different basic characteristics. This experimental study is dealing with the dynamic parameters of electronic plantar pressure measurements in normal walking using different mountain sport shoes designed for a special purpose in mountaineering. These mountain shoes included a hiking shoe, a trekking shoe and a high-alpine shoe which can take crampons. The electronic plantar pressure measurements were done with the Parotec system of the company Paromed with insoles with hydrocell-sensors. The data obtained during the study were proceeded through a variant analysis. There were no significant differences for impulse and contact time, nevertheless concerning maximal pressure there were significant differences between walking barefoot and walking with the hiking and high-alpine shoe. Even when walking in different mountain shoes the feet conserve their own characteristics and their walking style. The different mountain shoes did not alter most of the measured plantar parameters significantly comparing barefoot walking to walking with the different mountain shoes or comparing walking in the different mountain shoes. PMID- 11475621 TI - [Use of extracorporeal shockwave therapy (ESWT)in sports orthopedics]. AB - From 11/97 to 10/99 409 patients have been treated with the extracorporal shockwave therapy (ESWT) in the sport orthopaedic ambulance. An ultrasound controlled treatment was applied in the low and middle energy range, using energy densities of 0.09 to 0.36 mJ/mm2. The ESWT came into application as an operation avoiding procedure after exhausting the conventional conservative therapy measures. Among the 409 patients, 65 persons were competitive sportsmen. Besides the established indications for the application of the shock wave therapy, this method also has been applied with specific sport orthopaedic indications like achillodynia, patella-top-syndrome, medial shin-syndrome, morbus Schlatter or tendinosis of the shoulder. The results were collected three months after the application of the ESWT using a standardized questionnaire. The performed examination exhibits that for conservatively treated tendinoses of the competitive sportsmen, similarly good therapy results concerning the application of ESWT can be reached compared with the classical orthopaedic shockwave therapy. Therefore, for the treatment of a tendinosis, a shock wave therapy should always be taken into account to avoid long exercise and competition breaks due to operative interventions. PMID- 11475622 TI - [Injury patterns and prevention in World Cup windsurfing]. AB - The aim of the present study was to analyze the injury patterns in professional world cup windsurfing. The one handed jumps and loops are combined with a high risk of injury. 49 world cup professional windsurfers (39 men, 10 women) could be asked regarding their injuries in their professional career. Questionnaires were used in German, English or French. Furthermore the windsurfers were asked about prophylaxis against injuries. 260 injuries could be stated: 43 injuries of the head (17%), 40 injuries of the trunk (15%), 23 injuries of the upper extremity (9%). The majority of the injuries were stated at the lower extremity (n = 154, 59%). As far as the lower extremity is concerned, the strain of the ankle joint was dominating (n = 57). With 22% of all mentioned injuries this was the most common injury and 61% of the windsurfers claimed about an injury of the ankle joint. An injury of the knee was stated in 30 windsurfers, whereby 80% of these injuries were severe injuries of the ligaments or the meniscus. Only 10% of the professionals used a helmet to prevent injuries of the head. The cause of the head injuries are the spectacular jumps like loopings or table tops. The cumulation of injuries of the lower extremity is due to the fixation of the feet in the footstraps. The fixation on the feet leads to extreme rotatory forces in the knee joint in case of a fall. Furthermore the fixation of the feet leads to a high rate of ankle joint strain. In order to prevent injuries a helmet should be used and special footstraps should be developed, which ensure a release of the foot in the danger area. PMID- 11475623 TI - [Application of modern methods of movement analysis for functional diagnosis and risk screening of the athlete]. PMID- 11475624 TI - [Continuing medical education (CME): an old theme that's once again important?]. PMID- 11475625 TI - [Patient safety and errors in medicine: development, prevention and analyses of incidents]. AB - "Patient safety" and "errors in medicine" are issues gaining more and more prominence in the eyes of the public. According to newer studies, errors in medicine are among the ten major causes of death in association with the whole area of health care. A new era has begun incorporating attention to a "systems" approach to deal with errors and their causes in the health system. In other high risk domains with a high demand for safety (such as the nuclear power industry and aviation) many strategies to enhance safety have been established. It is time to study these strategies, to adapt them if necessary and apply them to the field of medicine. These strategies include: to teach people how errors evolve in complex working domains and how types of errors are classified; the introduction of critical incident reporting systems that are free of negative consequences for the reporters; the promotion of continuous medical education; and the development of generic problem-solving skills incorporating the extensive use of realistic simulators wherever possible. Interestingly, the field of anesthesiology--within which realistic simulators were developed--is referred to as a model for the new patient safety movement. Despite this proud track record in recent times though, there is still much to be done even in the field of anesthesiology. Overall though, the most important strategy towards a long-term improvement in patient safety will be a change of "culture" throughout the entire health care system. The "culture of blame" focused on individuals should be replaced by a "safety culture", that sees errors and critical incidents as a problem of the whole organization. The acceptance of human fallability and an open-minded non-punitive analysis of errors in the sense of a "preventive and proactive safety culture" should lead to solutions at the systemic level. This change in culture can only be achieved with a strong commitment from the highest levels of an organization. Patient safety must have the highest priority in the goals of the institution: "Primum nihil nocere"--"First, do not harm". PMID- 11475626 TI - [Validation of an observer-based rating set compared to a standardized written psychological test for the diagnosis of depression and anxiety in a university preadmission test center]. AB - INTRODUCTION: Depression and anxiety can be a major factor of perioperative stress and might contribute to patients dissatisfaction with medical care if they remain unrecognized. There are several methods to diagnose depression and anxiety like standardized written psychological tests or self report scales. Because these tests are not always suitable for routine use in a busy preadmission test center we evaluated an observer-based rating set for the diagnosis of depression and anxiety. MATERIAL AND METHODS: 70 patients of a university hospital preadmission test center were tested with the HADS-D-Test and the observer-based rating set after approval of the institutional review board and written informed consent. Test-data were compared using a logistic regression model and demographic variables were analyzed using t-Test. ANOVA and Pearson correlation. RESULTS: The prevalence of depression in our study population was 11.11% (14.75% in male, 9.76% in female) and the prevalence of anxiety was 7.14% (6.9% in male and 7.32% in female). The correlation between the observer-based rating items and the HADS-D-diagnosis was statistically highly significant. The observer based items "unsteady eye movements" and "general worrisome mood" proved to be especially sensitive for anxiety and the items "sorrowful mood" and "impression of resignation" were sensitive for depression without any influence of the experience of the anesthesiologist. A higher prevalence of depression and anxiety was found in patients with ASA-class III compared to those with ASA-classes I and II while age and type of surgery had no significant influence. CONCLUSIONS: Based on our observations depression and anxiety are a relevant factor of preoperative morbidity assessment. Observer-based items are a reliable tool to detect those patients which might need special assistance and therapy in the perioperative period to reduce stress associated with high preexisting levels of depression and anxiety. PMID- 11475627 TI - [Gastrointestinal atony in intensive care patients. Mechanisms, causes, therapy]. PMID- 11475628 TI - The future of inhalation anaesthesia. Die Zukunft der Inhalationsanasthesie. PMID- 11475629 TI - [Benefit-risk profile of new inhalation anesthetics]. PMID- 11475630 TI - [Cerebral effects of volatile anesthetics--what do we really know?]. PMID- 11475631 TI - [Neuromonitoring--a must for the future?]. PMID- 11475632 TI - [Anesthetic concepts with volatile anesthetics]. PMID- 11475633 TI - [Inhalation anesthetics in financial contexts]. PMID- 11475634 TI - [Perspectives of anesthesia with xenon]. PMID- 11475635 TI - [Do we still need laughing gas?]. PMID- 11475636 TI - [Conus cauda syndrome after spinal anesthesia]. AB - Nowadays spinal cord anesthesia is, when indicated, a standardized and acknowledged medical procedure with a low complication rate. This is also true of neurological after-effects, like spinal paralysis, which are greatly feared, especially by medical laymen. In the following case report we describe a charge against anesthesiologists, who were accused of causing a conus-cauda syndrome after spinal cord anesthesia. Neurological examinations led to the diagnosis of an anesthesia-independent Tethered-Cord-Syndrome (TCS). In most cases the patients' fear can be alleviated by clear explanations about the method. The indication for the method of anesthesia is also based on a thorough body examination. If the documentation of the treatment is faulty or incomplete it can be used against the physician facing a lawsuit more easily. The aim of this case report is to remind anesthesiologists of the rare TCS and to emphasise again the importance of using standardized procedures and correct documentation, which can be very helpful for the physician facing unjustified claims in case of medical litigation. PMID- 11475637 TI - [Bilateral monitoring of cerebral oxygen saturation in aortic arch surgery]. AB - Cerebral ischemia with consecutive neurological damage is a typical complication in aortic arch surgery. Therefore, intraoperative neuromonitoring is of increasing interest. This paper describes the role of bilateral near-infrared spectroscopy (NIRS) in detecting cerebral ischemia. In the case of a patient with acute aortic dissection (Daily, type A), an unexpected sudden reduction of perfusion of the right carotid artery could easily be detected with the help of two instruments INVOS 3100A. The decrease in the saturation values from 74% to 54% correlated well with the drop of blood pressure in the right radial artery. Clamping of the aorta with interruption of the blood flow into the innominate artery through the false lumen was the reason for the low cerebral perfusion. After repositioning the aortic clamp NIRS in combination with invasive blood pressure monitoring sufficiently allowed to control the further course of cerebral perfusion. The NIRS is a non-invasive, easy to handle, and easy to interpret method for intraoperative monitoring of the neurological status. Therefore in our opinion it has got some relevant advantages in contrast to other neuromonitoring methods in aortic arch surgery. PMID- 11475638 TI - [Report on the 28th Neonatal and Infant Respiratory Symposium in Lake Tahoe, Nevada, February 28 to March 3 2001]. PMID- 11475639 TI - [Differential diagnosis of leg ulcers]. AB - Over the last years the treatment of chronic leg ulcers has developed from a merely symptomatic to a differentiated therapy. Although more than 90% of the ulcers are of vascular origin, many other causes are possible. A survey of the various triggers of chronic ulcers of the lower limb is given. Vascular, hematological, neuropathic, infectious, malignant and chemico-physical causes are discussed. New findings regarding hemostatic defects as the basis of thrombophilia and ulcers are presented. PMID- 11475640 TI - [Dermatological prescription law. Guidelines for dermatological prescriptions]. PMID- 11475641 TI - [Perioperative antibiotic therapy in dermatology. Guidelines of the Commission for Quality Assurance of the German Dermatology Society and the Professional Association of Surgical and Oncological Dermatology]. PMID- 11475642 TI - [Incidence of occupation-related skin diseases in skin-exposure occupational groups]. AB - BACKGROUND AND OBJECTIVE: Occupational skin diseases (OSD) rank at the top of occupational diseases in Germany. We studied the cases with initial medical report of an OSD reported to our Register of OSD in North Bavaria (BKH-N). PATIENTS/METHODS: From 1990-99 5.285 cases were closed and recorded. Since the number of employees in the different occupations was known, a population-based study was performed to investigate incidences and demographic characteristics in occupations classified into 24 groups. RESULTS: In the occupational groups in 3.097 cases a work-related skin disease was stated. The estimated overall incidence was 6.7 cases per 10,000 workers per year. The highest incidence was in hairdressers (97.4), bakers (33.2), and florists (23.9), while the largest number of cases was in hairdressers (856), health services (481), and metal-surface workers (260). The median age of all cases was 25 years. In 1.611 cases (52%) a delayed-type sensitisation with occupational relevance was perceived. CONCLUSIONS: The descriptive data of the BKH-N highlight occupational groups being at risk for OSD. The results assist in working out prevention measures aiming on lowering the incidence of OSD. PMID- 11475643 TI - [German instrument for the assessment of quality of life in skin diseases (DIELH). Internal consistency, reliability, convergent and discriminant validity and responsiveness]. AB - BACKGROUND AND OBJECTIVE: Performance and psychometric testing of the German Instrument for the Assessment of Quality of Life in Skin Diseases (DIELH). PATIENTS/METHODS: The instrument consisting of 36 questions (maximum skore 180, 7 domains) was used in 836 outpatients and quality assessments (validity, reliability, responsiveness) were performed. RESULTS: The total skore had a mean of 54 pts. (STD 31, range 0-157) and was normally distributed. Women reported a higher impact on quality of life (median 56 vs. 49, p < 0.001). In terms of discriminant validity significant differences between diagnostic groups were observed with high scorings for patients with eczema, psoriasis, urticaria, and acne (p < 0.001). Tests for internal consistency (Cronbach's alpha 0.71-0.92) and reliability (r = 0.73-0.86) revealed valid results. The DIELH correlated well with the DLQI (r = 0.78, p < 0.001) and moderately with 3 domains of the SF 36 (r = 0.25-0.30, p < 0.01). Changes in the quality of life after 4 weeks were consistent with the corresponding changes of the skin condition and partially significant. CONCLUSIONS: In the use within a large patient sample the DIELH proved to be feasible, valid, reliable, and responsive. PMID- 11475644 TI - [Keratosis lichenoides chronica following trauma. A case report and update of the last literature review]. AB - Since the original description by M. Kaposi in 1895, 40 cases of keratosis lichenoides chronica have been published. Most have been reports of single cases. The clinical appearance is characterized by lichen planus-like papules, linear keratoses, and psoriasis-like plaques. While the etiology is still unknown, an association with a variety of diseases including hepatitis, tuberculosis, glomerulonephritis, and lymphoproliferative diseases has been reported. We present a new case which may have been triggered by trauma or carbamazepine. PMID- 11475645 TI - [Painful bruising syndrome]. AB - A 54-year old anorectic patient with painful bruising syndrome (Gardner-Diamond syndrome) suffered from various gastrointestinal and psychologic complaints. The episodes of painful bruising could be provoked following intradermal injection of washed red blood cells within 96 h. The pathogenesis of this rare entity is more likely due to a psychiatric alteration rather than to the immunological mechanisms, as had been postulated two decades ago. Nevertheless, the Gardner Diamond-syndrome should be included in the differential diagnosis of ecchymotic bleeding. PMID- 11475646 TI - [Bleomycin prick in therapy-resistant verruca vulgaris]. AB - Common warts can be highly resistant to conventional treatments such as cryotherapy, laser surgery, salicylic acid and mechanical removal. Intralesional bleomycin has been described as an effective treatment for common warts since 1970. We treated a 27-year-old woman with Sharp's syndrome who was on immunosuppressive drugs and had suffered for nine years from therapy-resistant warts on palms and soles. A bleomycin-prick technique using a bifurcated needle to introduce bleomycin sulfate (1 mg/ml sterile saline solution) into the warts resulted in almost complete remission after 16 treatment sessions. PMID- 11475647 TI - [Reticular pigmented anomaly of the flexures. Dowling-Degos disease of the intertrigo type in association with acne inversa]. AB - Reticulated pigmented anomaly of the flexures (Dowling-Degos disease) is a rare autosomal-dominant genodermatosis with variable penetrance that is characterized by reticulated hyperpigmentation of the flexures. Acne inversa has been previously described in the literature under several synonyms, such as acne triad, acne tetrad and hidradenitis suppurativa. Acne inversa is an inflammation affecting the pilosebaceous units of the flexures. We report a case of Dowling Degos disease in association with acne inversa. The association of these diseases has been described several times and it appears not to be coincidental. Pathogenetic mechanisms leading to this coincidence are discussed. PMID- 11475648 TI - [Extensive ulcerated basalioma. Irradiation with ionizing radiation after electrocautery debridement]. AB - Soft X-Ray (Dermopan) is an effective and gentle treatment for large basal cell carcinomas of the face. It is an useful alternative to extensive and often cosmetically unsatisfactory surgery, especially in large basal cell carcinomas covering an area of more than 4 cm2. A 79-year-old man presented with a 4,5-year history of an extensive basal cell carcinoma on the right temple, which was treated by soft x-ray after removal of excessive tumor tissue with electrocautery. A total dosage of 48 Gy was employed. PMID- 11475649 TI - [Clinical and histopathological characteristics of early Leser-Trelat syndrome]. AB - Leser-Trelat-syndrome is characterized as the eruptive appearance of multiple seborrheic keratoses in association with underlying malignant disease. A 73 year old female patient with a metastatic adenocarcinoma of the colon presented with this paraneoplastic change. The rapid appearance of solitary seborrheic keratoses with associated inflammation, seen clinically and histologically, may be an early sign of Leser-Trelat-syndrome. The recognition of this inflammatory component as an early sign may contribute to the prompt diagnosis of this paraneoplasia, even before the eruption of numerous seborrheic keratoses. PMID- 11475650 TI - [The vacuum sealing technique. A new procedure to cover soft tissue defects after resection of leiomyosarcoma]. AB - The vacuum sealing technique is a new, simple to use procedure for traumatic and chronic soft tissue defects, burns and soft tissue infections. We used this technique for the first time for temporary closure after the resection of deep infiltrating leiomyosarcoma. Radical excision of the tumor with partial resection of the forearm flexor muscles and safety margin of 3 cm led to a large skin defect (12 x 16 cm). For temporary closure of this defect we used the vacuum sealing technique for 7 days. Because of histologically proven incomplete resection of the tumor, we did a second excision and used the vacuum sealing technique a second time for 7 days. After this time period the granulation of the wound ground was excellent, and we closed the defect by split skin transplantation from the thigh. Bacterial infection or other complications were not seen during use of vacuum sealing technique. The vacuum sealing technique induced optimal local conditions for skin grafting: it facilitated granulation tissue production and maintained a clean wound bed without the necessity of changing wound dressing before skin grafting. This technique is painless and does not impair mobility. PMID- 11475651 TI - [Black nails caused by Proteus mirabilis]. AB - Black nails caused by Proteus mirabilis were seen in a motor mechanic and in a petrol pump attendant. Proteus mirabilis is a gram-negative bacillus that generates hydrogen sulfide. This compound reacts with traces of metals in the nail plate such as zinc, nickel, cobalt, iron, manganese, tin, copper and lead. Metal sulfides blacken the nail plate. The protracted course of the discoloration over months corresponds to the slow reactions of metals with hydrogen sulfide. The disappearance of the blackening after topical treatment with chinosol, tincture of iodine and chloramphenicol solution supports the etiologic connection between black nails and Proteus mirabilis. Wet and dirty work encourages the colonisation of Proteus mirabilis between nail fold and nail plate. PMID- 11475652 TI - [Wax model collection of the Department of Dermatology, University of Munster]. AB - Between the 17th century and the middle of the 20th century, wax models were widely used for training in anatomy and other medical fields, especially dermatology. Thereafter, photographic techniques led to the replacement and subsequent dissolution of many European wax model collections. Unfortunately many wax models lose their original colors over time. Since only few mouleurs (wax artists) are still active in Europe conserving wax models, the "dying of moulages" cannot be stopped. Consequently, the existing wax models are of medical and historical interest since they reflect the development of our speciality. The Department of Dermatology in Munster is in the possession of 121 wax models, which are presented in this article. PMID- 11475653 TI - [A medical student discovers onychomycosis. Comment on the article by N. Haas and H. Sperl: Hautarzt (2001 52:64-67]. PMID- 11475654 TI - [Painful swelling of the foot of a 16-year-old African. Eumycetoma (Madura foot)]. PMID- 11475655 TI - [Nuclear factor kappa B (NF-kappa B). I. Function and regulation]. PMID- 11475656 TI - Opening the black box: why we need a PBL talkbank database. AB - BACKGROUND: Interest runs high these days in developing "evidence-based" reviews to provide guidelines for instructional practice. However, we lack careful documentation of the ways in which the practices of problem-based learning (PBL) vary across groups and across implementations. A necessary starting point for developing any sweeping conclusions about the efficacy of PBL as an instructional innovation, therefore, is that we begin to become more articulate about what it is that people do when they say they are doing PBL. SUMMARY: A proposal is offered for a new initiative in medical education research, one focused on documenting the range of practices employed in different implementations of PBL. A vital facet of this initiative would be the development of a shared corpus of video recordings referred to here as the "PBL TalkBank database." CONCLUSIONS: We propose that medical educators adopt the tradition employed in linguistics and communication studies of creating shared data corpora. The corpus in this case would consist of recordings, transcripts, and research notes documenting PBL practices in different PBL curricula. Preliminary work has been undertaken to develop such a database, and we invite the participation of other researchers. PMID- 11475657 TI - Promoting collaborative teaching in clinical education. AB - BACKGROUND: Although the practice of medicine is increasingly a multidisciplinary effort, clinical teaching of medical students is accomplished primarily within a departmental structure. PURPOSE: The purpose of this study was to identify subject matter within the clinical curriculum that could serve as focus for multidisciplinary teaching. METHODS: A questionnaire was sent to 13 clerkship directors (representing required clerkships) at a large Midwestern medical school in which they were asked to rate a list of 631 patient problems as critical (primary), important (secondary), or "nice to know," relevant to their respective clerkship objectives. RESULTS: All clerkship directors completed the questionnaire. There were 523 items that were considered primary, and over 90% of these items were listed as either primary or secondary in more than 1 clerkship. Twelve topics were considered primary or secondary by at least 5 clerkship directors. Four clerkship directors identified 43 patient problems, and 3 clerkship directors identified 92 topics as primary or secondary clerkship objectives. CONCLUSIONS: In this study, listing of patient problems across clerkships demonstrated significant overlap of the clinical curriculum, suggesting multiple opportunities for faculty collaboration in clinical education. PMID- 11475658 TI - What's the use of faculty development? Program evaluation using retrospective self-assessments and independent performance ratings. AB - BACKGROUND: The assessment of the effectiveness of faculty development programs is increasingly important in medical schools and academic medical centers but is difficult to accomplish. PURPOSE: We investigated the usefulness of retrospective self-assessments by program participants in combination with independent ratings of teaching performance by their trainees. METHODS: We used a single sample, prepost intervention design using multiple measures. Our assessment instruments were based on our institution's accepted teaching competencies. We measured participants' self-assessments of their teaching competencies before the program and their retrospective self-assessed improvements in these competencies after the program. We also used independent ratings of the participants' teaching competencies before and after their involvement in the program, as rated by their own trainees (fellows, residents, and medical students). Selected teaching competencies comprised the intended learning outcomes of the faculty development program. RESULTS: Participants' preprogram self-assessments showed that the program was appropriately matched to several topics identified as needy, but also included topics that participants did not identify as needs. The retrospective self-assessments showed improvements in teaching skills that previously were identified as needs, as well as those in which participants originally felt quite competent. The independent ratings by trainees showed overall positive improvements (some significantly). The retrospective self-assessed improvements correlated positively with the independent ratings by their trainees (p < .01). CONCLUSIONS: This evaluation strategy showed that the faculty development program improved the teaching competencies of the participants. Both the program participants' retrospective self-assessments and the independent ratings by their trainees showed postprogram improvements and were positively intercorrelated. The use of these multiple measures is a viable approach to evaluate the impact of a faculty development program. Potentially either approach could be used, but in combination, they provide a feasible, valid, and reliable evaluation. PMID- 11475659 TI - Error patterns of 3rd-year medical students on the cardiovascular physical examination. AB - BACKGROUND: Recent research documents widespread deficits in the physical examination skills of practicing physicians. PURPOSE: This study explored physical examination skills of 3rd-year medical students after completion of a course in physical diagnosis. METHODS: Standardized patient physical examination checklist data were analyzed for a cohort of 2,038 medical students for a patient presenting with classic signs and symptoms of an acute myocardial infarction. A follow-up paper case and survey explored reasons underlying omissions. RESULTS: Students systematically omitted 3 of 10 component maneuvers critical to the evaluation of a patient with shortness of breath and chest pain. The same pattern of omissions was observed across 8 medical schools and over 2 successive years. The paper case follow-up study ruled out time constraints and performance anxiety as the cause. Survey data revealed that students may omit a maneuver due to inability to recall pertinence (blood pressure in both arms) or difficulty discriminating findings (heart sounds at different locations), or because of inadequate technical mastery (percussion of the lungs). CONCLUSIONS: These data suggest fundamental inadequacies in the current paradigm for teaching physical examination skills. Standardized patient checklist data can provide an informative window into the efficacy of teaching practices. PMID- 11475660 TI - Medical students' experience with psychiatric diagnoses in a family medicine clerkship. AB - BACKGROUND: Many medical schools require a family medicine clerkship, yet little is known about the quantity and diversity of the diagnoses encountered by the students. PURPOSE: This study examines patients encountered with psychiatric diagnoses using quantitative data collected by students in a family practice clerkship. METHODS: Over a 2-year period, 445 students completed 3,320 patient encounter forms for patients with a psychiatric diagnosis, noting their comfort level and responsibilities. RESULTS: The patients' diagnoses reflect those seen in a typical family practice. Of the 71,869 presenting diagnoses, 3,548 were for a psychiatric condition, most commonly depression (37.1%) and neuroses (28.0%). Students reported a high level of comfort in diagnosing and treating patients with a psychiatric disorder. The students routinely discussed these cases with their preceptors. CONCLUSIONS: By using a relatively simple computerized database, many curricular issues can be identified. For example, analysis of the database shows that the clerkship provides students with substantial practice in taking patient histories and performing initial patient examinations in patients presenting with a psychiatric problem. However, students infrequently provided patient education and counseling to patients with psychiatric disorders. Specific psychiatric diagnoses reflecting limited experience and lower levels of perceived competence include attention deficit disorder and senile and presenile organic psychotic disorders. PMID- 11475661 TI - Reinforcement of occupational history taking: a success story. AB - BACKGROUND: This article describes the results of a retrospective study of 3 classes of medical students who participated in a targeted occupational and environmental health curriculum at the University of Connecticut School of Medicine. PURPOSE: We wanted to determine if targeted focused curricular interventions which integrated occupational and environmental health principles into routine history taking would result in increased scores on the number of questions posed during the Clinical Skills Assessment Program in the 4th year. METHODS: We analyzed Clinical Skills Assessment Program questions for 3 graduating medical school classes from 1997 to 1999. RESULTS: It appears that intense, focused training may increase the occupational and environmental questions which students ask. By revisiting the components of the history during the 3rd year, the final assessment of 4th-year students substantially and significantly increased. CONCLUSIONS: Those who wish to stem the decline in history-taking skills as students enter their clinical years should consider reinforcing these skills using structured programs and practice in areas of the history that are traditionally neglected but recognized as essential in gathering comprehensive data on patients. PMID- 11475662 TI - Oncology thinking cap: scaffolded use of a simulation to learn clinical trial design. AB - BACKGROUND: Physicians often are called on to participate in and interpret clinical trials, but their training in this area may not provide them with the inquiry skills that are needed. Simulations have the potential to be a promising tool for helping medical students learn the skills involved in clinical trial design. However, simulations may be complex and require additional scaffolding to support learning. DESCRIPTION: The goal of this study was to teach aspects of cancer clinical trial design through the scaffolded use of a simulation, the Oncology Thinking Cap. The software-based scaffolding provided guidance in designing the trial. Subsequently, the simulation allowed students to run the designed trial, which produces detailed patient histories. This feedback then could be used to redesign the trial. EVALUATION: Twenty-four 4th-year medical students were asked to design a clinical trial in advance, on paper, to test a new anticancer drug. Student groups then designed and simulated running the clinical trial assisted by the software environment. Instructional effectiveness was measured using a pretest-posttest design that included having students (a) write a group research proposal and (b) individually critique a flawed proposal. At the group level (N = 6 groups), students demonstrated a 34% increase in the number of elements of a clinical trial that they included in their research proposals. At the individual level (N = 24), students improved by 48% in their critiques of flawed proposals. CONCLUSIONS: Scaffolding embedded in the simulator is a promising approach to helping students learn about clinical trial design. PMID- 11475663 TI - Duke's 3rd year: a 35-year retrospective. AB - PURPOSE: Our purpose was to review and evaluate Duke University School of Medicine's 3rd year. SUMMARY: Duke University School of Medicine instituted a dramatic change in its curriculum 35 years ago by restructuring the 1st, 2nd, and 4th years of medical school so that the entire 3rd year could be devoted to an elective research experience or independent study of the basic sciences. Today this "new curriculum" permits students to work in basic science laboratories or with clinical investigators, work toward a 2nd degree, and participate in basic science courses beyond the core curriculum. Students were surveyed in 1996 and 1998 about their 3rd-year experience. Results showed that 67% and 73%, respectively, believed the 3rd year was very helpful in facilitating their understanding and appreciation of biomedical and clinical research. Students especially valued the experience and availability of their preceptors and the range of opportunities available during the 3rd year. The most frequent critiques by students were the lack of opportunities to pursue clinical interests, difficulty integrating the 3rd year into the remainder of the curriculum, and a tendency to place too much emphasis on coupling the 3rd-year experience with scholarships and residency plans rather than on a specific investigative interest. CONCLUSIONS: Overall, both faculty and students value the 3rd year as a unique positive experience. Duke faculty have noted, however, that no other medical schools have attempted to replicate the Duke experience. PMID- 11475664 TI - Locally produced specific cytomegalovirus immunoglobulin for intramuscular use as a therapy in renal allograft recipients. AB - Immunomodulator, i.e. specific hyperimmune anticytomegalovirus immunoglobulin for intramuscular administration, produced in 1999 with the aim of prevention of CMVI, and the development of the disease, was for the first time applied in kidney transplant recipients in January 2000, in the Center for kidney transplantation at the Military Medical Academy. Therapy was administered in four cytomegalovirus (CMV)--seronegative kidney recipients from CMV-seropositive donors--the combination that in the majority of cases lead to the development of CMVI/disease resulting in transplant rejection. Patients received 0.2-0.3 ml/kg of cytomegalovirus immunoglobulin (CMVIG) 6 hours before the transplantation, and subsequently the same dose during the following 5 weeks. Simultaneously, they received ganciclovir in therapeutic doses adjusted according to creatine clearance during the first three post-transplantation months (2 weeks parenterally, the rest orally). Kidney transplant recipients tolerated well i.m. applied CMVIG without any adverse effects. Test result obtained from the Paul Erlich Institute, Germany in 1999 spoke in favor of the quality of the first national CMVIG preparation. PMID- 11475665 TI - [Vitreoretinal surgery in war injuries of the eye]. AB - Results of vitreoretinal surgeries of 647 severe penetrating injuries of the eye, sustained during the war operations in the territory of former Yugoslavia, in the period from 01.07.1991. to 31.12.1998. were analyzed in the paper. A total of 558 pars plana vitrectomies, 459 intrabulbar foreign bodies' extractions and 360 surgeries of traumatic retinal detachments (89 in the conventional way) were performed at the Clinic of Ophthalmology--Department for Vitreoretinal Surgery of the Military Medical Academy. Certain innovations in the surgical treatment of the severely injured eye with the damage of posterior eye segment, as well as the lens, were presented. In those cases, (simultaneously with pars plana vitrectomy, and extraction of retained intrabulbar foreign body), the primary intraocular lens (IOL) implantation was also performed, particularly posterior chamber IOL implantation following lensectomy, as well as phacoemulsification. PMID- 11475666 TI - [Dislocation of the knee joint]. AB - Dislocations are the rarest but the most severe injuries of the knee joint. Six patients with traumatic knee joint dislocation were treated at the Clinic for Traumatology and Orthopedics of the Military Medical Academy in the period January 1989-November 1998. Popliteal artery was injured in 3 patients. The injuries required reconstruction and the treatment in 1 patient ended with upper leg amputation. In one case the diagnosis was irreducible lateral dislocation. Surgical reparation--reconstruction of ligamentary apparatus was performed in 4 patients. Concerning the knee joint stability satisfactory results were achieved. Knee joint dislocations are very severe injuries requiring thorough diagnosis of the possible injury of neurovascular bundle and surgical treatment. PMID- 11475667 TI - [Assessment of embolism risk in deep vein thrombosis in various locations using radionuclide phlebography and perfusion scintigraphy of the lungs]. AB - With the aim of the estimation of embolism at certain locations of deep vein thrombosis (DVT) 3 groups comprising 22 patients were investigated in whom isolated thrombosis in one of the deep veins segments: lower leg, upper leg, and pelvis was determined by radionuclide phlebography (RNF) and contrast phlebography. Afterwards, all necessary investigations relevant for pulmonary thromboembolism (PTE), including perfusion scintigraphy of the lungs (PSL) were performed. Out of all patients with DTV, 17% were with pulmonary changes confirming PTE. Out of patients with DVT, 59% were with the changes in the upper leg segment, and 66% of patients were with changes in pelvic segment, respectively. PMID- 11475668 TI - [Septic encephalopathy--prognostic value of the intensity of consciousness disorder to the outcome of sepsis]. AB - Septic encephalopathy (SE) is a common term indicating the development of signs of progressing cerebral dysfunction and is associated with the presence of microorganisms and their toxins in the blood. Aim of this investigation was to analyze the frequency of this complication considering the consciousness disorders in quantitative sense and prognosis of the survival in patients with SE. The investigation comprised patients (n = 54) with positive hemoculture and signs of septic syndrome by the accepted criteria (fever, clinical signs of infection, respiratory frequency, heart rate, plasma lactate, oliguria). Patients with confirmed cerebral injury, hemorrhage or cerebral ischemia were excluded from the study. Lumbar punction and CT-scan of the brain were performed in all patients in order to exclude visible lesions of cerebral parenchyma and eventual presence of cerebral nervous system (CNS) infection as the causes of sepsis. Results of the investigation demonstrated that in 30 (55%) of patients existed mild consciousness disorder at the level of somnolescence, in 18 (33%) consciousness disorder at the level of sopor and in 6 (11%) consciousness disorder at the level of deep coma. Level of consciousness disorder was in positive correlation with the outcome of sepsis syndrome, which was additionally confirmed by the fact that only in the group of patients with deep coma lethal outcome was observed in 3 cases (50% of this subgroup) regardless of intensive antibiotic, metabolically active and symptomatic therapy. It can be concluded that SE syndrome has a favorable prognosis if macroscopic lesion and dissemination of microorganisms in CNS are not present, and simultaneously it represents changes in metabolic-electrolytic state with early presentation of consciousness disorders that represent clinically significant indicator for sepsis syndrome outcome. PMID- 11475669 TI - [Use of mycophenolate mofetil in patients with a transplanted kidney and cyclosporin nephrotoxicity]. AB - Cyclosporine (CsA) nephrotoxicity is an important problem in renal transplant recipients, which can influence long-term graft survival. The safety of conversion from CsA to azathioprine (AZA) remains controversial and can result in higher incidence of acute rejection. Mycophenolate mofetil (MMF) is a new immunosuppressive agent superior to AZA in the prevention of acute rejection. Five patients with cyclosporine nephrotoxicity were converted from CsA/AZA/prednisolon to MMF/prednisolon protocol. All patients had low immunological risk and 4 out of 5 patients received antithymocyte globulin before conversion as the induction therapy or as the treatment for acute rejection. Mean follow-up after conversion was 16.8 months (range 4-32 months). No patient experienced acute rejection during follow-up period. The mean serum creatinine concentration decreased from 219 +/- 44.18 (range 168-280) to 122.6 +/- 48.02 mumol/l (range 72-187 mumol/l) (p = 0.002). Arterial hypertension improved after CsA withdrawal in 20% of patients. We have concluded that, in selected patients with cyclosporine nephrotoxicity, CsA withdrawal with concomitant use of MMF is safe and effective in the improvement of graft function and arterial hypertension. PMID- 11475670 TI - [Acute post-streptococcal glomerulonephritis in adults--corticosteroid therapy, yes or no?]. AB - The aim of this study was to investigate the influence of the corticosteroid therapy on the development and prognosis of proststreptococcal glomerulonephritis (PSGN) in the period of five years. The investigation included a group of 54 patients who were in the acute phase of poststreptococcal glomerulonephritis (PSGN) when they were 18-22 years old. Corticosteroids were administered per os with the starting dose of 0.5 mg/kg bm, and the dose was subsequently decreased by 5 mg every 10 days to 20 mg/per day within 8 weeks of therapy. We have treated ten patients and all were with severe clinical manifestations of the disease (acute renal failure, oliguria, edemas, hypertension) and severe histopathologic (HP) finding of kidney tissue on the initial biopsy. On the basis of clinical parameters, HP changes on the repeated biopsy of the kidney and five years follow up, we have concluded that the remission of the disease was achieved in patients who had received corticosteroid therapy. Clinical findings were confirmed by PH findings of repeated biopsies with less expressed residues of the disease. Corticosteroid therapy should be administered in adult patients who are in the acute phase of the disease, with clinical and HP severe form of PSGN, since the risk for the further progress of more serious sequelae of the disease is significantly decreased. PMID- 11475671 TI - [Traumatic amnesia: neuropsychological analysis and clinical symptoms]. PMID- 11475672 TI - [Cytokines--physico-chemical properties and major biological characteristics]. PMID- 11475673 TI - [Electroconvulsive therapy--current approach]. PMID- 11475674 TI - [Current problems in medical ethics]. PMID- 11475675 TI - [Successful surgical removal of a cardiac myxoma from the left ventricular outflow tract]. AB - The precondition for successful, exclusively surgical treatment in the extracorporeal circulation, is the precise diagnosis of heart myxoma, particularly in rare locations such as the left ventricle. We present a case of myxoma in the outflow tract of the left ventricle as the exceptionally rare location, successfully diagnosed and surgically treated at the Clinic for Cardiac and Thoracic Surgery of the Military Medical Academy. A female patient, aged 46 years, was sent from another hospital with misdiagnosis of idiopathic hypertrophic subaortic stenosis. The patient was successfully operated after transthoracic and transesophageal echocardiography as the main diagnostic procedures. Myxoma that completely obstructed the aortic opening if pulled, was completely removed through aortic valve in the extracorporeal circulation. Its pedicle was arising from the ventricular side of the great mitral cusp. Postoperative course was uneventful and the patient was released from the hospital on the tenth postoperative day. PMID- 11475676 TI - [Neuroleptic malignant syndrome with rhabdomyolysis and muscle rupture]. AB - We report a case of neuroleptic malignant syndrome (NMS) development after the ingestion of 960 mg of haloperidol in 18-year old male on the previous long-term neuroleptic treatment. Beside the severe rigidity and laboratory signs of rhabdomyolysis, firm swelling of gluteal and femoral areas, accompanied by pain and difficulty in the extension of the right leg, was noticed. The ultrasonography of the affected limb revealed partial rupture of quadriceps muscle. Since no signs of trauma or any other causes of muscle rupture were observed, we concluded that it developed due to the severe hypertonia and rhabdomyolysis in NMS. As there were no such cases previously reported, we wished to point out the possibility of muscle rupture development in NMS and propose the adequate diagnostic procedures and treatment. PMID- 11475677 TI - [Primary liposarcoma of the pericardium]. AB - A case of primary liposarcoma of the pericardium, an exceptionally rare localization, has been presented. The tumor localized in the region of the left ventricle and connected to the visceral wall of the pericardium was diagnosed in a 38-year old male. Histological, histochemical and immunochemical findings confirmed the diagnosis of the liposarcoma of the pericardium. PMID- 11475678 TI - [Nobel Prize winners in surgery--part 3]. PMID- 11475679 TI - [Nutritional and metabolic status and dietetic evaluation in institutionalized elderly patients with non-insulin-dependent diabetes mellitus]. AB - OBJECTIVE: To evaluate nutritional status and blood glucose level according to type of diet followed by elderly (> 65 years old) institutionalised patients with non insulin dependent diabetes mellitus (NIDDM). METHODS: A data collection questionnaire was administered to the staff of 80 Spanish geriatric facilities participating in this one-day cross-sectional study. Data collected included: age, gender, history of previous glucose control, type of antidiabetic treatment, body mass index (BMI), nutritional risk index (NRI), type of diet with patient's preference, nutritional status evaluation and biochemical parameters. RESULTS: Data from a total of 486 institutionalised elderly people (66.5% female) were collected with a mean +/- SD age of 80.8 +/- 7.5. Had a diabetic diet 93.3% of patients with no gender differences 55.7% were on oral antidiabetic treatment (53.9% females and 58.9% males, ns) and 24.7% on insulin therapy (24.5% females and 26.4% males, ns). Physical activity was presented in 86% of people with no gender differences. Exercise was practiced by people younger, 78.8 +/- 7.9 years, versus walking, 80.4 +/- 7.6 years or resting 82.1 +/- 7.3 years, p < 0.05. Both nutritional indexes, BMI and NRI, were equivalent in men and in women; 28.5 +/- 10.7 vs 28.1 +/- 7.6 and 103.5 +/- 12.9 vs 104.1 +/- 11.4, respectively. History glucose control was good in 68.8% of the sample with a trend towards significance between sexes (62.7% males, 71.7% females, p = 0.07) and significant differences according to type of diet; 70.7% diabetic versus 37.0% free diet, p = 0.0006). Eating behaviors (appetite and meal completeness) were similar in both males and females and in diabetic or free diet groups. Also, no differences were observed in patient preferences about diet according to type of diet (71% of diabetic patients always or almost always liked their food versus 82% of free diet patients) and either nutritional status (9.3% vs 15.6% in status I, 10.9% vs 12.5% in level II and 79.9% vs 71.9% in status III, respectively). The NRI, cholesterol, albumin and triglyceride levels were significantly higher in patients with high fat rate supplemented dietary intake than other supplemented diets. CONCLUSIONS: A high proportion of patients on a diabetic diet was observed in this study with no gender differences. Elderly NIDDM institutionalised patients on a diabetic diet showed a greater good history glucose control than patients on a free diet. Diabetic diet preference by these subjects is similar than free diet. A better nutritional risk control was shown in patients with high fat rate supplemented diet. PMID- 11475680 TI - [Nutrition in the surgical patient: immunonutrition]. AB - Malnutrition plays an important role in the rate of postoperative complications that interfere with our surgical activity, impairing immune response mechanisms; synthesis and regeneration processes are damaged and the fight against infection is altered. Preoperative and postoperative administration of diets enriched with diverse substances included under the name of "immunonutrients" reduces the rate of complications. Our knowledge about the mechanisms of action of each immunonutrient is increasing as well as the fact that the combined action of these substances improves the immune system and protects the organism against the negative effect of the systemic inflammatory response syndrome. PMID- 11475681 TI - [Protective effect of monounsaturated and polyunsaturated fatty acids on the development of cardiovascular disease]. AB - Cardiovascular disease has a multifactorial aetiology, as is illustrated by the existence of numerous risk indicators, many of which can be influenced by dietary means. In this article, the effects of unsaturated fatty acids on cardiovascular disease are reviewed, with special emphasis on the modifications of the lipoprotein profile and the mechanism by which fatty acids may affect the immune response on the development of the atherosclerotic lesion. Atherosclerosis occurs fundamentally in three stages: dysfunction of the vascular endothelium, fatty streak and fibrous cap formation. Each of the three stages is regulated by the action of vasoactive molecules, growth factors and cytokines, mediators of the immune response. Dietary lipid quality can affect the lipoprotein metabolism, altering their concentrations in the blood, permitting a greater or lesser recruitment of them in the artery wall. The replacement of dietary saturated fat by mono- or polyunsaturated fats significantly lowers the plasma-cholesterol and LDL-cholesterol levels. Likewise, an enriched monounsaturated fatty acid diet prevents LDL oxidative modifications more than an enriched polyunsaturated diet, and the oxidation of LDL in patients with peripheral vascular disease mediated by n-3 fatty acids can be reduced by the simultaneous consumption of olive oil. However, strong controversy surrounds the effect of the different unsaturated fatty acids. The type of dietary fat can directly or indirectly influence some of the mediating factors of the immune response; n-3 fatty acids have powerful antiinflammatory properties. Dietary fatty acids strongly determine the susceptibility of lipoproteins to oxidation, which also has an impact on the activation of molecules of adhesion and other inflammatory factors. Moreover, several works have demonstrated a direct effect of fatty acids on the genetic expression of many of those factors. Finally, certain aspects of blood platelet function, blood coagulability, and fibrinolytic activity associated with cardiovascular risk, are modulated by dietary fatty acids; n-3 fatty acids strongly inhibits platelet aggregation and activate thrombolytic processes. PMID- 11475682 TI - [Dietary and biochemical riboflavin status in a cohort of young people in the community of Madrid]. AB - Riboflavin status was measured in 228 young people (101 women and 127 men) between 18 and 35 years of age, resident in the Comunidad Autonoma de Madrid. A dietetic study was undertaken using a three day food record (which included one non-working day). Also, biochemical levels of riboflavin were investigated via the determination of the activation coefficient of erythrocyte glutathione reductase (alpha-EGR). Although the mean intake of riboflavin was acceptable (107.1 +/- 29.1 of recommended intakes), 46.9 of subjects showed intakes below recommended. At the biochemical level, 14.9% of subjects showed a moderate risk of vitamin B2 deficiency (alpha-EGR: 1.2-1.4) whilst 1.3% were at high risk (alpha-EGR > 1.4). Male subjects showed higher alpha-EGR levels than did females, which coincides with their greater intake of vitamin B2. Milk products provided some 40.4% of the vitamin, meats 19.9%, cereals 8%, vegetables 7.2%, eggs 6.3%. Other foods provided lesser amounts. Therefore, although riboflavin is available across the food groups, it Would seem that milk products are its most important source. Indeed, subjects with high milk product intakes (more than one ration per day) showed the highest riboflavin levels and the most adequate alpha-EGR levels. This study shows that riboflavin status can be improved, especially amongst those with low milk product intakes. An increase in the consumption of these foods to recommended levels (2-3 rations per day) might improve this situation at both dietetic and biochemical levels. PMID- 11475683 TI - [Evaluation of nutritional risk in ambulatory elderly patients]. AB - OBJECTIVE: The purpose of this study was to evaluate the nutritional risk in ambulatory elderly people (> or = 65 years old) and determine influential factors. METHODS: A questionnaire was provided to 400 Spanish pharmacists to gather data from ambulatory elderly people in a one-visit prospective study. Data collected included: age, gender, body mass index, environment and nutritional assessment using the Nutrition Screening Initiative panel, a subjective nutritional evaluation and nutritional therapy. ANOVA and Mann-Whitney tests were used to evaluate differences and correlations. Logistic regression analysis was used to calculate association with risk factors. RESULTS: Data were collected from 1320 elderly subjects (60.0% female, mean +/- SD, age = 75.6 +/- 7.4). Moderate to high nutritional risk was present in 79.1%, without gender differences, and 18.6% were undergoing nutritional therapy of whom 93% consumed a complementary enteral diet. The following nutritional risk factors were identified (Odds ratio -95% confidence interval-): having three or more glasses of beer or wine in men (10.70, -6.11 to 18.70-), eating usually alone for women (1.96, -1.52 to 2.52-) and physical weakness for cooking, shopping or feeding in women (1.7, -1.34 to 2.16-). The assessment of nutritional status with the Nutrition Screening Initiative panel showed a linear significant correlation with age (p < 0.01). There was poor correlation between the objective (panel) and subjective evaluations (kappa index, 0.0822). The environment was not associated with nutritional status, but women showed a poorer subjective nutritional valuation than men (p < 0.001). Although chronic diseases and drug therapy are usually the most frequent risk factors, in our subjects, alcohol consumption was the most frequent risk factor. Poor fitness showed a very close association with a moderate to high nutritional risk. There is a high rate of elderly people at risk and a very low percentage of subjects undergoing nutritional therapy. CONCLUSIONS: The moderate to high nutritional risk rate in ambulatory Spanish elderly persons is higher than expected. Both physical and social issues influence nutritional status. Awareness of the high rate of nutritional risk, should prompt consideration of early, appropriate therapy to prevent malnutrition and deteriorating quality of life. PMID- 11475684 TI - Outcomes assessment an intervention strategy to improve parental compliance to follow-up evaluations after treatment of early childhood caries using general anesthesia in a Medicaid population. AB - Young children from low-income families are at risk for the development of early childhood caries (ECC) that can progress to severe oral disease. Treatment of this condition often requires extensive rehabilitation using general anesthesia in an ambulatory care facility. These children, presenting with neglected oral health, frequently face major obstacles to accessing dental care in a timely manner. Recently, several retrospective studies reported poor follow-up compliance (return for recall evaluations) in children treated for ECC using general anesthesia (GA). The purpose of this study was to provide a prospective analysis of an intervention strategy aimed at improving follow-up compliance in this population. Results of the analysis suggested that the insertion of an additional pre-operative consultation appointment failed to improve significantly compliance to follow-up evaluations or change parental dental health knowledge and preventive practices. Patient variables studied also failed to discriminate influences on predicting compliant behavior. Although retreatment (relapse) was prevalent among those patients who complied with follow-up evaluations, a statistically significant improvement in plaque, gingival, and mutans streptococci scores were demonstrated, following the degree of aggressive restorative treatment typically provided using general anesthesia. These findings are contrary to those reported from conventional restorative therapy and might reflect a beneficial outcome of an aggressive restorative approach. Further investigation is required to identify an intervention strategy that improves follow-up compliance and reduces the costly ravages of dental neglect in young children from low-income families. PMID- 11475685 TI - Treatment planning for space maintenance in the primary and mixed dentition. AB - This manuscript provides the current standard passive space maintenance recommendations and identifies the appropriate space maintainer for premature loss of teeth in the primary dentition, early mixed dentition, and late mixed dentition. PMID- 11475686 TI - Kinetic cavity preparation effects on secondary caries formation around resin restorations: a polarized light microscopic in vitro evaluation. AB - The purpose of this in vitro study was to compare the effect of conventional handpiece and kinetic cavity preparation (KCP, air abrasion) techniques of cavity preparation on caries-like enamel lesion formation. After a fluoride-free prophylaxis, twelve human molars were examined macroscopically to ensure that buccal and lingual surfaces were caries-free. Unfilled cavities were prepared in mesiobuccal (conventional [CU]) and mesiolingual (air abrasion [AU]) enamel surfaces. Cavities were prepared in distobuccal (conventional [CF]) and distolingual (air abrasion [AF]) enamel surfaces and restored with composite resin following placement of a bonding agent. Acid-etching of cavity walls was performed only with the conventionally prepared cavities restored with resin. Air abrasion (KCP) prepared cavities were restored without acid-etching of the cavity walls. Teeth were thermocycled (500 cycles, 5 degrees to 50 degrees C, 500 cycles) and exposed to an artificial caries medium for caries initiation and progression. After caries formation, two longitudinal sections were taken from unfilled and filled cavity preparations and examined by polarized light microscopy for wall lesion presence and mean surface lesion depth. Surface lesion depths were similar among groups after the caries initiation period (CU = 225 microns; AU = 237 microns; CF = 241 microns; AF = 251 [p > .05, ANOVA, DMR]), and progression (CU = 437 microns; AU = 415 microns; CF = 405 microns; AF = 429 um [p > 0.05, ANOVA, DMR]). Extensive wall lesions were present in all CU and AU; while small wedge-shaped wall lesions were significantly (p < .05, ANOVA, DMR) fewer in CF (19 percent & 21 percent) and AF (17 percent & 21 percent) following caries initiation and progression compared with unfilled controls. Resin restorations placed in cavities prepared by air abrasion (KCP) and conventional handpiece techniques provided similar degrees of protection against a secondary caries-like challenge. PMID- 11475687 TI - In vivo rehardening of enamel eroded by a cola drink. AB - There are many concerns about the erosive effects of acidic beverages. In this study, the effects of Pepsi-Cola (pH 2.41) on bovine enamel and the rehardening effect resulting from intraoral exposure of the teeth were determined by microhardness testing (Vicker's Hardness Number). Bovine enamel specimens (VHN 380.00 +/- 12.74) were immersed in 100 ml of Pepsi-Cola for five minutes and subsequently exposed to human intraoral environment through the use of a removable resin plate. Microhardness testing was performed on specimens after one hour, twenty-four hours and forty-eight hours of exposure to an intraoral environment. The microhardness value was significantly (p < 0.05) reduced by the cola beverage (VHN 262.13 +/- 20.34), and significantly (p < 0.05) increased after intraoral exposure for one hour (VHN 299.75 +/- 26.86) and twenty-four hours (VHN 328.00 +/- 18.70). The difference in the microhardness between the twenty-four-hour group and the forty-eight-hour group (VHN 333.50 +/- 15.13) was not significant (p > 0.05). The microhardness value of the forty-eight-hour group was significantly less than the values recorded during the initial pre-study measurements p < 0.05). PMID- 11475688 TI - Remineralizing effects of glass ionomer restorations on adjacent interproximal caries. AB - The purpose of this study was to examine the in vitro remineralization of incipient caries lesions on teeth adjacent interproximally to teeth with Class II glass ionomer cement restorations. Sixty-four extracted molars were selected and 1 x 5 mm artificial caries lesions were created at the interproximal contact point. One hundred micrometer sections were obtained at the caries sites, and polarized-light photomicrographs were obtained. The sections were covered with varnish, leaving only the external section site exposed, and were placed back into the original tooth. In another sixty-four molars, Class II cavities were prepared. Equal numbers of preparations were filled with Fuji IX GP, Vitremer, Ketac-Molar, or Z 250. These sixty-four teeth were mounted to have interproximal contact with the adjacent teeth containing the artificial caries lesions. Specimens were placed in closed environments of artificial saliva for one month. After thirty days, the same sections were photographed again under polarized light, and areas of the lesions were quantitated. Decrease in the size of caries lesions indicated the glass ionomers had significantly greater remineralization effects on adjacent caries than the nonfluoridated composite resin (ANOVA p < 0.05). There were no significant differences among the three glass ionomers tested. PMID- 11475689 TI - The reliability and validity of the RAPIDD scale: readiness assessment of parents concerning infant dental decay. AB - This study assessed the instrument, Readiness Assessment of Parents concerning Infant Dental Decay (RAPIDD), as a measure of parental readiness to change children's dental behaviors. The 38-item instrument based on four constructs (Openness to Health Information, Valuing Dental Health, Convenience/Difficulty, Child Permissiveness) was administered to a random sample of parents of 6-to-36 month old children on Saipan, Commonwealth of the Northern Mariana Islands, USA. Alpha values for each construct's internal consistency ranged from 0.51 to 0.75. Correlations were found with feeding (e.g., Convenience/Difficulty and receiving bottle (r = .24, p < 0.01)) and oral hygiene (e.g., Valuing Dental Health and tooth-cleaning (r = .27, p < 0.01)). Parental responses were used to categorize respondents into precontemplators (16 percent), contemplators (77 percent), and actions (6.7 percent). This categorization was associated with the number of decalcified and decayed surfaces (alpha 2 = 34.8, p = 0.04; alpha 2 = 65.7, p < 0.01). This suggests that parental readiness is associated with infant dental health and that most parents would be willing to contemplate prevention. PMID- 11475690 TI - Factors influencing whether low-income mothers have a usual source of dental care. AB - Mothers are both the source of dental caries and the managers of children's use of dental care. This is particularly important for the low-income population. The objective of this research was to explore predictors of having a usual source of dental care among low-income women. Eight-hundred-ten mothers (82 percent white, mean age twenty-seven years), drawn from among participants in an ABCD program in one Washington State County completed a study questionnaire. The study examined predisposing, enabling and illness level variables in relation to whether the mother reported a usual source of dental care for herself. Fifty-two percent of the subjects gave positive answers to the dependent variable Do you now have a dentist you can go to if you have a problem? The analysis suggests that mothers that report good oral health and believe in care for their children are five times as likely to have a usual source of care for themselves than mothers who report both poor oral health and more negative attitudes about dental care for children. The results suggest that interventions aimed at child health that ignore the welfare of the mother are likely to be less successful than those that also address the mothers' needs. PMID- 11475691 TI - Bacteria are winning. PMID- 11475692 TI - Behavioral management strategies for young pediatric dental patients with disabilities. AB - Disruptive behaviors, particularly from those lacking in cooperative ability, often are prompted by the need to protest an unpleasant situation and the impulse to protect oneself from perceived danger. Such behaviors, depending on the patient's age and cognitive ability, should be seen as an attempt of the child to cope with a frightening situation. The inherent challenge for both clinician and parent is to avoid unpleasant and unproductive confrontations from the outset, and to create an environment to facilitate the child's ability to accept care, protect the child's self-esteem, foster a positive outlook toward care, and enhance the work quality of dental personnel. In order to use safely any of the modalities for sedation considered [table: see text] in this paper, it is imperative that the clinician receive adequate clinical instruction before private clinical use is attempted. PMID- 11475693 TI - The application of molecular biology. AB - Molecular biology methods have tremendous value not only in the investigation of basic scientific questions, but also in application to a wide variety of problems affecting the overall human condition. Disease prevention and treatment, generation of new protein products, and manipulation of plants and animals for desired phenotypic traits are all applications that are routinely addressed by the application of molecular biology methods. Because of the wide applicability of these methods, they are rapidly becoming a pervasive--some would argue invasive--aspect of our technologically based society. The public concerns that address the application of these methods should be addressed by informed public discussion and debate. While scientists can be extremely critical of the quality, interpretation, and significance of experimental results, they have a rather remarkable tendency to be non-judgmental of the relative social merits of many applications of scientific research. It remains a public responsibility to be sufficiently well-informed to critically assess the merits of applied science research and participate in a communal decision-making process regarding the extent to which a new technology will be allowed to affect society. PMID- 11475694 TI - Purification of nucleic acids by hybridization to affinity tagged PNA probes. AB - The use of affinity tagged PNA capture probes offers an efficient means for the purification of nucleic acids by hybridization. Two different approaches are described. A sequence specific method and a generic method. The sequence specific method requires sequence information on the target and synthesis of a dedicated PNA. It can be used to selectively purify the nucleic acid containing the target from non-related nucleic acids and other cellular components. The generic method uses a "universal" triplex forming PNA and requires no sequence information on the target. It can be used in the bulk purification of large nucleic acids. PMID- 11475695 TI - Peptide nucleic acid (PNA) antisense effects in Escherichia coli. AB - Antisense peptide nucleic acid (PNA) can be used to control cell growth, gene expression and growth phenotypes in the bacteria Escherichia coli. PNAs targeted to the RNA components of the ribosome can inhibit translation and cell growth, and PNAs targeted to mRNA can limit gene expression with gene and sequence specificity. In an E. coli cell extract, efficient inhibition is observed when using PNA concentrations in the nanomolar range, whereas micromolar concentrations are required for inhibition in growing cells. A mutant strain of E. coli that is more permeable to antibiotics also is more susceptible to antisense PNAs than the wild type. This chapter details methods for testing the antisense activities of PNA in E. coli. As an example of the specific antisense inhibition possible, we show the effects of an anti-beta-galactosidase PNA in comparison to control PNAs. With improvements in cell uptake, antisense PNAs may find applications as antimicrobial agents and as tools for microbial functional genomics. PMID- 11475696 TI - PNA biosensors for nucleic acid detection. AB - Biosensor devices, based on the conversion of nucleic acid recognition reactions into useful electrical signals, offer considerable promise for DNA diagnostics. The unique hybridization properties of solution-phase PNA can be extrapolated onto transducer surfaces in connection with the design of remarkably specific DNA biosensors. This article reviews the development of PNA biosensors, and discusses common PNA-biosensing protocols along with their prospects in DNA biosensor technology. PMID- 11475697 TI - Synthesis of infectious viroids and other circular RNAs. AB - Viroids are small autonomously replicating RNAs that share structural features with other subviral circular single-stranded RNAs of plants. Viroids and other circular single-stranded RNAs can be synthesised in vitro by a PCR-based procedure using a simple set of reactions. Two end-to-end primers are selected from a desired region of the viroid, one for the synthesis of the first strand cDNA and another for the production of the second strand DNA. The second primer contains an 18 nucleotide T7 promoter at its 5' end, and is selected such that the G nucleotide at the transcription start site represents a G in the viroid. Linked reverse transcription-PCR results in linear double-stranded DNA consisting of the viroid sequence and the T7 promoter. Run-off transcription of the PCR product allows the synthesis of exact-length linear viroid RNA which can be circularised by T4 RNA ligase following an enzymic modification of the 5' triphosphate to a monophosphate. This procedure results in authentic viroid molecules and obviates the need for construction and cloning of DNA in the form of tandem repeats for infectivity tests. It also allows PCR-based manipulation of circular RNAs, thus greatly simplifying structure-function analyses of viroid molecules. PMID- 11475698 TI - DNA splicing by directed ligation (SDL). AB - Splicing by directed ligation (SDL) is a method of in-phase joining of PCR generated DNA fragments that is based on a pre-designed combination of class IIS restriction endonuclease recognition and cleavage sites. Since these enzymes cleave outside of their recognition sites, the resulting sticky end can have any desired sequence, and the site itself can be removed and does not appear in the final spliced DNA product. SDL is based on the addition of class IIS recognition sites onto primers used to amplify DNA sequences. Cleavage of the PCR products results in elimination of the recognition site-containing flanking sequences and leaves the DNA fragments crowned with protruding ends. With careful design of the sticky ends, several segments can be ligated together in a predetermined order in a single reaction. SDL requires fewer rounds of amplification than overlap extension methods, and is particularly useful for creating a series of recombinants that differ in one segment. PMID- 11475699 TI - The yeast two-hybrid system: criteria for detecting physiologically significant protein-protein interactions. AB - In vivo transcription-based assays for protein-protein interactions such as the two-hybrid system are powerful methods for identifying novel proteins based on their physical association with known proteins of biological interest, or for characterizing the degree and nature of interactions between sets of proteins. Because of the complexity inherent in assays taking place within a living organism, a key issue for the effective use of two-hybrid approaches is the ability to determine whether apparent interactions are likely to be physiologically relevant. In this article, a number of the different two-hybrid systems currently available for use will be reviewed. Then, taking as a model one such system, the Interaction Trap, examples of different reagents for use in varying the affinity range of detectable interactions will be outlined. Also set forth are a number of protocols to establish an appropriate set of conditions for either screening a library or analysing the interaction phenotype between protein sets. Finally, a number of general guidelines are suggested for trouble-shooting two-hybrid results, and for eliminating falsely positive interactions. PMID- 11475700 TI - A PCR-based method for isolation of genomic DNA flanking a known DNA sequence. AB - We describe a simple PCR-based method for the isolation of genomic DNA that lies adjacent to a known DNA sequence. The method is based on the directional cloning of digested genomic DNA into the multiple cloning site of a pUC-based plasmid to generate a limited genomic library. The library is plated onto a number of selective LA plates which are incubated overnight, and recombinant plasmid DNA is then isolated from resistant colonies pooled from each plate. PCR amplification is performed on the pooled recombinant plasmid DNAs using primers specific for the pUC vector and the known genomic sequence. The combination of efficient directional cloning and bacterial transformation gives relative enrichment for the genomic sequence of interest and generates a simple DNA template, enabling easy amplification by PCR. PMID- 11475701 TI - Identification of lactobacilli and bifidobacteria. AB - Selective culture media and phenotypic tests enable lactobacilli to be differentiated from morphologically similar bacteria. The accurate identification of Lactobacillus species can be accomplished by reference to 16S rRNA gene sequences. Species-specific, PCR primers that target the 16S-23S rRNA spacer region are available for a limited number of Lactobacillus species. Molecular methods for the comprehensive identification of Bifidobacterium species are not yet available. Only DNA-DNA reassociation provides a reliable means of species identification for this genus at present. Bifidobacteria can be differentiated from morphologically similar bacteria by the use of genus-specific, PCR primers or oligonucleotide probes. PMID- 11475702 TI - Cell biological studies of the prion protein. AB - Studying PrPC and PrPSc in cell culture systems is advantageous because such systems contain all the organelles, membranes, and molecular cofactors that are likely to play an important role in the biology of the proteins. Using cultured cells expressing PrPC, we have discovered that this isoform constitutively cycles between the cell surface and an endocytic compartment, a process that is mediated by clathrin-coated pits and a putative PrPC receptor. We have also constructed stably transfected lines of CHO cells that express PrP molecules carrying mutations that are associated with familial prion diseases. The mutant PrP molecules in these cells are spontaneously converted to the PrPSc state, a phenomenon which has allowed us to analyze several key features of prion formation. PMID- 11475703 TI - Endonuclease-mediated long PCR and its application to restriction mapping. AB - The polymerase chain reaction (PCR) is the most widely used technique for the study of DNA. Applications for PCR have been extended significantly by the development of "long" PCR, a technique that makes it possible to amplify DNA fragments up to 40 kb in length. This article describes two novel applications of the long PCR technique, one which simplifies restriction mapping and another which enhances amplification specificity and yield. The same primers used to perform the long PCR amplification can be used as probes to perform restriction mapping of the DNA fragment amplified. Restriction digestion performed prior to long PCR amplification can be used to selectively suppress the amplification of members of families of closely related DNA sequences, thereby making it possible to selectively amplify one of a group of highly homologous sequences. These two complimentary techniques, both involving use of the long PCR paired with restriction digestion, have potential application in any laboratory in which PCR is performed. PMID- 11475704 TI - An introduction to peptide nucleic acid. AB - Peptide Nucleic Acid (PNA) is a powerful new biomolecular tool with a wide range of important applications. PNA mimics the behaviour of DNA and binds complementary nucleic acid strands. The unique chemical, physical and biological properties of PNA have been exploited to produce powerful biomolecular tools, antisense and antigene agents, molecular probes and biosensors. PMID- 11475705 TI - Power toothbrushes. Efficacy of a unique gum protection system with an interproximal tip. Introduction. PMID- 11475706 TI - The efficacy of the Philips-Jordan 2-Action plaque remover regarding interproximal access. AB - A laboratory study was performed to assess the interproximal access efficacy of the Philips-Jordan 2-Action plaque remover. This electric toothbrush combines a pressure-control system, and a newly designed brush head with an oscillatory rotary brush and a smaller interproximal brush (the "Active Tip"). The pressure control system is designed to alert the user when too much brushing force is being exerted. The Active Tip brushing head is designed to increase access to the interproximal areas between the teeth. To investigate the added value of the Active Tip, this laboratory investigation measured interproximal access efficacy both with the Active Tip and without it, under identical conditions. The results demonstrated that the interproximal access of the toothbrush was significantly (p < 0.01) enhanced by the presence of the Active Tip. PMID- 11475707 TI - Plaque removal efficacy of two rotating/oscillating electric toothbrushes. AB - This study compared the plaque removal efficacy of the Philips Jordan HP510 and Braun Oral-B D9 electric toothbrushes in two clinical trials. In Trial I, a single-blind randomized split-mouth study, 24 subjects received an oral prophylaxis and abstained from all oral hygiene procedures for 24 hours before each quadrant was professionally brushed for 30 seconds. Plaque was scored before and after brushing using the Turesky-modified Quigley and Hein index (Q&H) and the approximal plaque index (API). Trial II was a two-week, single-blind balanced cross-over study comprising 54 subjects. At baseline, the subjects received an oral prophylaxis and instruction in the use of the assigned brush. Two weeks later, they returned to the clinic with 24-hour plaque accumulation. Plaque (Q&H, API) was scored before and after supervised brushing (30 sec/quadrant). After a prophylaxis, the subjects received the other brush with instructions on use, and the procedure was repeated. In Trial I, plaque was removed almost equally well by both brushes, with the HP510 and the D9 reducing Q&H from 1.46 and 1.47 (baseline) to 0.53 and 0.58, respectively, on average over all teeth. For the premolars and molars in particular, use of the HP510 resulted in statistically significantly lower Q&H scores than the D9 (0.70 vs. 0.81). In Trial II, the HP510 resulted in significantly better plaque scores than the D9 on Q&H (0.55 vs. 0.67) and API (23% vs. 28%) for all teeth. The buccal surfaces contributed the largest difference in Q&H. Statistically significant differences were also found in scores of the posterior teeth for Q&H (0.75 vs. 0.86) and API (31% vs. 38%). In conclusion, both rotating/oscillating electric toothbrushes were highly effective in plaque removal. The HP510 removed more plaque than the D9, particularly from the posterior teeth. It appeared that the smaller height and the Active Tip of HP510's brush head could have resulted in better posterior and approximal access. PMID- 11475708 TI - Short-term clinical study comparing supragingival plaque removal and gingival bleeding reduction of the Philips Jordan HP735 to a manual toothbrush in periodontal patients in a maintenance program. AB - The Philips Jordan HP735 was compared to a manual brush for plaque removal efficacy and reduction of gingival bleeding. Subjects in a periodontal maintenance program were randomly divided into two groups; Group I (n = 27), average age 36.9 years, brushed with the manual brush; and Group II (n = 22), average age 32.9 years, brushed with the Philips Jordan HP735 electric brush. A dichotomous plaque and bleeding index was used at six sites on all teeth at baseline, three, six and nine weeks. The subjects did not use any other cleaning devices during the study. No significant statistical difference in plaque score or bleeding score was found between the two groups at baseline. Plaque scores did not statistically significantly decrease over time in either group, and there was no significant difference in plaque removal between groups during the study. The bleeding index decreased significantly in the electric toothbrushing group; however, due to the large variation in bleeding scores between subjects, the difference in the number of bleeding sites was not statistically significant between the two groups. In conclusion, in a group of periodontal patients in a maintenance phase, using an electric toothbrush did not significantly enhance plaque removal, but did decrease bleeding compared to baseline. The difference in bleeding percentages was not statistically significant compared to a manual brush. PMID- 11475709 TI - Influence of a controlled pressure system on toothbrushing behavior. AB - The use of excessive brushing force has been shown to be a major cause of gingival abrasion. To aid in preventing over-vigorous brushing, the Philips/Jordan electric toothbrush incorporates a Controlled Pressure system (CPS) that causes the brush head to flex back when a toothbrushing force (TBF) in excess of a pre-determined threshold is exerted against the teeth or soft tissues. Two studies (I/II) were conducted to determine whether this mechanical feedback system is sufficiently sensitive to enable users to control their brushing behavior. In Study I, the learning pattern of brushing behavior as a response to the feedback system was evaluated. Seventeen subjects were asked to brush their teeth under observation at least twice a day for a two-week period. During these observations, the number of clicks, as well as the time the brush was pushed "through the click" were recorded. Ten of seventeen volunteers demonstrated a clear learning behavior; the mean number of clicks/minute (for all subjects) was reduced from 10 to 4 after 10 sessions of brushing, and then to 2 or 3 clicks at the end of two weeks. In Study II, 46 subjects used the electric toothbrush with the CPS click force set at various levels between 150-420 g (at 30 g intervals). After a 4-week learning period, the mean TBF was determined in each subject. TBF was most strongly influenced at pre-set click forces between 180 and 270 g. The mean TBF was lowest (about 80 g) when the threshold was set at 210 or 240 g; it then increased (to about 130-140 g TBF) both for smaller and larger values of the threshold setting. Hence, both studies indicate that the Controlled Pressure system is a functional feature that can be used to control the habitual brushing force in a learning period of less than 2 weeks. PMID- 11475710 TI - Plaque removal efficacy of an electric and a manual toothbrush with additional interdental tufts. AB - This clinical study compares the plaque removal efficacy of the Philips/Jordan 2 Action Plaque Remover electric toothbrush with that of the Oral-B 35 Advantage manual toothbrush. In this single-blind randomized split-mouth study, subjects brushed under supervision after having abstained from all oral hygiene for 24 hours. Plaque scored after brushing was used to compare the efficacy of the toothbrushes. Both toothbrushes have special tufts to promote interdental cleaning. It was found that these tufts improve interdental cleaning. Overall, the Philips/Jordan brush removed significantly more plaque than the Oral-B manual brush. PMID- 11475712 TI - [My recognition on integrated traditional Chinese and Western medicine]. PMID- 11475711 TI - A comparative study of two powered toothbrushes and one manual toothbrush in young adults. AB - The aim of this 3-group, 3-treatment, single-blind, parallel group study was to evaluate and compare the efficacies of the Philips/Jordan HP735 powered toothbrush, the Braun/Oral-B D7 powered toothbrush, and the Oral-B Advantage B35 manual toothbrush in young adults (18-25 years). Full mouth mean (SD) modified Turesky Plaque Index (PI) and the Loe & Silness Gingival Index (GI) were recorded at baseline. After 24 hours abstinence from oral hygiene, the PI was recorded and each subject was given one of the test brushes with detailed instructions for use. Each subject then brushed under supervision for 90 seconds, during which time mean (SD) toothbrushing forces (TBF) were recorded. PIs were recorded immediately after brushing, and the subjects were then discharged for 6 weeks to use the allocated toothbrush at home. After 6 weeks, PI, GI and TBF were again recorded. There were no significant differences (ANOVA) for PI and GI between groups at baseline, or for PI following brushing at 24 hours. After 6 weeks, the powered brushes produced lower mean PI (SD) scores than the manual brushes, but the differences were significant only at interproximal sites. Mean GI scores for the HP735 group were similar at baseline and at 6 weeks, although for the other brushes the GI scores actually increased over this period. Mean (SD) TBF (grams/force) at baseline and 6 weeks, respectively, for the brushes were; HP735 233 (205), D7 159 (58), B35 279 (122) (p = 0.026): HP735 194 (86), D7 141 (57), B35 297 (113) (p = 0.0001). The within-group variability for the HP735 TBF reduced considerably over 6 weeks, which is possibly because of the click-force threshold feature of this brush. PMID- 11475713 TI - [Influence of ligustrazine-contained peritoneal dialysate on efficiency of dialysis]. AB - OBJECTIVE: To observe the influence of ligustrazine on efficiency of peritoneal dialysis. METHODS: A prospective, self-controlled study was adopted, 24 stable continuous ambulatory peritoneal dialysis (CAPD) patients were divided into 4 groups according to their peritoneal membrane solute transport types identified by peritoneal equilibration test (PET), the high transport group (H group, 4 cases), the high-average transport group (Ha group, 11 cases), the low-average transport group (La group, 8 cases) and the low transport group (L group, 1 case). After treated with standard dialysis for 2 weeks, patients were treated by peritoneal dialysis with ligustrazine (240 mg/d) contained dialysate for another 2 weeks, and changes on mean net ultrafiltration volume (UF) per exchange, and urea kinetics (KT/V) per week of patients were observed. RESULTS: After ligustrazine treatment, the UF and KT/V raised significantly, P < 0.01. In comparison of groups with different transportation, except change on UF of the H group was insignificant, increment of both parameters before and after treatment were significant (P < 0.01 or 0.05). CONCLUSION: By adding ligustrazine in standard dialysate can enhance the efficiency of peritoneal dialysis, raise the UF, so is worth further study. PMID- 11475714 TI - [Clinical study on effect of Chinese herbal medicine combined with hemodialysis in treating uremia]. AB - OBJECTIVE: To elevate the efficacy of dialysis in uremia patients treated with maintaining hemodialysis. METHODS: Thirty patients of terminal stage uremia were treated with Tongmai oral liquid and rhubarb capsule combined with hemodialysis were observed and compared with a control group of 30 patients treated with hemodialysis alone. The therapeutic course of both groups was 1 month and the observation lasted for 3 months consecutively. RESULTS: The mean time urea nitrogen concentration, protein catabdic rate and the GS index (KT/V value) of the treated group were all better than those of the control group (P < 0.05). CONCLUSION: Uremia patients treated with Tongmai oral liquid and rhubarb capsule combined with hemodialysis were better in full utilization of hemodialysis and life quality of patients in comparison with patients treated with hemodialysis alone. PMID- 11475715 TI - [Clinical study on improving effect of Buyang Huanwu decoction on plasma thromboxane B2, 6-keto-prostaglandin F1 alpha, endothelin and calcitonin gene related peptide in primary nephrotic syndrome patients]. AB - OBJECTIVE: To explore the mechanism of Buyang Huanwu Decoction (BHD) in treating primary nephrotic syndrome (PNS). METHODS: Based on the treatment of prednisone acetate and cytoxan, two groups of PNS patients were treated with aspirin and persantin (western medicine group, 35 patients) and BHD and western medicine (TCM WM group, 35 patients) respectively. The effect on anticoagulation was observed and compared. Plasma levels of thromboxane B2 (TXB2), 6-keto-prostaglandin F1 alpha (6-Keto-PGF1 alpha), endothelin (ET), calcitonin gene related peptide (CGRP) were determined before and after treatment, and at the time of reducing dose and turning to maintenance dose of prednisone. The therapeutic effect of the two groups were also observed. Another group of 30 healthy person was established for control. RESULTS: The difference of TXB2, 6-Keto-PGF1 alpha, ET, CGRP between patients and healthy persons was very significant before treatment (P < 0.001). Above-mentioned 4 parameters improved synchronously with the clinical improvement in the therapeutic course and they were better in the TCM-WM group than those in the western medicine group (P < 0.001) and the complete remission rate of the former group was also higher than that of the latter (62.9% vs 37.1%, chi 2 = 4.63, P < 0.05). CONCLUSION: BHD could improve the therapeutic effect in treating PNS through the mechanism of improving TXB2, 6-Keto-PGF1 alpha, ET and CGRP levels. PMID- 11475716 TI - [Clinical observation on nephrotic syndrome of lupus nephritis treated with integrated traditional Chinese and Western Medicine]. AB - OBJECTIVE: To observe the clinical effect of integrated traditional Chinese and western medicine (TCM-WM) in treating nephrotic syndrome of lupus nephritis (NSLN). METHODS: One hundred and fifty-five patients of NSLN were randomly divided into the TCM-WM group and the control group (treated with western medicine alone). Changes of activity index (AI) of lupus nephritis, urinary protein (UP), plasma albumin (PA), blood lipid (BL) of the patients before and after treatment were observed and compared. And the difference on recurrence rate and adverse reaction rate between the two groups were assessed. RESULTS: The total effective rate of the TCM-WM group was 95%, significantly higher than that of the control group (79%, P < 0.05). The effect in improving AI, UP, PA and BL of the TCM-WM group was superior to that of the control group. The recurrence rate and adverse reaction rate of the former group were 8% and 34% respectively, which was much lower than those of the latter (38% and 68%, P < 0.05). CONCLUSION: TCM-WM treatment is obviously superior to treatment with western medicine alone. PMID- 11475717 TI - [Clinical study of weishu capsule in treating precancerous lesions of chronic atrophic gastritis]. AB - OBJECTIVE: To explore the therapeutic effect and mechanism of Weishu capsule (WSC) in treating precancerous lesions of chronic atrophic gastritis (CAG). METHODS: Seventy-five cases of precancerous lesions of CAG were distributed to two groups randomly, the treated group (45 patients) treated with WSC by taken orally and the control group (30 patients) treated with Weining granule (WNG) orally. The treatment course of both groups were 6 months. The clinical and pathological effects and the changes in scores of atrophy, intestinal metaplasia and its subgroups, dysplasia, carcinoembryonic antigen (CEA) and proliferating cell nuclear antigen (PCNA) expression were observed before and after treatment. RESULTS: The total effective rate was 91% and the effective rate of atrophy, intestinal metaplasia, dysplasia in the treated group were 64%, 67% and 65% respectively, and the respective scores were lowered significantly after treatment. Compared with the control group the difference between the two groups was statistically significant (P < 0.01, P < 0.05). Immunohistochemical test showed that WSC could inhibit the subgroups of intestinal metaplasia, CEA and PCNA expression significantly (P < 0.01). CONCLUSION: WSC could significantly improve the clinical and pathological changes in the precancerous lesions of CAG. The mechanism might be related to the inducing and promoting effect of WSC on the differentiation and maturity of intestinal metaplasia cells and dysplasia cells. WSC could also inhibit and correct the abnormal proliferation of cells. PMID- 11475718 TI - [Determination of gastric emptying time of functional dyspepsia and clinical study on therapeutic effect of Weihuigui decoction on functional dyspepsia]. AB - OBJECTIVE: To observe the gastric emptying time (GET) of and the therapeutic effect of Weihuigui Decoction (WHGD) upon functional dyspepsia (FD). METHODS: GET of 64 FD patients and 20 healthy volunteers was measured by real time ultrasonography. The measurements were performed at fasting, 0, 10, 20, 30, 40 and 50 min after drinking water, the area of gastric antrum, internal diameter of the section of anterior posterior walls in corpus ventriculi fundus junction region, intitating time of gastric contraction and contractive times in 2 min were recorded. Twenty patients selected randomly were treated with WHGD 100 ml, three times a day, for 2 weeks consecutively, to observe the improvement of clinical effect and gastric emptying. RESULTS: GET was delayed in 66% of 64 FD patients as compared with that of control, and it was related with gastric contraction initiating time and contraction times in 2 min. WHGD could improve the clinical symptoms and GET of FD patients. CONCLUSION: WHGD has obvious therapeutic effect in treating FD. PMID- 11475719 TI - [Comparative study on effect of Panax notoginseng and ticlid in treating early diabetic nephropathy]. AB - OBJECTIVE: To explore the ameliorative effect and mechanism of Panax notoginseng (PNG) and ticlid in treating early diabetic nephropathy (DN). METHODS: Fifty eight patients were divided randomly into two groups, 28 patients of the ticlid group treated with ticlid 250 mg orally, once a day and 30 patients of the PNG group treated with PNG 8 ml in 250 ml of normal saline intravenous drip once a day. The therapeutic effect and relative indexes of the two groups were observed and compared. RESULTS: After treatment, in both groups, the thromboxane B2 markedly reduced and was more prominent in the ticlid group (P < 0.05), while the 6-keto-prostaglandin F1 alpha increased obviously, so as to cause a significant lowering of T/K ratio, P < 0.01. Levels of urinary albumin, beta 2 microglobulin and blood alpha 1 microglobulin of both groups were lowered significantly, P < 0.01. A significant positive linear correlation was found in the ticlid group between urinary albumin and T/K ratio (r = 0.41, P < 0.01), as well as in blood alpha 1 microglobulin with T/K ratio (r = 0.34, P < 0.05), while it was not found in the PNG group. CONCLUSION: Ticlid and PNG were beneficial to resume the balance of T/K and improve microcirculation, reduce whole blood viscosity and decrease urinary albumin so as to retard the progress of DN. PMID- 11475720 TI - [Experimental study on effect of Buyang Huanwu decoction on arterial blood platelet activating factor content pre- and post-arterial thrombosis of rats]. AB - OBJECTIVE: To explore the effect of Buyang Huanwu Decoction (BHD) on arterial blood platelet factor (PAF) content in pre- and post-arterial thrombosis of rats. METHODS: Male Wistar rats were randomized into 3 groups, the BHD group treated with BHD 32 g/kg, or with BHD 16 g/kg daily for 14 days consecutively, the control group treated with dexamethasone 0.5-1 mg/kg and the blank group treated with distilled water 10 ml/kg. Arterial thrombosis was duplicated in the animals 2 hours after final medication, the blood content of PAF (by bioassay), dry weight (W) and occlusion time (OT) of thrombus, and weight of thrombus/body weight (TW/BW) ratio were observed. RESULTS: Consecutive BHD treatment could markedly lower the arterial blood content of PAF after thrombosis, increase the OT value of thrombus, reduce the dry weight of thrombus and the TW/BW ratio (P < 0.05), while no influence was found in rats treated with BHD once. CONCLUSIONS: BHD has certain antagonistic effect on pathologic PAF metabolism, and inhibitory effect on pathogenesis and development of arterial thrombosis. PMID- 11475721 TI - [Dynamic effect of tifukang on brain proopiomelanocortin gene expression of exercise-induced fatigue rat]. AB - OBJECTIVE: To study the mechanism of exercise-induced fatigue (EIF) and effect of Tifukang (TFK) from the gene transcription level. METHODS: In situ hybridization was adopted, using biotin-labelled proopiomelanocortin (POMC) cRNA probe and integrating with image pattern analysis, to observe the effect of TFK on changes of POMC mRNA expression in different areas of EIF rat's brain. RESULTS: TFK could significantly strengthen POMC mRNA expression in hippocampal CA1 and hypothalamus and inhibit obviously that in frontal cortex of chronic fatigue rats. In rats managed with long term (7 weeks in total) and high intensity exercise with the speed increased from 15 m/min gradually to 35 m/min, 20 to 25 min/d, after the final exercise, the POMC mRNA expression in frontal cortex and hippocampal CA1 decreased (the latter recovered at 30 minutes after ending exercise but the former was not recovered yet 3 hours after the ending of exercise), while that in hypthalamus increased. TFK could remarkably increase POMC gene expression in the above-mentioned three areas after the final exercise especially prominent at the time of 3 hours after the ending of exercise. CONCLUSION: The dynamic changes of POMC gene expression in different areas of EIF rat's brain were not the same, and TFK displayed an important dynamic regulating effect on it. PMID- 11475722 TI - [Experimental study on inhibitory effect of ganlu xiaodu Dan on coxackie virus in vitro]. AB - OBJECTIVE: To observe the Coxackie virus inhibitory effect of Ganlu Xiaodu Dan (GLXDD), including complete recipe, incomplete recipe and supplemented recipe of it. METHODS: Using tissue culture technique, the maximal non-toxic concentration of drug on cultured cells, the influence of drug on proliferation of virus in cultured cells and the virus inhibitory effect in different time of action were observed. RESULTS: The maximal non-toxic concentration of GLXDD was 1:40 (25 g/L), no matter whether the recipe was complete, incomplete or supplementary. CLXDD in that concentration could inhibit significantly the proliferation of Coxackie virus B2, B3 and B4 strains, the inhibition index being over 2 and the output of these strains treated with complete recipe of GLXDD was lower significantly than that of the control group (P < 0.05). CONCLUSION: GLXDD, including its complete recipe, incomplete recipe and supplementary recipe, could inhibit the replication of Coxackie virus in cultured cells. PMID- 11475723 TI - [Progress in the research on chemical composition and pharmacology of radis Achyranthes bidentata]. PMID- 11475724 TI - [Clinical and experimental study on the Houpu Mahuang oral liquid in treating bronchial asthma]. AB - OBJECTIVE: To probe into the curative effect and mechanism of action of the Houpu Mahuang oral liquid (HPMH) in treating bronchial asthma. METHODS: The pulmonary function and level of serum IgE of the patients were determined before and after HPMH treatment. The anti-allergic reaction and relieving spasm effect of HPMH in animal were also observed. RESULTS: HPMH is not only effective in improving clinical symptoms and pulmonary function (P < 0.05), so as to lower the abnormal raising of IgE (P < 0.01), but also has a notable inhibitory effect for passive allergic reaction of rat's skin, it can relieve spasm. CONCLUSIONS: HPMH is effective in relieving spasm, antiallergic reaction, it can improve the symptoms of bronchial asthma. PMID- 11475725 TI - [Therapeutic effect of Shengmai injection on respiratory function in chronic obstructive pulmonary disease]. AB - OBJECTIVE: To explore the therapeutic effect of Shengmai injection (SMI) on respiratory function in chronic obstructive pulmonary disease (COPD). METHODS: Twenty patients with COPD were continuously treated with SMI by intravenous drip in 14 days and examined the lung vital capacity (VC), forced vital capacity (FVC), forced vital capacity of the first second (FEV1), FEV1/FVC, maximum voluntary ventilation (MVV), maximum inspiratory pressure (MIP), load respiratory time (LT), 6 minute walk distance (6 MWD), arterial blood gas analysis, and Brog dyspnea scale before and after treatment respectively and compared with itself or control group. RESULTS: After treatment all parameters of the treatment group were better than those of control group or itself before treatment except for FVC and FEV1/FVC. There were significant difference (P < 0.05). CONCLUSION: SMI can improve respiratory function in COPD. So it is an effective rehabilitative measure in treating COPD. PMID- 11475726 TI - [Clinical and experimental study on Fuzheng Baozhen Decoction enhancing effect of radio- and chemotherapy for malignant tumors]. AB - OBJECTIVE: To study on Fuzheng Baozhen Decoction (FZBZD) enhancing effect of radio- and chemotherapy for malignant tumors. METHODS: One hundred and Seventeen cases of malignant tumors treated with chemotherapy or/and radiotherapy, 55 cases treated together with FZBZD (group A), comparing with 30 cases treated with radio and chemotherapy plus Zhenqi Fuzheng Granules (group B) and 32 cases radio- and chemotherapy alone (group C). Mechanism of FZBZD enhancing effect of chemotherapy on transplanted human lung adenocarcinoma (SPC-A-1) and sarcoma (S180) bearing mice was conducted. RESULTS: Effective rates (CR + PR) of group A, B, C were 63.6%, 43.3%, 37.5% respectively, that of group A was the best (P < 0.05). The survival quality of life was improved best in group A (P < 0.05). After being treated, the level of the peripheral blood (WBC, Hb, PLT) of group A was the highest (P < 0.05); CD3, CD4, NK activity, interleukin-2 were also improved significantly in group A (P < 0.01). Animal experiment showed that FZBZD could improve chemotherapy effect of inhibitory action on tumor growth (P < 0.01), increase cAMP/cGMP ratio (P < 0.01) by adding cAMP level in cancer tissue, and enhance of G0/G1 phase cells and decrease S phase cells. CONCLUSIONS: FZBZD inhibited tumor growth and enhanced the effect of radio- and chemotherapy by improving immune and hematopoietic function, cAMP/cGMP ratio and stagnating tumor cells in G0/G1 phase. PMID- 11475727 TI - [Diagnostic significance of topical image of infrared thermograph on the patient with lumbar intervertebral disc herniation--a comparative study on 45 patients and 65 normal control]. AB - OBJECTIVE: To analyse and explore the diagnostic significance of infrared thermography on the patients with lumbar intervertebral disc protrusion. METHODS: Forty-five hospitalized cases under conservative treatment (mainly manipulation) and 65 controls were sequentially selected. Both groups were examined with infrared thermograph on lower back and lower extremities as well as physical examination before and after treatment. The study statistically analysed the temperature difference between the involved and healthy parts at different areas; and also compared the change of the temperature difference after the treatment to the decreasing scores of physical examination afterward. RESULTS: The statistic result showed that the temperature difference between two sides on patients group was significantly higher than those of control group. The further analysis shows that the temperature difference at posterior femur area in the patient's group correlated significantly to the severity of clinical signs caused by nerve root irritation. CONCLUSIONS: The thermograph could only be considered as a reference in the diagnosis of lumbar disc protrusion. It is helpful to discover the severity of radical signs according to the posterior femoral region of hypo- (or hyper) thermogram. PMID- 11475728 TI - [Clinical study of tubal pregnancy treated with integrated traditional Chinese and Western medicine]. AB - OBJECTIVE: To explore the therapeutic effect and the mechanism of tubal pregnancy (TP) treated with integrated traditional Chinese and Western medicine (TCM-WM). METHODS: Seventy-eight patients suffering from TP were divided into two groups at random, 50 cases of treated group treated with the method of integrated Methotrexate (MTX) and Ectopic Pregnancy 2 (EP2) decoction, and the other 28 cases treated with MTX alone for control. RESULTS: The nonsurgical method cure rate of the treated group (98.0%) was higher than that of the control group (75.0%, P < 0.01). The intrauterine pregnancy rate of the treated group was 80.0%, that of the control group was 42.9% (P < 0.01). The recurrent TP rate of the treated group (4.0%) was lower than that of the control group (14.2%, P < 0.01). The serum beta-HCG normalizing time of the treated group (25.0 +/- 7.8 days) was shorter than that of the control group (31.9 +/- 6.7 days, P < 0.05). The total disappearance time of TP mass was 1.2 +/- 0.7 months in the treated group, and in the control group was 5.6 +/- 3.7 months (P < 0.01). After treated by EP2 decoction for 4-8 weeks, the deformability of erythrocyte was increased, the aggregation of erythrocyte, the whole blood viscosity and the blood plasma viscosity were reduced. CONCLUSIONS: The integrated method can effectively increase nonsurgical cure rate, tubal patency rate and intrauterine pregnancy rate, decrease recurrent TP rate in treating TP. PMID- 11475729 TI - [Study on treatment of blood stasis syndrome of chronic prostatitis using prostatitis decoction and its extract capsule]. AB - OBJECTIVE: To test and verify the effects of prostatitis decoction and its capsule on the treatment of chronic prostatitis with blood stasis Syndrome, and its therapeutic mechanism. METHODS: Five hundred and sixty-one Patients were treated for 2 months, decoction group 322 cases, and capsule group 239 cases. As control group, 95 cases were treated with Qianliekang tablets. Expressed prostatic secretion(EPS)-pH and Zn content were measured before and after treatment. Observation on hemorheology and microcirculation disturbance with animal experiments were conducted. RESULTS: The cure rate of Prostatitis decoction and capsule groups was 65.8%, and 54.4% respectively, that of control group was only 17.9%. EPS-pH and Zn were markedly improved after Prostatitis decoction treatment. Experimental work revealed that Prostatitis decoction and its capsule had lowered the whole blood viscosity, thrombocyte adhesiveness rate and dry weight of thrombus in blood stasis rabbit model; and on rats model of microcirculation disturbance caused by adrenalin, the Prostatitis decoction and its capsules had obviously delayed the occurrence of reduction and stopping of blood flow in arterioles. CONCLUSION: Prostatitis decoction (capsule) was effective in treating chronic prostatitis. PMID- 11475730 TI - [Relationship between Syndrome-Type in TCM and four trace urinary proteins in patients with bronchial asthma]. AB - OBJECTIVE: To explore the possible relationship between Syndrome-Type in TCM and the 4 trace urinary proteins in patients with bronchial asthma. METHODS: The 4 kinds of trace urinary proteins in 217 patients with asthma were measured by radioimmunoassay, and that in 66 normal subjects were compared. RESULTS: The results showed that there were increments of urinary albumin, urinary IgG, urinary beta 2-microglobulin and urinary Tamm-Horsfall protein in patients with asthma; and the gradual increase tended to be accompanied by the continuous aggravation of patient's condition on TCM Syndrome-Type. Furthermore, these datas proved that the 4 trace urinary proteins might be the most sensitive indicators for detecting impaired renal functions than serum blood urea nitrogen and creatinine. CONCLUSIONS: There were the objective material foundations on TCM Syndrome-Type in asthmatics, which consisted of renal glomerular dysfunctions to filter out albumin and IgG, renal tubular dysfunction to reabsorb beta 2 microglobulin, and to synthesize and excrete Tamm-Horsfall protein. PMID- 11475731 TI - [Clinical and experimental study on using Cassia angustifolia extract as enema after abdominal operation]. AB - OBJECTIVE: To investigate the curative effect and mechanism of using Cassia angustifolia extract (CAE) in treating gastrointestinal tract dysfunction after abdominal operations. METHODS: Enema administration of CAE (Clyster method) was used. RESULTS: The result of 130 patients was very effective in reducing the rate of gastrointestinal decompression, accelerating the restitution of borborygmi and the time of exhaustion. Animal experiment showed the CAE function is very obvious in enhancing the bowel movement of rats (P < 0.05). It can enhance peristalsis and contraction amplitude of vibration in the isolated ileum of rats (P < 0.05). It can push on the charcoal powder in intestinal tract of mice obviously (P < 0.05). CONCLUSION: CAE could regulate disordered function of gastrointestinal tract after abdominal operations. PMID- 11475732 TI - [Effect of Salvia Miltiorrhiza injection on umbilical artery hemodynamics during fetal distress in ewe]. AB - OBJECTIVE: To study the effect of Salvia Miltiorrhiza injection (SMI) on umbilical artery hemodynamics during fetal distress in ewe. METHODS: Intrauterine surgery were performed at 116-125 gestational days in 3 pregnancy sheep for insertion of vascular catheters at abdominal aorta from femoral artery and for implantation of a electromagnetic flowmetre probe around umbilical artery. Gelatin microsphere injection to abdominal aorta through vascular catheter for obstruction of placental-fetal circulation. 2 ml of SMI was used 6 times(2 times each sheep), and the signs of umbilical artery blood flow and wave forms of fetal abdominal aorta blood pressure were recorded before and after injections of gelatin microsphere or SMI. The fast Fourier translation (FFT) was used to turn signs in time domain into powers spectral of input impedance in frequency domain for resistance and characteristic impedance of umbilical artery. RESULTS: After the SMI was injected into fetal abdominal aorta, the resistance of umbilical artery was significantly reduced (P < 0.05), and the changes of resistance was significantly relative with the resistance before SMI injected (r = -0.85, P < 0.001). But there were no significantly different impedance between before and after SMI treatment (P > 0.05). CONCLUSIONS: The SMI to fetal abdominal aorta can decrease umbilical artery blood flow resistance and the changes of resistance was significantly relative to that before SMI treatment. PMID- 11475733 TI - [Effects of subcutaneous injection with different doses of ginseng on the rat model of chronic Pseudomonas aeruginosa pneumonia]. AB - OBJECTIVE: To observe the effect of ginseng extracts on a rat model of chronic Pseudomonas aeruginosa (PA) pneumonia. METHODS: Subcutaneous injection with four different doses of ginseng aqueous extracts to the model rats for two weeks. RESULTS: Milder macroscopic lung pathology and lower incidence of lung abscesses were found in all the four groups received different doses of ginseng treatment compared to the control group (P < 0.05). Histopathology of the lungs was negatively correlated with the ginseng doses (r = -0.95, P < 0.01). The mast cell number in the lung foci in group I, II and IV was significantly lower than that in the control group (P < 0.05). Bacterial clearance in group II and group III was enhanced significantly (P < 0.05) compared with that in the control group. The specific antibody responses were down-regulated in group II, III and IV (P < 0.01), and IgM level in group I was lower compared to the control group (P < 0.01). Serum IgM level was positively correlated with the ginseng doses (r = 0.93, P < 0.05). IgG2a titer in group I was higher than that in the control group (P < 0.05). Serum IgG2a level was positively correlated with the ginseng doses (r = 0.90, P < 0.02). Two weeks after PA lung infection, the serum level of IgG against PA, aeruginosa alginate in group I, II, and III was higher than that in the control and group IV (P < 0.05), and the anti-alginate IgA level in group I, III and IV was lower than that in the control (P < 0.05). CONCLUSIONS: The ginseng dose of 2.5 mg/kg was the best dosage in the present study, and it showed an effect of stimulating T helper type I response. PMID- 11475734 TI - [Effect of different prescriptions for tonifying kidney on RNA transcription activity in isolated nuclei and protein synthesis activity in free-cell extracts of the different organs of presenile mice]. AB - OBJECTIVE: To study the anti-aging mechanism of Zuoguiyin (a prescription for tonifying Kindney Yin) and Yishenbao (a prescription for tonifying Kidney Yang) in presenile mice. METHODS: Using male, 14 months old presenile mice as a model of Kidney-Asthenia, the effects of Zuoguiyin and Yishenhao on RNA transcription activity in isolated nuclei and protein synthesis activity in free-cell extracts of the liver, brain, kidney and testes of the presenile mice were observed. RESULTS: RNA transcription activity in isolated nuclei and protein synthesis activity in free-cell extracts of the four organs of the presenile mice group were distinctly lower than those of young mice group (3 months old). After above mentioned two prescriptions were given to the presenile mice for two months, RNA transcription activity in isolated nuclei and protein synthesis activity in free cell extracts of the four organs of the two treated groups were obviously higher than those of control group (P < 0.05 or P < 0.01), but there was no difference between two treated groups. CONCLUSIONS: Both Zuoguiyin and Yishenbao could promote RNA transcription activity in isolated nuclei and protein synthesis activity in free-cell extracts of Kidney-Asthenia and presenile mice. PMID- 11475735 TI - [Effect of Shenmai injection on morphological structure of kidney and peritoneum in rats with 5/6 nephrectomy]. AB - OBJECTIVE: To study the protective effect of Shenmai injection (SMI) on kidney and peritoneum in rats with 5/6 nephrectomy. METHODS: By using cell morphological quantification, light microscopy and scanning electron microscopy, the morphological structure changes of peritoneum and kidney in rats with 5/6 nephrectomy caused chronic renal failure model after the intramuscular or intraperitoneal injection of SMI were observed. RESULTS: The SMI injection could relieve glomerular sclerosis, promote the recovery and proliferation of injured peritoneal mesothelial cell. The effect of the SMI intraperitoneal injection is better than that intramuscular of injection. CONCLUSION: The SMI has the protective effect on kidney and peritoneum in rats with 5/6 nephrectomy. PMID- 11475736 TI - [A glance on dynamic changes of health care in USA]. PMID- 11475737 TI - [Exploration on music therapy]. PMID- 11475738 TI - [T-lymphocyte immune status of children with recurrent respiratory tract infection and progress in its treatment with traditional Chinese and western medicine]. PMID- 11475739 TI - [The comparative study on approach of regulating homeostasis in human body with traditional Chinese medicine. On one of the integration points between Chinese and Western medicine]. PMID- 11475740 TI - [Treatment of hemorrhoid of III, IV stage by four steps injection of xiaozhiling]. AB - OBJECTIVE: To find a new method of non-operation treatment of hemorrhoid in III, IV stage. METHODS: Based on the principle "sore herbs can astringent and the puckery can control prolapse" and precious experience in treating hemorrhoid, four step injections of Xiaozhiling (XZL, preparation made from gallnut and alum chiefly) were injected into the branches of rectal superior artery, the submucosa, the mucous lamina propria and the sinusoid veins, to cause sclerosis and atrophy of hemorrhoid by obliterating branches of rectal superior artery. RESULTS: 21,361 cases were treated from 1987 to 1996. Among them 21,148 (99.00%) were cured, 203 (0.95%) improved and 10 (0.05%) ineffective and for whom surgical operation was used. Of the 620 cases who were followed up for 3 years, only 6 cases (0.97%) relapsed. CONCLUSION: This method is a relative ideal non-operative approach in the treatment of hemorrhoid of stage III and IV. PMID- 11475741 TI - [Clinical observation on artificial shexiang baoxin pill in treating 112 patients of coronary heart disease with angina pectoris]. AB - OBJECTIVE: To test and verify the effect of artificial Shexiang Baoxin Pill (SXBXP, a heart protecting musk pill). METHODS: Two hundred and twenty-five patients of coronary heart disease (CHD) with angina pectoris were randomly and blindly assigned to the study group (112 cases) and the control group (113 cases). The two groups were well matched in age, sex, angina type, complication and grade of cardiac function. Two pills of artificial or natural SXBXP were given to each cases of the two groups for 3 times every day orally, for 2 weeks, respectively. RESULTS: Both the artificial and natural SXBXP could not only reduce significantly the frequency of angina episode and the daily consumption of nitroglycerin, they also could result in remarkable improvement of NST, sigma ST and exercise tolerance, as well as significant reduction of rate-pressure product. The total symptomatic effective rate of the two groups was 86.6% and 89.4% respectively, and the ECG effective rate, 52.7% and 57.5% respectively (P > 0.05), and the incidence of headache of the two groups was one for eoch. In addition, the onset of action of both agents was rapid, 88.5% of the study group and 78.6% of the control group were within five minutes. The sustained time of action of the two groups was also similar, i.e. 4.9 +/- 4.4 h vs 5.4 +/- 3.1 h. CONCLUSION: The clinical efficacy and side effect of artificial SXBXP were very similar to those of natural SXBXP. PMID- 11475742 TI - [Effect of garlicin on unstable angina pectoris and its relationship with blood lipid and GMP-140]. AB - OBJECTIVE: To observe the clinical effect of Garlicin on unstable angina pectoris (UAP) and to explore the relationship between Garlicin's effect and Syndrome Differentiation of TCM. METHODS: Fifty-five patients of UAP were divided randomly into the Garllcin group (34 cases) and the control group (21 cases) and classified based on Syndrome Differentiation of TCM into Cold Syndrome type, Heat Syndrome type, severe blood stasis (SBS) type or mild blood stasis (MBS) type. Garlicin 60 mg or nitroglycerin 5 mg were given to the two groups respectively by intravenous drip for 10 days as one therapeutic course. The curatic effect was evaluated by changes of symptoms and electrocardiogram, and some indexes, such as blood lipid, lipoprotein, apolipoprotein and GMP-140 among different groups and types were compared. RESULTS: The difference of effective rate between the Garlicin group and the control group was insignificant. In the Garlicin group, the effective rate of Cold Syndrome type was higher than that of Heat Syndrome type (P < 0.01), and HDL/LDL ratio and apoA1 level raised markedly in the former type (P < 0.05) while an oppositive trend revealed in the Heat Syndrome type. Garlicin showed a higher effect on SBS type than that on MBS type and cause GMP 140 of the MBS type decreased obviously. CONCLUSION: Garlicin is effective on UAP patients and its effect was more evident on those with Cold Syndrome type or SBS type. Its mechanism may be related to its effect in improving blood lipid and enhancing activation of platelet. PMID- 11475743 TI - [Observation on effect of shenmai injection in treating patients of congestive heart failure]. AB - OBJECTIVE: To observe the clinical therapeutic effect of Shenmai Injection (SMI) in treating patients of congestive heart failure. METHODS: Effect of 16 cases treated with SMI or potassium magnesium aspartate was observed by randomized crossover method and compared. RESULTS: By using SMI for 2 weeks, patients' left ventricular ejective fraction (LVEF) could be increased from 29.5 +/- 9.0 to 36.6 +/- 10.2 (P < 0.05), the heart function of 68.75% patients was improved, no side effects or toxicity was observed. Potassium magnesium asparate could improve heart function in 37.50% patients only but with no effect on LVEF. CONCLUSION: SMI is an effective and safe therapy in treating patients with congestive heart failure. PMID- 11475744 TI - [Clinical study on chanlibao in accelerating second stage of labor]. AB - OBJECTIVE: To find out a new way to strengthen the uterine contraction and shorten the second stage of labor, for improving the prognosis of mother and newborn. METHODS: Primiparae observed were divided into three groups randomly and Chanlibao (CLB, a preparation of Chinese herbal medicine) or oxytocin (OTC) was given to the CLB group (80 cases) and the OTC group (52 cases) respectively, to the third group, as a control group (29 cases) of natural labor, without any drug was given. Time of second stage of labor (TSL) and prognosis of mother and newborn of different groups were observed and compared. And intrauterine pressure (IP) and fetal heart rate were monitored by means of electronic-monitor. RESULTS: TSL and postpartum hemorrhage in the CLB group were less than those in the control group. The average IP in former was higher than that in latter, but with no difference as compared with the OTC group. No side effect of CLB was found. CONCLUSION: CLB could obviously strengthen uterine contraction and shorten the TSL, it is cheap, convenient and with no side-effect, so it is a new safe, effective method for improving prognosis of primiparae and newborn, especially for those nonindicated in using oxytocine or infusion of fluid profusely. PMID- 11475745 TI - [Clinical observation on effect of yangjing decoction in promoting follicular growth]. AB - OBJECTIVE: To investigate the effect of Yangjing Decoction (YJD, a Chinese prescription of replenishing Kidney essence) in promoting follicular growth and its influence on blood supply of ovary and uterus. METHODS: Twenty-two cases with secondary amenorrhea and 4 cases of infertility with infrequent menstruation caused by follicle maldevelopment were treated with YJD. The blood supply of ovary and uterus in 7 cases was measured by colour Doppler before and after treatment. RESULTS: Of the 116 therapeutic cycles in 26 cases, there were 69 menstrual cycles (59.48%), 37 follicular maturation cycles (31.90%), 21 ovulation cycles (18.10%), and the follicular maturation rate was 84.62%, The disturbance in blood supply of ovary and uterus was improved remarkably along with follicular development by YJD in 7 cases. CONCLUSION: YJD could improve follicular development and blood supply of ovary and uterus. PMID- 11475746 TI - [Study on mechanism of bushen yugong granule in treating uterine dysgenesia]. AB - OBJECTIVE: To explore the effective mechanism of Bushen Yugong Granule (BSYGG, a Chinese medicine preparation) in treating infertility due to uterine dysgeneia. METHODS: (1) Clinical study: Serum level of sex hormone and endometrial content of estradial receptor (ER) and progesterone receptor (PR) in follicle period of 60 patients (the tested group) were measured and compared before and after treatment, and the data were also compared with those of 70 normal women (the control group). (2) Experimental study: Rats were divided into the control group and 3 tested groups fed with different dose of BSYGG. Pathologic examination of pituitary, ovary and uterus of rats after BSYGG treatment, and serum levels of ER, PR were measured. Also, an estrogen activity experiment was adopted in some young cophorectomized rats. RESULTS: (1) Clinical study, the difference of serum sex hormone levels between the tested and the control group was insignificant. The percentage of patients in tested group with endometrial ER content over ++ was lower than that in the control group (46.9% vs 72.4%) significantly (P < 0.05), after treatment, it arised to 65.4% in test group, the change after treatment was significant (P < 0.05). (2) Experimental study: The number of follicle, weight of uterus and serum E2 level increased significantly after BSYGG treatment. The endometrial content of ER in the control group was 29.25 +/- 17.03%, while in the tested group, it was 67.75 +/- 24.56%-83.00 +/- 9.87% after treatment, the difference between the two groups was significant (P < 0.005). CONCLUSION: BSYGG has an estrogen-like activity, it could increase the endometrial content of ER, enhance the effect of estrogen on uterus and promote the uterine development, so as to elevate the fertilization rate. PMID- 11475747 TI - [Study on relationship between ovulation inducing effect of drug-acupuncture and endometrial contents of estradiol receptor and progesterone receptor]. AB - OBJECTIVE: To study the effect of Chinese herbal medicine for replenishing Kidney combined with acupuncture in treating anovulation and infertility, and the relationship between its ovulation inducing effect and endometrial contents of estradiol receptor (ER) and progesterone receptor (PR). METHODS: Twenty-nine cases were treated with replenishing Kidney drugs combined with acupuncture for 1 3 months. Patients' ER and PR were measured by immunohistochemical method. And patients were classified according to PR content into PR positive group and mild PR positive group. RESULTS: Fifteen cases of PR positive group, completed treatment for 45 cycles, among them, 40 cycles showed ovulation, the ovulation rate being 88.89%. Ten in 14 cases, who complicated with infertility, became pregnant, the pregnant rate being 71.43%. While in 11 cases of PR mild positive group, 9 complicated with infertility, completed treatment for 33 cycle, 10 cycles showed ovulation (30.30%), and pregnant rate 22.22% (2/9). The difference between the two groups was significant (P < 0.01). CONCLUSION: The replenishing Kidney drugs combined with acupuncture treatment could result a good effect in treating infertility due to anovulation, especially on those with high endometrial PR content. PMID- 11475748 TI - [Preliminary study on therapeutic effect of oxymatrine in treating patients with chronic hepatitis C]. AB - OBJECTIVE: To evaluate the efficacy of oxymatrine in treating chronic hepatitis C and its mechanism. METHODS: Forty-three patient were divided randomly into the treated group (20 cases) and the control group (23 cases). The treated group was given oxymatrine 600 mg per day intramuscularly, and the control group was given the general liver protective agents such as vitamins. The therapeutic course of both groups was 3 months. RESULTS: HCVRNA of 8 in 17 cases (47.1%) of the treated group converted to negative, while in 18 cases of the control group, the negative conversion only took place in 1 patient (5.6%), the negative conversion rate was significantly higher in the treated group than that in the control group (P < 0.05). The normalization rates of serum alanine transaminase (ALT) of the treated group after 1 month and 2 months treatment was higher than that of the control group, but after 3 months treatment, the normalization rates of the two groups were not different significantly. Plasma level of soluble interleukin-2 receptor and serum level of collagen type IV in the treated group were lowered significantly after treatment, but in the control group, there were no significant change, the difference between the two groups was significant (P < 0.01, P < 0.05). CONCLUSION: Oxymatrine is effective in inhibiting proliferation of HCV, antagonisting liver fibrosis and regulating immune reaction of the host, so it could be a safe, effective drug in treating chronic hepatitis C. PMID- 11475749 TI - [Origin of nitric oxide synthase positive nerve fibers at zusanli area in rats]. AB - OBJECTIVE: To find the origin of nitric oxide synthase (NOS) positive nerve fiber at the area around Zusanli point(St. 36). METHODS: The combined method of retrograde transport of horse radish peroxidase and nicotinamide adenosine dinucleotide phosphate diaphorase were adopted. RESULTS: Some peripheral processes of NOS positive neurons were distributed in the Zusanli point from the ganglia of L4 to S1, and some were projected from the lamina IX of L4 to S1 in spinal cord. CONCLUSION: The distributions of NOS positive nerve fibers in the, Zusanli area might be one of the morphological foundations of acupuncture effect of Zusanli point. PMID- 11475750 TI - [Quantitative study on synergistic effect of radix astragali A6 and acyclovir against herpes simplex virus type I by polymerase chain reaction]. AB - OBJECTIVE: To evaluate the synergistic effect of Radix Astragali A6(A6) and acyclovir (ACV) in antagonizing herpes simplex virus type I (HSV1). METHODS: The synergistic effect was measured by competitive polymerase chain reaction (PCR) and compared with that measured by traditional cytopathic effect (CPE) inhibition method. RESULTS: The minimum inhibition concentration of A6, ACV and A6 + ACV measured by PCR was 1.88 mg/ml, 3.37 micrograms/ml and 0.47 mg/ml + 0.84 microgram/ml respectively, while measured by CPE, it was 6.25 mg/ml, 50 micrograms/ml and 0.94 mg/ml + 12.5 micrograms/ml respectively. The fractional inhibitory combined indexes were all less than 0.5, which is an indication of obvious co-synergistic effect. CONCLUSION: The quantitative PCR is an effective method of screening antiviral drugs. The inhibitory action of A6 and ACV on HSV1 revealed mainly at the replicative stage of viral proliferation cycle. PMID- 11475752 TI - [Exploration on personnel education of integrated Chinese traditional and Western medicine]. PMID- 11475751 TI - [An experimental study on curative effect of Chinese medicine qing yi tang in acute necrotizing pancreatitis]. AB - OBJECTIVE: To observe the curative effect of the traditional Chinese medicine (TCM) Qing Yi Tang (QYT) in acute necrotizing pancreatitis (ANP). METHODS: Twenty three dogs were randomly divided into 3 groups. In control group (n = 7), animals underwent laparotomy only. In ANP group (n = 8), acute necrotizing pancreatitis was induced by injection of 0.5 ml/kg 5% sodium taurocholate with 300 u/kg trypsin into the pancreatic duct. While in TCM group (n = 8) were fed everyday with QYT after onset of ANP. All animals were sacrified 7 days later and organs were gathered and cultured. Mucosal and luminal floras of the intestine were analysed. Pancreas and ileal mucosa were examined histologically and ultra microscopically, the levels of lipopolysaccharide (LPS) and amylase in blood were determined. RESULTS: In TCM groups, histologic and ultra-structural damages in pancreas and ileal mucosa were much milder as compared with those of ANP group. In ANP group, there was a significant increase of E. coli and bacterocoids, and a significant decrease of bifidobacteria, lactobacilli and enterococci in the intestinal mucosa, while in TCM groups, these changes were alleviated significantly (P < 0.05 or P < 0.01). As compared with ANP group, the bacterial translocation (BT) rate was reduced from 100% to 50%, and the counts of translocated bacteria were decreased 10-40 times, the levels of LPS and amylase reduced 2 to 3 times. CONCLUSION: TCM recipe QYT showed their protective effects on gut barrier function by alleviating the damage of intestinal mucosa and microecologic disturbance following acute pancreatitis. As a result, the chances of BT and enterogenic infection declined. These preparation might be promising in the prophylaxis and treatment of infection complicating ANP. PMID- 11475754 TI - Talking prescription labels available soon. PMID- 11475753 TI - [Current status of study on asthma treated with traditional Chinese medicine combined with Western medicine]. PMID- 11475755 TI - New drug for periodontal disease. PMID- 11475756 TI - Patients with PTSD risk damaging teeth. PMID- 11475757 TI - A study of Maryland dental hygienists' perceptions regarding self-assessment. AB - PURPOSE: This research applied the Theory of Reasoned Action to study dental hygienists' perceptions about self-assessment (SA). This theory suggests that individuals are likely to exhibit a given behavior, i.e., self-assessment, when they view it positively and when they believe that important others think they should perform it. Using the Theory of Reasoned Action model, the study addressed the following research questions: What are dental hygienists' beliefs, attitudes, motivations, and intentions regarding SA? Respondents were asked if they actually self-assess when providing dental hygiene services and if they felt they had adequate time to do so. The study also examined correlations between socio demographic variables (i.e., initial year of licensure, educational level, practice characteristics, and professional membership) and self-assessment intention and behavior. METHODS: A self-administered survey was mailed to 200 dental hygienists randomly selected from a list of all currently licensed dental hygienists in Maryland. Data analysis included descriptive statistics, correlations to predict intention and behavior, and analyses of differences related to socio-demographic variables. RESULTS: A total of 119 usable surveys were returned after one mailing, achieving a 59.5% response rate. The demographic data indicated that the respondents were a representative sample, with 66% holding associate degrees, 30% baccalaureate degrees, and 3% masters' degrees. Respondents' attitudes toward SA and intention to perform SA were strongly positive (X = 2.7, 2.6 respectively, on a Likert Scale ranging from -3 to +3) and, as the Theory of Reasoned Action suggests, attitude was strongly associated with intention (r = .6673). However, in contrast to the Theory, intention was weakly associated with behavior (r = .3721). Only one socio-demographic variable correlated with SA intention or self-reported behavior. Subjects who worked 20 hours per week or more were significantly more likely to self-assess when providing dental hygiene care than those who worked less than 20 hours per week (p < .01, .0089). A majority of respondents (76%) reported that they consistently perform SA and felt they had adequate time to do so. Respondents seemed to be motivated by perceived benefits of SA rather than by social pressure to perform it. CONCLUSION: Dental hygienists in this study appear to value self-assessment, use it when they provide patient care, and believe they have the time to do it. The extent and scope of their SA behaviors were not investigated. As SA is an integral component of quality assurance, application of well-developed self assessment strategies in both educational and practice settings may benefit both dental hygienists and the public they serve. PMID- 11475758 TI - Prevalence and risk factors of hand problems and carpal tunnel syndrome among dental hygienists. AB - PURPOSE: Recent research indicates a growing presence of hand problems and carpal tunnel syndrome (CTS) in dental professionals, especially among dental hygienists. This study was designed to determine the prevalence of hand problems and CTS among dental hygienists and identify risk factors for these conditions. METHODS: As part of a study that surveyed more than 5,000 army dental personnel, 177 dental hygienists were analyzed in great detail. Because of the magnitude of the overall study, which included all types of dental professionals, it was not only possible to identify the prevalence and risk factors of hand problems and CTS affecting dental hygienists, but also the prevalence rates as compared to other dental personnel. RESULTS: While the overall response rate for all dental personnel was 81%, dental hygienists responded at nearly 92%. Seventy-five percent of dental hygienists reported having hand problems, and 56% exhibited probable or classic symptoms of CTS. By logistic regression, the data revealed that dental hygienists whose practice comprised a majority (> 50%) of patients with heavy calculus were 2.3 times more likely to develop hand problems than those treating fewer patients with heavy calculus. Those who had practiced more than 10 years were also 1.9 times more likely to manifest symptoms associated with CTS than those with fewer years in the profession. CONCLUSION: The prevalence of hand problems and CTS among dental hygienists was the highest among army dental personnel, with the exception of dental therapy assistants. Risk factors for both hand problems and CTS are multifactorial, and dental hygienists should be particularly aware of those factors that can be prevented. PMID- 11475759 TI - Career retention in the dental hygiene workforce in Texas. AB - PURPOSE: The purpose of this study was to determine the factors that influence the retention and attrition of dental hygienists within the workforce in Texas. Respondents' perception of the role of employee benefits and practice of dental hygiene on career retention were explored. Demographic descriptors, including educational level, marital status, age, employment setting, and practice statuses, were also examined. METHODS: A questionnaire modified from the American Dental Hygienists' Association Extension Study: Retention of Dental Hygienists in the Workforce Final Report, April 1992, was mailed to a systematic sample of licensed Texas dental hygienists in March 1999. Descriptive statistics were computed for dental hygienists currently in practice in Texas and those not in practice at the time of the survey. Differences in demographics, benefits, and attitudes between dental hygienists currently in practice in Texas and dental hygienists not in practice at the time of the survey were tested using independent t-tests for interval data and chi-squared tests for categorical data. All statistical analyses were conducted using the Statistical Package for Social Scientists (SPSS v. 9, Chicago, Illinois). RESULTS: A response rate of 68.1% was obtained. Results revealed the primary reasons for remaining in the practice of dental hygiene were salary, family responsibility, professional collaboration, and variety of work. The primary reasons for leaving dental hygiene practice were family responsibility, boredom, salary, and lack of benefits. Secondary and tertiary reasons stated for staying in clinical practice revealed additional factors including benefits, participation in decision-making, and a safe environment. Dental hygienists in clinical practice were more likely to be employed by a dentist in a single practice and see more patients per day, have a certificate or associate's degree, be unmarried, have fewer children, and be younger than dental hygienists not in practice. CONCLUSION: The findings suggest that dental hygienists in Texas who remain in the workforce are positively influenced primarily by salary. Dental hygienists in Texas who had left the workforce were primarily influenced to leave practice because of family responsibility. Boredom and lack of benefits were also important factors in deciding to leave clinical practice. Employers of dental hygienists need to be aware of these factors in the hiring process. In addition, dental hygiene educators should prepare students in interviewing techniques for better communication regarding retention factors. PMID- 11475760 TI - The influence of toothbrush wear on the variables of plaque and gingivitis in clinical trials. AB - Although toothbrush wear has been shown not to be critical in ensuring optimal plaque removal, the degree of toothbrush wear occurring in a clinical trial may influence the results of the trial. The aim of this study was to determine the effects of toothbrush wear on the clinical variables of dental plaque and gingivitis. There were 107 subjects who participated in this six-month clinical trial in which plaque and gingivitis scores were assessed in relation to toothbrush wear at 2, 4, 12 and 24 weekly intervals. At each appointment, the toothbrushes being used were assessed for wear and replaced if necessary. It was found that 58% of the group were "rapid," and 42% were "slow" wearers. Of the group, 18.6% required replacement toothbrushes within two weeks from baseline with 50.9% of replacement toothbrushes being issued within four weeks of the start of the trial. Two subjects required as many as 10 replacement toothbrushes over the six-month trial period. "Slow" wearers had significantly lower plaque scores than "rapid" wearers at weeks 4 and 12, whereas the "rapid" wearers had significantly lower gingivitis scores than "slow" wearers at weeks 2 and 4. Clinical results show that significant toothbrush wear patterns of individuals participating in clinical trials should be taken into account when designing clinical trials assessing plaque and gingivitis scores. It may be necessary to exclude "rapid" toothbrush wearers from clinical trials that assess plaque and its removal. As the rate of toothbrush wear varied considerably among subjects, the simple classification of subjects into "rapid" and "slow" wearers may require further subgrouping. The surprising result that "rapid" wearers achieved lower levels of gingivitis requires further study to determine if this observation is valid in clinical trials designed for this purpose. PMID- 11475761 TI - Utilization of evidence-based teaching in U.S. dental hygiene curricula. AB - PURPOSE: The purpose of this research was to survey U.S. dental hygiene program directors to determine: (1) demographic information, (2) specific Evidence-Based (EB) student instruction methods used, (3) if and how programs use an EB philosophy, (4) perceptions of faculty skills in incorporating EB instruction, and (5) opinions and attitudes regarding future need to incorporate EB philosophies in dental hygiene education. METHODS: Data were gathered by surveying all 235 United States dental hygiene program directors in 1999. The survey included 20 closed items and 1 open-ended item. Initially, the survey was pilot tested using a convenience sample of seven U.S. dental hygiene program directors. A final, revised survey was mailed to the cohort population. A response rate of 70% (n = 164) was achieved after two mailings, and responses were analyzed using descriptive statistics. RESULTS: The demographic results of this study revealed the majority of respondents were from associate degree/certificate dental hygiene programs (77%). Results revealed that most dental hygiene programs are beginning to include some fundamental EB concepts and skills into their curriculum, primarily by incorporating analysis of scientific literature. Most programs provide students with formal library orientation (88%), instruct students in the use of library indices or library databases (86%), and teach the use of the Internet for conducting literature searches (79%). Respondents indicated the major barriers for fully incorporating an EB approach in their dental hygiene program were: lack of faculty skills (37%), no available time (34%), lack of financial resources (33%), and lack of technical support (28%). CONCLUSION: Findings of this study suggest dental hygiene educators have made small strides in creating an EB philosophy dental hygiene curriculum. However, the future of dental hygiene education must address the need for faculty development and training in areas such as computer utilization in core dental hygiene courses, strategies to improve the curriculum to stimulate students' critical thinking skills, and to develop educators' skills in the use of evidence for clinical decision-making. PMID- 11475762 TI - Practice behaviors of alumni trained as students in tobacco use cessation interventions. PMID- 11475763 TI - Automated oral hygiene self-care devices: making evidence-based choices to improve client outcomes. AB - Dental hygienists have historically been the primary promoters of health, prevention, and wellness in dental practice. With the large number of automated self-care devices available, many clients expect their dental hygienists to be experts and advisors on such purchases. It is vitally important for dental hygienists to understand and evaluate the available clinical research findings, and along with their clients, apply them in a collaborative decision making process. The purpose of this course is for dental hygienists to learn the scientific and practical implications of evidence-based care in the selection of automated oral hygiene self-care devices. Measurable clinical outcomes in plaque removal, reduction in gingivitis, bleeding, probing depth, periodontal pathogens, and the role of inflammatory mediators will be discussed. Automated self-care products currently available will be identified. Successful completion of the course objectives will be assessed by a post-test, to be completed by the learner after reading the article in its entirety. Two hours of continuing education course credit will be awarded following successful completion of the post test. PMID- 11475764 TI - Guidelines for the evaluation and management of traumatic dental injuries. PMID- 11475765 TI - Increased lifetime prevalence of dental trauma is associated with previous non dental injuries, mental distress and high alcohol consumption. AB - The purpose of the study was to assess the lifetime prevalence of dental injuries and risk factors involved in a general population-based birth cohort. The study population consisted of 5737 subjects who had participated in a health survey at the age of 31 years. Altogether 52% of the participants were women. This partly computer-based health survey included two questionnaires on previous dental and non-dental injuries, general health, occupational status and lifestyle. The current study was based on these questionnaires. The lifetime prevalence of dental fractures was 43% and the lifetime prevalence of dental luxations and exarticulations 14%. Men more commonly had dental injuries than women. Particularly mental distress and a history of previous injuries were shown to increase the risk for dental injuries. Furthermore, overweight and high alcohol consumption were positively associated with a high lifetime prevalence of tooth trauma. Regular physical activity decreased trauma occurrence. Socioeconomic status further affected the lifetime prevalence of dental injuries. The conclusion of the study was that personal, social and physical factors played a role in the occurrence of dental trauma. PMID- 11475766 TI - Social deprivation and traumatic dental injuries among 14-year-old schoolchildren in Newham, London. AB - AIMS: To assess the epidemiology of dental injuries among schoolchildren in Newham, London. MATERIAL AND METHODS: This study was carried out as part of the annual Community Dental Service's School Dental Inspection (SDI) programme in Newham. All schoolchildren aged 14 years were invited to participate in the study. Dental examinations were carried out by one trained examiner (SM) and included presence of clinical evidence of dental injuries, treatment provided and needed in the upper and lower permanent incisors, size of incisal overjet and type of lip coverage. The postcode was recorded to assess the levels of social deprivation. FINDINGS: 2242 out of 2684 children were examined. The prevalence of dental injuries was 23.7%. Treatment of dental injuries was neglected. Being male, having an incisal overjet greater than 5 mm and living in an overcrowded household increased the risk of having dental injuries. CONCLUSIONS: The prevalence of dental injuries in Newham (23.7%), which is a deprived area of London, was much higher than the overall prevalence in the UK (17%). Traumatic dental injuries seem to be a serious dental public health problem among children in deprived areas. Thus, there is an urgent need to collect local data on dental injuries in order to obtain a more comprehensive picture of the dental health. PMID- 11475767 TI - Prevalence and correlates of traumatic injuries to the permanent teeth of schoolchildren aged 9-14 years in Belo Horizonte, Brazil. AB - A cross-sectional survey was carried out on 3702 boys and girls aged 9-14 years, attending public and private primary schools in Belo Horizonte, Brazil. A multi stage sampling technique using an equal probability scheme was adopted to select the children. The response rate for the total sample was 97%. Dental examinations were carried out by one dentist (MISC). Intra-examiner agreement was very good. The prevalence of dental injuries increased from 8% at the age of 9 years to 13.6% at 12 and 16.1% at 14 years. Adjusted results showed that children from high socio-economic backgrounds were 1.4 (95% CI = 1.15-1.79) times more likely to present with a dental injury than children with low SES. Boys were 1.7 times (95% CI = 1.41-2.16) more likely to have dental injuries than girls. Children with an overjet size greater than 5.0 mm were 1.37 times (95% CI = 1.06-1.80) more likely to have a dental injury than children with an overjet size equal or lower than 5.0 mm. Finally, children with an adequate lip coverage were 0.56 times (95% CI = 0.44-0.72) less likely to have a traumatic dental injury than those with inadequate lip coverage. PMID- 11475768 TI - In vitro viability, mitogenicity and clonogenic capacities of periodontal ligament fibroblasts after storage in four media supplemented with growth factors. AB - The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to evaluate the effectiveness of growth factors (IGF-1 and PDGF-BB) when added to storage media in preserving the functional abilities of cultured periodontal ligament fibroblasts (PDLF). The evaluated storage media were: ViaSpan, Hanks' balanced salt solution (HBSS), alpha minimal essential medium (alpha MEM), and alpha MEM supplemented with FCS and antibiotic (alpha MEM-S). PDLF were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media supplemented with IGF-1 (10 ng/ml) and PDGF-BB (4 ng/ml) for 2, 8 and 24 h at room temperature (24 degrees C). The control group was incubated with the examined storage media without growth factors at 24 degrees C. An additional control group was incubated with culture medium at 37 degrees C without growth factors. After incubation, the viability of the cells was determined by Trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic (by culturing one cell/well) capacities. Storage of PDLF with growth factors (GF) for 2, 8 and 24 h decreased their vitality by only 3% (not statistically significant). The mitogenicity of PDLF stored for 2, 8 and 24 h in various media with GF was statistically comparable to that of the control group. Generally, the highest mitogenic capacity of PDLF stored with or without GF was found after 8 h of storage. Increasing the storage period to 24 h decreased the mitogenic capacity of the cells stored with GF by only 10-40% compared to the control group. In contrast, the clonogenic capacity of PDLF stored with GF increased with increasing storage periods by 100-300%, and the highest clonogenic capacity was found in most storage media after 24 h of storage with GF. The highest clonogenic and mitogenic capacities were found in cells stored in HBSS followed by alpha MEM-S. The mitogenic and clonogenic capacities of PDLF stored in various media supplemented with GF for 2-8 h were generally lower than without GF supplementation. The mitogenic and clonogenic effects of GF-supplementation was observed only after 24 h of storage. After 24 h of storage with GF, the clonogenic capacity increased by 8-224% and the mitogenicity by 20-37%, except in cells stored in alpha MEM (-1%). However, these differences were generally not statistically significant. In conclusion, the mitogenic and clonogenic effects of GF were observed only after 24 h of storage at room temperature. HBSS and alpha MEM-S supplemented with GF were the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for 24 h at room temperature. For short periods of storage (2 and 8 h), HBSS and alpha MEM-S without GF were preferable. PMID- 11475769 TI - Effect of enamel matrix derivative (EMDOGAIN) upon periodontal healing after replantation of permanent incisors in beagle dogs. AB - This study histologically assessed the effect of enamel matrix derivative (EMDOGAIN) on periodontal healing in re-implanted teeth in nine Beagle dogs. Incisors in dogs were re-implanted after 15, 30 and 60 min of dry storage with and without the application of EMDOGAIN. The pulps were removed in all teeth. In groups 1 and 2, dog teeth were splinted and the dogs sacrificed after 8 and 12 weeks, respectively. In group 3, dog teeth were not splinted and sacrifice took place after 12 weeks. The histological parameters studied were healed PDL, surface, inflammatory and replacement resorption. Univariable and multivariable analysis of data was performed. Increased incidence of healed PDL was inversely proportional to extraalveolar time. No significant differences were noticed between splinted and non-splinted teeth. The EMDOGAIN group showed a higher incidence of healed PDL, while controls showed a higher incidence of ankylosis. The effect of EMDOGAIN was more pronounced at the 12-week interval. PMID- 11475770 TI - Immediate repositioning of an accidentally extruded immature premolar after extracting root remnants of the primary molar. AB - This article describes a case where during an extraction of a primary molar's root remnant, the successor permanent tooth was accidentally extruded. The extruded tooth was immediately repositioned to its right position in the socket and it erupted normally after a short time. After 2 years, a routine clinical examination revealed normal appearance of the premolar in the dental arch. A radiograph demonstrated an obliterated coronal part of the pulp, normal root length, a slightly open apex and no periapical inflammation. PMID- 11475771 TI - Knowledge-based system for structured examination, diagnosis and therapy in treatment of traumatised teeth. AB - Dental trauma in children and adolescents is a common problem, and the prevalence of these injuries has increased in the last 10-20 years. A dental injury should always be considered an emergency and, thus, be treated immediately to relieve pain, facilitate reduction of displaced teeth, reconstruct lost hard tissue, and improve prognosis. Rational therapy depends upon a correct diagnosis, which can be achieved with the aid of various examination techniques. It must be understood that an incomplete examination can lead to inaccurate diagnosis and less successful treatment. Good knowledge of traumatology and models of treatments can also reduce stress and anxiety for both the patient and the dental team. Knowledge-based Systems (KBS) are a practical implementation of Artificial Intelligence. In complex domains which humans find difficult to understand, KBS can assist in making decisions and can also add knowledge. The aim of this paper is to describe the structure of a knowledge-based system for structured examination, diagnosis and therapy for traumatised primary and permanent teeth. A commercially available program was used as developmental tool for the programming (XpertRule, Attar, London, UK). The paper presents a model for a computerised decision support system for traumatology. PMID- 11475772 TI - Why epidemiological research in Pakistan? PMID- 11475773 TI - An objective structured practical examination to test students in experimental physiology. AB - OBJECTIVE: To develop a competency based discriminatory assessment method for physiology practical examination. METHOD: Results from 1st professional M.B.B.S. Part I and II of three batches were taken and students performance in traditional and objective structured practical examination (OSPE) were compared. The course objective for practical examination of all three batches were same. However, Batch II appeared in the conventional examination, while Batches III and IV were examined by OSPE. RESULTS: The mean score of Batch II was 68 +/- 6, of Batch III 53 +/- 13 and Batch IV 50 +/- 16. Batch II had thus an overall higher score as compared to Batches III and IV. The comparison of mean scores using ANOVA showed a significant (P < .001) difference between scores of Batch II as compared to Batches III and IV. Tukey's pair wise comparison of the batches showed a significant difference between batches II and III (95% CI for difference: 9.1, 20.5 with P < 0.001) and batches II and IV (95% CI for difference: 12.2, 23.6 with P < 0.001). However, no significant difference was found between batches III and IV (95% CI for difference: -2.6, 8.8 with P = 0.27). The result also showed that Batch II with conventional method of examination had a lesser spread around the mean (scores ranging from 52 to 81) as compared to Batch III (25 to 80) and Batch IV (14 to 90). CONCLUSION: OSPE is an effective tool to discriminate between good and poor performers in physiology practical examinations. PMID- 11475774 TI - P53 mutations in carcinoma breast--a clinicopathological study. AB - OBJECTIVES: To find out the incidence of p53 mutations and their possible correlation with clinicopathological presentation in females with breast carcinoma. SETTINGS: Department of Surgery, Pakistan Institute of Medical Sciences, Islamabad. PATIENTS AND METHODS: Seventy four patients with operable carcinoma breast that underwent mastectomy were included in this prospective study. Tumour tissue specimens and peripheral blood samples were examined for p53 mutations. Age, tumour size, nodal status and histopathology was assessed in patients with and without p53 mutations. RESULTS: Ten (13.5%) patients showed p53 mutations in their tumour specimens while 64 (86.48%) had normally functioning p53 gene. Patients were divided into two groups, A (normally functioning p53), and B (mutated p53). Intraductal carcinoma was the most frequent histological variant(A = 57, B = 10), while lymph nodes were involved in 67.19% (A = 47) and 60% (B = 6) cases respectively. The age of patients and clinical parameters (tumour size, nodal status and histopathological diagnosis) were compared between the two groups and no statistically significant correlation between p53 mutations and clinicopathological parameters was found. CONCLUSIONS: It was concluded that p53 mutation is present in carcinoma breast in Pakistani population but there was no significant correlation between p53 mutation and tumour aggressiveness (size, nodal status and histopathology). PMID- 11475775 TI - Blood stream infections in a medical intensive care unit: spectrum and antibiotic susceptibility pattern. AB - OBJECTIVE: To determine the type and sensitivity pattern of causative organisms of septicaemia in intensive care unit, to prepare a guideline for empirical antibiotic therapy. SETTING: Department of pathology and adult medical intensive care unit, PNS SHIFA (Naval Hospital), Karachi. METHODS: The study was conducted from January 1997 to June 1999. Blood specimens for culture were drawn from patients who developed symptoms/signs of bacteraemia/septicaemia 48 hours or more after admission in medical ICU. The specimens were inoculated into Brain Heart Infusion broth. Subcultures were done on days 1,2,3,5,7 and 10. The isolates were identified by standard biochemical tests. Antibiotic susceptibility pattern of the isolates was studied by Modified Kirby Baur method. RESULTS: Eighty-six aerobic organisms were isolated. They included Staphylococcus aureus(n = 34), Pseudomonas aeruginosa (n = 13), Escherichia coli and Enterobacter spp(n = 9 each), Klebsiella pneumoniae(n = 8), Acinetobacter spp and Serratia spp(n = 5 each), Citrobacter diversus(n = 2) and Proteus vulgaris(n = 1). On antibiotic susceptibility testing, 48.18% Staphylococcus aureus isolates were methicillin resistant. Susceptibility to other common drugs was also quite low while 100% of these were susceptible to vancomycin and amikacin. In case of gram negative rods more than 80% were resistant to ampicillin and cotrimoxazole. Susceptibility to gentamicin was as low as 25% for Klebsiella pneumoniae to 44.4% in case of Escherichia coli. Susceptibility to the third generation cephalosporins and the quinolone tested (ciprofloxacin) varied between 50-75%. All these isolates except Pseudomonas aeruginosa were susceptible to imipenem and amikacin. CONCLUSION: In view of the isolation of antibiotic resistant organisms, vancomycin in combination with amikacin or imipenem are the drugs of choice for empirically treating blood stream infections in ICU. Infection control procedures and antibiotic control policies can help to tackle this problem. PMID- 11475776 TI - Standard of knowledge about their disease among patients with diabetes in Karachi, Pakistan. AB - OBJECTIVE: The aim of this study was to determine the standard of knowledge among people with diabetes. METHOD: Two hundred and thirty patients were randomly chosen from outpatient clinics and a 34-item multiple choice questionnaire administered to them. The questionnaire was structured to assess knowledge about the disease state, diagnostic tests, complications and management. RESULTS: The average score of correct answers for the group was 40%. A significantly higher score correlated with younger age (16-30 years), educational status and regular follow-up with a diabetic clinic. There was no significant difference in the knowledge score between males and females or between those on oral hypoglycemic agents (OHA) and insulin. Fifty percent of the patients could correctly answers questions regarding food and nutrition and only 60% were aware of target blood glucose levels for optimal control. It was alarming to learn that 75% of those on insulin did not know that using U-40 or U-100 insulin does not change the dose required. CONCLUSION: The study emphasizes the need for diabetes education at all levels, both for the patients as well as the health care providers to counter the pandemic of diabetes-related complications globally. PMID- 11475777 TI - Role of traditional birth attendants in improving reproductive health: lessons from the family health project, Sindh. AB - OBJECTIVE: PURPOSE OF THE STUDY: Despite strenuous efforts, the maternal mortality rate in Pakistan remains high. The national figure of 340 maternal deaths per 100,000 live births tends to hide the fact that in some rural areas it is as high as 700 per 100,000 live births. Not surprisingly, in Pakistan only 20% of births are attended by a trained health professional. In most rural areas, home to almost 70% of the population, traditional birth attendants (TBAs) deliver 90% of the births. TBAs, therefore, play a crucial role in the delivery of maternal health care in Pakistan. Realizing the importance of TBAs, the Family Health Project (FHP) of the Department of Health Sindh, financed by the World Bank, tried to enhance their knowledge and skills through comprehensive training programs. FHP provided training to 650 TBAs in 10 districts. The training was provided by the Department of Community Health Sciences (CHS) of the Aga Khan University (AKU) who acted as technical consultant to the project. STUDY DESIGN: A community-based qualitative post-intervention survey. RESULTS: Post intervention survey of this seven-year project (1992-1999) revealed that (a) the training enhanced the knowledge and skills of the TBAs, (b) the trained TBAs provide more broader health care services and (c) they enjoy greater community acceptance and provide greater consumer satisfaction. It also showed that the TBAs remain the most available and accessible health resource in most rural settings. CONCLUSION: It is imperative that TBAs and their continuing training should remain central to any reproductive health intervention along with an effective referral system linking them to well-equipped emergency obstetric care facilities. However, the assessment clearly demonstrated that an integrated referral system backed by effective emergency obstetric care is essential to the success of the TBA training program. PMID- 11475778 TI - The effects of aerobic exercise training on resting blood pressure in hypertensive patients. AB - OBJECTIVE: To see the effects of aerobic exercise, on changes in blood pressure, in patients with essential hypertension. A 12-weeks aerobic exercise intervention trial was conducted, to examine the influence of brisk walking on resting systolic and diastolic blood pressure, pulse pressure, mean arterial blood pressure, body weight and body mass index in patients with essential hypertension. SUBJECTS AND METHODS: Twenty-seven men with stage 1 or 2 essential hypertension (not on antihypertensive medication) participated in the study. The aerobic exercise training protocol consisted of 30 minutes of brisk walking 3 to 5 times per week, at 50% of VO2max on an ergometer cycle. The data were analyzed by comparing exercise responses at baseline and 12-weeks. RESULTS: Statistically significant decrease in resting systolic [mean +/- SEM, 143.2 +/- 1.4 to 137.5 +/ 1.2 mmHg, mean reduction -5.7 mmHg, p < 0.01] and diastolic [mean +/- SEM, 91.8 +/- 1.0 to 90.4 +/- 0.9 mmHg, the mean reduction -1.4 mmHg] blood pressure were found (p < 0.05). Reduced pulse pressure from baseline value of -3.7 mmHg, (p < 0.01) and mean arterial pressure of -3.4 mmHg (p < 0.01) was noted. No discernible effects on mean body mass index was observed although mean body weights decreased -1.1 kg, (p < 0.05). Brisk walking yielded significant increase in VO2max (p < 0.05). CONCLUSION: Aerobic exercise caused small reductions in resting systolic and diastolic blood pressures in men with stage 1 or 2 essential hypertension. A lifestyle change such as exercising, may play a role in reducing the risk of hypertension. PMID- 11475779 TI - Social dimensions of child mental health in developing countries. PMID- 11475780 TI - Post-infarction left venticular free wall rupture. PMID- 11475781 TI - Anaesthetic management for a patient with severe multiple sclerosis. PMID- 11475782 TI - Physician assisted suicide perceptions among patients presenting to family physicians at a teaching hospital in Karachi, Pakistan. AB - OBJECTIVE: To study the perceptions on physician assisted suicide, among patients presenting to family physicians, at a teaching hospital in Karachi, Pakistan. METHOD: The study was carried out at the Community Health Center of The Aga Khan University Hospital, Karachi, between December 1999 and May 2000. The principal and co-investigators filled a pre-coded and pretested questionnaire consisting of important demographic characteristics and bio-medical ethics issues. A system of convenience sampling was used and a written consent was taken from respondents over the age of 16 years, who agreed to participate. The data were managed by using the Epi Info (version 6.0) program. RESULTS: Four hundred twenty respondents were interviewed against an estimated sample size of 385. Majority (88%) were males between 25 and 34 years of age, and were self employed or in private service. Overall 9% respondents were advocated of physician assisted suicide, advocates was 9%. Those who advocated the Physician assisted suicide were more likely to be female, elderly, married and educated amongst the total respondents. Those who support Physician assisted suicide were less likely to attach divine qualities to physicians and were more bold, courageous and tolerant towards broader biomedical ethics issues. CONCLUSION: We have found a substantial acceptability to the idea of Physician assisted suicide in a Muslim society and have identified characteristics of those who support it. PMID- 11475783 TI - [Beyond cholesterol levels: new therapeutic indications of statins]. AB - Cardiovascular diseases represent the major cause of morbility and mortality in industrialized countries, even if the incidence is quite different between each other. In the last years the knowledge of molecular mechanisms, which are responsible of progression of atherosclerosis vascular disease and of its complications such as coronary heart disease, has been particularly improved. At this regard new pharmacological compounds are introduced in clinical practice improved efficacy and effectiveness and, among these ones, thanks to statins a significative reduction in primary endpoint (i.e. reduction in incidence of stroke, total and cardiovascular mortality) has been already demonstrated. This effectiveness seems to be due by acting directly on the vascular atherosclerotic plaque independently from the reduction of blood cholesterol level which, on the contrary, may be involved in the progression of atherosclerotic vascular process. PMID- 11475784 TI - [Functions and changes of chemoreflex in physiological and pathological conditions]. AB - Chemoreceptors are known to respond to changes in composition of the gases in the inspired air by changing pulmonary ventilation. Whereas, less known it is the role that they play in many both physiologic and pathologic conditions. In this review we do not only consider the anatomic and physiologic chemoreceptor features but also we analyse the relationship between chemoreflex and baroreflex in the cardiovascular control, their role in the pathogenesis of essential hypertension and in the increase of blood pressure in subjects affected by obstructive sleep apnea, their implication in the genesis of dyspnea in chronic heart failure and their role in pulmonary disease. In this review we also analyse the chemoreflex changes with relation to physiologic situations such as aging, obesity, exercise, training and high altitude. Three methods are described for the study of the chemoreflex function, but more attention is paid to the new rebreathing technic. The aim of this paper is to underline the interactions between chemoreceptors and cardiovascular system and to analyse their functional changes in both physiologic and pathologic conditions. PMID- 11475785 TI - [The global village of celiac disease]. AB - Recent data suggest that celiac disease, or gluten-sensitive enteropathy, is a common disorder not only in populations of European ancestry, but also in developing areas, such as North Africa, Middle East and India. The world distribution of celiac disease apparently relates to the consumption of gluten containing cereals. In the Arab people of Saharawi, the prevalence of celiac disease in children is so high (more than 5%) that this condition represents a primary health problem. In developing countries the clinical picture of celiac disease is often typical, characterized by chronic diarrhea, malnutrition, stunting and increased mortality, especially in younger children. The high prevalence of celiac disease in areas showing a poor sanitary condition could be explained by a protective role of the celiac enteropathy (against intestinal infections) in the context of a low gluten consumption. The problem of celiac disease in developing countries should by approached by a multifaceted strategy, based on (1) increasing disease awareness and diagnostic facilities and (2) a "flexible" dietary intervention that takes into account the nutritional habits and local food availability. PMID- 11475786 TI - [Treatment with chimeric monoclonal antitumor necrosis factor (infliximab) of patients with active steroid-dependent/resistant Crohn's disease and fistulas]. AB - 30 patients--13 with active steroid-dependent/resistant Crohn's disease (CD), 8 with active steroid-dependent/resistant disease complicated by fistulas and 9 with fistulas only (perianal or abdominal)--were treated with infliximab. "Clinical response or remission" were defined as the reduction by 70 or more points or below 150 points of the CDAI score, respectively. As regards fistulas, "response" was defined as the reduction of 50 percent or more from baseline in the number of draining fistulas or of the quantity of drainage, "remission" as their closure. At 8 weeks 13/21 (61.9%) patients treated for active disease went on remission and 6/21 (28.5%) had a clinical response; 6/17 (35.2%) patients treated for fistulas went on remission and 8/17 (47%) had a response, while 3/17 (17.6%) didn't have any response. At 24 weeks, 9/12 (75%) patients treated for active disease and 13/16 (81.25%) treated for fistulas had a recurrence in a median time of 18.3 weeks (range, 1-36 weeks) after the first infusion. PMID- 11475787 TI - [Efficacy and safety of intravenously administered pantoprazole in the treatment of gastrinoma]. AB - The efficacy and safety of the proton pump inhibitor pantoprazole (P) in patients with Zollinger-Ellison syndrome (ZES) is well documented. We treated 21 patients with ZES with intravenous P; 13 of these patients were treated surgically in advance. The effect of P was very rapid, with reduction of the acid output within 15 min and its normalization within 60 min. The control of gastric acid secretion was maintained for a mean of 10.9 hours after the infusion of P. We conclude that the use of intravenous P allows a safe and efficient control of the acid ipersecretion in patients with ZES. PMID- 11475788 TI - [Lithium poisoning and the use of nimesulide]. AB - We describe a case of lithium intoxication, complicated by renal failure in a lithium treated women. She would take nimesulide, a selective cyclo-oxygenase-2 inhibitor, used in a variety of inflammatory, pain and fever states. This is the first report of the accumulation of lithium caused by nimesulide. PMID- 11475789 TI - [Immune restoration syndrome]. AB - A new syndrome, "immune restoration syndrome", has been described among patients with CMV retinitis, mycobacterial infections or cryptococcosis when HAART is initiated. The mechanism is most likely an enhanced immunologic reaction at the site of infection due to an improved T-lymphocyte function. The syndrome appears to be a rare event: the authors report a case of immune restoration syndrome in a patient with atypical mycobacteriosis. PMID- 11475790 TI - [Atypical presentation of lateral cervical metastasis of occult tumor]. AB - The authors describe the case of a 58 years old man, affected by severe obesity, alcohol addict, observed because of a retromandibular mass, measuring 6 cm. Clinical and instrumental findings indicated the presence of a mass apparently due to a parotid abscess; no evident lesions appeared at the examination of the ENT regions. Ultrasonography and fine needle aspiration cytology revealed the presence of a cystic cervical metastasis of a squamous cell carcinoma arising from an occult tonsillar cancer. The authors describe diagnostic procedures, differential diagnosis, treatment and peculiar aspects related to staging and follow-up. PMID- 11475791 TI - [Headache and coronary disease]. AB - Myocardial ischemia usually results in typical anginal chest pain, but cardiac pain can occasionally only be referred to regions outside the thorax. Headache, however, is a very unusual expression of myocardial ischemia. Thus, the correct diagnosis of "cardiac headache" is difficult and requires a high dose of suspicion, but it can be crucial to avoid possible life-threatening events. The type and site of headache are not specific, but an episodic occurrence. A short duration and a relationship with exercise or emotional triggers should alert physicians on the possible cardiac origin of headache. Paradoxical relief and prevention of headache by sublingual nitrates are a strong clue to the diagnosis. PMID- 11475792 TI - [Recent studies on the pathogenesis of cholelithiasis: the role of the gallbladder epithelium]. AB - The abnormalities of gallbladder epithelial function involved in cholesterol gallstone pathogenesis are: 1) the impaired biliary lipid absorption and 2) an increased secretion of mucin gel. We developed a model of isolated in vitro intra arterially perfused gallbladder and showed that the normal gallbladder epithelium absorbs high amounts of biliary cholesterol and phosphatidylcholine in a proportion determined by their molar ratio in the bile which enters the lumen. This physiological lipid absorption continuously reduces biliary cholesterol molar percentage in the gallbladder lumen and protects from gallstone formation by inhibiting cholesterol crystal precipitation. On the opposite, in the presence of cholesterol gallstone disease, even if the gallbladder epithelium is hyperplastic and hypertrophic, it absorbs cholesterol and phosphatidylcholine less efficiently and cholesterol crystal precipitation is favoured. Indirect evidence exists that the impaired gallbladder lipid absorption in cholesterol gallstone disease is not caused by inflammation and is not secondary to the presence of stones. The impaired mucosal function could be the consequence of excessive chronic enrichment of hepatic bile which enters the gallbladder. The mucin gel is the only biliary protein with an established pathogenetic role in cholesterol gallstone disease. The mucin gel is thought to favour gallstone formation by promoting cholesterol crystal precipitation and aggregation. PMID- 11475793 TI - [Endarterectomy in asymptomatic carotid stenosis: a controversial procedure for primary prevention of stroke]. AB - Stroke is the second cause of death and one of the main determinant of disability worldwide. Prevention remains the most important measure for reducing its incidence and mortality. Carotid endarterectomy is one of the possible means for reducing stroke incidence, at least in a subgroup of patients at risk of ischemic stroke. The benefit of carotid endarterectomy has been demonstrated consistently by two large international randomised controlled trials (North American Symptomatic Carotid Endarterectomy Trial, and European Carotid Surgery Trial) in patients with severe symptomatic carotid stenosis. In subjects with asymptomatic carotid stenosis the benefit has been shown by the only available trial (Asymptomatic Carotid Atherosclerosis Study), but proved to be quantitatively less relevant. The benefit is marginal if one considers that only part of stroke events occurring in the territory of an asymptomatic carotid stenosis is preventable by the intervention, since a consistent proportion (almost 50%) of these events are due to other causes (cardioembolism, lacunar infarction). Based on these observations, to obtain a beneficial effect, the perioperative risk should be minimized. Every surgical group should monitor carefully this risk, not omitting minor strokes, that in some cases are functionally relevant. Estimating prevalence and natural history of asymptomatic carotid stenosis in a definite population, the population impact number is expected to be very high, not justifying a widespread screening of the population aimed to identify subjects with asymptomatic carotid stenosis. PMID- 11475794 TI - [Hypochondroplasia: importance of radiological findings in the differential diagnosis of short statures of different origin]. AB - Hypochondroplasia is a genetic skeletal dysplasia characterized by disproportionate short stature, stocky appearance and several clinical and radiological features very similar, but much milder, than those of classical achondroplasia, including shortened and stubby long bones, decreased lumbo-sacral interpediculate distances, posterior scalloping of the lumbar vertebrae, metaphyseal flaring, and moderate macrocephaly. The condition may occasionally mimic short stature of familial, endocrine or metabolic origin. In the absence of clinical and laboratory diagnostic clues, radiological findings are of the utmost value in the diagnosis of this skeletal dysplasia and also in the differential diagnosis with other short-limbed dwarfisms. The genetic, clinical and radiological aspects of hypochondroplasia are briefly recalled, and the importance of some minor and frequently overlooked findings is stressed. PMID- 11475795 TI - [Nostalgia as a state of mind and as a clinical form]. AB - Psychiatry, as human science, necessarily concerns emotions too and, among these, nostalgia. Nostalgia is not only a mind condition but also a psychopathological and clinical state which can be defined nostalgic depression form. At the beginning of this work, on the base of a clinical case-report, we describe the phenomenological and space-temporal aspects of nostalgia, we analyse some psychopathological situations that, marked by nostalgic emotions, come from moving house, exile and rooting out. Each of these situations go with depression models thematically different, that are explained in the work. PMID- 11475796 TI - [Vascular effects of neurokinins in humans]. AB - Neurokinins (mainly substance P and neurokinin A) are released by sensitive nerve fibres. These fibres have been found in the vascular wall of arteries and veins of many vascular regions, particularly in nasal mucosa vessels, temporal and coronary arteries and saphenous veins. Substance P causes vascular relaxation by stimulating NK1 endothelial receptors. This relaxant effect is mediated, according to the vessels, by nitric oxide (NO), prostanoids or endothelium dependent hyperpolarizing factor (EDHF). Capsaicin, which induces the release of neurokinins, and neurokinin A can cause contractions of some vascular preparations, suggesting the existence of smooth muscle NK2 receptor associated with contraction. The vasodilatation induced by substance P injection appears reduced in patients with cardiovascular risk factors. The clinical development of specific neurokinin receptor antagonists may give the opportunity to specify the role of neurokinins in systemic vascular diseases. The results already obtained after repeated local applications of capsaicin (to reduce local levels of neurokinins) in vasomotor rhinitis and urticaria suggest that the vascular effects of neurokinins may participate in the clinical expression of these diseases. PMID- 11475797 TI - [Inhibition of uterine contractions: new in vitro pharmacological approaches on the pregnant human myometrium]. AB - The aim of this study was to evaluate the in vitro effects of phosphodiesterase 4 inhibitors (PDE4I) and their combination with salbutamol (beta 2-adrenoceptor agonist) on spontaneous contractions and to investigate by in vitro and biochemical studies and analysis of mRNA expression the presence of beta 3 adrenoceptor in human near-term myometrium. Rolipram, RP 73401 and Ro 20-1724 (PDE4I) inhibited spontaneous myometrial contractions (Emax approximately 100 per cent; pD2 approximately 6.80 for the two first and 6.31 for Ro 20-1724). Rolipram 10(-8) M potentiated the response to salbutamol (Emax = 88 per cent vs. 40 per cent and pD2 = 6.93 and 6.36 with or without rolipram respectively). SR 59119A, a beta 3-adrenoceptor agonist, was more efficient than salbutamol in inhibiting the contractions (Emax 52 per cent and 27 per cent respectively, p < 0.05) but they both induced a significant increase of cAMP production. In both functional and biochemical studies, SR 59119A was only antagonized by the beta 3-adrenoceptor antagonist SR 59230A. The beta 3-AR mRNA was positively expressed in myometrium preparations in a reverse transcription polymerase chain assay. In conclusion, phosphodiesterase 4 inhibitors alone or combined with beta 2-adrenoceptor agonists and beta 3-adrenoceptor agonists might have potential interest as tocolytic agents. PMID- 11475798 TI - The muscarinic receptor subtypes in human blood vessels. AB - Acetylcholine (ACh) may induce the relaxation and the contraction of human blood vessels. These effects involve the activation of muscarinic receptors located on endothelial or smooth muscle cells. In humans and animals, five subtypes of muscarinic receptors (M1-M5) have been identified. In the particular case of human blood vessels, the M3 subtype seems to be prevalent on endothelial cells as well as on smooth muscle cells. However, the M1 subtype may be specific for the pulmonary vascular endothelium. In contrast, the M4 subtype has not been described in human vessels. These conclusions are, however, based on a limited number of studies and further work is needed in this area of research. Although muscarinic receptors may not be the only receptors involved in hypertension, the development of specific agonists for M1 receptors may be useful to counteract any elevation of blood pressure in the pulmonary circulation. PMID- 11475799 TI - [Fom orphan receptor to novel neuropeptide: an example of reverse pharmacology]. AB - Nociceptin, a neuropeptide 17 amino acid residues in length, is the first novel bioactive substance to have been discovered by functional genomics. Nociceptin was isolated from a rat brain extract as the endogenous ligand to an orphan receptor, ORL1 (Opioid Receptor-Like 1), structurally akin to opioid receptors, whose cDNA had been cloned from a human brain stem library. The peptide is processed from a larger precursor polypeptide, prepro-nociceptin, which is widely distributed in the nervous system. Nociceptin is primarily an inhibitory neuropeptide that acts on neurons to depress synaptic transmission. Nociceptin's wide spectrum of pharmacological actions hints at several potential therapeutic applications for ORL1 receptor agonists and/or antagonists, particularly for treating pain states, stress and anxiety, cognitive defects, and drug addiction. A major challenge is now the discovery and pharmacological in vivo profiling of ORL1 receptor ligands that are active by medically practical routes of administration. PMID- 11475800 TI - [Functions of the 'Therapeutic Drug Monitoring (TDM)' Group of the French Society of Pharmacology]. AB - Therapeutic Drug Monitoring consists of sampling organization, measurement and interpretation of the blood concentration or biological effect of a given xenobiotic, and sometimes using this result for pharmacokinetic computation, with the aim of individual dose adjustment. This branch of pharmacology is represented internationally by the International Association of Therapeutic Drug Monitoring and Clinical Toxicology (IATDMCT) and has been organized, in France, as a working group of the French Society of Pharmacology (SFP). This group takes part in the main French organizations dealing with or using TDM, as well as the scientific activities of SFP and organizes multi-centre clinical trials studying exposure effect relationships or based on 'concentration-controlled' protocols. Finally, another aim of this TDM group is to promote the development and acknowledgement of this activity in hospitals. PMID- 11475801 TI - [Therapeutic drug monitoring (TDM): definitions--vocabulary and concepts]. AB - The French group 'Suivi Therapeutique Pharmacologique' (STP) has tried to define the best translation of the English term Therapeutic Drug Monitoring (TDM). The TDM concept was initiated with the development of Pharmacokinetics and Analytical Chemistry. At first, the aim was the adaptation to target concentrations of efficacy and toxicity. Then, monitoring became more individualized and took into account the patient, the indication and the coprescriptions. TDM participated in the improvement of knowledge of actors (physicians and biologists), the tools and the drugs concerned. The most recent developments have been the population approach, the Target Concentration Intervention (TCI) and the Randomised Concentration Controlled Trials (RCCT). TDM today is an individualized intervention that aims to achieve the best benefit/risk ratio, in the context of the patient and his/her entire environment. PMID- 11475802 TI - [Issues in therapeutic drug monitoring evaluation]. AB - A reassessment of the interest of therapeutic drug monitoring (TDM) is needed for a better definition of its applications. It rests on the hypothesis of the existence of an interindividual variability of the dose-effect relationship, this variability being influenced by pharmacokinetic variability. Different types of endpoints were used in validation studies (pharmacokinetic, indirect, economic) and few studies have used clinical endpoints. The question should be: does TDM allow for a better dose adjustment than one based only on criteria other than drug concentration? Retrospective studies have a 'diagnostic test' approach and only prospective clinical trials will provide a real validation. Factors which limit the setting up of such studies are: the number of subjects to include, the often imprecise measurement of drug exposure and poor knowledge of the pharmacokinetic-pharmacodynamic relationship and its variability. PMID- 11475803 TI - [Contribution of genotyping and phenotyping of biotransformation in therapeutic drug monitoring]. AB - Genetic-controlled biotransformation is a factor partly accounting for interindividual variability in drug responses. This control occurs via gene coding for polymorphic enzymes or rare genetic variants. Currently, the clinical relevance of genetically controlled biotransformation concerns only a few drugs. Determinants of clinical relevance depend on the size of the contribution of the metabolic pathway subject to genetic control, the existence of a relationship between the concentration and the therapeutic and/or toxic effect, a low therapeutic index and a pharmacokinetic variability smaller than pharmacodynamic one. Phenotyping using test drugs allows the determination of the activity of the drug-metabolizing enzyme but can be hampered by drug-drug interactions and some pathophysiological states. Genotyping can offer additional information such as the differentiation between ultra rapid metabolizers and patients who comply poorly with drug treatment, and the differentiation between genetic and environmental determinants. Genotyping and phenotyping can help predict dosage requirements. Currently the number of clinical laboratories using these tools in the drug-monitoring approach is small. PMID- 11475804 TI - [At which moment should one introduce Therapeutic Drug Monitoring into emergent therapy]. PMID- 11475805 TI - [Antiretroviral agents and pregnancy: mitochondrial dysfunction and nucleoside analogs]. AB - An alert was published during 1999 by the French Perinatal Cohort: eight cases of mitochondrial dysfunction were reported among 1754 infants exposed to nucleoside analogues in utero and during the neonatal period. These eight infants were not infected by HIV. Mitochondrial toxicity of nucleoside analogues is clearly described in adult HIV patients receiving NRTI. Zidovudine (the only and the first NRTI studied) induced mitochondrial DNA dysfunction in animals (monkeys) and neurobehavourial effects in mice at a dose similar to the human dose. Practitioners have been informed. Retrospective and prospective studies are in progress. The recommendations for prevention of maternofoetal transmission of HIV are not reassessed. Pregnant women in rich countries are receiving combination antiretroviral therapy. Information of women has to be undertaken and therapeutic strategies for maternal indication have to be discussed case by case. Careful long term follow up of children exposed to antiretroviral agents is a priority. PMID- 11475806 TI - [Drug interactions with antiretroviral agents]. AB - Concomitant administration of three or more antiretroviral drugs is the standard treatment for HIV-infected patients. I.p. and NNRT are metabolized by cytochrome P450 and are inhibitors or inducers of CYP3A4. Therefore a number of drug-drug interactions are likely to occur. Ritonavir, a potent CYP3A4 inhibitor, is coadministered with saquinavir, indinavir and amprenavir to enhance their plasma concentrations and their virological efficacy. In contrast, nevirapine and efavirenz are CYP3A4 inducers, which warrant an increase in i.p. dosing. These properties lead to interactions with other drugs metabolized by CYP3A4 and a knowledge or the route of biotransformation is useful to avoid side-effects or decrease efficacy (as in the case of statine coadministration). Some important interactions can lead to contraindications such as coadministration of rifampicine, astemizole, ergot derivates or cizapride, as a large decrease or increase in concentration can lead to inefficacy or to major side-effects. Clinical trials and notification of side-effects are important to detect unpredictable interactions and to propose guidelines; such an example is therapeutic drug monitoring of methadone to avoid withdrawal syndrome when coadministered with ritonavir or nelfinavir. PMID- 11475807 TI - [Prevention of adverse effects of drugs]. PMID- 11475808 TI - [Role of the pharmacist in the prevention of adverse drug effects]. PMID- 11475809 TI - The role of consumers. PMID- 11475810 TI - [Follow-up of 'medications and pregnancy' at the Midi-Pyrenees Pharmacovigilance Center']. AB - The follow-up of 283 gestations recorded by the Midi-Pyrenees Pharmacovigilance Centre concerned 544 medications, 17 solvents and 12 illicit drugs. Pregnancy outcomes were live births for 74.2 per cent of the women and voluntary (14.8 per cent), spontaneous (8.5 per cent) or therapeutic (1.8 per cent) abortions. Among the newborns, 11 had a malformation and 18 a neonatal disease. We observed an increased risk of neonatal disease for women exposed to illicit drugs during the third trimester of their pregnancy (OR = 14.28 [1.19-747.40] p = 0.0155) and for smokers (OR = 19.11 [2.11-445.00] p = 0.0011). Moreover, an increase of teratogenic risk was associated with a first trimester exposure to neurologic drugs (OR = 14.28 [2.08-100.00] p = 0.006). Women included in the present survey were more often exposed to potentially harmful drugs than the general population. However, the rates of obstetric and/or neonatal complications appear similar to those reported in France as a whole. PMID- 11475811 TI - [Adverse effects of Disulone; results of the France pharmacovigilance inquiry. Regional Centers of Pharmacovigilance]. AB - Disulone (dapsone + ferrous oxalate) is a sulphone marketed in France since 1958 and authorized in P. Carinii prophylaxis in HIV+ cotrimoxazole intolerant patients, bullous dermatosis, leprosy and polychondritis. Between 1983 and 1998, 249 adverse reactions were reported to French pharmacovigilance centres and Aventis, the manufacturer. Every side-effect was reviewed and the causal relationship was assessed on the basis of the French method for causality assessment. Main side-effects were divided as follows: 117 blood dyscrasias (generally neutropenia and agranulocytosis, rarely methaemoglobinaemia, haemolysis, macrocytosis, anaemia, aplastic anaemia, haemochromatosis and sulphaemoglobinaemia); 29 hypersensitivity syndrome; 39 cutaneous reactions, generally rash; 27 liver injuries (cholestatic, cytolytic and mixed hepatitis); 27 neurological and psychiatric side-effects including 7 axonal neuropathy; 10 gastrointestinal effects, generally nausea and vomiting. Five deaths were reported (4 septicaemia including one case not due to dapsone and 1 digestive bleeding due to underlying disease). In the other cases the outcome was favourable. The results were compared with the published references. It would seem to be important to reinforce information to prescribers about the possible serious adverse reactions with dapsone, particularly hypersensitivity syndrome and agranulocytosis, that can cause death if the drug is not stopped in time. PMID- 11475812 TI - [Inter-regional differences in dysthyroidism due to amiodarone: comparison of spontaneous notifications in Aquitaine, Midi-Pyrenees and Languedoc-Roussillon]. AB - Amiodarone hydrochloride is used in the treatment of ventricular and supraventricular arrhythmias. Because of its iodinated structure, thyroid dysfunction can occur during amiodarone therapy. The reported overall incidence is variable (2-24 per cent) and depends on several factors (past thyroid history, daily iodine intake,...). The present retrospective (1990-97) study was performed using the French pharmacovigilance database in order to compare the frequency of hypo- and hyperthyroidism in three areas in the South of France: Midi-Pyrenees, Aquitaine and Languedoc-Roussillon. For each case, the following data were recorded: age, sex, dysthyroidism history, dosage, duration and indication of amiodarone and delay to onset of dysthyroidism. We collected respectively 37, 50 and 9 cases of hypothyroidism in Midi-Pyrenees, Aquitaine and Languedoc Roussillon and 20, 69 and 11 cases of hyperthyroidism respectively in the same areas. These data show the predominance of reported amiodarone-induced hyperthyroidism in Aquitaine and Languedoc-Roussillon. Hypothyroidism seems more frequent in Midi-Pyrenees, a non-maritime area. The sex ratio (male/female) was significantly different for the occurrence of hypothyroidism in Midi-Pyrenees (1.8 versus 0.5 in Aquitaine and 0.8 in Languedoc-Roussillon). The delay to onset of hypothyroidism was significantly shorter in Midi-Pyrenees (17.1 months +/- 24.5) when compared with Aquitaine (28.7 +/- 28.1) or Languedoc-Roussillon (43.4 +/- 45). Our results show an interregional difference in the occurrence of hypo/hyperthyroidism due to amiodarone. PMID- 11475813 TI - [Drug evaluation in France: comparison of two approaches]. AB - Two independent expert groups assess new medicines in France: that of the journal Prescrire (P), and the official Transparency Commission (CT). Each uses its own 5 level scale to assess its contribution to therapeutics (P: (1) bravoi, (2) interesting, (3) adds something, (4) possibly useful, (5) nothing new; CT: (1) major, (2) important, (3) moderate, (4) modest, (5) none). We have compared the gradings for new drugs in P from 1993 to 1998 (except vaccines, generics and items for exclusive hospital use) to those of CT. Of 414 P gradings, 14 were without opinion, 54 were not found in CT (old or unexamined products), and 82 had been examined but not graded. The 264 graded pairs were 99 new substances (NS), 17 new combinations (NA), 50 new indications (NI), 56 new formulations (NF), 27 new dosages (ND), 10 new forms of packaging (NC), and 5 late gradings or reregardings (CR). For NS and NA, there were 21 discordances (2 degrees difference or more) between P and CT. When P is shifted one point to the left (better gradings), the number of discordances decreases to 2. NI, NF, ND, NC were less often graded by CT (respectively 63 per cent, 67 per cent, 54 per cent, 31 per cent of gradings). For NI, high CT grading results in 15 discordances (30 per cent). For NF, ND, NC the concordance is good (respectively 3, 1, 0 discordances). For CR, 13 of 18 products (72 per cent) are downgraded for P, and are discordant with the 5 initial CT grades. In 3 per cent of cases, P emits no opinion, CT gives a high grading in 2/3, never null (these are for severe diseases). In conclusion, the two groups of experts come to similar conclusions considering different objectives (use or not a new drug vs. give indications for reimbursement) and the time distortion. Discrepancies can also be explained thus, but some can result from differences in appreciation of the data. PMID- 11475814 TI - Inquiry into the distribution of drugs in a diabetic clinic. AB - The aim of this study was to investigate possible discrepancies between the drug prescribed and that recorded in the patient's file. A prospective open blind study was conducted with 178 patients included consecutively. We analysed 1011 prescriptions (the median (range) number of drugs per patient was 5 (1-37)) and identified 49 discrepancies (5 per cent of cases). In 18 cases, the drug given to the patient by a nurse was not the drug initially prescribed but the drug recorded in the patient's file was the drug actually given to the patient. In another 31 cases, the drug given to the patient was not the drug initially prescribed, but the drug recorded was that prescribed. This inquiry shows that there may be a discrepancy between the drug initially prescribed and that actually administrated and suggests that poor traceability may affect pharmacological surveillance surveys. PMID- 11475815 TI - [Interaction between melilot and acenocoumarol? (Melilotruscus aculeatus)]. PMID- 11475816 TI - [Cerebral meningeal hemorrhage and acute cerebral angiopathy associated with the taking of phenylpropanolamine: a new case]. PMID- 11475817 TI - ICD product reliability: lessons from electrical overstress failures. PMID- 11475818 TI - Unexpected ICD pulse generator failure due to electronic circuit damage caused by electrical overstress. AB - Because it is a lifesaving device, the unexpected failure of an ICD can be catastrophic. We report ICD electronic circuit failure due to electrical overstress damage (EOS) to the high voltage hybird circuit and other electronic components in a series of ICD pulse generator models. Data were obtained from the Multicenter Registry of Pacemaker and ICD Pacemaker and Lead Failures, and from the manufactures' adverse event reports, that were in the FDA's Manufacturer and User Facility Device Experience (MAUDE) database. Of 16 nonbattery Guidant/CPI ICD pulse generator failures reported to the registry, 6 (38%) have been confirmed by the manufacturer to be EOS related, and Guidant/CPI has reported 273 such failures to the FDA as of 12/29/00. The signs of failure included loss of telemetry and inability to deliver therapy, and some patients have experienced serious adverse events. Hybrid circuit damage may have occurred during capacitor charging or reform, and the majority appears to have happened during normal ICD function. While the incidence of this problem is unknown, a management strategy should be adopted that includes routine follow-up every 3 months and device evaluation after a shock or exposure to external defibrillation or electrosurgical devices. This study suggests that additional data are needed to determine the incidence of this problem, and that our present methods for monitoring the performance of ICD's following market release are inadequate. PMID- 11475819 TI - Intraoperative study of polarization and evoked response signals in different endocardial electrode designs. AB - Some new generation pacemakers use an algorithm based on evoked response (ER) detection to verify beat-to-beat capture and to enable automatic adjustment of output. This is a prospective acute study of polarization signal (PS) and ER in nine currently available electrodes. Intraoperative testing of ventricular bipolar electrodes used the Autocapture (AC) algorithm. The intrinsic R wave, PS, ER, acceptance of AC function, and stimulation thresholds (STs) were obtained. Ventricular electrodes were categorized as follows: titanium nitride (TiN)-coated passive and active fixation, high impedance (HI), passive fixation (VP), iridium oxide-coated titanium (IROX) (VI), and platinum helix (PH) active fixation. Acute testing was performed in 217 patients with an average age of 74.26 years, 59.6% were men with primary pacing indication-SSS (46.3%). There were no significant differences found with respect to R wave and threshold between the various electrodes. PH active-fixation electrodes had significantly higher ER and PS than other groups including the TiN-coated active-fixation electrodes. TiN-coated electrodes (active and passive fixation) had significantly lower PS than other electrodes. As a result, TiN electrodes had a significantly higher functional rate of AC (91.7%), whereas PH had the lowest rate (0%). In conclusion, (1) polarization characteristics are significantly different for commercially available ventricular electrodes, (2) certain physical features at the tissue to electrode interface like TiN coating appears to be more important in determining PS than electrode tip size and fixation method, and (3) the current algorithm for AC requires electrodes that provide low polarization for satisfactory performance. PMID- 11475820 TI - Prospective evaluation of catheter ablation in patients with implantable cardioverter defibrillators and multiple inappropriate ICD therapies due to atrial fibrillation and type I atrial flutter. AB - The high incidence of inappropriate therapies due to drug refractory supraventricular tachycardia remains a major unsolved problem of the ICD. Most of the inappropriate therapies for supraventricular tachycardia are caused by AF and type I atrial flutter with rapid ventricular response. The purpose of this prospective study was to determine the usefulness of AVN modulation or ablation for rapid AF and ablation of the tricuspid annulus-inferior vena cava (TA-IVC) isthmus for type I atrial flutter in ICD patients with frequent inappropriate ICD interventions. Eighteen consecutive patients were enrolled in this study. Twelve patients received a mean of 34 +/- 36 antitachycardia pacing (ATP) and 41 +/- 32 shock therapies for rapid AF during 49 +/- 39 months, and 6 patients a mean of 111 +/- 200 ATP and 11 +/- 8 shock therapies for type I atrial flutter during 52 +/- 37 months preceding ablation procedure. Modification of the AVN was successful in 10 (83%) of 12 AF patients, in 2 (17%) patients ablation of the AVN was performed. A complete TA-IVC isthmus block was achieved in 5 (83%) of 6 atrial flutter patients. Three (25%) AF patients had 11 +/- 24 recurrences of ATP and 0.4 +/- 1.1 shock therapies for rapid AF during 15 +/- 7 months. None of the atrial flutter patients had recurrences of inappropriate therapies for type I atrial flutter during 14 +/- 8 months, but two (33%) patients had inappropriate ICD therapies for type II atrial flutter or rapid AF. There was an overall mean incidence of 18 +/- 22 inappropriate ICD therapies per 6 months before and 4 +/- 9 per 6 months after the ablation procedure (P < 0.05). In conclusion, radiofrequency catheter modification or ablation of the AVN for rapid AF and ablation for atrial flutter type I are demonstrated to be highly effective in the majority of ICD patients with drug refractory multiple inappropriate ICD therapies. PMID- 11475821 TI - Dispersion in ventricular repolarization in patients with severe intraventricular conduction disturbances. AB - Increased dispersion of repolarization, measured invasively or by QT interval measurements, is associated with an increased risk for ventricular arrhythmias and sudden death. Most studies on this issue have included patients with normal intraventricular conduction, and it is not known if this finding has a predictive value also in patients with intraventricular conduction disorders. An invasive electrophysiological study, including programmed ventricular stimulation and assessment of effective refractory periods at two RV sites, was performed in 103 patients with bifascicular block (mean age 67 +/- 12 years). QT dispersion was measured from standard 12-lead ECGs. In patients with inducible sustained polymorphic VT or VF the dispersion in refractoriness between the two RV sites was significantly greater (46 +/- 11 ms, n = 13) than in noninducible patients (14 +/- 14 ms, n = 84) and in patients with inducible sustained monomorphic VT (16 +/- 5 ms, n = 6) (P < 0.01). Similarly, QT dispersion was 104 +/- 46 ms, 66 +/- 31 ms, and 77 +/- 33 ms, respectively, in the three groups (P < 0.05). Dispersion in repolarization, neither measured invasively nor by QT interval measurements, predicted sudden death, all cause mortality, or ventricular arrhythmia during a mean follow-up period of 3 years. In patients with bifascicular block, there is a relation between the degree of dispersion of ventricular repolarization and the inducibility of polymorphic ventricular arrhythmia, but this outcome did not occur during follow-up. PMID- 11475822 TI - The prognostic significance of atrial arrhythmias recorded early after cardioversion for atrial fibrillation. AB - In a substantial number of patients, AF recurs after successful electrical cardioversion. The purpose of this study was to investigate if the atrial arrhythmias recorded immediately after cardioversion are associated with the risk of recurrence of the arrhythmia and to compare the prognostic significance of this parameter with that of other established risk factors. In a series of 71 patients, the risk factors for recurrence of AF during the first year after successful electrical cardioversion were analyzed. A new parameter that was investigated was the frequency of atrial premature beats and the presence of runs of supraventricular tachycardia in the Holter recording started immediately after the cardioversion. Age, left atrial size, left ventricular systolic function, duration of the arrhythmia before cardioversion, underlying cardiac disease, or medication taken were not found to be predictive of recurrence of the arrhythmia. However, the natural logarithm of the number of atrial premature complexes per hour of the Holter recording in the 37 patients in whom AF recurred was higher compared to that of the 34 patients who maintained sinus rhythm (P < 0.0005). The same was true if only the first 6 hours of the recording were analyzed (P < 0.0005). There was a trend for more frequent arrhythmia recurrence if runs of supraventricular tachycardia were present. The finding of > 10 atrial premature complexes per hour in the recording had a relative risk of 2.57 (1.51-4.37), a positive predictive accuracy of 76.5%, and a negative predictive accuracy of 70.3% for subsequent arrhythmia recurrence. We can conclude that frequent (> 10/hour) atrial premature complexes in the Holter recording after electrical cardioversion for AF is a significant risk factor for recurrence of the arrhythmia. PMID- 11475823 TI - Mapping the organization of atrial fibrillation with basket catheters. Part I: Validation of a real-time algorithm. AB - The authors proposed a real-time algorithm to quantify AF organization from multielectrode basket catheter (MBC) recordings. The algorithm is based on a fast method to estimate the number of points along the baseline of a single bipolar electrogram (number of occurrences, NO). They have already proven this parameter to be effective at discriminating AF organization according to Wells' criteria. Special attention has been paid to ventricular far-field artifacts, as they proved they can significantly bias the degree of organization. To fulfill the constraints of a real-time application, a ventricular blanking procedure was implemented and validated. Bipolar electrograms were obtained from MBC in the right atrium in 17 informed patients with chronic AF. The results of NO computations were displayed as three-dimensional color-coded maps of organization by interpolating the measurements obtained at the locations of catheter bipoles. The proposed method allows real-time estimation of the organization of the atrial electrograms according to Wells' criteria. The estimation has a temporal resolution of 2 seconds, is robust to far-field ventricular artifacts, and interpolates the data available to furnish a single global map of the entire atrium. Further studies devoted to the analysis of individual and common patterns of the regional distribution of AF organization are needed to assess the usefulness of this approach for electrophysiological studies and therapeutic applications. PMID- 11475824 TI - Mapping the organization of atrial fibrillation with basket catheters. Part II: Regional patterns in chronic patients. AB - The aim of this study was to investigate the spatial patterns of organization of fibrillation in the right atrium using bipolar recordings from a basket catheter in a population of 17 patients with persistent AF. The number of occurrences (NO) (i.e., the number of points laying on the baseline) was used as an objective measure of organization. This parameter significantly matches Wells' criteria for the classification of bipolar recordings during AF. The anatomic correspondence of the splines with eight atrial regions (namely anterior free wall, lateral free wall, posterolateral free wall, posterior free wall, posterior atrial septum, atrial septum, tricuspid valve, and anterior tricuspid valve) was assessed by fluoroscopic projections and electrogram morphology. Each region was further divided into high, mid-high, mid-low, and low segments. It was observed that individual and well-defined patterns of organization characterize the electrical activity of the right atrium in patients with persistent AF. When observed over 10 minutes, these patterns are rather stable in time. In each patient, the NO measurements were significantly different in the various right atrium regions (P < 0.0001 for all patients, Friedman test). The NO averaged over the population was significantly different among the regions (P < 0.001, Kruskal-Wallis test), owing to a common disorganized pattern observed in the tricuspid valve and anterior tricuspid valve regions. When these regions were removed from the analysis, the differences among the remaining regions were not statistically significant (P = 0.77). In conclusion, the quantitative assessment of organization in the entire right atrium showed an individual rather than common distribution. This may have implications in the choice of regions candidate for ablation. PMID- 11475825 TI - Noninvasive diagnosis of a wide QRS complex tachycardia mechanism by programmed atrial and ventricular stimulation using a dual chamber defibrillator. AB - Stored atrial and ventricular electrograms retrieved from dual chamber implantable defibrillators facilitate the diagnosis of arrhythmias. This case also illustrates the usefulness of programmed atrial and ventricular stimulation for noninvasive rhythm diagnosis in patients with a wide QRS tachycardia and an implantable defibrillator. PMID- 11475826 TI - Effect of procainamide on the postrepolarization refractoriness in cardiac muscle: evaluation using the block coupling interval in the artificial isthmus model in the canine right atrium. AB - The post-repolarization refractoriness (PRR) is an important factor to determine the conduction block in cardiac muscle. Recently, we proposed the block coupling interval (BCI) as an useful electrophysiological index for evaluating the PRR. In the present study, the effect of procainamide on PRR was evaluated using the BCI and the effective refractory period (ERP). In five beagle dogs, radiofrequency linear ablation was performed on the right atrial surface parallel to the AV groove, forming an artificial isthmus (8-10 mm width and 15-20 mm length). Bipolar recordings were performed in the isthmus at a resolution of 1.2 mm and single extrastimuli with eight basic drive trains were delivered to cause conduction blocks in the isthmus. When a conduction block occurred, the recorded coupling interval at the recording site just proximal to the site of block was defined as BCI. At the site of the block, the ERP and duration of the monophasic action potential (MAP) at each drive cycle length was measured. The PRR was calculated using two different formulas: (1) [ERP-MAP] and (2) [BCI-MAP]. Procainamide was administrated intravenously at a dose of 15 mg/kg after the control study and the whole study protocol was repeated. The site of the block in an individual dog was always the same. BCI, ERP, and MAP were all shortened in accordance with the shortening of the basic drive cycle length, and the BCI was always the longest, ERP the middle, and the MAP was the shortest. The administration of procainamide prolonged each parameter, but the order of BCI > ERP > MAP remained unchanged. The PRR calculated as [BCI-MAP] was prolonged from 15 +/- 10 ms to 29 +/- 8 ms by the administration of procainamide (P = 0.048), but [ERP-MAP] was unchanged (8 +/- 10 ms vs 8 +/- 4 ms). In the conduction block model in the canine right atrium, procainamide prolonged the [BCI-MAP], but did not change the [ERP-MAP]. The procainamide effect of prolonging the PRR might be expressed better by the change in the BCI than the ERP. PMID- 11475827 TI - Internal low energy cardioversion of atrial fibrillation using a single lead system: comparison of a left and right pulmonary artery catheter approach. AB - Internal cardioversion (ICV) has been demonstrated as an effective and safe method for restoring sinus rhythm in patients with AF. Recently, a new single lead system with a balloon-guided cardioversion catheter was introduced. ICV was performed after advancing a 7.5 Fr catheter flow-directed into the left or right pulmonary artery (PA, distal array, cathode). The proximal array (anode) was placed at the lateral RA wall. Synchronized shocks (3/3 ms biphasic impulse) were applied using a stepwise protocol (0.5, 3, 6, 9, 12, 15 J) until sinus rhythm was restored or maximum energy (15 J) was reached. Sixty-five patients (mean age 58 +/- 13 years) with acute and chronic AF were included. Sinus rhythm could be restored in 59 (91%) patients. Cardioversion success was 93% in the left PA compared to 86% in right PA. DFTs for the left and right PA approaches were 7.1 +/- 4.0 J and 10.2 +/- 4.0 J, respectively (P < 0.0001). It was significantly higher in patients with an AF history > 7 days (7.2 +/- 4.1 J) than for those with a recent onset of AF (5.6 +/- 4.1 J), P = 0.0012. Shock impedance differed for the left and right PA lead configuration (53 +/- 11 vs 49 +/- 13 omega, P < 0.05). A right PA lead configuration is as effective compared to a left PA catheter approach when performing ICV for AF. ICV with a single lead system is safe and cardioversion success is comparable to other internal and external cardioversion techniques. In combination with hemodynamic monitoring, flow directed nonfluoroscopic catheter positioning is feasible and may serve as a valuable therapeutic and diagnostic tool in intensive care units. PMID- 11475828 TI - Young at heart: understanding the unique psychosocial adjustment of young implantable cardioverter defibrillator recipients. AB - This article reviews the data related to psychosocial adjustment of young ICD recipients, postulates theories to explain potential adjustment difficulties to ICD therapy experienced by younger recipients, and suggests clinical management techniques for addressing the unique psychosocial concerns of young ICD recipients. Studies of young ICD recipients suggest that a wide range of psychosocial adjustment issues are prominent in the post-ICD implantation period and that the issues may be different from older ICD recipients. The disability stress-coping model and the transactional-stress-coping model are postulated as explanations for the unique adjustment concerns of children and adolescents with ICDs. Social comparison theory is also applied to the concerns of young adults with ICDs such that they often lack same age peers to compare experiences with cardiac difficulties. Brief, clinic-based interventions by health care providers, like a screening and referral heuristic and an "ICD Buddy" system, are suggested to increase effective coping and decrease social isolation for young ICD recipients. PMID- 11475829 TI - Comparison of two diagnostic algorithms for regular broad complex tachycardia by decision theory analysis. AB - Sensitivity and specificity are two inversely related properties of a diagnostic test and it is often practically infeasible to secure a high value for both simultaneously. Decision theory analysis shows that the utility of a diagnostic test depends not only on its sensitivity and specificity but also on the prevalence of the intended target disorder: when prevalence is low, a high specificity is more important than a high sensitivity, whereas when prevalence is high, a high sensitivity is more important than a high specificity. The significance of this principle is illustrated by two popular algorithms for the electrocardiographic diagnosis of regular broad complex tachycardia (BCT), of which the two main differential diagnoses are ventricular tachycardia (VT) and supraventricular tachycardia with aberrant conduction (SVTAG). Brugada et al. focused on criteria highly specific for VT and used them to build a four-step algorithm. In contrast, Griffith et al. first selected criteria highly sensitive for VT and then criteria highly specific for VT to build a simple two-step algorithm. It can be objectively demonstrated that the Griffith algorithm is more efficient and effective than the Brugada algorithm in terms of clinching the final diagnosis and improving overall diagnostic accuracy. The main reason for this is that VT is more common than SVTAC as the cause of regular BCT, and the Griffith algorithm adhered to the aforementioned principle governing the choice between sensitivity and specificity according to prevalence in its design. The Griffith algorithm also embodies an additional important principle, namely, it is easier and more efficient to choose alternatively between criteria highly specific and highly sensitive for the intended target disorder than concentrating on just one or the other in designing a multiple-step sequential diagnostic algorithm. PMID- 11475830 TI - The mysterious fate of Hyman's pacemaker. PMID- 11475831 TI - The ablation of atrial fibrillation with the loop catheter design: what we have learned from the animal model. AB - Ablation of chronic atrial fibrillation (AF) with the use of transcutaneous catheter ablation is yet to become a clinical tool. This article summarizes the development of the technology and the technique for the ablation of AF that was tested on the rapidly paced AF dog model. The current ablation technology using the standard ablation technique used in humans is not suitable for the creation of transmural contiguous linear lesions, and such technology is subjecting the patient to the prolonged procedures with considerable risk of complication. The use of the loop catheter design or other designs of ablation technology, which is specifically targeted for the creation of linear lesions, should be developed if the catheter approach for the ablation of AF is to succeed. This article describes the data available for the loop catheter design that is currently undergoing human feasibility studies. PMID- 11475832 TI - A regular narrow QRS complex tachycardia with atrioventricular dissociation. PMID- 11475833 TI - Inappropriate shock delivery due to ventricular double detection with a biventricular pacing implantable cardioverter defibrillator. AB - This report describes a patient with advanced heart failure, pronounced intraventricular conduction delay, and ventricular tachycardias who underwent implantation of a multisite pacing ICD. Pacing leads were placed in the right atrium, right ventricular apex, and to the left ventricular posterior wall via a coronary sinus vein. The system proved to have correct sensing and pacing function in an atrial synchronized biventricular pacing mode and an appropriate detection of ventricular fibrillation. However, 1 month after implantation the patient received an inappropriate shock delivery due to double detection of ventricular premature beats. The inherent detection problem of dual ventricular sensing is discussed. PMID- 11475834 TI - Spontaneous regression over a 16-year period of tachyarrhythmias to sick sinus syndrome and complete atrioventricular block in a young patient with Ebstein's anomaly. AB - A 25-year-old man with Ebstein's anomaly showed spontaneous regression of tachyarrhythmias to sick sinus syndrome and complete atrioventricular block over a 16-year period. This is the first clinical report supporting the hypothesis that abnormal cell death might contribute to the disturbance of the heart conduction system in Ebstein's anomaly. PMID- 11475835 TI - Cardiac leiomyosarcoma of the right atrium in a teenager: unusual manifestation with a lifetime history of atrial ectopic tachycardia. AB - A 16-year-old girl presented with atrial fibrillation. Transesophageal echocardiography revealed a right atrial leiomyosarcoma. Her past medical history was remarkable for incessant atrial ectopic tachycardia (AET) beginning in early infancy and continuing throughout childhood and adolescence that was refractive to medical and nonpharmacological treatment. After combined surgical and medical therapy, normal sinus rhythm was restored and the patient is currently in complete remission with no recurrent symptoms or atrial arrhythmias at 31 months after surgery and 23 months after the discontinuation of chemotherapy. Atrial tachycardia may be the first, and for prolonged periods, the only manifestation of a cardiac tumor and should prompt thorough investigation of its underlying morphological substrate. PMID- 11475836 TI - Triple-site ventricular pacing in a biventricular pacing system. AB - This case report describes a patient with heart failure in whom a biventricular pacing system was successfully implanted. During control of the pacing system, three morphologies of the paced QRS complex could be elucidated. Right ventricular stimulation, biventricular stimulation, and biventricular pacing with additional stimulation from the anodal electrode of the right ventricular lead determined the morphologies. PMID- 11475837 TI - Unexpectedly high defibrillation thresholds in an active can implantable cardioverter defibrillator system. AB - This case report describes a patient undergoing defibrillator generator replacement in whom the defibrillation thresholds were significantly higher with a can-active system than with a non-active can system. PMID- 11475838 TI - Acute loss of capture due to flecainide acetate. AB - Antiarrhythmic drugs increase pacing threshold, but this is rarely of clinical significance. Administration of flecainide acetate in a 75-year-old woman with an implanted AAIR pacemaker because of sick sinus syndrome caused an abrupt rise of pacing threshold and failure of pacing. Pacing threshold returned to the normal value a few days after flecainide treatment was stopped. PMID- 11475839 TI - Anticoagulation reduces strokes in atrial fibrillation (AF). PMID- 11475840 TI - Long-term experience with the electrogram (EGM) width criterion for differential of supraventricular tachycardia (SVT) and ventricular tachycardia (VT) in patients with implantable cardioverter defibrillators (ICDs). PMID- 11475841 TI - [Diagnostic difficulties with differentiating noninfectious diseases with coexisting cerebrospinal fluid changes--personal observations from the Department of Infectious Diseases, Jagiellonian University in Cracow]. AB - We present two cases of women admitted to our department with suspicion of meningitis. They were transferred from Neurology Unit. CSF changes found in both cases were not of infectious origin. Authors pay attention to diagnostic difficulties in such cases. PMID- 11475842 TI - [Neurofibroma of the small intestine as a cause of massive bleeding from the gastrointestinal tract]. AB - Two cases of neoplastic tumors of small intestine, operated on because of acute abdominal signs, were described. In both cases, before ultimate surgical treatment, chronic gastro-intestinal bleeding and ileus were observed. None of routine diagnostic procedures (endoscopy and angiography) diagnosed the disease. It was the radiological contrast examination of small bowel, that solved the problem. For the reason, that neoplastic tumors of small intestine are extremely rare and symptoms are not specific, results of treatment were described. Also the mistakes in diagnostic process were analysed. Malignant lesions are very rare and they are not symptomatic, those are the reasons of delayed surgical treatment. Moreover, they are more common in older age (over 60), that makes the invasive diagnostics even more difficult because of general condition and additional loads, e.g. atherosclerosis or circulatory and respiratory failure. Literature survey underlines the fact, that difficulties in the diagnostics process lead to developing of advanced stages of neoplastics disease, which are treated because of massive bleeding ileus. Often patients are hospitalized many times before the ultimate surgery, but are discharged without right diagnosis. Therefore the retrospective analysis of our cases can be useful in future for the proper interpretation of clinical signs. PMID- 11475843 TI - [Evaluation of efficacy and toxicity of docetaxel (Taxotere) in patients with advanced mammary gland cancer]. AB - Twenty women with advanced breast cancer were treated with Docetaxel. In 50% of cases partial remission with median duration of 7.1 months was obtained. The median survival time was 18 months. Stabilization of disease with median duration of 5.5 months was obtained in 45%. The median survival in this group of patients was 15.3 months. The pervious antracycline-based chemotherapy did not influenced the results. The most common side effect was neutropenia (G3 in 80% of pts) and alopecia, but the chemotherapy tolerance was satisfactory. PMID- 11475844 TI - [Modeling long-term results of basiliximab (SIMULECT) use in patients after kidney transplantation in Great Britain]. AB - Kidney graft survival prognosis could be performed in relation to the number of acute rejection episodes and their severity. The episodes of rejection could be modelled and based and the results obtained on this way could serve for prediction of a length of graft survival as well as for basiliximab treatment planning. PMID- 11475845 TI - [Serine protease inhibitors in plasma of patients with ulcerative colitis]. AB - Serine protease inhibitors (serpins) are the important factors regulating hemostasis and inflammatory mediators activity. The aim of this study was to determine relationships between plasma serine protease inhibitors activities and the course of ulcerative colitis (uc). PATIENTS AND METHODS: 42 patients with uc, 21 in active and 21 in inactive disease phase, and 26 healthy persons (control group) were studied. In all investigated persons activities of alpha 1-proteinase inhibitor (alpha 1-PI), alpha 2-macroglobulin (alpha 2-M), alpha 2-antiplasmin (alpha 2-AP), antithrombin III (ATIII) and plasminogen activator inhibitor type 1 (PAI-1) were determined. RESULTS: Patients with uc had higher level of alpha 1-PI than control group. In patients with active uc lower alpha 2-M activity was found than in patients with us remission. Alpha 2-M activity below 100% had 58% sensitivity and 71% specificity, and predictive value (odds ratio = 6.25) for histologic appearance of active uc. The correlation between alpha 2-M value and uc activity and time of symptoms duration before the study start was found. Patients with inflammatory process beyond sigmoid had lower alpha 2-AP activity than patients with less extended inflammatory process. CONCLUSIONS: Inflammatory process in large bowel changed the level of plasma serine protease inhibitors activities, what was a result of acute phase reaction (alpha 1-PI) and inhibitors consumption in inflammatory processes (alpha 2-M, alpha 2-AP). PMID- 11475846 TI - [Co-existence of reduced femur neck density and reduced skin thickness]. AB - OBJECTIVE: In peri- and postmenopausal women, the association of skin thickness with bone mass is well described, and a low skin thickness is a useful predictor of osteoporosis. In this study the association between skin thickness and bone mass of femoral neck in peri- and postmenopausal women was assessed; and the potential for skin thickness as a screening test for osteoporosis evaluated. DESIGN/METHODS: Skin thickness was measured at the arm by ultrasonography (probe 22 MHz). Bone mass was measured at the neck of femur by dual-energy X-ray absorptiometry. One hundred and three peri- and postmenopausal women were studied. RESULT: Successive decrease of skin thickness going along with the loss of femoral neck bone mass was found. There was a statistically significant difference in skin thickness between group of females with normal bone mass and group with osteopenia (p < 0.05) and osteoporosis (p < 0.01). CONCLUSIONS: Ultrasound skin measurement is a very simple method which can be used in the diagnostics of osteopenia and osteoporosis of the neck of femur in peri- and postmenopausal women. PMID- 11475847 TI - [Use of stents in nephrology]. AB - Stenting is a modern and progressively developing methods in the treatment of blood vessel stenosis of different localization. Currently it is applied in all medical disciplines. The following article presents general characteristics of stents, methods of their implantation, current therapeutic indications for this procedure and treatment results of renal artery, arterio-venous fistulae and ureteral stenoses. PMID- 11475848 TI - [Glucose homeostasis. II. Homeostasis disturbances--hypoglycemia in newborns and infants]. AB - Since glucose is a vital fuel for the brain, hypoglycaemia demands prompt recognition, effective treatment, and accurate identification of the cause. However, pointing the cause, especially in children, can be and often is difficult. Neonatal hypoglycaemia is of a particular importance for paediatricians, since it is, when prolonged or recurrent, a preventable cause of mental retardation and permanent neurological damage. Neonatal hypoglycaemia can be transient or persistent and both of these groups can be further subdivided into two subgroups, where the aetiology of hypoglycaemia is primarily due to decreased hepatic glucose production or to increased peripheral glucose utilization. This article describes first part of causes of neonatal hypoglycaemia. PMID- 11475849 TI - [Therapeutic use of potassium citrate]. AB - Therapeutic indications of potassium citrate include: 1. Oxaluric renal stone disease and some cases of uric acid stone disease. Prevention of stone formation in patients with renal polycystic disease. Prevention of stone relapse after ESWL or lithotomy; 2. Distal renal tubular acidosis complicated by hypercalciuria, mainly in children. 3. Renal hypercalciuria and hyperoxaluria. 4. Prevention of renal complications at the time of glaucoma treatment with acetazolamide. 5. Potassium supplementation during treatment of hypertension. Potassium citrate is usually contraindicated in the case of: 1. Urinary tract infection. 2. Struvite renal stone disease. 3. Hyperpotassemia and advanced chronic renal failure. 4. Peptic ulcer or gastritis. 5. Gastrointestinal bleeding. 6. Disorders of coagulation, crural varices. 7. Metabolic alkalosis. Potassium citrate, when used at therapeutic doses, is to be considered as quite safe. The average daily dose even if admitted as a single dose day engages 60-75% of free renal capacity for potassium excretion. Physiologic and therapeutic citrate concentration in urine exceeds much those available for other inhibitors. The therapeutic dose does not induce any significant changes in any biochemical or endocrine parameter of blood except mild transient metabolic alkalosis. The decrease of urine calcium and increase in oxalate calcium phosphate excretion is observed. In hypo-cytriaturic patients the response to therapeutic dose of citrate is smaller. One-year remission of stone disease is observed in 70-75% cases. PMID- 11475850 TI - [Child abuse as a problem of the family physician]. AB - There are four major forms of child abuse: physical abuse, emotional abuse, sexual abuse and neglect. Various combination of these four aspects often exists in the individual child and clinical manifestation can not be so clear-cut. In this paper the authors present some problems of child abuse syndrome (taking into consideration risk factors, clinical symptoms, recognition and methods of treatment) whose knowledge may be helpful for the family doctor who comes into contact with such children. PMID- 11475851 TI - [Current diagnosis of gastroduodenal reflux and biliary gastritis]. AB - Though primary and secondary duodenogastric reflux (DGR) have been accepted in the medical literature as separate clinical units, reliable and standardised methods of detection have not been known since long. Therefore, the role of DGR in the pathogenesis of upper GI tract diseases makes a problem. So far applied measurement techniques allow only indirect diagnosis of DGR, which is often unphysiological and not objective. These methods do not allow also quantitative evaluation. Unsatisfying results obtained with the use of these methods and further search for the effective system of 24-hour monitoring of bilirubin concentration, which is indirect marker of alkaline reflux Bilitec 2000 is the most physiologic technique of ambulatory recording of the retrograde duodenogastric reflux. It makes possible objective diagnosing of alkaline gastritis as a result of excessive exposure of gastric mucose to destructive effect of bile and pancreatic juice. PMID- 11475852 TI - [Ductal calculi as a diagnostic and therapeutic problem in patients qualifying for laparoscopic cholecystectomy]. AB - Currently used techniques of CBD lithiasis treatment in candidates for laparoscopic cholecystectomy were presented. Advantages and drawbacks of nowadays most popular sequential approach (ERCP + laparoscopic cholecystectomy) were discussed. Technique of laparoscopic CBD exploration, indications, pros and cons were presented. PMID- 11475853 TI - [Use of laparoscopic technic for treatment of patients with reflux diseases]. AB - Pathophysiology, symptomatology and diagnostic work-out in gastroesophageal reflux disease was presented. Treatment strategies and indication for surgery were discussed. Detailed description of the laparoscopic Nissen fundoplication was given. Complications, drawbacks and advantages of this procedure were discussed. PMID- 11475854 TI - [The effect of changes in stroke volume on QT dispersion during long-term DDD and VVI pacing]. AB - The aim of this study was to evaluate the alterations of stroke volume (SV) on the QT dispersion (QTD) as a result of different pacing modes (DDD, VVI) and programmed AV delays (AVD). MATERIAL AND METHODS: We studied 60 patients (pts) (22 F, 38 M) in mean age 67.5 +/- 7.5 yrs in whom DDD pacemakers were implanted due to complete atrioventricular block (AVB III degree). 10 pts had coronary artery disease (CAD)--"CAD" subgroup, 14 pts arterial hypertension (AH)--"AH" subgroup, 27 pts CAD and AH--"AH + CAD" subgroup. The control group consisted of 9 pts without structural heart disease, but with AVB III degree, paced in DDD mode. Previous myocardial infarction was recognised in 7 patients in "CAD" subgroup and in 7 patients in "AH + CAD" subgroup. Left ventricular hypertrophy was detected in 4 men in "AH" subgroup and in 8 patients in "AH + CAD" subgroup. In all cases basic rate of the pacemaker was programmed at 70/min. Resting ECG showed all atrial and ventricular complexes captured. AVD optimization was based on the measurements of SV by Doppler echocardiography. QT intervals (QT) were measured from 12-lead ECG at 50 mm/sec. speed. QTD was calculated as the difference between maximal and minimal QT. It was measured at optimal AVD (with highest SV--opt. DDD) and "unoptimal" AVD (unopt. DDD) programmed AV intervals and then in VVI mode (with lowest SV) after 24 hrs following reprogramming of the pacemaker. RESULTS: In the whole population, in DDD mode, we found a strong correlation between SV and QTD (r = -0.655; p < 0.01). In VVI mode SV was significantly lower than in unopt. DDD (78.7 ml vs 90.0 ml, p < 0.001) and QTD was significantly greater (74.3 ms vs 69.2 ms, p < 0.05). The modifications in AVD and conversion of pacing mode (VVI to DDD) leading to maximal SV resulted in decreased QT dispersion. The highest SV (128.9 ml at opt. DDD) and the lowest QTD (48.3 ms at opt. DDD) were seen in the controls, but changes in dispersion were the least and insignificant during different pacing modes. In CAD; AH + CAD pts, we observed the lowest SV (61.7 ml; 74.9 ml at VVI respectively) and the highest QTD (77.8 ms; 80.9 ms at VVI respectively). In these pts, the modifications of AVD and conversion of pacing mode (VVI to opt. DDD) leading to maximal SV (106.8 ml; 115.9 ml in CAD and AH + CAD pts, respectively) resulted in decreased QTD (53.9 ms; 56.6 ms, respectively). During DDD pacing there was significant correlation of QTD to SV (r = -0.680; r = -0.656, respectively). In CAD pts with myocardial infarction or in AH pts with LV hypertrophy the correlation of SV to QTD was stronger (r = -0.937, r = -0.886, p < 0.025, respectively). We didn't find the correlation in the control group (r = -0.318). CONCLUSIONS: Programming of different AVD and pacing modes significantly influences QTD in CAD and/or AH pts. QTD reflects hemodynamic status of these paced pts. PMID- 11475855 TI - [Changes in QT dispersion during the exercise test in women with syndrome X]. AB - The aim of the present study was to examine the effect of exercise test on QT dispersion (dQT) and to compare the result in women with syndrome X with women with coronary artery disease and normal subjects. We examined 53 women in mean age 54.2 +/- 9.2 who were divided into groups: 20 women with one-vessel coronary artery disease (group I), 19 women with syndrome X (group II) and 14 healthy control women (group III). All subjects underwent a modified Bruce protocol exercise test and QT intervals were measured manually at rest and peak exercise. The value of dQT was calculated as a difference between the longest and the shortest measured value in each of the 12 ECG leads. Corrected QT (dQTc) dispersion was measured after the QT interval was corrected with Bazett's formula. There were no significant differences in rest values of dQT between groups but rest dQTc was significantly greater in group I and II then in group III. We observed significant increase in dQT and dQTc on peak exercise in group I and II when compared with group III. CONCLUSION: At rest dQTc is significantly greater in women with syndrome X and coronary artery diseases in comparison with control subjects. The exercise caused increase in the value of dQT and dQTc both in women with syndrome X and coronary artery disease. The value of dQT and dQTc makes impossible to differentiate between women with syndrome X and women with coronary artery disease. PMID- 11475856 TI - [Evaluation of lipid peroxidation and acid-base status in cord blood of newborns after diabetes in pregnancy]. AB - The purpose of our study was to evaluate lipid peroxidation products and scavenging enzyme activity in placenta and cord blood as well as the estimation of acid-base status and blood gases. Seventy five pregnant patients and their newborns were investigated. Twenty eight had pre-gestational diabetes mellitus (PGDM) and 19 gestational diabetes mellitus (GDM). The following parameters were measured: malondialdehyde (MDA) concentrations, glutathione (GSH) levels, the activity of CuZn dismutase (SOD) (Bioxytech, France). Base excess, pO2, pCO2 and pH were measured in arterial and venous samples. Statistical analysis was performed using Mann-Whitney U test. MDA levels and GSH content increased significantly, while SOD activities declined in diabetic group. Newborns of PDGM mothers had essentially diminished pH and rised both, pCO2 and base deficit. There were no any significant differences in parameters of acid-base balance in newborns of patients with GDM as compared with healthy patients. Our results suggest, that in diabetic patients the fetuses are exposed to increased oxidative stress. The evaluation of antioxidant defence and lipid peroxidation, apart from routine measurement of acid-base balance, might serve as a useful marker of fetal distress in diabetic patients. PMID- 11475857 TI - [Hypertension in patients with diabetes mellitus--selected pathogenetic and therapeutic aspects]. AB - The pathogenesis of hypertension in diabetes type 1 and type 2 is different. Diabetic nephropathy is regarded as the most essential factor contributing to the development of hypertension in patients with diabetes mellitus type 1. Obesity, insulin resistance and hyperinsulinaemia are responsible for hypertension in diabetes mellitus type 2. In both types of diabetes, hypertension is involved in fast progress of diabetic renal disease. Antihypertensive treatment in diabetic patients should include: non-pharmacological interventions, drug-therapy, regular blood pressure monitoring, educational efforts. ACE-inhibitors, calcium antagonists, diuretics, beta-blockers, angiotensin II receptor antagonists and alpha 1-blockers are used as antihypertensive agents in diabetic patients. Neutral endopeptidase inhibitors are the new, promising therapeutic option. PMID- 11475858 TI - [Silent myocardial ischemia]. AB - Nowadays the cardiovascular diseases particularly ischaemic heart disease (IHD) are the most frequent causes of death in Poland. Some of patients with IHD are completely asymptomatic. These subjects are more susceptible to sudden coronary events due to lack of diagnosis and treatment. Cohn divided patients with asymptomatic ischaemia (AI) into three groups: completely asymptomatic, asymptomatic patients after myocardial infarction, patients with painful angina who have some ischaemic events asymptomatic. Some causes of AI are: increased pain threshold, increased beta-endorphin levels, impairment of pain pathways, smaller ischaemic regions in comparison with painful angina, psychological factors, transient platelet microaggregates. Estimated prevalence of AI is about 2-4% of total population and is larger in the group of patients with multiple coronary disease risk factors especially with diabetes mellitus (autonomic neuropathy). In the patients after myocardial infarction the prevalence of AI is between 30-70% and it is associated with poorer prognosis. In subjects with painful angina 70-80% of total ischaemic episodes detected by 24-hour ECG monitoring is asymptomatic. The most useful methods for diagnosis of AI are ECG exercise test and ambulatory 24-hour ECG monitoring, although they may sometimes produce false positive results. Other tests are not widely performed and their use is restricted to specific circumstances. Some cases are finally solved by coronary angiography. Although screening in whole population is not cost effective, but in some groups is necessary (people with many risk factors of IHD, people of certain professions--plane pilots, etc.). Treatment of AI does not vary from treatment of symptomatic IHD. Basic drugs used are: aspirin, beta-blockers, calcium channel blockers, long time acting nitrates. Positive effect of statins is also observed. The most beneficial is invasive treatment--CABG is more efficient than PTCA. Moreover the treatment of symptomatic IHD should be oriented not only to eliminate the symptoms but also to withdraw episodes of silent ischaemia confirmed by 24-h ECG monitoring or ECG exercise test. PMID- 11475859 TI - [Atrial septal defect in adults--opinions, procedures and controversies]. AB - Atrial septal defect (ASD) accounts for 30-40% of congenital heart disease in adults diagnosed after the age of 40 and is after bicuspid aortic valve and mitral-valve prolaps the most common congenital cardiac malformation in adults. We have discussed current views on the pathological role of ASD in adults and controversies regarding its treatment. It is expected that increasing understanding of ASD pathophysiology, improved diagnostic methods and the possibility of transcatheter closure of interatrial defects will improve the treatment of patients with ASD. PMID- 11475860 TI - [TAFI--thrombin activated fibrinolysis inhibitor ]. AB - This is a review of literature data concerning Thrombin Activatable Fibrinolysis Inhibitor (TAFI) and its role in haemostatic system in vitro and in vivo. This is the glycoprotein, which was identified recently by three independent groups of researchers. TAFI is converted to its active form by thrombin-thrombomodulin complex. TAFIa removes lysine residues from fibrin net, making impossible formation of plasminogen, t-PA and fibrin complex. It causes impairment of plasmin generation, and by this way suppression of fibrinolysis. PMID- 11475861 TI - [Autoimmune hepatitis: pathogenesis, clinical features and treatment]. AB - Autoimmune hepatitis (AIH) is a chronic liver disease of unknown etiology. The incidence of AIH keeps rising, most probably because of increasing availability of the diagnostic tools permitting thorough differential diagnosis amongst liver diseases presenting histologically as chronic active hepatitis. In this review we focus on clinical aspects of autoimmune hepatitis and discuss pathogenesis, clinical features and treatment of AIH. PMID- 11475862 TI - [Current principles for diagnosis and treatment of hydatid disease]. AB - Hydatid disease is a parasitic disease caused by one of two species of Echinococcus: Echinococcus granulosus (cystic hydatid disease) or Echinococcus multilocularis (alveolar hydatid disease). Widely accepted criteria of diagnosis of hydatid disease are based on the result of ultrasonography suggesting the presence of hydatid cyst and positive results of serologic tests. The serologic tests should begin with a screening test (ELISA or haemagglutination), and then should be verified using a western-blot method in order to detect the reaction with 8 kDa fraction of Echinococcus antigen. Further differentiation of the infecting species may be accomplished with the aid of Em2 plus or Em-18 ELISA tests which detect antibodies specific for E. multilocularis. The final diagnosis is based on the histologic examination of hydatid lesion or detected protoscolex of tapeworm in cystic liquid. Treatment of cystic hydatid disease consist of surgical therapy, percutaneous drainage (PAIR), chemotherapy and long-term observation. Treatment of alveolar hydatid disease is based on radical surgical resection of parasitic lesion and many years of chemotherapy. In case of chronic liver failure the transplantation is necessary. PMID- 11475863 TI - [Progress in tests for cavum septi pellucidi in children and adults]. AB - Persistent cavum septi pellucidi (CSP) is a congenital anomaly of the midline, regarded as a marker of cerebral dysgenesis. It is more commonly encountered in patients with disturbances of intellectual development. Experimental data point to the role of CSP in memory and learning-related processes, and also to its association with disturbances of bioelectric activity of the brain. Various authors emphasize its occurrence in schizophrenia, various motor syndromes and some selective developmental deficits. Of a great significance in the better understanding of CSP will be the assessment of the bilateral brain bioelectric activity in children, as well as neuropsychological evaluation of the very patients at subsequent developmental stages. This will allow for a detailed assessment of their quality of life and neurodevelopmental problems, as well as for an early introduction of thus identified therapeutic management. PMID- 11475864 TI - [Idiopathic headaches in children and adolescents]. AB - Author presents new epidemiological data in prevalence of headaches in children and adolescents. She precises tension headaches and pathogenesis of migraine and reminds difficulties in its differentiation. She points out to endothelin-1 importance in this process. PMID- 11475865 TI - [Laparoscopic surgery of the spleen]. AB - Indications for laparoscopic a surgery of the spleen were presented. Detailed description of the laparoscopic splenectomy and operation of the splenic cyst were given. Complications, drawbacks and advantages of this procedure were discussed. PMID- 11475866 TI - [Recurrent vasovagal syncope and indications for pacemaker implantation--report of two cases]. AB - Recurrent syncope is a common clinical problem and a significant diagnosis and therapeutic challenge. Head-up tilt testing is commonly used in the evaluation of patients with vasovagal syncope. Analysis of the type of reaction leading to event is a base to the therapeutic method. Documented symptomatic bradycardia or asystole during head-up tilt testing is an indication to dual-chamber pacemaker implantation. PMID- 11475867 TI - [Thyroid crisis diagnosed on the basis of anatomic-pathologic examination]. AB - Graves-Basedow disease is the most common cause of hyperthyroidism. Rhabdomyolysis is occasionally associated with metabolic coma or severe electrolyte disturbances. We describe rhabdomyolysis accompany ing thyroid crisis. A 48-years old man admitted with hypovolemic shock developed rhabdomyolysis, congestive heart failure, hepatic failure and renal failure and then died. It is interesting that correct diagnosis was done on the basis of microscopic examination of specimens of the thyroid gland. PMID- 11475868 TI - [The all-in-one technique]. PMID- 11475869 TI - [The phenomenon of "nanoleakage" in composite-dentin transition by the use of adhesives]. PMID- 11475870 TI - Assessment of elimination of neonatal tetanus, Zimbabwe. PMID- 11475871 TI - Onchocerciasis (river blindness). PMID- 11475872 TI - Meningococcal disease, serogroup W135 (update). PMID- 11475873 TI - Dracunculiasis, Kenya. PMID- 11475874 TI - Development and implementation of a Hope Intervention Program. AB - PURPOSE/OBJECTIVES: To describe the development and evaluation of the Hope Intervention Program (HIP), designed to enhance hope, based on the Hope Process Framework. DESIGN: Descriptive and evaluative. SETTING: Outpatient oncology clinics of two large hospitals. SAMPLE: Convenience sample--38 adults with first recurrence of cancer. METHODS: Questionnaire completed at end of last HIP session, three months, six months, and nine months. MAIN RESEARCH VARIABLES: Helpfulness of the HIP components in maintaining hope. FINDINGS: Participants used intervention strategies that represented all four attributes of hope, with specific strategy use varying across time and individuals. CONCLUSIONS: Data suggested that HIP positively affected the participants' rebuilding and maintenance of hope. IMPLICATIONS FOR NURSING PRACTICE: HIP has the potential to positively influence hope, and it supports the strategic vital role that nurses play in implementing research-based strategies designed to engender hope in their patients and prevent hopelessness. PMID- 11475875 TI - The child's worries about the mother's breast cancer: sources of distress in school-age children. AB - PURPOSE/OBJECTIVES: To describe children's worries when their mothers are newly diagnosed with early-stage breast cancer. DESIGN: Descriptive, qualitative study. SETTING: Private family homes. SAMPLE: Case intensive interviews with 16 children who ranged in age from 11-18 years at the time that interviews were conducted and who had been 8-12 years of age when their mothers were diagnosed with early-stage breast cancer. METHODS: Semistructured interviews with the children were audiorecorded, transcribed, and inductively coded into categories of distinct worries about their mothers' breast cancer. MAIN RESEARCH VARIABLES: Children's descriptions of their worries and confusion resulting from their mothers' breast cancer diagnoses. FINDINGS: The children voiced nine categories of worry during the interviews: worrying that the mother was going to die; feeling confused; worrying that something bad would happen; worrying about the family and others; worrying when the mother did not look good; worrying that their mothers would change; wondering if the family would have to cut back financially; worrying about talking to others; and wondering if they, the children, would get cancer. CONCLUSIONS: Children of mothers with breast cancer experience multiple worries concerning their mothers, their families, and themselves. The data revealed that they attempted to make sense of their mothers' illness for themselves and imagined how it might affect their own lives in the future. IMPLICATIONS FOR NURSING PRACTICE: Programs and materials need to be developed that help parents address the multiple worries that children whose mothers have early-stage breast cancer experience. PMID- 11475876 TI - Relationship between fatigue and nutritional status in patients receiving radiation therapy to treat lung cancer. AB - PURPOSE/OBJECTIVES: To describe the relationship between fatigue and nutritional status in patients receiving radiation therapy for lung cancer. DESIGN: Prospective, observational study. SETTING: Radiation therapy department of a large mid-western cancer center. SAMPLE: 45 adults with primary cancer of the lung receiving outpatient primary or adjuvant radiation therapy. METHODS: Measurements taken before radiation therapy, at week four of treatment, and at completion included the Piper Fatigue Scale, prealbumin levels, and weights. MAIN RESEARCH VARIABLE: Fatigue, nutritional status. FINDINGS: Weight loss over the course of treatment was significant but did not correlate with fatigue; fatigue did not change significantly during the measurement period. CONCLUSIONS: Fatigue and nutrition are major problems for patients with lung cancer, but nutritional changes do not correlate with fatigue. An increase in fatigue during radiation therapy was not identified. IMPLICATIONS FOR NURSING PRACTICE: Nurses should continue efforts to intervene with the problems of nutrition and fatigue. The study should be repeated using different fatigue Instruments and with other cancer populations. PMID- 11475877 TI - Breast cancer detection: knowledge, attitudes, and behaviors of women from Pennsylvania. AB - PURPOSE/OBJECTIVES: To assess relationships among breast cancer detection behaviors and selected variables in healthy women. DESIGN: Correlational study. SETTING/SAMPLE: A sample of 1,000 women was selected randomly from the 16,500 members of the General Federation of Women's Clubs of Pennsylvania. Respondents (N = 538; 54% response rate) were predominately white, well educated, lived in urban areas, and had an average age of 60 years. METHODS: Mailed packets with a professionally designed, scannable survey instrument that included questions related to detection behaviors, a risk index, health behaviors, attitudes, and knowledge. MAIN RESEARCH VARIABLES: Breast cancer detection behaviors: mammography, clinical breast examination (CBE), breast self-examination (BSE). Structural/demographic variables: age, education, residence, knowledge of breast cancer and detection methods, teaching history, encouragement, and risk index (family/medical history). Predisposing variables: susceptibility, benefits/barriers, confidence, social norms and influence, and general health motivation. FINDINGS: Women reported moderate/high adherence to recommendations for early detection of breast cancer. Mammography behavior was predicted by older age, being encouraged by a doctor or nurse, and greater risk. CBE predictors were greater knowledge and risk along with greater benefits, social norms, and health motivation. BSE behavior was predicted by having had BSE technique checked, greater knowledge, greater risk, decreased barriers to BSE, and higher health motivation. CONCLUSIONS: Common predictors of breast screening behaviors include risk (family/medical history), knowledge, and general health motivation. IMPLICATIONS FOR NURSING PRACTICE: Educational efforts can be designed specifically to influence variables related to compliance with early breast cancer detection behaviors. PMID- 11475878 TI - Physical activity and fatigue during radiation therapy: a pilot study using actigraph monitors. AB - PURPOSE/OBJECTIVES: To describe the relationship of patterns of physical activity and fatigue during radiation therapy. DESIGN: Prospective, descriptive, repeated measures, pilot study. SETTING: Outpatient radiation oncology clinic at a large medical center. SAMPLE: Seven adult subjects with cancer who received a six-week course of external beam radiation therapy to the trunk (including breast, chest, or abdomen). METHODS: Wrist actigraphs were used to measure physical activity for 72 hours during the second and fifth weeks of therapy. The Fatigue-Inertia and Vigor-Activity subscales of the Profile of Mood States (POMS) and the Linear Analogue Scale-Fatigue (LAS-F) were used to measure perceptions of fatigue at the beginning and the end of the week of the two collection periods. The Symptom Distress Scale (SDS) assessed symptom severity. Subjects recorded their physical activities in a five-day diary. MAIN RESEARCH VARIABLES: Physical activity, fatigue, and six-week course of external beam radiation therapy. FINDINGS: Physical activity levels were highest at the end of the week at both week two and week five. As measured by POMS and LAS-F, perception of fatigue decreased at the end of the week. SDS showed minimal change in symptom distress. CONCLUSIONS: Contrary to expectations, activity increased during treatment and fatigue decreased. This agrees with current work supporting the benefits of exercise during cancer treatment. IMPLICATIONS FOR NURSING PRACTICE: Physical activity during treatment, as compared to inactivity, may help to reduce fatigue. PMID- 11475879 TI - On the frontlines of old battles. PMID- 11475880 TI - More research is needed on the importance of certification. PMID- 11475881 TI - Communication skills: breaking bad news in the clinical setting. AB - Oncology nurses need to develop expert communication skills, especially when delivering bad news to patients and families. Patients and families differ in their needs for levels of information, interpretation of information delivered to them, and responses to unfavorable news. When bad news is delivered in a sensitive and caring manner, morale can be maintained, and the process of coming to terms with illness or death is possible through realistic expectations and hope. It is valuable for nurses to elicit patients' or family members' understanding of what is happening to them, and what they need is key in the process of communicating in a crisis time. Also, skill in responding to feelings and not the content of expression is necessary to the process of recovery. So many improved cancer treatments exist, but the goal of effectively communicating bad news with respect and caring is just as important as treating the person who happens to have cancer. PMID- 11475882 TI - Mastering the intricate maze of metastasis. AB - The metastatic process is an intricate maze of cellular malfunction and deregulation. Many complex steps in the process are necessary for cancer cells to develop a metastatic site. Awareness of these steps has increased oncology nurses' understanding of the biologic process and furthered hopes for interventions to prevent metastasis. Despite these advances, [table: see text] metastatic disease is the leading cause of cancer morbidity and mortality. Nurses who master the concepts of metastasis will appreciate the impact of this phenomena on patients' quality of life, be better prepared to provide information on new treatment strategies to patients and the public, and be able to provide patients with hope for the future. PMID- 11475883 TI - I'm coping as fast as I can: psychosocial adjustment to cancer and cancer treatment. PMID- 11475884 TI - Behavioral oncology research: a new millennium. PMID- 11475885 TI - Mastery of postprostatectomy incontinence and impotence: his work, her work, our work. AB - PURPOSE/OBJECTIVES: To describe couples' experiences of postprostatectomy incontinence and impotence. DESIGN: Descriptive, qualitative. SETTING: Northeastern U.S. metropolitan area. SAMPLE: Subsample of 20 (10 control and 10 intervention) couples from a large quantitative clinical trial of a Standardized Nursing Intervention Protocol (SNIP) postprostatectomy. METHODS: Interviews were conducted using a semistructured guide. Data were analyzed using grounded theory techniques. MAIN RESEARCH VARIABLE: Couples' experiences of coping with postprostatectomy incontinence and impotence. FINDINGS: Managing postprostatectomy incontinence and impotence required work. Men's work focused on regaining mastery and encompassed understanding incontinence as healing, mastering incontinence, networking, confronting impotence and putting it into perspective, and prioritizing. Wives were supportive by managing anxiety, encouraging mastery, putting impotence into perspective, and reassuring their spouses. Established routines brought couples through the experience together while strengthening intimacy. SNIP couples found the nurses to be sources of information, support, and affirmation. CONCLUSIONS: Couples worked to deal with postprostatectomy incontinence and impotence within the context of surviving cancer and maintaining a loving relationship. This gave unique meaning to their symptoms and led the couples to value the fact that the men were alive and work toward regaining mastery. Mastery emerged as a key concept from the findings. IMPLICATIONS FOR NURSING PRACTICE: Nurses can gain from an enhanced understanding of postprostatectomy incontinence and impotence as meaningful within the greater context of patients having had cancer. Nurses can hasten couples' abilities to regain a sense of mastery by providing information, supporting couples' work, providing positive affirmation, and being available. PMID- 11475886 TI - Sentinel lymph node biopsy in breast cancer: scientific rationale and patient care. AB - PURPOSE/OBJECTIVES: To describe the history of sentinel lymph node biopsy (SLNB), review the current scientific literature comparing the benefits and shortcomings of SLNB with traditional axillary lymph node dissection, and describe the nursing role that should be employed when instructing patients who are considering SLNB. DATA SOURCES: Journal articles, published research data, and clinical experience. DATA SYNTHESIS: Traditional axillary lymph node dissection has the potential to cause serious complications such as lymphedema, scarring, numbness, pain, and psychological distress. Given that approximately 70% of women with early-stage breast cancer will have no evidence of regional lymph node involvement at the time of surgery, determining who is likely to have negative nodes will spare women these potential complications. SLNB can significantly minimize the morbidity associated with axillary lymph node dissection while providing accurate diagnostic and prognostic information. CONCLUSION: SLNB has been well documented in the scientific literature from multiple phase III clinical trials as an accurate, safe, and fiscally conservative alternative to traditional axillary lymph node dissection for women who present with early-stage breast cancer. Furthermore, future results from multicenter, randomized clinical trials now under way ultimately will determine the role for SLNB in the years to come. IMPLICATIONS FOR NURSING PRACTICE: Nurses in the outpatient setting can help to minimize the anxiety and fear that patients have when they are considering SLNB versus the more traditional axillary lymph node dissection. Oncology nurses also serve as resources to other nurses, healthcare professionals, and the public as more information is learned concerning the role of SLNB in early-stage breast cancer. PMID- 11475887 TI - [Advances in the development of new antitubercular agents from a group of ortho condensed heterocyclic compounds. Part 1. Compounds containing one heteroatom per six-membered ring]. AB - Tuberculosis and other mycobacterial diseases are considered to be one of the most important problems of contemporary health service. Since 1985 and particularly in the 1990s and at present the search for new structures of antimycobacterial agents have ranked among the foremost areas of chemotherapeutic research. The present review paper is already the 16th communication in a group of review papers about the development of new antituberculotics in recent 15 years. The literature search is based on the journal Chemical Abstracts, Current Awareness in Biomedicine, part Mycobacteria, and original papers. Current Awareness in Biomedicine is, however, a very imperfect literature search source, recording only a fraction of communications. Though none of the ortho-condensed heterocyclic compounds has been introduced into practice, the in vitro evaluated activities of some of them equalled those of routine antituberculotics. Polycyclic systems make it possible to widely model the structures, and therefore the present review paper can inspire a search for new structures of antimycobacterial agents. PMID- 11475888 TI - [Enantiomeric separation of drugs based on macrocyclic antibiotics]. AB - The present review paper describes the properties of the macrocyclic antibiotics vancomycin, teikoplanin, and ristocetin A, which can be used as stationary phases for separation of the enantiomers of drugs containing a stereogenic centre. It analyzes the results obtained in the separation of amino acids, its derivatives, loxiglumide, clenbuterol, practolol, carnitine, levodopa, methyldopa, ceftazidine, alkoxyphenylcarbamic acid derivatives, dichloroprop, ketoprofen, warfarin, and ergot alkaloids by means of high-performance liquid chromatography (HPLC). PMID- 11475889 TI - [Phytotherapeutic aspects of diseases of the circulatory system. 7. Schisandra chinensis (Turcz.) Baill.): its composition and biological activity]. AB - Schisandra chinensis (TURCZ.) BAILL., originally a Japanese-Manchurian endemite, yields a vegetable drug (Schisandrae fructus) with a number of very utilizable therapeutic effects. The paper reports the results of phytochemical and pharmacological-toxicological studies approximately from the year 1990 carried out both with the drug and, in particular, the principal isolated lignans of the dibenzo[a,c]cyclooctadiene type. The results confirm the validity of the historical use of the drug, in particular as a hepatoprotective, adaptogenic, and antioxidative agent. It is obvious that a very positive therapeutic effect based on the use of a complex mixture of its principal constituents because their biological effects are complementary and potentiate each other. At the same time, some lignans (e.g. gomisin A, gomisin N) are interesting as new prospective medicines. PMID- 11475890 TI - [Unused drugs returned to pharmacies. I]. AB - Drugs that have not been administered and have been stockpiled in households represent a medical, professional, and ecological problem. The present paper analyses and evaluates these drugs from qualitative, quantitative, and financial aspects. The material under study consisted of drugs returned to one pharmacy within a period of 1.5 months (April-May) in the year 1999. Out of set I (1,091 packages), 721 packages (66.1%) were used and 370 (33.9%) packages were original, i.e. unused. The expiration period was over in 78.3%, 16.8% could still be used, and in 4.9% it was impossible to determine their expiration period. The value of the returned drugs was 76,000 Czech crowns. PMID- 11475891 TI - [Pharmacotherapy of osteoporosis in ambulatory practice and its pharmaceutical aspects]. AB - Within the framework of studies of pharmacotherapy of the diseases of the muscular and skeletal system, the paper aimed to analyze the pharmacotherapy of osteoporosis. The significance of the problems in the field of research of epidemiology of osteoporosis requires the use of all available methods. With regard to the fact that the epidemiological situation in osteoporosis is also reflected in the consumption of drugs, the paper aimed to contribute to the solution of the problem by means of an analysis of the pharmacotherapy of osteoporosis. The results of the analysis have confirmed the relationship between the prevalence of the disease of osteoporosis and drug consumption. The paper demonstrated an increase in the total consumption of drugs prescribed for osteoporosis in 1998 and 1999 in comparison with 1997 as well as an increase in the consumption according to the age and sex. A statistically significant difference was recorded in the consumption of drugs prescribed for osteoporosis with regard to women. The analysis showed changes in the assortment of prescribed drugs as well as changes in the management of the therapy of osteoporosis. PMID- 11475892 TI - [Effect of losartan on antioxidant status in rats with diabetes mellitus]. AB - The aim of this study was to monitor the effect of the administration of antihypertensive drug losartan on: (1) the antioxidant status of rats with experimental insulin-dependent diabetes mellitus; (2) oxidative damage which is represented by the production of compounds which can react with thiobarbituric acid (TBARP), and (3) some metabolic parameters. Losartan administration did not significantly influence the concentration of glucose, cholesterol, triacylglycerol, and uric acid in the plasma of control and diabetic animals. In the liver tissue, the concentration of triacylglycerols decreased after losartan administration, but the concentration of cholesterol did not change. The present authors have found that losartan administration increased the levels of water solubile antioxidants in the plasma of diabetic rats, which can result in a decrease of the TBARP levels in the plasma of diabetic rats. PMID- 11475893 TI - [Analytic evaluation of BL 443 pivalate]. AB - The paper presents the results of an analytical evaluation and a study of some physicochemical properties of a new potential drug of the prodrug type--the substance BL 443 pivalate with an assumed antidysrhythmic effect. The structure of the substance has been confirmed by elemental analysis, IR and UV spectroscopy. The following parameters were determined: melting point, solubility, dissociation constant, experimental partition coefficient in four different systems, and surface activity. The chromatographic behaviour of the drug on a thin layer (adsorption and partition chromatography) was also investigated. Acidimetric titration in non-aqueous medium and spectrophotometry in the ultraviolet region of the spectrum at the wavelength of the second absorption maximum of the drug was employed to determine the content of the drug in pure substance. PMID- 11475894 TI - [Anti-radical activity of certain anti-ulcer agents and local anesthetics]. AB - The free radical scavenging effect of the hydrochloride of (+/-)-trans-2-(1 pyrrolidinyl-)cyclohexylester of 3-(n)-pentyloxy carbanilic acid, pentacaine (trapencaine I.N.N), and its derivatives and other substances, lidocaine, stobadine, mannitol, and ascorbic acid was tested in the presence of diphenyl-p picrylhydrazyl (decrease in absorbance). In this method, more pronounced antiradical activity was observed with trapencaine, more effective than stobadine, ascorbate, and mannitol. Trapencaine derivatives and lidocaine were less effective in comparison with trapencaine. After introducing a methylene group into the hydrophilic moiety of the trapencaine molecule (stereoisomers P 18, P-20), there was a loss of antiradical activity, but the cis-isomer is more effective than the transisomer. The results demonstrate the existence of relationships between the chemical structure, antiradical and gastric cytoprotective activity. PMID- 11475895 TI - [Positional clonage and characterization of the bovine myostatin gene]. AB - The double-muscled condition has been intensively selected for in the Belgian Blue cattle breed, where segregation studies have demonstrated the monogenic, autosomal and recessive determinism. This has been confirmed by genetic linkage which located the gene to the centromeric tip of chromosome 2. Our positional cloning strategy, and the discovery of a positional candidate in the mouse, led us to the identification of the causative gene now referred to as the Myostatin gene, since its product downregulates skeletal muscle mass. Disruptive mutations of the gene in cattle have been shown to be responsible for the muscular hypertrophy found in eight european beef breeds. A 15 Kilobases genomic region, including the myostatin gene, has been sequenced and compared in cattle and mice. The murine gene has undergone a complex genetic engineering in order to test different allelic variants in vivo after gene targeting transgenesis. PMID- 11475897 TI - [Paracrine kidney activity and homeostasis: the emergence of nitric oxide. A conceptual perspective]. AB - In nephrology, intensive research has focused in recent years on the interplay between NO produced by the different isoforms of NO synthase (NOS) and complex renal functions. In this regard, the juxtaglomerular apparatus is of particular interest. First, it is the main site for control of renal blood flow by autoregulation and of glomerular filtration rate by tubuloglomerular feedback, as well as of renin secretion. Second, two constitutive NOS, nNOS and eNOS, are expressed, respectively, in the macula densa cells and in the endothelium of the afferent and efferent arteriole. It was thus not unexpected that NO could interact with the physiological variables. Indeed, NO attenuates rapidly the autoregulatory efficacy of renal blood flow as well as the sensitivity of the tubuloglomerular feedback by inhibiting Ca++ influx mainly in the afferent arteriole. On the other hand, renin secretion may be stimulated as well as inhibited by NO. These opposing effects, although as yet unexplained, could be related to the source of NO. If so, the hypothetical dual effect of NO could be secondary to a difference in modulation of each NOS isoform by their specific stimuli and results in secretion or not of renin. This hypothesis seems to be applicable to changes in extracellular volume. PMID- 11475898 TI - [From muscle to action--Hermann von Helmholtz (1821-1894)]. AB - Helmholtz is one of the most brilliant physiologists of the 19th century. His experimental research was respectful for strict empiricism and physical laws, but he admitted that a physiological theory of perception was only possible in the frame of a psychological approach, irreductible to physiology. He is considered as a precursor of the Neurosciences. PMID- 11475899 TI - [From bioethics to the new technology ethics]. AB - Just as what we can all "technoscience" is emerging in our everyday life, a reflection should be conducted concerning the implications of the scientific and technical progress within our society from now on globalised. We will tackle successively: 1. The ambiguities and paradoxes related to the development of new technologies: in the field of bioethics: artificial reproduction, mammal cloning, genetically modified organisms. towards the ethics of new technologies: ethics of information and communication technologies and ethics of space policy; 2. Nature, foundation and characteristics of the ethical approach; the precaution principle must be completed with two other principles: the principle of experience and the principle of vigilance; 3. The modalities of a democratic management of the ethical approach: it is a matter of defining the role of the three main actors, i.e.: experts, politicians and citizens representing public opinion. It is necessary to promote the ethical approach within a democratic context, that is to ensure a dialogue between experts, policy decision-makers and public opinion on all of the applications of science and technology. It is from such a permanent and renewed dialogue that will emerge the image we give from ourselves in the present world. PMID- 11475900 TI - The effect of salinomycin on health status and performance of sows and their litters: a dose titration study. AB - Two hundred and fifty gilts and sows (Dalland parent stock) were divided randomly and allocated into the following five experimental groups: SAL0 = negative controls, SAL20 = 20 mg salinomycin per kg of feed, SAL40 = 40 mg salinomycin per kg of feed, SAL60 = 60 mg salinomycin per kg of feed and SAL80 = 80 mg salinomycin per kg of feed. Each gilt and sow was allocated to one of the five groups after the confirmation of the pregnancy using the ultrasonic method and remained under treatment for two consecutive breeding cycles. Throughout the experimental period several parameters related to sow health status, performance and fertility, as well as to health status and performance of their litters were recorded and calculated. Results indicated that salinomycin improves most of the parameters examined, as it leads to significantly higher (P < 0.05): (i) sow body weight gain during gestation; (ii) number of piglets born alive and weaned; (iii) piglet body weight at birth and at weaning, as well as to significantly lower (P < 0.05): (i) prevalence of thin sow syndrome at weaning; (ii) sow body weight loss during lactation; (iii) weaning-to-oestrus interval; (iv) piglet diarrhoea score during lactation; and (v) preweaning mortality. The best results were obtained at the inclusion levels of 40 and 60 mg salinomycin per kg of feed. PMID- 11475901 TI - Effects of sevoflurane, isoflurane and halotane anaesthesia on fluorescein angiographic phases of dogs: a comparative study. AB - A fluorescein angiography method was developed to compare the onset and the total duration of the fluorangiographic phases between three anaesthetic protocols in six healthy mixed-breed dogs. The animals were anaesthetized three times. Each dog received, as pre-anaesthetic protocol, atropine (10 micrograms/kg intramuscularly), and as a sedative, romifidine (80 micrograms/kg intravenously). Fifteen minutes later, induction of anaesthesia was delivered with propofol (1 mg/kg intravenously) and maintained either with sevoflurane (SEVO group), isoflurane (ISO group) or halothane (HAL group) for 30 min in all cases. Some angiographic, cardiovascular and respiratory variables were registered during the procedure. Recovery times were also registered. Angiographic variables recorded were: onset of the arterial phase (TA), onset of the arteriovenous phase (TAV), onset of the venous phase (TV), complete arterial phase duration (I1), complete arteriovenous phase duration (I2) and I1 plus I2 (I3). Mean heart rate, mean arterial pressure, systolic arterial pressure, diastolic arterial pressure, respiratory rate, tidal volume, arterial oxygen saturation and end-tidal CO2 during SEVO and ISO anaesthesia, were similar in dogs. Minute ventilation and rectal temperature were higher in dogs with SEVO than ISO. HAL produced higher arterial pressures and a lower arterial oxygen saturation than ISO and SEVO. Mean respiratory rate, rectal temperature and minute ventilation were higher in HAL. Pulse rate, end-tidal CO2 and tidal volume were similar in the dogs of the three groups. No differences in recovery times were found. The fluorescein angiographic times were within the normal range. There were no significant differences between protocols in I1, I2 or I3. HAL produced a significant increase of all temporal variables (TA, TAV and TV) when compared with ISO; TA was higher in HAL than SEVO treated dogs. All protocols appear to be safe and effective for inducing and maintaining general anaesthesia in healthy dogs for performing fluorescein angiography. PMID- 11475902 TI - Inter-breed differences in equine forelimb kinematics at the walk. AB - Linear, temporal and angular biokinematic characteristics of the forelimb at the walk in different breeds were determined, highlighting inter-breed differences. Twenty-three healthy stallions were used: ten Andalusians (AN), seven Arabs (AR) and six Anglo-Arabs (AA). Height at the withers was significantly different between groups (P < 0.001). Six trials per horse were recorded using a levelled video camera (sampling frame rate 25 frames/s), digitized and analysed using a semi-automatic movement analysis system. No statistically significant differences in speeds were recorded between breeds (P > 0.05). The only temporal parameter which was similar in the three breeds was the moment at which the hoof reached the highest point in its trajectory. The variables presenting the most significant differences were the percentages of deceleration and propulsion within the stance phase. ANOVA for angular variables showed that the greatest difference was in the range of angular movement of the carpal joint, being higher in AN, due to a lower minimum value. In the fetlock joint, the greatest difference was observed in minimum values, which differed in all three breeds. Significant inter-breed differences were also observed for maximum limb retraction, being lowest in the AN group, followed by the AA and AR groups. This finding was reflected in the angular range of motion, despite smaller differences in the degree of limb protraction; very similar values were reported in all three breeds. As regards the elbow joint, no inter-breed differences were observed in terms of minimum values, whereas differences were recorded for maximum and angular range of motion, higher values being displayed by the AR and AN groups than by AA animals. In conclusion, inter-breed differences may be determined in equine forelimb biokinematics at the walk. This study distinguished between AN, AR and AA horses. PMID- 11475903 TI - Gastro-oesophageal reflux during thiopentone or propofol anaesthesia in the cat. AB - Lower oesophageal pH was monitored in 50 cats anaesthetized with either thiopentone or propofol. Gastro-oesophageal reflux, as evidenced by a decrease in lower oesophageal pH to less than 4.0 or an increase to more than 7.5, occurred in 16% (4/25) and 12% (3/25) of the cats anaesthetized with thiopentone and propofol, respectively, the difference between the two groups being non significant. Reflux usually occurred shortly after the induction of anaesthesia and had a mean duration of about 23 min. The refluxate was always acidic (pH < 4.0). Gastric contents of pH below 2.5 were refluxed on three occasions, two in the thiopentone group and one in the propofol group. Regurgitation and flow of gastric contents from the mouth occurred in only one cat anaesthetized with propofol. None of the cats that exhibited reflux developed any signs of postanaesthetic oesophagitis or stricture formation. PMID- 11475904 TI - The humoral immune response to recombinant nucleocapsid antigen of canine distemper virus in dogs vaccinated with attenuated distemper virus or DNA encoding the nucleocapsid of wild-type virus. AB - This study compared the humoral immune response against the nucleocapsid-(N) protein of canine distemper virus (CDV) of dogs vaccinated with a multivalent vaccine against parvo-, adeno-, and parainfluenza virus and leptospira combined with either the attenuated CDV Onderstepoort strain (n = 15) or an expression plasmid containing the N-gene of CDV (n = 30). The vaccinations were applied intramuscularly three times at 2-week intervals beginning at the age of 6 weeks. None of the pre-immune sera recognized the recombinant N-protein, confirming the lack of maternal antibodies at this age. Immunization with DNA vaccine for CDV resulted in positive serum N-specific IgG response. However, their IgG (and IgA) titres were lower than those of CDV-vaccinated dogs. Likewise, DNA-vaccinated dogs did not show an IgM peak. There was no increase in N-specific serum IgE titres in either group. Serum titres to the other multivalent vaccine components were similar in both groups. PMID- 11475905 TI - Epidural analgesia with a combination of bupivacaine and buprenorphine in rats. AB - We determined the analgesic effects of epidural administration of either bupivacaine at 62.5, 125, 250, and 500 micrograms/kg; buprenorphine at 5 and 10 micrograms/kg; and the combination of bupivacaine at 125 micrograms/kg and buprenorphine at 5 or 10 micrograms/kg, using the tail flick (TF) and colorectal distension (CD) tests in rats and compared the results with those obtained using morphine at 100 micrograms/kg. In both the TF and CD tests, all doses of bupivacaine alone produced potent anti-nociceptive effects, although the effect rapidly diminished after 20-30 min of administration. The administration of buprenorphine at 10 and 5 micrograms/kg produced mild to moderate anti nociceptive effects in both the TF and CD tests, and the effects were relatively constant throughout the 2-h experimental period. Combinations of 5 or 10 micrograms/kg of buprenorphine with 125 mg/kg of bupivacaine produced a significantly higher percentage of maximum possible analgesic effect (%MPE) than that of the calculated additive effect of each drug alone in the TF and CD tests. The analgesic effect of this combination was similar to that of morphine. Minimal ataxia was observed in rats administered this combination. PMID- 11475906 TI - Skeletal muscle fibre characteristics in young and old bulls and metabolic response after a bullfight. AB - Fibre type composition, activities of enzymes such as citrate synthase (CS), 3 hydroxyacyl coenzyme A dehydrogenase (HAD), lactate dehydrogenase (LDH), as well as glycogen, lactate and pH levels were analysed in muscle biopsies (m. gluteus medius) obtained after bullfighting from 10 young and 10 old bulls. No changes were seen in fibre type composition between groups, but the older bulls had higher HAD and LDH activities. Low glycogen concentrations and low pH values were found in both groups, but the lactate concentration after bullfighting was higher in the older group of bulls. The histochemical stain for glycogen revealed that type IIB fibres in both young and old bulls contained more glycogen than seen in type IIA and type I fibres. These results show that young and old bulls have similar muscle fibre type composition, but the metabolic capacity differs, with a higher glycolytic capacity and lactate production in older bulls. Furthermore, it seems that the physical and emotional stress in connection with a bullfight causes a marked depletion of glycogen, especially of type I and IIA fibres. PMID- 11475907 TI - 1-Pyrrolines (3,4-dihydro-2H-pyrroles) as a template for new drugs. PMID- 11475908 TI - Potential tuberculostatic agents: micelle-forming copolymer poly(ethylene glycol) poly(aspartic acid) prodrug with isoniazid. AB - With the objective of obtaining slow-acting isoniazid derivatives, of potential use as chemoprophylactics or chemotherapeutics in tuberculosis, the micelle forming copolymer of poly(ethylene glycol)-poly(aspartic acid) prodrug with isoniazid was synthesized. The derivative obtained was found to be active in Mycobacterium tuberculosis culture, with a minimal inhibitory concentration (MIC) 5.6 times lower than that of the tuberculostatic drug. PMID- 11475909 TI - Inactivation of protein farnesyltransferase by active-site-targeted dicarbonyl compounds. AB - Upon farnesylation by protein farnesyltransferase (FTase), key proteins become compartmentalized in cells. For example, cell membrane localization is essential for the mitogenic role of mutant Ras protein, which acts as a switch for cancer cell proliferation. We report that alpha-dicarbonyl compounds derived from the isoprenoid skeleton or other hydrophobic groups potently obstruct farnesylation of a Ras model peptide by human recombinant FTase in vitro. A geranyl-derived isoprenoid diketone, 5,9-dimethyl-8-decene-2,3-dione, at 17 microM caused a 62% reduction in FTase activity after 30 minutes. A farnesyl-derived isoprenoid diketone, 5,9,13-trimethyl-8,12-tetradecadiene-2,3-dione, at 93 microM caused a 94% reduction after 30 minutes. Other dicarbonyl compounds found to be effective against FTase in vitro were (+/-)-6-(camphorquinone-10-sulfonamido)-hexanoic acid, 4,4'-biphenyldiglyoxaldehyde, dehydroascorbic acid 6-palmitate, 2 oxododecanal, and phenylglyoxal. Higher concentrations of the alpha-dicarbonyl compound resulted in more rapid and more extensive inactivation. These findings demonstrate that alpha-dicarbonyl compounds targeted to FTase interfere with protein farnesylation in vitro and may lead to derivatives that have utility as chemotherapeutic agents. PMID- 11475910 TI - Synthesis and antitumour activity of 1H,3H-thiazolo[3,4-a]benzimidazole derivatives. AB - A series of 1H,3H-thiazolo[3,4-a]benzimidazoles were synthesized and tested for their in vitro antitumour activity against 60 human tumour cell lines. Some derivatives exhibited both tumour growth inhibition activity and cellular selectivity. In particular, compound 8c, the most active of the series, was very active towards all cell lines at concentrations ranging from 10(-7)-10(-5) M. Compound 4a, on the other hand, was highly selective against the CNS cancer cell line. PMID- 11475911 TI - Coumarin derivatives as protease-sensitive prodrugs. AB - To overcome the lack of selectivity of anticancer drugs toward malignant cells, the development of prodrugs, which could be activated selectively by tumour specific proteases is the goal of these studies. In this work tripartate prodrugs have been evaluated consisting of a carrier unit and a spacer group, which allows for intramolecular cyclisation while releasing the third component, the compound attached to the carboxylic acid moiety of the spacer group. As carrier units amino acids or peptides have been used, which are required for recognition by the protease. As the spacer unit the "trimethyl-lock"-spacer has been applied; as a model leaving group p-anisidine was attached to the carboxylic acid moiety. It was intended to test the compounds for their releasing rate of p-anisidine. Two of the evaluated compounds, 9b and 9h, were degraded with half-lives of 46 min at room temperature. However, the poor solubility in aqueous solutions proved the major disadvantage of the TML-based prodrugs. PMID- 11475912 TI - Participant supervision in co-therapy. AB - The present paper describes a model for supervision through co-therapy, developed in a training framework for interns of clinical psychology. The format presented includes a senior therapist and an intern as co-therapists. The model is conceptualized as one of participant supervision in psychodynamic psychotherapy. As such, the model is comprised of two elements. 1. Two therapists work together with one or more patient during a session. 2. The definition of the process as a supervisory situation, beyond the co-work in therapy of the two therapists. We offer the model as a significant and unique supplementary experience for both supervisor and supervise. The supervisory process is based on the principle of "reflection in action," and the actual participation of both partners in the clinical work, facilitating their mutual growth. PMID- 11475913 TI - Reliability and validity of the translated Psychotherapy Expectancy Inventory. AB - Matching treatment to therapy expectations is important. It increases treatment benefit and prevents dropout. No treatment expectation inventory existed in Hebrew, therefore a translation from English, of the Psychotherapy Expectancy Inventory-Revised, developed by Berzins, was undertaken. The questionnaire was given to 268 students twice, to 21 therapists and 107 parents of children referred to a child and adolescent clinic once. The translated questionnaire has high test-retest reliability, high internal consistency and significant criterion related validity-contrasted groups on advice, audience and relationship. Factor analysis yielded the four original scales, with the exception of two questions, which were moved to different scales. PMID- 11475914 TI - Psychotherapy and honoring parents. AB - This paper raises halachic issues regarding psychotherapy and honoring parents. A recognized halachic authority was asked to respond to a clinical case that involved the psychological treatment of a college student sexually abused by her father, where an important aspect of treatment focused on helping her discover and express her anger towards the offending parent instead of suppressing and internalizing it. Ramifications of the rabbi's ruling are summarized by an halachic scholar. PMID- 11475915 TI - Suicide during clozapine treatment: a case report. AB - The risk of suicide among patients with schizophrenia is greater than that in the general population. Although clozapine, along with its other significant features, has been reported to decrease suicidality and suicide rate among schizophrenia patients, physicians should remain vigilant for general risk factors of suicide in patients with schizophrenia during a treatment course and assess them carefully. Awareness of the effect of the illness, a sense of inadequacy to achieve goals, a change in the course of the illness, an improvement after relapse and a proposed discharge from the hospital in the near future are factors that should be noted especially within these general risk factors for suicide. In this case report, we presented a schizophrenia patient with several risk factors who committed suicide during clozapine treatment. Risk factors prominent in this case including increased awareness after improvement in chronic state, planned discharge in the near future, and the notion of reduced suicidality in schizophrenia patients during clozapine treatment are discussed in this article based on the literature and the case. PMID- 11475916 TI - LSD-induced Hallucinogen Persisting Perception Disorder treated with clonazepam: two case reports. AB - Benzodiazepines are recommended for the treatment of Hallucinogen Persisting Perception Disorder (HPPD), although it is unclear which may be more helpful. Two out-patients with LSD-induced HPPD were successfully treated with clonazepam. They had not responded to low potency benzodiazepines or low doses of classic antipsychotics. After clonazepam discontinuation they reported a marked improvement and only mild symptomatology which persisted during a six month follow-up period. High potency benzodiazepines like clonazepam, which has serotonergic properties, may be superior to low-potency benzodiazepines in the treatment of some patients with LSD-induced HPPD. PMID- 11475917 TI - The ethics of involuntary civil commitment. PMID- 11475918 TI - World Health Day on April 7, 2001 the theme is mental health. PMID- 11475919 TI - Alexithymia in patients with antisocial personality disorder in a military hospital setting. AB - We investigated the prevalence of alexithymic features and other psychometric correlates in patients diagnosed with antisocial personality disorder in a military hospital setting. Forty soldiers diagnosed with antisocial personality disorder in a general military hospital and 50 normal soldiers with no known medical or psychiatric disorder were assessed by sociodemographic data form, the Toronto Alexithymia Scale (TAS)-26 items, the Beck Depression Scale, the Beck Hopelessness Scale, the Brief Symptom Inventory and the State-Trait Anxiety Inventory. Antisocial patients showed significantly higher rates of unemployment, lower educational and socioeconomic status, higher rates of self mutilation, previous suicide attempts, substance abuse, history of incarceration and broken family bonds. The patient group also displayed significantly higher scores on alexithymia, depression, hopelessness and general psychological distress measures. Alexithymia was not associated with other psychological measures but was associated with socioeconomic and educational status. The failure in the socialization process of these patients may pave the way for an inability to identify and communicate their feelings. To draw a more definitive conclusion on this issue, a study which recruits ASPD patients from the community and compares them with a sociodemographically matched patient control group is necessary. PMID- 11475920 TI - A randomized trial of fluoxetine versus amitriptyline in musculo-skeletal pain. AB - In a preclinical study we found fluoxetine alone to induce a serotonin-mediated, dose-dependent antinociceptive effect in the mouse hot plate assay. In the present study we evaluated the clinical implication of these findings, comparing the efficacy of fluoxetine with that of amitriptyline for musculo-skeletal pain. Forty non-depressed patients, suffering from low back pain and whiplash associated cervical pain were enrolled in a randomized, six-week, "blind-rater" study, comparing the analgesic effect of amitriptyline (50-75 mgs/day) with that of fluoxetine (20 mgs/day). Twenty patients were randomly assigned to the amitriptyline group, and twenty to the fluoxetine group. Visual analogue and verbal rating scales were used for the assessment of pain intensity and pain relief. Thirty-five patients concluded the study. Moderate or good relief of pain was reported by 14 of the 17 patients (82%) in the amitriptyline group, and by 14 of the 18 patients (77%) in the fluoxetine group. The difference in responses between amitriptyline and fluoxetine was not statistically significant. In our study, fluoxetine relieved low back pain and whiplash associated cervical pain with efficacy similar to that of amitriptyline, offering an alternative for patients unable to tolerate the tricyclic antidepressants' side effects. PMID- 11475921 TI - Cooperation between mental health professionals and doctors in a Balint-oriented supervision group. AB - A Balint-oriented supervision group for physicians is described concentrating on the study of the patient-doctor relationship, the recognition and diagnosis of psychiatric problems, and the planning of psychiatric treatment. The group includes five general practitioners, a gynecologist, a dermatologist, a psychiatrist and a psychologist, who have met once a month for an hour over a period of 12 years. Interaction between the physicians and the mental health professionals is illustrated by two clinical examples. The group helps the physician recognize, tolerate and use his countertransference feelings, and facilitates the examination and treatment of patients suffering from psychiatric problems. In Balint-oriented group work, the focus can be moved from physical symptoms to include observation of the patient's emotional life and significant object relations, to the factors that are crucial for his psychological balance. This kind of holistic observation in the examination and treatment of psychiatric problems is as important as appropriate laboratory investigations in the diagnosis and care of physical diseases. PMID- 11475922 TI - [Exercise training in COPD]. AB - Rehabilitation for patients with chronic obstructive pulmonary disease (COPD) is well established and widely accepted as a means of enhancing standard therapy in order to alleviate symptoms and optimize function. The primary goal of rehabilitation is to restore the patient to the highest possible level of independent function. There is, however, no consensus on the optimal training modalities. Both walking and cycling improve exercise performance; there is substantial evidence that lower extremity exercise training should be included in rehabilitation programs for patients with COPD. PMID- 11475923 TI - [Cardiovascular rehabilitation]. AB - In industrialized countries, coronary artery disease is the leading cause of mortality. Cardiovascular rehabilitation is one of the most important components of secondary prevention and is mainly prescribed after acute myocardial infarction or coronary artery revascularization. The aims of rehabilitation are not only the improvement of functional capacity but also the correction of risk factors and assistance to social and professional reintegration. There are numerous indications, including not only patients with coronary artery disease but also patients with heart failure and those who underwent cardiac surgery or heart transplantation. Cardiac rehabilitation consists in 3 successive stages. The first is performed during hospital stay. The two others take place in an approved center of multidisciplinary cardiac rehabilitation. The intensity of exercise is determined for each individual patient from the results of stress testing. The beneficial effects are well established: reduction in morbidity and mortality, improvement of quality of life and of pulmonary, cardiac, muscular and metabolic parameters. Its efficacy is only maximal when patients continue performing exercise regularly. PMID- 11475924 TI - [Muscular exercise in the diabetic patient]. AB - Muscular exercise is recommended in patients with type 1 or type 2 diabetes mellitus and is part of the overall management, in combination with diet, oral antidiabetic agents and/or insulin. From a pathophysiological point of view, regular physical activity enhances insulin sensitivity, thus contributing to improve blood glucose control and to reduce cardiovascular risk factors associated to diabetes mellitus. From a practical point of view, however, muscular exercise in a diabetic patient requires special caution, in particular careful selection of the type of exercise, respect of metabolic and/or cardiovascular contraindications, and appropriate adjustment of current pharmacological/dietary treatment. PMID- 11475925 TI - [Sports and hypertension]. AB - Regular physical exercise belongs to the non pharmacological tools for the control of high blood pressure. When practising it almost daily at low intensity during 30 minutes, and mainly on a dynamic mode, blood pressure can decrease almost of the same order of magnitude as with an antihypertensive drug. In severe hypertension, blood pressure must be first controlled by drugs before starting the physical exercise training. An exercise test is preferable before exercise suggestion in sedentary people older than 40 years. In hypertensive people who enter sportive competition, diuretics and betablockers are forbidden. These agents can also reduce performance. PMID- 11475926 TI - [Echocardiography in athletes]. AB - Sporting activities and athletic training induce not only physiologic, but also morphologic changes. Dynamic training is associated with an enlarged left ventricle and a small increase in wall thickness. Athletes who perform predominantly static or isometric exercise have increased wall thickness with normal left ventricular internal diameter. Systolic and diastolic function are normal. Echocardiography is the method of choice to identify and follow the athlete's heart and to distinguish between physiologic and pathologic hypertrophy. PMID- 11475927 TI - [Sudden death in athletes]. AB - Sudden death is rare in the young athlete. The causes may vary. In the US, hypertrophic cardiomyopathy plays the predominant role whereas in Europe right ventricular arrhythmogenic dysplasia and atherosclerosis of the coronary arteries are more frequent. Other causes such as congenital anomalies of the coronary vessels, myocarditis, Marfan's disease, the long QT, the Brugada and the Wollf Parkinson-White syndromes exist, but are rare. Attentive preparticipation screening (clinical history and medical examination) is mandatory in all future young athletes. PMID- 11475928 TI - [Arrhythmias and sports activities]. AB - Arrhythmias do not systematically prevent athletic activities. Symptomatic bradyarrhythmias may first require pacemaker implantation. Usually occurring on normal heart, supraventricular tachycardias are benign. When responsible for palpitations, exercise intolerance or syncope, they may benefit from the use of beta-blockers, calcium channel blockers or most frequently radiofrequency catheter ablation. Ventricular arrhythmias with underlying cardiac disease do not allow intense and sustained physical activities. In case of symptomatic ventricular arrhythmias without cardiac disease, the use of beta-blockers, calcium channel blockers or radiofrequency catheter ablation usually permits sport practice. PMID- 11475929 TI - [Exercise in children with chronic disease]. AB - Regular physical exercise is gaining acceptance as a prescription for the prevention, management and rehabilitation of diseases occurring in the pediatric and adolescent age group. Exercise is generally used to increase the physical capabilities of these young patients, but, at times, it can directly influence the evolution of the condition. However, in children with chronic diseases, enhanced physical activity can also be deleterious. A well structured exercise program should be implemented. This paper will focus on the role of exercise in different diseases including cardiovascular, pulmonary, neuromuscular disease, diabetes and obesity. PMID- 11475930 TI - [Exercise-induced asthma, a complex pathology. New answers, new questions]. AB - This is a short review for general practitioners about the clinical features of exercise induced asthma (EIA), sometimes unusual in presentation, chiefly among children. Some most currently accepted theories explaining the disease are recalled. The ambiguous relationship between asthma and exercise is pointed out: Exercise is a major comportent of asthma treatment but asthma seems more and more a prominent complication of high level sport practice in some disciplines. The need for a clear international definition of EIA is outlined as bronchial hyperactivity alone does not seem a clear-cut criterion for asthma in the general population as well as in athletes. Some useful drugs for the control of EIA are mentioned. A final question: EIA an under- or overdiagnosed disease? PMID- 11475931 TI - [Cutaneous infections contracted during sports and recreational activities]. AB - Going in for sports is at increased risk for cutaneous infections that can be viral, bacterial, mycotic and parasitic in nature. The specific conditions of sports practice have a major influence on their occurrence. Other factors such as age, gender and genetic predisposition can also play a role. The cutaneous infections should be the target of preventive measures. To assume the worse, they should be recognized and treated without delay in order to avoid the more or less prolonged arrest of the sports activity and the disease transmission to partners. PMID- 11475932 TI - [Overtraining in endurance athletes]. AB - Optimal training results in exercise-recovery harmonious balance. High frequency and volumic training, associated with insufficient recovery may induce overtraining. It depends on the kind of exercise but occurs mainly in endurance sports. The high frequency of upper respiratory track infection in endurance sportsmen results from a temporary immunosuppression state, possibly associated with overtraining. Many immune and haematological parameters have been associated with overtraining, but none seemed really efficient for diagnosis. Hormonal parameters, despite contradictions in studies, appear to give a good insight of athlete's staleness and are more predictive of overtraining. Actually, overtraining detection needs a combination of different parameters, and cannot sustain on isolated factors. PMID- 11475933 TI - [Muscle injuries in athletes: pathogenesis, diagnosis and treatment]. AB - All muscles can be injured during sports activities, certain specific muscles in the lower extremities are particularly vulnerable. The spectrum of these injuries is wide. Muscle strain injuries frequently involve polyarticular muscles which contain a high percentage of fast-twitch fibres (Type II). Disruption of muscle fibres occurs near the myotendinous junction. In most cases, the diagnosis can be based upon history and physical examination. Ultrasound and Magnetic Resonance Imaging can be useful to determine the severity of a specific injury. The treatment of these injuries is basically conservative; surgery is rarely indicated. Prevention of these injuries is currently still an important issue. PMID- 11475934 TI - [Isokinetic exercise and sports injuries]. AB - The assessment of human muscle strength is of considerable value in various pathological states with modifications of the performance level. The original concept of isokinetic dynamometry provides a precise and reliable quantification of strength. Identification of underlying deficits in bilateral and reciprocal muscle group relationships plays a great role in the treatment of anterior cruciate ligament (ACL) reconstruction and muscle injuries. The value of this type of assessment for prevention in uninjured subjects practicing sports at risk is suggested. Rehabilitative use of isokinetic includes muscle strengthening and tendinitis treatment by means of eccentric exercises. PMID- 11475935 TI - [Stress fractures and sports]. AB - Stress fractures are very frequent injuries which account for 10% of all consultations in sports medicine, in a majority of cases affecting the lower limbs in young adults. They can be defined as a modification of the bone structure not necessarily associated with cortical infraction which occurs in the absence of injury. They happen as a result of an imbalance between the bone's capacity for adaptation and the overloading of mechanical constraints applied. This imbalance is adjusted by different favourable factors thus explaining the individual threshold to external demands. In theory, any bone might be damaged although more frequently they are to be found at the level of the tibia or the second and third metatarsals. The clinical picture is dominated by a symptomatology of pain which appears progressively strictly mechanical at the development stage. Further medical examination will include an x-ray (late diagnosis which often gives false-negative results) but, more importantly, a scintigraphy (which is very sensitive at an early stage but which is not specific enough). More recently, MRI has been a preferred means of detection. The treatment consists of obligatory segmented rest for a period of between two and six weeks. The intensity and length of treatment vary in function of the location of the injury and the reoccurrence of pain when the offending activity is restarted again. Rest is of utmost importance in order to avoid an evolution towards a true fracture which would result in long-term isolation from sport and social activities. PMID- 11475936 TI - [Tendon pathologies in veteran athletes]. AB - Tendonitis and tendons ruptures are mainly seen above 40 years of age, especially in sport practice. We discuss the physiopathology, diagnosis and treatment. PMID- 11475937 TI - [Entrapment neuropathies in sports medicine]. AB - The more frequent entrapment neuropathies related to sport are described: thoracic outlet syndrome in aquatic athletes and pitchers, long thoracic neuropathy in tennis players, suprascapular neuropathy in volleyball and tennis players, ulnar nerve entrapment at the elbow in pitchers and at the wrist in cyclists, Morton's syndrome in runners and dancers. PMID- 11475938 TI - [Muscle electric stimulation in sports medicine]. AB - Neuromuscular electrical stimulation (NMES) is frequently used to strengthen the normal muscle. In medicine, several investigators have highlighted the value of NMES when treating amyotrophy related to immobilization. The efficacy of NMES of the quadriceps during rehabilitation following anterior cruciate ligament surgery has been well documented. These electrical stimulation programs are composed of stimulation--rest cycles and are performed with the aid of percutaneous electrodes situated in regard to muscle motor points. The stimulation frequency is chosen to induce tetanic contractions and is generally situated between 50 and 100 Hz. During NMES, the trainer or the physiotherapist can accurately modulate a large number of parameters but the sensory discomfort linked to the current intensity remains the major limitation for strength improvement. In the context of rehabilitation, the association of NMES with voluntary exercises is superior to the isolated utilization of the classic program composed of isometric voluntary contractions. To explain this advantage of associating NMES with voluntary contractions, it has been hypothesized that the electrically induced contractions impose specific patterns of muscle recruitment and a particular metabolic solicitation. PMID- 11475939 TI - [Guidelines for good practice of sports activities]. AB - The present closing article summarizes some guidelines for the good practice of physical activities in order to develop and maintain cardiorespiratory and muscular fitness, and flexibility. Advice is given regarding the recommended quantity and quality of exercise in term of intensity, duration and frequency of training with the aim to optimize the risk/benefit ratio for health, in both aerobic endurance and resistance exercises. The crucial role of an appropriate warm-up and cool-down period, which would include flexibility exercises, is also emphasized. Finally, some practical examples illustrate this vademecum of physical activities. PMID- 11475940 TI - The new generation of GABA enhancers. Potential in the treatment of epilepsy. AB - gamma-Aminobutyric acid (GABA) is considered to be the major inhibitory neurotransmitter in the brain and loss of GABA inhibition has been clearly implicated in epileptogenesis. GABA interacts with 3 types of receptor: GABAA, GABAB and GABAC. The GABAA receptor has provided an excellent target for the development of drugs with an anticonvulsant action. Some clinically useful anticonvulsants, such as the benzodiazepines and barbiturates and possibly valproic acid (sodium valproate), act at this receptor. In recent years 4 new anticonvulsants, namely vigabatrin, tiagabine, gabapentin and topiramate, with a mechanism of action considered to be primarily via an effect on GABA, have been licensed. Vigabatrin elevates brain GABA levels by inhibiting the enzyme GABA transaminase which is responsible for intracellular GABA catabolism. In contrast, tiagabine elevates synaptic GABA levels by inhibiting the GABA uptake transporter, GAT1, and preventing the uptake of GABA into neurons and glia. Gabapentin, a cyclic analogue of GABA, acts by enhancing GABA synthesis and also by decreasing neuronal calcium influx via a specific subunit of voltage-dependent calcium channels. Topiramate acts, in part, via an action on a novel site of the GABAA receptor. Although these drugs are useful in some patients, overall, they have proven to be disappointing as they have had little impact on the prognosis of patients with intractable epilepsy. Despite this, additional GABA enhancing anticonvulsants are presently under development. Ganaxolone, retigabine and pregabalin may prove to have a more advantageous therapeutic profile than the presently licensed GABA enhancing drugs. This anticipation is based on 2 characteristics. First, they act by hitherto unique mechanisms of action in enhancing GABA-induced neuronal inhibition. Secondly, they act on additional antiepileptogenic mechanisms. Finally, CGP 36742, a GABAB receptor antagonist, may prove to be particularly useful in the management of primary generalised absence seizures. The exact impact of these new GABA-enhancing drugs in the treatment of epilepsy will have to await their licensing and a period of postmarketing surveillance. As to clarification of their role in the management of epilepsy, this will have to await further clinical trials, particularly direct comparative trials with other anticonvulsants. PMID- 11475941 TI - Psychogenic excoriation. Clinical features, proposed diagnostic criteria, epidemiology and approaches to treatment. AB - Psychogenic excoriation (also called neurotic excoriation, acne excoriee, pathological or compulsive skin picking, and dermatotillomania) is characterised by excessive scratching or picking of normal skin or skin with minor surface irregularities. It is estimated to occur in 2% of dermatology clinic patients and is associated with functional impairment, medical complications (e.g. infection) or substantial distress. Psychogenic excoriation is not yet recognised in the DSM. We propose preliminary operational criteria for its diagnosis that take into account the heterogeneity of behaviour associated with psychogenic excoriation and allow for subtyping along a compulsivity-impulsivity spectrum. Psychiatric comorbidity in patients with psychogenic excoriation, particularly mood and anxiety disorders, is common. Patients with psychogenic excoriation frequently have comorbid disorders in the compulsivity-impulsivity spectrum, including obsessive-compulsive disorder, body dysmorphic disorder, substance use disorders, eating disorders, trichotillomania, kleptomania, compulsive buying, obsessive compulsive personality disorder, and borderline personality disorder. There are few studies of the pharmacological treatment of patients with psychogenic excoriation. Case studies, open trials and small double-blind studies have demonstrated the efficacy of selective serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitors in psychogenic excoriation. Other pharmacological treatments that have been successful in case reports include doxepin, clomipramine, naltrexone, pimozide and olanzapine. There are no controlled trials of behavioural or psychotherapeutic treatment for psychogenic excoriation. Treatments found to be effective in case reports include a behavioural technique called 'habit reversal'; a multicomponent programme consisting of self monitoring, recording of episodes of scratching, and procedures that produce alternative responses to scratching; and an 'eclectic' psychotherapy programme with insight-oriented and behavioural components. PMID- 11475942 TI - Psychiatric disorders associated with Cushing's syndrome. Epidemiology, pathophysiology and treatment. AB - Cushing's syndrome is caused by a chronic excess of glucocorticoids. A number of psychiatric and psychological disturbances may be associated with the condition, regardless of its aetiology. Major depression is the most common comorbid disorder. Other psychopathological aspects of Cushing's syndrome in adults include mania, anxiety disorders and cognitive dysfunction. The presence of depression connotes a severe clinical presentation and, in patients with hypothalamic-pituitary forms of Cushing's syndrome, is prognostically useful. Inhibitors of corticosteroid production (e.g. ketoconazole, metyrapone, aminoglutethimide), rather than antidepressant drugs, are generally successful in relieving depressive symptoms, as well as other disabling symptoms. These drugs can be used to control symptoms prior to surgical treatment of Cushing's syndrome. Long-standing hypercortisolism may cause some degree of irreversible pathological damage and induce highly individualised affective responses based on each patient's psychological assets and liabilities. As a result, upon normalisation of cortisol levels, treatment may still be required, and should encompass both psychotherapeutic strategies (particularly cognitive-behavioural therapies that have been found to be effective in affective disorders) and psychotropic drug treatment [antidepressants such as tricyclic agents and selective serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitors]. In patients with severe anxiety, benzodiazepines (e.g. clonazepam in small doses) may also be helpful. PMID- 11475943 TI - Pharmacodynamic-tolerability relationships of cholinesterase inhibitors for Alzheimer's disease. AB - According to the cholinergic hypothesis, the impairment of cognitive function and the behavioural disturbances that affect patients with Alzheimer's disease are mainly due to cortical deficiencies in cholinergic transmission. Numerous cholinesterase inhibitors have been investigated for treatment of this disease, the rationale being to support the cholinergic system by blocking the degradation of acetylcholine released from presynaptic neurons. These drugs can be classified as reversible (tacrine, donepezil and galantamine), pseudo-reversible (physostigmine, eptastigmine and rivastigmine) or irreversible (metrifonate) enzyme inhibitors. This article reviews efficacy and tolerability results from 6 month placebo-controlled studies of 7 cholinesterase inhibitors: tacrine (80 to 160 mg/day), donepezil (5 to 10 mg/day), rivastigmine (1 to 12 mg/day), metrifonate (30 to 80 mg/day), eptastigmine (30 to 60 mg/day), physostigmine (30 to 36 mg/day) and galantamine (8 to 32 mg/day). All these agents have demonstrated a statistically significant, although modest, effect versus placebo on the cognitive and global performance of patients with Alzheimer's disease. Dramatic clinical response has been seen in only 3 to 5% of patients. There are no major differences in terms of efficacy between the different drugs. The mean difference between drug and placebo effects on standardised psychometric scales is about 2 to 4 points on the cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-Cog; a 70-point cognitive scale) and 0.2 to 0.5 points on the Clinician's Interview-Based Impression of Change with Caregiver Input (CIBIC Plus; a 7-point global scale), or 5 to 14% of the average value of the scales. The most common adverse effects observed after administration of cholinesterase inhibitors are nausea, vomiting, diarrhoea, dizziness, asthenia and anorexia, all symptoms linked to cholinergic overstimulation. These effects are dose related and largely depend on the degree of cholinesterase inhibition. Also important is the rate of onset of cholinesterase inhibition, which depends on the kinetics of enzyme inhibition, the presence and rate of titration, and the pharmacodynamic peak-to-trough fluctuations. A model predicting the incidence of nausea based on acetylcholinesterase inhibition and the half-life of acetylcholinesterase recovery is proposed. In conclusion, cholinesterase inhibitors are the only pharmacological agents proved to be effective for the treatment of Alzheimer's disease in large, long term, double-blind, placebo-controlled trials. While the efficacy of different cholinesterase inhibitors is similar, their tolerability profiles differ. For example, the incidence of nausea (in excess of that seen with placebo) at cognitively effective dosages ranges from 1% with eptastigmine 60 mg/day to 53% with physostigmine 30 mg/day. Differences in tolerability profile may be due to the extent of peripheral acetylcholinesterase inhibition needed to reach clinical efficacy. Other contributing pharmacodynamic factors are the rate of onset of and fluctuations in acetylcholinesterase inhibition at steady state. PMID- 11475944 TI - Smoking cessation in women. Special considerations. AB - Women may be at relatively greater risk of smoking-related diseases than men but tend to have less success than men in quitting smoking. The failure of most outcome studies to report results by gender and the lack of statistical power for detecting significant gender differences currently do not allow for many firm conclusions to be drawn about smoking cessation rates in women, but several trends warrant attention and further study. First, the difference in cessation rates for women versus men may be even greater in trials of nicotine replacement therapies (NRT). This suggests that women benefit less from NRT relative to men, although this difference may depend on the particular form of NRT (e.g. inhaler versus gum). On the other hand, some non-NRT medications may reverse the poorer outcome of women, producing quit rates in women comparable with those in men. Gender differences in outcome, as well as overall success rates, with NRT and some of the non-NRT medications appear to be enhanced when treatment includes substantial behavioural counselling. However, while several of the non-NRT medications may be particularly appropriate to consider for treating women trying to quit smoking, adverse effects may limit widespread use of some of these drugs, such as clonidine and naltrexone. Thus, even if the gender differences in outcome with NRT versus non-NRT drugs are confirmed in further research, such findings do not necessarily justify limiting NRT use in women, because such treatment is clearly effective and is likely to be safer and more readily available than non NRT medications. Nevertheless, study of the mechanisms by which some non-NRT drugs are effective in women may aid our understanding of factors that are more influential in smoking behaviour in women than in men. Secondly, smoking cessation treatment for women must address several other issues that often emerge, and these are most likely to require behavioural counselling that is tailored to these problems. These issues include concern about bodyweight gain, restrictions on medication use in pregnant smokers, variability in mood and withdrawal as a function of menstrual cycle phase, harnessing social support to foster abstinence, and the possibility that smoking-associated environmental cues may be more influential in smoking behaviour in women than men. Greater attention to gender differences in clinical trial outcomes and to addressing concerns of women smokers may aid in the development of substantially improved smoking cessation interventions for women. PMID- 11475946 TI - Guidelines for the evaluation and management of traumatic dental injuries. PMID- 11475945 TI - Sleep abnormalities during abstinence in alcohol-dependent patients. Aetiology and management. AB - Virtually every type of sleep problem occurs in alcohol-dependent patients. Typically, these individuals take a longer time to fall asleep and show decreased sleep efficiency, shorter sleep duration and reduced amounts of slow wave sleep when compared with healthy controls. Their sleep patterns are fragmented, and the typical time course of electroencephalogram (EEG) delta wave activity is severely disrupted. The amount of rapid eye movement (REM) sleep may be reduced or increased. Sleep changes can persist during months or years of abstinence, and recent studies indicate that certain alterations in sleep architecture, as well as subjective sleep complaints, predict relapse to alcoholism. The mechanisms of action of short and long term alcohol administration on sleep are incompletely understood. They may arise from an interaction with gamma-aminobutyric acid (GABA), serotonin (5-hydroxytryptamine; 5-HT), adenosine or other neurotransmitter systems. While only a few pharmacological and nonpharmacological strategies to improve or normalise disturbed sleep in individuals who have recovered from alcoholism have been studied, the use of benzodiazepines, other hypnosedatives or selective serotonin reuptake inhibitors is not recommended. Therapies include sleep hygiene, bright light therapy, meditation, relaxation methods, and other nonpharmacological approaches. Further studies are needed to clarify the relationship between sleep, sleep abnormalities and alcoholism, and to establish new approaches to improve sleep in alcohol-dependent patients and to prevent withdrawal reactions that affect sleep during abstinence. PMID- 11475947 TI - Healing of 208 intra-alveolar root fractures in patients aged 7-17 years. AB - This retrospective study consisted of 208 root-fractured, 168 splinted and 40 not splinted incisors in young individuals (aged 7-17 years) treated in the period 1959-1973 at the Pedodontic Department, Eastman Institute, Stockholm. Clinical and radiographic analyses showed that 69 teeth (33%) had developed hard tissue (fusion) healing of fragments. Interposition of periodontal ligament (PDL) and bone between the fragments was found in 17 teeth (8%). Interposition of PDL alone was found in 74 teeth (36%). Finally, non-healing with pulp necrosis and inflammatory changes between fragments was seen in 48 teeth (23%). Various clinical factors were analyzed for their relationship to the healing outcome with respect to healing/no healing and type of healing (hard tissue versus interposition of bone and/or PDL). Immature root and positive pulp sensitivity at time of injury was found to be significantly related to both pulp healing and hard tissue repair of the fracture. The same applied to concussion or subluxation of the coronal fragment compared to luxation with displacement (extrusive or lateral luxation). This relation was also represented by the variable millimeter diastasis between fragments before and after repositioning. Repositioning appeared to enhance the likelihood of both pulp healing and hard tissue repair. A positive effect of splinting, splinting methods (cap splints or orthodontic bands with an arch wire) or splinting periods could not be demonstrated on either pulp healing or type of healing (hard tissue versus interposition of bone and/or PDL). In conclusion, the findings from this retrospective study have cast doubts on the efficacy of long-term splinting and the types of splint used for root fracture healing. It is suggested that the role of splinting and splinting methods be examined in further studies. PMID- 11475948 TI - Efficacy of laser Doppler flowmetry for the diagnosis of revascularization of reimplanted immature dog teeth. AB - This study was performed to assess if laser Doppler flowmetry (LDF) is an improved method for the detection of revascularization of replanted teeth. Teeth were extracted and reimplanted under different experimental conditions. LDF readings were taken before extraction and weekly for 3 months. In control teeth, LDF baseline readings were taken and then repeated after the apical blood vessels were cut surgically. At the end of 3 months it was determined radiographically and histologically whether revascularization had occurred, i.e. vitality had returned. RESULTS: LDF readings correctly predicted the pulp status (vital vs. non-vital) in 83.7% of the readings. 73.9% (17 of 23) were correct for the vital teeth and 95% (19 of 20) were correct for the non-vital teeth. Fisher's exact test (2-tail) indicated that there was no significant association between the efficacy of LDF and tooth type (P = 0.166), although P2 was the least accurate tooth tested. Wilcoxon's matched-pair signed rank test demonstrated that in the revascularized (vital) teeth, the flux value between the baseline and week 2 dropped significantly (P = 0.0001), increased significantly from week 2 to week 4 (P = 0.0001) and then decreased steadily until week 12. However, at week 12 the flux was still significantly higher than at week 2 (P = 0.010). In the teeth that failed to revascularize, the flux value dropped significantly by weeks 1 and 2 (P = 0.004 and P = 0.0001, respectively). Flux values did not increase from week 2. A Fast Fourier Transform (FFT) analysis confirmed a pulse of dominant frequency of 2 Hz in the teeth that returned to vitality and the lack thereof in those that stayed non-vital. One tooth in which the flux value evaluation indicated a non vital tooth but the radiographic/histologic findings showed vital (false negative) possessed a pulse of dominant frequency and proved by this method to have successfully revascularized. PMID- 11475949 TI - Dental trauma management awareness of Singapore pre-school teachers. AB - The purpose of this study was to assess the awareness of pre-school teachers concerning the management of traumatized teeth. A questionnaire survey of teachers' backgrounds, attitudes/practices and knowledge was distributed to all teachers who attended a dental education program organized by the Singapore Dental Health Foundation. Of a total of 291 teachers surveyed, 29% had received tertiary education, while the rest (71%) had received secondary education. The mean teaching experience was 6.8 +/- 6.9 years. About a quarter had previous experience in handling dental trauma. Sixty-three percent admitted having no knowledge of dental trauma; 79% were unsatisfied with their level of knowledge in this area; 95% were keen to have further education in dental trauma; 65% thought dental trauma emergency should be dealt with as soon as possible. Concerning knowledge, during-office hour emergency services were more familiar (84%) than after-office hour emergency services (15%), as was the concept of management of avulsed teeth (71%) compared to that of fractured teeth (51%). Knowledge about optimal storage media for avulsed permanent teeth was especially poor--being as low as 15%. Using multiple logistic regression analysis, it was found that teaching experience significantly influenced the respondents' self-assessed knowledge and their level of satisfaction with their knowledge (P = 0.012). Teachers with more teaching experience had better knowledge about the replantation of permanent teeth (P = 0.003). It is recommended that public education targeted at teachers should be carried out to increase dental trauma management awareness. PMID- 11475950 TI - Lay knowledge of physical education teachers about the emergency management of dental trauma in Hong Kong. AB - The purpose of this study was to investigate the knowledge of a group of physical education (PE) teachers about the emergency management of dental trauma in Hong Kong. A postal questionnaire was sent to PE teachers in 100 randomly selected secondary schools in Hong Kong. A total of 166 teachers from 65 schools responded. Results show that a majority of the respondents have received formal teaching training (98.8%). While all but one teacher had received formal first aid training, only eight of them recalled that they had received advice on the management of dental trauma. Concerning the management of tooth fracture, 118 respondents (71.1%) gave an appropriate answer. In contrast, only 29 teachers (17.5%) were able to indicate the appropriate management for an avulsed tooth. Over 60% (102) of the respondents indicated it was "very urgent" to seek professional assistance if a permanent tooth has been avulsed, but they had little knowledge of the correct procedures for replanting or transporting avulsed teeth. Only 15 respondents (9%) pointed out that milk was the medium of choice for transporting avulsed teeth. Finally, over 90% (157) of respondents indicated that they had never received advice on the emergency procedures for the management of dental avulsion. The present study showed that the level of lay knowledge of management of dental injuries in a selection of PE teachers in Hong Kong is inadequate and educational campaigns are necessary to improve their emergency management of dental injuries. PMID- 11475951 TI - Calcium hydroxide retention in wide root canals with flaring apices. AB - Calcium hydroxide (CH) is used to induce apexification in immature, pulpless teeth with wide root canals and flaring apices. Three placement methods of CH were compared in vitro, for their ability to enhance CH retention in the canal. Barium sulfate (BS), added as radiopacifier, was studied for its effects on the radiographic follow up of CH retention, as well as on the ability to detect voids in the CH. Calcium hydroxide filled teeth were kept in phosphate buffered saline containing tubes in a shaker water bath. Radiographs were taken weekly to follow the retention or loss of the material; they were scanned and stored as digitized images. Quantitative evaluations were done on these images, using a computerized gray scale. Condensed CH was better retained in root canals than either lentulo placed paste or commercial injected paste. Barium sulfate in the CH paste enabled better detection of voids in the paste filling the canal, however, it obscured the disappearance of CH from the canal, due to a residual radiopacity effect. The results supported (a) condensation of calcium hydroxide as a preferred method that improves its retention in wide root canals, and (b) discontinuing the use of barium sulfate as a radiopacifier in apexification procedures. PMID- 11475952 TI - Decoronation of an ankylosed tooth for preservation of alveolar bone prior to implant placement. AB - A 12-year-old patient sustained avulsions of both permanent maxillary central incisors. Subsequently, both teeth developed replacement resorption. The left incisor was extracted alio loco. The right incisor was treated by decoronation (removal of crown and pulp, but preservation of the root substance). Comparison of both sites demonstrated complete preservation of the height and width of the alveolar bone at the decoronation site, whereas the tooth extraction site showed considerable bone loss. In addition, some vertical bone apposition was found on top of the decoronated root. Decoronation is a simple and safe surgical procedure for preservation of alveolar bone prior to implant placement. It must be considered as a treatment option for teeth affected by replacement resorption if tooth transplantation is not feasible. PMID- 11475953 TI - The evolution of gait analysis: a treatment decision-making tool. AB - The development of analytical techniques to define the abnormalities contributing to pathological gait patterns in human locomotion is complex. From the earliest attempts to draw static figures of the human form, physicians and scientists have devised unique paradigms to analyze various body segments in motion. The earliest studies were founded upon the discoveries of anatomy and physiology in the Renaissance. All subsequent advances have developed on that foundation. The discoveries of photography, electromyography, and the computer have led to today's advanced technology. Abnormalities of human locomotion can now be defined with accuracy in three dimensions and sufficient speed to be viable in a clinical setting. The objective documentation of gait through clinical gait analysis has not only gained for the physician a better understanding of the pathomechanics of gait abnormalities; it has become a valuable tool in treatment decision-making and evaluation. In treatment decision-making, gait analysis is used routinely to determine specific surgical procedures in patients of all ages with neuromuscular and other disorders. In treatment evaluation, gait analysis is becoming more relevant with the recent emphasis on outcome measures as a justification for insurance coverage. PMID- 11475954 TI - Cutaneous leishmaniasis in an Italian woman: case report. AB - Leishmaniasis is a sandfly-borne disease caused by a protozoan. The typical lesion of cutaneous leishmaniasis first appears as an erythematous papule at the site of inoculation, increases slowly in size, develops raised borders, and eventually ulcerates. The pentavalent antimony compounds continue to be a mainstay of therapy. We describe an Italian patient with an enlarging facial plaque that was found to be caused by leishmania and discuss the toxicity associated with therapy. PMID- 11475955 TI - Confidentiality laws and the physician. PMID- 11475956 TI - On privacy. PMID- 11475957 TI - "Hypoplastic left heart syndrome: factors influencing therapeutic choice". PMID- 11475958 TI - "Images in clinical medicine". PMID- 11475959 TI - "Primary care: strategies for survival in the era of managed care". PMID- 11475960 TI - [Clinical signs, strategies of treatment and complications of postnecrotic pancreatic cyst]. AB - There were examined 130 patients with postnecrotic pancreatic cyst (PNPC). In 16.2% of them the cause of the cyst formation was not established. Operative intervention performance is indicated after establishment of the PNPC diagnosis before occurrence of complications. PMID- 11475961 TI - [Surgical treatment of chronic pancreatitis with ductal hypertension complicated by posterior pancreatic pseudocyst]. AB - The experience was summarized concerning the treatment of 5 patients with chronic pancreatitis and ductal hypertension, complicated by retropancreatic pseudocyst. One-stage internal draining of pseudocyst and the pancreatic duct, using the method elaborated in the clinic, was performed. There were no postoperative complications. Late follow-up result is good. PMID- 11475962 TI - [Posttraumatic retroperitoneal hematoma in injured persons with severe closed multiple and combined abdominal trauma]. AB - The experience of treatment of 69 injured persons with posttraumatic retroperitoneal hematoma suffering severe multiple combined abdominal trauma was analyzed. Application of the classification proposed permits to formulate diagnosis and to choose the tactic of treatment correctly. The intraoperative tactics algorithm was elaborated. It promotes the correct analysis of intraoperative findings and reduction of the diagnostic mistakes frequency as well. In the presence of vast defect, making impossible to suture over the parietal peritoneum, extraperitonization using cerebral dura mater was done. Operative intervention was concluded by drainage with subsequent laserotherapy. PMID- 11475963 TI - [Application of enteral detoxication and decontamination in acute ileus]. AB - According to the experimental investigation data, conducted in 16 mongrel dogs in an acute ileus (AI), there are observed the colonization of proximal parts of small intestine (SI) by aerobic and anaerobic microorganisms, the power dependent processes of inhibition and activation of peroxidal oxidation of lipids in her mucosa, causing the disorders of her morphology. Conduction of intraoperative flowing intestinal lavage and the abdominal cavity sanation using betadin, diluted in 1:100 ratio, have promoted the pathological microorganisms quantity reduction and stimulation of power dependent processes in the SI mucosa. Application of enteral detoxication and decontamination in 86 patients with AI during performance and after the operation have permitted to reduce the intoxication severity, to restore the motor, evacuational, absorbtive function of SI, the postoperative complications frequency. PMID- 11475964 TI - [The evaluation of the external respiratory function in patients in late follow up period after pneumonectomy]. AB - The external respiration function (ERF) indexes were studied in 215 patients in 25-40 years after performance of pneumonectomy. In 55.3% patients ventilation indexes have exceeded the due values by 50% for two lungs, in 8.4%--significant disorders of the ERF indexes were noted in connection with an early and late complications occurrence and with presence of specific and metaphthisic changes in a single lung as well. PMID- 11475965 TI - [Ocular hydrodynamics in patients with occluding and stenosing disease of main cerebral vessels after reconstructive surgery of internal carotid and vertebral arteries]. AB - Ocular hydrodynamics was investigated using the tonography method in 58 patients, to whom reconstructive operations on the affected vessels were performed. After reconstructive operation on vertebral arteries ocular hydrodynamics didn't change significantly, on a carotis interna (ACI)--the positive effect was revealed on the side of the operation performed, and if the cerebrum arterial circle function was preserved--on the contralateral side as well. If extra- and intracranial parts of ACI were affected after conduction of reconstructive operation and restoration of the artery's passability occurrence of "ocular reperfusional syndrome" is possible, manifested by intraocular pressure raising due to excessive raising of the aqueous humor production. PMID- 11475966 TI - [Surgical treatment of bronchogenic and enterogenic mediastinal cysts complicated by intrathoracic compression of respiratory tracts in children]. AB - In the clinic there were operated on 8 patients with bronchogenic and enterogenic mediastinal cyst, complicated by intrathoracic compression of respiratory ways. In 2 children postoperative complications occurred. In presence of secondary tracheomalacia, caused by tracheal compression by bronchogenic cyst, aortopexy was done in conjunction with the cystic formation excision. All the patients survived. In 6 children satisfactory result was noted, in 2--relatively satisfactory one. PMID- 11475967 TI - [Internal bronchial studies in children pulmonary diseases]. AB - Bronchologic investigation of the inborn failures of development and acquired pulmonary diseases were studied in 148 children. There were done 313 endobronchial investigations. Peculiarities of course of nonspecific pulmonary diseases were studied basing on retrospective analysis of the case histories and ambulatory cards. PMID- 11475968 TI - [Immediate results of treatment of patients with rectal cancer using neoadjuvant chemotherapy and radiotherapy]. AB - Immediate results of treatment of 103 patients with cancer of the proximal part of rectum were analyzed. In 54 patients there was applied in complex of treatment neoadjuvant chemotherapy: intraabdominal infusion of fluorouracil (0.6 g/m2, 4-6 g dose for the course) in combination with gamma-therapy (20 Gr) with subsequent infusion of cisplatinum (60 mg/m2), to 49 patients only surgical treatment was done. Complex treatment, conducted according to the proposed method, didn't worsen the conditions of the operation performance, didn't increase intraoperative blood loss and frequency of intra- and postoperative complications, didn't change their structure and didn't worsen the immediate result of operation. PMID- 11475969 TI - [Efficacy and perspective of application of preparation based on hydrogel and xerogel of methyl-silicic acid in patients with the intestinal malignancy]. AB - There was studied the influence of medicinal preparations, created on the base of methylsiliconorganic matrix, on immediate and late follow-up result of treatment of more than 2000 patients with the digestive channel malignancy. High efficacy of application in complex of preoperative preparation of immobilized antibiotics and cytostatics, enterosorpent enterosgel for the purulent-inflammatory complications prophylaxis, lowering of bilirubin level in tumoral obturative jaundice, and in intraoperative usage for the tumor and metastases recurrences prophylaxis. PMID- 11475970 TI - [Treatment of colonic melanoma]. AB - The results of treatment of 5 patients with colonic melanoma are presented. Fine needle biopsy was used for morphological verification diagnosis. It was established that the patients with colonic melanoma are hospitalized in specialized department, as a rule, with nonopertable tumor. In 1 patient explorative laparotomy was done. Conduction of radical operative intervention was impossible because of presence of multiple metastases in liver. In 4 patients conservative treatment was performed. It was not possible to perform radical operation because of the process spreading. The patients life span after conduction of conservative therapy have constituted 14.5 mo at average. PMID- 11475971 TI - [Results of operative treatment of the thyroid gland cancer in the Krivorog region]. AB - Results of treatment of the thyroid gland cancer in 251 patients were analyzed. PMID- 11475972 TI - [Application of carbon sorbents in the treatment of experimental peritonitis]. AB - Comparative estimation of efficacy of sorpents SKN-1K, KAU, AU-L in experimental peritonitis was performed. Most favourable effect was observed after performance of hemosorption using AU-L. Sorpent KAU caused very pronounced detoxicational effect, but while its application additional correction of the glucose level is necessary, and that's why it is contraindicated in hypoglycemia. SKN absorbs toxic products least actively, but corrects the carbohydrate metabolism indexes better than KAU. PMID- 11475973 TI - [Restoration of blood circulation in reperfusion of ischemic tissues]. AB - In experiment there were studied changes of the tissue blood circulation and microcirculation during postischemic reperfusion of skeletal muscles with various density of capillaries. It was established that reactive hyperemia, caused mainly by enhancement of extracapillary tissue blood flow. While persistence of durable arterial hypertension the capillaries quantity and potential volume of microcirculatory bed are reducing, negatively influencing the blood circulation restoration process during postischemic reperfusion. In chronic ischemia the potential volume of microcirculation is increasing, securing complete restoration of blood circulation in postischemic reperfusion. PMID- 11475974 TI - [Prognostic dynamics of the main urgent surgical diseases in the last 13 years and its social medical significance]. PMID- 11475975 TI - [Influence of vagotomy on pancreatic blood flow in surgical treatment of gastroduodenal ulcer complicated by hemorrhage]. AB - In 56 patients with bleeding duodenal ulcer the influence of truncal, selective and selective proximal vagotomy on pancreatic blood flow was studied, basing on the intraoperative rheopancreatography data. The advantages of selective proximal vagotomy in correction of the disorders revealed were proved. PMID- 11475976 TI - [Application of x-ray-assisted surgical implantation of a temporary cava filter in the prophylaxis of pulmonary artery embolism]. PMID- 11475977 TI - [Application of laparocentesis and gastrofibroscopy of pyloroduodenal tract]. PMID- 11475978 TI - [Application of videothoracoscopy in surgical treatment of thymoma and histological characteristics of the tumor]. PMID- 11475979 TI - [Application of laparoscopy in diagnosis and treatment of acute ileus]. PMID- 11475980 TI - [Pulmonary endometriosis]. PMID- 11475981 TI - [The problem of mycosis in oncology]. PMID- 11475982 TI - [Specific features of the course of infected mesenchymal hepatic hamartoma in a 2 year-old child]. PMID- 11475983 TI - [Observation of the complex biliodigestive fistula]. PMID- 11475984 TI - [Postop complications in inguinal hernia and their surgical correction]. PMID- 11475985 TI - [Diagnosis and surgical treatment of secondary gastroesophageal reflux disease]. AB - Examination and treatment was performed in 69 patients with secondary gastroesophageal reflux disease (GERD), 61 (88.4%) of whom were primarily operated on for achalasia of cardia and GERD. The choice of the operation method was based on complex examination of the patients. Due to conduction of preoperative preparation it was possible to perform the most physiological antireflux fundoplication operations in the patients. PMID- 11475986 TI - [Secondary prevention in acute myocardial infarction]. AB - Myocardial infarction is still one of the main causes of mortality and morbidity in Western countries. The advances made in the last 30 years have made it possible to reduce mortality significantly (which is currently below two digits) as well as morbidity. The subject of secondary prevention of myocardial infarction gains particular significance in this context since 10 to 15% of the patients who survive the hospital phase of myocardial infarction die during the first year following discharge and, of these deaths, half occur in the first three months. Therefore, it is necessary to make an early definition of the risk of another coronary event, that is, to make a risk stratification. This should occur throughout hospitalization and should be complete at the time of discharge, never beyond the first weeks of evolution. Bearing in mind the age, sex, coronary risk factors, ischemia persistence, the degree of left ventricular dysfunction and the presence of malignant disrhythmias, there are three risk levels: high; intermediate; and low. An overall approach to secondary prevention of infarction should take into account that, apart from the factors of such high prognostic value (Chapter II) assessed in the definition of risk groups, the measures to reduce reinfarction and sudden death (Chapter III) and the control of the risk factors for heart disease (Chapter IV) should also be considered. The principal late complications of infarction with significant prognostic influence are described in Chapter III: left ventricular dysfunction; rhythm disturbances and residual ischemia. The diagnostic criteria and therapeutic objectives are considered in each of the groups with relevance to consolidated advances according to the modern concept of evidence based medicine, according to international regulations. The grading of scientific evidence into three distinct categories (A, B and C), based on five levels of evidence classified from I to V, is presented accordingly in relation to the therapeutic proposals. Chapter III deals with a set of therapeutic interventions used in secondary prevention because they reduce reinfarction and sudden death: platelet antiaggregants; anticoagulants; Beta blockers; calcium channel blockers; antioxidants and nitrates. A concept of particular clinical significance is presented for each of these groups of drugs. The last part contains an eminently clinical overall review of the principal advances in coronary risk factor control, new therapeutic acquisitions in atherosclerotic disease with natural relevance to hypolipidemic agents and statins, which apart from controlling the plasmatic levels of cholesterol, also stabilize the atherosclerotic plaque and reduce acute coronary events significantly. Apart from dyslipidemia, the classic risk factors are: smoking; hypertension; obesity; diabetes and sedentary life. In each case, reference is made to the general measures and specific approaches, as well as the pharmacological therapy according to evidence based medicine. The recommended attitudes are pointed out. The role of cardiac rehabilitation and postmenopausal hormone replacement therapy are also discussed in the last part of these recommendations, in which the on-going controversy regarding hormone replacement therapy is pointed out in view of the results of more recent clinical trials. PMID- 11475987 TI - [Depression]. AB - The Portuguese Medical Association, through its Colleges of Specialistes, has decided to establish working groups to establish Therapeutic Guidelines in pathologies of great interest and current relevance in order to improve and rationalize health care. The group in charge of establishing the Therapeutic Recommendations in Depression, whose work I had the responsability and pleasure of coordinating during part of 1998 and 1999, is comprised of colleagues selected by the Portuguese Medical Association (Ordem dos Medicos). In the last few years, there has been rapidly increasing clinical and scientific interest in the study and treatment of depression throughout the life cycle, in different contexts and levels of medical practice. The rapid progress made in the neurosciences and psychopharmacology, new research in the field of psychotherapies and improved knowledge of relevant psychosocial aspects has not only broadened our knowledge of the aetiology and pathogenesis of depression, but also improved our criteria of diagnosis and classification. This has allowed the development of new therapeutic approaches and new drugs of proven efficacy. These Recommendations are aimed at systematizing and disseminating a consensus on interventions in depression, supported by evidence and the most recent scientific developments, in a way that will optimise therapeutic treatment. Due to some delay in the publication of these Recommendations (through no fault of the working group), and the rapid evolution of knowledge in this area, some of the contents may shortly need to be revised and updated. The writers and publishers of this document are fully aware of this and in accordance. PMID- 11475988 TI - [Psoriasis]. AB - The purpose of these Guidelines is to summarize the most relevant features of the pathogenesis, clinical presentation and treatment of psoriasis. Patient education should include the deleterious effects that some drugs, trauma, alcohol, infection and stress may have on psoriasis; the beneficial action of careful sunlight exposure should also be emphasized. Topical treatment--emollients, keratolytics, coal tar preparations, anthralin, corticosteroids, calcipotriol--is essential for the control of plaque-type psoriasis and is also an important adjuvant therapy in more severe cases; the relative strength and the potential adverse effects of topical steroids are also referred. UV therapy (phototherapy and photochemotherapy) is recommended for psoriasis with generalized plaque, guttate or palmoplantar psoriasis refractory to topical therapies. Systemic therapy--retinoids, methotrexate, cyclosporine--is limited to severe plaque psoriasis unresponsive to topical or UV therapy, erythrodermic, pustular or arthropatic psoriasis. Combination and rotational therapies are likely to reduce the risks of each individual therapy and should be encouraged. Finally, a few diagrams are included, pointing out the scientific validity of the therapies currently available to help clinicians to optimize their management of psoriasis. PMID- 11475989 TI - [Chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) includes two entities, Chronic Bronchitis (C.B.) and Emphysema (E.), assembled by their common physiopathological feature, a progressive expiratory airflow obstruction. The presence of hyperreactivity is common and partial reversibility may be observed. The coexistence of CB and E. is largely the most prevalent. Cigarette smoking is the main risk factor per se or associated with other urban or occupational air pollutants. Therefore epidemiological COPD rats follow the prevalence of smoking habits in the population. The main goal of the strategy against the disease are preventive decisions and measures, as far as smoking is concerned. FEV1 values lower than 75% of FVC and a decline of FEV1 higher than 30 ml/year indicate a high risk of COPD among smokers. The assessment of FEV1 is important for monitoring and guiding treatment and is valid for grading the severity of COPD. Once the disease is confirmed the therapeutic skills are the following: to reduce the symptoms; to prevent and treat the exacerbations; to attenuate the functional impairment, both short and long term; to achieve better performance in daily activities and quality of life; to avoid and minimise the adverse effects of medication. The increase in life expectancy and the reduction of mortality are tangible goals related to a global and multidisciplinary approach. They mainly depend on smoking cessation, reducing the bronchial obstruction, and correcting chronic hypoxemia. PMID- 11475990 TI - Restricting physician-assisted death to the terminally ill. PMID- 11475991 TI - Resolution and ambivalence. PMID- 11475992 TI - Regulating assisted reproduction. PMID- 11475993 TI - Families, patients, and physicians in medical decisionmaking: a Pakistani perspective. PMID- 11475994 TI - Groups, communities, and contested identities in genetic research. PMID- 11475995 TI - Compassionate physicians. PMID- 11475996 TI - Prayer is therapy. PMID- 11475997 TI - Research ethics: reports, scandals, calls for change. PMID- 11475998 TI - How to speak postmodern. Medicine, illness, and cultural change. PMID- 11475999 TI - [Maintenance of cellular memory by Polycomb group genes]. AB - The Polycomb-group genes (PcG) encode a group of repressors well known for their function in stably maintaining the inactive expression patterns of key developmental regulators, including homeotic genes. PcG genes are structurally and functionally conserved in Drosophila and Mammalians, and some homologues have been found in worms, yeast and plants. Their products act through different complexes and at least one of these complexes seems to induce histone deacetylation. In Drosophila, building of PcG complexes depends on both protein protein interactions and recognition near target genes of specific DNA sequences called Polycomb-group response element (PRE). Together with the counteracting trithorax-group proteins, PcG products establish a form of cellular memory by faithfully maintaining transcription states determined early in embryogenesis. Here, we discuss several aspects of PcG functions: the composition of the different complexes, the establishment and the transmission of silencing to subsequent cell generations as well as the subnuclear localisation of the PcG products. PMID- 11476000 TI - [Geographic distribution of surnames and genetic structure: the county of Ardeche at the beginning of the twentieth century]. AB - The analysis of the geographical distribution of surnames (whose transmission in the European societies is similar to that of the Y chromosome) allows to study in an exhaustive way the spatial and temporal human population structure and the influence of environmental variations on this structure. The genetic structure of the Ardeche county was analysed through a study of the surname characteristics (diversity, frequency, geographical dispersion, etc.) of individuals born between 1891 and 1915. The estimation of the intra-community average consanguinity and the inter-district genetic relationships reveals a clear differentiation between populations of the mountainous part (geographically isolated, highly inbred) and those of the Rhone valley (less isolated, weakly inbred). As predicted by the isolation by distance model, inter-population coefficients of kinship decreases as distance increases, confirming the presence of a spatial structure, characterized by neighborhood exchange and resulting from former migratory flow. PMID- 11476001 TI - Is male-infant caretaking related to paternity and/or mating activities in wild Barbary macaques (Macaca sylvanus)? AB - In species with a promiscuous mating system, the functions of male-infant caretaking remain unclear in the absence of genetic paternity tests. We tested paternal investment and hypotheses concerning reproductive tactics in wild groups of Barbary macaques, including results of genetic paternity tests. Our study revealed that male-infant caretaking was not related to the probability of paternity. In principle, males could use access to females to estimate paternity. However, we found that mating success was not related to paternity, so males could invest in infants that they had not sired, and caretaking of non-offspring was actually observed. Accordingly, males might be 'deceived' with respect to their paternal investment. In that case, one would expect a positive relation between mating success and the subsequent rate of male caretaking of infants. Such a relation is also lacking, leading to comprehensive rejection of the paternal investment hypothesis in Barbary macaques. By contrast, there was evidence that males showing infant care achieved higher mating frequencies than other males with the mothers of the relevant infants. Thus, male Barbary macaques do not show a 'mate-then-care' pattern, but they do exhibit a 'care-then-mate' pattern. PMID- 11476002 TI - The nitration of proteins in platelets. AB - Nitric oxide has many important physiological functions, but it may also form an important oxidant, peroxynitrite, as a consequence of its reaction with superoxide anions. Peroxynitrite is capable of nitrating the aromatic amino acids in proteins, particularly tyrosine. Nitrated proteins are found in tissues of a variety of diseases where inflammation occurs. However, our recent work suggests that more selective nitration of specific proteins may occur during normal physiological processes, such as platelet activation by collagen. It is not yet clear what role this may play in the normal cell biology, but there is potential to be a role in signal transduction mechanisms, possibly by influencing tyrosine phosphorylation or dephosphorylation. PMID- 11476003 TI - Haslea salstonica sp. nov. and Haslea pseudostrearia sp. nov. (Bacillariophyta), two new epibenthic diatoms from the Kingsbridge estuary, United Kingdom. AB - Two new diatom species, Haslea salstonica and Haslea pseudostrearia are described in light and electron microscopy and compared with two well-known members of Haslea. Scanning electron microscope observations confirm that the new species belong to the genus Haslea. This study extends previous observations on the genus, particularly with respect to the development of a pseudostauros. The characteristic features of the genus are discussed briefly. PMID- 11476004 TI - [The multistemmed structure of Juniperus thurifera: adaptive advantage in a severe environment?]. AB - A comparative study of radial growth and biomass between multistemmed trees with variable number of stems and single-stemmed trees was carried out to better understand determinism and organisation of multicaulis structure of a juniper species (Juniperus thurifera L.) growing in high Mediterranean mountains (High Atlas, Morocco). It appears that all the stems of the same tree have similar ages and so simultaneous development. Their mean annual radial increments show significant differences because of probable competition for water and nutrient supply and obvious physical competition for space. The multistemmed trees characterized by low number of stems have the same mean annual radial growth as single-stemmed trees and more generally all multistemmed junipers have a higher biomass. The multicaulis structure of Juniperus thurifera thus could be considered as an adaptation to severe environment, characterized not only by hard topographical, edaphic and climatic conditions, but by strong human pressure too. PMID- 11476005 TI - Spatial and temporal co-structure analyses between ichthyofauna and environment: an example in the tropics. AB - Ichthyofauna distribution and habitat characteristics of Thalassia beds in the Grand Cul-de-Sac marin lagoon in Guadeloupe were studied during a one-year survey. Environmental variables (9) were measured monthly in ten sites along with collection of fish communities. The environmental data set, analysed alone through between-within group 'principal component analysis' (PCA), exhibited a significant spatial and temporal variability. The fish data set, however, presented only a significant spatial structure, stable over the year. Given the lack of temporal variability in fish distribution, a 'between-site co-structure analysis' (BSCA) was used to compare the faunistic and environmental structures in space. The co-inertia structure was reduced to one axis representing a strong coast-reef gradient, the major common phenomena to both data sets. Environment and fish distribution allowed to distinguish sites directly under mangrove influence (characterised by high seagrasses, high concentration of chlorophyll a and high densities of zooplankton), to sites under reef influence (with short but dense seagrasses, clear water, and poor nutriments). For that purposes, the BSCA summarised efficiently what in common the fauna spatial structure and the environment spatial structure may present. PMID- 11476006 TI - Taxonomic relationships and sampling effects among Lepilemuridae and Lemuridae using a partial cytochrome b gene. AB - Partial cytochrome b sequences were used to study relationships between three Lepilemuridae species (Lepilemur dorsalis, L. septentrionalis and L. leucopus) and other Lemuridae species. L. dorsalis were subdivided into two sub-groups, according to their capture area (Nosy-Be island and Sahamalaza peninsula). Relationships deduced from phylogenetic trees as well as genetic distances lead to the classification of the Lepilemurs analysed here into separate species. These Lepilemurs form a monophyletic clade which is the sister clade of all other Lemurs used in this study. Reconstructions using randomly chosen sequences and step by step addition of sequences indicate that phylogenetic results for closely related species need to be analysed with caution, if only a small number of sequences are used to obtain them. PMID- 11476007 TI - [Urban noise pollution]. AB - Noise is responsible for cochlear and general damages. Hearing loss and tinnitus greatly depend on sound intensity and duration. Short-duration sound of sufficient intensity (gunshot or explosion) will not be described because they are not currently encountered in our normal urban environment. Sound levels of less than 75 d (A) are unlikely to cause permanent hearing loss, while sound levels of about 85 d (A) with exposures of 8 h per day will produce permanent hearing loss after many years. Popular and largely amplified music is today one of the most dangerous causes of noise induced hearing loss. The intensity of noises (airport, highway) responsible for stress and general consequences (cardiovascular) is generally lower. Individual noise sensibility depends on several factors. Strategies to prevent damage from sound exposure should include the use of individual hearing protection devices, education programs beginning with school-age children, consumer guidance, increased product noise labelling, and hearing conservation programs for occupational settings. PMID- 11476008 TI - [Emergency calls in the Samu system]. AB - The French emergency medical system is public and medical. Named Samu, it is almost entirely managed by anesthesiologists specially trained for emergency medical assistance and advanced medical support. The system comprises a reception and dispatching center for emergency calls, directly connected to police and firemen, but assuring patient confidentiality. This fixed center is managing mobile units which are Medical Intensive Care Ambulances (MICA named Smur in France). The Samu's mission consists in performing urgent individual medical assistance and also medical services for treatment of mass casualties. It is also to find adequate hospital units for the patients, to perform medical transport and to be a consultant for general practitioners in emergency. PMID- 11476009 TI - Development of an improved whitening dentifrice based upon "stain-specific soft silica" technology. PMID- 11476010 TI - Anticaries profile qualification of an improved whitening dentifrice. AB - A series of pH cycling "profile" laboratory studies was conducted to confirm the anticaries potential of an enhanced cleaning and whitening dentifrice based on a novel abrasive polishing silica and pyrophosphate tartar control source, with sodium fluoride as the anticaries ingredient. Under pH cycling remineralization conditions, the whitening dentifrice produced lesion fluoridation comparable to a conventional tartar control dentifrice containing combinations of pyrophosphate anion and a NaF fluoride ion source. Under pH cycling lesion progression conditions, the whitening dentifrice produced enamel protection against caries development, comparable to control dentifrices containing NaF alone or in combination with tartar control pyrophosphate anion. These results are consistent with clinical data supporting the effectiveness of NaF dentifrices combined with anionic tartar control inhibitors such as pyrophosphate. These results support the anticaries effectiveness of the enhanced cleaning and whitening dentifrice based upon "Stain-Specific Soft Silica." PMID- 11476011 TI - Laboratory stain removal and abrasion characteristics of a dentifrice based upon a novel silica technology. AB - These laboratory studies examined the stain removal efficacy and hard tissue abrasivity of a new dentifrice formulation--Crest Extra Whitening--based on the incorporation of elevated concentrations of a proprietary silica. Cleaning power assessments were made using a modification of the laboratory test method developed by Stookey and associates at Indiana University Oral Health Research Institute. Abrasion assessments were made using Radioactive Enamel and Radioactive Dentin Abrasivity (REA and RDA) measures. Results show that Crest Extra Whitening dentifrice produced statistically significantly improved stain removal when compared to a number of conventional dentifrices, including Crest Cavity Protection and Crest Tartar Protection, and a number of recently marketed cleaning and whitening dentifrices. Laboratory studies further demonstrated that the Crest Extra Whitening dentifrice produces dentin and enamel abrasivity similar to conventional silica dentifrices. PMID- 11476012 TI - In vitro studies of the anticalculus efficacy of an improved whitening dentifrice. AB - Crystal growth inhibition and plaque biofilm calcification laboratory studies were conducted to confirm the anticalculus potential of an enhanced cleaning and whitening dentifrice, based upon a novel enamel-safe, abrasive-polishing silica and pyrophosphate tartar control source with sodium fluoride as the anticaries ingredient. Calcium hydroxyapatite crystal growth following direct supernate treatments was significantly inhibited by the advanced whitening dentifrice. Similarly, plaque biofilm calcification was significantly inhibited by supernate treatments of the advanced whitening dentifrice. In both protocols, the advanced whitening dentifrice produced activity similar to formulations clinically proven for antitartar activity. These results provide support for the antitartar clinical activity of this new dentifrice. PMID- 11476013 TI - Removal of extrinsic stain using a tartar control whitening dentifrice: a randomized clinical trial. AB - A nine-week, double blind clinical trial was conducted to evaluate the effectiveness of a novel tartar control whitening dentifrice with a silica-based abrasive system on induced dental stain. The study model involved three weeks of stain induction followed by six weeks of unsupervised brushing to assess efficacy. To induce stain, 222 healthy adult volunteers received a dental prophylaxis, and then began a limited brushing regimen supplemented by three times daily rinsing with tea and once daily rinsing with 15 ml of 0.12% chlorhexidine. This regimen was suspended, and 187 subjects with tooth stain were entered into a six-week clinical trial where they were randomized to either a silica-based tartar control whitening dentifrice or a marketed regular dentifrice control, balancing for stain levels and smoking status. At baseline, three and six weeks, stain area and stain intensity were measured on the 8 anterior teeth using the Lobene Index. After six weeks' use, composite Lobene means were 35% lower for the whitening dentifrice compared to the regular control. In addition to the overall reductions, there were statistically significant reductions in stain area (p < 0.015) and stain intensity (p < 0.01) at both three and six weeks. The tartar control whitening dentifrice was effective in removing stain on the gingival margins and elsewhere on the body of the tooth. Safety profiles for the two test dentifrices were generally similar. After three and six weeks' use, the tartar control whitening dentifrice reduced chlorhexidine and tea stain compared to the marketed control. PMID- 11476014 TI - Effects of a tartar control whitening dentifrice on tooth shade in a population with long-standing natural stain. AB - Changing and whitening tooth color in people with long-standing tooth stain without excessive hard tissue abrasion may represent one of the more difficult challenges for whitening dentifrices. An eight-week clinical trial was conducted to evaluate change in tooth color by a silica-based, enamel-safe tartar control whitening dentifrice compared to a marketed baking soda dentifrice control. First, a screening exercise was conducted to identify individuals with long standing extrinsic dental stain. This exercise targeted adults who reported "stained teeth" and coffee/tea drinking or smoking, but who had no recent history of dental prophylaxis. Targeted subjects were examined for stain (Lobene Index) and tooth shade/color (Vita). A total of 291 adults having extrinsic stain and discolored teeth were enrolled in the study. Subjects were randomized to one of the two treatment groups, and all dentifrice use was unsupervised. Tooth color was measured at 4 and 8 weeks from shade values collected from the 8 incisors, and averages were determined from a linear ordering of the shade guide. A total of 278 evaluable subjects completed the 8-week study. Overall, the tartar control whitening dentifrice group experienced an improvement in color, differing statistically from baseline (p < 0.001) and from the marketed control (p < 0.05). Safety profiles for the two dentifrices were generally similar. Among patients with long-standing extrinsic stain, use of the tartar control whitening dentifrice resulted in superior overall tooth shade and reduced maximum or worst color compared to the marketed baking soda dentifrice control. PMID- 11476015 TI - Maintenance of tooth color after prophylaxis: comparison of three dentifrices. AB - Pellicle rapidly accumulates on tooth surfaces after prophylaxis and may acquire cosmetically unacceptable levels of stain. A three-month clinical trial was conducted to evaluate stain prevention by a new silica-based tartar control whitening dentifrice (Crest Extra Whitening) compared to marketed tartar control baking soda peroxide whitening and regular dentifrice controls. Prior to the trial, a one-month screening exercise was conducted to identify adult subjects who accumulated extrinsic tooth stain after dental prophylaxis. A total of 672 subjects were stratified based on tooth whiteness, gender and tobacco usage, then given a dental prophylaxis and randomized into one of three dentifrice treatment groups. All product use was unsupervised. Change in tooth whiteness (delta L*) was determined by comparing colorimeter measurements collected on the facial surfaces of the four central incisors at months 1 and 3 to baseline. Ninety-six percent of subjects completed the three-month study. At both one and three months, the two whitening dentifrices did not differ from baseline in terms of delta L*. In contrast, the regular control had delta L* values of -0.26 and -0.39 at one and three months, respectively, differing significantly from baseline at both time points. Each of the whitening dentifrices differed statistically from the regular control in stain accumulation (p < or = 0.001) at one and three months, but were not different from each other. In general, all three test dentifrices were well tolerated. These data demonstrate the effectiveness of the new tartar control whitening dentifrice in preventing stain accumulation after dental prophylaxis compared to the marketed regular dentifrice control. PMID- 11476016 TI - [Seeking tuberculosis care in Chiapas, Mexico]. AB - OBJECTIVE: To analyze the process of seeking tuberculosis care and this process's impact on treatment adherence in the Mexican state of Chiapas, given that the two primary factors in successful tuberculosis (TB) control programs are early diagnosis and adherence to TB treatment. METHODS: We conducted a qualitative study using group interviews with 11 groups of patients in three of the nine socioeconomic regions of the state of the Chiapas (Altos, Centro, and Fronteriza). RESULTS: The patients applied a variety of approaches in seeking care. The patients reported considerable delays in diagnosis due to problems that the patients themselves had and because of shortcomings in the care they received from the formal health care system. The treatment options that they followed were the result of their perceptions of the causes of TB and of the variety of traditional medical practices accepted in their communities. CONCLUSIONS: The lack of knowledge about TB encourages people to consider various alternatives for their care. Tuberculosis control in Chiapas requires an optimal utilization of the health services that exist in the state as well as a program of health education. TB control in Chiapas must take into account the social, cultural, and economic reality of the population. PMID- 11476017 TI - [Iron deficiency anemia and nutrition in the second year of life in Rio de Janeiro, Brazil]. AB - OBJECTIVE: To assess the influence of feeding practices on the prevalence of anemia among breast-fed children, based on the nutritional profile of children receiving care at an outpatient pediatric clinic in the city of Rio de Janeiro, Brazil. METHODS: This was a cross-sectional study with 288 children between 12 and 18 months of age who were seen at the outpatient clinic between January and June 1993. The children were assessed in terms of the presence of iron-deficiency anemia. In addition, two questionnaires were done with the child's mother or the other person who had brought the child to the clinic: a 24-hour dietary recall and a record of the frequency of the consumption of iron-rich foods. RESULTS: Of the 288 children, 144 of them had signs of anemia (hemoglobin < 11 g/dL); of these 144, 38 of them had severe anemia (hemoglobin < 9.5 g/dL). We found low levels of bioavailable iron in the foods consumed, limited consumption of meat, and vitamin C consumption separate from meals. We found a significant association between the prevalence of severe anemia and inadequate iron intake (relative risk = 2.28; 95% confidence interval = 1.12 to 4.66; P = 0.02). The intake of bioavailable iron was higher in the group without anemia (P = 0.04). CONCLUSIONS: Individuals caring for breast-fed children should be educated concerning the nutritional contents of complementary foods so as to increase the bioavailability of iron in the children's diets. One way to achieve this objective might be through "Integrated Management of Childhood Illnesses," a strategy endorsed by a number of international organizations as a way to reduce child mortality and morbidity in developing countries. PMID- 11476018 TI - [Aluminum hydroxide hydrogel to remove arsenic from water]. AB - OBJECTIVE: To describe the making of and the preliminary results from an aluminum hydroxide hydrogel that, when added directly to water, can totally eliminate arsenic, regardless of the nature of the water and the oxidation state of the metalloid. METHODS: The raw materials used to make the aluminum hydroxide hydrogel were: hydrated aluminum sulfate (which is used to make water safe for drinking), calcium hypochlorite in powder form, ammonium hydroxide, and distilled water (when done at the laboratory scale), and water from inverse osmosis of the same or better quality than the distilled water (when done at a pilot scale). Quality control for the finished product consisted of determining its ability to adsorb arsenic and performing bacteriological tests to demonstrate its sterility. The product was used with water samples to which arsenic had been added in the laboratory and with water samples from the province of Tucuman, Argentina, with naturally occurring arsenic. To analyze the arsenic in the water the colorimetric silver diethyldithiocarbamate method was used. RESULTS: Using the hydrogel greatly reduced the amount of arsenic. Treating both the natural and laboratory produced arsenical waters yielded arsenic concentrations under the limit (0.01 parts per million) that was detectable with the analytical method used. The bacteriological tests of the finished product indicated there were no viable bacteria. CONCLUSIONS: Using the aluminum hydroxide hydrogel produced the desired reduction in the arsenic concentration in the water. This method for removing arsenic is inexpensive and easy to use in scattered rural populations in areas having high arsenic levels and lacking drinkable water, as well as an adequate sanitary and electric-power infrastructure. PMID- 11476019 TI - Evolution of the health sector and tuberculosis control in Brazil. AB - OBJECTIVE: To summarize the epidemiological situation of tuberculosis (TB) in Brazil, especially as it relates to the evolution of the health sector in recent decades, the process of health sector reform, and current proposals of the Brazilian Ministry of Health. METHODS: A review was conducted of data from the Ministry of Health of Brazil on tuberculosis in the country over the last 20 years, as well as of the history of changes in the health sector. RESULTS: There have been major changes in the epidemiological situation of TB and also in the structure of the health system in Brazil. CONCLUSIONS: The overall prospects are promising for Brazil's National Plan for Tuberculosis Control. PMID- 11476020 TI - [Games as an alternative for teaching basic health concepts]. AB - OBJECTIVE: To determine, for the teaching of basic health concepts to school-age children, the effectiveness of an educational strategy based on traditional children's games. METHODS: Intervention study carried out in the city of Durango, Mexico, in June 2000 with 300 children from 9 to 11 years old. The children were randomly divided into two groups. The children in Group A used a modified version of a Mexican popular game called Serpientes y Escaleras (Snakes and Ladders) that included messages on basic health concepts; the children in Group B made up the control group and did not play the modified game. RESULTS: At baseline there were no significant differences between the two groups in terms of age, grade level, or their scores on a knowledge test of basic health concepts. After the educational intervention, the health concepts test scores, out of a maximum possible of 10, were 9.3 +/- 0.8 for Group A and 7.5 +/- 1.1 for Group B (P < 0.001). CONCLUSIONS: Using games that include health and hygiene messages can be an alternative for teaching basic health concepts. PMID- 11476021 TI - [Reference values for lead in blood in urban population in southern Brazil]. AB - OBJECTIVE: To describe the reference values for lead in blood in an urban population in the city of Londrina, in the state of Parana, Brazil. MATERIALS AND METHODS: The reference population was composed of 520 adult volunteers who were assessed from November 1994 to December 1996. Exclusion criteria were: occupational exposure to lead, exposure through personal habits or practices, smoking more than 10 cigarettes per day, and living near industrial plants or other places that use lead in their production processes. Also excluded were individuals with abnormal clinical or laboratory results or with chronic diseases or cardiovascular disorders. Lead blood levels were determined using air acetylene flame atomic absorption spectrophotometry. The detectable limit was 1.23 micrograms/dL. After the analyses of lead in blood, the following values were determined: minimum value, first quartile, median, third quartile, and maximum value; geometric mean; 95% confidence interval; experimental interval; and reference value. RESULTS: The reference values for lead in blood ranged from 1.20 micrograms/dL to 13.72 micrograms/dL. The geometric mean was 5.5 micrograms/dL. CONCLUSIONS: In general, the values found in this study are lower than those that have been reported for other countries. Additional data should be gathered from Brazilian populations living in more-industrialized areas. PMID- 11476022 TI - Brazil fights for affordable drugs against HIV/AIDS. PMID- 11476023 TI - [Detection, evaluation, and treatment of high blood cholesterol in adults]. AB - This report is based on the Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults, which was recently issued by the National Institutes of Health of the United States of America. Also known as the Adult Treatment Panel (ATP) III, this new report updates two earlier such reports on high cholesterol. While continuing to concentrate on treating patients with coronary heart disease, the new report advocates more intensive treatment in order to reduce low-density lipoprotein (LDL) cholesterol in specific groups of individuals, pays special attention to primary prevention among patients with multiple risk factors, and recognizes as a secondary prevention concern a cluster of heart disease risk factors known as "the metabolic syndrome." Other issues that the ATP III report covers include therapeutic lifestyle changes to reduce LDL, LDL-lowering drug therapy, and the management of specific dyslipidemias. PMID- 11476024 TI - [Recommendations on the use of antiretroviral agents in pregnant women infected with HIV-1]. AB - In February 1994, a clinical trial by the Pediatrics AIDS Clinical Trials Group (PACTG 076) demonstrated, for the first time, that it was possible to reduce the risk of vertical transmission of human immunodeficiency virus type 1 (HIV-1) by nearly 70% by means of a triple therapeutic regimen: 1) oral zidovudine (ZDV) beginning anytime between the 14th and 34th week of gestation until the end of pregnancy; 2) intravenous ZDV during pregnancy, and 3) administration of oral ZDV to neonates during the first 6 weeks of life. Later, epidemiologic studies performed in the United States of America and France showed that this regimen drastically reduced perinatal transmission in clinical practice. Since then, important strides have been made, not only in terms of treatment (new drugs and highly effective therapeutic regimens) and diagnosis (tests that can measure the viral load), but also in terms of an increased understanding of the pathogenesis of perinatal transmission of HIV-1. This report contains: a) a review of special considerations to be kept in mind when administering antiretroviral agents to pregnant women; b) a current overview of the results of clinical and epidemiologic trials dealing with the prevention of perinatal transmission of HIV 1; c) a look at the use of tests for measuring HIV-1 RNA (viral load) during pregnancy; d) the most recent recommendations on the use of antiretroviral chemoprophylaxis and elective cesarean section for reducing perinatal viral transmission. These recommendations apply to the United States, and other countries may prefer to use different approaches. PMID- 11476025 TI - Doc Holliday, D.D.S. 150th anniversary. PMID- 11476026 TI - A comparison of the visibility of caries on Kodak F-speed (insight) and D-speed (Ultra-speed) films. PMID- 11476027 TI - Characteristics, satisfaction, and perceptions of patients receiving ambulatory healthcare from osteopathic physicians: a comparative national survey. AB - A national telephone survey was conducted in 1998 using random-digit dialing and the first Osteopathic Survey of Healthcare in America (OSTEOSURV-I) instrument to determine patients' satisfaction with their healthcare, as well as their perceptions of osteopathic medicine. Of the 1106 respondents, 243 (22.0%) had received medical care from an osteopathic physician, and another 307 (27.8%) claimed to be aware of osteopathic physicians. Patients of osteopathic physicians reported the highest levels of satisfaction in 8 of the 11 elements studied when compared with patients of allopathic physicians, chiropractors, and nonphysician clinicians other than chiropractors. Respondents perceived osteopathic manipulative treatment (OMT) to be beneficial for musculoskeletal disorders (P < .001). In addition, respondents perceived that healthcare services provided by osteopathic physicians were similar to those provided by allopathic physicians (P < .001), but not to those provided by chiropractors (P = .01). A total of 97.9% of current patients of osteopathic physicians agreed with the statement that osteopathic physicians practiced in their local community, compared with 80.6% of former patients of osteopathic physicians and 67.8% of patients who had never visited osteopathic physicians (P < .001). In general, the most favorable perceptions of osteopathic medicine were reported by current patients of osteopathic physicians, followed by former patients of such physicians. The least favorable perceptions came from patients who had never been patients of osteopathic physicians. The perception that OMT should be covered by health insurance was significantly associated with the use of osteopathic physicians (odds ratio, 3.2; 95% confidence interval, 1.5 to 6.7, among patients who had ever been to an osteopathic physician). The results of our survey suggest that greater access to osteopathic services, including OMT, is desirable and that promotional efforts aimed at encouraging the use of osteopathic medical services among the general population are warranted. PMID- 11476028 TI - Weaning from mechanical ventilation: an update. AB - Many patients admitted to the intensive care unit have respiratory failure and thus require mechanical ventilation. Weaning patients from mechanical ventilation after their primary disease process has been treated can be difficult in approximately 30% of patients. Inadequacies in pulmonary gas exchange and in the performance of the respiratory muscle pump are the most common causes for failure to wean. Assessing whether a patient can be weaned from mechanical ventilation involves two major factors: (1) examining the patient for evidence of an increase in the work of breathing, and (2) measuring spontaneous breathing variables. Although different modalities have been used in weaning patients from mechanical ventilation, none has been shown to be more successful than repeated trials of spontaneous breathing. PMID- 11476029 TI - Evaluation of aminotransferase elevations in a bodybuilder using anabolic steroids: hepatitis or rhabdomyolysis? AB - The use of anabolic steroids among competitive athletes, particularly bodybuilders, is widespread. Numerous reports have noted "hepatic" dysfunction secondary to anabolic steroid use based on elevated serum aminotransferase levels. The authors' objective was to assess whether primary care physicians accurately distinguish between anabolic steroid-induced hepatotoxicity and serum aminotransferase elevations that are secondary to acute rhabdomyolysis resulting from intense resistance training. Surveys were sent to physicians listed as practicing family medicine or sports medicine in the yellow pages of seven metropolitan areas. Physicians were asked to provide a differential diagnosis for a 28-year-old, anabolic steroid-using male bodybuilder with an abnormal serum chemistry profile. The blood chemistries showed elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT), and creatine kinase (CK) levels, and normal gamma-glutamyltransferase (GGT) levels. In the physician survey (n = 84 responses), 56% failed to mention muscle damage or muscle disease as a potential diagnosis, despite the markedly elevated CK level of the patient. Sixty-three percent indicated liver disease as their primary diagnosis despite normal GGT levels. Prior reports of anabolic steroid-induced hepatotoxicity that were based on aminotransferase elevations may have overstated the role of anabolic steroids. Correspondingly, the medical community may have been led to emphasize anabolic steroid-induced hepatotoxicity and disregard muscle damage when interpreting elevated aminotransferase levels. Therefore, when evaluating enzyme elevations in patients who use anabolic steroids, physicians should consider the CK and GGT levels as essential elements in distinguishing muscle damage from liver damage. PMID- 11476030 TI - Student perceptions of osteopathic manipulative treatment after completing a manipulative medicine rotation. AB - Recent studies have demonstrated a decline in the use of osteopathic manipulative treatment (OMT) by osteopathic physicians, reflecting a trend that may begin in medical school. The authors used a questionnaire to examine the teaching and use of OMT in five rotations and the perceptions of 86 graduating osteopathic medical students of their experiences following their core manipulative medicine rotation. Most students indicated that they applied osteopathic principles sometimes (39.5%) or often (29.1%) during rotations. Forty-three percent of students rated their ability to apply osteopathic principles as average. The number of students who indicated that they rarely used OMT during their rotations was 31 (36.0%) for internal medicine, 21 (24.4%) for surgery, 23 (26.7%) for pediatrics, and 24 (27.9%) for obstetrics/gynecology. When asked why OMT was not used during a rotation, 47.2% of respondents cited time constraints, and 21.7% stated that their attending physicians discouraged the use of OMT. These results demonstrate a distinction between students' perceived level of osteopathic principles and skills and their application during clinical rotations. PMID- 11476031 TI - Single-tooth implant-supported restorations. Planning for an aesthetic and functional solution. AB - The single-tooth implant has found widespread support within the field of implant dentistry, with numerous studies reporting high success rates that surpass those recorded for splinted implant bridgework. Improvements in implant design, range of prosthetic components and restorative materials have made it possible to achieve optimal functional and cosmetic results. Coupled with an appropriate approach to treatment planning and patient assessment, use of the single-tooth implant may be considered a highly predictable treatment solution. PMID- 11476032 TI - Microscopic management of endodontic procedural errors: perforation repair. AB - Iatrogenic root perforation is a procedural error that complicates treatment and can lead to loss of the tooth. A number of new instruments and techniques have been introduced into endodontic practice over the last few years to address this problem. The recent addition of the operating microscope to improve the field of vision has provided the operator with the benefit of magnification and enhanced illumination. Although there are few studies evaluating objectively the use of the operating microscope, its clinical application demonstrates the possibility of achieving more precise endodontic practice. The use of the operating microscope in the non-surgical management of a mid-root perforation is reported in this paper. PMID- 11476033 TI - Viral infections of the oral mucosa and perioral region. AB - Viral infections of the oral mucosa and perioral region are commonly encountered in the practice of dentistry. The accurate and timely diagnosis of such infections, coupled with the institution of appropriate treatment, can often permit quick resolution of the condition with minimal discomfort and anxiety for the patient (and carers) and prevent the spread of infection to others, especially immunocompromised individuals. This article outlines the clinical presentation and appropriate management of common viral infections of the oral mucosa and perioral region. PMID- 11476034 TI - Focus groups--what are they? AB - The use of focus groups has become common in many spheres of research, including dental research. This article provides an update on the history of focus groups, defines focus groups in the context of academic research and highlights their possible uses. Practical advice is provided on the conduct of focus groups and ethical considerations are discussed. PMID- 11476035 TI - Occlusal splints and temporomandibular disorders: why, when, how? AB - Occlusal splints are one form of treatment in the management of patients with a temporomandibular disorder. Appliances are often used in conjunction with other forms of treatment such as physiotherapy or medication. A variety of splints is described in the literature and the dentist must ensure that the splint prescribed is of a design that has a proven success rate for the specific diagnosis. General principles that apply to the provision of all splints are outlined in this paper. PMID- 11476036 TI - An update on the physiology of the dentine-pulp complex. AB - The properties of dentine and pulp are closely related and from a functional standpoint these tissues are often referred to as the dentine-pulp complex. In recent years there has been considerable progress in our understanding of the physiology of the dentine-pulp complex. This review describes some of the advances that are relevant to clinical dentistry, including dentine permeability, dentine bonding, dentine sensitivity, the properties of pulpal nerves and blood vessels and the reactions of the dentine-pulp complex to operative dental procedures. PMID- 11476037 TI - Dens evaginatus on the labial surface of a central incisor: a case report. AB - A 7-year-old girl in the mixed dentition phase presented with abnormal morphology of the upper right central incisor. The diagnosis was a dens evaginatus, an anomaly of odontogenesis of unknown aetiology. This article describes the management of this anomaly and discusses the importance of maintaining space in the developing dentition. The aetiology and natural history of dens evaginatus are also considered. PMID- 11476038 TI - [Pharmacy counselling and antiretroviral therapy compliance: a patient survey]. AB - All patients must be in position to take advantage of advances in pharmaceutical therapy as well as information on their treatment when medicine-taking scheme are complex and/or undesirable effects are expected. That is why in 1996 we set up a pharmacy consultation carried out by staff specially trained for it. The two objectives of this study are, on the one hand, to assess the patient's compliance and also their knowledge on the medical prescription with the help of a questionnaire and on the other hand, to check the implementation by the staff of our procedures (handing over medicine-taking scheme whenever there has been a change in the patient's treatment). The answers to the questionnaire have shown 73% of the patients totally complied with the treatment, the most commonly-used means not to forget the taking of the treatment is the pill box, the necessity of taking the medicines regularly is most frequently considered as something preventing the patient's good compliance (37.5%), and 81% of the patients think they are in good or excellent spirits. However, 63% of the patients declare they have not been given a medicine-taking scheme. The result must be weighted for the patients may not remember things with accuracy. As a conclusion, our survey has shown treatment compliance is close to that described in the literature on the subject and it encourages us to change the course of the consultation. PMID- 11476039 TI - [Depleted uranium: sources, exposure and health effects]. PMID- 11476040 TI - [Pneumonia in calves: characterization of the bacterial spectrum and the resistance patterns to antimicrobial drugs]. AB - The population under study included young calves with pneumonia (group A, n = 13) and their controls (group B, n = 9), as well as older calves from which the lungs with (group C, n = 90) or without (group D, n = 10) lesions were collected after slaughter. Arcanobacterium pyogenes was the organism most commonly isolated from calves in group A (46%), followed by Haemophilus somnus (23%), Mannheimia haemolytica (15%), Streptococcus suis and Pasteurella multocida (7.7% each). Only S. suis (22%) and P. multocida (11%) were found in group B. P. multocida was isolated from 32% group C calves, H. somnus from 11%, A. pyogenes from 7.8%, M. haemolytica from 2.2% and S. suis from 1.1%. No specific pathogens were isolated in group D. Prevalence of Mycoplasma bovis infection was 69% in group A and 37% in group C. Ninety-eight strains were tested for resistAnce to antibiotics. Resistance to penicillin and ampicillin was present only in M. haemolytica (46%). High percentages of resistant strains were observed for streptomycin (48-100%), tetracycline (15-43%), sulfonamides alone (14-100%) or in combination with trimethoprim (0-100%). Therapeutic approaches to bacterial calf pneumonia in the area under study should be modified according to the isolated bacterial population, the observed antimicrobial resistances and the growing importance of Mycoplasma bovis. PMID- 11476041 TI - [Implantation of synthetic mesh for the closing of abdominal wall ruptures in the ventral flank of cows: a retrospective study of 16 cases]. AB - In this paper the technique and long-term results for abdominal wall ruptures in the ventral flank are described in 16 cows that underwent surgery between January 1990 and October 1999. Most injuries were caused by a horn of another cow. In three cases the rupture was repaired longer than 6 weeks after traumatic injury. The other defects were treated surgically 6.4 (0-25) days after they occurred. The muscle defects were closed layer by layer with simple continuous sutures (polyglactin 910, 6 metric) under general anesthesia in lateral recumbency and the sutured defect was reinforced with a synthetic mesh (polyester or polyglactin 910) fixed to the outside of the external oblique abdominal muscle. The most frequent postoperative complication was subcutaneous seroma. It was treated successfully by incision and drainage. All patients were released 11.6 +/- 5.5 (6 23) days after surgery. A telephone survey 53 +/- 26 (7-106) months after surgery revealed that the patients had an average survival time of 30 (2-104) months, had born one to seven calves without any complications and that only one cow had had an unsatisfactory milk yield. One cow had to be slaughtered 2 months after surgery because of a relapse. Synthetic mesh was used successfully to close the defect in 15 animals. The functional as well as the cosmetic result of the described operation was good to excellent. PMID- 11476042 TI - [Perineal hernia in dogs--colopexy, vasopexy, cystopexy and castration as elective therapies in 32 dogs]. AB - In 32 male dogs colopexy, vasopexy, cystopexy and castration was performed for the treatment of perineal hernia. Recurrence rate in this study is 22%, what is comparable to other studies using different methods. The degree of severity and the number of complications is lower with this technique than with others. Enlargement of the prostate was evident in 59% and bladder retroflexion in 22% of the dogs. A celiotomy allows to recognize, assess, reduce and fix displaced organs which is not possible by using other methods. The aim is to regain the tubular structure of the ampulla recti and to fix prostate and bladder cranioventrally to the pelvic entrance. The castration performed at the same time causes the prostate gland to atrophy within 2-3 weeks, what makes the pelvic entrance even wider and the dogs return to normal defecation. PMID- 11476043 TI - [Hepatic changes in meat inspection of a wild boar]. PMID- 11476044 TI - [Duplex ultrasound of the arteries and veins. I: Basic principles and extracranial arteries supplying the brain]. PMID- 11476045 TI - [Renal failure, thrombocytopenia and hemolytic anemia in a 36-year-old patient]. PMID- 11476046 TI - [Recurrent hypocalcemic tetany, generalized seizures and increased bronchopulmonary infections in an 18-year-old patient]. PMID- 11476047 TI - [Estrogen substitution after breast carcinoma?]. PMID- 11476048 TI - [Drug therapy of obstructive lung diseases]. PMID- 11476049 TI - [Inhalational corticosteroids in COPD. THe ISOLDE Study (Inhaled Steroids in Obstructive Lung Disease in Europe)]. PMID- 11476050 TI - [Intensive care--an interdisciplinary responsibility. Report of the 3rd "Interdisciplinary Intensive Care Medicine" Seminar Congress 4-9 March 1001 in Garmisch-Partenkirchen]. PMID- 11476051 TI - [Coma from the internal medicine and neurologic viewpoint]. PMID- 11476052 TI - [Cerebral ischemic accidents. Etiology, therapy and prevention]. PMID- 11476053 TI - [Epileptic seizures in advanced age from the neurologic and internal medicine viewpoint]. PMID- 11476054 TI - [Infectious inflammatory diseases of the central nervous system from the neurological and internal medicine viewpoint]. PMID- 11476055 TI - [Paraneoplastic diseases of the central nervous system from the neurologic and internal medicine viewpoint]. PMID- 11476056 TI - Death out West: the link to COPD. PMID- 11476057 TI - Moving beyond MTBE. PMID- 11476058 TI - New drugs engineered to fight asthma. PMID- 11476059 TI - Cisapride use in preterms. PMID- 11476062 TI - Anesthesia administration to guinea pigs: response to isoflurane anesthesia in guinea pigs [39(2):43], M. C. Ross et al. PMID- 11476063 TI - Tolerance of haemodynamic changes during beating heart coronary surgery. PMID- 11476064 TI - Helicobacter pylori and cardiac surgery. PMID- 11476065 TI - PTEN mutations are uncommon in Proteus syndrome. PMID- 11476066 TI - Limited contribution of interchromosomal gene conversion to NF1 gene mutation. PMID- 11476067 TI - A clinical study of patients with multiple isolated neurofibromas. PMID- 11476068 TI - B cell immunodeficiency, distal limb abnormalities, and urogenital malformations in a three generation family: a novel autosomal dominant syndrome? PMID- 11476069 TI - Chromosome 2 interstitial deletion (del(2)(q14.1q21)) associated with connective tissue laxity and an attention deficit disorder. PMID- 11476070 TI - Antibiotic use. PMID- 11476071 TI - How to build a testis. PMID- 11476072 TI - Applications for GnRH antagonists. PMID- 11476074 TI - Rational development of peptide vaccination in clinical oncology. PMID- 11476075 TI - Concerning Bembenek A, Schlag PM: Sentinel lymphadenectomy--ready for clinical routine. Onkologie 2000:23:408-409. PMID- 11476076 TI - Significance of chromosomal aberrations in CLL. PMID- 11476077 TI - Exposures, mutations and the history of causality. PMID- 11476078 TI - Glutaraldehyde induced asthma in endoscopy nursing staff. PMID- 11476079 TI - Ulcerative colitis is more strongly linked to chromosome 12 than Crohn's disease. PMID- 11476080 TI - Intestinal permeability: the cellobiose/mannitol test. PMID- 11476081 TI - Evaluation of the role of CFTR in alcohol related pancreatic disease. PMID- 11476082 TI - Alcohol, obesity, and TNF-alpha. PMID- 11476083 TI - Expandable metal stent placement for gastric outlet obstruction. PMID- 11476084 TI - Is routine topical anesthesia or conscious sedation necessary in upper endoscopy, especially in patients with liver cirrhosis? PMID- 11476085 TI - Serious complications with neck manipulation and informed consent. PMID- 11476086 TI - The pandas' habitat at Wolong Nature Reserve. PMID- 11476087 TI - The pandas' habitat at Wolong Nature Reserve. PMID- 11476088 TI - Social vs natural science? PMID- 11476089 TI - UNSCEAR on the health effects from Chornobyl. PMID- 11476090 TI - Captured by art. PMID- 11476091 TI - Do frequency representations eliminate conjunction effects? An exercise in adversarial collaboration. PMID- 11476092 TI - Higher-education administrators: when the future does not make a difference. AB - The relationships between administrative actions and their long-term consequences were investigated in 14 people beginning training to become high-level college and university administrators, 19 people who had just finished that training, and 44 provosts (chief academic officers) at colleges and universities in the northeastern United States. The experienced administrators (those who had completed the training plus the provosts) were significantly more likely than the trainees to mention long-term consequences when describing their past and possible future administrative actions. However, in hypothetical choice situations, the experienced administrators were also significantly more likely than the trainees to choose smaller amounts offunds available immediately for their units versus larger amounts of promised future funds. With experience, administrators may both become more aware of their actions' long-term consequences and learn that they are unlikely to receive promised future funds. The contingencies in effect for higher-education administrators may lead them to make choices that do not result in their institutions meeting the highest standards. PMID- 11476093 TI - Linguistically mediated visual search. AB - During an individual's normal interaction with the environment and other humans, visual and linguistic signals often coincide and can be integrated very quickly. This has been clearly demonstrated in recent eye tracking studies showing that visual perception constrains on-line comprehension of spoken language. In a modified visual search task, we found the inverse, that real-time language comprehension can also constrain visual perception. In standard visual search tasks, the number of distractors in the display strongly affects search time for a target defined by a conjunction of features, but not for a target defined by a single feature. However we found that when a conjunction target was identified by a spoken instruction presented concurrently with the visual display, the incremental processing of spoken language allowed the search process to proceed in a manner considerably less affected by the number of distractors. These results suggest that perceptual systems specializedfor language and for vision interact more fluidly than previously thought. PMID- 11476094 TI - Visual search has memory. AB - By monitoring subjects' eye movements during a visual search task, we examined the possibility that the mechanism responsible for guiding attention during visual search has no memory for which locations have already been examined. Subjects did reexamine some items during their search, but the pattern of revisitations did notfit the predictions of the memory less search model. In addition, a large proportion of the refixations were directed at the target, suggesting that the revisitations were due to subjects' remembering which items had not been adequately identified. We also examined the patterns of fixations and compared them with the predictions of a memoryless search model Subjects' fixation patterns showed an increasing hazard function, whereas the memoryless model predicts a flat function. Lastly, we found no evidence suggesting that fixations were guided by amnesic covert scans that scouted the environment for new items during fixations. Results do not support the claims of the memorvless search model, and instead suggest that visual search does have memory. PMID- 11476095 TI - Visual implicit memory in the left hemisphere: evidence from patients with callosotomies and right occipital lobe lesions. AB - Identification of visually presented objects and words is facilitated by implicit memory for past visual experiences with those items. Several behavioral and neuroimaging studies suggest that this form of memory is dependent on perceptual processes localized in the right occipital lobe. We tested this claim by examining implicit memory in patients with extensive right occipital lobe lesions, using lexical-decision mirror-reading, picture-fragment, and word fragment-completion tests, and found that these patients exhibited normal levels of priming. We also examined implicit memory in patients with complete callosotomies, using standard and divided-visual-field word-fragment-completion procedures, and found that the isolated left hemisphere exhibited normal priming effects. The results indicate that the right occipital lobe does not play a necessary role in visual implicit memory, and that the isolated left hemisphere can support normal levels of visual priming in a variety of tasks. PMID- 11476096 TI - Genetic influence on the expression of hand preferences in chimpanzees (Pan troglodytes): evidence in support of the right-shift theory and developmental instability. AB - Genetic mechanisms have been proposed to explain the pervasive representation of right-handedness in humans, whereas random, nongenetic factors have been posited to explain the lack of population-level right-handedness in nonhuman primates. We report evidence that hand preferences in chimpanzees are heritable, even among related individuals raised in different environments. Furthermore, we report that the degree of heritability is modified by factors associated with developmental instability, notably, offspring parity. The data are interpreted to reconcile both genetic models for handedness and hypotheses suggesting that developmental instability influences variation in handedness. PMID- 11476097 TI - Trace and delay eyeblink conditioning: contrasting phenomena of declarative and nondeclarative memory. AB - We tested the proposal that trace and delay eyeblink conditioning aref fundamentally different kinds of learning. Strings of one, two, three, or four trials with the conditioned stimulus (CS) alone and strings of one, two, three, or four trials with paired presentations of both the CS and the unconditioned stimulus (US) occurred in such a way that the probability of a US was independent of string length. Before each trial, participants predicted the likelihood of the US on the next trial. During both delay (n = 20) and trace (n = 18) conditioning, participants exhibited high expectation of the US following strings of CS-alone trials and low expectation of the US following strings of CS-US trials--a phenomenon known as the gambler's fallacy. During delay conditioning, conditioned responses (CRs) were not influenced by expectancy but by the associative strength of the CS and US. Thus, CR probability was high following a string of CS-US trials and low following a string of CS-alone trials. The results for trace conditioning were opposite. CR probability was high when expectancy of the US was high and low when expectancy of the US was low: The results show that trace and delay eyeblink conditioning are fundamentally different phenomena. We consider how the findings can be understood in terms of the declarative and nondeclarative memory systems that support eyeblink classical conditioning. PMID- 11476098 TI - Men and women at promise for scientific excellence: similarity not dissimilarity. AB - U.S. math-science graduate students possessing world-class talent (368 males, 346 females) were assessed on psychological attributes and personal experiences in order to examine how their talents emerged and developed. Comparisons were made, using similar assessments, with mathematically talented students (528 males, 228 females) identified around age 13 and tracked into adulthood by the Study of Mathematically Precocious Youth (SMPY). Well before college, both samples were academically distinguished: however, the graduate students could be identified during adolescence as a subset of mathematically talented youths based on their nonintellectual attributes. Their profiles corresponded to what earlier psychological studies found to characterize distinguished (and exclusively male) scientists: exceptional quantitative reasoning abilities, relatively stronger quantitative than verbal reasoning ability, salient scientific interests and values, and finally, persistence in seeking out opportunities to study scientific topics and develop scientific skills. On these attributes, sex differences were minimal for the graduate students (but notfor the SMPY comparison groups). Developing exceptional scientific expertise apparently requires special educational experiences, but these necessary experiences are similar for the two sexes. PMID- 11476099 TI - Absolute identification with simple and complex stimuli. AB - It is well known that people can perfectly identify only a handful of distinct unidimensional stimuli, such as line lengths, but can identify thousands of complex stimuli, such as letters and words. This result is consistent with capacity limits in identifying unidimensional stimuli but not complex stimuli. The experiments reported here tested this theoretical dissociation using Luce's (1963) Similarity Choice Model to measure the psychological distance between stimuli in line-length-identification and letter-identification tasks. The psychological distance between line-length stimuli decreased with the number of to-be-identified stimuli; this result is concordant with capacity limits in unidimensional absolute identification. Surprisingly, the opposite result held in letter identification. Psychological distance between letters increased with an increased number of to-be-identified stimuli. This result indicates an opposite type of processing deficit: People process letters more efficiently with more choices. PMID- 11476100 TI - Children creating language: how Nicaraguan sign language acquired a spatial grammar. AB - It has long been postulated that language is not purely learned, but arises from an interaction between environmental exposure and innate abilities. The innate component becomes more evident in rare situations in which the environment is markedly impoverished. The present study investigated the language production of a generation of deaf Nicaraguans who had not been exposed to a developed language. We examined the changing use of early linguistic structures (specifically, spatial modulations) in a sign language that has emerged since the Nicaraguan group first came together: In tinder two decades, sequential cohorts of learners systematized the grammar of this new sign language. We examined whether the systematicity being added to the language stems from children or adults: our results indicate that such changes originate in children aged 10 and younger Thus, sequential cohorts of interacting young children collectively: possess the capacity not only to learn, but also to create, language. PMID- 11476101 TI - Evidence for two components of object-based selection. AB - A wealth of research has shown that observers can bias visual processing toward specific locations, but the role of object-based selection is less clear In support of object-based selection, previous research has shown that when two objects are presented simultaneously, observers are better at reporting two attributes from one of the objects than one attribute from each object. However there has been controversy over whether this effect is best explained by object based selection or spatial selection. Our work suggests that there are two separate components of selection in this task: (Ca) a spatial component that is observed when the relevant targets are cued for observers before the onset of the stimulus display and (b) an object-based component that can still be observed when the first component has been eliminated. The latter effect replicates the initial evidence in favor of object-based selection, and can be demonstrated even when the relevant targets are cued after the offset of the target stimuli. PMID- 11476102 TI - All together now: when dissociations between knowledge and action disappear. AB - Why do people sometimes seem to know things but fail to act appropriately on the basis of this knowledge? Such dissociations between knowledge and action often occur in infants and children, and in adults following brain damage. These dissociations have supported inferences about the organization of cognitive processes (e.g., separable knowledge and action systems) and their development (e.g., knowledge systems develop before action systems). The current study tested the basis for knowledge-action dissociations in a card-sorting task in which children typically correctly answer questions about sorting rules while sorting cards incorrectly. When questions and sorting measures were more closely equated for the amount of conflict that needed to be resolved for a correct response, children showed no systematic dissociation between knowledge and action. The results challenge standard interpretations of knowledge-action dissociations and support an alternative account based on graded knowledge representations. PMID- 11476103 TI - Detection of glass patterns by pigeons and humans: implications for differences in higher-level processing. AB - Glass patterns have been used to examine mechanisms underlying form perception. The current investigation compared detection of Glass patterns by pigeons and humans and provides evidence for substantial species differences in global form perception. Subjects were required to discriminate, on a simultaneous display, a random dot pattern from a Glass pattern. Four different randomly presented Glass patterns were used (concentric, radial, parallel-vertical, and parallel horizontal). Detection thresholds were measured by degrading the Glass patterns through the addition of random noise. For both humans and pigeons, discrimination decreased systematically with the addition of noise. Humans showed detection differences among the four patterns, with lowest thresholds to radial and concentric patterns and highest thresholds to the parallel-horiZontal pattern. Pigeons did not show a detection difference across the four patterns. Implications for differences in neural processing of complex forms are discussed. PMID- 11476104 TI - Visual memories from nonvisual experiences. AB - Many common activities rely on spatial knowledge acquired from nonvisual modalities. We investigated the nature of this knowledge by having people look at a collection of objects on a desktop and manually reconstruct their arrangement, without vision, as though the display had been rotated by 0 degrees 45 degrees 90 degrees 135 degrees or 180 degrees relative to the view they could see. Performance on several measures of visual-spatial memory showed that participants had better visual memory for the view they had manually reconstructed than for the view they had studied visually for several minutes. These findings provide compelling new evidence that visual-spatial knowledge of very high fidelity can be acquired from nonvisual modalities, and reveal how, visual and nonvisual spatial information may even be confused in the brain. PMID- 11476105 TI - Knowing a word affects the fundamental perception of the sounds within it. AB - Understanding spoken language is an exceptional computational achievement of the human cognitive apparatus. Theories of how humans recognize spoken words fall into two categories: Some theories assume a fully bottom-up flow of information, in which successively more abstract representations are computed. Other theories, in contrast, assert that activation of a more abstract representation (e.g., a word) can affect the activation of smaller units (e.g., phonemes or syllables). The two experimental conditions reported here demonstrate the top-down influence of word representations on the activation of smaller perceptual units. The results show that perceptual processes are not strictly bottom-up: Computations at logically lower levels of processing are affected by computations at logically more abstract levels. These results constrain and inform theories of the architecture of human perceptual processing of speech. PMID- 11476106 TI - Desmosomes: structure and function in normal and diseased epidermis. AB - Desmosomes are important epidermal adhesion complexes that are characterized by a cell-specific expression of transmembrane cadherins and plaque-associated molecules. Desmosomes have so far, been implicated in three main disease types: autoimmune diseases that involve desmosome components (such as pemphigus vulgaris and pemphigus foliaceus), congenital diseases that affect intracellular calcium channels (such as Hailey-Hailey disease and Darier disease) and congenital diseases that directly affect desmosomal structural components. The identification of the first congenital defect affecting a desmosome component was in the gene for plakophilin I which caused an autosomal recessive skin fragility ectodermal dysplasia syndrome with skin, hair and nail defects. Subsequently, either a haploinsufficiency of desmoplakin or a defect in desmoglein 1 was found to underlie the autosomal dominant condition Striate Palmoplantar Keratoderma. In addition, plakoglobin has been shown to be defective in Naxos disease, which results in a cardiomyopathy and growth of abnormal hair. These findings pave the way for the discovery of further cell cohesion-related diseases and will help to greatly increase our understanding of the specific function of desmosome and other epithelial junction components. PMID- 11476107 TI - Stanozolol in chronic urticaria: a double blind, placebo controlled trial. AB - H1-type antihistamine drugs are mainstays in the management of chronic urticaria. For patients with refractory, chronic, idiopathic urticaria who have failed to benefit from conventional therapy, other safe therapeutic modalities are required. To evaluate the role of stanozolol as an adjunctive therapeutic agent with H1-antihistamine in refractory chronic idiopathic urticaria, we conducted this study. Fifty-eight patients with chronic refractory urticaria were enrolled in this trial and were randomly assigned to two groups (A and B). Patients in group A received 2 mg stanozolol twice daily along with cetrizine 10 mg daily. Patients in group B received cetrizine 10 mg daily and placebo tablets twice daily. The improvement was monitored by estimation of severity score. Of the 58 patients, 26 in group A and 24 in group B could be evaluated. At the end of 12 weeks, 17 patients in group A showed marked to complete resolution as compared to 7 patients in group B (chi-square p<0.01). The intention to treat analysis p value was a found to be <0.007. There was a highly significant decrease in mean severity score at 12 weeks (p<0.001) in group A patients. The present study demonstrated that stanozolol is an effective and safe adjuvant therapy for treatment of chronic refractory urticaria. PMID- 11476108 TI - Lipid analysis of peripheral blood monocytes in psoriatic patients using Fourier transform infrared microspectroscopy. AB - In psoriasis vulgaris, there are immunological abnormalities of T cells and monocytes. We previously demonstrated that monocytes in the peripheral blood of patients with psoriasis vulgaris are activated and produce an excess of inflammatory cytokines. It has long been suggested that fat metabolism is impaired in patients with this illness. In addition, it has been reported that macrophages activated by engulfing low density lipoprotein (LDL) immune complexes release large quantities of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta. Hence we hypothesized that the monocytes of psoriatic patients are activated by engulfing lipids and overproduce inflammatory cytokines. Therefore we measured both the serum and monocyte levels of lipids in the peripheral blood of psoriatic patients. At the same time, we calculated the psoriasis area and severity index (PASI) scores and analyzed their correlation with the lipid kinetics. The results showed that the serum cholesterol ester level and the cholesterol ester level in monocytes of psoriatic patients were significantly higher than those in healthy individuals. However, the cholesterol ester level in monocytes of patients with hyperlipidemia was also high, and there was no correlation between cholesterol ester level in monocytes of psoriatics and PASI scores. The cholesterol ester level in the monocytes of psoriatic patients was high, but this does not seem to play an important role in the pathogenesis of psoriasis. PMID- 11476109 TI - The significance of mast cells associated with basal cell carcinoma. AB - In the stroma of basal cell carcinoma (BCC), the numbers of mast cells are increased, but their significance remains unknown. In a retrospective histological study of 74 cases, we investigated the significance of mast cells related to BCC and the possible relationship between increased numbers of mast cells and clinical and histological parameters including the patients' age, sex, subtype of BCC, and severity of peritumoral inflammation. We found a significant increase in peritumoral mast cell numbers over those in uninvolved skin (p<0.0001). Morpheaform BCCs had significantly more mast cells than did the other variants (p=0.047). No significant relationship was found between mast cell number and the degree of peritumoral inflammation, patient age, or gender. Our results support previous reports indicating that mast cell numbers are increased in RCC, particularly in its morphoeic variant, implying a contributory role for mast cells in the aggressiveness of BCC. PMID- 11476110 TI - A case of childhood generalized pustular psoriasis treated with dapsone. AB - Acute generalized pustular psoriasis (GPP) is a rare, sometimes fatal, exudative form of psoriasis characterized by acute febrile pustular psoriasis. Unlike the adult forms, GPP in childhood tends to undergo a more benign course. It may resolve spontaneously with only supportive treatment, but may sometimes be life threatening and resistant to therapy. We report a case of GPP in 7-year-old girl treated successfully with dapsone. PMID- 11476111 TI - Lipodystrophia centrifugalis abdominalis infantilis: two cases from China. AB - We report the first two Chinese patients with lipodystrophia centrifugalis abdominalis infantilis in China. The first patient was a 3-year-old girl who developed an area of hyperpigmented skin depression on the right groin which spread centrifugally to the abdomen and right lower chest. Later, another lesion developed on the neck. The second patient was a 4-year-old girl who developed multiple areas of skin depression on her bilateral groin, periumbilical region, right axilla, and chest. The lesions also extended centrifugally. Erythematous margins and lymphadenopathy were present in both cases. Inflammatory changes were found in the subcutaneous fat. The clinical pictures and histologic features supported the diagnosis of LCAI in both cases. PMID- 11476112 TI - Hair follicle nevus occurring in frontonasal dysplasia: an electron microscopic observation. AB - We report a rare hair follicle nevus that occurred in a three-month-old Japanese boy with mild frontonasal dysplasia. It had been present since birth. Histologically, numerous tiny vellus hair follicles were found within the dermis. The constituent cells of these follicles showed the features of follicular germ cells under the electron microscope. The fibroblasts around the follicles were active and merged with the colloid substance. Many myofibroblasts were found in a collagenous stroma in the atrophic lesion of the frontonasal dysplasia. PMID- 11476113 TI - A case of folliculitis decalvans involving the beard, face and nape. AB - Folliculitis decalvans is a rare and chronic progressive hair disorder leading to scarring alopecia and atrophy. Its etiology is still unclear. It starts with areas of perifollicular erythema, and follicular papules and pustules spread peripherally, leaving central scarred patches of alopecia. We report a 27-year old man who presented with folliculitis decalvans involving the scalp, face, nape, and beard. There were pustules, crusts, and scarring alopecia on the scalp and beard. PMID- 11476114 TI - Localized dyschromatosis. PMID- 11476115 TI - A case of impetigo herpetiformis with unusual clinical features. PMID- 11476116 TI - Congenital eccrine angiomatous hamartoma: report of two patients. PMID- 11476117 TI - Post-kala-azar dermal leishmaniasis with an atypical presentation. PMID- 11476118 TI - Drug interactions with grapefruit juice: an update. PMID- 11476119 TI - A double-blind, randomized, prospective evaluation of the efficacy and safety of risperidone versus haloperidol in the treatment of schizoaffective disorder. AB - The relative efficacy and safety of risperidone versus haloperidol in the treatment of schizoaffective disorder was studied. Sixty-two patients (29 depressed type; 33 bipolar type) entered a three-site, randomized, double-blind, 6-week trial of risperidone (up to 10 mg/day) or haloperidol (up to 20 mg/day). Trained raters assessed baseline, weekly, and end-of-study levels of psychopathology with the Positive and Negative Syndrome Scale (PANSS), the 24 item Hamilton Rating Scale for Depression (HAM-D-24) and the Clinician Administered Rating Scale for Mania (CARS-M). The authors were unable to statistically distinguish between risperidone and haloperidol in the amelioration of psychotic and manic symptoms. In addition, there was no difference in worsening of mania between the two agents in either subgroup (i.e., depressed or bipolar subgroups). For the total PANSS, risperidone produced a mean decrease of 16 points from baseline compared with a 14-point decrease with haloperidol. For the total CARS-M scale, risperidone and haloperidol produced mean change scores of 5 and 8 points, respectively, and for the CARS-M Mania subscale, 3 and 7 points, respectively. Additionally, risperidone produced a mean decrease of 13 points from the baseline 24-item HAM-D, compared with an 8-point decrease with haloperidol. In those patients who had more severe depressive symptoms (i.e., HAM D baseline score >20), risperidone produced at least a 50% mean improvement in 12 (75%) of 16 patients in comparison to 8 (38%) of 21 patients receiving haloperidol. Haloperidol produced significantly more extrapyramidal side effects and resulted in more dropouts caused by any side effect. There was no difference between risperidone and haloperidol in reducing both psychotic and manic symptoms in this group of patients with schizoaffective disorder. Risperidone did not demonstrate a propensity to precipitate mania and was better tolerated than haloperidol. In those subjects with higher baseline HAM-D scores (i.e., >20), risperidone produced a greater improvement in depressive symptoms than haloperidol. PMID- 11476120 TI - Novel antipsychotics and severe hyperlipidemia. AB - Newer atypical antipsychotics demonstrate superior effectiveness, with a diminished incidence of extrapyramidal side effects compared with older typical antipsychotics, but they have been associated with the development of obesity and new-onset diabetes. A small number of reports documenting modest hypertriglyceridemia related to newer antipsychotics have implicated fluperlapine, clozapine, and, most recently, olanzapine. This study summarizes the results of 14 cases of severe hypertriglyceridemia (>600 mg/dL) associated with olanzapine and quetiapine therapy occurring among inpatients at Oregon State Hospital, including 7 patients whose serum triglyceride levels exceeded 1,000 mg/ dL. Four of these patients also developed new-onset diabetes. Nine cases occurred during the first 8 months of treatment, with three cases identified within 3 months of commencing olanzapine or quetiapine therapy. Weight gain in olanzapine and quetiapine groups was modest (12.3 lb and 8.5 lb, respectively) and did not correlate with the severity of hypertriglyceridemia. Biochemical causes for severe hypertriglyceridemia associated with novel antipsychotics are unclear, but clinical monitoring of serum lipids must be added to the concerns about the metabolic consequences of therapy with certain newer antipsychotic agents. PMID- 11476121 TI - Olanzapine as alternative therapy for patients with haloperidol-induced extrapyramidal symptoms: results of a multicenter, collaborative trial in Latin America. AB - Conventional antipsychotic agents can induce extrapyramidal symptoms (EPS) that may be alleviated by switching patients to novel agents such as olanzapine. Patients with schizophrenia and related disorders (ICD-10) who were taking haloperidol (N = 94; mean dose = 12.7 mg/day) and had EPS (Simpson-Angus Scale [SAS] > 3) were directly switched to 6 weeks of open-label olanzapine treatment (mean dose = 11.4 mg/day). There were significant mean improvements (p <0.001 for all measurements) from baseline to endpoint on the SAS (-9.69+/-5.33; percentage change, 87.2%), the Barnes Akathisia Scale (-1.00+/-1.19; percentage change, 82.5%), and the Abnormal Involuntary Movement Scale (-1.48+/-2.89; percentage change, 81.1%), and anticholinergic use decreased from 47.9% to 12.8% (mean baseline to endpoint change: -1.52+/-1.91-mg equivalents of benztropine; p < 0.001). Significant mean baseline to endpoint improvements (p < 0.001 for all measurements) were observed on the Positive and Negative Syndrome Scale (PANSS; 25.28+/-18.67; percentage change, 30.3%), the PANSS-extracted Brief Psychiatric Rating Scale (0-6 scale, -13.41+/-10.16; percentage change, 54.4%), and the Clinical Global Impressions Severity scale (-1.16+/-1.19; percentage change, 26.4%). Spontaneously reported treatment- emergent adverse events with a greater than 5% incidence were somnolence (16.0%), increased appetite (14.9%), weight gain (11.7%), headache (8.5%), anxiety (7.4%), dizziness (6.4%), and insomnia (5.3%). Criteria for a successful switch were met by 90.5% of patients. Psychotic symptom exacerbation was experienced by 30.9% of patients at any time during the study and by 11.7% of patients at endpoint. Results suggest that a direct switch to olanzapine is a therapeutic option when patients with haloperidol-induced EPS are unable to tolerate a more gradual switch. PMID- 11476122 TI - Myocarditis related to clozapine treatment. AB - Myocarditis has in several case reports been associated with use of clozapine. Eight cases of myocarditis during treatment with clozapine that were submitted to the Swedish Adverse Drug Reaction Advisory Committee and 18 cases that were reported in the literature are summarized. As part of the routine signal detection process on the World Health Organization (WHO) Program on International Drug Monitoring database, which contains more than two million case reports of spontaneously reported suspected adverse drug reactions, a Bayesian confidence propagation neural network (BCPNN) is used. This article also shows the retrospective output of the BCPNN over time for clozapine and myocarditis and discusses its implications. In 19 (79%; duration of treatment not stated for 2 patients) of 24 patients with myocarditis, the symptoms occurred within the first 6 weeks of clozapine treatment. Many patients shared a similar clinical course, with symptoms such as an influenza-like illness, fever, sinus tachycardia, hypotension, chest discomfort, and heart failure. The reaction was fatal in 12 (46%) of these patients. The other patients generally had a prompt recovery. By using the BCPNN technique, a quantitative association between clozapine and myocarditis was demonstrated, and the association might have been high-lighted for clinical review in 1994 had this BCPNN method been in use at the WHO center at the time. Myocarditis seems to be a rare and potentially lethal adverse effect of clozapine. Admittance for observation, interruption of the clozapine treatment, and treatment with corticosteroids should be considered for patients in whom this reaction is suspected. PMID- 11476123 TI - A double-blind, randomized comparison of the efficacy and safety of intramuscular injections of olanzapine, lorazepam, or placebo in treating acutely agitated patients diagnosed with bipolar mania. AB - There are no rapid-acting intramuscular formulations of atypical antipsychotics available for quickly calming an agitated patient with bipolar disorder. In this study, 201 agitated patients with bipolar mania were randomly assigned to receive one to three injections of the atypical antipsychotic olanzapine (10 mg, first two injections; 5 mg, third injection), the benzodiazepine lorazepam (2 mg, first two injections; 1 mg, third injection), or placebo (placebo, first two injections; olanzapine, 10 mg, third injection) within a 24-hour period. Agitation was measured at baseline, every 30 minutes for the first 2 hours, and at 24 hours after the first injection using the Positive and Negative Syndrome Scale-Excited Component subscale and two additional agitation scales. At 2 hours after the first injection, patients treated with olanzapine showed a significantly greater reduction in scores on all agitation scales compared with patients treated with either placebo or lorazepam. At 24 hours after the first injection, olanzapine remained statistically superior to placebo in reducing agitation in patients with acute mania, whereas patients treated with lorazepam were not significantly different from those treated with placebo or olanzapine. Furthermore, no significant differences among the three treatment groups were observed in safety measures, including treatment-emergent extrapyramidal symptoms, the incidence of acute dystonia, or QTc interval changes. These findings suggest that intramuscular olanzapine is a safe and effective treatment for reducing acute agitation in patients with bipolar mania. PMID- 11476124 TI - CYP1A2 activity as measured by a caffeine test predicts clozapine and active metabolite steady-state concentrationin patients with schizophrenia. AB - Clozapine is an atypical antipsychotic drug and displays efficacy in 30% to 60% of patients with schizophrenia who do not respond to traditional antipsychotics. A clozapine concentration greater than 1,150 nmol/L increases the probability of antipsychotic efficacy. However, plasma clozapine concentration can vary more than 45-fold during long-term treatment. The aim of this study was to assess the contribution of CYP1A2 to variability in steady-state concentration of clozapine and its active metabolite norclozapine. Patients with schizophrenia or schizoaffective disorder were prospectively monitored during clozapine treatment (N = 18). The in vivo CYP1A2 activity was measured using the caffeine metabolic ratio (CMR) in overnight urine. Trough plasma samples were drawn after at least 5 days of treatment with a constant regimen of clozapine. A significant negative association was found between the CMR and the dose-corrected clozapine (r(s) = 0.87,p < 0.01) and norclozapine (r(s) = -0.76,p < 0.01) concentrations. Nonsmokers displayed a higher clozapine (3.2-fold) and norclozapine (2.3-fold) concentration than smokers (p < 0.05). Furthermore, there was marked person-to person variation in CYP1A2 activity during multiple-dose clozapine treatment (coefficient of variation = 60%). Age, weight, serum creatinine, and grapefruit juice consumption did not significantly contribute to variability in clozapine and norclozapine concentration (p > 0.05). In conclusion, CYP1A2 is one of the important contributors to disposition of clozapine during multiple-dose treatment. Although further in vitro experiments are necessary, the precise metabolic pathways catalyzed by CYP1A2 seem to be subsequent to the formation of norclozapine, hitherto less recognized quantitatively important alternate disposition routes, or both. From a clinical perspective, an environmentally induced or constitutively high CYP1A2 expression can lead to a decrease in steady state concentration of clozapine as well as its active metabolite norclozapine. Thus, interindividual variability in CYP1A2 activity may potentially explain treatment resistance to clozapine in some patients. CYP1A2 phenotyping with a simple caffeine test may contribute to individualization of clozapine dosage and differentiate between treat ment noncompliance and high CYP1A2 activity. PMID- 11476125 TI - An evaluation of risperidone drug interactions. AB - Risperidone, an atypical antipsychotic drug, is widely used in the treatment of psychoses associated with schizophrenia, Alzheimer's disease, and other psychiatric disorders. Polypharmacology is a necessary condition for the optimal treatment of many patients with comorbid psychiatric and medical illness. One concern raised by the widespread use of multiple concurrent pharmacotherapies is the potential for drug-drug interactions to adversely affect patient outcome. Accordingly, the biomedical literature was reviewed for reports of drug interactions involving risperidone, and the clinical significance of each report was evaluated. Additionally, the potential for risperidone to participate in drug interactions was evaluated by considering the drug's pharmacokinetic properties. Controlled studies and case reports indicate that risperidone has a low potential for metabolic drug interactions. Drugs that inhibit cytochrome P450 (CYP) 2D6 or induce or inhibit CYP3A4 may alter risperidone plasma concentrations, but the clinical significance of such interactions seems to be minimal. Adherence to a few guidelines for the design of dosage regimens should limit the effect of drug drug interactions on patient status and contribute to optimal pharmacotherapy with risperidone. PMID- 11476126 TI - Fluoxetine in the prevention of depressive recurrences: a double-blind study. AB - Optimal outcomes from depression treatment are long-term recovery and, in the case of recurrent depression, prevention of new episodes. However, few data are available concerning the long-term efficacy of antidepressants in prophylactic treatment to prevent recurrences of depression. The efficacy and safety of fluoxetine 20 mg/day was evaluated in reducing the number of depressive episodes and in extending the time free of symptoms in patients with recurrent unipolar major depression. Patients with recurrent unipolar major depression according to DSM-III-R criteria and who responded to 32 weeks of open-label fluoxetine were randomly assigned to receive fluoxetine 20 mg/day (N = 70) or placebo (N = 70) for 48 weeks of double-blind maintenance treatment. Outcome measures were the percentage of recurrences and time to recurrence. Safety assessments included treatment-emergent adverse events, reasons for discontinuation, vital signs, and laboratory measures. Fluoxetine was associated with a statistically significantly smaller percentage of patients who had a recurrence compared with placebo (20% vs. 40%; chi2 analysis, p = 0.010). The symptom-free period was significantly longer for patients treated with fluoxetine versus placebo (295 vs. 192 days; Kaplan-Meier estimates, log-rank test, p = 0.002). Treatments were well tolerated during maintenance treatment. The only statistically significant difference in adverse events between treatment groups was anxiety, which was more frequent in the placebo group (fluoxetine, 12.9% vs. placebo, 30%; chi2 analysis, p = 0.013). Two placebo-treated patients and no fluoxetine-treated patients were withdrawn because of adverse events. In conclusion, fluoxetine at 20 mg/day was effective and well tolerated for the prophylactic treatment of recurrent unipolar major depression. PMID- 11476127 TI - Mirtazapine versus venlafaxine in hospitalized severely depressed patients with melancholic features. AB - The aim of this multicenter, randomized, double-blind, 8-week study was to compare the antidepressant efficacy and tolerability of mirtazapine and venlafaxine in the treatment of hospitalized patients with DSM-IV diagnosis of severe depressive episode with melancholic features. Patients with a baseline score of > or = 25 on the 17-item Hamilton Rating Scale for Depression (HAM-D-17) were randomly assigned to receive treatment with either mirtazapine (N = 78, 15 60 mg/day) or venlafaxine (N = 79, 75-375 mg/day, twice a day) in a rapid up titration schedule. Efficacy was assessed with the Montgomery-Asberg Depression Rating Scale (MADRS), HAM-D-17, and Clinical Global Impression scale, and quality of life was assessed with the Quality of Life, Enjoyment, and Satisfaction Questionnaire and Quality of Life in Depression Scale. Tolerability was assessed with the Utvalg for Kliniske Undersogelser (UKU) side effect scale and by reporting adverse events. Both drugs were effective in reducing overall symptoms of depression, showing substantial reductions in group mean MADRS scores (-20.1 for mirtazapine and -17.5 for venlafaxine) and HAM-D-17 scores (-17.1 for mirtazapine and -14.6 for venlafaxine) at the end of the treatment. Although not statistically significant, at all assessment times higher percentages of patients treated with mirtazapine were classified as responders (> or =50% reduction) on the HAM-D (at endpoint, 62% vs. 52%) and MADRS (at endpoint: 64% vs. 58%). Likewise were the percentages of remitters (HAM-D score < or =7; MADRS score < or =12) also higher in the mirtazapine group. A statistically significant difference favoring mirtazapine was found on the HAM-D Sleep Disturbance factor at all assessment points (p < or = 0.03). Both treatments were well tolerated. Although slightly more subjects treated with mirtazapine reported at least one adverse event, a statistically significantly higher percentage of patients treated with venlafaxine (15.3%) than mirtazapine (5.1%) dropped out because of adverse events (p = 0.037). Quality of life improved in both treatment groups. In this study, treatment with mirtazapine resulted in a trend toward more responders and remitters than treatment with venlafaxine and in significantly fewer dropouts as a result of adverse events. PMID- 11476128 TI - Doxepin plasma concentrations: is there really a therapeutic range? AB - Despite the introduction of numerous new agents, tricyclic antidepressants remain an important option for the treatment of depression. Doxepin is still in wide use, and determining its concentration is a standard procedure in many psychiatric clinics. Some widely cited reviews indicate a therapeutic range from 150 to 250 ng/mL (parent plus desmethyl metabolite). The vast majority of the authors' patients fell short of these concentrations under customary doses. To resolve this issue, the authors' serum-level databank was analyzed, a questionnaire was sent to U.S. and German psychiatric university departments and laboratories, and the literature was reviewed. The main results were the following: (1) Only 9% of all samples analyzed (N = 217) displayed plasma levels (high-performance liquid chromatography) between 150 and 250 ng/mL; 88% were subtherapeutic. The mean doxepin + desmethyldoxepin steady-state serum concentration was 89+/-75 ng/mL (N = 32, doxepin >3 weeks). The mean daily dose was 143+/-30 mg. There was no correlation between concentrations and improvement. (2) A wide variety of recommendations is given by the different university departments (10-1,000 ng/mL). (3) According to the studies published to date, there is not enough evidence for recommending a therapeutic range. The preliminary suggestions given in some influential reviews have been widely adopted without critical re-evaluation. Compared with the concentrations found in the original studies, the therapeutic ranges suggested are too high. A methodologically sound study to determine a therapeutic range is required for the rational monitoring of this drug. Meanwhile, a preliminary working range of 50 to 250 ng/mL is proposed on the basis of critical reassessment of published data. PMID- 11476129 TI - Clomipramine versus haloperidol in the treatment of autistic disorder: a double blind, placebo-controlled, crossover study. AB - Clomipramine, haloperidol, and placebo were compared with baseline in the treatment of autism, and overall outcome, specific symptoms, and side effects were examined. It was hypothesized that clomipramine would be better tolerated than haloperidol and prove superior on a measure of stereotypy. Individuals with a DSM-IV diagnosis of autistic disorder (mean age, 16.3 years; range, 10-36 years) were randomly assigned, by using a Latin square design, to the following 7 week trials: placebo, clomipramine (mean daily dose, 128.4 mg; range, 100-150 mg), or haloperidol (mean daily dose, 1.3 mg; range, 1-1.5 mg). Data on 36 subjects were analyzed and taken together; the results favored haloperidol. In those patients who were able to complete a full therapeutic trial, clomipramine proved comparable to haloperidol in terms of improvement compared with baseline. However, significantly fewer individuals receiving clomipramine versus haloperidol were able to complete the trial (37.5% vs. 69.7%, respectively) for reasons related to both side effects and efficacy or behavior problems. In the intent-to-treat sample, which is perhaps more clinically relevant, only haloperidol proved superior to baseline on a global measure of autistic symptom severity, as well as specific measures for irritability and hyperactivity. Clomipramine did not seem more effective on a measure of stereotypy, nor was it better tolerated. PMID- 11476130 TI - Haloperidol blood levels in acute mania with psychosis. AB - In this study, the authors examined the relationship between steady-state haloperidol blood levels and clinical response in patients with acute psychotic mania. Fifty-four inpatients with acute mania were randomly assigned to receive either haloperidol 25 mg/day or haloperidol 5 mg/day. Each subject also received a concomitant medication: lorazepam 4 mg/day, lithium, or placebo. The relationship between steady-state haloperidol blood levels and clinical improvement was studied using analysis of covariance. There was wide interindividual variation in the haloperidol blood level-dose ratio. Haloperidol blood levels (log-transformed) were found to significantly correlate with clinical response in acute mania. Low-dose haloperidol with concomitant lithium may produce an optimal response in acute mania. Haloperidol blood levels may be clinically useful in identifying patients who are nonresponsive because of low drug levels and, hence, in enhancing optimal haloperidol dosing for acute mania with psychosis. PMID- 11476131 TI - Olanzapine for schizophrenia refractory to typical and atypical antipsychotics: an open-label, prospective trial. AB - The role of olanzapine in treatment-resistant schizophrenia is still unresolved. This article presents an open-label, prospective, 14-week trial with olanzapine in patients with schizophrenia and schizoaffective disorder selected for unambiguous resistance to either clozapine or risperidone and to typical antipsychotics. Forty-three inpatients (mean age, 41.6 years; mean duration of illness, 21.7 years) were enrolled and treated after cross-titration from their previous antipsychotic treatment with olanzapine 10 to 40 mg daily without any concomitant antipsychotic medication. Patients were evaluated with the Positive and Negative Syndrome Scale (PANSS), the Clinical Global Impressions Scale, and the Extrapyramidal Symptom Rating Scale. The change with olanzapine treatment was associated with a PANSS total score improvement of 3.7 (SD = 15.6; not significant). There was a significant improvement for the PANSS cognitive and depression/anxiety factors, whereas the PANSS excitement factor worsened. The improvement rate was superior in patients receiving olanzapine doses higher than 20 mg. A total of 16.7% of patients reached response criteria set forth by a previous study. There was a significant decrease in extrapyramidal side effects (t = 2.04; p < 0.05) and statistically significant, yet modest, weight gain. These results indicate that olanzapine is only modestly effective in these severely treatment-resistant patients with schizophrenia. However, a trial with olanzapine can be recommended in these patients before moving to augmentation strategies, given the lack of proven alternatives and the observation that 16.7% of patients reached the response criteria. PMID- 11476132 TI - An open study of buspirone augmentation of neuroleptics in patients with schizophrenia. PMID- 11476133 TI - Comments on "Treating Charles Bonnet syndrome: understanding inconsistency". PMID- 11476134 TI - Maternal exposure to lorazepam and anal atresia in newborns: results from a hypothesis-generating study of benzodiazepines and malformations. PMID- 11476135 TI - Arthritis and serotonergic antidepressants. PMID- 11476136 TI - Panic attacks associated with the use of topiramate. PMID- 11476137 TI - Topiramate in venlafaxine-induced visual hallucinations in an obese patient with a posterior cerebral artery infarction. PMID- 11476138 TI - First break of mania associated with topiramate treatment. PMID- 11476139 TI - Winning hearts and minds. PMID- 11476140 TI - Readmission of patients to hospital: still ill defined and poorly understood. PMID- 11476141 TI - Readmission of elderly patients to hospital: still ill-defined and poorly understood--a response. PMID- 11476142 TI - Readmissions--are we asking too much? PMID- 11476143 TI - Validating risk-adjusted surgical outcomes: chart review of process of care. AB - OBJECTIVE: The primary purpose of this study was to validate risk-adjusted surgical outcomes as indicators of the quality of surgical care at US Department of Veterans Affairs (VA) hospitals. The secondary purpose was to validate the risk-adjustment models for screening cases for quality review. DESIGN: We compared quality of care, determined by structured implicit chart review, for patients from hospitals with higher and lower than expected operative mortality and morbidity (hospital-level tests) and between patients with high and low predicted risk of mortality and morbidity who died or developed complications (patient-level tests). SUBJECTS: 739 general, peripheral vascular and orthopedic surgery cases sampled from the 44 VA hospitals participating in the National VA Surgical Risk Study. MAIN OUTCOME MEASURES: A global rating of quality of care based on chart review. RESULTS: Ratings of overall quality of care did not differ significantly between patients from hospitals with higher and lower than expected mortality and morbidity. On some of the secondary measures, patient care was rated higher for hospitals with lower than expected operative mortality. At the patient level of analysis, those who died or developed complications and had a high predicted risk of mortality or morbidity were rated higher on quality of care than those with a low predicted risk of adverse outcome. CONCLUSIONS: The absence of a relationship between most of our measures of process of care and risk-adjusted outcomes may be due to an insensitivity of chart reviews to hospital-level differences in quality of care. Site visits to National VA Surgical Risk Study hospitals with high and low risk-adjusted mortality and morbidity have detected differences on a number of dimensions of quality. The patient-level findings suggest that the risk-adjustment models are useful for screening adverse outcome cases for quality of care review. PMID- 11476144 TI - Issues in the assessment of continuous quality improvement implementation in health care organizations. AB - OBJECTIVE: This paper has two primary aims. First, it examines the need for improved assessment of continuous quality improvement implementation. Second, it analyzes current worldwide measures and studies of continuous quality improvement implementation. METHOD: A comprehensive literature review was conducted which included all published (English language) studies of organization-wide continuous quality improvement implementation. RESULTS: Analysis of the content and research methods incorporated into current measures of continuous quality improvement implementation used worldwide supports a strong consensus regarding the major criteria that need to be addressed. However, there are still promising areas for future research, namely increased use of criteria other than the Baldrige categories, increased focus upon financial variables, improved measures of implementation stage/phase and the use of different types of respondents from multiple organizational levels. CONCLUSION: Increased understanding of the empirical benefits and costs of continuous quality improvement in health care organizations is heavily contingent upon the continued development and improvement of measures of continuous quality improvement implementation. PMID- 11476145 TI - Quality assessment of oral anticoagulant treatment in the Beer-Sheba district. AB - OBJECTIVE: To evaluate the adequacy of oral anticoagulation therapy in patients with either prosthetic heart valves or atrial fibrillation. DESIGN: Adequacy of anticoagulation therapy of patients treated with warfarin was determined before and after implementation of an improvement programme. SETTING: The central haematology laboratory of the Soroka Medical Center, Beer-Sheba Israel. STUDY PARTICIPANTS: One hundred and ten patients treated with chronic anticoagulation therapy were evaluated by measuring the international normalized ratio of the prothrombin time before and after implementation of an improvement plan aimed at improving anticoagulation control. INTERVENTION: A programme that included physician and patient education and procedural changes covering all aspects of anticoagulation therapy was implemented. MAIN OUTCOME MEASURE: The percent of international normalized ratio tests below, within and above the therapeutic range was determined. RESULTS: Prior to implementation of an improvement plan, only 32% of the measured international normalized ratio values were within the therapeutic range. 16% were above and 52% were below therapeutic range. Improvement intervention led to an increase in the proportion of international normalized ratio tests that fell within the therapeutic range from 32% to 43.2%. CONCLUSIONS: Improvement in the control of anticoagulation therapy of patients receiving long-term warfarin treatment is possible. A combined effort involving the community, physician and patient is required. PMID- 11476146 TI - Evaluating satisfaction with maternity care in women from minority ethnic communities: development and validation of a Sylheti questionnaire. AB - OBJECTIVE: To develop a reliable and valid questionnaire to evaluate satisfaction with maternity care in Sylheti-speaking Bangladeshi women. DESIGN: Two-stage, psychometric study. Firstly, focus groups, in-depth interviews and iterative methods for translation and cultural adaptation were used to develop a Sylheti questionnaire, called the survey of Bangladeshi women's experience of maternity services from an English language questionnaire. Secondly, quantitative psychometric methods were used to field test and evaluate the acceptability, reliability and validity of this questionnaire. SETTING: Four hospitals providing maternity services in London, UK. STUDY PARTICIPANTS: Two hundred and forty-two women from the London Bangladeshi communities, who were in the antenatal (at least 4 months pregnant) or postnatal phase (up to 6 months after delivery). Women spoke Sylheti; a language with no accepted written form. Two purposive samples of 40 women in the antenatal or postnatal phase, one convenience sample of six women in the antenatal phase and three consecutve samples of 60 women in the postnatal phase participated in stage one. In stage two, 135 women (main sample) completed the questionnaire two months after delivery (82% response rate); 50 women (retest sample) from the main sample completed a second questionnaire two weeks later (96% response rate). MAIN OUTCOME MEASURES: Women's views about maternity care elicited by qualitative methods and measured quantitatively using the survey of Bangladeshi women's experience of maternity services. RESULTS: The 121-item questionnaire was acceptable to women and showed good internal consistency (Cronbach's alphas 0.76-0.91), stability (test-retest reliability 0.72-0.84) and construct validity (e.g. able to detect group differences). CONCLUSION: By combining qualitative and quantitative methods, it is possible to adapt an instrument to provide an acceptable, reliable and valid Sylheti questionnaire. The approach taken in developing this questonnaire provides a model for developing outcome measures for use with other minority ethnic communities. PMID- 11476147 TI - Resident physician management of Hispanic and non-Hispanic white patients on antidepressants. AB - OBJECTIVE: The purpose of this study was to examine how well resident physicians monitored Hispanic and non-Hispanic white patients who were prescribed antidepressant medication. METHOD: Retrospective and prospective review of patients' medical records. SETTING: Family practice and internal medicine clinics at the University of New Mexico Health Sciences Center. PARTICIPANTS: Twenty-six resident physicians and 109 of their Hispanic and non-Hispanic white patients who were prescribed antidepressant medication when recruited into the study between March and December 1995. MAIN OUTCOME MEASURES: (i) Whether a physician recorded an appropriate diagnosis in the patient's chart, scheduled a follow-up visit and saw the patient for a follow-up visit within four weeks of the antidepressant being prescribed, and (ii) whether a physician recorded an adequate treatment plan, a discussion of side-effects and a discussion of how well the medication was working on the date the patient was enrolled in the study. RESULTS: Twenty seven percent of patients were prescribed antidepressant medication. Less than half of the patients who were prescribed an antidepressant had a follow-up visit scheduled and only about one-third of patients were seen by their physician within one month of the antidepressant being prescribed. Physicians documented an adequate treatment plan in the charts of 51.1% of patients, a discussion of side effects in 11.1% of charts and an assessment of how well the medication was working in 33.3% of charts. Younger patients and patients in better emotional health were more likely to have an adequate treatment plan documented in their chart. Patients in poorer physical health were more likely to have an adequate treatment plan documented in their chart along with a description of the effectiveness of the medication. Hispanic and non-Hispanic white patients who were prescribed antidepressants were monitored equally well by their primary care physicians. CONCLUSION: Primary care resident physicians need further training on the importance of monitoring patients on antidepressant medication. PMID- 11476148 TI - Perceived effectiveness of diagnostic and therapeutic guidelines in primary care quality circles. AB - OBJECTIVE: The main objectives of this study were to implement quality circle programs among general practitioners and to evaluate this quality management tool as a way to develop clinical guidelines in general practice. DESIGN: The quality circle program was evaluated within a formative and summative evaluation design by both participants and moderators for a period of 18 months using structured questionnaires. At time one, participants were asked about their goals and current job satisfaction, and rated the perceived effectiveness and the usefulness of predefined guidelines of each quality circle meeting. At time two, participants and moderators reported again about their achieved goals and job satisfaction. SETTING AND STUDY PARTICIPANTS: Two hundred and forty-three general practitioners in a district of South Germany (Sudbaden), in 25 quality circle groups participated. MAIN MEASURES: Demographic variables of the participating physicians, quality circle goals, job satisfaction, usefulness of guidelines and perceived effectiveness of the quality circle process were collected. RESULTS: One hundred and six quality circle meetings were evaluated. When asked to rank the goals of quality circle work, participants provided the highest rankings for improvement of the doctor-doctor relationship, agreeing on consensus for diagnostic procedures and therapy management, and developing local guidelines. The comparison between time one and time two ratings provided evidence for an increase in overall job satisfaction. Higher benefit is correlated with more regular participation in quality circle meetings. CONCLUSION: Working with predefined guidelines is both feasible and effective in quality circles and may provide a starting point for developing guidelines in primary care. There is some empirical evidence that participating in quality circles may increase general practitioners' job satisfaction. Further studies using intervention and control group designs should investigate whether quality circles really improve daily practice through clinical audit and benchmarking techniques. PMID- 11476149 TI - Clinical service standards at academic health centers. AB - OBJECTIVE: To describe the content and variability for clinical service standards related to quality of care among a convenience sample of academic health centers. DESIGN: We used the membership of the University HealthSystem Consortium, an alliance of academic health centers in the United States for clinical services, to survey electronically 53 of these centers regarding clinical service standards. The survey evaluated service standards in four areas; general communications, communications between physicians, ambulatory and inpatient clinical services and administrative standards. RESULTS: Thirty-four institutions responded to the survey (64%). Of these, 16 (47%) had clinical service standards, while the remaining 18 (53%) had not established formal standards. A few of the centers had established standards for patient communications, such as policies for answering telephones by staff. More had developed standards for communications between physicians and most centers had established standards for appointment availability, especially for urgent visits. However, clinical service standards were less typical for inpatient consultative or diagnostic services. A small number of the academic health centers had standards for hours of operation and for handling administrative matters, such as patient complaints. For many clinical service standards at the centers, there were notable variations (e.g. non-urgent primary care visits ranged 3-14 days). CONCLUSION: Some academic health centers have developed and implemented patient-centered clinical service standards for diverse areas of practice, however, the standards used appear to vary for some aspects of care, but not for others. PMID- 11476150 TI - An evaluation of the QSP and the QPP: two methods for measuring patient satisfaction. AB - BACKGROUND: Patient satisfaction is a function of several variables addressing reasons why it is important to use methods in which these different factors can be isolated and their importance analysed. OBJECTIVE: In this project, two methods using this approach were used: the 'Quality from the Patient's Perspective' and the 'Quality, Satisfaction, Performance' models. The aim of the present study is to evaluate these two different methods with respect to application, strengths and weaknesses. DESIGN: In the Quality from the Patient's Perspective model, the patient judges the different domains in two dimensions: perceived reality and subjective importance. The Quality, Satisfaction, Performance model uses a multivariate analysis to capture the patient's priorities. Four hundred and sixty forms for each model were distributed to a random sample of patients at the Department of Obstetrics and Gynecology at Karolinska Hospital. MAIN MEASURES: The quality factors 'treatment by the nurse', 'participation', 'information', 'environment' and 'accessibility' were measured. RESULTS: On both forms, 'medical care', 'treatment by the doctor' and 'access to nursing treatment' received high scores in perceived reality' while 'accessibility' and 'participation' received low scores. 'Subjective importance' measured directly and indirectly, respectively, in the two models showed high values for 'medical care' and 'treatment by the doctor'. CONCLUSION: The advantages of the Quality from the Patient's Perspective model are that it has a comprehensive and solid question bank. The Quality, Satisfaction, Performance model's advantage is its immediate usefulness and its clear graphic presentation. An integration and further development of these two approaches may prove useful. PMID- 11476151 TI - Communication breakdowns combine to cause system failure. PMID- 11476152 TI - Predictors of satisfaction with surgical treatment. PMID- 11476153 TI - Metabolic capacity and interindividual variation in toxicokinetics of styrene in volunteers. AB - The aim of the present study was to assess the interindividual variation in styrene toxicokinetics and to correlate this variation with the individual metabolic capacity for cytochrome P450 (CYP), CYP2E1, CYP1A2 and CYP2D6. Twenty male volunteers were exposed on separate occasions to 104+/-3 and 360+/-20 mg/m3 of styrene for 1 h while performing 50 W physical exercise on a bicycle ergometer. Styrene concentrations in blood and mandelic (MA) and phenylglyoxylic acid (PGA) in urine were measured. The metabolic capacity was assessed by phenotyping with chlorzoxazone (CYP2E1), caffeine (CYP1A2), dextromethorphan (CYP2D6) and antipyrine (CYP450). In addition, for the main styrene-metabolising enzyme, CYP2E1, genotyping for the genetic polymorphisms of the gene was performed. The average pulmonary retention of styrene was 62 +/- 7% at both exposure concentrations, and the 24-h excretion of MA and PGA accounted for 58% of the dose at both concentrations. The interindividual variation in styrene kinetics ranged from 19% for the terminal half-life (t(1/2,beta)) of styrene to 41% for the cumulative excretion of MA and PGA. However, no correlation between the apparent blood clearance of styrene (CLapp), t(1/2,beta) of styrene or excretion of MA and PGA on one hand, and the individual metabolic capacity on the other hand was found. Although other explanations cannot be excluded, this lack of correlation might be due to the high apparent blood clearance (1.4 l/min) of styrene, indicating that styrene metabolism is liver-blood-flow-dependent. PMID- 11476154 TI - Activation of human neutrophils by chlordane: induction of superoxide production and phagocytosis but not chemotaxis or apoptosis. AB - We have recently reported that human neutrophils are important targets of different xenobiotics, including chemicals of environmental concern. In the present study, we found that chlordane was not toxic for human neutrophils incubated for up to 24 h with concentrations ranging from 0.1 to 50 microg/ml. Chlordane was found to induce neutrophil superoxide production (O2-) in a concentration-dependent fashion and its potency to induce this response was found to be similar to phorbol 12-myristate 13-acetate (PMA), a classical neutrophil agonist. The use of different transduction signal inhibitors (genistein, pertussis toxin, staurosporine, and calphostin C) indicates that, as for PMA, chlordane induces O2- production via protein kinase C (PKC). In this respect, staurosporine and calphostin C were found to inhibit chlordane- and PMA-induced O2- production by 65% and 72%, and by 83% and 85%, respectively. Chlordane was also found to significantly enhance neutrophil phagocytosis of opsonized sheep red blood cells (SRBCs). Despite these effects, chlordane did not alter neutrophil apoptosis as assessed by cytology (Diff-Quick staining) and by flow cytometry (CD-16 expression). In addition, chlordane did not alter neutrophil chemotaxis (48-well Boyden chamber). Cells were, however, responsive as they were activated by the well-characterized interleukin (IL)-8 chemokine. We conclude that chlordane can activate O2- production by a PKC-dependent mechanism and induce phagocytosis without altering chemotaxis and apoptosis. Chlordane must be added to a growing list of environmental contaminants that share some pro inflammatory properties. PMID- 11476155 TI - Reduction of uranium transfer by local chelation in simulated wounds in rats. AB - The aim of the paper is to develop a new approach to treat uranium-contaminated wounds. The efficacy of a local uranium chelator, carballylic amido bis phosphonic acid (CAPBP) was assessed using two different uranium compounds. Rats were contaminated by intramuscular injections of uranyl nitrate or an industrial U04 compound to simulate wound contamination. CAPBP was injected intramuscularly (i.m.) or intraperitoneally (i.p.) at a dosage of 30 micromol kg(-1). In one experiment, the local administration of CAPBP was combined with a systemic administration of ethane-1-hydroxy-1,1-biphosphonate (EHBP). The local CAPBP treatment resulted in increased retention of uranium at the wound site: about 30% for uranyl nitrate or U04 after the first day and about 15% of UO4 after the third day. Consequently, it reduced uranium translocation into the blood and deposition in the kidneys and bone. The combined treatment reduced the uranium deposits in the kidneys, bone and carcass to about one-half of those observed in controls 3 days after U04 contamination. The local CAPBP treatment increased the interval of time between contamination and uranium deposit in the target organs. Thus, it can increase the efficacy of nonspecific local treatments or specific systemic treatments. It could be given rapidly through spray or gel after an accident. PMID- 11476156 TI - Effects of Vepacide (Azadirachta indica) on aspartate and alanine aminotransferase profiles in a subchronic study with rats. AB - The aim of this study was to ascertain the long-term effects of Vepacide, a neem based pesticide on biochemical profiles. Albino Wistar rats were treated orally with 80 (low), 160 (medium) and 320 mg/kg (high) doses of Vepacide in coconut oil for 90 days. Control rats received the same volume of the vehicle. Vepacide caused increase of aspartate and alanine aminotransferase in serum, kidney and lung, and these enzymes decreased in liver in both male and female rats when measured after 45 and 90 days of treatment. The two-way analysis of variance (ANOVA) showed that the alterations in these enzymes were dose- and time dependent. Sexual dimorphism was observed when male rats were compared with female rats (Student t-test at P< 0.05). Positive correlation was observed with regard to these enzymes between serum, kidney and lung, whereas in the case of serum and liver, a negative correlation was recorded. These enzyme profiles elucidate that they increased in serum with simultaneous decrease in liver, indicating necrosis of liver, whereas in other tissues, the level of enzymes increased, showing an adaptive mechanism due to the chemical stress. The affected enzymes were recovered to normal conditions after 28 days of post-treatment (withdrawal study). Due to the Vepacide treatment, lung was more affected followed by liver and kidney. This study has indicated that these enzymes could be useful as biomarkers for the insult of any toxicant. Besides, they can also help in predictive toxicology. PMID- 11476157 TI - Taurine has a protective effect against thioacetamide-induced liver cirrhosis by decreasing oxidative stress. AB - Thioacetamide (TAA) administration (0.3 g/l of tap water for a period of 3 months) to rats resulted in hepatic cirrhosis as assessed by biochemical and histopathological findings. This treatment caused an increase in the levels of malondialdehyde (MDA) and diene conjugates (DCs) and a decrease in the levels of glutathione (GSH), vitamin E, vitamin C and the activities of glutathione peroxidase (GSH-Px) in the liver of rats. Superoxide dismutase (SOD) activities were unchanged. Taurine (2% w/w, added to the chow diet) was administered together with TAA (0.3 g/l of drinking water) for 3 months. Taurine was found to decrease TAA-induced hepatic lipid peroxidation and to increase TAA-depleted vitamin E levels and GSH-Px activities. Histopathological findings also suggested that taurine has an inhibitive effect on TAA-induced hepatic cirrhosis. These results indicate that taurine treatment has a protective effect against TAA induced liver cirrhosis by decreasing oxidative stress. PMID- 11476158 TI - Alcohol and aldehyde dehydrogenase activity in the stomach and small intestine of rats poisoned with methanol. AB - The activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were measured with fluorogenic naphthaldehydes in the stomach and small intestine homogenates of rats dosed with 6 g methanol/kg bw after 6, 12, 24 h and 2, 5, 7 days. After intoxication with a sublethal dose, the ADH activity measured with these naphthaldehydes and ALDH activities in the stomach and small intestine were significantly decreased. This inhibition is stronger in the stomach and probably depends on cell damage and protein denaturation. We conclude that the activity measured with 6-methoxy-2-naphthaldehyde (MONAL-62) may be due to the activity of rat ADH-1 isoenzyme, and the activity detected with 4-methoxy-1-naphthaldehyde (MONAL-41) to the activity of rat ADH-2 isoenzyme. PMID- 11476159 TI - Ambient particulate matter induces relaxation of rat aortic rings in vitro. AB - Epidemiological studies have shown an association between ambient levels of particulate matter (PM) and increased mortality from cardiovascular diseases. However, the underlying mechanisms are still not clear. We hypothesised that PM, when translocated after inhalation, could affect vascular smooth muscle function. Therefore, total suspended particulate matter (TSP) was sampled and investigated for its ability to affect aortic muscle contraction. Both TSP and TSP supernatant (TSP-sup) induced a concentration-dependent relaxation of phenylephrine (PE) precontracted aortic rings. Relaxation induced by 100 microg/ml TSP was 51.5 +/- 3.1% of total contraction. At 60 and 100 microg/ml, relaxation induced by TSP was significantly higher compared to TSP-sup. Ultrafine TiO2, used as a model to investigate the role of ultrafine particles, did not show an effect. Soluble iron, present in TSP suspensions, seems not to be involved, as chelating with deferoxamine did not affect TSP-induced relaxation. However, TSP effects were inhibited by Trolox, suggesting a role of oxidants. Nudation of aortic rings showed that effects of TSP were only partly endothelium-dependent, while preincubation with L-NAME increased TSP-induced relaxation. From these data, we conclude that both the particle core and soluble components of TSP can affect the smooth muscle function, leading to changes in the vascular contractile response. PMID- 11476160 TI - The effect of hexafluorocyclobutene on rat bronchoalveolar lavage fluid surfactant phospholipids and alveolar type II cells. AB - Hexafluorocyclobutene (HFCB), a reactive organohalogen gas, causes overwhelming pulmonary oedema. We investigated its effect on the rat lung surfactant system, comparing its action on type II pneumocytes with air-exposed rats. The inflammatory cell population and protein content of bronchoalveolar lavage fluid was analysed following exposure to air or HFCB (LCt30). Six rat lung phospholipids were measured by high-performance liquid chromatography, following solid phase extraction (SPE) from lavage fluid. Transmission electron microscopy (TEM) was used to visualise effects on alveolar type II cell ultrastructure. HFCB caused changes in cell populations and increased lavage fluid protein compared to controls, suggesting a permeability oedema. Changes in the total amount and percentage composition (sustained decrease in phosphatidylglycerol and phosphatidylcholine) of surfactant phospholipids also occurred. TEM observations indicated no direct ultrastructural damage to the type II cells, but showed initial, rapid release of surfactant into the alveolar space. HFCB altered the surfactant system in a manner similar to that shown following another reactive organohalogen gas, perfluoroisobutene (PFIB), but differently to that after phosgene. These differences suggest different mechanisms of action even though pulmonary oedema is the final injury for all gases. Better knowledge of the mechanisms involved will improve prospects for prophylactic/therapeutic intervention. PMID- 11476161 TI - Poisoning with Boophane disticha: a forensic case. AB - Scales from the bulb are traditionally used as wound dressing after circumcision and as general wound dressing. Concoctions of the bulb taken orally cause sedation, analgesia, visual hallucinations, irrational behaviour, coma or death. A man ingested 150 ml of a concoction to see who placed a spell on him. He started to hallucinate, thinking that somebody was attacking him. He pulled his gun and fired shots randomly, killing one person and injuring others. A gas chromatograph/mass spectrometer was used to analyze a sample of the concoction. The sample contained buphandrin, buphanine and crinamidine (alkaloids) and eugenol. Buphanine has a pharmacological action similar to that of hyoscine and, when ingested in toxic quantities, leads to excitement, agitation, hallucinations and coma. Eugenol is a volatile oil with analgesic properties. Although itcould not be proved that the concoction was only from Boophane disticha, the components were similar to those found in Amaryllidaceae to which Boophane belongs. The man's behaviour could be ascribed to the ingestion of compounds found in B. disticha. PMID- 11476162 TI - Vulnerability of positron emission tomography radiotracers to endogenous competition. New insights. AB - PET and SPECT neuroreceptor imaging techniques combined with pharmacological challenges have been introduced to measure acute fluctuations of synaptic dopamine (DA) concentrations in the living human brain. Changes in the in vivo binding of radioligands following manipulation of transmitter levels are generally believed to be driven by binding competition between the radioligand and neurotransmitter. This imaging modalityhas been very successful in the study of DA transmission at D2 receptors. Yet, the extension of this technique to the study of other neurotransmitter systems has proven difficult. This paper reviews recent evidencesuggesting that simple binding competition might not be the only phenomenon regulating transmitter-radioligand interactions in vivo, and examines emerging data indicating that receptor trafficking might also be involved. A better understanding of the mechanisms underlying these interactions should facilitate the development of PET and SPECT radiotracers suitable for the reporting of synaptic transmitter levels. PMID- 11476163 TI - Brain radioligands--state of the art and new trends. AB - Non-invasive radioligand imaging methods for brain receptor studies use either short-lived positron-emitting radionuclides such as 11C and 18F for positron emission tomography (PET) or single photon-emitting radionuclides such as 123I for single photon emission computed tomography (SPECT). PET and SPECT use radioligands which are injected intravenously into experimental animals, human volunteers or patients. The main applications of radioligands in brain research concern human neuropsychopharmacology and the discovery and development of novel drugs to be used in thetherapy of neurological and psychiatric disorders. A basic problem in PET and SPECT brain receptor studies is the lack of useful radioligands with appropriate binding characteristics. Prerequisite criteria need to be satisfied for a radioligand to reveal target binding sites in vivo. This section will discuss these important criteria and also review recent examples in neuroreceptor radioligand development such as selective radioligands for brain monoamine transporters. PMID- 11476164 TI - New trends in peptide receptor radioligands. AB - The high level expression of somatostatin receptors (SSTR) on various tumor cells has provided the molecular basis for successful use of radiolabeled octreotide/lanreotide analogs as tumor tracers in nuclear medicine. The vast majority of human tumors seem to overexpress the one or the other of five distinct hSSTR sub-type receptors. Whereas neuroendocrine tumors frequently overexpress hSSTR2, intestinal adenocarcinomas seem to over-express more often hSSTR3 or hSSTR4, or both of these hSSTR. In contrast to 111In-DTPA-DPhe1 octreotide (OCTREOSCAN) which binds to hSSTR2 and 5 with high affinity (Kd 0.1-5 nM), to hSSTR3 with moderate affinity (Kd 10-100 nM) and does not bind to hSSTR1 and hSSTR4, 111In/90Y-DOTA-lanreotide was found to bind to hSSTR2, 3, 4, and 5 with high affinity, and to hSSTR1 with lower affinity (Kd 200 nM). Based on its unique hSSTR binding profile, 111In-DOTA-lanreotide was suggested to be a potential radioligand for tumor diagnosis, and 90Y-DOTA-lanreotide suitable for receptor-mediated radionuclide therapy. As opposed to 111In-DTPA-DPhe1-octreotide and 111In-DOTA-DPhe1-Tyr3-octreotide, discrepancies in the scintigraphic results were seen in about one third of (neuroendocrine) tumor patients concerning both the tumor uptake as well as detection of tumor lesions. On a molecular level, these discrepancies seem to be based on a "higher" high-affinity binding of 111In DOTA-DPhe1-Tyr3-octreotide to hSSTR2. Other somatostatin analogs with divergent affinity to the five known hSSTR subtype receptors have also found their way into the clinics, including 99mTc-HYNIC-octreotide or 99mTc-depreotide (NEOSPECT; NEOTECT). Most of the imaging results are reported for neuroendocrine tumors (octreotide analogs) or non-small cell lung cancer (99mTc-depreotide), indicating high diagnostic capability of this type of receptor tracers. Consequently to their use as receptor imaging agents, hSSTR recognizing radioligands have also been implemented for experimental receptor-targeted radionuclide therapy. The study "MAURITIUS" (MulticenterAnalysis of a Universal Receptor Imaging and Treatment Initiative, a eUropean Study), a Phase IIa study, showed in patients with a calculated tumor dose >10 Gy/GBq 90Y-DOTA-lanreotide, the proof-of principle for treating tumor patients with receptor imaging agents. Overall treatment results in >60 patients indicated stable tumor disease in roughly 35% of patients and regressive disease in 15% of tumor patients with different tumor entities. No acute or chronic severe hematological toxicity, change in renal or liver function parameters due to 90Y-DOTA-lanreotide, was reported. 90In-DOTA DPhe1-Tyr3-octreotide may show a higher tumor uptake in neuroendocrine tumor lesions and may therefore provide even better treatment results in tumor patients, but there is only limited excess to long-term and survival data at present. Besides newer approaches and recent developments of 188Re-labeled radioligands no clinical results on the treatment response is available yet. In conclusion, several radioligands have been implemented on the basis of peptide receptor recognition throughout the last decade. A plentitude of preclinical data and clinical studies confirm "proof-of-principle" for their use in diagnosis as well as therapy of cancer patients. However, an optimal radiopeptide formulation does not yet exist for receptor-targeted radionuclide therapy. PMID- 11476165 TI - Modern trends in radioimmunotherapy of cancer: pretargeting strategies for the treatment of ovarian cancer. AB - A review of published data on some of the problems associated in treating cancer using radioimmunotherapy is presented. Potential improvements for this type of therapy using pretargeting strategies are discussed and preliminary results on a novel multistep regimen to treat human ovarian cancer are presented. A pretargeting strategy using a biotinylated, anti-CA125 monoclonal antibody (MAb) to attract biotinylated long-circulating liposomes to the surface of CA125 expressing ovarian cancer cells, was employed. Confocal laser scanning microscopy and fluorescent labels were used to establish the biodistribution patterns in NIH:OVCAR-3 (CA-125 positive) and SK-OV-3 (CA-125 negative) human ovarian cancer cells. Shedding kinetics of the pretargeted stage were measured using 125I labeled MAbs. No significant internalization of the MAb used in the pretargeting step was observed by 4 hrs. The antibody was gradually internalized starting at 6 hrs, and most of the labelled MAb was detected in cytoplasm by 24 hrs. Shedding and exocytosis of the antigen-MAb complex was not significant for up to 6-hours following administration of the iodinated MAb. Biotinylated liposomes were shown to specifically target the biotinylated MAb/streptavidin complex on the cell surface. We have demonstrated that by a three-step pretargeting approach, biotinylated liposomes can be specifically delivered to cells pretargeted with biotinylated MAb/SAv complex. The slow internalization and shedding properties of the two MAbs are ideal for multistep pretargeting methods. A successful multistep linkage was established with the biotinylated MAb B27.1, streptavidin and biotinylated liposomes to OVCAR-3 cells, but not to SK-OV-3 cells. PMID- 11476166 TI - New concepts in infection/inflammation imaging. AB - Although autologous leukocytes, labelled with 111In or 99mTc, is still considered the "gold standard" nuclear medicine technique to image infection and inflammation, there is a great need for a less cumbersome and less hazardous approach. Over the last few decades the range of radiopharmaceuticals to investigate infectious and non-microbial inflammatory disorders is vastly expanding. Radiolabelled monoclonal antibodies and antibody-fragments, radiolabelled chemotactic peptides and cytokines, and radiolabelled antibiotics are promising new approaches in the field of nuclear medicine. Recently, positron emission tomography (PET) with 18FJDG has been introduced and has been shown to delineate infectious and inflammatory foci with high sensitivity. Here, a survey is presented of the different approaches in use or under investigation. PMID- 11476167 TI - Evaluation of alternative approaches for imaging cellular growth. AB - Uncontrolled growth is a characteristic of malignant tumors. Histochemical techniques to measure tumor growth rate in tissue specimens have proved useful, but are limited because of sampling and the difficulty of following response to therapy. PET imaging offers the opportunity to measure tumor growth non invasively and repeatedly as an early assessment of response to therapy. Measuring cellular growth instead of energy metabolism offers significant advantages in evaluating therapy. The rationale is that a cell's biosynthetic machinery, rather than its fueling process, is more susceptible to cancer therapy. Cytostatic agents may not reduce the quantity of viable tumor; so imaging a change in cellular proliferation may be the only effective way to assess the response to therapy. Radiopharmaceuticals to image growth include labeled amino acids, lipid precursors, and nucleosides. The biochemical characteristic that most uniquely distinguishes successfully treated cancer cells is that they no longer synthesize DNA and no longer divide. Thus imaging with labeled thymidine, which is incorporated into DNA but not into RNA, provides definitive evidence of a cell that is proliferating and, therefore, whether it has responded to treatment. PMID- 11476168 TI - Tumor angiogenesis targeting using imaging agents. AB - The inhibition of tumor induced angiogenesis is an emerging therapeutic strategy in clinical oncology aimed at halting cancer progression by suppressing tumor blood supply. As anti-angiogenic therapy is primarily cytostatic and not cytotoxic, the established criteria for assessing tumor response to chemo- and radiotherapy cannot be applied to anti-angiogenic therapy. Therefore, functional and molecular parameters for imaging of tumor angiogenesis are being intensively studied. Computed tomography, magnetic resonance imaging, ultrasound and scintigraphic techniques can assess changes in vascular permeability and tumor blood flow during anti-angiogenic therapy. Scintigraphic techniques, especially positron emission tomography (PET), may be used to monitor the consequences of anti-angiogenic therapy on tumor cell metabolism, proliferation and apoptosis. The high sensitivity of PET which allows measurements of tracer concentrations in the picomolar range is promising for the visualization of specific molecular targets prior to therapy thus identifying patients most likely benefit from a particular form of anti-angiogenic therapy. PMID- 11476169 TI - PET imaging of hypoxia. AB - Hypoxia in tumors has been related to poor response to conventional therapies. This paper will discuss the methods, both invasive and non-invasive, used to determine hypoxia levels within tumors. PET imaging with two lead compounds 18F fluoromisonidazole (18FMISO) and Cu(II)-diacetyl-bis(N4-methylthiosemicarbazone (Cu-ATSM) and their relative effectiveness in delineating hypoxic regions will be discussed. The advantages of Cu-ATSM-PET over existing imaging agents will be discussed along with its potential application as a direct- and/or surrogate marker for the determination of oncological hypoxia in vivo. PMID- 11476170 TI - Imaging tumors with peptide-based radioligands. AB - Regulatory peptides are small, readily diffusable and potent natural substances with a wide spectrum of receptor-mediated actions in humans. High affinity receptors for these peptides are (over-) expressed in many neoplasms, and these receptors may represent, therefore, new molecular targets for cancer diagnosis and therapy. This review aims to give an overview of the peptide-based radiopharmaceuticals which are presently already commercially available or which are in advanced stages of their clinical testing so that their broader availability is anticipated soon. Physiologically, these peptides bind to and act through G protein-coupled receptors in the cell membrane. Historically, somatostatin analogs are the first class of receptor binding peptides having gained clinical application. 111In-DTPA-[D-Phe1]-octreotide is the first and only radiopeptide which has obtained regulatory approval in Europe and the United States to date. Extensive clinical studies involving several thousands of patients have shown that the major clinical application of somatostatin receptor scintigraphy is the detection and the staging of gastroenteropancreatic neuroendocrine tumors (carcinoids). In these tumors, octreotide scintigraphy is superior to any other staging method. However, its sensitivity and accuracy in other, more frequent neoplasms is limited. Radiolabeled vasoactive intestinal peptide (VIP) has been shown to visualize the majority of gastrointestinal adenocarcinomas, as well as some neuroendocrine tumors, including insulinomas (the latter being often missed by somatostatin receptor scintigraphy). Due to the outstanding diagnostic accuracy of the pentagastrin test in detecting the presence, persistence, or recurrence of medullary thyroid cancer (MTC), we postulated the expression of the corresponding (ie. cholecystokinin [CCK-] -B) receptor type in human MTC. This receptor is also widely expressed on human small cell lung cancer. Indeed, 111In-labeled DTPA derivatives of gastrin showed excellent targeting of CCK-B receptor expressing tissues in animals and patients. A variety of further peptide-based radioligands, e.g. among many others, gastrin releasing peptide/bombesin, neurotensin, substance-P, pan-somatostatin (somatostatin derivatives which bind to all five receptor subtypes) or glucagon like peptide-1 (glp-1) analogs (the latter for the specific detection of insulinomas), is currently under development. Summarizing, radiolabeled regulatory peptides have opened new horizons in nuclear oncology for diagnosis (and potential internal radionuclide therapy). Future work will probably reveal a multitude of novel potentially clinically useful peptide-based radioligands. PMID- 11476171 TI - Radiolabeled bivalent haptens for tumor immunodetection and radioimmunotherapy. AB - The pretargeting technique referred to as the Affinity Enhancement System (AES) uses bispecific antibodies and radiolabeled bivalent haptens that bind cooperatively to target cells in vivo. Experimental and clinical data demonstrate that DTPA bivalent haptens can deliver large radiation doses to tumor cells with high tumor to normal tissue contrast ratios and long activity residence time in tumors. Preliminary clinical results of radioimmunotherapy of medullary thyroid carcinomas and lung cancers look promising. Very encouraging results in biodistribution and radioimmunotherapy experiments in animals have been obtained with new haptens bearing two histamine-hemisuccinate suitable for 131I, 99mTc and 188Re labeling. Targeting isotopes to double antigen positive tumor cells provides a binding enhancement that increases specificity for tumor cells as compared to single antigen targeting on normal cells. This approach may be beneficial for targeting isotopes to B type acute lymphoblastic leukemia and Burkitt lymphoma, as well as others tumors co-expressing two markers of low specificity, and might increase tumor irradiation with minimal irradiation of normal cells. PMID- 11476172 TI - How many ophthalmic plastic surgeons should be trained? PMID- 11476173 TI - Social implications of blepharoptosis and dermatochalasis. AB - PURPOSE: To investigate the social implications of blepharoptosis and dermatochalasis. METHODS: Two hundred ten individuals rated whole-face photographs of a series of patients based on 11 different personal characteristics: intelligence, threat, friendliness, health, trustworthiness, hard work, mental illness, financial success, attractiveness, alcoholism, and happiness. Preoperative and postoperative photographs of both male and female patients with bilateral blepharoptosis and/or dematochalasis were used. The paired t test was used to compare preoperative versus postoperative ratings on the 11 characteristics. RESULTS: The preoperative photographs were rated more negatively than were the postoperative photographs (p < 0.01) on all 11 characteristics for both male and female patients by the 210 study subjects. CONCLUSIONS: Members of our society view individuals with blepharoptosis and dermatochalasis negatively. These social attitudes may lead to unjust bias toward affected patients, and surgical correction probably provides benefits beyond improved visual function. PMID- 11476174 TI - Superomedial lid crease approach to the medial intraconal space: a new technique for access to the optic nerve and central space. AB - PURPOSE: To present clinical and cadaver studies of a new approach to the medial intraconal space. METHODS: We retrospectively review our clinical results by using a new technique to gain access to the medial intraconal space through a superomedial lid crease incision. We also use cadaver dissection studies to compare this new approach with the medial transconjunctival approach, the lateral orbitotomy without bone-flap, and the lateral orbitotomy with bone-flap. RESULTS: Access to the medial intraconal orbital space through the superomedial lid crease incision allowed optic nerve sheath fenestration or biopsy in 18 eyes with few postoperative complications. One case of tonic pupil, one case of transient vertical diplopia, and two cases of transient medial ptosis were seen. Five cavernous hemangiomas were removed from the intraconal space with this approach, with one case of transient vertical diplopia and one case of transient medial ptosis. The cadaver studies showed that when the superomedial lid crease approach is used, the tangent angle with the optic nerve and the incision-to-nerve distance measurements compare favorably with the medial transconjunctival and the lateral orbital approaches. CONCLUSIONS: The superomedial lid crease approach to the medial intraconal space has a number of advantages over the medial transconjunctival and lateral orbital approaches, including ease of dissection, incision-to-nerve distance, and angle of approach to the optic nerve. This technique allows for optic nerve sheath fenestration or tumor removal from the central space with few complications. PMID- 11476175 TI - Effects of hyperbaric oxygen therapy on hydroxyapatite orbital implant vascularization in rabbits. AB - PURPOSE: To deterrmine if hyperbaric oxygen therapy affects the rate of hydroxyapatite orbital implant vascularization in normal rabbit orbits. METHODS: We performed a randomized comparative experimental pilot study involving 6 rabbits. All rabbits were enucleated and implanted with hydroxyapatite orbital spheres. The animals were randomized for enucleation of the right or left eye and for treatment or nontreatment (control) with hyperbaric oxygen. The implants were removed after 3 weeks of treatment and histologically examined for fibrovascular ingrowth, inflammation, and multinucleated giant cells. Each parameter was graded on a numeric scale and analyzed. RESULTS: Hyperbaric oxygen therapy did not increase implant vascularization compared with nontreatment implants. Although treated implants had less central fibrovascular maturity compared with control implants, the difference was not statistically significant (p < 0.055). There was no significant difference in inflammation or the number of multinucleated giant cells between treated and control implants. CONCLUSIONS: In this pilot study, hyperbaric oxygen therapy did not increase hydroxyapatite vascular ingrowth and possibly delayed fibrovascular maturation in normal sockets. Further studies with more subject numbers are needed to confirm these conclusions. The effect of hyperbaric oxygen therapy in vascularly compromised sockets also needs to be determined. PMID- 11476176 TI - 2-Octyl cyanoacrylate tissue adhesive and muscle attachment to porous anophthalmic orbital implants. AB - PURPOSE: To investigate the tolerance and efficacy of a topical bioadhesive for direct muscle attachment to porous orbital motility implants. METHODS: Ten New Zealand White rabbits underwent unilateral enucleation with unwrapped 12-mm spherical porous implants placed into the muscle cone. Four animals received porous polyethylene implants and 6 received bone-derived hydroxyapatite implants. Small amounts of a bioadhesive (2-octyl cyanoacrylate) were applied to the underside of the muscles. Muscles were placed onto the desired location of the implant with pressure for 15 seconds. The conjunctiva was closed with a 6-0 plain gut suture, and the animals were observed for 6 weeks. At 6 weeks, animals were killed and the surgical orbit was exenterated. Orbits and implants were processed for histologic analysis. In addition, tensile strength testing of muscle-implant attachments was performed and compared with various other types of muscle-implant or muscle-sclera attachments. RESULTS: All cases resulted in integration of the muscles without slippage. All unwrapped hydroxyapatite implants (n = 6/6) had small anterior implant exposures without infection, significant inflammation, or exposure. No exposures were noted in porous polyethylene implants (n = 0/4). All implants were filled with fibrovascular tissue by 6 weeks. 2-Octyl cyanoacrylate yielded tensile strengths similar to that of sutured muscle coupling. CONCLUSIONS: 2-Octyl cyanoacrylate is well tolerated in the orbit, inducing minimal inflammation. It effectively attaches muscles to porous implants without hindering permanent integration or vascularization. The exposure rate for unwrapped hydroxyapatite spheres was not caused by the adhesive, as evidenced by the lack of exposures in the porous polyethylene implant group. 2-Octyl cyanoacrylate may be effective in reducing the cost and time for enucleation surgeries when porous motility implants are used. PMID- 11476177 TI - An animal model of lower eyelid spacer grafting with acellular dermis. AB - PURPOSE: Lower eyelid retraction is common in patients with dysthyroid orbitopathy and is seen less commonly in other conditions. Treatment typically requires vertical augmentation of the posterior eyelid lamella with an interpositional graft. Several autologous, homologous, and alloplastic materials have been used. We investigated the gross and microscopic in vivo histology of acellular homologous dermis used as a structural interpositional graft in the lower eyelids of Yucatan minipigs. METHODS: This prospective, experimental study was designed as follows: Surgery was performed on the left lower eyelid of 4 Yucatan minipigs in accordance with the Massachusetts General Hospital Subcommittee on Research Animal Care guidelines. A 4 x 20-mm strip of acellular porcine dermis was prepared and sutured into place between the tarsus and conjunctiva/lower lid retractors. Tissues were harvested at 6 and 12 weeks and were evaluated histologically. RESULTS: Gross examination revealed vertical elongation of the palpebral conjunctival surface. Histologic evaluation was performed with hematoxylin and eosin and Verhoeff elastin stains. Serial sections demonstrated incorporation of grafted dermal matrix into native tissues without evidence of inflammation. Definitive differentiation between grafted dermis and native dermis was not possible with light microscopy. Portions of the graft appeared to be epithelialized, whereas other histologic regions showed poorly adhesive epithelium. Inflammatory infiltrates including lymphocytes and polymorphonuclear leukocytes were not seen. No epithelioid or giant cells were identified in the sections examined. CONCLUSIONS: Acellular homologous dermis produced no measurable untoward effects when implanted as an interpositional graft in the lower eyelid of Yucatan minipigs. On the basis of its material characteristics, it appears to be suitable for this purpose: It is readily available, easily stored, easy to manipulate, and produces minimal inflammation. Long-term persistence, early postoperative histology, and clinical eyelid elevation in humans remain to be evaluated. PMID- 11476178 TI - Differential section of the seventh nerve as a tertiary procedure for the treatment of benign essential blepharospasm. AB - PURPOSE: To examine the efficacy of differential section of the seventh nerve in treatment of patients with blepharospasm refractory to botulinum toxin and eyelid protractor myectomy. METHODS: A retrospective noncomparative interventional case series consisting of a cohort of 228 patients with benign essential blepharospasm followed from 1987 to 1997 in a university ophthalmic plastic surgery referral practice. Patients were treated with botulinum toxin injections, eyelid protractor myectomy, and differential section of the seventh nerve in stepwise fashion as needed for symptomatic control. RESULTS: Thirty-four patients (15% of total) underwent eyelid protractor myectomy during this period. Eyelid protractor myectomy failed to control blepharospasm in 7 (21%) of these 34 patients, who then underwent differential section of the seventh nerve an average of 2 years after myectomy. Patients were followed up for an average of 36 months, with a success rate of 42% (3 of 7). The remaining 4 patients had repeat differential section of the seventh nerve with a 50% success rate, which brought the overall success rate from differential section of the seventh nerve to 71%. Lower eyelid ectropion requiring surgical repair complicated 27% of differential section of the seventh nerve procedures. CONCLUSIONS: Differential section of the seventh nerve is a reasonable alternative in the treatment of patients who have persistent disability despite treatment with botulinum toxin injections and eyelid protractor myectomy. PMID- 11476179 TI - Involutional entropion repair with fornix sutures and lateral tarsal strip procedure. AB - PURPOSE: To evaluate the long-terrm effectiveness of fornix suture placement combined with a lateral tarsal strip procedure in correcting involutional entropion. Published reports regarding various surgical techniques and results are reviewed. METHODS: This retrospective study reviewed 119 patients with involutional lower eyelid entropion who underwent surgical repair between January 1987 and May 1999 at the Bascom Palmer Eye Institute. Exclusion criteria included follow-up duration of less than 6 months, previous lower eyelid blepharoplasty, previous conjunctival surgery other than chalazion removal, or cicatricial entropion. The three surgical subsets were (1) combined lateral tarsal strip and fornix sutures: (2) fornix sutures alone; and (3) lateral tarsal strip procedure alone. The chart review was complemented by a telephone questionnaire to assess the long-term clinical outcome, complications, and patient satisfaction. RESULTS: One hundred fifty-two eyelids in 119 patients were included. One hundred twenty five eyelids had combined surgery (lateral tarsal strip with fornix sutures), 9 eyelids had only fornix suture repair, and 18 eyelids had repair with only the lateral tarsal strip procedure. The recurrence rate in these three surgical subsets was 1.6%, 33%, and 22%, respectively, with average follow-up of 36 months. One case of incisional cellulitis was encountered. Postoperative ectropion was not seen in the group having the combined lateral tarsal strip and fornix suture procedure. CONCLUSIONS: Suture advancement of the lower eyelid retractors in conjunction with a lateral tarsal strip procedure is a simple, quick, physiologic, and effective approach in achieving long-lasting correction for involutional entropion. PMID- 11476180 TI - Sinonasal undifferentiated carcinoma with orbital invasion: report of three cases. AB - PURPOSE: To report three patients with sinonasal undifferentiated carcinoma (SNUC) that invaded the orbit. METHODS: Retrospective small case series. The clinical, radiographic, and pathologic features of three patients with SNUC were reviewed. RESULTS: Three patients with SNUC that invaded the orbit were evaluated. A biopsy was performed on the tumors, which were composed of small, hyperchromatic cells with numerous mitoses and areas of necrosis. Immunohistochemical staining was positive for cytokeratins AE1.3, epithelial membrane antigen, and neuron-specific enolase in all three tumors. Electron microscopic examination showed absence of neurosecretory granules and presence of basement membrane production. Two patients were treated with surgical resection and postoperative chemotherapy and/or radiation. One patient was treated with preoperative radiation and chemotherapy. CONCLUSIONS: Sinonasal undifferentiated carcinoma is a high-grade tumor that arises in the nasal and paranasal sinuses and may invade the orbit. SNUC should be distinguished from other small, round, blue cell tumors, in particular, esthesioneuroblastoma. PMID- 11476181 TI - Multiple recurrences in malignant peripheral nerve sheath tumor of the orbit: a case report and a review of the literature. AB - PURPOSE: To report the onset of malignant peripheral nerve sheath tumor of the orbit 8 years after irradiation in a patient with neurofibromatosis type-1. METHODS: Case report of a young man with neurofibromatosis type-1 who received irradiation for presumed bilateral optic nerve and chiasmal gliomas and in whom a malignant peripheral nerve sheath tumor later developed. Exenteration with extirpation of the entire contents of the orbit was performed 6 times. RESULTS: Complete recurrence of the tumor occurred after each surgical procedure until the patient died of malignancy. CONCLUSIONS: Our case underscores the risk of irradiation, especially in children with neurofibromatosis type-1, and emphasizes that radiotherapy should never be given as an empirical therapy. The authors believe that irradiation and neurofibromatosis type-1 may, in combination, pose a significant risk for the development of malignancies. Clear-cut indications and a precise tissue diagnosis are desirable before the initiation of radiotherapy, particularly in the pediatric population. We recommend that if irradiation is necessary in persons with neurofibromatosis type-1, regular follow-up is imperative. In view of the hostile nature of malignant peripheral nerve sheath tumor, early aggressive treatment appears to be the only viable alternative at present. PMID- 11476182 TI - Primary Ewing sarcoma of the orbit. AB - PURPOSE: To report the clinicopathologic features of a 17-year-old patient with primary Ewing sarcoma of the orbit. METHODS: The patient was evaluated clinically before surgery with computed tomography scans of the orbit. After surgery, the patient was staged with computed tomography scans and bone scan and was treated with systemic chemotherapy and radiation therapy to the orbit. The orbital biopsy was evaluated with conventional light microscopy and immunohistochemistry. RESULTS: Clinical evaluation revealed proptosis and limited upgaze. Computed tomography scans disclosed a mass involving the superior orbit, anterior cranial fossa, and temporal fossa. Microscopic examination revealed small, poorly differentiated cells with medium-sized nuclei containing finely granular chromatin and small nucleoli. The cytoplasmic borders of the cells were indistinct. A PAS stain revealed modest glycogen in many of the tumor cells. The tumor stained positive for O-13 and vimentin and was negative for neural, skeletal, and lymphoid cell markers. Computed tomography scan, bone scan, and blood chemistries revealed no other site of involvement. After treatment, the clinical symptoms and signs resolved, and there has been no evidence of residual orbital tumor or metastasis. CONCLUSIONS: Primary Ewing sarcoma of the orbit should be considered in the differential diagnosis of children or young adults with proptosis, diplopia, or periorbital swelling. Immunohistochemistry is essential to distinguish Ewing sarcoma from other small round cell tumors. PMID- 11476183 TI - Dextromethorphan affects cocaine-mediated behavioral pattern in parallel with a long-lasting Fos-related antigen-immunoreactivity. AB - In order to understand the underlying mechanisms responsible for the behaviors mediated by dextromethorphan (DM), we examined the effects of DM on locomotor activity and locomotor patterns in mice, and Fos-related antigen immunoreactivity (FRA-IR) of mouse brain following repeated administration of cocaine. Combined treatments (30 min prior to each cocaine administration) with DM dose-dependently decreased locomotor activity for high doses of cocaine (20 mg/kg, i.p./day x 7). DM combinations did not significantly affect hyperactivity for 10 mg cocaine/kg, i.p./day x 7. In contrast, combined treatments with DM increased the locomotor activity for 5 mg cocaine/kg, i.p./day x 7. These results were consistent with alterations in marginal activity. Repeated administration with cocaine or DM increased FRA-IR in the nucleus accumbens (NAc) and striatum which lasted for at least 7 days. Our results suggest that DM exhibits biphasic effects on the locomotor stimulation induced by cocaine, and that locomotor activities are in parallel with FRA-IR of the striatal complex. However, the role of FRA-IR regulated by DM or/and cocaine remains to be further determined. PMID- 11476184 TI - Camptothecin suppresses nitric oxide biosynthesis in RAW 264.7 macrophages. AB - Nitric oxide is an important cellular mediator that plays a role in tumor growth and angiogenesis. The present study was conducted to evaluate whether camptothecin (CPT), a topoisomerase I inhibitor, exhibits antitumor activity through regulation the inducible nitric oxide synthase (iNOS) biosynthesis pathway. Experiment was performed on RAW 264.7 cells, a transformed macrophage like cell line, stimulated with lipopolysaccharide (LPS) plus interferon-gamma (IFN-gamma). Incubation of RAW264.7 cells with CPT (0.1 to 10 microM) inhibited the LPS/IFN-gamma-induced nitrite accumulation in a concentration-dependent manner with an IC50 value of 0.59+/-0.07 microM. Treatment of cells with concentrations of CPT (< or =3 microM) that are not growth inhibitory or cytotoxic strongly inhibited their ability to express iNOS mRNA and iNOS protein, however, without a direct regulatory effect on iNOS activity. Time course analysis also revealed that CPT acted in a fashion similar to the transcription inhibitor actinomycin-D. Thus, the suppressant effects of CPT on LPS/IFN-gamma stimulated NO production seemed to be mediated probably through inhibition of iNOS gene transcription. From this observation we propose that inhibition of NO biosynthesis by CPT may underlie, at least in part, the efficacy of this antitumor agent. PMID- 11476185 TI - Protective effect of polysaccharides-enriched fraction from Angelica sinensis on hepatic injury. AB - A polysaccharides-enriched fraction from the root of Angelica sinensis, which is known for its antiulcer action on the gastrointestinal tract, was isolated and studied for its hepato-protective effect in rodents. Intra-gastric administration of Angelica sinensis polysaccharides-enriched fraction (AP) at the doses of 50 or 75 mg/kg dose-dependently prevented liver toxicity induced by acetaminophen in mice but did not affect the serum acetaminophen concentration. It normalized the rises of serum alanine transferase (ALT) and hepatic nitric oxide synthase (NOS) activities and the decrease of glutathione level in the liver. It also reduced the hepatic malondialdehyde (MDA) concentration. The protective effect was less evident in the carbon tetrachloride (CCl4)-treated animals including mice and rats. In the rat the elevated serum ALT level was unaffected though the MDA level was similarly reduced by the higher dose of AP. In these animals, CCl4 increased the hepatic glutathione level instead while the NOS activity remained unchanged. These findings suggest that the pathogenic mechanisms of both acetaminophen and CCl4 are different. AP is more effective in the protection against liver damage induced by acetaminophen, which is associated with the glutathione depletion and nitric oxide synthase activation in the liver. PMID- 11476186 TI - Differential abnormalities in plasma 5-HIAA and platelet serotonin concentrations in violent suicide attempters: relationships with impulsivity and depression. AB - Brain serotonergic systems may participate in the regulation of mood, impulsivity and aggressive behavior. Because some monoaminergic mechanisms seem to be similar in the central nervous system and peripheral tissues, we tested whether serotonergic or dopaminergic biochemical parameters in peripheral venous blood are related or not to violent suicide behavior.We simultaneously studied plasma serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA) and platelet 5-HT content in patients within 3 days following a violent suicide attempt and in matched healthy controls. We examined their relationship with depression and impulsivity. Twenty seven drug-free suicide attempters and controls were included. Plasma 5-HIAA and platelet 5-HT concentrations were lower in suicide attempters than in controls. Fifteen patients were classified as impulsive (I), including all patients suffering from personality disorder and alcohol abuse, and 12 as non impulsive (NI), mostly melancholics. MADRS scores were similar in both I and NI suicide attempters. When controlling for age, plasma 5-HIAA was lower in I than in NI suicide attempters or controls; these findings are similar to those we observed recently with CSF 5-HIAA in I and NI violent suicide attempters. Contrarily, platelet 5-HT levels were lower in NI than in I patients or controls. Plasma HVA was not associated with suicide behavior. Plasma 5-HIAA concentration was inversely associated with the degree of impulsivity and platelet 5-HT with the intensity of depression. This study indicates that each peripheral serotonergic index is specifically related to a distinct clinical feature and shows differential alteration according to the impulsivity group. In I and NI drug-free violent suicide attempters an inverse figure between plasma 5-HIAA and platelet 5-HT data was observed indicating a non parallelism between these two peripheral variables. Further prospective studies are needed to investigate whether these peripheral serotonergic parameters may be used as helpful early predictors of violent suicide behavior. PMID- 11476187 TI - Some particularities of sodium nitroprusside stimulatory effects on food intake. AB - It was previously shown that sodium nitroprusside (NP) stimulates food (FI) and water (WI) intakes when injected intraperitoneally (i.p.) in male rats deprived for 1-h of both food and water during day time. The present work shows that: 1) intramuscular NP increased only water intake; 2) when NP was i.p. injected simultaneously without 1-h deprivation it slightly reduced both intakes in the first 30 min but stimulated significantly FI between 30 and 60 min, and 3) it increased significantly FI even in absence of available water, while WI in absence of food was practically not affected. In conclusion NP effect on FI does originate mainly in the splanchnic area, it requires a latency of about 30 min, and stimulation of FI by NP triggers an increase in WI and not the other way around. PMID- 11476188 TI - Pharmacological evaluation of (+)-2- [123I]A-69024: a radioligand for in vivo studies of dopamine D1 receptors. AB - (+)-2-[123I] A-69024, [(+)-1-(2-[123I] iodo-4,5-dimethoxy-benzyl)-7-hydroxy-6 methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline], is a specific and enantioselective dopamine D1 receptor radioligand. The in vivo biodistribution of this radioligand in rats showed high brain uptake and a distribution consistent with the density of dopamine D1 receptors. Highest uptake was observed in the striatum (0.65 %ID/g) at 5 min followed by clearance. As a measure of specificity the striatum/cerebellar ratio reached a maximum of 3.9 at 30 min post-injection. Radioactivity in the striatum was reduced by 68% by pre-administration of the D1 antagonist SCH 23390. Pre-administration of other dopamine binding drugs, spiperone (D2), 7-OH-DPAT (D3), and clozapine (D4) displayed no inhibitory effect on (+)-2-[123I]A-69024 accumulation in any brain region. Ketanserin (5-HT2/5 HT2C) and haloperidol (D2 receptor antagonist/sigma receptor ligand) also displayed no inhibitory effect in any brain region studied. With the pharmacologically inactive enantiomer, (-)-2-[123I]A-69024, the brain uptake was determined to be non-specific since a striatum/cerebellar ratio of near 1 was observed throughout the time course of the experiment. (+)-2-[123I]A-69024 displays enantioselectivity for dopamine D1 receptors and may deserve further investigation as a possible SPECT radioligand. PMID- 11476189 TI - Induction of cholesteryl ester transfer protein in adipose tissue and plasma of the fructose-fed hamster. AB - Cholesteryl ester transfer protein (CETP) plays a pivotal role in the reverse transport of cholesterol and in the remodeling of circulating lipoproteins. While plasma and adipose tissue levels of CETP are affected by a variety of metabolic conditions, the extent of the effects of dietary factors, other than high cholesterol feeding, are not well understood. To further explore this paradigm, male Golden Syrian hamsters were fed for 4 weeks with a 60%-enriched fructose diet (F) and were compared to a matched group of animals fed with a normal chow diet (N). After feeding for 4 weeks, plasma insulin concentrations were lower in animals fed fructose than in control animals (F: 3.3+/-0.8 vs N: 7.4+/-1.9 ng/mL; p<0.03), but there was no significant difference in plasma glucose concentrations between the two groups (F: 138+/-7 vs N: 148+/-10 mg/dL; p>0.05). Fructose-fed animals showed significant increases in plasma triglyceride (F: 269+/-22 vs N: 165+/-22 mg/dL; p<0.01) and plasma cholesterol (F: 150+/-10 vs N: 113+/-6 mg/dL; p<0.02) concentrations compared with control animals. Total CETP activity and immunoreactive mass were higher in the plasma of fructose-fed animals that in that of controls (F: 1036+/-70 vs N: 826+/-43 pmol/h/mL, p<0.04 and F: 24.5+/-3.1 vs N: 37.5+/-4.3 AU, p<0.02, respectively). Adipose tissue CETP mRNA levels, assessed by the very sensitive ribonuclease protection assay, were 53% higher in fructose-fed animals than in controls (F: 14.1+/-2.0 vs N: 9.2+/-1.0 AU over a rRNA control; p<0.04). Adipose tissue CETP activity and immunoreactive mass also showed a statistically significant increase in the fructose-fed hamsters compared with those fed a normal diet (p<0.04). In conclusion, fructose feeding in Syrian hamsters induces a mixed dyslipidemia. These metabolic changes are accompanied by a significant increase in CETP levels, both in plasma and in adipose tissue. This phenomenon suggests that the increase in the expression of adipose tissue CETP may be caused either by the ambient hypercholesterolemia resulting from fructose feeding or by an attenuation of a possible inhibitory effect of plasma insulin concentrations on the expression of adipose tissue CETP in this feeding paradigm. PMID- 11476190 TI - Vascular smooth muscle cell activation and growth by 4-hydroxynonenal. AB - The present study examines the signal transduction mechanism that is involved in the growth of vascular smooth muscle cells exposed to 4-hydroxynonenal (HNE) in vitro. This aldehyde component of oxidized low-density lipoprotein has been identified in atherosclerotic lesion. Exposure to HNE caused ERK, JNK, and p38 MAP kinase activation as well as the induction of c-fos and c-jun gene expression. AP-1 activity was also significantly induced by HNE treatment. These intracellular activities appear to be the mechanism of HNE-caused mitogenesis. Indeed, HNE induced vascular smooth muscle cell proliferation as determened by Alamar-Blue assay and stimulated DNA synthesis as determined by bromodeoxyuridine incorporation. These observations are consistent with a role of lipid peroxidation products in vascular smooth muscle cell growth in atherogenesis. PMID- 11476191 TI - The roles of IL-4, IL-5 and mast cells in the accumulation of eosinophils during allergic cutaneous late phase reaction in mice. AB - Late phase allergic response has been implicated in the pathogenesis of allergic diseases. In the current study, we investigated the role of IL-4, IL-5 and mast cells in the development of cutaneous late phase reaction (LPR) in mice. Antigenic challenge of ears of ovalbumin (OVA)-immunized BALB/c mice caused a biphasic ear swelling peaking at 1 hr (immediate phase reaction; IPR) and 24 hr (LPR). Ear swelling in LPR was significantly suppressed by the treatment with anti-IL-4 monoclonal antibody (mAb) before antigen challenge. Local eosinophil accumulation during LPR, however, was not inhibited by anti-IL-4 mAb. Moreover, anti-IL-5 mAb had no effect on the swelling response though it significantly suppressed the local accumulation of eosinophils. Interestingly, mast cell deficient mice (WBB6F1-W/Wv) developed LPR without exhibiting IPR, while the magnitude of ear swelling and local eosinophilia was significantly lower than in normal congenic mice (+/+ mice). The present findings show that IL-4 and IL-5 differently regulate the development of LPR, and that IgE-mediated mast cell activation is required for full response. PMID- 11476192 TI - Evidence for inhibition of low density lipoprotein oxidation and cholesterol accumulation by apolipoprotein H (beta2-glycoprotein I). AB - Low density lipoprotein (LDL) oxidation and lipid accumulation are thought to enhance the progression of atherosclerosis. Apolipoprotein H (apoH) has been implicated in the development of human atherosclerosis. However, the roles of apoH in the oxidative modification of LDL and cellular accumulation of lipid constituents remained uncharacterized. In this study, the level of plasma apoH was found to be significantly associated with the oxidative susceptibility of LDL in human subjects. Plasma levels of apoH were positively correlated with the lag time but negatively correlated with LDL oxidation rate in conjugated diene formation. By using a J774 A.1 macrophage culture system, we found that apoH could not only inhibit the formation of conjugated diene and thiobarbituric acid reactive substances, but also reduce the electrophoretic mobility of oxidized LDL. Furthermore, apoH decreased cellular accumulation of cholesterol via a reduction in cholesterol influx and an increase in cholesterol efflux. This is the first demonstration that apoH appears to have "antioxidant"-like effects on LDL oxidation. The results also suggest that apoH can inhibit the translocation of cholesterol from extracellular pools to macrophages, suggesting that apoH may play an important role in the prevention of atherosclerosis. PMID- 11476193 TI - In situ model for studying potassium currents in various growth plate chondrocyte subpopulations. AB - A new in situ model of partially digested growth plate cartilage suitable for patch clamp study of membrane currents of chondrocytes from various differentiation stages was developed. Thin sections of growth plate were enzyme digested to expose intact membranes of chondrocytes previously covered by extracellular matrix. This treatment dramatically increased the success rate of tight-seal formation from virtually 0% up to 40%. Whole-cell patch clamp recording revealed a delayed outward rectifying current as the major macroscopic current in chondrocytes of all differentiation stages. This current was sensitive to tetraethylammonium chloride and reversed polarity at a membrane potential close to the equilibrium potential of K+. Chondrocytes at resting stage expressed a much smaller K+ current than the proliferative and hypertrophic chondrocytes. When the current amplitudes were normalized for the cell membrane area, proliferative cells expressed a significantly higher outward current density. PMID- 11476194 TI - Comparison between the effects of pentobarbital or ketamine/nitrous oxide anesthesia on metabolic and endothelial components of coronary reactive hyperemia. AB - Barbiturates induce reduction of myocardial contractility and metabolism, whereas ketamine exerts a sympathomimetic effect that can mask its direct depressant effect on contractility. However, it is unclear whether barbiturates, which interfere with the cytochrome P-450 pathway, or ketamine, which inhibits nitric oxide synthesis, also alter the responsiveness of the coronary vessels to vasodilator stimuli. We hypothesized that the parameters of coronary reactive hyperemia (CRH), which reflect both the degree of myocardial metabolism and vascular reactivity, could be modified by the type of anesthesia used. In two groups of goats, anesthesia was induced either using ketamine plus nitrous oxide or pentobarbital alone. To record coronary flow an electromagnetic flow-probe was placed around the left circumflex coronary artery. In the ketamine group (n = 14) and in the pentobarbital group (n = 16) CRH was studied using the indices of myocardial metabolism and vascular dilator responsiveness. In the pentobarbital group all of the indices of myocardial metabolism were lower than in the ketamine group (i.e. the excess to debt flow ratio was 2.3+/-0.8 vs. 4.6+/-2.4; p< 0.001). Yet, some indices of vascular responsiveness (time derivative of coronary flow and the peak to basal flow ratio) were not different in the two groups. Moreover, the duration of the reactive hyperemia was shorter in the ketamine than in the pentobarbital group (118+/-47 vs. 153+/-45 s, p<0.05). It is suggested that pentobarbital decreases the indices of CRH related to metabolic activity, whereas ketamine reduces the duration of the hyperemic response, which suggests an impairment of endothelial function. PMID- 11476195 TI - Asthma and hayfever in Aboriginal and non-Aboriginal children living in non remote rural towns. AB - OBJECTIVE: To compare the prevalence and risk factors for wheeze, asthma diagnosis and hayfever in Aboriginal and non-Aboriginal children living in rural towns in Australia. DESIGN AND SETTING: Cross-sectional study in two towns in rural NSW, Australia, 1997. PARTICIPANTS: Primary school children (aged 7-12 years) classified by their parents as being of Aboriginal (n = 158) or of non Aboriginal (n = 1,282) origin. MAIN OUTCOME MEASURES: Atopy measured by skinprick tests and respiratory symptoms measured by parent-completed questionnaire. RESULTS: Aboriginal children were less likely to be atopic (36.2% v 45.6%; 95% CI for the difference, -17.6 to -1.3) and to have hayfever (23.3% v 35.2%; 95% CI for the difference, -19.1 to -4.6) than non-Aboriginal children, but were equally likely to have had wheeze (31.0% v 27.3%) and asthma (39.4% v 39.3%). Among Aboriginal children, having had bronchitis before age two was a strong risk factor for wheeze (adjusted odds ratio (aOR), 9.3; 95% CI, 2.8-30.2) and asthma (aOR, 19.3; 95% CI, 4.7-79.3) and having a parent with hayfever was a strong risk factor for hayfever (aOR, 17.9; 95% CI, 3.5-90.8), but these risk factors were weaker among non-Aboriginal children. CONCLUSIONS: Asthma and wheeze are equally prevalent in Aboriginal and non-Aboriginal children living in the same towns, but appear to have a different aetiology. PMID- 11476196 TI - Appendicectomy in Western Australia: profile and trends, 1981-1997. AB - OBJECTIVE: To measure and describe changes in the incidence of appendicectomy in the population of Western Australia (WA) for 1981-1997. DESIGN: Population-based incidence study using hospital discharge data. SETTING: All hospitals in WA (1981 1997). PATIENTS: All patients who underwent an appendicectomy in WA hospitals. MAIN OUTCOME MEASURES: Changes in the incidence of appendicectomy procedures over time; age-standardised rates and age-sex profiles of four appendicectomy subgroups: (1) acute emergency admission, (2) other emergency admission, (3) incidental appendicectomy and (4) other appendicectomy. RESULTS: From 1981 to 1997, there were 59,749 appendicectomies in WA hospitals. The age-standardised rate of appendicectomy declined by 63% in metropolitan females, by 44% in non metropolitan females, by 41% in metropolitan males and by 21% in non-metropolitan males. The rate of decline was significantly greater in females and in metropolitan patients. From 1988 to 1997, acute emergency admission for appendicectomy was the most common admission status and was more common in males than females (122 v 103 per 100,000 person-years) and in non-metropolitan areas. The rate of incidental appendicectomy was higher among females than males (20 v 7 per 100,000 person-years). From 1988 to 1997, recorded diagnosis coding for appendicitis became more specific, with a marked reduction in the use of the "unspecified" appendicitis code. CONCLUSIONS: The overall incidence of appendicectomy has declined markedly in WA and includes a decline in the practice of incidental appendicectomy. The trend was greatest in the metropolitan hospitals. PMID- 11476197 TI - Depression among Australian adolescents. AB - OBJECTIVES: To examine the prevalence and factors associated with depression in Australian adolescents. DESIGN: A representative, multistage probability sample of Australian households conducted in 1998 (part of the National Survey of Mental Health and Well-being). Adolescents completed self-report questionnaires and parents were interviewed using a lay-administered, structured psychiatric interview and several questionnaires. PARTICIPANTS: 1,490 adolescents aged 13-17 years and their parent or main caregiver. MAIN OUTCOME MEASURES: Prevalence of depression in adolescents, as reported by parents and by adolescents themselves; demographic factors; health-risk behaviours; and rate of use of support services. RESULTS: Of the 1,490 adolescents originally sampled, 150 (10%) did not complete responses to questions on depression and were excluded from the analysis. Seventy of the remaining 1340 adolescents (5.2%; 95% CI, 4.0%-6.4%) met criteria for self reported depression. Agreement between parent- and adolescent-reported depression was poor (kappa=0.27). Adolescent-reported depression was associated with increased suicide plans (odds ratio [OR], 2.83; 95% CI, 1.19-6.70) and attempts (OR, 9.05; 95% CI, 3.49-23.50) in the previous year, use of marijuana 10 or more times in the previous month (OR, 2.88; 95% CI, 1.25-6.64), having conduct disorder (OR, 4.09; 95% CI, 1.23-13.63) and use of school support services (OR, 4.71; 95% CI, 1.82-12.22). Those who used any kind of support service (24/70; 34%) used a mean of 2.9 services (mode, 2; range, 1-5). Three per cent (2/70) of depressed adolescents had been treated with antidepressants. CONCLUSIONS: Depressed adolescents exhibit higher rates of health-risk behaviours and psychosocial impairment than non-depressed adolescents, but only a small number receive appropriate treatment. Staff working in school-based services should be trained to identify adolescents with depression and facilitate referral for treatment. PMID- 11476198 TI - Regional variation in the incidence of end-stage renal disease in Indigenous Australians. AB - OBJECTIVE: To evaluate regional variation in the incidence of end-stage renal disease (ESRD) in Indigenous Australians, and to examine the proximity to ESRD treatment facilities of Indigenous patients. DESIGN: Secondary data review, with collection of primary data regarding patients' place of residence before beginning ESRD treatment. PARTICIPANTS: Indigenous ESRD patients who commenced treatment in Australia during 1993-1998. METHODS: We obtained data from the Australian and New Zealand Dialysis and Transplant Registry regarding 719 Indigenous patients who started ESRD treatment between 1 January 1993 and 31 December 1998. We obtained primary data from the treating renal units to determine the place of residence before beginning renal replacement therapy. We calculated the average annual incidence of ESRD for each of the 36 Aboriginal and Torres Strait Islander Commission regions using population estimates based on the 1996 Census, and calculated standardised incidence ratios with 95% confidence intervals for each region. We compared the number of cases with the treatment facilities available in each region. MAIN OUTCOME MEASURE: Regional standardised ESRD incidence for Indigenous Australians referenced to the total resident population of Australia. RESULTS: Standardised ESRD incidence among Indigenous Australians is highest in remote regions, where it is up to 30 times the national incidence for all Australians. In urban regions the standardised incidence is much lower, but remains significantly higher than the national incidence. Forty eight per cent of Indigenous ESRD patients come from regions without dialysis or transplant facilities and 16.3% from regions with only satellite dialysis facilities. CONCLUSIONS: There is marked regional variation in the incidence of ESRD among Indigenous Australians. Because of the location of treatment centres, there is inequitable access to ESRD treatment services for a significant proportion of Indigenous patients. PMID- 11476199 TI - Paralytic shellfish poisoning: a potential public health problem. AB - Paralytic shellfish poisoning (PSP) is a serious illness in which neurological symptoms predominate. Recovery is usually complete and uncomplicated, but in severe cases there may be respiratory paralysis and death. Most cases follow consumption of bivalve molluscs that have filter-fed on toxic marine microalgae (phytoplankton). Microalgae capable of causing the intoxication have been recorded in all States of Australia, and major toxic blooms have occurred in Tasmania, Victoria and South Australia. PMID- 11476200 TI - Survivor of a stingray injury to the heart. AB - Injuries to the extremities from stingray barbs are not uncommon along the Australian seaboard. Cardiac injuries from stingray barbs are rare, even worldwide, and all but one have been fatal. We report a survivor of a cardiac injury caused by a stingray barb. Penetration of a body cavity by a stingray barb requires early surgical referral and management. PMID- 11476201 TI - EBM and the health of Indigenous Australians. AB - Implementing evidence-based medicine (EBM) in primary healthcare for Indigenous people will usually involve increasing services, particularly those for chronic conditions. As shown by the example of diabetes care, there are significant organisational, educational, economic, cultural and structural barriers to implementing EBM in many Indigenous communities. Many of these barriers could be reduced by better-organised service delivery systems at the community level, greater numbers of Indigenous health professionals and greater advocacy for healthy public policy by health services. There is evidence that delivering evidence-based primary healthcare, particularly for chronic diseases, can improve health outcomes in Indigenous communities. There is a need for more investment in strategies to implement EBM and evidence-based public health in Indigenous settings. PMID- 11476202 TI - EBM in practice: aged care. AB - Clinical trials often exclude older people, and complex interventions are incompletely studied, making it difficult to implement evidence-based medicine in some areas of aged care. There is good-quality evidence to guide the treatment of many of the common clinical problems encountered in older people. Use of outcome measures in trials (eg, quality-adjusted life-years or disability-adjusted life years) that are normally applied to the general population may disadvantage older people, for whom regaining independence is the chief concern. There is a gap between the availability of evidence and its application in routine provision of medical care for older people. PMID- 11476203 TI - Allowing the medical use of cannabis. AB - Cannabis has been advocated as a treatment for nausea, vomiting, wasting, pain and muscle spasm in cancer, HIV/AIDS, and neurological disorders. Such uses are prohibited by law; cannabinoid drugs are not registered for medical use in Australia and a smoked plant product is unlikely to be registered. A New South Wales Working Party has recommended granting exemption from prosecution to patients who are medically certified to have specified medical conditions. This proposal deserves to be considered by other State and Territory governments. PMID- 11476204 TI - Infection, wheezing and Aboriginal children. PMID- 11476205 TI - The Australian contribution towards medical training in Malawi. AB - Malawi opened its only medical school in April 1991, despite opposition from some who believed that the venture was a waste of precious resources. From 1986, selected Malawi students had done preclinical training in Australia, the United Kingdom and South Africa, returning to Malawi for the clinical phase of their training. From 1994, Malawi students have been able to do the whole undergraduate medical course in their own country. Twenty Malawi medical students attended the medical schools of the University of Adelaide and Flinders University in South Australia between 1991 and 1996. All but three of those students are now qualified doctors working in various capacities in healthcare delivery service in Malawi. PMID- 11476206 TI - Inaccurate classification of infant deaths in Australia: a persistent and pervasive problem. PMID- 11476207 TI - Does drinking carrot juice affect cancer of the prostate? PMID- 11476208 TI - Does drinking carrot juice affect cancer of the prostate? PMID- 11476209 TI - Does drinking carrot juice affect cancer of the prostate? PMID- 11476210 TI - Does drinking carrot juice affect cancer of the prostate? PMID- 11476211 TI - Chamomile tea enema anaphylaxis. PMID- 11476212 TI - Percutaneous vertebroplasty: a novel treatment for acute vertebral fractures. PMID- 11476213 TI - Percutaneous vertebroplasty: a novel treatment for acute vertebral fractures. PMID- 11476214 TI - History, heritage and hospitals. PMID- 11476215 TI - Appendicectomy--becoming a rare event? PMID- 11476216 TI - Influence of natural, electrically neutral lipids on the potentiometric responses of cation-selective polymeric membrane electrodes. AB - Ionophore-free ion exchanger electrodes were found to exhibit quite a high selectivity for the creatininium ion; however, measurements in diluted urine samples revealed large emf drifts. Potentiometric, chromatographic, NMR, and mass spectrometric evidence did not reveal any major cationic interfering agents, and anionic interfering agents cannot trivially explain the consistently positive emf drifts. Ultrafiltration of urine samples showed that the interfering agents have molecular weights below 1000 u. The drifts are apparently caused by electrically neutral lipophilic compounds of low molecular weight that are easily extracted into organic phases. Follow-up experiments showed that p-cresol and cholesterol cause no significant emf responses but that coproporphyrin, phosphatidylserine, taurocholic acid, cholic acid, phosphatidylethanolamine, and octanoic acid cause positive emf drifts of the type that was observed with the urine samples. The extent of the responses and the response time depend not only on the specific compound but also on the cation in the sample solution. These results suggest that the emf drifts are due to extraction of such natural lipids into the organic membrane phase where they interact in an ionophore-like fashion with the analyte and interfering ions. Changes in the potentiometric selectivities after contact with natural lipids support this interpretation. The same effect of natural lipids is also expected for ionophore-based electrodes. Indeed, exposure of a valinomycin-based electrode to a methylene chloride extract of urine resulted in a significant reduction of the Na+ discrimination, increasing log Kpot(K,Na) from -3.9 to -3.1. PMID- 11476217 TI - Amplification of amperometric biosensor responses by electrochemical substrate recycling. 3. Theoretical and experimental study of the phenol-polyphenol oxidase system immobilized in Laponite hydrogels and layer-by-layer self-assembled structures. AB - The amperometric response toward phenol of PPO-based rotating disk bioelectrodes is analyzed on the basis of a kinetic model taking into account internal and external mass transport effects and a CEC' electroenzymatic mechanism. Monophenolase activity of PPO catalyses the oxidation of phenol to o-quinone (step C). o-Quinone can then enter an amplification recycling process involving electrochemical reduction (step E) and enzymatic reoxidation (step C': catecholase activity). The rate-limiting steps such as monophenolase activity, catecholase recycling, permeability of the membrane, and activity and accessibility of the catalytic enzyme sites are theoretically considered and experimentally demonstrated for different electrode configurations including PPO immobilized in Laponite hydrogels and layer-by-layer self-assembled multilayers of PPO and poly(diallyldimethylammonium). PMID- 11476218 TI - Renewable amperometric immunosensor for Schistosoma japonium antibody assay. AB - A renewable amperometric immunosensor has been proposed for the determination of Schistosoma japonium antibody (SjAb) in rabbit serum. A paraffin-graphite Schistosoma japonium antigen (SjAg) biocomposite, which needs no additional curing, was directly used to construct the immunosensors. The analytical sample containing the desired SjAb was mixed with SjAb labeled with horseradish peroxidase (HRP) to form the incubation solution for the competitive binding assay. Amperometry was used to determine the amount of HRP fixed on the sensor surface, which was related to the content of desired SjAb. Assay conditions were optimized, including the selection of substrate, the loading of SjAg in the biocomposite, the amount of labeled SjAb in the incubation solution, the incubation time, and the temperature. Using o-aminophenol (o-AP) as a substrate, amperometric detection at -200 mV (vs SCE) resulted in a pseudolinear detection range of about 0.36 to 14 microg/mL, with a detection limit of 0.36 microg/mL. Rabbit serum samples with varying infection degrees were analyzed, and the results demonstrated that the concentration that is detectable in this system meets the demands of clinical analyses. A new surface on the immunosensor for use in another competitive assay can be obtained by removing the original one and polishing the surface. PMID- 11476219 TI - Bioluminescence immunoassay for thyroxine employing genetically engineered mutant aequorins containing unique cysteine residues. AB - Genetically engineered one-to-one conjugates between an analyte and a protein label have been demonstrated to yield assays with better detection limits and performance characteristics than those prepared by conventional chemical conjugation methods. To date, the preparation of these conjugates has been limited to fusion techniques where a peptide analyte is fused in frame to the protein label. To further expand the range of analytes that can be detected by using genetic engineering techniques coupled with bioanalytical methods, we have employed site-directed mutagenesis to prepare one-to-one analyte-label conjugates that include nonpeptidic analytes such as drugs, vitamins, and hormones. Specifically, we have prepared mutants of the photoprotein aequorin containing single cysteine residues suitable for site-specific conjugation. Aequorin is a photoprotein that emits light at 469 nm and has been employed as a highly sensitive bioluminescent label in the development of binding assays for important biomolecules. We have performed polymerase chain reaction-based site-directed mutagenesis on apoaequorin to yield four mutant aequorins containing unique cysteine residues at positions 5, 53, 71, and 84 in the polypeptide chain for the purpose of site-specific conjugation to a model analyte. A maleimide-activated thyroxine was selected as the model analyte and site-specifically conjugated to the mutants through their unique cysteine residues. A heterogeneous assay for thyroxine was then developed by employing the genetically engineered aequorin mutants. PMID- 11476220 TI - Influence of pressure upon coupling pressurized capillary electrochromatography with nuclear magnetic resonance spectroscopy. AB - In this work, the influence of supplementary pressure on the separation efficiency of pressurized capillary electrochromatography (pCEC) was examined. At low pressures of up to 30 bar, which is more than sufficient to prevent bubble formation, no significant loss in separation efficiency is observed. Even at 100 bar, the efficiency of pCEC is still significantly better than without application of an electric field. In addition, analysis times are drastically reduced compared to both capillary electrochromatography (CEC) and capillary HPLC. On the basis of these results, an improved interface for capillary NMR coupling is described and used for the separation and identification of a mixture of unsaturated fatty acid methyl esters. Under these conditions, the analysis time could be shortened by up to a factor of 10 when pCEC is coupled to NMR spectroscopy. PMID- 11476221 TI - Synthesis, characterization, and application of fluorescence sensing lipobeads for intracellular pH measurements. AB - This paper describes the synthesis and characterization of micrometric phospholipid-coated polystyrene particles, named lipobeads, with pH-sensing capability and their application for intracellular pH measurements in murine macrophages. The phospholipids used to coat the particles are labeled with fluorescein (a pH-sensitive dye) and tetramethylrhodamine (a pH-insensitive dye), which serves as a referencing fluorophore for increased accuracy of the pH measurements. The synthesis of the pH-sensing lipobeads is realized by the covalent attachment of the fluorescent phospholipids to the surface of carboxylated polystyrene particles. The pH dynamic range of the sensing particles is between 5.5 and 7.0 with a sensitivity of 0.1 pH unit. The excitation light intensity is reduced to minimize photobleaching of the fluorescein-phospholipid conjugates. The fluorescent lipobeads are used to measure the pH in single macrophages. The lipobeads are ingested by the macrophages and directed to lysosomes, which are the cellular organelles involved in the phagocytosis process. Despite the high lysosomal levels of digestive enzymes and acidity, the absorbed particles remain stable for over 6 h in the cells when they are stored in a phosphate-buffered saline solution at pH 7.4. PMID- 11476222 TI - Multivariate curve resolution of wavelet and Fourier compressed spectra. AB - The multivariate curve resolution method SIMPLe to use Interactive Self-Modeling Mixture Analysis (SIMPLISMA) was applied to Fourier and wavelet compressed ion mobility spectra. The spectra obtained from the SIMPLISMA model were transformed back to their original representation, that is, uncompressed format. SIMPULSMA was able to model the same pure variables for the partial wavelet transform, although for the Fourier and complete wavelet transforms, satisfactory pure variables and models were not obtained. Data were acquired from two samples and two different ion mobility spectrometry (IMS) sensors. The first sample was thermally desorbed sodium gamma-hydroxybutyrate (GHB), and the second sample was a liquid mixture of dicyclohexylamine (DCHA) and diethylmethylphosphonate (DEMP). The spectra were compressed to 6.3% of their original size. SIMPLISMA was applied to the compressed data in the Fourier and wavelet domains. An alternative method of normalizing SIMPLISMA spectra was devised that removes variation in scale between SIMPLISMA results obtained from uncompressed and compressed data. SIMPLISMA was able to accurately extract the spectral features and concentration profiles directly from daublet compressed IMS data at a compression ratio of 93.7% with root-mean-square errors of reconstruction < 3%. The daublet wavelet filters were selected, because they worked well when compared to coiflet and symmlet. The effects of the daublet filter width and compression ratio were evaluated with respect to reconstruction errors of the data sets and SIMPLISMA spectra. For these experiments, the daublet 14 filter performed well for the two data sets. PMID- 11476223 TI - Fluorescence lifetime imaging with near-field scanning optical microscopy. AB - Near-field scanning optical microscopy (NSOM) is a high-resolution scanning probe technique capable of obtaining simultaneous optical and topographic images with spatial resolution of tens of nanometers. We have integrated time-correlated single-photon counting and NSOM to obtain images of fluorescence lifetimes with high spatial resolution. The technique can be used to measure either full fluorescence lifetime decays at individual spots with a spatial resolution of <100 nm or NSOM fluorescence images using fluorescence lifetime as a contrast mechanism. For imaging, a pulsed Ti:sapphire laser was used for sample excitation and fluorescent photons were time correlated and sorted into two time delay bins. The intensity in these bins can be used to estimate the fluorescence lifetime at each pixel in the image. The technique is demonstrated on thin films of poly(9,9' dioctylfluorene) (PDOF). The fluorescence of PDOF is the results of both inter- and intrapolymer emitting species that can be easily distinguished in the time domain. Fluorescence lifetime imaging with near-field scanning optical microscopy demonstrates how photochemical degradation of the polymer leads to a quenching of short-delay intrachain emission and an increase in the long-delay photons associated with interpolymer emitting species. The images also show how intra- and interpolymer species are uniformly distributed in the films. PMID- 11476224 TI - Matrix-assisted laser desorption/ionization mass spectrometry for locating abasic sites and determining the rates of enzymatic hydrolysis of model oligodeoxynucleotides. AB - A method using a combination of exonuclease enzymatic digestion and matrix assisted laser desorption/ionization (MALDI) mass spectrometry was developed to locate model abasic sites in a series of model 21-base oligodeoxynucleotides in which a nucleobase was replaced by a hydrogen atom. The exonuclease digestion rate, with either snake venom phosphodiesterase (SVP) or bovine spleen phosphodiesterase (BSP), clearly slows as the digestion approaches the abasic sites and stops as it reaches it. An oligodeoxynucleotide containing an abasic site in which OH replaces the nucleobase shows similar results. MALDI mass spectra taken at appropriate times during the course of hydrolysis are the basis for rate measurements, and the kinetics also reveal the location of the abasic site. A mathematical treatment of the time-dependent MALDI data was implemented to obtain rate curves and rate constants for the enzymatic digestion of both modified and unmodified oligodeoxynucleotides. PMID- 11476225 TI - Charge-state-dependent sequence analysis of protonated ubiquitin ions via ion trap tandem mass spectrometry. AB - One of the major factors governing the "top-down" sequence analysis of intact multiply protonated proteins by tandem mass spectrometry is the effect of the precursor ion charge state on the formation of product ions. To more fully understand this effect, electrospray ionization coupled to a quadrupole ion trap mass spectrometer, collision-induced dissociation, and gas-phase ion/ion reactions have been employed to examine the fragmentation of the [M + 12H]12+ to [M + H]+ ions of bovine ubiquitin. At low charge states (+1 to +6), loss of NH3 or H2O from the protonated precursor and directed cleavage at aspartic acid residues was observed. At intermediate charge states, (+7, +8, and +9), extensive nonspecific fragmentation of the protein backbone was observed, with 50% sequence coverage obtained from the [M + 8H]8+ ion alone. At high charge states, (+10, +11, +12), the single dominant channel that was observed was the preferential fragmentation of a single proline residue. These data can be readily explained in terms of the current model for intramolecular proton mobilization, that is, the "mobile proton model", the mechanisms for amide bond dissociation developed for protonated peptides, as well as the structures of the multiply charged ions of ubiquitin in the gas phase, examined by ion mobility and hydrogen/deuterium exchange measurements. PMID- 11476226 TI - A wireless electrochemiluminescence detector applied to direct and indirect detection for electrophoresis on a microfabricated glass device. AB - A novel electrochemiluminescence (ECL) detector is presented in this article. The detector is applied for micellar electrokinetic chromatographic separation of dichlorotris(2,2'-bipyridyl)ruthenium(II) hydrate [Ru-(bpy)] and dichlorotris(1,10-phenanthroline)ruthenium-(II) hydrate [Ru(phen)] on a microfabricated glass device. It consists of a microfabricated "U"-shape floating platinum electrode placed across the separation channel. The legs of the U function respectively as working and counter electrode. The required potential difference for the ECL reaction is generated at the Pt electrode by the electric field available in the separation channel during electrophoretic separation. Initial experiments demonstrate a micellar electrokinetic separation and direct ECL detection of 10(-16) mol of Ru(phen) (10(-6) M) and 4.5 x 10(-16) mol of Ru(bpy) (5 x 10(-6) M). Also, preliminary results show the indirect detection of three amino acids. The high voltage at the location of detection does not interfere with the electrochemistry. PMID- 11476227 TI - Optical sensor array and integrated light source. AB - We report a new, solid-state, integrated optical array sensor platform. The optical sensor array and integrated light source (OSAILS) is demonstrated for quenchometric O2 detection. The OSAILS requires low voltages (3-5 V dc), it is stable (< or = 5% RSD over the course of several hours of continuous operation), it is reproducible and reversible (< or =3% RSD), it exhibits a rapid response time (<30 s), and it provides good detection limits (0.2% O2). The OSAILS opens the door to a number of new optical sensor array strategies. PMID- 11476228 TI - Determination of beta-2 microglobulin levels in plasma using a high-throughput mass spectrometric immunoassay system. AB - A high-throughput mass spectrometric immunoassay (MSIA) system for the analysis of proteins directly from biological fluids is reported. A 96-well-format robotic workstation equipped with antibody-derivatized affinity pipet tips was used for the parallel extraction of specific proteins from samples and subsequent deposition onto 96-well arrayed matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOFMS) targets. Interferences from nonspecifically bound proteins were minimized through choice of appropriate affinity pipet tip derivatization chemistries. Sample preparation for MALDI-TOFMS was enhanced through the use of hydrophobic/hydrophilic contrasting targets, which also presented functionalities found to promote matrix/analyte crystal growth. Automated mass spectrometry was used in the unattended acquisition of data, resulting in an analysis rate of approximately 100 samples/h (biological fluid-->data). The quantitative MSIA of beta2m levels present in human plasma samples is given as illustration. PMID- 11476229 TI - Measurements of protein stability by H/D exchange and matrix-assisted laser desorption/ionization mass spectrometry using picomoles of material. AB - Recently, we reported on a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI)-based technique, termed SUPREX, that can be used to measure the thermodynamic stability of a protein (Ghaemmaghami, S.; Fitzgerald, M. C.; Oas, T. G. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 8296-8301). In the work described here, we report on our efforts to optimize the sensitivity of SUPREX analyses. We describe a new sample handling protocol for SUPREX that involves the use of batch chromatography methods with reversed-phase chromatographic media for the microconcentration and desalting of SUPREX samples. Using ribonuclease A as a model protein system, we demonstrate that our new protocol permits the SUPREX analysis of as little as 10 pmol of protein. This amount of protein is 100-fold less than the amount of material required in our initial SUPREX protocol, and it is 1-2 orders of magnitude less than the amount of material required in conventional spectroscopy-based methods for measuring the thermodynamic stability of a protein. PMID- 11476230 TI - Selective, sensitive, and rapid phosphopeptide identification in enzymatic digests using ESI-FTICR-MS with infrared multiphoton dissociation. AB - Rapid screening for phosphopeptides within complex proteolytic digests involving electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR-MS) in the negative ion mode with infrared multiphoton dissociation (IRMPD) accompanied by improved phosphopeptide sensitivity and selectivity is demonstrated with the tryptic digests of the naturally phosphorylated proteins bovine alpha- and beta-casein. All peptides in a complex proteolytic digest can be examined simultaneously for phosphorylation with a 4-s IR laser pulse at 7-11 W where phosphopeptide signature ions form upon irradiation, as the low energy of activation phosphate moiety cleavage transpires without the dissociation of the unphsophorylated peptide population. The tyrosine phosphorylated peptide HGLDN-pY-R, its nonphosphorylated analogue HGLDNYR, the kinase domain of insulin receptor unphosphorylated TRDIYETDYYRK, monophosphorylated TRDIYED-pY-YRK, and triphosphorylated TRDI-pY-ETD-pY-pY-RK were also used as model peptides in this research. The sensitivity and selectivity of phosphopeptides is shown to greatly improve when the volatile base piperidine is used to adjust the pH of th PMID- 11476231 TI - High-throughput peptide identification from protein digests using data-dependent multiplexed tandem FTICR mass spectrometry coupled with capillary liquid chromatography. AB - Tandem mass spectrometry (MS/MS) plays an important role in the unambiguous identification and structural elucidation of biomolecules. In contrast to conventional MS/MS approaches for protein identification where an individual polypeptide is sequentially selected and dissociated, a multiplexed-MS/MS approach increases throughput by selecting several peptides for simultaneous dissociation using either infrared multiphoton dissociation (IRMPD) or multiple frequency sustained off-resonance irradiation (SORI) collisionally induced dissociation (CID). The high mass measurement accuracy and resolution of FTICR combined with knowledge of peptide dissociation pathways allows the fragments arising from several different parent ions to be assigned. Herein we report the application of multiplexed-MS/MS coupled with on-line separations for the identification of peptides present in complex mixtures (i.e., whole cell lysate digests). Software was developed to enable "on-the-fly" data-dependent peak selection of a subset of polypeptides from each FTICR MS acquisition. In the subsequent MS/MS acquisitions, several coeluting peptides were fragmented simultaneously using either IRMPD or SORI-CID techniques. The utility of this approach has been demonstrated using a bovine serum albumin tryptic digest separated by capillary LC where multiple peptides were readily identified in single MS/MS acquisitions. We also present initial results from multiplexed-MS/MS analysis of a D. radiodurans whole cell digest to illustrate the utility of this approach for high-throughput analysis of a bacterial proteome. PMID- 11476232 TI - A study of the Lewis acid-base interactions of vinylphosphonic acid-modified polybutadiene-coated zirconia. AB - Polybutadiene-coated zirconia (PBD-ZrO2) is very useful for reversed-phase separations under a wide variety of conditions. Its excellent chemical (pH = 1 13) and thermal (up to 150 degrees C) stability distinguish it from silica-based reversed phases. Just as with silica-based phases, zirconia's surface chemistry significantly influences the chromatography of certain classes of analytes. Zirconia's hard Lewis acid sites can be chromatographically problematic. Analytes such as carboxylic acids strongly interact with these sites on PBD-ZrO2 and do not elute. Addition of phosphate or other strong, hard Lewis bases to the eluent brings about elution, but the resulting peak is often tailed and broad. Typically, cationic solutes are more retained in the presence of phosphate or fluoride due to adsorption of the Lewis base additives and the concomitant development of a negative charge on the surface. This Coulombic interaction can be used to optimize selectivity, but the reversed-phase-cation-exchange retention can produce broad peaks with excessive retention. As an alternative to adding Lewis bases to the eluent, we studied the effect of permanently modifying PBD ZrO2 by covalently attaching vinylphosphonic acid (VPA) to PBD which was predeposited in the pores of zirconia. We have investigated the chromatography of acids, bases, and small peptides on VPA-modified PBD-ZrO2 (VPA-PBD-ZrO2) and compared it to PBD-ZrO2. VPA-PBD-ZrO2 is a reversed-cation-exchange phase with properties quite different from PBD-ZrO2. The chemical stability of both phases led us to explore how low-pH (1.5-3), ultralow-pH (0), and high-pH (12) eluents effect the retention properties of these mixed-mode phases. Ultralow-pH eluents effectively separate small peptides on both phases. This approach gives lower retention, without sacrificing resolution, and much higher efficiency for small peptides than previously reported. PMID- 11476233 TI - Effects of pore flow on the separation efficiency in capillary electrochromatography with porous particles. AB - The effect of pore flow on the separation efficiency of capillary electrochromatography (CEC) has been studied using columns packed with particles with different pore sizes. A previously developed model was used to predict the (relative) pore flow velocity in these columns under various experimental conditions. Equations are derived describing the effect of pore flow on peak broadening in CEC. The theory has been compared with practice in the reversed phase CEC separation of various polyaromatic hydrocarbons. It is shown, by theory and experimentally, that the mass-transfer resistance contribution to peak dispersion can be effectively eliminated when using porous particles with a high (> or =50 nm) average pore diameter. Moreover, at high pore-to-interstitial flow ratios the flow inhomogeneity contribution (the A term in the plate height equation) is also shown to decrease. Under optimal conditions, a reduced plate height of 0.3 for the nonretained compound could be obtained. It is argued that fully perfusive porous particles can be a more efficient separation medium in CEC than nonporous particles. PMID- 11476234 TI - Dependence of thermal mismatch broadening on column diameter in high-speed liquid chromatography at elevated temperatures. AB - In this paper, we compare a narrow-bore column (2.1-mm i.d.) to a conventional bore column (4.6 mm i.d.) at elevated temperatures under conditions where thermal mismatch broadening is serious and show that narrow-bore columns offer significant advantages in terms of efficiency and peak shape at higher linear velocities. We conclude that the so-called thermal mismatch broadening effect is largely due to a radial retention factor gradient and not a radial viscosity gradient. The lower volumetric flow rates inherent with the use of narrower columns lead to lower linear velocity in the heater tubing and longer eluent residence times in the heater. Thus, with the same heater tubing at the same column linear velocity, narrow-bore columns give better thermal equilibration between the eluent and the column compared to wider bore columns. This means that high-temperature, ultrafast liquid chromatography no longer requires excessively long preheater tubing to thermally equilibrate the eluent to the column temperature. Consequently, the use of narrow-bore columns at high-temperature improves analysis speed and efficiency over wider bore columns. We also discuss the advantages of using liquid heat-transfer media as compared to air as the heat transfer media. We show that an air bath ought not be used to heat the mobile phase because at high temperature (>80 degrees C) and high column linear velocity (> 1.5 cm/s) the length of tubing needed to heat the mobile phase to column temperature is prohibitively long. Using accurate, empirical heat-transfer correlations, we estimated the length of tubing needed to heat the eluent as a function of the column linear velocity for both air and liquid heat-transfer media. PMID- 11476235 TI - Chemically L-phenylalaninamide-modified monolithic silica column prepared by a sol-gel process for enantioseparation of dansyl amino acids by ligand exchange capillary electrochromatography. AB - A new type of chiral monolithic column was successfully developed for the enantioseparation of dansyl amino acids by ligand exchange-capillary electrochromatography (LE-CEC) in this work. The monolithic column matrix was prepared by a sol-gel process and then chemically modified with the spacer (3 glycidoxypropyl)trimethoxysilane and the chiral selector L-phenylalaninamide. After being conditioned with Cu(II) aqueous solution, the ligand exchange-chiral stationary phase (LE-CSP) possesses positive charges. When the external electric field was applied in CEC, electroosmotic flow (EOF) was generated on the surface of LE-CSP in the direction from the cathode to the anode. The EOF was found to be dependent on the applied electric field strength and the composition of the mobile phase. With the increase of pH of the mobile phase, the EOF showed a tendency to decrease. Scanning electron microscopy showed that the chiral monolithic column has a continuous skeleton and large through-pore structure. The separation efficiency (theoretic plate numbers) for the separation of Dns-DL-Leu reached up to 9.0 x 10(4) plates m(-1) for the D-enantiomer and 6.6 x 10(4) plates m(-1) for the L-enantiomer, by using pH 5.5, acetonitrile/0.50 mM Cu(Ac)2 50 mM NH4Ac (7:3) as mobile phase. The reproducibility and lifetime were satisfactory. CEC was carried out with conventional capillary electrophoresis equipment without pressurizing the ends of the capillary. No bubble was formed during the operation, after degassing the mobile phase and conditioning the column. PMID- 11476236 TI - HPLC determination of cyanuric acid in swimming pool waters using phenyl and confirmatory porous graphitic carbon columns. AB - The chlorinated salts of cyanuric acid have found an important role in recreational swimming pool waters across the United States. Upon application to pool water, they can (1) release disinfectant chlorine or (2) stabilize the free available chlorine by acting as chlorine reservoirs in the form of cyanuric acid, preventing the photolytic destruction of residual chlorine by sunlight. Recommended levels of the cyanuric acid stabilizer are in the 10-100 mg/L concentration range according to the National Swimming Pool Foundation (San Antonio, TX). Two isocratic HPLC methods with UV detection (213 nm) employing phenyl and porous graphitic carbon (PGC) columns and phosphate buffer eluents (pH 6.7 and pH 9.1, respectively) were developed to accurately measure cyanuric acid in swimming pools. The two methods allowed fast separation and detection of the stabilizer in 4 (phenyl) and 8 (PGC) min. Both methods offered practical sensitivities with method detection limits of 0.07 (phenyl) and 0.02 mg/L (PGC). Neither one of the two methods required the use of sample cleanup cartridges. They exhibit chromatograms with excellent baseline stability enabling low-level quantitation. Most important, the PGC column had a useful lifetime of five months and 500 sample analyses/column. Eleven pool water samples were fortified with 4.8 50.0 mg/L stabilizer, and the average recovery was 99.8%. Finally, statistical analysis on the relative precisions of the two methods indicated equivalence at the 0.05 critical level. PMID- 11476237 TI - On-column trace enrichment by sequential frontal and elution electrochromatography. 1. Application to carbamate insecticides. AB - An on-column trace enrichment method for CEC of dilute samples is presented. The method involves on-line preconcentration by frontal electrochromatography under conditions of strong solute binding to the stationary phase followed by a step gradient elution electrochromatography with a mobile phase of high eluting strength. This method is tested with dilute samples of carbamate insecticides using capillary columns of 100-microm i.d. packed with a 5-microm octadecyl silica (ODS) stationary phase. The effectiveness of on-line preconcentration (i.e., zone narrowing) depends on the retention factor, k', of the analyte in the injection solvent as well as in the eluting mobile phase (i.e., the organic solvent content), the applied voltage during sample introduction, and elution and length of the introduced sample plug. Under optimal frontal and elution electrochromatography conditions, a 500-fold sensitivity increase is achieved for carbofuran (a carbamate insecticide) with a UV detector. The method is demonstrated with deionized and tap water samples spiked with carbamate insecticides. PMID- 11476238 TI - Application of ABTS radical cation for selective on-line detection of radical scavengers in HPLC eluates. AB - The radical cation 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate), (ABTS*+) was utilized in an on-line HPLC method for the detection of radical scavengers in complex matrixes. The HPLC-separated analytes react postcolumn with the preformed ABTS*+, and the induced bleaching is detected as a negative peak by an absorbance detector at 734 nm. An optimized instrumental and experimental setup is presented. The method is suitable for both isocratic and gradient HPLC runs using mobile phases containing 100% organic solvent or its solution in water, weak acids, or buffers (pH 3-7.4). The method is sensitive, selective, relatively simple, applicable to compounds of different chemical natures; uses common instruments and inexpensive reagents; and has a time-saving, nonlaborious experimental protocol. It can also be used for quantitative analysis. The method was applied to several pure natural antioxidants and plant extracts. The minimum detectable concentration varied from 0.02 to 0.13 microg/mL, depending on the compound tested. The method can be applied to perform kinetic studies, which is illustrated by determination of Trolox equivalent antioxidant capacities (TEAC) of several known antioxidants in flow injection mode. PMID- 11476239 TI - Simultaneous determination of dissolved gases and moisture in mineral insulating oils by static headspace gas chromatography with helium photoionization pulsed discharge detection. AB - This paper presents the development of a static headspace capillary gas chromatographic method (HS-GC) for simultaneously determining dissolved gases (H2, O2, N2, CO, CO2, CH4, C2H6, C2H4, C2H2, C3H8) and moisture from a unique 15 mL mineral oil sample. A headspace sampler device is used to equilibrate the sample species in a two-phase system under controlled temperature and agitation conditions. A portion of the equilibrated species is then automatically split injected into two chromatographic channels mounted on the same GC for their separation. The hydrocarbons and the lighter gases are separated on the first channel by a GS-Q column coupled with a MolSieve 5-A column via a bypass valve, while the moisture is separated on the second channel using a Stabilwax column. The analytes are detected by using two universal pulsed-discharge helium ionization detectors (PDHID). The performance of the method was established using equilibrated vials containing known amounts of gas mixture, water, and blank oil. The signal is linear over the concentration ranges normally found for samples collected from open-breathing power transformers. Determination sensitivity varies with the nature of the species considered with values as high as 21 500 A x 10(-9) s (microg/ g)(-1) for H2O, 46-216 A x 10(-9) s (microL/L)(-1) for the hydrocarbons and carbon oxides, and as low as 8-21 A x 10(-9) s (microL/L)(-1) for the O2 and N2 permanent gases. The detection limit of the method is between 0.08 and 6 microL/L for the dissolved gases, except for O2, N2, and CO2, where higher values are observed due to air intrusion during sampler operations, and 0.1 microg/g for the dissolved water. Ten consecutive measurements in the low and high levels of the calibration curves have shown a precision better than 12% and 6%, respectively, in all cases. A comparison study between the HS-GC method and the ASTM standard procedures on 31 field samples showed a very good agreement of the results. The advantages of configuring the arrangement with two PDHID over the conventional flame ionization and thermal conductivity detectors were clearly demonstrated. PMID- 11476240 TI - An immunoassay for small analytes with theoretical detection limits. AB - A flow-based immunoassay that uses microspheres as the solid phase accomplished the theoretical limit of detectability achievable with the antibody. An equilibrated mixture of anti-estriol monoclonal antibody and estriol was briefly exposed to a bead pack containing immobilized estriol in a flow cell. A small portion of free antibody was separated rapidly from the mixture by binding it to immobilized hormone, but the antibody-hormone complex was kinetically excluded from binding. This rapid separation prevented shift in the equilibrium of the liquid phase binding. Signals were generated by labeling the separated antibodies on the beads with a Cy5-conjugated antispecies secondary antibody. By labeling after the separation step, perturbing the liquid-phase or solid-phase binding was prevented. This assay allowed the reduction of the concentration of primary antibody by continuously accumulating free antibody onto the beads prior to quantification and, thus, offered ideal conditions to achieve theoretical limits of detectability. The optimum achievable dynamic range of this immunoassay was 4 300 pM. Because the proportion of free anti-estriol antibody in the mixture was controlled by the Kd of the antibody-estriol interaction, when the concentration of the antibody was below the Kd, the smallest detectable estriol concentration approached the theoretical limit of detectability achievable with this antibody. PMID- 11476241 TI - Electrokinetically driven microfluidic chips with surface-modified chambers for heterogeneous immunoassays. AB - This article presents the first example of a microfluidic chip for heterogeneous bioassays using a locally immobilized biospecific layer and operated electrokinetically. The reaction chamber has picoliter dimensions and is integrated into a network of microchannels etched in glass. The high affinity of protein A (PA) for rabbit immunoglobulin G (rIgG) was exploited for chip testing, with PA being immobilized on microchannel walls and fluorescently labeled (Cy5) rIgG serving as sample. It was possible to operate the chip in an immunoaffinity chromatographic manner, using electrokinetically pumped solutions. Concentration of antibody from dilute solution onto the solid phase was demonstrated, with signal gains of approximately 30 possible. A dose-response curve for Cy5-rIgG was obtained for concentrations down to 50 nM, for an incubation time of 200 s. The flexibility of chip layout was demonstrated for competitive immunoassay of rIgG, using both a combined sample/tracer incubation and sequential addition of these solutions. With assay times generally below 5 min for this unoptimized device, the microfluidic approach described shows great potential for many high throughput screening applications. PMID- 11476242 TI - Radiochemical determination of 241Am and Pu(alpha) in environmental materials. AB - Americium-241 and plutonium determinations will become of greater importance over the coming decades as 137Cs and 241Pu decay. The impact of 137Cs on environmental chronology has been great, but its potency is waning as it decays and diffuses. Having 241Am and Pu as unequivocal markers for the 1963 weapon fallout maximum is important for short time scale environmental work, but a fast and reliable procedure is required for their separation. The developed method described here begins by digesting samples using a lithium borate fusion although an aqua regia leachate is also effective in many instances. Isolation of the Am and Pu is then achieved using a combination of extraction chromatography and conventional anion exchange chromatography. The whole procedure has been optimized, validated, and assessed for safety. The straightforwardness of this technique permits the analysis of large numbers of samples and makes 241Am-based techniques for high resolution sediment accumulation rate studies attractive. In addition, the technique can be employed for the sequential measurement of Pu and Am in environmental surveillance programs, potentially reducing analytical costs and turnround times. PMID- 11476243 TI - Effects of the ionic environment, charge, and particle surface chemistry for enhancing a latex homogeneous immunoassay of C-reactive protein. AB - The role of the solution environment for a light-scattering, latex-particle enhanced, homogeneous immunoassay of C-reactive protein (CRP) has been investigated in order to assess and optimize the immunoagglutination response. Latex particles of 50-170-nm sizes were covalently coupled with an IgG polyclonal antibody and subjected to an extensive optimization regime. This consisted of conditions responsible, in different degrees, for the principal attractive/repulsive forces affecting both colloidal stability and the antibody/antigen interaction: particle size, antibody concentration, ionic strength and species, pH, and amino acid chemistry of the particle surface. Careful control of these parameters was found to be necessary to achieve the desired effects of balancing high colloidal stability in the absence of antigen but promoting a rapid, sensitive, and dose-dependent agglutination with pathological serum samples. In addition, the estimation of fundamental properties governing intermolecular interaction (i.e. the "Hamaker" constant and critical coagulation concentration) was attempted to order to investigate a simple, practical means of defining a colloidal/immunoassay system under "real conditions" as well as "real time". It is concluded that because each antibody system is unique, a similar optimization should be performed in diagnostic immunoassays of this type to maximize their clinical utility. PMID- 11476244 TI - Effect of poly(ethylene glycol), tetramethylammonium hydroxide, and other surfactants on enhancing performance in a latex particle immunoassay of C reactive protein. AB - The influence of a variety and combination of both ionic surfactants and different chain lengths of the polyelectrolyte poly(ethylene glycol) (PEG) on the performance characteristics (with particular reference to signal response) of a homogeneous, latex agglutination immunoassay was investigated. The test analyte was human serum C-reactive protein (CRP), and the antibody reagent consisted of a sheep polyclonal anti-CRP IgG fraction covalently coupled to 50-nm-sized latex including a glycine-capped chloromethylstyrene shell. The amount and rate of immunoagglutination was monitored turbidimetrically after sample addition. It was found that 2.5 mmol/L concentrations of the small cationic surfactant tetramethylammonium hydroxide (TMH), when present alone, substantially increased both reaction rates and sensitivity in the lower clinical ranges of CRP concentration when compared to normally used assay conditions containing PEG and the anionic detergent Gafac. The nonspecific binding (NSB) was also found to be unchanged. Evidence is presented that the TMH enhances the actual antibody antigen interaction as opposed to the known effects of other surfactants in immunocomplex dissociation or in maintenance of colloidal stability. We suggest that the enhancement seen with TMH could be an alternative to PEG and may provide a new means of further extending detection limits. The utility of this type of immunoassay technology could therefore be increased whenever clinically required. PMID- 11476245 TI - UV Raman spectral intensities of E. coli and other bacteria excited at 228.9, 244.0, and 248.2 nm. AB - Resonance Raman spectral intensities per average bacterial cell have been measured quantitatively for Gram-negative Escherichia coli, Citrobacter freundii, and Enterobacter aerogenes, as well as Gram-positive Bacillus subtilis and Staphylococcus epidermidis. Spectra have been obtained from cultures in the lag, log, and stationary growth phases excited in turn by 228.9, 244.0, and 248.2 nm light. Although Raman spectral peak positions (cm(-1)) excited by a given wavelength are very similar for all five bacterial species, the organisms are characterized by significantly different spectral intensity values. Intensity changes are associated with growth phase changes in all of the species as well. A comparison of measured with estimated average intensities has been made for spectra of log-phase E. coli. It is possible to compare measured intensities with intensities estimated for log-phase E. coli on the basis of the knowledge of its known average cellular molecular composition. A significant degree of hypochromism is observed in E. coli nucleic acid spectra. In contrast, strong average hyperchromism characterizes all aromatic amino acid peaks belonging to the same E. coli cells. Results suggest that knowledge of spectral intensity values will enhance significantly the capability to identify bacteria by means of their UV resonance Raman spectra. PMID- 11476246 TI - Rational design of a Nile Red/polymer composite film for fluorescence sensing of organophosphonate vapors using hydrogen bond acidic polymers. AB - The solvatochromic dye Nile Red dispersed in selected hydrogen bond acidic polymer matrixes demonstrated strong fluorescence enhancement at the presence of dimethyl methylphosphonate (DMMP) vapors. Two hydrogen bond acidic polymers were examined as dye matrixes, one with fluorinated alcohol groups on a polystyrene backbone (PSFA) and the other with fluorinated bisphenol groups alternating with oligo(dimethylsiloxane) segments (BSP3). The combination of hydrogen bond acidic polymer (a strong sorbent for DMMP) with the solvatochromic dye led to initial depression of the dye fluorescence and a significant red shift in the absorbance and fluorescence spectra. DMMP sorption changed the dye environment and dramatically altered the fluorescence spectrum and intensity, resulting in a strong fluorescence enhancement. It is proposed that this fluorescence enhancement is due to the competition set up between the dye and the sorbed vapor for polymeric hydrogen-bonding sites. The highest responses were obtained with BSP3. DMMP detection has been demonstrated at sub-ppm DMMP concentrations, indicating very low detection limits compared to previous Nile Red/polymer matrix fluorescence vapor sensors. Nile Red/poly(methyl methacrylate) films prepared for comparisons exhibited substantially lower response to DMMP. Rational selection of polymers providing high sorption for DMMP and competition for hydrogen-bonding interactions with Nile Red yielded flourescent films with high sensitivity. PMID- 11476247 TI - A dual-adsorbent preconcentrator for a portable indoor-VOC microsensor system. AB - The development and testing of a miniature dual-adsorbent preconcentrator for a microsensor-based analytical system designed to determine complex volatile organic chemical (VOC) mixtures encountered in indoor working environments at low part-per-billion levels is described. Candidate adsorbents were screened for thermal-desorption bandwidth and breakthrough volume against 20 volatile organic vapors and subsets thereof as a function of several relevant variables. A glass capillary (1.1 mm i.d.) packed with 3.4 mg of Carbopack X and 1.2 mg of Carboxen 1000 provides sufficient capacity for a 1-L dry-air sample containing all 20 vapors at concentrations of 100 ppb as well as providing a composite half-height peak width of <3 s at a desorption temperature of 300 degrees C and a flow rate of 4 mL/min. Required adsorbent masses increase to 7.0 and 1.5 mg, respectively, for the same mixture at component concentrations of 1 ppm. Vapor breakthrough volumes for the Carbopack X are unaffected by humidity from 0 to 100%RH, but those for the Carboxen 1000 are significantly reduced, requiring an additional 0.9 mg of the latter to avoid premature breakthrough at the 100 ppb level. Good peak shapes, efficient chromatographic separation of preconcentrated sample mixture components, and detection limits in the low-parts-per-billion range using an integrated surface-acoustic-wave (SAW) sensor are achieved. Preconcentrator design and operating parameters are considered in terms of the vapor bed residence times and breakthrough volumes in the context of the modified Wheeler equation. PMID- 11476248 TI - Use of linear solvation energy relationships for modeling responses from polymer coated acoustic-wave vapor sensors. AB - The applicability and performance of linear solvation energy relationships (LSERs) as models of responses from polymer-coated acoustic-wave vapor sensors are critically examined. Criteria for the use of these thermodynamic models with thickness-shear-mode resonator (TSMR) and surface-acoustic-wave (SAW) vapor sensors are clarified. Published partition coefficient values derived from gas liquid chromatography (GLC) are found to be consistently lower than those obtained gravimetrically, in accordance with previous reports, suggesting that LSERs based on GLC-derived partition coefficients will not provide accurate estimates of acoustic-wave sensor responses. The development of LSER models directly from polymer-coated TSMR vapor sensor response data is demonstrated and a revised model developed from SAW vapor sensor response data, which takes account of viscoelastic changes in polymeric coating films, is presented and compared to those developed by other methods. PMID- 11476249 TI - Particle characterization and separation by a coupled acoustic-gravity field. AB - A coupled acoustic-gravity field is proposed as a novel external field for particle separation and characterization. When a standing plane ultrasound wave is generated, particles move to the node of the wave along the ultrasound force gradient. If the particles also undergo a sedimentation force, they aggregate at the equilibrium position, where these two forces are balanced. The equilibrium position, which is determined by the density and compressibility of a medium and particles, characterizes the particles. The local ultrasound energy, which is necessary for quantitative discussions, is evaluated by using a standard particle, the physical parameters of which are unambiguously determined; aluminum particles are used in the present study. The local ultrasound energy makes possible the determination of the compressibility of unknown materials. Nonporous particles of inorganic and polymeric materials, the particle sizes of which range from 3 to 100 microm, follow a derived model, suggesting that the local ultrasound energy and a derived model be valid. The proposed external field can be used for separation of particles having different acoustic natures. PMID- 11476250 TI - On-wafer spectrofluorometric method for determination of relative quantum yields of photoacid generation in chemically amplified resists. AB - Chemically amplified resists (CARs) that employ acid catalysts are widely used throughout the semiconductor industry due to the need for high throughput in the lithography process. The quantum yield of the particular photoacid generator (PAG) used to generate a given acid ultimately limits the photospeed of the CAR. Determination of quantum yields of photoacid generation is therefore an important component of resist design. We report the development of an on-wafer spectrofluorometric technique for this purpose. This technique is based on one first reported by Feke et al. (J. Vac. Sci. Technol. 2000, B18, 136- 139), which involves doping the resist formulations containing the candidate PAGs with a fluorescent pH indicator dye, coating one wafer per PAG, patterning the wafers with a dose ramp, and spectroscopically imaging the wafers. The response curve of each PAG is spatially and spectrally encoded in the fluorescence images of each wafer. We investigate the efficacy of coumarin 6, a dye that was introduced as an acid sensor by Pohlers et al. (Chem. Mater. 1997, 9, 3222-3230) for this application. We further apply this technique to the determination of the quantum yield of photoacid generation of four candidate PAGs for prototype 193-nm CARs. This technique is convenient, fast, robust, and nondestructive. PMID- 11476251 TI - Direct ultraviolet spectrophotometric determination of total sulfide and iodide in natural waters. AB - A technique is described that allows the determination of total dissolved sulfide in natural waters using direct ultraviolet detection of the HS- ion. The concentration of bisulfide is determined by measuring absorption from 214 to 300 nm and then deconvolution of the HS- spectra from the complex spectrum of natural fluids. A nonlinear least-squares fitting approach is used for the deconvolution. At a pH near 8, where >95% of total sulfide is present as HS-, the results are indistinguishable from total sulfide measured using the methylene blue method in a wide range of sample types and matrixes including freshwater from groundwater wells, marine hydrothermal vent fluids, and marine sediment porewaters. The method allows simultaneous determination of other UV-absorbing ions, including nitrate, bromide, and iodide, in samples with low total sulfide concentrations. Bisulfide concentrations can be determined in samples with low background absorption, such as well water and hydrothermal fluids, with a detection limit of < 1 microM. The detection limit for bisulfide in sediment porewaters that have a high organic loading, which produces background absorbances of approximately 0.5 A at 260 nm in a 1-cm cuvette, is 5 microM. The only chemical manipulation required is buffering acidic samples to pH > 7 and filtration of particulate-rich samples. PMID- 11476252 TI - Preconcentration and preseparation procedure for platinum determination at trace levels by neutron activation analysis. AB - A procedure that simultaneously allows both preconcentration and preseparation of platinum for its determination by neutron activation analysis at trace levels (<0.1 microg x g(-1)) in environmental related matrixes is developed. The method is mainly based on a two-step chemical procedure: (i) a preirradiation concentration/separation based on a column chromatography separation where the platinum is previously retained and subsequently separated from Au, Ca, Na, Br, and P; (ii) a postirradiation separation/purification of the 199Au at 158.4 keV used for the platinum determination via [198Pt (n,gamma)-->199Pt (beta-)- >199Au]. The method eliminates the following radionuclides with the gamma lines in the region of 158 keV: 47Sc (daughter of Ca) at 159.3 keV; 199Au (produced by double neutron capture onto the natural 197Au) at 158.4 keV; 123m Te at 158.8 keV. In addition, the background activity deriving from 24Na, 32p, and 82Br is drastically reduced. The method was tested by the analysis of certified rock material and then applied for platinum determination in airborne particulate matter. PMID- 11476253 TI - Trace metal loading on water-borne soil and dust particles characterized through the use of split-flow thin-cell fractionation. AB - The fate and transport of metallic pollutants through a watershed are related to the characteristics of undissolved solid particles to which they are bound. Removal of these particles and their associated pollutants via engineered structures such as settling ponds is one goal of stormwater management. Because the particles most often implicated in metal pollution have nominal diameters of <50 microm, Split-flow thin-cell (SPLITT) fractionation was investigated to study the metal loading as a function of particle settling rate. Several diverse particle samples--soil, urban dust, and parking deck sweepings--were fractionated using this technique, and the metal loadings were quantified with inductively coupled plasma-atomic emission spectrometry. Implications for stormwater management are discussed. PMID- 11476254 TI - Design and performance of a universal sheathless capillary electrophoresis to mass spectrometry interface using a split-flow technique. AB - A split-flow capillary electrophoresis electrospray ionization mass spectrometry (CE/ESI-MS) interface is introduced, in which the electrical connection to the CE capillary outlet is achieved by diverting part of the CE buffer out of the capillary through an opening near the capillary outlet. The CE buffer exiting the opening contacts a sheath metal tube which acts as the CE outlet/ESI shared electrode. In cases in which the ESI source uses a metal needle, the voltage contact to the CE buffer is achieved by simply inserting the outlet of the CE capillary, which contains an opening, into the existing ESI needle (thereby greatly simplifying the CE to MS interfacing). As a result of the concentration sensitive nature of ESI, splitting a small percentage of the CE flow has minimal effect on the sensitivity of detection. In addition, because the liquid is flowing through the opening and out of the capillary, there is no dead volume associated with this interface. Moreover, bubble formation due to redox reactions of water at the electrode does not effect CE/ESI-MS performance, because the actual metal/liquid contact occurs outside of the CE capillary. The sensitivity associated with a sheathless CE/MS interface, the ease of fabrication, universality, and lack of any dead volume make this design a superior CE/ESI-MS interface. The performance of this interface is demonstrated by analyses of a peptide standard and a protein digest using a variety of capillary dimensions. PMID- 11476255 TI - On-line incorporation of cloud point extraction to flow injection analysis. AB - The on-line incorporation of cloud point extraction (CPE) to flow injection analysis (FIA) is demonstrated for the first time. The technical difficulties of inducing the cloud point phenomenon, separating the surfactant-rich phase from the aqueous phase, and detecting trace amounts of analyte(s) in the presence of the highly scattering surfactant medium in an on-line FIA system were resolved by the following: (1) mixing the sample solution containing the analyte(s) and CPE surfactant with an appropriate salting-out agent, (2) using a collection column to entrap the analyte-containing surfactant aggregates, and (3) employing the peroxyoxalate chemiluminescence reaction for the sensitive and selective determination of the analyte(s) in the presence of surfactant micelles. The figures of merit for the determination of coproporphyrin in pretreated urine samples were as follows: precision, 1.1-2.2% (RSD); limit of detection, 2.0 microg/L; and the calibration curve was linear from 46 to 2319 (micro/L (r = 0.9996). PMID- 11476256 TI - Analysis of oxidative degradation products of 2,4,6-trichlorophenol treated with air ions. AB - The analysis of oxidative degradation products of 2,4,6-trichlorophenol (TCP) treated with air ions, which are generated by electric discharge, is reported. Due to the complex nature of the degradation products, a combination of different detection techniques was employed to characterize them. The oxidative degradation of TCP is usually dependent on the treating approaches, and in this system, a stepwise degradation, beginning with the formation of a major product 2,6 dichloro-1,4-benzenediol as well as other minor ones (e.g., 3,5-dichlorocatechol) via substitution, is first proposed through a detailed analysis of GC/MS, etc., though some chromogenic quinones can transiently be present. Furthermore, high dechlorination (53%) was observed for TCP after a 60-min treatment, indicating that air ions can serve as an efficient dechlorination means. PMID- 11476257 TI - Effects of high-dose heroin versus morphine in intravenous drug users: a randomised double-blind crossover study. AB - The purpose of this study is to evaluate the effects of high doses of injected opiates as prescribed maintenance in intravenous drugs users. This was accomplished via a randomised double-blind study with crossover at an outpatient clinic in Bern, Switzerland. The subjects were 39 patients with a long history of intravenous opioid use and persistent abuse despite treatment; they were randomly allocated to two groups. Group A was started on controlled injection of graduated doses of morphine up to a satisfying individual dose and was then switched as a double blind to heroin at a randomly determined day between week three and four. Subsequently this group was given heroin for the remaining two to three weeks of the study. Group B was started on heroin and was then switched to morphine in the same manner. Equipotent solutions of 3% morphine and 2% heroin were administered. The main outcome measures were clinical observations, structural interviews and self report of subjective experiences to assess the effects of the drugs. In 16 cases, the study had to be discontinued owing to severe morphine-induced histamine reactions. Thirteen participants in Group B presented these adverse reactions on the day of the switch-over. Full data were thus only obtainable for 17 participants. Average daily doses were 491 mg for heroin and 597 mg for morphine. The findings indicate that heroin significantly produced a lower grade of itching, flushing, urticaria and pain/nausea. A negative correlation between dose and euphoria was observed for both heroin and morphine. The authors concluded that as heroin produces fewer side effects it is the preferred high dose maintenance prescription to morphine. The perceived euphoric effects are limited in both substances. PMID- 11476258 TI - "Have a piss, drink ogogoro, smoke igbo, but don't take gbana"--hard and soft drugs in Nigeria: a critical comparison of official policies and the view on the street. AB - One of the consequences of increased exposure to Western influences has been the spread of drug use among Nigerian young people over the last two decades. There is now a buoyant consumer market for home-grown cannabis, and a smaller market for heroin and cocaine within the country. Under successive military regimes, drug use was either downplayed, or regarded as a law enforcement issue. Little work was carried out to gauge the extent of use, or obtain an understanding of the user constituencies. Government policies are entirely informed by Western models and sadly inadequate in understanding Nigerian drug trends and in devising schemes to meet local needs. This article, by giving voice to Nigerian drug users, is an attempt to dispel some misconceptions about drug use and abuse, and to widen the debate surrounding African drug control. There is a grave danger of the war on drugs being exported to African countries where civil societies are weakest in withstanding the onslaught of government and international agencies. This article is therefore an ethnographic drugs study engaging with crucial development issues. PMID- 11476259 TI - Sticky business: patterns of procurement and misuse of prescription cough syrup in Houston. AB - Reports of recreational codeine cough syrup use have increased in Houston and in the state of Texas. Occasional and polydrug users increasingly have consumed codeine cough syrup (with or without alcohol or other drugs) over the past three years, accounting for a $40 increase in the price of an eight-ounce bottle on the underground economy. News stories regarding syrup abuse and reports of deaths by codeine overdoses suggested the need to explore this emerging drug trend. The investigator conducted a literature search of scientific journals and news media, interviews with community authorities, and guided interviews with 25 adults who reported using codeine cough syrup in the 30 days preceding their interviews. Participants were recruited through snowball sampling; interview transcripts were coded and content analyzed. Polydrug users reported a penchant for codeine syrup because it carries fewer legal consequences, is perceived as "safer" than illegal drugs, and is either free or inexpensive for users with Medicaid or private insurance. Participants reported methods for procuring syrup from physicians and hospital emergency rooms which they consumed or traded for money, goods, or services. Consumption patterns for chronic and occasional users are described. Reported side effects include a drowsy relaxed high, fatigue, loss of coordination, constipation, and urinary retention. PMID- 11476260 TI - Long-term therapy using GHB (sodium gamma hydroxybutyrate) for treatment resistant chronic alcoholics. AB - Thirty-five alcohol-dependent patients according to DSM-IV criteria who also met criteria for treatment resistance were treated with doses of gamma hydroxybutyrate (GHB) ranging between 25 and 100 mg/kg/die in an open one-year study. The results show that no patients discontinued the program during the first month of treatment. Sixty percent of these patients successfully completed the protocol; 11.4% showed complete abstinence (full responder patients); 14.3% strongly reduced their alcohol intake (partial responder patients) and 34.3% of the patients were still under treatment after one year. Forty percent of the patients were nonresponders. The retention rate under treatment of the studied sample was statistically higher than that found during the last treatment of the same subjects. No significant differences were found between full responder and partial responder patients regarding changes in clinical features, alcohol intake or social adjustment. Patients still in treatment after one year significantly differed from nonresponder patients on all the variables investigated. A six times/daily fractionated administration of the GHB dose was the only significant predictor of the retention rate. PMID- 11476261 TI - Gamma hydroxybutyrate (GHB) and gamma butyrolactone (GBL) withdrawal: five case studies. AB - There is little medical information available about gamma-hydroxybutyrate (GHB) or gamma-butyrolactone (GBL) dependence or withdrawal. In this study the authors treated and reviewed multiple cases of GHB and GBL withdrawal in high-dose users. Five patients during nine hospitalizations were treated for GHB or GBL withdrawal. The authors describe a spectrum of GHB or GBL withdrawal from mild to severe and discuss medications used for treatment. They conclude that patients with GHB or GBL withdrawal may present with agitated psychosis, delirium, and autonomic instability. In this sample, relapse to GHB or GBL use occurred soon after treatment of withdrawal. PMID- 11476262 TI - The nonmedical use of ketamine, part two: A review of problem use and dependence. AB - This is the second part of a review of the nonmedical use of ketamine. Part one discussed the history of ketamine, the sought-after effects for which it is taken in a nonmedical context, how these are produced, common adverse effects, the ketamine schizophrenia model and the neurotoxicity issue. Part two reviews what is currently known about problem use of ketamine, ketamine dependence, treatment options and harm minimization issues. Some ketamine users become dependent on the drug in a manner resembling cocaine dependence, with craving and a high tolerance but no evidence of a physiological withdrawal syndrome. The likely mechanisms of this dependence are discussed in terms of what is known about the neurochemistry of ketamine, its psychological effects, and published case histories in both the formal and informal literature. The conclusions are that ketamine dependence is linked with effects that this complex drug has in common with not only cocaine and amphetamine but also with opiates, alcohol and cannabis, as well as the psychological attractions of its distinctive psychedelic properties. PMID- 11476263 TI - The development, reliability, and validity of a clinical rating scale for codependency. AB - This investigation examined the reliability and validity of a rating scale for codependency in substance abuse treatment. The investigators developed an example anchored rating scale to operationalize codependency as substance abuse counselors construe it in practice, and recruited 27 counselors for a counterbalanced multiple-treatment experiment. Counselors were randomly assigned to one of four continuing education workshops for rating-scale training, and asked to evaluate codependency in five videotaped cases. Semistructured case interviews were videotaped with a male and a female from five adult populations to vary the gender and codependency of cases: (1) outpatients in treatment for addiction, (2) outpatient spouses, (3) members of Codependents Anonymous, (4) United States Bureau of Land Management smoke jumpers, and (5) college students majoring in business or economics. To control for gender effects, one workshop presented male cases, one workshop presented female cases, and two workshops presented cases of both genders. To control for order effects, the assignment of videotapes to workshops was randomized to counterbalance the order in which counselors viewed them. The findings suggest that the rating scale yields reliable and valid evaluations of codependency without appreciable gender bias. PMID- 11476264 TI - Walking the tightrope: balancing health and drug use. AB - This article stems from a study on San Francisco drug users and their health care experiences. Two hundred thirty-eight study participants were administered depth interviews and questionnaires to reveal a variety of health concerns and health behaviors. For these analyses, which focused on health practices and risky behaviors, the sample was divided into two groups: marijuana users, and other drug users (including methamphetamine, cocaine, heroin, and crack). The findings indicate that drug users behave similarly to the general population regarding health practices, such as eating regularly, sleeping regularly, taking vitamins, and exercising. One finding of interest was the high-risk sexual behaviors of the young, educated marijuana users. A majority of participants demonstrated strong concerns for their health and well-being, and further encouragement on this front is advised. Future research and directions for interventions are recommended. PMID- 11476265 TI - Assessing the influence of need to inject and drug withdrawal on drug injectors' perceptions of HIV risk behavior. AB - This article aims to assess the influence of the need to inject and drug withdrawal on drug injectors' perceptions of human immunodeficiency virus (HIV) risk behavior inside and outside prison. Complementary qualitative research methods were used with 24 drug injectors in England. It was found that when sterile injecting equipment was unavailable the need to inject and drug withdrawal were important factors on the reported readiness to share injecting equipment. This finding was broadly consistent both outside and inside prison. However, different patterns of responses between these two environments were influenced by the social context in which HIV risk was considered. These perceptions of HIV risk are situationally specific, but the influence of the need for a drug injection and drug withdrawal on HIV risk behavior transcends social settings. Thus, HIV risk reduction strategies should be consistent outside and inside prison. PMID- 11476266 TI - Rapid heroin detoxification using a single high dose of buprenorphine. AB - To test the effect of 32 mg of buprenorphine on the withdrawal process from heroin, 10 street-heroin using subjects were given 32 mg of sublingual buprenorphine, following heroin abstinence of 24 hours. Withdrawal symptoms were monitored during the first few hours, and followed for six days after buprenorphine administration, after which naltrexone (50 mg) was introduced to prevent future heroin use. Nine subjects completed detoxification with negligible withdrawal symptoms and a smooth transition to naltrexone. One subject was excluded from the study due to methadone ingestion prior to experiment. These results strongly suggest that painless detoxification from heroin can be obtained by a single high dose of buprenorphine. PMID- 11476267 TI - HHS releases a clinical guide on the treatment of women with HIV. PMID- 11476268 TI - Federal agencies mark 20th anniversary of first AIDS case. PMID- 11476269 TI - Meperidine or morphine in acute pancreatitis? PMID- 11476270 TI - High blood pressure in pregnancy. PMID- 11476271 TI - NHBPEP report on high blood pressure in pregnancy: a summary for family physicians. AB - The National High Blood Pressure Education Program's Working Group on High Blood Pressure in Pregnancy recently issued a report implicating hypertension as a complication in 6 to 8 percent of pregnancies. Hypertension in pregnancy is related to one of four conditions: (1) chronic hypertension that predates pregnancy; (2) preeclampsia-eclampsia, a serious, systemic syndrome of elevated blood pressure, proteinuria and other findings; (3) chronic hypertension with superimposed preeclampsia; and (4) gestational hypertension, or nonproteinuric hypertension of pregnancy. Edema is no longer a criterion for preeclampsia, and the definition of blood pressure elevation is 140/90 mm Hg or higher. Patients with gestational hypertension have previously unrecognized chronic hypertension, emerging preeclampsia or transient hypertension of pregnancy, an obstetrically benign condition. Because distinguishing among these conditions can be done only in retrospect, clinical management of gestational hypertension consists of repeated evaluations to look for signs of emerging preeclampsia. Women with chronic hypertension should be followed for evidence of fetal growth restriction or superimposed preeclampsia. Management options for chronic hypertension in most women include discontinuing antihypertensive medications during pregnancy, switching to methyldopa or continuing previous antihypertensive therapy. PMID- 11476272 TI - Information from your family doctor. High blood pressure in pregnancy. PMID- 11476273 TI - Radiographic assessment of osteoarthritis. AB - Osteoarthritis is one of the most prevalent and disabling chronic conditions affecting older adults and a significant public health problem among adults of working age. As the bulk of the U.S. population ages, the prevalence of osteoarthritis is expected to rise. Although the incidence of osteoarthritis increases with age, the condition is not a normal part of the aging process. More severe symptoms tend to occur in the radiographically more advanced stage of the disease; however, considerable discrepancy may exist between symptoms and the radiographic stage. Roentgenograms of involved joints may be useful in confirming the diagnosis of osteoarthritis, assessing the severity of the disease, reassuring the patient and excluding other pathologic conditions. The diagnosis of osteoarthritis is based primarily on the history and physical examination, but radiographic findings, including asymmetric joint space narrowing, subchondral sclerosis, osteophyte formation, subluxation and distribution patterns of osteoarthritic changes, can be helpful when the diagnosis is in question. PMID- 11476274 TI - Differential diagnosis of annular lesions. AB - Although most annular lesions will be typical of a dermatophytosis, physicians must consider other possible diagnoses. Tinea corporis can often be diagnosed on the basis of a positive potassium hydroxide examination. Topical and systemic antifungals are usually curative. Pityriasis rosea is characterized by small, fawn-colored lesions distributed along skin cleavage lines. Treatment is symptomatic. Granuloma annulare is characterized by nonscaly, annular plaques with indurated borders, typically on the extremities. One half of cases resolve spontaneously within two years. Sarcoidosis can present as annular, indurated plaques similar in appearance to the lesions of granuloma annulare. Diagnosis is based on histopathology and the involvement of other organ systems. Hansen's disease can mimic tinea corporis by presenting as one or more annular, sometimes scaly, plaques. Urticaria may affect 10 to 20 percent of the population. The annular plaques lack scale and are evanescent. Subacute cutaneous lupus erythematosus can present in an annular form on sun-exposed surfaces or in a papulosquamous form. Erythema annulare centrifugum typically presents as annular patches with trailing scale inside erythematous borders. PMID- 11476275 TI - Can the patient decide? Evaluating patient capacity in practice. AB - Physicians assess the decision-making capacity of their patients at every clinical encounter. Patients with an abrupt change in mental status, who refuse recommended treatment, who consent too hastily to treatment or who have a known risk factor for impaired decision-making should be evaluated more carefully. In addition to performing a mental status examination (along with a physical examination and laboratory evaluation, if needed), four specific abilities should be assessed: the ability to understand information about treatment; the ability to appreciate how that information applies to their situation; the ability to reason with that information; and the ability to make a choice and express it. By using a directed clinical interview or a formal capacity assessment tool, primary care physicians are able to perform these evaluations in most cases. PMID- 11476276 TI - Principles of appropriate antibiotic use: Part I. Acute respiratory tract infections. PMID- 11476277 TI - Reflections on the purinergic hypothesis: the Burnstock Festschrift in the millennial year. AB - Few have made such an impact as Geoffrey Burnstock in their scientific field. As the originator of the purinergic hypothesis, Burnstock has been central to the development of our understanding of the P2 receptor family and of the role of extracellular ATP in cell-to-cell signalling. In this millennial year, Burnstock has been awarded the Queen's medal from The Royal Society and Lifetime Achievement Award from the American Gastroenterology Association. Thus, it was my privilege to join Alan North in organising and producing the Burnstock Festschrift (Purines and the Autonomic Nervous System; from controversy to the clinic, in [J. Auton. Nerv. Syst. Vol. 81 (2000)]) to honour not only Geoffrey Burnstock's successes in this millennial year, but a lifetime of achievements spanning some 40 years in the field of purine signalling. PMID- 11476278 TI - Diminished beta-adrenoceptor-mediated relaxation of femoral arteries from young spontaneously hypertensive rats. AB - A beta-adrenoceptor agonist, norepinephrine (NE)-induced relaxation in the presence of an alpha-adrenoceptor antagonist and indomethacin was investigated in isolated femoral arteries from 5-week-old Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). NE elicited endothelium-dependent and independent relaxation in WKY. In endothelium-intact WKY artery, the NE-induced relaxation was reduced by nitro L-arginine (L-NA) and methylene blue. The residual response to NE in the presence of L-NA was further reduced by tetraethylammonium (TEA). Glibenclamide attenuated the NE-induced, endothelium independent relaxation in WKY. In SHR, on the other hand, the relaxation to NE was solely endothelium-independent, unaffected by a combination of L-NA and TEA and inhibited by glibenclamide. The relaxation in response to NE in SHR was less than that in WKY, regardless of the presence and absence of endothelial cells. When WKY and SHR were treated for 10 days with captopril, the response to NE was increased not only in WKY but also in SHR. The relaxation in captopril-treated SHR consisted of endothelium-dependent and -independent components. The former was attenuated by L-NA and to a greater extent by TEA with L-NA. Sodium nitroprusside- and forskolin-induced, endothelium-independent relaxations in SHR were not significantly different from those in WKY. Captopril did not affect the response to these drugs. The present results indicate that the relaxation to NE is in part mediated by NO and a vasorelaxing factor distinct from NO in WKY but not in SHR. It is suggested that NE-induced, endothelium-independent relaxation in both groups is in part mediated by ATP-sensitive K+ channels. It is also suggested that in SHR, captopril increases the response to NE through increases in endothelial production of NO and the non-NO vasorelaxing factor. PMID- 11476279 TI - Projections of the paratrigeminal nucleus to the ambiguus, rostroventrolateral and lateral reticular nuclei, and the solitary tract. AB - The paratrigerminal nucleus (Pa5), a constituent of the spinal interstitial system, was linked to the pressor effect caused by bradykinin injected in the dorsal lateral medulla of the rat. The nucleus receives primary afferent sensory fibers contained in branches of the trigeminal, glossopharyngeal and vagus nerves. In this investigation connections of the paratrigeminal nucleus to other medullary structures were studied with the use of retrograde and anterograde neuronal tracers. Fluorescent light microscopy analyses of medullary sections of rats injected with the retrograde transport tracer Fluoro-gold in the nucleus of the solitary tract (NTS) or in the pressor area of the rostral ventrolateral medulla (RVLM) revealed labeled neuronal cell bodies in the ipsi- and contralateral Pa5. FluoroGold microinjections in the caudal ventrolateral medulla (CVLM) did not produce fluorescent labeling of Pa5 neurons. Microinjection of the anterograde transport neuronal tracer biocytin in the Pa5 produced bilateral labeling of the solitary tract (sol). rostroventrolateral reticular nucleus (RVL), ambiguus nucleus (Amb), lateral reticular nucleus (LRt) and ipsilateral parabrachial nuclei, but not the contralateral Pa5. Confocal laser microscopy showed fluorescence labeling of fibers and presumptive terminal varicosities in the NTS, RVL, Amb and LRt. The present findings showing the paratrigeminal nucleus interposed between sensory afferent and stuctures associated to cardiovascular and respiratory functions, suggest that the structure may act as a medullary relay nucleus for sensory stimuli directly connecting primary afferents to structures mediating cardiovascular and respiratory reflexes. PMID- 11476280 TI - Effects of ketamine and propofol on autonomic cardiovascular function in chronically instrumented rats. AB - In this study C. we systematically examined the effects of ketamine and propofol at various doses (5-20 mg/kg) on blood pressure, heart rate and renal sympathetic nerve activity in chronically instrumented Wistar rats. We also assessed the effects of these anesthetics on the baroreflex control of heart rate and renal sympathetic nerve activity. Ketamine (10 mg/kg) increased blood pressure by 30.0+/-4.5%, heart rate by 17.7-3.3% and renal sympathetic nerve activity by 38.8+/-14.6%, while propofol (10 mg/kg) decreased blood pressure by 18.9+/-3.5%, heart rate by 5.5+/-2.5% and renal sympathetic nerve activity by 7.5+/-2.1%. These variables showed dose-dependent responses to both agents. Both ketamine and propofol decreased the range and maximum gain of the logistic function curve obtained by relating mean blood pressure to heart rate and blood pressure to renal sympathetic nerve activity. In conclusion, ketamine and propofol had different effects on autonomic cardiovascular function, but attenuated the baroreflex sensitivity of heart rate and renal sympathetic nerve activity in a dose-dependent manner. These results suggest the possibility that baroreflex sensitivity may reflect the depth of anethesia. PMID- 11476281 TI - Slow inactivation of sodium currents in the rat nodose neurons. AB - Nodose neurons express sodium currents that can be differentiated based on their sensitivity to tetrodotoxin. Several studies have demonstrated significant differences in voltage-dependence and kinetics of activation and inactivation between tetrodotoxin-sensitive and tetrodotoxin-resistant currents. However, little is known about the slow inactivation. Using whole cell patch-clamp technique fast and slow inactivation of sodium currents were studied in cultured rat nodose neurons. Tetrodotoxin-resistant currents recovered much more rapidly after a 15-ms depolarization than tetrodotoxin-sensitive currents. However, repeated 5-ms depolarizations at 10 Hz induced a cumulative inhibition that was more prolonged in tetrodotoxin-resistant compared to tetrodotoxin-sensitive currents. Consistent with these findings, slow inactivation proceeded more rapidly and was more complete for the tetrodotoxin-resistant than for tetrodotoxin-sensitive currents. While the voltage-dependence of fast inactivation differed significantly between the pharmacologically distinct currents, the voltage-dependence of slow inactivation was similar for both sodium currents. We conclude that slow inactivation of sodium currents can be triggered by trains of brief depolarizations. The resulting prolonged decrease in membrane excitability may contribute to the different patterns of action potential generation observed in primary afferent neurons. PMID- 11476282 TI - Pancreatic exocrine responses to parasympathetic stimulation in anaesthetized pigs. AB - Pancreatic exocrine responses to stimulation of the peripheral ends of the vagus nerves intermittently have been investigated in anaesthetized pigs and compared with the effects of continuous stimulation at corresponding frequencies. At relatively low frequencies < or =20 Hz in bursts or 2 Hz continuously) both the flow of pancreatic juice and the output of protein therein were potentiated by stimulating in bursts. Thus stimulation at 20 Hz in bursts produced a significantly greater flow of pancreatic juice than stimulation at 2 Hz continuously (10.9+/-0.9 compared to 4.8+/-0.7 microl min(-1) (g gland)-1 , respectively; P<0.01). Likewise the output of protein during intermittent stimulation at 20 Hz (144+/-23 microg min(-1) (g gland)-1) far exceeded that produced during continuous stimulation at 2 Hz (49+/-9 microg min(-1) (g gland) 1; P<0.01). Both differences were abolished by atropine (0.5 mg kg(-1) i.v.), which augmented the flow during continuous stimulation (to 8.7 +/- 1.5 microl min(-1) (g gland)-1; P<0.05 at 2 Hz) and substantially reduced the output of protein during intermittent stimulation (to 27+/-7 ng min(-1) (g gland)-1; P<0.01 at 20 Hz in bursts). These results show that a variety of pancreatic exocrine responses can be enhanced by stimulating the parasympathetic innervation in bursts. They are also consistent with the contention that the secretion of protein from the gland, in response to parasympathetic stimulation, is dependent mainly on activation of muscarinic receptors. They confirm that the flow of pancreatic juice is due mainly to the release of VIP and show that, in the absence of atropine, this is restricted by muscarinic inhibition which may be presynaptic as elsewhere. PMID- 11476283 TI - Increased levels of hypothalamic neuronal nitric oxide synthase and vasopressin in salt-loaded Dahl rat. AB - The plasma concentration of arginine vasopression (AVP) and the expression level of the neuronal nitric oxide synthase (nNOS) gene in the paraventricular nucleus (PVN) and the Supraoptic nucleus (SON) of Sprague-Dawley (SD). Dahl salt sensitive (S) and Dahl salt-resistant (R) rats on a high salt diet were examined by radioimmunoassay for AVP and in situ hybridization histochemistry for nNOS. The high salt diet containing 8.0% NaCl was given for 4 weeks. The concentrations of AVP in hypertensive Dahl S rats were significantly increased in comparison with those in SD rats and Dahl R rats on a high salt diet. The levels of nNOS mRNA and NADPH-diaphorase activity in the PVN and SON of hypertensive Dahl S rats were greater than those in Dahl R rats on a high salt diet. The antihypertensive drugs, either nicardipine or captopril were administered to the Dahl S rats for 2 weeks beginning 2 weeks after the start of the high salt diet The nNOS mRNA in the PVN and SON of Dahl S rats given a high salt diet was not upregulated by treatment with nicardipine, while the nNOS mRNA in salt loaded Dahl S rats was greater upregulated by treatment with captopril to that greater than without the antihypertensive drug. Our results suggest that the increased NO production in the PVN and SON of hypertensive Dahl S rats may be ineffective in decreasing blood pressure or inhibiting AVP secretion. PMID- 11476284 TI - Neurons in amygdala mediate ear pinna vasoconstriction elicited by unconditioned salient stimuli in conscious rabbits. AB - We determined whether functional integrity of neurons in the amygdala is necessary for sudden episodes of cutaneous vasoconstriction that occur when the conscious animal detects a salient alerting stimulus. To inhibit neuronal function, muscimol (5 nmol in 300 nl), a long acting and potent GABA-A receptor agonist that hyperpolarizes neurons, was injected bilaterally into the amygdala or into a more dorsal control site in conscious rabbits. Cutaneous blood flow was measured in the ear pinna flow using an ultrasonic Doppler probe chronically implanted around the central ear artery. Ear flow responses to salient unconditioned alerting stimuli (fur touch, slight cage movement. removal of drape covering cage) were examined before and after injection of the muscimol, and the effects compared with effects of muscimol on the ear flow response to more nociceptive stimuli, including ear pinch. Muscimol injections into the dorsal control site did not significantly alter alerting-related episodes of ear pinna vasoconstriction. Muscimol injections into the amygdala almost completely abolished ear vasoconstriction elicited by fur touch (0/5 positive responses), drape removal (0/7 positive responses) and cage movement (0/7 positive responses). Muscimol injections into the amygdala reduced the mean ear flow coefficient of variation for a 15 min observation period from 47+/-5 before injection to 15+/-33% after injection (P<0.01, n=7 rabbits). Muscimol injections into the amygdala did not alter the vigorous ear pinna vasoconstriction elicited by ear pinch (7/7 positive responses). Our results indicate that neuronal function in the amygdala, probably the central nucleus of the amygdala, is necessary for the occurrence of ear pinna vasoconstriction episodes elicited by unconditioned salient stimuli but not for the occurrence of corresponding vasoconstriction elicited by nociceptive stimuli. PMID- 11476285 TI - Effect of cervical vagotomy on sympathetic nerve responses to peripheral interleukin-1beta. AB - Although the vagus nerve is an important neural pathway mediating immune-to-brain communication, the role of the vagus in mediating sympathetic nerve discharge (SND) responses to peripheral cytokines is not well established. In the present study we determined renal, interscapular brown adipose tissue (IBAT), splenic, and lumbar SND responses before and for 60 min after the intravenous administration of interleukin-1beta (IL-1beta, 100 ng) in chloralose anesthetized, sham-vagotomized and cervical-vagotomized (bilateral) rats. In sham vagotomized rats, IL-1beta administration increased (P<0.05) splenic and lumbar SND while renal and IBAT SND remained unchanged from control levels. Renal, splenic, and lumbar SND were increased (P<0.05) whereas IBAT SND remained unchanged from control after IL-1beta in vagotomized rats. Renal, splenic, and lumbar SND responses were significantly higher after IL-1beta in vagotomized compared with sham-vagotomized rats. These results demonstrate that regionally selective SND (renal, splenic, and lumbar) responses to IL-1beta can occur in the absence of the vagus nerve and suggest that the vagus nerve provides a tonic inhibition to the discharges in these nerves in response to peripheral IL-1beta. PMID- 11476286 TI - Mechanical responses evoked by nerve stimulation in gastric muscles of mouse lacking inositol trisphosphate receptor. AB - Alteration of mechanical responses elicited by transmural nerve stimulation (TNS) was investigated in pylorus muscle of stomach isolated from mutant mice lacking expression of IP, type-1 receptor. In wild and mutant mice. TNS inhibited spontaneous contractions and generated an off-response at the cessation. The effects of inhibitors of neurotransmission revealed that in wild mice, acetylcholine and nitric oxide were involved as excitatory and inhibitory mediators, respectively. In mutant mice, a lack of nitroxidergic component with associated attenuation of cholinergic transmission was found. The off-response was inhibited by apamin in both mice. In mutant mice, spantide-sensitive excitatory response appeared in the presence of apamin. Acetylcholine and substance P enhanced while noradrenaline and sodium nitroprusside inhibited spontaneous contractions, in both wild and mutant mice; the actions were weaker in mutant mice than in wild mice for any agonists. The results indicate that pylorus smooth muscles receive cholinergic excitatory and nitroxidergic and non adrenergic non-cholinergic inhibitory projections, and a lack of IP, type-1 receptor results in an impairment of cholinergic and nitroxidergic components, with no alteration of non-adrenergic non-cholinergic inhibitory projections. In addition, the mutation induces a substance P projection which is not detected in wild mice. PMID- 11476287 TI - Smoothed pseudo Wigner-Ville distribution as an alternative to Fourier transform in rats. AB - Techniques for examining signals in the time and frequency domains are well established tools. These tools have their limitations; they tell us in a broad sense where the signal component exists in the frequency domain, but they do not tell us how its frequency characteristics change over time. The time-frequency has become a powerful alternative for the analysis of signals. Among various time frequency distribution methods, one of the most studied is the Wigner-Ville distribution. The aim of this study was to evaluate in conscious rats smoothed pseudo Wigner--Ville distribution (SPWVD) as an alternative to the fast Fourier transform (FFT) in RR intervals and in systolic blood pressure (SBP), before and after adrenergic and cholinergic receptor blockade. Fourteen Wistar rats equipped with telemetry probe were evaluated: (1) under control conditions; (2) after injection of saline (100 microl kg(-1) i.v.); (3) after atenolol (1 mg kg(-1) i.v.); (4) after atropine methyl nitrate (0.5 mg kg(-1) i.v.); and (5) after phentolamine (5 mg kg(-1) i.v.). FFT and SPWVD were applied to RR intervals and SBP time series. Six-minute time series of RR intervals, systolic and diastolic pressures were analysed. The bias and distribution of differences between FFT and SPWVD methods in RR intervals under base conditions were 1.4+/-0.4% (r2=0.94; P<0.01) in LF/LF+HF: 1.5+/-0.5% ( r2=0.92; P<0.01) in HF/LF+HF and 4.8+/-1.9% (r2=0.92; P<0.01) in LF/HF. In SBP the bias and distribution were 1.5+/-0.8% (r2=0.90) P<0.05) in LF/LF+HF and 1.7+/-0.6% (r2=-0.92; P<0.01) in HF/LF+HF. In the frequency domain analysis of RR intervals and SBP there was no difference between FFT and SPWVD. The agreement between the methods demonstrates that in stationary signals both methods can be used interchangeably. SPWVD may be an interesting tool to analyse biomedical signals; it provides a good resolution at high frequency and a good frequency resolution at low frequencies independently if signals remain stationary. PMID- 11476288 TI - Increasing homocysteine levels and diabetic autonomic neuropathy. AB - OBJECTIVE: To determine if hyperhomocysteinemia is a risk factor for the development of diabetic sensorimotor peripheral neuropathy (DSPN) and diabetic autonomic neuropathy (DAN). BACKGROUND: Hyperhomocysteinemia and non-insulin dependent diabetes mellitus (NIDDM) are both associated with premature vascular disease. Microvascular ischemia may be a risk factor for DSPN and DAN; therefore, the relationship of hyperhomocysteinemia to DSPN and DAN was investigated. METHODS: Baseline neurological tests and homocysteine levels were determined in patients from a large prospective study of diabetic complications, the Appropriate Blood Pressure Control in Diabetes (ABCD) Trial. RESULTS: Total homocysteine (tHcy) was independently associated with DAN; for each 1 micromol/l increase in tHcy, there was a 7.1% increased risk of developing DAN (P<0.05). There was no association between tHcy and DSPN. CONCLUSIONS: Hyperhomocysteinemia may be a risk factor for DAN but not for DSPN. This relationship may be related to differential small fiber injury. Further studies are needed to investigate this relationship between tHcy and DAN. specifically whether treatment of hyperhomocysteinemia may modify DAN. PMID- 11476289 TI - Effect of skin sympathetic response to local or systemic cold exposure on thermoregulatory functions in humans. AB - We studied how, sympathetic response to cold exposure determines thermoregulatory function. Three female and seven male volunteers (age, 23.2+/-1.9 years) were exposed to abrupt local cooling and gradual systemic cooling with recording of microneurographic skin sympathetic nerve activity tSSNA), skill temperatures (Ts), tympanic temperature (Tty), skin blood flow measured by laser Doppler flowmetry, and sweating rate measured with a ventilated capsule. Local cooling induced an abrupt vasoconstrictor SSNA increase and Tty rise. There was a significant positive correlation between the increase in the vasoconstrictor SSNA and the change rate of Tty. Systemic cooling at 0.2 degrees C/min enhanced SSNA but gradually decreased Tty, and a significant negative correlation was observed between them. A 10-min delay separated the SSNA rise from the subsequent Tty rise following local cooling. A delay of less than 1 min preceded the SSNA increase after the Tty fall induced by systemic cooling. These findings suggested that subjects with a good SSNA response to cold stress can maintain core temperature, but 10 min is necessary to raise the core temperature by reducing heat loss from the skin surface. In contrast. vasoconstrictor SSNA responds linearly to a fall in core temperature with a delay of less than 1 min. PMID- 11476290 TI - Micturitional disturbance in subacute myelo-optico-neuropathy (SMON). AB - BACKGROUND: Micturitional disturbance is known to occur in subacute myelo-optico neuropathy (SMON). However, its pathophysiology is uncertain and few data are available concerning urodynamic findings. We described our results of micturitional histories and urodynamic studies in patients with SMON. METHODS: A history of urinary symptoms was obtained from six patients with SMON (two men, four women; age, 49-72 years, mean 60 years; duration of illness, 14-25 years, mean 19 years). All patients underwent urodynamic studies including measurement of post-micturition residuals, urethral pressure profilometry, cystometry and simultaneous sphincter electromyography. RESULTS: All patients had micturitional symptoms including voiding symptoms in four, filling symptoms in four and urge urinary incontinence in a patient. Urodynamic studies revealed an increased maximum urethral closure pressure in two of four patients studied, decreased bladder volume at first sensation in two, detrusor hyperreflexia in three, absent bulbocavernosus reflex in a patient, and none had detrusor-sphincter dyssynergia or post-micturition residuals. Repeated urodynamic study (10 years after initial study) in a patient with detrusor hyperreflexia showed the same findings. CONCLUSION: Our results indicate that supranuclear pelvic nerve dysfunction to be mainly responsible for the micturitional disturbance in patients with SMON. PMID- 11476291 TI - Skin sympathetic outflow in Buerger's disease. AB - To clarify if sympathetic outflow is altered in Buerger's disease (thromboangitis obliterans, TAO), we measured skin sympathetic nerve activity (SSNA) in TAO patients, and observed the sweating and vasoconstrictive responses during resting and with activating maneuvers. Multiunit postganglionic sympathetic activity was recorded in a skin fascicle of the tibial nerve innervating the skin of the sole (glabrous skin) and peroneal nerve innervating the skin of the dorsum pedis (hairy skin) from five TAO patients and five healthy subjects simultaneously with skin blood flow and sweat expulsion. TAO patients showed significantly less vasoconstrictor SSNA than healthy subjects (17.0+/-1.9 vs. 31.5+/-5.8 bursts/min, P<0.001). Moreover, we found no relationship between vasoconstrictor SSNA and skin blood flow in some patients, while they were well correlated in healthy subjects. There was no evidence for increased sympathetic activity in TAO patients, and no hypersensitive relationship was found between SSNA and skin blood flow. These observations suggested that these TAO patients exhibiting no relationship between skin blood flow reduction and vasoconstrictor activity might not respond to sympathectomy, which is generally expected to result in an increase in skin blood flow. The absence of increased sympathetic nerve activity provides further indirect evidence of a local vascular abnormality in TAO. PMID- 11476292 TI - Depletion of cholinergic neurons of the medullary arcuate nucleus in multiple system atrophy. AB - The human arcuate nucleus (ArcN) has been considered akin to the pontine precerebellar nuclei. However, there is anatomical, functional, and clinical evidence that the ArcN may be the homologue of chemosensitive areas of the ventral medullary surface involved in ventilatory responses to hypercarbia and cerebrospinal fluid acidosis. Acetylcholine has been involved in mechanisms of central chemosensitivity. Loss of ArcN neurons has been reported in patients with multiple system atrophy (MSA), a disorder characterized by disturbed automatic ventilation, but the neurochemical identity of these neurons is undetermined. We sought to determine whether the ArcN contains cholinergic neurons and whether these neurons are depleted in patients with MSA. Medullae were obtained from six patients with MSA, five patients with Parkinson's disease (PD) and six sex- and age-matched controls. Fifty-micron transverse sections obtained through the mid olivary levels were processed for acetylcholinesterase (AchE), choline acetyltransferase (CAT), and alpha-synuclein immunoreactivity. We found that the ArcN contained CAT-positive neurons. There was a significant decrease in density of cholinergic ArcN neurons in MSA but not in PD patients. alpha-Synuclein containing inclusions were present in the ArcN of MSA patients. Depletion of cholinergic neurons may provide a substrate for disturbances in automatic respiration in MSA patients. PMID- 11476293 TI - Changes in regional vascular resistance in response to microinjection of L glutamate into different antero-posterior coordinates of the RVLM in awake rats. AB - Changes in mean arterial pressure (MAP) and in regional vascular resistance (RVR, hindquarter, mesenteric and renal) induced by microinjection of L-glutamate into three different antero-posterior coordinates of the rostral ventrolateral medulla (RVLM) [1,200-1,600 (microm (n=10), 1,601-2,000 microm (n=12) and 2,001-2,500 microm (n=6) rostral to the obex] were investigated in unanesthetized rats. Guide cannulas directed towards the RVLM were implanted 4 days prior to the experiments. Doppler probes were implanted around the superior mesenteric, inferior abdominal aorta and left renal arteries and a catheter was inserted into the femoral artery and vein 1 day prior to the experiments. Insertion of the injector into the RVLM produced an increase in baseline MAP, which was back to control levels 2 min later, when L-glutamate was microinjected. Microinjection of L-glutamate (1 nmol/30 nl) into the three antero-posterior coordinates of the RVLM produced an increase in MAP associated with a similar increase in hindquarter, mesenteric and renal vascular resistance, which were back to control 1 min later. Saline into the RVLM produced negligible effects on MAP and RVR. These findings suggest that the sympathetic vasomotor neurons involved in the regulation of the regional vascular resistance in rats are not topographically distributed in the antero-posterior coordinates of the RVLM. However, the experimental methods used to evaluate the topographic distribution of sympatho vasomotor neurons in the RVLM and the measurement of the regional blood flow may not be precise enough to detect any possible differences. PMID- 11476294 TI - Graft selection in reconstruction of the anterior cruciate ligament. PMID- 11476295 TI - Revision total elbow replacement using the Souter-Strathclyde prosthesis. AB - The Souter-Strathclyde prosthesis was used in 52 evisions of total elbow replacements (TERs) between August 1986 and May 1997. Of these, 50, carried out in 45 patients, were prospectively followed for a mean of 53 months (14 to 139). The procedure produced reliable relief of pain, and the range of movement was preserved. There was a considerable incidence of adverse events associated with revision (30%), and 12 further procedures have been required. Nonetheless, a revision is the preferred salvage procedure for failed primary arthroplasty in the absence of sepsis. PMID- 11476296 TI - Thermal capsular shrinkage for shoulder instability. Mid-term longitudinal outcome study. AB - We present the short- and medium-term clinical results of thermal shrinkage in selected groups of patients with multidirectional or capsular stretch-type instability. We treated 56 patients (61 shoulders) by laser-assisted capsular shrinkage (LACS) and 34 patients (38 shoulders) by radiofrequency (RF) capsular shrinkage. The two groups were followed for mean periods of 40 months and 23 months, respectively. In the LACS group the mean Walch-Duplay score improved to 90 points 18 months after the operation, but then declined to a plateau of about 80 points; 59% of patients considered their shoulders to be 'much better' or 'better' but there was a failure rate of 36.1%. For the RF group the mean Walch Duplay and Constant scores were 80 points at the various follow-up times; 76.3% of patients considered their shoulder to be 'much better' or 'better'. RF failed in nine shoulders (23.7%). These results match some clinical series of patients with multidirectional instability, undergoing open inferior capsular shift, with a similar rate of failure. We believe that the minimal morbidity involved makes thermal shrinkage a viable alternative to open capsular shift in this difficult group of patients. PMID- 11476297 TI - The radial and posterior interosseous nerves. Results fo 260 repairs. AB - The outcome of 260 repairs of the radial and posterior interosseous nerves, graded by Seddon's modification of the Medical Research Council Special Committee's system, was analysed according to four patterns of injury; open 'tidy', open 'untidy', closed traction, and those associated with injury to the axillary or brachial artery. We studied the effect on the outcome of delay in effecting repair and of the length of the defect in the nerve trunk. Of the 242 repairs of the radial nerve we found that 30% had good results and 28% fair; 42% of the repairs had failed. The violence of injury was the most important factor in determining the outcome. Of the open 'tidy' repairs, 79% achieved a good or fair result, and 36% of cases with arterial injury also reached this level. Most repairs failed when the defect in the nerve trunk exceeded 10 cm. When the repairs were carried out within 14 days of injury, 49% achieved a good result; only 28% of later repairs did so. All repairs undertaken after 12 months failed. Of the 18 repairs of the posterior interosseous nerve, 16 achieved a good result. PMID- 11476298 TI - The application of decision analysis to the surgical treatment of early osteoarthritis of the wrist. AB - Osteoarthritis of the wrist is a complication of a number of common traumatic conditions. Arthrodesis of the radiocarpal joint, proximal row carpectomy and excision of the scaphoid, combined with midcarpal arthrodesis, have all been reported as surgical options. There have been no randomised studies comparing these procedures, and the feasibility of conducting this type of trial is limited. We used decision analysis to compare the three surgical techniques. The variables for the model used were based principally on data from the literature. Extensive sensitivity analyses were carried out to test the impact of the values given to these variables on the outcome of the model. The model indicated that the preferred treatment is proximal row carpectomy. Decision analysis allows a comparison between alternative treatments, when evidence from a randomised trial is lacking or unobtainable. The decision-analysis model may also provide insight into aspects of a problem which would be difficult, or impossible, to evaluate by a cohort study. PMID- 11476299 TI - The carpal tunnel syndrome is a bilateral disorder. AB - We studied, retrospectively, 131 patients who had undergone an open operation for the carpal tunnel syndrome (CTS) in 229 hands. The symptoms were present on both sides in 59% of patients when first seen. Neurophysiological impairment of the median nerve was observed in 66% of the asymptomatic hands, and 73% of patients in this group developed symptoms of CTS after the opposite side had been operated on. Follow-up of patients with unilateral CTS showed that the subsequent development of disease in the unaffected hand is very common. We conclude that CTS is a bilateral disorder and that it becomes more evident as time passes. There is a correlation between the duration of symptoms and bilateral occurrence. PMID- 11476300 TI - Intramuscular haemangioma adjacent to the bone surface with periosteal reaction. Report of three cases and review of the literature. AB - We present three cases of intramuscular haemangioma adjacent to bone in the lower limb. All patients had local pain during the third decade. Plain radiographs showed an irregular or hypertrophic periosteal reaction on the shaft of the fibula and an intramuscular mass adjacent to the bone with inhomogeneous high signal intensity on MRI. These lesions mimic periosteal or parosteal tumours. PMID- 11476301 TI - Ilizarov external fixation for severely comminuted supracondylar and intercondylar fractures of the distal femur. AB - Our aim was to determine the clinical effectiveness and safety of Ilizarov external fixation for the acute treatment of severely comminuted extra-articular and intercondylar fractures of the distal femur. A total of 14 consecutive patients with complex fractures was treated. There were three type-A3, two type C2 and nine type-C3 fractures according to the AO/ASIF system. The mean follow-up was 14 months. Most fractures (13) united primarily at a mean of 16 weeks. One patient with a type-IIIA open fracture had infection and nonunion. The mean range of flexion of the knee at the final follow-up was 105 degrees (35 to 130). We conclude that, in the treatment of comminuted fractures of the distal femur, the Ilizarov fixator is safe and effective in providing stability and allowing early rehabilitation. PMID- 11476302 TI - Skin closure after acute shortening. AB - Z-plasty is used to lengthen scars and wounds. We describe the use of a modified technique to shorten wounds in ten consecutive patients undergoing acute shortening of a limb as part of an Ilizarov procedure. The modified technique gave good exposure, easy closure of the wound and fewer problems with healing than standard incisions. PMID- 11476303 TI - Volumetric analysis of osteonecrosis of the femur. Anatomical correlation using MRI. AB - We tested the accuracy of MRI for the precise quantification of the volume of osteonecrosis in 30 hips (stage III). The values were compared with direct anatomical measurements of the femoral heads obtained after total hip replacement. When the area of osteonecrosis was determined visually, and manually outlined on each slice, the accuracy of the measurement of volume was satisfactory, and the mean absolute deviation between MRI and anatomical measurements was similar to that between two MRI data sets. For ten of the hips which were measured by MRI, both before and after collapse, the volume did not appear to change significantly. Our findings suggest that the volume of osteonecrosis can be determined with accuracy by MRI, both before and after collapse. PMID- 11476304 TI - The effect of preoperative donation of autologous blood on deep-vein thrombosis after total hip arthroplasty. AB - We have assessed the effect of the donation of autologous blood and the preoperative level of haemoglobin on the prevalence of postoperative thromboembolism in 2043 patients who had a total hip arthroplasty. The level of haemoglobin was determined seven to ten days before surgery and all patients had venography of the operated leg on the fifth postoperative day. The number of patients who had donated autologous blood (1037) was similar to that who had not (1006). A significant decrease in the incidence of deep-vein thrombosis (DVT) was noted in those who had donated blood preoperatively (9.0%) compared with those who had not (13.5%) (p = 0.003). For all patients, the lower the preoperative level of haemoglobin the less likely it was that a postoperative DVT would develop. Of those who had donated blood, 0.3% developed a postoperative pulmonary embolism compared with 0.7% in those who had not, but this difference was not statistically significant. No significant difference was found in the requirements for transfusion between the two groups. PMID- 11476305 TI - Is titanium so bad? Medium-term outcome of cemented titanium stems. AB - Early failures have seen titanium fall from favour as a material for cemented femoral stems. Between 1989 and 1994, we performed a prospective review of a consecutive series of 122 cemented total hip replacements using the Ultima straight textured titanium stem, and report the five- to ten-year clinical and radiological outcomes. There were no revisions for loosening of the femoral stem. Revision surgery was undertaken for other reasons such as dislocation, infection and loosening of the cup in 7.3%. Of those patients without revision all but two were satisfied with their hip, with 74% graded good or excellent using a modified Harris hip score. Radiological assessment revealed probable loosening in two. Although slight vertical subsidence was found in one-third of patients it had not progressed to loosening. It is not clear whether this represents debonding. Non progressive radiolucent lines (1 to 2 mm) were present in zone 1 at the cement prosthesis interface in 14.7%. Calcar resorption and hypertrophy around the distal stem were not often seen. With 97% survival at a mean of 7.5 years, the medium-term results of this specific cemented titanium stem are reassuring so far, but we are concerned about debonding and future failure. PMID- 11476306 TI - Total hip arthroplasty using porous-coated femoral components in patients with rheumatoid arthritis. AB - We studied the results of total hip arthroplasty (THA) using AML porous-coated femoral components at a mean follow-up of 11 years in a non-selected, consecutive series of patients with rheumatoid arthritis. We reviewed 64 patients with 82 primary THAs using these components. There were seven men (8 hips) and 57 women (74 hips) with a mean age of 55.1 years (24 to 80) at the time of surgery. Nine patients (11 hips) died before the two-year follow-up. Of the remaining 71 hips, only one stem was revised for aseptic loosening. Survivorship for the stems was 98.1% (95% confidence interval (CI) 94.5 to 100.0) at ten years, using a life table analysis, with revision for any reason as an endpoint. Of the 70 unrevised stems, 66 (94%) had bony ingrowth, while four (6%) were radiologically loose at the most recent follow-up (mean 11.4 years). Our study shows the excellent long term results which can be achieved with porous-coated femoral components in patients with rheumatoid arthritis. PMID- 11476307 TI - Cefuroxime-impregnated cement at primary total knee arthroplasty in diabetes mellitus. A prospective, randomised study. AB - We have performed a prospective single-blinded randomised study to evaluate the role of antibiotic-impregnated cement in the prevention of deep infection at primary total knee arthroplasty (TKA) in patients with diabetes mellitus. We studied prospectively 78 arthroplasties performed for osteoarthritis in such patients. They were randomly separated into two groups. In group 1 (41 knees), cefuroxime-impregnated cement was used while in group 2 (37 knees) cefuroxime was not added to the cement. The preoperative, intraoperative and postoperative management was the same for both groups. The mean follow-up was 50 months (26 to 88). There were no cases of deep infection in group 1, but five (13.5%) occurred in group 2 (p = 0.021). We conclude that cefuroxime-impregnated cement is effective in the prevention of deep infection at primary TKA in patients with diabetes mellitus. PMID- 11476308 TI - The Lubinus patellofemoral arthroplasty. A five- to ten-year prospective study. AB - We present a prospective review of the outcome of 76 Lubinus patellofemoral arthroplasties carried out in 59 patients between 1989 and 1995. At a mean follow up of 7.5 years, 62 knees in the 48 patients were reviewed; 11 patients (14 knees) had died. None was lost to follow-up. The clinical outcome using the Bristol Knee Scoring system was satisfactory in 45% of the cases. Maltracking of the patella, resulting in lateral tilt, subluxation and polyethylene wear, was the most common complication (32%). Revision surgery was carried out in 21 knees (28%) giving a cumulative survival rate of 65% (confidence interval (CI) 49 to 77) at eight years. The survival rate for revision and moderate pain was 48% (CI 36 to 59) at six years. Progression of arthritis was seen in seven cases (9%). In five of these (6.5%), the symptoms were severe enough to need revision surgery. Due to the high proportion of unsatisfactory results, we have discontinued the use of this prosthesis. PMID- 11476309 TI - Timing of the administration of tranexamic acid for maximum reduction in blood loss in arthroplasty of the knee. AB - We studied 99 patients who were undergoing total knee arthroplasty (TKA) to determine the optimum protocol for the administration of tranexamic acid (TNA) in order to reduce blood loss. It decreased by more than 40% after the administration of TNA. The haemostatic effect was greatest when TNA was given preoperatively and on deflation of the tourniquet. There was no increase in the incidence of adverse affects in the patients receiving TNA, compared with a control group. We conclude that two injections of TNA, one given preoperatively and one on deflation of the tourniquet, significantly reduce blood loss without increasing the risk of thromboembolic complications. PMID- 11476310 TI - Manipulation and injection for hallux rigidus. Is it worthwhile? AB - Manipulation of the metatarsophalangeal joint and injection with steroid and local anaesthetic are widely practised in the treatment of hallux rigidus, but there is little information on the outcome. We report the results of this procedure carried out on 37 joints, with a minimum follow-up of one year (mean, 41.2 months). Patients with mild (grade-1) changes gained symptomatic relief for a median of six months and only one-third required surgery. Two-thirds of patients with moderate (grade-2) disease proceeded to open surgery. In advanced (grade-III) hallux rigidus, little symptomatic relief was obtained and all patients required operative treatment. We recommend that joints are graded before treatment and that manipulation under anaesthetic and injection be used only in early (grades I and II) hallux rigidus. PMID- 11476311 TI - Necrotising fasciitis of a limb. AB - Between January 1992 and December 1998, we treated 24 patients with necrotising fasciitis of a limb. There were 15 men and nine women with a mean age of 59.8 years (5 to 86). The infection was usually confused with cellulitis. Exquisite pain and early systemic toxicity were the most consistent clinical features. Diabetes mellitus and hepatic cirrhosis were the most commonly associated medical diseases. One third of the patients died. Those with involvement of the limbs above the knee or elbow on admission had a significantly higher rate of mortality than those with distal lesions (Fisher's exact test, p = 0.027). There was no correlation between mortality and advanced age (Student's t-test, p = 0.22) or between amputation and survival (Fisher's exact test, p = 0.39). PMID- 11476312 TI - Perthes' disease in the adolescent. AB - We studied the natural history of Perthes' disease in 62 children in whom the onset of symptoms was in adolescence. Three patterns of disease were noted, namely, late-onset pattern, segmental collapse, or destructive with failure of revascularisation. In the late-onset pattern, the disease followed the sequence of healing seen in younger children, but adequate epiphyseal remodelling did not occur. Consequently, the femoral head was never spherical after revascularisation. With segmental collapse, early and irreversible collapse of part of the epiphysis occurred with gross deformation of the femoral head. The destructive pattern was characterised by a failure of revascularisation and repair of the avascular epiphysis. The radiological outcome was poor in all three patterns. The poorest clinical results were found in the destructive type which was frequently associated with incapacitating pain requiring arthrodesis or excision arthroplasty within three years of onset of the disease. PMID- 11476313 TI - Posterior rotational intertrochanteric osteotomy of the femur in children and adolescents. Use in residual deformity of the femoral head after treatment for developmental dysplasia of the hip. AB - We describe a method of intertrochanteric osteotomy with posterior rotation of the femoral head and neck. We analysed 45 hips in 44 children and adolescents aged from six to 18 years with residual dysplasia after conservative (35) and operative (10) treatment of developmental dysplasia of the hip complicated by avascular necrosis of the femoral head. In ten, femoral osteotomy was combined with a variety of pelvic procedures. Thirty-seven hips (36 patients) were available for follow-up at a mean of 4 years 5 months (2 to 15 years). Excellent results were obtained in nine, good in 17, fair in seven and poor in four. PMID- 11476314 TI - MRI study of talonavicular alignment in club foot. AB - We studied in vivo the talonavicular alignment of club foot in infants using MRI. We examined 26 patients (36 feet) with congenital club foot. The mean age at examination was 9.0 months (4 to 12). All analyses used MRI of the earliest cartilaginous development of the tarsal bones in the transverse plane, rather than the ossific nucleus. The difference in the mean talar neck angle (44.0 +/- 8.1 degrees) in club foot was statistically significant (p < 0.001) when compared with that of the normal foot (30.8 +/- 5.5 degrees). The difference between the mean angles in the group treated by operation (47.9 +/- 6.7 degrees) and those treated conservatively (40.1 +/- 7.5 degrees) was also statistically significant. The anatomical relationship between the head of the talus and the navicular was divided into two patterns, based on the position of the mid-point of the navicular related to the long axis of the head. In the operative group, 18 feet were classified as having a medial shift of the navicular and none had a lateral shift. In the conservative group, 12 showed a medial shift of the navicular and six a lateral shift. All nine unaffected normal feet in which satisfactory MRI measurements were made showed a lateral shift of the navicular. Club feet had a larger talar neck angle and a more medially deviated navicular when compared with normal feet. This was more marked in the surgical group than in the conservative group. PMID- 11476315 TI - The treatment of recurrent arthrogrypotic club foot in children by the Ilizarov method. A preliminary report. AB - Between 1994 and 1997 we used the Ilizarov apparatus to treat 12 recurrent arthrogrypotic club feet in nine patients with a mean age of 5.3 years (3.2 to 7). After a mean of three weeks (two to seven) for correction of the deformity and 1.5 weeks (one to four) for stabilisation in the apparatus, immobilisation in a cast was carried out for a mean of 14 weeks (7 to 24). The mean follow-up period was 35 months (27 to 57). Before operation there were one grade-II (moderate), eight grade-III (severe) and three grade-IV (very severe) club feet, according to the rating system of Dimeglio et al. After operation, all the club feet except one were grade I (benign) with a painless, plantigrade platform. Radiological assessment and functional evaluation confirmed significant improvement. Two complications occurred in one patient, namely, epiphysiolysis of the distal tibia and recurrence of the foot deformity. These results suggest that our proposed modification of the Ilizarov technique is effective in the management of recurrent arthrogrypotic club foot in young children. PMID- 11476316 TI - Treatment of severe osteochondritis dissecans of the elbow using osteochondral grafts from a rib. AB - We treated a patient with extensive osteochondritis dissecans of the elbow by an osteochondral graft from a rib. It had consolidated seven months after operation. When seen at follow-up, after seven years and eight months, the elbow was free from pain with an improvement in the range of movement of 24 degrees. PMID- 11476317 TI - Gluteal compartment syndrome after total knee arthroplasty with epidural postoperative analgesia. AB - We describe two patients who developed gluteal compartment syndrome after total knee arthroplasty (TKA) carried out under epidural analgesic infusion and light sedation. To our knowledge, this occurrence has not been described previously after TKA. PMID- 11476318 TI - Apoptosis and expression of stress protein (ORP150, HO1) during development of ischaemic osteonecrosis in the rat. AB - Using in situ hybridisation and the terminaleoxynucleotidyl transferase-mediated biotin-dUTP nick end-labelling (TUNEL) reaction in rats with osteonecrosis of the femoral head we have studied the effect of ischaemia on the gene expression of the stress proteins oxygen-regulated protein 150 (ORP150) and haemoxygenase 1 (HO1) and the death mechanism of the cells involved in osteonecrosis. Both ORP150 and HO1 have been reported to have important roles in the successful adaptation to oxygen deprivation. ORP150 and HO1 mRNA expression was induced by ischaemia in osteoblasts and osteocytes. In proliferative chondrocytes, these signals were detected constitutively. During the development of ischaemic osteonecrosis, the mechanism of cell death was apoptosis as indicated by DNA fragmentation and the presence of apoptotic bodies in osteocytes, chondrocytes and bone-marrow cells. After the initial ischaemic event, expression of ORP150 and HO1 mRNA, the TUNEL positive reaction and empty lacunae were found sequentially. These findings were exclusive and may be considered to be markers for each stage in the development of osteonecrosis. PMID- 11476319 TI - Spatial and temporal collagen gene expression in lumbar intertransverse fusion in the rabbit. AB - We have examined the process of fusion of the intertransverse processes and bone graft in the rabbit by in situ hybridisation and evaluated the spatial and temporal expression of genes encoding pro-alpha1 (I) collagen (COL1A1), pro alpha1 (II) collagen (COL2A1) and pro-alphal (X) collagen (COL10A1). Beginning at two weeks after operation, osteogenesis and chondrogenesis occurred around the transverse process and the grafted bone at the central portion of the area of the fusion mass. Osteoblasts and osteocytes at the newly-formed woven bone expressed COL1A1. At the cartilage, most chondrocytes expressed COL2A1 and some hypertrophic chondrocytes COL10A1. In some regions, co-expression of COL1A1 and COL2A1 was observed. At four weeks, such expressions for COLlA1, COL2A1 and COL10A1 became prominent at the area of the fusion mass. From four to six weeks, bone remodelling progressed from the area of the transverse processes towards the central zone. Osteoblasts lining the trabeculae expressed a strong signal for COL1A1. At the central portion of the area of the fusion mass, endochondral ossification progressed and chondrocytes expressed COL2A1 and COL10A1. Our findings show that the fusion process begins with the synthesis of collagens around the transverse processes and around the grafted bone independently. Various spatial and temporal osteogenic and chondrogenic responses, including intramembranous, endochondral and transchondroid bone formation, progress after bone grafting at the intertransverse processes. Bone formation through cartilage may play an important role in posterolateral spinal fusion. PMID- 11476320 TI - Evaluation of accuracy and precision of bone markers for the measurement of migration of hip prostheses. A comparison of conventional measurements. AB - Our aim was to determine whether tantalum m arkers improved the accuracy and/or precision of methods for the measurement of migration in total hip replacement based on conventional measurements without mathematical correction of the data, and with Ein Bild Roentgen Analyse - Femoral Component Analysis (EBRA-FCA) which allows a computerised correction. Three observers independently analysed 13 series of roentgen-stereophotogrammetric-analysis (RSA)-compatible radiographs (88). Data were obtained from conventional measurements, EBRA-FCA and the RSA method and all the results were compared with the RSA data. Radiological evaluation was also used to quantify in how many radiographs the intraosseous position of the bone markers had been simulated. The results showed that tantalum markers improve reliability whereas they do not affect accuracy for conventional measurements and for EBRA-FCA. Because of the danger of third-body wear their implantation should be avoided unless they are an integral part of the method. PMID- 11476321 TI - The posterolateral approach to the distal humerus for open reduction and internal fixation of fractures of the lateral condyle in children. PMID- 11476322 TI - The sulcus angle and malalignment of the extensor mechanism of the knee. PMID- 11476323 TI - Fix and flap: the radical orthopaedic and plastic treatment of severe open fractures of the tibia. PMID- 11476324 TI - Serrated W/M osteotomy. PMID- 11476325 TI - Evidence of ehrlichiosis agents found in ticks (Acari: Ixodidae) collected from migratory birds. AB - Two Ehrlichia pathogens were found in immature Ixodes ricinus (L.) ticks collected from migratory passerine birds in the Curonian Spit area of the Baltic Region of Russia (Kaliningrad enclave). During the spring and fall of 2000, 1,606 passerine birds (eight species) were collected; 6.8% of them (110/1,606) were infested by ticks, and 51.8% (57/110) of tick clusters contained various human pathogenic microorganisms. Human monocytic ehrlichiosis (HME) and human granulocytic ehrlichiosis (HGE) agents were found in 14% (8/57) of cases. Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi sensu stricto were found in 92.9% (53/57) of the ticks. In five out of eight cases, infection of both Ehrlichia and Borrelia were obtained. In one case, a single nymph contained HME, B. afzelii, and B. garinii. Borrelia burgdorferi s.s. and B. afzelii were found together in one pool of four nymphs and one larva. All agents were identified using polymerase chain reaction and species-specific primers. In 8.8% of the ticks collected from birds in the fall and 22% in the spring, pathogens were isolated from attached co-feeding nymphs and larvae. These data demonstrate that Ehrlichia exchange could occur between co-feeding ticks on animals without systemic infection. PMID- 11476326 TI - Detection of dengue viruses in field caught male Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Singapore by type-specific PCR. AB - Field male Aedes aegypti (L.) and Aedes albopictus (Skuse) adults caught from fixed monitoring stations weekly for 1 yr were screened for dengue viruses (DEN 1, DEN-2, DEN-3, and DEN-4). The assay was carried out using a single-step reverse transcription (or transcriptase)-polymerase chain reaction (PCR) (RT-PCR) followed by a semi-nested PCR using an upstream consensus primer and four type specific primers within the nonstructural protein three gene (NS3) of dengue viruses. The diagnostic fragments for DEN-1, DEN-2, DEN-3, and DEN-4 serotypes were of sizes 169, 362, 265, and 426 bp, respectively. Results showed that in Singapore 1.33% and 2.15% of Aedes aegypti and Aedes albopictus adult male mosquitoes, respectively, were positive for dengue viruses. The serotypes detected in male Ae. aegypti was DEN-1 (44%), followed by DEN-2 (22.2%) and DEN-3 (22.2%), and DEN-4 (11.1%). For Aedes albopictus males, the serotype was DEN-4 (38.9%), followed by DEN-2 (33.3%), DEN-3 (16.7%), and DEN-1 (11.1%). PMID- 11476327 TI - Arbovirus surveillance from 1990 to 1995 in the Barkedji area (Ferlo) of Senegal, a possible natural focus of Rift Valley fever virus. AB - Surveillance for mosquito-borne viruses was conducted in Barkedji area from 1990 to 1995, following an outbreak of Rift Valley fever (RVF) virus in southern Mauritania. Mosquitoes, sand flies, and midges were collected from human bait and trapped by solid-state U.S. Army battery-powered CDC miniature light traps baited with dry ice or animals (sheep or chickens) at four ponds. Overall, 237,091 male and female mosquitoes representing 52 species in eight genera, 214,967 Phlebotomine sand flies, and 2,527 Culicoides were collected, identified, and tested for arboviruses in 9,490 pools (7,050 pools of female and 331 of male mosquitoes, 2,059 pools of sand flies and 50 pools of Culicoides). Viruses isolated included one Alphavirus, Babanki (BBK); six Flaviviruses, Bagaza (BAG), Ar D 65239, Wesselsbron (WSL), West Nile (WN), Koutango (KOU), Saboya (SAB); two Bunyavirus, Bunyamwera (BUN) and Ngari (NRI); two Phleboviruses, Rift Valley fever (RVF) and Gabek Forest (GF); one Orbivirus, Ar D 66707 (Sanar); one Rhabdovirus, Chandipura (CHP); and one unclassified virus, Ar D 95537. Based on repeated isolations, high field infection rates and abundance, Culex appeared to be the vectors of BAG, BBK, Ar D 65239 (BAG-like), and WN viruses, Ae. vexans and Ae. ochraceus of RVF virus, Mansonia of WN and BAG viruses, Mimomyia of WN and BAG viruses, and Phlebotomine of SAB, CHP, Ar D 95537, and GF viruses. Our data indicate that RVF virus circulated repeatedly in the Barkedji area. PMID- 11476328 TI - Birds disperse ixodid (Acari: Ixodidae) and Borrelia burgdorferi-infected ticks in Canada. AB - A total of 152 ixodid ticks (Acari: Ixodidae) consisting of nine species was collected from 82 passerine birds (33 species) in 14 locations in Canada from 1996 to 2000. The Lyme disease spirochete Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwaldt & Brenner was cultured from the nymph of a blacklegged tick, Ixodes scapularis Say, that had been removed from a common yellowthroat, Geothlypis trichas L., from Bon Portage Island, Nova Scotia. As a result of bird movement, a nymphal I. scapularis removed from a Swainson's thrush, Catharus ustulatus incanus (Godfrey), at Slave Lake, Alberta, during spring migration becomes the new, most western and northern record of this tick species in Canada. Amblyomma longirostre Koch, Amblyomma sabanerae Stoll, and Ixodes baergi Cooley & Kohls are reported for the first time in Canada. Similarly, Amblyomma americanum L., Arnblyomma maculatum Koch, and ixodes muris Bishopp & Smith are reported for the first time on birds in Canada. After removal of an I. muris gravid female from a song sparrow, Melospiza melodia Wilson, at St. Andrews, New Brunswick, eggs were laid, which developed into larvae, and this new tick-host record demonstrates that birds have the potential to start a new tick population. We conclude that passerine birds disperse several species of ixodid ticks in Canada, and during spring migration translocate ticks from the United States, and Central and South America, some of which are infected with B. burgdorferi. PMID- 11476329 TI - Ability of cellulose acetate and polyacrylamide enzyme electrophoresis to separate 13 species of phlebotomine sand flies (Diptera: Psychodidae) from Venezuela. AB - The detection of cryptic species by biochemical methods indicates that within the phlebotomine fauna morphological data are not always adequate for species diagnosis. Cellulose acetate and polyacrylamide enzyme electrophoresis methods were compared for their effectiveness in identifying 13 species of Venezuelan phlebotomine sand flies and resolving alleles. Eight diagnostic loci unambiguously separated these 13 species of sand flies. Although acrylamide was as effective as cellulose acetate in species separation, differences were detected in the resolution of some alleles. Cellulose acetate identified more alleles at Ak and Fum, and resolved better at Pgm, whereas acrylamide identified more alleles at Gpi, Mdh, and Me. Therefore, erroneous species diagnoses may occur, if diagnostic loci detected by one technique are used by a second technique without adequate reference standards. PMID- 11476330 TI - Experimentally induced heat-shock tolerance in Panstrongylus megistus (Hemiptera: Reduviidae). AB - The survival and molting incidence of fifth-instar nymphs of Panstrongylus megistus (Burmeister, 1835), a vector of Chagas' disease, were investigated following sequential heat shocks in which a mild shock (35 or 40 degrees C, 1 h) preceded a more drastic one (40 degrees C, 12 h). The shocks were separated by 8 , 18-, 24-, or 72-h periods at 28 degrees C. The heat-shock tolerance response was more effective when the first shock was given at 40 degrees C. When the period between shocks was 18 h, the tolerance to sequential shocks (in terms of specimen survival) weakened, which suggested a transient control of the process that enables the organism to circumvent the unfavorable effects of severe shock. In terms of molting incidence, the heat-shock tolerance was only demonstrated when the period between the first shock at 40 degrees C for 1 h and the second shock at 40 degrees C for 12 h was > or = 24 h. These results are the first to demonstrate the induction of heat-shock tolerance in a blood-sucking hemipteran. PMID- 11476331 TI - Characterization of phagocytic hemocytes in Ornithodoros moubata (Acari: Ixodidae). AB - Effects of fetal bovine serum (FBS) and complement on phagocytic activity in Ornithodaros moubata (Murray 1877) hemocytes and protease activity in the hemocytes were examined. At least three morphologically different cell types, granulocytes, plasmatocytes, and prohemocytes, were detected in hemolymph of O. moubata, and granulocytes and plasmatocytes showed phagocytic activity. FBS altered phagocytic activity of granulocytes, and complement affected phagocytic activity of plasmatocytes. Ticks were inoculated with fluorescent polystyrene beads in combination with FBS or complement. The average number of beads in granulocytes was significantly higher in the FBS injected group than the control (P < 0.01). The percentage of bead-ingesting plasmatocytes in complement inoculated ticks was significantly lower than that in heat-inactivated complement inoculated and control ticks (P < 0.05). Proteases of tick hemocytes localized in small granules in the cytoplasm not only in phagocytic hemocytes but also in prohemocytes. Results suggested modulation of tick hemocyte function through serum components, and digestion of phagocytosed foreign bodies in the hemocytes. PMID- 11476332 TI - Human onchocerciasis in the Amazonian area of southern Venezuela: spatial and temporal variations in biting and parity rates of black fly (Diptera: Simuliidae) vectors. AB - We investigated some entomological factors underlying altitudinal prevalence variation in the Venezuelan Amazonia human onchocerciasis focus. Spatial and temporal variation in relative abundance, daily biting rate, proportion of parous flies, and monthly parous biting rate were studied for the three main simuliid vectors (based on their vectorial competence: Simulium oyapockense s.l. Floch & Abonnenc approximately = S. incrustatum Lutz << S. guianense s.l. Wise). Yanomami villages were selected among sentinel communities of the ivermectin control program, representing hypo- to hyperendemicity conditions of infection. Spatial variation was explored via increasing village altitude on two river systems (A: Ocamo-Putaco and B: Orinoco-Orinoquito). Temporal variation was studied between 1995 and 1999 by sampling the biting population during dry and rainy mouths. Environmental variables included monthly rainfall and maximum river height. Simuliid species composition itself varied along the altitudinal and prevalence gradient. S. oyapockense s.l. prevailed below 150 m. Above this altitude and up to 240 m, S. incrustatum and S. guianense s.l. became more frequently and evenly collected along A but not along B, where S. incrustatum remained absent. The daily biting rate of S. oyapockense s.l. was higher during the dry season along A, whereas the converse took place along B. Daily biting rate of S. incrustatum was lowest during early rains. By contrast, the daily biting rate of S. guianense s.l. was highest during this period. There was a significant negative cross correlation between proportion of parous of S. oyapockense s.l. and river height (2 and 3 mo lagged), whereas this variable (1 and 2 mo lagged) was positively correlated with the proportion of parous flies for S. incrustatum. Monthly parous biting rate values suggest that the months contributing most to onchocerciasis transmission in the area are likely to be the dry season and the transition periods between seasons. PMID- 11476333 TI - Effect of permethrin-impregnated nets on exiting behavior, blood feeding success, and time of feeding of malaria mosquitoes (Diptera: Culicidae) in western Kenya. AB - The impact of permethrin-treated bednets on the feeding and house entering/exiting behavior of malaria vectors was assessed in two studies in western Kenya. In one study, matched pairs of houses were allocated randomly to receive bednets or no bednets. Exiting mosquitoes were collected in Colombian curtains hung around half of each house; indoor resting mosquitoes were collected by pyrethrum spray catches. The number of Anopheles gambiae Giles and An. arabiensis Patton estimated to have entered the houses was unaffected by the presence of bednets; Anopheles funestus Giles was less likely to enter a house if bednets were present. Anopheles gambiae and An. funestus were less likely to obtain a blood meal and significantly more likely to exit houses when bednets were present. No difference was detected in An. arabiensis rates of blood feeding and exiting. In a second experiment, hourly night biting collections were done on 13 nights during the rainy season to assess whether village-wide use of permethrin-treated bednets caused a shift in the time of biting of malaria vectors. A statistically significant shift was detected in the biting times of An. gambiae s.l., although the observed differences were small. No change was observed in the hourly distribution of An. funestus biting. Our study demonstrated that, at least in the short-term, bednets reduced human-vector contact and blood feeding success but did not lead to changes in the biting times of the malaria vectors in western Kenya. PMID- 11476334 TI - Analysis of survival of young and old Aedes aegypti (Diptera: Culicidac) from Puerto Rico and Thailand. AB - It generally is assumed that the daily probability of survival of wild adult mosquitoes is independent of age. To test this assumption we conducted mark release-recapture studies in Puerto Rico and Thailand to determine if estimated daily survival rates between two different age cohorts of the dengue vector Aedes aegypti (L.) were the same. Survivorship was estimated with nonlinear regression analysis using bootstrapping to obtain estimates of errors. Initial recapture success of the younger cohort was greater than the older cohort at both locations. Our analysis revealed a significantly greater survival rate for the younger cohort of females in Puerto Rico, and no significant differences between age cohorts in Thailand. For comparison, a traditional approach for analyzing these type of data, linear regression of log-transformed captures over time (exponential model), was used to calculate the probability of daily survival based on slopes of linear regression lines for recaptured mosquitoes. With this method, the estimated daily survival rate of older females (13-23 d old) was significantly greater than survival of younger ones (3-13 d old) in Puerto Rico and Thailand. In addition, short-range movement of mosquitoes was observed in Puerto Rico; maximum dispersal distance detected was 79 m. Survival rates of adult Ae. aegypti may be age-dependent and nonlinear regression analysis is a sensitive approach for comparing patterns of mosquito survival based on mark, single release, multiple recapture data. PMID- 11476335 TI - Temperature effects on the dynamics of Aedes albopictus (Diptera: Culicidae) populations in the laboratory. AB - We investigated how constant temperatures of 22, 24, and 26 degrees C experienced across the full life cycle affected the dynamics of caged populations of Aedes albopictus (Skuse). All cages were equipped with plastic beakers that served as sites for oviposition and larval development. We measured the per capita daily mortality and emergence rates of the adults and size of adult females, and estimated the intrinsic rate of increase (r) and asymptotic density (K) for each caged population. Populations at 26 degrees C had greater intrinsic rates of increase and lower asymptotic densities than populations at 22 and 24 degrees C. Populations at high temperatures initially had greater daily per capita emergence rates, and steeper declines in per capita emergence rate as density increased over the course of the experiment. There was no temperature effect on the size of adult females nor on the per capita daily mortality rate of adults. Results indicated that populations of Ae. albopictus occurring in regions with relatively high summer temperatures are likely to have high rates of population growth with populations of adults peaking early in the season. These populations may attain relatively low peak densities of adults. Populations occurring in regions with low summer temperatures are likely to experience slow, steady production of adults throughout the season with population size peaking later in the season, and may attain higher peak densities of adults. High temperature conditions, associated with climate change, may increase the rate of spread of Ae. albopictus by increasing rates of increase and by enhancing colonization due to rapid population growth. PMID- 11476336 TI - Reproductive physiology of Aedes (Aedimorphus) vexans (Diptera: Culicidae) in relation to flight potential. AB - Total protein, lipid, and glycogen of Aedes vexans (Meigen) were related linearly to body size at eclosion. Starvation after emergence led to the determination of minimal irreducible amounts of protein, lipid, and glycogen and the availability of the teneral reserves, whereas access to sucrose revealed the potential for reserve synthesis. Glycogenesis and lipogenesis increased reserves approximately 10-fold the teneral value within 1 and 2 wk after emergence, respectively. Carbohydrate feeding was an essential behavior before blood feeding and oogenesis commenced. Female flight was tested on a flight mill. Maximal flights of 10-17 km in a single night occurred at 2 wk posteclosion and paralleled maximal reserve syntheses. Comparisons of our laboratory data to host-seeking mosquitoes in the field confirmed our data. The vast majority of maternal lipid was transferred to the yolk when a blood meal was taken, but only a quarter of the blood protein was recovered from mature ovaries. Maternal glycogen was used mainly for flight. Fecundity varied between 20 and 120 eggs per female and was determined largely by body size and blood meal volume. At 27 degrees C maximal egg numbers were produced, but at 22 and 17 degrees C the caloric yolk content was greater. Females from the southern United States were smaller than females from northern areas. However, southern females had similar fecundity as northern females, and their flight performances were similar. Differences in the reproductive physiology between this species and Ae. aegypti were discussed. PMID- 11476337 TI - Aedes albopictus (Diptera: Culicidae): physiological aspects of development and reproduction. AB - Basic ecophysiological data are presented on the development and reproduction of Aedes albopictus (Skuse) that were reared and maintained at four temperatures between 12 and 32 degrees C. Median larval developmental time from hatching to pupation was correlated inversely with temperature, lasting 7 d at 32 degrees C and up to 28 d at 12 degrees C. Duration of the pupal period also varied from 2-3 d at 32 degrees C to 7-12 d at 12 degrees C. This extension of larval development elongated the phagoperiod and gave rise to larger imagoes. Based on wing length measurements, body sizes varied from 10 to 57 mm3 for females and from 10 to 30 mm3 for males. The caloric protein content at emergence showed a linear and significant regression with body size, independent of sex, treatment, or temperature. Teneral lipid content also followed a linear relationship with body size at warmer temperatures, whereas at low temperatures it increased exponentially with body size. Glycogen was always below 10% of the protein or lipid levels. Reserves at emergence determined median adult survival times, which ranged from 16 d at 32 degrees C to 100 d at 17 degrees C. Access to sucrose solution allowed females to increase their teneral glycogen up to fourfold within 1 wk, and their lipids up to 10-fold within 2 wk. Despite abroad variation, the number of mature oocytes (15-110 eggs per female) was correlated positively with body size, but inversely with the rearing and maintenance temperature. Utilization of the blood meal protein for oogenesis ranged between 35 and 50%, again inversely correlated with temperature; absolute compositions per oocyte were 6.3-6.5 mcal of protein but ranged from 5 to 7 mcal of lipid. PMID- 11476338 TI - Induction of oogenesis in mosquitoes (Diptera: Culicidae) by infusion of the hemocoel with amino acids. AB - As done previously with adult females of Culex pipiens pallens Coquillett, a mixture of 17 amino acids was infused into the hemocoel of females of seven anautogenous and one autogenous mosquito species belonging to three genera. In Culex. p. quinquefasciatus Say, Cx. tritaeniorhynchus Giles, Cx. kyotoensis Yamaguti & LaCasse, Aedes albopictus (Skuse), Armigeres subalbatus (Coquillett), and Cx. p. molestus Forskal, which previously had laid autogenously matured first batch of eggs, ovarian development was stimulated and frequently continued to maturity. In most mosquitoes, the number of mature follicles nearly doubled when the period of infusion was extended from 24 to 48 h. Therefore, the two previously indicated roles of amino acids, one to initiate ovarian development and the other to regulate the number of maturing oocytes, were confirmed in these species. In Cx. halifaxii Theobald and Ae. japonicus (Theobald), however, the frequency of activation and maturation of ovaries was low compared with the other species, indicating that those species may require some factors other than an increase in amino acids for normal ovarian development after a blood meal. PMID- 11476339 TI - Systemic activity of the avermectins against the cat flea (Siphonaptera: Pulicidae). AB - Ivermectin has potent systemic activity against numerous species of nematodes and arthropods, but there are some important species in these two groups, such as the cat flea, Ctenocephalides felis (Bouche), that appear to be refractory to it. In an effort to determine if the lack of systemic activity against C. felis is specific to ivermectin, or if it is a class-wide phenomenon, 20 avermectin derivatives were tested in an artificial membrane flea feeding system at concentrations of 20, 10, and 1 microg/ml. Results showed that ivermectin had LC90 and LC50 values against fleas of 19.1 and 9.9 microg/ml, respectively. Only four of the other 19 compounds evaluated possessed both LC90 and LC50 values more potent than ivermectin and even then the advantage was modest. Among those four compounds was a two-fold increase in potency relative to ivermectin when the LC90 values were considered (range, 9.2-10.3 microg/ml) and a two- to eight-fold increase when the LC50 values were examined (range, 1.23-5.26 microg/ml). Neither the possession nor the number of oleandrosyl sugars on the macrocyclic backbone were relevant for additional flea activity because among these four compounds were two disaccharides, a monosaccharide and an aglycone. Also, bond disposition between C-22 and 23 did not contribute to increase in activity because these molecules comprise members with either single or double bonds. One of these avermectin analogs was scaled-up and tested subcutaneously in a dog at >100 times the commercial ivermectin dosage and zero efficacy was observed against the flea. We conclude that even the best in vitro avermectin does not have the in vivo potential to become a commercial oral or subcutaneous flea treatment for companion animals. PMID- 11476340 TI - Entomological investigations of an outbreak of Japanese encephalitis virus in the Torres Strait, Australia, in 1998. AB - Japanese encephalitis (JE) virus first appeared in Australia in 1995, when three clinical cases (two fatal) were diagnosed in residents on Badu Island in the Torres Strait, northern Queensland. More recently, two confirmed human JE cases were reported in the Torres Strait Islands and Cape York Peninsula, in northern Queensland in 1998. Shortly after JE virus activity was detected in humans and sentinel pigs on Badu Island in 1998, adult mosquitoes were collected using CO2 and octenol-baited CDC light traps; 43 isolates of JE virus were recovered. Although Culex sitiens group mosquitoes yielded the majority of JE isolates (42), one isolate was also obtained from Ochlerotatus vigilax (Skuse). Four isolates of Ross River virus and nine isolates of Sindbis (SIN) virus were also recovered from members of the Culex sitiens group collected on Badu Island in 1998. In addition, 3,240 mosquitoes were speciated and pooled after being anesthetized with triethylamine (TEA). There was no significant difference in the minimum infection rate of mosquitoes anesthetized with TEA compared with those sorted on refrigerated tables (2.8 and 1.6 per 1,000 mosquitoes, respectively). Nucleotide analysis of the premembrane region and an overlapping region of the fifth nonstructural protein and 3' untranslated regions of representative 1998 Badu Island isolates of JE virus reveled they were identical to each other. Between 99.1% and 100% identity was observed between 1995 and 1998 isolates of JE from Badu Island, as well as isolates of JE from mosquitoes collected in Papua New Guinea (PNG) in 1997 and 1998. This suggests that the New Guinea mainland is the likely source of incursions of JE virus in Australia. PMID- 11476341 TI - Effects of pyriproxyfen on off-host water-balance and survival of adult lone star ticks (Acari: Ixodidae). AB - Newly engorged nymphs of the lone star tick, Amblyoma americanum (L.), were continuously exposed to 4 microg/cm2 of pyriproxyfen residues in glass vials. Treatment of engorged nymphs (n = 285) resulted in significant molting inhibition, with more than one-fourth (26.7%, n = 76) of nymphs dying before or during ecdysis. Treatment effects were evident among ticks that molted to the adult stage, with 26.7% (n = 76) of females, and 17.9% (n = 51) of males exhibiting moribund physical characteristics (i.e., lethargy; dull, discolored and desiccated cuticles; lacking full locomotor competency). A few molted adult ticks (10 males, four females) were dead upon inspection. Only 11.2% of pyriproxyfen treated, emergent females (n = 32), and 11.5% of treated emergent males (n = 25) from 285 ticks treated as engorged nymphs, exhibited normal physical appearance and possessed a full range of locomotor activity. Treated adult ticks maintained within a desiccating environmental chamber at 0% RH and 23 degrees C, had significantly accelerated whole-body water loss rates in comparison to untreated males and females maintained under the same environmental conditions. Additionally, treated adult ticks maintained under optimal environmental conditions (23 degrees C and >95% RH) sustained 100% mortality within 32 d following assignment to these conditions (or 79 d posttreatment as engorged nymphs), whereas untreated ticks had 0% mortality for the same duration of time. Results demonstrate that continuous exposure of nymphs to pyriproxyfen disrupted molting, and accelerated both whole-body water loss and subsequent mortality among emergent adult ticks. PMID- 11476342 TI - Different activities and footwear influence exposure to host-seeking nymphs of Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae). AB - The relative potential for a person accidentally acquiring host-seeking nymphs of the blacklegged tick, Ixodes scapularis Say, and lone star tick, Amblyomma americanum (L.), while wearing either of two types of footwear, walking, crawling on hands and knees, and sitting on large fallen logs in deciduous woods, was evaluated. Although flag samples indicated substantial populations of I. scapularis nymphs and low to moderate numbers of A. americanum at the study sites, relatively few I. scapularis and fewer still A. americanum nymphs were acquired during 30-s and 5-min walks. Significantly fewer I. scapularis were picked up when boots were, worn with ankles taped (an anti-tick precaution) than when sneakers were worn with socks exposed during 5-min walks, but when thus attired, there was no significant difference between the number of nymphs acquired during 30-s walks. Nymphs of I. scapularis did not appear to accumulate incrementally on footwear or clothing during walks when boots were worn and ankles taped. Crawling for 30 s (approximately 3 m distance) yielded significantly more I. scapularis nymphs than walking for 30 s. During crawling, I. scapularis nymphs were picked up on 58% of the 30-s samples. Most ticks picked up during crawls were on pant legs. When a flannel flag cloth (0.5 by 0.5 m) was appressed to the upper surface of logs suitable to be sat upon by tired hikers, I. scapularis nymphs were found on 87% of the logs and in 36% of the samples. These data indicate that the potential for contact with host-seeking nymphs of I. scapularis occurring at these densities is greatly elevated by engaging in activities that involve contact with fallen logs and close contact of hands and knees with leaf litter. PMID- 11476343 TI - Spotted fever group Rickettsia in Dermacentor variabilis (Acari: Ixodidae) infesting raccoons (Carnivora: Procyonidae) and opossums (Marsupialia: Didelphimorphidae) in Tennessee. AB - Spotted fever group Rickettsia were present in 13 of 1,171 (1.1%) of adult Dermacentor variabilis Say removed from raccoons (Procyon lotor L.) and Virginia opossums (Didelphis virginiana Kerr) in western Tennessee. Spotted fever group Rickettsia were detected by amplification of a 617-bp segment of the citrate synthase gene by polymerase chain reaction. PMID- 11476344 TI - Dynamics of the antibody response in cattle infested by Hypoderma (Diptera: Oestridae) after a treatment with injectable and pour-on ivermectin. AB - The dynamics of the anti-Hypoderma antibody response after a treatment with injectable and pour-on ivermectin against first instars of Hypoderma sp. were determined in naturally infested cows. Cattle were randomly assigned to three groups: group 1, which served as an untreated control; group 2, treated with pour on ivermectin at a dose of 500 microg/kg of body weight, and group 3, which received 200 microg/kg of body weight of ivermectin by subcutaneous injection. Both ivermectins showed total efficacy against L-1 of Hypoderma sp. In untreated control animals, grubs became detectable in the 2 mo after treatment. Indirect enzyme-linked immunosorbent assay (ELISA) showed a significant increase in group 3 antibody levels 1 mo after treatment, whereas in group 2 the increase was not significant. Following the peak of anitibody activity in group 3, antibody levels declined at a faster rate than in those treated topically. In both treated groups, cattle remained serologically positive for 4 mo after ivermectin treatment. Thus, serological surveillance programs for cattle grubs conducted during this posttreatment period will not be influenced by treatment with ivermectin. The persistence of antibody levels after larval destruction makes the indirect ELISA unsuitable for the prediction of the efficacy of treatment. PMID- 11476345 TI - Effects of habitat structure on the retention of Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) adults during drag sampling surveys. AB - The ability of Ixodes scapularis Say and Amblyomma americanum (L.) to remain on drags, once acquired, was tested in sparse and dense shrub layer vegetation. When placed on bottoms of drags, adults of both species remained attached for distances three to four times greater in sparse vegetation compared with dense vegetation. When attached to the tops of drags, the differences in retention of ticks on drags between vegetation densities were significant only for A. americanum. In dense vegetation, drags should be checked at 10-m intervals, whereas in sparse vegetation this distance can be extended to 20 m without significant loss of acquired ticks. PMID- 11476346 TI - Tick (Acari) infestations of bats in New Mexico. AB - A total of 278 bats belonging to 16 species was examined for ticks from various sites in New Mexico from 1994 to 1998. Seven species of bats were parasitized by ticks: larvae of Ornithodoros kelleyi Cooley & Kohls, Ornithodoros rossi Kohls, Sonenshine & Clifford (Argasidae), or both. Both species of ticks are reported from New Mexico for the first time. Infestation prevalences for parasitized bats ranged from 2 to 25% on different host species for O. kelleyi and from 7 to 25% for O. rossi. The pallid bat, Antrozous pallidus, and the big brown bat. Eptesicus fuscus, were parasitized by both tick species. No distinct host specificity was noted for either tick species. PMID- 11476347 TI - Trade-offs in cancer and reproduction. PMID- 11476348 TI - Testicular function following chemotherapy. AB - Testicular dysfunction is a common long-term sequela of cytotoxic chemotherapy used in the treatment of many malignancies. The degree to which testicular function is affected is dose- and agent-dependent. The impact on germinal epithelial function of standard multi-agent regimens used in the treatment of lymphomas has been widely studied. Procarbazine-containing regimens result in azoospermia in the vast majority of patients, but much lesser degrees of long term gonadotoxicity are apparent with the newer forms of chemotherapy. High-dose chemotherapy used as preparation before bone marrow transplant is also associated with irreversible germinal epithelial failure in the majority of men. Treatment of testicular cancer with cisplatin and carboplatin regimens leads to temporary azoo- and oligozoospermia in most men, with a recovery to normospermia in 80% by 5 years. There is also evidence of mild Leydig cell impairment in a proportion of men treated with cytotoxic agents, although the clinical significance of this is not clear. Several methods of preserving testicular function during potentially sterilizing treatment have been considered. At present, sperm banking remains the only proven method, although hormonal manipulation to enhance recovery of spermatogenesis and cryopreservation of testicular germ cells are possibilities for the future. PMID- 11476349 TI - Sperm banking and assisted reproduction treatment for couples following cancer treatment of the male partner. AB - In recent years, the survival of young males suffering from cancer has been improved. Development of new techniques such as IVF and intracytoplasmic sperm injection enables even low quality spermatozoa to be used successfully. It is possible therefore to preserve fertility potential of cancer patients before embarking on adjuvant chemotherapy and radiotherapy. Recognizing the importance of protecting the fertility potential of these young males, we present our recommendations for sperm cryopreservation based on the 11 year experience of Bourn Hall and the British Joint Council for Clinical Oncology consultation report. This paper discusses the options available for patients who recover from cancer to become fathers. In many cases patients are concerned about possible abnormalities and teratogenic risks to their future children who have been conceived naturally or by fertility treatment. The data available in the literature may reassure the medical community that there is no such increased risk. However, due to the relatively small number of children born after such treatment, a long-term follow-up is required. There is an ongoing debate regarding the justification for the programme due to the small number of patients who make use of their banked spermatozoa. The authors believe in the importance of protecting the fertility potential of cancer patients, enabling them to father their genetic children in the future while fighting their illness. PMID- 11476350 TI - Cryopreservation of testicular tissue in young cancer patients. AB - Cryopreservation of testicular tissue might benefit prepubertal boys who must undergo chemotherapy or radiotherapy. Cryopreservation of testicular tissue and testicular cells for intracytoplasmic sperm injection (ICSI) is feasible and widely applied. Testicular tissue from prepubertal boys can also be frozen, by applying techniques used with other tissues and with testicular tissue from adult men before ICSI. Good results have been obtained when propanediol is used as a cryoprotectant, but glycerol has also been used when freezing testicular tissue. Spermatogonia might also be isolated and cryopreserved as a cell suspension, though practical experience in humans is lacking. Transplantation of the frozen thawed cells back to the testes after cancer treatment might result in restoration of spermatogenesis. Live offspring have been born to mice after transplantation of fresh, but not cryopreserved, testicular cells. Transplantation is technically feasible also in larger species, but to date no offspring have been born. Spermatogenesis in vitro would be an excellent option for boys with haematological malignancies who carry a risk of relapse after transplantation; however, at present the method is feasible only for the late stages of spermatogenesis. PMID- 11476351 TI - Cancer in pregnancy: maternal and fetal implications. AB - Cancer is the second leading cause of death in women during their reproductive years, and complicates approximately 0.1% of all pregnancies. When cancer occurs during gestation it poses immense pressure on the pregnant patient, her relatives and her physicians. As cancer is diagnosed during gestation, it raises conflicts between optimal maternal therapy and fetal well-being. In this review, the available data are analysed regarding the impact of pregnancy on the course of the disease, and the effects of the malignant process and its treatment on both the mother and her fetus. Here, attention is focused on the most common malignancies associated with pregnancy; cervical and breast cancer, malignant melanoma and lymphoma. In addition, attention is focused on the available data regarding the impact of cytotoxic and radiation treatments on the mother and fetus. PMID- 11476352 TI - Genetic and teratogenic effects of cancer treatments on gametes and embryos. AB - Male and female germ cells vary in their sensitivity to the mutagenic effects of chemotherapy and radiotherapy, depending on their stage of maturation and the agent used. Although sperm DNA damage exists following treatment, no increase in genetic defects or congenital malformations was detected among children conceived to parents who have previously undergone chemotherapy or radiotherapy. The use of assisted reproductive technologies and micromanipulation techniques might increase this risk; hence caution should be exercised. In female cancer patients, miscarriage and congenital malformations are not increased following chemotherapy. However, when IVF and embryo cryopreservation is practised between or shortly after treatment, possible genetic risks to the growing oocytes exist, and hence the babies should be screened. During pregnancy, the potential teratogenic effects of chemotherapy influence the choice and timing of therapy. Termination is usually recommended in the first trimester. Second- and third trimester exposure does not usually increase the teratogenic risk and cognitive development, but it may increase the risk of poor obstetric outcome and fetal myelosuppression. During the first two weeks after fertilization of the embryo, radiation is lethal but not teratogenic. High doses of radiation during pregnancy induce anomalies, impaired growth and mental retardation, and there may be an increased risk of childhood leukaemia and other tumours in the offspring. PMID- 11476353 TI - Candidate genes in polycystic ovary syndrome. AB - The candidate gene approach has already paid some dividends in trying to understand the complex genetics of polycystic ovary syndrome (PCOS). In terms of steroidogenic abnormalities, CYP11a-encoding P450 side chain cleavage-appears to be a major susceptibility locus. In relation to the well-described metabolic disturbances in PCOS, the insulin gene variable number tandem repeat (VNTR) appears to be a promising candidate, at least in populations studied in the UK. Finally, genes implicated in ovarian follicular development may have a role in the aetiology of PCOS, as demonstrated by recent identification of the follistatin gene as a potential disease locus. It seems unlikely that PCOS can be explained on the basis of a single gene disorder although, in a given family, one gene may have a predominant effect. An oligogenic model seems the most appropriate basis on which to understand the genetic origins of this very common disorder. The candidate gene approach has been useful to date, but it may prove important in the near future to perform an anonymous genome-wide scan to identify hitherto unheralded susceptibility loci. PMID- 11476354 TI - Affected sib-pair analysis in endometriosis. AB - This paper (i) reviews the current clinical and molecular genetic data which strongly suggest that endometriosis has a genetic basis; (ii) outlines the general principles of affected-sib pair analysis; and (iii) describes the Oxford Endometriosis Gene (OXEGENE) Study which aims, using a positional cloning approach, to identify susceptibility genes involved in the development of the disease. PMID- 11476355 TI - Culture of preimplantation embryos and its long-term effects on gene expression and phenotype. AB - A growing number of medical, scientific and biotechnological procedures rely on culture of mammalian preimplantation embryos. This review presents currently available data on aberrant offspring development that sometimes arises from commonly applied in-vitro procedures in humans, ruminant species and mice. Comparison between mammalian species reveals similarities in the phenotypic abnormalities that are observed at fetal and perinatal stages of development. In particular, aberrant effects on fetal growth have been observed in multiple studies in which serum complemented the preimplantation culture medium. Although it remains to be determined whether there is a common causal mechanism(s) involved, several hypotheses have been put forward to account for the variety of the observed developmental abnormalities. One of these postulates that culture can result in the epigenetic deregulation of developmentally important genes, and that such epigenetic alterations would affect in particular the expression of genes that are subject to genomic imprinting. Imprinted genes play key roles in the control of fetal growth, and altered imprinting can cause growth defects. Some recent in-vitro culture studies on mice and ruminant species now lend support to this hypothesis. PMID- 11476356 TI - Cytoplasmic transfer in assisted reproduction. AB - This report details the use of cytoplasmic transfer in human oocytes. The introduction of a small amount of ooplasm from a donor oocyte or zygote may alter the function of oocytes, with probable deficiencies. Cytoplasmic transfer from fertile donor oocytes or zygotes into compromised oocytes from patients with recurrent implantation failure after assisted reproduction has now led to the birth of nearly 30 healthy babies worldwide. Transfer of small amounts of cytoplasm probably involves mRNAs, proteins and mitochondria, as well as other factors and organelles. Even though the use of cytoplasmic transfer has been employed in several IVF clinics--and pregnancies have resulted--it is not known definitively whether the physiology of the early embryo is affected. This review outlines the experimental cytoplasmic transfer techniques and postulates the future impact in assisted reproduction. PMID- 11476357 TI - Who remembers the true Italian Mediterranean diet? PMID- 11476358 TI - Food habits in a southern Italian town (Nicotera) in 1960 and 1996: still a reference Italian Mediterranean diet? AB - BACKGROUND: A follow-up analysis of cohorts surveyed in the "Seven Countries Study" has provided increasing evidence of an association between diet and morbidity or mortality from coronary heart disease (CHD) and cancer. The effects of the "Mediterranean diet" on mortality is still evident in Italy, where food patterns differ significantly in different geographical areas. OBJECTIVE: To examine differences in food habits in Nicotera, one of the Italian rural areas of the Seven Countries Study, between 1960 and 1996. METHODS: In 1996, 80 subjects, 37 females and 43 males, aged 40-59 years, were examined in Nicotera assessing food intake by means of a semiquantitative questionnaire of food frequency, validated for the Italian population. In 1960, food intake of a sample of Nicotera subjects was assessed by weighed record method for three seasons. RESULTS: Food choices differed markedly between 1960 and 1996. Consumption of animal foods increased, as did that of cakes, pies and cookies and sweet beverages in both male and female groups; an increase of alcoholic beverages was observed only in females. CONCLUSIONS: In 1960, Nicotera inhabitants were following a diet defined as a "reference Italian Mediterranean diet", but by 1996 the Nicotera diet approached that of an average Italian diet, whose characteristics fall short of a true Mediterranean diet. This change in dietary habits may be responsible for an increased risk of CHD and cancer in the general population in the absence of other factors. PMID- 11476359 TI - Italian multicentre study of intensive therapy with insulin lispro in 1184 patients with Type 1 diabetes. AB - Insulin lispro is absorbed more rapidly and has a shorter duration of action than regular human insulin. It improves glycaemic control but large-scale studies are required to identify regimens that optimise efficacy and safety with local dietary habits. This study involved 1184 Italian patients with Type 1 diabetes, randomised to insulin lispro (n=586) or regular human insulin (n=598) as pre-meal bolus for 3 months. Optimisation of basal NPH insulin was carried out in both groups. The number of administrations of NPH insulin was increased when using insulin lispro but, because basal and bolus insulins were mixed before meals, the total number of injections per day was unchanged. Compliance to administration time was significantly (p<0.001) greater with insulin lispro than with regular human insulin. Post-prandial blood glucose levels were lower with insulin lispro after breakfast (p<0.001), lunch (p<0.005) and dinner (p<0.001). The HbA1c level was decreased from baseline by both insulins, but the percent increase in patients with acceptable (<8%) HbA1c was greater with insulin lispro. While frequency of hypoglycaemia was decreased from baseline by both insulins, the proportion of episodes classified as severe was significantly increased from baseline with regular human insulin, but not with insulin lispro. Thus, compared with regular human insulin, improved glycaemic control was achieved with insulin lispro without an increase in severe hypoglyeaemia. PMID- 11476360 TI - Lispro insulin in type 1 diabetic patients on a Mediterranean or normal diet: a randomized, cross-over comparative study with regular insulin. AB - Lispro (LP) and regular human (HR) insulins were compared in Type 1 diabetic (T1DM) patients on either a Mediterranean diet or normal diet. Twelve T1DM patients were recruited and randomized into two groups of 6, groups A and B. They were treated in different sequences (in 3-month intervals for 1 year). Group A: LP insulin and normal diet, LP insulin and Mediterranean diet, regular insulin and Mediterranean diet, regular insulin and normal diet. Group B: regular insulin and normal diet, regular insulin and Mediterranean diet, LP insulin and Mediterranean diet, LP insulin and normal diet. Each patient was treated with rapid acting insulin, either LP insulin or HR insulin, before each main meal and a dose of slow acting insulin at bedtime. Every 15 days the glycemic control, the incidence and frequency of hypoglycemic episodes, and any adverse events were evaluated. Every 3 months, hematology and a chemistry panel, pre- and post prandial glycemic and insulinemic profiles were evaluated in all patients. HbA1c levels significantly decreased in LP patients on normal diet, post-prandial glycemic levels were significantly lower in LP than in HR patients from 30 min onwards, 15-min post-prandial insulin levels higher in LP- than in HR-treated patients, and hypoglycemic episodes were significantly less in LP- than in HR treated patients. LP insulin, irrespective of the type of diet, results in more effective glycemic control, significantly reduces hypoglycemic episodes as opposed to traditional insulin therapy and seems to be more effective with a normal diet than with a Mediterranean diet. PMID- 11476361 TI - Influence of patients' representations and beliefs about diabetes and its treatment on their adherence to therapy. AB - Research in the last 25 years has shown that diabetic patients' adhesion to medical advice is a multi-factor phenomenon. Most recent research has focused on a patient-centred approach, on the representations that the patient has with respect to medicine, health and disease (with particular regard to the perceived self-efficiency in managing the disease, trust in conventional medicine, treatment, drugs, etc) and on the level of agreement between the patient's representations and those of the health care provider. These representations can potentially act as barriers/facilitators on patients' adhesion to treatment; the best adherence can only be obtained if the real needs of the patient are met, matching therapy with his/her representations and expectations, and acknowledging the constraints that everyday life puts on the individual. The indication is therefore to avoid exclusively considering the physical burdens of the disease, while ignoring the personal and social significance of the experience that the patient is having. It will take time for this to become routine in health care, since it requires a complex change from a traditional, bio-medical approach to an integrated bio-psycho-social approach. The aim of this review is to show how those disease representations of diabetes, and the treatment having considerable impact on patients' adhesion, are being considered in recent literature, and how this nonetheless still constitutes a little explored aspect in medical consultation and research. PMID- 11476362 TI - Impact responses of the flexed human knee using a deformable impact interface. AB - Blunt impact trauma to the patellofemoral joint during car accidents, sporting activities, and falls can produce a range of injuries to the knee joint, including gross bone fracture, soft tissue injury, and/or microinjuries to bone and soft tissue. Currently, the only well-established knee injury criterion applies to knee impacts suffered during car accidents. This criterion is based solely on the peak impact load delivered to seated cadavers having a single knee flexion angle. More recent studies, however, suggest that the injury potential, its location, and the characteristics of the damage are also a function of knee flexion angle and the stiffness of the impacting structure. For example, at low flexion angles, fractures of the distal patella are common with a rigid impact interface, while at high flexion angles splitting of the femoral condyles is more evident. Low stiffness impact surfaces have been previously shown to distribute impact loads over the anterior surface of the patella to help mitigate gross and microscopic injuries in the 90 deg flexed knee. The objective of the current study was to determine if a deformable impact interface would just as effectively mitigate gross and microscopic injuries to the knee at various flexion angles. Paired experiments were conducted on contralateral knees of 18 human cadavers at three flexion angles (60, 90, 120 deg). One knee was subjected to a fracture level impact experiment with a rigid impactor, and the opposite knee was impacted with a deformable interface (3.3 MPa crush strength honeycomb material) to the same load. This (deformable) impact interface was effective at mitigating gross bone fractures at approximately 5 kN at all flexion angles, but the frequency of split fracture of the femoral condyles may not have been significantly reduced at 120 deg flexion. On the other hand, this deformable interface was not effective in mitigating microscopic injuries observed for all knee flexion angles. These new data, in concert with the existing literature, suggest the chosen impact interface was not optimal for knee injury protection in that fracture and other minor injuries were still produced. For example, in 18 cadavers a total of 20 gross fractures and 20 subfracture injuries were produced with a rigid interface and 5 gross fractures and 21 subfracture injuries with the deformable interface selected for the current study. Additional studies will be needed to optimize the knee impact interface for protection against gross and microscopic injuries to the knee. PMID- 11476363 TI - A frontal plane model of the lumbar spine subjected to a follower load: implications for the role of muscles. AB - Compression on the lumbar spine is 1000 N for standing and walking and is higher during lifting. Ex vivo experiments show it buckles under a vertical load of 80 100 N. Conversely, the whole lumbar spine can support physiologic compressive loads without large displacements when the load is applied along a follower path that approximates the tangent to the curve of the lumbar spine. This study utilized a two-dimensional beam-column model of the lumbar spine in the frontal plane under gravitational and active muscle loads to address the following question: Can trunk muscle activation cause the path of the internal force resultant to approximate the tangent to the spinal curve and allow the lumbar spine to support compressive loads of physiologic magnitudes? The study identified muscle activation patterns that maintained the lumbar spine model under compressive follower load, resulting in the minimization of internal shear forces and bending moments simultaneously at all lumbar levels. The internal force resultant was compressive, and the lumbar spine model, loaded in compression along the follower load path, supported compressive loads of physiologic magnitudes with minimal change in curvature in the frontal plane. Trunk muscles may coactivate to generate a follower load path and allow the ligamentous lumbar spine to support physiologic compressive loads. PMID- 11476364 TI - Elastic and viscoelastic properties of the human pubic symphysis joint: effects of lateral impact joint loading. AB - The human pelvis is susceptible to severe injury in vehicle side impacts owing to its close proximity to the intruding door and unnatural loading through the greater trochanter. Whereas fractures of the pelvic bones are diagnosed with routine radiographs (x-rays) and computerized tomography (CT scans), non displaced damage to the soft tissues of pubic symphysis joints may go undetected. If present, trauma-induced joint laxity may cause pelvic instability, which has been associated with pelvic pain in non-traumatic cases. In this study, mechanical properties of cadaveric pubic symphysis joints from twelve normal and eight laterally impacted pelves were compared. Axial stiffness and creep responses of these isolated symphyses were measured in tension and compression (perpendicular to the joint). Bending stiffness was determined in four primary directions followed by a tension-to-failure test. Loading rate and direction correlated significantly (p<0.05) with stiffness and tensile strength of the unimpacted joints, more so than donor age or gender. The impacted joints had significantly lower stiffness in tension (p <0.04), compression (p<0.003), and posterior bending (p<0.03), and more creep under a compressive step load (p<0.008) than the unimpacted specimens. Tensile strength was reduced following impact, however, not significantly. We concluded that the symphysis joints from the impacted pelves had greater laxity, which may correlate with post-traumatic pelvic pain in some motor vehicle crash occupants. PMID- 11476365 TI - Stability of reconstructed paralyzed shoulders using a reflected long head biceps technique. AB - A new tendon transfer technique is proposed for the reconstruction of the paralyzed shoulders secondary to Brachial Plexus Injury (BPI). In this tendon transfer, the long head of the biceps tendons is utilized as a bridging tendon graft. It is reflected at the exit of the bicipital groove, passed through the deltoid and directed to the trapezius. The technique is referred to here as the Reflected Long Head Bicepts (RLHB) technique. This study evaluated the effect of this tendon transfer on the anterior, posterior, and inferior stability of the reconstructed should using cadaveric specimens. It was shown that loading of the RLHB contributed significantly to anterior stability of the reconstructed shoulder for 90 deg elevation in the scapula plane. The mean displacement was reduced by 56 percent with RLHB loaded (p<0.01), by 56 percent with the rotator cuff loaded (p <0.005), and by 67 percent with both the RLHB and the rotator cuff loaded (p<0.004). For the post-operation conditions, variation of the directions of RLHB had no significant effect on joint displacement in response to anterior loading. The RLHB tendon also contributed to the posterior and inferior stability for the low and middle elevations in the plane of scapula. Two variations of the RLHB tendon transfer procedures, namely the "Sub-Deltoid" and the "Through Deltoid" techniques, were introduced and studied. These two techniques did not seem to have significantly different effects on the displacement of the humeral head in response to both posterior and inferior loading. The results of this study seemed to support the clinical feasibility of this tendon transfer approach as far as the biomedical stability of the reconstruction is concerned. PMID- 11476366 TI - Numerical-experimental method for the validation of a controlled stiffness femoral prosthesis. AB - The aim of this paper is to describe a new numerical-experimental method to determine the stiffness of a conceptual proximal femoral prototype. The methodology consists of the comparison of the numerical and experimental displacement distributions of the prosthesis loaded as a cantilever beam to validate a design concept: controlled stiffness prosthesis. The manufactured prototype used to test the applicability of the numerical-experimental procedure integrates a stiff metal core bonded to a composite material made of an epoxy resin reinforced with carbon-glass braided pre-forms. The prosthesis with an embedded controlled stiffness concept was obtained by varying the geometry of the core with the composite layer thickness. PMID- 11476367 TI - Design and analysis of robust total joint replacements: finite element model experiments with environmental variables. AB - Computer simulation of orthopaedic devices can be prohibitively time consuming, particularly when assessing multiple design and environmental factors. Chang et al. (1999) address these computational challenges using an efficient statistical predictor to optimize a flexible hip implant, defined by a midstem reduction, subjected to multiple environmental conditions. Here, we extend this methodology by: (1) explicitly considering constraint equations in the optimization formulation, (2) showing that the optimal design for one environmental distribution is robust to alternate distributions, and (3) illustrating a sensitivity analysis technique to determine influential design and environmental factors. A thin midstem diameter with a short stabilizing distal tip minimized the bone remodeling signal while maintaining satisfactory stability. Hip joint force orientation was more influential than the effect of the controllable design variables on bone remodeling and the cancellous bone elastic modulus had the most influence on relative motion, both results indicating the importance of including uncontrollable environmental factors. The optimal search indicated that only 16 to 22 computer simulations were necessary to predict the optimal design, a significant savings over traditional search techniques. PMID- 11476368 TI - An in vitro model of neural trauma: device characterization and calcium response to mechanical stretch. AB - An in vitro model for neural trauma was characterized and validated. The model is based on a novel device that is capable of applying high strain rate, homogeneous, and equibiaxial deformation to neural cells in culture. The deformation waveform is fully arbitrary and controlled via closed-loop feedback. Intracellular calcium ([Ca2+]i) alterations were recorded in real time throughout the imposed strain with an epifluorescent microscopy system. Peak change in [Ca2+]i recovery of [Ca2+]i and percent responding NG108-15 cells were shown to be dependent on strain rate (1(-1) to 10(-1)) and magnitude (0.1 to 0.3 Green's Strain). These measures were also shown to depend significantly on the interaction between strain rate and magnitude. This model for neural trauma is a robust system that can be used to investigate the cellular tolerance and response to traumatic brain injury. PMID- 11476369 TI - Anisotropic and inhomogeneous tensile behavior of the human anulus fibrosus: experimental measurement and material model predictions. AB - The anulus fibrosus (AF) of the intervertebral disc exhibits spatial variations in structure and composition that give rise to both anisotropy and inhomogeneity in its material behaviors in tension. In this study, the tensile moduli and Poisson's ratios were measured in samples of human AF along circumferential, axial, and radial directions at inner and outer sites. There was evidence of significant inhomogeneity in the linear-region circumferential tensile modulus (17.4+/-14.3 MPa versus 5.6+/-4.7 MPa, outer versus inner sites) and the Poisson's ratio v21 (0.67+/-0.22 versus 1.6+/-0.7, outer versus inner), but not in the axial modulus (0.8+/-0.9 MPa) or the Poisson's ratios V12 (1.8+/-1.4) or v13 (0.6+/-0.7). These properties were implemented in a linear an isotropic material model of the AF to determine a complete set of model properties and to predict material behaviors for the AF under idealized kinematic states. These predictions demonstrate that interactions between fiber populations in the multilamellae AF significantly contribute to the material behavior, suggesting that a model for th PMID- 11476370 TI - A simple model of bipedal walking predicts the preferred speed-step length relationship. AB - We used a simple model of passive dynamic walking, with the addition of active powering on level ground, to study the preferred relationship between speed and step length in humans. We tested several hypothetical metabolic costs, with one component proportional to the mechanical work associated with pushing off with the stance leg at toe-off, and another component associated with several possible costs of forcing oscillations of the swing leg. For this second component, a cost based on the amount of force needed to oscillate the leg divided by the time duration of that force predicts the preferred speed-step length relationship much better than other costs, such as the amount of mechanical work done in swinging the leg. The cost of force/time models the need to recruit fast muscle fibers for large forces at short durations. The actual mechanical work performed by muscles on the swing leg appears to be of relatively less importance, although it appears to be minimized by the use of short bursts of muscle activity in near-isometric conditions. The combined minimization of toe-off mechanical work and force divided by time predicts the preferred speed-step length relationship. PMID- 11476371 TI - Hemodynamic factors at the distal end-to-side anastomosis of a bypass graft with different POS:DOS flow ratios. AB - A pulsatile flow in vitro model of the distal end-to-side anastomosis of an arterial bypass graft was used to examine the effects that different flow ratios between the proximal outlet segment (POS) and the distal outlet segment (DOS) have on the flow patterns and the distributions of hemodynamic factors in the anastomosis. Amberlite particles were tracked by flow visualization to determine overall flow patterns and velocity measurements were made with Laser Doppler anemometry (LDA) to obtain detailed hemodynamic factors along the artery floor and the graft hood regions. These factors included wall shear stress (WSS), spatial wall shear stress gradient (WSSG), and oscillatory index (OSI). Statistical analysis was used to compare these hemodynamic factors between cases having different POS:DOS flow ratios (Case 1-0:100, Case 2-25:75, Case 3-50:50). The results showed that changes in POS:DOS flow ratios had a great influence on the flow patterns in the anastomosis. With an increase in proximal outlet flow, the range of location of the stagnation point along the artery floor decreased, while the extent of flow separation along the graft hood increased. The statistical results showed that there were significant differences (p<0.05) for the mean WSS between cases along the graft hood, but no significant differences were detected along the artery floor. There were no significant differences for the spatial WSSG along both the artery floor and the graft hood. However, there were significant differences (p<0.05) in the mean OSI between Cases 1 and 2 and between Cases 1 and 3 both along the artery floor and along the graft hood. Comparing these mechanical factors with histological findings of intimal hyperplasia formation obtained by previous canine studies, the results of the statistical analysis suggest that regions exposed to a combination of low mean WSS and high OSI may be most prone to the formation of intimal hyperplasia. PMID- 11476372 TI - The effect of proximal artery flow on the hemodynamics at the distal anastomosis of a vascular bypass graft: computational study. AB - The formation of distal anastomotic intimal hyperplasia (IH), one common mode of bypass graft failure, has been shown to occur in the areas of disturbed flow particular to this site. The nature of theflow in the segment of artery proximal to the distal anastomosis varies from case to case depending on the clinical situation presented. A partial stenosis of a bypassed arterial segment may allow residual prograde flow through the proximal artery entering the distal anastomosis of the graft. A complete stenosis may allow for zero flow in the proximal artery segment or retrograde flow due to the presence of small collateral vessels upstream. Although a number of investigations on the hemodynamics at the distal anastomosis of an end-to-side bypass graft have been conducted, there has not been a uniform treatment of the proximal artery flow condition. As a result, direct comparison of results from study to study may not be appropriate. The purpose of this work was to perform a three-dimensional computational investigation to study the effect of the proximal artery flow condition (i.e., prograde, zero, and retrograde flow) on the hemodynamics at the distal end-to-side anastomosis. We used the finite volume method to solve the full Navier-Stokes equations for steady flow through an idealized geometry of the distal anastomosis. We calculated the flow field and local wall shear stress (WSS) and WSS gradient (WSSG) everywhere in the domain. We also calculated the severity parameter (SP), a quantification of hemodynamic variation, at the anastomosis. Our model showed a marked difference in both the magnitude and spatial distribution of WSS and WSSG. For example, the maximum WSS magnitude on the floor of the artery proximal to the anastomosis for the prograde and zero flow cases is 1.8 and 3.9 dynes/cm2, respectively, while it is increased to 10.3 dynes/cm2 in the retrograde flow case. Similarly, the maximum value of WSSG magnitude on thefloor of the artery proximal to the anastomosis for the prograde flow case is 4.9 dynes/cm3, while it is increased to 13.6 and 24.2 dynes/cm3, respectively, in the zero and retrograde flow cases. The value of SP is highest for the retrograde flow case (13.7 dynes/cm3) and 8.1 and 12.1 percent lower than this for the prograde (12.6 dynes/cm3) and zero (12.0 dynes/cm3) flow cases, respectively. Our model results suggest that the flow condition in the proximal artery is an important determinant of the hemodynamics at the distal anastomosis of end-to-side vascular bypass grafts. Because hemodynamic forces affect the response of vascular endothelial cells, the flow situation in the proximal artery may affect IH formation and, therefore, long-term graft patency. Since surgeons have some control over the flow condition in the proximal artery, results from this study could help determine which flow condition is clinically optimal. PMID- 11476373 TI - Computational fluid dynamics of a vascular access case for hemodialysis. AB - Vascular accesses (VA) for hemodialysis are usually created by native arteriovenous fistulas (AVF) or synthetic grafts. Maintaining patency of VA continues to be a major problem for patients with end-stage renal disease, since in these vessels thrombosis and intimal hyperplasia often occur. These lesions are frequently associated with disturbed flow that develops near bifurcations or sharp curvatures. We explored the possibility of investigating blood flow dynamics in a patient-specific model of end-to-end native AVF using computational fluid dynamics (CFD). Using digital subtraction angiographies of an AVF, we generated a three-dimensional meshwork for numerical analysis of blood flow. As input condition, a time-dependent blood waveform in the radial artery was derived from centerline velocity obtained during echo-color-Doppler ultrasound examination. The finite element solution was calculated using a fluid-dynamic software package. In the straight, afferent side of the radial artery wall shear stress ranged between 20 and 36 dynes/cm2, while on the inner surface of the bending zone it increased up to 350 dynes/cm2. On the venous side, proximal to the anastomosis, wall shear stress was oscillating between negative and positive values (from -12 dynes/cm2 to 112 dynes/cm2), while distal from the anastomosis, the wall shear stress returned within the physiologic range, ranging from 8 to 22 dynes/cm2. Areas of the vessel wall with very high shear stress gradient were identified on the bending zone of the radial artery and on the venous side, after the arteriovenous shunt. Secondary blood flows were also observed in these regions. CFD gave a detailed description of blood flow field and showed that this approach can be used for patient-specific analysis of blood vessels, to understand better the role of local hemodynamic conditions in the development of vascular lesions. PMID- 11476374 TI - Blunt injuries to the patellofemoral joint resulting from transarticular loading are influenced by impactor energy and mass. AB - Various impact models have been used to study the injury mechanics of blunt trauma to diarthrodial joints. The current study was designed to study the relationship between impactor energy and mass on impact biomechanics and injury modalities for a specific test condition and protocol. A total of 48 isolated canine knees were impacted once with one of three free flight inertial masses (0.7, 1.5, or 4.8 kg) at one of three energy levels (2, 11, 22 J). Joint impact biomechanics (peak load, loading rate, contact area) generally increased with increasing energy. Injuries were typically more frequent and more severe with the larger mass at each energy level. Histological analyses of the patellae revealed cartilage injuries at low energy with deep injuries in underlying bone at higher energies. PMID- 11476375 TI - The use of an antifibrosis agent to improve muscle recovery after laceration. AB - Muscle injuries are challenging problems in traumatology and the most frequent injuries in sports medicine. Muscle injuries are capable of healing, although slowly and occasionally with incomplete functional recovery. We observed that lacerated muscle undergoes a rapid process of regeneration, which is hindered by the development of fibrosis. Biologic approaches to enhance muscle regeneration and prevent fibrosis are being investigated to improve muscle healing after injuries. We observed that growth factors can improve muscle regeneration but cannot prevent muscle fibrosis. We investigated the use of an antifibrosis substance, decorin, as an approach to prevent fibrosis and thereby improve muscle healing after injury in murine muscle. We observed that direct injection of human recombinant decorin can efficiently prevent fibrosis and enhance muscle regeneration in the lacerated muscle. More importantly, decorin can improve the recovery of strength in the injured muscle to a level similar to that observed in normal noninjured muscle. These results suggest that injection of decorin improves both the muscle structure and the function of the lacerated muscle to near complete recovery. This study will contribute significantly to the development of strategies to promote efficient muscle healing and complete functional recovery after muscle injuries. PMID- 11476376 TI - Articular cartilage contact pressure after tibial tuberosity transfer. A cadaveric study. AB - Medial transfer of the tibial tuberosity has been commonly used for treatment of recurrent dislocation of the patella and patellofemoral malalignment. In this study, six fresh human cadaveric knees were used. Static intrajoint loads were recorded using Fuji Prescale pressure-sensitive film for contact pressure and contact area determination in a closed kinetic chain knee testing protocol. Peak pressures, average contact pressures, and contact areas of the patellofemoral and tibiofemoral joints were calculated on native intact knee specimens and after tibial tuberosity transfer. All native intact knee specimens had a normal Q angle. Medialization of the tibial tuberosity significantly increased the patellofemoral contact pressure. Medial displacement of the tibial tuberosity also significantly increased the average contact pressure of the medial tibiofemoral compartment and changed the balance of tibiofemoral joint loading. The results of our study suggest that caution should be used when transferring a patellar tendon in the face of a preexisting normal Q angle as this will result in abnormally high peak pressure within the tibiofemoral joint. Overmedialization of the tibial tuberosity should be avoided in the varus knee, the knee after medial meniscectomy, and the knee with preexisting degenerative arthritis of the medial compartment. PMID- 11476377 TI - Meniscal allograft transplantation. Two- to eight-year results. AB - We evaluated 18 of 23 patients who had undergone cryopreserved meniscal allograft transplantation for compartmental pain after total meniscectomy 2 to 8 years (mean, 5.4) after the operation. The Short Form-36 scores revealed a decrease in pain with a significant improvement in function, although function remained limited. There was no significant decrease in joint space on 45 degrees posteroanterior weightbearing radiographs through the duration of the study. Eight of 22 allograft menisci (36%) tore during the study period, necessitating 6 partial and 2 total meniscectomies. Two patients subsequently underwent reimplantation. Histologic examination of the removed tissue revealed reduced cellularity as compared with normal or torn native menisci. Four specimens also underwent detailed cytokine evaluation and demonstrated reduced cytokine expression compared with controls. While successful in alleviating compartmental pain that may be a late consequence of major meniscectomy, allograft menisci are repopulated with fewer cells than are present in normal or torn native menisci. These cells also demonstrate potentially reduced function, as measured by decreased growth factor production. This decreased biologic activity may be a factor that contributes to the high frequency of retears noted in this and prior studies. PMID- 11476378 TI - Medial and lateral meniscal tear patterns in anterior cruciate ligament-deficient knees. A prospective analysis of 575 tears. AB - We used our database to evaluate the locations of meniscal tears associated with anterior cruciate ligament injuries. Five hundred seventy-five meniscal tears were seen in 476 patients. Each tear was categorized prospectively according to the side (medial/lateral), the radial zone of the tear, and the circumferential zone of the tear. The different tear locations were then compared for the medial and lateral menisci and evaluated for statistical significance. We found a nearly equal number of tears on the medial (305) and lateral (270) sides. A significantly greater number of tears on the medial side as compared with the lateral side were posterior (99.4% versus 87.8%) and peripheral (75.4% versus 44.1%). Peripheral posterior horn tears of the medial meniscus were the most common type of tear (230 of 575, 40%) by a statistically significant amount. PMID- 11476379 TI - Initial fixation strength of bioabsorbable and titanium interference screws in anterior cruciate ligament reconstruction. Biomechanical evaluation by single cycle and cyclic loading. AB - We evaluated the initial bone-patellar tendon-bone graft fixation strength of bioabsorbable as compared with titanium interference screws in anterior cruciate ligament reconstruction using matched pairs of porcine knees. Ten pairs underwent single-cycle failure loading at a rate of 50 mm/min, and 10 pairs underwent cyclic loading at half-hertz frequency. The cyclic loading started with 100 load cycles between 50 and 150 N. We then progressively increased loads in 50-N increments after each set of 100 cycles. After 100 cycles at 850 N, the specimens were loaded to failure at a rate of 50 mm/min. In the single-cycle failure loading test, the mean ultimate failure loads (+/-SD) for the bioabsorbable (837 +/- 260 N) and titanium interference screws (863 +/- 192 N) were not significantly different, nor were the mean yield loads or the stiffness of the fixation. In the cyclic loading test, the yield loads were 605 +/- 142 N and 585 +/- 103 N for the bioabsorbable and titanium interference screws, respectively (no significant difference). Although there was no significant difference in the ultimate failure load, more bone block fractures were found in the grafts fixed with a titanium interference screw. Bioabsorbable interference screw fixation thus seems to provide a reasonable alternative to titanium screws. PMID- 11476380 TI - An examination of injuries in major league soccer. The inaugural season. AB - We analyzed the injury data collected from the 10 teams (237 players) that constituted Major League Soccer during its inaugural season. The overall injury rate was 6.2 per 1000 hours of participation. An injury rate of 2.9 per 1000 hours was noted for practice and 35.3 per 1000 hours was found for games. The difference was statistically significant. With regard to severity, 36% of injuries (143 of 399) were categorized as an incident (no time lost from competition). Of the 256 injuries that caused the player to lose playing time, 59% were classified as minor (<7 days lost), 28% as moderate (7 to 29 days lost), and 13% as major (>30 days lost). Seventy-seven percent of the injuries (197 of 256) were to the lower extremity; most frequently affected were the knee (N = 54) and ankle (N = 46). Neither the player's age nor playing position was found to be associated with prevalence or severity of injury. We believe that this information serves to underscore the notion that soccer, as it is played at the professional level, is a sport with a relatively low incidence of injury. PMID- 11476381 TI - Disability days in major league baseball. AB - We have examined the injury experience in Major League Baseball as reflected by the disabled list, based on data presented by American Specialty Companies in their publications, to examine any changes in injury rates over the past 11 years. It is reasonable to expect that improvements in training and conditioning, diagnostic methods, and surgical treatment over the last 11 years would have reduced injuries and resulted in fewer players on the disabled list. Yet, such does not appear to be the case. There is no evidence that the number of injuries in Major League Baseball has declined over the last decade; on the contrary, it appears that both the number of players and player days on the disabled list have increased. Team membership, injury location, and position do not appear to be related to the increase. Nor does it appear that the increase in injuries is a result of more sensitive diagnostic tests allowing the diagnoses of previously unrecognized injuries. Whatever the reason, it is significant that publicly available data, when viewed over an 11-year period, reveal a gradual and consistent increase in reported injuries--suggesting a problem that deserves attention. PMID- 11476382 TI - Head injuries in snowboarders compared with head injuries in skiers. A prospective analysis of 1076 patients from 1994 to 1999 in Niigata, Japan. AB - We investigated snowboarding-related head injury cases and skiing-related head injury cases during five ski seasons at one resort area. There were 634 snowboarding-related head injuries and 442 skiing-related head injuries. The number of snowboarding head injuries increased rapidly over the study period. More male snowboarders than female snowboarders suffered head injuries. For both snowboarders and skiers, head injuries frequently occurred on the easy and middle slopes. Falls were the most frequent causes of injury in both groups. Jumping was a more frequent cause of injury in the snowboarders (30%) than in the skiers (2.5%). Injury to the occipital region predominated in the snowboarders as compared with the skiers. There were 49 organic lesions in 37 snowboarders and 46 organic lesions in 33 skiers. Subdural hematoma was frequent in the snowboarding head injury group, and fracture was frequent in the skiing head injury group compared with the snowboarding group (not significant). Subdural hematoma was likely to be caused by a fall rather than by a collision, and bone fracture was likely to be caused by a collision rather than by a fall. Four snowboarders and one skier died as a result of their head injuries. Our data suggest that snowboarding head injuries may be prevented by protection of the occipital region and refraining from jumping by beginners. PMID- 11476383 TI - Range of motion after Bankart repair. Vertical compared with horizontal capsulotomy. AB - We studied the range of shoulder motion of patients who underwent vertical as compared with horizontal capsulotomies during open Bankart repair for recurrent anterior dislocations of the shoulder. A vertical capsulotomy was used in 10 shoulders and a horizontal capsulotomy was used in 14 shoulders. Except for the method of capsulotomy, the surgical procedure and postoperative rehabilitation were the same. The range of motion was measured at 1.5, 2, 3, 4, 5, 6, 9, and 12 months after the surgery, and at the final follow-up (average, 49 months for the vertical and 26 months for the horizontal group). No dislocations recurred, and the anterior apprehension test was negative in all of the patients in both groups. External rotation in abduction was greater in the horizontal group than in the vertical group; the differences were significantly greater at 9 months and 12 months after surgery and at the final follow-up. External rotation in adduction, flexion, and internal rotation were not significantly different between the groups. We conclude that Bankart repair through a horizontal capsulotomy preserves a better range of external rotation in abduction than does a vertical approach. PMID- 11476384 TI - Radiographic abnormalities in the thoracolumbar spine of young elite skiers. AB - An increased frequency of radiologic abnormalities in the thoracolumbar spine has been reported among young athletes in various sports, but there are no data concerning ski sports. To evaluate the incidence of these abnormalities in young elite skiers, we compared 120 skiers younger than 17 years old (alpine skiers, ski jumpers, and Nordic cross-country skiers) with a random sample of 39 control subjects of the same age who had no history of high-performance sports participation. Standardized anteroposterior and lateral radiographs of the entire lumbar spine, the lower thoracic spine, and the upper part of the sacrum were obtained from each athlete and each control subject. Radiographs were evaluated by two independent observers for the presence and size of anterior and posterior endplate lesions and Schmorl's nodes. The elite alpine skiers and ski jumpers demonstrated a significantly higher rate of anterior endplate lesions than did the control subjects. This finding might be attributable to excessive loading and repetitive trauma of the immature spine under high velocity, especially in the forward bent posture. PMID- 11476385 TI - Techniques for reducing anterior knee symptoms after anterior cruciate ligament reconstruction using a bone-patellar tendon-bone autograft. AB - Seventy-five patients underwent unilateral anterior cruciate ligament reconstruction with an ipsilateral bone-patellar tendon-bone autograft at our institution. The graft was harvested using a two-transverse-incision technique, and patellar and tibial bony defects were repaired with cored bone grafts collected by reaming the femoral socket and the tibial socket or tunnel. We evaluated the incidence of anterior knee pain, donor site tenderness, and sensory disturbance after use of these procedures. We also analyzed the correlation between anterior knee pain and age, sex, bone plug length, range of motion, postoperative stability, patellar tendon shortening, infrapatellar nerve injury, and the size of the patellar defect. Thirteen patients reported anterior knee pain. Donor site tenderness was detected in 10 patients and was located on the inferior pole of the patella, the tibial tubercle, or both. Sensory disturbance was found over the infrapatellar nerve area in 13 patients. Statistical analysis showed that anterior instability (side-to-side difference of >3 mm) and residual patellar bony defect (depth >2 mm) were risk factors for anterior knee pain. The results of our study suggest that cored cancellous bone grafting for complete restoration of the donor site bony defects and the two-transverse-incision technique to preserve the infrapatellar branch of the saphenous nerve contribute to prevention of anterior knee symptoms. PMID- 11476386 TI - Neglected ruptures of the patellar tendon. A case series of four patients. AB - We describe a series of four patients treated for chronic ruptures of the patellar tendon (more than 6 months old). The proximally retracted patella was brought back to its anatomic position, and the tendon stumps were repaired and protected with multiple strands of strong circlage wire, in a figure-of-8 pattern, from the quadriceps tendon to the tibial tubercle. Postoperatively, immediate mobilization without the use of a brace was initiated. The patients averaged 29 months (range, 8 to 80) from the time of injury until definitive treatment and were observed for an average of 27 months (range, 11 to 40) after treatment. Before treatment, all patients had an extensor lag of at least 20 degrees. At initial follow-up, all patients had full active extension with no extensor lag, and this did not deteriorate postoperatively or after wire removal. At the last follow-up, the average flexion was 0 degrees to 112 degrees. This technique avoids the use of autograft or allograft tissue and does not require lengthening of the quadriceps tendon. PMID- 11476387 TI - A comparison of preoperative imaging techniques for predicting patellar tendon graft length before cruciate ligament reconstruction. AB - Autogenous bone-patellar tendon-bone is commonly used as graft material for cruciate ligament reconstructions. If this type of graft is too long, graft fixation other than an interference screw may be required. If it is too short, selection of another type of graft may be necessary. If the length of the patellar tendon portion of the graft could be accurately predicted, preoperative planning could determine the adequacy of this graft and choice of fixation for the planned procedure. Using lateral radiographs with the knee flexed 30 degrees, standard magnetic resonance imaging, and magnetic resonance imaging with supplemental three-dimensional reconstructions, we measured the length of the patellar tendon in cadavers and then compared these measurements with the actual patellar tendon length measurements. Linear regression analysis resulted in r2 values of 0.80, 0.43, and 0.65 for lateral radiographs, standard magnetic resonance imaging, and magnetic resonance imaging with three-dimensional reconstructions, respectively. We concluded that lateral radiographs at 30 degrees of knee flexion are the most accurate predictors of patellar tendon length. PMID- 11476388 TI - Anatomic and biomechanical study of the lateral collateral and popliteofibular ligaments. AB - The position of the fibula around the proximal tibia varies in different people; it was therefore hypothesized that this variation would affect the orientation of the lateral collateral ligament and the popliteofibular ligament complex. This hypothesis was studied in 10 cadaveric knees by measuring the orientation and length changes in these structures as the knee was flexed. The data were correlated with tibiofibular joint position. The strength of the ligaments was also tested. There were significant correlations between fibular head position and ligament orientation with the knee extended. The lateral collateral ligament slackened significantly with knee flexion, whereas the popliteofibular ligament complex did not. The structures became significantly steeper in the sagittal plane as the knee was flexed. The lateral collateral ligament passed through vertical at 70 degrees of knee flexion and was thus poorly oriented to withstand tibial external rotation. The lateral collateral and popliteofibular ligaments had tensile strengths of 309 and 186 N, respectively. The popliteofibular ligament is dominant when the knee is flexed, because of the slackening of the lateral collateral ligament, and so it should always be reconstructed. The anatomic variation causes some knees to have better ligament orientations to withstand posterolateral tibial displacements and, conversely, other knees may be inherently more difficult to stabilize by reconstruction. PMID- 11476389 TI - Graft incorporation within the tibial bone tunnel after anterior cruciate ligament reconstruction with bone-patellar tendon-bone autograft. AB - We described histologic changes in patellar tendon autografts that occur over time within the tibial tunnel in specimens harvested from patients undergoing revision anterior cruciate ligament reconstruction. Ten patients, averaging 21.2 years of age, were divided into two groups based on the time period between their original and revision surgery: early revision (less than 1 year, four patients) and late revision (more than 1 year, six patients). Among the early revision group, the tendon within the tunnel showed increased cellularity and random collagen bundles. A specimen from the shortest early revision case revealed a normal original bone-tendon junction, whereas others showed an obscured structure. Between the tendon and the tunnel wall, granulation tissue was seen and the bone-tendon junction was still immature. In the late revision group, the tendon appeared similar to normal ligament. The original bone-tendon junction was not seen, and the tendon continued completely to the tunnel wall with Sharpey like fibers. Observations in the early revision group suggest that tendon remodeling and bone-tendon integration continue for at least several months after transplantation. The original bone-tendon junction appears to have shifted to the proximal patellar tendon-tunnel wall junction with time. These findings are in agreement with prior animal studies. PMID- 11476390 TI - Patellofemoral stresses during open and closed kinetic chain exercises. An analysis using computer simulation. AB - Rehabilitation of the symptomatic patellofemoral joint aims to strengthen the quadriceps muscles while limiting stresses on the articular cartilage. Some investigators have advocated closed kinetic chain exercises, such as squats, because open kinetic chain exercises, such as leg extensions, have been suspected of placing supraphysiologic stresses on patellofemoral cartilage. We performed computer simulations on geometric data from five cadaveric knees to compare three types of open kinetic chain leg extension exercises (no external load on the ankle, 25-N ankle load, and 100-N ankle load) with closed kinetic chain knee-bend exercises in the range of 20 degrees to 90 degrees of flexion. The exercises were compared in terms of the quadriceps muscle forces, patellofemoral joint contact forces and stresses, and "benefit indices" (the ratio of the quadriceps muscle force to the contact stress). The study revealed that, throughout the entire flexion range, the open kinetic chain stresses were not supraphysiologic nor significantly higher than the closed kinetic chain exercise stresses. These findings are important for patients who have undergone an operation and may feel too unstable on their feet to do closed chain kinetic chain exercises. Open kinetic chain exercises at low flexion angles are also recommended for patients whose proximal patellar lesions preclude loading the patellofemoral joint in deeper flexion. PMID- 11476391 TI - Superior labral strain during the throwing motion. A cadaveric study. AB - We studied the strain on the superior labrum of 10 fresh-frozen cadaveric shoulders with the arm in simulated positions of a pitching motion. We used linear transducers to measure the strain in both the anterior and posterior superior labrum with the arm in various planes and rotations simulating the motions of pitching: early cocking, late cocking, acceleration, deceleration, and follow-through. Predetermined loads, according to the percent of maximum voluntary contraction of the biceps muscle during each phase of pitching, were calculated and applied to the long head of the biceps tendon using a spring device. Only during the late cocking phase, when the arm was in maximal external rotation, was the increase in strain statistically significant for the anterior and posterior portions and the strain on the posterior portion significantly greater than that on the anterior portion of the labrum. The increased strain in the posterior portion may be due to the anatomic orientation of the long head of the biceps tendon at the superior labrum. The increased strain in the late cocking phase may contribute to the detachment of the labrum with the eccentric contraction of the biceps muscle that occurs with rapid extension of the elbow. PMID- 11476392 TI - The influence of tendon length and fit on the strength of a tendon-bone tunnel complex. A biomechanical and histologic study in the dog. AB - Using a dog model, we examined the influence of tendon length and fit within a bone tunnel on the pull-out strength of a tendon-bone tunnel complex at 6 weeks after fixation. Fourteen adult mongrel dogs (weight, 25 to 30 kg) underwent bilateral hindlimb surgery in which the extensor digitorum longus tendon was transplanted into an extraarticular metaphyseal bone tunnel. Our findings demonstrated that pull-out strength at 6 weeks was enhanced by increasing the length of tendon within the tunnel. The average load to failure with 1 cm of tendon within the tunnel was 153.7 +/- 78.6 N, compared with 265.5 +/- 93.3 N for the specimens with 2 cm of tendon in the tunnel. Tendon fit within the tunnel was also found to be important. The average load to failure when a tendon was placed in a 4.2-mm diameter tunnel was 301 +/- 61 N at 6 weeks. The average load to failure when the tendon was placed within a 6-mm diameter tunnel was 228 +/- 65 N. These differences were statistically different. Histologically, the interface between the tendon and bone appeared to be most mature when there was intimate bone-to-tendon contact. These data suggest that maximizing tendon length within a bone tunnel and minimizing tendon-tunnel diameter mismatch will maximize the strength of a tendon-bone tunnel complex at 6 weeks. PMID- 11476393 TI - Stress fractures of the sacrum. An atypical cause of low back pain in the female athlete. AB - Low back pain is a common finding in an athletically active premenopausal female population. We describe an unusual cause of persistent low back/sacroiliac pain: a fatigue-type sacral stress fracture. Plain radiographs, bone scans, computed tomography, and magnetic resonance imaging studies were obtained in the female athletes to determine the nature of the pathologic abnormality. The most significant risk factor for fatigue-type sacral stress fractures was an increase in impact activity due to a more vigorous exercise program. Potential risk factors such as abnormal menstrual history, dietary deficiencies, and low bone mineral density were examined. The clinical course was protracted, with an average 6.6 months of prolonged low back pain before resolution of symptoms. Sacral fatigue-type stress fractures did not preclude the athletes from returning to their previous level of participation once healing had occurred. PMID- 11476394 TI - Acute exercise-induced compartment syndrome in the anterior leg. A case report. PMID- 11476395 TI - Painful subluxation of the acromioclavicular joint. A case report. PMID- 11476396 TI - Failed autologous chondrocyte implantation. Complete atraumatic graft delamination after two years. PMID- 11476397 TI - Hip and groin injuries in athletes. AB - Although athletic injuries about the hip and groin occur less commonly than injuries in the extremities, they can result in extensive rehabilitation time. Thus, an accurate diagnosis and well-organized treatment plan are critical. Because loads of up to eight times body weight have been demonstrated in the hip joint during jogging, presumably even greater loads can occur during vigorous athletic competition. The available imaging modalities are effective diagnostic tools when selected on the basis of a thorough history and physical examination. Considerable controversy exists as to the cause and optimal treatment of groin pain in athletes, or the so-called "sports hernia." There has also been significant recent attention focused on intraarticular lesions that may be amenable to hip arthroscopy. This article briefly reviews several common hip and groin conditions affecting athletic patients and highlights some newer topics. PMID- 11476398 TI - Quantification of posterior capsule tightness and motion loss in patients with shoulder impingement. PMID- 11476399 TI - Tom Hamilton Lecture 2000: journey with emergency medicine. PMID- 11476400 TI - Evidence, expertise and the emergency physician: doctors should anticipate legal reform in their expert evidence. AB - The court system is particularly dependent on expertise in science and medicine. Medical experts should be aware that their legal evidence has the potential to work against justice as there are partisan selection, financial, and adversarial pressures; and laws of evidence which allow legal evidence to be admissible without definition of its medical-evidence base. Currently the main checks on an expert's legal evidence are the professional integrity of the expert, and cross examination. However, there is good reason to expect reform of the laws of legal evidence to define objective criteria for the admissibility of scientific legal evidence. In anticipation of these reforms, emergency physicians appearing in court should address the medical-evidence base of their legal evidence. PMID- 11476401 TI - Guns and roses: working with the International Committee of the Red Cross in southern Afghanistan. PMID- 11476402 TI - Hyperpyrexia in the emergency department. AB - The differential diagnosis of the hyperpyrexic patient in the emergency department is extensive. It includes sepsis, heat illness including heat stroke, neuroleptic malignant syndrome, malignant hyperthermia, serotonin syndrome and thyroid storm. Each of these possible diagnoses has distinguishing features that may help to differentiate one from another. However, establishing the correct diagnosis is a challenge in the setting of the obtunded emergency patient who gives no history and where there may be limited access to any past medical or drug history. This paper presents such a case and reviews the features of the differential diagnoses and management of the hyperpyrexic patient. PMID- 11476403 TI - Spontaneous pneumomediastinum, pneumothorax and ecstasy abuse. AB - A case report is presented of a patient with bilateral pneumothoraces and spontaneous pneumomediastinum associated with ecstasy use. Pneumothorax and spontaneous pneumomediastinum should be considered in an ecstasy user who complains of chest pain, neck pain or shortness of breath. PMID- 11476404 TI - The 'ODD' thing is Australian. PMID- 11476405 TI - Amateur reptile keepers and antivenom consumption. PMID- 11476406 TI - Advance directive knowledge and research appears lacking in Australia: comment. PMID- 11476407 TI - Exercise-associated hyponatremia: a review. AB - This paper reviews the extensive literature on hyponatremia, a common and potentially serious complication of ultra-distance exercise. Fluid overload is the likely aetiology. Fluid intakes are typically high in athletes who develop hyponatremia, although hyponatremia can occur with relatively modest fluid intakes. The development of fluid overload and hyponatremia in the presence of a modest fluid intake raises the possibility that athletes with this condition may have an impaired renal capacity to excrete a fluid load. The bulk of evidence favours fluid retention in the extracellular space (dilutional hyponatremia) rather than fluid remaining unabsorbed in the intestine. Female gender is an important risk factor for the development of hyponatremia. Management and prevention of exercise-associated hyponatremia are discussed. PMID- 11476408 TI - Venom as a source of useful biologically active molecules. AB - In the specialty area of venomology, emergency physicians traditionally have been most interested in the description of a variety of envenomation syndromes and, subsequent to this, the most appropriate investigative and therapeutic strategies to employ when envenomation is present. Taking an alternative viewpoint, in this paper we have reviewed a selection of interesting areas of biomedical research in which venom components are being investigated for their potential as novel therapeutic agents, pesticides and ion-channel probes. In addition, we describe the molecular imaging tools of X-ray crystallography and nuclear magnetic resonance spectroscopy, key techniques in the development of rationally designed therapeutic agents. PMID- 11476409 TI - Impact of a toxicology service on a metropolitan teaching hospital. AB - OBJECTIVE: To evaluate the impact of a toxicology service on a major metropolitan teaching hospital. METHOD: A descriptive comparative study of all patients presenting with poisoning or suspected poisoning 12 months before and after the commencement of a toxicology service. Data on length of stay in the emergency department, disposition, length of stay of admitted patients and substance(s) involved were examined. RESULTS: A total of 1,316 poisoned patients were studied. There was a statistically significant increase in self-poisonings from 612 to 704 (P = 0.002) and in the number of admissions from 113 to 192 (P < 0.05). There was no significant change in emergency department length of stay. The average length of stay for patients admitted under the care of the toxicology service decreased, especially for complicated patients. CONCLUSION: In the first 12 months of operation the toxicology service treated more patients than the 12-month period prior to commencement, achieving a decrease in average length of stay for those patients admitted to the service. The emergency department length of stay was not altered. We surmise that by decreasing average length of stay for the patients under the care of the toxicology service, the net increase in the general pool of unoccupied beds improves bed access for all emergency department patients. PMID- 11476410 TI - Pulse rate over pressure evaluation (ROPE) is useful in the assessment of compensated haemorrhagic shock. AB - OBJECTIVE: To evaluate a calculation of pulse rate over pulse pressure as a method of predicting decompensation in patients with compensated haemorrhagic shock. METHODS: A retrospective cohort study was undertaken enrolling 184 adult victims of major road trauma who were not shocked on presentation to the emergency department. A pulse rate over pressure evaluation was calculated at presentation and vital signs were observed during the first 2 h of emergency department management. RESULTS: Thirty-one patients became shocked during their first 2 h. A ROPE value of greater than 3.0 had a positive predictive value of 53% and a value less than 3.0 had a negative predictive value of 86% for the development of decompensated shock. CONCLUSIONS: ROPE is an easily applied and useful clinical tool with a ROPE of less than 3.0 being predictive of a patient remaining well and a ROPE of above 3.0 being predictive of the patient developing decompensated shock. PMID- 11476411 TI - NTS versus waiting time: an indicator without definition. AB - OBJECTIVE: The National Triage Scale versus waiting time is a key performance indicator for Australasian emergency departments. However, the point at which the clock starts to measure waiting time has not been defined. The aim of this study was to determine how this indicator is measured in Australia, as well as a number of other issues relating to the application of the National Triage Scale. METHODS: A postal survey was sent to the directors of emergency medicine at 147 emergency departments in Australia. RESULTS: There were 105 responses (71.4%). For measurement of the National Triage Scale versus waiting time indicator, 52 (49.5%) started the clock at the time of patient arrival, 33 (31.4%) at the start of triage, and 11 (10.5%) at the end of triage. Seventy-four emergency departments (70.5%) change the National Triage Scale (NTS) after it has been assigned, and approximately half use set codes for given presentations. Only 28 (26.7%) automatically upgrade children one NTS category. The age definition of a child ranged from 3 years to 18 years. CONCLUSIONS: The measurement of this key performance indicator is not consistent across Australia. Therefore, caution is advised when comparing such data between departments. The Australasian College for Emergency Medicine should produce operational definitions relating to the use of the National Triage Scale in order to provide meaningful comparative data. PMID- 11476412 TI - Wilderness and environmental medicine: another subspecialty for emergency medicine? PMID- 11476413 TI - Randomized controlled trial of midazolam premedication to reduce the subjective adverse effects of adenosine. AB - OBJECTIVE: To determine the safety and efficacy of midazolam premedication to minimize the subjective adverse effects of adenosine. METHODS: Double-blind prospective randomized controlled trial of patients presenting to an urban emergency department. Included were a convenience sample of patients who would have received adenosine by the existing department protocol. Exclusion criteria were pregnancy, benzodiazepine allergy, regular benzodiazepine medication, alcoholism, altered mental state (precluding informed consent), and age less than 18 or greater than 65 years. Subjects received either 1.5 mg of intravenous midazolam or normal saline placebo 5 min prior to the administration of adenosine. Side-effect recall was judged by a questionnaire at 1 h and 24 h postadenosine administration. RESULTS: A total of 34 patients were recruited into the trial, 16 in the placebo group and 18 in the midazolam group. The groups were well matched for demographics, treatment and outcome. There was a significant reduction in the midazolam group for complaint scores of palpitations (P = 0.04) and chest pain (P = 0.02) and a trend to reduction in complaint scores for most other parameters. There were no adverse outcomes in any of the patients studied. CONCLUSIONS: Co-administration of midazolam can safely reduce the recall of the unpleasant adverse effects of adenosine. Its use may be most appropriate in patients who are particularly anxious or have had previous adverse experiences with adenosine. PMID- 11476414 TI - Setting the benchmark for research in the management of acute pain in emergency departments. AB - OBJECTIVE: The aim of this study was to define the degree of change on a visual analogue scale that corresponded to a clinically meaningful reduction in the level of pain reported by patients experiencing acute pain. METHOD: Secondary analysis of data was performed from prospective, descriptive studies of 152 adults and 73 children presenting to emergency departments with acute pain. At presentation and at 20 min intervals to a maximum of three measurements, patients marked the level of their pain on a 100 mm, non-hatched visual analogue scale. At each follow up, they also gave a verbal rating of their pain as 'a lot better', 'a little better', 'much the same', 'a little worse' or 'much worse'. Data from the groups reporting that pain was 'a lot better' and 'about the same' were analysed for mean and median absolute change in visual analogue scale and compared. RESULTS: Of 559 pain comparisons, 60 were rated as 'a lot better' and 257 as 'about the same'. The mean absolute change in visual analogue scale for the group reporting that pain was 'a lot better' was 24 mm (median = 19 mm, 95% confidence interval = 17-31 mm). For the group reporting that pain was 'about the same', the mean visual analogue scale change was 0 mm (median = 0 mm, 95% confidence interval = -1 to 1 mm). CONCLUSION: When evaluating treatments for acute pain, a difference in visual analogue scale pain score of less than about 20 mm is unlikely to be clinically meaningful. Studies should be designed and reported relative to this benchmark rather than the minimum clinically significant difference in visual analogue scale pain score. PMID- 11476415 TI - Abdominal injury and the seat-belt sign. AB - OBJECTIVE: To report on: 1. Prevalence of seat-belt sign in motor vehicle accident victims with abdominal injuries; 2. Prevalence of intestinal injuries in patients with seat-belt sign; and 3. Spectrum of abdominal injuries in a population with high usage of three-point restraints. METHODS: A retrospective chart review was conducted in an adult tertiary-referral hospital from January 1992 to August 1998. Patients were identified from International Classification of Disease-9 codes for abdominal wall and intra-abdominal injuries. RESULTS: The seat-belt sign was present in 60/99. The proportion of intestinal injuries in patients with and without seat-belt sign were 9/60 and 0/39, respectively (P = 0.01). In the 25 patients with intra-abdominal injuries, there were 10 hepatic, 8 splenic, 9 intestinal and 4 retroperitoneal injuries. CONCLUSION: The seat-belt sign is indicative of an increased risk of intestinal injury, which is difficult to detect with no single test providing reliable diagnosis. Other intra-abdominal and retroperitoneal injuries may also occur, which are more readily diagnosed on computed tomography scan or focused abdominal utlrasound when available, but are no more frequent in patients with the seat-belt sign than those without. PMID- 11476416 TI - Non-medical technicians reduce emergency department waiting times. AB - OBJECTIVE: To observe the impact of trained non-medical technicians on emergency department waiting times. The technicians were to perform minor procedures that had previously been performed by medical staff. METHODS: A prospective cohort study with two matched groups of patients. One group comprised patients who presented to the emergency department on days when the technicians worked (working group) and the other comprised patients who presented when the technicians did not work (control group). The waiting times for patients in each group were compared. RESULTS: The median waiting time was 10 min shorter in the working group than the control group (P < 0.0001). This reduction was confined to triage categories 3 and 4. The number of patients who left without being seen was reduced from 8.2% in the control group to 5.3% in the working group (P < 0.00001). CONCLUSION: When added to the normal staff complement, non-medical technicians reduce patient waiting times in the emergency department. PMID- 11476417 TI - Ketamine: do what is right for the patient. PMID- 11476418 TI - Management of the first-time transient ischaemic attack. AB - The assessment and management of patients with a suspected transient ischaemic attack of the brain or eye is a daily task in busy emergency departments. They are common, affecting about 50 per 100,000 population each year. Conditions which mimic a transient ischaemic attack are even more common (e.g. migraine aura, partial seizures, benign paroxysmal positional vertigo, hysteria). This comprehensive review outlines an approach to the management of this complex and challenging problem. PMID- 11476419 TI - Ketamine sedation for children in the emergency department. AB - OBJECTIVE: To introduce a protocol for dissociative sedation using ketamine for children requiring painful procedures within the emergency department. METHOD: A medical literature search was performed along with a review of existing ketamine use protocols in emergency departments from Australia and the United States. Our hospital anaesthetic department and our emergency department nursing staff were closely involved in protocol design. A series of nursing education sessions were held prior to the drug being used within the emergency department. Twenty-eight children aged 1.5-12 years were administered ketamine sedation prior to undergoing a painful procedure. Ketamine was administered either i.m. (dose 3-4 mg/kg) or i.v. (dose 1.00-2.75 mg/kg) depending on physician preference. Midazolam (dose 0.02 mg/kg) and atropine (dose 0.02 mg/kg) were given as adjuncts in the majority of cases. RESULTS: Onset of sedation was rapid (range 1-7 min) for both routes and provided excellent procedural conditions. Discharge time averaged 103 min (range 67-180 min) from time of drug administration. Side effects included vomiting, rash, diplopia, salivation and one case of a distressing emergence reaction. On telephone follow up 1-3 days after the sedation, no delayed adverse events were reported. CONCLUSION: Ketamine is a safe and effective sedative agent for use in children requiring immobilization to enable performance of a painful procedure. It is important to involve both anaesthetic staff and emergency department nursing staff in sedation protocol development to ensure a smooth introduction. We present a protocol for ketamine use in children undergoing painful procedures within the emergency department. PMID- 11476420 TI - Emergency treatment of hypothermia. AB - This review considers several recent concepts regarding aetiology and treatment of accidental hypothermia. The importance and effectiveness of shivering heat production in the attenuation and reversal of hypothermia is described. Immediately following removal from cold stress, the patient is in danger of a deteriorating condition that may be due to collapse of arterial pressure and/or continued decrease of core temperature. Several controversies are discussed. It is advised that, when possible, patients should be actively but gently warmed as soon as possible (especially if arrival at the emergency department will take greater than 45 min). Extra time should be taken to check for life signs before cardiopulmonary resuscitation is initiated. Chest compressions should proceed at regular normothermic rates and care should be taken to not overventilate the patient. In the emergency department, several factors should be considered before deciding on a treatment regimen. These factors include level of consciousness, cardiovascular stability, core temperature and the direction of change of core temperature. It may be advantageous to transport the more severely hypothermic patient to a more advanced care facility even though transport time may be greater. PMID- 11476421 TI - Warfarin toxicity in the emergency department: recommendations for management. AB - OBJECTIVE: To examine patients who presented to a hospital emergency department with evidence of warfarin toxicity, and to review the available published literature to determine what guidelines are available for management of this problem. METHOD: A retrospective analysis of all adult patients who presented to The Geelong Hospital Emergency Department between 1 January 1996 and 30 June 1998 with international normalized ratio > 6 due to warfarin toxicity. RESULTS: A total of 84 patients with international normalized ratio > 6 were included in the study. The average age was 68.3 years. Fifty-three per cent were women. The international normalized ratio was between 6 and 10 in 33 patients (39%), and greater than 10 in 51 patients (61%). Major bleeding occurred in 16.7% of patients, minor bleeding in 17.8%. Sixty-five per cent had no bleeding. Seven patients died, four of those with bleeding. Patients with an international normalized ratio > 10 were more likely to receive fresh frozen plasma (77.6% vs 28.6%; P < 0.001) and in greater amounts (3.0 units vs 0.8 units; P < 0.001) than those with an international normalized ratio of 6-10. There were also more episodes of major bleeding, although not significant. Patients with major bleeding were older (mean 75.4 years vs 67.5 years; P = 0.04), more likely to be admitted (P = 0.046), were more often given fresh frozen plasma (P = 0.003) and in greater amounts (3.28 units vs 2.0 units; P = 0.02). CONCLUSION: Warfarin toxicity is a common problem, and variation in management is not surprising considering the lack of consensus in the literature on this topic. Current recommendations are summarized and a simple flowchart for management of this problem is provided. PMID- 11476422 TI - Implementation and evaluation of an undergraduate emergency medicine curriculum. AB - OBJECTIVE: To describe the implementation and evaluation of an undergraduate course in the first Australian academic emergency medicine unit. METHODS: A descriptive study of a course involving fifth year medical students at the University of Western Australia was undertaken. Teaching included self-directed case problem solving, small group tutorials, practical-skills teaching, clinical attachments and information handouts. Evaluation involved questionnaire scores and written feedback regarding life-support skills, tutorial teaching, course materials, clinical attachments and the course in general. Some groups of students underwent pre-course and post-course examinations. RESULTS: Subjective and objective testing showed that student knowledge significantly improved. Feedback was especially positive toward clinical attachments in emergency departments, practical skills tutorials and the case-based learning method. Students requested longer attachments to emergency departments, and more practical, case-based, interactive and bedside teaching. Problems encountered included inadequate time for teaching, vagueness about student roles and objectives, and dealing with death for the first time without adequate preparation. CONCLUSION: Undergraduate emergency medicine education should become an essential part of Australian and international undergraduate medical education. Emergency medicine is enjoyable and eminently suitable for problem based, interactive and integrated teaching and improves confidence, clinical experience in emergencies, practical skills and teamwork. Improvements include more problem-based teaching, more practical skills sessions and better definition of student roles. These are general principles that can be applied to other undergraduate courses and to designers of other emergency medicine courses, both undergraduate and postgraduate. PMID- 11476423 TI - The treatment of overuse injuries in sports. PMID- 11476424 TI - Eccentric overload training for patients with chronic Achilles tendon pain--a randomised controlled study with reliability testing of the evaluation methods. AB - The purpose was to examine the reliability of measurement techniques and evaluate the effect of a treatment protocol including eccentric overload for patients with chronic pain from the Achilles tendon. Thirty-two patients with proximal achillodynia (44 involved Achilles tendons) participated in tests for reliability measures. No significant differences and strong (r=0.56-0.72) or very strong (r=0.90-0.93) correlations were found between pre-tests, except for the documentation of pain at rest (P<0.008, r=0.45). To evaluate the effect of a 12 week treatment protocol for patients with chronic proximal achillodynia (pain longer than three months) 40 patients (57 involved Achilles tendons) with a mean age of 45 years (range 19-77) were randomised into an experiment group (n=22) and a control group (n=18). Evaluations were performed after six weeks of treatment and after three and six months. The evaluations (including the pre-tests), performed by a physical therapist unaware of the group the patients belonged to, consisted of a questionnaire, a range of motion test, a jumping test, a toe-raise test, a pain on palpation test and pain evaluation during jumping, toe-raises and at rest. A follow-up was also performed after one year. There were no significant differences between groups at any of the evaluations, except that the experiment group jumped significantly lower than the control group at the six-week evaluation. There was, however, an overall better result for the experiment group with significant improvements in plantar flexion, and reduction in pain on palpation, number of patients having pain during walking, having periods when asymptomatic and having swollen Achilles tendon. The controls did not show such changes. Furthermore, at the one-year follow-up there were significantly more patients in the experiment group, compared with the control group, that were satisfied with their present physical activity level, considered themselves fully recovered, and had no pain during or after physical activity. The measurement techniques and the treatment protocol with eccentric overload used in the present study can be recommended for patients with chronic pain from the Achilles tendon. PMID- 11476425 TI - Good clinical results but persisting side-to-side differences in calf muscle strength after surgical treatment of chronic Achilles tendinosis: a 5-year follow up. AB - We have prospectively studied calf muscle strength (isokinetic concentric and eccentric muscle strength at 90 degrees/s and 225 degrees/s of angular velocity in 24 patients (17 males and 7 females, mean age 43.0 years) surgically treated for chronic Achilles tendinosis (at the 2-6 cm level in the tendon). The mean follow-up time was 5 years (range 31-82 months). Surgery was followed by immobilization in a weight-bearing below the knee plaster cast for 2-6 weeks, followed by a stepwise increasing strength training programme. Strength measurements (peak torque) were done preoperatively, and 1 and 5 (mean) years postoperatively. Preoperatively, concentric plantar flexion peak torque at 90 degrees/s and 225 degrees/s and eccentric plantar flexion torque at 90 degrees/s were significantly lower (12.3%, 19.7% and 8.5% respectively) on the injured side compared to the non-injured side. Postoperatively, at the 5-year follow-up, 22 out of 24 patients (92%) were satisfied with the operation and active at their desired level (running, tennis, badminton, walking). There was no significant increase in concentric and eccentric calf muscle strength postoperatively. Concentric plantar flexion peak torque at 90 degrees/s and 225 degrees/s and eccentric plantar flexion peak torque at 90 degrees/s were still significantly lower (7.2%, 8.6% and 8.8% respectively) on the injured side compared to the non injured side. In conclusion it seems that the calf muscle strength deficit seen on the injured side preoperatively in this group of patients remains despite 92% of the patients being pain-free and active in sports or at recreational level after the operation. However, the percentage side-to-side difference is relatively low, and might not have any clinical relevance. PMID- 11476426 TI - Monitoring tissue oxygen availability with near infrared spectroscopy (NIRS) in health and disease. AB - Near infrared spectroscopy (NIRS) is becoming a widely used research instrument to measure tissue oxygen (O2) status non-invasively. Continuous-wave spectrometers are the most commonly used devices, which provide semi-quantitative changes in oxygenated and deoxygenated hemoglobin in small blood vessels (arterioles, capillaries and venules). Refinement of NIRS hardware and the algorithms used to deconvolute the light absorption signal have improved the resolution and validity of cytochrome oxidase measurements. NIRS has been applied to measure oxygenation in a variety of tissues including muscle, brain and connective tissue, and more recently it has been used in the clinical setting to assess circulatory and metabolic abnormalities. Quantitative measures of blood flow are also possible using NIRS and a light-absorbing tracer, which can be applied to evaluate circulatory responses to exercise along with the assessment of tissue O2 saturation. The venular O2 saturation can be estimated with NIRS by applying venous occlusion and measuring changes in oxygenated vs. total hemoglobin. These various measurements provide the opportunity to evaluate several important metabolic and circulatory patterns in very localized regions of tissue and may be fruitful in the study of occupational syndromes and a variety of diseases. PMID- 11476427 TI - The correlation between peak oxygen uptake (VO2peak) and running performance in children and adolescents. aspects of different units. AB - The method of removing the confounding effect of body mass on peak oxygen uptake simply by dividing the measured values by body mass has been questioned. The aim of the study was to find an expression of body mass for compensating for differences in body mass so as to achieve the best correlation to running performance. In a cross-sectional study, design peak oxygen uptake (VO2peak) was measured in 107 boys in the age range 8-17 years and 88 girls in the age range 8 16 years. The present study indicates that VO2peak do not increase in proportion to body mass during running, rather that VO2peak is inversely related to body mass. When using allometric regression standards with the sexes separated in gender groups, there was a tendency that boys achieved scaling factors closer to unity than did the girls. The increase in subcutaneous fat in the girls throughout the pubertal years is the most likely reason why body mass increased proportionally more than VO2peak in girls. The present study indicates that VO2peak when expressed as milliliters per kilogram per minute (ml kg(-1) min(-1)) is a poor denominator for VO2peak as a function of running performance in children and adolescents. PMID- 11476428 TI - Reliability of sports related functional tests with emphasis on speed and agility in young athletes. AB - The aim of the present investigation was to test the reliability of two sports related functional tests, a speed test (slalom-test) and an agility test (hurdle test). Eleven athletes aged 11 years (8 boys, 3 girls) participated voluntarily in the study. All subjects completed four different test sessions for both the slalom-test and the hurdle-test using six standard track hurdles placed at 2-m intervals along a 12-m length of track. There were no significant differences between testing sessions for either the slalom-test (P=0.99) or the hurdle-test (P=0.96), showing no systematic variation between test times. The intraclass correlation coefficients were 0.96 and 0.90 respectively, indicating a good reliability. We conclude that the slalom-test and the hurdle-test are reliable sports related functional tests for measuring speed and agility in groups of young athletic individuals. PMID- 11476429 TI - Prevalence of knee instability in relation to sports activity. AB - The objective was to estimate the prevalence of knee instability among active athletes and to investigate potential associations to type, amount and duration of sports participation. Based on a questionnaire, 339 athletes provided information about different features of occupation, sports activity and knee instability. The 12-month period prevalence of knee instability and constant or recurrent knee instability, and absence from sport and absence from work due to knee instability, was 22%, 14%, 5% and 1%, respectively. Knee instability as such, and constant or recurrent knee instability were found to be positively associated with female gender and different features of occupational work. In conclusion, knee instability is a commonly reported phenomenon among active athletes. It was found to be independent of the type and the amount of sports activity but highly dependent on female gender, type and amount of occupational work. PMID- 11476430 TI - Physical exercise and musculoskeletal pain among forest industry workers. AB - The study investigated the relations between physical exercise and musculoskeletal pain among forest industry workers. We studied a population of 3312 Finnish forest industry workers, who replied to a questionnaire survey in 1994 (response rate 77%). The outcome variables in this cross-sectional study were the number of days with pain in the low back, neck, shoulder and knee during the preceding 12 months. Multivariable logistic regression models were used in statistical analyses. Active walkers had more sciatic pain, active volleyball players had more shoulder pain and those who practiced trekking actively had more knee pain than those who practiced these activities less. The risk of shoulder pain was more than three times higher for those who played volleyball actively compared to those who played less. In addition, age, mental stress and work related physical loading were strongly associated with musculoskeletal pain. When studying the relations between physical exercise and musculoskeletal pain in a working population, it is important to not only detect the general physical activity but also to specify the different modes of exercise. In addition, the other factors which are strongly related to pain (such as mental stress and work related physical loading) should be taken into account. PMID- 11476431 TI - Is the ulnar nerve damaged in 'handball goalie's elbow'? AB - 'Handball goalie's elbow' has been defined as pain in the elbow region due to repetitive forced hyperextensions of the elbow. Goalkeepers are the players most often suffering from hyperextension trauma to the elbow in European team handball. They may complain of radiating pain or numbness in the ulnar aspect of the forearm in addition to local pain in the elbow region. To detect any injury to the ulnar nerve that could explain the symptoms, we performed a neurological and neurophysiological study in goalkeepers with elbow pain. Nine goalkeepers, with a total of 15 'handball goalie's elbow', were included in this study. Neurological examination revealed a probable ulnar nerve lesion in one player. Neurophysiological and electromyographic examinations (10 examinations) were, however, normal in all players. Handball goalkeepers with elbow problems may suffer from symptoms suggestive of ulnar nerve affection, but serious or permanent injury to the ulnar nerve with wasting or paresis is unusual. PMID- 11476432 TI - Traumatic arthritis in sport. AB - After the introduction of ultrasonography in sports medicine, traumatic arthritis is recognized relatively frequently among athletes. In this case study, the ultrasonographic findings and the prompt effect of ultrasonographically guided intra-articular injection of long-acting steroid are demonstrated in two patients. PMID- 11476433 TI - Extract from the Norwegian National Report on Physical Activity and Health. AB - In 1999 the National Council on Nutrition and Physical Activity in Norway appointed a committee to look into the relationship between health and physical activity, and to provide some guidelines to various populations regarding physical activity. The committee, which consisted of Sigmund B. Stromme, Sigmund A. Anderssen, Ingvar Hjermann, Jorunn Sundgot-Borgen, Sigbjorn Smeland, Sverre Maehlum, and Anita Andaas Aadland, produced a comprehensive 84-page report (rapport nr. 2/2000) on physical activity and health based on the currently available scientific evidence. An extract from the first part of the report, which concentrates on the significance of physical activity for various population groups follows below. PMID- 11476434 TI - Listeria monocytogenes: a causative agent of gastroenteritis? AB - The intestinal tract is the major portal of entry for Listeria monocytogenes, which becomes ingested via contaminated food. The pathogenic strains penetrate the mucosal tissue either directly, via engulfment by enterocytes, or indirectly, via active penetration of the Peyer's patches. There are now several reports clearly demonstrating that, in some cases, acute enteritis may be the only symptom of Listeria infection or may precede the typical symptoms of listeriosis such as sepsis and meningitis or encephalitis. Therefore, Listeria monocytogenes should be included in the list of foodborne gastrointestinal pathogens. Furthermore, a history of enteritis should prompt the physician to include listeriosis, which is often difficult to diagnose, in the differential diagnosis. Although Listeria monocytogenes is undoubtedly a potential enteric pathogen, it is still debatable whether its detection in a routine bacteriological examination of a stool specimen is of clinical significance. PMID- 11476435 TI - Incidence, transmission, and clinical significance of hepatitis G virus infection in hemodialysis patients. AB - A high prevalence of hepatitis G virus (HGV) infection has been noted in patients receiving chronic hemodialysis (HD) therapy, yet the incidence rate and transmission route have rarely been reported. Serum samples from 160 chronically uremic patients in a HD unit were initially collected at the time chronic HD therapy was begun, and thereafter annually in July and, finally, in November 1999. Serum HGV RNA was detected using nested reverse transcription polymerase chain reaction, and HGV E2 antibody was determined using an enzyme immunoassay. Nucleotide sequences of the 5'-noncoding region were studied in the HD patients with HGV viremia. Forty healthy staff members were also enrolled as control subjects. Three of the 40 (7.5%) healthy staff members were positive for HGV RNA or HGV E2 antibodies, in contrast to 40 of the 160 (25%) HD patients, including 14 (8.8%) who were positive for HGV RNA only, 25 (15.6%) who were positive for HGV E2 antibody only, and 1 (0.6%) who had both markers. HGV exposure did not correlate with gender, age, duration of HD therapy, or history of blood transfusions. At least 20 of the 40 (50%) patients with HGV exposure had been infected before the start of chronic HD therapy. Nevertheless, at least nine (22.5%) patients acquired new HGV infections after starting chronic HD therapy, with an incidence rate of > or = 2.6% per year. Three patients with newly acquired HGV viremia after HD therapy was started and two with pre-existing HGV viremia before HD therapy was started had the same nucleotide sequences. HGV and HCV infections (with a prevalence of 14.4%) might have been transmitted independently in HD patients. In addition, HGV infection was not found to cause significant elevation of alanine aminotransferase levels in the group exposed to HGV. To conclude, the incidence of new HGV infections was at least 2.6% per year. In addition to transmission through blood transfusion, HGV may have been transmitted nosocomially patient-to-patient within the HD unit. The compliance with standard universal precautions should be carefully re-examined, but it is not necessary to routinely screen for HGV infection among patients on chronic HD. PMID- 11476436 TI - Long-term infectious complications and their relation to treatment duration in catheter-related Staphylococcus aureus bacteremia. AB - The optimal duration of treatment for catheter-related Staphylococcus aureus bacteremia is not known. Short courses (< or = 2 weeks) of therapy should be viewed with caution because essential data on late complications, such as osteomyelitis and metastatic abscesses, are lacking. This study represents a retrospective analysis of the data from 49 adult patients hospitalised in the period 1994-1996 (mean age, 57 years; range, 20-90 years; 47% male) and from whom Staphylococcus aureus was cultured concomitantly from peripheral blood and catheter segments. Forty-six venous catheters, two arterial catheters, and one unknown type of catheter were used. Forty-four patients were treated with effective anti-Staphylococcus aureus antibiotics. Twenty patients had a favourable outcome, defined as no complication and no death during 1 year of follow-up, 24 patients had complications, 14 patients died due to attributable mortality, and 5 other patients died of an underlying disease without showing signs or symptoms of a complication. Patients were categorised according to the duration of treatment. There were small differences between a shorter (1-14 days) and a longer (>14 days) course of antibiotics with regard to favourable outcome (41% vs. 33%), complications (48% vs. 53%), attributable death (31% vs. 20%), and death due to underlying disease (41% vs. 33%), respectively. The rates of complications and death were high, but a definite conclusion cannot be drawn because the study was underpowered. More randomised trials are needed, but, until the results of such trials are available, the duration of therapy should not be shortened to less than 14 days. PMID- 11476437 TI - Influence of the response to induction therapy on the rate of progression of cytomegalovirus retinitis in AIDS patients on intravitreal maintenance therapy. AB - In order to assess whether complete inactivation of retinitis at the end of induction therapy leads to delayed progression during maintenance therapy with weekly intravitreal ganciclovir, the time to the first progression to retinitis was evaluated in 27 AIDS patients (34 eyes) with stable cytomegalovirus retinitis. Data were censored before the introduction of protease inhibitors. Overall, retinitis progressed in 22 of 34 eyes in a median time of 12 weeks (mean +/- SD, 33 +/- 9 weeks). However, retinitis progressed in 15 of 19 eyes in which only partial inactivation was achieved following induction therapy (median time, 10 weeks; mean +/- SD, 17 +/- 4 weeks) but in only 7 of 15 eyes when complete inactivation was obtained (median time, 59 weeks; mean +/- SD, 56 +/- 19 weeks) (P= 0.02). There were no differences between the groups in CD4+ cell counts, drugs, route of induction treatment, or length of induction therapy. Induction therapy should be prolonged until complete inactivation of retinitis is obtained, since achieving only a partial response appears to be a factor in earlier progression when patients are switched to maintenance therapy with intravitreal ganciclovir. PMID- 11476438 TI - Detection of Giardia lamblia and Entamoeba histolytica in stool samples by two enzyme immunoassays. AB - Two commercially produced enzyme immunoassays (EIAs) to detect antigens of Giardia lamblia and Entamoeba histolytica in stool specimens were evaluated. A total of 276 stool specimens were collected from patients who presented with various medical complaints in the outpatient clinic of the Department of Infectious Diseases and Tropical Medicine, University of Munich. Every specimen was examined by conventional microscopy and tested by both EIA kits. When microscopy was used as the reference standard, the EIA kit detecting Giardia lamblia showed a sensitivity of 100% and a specificity of 99.6%. The EIA kit detecting Entamoeba histolytica had a sensitivity of 81.8% and a specificity of 99.2%. Both tests showed no cross-reactivity with other intestinal protozoa. Antigen detection by EIA has the potential to become a valuable tool capable of making stool diagnostics more effective, although it should not be considered as a replacement for microscopic examination, since other potential pathogens could otherwise escape detection. PMID- 11476439 TI - Serodiagnosis of cat scratch disease: response to Bartonella henselae in children and a review of diagnostic methods. AB - In this study, sera from 116 children without clinical evidence of cat scratch disease (CSD) and sera from 19 children with CSD were investigated by two different immunofluorescent assays (test A and test B) to evaluate the seroprevalence of Bartonella henselae antibodies in this population. Antibodies against Bartonella henselae were found in noninfected children only in low titers: 13% and 3.5% of serum samples were positive by test A and test B, respectively. IgG titers as determined by test A ranged from 1:64 to 1:256, whereas test B did not yield titers >1:64. Of the 19 children with CSD, 7 (37%) and 5 (26%) had low antibody titers (1:64-1:256) on admission by test A and test B, respectively. In such cases, IgM antibodies against Bartonella henselae and/or a titer increase in the following weeks are required to prove suspected CSD. The results indicate that the seroprevalence of Bartonella henselae antibodies in children without evidence of CSD is low, and that the results may differ depending on the tests being used. Low antibody titers, however, were found not only in healthy children but also among patients with CSD, which could be indicative of the beginning or of the end of an illness. Currently, only few data are available concerning the seroprevalence of antibodies to Bartonella spp. among children. Although children are often affected by CSD, the specificity of most tests has been evaluated by investigating blood donors, who are usually adults. Several different serological tests are used for the diagnosis of CSD, including the immunofluorescence assay, the enzyme-linked immunosorbent assay, and Western blot (WB) analysis. The sensitivities of different IFAs range from 14 to 100%, depending on the antigen used, the cut-off chosen, and the test procedures. The current diagnostic value of different serological tests for diagnosis of CSD is reviewed. PMID- 11476440 TI - Immunorestitution diseases in patients not infected with HIV. AB - The aim of this study was to assess the clinical spectrum of immunorestitution disease (IRD) in hospitalized patients over a 12-month period. In nine of 18 patients who presented with clinical deterioration during reduction or cessation of immunosuppressants (n = 6) or bone marrow engraftment (n = 3), IRD cases included the following infections: scabies infestation (n = 1); gastric strongyloidiasis (n = 1); hepatosplenic candidiasis (n = 1); methicillin resistant Staphylococcus aureus abscess formation (n = 2); polyomavirus-related hemorrhagic cystitis (n = 3); and influenza A pneumonitis (n = 1). Immunopathological damage during withdrawal of immunosuppression is an incidental way to uncover an asymptomatic infectious disease. Serial monitoring of hematological and clinical profiles is essential in making a diagnosis of IRD. PMID- 11476441 TI - Survey of staphylococcal enterotoxin genes, exfoliative toxin genes, and toxic shock syndrome toxin 1 gene in non-Staphylococcus aureus species. AB - The aim of this study was to analyze the presence of staphylococcal exotoxin genes in staphylococci other than Staphylococcus aureus. Whereas for Staphylococcus aureus a large spectrum of different exotoxins responsible for toxin-mediated diseases has been described, only few and conflicting data are available regarding toxin production in all other staphylococcal species. A collection of clinical non-Staphylococcus aureus staphylococcal isolates were systematically screened for the presence of genes for toxic shock syndrome toxin 1, exfoliative toxins, and classical enterotoxins by using multiplex polymerase chain reaction enzyme immunoassays. In a total of 461 isolates of 18 different (sub)species derived from clinical specimens, no toxin gene sequences were amplified. The results indicate that the occurrence of toxin genes involved in staphylococcal toxin-mediated diseases is at least very rare in human-derived isolates of non-Staphylococcus aureus species. PMID- 11476442 TI - Lack of change in the distribution of AIDS-defining opportunistic diseases and the related degree of immunodeficiency during the periods before and after the introduction of highly active antiretroviral therapy. AB - In a retrospective study of 86 patients with AIDS-defining diseases diagnosed from 1997 to June 2000, i.e. following consolidated and large-scale introduction of highly active antiretroviral therapy (HAART), and 342 control patients diagnosed with AIDS from 1985 to 1995, i.e. the decade preceding the HAART era, the etiological spectrum and the related level of immunodeficiency of each individual AIDS-defining opportunistic infection were compared. The results show that, despite the progress attained in the management of HIV infection since the introduction of HAART, clinicians facing newly diagnosed cases of AIDS will likely encounter a spectrum of opportunistic diseases and a level of underlying immunodeficiency similar to those observed during the decade preceding the introduction of HAART. Even in recent years, the large majority of patients newly diagnosed with AIDS have not been able to take advantage of HAART prior to diagnosis of their disease. PMID- 11476443 TI - Prosthetic vascular graft infection due to Aspergillus species: case report and literature review. AB - A case of Aspergillus prosthetic graft infection is reported here, accompanied by a review of the literature on this topic. The literature search revealed only 13 other cases reported to date. This infection is usually acquired through contamination at the time of surgery and affects immunocompetent patients. Aspergillus fumigatus is the causative species in most cases. Remarkably, fever is absent in about one-half of all cases, and blood cultures are usually negative. Concomitant vertebral osteomyelitis is commonly observed when the aorta is involved. Cure of this serious infection may be achieved with antifungal therapy, excision of the infected graft and extra-anatomic bypass. PMID- 11476444 TI - Implications of the simultaneous presence of metronidazole-susceptible and resistant Helicobacter pylori colonies within a single biopsy specimen. AB - This study examined whether the simultaneous presence of metronidazole susceptible and -resistant Helicobacter pylori colonies in a single biopsy specimen is caused by a multiple strain infection with a susceptible and a resistant strain or by two subpopulations within a single strain. Single colonies obtained from seven biopsy specimens known to harbour both susceptible and resistant Helicobacter pylori were fingerprinted by restricted fragment length polymorphism typing of the ureC gene and by the random amplified polymorphic DNA procedure. Metronidazole susceptibility was determined by the E test. The results indicated that the occurrence of metronidazole-resistant and metronidazole susceptible bacteria within a single biopsy does not imply the presence of a multiple strain infection with one resistant and one sensitive strain. PMID- 11476445 TI - Competitive polymerase chain reaction used to diagnose cutaneous leishmaniasis in German soldiers infected during military exercises in French Guiana. AB - A competitive polymerase chain reaction (PCR) followed by enzyme-linked immunoassay-based verification of PCR products has been developed, which facilitated the diagnosis of leishmaniasis in two German soldiers who underwent survival training in the jungle of French Guiana and returned with therapy resistant pyoderma-like lesions. After treatment with liposomal amphotericin B, the skin manifestations disappeared, and leishmania DNA could no longer be detected by PCR. In the context of growing military involvement in areas where leishmaniasis is prevalent, this assay may help detect or, due to its internal controls, exclude cases of infection with this parasite. PMID- 11476446 TI - Antibiotic susceptibility and phage typing of methicillin-resistant and sensitive Staphylococcus aureus clinical isolates at three periods during 1991 1997. AB - The aim of the study presented here was to evaluate the antibiotic susceptibility of Staphylococcus aureus over a 7-year period. A total of 2,122 clinical isolates of Staphylococcus aureus were collected from hospitalized patients at 3-year intervals during the period 1991-1997. The prevalence of methicillin-resistant isolates was 41.6%, 38.2% and 36% in 1991, 1994 and 1997, respectively; all of these isolates were sensitive to vancomycin. Over the study period, resistance to pristinamycin and fusidic acid increased slightly and resistance to imipenem, rifampicin and amikacin increased greatly, while resistance to trimethoprim/sulfamethoxazole decreased. For methicillin-sensitive Staphylococcus aureus isolates, significantly increased resistance was observed against amikacin only. Phage typing was conducted using the international set of phages. All of the isolates that were sensitive to group I, group II, or group V phages were sensitive to methicillin. Of the isolates that were sensitive to group III phages, 96% were methicillin resistant, and 70.5% of them were sensitive to phages 75 and 85. PMID- 11476447 TI - Clinical manifestations and predictors of survival in AIDS patients with disseminated Mycobacterium avium infection. PMID- 11476448 TI - Verocytotoxin-producing Escherichia coli serotype O26:H11 outbreak in an Irish creche. PMID- 11476449 TI - Successful treatment of vancomycin-resistant Enterococcus meningitis with linezolid. PMID- 11476450 TI - Severe ciprofloxacin-induced acute hepatitis. PMID- 11476451 TI - Haemophilus influenzae invasive disease in Italy, 1997-1998. PMID- 11476452 TI - Group A streptococcal sepsis secondary to peritonitis and acute pelvic inflammatory disease. PMID- 11476453 TI - Aspergillus flavus infection of an aortic bypass. PMID- 11476454 TI - Dysphonia and dysphagia as primary manifestations of invasive aspergillosis. PMID- 11476455 TI - No prescription needed--no doctor either. PMID- 11476456 TI - Safety of fexofenadine in children treated for seasonal allergic rhinitis. AB - BACKGROUND: The incidence of allergic rhinitis in children is increasing. OBJECTIVE: To evaluate the safety of fexofenadine HCI in children ages 6 through 11 years for treatment of seasonal allergic rhinitis. METHODS: Two large, double blind, randomized, placebo-controlled, parallel studies with identical protocols included patients with a positive skin test to fall allergen(s) and allergic rhinitis symptoms. Patients were randomized to receive fexofenadine 15, 30, or 60 mg or placebo twice daily for 2 weeks after a 1-week placebo lead-in. Safety was evaluated through adverse event reporting, electrocardiograms, and pre- and posttreatment laboratory panels and physical examinations. RESULTS: A total of 875 patients from both studies were eligible for safety analyses. Ten patients (5 on placebo, 5 on fexofenadine) discontinued because of an adverse event; no event that resulted in discontinuation was judged to be caused by study medication. Incidence of adverse events was similar in active and placebo groups, and did not increase with increasing fexofenadine dose: 36.2% (83 of 229) in the placebo group versus 35.3% (79 of 224), 36.8% (77 of 209), and 34.7% (74 of 213) in the 15, 30, and 60 mg twice-daily fexofenadine groups, respectively. Headache was the most commonly reported adverse event (6.6%, 8.0%, 7.2%, and 9.4% in the placebo, 15, 30, 60 mg twice-daily fexofenadine groups, respectively). Clinical, vital sign, electrocardiogram, and laboratory measures were similar in active and placebo groups. There was no statistically significant mean change from baseline in any electrocardiogram parameter after fexofenadine treatment. CONCLUSIONS: Fexofenadine, 15, 30, and 60 mg twice daily, was safe and well tolerated in this large pediatric patient population. PMID- 11476457 TI - Clinical features of acute anaphylaxis in patients admitted to a university hospital: an 11-year retrospective review (1985-1996). AB - BACKGROUND: Although anaphylaxis is considered a life-threatening event, there is a lack of information on the clinical characteristics at presentation, both in adults and in children. OBJECTIVE: To describe in a nonselected population the clinical characteristics and the treatments of acute anaphylaxis triggered by different agents. METHODS: This is a retrospective review of the clinical features of 113 episodes of acute anaphylaxis resulting in admission to a university hospital. Initially, the 107 patients visited the emergency room and were then admitted to the hospital. RESULTS: Most anaphylactic events (63%) occurred at home. The most frequent symptoms involved the respiratory system (78%) and the skin (90%). Drugs, especially nonsteroidal anti-inflammatory drugs and antibiotics, were the most frequent cause of anaphylaxis in adults (49%). Patients with drug-induced anaphylaxis were older and more often had cardiovascular symptoms (hypotension and tachycardia) (P = 0.0064). Hymenoptera venom was the second most frequent cause of anaphylaxis (29%). Most of the patients with hymenoptera venom anaphylaxis were male (80%) and more frequently they had no history of atopy (P = 0.012). In food-induced anaphylaxis, the cardiovascular system was less likely to be involved (P < 0.05) (39%). Seafood seems to be frequently involved in food-induced anaphylaxis in our area. Specific immunotherapy-induced anaphylaxis occurred more often in younger patients (P = 0.032). Epinephrine seems to be underused in Italy (only 15% of patients received it), especially for respiratory symptoms. CONCLUSIONS: Anaphylaxis triggered by different agents may have different clinical presentations and may occur in different types of patients. In Italy, the inadequate use of epinephrine for anaphylaxis treatment needs to be publicized to both physicians and the general population. PMID- 11476458 TI - FDA panel recommends OTC status for second-generation antihistamines. PMID- 11476459 TI - Age-dependent relationship between bronchial hyperresponsiveness to methacholine and total serum IgE level in asthmatic children. AB - BACKGROUND: A relationship between nonspecific bronchial hyperresponsiveness and allergic airway inflammation has been reported in children and in adults with asthma, but the relationship in infants with asthma is still unclear. OBJECTIVE: To evaluate the relationship between bronchial hyperresponsiveness and total serum IgE level throughout childhood. Bronchial reactivity to methacholine from the age of 1 to 16 years was studied by methacholine inhalation challenge using transcutaneous oxygen pressure (tcPO2) monitoring. METHODS: Two hundred one asthmatic children (boys:girls = 132:69; 7.3+/-4.0 years of age, mean +/- SD) were enrolled in this study. The tcPO2 was measured using a tcPO2 monitor. Serial doses of methacholine were doubled until a 10% decrease in tcPO2 from the baseline was reached. The cumulative dose of methacholine at the inflection point of tcPO2 was considered to represent the bronchial reactivity to methacholine. RESULTS: There was no relationship between the cumulative dose of methacholine at the inflection point of tcPO2 and total serum IgE level in the group of children aged 1 to 4 years (P = 0.212), but significant correlations were found in the groups aged 5 to 10 years and 11 to 16 years (P = 0.044 and P = 0.014, respectively). CONCLUSIONS: We conclude that there is an age-dependent relationship between bronchial reactivity to methacholine and the total serum IgE level and that inhaled allergens, which were more common allergens in older children, may have some effects on the degree of bronchial reactivity to methacholine in children with asthma. PMID- 11476460 TI - Pinus pollen aerobiology and clinical sensitization in northwest Spain. AB - BACKGROUND: Pinus pollen allergy has been generally considered to be rare and clinically insignificant. In our geographical area, Pinus pollen constitutes one of the most predominant pollen together with Poaceae and Urticaceae pollen. Pinus pinaster and Pinus radiata are the main species. OBJECTIVE: This work aimed to determine the atmospheric fluctuations and the existence of patients monosensitized to Pinus pollen in our region. METHODS: Patients attending our outpatient clinic in the last 4 years with positive cutaneous tests to Pinus pollen and with respiratory symptoms were selected. They were skin tested with commercial extracts of a battery of inhalants including Pinus pollen. Serologic specific IgE to Pinus pollen was performed by CAP system (Pharmacia & Upjohn, Uppsala, Sweden). Airborne Pinus pollen counts in our city were obtained for a 5 year period (1995 to 1999). RESULTS: We present 10 patients with sensitivity to Pinus pollen and with symptomatology coinciding with Pinus pollen season (February to April). Most of these patients were monosensitizated to Pinus pollen and suffered from seasonal rhinoconjunctivitis. Pinus pollen is present in our area in large amounts from February to April with a peak pollen count in March and April. CONCLUSIONS: Pinus pollen may be an important allergen since by itself can be the only cause of allergic disease. Therefore, we think that it must be taken into account in patients living in areas with high Pinus pollen concentrations and with seasonal respiratory disease. PMID- 11476461 TI - Efficacy and safety of long-term treatment of asthmatic patients with pranlukast, a cysteinyl-leukotriene-receptor antagonist: four-year followup study. AB - BACKGROUND: There are few studies that have examined the long-term efficacy and safety of pranlukast, a leukotriene receptor antagonist, in asthmatic patients. METHODS: Sixty-three asthmatic patients were entered in this 4-year study [group 1, mild or moderate (N = 22); group 2, severe without using oral prednisolone (N = 22); group 3, severe with using oral prednisolone (N = 19)]. Pranlukast was administered at 225 mg twice daily to 14 subjects in group 1 (group 1p), 14 in group 2 (group 2p), and 11 in group 3 (group 3p), chosen for pranlukast additional therapy at random. Another group of 24 asthmatic patients was assigned to conventional therapy group (groups 1c, 2c, and 3c). Efficacy was determined by improvement in symptom score, peak expiratory flow rate (PEFR) percentage predicted, reduced daily variability of PEFR (percentage), and reduced frequency of use of rescue beta2-agonist (times per week). RESULTS: In groups 1p and 2p, PEFR percentage predicted began to improve from 2 weeks after commencement of treatment. The symptom score, daily variability of PEFR, and use of rescue beta2 agonist diminished significantly. In group 3p, pranlukast was ineffective in improving PEFR percentage predicted. All but two patients continued to receive pranlukast and no adverse effects were noted, at least during the 16-week therapy. Further, 22 patients continued to receive pranlukast for 4 years, and none experienced any adverse effects. CONCLUSIONS: We showed in this study that long-term treatment with pranlukast is effective for asthmatic patients without any adverse effects. PMID- 11476462 TI - Double-blind multicenter study on the efficacy and tolerability of cetirizine compared with oxatomide in chronic idiopathic urticaria in preschool children. AB - BACKGROUND: There are no studies on the use of cetirizine in children under the age of 6. OBJECTIVE: We compared the efficacy and tolerability of cetirizine in patients with idiopathic chronic urticaria to the more widely used antihistamine, oxatomide. METHODS: This double-blind study was performed on 62 patients (38 male and 24 female) with idiopathic chronic urticaria, recruited from four different medical centers of the national territory (Ancona, Cagliari, Catania, and Messina). The children's ages ranged from 2 to 6 years (mean 3.85). The patients were randomly assigned to two treatment groups: one group treated 31 children with cetirizine at a dosage of 5 mg q.d., and a second group treated 31 children for the same amount of time with oxatomide, at a dosage of 25 mg q.d. Sixty-two children began the treatment, but five did not finish the study (three in the cetirizine and two in the oxatomide group). Thus, the clinical study and the statistical evaluation were conducted on 57 children (28 cetirizine and 29 oxatomide). The Student's t test was used to compare severity of the illness and changes in the hematochemical tests. RESULTS: Overall, the effectiveness of the two medications in treating erythema, papules, edema, and itching showed comparable therapeutic activity (P < 0.001). Neither medication produced significant side effects. CONCLUSIONS: The results of the present study suggest that cetirizine may represent an effective and safe pharmacologic therapy for chronic urticaria in preschool children. There was no evidence for changes in hematochemical and urinary values, demonstrating the safety and the tolerability of the two antihistamines, even when given to young children. PMID- 11476463 TI - Anaphylaxis associated with antiphospholipid syndrome. AB - BACKGROUND: To our knowledge, no previously published reports have described food induced anaphylaxis associated with the antiphospholipid syndrome. OBJECTIVE: We undertook a study of four patients with thrombosis associated with the antiphospholipid syndrome after each patient experienced anaphylaxis attributable to ingestion of vegetal foods. METHODS: IgE antibody levels to various foods were determined in serum specimens from the study patients, and skin prick tests with the same allergens were conducted to determine their in vivo responses. Hematologic, cardiopulmonary, vascular, and rheumatologic studies were also performed. IgG anticardiolipin antibody levels were determined by ELISA. RESULTS: All four patients fulfilled the criteria for antiphospholipid syndrome and had high levels of specific IgE antibodies for certain food allergens. By immunoblot analysis, the presence of serum IgE specific for a 45-kD protein band in an almond extract was detected in these four patients who experienced food-related anaphylaxis. No specific IgE was detected in sera from normal subjects. No IgE antibodies specific for the food panallergen lipid transfer proteins were detected. CONCLUSIONS: This is the first report of severe food-precipitated anaphylaxis associated with the antiphospholipid syndrome and the first description of a patient with allergy to blackberry. The possible involvement of food panallergens distinct from lipid transfer proteins is also discussed. PMID- 11476464 TI - The many faces of systemic mastocytosis. AB - OBJECTIVE: This short review article will augment the reader's knowledge of mast cell physiology and will offer an overview of current information on the pathophysiology, heterogeneity, and treatment of human mastocystosis. DATA SOURCES AND STUDY SELECTION: Articles published since 1980, textbooks, information from computerized databases, references identified from bibliographies of relevant articles, and books published in the last 10 years. RESULTS AND CONCLUSIONS: Mastocytosis is a complex disease with a multitude of clinical presentations, often misdiagnosed, which can embrace characteristics of other diseases and generate a chameleon-like picture. Mast cells possess many important physiologic functions in the human body, but as a consequence of poorly understood events, they can also start a cascade of pathologic reactions. Although a great deal is known about mechanisms involved in physiologic and pathologic processes of mast cells, many areas are waiting to be explored in this millennium. PMID- 11476465 TI - Cockroach allergen abatement with extermination and sodium hypochlorite cleaning in inner-city homes. AB - BACKGROUND: Although the importance of cockroach allergen in chronic asthma has now been well defined, little progress has been made in the control of cockroach allergen in infested homes. OBJECTIVE: The objective of this study was to examine the ability of a combination of professional pest extermination and household cleaning using a solution of sodium hypochlorite to reduce cockroach infestation and allergen levels in cockroach infested homes. METHODS: Seventeen cockroach infested homes were studied with three homes serving as controls. In the intervention homes, a professional exterminator applied 0.05% abamectin twice at 2-week intervals at study entry and a professional cleaner cleaned the homes before and after the extermination. All washable surfaces were cleaned throughout the study with a solution of 0.5% hypochlorite. Monthly home visits were conducted to inspect the home, interview the homeowner, place passive cockroach traps, and to collect settled dust samples from the kitchen, bedroom, and TV/living room. RESULTS: The number of cockroaches in the passive traps decreased rapidly after the initial intervention in most homes. Median Blatella germanica allergen 1 levels in the settled dust samples fell by 91% in the kitchen, 78% in the bedroom, and 77% in the living room over the course of the study in the intervention homes but gradually rose in the control homes. The overall reductions were very similar to those seen in a previous study with a similar protocol except for the use of the sodium hypochlorite. CONCLUSIONS: Successful extermination is possible in most inner-city homes and cockroach allergen levels can be reduced by 80% to 90%. However, 0.5% sodium hypochlorite did not seem to improve allergen reduction, and in many homes, allergen levels remained above the proposed threshold of 8 U/g of dust throughout the study. PMID- 11476466 TI - A case of allergy to beer showing cross-reactivity between lipid transfer proteins. AB - BACKGROUND: Lipid transfer proteins (LTPs) are highly conserved proteins present in a broad spectrum of fruits and vegetables that might represent a novel plant panallergen. OBJECTIVE: To demonstrate that LTP is an important allergen in beer and that beer LTP cross-reacts with LTP from botanically unrelated plant-derived foods. METHODS: Serum from a patient with clinical allergy to both beer and Rosaceae was studied for IgE reactivity to LTP to several vegetable foods by RAST, ELISA, immunoblot, and inhibition studies. RESULTS: Patient's serum showed a strong IgE reactivity to LTP purified from peach peel, carrot, and broccoli, and to a 10 kD protein in both apple and peach immunoblots, whereas no reactivity to birch cross-reactive allergens such as Bet v 1, profilin, or carbohydrates was found. In inhibition studies, preabsorption of serum with apple, walnut, hazelnut, peanut, corn, and rice caused a fall of 97%, 20%, 66%, 91%, 94%, and 93%, respectively, of its reactivity to peach LTP. Beer RAST fell from 1.8 IU/mL to <0.1 IU/mL when a patient's serum was preabsorbed with recombinant carrot LTP. CONCLUSIONS: LTP is a relevant allergen in beer. Beer LTP may cross-react with LTP from several other plant-derived foods. PMID- 11476467 TI - Allergy to nonspecific lipid transfer proteins in Rosaceae: a comparative study of different in vivo diagnostic methods. AB - BACKGROUND: Lipid transfer proteins (LTPs) are the major allergens in patients sensitive to Rosaceae (apple, peach, apricot, cherry, plum, and pear) who are not allergic to birch pollen. OBJECTIVE: The purpose of this study was to find a sensitive, specific, and relatively easy method for detection of LTP-sensitive patients. METHODS: We studied 36 persons who experienced oral allergy syndrome after the ingestion of fruits in the family Rosaceae. This study cohort was divided into two groups: 18 without allergy to birch pollen (patients) and 18 with birch pollen allergy (control subjects). All were tested by skin prick tests (SPTs) with fresh Golden Delicious apple, fresh peach, and extracts of peel and pulp from both fruits. Their specific IgE reactivities against peach peel extract were further investigated by immunoblot analysis. RESULTS: All 18 subjects in the control group showed strongly positive skin reactions with both fresh apple and fresh peach, whereas no skin reactivity was found with extracts from peach peel, peach pulp, or apple pulp. Extract of apple peel produced positive skin reactions in 17 of 18 control subjects; however, the wheals were generally smaller than those induced by fresh fruits. Immunoblot analysis showed no reactivity for peach peel extract. In contrast, the SPTs with fresh fruits showed that some of the 18 patients had strongly positive reactions, but others had weak reactions or negative responses. Further, in a high proportion of the patients, consecutive SPT with fresh apple yielded inconsistent results. In all patients, SPTs with extracts from apple pulp and peach pulp were negative, whereas SPTs with peel extracts were strongly positive in all patients. In most patients, the wheal area induced by SPT with peel extracts was larger than that induced by SPTs with fresh fruits. Immunoblot analysis showed that serum specimens from all 18 patients reacted with a 10-kD protein in peach peel. This is the molecular mass of LTPs. CONCLUSIONS: In birch pollen-allergic patients, the SPTs with fresh foods still remains the most reliable method of diagnosing vegetable food hypersensitivity. In contrast, in patients not allergic to birch pollen, the most reliable strategy for detection of patients sensitive to LTPs is skin prick testing with properly prepared fruit peel extracts. The loss of Bet v 1- and Bet v 2-like structures, which probably occurs during extraction, may facilitate immediate identification of the relevant allergen. PMID- 11476468 TI - Anaphylaxis to celecoxib. AB - BACKGROUND: Adverse reactions such as urticaria, angioedema, asthma, and anaphylaxis are known to be associated with nonsteroidal anti-inflammatory agents (NSAIDs). Celecoxib (Pfizer/Searle, Caguas, PR) is a new NSAID that differs in structure and mechanism of action of other similar drugs of this class. OBJECTIVE: Evaluation of a case of anaphylaxis to celecoxib (Celebrex). METHODS AND RESULTS: This report describes a 55-year-old woman who experienced the acute onset of pruritus, urticaria, respiratory distress, and hypotension minutes after ingesting a celecoxib capsule. She had taken the drug a previous time for tendonitis without difficulty. Treatment with epinephrine, corticosteroids, and intravenous fluids was successful. An IgE mechanism could not be detected. She has avoided the drug and has had no further problems. CONCLUSIONS: This is the first patient report of anaphylaxis attributable to celecoxib, a new NSAID. This suggests that physicians and other health care professionals should be aware of the potential serious side effects of this drug. PMID- 11476469 TI - Effects of pranlukast on aspirin-induced bronchoconstriction: differences in chemical mediators between aspirin-intolerant and tolerant asthmatic patients. AB - BACKGROUND: Aspirin inhibits cyclooxygenase activity and modifies production of the arachidonate cascade in aspirin-induced asthma. The aim of the present study was to examine the effects of leukotriene (LT) receptor antagonist on aspirin challenge on eosinophil activity and chemical mediators released into the airway of asthmatic patients. METHODS: Aspirin oral provocation test was performed in aspirin-intolerant asthmatic patients (AIA; N = 7) and aspirin-tolerant asthmatic patients (ATA; N = 7). In AIA, LT receptor antagonist (pranlukast) was administered orally 2 hours before the test, and its inhibitory effects on sputum LTC4+C4, eosinophil cationic protein (ECP), eosinophil count, urinary LTE4/creatinine (Cr), 11-dehydrothromboxane (11-dhTX) B2/Cr, serum LTC4+D4, ECP, and peripheral blood eosinophil count were compared with the findings in ATA subjects. RESULTS: In AIA, aspirin induced an immediate reaction associated with increased urinary LTE4/Cr and sputum ECP and a fall in urinary 11-dhTXB2/Cr. Pranlukast inhibited the bronchial reaction and an increase in sputum ECP after threshold dosed of ASA, but failed to change aspirin-induced LT production in sputum and urine. In ATA, aspirin challenge was only associated with a fall in urinary 11-dhTXB2. CONCLUSIONS: Our results indicated that aspirin-induced asthma is associated with overproduction of LT with a shift to the 5-lipoxygenase series of the arachidonate cascade and that leukotriene receptor antagonist are useful for AIA through inhibition of production of LT and eosinophilic inflammation in the airway. PMID- 11476470 TI - The consideration of immunotherapy in the treatment of allergic asthma. AB - BACKGROUND: Immunotherapy (IT) has undergone rigorous trials to evaluate its therapeutic benefit in the treatment of allergic respiratory disease. The tools of molecular biology have provided a framework with which to begin to understand the mechanistic effects of IT on the underlying inflammatory component of allergic respiratory disease. RESULTS: The clinical relevance of these observations belies our understanding of allergic inflammation as the subsoil for the development of abnormal airway physiology, heightened bronchial reactivity, and the development of chronic asthmatic symptomatology. CONCLUSIONS: IT provides the potential to downregulate this inflammatory cascade, reduce IgE antibody production, and attenuate symptoms. Conceptually, early intervention of allergic disease holds the most promise as a therapeutic intervention capable of arresting the progression of the disease, altering the severity of the disease, and/or preventing the development of the respiratory disease process. PMID- 11476471 TI - Immunotherapy for asthma: unfavorable studies. AB - INTRODUCTION: Many studies have demonstrated efficacy of immunotherapy (IT) for asthma. Two meta-analyses reached the same conclusion. Because some studies failed to demonstrate efficacy, it is instructive to review reasons why that may have been the case. METHODS: Two meta-analyses were used to identify studies that did and did not demonstrate efficacy. These were reviewed to identify the sources of variability. RESULTS: Most studies are able to demonstrate a treatment effect of IT although they have wide confidence intervals. The studies were heterogeneous with respect to selection of subjects, underlying populations, the main treatment protocol, concomitant treatments, the duration of treatment and followup, and the effect of the protocol on concurrent treatment. There were also problems with the blinding. In most cases, it was difficult to determine how the doses of extract given compared between studies. CONCLUSIONS: Studies that fail to demonstrate a treatment effect of IT may do so for many reasons. Given the heterogeneity of IT clinical trials as a group, it is not surprising that variable results occur. The use of multiple studies in a meta-analysis with its underlying assumption of homogeneity is problematic. PMID- 11476472 TI - Immunotherapy historical perspective. AB - OBJECTIVE: To review early history of specific immunotherapy, particularly the contribution of Robert A. Cooke. DATA SOURCES: Review of literature. CONCLUSIONS: Cooke was an important investigator developing the basic method of immunotherapy as it is practiced today. His contributions to the field of allergy are staggering. They include the elucidation of blocking antibodies, passive transfers, dosage techniques, testing techniques, and more. It is our hope this consensus conference will lead to more national and international collaborative efforts focusing on different subtopics of allergen immunotherapy. We intend to bring cutting edge information, discussions, and consensus agreements for our own members, for other physicians, and for the public at large. PMID- 11476473 TI - Analyzing meta-analyses of specific immunotherapy in the treatment of asthma. AB - OBJECTIVE: To review the differences and similarities of the meta-analyses dealing with asthma immunotherapy (IT). DATA SOURCES: Published papers by Abramson et al (Am J Respir Crit Care Med 1995;151:969-974) and Ross et al (Clin Ther 2000;22:329-341). RESULTS: Based on the review of published data concerning meta-analyses of published clinical studies using IT to treat asthma, it is an effective treatment. IT can significantly reduce asthma symptoms and medication requirements. Both specific and nonspecific bronchial hyperresponsiveness were decreased, but improvement as measured by lung function tests was inconsistent. CONCLUSIONS: IT needs to be considered in the treatment of allergic asthmatic patients. PMID- 11476474 TI - Specific immunotherapy in rhinitis and asthma. AB - OBJECTIVE: The primary objective of this review is to discuss the efficacy of immunotherapy on combined lung and nasal symptoms, as well as data suggesting that allergen-specific immunotherapy (SIT) may, by altering the natural course of allergic disease, have a preventive effect on rhinitis and asthma. DATA SOURCES: A review of literature published on allergen-specific immunotherapy in the treatment of allergic rhinitis and asthma was performed. The review represents a synthesis of these sources and the expert opinion of the author. STUDY SELECTION: The expert opinion of the authors was used to select the relevant data for the review. RESULTS: Epidemiologic studies have consistently shown that asthma and rhinitis often co-exist in the same patients, and rhinitis often precedes the onset of asthma. SIT has been shown to be effective as a curative treatment for these allergic diseases, but there is increasing evidence indicating that SIT also may have a preventive effect. A study of children monosensitized to house dust mites showed that immunotherapy altered the natural course of allergy in preventing the development of additional sensitizations. CONCLUSIONS: At present, SIT seems to be the only treatment that may modify the course of the disease, either by preventing the development of new sensitivities or by altering the natural history of asthma. When SIT is introduced to patients with only allergic rhinoconjunctivitis, it may stop the development of asthma. SIT should be started early in the disease process, as soon as allergy has been diagnosed, to modify the spontaneous long-term progress of the allergic inflammation and disease. PMID- 11476475 TI - Preventive aspects of immunotherapy: prevention for children at risk of developing asthma. AB - OBJECTIVES: This review describes the connection between hay fever and asthma in patients suffering from specific IgE-mediated allergy. The short-term and longterm benefits of allergen specific immunotherapy (SIT), including potential prevention of exacerbation of disease from hay fever into asthma, are discussed. DATA SOURCES: A review of literature on recent studies of SIT as treatment for allergic disease was conducted. Preliminary data emerging from the ongoing preventive allergy treatment study in Northern Europe are also presented. STUDY SELECTION: The author's expert opinion was used to select relevant data. RESULTS: Treatment with allergenic vaccines (immunotherapy) resulted in decreased symptoms of allergic rhinitis, decreased development of new allergies, and less asthma in the treated group, compared with the untreated control group. CONCLUSIONS: SIT performed with characterized and standardized allergen extracts in an optimal dosage has been shown to interfere with the basic mechanisms of the allergy, resulting in immediate symptomatic relief and offering the patient a long-lasting and preventive effect. PMID- 11476476 TI - Systemic reactions and fatalities associated with allergen immunotherapy. AB - OBJECTIVE: The primary objective of this review is to discuss systemic allergic reactions and risk factors associated with the injection of allergen vaccines. DATA SOURCES: A review of the literature on anaphylactic reaction, adverse effects, and fatalities associated with allergen immunotherapy (IT) was conducted. STUDY SELECTION: The expert opinion of the author was used to select relevant data. RESULTS: Systemic allergic reactions associated with the injection of allergen vaccines usually begin within 20 minutes. However, on occasion, they begin 20 to 30 minutes or longer after an injection. Such reactions can also occur after allergen skin testing. Most reactions associated with skin testing and allergen IT are mild and readily respond to appropriate treatment. However, severe and even fatal reactions have been reported with both skin testing and IT. CONCLUSIONS: Risk factors for skin testing and allergen IT include: 1) patients, particularly asthmatic patients, suffering with seasonal exacerbation of their symptoms; 2) patients who demonstrate exquisite sensitivity to particular allergen(s); 3) patients on beta-blockers; 4) patients with asthma, especially if their asthma is unstable; 5) patients in whom rush IT is used; and 6) patients in whom high doses of potent standardized allergen vaccines are used. It is essential that strict attention be paid to the risk factors for systemic reactions, and that techniques and management be initiated both before and after skin testing or IT to minimize these risks. Done properly, the risk of skin testing and IT is minimal. PMID- 11476477 TI - Epidemiology and heterogeneity of asthma. AB - LEARNING OBJECTIVES: The primary objective of this review is to discuss the prevalence, risk factors, and natural history of asthma and its relationship to chronic airflow obstruction. DATA SOURCES: A review of PubMed (National Library of Medicine) articles on these topics for the years 1995 through 2000 was performed. In addition, references identified some other review articles, and discussions with experts in the field were included. The review represents a synthesis of these sources and the expert opinion of the author. STUDY SELECTION: The expert opinion of the author was used to select the relevant data for the review. RESULTS: Asthma is a disease that begins in early childhood with more than 90% of cases diagnosed by the age of 6 years. Recall bias for early life events, coupled with growth and lung function, leads to a high prevalence of the intermediate phenotypes of allergy and airways responsiveness in early adult life. Interaction of these intermediate phenotypes with environmental exposures results in recurrent disease which often appears to be incidental to the practitioner, but is actually a recapitulation of early childhood events. CONCLUSIONS: With the development of half a million new cases each year, asthma could be characterized as an epidemic. Increased airways responsiveness and an asthma diagnosis in later adult life are associated with an accelerated decline in lung function and the development of chronic obstructive lung disease. PMID- 11476478 TI - Appropriate and inappropriate use of immunotherapy. AB - OBJECTIVE: The primary objective of this review is to discuss indications for appropriate and inappropriate use of allergen immunotherapy (IT), including discussion of contraindications, adverse events, and alternative protocols and methods. DATA SOURCES: A review of the literature on indications, contraindications, adverse events, and alternative methods and protocols associated with allergen IT was conducted. STUDY SELECTION: The opinion of the author was used to select and review relevant data. RESULTS: Clinical trials have proven the benefit of allergen IT, for allergic rhinitis, asthma, and anaphylaxis because of stinging insects, such as Hymenoptera and fire ants. Allergen IT may improve quality of life and decrease medication requirement for allergic rhinitis and asthma, especially for younger patients, and probably will assist in the prevention of the progression of allergic disease from rhinitis to asthma. Subcutaneous allergen IT is well established as clinically effective through multiple, blinded, placebo-controlled studies. The data for alternate therapies and routes of therapy are extensively reviewed and critiqued. CONCLUSIONS: Allergen IT is a highly valuable form of treatment of IgE-mediated diseases. Documentation of sensitivity to allergen sensitization associated with symptoms is critical before use of allergen IT. Symptoms should also be of sufficient duration and severity to warrant IT. Before allergen IT is used, there must be an available allergen extract that is suitable for treatment of the relative sensitivity. It is essential for patients receiving allergen IT to understand the treatment principle, the frequency of injections, the duration of treatment, the risk and signs of adverse events, the magnitude of the efficacy, and the essential nature of patient compliance. Subcutaneous allergen IT is the only current mode of therapy in the United States that has been shown to be effective. PMID- 11476479 TI - New treatments and the future role of immunotherapy: anti-IgE. AB - OBJECTIVE: The review outlines an overview of anti-IgE, along with a description of the production and mechanism of action of the finished molecule. The immunologic effect of the drug and the drug complexes are detailed, and proof of efficacy is offered. The review also speculates about the role of anti-IgE in clinical use, along with potential future therapies, including gene substitution and genetic alteration. DATA SOURCES: The author reviewed literature studying IgE and anti-IgE antibody. STUDY SELECTION: The expert opinion of the author was used to select relevant data. RESULTS: The injection of anti-IgE results in four proven reactions: free antibody is bound; not bound antibody is not dislodged to any degree; plasma B cell production of IgE drops; and high-affinity Fc epsilon RI and low-affinity Fc epsilon RII on mast cells and basophils are downregulated. CONCLUSIONS: Based on observation of clinical reaction, anti-IgE is truly immunotherapy. Current opinion among clinical investigators about the impact of the impending approval of this new drug is that it will not replace standard immunotherapy. How it will fit into the armamentarium of physicians dealing with allergic disease, and protocols for its use, are still being debated. PMID- 11476480 TI - Asthma: best treatment options. AB - LEARNING OBJECTIVES: To understand the principles of asthma management in patients of different ages, and to outline major avoidance measures, classes of medications available, and indications for immunotherapy (IT). DATA SOURCES: MEDLINE and major textbooks of allergy and immunology. CONCLUSIONS: There has been an understanding of asthma as a chronic inflammatory disease for many decades but recent information about mediators and cytokines has led to new therapies and better understanding of the effects of IT and other preventive measures. The best treatment for any individual patient depends upon many factors and is a decision to be made between the patient and the physician. It seems that the best response to IT occurs in those who are highly sensitive to unavoidable allergens, and who experienced a late-phase asthmatic response to the allergen initially. New data suggest that IT should be considered early, as it may prevent progression of asthma to more severe, less reversible disease. The best therapy results in the ablation of airway early- and late-phase reactivity. PMID- 11476481 TI - Type I polyketide biosynthesis in bacteria (part B). PMID- 11476482 TI - Steroids: reactions and partial synthesis. PMID- 11476483 TI - DNA minor-groove recognition by small molecules. PMID- 11476484 TI - Anthocyanins and other flavonoids. PMID- 11476485 TI - The biosynthesis of shikimate metabolites. PMID- 11476487 TI - Aerosol chemical and optical properties during the Mt. Zirkel Visibility Study. AB - Aerosol chemical and optical properties were measured near the Mt. Zirkel Wilderness Area in northwestern Colorado. Six-hour PM2.5 (particles with aerodynamic diameters less than 2.5 microm) mass concentrations and PM2.5 dry particle light scattering at 550 nm averaged 4.6 microg m(-3) and 8.6 Mm(-1), respectively. Sulfates, organic carbon, and geological material were the principle components of particle mass and light scattering. Hygroscopic growth was consistent with that expected for ammonium sulfate aerosols. Size distributions derived from three-wavelength (i.e., 450, 550, and 700 nm) nephelometer data were similar to those measured in other remote areas of the western USA. Quasi-dry chemical light scattering efficiencies derived using Mie theory were 3.6 m2 g(-1) for organic carbon, 2.5 m2 g(-1) for sulfates (ammonium sulfate and ammonium bisulfate), 2.6 m2 g(-1) for ammonium nitrate, and 1.76 m2 g(-1) for geological material. These values are lower than but consistent with previously reported results. Realistic efficiencies could not be derived using the multiple linear regression (MLR) approach. PMID- 11476488 TI - Phenanthrene degradation in soils co-inoculated with phenanthrene-degrading and biosurfactant-producing bacteria. AB - Contaminant sorption within the soil matrix frequently limits biodegradation. However, contaminant bioavailability can be species-specific. This study investigated bioavailability of phenanthrene (PHE) to two PHE-degrading bacteria (Pseudomonas strain R and isolate P5-2) in the presence of rhamnolipid biosurfactant and/or a biosurfactant-producing bacterium, Pseudomonas aeruginosa ATCC 9027. Pseudomonas strain R mineralized more soil-sorbed PHE than strain P5 2, but in aqueous cultures the rate and extent of PHE mineralization by P5-2 exceeded that by P. strain R. In Fallsington sandy loam (fine-loamy, mixed, active, mesic Typic Endoaquult) (high PHE-sorption capacity) the addition of rhamnolipid increased PHE mineralization by P. strain R. Phenanthrene mineralization in soils inoculated with P5-2 was minimal and no enhancement in PHE degradation was observed when biosurfactant was added. Co-inoculation of Fallsington sandy loam with the biosurfactant producer did not affect PHE mineralization by isolate P5-2, but significantly enhanced PHE mineralization by P. strain R. The enhancement of PHE mineralization could not be explained by P. aeruginosa-mediated PHE degradation. The addition of rhamnolipid at concentrations above the critical micelle concentration (CMC) resulted in enhanced PHE release from test soils. These results suggest that the PHE degrading strains were able to access different pools of PHE and that the biosurfactant-enhanced release of PHE from soils did not result in enhanced biodegradation. The results also demonstrated that bacteria with the catabolic potential to degrade sorbed hydrophobic contaminants could interact commensally with surfactant-producing strains by an unknown mechanism to hasten the biodegradation of aromatic hydrocarbons. Thus, understanding interactions among microbes may provide opportunities to further enhance biodegradation of soil bound organic contaminants. PMID- 11476489 TI - In situ bioremediation through mulching of soil polluted by a copper-nickel smelter. AB - Bioremediation of a heavy metal-polluted soil was investigated in a 3-yr field experiment by adding mulch to a polluted forest floor. The mulch consisted of a mixture of compost and woodchips. The remediation treatment decreased the toxicity of the soil solution to bacteria as determined by the [3H]-thymidine incorporation technique, that is, by measuring the growth rate of soil bacteria extracted from unpolluted humus after exposing them to soil solution containing heavy metals from the experimental plots. Canonical correlation analysis was performed in order to identify the chemical and microbiological changes in the soil. The pH of the mulched organic layer increased by one unit. The concentration of complexed Cu increased and that of free Cu2+ decreased in the soil solution from the mulch treatment. According to basal respiration and litter decomposition, microbial activity increased during the 3 yr following the remediation treatment. The [3H]-thymidine incorporation technique was also used to study the growth rate and tolerance of bacteria to Cu. The bacterial growth rate increased and the Cu tolerance decreased on the treated plots. The structure of the microbial community, as determined by phospholipid fatty acid (PLFA) analysis, remained unchanged. The results indicate that remediation of the polluted soil had occurred, and that adding a mulch to the forest floor is a suitable method for remediating heavy metal-polluted soil. PMID- 11476490 TI - Factors affecting the ratio of cation exchange capacity to clay content in lignite overburden. AB - Unusually high cation exchange capacity (CEC) values relative to clay content are frequently reported for lignite overburden and minesoils. The CEC to percent clay ratio is commonly greater than one and would require that the average charge of the clay fraction be greater than 100 cmol(c) kg(-1). A comparison of methods for particle-size distribution suggests that the major reason lignite overburden samples have CEC to percent clay ratios greater than one is incomplete dispersion of aggregates of clay minerals or shale fragments. Preliminary investigations revealed the presence of shale fragments, smectite, and partially weathered mica in the silt fraction. Methods commonly used in soil textural analysis underestimated clay content by approximately 24%. The silt fraction may, therefore, provide a "hidden" source of CEC. Another important factor influencing the CEC to percent clay ratio was the presence of organic materials (lignite) in the samples. Lignite may make a significant contribution to CEC in overburden materials. In a study designed to estimate the pH-dependent charge of both the mineral and organic fractions, the CEC of overburden organic constituents was determined to be approximately 158 cmol(c) kg(-1) at pH 8.2. The high CEC to percent clay ratio in lignite overburden and minesoils may be resolved by adjusting methods for clay determination to optimize dispersion and by accounting for CEC due to organic materials. An alternative approach is to use existing methodology and use correction factors to account for incomplete dispersion of clay minerals and the charge contributions of organic materials. PMID- 11476491 TI - Analysis of irrigation systems using sustainability-related criteria. AB - Sustainable agricultural development as a desired goal in irrigation management is a result of recent public awareness of the scarcity of water for food production. In order to incorporate sustainability-related criteria in the analysis of irrigation systems, the present study aims at introducing environmental indices that represent irrigation water conservation and satisfactory production and income for farmers under stress conditions. An experiment was conducted in Chania, Greece, during the irrigation periods of 1989 and 1990. The irrigation water delivered to 40 experimental plots and the relevant soil moisture content at the root zone were recorded. The data, collected in real time, were used for the calculation of the corresponding environmental indices. The variation of indices in time and space was high, and demonstrated that up to 13% of water was delivered to crops, 82% was yield loss, and 84% was economic return. The study indicated that environmental indices could be easily computed by means of routinely collected data, and could also be incorporated into decision-making approaches, such as compromise programming, in order to develop policies for irrigation water allocation. PMID- 11476492 TI - Revegetation of an abandoned uranium millsite on the Colorado Plateau, Arizona. AB - We attempted to restore native plants on disturbed sites at a former uranium mill on the Colorado Plateau near Tuba City, AZ. Four-wing saltbush [Atriplex canescens (Pursh) Nutt.] was successfully established in compacted caliche soil and in unconsolidated dune soil when transplants were irrigated through the first summer with 20 L/plant/wk. The caliche soil was ripped before planting to improve water-holding capacity. The diploid saltbush variety, angustifolia, had higher survival and growth than the common tetraploid variety, occidentalis, especially on dune soil. The angustifolia variety grew to 0.3 to 0.4 m3 per plant over 3 yr even though irrigation was provided only during the establishment year. By contrast, direct seeding of a variety of native forbs, grasses, and shrubs yielded poor results, despite supplemental irrigation throughout the first summer. In this arid environment (precipitation = 100 to 200 mm/yr), the most effective revegetation strategy is to establish keystone native shrubs, such as four-wing saltbush, using transplants and irrigation during the establishment year, rather than attempting to establish a diverse plant community all at once. PMID- 11476493 TI - Tracing nitrate transport and environmental impact from intensive swine farming using delta nitrogen-15. AB - Natural-abundance delta15N showed that nitrate generated from commercial land application of swine (Sus scrofa domesticus) waste within a North Carolina Coastal Plain catchment was being discharged to surface waters by ground water passing beneath the sprayfields and adjacent riparian buffers. This was significant because intensive swine farms in North Carolina are considered non discharge operations, and riparian buffers with minimum widths of 7.6 m (25 ft) are the primary regulatory control on ground water export of nitrate from these operations. This study shows that such buffers are not always adequate to prevent discharge of concentrated nitrate in ground water from commercial swine farms in the Mid-Atlantic Coastal Plain, and that additional measures are required to ensure non-discharge conditions. The median delta15N-total N of liquids in site swine waste lagoons was +15.4 +/- 0.2% vs. atmospheric nitrogen. The median delta15N-NO3 values of shallow ground water beneath and adjacent to site sprayfields, a stream draining sprayfields, and waters up to 1.5 km downstream were + 15.3 +/- 0.2 to + 15.4 +/- 0.2%. Seasonal and spatial isotopic variations in lagoons and well waters were greatly homogenized during ground water transport and discharge to streams. Neither denitrification nor losses of ammonia during spraying significantly altered the bulk ground water delta15N signal being delivered to streams. The lagoons were sources of chloride and potassium enrichment, and shallow ground water showed strong correlation between nitrate N, potassium, and chloride. The 15N-enriched nitrate in ground water beneath swine waste sprayfields can thus be successfully traced during transport and discharge into nearby surface waters. PMID- 11476494 TI - Nitrate and chloride loading to groundwater from an irrigated north-central U.S. sand-plain vegatable field. AB - Groundwater pollution and associated effects on drinking water have increased with the expansion of irrigated agriculture in north-central U.S. sand plains. Controlling this pollution requires an ability to measure and predict pollutant loading by specific agricultural systems. We measured NO3 and Cl loading to groundwater beneath a Wisconsin central sand plain irrigated vegetable field using both a budget method and a new monitoring-based method. By relying on frequent monitoring of shallow groundwater, the new method overcomes some limitations of other methods. Monitoring-based and budget methods agreed well, and indicated that loading to groundwater was 165 kg ha(-1) NO3-N and 111 kg ha( 1) Cl for sweet corn (Zea mays L.) in 1992, and 228 kg ha(-1) NO3-N and 366 kg ha(-1) Cl for potato (Solanum tuberosum L.) in 1993. Nitrate N loading was 56 to 60% of available N, or 66 to 70% of fertilizer N. Sweet corn NO3 loading was about typical for this region, but potato NO3 loading was probably 50% greater than typical because heavy rains provoked extra fertilizer application. Our results imply that typical NO3-N loading would be 119 kg ha(-1) for sweet corn and 203 kg ha(-1) for potato, even with strict adherence to University Extension fertilizer recommendations. To keep average groundwater NO3-N within the 10 mg L( 1) U.S. drinking water standard, each irrigated vegetable field would need to be offset by five to eight times as much land supplying NO3-free groundwater recharge. PMID- 11476495 TI - Controlling nitrate leaching in irrigated agriculture. AB - The impact of improved irrigation and nutrient practices on ground water quality was assessed at the Nebraska Management System Evaluation Area using ground water quality data collected from 16 depths at 31 strategically located multilevel samplers three times annually from 1991 to 1996. The site was sectioned into four 13.4-ha management fields: (i) a conventional furrow-irrigated corn (Zea mays L.) field; (ii) a surge-irrigated corn field, which received 60% less water and 31% less N fertilizer than the conventional field; (iii) a center pivot-irrigated corn field, which received 66% less water and 37% less N fertilizer than the conventional field; and (iv) a center pivot-irrigated alfalfa (Medicago sativa L.) field. Dating (3H/3He) indicated that the uppermost ground water was <1 to 2 yr old and that the aquifer water was stratified with the deepest water approximately 20 yr old. Recharge during the wet growing season in 1993 reduced the average NO3-N concentration in the top 3 m 20 mg L(-1), effectively diluting and replacing the NO3-contaminated water. Nitrate concentrations in the shallow zone of the aquifer increased with depth to water. Beneath the conventional and surge-irrigated fields, shallow ground water concentrations returned to the initial 30 mg NO3-N L(-1) level by fall 1995; however, beneath the center pivot irrigated corn field, concentrations remained at approximately 13 mg NO3-N L(-1) until fall 1996. A combination of sprinkler irrigation and N fertigation significantly reduced N leaching with only minor reductions (6%) in crop yield. PMID- 11476496 TI - A novel continuous-flow sequential extraction procedure for metal speciation in solids. AB - A continuous-flow extraction system was developed to speed up, facilitate, and improve the accuracy of the chemical fractionation of metals in solid materials. A three-step sequential extraction scheme was used to evaluate the novel system by analyzing calcium (Ca), iron (Fe), manganese (Mn), copper (Cu), and zinc (Zn) in a soil certified reference material (National Institute of Standards and Technology [NIST] SRM 2710). In the proposed system, extraction occurred in a closed chamber through which extractants were passed sequentially. The extracts were collected in a number of subfractions for subsequent name atomic absorption analysis. Apart from the advantages of simplicity, speed, and less risk of the contamination that flow analysis systems usually possess, the continuous-flow system can improve the accuracy of chemical fractionation of metals by sequential extraction. The system ensures that extraction is performed at designated pH values without any need of adjustment. Variation of sample weight to chamber volume ratios from 1:12 to 1:40 had no effect on the extractability of the metals studied. In the extraction of the acid soluble fraction, concentrations of acetic acid in the range 0.11 to 0.5 mol L(-1) had no significant effect on the amounts of metals extracted, except Fe. Increasing the concentration of hydroxylamine in the reducible fraction step from 0.04 to 0.5 mol L(-1) affected the extraction efficiency for Fe, Mn, and Zn. The extraction profile, rather than a single value of extracted concentration, of each element offers additional information about the kinetics of leaching processes and chemical associations between elements in the solid materials. PMID- 11476497 TI - Immobilization of cesium-137 and uranium in contaminated sediments using soil amendments. AB - Batch and dynamic leaching methods were used to evaluate the effectiveness of hydroxyapatite (HA), illite, and zeolite, alone and in combination, as soil additives for reducing the migration of cesium-137 (137Cs+) and uranium (U) from contaminated sediments. Amendment treatments ranging from 0 to 50 g kg(-1) were added to the sediment and equilibrated in 0.001 M CaCl2. After equilibration, the treatment supernatants were analyzed for 137Cs+, U, PO4, and other metals. The residual sediments were then extracted overnight using one of the following: 1.0 M NH4Cl, 0.5 M CaCl2, or the Toxicity Characteristic Leaching Procedure (TCLP) extractant. Cesium was strongly sorbed to the contaminated sediments, presumably due to interlayer fixation within native illitic clays. In fact, 137Cs+ was below detection limits in the initial equilibration solutions, the CaCl2 extract, and the TCLP solution, regardless of amendment. Extractants selective for interlayer cations (1.0 M NH4Cl) were necessary to extract measurable levels of 137Cs+. Addition of illitic clays further reduced Cs+ extractability, even when subjected to the aggressive extractants. Zeolite, however, was ineffective in reducing Cs+ mobility when subjected to the aggressive extractants. Hydroxyapatite was less effective than illite at reducing NH4+-extractable Cs+. Hydroxyapatite, and mixtures of HA with illite or zeolite, were highly effective in reducing U extractability in both batch and leaching tests. Uranium immobilization by HA was rapid with similar final U concentrations observed for equilibration times ranging from 1 h to 30 d. The current results demonstrate the effectiveness of soil amendments in reducing the mobility of U and Cs+, which makes in-place immobilization an effective remediation alternative. PMID- 11476498 TI - In situ stabilization of soil lead using phosphorus. AB - In situ stabilization of Pb-contaminated soils can be accomplished by adding phosphorus. The standard remediation procedure of soil removal and replacement currently used in residential areas is costly and disruptive. This study was carried out to evaluate the influence of P and other soil amendments on five metal-contaminated soils and mine wastes. Seven treatments were used: unamended control; 2,500 mg of P/kg as triple superphosphate (TSP), phosphate rock (PR), acetic acid followed by TSP, and phosphoric acid (PA); and 5,000 mg of P/kg as TSP or PR. A significant reduction in bioavailable Pb, as determined by the physiologically based extraction test (PBET), compared with the control upon addition of P was observed in all materials tested. Increasing the amount of P added from 2,500 to 5,000 mg/kg also resulted in a significantly greater reduction in bioavailable Pb. Phosphate rock was equally or more effective than TSP or PA in reducing bioavailable Pb in four out of five soils tested. Preacidification produced significantly lower bioavailable Pb compared with the same amount of P from TSP or PR in only one material. Reductions in Pb bioavailability as measured by PBET were evident 3 d after treatment, and it may indicate that the reactions between soil Pb and P occurred in situ or during the PBET. No further reductions were noted over 365 d. X-ray diffraction data suggested the formation of pyromorphite-like minerals induced by P additions. This study suggests that P addition reduced bioavailable Pb by PBET and has potential for in situ remediation of Pb-contaminated soils. PMID- 11476499 TI - Chemical immobilization of lead, zinc, and cadmium in smelter-contaminated soils using biosolids and rock phosphate. AB - Chemical immobilization, an in situ remediation method where inexpensive chemicals are used to reduce contaminant solubility in contaminated soil, has gained attention. We investigated the effectiveness of lime-stabilized biosolid (LSB), N-Viro Soil (NV), rock phosphate (RP), and anaerobic biosolid (AB) to reduce extractability and plant and gastrointestinal (GI) bioavailability in three Cd-, Pb-, and Zn-contaminated soils from smelter sites. Treated (100 g kg( 1) soil) and control soils were incubated at 27 degrees C and -0.033 MPa (0.33 bar) water content for 90 d. The effect of soil treatment on metal extractability was evaluated by sequential extraction, on phytoavailability by a lettuce bioassay (Lactuca sativa L.), on human GI availability of Pb from soil ingestion by the Physiologically Based Extraction Test. The largest reductions in metal extractability and phytoavailability were from alkaline organic treatments (LSB and NV). Phytotoxic Zn [1188 mg Zn kg(-1) extracted with 0.5 M Ca(NO3)2] in Blackwell soil (disturbed soil) was reduced by LSB, NV, and RP to 166, 25, and 784 mg Zn kg(-1), respectively. Rock phosphate was the only treatment that reduced GI-available Pb in both gastric and intestinal solutions, 23 and 92%, respectively. Alkaline organic treatments (LSB, NV) decreases Cd transmission through the food chain pathway, whereas rock phosphate decreases risk from exposure to Pb via the soil ingestion pathway. Alkaline organic treatments can reduce human exposure to Cd and Pb by reducing Zn phytotoxicity and revegetation of contaminated sites. PMID- 11476500 TI - Assessment of metal availability in smelter soil using earthworms and chemical extractions. AB - Chemical immobilization is a relatively inexpensive in situ remediation method that reduces soil contaminant solubility, but the ability of this remediation treatment to reduce heavy metal bioavailability and ecotoxicity to soil invertebrates has not been evaluated. Our objectives were to (i) assess the ability of chemical immobilization amendments (municipal sewage sludge biosolids and rock phosphate) to reduce metal bioavailability and toxicity in a toxic metal contaminated smelter soil and (ii) evaluate soil extraction methods using Ca(NO3)2 solution or ion-exchange membranes coated with diethylenetriaminepentaacetic acid (DTPA) as surrogate measures of metal bioavailability and ecotoxicity. We treated a soil contaminated by Zn and Pb milling and smelting operations and an uncontaminated control soil with lime stabilized municipal biosolids (LSB), rock phosphate (RP), or anaerobically digested municipal biosolids (SS) and evaluated lethality of the remediated soils to earthworm (Eisenia fetida Savigny). Lime-stabilized municipal biosolids was the only remediation amendment to successfully immobilize lethal levels of Zn in the smelter soil (14-d cumulative mortality < or = 15%). Calcium nitrate extractable Zn in the lethal Zn smelter soil-amendment combinations was 11.5 to 18.2 mmol/kg, compared with the nonlethal LSB amended soil (0.62 mmol/kg). The Ca(NO3)2-extractable Zn-based median lethal concentration (LC50) of 6.33 mmol/kg previously developed in Zn-spiked artificial soils was applicable in the remediated smelter soils despite a 14-fold difference in total Zn concentration. Chelating ion-exchange membrane uptake among the soils was highly variable (mean CV = 39%) compared with the Ca(NO3)2-extraction (mean CV = 1.9%) and not well related to earthworm toxicity. PMID- 11476501 TI - Sources of salinity near a coal mine spoil pile, north-central Colorado. AB - A small (1 km2) salt-affected stream drainage on the High Plains north of Denver, Colorado was sampled to determine the near-surface dispersion of soluble salts and metals from low-sulfur coal mining waste (spoil). Surface waters collected along the 0.8-km stream reach, and aqueous leachates of spoil and naturally saline local soil, were analyzed for chemical constituents and sulfur isotopes. In this semiarid setting with abundant carbonate-bearing surficial sediments, the limited, mildly acidic drainage from the spoil pile is quickly neutralized, restricting the mobility of many elements. However, some spoil-derived constituents were clearly traceable within the upper 0.4 km of the stream reach. Spoil leachates and surface water near the spoil pile have distinctive compositions of major anions and cations, and elevated levels of dissolved nitrate compared with downstream waters. Spoil-derived sulfate was traceable because it has generally positive values of delta34S that contrasted with generally negative values of delta34S in soil leachates and evaporite salts from the surrounding area. Spatial-chemical sampling of surface water showed an abrupt increase in dissolved U, Se, B, Li, and Mn in the lower 0.4 km of the stream reach where shallow ground water from surrounding irrigated fields contributed to surface flow. The downstream evolution of surface water chemistry and sulfur isotopic composition is consistent with mixing between spoil-affected upstream water and irrigation-return water. The methods described should be applicable at other sites in similar settings where the environmental effect of low-sulfur coal mining waste must be assessed and where access to samples of shallow ground water is limited. PMID- 11476502 TI - Factors controlling sediment and phosphorus export from two Belgian agricultural catchments. AB - Sediment and total phosphorus (TP) export vary through space and time. This study was conducted to determine the factors controlling sediment and TP export in two agricultural catchments situated in the Belgian Loess Belt. At the outlet of these catchments runoff discharge was continuously measured and suspended sediment samples were taken during rainfall events. Within the catchments vegetation type and cover, soil surface parameters, erosion features, sediment pathways, and rainfall characteristics were monitored. Total P content and sediment characteristics such as clay, organic carbon, and suspended sediment concentration were correlated. Total sediment and TP export differ significantly between the monitored catchments. Much of the difference is due to the occurrence of an extreme event in one catchment and the morphology and spatial organization of land use in the catchments. In one catchment, the direct connection between erosive areas and the catchment outlet by means of a road system contributed to a high sediment delivery ratio (SDR) at the outlet. In the other catchment, the presence of a wide valley in the center of the catchment caused sediment deposition. Vegetation also had an effect on sediment production and deposition. Thus, many factors control sediment and TP export from small agricultural catchments; some of these factors are related to the physical catchment characteristics such as morphology and landscape structure and are (semi)permanent, while others, such as vegetation cover and land use, are time dependent. PMID- 11476503 TI - Effects of dissolved organic matter from animal waste effluent on chlorpyrifos sorption by soils. AB - The increased use of animal waste-derived effluents for irrigation could result in the enhanced movement of pesticides through complexation with dissolved organic materials. Batch equilibrium studies were conducted to measure the interaction among soil, chlorpyrifos [O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate], and dissolved organic matter (DOM) from poultry, swine, and cow waste-derived lagoon effluents. All DOM was found to have a strong affinity for chlorpyrifos, resulting in reduced sorption of chlorpyrifos by soil, thus the potential for DOM-enhanced mobility. Effluent DOM was observed to sorb to soils. Thus, for increasingly higher soil mass to solution volume ratios, the effect of chlorpyrifos association with water-borne DOM on sorption decreases significantly. For high soil mass to solution volume ratios typical of soil profiles in the landscape, the potential for DOM-enhanced transport will be greatly attenuated. Dissolved organic matter concentration and the nonpolar nature of DOM in the lagoon effluent decreased with increasing residence time in the cells of the lagoon system, thus reducing the potential for DOM-enhanced transport. PMID- 11476504 TI - Influence of plant growth on degradation of linear alkylbenzene sulfonate in sludge-amended soil. AB - Widespread application of sewage sludge to agricultural soils in Denmark has led to concern about the possible accumulation and effects of linear alkylbenzene sulfonate (LAS) in the soil ecosystem. Therefore, we have studied the uptake and degradation of LAS in greenhouse pot experiments. Sewage sludge was incorporated into a sandy soil to give a range from very low to very high applications (0.4 to 90 Mg dry wt. ha(-1)). In addition, LAS was added as water solutions. The soil was transferred to pots and sown with barley (Hordeum vulgare L. cv. Apex), rape (Brassica napus L. cv. Hyola 401), or carrot (Daucus carota L.). Also, plant-free controls were established. For all additions there was no plant uptake above the detection limit at 0.5 mg LAS kg(-1) d.w, but plant growth stimulated the degradation. With a growth period of 30 d, LAS concentrations in soil from pots with rape had dropped from 27 to 1.4 mg kg(-1) dry wt., but in plant-free pots the concentration decreased only to 2.4 mg kg(-1) dry wt. When LAS was added as a spike, the final concentration in soil from planted pots was 0.7 mg kg(-1) dry wt., but in pots without plants the final concentration was much higher (2.5 mg kg(-1) dry wt.). During degradation, the relative fraction of homologues C10, C11, and C12 decreased, while C13 increased. PMID- 11476505 TI - Anaerobic degradation of atrazine and metolachlor and metabolite formation in wetland soil and water microcosms. AB - The half-lives, degradation rates, and metabolite formation patterns of atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine) and metolachlor [2 chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamide] were determined in an anaerobic wetland soil incubated at 24 degrees C for 112 d. At 0, 7, 14, 28, 42, 56, and 112 d, the soil and water were analyzed for atrazine and metolachlor, and their major metabolites. The soil oxidation-reduction potential reached -200 mV after 14 d. Degradation reaction rates were first-order for atrazine in anaerobic soil and for metolachlor in the aqueous phase. Zero order reaction rates were best fit for atrazine in the aqueous phase and metolachlor in anaerobic soil. In anaerobic soil, the half-life was 38 d for atrazine and 62 d for metolachlor. In the aqueous phase above the soil, the half life was 86 d for atrazine and 40 d for metolachlor. Metabolites detected in the anaerobic soil were hydroxyatrazine and deethylatrazine for atrazine, and relatively small amounts of ethanesulfonic acid and oxanilic acid for metolachlor. Metabolites detected in the aqueous phase above the soil were hydroxyatrazine, deethylatrazine, and deisopropylatrazine for atrazine, and ethanesulfonic acid and oxanilic acid for metolachlor. Concentrations of metabolites in the aqueous phase generally peaked within the first 25 d and then declined. Results indicate that atrazine and metolachlor can degrade under strongly reducing conditions found in wetland soils. Metolachlor metabolites, ethanesulfonic acid, and oxanilic acid are not significantly formed under anaerobic conditions. PMID- 11476506 TI - Degradation of soil fumigants as affected by initial concentration and temperature. AB - Soil fumigation using shank injection creates high fumigant concentration gradients in soil from the injection point to the soil surface. A temperature gradient also exists along the soil profile. We studied the degradation of methyl isothiocyanate (MITC) and 1,3-dichloropropene (1,3-D) in an Arlington sandy loam (coarse-loamy, mixed, thermic Haplic Durixeralf) at four temperatures and four initial concentrations. We then tested the applicability of first-order, half order, and second-order kinetics, and the Michaelis-Menten model for describing fumigant degradation as affected by temperature and initial concentration. Overall, none of the models adequately described the degradation of MITC and 1,3 D isomers over the range of the initial concentrations. First-order and half order kinetics adequately described the degradation of MITC and 1,3-D isomers at each initial concentration, with the correlation coefficients greater than 0.78 (r2> 0.78). However, the derived rate constant was dependent on the initial concentration. The first-order rate constants varied between 6 and 10x for MITC for the concentration range of 3 to 140 mg kg(-1), and between 1.5 and 4x for 1,3 D isomers for the concentration range of 0.6 to 60 mg kg(-1), depending on temperature. For the same initial concentration range, the variation in the half order rate constants was between 1.4 and 1.7x for MITC and between 3.1 and 6.1x for 1,3-D isomers, depending on temperature. Second-order kinetics and the Michaelis-Menten model did not satisfactorily describe the degradation at all initial concentrations. The degradation of MITC and 1,3-D was primarily biodegradation, which was affected by temperature between 20 and 40 degrees C, following the Arrhenius equation (r2 > 0.74). PMID- 11476507 TI - Organochlorine pollutants in remote mountain lake waters. AB - Hexachlorocyclohexanes (HCHs; alpha- and gamma-isomers), endosulfans (alpha- and beta-isomers and the sulfate residue), hexachlorobenzene (HCB), dichlorodiphenyltrichloroethane (DDTs), and polychlorobiphenyls (PCBs) were measured in waters from three European remote mountain lakes situated in the Alps, Pyrenees, and Caledonian mountains. Sampling encompassed both ice-free and ice-covered periods at different water column depths. High HCH concentrations were found in all lakes, those in the Alps and Pyrenees (990-2,900 pg/L) being among the highest recorded in continental waters. Endosulfans and endosulfan sulfate (120-1,150 pg/L) were the second major group of organochlorine contaminants, showing a remarkable stability upon atmospheric long-range transport. The concentrations of HCB, DDTs, and PCB (4-8, 0.6-16, and 26-110 pg/L, respectively) were low in comparison with other continental waters. Hexachlorocyclohexanes, endosulfans, and HCB were essentially found in the dissolved phase. Phase partitioning of the more hydrophobic compounds exhibited a dependence on temperature and water-suspended particles. Comparison between different sampling seasons and water depths indicated a remarkable concentration uniformity within lake, but major interlake differences. Normalization to turnover rates showed higher interlake similarity. Preferential accumulation of the less volatile compounds in the Alp lake and significant increase of baseline contributions of organochlorine compounds and residues in the Caledonian lake are also evidenced from these turnover rates. PMID- 11476508 TI - Forest fertilization with wood ash: effect on the distribution and storage of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs). AB - Before wood ash can be safely used as a fertilizer in forests, possible negative effects such as input of organic contaminants or remobilization of contaminants already stored in the soil must be investigated. The objective of this study was to examine the effects of wood ash application on concentrations, storage, and distribution of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) in a Swiss forest soil. In May 1998, we added 8 Mg wood ash ha( 1) to a forest soil. We determined 20 PAHs and 14 PCBs in the organic layer, in the bulk mineral soil, and in soil material taken from preferential flow paths and from the matrix before and after the wood ash application. In the control plots, the concentrations of PAHs in the organic layer indicated moderate pollution (sum of 20 PAHs: 0.8-1.6 mg kg(-1)), but sum of PCB concentrations was high (21-48 microLg kg(-1)). The wood ash had high concentrations of PAHs (sum of 20 PAHs: 16.8 mg kg(-1)), but low concentrations of PCBs (sum of 14 PCBs: 3.4 microg kg(-1)). The wood ash application increased the PAH concentrations in the organic horizons up to sixfold. In contrast, PCB concentrations did not change in the Oa horizon and decreased up to one third in the Oi and Oe horizons. The decrease was probably caused by the mobilization of stored PCBs because of the high pH of the wood ash. This probably results in a higher mobility of dissolved organic matter, acting as PCB carrier. In the mineral soil, the preferential flow paths of the A horizon contained more PAHs and PCBs (+20 +/- 15% and +43 +/- 60%, respectively) than the matrix. This was particularly true for higher molecular weight compounds (molecular weight > 200 g mol(-1)). Below 50 cm depth, concentrations of PAHs and PCBs were smaller in the preferential flow paths, suggesting that in deeper depths, processes acting as sinks dominated over inputs in the preferential flow paths. PMID- 11476509 TI - Nitrate loss in subsurface drainage as affected by nitrogen fertilizer rate. AB - The relationships between N fertilizer rate, yield, and NO3 leaching need to be quantified to develop soil and crop management practices that are economically and environmentally sustainable. From 1996 through 1999, we measured yield and NO3 loss from a subsurface drained field in central Iowa at three N fertilizer rates: a low (L) rate of 67 kg ha(-1) in 1996 and 57 kg ha(-1) in 1998, a medium (M) rate of 135 kg ha(-1) in 1996 and 114 kg ha(-1) in 1998, and a high (H) rate of 202 kg ha(-1) in 1996 and 172 kg ha(-1) in 1998. Corn (Zea mays L.) and soybean [Glycine max (L.) Merr.] were grown in rotation with N fertilizer applied in the spring to corn only. For the L treatment, NO3 concentrations in the drainage water exceeded the 10 mg N L(-1) maximum contaminant level (MCL) established by the USEPA for drinking water only during the years that corn was grown. For the M and H treatments, NO3 concentrations exceeded the MCL in all years, regardless of crop grown. For all years, the NO3 mass loss in tile drainage water from the H treatment (48 kg N ha(-1)) was significantly greater than the mass losses from the M (35 kg N ha(-1)) and L (29 kg N ha(-1)) treatments, which were not significantly different. The economically optimum N fertilizer rate for corn was between 67 and 135 kg ha(-1) in 1996 and 114 and 172 kg ha(-1) in 1998, but the net N mass balance indicated that N was being mined from the soil at these N fertilizer levels and that the system would not be sustainable. PMID- 11476510 TI - Phosphorus runoff from two water sources on a calcareous soil. AB - Phosphorus (P) in irrigation runoff may enrich offsite water bodies and streams and be influenced by irrigation water quality and antecedent soil surface conditions. Runoff, soil loss, and P fractions in runoff using reverse osmosis (RO) water or mixed RO and well water (RO/ Tap) were studied in a laboratory sprinkler study to evaluate water source effects on P transport. A top- or subsoil Portneuf silt loam (coarse-silty, mixed, superactive, mesic Durinodic Xeric Haplocalcid), either amended or not amended with manure and/or with cheese whey, with Olsen P from 20 to 141 mg kg(-1) and lime from 108 to 243 g kg(-1), was placed in 1.5 x 1.2 x 0.2-m-deep containers with 2.4% slope and irrigated three times from a 3-m height for 15 min, applying 20 mm of water. The first irrigation was on a dry loose surface, the second on a wet surface, and the third on a dry crusted surface. Surface (ca. 2 cm) soil samples, prior to the first irrigation, were analyzed for Olsen P, water-soluble P (Pws), and iron-oxide impregnated paper-extractable P (FeO-P) analyses. Following each irrigation we determined runoff, sediment, dissolved reactive phosphorus (DRP) in a 0.45-microm filtered sample, and FeO-P and total P in unfiltered samples. Soil surface conditions had no effect on P runoff relationships. Water source had no significant effect on the relationship between DRP or FeO-P runoff and soil test P, except for DRP in RO runoff versus water-soluble soil P (r2 = 0.90). Total P in RO runoff versus soil P were not related; but weakly correlated for RO/Tap (r2 < 0.50). Water source and soil surface conditions had little or no effect on P runoff from this calcareous soil. PMID- 11476511 TI - Effects of tillage and phosphorus placement on phosphorus runoff losses in a grain sorghum-soybean rotation. AB - Phosphorus enhances eutrophication of fresh water bodies. This study was conducted to determine the influence of tillage and P placement on P losses in runoff water from a somewhat poorly drained soil (Woodson silt loam [fine, smectitic, thermic Abruptic Argiaquoll], 1.0-1.5% slope) in a grain sorghum [Sorghum bicolor (L.) Moenchl-soybean [Glycine mar (L.) Merr] rotation. Chisel disk-field cultivate (ChT), ridge-till (RT), and no-till (NT) in combination with 0 kg P ha(-1) or 24 kg P ha(-1) broadcast or knifed (applied prior to planting grain sorghum) were studied. Runoff volume and losses of sediment and P were summed over the growing season. Significant interactions between tillage and P placement for soluble P losses were found. For example, soluble P loss in 1999 for NT-broadcast in grain sorghum was 358 g ha(-1); significantly greater than 31 g ha(-1) for NT-knife or 23 g ha(-1) for NT-check. Similar results were found for RT but no such differences were found for ChT. Bioavailable P losses were generally highest with broadcast P placement and for NT and RT. Total P losses were significantly higher at 959 g ha(-1) with broadcast P on grain sorghum in 1998, compared with 521 g ha(-1) for the check and 659 g ha(-1) for the knifed P applications. Total P losses in 1999 for soybeans were only 18 g ha(-1) for NT, which was significantly lower than 75 g ha(-1) for ChT and 66 g ha(-1) for RT. The results indicate that broadcast P applications on RT and NT will increase P losses, but the influence of tillage was not consistent. PMID- 11476512 TI - Controlled drainage and wetlands to reduce agricultural pollution: a lysimetric study. AB - Controlled drainage and wetlands could be very effective practices to control nitrogen pollution in the low-lying agricultural plains of northeast Italy, but they are not as popular as in other countries. An experiment on lysimeters was therefore carried out in 1996-1998, with the double aim of obtaining local information to encourage the implementation of these practices and to gain more knowledge on the effects involved. Controlled drainage + subirrigation and wetlands were all considered as natural systems where alternative water table management could ameliorate water quality, and were compared with a typical water management scheme for crops in the open field. Eight treatments were considered: free drainage on maize (Zea mays L.) and sugarbeet (Beta vulgaris L.), two treatments of controlled drainage on the same crops, and five wetland treatments using common reed [Phragmites australis (Cav.) Trin. ex Steud.], common cattail (Typha latifolia L.), and tufted sedge (Carex elata All.), with different water table or flooding levels. Lysimeters received about 130 g m 2 of N with fertilization and irrigation water, with small differences among treatments. The effects of treatments were more evident for NO3-N concentrations than for the other chemical parameters (total Kjeldahl nitrogen, pH, and electrical conductivity), with significantly different medians among free drainage (33 mg L( 1)), controlled drainage (1.6 and 2.6 mg L(-1)), and wetlands (0.5-0.7 mg L(-1)). Referring to free drainage, NO3-N losses were reduced by 46 to 63% in controlled drainage and 95% in the average of wetlands. Wetlands also reduced losses of total dissolved solids from 253 g m(-2) (average of crop treatments) to 175 g m( 2) (average of wetlands). PMID- 11476513 TI - Radionuclide transport above a near-surface water table: III. Soil migration and crop uptake of three gamma-emitting radionuclides, 1990 to 1993. AB - This paper summarizes the vertical distributions of 22Na, 137Cs, and 60Co above controlled water tables in deep and shallow lysimeters during a four-year experiment. The activity concentration profiles were all determined at the time of harvest of a winter wheat (Triticum aestivum L. cv. Pastiche) crop. Activity concentrations in different crop tissues were determined and crop uptake expressed as both an inventory ratio (IR) and a transfer factor (TFw), weighted to account for root and radionuclide distributions within the soil profile. Experimental variates were subjected to analysis of variance to determine the single and combined effects of the soil depth and the year of the experiment on the results obtained. Each radionuclide showed significant variations in activity concentration with soil depth, but the significance of these variations from year to year was dependent on radionuclide. A distinction in the behavior of weakly sorbed (22Na) and more highly sorbed (137Cs and 60Co) radionuclides was observed. The former exhibited significant variations in its distribution in the soil profile from year-to-year whereas the latter did not. Relatively high TF, values for 22Na were maintained throughout the experiment, whereas for 137Cs and 60Co, the highest TFw values were recorded in 1990 followed by a significant decline in 1991, with TFw remaining low in 1992 and 1993. The TFw values were, in general, significantly higher for deep lysimeters than for shallow lysimeters. This is thought to provide evidence of enhanced radionuclide absorption by the relatively small fraction of roots in the vicinity of the deeper water table. PMID- 11476514 TI - Inverse analyses of transport of chlorinated hydrocarbons subject to sequential transformation reactions. AB - Chemical and biological transformations can significantly affect contaminant transport in the subsurface. To better understand such transformation reactions, an equilibrium-nonequilibrium sorption transport model, HYDRUS-1D, was modified by including inverse solutions for multiple breakthrough curves resulting from the transport of solutes undergoing sequential transformations. The inverse solutions were applied to miscible-displacement experiments involving dissolved concentrations of trichloroethylene (TCE) undergoing reduction and/or transformations in the presence of zero-valent metal porous media (i.e., iron or copper-coated iron filings) to produce ethylene. The inverse model solutions provided a reasonable description of the transport and transformation processes. Simultaneous fitting of multiple breakthrough curves of TCE and ethylene placed additional constraints on the inverse solution and improved the reliability of parameter estimates. Confidence intervals of optimized parameters were reduced significantly in comparison with those obtained by fitting TCE breakthrough curves independently. Further evidence for accurate parameter estimates was given when the parameter values agreed with previously reported values from independent batch and degradation experiments. Optimized values of the normalized degradation rates for the equilibrium (1.4 x 10(-4) to 7.2 x 10(-5) L h(-1)m(-2)) and nonequilibrium (1.2 x 10(-4) to 5.5 x 10(-5)L h(-1)m(-2)) models compared well with values (0.03 to 6.5 x 10(-5) L h(-1) m(-2)) obtained from previous studies. The estimated TCE-iron sorption coefficients (0.52 to 2.85 L kg(-1)) were also consistent with a previously reported value (1.47 L kg(-1)). PMID- 11476515 TI - Seasonal leaching and biodegradation of dicamba in turfgrass. AB - The leaching of surface-applied herbicides, such as dicamba (2methoxy-3,6 dichlorobenzoic acid), to ground water is an environmental concern. Seasonal changes in soil temperature and water content, affecting infiltration and biodegradation, may control leaching. The objectives of this study were to (i) investigate the leaching of dicamba applied to turfgrass, (ii) measure the degradation rate of dicamba in soil and thatch in the laboratory under simulated field conditions, and (iii) test the ability of the model EXPRES (containing LEACHM) to simulate the field transport and degradation processes. Four field lysimeters, packed with sandy loam soil and topped with Kentucky bluegrass (Poa pratensis L.) sod, were monitored after receiving three applications (May, September, November) of dicamba. Concentrations of dicamba greater than 1 mg L( 1) were detected in soil water. Although drying of the soil during the summer prevented deep transport, greater leaching occurred in late autumn due to increased infiltration. From the batch experiment, the degradation rate for dicamba in thatch was 5.9 to 8.4 times greater than for soil, with a calculated half-life as low as 5.5 d. Computer modeling indicated that the soil and climatic conditions would influence the effectiveness of greater degradation in thatch for reducing dicamba leaching. In general, EXPRES predictions were similar to observed concentration profiles, though peak dicamba concentrations at the 10-cm depth tended to be higher than predicted in May and November. Differences between predictions and observations are probably a result of minor inaccuracies in the water-flow simulation and the model's inability to modify degradation rates with changing climatic conditions. PMID- 11476516 TI - Effect of flue gas desulfurization (FGD) by-product on water quality at an underground coal mine. AB - In this paper, a field study was carried out to examine the effect of flue gas desulfurization (FGD) by-product on water quality at an underground coal mine in central-eastern Ohio. Flue gas desulfurizalion by-product was injected into the down-dip portions of the Robert-Dawson mine in an attempt to seal major seeps exiting the mine and to coat exposed pyritic surfaces. Immediately following grout injection, significant increases in acidity, iron, aluminum, sulfur, and calcium were observed at most surface and ground water locations near where grouting was carried out. Following this initial flush of elements, concentrations of most constituents have decreased to near pre-grouting levels. Data from the site and geochemical modeling suggest that an increase in water level or rerouting of drainage flow resulted in the dissolution of iron and aluminum sulfate salts and ferrihydrite. Dissolution of the FGD grout material resulted in increases in calcium and sulfate concentrations in the drainage waters. Water within the mine voids was saturated with respect to calcium sulfate and gypsum immediately following grout injection. Based on an analysis of core samples obtained from the site, acid mine drainage (AMD) was in contact with at least some portions of the grout and this resulted in grout weathering. Subsequent transport of calcium and sulfate to the underclay, perhaps by fracture flow, has resulted in the deposition of gypsum and calcium sulfate solids. PMID- 11476517 TI - Evaluation of leachates from coal refuse blended with fly ash at different rates. AB - There is great interest in returning coal combustion products to mining sites for beneficial reuse as liming agents. A column study examined the effects of blending two coal fly ashes with an acid-forming coal refuse (4% pyritic S). Both fly ashes were net alkaline, but had relatively low neutralizing capacities. One ash with moderate alkalinity (CRF) was bulk blended with coal refuse at 0, 20, and 33% (w/w), while another lower alkalinity ash (WVF) was blended at 0, 5, 10, 20, and 33% (w/w). The columns were leached (unsaturated) weekly with 2.5 cm of simulated precipitation for >150 wk. Where high amounts of ash alkalinity (>20% w/w) were mixed with the coal refuse, pyrite oxidation was controlled and leachate pH was >7.0 with low metal levels throughout the study. At lower rates of alkalinity loading, trace metals were sequentially released from the WVF ash as the 5, 10, and 20% treatments acidified due to pyrite oxidation. Lechate metals increased in proportion to the total amounts applied in the ash. In this strongly acidic environment, metals such as Mn, Fe, and Cu were dissolved and leached from the ash matrix in large quantities. If ash is to be beneficially reused in the reclamation of acid-producing coal refuse, the alkalinity and potential acidity of the materials must be balanced through the appropriate addition of lime or other alkaline materials to the blend. Highly potentially acidic refuse material, such as that used here, may not be suitable for ash/refuse codisposal scenarios. PMID- 11476518 TI - Oxidation kinetics of the combustible fraction of construction and demolition wastes. AB - Proper disposal of construction and demolition wastes (CDW) has received wide attention recently due to significantly large quantities of waste streams collected from razed or retrofitted buildings in many metropolitan regions. Burning the combustible fractions of CDW (CCDW) and possibly recovering part of the heat content for economic uses could be valuable for energy conservation. This paper explores the oxidation kinetics of CCDW associated with its ash characterization. Kinetic parameters for the oxidation of CCDW were numerically calculated using thermal gravimetric analysis (TGA) and the resultant rate equations were therefore developed for illustrating the oxidation processes of CCDW simultaneously. Based on three designated heating rates, each of the oxidation processes can be featured distinctively with five different stages according to the rate of weight change at the temperature between 300 K and 923 K. In addition, Fourier transform infrared (FTIR) spectroscopy was employed, associated with a lab-scale fixed-bed incinerator for monitoring the composition of flue gas. Carbon dioxide (CO2) was found as a major component in the flue gas. The fuel analysis also included an ash composition analysis via the use of X-ray powder diffraction (XRD), atomic absorption (AA) spectroscopy, inductively coupled plasma-atomic emission spectroscopy (ICP-AES), and scanning electron microscopy-energy dispersive spectroscopy (SEM-EDX). The ash streams were identified as nonhazardous materials based on the toxicity characteristic leaching procedure (TCLP). Overall, the scientific findings gained in this study will be helpful for supporting a sound engineering design of real-world CCDW incineration systems. PMID- 11476519 TI - Decomposition and nitrogen mineralization from biosolids and other organic materials: relationship with initial chemistry. AB - Biosolids are effective forest fertilizers. In order to facilitate their use it is important that one be able to predict the amount and rate of mineralization of nutrients, particularly nitrogen, and the relationship between substrate chemistry and N release. We examined the relationships between substrate quality and nitrogen release in a variety of organic materials. Rates of decomposition and net N mineralization from four biosolids, wheat straw, paper fines, and Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] needle litter were measured during 391-d incubations in a greenhouse, and at two field sites in wet coastal and dry interior forests. Decomposition rates were best predicted by a model incorporating the ratio of carbon to organic matter. The decomposition model extrapolated well to the field when site-specific correction factors were applied. There was a weak relationship between rates of decomposition and net N mineralization. Rates of net N mineralization were best predicted by a model incorporating the initial organic N concentration and the proportion of phenolic C determined from solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. The mineralization model extrapolated less well to the field, but the effect of substrate chemistry was still apparent. Among the four biosolids there was a strong correlation between organic N concentration and indices or protein determined from 13C NMR, suggesting that these protein indices may be useful for predicting N mineralization from biosolids. There was some evidence that the protein content and N mineralization in biosolids may be predictable from the sewage treatment process employed. PMID- 11476520 TI - Denitrification at a long-term forested land treatment system in the Piedmont of Georgia. AB - Spray irrigation of forested land can provide an effective system for nutrient removal and treatment of municipal wastewater. Evolution of N2 + N2O from denitrifying activity is an important renovation pathway for N applied to forested land treatment systems. Federal and state guidance documents for design of forested land treatment systems indicate the expected range for denitrification to be up to 25% of applied N, and most forest land treatment systems are designed using values from 15 to 20% of applied N. However, few measurements of denitrification following long-term wastewater applications at forested land treatment sites exist. In this study, soil N2 + N2O-N evolution was directly measured at four different landscape positions (hilltop, midslope, toe slope, and riparian zone) in a forested land treatment facility in the Georgia Piedmont that has been operating for more than 13 yr. Denitrification rates within effluent-irrigated areas were significantly greater than rates in adjacent nonirrigated buffer zones. Rates of N2 + N2O-N evolved from soil in irrigated forests ranged from 5 to 10 kg ha(-1) yr(-1) N on the three upland landscape positions and averaged 38 kg ha(-1) yr(-1) N within the riparian zone. The relationship between measured riparian zone denitrification rates and soil physical and chemical properties was poor. The best relationship was with soil temperature, with an r2 of 0.18. Overall, on a landscape position weighted basis, only 2.4% of the wastewater-applied N was lost through denitrification. PMID- 11476521 TI - Acidification-neutralization processes in a lignite mine spoil amended with fly ash or limestone. AB - A laboratory experiment was conducted to investigate the longterm effects of amending sulfide-rich lignite mine spoil with fly ash (originating from a coal fired power station and largely comprised of aluminosilicates) and/or agricultural limestone. The experiment was carried out with soil moisture maintained at field capacity or alternate cycles of wetting and drying. Results obtained suggest that the principal acidification processes were oxidation of sulfide and formation of hydroxysulfate (FeOHSO4), whereas the main neutralization processes were weathering of aluminosilicates in fly ash-treated samples and dissolution of calcium carbonate in limestone-treated samples. The highest dose of limestone rapidly raised the pH of the spoil, but this increase was not maintained throughout the one-year experiment. In contrast, fly ash treated samples showed a more sustained increase in pH, attributable to the gradual weathering of aluminosilicates. The best results (i.e., good short- and long-term neutralization) were obtained in samples treated with both fly ash and limestone. The low liming capacity of the fly ash (47.85 cmol kg(-1)) means that it must be used in large quantities, an advantage in achieving the further aim of disposing of the fly ash. PMID- 11476522 TI - Manure sampling for nutrient analysis: variability and sampling efficacy. AB - Reliable estimation of nutrient concentrations is required to manage animal manure for protecting waters while sustaining crop production. This study was conducted to investigate sample variability and reliable nutrient analysis for several manure types and handling systems. Serial samples were collected from dairy, swine, and broiler poultry operations while manure was being loaded onto hauler tanks or spreaders for field application. Samples were analyzed for total solids (TS), total nitrogen (N), ammoniacal nitrogen (NH4-N), total phosphorus (P), and potassium (K). The least number of samples needed for reliable testing of total N and P, defined as +/- 10% of the experimental means with 99% probability, was obtained for each farm using a computer-intensive random resampling technique. Sample variability within farms, expressed as the coefficient of variation (CV), was mostly 6 to 8% for farms that used agitation of manure storages but several times higher (20-30%) on farms where no agitation was applied during the sampling period. Results from the random resampling procedure indicated that for farms that used agitation, three to five samples were adequate for a representative composite for reliable testing of total N and P; whereas for farms without agitation, at least 40 samples would be required. Data also suggest that using book values for manure nutrient estimations could be problematic because the discrepancies between book standards and measured farm data varied widely from a small amount to several fold. PMID- 11476523 TI - Spatio-temporal patterns of soil phosphorus enrichment in Everglades water conservation area 2A. AB - The Florida Everglades have undergone significant ecological change resulting from anthropogenic manipulation of historical regimes of hydrology, nutrient loading, and fire. Water Conservation Area 2A (WCA-2A) in the northern Everglades has been a focal point for the study of ecological effects of nutrient loading, especially phosphorus (P), from the nearby Everglades Agricultural Area (EAA). The overall objective of our study was to evaluate recent (1990 to 1998) changes in the spatial extent and patterns of soil P enrichment in Everglades WCA-2A. Surface soil was sampled to a depth of 10 cm at 62 sites within WCA-2A during 1998 for analysis of total phosphorus (TP) content. Geostatistical methods were used to create an interpolated grid of soil TP values across WCA-2A. Comparison of the results of this study with a similar study performed in 1990 showed that the extent of soil P enrichment in surface soil and sediments increased between 1990 and 1998, as evidenced by increased coverage of highly P-enriched soil near the primary surface inflows and a general increase in the concentration of soil TP in the interior regions of WCA-2A. Approximately 73% (31 777 ha) of the total land area of WCA-2A was considered P-enriched (soil total P > 500 mg kg(-1)) in 1998, compared with 48% of the land area (20,829 ha) in 1990, an average increase of 1,327 ha yr(-1). Study results indicate that the soil P enrichment "front" has advanced further into the relatively unimpacted interior of WCA-2A during the past several years. PMID- 11476524 TI - The influence of vegetation on sedimentation and resuspension of soil particles in small constructed wetlands. AB - When initiatives to mitigate soil erosion are insufficient or fail, constructed surface flow wetlands (CWs) could be a final buffer to reduce pollution before reaching recipients. The objective of this study was to determine the influence of CW vegetation on the retention of soil particles from arable land. Retention was measured with water flow-proportional sampling systems in the inlet and outlet, sedimentation traps, and sedimentation plates in four small CWs over a period of 5 yr. The surface area of the CWs was 265 to 900 m2, and the average hydraulic loads were 1.2 to 3.4 m d(-1). Watershed areas were 0.5 to 1.5 km2. Annual soil particle retention was 30 to 80% or 14 to 121 kg m(-2). Results show that macrophytes stimulate sediment retention by mitigating resuspension of CW sediment. Five years after construction, resuspension had decreased approximately 40% and was negligible. As vegetation cover increases, the influence of macrophytes on soil particle retention reaches a level where other factors, such as hydraulic load and sediment load, were more important. Macrophytes increased the hydraulic efficiency by reducing short-circuit or preferential flow. However, vegetation did not have any influence on the clay concentration in the sediment. Hence, a possible stimulation of particle flocculation was not detected. Vegetation makes it possible to use the positive effect of a short particle settling distance in shallow ponds by hindering resuspension. PMID- 11476525 TI - Chemical analysis of soil and leachate from experimental wetland mesocosms lined with coal combustion products. AB - Small-scale (1 m2) wetland mesocosm experiments were conducted over two consecutive growing seasons to investigate the effects on soil and leachate chemistry of using a recycled coal combustion product as a liner. The coal combustion product used as a liner consisted of flue gas desulfurization (FGD) by products and fly ash. This paper provides the chemical characteristics of mesocosm soil and leachate after 2 yr of experimentation. Arsenic, Ca, and pH were higher in FGD-lined mesocosm surface soil relative to unlined mesocosms. Aluminum was higher in the soils of unlined mesocosms relative to FGD-lined mesocosms. No significant difference of potentially phytotoxic B was observed between lined and unlined mesocosms in the soil. Higher pH, conductivity, and concentrations of Al, B, Ca, K, and S (SO4-S) were observed in leachate from lined mesocosms compared with unlined controls while Fe, Mg, and Mn were higher in leachate from unlined mesocosms. Concentrations of most elements analyzed in the leachate were below national primary and secondary drinking water standards after 2 yr of experimentation. Initially high pH and soluble salt concentrations measured in the leachate from the lined mesocosms may indicate the reason for early effects noted on the development of wetland vegetation in the mesocosms. PMID- 11476526 TI - Removal and distribution of iron, manganese, cobalt, and nickel within a Pennsylvania constructed wetland treating coal combustion by-product leachate. AB - A flow-through wetland treatment system was constructed to treat coal combustion by-product leachate from an electrical power station at Springdale, Pennsylvania. In a nine-compartment treatment system, four cattail (Typha latifolia L.) wetland cells (designated Cells 1 through 4) successfully removed iron (Fe) and manganese (Mn) from the inlet water; Fe and Mn concentrations were decreased by an average of 91% in the first year (May 1996-May 1997), and by 94 and 98% in the second year (July 1997-June 1998), respectively. Cobalt (Co) and nickel (Ni) were decreased by an average of 39 and 47% in the first year, and 98 and 63% in the second year, respectively. Most of the metal removed by the wetland cells was accumulated in sediments, which constituted the largest sink. Except for Fe, metal concentrations in the sediments tended to be greater in the top 5 cm of sediment than in the 5- to 10- or 10- to 15-cm layers, and in Cell 1 than in Cells 2, 3, and 4. Plants constituted a much smaller sink for metals; only 0.91, 4.18, 0.19, and 0.38% of the Fe, Mn, Co, and Ni were accumulated annually in the aboveground tissues of cattail, respectively. A greater proportion of each metal (except Mn) was accumulated in cattail fallen litter and submerged Chara (a macroalga) tissues, that is, 2.81, 2.75, and 1.05% for Fe, Co, and Ni, respectively. Considerably higher concentrations of metals were associated with cattail roots than shoots, although Mn was a notable exception. PMID- 11476527 TI - Phosphorus sorption characteristics of estuarine sediments under different redox conditions. AB - Phosphorus (P) plays a major role in eutrophication of aquatic systems. Estuarine sediments could function as sources or sinks for P to the overlying water column depending upon their physico-chemical characteristics. Understanding of P sorption phenomena in estuarine sediments is important in regulating the P availability in estuaries. Phosphorus sorption characteristics of sediments from the Indian River Lagoon, Florida, USA, were determined to examine the role of selected physico-chemical properties of the sediments on soluble reactive P status in estuary water. Mean equilibrium P concentrations (EPCo) of 0.75 mg L( 1) and mean P sorption maxima (Smax) of 32.2 mg kg(-1) were obtained under anaerobic conditions, compared with EPCo of 0.05 mg L(-1) and Smax of 132.7 mg kg(-1) under aerobic conditions. The higher EPCo values under anaerobic conditions and the greater Smax values under aerobic conditions were associated with amorphous and poorly crystalline iron. These results suggest that sediments enriched with amorphous and poorly crystalline forms of iron act as an excellent reservoir for P by adsorbing excessive P in aerobic sediment zones and releasing it upon burial under anaerobic conditions. This study also indicates that P compounds in sediments independently maintain equilibrium with P in solutions. Thus, heterogeneous systems like soil and sediment simply behave as a mixture of homogeneous surfaces as far as their P sorption characteristics are concerned, and hence can be successfully described by the Langmuir and Freundlich models. PMID- 11476528 TI - Malaria deaths following inappropriate malaria chemoprophylaxis--United States, 2001. AB - During January-March 2001, two U.S. citizens died from malaria after taking chloroquine alone or with proguanil for malaria chemoprophylaxis in countries with known chloroquine-resistant Plasmodium falciparum malaria. Chloroquine containing chemoprophylaxis regimens are not recommended by CDC for persons traveling to areas with known chloroquine-resistant P. falciparum. This report summarizes the investigation of the two cases and underscores the need for clinicians and travelers to know the recommended options for malaria chemoprophylaxis when traveling to locations with chloroquine-resistant malaria. PMID- 11476529 TI - Evaluation of a regional pilot program to prevent mother-infant HIV transmission- Thailand, 1998-2000. AB - Worldwide, approximately 2.2 million women and 600,000 infants are infected with human immunodeficiency virus (HIV) each year. Extended zidovudine prophylaxis and other antiretroviral and obstetric interventions and the avoidance of breast feeding have reduced dramatically mother-infant HIV transmission in countries with adequate health-care resources. However, in developing countries, where the impact of HIV is greatest, implementation has been limited by the complexity and expense of these interventions. In Thailand, where approximately 15,000 infants are born to HIV-infected women each year, the Ministry of Public Health (MOPH) has collaborated with other organizations to identify simpler and more cost effective interventions to reduce mother-infant HIV transmission. In 1998, a placebo-controlled clinical trial in Thailand using a simplified zidovudine regimen from 36 weeks' gestation until delivery reduced the risk for mother infant transmission by 50%. In 1998, MOPH initiated a pilot program to prevent mother-infant HIV transmission in region 7, a rural area in northeastern Thailand with an antenatal HIV prevalence of approximately 1%, to assess program feasibility, effectiveness, and acceptability. This report summarizes an evaluation of the 2-year pilot program, which indicated that acceptance of HIV testing and adherence to zidovudine were high and HIV transmission was reduced. The findings demonstrate the feasibility of implementing programs to prevent mother-infant HIV transmission on a large scale in a developing country. PMID- 11476530 TI - Hantavirus pulmonary syndrome--Vermont, 2000. AB - In 1993, an outbreak of an unexplained pulmonary illness occurred in the southwestern United States. This outbreak led to the first description of hantavirus pulmonary syndrome (HPS), a rodentborne hantaviral infection. Hantaviruses have been found in rodents in rural areas throughout the United States, but most infection has occurred in the southwest. This report describes the first HPS case in Vermont and underscores the importance of preventing exposure to peridomestic rodents and recognizing the signs and symptoms of HPS. PMID- 11476531 TI - Wound irrigation in musculoskeletal injury. AB - Wound irrigation to remove debris and lessen bacterial contamination is an essential component of open fracture care. However, considerable practice variation exists in the details of technique. Volume is an important factor; increased volume improves wound cleansing to a point, but the optimal volume is unknown. High-pressure flow has been shown to remove more bacteria and debris and to lower the rate of wound infection compared with low-pressure irrigation, although recent in vitro and animal studies suggest that it may also damage bone. Pulsatile flow has not been demonstrated to increase efficacy. Antiseptic additives can kill bacteria in the wound, but host-tissue toxicities limit their use. Animal and clinical studies of the use of antiseptics in contaminated wounds have yielded conflicting outcomes. Antibiotic irrigation has been effective in experimental studies in some types of animal wounds, but human clinical data are unconvincing due to poor study design. There are few animal or clinical studies of musculoskeletal wounds. Detergent irrigation aims to remove, rather than kill, bacteria and has shown promise in animal models of the complex contaminated musculoskeletal wound. PMID- 11476532 TI - Muscle contusion injuries: current treatment options. AB - Muscle contusion is second only to strain as the leading cause of morbidity from sports-related injuries. Severity depends on the site of impact, the activation status of the muscles involved, the age of the patient, and the presence of fatigue. The diagnosis has traditionally been one of clinical judgment; however, newer modalities, including ultrasonography, magnetic resonance imaging, and spectroscopy, are becoming increasingly important in both identifying and delineating the extent of injury. Although controlled clinical studies are scarce, animal research into muscle contusions has allowed the description of the natural healing process, which involves a complex balance between muscle repair, regeneration, and scar-tissue formation. Studies are being performed to evaluate the effects of anti-inflammatory medications, corticosteroids, operative repair, and exercise protocols. Prevention and treatment of complications such as myositis ossificans have also been stressed, but recognition may improve the outcome of these ubiquitous injuries. PMID- 11476533 TI - "Floating knee" injuries: ipsilateral fractures of the femur and tibia. AB - Ipsilateral fractures of the femur and tibia have been called "floating knee" injuries and may include combinations of diaphyseal, metaphyseal, and intra articular fractures. These are often high-energy injuries and most frequently occur in the polytrauma patient. Many of these fractures are open, with associated vascular injuries. Surgical stabilization of both fractures and early mobilization of the patient and the extremity produce the best clinical outcomes. The use of a radiolucent operating room table and the introduction of retrograde intramedullary fixation of femoral fractures have facilitated surgical stabilization of some floating-knee fracture patterns. Although treatment planning for each fracture in the extremity should be considered individually to achieve the optimal result, the effect of that decision must be considered in light of the overall injury status of the entire extremity. Collateral ligament and meniscal injuries may also be associated with this fracture complex. Complications (such as compartment syndrome, loss of knee motion, failure to diagnose knee ligament injury, and the need for amputation) are not infrequent. Better results and fewer complications are observed when both fractures are diaphyseal than when one or both are intra-articular. PMID- 11476534 TI - Trigger digits: diagnosis and treatment. AB - Stenosing tenosynovitis of the thumb and fingers is a very common problem seen by the primary-care physician, the orthopaedic surgeon, and the hand surgeon. Primary stenosing tenosynovitis is usually idiopathic and occurs more frequently in middle-aged women than in men, but can be seen even in infancy. Secondary stenosing tenosynovitis of the digits can occur in patients with rheumatoid arthritis, diabetes mellitus, gout, and other disease entities that cause connective tissue disorders. The diagnosis of triggering digits is generally not subtle and can be made on the basis of an adequate clinical examination. Classification according to the type of tenosynovitis and the time from onset of symptoms may be prognostically significant and may also affect the treatment outcome. As many as 85% of triggering fingers and thumbs can be treated successfully with corticosteroid injections and nonsteroidal anti-inflammatory drugs. Surgical release is generally indicated when nonoperative treatment fails. Percutaneous A1 pulley release can now be performed safely as an office procedure. PMID- 11476535 TI - Arterial complications and total knee arthroplasty. AB - Arterial complications after total knee arthroplasty (TKA) are rare; however, the sequelae can be disastrous. Infection and the need for amputation or vascular reconstructive surgery are not uncommon. A thorough preoperative assessment can identify at-risk patients, many, if not all, of whom have preexisting peripheral arterial disease. In the presence of peripheral arterial disease, the use of a tourniquet during TKA has been implicated in subsequent arterial complications. Following the guidelines that have been established regarding preoperative assessment, the role of the vascular surgeon, and the use of a tourniquet before and during TKA can assist the orthopaedic surgeon in assessing candidates for TKA and reducing the risk of arterial complications. PMID- 11476536 TI - Diffuse idiopathic skeletal hyperostosis: musculoskeletal manifestations. AB - Diffuse idiopathic skeletal hyperostosis (DISH) is a common disorder of unknown etiology that is characterized by back pain and spinal stiffness. There may be mild pain if ankylosis has occurred. The condition is recognized radiographically by the presence of "flowing" ossification along the anterolateral margins of at least four contiguous vertebrae and the absence of changes of spondyloarthropathy or degenerative spondylosis. Even in patients who present with either lumbar or cervical complaints, radiographic findings are almost universally seen on the right side of the thoracic spine. Thus, radiographic examination of this area is critical when attempting to establish a diagnosis of DISH. The potential sequelae of hyperostosis in the cervical and lumbar spine include lumbar stenosis, dysphagia, cervical myelopathy, and dense spinal cord injury resulting from even minor trauma. There may be a delay in diagnosis of spinal fractures in a patient with DISH because the patient often has a baseline level of spinal pain and because the injury may be relatively trivial. The incidence of delayed neurologic injury due to such fractures is high as a result of unrecognized instability and subsequent deterioration. Extraspinal manifestations are also numerous and include an increased risk of heterotopic ossification after total hip arthroplasty. Prophylaxis to prevent heterotopic ossification may be indicated for these patients. PMID- 11476537 TI - Pediatric ankle fractures: evaluation and treatment. AB - Pediatric ankle fractures account for approximately 5% of pediatric fractures and 15% of physeal injuries. The biomechanical differences between mature and immature bones, as well as the differing forces applied to those bones, help explain the differences between adult and pediatric fractures. The potential complications associated with pediatric ankle fractures include those seen with adult fractures (such as posttraumatic arthritis, stiffness, and reflex sympathetic dystrophy) as well as those that result from physeal damage (including leg-length discrepancy, angular deformity, or a combination thereof). The goals of treatment are to achieve and maintain a satisfactory reduction and to avoid physeal arrest. A knowledge of common pediatric ankle fracture patterns and the pitfalls associated with their evaluation and treatment will aid the clinician in the effective management of these injuries. PMID- 11476538 TI - Colorimetric silver detection of DNA microarrays. AB - Development of microarrays has revolutionized gene expression analysis and molecular diagnosis through miniaturization and the multiparametric features. Critical factors affecting detection efficiency of targets hybridization on microarray are the design of capture probes, the way they are attached to the support, and the sensitivity of the detection method. Microarrays are currently detected in fluorescence using a sophisticated confocal laser-based scanner. In this work, we present a new colorimetric detection method which is intented to make the use of microarray a powerful procedure and a low-cost tool in research and clinical settings. The signal generated with this method results from the precipitation of silver onto nanogold particles bound to streptavidin, the latter being used for detecting biotinylated DNA. This colorimetric method has been compared to the Cy-3 fluorescence method. The detection limit of both methods was equivalent and corresponds to 1 amol of biotinylated DNA attached on an array. Scanning and data analysis of the array were obtained with a colorimetric-based workstation. PMID- 11476539 TI - Automatic Edman microsequencing of peptides containing multiple unnatural amino acids. AB - It is now routine using automatic Edman microsequencing to determine the primary structure of peptides or proteins containing natural amino acids; however, a deficiency in the ability to readily sequence peptides containing unnatural amino acids remains. With the advent of synthetic peptide chemistry, combinatorial chemistry, and the large number of commercially available unnatural amino acids, there is a need for efficient and accurate structure determination of short peptides containing many unnatural amino acids. In this study, 35 commercially available alpha-unnatural amino acids were selected to determine their elution profile on an ABI protein sequencer. Using a slightly modified gradient program, 19 of these 35 PTH amino acids can be readily resolved and distinguished from common PTH amino acids at low picomole levels. These unnatural amino acids in conjunction with the 20 natural amino acids can be used as building blocks to construct peptide libraries, and peptide beads isolated from these libraries can be readily microsequenced. To demonstrate this, we synthesized a simple tripeptide "one-bead one-compound" combinatorial library containing 14 unnatural and 19 natural amino acids and screened this library for streptavidin-binding ligands. Microsequencing of the isolated peptide-beads revealed the novel motif Bpa-Phe(4-X)-Aib, wherein X = H, OH, and CH3. PMID- 11476540 TI - Ribosomal 18S RNA prevails over glyceraldehyde-3-phosphate dehydrogenase and beta actin genes as internal standard for quantitative comparison of mRNA levels in invasive and noninvasive human melanoma cell subpopulations. AB - The comparison of the gene expression profiles between two subpopulations of melanoma cells (1C8 and T1C3) derived from the tumor of one patient by cDNA array revealed differences in GAPDH and beta-actin gene levels. These two housekeeping genes were up-regulated in invasive T1C3 melanoma cells compared to noninvasive 1C8 cells. Since cDNA array results were not confirmed by conventional RT-PCR throughout the exponential phase of amplification, we performed duplex relative RT-PCR using ribosomal 18S RNA as internal standard including competimer technology. Statistical analyses provided significant evidence that invasive T1C3 melanoma cells exhibited a twofold higher mRNA level of both GAPDH and beta-actin than noninvasive 1C8 cells. This study demonstrates that the duplex relative RT PCR procedure including ribosomal 18S RNA as internal standard and competimer technology is precise for RNA quantification and is tailored for cDNA array validation. Our data provide molecular evidence that cellular subpopulations of the same pathological origin are highly heterogeneous and extend the concept that the selection of an appropriate internal control for comparative mRNA analysis should be adapted to each model of human cancers. PMID- 11476541 TI - Dual labeling of a binding protein allows for specific fluorescence detection of native protein. AB - Fluorescence resonance energy transfer has been investigated in the context of specific detection of unlabeled proteins. A model system based on the staphylococcal protein A (SPA)-IgG interaction was designed, in which a single domain was engineered to facilitate site-specific incorporation of fluorophores. An Asn23Cys mutant of the B domain from SPA was expressed in Escherichia coli and subsequently labeled at the introduced unique thiol and at an amino group, using N-iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine (1,5-IAEDANS) and succinimidyl 6-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoate (NBD-X, SE), respectively. Biosensor analysis of purified doubly labeled protein showed that high-affinity binding to the Fc region of IgG was retained. The fluorescence emission spectrum of the doubly labeled protein showed a shift in the relative emission of the two fluorophores in the presence of Fc3(1) fragments, which bind specifically to the B domain. In addition, the fluorescence emission ratio 480/525 nm was shown to increase with increasing concentration of Fc3(1), whereas the presence of a control protein did not affect the emission ratio over the same concentration range. PMID- 11476542 TI - Quantification of sodium dodecyl sulfate in microliter-volume biochemical samples by visible light spectroscopy. AB - A method for sodium dodecyl sulfate (SDS) quantitation in microliter-volume complex biochemical samples is described. The quantitation is based on the use of a dye, stains-all, the color of which changes from intense fuchsia to yellow upon addition of SDS. We show that this color change is gradual and proportional to the amount of SDS added to the stains-all solution, thus allowing its use to reliably quantitate SDS in biochemical samples by means of a visible light spectrophotometer. A large number of compounds widely used in biochemistry are herein shown not to interfere with the SDS measurement when they are present in the sample at usual biochemical concentrations. Furthermore, linearity between the color change and the amount of SDS present in the sample is never impaired when huge amounts of these compounds are also present, thus making this quantitation method highly reliable with use of a calibration curve. The method allows easy and reliable quantitation of microgram amounts of SDS in microliter volume biochemical samples. PMID- 11476543 TI - Continuous recording of long-chain acyl-coenzyme a synthetase activity using fluorescently labeled bovine serum albumin. AB - The fluorescence-based long-chain fatty acid probe BSA-HCA (bovine serum albumin labeled with 7-hydroxycoumarin-4-acetic acid) is shown to respond to binding of long-chain acyl-CoA thioesters by quenching of the 450 nm fluorescence emission. As determined by spectrofluorometric titration, binding affinities for palmitoyl , stearoyl-, and oleoyl-CoA (Kd = 0.2-0.4 microM) are 5-10 times lower than those for the corresponding nonesterified fatty acids. In the presence of detergent (Chaps, Triton X-100, n-octylglucoside) above the critical micelle concentration, acyl-CoA partitions from BSA-HCA and into the detergent micelles. This allows BSA HCA to be used as a fluorescent probe for continuous recording of fatty acid concentrations in detergent solution with little interference from acyl-CoA. Using a calibration of the fluorescence signal with fatty acids in the C14 to C20 chain-length range, fatty acid consumption by Pseudomonas fragi and rat liver microsomal acyl-CoA synthetase activities are measured down to 0.05 microM/min with a data sampling rate of 10 points per second. This new method provides a very promising spectrofluorometric approach to the study of acyl-CoA synthetase reaction kinetics at physiologically relevant (nM) aqueous phase concentrations of fatty acid substrates and at a time resolution that cannot be obtained in isotopic sampling or enzyme-coupled assays. PMID- 11476544 TI - Identification and location of a cysteinyl posttranslational modification in an amyloidogenic kappa1 light chain protein by electrospray ionization and matrix assisted laser desorption/ionization mass spectrometry. AB - Amyloid-deposited light chain (AL) amyloidosis is correlated with the overproduction of a monoclonal immunoglobulin light chain protein by a B lymphocyte clone. Since the amyloid fibril deposits in AL amyloidosis most often consist of the N-terminal fragments of the light chain, the majority of studies have focused on the determination of the primary structure of the protein, and reducing agents have been used routinely in the initial purification process. In this study, two light chain proteins were isolated and purified, without reduction, from the urine of a patient diagnosed with kappa 1 (kappa1) AL amyloidosis. One protein had a relative molecular mass of 12,000 and the other 24,000. Electrospray ionization and matrix-assisted laser desorption/ionization mass spectrometry, in combination with enzymatic digestions, were used to verify the amino acid sequences and identify and locate posttranslational modifications in these proteins. The 12-kDa protein was confirmed to be the N-terminal kappa1 light chain fragment (variable region) consisting of residues 1-108 or 1-109 and having one disulfide bond. The 24-kDa protein was determined to be the intact kappa1 light chain containing a cysteinyl posttranslational modification at Cys214 and disulfide bonds located at Cys23-Cys88, Cys134-Cys194, and Cys214-Cys. The methods used in this report enable high-sensitivity determination of amino acid sequence and variation in intact and truncated light chains as well as posttranslational modifications. This approach facilitates consideration of the effect of cysteinylation on the native protein structure and the potential involvement of this modification in AL amyloidosis. PMID- 11476545 TI - Analysis of antibody-antigen interactions in mixtures containing reactive and nonreactive components using size-exclusion high-performance (pressure) liquid chromatography. AB - Analysis of antibody-antigen interactions using size-exclusion high-performance (pressure) liquid chromatography was applied to a polyvalent system composed of both reactive and nonreactive components. Mixtures containing varying concentrations of antivenin (Crotalidae) polyvalent (equine origin) and either Crotalus atrox (Western diamondback rattlesnake) venom (CV) or isolated C. atrox phospholipase A2 (PLA2) were separated using a Bio-Sil SEC 250-5 size-exclusion column (300 x 7.8 mm). Major regions containing high-molecular-weight aggregates, antivenin IgG, and CV components or PLA2 could be identified from the elution profiles. Changes in elution profile areas could be modeled by equations derived from the law of mass action that included values for the maximum fraction of reactivity, antigen valence, apparent binding constants, profile area proportionality constants, and nonreactive profile area. The analysis was simple, fast, and readily interpretable and may be applicable to a variety of situations in which stable antigen-antibody complexes are formed in the presence of nonreactive components. PMID- 11476546 TI - Preparation and evaluation of Ricinus communis agglutinin affinity adsorbents using polymeric supports. AB - A practicable and efficient procedure for preparation of Ricinus communis agglutinin (RCA) affinity adsorbents has been developed. For immobilization of RCA two different polymer-based supports, Toyopearl and TSKgel (TosoHaas), were used. RCA has been successfully immobilized onto these supports with amounts of coupled ligand between 15 and 23 mg/g dry support and corresponding coupling yields of 69-93% (w/w). The prepared affinity adsorbents were characterized concerning their binding capacity for the glycoprotein asialofetuin (ASF) and accessibility of the ligand binding sites. The high accessibility of 80% showed that steric hindrance was negligible at the present ligand density. RCA-Toyopearl was successfully applied in affinity chromatography of glycoproteins indicating its high specificity. A long-term stability test proved no change in capacity for a period of at least 12 months. High-performance affinity chromatography (HPLAC) was carried out using RCA-TSKgel. Experimental results showed that the prepared adsorbents are suitable for selective separation of glycoproteins and oligosaccharides and therefore can be used for investigations of adsorption characteristics of glycoconjugates and for laboratory-scale preparations. PMID- 11476547 TI - Surface phosphophilicity of aluminum-containing adjuvants probed by their efficiency for catalyzing the P--O bond cleavage with chromogenic and fluorogenic substrates. AB - Aluminum-containing adjuvants are widely used in a variety of vaccine products, such as recombinant proteins, virus-like particles, conjugated polysaccharides, and recently DNA vaccines. Aluminum-containing adjuvants are also known to have a high affinity to inorganic phosphate and its mono- or diesters. Since phosphate groups are present in many antigens as well as the natural physiological environment, a better understanding of the interactions between phosphate and phospho-containing species could help in the design of improved vaccines. This report describes a convenient and novel continuous procedure to measure the avidity denoted by the new term "phosphophilicity" of phosphate and phosphate esters to the surface of aluminum-containing adjuvants. The assay measures the rate of hydrolysis of a fluorogenic substrate-6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP)-with a microplate reader. This method was based on the fundamental bioorganic phenomenon that when a tight binding event occurs, the effective concentration of nucleophile(s) will be significantly increased in the proximity of the P atom for a nucleophilic reaction (i.e., the cleavage of the P&bond;O bond) to take place. A very good leaving group (pK(a) of DiFMU approximately 4.7) in the phosphate monoester substrate makes the assay highly sensitive. Top reading of the nascent fluorescence makes the assay very convenient with no need to separate the particulate adjuvants from the reaction mixtures. The results from this assay are consistent with catalysis of the chromogenic phosphate mono- or diesters. PMID- 11476548 TI - A spectrophotometric assay for quantitative determination of kcat of herpes simplex virus type 1 thymidine kinase substrates. AB - A simple method to determine the in vitro catalytic turnover constant of several substrates of herpes simplex virus type 1 thymidine kinase is presented in this study. The method is based on a continuous spectroscopic enzyme-coupled assay and allows one to monitor the herpes simplex virus type 1 thymidine kinase activity in the presence of unlabeled substrates. A clear correlation between the catalytic turnover constant and the rate of decrease in absorbance over time during the assay has been demonstrated. Exploiting this correlation, this method has been used to determine rapidly and precisely the catalytic turnover constant of antiviral lead compounds not readily available in the radioactive labeled form. PMID- 11476549 TI - Quantitative analysis and molecular species fingerprinting of triacylglyceride molecular species directly from lipid extracts of biological samples by electrospray ionization tandem mass spectrometry. AB - Herein we describe a rapid, simple, and reliable method for the quantitative analysis and molecular species fingerprinting of triacylglycerides (TAG) directly from chloroform extracts of biological samples. Previous attempts at direct TAG quantitation by positive-ion electrospray ionization mass spectrometry (ESI/MS) were confounded by the presence of overlapping peaks from choline glycerophospholipids requiring chromatographic separation of lipid extracts prior to ESI/MS analyses. By exploiting the rapid loss of phosphocholine from choline glycerophospholipids, in conjunction with neutral-loss scanning for individual fatty acids, overlapping peaks in the ESI mass spectrum were deconvoluted generating a detailed molecular species fingerprint of individual TAG molecular species directly from chloroform extracts of biological samples. This method readily detects as little as 0.1 pmol of each TAG molecular species from chloroform extracts and is linear over a 1000-fold dynamic range. The sensitivity of individual TAG molecular species to ESI/MS/MS analyses correlated with the unsaturation index and inversely correlated with total aliphatic chain length of TAG. An algorithm was developed which identifies sensitivity factors, thereby allowing the rapid quantitation and molecular species fingerprinting of TAG molecular species directly from chloroform extracts of biological samples. PMID- 11476550 TI - A competitive fluorescence assay to measure the reactivity of compounds. AB - A sensitive competitive method was developed for assessing the reactivity of compounds toward glutathione and toward thiols in general. The method employs the reaction of the fluorogenic reagent fluorescein-5-maleimide (FM) with glutathione (GSH) to generate a large increase in fluorescence emission. When the reaction is measured in the presence of a compound that competes with FM toward GSH, the rate constant for fluorescent product formation increases while the total amount of product formed at the end of the reaction decreases. These changes in the presence of a series of competitor concentrations allow one to calculate the rate constant of the reaction of the competitor with GSH. At 23 degrees C, pH 7.40 in PBS buffer the second-order rate constant of the FM-GSH reaction is k2 = (1.67 +/ 0.32) x 10(4) M(-1) x s(-1). Two GSH-reactive compounds were evaluated: the second-order rate constant for the reaction of PNU-27707 with GSH under our experimental conditions is k(i) = 5660 +/- 266 M(-1) x s(-1), while that of PNU 37802 is k(i) = 21,200 +/- 1600 M(-1) x s(-1). The method is easily adaptable to a high-throughput screening format. PMID- 11476551 TI - Biophysical evidence of arm-domain interactions in AraC. AB - We report development of a method for the direct measurement of the interaction between the N-terminal arm and the remainder of the dimerization domain in the Escherichia coli AraC protein, the regulator of the l-arabinose operon. The interaction was measured using surface plasmon resonance to monitor the association between the immobilized peptide arm and the dimerization domain, truncated of its arm, in solution. As expected from genetic and physiological data, the interaction is strongly stimulated by l-arabinose and is insensitive to sugars like d-glucose or d-galactose. Alterations in the sequence of the arm which physiological experiments predict either to strengthen or weaken the arm produce the expected responses. PMID- 11476552 TI - One-step duplex reverse transcription-polymerase chain reaction for quantitative assessment of RNA degradation. PMID- 11476553 TI - One-step extraction of RNA from archival biopsies. PMID- 11476554 TI - A spectrophotometric assay of arginase. PMID- 11476555 TI - Enhanced protein recovery and reproducibility from pull-down assays and immunoprecipitations using spin columns. PMID- 11476556 TI - An assay for phosphoinositide phosphatases utilizing fluorescent substrates. PMID- 11476557 TI - Epidemiology of endometrial cancer. AB - Endometrial cancer is the commonest gynaecological cancer mostly affecting women in the post-menopausal age group. Rates vary worldwide and are highest in white women in Western populations. Some risk factors are related to reproduction, such as early age at menarche, late age at menopause and nulliparity, while others are more directly oestrogen-related, for example, conditions such as the polycystic ovarian syndrome. Use of unopposed oestrogen replacement therapy is associated with an increased risk, and use of the combined oral contraceptive pill is associated with a decreased risk. The relationship between tamoxifen and endometrial cancer is not established. Obesity, diabetes and hypertension increase the risk of endometrial cancer while smoking, low-fat diets and physical exercise appear to decrease the risk; all of these possibly exert their effects by various indirect influences on oestrogen levels, thus influencing the level of stimulation of the target endometrial epithelium. PMID- 11476558 TI - Molecular genetics and endometrial cancer. AB - Endometrial cancer is the ninth most common malignancy in females. Inherited forms of this malignancy exist. Mutations in mismatch repair genes result in hereditary non-polyposis colorectal cancer, which confers a lifetime risk of bowel cancer between 60-80% and an endometrial cancer risk of up to 60%. Current screening involves the use of transvaginal ultrasound and hysteroscopy. Genetic testing for mutations in the mismatch repair genes is available, and if a pathogenic change is found within a family, predictive testing becomes available for unaffected family members. If blood samples from family members are unavailable, tumour blocks may be studied to assess microsatellite instability, a feature of mismatch repair gene mutations. While mutations in the mismatch repair genes are found in inherited endometrial cancer they are rarely seen in sporadic cancers. However, there are a range of somatic gene mutations that are currently being studied in order to provide insight into the pathogenesis of endometrial cancer. PMID- 11476559 TI - Tamoxifen, screening and new oestrogen receptor modulators. AB - Tamoxifen is a selective oestrogen receptor modulator (SERM) with anti oestrogenic properties in the breast and oestrogenic effects in tissues such as bone and the cardiovascular system. It is an excellent breast cancer drug for all stages of the disease. Its SERM profile makes it a valuable alternative to hormone replacement therapy, especially for women at high risk of breast cancer. Tamoxifen, however, increases the incidence of benign and malignant lesions of the uterus. Secondary prevention of these, early detection and treatment, is feasible but not cost-effective in breast cancer patients because potential endometrial risks do not outweigh beneficial effects in the breast. This may be different in healthy women with an as yet unknown benefit on breast cancer mortality. We review the literature on the importance of tamoxifen's endometrial lesions and balance available evidence on whether and how best to screen them. In a subset of tamoxifen users it seems advisable to assess the uterine cavity prior to intake with a yearly endometrial assessment as pointed out, starting 3 years after initiation of treatment. In most cases there is endometrial thickening on ultrasonographic assessment and additional tests such as hydrosonography or hysteroscopy are required to confirm an empty atrophic uterus as remains the case in most asymptomatic women on tamoxifen. Newer compounds, such as raloxifene, have a similar SERM profile to tamoxifen but are neutral on the uterus. This has recently been proven by 3 years of endometrial follow-up data. Longer endometrial safety will hopefully confirm these early findings. Whether other SERMs in development are better, and which of them is better for the breast, is to be demonstrated in ongoing studies. PMID- 11476560 TI - Ultrasound, hysteroscopy and endometrial biopsy in the investigation of endometrial cancer. AB - Over the course of the last two decades hysteroscopy with endometrial biopsy has begun to replace dilation and curettage as the method of choice for the diagnosis of endometrial carcinoma. In the majority of women this can be performed as an outpatient procedure with no loss in diagnostic accuracy. Transvaginal ultrasound measurement of endometrial thickness provides a highly sensitive and less invasive alternative means of assessing the endometrium but has a low positive predictive value for cancer, especially in women taking hormone replacement therapy. The cut-off value used to define normality needs to take into account patient age and ethnic origin. Ultrasound screening may not be suitable for women taking tamoxifen and those with recurrent or late-onset abnormal uterine bleeding. PMID- 11476561 TI - Optimal management of endometrial hyperplasia. AB - The optimal management of endometrial hyperplasia is the subject of considerable debate. In this chapter the development of our current classification of endometrial hyperplasias is outlined in some detail in order to give an understanding of the complexity of the problem of determining the malignant potential of the hyperplasia which is the central issue in determining optimal treatment. While hysterectomy is still the definitive treatment for older women with hyperplasia, conservative therapy is perfectly acceptable in a defined group of younger women who are closely monitored. PMID- 11476562 TI - Surgery for endometrial cancer: what type and by whom? AB - Endometrial cancer has far too long been regarded as a simple disease to treat. As such it has generally remained in the hands of the generalist obstetrician/gynaecologist. In order to optimize the choice of surgery, careful pre-operative evaluation is essential with respect to pathology of a biopsy, radiological assessment of extent of disease and evaluation of fitness for anaesthesia. The standard procedure is a total abdominal hysterectomy and bilateral salpingo-oophorectomy. However, consideration should be given to pelvic lymphadenectomy in high-risk cases. Surgery for high-risk endometrial cancer should be performed by gynaecological oncologists. PMID- 11476563 TI - Radiation therapy in endometrial cancer. AB - The role of radiation in endometrial cancer, especially in the adjuvant setting, is controversial. Factors that influence radiotherapy recommendations include surgical considerations, pathological findings, potential sites of disease recurrence and the practice philosophies of the individual physician. It has been demonstrated that adjuvant radiotherapy following primary surgery significantly improves pelvic tumour control, but has no measurable impact on overall survival in an unselected patient population. Studies to date have been hampered by the inclusion of patients with a wide spectrum of prognostic features; this may decrease the likelihood of observing greater benefit in discriminate subsets at higher risk of relapse. Further trials are required to define clinical prognosis more precisely and to investigate the role of radiation in higher-risk patients. In the meantime, we propose guidelines for radiotherapy in endometrial cancer which serve as bases for discussion and collaboration among physicians and as platforms for future study and progress. PMID- 11476564 TI - Clear-cell and papillary serous cancer: treatment options. AB - Clear-cell carcinoma (CCC) and serous papillary carcinoma of the endometrium (UPSC) are rare subtypes of endometrial carcinoma (10%). The histological diagnosis can be made on the dilation and curettage specimens in both types in a very high percentage of the cases. This is important in the planning of treatment. CCC and UPSC are associated with about 50% of all relapses occurring in endometrial carcinoma, and the 5-year survival rate is, on average, 42% and 27% respectively. Surgico-pathological stage, age, and vessel invasion are independent prognostic factors for both groups. The recurrence rate is extremely high, and the most frequent extra-pelvic sites of relapse are the upper abdomen, lungs and liver. Stage Ia patients treated with complete surgical staging alone have a low risk of relapse and need not be offered adjuvant systemic therapy or pelvic radiation. The treatment of patients with CCC and UPSC stage Ib, Ic, II and III should include radical debulking surgery and some form of adjuvant therapy, but it is not clear which type is most effective. Adjuvant pelvic radiotherapy plus intracavitary radiotherapy is usually given in early-stage disease and pelvic radio therapy/or whole abdomen irradiation plus adjuvant systemic chemotherapy (PAC) in advanced disease. However, we are urgently waiting for a large prospective randomized study to compare both modalities. Paclitaxel, alone or in combination, is currently being tested in phase II studies. PMID- 11476565 TI - Is there any place for cytotoxic chemotherapy in endometrial cancer? AB - Cytotoxic chemotherapy has an established role in the treatment of many solid tumours that are considered to be incurable with any modern treatment method. Such treatment may result in an improvement in quality of life without influencing overall survival. In this chapter the evidence to support the use of chemotherapy in patients with advanced or recurrent endometrial adenocarcinoma is reviewed. The most effective single agent and combination treatments are outlined. Although evidence from randomized trials is limited, combination chemotherapy can lead to response rates of over 40% in patients with advanced disease. The role of chemotherapy as adjuvant treatment in patients with early stage disease is less well defined and this treatment is not recommended outside a clinical trial. The role of chemotherapy for treatment of the aggressive histological variant, uterine papillary serous carcinoma is also discussed. PMID- 11476566 TI - Hormonal treatment of endometrial cancer: past, present and future. AB - The concept that hormonal therapy may be useful in the treatment of endometrial cancer antedated the pharmaceutical availability of progestational compounds. By 1959, initial studies demonstrated the ability of progestins to reverse endometrial hyperplasias. Thereafter, progestins and other hormonal agents have been used in various roles as treatment for endometrial cancers. This chapter reviews the use of hormonal agents for the treatment of primary and metastatic/recurrent endometrial cancer, as well as such treatment in an adjuvant setting. Major problems in enhancing the efficacy of endocrine therapy of cancers arising from hormonally responsive tissues are also considered. The regulations of steroid-hormone receptor expression in endometrial and breast cancers continues to be an active area of research interest. PMID- 11476570 TI - Regulation of MAP kinase by the BMP-4/TAK1 pathway in Xenopus ectoderm. AB - Bone morphogenetic protein-4 (BMP-4) induces epidermis and represses neural fate in Xenopus ectoderm. Our previous findings implicate p42 Erk MAP kinase (MAPK) in the response to neural induction. We have examined the effects of BMP-4 on MAPK activity in gastrula ectoderm. Expression of a dominant negative BMP-4 receptor resulted in a 4.5-fold elevation in MAPK activity in midgastrula ectoderm. MAPK activity was reduced in ectoderm expressing a constitutively active BMP-4 receptor, or ectoderm treated with BMP-4 protein in the presence or absence of cycloheximide. Overexpression of TAK1 led to a reduction in MAPK activity in early gastrula ectoderm. The inhibitory effects of TAK1 could be reversed by 1 microM SB 203580, a p38 inhibitor. Treatment of isolated ectoderm with SB 203580 led to expression of otx2, NCAM, and noggin. Western blot analyses indicated that the BMP-4 pathway does not activate JNKs in ectoderm. Our findings indicate that BMP-4 inhibits ectodermal MAPK activity through a TAK1/p38-type pathway. MAPK has been shown to inactivate Smad1. Thus, our results suggest that BMP-4 and MAPK pathways are mutually antagonistic in Xenopus ectoderm, and that interactions between these pathways may govern the choice between epidermal and neural fate. PMID- 11476571 TI - The BMP/CHORDIN antagonism controls sensory pigment cell specification and differentiation in the ascidian embryo. AB - We have investigated the role of the bone morphogenetic protein (BMP) pathway during neural tissue formation in the ascidian embryo. The orthologue of the BMP antagonist, chordin, was isolated from the ascidian Halocynthia roretzi. While both the expression pattern and the phenotype observed by overexpressing chordin or BMPb (the dpp-subclass BMP) do not suggest a role for these factors in neural induction, BMP/CHORDIN antagonism was found to affect neural patterning. Overexpression of BMPb induced ectopic sensory pigment cells in the brain lineages that do not normally form pigment cells and suppressed pressure organ formation within the brain. Reciprocally, overexpressing chordin suppressed pigment cell formation and induced ectopic pressure organ. We show that pigment cell formation occurs in three steps. (1) During cleavage stages ectodermal cells are neuralized by a vegetal signal that can be substituted by bFGF. (2) At the early gastrula stage, BMPb secreted from the lateral nerve cord blastomeres induces those neuralized blastomeres in close proximity to adopt a pigment cell fate. (3) At the tailbud stage, among these pigment cell precursors, BMPb induces the differentiation of specifically the anterior type of pigment cell, the otolith; while posteriorly, CHORDIN suppresses BMP activity and allows ocellus differentiation. PMID- 11476572 TI - The Caenorhabditis elegans Six/sine oculis class homeobox gene ceh-32 is required for head morphogenesis. AB - Caenorhabditis elegans has four members of the Six/sine oculis class of homeobox genes, ceh-32, ceh-33, ceh-34, and ceh-35. Proteins encoded by this gene family are transcription factors sharing two conserved domains, the homeodomain and the Six/sine oculis domain, both involved in DNA binding. ceh-32 expression was detected during embryogenesis in hypodermal and neuronal precursor cells and later in descendants of these cells as well as in gonadal sheath cells. RNAi inactivation studies suggest that ceh-32 plays a role in head morphogenesis, like vab-3, the C. elegans Pax-6 orthologue. ceh-32 and vab-3 are coexpressed in head hypodermal cells and ceh-32 mRNA levels are reduced in vab-3 mutants. Moreover, ectopic expression of VAB-3 in transgenic worms is able to induce ceh-32 ectopically. In addition, we demonstrate that VAB-3 is able to bind directly to the ceh-32 upstream regulatory region in vitro and to activate reporter gene transcription in a yeast one-hybrid system. Our results suggest that VAB-3 acts upstream of ceh-32 during head morphogenesis and directly induces ceh-32. Thus, ceh-32 appears to be the first target gene of VAB-3 identified so far. PMID- 11476573 TI - A possible role for the cnidarian homologue of serum response factor in decision making by undifferentiated cells. AB - We have isolated the serum response factor (SRF) homologue from two hydrozoans, the freshwater polyp Hydra vulgaris and the marine colonial Hydractinia echinata; we have termed the Hydra gene HvSRF and the Hydractinia gene HeSRF. The MADS-box of both genes is identical in sequence and more similar to SRFs of other organisms than to non-SRF MADS-box-containing proteins from other organisms. Within the N terminus of the predicted protein, a motif of 14 amino acids is nearly identical between Hydra and Hydractinia. This motif is absent from other known SRF sequences. In the adult Hydra polyp, SRF is predominantly expressed in cells of the interstitial cell (I-cell) lineage. Expression of SRF ceases when I cells differentiate into nerve cells, nematocytes, or gland cells. In the course of sexual reproduction in Hydractinia, SRF is expressed in female germ cells. During embryogenesis, SRF transcripts are observed in all blastomeres. Later on, SRF expression is turned off in cells forming the ectodermal layer but further on is expressed in cells of the central cell mass, from which the endodermal epithelial cells and the I-cell lineage originate. Expression eventually becomes restricted to the I-cell lineage. We conclude that hydrozoan SRF is expressed in all these cells, which still have the property for differentiation. In adult Hydra, the abundance of SRF transcripts varies during the day. The pacemaker of this diurnal rhythm is the feeding regime. HvSRF expression decreases by 4 h after feeding and returns to the initial level 12 h after feeding. When feeding is stopped, the cycle of SRF expression persists through the first day when the animals are not fed. It has been shown that feeding partly synchronizes the cell cycle of the epithelial cells but not that of the I-cells. We suggest that the epithelial cells affect SRF expression in I-cells and thereby influence the decision of I-cells to enter a differentiation pathway. PMID- 11476574 TI - Hoxa3 and pax1 regulate epithelial cell death and proliferation during thymus and parathyroid organogenesis. AB - The thymus and parathyroid glands in mice develop from a thymus/parathyroid primordium that forms from the endoderm of the third pharyngeal pouch. We investigated the molecular mechanisms that promote this unique process in which two distinct organs form from a single primordium, using mice mutant for Hoxa3 and Pax1. Thymic ectopia in Hoxa3(+/-)Pax1(-/-) compound mutants is due to delayed separation of the thymus/parathyroid primordium from the pharynx. The primordium is hypoplastic at its formation, and has increased levels of apoptosis. The developing third pouch in Hoxa3(+/-)Pax1(-/-) compound mutants initiates normal expression of the parathyroid-specific Gcm2 and thymus-specific Foxn1 genes. However, Gcm2 expression is reduced at E11.5 in Pax1(-/-) single mutants, and further reduced or absent in Hoxa3(+/-)Pax1(-/-) compound mutants. Subsequent to organ-specific differentiation from the shared primordium, both the parathyroids and thymus developed defects. Parathyroids in compound mutants were smaller at their formation, and absent at later stages. Parathyroids were also reduced in Pax1(-/-) mutants, revealing a new function for Pax1 in parathyroid organogenesis. Thymic hypoplasia at later fetal stages in compound mutants was associated with increased death and decreased proliferation of thymic epithelial cells. Our results suggest that a Hoxa3-Pax1 genetic pathway is required for both epithelial cell growth and differentiation throughout thymus and parathyroid organogenesis. PMID- 11476575 TI - Endoderm specification and differentiation in Xenopus embryos. AB - It is known from work with amniote embryos that regional specification of the gut requires cell-cell signalling between the mesoderm and the endoderm. In recent years, much of the interest in Xenopus endoderm development has focused on events that occur before gastrulation and this work has led to a different model whereby regional specification of the endoderm is autonomous. In this paper, we examine the specification and differentiation of the endoderm in Xenopus using neurula and tail-bud-stage embryos and we show that the current hypothesis of stable autonomous regional specification is not correct. When the endoderm is isolated alone from neurula and tail bud stages, it remains fully viable but will not express markers of regional specification or differentiation. If mesoderm is present, regional markers are expressed. If recombinations are made between mesoderm and endoderm, then the endodermal markers expressed have the regional character of the mesoderm. Previous results with vegetal explants had shown that endodermal differentiation occurs cell-autonomously, in the absence of mesoderm. We have repeated these experiments and have found that the explants do in fact show some expression of mesoderm markers associated with lateral plate derivatives. We believe that the formation of mesoderm cells by the vegetal explants accounts for the apparent autonomous development of the endoderm. Since the fate map of the Xenopus gut shows that the mesoderm and endoderm of each level do not come together until tail bud stages, we conclude that stable regional specification of the endoderm must occur quite late, and as a result of inductive signals from the mesoderm. PMID- 11476576 TI - Sox3 expression defines a common primordium for the epibranchial placodes in chick. AB - The epibranchial placodes are ectodermal thickenings that generate sensory neurons of the distal ganglia of the branchial nerves. Although significant advances in our understanding of neurogenesis from the placodes have recently been made, the events prior to the onset of neurogenesis remain unclear. We found that chick Sox3 (cSox3) shows a highly dynamic pattern of expression before becoming confined to the final placodes: one pre-otic (geniculate) and three post otic (one petrosal and two nodose) placodes. A fate-mapping study using lipophilic dyes revealed that all post-otic placodes arise within a single broad cSox3-positive domain, where cSox3 expression and epithelial thickness will be retained only in much smaller final neurogenic placodes. The data presented here suggest that post-otic placodes are remnants of a common primordium defined as a discrete domain of cSox3 expression. PMID- 11476577 TI - A population of caudally migrating cranial neural crest cells: functional and evolutionary implications. AB - The deployment of the cranial neural crest is central to the patterning of the skeletomuscular elements of the vertebrate head, with cranial muscles invariably attaching to skeletal elements formed by crest from the same axial level. Here we demonstrate, through gene expression analysis, ablation studies and fate-mapping, the existence of a population of caudally migrating cranial crest that arise from the postotic neural tube. As with the rest of the postotic crest, these cells express the transcription factor Mafb, and this marker can be used to highlight their posterior migration. They pass out between the anterior somite and the otic vesicle, before turning caudally and running along the base of the somites. With long-term fate mapping, we show that these cells migrate to the clavicle and settle at the site of formation of the attachment point for the cleidohyoid muscle. As such, the influence of the cranial neural crest in organising skeletomuscular connectivity seems to extend beyond the head into the trunk. These results are of further importance as they help explain how, even though the pectoral girdle and the skull became physically dissociated during tetrapod evolution, skeletomuscular connectivity has been maintained. PMID- 11476578 TI - The whereabouts of a morphogen: direct evidence for short- and graded long-range activity of hedgehog signaling peptides. AB - Sonic Hedgehog (Shh) and Indian Hedgehog (Ihh) are members of the Hedgehog (Hh) family of signaling molecules known to be involved in embryonic patterning and morphogenesis. The Hh proteins undergo an autocatalytic cleavage to yield an N terminal and a C-terminal peptide, with the signaling capacities confined to the N peptide. Drosophila Hh-N has been shown to act via both short- and long-range signaling. In vertebrates, however, attempts to directly demonstrate Shh (SHH) or Ihh (IHH) proteins at a distance from producing cells have been largely unsuccessful. Furthermore, the fact that the Hh N peptides occur in a cholesterol modified, membrane-tethered form is not easily reconciled with long-range signaling. This study used optimized immunohistochemistry combined with tissue separation and biochemical analyses in vivo and in vitro to determine the range of action of SHH and IHH in the mouse embryo. In all embryonic structures studied, we detect signaling peptides in producing cells, but we also find that ligands move over considerable distances depending on the tissue. These data provide direct evidence for the presence of Hedgehog signaling peptides in target compartments, suggesting a direct long-range action without a need for secondary mediators. Visualization of Hedgehog proteins in target tissues was achieved only under conditions that allowed proteoglycan/glycosaminoglycan (PG/GAG) preservation. Furthermore, we show that induced changes of the composition of PG/GAG in the tooth alter SHH signaling. These data suggest a crucial role for PG/GAGs in Hedgehog movement. PMID- 11476579 TI - Imprinted expression of neuronatin from modified BAC transgenes reveals regulation by distinct and distant enhancers. AB - Neuronatin (Nnat) is an imprinted gene that is expressed exclusively from the paternal allele while the maternal allele is silent and methylated. The Nnat locus exhibits some unique features compared with other imprinted domains. Unlike the majority of imprinted genes, which are organised in clusters and coordinately regulated, Nnat does not appear to be closely linked to other imprinted genes. Also unusually, Nnat is located within an 8-kb intron of the Bc10 gene, which generates a biallelically expressed, antisense transcript. A similar organisation is conserved at the human NNAT locus on chromosome 20. Nnat expression is first detected at E8.5 in rhombomeres 3 and 5, and subsequently, expression is widespread within postmitotic neuronal tissues. Using modified BAC transgenes, we show that imprinted expression of Nnat at ectopic sites requires, at most, an 80 kb region around the gene. Furthermore, reporter transgenes reveal distinct and dispersed cis-regulatory elements that direct tissue-specific expression and these are predominantly upstream of the region that confers allele-specific expression. PMID- 11476580 TI - CLIP-50 immunolocalization during mouse spermiogenesis suggests a role in shaping the sperm nucleus. AB - The spermatid nucleus and cytoplasm undergo dramatic morphological modifications during spermatid differentiation into mature sperm. Some of the external force causing this nuclear shaping is generated by a microtubular structure termed the manchette, which attaches to the perinuclear ring of the spermatid. Here, we report the isolation and characterization of a protein component of this perinuclear ring in an immunological screening of a mouse testis cDNA library. We termed this protein CLIP-50 because of its high similarity at the amino acid level to the C-terminal region of the microtubule-binding protein CLIP 170/restin. CLIP-50 lacks the characteristic microtubule-binding motif, but retains a portion of the predicted coiled-coiled domain and the metal-binding motif. The CLIP-50 transcript and protein are abundant in testis. The protein is also expressed in heart, lung, kidney, and skin. A distinct size variant exists in brain. In the spermatids, CLIP-50 protein localizes specifically to the centriolar region where the sperm tail originates and to the perinuclear ring from which the manchette emerges. CLIP-50 staining is retained in the ring throughout its migration over the surface of the nucleus which accompanies the nuclear shaping into its characteristic sperm configuration. This localization pattern indicates a very specific function for this novel CLIP derivative during mouse spermiogenesis. PMID- 11476581 TI - Genetic dissection of the Drosophila Cubitus interruptus signaling complex. AB - Much of our understanding of the Hedgehog (Hh) signaling pathway comes from Drosophila, where a gradient of Hh signaling regulates the function of the transcription factor Cubitus interruptus (Ci) at three levels: protein stabilization, nuclear import, and activation. Regulation of Ci occurs in a cytoplasmic complex containing Ci, the kinesin-like protein Costal-2 (Cos2), the serine-threonine kinase Fused (Fu), and the Suppressor of Fused [Su(fu)] protein. The mechanisms by which this complex responds to different levels of Hh signaling and establishes distinct domains of gene expression are not fully understood. By sequentially mutating components from the Ci signaling complex, their roles in each aspect of Ci regulation can be analyzed. The Cos2-Ci core complex is able to mediate Hh-regulated activation of Ci but is insufficient to regulate nuclear import and cleavage. Addition of Su(fu) to the core complex blocks nuclear import while the addition of Fu restores Hh regulation of Ci nuclear import and proteolytic cleavage. Fu participates in two partially redundant pathways to regulate Ci nuclear import; the kinase function plays a positive role by inhibiting Su(fu), and the regulatory domain plays a negative role in conjunction with Cos2. PMID- 11476582 TI - Manifestation of the limb prepattern: limb development in the absence of sonic hedgehog function. AB - The secreted protein encoded by the Sonic hedgehog (Shh) gene is localized to the posterior margin of vertebrate limb buds and is thought to be a key signal in establishing anterior-posterior limb polarity. In the Shh(-/-) mutant mouse, the development of many embryonic structures, including the limb, is severely compromised. In this study, we report the analysis of Shh(-/-) mutant limbs in detail. Each mutant embryo has four limbs with recognizable humerus/femur bones that have anterior-posterior polarity. Distal to the elbow/knee joints, skeletal elements representing the zeugopod form but lack identifiable anterior-posterior polarity. Therefore, Shh specifically becomes necessary for normal limb development at or just distal to the stylopod/zeugopod junction (elbow/knee joints) during mouse limb development. The forelimb autopod is represented by a single distal cartilage element, while the hindlimb autopod is invariably composed of a single digit with well-formed interphalangeal joints and a dorsal nail bed at the terminal phalanx. Analysis of GDF5 and Hoxd11-13 expression in the hindlimb autopod suggests that the forming digit has a digit-one identity. This finding is corroborated by the formation of only two phalangeal elements which are unique to digit one on the foot. The apical ectodermal ridge (AER) is induced in the Shh(-/-) mutant buds with relatively normal morphology. We report that the architecture of the Shh(-/-) AER is gradually disrupted over developmental time in parallel with a reduction of Fgf8 expression in the ridge. Concomitantly, abnormal cell death in the Shh(-/-) limb bud occurs in the anterior mesenchyme of both fore- and hindlimb. It is notable that the AER changes and mesodermal cell death occur earlier in the Shh(-/-) forelimb than the hindlimb bud. This provides an explanation for the hindlimb-specific competence to form autopodial structures in the mutant. Finally, unlike the wild-type mouse limb bud, the Shh(-/-) mutant posterior limb bud mesoderm does not cause digit duplications when grafted to the anterior border of chick limb buds, and therefore lacks polarizing activity. We propose that a prepattern exists in the limb field for the three axes of the emerging limb bud as well as specific limb skeletal elements. According to this model, the limb bud signaling centers, including the zone of polarizing activity (ZPA) acting through Shh, are required to elaborate upon the axial information provided by the native limb field prepattern. PMID- 11476583 TI - The perplexed and confused mutations affect distinct stages during the transition from proliferating to post-mitotic cells within the zebrafish retina. AB - To identify and study genes essential for vertebrate retinal development, we are screening zebrafish embryos for mutations that disrupt retinal histogenesis. Key steps in retinogenesis include withdrawal from mitosis by multipotent neuroepithelial cells, specification to particular cell types, migration to the appropriate laminar positions, and molecular and morphological differentiation. In this study, we have identified two recessive mutations that affect the transition of proliferating neuroepithelial cells to postmitotic retinal cells. Both the perplexed and confused mutant phenotypes were initially detectable when the first retinal neuroepithelial cells began to leave the cell cycle. At this time, each mutant retina showed increased cell death and a lack of morphological differentiation. Cell death was found to be apoptotic in both perplexed and confused retinas based on TUNEL analysis and activation of caspase-3. TUNEL phosphoRb-BrdU colocalization studies indicated that the perplexed mutation caused death in cells transitioning from a proliferative to postmitotic state. For the confused mutation, TUNEL-phosphoRb-BrdU analysis revealed that only a subset of postmitotic cells were induced to activate apoptosis. Mosaic analysis demonstrated that within the retina the perplexed mutation functions noncell autonomously. Furthermore, whole lens or eye cup transplantations indicated that the retinal defect was intrinsic to the retina. Mosaic analysis with confused embryos showed this mutation acts cell-autonomously. From these studies, we conclude that the perplexed and confused genes are essential at distinct stages during the transition from proliferating to postmitotic cells within the zebrafish retina. PMID- 11476584 TI - Microenvironmental regulation of visual pigment expression in the chick retina. AB - Visual pigment (VP) expression in the chick embryo retina was investigated in ovo, in dissociated and explant cultures, and in cDNAs from individual cells. While VP mRNA is not detectable by in situ hybridization until embryonic day (ED) 14-16 in ovo, analysis of VP expression by RT-PCR showed that VP messages are present in the retina as many as 7-10 days before they become detectable by in situ hybridization, and are also detected in other regions of the embryonic CNS. On the other hand, red opsin expression is markedly accelerated when cells are isolated from their intraocular microenvironment at ED 6, and placed in pigment epithelium-free dissociated or explant cultures. This acceleration occurs regardless of cell density, birth date, or serum presence in the medium, suggesting that many photoreceptors are already programmed to express red opsin on or before ED 6, and that microenvironmental inhibitory factors prevent implementation of this program until ED 14 in ovo. The selectivity of this phenomenon is suggested by the finding that other VPs are not observed by in situ hybridization in ED 6 cultures, although they are detectable in cultures of older retinas. Taken together, these findings suggest that red opsin expression may be constitutive for many developing photoreceptor cells in the chick. PMID- 11476585 TI - Boundaries and functional domains in the animal/vegetal axis of Xenopus gastrula mesoderm. AB - Patterning of the Xenopus gastrula marginal zone in the axis running equatorially from the Spemann organizer-the so--called "dorsal/ventral axis"--has been extensively studied. It is now evident that patterning in the animal/vegetal axis also needs to be taken into consideration. We have shown that an animal/vegetal pattern is apparent in the marginal zone by midgastrulation in the polarized expression domains of Xenopus brachyury (Xbra) and Xenopus nodal-related factor 2 (Xnr2). In this report, we have followed cells expressing Xbra in the presumptive trunk and tail at the gastrula stage, and find that they fate to presumptive somite, but not to ventrolateral mesoderm of the tailbud embryo. From this, we speculate that the boundary between the Xbra- and Xnr2-expressing cells at gastrula corresponds to a future tissue boundary. In further experiments, we show that the level of mitogen-activated protein kinase (MAPK) activation is polarized along the animal/vegetal axis, with the Xnr2-expressing cells in the vegetal marginal zone having no detectable activated MAPK. We show that inhibition of MAPK activation in Xenopus animal caps results in the conversion of Xnr2 from a dorsal mesoderm inducer to a ventral mesoderm inducer, supporting a role for Xnr2 in induction of ventral mesoderm. PMID- 11476586 TI - Lilliputian mutant of maize lacks cell elongation and shows defects in organization of actin cytoskeleton. AB - The maize mutant lilliputian is characterized by miniature seedling stature, reduced cell elongation, and aberrant root anatomy. Here, we document that root cells of this mutant show several defects in the organization of actin filaments (AFs). Specifically, cells within the meristem lack dense perinuclear AF baskets and fail to redistribute AFs during mitosis. In contrast, mitotic cells of wild type roots accumulate AFs at plasma membrane-associated domains that face the mitotic spindle poles. Both mitotic and early postmitotic mutant cells fail to assemble transverse arrays of cortical AFs, which are characteristic for wild type root cells. In addition, early postmitotic cells show aberrant distribution of endoplasmic AF bundles that are normally organized through anchorage sites at cross-walls and nuclear surfaces. In wild-type root apices, these latter AF bundles are organized in the form of symmetrically arranged conical arrays and appear to be essential for the onset of rapid cell elongation. Exposure of wild type and cv. Alarik maize root apices to the F-actin drugs cytochalasin D and latrunculin B mimics the phenotype of lilliputian root apices. In contrast to AFs, microtubules are more or less normally organized in root cells of lilliputian mutant. Collectively, these data suggest that the LILLIPUTIAN protein, the nature of which is still unknown, impinges on plant development via its action on the actin cytoskeleton. PMID- 11476587 TI - Evidence that phospholipase C from the sperm is not responsible for initiating Ca(2+) release at fertilization in mouse eggs. AB - Release of Ca(2+) from intracellular stores at fertilization of mammalian eggs is mediated by inositol 1,4,5-trisphosphate (IP3), but the mechanism by which the sperm initiates IP3 production is not yet understood. We tested the hypothesis that phospholipase C (PLC) activity introduced into the mouse egg as a consequence of sperm-egg fusion is responsible for causing Ca(2+) release. We demonstrated that microinjecting purified, recombinant PLCgamma1 protein into mouse eggs caused Ca(2+) oscillations like those seen at fertilization. However, the PLC activity in the minimum amount of purified PLCgamma1 protein needed to elicit Ca(2+) release when injected into eggs was approximately 500-900 times the PLC activity contained in a single sperm. This indicates that a single mouse sperm does not contain enough PLC activity to be responsible for causing Ca(2+) release at fertilization. We also examined whether phosphatidylinositol 3-kinase (PI3K) could have a role in this process, and found that several inhibitors of PI3K-mediated signaling had no effect on Ca(2+) release at fertilization. PMID- 11476588 TI - Guinea pig fertilin exhibits restricted lateral mobility in epididymal sperm and becomes freely diffusing during capacitation. AB - The guinea pig sperm protein fertilin functions in sperm-egg plasma membrane binding. Fertilin is initially present in the plasma membrane of the whole head in testicular sperm, then becomes concentrated into the posterior head domain during epididymal passage. Fertilin remains localized to the posterior head plasma membrane following the acrosome reaction, when it functions in sperm-egg interaction. Fluorescence redistribution after photobleaching was used to examine the lateral mobility of fertilin in both acrosome-intact and acrosome-reacted sperm. Fertilin exhibited highly restricted lateral mobility in both testicular and epididymal sperm (D < 10(-10) cm(2)/s). However, fertilin in acrosome-reacted sperm was highly mobile within the membrane bilayer (D = 1.8 x 10(-9) cm(2)/s and %R = 84). Measurement of the lateral mobility of fertilin in capacitated, acrosome-intact sperm revealed two populations of cells. In approximately one half of the cells, lateral mobility of fertilin was similar to sperm freshly isolated from the cauda epididymis; while in the other half fertilin was highly mobile. The release of fertilin from interactions that restrict its lateral mobility may regulate its function in sperm-egg interaction. PMID- 11476589 TI - Neuropathology of mice carrying mutant APP(swe) and/or PS1(M146L) transgenes: alterations in the p75(NTR) cholinergic basal forebrain septohippocampal pathway. AB - Cholinergic basal forebrain (CBF) projection systems are defective in late Alzheimer's disease (AD). We examined the brains of 12-month-old singly and doubly transgenic mice overexpressing mutant amyloid precursor protein (APP(swe)) and/or presenilin-1 (PS1(M146L)) to investigate the effects of these AD-related genes on plaque and tangle pathology, astrocytic expression, and the CBF projection system. Two types of beta-amyloid (Abeta)-immunoreactive (ir) plaques were observed: type 1 were darkly stained oval and elongated deposits of Abeta, and type 2 were diffuse plaques containing amyloid fibrils. APP(swe) and PS1(M146L) mouse brains contained some type 1 plaques, while the doubly transgenic (APP(swe)/PS1(M146L)) mice displayed a greater abundance of types 1 and 2 plaques. Sections immunostained for the p75 NGF receptor (p75(NTR)) revealed circular patches scattered throughout the cortex and hippocampus of the APP(swe)/PS1(M146L) mice that contained Abeta, were innervated by p75(NTR)-ir neurites, but displayed virtually no immunopositive neurons. Tau pathology was not seen in any transgenic genotype, although a massive glial response occurred in the APP(swe)/PS1(M146L) mice associated with amyloid plaques. Stereology revealed a significant increase in p75(NTR)-ir medial septal neurons in the APP(swe) and PS1(M146L) singly transgenic mice compared to the APP(swe)/PS1(M146L) mice. No differences in size or optical density of p75(NTR) ir neurons were observed in these three mutants. p75(NTR)-ir fibers in hippocampus and cortex were more pronounced in the APP(swe) and PS1(M146L) mice, while the APP(swe)/PS1(M146L) mice showed the least p75(NTR)-ir fiber staining. These findings suggest a neurotrophic role for mutant APP and PS1 upon cholinergic hippocampal projection neurons at 12 months of age. PMID- 11476590 TI - Changes in metabotropic glutamate receptor expression following spinal cord injury. AB - Spinal cord injury (SCI) initiates biochemical events that lead to an increase in extracellular excitatory amino acid concentrations, resulting in glutamate receptor-mediated excitotoxic events. These receptors include the three groups of metabotropic glutamate receptors (mGluRs). Group I mGluR activation can initiate a number of intracellular pathways that increase neuronal excitability. Group II and III mGluRs may function as autoreceptors to modulate neurotransmission. Thus, all three groups may contribute to the mechanisms of central sensitization and chronic central pain. To begin evaluating mGluRs in SCI, we quantified the changes in mGluR expression after SCI in control (naive), sham, and impact injured adult male Sprague-Dawley rats (200-250 g). SCI was produced at spinal segment T10 with a New York University impactor (12.5-mm drop, 10-g rod of 2-mm diameter). Expression levels were determined by Western blot and immunohistochemistry analyses at the epicenter of injury, as well as segments rostral and caudal. The group I subtype mGluR1 was increased over control levels in segments rostral and caudal by postsurgical day (PSD) 7 and remained elevated through PSD 60. The group I subtype mGluR5 was unchanged in all segments rostral and caudal to the injury at every time point measured. Group II mGluRs were decreased compared to control levels from PSD 7 through PSD 60 in all segments. These results suggest that different subtypes of mGluRs have different spatial and temporal expression patterns following SCI. The expression changes in mGluRs parallel the development of mechanical allodynia and thermal hyperalgesia following SCI; therefore, understanding the expression of mGluRs after SCI may give insight into mechanisms underlying the development of chronic central pain. PMID- 11476591 TI - Neuroprotection of grafted neurons with a GDNF/caspase inhibitor cocktail. AB - Transplantation of fetal ventral mesencephalic (VM) tissue shows great promise as an experimental therapy for patients with Parkinson's disease. However, cell survival in brain tissue grafts is poor, with survival rates of only 5-15%. We have utilized a combination of the caspase inhibitor bocaspartyl (OMe) fluoromethylketone (BOC-ASP-CH2F) and glial cell line-derived neurotrophic factor (GDNF) to enhance survival of grafted dopamine neurons. The VM tissue was dissected from embryonic day 13-15 rat fetuses, incubated in different doses of BOC-ASP-CH2F and GDNF, and transplanted to the anterior chamber of the eye of adult rats. Growth of the tissue was assessed through the translucent cornea. Doses of 50 and 100 micromolar of the general caspase inhibitor appeared to have detrimental effects on mesencephalic tissue, while 20 micromolar had beneficial effects on overall transplant growth. A combination of the caspase inhibitor and GDNF appeared to have more prominent effects on cell survival as well as dopaminergic fiber density than either agent by itself. The transplants doubled in size when they were treated with a combination of BOC-ASP-CH2F and GDNF, and cell death markers were significantly reduced at both 48 h and 4-6 days postgrafting. This is, to our knowledge, the first combined approach using apoptotic blockers with trophic factors, and demonstrates a viable strategy for protection of developing neurons, since several different aspects of graft function may be addressed simultaneously. PMID- 11476592 TI - Amyloid beta-peptide inhibits neuronal glucose uptake by preventing exocytosis. AB - Amyloid beta peptide (Abeta) is suspected as a contributing factor for decreased glucose utilization in the brain of Alzheimer's patients; however, little is known about the regulatory mechanism of neuronal glucose uptake and how Abeta affects such a mechanism. We report that membrane depolarization by 40 mM KCl increases both neuronal glucose uptake and immunolabeling of the exofacial epitope of glucose transporter isoform GLUT3, suggesting that fusion of GLUT3 vesicles with the plasma membrane increases glucose uptake. Abeta25-35 decreased neuronal glucose uptake and this decrease was prevented by exocytosis-enhancing compounds (40 mM KCl, 50 microM ruthenium red). Abeta25-35 also inhibited exocytosis of the fluorescent membrane dye FM1-43 at neuronal cell bodies; however, 40 mM KCl was effective in overcoming this Abeta inhibition. Furthermore, GLUT3 colocalized with SNARE (N-ethylmaleimide-sensitive factor attached protein receptor) complex proteins (SNAP-25 and Syntaxin 1), and cleavage of the v-SNARE, VAMP, reduced glucose uptake. Our findings suggest that neuronal glucose uptake is regulated by SNARE complex-dependent docking and fusion of GLUT3 vesicles with the plasma membrane and that Abeta decreases glucose uptake by inhibiting fusion of these vesicles. PMID- 11476593 TI - Proliferation and regeneration of retrogradely labeled adult rat corticospinal neurons in culture. AB - These are the first studies to demonstrate that adult rat corticospinal tract (CST) neurons, which were identified by retrograde neuronal labeling, retain regenerative and proliferative potential. To determine if adult CST neurons undergo cell division, we tested if these retrogradely labeled cells synthesize DNA by adding BrdU to the cultures 24 h prior to fixation of the cells. The result shows that adult corticospinal tract neurons are capable of DNA synthesis, and our total cell counts with labeled cells counts further suggest that these cells undergo cell division. PMID- 11476594 TI - Time course study of GFRalpha-1 expression in an animal model of stroke. AB - Previous studies have shown that intracerebral administration of glial cell line derived neurotrophic factor (GDNF) reduces ischemia-mediated cerebral infarction. The biological effects of GDNF are mediated by GDNF-family receptor alpha-1 (GFRalpha-1) and c-Ret. In this study, we examined the levels of expression of GFRalpha-1 and c-Ret in a rat model of stroke. Adult Sprague-Dawley rats were anesthetized with chloral hydrate. The right middle cerebral artery was ligated at its distal branch for 90 min. Animals were sacrificed at 0, 6, 12, and 24 h after reperfusion and levels of expression of GFRalpha-1 and c-Ret mRNA were determined by in situ hybridization histochemistry. We found that GFRalpha-1 mRNA was up-regulated in CA3, dentate gyrus (DG), cortex, and striatum. The peak of up regulation in DG was 6 h after reperfusion. GFRalpha-1 mRNA levels in CA3 were gradually up-regulated over the 24-h reperfusion period. In cortex, GFRalpha-1 mRNA was up-regulated at all time points; however, the peak of up-regulation was observed at 0 and 24 h after reperfusion. In striatum, an initial up-regulation of GFRalpha-1 was found at 0 h after ischemia. In striatum, up-regulation of c Ret mRNA was detected as early as 0 h after reperfusion. A gradual increase was found at 6, 12, and 24 h after reperfusion. In conclusion, our results indicate that there are both regional and temporal differences in up-regulation of GFRalpha-1 and c-Ret after ischemia. Since GDNF is neuroprotective, up-regulation of GFRalpha-1 and c-Ret could enhance the responsiveness to GDNF and reduce neuronal damage. The selective up-regulation of GFRalpha-1 and c-Ret in different brain areas suggests that there may be regional differences in GDNF-induced neuroprotection in stroke. PMID- 11476595 TI - Cutaneous nerve terminal degeneration in painful mononeuropathy. AB - Nociceptive nerves innervate the skin and play an important role in the generation of neuropathic pain. However, it remains elusive whether and how nociceptive nerve terminals degenerate in neuropathic pain conditions. To address this issue, we investigated cutaneous innervation in a model of painful mononeuropathy, the chronic constriction injury (CCI). The hind paws of rats were immunocytochemically stained with a pan-axonal marker, protein gene product 9.5 (PGP 9.5). Within 2 days after CCI, rats exhibited thermal hyperalgesia, and there was a partial depletion of epidermal nerves. The extent of reduction in epidermal nerves after CCI was variable with an epidermal nerve density of 3.65 +/- 1.97 fibers/mm (compared to 15.39 +/- 1.58 fibers/mm on the control side, P < 0.02). There was a mild but concomitant increase in PGP 9.5 (+) Langerhans cells in the epidermis of the skin with CCI (10.19 +/- 1.99 vs 7.75 +/- 1.36 cells/mm, P < 0.05). In the skin denervated by tight ligation of the sciatic nerve, epidermal nerves were completely depleted (0 fibers/mm vs. 12.26 +/- 1.44 fibers/mm on the control side, P < 0.001). Animals with tight ligation of the sciatic nerve exhibited thermal anesthesia. These findings suggest that the epidermis is partially denervated in CCI, and that a partial injury of nerves is correlated with the development of neuropathic pain. PMID- 11476596 TI - Effective reduction of neuronal death by inhibiting gap junctional intercellular communication in a rodent model of global transient cerebral ischemia. AB - Gap junctions assemble astrocytes into syncytia, allowing exchange of metabolites, catabolites, and second-messenger molecules. Connexin43 is the predominant connexin of astrocytic gap junctions. The distribution of gap junction protein connexin43 was analyzed in different subfields of the hippocampal formation as a function of time after transient forebrain ischemia. One decisive key step in understanding why an ischemic insult gradually expands may be to establish how gap junction channels permit dying cells in the ischemic focus to communicate, in particular, with viable cells. The role of gap junctional intercellular communication in the hippocampus under ischemic conditions could be decisive for cell death propagation. We found that the vulnerable CA1/CA2 subfields have a higher density of gap junctions than the resistant CA3/CA4 areas, that changes in the distribution of connexin43 immunoreactivity may correlate with the phenomenon of selective vulnerability, and that inhibition of astrocytic gap junction permeability by octanol restricts the flow of undesirable neurotoxins that could potentially exacerbate neuronal damage. This provides a novel perspective for analysis of the pathophysiology of cerebral ischemia. PMID- 11476597 TI - Solid human embryonic spinal cord xenografts in acute and chronic spinal cord cavities: a morphological and functional study. AB - While therapeutic spinal cord grafting procedures are of interest in the chronic spinal cord injury stage, previous experimental grafting studies, including human spinal cord tissue, have mainly focused on the acute stage. Therefore, solid human embryonic spinal cord grafts were implanted in acute or chronic spinal cord aspiration cavities of immunodeficient rats to compare the morphological and locomotor outcome to that of lesion alone cases. Locomotor function was assessed using the Basso, Beattie, and Bresnahan open-field locomotor rating scale up to 6 months, while the morphological evaluation of graft survival, growth, and integration was performed at 6 weeks or 6 months after implantation. Graft survival was 94% in both lesion models, while graft growth was enhanced in the chronic compared to the acute cavity group. Human specific Thy-1 and neurofilament immunoreactive fibers were observed up to 7 mm into host white matter, while aminergic fibers were observed up to 1 mm into the grafts. Abundant calcitonin gene-related peptide immunoreactive fibers in the grafts in the absence both of immunoreactive cell bodies and colocalized human-specific neurofilament immunoreactivity, suggested host fiber ingrowth. At 6 months, the grafted cases presented less central canal deformation and lower glial fibrillary acidic protein immunoreactivity at the host cavity border compared to that of the nongrafted cases. The strong compensatory regain of locomotor function after unilateral spinal cord lesions was not affected by the human spinal cord grafts. In conclusion, solid human embryonic spinal cord tissue transplanted to a cavity in the adult injured spinal cord results in beneficial morphological effects in both the acute and chronic spinal cord lesion. PMID- 11476598 TI - Long-term proliferation and dopaminergic differentiation of human mesencephalic neural precursor cells. AB - We report on generation of dopamine neurons from long-term cultures of human fetal mesencephalic precursor cells. These CNS precursor cells were successfully expanded in vitro using the mitogens epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2). Incubation of these cultures in 3% atmospheric oxygen resulted in higher cellular yields than room air. Following incubation in differentiation media containing interleukin (IL)-1b (IL-1b), IL-11, leukemia inhibitory factor (LIF), and glial cell line-derived neurotrophic factor (GDNF), up to 1% of the precursor cells converted into cells immunoreactive for tyrosine hydroxylase (TH), a marker for dopamine neurons. The TH immunoreactive cells exhibited morphological and functional properties characteristic of dopamine neurons in culture. These precursor cells might serve as a useful source of human dopamine neurons for studying the development and degeneration of human dopamine neurons and may further serve as a continuous, on-demand source of cells for therapeutic transplantation in patients with Parkinson's disease. PMID- 11476599 TI - Behavioral phenotyping of GFAP-apoE3 and -apoE4 transgenic mice: apoE4 mice show profound working memory impairments in the absence of Alzheimer's-like neuropathology. AB - For the purpose of studying the potential neurobehavioral effects of different human apolipoprotein E (apoE) isoforms produced within the brain, transgenic (TG) mice were generated in which human apoE3 or apoE4 isoforms were under control of an astrocyte-specific, glial fibrillary acidic protein promoter and these TG mice were bred back to apoE knockout (KO) mice. Behavioral phenotypes of apoE3 and apoE4 TG mice were derived by conducting a longitudinal study in which apoE3 and apoE4 TG mice were compared with apoE KO and wild-type (WT) mice (all male) on several behavioral measures. Analysis of locomotor activity, "open-field" behaviors, acoustic startle/prepulse inhibition, and elevated plus maze data suggested that the apoE TG/KO groups were more "emotionally reactive" than WT mice, with apoE4 mice typically being the most reactive. The absence of performance differences among groups on the rotating holeboard and water navigation tasks suggested intact reference memory processing in apoE TG/KO mice. However, apoE4 mice were profoundly impaired on a working memory-based protocol in the radial arm maze (11-14 months). Nonassociative factors (sensorimotor capacities or emotionality differences) did not appear to confound interpretation of the learning/memory results. Western blot analysis revealed no alterations in the level of synaptic, neuronal, or glial markers in neocortex or hippocampus and histologic analysis revealed no evidence of Abeta deposition or neuritic plaques in the apoE KO/TG mice. Our findings suggest that apoE4 expression in the brain may have selective deleterious effects on memory function in the absence of typical Alzheimer's-like neuropathology. PMID- 11476600 TI - Length of postovariectomy interval and age, but not estrogen replacement, regulate N-methyl-D-aspartate receptor mRNA levels in the hippocampus of female rats. AB - Estrogens and N-methyl-D-aspartate (NMDA) receptors regulate multiple aspects of morphological and functional plasticity in young animals. For example, estrogens increase spine density in the hippocampus, and NMDA antagonists block these effects. Few studies have examined the effects of age, postovariectomy interval, and duration of estrogen replacement in the hippocampus and more specifically on NMDA receptor subunits. Therefore, the present study was designed to investigate the effects of short- and long-term estrogen replacement or deprivation on mRNA levels of three NMDA receptor subunits, NR1, NR2A, and NR2B, in the hippocampus of aging female Sprague-Dawley rats. Young (3- to 4-month-old) and middle-aged (12- to 13-month-old) rats were ovariectomized for 1 month and then treated with estrogen or vehicle for either 2 days or 2 weeks. Another set of middle-aged and aged (24-to 25-month-old) animals were ovariectomized for 6 months and treated with estrogen or vehicle for 2 days or 2 weeks. RNase protection assay was used to assess changes in the NMDA receptor subunit mRNA levels. Our results demonstrated significant effects of age and length of ovariectomy on NMDA receptor mRNA levels, with little effect of the estrogen status of the animals on these parameters. The largest effect was seen for the length of the postovariectomy interval, with the results demonstrating that rats with a short term ovariectomy have substantially higher NMDA receptor subunit mRNA levels than animals with long-term ovariectomy. The most dramatic effects of aging were seen for NR1 and NR2B mRNAs in ventral hippocampus, with large age-related increases. These data suggest that age and duration of ovariectomy impact NMDA receptor mRNA levels in the hippocampus, potentially affecting the stoichiometry and/or function of these receptors. These findings have important implications for postmenopausal or hysterectomy/oophorectomy estrogen depletion and replacement in humans. PMID- 11476601 TI - Increase in TNFalpha transport after SCI is specific for time, region, and type of lesion. AB - The dynamic changes of the blood-brain barrier and blood-spinal cord barrier (BBB) are an important part of the CNS response to injury. This study addresses the permeability of the BBB in the acute phase of spinal cord injury (SCI) to the thoracic region. SCI by compression or by complete transection was generated in mice. BBB disruption was evaluated by spinal cord uptake of radiolabeled albumin. The BBB of the thoracic spinal cord was disrupted immediately after compression injury, lasting for 2 days. This was followed by a delayed permeability increase in the cervical spinal cord beginning 3 days after injury. After transection, BBB disruption was limited to the thoracic spinal cord and was present only immediately postinjury. The entry of TNFalpha not only was increased at the time of BBB disruption, following the same pattern, but also had secondary changes after the BBB permeability to albumin had returned to normal. The increase of TNFalpha entry, best explained by upregulation of the specific transport system for TNFalpha, was pronounced in the lumbar spinal cord as well as the thoracic region, and followed a different time course after the two types of injury. Integrating our results with those of the literature regarding the roles of inflammatory responses and the effects of TNFalpha in spinal cord regeneration, we conclude that the time-, region-, and lesion-specificity of the upregulation of TNFalpha transport is part of the regulatory changes at the BBB in response to SCI. PMID- 11476602 TI - Reinnervation of the superior colliculus delays down-regulation of ephrin A2 in neonatal rat. AB - Although the adult mammalian optic nerve does not regenerate following lesion, in the neonatal rat, retinal ganglion cell (RGC) axons retain the capacity to grow across lesion sites in the brain. Following a brachial lesion at postnatal day 2 (P2), some RGC axons, together with ingrowing cortico-tectal axons, cross the lesion to reinnervate the superior colliculus (SC). Here we use immunohistochemistry to examine expression of the guidance cue ephrin A2 following a brachial lesion. Normal animals show a steady decrease in ephrin A2 immunoreactivity between P5 and P31, with a low rostral to high caudal gradient being evident only at P5. By contrast, after brachial lesion, values are significantly elevated rostrally at P5 and caudally at P12; moreover, a steep rostro-caudal gradient is present at both ages. By P31 values fall to normal levels. Following unilateral enucleation at P2, levels are not significantly different from normal. Our results show that innervation but not denervation triggers increased ephrin A2 expression after a brachial lesion. PMID- 11476603 TI - The effect of interburst intervals on measures of hippocampal LTP in the freely moving adult male rat. AB - An important factor in the induction and maintenance of long-term potentiation (LTP) is the tetanization paradigm. This paper presents the changes associated with the induction and maintenance of hippocampal LTP in the freely moving adult male rat, subjected to three different tetanization paradigms. These results indicate that specific LTP measures including (1) synaptic activation, as measured by the slope of the dentate granule cell population excitatory postsynaptic potential, and (2) cellular response, as measured by the dentate population spike amplitude, evoked by single-pulse stimulation of the medial perforant pathway are dependent on the interburst interval of the bursting paradigm commonly used in LTP studies. PMID- 11476604 TI - Chronic intraventricular infusion of glial cell line-derived neurotrophic factor (GDNF) rescues some cerebellar Purkinje cells from heredodegeneration. AB - Cerebellar Purkinje cells degenerate in shaker mutant rats. Glia cell line derived neurotrophic factor (GDNF) was chronically infused intraventricularly in an attempt to rescue mutant Purkinje cells from dying. Four weeks of chronic GDNF infusion delayed the degeneration of many but not all Purkinje cells. Surviving Purkinje cells formed spatially related groups interrupted by other groups of degenerated Purkinje cells. There was a positive correlation in GDNF-supported Purkinje cell survival and persistence of normal motor behaviors. PMID- 11476605 TI - N-methyl-D-aspartate receptor blockade during development induces short-term but not long-term changes in c-Jun and parvalbumin expression in the rat cervical spinal cord. AB - During postnatal development, N-methyl-D-aspartate receptor (NMDA-R) expression progressively decreases in ventral and deep dorsal horns. This transient expression might play a role in activity-dependent development of segmental circuitry. NMDA-Rs were blocked unilaterally in the lower cervical spinal cord using Elvax implants that released the NMDA-R antagonist MK-801 maximally over a 2-week period from postnatal day 7 (P7) onward. At P14, the ratio of c-Jun immunoreactive motoneurons ipsilateral/contralateral to the implants was significantly increased and the ratio of parvalbumin immunoreactive neurons decreased, compared to control implants. However, at P84, MK-801-treated and control spinal cords appeared the same. Therefore, NMDA-R blockade during development only transiently altered expression of activity-dependent proteins in the spinal cord, unlike lesions to the developing motor cortex, which we have previously shown to have a permanent effect. PMID- 11476606 TI - Cognitive strategies employed in trying to arrange a first date. AB - This article deals with how adolescents go about making a date. It uses a funnelling approach to elicit information which is then studied in more detail in a series of further steps. Study 1 used a short survey to obtain data concerning subjects' (n=410) experience of dating. Thereafter, 47 of these subjects selected situations and actions relevant to their dating experience, from two specially prepared lists. Study 1 provided information concerning the relevance of the lists, their comprehensibility and the frequency with which subjects had encountered specific situations or actions. In Study 2, 287 subjects completed the survey, 73 were selected and given revised versions of the lists. After selecting situations and actions, they were interviewed concerning their choice. Subject's responses are discussed with respect to social cognitive development in adolescence. PMID- 11476607 TI - Affective and behavioral responses to friends who neglect their friends for dating partners: influences of gender, jealousy and perspective. AB - Adolescents in the ninth grade (n=75) responded to a hypothetical situation involving an individual who neglects their friend as a result of their transition to steady dating. Results indicated that 53 per cent of girls and 32 per cent of boys reported past personal exclusion by their best friend who they perceived to be preoccupied by a romantic partner. Only 21 per cent of girls and 19 per cent of boys reported that they had ignored their best friend in the past while being preoccupied by a romantic partner. There were significant differences in emotional reactions of individuals depending on their gender, level of characteristic jealousy, and the perspective taken in the jealousy triangle. Analyses of suggested behavioral responses revealed a significant difference in verbosity depending on emotional reactions, gender, and level of characteristic jealousy. In general, the strategies adolescents' recommended for handling this situation were constructive, active responses that convey a continuing commitment to the relationship. PMID- 11476608 TI - Coping with stress in different phases of romantic development. AB - The degree to which coping with stress in romantic relationships is related to relationship development was examined longitudinally in a sample of early adolescents (n=107). Repeated measures ANOVAs revealed a significant decrease in romantic stress over time, while at the same time active coping with romantic stress increased. Concurrently, the quality of romantic relationships changed towards greater intimacy and affection. The results provide some support for recent theories of a developmental sequence in romantic relationships, according to which relationships progress from a more casual initial stage towards a more committed affection phase. In addition, the study illustrates that the multiple functions romantic activity can serve at different stages of development are accompanied by various types of stressors. PMID- 11476609 TI - Diverse aspects of dating: associations with psychosocial functioning from early to middle adolescence. AB - Theories imply that some aspects of adolescent dating behavior will be associated with individuals' positive psychosocial functioning, while other aspects will be associated with problems. This study addressed associations between diverse aspects of dating at age 16 and: (1) individual and social functioning at age 12 and at age 16; and (2) change in psychosocial functioning from age 12 to age 16. Controlling for physical maturity, overinvolvement in dating at age 16 was associated with poorer psychosocial functioning in early and middle adolescence and also predicted declines in functioning between the two ages. Level of dating experience and quality of romantic relationships were associated with social adaptation at age 16, especially in the friendship and dating domains. PMID- 11476610 TI - Adolescent romantic relationships: a look from the future. AB - Employing a narrative approach, 40 young adult romantic couples were interviewed about their current relationships as well as about their relationship with a romantic partner that lasted for at least about 2 months when they were aged 15 or 16. When analysed, the transcribed interviews were found to portray adolescent and young adulthood romantic relationships in different terms. Adolescent romance was described more as a form of companionship and was valued for its experiences of totality and enthusiasm. Young adulthood romance, by contrast, was described more as a mature relationship characterized by trust, support, and stability. In addition, adolescent romance was described as less enjoyable, enduring more relationship problems. Males and females described differently their adolescent romance in relation to the quality of their current romantic relationship. Males, but not females, related a current more positive perception of the relationship to a more positive perception of adolescent romance. Adolescent romance is discussed within the framework of the developmental processes that characterize the development of romantic relationships. PMID- 11476611 TI - Characteristics of closeness in adolescent romantic relationships. AB - This investigation was designed to describe characteristics of closeness in the romantic relationships of early, mid and late adolescents, and to determine whether adolescent reports of relationship authority and reciprocity are linked to perceptions of interdependence, interaction frequency, activity diversity, influence, and relationship duration. Age was positively associated with interdependence, daily social interaction, weekly activity diversity, and reciprocity but not with influence, authority, or relationship duration; gender was unrelated to all characteristics of closeness. Authority and reciprocity were each positively associated with relationship influence. Authority moderated associations between reciprocity and several characteristics of closeness such that reciprocity was positively linked to interdependence, daily social interaction, and weekly activity diversity, but only in relationships characterized by low levels of authority. Neither reciprocity nor authority was associated with relationship duration. PMID- 11476612 TI - The interpersonal context of romantic autonomy in adolescence. AB - This study explored the interpersonal context of romantic autonomy in mid adolescence by examining within-relationship links with romantic intimacy and affiliation, links across relationships with mothers and friends, and individual differences due to age and gender. Two hundred and thirty adolescents in grades 9, 10 and 11 with a current romantic partner, completed self-report questionnaires assessing their relationships. The results indicated that romantic autonomy was positively linked to romantic intimacy and affiliation, although this link differed according to the duration of the romantic relationship. The results also indicated that romantic autonomy was higher among girls than boys. Finally, romantic autonomy was uniquely linked to intimacy and autonomy in both maternal and friend relationships. Overall, this study highlights the interconnection of intimacy and autonomy in romantic relationships and the interconnections across relationships. PMID- 11476613 TI - The capacity for romantic intimacy: exploring the contribution of best friend and marital and parental relationships. AB - This study examined, in a longitudinal design, the contributions of three different relationships, namely marital, parent-child and best friend, to the capacity for intimacy in romantic relationships of Israeli male adolescents, as well as the mediating role of socio-emotional capacities. Eighty-four 17-year-old adolescents and their parents filled out questionnaires concerning the quality of these relational contexts. Four years later the Intimacy Status Interview was administered to the adolescents at the conclusion of their mandatory military service to examine closeness, separateness, and commitment within their romantic relationships. Results showed that all relational contexts were related to capacity for intimacy (directly or indirectly), with higher relational qualities associated with better capability for intimacy. The marital relationship was associated with intimacy through its effect on the parent-child relationships. The effects of the parent-child relationships on the capacity for intimacy were mediated through the adolescents' socio-emotional capabilities. The contribution of the parent-adolescent relationships to the capacity for closeness and commitment was further mediated through relationships with the best friend, whereas the contribution to the capacity for separateness was not. A substantial number of our participants showed high capability for intimacy although in the military service context the circumstances for the development of intimacy were quite limited and non-optimal. Exploration of the separateness and closeness facets of intimacy in romantic relationships in the two sexes and in other contexts is recommended. PMID- 11476614 TI - The development of commitment and attachment in dating relationships: attachment security as relationship construct. AB - This study examined attachment security and commitment as predictors of relationship stability in a sample of 51 couples. Attachment and commitment were measured during the Fall and Spring semesters prior to graduation from college and relationship stability was determined by contacting couples approximately one year following graduation from college. Attachment security was measured as a relationship construct (appraisal of the dating partner's availability and responsiveness). Attachment security increased with the length of time in the dating relationship and predicted relationship stability following graduation. We then considered how personality and family of origin factors contributed to positive trajectories within the dating relationship (i.e. the growth of attachment security, commitment and relationship stability). Males' security with their mothers and fathers and reports of open communication between their parents contributed to growth in attachment security and commitment between the Fall and Spring semesters. Attachment styles (assessed as a measure of personality) also predicted relationship stability with both males' and females' security increasing the relationship stability one year after graduation. Implications for understanding attachment as both a personality and relationship construct in adult romantic relationships are discussed. PMID- 11476615 TI - Adolescent romance: between experience and relationships. AB - This concluding and integrative paper calls attention to several features and conceptual issues addressed by the contributors of this special issue. The first issue pertains to developmental perspectives in the study of how adolescent romance evolves. The second deals with the various features and concepts of adolescent romance. The third topic discusses the association of adolescent romance with other close relationships occurring during this time span. The fourth topic highlights the importance of the diversity of developmental contexts in shaping romantic relationships. Finally, conceptual issues in the study of adolescent romance are reviewed and the need for future studies of early adolescent romantic experiences is discussed. PMID- 11476616 TI - The resident strikes back: invader-induced switching of resident attractor. AB - The aim of this paper is two-fold: (a) by way of example, we elucidate the phenomenon of invader-induced switches in a resident attractor; (b) we expose in detail how resonance and phase have a strong impact when semelparous organisms (as, e.g. Pacific salmon) with different life-cycle lengths compete in a self induced periodically fluctuating environment. We analyse a simple model for the competition between annuals and biennials, focusing on the situation that the annual population in isolation converges to a two-cycle. Well-timed biennial mutants sample the periodically varying environment more efficiently than the annual resident. They can invade successfully even when they are inferior to the resident, in the sense that they have lower viability and/or fertility. Successful invasion can lead to resonance-mediated coexistence if the invader is rather inferior to the resident. Remarkably, for mutants that are less inferior to the resident, successful invasion by a mutant strategy will inevitably be followed by the extinction of the former invader and concurrent re-establishment of the resident. The expulsion of the invader is brought about by an invasion induced phase shift or attractor switch. We call this phenomenon "the resident strikes back" and say that the resident strategy is invasible, yet invincible. After the resident has struck back, other mutants can successfully invade again. On a longer time-scale, this might lead to an intermittent occurrence of ultimately inferior strategies. The results show that even in a deterministic setting, successful invasion does not necessarily lead to establishment and that mutual invasibility is not always sufficient for coexistence. PMID- 11476617 TI - Topology of an intracellular transduction chain (phototropism of Phycomyces): 1. Joint review of functional, temporal, and spatial aspects. AB - Two light-induced growth reactions in a unicellular cylindrical sporangiophore of Phycomyces blakesleeanus-vertical growth acceleration under symmetrical irradiation (photomecism) and directional growth under unilateral irradiation (phototropism)-share common input light perception as well as common output growth mechanism but have strongly divergent dynamics and other distinctive features. This divergence culminates in the phototropic paradoxes the main of which states that photomecism shows total adaptation, while phototropism does not adapt. The basis for this contradiction is that the phototropic transduction chain, unlike that of photomecism, faces a spatially non-uniform stimulus and processes a series of spatial patterns (light and absorption profiles, adaptation profile, etc.). The only way to resolve the paradoxes and correlate features of both responses within a single transduction chain is to assume non-local signal transduction, e.g. a cross-talk between different azimuthal locations within the cylindrical cell. On the other hand, to establish the presence of an appropriate cross-talk is equivalent of gaining insight into the topology of the transduction chain. This series of two papers contains a review reconsidering the entire field from this viewpoint (Paper 1) and a mathematical model of pattern transduction which unifies features of phototropism and resolves the paradoxes (Paper 2). At the same time, this is the first "proof of concept" for the "activity/pooling (a/p) networks"-a specific mathematical apparatus designed to analyse systemic properties and control in metabolic pathways. PMID- 11476618 TI - Topology of an intracellular transduction chain (phototropism of Phycomyces): 2. Mathematical model of pattern transduction. PMID- 11476619 TI - Probabilities of encounters between objects in biological systems: meta-observer view. AB - Encounters between objects play a crucial role in creating specific patterns of organization in many biological systems. This paper explores a general, quantitative principle of encounter probabilities, applicable over various levels of organization, as a fundamental step toward a comprehensive theory of biological probability ranging from cells through organisms to ecological communities. Based on the cognizers-system model, a general description of encounter probabilities in a finite position space is derived as a function of the number, or density, of objects with cognitive, selectivity properties incorporated as parameters. There is a prevailing wisdom, as supposed in many scientific thoughts, that the per-unit-time number of encounters between focal and target objects is given as a linear function of the density of target objects. One result shows that this idea does not describe the real nature of encounter probabilities or rates, but only a special case or approximation of a fundamental description. The analysis also explicates conditions for ignoring the presence of non-target objects, and those for the aggregation of two or more different objects into the same type sharing the same cognitive properties in the description of the probabilities. PMID- 11476620 TI - Pesticide resistance: can we make it a renewable resource? AB - Negative cross-resistance (NCR) occurs when a mutant allele confers (i) resistance to one toxic chemical and (ii) hyper-susceptibility to another. Sequential deployment of NCR toxins is useful for insect control in few situations (Pittendrigh et al., 2000). Using Monte Carlo simulations, we investigated the concurrent use of a pair of NCR toxins to control a hypothetical insect pest population. When the toxins killed more heterozygotes than homozygotes, the resistance allele became either extremely common or rare depending on starting allelic frequency. If the NCR toxins did not kill the two homozygous groups equally, then the toxin with lesser toxicity eventually played a greater role in the control of the pest population. Based on our results, we present an approach for the systematic development of an NCR toxin after the commercial release of the first toxin. First, large-scale screens are performed to find chemicals that kill the resistant homozygous insects, but not the susceptible ones. Chemicals that preferentially kill resistant insects are then tested for toxicity to the heterozygotes. Those highly toxic to both homo- and heterozygotes are given the highest priority for development. This screen can be adapted to identify compounds useful in controlling antibiotic-, herbicide- or fungicide-resistant organisms. PMID- 11476621 TI - The sense of consciousness. AB - I propose that consciousness might be understood as the property of a system that functions as a sense in the biological meaning of that term. The theory assumes that, as a complex system, the sense of consciousness is not a fixed structure but implies structure with variations and that it evolved, as many new functions do, through the integration of simpler systems. The recognized exteroceptive and enteroceptive senses provide information about the organism's environment and about the organism itself that are important to adaptation. The sense of consciousness provides information about the brain and thus about the organism and its environment. It senses other senses and processes in the brain, selecting and relating components into a form that "makes sense"-where making sense is defined as being useful to the organism in its adaptation to the environment. The theory argues that this highly adaptive organizing function evolved with the growing complexity of the brain and that it might have helped resolve discrepancies created at earlier stages. Neural energies in the brain that are the input to the sense of consciousness, along with the processing subsystem of which they are a part, constitute the base of consciousness. Consciousness itself is an emergent effect of an organizing process achieved through the sense of consciousness. The sense of consciousness thus serves an organizing function although it is not the only means of organization in the brain. Its uniqueness lies in the character of the organization it creates with consciousness as a property of that organization. The paper relates the theory to several general conceptions-interactionism, epiphenomenalism and identity theory-and illustrates a number of testable hypotheses. Viewing consciousness as a property of a sense provides a degree of conceptual integration. Much of what we know about the evolution and role of the conventionally recognized senses should help us understand the evolution and role of the sense of consciousness, and of consciousness itself. PMID- 11476622 TI - Long-term stability of diverse immunological memory. AB - Mathematical modelling is used to examine the effects of T cell apoptosis and memory differentiation dynamics on memory retention (memory stability). Apoptosis by cytokine deprivation induced death, and competition for survival signals are incorporated. Our models indicate that such population dependent processes are essential for the preservation of specific memory to previously encountered pathogens. Memory stability and vaccination properties improve significantly when population dependent mechanisms are present. These mechanisms work synergistically together to further improve memory characteristics. Our models emphasize the role of competitive cellular mechanisms in regulating repertoire structure and characteristics. PMID- 11476623 TI - Cessation of cell proliferation by adjustment of cell redox potential. AB - A variety of chemical substances are known to stop cell proliferation, although the mechanisms are obscure. We suggest that many of these chemicals employ the braking power of the retinoblastoma (RB) protein to stop proliferation by raising the intracellular redox potential. The elevation of the redox potential above a threshold prevents the phosphorylation of RB protein, which in turn impedes the release of transcription factors necessary for the progression of the cell cycle. The redox potential of normal proliferating fibroblasts has been found to be below that of fibrosarcoma cells. The possibility thus exists that, for cells with this property, the dosage of a redox-raising agent can be adjusted to stop cancer-cell proliferation without affecting the proliferation of normal cells. PMID- 11476624 TI - Modelling antipredator vigilance and flight response in group foragers when warning signals are ambiguous. AB - The trade-off between feeding and vigilance in flocks of birds has been extensively studied and modelled. An assumption of many models is that if one bird spots the predator, it gives a signal and the rest of the flock takes flight. However, it has been observed that birds do not always respond to signals and in fact many signals turn out to be false alarms. Since taking flight is both costly in time and energy, it may be advantageous for birds not to respond to all alarm calls. A model is developed to show under what circumstances birds should respond to a signal. The model predicts that under most, but not all, circumstances, birds should respond to multiple detections but not to single detections. The model also predicts that if birds respond to all flights, they will have to compensate for the time lost to feeding and the greater energy requirement of spending more time in flight, by being less vigilant, and they have a lower probability of survival than birds which only respond to multiple detections. PMID- 11476625 TI - The role of T cell help for anti-viral CTL responses. AB - Cytotoxic T lymphocyte (CTL or CD8) responses are a major branch of the immune system involved in controlling viral infections. Murine models have shown that the development of effective and sustained CD8 cell responses requires CD4 T cell help. However, the precise mechanism in which CD4 cells provide help for CD8 cell responses is still controversial. In the literature, mainly two mechanisms are discussed. According to the "classical" pathway, CD4 cells secrete cytokines, such as IL-2, which promote the responsiveness of the CD8 cells. According to the "CD4-APC-CD8" pathway, CD4 cells specifically activate antigen presenting cells (APCs), and APCs specifically interact with CD8 cells, thereby delivering help. Here, we derive kinetic models in order to describe and compare both pathways of help. We find that the two pathways might have different roles in different situations. The classical pathway is more efficient at inducing CD8 cell expansion at high virus loads, while the CD4-APC-CD8 pathway is more efficient at inducing CD8 cell proliferation at low virus loads. From this, it follows that the classical pathway might be needed in order to kick-start a CD8 cell response in the acute phase of the infection, while the CD4-APC-CD8 pathway is needed in order to ensure virus clearance when virus load is reduced by the immune system. These findings have implications for the interpretation of experimental data from virus infection in helper-deficient hosts. In particular, the models offer further suggestions for the development of treatment regimes aimed at achieving immunological control of HIV infection which has been shown to crucially depend on the availability of helper cell responses. PMID- 11476626 TI - Evolution and alignment of the hypervariable arm 1 of Aeschynanthus (Gesneriaceae) ITS2 nuclear ribosomal DNA. AB - Comparative ITS2 sequencing in the plant genus Aeschynanthus(Gesneriaceae) reveals an insertion/deletion (indel) hot spot in the ITS2 sequences that is difficult to align. Examination of other Gesneriaceae sequences shows that this is a widespread phenomenon in this plant family. Minimum free-energy secondary structure analyses localize the hot spot to the terminal part of arm 1. Arm 1 is twice as long in Gesneriaceae than in other asterids. In addition, the pattern of indels is consistent with this secondary structure model. The high variability of the extended terminal part of arm 1 in Gesneriaceae and the fact that it can be deleted altogether imply that it is functionally superfluous. In contrast, the base of arm 1 is relatively conserved and may function as an exonuclease recognition site. This study illustrates how comparative secondary structure analyses can be helpful in fine-scale alignment. Alignment based on secondary structure conflicts with our initial manual alignment and, to a lesser extent, with a CLUSTAL X alignment with default parameters. PMID- 11476627 TI - Phylogeny and evolution of the neotropical rodent genus Calomys: inferences from mitochondrial DNA sequence data. AB - Field mice of the genus Calomys are small, mostly granivorous rodents common to several habitats in South America. To date, phylogenies for the genus have been proposed on the basis of morphological, chromosomal, and biochemical data, often with contradictory results due to incomplete species sampling or methodological shortcomings. In this paper, we propose relationships among 10 species of Calomys based on the complete cytochrome b gene sequence. Our analyses show that Calomys is constituted by two major clades, one mostly associated with mountain habitats with subsequent invasions to lowland habitats and another with species restricted to lowland habitats both north and south of the Amazon basin. The evolution of the genus was likely accompanied by a reduction of chromosome diploid numbers that occurred independently in each of the two evolutionary lineages. A "clock" calibrated on the split between Auliscomys and Loxodontomys suggests that the almost nonexistent fossil record for the genus greatly underestimates divergence times among its species. PMID- 11476628 TI - Phylogeny and evolution of African shrews (Mammalia: Soricidae) inferred from 16s rRNA sequences. AB - Current phylogenetic hypotheses on the African Crocidurinae (Soricidae) are based upon morpho-anatomical, karyological, and allozyme studies. The present study attempts to resolve the interrelationships among African Crocidurinae and their relationships to Eurasian Crocidurinae and to the subfamily Soricinae, on the basis of partial mitochondrial 16s rRNA sequences (549 bp). This is the first molecular study to include all but one of the nine currently recognized African shrew genera. In agreement with current views, two major lineages emerge. The first lineage includes Myosorex and Congosorex and supports the existence of a myosoricine taxon. The second lineage includes the six remaining genera. The genus Sylvisorex appears to be polyphyletic, whereas species of the controversial genus Crocidura are monophyletic. The genus Suncus presumably originated in Africa. The monospecific genera Ruwenzorisorex and Scutisorex and the two representatives of Paracrocidura cluster with species of other genera. Grouping patterns of species from different continents suggest that there have been multiple exchanges between Africa and Eurasia. The time estimates of these exchanges, inferred from two independent fossil-based calibrations of a molecular clock, coincide with the time estimates for migration events in other mammalian taxa. PMID- 11476629 TI - Utility and evolution of cytochrome b in insects. AB - Cytochrome b (cyt-b) is widely used in molecular phylogenetic studies of vertebrate, but not invertebrate, taxa. To determine whether this situation is an historical accident or reflects the utility of cyt-b, we compared the abilities of cyt-b, COI, and one nuclear ribosomal gene region (D1 of 28S) to recover intergeneric relationships within the tiger moth tribes Ctenuchini and Euchromiini. Additionally, we compared the rate of sequence and amino acid evolution of cyt-b across insects. Cytochrome b had the same level of sequence variation and A/T bias as COI, but was less useful for recovering intergeneric relationships. The total evidence tree casts doubt on the traditional taxonomy of the group. For the class Insecta, we found that functional conservation of amino acids occurs for the same regions as those found in vertebrates with the exception of Mallophaga (lice). Lice have an accelerated rate of nonsynonymous substitutions. Accelerated rate of cyt-b nucleotide and amino acid evolution in Apidae (bees) may be correlated with increased metabolic rates associated with facultative endothermy (= heterothermy). PMID- 11476630 TI - Molecular phylogenetics of Australo-Papuan possums and gliders (family Petauridae). AB - Phylogenetic relationships within the possums of the family Petauridae, including their affinities with the family Pseudocheiridae, were inferred from DNA sequences obtained for the mitochondrial ND2 gene (1040 bp) combined with previously published partial 12S rDNA sequences. Short, deep internodes characterize some of the divergences obtained. The robustness of these nodes was assessed by several methods such as exclusion of taxa and partitioning of characters. In all analyses a monophyletic Pseudocheiridae was evident, whereas a monophyletic Petauridae was not as well supported. Within the Petauridae, Gymnobelideus was more closely related to Dactylopsila-Dactylonax than to Petaurus. This supports the results obtained from microcomplement fixation of albumin and DNA-DNA hybridization studies but conflicts with morphological data. PMID- 11476631 TI - Phylogenetic relationships of major clades of Catostomidae (Teleostei: Cypriniformes) as inferred from mitochondrial SSU and LSU rDNA sequences. AB - Suckers (Family Catostomidae) are holarctic in distribution and include 76 recent species in 14 genera, with 13 genera and 75 species occurring in North and Central America and Siberia. Although this group constitutes a significant component of many aquatic ecosystems, most historic systematic effort has been either alpha- or limited beta-level studies focusing on the two largest tribes within the family, the Catostomini and the Moxostomatini. A recent phylogenetic study based on morphological, biochemical, and early life history characters has advanced current understanding of relationships among catostomid fishes. To further examine phylogenetic relationships among basal lineages of catostomids, we sequenced the entire mitochondrial (mt) SSU and LSU rRNA genes from genera representing all subfamilies and tribes within Catostomidae. Phylogenetic analysis of gene sequences yielded monophyletic Catostomidae, Ictiobinae, and Catostominae and para- or polyphyletic Cycleptinae, with Myxocyprinus as the basal-most taxon and Cycleptus as either the next most-basal taxon or the taxon basal to the Catostominae. Relationships within the Catostominae were generally consistent with those proposed in the above-noted recent phylogenetic study although Thoburnia and Hypentelium were either a clade sister to or a grade group relative to Moxostoma and Scartomyzon. In all trees, Scartomyzon was paraphyletic and embedded within Moxostoma. Phylogenetic affinities of Erimyzon and Minytrema varied depending on data set and character weighting scheme employed. To better reflect phylogenetic relationships resolved in this extensive analysis, we propose the following changes to the classification of catostomids: formation of the new subfamily Myxocyprininae, containing Myxocyprinus from China; restriction of the Cycleptinae to the two species of Cycleptus from North America; restriction of the tribe Moxostomatini to Moxostoma and Scartomyzon; Erimyzon and Minytrema are incertae sedis within Catostominae; and resurrection of the tribe Thoburniini, containing Thoburnia and expanded to include Hypentelium. PMID- 11476632 TI - From the Old World to the New World: a molecular chronicle of the phylogeny and biogeography of hystricognath rodents. AB - Hystricognath rodents include Old World Phiomorpha and New World Caviomorpha. These two groups have an enigmatic biogeographical history. Using a nuclear marker, the exon 28 of the von Willebrand Factor gene (vWF), we reconstructed the phylogenetic relationships among 23 Hystricognathi species. These taxa encompass the complete familial diversity of the Hystricognathi. Our results indicate a basal trifurcation of hystricognaths leading to Hystricidae, Phiomorpha s.s. (Bathyergidae, Thryonomyidae, and Petromuridae), and Caviomorpha. The monophyly of caviomorphs is robustly supported, confirming a single colonization event of South America by hystricognaths. Caviomorpha are divided into four lineages: Cavioidea, Erethizontoidea, Chinchilloidea, and Octodontoidea. Furthermore, we suggest that (1) Chinchillidae and Dinomyidae are sister clades, (2) Abrocomidae is a true Octodontoidea, and (3) Capromyidae, Echimyidae, and Myocastoridae cluster together. Surprisingly, Erethizontidae does not appear to be the most diverged caviomorph lineage. The molecular results are discussed in the light of previous paleontological and morphological observations. Local molecular clocks are used to estimate divergence dates among hystricognath lineages. An Asian origin is suggested for Caviomorpha, and a colonization route through Australia and Antarctica is indicated as an alternative to the hypothesis of a transatlantic migration of Caviomorpha from Africa to South America. PMID- 11476633 TI - A phylogeny of freshwater eels inferred from mitochondrial genes. AB - The genus Anguilla Shaw of Family Anguillidae consists entirely of freshwater eels, including 15 species and 2 subspecies. Conventionally, variegated markings and the length of the dorsal fin are the major morphological features used for reconstruction of phylogenetic relationships. The evolutionary history of these species remains unclear, especially for the Atlantic eels, whose habitats are far from the Metropolis in the Indo-Pacific region. This study reexamined the phylogenetic relationships of 12 Anguilla species by sequencing of the cytochrome b and 12S rRNA genes. In our analysis, species bearing similar coloration patterns or dorsal fin morphology are not necessarily clustered in the same clade, indicating that these morphological features might be unstable or might have occurred independently in different lineages during evolution. Combining our molecular data and geographical evidence, we speculate that (1) Anguilla first radiated about 20 million years ago, (2) the ancestors of Atlantic eels did not migrate by drifting through the Tethys Seaway at the leptocephali stage but instead trekked across the Central American Isthmus to the Sargasso Sea for spawning at the adult stage, and (3) multiple radiation events had occurred at the Metropolis during Anguilla evolution. PMID- 11476634 TI - New partial sequences of phosphoenolpyruvate carboxylase as molecular phylogenetic markers. AB - To better understand the evolution of the enzyme phosphoenolpyruvate carboxylase (PEPC) and to test its versatility as a molecular character in phylogenetic and taxonomic studies, we have characterized and compared 70 new partial PEPC nucleotide and amino acid sequences (about 1100 bp of the 3' side of the gene) from 50 plant species (24 species of Bryophyta, 1 of Pteridophyta, and 25 of Spermatophyta). Together with previously published data, the new set of sequences allowed us to construct the up to now most complete phylogenetic tree of PEPC, where the PEPC sequences cluster according to both the taxonomic positions of the donor plants and the assumed specific function of the PEPC isoforms. Altogether, the study further strengthens the view that PEPC sequences can provide interesting information for the reconstruction of phylogenetic relations between organisms and metabolic pathways. To avoid confusion in future discussion, we propose a new nomenclature for the denotation of PEPC isoforms. PMID- 11476635 TI - A mitogenomic perspective on the basal teleostean phylogeny: resolving higher level relationships with longer DNA sequences. AB - A recent study demonstrated that mitochondrial genomic (mitogenomic) data comprising nucleotide sequences from the concatenated protein-coding (no 3rd codon positions) plus transfer RNA (stem regions only) genes reproduced the expected phylogeny of teleosts with high statistical support. We reexamined the interrelationships of the five major, basal teleostean lineages (Osteoglossomorpha, Elopomorpha, Clupeomorpha, Ostariophysi, and Protacanthopterygii; given various rankings) using mitogenomic data for which five alternative phylogenetic hypotheses have been previously proposed on the basis of both morphological and molecular analyses. In addition to previously determined complete mitochondrial DNA (mtDNA) sequences from eight basal teleosts and two outgroups, we determined the complete mtDNA sequences (excluding a portion of the control region) for two, purposefully chosen species of Osteoglossomorpha (Osteoglossum bicirrhosum and Pantodon buchholzi), and the data were subjected to maximumparsimony and maximum-likelihood analyses. The resultant tree topologies from the two methods were congruent, although they differed from any of the previously proposed hypotheses. Furthermore, the mitogenomic data confidently rejected all of these hypotheses with high statistical significance. PMID- 11476636 TI - Phylogenetic analyses of two mitochondrial genes and one nuclear intron region illuminate European subterranean termite (Isoptera: Rhinotermitidae) gene flow, taxonomy, and introduction dynamics. AB - Phylogenetic analyses of multiple DNA sequences were conducted to elucidate gene flow, evolutionary patterns, taxonomy, and the dynamics of two accidental introductions: Reticulitermes lucifugus grassei into Devon, United Kingdom and R. flavipes into Europe. Two mitochondrial DNA genes totaling 1495 bp and a 380-bp ribosomal intergenic transcribed spacer were sequenced. Neighbor-joining and parsimony analyses revealed that multiple female lineages of R. lucifugus grassei were introduced into Devon possibly from southwestern France, where the species was indigenous. The taxonomic status of the European R. santonensis as a species separate from the North American R. flavipes has been questioned since it was described in 1924. Phylogenetic analyses revealed a close genetic relationship between R. flavipes from the United States and R. santonensis from France. These analyses, coupled with morphological and chemotaxonomic data, provide strong support for R. santonensis and R. flavipes being the same species. They also suggested that R. santonensis infestations likely resulted from R. flavipes being introduced into Europe. PMID- 11476637 TI - Molecular phylogeny and biogeography of West Indian toads (Anura: Bufonidae). AB - Toads of the Bufo peltocephalus Group (Anura: Bufonidae) occur throughout the Greater Antilles (Cuba to the Virgin Islands), a geographic region of relatively high endemicity. Previous morphological and immunological studies suggested that the West Indian toads are a monophyletic lineage derived from Neotropical Bufo but were unable to clarify relationships within the group. We examined the evolutionary relationships and biogeography of this group of frogs by collecting approximately 2 kb of mitochondrial DNA sequence data from eight West Indian species and selected non-West Indian species from the New World and the Old World. Our analyses support the monophyly of native West Indian toads and a New World origin for the group. Relationships among the West Indian species are less certain, but a Cuban lineage is defined in most analyses. PMID- 11476638 TI - Evidence for an independent radiation of endosymbiotic litostome ciliates within Australian marsupial herbivores. AB - The recent discovery of isotrichid-like ciliates occurring as endosymbionts in macropodid marsupials posed interesting questions in regard to both their phyletic origin (all previous records confined to eutherian mammals) and their morphological evolution (Australian forms possibly representing missing links between previously described genera). The SSU rRNA gene was sequenced for three species (Dasytricha dehorityi, D. dogieli, and Bitricha tasmaniensis) and aligned against representatives of all major ciliate classes. The Australian species did not group with the other isotrichid species but instead formed an independent radiation. Discrepancies between recent global phylogenies of the phylum Ciliophora were examined by manipulation of the aligned sequence data set. Sources of conflict between these studies did not stem from differences in outgroup choice or phylogenetic reconstruction methods. Differences in the application of confidence limits and primary sequence alignment have probably resulted in the reporting of spurious associations which are not supported by more conservative confidence or alignment methodology. At present, the ciliate subphylum Intramacronucleata is an unresolved polytomy which may be due to deficiencies in the SSU rRNA gene sequence dataset or indicate that the ciliates radiated into their extant classes by rapid burst-like evolution. PMID- 11476639 TI - Re: Phylogenetic relationships in Sphingidae (Insecta: Lepidoptera): initial evidence from two nuclear genes. PMID- 11476640 TI - Chromosomal evolution among six mosquito species (Diptera: Culicidae) based on shared restriction fragment length polymorphisms. PMID- 11476641 TI - Ddc and amd sequences resolve phylogenetic relationships of Drosophila. PMID- 11476643 TI - Stalking disrupts lives, leaves emotional scars: perpetrators are often mentally ill. PMID- 11476644 TI - Psychiatrists explore legacy of traumatic stress in early life. PMID- 11476645 TI - From the Food and Drug Administration. PMID- 11476648 TI - Improving standards in international humanitarian response: the Sphere Project and beyond. PMID- 11476649 TI - Atorvastatin for acute coronary syndromes. PMID- 11476650 TI - Atorvastatin for acute coronary syndromes. PMID- 11476652 TI - Hepatitis B vaccination for newborns. PMID- 11476654 TI - Laboratory containment of wild poliovirus. PMID- 11476655 TI - Imagery rehearsal therapy for chronic nightmares in sexual assault survivors with posttraumatic stress disorder: a randomized controlled trial. AB - CONTEXT: Chronic nightmares occur frequently in patients with posttraumatic stress disorder (PTSD) but are not usually a primary target of treatment. OBJECTIVE: To determine if treating chronic nightmares with imagery rehearsal therapy (IRT) reduces the frequency of disturbing dreams, improves sleep quality, and decreases PTSD symptom severity. DESIGN, SETTING, AND PARTICIPANTS: Randomized controlled trial conducted from 1995 to 1999 among 168 women in New Mexico; 95% had moderate-to-severe PTSD, 97% had experienced rape or other sexual assault, 77% reported life-threatening sexual assault, and 58% reported repeated exposure to sexual abuse in childhood or adolescence. INTERVENTION: Participants were randomized to receive treatment (n = 88) or to the wait-list control group (n = 80). The treatment group received IRT in 3 sessions; controls received no additional intervention, but continued any ongoing treatment. MAIN OUTCOME MEASURES: Scores on the Nightmare Frequency Questionnaire (NFQ), Pittsburgh Sleep Quality Index (PSQI), PTSD Symptom Scale (PSS), and Clinician-Administered PTSD Scale (CAPS) at 3- and 6-month follow-up. RESULTS: A total of 114 participants completed follow-up at 3 and/or 6 months. Comparing baseline to follow-up (n = 97 114), treatment significantly reduced nights per week with nightmares (Cohen d = 1.24; P<.001) and number of nightmares per week (Cohen d = 0.85; P<.001) on the NFQ and improved sleep (on the PSQI, Cohen d = 0.67; P<.001) and PTSD symptoms (on the PSS, Cohen d = 1.00; P<.001 and on the CAPS, Cohen d = 1.53; P<.001). Control participants showed small, nonsignificant improvements for the same measures (mean Cohen d = 0.21). In a 3-point analysis (n = 66-77), improvements occurred in the treatment group at 3-month follow-up (treatment vs control group, Cohen d = 1.15 vs 0.07 for nights per week with nightmares; 0.95 vs -0.06 for nightmares per week; 0.77 vs 0.31 on the PSQI, and 1.06 vs 0.31 on the PSS) and were sustained without further intervention or contact between 3 and 6 months. An intent-to-treat analysis (n = 168) confirmed significant differences between treatment and control groups for nightmares, sleep, and PTSD (all P<.02) with moderate effect sizes for treatment (mean Cohen d = 0.60) and small effect sizes for controls (mean Cohen d = 0.14). Posttraumatic stress symptoms decreased by at least 1 level of clinical severity in 65% of the treatment group compared with symptoms worsening or not changing in 69% of controls (chi(2)(1) = 12.80; P<.001). CONCLUSIONS: Imagery rehearsal therapy is a brief, well-tolerated treatment that appears to decrease chronic nightmares, improve sleep quality, and decrease PTSD symptom severity. PMID- 11476656 TI - Longitudinal study of psychiatric symptoms, disability, mortality, and emigration among Bosnian refugees. AB - CONTEXT: Evidence is emerging that psychiatric disorders are common in populations affected by mass violence. Previously, we found associations among depression, posttraumatic stress disorder (PTSD), and disability in a Bosnian refugee cohort. OBJECTIVE: To investigate whether previously observed associations continue over time and are associated with mortality emigration to another region. DESIGN, SETTING, AND PARTICIPANTS: Three-year follow-up study conducted in 1999 among 534 adult Bosnian refugees originally living in a refugee camp in Croatia. At follow-up, 376 (70.4%) remained living in the region, 39 (7.3%) were deceased, 114 (21.3%) had emigrated, and 5 (1%) were lost to follow up. Those still living in the region and the families of the deceased were reinterviewed (77.7% of the original participants). MAIN OUTCOME MEASURES: Depression and PTSD diagnoses, based on Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria and measured by the Hopkins Symptom Checklist-25 and the Harvard Trauma Questionnaire, respectively; disability, measured by the Medical Outcomes Study Short-Form 20; and cause of death, determined by family interviews with review of death certificates, if available. RESULTS: In 1999, 45% of the original respondents who met the DSM-IV criteria for depression, PTSD, or both continued to have these disorders and 16% of respondents who were asymptomatic in 1996 developed 1 or both disorders. Forty six percent of those who initially met disability criteria remained disabled. Log linear analysis revealed that disability and psychiatric disorder were related at both times. Male sex, isolation from family, and older age were associated with increased mortality after adjusting for demographic characteristics, trauma history, and health status (for male sex, adjusted odds ratio [OR], 2.63; 95% confidence interval [CI], 1.17-5.92; living alone, OR, 2.40; 95% CI, 1.07-5.38; and each 10-year increase in age, OR, 1.91; 95% CI, 1.34-2.71). Depression was associated with higher mortality in unadjusted analysis but was not after statistical adjustment (unadjusted OR, 3.12; 95% CI, 1.55-6.26; adjusted OR, 1.85; 95% CI, 0.82-4.16). Posttraumatic stress disorder was not associated with mortality or emigration. Spending less than 12 months in the refugee camp (OR, 11.30; 95% CI, 6.55-19.50), experiencing 6 or more trauma events (OR, 3.34; 95% CI, 1.89-5.91), having higher education (OR, 1.90; 95% CI, 1.10-3.29), and not having an observed handicap (OR, 0.11; 95% CI, 0.02-0.52) were associated with higher likelihood of emigration. Depression was not associated with emigration status. CONCLUSIONS: Former Bosnian refugees who remained living in the region continued to exhibit psychiatric disorder and disability 3 years after initial assessment. Social isolation, male sex, and older age were associated with mortality. Healthier, better educated refugees were more likely to emigrate. Further research is necessary to understand the associations among depression, emigration status, and mortality over time. PMID- 11476657 TI - Lifetime events and posttraumatic stress disorder in 4 postconflict settings. AB - CONTEXT: Little is known about the impact of trauma in postconflict, low-income countries where people have survived multiple traumatic experiences. OBJECTIVE: To establish the prevalence rates of and risk factors for posttraumatic stress disorder (PTSD) in 4 postconflict, low-income countries. DESIGN, SETTING, AND PARTICIPANTS: Epidemiological survey conducted between 1997 and 1999 among survivors of war or mass violence (aged >/=16 years) who were randomly selected from community populations in Algeria (n = 653), Cambodia (n = 610), Ethiopia (n = 1200), and Gaza (n = 585). MAIN OUTCOME MEASURE: Prevalence rates of PTSD, assessed using the PTSD module of the Composite International Diagnostic Interview version 2.1 and evaluated in relation to traumatic events, assessed using an adapted version of the Life Events and Social History Questionnaire. RESULTS: The prevalence rate of assessed PTSD was 37.4% in Algeria, 28.4% in Cambodia, 15.8% in Ethiopia, and 17.8% in Gaza. Conflict-related trauma after age 12 years was the only risk factor for PTSD that was present in all 4 samples. Torture was a risk factor in all samples except Cambodia. Psychiatric history and current illness were risk factors in Cambodia (adjusted odds ratio [OR], 3.6; 95% confidence interval [CI], 2.3-5.4 and adjusted OR,1.6; 95% CI, 1.0-2.7, respectively) and Ethiopia (adjusted OR, 3.9; 95% CI, 2.0-7.4 and adjusted OR, 1.8; 95% CI, 1.1-2.7, respectively). Poor quality of camp was associated with PTSD in Algeria (adjusted OR, 1.8; 95% CI, 1.3-2.5) and in Gaza (adjusted OR, 1.7; 95% CI, 1.1-2.8). Daily hassles were associated with PTSD in Algeria (adjusted OR, 1.6; 95% CI, 1.1-2.4). Youth domestic stress, death or separation in the family, and alcohol abuse in parents were associated with PTSD in Cambodia (adjusted OR, 1.7; 95% CI, 1.1-2.6; adjusted OR, 1.7; 95% CI, 1.0-2.8; and adjusted OR, 2.2; 95% CI, 1.1-4.4, respectively). CONCLUSIONS: Using the same assessment methods, a wide range of rates of symptoms of PTSD were found among 4 low-income populations who have experienced war, conflict, or mass violence. We identified specific patterns of risk factors per country. Our findings indicate the importance of contextual differences in the study of traumatic stress and human rights violations. PMID- 11476658 TI - Malnutrition, measles, mortality, and the humanitarian response during a famine in Ehiopia. AB - CONTEXT: The World Food Programme estimated that 10 million people were at risk of starvation in Ethiopia in 2000 but later reported that a famine had been averted. However, no population-based data on mortality or nutrition existed for Gode district, at the epicenter of the famine in the Somali region of Ethiopia. OBJECTIVES: To estimate mortality rates, determine the major causes of death, and estimate the prevalence of malnutrition among children and adults for the population of Gode district. DESIGN AND SETTING: Two-stage cluster survey conducted from July 27 through August 1, 2000, which included anthropometric measures and 8-month retrospective mortality data collection. PARTICIPANTS: A total of 595 households comprising 4032 people living in Gode district of Ethiopia. MAIN OUTCOME MEASURES: Crude mortality rates and mortality rates for children younger than 5 years, causes of death, weight for height of less than -2 z scores among children aged 6 months to 5 years, and body mass index of less than 18.5 kg/m(2) among adults and older persons. RESULTS: Of the 595 households, 346 (58.2%) were displaced from their usual places of residence. From December 1999 through July 2000, a total of 293 deaths occurred in the sample population; 159 (54.3%) deaths were among children younger than 5 years and 72 (24.6%) were among children aged 5 to 14 years. The crude mortality rate was 3.2/10 000 per day (95% confidence interval [CI], 2.4-3.8/10 000 per day), which is 3 times the cutoff used to define an emergency. The mortality rate for children younger than 5 years was 6.8/10 000 per day (95% CI, 5.4-8.2/10 000 per day). Approximately 77% of deaths occurred before major relief interventions began in April/May 2000. Wasting contributed to 72.3% of all deaths among children younger than 5 years. Measles alone or in combination with wasting accounted for 35 (22.0%) of 159 deaths among children younger than 5 years and for 12 (16.7%) of 72 deaths among children aged 5 to 14 years. The prevalence rate for wasting (weight for height of <-2 z score) among children aged 6 months to 5 years was 29.1% (95% CI, 24.7% 33.4%). Using a method to adjust body mass index for body shape, the prevalence of undernutrition (body mass index <18.5 kg/m(2)) among adults aged 18 to 59 years was 22.7% (95% CI, 17.9%-27.5%). CONCLUSIONS: To prevent unnecessary deaths, the humanitarian response to famine needs to be rapid, well coordinated, and based on sound epidemiological evidence. Public health interventions, such as mass measles vaccination campaigns with coverage extended to children aged 12 to 15 years should be implemented as the first priority. The prevalence of wasting and undernutrition among children and adults, respectively, should be assessed in all prolonged, severe famines. PMID- 11476659 TI - Dating violence against adolescent girls and associated substance use, unhealthy weight control, sexual risk behavior, pregnancy, and suicidality. AB - CONTEXT: Intimate partner violence against women is a major public health concern. Research among adults has shown that younger age is a consistent risk factor for experiencing and perpetrating intimate partner violence. However, no representative epidemiologic studies of lifetime prevalence of dating violence among adolescents have been conducted. OBJECTIVE: To assess lifetime prevalence of physical and sexual violence from dating partners among adolescent girls and associations of these forms of violence with specific health risks. DESIGN, SETTING, AND PARTICIPANTS: Female 9th through 12th-grade students who participated in the 1997 and 1999 Massachusetts Youth Risk Behavior Surveys (n = 1977 and 2186, respectively). MAIN OUTCOME MEASURES: Lifetime prevalence rates of physical and sexual dating violence and whether such violence is independently associated with substance use, unhealthy weight control, sexual risk behavior, pregnancy, and suicidality. RESULTS: Approximately 1 in 5 female students (20.2% in 1997 and 18.0% in 1999) reported being physically and/or sexually abused by a dating partner. After controlling for the effects of potentially confounding demographics and risk behaviors, data from both surveys indicate that physical and sexual dating violence against adolescent girls is associated with increased risk of substance use (eg, cocaine use for 1997, odds ratio [OR], 4.7; 95% confidence interval [CI], 2.3-9.6; for 1999, OR, 3.4; 95% CI, 1.7-6.7), unhealthy weight control behaviors (eg, use of laxatives and/or vomiting [for 1997, OR, 3.2; 95% CI, 1.8-5.5; for 1999, OR, 3.7; 95% CI, 2.2-6.5]), sexual risk behaviors (eg, first intercourse before age 15 years [for 1997, OR, 8.2; 95% CI, 5.1-13.4; for 1999, OR, 2.4; 95% CI, 1.4-4.2]), pregnancy (for 1997, OR, 6.3; 95% CI, 3.4 11.7; for 1999, OR, 3.9; 95% CI, 1.9-7.8), and suicidality (eg, attempted suicide [for 1997, OR, 7.6; 95% CI, 4.7-12.3; for 1999, OR, 8.6; 95% CI, 5.2-14.4]). CONCLUSION: Dating violence is extremely prevalent among this population, and adolescent girls who report a history of experiencing dating violence are more likely to exhibit other serious health risk behaviors. PMID- 11476660 TI - Mandatory reporting of domestic violence injuries to the police: what do emergency department patients think? AB - CONTEXT: Laws requiring mandatory reporting of domestic violence to police exist in 4 states. Controversy exists about the risks and benefits of such laws. OBJECTIVE: To examine attitudes of female emergency department patients toward mandatory reporting of domestic violence injuries to police and how these attitudes may differ by abuse status. DESIGN, SETTING, AND PARTICIPANTS: Cross sectional survey conducted in 1996 of 1218 women patients (72.8% response rate) in 12 emergency departments in California (a state with a mandatory reporting law) and Pennsylvania (without such a law). MAIN OUTCOME MEASURES: Opposition to mandatory reporting to police and the characteristics associated with this belief. RESULTS: Twelve percent of respondents (n = 140) reported physical or sexual abuse within the past year by a current or former partner. Of abused women, 55.7% supported mandatory reporting and 44.3% opposed mandatory reporting (7.9% preferred that physicians never report abuse to police and 36.4% preferred physicians report only with patient consent). Among nonabused women, 70.7% (n = 728) supported mandatory reporting and 29.3% opposed mandatory reporting. Patients currently seeing/living with partners (odds ratio [OR], 1.5; 95% confidence interval [CI], 1.1-2.0), non-English speakers (OR, 2.1; 95% CI, 1.4 3.0), and those who had experienced physical or sexual abuse within the last year (OR, 2.2; 95% CI, 1.6-2.9) had higher odds of opposing mandatory reporting of domestic violence injuries. There were no differences in attitudes by location (California vs Pennsylvania). CONCLUSIONS: The efficacy of mandatory reporting of domestic violence to police should be further assessed, and policymakers should consider options that include consent of patients before wider implementation. PMID- 11476661 TI - Recognition and treatment of posttraumatic stress disorder. PMID- 11476662 TI - Public health in times of war and famine: what can be done? What should be done? PMID- 11476663 TI - Violence in E-rated video games. AB - CONTEXT: Children's exposure to violence, alcohol, tobacco and other substances, and sexual messages in the media are a source of public health concern; however, content in video games commonly played by children has not been quantified. OBJECTIVES: To quantify and characterize the depiction of violence, alcohol, tobacco and other substances, and sex in video games rated E (for "Everyone"), analogous to the G rating of films, which suggests suitability for all audiences. DESIGN: We created a database of all existing E-rated video games available for rent or sale in the United States by April 1, 2001, to identify the distribution of games by genre and to characterize the distribution of content descriptors associated with these games. We played and assessed the content of a convenience sample of 55 E-rated video games released for major home video game consoles between 1985 and 2000. MAIN OUTCOME MEASURES: Game genre; duration of violence; number of fatalities; types of weapons used; whether injuring characters or destroying objects is rewarded or is required to advance in the game; depiction of alcohol, tobacco and other substances; and sexual content. RESULTS: Based on analysis of the 672 current E-rated video games played on home consoles, 77% were in sports, racing, or action genres and 57% did not receive any content descriptors. We found that 35 of the 55 games we played (64%) involved intentional violence for an average of 30.7% of game play (range, 1.5%-91.2%), and we noted significant differences in the amount of violence among game genres. Injuring characters was rewarded or required for advancement in 33 games (60%). The presence of any content descriptor for violence (n = 23 games) was significantly correlated with the presence of intentional violence in the game (at a 5% significance level based on a 2-sided Wilcoxon rank-sum test, t(53) = 2.59). Notably, 14 of 32 games (44%) that did not receive a content descriptor for violence contained acts of violence. Action and shooting games led to the largest numbers of deaths from violent acts, and we found a significant correlation between the proportion of violent game play and the number of deaths per minute of play. We noted potentially objectionable sexual content in 2 games and the presence of alcohol in 1 game. CONCLUSIONS: Content analysis suggests a significant amount of violence in some E-rated video games. The content descriptors provide some information to parents and should be used along with the rating, but the game's genre also appears to play a role in the amount of violent play. Physicians and parents should understand that popular E-rated video games may be a source of exposure to violence and other unexpected content for children and that games may reward the players for violent actions. PMID- 11476664 TI - Tuberculosis mortality during a civil war in Guinea-Bissau. AB - CONTEXT: Tuberculosis (TB) is an increasing global problem, despite effective drug therapies. Access to TB therapy during conflict situations has not been studied. OBJECTIVE: To determine the effect of irregular TB treatment due to an armed conflict in Guinea-Bissau, West Africa. DESIGN, SETTING, AND PATIENTS: Ongoing retrospective cohort study conducted in the capital city of Bissau among 101 patients with TB who received irregular or no treatment during the civil war (war cohort; June 7-December 6, 1998) and 108 patients with TB who received treatment 12 months earlier (peace cohort; June 7-December 6, 1997) and comparison of an additional 42 patients who had completed treatment before June 6, 1998, and 69 patients who had completed treatment before June 6, 1997. MAIN OUTCOME MEASURE: Mortality rates, compared by irregular (war cohort) vs regular (peace cohort) access to treatment, by intensive vs continuation phase of treatment, and by those who had previously completed treatment for TB. RESULTS: Irregular treatment was associated with an increased mortality rate among patients with TB. The mortality rate ratio (MR) was 3.12 (95% confidence interval [CI], 1.20-8.12) in the war cohort, adjusting for age, sex, human immunodeficiency virus (HIV) infection, residence, and length of treatment. Each additional week of treatment before the war started increased probability of survival by 5% (95% CI, 0%-10%). In the intensive phase of treatment, the adjusted MR was 3.30 (95% CI, 1.04-10.50) and in the continuation phase it was 2.26 (95% CI, 0.33-15.34). Increased mortality among the war cohort was most marked in HIV-positive patients, who had an adjusted MR of 8.19 (95% CI, 1.62 41.25). Mortality was not increased in HIV-positive or HIV-negative patients who had completed TB treatment when the war started. CONCLUSIONS: Interruption of treatment had a profound impact on mortality among patients with TB during the war in Guinea-Bissau. Regular treatment for TB was associated with significantly improved survival for HIV-infected individuals. In emergencies, it is crucial to ensure availability of TB drugs. PMID- 11476669 TI - Development of porcine embryos reconstituted with somatic cells and enucleated metaphase I and II oocytes matured in a protein-free medium. AB - BACKGROUND: Many cloned animals have been created by transfer of differentiated cells at G0/G1 or M phase of the cell cycle into enucleated M II oocytes having high maturation/meiosis/mitosis-promoting factor activity. Because maturation/meiosis/mitosis-promoting factor activity during oocyte maturation is maximal at both M I and M II, M I oocytes may reprogram differentiated cell nuclei as well. The present study was conducted to examine the developmental ability in vitro of porcine embryos reconstructed by transferring somatic cells (ear fibroblasts) into enucleated M I or M II oocytes. RESULTS: Analysis of the cell cycle stages revealed that 91.2 +/- 0.2% of confluent cells were at the G0/G1 phase and 54.1 +/- 4.4% of nocodazole-treated cells were at the G2/M phase, respectively. At 6 h after activation, nuclear swelling was observed in 50.0 88.9% and 34.4-39.5% of embryos reconstituted with confluent cells and nocodazole treated cells regardless of the recipient oocytes, respectively. The incidence of both a swollen nucleus and polar body was low (6.3-10.5%) for all nocodazole treated donor cell regardless of the recipient oocyte. When embryos reconstituted with confluent cells and M I oocytes were cultured, 2 (1.5%) blastocysts were obtained and this was significantly (P < 0.05) lower than that (7.6%) of embryos produced by transferring confluent cells into M II oocytes. No reconstructed embryos developed to the blastocyst stage when nocodazole-treated cells were used as donors. CONCLUSIONS: Porcine M I oocytes have a potential to develop into blastocysts after nuclear transfer of somatic cells. PMID- 11476670 TI - A missense mutation (Q279R) in the fumarylacetoacetate hydrolase gene, responsible for hereditary tyrosinemia, acts as a splicing mutation. AB - BACKGROUND: Tyrosinemia type I, the most severe disease of the tyrosine catabolic pathway is caused by a deficiency in fumarylacetoacetate hydrolase (FAH). A patient showing few of the symptoms associated with the disease, was found to be a compound heterozygote for a splice mutation, IVS6-1g->t, and a putative missense mutation, Q279R. Analysis of FAH expression in liver sections obtained after resection for hepatocellular carcinoma revealed a mosaic pattern of expression. No FAH was found in tumor regions while a healthy region contained enzyme-expressing nodules. RESULTS: Analysis of DNA from a FAH expressing region showed that the expression of the protein was due to correction of the Q279R mutation. RT-PCR was used to assess if Q279R RNA was produced in the liver cells and in fibroblasts from the patient. Normal mRNA was found in the liver region where the mutation had reverted while splicing intermediates were found in non expressing regions suggesting that the Q279R mutation acted as a splicing mutation in vivo. Sequence of transcripts showed skipping of exon 8 alone or together with exon 9. Using minigenes in transfection assays, the Q279R mutation was shown to induce skipping of exon 9 when placed in a constitutive splicing environment. CONCLUSION: These data suggest that the putative missense mutation Q279R in the FAH gene acts as a splicing mutation in vivo. Moreover FAH expression can be partially restored in certain liver cells as a result of a reversion of the Q279R mutation and expansion of the corrected cells. PMID- 11476671 TI - Method for non-invasively recording electrocardiograms in conscious mice. AB - BACKGROUND: The rapid increase in the development of mouse models is resulting in a growing demand for non-invasive physiological monitoring of large quantities of mice. Accordingly, we developed a new system for recording electrocardiograms (ECGs) in conscious mice without anesthesia or implants, and created Internet accessible software for analyzing murine ECG signals. The system includes paw sized conductive electrodes embedded in a platform configured to record ECGs when 3 single electrodes contact 3 paws. RESULTS: With this technique we demonstrated significantly reduced heart rate variability in neonates compared to adult mice. We also demonstrated that female mice exhibit significant ECG differences in comparison to age-matched males, both at baseline and in response to beta adrenergic stimulation. CONCLUSIONS: The technology we developed enables non invasive screening of large numbers of mice for ECG changes resulting from genetic, pharmacological, or pathophysiological alterations. Data we obtained non invasively are not only consistent with what have been reported using invasive and expensive methods, but also demonstrate new findings regarding gender dependent and age-dependent variations in ECGs in mice. PMID- 11476672 TI - Macular translocation with chorioscleral outfolding: an experimental study. AB - PURPOSE: Macular translocation by chorioscleral infolding has been proposed as a surgical intervention for exudative age-related macular degeneration, but the surgery is unpredictable and can be associated with severe complications. We tested a new surgical technique, macular translocation with chorioscleral outfolding secured by neurosurgical clips. METHODS: This was a prospective interventional study in two parts; the first in human cadaver eyes and the second in pigs. Chorioscleral infolding was performed on six human donor eyes, and chorioscleral outfolding was performed on an additional six. The inner surface of the eye wall was measured, and then the fold was unfolded and the distance was measured again. In the second half of the study, macular translocation surgery was performed on 33 pig eyes with one of three sclera shortening methods: 1) a circumferential chorioscleral infolding using 5-0 nylon sutures, 2) a circumferential chorioscleral outfolding using scleral clips, or 3) a radial chorioscleral outfolding using scleral clips. Foveal translocation was measured. RESULTS: The inner wall of the human cadaver eye was shortened in the chorioscleral infolding group by a mean of 1.6 mm, and in the chorioscleral outfolding group by 3.0 mm. In the pig eyes, the fovea was translocated a mean 2377 microm by circumferential suturing, 2582 microm by circumferential clipping, and 3386 microm by radial clipping. Irregular deformation of the globe was more apparent in the circumferential suture group. Undesirable retinal folds often formed after circumferential infolding but not after radial clipping. CONCLUSION: Radial chorioscleral outfolding with clips is more predictable and effective than infolding. It produces more translocation and prevents folds across the fovea, one of the most undesirable complications in macular translocation surgery. PMID- 11476673 TI - Macular translocation with chorioscleral outfolding: a pilot clinical study. AB - PURPOSE: A new surgical technique to translocate the macula was used to treat patients with subfoveal choroidal neovascularization secondary to age-related macular degeneration (AMD). DESIGN: Prospective, interventional case series. METHODS: Twenty-five eyes of 25 patients underwent macular translocation with either circumferential or radial chorioscleral outfolding using three clip sizes: 2-mm, 3-mm, and 4-mm. Postoperative photocoagulation was performed on only those eyes that had an extrafoveal choroidal neovascular membrane following surgery. RESULTS: The surgery successfully displaced the fovea in 22 (88%) of the eyes. The median postoperative foveal displacement was 1142 microm (range 0 to 3200 microm). Patients who had radial outfolding with 4-mm clips had the greatest displacement of the fovea (range 1644 to 3200 microm median 1977 microm). The fovea was successfully displaced to a location outside the choroidal neovascular membrane in 17 (68%) of the 25 eyes. The best-corrected visual acuity improved in 11 eyes (median, 17 letters), remained unchanged in 4 eyes, and decreased in 10 eyes (median, 12 letters). Visual acuity increased by a median of 2 letters. The final best-corrected visual acuity was 20/64 in 3 eyes; 20/80 in 3 eyes; 20/100 in 4 eyes; 20/126 in 4 eyes; 20/200 in 4 eyes; 20/250 in 4 eyes; and 20/400 in 3 eyes. CONCLUSIONS: Macular translocation with radial chorioscleral outfolding using 4-mm clips resulted in the best foveal displacement and improvement in visual function, and was associated with the least amount of vision loss and complications. Further refinements are needed to make this surgical procedure more predictable, and more research (randomized clinical trials) is needed to determine the role of macular translocation in the treatment of subfoveal choroidal neovascularization in patients with AMD. PMID- 11476674 TI - Foveal translocation with scleral imbrication in patients with myopic neovascular maculopathy. AB - PURPOSE: To report our surgical results of foveal translocation with scleral imbrication in patients with myopic neovascular maculopathy. DESIGN: Noncomparative, interventional, consecutive case series. METHODS: Ten eyes of 10 myopic patients with subfoveal neovascular membranes that had undergone foveal translocation with scleral imbrication were recruited for this retrospective study. Inclusion criteria were myopia 6.0 diopters or greater in refractive error (or axial length 26.5 mm or longer), subfoveal choroidal neovascularization, and preoperative best-corrected visual acuity of 20/100 or worse. None of these eyes had undergone prior laser photocoagulation or submacular surgery. The main outcome measures were surgical complications and postoperative visual function. RESULTS: Postoperatively, visual acuity had improved more than 3 lines in the logarithm of minimum angle of resolution (logMAR) measurement in all eyes. The mean preoperative, postoperative best, and final visual acuity were 0.12, 0.59, and 0.51, respectively. Of the 10 eyes, six achieved a postoperative final visual acuity of 20/40 or better. The mean postoperative foveal displacement was 0.78 disk diameter (range, 0.3--1.3 disk diameter). Two patients underwent a reoperation because of insufficient foveal displacement. Furthermore, one of these two patients required a third operation to reduce an excessive retinal fold involving the fovea induced by the second surgery. Of the 10 patients, two noted transient diplopia. This complaint, however, resolved over time as suppression developed. Although unintentional iatrogenic retinal tears formed intraoperatively in two eyes, these were successfully treated without serious complications. Postoperatively, mild retinal pigment epithelial changes were observed in all cases, but none led to significant deterioration of visual acuity during the follow-up period. All patients but one were followed for a minimum of 6 months. CONCLUSIONS: In eyes with myopic neovascular maculopathy, foveal translocation with scleral imbrication may be useful in improving visual acuity. Further refinements in surgical technique and assessment of the long-term complications will be needed to make this procedure safer and more useful. PMID- 11476675 TI - Ultrasound biomicroscopy of conventional and sutureless pars plana sclerotomies: a comparative and longitudinal study. AB - PURPOSE: To report the rate of ultrasonically visible vitreous incarceration and longitudinal changes of incarcerated vitreous in pars plana sclerotomies after conventional suturing or sutureless technique using ultrasound biomicroscopy. METHODS: Twenty-five consecutive eyes (25 patients) undergoing primary three-port pars plana vitrectomy participated. The first 16 pars plana vitrectomies were performed with standard conventional sutured sclerotomies, and the following nine pars plana vitrectomies were performed with modified sutureless sclerotomies. Patient demographics, diagnoses, procedures, and complications were recorded. Each patient had ultrasound biomicroscopy performed 1 week before surgery, and also after surgery at 1 week, 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, and 6 months. Visible vitreous incarceration was graded as 0 to 3. RESULTS: Vitreous incarceration was seen in 41 of 48 sclerotomies (85.4%) in the conventionally sutured group, and in 23 of 27 sclerotomies (85.2%) in the sutureless group, with no significant difference in severity among sclerotomies within each group and between the two groups. There was a significant difference in the rate of vitreous incarceration between diabetic patients with proliferative retinopathy and others (P =.002). No progressive change of visible vitreous incarceration was noted in any eye during the 6-month postoperative period. No sclerotomy-related complications occurred during the study period. CONCLUSIONS: Ultrasound biomicroscopy showed no difference in the amount of visible vitreous incarceration in conventionally sutured or sutureless sclerotomies. There was no visible longitudinal change in the incarcerated vitreous during the 6 months of follow-up in uncomplicated cases. PMID- 11476676 TI - Indocyanine green staining and removal of internal limiting membrane in macular hole surgery: histology and outcome. AB - PURPOSE: To report the surgical technique, outcome, and histologic findings involving indocyanine green staining and removal of internal limiting membrane in primary macular hole surgery. METHODS: Prospectively, consecutive patients with idiopathic macular hole or myopic macular hole with retinal detachment were recruited. After pars plana vitrectomy and epiretinal membrane removal, the internal limiting membrane was stained and removed. The specimens were stained using hematoxylin and eosin and periodic acid-Schiff. Immunohistochemical staining was also performed for glial fibrillary acidic protein, vimentin, type I and type IV collagen, and actin. RESULTS: Among 10 patients (10 eyes) in the study, nine eyes had stage 3 or 4 macular hole. Four of them had chronic macular hole. The tenth patient had retinal detachment resulting from a myopic macular hole. Postoperatively, all cases had closure of macular hole without an elevated edge and the retina was attached. Seven patients had improvement of 2 or more Snellen lines, whereas visual acuity remained the same for the other three patients. In six eyes in which complete histologic examinations were feasible, internal limiting membrane was confirmed and two eyes also had a small amount of epiretinal membrane. Myofibrocytes in internal limiting membrane, either scattered or as a single layer, were found in three cases. CONCLUSIONS: Removal of indocyanine green--stained internal limiting membrane around idiopathic macular hole or myopic macular hole with retinal detachment is confirmed with histology and may contribute to macular hole closure and retinal reattachment. PMID- 11476677 TI - Optical coherence tomography in optic disk pit maculopathy treated by the macular buckling procedure. AB - PURPOSE: To report optical coherence tomography in optic disk pit maculopathy treated with macular scleral buckling procedure. METHODS: Twenty six eyes (26 patients) with optic disk pit maculopathy were treated with macular buckling procedure and evaluated with optical coherence tomography. In group 1, five eyes were evaluated with optical coherence tomography before and after macular buckling procedure. In group 2, 21 eyes were evaluated with optical coherence tomography after macular buckling procedure. With optical coherence tomography, cross-sectional, horizontal retinal images were obtained through the fovea and optic disk pit. RESULTS: In group 1, four of five eyes showed absorption of intraretinal fluid 7 to 9 months after macular buckling procedure. After absorption of fluid, retinal thickness in the foveal center ranged from 128 to 178 microm. In one eye the channel from optic disk to fovea reopened and fluid reappeared. In group 2, 20 of 21 eyes showed absorption of intraretinal schisis and subretinal fluid after macular buckling procedure. After absorption of fluid, retinal thickness in the foveal center ranged from 119 to 216 microm. CONCLUSIONS: In optic disk pit maculopathy, optical coherence tomography enabled morphologic assessment of the retina before and after macular buckling procedure. In the successfully treated cases the permanent closure of the connection between the optic disk pit and the intraretinal schisis was demonstrated. Eventual disappearance of schisis and subretinal fluid was depicted. It was also proved that the thickness of the macula returned to normal and visual acuity improved. PMID- 11476678 TI - Paraoxonase gene polymorphisms and plasma oxidized low-density lipoprotein level as possible risk factors for exudative age-related macular degeneration. AB - PURPOSE: Paraoxonase (E.C.3.1.1.2) is a polymorphic protein shown to prevent low density lipoprotein oxidation. Our purpose is to evaluate the hypothesis that paraoxonase gene polymorphisms and plasma oxidized low-density lipoprotein level play a role in the occurrence of exudative age-related macular degeneration. METHODS: We analyzed paraoxonase genotypes (A/B, Gln-Arg192 and L/M, Leu-Met54) and plasma oxidized low-density lipoprotein levels in 72 unrelated Japanese patients with exudative age-related macular degeneration and compared the results with those of 140 age-matched and sex-matched control subjects. RESULTS: The distribution of paraoxonase 192 and paraoxonase 54 polymorphisms was significantly different between the patients with age-related macular degeneration and control subjects (chi-square = 6.226, P =.0445, and chi-square = 6.863, P =.0323, respectively). The high frequency of the BB genotype at position 192 was observed in the exudative age-related macular degeneration group compared with control subjects (52.8% vs 35.0%, respectively; P =.0127). The high frequency of the LL genotype at position 54 was observed in the patients more than the controls (91.7% vs 77.1%, respectively; P =.0090). The mean (+/- SE) oxidized low-density lipoprotein levels in the patients was significantly higher than in the controls (19.1 +/- 1.0 vs 16.2 +/- 0.6 U/ml, P <.01). CONCLUSION: These results indicate that the paraoxonase gene polymorphisms may represent a possible genetic risk factor for age-related macular degeneration and that increased plasma oxidized low-density lipoprotein may be involved in the pathogenesis of age-related macular degeneration. PMID- 11476679 TI - Laser in situ keratomileusis for treatment of residual myopia after photorefractive keratectomy. AB - PURPOSE: To evaluate the results of laser in situ keratomileusis after photorefractive keratectomy. METHODS: Eighty eyes of 80 patients with residual myopia after photorefractive keratectomy were reoperated with laser in situ keratomileusis. The study was retrospective. Laser in situ keratomileusis was performed using the automated corneal shaper microkeratome and Chiron Technolas 217-C d1 excimer laser. Data measured after laser in situ keratomileusis included uncorrected visual acuity, best-corrected visual acuity, refraction, haze, pachymetry, and keratometry. The follow-up was at least 12 +/- 1.6 months (range, 12 to 15 months). RESULTS: After laser in situ keratomileusis the mean spherical equivalent was -0.24 diopters +/- 0.78. (range, -3 to +1.5) at 12 months, and the mean uncorrected visual acuity was 0.76 diopters +/- 0.24 (range, 0.1 to 1). Sixty-five eyes (81.3%) had various degrees of haze after laser in situ keratomileusis. One eye (1.2%) lost 2 lines of best-corrected visual acuity. CONCLUSIONS: Laser in situ keratomileusis enhancement may be a good alternative to correct residual myopia and astigmatism after primary photorefractive keratectomy. Corneal haze is a common problem in these eyes, and the treatment after laser in situ keratomileusis enhancement should be the same as the treatment after primary photorefractive keratectomy. PMID- 11476680 TI - Objective assessment of transient corneal haze and its relation to visual performance after photorefractive keratectomy. AB - PURPOSE: Photorefractive keratectomy has the potential to cause transient corneal haze. The purpose of this study was to evaluate the relationship between transient corneal haze as measured by an objective means and high and low contrast visual performance. METHODS: In a prospective study, 44 eyes of 28 patients were examined preoperatively and at 1, 3, 6, and 12 months after photorefractive keratectomy. Five laser in situ keratomileusis and two intrastromal corneal ring segments (Intacs [KeraVision, Fremont, CA]) were included for comparison, because these procedures are not expected to cause haze. Haze was measured using a prototype objective hazemeter, TSPC-3, a modification of the Nidek EAS-1000. Visual performance was measured using high-contrast visual acuity and the Rabin Small Letter Contrast Test. RESULTS: Corneal haze was greatest at the 1-month examination and was consistent with a decrease in visual performance on both tests. Corneal haze resolved in 82% of eyes by 10 +/- 4 months after photorefractive keratectomy. However, visual performance had not returned to preoperative levels in 65% and 81% of these eyes on the high-contrast visual acuity test and the Small Letter Contrast Test, respectively. Eyes that underwent laser in situ keratomileusis and Intacs did not develop corneal haze; however, visual decrements were measured. CONCLUSIONS: As a clinical tool, the TSPC-3 hazemeter objectively measures very subtle changes in haze levels. Corneal haze appears to account for only approximately 50% of visual performance changes in the early healing period after photorefractive keratectomy. Other factors, namely topographic abnormalities, are more likely to be an important cause of persistent visual disturbances. PMID- 11476681 TI - Two patterns of opacity in corneal dystrophy caused by the homozygous BIG-H3 R124H mutation. AB - PURPOSE: To investigate the opacity pattern in corneas with an Arg124His (R124H) homozygous mutation of the BIG-H3 gene. METHODS: Slit-lamp examination was performed on eight patients with corneal dystrophy resulting from a genetically confirmed BIG-H3 R124H homozygous mutation. The birthplace of each patient also was determined. RESULTS: Slit-lamp examination disclosed two types of opacity patterns in corneas with the BIG-H3 R124H homozygous mutation. Type I (n = 4) is a spot-like opacity present in the anterior stroma in which the lesions are confluent. Type I is the same pattern that previous reports have shown to be caused by the BIG-H3 R124H homozygous mutation. The type II corneal opacity pattern (n = 4) is a reticular opacity in the anterior stroma with round translucent spaces. Type II opacity has not been reported previously in association with any corneal dystrophy. The patients with the type I opacity do not share a common birthplace; however, interestingly, the patients with the type II opacity traced their origin to Tottori prefecture in western Japan. CONCLUSION: The BIG-H3 homozygous R124H mutation induces the development of two distinct patterns of corneal opacity, the recognition of which can establish an accurate diagnosis of corneal dystrophy caused by the homozygous BIG-H3 R124H mutation independent of genetic analysis. In addition, genetic factors or circumstantial influences other than the gene responsible for the corneal dystrophy may influence the pattern of corneal opacity. PMID- 11476682 TI - Effect of cataract extraction on blue-on-yellow visual field. AB - PURPOSE: It is assumed that cataract can influence the results of blue-on-yellow perimetry. However, actual sensitivity change in blue-on-yellow perimetry before and after cataract surgery has not been fully demonstrated. METHODS: Prospective. SETTING: Institutional. STUDY POPULATION: Twenty-two eyes of 22 consecutive patients without ocular pathology, other than cataract, known to influence visual field undergoing cataract surgery. OBSERVATION PROCEDURES: Both white-on-white (W W) and blue-on-yellow (B-Y) perimetries. MAIN OUTCOME MEASURES: Global indexes including mean deviation and other pertinent data of white-on-white and blue-on yellow perimetries were compared before and after cataract surgery. RESULTS: Mean deviation was improved after cataract surgery in both white-on-white perimetry (from -6.88 dB to -3.36 dB, P <.0001) and blue-on-yellow perimetry (from -12.22 dB to -3.64 dB, P <.0001, paired t test). However, the mean difference between preoperative and postoperative mean deviation in blue-on-yellow perimetry (8.58 +/- 3.96 dB) was significantly higher than that of white-on-white perimetry (3.52 +/- 2.69 dB; P <.0001, unpaired t test). That is, the change in mean deviation was greater in blue-on-yellow perimetry than in white-on-white perimetry by a factor of 2.4 times. CONCLUSION: Cataract causes predominantly a general reduction of sensitivity in both blue-on-yellow and white-on-white perimetries, and the general reduction of blue-on-yellow sensitivity is far greater than that of white-on-white sensitivity. PMID- 11476683 TI - Results of viscocanalostomy for primary open-angle glaucoma. AB - PURPOSE: Viscocanalostomy is a nonperforating filtering surgical procedure that may avoid postoperative complications common with standard trabeculectomy. This study was conducted to determine the surgical outcome of this procedure after a postoperative observation period of 12 to 36 months. METHODS: Interventional consecutive case series. In a prospective study, a consecutive series of 67 eyes of 67 patients with chronic primary open-angle glaucoma underwent VCS. Excluded were patients with angle closure glaucoma, post-traumatic, uveitic, neovascular, or dysgenetic glaucoma, as well as patients who needed combined cataract-glaucoma procedures. The patients were examined postoperatively on the first day, first week, at 1 month, and then at 3-month intervals through 36 months. At each visit, best-corrected visual acuity, intraocular pressure (IOP), and the appearance of the surgical wound, anterior chamber, and indirect funduscopy were recorded. RESULTS: Complete success was defined as IOP less than or equal to 20 mm Hg and greater than or equal to 30% IOP reduction without medical or additional surgical treatment compared with the IOP from a preoperative level with maximum tolerated medical therapy; qualified success IOP less than or equal to 20 mm Hg with treatment or an IOP reduction less than 30% from preoperative level with maximum tolerated medical therapy; and qualified failure of an IOP greater than 20 mm Hg with glaucoma medication, but no optic nerve or visual field deterioration and complete failure as an eye requiring further glaucoma surgery or lost visual function. The overall success rate was 88% at 1 year, 90% at 2 years, and 88% at 3 years, with a complete success of 68% at 1 year, 60% at 2 and 59% at 3 years. Four eyes had a perforation of the Descemet membrane, three of those needed peripheral iridectomy; six eyes had a microperforation not needing a peripheral iridectomy. Five eyes presented hyphema. Eight eyes presented deterioration of visual function after surgery: three eyes because of corneal astigmatism, one eye because of cataract, and four presented glaucomatous deterioration of the visual field despite pressures under 20 mm Hg. Four eyes had progressive cataract formation judged as independent from surgery. CONCLUSION: VCS provides an overall success rate of 88% and a complete success of 59% 3 years after surgery. The major immediate complication is perforation of the Descemet membrane with a need for peripheral iridectomy. No serious long-term complications were noted in our small series. Visual function remained stable in 55 eyes (82%). PMID- 11476684 TI - Mini-trabeculectomy as initial surgery for medically uncontrolled glaucoma. AB - PURPOSE: To report on the surgical outcome after at least 12 months of follow-up of mini-trabeculectomy (without radial incisions) as an initial surgery. METHODS: In a prospective, institutional study, mini-trabeculectomy was performed as the initial surgical therapy on 41 eyes with medically uncontrolled glaucoma of 41 consecutive patients aged 40 years or older. Mini-trabeculectomy consists of a 3 mm fornix-based conjunctival flap, sclerostomy at 1 mm from the limbus, and a sclerocorneal tunnel without radial incisions. Of the 41 eyes, four eyes did not complete 12 months of follow-up. Of the remaining 37 eyes, one eye underwent cataract extraction 5 months postoperatively and was evaluated only for surgical complications. Thirty-six patients with a mean age of 70.3 +/- 7.4 standard deviation years (range, 54 to 87) completed 12 months or more of follow-up. RESULTS: In the 36 eyes, mean preoperative intraocular pressure was 30.2 +/- 9.3 mm Hg (range, 19 to 54) with 3.0 +/- 1.2 hypotensive medications (range, 1 to 4). After postoperative mean follow-up of 25.0 +/- 9.2 months (range, 12 to 43), intraocular pressure was at or below the planned target intraocular pressure in 35 eyes (97.2%; P <.0001) and mean intraocular pressure was 16.0 +/- 2.8 mm Hg (range, 11 to 21) with 0.8 +/- 1.0 medications (range, 0 to 3). Postoperative complications in the 37 eyes included early postoperative aqueous leakage with moderately shallow or deep anterior chamber in two eyes (5.4%) and rapid cataract progression in one eye (2.7%). The four eyes that were excluded from the study had intraocular pressures of 10 to 16 mm Hg with 0 to 1 hypotensive medications, 3 to 9 months after surgery, respectively. CONCLUSIONS: Mini-trabeculectomy, which may offer clinical and technical advantages over the standard trabeculectomy, was generally efficacious and relatively safe, based on outcome observed at a mean of 25 months follow-up. A controlled study is required to confirm these observations. PMID- 11476685 TI - Comparison of topical brinzolamide 1% and dorzolamide 2% eye drops given twice daily in addition to timolol 0.5% in patients with primary open-angle glaucoma or ocular hypertension. AB - PURPOSE: The aim was to compare topical brinzolamide 1% twice daily with dorzolamide 2% twice daily, each given with timolol 0.5% twice daily, for safety and effects on intraocular pressure in patients with primary open-angle glaucoma or ocular hypertension. METHODS: This double-blind, randomized, active controlled, parallel group study was conducted multinationally at 31 sites, in 241 patients as above, with assessments at baseline and monthly during 3 months of treatment. The primary end point was a diurnal reduction of trough/peak intraocular pressure from a timolol 0.5% twice daily baseline. RESULTS: Both treatment regimens reduced intraocular pressure significantly at all time points (P <.001): brinzolamide plus timolol by -3.6 to -5.3 mm Hg (-14.2 to -21.9%), dorzolamide plus timolol by -3.6 mm Hg to -5.1 mm Hg (-14.1 to -21.2%). Clinically relevant intraocular pressure reductions (decreases 5 mm Hg or greater or absolute intraocular pressure values 21 mm Hg or less) were manifested by 50.0% to 89.3% of patients under brinzolamide plus timolol and by 43.9% to 85.4% under dorzolamide plus timolol. The treatments were equivalent in mean intraocular pressure-lowering. In general, both regimens were well tolerated. However, more patients (P =.001) experienced at least one adverse event with dorzolamide plus timolol (32.8%) as compared with brinzolamide plus timolol (14.7%); also, more patients (P =.001) experienced ocular discomfort (stinging and burning) after dorzolamide plus timolol (13.1%) than after brinzolamide plus timolol (1.7%). CONCLUSIONS: In terms of intraocular pressure reduction, brinzolamide 1% twice daily was equivalent to dorzolamide 2% twice daily, each added to timolol 0.5% twice daily, but brinzolamide produced significantly less ocular burning and stinging. PMID- 11476686 TI - Endogenous mycotic endophthalmitis: variations in clinical and histopathologic changes in candidiasis compared with aspergillosis. AB - PURPOSE: To describe clinical and/or histopathologic features that could help distinguish endogenous Candida endophthalmitis from endogenous Aspergillus intraocular inflammation and to provide histologic documentation of intraocular spread of these agents. METHODS: Twenty-five patients who underwent enucleation, 13 with morphologic features and/or positive culture for Aspergillus and 12 with histologic evidence and/or positive culture for Candida were included in the study. Clinical information was sought from each case. Patients with AIDS were excluded. The enucleated globes were analyzed to detect location of the fungi, vascular invasion by these agents, and inflammatory response. RESULTS: Candida endophthalmitis was noted in patients with a history of gastrointestinal surgery, hyperalimentation, or diabetes mellitus, whereas aspergillosis was present in patients who had undergone organ transplantation or cardiac surgery. Histopathologically, the vitreous was the primary focus of infection for Candida, whereas subretinal/subretinal pigment epithelium infection was noted in eyes with aspergillosis. Retinal and choroidal vessel wall invasion by fungal elements was noted in cases of aspergillosis but not in cases with candidiasis. Both infectious agents induced suppurative nongranulomatous inflammation. CONCLUSIONS: Unlike Candida endophthalmitis, aspergillosis clinically presents with extensive areas of deep retinitis/choroiditis, and vitreous biopsy may not yield positive results. Histopathologically, it appears that Aspergillus grows preferentially along subretinal pigment epithelium and subretinal space. This intraocular infection is usually associated with a high rate of mortality caused by cerebral and cardiac complications. PMID- 11476687 TI - Management of the corneal flap in laser in situ keratomileusis after previous radial keratotomy. AB - PURPOSE: To report a method of managing the corneal flap in patients having laser in situ keratomileusis for the treatment of residual refractive errors after radial keratotomy. DESIGN: Retrospective case series. METHODS: Intraoperative dehiscence of radial keratotomy wounds occurred in 7 eyes of 6 patients treated with laser in situ keratomileusis for residual myopia or astigmatism or hyperopia 5 to 15 years after radial keratotomy. To minimize extension of these tears, the flap was initially rolled like a carpet toward the hinge before the ablation and then rolled away from the hinge to its original position after the ablation. RESULTS: Using this method of managing the laser in situ keratomileusis flap in patients with previous radial keratotomy, all eyes had successful laser in situ keratomileusis, with 1-year postoperative uncorrected visual acuity ranging between 20/16 and 20/25. No eye had loss of spectacle-corrected vision or interface epithelial ingrowth. CONCLUSIONS: Laser in situ keratomileusis has proved to be an effective treatment for correction of residual refractive errors after radial keratotomy. The surgical technique used in these cases was targeted at minimizing shearing forces in lifting the corneal flap to avoid extension of radial keratotomy wound dehiscence, which could lead to epithelial ingrowth and loss of best-corrected vision. PMID- 11476688 TI - Intraocular lens power calculations using corneal topography after photorefractive keratectomy. AB - PURPOSE: To report two patients (two eyes) with previous photorefractive keratectomy, who subsequently underwent cataract extraction years later. DESIGN: Case reports. METHODS: Corneal topography was used to determine corneal power used in intraocular lens power calculations. RESULTS: In two eyes of two patients, intraocular lens calculations after photorefractive keratectomy were inadequate, which resulted in a hyperopic postoperative refractive error requiring implantation of a piggyback intraocular lens. CONCLUSION: Corneal topography to determine corneal power in patients with previous photorefractive keratectomy may result in unpredictable intraocular lens power calculations. The clinical history method is the standard to determine corneal power and should be considered in intraocular lens calculations before cataract surgery. We recommend supplying refractive patients with preoperative data for use in future formulas for intraocular lens selection. PMID- 11476689 TI - Different recurrence patterns after phototherapeutic keratectomy in the corneal dystrophy resulting from homozygous and heterozygous R124H BIG-H3 mutation. AB - PURPOSE: To report the recurrence pattern of corneal deposits after phototherapeutic keratectomy in patients with corneal dystrophies resulting from homozygous and heterozygous Arg124His (R124H) mutation of the BIG-H3 gene. METHODS: Slit-lamp examination was performed on patients with corneal dystrophy resulting from a genetically confirmed BIG-H3 R124H mutation. RESULTS: The recurrence of corneal deposits after phototherapeutic keratectomy in patient with heterozygous R124H mutation was mild; the granular opacities occurred as spot lesions in the central cornea. The patient with a homozygous mutation had a more severe pattern, and the recurrent lesions were diffuse and occurred in the periphery between the corneal epithelium and laser-ablated stroma. The recurrence free interval in homozygous patients was shorter. CONCLUSION: The mutation genotype of BIG-H3 gene determines the recurrence pattern after phototherapeutic keratectomy. PMID- 11476690 TI - Serratia Marcescens corneal ulcer as a complication of orthokeratology. AB - PURPOSE: To report a case of Serratia Marcescens corneal ulcer as a complication of orthokeratology treatment. METHODS: Case report. RESULTS: A 9-year-old male who underwent orthokeratology treatment for 6 months suffered from a corneal ulcer. The refractive state of lesion eye was -5.5D/-1.25D x 180 degrees, and visual acuity was hand motion at 30 cm. He wore a retainer lens, rigid gas permeable lens, overnight for 2 months before the corneal ulcer occurred. Ulcer became worse after tobramycin and gentamycin treatment for 2 days. After ciprofloxacin treatment, the ulcer healed and visual acuity recovered to 20/20 with spectacle correction. Cultures of the cornea tissue and contact lens storage solution both grew Serratia Marcescens, which was sensitive to ciprofloxacin. CONCLUSION: Overnight wearing of a rigid contact lens is a risk factor for a corneal ulcer. PMID- 11476691 TI - Electroretinogram in the diagnosis of endophthalmitis after intraocular lens implantation. AB - PURPOSE: To determine whether single flash electroretinograms can assist in the diagnosis of eyes with endophthalmitis after intraocular lens implantation. METHODS: Sixteen eyes with postoperative endophthalmitis treated with vitrectomy or intraocular lens removal were studied. The preoperative electroretinogram findings, duration until the onset of endophthalmitis, culture results, and postoperative visual acuity were analyzed. RESULTS: Four eyes with a b-wave/a wave (b/a) ratio less than 1.0 and early onset of endophthalmitis within 1 week had worse postoperative visual acuity (no light perception to 20/200) than the other 12 eyes (20/100 to 20/15) (P =.0035). Pseudomonas aeruginosa and Enterococcus faecalis were cultured in these 4 eyes. CONCLUSION: The combination of a b/a ratio less than 1.0 and early onset of endophthalmitis may indicate poor prognosis in a patient with postoperative endophthalmitis. PMID- 11476692 TI - Optic disk edema as a presentation of Propionibacterium acnes endophthalmitis. AB - PURPOSE: To report a patient with an initial presentation of optic disk edema secondary to Propionibacterium acnes endophthalmitis. METHODS: Interventional case report. The patient had surgery and treatment related to endophthalmitis. Nine months after cataract surgery with posterior chamber lens implant, a 79-year old man receiving topical corticosteroids in the same eye presented with best corrected visual acuity of 20/60, a superior visual field defect, and optic nerve edema without intraocular inflammation. One year postoperatively, the eye presented a further decrease in best-corrected visual acuity, and a dense white plaque was noted on the posterior capsule. RESULTS: One year postoperatively, a partial capsulectomy with vitrectomy and injection of intravitreal antibiotics was performed. Hematoxylin and eosin stain of the posterior capsule plaque revealed gram-positive coccobacilli characteristic of P. acnes. CONCLUSION: Propionibacterium acnes endophthalmitis may present after cataract surgery with intraocular lens implant with decreased vision, optic disk edema, and a visual field defect. PMID- 11476693 TI - Late onset lens particle glaucoma as a consequence of spontaneous dislocation of an intraocular lens in pseudoexfoliation syndrome. AB - PURPOSE: To report acute onset lens particle glaucoma associated with pseudoexfoliation-related dislocation of an intraocular lens implant 12 years after cataract surgery. METHODS: Case report. RESULTS: An 80-year-old woman presented with acute onset of left eye pain that was associated with white fleck like particles circulating in the anterior chamber and an intraocular pressure of 48 mm Hg. The posterior chamber intraocular lens within the capsular bag was positioned more posteriorly to the iris plane than usual, and pseudoexfoliative material was present on the lens capsule and the zonules. Anterior chamber aspirate confirmed the presence of lens cortical fibers. CONCLUSION: Spontaneous dislocation of the posterior chamber intraocular lens in patients with pseudoexfoliation syndrome several years after cataract surgery may liberate lens cortical material, causing lens particle glaucoma. PMID- 11476694 TI - Macular hole surgery in an eye with an optic pit. AB - PURPOSE: To report an eye with a full-thickness macular hole and an associated optic pit and the noteworthy intraoperative findings. METHODS: Case report. A 56 year-old woman presented with visual acuity LE: 20/100, a full thickness macular hole, and an optic pit. Optical coherence tomography and ophthalmic examination were performed preoperatively and postoperatively. RESULTS: Although usually a macular hole associated with an optic pit tends to be a lamellar and characterized by outer layer defects within preexisting macular detachments or schisis-like cavities, this type of macular hole was not presented in this case. Although the macular hole resembled the idiopathic type on clinical examination as well as on optical coherence tomography, it could only be closed in the third surgical attempt after using silicone oil as a long-standing tamponade. Peeling of an epiretinal membrane or the internal limiting membrane was not possible during any of the three surgeries. CONCLUSION: Our observations suggest that in cases of macular hole in association with optic pit, instillation of silicone oil should be considered in the first surgical procedure, especially if no epiretinal membrane or internal limiting membrane peeling is possible intraoperatively. PMID- 11476695 TI - Optical coherence tomography evaluation of idiopathic macular hole treatment by gas-assisted posterior vitreous detachment. AB - PURPOSE: To report a case of idiopathic macular hole, with vitreoretinal traction confirmed by optical coherence tomography that was successfully treated by a single intravitreous perfluoropropane (C(3)F(8)) gas bubble injection. METHODS: Case report. A 65-year-old patient with idiopathic macular hole (stage 2, one eye) received an intravitreous gas injection and was prospectively followed with optical coherence tomography. RESULTS: A complete posterior vitreous detachment was achieved within 6 weeks after gas injection. Visual acuity improved from 20/80 to 20/25 by 10 months of follow-up. Optical coherence tomography disclosed vitreoretinal traction release and macular hole closure. No complications were related to the procedure. CONCLUSION: This simple procedure can assist a complete posterior vitreous detachment with relief of the hyaloid-foveolar traction, facilitating macular hole closure. PMID- 11476696 TI - Simultaneous bilateral central retinal vein occlusion associated with anticardiolipin antibodies in leukemia. AB - PURPOSE: To present a dramatic case of simultaneous bilateral central retinal vein occlusion associated with leukemia and anticardiolipin autoantibodies. METHODS: Interventional case report. Clinical examination, fluorescein angiography, B-mode ultrasonography, and laboratory serologies were performed on a 65-year-old man with chronic myelogenous leukemia who presented with sudden onset of decreased vision in his right eye. RESULTS: Fundus examination disclosed a nonischemic central retinal vein occlusion. Over the next 3 weeks, a nonperfused central retinal vein occlusion developed in both eyes and subsequent neovascular glaucoma developed in his left eye. Throughout this time, his white blood cell and platelet counts remained normal and his serum viscosity remained low, but anticardiolipin antibodies were increased. CONCLUSION: This case reveals the occurrence of simultaneous bilateral central retinal vein occlusion associated with anticardiolipin antibodies in leukemia and suggests an additional mechanism other than hyperviscosity for bilateral central retinal vein occlusions in leukemic patients. PMID- 11476697 TI - Bilateral central retinal vein occlusion in Eisenmenger syndrome. AB - PURPOSE: To report a case of bilateral central retinal vein occlusion in a patient with Eisenmenger syndrome. METHODS: Case report. A 60-year-old woman with Eisenmenger syndrome secondary to a congenital ventricular septal defect presented with a 2-week history of decreased vision in both eyes. RESULTS: Ophthalmic examination revealed bilateral central retinal vein occlusion. Laboratory findings included hematocrit of 50.3% and pO(2) of 52 mm Hg while on O(2) 5 L/min per nasal cannula, despite having undergone phlebotomy 2 weeks earlier. CONCLUSION: Chronic hypoxia caused by Eisenmenger syndrome may result in polycythemia with resultant hyperviscosity and bilateral central retinal vein occlusion. PMID- 11476698 TI - Central retinal artery occlusion in association with an aneurysm of the internal carotid artery. AB - PURPOSE: To report an unusual case of central retinal artery occlusion in a 41 year-old woman. DESIGN: Interventional case report. METHODS: A 41-year-old woman presented with sudden and painless decrease of visual acuity of the left eye. Indirect ophthalmoscopy revealed the typical fundus findings of a central retinal artery occlusion. The fovea was spared by a cilioretinal arteriole. RESULTS: Cerebral angiography showed an aneurysm of the left internal carotid artery, measuring approximately 1.7 x 1.5 x 1.7 cm. The aneurysm was located in close proximity to the ophthalmic branch. CONCLUSION: Aneurysm of the intracranial arteries should be included in the differential diagnosis of central retinal artery occlusion in young patients. PMID- 11476699 TI - The relation of coffee and caffeine to the 5-year incidence of early age-related maculopathy: the Beaver Dam Eye Study. AB - PURPOSE: To examine the relationship between coffee and caffeine consumption and the 5-year incidence of early age-related maculopathy and its component lesions, soft indistinct drusen or pigmentary abnormalities. DESIGN: A prospective cohort study conducted from 1988 to 1995 with an average follow-up of 4.8 years. METHODS: Data from baseline and 5-year follow-up examinations were analyzed for Beaver Dam Eye Study participants (n = 3435). The Wisconsin Age-Related Maculopathy Grading System was used to assess the presence and severity of early age-related maculopathy. RESULTS: Men were more likely to be coffee and caffeine drinkers than were women. For both men and women, coffee and caffeine intake decreased with age. Coffee and caffeine consumption were not associated with the 5-year incidence of early age-related maculopathy, soft indistinct drusen, or pigmentary abnormalities. CONCLUSION: Neither a history of coffee nor caffeine consumption is associated with incident early age-related maculopathy. PMID- 11476700 TI - Retinal findings in melanoma-associated retinopathy. AB - PURPOSE: To report unusual fundus findings in two cases of melanoma-associated retinopathy. METHODS: Observational case reports. The histories of two patients with melanoma-associated retinopathy were reviewed. Sera from both patients were examined for antibodies against retinal bipolar cells. Immunofluorescence was performed on cryostat sections of unfixed normal human retinas. Sera and IgG from both patients were tested against a known melanoma-associated retinopathy patient as well as a control subject. RESULTS: Our patients had metastatic, cutaneous melanoma and a clinical syndrome consistent with melanoma-associated retinopathy. Both patients had serum antibodies that were reactive against retinal bipolar cells. They had unusual fundus changes not previously described in association with melanoma-associated retinopathy. One patient also developed vitiligo. CONCLUSION: Patients with metastatic cutaneous melanoma and melanoma-associated retinopathy may present unusual fundus lesions that may be caused by autoimmunity which is part of the clinical picture of melanoma-associated retinopathy or metastatic melanoma. PMID- 11476701 TI - Masquerade syndrome: T-cell prolymphocytic leukemia presenting as panuveitis. AB - PURPOSE: To report a case of T-cell prolymphocytic leukemia with panuveitis as the primary presenting feature. METHODS: Case report. RESULTS: A 46-year-old woman presented with pain and blurred vision in the right eye. She was found to have signs of panuveitis with a central exudative retinal detachment. Further investigations revealed that she was suffering from the rare T-cell prolymphocytic leukemia. Both systemic and ocular manifestations of the disease resolved after chemotherapy with Campath-IH antigen and as she went into complete remission. The exudative detachment settled, and visual acuity recovered to 20/20. CONCLUSION: This case illustrates that leukemias can present with primarily ocular findings, and the sudden appearance of a serous retinal detachment with inflammatory signs in an otherwise healthy person warrants a thorough systemic screening for an underlying malignancy. PMID- 11476702 TI - Congenital intraocular teratoma associated with eyelid coloboma. AB - PURPOSE: To describe the first case of intraocular teratoma associated with eyelid coloboma and the second reported case of intraocular teratoma. DESIGN: Interventional case report. METHODS: A left intraocular tumor was surgically resected from a 2-day-old female with an associated lower eyelid coloboma. RESULTS: Pathologic evaluation revealed a completely intraocular tumor comprising derivatives of all three germ cell layers giving a diagnosis of intraocular teratoma. The eyelid coloboma was repaired, and a scleral-wrapped hydoxyapatite integrated orbital implant was placed. CONCLUSION: To our knowledge, this is the second reported instance of teratoma originating within the globe and the only reported case of teratoma associated with eyelid coloboma. Although exceedingly rare, intraocular teratoma should be added to the differential diagnosis of congenital intraocular tumors. PMID- 11476703 TI - Management of eyelid anomalies associated with Blepharo-cheilo-dontic syndrome. AB - PURPOSE: To describe the previously unreported management of the eyelid anomalies associated with blepharocheilo-dontic syndrome. Blepharo-cheilo-dontic syndrome is a syndrome of congenital cleft lip/palate, oligodontia, euryblepharon, eyelid ectropion, and lagophthalmos. METHODS: Case report. A 3-month-old male presented with bilateral upper and lower eyelid ectropion with eyelid retraction, marked euryblepharon, severe lagophthalmos, and a right side cleft lip/palate. Lateral tarsal strips, lower eyelid retractor disinsertion, myocutaneous advancement of the cheek and eyelids, and lateral tarsorrhaphy were performed. RESULTS: Correction of the ectropion, eyelid retraction, euryblepharon, and marked improvement in his lagophthalmos were achieved postoperatively. No recurrence of the ectropion or euryblepharon has been noted after 6 months of follow-up. CONCLUSION: Appropriate reconstructive surgery of the eyelids reduces the morbidity associated with the eyelid anomalies and provides an excellent cosmetic result for patients with blepharo-cheilo-dontic syndrome. PMID- 11476704 TI - Fat adherence syndrome after retinal surgery treated with amniotic membrane transplantation. AB - PURPOSE: We present favorable results with amniotic membrane transplantation in a patient who developed fat adherence syndrome after retinal surgery. DESIGN: Interventional case report. METHODS: A 37-year-old man had diplopia resulting from hypotropia of the left eye after retinal detachment surgery. Removal of a previously implanted silicon sponge had little effect because of fibrous adhesion between the inferior rectus muscle and adjacent periorbital fat. We performed amniotic membrane transplantation combined with conventional extraocular muscle surgery. RESULTS: Postoperatively, supraduction of the left eye became almost full. The field of binocular vision was extended markedly by treatment, both in the primary position and with downward gaze. These improvements remained stable over 1 year of follow-up. CONCLUSION: Amniotic membrane transplantation appears to be effective for preventing regrowth of restrictive scar tissue in the fat adherence syndrome. PMID- 11476705 TI - Skew deviation as a complication of cardiac catheterization. AB - PURPOSE: To report three patients who developed diplopia and skew deviation after cardiac catheterization. DESIGN: Interventional case series. METHODS: Three patients complained of diplopia after cardiac catheterization for myocardial infarction (two male patients) or aortic dissection (one female patient). Examination demonstrated skew deviation in each patient. RESULTS: Diplopia and skew deviation were mild and resolved completely in 4 months (case 1), 16 months (case 2), and 1.5 months (case 3). Sensitive signs of minor ischemic damage to the brain stem that cause such disorders are not detectable by neuroimaging. CONCLUSION: Rarely, cardiac catheterization may be complicated by diplopia and skew deviation. PMID- 11476706 TI - Optic tract compression from dolichoectatic basilar artery. AB - PURPOSE: To report a case of optic tract compression caused by a dolichoectatic basilar artery. DESIGN: Observational case report. METHODS: A 74-year-old man with progressive loss of vision over 13 years and no other neurologic signs or symptoms was found to have bilateral optic nerve head pallor and a left homonymous hemianopia. RESULTS: Magnetic resonance imaging and angiography revealed a severe dolichoectatic basilar artery compressing the right optic tract. CONCLUSION: Basilar artery dolichoectasia may rarely cause compression of the optic tract and progressive visual loss. PMID- 11476707 TI - Acute asymmetric upper nasal quandrantanopsia caused by a chiasmal colloid cyst in a patient with multiple sclerosis and bilateral retrobulbar neuritis. AB - PURPOSE: To report a patient with multiple sclerosis and a history of sequential bilateral retrobulbar neuritis, who developed new onset of highly asymmetric upper quadrantanopsia. DESIGN: Interventional case report. METHOD: A 36-year-old woman with multiple sclerosis and bilateral retrobulbar neuritis developed an acute asymmetric upper nasal quadrantanopsia. RESULTS: Magnetic resonance imaging of the brain revealed a cyst that caused chiasmal compression and bilateral visual field defects. CONCLUSION: New onset of bilateral visual field defects in a patient with diagnosed multiple sclerosis is likely to be caused by a new attack of the demyelinating disease. In this case, a newly diagnosed chiasmal colloid cyst was the cause of visual field defects. PMID- 11476728 TI - Antidiuretic action of vasopressin: quantitative aspects and interaction between V1a and V2 receptor-mediated effects. AB - (1) Vasopressin (VP), or antidiuretic hormone, is secreted in response to either increases in plasma osmolality (very sensitive stimulus) or to decreases in plasma volume (less sensitive stimulus). Its normal plasma level is very low (about 1 pg/ml, i.e. 10(-12) M), close to the detection limit of present immunoassays, and distinct antidiuretic effects are observed after infusion of small undetectable amounts of VP. (2) This antidiuretic action results from three main effects of VP on principal cells of the collecting duct (CD) mediated by occupancy of peritubular V2 receptors. (i) Increase in water permeability along the entire CD (via AQP2). (ii) Increase in urea permeability in only the terminal inner medullary CD (via UT-A1). (iii) Stimulation of sodium reabsorption, mainly in the cortical and outer medullary CD (via ENaC). VP also acts on medullary vasculature (V1a receptors) to reduce blood flow to inner medulla without affecting blood flow to outer medulla. Besides these actions, all concurring to increase urine osmolality in different and additive ways, other VP effects, probably exerted through V1a receptors located on luminal membrane, tend to limit the antidiuretic effects of the hormone. They induce the formation of prostaglandins which reduce V2-dependent cAMP accumulation in these cells and thus partially inhibit all three V2 effects. (3) Because urine is first diluted along the nephron before being concentrated in the medulla, VP is required, not only for urine concentration, but first for re-equilibration of tubular fluid osmolality with plasma osmolality, a step taking place in the renal cortex, and achieved through the reabsorption of large quantities of water (more than what is subsequently reabsorbed in the medulla to concentrate urine). Accordingly, VP effects on urine flow-rate are not linear. Small changes in plasma VP in the low range of urine osmolality will induce wide changes in urinary flow-rate, whereas in the upper range of urine osmolality larger changes in plasma VP induce much more limited further reduction in urine flow-rate. (4) Most likely, the different effects of VP require different levels of VP concentration to occur and are thus recruited successively with progressive rise in VP secretion. PMID- 11476729 TI - Vasopressin V2 receptor antagonists. AB - Hyponatremia due to the syndrome of inappropriate antidiuretic hormone secretion (SIADH) and disorders of water retention such as congestive heart failure and cirrhosis is a common problem encountered in the care of the medical patient. Thus far, available treatment modalities for disorders of excess arginine vasopressin (AVP) secretion or action have been suboptimal. The development of nonpeptide AVP V2 receptor antagonists represents a promising treatment option to directly antagonize the effects of elevated plasma AVP concentrations by increasing the water permeability of renal collecting tubules, thereby promoting excretion of retained water and normalizing hypoosmolar hyponatremia. In this review, SIADH and other water retaining disorders are briefly discussed, after which the published preclinical and clinical studies in the development of several nonpeptide AVP V2 receptor antagonists are summarized. The likely therapeutic indications and potential complications of these compounds, as well as their vascular effects, are also described. PMID- 11476730 TI - The role of V2 vasopressin antagonists in hyponatremia. AB - Hyponatremia is a frequent electrolyte disorder. It is often found in congestive cardiac failure, liver cirrhosis, plasma volume contraction and in SiADH. In these disorders hyponatremia is caused by nonosmotic vasopressin and sustained fluid intake. This provides a rationale for V2 vasopressin receptor antagonists in the treatment of hyponatremia. There is now convincing evidence from different animal models of congestive cardiac failure that peptide and non-peptide V2 vasopressin antagonists effectively increase renal water diuresis and plasma sodium concentration. In addition, several of the experimental studies also showed an improvement of hemodynamic changes of cardiac failure in response to V2 antagonists. Data in patients indicated that oral non-peptide V2-antagonists correct hyponatremia and may improve hemodynamic derangements in cardiac failure. In addition, experimental and clinical studies of V2 antagonists have been undertaken in liver cirrhosis and SiADH. In those studies hyponatremia was improved or corrected, too. Taken together, V2 vasopressin antagonists promise to become therapeutic agents in hyponatremic disorders. PMID- 11476731 TI - The V2 vasopressin receptor mutations and fluid homeostasis. AB - Although three different G-protein coupled receptors have been identified for arginine vasopressin, a significant physiological role has been recognized only for the V2 subtype that controls water homeostasis. Identification of the gene encoding the V2 vasopressin (or antidiuretic hormone) receptor enabled researchers to test the hypothesis that mutations of this gene were responsible for X-linked recessive nephrogenic diabetes insipidus. The affected patients are unable to concentrate their urine and as a consequence live in constant danger of dehydration that can cause death, particularly in infancy, or lead to severe hypernatremia that can impair their intellectual and physical development. The danger of severe dehydration diminishes in the adult patients, although they remain highly susceptible to this condition for the rest of their lives. PMID- 11476732 TI - Atrial natriuretic peptide mimetics and vasopeptidase inhibitors. AB - There is now substantial evidence supporting a role of the natriuretic peptides as a major defence mechanism against excess salt and water retention and high blood pressure. Because of this there has been considerable interest in the therapeutic potential of the natriuretic peptide system. Several approaches have been explored including the use of native peptides, the development of natriuretic peptides mimetics and targetting of endogenous clearance of natriuretic peptides. While ANP and BNP administration may be valuable in some circumstances, however, the limitations of the use of peptides especially for long-term treatment are well apparent. In view of this, considerable effort has been devoted to the development of orally active agents to enhance endogenous natriuretic peptides through inhibition of breakdown by neutral endopeptidase. This research has now led to the vasopeptidase inhibitors - dual inhibitors of both endopeptidase and angiotensin converting enzyme. These agents clearly provide a novel approach to enhance endogenous natriuretic peptide function on a background of reduced angiotensin II activity and may lead to an important advance in the treatment of hypertension and of conditions associated with overt salt and water overload. PMID- 11476733 TI - Molecular biology of natriuretic peptides and nitric oxide synthases. AB - Natriuretic peptides and nitric oxide play important roles in cardiovascular and renal physiology and disease. The natriuretic peptides - atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide - comprise a family of proteins that participate in the integrated control of intravascular volume and arterial blood pressure. The natriuretic peptides differentially bind distinct classes of receptors that signal through different mechanisms. Membrane bound, guanylyl cyclase-coupled natriuretic peptide receptors (A- and B-types) mediate natriuretic peptide effects through the production of 3',5'-cyclic guanosine monophosphate (cGMP). C-Type natriuretic peptide receptors, which lack the guanylyl cyclase domain, alter target cell function through G(i) protein coupled inhibition of membrane adenylyl cyclase activity, and also serve to clear circulating natriuretic peptides. The expression of the natriuretic peptides and their receptors are subject to complex controls. Similar structural and regulatory diversity exists for the nitric oxide synthases. The three nitric oxide synthase genes are regulated by a variety of mechanisms ranging from alternative splicing and alternative promoter usage to complex post-translational controls. This review highlights the molecular diversity of the natriuretic peptides and nitric oxide synthases and explores recent insights into their regulation. PMID- 11476734 TI - Plasma A- and B-type natriuretic peptides: physiology, methodology and clinical use. PMID- 11476735 TI - The renal urodilatin system: clinical implications. AB - A renal natriuretic peptide and the 'renal urodilatin system' were identified after the observation that immunoassayable ANP in urine may not be identical to the circulating cardiac hormone ANP, which is a peptide of 28 amino acids. Urodilatin (INN: Ularitide) is a natriuretic peptide isolated from human urine and belongs to the family of A-type natriuretic peptides. Urodilatin is differentially processed to a peptide of 32 amino acids from the same precursor as ANP. It is synthesized in kidney tubular cells and secreted luminally. After secretion from epithelial cells of the distal and/or connecting tubules, Urodilatin interacts downstream at distal segments of the nephron with luminally located receptors whereby it regulates Na(+) and water reabsorption. Thus, the physiological function of the renal Urodilatin system can be described as a paracrine intrarenal regulator for Na(+) and water homeostasis, considering Urodilatin as a real diuretic-natriuretic regulatory peptide. However, the regulation upon which the Urodilatin secretion depends is still not clear. Since Urodilatin has been discovered, a great number of pharmacological and clinical investigations have been carried out using Urodilatin as a drug for several indications. So far, clinical phase I and II studies for acute renal failure, congestive heart failure, and bronchial asthma have been performed. PMID- 11476736 TI - Regulation of mesangial cell function by vasodilatory signaling molecules. AB - Proliferation of mesangial cells and expansion of mesangial matrix is a hallmark of glomerular disease leading to end-stage renal failure and requiring renal replacement therapy. Independently from the type of injury, e.g. in glomerulonephritis or diabetic nephropathy, the response to injury is remarkably uniform. Chronic glomerular disease is frequently associated with increases in systemic blood pressure and altered intraglomerular hemodynamics. Furthermore, reduction of systemic blood pressure and inhibition of the vasoconstrictor peptide angiotensin II have been shown to delay end-stage renal failure in various types of human kidney disease. Since vasoconstrictors of mesangial cells and efferent glomerular arterioli, such as angiotensin II, are thought to be detrimental for the progression of chronic glomerular disease, we propose that vasodilatory factors which antagonize the effects of angiotensin II, might have beneficial effects during the course of progressive kidney disease. To support this concept we will summarize currently available data on the role of vasodilatory signaling molecules such as natriuretic peptides (ANP, BNP and CNP), nitric oxide (NO), the prostaglandines PGE2 and prostacycline, and the purine mediator adenosine in the regulation of mesangial function. PMID- 11476737 TI - Physiological effects of vasopressin and atrial natriuretic peptide in the collecting duct. AB - Vasopressin plays a primary role in the concentration of urine to maintain body fluid homeostasis. The collecting duct as well as thick ascending limb is a major target site of vasopressin. The antidiuretic action of vasopressin is mediated by the V2 receptor in the basolateral membrane of principal cells in the collecting ducts. The binding of vasopressin to V2 receptors causes an activation of adenylate cyclase and a synthesis of cAMP. Vasopressin regulates water and ion transport through V2 receptor-mediated ion channels and transporters. In contrast, the V1a receptor mainly in the luminal membrane of distal nephron regulates basolateral V2 receptor-mediated action with regard to water and ion transport through the activation of G(q/11) and phosphoinositide turnover. Guanylate cyclase forms three types of ANP receptors, although NPR-A and B (GC-A and B) are biologically active and related to the synthesis of cGMP. Urodilatin, synthesized by the kidney, causes natriuresis by binding to GC-A in the collecting ducts. ANP causes diuresis and natriuresis, at least in part by inhibiting the V2 receptor-mediated action of AVP in the collecting ducts. The site of interaction of ANP and AVP is post cAMP synthesis, at least in the collecting ducts. The roles of AVP and ANP under pathophysiological conditions have been reported. PMID- 11476738 TI - Endothelin as a natriuretic hormone: the case for a paracrine action mediated by nitric oxide. PMID- 11476739 TI - The role of the natriuretic peptides in the cardiovascular system. AB - The discovery of the natriuretic peptide family was a breakthrough in modern cardiovascular physiology as it provided a direct link between the heart and the kidneys in the regulation of natriuresis. Along with vasopressin and the renin angiotensin-aldosterone system, the natriuretic peptides comprise the key peptides on which our present understanding of neuroendocrine regulation of the cardiovascular system is based. Three natriuretic peptides have been identified; the A-type, B-type and C-type natriuretic peptides. The former two, the A- and B type natriuretic peptides, function mainly in the cardiovascular system and comprise the cardiac natriuretic peptides. Together with our increased understanding of the neurohormonal regulation of the cardiovascular system in recent years, the discovery of the natriuretic peptide family was important in the establishment of the new field of cardiovascular endocrinology. PMID- 11476740 TI - Water and sodium regulation in chronic heart failure: the role of natriuretic peptides and vasopressin. PMID- 11476741 TI - Therapeutic benefits of increasing natriuretic peptide levels. AB - Natriuretic peptides play an important role in water and salt homeostasis and in the regulation of the cardiovascular system. In recent years, exogenous administration of natriuretic peptides has primarily been used to improve our understanding of the role of natriuretic peptides. Also, it became evident that natriuretic peptides may be used therapeutically. Because of their peptide character, they cannot be administered orally and, therefore, may be used for short-term intravenous therapy only. In recent years, inhibitors of neutral endopeptidase, which degrades natriuretic peptides to inactive metabolites, have been investigated. This review focuses on the potential benefits of increasing natriuretic peptide levels, either through exogenous administration or inhibiting the degradation of endogenous natriuretic peptides. PMID- 11476742 TI - Cardiac natriuretic peptides during exercise and training after heart transplantation. PMID- 11476743 TI - Employing vasopressin during cardiopulmonary resuscitation and vasodilatory shock as a lifesaving vasopressor. AB - Epinephrine during cardiopulmonary resuscitation (CPR) is being discussed controversially due to its beta-receptor mediated adverse effects such as increased myocardial oxygen consumption, ventricular arrhythmias, ventilation perfusion defect, postresuscitation myocardial dysfunction, ventricular arrhythmias and cardiac failure. In the CPR laboratory simulating adult pigs with ventricular fibrillation or postcountershock pulseless electrical activity, vasopressin improved vital organ blood flow, cerebral oxygen delivery, resuscitability, and neurological recovery better than did epinephrine. In paediatric preparations with asphyxia, epinephrine was superior to vasopressin, whereas in both paediatric pigs with ventricular fibrillation, and adult porcine models with asphyxia, combinations of vasopressin and epinephrine proved to be highly effective. This may suggest that a different efficiency of vasopressors in paediatric vs. adult preparations; and different effects of dysrhythmic vs. asphyxial cardiac arrest on vasopressor efficiency may be of significant importance. Whether these theories can be extrapolated to humans is unknown at this point in time. In patients with out-of-hospital ventricular fibrillation, a larger proportion of patients treated with vasopressin survived 24 h compared with patients treated with epinephrine; during in-hospital CPR, comparable short term survival was found in groups treated with either vasopressin or epinephrine. Currently, a large trial of out-of-hospital cardiac arrest patients being treated with vasopressin vs. epinephrine is ongoing in Germany, Austria and Switzerland. The new CPR guidelines of both the American Heart Association, and European Resuscitation Council recommend 40 U vasopressin intravenously, and 1 mg epinephrine intravenously as equally effective for the treatment of adult patients in ventricular fibrillation; however, no recommendation for vasopressin was made to date for adult patients with asystole and pulseless electrical activity, and paediatrics due to lack of clinical data. When adrenergic vasopressors were unable to maintain arterial blood pressure in patients with vasodilatory shock, continuous infusions of vasopressin ( approximately 0.04 to approximately 0.1 U/min) stabilised cardiocirculatory parameters, and even ensured weaning from catecholamines. PMID- 11476744 TI - Potentiation of urinary atrial natriuretic peptide interferes with macula densa function. AB - OBJECTIVES: Neutral endopeptidase (NEP) inhibition potentiated the renal action of Atrial Natriuretic Peptide (ANP) and was associated with appearance of the peptide in the urine, providing evidence of protection of the filtrated peptide along the course of the nephron. The macula densa, composed of epithelial cells, receives ionic information from the urinary compartment via Na-K-2Cl cotransport and influences renin secretion by the myoepithelioid cells in the afferent arteriole. bNOS constitutively expressed in the epithelial cells of the macula densa is involved in this feed-back. NEP inhibition was associated with the absence of any increase in renin secretion. The hypothesis is that potentiation of urinary ANP by NEP inhibition could limit renin secretion by directly or indirectly targeting the macula densa in vivo. METHODS AND RESULTS: We tested the interaction between NEP inhibition (candoxatril) and Na-K-2Cl inhibition (bumetanide) on electrolyte and ANP urinary excretion, renin secretion, macula densa activity (NADPH diaphorase activity and bNOS mRNA) and TSC-1 mRNA expression in the renal cortex and BSC-1 in the renal medulla of rats treated for 5 days. Bumetanide increased urinary electrolyte excretion whereas candoxatril did not. Candoxatril increased urinary ANP and cyclic GMP excretion. Bumetanide increased renin and aldosterone secretion whereas candoxatril decreased renin secretion. This effect on renin release was associated with an increase in macula densa NADPH diaphorase activity in the bumetanide-treated group which was blunted by candoxatril. Lastly, bumetanide increased TSC-1 mRNA expression in the cortex and this effect was blunted by candoxatril. CONCLUSION: These results suggest that potentiation of ANP by NEP inhibition could interfere with tubular function at different levels and limit renin secretion by a urinary pathway involving macula densa activity. PMID- 11476745 TI - Cellular localization, membrane distribution, and possible function of guanylyl cyclases A and 1 in collecting ducts of rat. AB - BACKGROUND: Natriuretic peptides regulate Na+ and H(2)O transport in the cortical collecting duct (CCD). We have shown that natriuretic peptides have no effect on ion conductances or water transport of principal cells (PC) even though a cGMP regulated K+ channel is located in the basolateral membrane of these cells. METHODS: RT-PCR was used to screen for different guanylyl cyclases (GC) in CCD and to look for the expression of GC-1 and GC-A mRNA in CCD of male and female Wistar and Sprague-Dawley rats. Polyclonal antibodies were raised against the detected GC. BCECF was used to investigate the effects of ANP on intracellular pH in intercalated cells (IC). RESULTS: GC-A and GC-1 were detected. GC-A was immunolocalized in the luminal membrane of IC while GC-1 was mainly found in the luminal membrane of PC. GC-1 is expressed in Sprague-Dawley and Wistar rats except for male Sprague-Dawley rats, while GC-A is expressed in all strains. ANP (160 nM, n=11), urodilatin (140 nM, n=6), which had no effect in PC, significantly decreased pH(i) by 0.02+/-0.01 and 0.03 +/- 0.01 Units in IC, respectively. ANP as well as urodilatin and 8-Br-cGMP decreased the pH(i) recovery after acidification by 30 +/- 6% (n=12), 37 +/- 7% (n=8), and 19 +/- 3% (n=8), respectively. CONCLUSION: GC-A is located in the luminal membrane of IC of rat CCD and ANP acts through this receptor when regulating pH(i) via an inhibition of the Na+/H+-exchanger. PC do not possess GC-A. GC-1 seems to be the only GC in these cells of most rat strains tested and therefore, it could be responsible for the regulation of K+ channels in the basolateral membrane via cGMP-dependent protein kinase. PMID- 11476746 TI - Impaired angiotensin II regulation of renal C-type natriuretic peptide mRNA expression in experimental diabetes mellitus. AB - OBJECTIVE: Abnormalities in the regulation of natriuretic peptides (NP) associated with major diseases such as hypertension, heart failure, and diabetes mellitus (DM) have been reported. We investigated levels of mRNA for the vasodilator C-type natriuretic peptide (CNP) in the renal cortex of streptozotocin (STZ)-diabetic rats and the influence of an angiotensin II inhibition. METHODS: DM was induced in Wistar rats by a single STZ injection. Rats were kept for 12 weeks. Additionally, the influence of the ACE inhibitor ramipril (Ram: 3 mg/kg/day) and the AT1 receptor antagonist losartan (Los: 20 mg/kg/day) on CNP expression in the STZ-diabetic and control groups was studied (each group n=6). Animals were characterized by their mean arterial blood pressure, plasma glucose levels, and renal function (each group n=9). After extraction of total renal cortical RNA, CNP expression was analyzed by Northern blots. RESULTS: Renal function was impaired in STZ-diabetic rats which has been improved by Ram and Los treatment. Untreated STZ-diabetic rats showed no difference in renal CNP expression compared to untreated controls. Ram and Los treatments led to an increase in renal cortical CNP mRNA in both diabetic and non diabetic rats. This effect was weaker in STZ-diabetic rats (Ram: control 5.4 fold, STZ 3.5-fold; Los: control 4.2-fold, STZ 1.9-fold). CONCLUSION: These results clearly demonstrate a direct regulatory effect of the renin-angiotensin system (RAS) on renal mRNA levels of CNP. We suggest that RAS inhibition not only prevents the generation of angiotensin II (AngII) but also leads to a stimulation of CNP expression. We conclude that AngII suppresses CNP expression via the AT1 receptor and this mechanism is impaired in STZ-diabetic rats. PMID- 11476747 TI - Temporal changes in natriuretic and antinatriuretic systems after closure of a large arteriovenous fistula. AB - OBJECTIVE: Surgical closure of a large arteriovenous (A-V) fistula in patients and animals is associated with prompt diuresis and natriuresis. However, the mechanisms underlying these changes remained largely unknown. METHODS: The present study evaluated the hormonal balance between major antinatriuretic systems (plasma renin activity, PRA, and arginine vasopressin, AVP) and natriuretic systems (atrial natriuretic peptide, ANP, and renal nitric oxide, NO) in Wistar rats with an A-V fistula (1.2 mm O.D., side to side) between the abdominal aorta and inferior vena cava. RESULTS: The placement of an A-V fistula caused progressive sodium retention (UNaV decreased from 1500 to 100 microequiv./day), a significant drop in mean arterial blood pressure (MAP) from 127+/-3 to 75+/-2 mmHg (P<0.01), and a significant increase in ANP (from 94+/-12 to 389+/-135 pg/ml, P<0.05), PRA (from 22.1+/-2.0 to 47+/-14 ng angiotensin I [Ang I]/ml/h, P<0.05), AVP (from 14.2+/-3.6 to 37.7+/-9.6 pg/ml, P<0.05), norepinephrine (from 184.2+/-40.5 to 1112.6+/-293.2 pg/ml, P<0.05) and epinephrine (from 667.5+/-175.9 to 2049.8+/-496.9 pg/ml, P<0.05). Furthermore, these changes were associated with a 3-fold increase in the renal medullary immunoreactive levels of endothelial NO synthase (eNOS), an endogenous vasodilator that plays an important role in the regulation of medullary blood flow. After 6 days, rats with A-V fistula and maximal sodium retention underwent surgical closure of the A-V fistula. The A-V fistula closure was associated with dramatic natriuresis (UNaV=2563+/-78 and 1918+/-246 microEq/day on days 3 and 6 following the closure, respectively) and restoration of MAP to normal levels (111+/-6 mmHg); PRA decreased to 29+/-5 ng Ang I/ml/h, AVP to 20.3+/-7.1 pg/ml, and medullary eNOS declined to basal levels, whereas plasma ANP concentrations remained elevated (380+/-90 pg/ml) after 3 days and returned to normal (92+/-12 pg/ml) on day 6. CONCLUSIONS: These results demonstrate that the creation of A-V fistula is associated with activation of both natriuretic and antinatriuretic systems. Closure of A-V fistula is characterized by shifting the balance in favor of the natriuretic substances. Moreover, the observed alterations in medullary eNOS following the creation and closure of A-V fistula suggest that this system, an important determinant of medullary blood flow, may contribute significantly to the regulation of sodium excretion in this model. PMID- 11476748 TI - Relaxant effect of C-type natriuretic peptide involves endothelium and nitric oxide-cGMP system in rat coronary microvasculature. AB - OBJECTIVE: Recent evidence suggests a possible role for nitric oxide (NO) in atrial natriuretic peptide-induced blood pressure effects. We tested the hypothesis that C-type natriuretic peptide (CNP)-mediated relaxation of the rat coronary circulation involves NO and activation of soluble guanylyl cyclase. METHODS: Rat hearts (n=6 per group) were perfused in vitro at constant flow and the effect of CNP (0.1-3 micromol/l) on coronary perfusion pressure (a measure of vascular tone) and release of guanosine 3',5'-cyclic monophosphate (cGMP) was determined in absence and presence of the nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine (L-NNA; 0.2 mmol/l) or the natriuretic peptide receptor antagonist HS-142-1 (50 microg/ml). The involvement of Ca(2+)-gated and ATP dependent K(+) channels in CNP-induced relaxation was tested with iberiotoxin (30 nmol/l) and glibenclamide (1 micromol/l), respectively. Rings of rat aorta (n=12) were tested using the organ bath set-up. RESULTS: CNP reduced perfusion pressure from 134 +/- 2 mmHg (baseline) to 71 +/- 1 mmHg (-48%) and this effect was significantly attenuated by L-NNA (-37%) or HS-142-1 (-19%). In presence of glibenclamide, CNP reduced perfusion pressure to 92 +/- 2 mmHg (-32%), in presence of iberiotoxin to 93 +/- 1 mmHg (-30% and in their combined presence to 102+/-2 mmHg (-23%) (P<0.05 vs. corresponding control). Basal release of cGMP was increased up to 4-fold by CNP and this increase was reduced (-50%) in presence of L-NNA or HS-142-1 (-68%). By contrast, relaxation of rat aortic rings mounted in organ baths was insensitive to inhibition by L-NNA. CONCLUSION: Relaxation of the coronary resistance vessels of the rat by CNP is partly mediated by the NO-cGMP pathway. These novel data support the existence of an endogenous link between soluble and particulate guanylyl cyclases in the control of natriuretic peptide mediated coronary resistance vessel function. PMID- 11476749 TI - Measurement of plasma brain natriuretic peptide level as a guide for cardiac overload. AB - OBJECTIVES: We examined whether measurement of the plasma BNP concentrations might be useful for the early diagnosis of the existence and severity of disease in patients with heart disease in daily clinical practice. METHODS AND RESULTS: The plasma BNP and ANP concentrations in 415 patients with heart disease and hypertension and 65 control subjects were measured. Patients with heart disease had higher plasma BNP and ANP concentrations than did those with hypertension or control subjects. Among the etiology of cardiac diseases, specifically dilated cardiomyopathy and hypertrophic cardiomyopathy, was associated with the highest plasma BNP concentrations, whereas dilated cardiomyopathy was associated with the highest plasma ANP concentrations. Plasma BNP concentrations showed an increase as the severity of the heart disease, as graded according to the NYHA classification of cardiac function, increased. In both patients with heart disease and hypertension, the plasma BNP values were higher in those who had abnormalities in their echocardiogram and electrocardiogram as compared to those without any abnormalities. The plasma BNP levels also showed a significant correlation with left ventricular wall thickness and left ventricular mass. On the other hand, the plasma ANP levels showed significant correlations with left ventricular dimension. Receiver operative characteristic analysis revealed that plasma BNP levels showed substantially high sensitivity and specificity to detect the existence of heart diseases. CONCLUSION: Measurements of the plasma BNP concentrations is useful to detect the existence of the diseases, and abnormalities of left ventricular function and hypertrophy in patients with heart disease in daily clinical practice. PMID- 11476750 TI - Intravenous administration of the natriuretic peptide urodilatin at low doses during coronary reperfusion limits infarct size in anesthetized pigs. AB - OBJECTIVE: It has been shown that cGMP content is reduced in post-ischemic myocardium, and that stimulation of cGMP synthesis prevents cardiomyocyte hypercontracture and cell death in vitro. This study was aimed at determining whether administration of the natriuretic peptide urodilatin (URO) at the time of reperfusion could limit myocardial cell death secondary to transient coronary occlusion. METHODS: The relation between cGMP content in reperfused myocardium and the extent of cell death was investigated in isolated rat hearts (n=62) receiving different URO concentrations during initial reperfusion. The dose of intravenous URO necessary to obtain the targeted increase in cGMP in reperfused myocardium was investigated in ten pigs submitted to transient coronary occlusion (CO), and the effect of two selected doses of URO on infarct size was investigated in 22 pigs. RESULTS: cGMP was severely reduced in post-ischemic rat hearts. Addition of 0.01 microM URO during the first 15 min of reperfusion had no effect on myocardial cGMP content, functional recovery or LDH release in hearts submitted to 40 or 60 min of ischemia. At 0.05 microM, URO increased myocardial cGMP to 111% of values in normoxic hearts, improved functional recovery (P=0.01) and reduced peak LDH released by 40% (P=0.02). The beneficial effect of urodilatin was abolished by ANP receptor inhibition. At 1 microM, URO increased cGMP in reperfused myocardium to 363% of normoxic controls and had no beneficial effect. In pigs allocated to 47 min of CO and 5 min of reperfusion, cGMP was markedly reduced in reperfused myocardium. Intravenous URO at 10 ng/kg per min during the first 25 min of reperfusion normalized myocardial cGMP after 5 min of reflow (95% of control myocardium), and reduced infarct size by 40% (P=0.04). At 50 ng/kg per min, urodilatin increased myocardial cGMP in reperfused myocardium to 335% of control myocardium and failed to significantly reduce infarct size (46 vs. 66%, P=0.125). None of these doses had detectable hemodynamic effects. CONCLUSIONS: Intravenous low-dose URO at the time of reperfusion normalizes myocardial cGMP and limits necrosis. Large doses of URO increasing myocardial cGMP well over normal values may lack this beneficial effect. PMID- 11476751 TI - Differential expression of cardiac ANP and BNP in a rabbit model of progressive left ventricular dysfunction. AB - OBJECTIVE: Activation of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) is considered a hallmark of myocardial remodeling. To determine magnitude and relative proportion of activation during the progression to heart failure, we assessed ANP and BNP gene expression in atrial and left ventricular (LV) tissue in a newly developed model of progressive rapid ventricular pacing induced heart failure in rabbits. METHODS: Six animals underwent progressive pacing with incremental rates (330 beats per min (bpm) to 380 bpm over 30 days), resulting in congestive heart failure (CHF). Five animals underwent pacing at 330 bpm for 10 days only (early LV dysfunction, ELVD) and five additional animals served as control group (CTRL). RESULTS: ELVD was characterized by decreased mean arterial pressure (P=0.05 vs. CTRL) as well as significantly impaired LV function (LV fractional shortening (FS) P<0.01 vs. CTRL) and dilatation (P<0.01 vs. CTRL). CHF was characterized by further decreased mean arterial pressure (P<0.01 vs. ELVD), further impaired LV function (FS P<0.03 vs. ELVD) and dilatation (P<0.01 vs. CTRL). In control animals, significant ANP expression was observed only in atrial tissue (P<0.02 vs. BNP) while BNP expression was ubiquitous but marginal (LV P<0.05 vs. ANP). In ELVD, activation of ANP (atria and LV P<0.05 vs. CTRL) and BNP (atria P<0.05 vs. CTRL, LV n.s.) was observed. In CHF, LV-BNP increased further markedly (P<0.01 vs. CTRL, P<0.05 vs. ELVD) while atrial ANP and BNP expression as well as LV ANP expression remained unchanged (all P=n.s. vs. ELVD). CONCLUSION: The current studies demonstrate differential activation of atrial and LV ANP and BNP under normal conditions and during the progression to heart failure and provide a molecular basis for the superiority of BNP as marker of LV dysfunction and CHF. PMID- 11476752 TI - Long-term treatment with neutral endopeptidase inhibitor improves cardiac function and reduces natriuretic peptides in rats with chronic heart failure. AB - OBJECTIVE: Increased secretion of atrial and brain natriuretic peptide (ANP and BNP) from hearts is known to exhibit favorable effects in patients and animals with heart failure, and inhibition of neutral endopeptidase (NEP), an enzyme that degrades ANP and BNP, may further increase these peptide levels. However, it is still unknown whether such elevation of the ANP and BNP may offer a therapeutic benefit to the progression of chronic heart failure (CHF). We examined the effects of ONO-9902, a novel NEP inhibitor, on changes in hemodynamic parameters, NEP activity and neurohumoral factors in rats with CHF induced by left coronary artery ligation (CAL). METHODS: Male Wistar rats (220-240 g) were subjected to induction of acute myocardial infarction by CAL. Rats were orally treated with ONO-9902 (300 mg/kg/day) from the 1st to 6th week after the operation. Hemodynamic and/or biochemical assessments were performed at the 1st and 6th weeks after the operation. RESULTS: A single administration of ONO-9902 inhibited the plasma and kidney NEP activities and thereby further augmented the elevation of plasma ANP concentration in rats with CAL at the 1st week after the operation. In rats with CAL at the 6th week after the operation, the left ventricular end diastolic pressure (LVEDP) increased and cardiac output index (COI) decreased as compared with those of sham-operated rats. These changes were accompanied by marked increases in the plasma ANP, BNP and endothelin-1 (ET-1). Chronic treatment with ONO-9902 attenuated the increase in LVEDP and the decrease in COI. These changes were associated with a decrease in plasma ANP, BNP and ET-1 concentrations. CONCLUSIONS: The results suggest that chronic treatment with NEP inhibitor improves depressed cardiac function in rats with CHF. ONO-9902 may offer a new and possible therapeutic approach in patients with CHF. PMID- 11476753 TI - The dopamine D3 receptor partial agonist, BP 897, is an antagonist at human dopamine D3 receptors and at rat somatodendritic dopamine D3 receptors. AB - Recent studies have fueled the interest in dopamine D3 receptor antagonists and partial agonist for the treatment of psychosis and drug abuse, respectively. N-[4 [4-(2-methoxyphenyl)-1-piperazinyl]butyl]naphthalene-2-carboxamide (BP 897) is a dopamine D3 receptor selective ligand recently described as partial agonist with potential effects on drug-dependence. The aim of the present study was to determine both the functional activity of BP 897 at human dopamine D3 receptors expressed in Chinese hamster ovary (CHO) cells and in an electrophysiological in vivo model of dopaminergic activity. BP 897 failed to stimulate the human dopamine D3 receptor and showed antagonistic effects (cpIC(50)=9.51) in a [(35)S]GTPgammaS binding assay in cells expressing the human dopamine D3 receptor. In vivo, BP 897 up to 8.2 mg/kg, i.v., had no agonistic effects on firing rate of substantia nigra dopaminergic neurons and antagonized the quinpirole-induced inhibition of firing (DID(50)=1.1 mg/kg). Our data demonstrate that BP 897 acts, in vivo and in vitro, as a dopamine D3 receptor antagonist. PMID- 11476754 TI - Chelerythrine and genistein inhibit the endothelin-1-induced increase in myofilament Ca(2+) sensitivity in rabbit ventricular myocytes. AB - We performed experiments to elucidate the cellular mechanism for the biphasic inotropic response to endothelin-1 of single rabbit ventricular myocytes loaded with a fluorescent dye, acetoxymethylester of indo-1. Endothelin-1 at 10 nM elicited a biphasic inotropic effect: a transient decrease in cell shortening and Ca(2+) transients followed by an increase in cell shortening without significant elevation of peak Ca(2+) transients. The selective endothelin ET(A) receptor antagonist FR139317 (2(R)-[2(R)-[2(S)-[(1-hexahydro-1H-azepinyl)]carbonyl]amino-4 methylpentanoyl]amino-3-[3-(1-methyl-1H-indolyl)propionyl]amino-3-(2 pyridyl)propionic acid) at 1 microM abolished the biphasic effect of endothelin-1 on cell shortening and Ca(2+) transients. The selective protein kinase C inhibitor chelerythrine at 1 microM and the tyrosine kinase inhibitor genistein at 5 microM inhibited the endothelin-1-induced increase in cell shortening without significantly affecting Ca(2+) transients and the transient decrease in cell shortening and Ca(2+) transients. The present results indicate that both protein kinase C and tyrosine kinase may contribute to the increase in myofilament Ca(2+) sensitivity induced by endothelin-1, whereas the decrease in Ca(2+) transients induced by endothelin-1 may be mediated by a signalling pathway different from that involved in the increase in cardiac contractility in rabbit ventricular myocytes. PMID- 11476755 TI - Inhibitory effect of endothelin-1 on the isoproterenol-induced chloride current in human cardiac myocytes. AB - It is still controversial whether the cAMP-activated Cl(-) current (I(Cl,cAMP)) is expressed in human cardiomyocytes. The whole-cell configuration of the voltage clamp technique was used to examine in detail the I(Cl,cAMP) in single human atrial and ventricular myocytes. Human cardiomyocytes were enzymatically isolated from atrial or ventricular specimens obtained from open-heart surgery or cardiac transplantation, respectively. Isoproterenol (1 microM) or forskolin (10 microM) was used to activate the cAMP second-messenger system. The isoproterenol- or forskolin-induced Cl(-) current was elicited in 12 of 54 atrial myocytes but was completely absent from ventricular myocytes. The isoproterenol-induced Cl(-) current in atrial myocytes was time-independent and had a reversal potential close to zero. Endothelin-1 (30 nM) inhibited the isoproterenol-induced Cl(-) current by 75+/-6% (n=4). This inhibitory effect of endothelin-1 was attenuated by pretreating atrial myocytes with the endothelin ET(A) receptor antagonist, BQ485, but not with the ET(B) receptor antagonist, BQ-788. The results provide evidence that the I(Cl,cAMP) exists in human atria, but not ventricle, and is inhibited by endothelin-1. PMID- 11476756 TI - Anticonvulsant activity of a mGlu(4alpha) receptor selective agonist, (1S,3R,4S) 1-aminocyclopentane-1,2,4-tricarboxylic acid. AB - The metabotropic Group III agonist, (1S,3R,4S)-1-aminocyclopentane-1,2,4 tricarboxylic acid (ACPT-1), selective for the mGlu(4alpha) receptor, suppresses sound-induced seizures in DBA/2 mice following its intracerebroventricular (i.c.v.) administration (ED(50) 5.6 [2.9-10.7], nmol i.c.v., 15 min, clonic phase) and in genetically epilepsy-prone (GEP) rats following focal administration into the inferior colliculus (ED(50) 0.08 [0.01-0.50], nmol, 60 min, clonic phase). ACPT-1 also protects against clonic seizures induced in DBA/2 mice by the Group I agonist, (RS)-3,5-dihydroxyphenylglycine (3,5-DHPG) (ED(50) 0.60 [0.29-1.2], nmol i.c.v.) and by the Group III antagonist, (RS)-alpha methylserine-O-phosphate (MSOP) (ED(50) 49.3 [37.9-64.1], nmol i.c.v.). Another Group III agonist, (RS)-4-phosphonophenyl-glycine (PPG), preferentially activating the mGlu(8) receptor, previously shown to protect against sound induced seizures in DBA/2 mice and GEP rats, also protects against seizures induced in DBA/2 by 3,5-DHPG (ED(50) 3.7 [2.4-5.7], nmol i.c.v.) and by the Group III antagonist, MSOP (ED(50) 40.2 [21.0-77.0], nmol i.c.v.). At very high doses (500 nmol i.c.v. and above), Group III antagonists have pro-convulsant and convulsant activity. The anticonvulsant protection against sound-induced seizures in DBA/2 mice provided by a fully protective dose (20 nmol, i.c.v.) of the mGlu(4) receptor agonist ACPT-1, is partially reversed by the co-administration of the Group III antagonists, MSOP, (RS)-alpha-methyl-4-phosphonophenylglycine (MPPG) or (S)-2-amino-2-methyl-4-phosphonobutanoic acid (MAP4), in the 20-50 nmol dose range. At doses of 50-200 nmol, MPPG and MAP4 cause further reversal of the ACPT-1 anticonvulsant protection, while the MSOP effect on ACPT-1 protection is abolished at higher doses. In contrast, the anticonvulsant protection against sound-induced seizures in DBA/2 mice provided by a fully protective dose (20 nmol, i.c.v.) of the mGlu(8) receptor agonist PPG, is not significantly affected by the co-administration of the same Group III antagonists, MSOP, MPPG or MAP4. We conclude that activation of either mGlu(4alpha) or mGlu(8) receptors confer anticonvulsant protection in DBA/2 mice. Furthermore, the metabotropic Group III receptor antagonists, MSOP, MPPG, and MAP4 appear to be functionally selective for the mGlu(4) receptor in this system. PMID- 11476757 TI - Tyrosine depletion attenuates the behavioural stimulant effects of amphetamine and cocaine in rats. AB - Neurochemical studies show that a tyrosine-free amino acid mixture depletes brain tyrosine and decreases dopamine synthesis and release. Here, we tested whether such a mixture would reduce the behavioural effects of amphetamine and other psychostimulants. A tyrosine-free amino acid mixture decreased the behavioural activation induced by both D-amphetamine (2 mg/kg s.c.) and cocaine (2 mg/kg s.c.). In contrast, the activation induced by the dopamine agonist, apomorphine (0.75 and 5 mg/kg s.c.), or the 5-hydroxytryptamine releasing agent, p chloroamphetamine (2 mg/kg s.c.) was not altered. These findings provide behavioural evidence that tyrosine-free amino acid mixtures reduce presynaptic dopamine function in the brain. PMID- 11476758 TI - Effects of alpha-lipoic acid on deoxycorticosterone acetate-salt-induced hypertension in rats. AB - We investigated the potential of natural occurring antioxidant alpha-lipoic acid to prevent hypertension and hypertensive tissue injury induced by deoxycorticosterone acetate (DOCA) and salt in rats. Two weeks after the start of DOCA-salt treatment, the rats were given alpha-lipoic acid (10 or 100 mg/kg/day, s.c.) or its vehicle for 2 weeks. Uninephrectomized rats without DOCA-salt treatment served as sham-operated controls. In vehicle-treated DOCA-salt rats, systolic blood pressure increased markedly after 3-4 weeks. Daily administration of 100 mg/kg alpha-lipoic acid for 2 weeks suppressed the increase in systolic blood pressure, whereas 10 mg/kg alpha-lipoic acid did not affect the progression of DOCA-salt-induced hypertension. When the degree of vascular hypertrophy of the aorta was morphometrically evaluated at 4 weeks, there were significant increases in media cross-sectional area in vehicle-treated DOCA-salt rats compared with sham-operated rats. The development of vascular hypertrophy was markedly suppressed by alpha-lipoic acid at 100 mg/kg but not at 10 mg/kg. Histopathological examination of the kidney in vehicle-treated DOCA-salt rats revealed fibrinoid-like necrosis in glomeruli and thickening of small arteries. In these animals, creatinine clearance decreased, and fractional excretion of Na(+), urinary excretion of protein and N-acetyl-beta-glucosaminidase increased. Such renal lesions and dysfunctions were ameliorated in DOCA-salt rats given alpha-lipoic acid. In addition, a marked increase in endothelin-1 content in both the aorta and kidney was evident in vehicle-treated DOCA-salt rats compared with findings in sham-operated rats. Significant attenuation of this increase occurred in alpha-lipoic acid-treated DOCA-salt rats. These results suggest that administration of alpha-lipoic acid to DOCA-salt hypertensive rats lessens the increased blood pressure and protects against renal and vascular injuries, possibly through the suppression of renal and vascular endothelin-1 overproduction. PMID- 11476759 TI - Characterization of alpha1-adrenoceptor-mediated contraction in the mouse thoracic aorta. AB - In the mouse thoracic aorta, noradrenaline, adrenaline, phenylephrine and methoxamine behaved as full agonists. The pA(2) values for 8-[2-[4-(2 methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4,5]decane-7,9-dione dihydrochloride (BMY 7378) against each agonist were in good agreement with the generally accepted affinity value of alpha(1D)-adrenoceptors. 5-Methylurapidil, 2 [2,6-dimethoxyphenoxyethyl]aminomethyl-1,4-benzodioxane hydrochloride (WB 4101) and prazosin inhibited the contraction in response to noradrenaline. A significant correlation was obtained between the antagonist affinities in mouse thoracic aorta and those of native alpha(1D)-adrenoceptors in rat thoracic aorta or with those of cloned alpha(1d)-adrenoceptors, but not with those for either alpha(1a)- or alpha(1b)-adrenoceptors. Buspirone behaved as a partial agonist in mouse thoracic aorta, the contraction of which was antagonized by BMY 7378 with a pA(2) value (8.49) consistent with that found against noradrenaline (8.43). Clonidine acted as a partial agonist (pD(2)=5.94). The pK(p) value for clonidine against noradrenaline was similar to the pD(2) value for clonidine. The apparent pK(B) value for BMY 7378 against clonidine was similar to the pA(2) value against other full agonists used in the present study. These results suggest that the alpha(1D)-adrenoceptor subtype exists, and that the full agonists and the partial agonists evoke the contraction mediated through the alpha(1D)-adrenoceptor in mouse thoracic aorta. PMID- 11476760 TI - Endothelial dysfunction induced by oxidized low-density lipoproteins in isolated mouse aorta: a comparison with apolipoprotein-E deficient mice. AB - We characterized the acute effects of oxidized low-density lipoproteins (oxidized LDL) on vascular reactivity in isolated aorta from wild-type C57BL/6J mice, and compared these with the chronic alterations in vascular function observed in apolipoprotein-E gene knockout [ApoE(-/-)] mice fed a high-fat diet, which results in hyperlipidemia and atherosclerosis. In the abdominal (but not thoracic) aorta, oxidized-LDL (100 microg/ml) reduced relaxations induced by acetylcholine (10(-9) M-10(-5) M), which are mediated entirely by nitric oxide (NO). The relaxations induced by the NO donor S-nitroso-N-acetylpenicillamine (SNAP, 10(-8) M-10(-4) M), the cyclic GMP analogue 8-bromo cyclic GMP (100 microM) and the nonspecific vasodilator papaverine (100 microM) were not changed by oxidized-LDL. Native LDL had no effect on vasorelaxations. The attenuation of endothelium-dependent relaxations caused by oxidized-LDL mimicked the endothelial dysfunction found in ApoE(-/-) mice. These results are consistent with the suggestion that oxidized-LDL has an important role in the pathogenesis of endothelial NO dysfunction associated with hyperlipidemia and atherosclerosis in these mice. PMID- 11476761 TI - Investigation of the effects of YM-31636, a novel 5-HT3 receptor agonist, on defecation in normal and constipated ferrets. AB - We examined the effects of YM-31636 (2-(1H-imidazol-4-ylmethyl)-8H-indeno[1,2 d]thiazole monofumarate), a newly synthesized 5-HT(3) receptor agonist, on defecation in normal and constipated ferrets, and evaluated it as an agent against constipation. YM-31636 facilitated defecation without inducing diarrhea or emetic episodes. This effect occurred within 1 h after oral administration, mostly within 30 min, whereas sodium picosulfate, a widely used laxative, tended to increase the frequency of defecation for several hours with much lower peak incidence than that of YM-31636, and induced diarrhea. UK14304 (brimonidine), an alpha2 receptor agonist, and morphine reduced the frequency of defecation and YM 31636 restored it. These effects of YM-31636 were antagonized by ramosetron, a 5 HT(3) receptor antagonist. These results suggest that YM-31636 could be promising in the treatment of constipation. Because of an early and reliable onset of action compared with sodium picosulfate, YM-31636 could make it easier to control the time of defecation. PMID- 11476762 TI - Release of nerve growth factor by human pulmonary epithelial cells: role in airway inflammatory diseases. AB - Elevated levels of nerve growth factor (NGF) have been detected in the bronchoalveolar lavage fluid of patients with asthma. However, the source of this enhanced mediator production is not known. Here, we investigate the production of NGF from a human airway epithelial cell line (A549). Under basal conditions, A549 cells generated NGF in a time-dependent fashion. However, basal release was significantly augmented in a concentration-dependent manner in cells treated with interleukin-1beta (IL-1beta) or tumour necrosis factor-alpha (TNF-alpha) and inhibited by dexamethasone. These data suggest that NGF released from structural cells may be an important target for the anti-inflammatory effects of steroids in asthma therapy. PMID- 11476763 TI - Glibenclamide's action in the hypothalamus alters peripheral glucose homeostasis. AB - Bilateral injections of 0.2 nmol of glibenclamide, targeted at the ventromedial hypothalamus of albino rats, significantly impaired recovery of blood glucose levels following insulin-induced hypoglycemia (P=0.0026). Similar injections of 0.2 and 2.0 nmol glibenclamide reduced blood glucose by 23.6 and 40.8 mg%, respectively, in otherwise untreated rats (P<0.01 and P<0.001). Intravenous injection of these doses of the drug did not lower blood glucose. The results support the hypothesis that K(ATP)(+) channels within the hypothalamus serve a counterregulatory function in the maintenance of peripheral glucose levels. PMID- 11476764 TI - Treating endometriosis as an autoimmune disease. AB - OBJECTIVE: To review the literature on the role of autoimmunity in the etiology of endometriosis, compare the similarities in the pathophysiologies between endometriosis and autoimmune diseases, and discuss the use of immunomodulators currently used to treat autoimmune diseases as potential therapies for endometriosis. DESIGN: The literature on endometriosis and other autoimmune diseases was reviewed, and summary data are presented. RESULTS: Endometriosis shares many similarities with autoimmune diseases such as rheumatoid arthritis, Crohn's disease, and psoriasis. These similarities include elevated levels of cytokines, decreased cell apoptosis, and T- and B-cell abnormalities. Because the use of immunomodulators and inflammatory modulators has proven to be an effective means of medical management for these autoimmune diseases, similar therapies may prove useful in treating endometriosis. CONCLUSION(S): Although substantial evidence indicates that endometriosis at least shares many similarities with autoimmune diseases, endometriosis is primarily treated by using compounds that induce a hypoestrogenic environment. A review of the literature combined with the shortcomings of current means of medical management for endometriosis support the postulate that treatment of endometriosis with immunomodulators and inflammatory modulators is warranted. PMID- 11476765 TI - Human reproductive cloning: not ready for prime time. PMID- 11476766 TI - Effects of androgen supplementation of hormone replacement therapy on the vascular reactivity of cerebral arteries. AB - OBJECTIVE: To determine the effect of the androgen supplementation of hormone replacement therapy (HRT) on the vascular reactivity of cerebral arteries. DESIGN: Open randomized study. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Forty postmenopausal women who were treated with sequential HRT (transdermal E2 50 microg/d + medroxyprogesterone acetate 10 mg/d for 12 days every other month) for > or =1 year and < or =5 years. INTERVENTION(S): Testosterone undecanoate (40 mg/d, p.o.) was randomly administered to 20 patients during ongoing HRT; the other 20 served as controls. Doppler evaluations of the internal carotid and middle cerebral arteries were performed together with lipid levels assessments. A visual analogue scale (VAS) was used to evaluate various parameters relating to sexual life and well-being. MAIN OUTCOME MEASURE(S): Pulsatility index (PI) of the arteries, VAS assessment of psychophysical well-being. RESULT(S): The administration of testosterone undecanoate during HRT induced an increase in the PI of the middle cerebral artery and a reduction of high-density lipoprotein cholesterol. Sexual desire and satisfaction were greatly improved. CONCLUSION(S): In postmenopausal women, androgen supplementation during HRT can partially counteract the beneficial effects of estrogens on cerebral vascular reactivity and lipid profiles, but sexual desire and satisfaction are greatly improved. PMID- 11476767 TI - Oral dehydroepiandrosterone supplementation modulates spontaneous and growth hormone-releasing hormone-induced growth hormone and insulin-like growth factor-1 secretion in early and late postmenopausal women. AB - OBJECTIVE: To evaluate the effects of dehydroepiandrosterone (DHEA) supplementation on the growth hormone-releasing hormone-growth hormone (GHRH-GH) axis in lean and obese postmenopausal women. DESIGN: Prospective study. SETTING: Postmenopausal women in a clinical research environment. PATIENT(S): Thirty-one postmenopausal women were divided in two groups by age (50 to 55 and 60 to 65 years). Within each group, lean and obese patients were considered. INTERVENTION(S): All patients underwent hormonal evaluations before and at the third and sixth month of therapy (50 mg of DHEA orally each day) and a GHRH test (1 microg/kg) before and at the sixth month of treatment. Ultrasound and bone mass density (BMD) examinations were performed before and after the sixth month of therapy. MAIN OUTCOME MEASURE(S): Plasma dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), E1, E2, androstenedione (A), testosterone (T), osteocalcin, GH, insulin-like growth factor 1 (IGF-1) concentrations. RESULT(S): The levels of all of the steroids that derived from DHEA metabolism (E1, E2, A, T, DHEAS) and osteocalcin were increased in plasma under DHEA supplementation. The supplementation protocol also increased the levels of GH and IGF-1. However, GHRH-induced GH and IGF-1 responses were not modified by DHEA supplementation. CONCLUSION(S): Administration of DHEA significantly affects several endocrine parameters in early and late postmenopausal women independently from body mass index. Our data support the hypothesis that DHEA treatment acts similarly to estrogen-progestin replacement therapy on the GHRH-GH-IGF-1 axis. This suggests that DHEA is more than a more than a simple "diet supplement" or "antiaging product"; rather it should be considered an effective hormonal replacement treatment. PMID- 11476768 TI - A prospective controlled study of karyotyping for 430 consecutive babies conceived through intracytoplasmic sperm injection. AB - OBJECTIVE: To compare the karyotype of babies conceived through ICSI with that of naturally conceived babies. DESIGN: Prospective controlled study. SETTING: The Egyptian IVF-ET Center, Cairo, Egypt. PATIENT(S): Four hundred and thirty babies conceived through ICSI and 430 babies conceived naturally. INTERVENTION(S): ICSI and karyotyping. MAIN OUTCOME MEASURE(S): Abnormal karyotype. RESULT(S): Four hundred and thirty consecutive babies conceived through ICSI who were delivered in one hospital had 15 abnormal karyotypes (3.5%). Of the 15 babies, 7 were of female phenotype and 8 of male phenotype. Six babies had sex chromosome anomalies, 8 had autosomal anomalies, and 1 had combined sex chromosome and autosomal anomalies. A control group of 430 consecutive babies conceived naturally who were delivered in one hospital had no abnormal karyotype. The difference between the two groups was significant (P<.001). CONCLUSION(S): ICSI carries a small but significant increased risk of abnormal karyotyping to the offspring. This risk appears to be equally distributed between autosomal and sex chromosome anomalies. PMID- 11476769 TI - Thyroid hormones and thyroid antibodies in infertile males. AB - OBJECTIVE: To investigate the incidence of thyroid dysfunction, thyroid antibodies, and the correlation with semen and hormonal parameters in infertile men. DESIGN: Prospective study. SETTING: University-based andrology laboratory. PATIENT(S): Three hundred five infertile men with idiopathic infertility. INTERVENTION(S): Medical history, clinical examination, semen analysis, measurement of free thyroxin (fT4), free triiodothyronine (fT3), basal thyroid stimulating hormone (bTSH), LH, FSH, T, free testosterone (fT), PRL, E2, sex hormone-binding globulin (SHBG), DHEAS, and the thyroid antibodies thyreoglobulin antibody (TGA), thyroid peroxidase antibody (TPO-Ab), and thyroid receptor antibody (TRAK). MAIN OUTCOME MEASURE(S): Incidence of thyroid dysfunction and thyroid antibodies, as well as the correlation with hormones and the results of semen analyses. RESULT(S): No manifest thyroid dysfunction was observed. Latent thyroid dysfunction and latent hypothyroidism were diagnosed in 11.5% and 3% of infertile men, respectively. No correlation between thyroid dysfunction and semen parameters was detected. bTSH correlated significantly with PRL (P<.001). Thyroid antibodies were elevated in 7.5%. Elevated TPO-Ab were significantly correlated with pathozoospermia (P=.036) and asthenozoospermia (P=.049). CONCLUSION(S): Latent thyroid dysfunction had no impact on semen parameters. In patients with elevated TPO-Ab levels, pathozoospermia or asthenozoospermia should be considered. PMID- 11476770 TI - Members of the Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway are present and active in human sperm. AB - OBJECTIVE: To investigate whether components of the Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway are present and active in human sperm. DESIGN: Comparative study. SETTING: Reproductive biology department. PATIENT(S): Nine sperm donors. INTERVENTION(S): Sperm were exposed to interferon-alpha (IFN-alpha), IFN-gamma, interleukin-12 (IL-12), Ca2+ ionophore (A23187), or progesterone under capacitating conditions. MAIN OUTCOME MEASURE(S): Cell lysates prepared from sperm and Jurkat T-cell line were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the expression of JAKs (1 3 and TYK 2) and STATs (1-6) was examined by Western blot analysis. Effect of IFN alpha, IFN-gamma, IL-12, A23187, or progesterone on sperm STAT 1 or STAT 4 phosphorylation was determined by phospho-STAT 1 antibody or antiphosphotyrosine (APT) Western blot analysis. Indirect immunofluorescence and confocal laser scanning microscopy was used to confirm the specific staining of anti-TYK 2, anti STAT 1, and anti-STAT 4 antibodies. RESULT(S): By Western blot analysis, only antibodies to TYK 2 of the JAK family, and antibodies to STAT 1 and STAT 4 members of the STAT family specifically recognized protein bands corresponding to TYK 2, STAT 1, and STAT 4 described in other cell types. By confocal microscopy, antibodies to TYK 2 reacted with the sperm tail as well as the apical region of sperm head, whereas antibodies to STAT 1 and STAT 4 reacted with the apical region of the sperm head. Tyrosine phosphorylation of STAT 1 in capacitated sperm was enhanced by IFN-alpha and IFN-gamma, and that of STAT 4 was enhanced by IL 12. Both A23187 and progesterone markedly inhibited tyrosine phosphorylation of sperm STAT 4. CONCLUSION(S): Members of the JAK/STAT proteins, TYK 2, STAT 1, and STAT 4 are present and active in human sperm. The localization of STAT 1 and STAT 4 proteins to the apical region of the sperm head and their activation by IFN alpha, IFN-gamma, or IL-12 implicate a role for sperm STAT proteins in fertilization. We hypothesize that sperm-derived phosphorylated STAT 1 and STAT 4 could contribute to the pool of transcription factors during sperm-oocyte fusion as well as transmit signal to the oocyte nucleus. Therefore, defects in sperm TYK 2 and STAT 1- or STAT 4-mediated signaling pathway may have relevance to male factor infertility. PMID- 11476771 TI - Molecular evaluation of two major human sperm fibrous sheath proteins, pro hAKAP82 and hAKAP82, in stump tail sperm. AB - OBJECTIVE: To determine whether mutations in the pro-hAKAP82 gene and the resulting pro-hAKAP82 and hAKAP82 proteins were associated with the infertility seen in a patient with stump tail sperm. DESIGN: Case report. SETTING: Academic research and teaching environment, tertiary care hospital. PATIENT(S): A single, infertile Caucasian male diagnosed with essentially 100% stump tail sperm. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Electrophoresis, silver staining, and immunoblotting of patient and control sperm proteins; RII (type II regulatory subunit of protein kinase-A) overlay assay of patient and control sperm proteins, partial DNA sequence analysis of patient's pro-hAKAP82 gene; indirect immunofluorescence and immunogold electron microscopy of patient and control sperm. RESULTS(S): No significant abnormalities in the size or amount of pro hAKAP82 and hAKAP82 or in the ability of these proteins to bind the regulatory subunit of protein kinase-A were identified in the patient's sperm. Partial sequence analysis of the patient's pro-hAKAP82 gene was identical to the published normal sequence. Indirect immunofluorescence and immunoelectron microscopy of sperm localized pro-hAKAP82/hAKAP82 to the sperm flagellum and demonstrated that the proteins were present in a disorganized, amorphous region, which apparently represented the fibrous sheath. CONCLUSION(S): These results suggest that, although pro-hAKAP82 and hAKAP82 localize to the correct structural component of the flagellum and are not directly responsible for the stump tail phenotype, they are unable to assemble normally into the fibrous sheath. Although this study did not identify abnormalities in the pro-hAKAP82 gene or its resulting proteins in a patient with stump tail sperm, several regions of the gene and protein remain to be examined. PMID- 11476772 TI - Influence of antisperm antibodies on the acrosome reaction as determined by flow cytometry. AB - OBJECTIVE: To evaluate the influence of antisperm antibodies on the acrosome reaction (AR). DESIGN: Clinical study. SETTING: University of Marburg, Department of Andrology, Clinical Training Center of the European Academy of Andrology. PATIENT(S): Spermatozoa from a pool of healthy donors were incubated with 30 seminal plasma samples from infertile men containing antisperm antibodies; they were compared to a control group of 10 samples without antisperm antibodies and five samples with buffer only. INTERVENTION(S): The spontaneous acrosome reaction (SAR) and the induced acrosome reaction (IAR) by calcium ionophore A23187 were observed and determined by means of a flow cytometer. Flow cytometric double staining estimates of acrosomal integrity were determined by using a monoclonal antibody (TUS 19), marked with a secondary fluorescein isothiocyanate-labeled antibody. Cell viability was determined by counterstaining with propidium iodide (PI). MAIN OUTCOME MEASURE(S): Number of acrosome reacted spermatozoa. RESULT(S): The spermatozoa treated with antisperm antibodies showed significantly higher SAR and IAR responses than the control group. CONCLUSION(S): Some of the antisperm antibodies from individual patients are able to enhance the acrosome reaction in donor sperm, but none of them appeared to inhibit acrosome reaction. PMID- 11476773 TI - Embryo fragmentation in vitro and its impact on treatment and pregnancy outcome. AB - OBJECTIVE: To determine the impact of embryo fragmentation on pregnancy, obstetric, and perinatal outcome. DESIGN: Retrospective analysis of embryo transfers that were homogeneous in regard to the degree of fragmentation. SETTING: Fertility center. PATIENT(S): A cohort of 460 fresh embryo transfers. INTERVENTION(S): A total of 164 pregnancies were analyzed for the incidence of antepartum complications during gestation, obstetric (multiple pregnancy, preterm delivery, cesarean section), and perinatal outcome (sex, birth weight, admission to neonatal intensive care unit, malformations). MAIN OUTCOME MEASURE(S): Implantation and clinical pregnancy rate, obstetric and perinatal outcome. RESULT(S): Embryo fragmentation and number of embryos per transfer showed a significant influence on clinical pregnancy and implantation rate. No such relation was found concerning complications, multiple pregnancy rate, incidence of cesarean section, gestation week, birth weight, and average time at the neonatology. On the other hand, pregnancies derived from bad-quality embryos had a significantly higher rate of malformations. CONCLUSION(S): The higher percentage of malformations found in bad-quality embryos may be due to a higher percentage of apoptotic features and chromosomal disorders. For ethical reasons, the transfer of embryos with >50% fragmentation should be considered only after consultation with the patient. PMID- 11476774 TI - Is tubal embryo transfer of any value? A meta-analysis and comparison with the Society for Assisted Reproductive Technology database. AB - OBJECTIVE: To analyze published randomized trials of ZIFT and ET via meta analysis and compare the results with those of the Society for Assisted Reproductive Technology database. DESIGN: Meta-analysis and comparison to SART data sets for 1991-1996. SETTING: University medical center. PATIENT(S): Patients from the literature with infertility, randomized to either tubal or uterine embryo transfer. INTERVENTION(S): All published articles in English were identified using an electronic database spanning January 1966 to December 1998 by keyword and text word searches, supplemented with a hand search through the references of original studies, review articles, and conference abstracts to identify randomized trials comparing ZIFT and IVF-ET. Additional data was obtained through correspondence with authors. MAIN OUTCOME MEASURE(S): Implantation and clinical pregnancy rates were compared. Ectopic pregnancy rate was a secondary outcome measure. RESULT(S): Six randomized controlled trials including 548 cycles, 514 retrievals, and 388 transfers were reviewed. Demographic and stimulation and transfer details were comparable between the groups. Implantation and pregnancy rates did not differ significantly, and there was a trend toward increased risk of ectopic pregnancy with ZIFT. CONCLUSION(S): Published randomized trials suggest that there is no difference in implantation and pregnancy rates between women undergoing ZIFT and IVF-ET. PMID- 11476775 TI - Increased progesterone/estradiol ratio in the late follicular phase could be related to low ovarian reserve in in vitro fertilization-embryo transfer cycles with a long gonadotropin-releasing hormone agonist. AB - OBJECTIVE: To gain insight into the physiologic as well as the clinical significance of premature luteinization in the long gonadotropin-releasing hormone agonist (GnRH-a) cycles and to evaluate whether it may be a manifestation of low ovarian reserve. DESIGN: Prospective evaluation. SETTING: A university affiliated reproductive medicine unit. PATIENT(S): Seventy-six consecutive infertile women. INTERVENTION(S): The long GnRH-a protocol was used for IVF-ET treatment. MAIN OUTCOME MEASURE(S): Women in the study were prospectively evaluated in their first cycle of treatment and were divided into those with (study group) or without premature luteinization (control group). Premature luteinization was defined as P/E2 ratio of more than 1 on the day of hCG administration. RESULTS(S): Thirty-one (41%) of the women in the study demonstrated premature luteinization. Patients' characteristics were comparable between the two groups. Late follicular P/E2 ratio was significantly and considerably higher in the study as compared to the control group, 2.4 +/- 1.7 and 0.7 +/- 0.2, respectively. Ovarian reserve parameters including day 3 FSH, E2 level on hCG day, total amount of hMG, number of follicles, oocytes, and embryos were significantly inferior in the study as compared to the control group. P levels on hCG day were significantly higher in the study as compared to the control group, 1.9 +/- 0.7 ng/mL and 1.2 +/- 0.6 ng/mL, respectively. However, LH levels on hCG day did not differ between the groups, 1.4 +/- 0.7 mIU/mL and 1.2 +/- 0.7 mIU/mL, respectively. The clinical pregnancy rate was significantly lower in the premature luteinization group as opposed to controls, 13% and 42%, respectively. CONCLUSION(S): Premature luteinization, defined as late follicular P/E2 >1, in long GnRH-a cycles seems to adversely affect clinical outcome. Our findings in this setting support the notion that premature luteinization could be related to low ovarian reserve and that this manifestation is not necessarily an LH-dependent event. PMID- 11476776 TI - Can we abandon routine evaluation of serum estradiol levels during controlled ovarian hyperstimulation for assisted reproduction? AB - OBJECTIVE: To assess whether abandoning measurement of serum estradiol (E2) and spacing ultrasound evaluations at greater intervals had an effect on the results of assisted reproduction technology (ART). DESIGN: A retrospective comparison of two consecutive periods. SETTING: Division of Assisted Reproduction Technology, Department of Obstetrics and Gynecology, Ha'Emek Medical Center, Afula, Israel. PATIENT(S): One thousand nine hundred and eighty-five controlled ovarian hyperstimulation (COH) cycles for ART were initiated during the years 1996 to 1999. INTERVENTION(S): During the first 2 years an intensive follow-up protocol was used that included E2 blood levels measurements. In the next 2 years a less intensive protocol was adopted that did not use E2 measurements. MAIN OUTCOME MEASURE(S): ART results and the rate of ovarian hyperstimulation syndrome (OHSS). RESULT(S): The patients' background characteristics did not differ between the two periods. The cancellation rate was not different (9.8% vs. 7.2%). There was no difference in the duration of stimulation or the amount of gonadotropins used. The number of oocytes retrieved (12.1 +/- 9.3 vs. 9.6 +/- 6.3), fertilization rates (74% vs. 75%), and clinical pregnancy rates (26.2% vs. 27.9%) did not differ. The incidence of severe ovarian hyperstimulation syndrome was not significantly different between the two periods. CONCLUSION(S): Controlled ovarian hyperstimulation for ART can be done reliably without routine, serial serum E2 measurements without compromising the treatment results. PMID- 11476777 TI - Treatment of menorrhagia with the levonorgestrel intrauterine system versus endometrial resection. AB - OBJECTIVE: Treatment of menorrhagia with levonorgestrel intrauterine system (LNG IUS) and transcervical resection. DESIGN: An open, therapeutic, randomized study. SETTING: Central county hospital specializing in hysteroscopy. PATIENT(S): Two parallel groups of 30 subjects each. INTERVENTION(S): Thirty patients had a LNG IUS inserted within the first 7 days of menses; 29 patients underwent endometrial resection. MAIN OUTCOME MEASURE(S): A 12-month follow-up of menstrual blood loss and adverse events were evaluated. RESULT(S): LNG IUS group: 13 patients reported one or more pelvic adverse events, bleeding disorders (n = 6), abdominal pain (n = 4), breast tenderness (n = 3), headache, acne (n = 2), and mood changes (n = 1). Six patients discontinued treatment because of irregular bleeding (n = 3), pain (n = 2), and acne (n = 1). In both groups, general feeling of genital health increased with Visual Analogue Scale score. Nine patients reported adverse events. This included pelvic pain indicating inflammation (n = 4), bleeding (n = 3), vaginitis (n = 1), and ulceration (n = 1). Treatment success at 12 months was achieved in 20 (67%) of the 30 patients in the LNG IUS group and in 26 (90%) of the 29 patients in the transcervical resection group. Adverse events were more often reported in the LNG IUS group. CONCLUSION(S): Both treatments effectively reduced the menstrual blood loss. Furthermore, the LNG IUS treatment is reversible and has no operative hazards. PMID- 11476778 TI - Psychological correlates of functional hypothalamic amenorrhea. AB - OBJECTIVE: To determine whether mood, attitudes, or symptoms of disordered eating discriminated women with functional hypothalamic amenorrhea (FHA) from those with organic causes of amenorrhea and eumenorrhea. DESIGN: Cross-sectional comparison of women with FHA, women with organic amenorrhea, and eumenorrheic control women. SETTING: Clinical research center in an academic medical institution. PATIENT(S): Seventy-seven women > or =18 years old with time since menarche > or =5 and < or =25 years were recruited by advertisement. INTERVENTION(S): Ovulation was confirmed in eumenorrheic control women. Causes of anovulation were carefully documented in amenorrheic participants and LH pulse profiles were obtained to document the diagnosis of FHA. All participants were interviewed and completed questionnaires. MAIN OUTCOME MEASURE(S): Self-report measures of dysfunctional attitudes, coping styles, and symptoms of depression and eating disorders. RESULT(S): Women with FHA reported more depressive symptoms and dysfunctional attitudes than did eumenorrheic women, but not significantly more than women with organic amenorrhea. However, women with FHA reported significantly more symptoms of disordered eating than did either anovulatory or ovulatory women. CONCLUSION(S): The findings are consistent with the hypothesis that FHA is precipitated by a combination of psychosocial stressors and metabolic challenge. PMID- 11476779 TI - Lactational amenorrhea/anovulation and some of their determinants: a comparison of well-nourished and undernourished women. AB - OBJECTIVE: To study the effect of maternal nutritional status and some other possible determinants on lactational amenorrhea/anovulation. DESIGN: Prospective matched-pairs study. SETTING: Postpartum wards and community and academic settings. PATIENT(S): Thirty matched pairs of otherwise healthy, well-nourished (body mass index > or = 26.00 kg/m2) and undernourished (body mass index < or = 19.00 kg/m2) postpartum women were selected. INTERVENTION(S): Infant feeding pattern was recorded weekly, and infant weights, maternal body mass index, and maternal PRL levels were estimated every 4 weeks until resumption of menstruation. Ovulatory activity was determined using urinary estrone and pregnanediol glucuronide concentrations. MAIN OUTCOME MEASURE(S): Time of resumption of menstruation, regular and ovulatory. RESULT(S): Well-nourished women resumed regular menstruation significantly earlier than undernourished women but resumed ovulatory menstruation at almost the same time. Undernourished women had fewer anovulatory cycles preceding first postpartum ovulation and a higher prevalence of formula feeding. Effect of body mass index on lactational amenorrhea became nonsignificant when nonintroduction of formula feeds, maternal age, and socioeconomic status were controlled for. CONCLUSION(S): Improved maternal nutritional status has no significant effect on fertility: ovulation is not advanced despite early resumption of regular menstruation. PMID- 11476780 TI - A new ultrasound criterion for the diagnosis of polycystic ovary syndrome: the ovarian stroma/total area ratio. AB - OBJECTIVE: To evaluate whether some ultrasound parameters of ovarian morphology can discriminate between control women and patients with polycystic ovary syndrome (PCOS). DESIGN: Retrospective data analysis. SETTING: Volunteers women in an academic research environment. PATIENT(S): Eighty amenorrheic or oligomenorrheic women and 30 normal ovulatory control participants. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): We evaluated ovarian volume, area, stroma, and the stroma/total area (S/A) ratio by use of transvaginal pelvic ultrasound; and we assayed serum levels of gonadotropin, androgen, and estradiol during the early follicular phase (days 2 to 5) of the menstrual cycle in regularly cycling controls and on a random day in amenorrheic patients. RESULT(S): Patients with PCOS showed significantly higher ovarian volume, area, stroma, and mean S/A ratio when compared to multifollicular and control groups. Cut-off values have been defined for ovarian volume (13.21 mL), area (7.00 cm2), stroma (1.95 cm2), and S/A ratio (0.34). The sensitivity for PCOS diagnosis was 21%, 4%, 62%, and 100%, respectively. The S/A ratio showed the most significant correlation with the androgen levels. CONCLUSION(S): The evaluation of the S/A ratio can differentiate between PCOS and control or multifollicular women with both a sensitivity and a specificity of 100%. Furthermore, this ultrasound parameter is strictly related to hormonal milieu and to anthropometric characteristics. PMID- 11476781 TI - Absence of corpus luteum rescue by chorionic gonadotropin in women immunized with a contraceptive vaccine. AB - OBJECTIVE: To determine whether antibodies induced by a betahCG vaccine in women are competent to neutralize the luteotropic action of hCG. DESIGN: Prospective clinical study. SETTING: Hospitals and a laboratory at an academic center. PATIENT(S): Six immunized and three control women of reproductive age, participating in a clinical testing of betahCG contraceptive vaccine. INTERVENTION(S): Increasing doses of hCG simulating early pregnancy. MAIN OUTCOME MEASURE(S): Anti-hCG antibody titers, serum progesterone and urinary hCG levels, and onset of menses. RESULT(S): Administration of hCG to mimic early pregnancy sustained serum progesterone concentrations and extended the luteal phase in control women. In contrast, serum progesterone levels declined and the luteal phase was not extended if prevailing antibody titers were > or =40 ng/mL in women who had been immunized with a betahCG based vaccine. No booster effect was seen in anti-hCG titers after hCG challenge. CONCLUSION(S): Antibodies elicited by a betahCG vaccine inactivate hCG and prevent the hormone from rescuing corpus luteum, resulting in progesterone fall and normal menses. Lack of booster in the antibody response confirms the reversibility of the approach. PMID- 11476782 TI - Characteristics of uterine peristalsis in spontaneous and induced cycles. AB - OBJECTIVE: To characterize the dynamics of the intrauterine fluid-wall interface (IUFWI) from in vivo transvaginal ultrasound images by new techniques of image processing of sagittal cross-sections of the uterus, in healthy women with normal cycles and patients treated with clomiphen citrate (CC). DESIGN: Clinical study. SETTING: An ultrasound unit in a large university-affiliated municipal hospital. PATIENT(S): Twenty-five patients with normal spontaneous cycles (group A) and nine patients treated by CC (group B). INTERVENTION(S): Transvaginal ultrasound examinations. MAIN OUTCOME MEASURE(S): Transvaginal ultrasound images were processed to carry out a computational analysis of the resultant IUFWI. RESULT(S): The amplitude and the width of the IUFWI were higher in group B, but the ratio of these measurements was similar in both groups. The frequency of wall motility of group A was lower and its pattern was more symmetrical than that of group B. CONCLUSION(S): The differences in the dynamic characteristics of spontaneous and CC-induced cycles may constitute an additional parameter that should be considered in embryo transport. PMID- 11476783 TI - Salpingographic demonstration of "cobblestone" mucosa of the distal tubes is indicative of irreversible mucosal damage. AB - OBJECTIVE: To establish the predictive value of the observation of a "cobblestone" pattern of the distal tube mucosa shown on the selective salpingogram as an indicator of significant and probably irreparable damage to the ciliated epithelium. DESIGN: Clinical study. SETTING: Fertility and Laser Center and Academic Radiology Department. PATIENT(S): Patients with primary or secondary infertility and obstruction of the fallopian tubes documented on hysterosalpingogram. INTERVENTION(S): Transcervical recanalization of obstructed tubes followed by reassessment by selective salpingogram. MAIN OUTCOME MEASURE(S): Progression of mucosal disease with resultant low likelihood of intrauterine pregnancies and high probability of ectopic pregnancy. RESULT(S): In 32 asymptomatic patients, there was progression to coalescence and agglutination of mucosal folds in 13, to a thick wall pyosalpinx in 4. Five thin wall and 8 thick wall hydrosalpinges in asymptomatic patients showed no significant progression of mucosal disease but increasing peritubular fibrosis. In 12 symptomatic patients, progressive fusion of mucosal folds with polypoid hyperplasia was observed in 11, a thin wall hydrosalpinx in 2. CONCLUSION(S): Cobblestone appearance of the distal tubes heralds significant mucosal damage, prone to progressive disease and hence, a poor chance for conception. PMID- 11476784 TI - Preimplantation testing for phenylketonuria. AB - OBJECTIVE: To use preimplantation genetic diagnosis to achieve a phenylketonuria free pregnancy in a couple at 50% risk for producing an affected child. DESIGN: DNA analysis of the first and second polar bodies (PB1 and PB2) obtained from oocytes of a heterozygous mother in IVF-ET, with the goal of identifying and transferring back to the patient the embryos resulting from mutation-free oocytes. SETTING: IVF program of Reproductive Genetics Institute, Chicago, Illinois. PATIENT(S): A mother carrying the R408W mutation and a father with compound heterozygosity for R408 and Y414C mutations in phenylalanine hydroxylase (PAH) gene. INTERVENTION(S): Removal and testing for maternal mutation in PB1 and PB2 from each oocyte after standard IVF. MAIN OUTCOME MEASURE(S): DNA analysis of PB1 and PB2 indicating whether corresponding oocytes were mutation-free, for the purposes of transferring only unaffected embryos resulting from these oocytes. RESULT(S): Of 11 zygotes with both PB1 and PB2, 6 were predicted to be free of phenylketonuria. Of these, 4 were transferred, resulting in an unaffected twin pregnancy and birth of two healthy children. CONCLUSION(S): Preimplantation genetic diagnosis of phenylketonuria resulted in the birth of phenylketonuria free children. Preimplantation genetic diagnosis by PB analysis in couples with a compound heterozygous male partner is clinically useful. PMID- 11476785 TI - Evaluation of the uterine cavity with magnetic resonance imaging, transvaginal sonography, hysterosonographic examination, and diagnostic hysteroscopy. AB - OBJECTIVE: To evaluate and compare the diagnostic accuracy of magnetic resonance imaging (MRI), transvaginal ultrasonography (TVS), hysterosonographic examination (HSE), and hysteroscopy in the evaluation of the uterine cavity. DESIGN: Independent double-blind study. SETTING: University medical hospital. PATIENT(S): One hundred six consecutive premenopausal women who underwent hysterectomy for benign diseases. INTERVENTION(S): Results of MRI, TVS, HSE, and hysteroscopy were compared with the results of histopathologic examination at hysterectomy (the gold standard). RESULT(S): The overall sensitivity was MRI 0.76, TVS 0.69, HSE 0.83, and hysteroscopy 0.84. The specificity was MRI 0.92, TVS 0.83, HSE 0.90, and hysteroscopy 0.88 (MRI, HSE, hysteroscopy vs. TVS <0.05). Polyps were missed in 9 of 12 cases at MRI, 7 at TVS, 4 at HSE, and 2 at hysteroscopy (MRI vs. hysteroscopy, and TVS vs. hysteroscopy <0.05). The sensitivity for identification of submucous myomas was MRI 1.0, TVS 0.83, HSE 0.90, and hysteroscopy 0.82; the specificity was MRI 0.91, TVS 0.90, HSE 0.89, and hysteroscopy 0.87 (MRI vs. TVS, and MRI vs. hysteroscopy). Magnetic resonance imaging was significantly more precise than TVS, HSE, and hysteroscopy in determining submucous myoma in-growth (2-way ANOVA <0.05). CONCLUSION(S): For exclusion of abnormalities in the uterine cavity, MRI, HSE, and hysteroscopy were equally effective and slightly superior to TVS. Magnetic resonance imaging and TVS missed endometrial abnormalities such as polyps, but MRI and HSE were most accurate for the evaluation of submucous myomas, and MRI was superior in evaluation of exact submucous myoma in-growth. PMID- 11476786 TI - Laparoscopic treatment of complete obliteration of the cul-de-sac associated with endometriosis: long-term follow-up of en bloc resection. AB - OBJECTIVE: To evaluate symptom relief following a laparoscopic technique designed for treatment of complete obliteration of the cul-de-sac associated with endometriosis, with fertility preserved. DESIGN: Preoperative and postoperative questionnaire study of a cohort of patients with complete obliteration of the cul de-sac undergoing a standardized laparoscopic surgical treatment. SETTING: American tertiary referral center for the surgical treatment of endometriosis. PATIENTS: Eighty-four consecutive patients undergoing laparoscopic treatment of endometriosis with complete cul-de-sac obliteration with 67 replying to a postoperative questionnaire. INTERVENTIONS: Laparoscopic excision of all endometriosis including treatment of complete obliteration of the cul-de-sac by en bloc resection and bowel resection as needed. MAIN OUTCOME MEASURES: Symptom relief as measured on a 5-point ranked ordinal scale administered before and after surgery, as well as perioperative complications, postoperative fertility, and prognostic value of preoperative findings on pelvic examination. RESULTS: Symptom reduction was obtained for all symptoms related to cul-de-sac disease, particularly for patients with severe or debilitating symptoms preoperatively. There was no significant complication, and the postoperative fertility rate was 43%. Seventy-three percent of patients with obliteration of the cul-de-sac had histologically proved rectal endometriosis. Nodularity and tenderness on examination were predictive of symptom improvement. CONCLUSIONS: Aggressive laparoscopic excision of endometriosis carried out in a specialist center offers good symptom relief, especially for those with severe or debilitating symptoms. To ensure complete removal of all disease, intestinal surgery is required in most patients with complete obliteration of the cul-de-sac. PMID- 11476787 TI - Reproductive outcome after laparoscopic instillation of hyperosmolar glucose into unruptured tubal pregnancies. AB - OBJECTIVE: To determine intrauterine and ectopic pregnancy rates after local instillation of 50% glucose into unruptured tubal pregnancies. DESIGN: Retrospective cohort of 183 patients, who were observed for 16 to 108 months (median, 64 months). SETTING: University hospital. PATIENT(S): One hundred eighty three women who underwent laparoscopic instillation of hyperosmolar glucose for unruptured tubal pregnancies. INTERVENTION(S): Completion of questionnaires. MAIN OUTCOME MEASURE(S): Pregnancy rate, tubal recurrence rate. RESULT(S): A total of 124 women returned the questionnaire and had a desire for spontaneous conception. The conception rate and the intrauterine pregnancy rate were 79% and 73%, respectively. Seventy percent of all women with desire for spontaneous conception had live births. The rate of subsequent ectopic pregnancies was 12%. Tubal patency of the treated tube was demonstrated in 69% of 39 women at hysterosalpingography. CONCLUSION(S): The long-term prognosis for conception after laparoscopic instillation of hyperosmolar glucose for unruptured tubal pregnancy is favorable and comparable to that of other conservative treatments. PMID- 11476788 TI - Determinants of pregnancy rate and obstetric outcome after laparoscopic myomectomy for infertility. AB - OBJECTIVE: To determine the effect of myomectomy on infertility and to assess the factors influencing reproductive outcome. DESIGN: Retrospective study. SETTING: Tertiary care center. PATIENT(S): One hundred and three infertile women with uterine leiomyoma who had had infertility >2 years and a follow-up time >12 months were enrolled. Follow-up was complete for 88 patients, including 28 (31.8%) with primary infertility and 44 (50%) with unexplained infertility. The mean (+/-SD) age of the patients was 36.1 +/- 2.1 years. INTERVENTION(S): Laparoscopic myomectomy. MAIN OUTCOME MEASURE(S): Pregnancy rate according to patient and fibroid characteristics. RESULT(S): Forty-two patients became pregnant (40.7%). The mean (+/-SD) delay in conception was 7.5 +/- 2.6 months. Nearly 80% of the women conceived spontaneously. Of 44 pregnancies in 42 women, 36 live newborns were delivered. No dehiscence of uterine scar occurred. The pregnancy rate was significantly higher in women <35 years of age or <3 years of infertility. Women with unexplained infertility had higher pregnancy rate than did women with multifactorial infertility (P<.001). No difference was noted in pregnancy rates according to fibroid characteristics. CONCLUSION(S): Fertility and pregnancy after laparoscopic myomectomy depend primarily on patient age, duration of infertility before myomectomy, and existence of associated infertility factors. PMID- 11476789 TI - Cellular events during fertilization failure in humans(2)(2). PMID- 11476790 TI - A polymorphism of the interleukin-1beta gene and idiopathic recurrent miscarriage. AB - OBJECTIVE: Proinflammatory cytokines have been described to be involved in the pathogenesis of idiopathic recurrent miscarriage (IRM). We investigated the association between IRM and a polymorphism in exon 5 of the interleukin-1beta gene (IL1B) and interleukin-1beta (IL-1beta) serum levels. DESIGN: Case control study. SETTING: Academic research institution. SUBJECTS: One hundred thirty-one women with a history of three or more consecutive pregnancy losses before 20 weeks' gestation and 68 healthy controls with at least two live births and no history of pregnancy loss. INTERVENTIONS: Peripheral venous puncture. MAIN OUTCOME MEASURES: An IL1B exon 5 (position +3953) gene polymorphism was analyzed by PCR amplification followed by restriction fragment length polymorphism analysis. IL-1beta serum levels were analyzed by a commercially available ELISA. RESULTS: Allele frequencies in women with IRM and controls were 77.9% and 80.8%, respectively, for the E1 allele (wild type), and 22.1% and 19.2%, respectively, for the E2 allele (mutant). No association between the E2 allele and the occurrence of IRM was found (P=.57, odds ratio =.83). Genotype frequencies and IL 1beta serum levels were not significantly different between the study group and the control group. CONCLUSIONS: This is the first report on an IL1B polymorphism in IRM. Although known to alter IL-1beta expression, the investigated IL1B polymorphism is not associated with IRM and increased serum levels in a large Caucasian population. PMID- 11476791 TI - Timing of testicular sperm retrieval procedures and in vitro fertilization intracytoplasmic sperm injection outcome. AB - OBJECTIVE: To compare the outcome of IVF-intracytoplasmic sperm injection (ICSI) using testicular spermatozoa obtained on the day of ovum pick-up (OPU) or on the day before OPU. DESIGN: Retrospective study. SETTING: An IVF clinic in a university hospital. PATIENT(S): Forty-seven IVF-ICSI cycles using testicular spermatozoa in 28 couples with the male partner suffering from nonobstructive azoospermia. INTERVENTION(S): Sperm retrieval was performed either on the OPU day (23 cycles in 19 patients; group A) or on the day before OPU (24 cycles in 15 patients; group B). Testicular sperm aspiration (TESA) was performed and followed by testicular sperm extraction (TESE) if no spermatozoa could be found. MAIN OUTCOME MEASURE(S): The presence of motile spermatozoa at the time of ICSI and fertilization and clinical pregnancy rates. RESULT(S): A similar proportion of motile spermatozoa (60.9% vs. 62.5%), fertilization rate (61.7% vs. 58.9%), and clinical pregnancy rate per transfer (34.8% and 29.2%) were obtained for groups A and B, respectively. CONCLUSION(S): Testicular sperm retrieval can be performed on the day before OPU without compromising success. Considerable medical and practical advantages may be offered by further advancement of testicular sperm retrieval procedures to 48 hours before OPU. This approach should thus be further evaluated. PMID- 11476792 TI - Comparison of several one-step home urinary luteinizing hormone detection test kits to OvuQuick. AB - OBJECTIVE: To compare clinical accuracy and ease of use for several of the new rapid one-step home urinary LH detection kits compared with the preexisting OvuQuick brand LH detection kit (designated as the standard). DESIGN: Prospective cohort study. SETTING: University-based infertility clinic. PATIENT(S): All women undergoing intrauterine inseminations at the clinic, regardless of infertility diagnosis, were offered enrollment during a 28-month study period. INTERVENTION(S): Each participant was supplied three one-step test kits (OvuQuick One-Step, ClearPlan Easy, and SureStep) in addition to a multistep OvuQuick test kit and instructed to run the tests in parallel on the same urine sample and to record the results. Urine testing was performed every 12 hours, beginning 3 days before the anticipated onset of an LH surge, and continued with the one-step kits every 12 hours for 48 hours after the surge was first detected by OvuQuick. Subjects also completed questionnaires evaluating the use of each test kit. MAIN OUTCOME MEASURE(S): Correlation of LH surge detection by one-step kits in comparison to surge detection by OvuQuick. Satisfaction and ease of use questionnaires. RESULT(S): Sixty-three volunteers returned useable data, from which 81 evaluable cycles were analyzed. The majority of patients found the newer one-step kits to be easier to use and less time consuming than OvuQuick. The three one-step kits detected the LH surge within +/- one testing period (+/-12 hours) of detection by OvuQuick 68%-84% of the time. OvuQuick One-Step, with modified instructions allowing for an equal color intensity in the test and reference areas, had the highest correlation with OvuQuick (84%). However, with a study power (alpha = 0.05, beta = 0.10) sufficient to detect a 10% difference between Ovuquick and each one-step kit, all one-step kits were statistically equivalent to each other. There was no cycle in which a one-step kit detected a positive LH surge but OvuQuick did not. CONCLUSION(S): One-step urinary LH kits are easier for patients to use than a multistep home urinary LH kit and have reasonable correlation with the multistep kit when used clinically for timing artificial inseminations. PMID- 11476793 TI - Successful in vitro fertilization-surrogate pregnancy in a patient with ovarian transposition who had undergone chemotherapy and pelvic irradiation. AB - OBJECTIVE: To present a case of IVF-surrogate pregnancy in a patient with ovarian transposition who had undergone chemotherapy and total pelvic irradiation. DESIGN: Case report. SETTING: Teaching hospital. PATIENT(S): A 29-year-old woman who had undergone Wertheim's hysterectomy for a bulky carcinoma of the uterine cervix. INTERVENTION(S): Ovarian transposition before chemotherapy and total pelvic irradiation. Standard IVF treatment, transabdominal oocyte retrieval, and transfer to the surrogate mother. MAIN OUTCOME MEASURE(S): Results of the IVF cycle. RESULT(S): A twin pregnancy at the first cycle and two live newborns. CONCLUSION(S): This is the first reported case of ovulation induction and oocyte retrieval performed on a transposed ovary. PMID- 11476794 TI - Resumption of luteinizing hormone pulsatility and hypogonadotropic hypogonadism after endoscopic ventriculocisternostomy in a hydrocephalic patient. AB - OBJECTIVE: To study gonadotropin pulsatility before and after surgical cure of hydrocephalus. DESIGN: Case report. SETTING: Department of Endocrinology and Centre d'Investigations Cliniques, Necker Hospital, Paris, France. PATIENT(S): A 29-year-old woman who presented with secondary amenorrhea. INTERVENTION(S): The patient underwent an endoscopic ventriculocisternostomy that led to restoration of normal menses and resolution of hypogonadism. MAIN OUTCOME MEASURE(S): A gonadotropin pulse study was performed before and 2 and 5 months after surgery. RESULT(S): No LH pulse was observed before surgery. Emergence of pulsatility was observed 2 months after surgery, and pulses became clearly individualized after 5 months. CONCLUSION(S): This observation strongly suggests that amenorrhea, in case of chronic hydrocephalus, is indeed due to a hypothalamic dysfunction of the GnRH pulse generator. PMID- 11476795 TI - A prospective randomized study comparing intramuscular progesterone and 17alpha hydroxyprogesterone caproate in patients undergoing in vitro fertilization-embryo transfer cycles. AB - OBJECTIVE: To compare the effectiveness of i.m. P and 17alpha-hydroxyprogesterone caproate (17-HPC) for luteal phase support, in patients undergoing IVF-ET cycles. DESIGN: Prospective, randomized study. SETTING: Patients undergoing IVF-ET in our Centers. PATIENT(S): The inclusion criteria were the use of GnRH down-regulation and aged <40 years. INTERVENTION(S): A total of 300 cycles were randomly treated with either 17-HPC (341 mg every 3 days) or P (50 mg daily). MAIN OUTCOME MEASURE(S): The outcomes of IVF in both study groups were evaluated for biochemical pregnancy, miscarriage, clinical pregnancy, and ongoing pregnancy. RESULT(S): No difference was found in the main outcome parameters considered. CONCLUSION(S): Although the results of the study encourage the use of 17-HPC for luteal phase support in patients undergoing IVF-ET program, more studies are necessary to support the hypothesis that it can replace i.m. P-in-oil. PMID- 11476796 TI - Periovulatory serum human chorionic gonadotropin (hCG) concentrations following subcutaneous and intramuscular nonrecombinant hCG use during ovulation induction: a prospective, randomized trial. AB - OBJECTIVE: To describe serum levels of human chorionic gonadotropin (hCG) as a function of hCG injection method (subcutaneous vs. intramuscular) among infertile women undergoing ovulation induction. DESIGN: Prospective, randomized clinical trial. SETTING: Major urban infertility referral center. PATIENT(S): Women presenting for infertility evaluation and ovulation induction. INTERVENTION(S): Controlled ovarian hyperstimulation was followed by 5,000 IU urinary (nonrecombinant) hCG injection, given intramuscularly (i.m.) or subcutaneously (s.c.). MAIN OUTCOME MEASURE(S): Serum hCG levels measured 24 hours after administration of hCG, and patient tolerability of injected hCG. RESULT(S): There were no statistically significant differences in age or body mass index (BMI) among patients receiving hCG s.c. (n = 13) or i.m. (n = 15). Mean [IQR (25; 75)] serum hCG levels in the s.c. and i.m. groups were 171.7 [27.0; 207.0] and 142.2 [102.5; 157.5] mIU/mL, respectively. No adverse events were registered by any patient receiving hCG by either injection method. In this non-IVF population, two pregnancies were established in each subgroup (4 of 28, or approximately 14% pregnancy rate). CONCLUSION(S): The s.c. administration of 5,000 IU hCG (reconstituted in vol. = 0.5 mL) was well tolerated by all women in this study and was associated with postinjection serum hCG levels similar to those observed after administration of an equivalent i.m. hCG dose. This investigation suggests that clinical use of s.c. hCG is suitable for lean women (e.g., BMI <30) undergoing ovulation induction, but additional data are needed to study the appropriateness of s.c. hCG administration in heavier patients. PMID- 11476797 TI - Successful pregnancies and deliveries after a simple vitrification protocol for day 3 human embryos. AB - OBJECTIVE: To evaluate the efficacy and efficiency of freezing cleaved human embryos through vitrification. DESIGN: Clinical study of vitrification of human embryos. SETTING: Assisted reproductive technology centers. PATIENT(S): Thirty six patients undergoing IVF-ICSI treatment whose surplus embryos were frozen. INTERVENTION(S): Two hundred fifteen surplus embryos vitrified, subsequently thawed, and transferred in natural or controlled cycles. MAIN OUTCOME MEASURE(S): Embryo survival rate after thawing and resultant patient pregnancy rate. RESULT(S): From the 215 vitrified and thawed embryos, 106 survived, with an overall embryo survival rate of 49.3%. The survival rate was higher when embryos were vitrified at the eight-cell stage compared with at the six to seven-cell and six-cell stages (79.2%, 39.7%, and 21.1%, respectively). On average, 2.9 +/- 1.2 embryos per patient were transferred, resulting in 11 pregnancies (30.5%), with an implantation rate of 10.4% per embryo transferred. CONCLUSION(S): Ultrarapid embryo freezing by vitrification of eight-cell stage embryos is a reliable method, as evidenced by high rates of embryo survival and pregnancy, making it a superior alternative to the conventional slow-cooling method. PMID- 11476798 TI - Reduction of ovarian torsion 1 week after embryo transfer in a patient with bilateral hyperstimulated ovaries. AB - OBJECTIVE: To report a rare case of unilateral ovarian torsion 1 week after embryo transfer in a patient with bilateral hyperstimulated ovaries. DESIGN: Case report and literature review. SETTING: Reproductive Endocrine division in a university teaching hospital. PATIENT(S): Infertility patients undergoing IVF-ET. INTERVENTION(S): Laparoscopic reduction of adnexa 1 week after ET. MAIN OUTCOME MEASURE(S): Successful preservation of the affected adnexa. RESULT(S): Delivery of 3.324 kg male infant with preservation of the affected ovary. CONCLUSION(S): Untwisting of the affected ovary at laparoscopy without aspiration reduction of cystic masses is appropriate. The outcome of the pregnancy (even very early) in patients with torsion of the adnexa may be favorable after a laparoscopic unwinding of the affected adnexa. PMID- 11476799 TI - Awareness of infertility in men with cystic fibrosis. PMID- 11476800 TI - Efficacy of clomiphene citrate and cyclofenil for infertile women with normal ovulatory cycles. PMID- 11476801 TI - Androgen receptor trinucleotide polymorphism in endometriosis. PMID- 11476802 TI - Are there indications for ovarian tissue cryopreservation? PMID- 11476803 TI - Prevalence of human herpesvirus types 1-8 in the semen of infertility patients and correlation with semen parameters. PMID- 11476804 TI - Intracytoplasmic sperm injection with polyvinylpyrrolidone: a potential risk. PMID- 11476805 TI - Colonic injury: does colostomy still have a place? PMID- 11476806 TI - Evolving concepts in the management of colonic injury. AB - PURPOSE: The management of colonic injury has changed in recent years. This study sought to evaluate current surgical management of injuries to the colon in a busy urban trauma centre, in the light of our increasing confidence in primary repair and evolving understanding of the concepts and practice of damage control surgery. METHODS: A retrospective analysis was made of consecutive patients presenting with colonic injury from January 1 to December 31 1998. Patients without full-thickness lesions of the colon were excluded, as were patients who died within 24 h of admission. Demographic data, wounding patterns and clinical course were studied. RESULTS: One hundred twenty-seven patients were analyzed. Management without colostomy was achieved in 84% of cases. Patients who underwent diversion of the faecal stream had increased morbidity and hospital stay compared to equivalent patients who were repaired primarily. The important subgroup of patients who underwent damage control or abbreviated laparotomy is discussed. CONCLUSION: This study further strengthens the validity of direct repair or resection and primary anastomosis for colonic injury. Strategies to deal with the subgroup of patients at very high risk of postoperative complications are suggested. PMID- 11476807 TI - Diaphragmatic rupture and the association with occupant position in right-hand drive vehicles. AB - Ruptured diaphragm following blunt trauma occurs with an incidence of 3 to 8% with right-sided rupture recognised with increasing frequency. This study aimed to investigate the influence of occupant position in right-hand drive (RHD) vehicles on the side of diaphragmatic injury. A retrospective analysis of the Scottish Trauma Audit Group database was performed to gather data on blunt diaphragmatic lacerations. Police records were also searched to ascertain the point of impact in the accidents studied. In total, 35 patients were studied, 25 drivers and 10 front-seat passengers. The incidence of right-sided rupture was 40% in drivers and 20% in FSPs. The incidence of associated pulmonary contusion, rib fracture and liver injury was also higher in drivers. Given the small sample size, these differences were not statistically significant, but they show an interesting trend. The right side of a driver's body is more exposed to injury in RHD vehicles, a fact that explains the significant association between driver's side impact and right-sided rupture. As right-sided injury is more difficult to detect, it is important that a high index of suspicion is maintained, especially when managing drivers from RHD vehicles. PMID- 11476808 TI - The anatomy of a shark attack: a case report and review of the literature. AB - Shark attacks are rare but are associated with a high morbidity and significant mortality. We report the case of a patient's survival from a shark attack and their subsequent emergency medical and surgical management. Using data from the International Shark Attack File, we review the worldwide distribution and incidence of shark attack. A review of the world literature examines the features which make shark attacks unique pathological processes. We offer suggestions for strategies of management of shark attack, and techniques for avoiding adverse outcomes in human encounters with these endangered creatures. PMID- 11476809 TI - Efficacy of prehospital critical care teams for severe blunt head injury in the Australian setting. AB - OBJECTIVE: To determine whether prehospital critical care teams (CCT) would result in improved functional outcomes for road trauma related severe head injury in the Australian setting, when compared with standard advanced life support measures provided by paramedics. METHODS: Retrospective review of 250 patients treated by paramedics and 46 patients treated by CCT transported directly from the accident scene, with a prehospital Glasgow coma scale (GCS)< or =8. RESULTS: CCT-treated patients had longer median prehospital times (113 versus 45 min, P<0.001), and a higher prehospital intubation rate (100% versus 36%, P<0.001) than paramedic-treated patients. On multivariate analysis, revised trauma score > or =4.45 (odds ratio [OR] 2.31, 95% CI: 1.15-4.65), lower injury severity score (OR 1.04, 95% CI: 1.02-1.06), age< or =25 years (OR 1.76, 95% CI: 1.13-2.75), absence of an acute subdural haematoma (OR 3.36, 95% CI: 1.89-5.95) and prehospital treatment by a CCT (OR 2.70, 95% CI: 1.48-4.95) independently predicted better outcome. CONCLUSION: The range of advanced interventions provided by the CCT were associated with improved functional outcome. Further studies are required to determine the individual factors responsible. PMID- 11476810 TI - Current cementing techniques in hip hemi-arthroplasty. AB - To assess the use of modern cementing techniques in hip hemi-arthroplasty a postal questionnaire was sent during 1998 to all British Orthopaedic Training Association (B.O.T.A.) members regarding use of cement, type of cement, bone preparation and cementing technique. The results of this survey were compared to a similar survey in 1994 to assess any change in practice. Two hundred and eighty six trainees responded to the 1998 survey, 352 to the 1994 survey. The use of uncemented prostheses had decreased from 31.3 in 1994 to 21.7% in 1998. Normal viscosity Palacos cement remains the most common cement in use, 64.3% in 1998. The use of antibiotic loaded cement has increased from 53.7 in 1994 to 67.9% in 1998. For bone preparation 47.3% of trainees in 1998 used a modern technique (syringe irrigation/pulsed lavage, brushing, gauze packing) compared to 35.1% in 1994. Modern cement insertion (retrograde gun, cement restriction and sustained pressure) was carried out by 39.3% in 1998 compared to 28.5% in 1994. Overall 27.2% of trainees used modern cementing techniques in hip hemi-arthroplasty, compared to 19.4% in 1994. Modern cementing techniques are used by a minority of British orthopaedic trainees, but in comparison to 1994 their use has increased. PMID- 11476811 TI - Re-operations following Austin Moore hemiarthroplasty: a district hospital experience. AB - The Austin Moore prosthesis is commonly used for displaced femoral neck fractures. In this study the rate of re-operation following Austin Moore hemiarthroplasty was studied. Ten out of 88 cases (11.36%) were re-operated on within the first year following the original operations. The indications for re operation were infection (4.5%), dislocation (3.4%) and loose prosthesis (3.4%). The rate of infection found in this study was higher than other published series and could be attributed to the lack of laminar flow ventilation system and the use of cotton drapes and surgical gowns while performing these operations. We recommend that Austin Moore hemiarthroplasty should be carried out in theatres equipped with a laminar flow ventilation system and to use waterproof gowns and drapes to limit the number of deep infections and therefore lower the re operation rate. PMID- 11476812 TI - Timing of operative intervention in the management of acutely fractured ankles and the cost implications. AB - A study of the length of the time between the diagnosis of an ankle fracture and operative intervention and the length of subsequent hospital stay was undertaken. The delay in operative fixation beyond 24 h from injury was associated with a lengthening of stay. The cost implication of a longer stay was assessed. Eighty seven patients with 87 fractures fulfilled the inclusion criteria of having an acute closed fracture of the ankle requiring open reduction and internal fixation (ORIF). There were 34 unimalleolar, 35 bimalleolar and 18 trimalleolar fractures. Only 47 (54%) of the patients were operated on within 24 h of injury, even though 74 had presented by 6 h and a further five by 24 h. The mean inpatient stay was 9.6 days for this early operation group. The patients who had their operation delayed were in hospital for a mean of 14 days, a significant difference (P<0.0001) (using Wilcoxon's Signed Rank test). The cost per patient per day of an acute trauma bed is estimated at pound sterlings 225. This translates into an average cost of pound sterlings 990 more per patient whose operation is delayed. We recommend that policies be put in place to provide early operative intervention for patients with fractured ankles as this would result in significant financial savings. PMID- 11476813 TI - Comparison of search strategies and quality of medical information of the Internet: a study relating to ankle sprain. AB - OBJECTIVES: to evaluate the quality of web sites on ankle sprain diagnosis and treatment and to assess the impact of Internet search expertise on quality of retrieved information. METHOD: two internet search strategies were conducted - one developed by an experienced information officer (expert's search) and the other based on the search terms used by orthopaedic medical staff (doctors' search). RESULTS: the expert's search revealed 32 web sites, of which nine were relevant, whereas the doctors' search revealed 61 web sites of which 27 were relevant. Of the relevant web sites in the expert's search, one complied with all quality criteria, whereas none of the web sites in the doctors' search complied with all criteria (11 vs. 0%, P=0.25). The web sites identified by expert's search had higher credibility (median scores 70 vs. 44, P=0.01) and accuracy of content (median scores 50 vs. 35, P=0.24). CONCLUSION: the quality of medical information on the internet is generally poor and information experts can capture higher quality web sites compared with doctors. PMID- 11476814 TI - The role of a fibular nail in the management of Weber type B ankle fractures in elderly patients with osteoporotic bone--a preliminary report. AB - Fibular nailing was performed in 11 Weber type B ankle fractures in elderly patients (average 67.2 years) with osteoporotic bones between March 1996 and February 2000. All the fractures were displaced with significant talar shift. Nine patients remained under follow-up (average 25.9 months). Results were good and excellent in 88% of cases by the modified Olerud and Molander scoring system. One patient developed post-operative ankle subluxation and required ankle fusion. We feel that fibular nailing is an important tool in managing specific types of ankle fracture. PMID- 11476815 TI - Peroneal tenosynovitis following ankle sprains. AB - Peroneal tenosynovitis has a variety of causes which include inversion injury of the ankle. It is rarely reported and frequently misdiagnosed. We report the operative findings and outcome of surgical decompression in 19 patients who clinically presented following inversion injury with features suggestive of peroneal tenosynovitis. A review of the literature of this pathology is presented. PMID- 11476816 TI - The operative management of displaced intra-articular fractures of the calcaneum: a two-centre study using a defined protocol. AB - The management of intra-articular calcaneal fractures remains controversial with strong arguments supporting both conservative and operative management. This study assesses the results of open reduction and internal fixation (ORIF) of displaced intra-articular fractures of the calcaneum in two independent centres where the indications for operative management had been strictly defined.Forty seven patients (51 fractures) who had sustained such injuries underwent an ORIF performed by one of the senior surgeons using the technique described by Eastwood et al. [JBJS 75-B(1993)189] All of the fractures were assessed by plain X-ray and CT scan and graded according to the Sanders classification [Clin. Orthop. 290(1993)97]. Patients were assessed at a minimum of 2 years post-surgery both clinically and with a standardised questionnaire based on that described by Kerr et al. [Injury 27(1996)35]. The mean age at operation was 42 and 50 years for the two centres, and the mean follow-up was 44 months. Seventy-six per cent of the patients were male. Eighty-eight per cent of the injuries were due to a fall from a height. Fifteen patients had contralateral foot/ankle injuries. Thirty-four of 46 patients were in employment at the time of the injury, 24 of these were in physical jobs and 20 sustained their injury whilst at work.Overall, the satisfaction rate was 90%. Ninety-four per cent of patients in work returned to work at a median of 6 months. Bilateral injuries were associated with a significantly poorer outcome as were those with heel pad pain. Delay to operation greater than 14 days was associated with a higher infection rate. Smoking was not related to infection rate. PMID- 11476817 TI - Wooden splinter in the foramen magnum. PMID- 11476818 TI - Contralateral acute subdural haematoma after burr-hole for chronic subdural haematoma. PMID- 11476819 TI - Odontoid fractures complicated by fractures of the posterior arch of the atlas in the elderly over 85 years with severe thoracic kyphosis secondary to osteoporosis. PMID- 11476820 TI - Cauda equina syndrome caused by delayed traumatic spinal subdural haematoma. PMID- 11476821 TI - A case of congenital eventration of the diaphragm mimicking traumatic diaphragmatic rupture. PMID- 11476822 TI - Traumatic diverticulosis and seromuscular rupture of the sigmoid colon as a result of non-penetrating abdominal trauma. PMID- 11476823 TI - Non-operative treatment of duodenal perforation secondary to blunt abdominal trauma. PMID- 11476824 TI - The Volkmann's spoon technique for removal of the elusive foreign body. PMID- 11476825 TI - Stem-cell research: drawing the line. PMID- 11476826 TI - Prions and proteins: distinguishing between conformations. PMID- 11476827 TI - Social capital and health: tip-toeing through the minefield of evidence. PMID- 11476828 TI - Biosafety consequences of eradication of wild-type polioviruses. PMID- 11476829 TI - Mechanism of complex regional pain syndrome: no longer excessive sympathetic outflow? PMID- 11476831 TI - Treatment of seborrhoeic dermatitis with oral terbinafine? PMID- 11476832 TI - Tissue distribution of protease resistant prion protein in variant Creutzfeldt Jakob disease using a highly sensitive immunoblotting assay. AB - BACKGROUND: Variant Creutzfeldt-Jakob disease (vCJD) has a pathogenesis distinct from other forms of human prion disease: disease-related prion protein (PrP(Sc)) is readily detectable in lymphoreticular tissues. Quantitation of risk of secondary transmission, and targeting of risk reduction strategies, is limited by lack of knowledge about relative prion titres in these and other peripheral tissues, the unknown prevalence of preclinical vCJD, and a transmission barrier which limits the sensitivity of bioassay. We aimed to improve immunoblotting methods for high sensitivity detection of PrP(Sc) to investigate the distribution of PrP(Sc) in a range of vCJD tissues. METHODS: We obtained tissues at necropsy from four patients with neuropathologically confirmed vCJD and from individuals without neurological disease. Tissues were analysed by sodium phosphotungstic acid precipitation of PrP(Sc) and western blotting using high sensitivity enhanced chemiluminescence. FINDINGS: We could reliably detect PrP(Sc) in the equivalent of 50 nL 10% vCJD brain homogenate, with a maximum limit of detection equivalent to 5 nl. PrP(Sc) could be detected in tissue homogenates when present at concentrations 10(4)-10(5) fold lower than those reported in brain. Tonsil, spleen, and lymph node were uniformly positive for PrP(Sc) at concentrations in the range of 0.1-15% of those found in brain: the highest concentrations were consistently seen in tonsil. PrP(Sc) was readily detected in the retina and proximal optic nerve of vCJD eye at levels of 2.5 and 25%, respectively of those found in brain. Other peripheral tissues studied were negative for PrP(Sc) with the exception of low concentrations in rectum, adrenal gland, and thymus from a single patient with vCJD. vCJD appendix and blood (Buffy coat fraction) were negative for PrP(Sc) at this level of assay sensitivity. INTERPRETATION: We have developed a highly sensitive immunoblot method for detection of PrP(Sc) in vCJD tissues that can be used to provide an upper limit on PrP(Sc) concentrations in peripheral tissues, including blood, to inform risk assessment models. Rectal and other gastrointestinal tissues should be further investigated to assess risk of iatrogenic transmission via biopsy instruments. Ophthalmic surgical instruments used in procedures involving optic nerve and the posterior segment of the eye, in particular the retina, might represent a potential risk for iatrogenic transmission of vCJD. Tonsil is the tissue of choice for diagnostic biopsy and for population screening of surgical tissues to assess prevalence of preclinical vCJD infection within the UK and other populations. PMID- 11476833 TI - Comparison of UK paediatric cardiac surgical performance by analysis of routinely collected data 1984-96: was Bristol an outlier? AB - BACKGROUND: Reports of high mortality after paediatric cardiac surgery at the Bristol Royal Infirmary, UK, led to the establishment of an independent public inquiry. A key question was whether or not the mortality statistics in Bristol were unusual compared with other specialist centres. To answer this question, we did a retrospective analysis of mortality in the UK using two datasets. METHODS: Data from the UK Cardiac Surgical Register (CSR; January, 1984, to March, 1996) and Hospital Episode Statistics (HES; April, 1991, to December, 1995) were obtained for all 12 major centres in which paediatric cardiac surgery is done in the UK. The main outcome measure was mortality within 30 days of a cardiac surgical procedure. We estimated excess deaths in Bristol using a random-effects model derived from the remaining 11 centres. Additionally, a sensitivity analysis was done and case-mix examined. FINDINGS: For children younger than 1 year, in open operations, the mortality rate in Bristol was around double that of the other centres during 1991-95: within the CSR, there were 19.0 excess deaths (95% interval 2-32) among 43 deaths; and in HES, there were 24.1 excess deaths (12-34) among 41 deaths recorded. There was no strong evidence for excess mortality in Bristol for closed operations or for open operations in children older than 1 year. INTERPRETATION: Our results suggest that Bristol was an outlier, and we do not believe that statistical variation, systematic bias in data collection, case mix, or data quality can explain a divergence in performance of this size. PMID- 11476835 TI - Effect of environmental manipulation in pregnancy and early life on respiratory symptoms and atopy during first year of life: a randomised trial. AB - BACKGROUND: Asthma places huge demands on health-care services, and its prevalence is increasing. Reduction of exposure to environmental allergens could offer a realistic chance for primary prevention. Our aim was to ascertain whether or not living in a low-allergen environment reduces the risk of asthma and atopic diseases in infants. METHODS: We assigned infants to four risk groups according to parental atopic status. We enrolled 291 high-risk couples (both parents atopic, no pets) into a prospective, prenatally randomised, cohort study, and allocated them to environmental manipulation, in which measures to reduce prenatal and postnatal allergen exposure were undertaken (active HRA) (n=145) or no intervention (control HRC) (n=146). Two further prospective groups were studied: 161 high-risk infants with pets in the home (HRP group) and 168 low-risk infants, whose parents were both non-atopic (LR group). The main outcome measures were signs and symptoms of atopic disease at 1 year of age. FINDINGS: 103 families dropped out or were lost to follow up. At age 1 year we followed-up 133 HRA, 118 HRC, 140 HRP, and 126 LR infants. Children in the HRA group were less likely to have respiratory symptoms during the first year of life than those in the HRC group. The most pronounced differences were in the relative risks for severe wheeze with shortness of breath (relative risk 0.44 [95% CI 0.20-1.00]), prescribed medication for the treatment of wheezy attacks (0.58 [0.36-0.95]), and wheezing after vigorous playing, crying, or exertion (0.18 [0.04-0.79]). Probability of respiratory symptoms in HRC and HRP infants was similar, whereas it was much lower in the LR than in the HRC group. Cat ownership was significantly associated with sensitisation to cats (24.6 [3.04-199.05]; p=0.003). INTERPRETATION: Environmental manipulation reduces some respiratory symptoms in the first year of life in high-risk infants. Further follow up is needed, however, to ascertain whether living in a low-allergen environment reduces allergy and asthma in later life. PMID- 11476836 TI - Income inequality, the psychosocial environment, and health: comparisons of wealthy nations. AB - BACKGROUND: The theory that income inequality and characteristics of the psychosocial environment (indexed by such things as social capital and sense of control over life's circumstances) are key determinants of health and could account for health differences between countries has become influential in health inequalities research and for population health policy. METHODS: We examined cross-sectional associations between income inequality and low birthweight, life expectancy, self-rated health, and age-specific and cause-specific mortality among countries providing data in wave III (around 1989-92) of the Luxembourg Income Study. We also used data from the 1990-91 wave of the World Values Survey (WVS). We obtained life expectancy, mortality, and low birthweight data from the WHO Statistical Information System. FINDINGS: Among the countries studied, higher income inequality was strongly associated with greater infant mortality (r=0.69, p=0.004 for women; r=0.74, p=0.002 for men). Associations between income inequality and mortality declined with age at death, and then reversed among those aged 65 years and older. Income inequality was inconsistently associated with specific causes of death and was not associated with coronary heart disease (CHD), breast or prostate cancer, cirrhosis, or diabetes mortality. Countries that had greater trade union membership and political representation by women had better child mortality profiles. Differences between countries in levels of social capital showed generally weak and somewhat inconsistent associations with cause-specific and age-specific mortality. INTERPRETATION: Income inequality and characteristics of the psychosocial environment like trust, control, and organisational membership do not seem to be key factors in understanding health differences between these wealthy countries. The associations that do exist are largely limited to child health outcomes and cirrhosis. Explanations for between country differences in health will require an appreciation of the complex interactions of history, culture, politics, economics, and the status of women and ethnic minorities. PMID- 11476837 TI - Imaging of onset and progression of Alzheimer's disease with voxel-compression mapping of serial magnetic resonance images. AB - BACKGROUND: Early diagnosis and monitoring of the progression of Alzheimer's disease is important for the development of therapeutic strategies. To detect the earliest structural brain changes, individuals need to be studied before symptom onset. We used an imaging technique known as voxel-compression mapping to localise progressive atrophy in patients with preclinical Alzheimer's disease. METHODS: Four symptom-free individuals from families with early-onset Alzheimer's disease with known autosomal dominant mutations underwent serial magnetic resonance imaging (MRI) over 5-8 years. All four became symptomatic during follow up. 20 individuals with a clinical diagnosis of probable Alzheimer's disease and 20 control participants also underwent serial MR imaging. A non-linear fluid matching algorithm was applied to register repeat scans onto baseline imaging. Jacobian determinants were used to create the voxel-compression maps. FINDINGS: Progressive atrophy was revealed in presymptomatic individuals, with posterior cingulate and neocortical temporoparietal cortical losses, and medial temporal lobe atrophy. In patients with known Alzheimer's disease, atrophy was widespread apart from in the primary motor and sensory cortices and cerebellum, reflecting the clinical phenomenology. INTERPRETATION: Voxel-compression maps confirmed early involvement of the medial temporal lobes, but also showed posterior cingulate and temporoparietal cortical losses at presymptomatic stage. This technique could be applied diagnostically and used to monitor the effects of therapeutic intervention. PMID- 11476838 TI - The heavy smoking football fan who had an epileptic fit. PMID- 11476839 TI - Linezolid resistance in a clinical isolate of Staphylococcus aureus. AB - The new oxazolidinone antimicrobial, linezolid, has been approved for the treatment of infections caused by various gram-positive bacteria, including meticillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Although instances of linezolid resistance in VRE have been reported, resistance has not been encountered among clinical isolates of S aureus. We have characterised an MRSA isolate resistant to linezolid that was recovered from a patient treated with this agent for dialysis-associated peritonitis. PMID- 11476840 TI - Detection of variant Creutzfeldt-Jakob disease infectivity in extraneural tissues. AB - Abnormal accumulations of prion protein (PrP) can be detected in the spleen, lymph nodes, and tonsils of patients with variant Creutzfeldt-Jakob disease (vCJD). Therefore, it has been assumed, but not shown, that these tissues harbour infectivity, which in turn presents the potential for iatrogenic spread through surgery. Here, we show and measure levels of infectivity in spleen and tonsil from two patients with vCJD, by bioassay in intracerebrally inoculated RIII mice. Similar bioassays failed to detect infectivity in buffy coat and plasma. PMID- 11476841 TI - Association of germline mutation in the PTEN tumour suppressor gene and Proteus and Proteus-like syndromes. AB - The molecular aetiology of Proteus syndrome (PS) remains elusive. Germline mutations in PTEN cause Cowden syndrome and Bannayan-Riley-Ruvalcaba syndrome, which are hereditary hamartoma syndromes. Some features-eg, macrocephaly, lipomatosis, and vascular malformations-can be seen in all three syndromes. We examined PTEN in patients with PS and undefined Proteus-like syndromes (PS-like) and identified de-novo germline mutations in two of nine patients with PS and three of five patients with PS-like. Germline PTEN mutation analysis should be done in individuals with PS and PS-like because of its association with increased risk of cancer development and potential of germline-mutation transmission. PMID- 11476842 TI - Unexpected healing of cutaneous ulcers in a short child. AB - A boy aged 10 years was referred to the Paediatric Department of Milan University Hospital, Milan, Italy, with a long history of pain in the lower limbs, alleviated only by exposure to cold. His legs were swollen, with multiple cutaneous ulcers. He had severe painful crises, and was totally incapacitated. After the diagnosis of erythermalgia was made, numerous treatments were tried, but none were successful. After finding growth hormone (GH) deficiency, we started treatment with recombinant GH. He had immediate relief of pain and complete healing of ulcers. We postulate that the healing of the ulcers can be attributed to the GH-promoting effect on dermal connective tissue. PMID- 11476843 TI - Johns Hopkins takes responsibility for volunteer's death. PMID- 11476849 TI - Why doctors need to know about atoms. PMID- 11476850 TI - Presidential decision near on stem-cell research. PMID- 11476851 TI - French government tries to persuade more doctors to work in rural areas. PMID- 11476854 TI - WHO tackles hearing disabilities in developing world. PMID- 11476855 TI - Indian state launches decentralised health scheme. PMID- 11476856 TI - Legionnaire's outbreak in Spanish town may be largest ever. PMID- 11476857 TI - Medicinal marijuana users in Canada must carry identification cards. PMID- 11476858 TI - Type 1 diabetes: new perspectives on disease pathogenesis and treatment. AB - As our knowledge of type 1 (insulin-dependent) diabetes increases, so does our appreciation for the pathogenic complexity of this disease and the challenges associated with its treatment. Many new concepts about the pathogenesis of this disorder have arisen. The role of genetics versus environment in disease formation has been questioned, and the basis on which type 1 diabetes is characterised and diagnosed is the subject of much debate. Additionally, the care and treatment of patients with type 1 diabetes has seen a rapid evolution; with genetically engineered insulins, glucose monitoring devices, and algorithms all contributing to a decrease in disease-related complications. We focus this seminar on these changing views, and offer a new perspective on our understanding of the pathogenesis of type 1 diabetes and on principles for therapeutic management of patients with this disorder. PMID- 11476859 TI - Evidence based road safety: the Driving Standards Agency's schools programme. PMID- 11476860 TI - Fear and frustration--the Liverpool cholera riots of 1832. AB - Public mistrust in the medical profession is not new. We describe a series of street riots that took place in the city of Liverpool in north-west England in 1832 during a cholera epidemic. The disturbances were directed primarily against the local medical fraternity. The episode is of interest, since the same city recently experienced a similar crisis of confidence between doctors and public. On this occasion the cause was not cholera, but rather the reports from Alder Hey Children's Hospital that organ parts from deceased infants undergoing necropsy had been kept for several years without parental consent. PMID- 11476867 TI - GDNF recruits the signaling crew into lipid rafts. AB - Glial cell line-derived neurotrophic factor (GDNF) family ligands (GFL) are potent survival factors and regulators for central and peripheral neurons. GFLs bind to specific glycosyl phosphatidylinositol (GPI)-anchored co-receptors (GFRalpha1-alpha4), but signal through a common c-Ret receptor. Both GPI-anchored and soluble GFRalpha1 recruit c-Ret to lipid rafts following GDNF stimulation, where c-Ret interacts with different proteins than outside the rafts. Soluble GFRalpha1 mobilizes c-Ret to rafts by a different mechanism compared with GPI anchored GFRalpha1. PMID- 11476868 TI - Neurosteroids and brain sexual differentiation. AB - There is new evidence that the brain of developing songbirds can synthesize estradiol de novo. In males, this neurally derived estrogen might masculinize a connection within the neural song system. These results challenge traditional concepts about mechanisms of brain sexual differentiation and reveal a significant function for neurosteroids. PMID- 11476870 TI - Glial cells aid axonal target selection. AB - A key problem in developmental neurobiology is how axons home in on their correct target tissue and establish the correct synaptic contacts. Recent work shows that in the developing Drosophila visual system a population of distinct lamina glial cells ensures correct target layer selection of retinal axons. In the absence of lamina neurons, photoreceptor axons terminate their growth in the correct zone, but when glial cell migration into the lamina is disrupted, as in nonstop mutants, growth cones advance into deeper layers of the brain. PMID- 11476871 TI - A multi-talented secreted protein. PMID- 11476872 TI - Engineering a sensory circuit in the brain: multiple roles for a single gene. PMID- 11476883 TI - Rocking and rolling with Ca2+ channels. AB - Voltage-sensitive Ca2+ channels are a large family of related heterooligomers that couple cell excitability to intracellular signalling. Recent studies on mice carrying Ca2+ channel mutations, or in which Ca2+ channel subunits have been deleted, have provided important information about the roles carried out by these molecules in cardiovascular function, pain, epilepsy, migraine and deafness, in addition to further defining how Ca2+ channels regulate the physiology of excitable cells. PMID- 11476884 TI - Glial activation: a driving force for pathological pain. AB - Pain is classically viewed as being mediated solely by neurons, as are other sensory phenomena. The discovery that spinal cord glia (microglia and astrocytes) amplify pain requires a change in this view. These glia express characteristics in common with immune cells in that they respond to viruses and bacteria, releasing proinflammatory cytokines, which create pathological pain. These spinal cord glia also become activated by certain sensory signals arriving from the periphery. Similar to spinal infection, these signals cause release of proinflammatory cytokines, thus creating pathological pain. Taken together, these findings suggest a new, dramatically different approach to pain control, as all clinical therapies are focused exclusively on altering neuronal, rather than glial, function. PMID- 11476885 TI - Synaptic reverberation underlying mnemonic persistent activity. AB - Stimulus-specific persistent neural activity is the neural process underlying active (working) memory. Since its discovery 30 years ago, mnemonic activity has been hypothesized to be sustained by synaptic reverberation in a recurrent circuit. Recently, experimental and modeling work has begun to test the reverberation hypothesis at the cellular level. Moreover, theory has been developed to describe memory storage of an analog stimulus (such as spatial location or eye position), in terms of continuous 'bump attractors' and 'line attractors'. This review summarizes new studies, and discusses insights and predictions from biophysically based models. The stability of a working memory network is recognized as a serious problem; stability can be achieved if reverberation is largely mediated by NMDA receptors at recurrent synapses. PMID- 11476886 TI - Studying rhythmogenesis of breathing: comparison of in vivo and in vitro models. AB - In all mammalian species, breathing is controlled by a neuronal network within the lower brainstem. A component known as the ventral respiratory group produces rhythmic activity, which is transmitted to spinal motoneurons to produce a periodic contraction of respiratory muscles. A dispute about the mechanisms of 'normal' respiratory rhythm generation arose from the differences between experimental preparations that have been used to dissect the process. It is, therefore, essential to compare the various experimental approaches and to discuss the differences between experimental data. We conclude that the various preparations all have great value, but that they define different operational conditions of the network, including maturation of neurons and synaptic processes. We have taken note of these in formulating a 'maturational network burster model' for rhythm generation that includes most features of the existing models of respiratory rhythm generation. PMID- 11476887 TI - Regulation of Na+ channel distribution in the nervous system. AB - An important aspect of Na+ channel regulation is their distribution on neuronal membranes within the nervous system. The complexity of this process is brought by the molecular diversity of Na+ channels and differential regulation of their distribution. In addition, Na+ channel localization is a highly dynamic process depending on the status of the cell in vitro, and (patho)physiological condition of the organism in vivo. Nonetheless, the pharmacological manipulation of Na+ channel distribution should be possible and will hopefully bring safer and more potent medicines in the future. PMID- 11476888 TI - Analysis of complex brain disorders with gene expression microarrays: schizophrenia as a disease of the synapse. AB - The level of cellular and molecular complexity of the nervous system creates unique problems for the neuroscientist in the design and implementation of functional genomic studies. Microarray technologies can be powerful, with limitations, when applied to the analysis of human brain disorders. Recently, using cDNA microarrays, altered gene expression patterns between subjects with schizophrenia and controls were shown. Functional data mining led to two novel discoveries: a consistent decrease in the group of transcripts encoding proteins that regulate presynaptic function; and the most changed gene, which has never been previously associated with schizophrenia, regulator of G-protein signaling 4. From these and other findings, a hypothesis has been formulated to suggest that schizophrenia is a disease of the synapse. In the context of a neurodevelopmental model, it is proposed that impaired mechanics of synaptic transmission in specific neural circuits during childhood and adolescence ultimately results in altered synapse formation or pruning, or both, which manifest in the clinical onset of the disease. PMID- 11476890 TI - Biochemical signals and biological responses elicited by the focal adhesion kinase. AB - The focal adhesion kinase, FAK, is an important component of an integrin dependent signaling pathway, which functions to transmit signals from the extracellular matrix into the cytoplasm. FAK is an essential gene product, since the fak-/- mouse exhibits embryonic lethality. A number of important biological processes, including cell motility and cell survival, are controlled by integrin dependent signals and FAK has been implicated in regulating these processes. This review will focus upon recent findings providing insight into the mechanisms by which FAK transmits biochemical signals and elicits biological effects. PMID- 11476891 TI - A novel photosensitizer, 2-butylamino-2-demethoxy-hypocrellin B (2-BA-2-DMHB) - its photodynamic effects on HeLa cells: efficacy and apoptosis. AB - A novel hypocrellin congener, 2-butylamino-2-demethoxy-hypocrellin B (2-BA-2 DMHB) was found to be an effective photosensitizer. Compared with its parent compound hypocrellin B (HB), its absorption bands extended toward longer wavelength and the extinction coefficients raised to some degree (lambda(max) (nm) (log(epsilon)): 463 (4.06), 583 (4.09), 621 (4.10) for 2-BA-2-DMHB and 466 (4.06), 548 (3.70), 580 (3.52) for HB). And it also had a much higher photopotentiation factor than HB (i.e., more than 250 versus 10 at a dose of 24 J cm(-2) of red light on HeLa cells). This might be correlated to the higher ability of superoxide anion generation and the higher cellular uptake of 2-BA-2 DMHB. Meanwhile, the examinations of Hoechst 33342-labeled nuclei, DNA fragmentation on agarose gel and flow cytometry showed that 2-BA-2-DMHB induces apoptosis in photosensitized HeLa cells more quickly than HB, which might be correlated to the higher cellular uptake of 2-BA-2-DMHB and the difference of their cellular localization. The study suggested that 2-BA-2-DMHB was a well suited candidate for a new generation of photodynamic therapy photosensitizer. PMID- 11476892 TI - Distinct patterns of protein binding to the MAT2A promoter in normal and leukemic T cells. AB - The metabolism of S-adenosylmethionine (AdoMet), a key molecule in regulating T cell differentiation and proliferation, is different in normal and leukemic T cells. To delineate the basis for these differences we studied the transcriptional regulation of human methionine adenosyltransferase II (MAT II), which catalyzes AdoMet synthesis in these cells. Recently, we identified an Sp1 site in the proximal promoter of the MAT2A gene, which encodes the alpha2 catalytic subunit of MAT II, that is essential for the in vitro and in vivo promoter activity in Jurkat leukemic T cells, and that involves binding of the nuclear factors Sp2 and Sp3, but not Sp1. Here, the in vitro and in vivo activity of the proximal MAT2A promoter in normal resting, PHA-stimulated, and leukemic human T cells was compared. Significantly different patterns of protein factor interaction in the proximal region of the MAT2A promoter were found. Normal resting and activated T cells produced complexes of significantly lower molecular weight than those formed in leukemic T cells. Supershift studies coupled with analysis of proteins bound to the proximal promoter suggest that low levels of expression of Sp2 and Sp3 in normal T cells may be responsible for the difference in the in vitro promoter activity between normal and leukemic cells. Mutation of the key Sp1 site equally reduced the in vivo promoter activity in normal and malignant T cells; by contrast, it had significantly different effects on protein DNA interactions in normal and leukemic T cells. Together, the data support the idea that differences in protein-DNA interactions may contribute to significant differences in MAT2A regulation in normal and malignant cells. PMID- 11476893 TI - Identification of xanthine dehydrogenase/xanthine oxidase as a rat Paneth cell zinc-binding protein. AB - Paneth cells are zinc-containing cells localized in small intestinal crypts, but their function has not been fully elucidated. Previously, we showed that an intravenous injection of diphenylthiocarbazone (dithizone), a zinc chelator, induced selective killing of Paneth cells, and purified a zinc-binding protein in Paneth cells. In the present study, we further characterized one of these proteins, named zinc-binding protein of Paneth cells (ZBPP)-1. Partial amino acid sequences of ZBPP-1 showed identity with rat xanthine dehydrogenase (XD)/xanthine oxidase (XO). Anti-rat XD antibody (Ab) recognized ZBPP-1, and conversely anti ZBPP-1 Ab recognized 85 kDa fragment of rat XD in Western blotting. Messenger RNA and protein levels of XD were consistent with our previous data on the fluctuation of Paneth cell population after dithizone injection. Thus, ZBPP-1 is an 85 kDa fragment of XD/XO in Paneth cells. XD/XO in Paneth cells may play important roles in intestinal function. PMID- 11476894 TI - Monocyte-macrophage differentiation in three dimensional collagen lattice. AB - Human peripheral blood mononuclear cells (PBMC) upon transendothelial migration interact with subendothelial matrix components and differentiate into macrophages. In order to study whether the shape of the cells as dictated by the extracellular matrix can influence monocyte-macrophage (mo-m(phi)) differentiation, human PBMC were maintained in vitro on a three dimensional collagen I (COL I) lattice and studied for various macrophage specific functions, viz. endocytosis of [(125)I]acetyl bovine serum albumin (BSA), expression of specific cell surface antigens and expression of matrix metalloproteinases (MMPs). The cells maintained in three dimensional COL gel exhibited a higher rate of endocytosis of [(125)I]acetyl BSA than those on COL-coated plastic. FACS analysis showed that the mean fluorescence intensity (MFI) corresponding to monocyte specific LPS receptor CD14 was significantly decreased while MFI corresponding to macrophage specific transferrin receptor CD71 was significantly increased in cells maintained in vitro on three dimensional COL gel compared to two dimensional COL substrata. Expression of macrophage specific MMPs (gelatinase A and gelatinase B) was significantly high in cells maintained on COL gel than on COL I-coated plastic. Appearance of 67 kDa gelatinase in the COL gel suggested that induction as well as activation of MMPs occur when cells are maintained in a three dimensional environment. These results indicate that monocytes undergo a rapid rate of differentiation when maintained in vitro on three dimensional COL I lattice suggesting that apart from the chemical nature of the matrix, the shape of the cells as provided by the matrix also influences mo-m(phi) differentiation. PMID- 11476895 TI - Expression pattern of the type 1 sigma receptor in the brain and identity of critical anionic amino acid residues in the ligand-binding domain of the receptor. AB - The type 1 sigma receptor (sigmaR1) has been shown to participate in a variety of functions in the central nervous system. To identify the specific regions of the brain that are involved in sigmaR1 function, we analyzed the expression pattern of the receptor mRNA in the mouse brain by in situ hybridization. SigmaR1 mRNA was detectable primarily in the cerebral cortex, hippocampus, and Purkinje cells of cerebellum. To identify the critical anionic amino acid residues in the ligand binding domain of sigmaR1, we employed two different approaches: chemical modification of anionic amino acid residues and site-directed mutagenesis. Chemical modification of anionic amino acids in sigmaR1 with 1-ethyl-3-(3 dimethylaminopropyl)carbodiimide reduced the ligand-binding activity markedly. Since it is known that a splice variant of this receptor which lacks exon 3 does not have the ability to bind sigma ligands, the ligand-binding domain with its critical anionic amino acid residues is likely to be present in or around the region coded by exon 3. Therefore, each of the anionic amino acids in this region was mutated individually and the influence of each mutation on ligand binding was assessed. These studies have identified two anionic amino acids, D126 and E172, that are obligatory for ligand binding. Even though the ligand-binding function was abolished by these two mutations, the expression of these mutants was normal at the protein level. These results show that sigmaR1 is expressed at high levels in specific areas of the brain that are involved in memory, emotion and motor functions. The results also provide important information on the chemical nature of the ligand-binding site of sigmaR1 that may be of use in the design of sigmaR1 specific ligands with potential for modulation of sigmaR1-related brain functions. PMID- 11476896 TI - Long-term hormonal regulation of the cAMP-specific phosphodiesterases in cultured FRTL-5 thyroid cells. AB - Thyrotropin (TSH) and pharmacological agents that elevate intracellular cAMP concentrations potentiate the mitogenic response of FRTL-5 thyroid cells to insulin-like growth factor-I (IGF-I). This study was undertaken to determine the role of cAMP phosphodiesterases (PDEs) in this TSH-dependent regulation. Incubation of FRTL-5 cells with TSH, forskolin, or dibutyryl cAMP gradually induced the PDE activity, and treatment for 24 h produced a marked increase in type 4 high affinity cAMP PDEs. Under basal conditions, transcripts corresponding to PDE4A, PDE4B, PDE4C, and PDE4D were present. Stimulation for 24 h by TSH, forskolin or dibutyryl cAMP induced an increase in mRNA levels of PDE4B, PDE4D, and PDE4C. To understand the role of this cAMP-dependent PDE regulation in the potentiation of the mitogenic response to IGF-I, thymidine incorporation into DNA in response to IGF-I and TSH was measured in the absence or presence of PDE inhibitors. Exposure of the cells to 3-isobutyl-1-methylxanthine (IBMX) or RO 20 1724 had opposing effects on thymidine incorporation into DNA, depending on the stimulus applied. When IGF-I was used alone, both IBMX and RO 20-1724 potentiated IGF-I-stimulated thymidine incorporation. However, when IGF-I and TSH at high concentrations were used in combination, these PDE inhibitors blocked thymidine incorporation into DNA. In addition, these inhibitors depressed the synergistic increase in cyclin D1 and cyclin D- or cyclin E-associated cyclin-dependent kinase (CDK) activity that is induced by TSH and IGF-I. Increased CDK activities have been shown to play a crucial role in progression through the G(1)/S phase of the cell cycle. These data demonstrate that TSH produces marked changes in the cAMP degradative pathway of FRTL-5 cells by regulating the expression of cAMP PDEs. The regulation of the intracellular cAMP levels by this mechanism may contribute to the TSH- and IGF-I-dependent control of the entry into the S phase of the cell cycle through changes in the cyclin/CDK system in FRTL-5 cells. PMID- 11476897 TI - AlphaIIb's cytoplasmic domain is not required for ligand-induced clustering of integrin alphaIIbbeta3. AB - The platelet integrin alphaIIbbeta3 exhibits bidirectional signaling, in that intracellular messengers enable adhesive macromolecules to bind to its ectodomain, while ligation promotes the association of cytoskeletal proteins with its cytoplasmic domains. In order to understand the linkage between these distant regions, we investigated the effects of receptor occupancy on the solution structure of both full-length recombinant alphaIIbbeta3 and alphaIIbDelta991beta3, an integrin truncation mutant which lacks one cytoplasmic domain. Lysates of (35)S-labeled human A549 cells expressing either full-length alphaIIbbeta3 or alphaIIbDelta991beta3 were examined by sucrose density gradient sedimentation followed by immunoprecipitation to determine the distributions of integrin protomers and oligomers. Recombinant alphaIIbbeta3 exhibited a weight average sedimentation coefficient, S(w)=11.3+/-1.4 S with 73% sedimenting as protomers/dimers (9.1+/-1.0 S) and 27% as oligomers (15.4+/-0.4 S). Truncation mutant alphaIIbDelta991beta3 exhibited a similar pattern with 65% sedimenting as protomers/dimers. Upon ligation with eptifibatide, both full-length alphaIIbbeta3 and alphaIIbDelta991beta3 sedimented mainly at >14 S, indicating 2-3-fold increased oligomerization. Thus we have demonstrated that alphaIIb's cytoplasmic region is not required for integrin clustering, a key event in outside-in signaling. PMID- 11476898 TI - Synthetic peptides that inhibit binding of the myelin basic protein 85-99 epitope to multiple sclerosis-associated HLA-DR2 molecules and MBP-specific T-cell responses. AB - Copolymer 1 (Cop 1, poly [Y, E, A, K]) is a random synthetic amino acid copolymer effective in the treatment of relapsing forms of multiple sclerosis (MS), a disease that is linked to HLA-DR2 (DRB1*1501). In the present study various peptides, synthesized according to the binding motifs for both the immunodominant epitope of myelin basic protein (MBP) 85-99, a candidate autoantigen in MS, and Cop 1, differentially inhibited binding of these antigens to disease-associated HLA-DR2 (DRB1*1501) molecules. In particular, two peptides with residue K at position P-1, as referred to MBP 85-99, inhibited effectively the binding of both biotinylated MBP 85-99 and Cop 1 to HLA-DR2 molecules as well as IL-2 production by two MBP-specific HLA-DR2-restricted T-cell clones. These findings suggest the possible utility of these compounds or their more stable derivatives in treatment of MS. PMID- 11476899 TI - Major histocompatibility class one molecule associates with glucose regulated protein (GRP) 78 on the cell surface. AB - Glucose regulated protein 78 (GRP78) is a member of the heat shock protein (hsp) 70 family. It is an endoplasmic reticulum (ER) chaperone, whose function is generally thought to be limited to the structural maturation of nascent glycoproteins. However, recent observations have shown that ER chaperones, such as GRP78, display peptide-binding activity. These peptide-binding activities along with the observation that heat shock proteins associated with peptides can elicit antigen-specific CTL responses suggest additional roles for these proteins. In this study we provide evidence that GRP78 is not only resident in the ER, but also exists on the cell surface. Furthermore, using biochemical and imaging studies we have found that GRP78 associates with MHC class I on the cell surface. Its presence on the cell surface is not dependent on MHC class I expression. In the absence of MHC class I its cell surface expression is upregulated. PMID- 11476900 TI - Effect of phorbol ester and calcium ionophore on human thymocytes. AB - Positive selection of immature thymocytes is a developmental process in which TCR ligation with low avidity induces generation of mature T cells. In mouse thymocytes, CD4(+)8(+) double-positive (DP) cells which were treated with a proper combination of calcium ionophore ionomycin and phorbol 12-myristate 13 acetate (PMA) have been reported to differentiate into CD4 single positive cells. However, in human thymocytes the effects of PMA and ionomycin have remained unclear. Here we report that DP cells that were treated with PMA and ionomycin up regulated bcl-2 and down-regulated CD1 expression. However, CD3 expression remained low. This treatment induced prolonged CD4 down-regulation in DP cells which was an effect also seen in mature peripheral blood T cells. PMA/ionomycin treated DP cells showed high cell proliferation and resistance to dexamethasone induced apoptosis. These results indicate that PKC activation and calcium elevation may be part of the biochemical signals that induce positive selection of human DP cells and the system described in this paper may be a useful model to study the signals involved in the selection of human thymocytes. PMID- 11476901 TI - NK cells recover early and mediate cytotoxicity via perforin/granzyme and Fas/FasL pathways in umbilical cord blood recipients. AB - Umbilical cord blood (UCB) is now widely accepted as a source of stem cells in patients with malignant hematologic and genetic disorders. We have recently reported that in a series of 30 pediatric UCB transplant recipients comparable outcome to that anticipated with other unrelated stem cell sources. In our series, however, the probability of GVHD for grade III-IV was 9% and no UCB recipient developed chronic GVHD. The reason for the low incidence of GVHD after UCB transplantation is not fully understood. Because functional NK cells are among the first population of lymphocytes to be detected in UCB transplant recipients, 2 months post-transplant on average, we wanted to establish whether NK cells could be implicated in reducing the risk of GVHD. Here, we confirm that early NK cells detected in UCB transplant recipients activate the granzyme/perforin lytic pathway and, in addition, they can mediate Fas/Fas ligand (FasL) activity, a finding not previously reported. Both pathways develop simultaneously and are detectable months before the other lymphocytes, notably CD8 are fully functional. Our contention, therefore, is that the low GVHD observed in UCB recipients may be partially due to early NK cells. PMID- 11476902 TI - Identification of p53 peptides recognized by CD8(+) T lymphocytes from patients with bladder cancer. AB - In many types of cancer, p53 frequently accumulates in tumor cells and anti-p53 antibodies can be detected. However, only four CD8(+) T-cell epitopes from p53 have been identified in humans so far. To further analyze the development of a T cell response against p53, peptides having binding motifs specific for HLA-A1, A2, -A3, -A24, -B7, -B35, -B44, and -B51 molecules have been defined. The HLA binding capacity of those peptides was tested, and the stability of formed complexes was defined. Thirteen peptides that bound to HLA-A24 and -B44 molecules are presented. The positive peptides were then used to detect the anti-p53 response of CD8(+) T lymphocytes from patients with bladder cancer. Six peptides, presented by HLA-A2, -B51, or -A24, were able to stimulate T cells from two patients (among 16) with tumor cells that strongly accumulated p53. On the contrary, p53 peptides systematically failed to stimulate T cells from healthy donors or patients with low or undetectable levels of p53 in their tumor cells. These results have led to the identification of four new potential T CD8(+) epitopes from p53: 194-203 associating with HLA-B51 and 204-212, 211-218, and 235 243 associating with HLA-A24. PMID- 11476903 TI - In vitro granuloma formation, NO production and cytokines profile from human mononuclear cells induced by fractionated antigens of Paracoccidioides brasiliensis. AB - Paracoccidioides brasiliensis causes a chronic granulomatous mycosis prevalent in South America, and cell-mediated immunity represents the main mode of protection against this fungal infection. We investigated in vitro the response of peripheral blood mononuclear cells (PBMC) from paracoccidioidomycosis (PCM) patients presenting different clinical forms to antigenic fractions from P. brasiliensis yeast cell lysate (PbAg). These fractions designated F0 to FV were obtained using anion-exchange chromatography on a FPLC system. Our studies showed variation in the cellular responses induced by different antigenic fractions. The fraction F0 caused significant decrease in cellular proliferation, granuloma formation, accompanied by significant elevation in the production of IL-10. The fractions FII and FIII increased in vitro granuloma formation associated with high production of TNF-alpha. Besides that, FII and FIII evoked decrease in NO production but not F0 that induced very high levels, among patients with PCM from acute form. The findings suggest that P. brasiliensis antigenic components participate in the modulation or activation of PBMC response in PCM, and IL-10 and NO could be important in the regulation of in vitro granuloma formation. PMID- 11476904 TI - HLA class I and II in Lambert-Eaton myasthenic syndrome without associated tumor. AB - Lambert-Eaton myasthenic syndrome (LEMS) is an autoimmune disorder, in which antibodies against voltage-gated calcium channels located at nerve terminals cause muscle weakness and autonomic dysfunction. In approximately half of the patients the autoimmune process is initiated by a tumor. In the other half of patients no tumor is found and the etiology is unknown. The aims of this study were to investigate the strength of HLA-associations with nontumor LEMS (NT-LEMS) and to study the relation of HLA-haplotypes with age at onset of LEMS and other clinical features. Therefore, typing of HLA class I and II was performed in 19 patients with NT-LEMS, who were clinically evaluated. NT-LEMS was significantly associated with alleles of both HLA-class I (i.e. HLA-B8) as well as -class II (i.e. HLA-DR3 and -DQ2). HLA-B8+ patients had significantly younger age at onset of LEMS and tended to be female. This study shows that HLA-class I haplotype is associated with a distinct phenotype in NT-LEMS. PMID- 11476905 TI - Class II allele and haplotype frequencies in Mexican systemic lupus erythematosus patients: the relevance of considering homologous chromosomes in determining susceptibility. AB - The aim of the present study was to determine the relevant major histocompatibility complex (MHC) class II alleles in the genetic susceptibility to systemic lupus erythematosus (SLE) in Mexican Mestizo patients. We examined the gene and haplotype frequencies of the HLA-DRB1, DQA1 and DQB1 alleles by polymerase chain reaction-sequence-specific oligonucleotide probes in 81 Mexican SLE Mestizo patients and 99 ethnically matched controls. We found a significantly increased frequency of the HLA-DRB1*0301 (p(c) = 0.031, odds ratio = 2.63) allele and significantly decreased frequencies of the DRB1*0802 (p(c) = 0.035) and DRB1*1101 (p(c) = 0.037) alleles in the SLE group. Haplotype analysis showed increased frequencies of DRB1*0301-DQA1*0501-DQB1*0201 (p(c) = 0.017, odds ratio = 2.97), and decreased frequency of DRB1*0802-DQA1*0401-DQB1*0402 (p(c) = 0.034) in SLE patients. The most frequently detected haplotypes in SLE patients showed different haplotypic combinations in the homologous chromosome from those found in controls. Thus, the combinations detected in SLE patients were either not detected in the control group or infrequently found. The results suggest that the DRB1*0301 is the principal class II allele associated with the genetic susceptibility to SLE in Mexican patients and that the presence of a specific haplotype of the homologous chromosome in patients with DRB1*0407-DQA1*03 DQB1*0302 and DRB1*1501-DQA1*0102-DQB1*0602 haplotypes could have an additive effect on the susceptibility to the disease. Finally, the low frequency of the DRB1*0301 and DRB1*1501 alleles in the control population suggests that the genetic admixture between Mexican Indians and Caucasian populations was an event that could have increased the risk of Mexicans to develop SLE. PMID- 11476906 TI - Heat-shock protein 70-1 and HLA class II gene polymorphisms associated with celiac disease susceptibility in Navarra (Spain). AB - Heat shock proteins (HSP) are thought to play a role in the immune response making probable their contribution to celiac disease (CD). We studied the polymorphisms in the 5' regulatory region of the HSP70-1 gene and performed genomic HLA-DQ and -DR typing in 128 CD patients and 94 healthy controls from Navarra (Spain). The frequency of the C allele of the HSP70-1, characterized by the intermediate electrophoretic mobility of DNA, was significantly increased among CD patients (64.5% vs 37.2%. p <1 x 10(-7)). When subjects were stratified by the HLA II genotype, differences were statistically significant between DR3 negative or DR3-DQB1*02-negative CD patients and matched controls. Homozygosity for the DQB1*02 allele was present in 48.4% of CD patients and 12.8% of controls (OR = 6.4; CI:3.1 to 13.8; p <1 x 10(-7)). Similar increased risk was observed for DQB1*02/*02, DRB1*03/-, or DRB1*03/07 patients. Furthermore, those individuals expressing the classical HLA alleles in CD (DQB1*02/*02, DRB1*03/*07) who also carried the HSP70-1 CC genotype were twelve times more likely to develop the disease than the matched controls. We therefore conclude that although HSP70 1 gene does not seem to be primarily associated with CD, it might be a component of the high risk haplotype, playing a role as an additional predisposing gene for the disease. PMID- 11476907 TI - The HLA dictionary 2001: a summary of HLA-A, -B, -C, -DRB1/3/4/5, -DQB1 alleles and their association with serologically defined HLA-A, -B, -C, -DR, and -DQ antigens. AB - This report presents the serologic equivalents of 123 HLA-A, 272 HLA-B, and 155 HLA-DRB1 alleles. The equivalents cover over 64 percent of the presently identified HLA-A, -B, and -DRB1 alleles. The dictionary is an update of the one published in 1999 (Schreuder GMTh, Hurley CK, Marsh SGE, Lau M, Maiers M, Kollman C, Noreen H. The HLA dictionary 1999: a summary of HLA-A, -B, -C, -DRB1/3/4/5, DQB1 alleles and their association with serologically defined HLA-A, -B, -C, -DR and -DQ antigens. Tissue Antigens 54:407, 1999) and also includes equivalents for HLA-C, DRB3, DRB4, DRB5, and DQB1 alleles. The data summarize information obtained by the WHO Nomenclature Committee for Factors of the HLA System, the International Cell Exchange (UCLA), the National Marrow Donor Program (NMDP), and individual laboratories. In addition, a listing is provided of alleles which are expressed as antigens with serologic reaction patterns that differ from the well established HLA specificities. The equivalents provided will be useful in guiding searches for unrelated hematopoietic stem cell donors in which patients and/or potential donors are typed by either serology or DNA-based methods. These equivalents will also serve typing and matching procedures for organ transplant programs where HLA typings from donors and from recipients on waiting lists represent mixtures of serologic and molecular typings. The tables with HLA equivalents and a questionnaire for submission of serologic reaction patterns for poorly identified allelic products will also be available on the WMDA web page: www.worldmarrow.org. PMID- 11476908 TI - Molecular typing of HLA-A, -B, and DRB using a high throughput micro array format. AB - The goal of this study was to develop a DNA micro array procedure for molecular human leukocyte antigen (HLA) typing of a large number of samples. DNA was isolated from peripheral blood samples and polymerase chain reaction (PCR) amplified for HLA-A, -B, and -DRB. Amplified DNA samples were spotted on silane treated glass slides using a micro array spotter. The spotter was capable of spotting multiple slides with up to 9216 samples per slide or 2304 samples in quadruplicate. The allele specific oligo nucleotide probes for HLA-A, -B, and DRB were labeled with the fluorescent dye Cy3, while a control probe, to quantitate the total amount of PCR product in a sample, was labeled with Cy5. Each slide was hybridized with a mixture of an allele specific Cy3 probe plus the control Cy5 probe. Following hybridization and wash, the amount of probe hybridizing to each DNA sample on the slide was measured with a micro array scanner. A computer program was used for image analysis, to calculate the average Cy3/Cy5 ratios and to identify the positive and negative samples. In turn, this information was used to determine the HLA phenotype of each sample. There was very good concordance between the results obtained for all three loci using Cy labeled probes as compared with those previously obtained by chemiluminescent detection of alkaline phosphatase labeled probes. This methodology has the potential of greatly simplifying HLA molecular typing of large number of samples. PMID- 11476909 TI - Allele- and temperature-dependency of in vitro HLA class I assembly. AB - Allelic variations of in vitro HLA class I assembly have been investigated in both the absence and the presence of binding peptides by flow cytometry using human leukocyte antigen (HLA) class I alpha chains isolated by alkali treatment from cultured HLA homozygous B cells and polystyrene beads coated with anti-HLA class I alpha chain antibodies specific to the C-terminal segment (anti-HLA class I beads). The specificity of assembly was temperature dependent, while the stability of the assembled complex depended on the bound peptide. The efficiency of assembly was allele dependent and primarily ruled by the binding affinity of alpha chains with beta(2)m. Thus, an allele hierarchy could be defined for the binding of HLA-B alpha chain with beta(2)-microglobulin: B7, B18 > B35, B62 > B27, B51. Allele and temperature dependency was found in HLA class I reassembly on acid treated B cells. The HLA class I proteins, reassembled with specific single peptides, could be efficiently transferred to anti-HLA class I beads. These findings would be used to produce microspheres coupled at high surface density with oriented single-peptide loaded HLA class I molecules and also to improve the preparation efficiency of HLA class I tetramers by the use of site specific biotinylation. PMID- 11476910 TI - Introducing Chlamydophila pneumoniae: the TWAR agent Chlamydia pneumoniae in a new perspective. PMID- 11476911 TI - The use of thalidomide in chronic refractory graft versus host disease. AB - BACKGROUND: Chronic graft versus host disease (GvHD) is a serious complication of allogenic bone marrow transplantation. As prednisone and ciclosporine A are not always effective in its treatment, we investigated the value of thalidomide. METHODS: We describe the results of thalidomide in 12 patients with chronic prednison/ciclosporin A refractory GvHD seen in our clinic since 1991. A case history is given as illustration. RESULTS: Four patients had a complete remission of chronic GvHD on thalidomide; five patients showed a partial response. Of these five patients, three died eventually of respectively ongoing GvHD, pneumonia and recurrent leukemia. There were no serious side effects, except for one patient who had to stop thalidomide because of polyneuropathy. CONCLUSIONS: It is worthwhile to give thalidomide in prednison/ciclosporin A refractory chronic GvHD. Probably thalidomide should be given earlier in the course of GvHD, this needs further study. PMID- 11476912 TI - Continuous low-dose cyclophosphamide-prednisone is effective and well tolerated in patients with advanced multiple myeloma. AB - BACKGROUND: Multiple myeloma is an incurable disease and after several lines of chemotherapy, patients enter a phase in which no standard treatment options are available. The poor outlook of these patients requires mild, palliative therapy with low toxicity. Previously used regimens either require frequent hospital attendance, lack efficacy or have significant toxicity. METHODS: In the current study, daily low dose, oral cyclophosphamide (100 mg) and prednisone (10-20 mg; CP) were administered to patients with advanced myeloma. Forty-two patients with progressive disease after melphalan-based and VAD treatment were enrolled. RESULTS: Objective responses were observed in 29 of 42 (69%) patients. In responding patients, median overall survival and progression-free survival were 22.2 months and 15.0 months, respectively. In non-responders, median OS was 3.5 months only. Side-effects were limited. Cytopenia was the most frequent event (8/29) prompting dose reduction. CP had to be stopped permanently in four patients (two cytopenia, two infections). CONCLUSION: Orally administered, low dose continuous CP is a feasible, effective and well-tolerated regimen in the management of advanced multiple myeloma. PMID- 11476913 TI - Aerosolized prostacyclin in acute respiratory distress syndrome caused by Chlamydiaceae spp. pneumonia. AB - Management of respiratory failure in acute respiratory distress syndrome (ARDS) typically requires ventilatory assistance. If traditional approaches to mechanical ventilation and adjunctive measures fail to succeed in achieving adequate oxygenation, alternative measures should be considered. We describe an ARDS patient with respiratory failure caused by a severe Chlamydiaceae species community-acquired pneumonia (CAP). Aerosolized prostacyclin (PGI(2)) treatment was successfully instituted for refractory hypoxemia. PMID- 11476914 TI - Bilateral pleural effusion and a subsegmental infiltrate due to Chlamydia pneumoniae in a mechanically ventilated patient. AB - A case of Chlamydia pneumoniae infection with bilateral pleural effusion and a subsegmental pulmonary infiltrate in an intubated and mechanically ventilated critically ill patient is described. Diagnosis was made by polymerase chain reaction on both pleural effusions. PMID- 11476915 TI - New insights into the pathogenesis of systemic lupus erythematosus (SLE): the role of apoptosis. AB - In recent years disturbances in the process of apoptosis and the clearance of apoptotic cells have been put forward as fundamental in the development of autoimmunity. In this review we will discuss the data which supply evidence for this theory. We will focus on SLE as the prototype of autoimmune disease and will review both animal studies and clinical studies in SLE patients. PMID- 11476916 TI - Management of patients with severe lung injury: first, do no harm. AB - Severe acute lung injury may result from many infectious and other insults. Although the initial insult may cause overwhelming tissue damage with subsequent gas exchange impairment and risk of death, several strategies of management may also add substantial toxicity. This review focuses on damage incurred by recently identified toxic ventilator settings e.g., high inflation pressures, too low settings of positive end-expiratory pressure, and too high tidal volumes. Liberal fluid management may be another important source of toxicity. The evidence from the literature for survival advantage of lung protective and other strategies is critically reviewed. PMID- 11476917 TI - Ultrasound and the developing central nervous system. AB - The potential risk of ultrasonography resulting in adverse biologic effects is particularly important in neurosonographic applications of diagnostic ultrasound in medicine. Key issues relate to the likelihood of producing bioeffects from the level of acoustic output emitted from modern diagnostically powerful ultrasound equipment. Important elements in the assessment of risk include the sensitivity of the tissue structures under examination, the standards of practice in clinical use and the presence of biologic effects identified from laboratory experimentation or from human studies. The World Federation for Ultrasound in Medicine and Biology continues to support activities related to evaluating bioeffects and safety of ultrasound. This paper includes extracts of some of the presentations given during the latest safety meeting, a mini-symposium on "ultrasound and the developing fetal central nervous system" held in conjunction with the WFUMB Congress in May 2000. The speakers covered topics ranging from physics of estimating heating from ultrasound equipment in clinical use to actual measurement of ultrasound-induced intracranial temperature increases in animal fetuses. Finally, some practical scanning strategies were proposed to minimise risk of adverse outcome in various clinical neurosonographic practices. PMID- 11476918 TI - Estimated fetal cerebral ultrasound exposures from clinical examinations. AB - The results of a survey of worst-case in-water values for I(SPTA), acoustic power and peak negative pressure, from a wide range of machines in clinical use, have been used with simple "fixed-path" tissue models to estimate worst-case exposure values at the fetus (in situ). The distributions of the estimated in situ values are peaked more toward their low value ends than is the case for distributions of the corresponding in-water values, with the ratio of maximum to median values being higher. Although the upper tails of the derated I(SPTA) distributions are not heavily populated, they contain important instances of relatively high values, mainly for lower frequency probes. Spectral Doppler (SD) mode produces the highest in situ estimates for all three parameters, particularly for I(SPTA) values, which exceed 5000 mW cm(-2) in a third trimester minimum attenuation model, and 900 mW cm(-2) in a "typical" attenuation model. The maximum and median values of peak negative pressure do not show particularly large differences between modes for any of the attenuation models. Values up to -3 MPa are predicted for the third trimester minimum path model, and up to -1 MPa for the "typical" attenuation model. The maximum and median values of power are dependent on the mode, with the color flow imaging (CFI) and SD modes producing the largest values (approximately 200 mW in the third trimester minimum path model). In a previous publication, more accurate calculations of temperature elevations for a bone target third trimester model were calculated for some of the probes. Extrapolation of these results for different fixed attenuation models suggests that spectral Doppler exposure may produce temperature elevations of around 6 degrees C in minimum attenuation models, and over 1 degrees C in a more "typical" attenuation model. In CFI mode, a worst-case temperature elevation of approximately 1.8 degrees C is estimated for the third trimester, and 1.3 degrees C for the first and second trimesters. For B-mode, the corresponding figures are 1.4 and 1.1 degrees C, respectively. For the case of a "typical" attenuation model, worst-case temperature elevations of approximately 0.3 degrees C are predicted for both B-mode and color flow imaging modes. PMID- 11476919 TI - Intracranial temperature elevation from diagnostic ultrasound. AB - Tissues of the central nervous system are sensitive to damage by physical agents, such as heat and ultrasound. Exposure to pulsed spectral Doppler ultrasound can significantly heat biologic tissue because of the relatively high intensities used and the need to hold the beam stationary during examinations. This has significant implications for sensitive neural tissue such as that exposed during spectral Doppler flow studies of fetal cerebral vessels. Recent changes in the FDA regulation allow delivery of almost eight times higher intensity into the fetal brain by ultrasound devices that incorporate an approved real-time output display in their design. In this situation, ultrasound users are expected to assess the risk/benefit ratio based on their interpretation of equipment output displays (including the thermal index, TI) and an understanding of the significance of biologic effects. To assist in the assessment of potential thermally mediated bioeffects, a number of conclusions can be drawn from the published scientific literature: the amount of ultrasound-induced intracranial heating increases with gestational age and the development of fetal bone; pulsed spectral Doppler ultrasound can produce biologically significant heating in the fetal brain; the rate of heating near bone is rapid, with approximately 75% of the maximum heating occurring within 30 s; blood flow has minimal cooling effect on ultrasound-induced heating of the brain when insonated with narrow focused clinical beams; the threshold for irreversible damage in the developing embryo and fetal brain is exceeded when a temperature increase of 4 degrees C is maintained for 5 min; an ultrasound exposure that produces a temperature increase of up to 1.5 degrees C in 120 s does not elicit measurable electrophysiologic responses in fetal brain; for some exposure conditions, the thermal index (TI), as used in the FDA-approved output display standard, underestimates the extent of ultrasound-induced intracranial temperature increase. PMID- 11476920 TI - Clinical concerns in the ultrasound exposure of the developing central nervous system. AB - Neurosonography is used as a primary imaging modality worldwide for visualization of the developing brain and spinal cord in fetuses, infants and children. During the entire process of brain development, there is rapid cell turnover, a condition that is favorable for genetic mutations once external stimuli are applied. No clinical studies in humans have been performed specifically to discuss the long-term impact of postnatal ultrasound exposure of the central nervous system. Currently published studies concerning the prenatal and postnatal use of Doppler or of ultrasound contrast agent use and the developing central nervous system are insufficient to draw meaningful conclusions regarding safety. By instituting a standardized examination and following appropriate patient handling guidelines, the risk of an adverse outcome associated with neurosonography is minimized. This paper recommends adoption of the ALARA principle and offers suggestions as to how to minimize the risk of adverse effects in neurosonography. PMID- 11476921 TI - Value of splanchnic Doppler ultrasound in the diagnosis of portal hypertension. AB - The accuracy of various Doppler parameters of portal circulation in the diagnosis of relevant portal hypertension (presence of gastroesophageal varices) was prospectively validated. The following parameters were compared in 51 patients with chronic liver disease (40 with cirrhosis and 11 with chronic hepatitis): portal vein flow velocity and congestion index, hepatic and splenic arteries resistance indexes (RI), modified liver vascular index (portal flow velocity/hepatic artery RI) and portal hypertension index, a new index calculated as: [(hepatic artery RI x 0.69) x (splenic artery RI x 0.87)]/portal vein flow velocity. Highest accuracy was achieved by the splenic artery RI and the portal hypertension index (both around 75%) at cut-offs, respectively, of 0.60 and 12 cm/s(-1), which appeared to be, therefore, the most favorable parameters for the clinical practice. Their use may limit the need for endoscopy to search for varices. PMID- 11476922 TI - Noninvasive grading of the severity of portal hypertension in cirrhotic patients by echo-color-Doppler. AB - The possibility of estimating portal hypertension combining splenic Doppler pulsatility index (PI), which has been shown strictly to be related to portal resistance, and portal blood flow (PBF) was evaluated. Hepatic venous pressure gradient, and splanchnic Doppler parameters were measured in 40 cirrhotic patients. A formula to assess the severity of portal hypertension from Doppler parameters was calculated in a training group of 19 patients, and then this formula was tested to prospectively predict the degree of portal pressure in 21 further patients. In the training group, the regression of portal resistance over splenic PI was calculated and individual values of estimated portal resistance were obtained for every patient of the test group. From them and from observed values of PBF, an estimated value of portal pressure was calculated. The following formula was obtained [(0.066*splenic PI -0.044)*PBF]. In the second group, predicted and observed portal pressure were well correlated (r = 0.71, p = .0003). A good accuracy for the prediction of mild or severe portal hypertension was obtained. PMID- 11476923 TI - Contrast-enhanced power Doppler imaging of normal and decreased blood flow in canine prostates. AB - The purpose of this study was to investigate if Sonazoid, a new ultrasound (US) contrast agent, can improve the delineation of areas with normal and decreased blood flow in the prostate. Sonazoid was administered in the dose range of 0.00625-0.0375 microL microbubbles/kg into five anaesthetised mongrel adult male dogs. Transrectal power Doppler imaging of the prostate was performed in 2-D and 3-D with a C9-5 end-fire probe, using an HDI 3000 scanner. An area of decreased blood flow was created by inducing tissue ablation with a CL60 laser system, to mimic an avascular lesion. A subjective assessment of the intraprostatic vessels and the prostate vascular architecture was performed, with and without Sonazoid, before and after inducing the abnormal site. Visibility of the prostate blood flow improved following Sonazoid injection (p < 0.001). A symmetric, radial vascular pattern was identified in the normal prostate prior to tissue ablation, but only on the enhanced images. After tissue ablation, a disturbance of the normal vascular pattern and identification of areas with lack of blood flow was possible, following Sonazoid injection. Furthermore, the location and size of these areas were verified in all dogs by gross histology examination. Sonazoid enhances the visibility of the prostate vascular architecture and improves, thereby the delineation of areas with normal and decreased blood flow. PMID- 11476924 TI - Harmonic imaging of the brain parenchyma using a perfluorobutane-containing ultrasound contrast agent. AB - We evaluated the signal-enhancing effect of the novel perfluorobutane-based ultrasound contrast agent BR 14 (Bracco Research, Switzerland) in grey-scale harmonic imaging of the brain parenchyma. Six sedated male beagle dogs were investigated with transcranial grey-scale harmonic imaging (SONOS 5500, 1.8/3.6 MHz). After bolus injection of two different doses of BR 14, acoustic densitometry was performed to quantify changes in regional contrast intensity. In the dogs' brain parenchyma, the mean relative peak increase in acoustic intensity was +61% after administration of 0.05 ml/kg BW of BR 14 and +24% after 0.2 ml/kg BW. In the masticatory muscle, application of the higher dose resulted in a stronger increase in contrast intensity compared to the lower dose. Evaluation of the contralateral base of the skull showed a dose-dependent decrease in acoustic intensity. Bolus injection of BR 14 produces an increase in acoustic intensity, which can be used for the visualization of contrast agent in the brain parenchyma. Using high dosages, a strong signal-enhancing effect in the regions near the ultrasound probe leads to a consecutive attenuation of signals from structures being located beyond ("shadowing-effect"). This is the explanation for the paradoxical result that the higher dose leads to a lower peak signal increase in the brain parenchyma. PMID- 11476925 TI - Dorsal aortic flow velocity in chick embryos of stage 16 to 28. AB - The objective of this study was to evaluate two Doppler frequency-detection methods to measure blood flow velocity in the developing chick embryo. We compared the commonly used directional zero-crossing counter and a customized digital bidirectional spectrum analyzer. At development stages 16 up to 28 (2.5 to 6 days incubation), a reversed flow component in the dorsal aorta was demonstrated using the bidirectional spectrum analyzer. Dorsal aortic velocities obtained with the directional zero-crossing counter were significantly lower than with the bidirectional spectrum analyzer in stages 16, 20 and 28. In addition to the differences in the absolute velocity values, there was also a remarkable discrepancy in the velocity waveform shape using the two Doppler frequency processors. The calculated heart rate using the two Doppler frequency processors was identical. It is concluded that a Doppler velocity detector based on spectral analysis is superior to the hitherto used zero-crossing counter in the chick embryo. With the customized digital bidirectional spectrum analyzer, we can accurately measure the hemodynamics of the developing chick embryo. PMID- 11476926 TI - Accuracy of biphasic response, sustained improvement and worsening during dobutamine echocardiography in predicting recovery of resting myocardial dysfunction after revascularization: comparison with thallium-201 SPECT. AB - To evaluate the accuracy of various types of wall motion response during dobutamine echocardiography (DE) in predicting functional recovery after revascularization, we studied 30 patients with stable coronary disease and left ventricular dysfunction by simultaneous DE and (201)Tl reinjection SPECT. Among 480 segments (16 segments/patient), 199 had abnormal wall motion at baseline and 167 were revascularized. The predictive value for recovery of function was 72% for a biphasic response, 61% for sustained improvement, 77% for worsening, and 27% for no change (p < 0.01 vs. each). Biphasic response had a sensitivity of 40% and specificity of 85%. Combining biphasic, sustained improvement and worsening responses, the sensitivity, specificity and accuracy were 76%, 65% and 71%, respectively. For (201)Tl SPECT, they were 90%, 65% and 78%, respectively. Thus, a biphasic response alone is of low sensitivity. Combination of biphasic, sustained improvement and worsening responses gives an accuracy rate comparable to that of (201)Tl reinjection SPECT in assessing functional recovery. PMID- 11476927 TI - Brain microembolic counts during extracorporeal circulation depend on aortic cannula position. AB - Thrombofibrinous and gaseous brain microemboli are commonly observed during extracorporeal circulation in patients undergoing cardiac surgery. Recent studies showed a preference of microemboli to the left hemisphere. We hypothesized that placement of the aortic cannula tip within the aorta descendens rather than in the aorta ascendens as commonly used reduces the number of microemboli to the brain and also reduces the side preference. Of 60 patients undergoing elective coronary artery bypass grafting primarily randomized to either a short aortic cannula (aorta ascendens) or an elongated one (aorta descendens), 29 (16 and 13) patients had complete and technically perfect continuous intraoperative transcranial Doppler sonography with embolus detection bilaterally. The number of high-intensity transient signals (HITS) was 994 +/- 2118 (mean +/- SD) for the short cannula group and was significantly lower with the elongated cannula (223 +/- 208; p < 0.02). HITS counts per min also differed significantly in favour of the aorta descendens group (p < 0.02), but there was an overlap. Thus, elongated cannulas can reduce, but not prevent microembolism to the brain. The side-to-side ratio of microemboli revealed more events in the left hemisphere, but this was similar in both groups. This suggests that individual anatomic factors may be responsible for this hemodynamically-mediated effect. PMID- 11476928 TI - Feasibility of the flash-replenishment concept in renal tissue: which parameters affect the assessment of the contrast replenishment? AB - The purpose of the study was to evaluate whether power pulse inversion (PPI) and pulse inversion (PI) techniques allow the measurement of indices of microcirculatory flow in real-time at low emission power using contrast microbubbles. PPI and PI imaging were performed in a kidney perfusion model during continuous infusion of Definity (0.12 mL/min). At steady state of tissue enhancement, contrast was destroyed by emission of echo bursts at high emission power (MI = 1.3). Consecutively, contrast replenishment was assessed at low emission power (MI = 0.09) in real-time imaging modes (PPI: 12 Hz; PI: 25 Hz). Regions-of-interest (ROI) of variable sizes were placed in the renal cortex and bigger arteries to compare replenishment of macro- and microcirculation. Nonlinear curve fitting was performed using the mathematical model y=s+A(1-e( betat)), with A as the parameter describing blood volume and beta as a parameter describing the speed of microbubble contrast replenishment. Replenishment curves could be visually appreciated and quantitatively analyzed in all renal segments. A was significantly higher in bigger arteries compared to renal cortex (p < 0.001). beta was found to be significantly higher in the arteries as compared to the cortex (p < 0.001). The SD of beta diminishes with increasing size of the ROI. The acquisition of replenishment curves following ultrasound (US)-induced destruction of contrast microbubbles is feasible at low power using PPI and PI. Assessment of replenishment kinetics allows the differentiation between macro- and microcirculation. Size and position of the ROI have an important impact on the generation of replenishment curves in both imaging modalities, which has to be taken into account. PMID- 11476929 TI - Evaluation of voxel-based registration of 3-D power Doppler ultrasound and 3-D magnetic resonance angiographic images of carotid arteries. AB - Spatial registration and fusion of ultrasound (US) images with other modalities may aid clinical interpretation. We implemented and evaluated on patient data an automated retrospective registration of magnetic resonance angiography (MRA) carotid bifurcation images with 3-D power Doppler ultrasound (PD US) and indirectly with 3-D B-mode US. Volumes were initially thresholded to reduce the uncorrelated noise signals. The registration algorithm subsequently maximized the mutual information measure between the PD US and 3-D MRA via iterative simplex search to find best "rigid body" transformation. We rated the performance of the algorithm visually on (n = 5) clinical MRA and 3-D PD US datasets. We also evaluated quantitatively the effect of thresholding, initial misalignment of the paired volumes and the reproducibility registration. We investigated the effect of image artefacts by simulation experiments. Preregistration misalignments of up to 5 mm in the transaxial plane, up to 10 mm along the axis of the carotids and up to 40 degrees resulted in 107 of 110 successful registrations, with translational and rotational errors of 0.32 mm +/- 0.3 mm and 1.6 +/- 2.1 degrees. The algorithm was not affected by missing arterial segments of up to 8 mm in length. The average registration time was 4 min. We conclude that the algorithm could be applied to 3-D US PD and MRA data for automated multimodality registration of carotid vessels without the use of fiducials. PMID- 11476930 TI - Optimisation and evaluation of an electromagnetic tracking device for high accuracy three-dimensional ultrasound imaging of the carotid arteries. AB - Electromagnetic tracking devices provide a flexible, low cost solution for three dimensional ultrasound (3-D US) imaging. They are, however, susceptible to interference. A commercial device (Ascension pcBIRD) was evaluated to assess the accuracy in locating the scan probe as part of a digital, freehand 3-D US imaging system aimed at vascular applications. The device was optimised by selecting a measurement rate and filter setting that minimised the mean deviation in repeated position and orientation measurements. Experimental evaluation of accuracy indicated that, overall, absolute errors were small: the RMS absolute error was 0.2 mm (range: -0.7 to 0.5 mm) for positional measurements over translations up to 90 mm, and 0.2 degrees (range: -0.8 to 0.9 degrees ) for rotational measurements up to 30 degrees. In the case of position measurements, the absolute errors were influenced by the location of the scanner relative to the scan volume. We conclude that the device tested provides an accuracy sufficient for use within a freehand 3-D US system for carotid artery imaging. PMID- 11476931 TI - Three-dimensional forward-viewing intravascular ultrasound imaging of human arteries in vitro. AB - The aim of this work was to investigate the suitability of a novel forward viewing intravascular ultrasound (IVUS) technique for three-dimensional imaging of severely stenosed or totally occluded vessels, where the conventional side viewing IVUS systems are of limited use. A stiff 3.8 mm diameter forward-viewing catheter was manufactured to scan a 72 degrees sector ahead of its tip. Conical volume data were acquired by rotating the catheter over 180 degrees by means of a motorised mechanical system. Operating at 30 MHz, the catheter was integrated with an IVUS scanner and a radiofrequency data acquisition system. Postmortem carotid and femoral arteries were scanned in vitro. Correlation of the reconstructed images with histology demonstrated the ability of this forward viewing IVUS system to visualise healthy lumens, bifurcations, thickened atherosclerotic walls and, most importantly, severe and complete vessel occlusions. A rotating-sector forward-viewing IVUS system is suitable for anatomical assessment of severely diseased vessels in three dimensions. PMID- 11476932 TI - Ultrasonic characteristics of leiomyoma uteri in vitro. AB - Patients suffer with various abnormalities of body tissues. "Leiomyoma Uteri" is a solid tumor of uterus, one of such abnormalities. To treat such tumors, basic physical and biologic investigations are required to be carried out. In the present work, ultrasonic characterization of soft tissues, in this case, uterine tumor in vitro, are studied. A double probe through-transmission technique is used for the measurement of these propagation parameters, viz., velocity and attenuation. Other parameters like acoustic impedance, dynamic modulus of elasticity and compressibility are also determined. The average velocity and attenuation are found to be 1550 ms(-1) and 433 dBm(-1), respectively, at 3.5 MHz frequency and room temperature 28 degrees C. The present investigation is useful in tissue differentiation and tissue identification to enable the doctors to give proper treatment. PMID- 11476933 TI - Skeletal effects of low-intensity pulsed ultrasound on the ovariectomized rodent. AB - Growing evidence supports low-intensity pulsed ultrasound (US) as an osteogenic mechanical stimulus. Its effects on isolated bone cells and on fractured bone are established. However, its effects on osteoporosis are not clear. This study examined US effects on ovariectomy (OVX) induced bone changes within the rodent hindlimb (distal femur and proximal tibia), and on normal bone in animals following sham-OVX. Animals were exposed to daily unilateral active-US and contralateral inactive-US for 12 weeks. Bone status was assessed using dual energy X-ray absorptiometry and histomorphometry. Ovariectomy resulted in significant bone changes. Low-intensity pulsed US did not influence these changes. These results suggest that the US dose introduced may not be a beneficial treatment for osteoporosis, and that intact bone may be less sensitive to US than fractured bone and isolated bone cells. This may relate to the biophysical mechanisms of action of US, US-bone interactions and tissue level processes taking place. PMID- 11476934 TI - Effects of pulsed ultrasound on development and glucose uptake of preimplantation mouse embryos. AB - The effects of pulsed ultrasound (US) on early mouse embryos were investigated. Two-cell embryos contained in oviducts were irradiated to US (1.875 MHz with an I (SPTA) of 2.96 W/cm(2)) in an experimental unit for either 1 or 5 min (exposure groups). The embryos were cultured to examine the rate of developing to blastocysts, and the uptake of 2-deoxyglucose (2-DG) into blastocysts was measured to evaluate their viability. The rates in the exposure groups were essentially the same, with those of the embryos treated similarly in the unit unless being exposed to US (nonexposure groups). However, they were lower than that of embryos not treated in the experimental unit (a control group). There were no significant differences of 2-DG uptake among the 1-min exposure, 1-min nonexposure, and control groups. The uptake in the 5-min exposure group did not differ from that in the 5-min nonexposure group; however, uptake in both groups was lower than that in the control group. Pulsed US for 1 min did not affect viability of preimplantation mouse embryos. PMID- 11476935 TI - Low-reflection-coefficient liquid interfaces for system characterization. AB - The use of liquid brominated hydrocarbons to form a planar reflecting interface with water is described. Gravity-based planar reflecting surfaces with known reflection coefficients can be used in system characterization for quantitative ultrasonics, and a set of surfaces with a range of reflection coefficients allows calibration of the output power and receiver gain of ultrasonic imaging systems. The substances reported here are immiscible in water and form interfaces with water, resulting in a broad range of acoustic reflection coefficients. Reflection coefficients were measured at temperatures from 18-24 degrees C for "pure" substances and for mixtures of two brominated hydrocarbons. Results show that reflection coefficients are weakly dependent on temperature and that, at a specific temperature, a significant range of arbitrarily small reflection coefficients is available, in the case of the mixtures, by the appropriate choice of weight-percents of the two brominated hydrocarbons. PMID- 11476936 TI - Woc (without children) gene control of ecdysone biosynthesis in Drosophila melanogaster. AB - The first step in ecdysteroidogenesis, i.e. the 7,8-dehydrogenation of dietary cholesterol (C) to 7-dehydrocholesterol (7dC), is blocked in Drosophila melanogaster homozygous woc (without children) third instar larval ring glands (source of ecdysone). Unlike ring glands from wild-type D. melanogaster larvae, glands from woc mutants cannot convert radiolabelled C or 25-hydroxycholesterol (25C) to 7dC or 7-dehydro-25-hydroxycholesterol (7d25C) in vitro, nor to ecdysone (E). Yet, when these same glands are incubated with synthetic tracer 7d25C, the rate of metabolism of this polar Delta(5,7)-sterol into E is identical to that observed with glands from comparably staged wild-type larvae. The absence of this enzymatic activity in vivo is probably the direct cause of the observed low whole body ecdysteroid titers in late third instar homozygous mutant larvae, the low ecdysteroid secretory activity in vitro of brain-ring gland complexes from these animals, and the failure of the larvae to pupariate (undergo metamorphosis). Oral administration of 7dC, but not C, results in a dramatic increase in ecdysteroid production both in vivo and in vitro by the woc mutant brain-ring gland complexes and affects a partial rescue to the beginning of pupal-adult development, but no further, despite elevated whole-body ecdysteroid titers. Data previously reported (Wismar et al., 2000) indicate that the woc gene encodes a zinc-finger protein that apparently modulates the activity of the 7,8-dehydrogenase. PMID- 11476937 TI - Human gonadotropin-releasing hormone receptor gene transcription: up-regulation by 3',5'-cyclic adenosine monophosphate/protein kinase A pathway. AB - Transient transfection of mouse gonadotrope-derived (alphaT3-1) cells with a 2297 bp human GnRHR promoter-luciferase construct (p2300-LucF) showed a dose- and time dependent increase in the human gonodotropin-releasing hormone receptor (GnRHR) promoter activity after forskolin treatment. An average of 4.8-fold increase in promoter activity was observed after 12 h of 10 microM forskolin treatment. This effect was mimicked by administration of cholera toxin, cAMP analog or pituitary adenylate cyclase activating polypeptide 38 (PACAP). A specific adenylate cyclase (AC) inhibitor (ACI) or protein kinase A (PKA) inhibitor (PKAI) pretreatment reversed the forskolin- and PACAP-induced increase in the human GnRHR promoter activity. These results not only confirm the stimulatory effect of Cyclic adenosine monophosphate (cAMP) in human GnRHR promoter activation, but also suggest that hormones or neurotransmitters that activate adenylate cyclase in pituitary gonadotropes may increase the expression of human GnRHR gene in transcriptional level. Progressive 5' deletion assays identified a 412 bp fragment (-577 to 167) in the human GnRHR 5'-flanking region that is essential in maintaining the basal responsiveness to cAMP. Mutagenesis coupled with functional studies have identified two putative AP-1/CREB binding sites, namely hGR-AP/CRE-1 and hGR-AP/CRE-2 that participated in mediating the cAMP-stimulatory effect. Mutation of the putative hGR-AP/CRE-1 and hGR-CRE-2 resulted in a 38 and 32% decrease in the forskolin-induced stimulation. However, mutation of both binding sites did not completely abolish the cAMP-stimulatory effect, suggesting that multiple transcription factor binding sites were involved in full response in cAMP stimulation. The binding of CREB to these motifs was confirmed by gel mobility shift assay and antibody supershift assay. PMID- 11476938 TI - Characterization of domains in C/EBPalpha that mediate its constitutive and cAMP inducible activities. AB - Structure/function analysis of CCAAT/enhancer binding proteins (C/EBP) alpha and beta have shown that they possess both constitutive and cAMP inducible activities. Three regions conserved between C/EBPalpha and beta were identified which lie within the cAMP inducible domains of each protein. Deletion analysis of these conserved regions within C/EBPalpha show that conserved region 2 plays a particularly critical role in mediating the PKA inducible activity of the protein, however, the constitutive activity of conserved region 2 depends on promoter context. This data supports previous findings that constitutive and cAMP responsiveness are mediated by domains of the protein that do not directly overlap, suggesting that they occur through distinct mechanisms. PMID- 11476939 TI - Selective insulin-induced activation of class I(A) phosphoinositide 3-kinase in PIKfyve immune complexes from 3T3-L1 adipocytes. AB - A diverse range of insulin-regulated cellular processes are dependent on class I(A) phosphatidylinositol 3-kinases (PI 3-Ks) and their association with and activation by up-stream signaling molecules. Here we report on the identification of the phosphoinositide 5'-kinase PIKfyve as a partner of class I(A) PI 3-K. Thus, both p85 and p110 subunits (class I(A)) of PI 3-Ks co-precipitated with anti-PIKfyve antibodies from lysates of resting 3T3-L1 adipocytes and, vice versa, PIKfyve co-precipitated with anti-p85 PI 3-K antibodies. Assignment to class I(A) PI 3-K enzymatic activity was further substantiated by the inhibition of PtdIns 3-P production in PIKfyve immune complexes by low concentrations of wortmannin and Triton X-100, and its preferences for Mg(2+) versus Mn(2+). Insulin but not PDGF or EGF stimulation of 3T3-L1 adipocytes markedly increased the PtdIns 3-P production (4.2-fold) in PIKfyve immune complexes, primarily as a result of increased PI 3-K intrinsic enzymatic activity. Intriguingly, while both insulin and PDGF caused an increase of class I(A) PI 3-K activity co immunoprecipitated with tyrosine phosphorylated proteins, only insulin treatment yielded an activation of class I(A) PI 3-K in PIKfyve immune complexes. Studies aiming at identifying the underlying mechanism revealed that PIKfyve-class I(A) PI 3-K association and the insulin-induced activation likely operate independently of tyrosine phosphorylated insulin receptor substrate proteins. Together, these results establish PIKfyve as a novel source of activated class I(A) PI 3-K molecules that may be relevant in the insulin-signal transduction pathway. PMID- 11476940 TI - Paracrine action of keratinocyte growth factor (KGF) during ruminant mammogenesis. AB - Keratinocyte growth factor (KGF) is a stroma-derived mitogen mediating epithelial stromal interactions. We investigated the role of KGF during epithelial-stromal interactions accompanying ruminant mammogenesis. Target-specificity of KGF was demonstrated in that KGF-stimulated proliferation of bovine mammary epithelial, but not ovine mammary stromal cells. Consistent with a paracrine function, 4.6, 2.4, 1.5 and 0.9 kb mRNA transcripts were expressed by bovine stromal, but not epithelial cells. Within the ovine mammary gland, 2.4 and 1.5 kb KGF mRNAs were expressed in the fat pad while only the 2.4 kb transcript was transcribed in parenchyma. The abundance of KGF mRNA was greater in the extra-parenchymal fat pad than in the contralateral epithelium-free fat pad prior to puberty, and was less in parenchyma than in the intact or epithelium-free fat pads. Ovariectomy tended to increase KGF transcription while estrogen reduced expression. Of several tissues, mammary parenchyma expressed a 2.4 kb mRNA while adipose tissues expressed a 1.5 kb transcript. These results demonstrate local and systemic regulation of KGF transcription and support a paracrine role for KGF during ruminant mammogenesis. PMID- 11476941 TI - Kap promoter analysis in vivo: a regulatory role for a truncated L1 repeat. AB - Reporter gene expression directed by a 1542-base pair (bp) fragment of the Kap promoter is specific to the proximal tubules of the kidney and androgen regulated. In the present study, the characteristics of the androgen response from the 1542-bp promoter were examined in vivo. The estrogen response in the kidney and uterus was also examined. The reporter gene expression was assayed in lines of transgenic mice generated from a truncated promoter construct in which the L1 repeat, present at the distal portion of the 1542-bp, had been deleted. The pattern of androgen response of the reporter gene is similar to that of the endogenous Kap. Reporter gene expression in the 1542-bp promoter does not respond to estrogen in the kidney, while perinatal expression in the uterus does occur. Truncation of the L1 results in loss of reporter gene expression. We conclude that L1 sequences present near the Kap promoter have a regulatory function. PMID- 11476942 TI - Calcitonin is a prostate epithelium-derived growth stimulatory peptide. AB - Locally secreted growth factors and neuropeptides may play an important role in sustaining the growth of hormone-independent prostate cancer. Our previous studies have shown that calcitonin-like immunoreactive peptide (CTI) is secreted by primary prostate cells in culture, and its secretion from malignant prostate cells is significantly higher than benign cells. Exogenously added calcitonin (CT) induces DNA synthesis in serum-starved prostate cancer LNCaP and PC-3M cells. Present studies extended these findings by cloning cDNAs for CT and CT receptor (CT-R) from prostate cancer cells and studying the expression of CT and CT-R mRNA in prostate cancer cell lines and primary prostate tumor specimens. The results have shown that PC-3 cells expressed CT, and not CT-R, mRNA, whereas CT R, but not CT, mRNA was expressed by LNCaP cells. Conditioned media from PC-3 cells induced DNA synthesis of LNCaP cells, and this mitogenic response was abolished by anti-CT serum. Highly aggressive PC-3M cells co-expressed CT and CT R mRNAs. CT also induced a twofold increase in DNA synthesis of primary prostate cells and anti-CT serum caused a 56% decline. In-situ hybridization histochemistry of archival prostate specimens has selectively localized CT and CT R mRNA in basal epithelium of benign and low grade PC specimens, and these mRNAs were not detected in either luminal epithelium or stroma. In contrast, CT and CT R mRNA were detected throughout the luminal epithelium of moderate and high-grade PC specimens. Most epithelial cells of low and moderately differentiated tumors expressed either CT or CT-R mRNA, suggesting that CT may serve as a paracrine factor. In contrast, CT and CT-R mRNAs were co-expressed by most tumor cells in advanced PC specimens. The cells expressing CT-R mRNA in primary tumors also co expressed PCNA. These results, when combined with mitogenic actions of CT on primary prostate cells as well as PC cell lines, strongly support the role for CT in sustaining the growth of cancer cells. PMID- 11476943 TI - Increased expression of the Zn-finger transcription factor BTEB1 in human endometrial cells is correlated with distinct cell phenotype, gene expression patterns, and proliferative responsiveness to serum and TGF-beta1. AB - Basic transcription element binding (BTEB, also designated BTEB1) protein is a member of the Sp-family of GC-box binding transcription factors that exhibit distinct patterns of expression in many cell types and tissues. A role for BTEB1 in the regulation of cell growth and gene transcription has been invoked, but little is known about the molecular mechanisms underlying these activities. The present study examined the functional consequences of high and low BTEB1 expression in the human endometrial carcinoma cell line Hec-1-A, by deriving stable clonal lines that expressed sense (S) and anti-sense (As) rat BTEB1 constructs. Clonal S lines, with BTEB1 mRNA and protein levels higher than in corresponding parent (N) and As lines, displayed enhanced DNA synthesis upon 3[H] thymidine incorporation, in serum-containing but not in serum-free medium, and increased cell cycle kinetics, concomitant with the induction in expression of the genes for the cell cycle-associated components cyclin D1, PCNA, cyclin dependent kinase (Cdk) inhibitor p21, and Cdk2. Compared to N and As lines, S lines also had diminished ability to grow in multi-layers and exhibited increased mRNA levels for plasminogen activator inhibitor-1 (PAI-1), secretory leukocyte protease inhibitor (SLPI), and tissue inhibitor of metalloproteinases (TIMP)-2. In serum-free medium, S, but not N nor As lines, had enhanced DNA synthesis with transforming growth factor (TGF)-beta1, albeit all lines demonstrated similar responses to insulin-like growth factor-I and to epidermal growth factor, respectively. The higher DNA synthesis in S relative to N and As, lines upon exogenous TGF-beta1 addition, was observed in concert with increased expression of cyclins D1 and E and p21, genes. Moreover, S and As lines had increased mRNA levels for TIMP-1, TIMP-2, PAI-1, and beta-catenin, and diminished SLPI, and to a lesser extent, Cdk4 mRNA levels, with TGF-beta1 treatment. These results suggest that BTEB1 may mediate cell growth, in part, by modulating gene expression levels of distinct cell cycle and growth-associated proteins. The correlation between serum- and TGF-beta1 induction of DNA synthesis with increased BTEB1 expression further suggests that BTEB1 may constitute an important downstream regulatory component of various signaling pathways utilized by serum-associated and other growth factors in endometrial epithelial cells. PMID- 11476944 TI - Expression of thyroid hormone receptor betaA gene assayed by transgenic Xenopus laevis carrying its promoter sequences. AB - The responsiveness of thyroid hormone responsive element (TRE)-containing promoter sequence to thyroid hormone (TH) was studied utilizing Xenopus laevis carrying a transgene containing 5'-upstream region of TRbetaA1 gene and green fluorescent protein (EGFP) gene. EGFP-expression was seen first in neurulae, which continued to stage 45, then became weak, and again started to increase at the prometamorphic stage, culminating at the metamorphic climax stage. Immunohistochemistry identified eyes, viscera, and muscles as the EGFP-expressing larval tissues. The treatment of premetamorphic tadpoles with TH induced the precocious EGFP-expression. We also showed that the transgenic Xenopus adults were responsive to exogenous TH, a high responsiveness being seen in brain, small intestine, kidney, and bone. TRbeta-expression in the embryo, larva, and adult was verified by Western blotting. Thus, TH not only regulates the metamorphosis, but also might play some biological role(s) in embryos and adults. PMID- 11476945 TI - Virilization of the urogenital sinus of the tammar wallaby is not unique to 5alpha-androstane-3alpha,17beta-diol. AB - The androgen 5alpha-androstane-3alpha,17beta-diol (5alpha-adiol) is synthesized in testes and secreted into plasma of male tammar wallaby pouch young and appears to virilize the urogenital sinus. To provide insight into its mechanism of action, a dose response study showed that administration of 1 microg 5alpha-adiol monoenanthate per g body wt. per week for 3 weeks to 24-day-old female pouch young induced prostate bud formation equivalent to that of males of the same age. Administration of this same dose of the enanthates of testosterone, dihydrotestosterone, and 5alpha-adiol to female pouch young caused equivalent virilization of the urogenital sinus. The fact that 5alpha-adiol does not exert a unique effect, together with our earlier findings in this species that 5alpha adiol and testosterone are converted to dihydrotestosterone in the urogenital sinus and that virilization of the urogenital sinus is prevented by the androgen receptor antagonist flutamide, suggest that 5alpha-adiol is a circulating precursor for dihydrotestosterone formation in this tissue. PMID- 11476946 TI - Differential cellular localization of estrogen receptor alpha in uterine and mammary cells. AB - The classical alpha isoform of the estrogen receptor (ER) has been reported to localize almost exclusively in the nucleus. However, studies on non-genomic steroid effects have also suggested the existence of ERs residing at the cell surface. In this work, we present immunological data supporting extra-nuclear ERalpha localization in uterine (SHM) and mammary (MCF-7) cell lines. Immunocytological studies performed on SHM cells revealed that native ERs mainly localize as a perinuclear cytoplasmic ring. The receptors were rapidly translocated to the nucleus by 17beta-estradiol. In addition to nuclear ERs, a peripheral reservoir of ERalpha immunoreactivity, most probably associated with the plasma membrane, was detected in MCF-7 cells. These results were confirmed by the detection of membrane estrogen binding sites using fluorescent estrogen-BSA derivatives and ligand binding assays to intact cells, where [3H]-estradiol could be partly displaced by impeded estrogen conjugates. Partial inhibition of radioligand binding by an antibody against the steroid binding domain of the ERalpha suggests that the isoform faces the extracellular media in MCF-7 cells. Moreover, ERalpha-like proteins ( approximately 67 kDa) were found to be associated in isolated membrane subfractions from the cells. However, immunocytology of COS-1 (ER-negative) and SHM cells transfected with the complete cDNA coding for the cloned ERalpha and beta isoforms showed exclusive nuclear localization of the overexpressed ERs. The non-classical distribution of native ERalpha-like proteins in each cell line, suggests an alternative mode of ERalpha cellular localization/function. Cell type-dependent processing may account for the differential localization shown by native and expressed receptors in the systems considered. PMID- 11476947 TI - T3-activation of the rat growth hormone gene is inhibited by a zinc finger/homeodomain protein. AB - Since the transcription factor Zfhep is expressed in somatotropes and binds the rat growth hormone (rGH) gene T3-response element (TRE), we investigated whether Zfhep regulates the response of this gene to T3. In cotransfection experiments, Zfhep did not regulate the native rGH promoter in the absence of T3. However, Zfhep repressed T3-mediated activation significantly in either GH(3) or JEG-3 cells. Up to 70% repression was mediated through the rGH TRE in a heterologous promoter (thymidine kinase), but was not observed with the idealized DR4 or chicken lysozyme F2 TREs. Zfhep apparently does not repress T3-mediated activation simply by competition for binding to DNA since the C-terminal DNA binding domain of Zfhep (which is sufficient for DNA-binding) is not sufficient for repression and since cotransfection of excess thyroid hormone receptor (TR) did not prevent repression by Zfhep. These data indicate that the rGH TRE is a composite element that can integrate Zfhep and T3 regulation. PMID- 11476948 TI - Human estrogen receptor beta-specific monoclonal antibodies: characterization and use in studies of estrogen receptor beta protein expression in reproductive tissues. AB - Investigation of the role of the second, more recently described estrogen receptor, denoted ERbeta, will be critical in understanding the molecular mechanisms underlying tissue-specific gene regulation by estrogens. Expression of ERbeta in a variety of tissues has been examined predominantly at the mRNA level, and there is little information regarding the cellular localization and size of the endogenous ERbeta protein, due, in part, to the limited availability of human ERbeta-specific antibodies. Thus, our aim was to generate specific antibodies to human ERbeta and use them to determine the tissue-specific distribution and size(s) of the ERbeta protein. To this end, we have cloned three different hybridoma cell lines that produce monoclonal antibodies specific for the hormone binding domain of human ERbeta. The antibodies, made in mice against human ERbeta amino acids 256-505 (hormone binding domain lacking the F domain), are designated CFK-E12 (E12), CMK-A9 (A9) and CWK-F12 (F12) and were determined to be the IgG gamma1 isotype for E12, and IgG gamma2b for A9 and F12. All three monoclonal antibodies could be used to detect in vitro translated, baculovirus expressed, and cell transfected and expressed ERbeta protein by Western blot analyses, and all failed to detect ERalpha. A9 and F12 were able to immunoprecipitate efficiently the native form of ERbeta protein in the presence and absence of estradiol. Epitope mapping studies indicate that the E12 and F12 antibodies recognize overlapping peptide sequences in the N-terminal region of the hormone binding domain, a region that is highly conserved among species. Immunocytochemical studies with these antibodies reveal nuclear-specific localization of the ERbeta protein in granulosa cells of the rat ovary. Nuclear ERbeta is also specifically localized in epithelial and some stromal cells of mouse and rat epididymis. Western blot analysis with protein extracts from ovarian granulosa cells of human, rat, mouse, and pig showed a ca. 52 kDa and an additional ca. 62-64 kDa band in these species. These results indicate the presence of two predominant molecular size forms of the ERbeta protein in ovarian granulosa cells and demonstrate the utility of these antibodies for detection of ERbeta in the human and in several other mammalian species. PMID- 11476949 TI - Interaction of estrogen receptors alpha and beta with estrogen response elements. AB - To understand how estrogen-responsive genes are regulated, we compared the abilities of estrogen receptors (ERs) alpha and beta to bind to and activate transcription through the consensus vitellogenin A2 ERE and the imperfect pS2, vitellogenin B1, and oxytocin (OT) EREs. Transient transfection experiments demonstrated that ERalpha and ERbeta induced the highest levels of transcription with the A2 ERE, intermediate levels of transcription with the OT ERE, and low levels of transcription with the pS2 and B1 EREs. ERalpha and ERbeta had higher affinities for the A2 ERE than for any of the three imperfect EREs but similar affinities for the pS2, B1, and OT EREs in gel mobility shift assays. ERalpha had a higher affinity and was a more potent activator of transcription than ERbeta. Interestingly, protease sensitivity assays demonstrated that A2, pS2, B1, and OT EREs induced distinct changes in ERalpha and ERbeta conformation thereby providing different functional surfaces for interaction with regulatory proteins involved in control of estrogen-responsive genes. PMID- 11476950 TI - A novel glucocorticoid regulatory unit mediates the hormone responsiveness of the beta1-adrenergic receptor gene. AB - The effects of glucocorticoids on expression of the beta1-adrenergic receptor (beta1AR) gene have been varied. To study the mechanism underling hormonal regulation of the beta1AR, transient transfection of progressively deleted ovine beta1AR promoter fragments was used to identify a 43-bp region (-1274 to -1232 from the translation start site) that contains a novel glucocorticoid regulatory unit (GRU) and confers glucocorticoid responsiveness. Using DNase I footprinting and electrophoretic mobility shift assays (EMSA), we demonstrated the GRU was composed of a palindrome, 5'-TAATTA-3', which is a core binding motif for the homeodomain proteins, an E-box (5'-CACGTG-3'), binding site for the Myc/Max family proteins, and an overlapping glucocorticoid response element (GRE) half site (5'-TGTTCT-3'). EMSA demonstrated that the GRE half-site is critical for GRU protein interactions, which also require binding of proteins to the E-box and the homeodomain region. Co-transfection of a plasmid expressing a c-myc antisense construct significantly reduced glucocorticoid responsiveness of the ovine beta1AR promoter. Furthermore, expression of proteins binding to the GRU was shown to be developmentally regulated, being high in embryonic, reduced in newborn and not detectable in adult heart. We conclude that the ovine beta1AR promoter contains a novel, functional GRU and that glucocorticoid receptor (GR) and the Myc/Max family proteins are involved in the cell-specific nuclear factor binding and transactivation via this element. The results suggest an alternative pathway through which glucocorticoids may exert their effects on genes lacking a full consensus GRE. PMID- 11476951 TI - Swelling-induced changes in cytosolic [Ca2++] in insulin-secreting cells: a role in regulatory volume decrease? AB - Exposure of insulin-secreting cells to hypotonic solutions causes cell swelling followed by regulatory volume decrease (RVD). We have previously demonstrated that RVD is due to activation of a Cl(-) conductance. The present study investigates whether changes in cytosolic [Ca(2+)] play a role in these responses. Hypotonic swelling of RINm5F insulinoma cells caused a marked increase in cytosolic [Ca(2+)]. This effect was abolished by omission of extracellular Ca(2+), by the Ca(2+) channel blockers D600 or Gd(3+)and by 4,4' dithiocyanatostilbene-2,2'-disulphonic acid (DIDS), an inhibitor of the volume sensitive anion. RVD was markedly impaired in the absence of extracellular Ca(2+), but not by D600 nor by Gd(3+). RVD was also inhibited by the maxi-K(+) (BK(Ca)) channel blockers tetraethylammonium (TEA) and iberiotoxin (IbTx), whereas the K(ATP) channel blocker tolbutamide was ineffective. Cell swelling was accompanied by activation of a K(+) conductance which was sensitive to TEA and IbTx but not to tolbutamide. It is concluded that cell swelling causes activation of the volume-sensitive anion channel, leading to depolarization and Ca(2+) entry via voltage-gated Ca(2+) channels. RVD is a Ca(2+)-dependent process, requiring low 'resting' levels of intracellular [Ca(2+)]. However, the swelling-induced increase in cytosolic [Ca(2+)] is not required for RVD to occur. RVD depends upon simultaneous activation of Cl(-) and K(+) channels. We suggest that the BK(Ca) channel is the major K(+) conductance involved in RVD. PMID- 11476952 TI - Alterations of the insulin-like growth factor system in patients with polycythemia vera. AB - The molecular etiology of Polycythemia vera (PV) is still undetermined. Recently, enhanced tyrosine phosphorylation of the insulin-like growth factor-I receptor (IGF-IR) has been shown in PV bone marrow progenitors and peripheral blood mononuclear cells (PBMNC), and elevated levels of IGF binding protein-1 (IGFBP-1) in the serum of PV patients have been reported. To identify further alterations of circulating IGFBPs, the IGFBP profile in the serum of 12 PV patients was compared with age- and sex-matched controls by Western ligand blot (WLB), two dimensional WLB, IGFBP-3 immunoblot and specific RIA for IGFBP-1, -2, -3 and IGFBP-4. To elucidate a role for the IGF-IR in the pathogenesis of PV, basal and IGF-I stimulated tyrosine phosphorylation of the IGF-IR beta-subunit in PBMNC of PV patients or controls was determined by WLB. Furthermore, exons 2, 3 and 15-21 of the IGF-IR were screened for mutations by PCR-single strand conformation polymorphism analysis (PCR-SSCP). We found alterations of the IGFBP profile in the serum of eight out of 12 examined patients including elevated levels of IGFBP 1, -2 and -4, decreased levels of IGFBP-3 and an increase in IGFBP-3 fragment. However, no differences in tyrosine phosphorylation of the IGF-IR in PV patients, neither basal nor IGF-I induced, were detected. Furthermore, no mutations within the screened exons of the IGF-IR could be identified by PCR-SSCP. We conclude that there is no direct impairment of IGF-IR structure or function, but an altered IGFBP profile in a significant portion of PV patients which might contribute to the pathogenesis of PV in these patients. PMID- 11476953 TI - Gene expression of glucocorticoid receptor alpha and beta in giant cell tumour of bone: evidence of glucocorticoid-stimulated osteoclastogenesis by stromal-like tumour cells. AB - Glucocorticoids have been shown to increase bone resorption in vitro and in vivo, however, the mechanism(s) of this action are not fully understood. Given that human giant cell tumour of bone (GCT) is considered to arise from mesenchymal stromal cells and has the capacity to recruit and harbour macrophages and multinucleated osteoclasts, we have used GCT as a model for studying the effect of glucocorticoids on osteoclast formation. We have demonstrated, by RT-PCR and fluorescence in-situ hybridisation, that both glucocorticoid receptor alpha and beta (GRalpha and GRbeta) gene transcripts were present in the stromal-like tumour cells, macrophage-like cells (putative osteoclast precursors) and multinucleated osteoclast-like cells. Moreover, in the presence of 1,25(OH)(2)D(3), dexamethasone dose-dependently stimulated the formation of osteoclast-like cells from GCT-derived co-culture system of stromal-like tumour cells and macrophage-like cells. The stimulation of osteoclastogenesis by dexamethasone was coincident with the up-regulation of receptor activator of NF kappaB ligand (RANKL) but down-regulation of osteoprotegerin (OPG) gene expression in stromal-like tumour cells. These data are consistent with the hypothesis that glucocorticoids increase bone resorption by promoting osteoclastogenesis which is at least in part due to the stimulation of RANKL and inhibition of OPG production in bone stromal cells. PMID- 11476954 TI - Down regulation of melanin concentrating hormone in virally induced obesity. AB - Obesity is a complex disease involving genetic components and environmental factors and probably associated with the dysregulation of central homeostasis normally maintained by the hypothalamic neuroendocrine/neurotransmitter network. We previously reported that canine distemper virus (CDV), which is closely related to human measles virus, can target hypothalamic nuclei, and lead to obesity syndrome in the late stages of infection. Here, using differential display PCR, we demonstrate specific down-regulation of melanin-concentrating hormone precursor mRNA (ppMCH) in infected-obese mice. Although ppMCH was down regulated in all infected mice during the acute stage of infection, this was only seen during the late stage of infection in infected-obese mice. In addition, ppMCH mRNA and protein expression in the lateral hypothalamus was decreased in the absence of neuronal death. These results show the importance of ppMCH in the establishment and maintenance of obesity and the involvement of a virus as an environmental factor. PMID- 11476955 TI - Transforming growth factor-alpha stimulates proliferation of rat Sertoli cells. AB - The number of Sertoli cells is positively correlated with the number of germ cells produced in the testis, but the regulation of Sertoli cell proliferation and final density is poorly understood. Using non-aggregated Sertoli cells from 8 to 9-day-old rat testes, highly enriched by lectin binding, we explored effects of Sertoli cell growth factor candidates in vitro. Proliferation was assessed by 3H-thymidine incorporation, bromodeoxyuridine labeling and supravital staining, and FSH was used as positive control. Transforming growth factor-alpha (TGF alpha) was found to stimulate Sertoli cell proliferation in a dose-dependent manner. Epidermal growth factor (EGF) and betacellulin mimicked the effect, demonstrating specificity of the response as they share receptors with TGF-alpha. Insulin-like growth factor I and II, acidic and basic fibroblast growth factor and stem cell factor lacked significant stimulatory effects. We conclude that EGF/TGF-alpha is a growth factor for Sertoli cells in vitro, possibly contributing to paracrine regulation of Sertoli cell proliferation in vivo. PMID- 11476956 TI - The role of hinge domain in heterodimerization and specific DNA recognition by nuclear receptors. AB - Four structural domains are characteristic of the members of the nuclear receptor superfamily. The hinge (D) domain which is located between the DNA binding (C) domain and the ligand binding (EF) domain, is less conserved among the nuclear receptors. In this study, we investigated the effects of the D domain on receptor function with regard to ligand binding, protein-protein interaction and DNA recognition. We found that EF domain of TR lacked T3 binding activity and additional D domain was required for its ligand binding. Using pull down assays and two-hybrid assays, we also demonstrated that the EF domain of TR did not dimerize with TR or RXR in solution, while the DEF domain was able to homo-and heterodimerize with RXR. In contrast, the RXR EF domain alone was able to heterodimerize with TR. The D domain of TR is required but that of RXR is not necessary for the interaction. We further demonstrated that the D domain was required for receptor specific DNA recognition. The ABC domain of vitamin D receptor (VDR) and TR(DEF) chimeric receptor could not bind to VDR response element (VDRE). Addition of own D domain of VDR to the ABC domain enables the chimeric receptor to bind VDRE and transactivate. The D domain of TR cannot substitute for that of VDR in context of specific DNA recognition. These data suggest that the D domain is important to maintain the integrity of the functional structure of the nuclear receptors. PMID- 11476957 TI - Combined treatment with specific ligands for PPARgamma:RXR nuclear receptor system markedly inhibits the expression of cytochrome P450arom in human granulosa cancer cells. AB - Our previous study demonstrated that PPARgamma specific ligand troglitazone (TGZ) or RXR specific ligand LG100268 (LG) alone decreased the aromatase activity in cultured human ovarian granulosa cells from pre-ovulatory follicles, and combined treatment caused an even greater reduction in this activity. Since similar manners of effects of TGZ or/and LG on the aromatase activity in human ovarian granulosa cancer cell line were observed, we performed the detailed analysis of the mechanisms of these effects using this cell line. The changes in the aromatase activity were associated with comparable changes in the P450arom mRNA levels based on a RNase protection assay. A nuclear run-on assay indicated the P450arom transcript to decrease by 40 and 66% at 24 and 48 h, respectively, after TGZ plus LG treatment. An RNA stability analysis showed the half-life of P450arom mRNA to decrease from 13 to 9 h after the TGZ plus LG treatment. The inhibitory effect of TGZ plus LG on the aromatase activity and P450arom mRNA may not be mediated by the cAMP-PKA pathway that is usually implicated in the regulation of aromatase activity in granulosa cells: because (1) the aromatase activity stimulated by forskolin was not inhibited by TGZ plus LG; (2) the specific PKA inhibitor H89 could not block the inhibitory effect of TGZ plus LG on the aromatase activity; and (3) the luciferase activity of P450arom promoter II did not decrease by the addition of TGZ and LG in transfected cells either at a basic state or in the states stimulated by forskolin or PGE2, respectively. Taken together, these results indicate that TGZ plus LG inhibited the aromatase activity and also decreased the P450arom mRNA level in granulosa cancer cells, and the loss of P450arom mRNA expression was considered to be due to both the decreased transcription and rapid degradation of its RNA. PMID- 11476958 TI - Modulation of rat liver mitochondrial antioxidant defence system by thyroid hormone. AB - In the present study the effect of thyroid hormone (T(3)) on oxidative stress parameters of mitochondria of rat liver is reported. Hypothyroidism is induced in male adult rats by giving 0.05% propylthiouracil (PTU) in drinking water for 30 days and in order to know the effect of thyroid hormone, PTU-treated rats were injected with 20 microg T(3)/100 g body weight/day for 3 days. The results of the present study indicate that administration of T(3) to hypothyroid (PTU-treated) rats resulted in significant augmentation of oxidative stress parameters such as thiobarbituric acid reactive substances and protein carbonyl content of mitochondria in comparison to its control and euthyroid rats. The hydrogen peroxide content of the mitochondria of liver increased in hypothyroid rats and was brought to a normal level by T(3) treatment. Induction of hypothyroidism by PTU treatment to rats also resulted in the augmentation of total and CN-sensitive superoxide dismutase (SOD) activities of the mitochondria, which was reduced when hypothyroid rats were challenged with T(3). Although CN-resistant SOD activity of the mitochondria remained unaltered in response to hypothyroidism induced by PTU treatment, its activity decreased when hypothyroid rats were injected with T(3). The catalase activity of the mitochondria decreased significantly by PTU treatment and was restored to normal when PTU-treated rats were given T(3). Total, Se-independent and Se-dependent glutathione peroxidase activities of the mitochondria were increased following PTU treatment and reduced when T(3) was administered to PTU-treated rats. The reduced and oxidised glutathione contents of the mitochondria of liver increased significantly in hypothyroid rats and their level was restored to normal when hypothyroid rats were injected with T(3). The results of the present study suggest that the mitochondrial antioxidant defence system is considerably influenced by the thyroid states of the body. PMID- 11476959 TI - Similarity of the yellow chromophores isolated from human cataracts with those from ascorbic acid-modified calf lens proteins: evidence for ascorbic acid glycation during cataract formation. AB - Chromatographic evidence supporting the similarity of the yellow chromophores isolated from aged human and brunescent cataract lenses and calf lens proteins ascorbylated in vitro is presented. The water-insoluble fraction from early stage brunescent cataract lenses was solubilized by sonication (WISS) and digested with a battery of proteolytic enzymes under argon to prevent oxidation. Also, calf lens proteins were incubated with ascorbic acid for 4 weeks in air and submitted to the same digestion. The percent hydrolysis of the proteins to amino acids was approximately 90% in every case. The content of yellow chromophores was 90, 130 and 250 A(330) units/g protein for normal human WISS, cataract WISS and ascorbate modified bovine lens proteins respectively. Aliquots equivalent to 2.0 g of digested protein were subjected to size-exclusion chromatography on a Bio-Gel P-2 column. Six peaks were obtained for both preparations and pooled. Side by side thin-layer chromatography (TLC) of each peak showed very similar R(f) values for the long wavelength-absorbing fluorophores. Glycation with [U-(14)C]ascorbic acid, followed by digestion and Bio-Gel P-2 chromatography, showed that the incorporated radioactivity co-eluted with the A(330)-absorbing peaks, and that most of the fluorescent bands were labeled after TLC. Peaks 2 and 3 from the P-2 were further fractionated by preparative Prodigy C-18 reversed-phase high performance liquid chromatography. Two major A(330)-absorbing peaks were seen in peak 2 isolated from human cataract lenses and 5 peaks in fraction 3, all of which eluted at the same retention times as those from ascorbic acid glycated calf lens proteins. HPLC fractionation of P-2 peaks 4, 5 and 6 showed many A(330) absorbing peaks from the cataract WISS, only some of which were identical to the asorbylated proteins. The major fluorophores, however, were present in both preparations. These data provide new evidence to support the hypothesis that the yellow chromophores in brunescent lenses represent advanced glycation endproducts (AGEs) probably due to ascorbic acid glycation in vivo. PMID- 11476960 TI - Cholera toxin-induced PGE(2) activity is reduced by chemical reaction with L histidine. AB - Mediators of cholera toxin (CT)-induced fluid secretion include 3',5'-adenosine monophosphate (cAMP), prostaglandin E(2) (PGE(2)), and 5-hydroxytryptamine (5 HT). Administration of L-histidine significantly reduced the net secretory response of the small intestine of mice challenged with CT and reduced the capacity of PGE(2) to stimulate Na+ transport in Ussing chambers. We demonstrated that L-histidine chemically modified the structure of PGE(2) but had no direct effect on cAMP or 5-HT. L-Histidine and imidazole reacted with PGE(2) in vitro in cell-free mixtures incubated at 37 degrees C and pH 7.0 under an atmosphere of N(2) with the formation of PGE(2)-imidazole and PGE(2)-histidine covalent adducts. Nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) analysis of the purified adduct showed that imidazole catalyzed the dehydration of PGE(2). A Michael adduct then was formed between C11 of 11-deoxy-Delta(10) PGE(2) (PGA(2)) and the tau nitrogen in the imidazole ring of L-histidine. Importantly, the isolated PGE(2)-imidazole and PGE(2)-histidine adducts inhibited CT-induced fluid loss and cAMP accumulation in mouse intestinal loops. The protection provided by PGE(2)-imidazole, PGE(2)-histidine, and L-histidine against intestinal fluid loss could provide a basis for future therapy against cholera. PMID- 11476961 TI - Homogeneous assay based on 52 primer sets to scan for mutations of the ABCA1 gene and its application in genetic analysis of a new patient with familial high density lipoprotein deficiency syndrome. AB - Familial high-density lipoprotein (HDL)-deficiency syndromes are caused by mutations of the ABCA1 gene, coding for the ATP-binding cassette transporter 1. We have developed a homogeneous assay based on 52 primer sets to amplify all 50 ABCA1 exons and approximately 1 kb of its promoter. The assay allows for convenient amplification of the gene from genomic DNA and easy mutational analysis through automatic sequencing. It obviates the need to use mRNA preparations, which were difficult to handle and posed a risk to miss splice junction or promoter mutations. The application of the test to the molecular analysis of a new patient with familial HDL-deficiency (Tangier disease) led to a discovery of two novel ABCA1 mutations: C2665del and C4457T. PMID- 11476962 TI - The role of Kupffer cell alpha(2)-adrenoceptors in norepinephrine-induced TNF alpha production. AB - Although previous studies have demonstrated that plasma levels of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) increase during early sepsis, the precise mechanism responsible for its upregulation remains to be elucidated. Since recent studies have shown that the gut is an important source of norepinephrine (NE) release during early sepsis and enterectomy prior to the onset of sepsis attenuates TNF-alpha production, we hypothesized that gut derived NE plays a major role in upregulating TNF-alpha via the activation of alpha(2)-adrenoceptors on Kupffer cells. To confirm that NE increases TNF-alpha synthesis and release, Kupffer cells were isolated from normal rats and incubated with NE (20 or 50 nM) or another alpha(2)-adrenergic agonist clonidine (50 nM) without addition of Escherichia coli endotoxin. Supernatant levels of TNF-alpha were then measured. In additional animals, intraportal infusion of NE (20 microM) with or without the specific alpha(2)-adrenergic antagonist yohimbine (1 mM) at a rate of 13 microl/min was carried out for 2 h. Plasma and Kupffer cell levels of TNF-alpha were assayed thereafter. Moreover, the effects of NE and yohimbine on TNF-alpha production was further examined using an isolated perfused liver preparation. The results indicate that both NE and clonidine increased TNF-alpha release by approximately 4-7-fold in the isolated cultured Kupffer cells. Similarly, intraportal infusion of NE in vivo or in isolated livers increased TNF alpha synthesis and release which was inhibited by co-infusion of yohimbine. Furthermore, the increased cellular levels of TNF-alpha in Kupffer cells after in vivo administration of NE was also blocked by yohimbine. These results, taken together, suggest that gut-derived NE upregulates TNF-alpha production in Kupffer cells through an alpha(2)-adrenergic pathway, which appears to be responsible at least in part for the increased levels of circulating TNF-alpha observed during early sepsis as well as other pathophysiologic conditions such as trauma, hemorrhagic shock, or gut ischemia/reperfusion. PMID- 11476963 TI - Expression of BRI-amyloid beta peptide fusion proteins: a novel method for specific high-level expression of amyloid beta peptides. AB - In order to develop transgenic animal models that selectively overexpress various Abeta peptides, we have developed a novel expression system that selectively expresses Abeta40 or Abeta42 in the secretory pathway. This system utilizes fusion constructs in which the sequence encoding the 23-amino-acid ABri peptide at the carboxyl terminus of the 266-amino-acid type 2 transmembrane protein BRI is replaced with a sequence encoding either Abeta40 or Abeta42. Constitutive processing of the resultant BRI-Abeta fusion proteins in transfected cells results in high-level expression and secretion of the encoded Abeta peptide. Significantly, expression of Abeta42 from the BRI-Abeta42 construct resulted in no increase in secreted Abeta40, suggesting that the majority of Abeta42 is not trimmed by carboxypeptidase to Abeta40 in the secretory pathway. PMID- 11476964 TI - Investigation of the functional role of active site loop II in a hypoxanthine phosphoribosyltransferase. AB - Hypoxanthine phosphoribosyltransferases (HPRTs) are of biomedical interest because defects in the enzyme from humans can result in gouty arthritis or Lesch Nyhan syndrome, and in parasites these enzymes are potential targets for antiparasite chemotherapy. In HPRTs, a long flexible loop (active site loop II) closes over the active site during the enzyme catalyzed reaction. Functional roles for this loop have been proposed but have yet to be substantiated. For the present study, seven amino acids were deleted from loop II of the HPRT from Trypanosoma cruzi to probe the functional role of this active site loop in catalysis. The mutant enzyme (Deltaloop II) was expressed in bacteria, purified by affinity chromatography, and kinetic constants were determined for substrates of both forward (purine salvage) and reverse (pyrophosphorolysis) reactions catalyzed by the enzyme. Loop II deletion resulted in moderate (0.6-2.7-fold) changes in the Michaelis constants (K(m)s) for substrates other than pyrophosphate (PP(i)), for which there was a 5.8-fold increase. In contrast, k(cat) values were severely affected by loop deletion, with rates that were 240 840-fold below those for the wild-type enzyme. Together with previously reported structural data, these results are consistent with active site loop II participating in transition-state stabilization by precise positioning of the substrates for in line nucleophilic attack and in the liberation of PP(i) as a product of the salvage reaction. PMID- 11476965 TI - Novel mutations in ABCA1 gene in Japanese patients with Tangier disease and familial high density lipoprotein deficiency with coronary heart disease. AB - Mutations in the ATP-binding cassette transporter 1 (ABCA1) gene have been recently identified as the molecular defect in Tangier disease (TD) and familial high density lipoprotein deficiency (FHA). We here report novel mutations in the ABCA1 gene in two sisters from a Japanese family with TD who have been described previously (S. Ohtaki, H. Nakagawa, N. Kida, H. Nakamura, K. Tsuda, S. Yokoyama, T. Yamamura, S. Tajima, A. Yamamoto, Atherosclerosis 49 (1983)) and a family with FHA. Both probands of TD and FHA developed coronary heart disease. Sequence analysis of the ABCA1 gene from the patients with TD revealed a homozygous G to A transition at nucleotide 3805 of the cDNA resulting in the substitution of Asp 1229 with Asn in exon 27, and a C to T at nucleotide 6181 resulting in the substitution of Arg 2021 with Trp in exon 47. Sequence analysis of the ABCA1 gene from the FHA patient revealed a homozygous 4 bp CGCC deletion from nucleotide 3787 to 3790 resulting in premature termination by frameshift at codon 1224. These mutations were confirmed by restriction digestion analysis, and were not found in 141 control subjects. Our findings indicate that mutations in the ABCA1 gene are associated with TD as well as FHA. PMID- 11476966 TI - Prolonged Jun N-terminal kinase (JNK) activation and the upregulation of p53 and p21(WAF1/CIP1) preceded apoptosis in hepatocytes after partial hepatectomy and cisplatin. AB - Cisplatin induced apoptosis in regenerating liver after partial hepatectomy (PH). Apoptosis was determined by in situ end-labeling and gel electrophoresis of DNA fragmentation. Characteristic DNA fragmentation was obvious at 4 h and peaked at 8 h after PH. The activity of Jun N-terminal kinase (JNK) transiently increased at 1 h after PH. However, in cisplatin-injected rats, the JNK activity increased at 30 min and the increased level was maintained up to 4 h after PH. The in vivo activation of JNK was confirmed by the increased level of the phosphorylated c Jun protein. Western blot analysis showed that the phosphorylated c-Jun level increased at 1 h and reached more than 30-fold the control level at 2 h after PH with cisplatin. The c-jun mRNA levels also markedly increased at 1 h after PH with cisplatin. The protein level of p53 increased after 1 h on cisplatin injection, but no significant change in the mRNA level was observed. The rise in the p53 protein level was followed by the upregulation of p21(WAF1/CIP1) mRNA and protein levels. These results suggested that the enhanced and sustained JNK activation and the upregulation of p53 and p21(WAF1/CIP1) were involved in hepatocyte apoptosis induced by PH with cisplatin. PMID- 11476967 TI - Age-related responses of the microcirculation to ischemia-reperfusion and inflammation. AB - Aging is a major risk factor for a variety of ischemic disorders including ischemic heart disease and stroke. Intense research over the past decade into ischemia-reperfusion (I/R) injury has implicated a general mechanism whereby reactive oxygen species produced at the onset of reperfusion overwhelm endogenous antioxidants, resulting in a cascade of events including mast cell degranulation, recruitment of neutrophils to the endothelial wall, arteriolar constriction that limits tissue perfusion, and increased vascular permeability that leads to inflammation and edema. Much of our knowledge regarding I/R injury comes from animal models; however, despite the fact that I/R disproportionately affects older individuals, young animals are usually chosen in models of I/R injury due to their greater availability, lower cost, and fewer health problems. Results obtained from young animals demonstrate a central role for both neutrophils and mast cells in I/R-induced increases in microvascular permeability and arteriolar constriction; however, it is not clear that a role for neutrophils is extended to older animals. A growing body of evidence indicates that neutrophils isolated from elderly individuals exhibit attenuated chemotaxis, oxidant release, and phagocytosis, and it has been suggested that these deficiencies are related to an age-associated increase in glucocorticoid production and oxidative stress. Therefore, neutrophils may have a limited capacity to influence microcirculatory tissue in the elderly compared to in the young. In support of this hypothesis, I/R-induced increases in microvascular permeability and decreases in vascular perfusion have been found to occur in older rats despite the absence of a significant increase in leukocyte-endothelial cell adhesion. Furthermore, elimination of circulating neutrophils attenuates I/R-induced mesenteric permeability only in young rats. Therefore, it appears that neutrophil independent mechanisms of inflammation may be responsible for much of the microvascular dysfunction initiated by I/R in older animals. PMID- 11476968 TI - Liver dysfunction elicited by gut ischemia-reperfusion. AB - Gut ischemia and reperfusion (I/R) has been implicated as a prime mechanism in the pathogenesis of multiple organ failure and in initiating remote organ failure. Although it has long been known that gut I/R elicits liver dysfunction, only recently has the kinetics of leukocyte accumulation in the hepatic microcirculation and mechanisms of the liver injury after gut I/R been investigated. These studies reveal that the magnitude of gut I/R-induced liver injury depends on the duration of ischemic period and animal species. Gut I/R induced accumulation of leukocytes, both neutrophils and lymphocytes, in the liver results in an oxidative stress in proximity to non-perfused sinusoid that contributes to subsequent hepatocellular injury. The gut I/R-induced leukosequestration in the liver is mediated by adhesion molecules that are induced by different cytokines, endotoxin, and oxidants. Kupffer cells also play an important role in the gut I/R-induced leukosequestration and liver injury. Nitric oxide and anti-oxidants such as superoxide dismutase protect the liver against the deleterious effects of gut I/R. Furthermore, agents such as ethanol can alter the hepatic responses to gut I/R. The results of these studies provide novel information and potential therapeutic strategies for reducing the liver dysfunction and multiple organ failure induced by gut I/R. PMID- 11476969 TI - Reduced susceptibility of low density lipoprotein to lipid peroxidation after cholestyramine treatment in heterozygous familial hypercholesterolemic children. AB - Cholestyramine treatment in children with heterozygous familial hypercholesterolemia (FHHe) can interfere with fat absorption from the intestinal tract, and has the potential to decrease the absorption of fat-soluble vitamins. The aim of this study was to examine the effect of cholestyramine treatment on the levels of the fat soluble vitamins (vitamin E, beta-carotene and lycopene) in LDL, on the glutathione system and on the susceptibility of LDL to oxidation in FHHe children. Patients were 16 children (seven girls, nine boys), age 14+/-4 years, non-smokers. Plasma LDL level before cholestyramine treatment but after dietary treatment was 239+/-50 mg% with no secondary cause for hypercholesterolemia. A control group was comprised of ten children (seven girls, three boys), age 14+/-4 years with plasma LDL level of 100+/-14 mg%. Blood was drawn from 16 FHHe children and five control children after fasting for 14 h. Thereafter cholestyramine treatment was begun in the patient group, at a dose of 8 g/day for 2 months. At the end of this period the dose was increased to 12-16 g/day for an additional 2 months. After 4 months from the beginning of the treatment, blood was drawn again. Plasma LDL cholesterol decreased after treatment by 14% (from 239+/-67 mg% before treatment to 205+/-55 mg% after treatment, P=0.07). Malondialdehyde (MDA) levels measured by thiobarbituric reactive substances (TBARS) assay in LDL at the end of oxidation were 30% higher in FHHe children in comparison to controls (P=0.02). After treatment TBARS levels in LDL (after in vitro oxidation) from FHHe children were decreased by 23% (P=0.02). Vitamin E levels in LDL from FHHe children after treatment were decreased by 65%, while beta-carotene and lycopene contents in LDL, paradoxically increased by 90 and 102%, respectively. In red blood cells (RBC), glutathione peroxidase (GPx) and glutathione transferase (GTf) activities were decreased by 29 and 24%, respectively, while glutathione reductase activity, total and oxidized glutathione contents from FHHe children did not change after cholestyramine treatment. LDL was more prone to oxidation in FHHe children than in controls, when measured by TBARS levels after LDL oxidation (with 10 &mgr;M CuSO(4)). Cholestyramine treatment for 4 months normalized LDL susceptibility to oxidation measured by TBARS levels, despite the decrease in vitamin E content in LDL from treated FHHe children. This is presumably due to the increased LDL content of beta-carotene and lycopene after treatment. GPx and GTf activities decreased after treatment, presumably due to the drop in oxidative stress within the RBCs, in parallel to the decreased LDL tendency to oxidation. Cholestyramine treatment in FHHe children has an overall antioxidant effect on LDL. PMID- 11476970 TI - Combined use of adenosine and amrinone inhibits reperfusion injury of rat liver. AB - Although intraportal infusion of adenosine suppressed the oxidative stress caused by activated neutrophils and attenuated ischemia-reperfusion injury of canine liver, high doses of adenosine elicit systemic hypotension. The present work demonstrates that combined use of low doses of adenosine and amrinone, a phosphodiesterase inhibitor, strongly inhibited reperfusion injury of the liver without eliciting hypotension. After 45 min ischemia followed by 60 min reperfusion of rat liver, low doses of adenosine and amrinone were administrated intraportally, resulting in significantly increased hepatic levels of cGMP, cAMP, nitrite plus nitrate in plasma, and decreased alanine aminotransferase in plasma without changing hemodynamics. Thus, intraportal administration of low doses of adenosine and amrinone increased the cyclic nucleotides, thereby improved microcirculation and attenuated reperfusion injury of the liver. PMID- 11476971 TI - Laser irradiation as a potential pathogenetic method for immunocorrection in rheumatoid arthritis. AB - We investigated the immunocorrective and clinical effect in 75 rheumatoid patients treated with intravenous laser blood irradiation. A relation between the positive immunotropic (as well as therapeutic) effect and the pre-existent immune status of each patient was revealed. A well-defined effect was found in patients with a low level of CIC and a normal count of functional-competent T-cells. ILIB provided some symptomatic but unstable relief in patients with a high level of CIC and a high functional activity of T-lymphocytes. There was no effect in patients with a high level of CIC and decreased number of lymphocytes. PMID- 11476972 TI - Organ-specific characteristics of blood supply in acute hemorrhage in rats with different resistance to circulatory hypoxia. AB - The present study is aimed toward a comparative real-time analysis of organ specific characteristics of the liver, kidney and brain blood supply in the dynamics of acute massive hemorrhage in rats with different resistance to circulatory hypoxia. The survival time of experimental animals after the arrest of bleeding was used as a criterion of their resistance to acute blood loss. The rats with high resistance (HR) to hypoxia had the survival time not less than 3 h, while the rats with low resistance (LR) to hypoxia lived not more than 1.5 h. A marked decrease in arterial organ blood flow velocity and tissue perfusion of the liver, kidney and brain in LR and HR rats was observed at the end of acute massive hemorrhage in ultrasonic and Doppler flowmetry. In the post-hemorrhagic period the organ hemodynamics and microcirculation showed a tendency to a further decrease in LR rats. In HR animals the blood flow velocities in hepatic, renal and common carotid arteries were temporarily restored to 115-120, 85-90 and 60 65%, respectively, following the bleeding arrest. In the compensated phase of the post-hemorrhagic period the brain blood flow was maintained at this new post hemorrhagic level due to autoregulatory changes in the carotid resistance. Such a response of brain blood vessels of HR rats is considered to be an adaptive response which protects the brain from autoreperfusion- and reoxygenation-induced injuries under conditions of posthemorrhagic autorestoration of tissue circulation. PMID- 11476973 TI - The spatial patterns of pathological brain lesions in 12 patients with corticobasal degeneration. AB - Corticobasal degeneration (CBD) is a rare and progressive neurological disorder characterised by the presence of ballooned neurons (BN) and tau positive inclusions in neurons and glial cells. We studied the spatial patterns of the BN, tau positive neurons with inclusions (tau+ neurons), and tau positive plaques in the neocortex and hippocampus in 12 cases of CBD. All lesions were aggregated into clusters and in many brain areas, the clusters were distributed in a regular pattern parallel to the tissue boundary. In the majority of cortical areas, the clusters of BN were larger in the lower compared with the upper laminae while the clusters of tau+ neurons were larger in the upper laminae. Clusters of BN and tau+ neurons were either negatively correlated or not significantly correlated in the upper and lower cortical laminae. Hence, BN and tau+ lesions in CBD exhibit similar spatial patterns as lesions in Alzheimer's disease (AD), dementia with Lewy bodies (DLB) and Pick's disease (PD). The location, sizes and distribution of the clusters in the neocortex suggest that the tau+ lesions may be associated with the degeneration of the feedforward and the BN the feedback cortico-cortical and/or the efferent cortical pathways. PMID- 11476974 TI - Diaphragmatic function in patients with chronic left ventricular failure. AB - The aim of this paper is to evaluate the function of respiratory muscles in patients with chronic moderate left ventricular failure (LVF), and its contribution to the pathophysiology of dyspnea and fatigue. We have studied 20 male patients with LVF, classes II and III of New York Heart Association (NYHA), mean age 66.9+/-10 years (GI) and 19 male aged-matched controls without cardiopulmonary disease mean age 64.6+/-8.4 years (GII). The evaluation included (a) methods derived from volitional manoeuvres, maximal inspiratory pressures at Functional Residual Capacity; maximal expiratory pressures at total lung capacity (TLC); nasal sniff; oesophageal sniff and transdiaphragmatic pressures; (b) methods derived from non-volitional manoeuvres, using bilateral cervical magnetic stimulation of the phrenic nerves, measuring the following twitch pressures (oesophageal, gastric and the transdiaphragmatic). With volitional manoeuvres we have not found statistically significant differences between the two groups: maximal expiratory pressures (cmH(2)O), GI 138+/-42; GII 152+/-40; P=NS and maximal inspiratory pressures (cmH(2)O), GI 74.1+/-22; GII 85+/-16; P=NS. However, these values were significantly lower than those obtained with sniff manoeuvres, nasal sniff (cmH(2)O), GI 95.6+/-22; GII 99.6+/-16; P=NS and oesophageal sniff (cmH(2)O), GI 96.2+/-20; GII 97.5+/-18; P=NS. There were no significant differences between nasal sniff and oesophageal sniff. Using cervical magnetic stimulation, we also didn't find a significant difference for transdiaphragmatic twitch between groups, but the contribution of the diaphragm to the transdiaphragmatic pressure was lower in patients with LVF since the oesophageal twitch was lower (cmH(2)O), GI 11.4+/-3.4; GII 16.3+/-6.8; P<0,004. In conclusion, the contribution of the diaphragm to total ventilation in-patients with moderate LVF is preserved. However, its capacity to generate negative intra thoracic pressures is decreased since there is a significant decrease in oesophageal twitch. So, it seems that the diaphragm is the first inspiratory muscle to be affected in patients with moderate LVF. PMID- 11476975 TI - Association of immunological disorders in lethal side effect of NSAIDs on beta glucan-administered mice. AB - (1-->3)-beta-D-Glucan (beta-glucan) is a biological response modifier that regulates host immune response. We have found that the combination of a beta glucan and a non-steroidal anti-inflammatory drug (NSAID), indomethacin (IND), induced lethal toxicity in mice [Yoshioka et al. (1998) FEMS Immunol. Med. Microbiol., 21, 171-179]. This study was undertaken to analyze the mechanism of the lethal side effect. Combination of a beta-glucan and IND increased the number of leukocytes, especially macrophages and neutrophils, in various organs and these cells were activated. The activated state of these cells was supported by the enhanced production of interferon-gamma in the presence of IND in vitro culture of the peritoneal exudate cells. Intestinal bacterial flora was translocated into the peritoneal cavity in these mice to cause peritonitis. Comparing the toxicity of various NSAIDs, nabumetone, a partially cyclooxygenase 2-selective NSAID with weaker toxicity to the gastrointestinal tract, did not exhibit a lethal side effect. These facts strongly suggested that gastrointestinal damage by NSAIDs was more severe in beta-glucan-administered mice, resulting in peritonitis by enteric bacteria and leading to death. PMID- 11476976 TI - New restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus. AB - In this study, we isolated and tested restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus based on PCR products amplified by the random amplified polymorphic DNA (RAPD) primer R108. Four DNA fragments, Afd, Af5, Af4, and Af4A, were amplified. Fragments Afd and Af5 were 85% and 88% identical at the DNA level to part of the Afut1 retrotransposon from A. fumigatus. Fragment Af4A is a duplication of fragment Af4 and both showed similarity at the amino acid level with endonucleases from other fungal retrotransposons. We used both RAPD with primer R108 and RFLP assays with Afut1, Afd, and Af4A, to determine the genetic relatedness of clinical isolates of A. fumigatus isolated sequentially from four patients colonized with A. fumigatus. The combination of these different methods suggested that the isolates infecting the four patients were not identical. PMID- 11476977 TI - Serum antibodies to group A streptococcal extracellular and cell-associated antigens in Egyptians with post-streptococcal diseases. AB - We investigated serum antibodies to a comprehensive array of group A streptococcal antigens and superantigens in Egyptian subjects. Antibodies to Streptococcus pyogenes cell-associated proteins and to proteins released by rapidly dividing S. pyogenes were compared in four patient groups with different post-streptococcal diseases and in healthy controls. Enzyme-linked immunosorbent assays showed that total Ig and IgG to extracellular antigens were significantly higher in patients with acute rheumatic fever (ARF) compared to healthy controls, but no differences were found in either total Ig or IgG titres to cell-associated proteins between any of the groups. Western blotting showed that multiple extracellular and cell-associated antigens, covering a wide range of molecular masses, were recognised by all sera, including healthy controls. No evidence was obtained for putative dominant antigens associated with any disease group, although a low molecular mass cell-associated protein (approximately 4 kDa) was clearly recognised by two-thirds of subjects irrespective of disease status. These findings demonstrate that raised serum Ig and IgG titres to extracellular, but not cell-associated, S. pyogenes antigens are a feature of ARF in this population, and suggest that multiple S. pyogenes antigens contribute to this response. PMID- 11476978 TI - Flavonoids protect mice from two types of lethal shock induced by endotoxin. AB - The protective effect of flavonoids on two types of lethal endotoxic shock was studied. A lethal endotoxic shock was induced by administration of lipopolysaccharide (LPS) into D-galactosamine (D-GalN)-sensitized mice and another one was done by administration of a high dose of LPS into normal mice. Pretreatment with a series of flavonoids protected mice from two types of endotoxin lethality. Flavonoid pretreatment reduced the serum tumor necrosis factor-alpha (TNF-alpha) level in mice injected with D-GalN and LPS, but not in mice injected with a high dose of LPS. TNF-alpha-induced lethal shock in D-GalN sensitized mice was also protected by pretreatment with flavonoids, suggesting that flavonoids augmented the resistance to TNF-alpha lethality. On the other hand, flavonoids reduced the plasma level of lipid peroxides in mice injected with a high dose of LPS, but not in D-GalN-sensitized mice. Taken together, these results indicated that flavonoids might protect mice from two types of endotoxin lethality. The protective mechanism of flavonoids in each endotoxin lethality is discussed. PMID- 11476979 TI - Good adhesion properties of probiotics: a potential risk for bacteremia? AB - The ability to adhere to human intestinal mucus was tested for lactic acid bacteria of clinical blood culture, human fecal and dairy origin. The blood culture isolates were found to adhere better than the dairy strains. Of the Lactobacillus rhamnosus strains (nine clinical, 10 fecal and three dairy), blood culture isolates adhered better than the fecal strains. Although these results indicate a trend for blood culture isolates to bind to intestinal mucus in higher numbers than strains of dairy and human fecal origin, other factors are also likely to be involved in the etiology of lactobacillemia since some of the clinical Lactobacillus isolates exhibited a relatively low level of adhesion. PMID- 11476980 TI - Importance of the host genetic background on immune responses to Helicobacter pylori infection and therapeutic vaccine efficacy. AB - In order to investigate the role of host factors in Helicobacter pylori infection and immunity, two different strains of inbred mice, C57BL/6 and BALB/c, were infected with a standard H. pylori strain, SS1. A month later, infected mice were immunized orally with whole-cell lysates of H. pylori SS1 and cholera toxin on days 1, 3, 6, 30, and 54. Ten days after the last immunization, mice were sacrificed and the stomach was collected to assess H. pylori colonization density by quantitative culture. H. pylori SS1 colonization was significantly greater in C57BL/6 than in BALB/c (P<0.02 and P<0.003 at 2 and 13 weeks post-inoculation, respectively). Colonization in C57BL/6 persisted at equivalent levels for 13 weeks but the colonization density in BALB/c decreased significantly during this period. In contrast to the pattern of bacterial colonization, antibody responses following H. pylori SS1 infection were greater in BALB/c than in C57BL/6, suggesting that host factors may modulate the immune responses to H. pylori infection. Following therapeutic immunization, H. pylori colonization in BALB/c mice was also significantly reduced (P<0.03), while no significant differences in bacterial density were observed in C57BL/6. These observations collectively demonstrate the great importance of host factors in H. pylori infection and the development of effective immune responses. PMID- 11476981 TI - Cervical secretory immunoglobulin A to human papillomavirus type 16 (HPV16) from HPV16-infected women inhibit HPV16 virus-like particles-induced hemagglutination of mouse red blood cells. AB - Secretory immunoglobulin A (sIgA) antibodies are the first line of defence at the genital mucosa, and are thought to hinder viral infections by binding to conformational epitopes on the viral capsid. To investigate if cervical sIgA binds to conformational epitopes of the Human papillomavirus type 16 (HPV16), cervical mucus samples from 109 HPV16-infected patients were examined in a HPV16 virus-like particles-induced hemagglutination inhibition assay. 48 (44.1%) patients were able to inhibit hemagglutination. Inhibition of hemagglutination was associated with the presence of sIgA (P=0.001). In conclusion, naturally occurring cervical anti-HPV16 sIgA binds to and hinders conformational epitopes on the viral capsid, suggesting that these antibodies might have a neutralizing capacity. PMID- 11476982 TI - Detection of Toll-like receptor 2 (TLR2) mutation in the lepromatous leprosy patients. AB - Toll-like receptor 2 (TLR2) is critical in the immune response to mycobacterial infections and the mutations in the TLR2 have been shown to confer the susceptibility to severe infection with mycobacteria. To define this, we screened the intracellular domain of TLR2 in 131 subjects. Groups of 45 lepromatous and 41 tuberculoid leprosy (TT) patients and 45 controls were investigated. Ten subjects among the lepromatous leprosy (LL) patients had a band variant detected by single stranded conformational polymorphism. DNA sequencing detected a C to T substitution at nucleotide 2029 from the start codon of the TLR2. The mutation would substitute Arg to Trp at amino acid residue 677, one of the conserved regions of TLR2. In our results, the mutation was involved in only LL, not TT and control. Thus, we suggest that the mutation in the intracellular domain of TLR2 has a role in susceptibility to LL. PMID- 11476983 TI - Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting. AB - Tuberculous meningitis (TBM) is one of the commonest chronic infections of the central nervous system (CNS). Diagnosis of TBM has been a problem as it causes various clinical manifestations which can be confused with those of other chronic infections of the CNS such as neurocysticercosis (NCC), neurobrucellosis and cryptococcal meningitis, that are prevalent in many underdeveloped and developing countries. Differential diagnosis of TBM can be made by detecting circulating mycobacterial antigens in CSF by immunoassays. In this study, a reverse passive hemagglutination (RPHA) has been developed using rabbit antimycobacterial IgG for detection of circulating mycobacterial antigens in CSFs from chronic infections of the CNS in order to develop a rapid, simple, sensitive and cost-effective method. Circulating mycobacterial antigens were characterized by immunoblot assay. The sensitivity limit of RPHA was 400 ng ml(-1). RPHA was specific as antimycobacterial IgG did not show any reaction with porcine Cysticercus cellulosae which was used as a control antigen. RPHA could detect mycobacterial antigens in CSF at a sensitivity level of 94.11% with a specificity of 99.0%. Immunoblot analysis of RPHA positive CSFs revealed predominantly 30-32 kDa and 71 kDa antigens whilst 6, 86, 120, 96 and 110 kDa showed varied degree of reactivity. Antigens of masses 30-32 and 71 kDa were absent in culture filtrate of Mycobacterium tuberculosis H37Rv grown in Proskeur-Beck liquid medium. RPHA is a rapid, simple and sensitive immunological method with a long shelf life of 6-8 weeks if stabilized coated erythrocytes are stored at +4 degrees C. RPHA could be used as an additional immunodiagnostic tool in both differential diagnosis and prognosis of TBM. Immunoblot results indicate that 30-32 kDa and 71 kDa antigens are cell wall derived. PMID- 11476984 TI - HIV protease inhibitors attenuate adherence of Candida albicans to epithelial cells in vitro. AB - Oropharyngeal candidiasis is one of the first and most commonly reported opportunistic infections of untreated AIDS patients. With the introduction of the new antiviral HAART therapy, including HIV protease inhibitors, this mucocutaneous infection is nowadays only rarely observed in treated patients. It was recently shown that HIV protease inhibitors have a direct attenuating effect on Candida albicans secreted aspartic proteinases (Saps), an investigation prompted by the fact that both Sap and HIV protease belong to the superfamily of aspartic proteinases and by the observation that mucocutaneous infections sometimes resolve even in the absence of an immunological improvement of the host. As these Saps are important fungal virulence factors and play a key role in adhesion to human epithelial cells we tried to assess the effect of the HIV protease inhibitors Ritonavir, Indinavir and Saquinavir on fungal adhesion to these cells. The effect on phagocytosis by polymorphonuclear leukocytes was also assessed. Ritonavir was found to be the most potent inhibitor of fungal adhesion. A dose-dependent inhibition of adhesion to epithelial cells was found already at 0.8 microM and was significant at 4 microM or higher, at 500 microM the inhibition was about 55%. Indinavir and Saquinavir inhibited significantly at 4 microM or 20 microM, respectively; at 500 microM the inhibition was 30% or 50%. In contrast, no protease inhibitor was able to modulate phagocytosis of Candida by polymorphonuclear leukocytes. In conclusion, inhibition of Saps by HIV protease inhibitors may directly help to ease the resolution of mucosal candidiasis. In future, derivatives of HIV protease inhibitors, being more specific for the fungal Saps, may form an alternative in the treatment of mucosal candidiasis insensitive to currently available antimycotics. PMID- 11476985 TI - Genetic differences among the LPS biosynthetic loci of serovars of Leptospira interrogans and Leptospira borgpetersenii. AB - The gene organization in the lipopolysaccharide biosynthetic (rfb) locus was analyzed in seven Leptospira interrogans serovars within serogroup Icterohemorrhagiae, seven non-Icterohemorrhagiae serovars and one Leptospira borgpetersenii serovar. Two groups of loci were delineated based on DNA hybridization and sequence analysis. Group 1 contained the two Hardjo subtypes, Hardjoprajitno and Hardjobovis. Group 2 (containing Copenhageni, Pomona, Naam, Mwogolo, Smithi, Lai, Canicola, Autumnalis, Pyrogenes, Australis and Icterohemorrhagiae) differed from Group 1 in its organization upstream of orf11, where five ORFs (32, 33, 34, 35, 37) were identified that were not contained in the Group 1 loci. These ORFs encoded a putative epimerase (orf32), a glycosyltransferase (orf33), two integral membrane proteins (orfs 34 and 35), and a galactosyltransferase (orf37). Serovars Australis, Pomona and Autumnalis did not contain orf37. Serovar Bataviae was excluded from the grouping because of its unique genetic organization upstream of orf13. In the Group 2 loci, comparison of the genetic layout at the 5' end revealed differences which included mutations disrupting reading frames in either or both orf34 and orf35 and apparent allelic differences between orf33 homologs that may be sufficient to account for the genetic basis of serovar identity. PMID- 11476987 TI - E-cadherin complex and its abnormalities in human breast cancer. AB - E-cadherin and its adhesion complex play an essential role in the adhesion of breast cancer cells and tissues. Members of the complex, such as beta-catenin, act as regulators on cell adhesion, and are involved in cell signalling and transcription regulation. The adhesion complex is a known structure in the control of tumour progression and metastasis. Recent years have seen a rapid expansion in the understanding of the biology and clinical relevance of the complex in breast cancer. The current article summarises recent progresses in the molecular/cellular biology of E-cadherin and its complex and the clinical, diagnostic, prognostic, and therapeutic value of this complex in breast cancer. PMID- 11476988 TI - Recent advances in carcinoid pathogenesis, diagnosis and management. AB - Carcinoid tumors usually present as diagnostic dilemmas due to obscure or nonspecific symptomatology. Advances in molecular biology are allowing the investigation of molecular markers of aggressiveness, better serum tumor markers, as well as the molecular pathogenesis of carcinoid heart disease. Somatostatin receptor scintigraphy (SRS) and whole body positron emission tomography (PET) are providing much improved sensitivity in localization of both primary and metastatic tumors. Long acting depot somatostatin analogues are combining effectiveness and ease of use for medical management of carcinoid syndrome. An additional benefit may be tumor growth suppression. PMID- 11476989 TI - Gene therapy for pancreatic cancer--current and prospective strategies. AB - Pancreatic ductal adenocarcinoma is one of the most common causes of cancer death in the developed world. Long-term survival is currently only achieved through surgical resection. Most patients have locally advanced or metastatic disease at the time of diagnosis and are therefore not amenable to resection, whilst chemotherapy and radiotherapy are by and large ineffective. Gene therapy offers an alternative to current adjuvant strategies. With approximately two-thirds of all gene therapy trials worldwide directed at cancer, the gene therapy approaches that are currently being explored for pancreatic cancer are specifically examined. Gene delivery systems, genetic targets, and combined gene delivery with chemotherapy are discussed in the context of pancreatic cancer treatment. PMID- 11476990 TI - Management of locally advanced rectal cancer. AB - The treatment of rectal cancer typically involves a multidisciplinary approach. A minority of patients will have tumors that are full thickness, involve adjacent structures, or have metastatic disease to regional lymph nodes. The combination of adjuvant therapy and surgical resection is the mainstay of treatment for locally advanced carcinoma of the rectum. This article will review the role of adjuvant chemotherapy and radiotherapy in patients with high risk tumors. The operative considerations in advanced rectal cancers will be reviewed. In particular, the role of mesorectal excision and exenterative surgery will be discussed. PMID- 11476991 TI - Beware of frontal lobe deficits in hippocampal clothing. AB - The Wisconsin card-sorting test (WCST) is a commonly used clinical tool for the detection of frontal lobe dysfunction, specifically executive dysfunction. Patients with lesions outside the frontal lobes sometimes show deficits on the WCST, however, and some researchers have implicated hippocampal dysfunction as the cause of the deficit. But a critical role for the hippocampus seems to be untenable because amnesic patients with bilateral medial temporal lobe (MTL) lesions perform the WCST normally. In the case of epileptic patients, an alternative explanation of the card-sorting impairment is the propagation of abnormal discharges from MTL to frontal lobe structures, causing remote interference with executive circuits. PMID- 11476993 TI - Stimulating times for TMS. PMID- 11476992 TI - Moving faces. PMID- 11476994 TI - Through the looking glass. PMID- 11476995 TI - Superior temporal gyrus: a neglected cortical area? PMID- 11476996 TI - Rumelhart Prize winner announced. PMID- 11476997 TI - We've all got rhythm. PMID- 11476998 TI - Neglecting the temporal lobe. PMID- 11476999 TI - Tactile motion activates V5. PMID- 11477000 TI - Bugs in vision. PMID- 11477001 TI - Bugs in odometry. PMID- 11477002 TI - The in vivo/in vitro approach to cognition: the case of analogy. AB - A new way of investigating cognition is proposed that combines naturalistic (in vivo) and experimental (in vitro) methods into a single approach. Investigating analogical reasoning by scientists 'live' at laboratory meetings (in vivo) we found that the types of analogies they use change with their goals. Unlike subjects in many cognitive 'reminding' experiments, scientists frequently used structural similarities in their analogies, as opposed to superficial similarities. By subsequently using the in vitro approach, we found that the use of structural information was associated with being asked to generate analogies and with the use of complex scenarios. Similar results are obtained in other areas of reasoning such as hypothesis testing and causal reasoning. PMID- 11477003 TI - On the role of space and time in auditory processing. AB - Unlike visual and tactile stimuli, auditory signals that allow perception of timbre, pitch and localization are temporal. To process these, the auditory nervous system must either possess specialized neural machinery for analyzing temporal input, or transform the initial responses into patterns that are spatially distributed across its sensory epithelium. The former hypothesis, which postulates the existence of structures that facilitate temporal processing, is most popular. However, I argue that the cochlea transforms sound into spatiotemporal response patterns on the auditory nerve and central auditory stages; and that a unified computational framework exists for central auditory, visual and other sensory processing. Specifically, I explain how four fundamental concepts in visual processing play analogous roles in auditory processing. PMID- 11477004 TI - The probabilistic approach to human reasoning. AB - A recent development in the cognitive science of reasoning has been the emergence of a probabilistic approach to the behaviour observed on ostensibly logical tasks. According to this approach the errors and biases documented on these tasks occur because people import their everyday uncertain reasoning strategies into the laboratory. Consequently participants' apparently irrational behaviour is the result of comparing it with an inappropriate logical standard. In this article, we contrast the probabilistic approach with other approaches to explaining rationality, and then show how it has been applied to three main areas of logical reasoning: conditional inference, Wason's selection task and syllogistic reasoning. PMID- 11477005 TI - Shedding light on brain function: the event-related optical signal. AB - One of the basic goals of cognitive psychology is the analysis of the covert processes that occur between stimulus and response. In the past 20-30 years, the tools available to cognitive psychologists have been augmented by a number of imaging techniques for studying the 'brain in action' in a non-invasive manner. These techniques have their strength in either temporal or spatial information, but not both. We review here recent advances of a new approach, the event-related optical signal (EROS). This method allows measurements of the time course of neural activity in specific cortical structures, thus combining good spatial and temporal specificity. As an example, we show how EROS can be used to distinguish between serial and parallel models of information processing. PMID- 11477006 TI - Calculating creative connections. PMID- 11477007 TI - Francisco J. Varela (1946-2001). PMID- 11477008 TI - The "piriformis syndrome"--myth or reality? PMID- 11477011 TI - The use of local anaesthetic injections in professional football. PMID- 11477012 TI - Exercise and outdoor ambient air pollution. AB - OBJECTIVES: To establish by literature survey: (a) levels at which air pollutants are considered damaging to human health and to exercisers in particular; (b) the current ambient levels experienced in the United Kingdom; (c) whether athletes are especially at risk. METHODS: Six major urban air pollutants were examined: carbon monoxide (CO); nitrogen oxides (NO(X)); ozone (O(3)); particulate matter (PM(10)); sulphur dioxide (SO(2)); volatile organic compounds (VOCs). RESULTS: CO is detrimental to athletic performance. NO(2) is of concern to human health, but outdoor levels are low. O(3) poses a potentially serious risk to exercising athletes. Decrements in lung function result from exposure, and there is evidence that athletic performance may be affected. Detrimental effects may occur at low ambient levels, but there is no scientific consensus on this matter. PM(10) is causing concern in the scientific community. Blood lead accumulation during exercise indicates that personal exposure to toxic compounds associated with PM(10) may be magnified. Generally, outdoor ambient levels of SO(2) are too low to cause a problem to the athlete, except the asthmatic athlete. The few studies on exposure of exercisers to VOCs are reviewed. CONCLUSIONS: Athletes and exercisers should avoid exercising by the road side even though levels of the more noxious air pollutants have been controlled in the United Kingdom. O(3) is particularly damaging to athletes; it reaches its highest concentrations on hot bright days in rural areas. PMID- 11477013 TI - Sepsis and mechanisms of inflammatory response: is exercise a good model? AB - OBJECTIVES: The immune changes induced by a bout of prolonged and vigorous exercise have been suggested to be a useful experimental model of sepsis and the inflammatory response. Available literature was reviewed to evaluate this hypothesis. METHODS: Literature describing the immune response to various patterns of exercise was compared with data on the immune changes observed during sepsis and inflammation. RESULTS: Although there are qualitative similarities between the immune responses to exercise and sepsis, the magnitude of the changes induced by most forms of exercise remains much smaller than in a typical inflammatory response. Indeed, the exercise induced changes in some key elements such as plasma cytokine concentrations are too small to be detected reliably by current technology. CONCLUSIONS: If exercise is to provide a valid model of sepsis and the inflammatory response, it will be necessary to focus on subjects who are willing to exercise extremely hard, to use the pattern of exercise that has the greatest effect on the immune system, and to combine this stimulus with other psychological, environmental, or nutritional stressors. PMID- 11477015 TI - Aerobic and anaerobic power responses to the practice of taekwon-do. AB - BACKGROUND: Practising the martial art of taekwon-do (TKD) has been proposed to have beneficial effects on cardiovascular fitness as well as general physical ability. Furthermore, TKD masters and participants have promoted TKD as a total fitness programme. Research studies substantiating this, however, seem to be lacking, perhaps because TKD is recognised more as a method of self defence than a fitness programme. METHODS: Nineteen TKD practitioners with an average age of 13.8 years and 10.4 months of TKD training experience were recruited to participate. Measurements included resting heart rate, aerobic power, anaerobic power, and anaerobic capacity. RESULTS: Paired t test analysis showed no significant differences in either resting heart rate or aerobic power after training. However, significant differences were observed in anaerobic power and anaerobic capacity (p = 0.05). The increases in anaerobic power and anaerobic capacity were 28% and 61.5% respectively. CONCLUSION: The practice of TKD promotes anaerobic power and anaerobic capacity, but not aerobic power, in male adolescents. PMID- 11477016 TI - Injuries in whitewater kayaking. AB - OBJECTIVE: To provide epidemiological data on whitewater kayaking injuries using a descriptive study. METHODS: A retrospective survey was distributed at whitewater events and club meetings, and made available and advertised on the world wide web, through postings and announcements to newsgroups, related sites, and search engines. Data on sex, age, experience, and ability were collected. Injury data collected included mechanism, activity, difficulty of rapid, and self reported severity. RESULTS: Of the 392 kayaking respondents included in the final analysis, 219 suffered 282 distinct injury events. The number of days spent kayaking per season was the only independent predictor of injury. The overwhelming majority of injuries occurred while the kayaker was still in the boat (87%). Striking an object was the most common mechanism of injury (44%), followed by traumatic stress and overuse (25% each). The most common types of injury were abrasion (25%), tendinitis (25%), contusion (22%), and dislocation (17%). The upper extremity, especially the shoulder, was the most commonly injured area of the body. Although half of injured kayakers sought medical care for their injury, and almost one third missed more than one month of kayaking because of the injury, almost all (96%) reported a complete or good recovery. CONCLUSIONS: Factors relating to likelihood of injury appear to be connected with exposure, namely the number of days a year that the sport was pursued. Except for class V (extreme) kayakers, reports of injuries paralleled the number of participants. Kayakers reported injuries predominantly on rivers that they assessed to be at a level appropriate to their skills. PMID- 11477017 TI - Are synchronised swimmers at risk of amenorrhoea? AB - OBJECTIVE: Synchronised swimming is a sport that shares certain characteristics with other aesthetically pleasing sports such as gymnastics and dance. The purpose of this investigation was to ascertain whether the highest ranked synchronised swimmers in the United Kingdom experience menstrual abnormalities, a common medical problem seen in these related activities. METHODS: Twenty three members of the Great Britain synchronised swimming squad completed a questionnaire on menstrual history. Body composition and VO(2)MAX were measured in the laboratory during regular physiological screening. RESULTS: Three of the 23 subjects were oligomenorrhoeic and none were amenorrhoeic. All were postmenarchal. Mean estimated body fat percentage was 23%, and mean VO(2)MAX was 47.2 ml/kg/min. CONCLUSIONS: It appears that synchronised swimmers in the United Kingdom are relatively protected from menstrual disturbances for reasons that cannot be explained in isolation. PMID- 11477019 TI - An analysis of consultations with the crowd doctors at Glasgow Celtic football club, season 1999-2000. AB - OBJECTIVE: To analyse all clinical presentations to the crowd doctors at Scotland's largest football stadium over the course of one complete season. METHODS: A standard clinical record form was used to document all consultations with the crowd doctors including treatment and subsequent referrals. The relevance of alcohol consumption was assessed. RESULTS: A total of 127 casualties were seen at 26 matches, a mean of 4.88 per match. Twenty casualties were transferred to hospital, including one successfully defibrillated after a cardiac arrest. Alcohol excess was a major contributing factor in 26 cases. CONCLUSIONS: The workload of the crowd doctors was very variable and diverse. The social problem of excessive alcohol consumption contributed considerably to the workload. The provision of medical facilities at football grounds means that attendance there is now one of the least adverse circumstances in which to have a cardiac arrest. The study confirmed previous impressions that more casualties are seen at high profile matches. PMID- 11477021 TI - A profile of patients attending sports medicine clinics. AB - OBJECTIVES: To describe the sociodemographic profile of sports injury patients who attend sports medicine clinics for treatment and to describe their reasons for choosing to attend such clinics. METHOD: Data were collected as part of a fully audited injury surveillance system implemented within sports medicine clinics. The study was conducted within five allied multidisciplinary sports medicine clinics in metropolitan Melbourne, Australia. All patients initially presenting for treatment of a new sports or active recreation injury over the period August 1997 to August 1998 were eligible for this study, irrespective of the practitioner providing the treatment. Data were obtained on 6476 patients. RESULTS: The median age of the patients was 25.4 years (range 6.8--81.6) and most were male (69.8% of cases; 95% confidence interval (95% CI) 68.7 to 70.9). Patients had both professional and non-professional backgrounds and were not just local suburb residents. Many patients had insurance cover for their injury treatment: 59.0% (95% CI 58.6 to 59.4) had some private health insurance and 46.6% (95% CI 45.4 to 47.8) had club/association insurance. The most common reasons for attending a clinic was its location (36.8%; 95% CI 36.5 to 37.1) and referral/recommendation (31.0%; 95% CI 30.7 to 31.3). CONCLUSION: Sports medicine clinics provide treatment for a broad spectrum of injured sports participants across a variety of sporting/recreation contexts. Although these clinics mainly serve the immediate geographic community, the sports speciality and expertise of a particular clinic can attract patients from further afield. This information can help sports medicine clinics to market their services more efficiently to meet the needs of their potential patients. PMID- 11477022 TI - Transient forces generated by projectiles on variable quality mouthguards monitored by instrumented impact testing. AB - OBJECTIVES: (a) To determine the force-time trace that occurs when a spring mounted simulated upper jaw is impacted; (b) to examine if mouthguards of variable quality have significant influence on such force-time traces; (c) to attempt to relate physical events to the profile of the force-time traces recorded. METHODS: A simulated jaw, consisting of ceramic teeth inserted into a hard rubber arch reinforced with a composite jawbone, was fitted with various mouthguards as part of a previous round robin study. A clinical assessment distinguished good, bad, and poor mouthguards, and these were each fitted to the jaw, which was then submitted to instrumental impact tests under conditions expected to produce tooth fractures. The force-time trace was recorded for such impact events. RESULTS: The spring mounting method caused two distinct peaks in the force-time trace. The initial one was related to inertia effects and showed an increase in magnitude with impactor velocity as expected. The second peak showed features that were related to the differences in the mouthguards selected. CONCLUSIONS: The use of a force washer within a conical ended impactor enabled force-time traces to be recorded during the impact of a spring mounted simulated jaw fitted with mouthguards of variable quality. The spring mounting system causes an initial inertial peak followed by a second peak once the spring mount has fully compressed. Good fitting guards, which keep most teeth intact, result in high stiffness targets that in turn generate high reaction forces in the impactor. If the spring mounting is omitted, the two peaks are combined to give even higher reaction forces. The force-time trace offers some potential for assessing both overall mouthguard performance and individual events during the impact sequence. Mouthguards with good retention to the jaw remained attached during the impact event and helped to preserve the structural integrity of the target. This in turn developed high forces in the second part of the force-time trace. Guards that detached during impact and allowed tooth fractures showed lower forces in the second part of the test. The force profile measured offered some quantitative support to, and agreement with, the observed clinical quality of the mouthguards. PMID- 11477024 TI - The reliability and validity of the physical activity questions in the WHO health behaviour in schoolchildren (HBSC) survey: a population study. AB - OBJECTIVE: To assess the test-retest reliability and validity of the physical activity questions in the World Health Organisation health behaviour in schoolchildren (WHO HBSC) survey. METHODS: In the validity study, the Multistage Fitness Test was administered to a random sample of year 8 (mean age 13.1 years; n = 1072) and year 10 (mean age 15.1 years; n = 954) high school students from New South Wales (Australia) during February/March 1997. The students completed the self report instruments on the same day. An independent sample of year 8 (n = 121) and year 10 (n = 105) students was used in the reliability study. The questionnaire was administered to the same students on two occasions, two weeks apart, and test-retest reliability was assessed. Students were classified as either active or inadequately active on their combined responses to the questionnaire items. Kappa and percentage agreement were assessed for the questionnaire items and for a two category summary measure. RESULTS: All groups of students (boys and girls in year 8 and year 10) classified as active (regardless of the measure) had significantly higher aerobic fitness than students classified as inadequately active. As a result of highly skewed binomial distributions, values of kappa were much lower than percentage agreement for test retest reliability of the summary measure. For year 8 boys and girls, percentage agreement was 67% and 70% respectively, and for year 10 boys and girls percentage agreement was 85% and 70% respectively. CONCLUSIONS: These brief self report questions on participation in vigorous intensity physical activity appear to have acceptable reliability and validity. These instruments need to be tested in other cultures to ensure that the findings are not specific to Australian students. Further refinement of the measures should be considered. PMID- 11477025 TI - Physical activity in Dublin children aged 7-9 years. AB - OBJECTIVES: To investigate the amount of regular activity and time spent in sedentary occupations in children aged 7--9 years. Sex differences in levels of activity and time and facilities for physical education at school were also examined. METHODS: A 10% sample of Dublin National Schools were selected. Parents of children in second class were surveyed. The questionnaire used was a modification of the Modifiable Activity Questionnaire for Adolescents. Teachers of second class were questioned about the time and facilities for physical education in schools. RESULTS: Some 39% of children were participating in hard exercise for at least 20 minutes three or more times a week, with fewer girls (28%) than boys (53%) contributing to this result. A further 57% of children were engaging in at least 20 minutes of light exercise three or more times a week, with no sex differences. Estimated energy expenditure in regular activity was higher in boys than girls. Most (78%) of the children were spending one to three hours a day sedentary in front of a screen. CONCLUSIONS: This study provides comprehensive data on physical activity levels in Dublin schoolchildren aged 7--9 years. The amount of inactivity is of concern. Even at this young age, boys are reported to participate in more physical activity than girls. PMID- 11477028 TI - Quantitative heel ultrasound variables in powerlifters and controls. AB - OBJECTIVE: To compare by cross sectional study the quantitative heel ultrasound (QUS) variables broadband ultrasound attenuation (BUA) and velocity of sound (VOS) in male powerlifters and controls. METHODS: Twenty four powerlifters and 21 sedentary male controls were recruited to the study. All the powerlifters were members of the British Drug Free Powerlifting Association and actively competing at the time of the study. A questionnaire was completed by all those entered into the study. This included a history of smoking and an estimation of daily intake of alcohol and calcium. For the powerlifters, the number of years spent training and time spent training each week was also recorded. The QUS variables of all powerlifters and controls were measured using a Cubaclinical II (McCue) ultrasound scanner. RESULTS: The powerlifters had been training for a mean (SEM) of 10.6 (1.6) years and they trained for 6.5 (0.4) hours a week. The powerlifters were non-significantly older and had a significantly higher body mass index (BMI) than the controls. Calcium intake and consumption of alcohol and tobacco were similar in the two groups. The mean BUA in the powerlifters was a significant 9.5% (95% confidence interval 0.7 to 18.3%) higher than the controls (105 v 96 dB/MHZ) and 15.6% (95% confidence interval 6.8 to 24.4%) higher after adjustment using analysis of covariance for age, BMI, and alcohol and tobacco consumption (108 v 93 dB/MHZ). The mean VOS was similar in the two groups, but after adjustment it was significantly higher in the powerlifters (1671 v 1651 m/s, p<0.01). CONCLUSIONS: The study shows the ability of heel ultrasound to discriminate between QUS variables in powerlifters and controls. The results indicate that the QUS variables BUA and VOS are significantly higher for powerlifters than for controls. PMID- 11477029 TI - The kick with the stick. AB - OBJECTIVE: To investigate the incidence and type of severe microscooter related injuries in adults. METHOD: Data were collected between January and September 2000 from the University Hospital Berne, the only referral centre for major trauma in that city, using the software package Qualicare, which connects clinical data with categorised keywords, allowing the immediate localisation of patient groups with defined diagnosis or other clinical information. RESULTS: Only 0.2% of the patients treated had suffered a microscooter accident. There were five head injuries: three facial lacerations, one fractured mandible, and one cerebral concussion. One patient showed clinical signs of a cervical whiplash injury without radiological findings. There were also two cases of finger laceration and two of muscular contusions of the lower extremities. CONCLUSIONS: Although only a small proportion of the trauma cases were the result of riding microscooters, a system of injury surveillance should be started. Furthermore, protective gear should be worn particularly when microscooters are ridden in the street. PMID- 11477030 TI - Sir Percivall Pott--the first sports neurologist? PMID- 11477031 TI - Sports medicine clinics on the NHS: a patient survey. PMID- 11477032 TI - New Zealand Olympic experience--Sydney 2000. PMID- 11477033 TI - The football club doctor system. PMID- 11477034 TI - Management of diabetes at high altitude. PMID- 11477035 TI - Reassessing the need for sport diving medicals. PMID- 11477036 TI - The future for the field of sports medicine. PMID- 11477037 TI - Height of Kilimanjaro. PMID- 11477038 TI - Sports doctors' resuscitation skills under examination--additional facts. PMID- 11477040 TI - Post-normal medicine. PMID- 11477039 TI - Why does primary care need more implementation research? PMID- 11477041 TI - Why do GPs not implement evidence-based guidelines? A descriptive study. AB - BACKGROUND: There is an acknowledged gap between research findings and their implementation in clinical practice despite the existence of effective educational interventions. OBJECTIVES: Our aim was to identify what is impeding GPs from pursuing currently recognized good practice and implementing evidence based guidelines in their management of hypertension in the elderly. METHOD: We carried out a qualitative study using semi-structured interviews conducted during focus group outreach visits to 34 GPs from nine practices in Merseyside involved in an educational programme designed to improve the management of hypertension in the elderly. RESULTS: Several barriers to the implementation of evidence-based guidelines in the management of hypertension in the elderly were identified. These included: doubts about the applicability of trial data to particular patients; the poor adherence of GPs to practice protocols; ageist attitudes of some GPs; the effect of time pressure and financial considerations making the subject a low priority; the absence of an effective computer system; and the absence of an educational mentor. All participants demonstrated a very positive attitude to practice-based education. They also welcomed external audit data, which compared their performance with that of other practices. Single-handed GPs were particularly enthusiastic about this approach as it provided them with the peer pressure they lacked. CONCLUSIONS: In order to bridge the gap between research and practice, educators need to address the various 'barriers to change' amongst practitioners. PMID- 11477042 TI - Promoting evidence-based medicine in general practice-the impact of academic detailing. AB - BACKGROUND: We undertook a project to promote evidence-based medicine (EBM) within a network of GPs (the Monash Division of General Practice) in Melbourne, Australia. A principal promotional strategy was to conduct practice visits ('academic detailing'). OBJECTIVES: The aim of this study was to measure the impact of academic detailing on GP attitudes and knowledge of EBM. METHODS: All 132 GP members of the division were invited by mail to accept a practice visit about EBM. The GPs had been randomized to one of two groups: to receive academic detailing during the study period or to be visited at a later date. The practice visit consisted of a 30- 45 minute discussion about EBM and the barriers to its practice. Pre- and post-intervention questionnaires were used to measure change in knowledge of and attitudes to EBM over a 3-month period in both groups. RESULTS AND CONCLUSIONS: Academic detailing led to a significant improvement in knowledge scores and self-perceived understanding of EBM, but had little influence on GP attitudes toward it. It is not known whether this would lead to change in clinical behaviour among GPs. PMID- 11477043 TI - Perceptions of genetic risk assessment and education among first-degree relatives of colorectal cancer patients and implications for physicians. AB - BACKGROUND: Genetic risk assessment and education is a clinical service that provides an opportunity for individuals with a strong family history of cancer to understand their risk better, identify a screening regimen and discuss benefits and limitations of genetic testing. OBJECTIVES: The aim of this study was to assess knowledge of and attitudes to genetic risk assessment and education among first-degree relatives of patients with colorectal cancer. METHODS: We conducted focus groups among first-degree relatives of patients with colorectal cancer to assess perceptions of genetic risk assessment and education. In the groups, we elicited reactions using two definitions of genetic risk assessment and education one brief and one more detailed-that might be used by a health practitioner during the referral process. RESULTS: Findings revealed a number of misconceptions and concerns including: (i) what is required to prepare for a session and a lack of desire to collect a family history; (ii) what is involved in a session (including assuming that genetic testing is always included in a session); (iii) distrust over accuracy and possible subjectivity of information provided; and (iv) fear of the effect that participation in a session might have on insurance status. CONCLUSIONS: The findings suggest that health practitioners should educate individuals about genetic risk assessment and education during the initial referral process. Further studies should explore how best to do this. PMID- 11477044 TI - The treatment of child and adolescent mental health problems in primary care: a systematic review. AB - BACKGROUND: There is significant potential to increase the accessibility and effectiveness of child and adolescent mental health services through the involvement of primary care professionals and the delivery of interventions in the primary care setting. However, little is known about the actual clinical and cost-effectiveness of such service delivery. OBJECTIVE: The aim of the study was to review systematically the evidence concerning the effectiveness of interventions for child and adolescent mental health problems in primary care, and interventions designed to improve the skills of primary care staff. METHODS: Searches were made of The Cochrane Clinical Trials Register, MEDLINE, PSYCINFO, EMBASE and CINAHL, together with correspondence with subject experts and authors of studies, and checking of references in identified papers. RESULTS AND CONCLUSIONS: There was some preliminary evidence that treatments by specialist staff working in primary care were effective, although the quality of included studies was variable and no data were available on the cost-effectiveness of interventions. Equally, some educational interventions show potential for increasing the skills and confidence of primary care staff, but controlled evaluations were rare and few studies reported actual changes in professional behaviour or patient health outcomes. A significant programme of research is required if the potential for child and adolescent mental health services in primary care is to be realized in an effective and efficient way. PMID- 11477045 TI - New-onset palpitations in general practice: assessing the discriminant value of items within the clinical history. AB - BACKGROUND: Palpitations are non-specific, with less than half of patients experiencing palpitations having a cardiac arrhythmia. Currently it seems that there is little evidence available to assist GPs in discriminating between patients complaining of palpitations who have significant cardiac arrhythmias and those who do not. OBJECTIVES: Our aim was to estimate discriminant functions for specific items of clinical information in relation to the categorization of a patient (aged over 18 years) with a symptom of new-onset palpitations presenting to primary care. METHODS: A network of 62 GPs spread amongst 36 practices agreed to recruit patients with new-onset palpitations over the course of a 9-month study period. Patients consenting to be involved in the study were asked a number of questions, focusing particularly on the medical history, and were requested to complete a Hospital Anxiety and Depression Scale. Each patient was also provided with a RhythmCard cardiac event recorder for up to 2 weeks and was asked to record their heart rhythm if they experienced palpitations. Odds ratios (adjusted for age and sex) were used to compare the clinical information obtained from patients with the final diagnosis. RESULTS: Of the 139 patients with palpitations presenting to GPs, it would appear that males [odds ratio = 2.1 (1.0-4.5)], those with regular palpitations [odds ratio = 2.5 (1.0-5.8)], those experiencing palpitations at work [odds ratio = 3.0 (1.3-7.2)] and those experiencing palpitations affected by sleeping (odds ratio = 3.3 (1.4-7.7)] were more likely to have a cardiac cause for their palpitations. Similar findings were made in an analysis focusing solely on the 81 patients with a RhythmCard result. Furthermore, amongst this group, it is interesting to note that patients with regular palpitations were more than twice as likely to have a 'significant' cardiac arrhythmia as a cause for their palpitations. There were suggestions of dose-response effects between the rate of the palpitation, the duration of the palpitation and the likelihood of it being a 'significant' arrhythmia. CONCLUSIONS: This study provides some information on the characteristics of patients reporting palpitations to GPs who may have 'significant' cardiac arrhythmias. Based on this work, we believe that a larger community-based study would be worthwhile and would provide useful and useable clinical discriminant information for GPs in the settings where they work and amongst the types of patients they encounter. PMID- 11477046 TI - Do GPs sick-list patients to a lesser extent than other physician categories? A population-based study. AB - BACKGROUND: Large differences between physicians in their use of the sickness certification instrument have been described. There are also indications of differences between different categories of physicians, for instance that occupational health physicians and GPs are less generous with certificates of long duration. OBJECTIVES: We therefore decided to test the hypothesis that GPs and occupational health physicians issue more short-term certificates and use partial sick-listing more often than other physicians. METHODS: Certificates for sickness absence during 4 months in 1995 and 2 months in 1996 were collected in eight Swedish counties; a total of 57 563 certificates. From the certificates, a number of variables were extracted. RESULTS: Sickness certificates issued by GPs were on average for a shorter period of time than those issued by other physicians, for the individual certificate as well as for the total sickness period. Occupational health physicians had longer certification periods than GPs but used partial sick-listing more frequently. However, the patients of the various categories of physicians differed regarding age, sex, diagnosis distribution, etc. When the influence of these factors on the duration of the certification periods was taken into account, the GPs still issued significantly shorter periods of sick-leave than the other physicians, followed by the occupational health physicians. CONCLUSIONS: The results may be indicative of a different way of handling the sickness certification instrument among different categories of physicians, especially GPs. PMID- 11477047 TI - Referrals and relationships: in-practice referrals meetings in a general practice. AB - BACKGROUND: GP referrals to secondary care are an important factor in the cost of running the NHS. The known variation in referral rates between doctors has the potential to cause tension within primary care which will be exacerbated by the latest reorganization of primary care and the trend towards capitation-based budgets. The importance of postgraduate learning for GPs has been recognized; continuing professional development is moving towards self-directed practice based learning programmes. Educational interventions have been shown to alter doctors' prescribing behaviour. This, together with the pressure on accounting for referral activity, makes the prospect of improving, and possibly reducing, referral activity through educational interventions very attractive. OBJECTIVES: This study complemented a randomized controlled trial (RCT) which investigated whether an intervention of the type which had reduced prescribing costs would have a similar effect on referral activity. METHODS: The context of the study, description of the characteristics of the practice and the issues seen as important by the doctors and practice manager were identified through preliminary semi-structured interviews. The practice then held a series of educational in practice meetings to discuss referrals and issues arising from referrals. The audio- and videotaped transcripts were interpreted using content and group dynamic analysis. Participants commented upon our preliminary findings. In addition, we used dimensional analysis to induce a preliminary theory describing the effect of the intervention on this general practice which enabled us to review the findings of the parallel RCT. The educational value of the meetings and the learning needs of the participants were also assessed. RESULTS: Our complementary study showed no alteration of practice referral rates following the educational intervention. The qualitative study, unencumbered by the assumptions inherent in the development of the hypothesis tested in the RCT, highlighted the complexity of decision making in general practice and the likely impact of historical background and a variety of internal and external pressures on this self-directive educational intervention. The practice members described the individual and group learning needs identified as a result of the meetings. CONCLUSION: The findings of this study raise important questions for developing practice-based learning. The outcomes of self-directive interventions in practices will be influenced by internal and external events both past and present. Such outcomes may be qualitative and difficult to measure. They are likely to differ from outcomes seen when interventions are applied to groups of doctors who are not all members of the same practice. PMID- 11477048 TI - The EFQM excellence model is useful for primary health care teams. AB - BACKGROUND: Primary care teams are facing an increased need to develop quality programmes at local level. GPs must lead this process and promote a positive organizational culture if they want to achieve and maintain a continuous improvement of the service. OBJECTIVE: The aim of the present study was to test the applicability and reliability of the European Foundation for Quality Management (EFQM) excellence model self-assessment questionnaire in a primary health care organization. METHOD: A cross-sectional study was carried out of the EFQM questionnaire to compare the scores achieved by a primary health care team in Spain caring for 42 000 inhabitants using internal self-assessment with the scores achieved by professional management auditors through an external audit. RESULTS: The scores of each criterion achieved by self-evaluation are similar to or lower than those assessed by the external evaluation. There is agreement in the areas suitable for improvement. CONCLUSIONS: The experience proves the applicability of the EFQM excellence model for primary health care teams and its reliability, at least when the team undergoing self-assessment know they are going to be re-evaluated. There is high concordance in the identification of areas for improvement. PMID- 11477049 TI - A new, brief questionnaire (PEQ) developed in primary health care for measuring patients' experience of interaction, emotion and consultation outcome. AB - BACKGROUND: A deepened understanding of patients' perspectives is essential in order to improve medical communication. By changing focus from patient satisfaction to patient experiences, more immediate, personal and affective responses may be captured. OBJECTIVE: Our aim was to develop a new consultation specific questionnaire on patient experiences. METHODS: The questionnaire was developed in Norwegian primary care in three main phases. Phase 1: focus groups with patients in order to identify important aspects of patients' experiences, and their words and language when describing such experiences. Phase 2: a questionnaire survey with 110 items including 660 patients. Extensive testing resulted in a reduction to 25 items on six dimensions. Phase 3: a questionnaire survey with 25 items including 1092 patients. Psychometric analyses and feedback from patients and physicians involved dimensionality and tests of validity and reliability. RESULTS: A final questionnaire was produced with 18 items on five dimensions: communication; emotions; short-term outcome; barriers; and relations with the auxiliary staff. The validity and reliability estimates were highly satisfactory. Three scales were skewed while two were more equally distributed. Forty-eight per cent of the patients described less than optimal communication experiences; some communication barriers were detected in 70% of the visits and less helpful experiences with the staff were reported in 55% of the visits. Twenty-four per cent of patients left with no positive feelings, and 48% scored low on the outcome scale (knowledge, perceived result). CONCLUSIONS: The patient experience questionnaire (PEQ) emphasizes what patients value the most, i.e. interaction, emotions and outcome, and may represent a valuable tool for doctors who want feedback from their patients on the function of their doctor-patient relationships. PMID- 11477050 TI - Consultations involving people with congenital disabilities: factors that help or hinder giving care. AB - BACKGROUND: Previous studies have suggested that the attitudes of health professionals towards people with disability may be as negative as those of society. Further, even positive attitudes may not always be reflected in the health professional's behaviour. OBJECTIVE: The aim of this study was to examine GPs' (registrars and trainers) consultations with people who have congenital disabilities and to explore incidents when their attitudes were either matched or not matched with their behaviour. METHODS: A purposeful sample of 19 registrars and trainers participated in a semi-structured interview using the critical incident technique. Subjects were asked to describe encounters from their professional life with a person with a congenital disability, when they either had or had not been able to behave as they wished. RESULTS: The results indicated that matching or non-matching between attitudes and behaviour was related to three main themes: aspects of the patient such as their appearance, ease of communication and autonomy; aspects of the GP including their management of personal, expert and professional boundaries and the historical context of the consultation including the GP's personal and professional experience, the familiarity between the GP and the patient and the patient's previous experiences of care. CONCLUSION: The critical incident technique was found to be a useful tool to gain access into this complex and problematic area and the results raise many issues pertinent to the planning of learning opportunities for both undergraduates and postgraduates. PMID- 11477051 TI - Unravelling empowering internal voices-a case study on the interactive use of illness diaries. AB - BACKGROUND: The article is part of a study of an illness diary method for improving clinical communication with patients suffering from long-standing illness without clinical findings. OBJECTIVE: The aim of this study was to demonstrate how patient empowering can be approached through a process of shared insight in a personal illness description. METHODS: This was a single case study from three encounters with a 48-year-old woman suffering from headache, who participated in the illness diary study. Theoretical sampling was used to select the presented case from the sample. The material comprises sections of notes and transcripts from audiotapes. RESULTS: During the encounters, the medical dialogue is changed to include the patient's internal dialogues on her illness and her ways of coping. Her body language is approached and met, and its empowering potential is explored in the dialogue. CONCLUSION: The reflective practitioner may contribute to transform a consultation from repetitive patterns to a dialogue based on the patient's own coping resources. PMID- 11477052 TI - Living with failing lungs: the doctor-patient relationship. AB - OBJECTIVE: The aim of the present study was to explore the perceptions and needs of chronic obstructive pulmonary disease (COPD) patients by a qualitative study using semi-structured interviews. METHODS: Sixteen patients with a diagnosis of COPD (age range 59-75 years) were recruited from two sample frames. One group were under the care of GPs and the second had received in-patient care previously at a district general hospital. Participants had either moderate or severe impairment of respiratory function identified by spirometric testing [forced expiratory volume in 1 second (FEV(1)) <50% of predicted value]. RESULTS: Primary and secondary care samples were chosen in the hope of identifying various aspects of care. However, the patients in this study primarily chose to discuss their relationship with their GP when considering aspects of their lives as a COPD patient. The most significant findings related to the respondents' chronic patient status, their ability to control symptoms and expectations of health care support. All these factors were connected to their perceived relationship with their GP. CONCLUSIONS: Doctors, as gatekeepers of health care resources, were seen to have immense power over the patients' lives. The need to be perceived in a good light by their doctor was intensified by their low self-esteem, poor control of symptoms and their chronic disease status. The majority of patients expressed a need to take a more active part in the decision-making processes involved in the management of their disease. A more balanced doctor-patient relationship could develop if frank and open discussions are based upon a shared decision-making approach. PMID- 11477053 TI - GPs' views of discussions of prognosis in severe COPD. AB - BACKGROUND: Modern palliative care promotes open communication between doctor and patient, which includes access to information about prognosis. GPs play a major role in managing chronic obstructive pulmonary disease (COPD) patients in the final stages of illness. Their views of discussions of prognosis are therefore important if the principles of palliative care are to be extended to COPD. OBJECTIVE: Our aim was to investigate the role that discussions of prognosis play in GPs' management of patients with severe COPD and the factors that influence those discussions. METHODS: We conducted a questionnaire survey of all GP principals of one inner London Health Authority (n = 389) in April 1999. Questionnaire development involved a literature review to identify issues of importance to GPs in the discussion of prognosis in COPD, and in-depth interviews with five GPs. RESULTS: Of the 214 respondents (55% response), 72.5% thought that discussions of prognosis were often necessary or essential in severe COPD. The majority (82%) felt that GPs have an important role in these discussions. However, only a minority (41%) of GPs reported often or always discussing prognosis. Half the GPs were undecided as to whether most patients with COPD wanted to know about their prognosis. Among the GPs who reported rarely or never discussing prognosis (n = 33), a majority felt ill-prepared to discuss the subject (60% reported that there was insufficient information in the primary care notes to be able to discuss prognosis, and 64% found it hard to start discussions with patients). CONCLUSION: Although the majority of GPs acknowledged a need to discuss prognosis in severe COPD, this was not reflected in their reported behaviour. It appears that the palliative care approach of open communication, whilst seen to be relevant to severe COPD, is not applied routinely in managing the disease in primary care. Uncertainty among GPs as to how patients view the discussion of prognosis and inadequate preparation may pose potential barriers. PMID- 11477054 TI - Health, attitude to care and pattern of attendance among gypsy women-a general practice perspective. AB - BACKGROUND: There is a lack of knowledge about health and attitude to care among gypsies. OBJECTIVES: The aim of this study was to explore the reasons for and patterns of attendance among gypsy women in primary health care and to shed light on health problems of gypsies. METHODS: Four gypsy women, frequently attending a primary health care centre, were interviewed in depth. Data were analysed according to grounded theory. Additional facts were received from record files. RESULTS AND CONCLUSIONS: The gypsy women seldom approached the health centre alone but paid a visit together with relatives or friends. The women usually presented the same type of symptoms, often pain, headache and depression, and obtained the same type of diagnosis and treatment. The symptoms had an acute character and the women wanted immediate access. A collective pattern, a hierarchical order and a strict rule system characterized the gypsy life and coloured the relation to health and illness. Young women were especially vulnerable and could easily end up outside the collective and display symptoms. PMID- 11477055 TI - Unwanted pregnancy and contraceptive knowledge: identifying vulnerable groups from a randomized controlled trial of educational interventions. AB - OBJECTIVES: The aim of this study was to identify predictors of contraceptive pill knowledge and their relationship to educational interventions. METHODS: A total of 636 women attending for a follow-up appointment for repeat prescription of the combined oral contraceptive pill with a GP or practice nurse were randomized to receive leaflets (simple summary leaflet or FPA leaflet), advice or neither. Sociodemographic details and contraceptive knowledge were determined using a validated contraceptive knowledge questionnaire sent after 3 months by post. The main outcomes were sociodemographic, contraceptive, attitudinal and educational predictors of knowledge. RESULTS: A total of 522 (82%) had complete questionnaires. After controlling for educational intervention and other confounding variables, independent predictors of knowledge were further education (adjusted odds ratio 2.98, 95% confidence interval 1.78-4.99); number of years on the pill (0-5, 6-10, >10 years) 1.0, 0.56 (0.33-0.95) and 0.34 (0.19-0.59), respectively; past emergency contraception (1.87, 1.18-2.97); and importance attached to not falling pregnant (1.83, 1.02-3.29). These predictors are less powerful than the impact of most educational interventions (range of odds ratios for interventions: 1.85-6.81), and there was no evidence of a separate effect of educational intervention in any subgroup, except that leaflets have a larger effect in women who have needed emergency contraception in the past (no past use or simple summary and FPA leaflets, 1.74 and 0.90, respectively; with past use, 3.47 and 3.83; interaction term chi-square 6.92, P = 0.03). CONCLUSION: Educational interventions are as important as sociodemographic features in determining knowledge. With limited time for full educational interventions in practice, priorities for intervention should be women who have used emergency contraception in the past-who will benefit most-and those on the pill for >5 years or with no further education who are at highest risk due to poor knowledge. PMID- 11477056 TI - GP survey response rate: a miscellany of influencing factors. AB - BACKGROUND: GP response to surveys is acknowledged to vary widely. The minimization of non-response bias and the generalizability of findings are fundamental research issues. OBJECTIVE: The aim of this study was to identify the factors that had influenced GPs' decisions to complete a questionnaire. METHODS: A short questionnaire eliciting GPs' views on minor ailment consultations was sent to 759 GPs from eight English health authorities. The response rate was 54.5%. Factors perceived by respondents to have influenced their decision to complete this questionnaire were also assessed. Subsequently, a feedback summary, together with a short evaluation form, was sent to those GPs requesting it. RESULTS: The response from GPs located in the London area was significantly lower than that from those elsewhere. Respondents identified questionnaire length and the originating institution as the two major factors influencing their decision to return the survey. A single mailing of the evaluation form yielded a response rate of >60% predominantly positive comments. CONCLUSION: Many factors influence a GP's decision to complete a survey. The effect of location has, to date, largely been ignored. Furthermore, this study suggested feedback to be an important issue. Within health services research, all possible factors need to be considered to maximize response, reduce non-response bias and ultimately facilitate the dissemination of findings. PMID- 11477057 TI - Selections from current literature. Colorectal cancer screening. PMID- 11477059 TI - Health services can be cool: partnership with adolescents in primary care. PMID- 11477060 TI - Encouraging adherence to antiretroviral drug regimes. PMID- 11477061 TI - General practice perspective on cancer services. PMID- 11477062 TI - The role of opportunistic vaccination should not be forgotten. PMID- 11477064 TI - The tumor suppressor PTEN positively regulates macroautophagy by inhibiting the phosphatidylinositol 3-kinase/protein kinase B pathway. AB - The tumor suppressor PTEN is a dual protein and phosphoinositide phosphatase that negatively controls the phosphatidylinositol (PI) 3-kinase/protein kinase B (Akt/PKB) signaling pathway. Interleukin-13 via the activation of the class I PI 3-kinase has been shown to inhibit the macroautophagic pathway in the human colon cancer HT-29 cells. Here we demonstrate that the wild-type PTEN is expressed in this cell line. Its overexpression directed by an inducible promoter counteracts the interleukin-13 down-regulation of macroautophagy. This effect was dependent upon the phosphoinositide phosphatase activity of PTEN as determined by using the mutant G129E, which has only protein phosphatase activity. The role of Akt/PKB in the signaling control of interleukin-13-dependent macroautophagy was investigated by expressing a constitutively active form of the kinase ((Myr)PKB). Under these conditions a dramatic inhibition of macroautophagy was observed. By contrast a high rate of autophagy was observed in cells expressing a dominant negative form of PKB. These data demonstrate that the signaling control of macroautophagy overlaps with the well known PI 3-kinase/PKB survival pathway and that the loss of PTEN function in cancer cells inhibits a major catabolic pathway. PMID- 11477065 TI - The gp91phox component of NADPH oxidase is not the voltage-gated proton channel in phagocytes, but it helps. AB - During the "respiratory burst," the NADPH oxidase complex of phagocytes produces reactive oxygen species that kill bacteria and other invaders (Babior, B. M. (1999) Blood 93, 1464-1476). Electron efflux through NADPH oxidase is electrogenic (Henderson, L. M., Chappell, J. B., and Jones, O. T. G. (1987) Biochem. J. 246, 325-329) and is compensated by H(+) efflux through proton channels that reportedly are contained within the gp91(phox) subunit of NADPH oxidase. To test whether gp91(phox) functions as a proton channel, we studied H(+) currents in granulocytes from X-linked chronic granulomatous disease patients lacking gp91(phox) (X-CGD), the human myelocytic PLB-985 cell line, PLB 985 cells in which gp91(phox) was knocked out by gene targeting (PLB(KO)), and PLB-985 knockout cells re-transfected with gp91(phox) (PLB(91)). H(+) currents in unstimulated PLB(KO) cells had amplitude and gating kinetics similar to PLB(91) cells. Furthermore, stimulation with the phorbol ester phorbol 12-myristate 13 acetate increased H(+) currents to a similar extent in X-CGD, PLB(KO), and PLB(91) cells. Thus, gp91(phox) is not the proton channel in unstimulated phagocytes and does not directly mediate the increase of proton conductance during the respiratory burst. Changes in H(+) channel gating kinetics during NADPH oxidase activity are likely crucial to the activation of H(+) flux during the respiratory burst. PMID- 11477066 TI - Chromaffin cell F-actin disassembly and potentiation of catecholamine release in response to protein kinase C activation by phorbol esters is mediated through myristoylated alanine-rich C kinase substrate phosphorylation. AB - The large majority of chromaffin vesicles are excluded from the plasma membrane by a cortical F-actin network. Treatment of chromaffin cells with phorbol 12 myristate 13-acetate produces disassembly of cortical F-actin, increasing the number of vesicles at release sites (Vitale, M. L., Seward, E. P., and Trifaro, J. M. (1995) Neuron 14, 353-363). Here, we provide evidence for involvement of myristoylated alanine-rich protein kinase C substrate (MARCKS), a protein kinase C substrate, in chromaffin cell secretion. MARCKS binds and cross-links F-actin, the latter is inhibited by protein kinase C-induced MARCKS phosphorylation. MARCKS was found in chromaffin cells by immunoblotting. MARCKS was also detected by immunoprecipitation. In intact or permeabilized cells MARCKS phosphorylation increased upon stimulation with 10(-7) m phorbol 12-myristate 13-acetate. This was accompanied by cortical F-actin disassembly and potentiation of secretion. MARCKS phosphorylation, cortical F-actin disassembly, and potentiation of Ca(2+) evoked secretion were inhibited by a peptide (MARCKS phosphorylation site domain sequence (MPSD)) with amino acid sequence corresponding to MARCKS phosphorylation site. MPSD was phosphorylated in the process. A similar peptide (alanine substituted phosphorylated site domain) with four serine residues of MPSD substituted by alanines was ineffective. These results provide the first evidence for MARCKS involvement in chromaffin cell secretion and suggest that regulation of cortical F-actin cross-linking might be involved in this process. PMID- 11477067 TI - A plant-specific cyclin-dependent kinase is involved in the control of G2/M progression in plants. AB - Cyclin-dependent kinases (CDKs) control the key transitions in the eukaryotic cell cycle. All the CDKs known to control G(2)/M progression in yeast and animals are distinguished by the characteristic PSTAIRE motif in their cyclin-binding domain and are closely related. Higher plants contain in addition a number of more divergent non-PSTAIRE CDKs with still obscure functions. We show that a plant-specific type of non-PSTAIRE CDKs is involved in the control of the G(2)/M progression. In synchronized tobacco BY-2 cells, the corresponding protein, accumulated in a cell cycle-regulated fashion, peaking at the G(2)/M transition. The associated histone H1 kinase activity reached a maximum in mitosis and required a yet unidentified subunit to be fully active. Down-regulation of the associated kinase activity in transgenic tobacco plants using a dominant-negative mutation delayed G(2)/M transition. These results provide the first evidence that non-PSTAIRE CDKs are involved in the control of the G(2)/M progression in plants. PMID- 11477068 TI - Characterization of SH2D1A missense mutations identified in X-linked lymphoproliferative disease patients. AB - X-linked lymphoproliferative disease (XLP) is a primary immunodeficiency characterized by extreme susceptibility to Epstein-Barr virus. The XLP disease gene product SH2D1A (SAP) interacts via its SH2 domain with a motif (TIYXXV) present in the cytoplasmic tail of the cell-surface receptors CD150/SLAM, CD84, CD229/Ly-9, and CD244/2B4. Characteristically, the SH2D1A three-pronged interaction with Tyr(281) of CD150 can occur in absence of phosphorylation. Here we analyze the effect of SH2D1A protein missense mutations identified in 10 XLP families. Two sets of mutants were found: (i) mutants with a marked decreased protein half-life (e.g. Y7C, S28R, Q99P, P101L, V102G, and X129R) and (ii) mutants with structural changes that differently affect the interaction with the four receptors. In the second group, mutations that disrupt the interaction between the SH2D1A hydrophobic cleft and Val +3 of its binding motif (e.g. T68I) and mutations that interfere with the SH2D1A phosphotyrosine-binding pocket (e.g. C42W) abrogated SH2D1A binding to all four receptors. Surprisingly, a mutation in SH2D1A able to interfere with Thr -2 of the CD150 binding motif (mutant T53I) severely impaired non-phosphotyrosine interactions while preserving unaffected the binding of SH2D1A to phosphorylated CD150. Mutant T53I, however, did not bind to CD229 and CD224, suggesting that SH2D1A controls several critical signaling pathways in T and natural killer cells. Because no correlation is present between identified types of mutations and XLP patient clinical presentation, additional unidentified genetic or environmental factors must play a strong role in XLP disease manifestations. PMID- 11477069 TI - Evidence for a role of the JNK cascade in Smad7-mediated apoptosis. AB - Smad proteins are central mediators of the transcriptional effects of transforming growth factor beta (TGF-beta) superfamily that regulate a wide variety of biological processes. Smad7, an inhibitory Smad protein that prevents TGF-beta signaling by interacting with the activated type I TGF-beta receptor, was recently shown to induce sensitization of cells to different forms of cell death. Here we examined the effect of Smad7 on the c-Jun N-terminal kinase (JNK) cascade and investigated the role of this cascade in both the inhibitory and apoptotic functions of Smad7. The transient and stable expression of Smad7 caused a strong and sustained activation of JNK. Expression of a dominant-interfering mutant of mitogen-activated protein kinase kinase 4, which completely abolished Smad7-induced activation of JNK, had no effect on Smad7-mediated inhibition of TGF-beta signaling, indicating that the inhibitory function of Smad7 is independent of the JNK cascade. In contrast, expression of the dominant interfering mutant of mitogen-activated protein kinase kinase 4 impaired the ability of Smad7 to promote cell death. These experiments reveal a novel link between Smad7 and the JNK cascade, which is essential for potentiation of cell death by this inhibitory Smad. PMID- 11477070 TI - DJ-1 positively regulates the androgen receptor by impairing the binding of PIASx alpha to the receptor. AB - DJ-1 was first identified as a novel candidate of the oncogene product that transformed mouse NIH3T3 cells in cooperation with an activated ras. Later DJ-1 was also found to be an infertility-related protein that was reduced in rat sperm treated with sperm toxicants that cause infertility in rats. To determine the functions of DJ-1, cDNAs encoding DJ-1-binding proteins were screened by the yeast two-hybrid method. Of several proteins identified, PIASx alpha/ARIP3, a modulator of androgen receptor (AR), was first characterized as the DJ-1-binding protein in this study. DJ-1 directly bound to the AR-binding region of PIASx alpha by an in vitro coimmunoprecipitation assay and also bound to PIASx alpha in human 293T cells. Both proteins were co-localized in the nuclei. PIASx alpha inhibited the AR transcription activity in a dose-dependent manner in cotransfected monkey CV1 cells with an androgen responsive element-luciferase reporter. Introduction of DJ-1 into CV1 cells in a state of inhibition of AR activity by PIASx alpha restored AR transcription activity by absorbing PIASx alpha from the AR-PIASx alpha complex, while a DJ-1 mutant harboring an amino acid substitution at number 130 from lysine to arginine did not restore it. These results indicate that DJ-1 is a positive regulator of the androgen receptor. PMID- 11477071 TI - Characterization of the physical interaction between estrogen receptor alpha and JUN proteins. AB - Activated estrogen receptor alpha (ERalpha) modulates transcription triggered by the transcription factor activator protein-1 (AP-1), which consists of Jun-Jun homodimers and Jun-Fos heterodimers. Previous studies have demonstrated that the interference occurs without binding of ERalpha to DNA but probably results from protein.protein interactions. However, involvement of a direct interaction between ERalpha and AP-1 is still debated. Using glutathione S-transferase pull down assays, we demonstrated that ERalpha bound directly to c-Jun and JunB but not to FOS family members, in a ligand-independent manner. The interaction could occur when c-Jun was bound onto DNA, as shown in a protein-protein-DNA assay. It implicated the C-terminal part of c-Jun and amino acids 259-302 present in the ERalpha hinge domain. ERalpha but not an ERalpha mutant deleted of amino acids 250-303 (ER241G), also associated with c-Jun in intact cells, in the presence of estradiol, as shown by two-hybrid and coimmunoprecipitation assays. We also show that ERalpha, c-Jun, and the p160 coactivator GRIP1 can form a multiprotein complex in vitro and in intact cells and that the ERalpha.c-Jun interaction could be crucial for the stability of this complex. VP16-ERalpha and c-Jun, which both interact with GRIP1, had synergistic effect on GAL4-GRIP1-induced transcription in the presence of estradiol, and this synergistic effect was not observed with the ERalpha mutant VP16-ER241G or when c-Fos, which bound GRIP1 but not ERalpha, was used instead of c-Jun. Finally, ER241G was inefficient for regulation of AP-1 activity, and an ERalpha truncation mutant encompassing the hinge domain had a dominant negative effect on ERalpha action. These results altogether demonstrate that ERalpha can bind to c-Jun in vitro and in intact cells and that this interaction, by stabilizing a multiprotein complex containing p160 coactivator, is likely to be involved in estradiol regulation of AP-1 responses. PMID- 11477072 TI - The N-terminal region and the mid-region complex of the integrin beta 2 subunit. AB - In the primary sequence of the integrin beta subunit, the N-terminal region (NTR) and mid-region are separated by the I-like domain. To determine the spatial relationship and functional properties of the integrin beta(2) NTR and mid region, we constructed beta(2)/beta(7) chimeras in which the NTR, I-like domain, and the mid-region of the beta(2) subunit were replaced by those of beta(7). Changing either the beta(2) NTR or mid-region, but not the I-like domain to that of beta(7) did not affect LFA-1 (alpha(L)beta(2)) formation and surface expression. Thus, the specificity of alpha(L)beta(2) pairing is conferred by the I-like domain but not the NTR or mid-region. Using these chimeras, the epitopes of six anti-beta(2) mAbs (H52, 7E4, AZN-L18, AZN-L27, KIM202, and MEM-148) were mapped. All except H52 require both the NTR and mid-region for epitope expression. Since these mAbs have distinct properties in terms of epitope expression and effect on LFA-1 binding to ICAM-1, we conclude that the beta(2) NTR and mid-region interact extensively. Although the I-like domain is located between the NTR and mid-region, its removal does not affect the folding of the beta(2) NTR/mid-region complex because this complex alone can be expressed as a soluble protein and precipitated by the appropriate mAbs. Finally, the mAbs H52 and 7E4, abrogated KIM185- but not Mg/EGTAinduced LFA-1/ICAM-1 binding and the epitope of MEM-148 is expressed on Mg/EGTA-activated but not resting LFA-1. These results suggest that the NTR/mid-region complex is involved in the regulation of LFA-1 function. PMID- 11477073 TI - Stimulation of eukaryotic flap endonuclease-1 activities by proliferating cell nuclear antigen (PCNA) is independent of its in vitro interaction via a consensus PCNA binding region. AB - Interaction between human flap endonuclease-1 (hFEN-1) and proliferating cell nuclear antigen (PCNA) represents a good model for interactions between multiple functional proteins involved in DNA metabolic pathways. A region of 9 conserved amino acid residues (residues Gln-337 through Lys-345) in the C terminus of human FEN-1 (hFEN-1) was shown to be responsible for the interaction with PCNA. Our current study indicates that 4 amino acid residues in hFEN-1 (Leu-340, Asp-341, Phe-343, and Phe-344) are critical for human PCNA (hPCNA) interaction. A conserved PCNA interaction motif in various proteins from assorted species has been defined as Q(1)X(2)X(3)(L/I)(4)X(5)X(6)F(7)(F/Y)(8), although our results fail to implicate Q(1) (Gln-337 in hFEN-1) as a crucial residue. Surprisingly, all hFEN-1 mutants, including L340A, D341A, F343A, and F344A, retained hPCNA mediated stimulation of both exo- and flap endonuclease activities. Furthermore, our in vitro assay showed that hPCNA failed to bind to the scRad27 (yeast homolog of FEN-1) nuclease. However, its nuclease activities were significantly enhanced in the presence of hPCNA. Four additional Saccharomyces cerevisiae scRad27 mutants, including multiple alanine mutants and a deletion mutant of the entire PCNA binding region, were constructed to confirm this result. All of these mutants retained PCNA-driven nuclease activity stimulation. We therefore conclude that stimulation of eukaryotic hFEN-1 nuclease activities by PCNA is independent of its in vitro interaction via the PCNA binding region. PMID- 11477074 TI - Peroxisome proliferator-activated receptor gamma ligands differentially modulate muscle cell differentiation and MyoD gene expression via peroxisome proliferator activated receptor gamma -dependent and -independent pathways. AB - The effects of distinct classes of peroxisome proliferator-activated receptor gamma (PPARgamma) ligands on myogenesis and MyoD gene expression were examined in mouse skeletal muscle C2C12 myoblasts. Treatment of C2C12 cells with the PPARgamma ligand, 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2), repressed morphologically defined myogenesis and reduced endogenous mRNA levels of the myogenic differentiation markers MyoD, myogenin, and alpha-actin. In contrast, two synthetic PPARgamma ligands, L-805645 and ciglitazone, exhibited no effects. In transient transfection assays, 15d-PGJ2 specifically inhibited the expression of a MyoD promoter-luciferase reporter gene (MyoDLuc) in a cell type- and promoter-specific manner, indicating that 15d-PGJ2 functions in part by repressing MyoD gene transcription. The inhibition of MyoD gene expression by 15d PGJ2 is mediated by the distal region of the MyoD gene promoter. PPARgamma on its own also inhibited MyoDLuc expression and further augmented the 15d-PGJ2 response. In contrast, L-805645 and ciglitazone did not inhibit MyoDLuc expression on their own but did so in the presence of ectopically expressed PPARgamma. Interestingly, a transdominant inhibitor of PPARgamma (hPPARgamma2Delta500) had no effect on the 15d-PGJ2-dependent repression of MyoDLuc expression but overcame L-805645/PPARgamma-dependent repression. Finally, saturating concentrations of L-805645, which did not affect myogenesis, failed to ablate 15d-PGJ2-mediated repression of the myogenic program. Thus, distinct PPARgamma ligands may repress MyoD gene expression through PPARgamma-dependent and -independent pathways, and 15d-PGJ2 can inhibit the myogenic program independent of its cognate receptor, PPARgamma. PMID- 11477075 TI - Polymorphisms in OATP-C: identification of multiple allelic variants associated with altered transport activity among European- and African-Americans. AB - The human organic anion transporting polypeptide-C (OATP-C) (gene SLC21A6) is a liver-specific transporter importantly involved in the hepatocellular uptake of a variety of endogenous and foreign chemicals. In this study, we demonstrate the presence of multiple functionally relevant single-nucleotide polymorphisms (SNPs) in OATP-C in a population of African- and European-Americans. Moreover, examination of 14 nonsynonymous polymorphisms indicated that genotypic frequencies were dependent on race. Functional assessment of 16 OATP-C alleles in vitro revealed that several variants exhibited markedly reduced uptake of the OATP-C substrates estrone sulfate and estradiol 17beta-d-glucuronide. Specifically, alterations in transport were associated with SNPs that introduce amino acid changes within the transmembrane-spanning domains (T217C (Phe-73 --> Leu), T245C (Val-82 --> Ala), T521C (Val-174 --> Ala), and T1058C (Ile-353 --> Thr)) and also with those that modify extracellular loop 5 (A1294G (Asn-432 --> Asp), A1385G (Asp-462 --> Gly), and A1463C (Gly-488 --> Ala)). Cell surface biotinylation experiments indicated that the altered transport activity of some OATP-C variants was due, in part, to decreased plasma membrane expression. Given the relatively high genotypic frequency of the T521C (14%) transition in European Americans and the G1463C (9%) transversion in African-Americans, SNPs in OATP-C may represent a heretofore unrecognized factor influencing drug disposition. PMID- 11477076 TI - High thermostability and lack of cooperative DNA binding distinguish the p63 core domain from the homologous tumor suppressor p53. AB - The p53 protein is the major tumor suppressor in mammals. The discovery of the p53 homologs p63 and p73 defined a family of p53 members with distinct roles in tumor suppression, differentiation, and development. Here, we describe the biochemical characterization of the core DNA-binding domain of a human isoform of p63, p63-delta, and particularly novel features in comparison with p53. In contrast to p53, the free p63 core domain did not show specific binding to p53 DNA consensus sites. However, glutathione S-transferase-fused and thus dimerized p63 and p53 core domains had similar affinity and specificity for the p53 consensus sites p21, gadd45, cyclin G, and bax. Furthermore, the fold of p63 core was remarkably stable compared with p53 as judged by differential scanning calorimetry (T(m) = 61 degrees C versus 44 degrees C for p53) and equilibrium unfolding ([urea](50%) = 5.2 m versus 3.1 m for p53). A homology model of p63 core highlights differences at a segment near the H1 helix hypothetically involved in the formation of the dimerization interface in p53, which might reduce cooperativity of p63 core DNA binding compared with p53. The model also shows differences in the electrostatic and hydrophobic potentials of the domains relevant to folding stability. PMID- 11477077 TI - Role of cysteine residues in structural stability and function of a transmembrane helix bundle. AB - To study the structural and functional roles of the cysteine residues at positions 36, 41, and 46 in the transmembrane domain of phospholamban (PLB), we have used Fmoc (N-(9-fluorenyl)methoxycarbonyl) solid-phase peptide synthesis to prepare alpha-amino-n-butyric acid (Abu)-PLB, the analogue in which all three cysteine residues are replaced by Abu. Whereas previous studies have shown that replacement of the three Cys residues by Ala (producing Ala-PLB) greatly destabilizes the pentameric structure, we hypothesized that replacement of Cys with Abu, which is isosteric to Cys, might preserve the pentameric stability. Therefore, we compared the oligomeric structure (from SDS-polyacrylamide gel electrophoresis) and function (inhibition of the Ca-ATPase in reconstituted membranes) of Abu-PLB with those of synthetic wild-type PLB and Ala-PLB. Molecular modeling provides structural and energetic insight into the different oligomeric stabilities of these molecules. We conclude that 1) the Cys residues of PLB are not necessary for pentamer formation or inhibitory function; 2) the steric properties of cysteine residues in the PLB transmembrane domain contribute substantially to pentameric stability, whereas the polar or chemical properties of the sulfhydryl group play only a minor role; 3) the functional potency of these PLB variants does not correlate with oligomeric stability; and 4) acetylation of the N-terminal methionine has neither a functional nor a structural effect in full-length PLB. PMID- 11477078 TI - Isolation and characterization of subnuclear compartments from Trypanosoma brucei. Identification of a major repetitive nuclear lamina component. AB - Protozoan parasites of the order Kinetoplastida are responsible for a significant proportion of global morbidity and economic hardship. These organisms also represent extremely distal points within the Eukarya, and one such organism, Trypanosoma brucei, has emerged as a major system for the study of evolutionary cell biology. Significant technical challenges have hampered the full exploitation of this organism, but advances in genomics and proteomics provide a novel approach to acquiring rapid functional data. However, the vast evolutionary distance between trypanosomes and the higher eukaryotes presents significant problems with functional assignment based on sequence similarity, and frequently homologues cannot be identified with sufficient confidence to be informative. Direct identification of proteins in isolated organelles has the potential of providing robust functional insight and is a powerful approach for initial assignment. We have selected the nucleus of T. brucei as a first target for protozoan organellar proteomics. Our purification methodology was able to reliably provide both nuclear and subnuclear fractions. Analysis by gel electrophoresis, electron microscopy, and immunoblotting against trypanosome subcellular markers indicated that the preparations are of high yield and purity, maintain native morphology, and are well resolved from other organelles. Minor developmental differences were observed in the nuclear proteome for the bloodstream and procyclic stages, whereas significant morphological alterations were visible. We demonstrate by direct sequencing that the NUP-1 nuclear envelope antigen is a coiled coil protein, containing approximately 20 near-perfect copies of a 144-amino acid sequence. Immunoelectron microscopy localized NUP-1 to the inner face of the nuclear envelope, suggesting that it is a major filamentous component of the trypanosome nuclear lamina. PMID- 11477079 TI - Functionally separate intracellular Ca2+ stores in smooth muscle. AB - In smooth muscle, release via the inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3)R) and ryanodine receptors (RyR) on the sarcoplasmic reticulum (SR) controls oscillatory and steady-state cytosolic Ca(2+) concentrations ([Ca(2+)](c)). The interplay between the two receptors, itself determined by their organization on the SR, establishes the time course and spatial arrangement of the Ca(2+) signal. Whether or not the receptors are co-localized or distanced from each other on the same store or whether they exist on separate stores will significantly affect the Ca(2+) signal produced by the SR. To date these matters remain unresolved. The functional arrangement of the RyR and Ins(1,4,5)P(3)R on the SR has now been examined in isolated single voltage-clamped colonic myocytes. Depletion of the ryanodine-sensitive store, by repeated application of caffeine, in the presence of ryanodine, abolished the response to Ins(1,4,5)P(3), suggesting that Ins(1,4,5)P(3)R and RyR share a common Ca(2+) store. Ca(2+) release from the Ins(1,4,5)P(3)R did not activate Ca(2+)-induced Ca(2+) release at the RyR. Depletion of the Ins(1,4,5)P(3)-sensitive store, by the removal of external Ca(2+), on the other hand, caused only a small decrease ( approximately 26%) in caffeine-evoked Ca(2+) transients, suggesting that not all RyR exist on the common store shared with Ins(1,4,5)P(3)R. Dependence of the stores on external Ca(2+) for replenishment also differed; removal of external Ca(2+) depleted the Ins(1,4,5)P(3)-sensitive store but caused only a slight reduction in caffeine evoked transients mediated at RyR. Different mechanisms are presumably responsible for the refilling of each store. Refilling of both Ins(1,4,5)P(3) sensitive and caffeine-sensitive Ca(2+) stores was inhibited by each of the SR Ca(2+) ATPase inhibitors thapsigargin and cyclopiazonic acid. These results may be explained by the existence of two functionally distinct Ca(2+) stores; the first expressing only RyR and refilled from [Ca(2+)](c), the second expressing both Ins(1,4,5)P(3)R and RyR and dependent upon external Ca(2+) for refilling. PMID- 11477080 TI - Characterization of the reversible conformational equilibrium in the cytoplasmic domain of human erythrocyte membrane band 3. AB - The cytoplasmic domain of erythrocyte membrane band 3 (cdb3) serves as a center of membrane organization, interacting with such proteins as ankyrin, protein 4.1, protein 4.2, hemoglobin, several glycolytic enzymes, and a tyrosine kinase, p72syk. cdb3 exists in a reversible, pH-dependent conformational equilibrium characterized by large changes in Stokes radius (11 A) and intrinsic fluorescence (2-fold). Based on the crystallographic structure of the cdb3 dimer, we hypothesized that the above conformational equilibrium might involve the movement of flanking peripheral protein binding domains away from a shared dimerization domain. To test this hypothesis, we have mutated both donor (W105L) and acceptor (D316A) residues of a prominent H bond that bridges the above two domains and have examined the effect on the resulting conformational equilibrium. Analysis of the intrinsic fluorescence, Stokes radius, thermal stability, urea stability, and segmental mobility of these mutants reveals that the above H bond is indeed present in the low pH conformation of cdb3 and broken in a higher pH conformation. The data further reveal that cdb3 exists in three native pH dependent conformations and that rupture of the aforementioned H bond occurs only during conversion of the low pH conformation to the mid-pH conformation. Conversion of the mid-pH conformation to the high pH conformation would now appear to involve structural changes primarily in the peripheral protein binding domain. Because ankyrin associates avidly with the low pH conformation of cdb3, ankyrin occupancy should strongly influence this structural equilibrium and thereby affect band 3 and perhaps global membrane properties. PMID- 11477081 TI - Diazaborine treatment of Baker's yeast results in stabilization of aberrant mRNAs. AB - Upon Northern blotting, Saccharomyces cerevisiae that was treated with diazaborine showed aberrant mRNAs that were extended at the 3'-end and terminated at secondary processing sites. These bands were also detected in untreated Deltaupf1, Deltaxrn1, and rat7-1 mutants. This finding demonstrates that the aberrant mRNAs also occur in untreated strains in small quantities and can reach the cytoplasm, where they are normally degraded by Xrn1p. Diazaborine treatment stabilizes these mRNAs. The detection of the aberrant bands in the untreated rat7 1 strain indicates that Rat7 is involved in quality control of RNA. The aberrant mRNAs were not detected after diazaborine treatment of a DRG1-1 mutant. Drg1p, a member of the family of AAA (ATPases associated with a variety of cellular activities) proteins, which are thought to represent specific chaperones, may be involved in the process of unfolding the mRNA-ribonucleoprotein complex or in the recognition of aberrant mRNA molecules in the cytoplasm. PMID- 11477082 TI - Expression of p21Waf1/Cip1 and cyclin D1 is increased in butyrate-resistant HeLa cells. AB - Sodium butyrate induced cell cycle arrest in mammalian cells through an increase in p21Waf1/Cip1, although another study showed that this arrest is related to pRB signaling. We isolated variants of HeLa cells adapted to growth in 5 mm butyrate. One of these variants, clone 5.1, constitutively expressed elevated levels of p21Waf1/Cip1 when incubated in regular growth medium and in the presence of butyrate. Despite this elevated level of p21Waf1/Cip1, the cells continue to proliferate, albeit at a slower rate than parental HeLa cells. Western blot analyses showed that other cell cycle regulatory proteins were not up-regulated to compensate for the elevated expression of p21Waf1/Cip1. However, cyclin D1 was down-regulated by butyrate in HeLa cells but not in clone 5.1. We conclude that continued expression of cyclin D1 allowed clone 5.1 to grow in the presence of butyrate and elevated levels of p21Waf1/Cip1. PMID- 11477083 TI - Identification and functional analysis of two novel mutations in the multidrug resistance protein 2 gene in Israeli patients with Dubin-Johnson syndrome. AB - Dubin-Johnson syndrome (DJS) is an inherited disorder characterized by conjugated hyperbilirubinemia and is caused by a deficiency of the multidrug resistance protein 2 (MRP2) located in the apical membrane of hepatocytes. The aim of this study was to identify the mutations in two previously characterized clusters of patients with Dubin-Johnson syndrome among Iranian and Moroccan Jews and determine the consequence of the mutations on MRP2 expression and function by expression studies. All 32 exons and adjacent regions of the MRP2 gene were screened by polymerase chain reaction and DNA sequencing. Two novel mutations were identified in exon 25. One mutation, 3517A-->T, predicting a I1173F substitution, was found in 22 homozygous Iranian Jewish DJS patients from 13 unrelated families and a second mutation, 3449G-->A, predicting a R1150H substitution, was found in 5 homozygous Moroccan Jewish DJS patients from 4 unrelated families. Use of four intragenic dimorphisms and haplotype analyses disclosed a specific founder effect for each mutation. The mutations were introduced into an MRP2 expression vector by site-directed mutagenesis, transfected into HEK-293 cells, and analyzed by a fluorescence transport assay, immunoblot, and immunocytochemistry. Continuous measurement of probenecid sensitive carboxyfluorescein efflux revealed that both mutations impaired the transport activity of MRP2. Immunoblot analysis and immunocytochemistry showed that MRP2 (R1150H) matured properly and localized at the plasma membrane of transfected cells. In contrast, expression of MRP2 (I1173F) was low and mislocated to the endoplasmic reticulum of the transfected cells. These findings provide an explanation for the DJS phenotype in these two patient groups. Furthermore, the close localization of the two mutations identify this region of MRP2 as important for both activity and processing of the protein. PMID- 11477084 TI - The uptake and degradation of matrix-bound lipoproteins by macrophages require an intact actin Cytoskeleton, Rho family GTPases, and myosin ATPase activity. AB - A key cellular event in atherogenesis is the interaction of macrophages with lipoproteins in the subendothelium. In vivo, these lipoproteins are bound to matrix and often aggregated, yet most cell-culture experiments explore these events using soluble monomeric lipoproteins. We hypothesized that the internalization and degradation of matrix-retained and aggregated low density lipoprotein (LDL) by macrophages may involve the actin-myosin cytoskeleton in a manner that distinguishes this process from the endocytosis of soluble LDL. To explore these ideas, we plated macrophages on sphingomyelinase-aggregated LDL bound to smooth muscle cell-derived matrix in the presence of lipoprotein lipase. The macrophages internalized and degraded the LDL, which was mediated partially by the LDL receptor-related protein. Cytochalasin D and latrunculin A, which block actin polymerization, markedly inhibited the uptake and degradation of matrix-retained LDL but not soluble LDL. Inhibition of Rho family GTPases by Clostridium difficile toxin B blocked the degradation of matrix-retained and aggregated LDL by >90% without any inhibition of soluble LDL degradation. However, specific inhibition of Rho had no effect, suggesting the importance of Rac1 and Cdc42. Degradation of matrix-retained, but not soluble, LDL was also blocked by inhibitors of tyrosine kinase, phosphatidylinositol 3-kinase, and myosin ATPase. These findings define fundamental cytoskeletal pathways that may be involved in macrophage foam cell formation in vivo but have been missed by the use of previous cell culture models. PMID- 11477085 TI - Regulation of p53 sequence-specific DNA-binding by covalent poly(ADP ribosyl)ation. AB - We have characterized the covalent poly(ADP-ribosyl)ation of p53 using an in vitro reconstituted system. We used recombinant wild type p53, recombinant poly(ADP-ribose) polymerase-1 (PARP-1) (EC ), and betaNAD(+). Our results show that the covalent poly(ADP-ribosyl)ation of p53 is a time-dependent protein poly(ADP-ribosyl)ation reaction and that the addition of this tumor suppressor protein to a PARP-1 automodification mixture stimulates total protein-poly(ADP ribosyl)ation 3- to 4-fold. Electrophoretic analysis of the products synthesized indicated that short oligomers predominate early during hetero-poly(ADP ribosyl)ation, whereas longer ADP-ribose chains are synthesized at later times of incubation. A more drastic effect in the complexity of the ADP-ribose chains generated was observed when the betaNAD(+) concentration was varied. As expected, increasing the betaNAD(+) concentration from low nanomolar to high micromolar levels resulted in the slower electrophoretic migration of the p53-(ADP ribose)(n) adducts. Increasing the concentration of p53 protein from low nanomolar (40 nm) to low micromolar (1.0 microm) yielded higher amounts of poly(ADP-ribosyl)ated p53 as well. Thus, the reaction was acceptor protein concentration-dependent. The hetero-poly(ADP-ribosyl)ation of p53 also showed that high concentrations of p53 specifically stimulated the automodification reaction of PARP-1. The covalent modification of p53 resulted in the inhibition of the binding ability of this transcription factor to its DNA consensus sequence as judged by electrophoretic mobility shift assays. In fact, controls carried out with calf thymus DNA, betaNAD(+), PARP-1, and automodified PARP-1 confirmed our conclusion that the covalent poly(ADP-ribosyl)ation of p53 results in the transcriptional inactivation of this tumor suppressor protein. PMID- 11477086 TI - Suppressor of fused negatively regulates beta-catenin signaling. AB - Suppressor of fused (Su(fu)) is a negative regulator of the Hedgehog signaling pathway that controls the nuclear-cytoplasmic distribution of Gli/Ci transcription factors through direct protein-protein interactions. We show here that Su(fu) is present in a complex with the oncogenic transcriptional activator beta-catenin and functions as a negative regulator of T-cell factor (Tcf) dependent transcription. Overexpression of Su(fu) in SW480 (APC(mut)) colon cancer cells in which beta-catenin protein is stabilized leads to a reduction in nuclear beta-catenin levels and in Tcf-dependent transcription. This effect of Su(fu) overexpression can be blocked by treatment of these cells with leptomycin B, a specific inhibitor of CRM1-mediated nuclear export. Overexpression of Su(fu) suppresses growth of SW480 (APC(mut)) tumor cells in nude mice. These observations indicate that Su(fu) negatively regulates beta-catenin signaling and that CRM-1-mediated nuclear export plays a role in this regulation. Our results also suggest that Su(fu) acts as a tumor suppressor. PMID- 11477087 TI - Overexpression of manganese superoxide dismutase prevents alcohol-induced liver injury in the rat. AB - Mitochondria are thought to play a major role in hepatic oxidative stress associated with alcohol-induced liver injury. Thus, the hypothesis that delivery of the mitochondrial isoform of superoxide dismutase (Mn-SOD) via recombinant adenovirus would reduce alcohol-induced liver injury was tested. Rats were given recombinant adenovirus containing Mn-SOD (Ad.SOD2) or beta-galactosidase (Ad.lacZ) and then fed alcohol enterally for 4 weeks. Mn-SOD expression and activity of Ad.SOD2 in liver mitochondria of infected animals was increased nearly 3-fold compared with Ad.lacZ-infected controls. Mitochondrial glutathione levels in Ad.lacZ-infected animals were decreased after 4 weeks of chronic ethanol, as expected, but were unchanged in Ad.SOD2-infected animals. Alanine aminotransferase was elevated significantly by ethanol, an effect that was prevented by Ad.SOD2. Moreover, pathology (e.g. the sum of steatosis, inflammation, and necrosis) was elevated dramatically by ethanol in Ad.lacZ treated rats. This effect was also blunted in animals infected with Ad.SOD2. Neutrophil infiltration was increased about 3-fold in livers from both Ad.lacZ- and Ad.SOD2-infected rats by ethanol treatment. Moreover, ESR-detectable free radical adducts in bile were increased about 8-fold by ethanol. Using (13)C labeled ethanol, it was determined that nearly 60% of total adducts were due to the alpha-hydroxyethyl radical adduct. This increase in radical formation was blocked completely by Ad.SOD2 infection. Furthermore, apoptosis of hepatocytes was increased about 5-fold by ethanol, an effect also blocked by Ad.SOD2. Interestingly, tumor necrosis factor-alpha mRNA was elevated to the same extent in both Ad.lacZ- and Ad.SOD2-infected animals follows ethanol exposure. These data suggest that hepatocyte mitochondrial oxidative stress is involved in alcohol-induced liver damage and likely follows Kupffer cell activation, cytokine production, and neutrophil infiltration. These results also support the hypothesis that mitochondrial oxidant production is a critical factor in parenchymal cell death caused by alcohol. PMID- 11477088 TI - Overexpression of lipoprotein lipase in transgenic rabbits inhibits diet-induced hypercholesterolemia and atherosclerosis. AB - Lipoprotein lipase (LPL) is a key enzyme in the hydrolysis of TG-rich lipoproteins. To elucidate the physiological roles of LPL in lipid and lipoprotein metabolism, we generated transgenic rabbits expressing human LPL. In postheparinized plasma of transgenic rabbits, the human LPL protein levels were about 650 ng/ml, and LPL enzymatic activity was found at levels up to 4-fold greater than that in nontransgenic littermates. Increased LPL activity in transgenic rabbits was associated with as much as an 80% decrease in plasma triglycerides and a 59% decrease in high density lipoprotein-cholesterol. Analysis of the lipoprotein density fractions revealed that increased expression of the LPL transgene resulted in a remarkable reduction in the level of very low density lipoproteins as well as in the level of intermediate density lipoproteins. In addition, LDL cholesterol levels in transgenic rabbits were significantly increased. When transgenic rabbits were fed a cholesterol-rich diet, the development of hypercholesterolemia and aortic atherosclerosis was dramatically suppressed in transgenic rabbits. These results demonstrate that systemically increased LPL activity functions in the metabolism of all classes of lipoproteins, thereby playing a crucial role in plasma triglyceride hydrolysis and lipoprotein conversion, and that overexpression of LPL protects against diet induced hypercholesterolemia and atherosclerosis. PMID- 11477089 TI - Identification of a phorbol ester-responsive element in the interferon-gamma receptor 1 chain gene. AB - Human monocytic leukemia THP-1 cells differentiate into macrophage-like cells when treated with 12-O-tetradecanoylphorbol-13-acetate (TPA). During this process, interferon-gamma (IFN-gamma)-inducible expression of human leukocyte antigen-DR alpha is markedly enhanced. The enhancement of human leukocyte antigen DR alpha expression is at least due to the TPA-dependent induction of the IFN gamma receptor 1 chain and IFN-gamma receptor 2 chain genes. Here we have studied the mechanism of TPA-induced up-regulation of the IFN-gamma receptor 1 chain gene. Reporter gene analyses of 5'-deletion constructs of the IFN-gamma receptor 1 gene (IFNGR1) promoter indicated that the critical region for control of transcription and the TPA-responsive element (TRE) were present in the -128 to 109 base pair (bp) region. We confirmed that this region of the IFNGR1 promoter was responsive to TPA-induced signals by using a reporter construct whose promoter consisted of the -128 to -109 bp fragment and the minimal herpes simplex virus thymidine kinase promoter. Moreover, a supershift assay indicated that Sp1 bound to this TRE in TPA-treated THP-1 cells. These results suggest that in TPA treated cells the binding of Sp1 to the TRE of the IFNGR1 promoter causes the up regulation of this gene. PMID- 11477090 TI - The disintegrins ADAM10 and TACE contribute to the constitutive and phorbol ester regulated normal cleavage of the cellular prion protein. AB - We showed previously that PrPc undergoes constitutive and phorbol ester-regulated cleavage inside the 106-126 toxic domain of the protein, leading to the production of a fragment referred to as N1. Here we show by a pharmacological approach that o-phenanthroline, a general zinc-metalloprotease inhibitors, as well as BB3103 and TAPI, the inhibitors of metalloenzymes ADAM10 (A disintegrin and metalloprotease); and TACE, tumor necrosis factor alpha-converting enzyme; ADAM17), respectively, drastically reduce N1 formation. We set up stable human embryonic kidney 293 transfectants overexpressing human ADAM10 and TACE, and we demonstrate that ADAM10 contributes to constitutive N1 production whereas TACE mainly participates in regulated N1 formation. Furthermore, constitutive N1 secretion is drastically reduced in fibroblasts deficient for ADAM10 whereas phorbol 12,13-dibutyrate-regulated N1 production is fully abolished in TACE deficient cells. Altogether, our data demonstrate for the first time that disintegrins could participate in the catabolism of glycosyl phosphoinositide anchored proteins such as PrPc. Second, our study identifies ADAM10 and ADAM17 as the protease candidates responsible for normal cleavage of PrPc. Therefore, these disintegrins could be seen as putative cellular targets of a therapeutic strategy aimed at increasing normal PrPc breakdown and thereby depleting cells of the putative 106-126 "toxic" domain of PrPc. PMID- 11477091 TI - Toll-like receptor 2 (TLR2) and TLR4 differentially activate human dendritic cells. AB - Toll-like receptors (TLRs) mediate cell activation by various microbial products. Here, we demonstrate that activation of dendritic cells by TLR2 or TLR4 agonists, although it led to comparable activation of NF-kappa B and mitogen-activated protein kinase (MAPK) family members, resulted in striking differences in cytokine and chemokine gene transcription, suggesting that TLR2 and TLR4 signaling is not equivalent. A TLR4 agonist specifically promoted the production of the Th1-inducing cytokine interleukin (IL) 12 p70 and the chemokine interferon gamma inducible protein (IP)-10, which is also associated to Th1 responses. In contrast, TLR2 stimulation failed to induce IL-12 p70 and interferon-gamma inducible protein (IP)-10 but resulted in the release of the IL-12 inhibitory p40 homodimer, producing conditions that are predicted to favor Th2 development. TLR2 stimulation also resulted in preferential induction of IL-8 and p19/IL-23. Involvement of phosphatidylinositol 3-kinase and p38 MAPK in the TLR-mediated induction of several cytokine and chemokine messages was demonstrated using specific inhibitors. Thus, TLRs can translate the information regarding the nature of pathogens into differences in the cytokines and chemokines produced by dendritic cells and therefore may contribute to the polarization of the acquired immune response. PMID- 11477092 TI - Retrovirus-mediated expression of apolipoprotein A-I in the macrophage protects against atherosclerosis in vivo. AB - We have previously reported that the lack of apolipoprotein (apo) E expression by macrophages promotes foam cell formation in vivo. Because transgenic mice overexpressing human apoA-I from the liver (h-apoA-I TgN) are protected from the atherogenesis induced by apoE deficiency, we hypothesized that the presence of apoA-I in the vessel wall could reduce the negative effect of apoE deficiency on lesion growth. To address this issue, we used both retroviral transduction and transgenic approaches to produce in vivo systems where apoA-I is expressed from macrophages. In the retroviral transduction study, apoA-I-deficient (apoA-I(-/-)) mice reconstituted with apoE-deficient (apoE(-/-)) bone marrow cells that were infected with a retroviral vector expressing human apoA-I (MFG-HAI) had 95% lower atherosclerotic lesion area than that of recipients of apoE(-/-) bone marrow cells infected with the parental virus (MFG). To determine whether the protective effect of locally produced apoA-I was due to the lack of systemic apoA-I, we conducted a different experiment using h-apoA-I TgN mice as recipients of apoE(-/ ) bone marrow with or without human apoA-I (driven by a macrophage-specific transgene defined as mphi-AI). Aortic lesion area in apoE(-/-)/mphi-AI --> h-apoA I TgN mice was decreased by 85% compared with apoE(-/-) --> h-apoA-I TgN mice. These data demonstrate that expression of apoA-I from macrophages protects against atherogenesis without affecting plasma apoA-I and high density lipoprotein cholesterol levels. PMID- 11477093 TI - Fidelity of nucleotide incorporation by human mitochondrial DNA polymerase. AB - We have examined the fidelity of polymerization catalyzed by the human mitochondrial DNA polymerase using wild-type and exonuclease-deficient (E200A mutation) forms of recombinant, reconstituted holoenzyme. Each of the four nucleotides bind and incorporate with similar kinetics; the average dissociation constant for ground state binding is 0.8 microm, and the average rate of polymerization is 37 x s(-1), defining a specificity constant kcat/Km = 4.6 x 10(7) x m(-1) x s(-1). Mismatched nucleotides show weaker ground-state nucleotide binding affinities ranging from 57 to 364 microm and slower rates of polymerization ranging from 0.013 to 1.16 x s(-1). The kinetic parameters yield fidelity estimates of 1 error out of 260,000 nucleotides for a T:T mismatch, 3563 for G:T, and 570,000 for C:T. The accessory subunit increases fidelity 14-fold by facilitating both ground-state binding and the incorporation rate of the correct A:T base pair compared with a T:T mismatch. Correctly base-paired DNA dissociates from the polymerase at a rate of 0.02 x s(-1) promoting processive polymerization. Thus, the mitochondrial DNA polymerase catalyzed incorporation with an average processivity of 1850, defined by the ratio of polymerization rate to the dissociation rate (37/0.02) and with an average fidelity of one error in 280,000 base pairs. PMID- 11477094 TI - Exonuclease proofreading by human mitochondrial DNA polymerase. AB - We have examined the ability of the human mitochondrial DNA polymerase to correct errors in DNA sequence using single turnover kinetic methods. The rate of excision of single-stranded DNA ranged from 0.07 to 0.17 x s(-1), depending on the identity of the 3'-base. Excision of the 3'-terminal base from correctly base paired DNA occurred at a rate of 0.05 x s(-1), indicating that the cost of proofreading is minimal, as defined by the ratio of the k(exo) for correctly base paired DNA divided by the rate of forward polymerization (0.05/37 = 0.14%). Excision of duplex DNA containing 1-7 mismatches was biphasic, and the rate and amplitude of the fast phase increased with the number of mismatches, reaching a maximum of 9 x s(-1). We showed that transfer of DNA from the polymerase to the exonuclease active site and back again occurs through an intramolecular reaction, allowing for a complete cycle of reactions for error correction. For DNA containing a buried mismatch (T:T followed by C:G base pairs), the 3' base was removed at a rate of 3 x s(-1). The addition of nucleotide to the reaction that is identical to the 3' base increased the rate of excision 7-fold to 21 x s(-1). We propose that the free nucleotide enhances the rate of transfer of the DNA to the exonuclease active site by interrupting the correct 3' base pair through interaction with the template base. The exonuclease contribution to fidelity is minimal if the calculation is based on hydrolysis of a single mismatch: (k(exo) + k(pol,over))/(k(pol,over)) = 10, but this value increases to approximately 200 when examining error correction in the presence of nucleotides. PMID- 11477095 TI - Analysis of murine Brca2 reveals conservation of protein-protein interactions but differences in nuclear localization signals. AB - In this report, we have analyzed the protein encoded by the murine Brca2 locus. We find that murine Brca2 shares multiple properties with human BRCA2 including its regulation during the cell cycle, localization to nuclear foci, and interaction with Brca1 and Rad51. Murine Brca2 stably interacts with human BRCA1, and the amino terminus of Brca2 is sufficient for this interaction. Exon 11 of murine Brca2 is required for its stable association with RAD51, whereas the carboxyl terminus of Brca2 is dispensable for this interaction. Finally, in contrast to human BRCA2, we demonstrate that carboxyl-terminal truncations of murine Brca2 localize to the nucleus. This finding may explain the apparent inconsistency between the cytoplasmic localization of carboxyl-terminal truncations of human BRCA2 and the hypomorphic phenotype of mice homozygous for similar carboxyl-terminal truncating mutations. PMID- 11477096 TI - A trail of research from lipoic acid to alpha-keto acid dehydrogenase complexes. PMID- 11477097 TI - Transforming growth factor-beta-induced osteoblast elongation regulates osteoclastic bone resorption through a p38 mitogen-activated protein kinase- and matrix metalloproteinase-dependent pathway. AB - Transforming growth factor-beta (TGF-beta) is a powerful modulator of bone metabolism, and both its anabolic and catabolic effects on bone have been described. Here we have tested the hypothesis that TGF-beta-induced changes in osteoblast shape promote bone resorption by increasing the surface area of bone that is accessible to osteoclasts. The addition of TGF-beta1 to MC3T3-E1 cells resulted in cytoskeletal reorganization, augmented expression of focal adhesion kinase, and cell elongation, accompanied by an increase in the area of cell-free substratum. TGF-beta1 also triggered activation of Erk1/2 and p38 mitogen activated protein (MAP) kinase. The p38 MAP kinase inhibitor PD169316, but not an inhibitor of the Erk1/2 pathway, abrogated the effect of TGF-beta1 on cell shape. The matrix metalloproteinase inhibitor GM6001 also interfered with osteoblast elongation. Treatment of MC3T3-E1 cells seeded at confluence onto bone slices to mimic a bone lining cell layer with TGF-beta1 also induced cell elongation and increased pit formation by subsequently added osteoclasts. These effects were again blocked by PD169316 and GM6001. We propose that this novel pathway regulating osteoblast morphology plays an important role in the catabolic effects of TGF-beta on bone metabolism. PMID- 11477098 TI - The Acyl-CoA synthetases encoded within FAA1 and FAA4 in Saccharomyces cerevisiae function as components of the fatty acid transport system linking import, activation, and intracellular Utilization. AB - Exogenous long-chain fatty acids are activated to coenzyme A derivatives prior to metabolic utilization. In the yeast Saccharomyces cerevisiae, the activation of these compounds prior to metabolic utilization proceeds through the fatty acyl CoA synthetases Faa1p and Faa4p. Faa1p or Faa4p are essential for long-chain fatty acid import, suggesting that one or both of these enzymes are components of the fatty acid transport system, which also includes Fat1p. By monitoring the intracellular accumulation of the fluorescent long-chain fatty acid analogue 4,4 difluoro-5-methyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoic acid, long-chain fatty acid transport was shown to be severely restricted in a faa1 Delta faa4 Delta strain. These data established for the first time a mechanistic linkage between the import and activation of exogenous fatty acids in yeast. To investigate this linkage further, oleoyl CoA levels were defined following incubation of wild type and mutant cells with limiting concentrations of exogenous oleate. These studies demonstrated oleoyl CoA levels were reduced to less than 10% wild-type levels in faa1 Delta and faa1 Delta faa4 Delta strains. Defects in metabolic utilization and intracellular trafficking were also found in the fatty acyl-CoA synthetase-deficient strains. The faa1 Delta faa4 Delta strain had a marked reduction in endogenous acyl-CoA pools, suggesting these enzymes play a role in maintenance of endogenous acyl-CoA pools, metabolism and trafficking. In addition, this strain had levels of in vivo beta-oxidation of exogenous oleate reduced 3-fold when compared with the isogenic parent. Northern analyses demonstrated an additional defect in fatty acid trafficking as FAA1 or FAA4 were required for the transcriptional regulation of the genes encoding the peroxisomal enzymes acyl-CoA oxidase (POX1) and medium-chain acyl-CoA synthetase (FAA2). These data support the hypothesis that fatty acyl-CoA synthetase (Faa1p or Faa4p) functions as a component of the fatty acid import system by linking import and activation of exogenous fatty acids to intracellular utilization and signaling. PMID- 11477099 TI - Werner syndrome protein is regulated and phosphorylated by DNA-dependent protein kinase. AB - DNA double-strand breaks (DSBs) are a highly mutagenic and potentially lethal damage that occurs in all organisms. Mammalian cells repair DSBs by homologous recombination and non-homologous end joining, the latter requiring DNA-dependent protein kinase (DNA-PK). Werner syndrome is a disorder characterized by genomic instability, aging pathologies and defective WRN, a RecQ-like helicase with exonuclease activity. We show that WRN interacts directly with the catalytic subunit of DNA-PK (DNA-PK(CS)), which inhibits both the helicase and exonuclease activities of WRN. In addition we show that WRN forms a stable complex on DNA with DNA-PK(CS) and the DNA binding subunit Ku. This assembly reverses WRN enzymatic inhibition. Finally, we show that WRN is phosphorylated in vitro by DNA PK and requires DNA-PK for phosphorylation in vivo, and that cells deficient in WRN are mildly sensitive to ionizing radiation. These data suggest that DNA-PK and WRN may function together in DNA metabolism and implicate WRN function in non homologous end joining. PMID- 11477100 TI - 15-deoxy-Delta 12,14-prostaglandin J2 induces apoptosis of human hepatic myofibroblasts. A pathway involving oxidative stress independently of peroxisome proliferator-activated receptors. AB - Hepatic myofibroblasts (hMFs) play a key role in the development of liver fibrosis associated with chronic liver diseases. Apoptosis of these cells is emerging as a key process in the resolution of liver fibrosis. Here, we examined the effects of cyclopentenone prostaglandins on apoptosis of human hMFs. Cyclopentenone prostaglandins of the J series markedly reduced hMF viability, with 15-deoxy-Delta(12,14)-prostaglandin J2 (15-d-PGJ2) being the most potent. This effect was independent of peroxisome-proliferator-activated receptors (PPARs), because PPARgamma and PPARalpha agonists did not affect hMF cell viability, and PPARgamma, the nuclear receptor for 15-d-PGJ2, was not expressed in hMFs. Moreover, 15-d-PGJ2 did not act via a cell surface G protein-coupled receptor, as shown in guanosine-5'-O-(3-thiotriphosphate) binding assays. Cell death resulted from an apoptotic process, because 15-d-PGJ2-treated hMFs exhibited condensed nuclei, fragmented DNA, and elevated caspase-3 activity. Moreover, the caspase inhibitor Z-Val-Ala-Asp(OCH3)-fluoromethyl ketone blocked the cytotoxic effect of 15-d-PGJ2. The apoptotic effects of 15-d-PGJ2 were reproduced by H2O2 and blocked by the antioxidants N-acetylcysteine (NAC), N-(2 mercapto-propionyl)-glycine (NMPG) and pyrrolidine dithiocarbamate (PDTC). Accordingly, 15-d-PGJ2 generated rapid production of reactive oxygen species in hMFs, via a NAC/NMPG/PDTC-sensitive pathway. In conclusion, 15-d-PGJ2 induces apoptosis of human hMFs via a novel mechanism involving oxidative stress and unrelated to activation of its nuclear receptor PPARgamma. These data underline the antifibrogenic potential of 15-d-PGJ2. PMID- 11477101 TI - Superimposed levels of regulation of the 4-hydroxyphenylacetate catabolic pathway in Escherichia coli. AB - The regulation of the Pg promoter, which controls the expression of the meta operon of the 4-hydroxyphenylacetic acid (4-HPA) catabolic pathway of Escherichia coli W, has been examined through in vivo and in vitro experiments. By using Pg lacZ fusions we have demonstrated that Pg is a promoter only inducible in the stationary phase when cells are grown on glucose as the sole carbon and energy source. This strict catabolite repression control is mediated by the cAMP receptor protein (CRP). This event does not require the presence of the specific HpaR repressor or the 4-HPA permease (HpaX), excluding the involvement of a typical inducer exclusion mechanism. However, the acetic acid excreted in the stationary phase by the cells growing in glucose acts as an overflow metabolite, which can provide the energy to produce cAMP and to adapt the cells rapidly to the utilization of a new less preferred carbon source such as the aromatic compounds. Although Pg is not a final sigma(38)-dependent promoter, it is activated by the global regulator integration host factor (IHF) in the stationary phase of growth. Gel retardation assays have demonstrated that both CRP and IHF simultaneously bind to the Pg upstream region. DNase I footprint experiments showed that cAMP-CRP and IHF binding sites are centered at -61.5 and -103, respectively, with respect to the transcription start site +1 of the Pg promoter. PMID- 11477102 TI - A novel role for interleukin-18 in adhesion molecule induction through NF kappa B and phosphatidylinositol (PI) 3-kinase-dependent signal transduction pathways. AB - Interleukin-18 (IL-18) is a novel proinflammatory cytokine found in serum and joints of patients with rheumatoid arthritis (RA). We studied a novel role for IL 18 in mediating cell adhesion, a vital component of the inflammation found in RA and other inflammatory diseases. We examined the expression of cellular cell adhesion molecules E-selectin, vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1) on endothelial cells and RA synovial fibroblasts using flow cytometry. Adhesion of the monocyte-like cell line HL-60 to endothelial cells was determined by immunofluorescence. IL-18 significantly enhanced ICAM-1 and VCAM-1 expression on endothelial cells and RA synovial fibroblasts. In addition, IL-18 induced E-selectin expression on endothelial cells and promoted the adhesion of HL-60 cells to IL-18-stimulated endothelial cells. Neutralizing anti-VCAM-1 and anti-E-selectin could completely inhibit HL 60 adherence to endothelial cells. IL-18-induced adhesion molecule expression appears to be mediated through nuclear factor kappa B (NF kappa B) and phosphatidyl-inositol 3 kinase (PI 3-kinase) since addition of inhibitors to either NF kappa B (pyrrolidine dithiocarbamate and N-acetyl-l-cysteine) or PI 3 kinase (LY294002) inhibited RA synovial fibroblast VCAM-1 expression by 50 to 60%. Addition of both inhibitors resulted in inhibition of VCAM-1 expression by 85%. In conclusion, the ability of IL-18 to induce adhesion molecule expression on endothelial cells and RA synovial fibroblasts indicates that IL-18 may contribute to RA joint inflammation by enhancing the recruitment of leukocytes into the joint. IL-18 requires NF kappa B as well as PI 3-kinase to induce VCAM-1 on RA synovial fibroblasts, suggesting that there may be two distinct pathways in IL-18-induced adhesion molecule expression. PMID- 11477103 TI - Smad-interacting protein 1 is a repressor of liver/bone/kidney alkaline phosphatase transcription in bone morphogenetic protein-induced osteogenic differentiation of C2C12 cells. AB - Up-regulation of liver/bone/kidney alkaline phosphatase (LBK-ALP) has been associated with the onset of osteogenesis in vitro. Its transcription can be up regulated by bone morphogenetic proteins (BMPs), constitutively active forms of their cognate receptors, or appropriate Smads. The promoter of LBK-ALP has been characterized partially, but not much is known about its transcriptional modulation by BMPs. A few Smad-interacting transcriptional factors have been isolated to date. One of them, Smad-interacting protein 1 (SIP1), belongs to the family of two-handed zinc finger proteins binding to E2-box sequences present, among others, in the promoter of mouse LBK-ALP. In the present study we investigated whether SIP1 could be a candidate regulator of LBK-ALP transcription in C2C12 cells. We demonstrate that SIP1 can repress LBK-ALP promoter activity induced by constitutively active Alk2-Smad1/Smad5 and that this repression depends on the binding of SIP1 to the CACCT/CACCTG cluster present in this promoter. Interestingly, SIP1 and alkaline phosphatase expression domains in developing mouse limb are mutually exclusive, suggesting the possibility that SIP1 could also be involved in the transcriptional regulation of LBK-ALP in vivo. Taken together, these results offer an intriguing possibility that ALP up regulation at the onset of BMP-induced osteogenesis could involve Smad/SIP1 interactions, resulting in the derepression of that gene. PMID- 11477104 TI - Escherichia coli SecA helicase activity is not required in vivo for efficient protein translocation or autogenous regulation. AB - SecA is an essential ATP-driven motor protein that binds to preproteins and the translocon to promote protein translocation across the eubacterial plasma membrane. Escherichia coli SecA contains seven conserved motifs characteristic of superfamily II of DNA and RNA helicases, and it has been shown previously to possess RNA helicase activity. SecA has also been shown to be an autogenous repressor that binds to its translation initiation region on secM-secA mRNA, thereby blocking and dissociating 30 S ribosomal subunits. Here we show that SecA is an ATP-dependent helicase that unwinds a mimic of the repressor helix of secM secA mRNA. Mutational analysis of the seven conserved helicase motifs in SecA allowed us to identify mutants that uncouple SecA-dependent protein translocation activity from its helicase activity. Helicase-defective secA mutants displayed normal protein translocation activity and autogenous repression of secA in vivo. Our studies indicate that SecA helicase activity is nonessential and does not appear to be necessary for efficient protein secretion and secA autoregulation. PMID- 11477105 TI - Binding of Paxillin to the alpha 9 Integrin Cytoplasmic Domain Inhibits Cell Spreading. AB - alpha(9)beta(1) integrin is a member of the beta(1) integrin family, plays an important role in extravasation of neutrophils at sites of acute inflammation, and is required for the normal development of the lymphatic system. The alpha(9) and alpha(4) integrin subunits are most closely related and form a subfamily of integrin alpha subunits. Previously, we have reported that the alpha(4) cytoplasmic domain directly and tightly binds paxillin, an intracellular signaling adaptor molecule. This interaction accounts for some of the unusual functional responses to alpha(4) integrin-mediated cell adhesion, including stimulation of cell migration and inhibition of cell spreading and focal adhesion formation. In the current studies, we have examined the interaction between the alpha(9) cytoplasmic domain and paxillin. Here we report that the alpha(9) cytoplasmic domain binds paxillin directly and tightly and that the alpha(9) paxillin association inhibits cell spreading. We have identified amino acid residues in the alpha(9) cytoplasmic domain, Trp(999) and Trp(1001), that are critical for paxillin binding, and alanine substitution of either Trp(999) or Trp(1001) blocks paxillin binding. Furthermore, these mutations also reverse the effect of the alpha(9) cytoplasmic domain on cell spreading. Thus, the alpha(9) and alpha(4) integrin subunits form a paxillin-binding subfamily of integrin alpha subunits, and direct binding of paxillin to the alpha(9) cytoplasmic domain mediates some of the biological activities of the alpha(9)beta(1) integrin. PMID- 11477106 TI - Direct demonstration of rapid degradation of nuclear sterol regulatory element binding proteins by the ubiquitin-proteasome pathway. AB - Sterol regulatory element-binding proteins (SREBPs) are synthesized as membrane bound precursors and processed to generate transcriptionally active forms. The active SREBPs translocate to the nucleus, induce the expression of responsive genes, and are degraded very rapidly. Treatment with proteasome inhibitors elevates the amount of the endogenous nuclear SREBPs, but not the precursors, in HeLa cells. Nuclear forms of human SREBP-1a (amino acids 1-487) and SREBP-2 (amino acids 1-481), which are transiently expressed in stable Chinese hamster ovary cell lines (CHO-487 and -481), are also stabilized by proteasome inhibitors, suggesting that the nuclear SREBPs are likely to be substrates for the proteasome-dependent proteolysis. The stabilized nuclear SREBPs actively induce the expression of responsive genes including hydroxymethylglutaryl (HMG) CoA synthase, fatty acid synthase, and the low density lipoprotein receptor. The rapid turnover of nuclear SREBP-1a is not affected by the intracellular sterol levels, and the half-life is estimated to be approximately 3 h. The nuclear SREBPs are found conjugated with a polyubiquitin chain. When this conjugation is inhibited by overexpression of mutant ubiquitin that is defective in polyubiquitination, the nuclear SREBPs are partly stabilized and induce the expression of the responsive gene, suggesting that the ubiquitin-conjugated SREBPs are substrates for the proteasome. Taken together, these results demonstrate that the ubiquitin-proteasome system degrades SREBPs and that this system controls the expression of SREBP-responsive genes. PMID- 11477107 TI - The Sp1-like protein BTEB3 inhibits transcription via the basic transcription element box by interacting with mSin3A and HDAC-1 co-repressors and competing with Sp1. AB - Sp1-like proteins are characterized by three conserved C-terminal zinc finger motifs that bind GC-rich sequences found in promoters of numerous genes essential for mammalian cell homeostasis. These proteins behave as transcriptional activators or repressors. Although significant information has been reported on the molecular mechanisms by which Sp1-like activators function, relatively little is known about mechanisms for repressor proteins. Here we report the functional characterization of BTEB3, a ubiquitously expressed Sp1-like transcriptional repressor. GAL4 assays show that the N terminus of BTEB3 contains regions that can act as direct repressor domains. Immunoprecipitation assays reveal that BTEB3 interacts with the co-repressor mSin3A and the histone deacetylase protein HDAC 1. Gel shift assays demonstrate that BTEB3 specifically binds the BTE site, a well characterized GC-rich DNA element, with an affinity similar to that of Sp1. Reporter and gel shift assays in Chinese hamster ovary cells show that BTEB3 can also mediate repression by competing with Sp1 for BTE binding. Thus, the characterization of this protein expands the repertoire of BTEB-like members of the Sp1 family involved in transcriptional repression. Furthermore, our results suggest a mechanism of repression for BTEB3 involving direct repression by the N terminus via interaction with mSin3A and HDAC-1 and competition with Sp1 via the DNA-binding domain. PMID- 11477108 TI - Multiple phosphorylated isoforms of NRL are expressed in rod photoreceptors. AB - NRL, a bZIP transcription factor of the Maf subfamily, interacts with the homeodomain protein CRX and synergistically regulates rhodopsin expression. Here we report that six isoforms of NRL (29-35 kDa) are generated by phosphorylation and expressed specifically in the mammalian retina. The anti-NRL antibody also cross-reacts with a cytosolic 45-kDa protein, which is detected in neuronal tissues but is not encoded by the NRL gene. In both human retinal cell cultures and sections of fetal and adult human retina, NRL is present in the nuclei of developing and mature rods but not cones. We propose that NRL regulates rod photoreceptor-specific gene expression and is involved in rod differentiation. PMID- 11477109 TI - Structure of eicosapentaenoic and linoleic acids in the cyclooxygenase site of prostaglandin endoperoxide H synthase-1. AB - Prostaglandin endoperoxide H synthases-1 and -2 (PGHSs) can oxygenate 18-22 carbon polyunsaturated fatty acids, albeit with varying efficiencies. Here we report the crystal structures of eicosapentaenoic acid (EPA, 20:5 n-3) and linoleic acid (LA, 18:2 n-6) bound in the cyclooxygenase active site of Co(3+) protoporphyrin IX-reconstituted ovine PGHS-1 (Co(3+)-oPGHS-1) and compare the effects of active site substitutions on the rates of oxygenation of EPA, LA, and arachidonic acid (AA). Both EPA and LA bind in the active site with orientations similar to those seen previously with AA and dihomo-gamma-linolenic acid (DHLA). For EPA, the presence of an additional double bond (C-17/C-18) causes this substrate to bind in a "strained" conformation in which C-13 is misaligned with respect to Tyr-385, the residue that abstracts hydrogen from substrate fatty acids. Presumably, this misalignment is responsible for the low rate of EPA oxygenation. For LA, the carboxyl half binds in a more extended configuration than AA, which results in positioning C-11 next to Tyr-385. Val-349 and Ser-530, recently identified as important determinants for efficient oxygenation of DHLA by PGHS-1, play similar roles in the oxygenation of EPA and LA. Approximately 750 and 175-fold reductions in the oxygenation efficiency of EPA and LA were observed with V349A oPGHS-1, compared with a 2-fold change for AA. Val-349 contacts C-2 and C-3 of EPA and C-4 of LA orienting the carboxyl halves of these substrates so that the omega-ends are aligned properly for hydrogen abstraction. An S530T substitution decreases the V(max)/K(m) of EPA and LA by 375- and 140 fold. Ser-530 makes six contacts with EPA and four with LA involving C-8 through C-16; these interactions influence the alignment of the substrate for hydrogen abstraction. Interestingly, replacement of Phe-205 increases the volume of the cyclooxygenase site allowing EPA to be oxygenated more efficiently than with native oPGHS-1. PMID- 11477110 TI - The yeast SEC20 gene is required for N- and O-glycosylation in the Golgi. Evidence that impaired glycosylation does not correlate with the secretory defect. AB - The Golgi plays a fundamental role in posttranslational glycosylation, transport, and sorting of proteins. The mechanism of protein transport through the Golgi has been seen as controversial in recent years. During the characterization of N glycosylation-defective mutants (ngd) previously isolated by this laboratory, it was found that ngd20 is allelic to sec20. SEC20 was reported to be required for transport from endoplasmic reticulum to Golgi, but its precise function remains to be determined. We show now that SEC20 is also required for N- and O glycosylation in the Golgi but not in the ER, in a cargo-specific manner, and that the glycosylation defect does not correlate with the secretory defect. By pulse-chase labeling experiments in combination with mannose linkage-specific antibodies, invertase and carboxypeptidase were found to be efficiently secreted to their final compartment, even upon shift to the nonpermissive temperature, while glycosylation in the Golgi was severely impaired. Using microsomal membranes isolated from ngd20, we found that mannosyl transfer from GDP-Man to various mannose-oligosaccharides, indicative for Golgi mannosylation, was strongly diminished. Analysis of the carbohydrate component of chitinase, an exclusively O-mannosylated protein, or of the bulk mannoprotein indicates that O mannosylation is also reduced. The results demonstrate that in addition to secretion SEC20 also affects glycosylation in the Golgi, presumably because it exerts a more general role in maintenance and function of the Golgi compartments. PMID- 11477112 TI - Benchmarking and the laboratory. AB - This article describes how benchmarking can be used to assess laboratory performance. Two benchmarking schemes are reviewed, the Clinical Benchmarking Company's Pathology Report and the College of American Pathologists' Q-Probes scheme. The Clinical Benchmarking Company's Pathology Report is undertaken by staff based in the clinical management unit, Keele University with appropriate input from the professional organisations within pathology. Five annual reports have now been completed. Each report is a detailed analysis of 10 areas of laboratory performance. In this review, particular attention is focused on the areas of quality, productivity, variation in clinical practice, skill mix, and working hours. The Q-Probes scheme is part of the College of American Pathologists programme in studies of quality assurance. The Q-Probes scheme and its applicability to pathology in the UK is illustrated by reviewing two recent Q Probe studies: routine outpatient test turnaround time and outpatient test order accuracy. The Q-Probes scheme is somewhat limited by the small number of UK laboratories that have participated. In conclusion, as a result of the government's policy in the UK, benchmarking is here to stay. Benchmarking schemes described in this article are one way in which pathologists can demonstrate that they are providing a cost effective and high quality service. PMID- 11477111 TI - New insights into the role of cytokines in asthma. AB - Asthma is a triad of intermittent airway obstruction, bronchial smooth muscle cell hyperreactivity to bronchoconstrictors, and chronic bronchial inflammation. From an aetiological standpoint, asthma is a heterogeneous disease, but often appears as a form of immediate hypersensitivity. Many patients with asthma have other manifestations of atopy, such as rhinitis or eczema. Even among non-atopic patients with asthma, the pathophysiology of airway constriction is similar, raising the hypothesis that alternative mechanisms of mast cell degranulation may underlie the disease. The primary inflammatory lesion of asthma consists of accumulation of CD4(+) T helper type 2 (TH2) lymphocytes and eosinophils in the airway mucosa. TH2 cells orchestrate the asthmatic inflammation through the secretion of a series of cytokines, particularly interleukin 4 (IL-4), IL-13, IL 5, and IL-9. IL-4 is the major factor regulating IgE production by B cells, and is required for optimal TH2 differentiation. However, blocking IL-4 is not sufficient to inhibit the development of asthma in experimental models. In contrast, inhibition of IL-13, another TH2 cytokine whose signal transduction pathway overlaps with that of IL-4, completely blocks airway hyperreactivity in mouse asthma models. IL-5 is a key factor for eosinophilia and could therefore be responsible for some of the tissue damage seen in chronic asthma. IL-9 has pleiotropic activities on allergic mediators such as mast cells, eosinophils, B cells and epithelial cells, and might be a good target for therapeutic interventions. Finally, chemokines, which can be produced by many cell types from inflamed lungs, play a major role in recruiting the mediators of asthmatic inflammation. Genetic studies have demonstrated that multiple genes are involved in asthma. Several genome wide screens point to chromosome 5q31--33 as a major susceptibility locus for asthma and high IgE values. This region includes a cluster of cytokine genes, and genes encoding IL-3, IL-4, IL-5, IL-9, IL-13, granulocyte macrophage colony stimulating factor, and the beta chain of IL-12. Interestingly, for some of these cytokines, a linkage was also established between asthma and their receptor. Another susceptibility locus has been mapped on chromosome 12 in a region that contains other potential candidate cytokine genes, including the gene encoding interferon gamma, the prototypical TH1 cytokine with inhibitory activities for TH2 lymphocytes. Taken together, both experimental and genetic studies point to TH2 cytokines, such as IL-4, IL-13, IL 5, and IL-9, as important targets for therapeutic applications in patients with asthma. PMID- 11477113 TI - Fas ligand upregulation is an early event in colonic carcinogenesis. AB - BACKGROUND/AIMS: Fas ligand (FasL) is a mediator of apoptosis via the Fas receptor (Fas/CD95/APO-1). Normal colonic epithelium expresses Fas, and appears to be relatively sensitive to Fas mediated apoptosis. Colonic adenocarcinomas coexpress FasL and Fas without undergoing widespread apoptosis. This study investigates the expression of FasL in colonic carcinogenesis from the earliest stages of the adenoma-carcinoma sequence. METHODS: FasL expression was determined in colonic adenomas (n = 38) of varying degrees of dysplasia and histological type by immunohistochemistry. Adenomas that contained areas of carcinomatous change were included (n = 12 of 38). Normal colonic epithelium (n = 10), hyperplastic polyps (n = 8), and serrated adenomas (n = 3) from patients without colonic adenocarcinomas were used for comparison. Cell death was detected in situ in adenomas using TUNEL (terminal transferase mediated dUTP nick end labelling). RESULTS: In normal colonic epithelium and hyperplastic polyps, FasL expression was restricted to the luminal surface of the crypts, where Fas-FasL coexpression was coincident with a high frequency of TUNEL positive epithelial cells. All adenomas (n = 38) had an altered distribution of positive FasL staining; FasL expression was found in most cells (> 70% of neoplastic cells). Expression of Fas was also detected throughout the adenomas, but coexpression of FasL and Fas was not associated with TUNEL positivity in most cells. CONCLUSIONS: FasL upregulation occurs early in the adenoma-carcinoma sequence of colon carcinogenesis, and is evident at the level of mild dysplasia. The lack of pronounced apoptosis in areas of adenomas coexpressing Fas and FasL suggests that colonocytes acquire resistance to Fas mediated apoptosis early in the transformation process. PMID- 11477114 TI - A 10 year retrospective comparison of the diagnostic usefulness of synovial fluid and synovial biopsy examination. AB - BACKGROUND/AIMS: Synovial fluid examination is thought to be the pathological investigation of choice in most joint disorders, with only a few specific conditions necessitating biopsy, although no evidence based studies are available to support this belief. This study sought to investigate the validity of this assumption. METHODS: One hundred and three cases in which synovial fluid aspiration and synovial biopsy had both been performed at arthroscopy were studied. The amount of diagnostically useful information produced by each investigation was assessed. RESULTS: In most cases, both investigations provided the same amount of information and were generally equally specific or equally non specific. Overall, the biopsy provided more information than the fluid in 29% of cases and vice versa in 18%. When only those cases in which both tests were adequate were considered, the biopsy provided more specific information than the fluid in a small number (9%) of cases, but these cases could not be predicted. CONCLUSION: The diagnostic usefulness of a biopsy approximates and occasionally exceeds that of a fluid. In the arthroscopic situation, the main advantage of performing both tests is that it provides a "failsafe mechanism" for the rare occasions when one of the samples is inadequate. PMID- 11477115 TI - Erythroleukaemia in the north of England: a population based study. AB - AIMS: To evaluate the incidence and outcome of acute myeloid leukaemia (AML), FAB M6 (erythroleukaemia). METHODS: A demographic study in the Northern Health Region of England between 1983 and 1999. RESULTS: Thirty three cases were diagnosed and registered prospectively. The overall incidence was 0.077 cases/100,000/year. There was a pronounced rise in incidence in patients aged 56 years or more: 6.6 times higher than that in younger patients. Overall survival was poor; median survival was 11 months for those aged less than 56 years, and three months for patients aged 56 years and above (p = 0.045). Acquired karyotypic abnormalities were found in 17 of 27 patients where analysis was attempted. When classified according to the criteria of the Medical Research Council AML trials, karyotype predicted survival, with a median overall survival of 14 months for those with "standard risk" cytogenetic results and two months for "poor risk" results (p = 0.005). CONCLUSION: This study demonstrates a worse survival for patients with erythroleukaemia than that reported in some published trials of selected patients. PMID- 11477116 TI - Identifying sputum specimens of high priority for examination by enhanced mycobacterial detection, identification, and susceptibility systems (EMDISS) to promote the rapid diagnosis of infectious pulmonary tuberculosis. AB - AIMS: To compare clinical information and sputum microscopy as methods for the selection of samples for enhanced mycobacterial detection, identification, and susceptibility systems (EMDISS) to promote the rapid diagnosis of patients with infectious pulmonary tuberculosis. METHODS: Two thousand, two hundred and sixty four specimen request forms were examined for clinical details, which were then used to identify specimens likely to yield Mycobacterium tuberculosis on culture. These results were compared with the results of sputum microscopy for acid fast bacilli (AFB). Both methods were assessed against the results of culture using a combination of continuous automated mycobacterial liquid culture (CAMLiC) and conventional solid culture. RESULTS: Classification based on clinical details was an inefficient method of identifying high priority specimens for EMDISS. Although, when given, clinical details were often consistent, a substantial proportion of specimens arrived with no details. This approach would result in the referral of at least 16% of the workload but lead to the detection by culture of only 46% of the M tuberculosis present within it. In contrast, microscopy for AFB defined a much smaller number of specimens (4.8% of the total), which contained 90% of the M tuberculosis isolates. CONCLUSIONS: Microscopy for AFB is the most efficient method for defining sputum specimens suitable for referral for enhanced mycobacteriological techniques. However, it is essential that the methods used for smear preparation and microscopy are of the highest possible standard, otherwise some patients with infectious pulmonary tuberculosis will be denied, unnecessarily, the benefits of important advances in mycobacteriology. PMID- 11477117 TI - Immunohistochemical assessment of human herpesvirus 8 infection in primary central nervous system large B cell lymphomas. AB - BACKGROUND/AIMS: Primary central nervous system large B cell lymphomas (PCNSLs) are frequently associated with Epstein-Barr virus (EBV) in patients with AIDS and less frequently in those without AIDS. Human herpesvirus 8 (HHV-8) has been detected in these tumours by the polymerase chain reaction (PCR) at low copy number, suggesting its presence in a cell compartment other than the malignant one. The aim of this study was to use immunohistochemistry to assess HHV-8 infection in a series of PCNSLs from patients with and without AIDS. METHODS: The antibody LN53, which reacts with the latent nuclear antigen 1 (LNA1) of HHV-8, was used on tissue sections from 35 patients (17 with and 18 without AIDS) with PCNSL. In addition, DNA was available for PCR (open reading frame 26 (ORF 26), ORF 72, ORF 75) in three patients (two without AIDS, one with AIDS). RESULTS: None of the 35 cases contained either DNA sequences or LNA1 positive cells. CONCLUSIONS: These results confirm the lack of HHV-8 infection in tumour cells of PCNSL. In addition, HHV-8 infected bystander cells do not express LNA1 latent protein in this setting. PMID- 11477118 TI - Bacterial translocation and immunohistochemical measurement of gut immune function. AB - AIMS: The local immune response in the small bowel mucosa might play a role in bacterial translocation (BT). The aim of this study was to quantify immune cells and secretory antibodies in the small bowel mucosa, and relate this to BT as assessed by culture of a mesenteric lymph node. METHODS: Immunohistochemical techniques were used to measure the frequency of plasma cells and IgA and IgM positive cells in the lamina propria and semiquantitatively to assess mucosal surface IgA and IgM values in small bowel specimens obtained from 11 patients in whom positive evidence of BT had been identified in a mesenteric lymph node harvested at the time of laparotomy. These were compared with similar specimens obtained from 11 patients in whom a similar lymph node had yielded no growth. RESULTS: BT was associated with a significantly increased median frequency of plasma cells (p < 0.01) and IgA positive cells (p < 0.05) in the lamina propria. The frequency of IgM positive cells was also higher in these patients, although this difference was not significant. In addition, semiquantitatively scored IgA and IgM concentrations at the mucosal surface were both significantly higher in the patients in whom BT had been identified (p = 0.006 and 0.016, respectively). CONCLUSION: Higher numbers of plasma cells and higher IgA and IgM values are present in the small bowel mucosa of patients in whom BT has been shown to occur, suggesting an increased local immune response. PMID- 11477119 TI - Detection of progesterone receptor forms A and B by immunohistochemical analysis. AB - AIM: The measurement of progesterone receptors (PR) is recommended as part of the clinical management of breast and endometrial cancers, and immunohistochemistry on formalin fixed tissue is now the method of choice. PR is expressed as two isoforms, PRA and PRB, and although both these proteins are expressed in hormone dependent cancers, there is evidence that a large proportion of tumours express a predominance of one isoform. Therefore, it is essential to document the individual detection of PRA and PRB by the presently available anti-PR antibodies. The aim of this study is to investigate the detection of PR isoforms A and B in formalin fixed, paraffin wax embedded cell lines and tissue sections by immunohistochemistry, using a panel of commercial and in house antibodies to human PR. METHODS: PR negative cell lines stably transfected to express only PRA (MCF-7Mll/PRA) or PRB (MDA-MB-231/PRB), and tissue sections of human breast carcinoma and normal endometrium were stained using an immunoperoxidase method. A panel of primary PR specific antibodies was evaluated for ability to detect both PRA and PRB proteins, and for intensity and distribution of positive staining under optimal conditions. RESULTS: Of the 11 antibodies assessed, only four recognised PRA and PRB similarly. Six recognised PRA proteins but were unable to detect PRB expression in the cell lines expressing only PRA or PRB. In tissues expressing high amounts of PRA and PRB, all antibodies tested demonstrated positive PR staining. However, in tissues expressing a predominance of PRB, differential staining patterns were observed, with variations in staining intensity and in the proportion of cells positive for PR. CONCLUSIONS: Most PR specific antibodies tested failed to detect PRB in formalin fixed tissue by immunohistochemical techniques, despite their ability to do so by immunoblot analysis. These observations suggest that there are conformational differences between PRA and PRB that mask epitopes on the PRB protein recognised by most anti PR antibodies. The selection of antibodies that recognise both PRB and PRA in formalin fixed tissue is essential for the accurate evaluation of PR positivity in clinical specimens. PMID- 11477120 TI - Malignant peripheral nerve sheath tumour arising within neurofibroma. An immunohistochemical analysis in the comparison between benign and malignant components. AB - AIMS: To compare the expression of immunohistochemical variables between benign and malignant components of malignant peripheral nerve sheath tumour (MPNST) arising within neurofibroma. METHODS: Eight cases of MPNST arising within a neurofibroma, associated with neurofibromatosis type 1 (NF1), were studied. The areas of MPNST and neurofibroma were compared immunohistochemically with regard to the expression of proliferative activity (MIB-1), growth factors, p53, bcl-2, neural cell adhesion molecule (N-CAM), and CD34. RESULTS: The expression of transforming growth factor beta 1 (TGF-beta 1), TGF-beta receptor type II, hepatocyte growth factor alpha (HGF-alpha), c-met, p53, and N-CAM was higher in the areas of MPNST than in the neurofibromatous areas in four, five, five, eight, five, and three of the eight cases, respectively. CD34 expression was lower in the areas of MPNST than in the neurofibroma areas in three of the eight cases. CONCLUSIONS: On the basis of these findings, TGF-beta 1, HGF-alpha, and p53 might be involved in the malignant transformation of neurofibroma to MPNST. PMID- 11477121 TI - Association of a renal papillary carcinoma with a low grade tumour of the collecting ducts. AB - This case report describes a 75 year old man who had a renal papillary carcinoma associated with a low grade tumour of the collecting ducts. These tumours showed different immunohistochemical patterns for epithelial membrane antigen, cytokeratin 19, and Ulex europaeus lectin expression. In addition, cytogenetic findings were 47, XY, +7 <7> and 45, XY, -8, add(12)(q-ter)<10> for the papillary renal carcinoma and the low grade tumour of the collecting ducts, respectively. This is the first report where these two types of tumour are associated and cytogenetically distinguished. PMID- 11477122 TI - Eosinophilic angiocentric fibrosis and Wegener's granulomatosis: a case report and literature review. AB - This report presents a case of eosinophilic angiocentric fibrosis in a man with Wegener's granulomatosis, the first report of a possible association between the two conditions. This association suggests a possible mechanism for its pathogenesis. PMID- 11477123 TI - Pulmonary marginal zone lymphoma of MALT type as a cause of localised pulmonary amyloidosis. AB - AIM: To describe six patients with pulmonary marginal zone lymphoma in whom amyloid deposition was identified. Marginal zone lymphoma is a recently recognised type of low grade non-Hodgkin's lymphoma. METHODS: A computerised search was performed of all patients seen at the Mayo Clinic with a diagnosis of pulmonary amyloidosis. Six patients with pulmonary amyloidosis who had biopsy confirmed extranodal marginal zone lymphoma of mucosa associated lymphoid tissue type were identified. All were women, ranging in age from 45 to 85 years. RESULTS: Five patients had amyloid deposition in conjunction with pulmonary marginal zone lymphoma at the time of the original diagnosis. One patient was referred for evaluation of localised pulmonary amyloidosis and was found to have coexisting pulmonary marginal zone lymphoma. Clinical presentation was limited to pulmonary symptoms (two of the six) or constitutional symptoms (two), or was asymptomatic (two). In all six cases, initial findings of nodular densities on screening chest roentgenograms led to further evaluation and eventual lobectomy; these findings included multiple pulmonary nodules (four), single nodule (one), and single nodule with diffuse bilateral interstitial infiltrates (one). Bone marrow was examined in five patients and was normal in all. Protein studies performed in four patients revealed no monoclonal protein. No patients had manifestations of systemic amyloidosis, such as renal, neurological, or cardiac involvement, at a median follow up of 50 months. Four of the six patients remain alive at a median of five years. CONCLUSIONS: Pulmonary marginal zone lymphoma may be found in association with localised amyloid deposition and should be considered in the differential diagnosis of localised pulmonary amyloidosis. PMID- 11477124 TI - Workload, organisation, and cost of haematology laboratory out of hours services. AB - BACKGROUND: The cost of out of hours services makes up a considerable proportion of the total laboratory budget and this has encouraged haematologists to examine workload patterns and the organisation and cost of out of hours services. AIM: To review current levels of out of hours workload and examine the organisation and cost of the service in the South Thames region. METHOD: Data collection was by questionnaire and data analysis by Microsoft Excel. RESULTS: A variety of methods of organising and financing out of hours services were identified. There was a large variation in workload between different centres. There has been a rise in total workload and out of hours workload over a three year period. The most common tests to take place outside routine hours are full blood counts and clotting screens. The method of providing out of hours services did not affect total numbers of out of hours tests. There were considerable costs associated with time off in lieu for on call staff. CONCLUSIONS: The variation and increase in out of hours tests is not related to the way that out of hours services are provided, but is more likely to result from changes in medical practice. There are pronounced differences between centres in the numbers of different types of tests performed out of hours, which are not related to the numbers of acute beds. There is no single model of out of hours service provision that suits all situations. PMID- 11477125 TI - How many histological levels should be examined from tissue blocks originating in cone biopsy and large loop excision of the transformation zone specimens of cervix? AB - AIMS: To establish the value of examining additional histological levels in cone biopsy and large loop excision of the transformation zone (LLETZ) specimens of cervix. METHODS: Three deeper levels were examined from 200 consecutive cone biopsy and LLETZ specimens reported by a single pathologist. RESULTS: Examination of the first deeper level resulted in cervical intraepithelial neoplasia (CIN) being identified for the first time in five cases and in CIN1 being upgraded in five more. Invasive cancer was discovered in two cases that had shown high grade CIN initially. CONCLUSION: Examination of a single further level appears to be sufficient in those patients in whom a specimen is compromised because epithelium including the squamocolumnar junction is missing, or if there is a discrepancy between the histological findings and the preceding colposcopic or cytological history. If invasive disease is suspected on the basis of the cytological, colposcopic, or histological features, one or preferably two further levels should be examined. PMID- 11477126 TI - Apolipoprotein E alleles in women with pre-eclampsia. AB - AIMS: To investigate the frequency of three apolipoprotein E (apoE) alleles among women with pre-eclampsia. METHODS: The presence of the three most common apoE alleles (epsilon 2, epsilon 3, epsilon 4) was determined by polymerase chain reaction-restriction fragment length polymorphism in two groups of women: healthy pregnant women (n = 91) and pregnant women with a diagnosis of pre-eclampsia (n = 133). In addition, the frequencies of the alleles in the general population in this area are presented for comparison. RESULTS: The frequency of the apo epsilon 4 allele was 18.4% among women with pre-eclampsia and 18.7% among healthy pregnant women (Fisher's exact test; p = 0.941), which is close to the rate in the general population in this area (19%). None of the apolipoprotein E genotypes was significantly over-represented, and homozygous genotype epsilon 4 was not associated with more severe clinical disease than were the other genotypes. CONCLUSION: The observed profiles of allele and genotype frequencies confirm an equilibrium state between apoE polymorphism and pre-eclampsia and suggest that apoE does not play a major role in the development of pre-eclampsia. PMID- 11477127 TI - Clear cell carcinoma of the ovary arising in a mucinous cystadenoma. PMID- 11477128 TI - Small cell melanoma. PMID- 11477129 TI - Rhabdoid phenotype in cutaneous squamous carcinoma: an earlier report! PMID- 11477130 TI - Molecular pathology of solid tumours: translating research into clinical practice. Introduction and overview. PMID- 11477131 TI - Molecular pathology of solid tumours: some practical suggestions for translating research into clinical practice. AB - "Molecular pathology" can be broadly defined as the use of genetic data, in addition to the standard pathological parameters, to optimise diagnosis and to indicate treatment and prognosis. The benefit to be gained from the exploitation of molecular techniques to provide additional information to aid patient management is potentially vast. Currently, molecular pathology is rarely used in clinical practice, although it is anticipated that it will eventually become a part of routine practice. However, incorporating molecular techniques into routine practice will not be straightforward because there are several issues to be resolved. Following on from a symposium held at the Royal College of Pathologists to discuss some of these issues, the establishment of a committee of molecular pathology is proposed to plan and coordinate the introduction of molecular pathology into routine clinical practice. PMID- 11477132 TI - What we could do now: molecular pathology of colorectal cancer. AB - The contribution of molecular genetics to colorectal cancer has been largely restricted to relatively rare inherited tumours and to the detection of germ line mutations predisposing to these cancers. However, much is now known about the somatic events leading to colorectal cancer in general. Several studies have examined the relation between genetic features and prognosis. The purpose of this article is to review these studies and summarise the current state of this subject. Although many of the published studies are small and inconclusive, it is clear that several different pathways exist for the development of this cancer, and some molecular characteristics seem to correlate with clinicopathological features. At present, studies are confined to evaluating a small number of molecular markers; however, with the advent of methods for the rapid genetic profiling of large numbers of colorectal cancers, it will be possible to evaluate fully the clinical usefulness of a range of colorectal cancer genotypes. PMID- 11477133 TI - What we could do now: molecular pathology of bladder cancer. AB - There is much information on the genetic alterations that contribute to the development of bladder cancer. Because it is hypothesised that the genotype of the cancer cell plays a major role in determining phenotype, this genetic information should impact on clinical practice. To date however, this has not happened. Some of the alterations identified in bladder cancer have clear associations with outcome-for example, mutational inactivation of the cell cycle regulator proteins p53 and the retinoblastoma protein (Rb). However, as single markers, these events have insufficient predictive power to be applied in the management of individual patients. The use of panels of markers is a potential solution to this problem. Examples of suitable panels include those genes/proteins with known impact on specific cell cycle checkpoints or with impact on cellular phenotypes, such as immortalisation, invasion, or metastasis. To evaluate such marker panels, large tumour series will be needed-for example, archival samples from completed clinical trials. The use of these valuable resources will require coordination of sample provision. This might involve central collection and distribution of tissue blocks, sections, or tissue arrays and the provision of patient follow up information to laboratories participating in a study. With the availability of microarray technologies, including cDNA and comparative genomic hybridisation arrays, the transcriptome and genome of transitional cell carcinomas of different phenotypes can be compared and will undoubtedly provide a wealth of information with potential diagnostic and prognostic uses. Although these studies can be initiated using small local tissue collections, high quality collection of fresh tissues from new clinical trials will be crucial for proper evaluation of associations with clinical outcome. Funding for molecular pathological studies to date has been poor. To begin to translate molecular information from the laboratory to the clinic and to make maximum use of valuable urological patient resources in the UK, adequate funding and scientific energy are required. Whereas the latter is not in doubt, present funding for this type of translational research is inadequate. PMID- 11477134 TI - What we could do now: molecular pathology of gynaecological cancer. AB - Gynaecological tumours exemplify many of the molecular paradigms of carcinogenesis. The clinical value of many of the molecular abnormalities present is now being tested and it is likely that the identification of at least some of these will become routine in the near future. This may help to refine diagnosis and guide treatment-for example, therapeutic vaccination for human papillomavirus related disease. PMID- 11477135 TI - Commissioning laboratory services. AB - Scientists often develop techniques that have obvious benefits for patients but then find great difficulty in introducing them into the National Health Service usually because the necessary finance does not seem to be available. This article provides a practical guide. PMID- 11477136 TI - The IGF/IGFBP system in CNS malignancy. AB - The insulin-like growth factor (IGF) system includes IGF-I and IGF-II, the type I and type II IGF receptors, and specific IGF binding proteins (IGFBP-1 to IGFBP 6). These factors regulate both normal and malignant brain growth. Enhanced expression of IGF-I and IGF-II mRNA transcripts has been demonstrated in gliomas, meniningiomas, and other tumours. Abnormal imprinting of IGF-II occurs in gliomas, medulloblastomas, and meningiomas. Both types of IGF receptor are expressed in gliomas and, in particular, the type I IGF receptor appears to be upregulated in malignant brain tissue. Antisense IGF-I receptor mRNA induces an antitumour response, resulting in complete brain tumour regression. Clinical trials for the treatment of brain tumours in humans based on a gene transfer protocol using IGF-I receptor antisense are under way. All six IGFBPs are expressed to a variable extent in brain tumours. High concentrations of IGFBP-2 are found in cerebrospinal fluid from patients with malignant central nervous system tumours; therefore, IGFBP-2 might be a useful marker for these tumours. IGFBP-4 appears to be a negative regulator of tumour proliferation. Both in vitro and in vivo experiments suggest that the IGF system represents an important target for the treatment of malignant central nervous system tumours and the ongoing trials should provide valuable information for future therapeutic approaches. PMID- 11477137 TI - Human glioma cells transformed by IGF-I triple helix technology show immune and apoptotic characteristics determining cell selection for gene therapy of glioblastoma. AB - AIMS: Insulin-like growth factor type I (IGF-I) antisense cellular gene therapy of tumours is based on the following data: rat glioma or hepatoma cells transfected with the vector encoding IGF-I antisense cDNA lose their tumorigenicity and induce a tumour specific immune response involving CD8(+) T cells. Recently, using the IGF-I triple helix approach in studies of tumorigenicity, major histocompatibility complex class I (MHC-I) antigens were demonstrated in rat glioma transfected cells. This study used comparative IGF-I antisense and triple helix technologies in human primary glioma cells to determine the triple helix strategy that would be most appropriate for the treatment of glioblastoma. METHODS: The cells were transfected using the IGF-I triple helix expression vector, pMT-AG, derived from the pMT-EP vector. pMT-AG contains a cassette comprising a 23 bp DNA fragment transcribing a third RNA strand, which forms a triple helix structure within a target region of the human IGF-I gene. Using pMT-EP, vectors encoding MHC-I or B7 antisense cDNA were also constructed. RESULTS: IGF-I triple helix transfected glioma cells are characterised by immune and apoptotic phenomena that appear to be related. The expression of MHC-I and B7 in transfected cells (analysed by flow cytometry) was accompanied by programmed cell death (detected by dUTP fluorescein terminal transferase labelling of nicked DNA and electron microscopic techniques). Cotransfection of these cells with MHC-I and B7 antisense vectors suppressed the expression of MHC-I and B7, and was associated with a pronounced decrease in apoptosis. CONCLUSION: When designing an IGF-I triple helix strategy for the treatment of human glioblastoma, the transfected tumour cells should have the following characteristics: the absence of IGF-I, the presence of both MHC-I and B7 molecules, and signs of apoptosis. PMID- 11477138 TI - Microsatellite instability and mutational analysis of transforming growth factor beta receptor type II gene (TGFBR2) in sporadic ovarian cancer. AB - AIMS: To investigate the possible role of mutations in the transforming growth factor beta receptor type II gene (TGFBRII) in ovarian cancer and its relation to microsatellite instability (MSI), 43 sporadic ovarian tumours were analysed for mutations over the entire coding region of the TGFBRII gene. METHODS: Mutational analysis was performed using the polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP) gel analysis, and direct sequencing. MSI analysis included both mononucleotide and dinucleotide microsatellite markers used for radiolabelling and gene scanning. RESULTS: No pathogenic mutations were detected, although sequencing of the polyadenine (poly A) tract in exon 3 using conventional techniques revealed a spurious frameshift mutation that was not present in the same samples analysed using a proofreading Taq polymerase. MSI analysis demonstrated an MSI negative phenotype in 40 of the 43 tumours. None of the three MSI positive tumours demonstrated MSI for mononucleotide markers only. CONCLUSIONS: These findings suggest that: (1) MSI (both conventional and mononucleotide) is infrequent in ovarian cancer and (2) inactivation of the MSH2, MLH1, and MSH6 mismatch repair genes and TGFBR2 gene mutations do not play a major role in ovarian cancer tumorigenesis. The spurious TGFBR2 frameshift mutations detected by sequencing after conventional PCR underline the importance of confirming putative mutations in repetitive sequences by alternative methods. PMID- 11477139 TI - Identification of slide coagulase positive, tube coagulase negative Staphylococcus aureus by 16S ribosomal RNA gene sequencing. AB - AIMS: To ascertain the clinical importance of a strain of slide coagulase positive but tube coagulase negative Staphylococcus species isolated from the blood culture of a 43 year old patient with refractory anaemia with excessive blasts in transformation who had neutropenic fever. METHODS: The isolate was investigated phenotypically by standard biochemical methods using conventional biochemical tests and two commercially available systems, the Vitek (GPI) and API (Staph) systems. Genotypically, the 16S ribosomal RNA (rRNA) gene of the bacteria was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank by multiple sequence alignment. RESULTS: Conventional biochemical tests did not reveal a pattern resembling a known Staphylococcus species. The Vitek system (GPI) showed that it was 94% S. simulans and 3% S. haemolyticus, whereas the API system (Staph) showed that it was 86.8% S. aureus and 5.1% S. warneri. 16S rRNA gene sequencing showed that there was a 0 base difference between the isolate and S. aureus, 28 base difference between the isolate and S. lugdunensis, 39 base difference between the isolate and S. schleiferi, 21 base difference between the isolate and S. haemolyticus, 41 base difference between the isolate and S. simulans, and 23 base difference between the isolate and S. warneri, indicating that the isolate was a strain of S. aureus. Vancomycin was subsequently prescribed and blood cultures taken four days after the start of treatment were negative. CONCLUSIONS: 16S rRNA gene sequencing was useful in ascertaining the clinical importance of the strain of slide coagulase positive but tube coagulase negative Staphylococcus species isolated from blood culture and allowing appropriate management. PMID- 11477140 TI - Nitric oxide synthase immunoreactivity in human bladder carcinoma. AB - AIMS: To study the expression of the endothelial and inducible isoforms of nitric oxide synthase (eNOS and iNOS, respectively) in human bladder carcinoma and schistosomal bladder disease, and to compare it with normal adult and fetal urothelium. Nitric oxide is thought to play a complex role in human carcinogenesis, but has only recently been investigated in bladder cancer. METHODS: Immunohistochemistry was performed on paraffin wax embedded sections of 33 human bladder carcinomas and five bladder carcinoma cell lines; in addition, seven schistosomal bladder cases and normal and fetal urothelium were investigated. In the cell lines enzymatic activity was examined by the NADPH diaphorase reaction. RESULTS: Immunoreactivity for eNOS was present in most cells of all 31 cases examined. Immunoreactivity for iNOS was less abundant and was seen in 23 of 25 cases. Similar findings were noted in schistosomal bladder cancer. In the normal bladder mucosa, eNOS immunoreactivity was found only in the superficial cell layer and iNOS was not expressed, whereas in the fetal urothelium immunoreactivity for both isoforms was seen in all cell layers. Enzymatic activity and immunoreactivity for eNOS and iNOS were evident in the five bladder carcinoma cell lines. CONCLUSIONS: It is possible that NOS plays a role in the differentiation of the transitional epithelium in fetal life, has a biological function in the adult bladder mucosa, and is involved in bladder carcinogenesis. eNOS and iNOS immunoreactivity do not differ in schistosomal and non-schistosomal bladder carcinoma, but resemble the pattern of expression typical of fetal urothelium. PMID- 11477141 TI - Application of laser capture microdissection and proteomics in colon cancer. AB - AIMS: Laser capture microdissection is a recent development that enables the isolation of specific cell types for subsequent molecular analysis. This study describes a method for obtaining proteome information from laser capture microdissected tissue using colon cancer as a model. METHODS: Laser capture microdissection was performed on toluidine blue stained frozen sections of colon cancer. Tumour cells were selectively microdissected. Conditions were established for solubilising proteins from laser microdissected samples and these proteins were separated by two dimensional gel electrophoresis. Individual protein spots were cut from the gel, characterised by mass spectrometry, and identified by database searching. These results were compared with protein expression patterns and mass spectroscopic data obtained from bulk tumour samples run in parallel. RESULTS: Proteins could be recovered from laser capture microdissected tissue in a form suitable for two dimensional gel electrophoresis. The solubilised proteins retained their expected electrophoretic mobility in two dimensional gels as compared with bulk samples, and mass spectrometric analysis was also unaffected. CONCLUSION: A method for performing two dimensional gel electrophoresis and mass spectrometry using laser capture microdissected tissue has been developed. PMID- 11477142 TI - Absence of CCND1 gene amplification in breast tumours of BRCA1 mutation carriers. AB - BACKGROUND/AIMS: It was recently reported that significantly fewer breast tumours of BRCA1 mutation carriers overexpressed cyclin D1 and HER2 protein than tumours of age matched breast cancer cases unselected for family history. This study aimed to examine the genetic basis of this reduction by determining the frequency of tumours within this cohort showing DNA amplification of these genes. METHODS: Paraffin wax embedded sections of breast tumours from BRCA1 mutation carriers and age, grade, histological type, and tumour size matched non-familial controls that had previously been stained for cyclin D1 and HER2 protein overexpression were analysed for CCND1 and HER2 gene amplification using fluorescence in situ hybridisation. RESULTS: CCND1 amplification was detected in none of the 30 tumours of the BRCA1 mutation carriers and in 19 of 74 tumours of the matched controls. Of those samples previously determined to overexpress the HER2 protein, HER2 amplification was detected in one of three tumours from BRCA1 mutation carriers and in 13 of 17 tumours of the age matched non-familial cases. CONCLUSION: None of the tumours of BRCA1 mutation carriers showed CCND1 amplification and only one tumour showed HER2 amplification. In contrast, a large proportion of cyclin D1 and HER2 overexpression in tumours of non-familial breast cancer cases could be accounted for by amplification of these genes. These data suggest that breast tumorigenesis in BRCA1 mutation carriers occurs by a molecular mechanism distinct from that of age matched non-familial cases. PMID- 11477143 TI - Epstein-Barr virus infection in paediatric liver transplant recipients: detection of the virus in post-transplant tonsillectomy specimens. AB - AIMS: Post-transplant lymphoproliferative disease (PTLD) is an important and serious complication in transplant patients. Recent studies have suggested that quantitative assessment of Epstein-Barr virus (EBV) infection in transplant patients might help to identify those at risk of developing PTLD. Therefore, tonsils from paediatric liver transplant recipients were studied for evidence of EBV infection. METHODS: Tonsils were studied by in situ hybridisation for the detection of the small EBV encoded nuclear RNAs (EBERs). The phenotype of EBV infected cells was determined by double labelling in situ hybridisation and immunohistochemistry. The expression of viral latent and lytic antigens was determined by immunohistochemistry. Tonsils from patients without known immune defects were studied as controls. RESULTS: Tonsils from transplant patients showed pronounced follicular hyperplasia and minor paracortical hyperplasia. In situ hybridisation revealed variable numbers of EBV infected B cells in the tonsils from transplant patients (range, 2-1000/0.5 cm(2); mean, 434/0.5 cm(2); median, 105/0.5 cm(2)). Lower numbers were detected in the control tonsils (range, 1-200/0.5 cm(2); mean, 47/0.5 cm(2); median, 9/0.5 cm(2)). The latent membrane protein 1 (LMP1) of EBV was not detected and there were only rare cells in two cases showing expression of the EBV encoded nuclear antigen 2 (EBNA2). There was no evidence of lytic infection. None of the patients developed PTLD within a follow up period of up to five years. CONCLUSIONS: These data indicate that tonsillar enlargement in paediatric liver transplant patients does not necessarily imply a diagnosis of PTLD. Furthermore, the presence of increased numbers of EBV infected cells in tonsils from liver transplant recipients by itself does not indicate an increased risk of developing PTLD. PMID- 11477144 TI - Chromosomal abnormalities in liver cell dysplasia detected by comparative genomic hybridisation. AB - BACKGROUND/AIM: The pathogenetic relation between liver cell dysplasia and hepatocellular carcinoma (HCC) is poorly understood. The aim of this study was to determine whether there is a genetic link between liver cell dysplasia and HCC that could support the role of dysplasia as a tumour precursor lesion. METHODS: Microdissection from paraffin wax embedded sections and degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) were combined to analyse chromosomal imbalances by comparative genomic hybridisation (CGH) in nine HCCs and nodules containing liver cell dysplasia and cirrhosis adjacent to the tumours. Seven cases of large cell changes (LCC) and three cases of small cell changes (SCC) were analysed. The genetic abnormalities detected in liver cell dysplasia were then compared with those present in the corresponding HCC. RESULTS: No abnormalities were detected in LCC and cirrhotic nodules, arguing against the preneoplasic nature of these cell foci. In contrast, a subset of chromosomal alterations present in HCCs was found in the adjacent SCC. CONCLUSIONS: These findings support the preneoplastic status of SCC in human hepatocarcinogenesis. PMID- 11477145 TI - Differential expression of the ccn3 (nov) proto-oncogene in human prostate cell lines and tissues. AB - AIMS: To investigate the expression of the human ccn3 (hccn3; nov) proto oncogene, a member of the CCN family of proteins, in prostate epithelial cells and prostate tissue samples. METHODS: Normal adult prostate luminal epithelial cells immortalised by SV40 large T (PNT1A and PNT1B), metastatic tumours (LNCaP, DU-145, and PC-3), and prostate tissue samples from patients with benign prostatic hyperplasia (BPH) and prostatic adenocarcinoma were used. hccn3 (nov) mRNA was measured by the reverse transcription polymerase chain reaction (RT-PCR) and hCCN3 (NOV) protein expression was determined by immunochemistry. RESULTS: hccn3 (nov) RNA values were higher in PC-3 cells than in the other prostate cell lines. The immortalised normal cell lines either did not express hccn3 (nov) RNA (PNT1B) or expressed very low amounts (PNT1A). BPH samples expressed variable amounts of hccn3 (nov) RNA. With the use of immunocytochemistry, all cell lines except PNT1A and PNT1B were shown to contain hCCN3 (NOV) protein. hCCN3 (NOV) was localised mainly in the epithelial compartment of BPH and adenocarcinoma samples, and there was evidence of luminal secretion. CONCLUSION: The overexpression of hccn3 (nov) RNA in cancer cell lines compared with other cell lines and its epithelial localisation in human prostate samples are consistent with a role for hCCN3 (NOV) in prostatic tumorigenesis. PMID- 11477146 TI - Role of thrombin receptors in the kidney: lessons from PAR1 knock-out mice. PMID- 11477147 TI - Vasopeptidase inhibitors--concepts and evidence. PMID- 11477148 TI - Guideline for the management of acute renal failure. PMID- 11477149 TI - The evidence base for the treatment of lupus nephritis in the new millennium. PMID- 11477150 TI - Advising dialysis patients to restrict fluid intake without restricting sodium intake is not based on evidence and is a waste of time. PMID- 11477151 TI - Vascular access: care and monitoring of function. PMID- 11477152 TI - Causes of death after renal transplantation. PMID- 11477153 TI - Ambulatory blood pressure monitoring: fancy gadgetry or clinically useful exercise? PMID- 11477154 TI - Acute dialysis quality initiative (ADQI). PMID- 11477155 TI - Gdansk bio-statistic and epidemiology course. PMID- 11477156 TI - Advanced glycation end products (AGEs)-induced expression of TGF-beta 1 is suppressed by a protease in the tubule cell line LLC-PK1. AB - BACKGROUND: Advanced glycation end products (AGEs) are assumed to play a key role in diabetic nephropathy (DN). Since little is known about their action in tubule cells, we investigated in LLC-PK1 cells: (i) whether AGE-bovine serum albumin (AGE-BSA) affects cell proliferation and expression of transforming growth factor beta (TGF-beta 1); and (ii) whether the AGE-induced effects can be modulated by trypsin due to interference with its binding proteins at the cell surface. METHODS: Arrested cells were exposed to vehicle (control), AGE-BSA (19--76 microM) and BSA (38 microM) in the presence or absence of trypsin (0.625--5.0 microg/ml) (2.5 microg/ml) for 24 h. We evaluated cell proliferation by cell count and by [(3)H]thymidine incorporation, TGF-beta 1 expression by reverse transcription-polymerase chain reaction (RT-PCR), and TGF-beta 1 protein by ELISA. In addition, cell accumulation of AGEs was studied by immunohistochemical staining of the AGE imidazolone. RESULTS: AGE-BSA inhibited [(3)H]thymidine incorporation, lowered cell number and increased cell protein content as well as TGF-beta 1 mRNA and protein as compared with control and BSA. Immunohistochemical staining revealed a marked intracellular accumulation of the AGE imidazolone. Co incubation of AGE-BSA with trypsin ameliorated the impaired thymidine incorporation, the decreased cell count and the enhanced cell protein content. TGF-beta 1 overexpression was normalized, while TGF-beta 1 protein declined insignificantly. Intracellular imidazolone accumulation was strikingly suppressed. CONCLUSIONS: In the tubule cell line LLC-PK1, AGE-BSA exerts an antiproliferative effect, most probably due to TGF-beta 1 overproduction. The co administration of trypsin abrogated this alteration, very likely as a result of an interaction with AGE-binding protein(s), which is supported by the decreased intracellular AGE accumulation. These findings may be the starting point for the development of specific proteolytic enzymes to interfere with the interaction between AGEs and their receptors/binding proteins. PMID- 11477157 TI - Amelioration of post-ischaemic renal injury by contralateral uninephrectomy: a role of endothelin-1. AB - BACKGROUND: Previous studies showed that unilateral renal damage is attenuated by prior contralateral uninephrectomy (Nx) in ischaemia-induced acute renal failure (ARF). Since renal ischaemia increases endothelin-1 (ET-1) production in the kidney, we examined whether the alteration of renal ET-1 content may contribute to the nephrectomy-induced attenuation of renal injury. METHODS: Ischaemic renal injury was provoked by 60-min left renal artery occlusion (RAO). Removal of the right kidney was performed just before RAO in the Nx group. Forty-eight hours after release of the clamp, renal ET-1 content was measured in both non nephrectomized and unilaterally nephrectomized rats. We also examined the effects of a selective ET(A) receptor (FR139317) and monoclonal ET antibody (AwETN40) on the RAO-induced changes in renal haemodynamics at 2 and 48 h after RAO respectively. RESULTS: The plasma concentration of ET-1 did not change in the two groups of ARF rats, but the cortical content of ET-1 increased to a lesser extent in Nx animals after ischaemia. Prior removal of the right kidney significantly facilitated the percentage recovery of left renal blood flow (RBF) during the first 2 h after release of the clamp. The percentage recovery of inulin clearance (Cin) by the kidney was also significantly better in Nx than sham-Nx rats at 48 h after RAO. Continuous administration of FR139317 (50 mg/kg/day) using osmotic minipumps for 3 days significantly attenuated exogenous ET-1-induced decrease in Cin and RBF. Infusion of FR139317 restored the decrease in RBF to control values during first 2 h in sham-Nx rats. However, FR139317 and AwETN40 did not ameliorate the RAO-induced decline of Cin in sham-Nx or Nx rats at 2 and 48 h after ischaemia respectively. CONCLUSIONS: Contralateral uninephrectomy prior to ischaemia-induced ARF attenuated the increase in cortical ET-1 content and subsequent renal response to ischaemic injury. This beneficial effect of unilateral nephrectomy, however, was not mediated through well-preserved RBF due to reduced intrarenal ET-1 action. PMID- 11477158 TI - Effect of mycophenolate mofetil on nitric oxide production and inducible nitric oxide synthase gene expression during renal ischaemia-reperfusion injury. AB - BACKGROUND: Recent animal data suggest that inducible nitric oxide synthase (iNOS) derived nitric oxide (NO) plays an important role in the pathogenesis of renal ischaemia-reperfusion injury (IRI) and that inhibition of iNOS ameliorates IRI. Mycophenolate mofetil (MMF), a lymphocyte selective anti-proliferative agent, has been shown to inhibit NO production in vitro. The aim of this study is to evaluate the effect of MMF on NO production and iNOS gene expression in vivo during renal IRI. METHODS: Renal IRI was induced by clamping the left renal pedicle of male BALB/c mice for 30 min, followed by 15 min of reperfusion. The mice received placebo or MMF at 40, 80 or 120 mg/kg/day by oral gavage for 5 days before the operation. Sham-operated mice served as the operation control. The amount of NO produced and the level of iNOS gene expression in the kidney tissue during IRI was assessed by spin trapping electron paramagnetic resonance (EPR) spectroscopy and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) respectively. RESULTS: In the sham-operated kidneys, only low levels of NO and iNOS mRNA were detected. In mice with renal IRI, the amount of NO detected was significantly increased, which was reduced in a dose dependent fashion by pre-treatment with MMF. Pre-treatment with MMF also substantially reduced iNOS gene expression in the kidney tissue. CONCLUSIONS: We conclude that pre-treatment with MMF inhibits the production of NO and the induction of iNOS gene expression in the kidney during IRI. These results suggest that MMF might have the potential to ameliorate renal IRI. PMID- 11477159 TI - alpha-MSH decreases apoptosis in ischaemic acute renal failure in rats: possible mechanism of this beneficial effect. AB - BACKGROUND: Apoptosis frequently occurs in acute renal injury but the molecular mechanisms responsible for this distinct form of cell death are largely unknown. Fas belongs to the tumour necrosis factor (TNF)/nerve growth factor superfamily and engagement by Fas ligand induces apoptosis in various epithelial cells. To investigate the role of apoptosis and associated mechanisms, we examined the occurrence of apoptosis and Fas and Fas ligand expression, and the therapeutic effect of alpha-melanocyte-stimulating hormone (alpha-MSH), a potent anti inflammatory cytokine in an ischaemic acute renal failure (ARF) rat model. We also examined neutrophil infiltration together with intercellular adhesion molecule-1 (ICAM-1) expression because of their possible involvement in apoptosis due to their ability to release various inflammatory cytokines and reactive oxygen species. METHODS: After unilateral nephrectomy in female Sprague-Dawley rats, the renal artery of the contralateral kidney was clamped for 40 min and reperfused. alpha-MSH or vehicle was injected intraperitoneally immediately after reperfusion and at 1, 6, or 24 h after reperfusion. The expression of Fas and Fas ligand was studied by western blot analysis and semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and neutrophil infiltration by naphthol AS-D chloracetate staining. The degree of apoptosis, neutrophil infiltration, and Fas and Fas ligand, and ICAM-1 expression, as well as biochemical and histological data were compared between the alpha-MSH and the vehicle-treated groups. RESULTS: Intraperitoneally administered alpha-MSH significantly reduced renal injury, measured by blood urea nitrogen (BUN) and creatinine and by the degree of tubular necrosis (109.6+/-7.1/54.7+/-3.1 mg/dl for BUN, and 1.6+/-0.2/1.03+/-0.06 mg/dl for creatinine 24 h after ischaemia) (5.4+/-0.8/2.6+/-0.3 for injury score 24 h after ischaemia). Ischaemia caused an increase in Fas and Fas ligand expression and was accompanied by morphological evidence of apoptosis. alpha-MSH significantly reduced the degree of apoptosis, as well as Fas and Fas ligand expression (mean apoptotic cell number, 41.7+/-3.5/14.2+/-2.2 per x200 field at 24 h after ischaemia. Fas protein expression: sham, 1409+/-159 DI (densitometric index); vehicle/alpha-MSH, 2818+/-635/1306+/-321 DI at 24 h and 5542+/-799/2867+/ 455 DI at 72 h after ischaemia. Fas ligand protein expression: sham, 1221+/-181 DI; vehicle/alpha-MSH, 2590+/-85/1279+/-169 DI at 4 h, 4376+/-268/2432+/-369 DI at 24 h and 5200+/-648/2253.7+/-1104 DI at 72 h after ischaemia). Neutrophil infiltration and ICAM-1 expression were also significantly reduced in alpha-MSH group (neutrophil infiltration: vehicle/ alpha-MSH, 5.05+/-1.8/1.59+/-0.4) (ICAM 1 expression, vehicle/alpha-MSH 0.46+/-0.21/0.29+/-0.19). CONCLUSION: These results suggest that apoptosis clearly contributes to tubular cell loss in ischaemia/reperfusion (I/R) injury possibly by neutrophil-mediated pathways or an increase in Fas-Fas ligand expression. The observed beneficial effect of alpha MSH could be related to these mechanisms. PMID- 11477160 TI - Haemodynamic effects of valsartan in acute renal ischaemia/reperfusion injury. AB - BACKGROUND: Acute deterioration of renal function is an important side-effect of angiotensin-converting enzyme (ACE) inhibitors, especially if accompanied by other nephrotoxic events. Angiotensin II receptor(1) blockers (ARB) are thought to have fewer side-effects on renal perfusion and function. We examined the effects of valsartan (VAL) on kidney function as well as the contribution of the nitric oxide (NO) system in a rat model of ischaemic acute renal failure (ARF). METHODS: ARF was induced by 40 min of clamping of both renal arteries in female Sprague-Dawley rats. Renal haemodynamic and tubular parameters were determined during post-ischaemic infusion of vehicle, VAL, VAL and the NO-synthase substrate L-arginine, and VAL together with inhibition of NO synthases (NOS) by L-NMMA. RESULTS: Clamping induced acute renal failure with marked decreases in glomerular filtration rate (GFR) and renal plasma flow (RPF) accompanied by a rise in renal vascular resistance (RVR) and fractional sodium excretion. Valsartan caused a slight but significant improvement of GFR and RPF without full recovery of renal function and caused a lowering of RVR and tubular sodium loss. L-arginine-co administration had no additive beneficial effect. Valsartan-induced changes were not significantly depressed by unspecific inhibition of NOS. CONCLUSIONS: Inhibition of the angiotensin II-receptor(1) diminishes the deleterious effects of ischaemia and reperfusion on glomerular function and on the renal microcirculation. An involvement of the NO system could not be demonstrated. PMID- 11477161 TI - Conjugated dienes: a critical trait of lipoprotein oxidizability in renal fibrosis. AB - BACKGROUND: We assessed whether a differential oxidizability of apolipoprotein B (apo B)-containing lipoproteins (LDL and VLDL) may explain the oxidative stress that we had observed at the onset of renal fibrosis in Zucker obese (ZO) rats (Nephrol Dial Transplant 2000, 15: 467--476). METHODS: Ex vivo copper-induced oxidation of lipoproteins was performed in 1-, 3-, and 9-month-old ZO and age matched lean (ZL) rats. LDL/VLDL oxidizability was determined by spectrophotometry at 234 nm by monitoring the formation of conjugated diene hydroperoxides. RESULTS: A significant increase in lag time (reflecting the resistance to oxidation) was observed in ZO rats at 3 months while the maximal diene production (reflecting the amount of hydroperoxides formed during oxidation) was higher in ZO than in ZL rats as early as 1 month. Lipoproteins were larger in ZO than in ZL rats, as shown by their core to surface component ratio. Furthermore, ZO lipoproteins had increased vitamin E and polyunsaturated fatty acid (PUFA) content, with no change in vitamin E/PUFA ratio. CONCLUSIONS: Rather than oxidizability of apo B-containing lipoproteins, the ability of these molecules to produce high levels of conjugated dienes, which can act as toxic tissue messengers, appears to be a critical trait in the development of renal fibrosis in this rat model of obesity and renal fibrosis. PMID- 11477162 TI - Effect of chronic renal failure on the expression and function of rat intestinal P-glycoprotein in drug excretion. AB - BACKGROUND: In chronic renal failure, the renal excretion of certain drugs is dramatically reduced. To determine whether other routes of drug elimination, such as secretion through the intestinal barrier by intestinal P-glycoprotein can be altered, we compared P-glycoprotein activity, P-glycoprotein protein content, and P-glycoprotein mRNA levels in intestine of control and chronic renal failure rats. METHODS: Chronic renal failure was surgically induced in rats by partial (7/8) nephrectomy. After 5 weeks, intestinal transport of rhodamine 123, a P glycoprotein substrate, was carried out using an in vitro model of everted gut sacs. P-glycoprotein protein content was quantified by enzyme-linked immunosorbent assay and P-glycoprotein mRNA expression was evaluated by semi quantitative reverse transcriptase polymerase chain reaction. RESULTS: A decrease of intestinal rhodamine 123 transport was observed in chronic renal failure rats, pointing to an inhibition of P-glycoprotein activity. Transport was inhibited in both sham-operated rats and rats with chronic renal failure by verapamil and cyclosporin A, but relative inhibition vs baseline was less marked in chronic renal failure than in sham-operated rats. In contrast, no significant differences in levels of P-glycoprotein protein or mRNA were observed between the two groups. CONCLUSIONS: Intestinal secretion of rhodamine 123 is mainly mediated by P glycoprotein. It was reduced in rats with chronic renal failure, reflecting reduced intestinal drug elimination via a decrease in P-glycoprotein transport activity rather than via protein underexpression. PMID- 11477163 TI - Assessment of haematuria: automated urine flowmetry vs microscopy. AB - BACKGROUND: Microscopy of the urine sediment may be a useful method in the distinction between a glomerular and a non-glomerular source of urinary bleeding. However, microscopic techniques are time consuming and hampered by inter-observer variations. In the present study we have therefore compared bright-field microscopy with automated urine flowmetry (Sysmex UF-100), examining their ability to differentiate between glomerular and non-glomerular haematuria. METHODS: Fresh urine samples were obtained from 112 patients with a well-defined, single cause of a positive dipstick test. Their urine specimens were examined within 4 h in a blinded manner. Of them, 79 specimens had a positive dipstick for blood and thus could be evaluated for haematuria. RESULTS: The Sysmex UF-100 had a sensitivity and specificity of 0.83 and 0.94 respectively in detecting non glomerular bleeding. The positive and negative predictive values were 0.95 and 0.78 respectively. The corresponding values of microscopy were 0.79 and 0.90 respectively, and 0.93 and 0.74 respectively. CONCLUSIONS: Automated flowmetry can be used in the distinction between glomerular and non-glomerular haematuria. PMID- 11477164 TI - Diagnosis of nut-cracker phenomenon using renal Doppler ultrasound in orthostatic proteinuria. AB - BACKGROUND: We evaluated the efficacy of non-invasive renal Doppler ultrasound (US) to detect the nut-cracker phenomenon (NCP) and we studied the prevalence of NCP in children with orthostatic proteinuria. METHODS: Among a total 66 cases of orthostatic proteinuria, 39 cases of NCP were found, with 27 cases being detected in a normal control group. Using Doppler US, the anteroposterior (AP) diameter and peak velocity (PV) of the left renal vein (LRV) were measured at the hilar and aortomesenteric portion. We calculated the ratio of AP and PV diameters between the two portions. The parameters were analysed using Student's t-test. RESULTS: The AP diameters and the ratio in the hilar and narrow portions were all significantly different between the two groups (P<0.01). The PV in the narrow portion and the ratio of PV were significantly different (P<0.01), but the PV in the hilar portion was not statistically different between the two groups (P>0.05). If the diagnostic criteria for NCP was that the ratio of PV was more than 5, then 22 subjects (56.4%) in the orthostatic proteinuria group and none in the control group could be diagnosed as NCP. If, however, the cut-off values for the diagnosis of NCP were set at the mean+/-2 SD of the ratio (PV ratio 3.98 and size ratio 4.16), then the orthostatic proteinuria group showed abnormal AP diameter in 25 (64.1%), peak velocity in 28 (71.8%), and both in 21 patients (53.8%), and the control group showed an abnormal AP diameter in one subject (3.7%). CONCLUSIONS: NCP may be one of the leading causes of orthostatic proteinuria, and non-invasive renal Doppler US may be a useful diagnostic tool in the screening of NCP. In the future, the diagnostic criteria of NCP must be redefined in children. PMID- 11477165 TI - Oxidant stress in nephrotic syndrome: comparison of F(2)-isoprostanes and plasma antioxidant potential. AB - BACKGROUND: The nephrotic syndrome (NS) is associated with an increased risk of coronary heart disease. Increased oxidant stress may contribute to this by means of hyperlipidaemia and/or hypoalbuminaemia. In this study we assessed the contributory role of oxidant stress, as measured by F(2)-isoprostanes and plasma oxygen radical absorbance capacity (ORAC), in subjects with NS. METHODS: We studied 14 subjects with NS and 17 age- and sex-matched healthy non-proteinuric controls. Measurement of plasma and urinary F(2)-isoprostanes was carried out using a combination of silica and reverse-phase cartridges, high-performance liquid chromatography, and gas chromatography mass spectrometry using electron capture negative ionization. The plasma ORAC assay measured the decrease in fluorescence of phycoerythrin added to plasma in the presence of a free-radical generator. The ORAC value (microM) was calculated as the ratio of the area under the fluorescence decay curve for plasma to the area under the fluorescence decay curve for a Trolox standard. RESULTS: Plasma ORAC was significantly lower in NS patients compared with controls: mean (standard error) NS patients 3306 microM (286); controls 4882 microM (496), P=0.011. In univariate linear regression analysis, plasma albumin was significantly positively correlated with plasma ORAC (r=0.40, P=0.03). Plasma and urinary F(2)-isoprostanes did not differ significantly between NS and control groups. CONCLUSIONS: This study demonstrates that in the NS there is decreased free-radical trapping capacity of plasma that is inversely correlated with hypoalbuminaemia, but no increase in plasma and urinary F(2)-isoprostanes. Decreased total plasma antioxidant potential in combination with hyperlipidaemia may contribute to the increased risk of cardiovascular disease seen in NS. PMID- 11477166 TI - ANCA titres, even of IgG subclasses, and soluble CD14 fail to predict relapses in patients with ANCA-associated vasculitis. AB - BACKGROUND: Antineutrophil cytoplasmic autoantibodies (ANCA) are presumed to reflect disease-activity and to be useful for guidance of immunosuppressive therapy of ANCA-associated systemic vasculitis (AASV), but with respect to conventional ANCA assays this is controversial. ANCA titres, measured in the IgG3 subclass and modern capture ELISAs, have been said to be superior predictors of relapses of AASV. METHODS: In this retrospective study serial measurements of ANCA parameters and soluble CD14 (sCD14) were performed in 169 consecutive sera over a median of 21 months in 18 patients with AASV and related to disease activity, assessed by Birmingham Vasculitis Activity Score (BVAS) for new or deteriorated (BVAS1), and for chronic disease activity (BVAS2). Fourteen patients had Wegener's granulomatosis (WG) and were C-ANCA positive with Pr 3-antibodies and four patients had microscopic polyangiitis (MPA) with P-ANCA and MPO antibodies. In WG patients ANCA by IIF, Pr 3-ELISA for IgG, IgG1, IgG3, IgG4 and sCD14 were measured, as well as capture ELISA for Pr 3, and in MPA patients ANCA by IIF, MPO-ELISA for IgG and IgG1, IgG3, IgG4, and sCD14 respectively. In eight patients, data collection started at diagnosis, in 10 patients at remission. RESULTS: The parameters predicted neither the nine major relapses (increase of immunosuppression necessary), nor the 26 minor relapses (increase of BVAS1>2) with sufficient sensitivity (>80%) or specificity (> 90%90%), and they also failed to predict relapses within the following 2 months. ANCA-IgG3 and capture ELISA for Pr 3 were not advantageous for prediction of relapses (sensitivity 0.45 and 0.19 respectively), and sCD14 remained elevated in all samples irrespective of disease activity. CONCLUSIONS: There is no rationale for serial measurements of ANCA in AASV. For changes of therapy, the ANCA parameters should only be used in conjunction with clinical information. PMID- 11477167 TI - Local macrophage proliferation correlates with increased renal M-CSF expression in human glomerulonephritis. AB - BACKGROUND: Macrophage accumulation is a prominent feature in many forms of glomerulonephritis. Local proliferation of macrophages within the kidney has been described in human and experimental glomerulonephritis and may have an important role in augmenting the inflammatory response. The current study examined the relationship between local macrophage proliferation and renal expression of macrophage colony-stimulating factor (M-CSF). METHODS: A total of 118 renal biopsies of patients with a wide range of glomerulonephridities were examined for M-CSF protein and macrophage proliferation (KP1+PCNA+cells) by single and double immunohistochemistry staining, respectively. RESULTS: Biopsies of thin membrane disease (TMD) with histologically normal kidney showed M-CSF protein expression by 33% of cortical tubules, while glomerular M-CSF expression was limited to resident macrophages and some podocytes. Glomerular M-CSF expression increased significantly in proliferative forms of glomerulonephritis, with M-CSF staining of infiltrating macrophages, podocytes and some mesangial cells. Segmental areas of strong M-CSF expression, particularly in crescents, co-localized with KP1+PCNA+ proliferating macrophages. There was also an increase in tubular M-CSF expression in most types of glomerulonephritis. Tubular M-CSF staining was strongest in areas of tubular damage and co-localized with KP1+ macrophages, including KP1+PCNA+ proliferating macrophages. Many interstitial macrophages and alpha-smooth muscle actin-positive myofibroblasts showed strong M-CSF staining. Statistical analysis showed a highly significant correlation between M-CSF expression and local macrophage proliferation in both the glomerulus and tubulointerstitium. Glomerular and tubular M-CSF expression gave a significant correlation with renal dysfunction. CONCLUSIONS: Glomerular and tubulointerstitial M-CSF expression is up-regulated in human glomerulonephritis, being most prominent in proliferative forms of disease. This correlated with local macrophage proliferation, suggesting that increased renal M-CSF production plays an important role in regulating local macrophage proliferation in human glomerulonephritis. PMID- 11477168 TI - Autosomal recessive polycystic kidney disease in adulthood. AB - BACKGROUND: Renal cysts arising from collecting ducts, congenital hepatic fibrosis, and recessive inheritance characterize autosomal recessive polycystic kidney disease (ARPKD). The disorder usually manifests in infancy, with a high mortality rate in the first year of life. For the patients who survive the neonatal period, the probability of being alive at 15 years of age ranges from 50 to 80%, with 56--67% of them not requiring renal replacement therapy at that stage. Some develop portal hypertension. Long-term outcome of adults escaping renal insufficiency above age 18 is largely unknown. METHOD: In consecutive patients with ARPKD and autonomous renal function at age 18, clinical course of kidney and liver disease in adulthood and status at last follow-up were evaluated. Progression of renal insufficiency was assessed by the rate of decline of creatinine clearance, according to Schwartz's formula before age 18 and Cockcroft and Gault formula thereafter. Severity of liver involvement was estimated by imaging techniques, liver function tests, and endoscopy. RESULTS: Sixteen patients from 15 families were included. ARPKD was diagnosed between 1 day and 13 years of age. From diagnosis, mean follow-up period lasted 24+/-9 years. Before age 18, nine patients (56%) were hypertensive, nine (56%) had renal failure, and four (25%) had portal hypertension. Beyond age 18, no additional patient became hypertensive, and another five developed progressive renal insufficiency; altogether, the mean yearly decline of creatinine clearance was 2.9+/-1.6 ml/min. Portal hypertension was recognized in two additional patients. Four patients experienced gastro-oesophageal bleeding, while recurrent cholangitis or cholangiocarcinoma developed in one case each. At the end of follow-up, 15/16 patients (94%) were alive at a mean age of 27 (18--55) years. Two patients had a normal renal function, 11 had chronic renal insufficiency, one was on regular dialysis, and two had functioning kidney grafts. Four patients had required a porto-systemic shunt. CONCLUSIONS: A subset of ARPKD patients with autonomous renal function at age 18 experiences slowly progressive renal insufficiency. With prolonged renal survival, complications related to portal hypertension are not rare, requiring careful surveillance and appropriate management. PMID- 11477169 TI - Diabetic microvascular complications are not associated with two polymorphisms in the GLUT-1 and PC-1 genes regulating glucose metabolism in Caucasian type 1 diabetic patients. AB - BACKGROUND: An XbaI polymorphism in the gene encoding the glucose transporter, GLUT-1, is associated with development of diabetic nephropathy in Chinese type 2 diabetic patients. In addition, an amino acid variant (K121Q) in the gene encoding the glycoprotein plasma cell differentiating antigen (PC-1), a specific inhibitor of insulin receptor signalling, has been reported to predict a faster progression of nephropathy in Italian and British type 1 diabetic patients. METHODS: The XbaI and K121Q polymorphisms were determined by PCR-RFLP in Danish type 1 diabetic patients with nephropathy (122 men/77 women, age 40.9+/-9.6 years, diabetes duration 27+/-8 years) and type 1 diabetic patients with persistent normoalbuminuria (118 men/74 women, age 42.7+/-10.2 years, diabetes duration 26+/-9 years). Proliferative retinopathy was present in 156 patients (40%), while 67 patients (17%) had no diabetic retinopathy. RESULTS: There were no differences in the frequency of GLUT-1 XbaI genotypes between type 1 diabetic patients with diabetic nephropathy and type 1 diabetic patients with normoalbuminuria: 72 (41%)/87 (50%)/16 (9%) vs 94 (49%)/74 (39%)/24 (13%) had GLUT-1 XbaI +/+, +/- or -/- genotype respectively (NS). The frequency of PC-1 KK, KQ and QQ genotypes were 141 (71%)/52 (26%)/6 (3%) vs 138 (73%)/45 (24%)/7 (4%) in patients respectively with and without nephropathy (NS). Neither were associations between the investigated polymorphisms and simplex or proliferative retinopathy revealed. CONCLUSIONS: Neither the PC-1 K121Q nor the GLUT-1 XbaI polymorphism contribute to the genetic susceptibility of diabetic microvascular complications in Danish type 1 diabetic patients. PMID- 11477170 TI - Autonomic insufficiency as a factor contributing to dialysis-induced hypotension. AB - BACKGROUND: Autonomic insufficiency is considered a factor that contributes to dialysis-induced hypotension (DIH). However, the relationship between the two conditions has not been fully elucidated. METHODS: We investigated 44 haemodialysis patients using [(123)I]-meta-iodobenzylguanidine (MIBG) scintigraphy and power-spectral analysis (PSA) of heart rate variability. The patients were divided into four groups: a diabetic group with DIH, a diabetic group without DIH, a non-diabetic group with DIH, and a non-diabetic group without DIH. In these groups the heart to mediastinum average count rate (H/M), MIBG washout rate, and low- and high-frequency components of PSA were compared. RESULTS: From the [(123)I]-MIBG scintigraphy, for both early and delayed images, H/M of the groups with DIH were lower than in groups without DIH, in both diabetics and non-diabetics (P<0.05). For the early images, H/M of the diabetic groups were lower than in the non-diabetic groups, in the groups both with and without DIH (P<0.01). For the delayed images, H/M of the diabetic group was lower than in the non-diabetic group, in the groups with DIH (P<0.05). The MIBG washout rate was the highest in the diabetic group with DIH (P<0.05 vs diabetic and non diabetic groups without DIH). The PSA of heart rate variability showed a good discrimination of the low-frequency component between the non-diabetic patients with and without DIH (P<0.05). Mean ultrafiltration volume and its rate were not different among the four groups. CONCLUSION: Autonomic insufficiency is more severe in patients with DIH than in those without, and its degree may be enhanced in diabetic patients. For the management of DIH, special care should be addressed not only to dry weight but also to autonomic insufficiency. PMID- 11477171 TI - Acute effects of haemodialysis on endothelial function and large artery elasticity. AB - BACKGROUND: Disturbances of functional properties of large arteries contribute to increased cardiovascular morbidity and mortality in patients with end-stage renal disease. However, it is not clear whether haemodialysis per se acutely affects mechanical vessel wall properties or endothelial function. METHODS: Twenty-five chronic haemodialysis patients (mean+/-standard error of the mean (SEM): age 52+/ 5 years; time on dialysis 63+/-7 months; blood pressure 132+/-4/72+/-2 mmHg) were studied before and immediately after a haemodialysis (HD) session using a polysulphone dialyser (ultrafiltration 1460+/-54 ml), as well as on the following day. Blood pressure was measured with an automatic sphygmomanometer and applanation tonometry. End-diastolic diameter and distension of the brachial and carotid arteries were measured by Doppler frequency analysis of vessel wall movements in M-mode using a multigate pulsed Doppler system and aortic pulse wave velocity (PWV) by an automatic device (Complior). Endothelial function was determined as brachial artery flow-mediated dilation (FMD) and compared with endothelium-independent nitroglycerine-induced dilation (NMD). RESULTS: FMD was 7.9+/-1.8% in patients before HD and did not change significantly after HD or in the dialysis-free intervall (6.7+/-2.1 and 7.1+/-2.0%, respectively; NS). The same was true for NMD and PWV (12.6+/-0.8 m/s before HD, 12.8+/-0.8 m/s after HD, and 11.9+/-0.7 m/s on the HD-free day). Carotid distensibility coefficients decreased significantly during HD (from 18.1+/-1.9 x 10(-3)/kPa to 16.7+/-2.2 x 10(-3)/kPa, P<0.05) and increased again on the HD-free day (19.8+/-2.4 x 10( 3)/kPa). However, when corrected for blood pressure by tonometry, isobaric carotid distensibility did not change significantly. Brachial artery distensibility also did not show significant acute changes. CONCLUSIONS: Haemodialysis per se did not have a significant effect on endothelial function or large artery mechanical vessel wall properties in patients on maintenance dialysis therapy. PMID- 11477172 TI - Risk of death and liver cirrhosis in anti-HCV-positive long-term haemodialysis patients. AB - BACKGROUND: Hepatitis C virus (HCV) infection is the most common cause of chronic liver disease in haemodialysis patients. The aim of this study was to assess the impact of HCV infection on patient survival in a cohort of long-term haemodialysis patients and to evaluate the percentage of anti-HCV-positive patients that evolve to liver cirrhosis. METHODS: In 1992, 175 patients who had been on intermittent haemodialysis therapy for at least 6 months were included in the study (57 anti-HCV-positive and 118 anti-HCV-negative patients). Evaluation of patient outcome included date and cause of death, kidney transplantation, and the diagnosis of liver cirrhosis. Patient survival was estimated by the Kaplan Meier method and compared by the log-rank test. The Cox proportional hazards model was used to estimate the risk of death among dialysis patients who were anti-HCV positive. Other prognostic variables studied included age, gender, diabetes mellitus as cause of end-stage renal disease (ESRD), history of previous transplant, transplantation during follow-up, and time on haemodialysis treatment. The diagnosis of liver cirrhosis was made based on clinical and/or histological criteria. RESULTS: Eight-year patient survival in anti-HCV-positive subjects was lower (32%) than in anti-HCV-negative patients (52%) (log-rank, P=0.03). Four variables were found to be independent prognostic factors in patient survival: age (relative risk (RR) 1.04); diabetes as cause of ESRD (RR 3.6); transplantation during follow-up (RR 0.66) and presence of HCV antibodies (RR 1.62). The causes of death did not differ significantly between groups, except that four anti-HCV-positive patients died from liver disease. Ten (17.5%) of the 57 anti-HCV-positive patients were diagnosed to have liver cirrhosis at a median of 10 years after renal replacement therapy initiation and a median of 7 years after the first ALT level increase. CONCLUSION: In conclusion, our study shows an increased risk of death among long-term haemodialysis patients infected with HCV compared with non-infected patients. This might be partly explained by the high proportion of these patients that evolve to liver cirrhosis. PMID- 11477173 TI - Is the loss of health-related quality of life during renal replacement therapy lower in elderly patients than in younger patients? AB - BACKGROUND: Previous studies have reported that elderly (aged 65 years or over) end-stage renal disease (ESRD) patients have poorer health-related quality of life (HRQOL) than both younger patients and healthy subjects of the same age. The aim of present study was to evaluate the effect of ESRD and its treatment on the HRQOL, and to determine the effects of age and gender. METHODS: A cross-sectional multicentric study was carried out with 485 haemodialysis and renal-transplant patients, using the SF-36 Health Survey to evaluate their HRQOL. SF-36 scores were standardized by age and gender using Spanish normative data. Karnofsky scale score (KS), socio-demographic, and clinical data were also collected. RESULTS: In renal-replacement therapy (RRT), chronic haemodialysis, and renal-transplant patients, SF-36 standardized scores of elderly patients were higher than in younger patients. Therefore the reduction in HRQOL of elderly patients, in relationship with that of the general population of the same age and gender, was lower than in younger patients. In the case of renal-transplant patients, standardized scores in elderly patients were higher than in the general population for all parameters. CONCLUSIONS: Using standardized scores, elderly patients on renal replacement therapy (haemodialysis and kidney transplant) had relatively better HRQOL than younger patients, and in the case of transplant patients, they had even better HRQOL than in the general population of the same age and gender. PMID- 11477174 TI - Outcome in patients with end-stage renal disease following heart or heart--lung transplantation receiving peritoneal dialysis. AB - BACKGROUND: End-stage renal disease (ESRD) complicates 5--10% of heart and heart- lung transplant patients. We report our experience of peritoneal dialysis (PD) in 17 such patients. METHODS: Between March 1995 and February 1999, 13 heart transplant and four heart--lung transplant patients (11 male, 6 female) joined our PD programme (10 continuous ambulatory PD, seven automated PD). Median time from heart or heart--lung transplantation to ESRD was 9 years (range 1--13 years), and median age at introduction of renal replacement therapy was 51 years (range 23--66 years). The frequency of exit-site infections, peritonitis, and PD survival (including technique failure and death) in the transplant group (TxP) was calculated retrospectively. These were compared with two contemporary control groups: PD patients immunosuppressed for other indications (ISP, n=19) and, all other patients recruited onto the PD programme (NISP, n=132). RESULTS: Median follow-up was 10 months (range 2--27 months) for TxP, 7 months (range 2--29 months) for ISP, and 14 months (range 1--48 months) for NISP groups. The frequency of exit-site infections was similar in each group: 1 in 26 months for TxP; 1 in 30 months for ISP, and 1 in 27 months for NISP (P=NS). The frequency of peritonitis was greater in the TxP group at 1 in 15 months, compared with 1 in 20 months for ISP and 1 in 29 months for NISP (TxP vs NISP, P<0.05). PD failure following infection was 23.5% for TxP, 10.5% for ISP, and 12.9% for NISP. Actuarial PD survival at 24 months was only 25.2% in the TxP group compared with 79% in the NISP group. There were no deaths related to immediate complications of PD. CONCLUSIONS: Increased risk of PD peritonitis and reduced PD survival is reported in this cohort of 17 heart and heart--lung recipients with ESRD. Nevertheless, for patients with severely impaired cardiac function, PD may still offer therapeutic advantage. PMID- 11477175 TI - Outcome of cadaver kidney transplantation in 23 patients with type 2 diabetes mellitus. AB - BACKGROUND: Type 2 diabetes mellitus (DM) is a growing cause of end-stage renal failure worldwide. Yet, only a minority of type 2 diabetics are considered today for kidney transplantation (KT). The scarcity of data on the outcome of such patients after KT prompted us to review our experience. METHODS: Between 1 January 1983 and 30 June 1996, 23 patients with type 2 DM received a first cadaver KT at a mean age of 57+/-9 (41-73) years, after a dialysis period ranging from 5 to 72 (mean 25+/-18) months. Only nine patients had a history of coronary and/or peripheral vascular disease before KT. All were given cyclosporin- or tacrolimus-based immunosuppression. Post-KT follow-up ranged from 4 to 181 (mean 70+/-38) months. Outcome analysis focused on the impact of cardiovascular complications. RESULTS: Patient survival at 1, 5 and 8 years was 91, 83 and 76% respectively. Death was due to infection in three patients and to a cardiovascular event in two. The actuarial risk of coronary, cerebrovascular, peripheral vascular, and any cardiovascular event after KT was 14, 13, 9 and 30% at 1 year, 20, 13, 50 and 58% at 5 years, and 20, 46, 66 and 72% at 8 years respectively. Post-KT hospital readmissions averaged 10 days/patient-year and were mostly related to the management of peripheral vascular disease. CONCLUSION: KT is an excellent therapeutic option for selected patients with type 2 DM. Peripheral vascular disease is the leading cause of morbidity following KT. KT should be considered in type 2 diabetics with a low/medium cardiovascular risk. PMID- 11477176 TI - A possible role of thrombin-activatable fibrinolysis inhibitor in disturbances of fibrinolytic system in renal transplant recipients. AB - BACKGROUND: Cardiovascular disease (CVD) is a major cause of death in renal transplant recipients (RTR). Suppression of fibrinolysis plays a role in the progression of atherosclerosis. Accelerated progression of atherosclerosis and fibrinolytic system suppression has been observed in RTR. Despite many years of intensive research, the reason for impaired fibrinolysis in this patient population is not fully understood. Thrombin-activatable fibrinolysis inhibitor (TAFI) is a recently discovered glycoprotein combining coagulation and fibrinolysis. This study was conducted to evaluate concentrations of TAFI, markers of thrombin generation, endothelial injury, and some standard laboratory parameters in RTR receiving triple immunosuppressive drug regimen. METHODS: The study was performed in 29 stable, non-diabetic kidney transplant recipients treated with cyclosporin A, azathioprine, and prednisone and in 18 age- and sex matched healthy volunteers. Soluble thrombomodulin (sTM), prothrombin fragments F1+2 (F1+2), thrombin--antithrombin complexes (TAT), plasmin--antiplasmin complexes (PAP), and TAFI were measured with commercially available kits. RESULTS: The RTR group had significantly higher plasma levels of TAT, F1+2, sTM and TAFI than the healthy volunteers. There were no differences in PAP concentrations between the two groups. Plasma sTM correlated inversely with creatinine clearance, body mass index, haemoglobin, and albumin. Plasma TAT level was positively associated with total cholesterol. TAFI antigen influenced negatively PAP antigen concentration. CONCLUSIONS: On the basis of our research, we concluded that elevated circulating TAFI antigen might be a new link in the pathogenesis of impaired fibrinolysis in RTR, and thus atherosclerosis progression. In the patient group there is also evidence of endothelial injury, followed by secondary activation of the coagulation cascade. Hypercholesterolaemia in RTR is associated with enhanced thrombin activity. PMID- 11477177 TI - Primary hyperoxaluria type 2 without L-glycericaciduria: is the disease under diagnosed? PMID- 11477178 TI - Myoglobinuric acute renal failure following cardioversion in a boxfish poisoning patient. PMID- 11477179 TI - Mycophenolate mofetil causing deep venous thrombosis in a renal transplant patient with factor V Leiden. PMID- 11477180 TI - Late allograft loss due to recurrence of p-ANCA-associated systemic vasculitis in a patient with relapsing polychondritis. PMID- 11477181 TI - Severe hyponatraemia due to hypothalamic--pituitary adrenal insufficiency. PMID- 11477182 TI - Acute renal infarction in a woman with slight asthma. PMID- 11477183 TI - Acute renal failure in a single kidney with previous obstruction. PMID- 11477184 TI - Purple-blue subcutaneous nodules after renal transplantation: not always Kaposi sarcoma. PMID- 11477185 TI - Timing of sildenafil therapy in dialysis patients. PMID- 11477186 TI - Homocysteine and nephrotic syndrome. PMID- 11477187 TI - Outcome of renal replacement therapy in the very elderly. PMID- 11477188 TI - Lymphomas and IgA nephropathy. PMID- 11477189 TI - Allergic reactions to oral cyclophosphamide therapy in immunologically-mediated renal disease. PMID- 11477190 TI - Laser therapy in patient with intractable haemorrhage due to the bladder involvement of systemic amyloidosis. PMID- 11477191 TI - Thrombotic microangiopathy following renal ischaemia and revascularization: apoptosis of endothelial cells in action. PMID- 11477192 TI - Recovery from aplastic anaemia after institution of haemodialysis. PMID- 11477193 TI - Repair of femoral postcatheterization pseudoaneurysm and arteriovenous fistula with percutaneous implantation of endovascular stent. PMID- 11477194 TI - Pseudotuberculous pyelonephritis in a patient with autosomal dominant polycystic kidney disease undergoing long-term haemodialysis. PMID- 11477195 TI - Interferon and ribavirin therapy in dialysis patients with chronic hepatitis C. PMID- 11477198 TI - Genetic disorders of neonatal respiratory function. AB - Genetic risk for respiratory distress in infancy has been recognized with increasing frequency in neonatal intensive care units. Reports of family clusters of affected infants and of ethnic- and gender-based respiratory phenotypes point to the contribution of inheritance. Similarly, different outcomes among gestationally matched infants with comparable exposures to oxygen, mechanical ventilation, or nutritional deficiency also suggest a genetic risk for respiratory distress. Examples of inherited deficiency of surfactant protein B in both humans and genetically engineered murine lineages illustrate the importance of identifying markers of genetic risk. In contrast to developmental, inflammatory, or nutritional causes of respiratory distress that may resolve as infants mature, genetic causes result in both acute and chronic (and potentially irreversible) respiratory failure. The availability of clinically useful genetic markers of risk for respiratory distress in infancy will permit development of rational strategies for treatment of genetic lung disorders of infancy and more accurate counseling of families whose infants are at genetic risk for development of respiratory distress at birth or during early childhood. We review examples of genetic variations known to be associated with or cause respiratory distress in infancy. PMID- 11477199 TI - Pathogenesis of Shiga toxin-associated hemolytic uremic syndrome. AB - The aim of this review is to examine recent advances in experimental and clinical research relevant to the pathogenesis of diarrhea-associated hemolytic uremic syndrome with special reference to histopathologic findings, virulence factors of Shiga toxin-producing Escherichia coli, the host response, and the prothrombotic state. Despite significant advances during the past decade, the exact mechanism by which Shiga toxin-producing E. coli leads to hemolytic uremic syndrome remains unclear. Factors such as Shiga toxin, lipopolysaccharide, the adhesins intimin and E. coli-secreted proteins A, B, and D, the 60-MD plasmid, and enterohemolysin likely contribute to the pathogenesis. Data on the inflammatory response of the host, including leukocytes and inflammatory mediators, are updated. The pathogenesis of the prothrombotic state leading to thrombocytopenia secondary to endothelial cell damage and platelet activation is also discussed. A hypothetical sequence of events from ingestion of the bacteria to the development of full blown hemolytic uremic syndrome is proposed. PMID- 11477200 TI - Gadolinium chloride inhibits pulmonary macrophage influx and prevents O(2) induced pulmonary hypertension in the neonatal rat. AB - Newborn rats exposed to 60% O(2) for 14 d demonstrated a bronchopulmonary dysplasia-like lung morphology and pulmonary hypertension. A 21-aminosteroid antioxidant, U74389G, attenuated both pulmonary hypertension and macrophage accumulation in the O(2)-exposed lungs. To determine whether macrophage accumulation played an essential role in the development of pulmonary hypertension in this model, pups were treated with gadolinium chloride (GdCl(3)) to reduce lung macrophage content. Treatment of 60% O(2)-exposed animals with GdCl(3) prevented right ventricular hypertrophy (p < 0.05) and smooth muscle hyperplasia around pulmonary vessels, but had no effect on morphologic changes in the lung parenchyma. In addition, GdCl(3) inhibited 60% O(2)-mediated increases in endothelin-1, 8-isoprostane, and nitrotyrosine residues. Organotypic cultures of fetal rat distal lung cells were subjected to cyclical mechanical strain to assess the potential role of GdCl(3)-induced blockade of stretch-mediated cation channels in these effects. Mechanical strain caused a moderate increase of endothelin-1 (p < 0.05), which was unaffected by GdCl(3), but had no effect on 8 isoprostane or nitric oxide synthesis. A critical role for endothelin-1 in O(2) mediated pulmonary hypertension was confirmed using the combined endothelin receptor antagonist SB217242. We concluded that pulmonary macrophage accumulation, in response to 60% O(2), mediated pulmonary hypertension through up regulation of endothelin-1. PMID- 11477201 TI - Changes of CD14 and CD1a expression in response to IL-4 and granulocyte macrophage colony-stimulating factor are different in cord blood and adult blood monocytes. AB - Neonates are relatively immature in their immune response; thus, to further clarify the differences of monocyte function and differentiation between neonates and adults, we investigated their CD14(+)CD4(+) and CD14(+)CD16(+) monocyte subpopulations, production of IL-1beta and tumor necrosis factor-alpha induced by lipopolysaccharide, and their CD14 and CD1a phenotypic changes in response to IL 4 and granulocyte-macrophage colony-stimulating factor. Our results showed that 1) the expression of CD14 in cord blood monocytes was significantly lower than that in adult peripheral blood monocytes; 2) both the percentages of CD14(+)CD4(+) cells and CD14(+)CD16(+) cells among CD14(+) monocytes were also significantly lower in cord blood; 3) after stimulation by lipopolysaccharide for 72 h, production of both IL-1beta and tumor necrosis factor-alpha was lower in cord blood than that in adult peripheral blood; and 4) in response to IL-4 or GM CSF, the phenotype development of CD14 and CD1a in cord blood and adult peripheral blood was different. Down-regulation of CD14 expression in response to IL-4 and GM-CSF was slower in cord blood monocytes than that in adult peripheral blood monocytes. After 9 d of culture in the presence of IL-4 and GM-CSF, the percentage of CD1a(+) monocytes was significantly more increased in cord blood than that in adult peripheral blood. The reduced expression of CD14 and other mature phenotype markers such as CD16 and CD4 as well as the reduced IL-1beta and tumor necrosis factor-alpha production may contribute to the impaired immune response of neonates. Slower down-regulation of CD14 by IL-4 and GM-CSF suggests that differential properties of cord blood monocytes in response to cellular stress signals take a longer time than those of adult peripheral blood monocytes. PMID- 11477202 TI - Impaired glucocorticoid synthesis in premature infants developing chronic lung disease. AB - Premature infants have higher cortisol precursor concentrations than term infants; however, many sick preterm infants have surprisingly low cortisol concentrations. Those who develop chronic lung disease (CLD) have lower cortisol values than those who recover. We hypothesized that some infants have a decreased ability to synthesize cortisol, leading to physiologic disruptions including amplified inflammatory responses, thereby resulting in CLD. We measured cortisol, 11-deoxycortisol, 17-hydroxyprogesterone, 17-hydroxypregnenolone, dehydroepiandrosterone sulfate, and ACTH in 40 extremely low birth weight infants enrolled in a study of low-dose hydrocortisone therapy to prevent CLD. Thirty four infants survived and 15 developed CLD. Hydrocortisone therapy did not suppress ACTH or any measured steroid value. Before study (<48 h of life), 17-OH progesterone was higher in CLD infants, as was the ratio of 17-OH progesterone to 11-deoxycortisol. On d 15-19 (> or =72 h after end of therapy), basal and stimulated cortisol concentrations were lower in CLD infants. In contrast, the basal ratio of 11-deoxycortisol to cortisol was higher in CLD infants, as were stimulated values of 17-OH progesterone and stimulated ratios of 17-OH progesterone to 11-deoxycortisol and 11-deoxycortisol to cortisol. Thus, infants who developed CLD had lower basal and stimulated cortisol values, but elevated cortisol precursors and precursor to product ratios, compared with infants who recovered. These data support the hypothesis that these immature infants have a decreased capacity to synthesize cortisol, which may lead to a relative adrenal insufficiency in the face of significant illness. PMID- 11477203 TI - Developmental regulation of intestinal epithelial hydrolase activity in human fetal jejunal xenografts maintained in severe-combined immunodeficient mice. AB - Intestinal epithelial brush border hydrolases are important and sensitive enzyme markers of gastrointestinal development and function. Little is know about the mechanisms that regulate the induction of these enzymes during human fetal development, as these events occur primarily in utero. The present work used ectopically grafted human fetal jejunal xenografts (median age,13.3 wk of gestation), maintained in severe-combined immunodeficient mice, to study the differential expression of five different hydrolases after 10 wk of xenotransplantation. The spatio-temporal distribution of brush border alkaline phosphatase, aminopeptidase-N, alpha-glucosidase, lactase-phlorizin hydrolase, and dipeptidyl peptidase IV enzyme activities were measured quantitatively using scanning microdensitometry along the crypt-villus axes of fetal, xenograft, and pediatric (median age, 34 mo) biopsies. Ectopic grafting of fetal jejunum closely recapitulated the development of these enzymes in utero, with alkaline phosphatase, aminopeptidase-N, alpha-glucosidase, and dipeptidyl peptidase IV enzyme activities closely matching the spatio-temporal distribution and levels recorded in pediatric duodenal biopsies. Lactase-phlorizin hydrolase was the only enzyme not to reach values recorded in pediatric brush border membranes, although activities were significantly (5.6-fold) higher than in pretransplanted fetal bowel. Human jejunal xenografts therefore demonstrate an appropriate developmental induction of brush border hydrolase activity and may represent a useful model to study trans-acting factors that promote human epithelial differentiation and function in vivo. Characterization of such agents may be of potential therapeutic use in the treatment of diseases associated with gastrointestinal immaturity, notably necrotizing enterocolitis. PMID- 11477204 TI - Thromboxane A(2) limits differentiation and enhances apoptosis of cultured human trophoblasts. AB - Prostanoids influence differentiation in diverse cell types. Altered expression of cyclooxygenase and prostaglandins has been implicated in the pathophysiology of placental dysfunction, which results in preeclampsia and fetal growth restriction. We hypothesized that prostanoids modulate differentiation and apoptosis in cultured human trophoblasts. Villous cytotrophoblasts were isolated from term human placentas and cultured in serum-free medium. The level of human chorionic gonadotropin was used as a marker of biochemical differentiation of primary trophoblasts, and syncytia formation was used as a marker of morphologic differentiation. Of the prostanoids tested, we found exposure to thromboxane A(2) hindered both biochemical and morphologic differentiation of cultured trophoblasts. As expected, human chorionic gonadotropin levels in the media were elevated in a concentration-dependent manner in the presence of the thromboxane synthase inhibitor, sodium furegrelate, or the thromboxane A(2) receptor blocker SQ 29,548. Furthermore, thromboxane A(2) enhanced trophoblast apoptosis, determined using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining, cell morphology, and a concentration-dependent increase in p53 expression. We conclude that thromboxane A(2) hinders differentiation and enhances apoptosis in cultured trophoblasts from term human placenta. We speculate that thromboxane may contribute to placental dysfunction by restricting differentiation and enhancing apoptosis in human trophoblasts. PMID- 11477205 TI - Role of hypothalamic-pituitary axis in EGF action on maturation of adrenal gland in fetal rhesus monkey in vivo. AB - We determined the route of action of epidermal growth factor (EGF) [intraperitoneal (IP) versus intraamniotic administration] on adrenal development and whether its effects are mediated via the fetal hypothalamic-pituitary axis in the fetal rhesus monkey in vivo. EGF (40 microg) was administered IP (n = 9) or intraamniotic (n = 6) at 121, 123, 125, and 127 d gestation (term, approximately 165 +/- 10 d gestation). In addition, a competitive corticotropin-releasing factor antagonist ([D-phenylalanine(12), Norleucine(21,38)] corticotropin releasing factor(12-41) to block fetal pituitary ACTH secretion; 400 microg IP) and metyrapone (11beta-hydroxylase inhibitor to block adrenal cortisol synthesis; 15 mg IP and 15 mg intraamniotic) were administered, in combination with EGF (EGF+BLOCK; 40 microg IP; n = 4 fetuses). Control fetuses (n = 6) received saline injections in an equivalent volume. On gestational d 128, a hysterotomy was performed, and fetal adrenals were collected for morphometric analyses and immunocytochemical localization of 3beta -hydroxysteroid dehydrogenase (3betaHSD) and cytochrome P-450 11beta -hydroxylase/aldosynthase. Definitive zone (DZ) width and cortical width of 3betaHSD staining were significantly greater (p < 0.05) in the EGF IP-treated fetuses compared with controls and EGF+BLOCK. With EGF IP, 3betaHSD was increased in the DZ and induced extensively in the transitional zone of the fetal adrenal cortex, and cytochrome P-450 11beta-hydroxylase/aldosynthase immunoreactivity was induced to detectable levels in the DZ. The administration of EGF+BLOCK inhibited the expression of 3betaHSD in the transitional zone, but 3betaHSD expression was still increased in the DZ and cytochrome P-450 11beta hydroxylase/aldosynthase immunoreactivity was induced in the DZ. EGF intraamniotic administration had no significant effect on the width of the DZ or cortical width of 3betaHSD staining compared with controls. These data suggest that EGF acts via the hypothalamic-pituitary axis to modulate adrenal cortical growth and functional maturation of the transitional zone (the putative zona fasciculata), whereas EGF can act independently of the hypothalamic-pituitary axis to stimulate functional maturation of the DZ (the putative zona glomerulosa). PMID- 11477206 TI - Adenosine produces changes in cerebral hemodynamics and metabolism as assessed by near-infrared spectroscopy in late-gestation fetal sheep in utero. AB - Rises in fetal adenosine during hypoxia may have a metabolic inhibitory role that helps the fetus adapt to periods of low arterial partial pressure of oxygen (P(a)O(2)). We examined the fetal cerebral hemodynamic and metabolic responses to exogenous adenosine infusion and compared this with previous studies. Six fetal sheep at ca. 125 d gestation were instrumented under general anesthesia with catheters, flow probes, and near-infrared optodes and allowed to recover. After 3 d, adenosine was infused at a level known to reproduce fetal levels during hypoxia. Fetal hemodynamics and cerebral near-infrared spectroscopic (NIRS) variables were monitored and paired blood samples taken for oxygen delivery and consumption calculation. Fetal heart rate, mean arterial pressure, and carotid flow showed no change during adenosine infusion. Cerebral oxyhemoglobin (HbO(2)), deoxyhemoglobin (Hb), and blood volume rose, suggesting venous pooling in the brain. Cerebral cytochrome oxidase (CcO) became more oxidized, indicating reduction in electron flow down the mitochondrial electron transfer chain and, thus, a fall in metabolic rate. Blood sample analysis revealed that there was no change in oxygen delivery to the head but that cerebral oxygen consumption fell during adenosine infusion. These data indicate that fetal cerebral metabolism fell during infusion of adenosine at a level known to reproduce fetal plasma concentrations during hypoxia. PMID- 11477207 TI - Deleterious effects of IL-9-activated mast cells and neuroprotection by antihistamine drugs in the developing mouse brain. AB - Elevated mean IL-9 serum levels have been observed in human neonates who will later develop cerebral palsy. In earlier studies, using a newborn mouse model of excitotoxic lesions mimicking those described in human cerebral palsy, we found that IL-9 pretreatment exacerbated brain damage produced by intracerebral injections of the glutamatergic analog ibotenate. Among its different cell targets, the Th2 cytokine IL-9 is a mast cell growth and differentiation factor that can cause mast cells to release various substances including histamine. In the present study, we sought to determine whether the deleterious effects of IL-9 in our mouse model were mediated by mast cells through histamine release. All mouse pups were pretreated with intraperitoneal injections of IL-9 or saline between postnatal days (P) P1 and P5. Immunohistochemistry for murine mast cell protease-1 performed on P5 showed an increased density of labeled cells in the neopallium of IL-9-treated Swiss pups as compared with controls. Western blot analysis confirmed the increased murine mast cell protease-1 brain content of IL 9-treated Swiss mice. IL-9 pretreatment had no significant effect on ibotenate induced excitotoxic brain lesions in mast cell-deficient P5 pups (WBB6F1/J kit(W/W-v)), whereas IL-9 exacerbated these lesions in the control littermates with normal mast cell populations. Finally, cromoglycate or antihistamine drugs significantly reduced ibotenate-induced brain lesions in IL-9-treated Swiss pups. Taken together, these data suggest that recruitment of cerebral mast cells with histamine release may contribute to the exacerbation of neonatal excitotoxic brain lesions produced by IL-9. Neuroprotective strategies targeting mast cells may be useful in some neonates at risk for cerebral palsy. PMID- 11477208 TI - Neonatal asphyxia in rats: acute effects on cerebral kynurenine metabolism. AB - Two tryptophan metabolites, the anti-excitotoxic N-methyl-D-aspartate (NMDA) receptor antagonist kynurenic acid (KYNA) and the free radical generator 3 hydroxykynurenine (3-HK), have been proposed to influence neuronal viability in the mammalian brain. In rats, the brain content of both KYNA and 3-HK decreases immediately after birth, possibly to ensure normal postnatal functioning of NMDA receptors. Because complications of birth asphyxia have been suggested to be associated with anomalous NMDA receptor function, we examined the acute effects of an asphyctic insult on the brain levels of KYNA and 3-HK in neonatal rats. Asphyxia was induced in animals delivered by cesarean section on the last day of gestation, using the procedure introduced by Bjelke et al. (Brain Res 543: 1-9, 1991). KYNA and 3-HK levels were determined in the brain at seven time points between 10 min and 24 h after asphyxia. Up to 6 h, asphyxia caused 160-267% increases in KYNA levels. In the same tissues, 3-HK levels decreased (significantly at five of the seven time points), demonstrating an asphyxia induced shift in kynurenine pathway metabolism toward the neuroprotectant KYNA. This shift might constitute the brain's attempt to counter the ill effects of birth asphyxia. Furthermore, the transient increase in the brain KYNA/3-HK ratio in these animals might be causally related to the well-documented detrimental long-term effects of asphyxia. PMID- 11477209 TI - PDGF-beta receptor expression in the dorsocaudal brainstem parallels hypoxic ventilatory depression in the developing rat. AB - The temporal trajectory of platelet-derived growth factor (PDGF)-beta receptor activation within the dorsocaudal brainstem parallels that of the mild hypoxic ventilatory depression (HVD) seen in adult rats. We hypothesized that enhanced PDGF-beta receptor activity may account for the particularly prominent HVD of developing mammals. To study this issue, 2-, 5-, 10-, and 20-d-old rats underwent hypoxic challenges (10% O(2) for 30 min) after pretreatment with either vehicle (Veh) or the selective PDGF-beta receptor antagonist CGP57148B (intraperitoneal 100 mg/kg). The developmental characteristics and magnitude of the peak hypoxic ventilatory response (HVR) were not modified by the PDGF-beta receptor blocker. However, HVD was markedly attenuated by CGP57148B, and such effect, although still present, gradually abated with increasing postnatal age [p < 0.001, analysis of variance (ANOVA)]. Hypercapnic ventilatory responses were not affected by CGP57148B. The expression of PDGF-beta receptor in the dorsocaudal brainstem was assessed by immunoblotting and confirmed progressively decreasing expression with maturation. We conclude that PDGF-beta receptor activation during hypoxia is an important contributor to HVD at all postnatal ages but more particularly in the immature rat. PMID- 11477210 TI - Detection of cardiac hypertrophy in the fetus by approximation of the current dipole using magnetocardiography. AB - To determine the developmental changes in the myocardial current during fetal life, and to evaluate the clinical usefulness of magnetocardiography for prenatal diagnosis of cardiac hypertrophy or enlargement, we approximated the magnitude of the one-current dipole of the fetal heart using fetal magnetocardiography (fMCG). A total of 95 fetuses with gestational age of 20-40 wk were included in this study. fMCG was recorded with a nine-channel superconducting quantum interference device system in a magnetically shielded room. The magnitude of the dipole (Q) was calculated using an equation based on the fMCG amplitude obtained on the maternal abdomen and the distance between the maternal surface and fetal heart measured ultrasonographically. In uncomplicated pregnancies, the Q value correlated significantly with gestational age, reflecting an increase in the amount of myocardial current, i.e. myocardial mass. Moreover, the Q values in fetuses with cardiomegaly caused by various cardiovascular abnormalities tended to be higher than the normal values. Although there are some limitations of the methodology based on the half-space model, and fetal orientation may influence the magnitude of the dipole, making it smaller, fMCG recorded with a multichannel superconducting quantum interference device system is a clinically useful tool for noninvasive, prenatal, and electrical evaluation of fetal cardiac hypertrophy. PMID- 11477211 TI - Maturation-dependent differences in regulation of sarcoplasmic reticulum Ca(2+) ATPase in sheep myocardium in response to pressure overload: a possible mechanism for maturation-dependent systolic and diastolic dysfunction. AB - We have previously demonstrated that pressure-overload hypertrophy in adult sheep is associated with myocardial dysfunction whereas that in young lambs is associated with normal contractility. To probe for possible mechanisms of these age-dependent differences, we assessed mRNA expression of genes encoding critical components of myocardial Ca(2+) handling in the same animal model. We studied left ventricular myocardium of young and adult sheep with short-term (48 h) and long-term (6 wk) pressure overload induced by ascending aortic constriction. Six weeks of pressure overload induced the significant left ventricular hypertrophy (36 and 39% increase in left ventricular/body weight ratio in lambs and sheep, respectively). The Ca(2+) ATPase and Na(+)/Ca(2+) exchanger mRNA decreased with pressure overload only in the adult (p < 0.05). Ca(2+) channel mRNA was slightly increased by pressure overload regardless of age (p < 0.05). Calsequestrin, sarcoplasmic reticulum Ca(2+) release channel, or myosin heavy-chain mRNA levels did not significantly differ. In adult sheep after 6 wk of pressure overload, decreases in load-adjusted midwall shortening (systolic dysfunction) and prolongation of relaxation time constant (diastolic dysfunction) correlated with decreases in Ca(2+)-ATPase mRNA. The sarcoplasmic reticulum Ca(2+)-ATPase protein level and Ca(2+) uptake activity of isolated sarcoplasmic reticulum vesicles were depressed only in the adult with pressure-overload hypertrophy but not in the young. We demonstrated age-dependent differences in mRNA expression of Ca(2+) handling protein genes in response to pressure overload, which preceded the occurrence of hypertrophy and myocardial dysfunction. Thus, altered expression of Ca(2+)-handling protein genes may be one of the primary responses to pressure overload rather than a phenomenon secondary to myocardial hypertrophy. PMID- 11477212 TI - Glucose transporter type 1 deficiency syndrome (Glut1DS): methylxanthines potentiate GLUT1 haploinsufficiency in vitro. AB - Methylxanthines such as caffeine and theophylline are known to inhibit glucose transport. We have studied such inhibition in the glucose transporter type 1 deficiency syndrome (Glut1DS) by erythrocyte glucose transport assays. Data from four patients with individual mutations in the GLUT1 gene are discussed: patient 1 (hemizygosity), 3 (S66F), 15 (368Ins23), and 17 (R333W). Zero-trans influx of (14)C-labeled 3-O-methyl glucose (3-OMG) into erythrocytes of patients is reduced (patient 1, 51%; 3, 45%; 15, 31%; 17, 52%) compared with maternal controls. Inhibition studies on patients 1, 3, 17, and maternal controls show an IC(50) for caffeine of approximately 1.5 mM both in controls (n = 3) and patients (n = 3) at 5 mM 3-OMG concentration. In the same two groups, kinetic studies show that 3 mM caffeine significantly decreases V(max) (p < 0.005), whereas the decrease in K(m) is significant (p < 0.01) only in the three controls and one patient (patient 3). Kinetic data from individual patients permit us to speculate that the interactions between caffeine and Glut1 are influenced by the mutation. Three mM caffeine also inhibits the transport of dehydroascorbic acid (DHA), another substrate for Glut1. The combined effects of caffeine (3 mM) and phenobarbital (10 mM) on glucose transport, as determined in patient 15 and the maternal control, show no additive or synergistic inhibition. These data indicate that caffeine and phenobarbital have similar Glut1 inhibitory properties in these two subjects. Our study suggests that Glut1DS patients may have a reduced safety margin for methylxanthines. Consumption of methylxanthine-containing products may aggravate the neurologic symptoms associated with the Glut1DS. PMID- 11477213 TI - Enteral absorption of erythropoietin in the suckling rat. AB - Milk contains biologically relevant concentrations of erythropoietin (Epo), the primary hormone responsible for erythrocyte production. In animals, milk-borne Epo stimulates erythropoiesis. Epo receptors have been found in nonerythropoietic tissues including gastrointestinal tract. We hypothesized that milk-borne Epo is distributed to local gastrointestinal tissues, absorbed intact, and then distributed peripherally via the systemic circulation. Rat milk protected recombinant human Epo (rhEpo) from degradation in the suckling rat gastrointestinal tract. Simulated digestion of (125)I-rhEpo in suckling rat gastrointestinal juices was performed. When measured by acid precipitation and immunoassay, rat milk protected rhEpo from gastrointestinal juices better than saline (p < 0.0001). The fate of enterally administered milk-borne (125)I-rhEpo was examined in 10-d-old rats. RhEpo fed in rat milk was better protected from in vivo proteolytic degradation than rhEpo in saline (p < 0.05). After enteral (125)I-rhEpo dosing, radiolabeled protein from gastric tissue comigrated on SDS PAGE with intact rhEpo at 36.5 kD. To determine the local and systemic distribution of physiologic intakes of rhEpo, suckling rats were fed (125)I-rhEpo in rat milk, and tissues were harvested 1, 2, and 4 h later. Intact (125)I-rhEpo was found in gastric and small intestinal walls and lumens. Five percent of total administered dose was found intact in the plasma, whereas another 8 to 10% of total administered dose was localized to bone marrow, percentages comparable to those seen after parenteral administration. Radiolabel was also localized to liver and peripheral solid tissues. These patterns of localization and degradation of rhEpo after acute administration support both systemic absorption and gastrointestinal cellular processing. PMID- 11477214 TI - Interleukin-18 in human milk. AB - We analyzed IL-18 levels of human milk. Colostrum contained significantly higher levels of IL-18 compared with early milk and mature milk. By stepwise multiple linear regression analysis, preterm delivery and pregnancy complications of mothers significantly correlated with high levels of IL-18 in human milk (p = 0.0007 and 0.0018, respectively). There was a significant correlation between the levels of IL-18 and soluble Fas ligand in colostrum (p = 0.0003). IL-18 was detected in actively secreting epithelial cells in lactating mammary gland by immunohistochemical staining. These results suggest that IL-18 in colostrum plays an important role in host defense of high-risk neonates. PMID- 11477215 TI - Loop diuretics and in vitro relaxation of human fetal and newborn mouse airways. AB - This study was designed to test the hypotheses that furosemide directly causes relaxation in human fetal airway and that delivery of loop diuretics to either the adventitial or epithelial surface of newborn mouse airway results in equivalent relaxation. Isometric tension changes were measured in human fetal (11 16 wk) trachea and mainstem bronchus rings exposed to furosemide (300 microM) or saline after acetylcholine or leukotriene D(4) constriction. Significant decreases in isometric tension to furosemide were demonstrated after constriction with acetylcholine or leukotriene D(4). To examine the site of effect and mimic aerosolized and systemic administration, furosemide (3-300 microM) and bumetanide (0.3-30 microM) were applied separately to epithelial and adventitial surfaces of newborn mouse airways. No differences in airway diameter changes to epithelial or adventitial furosemide or bumetanide were observed, but a 10-fold difference in potency was found. In summary, human fetal airway relaxed to furosemide when constricted with either neurotransmitter or inflammatory mediator in vitro. Further, no differences in relaxation to equimolar epithelial and adventitial furosemide were observed in isolated newborn mouse airway. Taken together, this provides evidence that furosemide has a direct, nonepithelial-dependent effect on airway smooth muscle tone. PMID- 11477216 TI - A novel tool for the prediction of transmembrane protein topology based on a statistical analysis of the SwissProt database: the OrienTM algorithm. AB - OrienTM is a computer software that utilizes an initial definition of transmembrane segments to predict the topology of transmembrane proteins from their sequence. It uses position-specific statistical information for amino acid residues which belong to putative non-transmembrane segments derived from statistical analysis of non-transmembrane regions of membrane proteins stored in the SwissProt database. Its accuracy compares well with that of other popular existing methods. A web-based version of OrienTM is publicly available at the address http://biophysics.biol.uoa.gr/OrienTM. PMID- 11477217 TI - Metal ions modulate the plastic nature of Mycobacterium tuberculosis chaperonin 10. AB - Chaperonin-10s possess a highly flexible segment of approximately 10 residues that covers their dome-like structure and closes the central cavity of the chaperonin assembly. The dome loop is believed to contribute to the plasticity of their oligomeric structure. We have exploited the presence of a single tryptophan residue occurring in the dome loop of Mycobacterium tuberculosis chaperonin-10 (cpn-10), and through intrinsic fluorescence measurements show that in the absence of metal ions, the tryptophan is almost fully solvent exposed at neutral pH. The dome loop, however, assumes a closed conformation in the presence of metal ions, or at low pH. These changes are fully reversed in the presence of chelating agents such as EDTA, confirming the role of cations in modulating the metastable states of cpn-10. PMID- 11477218 TI - A critical view on conservative mutations. AB - By analysing the surface composition of a set of protein 3D structures, complemented with predicted surface compositional information for homologous proteins, we have found significant evidence for a layer composition of protein structures. In the innermost and outermost parts of proteins there is a net negative charge, while the middle has a net positive charge. In addition, our findings indicate that the concept of conservative mutation needs substantial revision, e.g. very different spatial preferences were found for glutamic acid and aspartic acid. The alanine screening often used in protein engineering projects involves the substitution of residues to alanine, based on the assumption that alanine is a "neutral" residue. However, alanine has a high negative correlation with all but the non-polar residues. We therefore propose the use of, for example, serine as a substitute for the residues that are negatively correlated with alanine. PMID- 11477219 TI - Loop fold nature of globular proteins. AB - Protein chains make numerous returns in globules, thus forming loops, closed by tight residue-to-residue contacts-closed loops. Previous statistical analysis of the sizes and locations of the closed loops in all major protein folds revealed that the loops have an almost standard contour length of 25-30 amino acid residues and follow one after another along the chain. In this work the closed loops of the major folds are presented in three dimensions. A special image filtering procedure is introduced that allows one to visualize the standard size closed loops for the first time. The loop positions along the sequences are verified by detection of loop-end clusters. PMID- 11477220 TI - Differences in conformational properties of the second intracellular loop (IL2) in 5HT(2C) receptors modified by RNA editing can account for G protein coupling efficiency. AB - Adenosine-to-inosine RNA editing events that have been demonstrated for 5HT (2C) receptors resulted in alterations of the amino acid sequence at positions 156, 158 and 160 in the intracellular loop 2 (IL2) region. The edited receptor isoforms were shown to have reduced basal activity, but similar maximum responses to agonist binding. To identify the molecular mechanism of these pharmacological effects of editing we explored the conformational properties of the edited IL2 in comparison with the wild type. The results from conformational studies of the IL2 isoforms, using biased Monte Carlo simulations with an implicit solvent model based on a screened Coulomb potential, show that the compared loops differ in their preferred spatial orientations as a result of differences in the conformational space that is accessible to them by energy criteria. For the IL2 of the unedited (5HT (2C-INI) ) receptor, the preference for structures oriented towards the 7TM bundle is larger than for the 5HT (2C-VGV) edited receptor. This difference in preferred orientation can affect the association of IL2 with other intracellular loop domains involved in G protein coupling and hence the coupling efficiency. The results illustrate the high sensitivity of the system to small changes in the interaction surface presented to other intracellular loops, and/or the G protein. PMID- 11477221 TI - How does heme axial ligand deletion affect the structure and the function of cytochrome b(562)? AB - We have recently generated a new mutant of cytochrome b(562) (cytb(562)) in which Met7, one of the axial heme ligands, is replaced by Ala (M7A cytb(562)). The M7A cytb(562) can bind heme and the UV-visible absorption spectrum is of a typical high-spin ferric heme. To investigate the effect of the lack of Met7 ligation on the structural integrity of cytb(562), thermal transition analyses of M7A cytb(562) were conducted. From the thermodynamic parameters obtained, it is concluded that the folding of M7A cytb(562) is comparable to the apoprotein despite the presence of heme. On the other hand, exogenous ligands such as cyanide and azide ions are readily bound to the heme iron, indicating that the axial coordination site is available for substrate binding. The peroxidase activity of this mutant is thus examined to evaluate new enzymatic function at this site and M7A cytb(562) was found to catalyze an oxidation reaction of aromatic substrates with hydrogen peroxide. These observations demonstrate that the Met7/His102 bis-ligation to the heme iron is crucial for the stable folding of cytb(562), whereas the functional conversion of cytb(562) is successfully achieved by the loose folding together with the open coordination site. PMID- 11477222 TI - Tolerance of point substitution of methionine for isoleucine in hen egg white lysozyme. AB - X-ray structure determination of proteins by using the multiple-wavelength anomalous dispersion method targeting selenomethionine is now widely employed. Isoleucine was examined for the second choice of the substitution of methionine next to leucine. We performed a systematic mutational study of the substitutions of methionine for isoleucine. All mutated lysozymes were less stable than the wild-type by about 1 kcal/mol and it is suggested that this instability was caused by the change in residual hydrophobicity from isoleucine to methionine. The X-ray structures of all mutant lysozymes were very similar to that of the wild-type. In addition, both the accessible surface areas and the conformation of the side chain of methionine in all mutant lysozymes were similar to those of the side chain at the respective isoleucine in the wild-type. Therefore, it is suggested that the mutation from isoleucine to methionine in a protein can be considered as a "safe" substitution. PMID- 11477223 TI - Antifibrinolytic effect of single apo(a) kringle domains: relationship to fibrinogen binding. AB - Elevated plasma concentrations of lipoprotein(a) [Lp(a)] are associated with an increased risk for the development of atherosclerotic disease which may be attributable to the ability of Lp(a) to attenuate fibrinolysis. A generally accepted mechanism for this effect involves direct competition of Lp(a) with plasminogen for fibrin(ogen) binding sites thus reducing the efficiency of plasminogen activation. Efforts to determine the domains of apolipoprotein(a) [apo(a)] which mediate fibrin(ogen) interactions have yielded conflicting results. Thus, the purpose of the present study was to determine the ability of single KIV domains of apo(a) to bind plasmin-treated fibrinogen surfaces as well to determine their effect on fibrinolysis using an in vitro clot lysis assay. A bacterial expression system was utilized to express and purify apo(a) KIV (2), KIV (7), KIV (9) DeltaCys (which lacks the seventh unpaired cysteine) and KIV (10) which contains a strong lysine binding site. We also expressed and examined three mutant derivatives of KIV (10) to determine the effect of changing critical residues in the lysine binding site of this kringle on both fibrin(ogen) binding and fibrin clot lysis. Our results demonstrate that the strong lysine binding site in apo(a) KIV (10) is capable of mediating interactions with plasmin modified fibrinogen in a lysine-dependent manner, and that this kringle can increase in vitro fibrin clot lysis time by approximately 43% at a concentration of 10 microM KIV (10). The ability of the KIV (10) mutant derivatives to bind plasmin-modified fibrinogen correlated with their lysine binding capacity. Mutation of Trp (70) to Arg abolished binding to both lysine-Sepharose and plasmin-modified fibrinogen, while the Trp (70) -->Phe and Arg (35) -->Lys substitutions each resulted in decreased binding to these substrates. None of the KIV (10) mutant derivatives appeared to affect fibrinolysis. Apo(a) KIV (7) contains a lysine- and proline-sensitive site capable of mediating binding to plasmin-modified fibrinogen, albeit with a lower apparent affinity than apo(a) KIV (10). However, apo(a) KIV (7) had no effect on fibrinolysis in vitro. Apo(a) KIV (2) and KIV (9) DeltaCys did not bind measurably to plasmin-modified fibrinogen surfaces and did not affect fibrinolysis in vitro. PMID- 11477224 TI - Prolonged display or rapid internalization of the IgG-binding protein ZZ anchored to the surface of cells using the diphtheria toxin T domain. AB - We have shown previously that the diphtheria toxin transmembrane domain (T) may function as a membrane anchor for soluble proteins fused at its C-terminus. Binding to membranes is triggered by acidic pH. Here, we further characterized this anchoring device. Soluble proteins may be fused at the N-terminus of the T domain or at both extremities, without modifying its membrane binding properties. This allows one to choose the orientation of the protein to be attached to the membrane. Maximum binding to the cell surface is reached within 1 h. Anchoring occurs on cells previously treated with proteinase K, suggesting that T interacts with the lipid phase of the membrane without the help of cell surface proteins. Binding does not permeabilize cells or affect cell viability, despite the fact that it permeabilizes liposomes and alters their structure. When attached to L929 fibroblasts, the proteins are not internalized and remain displayed at their surface for more than 24 h. When bound to K562 myeloid cells, the molecules are internalized and degraded. Thus, depending on the cell type, soluble proteins may be anchored to the surface of cells by the T domain for an extended time or directed towards an internalization pathway. PMID- 11477225 TI - Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris. AB - A custom-designed, highly hydrophilic gelatin was produced in Pichia pastoris. Secreted production levels in single-copy transformants were in the range 3-6 g/l of clarified broth and purification to near homogeneity could be accomplished by differential ammonium sulfate precipitation. Despite the fact that gelatins are highly susceptible to proteolysis because of their unfolded structure, the recombinant protein was shown to be fully intact by SDS-PAGE, N-terminal sequencing, gel filtration chromatography and mass spectrometry. Owing to its highly hydrophilic nature, the migration of the synthetic gelatin in SDS-PAGE was severely delayed. Esterification of the carboxylic amino acid side chains resulted in normal migration. The high polarity of the synthetic gelatin also accounts for its negligible surface activity in water at concentrations up to 5% (w/v), as determined by tensiometry. Circular dichroism spectrometry showed that the non-hydroxylated gelatin did not form triple helices at 4 degrees C. The spectrum was even more representative of the random coil conformation than the spectrum of natural non-hydroxylated gelatins. PMID- 11477226 TI - 2000 RSNA annual oration in diagnostic radiology: The future of interventional radiology. AB - Origins in imaging, procedural emphasis, and dependence on innovation characterize interventional radiology, which will continue as the field of image guided minimally invasive therapies. A steady supply of innovators will be needed. Current workforce shortages demand that this problem be addressed and in an ongoing fashion. Interventional radiology's major identity problem will require multiple corrective measures, including a name change. Diagnostic radiologists must fully embrace the concept of the dedicated interventionalist. Interspecialty turf battles will continue, especially with cardiologists and vascular surgeons. To advance the discipline, interventional radiologists must remain involved in cutting-edge therapies such as endograft repair of aortic aneurysms and carotid stent placement. As the population ages, interventionalists will experience a shift toward a greater emphasis on cancer treatment. Political agendas and public pressure will improve access to care and result in managed health care reforms. Academic centers will continue to witness a decline in time and resources available to pursue academic missions. The public outcry for accountability will result in systems changes aimed at reducing errors and process changes in the way physicians are trained, certified, and monitored. Evidence-based medicine will be the watchword of this century. Interventional radiology will maintain its role through development of methods for delivery of genes, gene products, and drugs to specific target sites; control of angiogenesis and other biologic processes; and noninvasive image-guided delivery of various forms of energy for ablation. PMID- 11477228 TI - Academic medicine: boom to bust. PMID- 11477229 TI - Radial artery mapping for coronary artery bypass graft placement. PMID- 11477230 TI - Computer network security for the radiology enterprise. AB - As computer networks become an integral part of the radiology practice, it is appropriate to raise concerns regarding their security. The purpose of this article is to present an overview of computer network security risks and preventive strategies as they pertain to the radiology enterprise. A number of technologies are available that provide strong deterrence against attacks on networks and networked computer systems in the radiology enterprise. While effective, these technologies must be supplemented with vigilant user and system management. PMID- 11477231 TI - Technologic advances in abdominal MR imaging. AB - Magnetic resonance (MR) imaging is finding an ever-growing role in the evaluation of a wide range of conditions in the abdomen. No longer confined to problem solving regarding abnormalities in solid organs, such as the liver and kidneys, MR imaging is increasingly being applied to the evaluation of the pancreatic and biliary ductal systems and even the bowel. Recent technical advances in hardware and software have allowed the acquisition of MR images that are largely free of artifact secondary to bowel peristalsis or respiratory motion; images providing excellent anatomic detail can now be obtained routinely. Faster sequences have reduced image acquisition time, thereby improving patient acceptance and allowing more efficient utilization of machine time. New three-dimensional sequences allow rapid image acquisition, reducing section misregistration and motion artifact while improving multiplanar reformations. The potential of MR imaging to provide functional and anatomic information is intriguing, and new techniques, including diffusion and perfusion imaging, are being evaluated. This review considers the advances in imaging hardware and pulse sequence design that underlie the increasing role of MR imaging in evaluation of the abdomen and discusses evolving clinical applications. PMID- 11477232 TI - Portal venous thrombosis or sclerosis in liver transplantation candidates: preoperative CT findings and correlation with surgical procedure. AB - PURPOSE: To review computed tomographic (CT) findings in patients with surgically proved portal venous (PV) thrombosis or sclerosis and to correlate these findings with the surgical procedure used at orthotopic liver transplantation (OLT). MATERIALS AND METHODS: Among 379 OLTs, PV thrombosis or sclerosis was found at surgery in 39 patients (10.3%). Before OLT, surgical records and CT images were retrospectively reviewed in 35 patients with available CT studies. Diameter of the extrahepatic PV and patency of the PV system were evaluated. Cavernous transformation, calcifications of the venous wall or thrombus, lesions suggestive of tumor, mesenteric varices, edema, or splenorenal shunt were recorded. A nonpaired Student t test and the Fisher exact test were used to analyze the results. RESULTS: Of 35 patients, 23 (66%) underwent thrombectomy with direct PV to-PV anastomosis and 12 (34%) had placement of venous grafts or other anastomoses. The extrahepatic PV was 8.2 mm, but it significantly (P < or = .05) decreased in patients with splenorenal shunt. In 30 patients, CT depicted thrombosis, PV calcification, or other abnormalities. The thrombus extended to or beyond the confluence of the splenic and superior mesenteric veins in 21 (60%) and 11 (31%) patients, respectively. Eleven patients (31%) had cavernous transformation of the PV; eight (23%), a cordlike sclerotic PV; 19 (54%), a splenorenal shunt; 11 (31%), PV calcification; 17 (49%), mesenteric edema; 14 (40%), mesenteric varices. Patients with a cordlike or calcified PV were significantly (P < or = .05) more likely to require modification of the surgical technique. CONCLUSION: Before OLT, CT can aid in assessment of PV and associated findings and in surgical management. PMID- 11477233 TI - Primary biliary cirrhosis: clinical, pathologic, and helical CT findings in 53 patients. AB - PURPOSE: To evaluate and compare clinical, pathologic, and helical computed tomographic (CT) findings of primary biliary cirrhosis (PBC). MATERIALS AND METHODS: The authors reviewed the medical records and CT scans of 53 patients who underwent evaluation, treatment, and orthotopic liver transplantation (OLT) at their institution. All patients underwent helical multiphase CT (total, 98 abdominal CT scans; range, one to five scans per patient). Multiple epidemiologic, clinical, and morphologic criteria were evaluated. Advanced disease was defined as hepatic insufficiency leading to OLT within the subsequent 2 years. Clinical and morphologic features were evaluated and compared in the advanced and less advanced cases of PBC. RESULTS: Common and characteristic findings included the following: 45 (85%) of the 53 patients were women with the onset of disease (diagnosis) in middle age (mean, 50.7 years; range, 26-71 years). The average time from diagnosis to OLT was 6.1 years (range, 1.5-20.0 years). CT findings in advanced PBC often resembled those seen in other forms of cirrhosis, with a small heterogeneously attenuating liver, varices, and splenomegaly. The liver in less advanced disease was usually enlarged or normal in size, with a smooth contour, little atrophy, and lacelike fibrosis and regenerative nodules in nearly one-third of the livers. Patients with less advanced disease frequently had varices (n = 33 [62%]) and ascites (n = 13 [24%]). Lymphadenopathy was seen in 47 (88%) patients. Hepatocellular carcinoma was found in four (8%) patients, two of whom also had chronic hepatitis C. During a follow-up period of 5-72 months (median, 46 months; mean, 42 months) after OLT, only two patients experienced recurrence of PBC. CONCLUSION: PBC is an important cause of liver failure, with distinctive clinical and CT findings that may assist diagnosis and allow adequate treatment. CT can demonstrate varices and ascites before frank cirrhosis is evident and can help evaluate the progression of the disease. PMID- 11477234 TI - Hemangioma-like lesions in chronic liver disease: diagnostic evaluation in patients. AB - PURPOSE: To quantify the risk of misdiagnosis of focal hepatic lesions manifesting at ultrasonography (US) as typical hemangiomas in a population at high risk for hepatocellular carcinoma (HCC) and to identify the most effective approach to their diagnostic evaluation. MATERIALS AND METHODS: A total of 1,982 patients with newly diagnosed cirrhosis underwent US and serum alpha-fetoprotein determinations for early detection of HCC. Focal lesions with typical features of hemangioma were evaluated with confirmatory findings of contrast material enhanced dynamic or spiral computed tomography (CT) and/or single photon emission CT with technetium 99m-labeled red blood cells and, in the absence of confirmatory imaging findings, US-guided fine-needle biopsy. Patients whose initial US scan depicted no lesions or hemangiomas were enrolled in a US follow up program. All hemangioma-like lesions detected during follow-up were evaluated, or biopsy was performed. RESULTS: US depicted hemangioma-like lesions in 44 of 1,982 patients: 22 hemangiomas and 22 HCCs. Hemangioma-like lesions detected during follow-up in 1,648 patients were HCCs (n = 22) or dysplastic nodules (n = 4). Only 85 (22%) of 383 patients with HCC had alpha-fetoprotein levels suggestive of the diagnosis. The probability of a diagnosis of HCC (or preneoplastic lesion) is 100% for hemangioma-like lesions depicted on subsequent US scans. CONCLUSION: If initial US examination of a cirrhotic liver depicts a hemangioma, confirmatory findings of imaging studies are necessary since 50% of hemangiomas in this study were hyperechogenic HCCs. US-guided biopsy can be safely performed, and its findings can be used to confirm the diagnosis. PMID- 11477235 TI - Strictures of the sigmoid colon: barium enema evaluation. AB - PURPOSE: To assess the accuracy of radiologic interpretation, in the absence of clinical information, in the differentiation of benign and malignant sigmoid strictures at barium enema examination. MATERIALS AND METHODS: On two occasions, four independent observers retrospectively assessed examination findings in 78 patients with documented sigmoid strictures (43 benign, 35 malignant). Each stricture was graded by using a five-point scale (definitely malignant to definitely benign). RESULTS: No significant difference existed between the areas under the receiver operating characteristic curves for the two assessments with any observer. Consensus findings indicated agreement among at least three of the four observers in 68 (87%) and 66 (85%) cases at the first and second assessments, respectively. One benign stricture was called malignant at both assessments. When consensus existed, the positive predictive value for malignant strictures was 96% at both assessments (sensitivity, 63% and 66%). Nine malignant strictures were called benign, three at both assessments. When consensus existed, the positive predictive value for benign strictures was 84% and 88% at the first and second assessments, respectively (sensitivity, 88% and 86%, respectively). CONCLUSION: The differentiation between a benign and a malignant sigmoid stricture can be made in most cases at barium enema examination. When a stricture appears malignant, the diagnosis is usually correct, but caution is advised when a stricture appears benign. PMID- 11477236 TI - Hepatocellular carcinoma: depiction of tumor parenchymal flow with intermittent harmonic power Doppler US during the early arterial phase in dual-display mode. AB - PURPOSE: To assess the effectiveness of contrast material-enhanced intermittent harmonic Doppler ultrasonography (US) in depicting tumor vessels and tumor parenchymal flow (stain) in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Fifty-eight patients with 65 HCC nodules were examined by using intermittent harmonic power Doppler US and digital subtraction harmonic B-mode US, both with intravenous administration of SH U 508A. Vascular findings at early arterial phase harmonic US were classified as positive enhancement or nonenhancement, depending on the tumor vascularity relative to the surrounding liver parenchyma. These results were compared with those of three-phase helical dynamic computed tomography (CT). RESULTS: For hypervascular HCCs, there was excellent depiction of tumor vessels and tumor stain with the two intermittent harmonic US methods. The sensitivity and specificity for depiction of tumor vascularity were 93% (41 of 44 nodules) and 100% (21 of 21), respectively, with intermittent harmonic power Doppler US and 86% (38 of 44) and 100% (21 of 21), respectively, with subtraction US, as compared with these values at dynamic CT. Attenuation was an important factor in the depictability of tumor vascularity at harmonic US. CONCLUSION: Contrast-enhanced intermittent harmonic US enables noninvasive demonstration of tumor vessels and especially tumor stain in HCC. PMID- 11477237 TI - Altered hepatic hemodynamics caused by temporary occlusion of the right hepatic vein: evaluation with Doppler US in 14 patients. AB - PURPOSE: To evaluate with Doppler ultrasonography (US) the altered hepatic hemodynamics caused by temporary occlusion of the right hepatic vein. MATERIALS AND METHODS: The study group consisted of 14 patients being considered for hepatic arterial infusion or transarterial embolization. In all patients, maximum peak velocity of the blood flow in the right portal vein was measured with Doppler US before and during the occlusion of the right hepatic vein. In 13 patients, color Doppler US was performed to evaluate Doppler signal in the portal venous branch in the occluded area before and during occlusion. Average peak velocity in the right hepatic artery in eight patients was measured by using a transducer-tipped guide wire before and during occlusion. RESULTS: Maximum peak velocity of the right portal vein significantly decreased with occlusion (P <.01). Hepatic venous occlusion changed the Doppler signal in the portal venous branch in the occluded area from hepatopetal to no signal in 10 patients; to weakened hepatopetal in two; and to hepatofugal in one. Average peak velocity of the right hepatic artery showed a decrease or plateau for 15-30 seconds after the start of occlusion and then a rapid increase to reach a plateau at around 75-90 seconds, with 1.5-2 times as much velocity as that before occlusion. CONCLUSION: Increase in hepatic arterial velocity is accompanied by a decrease in the portal velocity with temporary occlusion of the right hepatic vein; the expected increased drainage through the portal vein was almost undetectable. PMID- 11477238 TI - Penetrating stab wounds to the abdomen: use of serial US and contrast-enhanced CT in stable patients. AB - PURPOSE: To evaluate the usefulness of computed tomography (CT) and ultrasonography (US) for the initial assessment of penetrating abdominal stab wounds in patients who presented to the emergency department without indication for immediate laparotomy. MATERIALS AND METHODS: During 36 months, 32 patients with a penetrating stab wound to the abdomen were examined with serial US (at admission and 12 hours later) and helical CT, with contrast material administered orally, intravenously, and rectally. Presence of hemoperitoneum and integrity of solid and hollow viscera were evaluated with both methods. Sonograms were interpreted by the radiologist who performed the examination, and CT images were independently evaluated by two radiologists. Findings of both techniques were compared with clinical outcome and/or surgical findings. RESULTS: One (3.1%) of 32 patients required surgery: Surgical findings were massive hemoperitoneum and an extensive hepatic laceration. Both US and CT depicted these abnormalities. Thirty-one (96.9%) patients were treated conservatively, without surgery, and remained asymptomatic during 28 days of clinical follow-up after discharge from the hospital. US and/or CT showed intraperitoneal abnormalities in 21 of these patients. In 11 patients, both methods showed no evidence of visceral injury or hemoperitoneum, and none of these patients required surgery. CONCLUSION: Serial US and CT help guide treatment for stable patients with penetrating stab injuries to the abdomen. PMID- 11477239 TI - Whole-body 18F dopa PET for detection of gastrointestinal carcinoid tumors. AB - PURPOSE: To evaluate fluorine 18 (18F) dopa positron emission tomography (PET) in comparison with established imaging procedures in gastrointestinal carcinoid tumors. MATERIALS AND METHODS: After evaluation of the normal distribution of 18F dopa, 17 patients with histologically confirmed tumors were examined with 18F dopa PET. Results of 2-[fluorine 18]fluoro-2-deoxy-D-glucose (FDG) PET, somatostatin-receptor scintigraphy, and morphologic imaging (computed tomography and/or magnetic resonance imaging) were available for all patients. Results of the procedures were evaluated by two radiologists and two nuclear medicine specialists, whose consensus based on all available histologic, imaging, and follow-up findings was used as the reference standard. RESULTS: Ninety-two tumors were diagnosed: eight primary tumors, 47 lymph node metastases, and 37 organ metastases. 18F dopa PET led to 60 true-positive findings (seven primary tumors, 41 lymph node metastases, 12 organ metastases); FDG PET, 27 (two primary tumors, 14 lymph node metastases, 11 organ metastases); somatostatin-receptor scintigraphy, 52 (four primary tumors, 27 lymph node metastases, 21 organ metastases); and morphologic imaging, 67 (two primary tumors, 29 lymph node metastases, 36 organ metastases). This resulted in the following overall sensitivities: 18F dopa PET, 65% (60 of 92); FDG PET, 29% (27 of 92); somatostatin-receptor scintigraphy, 57% (52 of 92); morphologic procedures, 73% (67 of 92). Although the morphologic procedures were most sensitive for organ metastases, 18F dopa PET enabled best localization of primary tumors and lymph node staging. CONCLUSION: 18F dopa PET is a promising procedure and useful supplement to morphologic methods in diagnostic imaging of gastrointestinal carcinoid tumors. PMID- 11477240 TI - Use of 99mTc (V) DMSA scintigraphy in the detection and localization of intestinal inflammation: comparison of findings and colonoscopy and biopsy. AB - PURPOSE: To evaluate the potential use of technetium 99m (99mTc) (V) dimercaptosuccinic acid (DMSA) scintigraphy in the detection and localization of intestinal inflammation. MATERIALS AND METHODS: In a prospective study, 62 patients who were suspected of having intestinal inflammation and 30 control subjects were enrolled. All patients underwent 99mTc (V) DMSA scintigraphy and colonoscopy with biopsy within 1 week. 99mTc (V) DMSA scintigrams were interpreted blindly with respect to clinical information, and radiotracer uptake in the bowel segments was graded. The findings were then compared with the results of the colonoscopy and colonoscopic biopsy. RESULTS: In the detection of intestinal inflammation, findings at 99mTc (V) DMSA scintigraphy were as follows: true-positive in 55, false-positive in two, true-negative in 32, and false negative in three. Overall sensitivity was 95%; overall specificity, 94%; and overall accuracy, 95%. CONCLUSION: Our results show that 99mTc (V) DMSA scintigraphy is a useful noninvasive diagnostic test for the detection and localization of intestinal inflammation. PMID- 11477241 TI - CT colonography with teleradiology: effect of lossy wavelet compression on polyp detection--initial observations. AB - PURPOSE: To assess the consequences of lossy compression on the diagnostic accuracy of CT colonography for detecting colonic polyps. MATERIALS AND METHODS: Helical CT images of cleansed colonic segments were evaluated. Source images were compressed to 1:1, 10:1, and 20:1 ratios with lossy wavelet compression. Two independent readers blinded to corresponding colonoscopic results analyzed 144 randomly ordered colonic segments in multiplanar and volume-rendered endoscopic views. Sensitivity, specificity, and receiver operating characteristic curves were generated for each compression ratio on the basis of expressed confidence in lesion presence. Similar analyses were performed to assess distention and bowel preparation adequacy and evaluation time required. RESULTS: Results based on video colonoscopy-confirmed lesions revealed 100% (four of four) sensitivity for lesions larger than 10 mm for compression ratios 1:1, 10:1, and 20:1 for both readers; sensitivities for all lesions smaller than 10 mm were 50%-78%, 38%-67%, and 38%-67% for respective ratios for both readers. Differences in diagnostic performance for each reader across ratios were not significant (P =.30-.99, McNemar test). The time required to evaluate and assess bowel preparation and distention adequacy did not change significantly across ratios. CONCLUSION: On the basis of the patient sample, lossy compression of transverse source images to at least a 20:1 ratio did not adversely affect diagnostic performance or evaluation time for CT colonography. PMID- 11477242 TI - Influence of imaging on clinical decision making in the treatment of lower back pain. AB - PURPOSE: To assess the impact of cross-sectional imaging with magnetic resonance (MR) imaging or computed tomography (CT) on clinical decision making for patients with lower back pain (LBP). MATERIALS AND METHODS: A randomized controlled before and-after study was performed in 145 patients who had symptomatic lumbar spinal disorders and had been referred to orthopedists or neurosurgeons. Participants were a subgroup within a multicenter pragmatic randomized comparison of two imaging policies on LBP treatment: "imaging" versus "no imaging," unless a clear indication developed. Paired assessments were made of diagnosis, diagnostic confidence, proposed treatment, treatment confidence at trial entry and follow up, and expectations of imaging. Data were analyzed according to the groups as randomized. RESULTS: At follow-up, there were no statistically significant differences between the groups with respect to diagnosis or treatment plans. Significant increases in diagnostic and therapeutic confidence between trial entry and follow-up were observed for both groups, with a significantly greater increase in diagnostic confidence (P =.01) in the imaging group. CONCLUSION: Imaging may increase diagnostic confidence but has minimal influence on diagnostic or therapeutic decisions for patients with LBP. The results highlight the need for evidence-based guidelines for imaging in LBP treatment. PMID- 11477243 TI - Quantitative US of the calcaneus: cutoff levels for the distinction of healthy and osteoporotic individuals. AB - PURPOSE: To calculate cutoff levels for quantitative ultrasonography (US) performed to distinguish healthy individuals and those with osteoporosis identified with dual x-ray absorptiometry. MATERIALS AND METHODS: In 1,357 patients (856 females, aged 55.1 years +/- 15.4 [mean +/- SD]; 501 males, aged 50.4 years +/- 15), bone mineral density measurements of the lumbar spine (posteroanterior, L1 through L4) and femoral neck were obtained, and quantitative US was performed to determine the stiffness of the calcaneus. Individuals with a T score less than -2.5 (osteoporotic) at the spine and femur were identified, and upper T-score cutoff values (3 SDs from the mean) in the groups of male and female patients with osteoporosis were identified. RESULTS: Females with dual x ray absorptiometric values that were indicative of osteoporosis of the spine had an upper T-score cutoff value of -1.0 (males, -0.2). Females who had femoral osteoporosis showed an upper quantitative US T-score cutoff value of -0.6 (males, 0). CONCLUSION: Cutoff values may permit the use of quantitative US to screen for the presence of osteoporosis in the spine and femur. Quantitative US will help to prevent unnecessary dual x-ray absorptiometric and conventional radiographic examinations. PMID- 11477244 TI - Full- versus partial-thickness Achilles tendon tears: sonographic accuracy and characterization in 26 cases with surgical correlation. AB - PURPOSE: To determine the accuracy of the use of sonography for differentiation of full- from partial-thickness tears or tendinosis of the Achilles tendon by using surgical findings as the standard of reference and to identify sonographic characteristics of full-thickness tears that can be used to differentiate the two types of tears. MATERIALS AND METHODS: In part A of this study, sonographic findings (based on reports) in 26 consecutive cases of tears of the Achilles tendon were compared with surgical findings. In part B, the sonograms were blindly and retrospectively evaluated with respect to six sonographic characteristics possibly related to pathologic findings in the tendon, and the characteristics were correlated with surgical findings. RESULTS: In part A, statistical data regarding the use of sonographic findings to distinguish full- from partial-thickness tears were as follows: sensitivity, 100%; specificity, 83%; accuracy, 92%; positive predictive value, 88%; and negative predictive value, 100%. In part B, tendon thickness (P <.001), posterior acoustic shadowing (P =.007), and tendon retraction (P <.001) were correlated with full-thickness tears. Visualization of fat herniation (P =.051) and of the plantaris tendon (P =.098) demonstrated marginal correlation with full-thickness tears. Echogenicity at the site of the pathologic finding in the tendon showed no significant correlation. CONCLUSION: Sonography can be used to differentiate full- from partial-thickness tears or tendinosis of the Achilles tendon with 92% accuracy. Undetectable tendon at the site of injury, tendon retraction, and posterior acoustic shadowing demonstrate statistically significant correlation with full thickness tears. PMID- 11477245 TI - Delayed muscle injuries in arterial insufficiency: contrast-enhanced MR imaging and 31P spectroscopy in rats. AB - PURPOSE: To evaluate whether the vascular system resulting from an arterial lesion shows differences in permeability to a tracer with respect to the normal vascular system and whether eventual differences are maintained for long periods. MATERIALS AND METHODS: Permanent ischemia was induced in rats with femoral arterial removal, and magnetic resonance (MR) imaging was performed after 1, 7, 14, and 90 days. Gadopentetate dimeglumine was injected, and the kinetics of its penetration in the leg were studied. Phosphorus 31 spectroscopy was performed to determine the bioenergetic characteristics of the gastrocnemius muscle at rest and stimulation. Ischemic muscles were then processed for electron microscopy. RESULTS: After ischemia induction, a hyperintense area that progressively decreased was present on T2-weighted images. Gadopentetate dimeglumine improved the signal intensity of the area. Three months after arterial occlusion, the contrast-enhanced images still showed microvessels highly permeable to the tracers. Spectroscopic data revealed that 3 months after arterial removal, the bioenergetic reserve of the gastrocnemius muscle was reduced, suggesting that the contrast-enhanced MR imaging-visible area is functionally relevant. Ultrastructural examination revealed persistent muscle damage and signs of chronic microangiopathy. CONCLUSION: After ischemia induction, the restitutio ad integrum is not complete, and delayed muscle injuries can result from arterial insufficiency. PMID- 11477246 TI - Percutaneous tumor ablation: increased necrosis with combined radio-frequency ablation and intratumoral doxorubicin injection in a rat breast tumor model. AB - PURPOSE: To determine whether a combination of intratumoral doxorubicin injection and radio-frequency (RF) ablation increases tumor destruction compared with RF ablation alone in an animal tumor model. MATERIALS AND METHODS: R3230 mammary adenocarcinoma 1.2-1.5-cm- diameter nodules (n = 110) were implanted subcutaneously in 84 female Fischer rats. For initial experiments (n = 46), tumors were treated with (a) conventional, monopolar RF (250 mA +/- 25 [SD] at 70 degrees C +/- 1 for 5 minutes) ablation alone; (b) direct intratumoral doxorubicin injection (volume, 250 microL; total dose, 0.5 mg) alone; (c) combined therapy (doxorubicin injection immediately followed by RF ablation); (d) RF ablation and injection of 250 microL of distilled water; or (e) no treatment. In subsequent experiments, amount of doxorubicin (0.02-2.50 mg; n = 40 additional tumors) and timing of doxorubicin administration (2 days before to 2 days after RF ablation; n = 24 more tumors) were varied. Pathologic examination, including staining for mitochondrial enzyme activity and perfusion, was performed, and the resultant tumor destruction from each treatment was evaluated. RESULTS: Coagulation diameter was 6.7 mm +/- 0.6 for tumors treated with RF ablation alone and 6.9 mm +/- 0.7 for those treated with RF ablation and water (P =.52), while intratumoral doxorubicin injection alone produced only 2.0-3.0 mm of coagulation (P <.001). Increased coagulation was observed only with combined doxorubicin injection and RF therapy (P <.001). Coagulation was dependent on concentration and timing of doxorubicin administration, with greatest coagulation (11.5 mm +/- 1.1) observed for doxorubicin administered within 30 minutes of RF ablation. CONCLUSION: Adjuvant intratumoral doxorubicin injection increases coagulation in solid tumors compared with RF ablation alone. Increased tumor destruction is also seen when doxorubicin is administered after RF ablation, which suggests that RF ablation may sensitize tumors to chemotherapy. Such combination therapies may, therefore, offer improved methods for ablating solid tumors. PMID- 11477247 TI - In vitro effects of transcatheter injection on structure, cell viability, and cell metabolism in fibroblast-impregnated alginate microspheres. AB - PURPOSE: To determine if microsphere-encapsulated cell preparations can be delivered through a microcatheter without compromising microsphere structure, cell viability, or metabolism. MATERIALS AND METHODS: Fibroblast-impregnated microspheres were fabricated by using 1.0% alginate and rabbit synovial fibroblasts. Fibroblast-impregnated alginate microspheres injected through microcatheters were analyzed in parallel with identical noninjected microspheres. The effects of transcatheter injection on structure and cell viability (percentage of viable cells per microsphere) were correlated with microsphere size. Structural effects were analyzed by using light microscopy, and 7-day percentage (ratio of live cells to dead cells) cell viability was assessed with confocal microscopy and fluorescent staining. In a second series of experiments, the metabolism of small microspheres was studied during a course of 7 days by using a spectrophotometric bioanalyzer. RESULTS: Transcatheter injection caused fracturing and/or fragmentation of large (800-1,000 microm) and medium (500-750 microm) microspheres, while small (250-400 microm) microspheres were structurally unaffected by transcatheter injection. Fracturing and fragmentation were associated with cell release from the alginate matrix. Although transcatheter injection reduced cell viability by 17%-23% in all size categories, it did not cause a detectable alteration in the rate of glucose metabolism. CONCLUSION: Transcatheter injection was physiologically well tolerated by fibroblasts encapsulated in alginate microspheres; however, when microsphere diameter exceeded the catheter diameter, fracturing and fragmentation of microspheres compromised the sequestration function of the microsphere vector. PMID- 11477248 TI - Occlusive myocardial infarction: investigation of bis-gadolinium mesoporphyrins enhanced T1-weighted MR imaging in a cat model. AB - PURPOSE: To test whether bis-gadolinium mesoporphyrins-enhanced magnetic resonance (MR) imaging can accurately depict irreversibly damaged myocardium in occlusive myocardial infarction. MATERIALS AND METHODS: Ten cats were subjected to 90 minutes of occlusion of the left anterior descending coronary artery. Bis gadolinium mesoporphyrins-enhanced T1-weighted MR imaging was performed in the cats for 6 hours. Histopathologic examinations with 2'3'5-triphenyl tetrazolium chloride (TTC) staining and electron microscopy were performed on the resected specimens. The time course and pattern of signal intensity enhancement were evaluated. The size of the infarcted myocardium was estimated on the MR images by measuring the size of the signal intensity-enhanced area. RESULTS: In eight of 10 cats, it was impossible to distinguish infarcted myocardium from normal myocardium at visual inspection of T1-weighted MR images. The contrast ratio between infarcted and normal myocardium did not increase significantly over time. In one of the two remaining cats, a doughnut pattern of signal intensity enhancement was noted. The other cat showed intensely homogeneous enhancement of infarcted myocardium at MR imaging. The size of the area of signal intensity enhancement at MR imaging in these two cats was accurately mapped to that of the infarction on the TTC-stained specimens. CONCLUSION: Occlusive myocardial infarction cannot be accurately detected at bis-gadolinium mesoporphyrins enhanced MR imaging. PMID- 11477249 TI - Mr flow measurement in the internal mammary artery-to-coronary artery bypass graft: comparison with graft stenosis at radiographic angiography. AB - PURPOSE: To evaluate the sensitivity and specificity of breath-hold magnetic resonance (MR) flow measurement for detection of significant stenosis in internal mammary artery bypass grafts. MATERIALS AND METHODS: Twenty-six consecutive patients who had undergone coronary artery bypass surgery were examined. Breath hold velocity-encoded cine MR images were obtained at the midpoint of the internal mammary artery between its origin from the subclavian artery and the distal anastomosis to the left anterior descending artery. RESULTS: MR images were obtained successfully in 24 patients. At conventional angiography, no significant stenosis was observed in 17 patients (group A), and significant stenosis (diameter > 70%) was observed in seven patients (group B). The mean diastolic-to-systolic peak velocity ratio in group B (0.61 +/- 0.44 [SD]) was significantly lower than that in group A (1.88 +/- 0.96; P <.01). Evaluation of graft stenosis with the diastolic-to-systolic peak velocity ratio revealed a sensitivity of 86% and a specificity of 88%. The mean blood flow rate at baseline in group B (16.9 mL/min +/- 5.5) was significantly lower than that in group A (79.8 mL/min +/- 38.2; P <.01). The sensitivity and specificity of MR blood flow measurement in predicting significant stenosis were 86% and 94%, respectively. The mean pharmacologic flow reserve ratios were 2.00 +/- 1.43 in group A and 1.39 +/- 1.46 in group B (P >.05). CONCLUSION: Fast MR blood flow measurement at baseline is highly useful for predicting significant stenosis in internal mammary arterial grafts. PMID- 11477250 TI - Repeatability of quantitative CT indexes of emphysema in patients evaluated for lung volume reduction surgery. AB - PURPOSE: To evaluate the repeatability of quantitative computed tomographic (CT) indexes of emphysema and the effect of spirometric gating of lung volume during CT in candidates for lung volume reduction surgery (LVRS). MATERIALS AND METHODS: Initial and same-day repeat routine inspiratory spiral chest CT studies were performed in 29 LVRS candidates (group 1, routine study vs repeat study). In a separate cohort of 29 LVRS candidates, spiral chest CT studies were performed both without and with spirometric gating by using a spirometer to trigger scanning at 90% of vital capacity (group 2, spirometric gating study). In each study, Pearson and intraclass correlation coefficients were calculated to determine the agreement between multiple pairs of whole-lung quantitative CT indexes of emphysema, and mean values were compared with two-tailed paired t tests. RESULTS: Pearson and intraclass correlation coefficients were high for all quantitative CT indexes (all > or = 0.92). No significant differences were found between mean values of quantitative CT indexes in group 1. Variation in quantitative CT results was small but more prominent in group 2 than in group 1. The variation in quantitative CT results was primarily related to differences in lung volume (r(2) as great as 0.83). CONCLUSION: Repeatability of quantitative CT test results in LVRS candidates is high and unlikely to improve by using spirometric gating. PMID- 11477251 TI - Post-lung transplantation bronchiolitis obliterans syndrome: usefulness of expiratory thin-section CT for diagnosis. AB - PURPOSE: To assess the usefulness of thin-section expiratory computed tomography (CT), as compared with that of thin-section inspiratory CT, in detecting airway obstruction and air trapping in pediatric lung transplant recipients with bronchiolitis obliterans syndrome (BOS). MATERIALS AND METHODS: Thin-section CT scans were obtained at full inspiration and end expiration in 21 pediatric lung transplant recipients with proved BOS and in 41 transplant recipients with normal airways. True diagnosis was based on pulmonary function test results. Inspiration CT scans were scored for extent of decreased attenuation of the lung parenchyma; expiration CT scans were scored for extent of air trapping. RESULTS: The sensitivity of inspiratory CT for enabling diagnosis of BOS was 71%; the specificity, 78%; the positive predictive value, 62%; and the negative predictive value, 84%. The sensitivity of expiratory CT for enabling diagnosis of BOS was 100%; the specificity, 71%; the positive predictive value, 64%; and the negative predictive value, 100%. Expiratory CT scores correlated more strongly (rho = 0.75, P <.01) with pulmonary function test-based scores than did inspiratory CT scores (rho = 0.48, P <.01). Nominal logistic regression analysis revealed that expiratory CT was a more powerful predictor of true diagnosis (P <.01) than was inspiratory CT (P =.10). CONCLUSION: Expiratory CT is sensitive for depicting BOS related airway abnormalities and may be more useful than inspiratory CT for diagnosis of small airway obstruction. PMID- 11477252 TI - The double bubble sign. PMID- 11477253 TI - Cyclosporin A-induced fibroadenomas of the breast: report of five cases. AB - Five female patients undergoing cyclosporin A therapy had newly developed breast masses. Masses were bilateral in three of the five patients and palpable in four patients. The imaging findings were suggestive of fibroadenomas, and biopsy results were used to confirm the diagnosis. With the development of new breast lesions in patients after transplantation surgery, the diagnosis of cyclosporin A induced fibroadenomas should be considered. PMID- 11477255 TI - Case 37: Juxtacaval fat collection--mimic of lipoma in the subdiaphragmatic inferior vena cava. PMID- 11477256 TI - Prediction of aortoiliac stent-graft length: comparison of measurement methods. AB - PURPOSE: To determine the accuracy of helical computed tomography (CT), projectional angiography derived from CT angiography, and intravascular ultrasonographic withdrawal (IUW) length measurements for predicting appropriate aortoiliac stent-graft length. MATERIALS AND METHODS: Helical CT data from 33 patients were analyzed before and after endovascular repair of abdominal aortic aneurysm (Aneuryx graft, n = 31; Excluder graft, n = 2). The aortoiliac length of the median luminal centerline (MLC) and the shortest path (SP) that remained at least one common iliac arterial radius away from the vessel wall were calculated. Conventional angiographic measurements were simulated from CT data as the length of the three-dimensional MLC projected onto four standard viewing planes. These predeployment lengths and IUW length, available in 24 patients, were compared with the aortoiliac arterial length after stent-graft deployment. RESULTS: The mean error values of SP, MLC, the maximum projected MLC, and IUW were -2.1 mm +/- 4.6 (SD) (P =.013), 9.8 mm +/- 6.8 (P <.001), -5.2 mm +/- 7.8 (P <.001), and 14.1 mm +/- 9.3 (P <.001), respectively. The preprocedural prediction of the postprocedural aortoiliac length with the SP was significantly more accurate than that with the MLC (P <.001), maximum projected MLC (P <.001), and IUW (P <.001). CONCLUSION: The shortest aortoiliac path length maintaining at least one radius distance from the vessel wall most accurately enabled stent-graft length prediction for 31 AneuRx and two Excluder stent-grafts. PMID- 11477257 TI - Renal time-resolved MR angiography: quantitative comparison of gadobenate dimeglumine and gadopentetate dimeglumine with different doses. AB - PURPOSE: Results with different doses of gadobenate dimeglumine and gadopentetate dimeglumine were compared at magnetic resonance (MR) angiography of the renal arteries. The signal-to-noise ratio (SNR) was evaluated as a quantitative measure of image quality. MATERIALS AND METHODS: Sixty consecutive patients (age range, 24-81 years; mean age, 65 years) underwent intraarterial digital subtraction angiography (DSA) and contrast material-enhanced time-resolved MR angiography. DSA was the standard of reference. Fifteen patients received gadopentetate dimeglumine at doses of 0.2 or 0.1 mmol per kilogram of body weight. Fifteen patients received gadobenate dimeglumine at doses of 0.05 or 0.1 mmol/kg. The SNR was calculated in the aorta and both main renal arteries. The number and degree of stenoses of the renal arteries and accessory vessels were evaluated by four observers. RESULTS: SNRs with gadobenate dimeglumine at a dose of 0.1 mmol/kg were significantly superior to those with gadopentetate dimeglumine at a dose of 0.1 mmol/kg. Differences were not statistically significant between the SNRs in the other groups. Eleven (85%) of 13 hemodynamically significant renal artery stenoses were detected correctly with MR angiography as were 22 (85%) of 26 accessory renal arteries. CONCLUSION: SNRs with gadobenate dimeglumine were higher than those with gadopentetate dimeglumine, but in most cases the differences in SNRs were not statistically significant. PMID- 11477258 TI - Celiac-bimesenteric trunk: anatomic and radiologic description--case report. AB - The authors report the case of a 39-year-old man with a common origin of three arteries-the celiac, superior mesenteric, and inferior mesenteric arteries-that has not been described previously in the literature, to their knowledge. This variant, which they call the celiac-bimesenteric trunk, is documented with a selective angiogram, and an embryologic explanation is offered. PMID- 11477259 TI - Hospital costs for elective endovascular and surgical repairs of infrarenal abdominal aortic aneurysms. AB - PURPOSE: To determine and compare the average in-hospital costs of elective open surgical and endovascular repairs of infrarenal abdominal aortic aneurysms. MATERIALS AND METHODS: Total actual cost data for patients undergoing elective endovascular (n = 181) or open surgical (n = 273) repair of abdominal aortic aneurysms between 1997 and 1999 were retrieved. The mean total hospital cost (including stent-graft costs and excluding attending physician fees) and mean postoperative length of stay were calculated for each treatment group. Costs were expressed in 1999 U.S. dollars. RESULTS: Endovascular repair yielded a shorter postoperative length of stay than did open surgery (mean stay, 3.4 vs 8.0 days; P <.001) and a lower proportion of patients who were admitted to the intensive care unit for 1 full day or longer (2.8% vs 36.3%; P <.001). The mean total hospital cost was significantly higher for endovascular repair than for open surgery ($20,716 vs $18,484; P <.001). CONCLUSION: Hospital costs were higher for endovascular repair than for open surgical repair. However, endovascular repair was associated with a decreased length of stay and fewer intensive care unit admissions. The increased mean hospital cost for endovascular repair was smaller than one would expect, considering the higher costs of endovascular grafts, as compared with those for surgical grafts (approximately $6,400 according to literature data). PMID- 11477260 TI - Preoperative evaluation of intracranial aneurysms: usefulness of intraarterial 3D CT angiography and conventional angiography with a combined unit--initial experience. AB - PURPOSE: To assess the usefulness of intraarterial computed tomographic (CT) angiography in conjunction with digital subtraction angiography (DSA) by using a combined CT and angiographic unit in the preoperative evaluation of intracranial aneurysms. MATERIALS AND METHODS: Prospectively, 22 patients with or without subarachnoid hemorrhage underwent CT angiography in conjunction with DSA. Two radiologists independently evaluated DSA and CT angiographic images. Referring neurosurgeons were questioned as to how the additional information provided by CT angiography changed patient treatment. RESULTS: Intraarterial CT angiography was superior to DSA for use in aneurysm detection in three (12%) of 26 aneurysms and for delineation of aneurysm shape, neck, and location in more than half. In 14 (64%) of 22 patients, CT angiography demonstrated 18 additional findings: a very small aneurysm (n = 2), aneurysm shape and neck (n = 6), relationship of the aneurysm to adjacent arteries or bone structure (n = 8), and branches deriving from the aneurysm (n = 2). In four (27%) of 15 patients who underwent surgery or embolization, additional information obtained at CT angiography affected the treatment. CT angiography failed to clearly demonstrate an aneurysm adjacent to bone structures and small perforators, which were derived from the parent artery. CONCLUSION: Intraarterial CT angiography is useful for preoperative evaluation of intracranial aneurysms as a supplement to DSA. PMID- 11477261 TI - Intracranial arterial aneurysms associated with arteriovenous malformations: endovascular treatment. AB - PURPOSE: To evaluate the results of endovascular treatment of patients having both arteriovenous malformations and aneurysms. MATERIALS AND METHODS: Two hundred seventy consecutive patients underwent pretherapeutic selective and superselective angiography. In each patient, the following were recorded: demographic information; clinical symptoms; location of the arteriovenous malformation, including presence of aneurysms; therapeutic interventions and immediate anatomic results; and clinical and angiographic follow-up data. Every patient who had at least one angiographically confirmed arterial aneurysm at presentation was included in the study. RESULTS: Arterial aneurysms were found in 30 (11%) of 270 patients in the population with arteriovenous malformations. Fifteen (50%) of 30 patients with aneurysms had a hemorrhage at presentation. Only 66 (27.5%) of 240 patients without aneurysms had a hemorrhage at presentation. The coexistence of arteriovenous malformations and aneurysms correlated significantly with intracranial hemorrhage at presentation (P <.05). When an aneurysm was believed to be responsible for a hemorrhage and whenever possible, it was treated before the arteriovenous malformation was treated. Treatments were protective against hemorrhage or recurrence of hemorrhage in all cases. Five of 30 patients had neurologic deficits as a result of endovascular treatment of both aneurysms and arteriovenous malformations. CONCLUSION: Findings in this study highlight the importance of recognizing aneurysms in patients with arteriovenous malformations. A strategic focus on the circulatory exclusion of associated aneurysms, especially when such lesions have been responsible for a hemorrhagic episode, is recommended. PMID- 11477262 TI - Quantitative assessment of dural ectasia as a marker for Marfan syndrome. AB - PURPOSE: To establish normal values for lumbosacral dural sac dimensions with magnetic resonance (MR) imaging and to use these values to assess the sensitivity and specificity of dural ectasia as a marker for Marfan syndrome. MATERIALS AND METHODS: MR imaging was performed to measure dural sac diameter (DSD) from L1 through S1 in 44 adult patients with Marfan syndrome and in 44 matched control subjects. DSD values were corrected for vertebral body size, yielding dural sac ratios (DSRs). The control subjects served to establish the upper limit of normal DSR values at the L1 through S1 levels. RESULTS: Cutoff values for normal DSRs for L1 through S1 were 0.64, 0.55, 0.47, 0.48, 0.48, and 0.57. Significant DSR differences were shown at all levels between patients with Marfan syndrome and control subjects (P <.001 at all levels). At L1 through S1, the sensitivity of dural ectasia as a marker for Marfan syndrome was 45%-77%, and the specificity was 95% or greater. By combining levels L3 and S1, dural ectasia as a marker for Marfan syndrome yielded a sensitivity of 95% (42 of 44 patients) and a specificity of 98% (43 of 44). The presence of dural ectasia excelled, compared with the presence of other Marfan syndrome manifestations in the patient population. CONCLUSION: Abnormal DSR values at L3 or S1 can be used to identify Marfan syndrome with 95% sensitivity and 98% specificity. PMID- 11477263 TI - Infrarenal abdominal aortic aneurysms at multi-detector row CT angiography: intravascular enhancement without a timing acquisition. AB - In 70 patients referred for evaluation of aortoiliac aneurysm disease, multi detector row computed tomography was performed with a uniform 25-second delay from the initiation of intravenous administration of a 150-mL bolus of contrast material at 4 mL/sec. In all patients, adequate enhancement (>200 HU) of the aorta and intense enhancement of iliofemoral runoff was achieved without venous contamination. PMID- 11477264 TI - Carotid bifurcation: evaluation of time-resolved three-dimensional contrast enhanced MR angiography. AB - A magnetic resonance (MR) angiographic protocol was evaluated in the carotid bifurcation with use of a pulse sequence for time-resolved three-dimensional imaging of contrast material kinetics. The enhancement ratio, a quantitative measure of contrast enhancement, indicated that all studies included an image obtained near the peak of the intraarterial concentration of contrast agent (enhancement ratio, 90% +/- 9 [standard deviation]). Studies acquired at a higher frame rate (4.1-4.9 seconds) exhibited less venous enhancement (enhancement ratio, 25% +/- 16) than studies acquired with slower (6.0-9.6-second) frame rates (enhancement ratio, 46% +/- 25). PMID- 11477265 TI - Aortic dissection: percutaneous management with a separating stent-graft- preliminary results. AB - The authors used a separating stent-graft to treat Stanford type B aortic dissection. The separating stent-graft consists of two stents: a stent-graft and an inner bare stent. The separating stent-graft has three parts: a proximal stent, a graft made of synthetic polyester textile fiber, and a distal stent. A 12-F introducing sheath was used. After the separating stent-graft was placed, false-lumen thrombosis was evident in all six patients during a follow-up period of 206 days. The major advantages of this technique are that a cutdown and blood pressure control are not required. PMID- 11477266 TI - Theory of high-speed MR imaging of the human heart with the selective line acquisition mode. AB - Selective line acquisition mode (SLAM) reduces magnetic resonance imaging time by a factor n relative to conventional techniques. Seventeen patients with cardiac disease and three volunteers were examined with SLAM and two-frame interpolation (2FI). SLAM images were sharper than 2FI images and showed well-defined endocardial borders. SLAM is best suited for fast imaging of moving objects, such as the heart, confined to 1/n of the field of view. PMID- 11477267 TI - Standardized perfusion value: universal CT contrast enhancement scale that correlates with FDG PET in lung nodules. AB - The standardized enhancement value and standardized perfusion value allow comparison between different methods for quantification of contrast enhancement during computed tomography (CT). Standard perfusion values calculated from CT measurements of perfusion within pulmonary nodules compared favorably with those derived from previously reported enhancement data and correlated with standardized uptake values obtained from positron emission tomographic images (r = 0.8, P <.01). PMID- 11477268 TI - Pneumothorax rate during CT-guided lung biopsies. PMID- 11477269 TI - Complications of radio-frequency thermal ablation. PMID- 11477270 TI - Evaluation of the human limbic lobe. PMID- 11477275 TI - Extra domain-positive fibronectins in arthritis: wolf in sheep's clothing? PMID- 11477276 TI - From bench to bedside: discovering rules for antibody design, and improving serotherapy with monoclonal antibodies. AB - Anti-T-cell monoclonal antibodies (mAbs) form a unique class of therapeutic agent. Their precise specificity offers tremendous potential for the treatment of autoimmune and inflammatory diseases but also prevents meaningful preclinical animal studies. In particular, adverse reactions to therapy may be unanticipated, and the first administration of a novel T-cell mAb to a patient thus marks the beginning of a unique experiment. By comparing clinical parameters and laboratory measurements, small-scale pilot studies can provide detailed information about mAb biology that both predicts and suggests solutions to the complications of therapy. In this essay I illustrate this concept with reference to three specific areas: lymphocyte depletion, mAb immunogenicity and cytokine-release syndromes. In each case, systematic clinical and laboratory science has improved our understanding of the problem and suggested solutions; most of these solutions have been or are being adopted. Thus, small, open studies are an essential step in the development of novel mAbs, provide an ideal platform for the study of mAb biology, and serve as an early warning system for potential adverse effects. PMID- 11477277 TI - Alternatively spliced EDA-containing fibronectin in synovial fluid as a predictor of rheumatoid joint destruction. AB - OBJECTIVES: Fibronectin containing the EDA region (EDA(+)Fn), a molecule important for rheumatoid joint destruction, was measured in relation to the progression of joint destruction in rheumatoid arthritis (RA). METHODS: Total Fn and EDA(+)Fn were measured by ELISA, and the concentrations of Fn in plasma and synovial fluid were compared prospectively for 2 yr with the progression of joint destruction in 41 knee joints of 37 patients with RA. The extent of joint destruction was assessed by the Larsen score and joint space narrowing in X-ray films taken before and 2 yr after measurement of EDA(+)Fn. RESULTS: The concentration of synovial fluid EDA(+)Fn showed a positive correlation with the progression of joint destruction in RA (r=0.78). While total Fn in synovial fluid also showed a correlation with joint destruction (r=0.54), total Fn and EDA(+)Fn in plasma showed no correlation with joint destruction. The concentration of synovial fluid EDA(+)Fn was significantly higher in patients who underwent joint replacement after the measurement of EDA(+)Fn than in those who did not receive surgery (P<0.029). CONCLUSION: Synovial fluid EDA(+)Fn can be a predictor of subsequent joint destruction in RA. PMID- 11477278 TI - Cytokines play an aetiopathogenetic role in fibromyalgia: a hypothesis and pilot study. AB - OBJECTIVE: To measure soluble factors having a possible role in fibromyalgia (FM) and compare the profiles of patients with recent onset of the syndrome with patients with chronic FM. METHODS: The production of cytokines, cytokine-related molecules, and a CXC chemokine, interleukin (IL)-8, was examined. Fifty-six patients with FM (23 with <2 yr and 33 with >2 yr of symptoms) were compared with age- and sex-matched healthy controls. Cytokines and cytokine-related molecules were measured in sera and in supernatants of peripheral blood mononuclear cells (PBMC) that were incubated with and without lectins and phorbol myristate acetate (PMA). RESULTS: No differences between FMS and controls were found by measuring IL-1beta, IL-2, IL-10, serum IL-2 receptor (sIL-2R), interferon gamma (IFN gamma), and tumour necrosis factor alpha (TNF-alpha). Levels of IL-1R antibody (IL-1Ra) and IL-8 were significantly higher in sera, and IL-1Ra and IL-6 were significantly higher in stimulated and unstimulated FM PBMC compared with controls. Serum IL-6 levels were comparable to those in controls, but were elevated in supernatants of in vitro-activated PBMC derived from patients with >2 yr of symptoms. In the presence of PMA, there were additional increases in IL 1Ra, IL-8 and IL-6 over control values. CONCLUSIONS: In patients with FM we found increases over time in serum levels and/or PBMC-stimulated activity of soluble factors whose release is stimulated by substance P. Because IL-8 promotes sympathetic pain and IL-6 induces hyperalgesia, fatigue and depression, it is hypothesized that they may play a role in modulating FM symptoms. PMID- 11477279 TI - Monoclonal antibodies against human ribosomal P proteins penetrate into living cells and cause apoptosis of Jurkat T cells in culture. AB - OBJECTIVE: This study was designed to determine the role of autoantibodies to the ribosomal P protein (anti-P Abs) in the pathogenesis of systemic lupus erythematosus (SLE) using monoclonal anti-P antibodies (anti-P mAbs). METHODS: Anti-P mAbs were prepared by a standard hybridoma procedure using recombinant human P1 and P2 proteins as immunogens. We studied the reactivities of these mAbs to P proteins, their binding and penetration capabilities in different cell lines and their apoptotic effects on Jurkat T cells. RESULTS: In addition to recognizing human P0, P1 and P2 proteins, the anti-P mAb 9B6-4 bound to 20-40% and penetrated 50-90% of astrocytes, Jurkat T cells and lung cancer cells via the P0 surface protein. Treatment with the mAb 9B6-4 also caused increases in the percentages of Jurkat T cells in the sub-G1 phase of the cell cycle (14.8%) and undergoing apoptosis (21.3%). CONCLUSION: Anti-P autoantibodies may play a role in the pathogenesis of lymphopenia or lymphocyte dysfunction in SLE. PMID- 11477280 TI - Evidence for inadequate construct validity of the Disease Repercussions Profile in people with rheumatoid arthritis. AB - OBJECTIVE: To re-evaluate the construct validity of the Disease Repercussions Profile (DRP), a measure of handicap in arthritis populations. METHODS: We used the multitrait-multimethod approach to determine convergent and discriminant validity in a postal survey of randomly selected patients with rheumatoid arthritis who had attended the Wellington Regional Rheumatology Unit since 1988. Respondents (n=142) completed the following self-report instruments: Disease Repercussions Profile, EuroQol EQ-5D, Health Assessment Questionnaire, London Handicap Scale and WHOQOL-BREV. RESULTS AND CONCLUSIONS: The pattern of correlation supported the construct validity of the dichotomous response question (DRP domain affected or not) for the activity and relationships domain, provided ambiguous support for the appearance and emotion domain and no support for the social and financial domains. There was no support for construct validity of any of the domains on the importance rating part of the DRP. We suggest that the DRP be interpreted cautiously in aggregated group data. PMID- 11477281 TI - Churg-Strauss syndrome: outcome and long-term follow-up of 32 patients. AB - OBJECTIVES: To study the clinical spectrum and evolution of Churg-Strauss syndrome in order to assess the clinicopathological features of the disease, the response to treatment and the long-term outcome. METHODS: Thirty-two patients with proven allergic and granulomatous angiitis (Churg-Strauss syndrome) and followed up at a single institution were evaluated. They were recruited between 1977 and 1999 from internal medicine departments. Data were obtained retrospectively from medical files in 15 cases and prospectively, using a standardized form, for the remaining patients. RESULTS: All patients had asthma and hypereosinophilia. The lungs, skin and peripheral nervous system were the organs most frequently involved. Antineutrophil cytoplasmic antibodies with antimyeloperoxidase specificity (MPO-ANCA) were detected in 77.8% of tested patients but they were not useful for monitoring disease activity. Extravascular granulomas were rarely seen in tissue biopsies. Forty per cent of the patients were treated with steroids alone. Immunosuppressive agents were added to the treatment when severe neurological, cardiac or gastrointestinal involvement was present. The outcome and long-term survival were good. Clinical relapse was rare after the first year of therapy. Dysaesthesiae of the distal limbs, neurophatic pain and cardiac failure were the most frequent sequelae. CONCLUSIONS: Churg Strauss syndrome is a rare disorder characterized by hypereosinophilia and systemic vasculitis occurring in patients with asthma and allergic rhinitis. Vasculitis commonly affects the lungs, skin and peripheral nervous system. Outcome and long-term survival is usually good with steroids alone or in combination with immunosuppressive agents. The syndrome has a low mortality rate compared with other systemic vasculitides. PMID- 11477282 TI - Comparison of three active therapies for chronic low back pain: results of a randomized clinical trial with one-year follow-up. AB - OBJECTIVES: To examine the relative efficacy of three active therapies for patients with chronic low back pain. METHODS: One hundred and forty-eight subjects with chronic low back pain were randomized to receive, twice weekly for 3 months, (i) active physiotherapy, (ii) muscle reconditioning on training devices, or (ii) low-impact aerobics. Questionnaires were administered to assess pain intensity, pain frequency and disability before and after therapy and at 6 and 12 months of follow-up. RESULTS: One hundred and thirty-two of the 148 patients (89%) completed the therapy programmes and 127 of the 148 (86%) returned a questionnaire at all four time-points. The three treatments were equally efficacious in significantly reducing pain intensity and frequency for up to 1 yr after therapy. However, the groups differed with respect to the temporal changes in self-rated disability over the study period (P=0.03): all groups showed a similar reduction after therapy, but for the physiotherapy group disability increased again during the first 6 months of follow-up whilst the other two groups showed a further decline. In all groups the values then remained stable up to the 12-month follow-up. The larger group size and minimal infrastructure required for low-impact aerobics rendered it considerably less expensive to administer than the other two programmes. CONCLUSIONS: The introduction of low impact aerobic exercise programmes for patients with chronic low back pain may reduce the enormous costs associated with its treatment. PMID- 11477283 TI - Herbal medicines for the treatment of osteoarthritis: a systematic review. AB - OBJECTIVE: Limitations in the conventional medical management of osteoarthritis indicate a real need for safe and effective treatment of osteoarthritis patients. Herbal medicines may provide a solution to this problem. The aim of this article was to review systematically all randomized controlled trials on the effectiveness of herbal medicines in the treatment of osteoarthritis. METHODS: Computerized literature searches were carried out on five electronic databases. Trial data were extracted in a standardized, predefined manner and assessed independently. RESULTS: Twelve trials and two systematic reviews fulfilled the inclusion criteria. The authors found promising evidence for the effective use of some herbal preparations in the treatment of osteoarthritis. In addition, evidence suggesting that some herbal preparations reduce consumption of non steroidal anti-inflammatory drugs was found. The reviewed herbal medicines appear relatively safe. CONCLUSIONS: Some herbal medicines may offer a much-needed alternative for patients with osteoarthritis. PMID- 11477284 TI - Inhibition of neutrophil responses by cyclosporin A. An insight into molecular mechanisms. AB - OBJECTIVE: Cyclosporin A (CsA) is an effective agent in rheumatoid arthritis (RA), slowing joint damage progression. Its therapeutic effect on T lymphocytes has been studied extensively, but there is little information available about neutrophils, the cells responsible for a substantial proportion of inflammation. A study was performed to investigate the in vitro effects of CsA on neutrophil functions triggered by several agonists and determine whether the drug could counteract the binding of formyl-methionyl-leucyl-phenylalanine (fMLP) to its receptor and/or modulate changes in the intracellular Ca(2+) concentration ([Ca(2+)]i). METHODS: CsA was added to neutrophils 5-50 min before the incubation steps for neutrophil function assays (chemotaxis, superoxide anion production, lysozyme release), calcium measurements and receptor binding experiments. RESULTS: CsA appeared to be particularly effective in lowering chemotaxis, superoxide anion production and lysozyme release induced by different agonists. However, it did not significantly affect either basal or agonist-stimulated neutrophil [Ca(2+)]i and the interaction between fMLP and its receptor. CONCLUSIONS: Because of its in vitro inhibition of neutrophil functions, CsA appears to have considerable potential as an anti-inflammatory drug. Moreover, as it is also a potent immunosuppressive agent, it may reduce the progression of joint damage in RA. More work remains to be done to clarify the molecular mechanism of CsA action on neutrophils. PMID- 11477285 TI - Indirect evidence of intra-articular immunoglobulin G synthesis in patients with Chlamydia trachomatis reactive arthritis. AB - OBJECTIVES: To investigate whether B-cell stimulation occurs in joints of Chlamydia trachomatis reactive arthritis patients by comparing the immunoglobulin G (IgG) anti-C. trachomatis antibody responses in serum and synovial fluid (SF). METHODS: The number and spectrum of C. trachomatis antigens recognized by paired serum and SF samples from 16 patients with C. trachomatis reactive arthritis and 20 patients with other inflammatory arthropathies independent of this bacteria, were studied by immunoblotting. The responses to five different Chlamydia antigens were also determined in enzyme-linked immunosorbent assays. RESULTS: In C. trachomatis reactive arthritis patients, a higher number of C. trachomatis antigens was recognized by SF (17.6+/-5.1) than by serum (11.1+/-6.3) IgG and a higher intensity of SF IgG binding to the outer membrane protein 2 (OMP2) was observed. CONCLUSIONS: These results suggest an intra-articular IgG production and a possible role of some Chlamydia antigens like OMP2 in the pathogenesis of C. trachomatis reactive arthritis. PMID- 11477286 TI - Autoantibodies to a 68/48 kDa protein in chronic fatigue syndrome and primary fibromyalgia: a possible marker for hypersomnia and cognitive disorders. AB - OBJECTIVE: To identify antinuclear antibodies (ANA) specific for chronic fatigue syndrome (CFS), and in related conditions such as fibromyalgia (FM) or psychiatric disorders. METHODS: One hundred and fourteen CFS patients and 125 primary and secondary FM patients were selected based on criteria advocated by the Centers for Disease Control and Prevention and by the American College of Rheumatology, respectively. As controls, healthy subjects and patients with either various psychiatric disorders or diffuse connective tissue diseases were included. Autoantibodies were examined by immunoblot utilizing HeLa cell extracts as the antigen. RESULTS: Autoantibodies to a 68/48 kDa protein were present in 13.2 and 15.6% of patients with CFS and primary FM, respectively. In addition, autoantibodies to a 45 kDa protein were found in 37.1 and 21.6% of the patients with secondary FM and psychiatric disorders, respectively. Meanwhile, these two autoantibodies were not found at all in connective tissue disease patients without FM, nor in healthy subjects (P<0.05). As a group, the anti-68/48 kDa positive CFS patients presented more frequently with hypersomnia (P<0.005), short term amnesia (P<0.07) or difficulty in concentration (P<0.05) than those CFS patients without the antibodies. CONCLUSIONS: The presence of the anti-68/48 kDa protein antibodies in a portion of both CFS and primary FM patients suggests the existence of a common immunological background. These antibodies may find utility as possible markers for a clinicoserological subset of CFS/FM patients with hypersomnia and cognitive complaints. PMID- 11477287 TI - Factors predictive of subsequent deterioration in rheumatoid cervical myelopathy. AB - OBJECTIVE: To identify the features of rheumatoid cervical spine disease associated with deterioration resulting in the need for surgical intervention or death. PATIENTS AND METHODS: Patients with rheumatoid cervical myelopathy who underwent cervical spine magnetic resonance imaging (MRI) between 1991 and 1996 were identified. Patients requiring immediate surgical intervention were excluded. The remainder were divided into two groups. Deterioration group: patients requiring surgical intervention during the follow-up period and deaths resulting from cervical myelopathy. Conservative group: all other patients. Relevant clinical features and radiology reports were extracted retrospectively from the casesheet. RESULTS: The deterioration group comprised 11 patients (12%), median time to deterioration 15 months (range 4-84 months). The conservative group included 82 patients. Initial clinical features did not differ significantly between the two groups. Sixty per cent of those patients with compression or impingement at the atlanto-axial level on first MRI deteriorated over a median of 12 months (range 4-36 months). CONCLUSION: Deterioration is likely if there is evidence of cord compromise at the atlanto-axial level on MRI regardless of initial clinical and plain X-ray features. PMID- 11477288 TI - Leg ulcers in patients with rheumatoid arthritis--a prospective study of aetiology, wound healing and pain reduction after pinch grafting. AB - OBJECTIVE: To study the aetiology of leg ulcers in patients with rheumatoid arthritis (RA) and to study healing and pain relief after pinch grafting. METHODS: Twenty patients with RA and leg ulcers were studied. Diagnosis of the ulcers was founded on the clinical picture and measurements of the ankle-brachial pressure index. To detect vasculitis, skin biopsies were taken for immunohistochemistry and histopathology. Pain severity was assessed pre- and post operatively using a visual analogue scale. RESULTS: Ten of the 20 patients had ulcers with multifactorial aetiology. Fifteen had signs of venous insufficiency, 11 had histopathological evidence of vasculitis, four had reduced arterial circulation and two patients had diabetes. Healing after pinch grafting was found in eight patients, all of whom had an ulcer area less than 15 cm(2). Eleven out of 18 patients had pain reduction after pinch grafting. CONCLUSION: The causation of leg ulcers in patients with RA was found to be multifactorial, with vasculitis and venous insufficiency as the main determinants. Pinch grafting seems to be a good alternative to conservative treatment for minor leg ulcers for these patients, regarding both wound healing and pain relief. PMID- 11477289 TI - Renal type AA amyloidosis associated with rheumatoid arthritis: a cohort study showing improved survival on treatment with pulse cyclophosphamide. AB - OBJECTIVE: To determine the incidence of renal AA amyloidosis and its association with rheumatoid arthritis (RA) in a cohort of all renal biopsies at one referral hospital and to measure the effect of a monthly pulse of cyclophosphamide on renal function and survival in these RA patients. METHOD: All renal biopsies with proven AA amyloidosis from a single pathology unit linked to a major nephrology referral unit in a university hospital were selected retrospectively and RA patients were identified. We studied 6931 renal biopsies. The effect of treatment with and without pulse cyclophosphamide on renal function and survival was studied in these patients. RESULTS: From March 1977 to February 1999, the incidence of AA amyloidosis was 2.4 cases/yr. The incidence and prevalence of the association of AA amyloidosis with RA were 0.68 cases/yr and 0.22% (15/6931) respectively. RA patients treated with cyclophosphamide (n=6) had a lower rate of renal function loss (P=0.013) and a higher median survival (P=0.026) than untreated patients (n=9). During the follow-up period, two out of six treated patients (33%) and all nine untreated patients (100%) died. CONCLUSIONS: AA amyloidosis is a rare complication of RA and complicates the evaluation of treatment. This retrospective study suggests that treatment with cyclophosphamide is able to reduce the incidence of end-stage renal failure and to increase survival. Prospective studies are needed to clarify this issue. PMID- 11477290 TI - The LE cell. PMID- 11477291 TI - Haemophagocytic syndrome in a systemic lupus erythematosus patient with antiphospholipid antibodies. PMID- 11477292 TI - Comment on 'ultrasonic measurement of the thickness of human articular cartilage in situ' by Yao and Seedhom. PMID- 11477293 TI - Azathioprine toxicity mimicking a relapse of Wegener's granulomatosis. PMID- 11477294 TI - Trigeminal sensory neuropathy and rheumatoid arthritis: case study of a rare association. PMID- 11477295 TI - Royal College of General Practitioners and British Society for Rheumatology. A joint initiative for joint problems--how to manage demand. PMID- 11477300 TI - Robin goodfellow. PMID- 11477301 TI - Peripheral or central densitometry: does it matter which technique we use? AB - Over the past decade, bone density scans have assumed an essential role in the diagnosis of osteoporosis. Although dual X-ray absorptiometry (DXA) scans of the central skeleton remain widely used, a variety of different types of equipment for measuring peripheral sites is now available. However, the poor correlation between different types of measurement and a lack of consensus on how results from peripheral sites should be interpreted have proved a barrier to the more widespread use of these devices. These issues prompt the following questions: Which technique best identifies patients at risk of fracture? What approaches to scan interpretation ensure the closest agreement among different methods? Does it matter if different patients are selected for treatment on the basis of different techniques? The relative risk (RR)of fracture derived from prospective studies is a key parameter for comparing the clinical value of different techniques. Recent reports confirm the advantages of hip bone mineral density compared with peripheral measurements for predicting hip fracture risk, although for fractures at other sites the differences are inconclusive. Using receiver operating characteristic curves, we show that the guidelines adopted for scan interpretation are of crucial importance for ensuring that the information provided is used effectively. The closest agreement among different techniques is achieved by setting thresholds for peripheral devices that target either the same percentage of the population or the same percentage of future fracture cases as femur DXA. Different methods select different groups of individuals from the total pool of patients who will later sustain a fracture, with the most successful technique being the one with the largest RR value. The emphasis placed by many studies on validating new techniques by studying their correlation with DXA may lead to the clinical value of peripheral devices being underestimated when the key datum is the RR value inferred from prospective fracture studies. PMID- 11477302 TI - Comparison of quantitative ultrasound and dual X-ray absorptiometry in estrogen treated early postmenopausal women. AB - Identifying individuals at risk of developing osteoporosis is important in order to initiate early treatment. Many new techniques have been proposed as alternatives for DXA-scanning. Some of these alternatives certainly have advantages, but none have so far been demonstrated to predict fractures better, or even to identify individuals at risk of osteoporosis as well as with the standard method. In this study, comprising a group of women from the Danish Osteoporosis Prevention Study, we wished to investigate whether a technique based on quantitative ultrasound (QUS) could identify individuals with low BMC/BMD as measured by dual X-ray absorptiometry (DXA). Furthermore, we wished to test whether the method could detect differences between untreated individuals and those treated with hormone replacement therapy. We found that QUS could detect differences between the treated and untreated groups, but it was unable to identify women with low BMD, although it might be able to identify persons not at risk of osteoporosis. Low QUS values should be followed by a regular DXA measurement to confirm the presence of osteoporosis. PMID- 11477303 TI - Importance of precision in bone density measurements. AB - Bone densitometry, regardless of the specific technique, is not perfectly reproducible even when consistently performed in exact accordance with the manufacturer's recommendations. Precision must be quantified at each densitometry facility in precision studies of the various skeletal sites used for monitoring. The precision, as the root-mean-square standard deviation or root-mean-square coefficient of variation, is then used to determine the change in bone density that constitutes the least significant change and the minimum interval between follow-up measurements. Until precision studies are performed, the least significant change cannot be determined for any level of statistical confidence, making the interpretation of serial studies impossible. PMID- 11477304 TI - Calcaneus ultrasound measurements in a convenience sample of healthy youth. AB - We examined age-related changes in quantitative ultrasound of the calcaneus in 311 healthy males and females ages 6.6-20 yr using the Lunar Achilles ultrasound device. This equipment has been adapted for pediatric use with the provision of shims designed to properly position smaller feet relative to the transducers. Broadband ultrasound attenuation (BUA) (decibels/megahertz), speed of sound (SOS) (meters/second), and stiffness index (SI) (percent) increased across the age range until a plateau was reached at 16-18 yr. BUA increased by 40%, SOS by 4%, and SI by 80% across this age range. There was no gender difference in age related gains. Age, weight, height, and hours of weight-bearing physical activity were all significantly associated with BUA, SOS, and SI. After controlling for age and weight, hours of weight-bearing physical activity showed little to no additional effect on these parameters. Short-term in vivo precision using this device was similar in children to that observed in adults in our laboratory; coefficients of variation for between-day measurements were 1.8, 0.6, and 3.2% for BUA, SOS, and SI, respectively. These data support the feasibility of using the Lunar Achilles in evaluating pediatric bone mass. The ability of this technique to discriminate between osteopenic and normal children remains to be determined. PMID- 11477305 TI - Optical processing of radiographic trabecular pattern versus bone mineral density of proximal femur as measures of bone strength. AB - The purpose of the present study was to evaluate a noninvasive method that utilizes optical processing to analyze the trabecular pattern on bone radiographs. The trabecular network on proximal femur radiographs of 17 intact cadaveric femora was analyzed by optical Fourier transform, yielding a trabecular bone index (TBI) at several locations. The bone mineral density (BMD) of the proximal femur was measured by dual X-ray absorptiometry. Dimensions of the proximal femur were obtained from the radiograph. The bones were fractured in a "fall configuration" to yield the fracture load. A multiple regression model, combining only radiograph- derived parameters-bone dimensions and the TBI at the intertrochanteric region and at the greater trochanter-yielded a correlation of 0.938 with the fracture load. A model combining the BMD at the greater trochanter and at the neck yielded a correlation of 0.928 with the fracture load. When all the variables were introduced into a combined analysis, the correlation with the fracture load was 0.973. The TBI obtained by optical processing of the trabecular bone pattern on femoral radiographs together with bone dimensions derived from these radiographs may serve as an effective estimate for hip fracture risk. PMID- 11477306 TI - Regional bone mass measurement from whole-body dual energy X-ray absorptiometry scan. AB - There are no data on the relative accuracy and precision of regional bone mass measurement from whole-body dual energy X-ray absorptiometry (DXA) scans in small young subjects. Twelve domestic swine piglets (2550-17,660 g) were scanned on a single-beam and on a fan beam densitometer using each humerus and femur as the region of interest to determine the validity of five different scan modes: two infant whole body (IWB), two spine, and one rat whole body (RWB) scan mode in the determination of regional bone mass measurements. DXA bone mineral content (BMC( measurements from RWB and IWB fan beam and IWB single-beam scans were highly predictive of ash weight (adjusted r2 = 0.98, 0.94, 0.94, respectively). Correlation between left and right limbs was highly significant (p < 0.001 for all comparisons) for ash weight (r = 0.99) and for DXA measurements of BMC (r = 0.92-0.99), area (r = 0.92-0.99), and bone mineral density (r = 0.87-0.99) for all modes of DXA scan. Repeatability (as standard deviation of differences of repeated scans) varied with scan mode and DXA parameters and ranged from 1.5 to 7.6%and from 1.8 to 14.7% for intra-and interoperator, respectively. We conclude that regional DXA measurements from IWB and RWB scans can be assessed accurately and with adequate precision for clinical use in subjects with low bone mass comparable with infants and young children. The RWB scan is useful for research studies. However, appropriate training and documentation of precision errors is needed to minimize repeatability errors. PMID- 11477307 TI - Comparison between quantitative calcaneal and tibial ultrasound in a Dutch Caucasian pediatric and adolescent population. AB - In the field of bone densitometry, attention has recently been focused on the pediatric population. Quantitative ultrasound (QUS) as bone assessment technique has many advantages for children in comparison with bone assessment techniques that use ionizing radiation. In this pilot study, we investigated the use of calcaneal and tibial QUS systems in a healthy Caucasian pediatric population. We studied 120 healthy Caucasian Dutch children between ages 7 and 19 yr: 53 boys (mean age of 12.5 yr, range 4.5-18) and 67 girls (mean age of 13.5 yr; range 7.1 19). We recruited children from a large population who previously had participated in a bone assessment study performed at our hospital. Two operators performed calcaneal QUS of the right calcaneus and tibial QUS of the right tibia. The correlation between calcaneal and tibial ultrasound was modest but significant (r = 0.29; p < 0.01). Using the calcaneal device, we found in girls a weak positive correlation between skeletal age and speed of sound (SOS) (r = 0.38), broadband ultrasound attenuation (r = 0.57), and quantitative ultrasound index (r = 0.46), all with a value of p < 0.01. For boys all parameters failed to reach significance. Using the tibial device, we found a good correlation between skeletal age and SOS in girls (r = 0.76) and modest correlation in boys (r = 0.50), both with a value of p < 0.01. This is one of the first studies to present a comparison between two ultrasound techniques in children. At present we feel that, in light of the poor correlation with skeletal age, calcaneal ultrasound has yet to prove its efficacy in children. Tibial ultrasound seems to be a good bone assessment technique in children. PMID- 11477308 TI - Sex differences in bone mass acquisition during growth: the Fels Longitudinal Study. AB - Risk of osteoporosis in later life may be determined during adolescence and young adulthood. The present study used longitudinal data to examine the accumulation of bone mineral content (BMC) and bone mineral density (BMD) in Caucasian subjects ages 6-36 yr. Growth in BMC and BMD (measured by dual X-ray absorptiometry; Lunar, Madison, WI) of 94 males and 92 females was monitored for a mean period of 4.29 yr. The main findings were that there were no sex differences in BMC or BMD during the prepubertal stage; however, females had significantly higher BMD of the pelvis and BMC and BMD of the spine during puberty, and postpubertal males generally had significantly higher BMC and BMD than their female counterparts. In addition, the longitudinal rate of bone accumulation in both sexes increased rapidly during childhood and adolescence and was nearly complete at the end of puberty. Finally, peak BMC and BMD was achieved between the ages of 20 and 25 and occurred earlier in females than in males. The rates of growth and timing of peak bone mass as reported here define the crucial period during which intervention protocols should be developed for maximizing skeletal mass to prevent the development of osteoporosis. PMID- 11477309 TI - Bone mineral density response to long-term bisphosphonate therapy in fibrous dysplasia. AB - Fibrous dysplasia of bone is a rare disease related to a genetic mutation in which bone formation at osseous sites is altered. In the last few years, bisphosphonates have become one of the choice drugs to treat this disease. A 26 yr-old woman presented after 6 wk of spontaneous right leg pain owing to a fissure fracture of the right femoral neck. She reported precocious puberty at the age of 2, with diagnosis of McCune-Albright syndrome. Radioisotope bone scanning, radiographic, biochemical, and densitometric studies were performed. Treatment with bisphosphonates was started because bone turnover biochemical markers were abnormal. Oral olpadronate followed by iv pamidronate substantially decreased bone resorption. Bone mineral density (BMD) of total skeleton and subareas was assessed by dual X-ray absorptiometry (DXA) throughout the 5 yr of treatment. At the end of this period, BMD of the total skeleton had increased 6.2%. However, BMD of the areas most affected by fibrous dysplasia, the legs and pelvis, had increased 12.7 and 11%, respectively. Region of interest analysis of individual bones of the legs performed with the total skeleton scan revealed that BMD of the areas most affected by fibrous dysplasia was lower than that of the less affected contralateral bones. During the first 3 yr, treatment with bisphosphonates substantially increased BMD of the right femur and tibia (22 and 28%, respectively). After that, values seemed to stabilize. DXA evaluation of the total skeleton and its subareas was useful to evaluate the efficacy of bisphosphonate treatment. Moreover, the plateau observed in BMD values after 3 yr of treatment suggests that treatment could have been discontinued when the densitometric values stabilized. PMID- 11477310 TI - First prospective report with the use of quantitative ultrasound (QUS) in children and adolescents. PMID- 11477311 TI - Current issues in therapeutic drug monitoring of mycophenolic acid: report of a roundtable discussion. PMID- 11477312 TI - Perchlorate clinical pharmacology and human health: a review. AB - Potassium perchlorate has been used at various times during the last 50 years to treat hyperthyroidism. Since World War II ammonium perchlorate has been used as a propellant for rockets. In 1997, the assay sensitivity for perchlorate in water was improved from 0.4 mg/L (ppm) to 4 microg/L (ppb). As a result, public water supplies in Southern California were found to contain perchlorate ions in the range of 5 to 8 ppb, and those in Southern Nevada were found to contain 5 to 24 ppb. Research programs have been developed to assess the safety or risk from these exposures and to assist state and regulatory agencies in setting a reasonable safe level for perchlorate in drinking water. This report reviews the evidence on the human health effects of perchlorate exposure. Perchlorate is a competitive inhibitor of iodine uptake. All of its pharmacologic effects at current therapeutic levels or lower are associated with inhibition of the sodium iodide symporter (NIS) on the thyroid follicular cell membrane. A review of the medical and occupational studies has been undertaken to identify perchlorate exposure levels at which thyroid hormone levels may be reduced or thyrotropin levels increased. This exposure level may begin in the 35 to 100 mg/d range. Volunteer studies have been designed to determine the exposure levels at which perchlorate begins to affect iodine uptake in humans. Such effects may begin at levels of approximately 1 mg/d. Environmental studies have assessed the thyroidal health of newborns and adults at current environmental exposures to perchlorate and have concluded that the present levels appear to be safe. Whereas additional studies are underway both in laboratory animals and in the field, it appears that a safe level can be established for perchlorate in water and that regulatory agencies and others are now trying to determine that level. PMID- 11477313 TI - Pharmacokinetics and resistance mutations affect virologic response to ritonavir/saquinavir-containing regimens. AB - The authors assessed the impact of protease and reverse transcription (RT) mutations and individual pharmacokinetic parameters on virologic response to a four-drug regimen including ritonavir/saquinavir. Treatment was given at the start of the study (M0) to 22 HIV-1 protease inhibitor-naive or pretreated patients. Protease and RT genes were sequenced at M0, at the time of virologic failure, or at the end of the follow-up. Plasma ritonavir and saquinavir peak C(max), C(min), and area under the curve (AUC) were determined based on samples taken 0, 1, 2, 3, 4, 6, 8, and 12 hours after administration. HIV-1 RNA decreased to less than 50 copies/mL in 11 patients (group 1). At M0, five of them had no RT mutation and 10 had three or fewer secondary protease mutations with no new mutation during follow-up. Ritonavir and saquinavir pharmacokinetics showed wide interindividual variability. Treatment failed in 11 patients (group 2): 9 had three to eight protease mutations and a mean of 5.8 RT mutations at M0, with emergence of new mutations during follow-up. Pharmacokinetics was similar to those of group 1. The other two patients with virologic failure showed no baseline primary mutation but were the only patients with insufficient saquinavir and ritonavir AUC. The authors showed the complementarity between drug-resistance genotype and individual pharmacokinetics and the potential utility of AUC and Cmax to manage treatment. PMID- 11477314 TI - Identification of N-acetyltransferase 2 and CYP2C19 genotypes for hair, buccal cell swabs, or fingernails compared with blood. AB - Genotyping of polymorphic drug metabolizing enzymes may be useful to estimate the blood concentration, efficacy, and toxicity of drugs before administration. Blood samples are most generally used for genotyping; however, sampling is invasive and complicated by handling and transport. Therefore, the authors developed genotyping methods using nonblood specimens, and then each genotype was compared with that from blood. Healthy Japanese volunteers provided hairs (n = 50), buccal cell swabs (n = 50), and fingernails (n = 30) for N-acetyltransferase 2 and CYP2C19 genotyping. Recovery of genomic DNA from each nonblood specimen was lower than that from 0.5 mL blood. Using a modification of the DNA extraction and polymerase chain reaction amplification method, genotypes were diagnosed without failure, even for those with very low levels of DNA. Both genotypes from these specimens completely matched the genotypes from the blood of the same subject. These nonblood specimens can be convenient, accessible, and economical alternatives to blood as a source of DNA for genotyping. PMID- 11477315 TI - Confirmation of cocaine use during pregnancy: a critical review. PMID- 11477316 TI - Quantitation of immunosuppression by tacrolimus using flow cytometric analysis of interleukin-2 and interferon-gamma inhibition in CD8(-) and CD8(+) peripheral blood T cells. AB - The authors have determined the frequency of intracellular interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) synthesis by T-cell subsets in whole blood (WB) and isolated lymphocytes in 16 transplant recipients treated with tacrolimus and 10 control patients who were not transplant recipients. The authors also determined the impact of varying amounts of red blood cells (RBC) on immunosuppression by tacrolimus. Samples were analyzed by two-color flow cytometry, and the results were expressed as a ratio of whole blood to isolated lymphocytes. In healthy subjects who were not transplant recipients, the frequency of IL-2--producing CD8(-) and CD8(+) cells was higher in WB than in isolated lymphocytes (mean +/- SD of whole blood to lymphocytes ratio: 1.24 +/- 0.5 and 1.67 +/- 0.62, respectively). Adding varying amounts of RBC had no significant impact on IL-2 production by CD8(-) and CD8(+) T cells. Adding tacrolimus (10 ng/mL) to lymphocyte cultures inhibited (90%) IL-2 production in isolated T cells but not in the whole-blood assay. The dose of tacrolimus required for a 50% inhibition of IL-2 release in T cells was 10-fold higher in cultures with RBC than without. Peripheral blood mononuclear cells (PBMC) isolated from tacrolimus-treated whole blood (WB) showed less IL-2 inhibition than did lymphocytes in the WB. The authors also tested cytokine production in WB and PBMCs in 16 transplant recipients and observed various patterns of reactivity. The frequency of IL-2--producing CD8(-) and CD8(+) cells was similar using two different methods in 10 of 16 patients tested. By contrast, in the remaining six patients the authors observed a significant inhibition of IL-2 production in both CD8(-) and CD8(+) T-cell subsets in the whole-blood assay but not in the isolated lymphocytes. The frequency of CD8(-) IFN-gamma--producing cells was significantly lower in 9 of 16 patients, but the same individuals showed no inhibition of their CD8(+) IFN-gamma T cells. The trough levels of tacrolimus did not predict the level of cytokine inhibition in the whole-blood assay in these patients. The authors' results show that the whole-blood assay for cytokine production can be used for monitoring the in vivo effect of tacrolimus in transplant recipients. PMID- 11477317 TI - Importance of the cytochrome P450 2D6 genotype for the drug metabolic interaction between chlorpromazine and haloperidol. AB - The authors studied the interactive effects of the coadministration of haloperidol and chlorpromazine on plasma concentrations of haloperidol and reduced haloperidol. The subjects were 43 Japanese male schizophrenic inpatients who were concomitantly treated with chlorpromazine before or after monotherapy with haloperidol. Coadministration of chlorpromazine produced significant increases in the plasma concentrations of haloperidol (P < 0.01) and reduced haloperidol (P < 0.001) by an average of 28.5% +/- 83.3% and 160.8% +/- 288.9%, respectively. However, there were marked interindividual variations in the interactive effects of chlorpromazine. The authors analyzed the importance of five CYP2D6 genotypes, *1/ *1, *1/ *10, *10/ *10, *1/*5, and *5/*10 on the percentage of change in plasma concentrations of haloperidol and reduced haloperidol. Patients with the CYP2D6*5 allele (n = 4) showed a significantly smaller increase in plasma concentrations of haloperidol (P < 0.05) and a slightly smaller increase in those of reduced haloperidol (P = 0.074) in response to the coadministration of chlorpromazine compared than those with the CYP2D6*1/*1 genotype (n = 8). Those with the CYP2D6*1/*1 genotype (n = 8) showed a trend toward greater increases in plasma concentrations of haloperidol than those with other genotypes (P = 0.087). PMID- 11477318 TI - Effect of grapefruit juice on the pharmacokinetics of losartan and its active metabolite E3174 in healthy volunteers. AB - Grapefruit juice (GJ), a cytochrome P450 (CYP) 3A4 inhibitor, may affect the pharmacokinetics of drugs metabolized through CYP 3A4. Losartan, an angiotensin II antagonist, is converted into its main active metabolite E3174 by CYP 3A4 and CYP 2C9. The effect of GJ on losartan pharmacokinetics was assessed in a randomized crossover trial. Losartan was given to 9 volunteers with and without GJ. Concentrations of losartan and its E3174 metabolite were determined in serum by a high-performance liquid chromatography method (HPLC). Significant differences were observed in some of the pharmacokinetic parameters of losartan and its metabolite E3174 after losartan administration with and without co administered GJ. The lag time (time to drug appearance in serum) of losartan increased significantly with co-administered GJ. The mean residence time (MRT) and half-life (t(1/2)) of the E3174 metabolite were significantly longer and the area under the concentration--time curve (AUC) of the E3174 metabolite was significantly smaller after concomitant GJ administration. The ratio AUC(losartan)/AUC(E3174) was significantly increased after concurrent grapefruit juice intake. The increased lag time of losartan and the increased MRT and t1/2 and decreased AUC of E3174 were considered indicative of simultaneous CYP 3A4 inhibition and P-glycoprotein activation. The significantly increased AUC(losartan)/AUC(E3174) ratio, however, indicates reduced losartan conversion to E3174 by CYP 3A4 metabolism as a result of co-administered GJ. PMID- 11477319 TI - Removal of 10-hydroxycarbazepine by plasmapheresis. AB - Removal of the oxcarbazepine metabolite 10-hydroxycarbazepine (MHD) by plasmapheresis was evaluated during a series of six plasmaphereses of a 13-year old boy with Rasmussen encephalitis. Plasmapheresis was performed after steady state concentrations of MHD had been achieved with a dose of 2550 mg oxcarbazepine daily. The mean amount of MHD removed per plasmapheresis was 78.9 mg (SD: 6.0 mg), representing 3% to 4% of the daily oxcarbazepine dose and approximately 5% to 6% of body stores of MHD. The mean steady-state trough MHD concentration was 33.3 mg/L (SD: 1.8 mg/L), and this was remarkably stable during the entire plasmapheresis period. The serum concentration of MHD was only mildly reduced by the procedure. The areas under the concentration curve of MHD on the first and sixth day of plasmapheresis were 99% and 94%, respectively, of the pre plasmapheresis values. The results are in agreement with studies on other anticonvulsant medications (carbamazepine, valproic acid, phenobarbital, and phenytoin), indicating that minor fractions (2% to 10%) of body stores of these drugs are depleted during plasmapheresis. The authors conclude that it is unnecessary to adjust the oxcarbazepine dosage when performing single-volume plasma exchanges or even multiple exchanges during an extended period. It is further proposed that plasmapheresis is unlikely to be of therapeutic benefit in the treatment of an oxcarbazepine overdose. PMID- 11477320 TI - Antiretrovirals: simultaneous determination of five protease inhibitors and three nonnucleoside transcriptase inhibitors in human plasma by a rapid high performance liquid chromatography--mass spectrometry assay. AB - An analytical technique using liquid chromatography (LC) coupled with electrospray-mass spectrometry (ESI--MS) has been developed for the simultaneous determination of five protease inhibitors (PIs): saquinavir, indinavir, ritonavir, nelfinavir, and amprenavir; and three non-nucleoside reverse transcriptase inhibitors (NNRTIs): nevirapine, delavirdine, and efavirenz, in human plasma. This assay allows the elution and identification of these drugs in a single run (10 minutes) using a linear gradient with water and acetonitrile. The procedure involves liquid--liquid extraction. High-performance liquid chromatography (HPLC) separation was achieved on a C18 reversed-phase column, with a linear gradient elution followed by mass spectrometry detection. The calibration curves, obtained by automatic process peak area integration, show a good linearity in a range of concentrations between 20 and 10,000 ng/mL (40- 10,000 ng/mL for efavirenz). The limit of detection was approximately 10 ng/mL for seven drugs (25 ng/mL for efavirenz). The coefficients of variation (CV) were always less than 15% for both intraday and interday precision for each compound. The recovery of the eight drugs ranged from 88.5% to 100%. This novel LC/ESI--MS assay provides an excellent method for simultaneous quantitative monitoring of different components of the highly active antiretroviral treatments (HAARTs) in patients treated simultaneously with PIs and NNRTIs, and it has been successfully applied to therapeutic drug monitoring and pharmacokinetic studies. PMID- 11477321 TI - Determination of LSD in urine with high-performance liquid chromatography--mass spectrometry. AB - A rapid, specific, and sensitive high-performance liquid chromatography/mass spectrometry method has been developed for routine determination of lysergic acid diethylamide (LSD) in urine. It includes sample purification by extraction into an organic solvent and back-extraction to an acetate buffer, reversed-phase high performance liquid chromatography, and detection with a single quadrupole mass spectrometer equipped with an atmospheric pressure ionization electrospray interface. Trideuterated LSD was used as internal standard. The limit of detection was 0.02 ng/mL and the calibration curve was linear from 0.05 to 10 ng/mL. Within-and between-day coefficients of variation were 3.5% and 4.0% respectively and extraction recovery was 91%. PMID- 11477322 TI - Nelfinavir plasma levels under twice-daily and three-times-daily regimens: high interpatient and low intrapatient variability. AB - Nelfinavir has been recently approved as a twice-daily (BID) dose regimen, but no evaluation of the influence of this regimen change on patients' protease inhibitor exposure has been published. The aim of this study was to compare trough plasma concentrations of nelfinavir obtained under the 1250-mg b.i.d regimen with the levels achieved with the original 750-mg three-times-daily (TID) regimen in 56 HIV-infected patients. Blood samples were obtained at steady state before the morning dose of nelfinavir. Plasma levels were measured by high performance liquid chromatography. Eleven and 45 patients were following TID and BID regimens, respectively. Trough concentrations ranged from 0.14 to 11.74 mg/L and from 0.36 to 10.57 mg/L under TID and BID regimens, respectively. Large interpatient (coefficient of variation: 153%) and modest intrapatient (45%) variabilities of nelfinavir levels were observed. Twenty-one patients (38%) and six patients (11%) had levels above and below, respectively, the trough nelfinavir range (1.0--3.0 mg/L) recommended by the manufacturer. Trough levels are not affected by the dosing regimen; they mainly reflect the important interindividual variability, while remaining fairly stable over time. Many subjects had plasma levels repeatedly outside the assumed therapeutic range. Dose adjustment based on therapeutic drug monitoring may contribute to optimizing antiretroviral therapy. PMID- 11477323 TI - Determination of ketobemidone and its metabolites in plasma and urine using solid phase extraction and liquid chromatography-mass spectrometry. AB - The authors developed a sensitive, specific, and rapid liquid chromatography- mass spectrometry (LC-MS) method for determining ketobemidone and its major metabolites in plasma and urine. The method involves a solid-phase extraction, high-performance liquid chromatography (HPLC), and electrospray mass spectrometry. The limit of quantification for ketobemidone and norketobemidone was 3 nmol/L. Recovery rates for ketobemidone and norketobemidone were 84.8% and 81.1%, respectively. Coefficients of variation (CV) ranged from 2.8 % to 9.5%. The method was used to determine ketobemidone and its major metabolites in clinical samples from relevant patient groups. PMID- 11477324 TI - An automated method for the bioanalysis of vincristine suitable for therapeutic drug monitoring and pharmacokinetic studies in young children. AB - Pharmacokinetic studies in young children require very sensitive methods using low plasma volumes. Although vincristine has been used as an antineoplastic drug for almost 40 years, data on vincristine pharmacokinetics and pharmacodynamics are scarce, especially in young children. One of the reasons for this is the lack of a specific and sensitive assay suitable for small plasma volumes. Therefore the authors aimed to improve an existing high-performance liquid chromatography (HPLC) assay by changing the solid-phase extraction material and by using a more sensitive and controlled electrochemical detector. An on-line solid-phase extraction was used with a preconcentration column of 10 * 3 mm ID containing octadecyl silane (ODS) reversed-phase material and an analytical microsphere C18 column. The mobile phase was unchanged and consisted of 35% phosphate buffer 0.02 mol (pH 7.00 +/- 0.10), 50% methanol, and 15% acetonitrile. Detection was performed with a new electrochemical detector. This detector comprised a highly stable Faraday-shielded oven compartment that accommodated a column and flowcell. The flowcell had a spacer thickness of 0.25 microm set at 830 mV. It also had an excellent signal-to-noise ratio, which resulted in very sensitive electrochemical analysis. These improvements resulted in a lower required sample volume of only 0.3 mL instead of 1.2 mL plasma with a very low limit of quantitation of 0.483 microg/L according to good laboratory practice (GLP) rules. The intraday coefficients of variation were 6.2% (0.483 microg/L) and 4.2% (18.4 microg/L). The interday coefficients of variation were 10.3% (0.483 microg/L) and 8.5% (18.4 microg/L). PMID- 11477325 TI - Fluvoxamine but not sertraline inhibits the metabolism of olanzapine: evidence from a therapeutic drug monitoring service. AB - Therapeutic drug monitoring data of the new atypical neuroleptic drug olanzapine were used to study interactions with the selective serotonin reuptake inhibitors fluvoxamine and sertraline. The distribution of the ratio of concentration/daily dose (C/D; ng/mL per mg/d) of olanzapine was compared in three groups: patients treated with olanzapine (n = 134), patients treated with olanzapine plus fluvoxamine (n = 10) concomitantly, and patients treated with olanzapine plus sertraline (n = 21) concomitantly. No significant difference was seen between the olanzapine and the olanzapine plus sertraline groups. Patients receiving fluvoxamine in addition to olanzapine had C/D ratios that were in the mean 2.3 fold higher than patients receiving olanzapine without additional fluvoxamine. This indicated that fluvoxamine inhibits the metabolism of olanzapine, probably because of inhibition of cytochrome P450 (CYP) 1A2, whereas sertraline is unlikely to interfere with the metabolism of olanzapine. Combination therapy of olanzapine and fluvoxamine should be used cautiously, and therapeutic drug monitoring should be instituted to avoid olanzapine-induced adverse effects or intoxications. PMID- 11477326 TI - Stereoselective pharmacokinetic analysis and antiepileptic activity of N-2 hydroxypropyl valpromide, a central nervous system--active chiral valproylamide. AB - The purpose of this study was to evaluate the anticonvulsant activity and pharmacokinetics (PK) of a novel chiral CNS-active 2-hydroxypropyl valpromide (HP VPD), a derivative of valproic acid (VPA). The individual enantiomers, R, S, and racemic (R,S)-HP-VPD were synthesized and evaluated for their pharmacokinetics and pharmacodynamics in a stereoselective manner. A stereoselective gas chromatography (GC) assay for simultaneous quantification of HP-VPD enantiomers in plasma and urine was developed and used to investigate the pharmacokinetics of HP-VPD in dogs. Pharmacodynamic analysis in rats showed that (S)-HP-VPD was 2.5 times more potent as an anticonvulsant in the maximal electroshock seizure (MES) test than its enantiomer and approximately 10 times more potent than VPA. No significant differences were observed in major PK parameters (clearance, volume of distribution, and half-life) between S and (R)-HP-VPD, and this suggested that pharmacodynamic differences could be attributed to the intrinsic pharmacodynamics of each enantiomer rather than to a preferable pharmacokinetic profile. The pharmacokinetic (metabolic) analysis showed that the fraction metabolized to HP VPD-glucuronide ranged from 5% to 7% and no biotransformation of HP-VPD to VPA and 2-ketopropyl valpromide was observed. This is the first report of significant stereoselectivity in the anticonvulsant activity of a valproylamide with a chiral carbon situated on the alkyl chain of the amine moiety. PMID- 11477327 TI - Antipsychotic radioreceptor assay: a modification identifying selective receptor effects. AB - Radioreceptor assays offer the advantage of a single assay that can assess uniform exposure to multiple chemical compounds. The advent of atypical antipsychotic agents has led to new awareness of the multiple receptor subtypes through which antipsychotic agents may exert their effects, and a renewed interest in comparative drug trials of antipsychotics. The objective of this study was to show the development and validation of antipsychotic radioreceptor assays using clonal cell lines stably expressing isolated human receptors. Model assays were developed using the dopamine(2) (D(2)) and D(4) receptors. D(2) and D(4) activities measured by radioreceptor assay in plasma of antipsychotic treated subjects were highly correlated with high-performance liquid chromatography determinations of antipsychotic concentrations. Similarly, for a variety of typical and atypical antipsychotic agents, the quotients of D(4)/D(2) activity in plasma of antipsychotic-treated subjects were highly correlated with the quotients of D(4)/D(2) affinities of these agents. Valid receptor-selective antipsychotic assays can be established and may have utility for dissecting the in vivo activity of atypical antipsychotics in relation to specific outcomes in clinical trials. PMID- 11477328 TI - The role of acetaldehyde in pregnancy outcome after prenatal alcohol exposure. AB - It is not known why some heavy-drinking women give birth to children with alcohol related birth defects (ARBD) whereas others do not. The objective of this study was to determine whether the frequency of elevated maternal blood acetaldehyde levels among alcoholics is in the range of ARBD among alcoholic women. MEDLINE was searched from 1980 to 2000 using the key words acetaldehyde, pharmacokinetics, and alcoholism for controlled trials reporting blood or breath acetaldehyde levels in alcoholics and nonalcoholics. Separately, using the key words fetal alcohol syndrome, epidemiology, prevalence, incidence, and frequency, articles were identified reporting ARBD incidences among the offspring of heavy drinkers. Of 23 articles reporting acetaldehyde levels in alcoholics, four met the inclusion criteria. Forty-three studies reported on the rate of ARBD in heavy drinkers, and 14 were accepted. Thirty-four percent of heavy drinkers had a child with ARBD, and 43% of chronic alcoholics had high acetaldehyde levels. The similar frequencies of high acetaldehyde levels among alcoholics and the rates of ARBD among alcoholic women provide epidemiologic support to the hypothesis that acetaldehyde may play a major role in the cause of ARBD. PMID- 11477329 TI - Serum levels and cardiovascular effects of tricyclic antidepressants and selective serotonin reuptake inhibitors in depressed patients. AB - Tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs) are used to treat depression. Whereas cardiovascular effects have occasionally been reported during controlled studies with SSRIs, TCA treatment poses a well-known problem in this respect. To investigate the putative correlation between antidepressant dose or serum levels and adverse effects, the authors devised a naturalistic study to evaluate the tricyclic antidepressants' and SSRIs' effect on the cardiovascular system. The authors also compared antidepressant serum levels to adverse effects. Inpatients treated with TCAs or SSRIs were included; an electrocardiogram (ECG) and a Schellong test were carried out on the day patients entered the hospital and during steady-state treatment with antidepressant drugs when blood was drawn for therapeutic drug monitoring. The patient population consisted of 114 acutely depressed patients; 81 patients were treated with TCAs and 33 with SSRIs. The TCAs comprised amitriptyline (n = 43), clomipramine (n = 11), doxepin (n = 19) and imipramine (n = 8); the SSRIs comprised fluvoxamine (n = 14) and paroxetine (n = 19). In TCA-treated patients, the authors observed the same type of abnormalities in conduction and orthostatic hypotension as had been observed earlier. The authors also observed cases of first-degree atrioventricular block, prolonged QTc interval, and orthostatic hypotension in SSRI-treated patients. Thus SSRIs also appear to affect the cardiovascular system, which might pose a problem for patients with preexisting conduction disease. The authors observed a strong correlation between the decrease in systolic pressure and antidepressant serum concentration (except for clomipramine and paroxetine), suggesting that antidepressant serum level is a better correlate than dose. PMID- 11477330 TI - Fluorescence polarization immunoassay: can it result in an overestimation of vancomycin in patients not suffering from renal failure? AB - It has been reported in scientific data that fluorescence polarization immunoassay (FPIA) results in overestimation of vancomycin in patients with renal failure. This overestimation is caused by interference of the degradation product, CDP-1, in this assay. Increases in vancomycin levels have also been reported in patients not suffering from renal failure (nonrenal failure patients) who are receiving vancomycin therapy for approximately 10 days or more. The authors tested whether this increase in vancomycin in nonrenal failure patients is a result of CDP-1 interfering with FPIA or a change in the pharmacokinetics of the drug. Serum vancomycin peak and trough samples were obtained from 10 adult (mean age +/- SD: 55.9 years +/- 17.5) nonrenal failure patients (mean ClCr +/- SD: 76.2 mL/min +/- 29.20) receiving vancomycin therapy for at least 10 days. These peaks and troughs were obtained at steady state and again at approximately 10 days of therapy. All serum samples were analyzed initially by fluorescence polarization immunoassay (FPIA, TDx) (Abbot Diagnostics; Irving, TX) and again by enzyme multiplied immunoassay (EMIT Vancomycin Assay) (Dade Behring; San Jose, CA). Statistical analysis (Wilcoxon signed-rank test) determined that there was no difference between the values obtained from the two assays. This demonstrates that the increase in vancomycin levels is not caused by the accumulation of CDP-1 and may be the result of a change in the pharmacokinetics of the drug. PMID- 11477331 TI - Expedient microdetermination of itraconazole and hydroxyitraconazole in plasma by high-performance liquid chromatography with fluorescence detection. AB - The authors describe a useful and rapid micromethod for the analysis of itraconazole (ITZ) and its active metabolite hydroxyitraconazole (HIT) in human plasma. After a simple deproteinization of 100 microL plasma with acetonitrile, the drug, its metabolite, and an internal standard (IS, ketoconazole) were separated on an 8 mm x 10 cm NovaPak (Waters Associates; Milford, MA) C(18) 4 microm particle-size radial compression cartridge. The compounds of interest were detected using a fluorescence detector with the excitation wavelength set at 260 nm and the emission at 365 nm. The mobile phase consisted of 420 mL water adjusted to a pH of 2.5 with phosphoric acid, 580 mL acetonitrile, and 100 microL triethylamine, which was delivered at a flow rate of 3.0 mL/min. This expedient and rugged method is being used to monitor therapeutic levels in bone marrow transplant recipients who are taking the drug for prophylaxis. PMID- 11477332 TI - Can fluconazole concentrations in saliva be used for therapeutic drug monitoring? AB - The saliva/plasma concentration ratio of fluconazole was investigated in 22 HIV-1 infected individuals with an oropharyngeal Candida infection to determine whether saliva fluconazole concentrations could provide useful information for therapeutic drug monitoring in this population. Steady-state paired plasma and saliva samples were obtained after approximately 1 week of treatment with 50-or 100-mg fluconazole as capsules. A significant correlation between plasma and salivary levels of fluconazole was observed. The median saliva/plasma concentration ratio was 1.3 and was independent of the ingested dose and the plasma fluconazole concentration. The prediction of fluconazole concentrations in plasma from the concentrations in saliva was, although unbiased, not precise. From these findings, the authors conclude that although stimulated salivary fluconazole concentrations are significantly correlated with plasma concentrations, it is not possible to predict plasma fluconazole levels from the salivary concentrations with adequate precision. However, saliva fluconazole concentrations have sufficient value to test for compliance and even semiquantitative prediction of plasma concentrations. PMID- 11477333 TI - Instability of olanzapine in calf serum. PMID- 11477334 TI - Solid organ transplantation in patients with HIV infection. PMID- 11477335 TI - Why do pancreas transplants thrombose? PMID- 11477336 TI - Geranylgeranylacetone, a heat shock protein inducer, prevents primary graft nonfunction in rat liver transplantation. AB - BACKGROUND: Heat shock proteins (HSPs) are well known as cytoprotective proteins. Geranylgeranylacetone (GGA), a nontoxic anti-ulcer drug, was recently shown to have HSP-inducing capacity. In the present study, the activity of GGA was tested in a rat orthotopic liver transplantation (OLT) model to determine whether the compound has beneficial effects in warm ischemia-reperfusion injury. METHODS: Either GGA or a control vehicle was orally administered to donor rats before graft harvest. Harvested livers were subjected to 45-min warm ischemia (37 degrees C) followed by OLT. HSP mRNA expressions and HSP syntheses in the graft livers were evaluated by reverse transcriptase polymerase chain reaction and Western blot analysis, respectively. RESULTS: When the donors were treated with a vehicle, all recipients died of primary nonfunction within 2 days after OLT. In contrast, when the donors were treated with GGA (200 mg/kg per day) for 4 weeks, the 7-day survival rate of recipients was dramatically improved (90%). By giving a high dose of GGA (600 mg/kg per day) for 1 week, a similar improvement in recipient survival was seen (83.3%). GGA administration accumulated mRNA for both HSP72 and HSP90 in the livers even before warm ischemia and facilitated the syntheses of HSP72 and HSP90 after warm ischemia. In addition, GGA pretreatment also significantly reduced the serum levels of tumor necrosis factor-alpha (TNF alpha) after reperfusion. CONCLUSIONS: These findings indicate that both the enhanced induction of HSPs and the suppression of a cytotoxic mediator (TNF alpha) might be involved in the beneficial effects of GGA on ischemia-reperfusion injury. Thus, oral administration of GGA would be a useful tool for preventing primary nonfunction in liver transplantation. PMID- 11477337 TI - Xenotransplantation for brain repair: reduction of porcine donor tissue immunogenicity by treatment with anti-Gal antibodies and complement. AB - BACKGROUND: Transplantation of embryonic neural tissue is a potential treatment for Parkinson's disease. Because human donor material is in short supply, porcine xenografts are considered a useful alternative. Current immunosuppressive therapies fail, however, to protect intracerebral neural xenografts from host CD4 T lymphocytes. To reduce the immunogenicity of porcine donor tissue, we attempted to remove microglial cells with antibodies against the alpha-galactosyl epitope (Galalpha1,3Galbeta1,4GlcNAc-R), or anti-Gal, and complement, and studied whether this pretreatment can reduce direct and indirect T-cell responses to the tissue. METHODS: Brain tissue from 27-day-old pig embryos was dissociated and treated with human anti-Gal and rabbit complement. The microglial content was analyzed by flow cytometry. [3H]thymidine incorporation in cocultures of the brain cells and purified human CD4 T cells was used to determine direct T-cell responses. Indirect T-cell responses were studied by grafting pretreated and control pretreated (no anti-Gal) nigral tissue into the lesioned striatum of immunocompetent rats with 6-hydroxydopamine-induced hemiparkinsonism. Amphetamine induced circling behavior was used to measure graft function. RESULTS: Anti-Gal and complement reduced the microglial content to 11-24% of control and abolished the ability of the brain cells to induce human CD4 T-cell proliferation. Pretreated nigral tissue reduced hemiparkinsonism by more than 50% in five of eight rats at some point during the 10-week follow-up. Rats receiving control pretreated nigral tissue did not display this degree of improvement. CONCLUSIONS: Pretreatment with anti-Gal and complement can reduce the immunogenicity of porcine neural tissue, and might, therefore, be a valuable alternative or supplement to immunosuppression in neural xenotransplantation. PMID- 11477338 TI - A proteasome inhibitor effectively prevents mouse heart allograft rejection. AB - BACKGROUND: We have previously demonstrated in vitro that proteasome inhibitors could suppress proliferation and induce apoptosis of activated T cells. This finding suggests that such inhibitors could be used as a novel category of immunosuppressants in blocking allograft rejection. METHODS: The proteasome inhibitor dipeptide boronic acid (DPBA) was tested in vitro for its inhibitory effect on mouse T-cell proliferation and lymphokine secretion. DPBA was also used in vivo to treat mouse heterotopic heart allograft rejection. Possible side effects of this compound were examined according to blood chemistry of mice treated with DPBA. RESULTS: DPBA suppressed the T-cell proliferation and potently inhibited interleukin (IL)-2, IL-6, IL-10, IL-13, and IFN-gamma produced by anti CD3-activated T cells. Given i.p. starting 1 day after transplantation at 0.66 mg/kg per day for 16 days, or at 1 mg/kg per day for 4 days followed by 0.5 mg/kg per day for 12 days, DPBA could prolong heart allograft survival to 35.5 days (mean survival time, MST) and to 36.2 days, respectively. The control group had MST of 7.3 days. When administrated 72 hr post operation at 1 mg/kg per day for 4 days, DPBA could prolong the graft survival to 19.8 days. During the course of these effective dosages, DPBA had no apparent toxicity in the liver, kidney, pancreas, or heart, according to analysis of blood chemistry. CONCLUSIONS: The proteasome inhibitor could repress allograft rejection in mice without apparent side-effects at the effective dosages. This finding has opened a new dimension in development of novel immunosuppressants for organ transplantation. PMID- 11477339 TI - Desmopressin impairs microcirculation in donor pancreas and early graft function after experimental pancreas transplantation. AB - BACKGROUND: Protective effects of desmopressin in brain dead organ donors oppose reports on a hypercoagulatory potential and an increased leukocyte-endothelial interaction (LEI) after application of the drug. The aim was to evaluate the effect of desmopressin on organ donor's pancreas and early graft function. METHODS: Donor microcirculation was evaluated via intra-vital microscopy (IVM) in 24 BR (di/di) rats with central diabetes insipidus, randomly assigned to groups I (control without desmopressin application), II (single i.v. application, no pretreatment) or group III (single i.v. desmopressin application, s.c. pretreatment for 3 days). Microcirculation in recipients was evaluated 1 hr and 6 hr after syngenic pancreas transplantation. Groups III and I served as organ donors. After IVM specimens were taken for histology and immunohistochemistry. RESULTS: Desmopressin in II vs. I led to temporarily (30') increased LEI (Sticker 274.3+/-87.7 vs. 76.5+/-31.1/mm2 endothelial surface; P<0.01) and impaired microcirculation (MCEV 0.43+/-0.07 vs. 0.99+/-0.06 mm/s; P<0.01). Repeated application reduced MCEV and increased LEI for up to 12 hr. Histology in I vs. III showed increased inflammation (n.s.), necrosis (P<0.05) and vacuolization (P<0.01). Immunohistochemistry revealed increased endothelial P-selectin 20' after application. 6 hr after reperfusion organs from III showed reduced MCEV and increased LEI (P<0.01). CONCLUSION: Repeated application of desmopressin impairs graft microcirculation. Perfusion of the pancreas is significantly reduced at the beginning of organ tissue conservation as well as after reperfusion. These disturbances might partly be due to observed endothelial P-selectin expression. Application of desmopressin up to 12 hr prior to organ explantation may impact graft quality. PMID- 11477340 TI - Treatment of chronic renal allograft rejection in rats with a low-molecular weight heparin (reviparin). AB - BACKGROUND: Low-molecular-weight heparin (LMWH) has been shown to prolong survival of rat cardiac allografts independently from immunosuppressive treatment. Furthermore, long-term treatment reduces the development of chronic graft vascular disease after experimental heart transplantation. The aim of the present study was to determine whether treatment with the LMWH reviparin has a beneficial effect on chronic rejection in a rat renal allograft model. METHODS: Kidneys of Fisher (F344) rats were transplanted into unilaterally nephrectomized Lewis (LEW) recipients. LEW-->LEW isografts served as controls. Animals were treated with cyclosporine (5 mg/kg/d) for the first 10 days. Nephrectomy of the remaining kidney was performed after 10 days. Allografted animals were treated either with reviparin (2 mg/kg/d subcutaneously) for 24 weeks (Allo-24), from week 12 to 24 (Allo-12), or with vehicle for 24 weeks. Proteinuria was determined at regular intervals. Kidneys were harvested after 24 weeks for histomorphological and immunohistochemical evaluation. RESULTS: No major bleeding complications were observed in reviparin-treated animals. Proteinuria was significantly reduced in allografted animals both by early as well as by late onset treatment with reviparin. Transplant glomerulopathy was diminished in Allo 24 and in Allo-12 groups compared to vehicle-treated animals, whereas tubulointerstitial inflammation was influenced only in animals immediately treated with reviparin. Immunohistochemical studies demonstrated a marked reduction of renal monocyte and T-cell infiltration as well as expression of MHC II by treatment with reviparin. CONCLUSIONS: Treatment with the LMWH reviparin significantly improved chronic renal allograft rejection in the F344-to-LEW rat model, both after early and late start of therapy. Although the exact mechanisms of this beneficial effect remain unclear, our data offer a potential new therapeutical approach for prevention of chronic allograft nephropathy. PMID- 11477341 TI - Reduction of severe ischemia/reperfusion injury in rat kidney grafts by a soluble P-selectin glycoprotein ligand. AB - BACKGROUND: Inflammatory leukocyte-endothelium interactions, mediated by selectins, contribute to renal ischemia/reperfusion (I/R) injury. We examined the influence of the soluble P-selectin glycoprotein ligand 1 (sPSGL) on early I/R induced changes in a rat kidney transplantation model with long cold ischemia. METHODS: After 24 hr of cold storage, syngeneic kidneys were grafted into bilaterally nephrectomized rats. Before transplantation, recipients received either 1 mg/kg of sPSGL or vehicle (n=8 per group). Six hours after reperfusion, grafts were removed for light microscopy and immunohistochemistry. Capillary blood flow was measured under a fluorescence microscope by using the concentric circles method. RESULTS: A greater proportion, 74.7+/-7.2% (sPSGL) vs. 28+/-7.4% (controls), of all dye-labeled outer medullary capillaries appeared in the 12 microm radius (P<0.01), indicating dense blood flow, whereas 7.6+/-2.9% vs. 43.3+/-9.7%, respectively, appeared in the 60-microm radius (P<0.05), indicating rarefied blood flow. In the sPSGL-treated group, the extent of severe tubular damage within the inner stripe of the outer medulla was lower compared with controls (37.5+/-8.3% vs. 78.4+/-3.5%, P<0.01). Outer medullary heat shock protein 72 expression was 14.5+/-1.6% in the sPSGL-treated group compared with 9.6+/-1.4% in controls (P<0.05). The number of infiltrating polymorphonuclear leukocytes was similar in both groups. Treatment with sPSGL had no influence on the serum creatinine level. CONCLUSIONS: Our data suggest that impairment of outer medullary blood flow is crucial in I/R injury of kidney grafts with prolonged cold storage. Reduction of capillary blood flow perturbations by sPSGL protects tubular cells from severe structural damage. Blocking early selectin mediated leukocyte adhesion may have therapeutic implications in improving the prognosis of renal transplants with severe I/R injury. PMID- 11477342 TI - The effects of lipid-lowering agents on acute renal allograft rejection. AB - BACKGROUND: Preliminary results from clinical trials suggest that 3-hydroxy-3 methylglutaryl co-enzyme A reductase inhibitors may help prevent acute renal allograft rejection. However, the mechanism for this putative effect of 3-hydroxy 3-methylglutaryl co-enzyme A reductase inhibitors, and whether it is independent of lipid-lowering per SE are unknown. METHODS: Immediately after renal transplantation we randomly allocated (proportioned 2:1:2) patients to: 1) simvastatin (10 mg/day, n=53), 2) simvastatin placebo plus gemfibrozil (dose adjusted for renal function, n=36), and 3) simvastatin placebo (n=52). RESULTS: Simvastatin, but not gemfibrozil, reduced total and low density lipoprotein cholesterol during the first 90 days posttransplant. There were no major adverse effects of therapy. However, there were no effects of treatment on acute rejection. Indeed, survival free of acute rejection at 90 days was 72% in the simvastatin group, 72% in the gemfibrozil group, and 77% in the placebo control group (P=0.771). A post hoc power analysis suggested that there was only a 7.5% chance that a true effect of simvastatin on acute rejection (versus placebo) was not detected, and a 2.5% chance that an effect of gemfibrozil on acute rejection (versus placebo) was not detected in this study. CONCLUSION: Lipid-lowering agents may not reduce the incidence of acute renal allograft rejection. However, additional studies are needed to confirm this observation. In the mean time, many if not most renal transplant recipients should be treated with HMG-CoA reductase inhibitors starting early posttransplant to prevent cardiovascular disease complications. The results of this study suggest that starting lipid-lowering therapy immediately after renal transplantation is both safe and effective in lowering total and low density lipoprotein cholesterol. PMID- 11477343 TI - Influence of pretransplant pregnancy on survival of renal allografts from living donors. AB - BACKGROUND: The presence of a small number of cells of donor origin in organ transplant recipients (microchimerism) may influence allograft survival and may induce tolerance. Postpartum women may be microchimeric to offspring hematopoietic cells up to 27 years. We hypothesized that mothers receiving renal allografts from offspring would have better graft survival compared with either fathers receiving allografts from offspring, or mothers receiving allografts from nonoffspring donors. METHODS: We analyzed 1803 living related kidney transplants from the UNOS database performed between January 1, 1990, and December 31, 1995, for mothers and fathers who received grafts from offspring with one haplotype match. We also compared these mothers with parous females receiving a kidney from nonoffspring donors (spouse and other biologically related or unrelated family members). A multivariate logistic regression method was used to analyze the effect of donor type, as well as other recipient, donor, and transplant characteristics, on graft and patient survival. RESULTS: Mothers receiving one haplotype-matched offspring renal allografts did not have better graft survival at 1 or 3 years posttransplant compared with fathers receiving similar grafts. There was also no difference in graft or patient survival between mothers receiving kidney grafts from either offspring or nonoffspring donors. Graft survival in mothers with multiple pregnancies was poorer than those with a single pregnancy. CONCLUSIONS: It is possible that persistent microchimerism of fetal cells in maternal circulation may, for some mothers, cause a detectable improvement in graft or patient survival. Comparison of female and male recipients from the UNOS database did not reveal any differences in outcomes. If mothers are tolerant to their offspring, our results indicate that this microchimerism may not improve renal allograft or patient survival in offspring donor to maternal recipient combinations. Lastly, more sensitive pretransplant cross-match assays may need to be implemented in multiparous women, given our results. PMID- 11477344 TI - Coagulation markers predicting cardiac transplant rejection. AB - BACKGROUND: Acute cellular rejection in cardiac allografts is a major cause of graft loss, and is associated with activation of the coagulation system. We investigated whether plasma markers of coagulation predict the presence of allograft rejection. METHODS: A total of 132 blood specimens and endomyocardial biopsies were collected from 35 patients, between February of 1997 and May of 1998. We measured plasma prothrombin fragment 1.2 (PF1.2) and p-selectin, fibrinogen, thrombomodulin, and d-dimer. Biopsies were graded according to the International Society of Heart and Lung Transplantation system, with a range of 0 to 4. Grades 0 and 1A were grouped as "no rejection," and the higher grades as "rejection." Linear and logistic regression, accounting for longitudinal data, were the principal analytic tools. RESULTS: p-Selectin level increased progressively with increasing rejection grade (P<0.001). With multivariate analysis, both p-selectin and prothrombin fragment levels significantly predicted rejection. p-Selectin levels were predictive of prothrombin fragment levels (P<0.0001) but not of d-dimer, fibrinogen, or thrombomodulin levels. This model allowed correct prediction of rejection, based on p-selectin and prothrombin fragment values, up to 85% of the time. Dichotomizing patients by a p-selectin level of 65 ng/ml resulted in an odds of rejection of 21.4 [95% C.I. 7.1-64.7] for the patients in the high- compared with the lower risk group. CONCLUSIONS: In heart transplant recipients, p-selectin levels and PF 1.2 levels are highly predictive of organ rejection. The elevation of PF 1.2 suggests that there is systemic generation of thrombin generation. These markers may be useful for noninvasively monitoring patients for organ rejection or for after response to treatment. PMID- 11477345 TI - Liver transplantation for treatment of intrahepatic Osler's disease: first experiences. AB - BACKGROUND: Intrahepatic Osler's disease with multiple arteriovenous malformations and high intrahepatic shunting may lead to secondary pulmonary hypertension followed by right-heart stress and insufficiency. Until now, therapy with arterial embolization, banding, or ligation of the hepatic arteries is still limited and provides unsatisfactory long-term results. Liver transplantation offers another therapeutic option. METHODS: We report on four patients with intrahepatic involvement of Osler's disease who were liver transplanted between 1995 and 1999. All patients suffered from restricted liver function and right heart insufficiency with multiple cardiac decompensations. One patient received one course of embolization, and another received six courses of embolization and then banding of the main hepatic artery before transplantation. In both patients, the clinical symptoms improved for only a few months. RESULTS: All patients had high degrees of intrahepatic arteriovenous shunting, and cardiac output measurements were between 8.0 to 13.3 L/min preoperatively. Preoperative mean pulmonary artery pressure was between 24 to 35 mmHg. After liver transplantation, cardiac output and right-heart diameter decreased or normalized and pulmonary pressure reached the normal range after 2 months. All patients received tacrolimus and steroids for primary immunosuppression. In one case, temporary hemodialysis was necessary for 2 weeks after transplantation, but renal function recovered completely. After follow-up time of 12 to 65 months, all patients had normal graft function and good cardiopulmonary condition. CONCLUSIONS: Indication for liver transplantation should be considered in patients with intrahepatic Osler's disease, high arteriovenous shunting with right-heart stress, and restricted liver function before irreversible fixed pulmonary hypertension leads to severe right-heart insufficiency or failure. Our therapeutic regimen of early liver transplantation in the case of intrahepatic Osler's disease in four patients has promising results. PMID- 11477346 TI - Hepatitis C antibody status and outcomes in renal transplant recipients. AB - BACKGROUND: Hepatitis occurs frequently in patients with end-stage renal disease. In 1997, 0.7% of patients receiving a renal transplant were positive for hepatitis C antibodies. Concern has been raised as to whether these patients are at an increased mortality risk after renal transplantation compared with patients who are hepatitis C antibody negative. To help answer this question, we analyzed data from the United States Renal Data System from October of 1988 through June of 1998. METHODS: Primary study endpoints were patient death and death censored graft loss. Secondary study endpoints included cardiovascular, infectious, malignant, and infection-related death. Kaplan-Meier survival estimates as well as Cox proportional hazard models were used to evaluate the impact of hepatitis C antibody status on the study endpoints. RESULTS: A total of 73,707 patients were analyzed. Patient survival by Kaplan-Meier analysis was higher in hepatitis C positive patients, whereas death censored graft survival trended lower in the very long term. By the Cox model, hepatitis C-positive adjusted patient survival is slightly superior to that of hepatitis C-negative patients. CONCLUSIONS: Renal transplant recipients who are hepatitis C antibody positive do not have an increased risk of death after transplantation compared with hepatitis C-negative recipients. The current policy of transplanting hepatitis C-positive patients without active liver disease seems to incur no excess mortality risk. PMID- 11477347 TI - Randomized trial of tacrolimus plus mycophenolate mofetil or azathioprine versus cyclosporine oral solution (modified) plus mycophenolate mofetil after cadaveric kidney transplantation: results at 2 years. AB - BACKGROUND: A previous report described the 1-year results of a prospective, randomized trial designed to investigate the optimal combination of immunosuppressants in kidney transplantation. Recipients of first cadaveric kidney allografts were treated with tacrolimus+mycophenolate mofetil (MMF), cyclosporine oral solution (modified) (CsA)+MMF, or tacrolimus+azathioprine (AZA). Results at 1 year revealed that optimal efficacy and safety were achieved with a regimen containing tacrolimus+MMF. The present report describes results at 2 years. METHODS: Two hundred twenty-three recipients of first cadaveric kidney allografts were randomized to receive tacrolimus+MMF, CsA+MMF, or tacrolimus+AZA. All regimens contained corticosteroids, and antibody induction was used only in patients who experienced delayed graft function. Patients were followed up for 2 years. RESULTS: The results at 2 years corroborate and extend the findings of the previous report. Patients randomized to either treatment arm containing tacrolimus experienced improved kidney function. New-onset insulin dependence remained in four, three, and four patients in the tacrolimus+MMF, CsA+MMF, and tacrolimus+AZA treatment arms, respectively. Furthermore, patients with delayed graft function/acute tubular necrosis who were treated with tacrolimus+MMF experienced a 23% increase in allograft survival compared with patients receiving CsA+MMF (P=0.06). Patients randomized to tacrolimus+MMF received significantly lower doses of MMF compared with those administered CsA+MMF. CONCLUSIONS: All three immunosuppressive regi-mens provided excellent safety and efficacy. How ever, the best results overall were achieved with tacrolimus+MMF. The combination may provide particular benefit to kidney allograft recipients who develop delayed graft function/acute tubular necrosis. Renal function at 2 years was better in the tacrolimus treatment groups compared with the CsA group. PMID- 11477348 TI - Cyclosporine microemulsion- and mycophenolate mofetil-related lymphoid aggregates are not associated with acute rejection. AB - BACKGROUND: Microemulsion cyclosporine, mycophenolate mofetil, and prednisone have become a common immunosuppressive protocol in renal transplantation. We identified lymphocytic infiltrates in transplant fine-needle aspirates and core biopsies from patients on this regimen without acute rejection clinically or by standardized morphological criteria and investigated this inflammatory response. METHODS: Twenty-eight aspirates from 21 patients were included and assessed in the standard fashion. Nine core biopsies showing interstitial lymphocytic infiltration were evaluated with antibodies against CD3, CD4, CD8, CD20, CD30, CD56, KP1, and epithelial membrane antigen (EMA). Aspirates and biopsies were assessed for tubular cell major histocompatibility complex (MHC) class II antigen and for gamma-interferon (gamma-IFN), interleukin-4 (IL-4), and IL-10 mRNAs by reverse transcription-polymerase chain reaction. RESULTS: Fifteen aspirates showed immune activation solely due to mature lymphocytes and monocytes; 13 had no immune activation. All aspirates were negative for MHC class II antigens. Of 6 activated aspirates assessed for gamma-IFN mRNA, 5 were negative. All 21 patients had similar clinical characteristics and recovered renal function without rejection treatment. The core biopsies had lymphocytes in 5-30% of the interstitium. The cells were 70-85% CD3+, with 50-85% CD4+, 3-10% KP1+, and rare cells CD56+. No T-cell activation was present (EMA- and CD30-). Seven biopsies were assessed and were negative for gamma-IFN mRNA; only one biopsy had weakly positive MHC class II staining. Two activated aspirates were negative for IL-4 and IL-10 mRNA, while three biopsies each contained IL-4 and IL-10 mRNAs. CONCLUSIONS: Inactive interstitial lymphoid infiltrates are frequent in patients on this drug regimen and should not be interpreted as acute rejection, particularly in aspirate samples. These lymphocytes may play a role in long-term allograft acceptance. PMID- 11477349 TI - Risk factors for renal allograft survival from pediatric cadaver donors: an analysis of united network for organ sharing data. AB - BACKGROUND: The shortage of cadaveric donors for kidney transplantation has prompted many centers to use cadaver kidneys from pediatric donors. Use of kidneys from pediatric donors has been shown to have a lower graft survival. METHODS: Recipients receiving cadaver kidneys from pediatric and adult donors between 1988 and 1995 were analyzed. The data were obtained from United Network of Organ Sharing database. The actuarial kidney transplant graft survival was estimated by the Kaplan-Meier method. A logistic regression analysis was used to identify various risk factors for 1-year graft failure. Odds ratios (OR) were estimated for various risk factors. RESULTS: Kidney transplant survival rates for donor age <18 years (n=12,838) at 1, 2, 3, 4, and 5 years were 81.5%, 76.3%, 71.3%, 66.4%, and 61.7%, respectively. The corresponding results for adult donors from age 18 to 50 years (n=35, 442) were 83.5%, 78.4%, 73.1%, 67.9%, and 62.4%, respectively, Log-rank test P<0.01. Pediatric donors were further divided into three groups according to donor age: group I (0-5 years), group II (6-11 years), and group III (12-17 years). The actuarial survival rates for 1, 3, and 5 years for group I (n=2198) were 73.6%, 63.3%, and 55.6%, respectively. The corresponding values for group II (n=2873) were 78.0%, 67.5%, and 57.8% and for group III (n=7767) were 85%, 75.0%, and 64.8%, respectively, P<0.01. Although the recipients of group I had lower graft survival, en bloc grafts (n=751) had much better 1-, 3-, and 5-year graft survival rates (76.3%, 67.7%, and 60.7%, respectively) compared with single grafts (n=1447; 72.2%, 61.1%, and 53.2%, P=0.02) from donors 0 to 5 years. Graft thrombosis as a cause of graft failure was seen in 10% of group I compared with 6% in group II and 5% in group III. In group I, lower OR were seen when an en bloc transplant was performed (0.688, P<0.01) and when donor body weight was>15 kg (0.547, P<0.01). However, OR were elevated in recipients of previous transplants (1.556, P<0.01), with prolonged cold ischemic time (1.097, P=0.03), for black recipients (1.288, P=0.03), and for recipients with body mass index> or =25 (1.286, P=0.02). Progressive increase in the donor age was associated with lower OR in group II (0.894, P<0.01). CONCLUSIONS: (1) Overall, poorer graft survival was seen in pediatric donor transplants, (2) transplant kidney survival with en bloc kidneys was better than a single kidney from donors 0-5 years, (3) progressive increase in donor age was associated with improved graft survival when the donors were 6-11 years, whereas progressive increase in donor weight was associated with improved graft survival when the donors were 0-5 years. PMID- 11477350 TI - Effect of poverty and other socioeconomic variables on renal allograft survival. AB - BACKGROUND: Socioeconomic variables including low income and noncompliance impact negatively upon long-term renal allograft survival, especially in African Americans. We sought to determine whether other socioeconomic variables contributed to noncompliance and allograft survival. METHODS: A detailed history of socioeconomic variables was made at the time of renal transplant evaluation in 450 consecutive candidates, 128 of whom (89 African American, 39 Caucasian) have thus far undergone transplantation. Variables evaluated included household income, zip code income, insurance coverage, years of education, literacy, marital status, pretransplantation compliance, and history of substance abuse as well as the usual pre- and posttransplantation demographics. RESULTS: Immunologic graft loss occurred primarily in young African Americans with income below the federal poverty level, whereas nonimmunologic graft loss was distributed across racial, income, and other socioeconomic variables. Immunologic graft loss was also associated with a greater number of HLA mismatches, lower levels of education, and noncompliance with transplant medications and follow-up visits. Recipients with gross illiteracy, however, had excellent graft survival. Pretransplantation substance abuse, but not pretransplantation compliance, was predictive of posttransplantation noncompliance. By multivariate analysis, posttransplantation compliance emerged as the single most important factor predictive of graft survival. CONCLUSIONS: Immunologic graft loss in our population is related to noncompliance with transplant medications, which occurred primarily in recipients with a pretransplantation history of substance abuse and is not related to an inability to pay for medications at the time of graft loss. A change in criteria for acceptance of transplant candidates with a prior history of substance abuse might significantly improve graft survival in this patient population. PMID- 11477351 TI - Posttransplant immune hepatitis in pediatric liver transplant recipients: incidence and maintenance therapy with azathioprine. AB - BACKGROUND: Cases of so-called autoimmune hepatitis (AH) have been reported after liver transplantation. Our aim was to evaluate the incidence in a series of 471 pediatric liver transplant recipients. METHODS: Between 1984 and 1998, 471 children had orthotopic liver transplantation (OLT). Children are followed up on a regular basis, with full clinical, biochemical, and histologic evaluation at 6 months, 1, 2, 5, 7, and 10 years after OLT. Children with unexplained abnormal liver tests were screened for autoimmune markers (total gamma-globulins, smooth muscle antibodies [SMA], liver kidney microsome antibodies [LKM], antinuclear factor [ANA]). From January of 1998 until December of 1998, autoimmune markers were prospectively searched in all children admitted for regular posttransplant follow-up (n = 118). RESULTS: Eleven of 471 children (2.35%) were found with autoimmune hepatitis, 9 retrospectively and 2 prospectively. None had previous autoimmune liver disease. Patients had a history of steroid-dependent hepatitis. Histology showed variable degree of portal and lobular inflammation, piecemeal necrosis, and bridging collapse. Mean (+/-SDS) aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities at diagnosis were 173+/-145 and 196+/-157 IU/L, respectively (nl<32). Median gamma-globulin levels reached 1365 mg/dl versus 931 mg/dl in controls (P<0.05). Nine had ANA (titer 1/80 up to 1/10,000), 1 SMA (1/320), and 2 LKM1 antibodies (1/1280). Patients did not respond to increasing charge of cyclosporine (n=10) or tacrolimus (n=1). Eleven received steroids (prednisolone: 2 mg/kg per day, then tapered) and azathioprine (1.5 to 2.5 mg/kg per day). All patients normalized within 3 months (mean AST/ALT levels of 26+/-8 and 30+/-9 IU/L). Three had mild to moderate relapse with increase of ALT thereafter. Gamma-globulins decreased to 1190 mg/dl (ns). Amongst the 116 remaining prospectively evaluated patients, 85 had normal evaluation, despite low titers of autoantibodies in 15 (SMA< or =1/40, ANA 1/80). Thirty-one patients had graft dysfunction, related to well-explained posttransplant causes, among which 7 had similar low levels of autoantibodies. CONCLUSIONS: A total of 2.35% of our transplant children present evidence of immune hepatitis after transplantation. Patients do not respond to increasing cyclosporine or tacrolimus levels and require steroid and azathioprine. In view of this specific treatment, systematic screening for "autoimmune" markers is advised in children with liver transplant. PMID- 11477352 TI - Safety and efficacy of hepatitis A vaccination in liver transplantation recipients. AB - OBJECTIVE: Vaccination against hepatitis A (HAV) has been shown to be safe and effective in healthy subjects and in patients with chronic liver disease (CLD). The safety and efficacy of HAV vaccines in liver transplant (OLT) recipients have not been established. The objective of this study is to assess the safety and efficacy of inactivated hepatitis A vaccine in OLT recipients. METHODS: Thirty seven HAV seronegative OLT recipients were enrolled. Patients received two doses of vaccine 6 months apart. Postvaccination IgG anti-HAV were determined at 1, 6, and 7 months after the first vaccine dose. Side effects were monitored for 3 days after each vaccination shot. An unvaccinated control group (45 patients) was followed for evidence of seroconversion. Seroconversion rate was also compared with those reported in healthy patients and in patients with chronic liver disease. RESULTS: Testing was available for all the cases at 1 month, and for 26 and 23 patients at 6 and 7 months, respectively. Only 3 of 37 patients (8%) had seroconversion at 1 month. At 6- and 7-month time points, 5 of 26 (19%) and 6 of the 23 (26%) patients had seroconversion, respectively. Vaccine responders had higher total white blood cell count and lymphocyte count and were further out from transplant compared with nonresponders. None of the unvaccinated patients had seroconversion over the follow-up time. Seroconversion rates in OLT recipients were significantly lower than that reported in healthy individuals (P=0.001) or in pre-OLT patients with CLD (P=0.001). All patients tolerated the vaccine well. CONCLUSIONS: HAV vaccination is safe in OLT recipient. Efficacy of HAV vaccination in OLT recipients, as measured by a commercially available enzyme immunoassay, is low and alternative strategies should be developed to improve response rate. PMID- 11477353 TI - Pretransplant cachexia and morbid obesity are predictors of increased mortality after heart transplantation. AB - BACKGROUND: Extremes in body weight are a relative contraindication to cardiac transplantation. METHODS: We retrospectively reviewed 474 consecutive adult patients (377 male, 97 female, mean age 50.3+/-12.2 years), who received 444 primary and 30 heart retransplants between January of 1992 and January of 1999. Of these, 68 cachectic (body mass index [BMI]<20 kg/m2), 113 overweight (BMI=>27 30 kg/m2), and 55 morbidly obese (BMI>30 kg/m2) patients were compared with 238 normal-weight recipients (BMI=20-27 kg/m2). We evaluated the influence of pretransplant BMI on morbidity and mortality after cardiac transplantation. Kaplan-Meier survival distribution and Cox proportional hazards model were used for statistical analyses. RESULTS: Morbidly obese as well as cachectic recipients demonstrated nearly twice the 5-year mortality of normal-weight or overweight recipients (53% vs. 27%, respectively, P=0.001). An increase in mortality was seen at 30 days for morbidly obese and cachectic recipients (12.7% and 17.7%, respectively) versus a 30-day mortality rate of 7.6% in normal-weight recipients. Morbidly obese recipients experienced a shorter time to high-grade acute rejection (P=0.004) as well as an increased annual high-grade rejection frequency when compared with normal-weight recipients (P=0.001). By multivariable analysis, the incidence of transplant-related coronary artery disease (TCAD) was not increased in morbidly obese patients but cachectic patients had a significantly lower incidence of TCAD (P=0.05). Cachectic patients receiving oversized donor hearts had a significantly higher postoperative mortality (P=0.02). CONCLUSIONS: The risks of cardiac transplantation are increased in both morbidly obese and cachectic patients compared with normal-weight recipients. However, the results of cardiac transplantation in overweight patients is comparable to that in normal weight patients. Recipient size should be kept in mind while selecting patients and the use of oversized donors in cachectic recipients should be avoided. PMID- 11477354 TI - Randomized controlled trial of hand-assisted laparoscopic versus open surgical live donor nephrectomy. AB - BACKGROUND: Laparoscopic live donor nephrectomy for renal transplantation is being performed in increasing numbers with the goals of broadening organ supply while minimizing pain and duration of convalescence for donors. Relative advantages in terms of recovery provided by laparoscopy over standard open surgery have not been rigorously assessed. We hypothesized that laparoscopic as compared with open surgical live donor nephrectomy provides briefer, less intense, and more complete convalescence. METHODS: Of 105 volunteer, adult, potential living-renal donors interested in the laparoscopic approach, 70 were randomly assigned to undergo either hand-assisted laparoscopic or open surgical live donor nephrectomy at a single referral center. Objective data and subjective recovery information obtained with telephone interviews and validated questionnaires administered 2 weeks, 6 weeks, and 6-12 months postoperatively were compared between the 23 laparoscopic and 27 open surgical patients. RESULTS: There was 47% less analgesic use (P=0.004), 35% shorter hospital stay (P=0.0001), 33% more rapid return to nonstrenuous activity (P=0.006), 23% sooner return to work (P=0.037), and 73% less pain 6 weeks postoperatively (P=0.004) in the laparoscopy group. Laparoscopic patients experienced complete recovery sooner (P=0.032) and had fewer long-term residual effects (P=0.0015). CONCLUSIONS: Laparoscopic donor nephrectomy is associated with a briefer, less intense, and more complete convalescence compared with the open surgical approach. PMID- 11477355 TI - Microcirculatory changes in right lobe grafts in living-donor liver transplantation: a near-infrared spectrometry study. AB - BACKGROUND: A continuing shortage of cadaveric liver even for adult patients has motivated not a few centers to proceed to living-donor liver transplantation using right lobe grafts. One of controversies is potential congestion in the graft anterior segment by the deprivation of the middle hepatic vein. METHODS: Hepatic tissue oxygenation and hemoglobin concentration were investigated with a near-infrared spectroscopy in the course of harvesting and implantation in living donor liver transplantation. Twenty adult recipients of right lobe graft were involved in the study. The aim of the analysis was to detect tissue congestion or ischemia. RESULTS: No significant change in mean hepatic tissue oxygenation and hemoglobin was noted in the right lobe during donor operation even after hepatic parenchymal transection, although some trend for relative congestion, i.e., increased tissue hemoglobin, compared with the left lobe was observed. After graft reperfusion in the recipient, both mean hepatic tissue oxygen saturation and hemoglobin decreased significantly in the anterior segment, which was accompanied by increased heterogeneity of tissue hemoglobin and oxygenation. Increased heterogeneity of oxygenation and decreased tissue hemoglobin were observed also in the posterior segment. CONCLUSIONS: The anterior segment in right lobe living-donor liver transplantation is sensitive to ischemia, rather than congestion, at least in the immediate phase after graft reperfusion. The anterior segment seems to be also more prone to circulatory disturbance than the other part of the graft. PMID- 11477356 TI - Variant transthyretin in blood circulation can transverse the blood-cerebrospinal barrier: qualitative analyses of transthyretin metabolism in sequential liver transplantation. AB - BACKGROUND: Although the choroid6 plexus of the brain is one of the most important production sites of transthyretin (TTR), the metabolism of TTR secreted in cerebrospinal fluid (CSF) remains to be elucidated. METHODS: To perform qualitative analysis of variant TTR in CSF of patients who underwent a sequential liver transplantation using an explanted familial amyloidotic polyneuropathy (FAP) ATTR Val30 Met patient's liver, levels and forms of TTR of the two patients were analyzed by means of enzyme linked immunosorbent assay (ELISA) and matrix assisted laser desorption/time-of-flight mass spectrometer (MALDI/TOF-MS), respectively. RESULTS: After the operation, variant TTR levels in serum increased, and in CSF, a significant peak of free form of ATTR Val30 Met was detected in the transplanted patients whose CSF had shown no variant TTR before the operation. CONCLUSIONS: These findings suggest that the variant TTR can cross the blood-CSF barrier and migrate into CSF from blood circulation. Because leptomeningeal amyloidosis occurs in FAP ATTR Val30 Met as the progression of the disease, this information suggests that in addition to peripheral neuropathy, disorders of the central nervous system (CNS) should be given an attention in patients who underwent sequential liver transplantation using an explanted FAP ATTR Val30 Met patient's liver. PMID- 11477357 TI - Optimal alveolar oxygen concentration for cold storage of the lung. AB - BACKGROUND: Ischemia of the lung is different from that of solid organs because the lung contains gas in the alveoli. However, the optimal gas composition in the alveoli during cold storage remains uncertain. We investigated the relationship between the alveolar oxygen concentration and reperfusion injury. METHODS: The lungs inflated with 0% O2, 5% O2, room air, 50% O2, or 100% O2 were reperfused after 8 hR storage at 4 degrees C and pulmonary functions were measured for 120 min using an ex vivo rat lung model. The levels of high-energy phosphate and lipid peroxidation of the lung were analyzed after a PA flush, preservation, and reperfusion. Additionally, respiration of the mitochondria in the lungs was measured after preservation. RESULTS: The pulmonary functions were significantly superior in the 5% O2 group than those in the 0% O2, 50% O2, and 100% O2 groups. Pulmonary edema developed in the 0% O2, 50% O2, and 100% O2 groups, but not in the 5% O2 group. After preservation, the energy level in the lungs decreased only in the 0% O2 group. Although lipid peroxidation of the lungs did not increase in any group after preservation, significant increases were observed in the room air, 50% O2 and 100% O2 groups after reperfusion. State 3 and 4 ratios of the mitochondrial respiration significantly decreased in the lungs of the room air, 50% O2 and 100% O2 groups. CONCLUSIONS: Although the cold-preserved lungs require oxygen, hyperoxygenation induced mitochondrial dysfunction and increased lipid peroxidation and led to deleterious lung function after reperfusion. Therefore, hypoxic conditions that can maintain the energy level of the lung during cold storage would be optimal. PMID- 11477358 TI - Ligand density determines the efficiency of negative selection in the thymus. AB - To study the influence of antigen density on the efficiency of negative selection in the thymus, MHC class I (H-2K(b), K(b)) transgenic mice were generated, which expressed a K(b) transgene under the control of its natural promoter at 33% (K(b lo)) or 150% (K(b-hi)) the surface density of Kb in C57BL/6 (B6, H-2(b)) mice. These mice were crossed to anti-K(b) T-cell receptor (Des-TCR) transgenic mice. In Des-TCRxK(b-hi) double transgenic mice, Des-TCR bearing T cells were completely eliminated during thymocyte maturation. In contrast, in Des-TCRxK(b lo) double transgenic mice, two populations of Des-TCR T cells were evident, which either expressed the Des-TCR at intermediate density in the absence of CD8 (Des-TCR(int)CD8(-)) or expressed both the Des-TCR and CD8 at low density (Des TCRloCD8lo). In the thymus of both types of double transgenic mice, no Des TCR(+)CD4(+)CD8(+) thymocytes were detected, suggesting that deletion of Des-TCR cells occurred before the CD4(+)CD8(+) stage. Because only very few Des-TCR(+) thymocytes were found in Des-TCRxK(b-hi) transgenic mice, deletion of these T cells apparently occurred upon expression of the Des-TCR. By contrast, Des TCRxK(b-lo) transgenic mice showed distinct populations of Des-TCR(int)CD4-8- and Des-TCR(lo)CD8(lo) thymocytes, suggesting that expression of the CD8 coreceptor was required to allow negative selection to proceed. Functional analyses showed that sublethally irradiated Des-TCRxK(b-lo) double transgenic mice were protected from lethal graft-versus-host disease by injected Des-TCR lymph node cells. PMID- 11477359 TI - Expression of complement regulatory proteins on islets of Langerhans: a comparison between human islets and islets isolated from normal and hDAF transgenic pigs. AB - BACKGROUND: The expression of regulators of complement activity (RCAs) on islet cells may be of great importance for protecting them against complement-mediated lysis in the immediate posttransplant period after intraportal islet transplantation. We examined porcine and human islet cells for expression of RCA. We also examined to what extent human decay accelerating factor (hDAF) is expressed on adult and fetal islet cells isolated from hDAF transgenic (TG) pigs having a high transgene expression on endothelial cells. Moreover, the susceptibility of the various types of cells to lysis in human serum and blood was investigated. METHODS: Adult human islets (n=5), normal adult and fetal porcine islets (n=9 and n=8, respectively), and islets from adult and fetal hDAF TG pigs (n=5 and n=6, respectively) were examined. With islet single-cell suspensions and flow cytometry, adult human islet cells were examined for expression of hDAF (CD55), hCD59, and human membrane cofactor protein (hMCP; CD46), while porcine islet cells were examined for expression of pCD59 and pMCP. Islet cells from hDAF TG pigs were also examined for hDAF expression. Porcine peripheral blood lymphocytes, normal and hDAF TG porcine endothelial cell lines, a human endothelial cell line, and the human cell line U937 served as controls. Islet cytotoxicity was assayed after incubation of the islet cells with fresh human serum. Furthermore, adult islets from normal control pigs and hDAF TG pigs were exposed to fresh human blood in vitro for 60 min, and the inflammatory reaction elicited was compared between the different types of islets. RESULTS: All human islet cell preparations expressed hCD59, two of five expressed hMCP, but none expressed hDAF. Porcine islet cells expressed both pCD59 and pMCP. Normal adult porcine islet cells exposed to fresh human serum resulted in 74+/ 5.4% cell lysis (mean+/-SEM, n=16). In comparison, only 1.3+/-2.8% (n=20, P<0.001) of human islet cells were lysed in the human serum. One islet cell preparation from an hDAF TG pig expressed small amounts of hDAF. This preparation from hDAF TG pigs bound significantly less C3c than did normal control islets (mean fluorescence ratio 16+/-2.2 and 58+/-4.3, respectively; P=0.046) and were partially protected from cell lysis in fresh human serum (47+/-10% and 78+/-18% cell lysis, respectively; P=0.046). The other four preparations from hDAF TG pigs were negative for hDAF and were equally susceptible to lysis as normal control islets. All fetal pancreatic islet cells from hDAF TG pigs analyzed were negative for hDAF expression. When exposed to fresh human blood in vitro, adult and fetal islets from hDAF TG pigs elicited equally strong inflammatory changes as did the normal control islets. The inflammatory changes were characterized by activation of the complement and coagulation systems, resulting in islet damage with "dumping" of insulin into the blood. CONCLUSIONS: Porcine and human islet cells express species-restricted complement regulatory proteins, with the human islet cells expressing mainly hCD59. A low expression of hDAF was detected on islet cells from one of five hDAF TG pigs. These islet cells displayed reduced islet cell cytotoxicity in fresh human serum. We conclude that protection from complement-mediated lysis will be important in the context of intraportal pig-to human islet transplantation, and expression of a human RCA on islet cells should be beneficial in this context. PMID- 11477360 TI - A comparison of gene expression in murine cardiac allografts and isografts by means DNA microarray analysis. AB - BACKGROUND: Acute rejection of allografts remains a significant problem in clinical transplantation, and the fundamental mechanism underlying this rejection are as yet only poorly elucidated. Recently, DNA microarrays have come into use for the study of gene expression profiles, and we have taken advantage of this new technology to investigate acute rejection. We compared mRNA profiles in murine cardiac allografts with isografts using DNA microarrays with probe sets corresponding to more than 11,000 mice genes. METHODS: We screened for gene expression changes in murine cardiac allografts between fully incompatible mice strains (BALB/c H2d to C3H/He H2k) using a DNA microarray. The heart was heterotopically transplanted. Allografts (BALB/c to C3H/He) were removed on days 1, 3, and 5. As a control, isografts (C3H/He to C3H/He) harvested on days 1, 3, and 5 and native hearts of both strain mice (C3H/He and BALB/c) were obtained. RESULTS: On day 5, interferon-gamma (IFN-gamma) and many IFN-gamma-inducible genes were profoundly induced in the allograft relative to isograft. Monokine induced by IFN-gamma was most profoundly induced followed by inducibly expressed GTPase and Lmp-2. IFN-gamma was also profoundly induced. The induction was detectable from day 3. In contrast, genes regulated by other cytokines exhibited only modest changes. CONCLUSION: IFN-gamma-inducible genes are specifically up regulated in murine cardiac allografts, suggesting that signaling mediated by IFN gamma may play an important role in the late phase of acute rejection in vivo. PMID- 11477361 TI - Antimurine immunoglobulin antibody responses after the administration of murine monoclonal antibodies to rats are altered by small bowel allograft rejection. AB - BACKGROUND: This study monitored the induction of antimurine immunoglobulin antibody responses after the administration of anti-CD4 (OX38) and anti-LFA-1 (WT.1) monoclonal antibodies to DA rats. METHODS: Monoclonal antibody was administered i.v. on 3 consecutive days to untransplanted DA rats, and DA recipients of PVG small bowel allografts. Control animals received no monoclonal antibody. Antimurine immunoglobulin antibody levels in serum samples were determined by enzyme immunoassay. RESULTS: No antimurine immunoglobulin antibody was detected in untransplanted animals receiving OX38 alone. Reactivity was apparent in WT.1-treated animals, but this response was totally abrogated by the co-administration of OX38. A combination of OX38 and WT.1 had no effect on allograft recipient survival and antimurine immunoglobulin antibody responses were detected in all allograft recipients, irrespective of the treatment regimen. CONCLUSIONS: Although OX38 inhibited the antibody response both to itself and to WT.1 in untransplanted animals, the immune reaction induced by small bowel allograft rejection overcame this inhibitory capacity. PMID- 11477362 TI - Efficacy of lamivudine for the treatment of hepatitis B virus infection after liver transplantation in children. AB - BACKGROUND: There is at present very little information about hepatitis B virus (HBV) infection in children after liver transplantation. This is the first study to assess the safety and efficacy of lamivudine in this patient population. METHODS: We describe three children aged 5-14 years who underwent liver transplantation for fulminant hepatitis A, hyperoxaluria, and cystic fibrosis. Despite adequate immunoprophylaxis, two of the children who were serum hepatitis B surface antigen-positive before transplantation (HBV DNA-negative by hybridization) had a reactivation of the disease, and one had a de novo HBV infection, at 12-18 months after transplantation. Lamivudine 3 mg/kg was administered on a compassionate-use basis for 14-36 months. RESULTS: After 1 month of therapy, HBV DNA disappeared from the serum in all patients by hybridization and in two patients by polymerase chain reaction. In all three children, alanine transaminase levels normalized. One child developed lamivudine resistance after 22 months with no evidence of hepatic decompensation. Repeated liver histological studies revealed progression of hepatic fibrosis in one child. All children remained serum hepatitis B surface antigen- and hepatitis B e antigen-positive. No adverse effects of the drug were noted. CONCLUSION: Lamivudine is beneficial and well tolerated in children with HBV infection after liver transplantation. PMID- 11477363 TI - Pseudoaneurysm of the superior mesenteric artery after pancreas transplantation treated by endovascular stenting. AB - Pseudoaneurysm after pancreas transplantation can have serious consequences, including rupture, hemorrhage, and graft loss. We describe a 38-year-old patient who presented with a pseudoaneurysm of the donor superior mesenteric artery 1 month after pancreas transplantation. Selective arteriography was performed and the lesion was repaired with endovascular placement of a 28-mm covered stent. Laparotomy was avoided. The pancreatic graft was continuing to function well 9 months later. As far as we know, this minimally invasive approach was not previously reported. According to published series, pseudoaneurysms often occur secondary to infection and require operative intervention necessitating graft pancreatectomy. Patients can present with serious symptoms including hypotension and shock. Therefore, it is important to detect pseudoaneurysm in a timely manner. Computed tomography and Doppler ultrasound are important diagnostic tools in this regard. We demonstrated the utility of endovascular stenting in the treatment of pseudoaneurysm after pancreas transplantation. When used in a timely manner in well selected patients, endovascular stenting can abrogate the need for operative intervention and its attendant morbidity. PMID- 11477364 TI - Bile leak from duct of Luschka after liver transplantation. AB - BACKGROUND: We report a case of bile leak from an accessory duct of Luschka during cholecystectomy during liver transplantation. METHODS: Radiological findings suggested that the collection was septated. An intra-operative cholangiogram was obtained by cannulation of the accessory hepatic duct. RESULTS: An infected biloma with Clostridium perfringens was drained surgically. The bile leak that emanated from the gall bladder fossa was found to communicate with an accessory right hepatic duct draining a segmental duct in the right liver lobe. The bile leak resolved completely after direct suture of the accessory duct. CONCLUSIONS: Excessive use of electrocautery to the liver bed during donor cholecystectomy may injure subcapsular ducts in the gallbladder fossa. In liver transplantation, dissection should be kept close to the serosal lining of the gall bladder, preserving the areolar tissue in the gall bladder bed, to avoid injury to the duct of Luschka. PMID- 11477365 TI - Will mixed chimerism cure autoimmune diseases after a nonmyeloablative stem cell transplant? AB - BACKGROUND: Mixed chimerism after allogeneic bone marrow transplantation has been shown to cure a number of genetic disorders in both the clinical and experimental settings. Although encouraging results have been reported from animal experiments, the role of mixed chimerism in eliminating autoimmune disorders is not clear. METHODS: A 50-year-old man with extensive psoriasis received an allogeneic transplant from his brother after nonmyeloablative conditioning with fludarabine, melphalan, and Campath-1H for relapsed non-Hodgkin's lymphoma. The chimerism status and the immunological recovery after the transplant were serially monitored. RESULTS: Twenty-one months after the transplant, the patient continues to be in complete remission from psoriasis and lymphoma with stable mixed chimerism (30% to 40% donor cells), despite significant recovery of T-cell subsets and antigen-specific response. CONCLUSIONS: If mixed chimerism can be achieved safely with novel low-intensity conditioning regimens and results in sustained remission of autoimmune diseases, allogeneic transplantation may become a realistic therapy in the management of some patients with autoimmune disease. PMID- 11477366 TI - Multiple adenomas and hepatocellular carcinoma in a renal transplant patient with glycogen storage disease type 1a (von Gierke disease). AB - We report on a 42-year-old female patient with glycogen storage disease type 1a (von Gierke disease, GSD 1a) who developed hepatic adenomas and finally a hepatocellular carcinoma 10 years after renal transplantation. The tumor was resected; however, the patient died 6 months later as a result of fulminant carcinoma recurrence. In patients who have GSD 1a with terminal renal failure, combined liver and kidney transplantation may be considered at an early stage of the disease. PMID- 11477367 TI - Elevated right ventricular pressures are not a contraindication to liver transplantation in Alagille syndrome. AB - BACKGROUND: Elevated right ventricle pressure resulting from pulmonary artery stenoses may affect outcome and survival after liver transplantation in patients with Alagille syndrome. METHODS AND RESULTS: Between 1984 and 1997, among 444 pediatric liver transplant recipients, 17 had liver transplantation for Alagille syndrome (mean age 3.5 years, range 1.2-13 years), mainly because of poor quality of life with intractable pruritus, and failure to thrive. All patients had pulmonary artery stenosis. In 10 patients considered to have elevated RV pressure on ECG and/or Doppler-echocardiography, a cardiac catheterization was performed before liver transplantation. Mean RV systolic pressure was 55 mmHg (median 49.5 mmHg, range 35-98 mm Hg), mean RV to left ventricular systolic pressure ratio 0.53 (median 0.53, range 0.29-0.78) with a ratio above 0.5 in 6 patients (median 0.66, range 0.5-0.8). All patients underwent successful liver transplantation. Five patients died 1 to 9 months after transplantation from noncardiac causes. In two of them, cardiac catheterization before transplantation showed a RV to left ventricular pressure ratio of 0.51 in one and 0.37 in the second. In the three others, echocardiography before transplantation estimated RV pressures below 0.5 systemic pressures. At follow-up (median 6 years, range 1.5-15 years), liver tests were normal in all, none complained of pruritus and body weight was normalized in 70%. None of the patients presented cardiac symptoms, arrhythmias, or worsening of their cardiac status. CONCLUSIONS: Liver transplantation can be performed safely in children with Alagille syndrome, even in the presence of elevated right ventricular pressure. PMID- 11477368 TI - Interleukin-17 antagonism inhibits acute but not chronic vascular rejection. AB - BACKGROUND: Blocking the action of interleukin (IL) 17 with an IL-17 receptor (R):Fc fusion protein inhibits T-cell proliferative responses to alloantigens and prolongs vascularized heart graft survival. In this study, we examined whether IL 17 antagonism could suppress the development of chronic rejection. METHODS: A 0.6 cm section of C57BL10 (H2b) thoracic aorta was transplanted to recipient C3H (H2k) abdominal aorta. IL-17R:Fc or control human immunoglobulin G was administered i.p. (500 microg/day) from days 0 to 6 or from days 0 to 29. Mice were killed on days 7 or 30. Grafts were examined histologically and stained for alpha-smooth muscle actin (alpha-smA). Antidonor mixed leukocyte reaction, cytotoxic T cell, and alloantibody responses were quantified. RESULTS: On day 7, control grafts showed mononuclear cell (MNC) infiltration, pronounced endothelial damage, and apoptosis of intimal and medial cell compartments. By day 30, there was concentric intimal thickening, accumulation of alpha-smA+ cells, and collagen deposition. Patchy destruction of the elastic membranes and loss of alpha-smA expression in media were evident. IL-17R:Fc for 6 days decreased MNC infiltration in the intimal and medial compartments at day 7. The endothelium was preserved (completely or partially) in all grafts. The medial compartment showed normal alpha-smA expression. Irrespective of IL-17R:Fc treatment for either 6 days or continuously, allografts harvested at day 30 showed circumferential intimal thickening, with accumulation of alpha-smA+ cells and collagen deposition. There was no effect on circulating alloantibody levels. CONCLUSIONS: These findings support a role for IL-17 in the immunopathogenesis of acute vascular rejection and demonstrate the potential of IL-17 antagonism for therapy. By contrast, IL-17 antagonism does not appear to prevent ensuing chronic graft vascular disease, in particular neointimal formation. PMID- 11477369 TI - Long-term islet allograft function in the absence of chronic immunosuppression: a case report of a nonhuman primate previously made tolerant to a renal allograft from the same donor. AB - Development of mixed chimerism by donor bone marrow transplantation (DBMT) has led to long-term tolerance of solid organ allografts in nonhuman primates. As an initial attempt to extend this approach to cellular transplant, islet transplant from the same donor was attempted in the recipient previously made tolerant to a kidney allograft. METHODS: After the conditioning with ATG, total body irradiation, thymic irradiation, and splenectomy, DBMT was performed followed by 4 weeks of cyclosporine. Kidney transplantation and native nephrectomies were subsequently performed on day 89. After 2.8 years of DBMT, diabetes was induced by streptozocin (STZ) and islets from bone marrow and kidney donor were transplanted without immunosuppression. RESULTS: After DBMT, the recipient developed chimerism and no evidence of kidney rejection for more than 1000 days. STZ induced diabetes was reversed after the islet transplantation. Islet biopsies demonstrated insulin staining without rejection. Although the recipient became diabetic 300 days after islet transplantation, viable transplanted islets were found in the liver and under the kidney capsule without any evidence of rejection. CONCLUSION: Tolerance with a nonmyeloablative conditioning can allow successful pancreatic islet transplantation without immunosuppression. Because no histological evidence of rejection was identified, recurrent diabetes is presumed to be inadequate islet mass. PMID- 11477370 TI - Topsy turvydom in brain death determination. PMID- 11477371 TI - Recipient outcome after receiving a laparoscopically harvested kidney from a living donor. PMID- 11477373 TI - Cancer of the oral cavity. PMID- 11477374 TI - Metabolism of rabbit angiostatin glycoforms I and II in rabbits: angiostatin-I leaves the intravascular space faster and appears to have greater anti-angiogenic activity than angiostatin-II. AB - Plasminogen (PLG) exists in the circulation as two glycoforms, I and II. Angiostatin (AST) is a polypeptide that has been cleaved from the kringle region of PLG and has strong anti-angiogenic properties. AST-I and AST-II, which consisted only of kringles 1 through 3, were prepared by the action of urokinase on purified rabbit PLG-I and PLG-II, respectively, in the presence of N-acetyl cysteine, followed by affinity chromatography on lysine-Sepharose. Purified AST-I and AST-II were tested for functional activity with a chick chorioallantoic membrane (CAM) model; when similar amounts were applied to a 6-day CAM, AST-I was substantially more effective than AST-II in decreasing vascular supply to the CAM over a 72-hour period; this activity correlated with a loss of capillaries, probably through apoptosis of endothelial cells. Radiolabeled AST-I and AST-II (iodine 125 and iodine 131) were co-injected intravenously into healthy rabbits to determine their clearances from plasma measured over 3 days. Over a dose range of 0.08 to 2.7 microg/kg, the fractional catabolic rate within the intravascular space (j(3)) indicated that AST-I was cleared 3-fold to 4-fold more rapidly than AST-II (P < .001). The catabolic half-life of AST-I (2.01 +/- 0.19 days) was significantly less than that of AST-II (2.62 +/- 0.20 days). The faster clearance of AST-I from the intravascular space was matched by its more rapid passage than AST-II to the extravascular space of various organs over 60 minutes in vivo. This property of AST-I as compared with AST-II may partially explain its greater anti angiogenic potential. From the plasma concentrations of PLG-I and PLG-II and their relative behaviors toward rabbit VX-2 lung tumors in vivo, we predict that substantially greater quantities of AST-II than AST-I may be released into the extravascular space of tumors. PMID- 11477375 TI - Acute phase response and energy balance in stable human immunodeficiency virus infected patients: a doubly labeled water study. AB - We sought to assess possible associations of the acute phase response with energy balance as a cause of malnutrition in uncomplicated HIV-infection. Seven stable HIV-infected patients and 7 control subjects were followed for 2 weeks for blood chemistry, body composition, total and resting energy expenditure (TEE, REE), heart rate, energy intake, and physical activity; 6 patients were investigated for interleukin-2 receptor. TEE, REE, energy intake, and anthropometric data in patients and control subjects were very similar. However, physical activity, total body potassium, and bioimpedance phase angle were decreased (1.41 +/- 0.08 vs 1.55 +/- 0.9, 152 +/- 10 g vs 191 +/- 37 g, and 6.4 +/- 0.8 degrees vs 7.1 +/- 0.5 degrees; each P < .05), and mean heart rate, fibrinogen, and erythrocyte sedimentation rate were increased in HIV-infected patients (84 +/- 6 bpm vs 76 +/ 8 bpm, 4.3 +/- 1.2 g/L vs 2.5 +/- 0.4 g/L, and 21 +/- 13 mm vs 2 +/- 3 mm; P < .05). The deviation between the measured and the predicted REE in the patient group correlated positively with heart rate and serum interleukin-2 receptor concentrations (r = 0.83 and r = 0.91; P < .05). Possible increases in REE caused by an ongoing acute phase response may be counterbalanced by reduced physical activity that results in normal TEE in HIV infection. PMID- 11477376 TI - Inhibition of mesothelioma cell growth in vitro by doxycycline. AB - Malignant mesothelioma causes profound morbidity and nearly universal mortality that is often refractory to conventional treatment modalities of aggressive surgery, radiotherapy, or chemotherapy. Doxycycline, a commonly used antibiotic, has anti-tumor activity against several malignancies, but its anti-tumor effects on malignant mesothelioma have not been evaluated. We report here that concentrations of doxycycline achievable in serum with typical oral doses had cytostatic effects to varying extent on all eight of the mesothelioma cell lines studied but did not affect normal lung fibroblasts. Doxycycline inhibited the production of mitochondrial cytochrome c oxidase, especially in mesothelioma cells more sensitive to its cytostatic effects, and directly inhibited gelatinase A activity; both of these activities are putative mechanisms for the cytostatic activity of doxycycline in other tumor cells. Thus doxycycline may have a role as adjuvant therapy for malignant mesothelioma. PMID- 11477377 TI - Inhibition of the second phase of amyloidogenesis in a mouse model by a single dose colchicine regimen. AB - Amyloidogenesis consists of two stages. In the first, amyloid enhancing factor (AEF) is generated, and in the second, deposition of amyloid fibrils occurs. Colchicine is a known inhibitor of amyloidosis of familial Mediterranean fever (FMF) and of mouse experimental amyloidosis, but the timing and mechanism of its effect are still unclear. The aim of this study is to determine whether colchicine inhibits the second phase of amyloidogenesis and to study the time correlate of such an effect. To that end, amyloid was induced in Swiss male mice with AEF and AgNO(3) (an inflammatory stimulus), a method that skips the first phase of amyloidogenesis. Two amyloid induction protocols were used: a standard protocol, in which AEF and AgNO(3) were administered concurrently, and a prolonged protocol, in which the administration of AgNO(3) was delayed by 24 hours or 7 days. To study the inhibitory effect of colchicine on the second phase of amyloidogenesis, a single dose of colchicine (30 microg) was injected intravenously before, during, or after administration of AgNO(3) in both the standard and prolonged amyloid induction protocols. The amount of amyloid deposition in the spleens was determined with the crush-and-smear technique and a 5-grade scale. Colchicine was found to inhibit the second phase of amyloidogenesis. Its best effect was achieved when administered 48 hours after initiation of AgNO(3) injections. The pattern of colchicine-inhibition-in-time in the standard and the prolonged amyloid induction protocols was similar, indicating that colchicine exerts inhibition through its effect on the inflammatory stimulus (AgNO(3)). These findings suggest that (1) colchicine suppresses amyloidogenesis in the late (second) stage and that (2) this suppression is possibly related to the anti-inflammatory effect of colchicine. PMID- 11477378 TI - Interplay of antibody and T cell responses in acute myocardial infarction. AB - This study sought to investigate the interplay between antibody and T cell responses triggered by an acute myocardial infarction (MI) and their possible role in the progress of this disease. Serum samples were collected from two groups of patients, group A (n = 26) within the first week of MI, and group B (n = 28) at 2 weeks and 2 months after MI. Patients in group A were older and had higher prevalence of hypertension and previous attack of MI than patients in group B. The levels of anti-myosin immunoglobulin M and immunoglobulin G antibodies in the serum samples from group A were significantly higher than those in normal control subjects. In group B, the levels of both antibodies were lower than those in group A but remained significantly higher than those in normal control subjects at both 2 weeks and 2 months. The levels of intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in the serum samples from group A patients were significantly higher than those in normal control subjects. At 2 weeks after MI (group B), only the level of sVCAM 1, but not that of sICAM-1, was significantly higher than that in normal control subjects, and there were no significant changes in the levels of these two molecules from 2 weeks to 2 months after MI. We conclude that the higher levels of anti-myosin antibodies and adhesion molecules in group A patients as compared with group B patients may be due to higher or more frequent exposures of their immune systems to heart antigens. Furthermore, the immunoglobulin M antibody response during the first week of MI had an inverse relationship with the level of interleukin-2R (sIL-2R), which suggested a possible suppressive or regulatory role of this antibody on the cellular immune response during this time. PMID- 11477379 TI - Role of neuropeptide Y in renal sympathetic vasoconstriction: studies in normal and congestive heart failure rats. AB - Sympathetic nerve activity, including that in the kidney, is increased in heart failure with increased plasma concentrations of norepinephrine and the vasoconstrictor cotransmitter neuropeptide Y (NPY). We examined the contribution of NPY to sympathetically mediated alterations in kidney function in normal and heart failure rats. Heart failure rats were created by left coronary ligation and myocardial infarction. In anesthetized normal rats, the NPY Y(1) receptor antagonist, H 409/22, at two doses, had no effect on heart rate, arterial pressure, or renal hemodynamic and excretory function. In conscious severe heart failure rats, high-dose H 409/22 decreased mean arterial pressure by 8 +/- 2 mm Hg but had no effect in normal and mild heart failure rats. During graded frequency renal sympathetic nerve stimulation (0 to 10 Hz), high-dose H 409/22 attenuated the decreases in renal blood flow only at 10 Hz (-36% +/- 5%, P <.05) in normal rats but did so at both 4 (-29% +/- 4%, P <.05) and 10 Hz (-33% +/- 5%, P <.05) in heart failure rats. The glomerular filtration rate, urinary flow rate, and sodium excretion responses to renal sympathetic nerve stimulation were not affected by high-dose H 409/22 in either normal or heart failure rats. NPY does not participate in the regulation of kidney function and arterial pressure in normal conscious or anesthetized rats. When sympathetic nervous system activity is increased, as in heart failure and intense renal sympathetic nerve stimulation, respectively, a small contribution of NPY to maintenance of arterial pressure and to sympathetic renal vasoconstrictor responses may be identified. PMID- 11477380 TI - Exploring the "iron shuttle" hypothesis in chelation therapy: effects of combined deferoxamine and deferiprone treatment in hypertransfused rats with labeled iron stores and in iron-loaded rat heart cells in culture. AB - Although iron chelation therapy results in a significant improvement in well being and life expectancy of thalassemic patients with transfusional iron overload, failure to achieve these goals in a substantial proportion of patients underlines the need for improved methods of treatment. In the present studies we used selective radioactive iron probes of hepatocellular and reticuloendothelial (RE) iron stores in hypertransfused rats and iron-loaded heart cells to compare the source of iron chelated in vivo by deferoxamine (DFO) or by deferiprone (L1) and its mode of excretion, to examine the ability of DFO and L1 to remove iron directly from iron-loaded myocardial cells, and to examine the mechanism of their combined interaction through a possible additive or synergistic effect. Our results indicate that L1 given orally is 1.6 to 1.9 times more effective in rats, on a weight-per-weight basis, than parenteral DFO in promoting the excretion of storage iron from parenchymal iron stores but shows no advantage over DFO in promoting RE iron excretion. Simultaneous administration of DFO and L1 results in an increase in chelating effect that is additive but not synergistic. The magnitude of this additive effect is identical to an increase in the equivalent (weight or molar) dose of DFO alone rather than the sum of the separate effects of L1 and DFO. This finding is most probably the result of a transfer of chelated iron from L1 to DFO. These observations may have practical implications for current efforts to design better therapeutic strategies for the management of transfusional iron overload. PMID- 11477381 TI - Contact between a polymer and whole blood: sequence of events leading to thrombin generation. AB - The mechanism by which thrombin is generated on a polymer surface in an extracorporeal circuit is not yet fully understood. To address this question we have developed an in vitro chamber model in which whole blood containing heparin (1 IU/mL) comes in contact with a commonly used biomaterial, polyvinyl chloride (PVC). Incubation of blood in the chamber for 60 minutes at 37 degrees C resulted in the binding of platelets to the material surface and the generation of thrombin-antithrombin complexes. Corn trypsin inhibitor, a specific inhibitor of factor XIIa, inhibited this thrombin-antithrombin complex generation in blood in contact with PVC, which is not considered an efficient activator of factor XII. The addition of the glycoprotein IIb/IIIa inhibitor Ro44-9883 abrogated platelet binding and aggregation and resulted in decreased generation of thrombin antithrombin complexes. Thrombin-antithrombin generation was also negligible in platelet-rich plasma but could be partially restored in the presence of erythrocytes. Taken together, these data are compatible with a model in which thrombin generation is triggered by factor XII. The response to contact with PVC appears to begin with a low-grade generation of thrombin that involves both erythrocytes and leukocytes and that activates platelets, followed by the activation of a platelet-dependent amplification loop that produces most of the thrombin. PMID- 11477382 TI - Hematocrit as a function of age and altitude in China. PMID- 11477385 TI - A sporting difference. PMID- 11477386 TI - Rural trauma nursing: life in the vast lane... PMID- 11477387 TI - Case report: survival of a type I transthoracic impalement. AB - The care of a patient who became impaled on a large aluminum pipe is presented. A review of the literature reveals that most patients with a type I injury either do not survive or present with an unpredictable pattern of injury. Preoperative care requires rapid stabilization, assessment, and interventions based on the pattern of injury. Perioperative management may involve multiple surgeons performing simultaneous surgical procedures. PMID- 11477388 TI - Common factors of spontaneous self-extubation in a critical care setting. AB - A prospective, concurrent study was conducted of all patients who self-extubated in a mixed critical care setting during a 14-month period. The purpose of the study was to identify the incidence and common factors associated with spontaneous self-extubation (SSE). A total of 75 cases of SSE occurred in 68 patients who had an incidence of 38.5 SSEs per 100 intubated days. The analysis of common factors of the total population found the following: 60 cases (80%) were restrained; 44 cases (59%) required reintubation; 66 cases (88%) followed commands or localized painful stimuli at the time of SSE; and 67 cases (89%) elicited spontaneous eye opening or opened eyes to verbal command at the time of SSE. Only 18 cases (24%) had analgesia administered within 1 to 2 hours of SSE. Twenty-four cases (32%) had anxiolytics administered within 4 hours of SSE. Of the 56 cases of SSE that were witnessed, 43 cases (73% of those observed) were considered deliberate rather than accidental. The practice of using intravenous boluses on an "as needed" dosing frequency for administering sedation and analgesia was a common factor in SSE. Adequate doses of sedation and analgesia delivered by continuous infusion may prevent SSE in alert, intubated patients. PMID- 11477389 TI - Safety City: babysitter guide. PMID- 11477390 TI - Nursing protocol: artificial airway management. PMID- 11477391 TI - Wheeled recreation resources: bikes, in-line skates, scooters, and skateboards. PMID- 11477392 TI - Cardiac perforation and tamponade: the deadly duo of central venous catheters. PMID- 11477394 TI - Accidentally informed. PMID- 11477393 TI - In-line skates and skateboards. PMID- 11477395 TI - New nomenclature for MHC receptors. PMID- 11477396 TI - Ruby anniversary: forty years of thymus immunology research. PMID- 11477397 TI - SAP: natural inhibitor or grand SLAM of T cell activation? PMID- 11477398 TI - Lung Krupple-like factor: a quintessential player in T cell quiescence. PMID- 11477399 TI - TCR avidity: it's not how strong you make it, it's how you make it strong. PMID- 11477400 TI - The mucosal milieu creates tolerogenic dendritic cells and T(R)1 and T(H)3 regulatory cells. PMID- 11477402 TI - Toll-like receptors: critical proteins linking innate and acquired immunity. AB - Recognition of pathogens is mediated by a set of germline-encoded receptors that are referred to as pattern-recognition receptors (PRRs). These receptors recognize conserved molecular patterns (pathogen-associated molecular patterns), which are shared by large groups of microorganisms. Toll-like receptors (TLRs) function as the PRRs in mammals and play an essential role in the recognition of microbial components. The TLRs may also recognize endogenous ligands induced during the inflammatory response. Similar cytoplasmic domains allow TLRs to use the same signaling molecules used by the interleukin 1 receptors (IL-1Rs): these include MyD88, IL-1R--associated protein kinase and tumor necrosis factor receptor--activated factor 6. However, evidence is accumulating that the signaling pathways associated with each TLR are not identical and may, therefore, result in different biological responses. PMID- 11477403 TI - Regulation of SLAM-mediated signal transduction by SAP, the X-linked lymphoproliferative gene product. AB - Signaling lymphocyte activation molecule (SLAM)-associated protein (SAP) is a short intracellular molecule that is mutated in humans with X-linked lymphoproliferative (XLP) disease. Although the exact role and mechanism of action of SAP are not known, it has the capacity to interact with the cytoplasmic region of SLAM and other related immune cell receptors. As SAP is composed almost exclusively of a Src homology 2 (SH2) domain, it has been proposed that it functions as a natural blocker of SH2 domain--mediated interactions. We report here that the SLAM receptor is capable of triggering a protein tyrosine phosphorylation signal in T cells via a mechanism that is strictly dependent on SAP expression. This signal involves the SH2 domain--containing inositol phosphatase (SHIP); the adaptor molecules Dok2, Dok1 and Shc; and Ras GTPase- activating protein RasGAP. SAP is essential for this pathway because it facilitates the selective recruitment and activation of the Src-related protein tyrosine kinase FynT. We also show that signaling via the SLAM-SAP pathway in an established T cell line can alter the profile of cytokine production during T cell activation. These findings identify a mechanism by which a putative adaptor molecule is required for receptor-mediated signaling events in the immune system. They also provide insights into the pathophysiology of a severe human lymphoproliferative disease. PMID- 11477404 TI - The beta-catenin--TCF-1 pathway ensures CD4(+)CD8(+) thymocyte survival. AB - The association of trans-acting T cell factors (TCFs) or lymphoid enhancer factor 1 (LEF-1) with their coactivator beta-catenin mediates transient transcriptional responses to extracellular Wnt signals. We show here that T cell maturation depends on the presence of the beta-catenin--binding domain in TCF-1. This domain is necessary to mediate the survival of immature CD4(+)CD8(+) double-positive (DP) thymocytes. Accelerated spontaneous thymocyte death in the absence of TCF-1 correlates with aberrantly low expression of the anti-apoptotic protein Bcl-x(L). Increasing anti-apoptotic effectors in thymocytes by the use of a Bcl-2 transgene rescued TCF-1-deficient DP thymocytes from apoptosis. Thus, TCF-1, upon association with beta-catenin, transiently ensures the survival of immature T cells, which enables them to generate and edit T cell receptor (TCR) alpha chains and attempt TCR-mediated positive selection. PMID- 11477405 TI - Transcription factor LKLF is sufficient to program T cell quiescence via a c-Myc dependent pathway. AB - T lymphocytes circulate in a quiescent state until they encounter cognate antigen bound to the surface of an antigen-presenting cell. The molecular pathways that regulate T cell quiescence remain largely unknown. Here we show that forced expression of the lung Kruppel-like transcription factor (LKLF) in Jurkat T cells is sufficient to program a quiescent phenotype characterized by decreased proliferation, reduced cell size and protein synthesis and decreased surface expression of activation markers. Conversely, LKLF-deficient peripheral T cells produced by gene targeting showed increased proliferation, increased cell size and enhanced expression of surface activation markers in vivo. LKLF appeared to function, at least in part, by decreasing expression of the proto-oncogene encoding c-Myc. Forced expression of LKLF was associated with markedly decreased c-Myc expression. In addition, many effects of LKLF expression were mimicked by expression of the dominant-negative MadMyc protein and rescued by overexpression of c-Myc. Thus, LKLF is both necessary and sufficient to program quiescence in T cells and functions, in part, by negatively regulating a c-Myc--dependent pathway. PMID- 11477406 TI - CD4(+) T cell effectors can become memory cells with high efficiency and without further division. AB - Whether memory T lymphocytes are derived directly from effector T cells or via a separately controlled pathway has long been debated. Here we present evidence that, after adoptive transfer, a large fraction of in vitro--derived effector CD4(+) T cells have the potential to become memory T cells and that this transition can occur without further division. This data supports a linear pathway from effector to memory cells and suggests that most properties of memory cells are predetermined during effector generation. We suggest, therefore, that evaluation of vaccine efficacy in the induction of memory CD4(+) T cells should focus on the effector stage. PMID- 11477407 TI - Functional avidity maturation of CD8(+) T cells without selection of higher affinity TCR. AB - Unlike B cells, T cells lack the capacity to improve the affinity of their antigen receptors by somatic mutation. It is, therefore, believed that optimization of cellular immunity is mediated almost exclusively through selective expansion of T cells bearing receptors with the highest affinity for antigen. We show here that T cell responsiveness to peptide (termed "functional avidity") increased>50-fold during the early stages of viral infection. This indicated that T cells, like B cells, undergo extensive functional maturation in vivo. However, in contrast to B cells, maturation of the T cell response can occur without any appreciable change in T cell receptor affinity. PMID- 11477408 TI - Identification of very early lymphoid precursors in bone marrow and their regulation by estrogen. AB - Estrogen is a negative regulator of lymphopoiesis and provides an experimental tool for probing relationships between lymphocyte precursors and stem cells. We found that expression of lymphocyte-associated genes and immunoglobulin (Ig) gene rearrangement occurred before CD45R acquisition. Lymphoid-restricted progenitors that were Lin(-)IL-7R alpha(+)c-kit(lo)TdT(+) (lineage marker(-), interleukin receptor 7 alpha(+), c-kit(lo) and terminal deoxynucleotidyl transferase(+)) were selectively depleted in estrogen-treated mice; within a less differentiated Lin-c kit(hi) fraction, functional precursors of B and T, but not myeloid, cells were also selectively depleted. TdT and an Ig heavy chain transgene were detected within a hormone-regulated Lin(-)c-kit(hi)Sca-1(+)CD27(+)Flk-2(+)IL-7R alpha(-) subset of this multipotential progenitor population. Identification of these extremely early lymphoid precursors should facilitate investigation of the molecular mechanisms that control lineage-fate decisions in hematopoiesis. PMID- 11477409 TI - Pulmonary dendritic cells producing IL-10 mediate tolerance induced by respiratory exposure to antigen. AB - Respiratory exposure to allergen induces T cell tolerance and protection against the development of airway hyperreactivity and asthma. However, the specific mechanisms by which tolerance is induced by respiratory allergen are not clear. We report here that pulmonary dendritic cells (DCs) from mice exposed to respiratory antigen transiently produced interleukin 10 (IL-10). These phenotypically mature pulmonary DCs, which were B-7(hi) as well as producing IL 10, stimulated the development of CD4(+) T regulatory 1--like cells that also produced high amounts of IL-10. In addition, adoptive transfer of pulmonary DCs from IL-10(+/+), but not IL-10(-/-), mice exposed to respiratory antigen induced antigen-specific unresponsiveness in recipient mice. These studies show that IL 10 production by DCs is critical for the induction of tolerance, and that phenotypically mature pulmonary DCs mediate tolerance induced by respiratory exposure to antigen. PMID- 11477410 TI - CX3C chemokine mimicry by respiratory syncytial virus G glycoprotein. AB - Chemokines are chemoattractant proteins that are divided into subfamilies based upon cysteine signature motifs termed C, CC, CXC and CX3C. Chemokines have roles in immunity and inflammation that affect cell trafficking and activation of T cells as well as cells of the innate immune system. We report here CX3C chemokine mimicry for the G glycoprotein of respiratory syncytial virus (RSV) and show binding to CX3CR1--the specific receptor for the CX3C chemokine fractalkine--and induction of leukocyte chemotaxis. We also show that CX3CR1 facilitates RSV infection of cells. Thus, G glycoprotein interaction with CX3CR1 probably plays a key role in the biology of RSV infection. PMID- 11477411 TI - Contribution of the innate immune system to autoimmune myocarditis: a role for complement. AB - Myocarditis is a principal cause of heart disease among young adults and is often a precursor of heart failure due to dilated cardiomyopathy. We show here that complement is critical for the induction of experimental autoimmune myocarditis and that it acts through complement receptor type 1 (CR1) and type 2 (CR2). We also found a subset of CD44(hi)CD62L(lo) T cells that expresses CR1 and CR2 and propose that both receptors are involved in the expression of B and T cell activation markers, T cell proliferation and cytokine production. These findings provide a mechanism by which activated complement, a key product of the innate immune response, modulates the induction of an autoimmune disease. PMID- 11477412 TI - Autoantibodies to GPI in rheumatoid arthritis: linkage between an animal model and human disease. AB - In K/BxN T cell receptor-transgenic mice, spontaneous inflammatory arthritis exhibiting many of the features of human rheumatoid arthritis (RA) is initiated by T cells, but is almost entirely sustained by antibodies to the self-antigen glucose-6-phosphate isomerase (GPI). The relevance of these observations to human disease has been questioned. Here we show that 64% of humans with RA, but not controls, had increased concentrations of anti-GPI immunoglobulin G (IgG) in serum and synovial fluid. In addition, the concentrations of soluble GPI in the sera and synovial fluids of RA patients were also elevated, which led to immune complex formation. Using phage-display methods, we cloned a panel of specific high-affinity human monoclonal anti-GPI IgGs from a patient with RA. These antibodies were highly somatically mutated, which was indicative of an affinity matured response that was antigen driven. Immunohistochemistry of RA synovium showed high concentrations of GPI on the surface of the synovial lining and on the endothelial cell surface of arterioles; this indicated a mechanism by which antibodies to GPI may precipitate joint disease. The results indicate that the immunological events that lead to the development of autoimmune disease in the K/BxN mouse model may also occur in human RA. This data may be used to develop new strategies for therapeutic intervention. PMID- 11477413 TI - Online in less time. PMID- 11477414 TI - Our 2000 impact factor. PMID- 11477415 TI - Molecular stepping stones in memory consolidation. PMID- 11477416 TI - Ion-age molecular motors. PMID- 11477417 TI - Exciting glial oscillations. PMID- 11477418 TI - Faces and races in the brain. PMID- 11477419 TI - Viktor Hamburger 1900-2001. PMID- 11477421 TI - CRMP-2 induces axons in cultured hippocampal neurons. AB - In cultured hippocampal neurons, one axon and several dendrites differentiate from a common immature process. Here we found that CRMP-2/TOAD-64/Ulip2/DRP-2 (refs. 2-4) level was higher in growing axons of cultured hippocampal neurons, that overexpression of CRMP-2 in the cells led to the formation of supernumerary axons and that expression of truncated CRMP-2 mutants suppressed the formation of primary axon in a dominant-negative manner. Thus, CRMP-2 seems to be critical in axon induction in hippocampal neurons, thereby establishing and maintaining neuronal polarity. PMID- 11477422 TI - Amodal completion of acoustic signals by a nonhuman primate. AB - Evidence of amodal completion exists for both visual and auditory stimuli in humans. The importance of this mechanism in forming stable representations of sensory information suggests that it may be common among multiple modalities and species. Here we show that a species of nonhuman primate amodally completes biologically meaningful acoustic stimuli, which provides evidence that the neural mechanism mediating this aspect of auditory perception is shared among primates, and perhaps other taxonomic groups as well. PMID- 11477423 TI - Category specificity in reading and writing: the case of number words. AB - In standard models, word meanings contribute to reading words aloud and writing them to dictation. It is known that categories of knowledge and the associated word meanings can be spared or impaired selectively, but it has not been possible to demonstrate that category-specific effects apply to reading and writing. Here we report the case of a neurodegenerative patient with selectively spared numerical abilities whose brain damage left him able to read and write only number words. PMID- 11477424 TI - Glutamate regulates actin-based motility in axonal filopodia. AB - The dynamics of axonal arbors during synaptogenesis and their plasticity in the adult nervous system remain poorly understood. Axonal filopodia, which emerge from the shaft of axonal branches and contain small synaptic vesicle clusters, initiate synaptic formation. We found that the movement of axonal filopodia is strongly inhibited by the neurotransmitter glutamate. This inhibitory effect is local, requires extracellular Ca2+, and can be blocked by CNQX treatment but not by NMDA, implicating axonal AMPA/kainate glutamate receptors. Transport and exo endocytic recycling of synaptic vesicle packages in filopodia are not affected. These results reveal that the effect of glutamate on axonal filopodia is similar to its previously described effect on dendritic spines. Our results raise the possibility that axonal ionotropic glutamate receptors are also involved in synaptic plasticity in the adult nervous system. PMID- 11477425 TI - Molecular determinants of NMDA receptor internalization. AB - Although synaptic AMPA receptors have been shown to rapidly internalize, synaptic NMDA receptors are reported to be static. It is not certain whether NMDA receptor stability at synaptic sites is an inherent property of the receptor, or is due to stabilization by scaffolding proteins. In this study, we demonstrate that NMDA receptors are internalized in both heterologous cells and neurons, and we define an internalization motif, YEKL, on the distal C-terminus of NR2B. In addition, we show that the synaptic protein PSD-95 inhibits NR2B-mediated internalization, and that deletion of the PDZ-binding domain of NR2B increases internalization in neurons. This suggests an involvement for PSD-95 in NMDA receptor regulation and an explanation for NMDA receptor stability at synaptic sites. PMID- 11477426 TI - Spontaneous astrocytic Ca2+ oscillations in situ drive NMDAR-mediated neuronal excitation. AB - Astrocytes respond to chemical, electrical and mechanical stimuli with transient increases in intracellular calcium concentration ([Ca2+]i). We now show that astrocytes in situ display intrinsic [Ca2+]i oscillations that are not driven by neuronal activity. These spontaneous astrocytic oscillations can propagate as waves to neighboring astrocytes and trigger slowly decaying NMDA receptor mediated inward currents in neurons located along the wave path. These findings show that astrocytes in situ can act as a primary source for generating neuronal activity in the mammalian central nervous system. PMID- 11477427 TI - The consolidation of new but not reactivated memory requires hippocampal C/EBPbeta. AB - Long-term memory formation consists of multiple phases. A new memory is initially labile and sensitive to disruption by a variety of interfering events or agents. To become stable, this new memory undergoes a process known as consolidation, which, in the case of declarative memories, occurs within the medial temporal lobes and requires gene expression. When recalled, memories re-enter a new phase of vulnerability and seem to require a reconsolidation process in order to be maintained. Here we show that consolidation but not reconsolidation of inhibitory avoidance memory requires the expression of the transcription factor CCAAT enhancer binding protein beta (C/EBPbeta) in the hippocampus. Furthermore, in the same region, de novo protein synthesis is not essential for memory reconsolidation. C/EBPbeta is an evolutionarily conserved genetic marker that has a selective role in the consolidation of new but not reactivated memories in the hippocampus. PMID- 11477428 TI - Natural signal statistics and sensory gain control. AB - We describe a form of nonlinear decomposition that is well-suited for efficient encoding of natural signals. Signals are initially decomposed using a bank of linear filters. Each filter response is then rectified and divided by a weighted sum of rectified responses of neighboring filters. We show that this decomposition, with parameters optimized for the statistics of a generic ensemble of natural images or sounds, provides a good characterization of the nonlinear response properties of typical neurons in primary visual cortex or auditory nerve, respectively. These results suggest that nonlinear response properties of sensory neurons are not an accident of biological implementation, but have an important functional role. PMID- 11477429 TI - Efficient computation and cue integration with noisy population codes. AB - The brain represents sensory and motor variables through the activity of large populations of neurons. It is not understood how the nervous system computes with these population codes, given that individual neurons are noisy and thus unreliable. We focus here on two general types of computation, function approximation and cue integration, as these are powerful enough to handle a range of tasks, including sensorimotor transformations, feature extraction in sensory systems and multisensory integration. We demonstrate that a particular class of neural networks, basis function networks with multidimensional attractors, can perform both types of computation optimally with noisy neurons. Moreover, neurons in the intermediate layers of our model show response properties similar to those observed in several multimodal cortical areas. Thus, basis function networks with multidimensional attractors may be used by the brain to compute efficiently with population codes. PMID- 11477430 TI - Complex objects are represented in macaque inferotemporal cortex by the combination of feature columns. AB - Intrinsic signal imaging from inferotemporal (IT) cortex, a visual area essential for object perception and recognition, revealed that visually presented objects activated patches in a distributed manner. When visual features of these objects were partially removed, the simplified stimuli activated only a subset of the patches elicited by the originals. This result, in conjunction with extracellular recording, suggests that an object is represented by a combination of cortical columns, each of which represents a visual feature (feature column). Simplification of an object occasionally caused the appearance of columns that were not active when viewing the more complex form. Thus, not all the columns related to a particular feature were necessarily activated by the original objects. Taken together, these results suggest that objects may be represented not only by simply combining feature columns but also by using a variety of combinations of active and inactive columns for individual features. PMID- 11477431 TI - Human pitch perception is reflected in the timing of stimulus-related cortical activity. AB - 'Pitch' refers to a sound's subjective highness or lowness, as distinct from 'frequency,' which refers to a sound's physical structure. In speech, music and other natural contexts, complex tones are often perceived with a single pitch. Using whole-head magnetoencephalography (MEG) and stimuli that dissociate pitch from frequency, we studied cortical dynamics in normal individuals who extracted different pitches from the same tone complexes. Whereas all subjects showed similar spatial distributions in the magnitude of their brain responses to the stimuli, subjects who heard different pitches exhibited contrasting temporal patterns of brain activity in their right but not their left hemispheres. These data demonstrate a specific relationship between pitch perception and the timing (phase) of dynamic patterns of cortical activity. PMID- 11477432 TI - Differential responses in the fusiform region to same-race and other-race faces. AB - Many studies have shown that people remember faces of their own race better than faces of other races. We investigated the neural substrates of same-race memory superiority using functional MRI (fMRI). European-American (EA) and African American (AA) males underwent fMRI while they viewed photographs of AA males, EA males and objects under intentional encoding conditions. Recognition memory was superior for same-race versus other-race faces. Individually defined areas in the fusiform region that responded preferentially to faces had greater response to same-race versus other-race faces. Across both groups, memory differences between same-race and other-race faces correlated with activation in left fusiform cortex and right parahippocampal and hippocampal areas. These results suggest that differential activation in fusiform regions contributes to same-race memory superiority. PMID- 11477433 TI - Megadose transplantation of purified peripheral blood CD34(+) progenitor cells from HLA-mismatched parental donors in children. AB - We performed HLA-mismatched stem cell transplantation with megadoses of purified positively selected mobilized peripheral blood CD34(+) progenitor cells (PBPC) from related adult donors in 39 children lacking an otherwise suitable donor. The patients received a mean number of 20.7 +/- 9.8 x 10(6)/kg purified CD34(+) and a mean number of 15.5 +/- 20.4 x 10(3)/kg CD3(+) T lymphocytes. The first seven patients received short term (<4 weeks) GVHD prophylaxis with cyclosporin A, whereas in all the following 32 patients no GVHD prophylaxis was used. In 38 evaluable patients, five patients experienced primary acute GVHD grade I and one patient grade II. In 32 patients, no signs of primary GVHD were seen and GVHD only occurred after T cell add backs. T cell reconstitution was more rapid if the number of transplanted CD34(+) cells exceeded 20 x 10(6)/kg. Of the 39 patients, 15 are alive and well, 13 died due to relapse and 10 transplant-related deaths occurred. We conclude that the HLA barrier can be overcome by transplantation of megadoses of highly purified mismatched CD34(+) stem cells. GVHD can be prevented without pharmacological immunosuppression by the efficient T cell depletion associated with the CD34(+) positive selection procedure. This approach offers a promising therapeutic option for every child without an otherwise suitable donor. PMID- 11477434 TI - Allogeneic bone marrow transplantation in the developing world: experience from a center in India. AB - We describe our experience of setting up an allogeneic BMT program at the Christian Medical College Hospital, Vellore over a period of 13 years, from October 1986 to December 1999. Two hundred and twenty-one transplants were performed during this period in 214 patients, with seven patients undergoing second transplants. Indication for BMT were thalassemia major - 106 (48%), CML - 30, AML - 35, ALL - 10, SAA - 22, MDS - six and six for other miscellaneous disorders. The mean age of this patient cohort was 15.6 years (range 2-52). Graft versus-host disease of grades III and IV was seen in 36 patients (17%) and this was the primary cause of death in 20 patients (9.2%). All patients and donors were CMV IgG positive. Sepsis was the primary cause of death in 16 patients (7.4%), 10 bacterial, four fungal and two viral. One hundred and ten of this series of patients are alive and disease free (50%) with a median follow-up of 24 months (range 2-116). These results are comparable to those achieved for patients with similar disease status in transplant units in the Western world and cost a mean of US$15 000. PMID- 11477435 TI - Factors affecting neutrophil and platelet reconstitution following T cell depleted bone marrow transplantation: differential effects of growth factor type and role of CD34(+) cell dose. AB - We have performed univariate and multivariate analysis to determine the factors that affect the kinetics of neutrophil and platelet recovery in 546 recipients of T cell-depleted (TCD) marrow allografts. All patients received marrow depleted of mature CD3(+) T cells by complement-mediated lysis using T(10)B(9)-1A3 (n = 489) or Muromonab-Orthoclone OKT3 (n = 57) monoclonal antibodies. Neutrophil engraftment to 0.5 x 10(9)/1 and platelet engraftment to 20 x 10(9)/l were assessed as endpoints. Factors significantly affecting neutrophil or platelet engraftment in the univariate analysis included patient age, T cell dose, anti thymocyte globulin use, gender, diagnosis at transplant, CMV serostatus, HLA mismatch, CD34 cell dose (n = 249), and growth factor use and type. These variables were included in the multivariate Cox proportional hazards regression model. The results showed that a faster rate of neutrophil engraftment was independently associated with CD34(+) cell dose > or = 5 x 10(6)/kg and most strongly with growth factor administration. Faster platelet engraftment was associated with transplantation for chronic leukemia, CD34(+) cell dose > or = 2 x 10(6)/kg, an HLA matched related donor, and the absence of growth factor use. G CSF had a higher relative risk (RR) of enhancing neutrophil engraftment than GM CSF and significantly delayed platelet engraftment. The combined use of G-CSF + GM-CSF was similar to G-CSF alone. The enhancing effect of G-CSF for neutrophil recovery was most striking for patients who engrafted to 0.5 x 10(9)/1 at or before day 12 (RR = 9.5, P < 0.0001) compared to patients who received no growth factor. Conversely, the delaying effect of G-CSF on platelet engraftment was strongest for patients engrafting on or before day 25 (RR = 0.4, P = 0.0004). Of the independent variables affecting engraftment kinetics in recipients of TCD marrow allografts only growth factor, and to a limited extent, CD34(+) cell dose can be controlled by the clinician. A higher CD34(+) cell dose enhances the rate of both neutrophil and platelet engraftment whereas for G-CSF the benefits of myeloid growth factor use in enhancing neutrophil recovery may be partly offset by a delay in platelet engraftment. PMID- 11477436 TI - Allogeneic stem cell transplantation reduces disease progression compared to autologous transplantation in patients with multiple myeloma. AB - This study compares the probability of disease progression, progression-free survival, and overall survival between patients undergoing an allogeneic or autologous transplant for multiple myeloma using an identical preparative regimen. Patients received a preparative regimen of TBI, busulfan, and cyclophosphamide followed by an allogeneic or autologous transplant. In the allogeneic group (n = 21), six patients received bone marrow and 15 received G CSF mobilized PBSC; all autologous patients (n = 35) received PBSC mobilized with cyclophosphamide and G-CSF. Allogeneic donors were HLA-identical (n = 20) or one antigen mismatched (n = 1) siblings. Graft-versus-host disease (GVHD) prophylaxis consisted of tacrolimus (n = 10), tacrolimus/methotrexate (n = 6), cyclosporine/methotrexate (n = 4), or cyclosporine (n = 1). The groups were evenly matched for gender, pretransplant therapy, disease status at time of transplant, myeloma subtype, and time from diagnosis to transplant. The median age was significantly lower in the allogeneic group (48 vs 55 years, P < 0.01). In the allogeneic group the probabilities of developing acute GVHD grade II-IV and chronic GVHD were 55% and 82%, respectively. The Kaplan-Meier probability of disease progression was significantly lower in the allogeneic group (11% vs 64%, P < 0.001) compared to the autologous group. Although progression-free (60% vs 30%, P = 0.19) and overall survival at 2 years (60% vs 42%, P = 0.39) favored the allogeneic group, this did not reach statistical significance. Within the allogeneic transplant group, patients age 50 years or under had a 3-year overall survival significantly higher when compared to older patients (79% vs 29%, P = 0.03). Using identical preparative regimens, allogeneic transplantation reduced disease progression compared to autologous transplantation for myeloma. This suggests that allogeneic transplantation induces a graft-versus-myeloma (GVM) effect. PMID- 11477437 TI - A comparison of chimerism and minimal residual disease between four different allogeneic transplantation methods in patients with chronic myelogenous leukemia in first chronic phase. AB - The detection of chimerism, residual molecular and cytogenetic disease following transplantation of peripheral blood stem cells (PBSCT) with a nonmyeloablative conditioning (n = 9) and the transplantation of highly purified CD34(+) stem cells (CD34(+) PBSCT) (n = 16) were compared to unmanipulated bone marrow transplantation (BMT) (n = 69) and unmanipulated PBSCT (n = 50) after myeloablative conditioning in patients with first chronic phase of chronic myelogenous leukemia (CML) (n = 137), second chronic phase of CML (n = 4), acute lymphoblastic leukemia (n = 2) and acute myeloid leukemia (n = 1). A molecular relapse (MR) as defined by two consecutive positive polymerase chain reaction assays for the detection of M-bcr-abl transcripts (n = 141) and cbfbeta-myh11 transcripts (n = 1) in a 4-week interval was found in 10 of 16 patients (63%) after CD34(+) PBSCT, and in 27 of 69 patients (39%) after BMT, whereas only three of 50 patients (6%) after PBSCT (P < 0.001) and one of eight patients (13%) after PBSCT with reduced conditioning suffered from a MR. A cytogenetic relapse occurred in five of 16 patients (31%) after CD34(+)PBSCT and 21 of 69 patients (30%) after BMT (NS) compared to two of 50 patients (4%) after PBSCT and none of the eight patients after PBSCT with reduced conditioning (P < 0.05). The lowest treatment-related mortality was seen in the 16 patients after CD34(+) PBSCT, who are all currently alive with a median follow-up of 15 months, whereas the survival rate for BMT, PBSCT and PBSCT with reduced conditioning were 65%, 63% and 58%, respectively. Multivariate analysis including all potential influential factors of post-transplant residual disease recurrence showed that patients after CD34(+) PBSCT had a significantly higher risk (two times) to develop a MR than patients after BMT (P < 0.03), whereas patients after unmanipulated PBSCT had a significant lower risk (eight times) for the occurrence of a MR post transplant (P < 0.001). Patients after BMT and CD34(+) PBSCT had the lowest rates of complete chimerism (CC) at 3 months after transplant. Only five of nine patients (55%) after CD34(+) PBSCT and 19 of 33 patients (58%) after BMT achieved CC compared to 19 of 22 (86%) patients after PBSCT and seven of eight (88%) patients after PBSCT with reduced conditioning (P < 0.05). PMID- 11477438 TI - Degree of fatness after allogeneic BMT for childhood leukaemia or lymphoma. AB - Excess fatness is frequent after childhood ALL treated without BMT. We measured the whole-body percent fat by dual-energy X-ray absorptiometry and the body-mass index (weight/height(2) (kg/m(2)), BMI) in 25 survivors of childhood leukaemia or lymphoma (21 with ALL) who had received TBI and allogeneic BMT a median of 8 years ago (range 4-13). Adjusted for sex and age, the mean BMI was slightly but significantly reduced (0.4 s.d. below predicted) and the whole-body percent fat was significantly increased compared with healthy controls (1.1 s.d. above predicted). Eleven of 25 patients had a percent fat above the 90 percentile of the reference values, which indicates excess fatness. Adjusted for sex and age, a higher percent fat was related to additional cranial irradiation. Controlled for this, the whole-body percent fat seemed to be unrelated to age at BMT, length of follow-up, and previous chemotherapy. Compared with untransplanted ALL survivors treated with cranial irradiation, BMT survivors had significantly reduced BMI but similar whole body percent fat. BMI was a poor measure of body fatness in these patients. In conclusion, survivors of BMT for childhood leukaemia or lymphoma are adipose and slightly underweight and consequently have a substantially reduced lean body mass. PMID- 11477439 TI - Thiotepa, busulfan, cyclophosphamide (TBC) and autologous hematopoietic transplantation: an intensive regimen for the treatment of multiple myeloma. AB - The study was designed to evaluate the efficacy and safety of an intensive, tri alkylator conditioning regimen, consisting of thiotepa, busulfan and cyclophosphamide (TBC), prior to autologous hematopoietic cell transplantation in patients with multiple myeloma (MM) and to analyze factors associated with outcome. One hundred and twenty patients with MM received high-dose chemotherapy with TBC followed by autologous bone marrow (n = 24) or peripheral blood stem cell (PBSC) transplantation (n = 96). Fifty-four patients had chemosensitive disease and 66 had refractory disease at the time of transplantation. The overall response rate was 81% and the complete remission (CR) rate was 26%. Patients with chemosensitive disease had a CR rate of 52% vs 5% for patients with refractory disease. Multivariable analysis determined disease status at transplant as the factor most likely associated with long survival. Estimated median survival was 48, 35 and 9 months for patients with chemosensitive, primary refractory or disease in refractory relapse, respectively. Short interval from diagnosis to transplant among patients with primary refractory disease and younger age were also favorable prognostic factors for survival. Patients with refractory disease pre-transplant who achieved remission criteria rapidly after treatment had a worse outcome than the slow responders. Treatment-related mortality with the introduction of PBSC and better supportive care was 4.8%. In conclusion, TBC is an effective and relatively well-tolerated intensive conditioning regimen in patients with MM. A more favorable outcome was observed in patients with chemosensitive disease and with early treatment for primary refractory disease. TBC merits further study in these subgroups and comparison with alternative regimens in prospective studies is warranted. PMID- 11477440 TI - Double reinforcement with fludarabine/high-dose cytarabine enhances the impact of autologous stem cell transplantation in acute myeloid leukemia patients. AB - Reinforced chemotherapy based on a double high-dose consolidation regimen could be a different way to enhance in vivo purging prior to autologous stem cell transplantation (auto-SCT) in acute myeloid leukemia (AML). We investigated the impact on outcome of auto-SCT after two different strategies of early intensification performed after an identical induction regimen in adult patients with AML. Between January 1993 and December 1998, 140 consecutive AML patients were enrolled in a program consisting of an identical anthracycline-based induction (ICE) and two different consolidation regimens: one cycle, cytarabine based (single-NOVIA: 91 patients); two cycles, fludarabine-based (double-FLAN: 49 patients). Seventy out of 91 patients received single-NOVIA consolidation: 60 underwent a transplantation procedure (allogeneic bone marrow transplantation (allo-BMT):16 patients; auto-SCT: 44). Thirty-five out of 49 patients received double-FLAN consolidation: 31 underwent a transplantation procedure (allo-BMT: 10; auto-SCT: 21). The double consolidation regimen was well-tolerated with only minor side-effects. Median follow-up observation time for surviving patients was 38 months (range, 17-71) for the double-FLAN consolidation group and 70 months (range: 48-93) for the single-NOVIA consolidation group. Among the patients who received auto-SCT, the double consolidation strategy produced a superior disease free survival curve at 36 months (78.6% (95%CI: 59.4-97.8) vs 47.7% (95%CI: 33 62.4)) compared with the single-NOVIA group. This difference was confirmed when the patients were analyzed for intention to treat (P = 0.04). In addition, the double-FLAN consolidation group showed a superior overall survival and lower relapse rate (P = 0.02). We conclude that the double-FLAN reinforcement strategy is safe and enhances the clinical impact of auto-SCT for AML patients in first complete remission. It may provide specific clinical benefit for patients undergoing auto-SCT. PMID- 11477441 TI - A comparison of toxicity following two different doses of cyclophosphamide for mobilization of peripheral blood progenitor cells in 116 multiple myeloma patients. AB - High-dose cyclophosphamide (HDC) has been shown to be an effective regimen for collecting PBPC in multiple myeloma (MM) patients, but the optimal dose to be used remains controversial. Two historical cohorts of MM patients who received G- or GM-CSF and HDC at the dose of either 7 g/m(2) (HDC7, n = 74) or 4 g/m (HDC4, n = 42) were compared. As patients in the HDC4 group were more likely to have received G-CSF than GM-CSF (P < 10(-3)) and fewer previous alkylating agents (P = 0.004), multivariate logistic regression analysis was performed. In the HDC4 group, patients had a shorter median duration of neutropenia (P < 10(-4)), fewer RBC (P < 10(-3)) and platelet transfusions (P < 10(-3)) with fewer patients with platelets <20 x 10(9)/l (P = 0.004). Moreover, fewer febrile episodes (P < 10( 3)) and less need of intravenous antibiotics (P < 10(-3)) were found in the HDC4 group. No statistical difference was observed with regard to CD34(+) cell collection efficiency. Thus, the use of HDC at the dose of 4 g/m(2) for the collection of PBPC in MM patients decreases hematological and extrahematological toxicity with an equivalent CD34(+) cell collection efficiency. PMID- 11477442 TI - Prognostic importance of the axillary lymph node ratio in autologous transplantation for high-risk stage II/III breast cancer. AB - The role of autologous peripheral blood progenitor cell (PBPC) transplantation for high-risk stage II/III breast cancer remains controversial. New prognostic indicators defining subsets of patients who may benefit from autologous PBPC transplantation would be clinically useful. The axillary lymph node ratio, defined by the total number of axillary nodes involved with cancer divided by the number of axillary nodes surgically sampled, has been reported to be of potential prognostic importance in transplantation for high-risk, stage II/III breast cancer. We therefore retrospectively reviewed 111 women with high-risk, stage II/III breast cancer with at least four positive axillary lymph nodes undergoing autologous PBPC transplantation from 1991 to June 1999. None of the patients had received prior radiotherapy and all had completed one, and only one, course of at least three cycles of adjuvant chemotherapy. The median number of axillary nodes sampled was 20 (range 6-40) and the median number of positive axillary nodes was 12 (range 4-35). The median node ratio, dividing the number of positive nodes by the number of sampled nodes, was 0.68. Event-free survival was strongly influenced by node ratio. Patients having a node ratio of < 0.7 had a 5-year event-free survival of 68%, vs those with a node ratio of > or = 0.7 with a 5 year event-free survival of 46% (P = 0.03). Forty percent of patients with a high node ratio have relapsed vs 20% with a low node ratio (P = 0.02). Multivariate analysis revealed that positive estrogen receptor status and a node ratio of < 0.7 were independent factors related to better event-free survival (P = 0.0001 and P = 0.004, respectively). We conclude that patients having a node ratio of < 0.7 have a significantly better prognosis following autologous PBPC transplantation than do patients with a ratio > or = 0.7. PMID- 11477443 TI - HER-2 expression is a prognostic factor in patients with metastatic breast cancer treated with a combination of high-dose cyclophosphamide, mitoxantrone, paclitaxel and autologous blood stem cell support. AB - The expression levels of a circulating extracellular domain of HER-2 can be detected in the plasma and serum of patients with metastatic breast cancer using an enzyme immunoassay (ELISA) method. In this study, we evaluated the clinical significance of high and low levels of HER-2 in the plasma of 46 patients with metastatic breast cancer enrolled in a clinical trial of high-dose chemotherapy (HDCT) using cyclophosphamide, mitoxantrone, and paclitaxel with autologous stem cell transplantation (ASCT). Using 2500 U/ml as the cut-point, 20 patients (46%) had elevated HER-2 levels (HER-2 positive). Our results suggest that patients with metastatic breast cancer and high soluble plasma HER-2 have a significantly poorer overall (OS) and progression-free survival (PFS) following high-dose chemotherapy with paclitaxel and ASCT. The median OS of patients with low levels of HER-2 was significantly longer (P < 0.01) than the median OS of patients with high levels of HER-2 (29.8 months vs 15.9 months). PFS was also significantly longer (P < 0.01) for patients who were HER-2-negative, than for patients who were HER-2-positive (13.0 vs 8.6 months). Univariate analysis showed that patients with liver or lung metastases had significantly reduced OS and PFS. Patients with metastases to two or more sites also had a significantly reduced time to disease progression, but not OS. In multivariable analysis, lung metastases contributed along with HER-2-positive status to determine a group of patients with significantly poorer OS. However, HER-2-positive status remained the only independent predictor of PFS. PMID- 11477444 TI - Oral busulfan pharmacokinetics and engraftment in children with Hurler syndrome and other inherited metabolic storage diseases undergoing hematopoietic cell transplantation. AB - Allogeneic hematopoietic cell transplantation (HCT) is the only treatment for selected inherited metabolic storage diseases (IMSD); a significant shortcoming is failure to achieve donor-derived engraftment. This study was undertaken to determine whether busulfan pharmacokinetics (BU PK) are altered in children with IMSD and whether BU concentrations are important in achieving engraftment. BU samples were obtained from 39 IMSD children, including 20 children with Hurler syndrome, undergoing HCT. Patients received oral BU (40 mg/m(2)/dose x 8 doses), cyclophosphamide (60 mg/kg/day x 2 doses) and TBI (750 cGy in one fraction) as a preparative regimen. Median (range) oral clearance corrected for bioavailability (Cl/F in ml/min/kg), area under the curve (AUC in ng min/ml) and BU plasma concentration (Cp in ng/ml) with the fourth dose were 5.2 (2.1-11.4), 318 294 (112 893-640 995) and 950 (314-1780), respectively. Children < 3 years of age had lower AUC and Cp but higher Cl/F (P < or = 0.03). BU Cp (P = 0.06) or marrow cell dose (P = 0.32) was not different in Hurler syndrome compared to other IMSD. A median BU Cp of 959 and 831 ng/ml was achieved in children with full and failed early engraftment, respectively. There was no difference in early and late engraftment between children with Hurler and other IMSD. In conclusion, we found no significant association between engraftment, marrow cell dose and BU exposure when combined with CY and TBI in children with IMSD. PMID- 11477445 TI - MC1288, a vitamin D analog, prevents acute graft-versus-host disease in rat bone marrow transplantation. AB - The major obstacle to successful bone marrow transplantation (BMT) is graft versus-host disease (GVHD). Vitamin D analogs have shown their efficacy in solid organ transplantation. The purpose of this study was to investigate the suitability of a novel vitamin D analog, MC1288, in the prevention of acute GVHD in a rat BMT model. Allogeneic BMT were performed from Lewis to BN rats (n = 18). The animals were divided into four groups: an untreated control group, MC1288, cyclosporin A (CsA), and MC1288 + CsA-treated groups. Rats were harvested for histology and immunohistochemistry on day 20 after BMT. Histological changes for GVHD in liver, skin, and spleen were scored. Positivity in immunostaining was quantified as the number of positive cells/high power field. Treatment with MC1288 decreased clinical signs of GVHD compared with untreated or CsA-treated rats. Histological manifestations of GVHD, expressed as mean total increment, were significantly lower (1.4 +/- 0.5) in MC1288 than in untreated (5.0 +/- 1.6) or CsA (3.5 +/- 1.0) groups. Combining MC1288 and CsA further improved histology (1.1 +/- 0.6). The expression of CD4, CD8, MHC class II, interleukin-2 receptor, nitric oxide 2, and NKR-P1A (NK cells) positivity was significantly decreased in the liver and skin of BMT rats by MC1288. MC1288 was effective in preventing clinical and histological signs and symptoms of GVHD. This novel vitamin D analog could be used as an immunomodulating agent in BMT. PMID- 11477446 TI - In vitro generation of cytotoxic effectors activated by interleukin 2 (IL-2): comparison of autologous peripheral blood stem cells (PBSC) from adults and children. AB - Previous studies demonstrated that ex vivo IL-2- activated PBSC could generate cytotoxic effectors without impairing haematopoietic reconstitution. Clinical experience and previous studies indicated that children with solid tumours could benefit from high-dose chemotherapy with immune modulation. We studied the generation of cytotoxic effectors from growth-factor +/- chemotherapy-mobilised PBSC from 10 patients (five adults and five children) with different solid tumours. Cells were placed in culture in serum-free culture medium supplemented with IL-2 1000 U/ml +/- IL-12 for 1, 2, 4 or 7 days. Anti-tumour cytotoxicity was tested against K562, Daudi and two neuroblastoma cell lines (Gau, NB91). Cultured adult PBSC in the presence of IL-2 (1000 U/ml) showed marked cytotoxicity against all the cell lines tested from day 1. At day 2, with an E:T ratio of 25:1, cytotoxicity was 53% +/- 10.4, 63.2% +/- 23.8, 38% +/- 9.1, and 39% +/- 15.7 against K562, Daudi, Gau and NB91, respectively. Cytotoxic activity of child PBSC was significantly lower (P < 0.05) and was displayed after longer culture times (day 4). No difference was found in the phenotype analysis of lymphoid subsets before and after IL-2 activation between adult and child PBSC. Haematological properties of the graft were not significantly impaired by IL-2 activation. PMID- 11477447 TI - Cytomegalovirus pneumonia in adult autologous blood and marrow transplant recipients. AB - CMV pneumonia is a major cause of morbidity and mortality among allogeneic BMT recipients. To assess the frequency, timing, risk factors and response to therapy of CMV pneumonia among autologous BMT recipients, we reviewed our experience with 795 patients. Sixteen (2%) patients were diagnosed with CMV pneumonia. The frequency was higher among patients who were seropositive than those who were seronegative (3.3% vs 0%, P = 0.008). Among seropositive patients, the frequency was higher among patients with hematological malignancies than patients with solid tumors (5.0 % vs 1.0%, P = 0.019). Eleven cases occurred <30 days, and five cases occurred >100 days post transplant. The overall CMV pneumonia-related mortality rate was 31%. Seven (78%) of nine patients treated with ganciclovir and IVIG prior to respiratory failure survived; neither of two patients treated after respiratory failure survived. Four of five (80%) untreated patients survived. In conclusion, CMV is a not infrequent cause of pneumonia among autologous BMT recipients. Risk factors include CMV seropositivity and an underlying hematological malignancy. A favorable response hinges on the prompt initiation of therapy. The survival of 25% of the patients without antiviral therapy suggests that the isolation of CMV from a BAL specimen occasionally reflects oropharyngeal contamination or that CMV pneumonia may sometimes be self-limited in more immunocompetent autologous BMT recipients. PMID- 11477448 TI - Successful umbilical cord blood transplantation from an unrelated donor for a patient with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis. AB - We report a case of a 5-year-old girl with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) who underwent cord blood (CB) stem cell transplantation (CBSCT) from an unrelated donor. The patient presented with persistent high-grade fever and hepatosplenomegaly. Because the disease was refractory to immunochemotherapy according to the HLH94 protocol, she received 2.0 x 10(7) CB nucleated cells/kg body weight (BW) after conditioning with BU/CY/etoposide. No acute GVHD developed, using FK506 for prophylaxis. The neutrophil count reached >0.5 x 10(9)/l by day 21 and the platelet count reached >50 x 10(9)/l by day 84. The patient recovered well with sequelae of neurological deficits more than 10 months after receiving CBSCT, without showing evidence of HLH or chronic GVHD. Real-time PCR proved applicable for estimation of the EBV load in PBMC of the patient. We conclude that CBSCT may be indicated for some cases of refractory EBV-HLH, who have no HLA-matched siblings and are therefore dependent on unrelated marrow donors. PMID- 11477449 TI - Development of spontaneous factor VIII inhibitor in association with acute graft versus-host disease. AB - A 57-year-old female with recurrent AML underwent a T cell-depleted (TCD) bone marrow (BM) plus TCD and CD34-selected peripheral blood stem cell (PBSC) transplant. Eleven weeks post transplantation, the patient developed acute graft versus-host disease (GVHD) manifested by rash and elevated liver enzymes. Concurrently, the patient presented with a bleeding diathesis and a left forearm hematoma due to the development of a spontaneous factor VIII inhibitor. She was treated with human recombinant factor VIII and intravenous methylprednisolone. Subsequently she was managed with a prednisone taper leading to resolution of the GVHD, as well as the spontaneous factor VIII inhibitor. Bone marrow transplant related spontaneous factor VIII inhibitor has previously been reported in association with one patient with chronic GVHD. To our knowledge this is the first report of spontaneous factor VIII inhibitor associated with acute GVHD. PMID- 11477450 TI - Antithymocyte-associated reticulonodular pneumonitis during a conditioning regimen of reduced intensity for genoidentical bone marrow transplantation. PMID- 11477451 TI - Safety profile of mycophenolate mofetil: a response. PMID- 11477452 TI - Immunotherapy of established tumors in mice by intratumoral injection of an adenovirus vector harboring the human IL-2 cDNA: induction of CD8(+) T-cell immunity and NK activity. AB - Intratumoral (i.t.) injections of an adenovirus encoding the human interleukin-2 (IL-2) under the control of the RSV (Ad-pRSV-IL-2) or CMV (Ad-pCMV-IL-2) promoter were performed in established mastocytoma P815 tumors in B6D2 mice. Both early and long-term survival were found increased in mice treated with Ad-pCMV-IL-2 as compared with those obtained with Ad-pRSV-IL-2: tumor regression was observed in 30--50% of mice for the former and 5--15% for the latter. Difference in efficacy between the two vectors was directly correlated to the amount of IL-2 produced i.t. between 24 and 48 hours postinjection, which reached 10--20 ng/tumor for Ad pCMV-IL-2 and 0.3--0.5 ng/tumor for Ad-pRSV-IL-2. In both cases, expression in the tumor was clearly detectable for a period of 7--10 days postinjection. Serum IL-2 was not detectable in mice treated with Ad-pRSV-IL-2, whereas expression peaked at a total of 1--2 ng at 24 hours but declined very rapidly in the Ad-pCMV IL-2-treated group. Constant production of IL-2 inside the tumor was necessary for successful therapy because i.t. injections of recombinant IL-2 at levels up to 1 microg for five consecutive days did not lead to antitumoral activity. Evidence of induced systemic immunity following Ad-pCMV-IL-2 injections was obtained from rechallenge experiments in which tumor-free mice after treatment rejected a subsequent contralateral injection of a lethal dose of P815 tumor cells and from the observation that regression of nontreated tumors occurred in animals bearing bilateral tumors that were treated i.t. in a single tumor with Ad pCMV-IL-2. P815-specific cytotoxic T lymphocytes (CTL) were found specifically in spleen cells from cured mice or rechallenged mice but not in control mice. Interestingly, limiting dilution analysis of anti-P815 CTL precursor (CTLp) frequency revealed a significant increase in mice cured of their tumor as compared to that obtained in naive mice or control mice treated or not with Ad-IL 2 but whose tumor was growing. In vivo depletion of T-cell subsets, as well as natural killer cells at the time of i.t. injections with Ad-pCMV-IL-2, demonstrated that both CD8(+) T cells and natural killer cells, but not CD4(+) T cells, were required for successful therapy. Finally, mice preimmunized with Ad null viruses were severely compromised in their capacity to eradicate established P815 tumors after Ad-pCMV-IL-2 therapy, at least when neutralizing antibody titers reached a critical level. PMID- 11477453 TI - The cytotoxic effect of E1B 55-kDa mutant adenovirus on human hepatocellular carcinoma cell lines. AB - It has been suggested the E1B 55 kDa mutant adenovirus dl1520 can selectively kill p53-deficient human tumor cells. In this study, we examined the cytotoxic effect of dl1520 on nine human hepatocellular carcinoma (HCC) cell lines with different p53 genetic and functional status. The results showed that HCC cell lines with deleted or mutant p53 gene and reduced p53 transcriptional activities were more susceptible to dl1520-induced cytolysis. Hep3B (p53-null) and HepG2 (p53-wt) cells were arrested at G2/M phase when cytolysis occurred. Cyclin dependent kinase inhibitor (CDKI) p21(Waf-1/Cip-1) was downregulated 24 hours after dl1520 infection in HepG2 cells and increased when cytolysis occurred. No p21 expression was detected in Hep3B cells. DNA fragmentation was found in both Hep3B and HepG2 cells after dl1520 infection. Bax expression increased in dl1520 infected HepG2 cells but not in Hep3B cells. Notably, three Bax-like proteins, molecular mass around 40 to 80 kDa, accumulated 48 hours after adenovirus infection in Hep3B cells but not in HepG2 cells. These results suggest that the susceptibility of HCC cells to dl1520-induced cytolysis is related to both p53 genotype and functional status, and is mediated by both cell cycle disturbance and apoptosis. PMID- 11477454 TI - Selective gene delivery toward gastric and esophageal adenocarcinoma cells via EpCAM-targeted adenoviral vectors. AB - Application of recombinant adenoviral vectors for cancer gene therapy is currently limited due to lack of specificity for tumor cells. For gastric and esophageal adenocarcinoma, we present here that the relative abundant expression of the primary adenovirus receptor, coxsackie/adenovirus receptor (CAR), on normal epithelium compared to carcinoma favors the transduction of the epithelium. As such, to achieve specific transduction of cancer cells, targeting approaches are required that ablate the binding of the virus to CAR and redirect the virus to tumor-specific receptors. By immunohistochemistry and reverse transcriptase polymerase chain reaction assays, we demonstrate a marked difference in expression of the human epithelial cell adhesion molecule (EpCAM) between normal and (pre)malignant lesions of the stomach and esophagus. Based on this, we explored the feasibility of using EpCAM to achieve gastric and esophageal adenocarcinoma selective gene transfer. Adenoviral vectors redirected to EpCAM using bispecific antibodies against the adenovirus fiber-knob protein and EpCAM specifically infected gastric and esophageal cancer cell lines. Using primary human cells, an improved ratio of tumor transduction over normal epithelium transduction was accomplished by the EpCAM-targeted vectors. This study thus indicates that EpCAM-targeted adenoviral vectors may be useful for gastric and esophageal cancer-specific gene therapy in patients. PMID- 11477455 TI - Cisplatin chemotherapy plus adenoviral p53 gene therapy in EBV-positive and negative nasopharyngeal carcinoma. AB - We have previously shown that the introduction of human recombinant wild-type p53 mediated by an adenoviral vector (Ad5CMV-p53), either alone or delivered in combination with ionizing radiation, was cytotoxic to two nasopharyngeal carcinoma (NPC) cell lines. To further explore the potential therapeutic role for gene therapy, the combination of Ad5CMV-p53 and cisplatin was examined in two NPC cell lines, CNE-1 and C666-1. The C666-1 cells are particularly relevant because they express Epstein-Barr virus latent gene products analogous to human NPC in situ. Cells were infected with 5 pfu/cell of Ad5CMV-p53 or Ad5CMV-beta-gal, followed by exposure to increasing doses of cisplatin. Clonogenic and MTT assays were used to assess the sensitivity of cells to these treatments, and apoptosis was also quantified. The combination of Ad5CMV-p53 and cisplatin resulted in approximately 25% greater cytotoxicity compared to that observed with cisplatin alone in either cell line. Apoptosis was induced in approximately 50% of cells following administration of both Ad5CMV-p53 and cisplatin, but was induced in considerably fewer cells following either treatment alone. The two modalities appeared to interact in an additive manner. Ad5CMV-p53 gene therapy resulted in the expression of biologically active p53 protein, shown by induction of p21(WAF1/CIP1). Cisplatin treatment showed little effect on either p53 or p21(WAF1/CIP1) expression. Therefore, both p53 gene therapy and cisplatin chemotherapy demonstrated cytotoxicity mediated by apoptosis despite the presence of EBV gene products in the C666-1 cells, but it appears that the two modalities induce cytotoxicity by independent pathways. PMID- 11477456 TI - Tricistronic viral vectors co-expressing interleukin-12 (1L-12) and CD80 (B7-1) for the immunotherapy of cancer: preclinical studies in myeloma. AB - Synergy between interleukin-12 (IL-12) and B7-1 (CD80) for cancer immunotherapy has previously been demonstrated in animal models of breast cancer, lymphoma, and multiple myeloma. With a view to human clinical application, tricistronic retroviral and adenovirus vectors co-expressing IL-12 (IL-12p40 plus IL-12p35) and CD80 were constructed by utilizing two internal ribosome entry site (IRES) sequences to link the three cDNAs. A murine stem cell virus (MSCV)-based retroviral vector (MSCV-hIL12.B7) utilized distinct IRES sequences from the encephalomyocarditis virus (EMCV) and the foot-and-mouth disease virus (FMCV), whereas Ad5-based adenovirus vectors contained transcriptional units with two EMCV IRES sequences under the control of murine (AdMh12.B7) or human (AdHh12.B7) cytomegalovirus promoters. AdMh12.B7 was found to consistently direct higher levels of IL-12 and CD80 expression than AdHh12.B7 following infection of a number of human tumor cell lines. In preclinical studies, the human myeloma cell line U266 was infected with MSCV-hIL12.B7 and a resulting clonal cell line, U/MSCV-h12.B7, was generated with stable expression of CD80 and secreting IL-12 at 1 ng/24 h/10(6) cells. By comparison, following AdMh12.B7 infection, 81% of infected U266 cells (U/AdMh12.B7) expressed CD80 and secreted IL-12 at 25-50 ng/24 h/10(6) cells. Both engineered myeloma cell lines stimulated enhanced allogeneic mixed lymphocyte proliferation and provoked increases in cytotoxic T lymphocyte responses and gamma-interferon release from normal donor lymphocytes exposed to parental U266 cells. These results suggest potential clinical utility of AdMh12.B7 in immunotherapy strategies for the treatment of multiple myeloma and other cancers. PMID- 11477457 TI - Antitumor efficacy of a human interleukin-12 expression plasmid demonstrated in a human peripheral blood leukocyte/human lung tumor xenograft SCID mouse model. AB - Genes encoding the p35 and p40 subunits of human interleukin-12 (IL-12) and the bacterial aminoglycoside phosphotransferase were cloned into a mammalian expression plasmid. The resultant plasmid, pCMVIL-12neo, was used to transfect human lung tumor cell lines in vitro. Stably transfected subclones were generated and found to secrete human IL-12 for at least 10 days following a lethal dose of gamma-radiation. The ability of the IL-12--producing tumor cells to promote an antitumor response in vivo was evaluated in SCID mice co-engrafted subcutaneously with human peripheral blood lymphocytes (PBLs) and viable human lung tumor cells (SCID-Winn assay). Using this model system, it was established that IL-12 released locally into tumors by irradiated IL-12--transfected cells activated the human PBL and promoted their ability to suppress tumor development in a dose dependent fashion. PBL subset depletion studies revealed that the antitumor effect promoted by the IL-12--modified cells was dependent on the presence of human CD8(+) T cells and, to a lesser extent, human CD56(+) natural killer cells within the xenograft. We conclude that (a) irradiated human lung tumor cells genetically modified with pCMVIL-12neo secrete bioactive human IL-12 at concentrations sufficient to promote a human lymphocyte-mediated antitumor response in the microenvironment of the xenograft, and (b) that the SCID-Winn assay provides a useful model for the preclinical evaluation of cytokine-based human immunotherapy protocols. PMID- 11477458 TI - Transgenic expression of CD40L and interleukin-2 induces an autologous antitumor immune response in patients with non-Hodgkin's lymphoma. AB - The malignant B cells of non-Hodgkin's lymphoma (B-NHL cells) express peptides derived from tumor-specific antigens such as immunoglobulin idiotypes, and also express major histocompatibility complex antigens. However, they do not express co-stimulatory molecules, which likely contributes to their protection from host antitumor immunity. To stimulate NHL-specific immune responses, we attempted to transfer the human CD40 ligand (hCD40L) gene to B-NHL cells and enhance their co stimulatory potential. We found that an adenoviral vector encoding human CD40L (AdhCD40L) was ineffective at transducing B-NHL cells because these cells lack the coxsackievirus B-adenovirus receptor and alpha(v) integrins. However, preculture of the B-NHL cells with the human embryonic lung fibroblast line, MRC 5, significantly up-regulated expression of integrin alpha(v)beta 3 and markedly increased their susceptibility to adenoviral vector transduction. After prestimulation, transduction with AdhCD40L increased CD40L expression on B-NHL cells from 1.3+/-0.2% to 40.8+/-11.9%. Transduction of control adenoviral vector had no effect. Expression of transgenic human CD40L on these CD40-positive cells was in turn associated with up-regulation of other co-stimulatory molecules including B7-1/-2. Transduced B-NHL cells were now able to stimulate DNA synthesis of autologous T cells. However, the stimulated T cells were unable to recognize unmodified lymphoma cells, a requirement for an effective tumor vaccine. Based on previous results in an animal model, we determined the effects of combined use of B-NHL cells transduced with AdhCD40L and AdhIL2 vectors. The combination enhanced initial T-cell activation and generated autologous T cells capable of specifically recognizing and killing parental (unmodified) B-NHL cells via major histocompatibility complex--restricted cytotoxic T lymphocytes. These findings suggest that the combination of CD40L and IL2 gene-modified B-NHL cells will induce a cytotoxic immune response in vivo directed against unmodified tumor cells. PMID- 11477459 TI - (E)-5-(2-bromovinyl)-2'-deoxyuridine potentiates ganciclovir-mediated cytotoxicity on herpes simplex virus-thymidine kinase--expressing cells. AB - Tumor cells expressing the thymidine kinase gene of the herpes simplex virus (HSV tk) are rendered highly susceptible to the cytotoxic effects of different antiherpes drugs. In an attempt to enhance cytotoxicity of this therapeutic approach in glioma and other tumor cell lines transduced with the HSV-tk gene, we evaluated tumor cell killing following co-administration of two different prodrugs metabolized by HSV-tk, (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU), and ganciclovir (GCV). In 8 of 12 cell lines investigated, addition of BVDU in concentrations showing no cytotoxic effect or only limited cytotoxicity could enhance GCV-mediated cell killing by as much as one order of magnitude. In co cultures consisting of HSV-tk(+) (9L STK) and HSV-tk(-) (9L wild-type) cells, we also observed potentiation of GCV-mediated cytotoxicity in the presence of BVDU, suggesting strongly enhanced bystander cell killing. BVDU is thought to exert its cytotoxic effect through inhibition of thymidylate synthase activity or by incorporation into replicating DNA. Both effects could be observed in all HSV-tk- expressing cells investigated, including cell lines which did not exhibit cytotoxicity after incubation with BVDU. These findings argue against current concepts of BVDU-mediated cytotoxicity in HSV-tk--expressing cells. Taken together, our data suggest that gene therapy utilizing prodrug activating enzymes may be rendered more effective by simultaneous treatment with two different prodrugs metabolized by the same enzyme. PMID- 11477460 TI - Low-dose supplementation with lycopene or beta-carotene does not enhance cell mediated immunity in healthy free-living elderly humans. AB - OBJECTIVE: To assess the effects of supplementation with the carotenoids, beta carotene or lycopene, at levels achievable within a diet rich in fruit and vegetables, on immune function in a group of free-living healthy elderly. DESIGN: A double-blind randomized placebo-controlled trial. T cell subsets and the expression of functionally associated cell surface molecules, quantified by flow cytometry, and lectin-stimulated lymphocyte proliferation, were compared before and following the treatment period. SUBJECTS: Fifty-eight subjects aged over 65 y were recruited; 52 were included in the final analysis. INTERVENTIONS: Participants received one placebo, one lycopene (13.3 mg) or one beta-carotene (8.2 mg) capsule daily for 12 weeks. RESULTS: No significant differences were observed in any of the parameters examined following either lycopene or beta carotene supplementation. CONCLUSIONS: In well-nourished, free-living, healthy elderly individuals, supplementation with relatively low levels of beta-carotene or lycopene is not associated with either a beneficial or detrimental effects on several aspects of cell-mediated immunity. PMID- 11477461 TI - Fruit and vegetables and cardiovascular risk profile: a diet controlled intervention study. AB - OBJECTIVE: To evaluate the effect of fruit and vegetables consumption on markers of risk for cardiovascular disease. DESIGN: Randomised, diet controlled, parallel study. SUBJECTS: Forty-eight apparently healthy (40--60 y) volunteers with a low usual consumption of fruit and vegetables. Forty-seven of them completed the study. INTERVENTIONS: During 4 weeks 24 volunteers consumed a standardised meal, consisting of 500 g/day fruit and vegetables and 200 ml/day fruit juice ('high' group) and 23 volunteers consumed 100 g/day fruit and vegetables ('low' group) with an energy and fat controlled diet. RESULTS: Final total cholesterol was 0.2 (95% CI -0.5--0.03) mmol/l lower in the high group than in the low group (P>0.05). Final fibrinogen and systolic blood pressure were 0.1 (-0.1--0.4) g/l and 2.8 (-2.6--8.1) mmHg higher in the high group than in the low group (P>0.05), respectively. Also, other final serum lipid concentrations, diastolic blood pressure and other haemostatic factors did not differ between both groups. CONCLUSIONS: This was a small randomised well-controlled dietary intervention trial of short duration with a considerable contrast in fruit and vegetable consumption. No effects on serum lipids, blood pressure and haemostatic variables were observed. PMID- 11477462 TI - Single measurement of serum phospholipid fatty acid as a biomarker of specific fatty acid intake in middle-aged Japanese men. AB - OBJECTIVE: To assess the utility of serum phospholipid fatty acid (FA) levels as a biochemical indicator of habitual dietary fatty acid intake in Japanese, whose diet is characterized by low fat intake and high intake of n-3 polyunsaturated fatty acids (PUFA) of marine origin. SUBJECTS AND METHODS: Eighty-seven male volunteers from four public health center districts that were part of the Japan Public Health Center based Prospective Study (JPHC Study) cohort I, were included in this study. Habitual intake of fatty acid was obtained by 7 day weighed dietary records four times (in one area only twice) in 1994--1995. Blood was collected twice, in February and August of the same year, and the composition of FA in serum phospholipid was analyzed by gas chromatography. The correlation coefficient between serum phospholipid FA levels and fatty acid intake was calculated. RESULTS: High correlations were observed for eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA), which are marine origin n-3 PUFA (r=0.75, 0.49, 0.50, respectively). No significant correlation was observed for saturated fatty acid (SFA), although the monounsaturated fatty acid (MUFA), palmitoleic acid and oleic acid intake were moderately correlated (r=0.22, 0.35, respectively). The correlations for EPA, DPA and DHA were similar in both samples collected in February and August. CONCLUSIONS: These data suggest that in populations with a high and stable over time intake of n-3 PUFA of marine origin, a single measurement of serum phospholipids reflects the ranking of habitual intake of marine origin n-3 PUFA. PMID- 11477463 TI - Expression of ferritin receptor in placental microvilli membrane in pregnant women with different iron status at mid-term gestation. AB - OBJECTIVE: The effect of iron status in pregnant women on expression of ferritin receptor in placental microvilli membrane at mid-term gestation was investigated. DESIGN: Ferritin receptor binding sites and dissociation constants (K(d)) were determined in specimens of placental microvilli from 30 pregnant women at mid term gestation and six women at term-delivery. RESULTS: The ferritin receptor binding sites in the placental microvilli membrane in pregnant women with mild iron deficiency and moderate iron deficiency anemia were significantly higher then those in pregnant women with normal iron status. However, no significant difference was found between pregnant women with severe iron deficiency anemia and with normal measurements. No significant differences of K(d) values were detected between pregnant women with normal iron status and those with iron deficiency. Data also revealed that the ferritin receptor binding sites in placental microvilli membrane and K(d) values at mid-term gestation did not differ significantly from those at term gestation. CONCLUSION: Lower iron status in pregnant women could lead to an increase in expression of ferritin receptor in placental microvilli membrane at mid-term gestation while the dissociation constant of ferritin receptor remains unchanged. This implies that the regulation of maternal-fetal iron homeostasis via the ferritin receptor-mediated pathway is achieved by changes in the numbers of ferritin receptors rather then binding properties. PMID- 11477464 TI - The effect of a biscuit with red palm oil as a source of beta-carotene on the vitamin A status of primary school children: a comparison with beta-carotene from a synthetic source in a randomised controlled trial. AB - OBJECTIVE: To determine the effect of a biscuit with red palm oil as a source of beta-carotene on the vitamin A status of primary school children and to compare this with the effect of a biscuit with beta-carotene from a synthetic source. DESIGN: Randomised controlled trial. SETTING: A rural community in KwaZulu-Natal, South Africa. SUBJECTS: A total of 437 primary school children, aged 5--11 y; 400 completed the study. INTERVENTION: Subjects were randomly assigned to one of three groups, receiving, respectively: (i) a placebo biscuit; (ii) a biscuit with synthetic beta-carotene as a vitamin A fortificant (SB); (iii) a biscuit with red palm oil as a source of beta-carotene (PB); SB and PB supplied 30% of the RDA for vitamin A per serving of three biscuits. Biscuits were distributed daily during the school week; vitamin A status was assessed at baseline and after 3 months. RESULTS: There was a significant improvement in serum retinol compared to the control group in both the SB and PB groups (P<0.005); the treatment effect for the two groups was equivalent (difference in treatment effect 0.62 microg/dl, with a 90% CI -0.33--1.57). CONCLUSIONS: A biscuit with red palm oil as a source of beta-carotene is as effective as a biscuit with synthetic beta-carotene in improving the vitamin A status of primary school children. The additional qualities of red palm oil (ie no trans fatty acids; rich source of antioxidants) make it an excellent alternative fortificant for addressing vitamin A deficiency. PMID- 11477465 TI - Age-related differences in fat-free mass, skeletal muscle, body cell mass and fat mass between 18 and 94 years. AB - OBJECTIVE: To determine (1) lean and fat body compartments, reflected by fat-free mass (FFM), appendicular skeletal muscle mass (ASMM), body cell mass (BCM), total body potassium (TBK), fat mass and percentage fat mass, and their differences between age groups in healthy, physically active subjects from 18 to 94 y of age; and (2) if the rate of decrease in any one of the parameters by age might be accelerated compared to others. METHODS: A total of 433 healthy ambulatory Caucasians (253 men and 180 women) aged 18--94 y were measured by dual-energy X ray absorptiometry (DXA) and whole body scintillation counter (TBK counter) using a large sodium iodide crystal (203 mm diameter). RESULTS: The ASMM change (-16.4 and -12.3% in men and women, respectively) in >75 y-old compared to 18 to 34-y old subjects was greater than the FFM change (-11.8 and -9.7% in men and women, respectively) and this suggests that skeletal muscle mass decrease in older subjects was proportionally greater than non-skeletal muscle mass. BCM (-25.1 and -23.2% in men and women, respectively) and TBK differences were greater than the differences in FFM or ASMM suggesting altered composition of FFM in older subjects. Women had lower peak FFM, ASMM, BCM and TBK than men. CONCLUSIONS: The decline in FFM, ASMM, BCM and TBK is accelerated in men and women after 60 y of age and FFM, ASMM, BCM and TBK are significantly lower than in younger subjects. Fat mass continued to increase until around 75 y. PMID- 11477466 TI - Basal metabolic rate in children with a solid tumour. AB - OBJECTIVE: To study the level of and changes in basal metabolic rate (BMR) in children with a solid tumour at diagnosis and during treatment in order to provide a more accurate estimate of energy requirements for nutritional support. DESIGN: An observational study. SETTING: Tertiary care at the Centre for Paediatric Oncology, University Hospital Nijmegen. SUBJECTS: Thirteen patients were recruited from a population of patients visiting the University Hospital Nijmegen for treatment. All patients asked to participate took part in and completed the study. INTERVENTION: BMR was measured by indirect calorimetry, under stringent, standardised conditions, for 20 min and on three different occasions in all patients. Continuous breath gas analysis using a mouthpiece was performed. Weight, height and skinfold measurements were performed before each measurement. MAIN OUTCOME MEASURES: BMR was expressed as percentage of the estimated reference value, according to the Schofield formulas based on age, weight and sex, and in kJ (kcal) per kg of fat-free mass. RESULTS: At diagnosis, the BMR was higher than the estimated reference BMR in all patients and 44% of the patients were considered hypermetabolic. Mean BMR (as percentage of reference) was significantly increased (11.6% (s.d. 6.7%); P=0.001), but decreased during treatment in 12 of the 13 patients (mean decrease 12.7% (s.d. 3.9%); P<0.0001). Furthermore, a significant negative correlation (P=-0.67; P=0.01) was found between the change in BMR and tumour response. CONCLUSIONS: These data suggest that the BMR of children with a solid tumour is increased at diagnosis and possibly during the first phase of oncologic treatment. This may be important when determining energy requirements for nutritional support. PMID- 11477467 TI - An analysis of intra-uterine growth retardation in rural Malawi. AB - OBJECTIVE: (1) To describe the sex-specific, birth weight distribution by gestational age of babies born in a malaria endemic, rural area with high maternal HIV prevalence; (2) to assess the contribution of maternal health, nutritional status and obstetric history on intra-uterine growth retardation (IUGR) and prematurity. METHODS: Information was collected on all women attending antenatal services in two hospitals in Chikwawa District, Malawi, and at delivery if at the hospital facilities. Newborns were weighed and gestational age was assessed through post-natal examination (modified Ballard). Sex-specific growth curves were calculated using the LMS method and compared with international reference curves. RESULTS: A total of 1423 live-born singleton babies were enrolled; 14.9% had a birth weight <2500 g, 17.3% were premature (<37 weeks) and 20.3% had IUGR. A fall-off in Malawian growth percentile values occurred between 34 and 37 weeks gestation. Significantly associated with increased IUGR risk were primiparity relative risk (RR) 1.9; 95% CI 1.4--2.6), short maternal stature (RR 1.6; 95% CI 1.0--2.4), anaemia (Hb<8 g/dl) at first antenatal visit (RR 1.6; 95% CI 1.2--2.2) and malaria at delivery (RR 1.4; 95% CI 1.0--1.9). Prematurity risk was associated with primiparity (RR 1.7; 95% CI 1.3--2.4), number of antenatal visits (RR 2.2; 95% CI 1.6--2.9) and arm circumference <23 cm (RR 1.9; 95% CI 1.4 -2.5). HIV infection was not associated with IUGR or prematurity. CONCLUSION: The birth-weight-for-gestational-age, sex-specific growth curves should facilitate improved growth monitoring of newborns in African areas where low birth weight and IUGR are common. The prevention of IUGR requires improved malaria control, possibly until late in pregnancy, and reduction of anaemia. PMID- 11477468 TI - Energy cost of physical activity in cystic fibrosis. AB - OBJECTIVE: The purpose of this study was to compare the energy cost of standardized physical activity (ECA) between patients with cystic fibrosis (CF) and healthy control subjects. DESIGN: Cross-sectional study using patients with CF and volunteers from the community. SETTING: University laboratory. SUBJECTS: Fifteen patients (age 24.6+/-4.6 y) recruited with consent from their treating physician and 16 healthy control subjects (age 25.3+/-3.2) recruited via local advertisement. INTERVENTIONS: Patients and controls walked on a computerised treadmill at 1.5 km/h for 60 min followed by a 60 min recovery period and, on a second occasion, cycled at 0.5 kp (kilopond), 30 rpm followed by a 60 min recovery. The ECA was measured via indirect calorimetry. Resting energy expenditure (REE), nutritional status, pulmonary function and genotype were determined. RESULTS: The REE in patients was significantly greater than the REE measured in controls (P=0.03) and was not related to the severity of lung disease or genotype. There was a significant difference between groups when comparing the ECA for walking kg radical FFM (P=0.001) and cycling kg radical FFM (P=0.04). The ECA for each activity was adjusted (ECA(adj)) for the contribution of REE (ECA kJ kg radical FFM 120 min(-1)--REE kJ kg radical FFM 120 min(-1)). ECA(adj) revealed a significant difference between groups for the walking protocol (P=0.001) but no difference for the cycling protocol (P=0.45). This finding may be related to the fact that the work rate during walking was more highly regulated than during cycling. CONCLUSIONS: ECA in CF is increased and is likely to be explained by an additional energy-requiring component related to the exercise itself and not an increased REE. PMID- 11477469 TI - Food and nutrient intake in a cohort of 8-month-old infants in the south-west of England in 1993. AB - OBJECTIVE: To investigate food and nutrient intakes in 8-month-old infants. DESIGN: Prospective study. SETTING: Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC), south-west England. SUBJECTS: A total of 1131 singleton Caucasian infants (82% of those invited) from a 10% random sample of ALSPAC, known as Children in Focus (CIF). METHODS: Diet was assessed using a structured 3 day unweighed dietary record. Food and nutrient intakes were compared with intakes from the 6--9 month age group of a British infant feeding survey, which formed part of the National Diet and Nutrition Survey (NDNS). Nutrient intakes were compared with dietary reference values (DRV). RESULTS: Intakes of energy and most nutrients were very similar between CIF and NDNS. The main difference was in the type of fat eaten resulting in a higher polyunsaturated to saturated fatty acid ratio in CIF (0.34) compared with NDNS (0.21). Other differences included the much lower calcium and iodine intakes in CIF compared with the NDNS. Differences in the proportion of consumers of formula and cow's milk accounted for most of the nutrient differences. Energy intakes were similar to the estimated average requirements (EAR), however, breastfed infants were slightly below and non-breastfed were slightly above the EAR. Mean intakes of zinc and vitamin D were below the Reference Nutrient Intakes. CONCLUSIONS: The diets of 8 month-old infants in this study were adequate in most nutrients. Breastfed infants had slightly lower energy intakes than non-breastfed infants. PMID- 11477470 TI - Energy costs of standard activities among Indian adults. AB - OBJECTIVES: To estimate the energy cost of resting (RMR), sitting and standing for urban Indian adults and compare these estimates with the reported values. DESIGN: Energy costs were measured using oxylog while body fat was estimated using equipment (HBF300, OMRON Corporation, Japan) that works on the principle of bioelectrical impedance, for 24 men and 40 women, aged 20-50 y, engaged in sedentary activities. SETTINGS: Agharkar Research Institute, Pune, India. RESULTS: Mean energy cost (kJ/min) of resting (RMR), sitting and standing were significantly (P<0.01, for all) higher for men (4.01+/-0.42, 5.0+/-0.72 and 5.74+/-0.69, respectively) than women (3.54+/-0.28, 4.03+/-0.41 and 4.35+/-0.52, respectively). Gender difference increased with the level of activity, from 13% for RMR to 32% for standing. These differences reduced when adjusted (using analysis of covariance) for body weight and became non-significant on adjusting for fat-free mass (FFM) in the case of RMR and sitting activity. The measured values of energy cost (absolute and per kg weight) for these activities were similar to African subjects but lower compared to Asian or European subjects for both sexes. The stepwise regression analysis done separately by sexes showed weight (29%) in men and body mass index (44%) in women to be the best predictors of RMR, while regression analysis for combined sexes indicated FFM and height as predictors of RMR (r(2)=56%, P<0.01). If means to estimate body fat were not available, RMR could best be predicted with BMI and sex as predictors (r(2)=55%; P<0.01). This was mainly due to the fact that the sex differences in our population were more prominent in FFM than that in BMI. Our observations thus indicate the need to develop prediction equations separately for different populations owing to differences in their body compositions, especially in fat mass (FM) or FFM. CONCLUSION: The energy costs of activities were associated with body composition, especially with absolute fat-free mass, which may vary even with the same body fat percentage. Therefore, there is a need to develop separate prediction equations for different communities. PMID- 11477471 TI - Seasonality of growth in Shanghai infants (n=4128) born in 11 consecutive years. AB - OBJECTIVE: To describe the seasonal growth patterns in Shanghai infants, to explore seasonal time lag between weight gain and length gain, and to investigate the long-term effect of birth season on early postnatal growth. DESIGN: Community based longitudinal study. SETTING: Shanghai, People's Republic of China. METHOD: Children were followed up monthly from 1 to 6 months, 3 monthly from 6 to 12 months, and 6 monthly from 12 to 24 months. SUBJECTS: A total of 6018 children born between 1 January 1980 and 31 December 1990. MAIN OUTCOME MEASURES: Weight gain, length gain and change in body mass index (BMI) over the seasons of the year. RESULTS: The infants tended to grow faster in height in spring and summer, and faster in weight and BMI in autumn and winter. The seasonal effect on weight gain and length gain is largely independent. The mean length value at 1 month of age was about 2.0 cm higher in infants born in May to July than in those born in November to February. At 24 months of age this difference was reduced to about 0.7 cm. CONCLUSIONS: There is a clear and consistent seasonality in growth in Shanghai infants. The seasonality seems to act independently on weight and length. Birth month has some association with attained size, but this is reduced during the first 2 y of life. PMID- 11477472 TI - Structural characterisation of the distal 5' flanking region of the human interleukin-10 gene. AB - Interleukin-10 (IL-10) is an important immunoregulatory cytokine. The recent characterisation of the proximal 5' flanking region of IL-10 led to the identification of the promoter region. Two polymorphic dinucleotide repeats and 10 single nucleotide polymorphisms (SNPs) have been identified and suggested to be useful genetic markers in several diseases. We have sequenced a further 5275 bp from -9296 to -4021 of the distal part of the 5' flanking region of the human IL-10 gene from the cosmid clone pWE15-4/11. Our sequence analysis reveals a high density of Alu-repeats within the IL-10 gene locus, including three novel, related structures which we term Alu-IL10 (A-C). Using three overlapping PCR products spanning 5110 bp of this distal part of the IL-10 gene the following single base pair substitutions were identified: at -8571 C/T, -8531 G/A, -6752 A/T, -6208 G/C, -5402 C/G. In addition a heterozygous three base pair deletion at -7400 was observed. The SNPs at -8571 C/T and -8531 G/A are contained within an Alu-repeat. These data should further the understanding of how the IL-10 gene is controlled in man and how its function may vary between individuals. PMID- 11477473 TI - Genetic variation at the chemokine receptors CCR5/CCR2 in myocardial infarction. AB - Our objective was to examine the association between myocardial infarction (MI) and two DNA-polymorphisms at the proinflammatory chemokine receptors CCR2 (I64V) and CCR5 (32 bp deletion, (Delta)ccr5), defining if these polymorphisms influence the age for the onset of MI. A total of 214 patients with an age at the first MI episode <55 years, 96 patients that suffered the first MI episode when older than 60 years, and 360 population controls were polymerase chain reaction genotyped for the CCR2-V64I and CCR5-Delta32/wt polymorphisms. Patients and controls were male from the same Caucasian population (Asturias, northern Spain). The frequency of the Deltaccr5 allele was significantly higher in controls compared to patients <55 years (P = 0.004), or in patients >60 years compared to patients <55 years (P = 0.002). Taking the patients >60 years as the reference group, non-carriers of the (Delta)ccr5-allele would have a three-fold higher risk of suffering an episode of MI at <55 years of age (OR = 3.06; 95% CI = 1.46-6.42). Gene and genotype frequencies for the CCR2 polymorphism did not differ between patients <55 years and controls or patients >60 years. Our data suggest that the variation at the CCR5 gene could modulate the age at the onset of MI. Patients carrying the (Delta)ccr5-allele would be protected against an early episode of MI. CCR5 and the CCR5-ligands are expressed by cells in the arteriosclerotic plaque. Thus, the protective role of (Delta)ccr5 could be a consequence of an attenuated inflammatory response, that would determine a slower progression of the arteriosclerotic lesion among (Delta)ccr5-carriers. Our work suggests that the pharmacological blockade of CCR5 could be a valuable therapy in the treatment of MI. PMID- 11477474 TI - Association and linkage of leprosy phenotypes with HLA class II and tumour necrosis factor genes. AB - Previous analyses indicate major gene control of susceptibility to leprosy per se and the HLA class II region has been implicated in determining susceptibility and control of clinical phenotype. Segregation analysis using data from 76 Brazilian leprosy multi-case pedigrees (1166 individuals) supported a two locus model as the best fit: a recessive major gene and a recessive modifier gene(s) (single locus vs two locus model, P = 0.0007). Combined segregation and linkage analysis to the major locus, showed strong linkage to HLA class II (HLA-DQB1 P = 0.000002, HLA-DQA1 P = 0.000002, HLA-DRB1 P = 0.0000003) and tumour necrosis factor genes (TNF P = 0.00002, LTA P = 0.003). Extended transmission disequilibrium testing, using multiple affected family members, demonstrated that the common allele TNF*1 of the -308 promoter region polymorphism showed linkage and/or association with disease per se, at a high level of significance (P < 0.0001). Two locus transmission disequilibrium testing suggested susceptibility (TNF*1/LTA*2) and protective (TNF*2/LTA*2) haplotypes in the class iii region. Taken together the segregation and HLA analyses suggest the possibility of more than one susceptibility locus in the MHC. PMID- 11477475 TI - A genome screen for multiple sclerosis in Italian families. AB - We have screened the whole genome for linkage in 40 Italian multiplex families with multiple sclerosis using 322 markers. The GENEHUNTER-PLUS program was used to analyse these data and revealed eight regions of potential linkage where the lod score exceeds the nominal 5% significance level (0.7). No region of linkage with genome-wide significance was identified and none of the markers showed evidence of statistically significant transmission disequilibrium. Overall these results have refined the linkage data relating to this disease in Italians modestly supporting some previously identified areas of interest and helping to exclude others. PMID- 11477476 TI - Sequence analysis of the DRB1 promoter reveals limited polymorphism with no influence on gene expression. AB - HLA-class II promoters contain a set of conserved regulatory regions necessary for constitutive and induced gene expression. For the HLA-DQB as well as for the DRB1 promoter sequence, polymorphisms with influence on gene expression have been reported. In contrast to these data we could show that there is very limited allele-specific polymorphism among the HLA-DRB1 promoter alleles. In a long range PCR we amplified a DNA sequence containing the promoter and the second exon of the DRB1 gene in one fragment. Nested PCR products of this PCR fragment for the promoter and for the second exon were analysed by DNA sequencing to allow the linkage of a promoter to its DR allele. Most investigated DRB1 alleles exhibited the same promoter consensus sequence except for two point mutations. An A to T transversion (position -70 bp) was closely associated with DRB1*08, whereas a C deletion (position -30 bp) was most commonly observed together with DRB1*10. Both polymorphisms did not influence promoter activity in luciferase reporter gene assays. PMID- 11477477 TI - DQA1 and DQB1 promoter diversity and linkage disequilibrium with class II haplotypes in Mexican Mestizo population. AB - The upstream sequences in the 5' flanking region of HLA class II genes, regulate their expression and contribute to the development of immunological diseases. We analyzed 105 healthy unrelated Mexican Mestizos for QAP and QBP polymorphism. DNA typing for DRB1, DQA1, DQB1, QAP1 and QBP1 was done using a standardized PCR SSOP. Although all QAP alleles previously described were found in Mexicans, the distribution differed as compared to other populations. QAP-3.1, 4.1 and 4.2 were the most frequent alleles and were associated with DQA1*03, *0501 and *0402 respectively. The prevalent QBP alleles were 3.21, 3.1 and 4.1 found mainly associated with DQB1*0302, *0301 and *0501. Linkage disequilibria between the promoter and the corresponding DQA1 and DQB1 allele, are in general the same as described by others. A total of 61 different haplotypes were defined, only six of them with a frequency above 4%. The haplotypes DRB1*0407-QAP-3.1-DQA1*03-QBP-3.21 DQB1*0302 (HF = 14.37%) and DRB1*0802-QAP-4.2-DQA1*0401-QBP-4.1-DQB1*0402 (HF = 14.22%), which have an Amerindian ancestry, are the most frequent in Mexicans. Some rare combinations were detected such as DRB1*0405-QAP-1.3-DQA1*0101/4-QBP 5.11/5.12-DQB1*0501 and DRB1*0403-QAP-3.2-DQA1*03-QBP-3.21-DQB1*0302, probably due to ancient recombination events. This knowledge is relevant as a basis to evaluate functional implications and to explore the role of promoter diversity in disease expression. PMID- 11477478 TI - IL-1B and IL-1Ra gene polymorphisms and disease severity in rheumatoid arthritis: interaction with their plasma levels. AB - The balance between interleukin-1 (IL-1) and its competitive antagonist IL-1 receptor antagonist (IL-1Ra) may contribute to the pathogenesis of rheumatoid arthritis (RA). We analysed the frequency of different alleles in the IL-1B gene (at -511 and at +3954) as well as in the IL-1Ra gene (at +2018) in an association study involving 297 RA patients and 112 healthy controls from the same geographic area. We tested associations with RA susceptibility or severity, and with circulating levels of IL-1Ra and IL-1beta. Carriage of the rare IL-1B (+3954) allele 2 was increased in destructive arthritis (DRA) as compared to non destructive arthritis (NDRA) (OR 1.7, 95% CI 1.1-2.8, 49.0% vs 35.9%) and controls (OR 1.7, 95% CI 1.1-2.8, 35.8%). Patients carrying this allele had a more destructive (Larsen wrist radiological index: mean +/- s.e.m., 2.1 +/- 0.2 vs 1.6 +/- 0.1, P = 0.005; Steinbrocker functional index: 2.4 +/- 0.1 vs 1.9 +/- 0.1, P = 0.002) and active disease (Ritchie articular index: 8.1 +/- 0.8 vs 5.3 +/- 0.6, P = 0.002; erythrocyte sedimentation rate (ESR): 36.6 +/- 2.9 mm/h vs 25.3 +/- 1.8 mm/h, P = 0.002). This contribution was independent from that of HLA DR4/DR1 to severity. IL-1Ra plasma levels adjusted to ESR values were significantly lower in IL-1B2 (+3954) positive than negative RA patients (1.0 +/- 0.1 vs 1.2 +/- 0.1 ng/ml, P = 0.01). This IL-1B (+3954) gene polymorphism may be an important marker for the severity of joint destruction in RA and is associated with an imbalance in IL-1Ra production. As this genetic association was independent and additive to the risk of HLA DR4/DR1 status, it could be a useful addition to HLA-DR4/1 as a genetic prognostic marker early in the course of the disease. PMID- 11477479 TI - Polymorphisms of the tumor necrosis factor alpha locus among autoimmune disease susceptible and resistant inbred rat strains. AB - Inbred rat strains manifest remarkable differences in susceptibility/severity to autoimmune disease. MHC alleles strongly influence the pathogenesis of autoimmune disease in rats, but the precise mechanism(s) remain inadequately defined. The TNFalpha gene is located in the class III region of the MHC. Polymorphisms, influencing either the structure or expression of the TNF protein, might contribute to differences in autoimmune disease susceptibility/severity. We therefore sequenced the Tnf locus using genomic DNA from ACI, BB(DR), BN, DA, F344, and LEW rats that vary in susceptibility/severity to autoimmune diseases. We found 42 polymorphisms among these six strains. Although none of these polymorphisms are predicted to change the amino acid sequence of the TNF protein, several reside in potential non-coding regulatory regions and may influence expression levels. These polymorphisms may serve as good candidates for analysis of TNF expression to elucidate the mechanism(s) by which the MHC regulates susceptibility and/or severity of autoimmune diseases. PMID- 11477480 TI - Possible association between CTLA4 DNA polymorphisms and early onset type 1 diabetes in a UK population. AB - Linkage and association has been reported between CTLA4 DNA markers and susceptibility to type 1 diabetes in some populations, but not others. We performed case-control and family-based association studies to assess if the CTLA4 A49G and intron 1 C/T polymorphisms were associated with development of early onset type 1 diabetes in the Northern Ireland population. The distribution of A49G and C/T alleles in cases (n = 144) was similar to those observed in controls (n = 307). In contrast, significant distortions in allele transmissions from informative parents to probands were observed for both the A49G (P = 0.02) and C/T (P = 0.01) polymorphisms employing 297 nuclear families. Our results suggest that the CTLA4 gene may play a minor role in the overall genetic predisposition to type 1 diabetes in this UK population. PMID- 11477481 TI - Glutathione S-transferase genotypes in systemic sclerosis and their association with clinical manifestations in early disease. AB - The glutathione S-transferases (GSTs) are a family of enzymes involved in limiting oxidative damage to tissues. Null alleles for one or more of the GST enzymes, especially GSTM1, reportedly occur more frequently in patients with Sjogren's syndrome and systemic lupus erythematosus who possess certain autoantibodies. Because systemic sclerosis (SSc) is a disease in which oxidative damage has been hypothesized to contribute both to immune dysfunction and tissue damage, we sought to determine if patients from a multi-ethnic cohort of SSc patients with early disease (< or =5 years) were more likely than ethnically matched normal controls to have null alleles for GSTM1 (M1) and/or GSTT1 (T1), and if the null allele status correlated with any major disease features. The data show that while M1 and T1 null genotypes were not significantly increased in SSc compared to ethnically matched controls, their frequencies (especially T1 nulls) were significantly higher among SSc patients with hypertension and pulmonary involvement. This suggests that GST genotype may be a genetic factor that contributes to clinical disease expression in SSc. PMID- 11477485 TI - Apomorphine SL (Uprima): a new treatment for the management of erectile dysfunction. PMID- 11477486 TI - The history of erectile dysfunction management. AB - This article describes the evolution or revolution in the management of erectile dysfunction over the centuries. In recent history there has been a rapid movement away from a predominant role for the specialist towards the primary care physician. The physician is increasingly faced with the need to individualize therapy to meet patient expectations. PMID- 11477487 TI - Neurophysiology/pharmacology of erection. AB - Despite considerable advances, both the central regulation of erection with processing of various stimuli, and the different steps involved in neurotransmission, impulse propagation and intracellular transduction of neural signals in penile smooth muscles, are still incompletely known. Centrally as well as peripherally, many transmitters and transmitter systems are involved. Dopamine, nitric oxide, oxytocin and ACTH/alpha-MSH, seem to have a facilitatory role, whereas serotonin may be either facilitatory or inhibitory, and enkephalins are inhibitory. Peripherally, the balance between contractant (eg noradrenaline, endothelins, angiotensins) and relaxant (eg NO, VIP and related peptides, prostanoids) factors controls the degree of contraction of the smooth muscle of the corpora cavernosa, and determines the functional state of the penis. Neurogenic NO is considered the most important factor for relaxation of penile vessels and corpus cavernosum. The roles of other putative transmitters/mediators and of various intracellular mechanisms, producing relaxation of vascular and corpus cavernosum smooth muscle, have not been established. For example, recent findings have suggested a role of Rho/Rho-kinase in the regulation of cavernosal tone, and that Rho-kinase antagonism could be a new potential principle for the treatment of erectile dysfunction. Further research in this area may be rewarding. PMID- 11477488 TI - Dopamine and sexual function. AB - The use of the D1/D2 dopamine receptor agonist apomorphine SL for the treatment of erectile dysfunction provides a strong support in favour of a participation of the dopaminergic system in the control of sexual function. However, the exact involvement of dopamine in sexual motivation and in the control of genital arousal in humans is unknown. In contrast, experimental data suggest an implication of dopamine at all these stages of the copulatory behaviour in rodents. The release of dopamine at the level of the nucleus accumbens, which is innervated by the mesolimbic dopaminergic pathway originating in the ventral tegmental area, is positively implicated in the pre-copulatory or appetitive phase in male rats. There is also a permissive role in the copulatory or consumatory phase for dopamine released at the level of the median pre-optic area, which receives projection from the dopaminergic incertohypothalamic pathway within the hypothalamus. It is noteworthy that these participations of the dopaminergic system are not specific to sexual behaviour but rather reflect the more general involvement of dopamine in the regulation of cognitive, integrative and reward processes. Due to its role in the control of locomotor activity, the integrity of the nigrostriatal dopaminergic pathway is also essential for the display of copulatory behaviour. Somehow more specific to sexual function, it is likely that dopamine can trigger penile erection by acting on oxytocinergic neurons located in the paraventricular nucleus of the hypothalamus, and perhaps on the pro-erectile sacral parasympathetic nucleus within the spinal cord. The counterpart of such regulation of the genital arousal by dopamine has not yet been established in females. In conclusion, the central dopaminergic system is a key element of the control of sexual function. PMID- 11477489 TI - Apomorphine to Uprima: the development of a practical erectogenic drug: a personal perspective. AB - The development process for apomorphine SL as an effective treatment for patients with erectile dysfunction has been somewhat unusual. As often is the case, much of the impetus for the basic research originated in academia. However, somewhat unusually, the impetus for early stage clinical research also lay in the hands of the academics. This article represents a historical perspective from one of those involved throughout. PMID- 11477490 TI - Characterising the benefit of apomorphine SL (Uprima) as an optimised treatment for representative populations with erectile dysfunction. AB - The clinical profile for apomorphine sublingual (SL), a new centrally active agent for the management of the erectile dysfunction (ED) patient, is described in this article. Apomorphine SL is shown to be rapid in onset (71% of patients within 20 min) with a consistent, predictable response that is independent of severity (mild, moderate or severe), the underlying aetiology or the presence of significant co-morbidities (coronary artery disease, hypertension, etc). Importantly, there is also consistent long-term clinical benefit (>90% of attempts being successful over 18 months), for patients who respond to therapy and a benign side effect profile (<13.4% patients with adverse events). This formulation of apomorphine has a speed of onset and overall clinical profile that may offer particular advantages to the patient in terms of spontaneity and predictability of response. ED is a complex disease of varying aetiologies and severities often associated with a number of co-morbidities that require diverse solutions. Given the need for customisation of therapy to individual patient needs, the clinical profile of apomorphine SL would indicate that it will make a most welcome addition to the physician's armamentarium against ED. PMID- 11477491 TI - Safety and tolerability of apomorphine SL (Uprima). AB - The side effect profile of apomorphine SL (2-6 mg) has been determined in clinical studies of over 5000 patients using over 120 000 doses. Apomorphine, 2 and 3 mg, has been shown to have an excellent safety profile. The most commonly occurring side effects (<7%), nausea, headache and dizziness, tend to be mild and not compliance limiting. Neither the incidence nor the nature of the side effects is significantly affected by common co-morbidites or by the use of many concurrent medications. Over this dose range there is little evidence of vasoactivity; there is little change in haemodynamic baseline and there is no synergistic effect with nitrates. Although syncope can occur at higher doses, it is rarely observed at approved doses (<0.2%). PMID- 11477492 TI - The management of erectile dysfunction in the community. AB - Erectile dysfunction (ED) is a widely occurring benign disorder that affects men of all ages. The prevalence and severity of ED increases with age and results in considerable distress and impact on quality of life for those who suffer from it. As ED is associated with a wide variety of underlying conditions and co morbidities there is a requirement for diversity of treatment options beyond those currently available. In the management of the ED patient both primary care and specialist physicians have an important role to play. This article reports on a stepwise approach for the diagnosis and treatment of ED, with an emphasis on a 'shared care' approach. The suitability of apomorphine SL (Uprima) for the front line management of the ED patient is described. PMID- 11477493 TI - Low-dose growth hormone treatment combined with diet restriction decreases insulin resistance by reducing visceral fat and increasing muscle mass in obese type 2 diabetic patients. AB - OBJECTIVE: To evaluate the effects of low-dose growth hormone (GH) therapy combined with diet restriction on changes in body composition and the consequent change in insulin resistance in newly-diagnosed obese type 2 diabetic patients. DESIGN: Double-blind and placebo-controlled trial of 25-kcal/kg IBW diet daily with GH (n=9; rhGH, 0.15 IU/kg body weight/week) or placebo (n=9) for 12 weeks. SUBJECTS: Eighteen newly-diagnosed obese type 2 diabetic patients (age 42--56 y, body mass index 28.1+/-2.7 kg/m(2)). MEASUREMENTS: Body composition and fat distribution parameters (by bioelectrical impedance analyzer and CT scans), serum IGF-1; serum glucose, insulin and free fatty acid (FFA) during oral glucose tolerance test (OGTT); HbA(1c); serum lipid profiles; and glucose disposal rate (GDR) by euglycemic hyperinsulinemic clamp at baseline and after treatment. RESULTS: The fraction of body weight lost as fat lost was significantly greater (0.98+/-0.39 vs 0.52+/-0.32 kg/kg, P<0.05) and visceral fat area was decreased more in the GH-treated group compared to the placebo-treated group (27.9 vs 21.6%, P<0.05). Lean body mass and muscle area were reduced in the placebo treated group, whereas an increase in both was observed in the GH-treated group. GDR the was significantly increased in only the GH-treated group (4.67+/-1.05 vs 6.95+/-0.91 mg/kg/min, P<0.05). The GH-induced increase in GDR was positively correlated with the decrease in the ratio of visceral fat area/muscle area (r=0.588, P=0.001). Serum glucose levels and insulin- and FFA-area under the curve during OGTT and HbA(1c) were significantly decreased after GH treatment. LDL-cholesterol level was decreased in only the GH-treated group. CONCLUSION: Low dose GH treatment combined with dietary restriction resulted not only in a decrease of visceral fat but also in an increase of muscle mass with a consequent improvement of the insulin resistance observed in obese type 2 diabetic patients. PMID- 11477494 TI - Exercise training-induced triglyceride lowering negatively correlates with DHEA levels in men with type 2 diabetes. AB - OBJECTIVE: To investigate the effect of an exercise training program on lipid profile in correlation with DHEA level and body weight and body composition in type 2 diabetic men. DESIGN: Longitudinal, controlled clinical intervention study with exercise training consisting of an 8 week supervised program of aerobic exercise (75% VO(2) peak, 45 min), twice a week and intermittent exercise, once a week, on a bicycle ergometer. SUBJECTS: Sixteen men (age 45.4+/-7.2 y (mean+/ s.d.), HbA1c 8.15+/-1.7%, body mass index (BMI) 29.6+/-4.6 kg/m(2)) were randomly divided into two groups: trained group (n=8) and control group (n=8). MEASUREMENTS: Lipid, apo- and lipoprotein and DHEA concentrations. Cross sectional areas of subcutaneous and visceral adipose tissue and mid-thigh muscle by magnetic resonance imaging. RESULTS: Training decreased visceral (153.25+/ 38.55 vs 84.20+/-21.30 cm(2), P<0.001), subcutaneous (241.55+/-49.55 vs 198.00+/ 39.99 cm(2), P<0.001) adipose tissue area and triglyceride levels (2.59+/-1.90 vs 1.79+/-1.08 nmol/l, P<0.05) and increased mid-thigh muscle cross-sectional area (148.30+/-36.10 vs 184.35+/-35.85 cm(2), P<0.001), and DHEA levels (11.00+/-3.10 vs 14.25+/-4.10 nmol/l, P<0.05) with no modification in body weight. Changes in triglycerides were negatively correlated with changes in DHEA (r=-0.81, P=0.03). This correlation was independent of changes in abdominal fat distribution. CONCLUSION: Training decreases abdominal fat depots, improves muscular mass and affects favourably triglyceride and DHEA levels. Changes in triglycerides and DHEA were inversely related. PMID- 11477495 TI - Relationship of a large weight loss to long-term weight change among young and middle-aged US women. AB - OBJECTIVE: To assess the prevalence of clinically significant weight loss among women and whether this is associated with smaller long-term weight gains. DESIGN: Six-year follow-up of young and middle-aged women in the Nurses' Health Study II. SUBJECTS: A total of 47,515 women who did not report a pregnancy, or a diagnosis of cancer or cardiovascular disease any time between 1989 and 1995. MEASUREMENTS: Self-reported weights in 1989, 1991, 1993 and 1995, dietary intake, physical activity, inactivity, history of weight cycling and smoking. RESULTS: Between 1989 and 1991, 9% of the women lost > or =5% of their 1989 weight (6% lost 5- 9.9% and 3% lost > or =10%). The proportion who lost > or =10% of their weight increased with category of body mass index (BMI, kg/m(2)) from 0.4% among women with a BMI <22 to 9% among women with a BMI > or =30 in 1989. Women who lost > or =5% of their weight between 1989 and 1991 gained more weight between 1991 and 1995 than their peers and the difference increased across categories of BMI in 1989. However, due to their large weight losses, women who lost > or =5% of their weight between 1989 and 1991 overall gained less weight than their peers between 1989 and 1995 (P<0.001). Moreover, women who engaged in 5 or more hours per week of vigorous physical activity gained approximately 0.5 kg less than their inactive peers (P<0.001). CONCLUSION: Although most women who lost a clinically significant amount of weight regained most of it, they gained less weight over the entire 6 y period than their peers. PMID- 11477496 TI - Weight loss in overweight subjects following low-sucrose or sucrose-containing diets. AB - OBJECTIVES: To compare the response by overweight individuals, counselled in a work environment, to energy-reduced diets in which the amount of sucrose containing foods is varied. DESIGN: Two energy-reduced diets were designed as a weight-reducing programme. A low-sugar diet (LSD) providing 5% of its energy from sucrose and a sugar-containing diet (SCD) providing 10% of its energy from sucrose incorporated as sweet foods were devised. Both diets were constructed to contain about 33% of the energy from fat. The diets, designed to provide a deficit of 2.51 MJ/day (600 kcal/day) per individual, were randomly allocated to subjects in an 8 week parallel design study. SUBJECTS: Ninety-five subjects were recruited from a large service industry if they were more than 7 kg (1 stone) in weight above body mass index (BMI) 25 kg/m(2). Sixty-eight subjects completed the programme. MEASUREMENTS: Fortnightly body weight measurements were taken using calibrated scales; BMI at baseline and week 8; and nutrient intake using 2 day food record diaries at baseline and weeks 2, 4 and 8. RESULTS: Weight loss over the 8 weeks was 2.2 kg (LSD) and 3.0 kg (SCD). BMI changed from 29.2 on the LSD and 30.1 kg/m(2) SCD at baseline to 28.2 and 28.8 kg/m(2) at week 8 respectively. The actual prescribed commercially added sucrose intakes were 5% energy (LSD) or 10% energy (SCD). Reported percentage energy from fat was significantly lower on the SCD (and would seem to support the theory of an inverse relationship between fat and sugar) than on the LSD, where there was seen to be no significant reduction. There was no evidence of micronutrient dilution that could be directly attributed to the sucrose content of the diets. CONCLUSION: These results provide no justification for the exclusion of added sucrose in weight-reducing diets. PMID- 11477497 TI - Conjugated linoleic acid (CLA) reduced abdominal adipose tissue in obese middle aged men with signs of the metabolic syndrome: a randomised controlled trial. AB - BACKGROUND: Abdominal obesity is strongly related to metabolic disorders. Recent research suggests that dietary conjugated linoleic acid (CLA) reduces body fat and may improve metabolic variables in animals. The metabolic effects of CLA in abdominally obese humans have not yet been tested. OBJECTIVE: To investigate the short-term effect of CLA on abdominal fat and cardiovascular risk factors in middle-aged men with metabolic disorders. METHODS: Twenty-five abdominally obese men (waist-to-hip ratio (WHR), 1.05+/-0.05; body mass index (BMI), 32+/-2.7 kg/m(2) (mean+/-s.d.)) who were between 39 and 64-y-old participated in a double blind randomised controlled trial for 4 weeks. Fourteen men received 4.2 g CLA/day and 10 men received a placebo. The main endpoints were differences between the two groups in sagittal abdominal diameter (SAD), serum cholesterol, low-density lipoprotein, high-density lipoprotein, triglycerides, free fatty acids, glucose and insulin. RESULTS: At baseline, there were no significant differences between groups in anthropometric or metabolic variables. After 4 weeks there was a significant decrease in SAD (cm) in the CLA group compared to placebo (P=0.04, 95% CI; -1.12, -0.02). Other measurements of anthropometry or metabolism showed no significant differences between the groups. CONCLUSIONS: These results indicate that CLA supplementation for 4 weeks in obese men with the metabolic syndrome may decrease abdominal fat, without concomitant effects on overall obesity or other cardiovascular risk factors. Because of the limited sample size, the effects of CLA in abdominal obesity need to be further investigated in larger trials with longer duration. PMID- 11477498 TI - Effect of a six month gemfibrozil treatment and dietary recommendations on the metabolic risk profile of visceral obese men. AB - OBJECTIVE: The aim of the present study was to investigate the effects of a 6 month gemfibrozil treatment. SUBJECTS: A sample of 64 visceral obese men (age 46+/-6 y; body mass index 31+/-3 kg/m(2); waist circumference 104+/-7 cm; mean+/ s.d.) who received dietary recommendations. METHODS: Subjects were randomly assigned to receive a placebo (n=32) or gemfibrozil (1200 mg/day) (n=32). RESULTS: In both placebo and gemfibrozil treated groups, significant reductions were noted in body weight, fat mass, waist circumference and visceral adipose tissue area measured by computed tomography (0.00010.007 kg/l. Precision of FMI obtained using pairs of procedures improves to <4.5% in adults and <5.5% in children. PMID- 11477501 TI - The effect of leptin treatment on the development of obesity in overfed suckling Wistar rats. AB - OBJECTIVE: To identify the role of hyperleptinaemia in mediating the effects of early postnatal overfeeding in a rat strain known to be prone to manipulations of the early environment which result in predispositions for obesity and associated metabolic and cardiovascular disturbance in later life. DESIGN: Wistar rats were reared in normal litters (NL, 10--12 pups) or small litters (SL, four pups) from postnatal day 3 and killed for determination of body composition and plasma leptin and insulin concentrations on day 7 or day 21 after having been treated with recombinant leptin (2 x 50 (pmol/g)/day) or saline from day 1. RESULTS: Rearing in SL doubled the body fat content and plasma leptin levels in comparison to NL pups by 21 days of age. Under leptin-treatment throughout suckling age, NL pups remained leptin responsive, ie the difference in body fat content was progressively reduced relative to the controls. Until 7 days of age, despite the body fat content of untreated SL pups being 2-fold higher and their plasma leptin level 7-fold higher than that of NL pups, leptin treatment caused the same percentage decreases in body fat in SL than in NL pups. But in contrast to NL pups, the SL pups became leptin resistant thereafter. Plasma insulin levels in 7 day-old leptin-treated SL pups were 3-fold higher than in untreated littermates and 5-fold higher than in the NL groups. CONCLUSION: Prophylactic leptin treatment does not prevent hyperinsulinaemia and excessive fat deposition in SL pups. On the other hand, selective hyperleptinaemia during suckling age does not trigger leptin resistance and obesity in NL pups. Rather than hyperleptinaemia per se, other factors associated with early postnatal overnutrition, for example, the concurrent hyperinsulinaemia, seem to play a pivotal role for the development of leptin-resistance and life-long obesity risk in SL rats. PMID- 11477502 TI - Maternal obesity and pregnancy outcome: a study of 287,213 pregnancies in London. AB - OBJECTIVE: To examine the maternal and foetal risks of adverse pregnancy outcome in relation to maternal obesity, expressed as body mass index (BMI, kg/m(2)) in a large unselected geographical population. DESIGN: Retrospective analysis of data from a validated maternity database system which includes all but one of the maternity units in the North West Thames Region. A comparison of pregnancy outcomes was made on the basis of maternal BMI at booking. SUBJECTS: A total of 287,213 completed singleton pregnancies were studied including 176,923 (61.6%) normal weight (BMI 20--24.9), 79 014 (27.5%) moderately obese (BMI 25--29.9) and 31,276 (10.9%) very obese (BMI> or =30) women. MEASUREMENTS: Ante-natal complications, intervention in labour, maternal morbidity and neonatal outcome were examined and data presented as raw frequencies and adjusted odds ratios with 99% confidence intervals following logistic regression analysis to account for confounding variables. RESULTS: Compared to women with normal BMI, the following outcomes were significantly more common in obese pregnant women (odds ratio (99% confidence interval) for BMI 25--30 and BMI> or =30 respectively): gestational diabetes mellitus (1.68 (1.53--1.84), 3.6 (3.25--3.98)); proteinuric pre eclampsia (1.44 (1.28--1.62), 2.14 (1.85--2.47)); induction of labour (2.14 (1.85 -2.47), 1.70 (1.64--1.76)); delivery by emergency caesarian section (1.30 (1.25- 1.34), 1.83 (1.74--1.93)); postpartum haemorrhage (1.16 (1.12--1.21), 1.39 (1.32- 1.46)); genital tract infection (1.24 (1.09--1.41), 1.30 (1.07--1.56)); urinary tract infection (1.17 (1.04-1.33), 1.39 (1.18--1.63)); wound infection (1.27 (1.09--1.48), 2.24 (1.91--2.64)); birthweight above the 90th centile (1.57 (1.50- 1.64), 2.36 (2.23--2.50)), and intrauterine death (1.10 (0.94--1.28), 1.40 (1.14- 1.71)). However, delivery before 32 weeks' gestation (0.73 (0.65--0.82), 0.81 (0.69--0.95)) and breastfeeding at discharge (0.86 (0.84--0.88), 0.58 (0.56- 0.60)) were significantly less likely in the overweight groups. In all cases, increasing maternal BMI was associated with increased magnitude of risk. CONCLUSION: Maternal obesity carries significant risks for the mother and foetus. The risk increases with the degree of obesity and persists after accounting for other confounding demographic factors. The basis of many of the complications is likely to be related to the altered metabolic state associated with morbid obesity. PMID- 11477503 TI - Waist circumference vs body mass index for prediction of disease risk in postmenopausal women. AB - OBJECTIVE: To test the sensitivity of waist circumference (central adiposity) as an index of disease risk in postmenopausal women. DESIGN: Retrospective analysis of postmenopausal women tested at Washington University School of Medicine. SUBJECTS: A total of 323 healthy postmenopausal (66+/-5 y; mean+/-s.d.) women not using any hormone replacement. MEASUREMENTS: Body composition, hyperinsulinemia (insulin area), triglycerides and HDL-cholesterol. RESULTS: Excess waist size had a stronger association with hyperinsulinemia and hypertriglyceridemia than body mass index (BMI; kg/m(2)) in otherwise healthy, postmenopausal women. After adjusting for BMI, a strong relation existed between waist circumference and insulin area, HDL-cholesterol and triglycerides (P<0.01). Conversely, after adjusting for waist circumference, no relation was apparent between BMI and the dependent variables of interest. The strength of the association between waist circumference and disease risk became most apparent when analyses were restricted to normal-weight women (BMI 24--28 kg/m(2)). When BMI was held constant, hyperinsulinemia and triglyceridemia increased dose-dependently with changes in waist size. CONCLUSION: Waist circumference, an easily obtained index of central adiposity, is a more sensitive measure of relative disease risk than is BMI in middle-aged and older women, particularly in normal-weight individuals. PMID- 11477504 TI - Prevalence and clustering of cardiovascular risk factors among healthy adults in a Chinese population: the MJ Health Screening Center Study in Taiwan. AB - OBJECTIVE: To gain insight into the prevalence and clustering of multiple cardiovascular risk factors in a healthy Chinese adult population in Taiwan. DESIGN: A cross-sectional study was carried out in 1996. SUBJECTS: A total of 46,603 subjects (23,485 men and 23,118 women) who were aged 20--59 y and attended a private health screening center for health examination of their own volition. MEASUREMENTS: Multiple cardiovascular risk factors including cigarette smoking, overweight (23 kg/m(2)< or =body mass index (BMI)<25 kg/m(2)) and obesity (BMI> or =25 kg/m(2)), lipid disorder (a ratio of total cholesterol level to the level of high density lipoprotein cholesterol>5 or use of lipid-lowering drugs), hypertension (systolic blood pressure> or =140 mmHg or diastolic blood pressure> or =90 mmHg or use of anti-hypertensive medications), and diabetes mellitus (fasting serum plasma glucose level> or =126 mg/dl or use of anti-diabetic medications) were determined. RESULTS: In comparison to women, men had a higher prevalence of current smoking (42.1 vs 5.6%), overweight (25.1 vs 17.1%) and obesity (33.1 vs 21.5%), lipid disorder (45.1 vs 19.6%), hypertension (17.4 vs 13.2%), as well as diabetes mellitus (4.1 vs 3.4%). The prevalence of men or women having two or more of the cardiovascular risk factors of interest was 54.3 and 21.7%, respectively. With advancing age, the prevalence of risk factors became greater for both genders. More importantly, the clustering of risk factors increased monotonically with increasing BMI levels for men and women. CONCLUSIONS: The prevalence and clustering of cardiovascular risk factors are commonplace in this healthy Chinese adult population. Considering the significant association between clustering of risk factors under study and BMI levels, this study gives an indication that population-based multifactorial interventions may work out favorably for specific groups. PMID- 11477505 TI - Weight-related differences in glucose metabolism and free fatty acid production in two South African population groups. AB - OBJECTIVE: The effects of free fatty acids (FFA), leptin, tumour necrosis factor (TNF) alpha and body fat distribution on in vivo oxidation of a glucose load were studied in two South African ethnic groups. DESIGN AND MEASUREMENTS: Anthropometric and various metabolic indices were measured at fasting and during a 7 h oral glucose tolerance test (OGTT). Body composition was measured using bioelectrical impedance analysis and subcutaneous and visceral fat mass was assessed using a five- and two-level CT-scan respectively. Glucose oxidation was evaluated by measuring the ratio of (13)CO(2) to (12)CO(2) in breath following ingestion of 1-(13)C-labelled glucose. SUBJECTS: Ten lean black women (LBW), ten obese black women (OBW), nine lean white women (LWW) and nine obese white women (OWW) were investigated after an overnight fast. RESULTS: Visceral fat levels were significantly higher (P<0.01) in obese white than black women, despite similar body mass indexes (BMIs). There were no ethnic differences in glucose oxidation however; in the lean subjects of both ethnic groups the area under the curve (AUC) was higher than in obese subjects (P<0.05 for both) and was found to correlate negatively with weight (r=-0.69, P<0.01) after correcting for age. Basal TNF alpha concentrations were similar in all groups. Percentage suppression of FFAs at 30 min of the OGTT was 24+/-12% in OWW and -38+/-23% (P<0.05) in OBW, ie the 30 min FFA level was higher than the fasting level in the latter group. AUC for FFAs during the late postprandial period (120--420 min) was significantly higher in OWW than OBW (P<0.01) and LWW (P<0.01) and correlated positively with visceral fat mass independent of age (r=0.78, P<0.05) in the OWW only. Leptin levels were higher (P<0.01) both at fasting and during the course of the OGTT in obese women from both ethnic groups compared to the lean women. CONCLUSIONS: Glucose oxidation is reduced in obese subjects of both ethnic groups; inter- and intra-ethnic differences were observed in visceral fat mass and FFA production and it is possible that such differences may play a role in the differing prevalences of obesity-related disorders that have been reported in these two populations. PMID- 11477506 TI - The role of postprandial releases of insulin and incretin hormones in meal induced satiety--effect of obesity and weight reduction. AB - BACKGROUND: Previous studies have indicated that the secretion of the intestinal satiety hormone glucagon-like peptide-1 (GLP-1) is attenuated in obese subjects. OBJECTIVE: To compare meal-induced response of GLP-1 and glucose-dependent insulinotropic polypeptide (GIP) in obese and lean male subjects, to investigate the effect of a major weight reduction in the obese subjects, and to look for an association between these hormones and ad libitum food intake. METHOD: Plasma concentrations of intestinal hormones and appetite sensations were measured prior to, and every 30 min for 180 min after, ingestion of a 2.5 MJ solid test meal. Gastric emptying was estimated scintigraphically. An ad libitum lunch was served 3 h after the test meal. SUBJECTS: Nineteen non-diabetic obese (body mass index (BMI) 34.1--43.8 kg/m(2)) and 12 lean (BMI 20.4--24.7 kg/m(2)) males. All obese subjects were re-examined after a mean stabilised weight loss of 18.8 kg (95% CI 14.4--23.2). RESULTS: Total area under the GLP-1 response curve (AUC(total, GLP 1)) was lower in obese before and after the weight loss compared to lean subjects (P<0.05), although weight loss improved the response from 80 to 88% of that of the lean subjects (P=0.003). The GIP response was similar in obese and lean subjects. However, after the weight loss both AUC(total, GIP) and AUC(incremental, GIP) were lowered (P<0.05). An inverse correlation was observed between AUC(incremental, GIP) and energy intake at the subsequent ad libitum meal in all groups. In lean subjects ad libitum energy intake was largely predicted by the insulin response to the preceding meal (r(2)=0.67, P=0.001). CONCLUSION: Our study confirmed previous findings of a reduced postprandial GLP-1 response in severely obese subjects. Following weight reduction, GLP-1 response in the obese subjects apparently rose to a level between that of obese and lean subjects. The data suggests that postprandial insulin and GIP responses are key players in short-term appetite regulation. PMID- 11477507 TI - Effects of test-meal palatability on compensatory eating following disguised fat and carbohydrate preloads. AB - OBJECTIVE: To test whether the palatability of a test meal altered compensatory eating following disguised high-energy fat and carbohydrate preloads. DESIGN: Effects of preload energy (low, 265 kJ, or high, 1510 kJ) and test-meal palatability (bland or palatable) were contrasted within-subjects, with a between subjects contrast of fat and carbohydrate preloads. SUBJECTS: Twenty-four healthy, normal men (age 23.6+/-1.0 y, (body mass index) BMI 21.3+/-0.5). MEASUREMENTS: Microstructural analysis of test meal intake and rated appetite in the four test conditions, plus diary-based weighed intake analysis of energy intake post-lunch. RESULTS: Subjects ate significantly less at lunch after disguised high-energy fat or carbohydrate preloads relative to the low-energy preload, and ate significantly more of the palatable than bland lunch in all conditions. The reduction in eating following the high-energy preload was significantly less in the palatable condition. Intake post-lunch did not differ between conditions, and overall subjects had higher daily energy intake on the days they consumed the high-energy preloads. Rated hunger was significantly less 30 min after the high- than low-energy preloads, but increased on tasting the palatable food in all conditions. The high-energy preloads suppressed appetite immediately post-lunch. No differences between fat and carbohydrate were found on any measure. CONCLUSIONS: Manipulation of the palatability of a test meal modified the ability to respond to disguised high-energy preloads, with over consumption most evident when disguised high-energy preloads were followed by a palatable food. Subsequent voluntary intake compensated for over-consumption of the palatable lunch, but not the high-energy preload. PMID- 11477508 TI - Systemic nicotine stimulates human adipose tissue lipolysis through local cholinergic and catecholaminergic receptors. AB - OBJECTIVE: To evaluate whether the lipolytic effects of systemic nicotine are not only attributed to indirect adrenergic mechanisms, but also to a direct action of nicotine on fat cells. DESIGN: The effect of a systemic nicotine infusion (0.5 microg/kg/min for 30 min) on lipolysis in subcutaneous adipose tissue was investigated in situ in 11 non-obese, non-smoking, healthy male subjects under placebo-controlled conditions. MEASUREMENTS: By using microdialysis probes the glycerol levels (lipolysis index) and blood flow were monitored locally in subcutaneous adipose tissue. RESULTS: Plasma nicotine levels peaked (7.2 ng/ml) at the end of the infusion. Nicotine induced a mean (+/-s.e.) percentage peak increase in adrenaline and noradrenaline plasma levels of 213+/-30% (P<0.01) and 118+/-5% (P<0.05), respectively. Nicotine increased venous plasma glycerol levels by 144+/-9% (P<0.001), arterialized plasma glycerol levels by 148+/-12% (P<0.001) and adipose glycerol levels by 148+/-16% (P<0.001), but did not alter blood flow. By inducing a local cholinoceptor blockade with mecamylamine (10(-5) M) via the microdialysis system, the increase in adipose glycerol levels was inhibited by approximately 45% (P=0.02). A corresponding local beta-adrenoceptor blockade with propranolol (10(-4) M), inhibited the increase in adipose glycerol levels by approximately 60% (P=0.02). Infusion of saline (ie placebo) had no effect on the parameters mentioned above. CONCLUSION: Systemically administered nicotine induces lipolysis, in part by activating the classical adrenergic mechanism (mediated by a nicotine-induced release of catecholamines stimulating beta adrenoceptors), and in part by directly activating a nicotinic cholinergic lipolytic receptor located in adipose tissue. PMID- 11477509 TI - Effects of 36 hour fasting on GH/IGF-I axis and metabolic parameters in patients with simple obesity. Comparison with normal subjects and hypopituitary patients with severe GH deficiency. AB - OBJECTIVE: Reduction of growth hormone (GH) secretion in obesity probably reflects neuroendocrine and metabolic abnormalities. Even short-term fasting stimulates GH secretion and distinguishes normal from hypopituitary subjects with growth hormone deficiency (GHD). Marked weight loss improves GH secretion in obesity but the effect of fasting is controversial. We studied the effects of a 36 h fasting on the GH/IGF-I axis and metabolic parameters in obesity. SUBJECTS: We studied nine obese patients (OB; three male and six female; age, 29.2+/-4.8; range, 18-59 y; body mass index (BMI), 43.4+/-2.7 kg/m(2); WHR, 0.9+/-0.1). Fifteen normal subjects (NS; eight male and seven female 28.9+/-0.6, 25-35 y; 21.6+/-0.4 kg/m(2)) and 10 adult hypopituitary patients with severe GH deficiency (GHD; seven male and three female; 37.6+/-2.3, 29-50 y; 24.5+/-1.0 kg/m(2); GH peak<3 microg/l after ITT and/or<9 microg/l after GHRH+arginine) served as control groups. STUDY DESIGN: We studied the effects of 36 h fasting on 8 h diurnal mean GH, insulin and glucose concentrations (mGHc, mINSc and mGLUc; assay every 30 min from 8.00 am to 4.00 pm) as well as on IGF-I, IGFBP-3, ALS, IGFBP-1, GHBP and free fatty acid (FFA) levels. RESULTS: Before fasting, basal IGF-I and ALS levels in OB were similar to those in NS and both were higher (P<0.001) than those in GHD. IGFBP-3 levels in OB were lower (P<0.01) than in NS but higher (P<0.02) than in GHD. GHBP levels in OB and GHD were similar and both were higher (P<0.01) than in NS. Glucose levels were similar in all groups. FFA levels in OB were higher (P<0.01) than in NS but similar to those in GHD. IGFBP-1 in OB were lower (P<0.05) than in NS and GHD which, in turn, were similar. On the other hand, mINSc in OB was higher (P<0.01) than that in NS and GHD which, in turn, were similar. The mGHc in OB was similar to that in NS but only the latter was higher (P<0.05) than in GHD. The individual mGHc in the three groups overlapped. After fasting, IGF-I levels in GHD were unchanged while they decreased in OB (P=NS) as well as in NS (P<0.01). IGFBP-3 and ALS levels did not change. GHBP levels in OB and GHD were unchanged while they increased in NS (P<0.01). Glucose and FFA levels were reduced and increased, respectively, in all groups (P<0.02 and P<0.01). IGFBP-1 increased while mINSc decreased in all groups (P<0.02 and P<0.01); in OB they persisted lower and higher (P<0.01) respectively, than in NS and GHD. Fasting significantly increased mGHc in NS (P<0.001) but not in OB as well as in GHD. Individual mGHc in OB showed persistent overlap with GHD. CONCLUSIONS: Short-term fasting does not increase GH secretion in obesity and does not distinguish somatotroph function in obese from that in severe GHD adults. Short-term fasting in obesity has attenuated effects on insulin and IGFBP 1 secretion while it normally increases free fatty acids in spite of any change in GH secretion. PMID- 11477510 TI - Validation of deuterium labeled fatty acids for the measurement of dietary fat oxidation: a method for measuring fat-oxidation in free-living subjects. AB - BACKGROUND: Fatty acid oxidation has frequently been measured using (13)C or (14)C labeled fatty acids. While providing valuable data, the breath test method is hindered by the need for a controlled environment to measure VCO(2) and collect frequent breath samples. Additionally, the CO(2) breath tests require the use of (13)C- or (14)C-acetate to correct for isotope exchange in the TCA cycle. We validated d(31)-palmitic acid for measuring dietary fat oxidation. When oxidized, the deuterium appears as water and mixes with the body water pool providing a cumulative record of fat oxidation. METHODS: The recovery of deuterium from d(31)-palmitic acid at 10 h post-dose was compared to that of (13)CO(2) from [1-(13)C]-palmitic acid in nine subjects (body mass index (BMI)=23.6+/-2.8; percentage body fat (%BF)=22.6+/-5.3; mean+/-s.d.). Subjects were studied at rest. [1-(13)C]-acetate (2 mg/kg) was dosed in a liquid breakfast. On a second day, [1-(13)C]-palmitic acid (10 mg/kg) and d(31)-palmitic acid (15 mg/kg) were dosed with the same liquid breakfast. RESULTS: Recovery of (13)CO(2) from [1-(13)C]-acetate at 10 h post-dose was 53.7+/-10.4%. Recovery of d(31)-palmitic acid was 13.2+/-7.7% (mean+/-s.d.) and [1-(13)C]-palmitic acid recovery was 6.4+/-3.6%. When the (13)C data was corrected for [1-(13)C]-acetate (Na salt) recovery, the mean difference in percentage recovery between the two tracers was 0.5+/-2.8% and cumulative recoveries through 10 h post-dose were highly correlated (y=1.045x - 0.47; r(2)=0.88, P<0.0002). Our data shows both labels to be equivalent in their ability to measure dietary fat oxidation in resting subjects. CONCLUSIONS: The use of deuterium labeled palmitic acid eliminates the need for rigid control over the subjects' environment. Frequent sampling and measurement of VCO(2) are not needed for accurate calculation of percentage recovery of the deuterium label. In addition, the deuterium label has a decreased potential for isotopic exchange compared to (13)C or (14)C, so a recovery correction factor is probably not required. PMID- 11477511 TI - Weighing the care: physicians' reactions to the size of a patient. AB - OBJECTIVE: To examine how the weight of a patient affects both the attitudes that physicians hold as well as the treatments that they intend to prescribe. DESIGN: In a six-cell randomized design, physicians evaluated a medical chart of a male or female patient, depicted as either average weight, overweight or obese, who presented with a migraine headache. SUBJECTS: A total of 122 physicians affiliated with one of three hospitals located in the Texas Medical Center of Houston completed the experiment. MEASUREMENTS: Using a standard medical procedure form, physicians indicated how long they would spend with the patient and which of 41 medical tests and procedures they would conduct. They also indicated their affective and behavioral reactions to the patient. RESULTS: The weight of a patient significantly affected how physicians viewed and treated them. Although physicians prescribed more tests for heavier patients, F(2, 107)=3.65, P<0.03, they simultaneously indicated that they would spend less time with them, F(2, 107)=8.38, P<0.001, and viewed them significantly more negatively on 12 of the 13 indices. CONCLUSION: This study reveals that physicians continue to play an influential role in lowering the quality of healthcare that overweight and obese patients receive. As the girth of America continues to increase, continued research and improvements in the quality of such healthcare deserve attention. PMID- 11477512 TI - Health professionals' views of overweight people and smokers. AB - OBJECTIVES: To examine health professionals' views of overweight people, to compare these to their views of smokers, and to explore the role of level of severity on these perceptions. DESIGN: A postal survey of health professionals employing a two by two, independent factorial design. The health category (overweight or smoker) was divided by level of severity (moderate or extreme), so that respondents received questionnaires about either: (i) moderately overweight people; (ii) extremely overweight people; (iii) moderate smokers; or (iv) heavy smokers. PARTICIPANTS: Two-hundred and fifty-five general medical practitioners and clinical psychologists in the north of England. MEASUREMENTS: A questionnaire was designed to explore beliefs about the causes, attitudes towards, and perceptions of responsibility of overweight people and smokers. RESULTS: Moderately and extremely overweight people were perceived as having reduced self esteem, sexual attractiveness and health, and to be moderately responsible for changing their situation (but less so than smokers). There were clear level effects in the perceptions of overweight, but not so for smokers. Of the four groups, moderately overweight people were viewed most positively and extremely overweight (obese) people were viewed least positively. CONCLUSIONS: Overall, health professionals' attitudes to overweight people were neutral to negative rather than entirely negative. However, where apparent negative attitudes were more likely to be directed at obese people than moderately overweight people. As obesity is a risk to health, the practice implications of health professionals' negative attitudes or patients' reticence to visit professionals who treat them with disregard must be addressed. PMID- 11477513 TI - Effects of socioeconomic status on the obesity knowledge of adolescents from six Latin American cities. AB - OBJECTIVE: To examine the effects of socioeconomic status (SES) on the obesity knowledge of adolescents in six Latin American cities. DESIGN: Data were collected using an anonymous, self-administered questionnaire consisting of demographic questions and a 25-item multiple-choice obesity knowledge test. Test items were clustered under five topics: the fat and calorie content of foods and beverages; weight loss methods; energy expenditure; food preparation methods; and the relationship between obesity and health. SUBJECTS: A total of 1272 ninth grade students from higher and lower SES families were recruited at schools in Buenos Aires, Argentina (n=195); Guatemala City, Guatemala (n=212); Havana, Cuba (n=213); Lima, Peru (n=218); Panama City, Panama (n=195); and Santiago, Chile (n=239). RESULTS: Mean test scores reflected a low level of obesity knowledge among adolescents from higher and lower SES groups in all six cities. Nevertheless, a trend for higher scores emerged in favor of adolescents from wealthier families. This income effect persisted after controlling for gender and weight status. The weakest knowledge areas among youth from the higher SES groups were food preparation methods and the relationship between obesity and health while those for adolescents from the lower SES groups were the fat and calorie content of foods and beverages and the relationship between obesity and health. Classroom instruction about obesity was generally more available to students from the higher SES groups. The majority of adolescents from both SES groups were interested in learning more about weight loss methods, energy expenditure, and the fat and calorie content of foods and beverages. The topic of least interest was the relationship between obesity and health. CONCLUSION: These preliminary findings suggest a need for more obesity education programs for adolescents, especially for those living in poverty. PMID- 11477514 TI - U.S. dietary exposures to heterocyclic amines. AB - Heterocyclic amines (HAs) formed in fried, broiled or grilled meats are potent mutagens that increase rates of colon, mammary, prostate and other cancers in bioassay rodents. Studies of how human dietary HA exposures may affect cancer risks have so far relied on fairly crudely defined HA-exposure categories. Recently, an integrated, quantitative approach to HA-exposure assessment (HAEA) was developed to estimate compound-specific intakes for particular individuals based on corresponding HA-concentration estimates that reflect their meat-type, intake-rate, cooking-method and meat-doneness preferences. This method was applied in the present study to U.S. national Continuing Survey of Food Intakes by Individuals (CSFII) data on meats consumed and cooking methods used by >25,000 people, after adjusting for underreported energy intake and conditional on meat doneness preferences estimated from additional survey data. The U.S. population average lifetime time-weighted average of total HAs consumed was estimated to be approximately 9 ng/kg/day, with 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) estimated to comprise about two thirds of this intake. Pan-fried meats were the largest source of HA in the diet and chicken the largest source of HAs among different meat types. Estimated total HA intakes by male vs. female children were generally similar, with those by (0- to 15-year-old) children approximately 25% greater than those by (16+-year-old) adults. Race-, age- and sex-specific mean HA intakes were estimated to be greatest for African American males, who were estimated to consume approximately 2- and approximately 3-fold more PhIP than white males at ages <16 and 30+ years, respectively, after considering a relatively greater preference for more well-done items among African Americans based on national survey data. This difference in PhIP intakes may at least partly explain why prostate cancer (PC) kills approximately 2-fold more African American than white men, in view of experimental data indicating that PhIP mutates prostate DNA and causes prostate tumors in rats. PMID- 11477515 TI - Personal exposures to PM(2.5) and polycyclic aromatic hydrocarbons and their relationship to environmental tobacco smoke at two locations in Greece. AB - In the context of a large-scale molecular epidemiology study of biomarkers of genotoxicity of air pollution, 24-h mean personal exposures to airborne PM(2.5) (particulate matter <2.5 microm) and associated polycyclic aromatic hydrocarbon (PAHs) were measured in 194 non-smoking technical institute students living in the city of Athens, Greece (an area with moderately high levels of air pollution) and the nearby small town of Halkida anticipated to have lower pollution levels. Extensive information relevant to the assessment of long-term and recent exposure to PAH was obtained from questionnaires as well as a time-location-activity diary (TLAD) which was kept by all subjects during a 4-day observation period. During the last 24 h of this period, subjects underwent personal exposure monitoring for PM(2.5) and PAH, while a sample of blood was donated at the end of this period. All subjects were monitored in this way twice; once during a winter season (October-February) and once during the following summer season (June-September). Nine subjects with plasma cotinine levels above 20 ng/ml were considered as unreported smokers and excluded from the study. Winter PM(2.5) exposures were lower in Athens (geometric mean 39.7 microg/m(3)) than Halkida (geometric mean 56.2 microg/m(3)) (P<0.001), while there was no significant location difference during the summer (Athens: geometric mean 32.3 microg/m(3), Halkida: geometric mean 32.9 microg/m(3); P=0.79). On the other hand, PAH exposures (sum of the eight carcinogenic PAHs) were significantly higher in Athens than in Halkida during the winter (Athens: geometric mean 8.26 ng/m(3), Halkida: geometric mean 5.80 ng/m(3); P<0.001) as well as during the summer (Athens: geometric mean 4.44 ng/m(3), Halkida: geometric mean 1.48 ng/m(3); P<0.001). There was a significant difference in the profile of the PAH exposures at the two locations, the proportion of lighter PAH (benzo[a]anthracene, chrysene [CHRYS], benzo[k]fluoranthene, and benzo[b]fluoranthene) being higher, and that of heavier PAH (benzo[ghi]perylene [BPer] and indeno[1,2,3,cd]pyrene) lower, in Halkida than in Athens, regardless of season. This difference appeared to be related to individual exposure to environmental tobacco smoke (ETS), as indicated by (a) the correlation at the individual level between the CHRYS/BPer ratio and declared time of recent exposure to ETS as well as plasma cotinine levels, especially during the winter; (b) the parallel variation of the mean levels of all three markers (declared ETS exposure, cotinine levels, CHRYS/BPer ratio) among three subgroups of subjects (Athens subjects who had lowest levels of all three markers; Halkida subjects other than those living in the institute campus area; and Halkida subjects living in the institute campus area who had the highest levels of all three markers). This demonstrates that ETS can have a distinctive effect on the PAH exposure profile of subjects exposed to relatively low levels of urban air pollution. PMID- 11477516 TI - Fingerprinting PCB patterns among Mohawk women. AB - This study examined the association of contaminated fish consumption and polychlorinated biphenyl (PCB) body burden by comparing the similarity of the congener pattern in yellow perch, caught near the point source of industrial pollution, and in other local fish to the pattern found in the breast milk of Mohawk women from Akwesasne, a Native American community located along the St. Lawrence River in New York, Ontario, and Quebec. The similarity is defined by the weighted Euclidean distance between two congener patterns. Ninety-seven Mohawk mothers participated and provided samples of breast milk. One hundred fifty-four nursing women from the Supplemental Nutrition Program for Women, Infants and Children (WIC) of Warren and Schoharie counties, New York, who gave birth during the same time period, were used as the comparison group. Results revealed that the breast milk of the Mohawk women, who ate the most local fish, had a congener pattern that more closely resembled that of perch caught near the waste site or average sampled fish caught in the Reserve than Mohawk women who ate less fish or the controls. The outcome demonstrates how PCBs may be "fingerprinted" as they migrate offsite from industrial sources and ultimately result in human exposure. PMID- 11477517 TI - Exposure analysis of five fish-consuming populations for overexposure to methylmercury. AB - Mercury, in the form of methylmercury, is found in a myriad of fish species consumed by recreational and subsistence fishers around the world. Many agencies have attempted to address the issue of mercury consumption, resulting at times in the placement of advisories on waterways used for fishing. In this study, consumption rates of three Native American populations and two recreational fishing populations consuming freshwater or saltwater fish species were examined. These consumption rates were combined with fish contamination data to assess the level of exposure to methylmercury and to determine if any of these populations exceed a derived tolerable daily intake (TDI) for methylmercury (0.035 to 0.08 microg/kg/day). The TDI is unlikely to result in adverse health effects and is based on scientific studies investigating sensitive endpoints in children of mothers who consume fish over prolonged periods of time. Results from the exposure analysis indicate that many within the Native American populations exceed the TDI. This occurs even though the mercury concentrations in certain fish species are comparable to concentrations found in fish from open waters where "background" levels are assumed. Recreational anglers consuming freshwater species have exposure levels below the TDI as do nearly all anglers consuming saltwater species. Similar populations or populations having comparable consumption patterns of fish with equal or higher mercury levels in other areas will also exceed the TDI level for mercury. The public health implications of this exposure analysis are discussed. PMID- 11477518 TI - Use of global positioning system technology to track subject's location during environmental exposure sampling. AB - Global positioning system (GPS) data recorders were worn by subjects in the Oklahoma Urban Air Toxics Study (OUATS) for automatic logging of their location as they went about their normal daily activities. The location information obtained by the GPS units had an uncertainty of about 10-20 m, which was sufficiently precise to track subjects' movements on trips outside the immediate vicinity of their homes. Due to instrument problems, primarily related to reduced battery life, the units operated for only about 30% of the total monitoring time attempted in 25 trials. The GPS data were compared to time-activity diaries kept by the subjects. In almost all cases, the GPS data confirmed all travel events reported in the subjects' diaries. Additionally, in five out of five trials in which the logging period covered most or all of the subjects' daytime activities, at least one travel event that was not recorded in the diary was detected by GPS. Notwithstanding the limitations of present technology, GPS was found to be a promising means for tracking of research subjects in community-based exposure assessment studies. PMID- 11477519 TI - Exposures to nitrogen dioxide in EXPOLIS-Helsinki: microenvironment, behavioral and sociodemographic factors. AB - Personal exposures to nitrogen dioxide (NO(2)) were monitored for 176 randomly selected inhabitants (25-55 years old) of Helsinki Metropolitan area as a part of the EXPOLIS (Air Pollution Exposure Distributions Within Adult Urban Populations in Europe) study between October 1996 and December 1997. NO(2) measurements were 48-h averages collected by Palmes passive sampler tubes. Differences in personal exposures to NO(2) were analyzed between sub-populations stratified by microenvironment, behavioral, socioeconomic and demographic factors. Factors significantly associated with differences in exposures to NO(2) were home and work location, housing characteristics, traffic volume near home, season and keeping windows open at home. Exposure to environmental tobacco smoke (ETS) and use of gas stove were also associated with increased personal exposures, although only few participants had a gas stove in Helsinki, and other gas appliances are non-existent. Single adults had higher average exposures to NO(2) than married or cohabiting participants, suggesting differences in living conditions between these two groups. Increased education was associated with decreased exposures to NO(2) and employed men were more exposed than unemployed men. Increased exposures to NO(2) were not associated with age or occupational status in Helsinki. Thus, behavioral and sociodemographic factors may have significant impact on personal exposures to NO(2) and should be considered in addition to environmental determinants in any monitoring program. PMID- 11477520 TI - Exposure to tap water during pregnancy. AB - Studies of disinfection byproducts (DBPs) in drinking water and risk of adverse reproductive outcome have usually relied on approximate measures of exposure. Individual differences in consumption of bottled or filtered water, variability in tap water consumption at home and at work, dermal and inhalation exposure to volatile contaminants, and changes in residency during pregnancy may lead to exposure misclassification. We characterized exposures to tap water and other risk factors among 71 pregnant and 43 non-pregnant women attending public health clinics. Nearly all residences had a municipal water source, but 25% of women drank filtered or bottled water. Fifty percent of the women in our sample reported working outside the home where, on average, one third of their daily water intake took place. Pregnant women consumed more water than non-pregnant women (3.4 vs. 3.0 total l/day), especially cold tap water at home (1.8 vs. 1.3 l/day, 95% CI for the difference=0.1, 0.9). Patterns of showering were similar for both groups of women, but pregnant women were more likely to bathe and to bathe more frequently. The prevalence of smoking was lower among pregnant women (22.5% vs. 32.6%), as was the consumption of alcohol (4.2% vs. 53.5%, 95% CI for the difference=-64.9, -33.7). Thirty-two percent of women had moved during their current pregnancy. The data reaffirm the importance of collecting individual level data for water consumption and exposure to potential confounders to avoid misclassification bias. This study is the first to target women of low socio economic status (SES) and therefore of particular interest in studies of adverse reproductive outcomes for which this group is at increased risk. PMID- 11477521 TI - The National Human Activity Pattern Survey (NHAPS): a resource for assessing exposure to environmental pollutants. AB - Because human activities impact the timing, location, and degree of pollutant exposure, they play a key role in explaining exposure variation. This fact has motivated the collection of activity pattern data for their specific use in exposure assessments. The largest of these recent efforts is the National Human Activity Pattern Survey (NHAPS), a 2-year probability-based telephone survey (n=9386) of exposure-related human activities in the United States (U.S.) sponsored by the U.S. Environmental Protection Agency (EPA). The primary purpose of NHAPS was to provide comprehensive and current exposure information over broad geographical and temporal scales, particularly for use in probabilistic population exposure models. NHAPS was conducted on a virtually daily basis from late September 1992 through September 1994 by the University of Maryland's Survey Research Center using a computer-assisted telephone interview instrument (CATI) to collect 24-h retrospective diaries and answers to a number of personal and exposure-related questions from each respondent. The resulting diary records contain beginning and ending times for each distinct combination of location and activity occurring on the diary day (i.e., each microenvironment). Between 340 and 1713 respondents of all ages were interviewed in each of the 10 EPA regions across the 48 contiguous states. Interviews were completed in 63% of the households contacted. NHAPS respondents reported spending an average of 87% of their time in enclosed buildings and about 6% of their time in enclosed vehicles. These proportions are fairly constant across the various regions of the U.S. and Canada and for the California population between the late 1980s, when the California Air Resources Board (CARB) sponsored a state-wide activity pattern study, and the mid-1990s, when NHAPS was conducted. However, the number of people exposed to environmental tobacco smoke (ETS) in California seems to have decreased over the same time period, where exposure is determined by the reported time spent with a smoker. In both California and the entire nation, the most time spent exposed to ETS was reported to take place in residential locations. PMID- 11477522 TI - Modeled estimates of chlorpyrifos exposure and dose for the Minnesota and Arizona NHEXAS populations. AB - This paper presents a probabilistic, multimedia, multipathway exposure model and assessment for chlorpyrifos developed as part of the National Human Exposure Assessment Survey (NHEXAS). The model was constructed using available information prior to completion of the NHEXAS study. It simulates the distribution of daily aggregate and pathway-specific chlorpyrifos absorbed dose in the general population of the State of Arizona (AZ) and in children aged 3-12 years residing in Minneapolis-St. Paul, Minnesota (MSP). Pathways included were inhalation of indoor and outdoor air, dietary ingestion, non-dietary ingestion of dust and soil, and dermal contact with dust and soil. Probability distributions for model input parameters were derived from the available literature, and input values were chosen to represent chlorpyrifos concentrations and demographics in AZ and MSP to the extent possible. When the NHEXAS AZ and MSP data become available, they can be compared to the distributions derived in this and other prototype modeling assessments to test the adequacy of this pre-NHEXAS model assessment. Although pathway-specific absorbed dose estimates differed between AZ and MSP due to differences in model inputs between simulated adults and children, the aggregate model results and general findings for simulated AZ and MSP populations were similar. The major route of chlorpyrifos intake was food ingestion, followed by indoor air inhalation. Two-stage Monte Carlo simulation was used to derive estimates of both inter-individual variability and uncertainty in the estimated distributions. The variability in the model results reflects the difference in activity patterns, exposure factors, and concentrations contacted by individuals during their daily activities. Based on the coefficient of variation, indoor air inhalation and dust ingestion were most variable relative to the mean, primarily because of variability in concentrations due to use or no-use of pesticides. Uncertainty analyses indicated a factor of 10-30 for uncertainty of model predictions of 10th, 50th, and 90th percentiles. The greatest source of uncertainty in the model stems from the definition of no household pesticide use as no use in the past year. Because chlorpyrifos persists in the residential environment for longer than a year, the modeled estimates are likely to be low. More information on pesticide usage and environmental concentrations measured at different post-application times is needed to refine and evaluate this and other pesticide exposure models. PMID- 11477523 TI - Complications of meningococcal disease in college students. AB - A retrospective study was conducted to provide a description of the risk, complications, fatality, and sequelae associated with invasive meningococcal disease in college students admitted in the Allegheny county (Pennsylvania) hospital system from January 1990 to May 1999. PMID- 11477524 TI - Pneumococcal bacteremia in adults in Soweto, South Africa, during the course of a decade. AB - We retrospectively reviewed 414 episodes of pneumococcal bacteremia that occurred in adults from July 1986 through June 1987 (1986/1987) and from July 1996 through June 1997 (1996/1997) to monitor the incidence and clinical and laboratory characteristics and to assess the influence of human immunodeficiency virus (HIV) infection on any changes. The incidence increased from 26 per 100,000 persons in 1986/1987 to 36 per 100,000 persons in 1996/1997; the increase was most marked among patients who were aged 25-44 years (24 cases per 100,000 persons to 45 per cases 100,000 persons) and > or =65 years (43 cases per 100,000 persons to 50 cases per 100,000 persons). Of 161 patients who were tested for HIV in 1996/1997, 108 (67%) were HIV seropositive. Among the general population, the prevalence of other underlying diseases and smoking decreased from 45% and 67%, respectively, in 1986/1987 to 23% (P<.0001) and 35% (P<.0001) in 1996/1997. Strains of pneumococci that were not susceptible to penicillin were found in 4% patients in 1986/1987 and 12% in 1996/1997 (P=.005). This increase occurred exclusively among the HIV-infected patients (22% of the HIV-seropositive patients versus 4% of HIV seronegative patients; P=.008), and there was a parallel increase for childhood serotypes (51% of HIV-seropositive patients versus 17% of HIV-seronegative patients; P<.0001). PMID- 11477525 TI - Phase II double-blind, placebo-controlled study of the safety and efficacy of cidofovir topical gel for the treatment of patients with human papillomavirus infection. AB - Genital condylomata acuminata are nonmalignant human papillomavirus (HPV)-induced tumors in which HPV types 6 and 11 are most commonly found. Usual treatments for condylomata acuminata are nonspecific and are based on the destruction or removal of infected tissue. These procedures are often painful and are characterized by a high relapse rate. We report here what is to our knowledge the first double blind, placebo-controlled study of the use of cidofovir, a nucleotide analogue, for the treatment of genital papillomavirus infections. Thirty patients were enrolled in the study; 19 received cidofovir, and 11 received placebo. The median number of warts and the median baseline wart area were comparable for both groups. Nine (47%) of 19 patients in the cidofovir group had a complete response (total healing), compared with 0 of the patients in the placebo group (P=.006). None of the patients in the cidofovir group experienced progression of the disease, compared with 5 (45%) of 11 patients in the placebo group. The side effects recorded for both groups were comparable. PMID- 11477526 TI - Cryptococcosis in human immunodeficiency virus-negative patients in the era of effective azole therapy. AB - We conducted a case study of human immunodeficiency virus (HIV)-negative patients with cryptococcosis at 15 United States medical centers from 1990 through 1996 to understand the demographics, therapeutic approach, and factors associated with poor prognosis in this population. Of 306 patients with cryptococcosis, there were 109 with pulmonary involvement, 157 with central nervous system (CNS) involvement, and 40 with involvement at other sites. Seventy-nine percent had a significant underlying condition. Patients with pulmonary disease were usually treated initially with fluconazole (63%); patients with CNS disease generally received amphotericin B (92%). Fluconazole was administered to approximately two thirds of patients with CNS disease for consolidation therapy. Therapy was successful for 74% of patients. Significant predictors of mortality in multivariate analysis included age > or =60 years, hematologic malignancy, and organ failure. Overall mortality was 30%, and mortality attributable to cryptococcosis was 12%. Cryptococcosis continues to be an important infection in HIV-negative patients and is associated with substantial overall and cause specific mortality. PMID- 11477527 TI - Occupational transmission of Mycobacterium tuberculosis to health care workers in a university hospital in Lima, Peru. AB - From November 1996 through March 1997, presumptive active pulmonary tuberculosis (TB) was detected in 44 health care workers (HCWs) at a university hospital in Lima, Peru. To further assess the magnitude of the outbreak and determine risk factors for occupational Mycobacterium tuberculosis transmission, we identified HCWs in whom active pulmonary TB was diagnosed from January 1994 through January 1998, calculated rates by year and hospital work area, and conducted a tuberculin skin test (TST) survey. Thirty-six HCWs had confirmed active pulmonary TB. The rate of TB was significantly higher among the 171 HCWs employed in the laboratory than among HCWs employed in all other areas. In multivariate analysis, the only independent risk factor for HCW M. tuberculosis infection in HWCs employed in the laboratory was the use of common staff areas. Very high rates of active pulmonary TB were detected among HCWs at this hospital, and occupational acquisition in the laboratory was associated with HCW-to-HCW transmission. PMID- 11477528 TI - Clinical significance of donor-unrecognized bacteremia in the outcome of solid organ transplant recipients. AB - We evaluated the clinical significance of unrecognized bacteremia in the organ donor (i.e., blood culture results that were reported to be positive after transplantation) on the outcome of transplant recipients. Twenty-nine of 569 liver and heart donors (5%) had bacteremia at the time of organ procurement, but there were no documented instances of transmission of the isolated bacteria from the donor to the recipient. Unrecognized bacteremia in the donor does not have a negative clinical impact on the outcome of organ transplant recipients. PMID- 11477529 TI - Prevalence of vaginal colonization by drug-resistant Candida species in college age women with previous exposure to over-the-counter azole antifungals. AB - We enrolled 382 college-age women in a cross-sectional survey to investigate the relationship between use of over-the-counter (OTC) azole-based antifungal drugs and vaginal colonization by drug-resistant Candida. This study showed no correlation (P=.506) between previous OTC exposure and colonization of drug resistant Candida in vaginal flora. However, a small number of resistant Candida species isolates were obtained from women with a history of multiple exposures to OTC antifungals; given the widespread use of these products, this may be an emerging concern. PMID- 11477530 TI - Clinical spectrum and transmission characteristics of infection with Norwalk-like virus: findings from a large community outbreak in Sweden. AB - A large foodborne outbreak caused by Norwalk-like virus (NLV) among children and staff at 30 day care centers provided an opportunity to study symptomatology and attack rates among patients in different age groups, as well as secondary transmission rates in centers and households. A retrospective cohort study of 775 subjects from 13 randomly chosen centers was performed. Diarrhea was more common in adults than in children (P=.001), whereas the reverse was noted with regard to vomiting (P=.003). The primary attack rate was 27% (142 of 524 subjects): 54% of adults versus 19% of children (P<.001). The mean incubation time for foodborne cases of infection was 34 hours. The secondary attack rate was 17%. Risk factors for spread into households were the primary case being a child (relative risk [RR], 3.8; 95% confidence interval [CI], 1.9-7.6) and vomiting (RR, 2.4; 95% CI, 1.0-5.5). The incubation time for person-to-person transmission was approximated by a mean serial interval of 52 hours. This is the first reported outbreak of NLV infection in which secondary transmission into households by individuals has been studied. PMID- 11477531 TI - Lack of seroconversion in a health care worker after polymerase chain reaction documented acute hepatitis C resulting from a needlestick injury. AB - We present a case of documented acute hepatitis C that occurred in a health care worker who sustained a needlestick injury while caring for an individual who was infected with both hepatitis C virus (HCV) and human immunodeficiency virus (HIV). According to the findings of third-generation serological assays performed during a follow-up of >1 year, the health care worker, who was treated with interferon-alpha (during weeks 2-6) and ribavirin (during weeks 5-9), did not develop antibodies against HCV, in spite of documentation of an HCV-specific T cell response. PMID- 11477532 TI - Acute flaccid paralysis syndrome associated with echovirus 19, managed with pleconaril and intravenous immunoglobulin. AB - We describe a 39-year-old woman who had undergone bilateral lung and renal transplantation and who was admitted to the hospital with acute onset of flaccid paralysis of the left leg due to echovirus 19 infection. The patient was treated with pleconaril and intravenous immunoglobulin, which correlated with clinical and laboratory evidence of improvement. PMID- 11477533 TI - Trichosporon asahii, a non-Candida yeast that caused fatal septic shock in a patient without cancer or neutropenia. AB - Trichosporon asahii (formerly Trichosporon beigelii) is an emerging fungal pathogen seen particularly in immunologically compromised patients. There are now approximately 100 reported cases of hematogenously disseminated infections with this life-threatening yeast, and no effective antifungal therapy is available. The present case is unusual because the patient did not have neutropenia or evidence of a malignancy. PMID- 11477534 TI - Fulminant leptospirosis in a patient with human immunodeficiency virus infection: case report and review of the literature. AB - We report a case of fulminant leptospirosis that was acquired in New York City by a patient with underlying infection with human immunodeficiency virus (HIV). Review of the literature on leptospirosis in HIV-infected persons showed that all patients were severely ill but responded well to treatment, which highlights the importance of recognizing this potentially life-threatening illness, especially in unusual settings. PMID- 11477535 TI - Septic shock due to babesiosis. AB - We present the case of a 69-year-old man with an unremarkable past medical history and an intact spleen who developed shock and renal failure due to babesiosis. Despite hemodynamic parameters showing severe distributive shock with hyperdynamic cardiac function, the patient recovered fully after treatment with quinine sulfate and clindamycin. PMID- 11477536 TI - Interferon-alpha drives T cell-mediated immunopathology in the intestine. AB - The ability of interferon (IFN)-alpha to induce autoimmunity and exacerbate Th1 diseases is well known. We have recently described enhanced expression of IFN alpha in the mucosa of patients with celiac disease (CD), a gluten-sensitive Th1 mediated enteropathy, characterized by villous atrophy and crypt cell hyperplasia. Previous studies from this laboratory have shown that T cell activation in explant cultures of human fetal gut can also result in villous atrophy and crypt cell hyperplasia. We have, therefore, examined changes that take place in explant cultures of human fetal gut after activation of T cells with anti-CD3 and/or IFN-alpha. We show that activation of T cells with anti-CD3 alone elicits a small IFN-gamma and TNF-alpha response with no tissue injury. Similarly, no changes are seen in explants cultured with IFN-alpha alone. However, addition of IFN-alpha with anti-CD3 results in enhanced Th1 response and crypt cell hyperplasia. This is associated with enhanced phosphorylation of STAT1, STAT3, and Fyn, a Src homology tyrosine kinase, which interacts with both TCR and IFN-alpha signal components. Together these data indicate that IFN-alpha can facilitate activation of Th1-reactive cells in the gut and drive immunopathology. PMID- 11477537 TI - Naive CD4(+) lymphocytes convert to anergic or memory-like cells in T cell deprived recipients. AB - Recent demonstrations that naive T cells proliferate after transfer to lymphopenic hosts have led to the theory that active homeostatic mechanisms fill the peripheral pool of naive T cells. To extend these data, we injected naive CD4(+) T cells from AND TCR transgenic mice (H-2(b/b) or H-2(k/k)) into CD3 epsilon-deficient mice, and studied the absolute number, phenotype and functional capacities of the transferred lymphocytes, from the first days to a few months after transfer. Proliferation of naive CD4(+) T cells did not fill the peripheral naive T cell pool. Injected naive T cells acquired a memory-like phenotype that was stable with time, despite the absence of foreign antigenic stimulation. Their functional capacities were modified, enhanced or abolished depending on the MHC haplotype. Thus, "homeostatic" proliferation of naive CD4(+) T cells in T cell deprived recipients does not regenerate the naive CD4(+) T cell pool. PMID- 11477538 TI - Gold is a T cell polyclonal activator in BN and LEW rats but favors IL-4 expression only in autoimmune prone BN rats. AB - Gold salts are beneficial in the treatment of rheumatoid arthritis but may induce immune-mediated disorders in predisposed patients. Gold salts induce Th2 dependent autoimmunity in Brown-Norway (BN) rats but not in Lewis (LEW) rats. The aim of this study was to define molecular targets of gold salts and to approach why LEW rats are resistant. Gold salts act on early steps of transduction in T cells from BN and LEW rats since they trigger tyrosine phosphorylation of numerous proteins including p56(lck) and a calcium signal which results in IL-4 and IFN-gamma expression by BN and LEW T cells. However, the IL-4 response was favored in BN spleen cells in vitro and in vivo. IFN-gamma, produced in part by CD8(+) cells, contributes to the resistance of LEW rats since gold salt-injected LEW rats receiving anti-CD8 or anti-IFN-gamma mAb displayed the parameters characteristics of gold salt-induced Th2 autoimmunity although to a lesser extent than in BN rats. Gold salts transduce a signal in BN and LEW spleen cells resulting in IL-4 and IFN-gamma gene transcription with a preferential IL-4 response in BN rats, a Th2-prone strain, while IFN-gamma contributes to the resistance of LEW rats. PMID- 11477539 TI - Sperm protein 17 (Sp17) in multiple myeloma: opportunity for myeloma-specific donor T cell infusion to enhance graft-versus-myeloma effect without increasing graft-versus-host disease risk. AB - We recently found that sperm protein 17 (Sp17), a spermatozoa-restricted protein, is aberrantly expressed on the tumor cells in patients with multiple myeloma (MM). It may therefore be possible to generate donor-derived Sp17-specific CTL for administration following allogeneic stem cell transplant to augment graft versus-myeloma (GVM) effect without inducing a global GVHD. To assess this approach, we have produced recombinant Sp17 protein and used Sp17 protein-pulsed dendritic cells to generate HLA class I-restricted Sp17-specific CTL from a previously unimmunized healthy donor. These CTL were able to lyse autologous Epstein-Barr virus-transformed lymphoblastoid cells in a Sp17-dependent manner. Target lysis was HLA-A1 and HLA-B27 restricted. Cytotoxicity could be blocked by antibodies against monomorphic HLA class I, HLA-A1 and HLA-B27 molecules but not HLA class II molecules. Most importantly, the CTL lysed HLA class I-matched Sp17 positive tumor cells, suggesting that Sp17 is processed and presented in association with the HLA class I molecules in Sp17-positive tumor cells in a concentration and configuration that could be recognized by recombinant protein primed CTL. Analysis by flow cytometry of the CTL indicated that they were predominantly CD8 in phenotype and they produced IFN-gamma and very little IL-4. Our results suggest the potential for the generation and administration of donor derived Sp17-specific CTL to augment GVM without inducing GVHD following allogeneic stem cell transplant for MM. PMID- 11477540 TI - Immunodomination results from functional differences between competing CTL. AB - The presence of dominant epitopes suppresses generation of CTL activity toward other non-dominant epitopes found on the same antigen-presenting cell (APC). This phenomenon, termed immunodomination, drastically restricts the diversity of the repertoire of CTL responses. Under various experimental conditions we assessed the in vivo expansion by tetramer staining and function by expression of O glycans and intracellular perforin of CTL specific for a dominant (B6(dom1)) and a non-dominant (HY) H2D(b)-restricted epitope. Immunodomination abrogated expansion rather than differentiation of HY-specific CTL. When immunodomination was precluded because HY was presented alone or because high numbers of antigen bearing APC were present, the numbers of HY-specific T cells detected after antigen priming were similar to those of B6(dom1)-specific T cells. The main difference between T cells that recognized B6(dom1) versus HY was functional rather than quantitative. The key feature of T cells specific for B6(dom1) is that they show striking up-regulation of molecules involved in CTL effector activity rather than accumulating to particularly high levels, as assessed by tetramer staining. These results support the emerging concept that following antigen priming, CTL populations of similar size can display important differences in effector function, and suggest that these functional differences are instrumental in shaping the repertoire of CTL responses. PMID- 11477541 TI - CD4(+)CD8(+)TCR(low) thymocytes express low levels of glucocorticoid receptors while being sensitive to glucocorticoid-induced apoptosis. AB - While signaling by either the TCR or glucocorticoid receptor (GR) can induce apoptosis in thymocytes, recent studies have shown that combining these signals results in survival of CD4(+)CD8(+) thymocytes. Although glucocorticoids (GC) in this way may directly affect T cell selection, no data are available addressing GR expression in thymocyte subsets and in individual cells within subsets. We studied GR expression by combining immunofluorescence cell surface staining for CD4, CD8 and TCR with intracellular staining of GR in four-color cytometry. Significant differences of GR expression were observed in various thymocyte subsets, although a homogeneous distribution of GR expression in individual thymocyte subsets emerged. The highest GR expression was found in CD4(-)CD8( )TCR(-) thymocytes, and decreased during development via the CD4(-)CD8(+)TCR(-) subpopulation into the CD4(+)CD8(+)TCR(low) subset. Interestingly, the latter population, although expressing less than half the GR density of CD4(-)CD8(-)TCR( ) cells, is the most sensitive subset to GC-induced apoptosis. Up-regulation of TCR expression by the CD4(+)CD8(+)TCR(low) subset to CD4(+)CD8(+)TCR(high) cells was accompanied by a parallel increase in GR expression. The latter finding and the presence of a homogeneous distribution of GR in each thymocyte subset provides an experimental basis for the concept that GR can antagonize TCR mediated signals at a constant rate relative to TCR expression. PMID- 11477542 TI - Superantigen overcomes resistance of IL-6-deficient mice towards MOG-induced EAE by a TNFR1 controlled pathway. AB - Experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein peptide 35-55 (MOG) leads to a chronic form of disease characterized by demyelination, inflammation and gliosis in the central nervous system (CNS). Recently IL-6 and LT alpha were found to be required for induction of the disease. The main features associated with EAE resistance of IL-6(-/-) and LT alpha(-/-) mice were reduced T cell proliferation and endothelial activation. As shown here treatment of MOG-immunized IL-6(-/-) mice with staphylococcal enterotoxin B (SEB)reversed their resistance to MOG-induced EAE. SEB failed to restore susceptibility to EAE in LT alpha(-/-) mice. The effect of SEB to induce EAE in IL-6(-/-) mice depends on TNF receptor type 1 (TNFR1) signaling because IL 6/TNF/LT alpha(-/-) and IL-6/TNFR1(-/-) are refractory to SEB. TNFR1 is involved in SEB induced trafficking of T cells into the CNS as evidenced by the failure to up-regulate VCAM-1 on CNS endothelium and lack of accumulation of V beta 8(+) T cells in the CNS of IL-6/TNFR1(-/-) mice upon immunization with MOG and treatment with SEB. The course of SEB triggered EAE in MOG immunized IL-6(-/-) mice was characterized by reduced severity and duration of clinical manifestations, which were associated with a significant drop of CNS infiltrating neutrophils and MIP-2 expression after peak disease. Taken collectively the effect of SEB to overcome EAE resistance points to a transient IL-6 independent but TNFR1 dependent proinflamatory pathway in EAE pathogenesis and suggests a crucial function for IL 6 in disease perpetuation. PMID- 11477543 TI - Lipopolysaccharide induction of indoleamine 2,3-dioxygenase is mediated dominantly by an IFN-gamma-independent mechanism. AB - Indoleamine 2,3-dioxygenase (IDO) is a rate-limiting enzyme in the L-tryptophan kynurenine pathway, which converts an essential amino acid, L-tryptophan, to N formylkynurenine. It has been speculated that IFN-gamma is a dominant IDO inducer in vivo. The present study used IFN-gamma or TNF-alpha gene-disrupted mice and IFN-gamma antibody-treated mice to demonstrate that lipopolysaccharide (LPS) induced systemic IDO is largely dependent on TNF-alpha rather than IFN-gamma. IFN gamma-independent IDO induction was also demonstrated in vitro with LPS stimulated monocytic THP-1 cells. These findings clearly indicate that there is an IFN-gamma-independent mechanism of IDO induction in addition to the IFN-gamma dependent mechanism. PMID- 11477544 TI - Somatic diversity of the immunoglobulin repertoire is controlled in an isotype specific manner. AB - We have studied two aspects of the IgE immune response. First, we have compared the kinetics of the IgE response to the T cell-dependent antigen ph-Ox coupled to ovalbumin with that of the IgG1 response and we have assessed the quality of the IgE response. Second, we have studied the generation of somatic diversity, understood as the combined effect of somatic mutation and the selection of D(iversity) and J(oining) elements, in germinal center B cells at the molecular level, using the germ-line sequence of the prototype anti-ph-Ox heavy chain variable element V(H)Ox1 as reference. We evaluated sequences derived from mu-, gamma 1- and epsilon-variable elements and showed that somatic diversification was different for all isotypes studied. We further compared the IgE responses of wild-type mice with those of mice expressing a truncated cytoplasmic IgE tail (IgE(KVK Delta tail)). IgE(KVK Delta tail) mice showed a more diverse sequence pattern. We corroborated previous results suggesting that short CDR3 regions are indicative for high-affinity antibodies by measuring relative affinities of phage expressed Fab fragments with prototype long and short CDR3 regions. Therefore, the composition of the antigen-receptor is responsible for the selection process and the expansion of antigen-specific cells, leading to an isotype-specific antibody repertoire. PMID- 11477545 TI - Transplant tolerance modifying antibody to CD200 receptor, but not CD200, alters cytokine production profile from stimulated macrophages. AB - Increased C57BL/6 allograft survival following donor-specific dendritic cell (DC) portal vein (pv) pre-transplant immunization of C3H mice is associated with increased expression of the molecule CD200 on DC, delivery of suppressive signals by CD200(r+) macrophages, and polarization in cytokine production towards type-2 cytokines. Infusion of anti-mouse CD200 monoclonal antibody abolishes these effects. We have used whole Ig, and F(ab')(2) fragments, of anti-CD200 and anti CD200(r) mAb to explore the relative signaling role of CD200(+) versus CD200(r+) cells in suppression of type-1 cytokine production in mixed leukocyte cultures (MLC), and enhanced graft survival in vivo. Simple neutralization of CD200 [even by F(ab')(2) antibody] reversed CD200-mediated suppression. However, only whole anti-CD200(r) antibody was effective in stimulating suppression from CD200(r+) cells. Suppression of cytokine induction following cross-linking of CD200(r+) cells in vitro was attenuated by anti-IL-6 mAb. Our data are consistent with the hypothesis that CD200(r) itself delivers the crucial intracellular signal leading to immunosuppression, a feature likely of importance in autoimmunity and transplantation. PMID- 11477546 TI - Modulation of proteinase-K resistant prion protein by prion peptide immunization. AB - Prion diseases are caused by conformational alterations in the prion protein (PrP). The immune system has been assumed to be non-responsive to the self-prion protein, therefore, PrP autoimmunity has not been investigated. Here, we immunized various strains of mice with PrP peptides, some selected to fit the MHC class II-peptide binding motif. We found that specific PrP peptides elicited strong immune responses in NOD, C57BL/6 and A/J mice. To test the functional effect of this immunization, we examined the expression of proteinase-K-resistant PrP by a scrapie-infected tumor transplanted to immunized syngeneic A/J mice. PrP peptide vaccination did not affect the growth of the infected tumor transplant, but significantly reduced the level of protease-resistant PrP. Our results demonstrate that self-PrP peptides are immunogenic in mice and suggest that this immune response might affect PrP-scrapie levels in certain conditions. PMID- 11477547 TI - Failure of MHC class II expression in neonatal alveolar macrophages: potential role of class II transactivator. AB - Neonatal peritoneal and blood macrophages are known to be ineffective in antigen presentation functions, and this manifests as inefficient MHC class II expression in response to IFN-gamma. The underlying mechanisms responsible for this maturational deficiency have not been elucidated. We show here that MHC class II expression in alveolar macrophages (AM) from neonates is also refractory to IFN gamma stimulation. Furthermore, by examining the intracellular pathway leading to MHC class II expression, we demonstrate that the site of the impairment is at the level of transcription. Thus, expression of mRNA encoding the class II transactivator (CIITA), MHC class II (RT1.B) and invariant chain (Ii) was low or undetectable in neonatal AM stimulated with concentrations of IFN-gamma that induced adult AM to up-regulate MHC class II expression. The failure of AM from young animals to express MHC class II was not due simply to deficient IFN-gamma receptor function since IFN-gamma-responsive genes such as IRF-1, IRF-2 and IP-10 were up-regulated in a dose-dependent manner from animals of all ages investigated. Importantly, the responsiveness of neonatal AM to IFN-gamma, as determined by MHC class II expression, could be modulated to adult levels when pre-cultured in vitro. This suggests that microenvironmental factors operative in vivo may play a role in suppressing the expression of MHC class II in AM from young animals. We have investigated the role of type I interferons but did not find them to be responsible for the inability of AM from young animals to induce MHC class II in response to IFN-gamma. PMID- 11477548 TI - Overexpression of Pax5 is not sufficient for neoplastic transformation of mouse neuroectoderm. AB - The developmental control genes of the Pax family are essential for brain development. Several Pax genes are also involved in chromosomal translocations causing malignancies in humans, and Pax5 expression is deregulated in medulloblastomas. We have investigated whether Pax5 can induce tumors in the developing mouse brain. Primary mouse embryonic neuroectodermal cells were retrovirally transduced with mouse Pax5 and transplanted into the brain of syngeneic host mice. No tumors developed in 36 transplants after one year, and there were no alterations in the differentiation pattern of the neural transplants. We then generated transgenic mice expressing human Pax5 under control of the Engrailed-2 promoter, which is expressed in the cerebellar external granule cell layer and in medulloblastomas. Sustained expression was achieved in the cerebellum of transgenic animals throughout lifetime. Expression levels were similar to those observed in human medulloblastomas. Again, cerebellar morphogenesis was undisturbed, and no tumors arose. These results strongly argue against a dominant transforming activity of PAX5 in NEC and in cerebellar granule cell precursors of mice, and underline the restricted tissue specificity of PAX5 related oncogenesis. PMID- 11477549 TI - Tumor heterogeneity in small hepatocellular carcinoma: analysis of tumor cell proliferation, expression and mutation of p53 AND beta-catenin. AB - Most hepatocellular carcinomas (HCCs) first occur as well-differentiated HCCs, from which poorly differentiated HCC cells develop because of dedifferentiation. In this study, we try to clarify the changes of dedifferentiation and cell proliferative activity and their relationship in small HCCs (less than 3.0 cm in diameter) and try to learn the mechanism of these changes by analysing the expressions and genetic changes of proliferation-related genes p53 and beta catenin. Of 41 surgically resected small HCCs, 11 were identified to have tumor heterogeneity. DNA from the 11 small HCCs, consisting of 29 intratumoral lesions and 11 noncancerous liver tissues adjacent to HCCs, was extracted from paraffin embedded tissue sections. Exons 5-8 of p53 gene and exon 3 of beta-catenin gene were amplified by polymerase chain reaction and analyzed by direct sequence. The serial sections were also immunostained by anti-Ki-67, p53 and beta-catenin antibody. Immunohistochemistry showed that the p53 overexpression was significantly related to the proliferative activities as evaluated by Ki-67 immunostaining and to the histological differentiation. The expression of beta catenin was found to be heterogeneously distributed not only in various histological grades of the same tumor but also in areas of the same histological grade. p53 and beta-catenin gene mutations were detected in 1 tumor respectively, both of which were second primary HCCs and also recurred later. The p53 mutation showed the same mutation pattern in heterogeneous subpopulations. beta-catenin mutation was detected only in the less differentiated lesion but not in the well differentiated lesion of tumor. In conclusion, our findings suggest that there was histological heterogeneity in small but established HCC, which was accompanied by increased proliferative activity and p53 overexpression. The overexpression of beta-catenin may be related to the proliferative activity and dedifferentiation of HCC. PMID- 11477550 TI - Increased cytotoxic effects of photodynamic therapy in IL-6 gene transfected cells via enhanced apoptosis. AB - PDT has been reported to induce cancer cell expression of cytokines, such as IL-6 and TNF-alpha, but it has been unclear whether cytokine expression by cancer cells is directly related to the antitumor effect of PDT. We treated Lewis lung carcinoma (LLC) cells with a new photosensitizer, mono-L-aspartyl chlorin e6 (NPe6) and light from a diode laser and found that expression of the mRNA of IL 2, IL-6, and TNF-alpha was increased by NPe6-mediated-PDT 6 hr later. To elucidate the mechanism of the direct anti-tumor effect of cytokine expression, we examined the photosensitivity of cytokine-gene-transfected cells, namely LLC IL-2, LLC-IL-6, and LLC-TNF-alpha cells, by MTT assay. The IL-6 gene transfected, LLC-IL-6 cells were significantly more sensitive to cytotoxic effects than the parent LLC cells and other cytokine gene-transfected cells. This finding indicates that IL-6 expression modulates cellular sensitivity to PDT and that IL 2 and TNF-alpha expressions does not. In addition, the apoptosis of LLC-IL-6 cells induced by NPe6-PDT was greater than in the other cells as determined by DNA fragmentation and staining of apoptotic nuclei. Because IL-6 has been reported to induce apoptosis by downregulating expression of Bcl-2, we analyzed the expression of apoptosis-related Bcl-2, Bax, and cytochrome C by Western blot analysis. Decreased expression of Bcl-2 and cytochrome C was observed in both LLC cells and LLC-IL-6 cells. Bax protein increased in a time-dependent manner, and the ratio of Bax to Bcl-2 rose markedly after PDT in LLC-IL-6 cells. These results suggest that the increased sensitivity of LLC-IL-6 cells to PDT-induced cytotoxicity results from the high ratio of Bax to Bcl-2 in the IL-6-dependent apoptotic pathway. In conclusion, IL-6 expression plays a role in cellular sensitivity to PDT, and combination of IL-6 and PDT may provide a new strategy for cancer treatment. PMID- 11477551 TI - Inhibition of proteasome function induced apoptosis in gastric cancer. AB - The ubiquitin-proteasome pathway plays a critical role in the degradation of cellular proteins and cell cycle control. Dysregulating the degradation of such proteins should have profound effects on tumor growth and causes cells to undergo apoptosis. The aims of this study are to evaluate the ubiquitin-proteasome pathway in gastric cancer and the potential role of pharmacological inhibition of proteasome on induction of apoptosis in gastric cancer cells. Gastric cancer cell lines AGS (p53 wild-type) and MKN-28 (p53 mutant) were treated with proteasome inhibitor MG132. The results showed that MG132 inhibited cell proliferation in AGS and MKN-28 cells in a time- and dose-dependent manner. The inhibition of cell proliferation was caused by apoptosis which was also time- and dose-dependent. AGS cells were more responsive to MG132 than MKN-28 cells. Induction of apoptosis was preceded by the activation of caspase-3, as measured by a colorimetric caspase-3 cellular activity and Western blotting of the cleavage of caspase-3 and its substrate PARP. Activation of caspase-7 was also exhibited. In addition, z VAD-fmk, a broad spectrum caspase inhibitor, reversed apoptosis induced by MG132 in AGS and MKN28 cells. Although z-DEVD-fmk, a specific caspase-3 inhibitor, suppressed MG132-induced apoptosis in MKN28 cells, it only partially rescued the apoptotic effect in AGS cells. Caspase-3 activation was the result of release of cytochrome c from mitochondria into the cytosol, as a consequence of upregulation of bax. There were overexpressions of all the proteasome-related proteins p53, p21(waf1) and p27(kip1) at 4 hr after proteasome inhibition which was identified by the accumulation of ubiquitin-tagged proteins. This was accompanied by accumulation of cells at G(1) phase. Our present study suggests that inhibition of proteasome function in gastric cancer cells induces apoptosis and proteasomal inhibitors have potential use as novel anticancer drugs in gastric cancer. PMID- 11477552 TI - Role of HGF/c-met system in invasion and metastasis of oral squamous cell carcinoma cells in vitro and its clinical significance. AB - We examined the role of the hepatocyte growth factor (HGF)/c-met system on invasion and metastasis of oral squamous cell carcinoma (SCC) cells. In monolayer culture, exogenous HGF marginally affected the growth of oral SCC cells (BHY, HN, IH) and human gingival epithelial cells (GE). In type I collagen matrix, however, HGF significantly enhanced the invasive growth of the cancer cells (p < 0.05). We detected the expression of c-met (HGF receptor) mRNA in all of the cancer cells, but not in human gingival fibroblasts (GF). Oral SCC cells did not secret HGF protein into the medium, but GF secreted a large amount of HGF protein (15 ng/ml). Furthermore, HGF markedly enhanced the migration of cancer cells in a Transwell invasion chamber. Then, we examined the serum levels of HGF in oral SCC patients, or HGF concentrations in oral cancer tissues. Serum levels of HGF in the patients were significantly higher than those in healthy volunteers (p < 0.05). After initial treatment, all of the tumor-free survivors showed a marked decline in the serum HGF levels. Furthermore, HGF concentrations in metastatic cancer tissues were significantly higher than those of non-metastatic cancer tissues and normal gingiva (p < 0.01). These results suggest that HGF plays an important role in invasion and metastasis of oral SCC cells as a paracrine factor, and an elevated HGF level in the cancer tissue can be a predictive marker for metastasis formation in patients with oral SCC. PMID- 11477553 TI - Cyclooxygenase inhibitors retard murine mammary tumor progression by reducing tumor cell migration, invasiveness and angiogenesis. AB - Tumor-derived prostaglandins (PGs) have been implicated in the progression of murine and human breast cancer. Chronic treatment with a non-selective PG inhibitor indomethacin was shown in this laboratory to retard the development and metastasis of spontaneous mammary tumors in C3H/HeJ female retired breeder mice. The present study examined the role of endogenous prostaglandins in the proliferation/survival, the migratory and invasive behavior and angiogenic ability of a highly metastatic murine mammary tumor cell line, C3L5, originally derived from a C3H/HeJ spontaneous mammary tumor. This cell line was shown to express high levels of cyclooxygenase (COX) -2 mRNA and protein as detected by Northern and Western blotting as well as immunostaining. PGE(2) production by C3L5 cells was primarily owing to COX-2, since this was blocked similarly with non-selective COX inhibitor indomethacin and selective COX-2 inhibitor NS-398, but unaffected with the selective COX-1 inhibitor valeryl salicylate. C3L5 cell proliferation/survival in vitro was not influenced by PGs, since their cellularity remained unaffected in the presence of PGE(2) or NS-398 or PG receptor (EP1/EP2) antagonist AH6809; a marginal decline was noted only at high doses of indomethacin, which was not abrogated by addition of exogenous PGE(2). Migratory and invasive abilities of C3L5 cells, as quantitated with in vitro transwell migration/invasion assays, were inhibited with indomethacin or NS-398 or AH6809 in a dose-dependent manner; the indomethacin and NS-398-mediated inhibition was partially reversed upon addition of exogenous PGE(2). An in vivo angiogenesis assay that used subcutaneous implants of growth factor-reduced matrigel inclusive of tumor cells showed a significant inhibition of blood vessel formation in these implants in animals treated with indomethacin compared with animals receiving vehicle alone. These studies show that selective and nonselective COX-2 inhibitors retarded tumor progression in this COX-2-expressing murine mammary tumor model by inhibiting tumor cell migration, invasiveness and tumor-induced angiogenesis. The inhibitory effects were not entirely PG dependent; some PG-independent effects were also noted. PMID- 11477554 TI - Differential regulation of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 expression in co-cultures of prostate cancer and stromal cells. AB - Tumor-stromal interactions have been suggested to be a critical factor in both tumor invasion and tumor metastasis. Here, we examined the role of tumor-stromal interactions using co-cultures of prostate cancer (PC) cells derived from primary and metastatic tumors with primary or immortalized stromal (fibroblast and smooth muscle) cells and their effect on matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) expression. Co-cultures of PC and stromal cells showed enhanced levels of pro-MMP-9 and reduced levels of TIMP-1 and TIMP 2. Whereas enhanced expression of pro-MMP-9 occurred in PC cells, the TIMPs were down-regulated in stromal cells. Induction of pro-MMP-9 and reduction of TIMP expression did not require cell-cell contact and were mediated by a soluble factor(s) present in the conditioned medium of the effector cell. Collagen I is a potent inducer of pro-MMP-9 in PC cells. Consistently, preliminary characterization of the soluble factor in the fibroblast conditioned medium revealed m.w. of approximately 100 to 250 kDa, and its effect on pro-MMP-9 expression was partly inhibited by an anti-alpha2 integrin antibody, a major collagen I receptor. Expression of pro-MMP-9 protein and mRNA was also induced in metastatic PC-3 cells grown in human fetal bone implants in SCID mice. Together, these findings demonstrate the importance of tumor-stromal interactions in the regulation of MMP and TIMP expression and their potential role in PC progression. PMID- 11477555 TI - Overexpression of homeobox gene HOXD3 induces coordinate expression of metastasis related genes in human lung cancer cells. AB - Homeobox-containing genes are expressed in spatiotemporal fashion during embryogenesis and act as master transcription-regulating factors which control the expression of a variety of genes involved in morphogenesis. They are also expressed in a tissue-specific manner in normal adult tissues and appear to give cells spatial information in the maintenance of their architectural integrity. We transfected a HOXD3 class I homeobox-containing gene into human lung cancer A549 cells and investigated alterations in gene expressions and phenotypes related to the maintenance of tissue architecture in HOXD3-overexpressing A549 cells. In the HOXD3-overexpressing cell lines, expression of E-cadherin was lost and plakoglobin was strongly repressed, whereas integrin alpha3 and beta3 were up regulated and N-cadherin and integrin alpha4 were newly expressed. Compared with parental and control transfectant lines, the HOXD3-overexpressing cell lines showed highly motile and invasive activity. Blocking experiments using anti integrin beta1 and beta3 suggested that the increased haptotaxis of the HOXD3 overexpressing cells to vitronectin resulted from increased expression and activation of integrin alphavbeta3, and that overexpression of the HOXD3 gene converted the integrin beta1-dependent haptotaxis to fibronectin into both integrin beta1- and beta3-dependent one. HOXD3 overexpression increased production of matrix-degrative enzymes including matrix metalloproteinase-2 and urokinase-plasminogen activator. When the tumor cells were intravenously injected into the tail veins of nude mice, HOXD3 transfectants formed a significantly large number of metastatic foci in lungs compared with the control transfectants. These findings suggest that HOXD3 can act as a metastasis-promoting gene in human lung cancer A549 cells. PMID- 11477556 TI - Induction of apoptosis in human colon carcinoma cells HT29 by sublethal cryo injury: mediation by cytochrome c release. AB - Cryosurgery is an emerging treatment for human solid tumors, notably colorectal liver metastasis. Cryosurgical procedures generate a thermal gradient of from at least -50 degrees C at the center of the tumor being treated to about 0 degrees C at the periphery. Cell death occurs by necrosis in the center, while the peripheral zone of frozen tumor harbors a mix of viable and dead tissue. In order to understand the mechanisms of cell death and survival in this peripheral area at risk for tumor recurrence, we have established an in vitro freezing system that mimics in vivo conditions of sublethal injury. HT29 colon cancer cells were subjected to freezing temperatures from -6 degrees C to -36 degrees C, thawed at room temperature for 30 min and rewarmed at 37 degrees C for a period of time. Post-freeze-thaw, cryolytic cells were evaluated by trypan blue exclusive assay. We also identified apoptotic cells after rewarming by cell shrinkage, nucleic condensation, TUNEL assay, DNA fragmentation and PARP degradation. The intensity of cryolysis and apoptosis was increased by lowering the freezing temperature. At -36 degrees C, all cells were dead immediately after freeze-thaw. A kinetic analysis of cryo-induced apoptosis showed that the commitment to enter apoptosis occurred right after the freeze-thaw period and lasted less than 8 hr after rewarming. We further demonstrated that freezing triggers one of the caspase cascade involved in apoptosis: release of cytochrome c from mitochondria to cytosol, followed by activation of caspase-9 and degradation of PARP. These results indicate the death of cancer cells under cryo-treatment at sublethal freezing temperature can be attributed 2 different modes, cryolysis as well as apoptosis. HT29 cells carrying p53 mutant have very quick response for induction of apoptosis by cryo-treatment and contain an intact pathway of caspase cascade. Further studies will address if mechanisms in cells with wild-type p53 will differ. PMID- 11477557 TI - High frequency of mitochondrial DNA mutations in glioblastoma multiforme identified by direct sequence comparison to blood samples. AB - In an earlier study, we showed that heteroplasmy in the mitochondrial genome of gliomas sometimes occurs in a D-loop polycytosine tract. We extended this study by pairwise comparisons between glioma samples and adjacent brain tissue of 55 patients (50 glioblastomas, 1 astrocytoma WHO grade III, 4 astrocytomas WHO grade II). We used a combination of laser microdissection and PCR to detect and quantify variations in the polycytosine tract. New length variants undetectable in the adjacent brain tissue were observed in 5 glioblastomas (9%). In 2 of these cases, samples from a lower tumor stage (WHO grade II) could be analyzed and revealed the early occurrence of these mutations in both cases. Since the mitochondrial D-loop contains additional repeats and highly polymorphic non coding sequences, we compared 17 glioblastomas with the corresponding blood samples of the same patients by direct sequencing of the complete D-loop. In 6 of these tumors (35%), instability was detected in 1 or 2 of 3 repeat regions; in 1 of these repeats, the instability was linked to a germline T-to-C transition. Furthermore, of 2 tumors (12%) 1 carried 1 and the other 9 additional transitions. In the latter patient, 6.7 kb of the protein coding mtDNA sequence were analyzed. Six silent transitions and 2 missense mutations (transitions) were found. All base substitutions appeared to be homoplasmic upon sequencing, and 89% occurred at known polymorphic sites in humans. Our data suggest that the same mechanisms that generate inherited mtDNA polymorphisms are strongly enhanced in gliomas and produce somatic mutations. PMID- 11477558 TI - Efficient antitumor immunity derived from maturation of dendritic cells that had phagocytosed apoptotic/necrotic tumor cells. AB - Dendritic cells (DCs) that acquired antigen from apoptotic tumor cells are able to induce major histocompatibility complex (MHC) class I-restricted cytotoxic T lymphocytes and antitumor immunity. In the present study, we investigated the efficiency of antitumor immunity derived from DCs that had phagocytosed apoptotic/necrotic BL6-10 melanoma cells compared with that of DCs pulsed with the tumor mTRP2 peptide. Our data showed that phagocytosis of apoptotic/necrotic tumor cells resulted in maturation of DCs with up-regulated expression of proinflammatory cytokines [interleukin (IL)-1beta, IL-6, tumor necrosis factor alpha, interferon-gamma and granulocyte-macrophage colony-stimulating factor], chemokines (MIP-1alpha, MIP-1beta and MIP-2), the CC chemokine receptor CCR7 and the cell surface molecules (MHC class II, CD11b, CD40 and CD86), and down regulated expression of the CC chemokine receptors CCR2 and CCR5. These mature DCs displayed enhanced migration toward the CC chemokine MIP-3beta in a chemotaxis assay in vitro and to the regional lymph nodes in an animal model in vivo. Our data also showed that vaccination with DCs that had phagocytosed apoptotic/necrotic BL6-10 cells was able to (i) more strongly stimulate allogeneic T-cell proliferation in vitro, (ii) induce an in vivo Th1-type immune response leading to more efficient tumor-specific cytotoxic CD8(+) T-cell mediated immunity and (iii) eradicate lung metastases in all 6 vaccinated mice compared with mice vaccinated with DCs pulsed with the tumor mTRP2 peptide, in which lung metastases were reduced (mean number of 16 per mouse) but not completely eradicated. Therefore, DCs that had phagocytosed apoptotic/necrotic tumor cells appear to offer new strategies in DC cancer vaccines. PMID- 11477559 TI - Detection and quantification of CD8(+) T cells specific for HLA-A*0201-binding melanoma and viral peptides by the IFN-gamma-ELISPOT assay. AB - Blood lymphocytes from HLA-A*0201-subtyped melanoma patients and healthy controls were screened for the presence of T cells specific for HLA-A*0201-binding melanoma and viral peptide antigens by the enzyme-linked immunoSPOT (ELISPOT) assay. CD8(+) cells were tested for peptide-specific IFN-gamma release immediately after selection as well as after 2 weeks of in vitro stimulation. After in vitro stimulation, CD8(+) T cells specific for influenza were measured in all patients and controls, whereas these T cells could be detected among nonstimulated CD8(+) cells in only 52% of individuals. Similarly, T cells specific for EBV were more frequently measured among in vitro-stimulated than nonstimulated CD8(+) cells. In nonstimulated CD8(+) cells, T cells specific for MART-1/Melan-A, gp100, tyrosinase and CAMEL were present in 4 (33%), 1 (8%), 1 (8%) and 3 (25%) of 12 patients, respectively. Only MART-1/Melan-A-specific CD8(+) T cells were found in 1 (11%) of 9 healthy controls. CD8(+) T cells specific for MAGE-2 were not observed. After in vitro stimulation, CD8(+) T cells specific for MART-1/Melan-A could be demonstrated in 6 (46%) of 13 patients and 2 (20%) of 10 controls. CD8(+) T cells specific for gp100 were detected in 1 patient after in vitro stimulation. No CD8(+) T cells specific for tyrosinase, MAGE-2 or CAMEL could be measured after in vitro stimulation. These data show that the ELISPOT assay allows direct ex vivo detection of CD8(+) T cells specific for viral and melanoma antigens. Furthermore, the data show that the sensitivity of the ELISPOT assay to measure influenza- and EBV-specific CD8(+) T cells can be enhanced by a short in vitro stimulation step, whereas opposing effects on numbers of CD8(+) T cells specific for melanoma antigens have been observed. PMID- 11477560 TI - Humanization and characterization of the anti-HLA-DR antibody 1D10. AB - 1D10 is a previously described antibody that binds to cells from a majority of B cell malignancies. The current studies were designed to further evaluate the antigen specificity of 1D10 and its potential as an immunotherapeutic agent. Studies with transfectants and immunoprecipitation demonstrated that 1D10 recognizes some, but not all, of the human HLA-DR beta chains. Both normal and malignant B cells can express the 1D10 antigen. A humanized version of 1D10 was produced using CDR grafting. The resulting antibody has an affinity that is similar to that of the parental murine antibody. In addition, the humanized antibody is capable of inducing complement-mediated cytotoxicity, antibody dependent cell cytotoxicity, and direct apoptosis of 1D10-expressing B cells. Based on these in vitro anti-tumor activities, we conclude humanized 1D10 deserves further evaluation as an immunotherapeutic agent. PMID- 11477561 TI - MHC class I negative phenotype of disseminated tumor cells in bone marrow is associated with poor survival in R0M0 breast cancer patients. AB - Changing the major histocompatibility complex (MHC) class I phenotype is a pivotal strategy of tumor cells to circumvent an effective immune response and is associated with tumor progression in cancer patients. Epithelial cells in bone marrow have been detected in various tumor types, but the clinical observation that only a portion of the patients with a positive bone marrow status develops solid bone metastasis suggests a certain molecular equipment of the isolated tumor cells as a prerequisite for metastatic formation. In the present study the prognostic impact of the MHC class I phenotype of disseminated epithelial cells in bone marrow was evaluated in a cohort of 30 curatively resected (R0) patients without distant metastases (M0) (designated R0M0) who had minimal residual disease. Immunocytochemical analysis using the alkaline/anti-alkaline immunogold double staining procedure revealed a heterogeneous MHC class I expression profile [monoclonal antibody (mAb) W6/32] of the epithelial cells (mAb CK2). In 16 patients (53.3%) all epithelial cells were human leukocyte antigen (HLA) class I positive (CK2+//W6/32+ phenotype). Eight patients (26.7%) showed complete loss of the HLA class I molecules (CK2+//W6/32- phenotype) and in 6 patients (20%) partial loss of HLA class I expression was found (CK2+//W6/32+ and - phenotype). CK2+ cells with the HLA class I negative phenotype (CK2+//W6/32- phenotype and CK2+//W6/32+ and - phenotype) were often derived from poorly differentiated (G3) primary breast carcinomas (p = 0.036) and were associated with short survival of the R0M0 patients (follow-up 15-98 months, log rank p = 0.072). These findings support the necessity to develop immmunotherapeutic strategies leading to the restoration of MHC class I positive phenotype. PMID- 11477562 TI - Resistance to topoisomerase poisons due to loss of DNA mismatch repair. AB - Sporadic breast carcinomas demonstrate microsatellite instability, reflecting the presence of DNA mismatch repair-deficient cells, in about one fourth of cases at the time of diagnosis. Loss of DNA mismatch repair has been reported to result in resistance not only to cisplatin and alkylating agents but also to the topoisomerase II poison doxorubicin, suggesting an association between DNA mismatch repair and topoisomerase II poison-induced cytotoxicity. Our study investigates the relationship between loss of MSH2 or MLH1 function and sensitivity to the topoisomerase I and II poisons, and to the taxanes, 2 classes of cytotoxic drugs commonly used in breast cancer. Two pairs of cell lines proficient and deficient in mismatch repair due to loss of either MSH2 or MLH1 function were used. Loss of either MSH2 or MLH1 function resulted in resistance to the topoisomerase II poisons doxorubicin, epirubicin and mitoxantrone, whereas only loss of MLH1 function was associated with low-level resistance to the topoisomerase I poisons camptothecin and topotecan. In contrast, there was no resistance to docetaxel and paclitaxel. Our data support the hypothesis that both MSH2 and MLH1 are involved in topoisomerase II poison-mediated cytotoxicity, whereas only MLH1 is involved in topoisomerase I poison-mediated cytotoxicity. Since our study shows that loss of DNA mismatch repair does not result in resistance to the taxanes, these drugs can be recommended for use in breast cancer deficient in mismatch repair. PMID- 11477563 TI - Cisplatin-induced apoptosis of mesothelioma cells is affected by potassium ion flux modulator amphotericin B and bumetanide. AB - Chemotherapeutic anti-cancer drugs induce cell death by the process of apoptosis. Efflux of potassium ions (K(+)) is necessary for cell volume reduction during apoptosis and increased inward pumping of K(+) thus counteracts apoptosis. Potassium flux modulation could therefore interact with apoptosis and affect the efficiency of cancer chemotherapeutics. We explored if the K(+) efflux stimulator amphotericin B, with or without the Na(+), K(+), 2Cl(-)-cotransport (K(+) influx) blocker bumetanide, could affect cisplatin- and carboplatin-induced apoptosis and cytotoxicity in the pulmonary mesothelioma cell line (P31). Apoptosis was determined by quantifying free nucleosomes and caspase-3 activity, and cytotoxicity was determined by clone formation and a fluorometric assay. The pan caspase enzyme inhibitor Boc-D-FMK was used to further determine the role of caspase activity in K(+)-flux-modulated cisplatin-/carboplatin-induced apoptosis and cytotoxicity. Amphotericin B (3.2 micromol/L) combined with bumetanide (100 micromol/L) potentiated cisplatin-induced free nucleosome and caspase-3 activity. The combination of the K(+) modulators did not, however, increase cisplatin cytotoxicity. The caspase inhibitor Boc-D-FMK, but unexpectedly also bumetanide, markedly reduced cisplatin cytotoxicity and annihilated the augmented cytotoxicity of cisplatin in the presence of amphotericin B. Carboplatin cytotoxicity was reduced by bumetanide, but not affected by amphotericin B. Carboplatin and carboplatin/bumetanide cytotoxicity was further reduced by Boc-D FMK. We conclude that the ability of cisplatin, and to a lesser extent carboplatin, to induce apoptosis is indeed influenced by cellular potassium flux modulators. We suggest that K(+) ionophores such as amphotericin B, and K(+) influx blockers such as bumetanide, alone or in combination, should be further evaluated for their potential clinical usefulness in influencing tumor cell apoptosis induced by cisplatin and other cancer chemotherapeutics. PMID- 11477564 TI - Association between expression of the MRP3 gene and exposure to platinum drugs in lung cancer. AB - To investigate the roles played by the multidrug resistance-associated protein (MRP1) homologues MRP3 and MRP4 in resistance to platinum drugs, we examined steady-state levels of mRNA for both MRP3 and MRP4 in normal lung and lung cancer specimens as well as peripheral mononuclear cells (PMN) after platinum drug exposure. MRP3 and MRP4 gene expression levels were monitored in the PMN of 10 previously untreated lung cancer patients within 24 hr after carboplatin (CBDCA) administration. Expression levels for both genes were also examined in 80 autopsy samples (40 primary tumors and 40 corresponding normal lung tissues) from 40 patients with lung cancer. MRP3 and MRP4 gene expression levels were assessed by quantitative reverse transcription-polymerase chain reaction. MRP3 expression levels in the PMN rose rapidly within 24 hr after administration of CBDCA, whereas MRP4 expression levels remained the same. Furthermore, MRP3 expression levels in normal lung and tumor tissues from autopsy samples that had been exposed to platinum drugs while the patients were alive were significantly higher than those in unexposed tissues, but again MRP4 expression levels remained the same. These results suggest that platinum drugs and/or the physiological stress response to xenobiotics induce expression of the MRP3 gene. PMID- 11477565 TI - Selective tumor sensitization to taxanes with the mAb-drug conjugate cBR96 doxorubicin. AB - The chimeric monoclonal antibody cBR96 conjugated to doxorubicin (cBR96-Dox) is selectively internalized by a wide variety of human carcinomas expressing an extended form of Lewis Y antigen (Le(y)). Endocytosis is followed by cleavage and release of free doxorubicin from the endocytic vesicles and subsequent cytotoxicity. Combination studies with standard anti-cancer agents, undertaken to further increase the potency of this targeted therapy, identified significant synergistic anti-tumor activity of cBR96-Dox and either of the taxanes paclitaxel or docetaxel. Treatment with cBR96-Dox 24 hr prior to paclitaxel resulted in a steady increase in the percentage of G(2) tumor cells and corresponding increase in sensitivity to taxanes. Cell cycle analysis indicated the cBR96-delivered doxorubicin was most effective against S-phase cells, yet cells exposed to even subtoxic levels progressed to and arrested in G(2), at a point of high sensitivity to the anti-tubulin agent paclitaxel. The synergy obtained by staged combination of cBR96-Dox and paclitaxel in vitro was reflected in significant anti-tumor efficacy in vivo against xenograft models of human lung and breast tumors that could not be achieved by either agent alone. The staged combination elicited significant or complete regressions of established human Le(y)-positive tumor xenografts using significantly reduced drug levels. Taken together, these data demonstrate a mechanistic approach to the selective elimination of Le(y) positive tumors by using targeted doxorubicin followed by taxane treatment. PMID- 11477566 TI - Dietary intake and microsatellite instability in colon tumors. AB - Microsatellite instability (MSI) occurs in approximately 15% of colon tumors. Other than relatively rare mutations in mismatch repair genes, the causes of MSI are not generally known. The purpose of this study was to determine if dietary intake of nutrients previously reported as being associated with colon cancer relate specifically to the MSI disease pathway. Data from a population-based case control study of adenocarcinoma of the colon were used to evaluate associations between dietary intake and MSI. Participants were between 30 and 79 years of age at time of diagnosis and included both men and women. Dietary intake data were obtained from a computerized diet history questionnaire. MSI was evaluated in several ways: by a panel of 10 tetranucleotide repeats, and by 2 mononucleotide repeats, BAT-26 and TGFbetaRII. A total of 1,510 cases had valid study data and tumor DNA on which we were able to obtain MSI status. Cases with and without MSI were compared with dietary data reported by 2,410 population-based controls to determine dietary associations that may be different for these 2 subsets of cases. We compared dietary intake for cases with and without MSI to further determine associations that are specific to the MSI disease pathway. When comparing MSI+ to MSI- tumors we observed that long-term alcohol consumption, especially intake of liquor, increased the probability of having a tumor with MSI [odds ratio (OR) for MSI+ vs. MSI- tumors for alcohol 1.6, 95% confidence interval (CI) 1.0-2.5; OR for liquor 1.6, 95% CI 1.1-2.4]. The likelihood of having MSI in the tumor from the combined effects of high alcohol consumption and smoking cigarettes showed a 70% excess in risk from the additive model. There were some suggestions that high intakes of refined grain might also be associated with MSI+ tumors, although associations were less consistent. Risk estimates for most other dietary factors did not differ substantially by MSI status. Data from this large population-based case-control study of colon cancer indicate that alcohol consumption, especially consumption of liquor, may increase the odds of an MSI+ tumor. Most other dietary factors do not appear operate exclusively in the MSI+ disease pathway. PMID- 11477567 TI - Psychological consequences of predictive genetic testing for hereditary non polyposis colorectal cancer (HNPCC): a prospective follow-up study. AB - Predictive genetic testing for cancer allows identification of those with the mutation (mutation positive) who should undergo cancer surveillance aiming at early detection of cancer and those without the mutation (mutation negative), whose unnecessary worry can be alleviated and who need not undergo frequent surveillance. However, there is a risk that predictive testing might have a harmful emotional impact on an individual. In the course of a predictive genetic testing protocol, we assessed general anxiety (by the State-Trait Anxiety Inventory [STAI]), fear of cancer and death, satisfaction with life and attitude to the future using a questionnaire survey in 271 individuals tested for hereditary non-polyposis colorectal cancer (HNPCC). Measurements were made before the first counseling (baseline), at the test disclosure session (STAI only) and 1 and 12 months after disclosure. Although at every measurement, the mutation positive individuals were more afraid of cancer than those who were mutation negative, in both groups fear of cancer decreased significantly from baseline after disclosure. The mutation-positive subjects were more anxious than their counterparts immediately after the test disclosure, but the differences had disappeared at the follow-ups. In other variables, neither differences between the groups defined by mutation status nor changes with time were detected. Our findings suggest that counseling and testing relieve fear of cancer; no harmful emotional impact was detectable at the 1-year follow-up. To confirm these findings, however, the impact of testing should be studied after a longer interval. Furthermore, to evaluate the ultimate interpretation of these results, studies are needed to investigate the impact of fear of cancer on surveillance behavior among the mutation-positive subjects. PMID- 11477568 TI - Contralateral breast cancer in Vaud, Switzerland. PMID- 11477569 TI - Drug-resistance in human melanoma. AB - Advanced malignant melanoma has a poor prognosis since chemotherapy is mostly ineffective due in part to the intrinsic and/or extrinsic resistance of melanoma cells to systemic treatment with anti-neoplastic agents. The reasons for the chemoresistant phenotype are unknown. The relevance of well-analyzed drug resistance mechanisms, e.g., intracellular/extracellular transport and induction of certain enzyme systems, is reviewed. Most anti-cancer drugs kill susceptible cells through induction of apoptosis. Therefore, it appears that differences in the apoptotic pathways which lead to apoptotic deficiency may account for the ability of some tumor cells to resist drug therapy. Human melanomas, which are characteristically drug-resistant, are more likely to have altered apoptotic pathways and fewer pro-apoptotic molecules. Tumor cells with these characteristics are seldom sensitive to drugs. The complexity of the molecular variants involved in signal transduction along apoptotic pathways suggests that the cell may have a variety of possibilities for regulating apoptosis and generating apoptotic deficiency. Thus, apoptosis and apoptotic deficiency should be analyzed to better clarify the mechanisms of melanoma resistance. PMID- 11477570 TI - Binding of a SART3 tumor-rejection antigen to a pre-mRNA splicing factor RNPS1: a possible regulation of splicing by a complex formation. AB - We recently reported the identification of a human SART3 gene that encodes a tumor-rejection antigen recognized by cytotoxic T lymphocytes (CTLs). The squamous-cell carcinoma antigen recognized by T cells-3 (SART3) is an RNA-binding protein expressed in the nucleus of the majority of proliferating cells, including normal cells and malignant cells, but not in normal tissues except for the testes and fetal liver. To determine its biologic function, we employed a 2 hybrid screening in yeast for proteins interacting with SART3, and this method yielded a pre-mRNA splicing factor (RNA-binding protein prevalent during the S phase or RNA-binding protein with a serine-rich domain [RNPS1]) that activated both constitutive and alternative splicing of pre-mRNA in vitro. Interaction of SART3 with RNPS1 through the physical association of N-terminal domains of RNPS1 was confirmed by both in vitro pull-down assay and immunoprecipitation assay. Cotransfection of the 2 genes changed the distribution pattern of SART3 from diffuse nucleoplasmic spreading to nuclear speckled regions in which the RNPS1 was colocalized, suggesting a complex formation of the 2 proteins. In cooperation with RNPS1, SART3 stimulated the proximal alternative 3' splicing of a calcitonin dihydrofolate reductase chimeric minigene pre-mRNA. These results suggest that SART3 is involved in the regulation of mRNA splicing probably via its complex formation with RNPS1. PMID- 11477571 TI - p16(INK4a) Promoter hypermethylation of non-tumorous tissue adjacent to gastric cancer is correlated with glandular atrophy and chronic inflammation. AB - The p16(INK4a) tumor suppressor gene can be inactivated by promoter region hypermethylation in many tumor types including gastric cancers. However, p16(INK4a) promoter hypermethylation in the surrounding non-tumorous tissues of gastric cancers has not been studied in detail. We therefore examined 46 gastric cancers, corresponding adjacent non-tumorous tissue samples and 8 gastric tissue samples of chronic gastritis by performing methylation-specific polymerase chain reaction, and we analyzed p16(INK4a) protein expression using immunohistochemistry and Western blot. p16(INK4a) promoter hypermethylation was observed in 43% of gastric cancers and 59% of adjacent non-tumorous tissues; however, none of the samples retrieved from the chronic gastritis patients displayed p16(INK4a) promoter hypermethylation. Gastric cancers showed an inverse correlation between vascular invasion and p16(INK4a) promoter hypermethylation, and adjacent non-tumorous tissues displayed a close association among the grade of chronic inflammation, presence of glandular atrophy and p16(INK4a) promoter hypermethylation. p16(INK4a) expression was markedly decreased in samples with p16(INK4a) promoter hypermethylation when compared with samples without p16(INK4a) promoter hypermethylation. These results suggest that p16(INK4a) promoter hypermethylation is an early and frequent event in gastric carcinogenesis and may serve as a new prognostic biomarker for the risk of gastric cancers. PMID- 11477572 TI - Relation of the induction of epidermal ornithine decarboxylase and hyperplasia to the different skin tumor-promotion susceptibilities of protein kinase C alpha, delta and -epsilon transgenic mice. AB - To define the in vivo role of individual PKC isoforms in mouse skin carcinogenesis, we previously characterized FVB/n transgenic mice that over expressed epitope-tagged PKC delta (T7-PKC delta) or PKC epsilon (T7-PKC epsilon) isoforms under the regulation of the human K14 promoter. In continuation of our prior PKC isoform specificity studies, we now report the generation of FVB/n transgenic mice with K14-regulated, epitope-tagged PKC alpha (T7-PKC alpha). T7 PKC alpha transgenic mice (line 115) express 8-fold more PKC alpha protein than wild-type mice. Using high-resolution immunogold cytochemistry, we determined that transgenic over-expression of T7-PKC alpha did not alter the subcellular localization of PKC alpha but that the density of PKC alpha staining increased. PKC alpha localized primarily to the cytoskeleton (tonofilaments, tight junctions) and cell membranes, with modest but definite nuclear labeling also identified. Also, PKC alpha over-expression did not alter the immunoreactive protein levels of other PKC isoforms (delta, epsilon, eta, zeta, mu) in the epidermis. Skin tumor-promotion susceptibility was compared among all 3 lines of T7-PKC transgenic mice (alpha, delta and epsilon). While T7-PKC alpha had no effect on skin tumor promotion by TPA, T7-PKC delta reduced papilloma burden by 76% compared to wild-type controls. T7-PKC epsilon further reduced papilloma burden to 93% compared to wild-type controls but still resulted in the development of squamous-cell carcinoma. To find potential mechanisms of PKC associated differences in tumor promotion, the induction of known downstream effectors of tumor promotion, ornithine decarboxylase (ODC) activity and epidermal hyperplasia, was determined. Despite long-term papilloma inhibition in both PKC delta and PKC epsilon transgenic mice, the induction of ODC by TPA was not attenuated in PKC delta and epsilon mouse lines. Both PKC transgenic and wild type mice exhibited sustained hyperplasia after repeated TPA treatments. However, TPA-induced epidermal hyperplasia in T7-PKC epsilon mice was significantly increased (52%) compared with T7-PKC alpha, T7-PKC delta and wild-type mice. TPA induced ODC activity and the resultant accumulation of polyamines may play different roles (e.g., induction of apoptosis vs. proliferation) in the pathways leading to the induction of cancer in PKC alpha, PKC delta and PKC epsilon transgenic mice. PMID- 11477573 TI - Quantitative cell dispersion analysis: new test to measure tumor cell aggressiveness. AB - Tumor progression requires the dispersion of epithelial cells from neoplastic clusters and cell invasion of adjacent stromal connective tissue. Aiming at demonstrating the precise relationships between cell dispersion and cell invasion, related respectively to expression of E-cadherin/catenin complex and matrix metalloproteinases (MMPs), we developed an original in vitro model of cell dispersion analysis. Our study reports the validation of this model that allowed us to analyze and quantify the cell cohesion level by means of time-lapse videomicroscopy and computer analysis based on the observation of spatial and temporal cell distribution. Our model was able to distinguish 2 groups among different human bronchial and mammary epithelial cells previously characterized for the expression of E-cadherin/catenin complex and MMPs and their invasive capacity in the Boyden chamber assay. The first group (16HBE14o(-), MCF-7, T47D) that expressed membranous E-cadherin and beta-catenin, and was negative for MMP-2 expression and non-invasive, displayed a highly cohesive pattern corresponding to a cluster spatial distribution. The second group (Beas2B, BZR, BZR-T33, MDA-MB 231, MDA-MB-435, BT549 and HS578T) that was invasive and showed lack of expression of E-cadherin and a cytoplasmic redistribution of beta-catenin, displayed a dispersed pattern corresponding to a random spatial distribution. Downregulation of E-cadherin by a blocking antibody induced a more random distribution. Conversely, expression of E-cadherin by cDNA transfection induced a cluster distribution. Moreover, tumor cell lines that co-expressed MT1-MMP and MMP-2 (Beas2B, BZR, BZR-T33, MDA-MB-435, BT549 and HS578T) showed a more dispersed pattern than tumor cell lines that did not express MMP-2 (MDA-MB-231). In conclusion, we demonstrated that the spatial group behavior of cell lines, i.e., their cohesion/dispersion ability, reflects their invasive properties. Thus, this model of cell dispersion analysis may represent a new test to measure tumor cell aggressiveness. PMID- 11477574 TI - Novel inactivating mutations of transforming growth factor-beta type I receptor gene in head-and-neck cancer metastases. AB - Carcinoma cell lines are frequently refractory to transforming growth factor-beta (TGF beta)-mediated cell cycle arrest. Whether and how TGF beta signaling is disrupted in the majority of human tumors, however, remains unclear. To investigate whether TGF beta signaling might be disrupted by inactivation of the key signaling molecule, the TGF beta type I (T beta R-I) receptor, and whether or not T beta R-I inactivation is associated with late stage disease, we conducted a comprehensive structural analysis of the T beta R-I gene in fine-needle aspirates of 23 head-&-neck cancer metastases. We encountered 4 different mutations of T beta R-I, 3 of which have not been previously identified. In 1 case, we found a somatic intragenic 4-bp deletion predicting for a truncation of the receptor protein. This is the first example of a true loss-of-function mutation of T beta R-I in a human epithelial neoplasm. In 2 other cases, we identified missense mutations located between the juxtamembrane- and serine-threonine kinase domains. One of these resulted in an alanine-to-threonine substitution (A230T), which disrupts receptor signaling activity by causing rapid protein degradation within the endoplasmatic reticulum. This represents a novel mechanism of inactivation of a TGF beta signaling intermediate. Finally, we identified a serine-to-tyrosine substitution at codon 387 (S387Y) in a metastasis but not in the corresponding primary tumor. We had previously shown this S387Y mutant to be predominantly associated with breast cancer metastases and to have a diminished ability to mediate TGF beta-dependent signaling. In aggregate, these findings provide further support for the hypothesis that inactivation of the TGF beta signaling pathway occurs in a significant subset of human cancers. PMID- 11477575 TI - Clinicopathological significance of vascular endothelial growth factor (VEGF)-C in human esophageal squamous cell carcinomas. AB - The purpose of this study was to investigate the expression of vascular endothelial growth factor (VEGF) -C in human esophageal squamous cell carcinomas to elucidate its role in lymph node metastasis and tumor progression. The expression of VEGF-C and flt-4 genes was examined in 5 esophageal carcinoma cell lines, 12 fresh biopsy specimens and 48 archival surgical specimens of human esophageal carcinoma tissues by RT-PCR and immunohistochemistry. Immunohistochemistry using antibodies against CD34 (endothelial cell specific) was also carried out and microvessels were quantified by counting vessels in a 200x field in the most vascular area of the tumor. Of the 5 human esophageal carcinoma cell lines, 4 constitutively expressed VEGF-C mRNA. In 8 (66.7%) of 12 cases, VEGF-C mRNA was detected in only tumor tissues but not in normal mucosa by RT-PCR. There was a significant relationship between VEGF-C and flt-4 mRNA expression. Out of the 48 surgical specimens of esophageal carcinomas, 19 (39.6%) and 10 (20.8%) exhibited intense VEGF-C immunoreactivity in the cytoplasm of many cancer cells and the stromal cells, respectively. In contrast, Flt-4 was mainly expressed on the lymphatic endothelial cells. Normal and dysplastic esophageal squamous epithelium exhibited no or faint cytoplasmic staining of VEGF-C. VEGF-C expression correlated with depth of tumor invasion, tumor stage, venous invasion, lymphatic invasion and lymph node metastasis. Vessel count was significantly higher in the VEGF-C positive tumors than in the negative tumors. These results overall suggest that VEGF-C may play a role in tumor progression via lymphangiogenesis and angiogenesis in human esophageal carcinoma. PMID- 11477576 TI - The E-cadherin gene is silenced by CpG methylation in human oral squamous cell carcinomas. AB - Reduction of E-cadherin strongly relates to invasiveness and metastasis in vitro. To clarify CpG methylation around the promoter region of the E-cadherin gene in oral squamous cell carcinoma (SCC), we examined the DNA samples of various human SCC cell lines and primary oral SCC tissues by methylation-specific polymerase chain reaction (MSP). CpG methylation of the E-cadherin gene markedly correlated to the reduction of E-cadherin expression in human oral SCC cell lines. In primary oral SCC tissues, only 1 of 5 preserved E-cadherin-expressing tissues was methylated, whereas methylation was found in 17 (94.4%) of 18 E-cadherin-reduced tissues. Our results suggest that reduction of E-cadherin expression is associated with CpG methylation of the E-cadherin gene promoter. We recently established two cell lines with high and low metastatic potential, UM1 and UM2, from SCC primary tongue tissue of a patient. E-cadherin expression of high metastatic UM1 was clearly lower than that of low-metastatic UM2, and MSP results showed CpG methylation in the UM1 but not the UM2 cell line. To investigate whether demethylation of CpG methylation of the E-cadherin gene could restore expression and function of E-cadherin, we treated UM1 with the demethylating agent 5-azacytidine (5-aza) and found that E-cadherin expression was indeed restored by demethylation. Moreover, in the demethylated UM1, invasion of the collagen gel was clearly suppressed compared with the untreated UM1. These results suggested that inactivation of E-cadherin expression resulted from CpG methylation of the gene promoter; a correlation between CpG methylation of the E cadherin gene promoter and invasive potential was also suggested. PMID- 11477577 TI - The IL-6 receptor antagonist SANT-7 overcomes bone marrow stromal cell-mediated drug resistance of multiple myeloma cells. AB - The bone marrow micro-environment produces a number of different survival factors that are important for the malignant growth and drug resistance of multiple myeloma (MM) cells. One of the main factors reported to be essential for survival and growth of MM cells in some experimental systems is IL-6. Therefore, the development and testing of substances that interfere with IL-6 or IL-6 receptor (IL-6R) function might have therapeutic value for the treatment of MM. We analyzed the effect of the IL-6R antagonist SANT-7 on growth and survival of the IL-6--dependent MM cell lines INA-6 and XG-1 as well as primary MM cells from 7 patients co-cultured with bone marrow stromal cells (BMSCs). In particular, we were interested in whether SANT-7 enhances the growth-inhibitory effects of dexamethasone (Dex) and all-trans-retinoic acid (ATRA). None of the drugs when tested as a single substance, including SANT-7, induced major growth inhibition if MM cells were co-cultured with primary human BMSCs. However, when Dex and ATRA were given in combination with SANT-7, strong growth inhibition was achieved in cell lines and primary MM cells. This effect was due to cell-cycle arrest and induction of apoptosis. PMID- 11477578 TI - Increased cyclooxygenase-2 (cox-2) expression and activity in a murine model of metastatic breast cancer. AB - Elevated prostaglandin E(2) (PGE(2)) production is a common feature of human malignancies. This activity has often been attributed to increased metabolic activity of the cyclooxygenase enzymes, although a direct comparison of these 2 parameters i.e., prostaglandin production and cox protein expression, is rarely performed in the same malignant tissue. Using a murine model of metastatic breast cancer, we show that PGE(2) levels are positively correlated with increased tumorigenic and metastatic potential. Because prostaglandin synthesis is a product of 2 isoforms of the cyclooxygenase enzyme, we examined the expression and activity of both isoforms. All tumor cell lines examined, regardless of phenotype, express both cox-1 and cox-2 proteins in vitro. In contrast to the uniform cox-2 expression in vitro, only tumors resulting from the transplantation of metastatic cell lines express cox-2 in vivo. Cox-1 is detected in both metastatic and nonmetastatic tumors. Thus, this is the first evidence that, in the tumor milieu, cox-2 expression can be regulated differently in metastatic vs. nonmetastatic lesions. Examination of PGE(2) synthesis in vitro reveals that nearly complete inhibition of prostaglandin synthesis occurs in the presence of either indomethacin, which inhibits both isoforms, or NS398, which is selective for the cox-2 isoform. Thus, even though cell lines express both isoforms, the majority of the prostaglandin synthesis stems from the activity of the inducible, cox-2 isoform. Likewise, cell growth is inhibited by both indomethacin and NS398 in a dose-dependent manner, albeit at higher drug concentrations than required to ablate PGE(2) synthesis. Despite the inhibition of prostaglandin synthesis, the cox-2 enzyme levels (protein and mRNA) were increased by either indomethacin or NS398. PMID- 11477579 TI - Inhibition of estrogen-dependent breast cell responses with phenylacetate. AB - The aromatic fatty acid phenylacetate (PA) and its analogs have come under intense investigation due to their ability to cause the growth arrest of a variety of neoplasia, including human breast cancer. We have determined that PA and its halide derivative 4-chlorophenylacetate (4-CPA) showed marked antiproliferative activity on 3 of 6 human breast cancer cell lines tested. Interestingly, the 3 cell lines that were growth inhibited by PA and 4-CPA were estrogen receptor (ER) positive (T47-D, MCF-7 and ZR-75-1) whereas those that were little affected by these compounds were ER-negative (MDA-MB-157, MDA-MB-231 and SK-Br-3). Dose response studies indicated that 4-CPA inhibited the growth of the sensitive (ER+) cell lines with a potency 3-4 times that of PA. These findings suggest that there is "cross-talk" between the PA and estrogen signaling pathways such that PA can directly inhibit estrogen-dependent events. This hypothesis was directly tested in vitro using ER+ MCF-7 cells that were stably transfected with a luciferase reporter construct driven by the full length (1745 bp) cyclin D1 promoter (MCF-7-D1). Our experiments with MCF-7-D1 cells indicated that PA and 4-CPA inhibited basal and estrogen-induced reporter gene activity by up to 90%, resulting in almost complete elimination of estrogen-dependent cyclin D1 gene activation. Using a reporter gene construct (ERE(V)-tk-Luc) containing a canonical estrogen response element that was transiently transfected into MCF-7 and MDA-MB-231 cells, we have also demonstrated inhibition of promoter activity by PA and 4-CPA that was directly mediated by blockage of activity through the ERE. Taken together, these findings indicate that PA analogs possess potent antiestrogen properties that may, at least partly, account for their antiproliferative effects on ER+ breast cancer cells. The data suggests a novel mechanism of action that might bypass some of the limitations of conventional antiestrogen therapy. PMID- 11477580 TI - Intravascular HBO(2) saturations, perfusion and hypoxia in spontaneous and transplanted tumor models. AB - Clinical trials utilizing strategies to manipulate tumor oxygenation, blood flow and angiogenesis are under way, although limited quantitative information exists regarding basic tumor pathophysiology. The current study utilized murine KHT fibrosarcomas, spontaneous mammary carcinomas and first-generation spontaneous transplants to examine heterogeneity in vascular structure and function, to relate these changes to the distribution of tumor hypoxia and to determine whether fundamental relationships among the different pathophysiological parameters exist. Three methods were included: (i) immunohistochemical staining of anatomical and perfused blood vessels, (ii) cryospectrophotometric measurement of intravascular oxyhemoglobin saturations and (iii) fluorescent detection of the EF5 hypoxic marker. While a distinct pattern of decreasing oxygenation with increasing distance from the tumor surface was observed for KHT tumors, striking intertumor variability was found in both spontaneous and first-generation transplants, with a reduced dependence on tumor volume. EF5 hypoxic marker uptake was also much more heterogeneous among individual spontaneous and first generation tumors compared to KHT. Although mammary carcinomas demonstrated fewer anatomical blood vessels than fibrosarcomas, the proportion of perfused vessels was substantially reduced in KHT tumors, especially at larger tumor volumes. Vascular morphology, tissue histological appearance and pathophysiological parameters differed substantially between KHT tumors and both spontaneous and first-generation tumors. Such differences in vascular structure and function are also likely to correlate with altered response to therapies targeted to the vascular system. Finally, spontaneous differentiation status, tumor morphology, vascular configuration and function were well preserved in first-generation transplanted tumors, suggesting a close relationship between vascular development and function in early-generation transplants and spontaneous tumor models. PMID- 11477581 TI - Superficial and metachronous invasive bladder carcinomas are clonally related. AB - Synchronous and metachronous tumors are frequently observed in the urinary tract and may be explained by the concept of 'field cancerization,' i.e., exposure to carcinogens leading to the independent transformation of many urothelial cells resulting in oncogenetically unrelated tumors. Increasing evidence, however, supports the concept of clonality, i.e., the progeny of a single transformed cell spreads through the urinary system resulting in genetically related tumors. The aim of our study was to investigate the putative clonal origin of invasive urothelial cell carcinomas (UCCs) of the bladder from a prior superficial tumor. We selected 6 patients (5 males and 1 female) with superficial and subsequent invasive UCC tumors. All patients were previously diagnosed with a p53 mutation in their invasive tumor. At least 1 superficial and 1 invasive tumor of the same patient were analyzed for mutations in the p53 tumor suppressor gene by PCR-SSCP and, in case of a band shift, followed by direct sequencing. In all patients the same p53 mutation was found in the superficial and subsequent invasive tumor(s). All tumors arose from the same progenitor cell. These results support the concept of a clonal origin of superficial and metachronous invasive bladder UCCs. PMID- 11477582 TI - Changes in p14(ARF) do not play a primary role in human chondrosarcoma tissues. AB - The locus encoding the tumor suppressor p16 has been found to code for a second, different protein. This protein, p14(ARF), has been shown to protect p53 from degradation. Like p16, its gene is often altered in different cancers. In this study, the first unique exon, exon 1 beta, of p14(ARF), has been studied in 22 chondrosarcoma tissues using polymerase chain reaction, DNA sequencing and methylation-specific polymerase chain reaction. One chondrosarcoma was found to have exon 1 beta homozygously deleted, but neither mutations nor methylations were found in any of the chondrosarcomas. This indicates that genetic changes of p14(ARF) are a rare event in chondrosarcoma. PMID- 11477583 TI - Adenoviral gene transfer into dendritic cells efficiently amplifies the immune response to LMP2A antigen: a potential treatment strategy for Epstein-Barr virus- positive Hodgkin's lymphoma. AB - The EBV-encoded LMP2A protein is consistently expressed in EBV(+) Hodgkin's lymphoma and can be targeted by CTLs. CTLs stimulated conventionally by LCLs have little activity against LMP2A(+) target cells. Here, we describe an alternative approach, based on the in vitro stimulation of CTLs with DCs genetically modified with 2 E1/E3-deleted recombinant adenoviruses, AdGFPLMP2A, encoding a fusion gene of GFP and LMP2A, and AdLMP2A, encoding LMP2A only. Transduction of DCs with AdGFPLMP2A at MOI 1,000 resulted in LMP2A expression in up to 88% of DCs. LMP2A protein was expressed in 40% of DCs transduced with AdLMP2A at an MOI of 100. Higher MOI resulted in DC death. CTL lines activated by transduced DCs had a higher frequency of LMP2A tetramer-specific CTLs than CTL lines activated by LCLs. CTLs stimulated with transduced DCs lysed both autologous fibroblasts infected with vaccinia virus LMP2A (FBvaccLMP2A) and autologous LCLs, which express LMP2A at lower levels. In contrast, CTLs generated from the same donors by stimulation with autologous LCLs showed minimal lysis of FBvaccLMP2A. Moreover, 1 donor who did not respond to LMP2A when CTLs were stimulated with LCLs became a responder when LMP2A was expressed by transduced DCs. Hence, recombinant adenoviruses encoding LMP2A effectively transduce DCs and direct the generation of LMP2A-specific CTLs. This approach will be a potent strategy in Hodgkin's lymphoma immunotherapy. PMID- 11477584 TI - EBNA promoter usage in EBV-negative Burkitt lymphoma cell lines converted with a neomycin-resistant EBV strain. AB - Latent Epstein-Barr virus (EBV) uses two alternative strategies to express the Epstein-Barr nuclear antigens (EBNAs). Resting normal B cells harboring latent virus and Burkitt's lymphoma (BL) cells use monocistronic messages generated from the Q promoter (restricted strategy). EBV-transformed immunoblasts express all EBNAs by using giant messages generated from the W/C promoter (full program). Whether the virus establishes the restricted program on primary infection of a BL cell (or its progenitor) or, alternatively, whether such cells are generated by phenotypic down-regulation from the immunoblast is unclear. We found previously that conversion of EBV-negative BL lines to EBV-positive sublines required repeated exposure to large virus doses. The converted sublines used the full program. However, the possibility that cells with a full program had a selective advantage during the long period of in vitro passage could not be excluded. We therefore infected EBV-negative BL lines with recombinant EBV carrying a neomycin resistance marker. Most convertants of the 12 lines tested were positive for YUK splicing, indicative of the full program, but some were also positive for the restricted QUK splice program. One convertant DG75 line showing both YUK and QUK was cloned and gave rise to stable QUK users. We conclude that EBV infection of established BL lines can give rise to subclones with either the full or the restricted program. The fact that all EBVs carrying BL lines use the restricted program in vitro may be a consequence of immunoselection. PMID- 11477585 TI - Photodynamic therapy of locoregional breast cancer recurrences using a chlorin type photosensitizer. AB - Chest wall recurrences are a frequent problem in patients treated by mastectomy for breast cancer. Surgery and ionizing radiation are established treatment modalities in these cases. Photodynamic therapy (PDT) provides an alternative treatment modality using a photosensitizer and laser light to induce selective tumor necrosis. PDT was performed as compassionate use in 7 patients aged 57.6 years (+/-12.6 SD). A total of 89 metastatic skin nodes were treated in 11 PDT sessions. As photosensitizer meta-tetra(hydroxyphenyl)chlorin (m-THPC) was applied intravenously. Patients (n = 3) photosensitized with a drug dose of 0.10 mg/kg bodyweight were irradiated 48 hr after drug application at a lightdose of 5 J/cm(2). Patients (n = 4) were illuminated by an optical dose of 10 J/cm(2) 96 hr after photosensitization with 0.15 mg/kg. Laser light at a wavelength of 652 nm was generated by a diode laser and applied by a front lens light diffuser using a fluence rate of 20--25 mW/cm(2). PDT using m-THPC resulted in complete response in all patients. Response to treatment did not differ when using the 2 different drugdose protocols. Healing time depended mainly on the size of the illumination field but not on the lightdose. Pain score usually raised 1 day after PDT and lasted at higher levels for about 10 days. Healing time usually ranged between 8- 10 weeks. Photodynamic technique offers a minimal-invasive, outpatient treatment modality for recurrent breast cancer on the chest wall with few side effects, high patient's satisfaction and with possible repetitive application. PMID- 11477586 TI - Glutathione-associated enzymes in head and neck squamous cell carcinoma and response to cisplatin-based neoadjuvant chemotherapy. AB - Glutathione S-transferases (GSTs) are metabolic phase II enzymes that promote reactive metabolite elimination by conjugating them to glutathione (GSH). Because of their important role in xenobiotic metabolism and detoxification, they have been implicated in carcinogenesis processes, especially epithelium transformation. Moreover, their influence on response to chemotherapy in cancer patients has been demonstrated. Genetic polymorphisms for GSTM1, GSTT1 and GSTP1 have been found in human populations and have been shown to have phenotypic consequences. To investigate the role of GST enzymes in carcinogenesis and in response to chemotherapy in patients with head and neck squamous cell carcinoma (HNSCC), GSTP1, GSTM1 and GSTT1 were studied prospectively in a large series of HNSCC patients. Correlations between GST alterations, p53 mutation status and clinical response to chemotherapy were investigated. We showed that the risk of developing laryngeal cancer was increased by 2.6-fold [95% CI 1.6--6.1] in patients with the GSTM1 null genotype and by 2.8-fold [95% CI 0.9--8.1] in patients with the homozygous GSTP1 val105 genotype. Furthermore, individuals with this latter genotype were over-represented in the p53 mutation group (p = 0.05). After storage duration and hemolysis adjustment, a significantly lower plasmatic GSTP1 level was observed in complete responders compared with partial and non responders (mean: 4.4 +/- 0.06 microg/l, 4.7 +/- 0.06 microg/l and 4.7 +/- 0.07 microg/l; p = 0.05), respectively. The prevalence of p53-mutated tumors was significantly higher in the group of non-responders (81%) compared with partial (60%) and complete responders (64%) (p = 0.05). Two types of multivariate analysis were performed including parameters that have been shown to influence response to chemotherapy significantly in univariate analysis. p53 mutations and high tumor stage are independent factors of non-response to chemotherapy, whereas plasmatic GSTP1 levels and low tumor stage are independent factors of complete response. Our data suggest that GST enzymes are associated with larynx cancer and that their use as predictive factors and treatment targets should be further explored. PMID- 11477587 TI - Mouse macrophage metalloelastase gene delivery by HVJ-cationic liposomes in experimental antiangiogenic gene therapy for murine CT-26 colon cancer. AB - We previously demonstrated that gene replacement of mouse macrophage metalloelastase (MME) into murine melanoma cells that grow rapidly and are MME deficient suppresses the primary tumor growth in vivo by halting angiogenesis. The aim of the present study was to evaluate the effectiveness of gene therapy against cancer using a cDNA-encoding MME gene. In a subcutaneous tumor model of CT-26 mouse colon cancer cells that are MME deficient, syngeneic mice repetitively treated with direct injections into the tumors of MME- hemagglutinating virus of Japan (HVJ), a type of HVJ-cationic liposome encapsulating a plasmid expressing MME, developed smaller tumors (210 +/- 47.2 mm(3) versus 925 +/- 156 mm(3) mean +/- SEM; p = 0.0004) with fewer microvessels (10.25 +/- 1.03 vs. 17.25 +/- 2.14; p = 0.03) than control mice. TUNEL staining revealed a significant increase of apoptotic cells in the MME-HVJ liposomes treated tumors compared with control tumors. MME was effectively expressed in the s.c. tumors treated with MME-HVJ liposomes, inducing angiostatin generation in those tumors, as demonstrated by Western blot analysis. In conclusion, our study demonstrated that repeated in vivo transduction of the MME gene directly into the tumors using HVJ-cationic liposomes suppressed the tumor growth by an antiangiogenic mechanism, providing, then, a feasible strategy for gene therapy of cancer. PMID- 11477588 TI - Antimetastatic and antitumor effects of benzoquinonoid AC7-1 from Ardisia crispa. AB - An antimetastatic and cytostatic substance, termed AC7-1, was isolated from Ardisia crispa and identified as a benzoquinonoid compound, 2-methoxy-6-tridecyl 1,4-benzoquinone. It was originally characterized as the potent PAF (platelet activating factor) receptor-binding antagonist with nonspecific antiplatelet effects on platelet aggregation induced by various agonists including PAF, ADP, thrombin and collagen. The nonspecific antiaggregatory properties of AC7-1 drew our interest given its possible relationship in integrin receptor-binding antagonistic activity. The integrin receptor plays an important role in metastasis and thrombosis as the cell surface transmembrane protein. Based on the aforementioned facts, the antimetastatic activities of AC7-1 were examined using various in vitro and in vivo metastasis assays. AC7-1 strongly blocked B16-F10 melanoma cell adhesion to extracellular matrix (ECM) and B16-F10 melanoma cell invasion. AC7-1 also remarkably inhibited pulmonary metastasis and tumor growth in vivo. AC7-1 inhibited B16-F10 melanoma cell adhesion to only specific synthetic peptides including RGDS. These findings suggest that antimetastatic activities of AC7-1 can be caused by blocking integrin-mediated adherence. We found AC7-1 to be a potential candidate for the development of a new antimetastatic drug. PMID- 11477589 TI - Incidence of leukemia and other cancers in Down syndrome subjects in Israel. AB - Epidemiologic data have confirmed the high susceptibility of persons with Down syndrome (DS) to leukemia. The question of proneness to other kinds of cancer is still open. In this study we reassessed the incidence rates of leukemia and other malignancies in Israeli DS subjects, based on the total population. The target population consisted of all DS subjects in Israel in the period of 1948--1995. Due to incompleteness of data, the target population was not fully achieved, thus the study population was divided into 2 subgroups: subjects born in Israel between 1979 and 1995 (registry group) and currently or past-institutionalized subjects born before 1979 (institution group). The cohort was linked with the Cancer Registry, and cancer cases that had been diagnosed through December 1995 were subsequently identified. Observed incidence rates were compared with expected rates in the general population. Standardized incidence ratios (SIR) and 95% confidence intervals (CI) were computed for each disease category. Analyses were performed separately for each subgroup of the study population. In the registry group, 7 cancer cases were observed, compared with 1.5 expected (SIR = 4.67, 95% CI 1.9--9.6), all leukemia cases. For the institution group a total of 17 cancer cases were observed, compared with 12.8 expected. These included 4 cases of leukemia (SIR = 6.90, 95% CI 1.90--17.70). An excess of gastric cancer in male subjects (SIR = 11.9, 95% CI 1.3--42.9) was also observed. Significant excess of leukemia in DS population in Israel is in accordance with previously published data. An excess of gastric cancer in DS male subjects born before 1979, which has not been reported before, should be further explored. PMID- 11477590 TI - Cohort study of thyroid cancer in a San Francisco Bay area population. AB - Using data from a large health plan, we performed a cohort study of thyroid cancer among 204,964 persons (aged 10--89 at baseline in 1964--1973, 54% female) followed for a median of 20 years. There were 196 incident thyroid cancers (73 in men, 123 in women). Risk was independently and positively related to female gender [relative risk (RR) = 1.56, 95% confidence interval (CI) = 1.12--2.19], Asian race (RR = 2.86, 95% CI = 1.76--4.65), completed college or post-graduate education (RR = 1.76, 95% CI = 1.20--2.59), history of goiter (RR = 3.36, 95% CI = 1.82--6.20), radiation of the neck region (RR = 2.33, 95% CI = 1.28--4.23) and family history of thyroid disease (RR = 2.18, 95% CI = 1.17--4.05). An inverse association was found for black race (RR = 0.55, 95% CI = 0.33--0.91). Cigarette smoking, alcohol consumption, personal history of hyperthyroidism, hypothyroidism, overweight or obesity, weight gain since age 20, height, occupational exposures, reproductive factors, oral contraceptives and hormone use did not show statistically significant relations to thyroid cancer. These results provide further evidence for a role of female gender, radiation, goiter, Asian race, high educational attainment and family history of thyroid disease in the etiology of thyroid cancer. PMID- 11477591 TI - Temporal changes in liver cancer incidence rates in Japan: accounting for death certificate inaccuracies and improving diagnostic techniques. AB - Primary liver cancer (PLC) rates have risen dramatically during the past few decades in some regions, particularly in Japan, where PLC is now the third major cause of cancer death. PLC is one of the most difficult tumors to diagnose correctly, because (i) the liver is a frequent site of cancer metastasis and (ii) death from PLC is often attributed to cirrhosis or chronic hepatitis. Also, because the disease is often rapidly fatal, a large proportion of liver cancer cases are identified based on death certificates alone without confirmation by clinical records. Thus, worldwide differences in published incidence rates for this disease reflect regional or national differences in both the accuracy of death certificates and the sensitivity of diagnostic methods. By comparing death certificate causes of death with those based on pathology review, we were able to adjust 1958--1994 incidence rates for a large Japanese cohort for these errors. Although the death certificate false-positive error rate declined, the false negative error rate remained high throughout the study. The introduction of improved liver cancer diagnostic methods in Japan in the early 1980s was associated with a sharp increase in PLC incidence. We conclude that errors in death certificate causes of death and changes in liver cancer diagnostic techniques have had an important impact on the reported incidence of this disease. Taking these factors into account, rates of hepatocellular carcinoma rose between 2.4- and 4.3-fold in our Japanese cohort from 1960 to 1985, peaked about 1993 and declined thereafter. Incidence rates of cholangiocarcinoma remained stable through 1987. PMID- 11477592 TI - Molecular forms of prostate-specific antigen in serum with concentrations of total prostate-specific antigen <4 microg/L: are they useful tools for early detection and screening of prostate cancer? AB - Molecular forms of prostate-specific antigen (PSA) improve the differentiation between benign prostatic hyperplasia (BPH) and prostate cancer (PCa) in men with total PSA concentrations between 4 and 10 microg/l. To evaluate the diagnostic utility of free PSA (fPSA) and complexed PSA forms for identification of men with PCa in the low PSA range of <4 microg/l, total PSA (tPSA), alpha(1) antichymotrypsin complexed PSA (PSA-ACT) and fPSA (Roche Elecsys [ES] system) as well as tPSA and complexed PSA (cPSA) (Bayer Immuno 1 system) were measured in archival serum samples from 31 untreated patients with PCa, 66 patients with BPH, and 90 men without prostatic disease. The median ratios of fPSA/tPSA, PSA ACT/tPSA and cPSA/tPSA were significantly different between patients with BPH and PCa (27.2 vs. 19.4%, 64 vs. 88%, 77.2 vs. 88.2%, p < 0.05). No associations between PSA forms and tumor stage and grade were found. Analysis of the receiver operating characteristic curves showed that these ratios could discriminate better between BPH and PCa patients than determination of the analytes tPSA, fPSA, cPSA and PSA-ACT alone. The use of one of the ratios would have eliminated roughly half of the unnecessary biopsies in this study. The ratios should be considered as potential tools to increase the selectivity of PCa detection at low PSA concentration. The ratios fPSA/tPSA and cPSA/tPSA can be determined using commercially available assays so that one of these ratios could be preferred instead of PSA-ACT determination. The ratios could be useful in assessing the risk of PCa in the individual and therefore in deciding on prostate biopsy for final diagnosis. PMID- 11477593 TI - A confocal study of spinal interneurons in living larval zebrafish. AB - We used confocal microscopy to examine the morphology of spinal interneurons in living larval zebrafish with the aim of providing a morphological foundation for generating functional hypotheses. Interneurons were retrogradely labeled by injections of fluorescent dextrans into the spinal cord, and the three dimensional morphology of living cells was reconstructed from confocal optical sections through the transparent fish. At least eight types of interneurons are present in the spinal cord of larval zebrafish; four of these are described here for the first time. The newly discovered cell types include classes of commissural neurons with axons that ascend, descend, and bifurcate in the contralateral spinal cord. Our reexamination of previously described cell types revealed functionally relevant features of their morphology, such as undescribed commissural axons, as well as the relationships between the trajectories of the axons of interneurons and the descending Mauthner axons. In addition to describing neurons, we surveyed their morphology at multiple positions along the spinal cord and found longitudinal changes in their distribution and sizes. For example, some cell types increase in size from rostral to caudal, whereas others decrease. Our observations lead to predictions of the roles of some of these interneurons in motor circuits. These predictions can be tested with the combination of functional imaging, single-cell ablation, and genetic approaches that make zebrafish a powerful model system for studying neuronal circuits. PMID- 11477594 TI - Borders and cytoarchitecture of the perirhinal and postrhinal cortices in the rat. AB - Cytoarchitectonic and histochemical analyses were carried out for perirhinal areas 35 and 36 and the postrhinal cortex, providing the first detailed cytoarchitectonic study of these regions in the rat brain. The rostral perirhinal border with insular cortex is at the extreme caudal limit of the claustrum, consistent with classical definitions of insular cortex dating back to Rose ([1928] J. Psychol. Neurol. 37:467-624). The border between the perirhinal and postrhinal cortices is at the caudal limit of the angular bundle, as previously proposed by Burwell et al. ([1995] Hippocampus 5:390-408). The ventral borders with entorhinal cortex are consistent with the Insausti et al. ([1997] Hippocampus 7:146-183) description of that region and the Dolorfo and Amaral ([1998] J. Comp. Neurol. 398:25-48) connectional findings. Regarding the remaining borders, both the perirhinal and postrhinal cortices encroach upon temporal cortical regions as defined by others (e.g., Zilles [1990] The cerebral cortex of the rat, p 77-112; Paxinos and Watson [1998] The rat brain in stereotaxic coordinates). Based on cytoarchitectonic and histochemical criteria, perirhinal areas 35 and 36 and the postrhinal cortex were further subdivided. Area 36 was parceled into three subregions, areas 36d, 36v, and 36p. Area 35 was parceled into two cytoarchitectonically distinctive subregions, areas 35d and 35v. The postrhinal cortex was divided into two subregions, areas PORd and PORv. These regional definitions of perirhinal areas 35 and 36 and the postrhinal cortex were confirmed by new empirical analyses of previously reported quantitative connectional data (Burwell and Amaral [1998a] J. Comp. Neurol. 398:179-205). PMID- 11477595 TI - Rhombomeric organization of vestibular pathways in larval frogs. AB - Rhombencephalic subnuclei and projection pathways related to vestibular function were mapped in larval ranid frogs. The retention of overt postembryonic rhombomeres (r) allowed direct visualization of the locations of neurons retrogradely labeled with fluorescent dextran amines from the midbrain oculomotor complex, cerebellum, vestibular nuclei, and spinal cord. Oculomotor projecting vestibular neurons were mainly located in bilateral r1/2, ipsilateral r3, and contralateral r5-8, and spinal projecting vestibular neurons mainly in ipsilateral r4 and contralateral r5. Vestibular commissural neurons were located in r1-3 and r5-7 and were largely excluded from r4. Cerebellar projecting neurons included contralateral inferior olivary neurons in r8 and vestibular neurons in bilateral r6/7 and contralateral r1/2. Mapping these results onto adult anuran vestibular organization indicates that the superior vestibular nucleus derives from larval r1/2, the lateral vestibular nucleus from r3/4, and the major portions of the medial and descending vestibular nuclei from r5-8. The lateral vestibulospinal tract projects from an origin in r4, whereas a possible ascending tract of Deiters arises in r3. Rhombomere 5 contains a nuclear group that appears homologous to the tangential nucleus of fish, reptiles, and birds and thus likely serves gravistatic and linear vestibulomotor reflexes. Comparisons between frogs and other vertebrates suggest that vestibular neurons performing similar computational roles during head movements originate from the same segmental locations in different species. PMID- 11477596 TI - Quantitative analysis of the dendrites of sacral preganglionic neurons in the cat. AB - Quantitative analyses were performed on the dendrites and somata of 25 electrophysiologically identified preganglionic neurons (PGN) obtained from the sacral spinal cord of the cat by intracellular injection of Neurobiotin or horseradish peroxidase. Total dendritic length and surface area were measured for each dendrite. The sizes of the stem dendrites measured at their base were positively correlated with the sizes of the entire tree and numbers of end branches. Total surface area of somata and dendrites averaged 39,138 microm(2); 90.7% of that was from the dendrites. To obtain measurements of the relative contributions of PGN dendrites to specific regions of the spinal cord, the percentage of each dendrite occupying eight spinal cord regions was recorded. Sixty-three percent of the dendrites projected dorsal to their somata, whereas an average of 33.3% of dendrites were located in the white matter, most of them in the lateral and dorsolateral funiculi. The neurons within this sample formed a continuum with some neurons having a large percentage of dendrites in lamina I but little in the white matter, whereas at the other end of the continuum were cells with the reverse configuration. The intermediate neurons had dendrites in both locations. Taken together, these data indicate a heterogeneous population of PGN in the lateral band of the sacral parasympathetic nucleus. PMID- 11477597 TI - Identifiable nitrergic neurons in the central nervous system of the nudibranch Melibe leonina localized with NADPH-diaphorase histochemistry and nitric oxide synthase immunoreactivity. AB - Nitric oxide (NO) is a gaseous intercellular messenger produced by the enzyme nitric oxide synthase (NOS). In this study, we used two different techniques nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and NOS immunocytochemistry-to demonstrate that NOS is present in a pair of identifiable cells in the central nervous system of the nudibranch Melibe leonina. In the Melibe brain, NADPH-d histochemistry revealed only a single pair of bilaterally symmetrical cells in the cerebropleural ganglia. NOS activity also was found in the neuropil of the cerebral, pedal, and buccal ganglia; in the tentacles of the oral hood; in the sensory end of the rhinophores; and in the epithelial tissue of the mouth, preputium, and glans penis. Immunocytochemistry using NOS antisera corroborated the results of the NADPH-d histochemistry by staining the same two cells in the cerebropleural ganglia. Each of these identifiable nitrergic neurons projects into the ipsilateral pedal ganglion. Because the pedal ganglia play a critical role in the control of locomotion, our results provide morphological evidence suggesting that NO may influence swimming or crawling in Melibe leonina. PMID- 11477598 TI - Neuromodulator content of hamster intergeniculate leaflet neurons and their projection to the suprachiasmatic nucleus or visual midbrain. AB - The intergeniculate leaflet (IGL) of the lateral geniculate complex has widespread, bilateral, and reciprocal connections with nuclei in the subcortical visual shell. Its function is poorly understood with respect to its role in visual processing. The most well-known IGL projection, and the only one with a clear function, is the geniculohypothalamic tract (GHT) that terminates in the suprachiasmatic nucleus (SCN), site of the primary circadian clock. The hamster GHT is derived, in part, from IGL neurons containing neuropeptide Y and enkephalin. IGL neurons containing these peptides also project to the pretectal region. The present studies used a combination of immunohistochemical, lesion, and retrograde tracing techniques to study neuron types in the IGL and their projections to hamster SCN and pretectum. Two additional neuromodulators, gamma aminobutyric acid (GABA) and neurotensin, are shown to be present in IGL neurons. The GABA- and neurotensin-immunoreactive neurons project to the SCN with terminal field patterns very similar to those for neuropeptide Y and enkephalin. IGL neurons of all four types also send projections to the pretectum, but rarely do individual cells project to both the SCN and the pretectum. Nearly all neurotensin is colocalized with neuropeptide Y in IGL neurons, although about half of the neuropeptide Y cells do not contain neurotensin. Otherwise, the extent to which the four neuromodulators are colocalized varies from 6% to 54%. Nearly every SCN neuron appears to contain GABA. In the IGL, the majority of cells studied are not identifiable by GABA immunoreactivity. Putative functions of the various neuromodulator projections from the IGL to pretectum or SCN are discussed. PMID- 11477599 TI - Localization and quantification of 5-hydroxytryptophan and serotonin in the central nervous systems of Tritonia and Aplysia. AB - Serotonin (5-hydroxytryptamine, 5-HT) plays a central role in several behaviors in marine molluscs and other species. In an effort to better understand the regulation of 5-HT synthesis, we used high performance liquid chromatography (HPLC) with electrochemical detection and immunohistochemistry to measure and map the distribution of the immediate precursor of 5-HT, 5-hydroxytryptophan (5-HTP), in two model opisthobranch molluscs, the nudibranch Tritonia diomedea and the anaspid Aplysia californica. HPLC measurements showed that 5-HTP is present at approximately the same level as the 5-HT metabolite, 5-hydroxyindolacetic acid (5 HIAA) but is more than 100 times lower in concentration than either 5-HT or dopamine in the same tissue. Specific 5-HTP immunoreactivity was colocalized with serotonin in both species. The overall intensity of 5-HTP immunoreactivity in individual ganglia agreed with HPLC measurements for those ganglia. The intensity of 5-HTP immunolabeling varied between cell types and was correlated with the intensity of 5-HT immunolabeling. In particular, differences in staining intensity were consistently seen among the three dorsal swim interneurons of the Tritonia swim central pattern generator circuit. Some nonserotonergic neurons also displayed low levels of 5-HTP immunolabeling that were above background levels. Together, these results support the notion that production of 5-HTP is a rate-limiting step in serotonin synthesis and suggest that there may be additional regulation that allows 5-HTP to accumulate to varying levels. PMID- 11477600 TI - Nimodipine promotes regeneration and functional recovery after intracranial facial nerve crush. AB - The calcium flow inhibitor, nimodipine, has been shown to promote motor neuron survival in the facial nucleus after intracranial facial nerve transection. However, it has not been known whether the neuroprotective effects primarily involve survival of nerve cell bodies or outgrowth and/or myelination of nerve fibers. Here, we studied the effects of nimodipine in a different injury model in which the facial nerve was unilaterally crushed intracranially. This lesion caused complete anterograde degeneration and partial retrograde degeneration that were studied with a combination of several stereological methods. Nimodipine did not attenuate the modest lesion-induced neuronal loss (13%) but accelerated the time course of functional recovery and axonal regrowth, inducing increased numbers and sizes of myelinated axons in the facial nerve. It is interesting to note that nimodipine also enlarged the axons and the myelin sheaths in the nonlesioned facial nerve, which points to the possibility of using this substance for new clinical applications to promote axonal growth and remyelination. PMID- 11477601 TI - Stereologic description of the changing expression of constitutive nitric oxide synthase and heme oxygenase in the enteric plexuses of the pig small intestine during development. AB - The similarities between heme oxygenase-2 (HO-2) and nitric oxide synthase (nNOS) and the transient expression of nNOS during development led us to investigate whether both systems are similarly affected by changes that occur during development and by regional differences along the small intestine. By combining NADPH diaphorase histochemistry and HO-2 immunohistochemistry on whole-mount preparations and by using stereologic methods, a qualitative and quantitative description of HO-2 and nNOS expression was obtained. Examinations were carried out on the small intestine of fetal, 1-2-day and 5-6-week-old pigs. In all age groups, three enteric plexuses were distinguished. The presence of HO-2 immunoreactive (HO-2-IR) and NADPH diaphorase-positive neurons corresponded to earlier morphological and physiological reports. Nevertheless, the total number of nitrergic neurons remained constant or decreased in the enteric plexuses, whereas the total number of HO-2-IR neurons displayed an overall increase. Changing concentrations of glucocorticoids, target-derived signals, presynaptic input, and an effect of HO-2 activity on nNOS synthesis are likely to play roles in the observed developmental changes. The numerical density of HO-2-IR neurons remained relatively constant along the intestinal tract; in contrast, the nitrergic neurons were most numerous in the inner submucous and myenteric plexus in the duodenum and ileum, respectively. It is believed that the duodenal nitrergic neurons in the inner submucous plexus could be involved in the regulation of duodenal secretion processes, whereas the region-dependent density in the myenteric plexus possibly forms the morphological basis for a regionally different participation of NO in the relaxation of the small intestine. PMID- 11477602 TI - Meier-Gorlin syndrome: report of eight additional cases and review. AB - The Meier-Gorlin syndrome or ear, patella, short stature syndrome (MIM 224690) is a rare autosomal recessive disorder, characterized by the association of bilateral microtia, aplasia/hypoplasia of the patellae, and severe pre- and postnatal growth retardation. Twenty-one cases have been reported in literature thus far. Here we report on eight patients from seven families and compare them with previously described cases. One of the present cases had previously undescribed genital anomalies. There is a difference in facial characteristics between patients reported in early infancy and those described at older age; follow-up of patients is needed to substantiate this changing facial phenotype. We recommend radiographic survey of the patellae in patients at older age to investigate the weight of absent or hypoplastic patellae in the diagnosis of the syndrome. PMID- 11477603 TI - Cohen syndrome: essential features, natural history, and heterogeneity. AB - This article elucidates the clinical picture in Cohen syndrome (MIM 216550), an autosomal recessive disorder that is overrepresented in Finland. The diagnosis is based on the typical clinical picture: nonprogressive psychomotor retardation, motor clumsiness and microcephaly, typical facial features, childhood hypotonia and hyperextensibility of the joints, ophthalmologic findings of retinochoroidal dystrophy and myopia in patients over 5 years of age, and granulocytopenia. In a nationwide study, 29 Finnish patients were investigated. Magnetic resonance images of the brain with quantitative structure analyses revealed a relatively enlarged corpus callosum (CC). The youngest patients had normal EEGs, while all others had low-voltage EEGs. Of the patients, 22% had profound, 61% severe, 6% moderate, and 11% mild retardation. In an adaptive behavior scale (AAMD), patients had high scores in the positive domains (self-direction, responsibility, and socialization), whereas maladaptive behavior was almost lacking. Only the youngest patients had unimpaired visual function. Vision started to deteriorate early but slowly. Progressive myopia and retinochoroidal dystrophy were found in all of the patients over 5 years of age. All of the patients had isolated granulocytopenia. The heart anatomy was normal. However, decreased left ventricular function with advancing age was found. No significant endocrine abnormalities were discovered. Fingers were slender but short, with a typical metacarpophalangeal pattern profile. The manifestations vary at different ages. The Finnish Cohen patients are clinically highly homogeneous, their disease gene being located on chromosome 8. Heterogeneity probably exists among other patients claimed to have Cohen syndrome. PMID- 11477604 TI - Successful pregnancy in a woman with Bloom syndrome. AB - Bloom syndrome is a rare autosomal recessive disorder notable for increased chromosome fragility and an increased rate of somatic mutation. The clinical manifestations include small stature, a characteristic dermatologic lesion, and an excess incidence of malignancy. Fertility is generally reduced. A 19-year-old white woman with Bloom syndrome was successfully treated for preterm labor at 32 weeks' gestation, and ultimately delivered a healthy male infant at 35 weeks' gestation. Reports of pregnancy in women with Bloom syndrome are few. Despite reduced fertility, conception can occur, and women with Bloom syndrome should receive appropriate reproductive counseling to prevent unintended pregnancies and increased surveillance for preterm birth. PMID- 11477605 TI - Trisomy 4 pter-q12 and monosomy of chromosome 13 pter-q12 in a male with deficiency of all blood lymphocyte populations. AB - A six-year-old male presented with multiple congenital anomalies, mental retardation, developmental delay, and an increased frequency of upper and lower respiratory infections and deficiency of all blood lymphocyte populations. Chromosome analysis showed an unbalanced translocation involving chromosomes 4 and 13, leading to partial trisomy for 4pter-q12 and partial monosomy for 13pter q13 [karyotype, 46,XY,+der(4)t(4;13)(q12;q12),-13)]. The mother is the carrier of a balanced translocation involving chromosomes 4 and 13. The translocation is known to be segregating for three generations in this family. The child was found to have deficiency of all blood lymphocyte populations, but other hemopoietic lineages appeared to be normal. In addition, his fresh T cells were principally CD45RA+, CD62L+, and deficient in the Fas receptor. This deficiency of all blood lymphocyte populations may be due to an overdose of a gene or genes located in the region of chromosome 4 or a partial deficiency of a gene or genes in the region of chromosome 13 that regulate the development of the lymphocyte lineage. Since the mother contributed two copies of chromosomal region 4pter-q12 and no copy of 13pter-q12, the deficiency of all blood lymphocyte populations in our patient may be the result of either uniparental disomy or imprinting. A maternal granduncle with dissimilar dysmorphic features was not lymphopenic but was neutropenic. PMID- 11477606 TI - Carrier identification by FISH analysis in isolated cases of X-linked ichthyosis. AB - X-linked ichthyosis (XLI) is an inborn error of metabolism due to steroid sulfatase (STS) deficiency. STS assay and FISH are useful in diagnosing carrier status of XLI. Biochemical analysis appears to indicate that most sporadic cases are inherited. Since this method does not seem to be completely reliable in recognizing XLI-carriers, the aim of the present study was to corroborate by FISH whether or not most sporadic cases of XLI had de novo mutations. XLI patients were classified through STS assay and PCR amplification of 5'-3' ends of the STS gene. XLI patients had undetectable levels of STS activity and complete deletion of the STS gene. Patients' mothers were studied through STS assay and FISH. Nine out of 12 mothers presented an STS activity compatible with XLI-carrier state. These mothers also had only one copy of the STS gene, indicating that they carry the primary gene defect. One mother had normal STS activity but only one copy of the STS gene. This data corroborated that most sporadic cases do not represent de novo mutations, and that FISH must be included in the analysis of mothers of sporadic cases when they present with normal STS activity, in order to correctly diagnose the XLI carrier state. PMID- 11477607 TI - Association of arrhythmia and sudden death in macrocephaly-cutis marmorata telangiectatica congenita syndrome. AB - Macrocephaly-cutis marmorata telangiectatica congenita (M-CMTC) constitutes a distinct entity characterized by prenatal overgrowth, macrosomia, hemihypertrophy, macrocephaly, nonobstructive hydrocephaly, frontal bossing, hypotonia, developmental delay, generalized or facial capillary malformation with upper philtral nevus flammeus and cutis marmorata, joint hypermobility, loose skin, toe syndactyly, and postaxial polydactyly. All but one of the cases reported previously had benign clinical courses without showing an increased risk of early infant death. We describe three additional cases with poor clinical outcomes including severe postnatal growth failure, intractable cardiac arrhythmia in two cases, and sudden infant death in two cases. Arrhythmia has not been described previously as one of the symptoms of M-CMCT. Patients with M-CMTC associated with severe postnatal growth failure and arrhythmia may constitute a distinct clinical subtype of M-CMTC with an increased risk of life-threatening episodes or sudden death. Recognizing this clinical subtype of M-CMTC is important to prevent these serious potential complications. PMID- 11477608 TI - Severe mental retardation, short stature, facial anomalies, joint laxity, and dislocations in two sisters: previously undescribed MCA/MR syndrome. AB - We describe two sisters with short stature, obesity, "bulbous" nasal tip, microretrognathism, brachydactyly, joint hyperlaxity and dislocation, and mental retardation. Skeletal surveys disclosed widened mandibular angles, thin temporal processes, hypoplastic clavicles, short distal ends of ulnae, short fourth metacarpals, and dislocation of hips, elbows, and thumbs. The parents are first cousins. To the best of our knowledge, this combination of multiple congenital anomalies and mental retardation has not been reported before. PMID- 11477609 TI - Sex chromosome mosaicism in gonads of a fetus with cystic hygroma and deletion of the short arm of Y chromosome including loss of SRY. AB - The SRY gene on the short arm of the Y chromosome is necessary for male development. Without SRY, patients with 46,XY karyotype develop as females, fail to achieve normal puberty and have dysgenic gonads and a high incidence of gonadoblastoma. Here we report a female fetus, aborted at 17 weeks of pregnancy, with a non-mosaic 46,X,del(Y)(p11.2).ish del(Y)(SRY-) karyotype diagnosed by classical cytogenetics and fluorescence in situ hybridization (FISH). Ovarian tissue was full of oocytes and mitotic figures. FISH studies of ovarian tissues with X and Y centromere probes revealed extensive sex chromosome mosaicism, manifested by loss of the Y chromosome and polysomy of the X chromosome. We propose that X chromosome polysomy is a post-zygotic event that arises to facilitate gonadal differentiation in the absence of all factors necessary for normal gonadal development. PMID- 11477610 TI - GPC3 mutation analysis in a spectrum of patients with overgrowth expands the phenotype of Simpson-Golabi-Behmel syndrome. AB - Simpson-Golabi-Behmel syndrome (SGBS) is an X-linked overgrowth syndrome caused by deletions in glypican 3 (GPC3). SGBS is characterized by pre- and postnatal overgrowth, a characteristic facial appearance, and a spectrum of congenital malformations which overlaps that of other overgrowth syndromes. We performed GPC3 deletion screening on 80 male patients with somatic overgrowth in the following categories: SGBS (n = 19), possible SGBS (n = 26), including families in which individuals had previously been diagnosed with other overgrowth syndromes, and Wiedemann-Beckwith syndrome (WBS) (n = 35). Using exon-specific PCR and Southern blot analysis, we identified seven GPC3 deletions. In most cases a clear X-linked family history was not present. In two cases, GPC3 deletions were identified in patients belonging to pedigrees published previously as other overgrowth syndromes: one with a diagnosis of Sotos syndrome and the other Perlman syndrome with nephroblastomatosis. A third patient developed hepatoblastoma, a tumor type not previously described in SGBS. No GPC3 deletions were identified among the WBS patients. Direct sequencing of all GPC3 exons in the remaining 13 SGBS patients without GPC3 deletions did not identify any further mutations, raising the possibility of alternative silencing mechanisms and/or other genes in the pathogenesis of SGBS. Our results validate the clinical specificity of the facial appearance, skeletal/hand anomalies, and supernumerary nipples in patients with GPC3 deletions. Our data also suggest that nephroblastomatosis and hepatoblastoma are included in the phenotypic spectrum of GPC3 deletions and SGBS, underscoring the importance of tumor surveillance in these children. PMID- 11477611 TI - Maternal isochromosome 7q and paternal isochromosome 7p in a boy with growth retardation. AB - A 12-year 9-month-old boy with postnatal growth retardation, normal psychomotor development, and minor anomalies that included a triangular-shaped face, small nose, and narrow and high-arched palate is reported. The constitutional karyotype was 46,XY,i(7)(p10),i(7)(q10). Molecular investigations revealed the presence of a maternal isodisomy 7q and a paternal isodisomy 7p. The clinical and molecular findings are notably congruent with a recently reported case and support the hypothesis of one or more maternally imprinted genes located on the long arm of chromosomes 7 that regulate, in particular, postnatal growth. PMID- 11477612 TI - Defining the breakpoints of proximal chromosome 14q rearrangements in nine patients using flow-sorted chromosomes. AB - The breakpoints of deletions and translocations in the proximal chromosome 14q region were defined in nine patients, four of whom have not been reported previously. The aberrant chromosomes were isolated by flow cytometry and used to map the chromosome 14 deletion or translocation breakpoints. The parental origins of deletions were ascertained as paternal in five cases and maternal in one. With the draft genomic sequence for human chromosome 14 available, gene searches were performed on selected intervals of the 14q11.2-q21 region to identify candidate genes for the observed phenotype in some of those affected. Gain of function of the gene PAX9 on chromosome 14 is a possible candidate for a t(14;18) patient affected with mesomelic bone dysplasia. Furthermore, a compilation of other human chromosome 14q proximal deletion and translocation cases was obtained from a search on cytogenetic databases. These findings suggest a locus for myelofibrosis at chromosome 14q13. This study contributes to useful information for identifying disease genes in this region. PMID- 11477613 TI - Antenatal presentation of carnitine palmitoyltransferase II deficiency. AB - Carnitine palmitoyl transferase (CPT) II deficiency is usually manifested around puberty by exercise induced myoglobinuria. Two Ashkenazi Jewish sibs with the rare antenatal form of CPTII deficiency are reported. On the 5th gestational month periventricular calcifications and markedly enlarged kidneys were found in both of them. The activity of CPTII in lymphocytes was undetectable and both sibs were homozygous for the 1237delAG mutation. Because of the serious consequences of homozygosity for this mutation, genotype determination of all Ashkenazi patients with the adolescent form of CPTII deficiency is warranted. PMID- 11477614 TI - Two HLA DRB 1 alleles confer independent genetic susceptibility to Graves disease: relevance of cross-population studies. AB - Recent studies of Graves disease (GD) employing genome scanning techniques excluded the major histocompatibility complex as a contributor to disease liability. These findings contradict earlier population association studies. Our own earlier studies have also emphasized that genetic variation in human populations may give novel clues to disease liability and manifestations. To this end, we studied HLA class II alleles in 47 Latvian GD patients and 111 matched healthy controls. As expected, we found that DRB1*03 and DQA1*0501 (OR = 3.6, P = 0.029 and OR 2.35, P = 0.0373, respectively) were associated with GD. Unforeseen, DRB1*04 was found to be significantly increased in the patients compared to controls (OR 3.267, corrected P = 0.0319). The two DRB1 alleles conferred two non overlapping and independent susceptibilities to GD, in that only three patients were positive for both alleles, and the removal of each allele in turn resulted in only the other DRB1 allele showing significant association with the disease. There was no heterogeneity between the two patient groups (DRB1*03 positive and DRB1*04 positive) in clinical characteristics or disease manifestations. The phenotype DRB1*03 and/or DRB1*04 was found in 34/47 patients compared to 27/111 controls yielding an OR of 7.395 (P corrected = 0.000019). We examined the structural basis of DRB1 susceptibility to GD in light of this and previous studies, showing that DRB1*03, 04, and 08 were positively associated with the disease, whereas DRB1*07 was negatively associated. Differences in protein sequences were noted at residues 54, 57, 59, and 66; positions 54, 57, and 66 are on the same face of the alpha helix. The canonical arginine 54 is replaced by glutamine in DRB1*07. At position 66, asparagine in DRB1*03 and tyrosine in DRB1*04 are replaced by phenylalanine in DRB1*07. Residue 59, likely involved in pocket formation in the antigen binding groove, is modified by replacement of tyrosine in DRB1*03, 08, and 04 and by leucine in DRB1*07. The predicted differences in the shape and charges of the proximal reaches of the antigen binding groove between DRB1*07, and 03, 04, and 08, could determine whether or not a peptide from an auto-antigen would be bound or not. Genetic variation among human populations may yield important clues to specific disease liability. PMID- 11477615 TI - Three cell line mosaicism involving structural and numerical abnormalities of chromosome 18 in a 3.5-year-old girl: 47,XX,+18/47,XX,+del(18)(q22)/46,XX. AB - We report on a 3.5-year-old girl with a mosaic karyotype including full trisomy 18, normal cells and a majority of cells with partial trisomy involving an extra chromosome 18 deleted at band q22. She had cardiac and CNS anomalies, dysmorphic facial features failure to thrive and developmental delay. A gastrostomy tube was placed at 2 years of age. The combination of improved nutrition and optimal developmental therapy has led to her sitting supported, attempting to stand and enhancement of her cognitive and non-verbal communication abilities. Molecular investigation of the patient and her parents using microsatellite analysis has led to the conclusion that, as expected, the additional copy of chromosome 18 constituting the full trisomic cell line is maternal meiosis I in origin. The data, however, indicate that in the trisomic cell line containing the deleted chromosome 18q, the structurally abnormal 18 was of paternal origin. We think this case is the first described with both structural and numerical trisomic mosaicism involving chromosome 18 in a liveborn infant. We propose a mechanism of origin and review the literature, comparing the clinical presentation of this case with individuals having full or partial trisomy 18. PMID- 11477616 TI - Localization of non-specific X-linked mental retardation gene (MRX73) to Xp22.2. AB - Clinical and molecular studies are reported on a family (MRX73) of five males with non-specific X-linked mental retardation (XLMR). A total of 33 microsatellite and RFLP markers was typed. The gene for this XLMR condition was been linked to DXS1195, with a lod score of 2.36 at theta = 0. The haplotype and multipoint linkage analyses suggest localization of the MRX73 locus to an interval of 2 cM defined by markers DXS8019 and DXS365, in Xp22.2. This interval contains the gene of Coffin-Lowry syndrome (RSK2), where a missense mutation has been associated with a form of non-specific mental retardation. Therefore, a search for RSK2 mutations was performed in the MRX73 family, but no causal mutation was found. We hypothesize that another unidentified XLMR gene is located near RSK2. PMID- 11477617 TI - Down syndrome before Down: a retrospect. AB - We have attempted to identify published descriptions and reports of skeletal remains, figurines and paintings of suggested or supposed instances of Down syndrome predating John Langdon Haydon Down's initial account of the condition in 1866. A review of the ascertained material in each of these categories has not provided, in our opinion, convincing evidence of presence of the syndrome, although that possibility is not excluded. In the interests of historical authenticity readers are invited to point out perceived errors or omissions in this presentation. PMID- 11477618 TI - Nablus mask-like facial syndrome. PMID- 11477620 TI - Whistling face syndrome with normal hands and feet. PMID- 11477621 TI - Comments on "gradient-induced acoustic and magnetic field fluctuations in a 4T whole-body MR imager". AB - The results published in the article Gradient-Induced Acoustic and Magnetic Field Fluctuations in a 4T Whole-Body MR Imager by Wu et al. (Magn Reson Med 2000;44:532-536) appear to be consistent with the response of a time-stationary linear system. Since a linear system is more simply described than a nonlinear system, the authors are urged to reanalyze their data to test the linear-system hypothesis. PMID- 11477622 TI - Xenon-129 MR imaging and spectroscopy of rat brain using arterial delivery of hyperpolarized xenon in a lipid emulsion. AB - Hyperpolarized (129)Xe dissolved in a lipid emulsion constitutes an NMR tracer that can be injected into the blood stream, enabling blood-flow measurement and perfusion imaging. A small volume (0.15 ml) of this tracer was injected in 1.5 s in rat carotid and (129)Xe MR spectra and images were acquired at 2.35 T to evaluate the potential of this approach for cerebral studies. Xenon spectra consistently showed two resonances, at 194.5 ppm and 199.0 ppm relative to the gas peak. The signal-to-noise ratio (SNR) obtained for the two peaks was sufficient (ranging from 12 to 90) to follow their time courses. 2D transverse projection xenon images were obtained with an in-plane resolution of 900 microm per pixel (SNR range 8-15). Histological analysis revealed no brain damage except in two rats that had received three injections. PMID- 11477623 TI - High-resolution (1)H NMR spectroscopy in organs and tissues using slow magic angle spinning. AB - It is shown that high-resolution (1)H NMR spectra of intact excised tissues and organs can be obtained by rotating the sample slowly about an axis at the magic angle of 54 degrees 44' with the external magnetic field. In this way tissue and cellular damage invoked by standard magic angle spinning (MAS) experiments, where spinning speeds of several kHz are typically employed, are minimized. Special RF pulse sequences, developed originally in solid state NMR, can be used to produce a spinning sideband-free isotropic spectrum. In this article the first results are shown of the brain, heart, liver, gluteus muscle, and kidney excised from mice using the 2D-phase-altered spinning sidebands (PASS) technique and employing MAS spinning speeds of 43-125 Hz. It was found that with slow sample spinning similar, and in some cases even better, spectral resolutions are obtained as compared with fast MAS. PMID- 11477624 TI - Volumetric multishot echo-planar spectroscopic imaging. AB - Rapid volumetric magnetic resonance spectroscopic imaging (MRSI) is potentially of great relevance to the diagnosis and treatment of focal cerebral diseases such as cancer and epilepsy. A strategy for volumetric multishot echo-planar spectroscopic imaging (MEPSI) is described which allows whole-brain metabolite mapping in approximately 20 min. A multishot trajectory is used in both the spatial and temporal domains which reduces the accumulated phase during each echo train and tolerates conventional Fourier reconstruction without regridding. Also described is a generalized correction for phase discontinuities arising from the multishot acquisition of the time domain, which is independent of the spatial k space trajectory and is therefore also applicable to multishot spiral MRSI. Whole brain, lipid-suppressed MEPSI data were acquired from five normal subjects. The mean signal-to-noise ratios (SNRs) (+/-SE) for the n-acetylaspartate (NAA), choline (Cho), and creatine (Cr) maps across all subjects were 21.3 +/- 1.8, 11.7 +/- 0.6, and 9.2 +/- 0.6, respectively, with a computed voxel size of 2.33 ml. PMID- 11477625 TI - Analysis of volume MRI and MR spectroscopic imaging data for the evaluation of patients with brain tumors. AB - The combination of volumetric MRI and multivoxel localized MR spectroscopic imaging (MRSI) data provides the potential for quantifying variations in tissue morphology and function. These techniques have numerous applications for the evaluation of neurological diseases. While state-of-the-art whole-body MR scanners are able to acquire both types of data, there have been relatively few reports which have presented clinical applications of the technology. One of the reasons for this has been the need to develop software for reconstruction and reliable interpretation of 3D imaging and spectral data. In this article, we describe the strategy developed for quantitative analysis of combined MRI and MRSI examinations from patients with brain tumors and evaluate the key components of this procedure using both simulations and empirical data from phantoms, normal volunteers, and patients. Important factors are the use of volume or interleaved multislice anatomic images as a reference, the reconstruction and correction of the spectral data for frequency, phase, and baseline distortions and consideration of the characteristics of the coil, RF pulses used for spatial selection, and phase encoding procedures. These studies show that the metabolic parameters most critical for distinguishing tumor from normal and necrotic tissue were relative levels of choline and NAA. Levels of creatine and lactate were found to be variable, both in terms of their spatial distribution within individual lesions and between different patients. PMID- 11477626 TI - Quantitative (1)H magnetic resonance spectroscopy of myoglobin de- and reoxygenation in skeletal muscle: reproducibility and effects of location and disease. AB - 1H-magnetic resonance spectroscopy ((1)H-MRS) of deoxymyoglobin (DMb) provides a means to noninvasively monitor the oxygenation state of human skeletal muscle in work and disease. As shown in this work, it also offers the opportunity to measure the absolute tissue content of DMb, the basic oxygen consumption of resting muscle, and the reperfusion characteristics after release of a pressure cuff. The methodology to determine these tissue properties simultaneously at two positions along the calf is presented. The obtained values are in agreement with invasive determinations. The reproducibility of the (1)H-MRS measurements is established for healthy controls and patients with peripheral arterial disease (PAD). A location dependence in axial direction, as well as differences between controls and patients are demonstrated for all parameters. The reoxygenation time in particular is expected to provide a means to quantitatively monitor therapies aimed at improving muscular perfusion in these patients. PMID- 11477627 TI - Abnormal lipid profile of dystrophic cardiac tissue as demonstrated by one- and two-dimensional magic-angle spinning (1)H NMR spectroscopy. AB - Dystrophin, a protein associated with sarcolemma and cell membranes, is not expressed in sufferers of Duchenne muscular dystrophy (DMD), or in the mdx mouse. DMD is a fatal disorder, with a significant proportion of fatalities associated with cardiac failure ( approximately 40% having dilated cardiomyopathy and >90% clinically significant cardiac defects at death). In this study, the metabolic composition of intact dystrophic cardiac tissue was investigated using high resolution magic-angle spinning (HRMAS) (1)H NMR spectroscopy with both 1- and 2D pulse sequences coupled with pattern recognition (PR). While conventional solvent presaturation spectra indicated increases in CH(2) chain length in lipids, PR analysis of correlation spectroscopy (COSY) spectra demonstrated that this was also accompanied by an increase in concentration of lactate or threonine along with a relative decrease in CH = CHCH(2)CO groups in these lipids. To investigate the physical environment of these lipids, T(2)- and diffusion-weighted (1)H MAS NMR spectra were acquired on whole-tissue samples. The relatively increased lipid signal intensity in dystrophic tissue was due to an increase in molecules with long T(2) and short diffusion rates. The use of a range of pulse programs allowed the direct probing of the biochemical environment in which the lipid infiltration occurred, and by coupling the experiments to PR the significance of lipid infiltration and accumulation was also assessed. PMID- 11477628 TI - Volume correction for edema in single-volume proton MR spectroscopy of contrast enhancing multiple sclerosis lesions. AB - The effect of edema on metabolic changes in contrast-enhancing multiple sclerosis lesions was studied by combining quantification of proton MR spectra with segmentation of the volume-of-interest, which was based on biexponential T(2) relaxation. All lesions showed a second component (s(long)) with a longer T(2) (185-450 ms), which was increased compared to healthy controls. Regression analysis indicated that s(long) replaces the short-T(2) component and total creatine. Since the water content was close to 100%, s(long) was used to correct for an increase in extracellular space. This compensated for the apparent loss of creatine and rendered cholines markedly increased, as observed in animals with experimental allergic encephalomyelitis. Total N-acetyl aspartate (NAA) concentration was inversely correlated with s(long) and between 34-70% of its average reduction was assigned to edema. Thus, NAA loss exceeded cellular loss. Assessment of varying degrees of edema may be especially beneficial for quantitative longitudinal studies. PMID- 11477629 TI - Effect of graded hypo- and hypercapnia on fMRI contrast in visual cortex: quantification of T(*)(2) changes by multiecho EPI. AB - The sensitivity of functional magnetic resonance imaging (fMRI) in visual cortex to graded hypo- and hypercapnia was quantified in 10 normal subjects using single shot multiecho echo-planar imaging (Turbo-PEPSI) with eight equidistant echo times (TEs) between 12 and 140 ms. Visual stimulation was combined with controlled hyperventilation and carbon dioxide inhalation to perform fMRI at six levels of end-expiratory pCO(2) (PETCO(2)) between 20 and 70 mm Hg. T(*)(2) in visual cortex during baseline conditions (light off) increased nonlinearly from 20 to 70 mm Hg, from 61.1 +/- 4.2 ms to 72.0 +/- 4.6 ms. Changes in T(*)(2) due to visual stimulation increased 2.1-fold, from 1.2 +/- 0.6 ms at 20 mm Hg to 2.5 +/- 0.7 ms at 50 mm Hg. An almost complete loss of functional contrast was measured at 70 mm Hg. The model of MR signal dephasing by Yablonskiy and Haacke (Mag Reson Med 1994;32:749-763) was used to predict changes in cerebral blood flow (CBF), which were found to be consistent with results from previous positron emission tomography (PET) studies. This study further emphasizes that global CBF changes (due to PETCO(2) changes even in the physiological range) strongly influence fMRI contrast and need to be controlled for. PMID- 11477630 TI - Quantification of the effect of water exchange in dynamic contrast MRI perfusion measurements in the brain and heart. AB - Measurement of myocardial and brain perfusion when using exogenous contrast agents (CAs) such as gadolinium-DTPA (Gd-DTPA) and MRI is affected by the diffusion of water between compartments. This water exchange may have an impact on signal enhancement, or, equivalently, on the longitudinal relaxation rate, and could therefore cause a systematic error in the calculation of perfusion (F) or the perfusion-related parameter, the unidirectional influx constant over the capillary membranes (K(i)). The aim of this study was to quantify the effect of water exchange on estimated perfusion (F or K(i)) by using a realistic simulation. These results were verified by in vivo studies of the heart and brain in humans. The conclusion is that water exchange between the vascular and extravascular extracellular space has no effect on K(i) estimation in the myocardium when a normal dose of Gd-DTPA is used. Water exchange can have a significant effect on perfusion estimation (F) in the brain when using Gd-DTPA, where it acts as an intravascular contrast agent. PMID- 11477631 TI - Measurement of tissue oxygen extraction ratios from venous blood T(2): increased precision and validation of principle. AB - It has recently been shown that parenchymal oxygen extraction ratios (OERs) can be quantified using the absolute T(2) of venous blood draining from this tissue (Oja et al., J Cereb Blood Flow Metab 1999;19:1289-1295). Here, a modified Carr Purcell-Meiboom-Gill (CPMG) multiecho experiment was used to increase the efficiency and precision of this approach and to test the applicability of the two-compartment exchange model for spin-echo BOLD effects in pure venous blood. Relaxation measurements on bovine blood as a function of CPMG interecho spacing, oxygen saturation, and hematocrit provided the baseline relaxation and susceptibility shift parameters necessary to directly relate OER to T(2) of venous blood in vivo. Using an interecho spacing of 25 ms, the results on visual activation studies in eight volunteers showed T(2)(CPMG) values increasing from 128 +/- 9 ms to 174 +/- 18 ms upon activation, corresponding to local OER values of 0.38 +/- 0.04 and 0.18 +/- 0.05 during baseline activity and visual stimulation, respectively. These OER values are in good agreement with literature data on venous oxygenation and numbers determined previously using a single-echo approach, while the measured T(2)s are about 20-40 ms longer. PMID- 11477632 TI - Bicuculline-induced brain activation in mice detected by functional magnetic resonance imaging. AB - Dynamic measurements of local changes in relative cerebral blood volume (CBV(rel)) during a pharmacological stimulation paradigm were performed in mice. Using magnetite nanoparticles as an intravascular contrast agent, high-resolution CBV(rel) maps were obtained. Intravenous administration of the GABA(A) antagonist bicuculline prompted increases in local CBV(rel) as assessed by MRI with a high spatial resolution of 0.2 x 0.2 mm(2) and a temporal resolution of 21 s. Signal changes occurred 20-30 s after the onset of drug infusion in the somatosensory and motor cortex, followed by other cortical and subcortical structures. The magnitudes of the CBV(rel) increases were 18% +/- 4%, 46% +/- 14%, and 67% +/- 7%, as compared to prestimulation values for the cortex, and 9% +/- 3%, 25% +/- 4%, and 36% +/- 7% for the caudate putamen for bicuculline doses of 0.6, 1.25, and 1.5 mg/kg, respectively. On-line monitoring of transcutaneous carbon dioxide tension PtcCO(2) reflecting arterial PaCO(2) did not show any alteration during the stimulation paradigm. One of five of the mice receiving the highest bicuculline dose, and three of seven receiving the intermediate dose displayed a different cortical response pattern. After a CBV(rel) increase of 40% lasting for approximately 1 min, significant CBV(rel)reductions by 80% have been observed. Subcortical structures did not display this behavior. The present study suggests that this noninvasive approach of functional MRI (fMRI) can be applied to study drug-induced brain activation by central nervous system (CNS) drugs in mice under normal and pathological situations. PMID- 11477633 TI - Effect of black blood MR image quality on vessel wall segmentation. AB - Black blood MRI has become a popular technique for measuring arterial wall area as an indicator of plaque size. Computer-assisted techniques for segmenting vessel boundaries have been developed to increase measurement precision. In this study, the carotid arteries of four normal subjects were imaged at seven different fields of view (FOVs), keeping all other imaging parameters fixed, to determine whether spatial resolution could be increased at the expense of image quality without sacrificing precision. Wall areas were measured via computer assisted segmentation of the vessel boundaries performed repeatedly by two operators. Analysis of variance (ANOVA) demonstrated that the variability of wall area measurements was below 1.5 mm(2) for in-plane spatial resolutions between 0.22 mm and 0.37 mm. An inverse relationship between operator variability and the signal difference-to-noise ratio (SDNR) demonstrated that semi-automatic segmentation of the wall boundaries was robust for SDNR >3, defining a criterion above which subjective image quality can be degraded without an appreciable loss of information content. Our study also suggested that spatial resolutions higher than 0.3 mm may be required to quantify normal wall areas to within 10% accuracy, but that the reduced SNR associated with the higher resolution may be tolerated by semi-automated wall segmentation without an appreciable loss of precision. PMID- 11477634 TI - Determination of skeletal muscle perfusion using arterial spin labeling NMRI: validation by comparison with venous occlusion plethysmography. AB - T(1)-based determination of perfusion was performed with the high temporal and spatial resolution that monitoring of exercise physiology requires. As no data were available on the validation of this approach in human muscles, T(1)-based NMRI of perfusion was compared to standard strain-gauge venous occlusion plethysmography performed simultaneously within a 4 T magnet. Two different situations were investigated in 21 healthy young volunteers: 1) a 5-min ischemia of the leg, or 2) a 2-3 min ischemic exercise consisting of a plantar flexion on an amagnetic ergometer. Leg perfusion was monitored over 5-15 min of the recovery phase, after the air-cuff arterial occlusion had been released. The interesting features of the sequence were the use of a saturation-recovery module for the introduction of a T(1) modulation and of single-shot spin echo for imaging. Spatial resolution was 1.7 x 2.0 mm and temporal resolution was 2 s. For data analysis, ROIs were traced on different muscles and perfusion was calculated from the differences in muscle signal intensity in successive images. To allow comparison with the global measurement of perfusion by plethysmography, the T(1) based NMR measurements in exercising muscles were rescaled to the leg cross section. The perfusion measurements obtained by plethysmography and NMRI were in close agreement with a correlation coefficient between 0.87 and 0.92. This indicates that pulsed arterial techniques provide determination of muscle perfusion not only with superior spatial and temporal resolution but also with exactitude. PMID- 11477635 TI - Optimal dose of Gd-DTPA in dynamic MR studies. AB - The relationship between the administered dose d of Gd-DTPA and the accuracy of measurements of the glomerular filtration rate G and the cardiac output O was investigated. For a wide range of values the concentration of Gd-DTPA can be uniquely derived from MR signals and precontrast longitudinal relaxation time. Fixed and random errors in these measured variables were analyzed. Depending on noise level and the level of renal function, errors in G reach a minimum for d = 1.4-2.8 mmol. Random errors in G are relatively insensitive to d as long as d > 1.5 mmol. These results establish the feasibility of dynamic MR measurements using doses of Gd-DTPA that are several times lower than current standards. PMID- 11477636 TI - Techniques for fast stereoscopic MRI. AB - Stereoscopic MRI can impart 3D perception with only two image acquisitions. This economy over standard multiplanar 3D volume renderings allows faster frame rates, which are needed for real-time imaging applications. Real-time 3D perception may enhance the appreciation of complex anatomical structures, and may improve hand eye coordination while manipulating a medical device during an image-guided interventional procedure. To this goal, a system is being developed to acquire and display stereoscopic MR images in real-time. A clinically used, fast gradient recalled echo-train sequence has been modified to produce stereo image pairs. Features have been added for depth cueing, view sharing, and bulk signal suppression. A workstation was attached to a clinical MR scanner for fast data extraction, image reconstruction and stereoscopic image display. PMID- 11477637 TI - Imaging longitudinal cardiac strain on short-axis images using strain-encoded MRI. AB - This article presents a new method for measuring longitudinal strain in a short axis section of the heart using harmonic phase magnetic resonance imaging (HARP MRI). The heart is tagged using 1-1 SPAMM at end-diastole with tag surfaces parallel to a short-axis imaging plane. Two or more images are acquired such that the images have different phase encodings in a direction orthogonal to the image plane. A dense map of the longitudinal strain can be computed from these images using a simple, fast computation. Simulations are conducted to study the effect of noise and the choice of out-of-plane phase encoding values. Longitudinal strains acquired from a normal human male are shown. PMID- 11477638 TI - Ghost artifact cancellation using phased array processing. AB - In this article, a method for phased array combining is formulated which may be used to cancel ghosts caused by a variety of distortion mechanisms, including space variant distortions such as local flow or off-resonance. This method is based on a constrained optimization, which optimizes SNR subject to the constraint of nulling ghost artifacts at known locations. The resultant technique is similar to the method known as sensitivity encoding (SENSE) used for accelerated imaging; however, in this formulation it is applied to full field-of view (FOV) images. The method is applied to multishot EPI with noninterleaved phase encode acquisition. A number of benefits, as compared to the conventional interleaved approach, are reduced distortion due to off-resonance, in-plane flow, and EPI delay misalignment, as well as eliminating the need for echo-shifting. Experimental results demonstrate the cancellation for both phantom as well as cardiac imaging examples. PMID- 11477639 TI - IMPACT: image-based physiological artifacts estimation and correction technique for functional MRI. AB - Functional MRI (fMRI) signal variation induced by respiratory and cardiac motion affects the activation signal and limits the accuracy of analysis. Current physiological motion correction methods require either synchronization with external monitoring of respiration and heartbeat, specialized pulse sequence design, or k-space data. The IMage-based Physiological Artifacts estimation and Correction Technique (IMPACT), which is free from these constraints, is described. When images are acquired fast enough to sample physiological motion without aliasing, respiratory and cardiac signals can be directly estimated from magnitude images. Physiological artifacts are removed by reordering images according to the estimated respiratory and cardiac phases and then subtracting the Fourier-fitted variation from magnitude images. Compared with the k-space based method, this method can efficiently and effectively reduce the overall signal fluctuation in the brain and increase the activated area. With this new technique, physiological artifacts can be reduced using traditional fMRI pulse sequences, and existing data can be corrected and reanalyzed without additional experiments. PMID- 11477640 TI - FLAIR imaging using nonselective inversion pulses combined with slice excitation order cycling and k-space reordering to reduce flow artifacts. AB - High-signal artifacts produced by cerebrospinal fluid (CSF) flow can adversely affect fluid-attenuated inversion recovery (FLAIR) imaging of the brain and spinal cord. This study explores the use of a nonslice-selective inversion pulse to eliminate CSF flow artifacts together with a technique called "K-space Reordered by Inversion-time for each Slice Position" (KRISP) to achieve constant contrast in a multislice acquisition. Theory shows that with this method the CSF point spread function (PSF) has a minimum at the center and attenuated side lobes, providing CSF suppression, but residual edge signals remain. The PSF for brain is only mildly attenuated and signals for extended regions are not attenuated. KRISP FLAIR sequences were assessed in 15 patients (10 brain and five spinal cord cases). The images showed reduced CSF and blood flow artifacts and higher conspicuity of the cortex, meninges, ventricular system, brainstem, and cerebellum when compared with conventional FLAIR sequences. PMID- 11477641 TI - Contrast-enhanced magnetic resonance angiography of the iliac arteries: optimization by injection simulation. AB - Protocols for contrast-enhanced magnetic resonance angiography (CE-MRA) of the iliac arteries were optimized by computer simulations based on an impulse response function (IRF) of contrast agent (CA) concentration as a function of time obtained for 20 patients. Protocols with sequential, centric, and elliptical k-space coverage, different repetition rates (5 and 10 ms), and CA doses (0.1, 0.2, and 0.3 mmol/kg b.w.) were compared in terms of signal-to-noise ratio (SNR), distortion of vessel profiles, and sensitivity to timing errors. IRF-based simulations successfully characterized CA recirculation. Slow-rate CA infusions were found to achieve relatively high enhancement. In terms of SNR, there is no advantage in increasing the repetition rate. Distortion of vessel profiles is more likely in elliptic and centric k-space coverage. Protocols based on sequential k-space coverage and relatively long CA infusions proved to be particularly suited to large-FOV iliac examinations as they are relatively insensitive to timing errors. PMID- 11477642 TI - Bayesian source separation for reference function determination in fMRI. AB - In analyzing fMRI results, identification of significant activation in voxels is a crucial task. A standard method selects a "known" reference function and performs a regression of the time courses on it and a linear trend. Once the linear trend is found, the correlation between the assumed to be known reference function and the detrended observed time-course in each voxel is computed. But the most important question is: How does one choose the reference function? Here, a Bayesian source separation approach to determining the underlying reference function is described and applied to real fMRI data. This underlying reference function is the unobserved response due to the presentation of the experimental stimulus. PMID- 11477643 TI - Radio frequency magnetic field mapping of a 3 Tesla birdcage coil: experimental and theoretical dependence on sample properties. AB - The RF B(1) distribution was studied, theoretically and experimentally, in phantoms and in the head of volunteers using a 3 T MRI system equipped with a birdcage coil. Agreement between numerical simulation and experiment demonstrates that B(1) distortion at high field can be explained with 3D full-Maxwell calculations. It was found that the B(1) distribution in the transverse plane is strongly dependent on the dielectric properties of the sample. We show that this is a consequence of RF penetration effects combined with RF standing wave effects. In contrast, along the birdcage coil z-axis the B(1) distribution is determined mainly by the coil geometry. In the transverse plane, the region of B(1) uniformity (within 10% of the maximum) was 15 cm with oil, 6 cm with distilled water, 11 cm with saline, and 10 cm in the head. Along z the B(1) uniformity was 9 cm with phantoms and 7 cm in the head. PMID- 11477644 TI - A comparison between human magnetostimulation thresholds in whole-body and head/neck gradient coils. AB - Gradient coil magnetostimulation thresholds were measured in a group of 20 volunteers in both a whole-body gradient coil and a head/neck gradient coil. Both coils were operated using both x and y axes simultaneously (xy oblique mode). The waveform applied was a 64-lobe trapezoidal train with 1-ms flat-tops and varying rise times. Thresholds were based on the subjects' perception of stimulation, and painful sensations were not elicited. Thresholds were expressed in terms of the total gradient excursion required to cause stimulation as a function of the duration of the excursion. Thresholds for each subject were fit to a linear model, and values for the threshold curve slope (SR(min)) and vertical axis intercept (DeltaG(min)) were extracted. For the body coil, the mean values were: SR(min) = 62.2 mT/m/ms, DeltaG(min) = 44.4 mT/m. For the head/neck coil, the mean values were: SR(min) = 87.3 mT/m/ms, DeltaG(min) = 78.9 mT/m. These curve parameters were combined with calculated values for the induced electric field as a function of position within the coil to yield the tissue specific parameters E(r) (electric field rheobase) and tau(c) (chronaxie). For tissue stimulated within the body coil, the mean values were: E(r) = 1.8 V/m, tau(c) = 770 micros. For tissue stimulated within the head/neck coil, the mean values were: E(r) = 1.3 V/m, tau(c) = 1100 micros. Scalar potential contributions were not included in the calculation of induced electric fields. The mean threshold curves were combined with the gradient system performance curves to produce operational limit curves. The operational limit curves for the head/neck coil system were verified to be higher than those of the whole-body coil; however, the head/neck system was also found to be physiologically limited over a greater range of its operation than was the body coil. Subject thresholds between the two coils were not well correlated. PMID- 11477645 TI - Restoration of motion-related signal loss and line-shape deterioration of proton MR spectra using the residual water as intrinsic reference. AB - A postprocessing method to restore motion-related signal loss and line-shape deterioration in single-volume proton MR spectroscopy (MRS) is presented. Each acquisition is corrected by its phase offset and frequency shift, extracted from the residual water signal prior to averaging. Requirements are good gradient selection and selective suppression of CSF in residual water. Stimulated echo acquisition mode (STEAM) spectra (TE = 30 ms) were analyzed using the LCModel program to study gains in metabolite signal and spectral quality in five brain regions. Increases of total N-acetyl-aspartate concentrations of up to 5% were observed. The method may be beneficial for clinical examinations of less compliant subjects and for dynamic spectroscopy. PMID- 11477646 TI - Inverse T(2) contrast at 1.5 Tesla between gray matter and white matter in the occipital lobe of normal adult human brain. AB - T(2) of cortical gray matter is generally assumed to be longer than that of white matter. It is shown here that this is not the case in the occipital lobe, but that this effect is often obscured at lower resolution and concealed in standard T(2)-weighted images. Using a high-resolution (1 x 1.3 x 2 mm(3)) segmented EPI Carr-Purcell-Meiboom-Gill sequence, T(2) relaxation times of the brain were measured at 1.5 T for eight healthy adult volunteers. The average T(2) values of cortical gray and white matter were found to be 88 +/- 2 and 84 +/- 3 msec in the frontal lobe, 84 +/- 2 and 83 +/- 3 msec in the parietal lobe, and 79 +/- 1 and 87 +/- 3 msec in the occipital lobe, respectively. This unexpected occipital T(2) contrast between gray and white matter is attributed to regional differences in iron concentration. PMID- 11477647 TI - Single-shot EPI with signal recovery from the susceptibility-induced losses. AB - A major problem in the gradient-recalled echo-planar imaging (EPI) method that also uses a long echo time (TE) is the severe signal loss in regions with large static field inhomogeneities. These regions include the ventral frontal, medial temporal, and inferior temporal lobes, which experience inhomogeneities induced by susceptibility effects commonly found near air/tissue interfaces. For functional magnetic resonance imaging (fMRI) studies that use both gradient recalled EPI at relatively long TE and high-field scanners, this signal loss is severe, preventing investigation of certain human cognitive processes that involve these regions, such as memory and attention. Methods have been developed to recover this signal loss; however, most of them require multiple excitations and thus compromise temporal resolution. In this report, a new technique is described which achieves good signal recovery within a single excitation. It is anticipated that this technique will prove useful for fMRI studies in inhomogeneous areas that require high temporal resolution. PMID- 11477648 TI - Measuring the human retinal oxygenation response to a hyperoxic challenge using MRI: eliminating blinking artifacts and demonstrating proof of concept. AB - The retinal oxygenation response to a hyperoxic challenge measured using MRI appears to be an early and accurate marker of retinopathy risk in experimental models, with promising clinical potential. However, the application of this technique in humans is limited by blinking artifacts that can confound detection of subtle signal intensity changes. We asked subjects to refrain from blinking during a 12-s fast low-angle shot (FLASH) image, and to blink if needed during the following 3-s rest period. This no-blink blink cycle was repeated sequentially 20 times during either room-air or 100% oxygen breathing. Significant change (P < 0.05) was detected for the first time from the resultant blinking-artifact-free images in the preretinal vitreous oxygen tension (upper limit of about 13 mm Hg (1.8 KPa, N = 3)) following a 10-min hyperoxic inhalation challenge. These results provide the proof-of-concept data needed for future MRI evaluation of the retinal oxygenation response and human retinopathy, such as diabetic retinopathy. PMID- 11477649 TI - MR angiographic investigation of transient focal cerebral ischemia in rat. AB - Contrast agent free time-of-flight magnetic resonance angiography (TOF-MRA) was applied to the intraluminal thread occlusion model of experimental stroke in rat. It was combined with perfusion- and diffusion-weighted imaging (PWI and DWI) sequences to correlate occlusion and reopening of the middle cerebral artery with alterations in these well-established magnetic resonance sequences. Since TOF-MRA can be repeated without limitations, the time course of vascular patency is demonstrated during an experimental period of up to 8 h (2 h control, 1 h ischemia, 3-6 h reperfusion). With an acquisition time of 10 min, TOF-MRA proved to be suitable to analyze the vascular state of occlusion and reperfusion repetitively in longitudinal studies. Spatial resolution was sufficient to observe neurovascular structural details. In eight out of 10 animals complete vessel occlusion by the intraluminal thread could be validated by an entirely extinguished signal of the ipsilateral middle cerebral artery (MCA) in the angiograms. This was in accordance with a perfusion deficit in the MCA vascular territory detected by PWI (reduction to 30.4 +/- 7.4% relative to contralateral side) and a disturbance of water ion homeostasis monitored by DWI in this area. One animal showed a delayed occlusion after 30 min of MCA occlusion, in another animal vessel occlusion failed. In seven out of the eight successful occlusion experiments there was immediate reperfusion after withdrawal of the thread. One animal showed a delayed reperfusion after suture retraction. Remarkable hemispheric differences in vascular branching of the MCA could be recognized in three out of 10 animals. In conclusion, TOF-MRA is considered a helpful method to survey even in small laboratory animals the correct time course of vascular occlusion and reopening in experimental ischemia, and provides complementary information to the tissue perfusion status monitored by PWI and the ischemic lesion territory detected by DWI. PMID- 11477650 TI - MRI of lung parenchyma in rats and mice using a gradient-echo sequence. AB - Signal of lung parenchymal tissue from the living rat and mouse lung was detected at 4.7 T with a good signal-to-noise ratio and motion-suppressed artifacts using a short TE gradient-echo sequence. Neither cardiac nor respiratory gating were applied, and animals respired freely during data collection. Mean T(2)* relaxation times of parenchyma in the anterior, middle and posterior regions of both lungs ranged between 403 and 657 micros and 397 and 751 micros, respectively for the rat and mouse. For the rat in the prone position, there was a gradient in T(2)* values, from the posterior to the anterior regions of both lungs. In the supine position, however, T(2)* values were larger in the posterior and in the anterior portions. For the mouse in both prone and supine positions, there was a tendential gradient in T(2)* from the anterior to the posterior portions. The robustness of the approach renders it well suited for routine applications, e.g. in pharmacological studies concerning asthma models in small rodents. The method was applied to lung inflammation models involving challenge with ovalbumin or lipopolysaccharide. PMID- 11477651 TI - 1H- and (31)P-MR spectroscopy of primary and recurrent human brain tumors in vitro: malignancy-characteristic profiles of water soluble and lipophilic spectral components. AB - In vitro NMR spectroscopy was performed on specimen of human brain tumors. From all patients, tissue samples of primary tumors and their first recurrences were examined. (31)P- and (1)H-spectra were recorded from samples of meningioma, astrocytoma and glioblastoma. A double extraction procedure of the tissue samples permitted acquisition of information from the membrane fraction and from the cytosolic fraction. (31)P-spectra were used to analyze the lipophilic fraction (phospholipids of the membrane) of the tissue extracts, while the (1)H-spectra reflected information on the metabolic alterations of the hydrophilic, cytosolic fraction of the tissue. The tumor types showed distinctive spectral patterns in both the (31)P- and the (1)H-spectra. Based on the total detectable (31)P signal, the level of phosphatidylcholine was about 34% lower in primary astrocytomas than in primary glioblastomas (p = 0.0003), whereas the level of sphingomyelin was about 45% lower in primary glioblastomas than in primary astrocytomas (p = 0.0061). A similar tendency of these phospholipids was observed when comparing primary and recurrent astrocytoma samples from the same individuals [+15% (p = 0.0103) and -23% (p = 0.0314) change, respectively]. (1)H-spectra of gliomas were characterized by an increase of the ratios of alanine, glycine and choline over creatine as a function of the degree of malignancy. In agreement with findings in the (31)P-spectra, the (1)H-spectra of recurrent astrocytomas showed metabolic profiles of increased malignancy in comparison to their primary occurrence. Since gliomas tend to increase in malignancy upon recurrence, this may reflect evolving tumor metabolism. (1)H-spectra of meningiomas showed the highest ratio of alanine over creatine accompanied by a near absence of myo-inositol. Phospholipid profiles of meningiomas showed higher fractional contents of phosphatidylcholine along with lower phosphatidylserine compared to astrocytomas, while higher phosphatidylethanolamine and sphingomyelin fractional contents distinguished meningiomas from glioblastomas. The extraction method being used in this study combined with high-resolution (1)H- and (31)P-MRS provides a wide range of biochemical information, which enables differentiation not only between tumor types but also between primary and recurrent gliomas, reflecting an evolving tumor metabolism. PMID- 11477652 TI - Peripheral somatosensory fMRI in mouse at 11.7 T. AB - The feasibility of performing extremely-high resolution somatosensory fMRI in anesthetized mice using BOLD contrast at 11.7 T was investigated. A somatosensory stimulus was applied to the hindlimb of an alpha-chlorolose anesthetized mouse resulting in robust (p < 4 x 10(-3)) BOLD changes in somatosensory cortex and large veins. Percentage modulation of the MR signal in cortex exceeded 7%. Experiments that artificially modulated the inspired oxygen tension were also conducted; the results revealed large, heterogeneous, BOLD contrast changes in the mouse brain. In addition, T(1), T(2), and T(2)* values in gray matter at 11.7 T were evaluated. Discussion of the sensitivity limitations of BOLD fMRI in the tiny mouse central nervous system is presented. These methods show promise for the assessment of neurological function in mouse models of CNS injury and disease. PMID- 11477653 TI - Proton T (1) and T (2) relaxation times of human brain metabolites at 3 Tesla. AB - Longitudinal and transverse relaxation times were measured for proton MRS signals from human brain metabolites at 3 T using a short-echo STEAM protocol and a surface coil as a transmitter/receiver. Volumes of interest containing mostly grey or mostly white matter were selected in occipital lobes of healthy subjects and relaxation times for the following resonances were obtained: N acetylaspartate at 2.01 ppm (T(1) and T(2)), glutamate at 2.35 ppm (T(1)), creatine at 3.03 and 3.92 ppm (T(1) and T(2)), choline-containing substances at 3.22 ppm (T(1) and T(2)), myo-inositol at 3.57 and 3.65 ppm (T(1)) and the overlapping signals of glutamate and glutamine at 3.75 ppm (T(1)). The T(1) relaxation times obtained range from 0.97 to 1.47 s for grey matter and from 0.87 to 1.35 s for white matter. On the other hand, T(2) relaxation times range from 116 to 247 ms and from 141 to 295 ms in grey and white matter, respectively. Generally, the T(1) values measured at 3 T are close to the previously published data found at 1.5, 2 and 4.1 T. Also, the T(2) values confirm the previously observed decrease in transverse relaxation times with increasing static magnetic field. The proton relaxation times obtained will allow improved sequence design and spectra quantitation at 3 T, currently tested for enhanced clinical applications. PMID- 11477655 TI - t(10;11)-acute leukemias with MLL-AF10 and MLL-ABI1 chimeric transcripts: specific expression patterns of ABI1 gene in leukemia and solid tumor cell lines. AB - The recurrent translocation t(10;11) is associated with acute myeloid leukemia (AML). The AF10 gene on chromosome 10 at band p12 and MLL at 11q23 fuse in the t(10;11)(p12;q23). Recently, we have identified ABI1 as a new partner gene for MLL in an AML patient with a t(10;11)(p11.2;q23). The ABI1 is a human homologue of the mouse Abl-interactor 1 (Abi1), encoding an Abl-binding protein. The ABI1 protein exhibits sequence similarity to homeotic genes, and contains several polyproline stretches and a src homology 3 (SH3) domain. To clarify the clinical features of t(10;11)-leukemias, we investigated 6 samples from acute leukemia patients with t(10;11) and MLL rearrangement and detected MLL-AF10 chimeric transcripts in 5 samples and MLL-ABI1 in one. The patient with MLL-ABI1 chimeric transcript is the second case described, thus confirming that the fusion of the MLL and ABI1 genes is a recurring abnormality. Both of the patients with MLL-ABI1 chimeric transcript are surviving, suggesting that these patients have a better prognosis than the patients with MLL-AF10. To investigate the roles of AF10 and ABI1 further, we examined the expression of these genes in various cell lines and fresh tumor samples using the reverse transcriptase-polymerase chain reaction method. Although AF10 was expressed in almost all cell lines similarly, the expression patterns of ABI1 were different between leukemia and solid tumor cell lines, suggesting the distinctive role of each isoform of ABI1 in these cell lines. We also determined the complete mouse Abi1 sequence and found that the sequence matched with human ABI1 better than the originally reported Abi1 sequence. Further functional analysis of the MLL-AF10 and MLL-ABI1 fusion proteins will provide new insights into the leukemogenesis of t(10;11)-AML. PMID- 11477656 TI - Amplification of ribosomal RNA genes in acute myeloid leukemia. AB - Gene amplification is a relatively rare event in acute myeloid leukemia (AML). Double minutes (dmin) and homogeneously staining regions are well established phenomena as cytogenetic correlates of gene amplification. Recently, however, two additional mechanisms leading to gene amplification, i.e., segmental jumping translocations and formation of ring chromosomes, have been described. We report four patients with AML, in whom bone marrow cells exhibited amplifications of ribosomal RNA (rRNA) genes in the form of ring chromosomes or a hsr. In two patients, the MLL gene, and in one patient the CBFA2 gene were shown to be co amplified with rRNA genes. In two of the four patients, multiple copies of alpha satellite sequences of the centromeres 13 and 21, respectively, were also demonstrated. In three of the four patients, the clinical course was very aggressive, leading to death within 2-8 months. In these three patients, complex karyotype abnormalities were found, whereas the karyotype of Patient 4 was characterized only by supernumerary ring 21 chromosomes of different sizes and a trisomy 8 in half of the metaphases. Modes of origin and clinical significance of the amplification of rRNA genes are discussed. PMID- 11477657 TI - Detection of three novel translocations and specific common chromosomal break sites in malignant melanoma by spectral karyotyping. AB - Chromosomal aberrations in malignant melanoma cells have been reported using standard chromosome banding analysis and comparative genomic hybridization. To identify marker chromosomes and translocations that are difficult to characterize by standard banding analysis, 15 early passage malignant melanoma cell lines were examined using spectral karyotyping. All 15 tumor cell lines had lost all or part of 1p and 10q. Losses of material on chromosome arms 4p (12/15), 6q (12/15), 9p (15/15), 12p (13/15), 12q (13/15), 13q (11/15), and 19q (14/15) were the next most frequent events. Gain of chromosome arms 1q (11/15), 6p (13/15), and 20q11 (14/15) was also observed. Interestingly, we identified translocations der(12)t(12;20)(q15;q11), der(19)t(10;19)(q23;q13), and der(12)t(12;19)(q13;q13) in 4/15 tumors. Three recurring translocations involving four of the most frequent break points were detected. The identification of recurring translocations and unique chromosome break points in melanoma will aid in the identification of the genes that are important in the neoplastic process. PMID- 11477658 TI - Molecular cytogenetic definition of three distinct chromosome arm 14q deletion intervals in gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) are mesenchymal neoplasms characterized by frequent chromosome arm 14q losses. In this study, the 14q changes in a series of 39 histologically and immunohistochemically confirmed GISTs were analyzed in detail by metaphase and/or interphase fluorescence in situ hybridization (FISH) studies using 21 genetically well-characterized, region-specific 14q11-24 YAC clones. By conventional cytogenetic analysis, acquired clonal chromosome aberrations were found in 17 out of 35 tumors. Chromosome 14 was involved in 13 cases; six specimens showed complete chromosome 14 loss, while the remaining seven had structural abnormalities with the breakpoints residing within the intervals 14q11-13 or 14q22-24. Other recurrent chromosome aberrations included frequent deletions of chromosome 1p (11/17), losses of chromosome 22 (7/17), losses or deletions of chromosome arm 13 (6/17) or 15 (4/17), and gains or translocations involving chromosome 17 (4/17). Combining cytogenetic data with double-color FISH analysis, total or partial losses of 14q material were detected in 29 out of 36 tumors (81%). The 14q losses were found in all stages and histological subtypes. Two most frequent common deletion regions flanked by YACs 931B1 and 761D4, and 802E7 and 892C11 at 14q23-24 (25/30 of each; 83%) could be identified. Furthermore, 21 tumors (70%) shared a region of deletion defined by YACs 957H10 and 931E5 at 14q11-12. Our results suggest the presence of at least three distinct critical deletion regions on chromosome 14 in GISTs. PMID- 11477659 TI - Identification of tumor metastasis suppressor region on the short arm of human chromosome 20. AB - Acquisition of metastatic ability by prostate cancer cells is the hallmark of their lethal trait and outcome. However, the genetic alterations underlying the clinical progression and pathogenesis of prostate cancer are not well understood. Several studies involving loss of heterozygosity (LOH) and comparative genomic hybridization analysis have identified distinctively altered regions on various human chromosomes, and genomic imbalance of chromosome 20 was implicated in progression and recurrence of prostate tumors. To examine the role of chromosome 20 in prostate neoplasms, we introduced this chromosome into highly metastatic rat prostate cancer cells using the microcell-mediated chromosome transfer technique. Introduction of the chromosome resulted in significant suppression of the metastatic ability of the hybrid cells, by as much as 98%, without any interference with the in vivo growth rate or tumorigenicity of primary tumor in SCID mice. Our STS-PCR analysis on 10 hybrid clones indicates that the suppressor activity of chromosome 20 is located in the p11.23-12 region. Further examination of the hybrid clones by experimental metastasis assay and histologic analysis as well as Matrigel invasion assay suggests the involvement of the suppressor region at an early stage of invasion and extravasation. We also investigated the status of the chromosome 20 suppressor region in pathology specimens from human prostate cancer patients and detected the frequent loss of this region in high-grade tumors. These results suggest the presence of a putative suppressor gene on human chromosome 20 that is functionally involved in development of prostate cancer metastases. PMID- 11477660 TI - Gain of 1q and loss of 9q21.3-q32 are associated with a less favorable prognosis in papillary thyroid carcinoma. AB - In order to approach the genetic mechanisms behind initiation and progression of papillary thyroid carcinoma (PTC) tumorigenesis, we characterized numerical chromosomal imbalances in a panel of 25 PTCs with varying histopathological and clinical features using comparative genomic hybridization (CGH). The most frequently detected imbalance was gain of 9q33-qter, which was seen in close to 30% of the cases. The commonly occurring regions of loss were assigned to 22q (12%) and 9q21.3-q32 (12%), while gains preferentially involved the entire X chromosome (20%), 1q (16%), 17q (16%), and 22q (12%). The distribution of CGH alterations supports the idea of a progression of genetic events in the development of PTC, where gain of 9q33-qter would represent a relatively early event that is followed by loss of 22q and gain of X, 1q, 17q, and 22q. When the detected CGH alterations were compared with the clinical outcome and the histopathological features of the 25 PTC cases, several statistically significant correlations were revealed. The total number of genetic alterations was higher in tumors from patients with aggressive disease as compared to those without signs of aggressiveness. Gain of 1q and loss of 9q21.3-q32 were exclusively seen in tumors from patients with aggressive disease, and the presence of distant metastases was associated with gain of 1q. A sex-dependent distribution was also evident for one of the common alterations, with gain of X exclusively seen in male cases. Taken together, the findings identify several candidate locations for tumor suppressor genes and oncogenes that are potentially involved in the establishment and progression of papillary thyroid carcinogenesis. PMID- 11477661 TI - Chromosomal imbalances in gastric and esophageal adenocarcinoma: specific comparative genomic hybridization-detected abnormalities segregate with junctional adenocarcinomas. AB - The incidence of adenocarcinoma arising at the esophagogastric junction (EGJ) is increasing at a rate greater than that for any other form of solid malignancy. Commensurate with this, the incidence of histologically similar tumors arising in the gastric body and antral mucosa is declining. The increased incidence of the proximal group of tumors may reflect, in part, the higher prevalence of Barrett esophagus. These epidemiological features suggest that histologically similar tumors arising at the EGJ and from the distal stomach are different, which may be reflected in the genetic abnormalities that characterize the two groups of tumors. The purpose of this study was to screen genomic DNA from adenocarcinomas of the esophagus and stomach for regions of chromosomal imbalance, using comparative genomic hybridization to determine whether tumors at the EGJ (junctional tumors) have a different profile compared with tumors of the distal stomach. Tumor samples were derived from a series of 48 gastroesophageal adenocarcinomas (20 junctional and 28 distal) that were acquired prospectively from patients undergoing esophagogastrectomy. These tumors are characterized by several regions of chromosomal imbalance with no obvious correlation between most regions of abnormal copy number and tumor type. However, our study shows for the first time cytogenetic abnormalities (5p+ and 18q-) that identify statistically significant differences (P < 0.02 and < 0.05, respectively) between junctional and distal gastric tumors. These differences are gain of 5p (55% [11/20] of junctional tumors vs. 21% [6/28] of distal gastric tumors) and loss of 18q (25% [5/20] cases of junctional tumors vs. 4% [1/28] of distal tumors) segregating with tumors of the EGJ. These abnormalities may distinguish distinct tumor subtypes that are recognized in epidemiological and clinical studies but that are otherwise histologically identical. PMID- 11477662 TI - Gain of 1q and loss of 22 are the most common changes detected by comparative genomic hybridisation in paediatric ependymoma. AB - Ependymomas are the third most common brain tumour in the paediatric population. Although cytogenetic and molecular analyses have pinpointed deletions of chromosomes 6q, 17, and 22 in a subset of tumours, definitive patterns of genetic aberrations have not been determined. In the present study, we analysed 40 ependymomas from paediatric patients for genomic loss or gain using comparative genomic hybridisation (CGH). Eighteen of the tumours (45%) had no detectable regions of imbalance. In the remaining cases, the most common copy number aberrations were loss of 22 (25% of tumours) and gain of 1q (20%). Three regions of high copy number amplification were noted at 1q24-31 (three cases), 8q21-23 (two cases), and 9p (one case). Although there was no association with the loss or gain of any chromosome arm or with benign versus anaplastic histologic characteristics, the incidence of gain of 7q and 9p and loss of 17 and 22 was significantly higher in recurrent versus primary tumours. This study has identified a number of chromosomal regions that may contain candidate genes involved in the development of different subgroups of ependymoma. PMID- 11477663 TI - High-density allelotype of the commonly studied gastric cancer cell lines. AB - Gastric cancer is one of the leading causes of death from cancer throughout the world, and studies to elucidate the genetic defects found in this type of cancer are growing in number. Increasingly sophisticated techniques and the sequencing of the human genome have had an impact on the scope of such studies. While the use of tumor specimens remains popular, more emphasis is being placed on cell lines as model systems where specific data can be directly combined with results from other studies. This article describes a genetic survey of the most widely used gastric adenocarcinoma cell lines. The allelotype at 351 polymorphic loci in 14 cell lines was obtained, and the results from the 4,900 polymerase chain reactions are displayed. In addition to confirming loss of heterozygosity on chromosome arms 6p, 7q, 17p, and 18, additional deletions on arm 5p and the pericentromeric regions of chromosomes 1 and 10 were detected. Areas that might contain homozygous deletions or amplifications also were mapped. The rate of microsatellite instability was quantified and shown to vary greatly among the different cell lines. Most important, this study serves as a genetic scaffold for the integration of past and future studies on the nature of the genetic defects in gastric cancer. PMID- 11477664 TI - An ins(X;11)(q24;q23) fuses the MLL and the Septin 6/KIAA0128 gene in an infant with AML-M2. AB - The MLL (HRX, ALL-1 HTRX) gene at chromosome band 11q23 frequently is rearranged in acute lymphoblastic and myeloblastic leukemia. To date, more than 40 different 11q23 abnormalities have been described on the cytogenetic level, and at least 25 of the respective fusion partner genes are cloned. The vast majority of the respective reciprocal translocations generate a chimeric 5'-MLL/partner-3' gene on the derivative 11q23. In this work, we report a unique ins(X;11)(q24;q23) in an infant with acute myeloid leukemia (AML-M2) that fuses the human KIAA0128 gene at Xq24 with MLL. In contrast to the typical reciprocal MLL translocations, however, we provide evidence that the 5'-MLL/KIAA0128-3' fusion resides on Xq24 rather than on 11q23. The KIAA0128 gene encodes the human Septin 6 protein, which contains an ATP-GTP binding motif and three nuclear targeting sequences in its carboxy terminus. The maintenance of the reading frame of the 5'-MLL/KIAA0128-3' mRNA fusion allows for the formation of a novel chimeric protein. Septin 6 is the third member of the Septins that is fused to the MLL protein; the other two are hCDCrel at 22q11 and MSF at 17q25. PMID- 11477665 TI - Mutation analysis of the CDKN2A promoter in Australian melanoma families. AB - Approximately 50% of all melanoma families worldwide show linkage to 9p21-22, but only about half of these have been shown to contain germ line CDKN2A mutations. It has been hypothesized that a proportion of these families carry mutations in the noncoding regions of CDKN2A. Several Canadian families have been reported to carry a mutation in the 5' UTR, at position -34 relative to the start site, which gives rise to a novel AUG translation initiation codon that markedly decreases translation from the wild-type AUG (Liu et al., 1999). Haplotype sharing in these Canadian families suggested that this mutation is of British origin. We sequenced 1,327 base pairs (bp) of CDKN2A, making up 1,116 bp of the 5' UTR and promoter, all of exon 1, and 61 bp of intron 1, in at least one melanoma case from 110 Australian families with three or more affected members known not to carry mutations within the p16 coding region. In addition, 431 bp upstream of the start codon was sequenced in an additional 253 affected probands from two-case melanoma families for which the CDKN2A mutation status was unknown. Several known polymorphisms at positions -33, -191, -493, and -735 were detected, in addition to four novel variants at positions 120, -252, -347, and -981 relative to the start codon. One of the probands from a two-case family was found to have the previously reported Q50R mutation. No family member was found to carry the mutation at position -34 or any other disease-associated mutation. For further investigation of noncoding CDKN2A mutations that may affect transcription, allele specific expression analysis was carried out in 31 of the families with at least three affected members who showed either complete or "indeterminate" 9p haplotype sharing without CDKN2A exonic mutations. Reverse transcription polymerase chain reaction and automated sequencing showed expression of both CDKN2A alleles in all family members tested. The lack of CDKN2A promoter mutations and the absence of transcriptional silencing in the germ line of this cohort of families suggest that mutations in the promoter and 5' UTR play a very limited role in melanoma predisposition. PMID- 11477666 TI - A new gadolinium-based contrast agent for magnetic resonance imaging of brain tumors: kinetic study on a C6 rat glioma model. AB - T1-weighted magnetic resonance imaging (MRI) was used to evaluate the potential interest of a new Gd-based contrast agent, termed P760, to characterize brain tumor heterogeneity and vascularization and to delineate regions containing permeable vessels. The C6 rat glioma model was used as a model of high-grade glioblastoma. The signal enhancement was measured as a function of time in the vascular compartment and in different regions of interest (ROIs) within the tumor after the injection of 0.02 mmol x kg(-1) of P760. The results were compared to those obtained after the injection of 0.1 mmol x kg(-1) of Gd-DOTA. We showed that P760, in spite of a Gd concentration five times smaller, produces an enhancement in the blood pool similar to that produced by Gd-DOTA. It was shown that P760 makes possible an excellent delineation of regions containing vessels with a damaged blood-brain barrier (BBB). Images acquired 5-10 minutes after P760 injection showed the location of permeable vessels more accurately than Gd-DOTA enhanced images. The enhancement produced in the tumor by P760 was, however, less than that produced by Gd-DOTA. The extravasation and/or diffusion rate of P760 in the interstitial medium were found to be strongly reduced, compared to those found with Gd-DOTA. This study suggests that the new contrast agent has promising capabilities in clinical imaging of brain tumors. PMID- 11477667 TI - Aortic and mitral regurgitation: quantification using moving slice velocity mapping. AB - Comprehensive assessment of the severity of valvular insufficiency includes quantification of regurgitant volumes. Previous methods lack reliable slice positioning with respect to the valve and are prone to velocity offsets due to through-plane motion of the valvular plane of the heart. Recently, the moving slice velocity mapping technique was proposed. In this study, the technique was applied for quantification of mitral and aortic regurgitation. Time-efficient navigator-based respiratory artifact suppression was achieved by implementing a prospective k-space reordering scheme in conjunction with slice position correction. Twelve patients with aortic insufficiency and three patients with mitral insufficiency were studied. Aortic regurgitant volumes were calculated from diastolic velocities mapped with a moving slice 5 mm distal to the aortic valve annulus. Mitral regurgitant flow was indirectly assessed by measuring mitral inflow at the level of the mitral annulus and net aortic outflow. Regurgitant fractions, derived from velocity data corrected for through-plane motion, were compared to data without correction for through-plane motion. In patients with mild and moderate aortic regurgitation, regurgitant fractions differed by 60% and 15%, on average, when comparing corrected and uncorrected data, respectively. Differences in severe aortic regurgitation were less (7%). Due to the large orifice area of the mitral valve, differences were still substantial in moderate-to-severe mitral regurgitation (19%). The moving slice velocity mapping technique was successfully applied in patients with aortic and mitral regurgitation. The importance of correction for valvular through-plane motion is demonstrated. PMID- 11477668 TI - Clinical evaluation of aortic diseases using nonenhanced MRA with ECG-triggered 3D half-Fourier FSE. AB - The efficacy of the nonenhanced magnetic resonance angiography (MRA) technique known as fresh-blood imaging (FBI), using electrocardiograph (ECG)-triggered 3D half-Fourier fast spin-echo (FSE), was evaluated for the detection and characterization of aortic diseases. Seventy-five consecutive patients with aortic disease underwent the FBI examination on a 1.5-T clinical imager. The results showed that the FBI technique permits clear visualization of aortic diseases, and the vessel branches and their relationship, which provides valuable information. Therefore, the nonenhanced FBI technique is appropriate to use for screening purposes. PMID- 11477669 TI - In vitro validation of MR measurements of arterial pulse-wave velocity in the presence of reflected waves. AB - A magnetic resonance imaging projective velocity encoding sequence was used to determine the pulse-wave velocity in an artery model. To this end, a well-defined flow phantom simulating flow propagation in large arteries was used. In order to validate the measurement method in the presence of large reflected waves, these were deliberately created in the phantom. The projective sequence was applied to two measurement sites and the wave velocity was determined from the spatial and temporal separations of the foot of the velocity waveform. A theoretical model describing reflection and attenuation phenomena was compared with experimental velocity waveforms. The model showed that reflections and attenuation can explain the important changes in velocity waveforms. The model also confirmed that in the presence of reflecting waves, the foot of the waveform can be used as a characteristic point for measurements through changes in the waveform. PMID- 11477670 TI - Improving the detectability of focal liver lesions on T2-weighted MR images: ultrafast breath-hold or respiratory-triggered thin-section MRI? AB - The purpose of this study was to determine whether a respiratory-triggered (RT) T2-weighted turbo spin-echo (TSE) sequence with thin section can improve the detectability of focal liver lesions compared to a breath-hold (BH) T2-weighted TSE sequence. In 25 patients an RT TSE with 8-mm sections (8-TSE RT) and 5-mm sections (5-TSE RT) and a BH TSE sequence with 8-mm sections (8-TSE BH) were performed. Forty-one focal liver lesions (mean: 1.8 +/- 1.2 cm; 14 lesions < or =1 cm; 27 lesions >1 cm) were evaluated. The 5-TSE RT was significantly better in lesion detection compared to the 8-TSE BH sequence for all sizes of lesions (40/41 vs. 33/41; P = 0.014). For lesions >1 cm no relevant differences in the detection rate of the sequences were found (8-TSE RT, 26/27; 5-TSE RT, 26/27; 8 TSE BH, 25/27), for lesions < or =1 cm the 5-TSE RT provided significantly better sensitivity than the 8-TSE BH (14/14 vs. 8/14, P = 0.015). The results of this study suggest that lesion detection could be significantly improved by using an RT TSE sequence with thin sections compared with a BH TSE sequence. PMID- 11477671 TI - MR renography by semiautomated image analysis: performance in renal transplant recipients. AB - We evaluated a method of semiautomated analysis of dynamic MR image series in renal transplants. Nine patients were studied twice, with an average time interval of 7 days. MR examination consisted of a run of 256 T1-weighted coronal scans (GE; TR/TE/flip: = 11/3.4/60 degrees; slice thickness = 6 mm; temporal resolution = 2 seconds). Gadolinium-DTPA (0.05 mmol/kg) was injected with an injector pump (5 ml/seconds). MR renographs of the cortex and medulla were obtained by segmentation of the renal transplant and placement of two regions of interest (ROIs) overlying the peripheral and central renal parenchyma. In the first 100 frames of the renographs, analysis of variance (ANOVA) demonstrated significant intraclass correlation coefficients with mean values for the cortex and medulla of 0.47 and 0.59, respectively. We conclude that the procedure is a robust technique that generates meaningful signal curves. PMID- 11477672 TI - Extent and time course of morphological changes of bone marrow induced by granulocyte-colony stimulating factor as assessed by magnetic resonance imaging of healthy blood stem cell donors. AB - The aim of this study was to assess the time course and extent of signal alterations of red bone marrow after short-term stimulation by recombinant human granulocyte-colony stimulating factor (rHuG-CSF) in healthy peripheral blood stem cell donors using magnetic resonance imaging (MRI) at low-field strength. Twelve healthy blood stem cell donors without evidence of bone marrow disorders were prospectively investigated and underwent four MRI studies of their lumbar spine. Sagittal T1- and T2-weighted spin-echo sequences and a gradient-echo (GE) sequence with an echo time for out-of-phase imaging were performed prior to rHuG CSF application (baseline MRI), on the day of first stem cell harvest (after 70 microg/kg body weight rHuG-CSF, second MRI) followed by two studies 9-18 days (median 14.5 days, third MRI) and 26-48 days (median 39.5 days, fourth MRI) after discontinuation of rHuG-CSF application. Baseline MRI showed normal marrow signal in all patients. The second MRI revealed a decrease of quantified bone marrow signal relative to nucleus pulposus in T1- and T2-weighted images and an increase of relative signal in out-of-phase GE sequences. The greatest changes of relative marrow signal were observed at the third MRI. Compared to baseline MRI, relative marrow signal was diminished by 12% in T1-weighted images and increased by 59% in GE sequences, consistent with a rise in marrow cellularity simulating diffuse marrow disease. At the fourth MRI quantified relative marrow signal returned to baseline levels in all sequences. In healthy individuals rHuG-CSF application leads to significant signal changes of bone marrow in lumbar vertebra that are maximal about 2 weeks after discontinuation of rHuG-CSF application. In patients with underlying marrow disorders who receive hematopoietic growth factors during treatment, these changes should not be confused with disease progression. PMID- 11477673 TI - Osteodensitometry of human heel bones by MR spin-echo imaging: comparison with MR gradient-echo imaging and quantitative computed tomography. AB - The aim of the study was to investigate whether quantitative magnetic resonance (MR) fast spin-echo (FSE) imaging with moderate spatial resolution enables osteodensitometry in peripheral yellow bone marrow. Signal intensities in T1 weighted FSE images from yellow bone marrow indicate the amount of adipose tissue per volume. The signal intensity in marrow regions with spongy bone was assessed and compared to signal intensity of pure fatty marrow (100%). Heel bones of 30 patients with suspected osteoporosis were analyzed and the FSE images were compared with results from parallel MR gradient-echo (GE) imaging and quantitative computed tomography (QCT) examinations. High correlation was found between FSE imaging and QCT [r = 0.91 in the dorsal region of interest (ROI); r = 0.86 in ventral ROI]. Linear correlation coefficients between GE imaging and QCT were slightly lower in the dorsal part (r = -0.86) and considerably lower in the ventral part (r = -0.68). Correlation between the two MR techniques amounted to r = -0.72/-0.61 (dorsal/ventral). The high correlation between FSE imaging and bone mineral density (BMD) allows possible clinical applications of FSE imaging for diagnosis of osteoporosis. Further improvements of the accuracy using reference phantoms might be possible. PMID- 11477674 TI - BOLD MRI of human tumor oxygenation during carbogen breathing. AB - An MRI method is described for demonstrating improved oxygenation of human tumors and normal tissues during carbogen inhalation (95% O2, 5% CO2). T2*-weighted gradient-echo imaging was performed before, during, and after carbogen breathing in 47 tumor patients and 13 male volunteers. Analysis of artifacts and signal intensity was performed. Thirty-six successful tumor examinations were obtained. Twenty showed significant whole-tumor signal increases (mean 21.0%, range 6.5 82.4%), and one decreased (-26.5 +/- 8.0%). Patterns of signal change were heterogeneous in responding tumors. Five of 13 normal prostate glands (four volunteers and nine patients with nonprostatic tumors) showed significant enhancement (mean 11.4%, range 8.4-14.0%). An increase in brain signal was seen in 11 of 13 assessable patients (mean 8.0 +/- 3.7%, range 5.0-11.7%). T2* weighted tumor MRI during carbogen breathing is possible in humans. High failure rates occurred due to respiratory distress. Significant enhancement was seen in 56%, suggesting improved tissue oxygenation and blood flow, which could identify these patients as more likely to benefit from carbogen radiosensitization. PMID- 11477675 TI - MR lung imaging at 0.2 T with T1-weighted true FISP: native and oxygen-enhanced. AB - An inversion recovery true fast imaging with steady precession (FISP) pulse sequence was developed to carry out fast imaging of the lungs at 0.2 T. Using this sequence, oxygen-enhanced magnetic resonance (MR) lung imaging was performed on healthy volunteers. The lungs showed signal enhancement (11.7% +/- 3.8%) when breathing 100% oxygen. Using inversion recovery, true FISP at low field may prove promising for MR lung imaging. PMID- 11477676 TI - In vivo studies of Gd-DTPA-monoclonal antibody and gd-porphyrins: potential magnetic resonance imaging contrast agents for melanoma. AB - New tumor-specific contrast agents for clinical imaging and therapy for cancer are required. To this end Gd-H (Gd-hematoporphyrin), Gd-TCP (Gd-tetra carboranylmethoxyphenyl-porphyrin), Gd-DTPA-WM53, and Gd-DTPA-9.2.27 were synthesized and administered by systemic injection to nude mice with human melanoma (MM-138) xenografts. The biodistribution T1 relaxation times and magnetic resonance (MR) image signal enhancement of the contrast agents are presented for the first time and compared for each group of five mice. A change (20%) in T1 relaxation times of water in human melanoma tumor xenografts was revealed 24 hours after injection of the labeled immunoconjugate Gd-DTPA-9.2.27. The percent of injected antibody or gadolinium that localized to the tumor was measured by inductively coupled plasma atomic emission spectroscopy (ICP-AES) to be approximately 35%. A higher concentration of gadolinium was achieved compared with nonspecific compounds, indicating selective delivery of Gd-DTPA-9.2.27 to the melanoma xenografts. Porphyrin-based contrast agents (Gd-H and Gd-TCP) also showed significant uptake in melanomas. The uptake of Gd-TCP by the tumor was sufficient to deliver boron atoms into the tumor, making possible dual use for both MR imaging (MRI) and boron neutron capture therapy (BNCT). The linear relationship found between the paramagnetic contribution to the relaxation rates and contrast agent concentration allows quantitative studies of paramagnetic contrast agent uptake. PMID- 11477677 TI - Dynamic observation of pulmonary perfusion using continuous arterial spin labeling in a pig model. AB - The continuous arterial spin-labeling (CASL) method of perfusion MRI is used to observe pulmonary perfusion dynamically in an animal model. Specifically, a respiratory-triggered implementation of the CASL method is used with approximate spatial resolution of 0.9 x 1.8 x 5.0 mm (0.008 cc) and 2-minute temporal resolution. Perfusion MRI is performed dynamically during repeated balloon occlusion of a segmental pulmonary artery, as well as during pharmacological stimulation. A total of three Yorkshire pigs were studied. The results demonstrate the ability of the endogenous spin-labeling method to characterize the dynamic changes in pulmonary perfusion that occur during important physiological alterations. PMID- 11477678 TI - Volume-localized two-dimensional correlated magnetic resonance spectroscopy of human breast cancer. AB - A localized 2D correlation spectroscopic sequence (L-COSY) was implemented and applied in human breast cancer in vivo to evaluate the water to fat (both saturated and unsaturated) ratios and also to identify choline. Being in agreement with the conventional 1D magnetic resonance spectroscopy (MRS) results, elevated water to lipids ratios were found in breast cancers and choline was observed only in a few cancer patients. PMID- 11477679 TI - A low-cost system for monitoring skin conductance during functional MRI. AB - We built a low-cost system for monitoring human skin conductance responses (SCRs) in a clinical magnetic resonance (MR) scanner during functional imaging. The average scanner-induced conductance noise level was suppressed sufficiently to allow SCR measurements over the full range of SCR amplitudes, and functional image signal-to-noise ratio was unaffected by the skin conductance apparatus. The system may be useful for a variety of imaging studies. PMID- 11477680 TI - Evaluation of the anatomical and functional properties of deglutition with various kinetic high-speed MRI sequences. AB - We evaluated various fast MR sequences for obtaining anatomical and dynamic functional information during deglutition. Seven healthy volunteers underwent MRI of the oropharynx during swallowing of an oral positive-contrast agent. Single slice imaging was performed in the median sagittal plane while subjects were in a supine position. Twenty serial images were obtained using EPI, FLASH, and turbo FLASH sequences. The dynamic (movement-related) information and the anatomical resolution of the soft tissues were evaluated during deglutition. The FLASH sequence provided high-quality images at rest. During swallowing, however, the images were significantly degraded by movement artifacts and had inferior temporal resolution. The EPI evidenced better temporal resolution, but was degraded by strong distortions and movement artifacts. The turbo-FLASH sequence provided the best temporal resolution and sufficient spatial resolution during motion. This sequence proved optimal for the investigation of swallowing function, and is expected to be of value for the documentation of functional disturbances in patients with oropharyngeal pathology. PMID- 11477681 TI - Morphant technology in model developmental systems. PMID- 11477682 TI - Achieving efficient delivery of morpholino oligos in cultured cells. AB - One of the many features that make morpholino oligos unique among the antisense structural types is an uncharged backbone. While this feature eliminates the nonspecific interactions of traditional S-oligos, it also renders the morpholino undeliverable via the traditional lipid-based delivery systems. This article describes a highly efficient method of delivering morpholino oligos into adherent and nonadherent cultured cells. In this system, a nonionic morpholino oligo is paired to a complementary DNA "carrier." The DNA is then bound electrostatically to a partially ionized, weakly-basic ethoxylated polyethylenimine (EPEI). This morpholino/DNA/EPEI complex is efficiently endocytosed, and when the pH drops within the endosome, the EPEI more fully ionizes, resulting in permeabilization of the endosomal membrane and release of the morpholino into the cytosol. This article describes optimization of delivery in HeLa cells and provides the basis for delivery in any cultured endocytic cell type. genesis 30:94--102, 2001. PMID- 11477683 TI - Action of morpholinos in Ciona embryos. AB - First, using morpholino against lacZ, we demonstrate that the morpholino specifically suppresses the translation of the gene introduced exogenously into Ciona eggs. Second, using morpholino against an alkaline phosphatase gene, we show that the morpholino suppresses the translation of the endogenous gene as well. Third, using morpholino against beta-catenin gene, we confirm that the suppression by the morpholino can be rescued by injection of beta-catenin mRNA. All of these results indicate that morpholino act in Ciona embryos to specifically block the function of endogenous genes as well as exogenously introduced genes. genesis 30: 103--106, 2001. PMID- 11477684 TI - Cyclin E morpholino delays embryogenesis in Xenopus. PMID- 11477685 TI - Comparison of morpholino based translational inhibition during the development of Xenopus laevis and Xenopus tropicalis. AB - Morpholino (MO) based inhibition of translational initiation represents an attractive methodology to eliminate gene function during Xenopus development (Heasman et al., 2000). However, the degree to which a given target protein can be eliminated and the longevity of this effect during embryogenesis has not been documented. To examine the efficacy of MOs, we have used transgenic Xenopus lines that harbour known numbers of integrations of a GFP reporter under the control of the ubiquitous and highly expressed CMV promoter (Fig. 1a). In addition we have investigated the longevity of the inhibitory effect by using transgenic lines expressing GFP specifically in the lens of tadpoles. These transgenic lines represent the ideal control for the technique as the promoters are highly expressed and GFP can be easily detected by fluorescence and immunoblotting. Moreover, as GFP has no function in development, the levels of inhibition can be tested in an otherwise normal individual. Here we report that MOs are able to efficiently and specifically inhibit the translation of GFP in transgenic lines from Xenopus laevis and Xenopus tropicalis and the inhibitory effect is long lived, lasting into the tadpole stages. genesis 30:110--113, 2001. PMID- 11477686 TI - Zebrafish frizzled-2 morphant displays defects in body axis elongation. PMID- 11477687 TI - Xenopus frizzled-7 morphant displays defects in dorsoventral patterning and convergent extension movements during gastrulation. PMID- 11477688 TI - Target selection for Danio rerio functional genomics. PMID- 11477689 TI - A morpholino phenocopy of the cyclops mutation. PMID- 11477690 TI - Morpholino-induced knockdown of zebrafish engrailed genes eng2 and eng3 reveals redundant and unique functions in midbrain--hindbrain boundary development. PMID- 11477691 TI - Pdx-1 knockdown reduces insulin promoter activity in zebrafish. PMID- 11477692 TI - Zebrafish pdx1 morphant displays defects in pancreas development and digestive organ chirality, and potentially identifies a multipotent pancreas progenitor cell. PMID- 11477693 TI - Zebrafish hhex regulates liver development and digestive organ chirality. PMID- 11477694 TI - Pitx1 and Pitx2c are required for ectopic cement gland formation in Xenopus laevis. AB - The mucus secreting cement gland is the anterior-most ectodermal organ of the Xenopus embryo. The homeobox genes Pltx1 and Pitx2c are expressed in the cement gland primordium. Misexpression of both genes induced ectopic cement gland tissue in whole embryos and transcription of the marker genes Xag1 and Xag2 in animal cap explant cultures. Antisense morpholino oligonucleotides against Pitx1 and Pitx2c inhibited ectopic cement gland formation induced by otx2. Gene knock downs generated by morpholino oligonucleotides were specific and could be rescued by coinjection of Pitx mRNAs. These data demonstrate for the first time the requirement of specific genes for cement gland formation by loss-of-function experiments. genesis 30:144--148, 2001. PMID- 11477695 TI - Inhibition of skiA and skiB gene expression ventralizes zebrafish embryos. PMID- 11477696 TI - Inhibition of zebrafish fgf8 pre-mRNA splicing with morpholino oligos: a quantifiable method for gene knockdown. PMID- 11477697 TI - Morpholino-induced knockdown of fgf8 efficiently phenocopies the acerebellar (ace) phenotype. PMID- 11477698 TI - Morpholino phenocopies of the bmp2b/swirl and bmp7/snailhouse mutations. PMID- 11477699 TI - Floor plate develops upon depletion of tiggy-winkle and sonic hedgehog. PMID- 11477701 TI - Morpholino phenocopies of sqt, oep, and ntl mutations. PMID- 11477700 TI - Sonic hedgehog and tiggy-winkle hedgehog cooperatively induce zebrafish branchiomotor neurons. PMID- 11477702 TI - Morpholino knock-down of antivin1 and antivin2 upregulates nodal signaling. PMID- 11477703 TI - A zebrafish vasa morphant abolishes vasa protein but does not affect the establishment of the germline. PMID- 11477704 TI - Morpholino phenocopies of endothelin 1 (sucker) and other anterior arch class mutations. PMID- 11477705 TI - A morpholino phenocopy of the colourless mutant. PMID- 11477706 TI - Morpholino phenocopies of the swirl, snailhouse, somitabun, minifin, silberblick, and pipetail mutations. PMID- 11477707 TI - Suppression of heat shock transcription factor HSF1 in zebrafish causes heat induced apoptosis. PMID- 11477708 TI - A morpholino phenocopy of the mouse mos mutation. PMID- 11477709 TI - Extraction of DNA from oral cytological samples by scraping and smear method suitable for restriction site mutation analysis: a pilot study. AB - The restriction site mutation assay (RSM) can be used to measure base changes which occur in the DNA coding for bacterial restriction enzymes. The aim of this study was to investigate whether DNA, of sufficient quantity and quality for analysis by RSM, could be extracted from cells collected from precancerous lesions using a cytological smear technique. Six smears were collected from each of five lesions of leukoplakia displaying a variety of clinical appearances. Three methods for the extraction of DNA were compared. The commercial extraction method was shown to be most convenient and reproducible, routinely providing 1-5 microg of DNA per sample. Cell populations collected by a cytological smear technique can provide DNA of sufficient quantity and quality for analysis of RSM. PMID- 11477710 TI - Follicular variant of papillary carcinoma: cytologic findings on FNAB samples experience with 16 cases. AB - Between January 1, 1992 and December 31, 1997, a cytopathological diagnosis of follicular variant of papillary thyroid carcinoma (FVPC) was made on a series of 16 out of 18 patients with palpable nodules who underwent fine-needle aspiration biopsy (FNAB) in our Department. The results of aspiration biopsy were followed by histopathological examination of the surgically excised tissues. There were three false-negative aspirations (16.6%), of which two were probably bound to fine-needle sampling and one due to a mixture of benign and malignant cells which had originally gone unrecognized. The accuracy of the cytopathologic diagnosis in this variant was 88.8%. An analysis of the diagnostic cytopathological criteria was performed, which demonstrated the importance of both architectural features (monolayered and branching sheets, microacinar structures, and their combinations) and nuclear features (presence of nuclear grooves). Background bound features were mainly represented by dense, nonfilamentous colloid. The cytopathologic findings in FVPC were compared to those found in a series of 10 usual papillary carcinomas (UPC) and 10 follicular neoplasms (FN). These latter had originally been diagnosed by FNAB and were subsequently classified histologically as follicular adenoma (n = 6), follicular carcinoma (n = 3), or adenomatoid colloid nodule (n = 1). Statistical evaluation was performed on the cytopathological findings in the three classes of lesions (FVPC, UPC, and FN) as to their presence and relative frequency or absence by using a nonparametric one way ANOVA (Kruskall-Wallis) and, where necessary, a Mann-Whitney U test. Papillary cellular fragments and multinucleated giant cells (P < 0.005), nonfilamentous dense colloid, squamoid cells, and syncytia were significantly more represented in UPC than in FVPC (P < 0.05), while histiocytes were significantly more frequent in FVPC (P < 0.005). Other nuclear and/or background features were significant only in the distinction between papillary carcinomas as a group and FN. The cytological differential diagnosis of the FVPC is briefly discussed with relevance to the possible pitfalls caused by its peculiar cyto- and histomorphology. PMID- 11477711 TI - Spindle-cell lesions of the liver: diagnosis by fine-needle aspiration biopsy. AB - Rarely, spindle-cell lesions in liver fine-needle aspiration biopsies (FNABs) are encountered. A retrospective review of our experience with lesions that are mesenchymal in origin or appearance was undertaken to elucidate the frequency and spectrum of these lesions. Image-guided liver FNABs performed over a 3-year period (n = 585) at our institution (1996-1998) were retrospectively evaluated. Cytologic smears, cell block preparations, and clinical follow-up of lesions with spindle-cell morphology were reviewed. Twenty-nine of 585 cases were of spindle cell morphology (5%). Hemangiomas (n = 12, 41%) and metastatic sarcomas (n = 6, 21%) comprised the largest categories, followed by granulomatous inflammation (n = 3, 10%). Other cases included primary angiosarcoma and fibrolamellar hepatocellular carcinoma. The most frequent spindle-cell liver lesion encountered is hemangioma, followed by metastatic leiomyosarcoma and granulomatous hepatitis. Awareness of diagnostic possibilities, special attention to specimen adequacy, and use of ancillary procedures can maximize diagnostic yield. PMID- 11477712 TI - Routine use of ThinPrep method in fine-needle aspiration material as an adjunct to standard smears. AB - The purpose of this study was to evaluate the efficacy of routine use of ThinPrep in fine-needle aspiration (FNA) material as an adjunct to standard smears. We reviewed 50 consecutive, satisfactory-for-evaluation FNA materials in which ThinPrep slides were obtained. In each case, Diff-Quik-stained and Papanicolaou stained smears were initially prepared, and the ThinPrep slide was made from the needle/syringe rinse. Smears and ThinPrep slides were evaluated for the presence of diagnostic material. In addition, the following questions were addressed: could the final diagnosis be made based on smears only, and did the ThinPrep method add any additional information to the diagnosis? Smears were satisfactory for evaluation in 49/50 cases, and in one case diagnostic material was present on the ThinPrep slide only. Thirty-two out of 50 ThinPrep slides contained material sufficient for evaluation. In 46/50 (92%) cases, final diagnoses could be made based on smears only. In 2 cases, ThinPrep slides showed additional diagnostic material, and in another 2 cases were critical for the diagnosis. The routine use of ThinPrep as an adjunct preparatory method to FNA material is not justified, and is associated with an additional, potentially nonreimbursable cost. PMID- 11477713 TI - Clinical implications and value of immunohistochemical staining in the evaluation of lymph node infarction after fine-needle aspiration. AB - We report on a series of 3 patients who underwent fine-needle aspiration (FNA) for clinically apparent lymphadenopathy. In all 3 cases, a diagnosis of malignancy was rendered based on cytologic findings (two metastatic squamous-cell carcinomas and one melanoma). However, initial follow-up surgical pathology reported only "extensive coagulative necrosis, no viable tumor seen." Subsequent immunohistochemical stains (cytokeratins (AE1/AE3), HMB45, S100, and Melan A) demonstrated the presence of metastatic tumor in the area of infarction in each case, thus establishing the presence of metastatic tumor and correct interpretation of the initial FNA. We conclude, based on our own experience and a few previously reported cases, that total infarction of the lymph nodes following FNA can occur, and immunohistochemistry can be helpful in clinical management. PMID- 11477714 TI - Primary malignant melanoma of the uterine cervix: report of a case diagnosed by cervical scrape cytology and review of the literature. AB - Cervical melanoma is a rare neoplasm and is seldom diagnosed by cervical scrape cytology. The possibility of metastatic melanoma should be excluded before making a diagnosis of primary cervical melanoma. We present a case of primary cervical melanoma diagnosed by cervical scrape cytology and a review of literature. The patient presented with vaginal bleeding of 3 mo duration and an ulcerated cervical growth. Cervical smear showed bizarre and abnormal cells containing pigment. PMID- 11477715 TI - Lymph node cytology in Wegener's granulomatosis. AB - We report on cytological findings on aspirates from the cervical lymph node from a case of Wegener's granulomatosis (WG). The diagnostic utility of technique in diagnosing a sizable minority of WG patients who present with cervical lymphadenopathy is discussed. We outline an approach to diagnosis of necrotising granulomatous lesions in aspirates from lymph nodes in a tabular form. PMID- 11477716 TI - Phyllodes tumor of the breast with actin inclusions in stromal cells: diagnosis by fine-needle aspiration cytology. AB - The occurrence of hyaline inclusions in stromal cells in fibroepithelial tumors of the breast is very uncommon. These inclusions, characteristic of infantile digital fibromatosis, are comprised of actin filaments. This report illustrates a case of a benign phyllodes tumor of the breast with inclusion bodies, identified by fine-needle aspiration. Histologically, many of the stromal cells contained round intracytoplasmic inclusions, with positivity for smooth muscle actin. PMID- 11477717 TI - Fine-needle aspiration cytology of pancreatoblastoma. AB - Pancreatoblastoma is a rare pancreatic neoplasm seen most commonly in the pediatric age group. We report on the aspiration cytology and immunohistochemical findings of a pancreatoblastoma in a 16-yr-old male. PMID- 11477718 TI - Fine-needle aspiration cytology of "proximal-type" epithelioid sarcoma. AB - The cytologic and immunocytologic findings in a case of recurrent "proximal-type" epithelioid sarcoma (ES) of the vulva are presented. This is a recently described neoplasm that differs clinically and morphologically from conventional ES. Cytologic smears showed a dissociated population of large, atypical neoplastic cells with bi- and multinucleated cells, abundant cytoplasm, and rhabdoid-like morphology. Due to its different clinical management it must be differentiated from metastatic carcinoma and melanoma. From a practical perspective, its differentiation from other epithelial-like sarcomas is less important. In conclusion the cytopathologic findings of "proximal-type" ES show a good correlation with histopathology, permitting the diagnosis of recurrences and metastases. When accompanied by adequate clinical information and ancillary studies, a specific preoperative recognition seems possible. PMID- 11477719 TI - Fine-needle aspiration cytology of small-cell carcinoma of the parotid. AB - Small-cell carcinomas arise uncommonly in extrapulmonary sites and are rare primary neoplasms in the salivary glands. We report on the aspiration cytology and immunohistochemical findings of a small-cell carcinoma of the parotid gland in an 81-yr-old man. PMID- 11477720 TI - Lipid-rich carcinoid tumor of the thymus gland: diagnosis by fine-needle aspiration biopsy. AB - In this report we describe the cytologic features of an unusual thymic carcinoid tumor containing prominent cytoplasmic lipid vacuoles that was diagnosed by fine needle aspiration biopsy. The tumor presented as a presternal subcutaneous mass in an 81-yr-old female, with a contiguous mediastinal mass on computed tomography. The cytomorphologic features included numerous discohesive cells with eccentric, round to oval nuclei, granular chromatin, and scant cytoplasm containing numerous intracytoplasmic, clear vacuoles. The neoplastic cells were reactive for neuroendocrine markers by immunocytochemistry and showed reactivity of the intracytoplasmic vacuoles with an oil red-O stain for neutral lipid. Ultrastructural examination confirmed the presence of intracytoplasmic lipid vacuoles and neurosecretory granules. Subsequent surgical excision confirmed the diagnosis. We believe this to be the first report describing these features in a primary thymic carcinoid tumor. The differential diagnosis of mediastinal tumors with clear-cell features and cytoplasmic vacuolization is also discussed. PMID- 11477721 TI - Cytologic features of disseminated bacillus Calmette-Guerin (BCG) infection. AB - Disseminated bacillus Calmette-Guerin (BCG) infection is an unusual complication of immunization against Mycobacterium tuberculosis with the bacillus Calmette Guerin. We report on 4 such cases in which the diagnosis was suspected at the fine-needle aspiration biopsy (FNAB) procedure. Participants were 4 males (mean age, 21.5 mo; range, 8-36 mo) in good general condition, in whom epidemiology data favoring tuberculosis and presence of pulmonary tuberculosis were lacking. Cases 1 and 2 presented with a deep-seated subcutaneous nodule located near the left mamilla and lower aspect of the left scapula, respectively, resulting from lymph node involvement by BCG. Cases 3 and 4 presented as an osteolytic lesion of the ninth right rib and right iliac bone, respectively. FNAB findings showed poorly to moderately cellular smears. Epithelioid histiocytes in a granuloma pattern with occasional multinucleated Langerhans-type giant cells, lymphocytes, and polymorphonuclear leukocytes in a finely granular background with necrotic debris were found in all cases. The presence of isolated calcified spherules interspersed among the cells was found to be a useful finding for diagnosis. When dealing with disseminated BCG infections, clinical and cytological pictures must be evaluated as a whole in order to arrive at an accurate diagnosis. PMID- 11477722 TI - Smears diagnosed as ASCUS: interobserver variation and follow-up. AB - The purpose of this study was to apply atypical squamous cells of undetermined significance (ASCUS) criteria from the Bethesda System for Reporting Cervical/Vaginal Cytologic Diagnoses (TBS) to the rescreen of cases previously diagnosed as ASCUS, to compare initial and rescreen diagnoses, and to analyze agreement with follow-up (cytology or histology). Two cytotechnologists (S.B. and M.J.M.) and one cytopathology fellow (M.A.) rescreened 632 cervicovaginal specimens diagnosed as ASCUS between June 1, 1992-December 31, 1995. Age and LMP were provided. Rescreen diagnoses were categorized as within normal limits (WNL), ASCUS, low-grade squamous intraepithelial lesions (LSIL), high-grade squamous intraepithelial lesions (HSIL), or carcinoma (CA). Complete agreement was found in 200 specimens (32%): 31 (15%) WNL; 91 (45%) ASCUS; 77 (38.5%) SIL; and one (0.50%) CA. Follow-up revealed no abnormality in 67% of the cases reclassified as WNL, 49% of the cases reclassified as ASCUS, and 48% of the cases reclassified as squamous intraepithelial lesions (SIL). SIL was found in 29% of cases reclassified as WNL, 29% of specimens rediagnosed as ASCUS, and 34% of cases reclassified as SIL. Partial agreement was found in 391 specimens (62%). In 41 specimens (6%), rescreeners were in complete disagreement, and follow-up revealed 9/41 (22%) SIL or worse; 21/41 (51%) WNL; and 4/41 (10%) inconclusive. Applying established criteria, 14% (91/632) of cases diagnosed as ASCUS resulted in complete agreement, and 30% (190/632) resulted in partial agreement. Follow-up of cases initially diagnosed as ASCUS revealed SIL or CA in 30% of cases. ASCUS is a significant diagnosis warranting careful patient follow-up. PMID- 11477723 TI - Utility of additional slides from residual Preservcyt material in difficult ThinPrep gynecologic specimens: a prospective study of 58 cases. AB - ThinPrep purportedly increases the sensitivity of cervicovaginal cytology for detecting abnormal squamous and glandular cells. The value of additional slides from residual Preservcyt material to characterize difficult lesions is unknown. Fifty-eight cases were studied to determine the utility of additional slides for diagnosis and to assess cellular uniformity. In 32 (55%), repeat slides helped make a definitive diagnosis, including 18 atypical squamous cells of uncertain significance (ASCUS) reclassified as low-grade squamous intraepithelial lesion (LGSIL) (13), high-grade squamous intraepithelial lesion (HGSIL) (4), or endometrial adenocarcinoma (1); 5 LGSIL reclassified as HGSIL; 3 atypical glandular cells of uncertain significance (AGUS) reclassified as LGSIL (1) or HGSIL (2); 2 LGSIL?HGSIL classified as LGSIL; and 4 cases confirmed as LGSIL (2) or HGSIL (2). Results were compared to follow-up clinical information, including subsequent cervicovaginal samples and biopsies. The number of abnormal cells was similar between slides in most cases. We conclude that, while ThinPreps prepared from the same vial have similar numbers of abnormal cells, additional slides can be helpful for diagnosis in select cases. PMID- 11477724 TI - Previous reports of histiocytic proliferative disorders. PMID- 11477726 TI - Prospective study of the patient-level cost of asthma care in children. AB - Our objective was to assess the cost of asthma care at the patient level in children from the perspectives of society, the Ontario Ministry of Health, and the patient. In this longitudinal evaluation, health service use data and costs were collected during telephone interviews at 1, 3, and 6 months with parents of 339 Ontario children with asthma. Direct costs were respiratory-related visits to healthcare providers, emergency rooms, hospital admissions, pulmonary function tests, prescription medications, devices, and out-of-pocket expenses. Indirect costs were parents' absences from work/usual activities and travel and waiting time. Hospital admissions accounted for 43%, medications for 31%, and parent productivity losses for 12% of total costs from a societal perspective. Statistically significant predictors of higher total costs were worse symptoms, younger age group, and season of participation. Adjusted annual societal costs per patient in 1995 Canadian dollars varied from $1,122 in children aged 4-14 years to $1,386 in children under 4 years of age. From the Ministry of Health perspective, adjusted annual costs per patient were $663 in children over 4 years and $904 in younger children. Adjusted annual costs from the patient perspective were $132 in children over 4 years and $129 in children under 4 years. The rising incidence of pediatric asthma demands that greater attention be paid to the delivery of optimal care to this segment of the population. Appropriate methods must be used to analyze healthcare costs and the use of services in the midst of widespread healthcare reform. The quality of clinical and health policy decision making may be enhanced by cost-of-illness estimates that are comprehensive, precise, and expressed from multiple perspectives. PMID- 11477727 TI - How pediatricians manage asthma in Thailand. AB - Currently, there is no existing information regarding prescribing practices for the management of childhood asthma among pediatricians in Thailand. In order to evaluate the management standards for childhood asthma in Thailand, 400 self administered questionnaires were randomly mailed to nonacademic pediatricians throughout Thailand, asking questions about their preferences in the treatment of childhood asthma. One hundred and seventy-four of these 400 questionnaires were returned (a response rate of 43.5%). Data were analyzed using the descriptive module of the Epi-info 6 program. For acute asthma, 17% of the respondents used objective measures such as peak flow meters in assessing asthma severity and severity of acute asthma attacks. The drug of first choice for treating acute attacks was a nebulized beta-agonist q 20 min (81.8%). Although 93% indicated that they had used theophylline for treating acute attacks, most would reserve the drug for patients with severe symptoms. Corticosteroids were reserved for those with severe attacks (91.7% both for clinic and for in-hospital settings). Hydrocortisone was the most preferred corticosteroid preparation (59.8%). Ninety seven percent used antibiotics in treating acute asthma, but only with appropriate indications. For chronic asthma, a strong preference was observed for oral beta-agonists as the bronchodilator of choice (88%). For moderately severe asthmatics, theophylline was still preferred by 41% of the responders. Among prophylactic agents, ketotifen was the most favored drug (90.4%), whereas inhaled steroids and cromolyn were chosen by 9.6% and 2.4%, respectively. Eighty-five percent indicated that they would prescribe prophylactic agents for 1 year or less. Forty-two percent never considered allergy evaluation as a part of a workup for childhood asthma. Certain prescribing practices of childhood asthma management in Thailand were observed among pediatricians, i.e., 1) low frequency of using objective measures in assessing asthma severity among pediatricians; 2) frequent use of theophylline and antibiotics in the treatment of acute asthma; 3) late introduction of corticosteroids in treating acute asthma; 4) preference for oral bronchodilators; and 5) preference of ketotifen as the prophylactic drug of choice. This survey provides baseline data and will aid in the evaluation of management guidelines for childhood asthma in Thailand. PMID- 11477728 TI - Effects of single-dose fluticasone on exercise-induced asthma in asthmatic children: a pilot study. AB - A single high dose of inhaled corticosteroid (ICS) can increase airway caliber in children with asthma attacks and laryngitis subglottica. Presumably the effect is due to the vasoconstrictive and antiedematous properties of topical steroids. Enlarged vessels have been suggested to play a role in the pathophysiology of exercise-induced bronchial obstruction (EIB). To investigate this, we evaluated the effect of a single high dose of fluticasone propionate (FP) on EIB in asthmatic children. Nine children aged 8-16 years with mild to moderate asthma were included. All children had a history of EIB, which was confirmed by an exercise test. None was taking ICS maintenance therapy. The children inhaled either a single dose of 1 mg FP or placebo on 2 separate days within 7-14 days. After inhalation, airway caliber (FEV(1)) was assessed for 4 hr before exercise. Then an exercise challenge was performed on a treadmill to assess EIB (% fall FEV(1)). A significant increase in FEV(1) was observed 1 hr after inhalation of FP compared to placebo. Response to exercise was expressed as maximal % fall in FEV(1) from baseline (% fall) and as area under the curve (AUC) of the 30-min time/response curve. The % fall FEV(1) after exercise and the AUC were significantly reduced when FP was inhaled compared to placebo inhalation (% fall 9.7% vs. 19.2%, respectively, P = 0.038 and AUC 92.0%.min vs. 205.7%.min, respectively, P = 0.03). There was considerable individual variability in reduction of EIB, with 5 out of 9 children having a clinically significant response. We conclude that a single high dose of inhaled FP has an acute protective effect on the bronchial response to exercise in a substantial proportion of asthmatic children. PMID- 11477729 TI - Safety of aerosolized INS 365 in patients with mild to moderate cystic fibrosis: results of a phase I multi-center study. AB - Cystic fibrosis (CF) is characterized by defective cystic fibrosis transmembrane regulator (CFTR) expression and function, associated with abnormal ion transport and mucociliary clearance, and clinical lung disease. Triphosphate nucleotides such as uridine-5'-triphosphate (UTP) and INS 365, may be useful for CF through actions, mediated via P2Y(2) extracellular receptors, on chloride and liquid secretion, and ciliary beat frequency. INS 365 may offer chemical stability advantages over UTP. In a randomized, double-blind, multicenter phase I study, we studied the safety and maximally tolerated dose of escalating, single doses of aerosolized INS 365, in adult and pediatric patients with mild to moderate CF lung disease (FEV(1) > or = 45% predicted). In four successive dose cohorts of adult patients (n = 12 per cohort, age > or = 18 years) and four successive pediatric dose cohorts (n = 12 per cohort, age 5-12 years), patients were randomized 3:1 active/placebo (0.9% saline) to evaluate doses of 20, 40, 80, and 100 mg INS 365 delivered by nebulizer (Pari Star ). Sputum was collected pre- and post-dosing to obtain preliminary results on clinical efficacy. After each dose cohort, a Data Safety Monitoring Committee (DSMC) reviewed the data. Forty-eight adult and 36 pediatric patients completed the protocol (up to 100 mg for adults, 80 mg for pediatric patients). The predominant adverse events were cough, wheezing, chest tightness, and a decrease in FEV(1) (occurring in 8/48 adults, and 5/36 pediatric patients), which occurred predominantly in the 80-mg and 100 mg dose cohorts. Though a few adult patients had a tendency to increase sputum production, there was little consistent effect noted on sputum production in this acute, single-dose study. The data suggest that aerosolized INS 365 is safe when delivered at single doses of up to 40 mg in adults and children with CF, but that higher doses are unlikely to be tolerated. PMID- 11477730 TI - Down-regulation of regulatory proteins for differentiation and proliferation in murine fetal hypoplastic lungs: altered mesenchymal-epithelial interactions. AB - We compared proliferation (growth) and differentiation (development) related proteins in normal and hypoplastic fetal murine lungs. The hypoplastic lungs were created in CD-1 fetal mice by nitrofen exposure (25 mg per pregnant mouse given intragastrically on gestational day 8 [Gd8]), as published earlier. The lungs were harvested at Gd14, 16, 19 and from neonates. Immunoblot analyses were carried out for transcription factors (oncogenic proteins, nuclear receptor, and transmembrane receptor proteins) in severely hypoplastic murine fetal lungs with coexistent diaphragmatic hernia, and results were compared with those derived from normal lungs of equivalent age. These proteins have proposed roles in the regulation of proliferation and differentiation processes of fetal lungs. We have shown that the product of the oncogene c-myc was reduced in hypoplastic lungs at all stages of gestation, whereas c-Fos protein levels were variable. These proteins are known to regulate transcription of various developmental proteins, such as those responsible for proliferation and differentiation. Further, the nuclear transcription factors thyroid transcription factor-1 (TITF-1) and glucocorticoid receptor (GR) were reduced, and thyroid hormone receptor (TR) and retinoic acid receptors (RARs) were inhibited in severely hypoplastic lungs compared to normal lungs of equivalent gestational stage, except in neonatal lungs, where signals for RARs were seen. TITF-1 is known to localize in bronchial epithelial cells in developing lungs. It is restricted to type II pneumocytes with gestational development in the normal lungs and regulates surfactant proteins. Earlier, we have reported that surfactant proteins are reduced in hypoplastic lungs. In the current study, reduced GR and TITF-1 proteins may play a role in reducing surfactant proteins in the hypoplastic lungs. The significant inhibition in TR and RARalpha in the severely hypoplastic lungs reflects on affected epithelial cell maturation and alveolar formation, respectively. Altered RARbeta levels correlate with affected lung growth and branching morphogenesis of nitrofen-exposed lungs. A transmembrane receptor protein EGFR was reduced in hypoplastic lungs, suggesting the involvement of altered mesenchymal-epithelial signal transduction pathways. We conclude (1) Our data suggest altered levels of various nuclear transcription factors in the murine fetal hypoplastic lungs; (2) Reduced levels TITF-1 protein in hypoplastic lungs may have caused the functional immaturity of distal lung, immature airways and thus may affect overall differentiation of lungs. These results correlated with low levels of surfactant proteins in these lungs; (3) TR and RAR inhibition indicate their roles through reduced or retarded proliferation and differentiation processes in the severely hypoplastic lungs; (4) GR down-regulation in developing fetal murine hypoplastic lungs indicate delayed development, and GR up-regulation in affected neonates may be induced by stress/stretch caused at birth due to air-breathing; (5) Down- regulation of EGFR indicate altered mesenchymal-epithelial interactions and possible influence on lung proliferation and differentiation. PMID- 11477731 TI - Intratracheal administration of perfluorochemical-gentamicin suspension: a comparison to intravenous administration in normal and injured lungs. AB - Respiratory infections can lead to acute lung injury and perfusion abnormalities. We hypothesized that intratracheal (IT) administration of a perfluorochemical (PFC) gentamicin (G) suspension as compared to intravenous (IV) administration of gentamicin will result in higher lung tissue levels of gentamicin, while maintaining safe serum levels. To test this hypothesis, 21 lambs with normal and acid injured lungs were studied for 4 hr, using 2 different drug delivery methods, IT and IV. Lungs were injured with warm HCl acid in saline lavage, followed by partial liquid ventilation with perflubron (bolus FRC = 20 mL/kg). G at a dose of 5 mg/kg was delivered either IT (G-PFC; 20 mL/kg) or IV (aqueous injection with IT 20 mL/kg PFC alone). Throughout the study, serum G levels, arterial blood gases, respiratory system compliance, and mean arterial blood pressure were measured. Lung tissue G levels were measured at 4 hr and averaged across lobes. Physiologic gas exchange and pulmonary function were maintained throughout the protocol for both the normal and injured lungs. Intravenously administered G resulted in an initial 5-min serum concentration of 43 +/- 2.5 mcg/mL, followed by an exponential decline over the 4-hr protocol to a level of 2.1 +/- 0.23 mcg/mL at hr 4. The intratracheally administered G suspension resulted in a 5-min serum concentration of 1.8 +/- 0.98 mcg/mL and remained relatively constant throughout the protocol, with a 4-hr level of 1.6 +/- 0.29 mcg/mL. With respect to lung tissue G levels, IT administration was significantly more effective in delivering the drug to the normal lungs than IV (31.4 +/- 3.3 mcg/g vs. 4.0 +/- 0.7 mcg/g) 4 hr after administration. In the lung injury group, there was a small but significant difference in lung tissue G levels, with the IT administered perfluorochemical-G suspension achieving greater levels than the IV administered G (11.9 +/- 0.52 mcg/g vs. 10.1 +/- 0.8 mcg/g). Additionally, the drug delivered IV and IT in both the normal and injured lung models was homogeneously distributed throughout the lung. These data show that G lung tissue levels in both normal and injured lungs were higher in the IT group when compared to IV administration. The results of this study demonstrate that in normal and injured lungs, homogeneous G lung tissue levels can be more effectively achieved at lower serum levels when delivered IT in a G-PFC suspension as compared to IV administration. PMID- 11477732 TI - Sputum processing for evaluation of inflammatory mediators. AB - Neutrophil-dominated inflammation is prominent in the cystic fibrosis (CF) and chronic bronchitis (CB) airways. We assessed the degree of airway inflammation by measuring the sputum concentrations of interleukin (IL)-8, myeloperoxidase (MPO), and deoxyribonucleic acid (DNA). We determined the relationship among the concentrations of these mediators and investigated methodological problems that may be responsible for reported variability in measurements. Sputa obtained from 31 patients were solubilized with phosphate-buffered saline, dithiothreitol (DTT) (0.1% or 1%), or dornase alfa (0.2 mg/mL). The sputum concentration of IL-8 and MPO was measured by enzyme-linked immunosorbent assay (ELISA), and DNA was measured using microfluorimetry. There was a significant relationship among sputum IL-8, MPO, and DNA. For MPO (means +/- SD), CF was 1,392 +/- 771 vs. CB at 75 +/- 65 mcg/mL; P < 0.0001. For IL-8: CF was 239 +/- 154 vs. CB at 121 +/- 108 ng/mL; P = 0.0002. For DNA, CF was 1.707 +/- 1.25 vs. CB at 0.184 +/- 0.272 mg/mL; P < 0.0001. The MPO concentration in CF sputum was approximately double after in vitro treatment with dornase alfa (P < 0.0001). There is a greater concentration of IL-8, MPO, and DNA in CF than in CB sputa. There is a significant relationship among these inflammatory markers in sputum. DNA polymers bind myeloperoxidase in the sputum, and we speculate that treatment with dornase alfa may remove a source of MPO inhibition. PMID- 11477733 TI - Off-line exhaled nitric oxide measurements in children. AB - The concentration of exhaled nitric oxide (eNO) is a useful marker of asthmatic bronchial inflammation. eNO can now be measured away from the laboratory (off line), even in children. Short exhalation maneuvers (8 sec) and small samples (1 L) of exhaled gas are probably sufficient in children, but more information is needed about the effect of different measurement conditions. As a preliminary step before conducting epidemiological studies in schoolchildren, we investigated the effects of expiratory flow, dead space, and expiratory time on eNO concentrations collected in 1-L mylar collection bags. We studied 101 cooperative subjects (62 males) aged 5-18 years (30 healthy volunteers, 51 asthmatics, and 20 children with various other respiratory diseases) in our pulmonary function laboratory. On-line and off-line eNO were compared in a single session, and analyzed with a Sievers NOA 280 nitric oxide analyzer. For both methods of collecting expired gas, subjects did a single exhalation without breath-holding against an expiratory pressure 10 cm H(2)O. We investigated the effects of expiratory flow, dead space, and exhalation time on eNO; we also compared on-line and off-line eNO measurements, and the repeatability of both techniques at a given flow rate. Expiratory flows of 58 mL/sec provided more reproducible data than lower flows (coefficient of repeatability 1.1 ppb for 58 mL/sec vs. 2.8 for 27 mL/sec vs. 5.7 for 18 mL/sec). eNO concentrations were about 25% higher in off line than in on-line recordings if the initial 250 mL of exhaled gas were not eliminated, and 37% higher if exhalation lasted longer (16 sec vs. 8 sec). Eliminating 250 mL of dead space and shortening the filling time to 8 sec yielded off-line eNO values close to those on-line (geometric mean off-line eNO 14.4 ppb, 95% confidence interval: 12.2-17.0) vs. on-line eNO 13.8 ppb (95% confidence interval: 11.6-16.5). On-line and off-line results were highly correlated (r = 0.996, P = 0.000) and had similar coefficients of variation (on-line eNO 2.6%, off-line 2.8%). Neither agreement nor repeatability of eNO measurements were affected by disease status or baseline FEV(1) (% predicted values). Once standardized, the off-line eNO technique using 1-L gas collection bags will provide results similar to those recorded on-line. PMID- 11477734 TI - Efficacy of early piston-type high-frequency oscillatory ventilation in infants with respiratory distress syndrome. AB - We investigated whether the combination of surfactant replacement therapy and early application of high-frequency oscillatory ventilation (HFOV) was more effective in patients with respiratory distress syndrome (RDS) than late application of HFOV and conventional mechanical ventilation (CMV). To determine this, we retrospectively reviewed the cases of 126 neonates with RDS who received surfactant replacement therapy within 4 hr after birth. Patients were grouped into those who received HFOV immediately after birth (HFOV group), those who initially were ventilated by CMV and subsequently received HFOV (CMV/HFOV group), and those who did not receive HFOV (CMV group). Changes in respiratory system compliance (Crs), arterial-alveolar oxygen gradient (a/ApO(2)), and mean airway pressure (MAP) were compared. Infants who received HFOV were less mature than those who received CMV. The a/ApO(2) measured immediately after birth before surfactant replacement therapy was significantly lower in the HFOV and CMV/HFOV group than in the CMV group. After 72 hr, the Crs in the HFOV group was higher than in any other group and was significantly higher than the CMV/HFOV group at 48 and 120 hr. These results suggest that initiating HFOV in combination with surfactant replacement therapy immediately after birth provides effective ventilatory support for infants with RDS. PMID- 11477735 TI - Bronchiectasis: still a problem. AB - The prevalence of bronchiectasis (BR) has decreased significantly in industrialized countries, but is still commonplace in developing countries. We evaluated the causes and clinical features of BR in 23 children (13 boys (57%) and 10 girls (43%), with a mean age of 8.45 +/- 4.02 years). Infection was the major cause of BR in our region. In 8 patients, BR developed after tuberculosis or pneumonia, was associated with immune deficiency syndromes in 4 children, and with asthma in 4. Cystic fibrosis was diagnosed in 4 cases and ciliary dyskinesia in 3. In 10 patients, only one lobe was involved. Bronchiectatic lesions were most commonly found in the left lower lobe and were observed in 7 patients. Multilobar involvement was found in 13 patients. The initial treatment was primarily medical, but in 2 patients whose medical therapy failed, pulmonary resection was carried out. Three patients died from severe pulmonary infection and respiratory failure. PMID- 11477736 TI - Pulmonary involvement in a child with ligneous conjunctivitis and homozygous type I plasminogen deficiency. AB - Ligneous conjunctivitis (LC) is a rare disorder characterized by a chronic course of recurrent membranous lesions at conjunctivae. Pseudomembranes of other mucous membranes have been reported in patients with LC, but to the best of our knowledge, no case with alveolar involvement has been described. Here, we report a 2.5-year-old girl with LC who had tracheoaveolar involvement and homozygous type I plasminogen deficiency. Tracheal involvement was diagnosed by bronchoscopic biopsy and alveolar involvement with postmortem biopsy. She was shown to be homozygous for frameshift mutation in plasminogen exon 14 (Gly565ins G) with molecular genetic examination of DNA which was obtained from parafin embedded postmortem lung tissue. Ligneous inflammation of the upper and lower respiratory tract must be considered in children with LC and recurrent respiratory tract problems. PMID- 11477737 TI - Congenital pulmonary lymphangiectasia: a case report of thoracic duct agenesis. AB - We present a 17-year-old Caucasian male with congenital pulmonary lymphangiectasia and an absent thoracic duct. This patient is unique as he did not present with the disorder until age 9.5 years. Since his initial presentation he has had recurrent chylothoraces and has been treated symptomatically. We discuss the possible implications of his disorder as well as some of the limited treatment that is available. PMID- 11477738 TI - Lessons learned during 15 years of clinical information system experience. AB - This article describes lessons learned during an initial intensive care unit point-of-care clinical information system implementation and subsequent expansions to other units and hospitals in a multihospital healthcare delivery system. Although the implementation and expansions were primarily successful, lessons learned include developing a broad base of support, making decisions through consensus, addressing conflict when it occurs, keeping user expectations realistic, preparing for the change process, implementing the computer information system in stages, challenging existing work processes, viewing the implementation as a process, and choosing a project leader with outstanding communication and group process skills in addition to technical skills. PMID- 11477739 TI - Creating a tailored, multimedia, computer-based intervention. AB - Tailoring of health-related messages is effective in motivating behavior change. However, planning, designing, and implementing a tailored health promotion program can be a challenge. This article describes the process of creating a tailored multimedia program for presentation via computer to promote factory workers' use of hearing-protection devices to prevent noise-induced hearing loss. The creation process was segmented into 5 phases: 1) determination of program content; 2) creation of overall program design, including live action and still video and addition of sound and graphics; 3) development and integration of tailoring pathways into the final intervention; 4) development of a database and participant handouts; and 5) pilot testing of the intervention program. The approach used in this research program can serve as a prototype for future development of computerized multimedia health promotion interventions and can assist others in the effective use of this medium. PMID- 11477740 TI - Interface design and cognitive style in learning an instructional computer simulation. AB - An experimental design was used to investigate how cognitive style interacts with interface design to affect users' abilities to learn to use a computer simulation. Eighteen nursing students were assigned to two groups, based on their cognitive style, and asked to solve 3 physiologic problems using 3 interface designs: a strip-chart display, an integrated balloon display, and an etiologic display. Students were given up to 2 minutes to solve each problem by administering 6 different hypothetical drugs targeted at different aspects of the simulated hemodynamic system. A mixed-design analysis of variance was used to determine the effects of interface design and cognitive style on number of problems solved, time to initiate treatment, percentage of time system maintained within normal parameters, and number of drugs used. We found that the effects of cognitive style on performance were mediated by interface design and tended to decrease with practice. PMID- 11477741 TI - A third study on predicting NCLEX success with the HESI Exit Exam. AB - This was the third annual validity study designed to assess the accuracy of the HESI Exam (E2) in predicting NCLEX success for graduating registered and practical nursing students. As in year I (N = 2,725) and year II (N = 3,752), in year III (N = 6,277), the E2 was highly predictive of NCLEX success for associate degree nursing, bachelor of science nursing, diploma, and practical nursing students. Unlike previous years, in year III, monitoring was not a significant factor in the predictive accuracy of the E2. NCLEX success of low-scoring E2 students, first examined in year II, was also examined in year III. As in year II, low-scoring E2 students were significantly more (P = .001) likely to fail the licensure examination than high-scoring E2 students. In year III, unlike year II, there was no significant difference in the pass rate of low-scoring E2 students who participated in a remediation program and those who did not. The authors recommended that a more definitive definition of remediation be used in future studies and that such studies focus on E2 implementation strategies and their relationship to NCLEX success. PMID- 11477742 TI - Unmet clinical needs in dialysis: what can we do? PMID- 11477743 TI - New technologies for patients with end-stage renal disease: the US regulatory perspective. PMID- 11477744 TI - New developments in hemodialyzers. PMID- 11477745 TI - Computerized selection of membranes and hemodialysers. PMID- 11477746 TI - History of sorbents in uremia. PMID- 11477747 TI - Adsorption techniques and the use of sorbents. PMID- 11477748 TI - Characterization of flow-dynamic pattern in a new sorbent cartridge for combined hemoperfusion-hemodialysis. PMID- 11477749 TI - First clinical experience with an adjunctive hemoperfusion device designed specifically to remove beta 2-microglobulin in hemodialysis. PMID- 11477750 TI - Rationale for combined hemoperfusion/hemodialysis in uremia. PMID- 11477751 TI - Use of sorbents in acute renal failure and sepsis. PMID- 11477752 TI - The role of middle molecules in uremia--preliminary report of an international survey of nephrologists and scientists. PMID- 11477753 TI - Are new toxins appearing on the horizon? PMID- 11477754 TI - Redesigning the map of uremic toxins. PMID- 11477755 TI - Reactive carbonyl compounds as uremic toxins. PMID- 11477756 TI - Haemodialysis membranes: a matter of fact or taste? PMID- 11477757 TI - A simplified approach for real-time detection of arterial wall velocity and distension. AB - Arterial stiffness is known to increase with age and with many vascular diseases, but its noninvasive assessment in patients still represents a difficult task. The measurement of diameter change during the cardiac cycle (distension) has been proposed as a means to estimate arterial compliance and stiffness. Therefore, we have developed a simple PC-based device and algorithm for noninvasive quantification of vessel wall motion and diameter change in humans. This goal is achieved in real-time by processing the base-band signals from a commercial ultrasound Doppler system. Real-time operation is of crucial importance, because it allows a rapid achievement of optimal measurement conditions. The system was evaluated in a laboratory using a string phantom and was tested on the carotid arteries of 10 volunteers. Wall velocities from 0.05 to 600 mm/s and displacements lower than 2 microns were detected with phantoms. The measured carotid diameter change in the volunteers ranged from 7.5 to 11.8% (mean = 9.8%) and agrees closely with values reported in the literature. The difference between values taken one hour apart ranged from 0.2 to 0.5%. We conclude that the new system provides rapid, accurate, and repeatable measurements of vessel distension in humans. PMID- 11477758 TI - Three-stage approach to ultrasound contrast detection. AB - A new method for detecting ultrasound contrast agents using a three-stage pulsing sequence is proposed. The method is based on observations showing that the scattering properties of contrast agents are modified by ultrasonic insonation at high power, but remain unchanged at low power. The objective of the first stage of the pulsing sequence is to use low power pulses to obtain a high resolution reference image without altering the agent. Higher power pulses in the second stage modify the contrast agent. The third stage detects the changes imposed to the contrast agent using low power pulses. A temporal filter is proposed to discriminate contrast response from clutter signal. The method is similar to power Doppler methods in that it uses several pulses to survey the target while destroying the agent. The new idea is to separate detection and destruction to circumvent a trade-off between sensitivity and resolution. Results from in vitro experiments with three different contrast agents are presented. The results are compared with harmonic power Doppler processed from the same data and show that an improvement in sensitivity is achievable by including the high power burst in the pulsing sequence. The results also show that the proposed filter reduces clutter artifacts from moving tissue. PMID- 11477759 TI - Ultrasound scattering model: 2-D cross-correlation and focusing criteria--theory, simulations, and experiments. AB - A microscopic scattering model is developed to expedite simulation studies of ultrasound imaging in soft tissue using multichannel transducer probes. The model fully accounts for the physics of broadband signals, propagating wave packets, and time delay focusing. Analytical results are presented for 2-D transducer arrays; 1-D results can be trivially extracted by setting the number of rows equal to unity. The 2-D cross-correlation and the 2-D form of the Mallart-Fink (MF) focusing factor are calculated. It is demonstrated that the scattering model reduces to the 2-D form of the monochromatic van Cittert Zernike (VCZ) analysis. Simulation results for the focusing factor are presented, and comparisons are given between the values obtained from simulation, analytical theory, and actual water tank experiments. The comparative results are all in close accord with each other. PMID- 11477760 TI - Ultrasonic six-axis deformation sensing. AB - In this paper, we describe a newly developed deformation sensing scheme in a soft medium, which is based on precise encoding and decoding of deformation components into ultrasound wavefronts. It can detect three translational components and three rotational components of displacement around a transmitter position nearly simultaneously. We assume a cell structure that consists of a 2 x 2 ultrasonic transmitter matrix and a 2 x 2 ultrasonic receiver matrix, which are placed face to face at a distance of a few tens of wavelengths. All of the transmitter elements are driven sinusoidally and simultaneously, but they are switched into the same, reversed, or quadrature phases to generate a particular shape of wavefront on the receiver matrix. The receiver elements are connected in such a way to obtain amplitude and spatial gradients of the wavefront at a center of the receiver matrix. First, we describe the transduction theory for the six dimensions and show the orthogonality, locality, and simultaneity of this sensing scheme. Then, we describe the fabrication and experimental evaluation of the cell. We also describe a prototype tactile sensor in which a single cell is embedded in a flexible hemispherical fingertip-like body. PMID- 11477761 TI - The generation and detection of longitudinal guided waves in thin fibers using a conical transformer. AB - This paper describes a technique to couple ultrasonic energy from a piezoceramic disc transducer into a fiber waveguide to induce longitudinal propagation. A polymer cone is utilized to bond the fiber waveguide onto the surface of the disc and to behave as a mechanical transformer, converting lateral displacements at its base into longitudinal displacements at its apex. Wideband finite element analysis (FEA) results are provided to show that the bond efficiently couples the radial modes of a disc transducer into fiber waveguides for longitudinal mode excitation. Furthermore, narrowband FEA is utilized to investigate how the geometry and material properties of the bond and waveguide influence the coupling efficiency. The technique is then quantified in terms of signal-to-coherent noise ratio (SCNR), reflecting its ability to generate the desired longitudinal waveguide mode and reject erroneous modes. Finally, design parameters are outlined for the successful implementation of this technique. PMID- 11477762 TI - Traveling wave excitation in a flexural vibration ring by using a torsional flexural composite transducer. AB - This paper presents a new mechanism for traveling wave excitation of the flexural vibration in a ring. A torsional-flexural composite transducer was used to excite two degenerate flexural vibration modes spatially and temporally orthogonal to each other in the ring. The proto-type, composed of a 32-mm diameter composite transducer attached along the outer edge of an 180-mm diameter aluminum annular ring, was designed, constructed, and tested. The ring was designed via finite element analysis to operate using a (0.8) transverse flexural traveling wave, and the motion was confirmed experimentally. PMID- 11477763 TI - Improved ultrasonic spectroscopy methods for characterization of dispersive materials. AB - Transmission ultrasonic spectroscopy method has been successfully implemented for the characterization of piezoceramics at high frequencies. There are, however, still some intrinsic error sources that limit the accuracy of the method. In this paper, two improved ultrasonic spectroscopy methods are presented, which can reduce the number of pre-required parameters and reduce another error source. The two improved methods were used to measure the frequency dispersion of phase velocity and attenuation of doped piezoceramic lead zirconate titanate (PZT-5A); results were compared with those obtained from the conventional method. The advantages and limitations of each method are discussed. PMID- 11477764 TI - Longitudinal-bending mode micromotor using multilayer piezoelectric actuator. AB - Longitudinal-bending mode ultrasonic motors with a diameter of 3 mm were fabricated using stacked multilayer piezoelectric actuators, which were self developed from hard lead zirconate titanate (PZT) ceramic. A bending vibration was converted from a longitudinal vibration with a longitudinal-bending coupler. The motors could be bidirectionally operated by changing driving frequency. Their starting and braking torque were analyzed based on the transient velocity response. With a load of moment of inertia 2.5 x 10(-7) kgm2, the motor showed a maximum starting torque of 127.5 microNm. The braking torque proved to be a constant independent on the motor's driving conditions and was roughly equivalent to the maximum starting torque achievable with our micromotors. PMID- 11477765 TI - A programmable ultra-low noise X-band exciter. AB - A programmable ultra-low noise X-band exciter has been developed using commercial off-the-shelf components. Its phase noise is more than 10 dB below the best available microwave synthesizers. It covers a 7% frequency band with 0.1-Hz resolution. The X-band output at +23 dBm is a combination of signals from an X band sapphire-loaded cavity oscillator (SLCO), a low noise UHF frequency synthesizer, and special-purpose frequency translation and up-conversion circuitry. PMID- 11477766 TI - A stratified model to predict dispersion in trabecular bone. AB - Frequency-dependent phase velocity (dispersion) has previously been measured in trabecular bone by several groups. In contrast to most biologic tissues, phase velocity in trabecular bone tends to decrease with frequency. A stratified model, consisting of alternating layers of bone and marrow (in vivo) or water (in vitro), has been employed in an attempt to explain this phenomenon. Frequency dependent phase velocity was measured from 300 to 700 kHz in 1) phantoms consisting of regularly spaced thin parallel layers of polystyrene sheets in water and 2) 30 calcaneus samples in vitro. For the polystyrene phantoms, the agreement between theory and experiment was good. For the calcaneus samples, the model has some limited usefulness (uncertainty of about 5%) in predicting average phase velocity. More importantly, the model seems to perform consistently well for predicting the frequency dependence of phase velocity in calcaneus. PMID- 11477767 TI - On velocity estimation using speckle decorrelation. AB - Quantitative estimation of blood velocity using Doppler techniques is fundamentally limited because only the axial component can be detected. Speckle decorrelation resulting from scatterer motion may be used to compute non-axial components and to obtain quantitative flow information. Based on both simulations and experimental results, it is shown that the decorrelation technique is feasible only for constant flows. If flow gradients are present, the correlation between two signals along the same line of observation may be significantly affected by the gradients. Therefore, the decorrelation method cannot be used for quantitative flow estimation if flow gradients are not accurately measured and effects on signal correlation are not fully compensated. Results in this paper show that accurate estimation of flow gradients is practically difficult. It is further shown that effects of signal-to-noise ratio (SNR) on the correlation must also be taken into account for quantitative flow analysis. PMID- 11477768 TI - Parametric study of the peak negative acoustic pressure distribution within the image plane of a phased array transducer. AB - A study is presented in which the influence of pulse and transducer characteristics on the distribution of the peak negative acoustic pressure within the image plane of a phased array transducer is shown. First, the influence of the center frequency, the bandwidth of the transmitted pulse, the position of the focus point, and the properties of the phased array are investigated by computer simulation. These simulations show that both the maximal amplitude and the homogeneity of the distribution of the peak negative pressure within a sector image depend on these parameters. Most remarkably, they show that the maximal peak pressure does not necessarily occur on the geometrical symmetry axis of the array transducer but can occur at the outermost lateral lines of a sector image. Second, this effect is demonstrated experimentally by hydrophone measurements of the sound field produced by a standard, clinically used, 2.5-MHz phased array transducer. The experimental results agree well with the theoretical model. The relevance of this study for contrast echography and the safety measurements of ultrasound is discussed. PMID- 11477769 TI - Deflection characteristics of a trapezoidal multilayer in-plane bending piezoelectric actuator. AB - A novel multilayer split-morph actuator has been designed and fabricated using the thick film screen-printing technology. Deflection characteristics of the split-morph actuator have been investigated by theoretical analysis and experimental measurement. The results indicate that the tip displacement is inversely proportional to the thickness of each piezoelectric layer, but is independent of the number of layers and the total thickness of the actuator. The displacement/voltage sensitivity of the trapezoidal actuator is larger than that of the rectangular design, assuming both have the same width of clamped end and the same thickness of the piezoelectric layers. The maximum displacement/voltage sensitivity of 0.157 micron/V was obtained with a split-morph actuator with 30 micron thick layers. The proposed actuator is a promising candidate for the secondary fine-tuning actuator of a dual stage head-positioning servo system in high density hard disk drives. PMID- 11477770 TI - Strain rate imaging using two-dimensional speckle tracking. AB - Strain rate images (SRI) of the beating heart have been proposed to identify non contracting regions of myocardium. Initial attempts used spatial derivatives of tissue velocity (Doppler) signals. Here, an alternate method is proposed based on two-dimensional phase-sensitive speckle tracking applied to very high frame rate, real-time images. This processing can produce high resolution maps of the time derivative of the strain magnitude (i.e., square root of the strain intensity). Such images complement traditional tissue velocity images (TVI), providing a more complete description of cardiac mechanics. To test the proposed approach, SRI were both simulated and measured on a thick-walled, cylindrical, tissue equivalent phantom modeling cardiac deformations. Real-time ultrasound images were captured during periodic phantom deformation, where the period was matched to the data capture rate of a commercial scanner mimicking high frame rate imaging of the heart. Simulation results show that SRI with spatial resolution between 1 and 2 mm are possible with an array system operating at 5 MHz. Moreover, these images are virtually free of angle-dependent artifacts present in TVI and simple strain rate maps derived from these images. Measured results clearly show that phantom regions of low deformation, which are difficult to identify on tissue velocity-derived SRI, are readily apparent with SRI generated from two-dimensional phase-sensitive speckle tracking. PMID- 11477771 TI - Modeling of ultrasonic wave propagation in teeth using PSpice: a comparison with finite element models. AB - Ultrasound is used extensively in the medical field for the detection and characterization of a variety of features in the human body. Finite element models used to understand ultrasonic wave propagation in teeth have been developed so that ultrasound techniques could be realized in dentistry. This paper presents a hypothesis that underlies one possible design of an ultrasonic tool that can be used in a clinical environment, as well as several models that describe acoustic field simulation, propagation, and interaction with the layers of several tooth structures. A complete PSpice model of a single-element transducer based on Redwood's version of Mason's equivalent circuit, a focusing lens, and a multi-layer tooth structure is used to illustrate the validity of this hypothesis. Transmission line theory is employed as a basis for the models of the piezoceramic, the lens, and the different tooth layers. Results clearly depict the transmission and reflection of the ultrasonic waves as they travel through the layers within the tooth structure and point out the noticeable similarity to longitudinal L-wave signatures produced by axisymmetric finite element models presented in earlier studies. PMID- 11477772 TI - Investigation of crossed SAW fields by scanning acoustic force microscopy. AB - We used multimode scanning acoustic force microscopy (SAFM) for studying noncollinearly propagating Rayleigh and Love wave fields. By analyzing torsion and bending movement of SAFM cantilever, normal and in-plane wave oscillation components are accessible. The SAFM principle is the down-conversion of surface oscillations into cantilever vibrations caused by the nonlinearity of the tip sample interaction. Through mixing of complementary oscillation components, phase velocities of crossed Rayleigh waves on GaAs and crossed Rayleigh and Love waves on the layered system SiO2/ST-cut quartz were obtained simultaneously. Now, it is possible to investigate elastic properties of submicron areas through multimode SAFM measurements. Finally, we present mixing experiments of four SAWs on GaAs and discuss the various influences on the measured SAFM amplitude and phase contrast. PMID- 11477773 TI - Use of frequency diversity and Nakagami statistics in ultrasonic tissue characterization. AB - The Nakagami distribution was recently proposed as a generalized model for the envelope of the backscattered ultrasonic echo from tissue. The parameters of the Nakagami model were also shown to be useful in tissue characterization. This paper explores the possibility of enhancing the ability of these parameters for tissue characterization through the techniques of diversity and compounding. Frequency diversity has been used to create multiple versions of the envelope, which are then combined. This compounded envelope has been modeled, and its parameters have been analyzed. The ability of these new parameters to enhance tissue characterization is studied using computer simulation and experiments on tissue-mimicking phantoms. Results indicate that the use of frequency diversity and compounding may indeed improve the ability of the parameters of the Nakagami model to separate different number densities of scatterers. Therefore, it is suggested that such an approach may lead to better techniques in ultrasonic tissue characterization. PMID- 11477775 TI - Comments on a study of drift in cesium frequency standards. AB - This letter comments on a recent study of the drift rate of 115 cesium standards. The conclusion here is that no aging effect is visible in the results of the study. PMID- 11477774 TI - Vibration characteristics of composite piezoceramic plates at resonant frequencies: experiments and numerical calculations. AB - The experimental measurement of the resonant frequencies for the piezoceramic material is generally performed by impedance analysis. In this paper, we employ an optical interferometry method called the amplitude-fluctuation electronic speckle pattern interferometry (AF-ESPI) to investigate the vibration characteristics of piezoceramic/aluminum laminated plates. The AF-ESPI is a powerful tool for the full-field, noncontact, and real-time measurement method of surface displacement for vibrating bodies. As compared with the conventional film recording and optical reconstruction procedures used for holographic interferometry, the interferometric fringes of AF-ESPI are produced instantly by a video recording system. Because the clear fringe patterns measured by the AF ESPI method will be shown only at resonant frequencies, both the resonant frequencies and corresponding vibration mode shapes are obtained experimentally at the same time. Excellent quality of the interferometric fringe patterns for both the in-plane and out-of-plane vibration mode shapes are demonstrated. Two different configurations of piezoceramic/aluminum laminated plates, which exhibit different vibration characteristics because of the polarization direction, are investigated in detail. From experimental results, we find that some of the out of-plane vibration modes (Type A) with lower resonant frequencies cannot be measured by the impedance analysis; however, all of the vibration modes of piezoceramic/aluminum laminated plates can be obtained by the AF-ESPI method. Finally, the numerical finite element calculations are also performed, and the results are compared with the experimental measurements. Excellent agreements of the resonant frequencies and mode shapes are obtained for both results. PMID- 11477776 TI - Two-dimensional noise-robust blind deconvolution of ultrasound images. AB - This paper presents a new method for 2-D blind homomorphic deconvolution of medical B-scan ultrasound images. The method is based on noise-robust 2-D phase unwrapping and a noise-robust procedure to estimate the pulse in the complex cepstrum domain. Ordinary Wiener filtering is used in the subsequent deconvolution. The resulting images became much sharper with better defined tissue structures compared with the ordinary images. The deconvolved images had a resolution gain along the order of 3 to 7, and the signal-to-noise ratio (SNR) doubled for many of the images used in our experiments. The method gave stable results with respect to noise and grey levels through several image sequences. PMID- 11477777 TI - Three-dimensional blind deconvolution of ultrasound images. AB - Three-dimensional ultrasound images are blurred by the ultrasound pulse through the convolution between the 3-D tissue signal and the 3-D pulse. The blurring reduces the spatial resolution of the 3-D ultrasound images and, consequently, their diagnostic value. This paper presents a method for 3-D blind homomorphic deconvolution of medical 3-D ultrasound images to improve their spatial resolution. The blind estimate of the 3-D pulse is necessary because the pulse changes in spatial extent and frequency composition as it passes through the tissues and because the pulse is not separable in its spatial dimensions. The method was tested on a 3-D image of a phantom with anechoic spheres of known size in a uniform diffuse scattering matrix. The spheres were clearly better defined and had volumes much closer to the true volume in the deconvolved image than in the original image. PMID- 11477778 TI - Finite element simulation of piezoelectric transformers. AB - Piezoelectric transformers are nothing but ultrasonic resonators with two pairs of electrodes provided on the surface of a piezoelectric substrate in which electrical energy is carried in the mechanical form. The input and output electrodes are arranged to provide the impedance transformation, which results in the voltage transformation. As they are operated at a resonance, the electrical equivalent circuit approach has traditionally been developed in a rather empirical way and has been used for analysis and design. The present paper deals with the analysis of the piezoelectric transformers based on the three dimensional finite element modelling. The PIEZO3D code that we have developed is modified to include the external loading conditions. The finite element approach is now available for a wide variety of the electrical boundary conditions. The equivalent circuit of lumped parameters can also be derived from the finite element method (FEM) solution if required. The simulation of the present transformers is made for the low intensity operation and compared with the experimental results. Demonstration is made for basic Rosen-type transformers in which the longitudinal mode of a plate plays an important role; in which the equivalent circuit of lumped constants has been used. However, there are many modes of vibration associated with the plate, the effect of which cannot always be ignored. In the experiment, the double resonances are sometimes observed in the vicinity of the operating frequency. The simulation demonstrates that this is due to the coupling of the longitudinal mode with the flexural mode. Thus, the simulation provides an invaluable guideline to the transformer design. PMID- 11477779 TI - Optical resonant ultrasound spectroscopy for fluid properties measurement. AB - The properties of fluids are studied using unusually small containment spherical resonators. Proper identification of resonant fluid signatures allows determination of pressure and density of the internal gas with great accuracy using an appropriate equation of state (EOS). Low noise and high sensitivity detection of vibration are critical parameters to characterizing the contained gas when its pressure approaches 1 atm or less. The benefits of using spherical resonators to determine fluid properties are discussed, and some example calculations of sound speed are presented. In addition to measuring fluids, a comparative experimental approach is taken to explore and, eventually, to optimize vibration detection. In the experiments, two detection methods, a contact piezoelectric transducer (PZT) device and a non-contact optical device, are compared simultaneously and quantitatively. This is done in a unique manner without change in vibration coupling to the sample between tests. A commercially available resonant ultrasound spectroscopy system is used as the contact system, while another commercial device (used as the non-contact vibration detector) combined with the same excitation source (used in the contact system) comprises the other system. The non-contact detector is an optical interferometric receiver that provides adaptation to optically rough surfaces and high sensitivity to acoustic displacements through optical interference in photorefractive GaAs. Both vibration detection systems are compared with particular emphasis on displacement sensitivity, frequency response, and noise level. Furthermore, the results from comparing detection modalities are presented, and their effects on fluid properties measurement are discussed. PMID- 11477780 TI - A new estimator for vector velocity estimation. AB - A new estimator for determining the two-dimensional velocity vector using a pulsed ultrasound field is derived. The estimator uses a transversely modulated ultrasound field for probing the moving medium under investigation. A modified autocorrelation approach is used in the velocity estimation. The new estimator automatically compensates for the axial velocity when determining the transverse velocity. The estimation is optimized by using a lag different from one in the estimation process, and noise artifacts are reduced by averaging RF samples. Further, compensation for the axial velocity can be introduced, and the velocity estimation is done at a fixed depth in tissue to reduce the influence of a spatial velocity spread. Examples for different velocity vectors and field conditions are shown using both simple and more complex field simulations. A relative accuracy of 10.1% is obtained for the transverse velocity estimates for a parabolic velocity profile for flow transverse to the ultrasound beam and a SNR of 20 dB using 20 pulse-echo lines. The overall bias in the estimates was -4.3%. PMID- 11477781 TI - Finite element analysis of the behavior of the Scanning Microdeformation Microscope. AB - The Scanning Microdeformation Microscope, as many other scanning probe microscopes developed in the last years, is a kind of ac force microscope using the near-field acoustic interaction. The heart of the system is an electromechanical oscillator made of a silicon cantilever, a diamond or sapphire tip, associated with a bimorph piezoelectric transducer and a specific amplifier. The specificity of the system is the way of detection of the oscillation frequency performed electrically through the admittance of the piezoelectric transducer. In this paper, we describe the technique of detection involved in the microscope. A modelling of the complete behavior of the electromechanical oscillator performed with the finite element method (FEM) (of simulation) is presented. A comparison between experimental and theoretical behavior shows a very good agreement. PMID- 11477782 TI - Recurrent fuzzy neural network control for piezoelectric ceramic linear ultrasonic motor drive. AB - In this study, a recurrent fuzzy neural network (RFNN) controller is proposed to control a piezoelectric ceramic linear ultrasonic motor (LUSM) drive system to track periodic reference trajectories with robust control performance. First, the structure and operating principle of the LUSM are described in detail. Second, because the dynamic characteristics of the LUSM are nonlinear and the precise dynamic model is difficult to obtain, a RFNN is proposed to control the position of the moving table of the LUSM to achieve high precision position control with robustness. The back propagation algorithm is used to train the RFNN on-line. Moreover, to guarantee the convergence of tracking error for periodic commands tracking, analytical methods based on a discrete-type Lyapunov function are proposed to determine the varied learning rates of the RFNN. Then, the RFNN is implemented in a PC-based computer control system, and the LUSM is driven by a unipolar switching full bridge voltage source inverter using LC resonant technique. Finally, the effectiveness of the RFNN-controlled LUSM drive system is demonstrated by some experimental results. Accurate tracking response and superior dynamic performance can be obtained because of the powerful on-line learning capability of the RFNN controller. Furthermore, the RFNN control system is robust with regard to parameter variations and external disturbances. PMID- 11477783 TI - Frequency-dependent open-circuit acoustic sensitivity of fluid-filled, coated, radially polarized piezoelectric ceramic cylindrical shells of arbitrary thickness and infinite length. AB - The frequency-dependent free-field open-circuit voltage generated by radially polarized piezoelectric ceramic fluid-filled shells of infinite length coated with an elastic layer and excited by a plane acoustic wave is studied. The shell and the coating are of arbitrary thickness and are anisotropic. It is shown that the charge density in the piezoelectric shell is zero by using the electrostatic and open-circuit conditions. General expressions for the pressures and radial velocities in the interior and exterior fluids are obtained by using the finiteness and radiation conditions, respectively. General expressions for the radial stresses and velocities in the piezoelectric shell and elastic coating are also determined by using two-dimensional equations of state and axisymmetric equations of motion. The coefficients in the general expressions are then determined by using the continuity conditions at the various interfaces. Finally, an expression for the open-circuit sensitivity is obtained by using a piezoelectric equation of state. Numerical results are presented for air, water, and vacuum in the interior, two types of elastic coating, and piezoelectric shells with various thicknesses. The effect of neglecting anisotropy on the sensitivity is also illustrated. PMID- 11477784 TI - Digital phased array beamforming using single-bit delta-sigma conversion with non uniform oversampling. AB - Digital beamforming based on oversampled delta-sigma (delta sigma) analog-to digital (A/D) conversion can reduce the overall cost, size, and power consumption of phased array front-end processing. The signal resampling involved in dynamic delta sigma beamforming, however, disrupts synchronization between the modulators and demodulator, causing significant degradation in the signal-to-noise ratio. As a solution to this, we have explored a new digital beamforming approach based on non-uniform oversampling delta sigma A/D conversion. Using this approach, the echo signals received by the transducer array are sampled at time instants determined by the beamforming timing and then digitized by single-bit delta sigma A/D conversion prior to the coherent beam summation. The timing information involves a non-uniform sampling scheme employing different clocks at each array channel. The delta sigma coded beamsums obtained by adding the delayed 1-bit coded RF echo signals are then processed through a decimation filter to produce final beamforming outputs. The performance and validity of the proposed beamforming approach are assessed by means of emulations using experimental raw RF data. PMID- 11477785 TI - A new detection method for capacitive micromachined ultrasonic transducers. AB - Capacitive micromachined ultrasonic transducers (cMUT) have become an alternative to piezoelectric transducers in the past few years. They consist of many small circular membranes that are connected in parallel. In this work, we report a new detection method for cMUTs. We model the membranes as capacitors and the interconnections between the membranes as inductors. This kind of LC network is called an artificial transmission line. The vibrations of the membranes modulate the electrical length of the transmission line, which is proportional to the frequency of the signal through it. By measuring the electrical length of the artificial line at a high RF frequency (in the gigahertz range), the vibrations of the membranes can be detected in a very sensitive manner. For the devices we measured, we calculated the minimum detectable displacement to be in the order of 10(-5) A/square root of Hz with a possible improvement to 10(-7) A/square root of Hz. PMID- 11477786 TI - Optimization of wide-band linear arrays. AB - An optimization method is proposed for linear arrays to be used in ultrasound systems under wide-band operation. A fast algorithm, the threshold accepting, has been utilized to determine the element positions and weight coefficients of a linear array that generates a desired beam pattern. To reduce the computational burden in the optimization procedure, an efficient numerical routine for the beam pattern evaluation has been implemented. We address the optimization problem of both dense and sparse wide-band arrays. In the first case, the goal is to minimize the side-lobe energy by varying the element weights; we compare the optimized beam pattern with that obtained with classical shading functions, showing that better results can be achieved with a wide-band optimization. We also consider the optimization of the layout (positions and weights) of a sparse linear array to achieve a desired beam pattern with a fixed or minimum number of array elements. The comparison of the proposed method with a narrow-band optimization algorithm is presented, showing that better performances (about -7 dB further reduction of the side-lobe level) can be achieved with a wide-band sparse array optimization. Further numerical simulations are given, showing that the proposed method yields better results than wide-band sparse random arrays and periodic arrays with the same aperture width. PMID- 11477787 TI - On modeling biomedical ultrasound RF echoes using a power-law shot-noise model. AB - We propose a new model for the RF ultrasound echo, namely the power-law shot noise process. Based on this model, the in-phase and quadrature components of the echo are shown to exhibit 1/f beta-type spectral behavior, in a sense that is defined in the paper. The envelope also exhibits this type of spectral behavior, but with a different exponent. This result explains the experimental observations by other researchers of the power-law trend of the RF echo spectrum. Although the shot-noise model has been used in the past for modeling the RF echo, this is the first time that a power-law impulse response filter is used and that the resulting 1/f beta-type spectral behavior of the RF echo has been investigated. The model parameters are linked to tissue characteristics, such as scatterer density and attenuation; thus, they have the potential to be used as tissue characterization features. The validity of the proposed model is tested based on a database of 100 clinical ultrasound images of the breast. PMID- 11477788 TI - Acoustical diffraction tomography in a finite form and its computer simulations. AB - Until now, all acoustical diffraction reconstruction algorithms are in infinite forms. All of these algorithms have disadvantages: severe limitations on scatterers or tedious calculations. In this paper, we present a new reconstruction algorithm in a finite form using the method of formal parameter, which is very simple. This new algorithm gives an exact reconstruction when the amplitude of the scattered wave is smaller than that of the incident wave everywhere. This assumption is much less restrictive than that for the first- and second-order Born approximations. Although this new algorithm is in a finite form, it is still an approximate one when the amplitude of the scattered wave is not smaller than that of the incident wave everywhere. However, it still gives a good reconstruction when the amplitude of the scattered wave is a little greater than that of the incident wave in some area. Some numerical examples have confirmed these conclusions. PMID- 11477789 TI - Admittance matrix of asymmetric piezoelectric bimorph with two separate electrical ports under general distributed loads. AB - The dynamic admittance matrix of the asymmetric triple-layer piezoelectric bimorph subjected to the general distributed harmonic loads as well as the flexural moments and the vertical loads at the tip are presented. The top and bottom piezoelectric layers have two separate electrical ports such that each layer can be used as either a sensor or an actuator. The variation principle is used for deriving the motion equations and the conjugate parameters that maintain the symmetry of the admittance matrix. The mechanical displacements and forces at the tip are expressed in a matrix form, which, together with the reciprocal condition, greatly simplify the analysis procedure. The derived admittance matrix under the cantilevered condition is presented by a five-by-five matrix, each row representing the relationships of the displacement and rotation at the tip, the volume averaged displacement, the separate electrical charges with the flexural moment and vertical load at the tip, the magnitude of the distributed load, and the voltages. The matrices, which reduce to simpler forms for several special cases, are then used to determine the two-port electrical admittance. It is shown that the derived admittance matrix covers the various boundary conditions, the electrical parallel and series connections, and the arbitrary lay-up, including the unimorph, used as both sensors and actuators. PMID- 11477790 TI - Parametric modeling with beamspread compensation and MIMO frequency domain inversion applied to fine saturated sands. AB - A system identification technique is applied to estimate the intrinsic absorption and dispersion of two fine sands. The method is based on the parametric modeling of the wave propagation through a Plexiglas tank filled with the sediment under investigation. The applicability of various porous models is discussed. The viscoelastic constant Q model and viscoelastic rational form model are applied and compared. Closed form expressions describing the wave propagation are replaced by Debye series expansions with correction coefficients that consider the beamspread caused by the finite aperture of the emitter. A multiple input multiple output (MIMO) representation is used in conjunction with the maximum likelihood estimator (MLE) for the estimation of the sediment parameters. The estimated absorption and dispersion curves are depicted. PMID- 11477791 TI - Surface acoustic waves propagating over a rotating piezoelectric half-space. AB - Surface acoustic waves (SAW) propagating over a piezoelectric half-space rotating at a constant angular rate about a fixed axis are analyzed using the linear theory of piezoelectricity, including Coriolis and centrifugal forces. Rotation sensitivity, the rotation induced change of wave speed, is studied. The dependence of the rotation sensitivity on the orientation of the rotation axis and the orientation of the material is examined. Numerical results for polarized ceramics PZT-5H are presented to show the detailed characteristics of the rotation sensitivity. The implications of the numerical results are discussed for different applications. PMID- 11477792 TI - [Introducing the 21st century by modernizing the obstetrics and gynecology--the 50 years' achievement and prospect of the obstetrics and gynecology]. PMID- 11477793 TI - [Development of the obstetrics and gynecology in China]. PMID- 11477794 TI - [Epidemiological analysis on the risk factors of intrauterine transmission of hepatitis virus C]. AB - OBJECTIVE: To investigate the possibility and risk factors of intrauterine transmission of hepatitis virus C (HCV) in Taiyuan City. METHODS: Anti-HCV and HCV RNA were detected by enzyme linked immunosorbent assay(ELISA) and reverse transcriptase Polymerase Chain Reaction(RT-PCR), in 64 mothers with HCV or HCV RNA positive and their newborns cord blood samples. Case-control study was used for the risk factors analysis. RESULTS: The infection rate of HCV was 66.15% in newborns from anti-HCV or HCV RNA positive mothers. The intrauterine transmission rate was 100% in newborns from HCV RNA positive mothers. There was a significant correlation between HCV intrauterine transmission and maternal history of blood transfusion and abnormal serum alanine aminotransferase(ALT), and the relative risk was 317.15 and 2.60, respectively. CONCLUSIONS: The higher incidence of intrauterine transmission was found in newborns from HCV RNA or anti-HCV positive mothers. The high risk factors of intrauterine transmission are maternal blood transfusion history and abnormal ALT. PMID- 11477795 TI - [Study on the relationship between perinatal hypoxia and concentration of endothelin-1 in amniotic fluid]. AB - OBJECTIVE: To investigate the relationship between endothelin-1 (ET-1) concentration in amniotic fluid (AF) and perinatal hypoxia. METHODS: 161 cases were measured for amniotic fluid(AF) ET-1 levels by radioimmunoassay. 110 cases of normal pregnancy were included in control group among which 30 term pregnancies were simultaneously measured for maternal and umbilical plasma ET-1. 51 cases of intrauterine hypoxia were the study group. RESULTS: (1) The AF ET-1 levels showed increasing trend after 14 weeks (P < 0.01). (2) Fetal plasma ET-1 levels were significantly higher than that of maternal plasma ET-1 levels, but lower than those of AF ET-1 (P < 0.01). The ET-1 levels of umbilical plasma had positive correlation with those of AF ET-1 (r = 0.952, P < 0.01), but there is no correlation with levels of maternal plasma ET-1 (r = 0.338, P < 0.05). (3) In study group, the level of AF ET-1 was elevated with severity of hypoxia, the average level of AF ET-1 in cases of intrauterine hypoxia was (30.654 +/- 5.832) ng/L. In cases of severe neonatal asphyxia it was (960.650 +/- 236.698) ng/L. CONCLUSIONS: The ET-1 exists in AF and gradually increases while pregnancy advanced. AF ET-1 levels can be served as a marker to predict perinatal hypoxia. PMID- 11477796 TI - [Clinical analysis of the absent and/or reverse end diastolic velocity of fetal umbilical artery]. AB - OBJECTIVE: To investigate the relations between absent and/or reverse end diastolic velocity of fetal umbilical artery (AEDV) and maternal and fetal pathological factors and perinatal outcome. METHODS: To analyze the clinical informations of 33 case, with AEDV by pulse doppler ultrasonic examination. RESULTS: (1) There are 9 cases (27.3%) complicated with different kinds of fetal morphologic abnormality, 6 cases of perinatal death. (2) In 6 cases (18.2%) of twins, twin-twin transfusion syndrome (TTTS) occurred in 3 cases. In 6 cases when the body weight differences between the 2 babies were higher than 24.8%, neonatal death occurred in 4 cases. (3) The group with AEDV showed significantly higher incidence of pregnancy induced hypertension, oligoaminos, IUGR, fetal distress and neonatal death than that of normal group. CONCLUSIONS: The appearance of AEDV shows the extreme disfunction of mothermal placental-fetal circulation and may be related to poor outcome. PMID- 11477797 TI - [Preliminary study on the relationship between human histocompatibility leukocyte antigen-G mRNA and recurrent spontaneous abortion]. AB - OBJECTIVE: To examine the expression of Human Histocompatibility Leukocyte Antigen-G (HLA-G) mRNA in placenta from cases of recurrent spontaneous abortion (RSA) and try to study the relationship between HLA-G mRNA and RSA. METHODS: In situ hybridization was performed on maternal-fetal interface sections of placenta in first trimester, third trimester of normal pregnancy and RSA women who were treated with immunotherapy and had term delivery. RESULTS: The positive signal of HLA-G expression were showed in all placentas of normal pregnant women within villus and cytoplasma of syncytiotrophoblast and cytotrophoblast. However, there is no any HLA-G mRNA signals in placentas of RSA women after immunotherapy. CONCLUSIONS: HLA-G mRNA is not the main cause of RSA and there is no direct relations to fetal survival. PMID- 11477798 TI - [Pregnancy induced hypertension complicated acute disseminated intravascular coagulation: clinical analysis of 26 cases]. AB - OBJECTIVE: To determine the relationship between pregnancy induced hypertension (PIH) and acute disseminated intravascular coagulation (DIC). METHODS: 26 cases with PIH complicated acute DIC were analyzed retrospectively in five hospitals of Xi'an from 1980 to 1997. RESULTS: (1) In 26 patients with PIH complicated acute DIC, 7 cases died (26.92%), while 17 neonates died (58.62%) in 29 neonates (3 cases were twin pregnancy). Cesarean section and hysterectomy were performed on 7 cases respectively, including 4 cases underwent both of them. (2) The causative factors of PIH complicated acute DIC included placental abruption (7 cases), amniotic fluid embolism (4 cases), eclamptic seizure (4 cases), surgical injury (7 cases) and bleeding (4 cases). 7 cases induced by placental abruption were all cured, 4 cases induced by amniotic fluid embolism all died. CONCLUSION: There are close relationship between PIH and DIC, PIH complicated acute DIC is the major cause of maternal and perinatal mortality, special attention should be paid on preventing DIC for PIH. PMID- 11477799 TI - [Dual color fluorescent in-situ hybridization detects sex chromosome mosaicism in human embryos]. AB - OBJECTIVE: Using dual color fluorescent in-situ hybridization (FISH) to detect sex chromosome mosaicism in human embryos and perform preimplantation gender diagnosis. METHODS: Three abnormal fertilized embryos and 18 normal fertilized embryos which were not suitable for embryo transfer and cryopreservation were analyzed by dual color fluorescent in-situ hybridization. RESULTS: Mosaicism analysis was performed on 14 normal fertilized embryos, which had more than two cells. Nine embryos had sex chromosome mosaicism. XO in male and female embryos cells were 12% and 14% respectively. CONCLUSION: Identification of sex by FISH analysis of a single cleavage cell is accurate, and sex chromosome mosaicism will not affect preimplantation gender diagnosis. PMID- 11477800 TI - [Placenta accreta in early pregnancy: a case report and review of the literature]. AB - OBJECTIVE: To summarize and investigate the clinical characteristics and treatment of placenta accreta in early pregnancy. METHODS: A case of placenta accreta in early pregnancy presenting severe bleeding during suction curettage was analyzed retrospectively and the literature was reviewed. RESULTS: The major risk factors for placenta accreta in early pregnancy were related to uterine scars from previous cesarean section. In most cases there was vaginal bleeding after amenorrhea. It was common that uncontrollable bleeding took place during curettage and hysterectomy was usually necessary. CONCLUSION: Cesarean section is one of the high risk factors of placenta accreta in early pregnancy. PMID- 11477801 TI - [Study on in vitro cytokines levels induced from peripheral mononuclear cells in patients with endometriosis]. AB - OBJECTIVE: To study the changes and significance of interleukin-6 (IL-6), interleukin-8(IL-8), tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) levels in vitro cultured system of peripheral mononuclear cells in patient with endometriosis(Em). METHODS: Peripheral blood mononuclear cells from 31 cases of endometriosis, classified into mild group (stage I and II, n = 11), and severe group (stage III and IV, n = 20) according to the Revised American Fertility society criteria 1985, and 15 normal controls were separated and induced by lipopolysaccharide and phytohemagglutinin, followed by 48 hour incubation in vitro, then the supernatant were collected. The levels of IL-6, IL-8 and TNF alpha were measured by enzyme linked immunosorbent assays, the concentrations of NO, expressed by nitrite and nitrate (NO2-/NO3-) content were measured by Griess methods. RESULTS: The concentrations of IL-6, IL-8, TNF-alpha and NO were significantly higher in both mild and severe Em groups than those in controls (P < 0.01). No differences between the mild and severe Em groups was seen. There were strong positive correlation between the levels of IL-6 and IL-8, TNF-alpha and NO in patients with endometriosis (P < 0.01). CONCLUSIONS: The immune function of peripheral mononuclear cell in patients with endometriosis was disordered. It may activate peripheral mononuclear cells to produce high levels of IL-6, IL-8, TNF-alpha and NO, which may take part in the pathogenesis of endometriosis. PMID- 11477802 TI - [Determination of monocyte chemotactic protein-1 in cultured endometriotic cells]. AB - OBJECTIVE: To investigate the clinical significance of monocyte chemotactic protein-1 (MCP-1) produced by endometriotic tissues. METHODS: Expressions of MCP 1 mRNA and MCP-1 protein were determined by dot blot analysis, immunohistochemical method [streptavidin biotin-peroxidase complex (SABC)] and enzyme linked immunosorbent assay (ELISA) methods in cultured endometriotic cells with median (controls) or with interleukin-1 beta(IL-1 beta) 2 ng/ml (IL-1 beta group), and tumor necrosis factor-alpha(TNF-alpha) 20 mg/ml(TNF-alpha group) respectively. The endometriotic tissues were sourced from 15 patients with endometriosis. Meanwhile, the MCP-1 protein content of cultured supernatent was also measured. RESULTS: After exposured to IL-1 beta or TNF-alpha, the expression of MCP-1 mRNA and MEP-1 protein in the endometriotic cells were significantly higher than that in the control group (P < 0.01, P < 0.05 respectively); so was the MCP-1 protein content of supernatent. CONCLUSIONS: IL-1 beta and TNF-alpha can up-regulate the expression of MCP-1 in endometriotic cells, which may be related to the development of endometriosis. PMID- 11477803 TI - [The regulation of estrogen and tamoxifen to bcl-2 oncogene in endometrial carcinoma]. AB - OBJECTIVE: To study the regulation mechanisms of bcl-2 oncogene in endometrial carcinoma. METHODS: A reverse transcription-polymerase chain reaction (RT-PCR) method was utilized to quantitate bcl-2 mRNA levels which regulated by estrogen(17-beta-estradiol) and tamoxifen in estrogen receptor (ER)(+) RL-952 endometrial carcinoma cells. RESULTS: bcl-2 mRNA level was detected in ER(+) RL 952 cells. 17-beta-estradiol stimulated a concentration-dependent increase in bcl 2 mRNA level, while tamoxifen lead to a concentration-dependent decrease in bcl-2 mRNA level. CONCLUSIONS: bcl-2 oncogene expression in ER(+) endometrial carcinoma is regulated by estrogen and antiestrogen. Estrogen may up-regulate the cell bcl 2 level to inhibit cell apoptosis, while tamoxifen may down-regulate the cell bcl 2 level and induce apoptosis. PMID- 11477804 TI - [Pregnancy of patients conceived within one year after chemotherapy for gestational trophoblastic tumor]. AB - OBJECTIVE: To explore the risk of pregnancy in patients conceived within one year after successful chemotherapy for gestational trophoblastic tumor. METHODS: 22 patients conceived within one year after chemotherapy were followed up and analysed about abnormal pregnant result, wastage rate and the time of interval between chemotherapy and pregnancy. RESULTS: Among 22 cases, 9 cases were full term birth, 6 cases were wastage. The wastage rate was 27.3%. The wastage rate of these patients conceived within half a year was higher than one year (P < 0.05). 1 repeated hydatidiform mole and one post term choriocarcinoma occurred in 22 patients. They both were conceived within 5 months after chemotherapy. CONCLUSIONS: Preservation of fertility is feasible in patients suffering from choriocarcimona and invasive mole. But these patients should practise contraception at least half a year after chemotherapy, and it's better to advise patients to take contraception for one year. PMID- 11477805 TI - [Evaluation of safety in Chinese women with amenorrhea following injection of depot-medroxyprogesterone for contraception]. AB - OBJECTIVE: To determine whether serum estradiol (E2) level was low after Depot medroxyprogesterone (DMPA) induced amenorrhea for contraception, thus to evaluate the safety of using DMPA. METHODS: Forty four Chinese women with amenorrhea after injecting DMPA were investigated, and the median period of amenorrhea was 18 months. Symptoms, pelvic examination, serum E2 and follicular stimulating hormone (FSH) levels, and vaginal cytology were examined. RESULTS: Women with amenorrhea had neither symptoms nor genital atrophy that similar to menopausal syndrome. The median level of serum E2 was 150.5 pmol/L (equal to the level of early follicular phase), and the median level of serum FSH was 14.0 IU/L. There were no correlation between E2 and FSH levels, and no correlation between amenorrhea period and E2 or FSH levels (P > 0.05) as well. No significant relationship between estrogen effects by vaginal cytology and the time period of amenorrhea was found. However, the estrogen effect was significantly correlated to the interval from last injection to vaginal smear tested. CONCLUSION: It could be concluded that though DMPA may induce amenorrhea, it will not lower serum E2 levels, and the inhibition of ovarian function is reversible. PMID- 11477806 TI - [Measurement of pregnancy-associated plasma protein A in maternal peripheral blood and Down syndrome]. PMID- 11477807 TI - [Progresses in the study of hereditary factors of polycystic ovarian syndrome]. PMID- 11477808 TI - [Clinical uses of transvaginal ultrasonography in endometrial surveillance]. PMID- 11477809 TI - [Improvement of dosage forms of traditional Chinese drugs]. PMID- 11477810 TI - [Clinical study on effects of zhiling decoction in treating senile vascular dementia]. AB - OBJECTIVE: To observe clinical effects of Zhiling Decoction (ZLD) in treating senile vascular dementia. METHODS: Seventy-eight patients were randomly divided into treated group and control group. In treated group which was treated with ZLD, a self-formulated proved recipe. In control group, which was administered with Naofukang ([symbol: see text]). RESULTS: ZLD could decrease the level of serum cholesterol, serum triglyceride and plasma LPO significantly; increase the level of high density lipoprotein (HDL) and SOD in red blood cells; dilate cerebral vessels, increase cerebral blood flow. As for P300, an index to reflect intelligence, the latency of P3 was shorter and the amplitude was raised, the brain electrical activity monitoring (BEAM) was improved remarkably, the revised Nagayakawa dementia scale scoring (NDS) of patients promoted, clinical symptom was improved significantly. Total effective rate in clinical practice reached 87%. CONCLUSION: ZLD had good effects on senile vascular dementia indeed. PMID- 11477811 TI - [Clinical study on active factors of vascular endothelial cells in acute cerebral infarction patients and therapeutical effect of activating blood stasis]. AB - OBJECTIVE: To investigate the level of plasma tissue plasminogen activator (tPA), plasminogen activator inhibitor (PAI), 6-keto-prostaglandin F1 alpha(PGF1 alpha), thromboxane B2(TXB2), factor VIII related antigen (VIII R:Ag) in acute cerebral infarction (ACI) patients with different TCM syndrome-type and the therapeutical effect of activating blood stasis (ABS). METHODS: Plasma fibrolysin system, prostaglandin system, and VIII R:Ag of 20 healthy subjects and 66 ACI patients were determined. The 45 of 66 cases were treated with ABS randomly and prospectively, 20 cases were treated by difibrase batroboxobin (DF-521), only 25 cases were treated by DF-521 together with Heart-Brain Mixture (HBM), on activating blood stasis mixture, concurrently. RESULTS: (1) The activity of tPA, ratio of tPA/(tPA + PAI) and level of PGF1 alpha decreased significantly, the level of VIII R:Ag increased remarkably in ACI patients than those of the controlled subjects. (2) Between Differentiation of Deficiency and Excess Syndromes, the level of above index changed more remarkably in the case with Excess Syndrome. Insignificant change of index shown in Deficiency cases except activity of tPA. (3) No more improvement of nerve impairment was shown between group of DF-521 together with HBM and DF-521 alone within 30 days. However, the improvement of index such as level of VIII R:Ag, ratio of tPA/(tPA + PAI) and TXB2/PGF1 alpha showed significant change between the beginning and end of treatment of DF-521 with HBM group. CONCLUSIONS: The vascular endothelial impairment was more serious in ACI patients with Excess than those with Deficiency Syndrome. DF-521 combining with HBM could protect vascular endothelial cells and improve the fibrolytic system and prostaglandin system. PMID- 11477812 TI - [Clinical study on buyang huanwu decoction to the metabolic imbalance of endothelin and calcitonin gene related peptide in patients with early cerebral infarction]. AB - OBJECTIVE: To explore the mechanism of Buyang Huanwu Decoction (BHD) in treating early cerebral infarction. METHODS: Seventy cases with early cerebral infarction were randomly divided into two groups. Chinese medicine group (CMG, n = 35) was treated with BHD; western medicine group (WMG, n = 35) was treated with hydroxyethyl starch injection and enteric coated aspirin tablets. The levels of endothelin (ET) and calcitonin gene related peptide (CGRP) in plasma before and after treatment and the results of clinical treatment were observed, and also controlled with healthy subjects. RESULTS: The levels of ET before treatment in both patient groups were significantly higher than that of the healthy subjects (P < 0.001), and the levels of CGRP were significantly lower (P < 0.001). After treatment the metabolic imbalance of ET and CGRP improved significantly in both treatment groups (in CMG P < 0.001; in WMG (P < 0.01), but the ET and CGRP in CMG improved more obviously than those in WMG (P < 0.01, P < 0.05). The marked effective and cure rate of CMG was higher than that of WMG (68.6% vs 31.4%; chi 2 = 9.65, P < 0.01). CONCLUSIONS: BHD could improve the metabolic imbalance of ET and CGRP in patients with early cerebral infarction and by this mechanism it was able to treat cerebral infarction. PMID- 11477813 TI - [Clinical study of purified xuefu capsule in treating angina pectoris]. AB - OBJECTIVE: To study the effect of purified Xuefu Capsule (PXFC) in treating angina pectoris (AP). METHODS: Fifty-seven patients with AP were randomly divided into two groups. Group A (30 cases) was treated with PXFC and western medicine, and group B(27 cases) with western medicine (isosorbide dinitrate, diltiazem or atenolol) alone. RESULTS: The total effective rates and ECG ST-T changes of AP were 93.3% and 63.3% respectively in group A. These results were all superior to those of group B (P < 0.05). Moreover, in group A the level of plasma endothelin (ET-1) decreased from 85.09 +/- 37.56 ng/L to 61.19 +/- 4.02 ng/L, and that of calcitonin gene related peptide (CGRP) increased from 58.64 +/- 19.30 ng/L to 88.87 +/- 20.41 ng/L. Comparing with group B, these changes were statistically different (P < 0.01). CONCLUSIONS: The effects of adding PXFC on conventional treatment with western medicine were better than those of western medicine. PMID- 11477814 TI - [Yishen huanshuai recipe retard progression of chronic renal failure]. AB - OBJECTIVE: To investigate the effect of Yishen Huanshuai recipe (YSHSR) on the progression of chronic renal failure (CRF). METHODS: Forty-six patients with CRF were divided into two groups: group A (self-controlled group, n = 18), whose treatment was subdivided into two stages. During the first stage, the patients treated with low protein diet and controlling blood pressure, while in the second stage, YSHSR was given. Group B(n = 28), whose treatment plan was similar with that of group A in second stage. The rate of progression of CRF was estimated by slope of the creatinine reciporcal (dl/mg) with time (month). RESULTS: Mean slope of the creatinine reciprocal with time from the group A during the first stage was -0.0104 +/- 0.0021, while during the second stage, it was -0.0034 +/- 0.0018. There was significant difference between them (P < 0.05). Mean slope from the group B was -0.0047 +/- 0.0020. There was also significant difference between that from the group B and that from the group A during the first stage (P < 0.05). CONCLUSIONS: Low protein and controlling blood pressure plus YSHSR therapy could markedly retard the rate of progression of CRF. PMID- 11477815 TI - [Effects of Chinese herbs xianzhen tablet on the deformability of erythrocyte in non-insulin-dependent diabetes mellitus patients with deficiency of both qi and yin and deficiency of kidney with blood stasis]. AB - OBJECTIVE: To explore the effects of Xianzhen Tablet (XZT) on the hemorheology of type 2 diabetes (NIDDM) on the basis of previous studies in XZT, that have indicated its ability to deplete fasting blood glucose, to reduce the injury of free radical to the body, to improve the Na(+)-K(+)-ATPase in the erythrocyte membrane of NIDDM. METHODS: Erythrocyte deformability and aggregation were studied with Ektacytometer in a total of 60 type 2 diabetes who were randomly divided into two groups. One was ZXT treated group, another was placebo group. RESULTS: (1) The erythrocyte deformability of type 2 diabetes was lower than that in normal subjects (P < 0.001), but the erythrocyte aggregation increased abnormally than that of health people (P < 0.001). The correlation test indicated there were negative correlation with plasma fibronogen and positive correlation with blood cholesterol (P < 0.05). (2) The erythrocyte deformability in the ZXT treated group with 30 patients improved after 8 weeks treatment (P < 0.01), with the decreased fasting blood glucose and plasma fibrinogen (P < 0.05). Compared with placebo group, there was significant statistical difference (P < 0.01). CONCLUSIONS: XZT could improve the erythrocyte deformability in the type 2 diabetes. PMID- 11477816 TI - [A clinical observation on the leukopenia treated with shengbaikuai decoction]. AB - OBJECTIVE: To study the effective orally taken medicine in treating the leukopenia. METHODS: Ninety cases of leukopenia induced by chemotherapy were divided into 3 groups. They were test group treated with Shengbaikuai Decoction (SBK), control group and blank group. Their efficacies were compared with each other, 25 cases with nonchemotherapy were also observed. RESULTS: WBC count and absolute neutrophil count (ANC) in test group were higher than those of other groups immediately after treatment and 2 weeks later (P < 0.01). The marked effective rate and total effective rate of test group after 2 weeks were 60.0% and 80.0% (P < 0.001) respectively, which were significantly higher than those of other groups. WBC and ANC count after treatment and 3 weeks later were higher than those before treatment in nonchemotherapy group (P < 0.001). The marked effective rate and total effective rate 3 weeks later were 64.0% and 80.0% respectively. CONCLUSIONS: SBK could increase WBC count rapidly and definitely without apparent side effect. PMID- 11477817 TI - [Clinical observation on effect of comprehensive immunotherapy in treating hepatic carcinoma after embolism chemotherapy]. AB - OBJECTIVE: To probe the relationship between survival period and immune function of hepatic carcinoma patients after embolism chemotherapy. METHODS: One hundred and eight patients of hepatocarcinoma were randomly divided into three groups. Group A was given simple embolism chemotherapy (ECT), immunotherapy and Chinese herbal medicine were given to Group B and C respectively after ECT. RESULTS: The short-term effective rate of Group A, B and C was 61.1%, 83.3% and 80.6% respectively, three-year survival rate 28.6%, 39.1% and 47.3%, median length of survival 11, 17 and 15 months respectively. The difference in terms of these indices were significant. Observation on immune function after treatment showed that patients received ECT alone had a lower immunity than those supplemented with immunotherapy or Chinese herbal medicine. CONCLUSIONS: Period of survival of hepatocarcinoma patients is related with their immune function. Patients with better immunity could enjoy a relatively extended period of survival. PMID- 11477818 TI - [Effect of kidney yin-yang replenishing Chinese herbs on the binding character of insulin receptor on hepatocyte membrane in androgen-induced sterility rats]. AB - OBJECTIVE: To observe the effect of Kidney Yin-Yang replenishing Chinese herbs on the binding character of insulin receptor (IR) on hepatocyte membrane from androgen-induced sterility rats (ASR). METHODS: The number and affinity of IR on hepatocyte membrane of ASR were determined with radioligand binding assay. RESULTS: There was no significant difference of the number or affinity for both high and low affinity sites of IR between normal and ASR. CONCLUSION: The treatment with Kidney Yin-Yang replenishing Chinese herbs had no effect on insulin binding although a relatively high rate of ovulation induction on ASR was observed. PMID- 11477819 TI - [Effect of aloe-emodin on proliferation of vascular smooth muscle cells after arterial injury]. AB - OBJECTIVE: To study the effect of Aloe-emodin (AE), an active ingredient of Rhubarb, on the kinetics of proliferation of smooth muscular cells (SMCs) cultured in vitro after rabbit iliac arterial injury. METHODS: Forty-eight hours after de-endothelialization (balloon endothelial denudation), the iliac arteries of the Japanese white rabbits were isolated and the smooth muscle cells were cultured primarily. AE was added to culture medium containing 10% fetal calf serum (FCS). The cultures were pulse-labeled with 3H-TdR and TdR uptake into VSMC were measured and the cell cycle of the cultures were analyzed by using flow cytometer. RESULTS: Compared with control, when the concentration gradient ranged from 10(-1) to 10(-5) g/L, the amount (cpm, count per minute) of 3H-TdR uptake into SMCs has significant differences (P < 0.05) and 10(-1) and 10(-2) g/L AE showed strong inhibitory effects on TdR uptake into VSMC and the percentage of inhibition [% inhibition = (cpm without AE - cpm with AE)/cpm without AE x 100%] was more than 90%. AE displayed concentration dependent inhibitory effects. The percentage of cells in G0/G1 phase was increased, but the percentage of cells in S phase was decreased in AE group, the transition of SMC cycle phase from G0 to S was blocked. CONCLUSIONS: AE is a strong inhibitor to the proliferation of SMCs and the pharmacological action of AE might reduce SMC proliferation in vivo and decrease intimal hyperplasia of restenosis. PMID- 11477820 TI - [Effects of tetramethylpyrazine and radix salviae miltiorrhizae on collagen synthesis and proliferation of cardiac fibroblasts]. AB - OBJECTIVE: To explore the effects of Tetramethylpyrazine (TMP) and Radix Salviae Miltiorrhizae (RSM) in collagen synthesis and proliferation of cardiac fibroblasts. METHODS: Using collagenase and trypsase digested rat cardiac tissue assay to isolate cardiac fibroblasts (Fbs). Different dosage of TMP, RSM and norepinephrine (NE) were used to study their effects on the collagen synthesis and proliferation of cultured cardiac Fbs. RESULTS: Compared with the control group, moderate or high dosage TMP and RSM could significantly inhibit the collagen synthesis and the proliferation of cultured cardiac Fbs. Moreover, low dose TMP(50 mg/L) and low-dose RSM(3 g/L) could antagonize the collagen synthesis and the proliferation of cultured cardiac FB stimulated by NE (500 micrograms/L). CONCLUSION: Both TMP and RSM could inhibit these processes. The mechanisms of these effect might be correlated to their Ca++ antagonistic action. PMID- 11477821 TI - [Experimental study on effect of altitude xishi capsule in treating oleic acid induced lung injury in acute hypoxia rats]. AB - OBJECTIVE: To explore the effect and pathophysiological mechanism of Altitude Xishi Capsule (XSC) on lung injury during acute hypoxia. METHODS: A new model of acute lung injury was established by administering oleic acid into superior vena cava in experimental rat during acute hypoxia, which was formed by putting the animal in a hypobaric room, and reduced the pressure gradually in uniform speed till a simulated environment of 6000 m above sea level was formed and lasted for 12 hours. XSC was given orally to the rat before oleic acid administration. The effect of XSC on the model was observed by determination of PaO2, wet/dry ratio of lung and pathologic examination at 1, 2, 3 and 4 hours separately. RESULTS: The respiratory frequency of model animal increased markedly, with the water content of lung reduced and aggravated as the hypoxia prolonged. XSC could obviously improve the pulmonary edema and inflammatory cell infiltration in lung. CONCLUSIONS: The aforesaid acute lung injury model with successful rate of 100%, it resembles the clinical acute lung injury and was useful for research on pathophysiology and treatment of ARDS or acute lung injury, XSC could improve the oleic acid induced lung injury in hypoxia rats by reducing the pulmonary edema and inflammatory cell infiltration. PMID- 11477823 TI - [Early clinical education on integrated traditional Chinese and Western medicine]. PMID- 11477822 TI - [Blocking action of tetrandrine on the scar-collagen matrix contraction stimulated by transforming growth factor beta]. AB - OBJECTIVE: To investigate the effect of Tetrandrine on the scar-collagen matrix contraction stimulated by transforming growth factor beta (TGF-beta). METHODS: The scar-derived fibroblasts were suspended in a solution of collagen matrix, and TGF-beta, they were associated to form a scar model in vitro. The influence of tetrandrine on the contraction indexes was observed. RESULTS: Tetrandrine plays a blocking action in the scar-collagen matrix contraction stimulated by TGF-beta, which was in a dose-dependent manner. CONCLUSION: This function of tetrandrine could be responsible for the anti-scar mechanism. PMID- 11477824 TI - [Advances in the study of anti-inflammatory and immunoregulatory effects of saikosaponins and their similar substances]. PMID- 11477825 TI - [Clinical and experimental study of effect on acute phase protein level of multiple organ dysfunction syndrome treated with dachengqi decoction]. AB - OBJECTIVE: To investigate the level of acute phase protein level (APP) synthesized in hepatocytes in pathogenic process of multiple organ dysfunction syndrome (MODS) and the effect of Dachengqi Decoction (DCQD) on it. METHODS: The levels of C-reactive protein (CRP), alpha 1-acid glycoprotein (alpha 1-AGP), alpha 1-antitrypsin total (alpha 1-AT), alpha 2-macroglobulin (alpha 2-MG), ceruloplasmine (Cp), hepatoglobin (Hp) and transferrin (Tf) synthesis were determined in 32 cases of MODS patients and rat models of MODS induced by acute infective peritonitis and mesenterial artery ischemia-reperfusion. RESULTS: The serum APP level in MODS patients was increased and higher than that in the control group (P < 0.01). After 3 days of DCQD treatment, it reduced significantly (P < 0.05). Serum APP level in MODS model was also increased significantly, and after 3 days of DCQD treatment, it reduced significantly (P < 0.05). The APP secreting level of rat hepatocytes cultured in vitro raised after stimulated and activated by lipopolysacchride, which was reduced after cultured with rabbits' serum containing different concentration of effective ingredients of DCQD. The APP inhibiting effect of DCQD was dose dependent. CONCLUSIONS: Purgation therapy with DCQD could reduce the stimulation of pathogenic factors on hepatocytes, protect the tissues and organs from injury caused by excessive inflammatory reaction. PMID- 11477826 TI - [Clinical study on relationship among Heart-Qi Deficiency, left ventricular function and myocardial ischemia in patients with coronary heart disease]. AB - OBJECTIVE: To study the relationship among Heart-Qi Deficiency (HQD), left ventricular function and myocardial ischemia in patients with coronary heart disease (CHD). METHODS: Doppler echocardiogram, electrocardiogram were used to observe 103 patients of CHD clinically. Patients of stable angina pectoris treated with Radix Codonopsitis Pilosulae (50 cases) or Nifedipine (20 cases) were observed as well. RESULTS: Occurrence of left ventricular function insufficiency, both systolic and diastolic, was higher in CHD patients of HQD type as compared with that of non-HQD type. The left ventricular diastolic function showed high susceptibility (87%) and the left ventricular systolic function showed high specificity (88%) for HQD evaluation. Radix Codonopsitis Pilosulae could protect the left ventricular function and improve myocardial ischemia. CONCLUSIONS: Myocardial ischemia and blood stasis cause HQD, while the Chinese herbal medicine treatment on HQD could improve the blood stasis to alleviate myocardial ischemia effectively. The mechanism of which might be to enhance the positive diastolic and systolic function of cardiac muscles, reduce the resistance pressure and keep pressure of coronary perfusion so as to improve the blood supply of heart. PMID- 11477827 TI - [Effect of dauricine on redistribution of glycoprotein IV in platelet membrane of patients with mitral stenosis]. AB - OBJECTIVE: To investigate the effect of dauricine on the irreversible platelet aggregability of patients with mitral stenosis (MS). METHODS: Glycoprotein IV (GP IV) and thrombospondin (TSP) levels on the membrane surface of the stationary platelet or platelet activated by thrombin (0.05 U/ml, 0.1 U/ml, 0.5 U/ml, 1.0 U/ml) in 16 patients with MS were measured with flow cytometric method and compared with those of the healthy (14 cases). RESULTS: The GP IV level of stationary platelet, the GP IV and TSP levels of activated platelet in MS patients were higher than those in the healthy significantly (P < 0.01), while the TSP level of stationary platelet was not different between the patients and the healthy (P > 0.05). The GP IV redistribution on the activated platelet surface was apparently inhibited by dauricine (50 mumol/L, P < 0.05, P < 0.01) and the release of TSP from intracellular alpha-granules was inhibited by dauricine only in the activated platelets induced by thrombin of low concentration (0.05 U/ml and 0.1 U/ml, P < 0.05, P < 0.01), inhibiting effect was not found in those activated with high concentration of thrombin. CONCLUSION: The activity and reactivity to thrombin of platelets increased in MS patients, and dauricine was able to reduce the occurrence of the irreversible platelet aggregation in MS patients. PMID- 11477828 TI - [Study on effect of acupoint sticker of TTS-ST93-1 in treating motion sickness]. AB - OBJECTIVE: To observe the effect of Transdermal Therapeutic System ST93-1 (TTS ST93-1) by acupoint sticking in treating motion sickness (MS). METHODS: One hundred and sixty-three young volunteers with history of MS recently were subjected to a 3 hours journey by track and armored car, and divided randomly into tested, positive control and negative control groups, treated with TTS-ST93 1 plaster, TTS-Scopolamine plaster and placebo (lactose plaster) respectively. The TTS-ST93-1 plaster was stuck on Neiguan (P6) or Shenque (CV8) or Qimai (TE18) point and the effect of treatment was observed by double-blind method. RESULTS: The mean effective rate of tested group was 95.9%, which was significantly different to that of the negative control group (P < 0.01). In the tested group, the protection rate of plaster stuck on Shenque was 81.4%, on Neiguan was 45.5% and on Qimai was 100%, with an average of 76.0%, which was significantly different to that of the negative control group. The protection rate of TTS Scopolamine plaster was merely 27.3%. The side effects of TTS-ST93-1 plaster were mouth dryness, sleepiness and fatigue, as compared with the negative control group, the difference was insignificant. No allergic reaction of skin such as swelling, pain or itching was found in the tested group. CONCLUSION: Acupoint sticker of TTS-ST93-1 was an effective treatment for MS with less side effect and no allergic reaction of skin. PMID- 11477829 TI - [Clinical study on qiangxinling liquid extract in treating congestive heart failure]. AB - OBJECTIVE: To find the safe and effective treatment and drugs for congestive heart failure (CHF). METHODS: Ninety CHF patients whose heart function belong to III-IV grade were randomly divided into 3 groups: Group A was treated with Qiangxinling liquid extract (QXLLE); Group B with QXLLE plus small dose of digoxin and Group C with digoxin alone. RESULTS: The total effective rate on heart function of Group A was 86.7%, Group B 93.3% and Group C 83.3%, the effect of Group B was better than that of the other two groups (P < 0.05, P < 0.01). The digoxin withdrawing and reducing rate in Group B was higher than that in Group C (86.7% vs 50.0%, P < 0.05). The left ventricular systolic function, including stroke volume, cardiac output, ejection fraction of the 3 groups were improved after treatment, among them the effect of Group B was the best (P < 0.01). Moreover, improvement in ventricular diastolic function was shown in Group A and B, including the maximum blood flow of the late and the early diastolic stage as well as the ratio of them (P < 0.05, P < 0.01). The plasma renin activity, angiotensin II and atrial natriuretic peptide of Group A and B were also lowered after treatment and were significantly different to those of the Group C (P < 0.01). CONCLUSIONS: QXLLE could improve the heart function, clinical symptoms and neuro-endocrinal indexes of CHF patients and reduce the side effects of digoxin. PMID- 11477830 TI - [Clinical and experimental study on treatment of acute cerebral infarction with Acanthopanax Injection]. AB - OBJECTIVE: To evaluate the effect of Radix Acanthopanacis Senticosi in treating acute cerebral infarction and its mechanism. METHODS: Clinical study using single blind, control trial, in which 60-80 ml of Acanthopanax Injection (AI) added to 500 ml of 0.9% normal saline was given by intravenous drip once a day for 14 days in the AI group(34 patients), while 500 ml of dextra-40 was given by the same way in the control group(26 patients). In experimental study, rabbit model of acute incomplete cerebral ischemia was made by ligation of bilateral common carotid artery. 10 ml of AI were given once daily intravenously for 7 days to the AI group and 10 ml of normal saline was given to the control group in the same way. Clinical effects (in clinical study) and effect of AI on cerebral oxygen free radicals (OFR), superoxide dismutase (SOD), and serum concentration of ACTH and cortisone were observed (in experimental study). RESULTS: Clinical study showed that the total effective rate of AI group (86%) was higher than that of the control group (50%) while nervous functional deficit score was lower in AI group (9.96 +/- 4.66) than that in the control group (13.56 +/- 1.84) significantly. Experimental study showed that the cerebral level of OFR decreased while SOD increased and serum concentration of ACTH and cortisone decreased after treatment in AI group. CONCLUSIONS: AI was effective in treating acute cerebral infarction, the mechanism is probably by ameliorating peroxidation in brain and improving hypothalamic-pituitary-adrenocortical axis function. PMID- 11477831 TI - [Effect of chon-ren regulating herbal medicine on hormone level in patients of mammary hyperplasia]. PMID- 11477832 TI - [159 sterility patients of yang-deficiency of spleen and kidney treated by shouwu huanjing capsule]. PMID- 11477833 TI - [Experimental study on treatment of viral myocarditis in mice by integrated traditional Chinese and Western Medicine]. AB - OBJECTIVE: To clarify the effects of Astragalus Membranaceus (AM) combined with taurine and/or coenzyme Q10(CoQ10) on coxsackievirus B3 (CVB3) murine myocarditis. METHODS: Viral myocarditis model was created by intraperitoneal inoculation with CVB3 solution and were treated by saline, AM, taurine, CoQ10, AM + taurine, AM + CoQ10, AM + taurine + CoQ10, respectively. The mortality, ECG, CVB3-RNA in myocardium and myocardial histopathologic changes were observed. RESULTS: AM combined with taurine and CoQ10 could significantly reduce the mortality of the mice and the incidence of abnormal ECG at acute stage. CVB3-RNA was significantly reduced in AM treated group, especially in AM + taurine group. No anti-virus effect was found in CoQ10 group. All drugs could lighten myocardial histopathologic changes and the effect could be enhanced by combined treatment. CONCLUSIONS: AM, taurine and CoQ10 have some curative effects on CVB3 murine myocarditis, AM combined with taurine and CoQ10 is the best. PMID- 11477834 TI - [Effect of qingjieling induced endogenous granulocyte-colony stimulating factor in strengthening host defence against bacteria]. AB - OBJECTIVE: To explore the effect of Qingjieling (QJL) induced endogenous granulocyte-colony stimulating factor (G-CSF) in strengthening host defence against bacteria in mice. METHODS: Serum containing G-CSF was obtained by injecting 5 mg/g of QJL to mice. The CSF activity was identified by ELISA, bone marrow cell proliferation and colony-forming assays. Forty-eight hours after the serum was infused to 8 normal BALB/C mice, the mice was sacrificed and the number of E. coli in spleen suspension was measured. Eight mice of the control group were treated in the same way but with normal serum of mice instead of the QJL treated serum. RESULTS: The QJL treated serum showed a positive reaction with anti-G-CSF McAb, it promoted the proliferation of bone marrow cells and formation of cell colony of neutrophilic granulocyte type in the culture medium, these results indicated that the serum contained G-CSF undoubtedly. The number of E. coli in spleen suspension of test group was significantly lower than that of the control group P < 0.01). CONCLUSIONS: QJL does enhance the defense capability of the host, the mechanism of which partly might be due to the induction of endogenous G-CSF in the host. PMID- 11477835 TI - [Experimental study on reducing effect of shexiang baoxin pill from damage of arterial wall caused by hyperlipemia]. AB - OBJECTIVE: To study the protecting effect of Shexiang Baoxin Pill (SXBXP) on arterial wall in hyperlipemia rabbit model. METHODS: Thirty-six male, New Zealand rabbits were randomized into three groups: control group, fed with common diet for 8 weeks: Hypercholesterolemia group (HC) and SXBXP group (SX), fed with 1% cholesterol and 3% lard diet for 8 weeks. In SX group, SXBXP was iven from 7th week for 2 weeks. The plasma levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were measured at the first, 6th and the 8th week of experiment. Serum superoxide dismutase (SOD) and electrocardiography (ECG) were also evaluated at the 8th week. Light microscopy and transmission electron microscopy were adopted to assess the degree of pathologic damage of arterial wall. RESULTS: SXBXP could reduce the levels of serum TC and LDL-C, as compared with the HC group, P < 0.05. It could also reduce the extent of atherosclerotic lesion and the severity of intimal proliferation. CONCLUSION: SXBXP has the effect of inhibiting development of experimental atherosclerosis. PMID- 11477836 TI - [Exploration on mechanism of Chinese medical recipe for cough-remedy and sputum removing]. PMID- 11477837 TI - [Progress in diabetic retinopathy treated by integrated Chinese and Western Medicine]. PMID- 11477838 TI - [Treatment of leukemia with traditional Chinese drugs containing arsenic]. PMID- 11477839 TI - [Clinical study on effect of concentrated xuefu zhuyu pill on restenosis of 43 cases coronary heart disease after intracoronary stenting]. AB - OBJECTIVE: To explore the clinical effectiveness of concentrated Xuefu Zhuyu Pill (CXZP) on restenosis (RS) of coronary heart disease (CHD) after intracoronary stenting. METHODS: Eighty-four successful intracoronary stenting patients were randomly divided into conventional treatment group (control group, 41 cases) and conventional treatment plus CXZP group (treated group, 43 cases). The follow-up underwent for 6 months. Clinical symptom, change of blood stasis were observed. And occurrence of restenosis was estimated by ECG exercise treadmill testing, emission computed tomography (ECT) and coronary angiography (CAG). RESULTS: The symptoms of blood stasis of treated group improved in the follow-up significantly (P < 0.01), and the integral of blood stasis were decreased significantly in the treated group vs the conventional treatment group (P < 0.01). The recurrent angina pectoris rate of the treated group was lower than the conventional treatment group (P < 0.01). The 61.9% of 84 patients reexamined the CAG in the follow up, and the lower restenosis rate (25.8%) in the treated group than in the conventional treatment group (32.1%, P = 0.59). CONCLUSION: There is the trend that CXZP could prevent restenosis. Therefore, it is necessary to increase the cases for further study of CXZP on preventing restenosis. PMID- 11477840 TI - [Clinical study of jiuxin fumai injection in treatment of cardiogenic shock]. AB - OBJECTIVE: To increase the cure and improvement rates and to decrease the mortality rate in the treatment of cardiogenic shock. METHODS: Thirty-one patients with cardiogenic shock were studied, and received Jiuxin Fumai injection (JXFM) in research group (n = 20) or dopamine (DA) in control group (n = 11). RESULTS: Eighteen effective cases (90.0%) and 2 ineffective cases (10.0%) in JXFM group, 9 effective cases (81.8%) and 2 ineffective cases (18.2%) in DA group. There was significant difference statistically, P > 0.05. CONCLUSIONS: JXFM and DA had the same effect in elevating blood pressure, enhancing pump function of the heart and ameliorating collapse syndrome. They are safe, effective, convenient for use and prompt in responding. However, JXFM showed raising blood pressure faster, higher as well as more stable, and the dual phase phenomenon in adjusting heart rate, which could slow down tachycardia as well as speed up bradycardia were observed. PMID- 11477841 TI - [Clinical observation of yugengongyu decoction in treatment of coronary heart disease with angina pectoris]. AB - OBJECTIVE: To observe the clinical effect of Yugengtongyu Decoction (YGTY) in treatment of coronary heart disease (CHD) with Qi deficiency, Blood stasis and phlegm stagnancy type and Qi deficiency and Blood stasis type. METHODS: Patients were divided into two groups, 45 cases in the YGTY group and 38 cases in the control group. In alleviating angina pectoris and improving symptom of CHD, the serum total cholesterol (TC), serum triglyceride (TG) were determined before and after treatment. RESULTS: The total effective rate of spontaneous angina pectoris were significantly better in the treated group than those in the control group (P < 0.005). In improving symptom of CHD were significantly better in the treated group than those in the control group (P < 0.005), and it was better the type of Qi deficiency, Blood stasis and phlegm stagnancy than type of Qi deficiency and Blood stasis in treated group (P < 0.05). TC of the treated group was lowered markedly after treatment, as compared with that before treatment, the difference was significant (P < 0.005). CONCLUSIONS: YGTY was effective for alleviating angina pectoris and improving symptom of CHD. It is best for the type of Qi deficiency, Blood stasis and phlegm stagnancy. It also has regulatory effect on the blood lipid. PMID- 11477842 TI - [Salvia Miltiorrhizae Composita in preventing PaO2 lowering induced by controlled hypotension with nitroglycerin]. AB - OBJECTIVE: To observe the effects of Salvia Miltiorrhizae Composita (SMC) on blood gas variations of nitroglycerin (NTG) controlled hypotension. METHODS: Sixteen patients who were arranged to undergo operation under general anesthesia in controlled hypotension condition were divided randomly into control group (n = 8) and SMC group (treated group, n = 8), NTG were used to create controlled hypotension in both groups and blood pressure decrease about 30% comparing with the control values. Patients in the treated group were administered intravenously with SMC (16 ml) before the utilization of NTG. RESULTS: Significant PaO2 decrease and Pa-etCO2 increase were observed in the control group during the controlled hypotension period. However, blood gas values in the treated group did not evidently change. CONCLUSION: It suggested that SMC can prevent the side effects of NTG on blood gas. PMID- 11477843 TI - [Clinical study on Shuanghuanglian powder in treating children viral myocarditis]. AB - OBJECTIVE: To study immune function of children viral myocarditis and to evaluate the clinical effect of Shuanghuanglian Powder (SHLP). METHODS: The authors determined serum COXB-IgM and the numbers of lymphocyte subsets CD3+, CD4+, CD8+, CD4+/CD8+ with viral myocarditis patients by ELISA and indirect immunofluorescent assay. Sixty-two cases were divided randomly into two groups. Thirty-two cases treated by conventional therapy with SHLP and the other 30 cases treated with conventional therapy alone were taken as control group. RESULTS: COXB-IgM was positive in 39 of 62 patients, which were significantly different with those of normal controls (P < 0.001). In addition, the level of CD4+ cells and the ratio of CD4+/CD8+ were decreased while CD8+ increased. After treatment with SHLP, the recovery of symptoms, signs and immune function in patients were better than that of controls. These changes were significantly different (P < 0.01). CONCLUSIONS: There were disturbance of immune regulatory function with children viral myocarditis patients and SHLP is an effective drug in treating children viral myocarditis. PMID- 11477844 TI - [Clinical study on treatment of aplastic anemia mainly with Yishen Shengxue tablet]. AB - OBJECTIVE: To assess the clinical effects of Yishen Shengxue tablet (YSSXT) on aplastic anemia. METHODS: All patients were divided into two groups by simple randomized methods, 106 patients of the treatment group were treated with YSSXT, 36 patients of the control groups were treated with Renshen Guipi Pill (RSGPP). Peripheral blood and bone marrow of the two groups of patients were measured before and after treatment. The toxicity and side effects of YSSXT were also observed. RESULTS: The basical cure rate, remission rate, total effective rate of the treatment and control groups were 23.6%, 36.8%, 82.1% and 11.1%, 22.2%, 58.3% respectively. There was significant difference between the two groups (P < 0.05 and P < 0.001). The effect of treatment group was obviously better than that of control group. Follow up study showed that from onset of the disease to follow up time in the treatment group, the longest one was 18 years 4 months, so the long term curative effect is promising. During the period of clinical observation, no obvious side effect and toxicity of YSSXT were found. CONCLUSION: YSSXT is an effective and safe drug in treating aplastic anemia. It has a good prospect and application. PMID- 11477845 TI - [Clinical and experimental study on treatment of acute alcohol intoxication with xiangnaojing injection]. AB - OBJECTIVE: To study the therapeutical mechanism of traditional Chinese medicine Xingnaojing injection (XNJI) for acute alcohol intoxication. METHODS: XNJI was used in treating the experimental model rabbits (n = 26) and the patients (n = 8) admitted to the emergency department with acute alcoholism. Before and after the treatment, beta-EP, superoxide anion (free radicals) and SOD were measured. RESULTS: XNJI could enhance the regaining consciousness of rabbits and patients, simultaneously reduce the concentration of beta-EP in plasma to the normal level (drunk rabbits 127.09 +/- 13.67 ng/L, normal rabbits 41.48 +/- 7.46 ng/L. P < 0.01, drunk patients 292.97 +/- 14.85 ng/L, normal people 221.60 +/- 15.95 ng/L, P < 0.01). The concentration change of superoxide anion (free radicals) in plasma of rabbits and patients was similar to beta-EP (drunk rabbits 313.39 +/- 15.64 u/L, normal rabbits 254.27 +/- 21.71 u/L, P < 0.01; drunk patients 278.47 +/- 11.48 u/L, normal people 159.92 +/- 11.51 u/L, P < 0.01), and SOD was inversely changed (drunk rabbits 53.57 +/- 6.48%, normal rabbits 77.18 +/- 7.89%, P < 0.01; drunk patients 43.76 +/- 7.84%, normal people 82.53 +/- 4.33%, P < 0.01). CONCLUSIONS: XNJI is similar to Naloxone in pharmacologic action. And it is an effective antioxidant. It can be used for treating alcoholism. PMID- 11477846 TI - [Clinical observation of forty cases in adolescent dysmenorrhea treated by traditional Chinese medicine]. AB - OBJECTIVE: To observe the therapeutic effect of Tongjing powder (TP) and understand the change in pathophysiology of dysmenorrhea, the blood flow parameters of uterine arteries and hemorheological data before and after the TP administration. METHODS: The blood of both uterine arteries of patients was examined by color doppler flow imaging. The hemorheological data was examined by conical viscosimeter. The two groups were compared with each other. RESULTS: Pulsation index (PI), resistance index (RI), systolic peak value/diastolic peak value (A/B value) of dysmenorrhea patients and hemorheological data except erythrocyte sedimentation rate of dysmenorrhea patients were significantly higher than the normal control group. After treating with TP, no difference existed between two groups. CONCLUSIONS: The blood of dysmenorrhea patients appear to be viscous and adhesive, TP can improve the state of blood circulation and treat the dysmenorrhea effectively. PMID- 11477847 TI - [Effect of Fusheng powder on activity of vascular endothelial cells and its adhesion to polymorphonuclear neutrophils]. AB - OBJECTIVE: To explore the effect of Fusheng powder on the adhesion of polymorphonuclear neutrophils (PMN) in human peripheral blood to vascular endothelial cells (VEC). METHODS: Using cultured human umbilical vein cells as target cells, while being incubated with high glucose (HG, 30 mmol/L), high-lipid (HL, 20%), tumor necrosis factor-alpha (TNF-alpha, 10%) and hypoxia (95% N2, 5% CO2, 37 degrees C, 30 min, and then in the air 30 min) for 24 h, respectively, the activity of VEC and the adhesive effect of PMN to VEC were surveyed. RESULTS: It revealed that HG, HL, TNF-alpha and hypoxia could enhance obviously the adhesion of PMN to VEC, the cell attachment rate in normal control, HG group, TNF alpha, hypoxia and HL after 30 minutes contact were 100%, 129.6%, 136.7%, 151.4% and 167.7% (all P < 0.01, compared with control group) respectively, but high and low dosage of the Fusheng powder could significantly inhibit the adhesion of PMN to VEC. CONCLUSIONS: HG, HL, TNF-alpha and hypoxia could all activate EC respectively and enhance the adhesion of PMN to VEC, but the Fusheng powder could protect VEC and partly block the effect of this adhesion. However, the effects of Fusheng powder on anti-adhesion are of great importance in clinical cardiovascular and cerebral vascular diseases. PMID- 11477848 TI - [Experimental study of Yishou tiaozhi tablet on inhibiting hyperlipemia and atherosclerosis]. AB - OBJECTIVE: To observe the effect of Yishou Tiaozhi tablet on lipid-metabolism and aortic intimal atherosclerotic plaque coverage in rabbit's model of experimental hyperlipemia and atherosclerosis. METHODS: Thirty-two rabbits were divided randomly into four groups, group A, B, C and D, 8 in each group. Forage with cholesterol and lipid plus 1.59 g/kg of Yishou Tiaozhi tablet was fed to the group A every day; to the group B, 22.54 mg/kg gypenoside tablet was added to forage with cholesterol and lipid; to the group C, forage with cholesterol and lipid was fed; and to the group D, ordinary forage only. Biochemical parameters were measured and pathomorphological examination were carried out 6 weeks later. RESULTS: (1) Yishou Tiaozhi tablet obviously lowered the levels of serum TC, TG, AI, ApoB, LP (a), ox-LDL, HYP, plasma Ca2+, TXB2 and increased the levels of serum HDL-ch, ApoA1, ApoA1/APoB, plasma 6-keto-PGF1 alpha(P < 0.01). (2) Pathomorphological examination showed that in the group A aortic intimal atherosclerotic plaque area and arterial intima thickness obviously reduced, smooth muscle cell hyperplasia and elastic fibers were not seen. CONCLUSION: Yishou Tiaozhi tablet can inhibit experimental hyperlipemia and atherogenesis. It is an ideal and effective medicine in preventing and treating hyperlipemia and atherosclerosis. PMID- 11477849 TI - [Effect of Yangxue bushen tablet on ovarian function in animal model of Yang deficiency]. AB - OBJECTIVE: To study the therapeutic mechanism of Yangxue Bushen tablet (tablet for nourishing blood and tonifying Kidney) on the Kidney deficiency type of functional amenorrhea and infrequent menstruation. METHODS: Applied with orimeten, models of Yang deficiency in female rabbits were made, ikaclomine was taken as the control drug. Group I was the group of animal model of Yang deficiency; Group II was the ikaclomine treatment group; Group III was the high dosage treatment group; Group IV was the low dosage group; Group V normal rabbit treated with distilled water. The functional effect of gonadal axis was evaluated by the changes of the signs of animal histomorphology of ovary and uterus, and the level of beta-endorphin. RESULTS: The animals of Yang deficiency in Group II, III and IV recovered as a whole, and the changes of different levels of folliculi, morphosis of endometria and deciduous vaginal epithelial cell in group II, III and IV were more obviously than those in group I and V (P < 0.05). The content of beta-endorphin in blood plasma in group I was 106.6 pg/ml, which was lower than those in the other groups (P < 0.05), and the contents of beta endorphin in blood plasma, in group III, IV and II were similar to that in group V (P < 0.05). CONCLUSION: Yangxue Bushen tablet has the effect on regulating menstruation by regulating the function of gonadal axis, regulating and promoting the ovarian function. PMID- 11477850 TI - [Experimental study on preventive effect of traditional Chinese medicine replenishing recipe on acute oxygen toxicity caused by hyperbaric oxygen]. AB - OBJECTIVE: To seek for the effective traditional Chinese medicine (TCM) replenishing recipe (RR) prescription that being able to prevent the onset of oxygen convulsion and to offer some clues in preventing the oxygen convulsion with TCM-RR. METHODS: Mice were administered orally with four respective representative RR, Sijunzi Decoction, Siwu Decoction, Liuwei Dihuang Decoction, and Renshen Dihuang Decoction. Then the convulsion latent periods were recorded and SOD vitalities in brain and lung were determined. RESULTS: Compared with the control groups, the convulsion latent period of Sijunzi Decoction group was prolonged significantly (P < 0.01) and its SOD vitality in lung was maintained higher (P < 0.01) after convulsion, while the other three RR results have no significant difference (P < 0.05). CONCLUSION: Sijunzi Decoction has certain preventive effect on acute oxygen convulsion. PMID- 11477851 TI - [Progress in the treatment of epilepsy in children with traditional Chinese medicine]. PMID- 11477852 TI - [Pharmacokinetics of Chinese herbal drug combinations]. PMID- 11477854 TI - [The traditional Chinese medicine culture and modern quality control]. PMID- 11477853 TI - [Integrated traditional Chinese medicine with western medicine: carrying forward the advantages and avoiding the shortcoming to develop jointly]. PMID- 11477855 TI - [Observation of clinical effect of acupuncture on vascular dementia and its influence on superoxide dismutase, lipid peroxide and nitric oxide]. AB - OBJECTIVE: To observe the clinical effect of electroacupuncture and acupuncture on vascular dementia (VaD) and the influence on superoxide dismutase (SOD), lipid peroxide (LPO) and nitric oxide (NO) levels. METHODS: Forty-six cases of VaD were randomly divided into two groups, the electroacupuncture group (EA group) and the acupuncture group (AP group). Assessment of Hasgawa's dementia scale (HDS), functional activities questionnaire (FAQ) and neurologic deficit scoring were done before and after treatment, and the changes on SOD, LPO and NO levels were observed. RESULTS: After treatment, in the EA group, the HDS elevated, the FAQ lowered, SOD increased and LPO and NO decreased significantly, as compared with before treatment, P < 0.01. But these parameters were not changed significantly in the AP group after treatment. The clinical symptoms were improved in both groups, but the reduction on neurologic deficit score was not significant. The total effective rate of the EA group was higher than that of the AP group. CONCLUSION: The immediate effect of electroacupuncture was superior to that of acupuncture, suggesting that the electroacupuncture was more effective in promoting the intelligence recovery than acupuncture. PMID- 11477856 TI - [Curative effect of TCM-WM therapy on Wilson disease with different clinical phenotypes and polymorphisms of ATP7B gene]. AB - OBJECTIVE: To investigate the curative effect of TCM-WM therapy on Wilson disease (WD) with different clinical phenotypes and polymorphisms of ATP7B gene. METHODS: One hundred and twenty-two WD patients with different clinical phenotypes were treated with unithiol 20 mg/kg per day and Gandou tablet ([symbol: see text]) for 1 month. Exon 18 of ATP7B gene extracted from patients and 20 healthy volunteers were amplified with PCR technique, mutations and polymorphisms were screened by SSCP analysis. RESULTS: Four kinds of abnormal migration bands in PCR-SSCP were seen in 37 patients, mutation frequencies of three different clinical phenotypes and curative effect between mutation and non-mutation patients did not show any significant difference, P > 0.05. But total effective rate of patients with Wilson type or pseudosclerosis type was significantly higher than that of patients with hepatic type, P < 0.05. CONCLUSIONS: The exon 18 of ATP7B gene of Chinese WD patients may be one of the regions with higher mutation frequency, most of them are compound heterozygotes and patients with different clinical phenotypes have different response to TCM-WM therapy. It indicate that specific mutations at least in part play a role in influencing clinical phenotypes and curative effect. PMID- 11477857 TI - [Studies on plasma norepinephrine and epinephrine contents and vegetative nervous function in liver syndrome]. AB - OBJECTIVE: To study the characteristics of plasma norepinephrine (NE) and epinephrine (E) contents alteration and vegetative nervous function disturbances in patients with Liver Syndrome of TCM. METHODS: Subjects were selected by combination of disease diagnosis and Syndrome Differentiation. Plasma NE and E contents were determined by high performance liquid chromatography (HPLC) and electrochemical detector. The vegetative nervous function were assessed by comprehensive analysis of sympathetic and parasympathetic hyperfunction symptoms, physiologic indexes and physical loading tests. RESULTS: Patients of Liver Excess Syndrome had an elevated plasma NE and E contents and with the vegetative nervous system disturbance mainly hypersympathetic. On the contrary, patients of Liver Deficiency Syndrome had an lowered plasma NE and E contents and with the vegetative nervous system disturbance mainly hyperparasympathetic. CONCLUSION: Plasma NE and E contents and vegetative nervous function disturbance can be taken as the auxiliary laboratory diagnostic indexes for Syndrome Differentiation of Liver Excess and Liver Deficiency. PMID- 11477858 TI - [Clinical study on Shenqi Fuzheng injection combined with chemotherapy in treating malignant tumor of digestive tract]. AB - OBJECTIVE: To assess the clinical effect and safety of Shenqi Fuzheng Injection (SQFZI) combined with chemotherapy in treating malignant tumor of digestive tract. METHODS: One hundred and eighty-two patients were randomly divided into 3 groups, the combined therapy group (Group A), the SQFZI group (Group B) and the chemotherapy group (Group C). The remission and stabilization effect on cancer were observed. RESULTS: The remission rate and stabilizing rate of Group A were 20.2% and 87.9%, and those of the Group C were 15.6% and 68.9%, the difference between the two groups was significant, P < 0.05. The symptom and living quality improving rate of Group A were 78.8% and 42.4% separately, those of the Group B were 73.7% and 68.4% and of the Group C 37.8% and 26.7% respectively, the inter group differences were significant, P < 0.05. The weight gain rate and effective rate of Group A were 45.5% and 90.9%, of Group C were 13.3% and 40.0%, P < 0.05. SQFZI showed good protective effect on hematopoietic system, 7.7% patients in Group A with WBC count lower than 4 x 10(9)/L after treatment, while in Group C, the percentage reached 22.2%. Moreover, SQFZI could raise activity of NK cell, macrophage and T-lymphcyte subgroups, without any injury on heart, liver and kidney function or other adverse reaction. CONCLUSION: SQFZI combined with chemotherapy has certain effect of remitting and stablizing on digestive tract tumor, the clinical effect is significant to patients with Qi-Deficiency, and is safe and reliable. PMID- 11477859 TI - [Effects of traditional Chinese medicine on salivary glands in the patients with head and neck cancer during radiotherapy]. AB - OBJECTIVE: To observe the effect of Chinese herbal medicine on the salivary glands of patients with head and neck cancer during radiotherapy (RT). METHODS: Fifty cases with head and neck cancer were randomly divided into observation group treated with RT plus Chinese herbal medicine and control group treated with RT alone. The level of mouth dryness, the flow and secretion speed of salivary amylase were measured before and during the RT. RESULTS: The severity of mouth dryness increased and the flow and secretion speed of salivary amylase declined during RT. The mouth dryness curve of patients in Chinese medicine group was significantly more flattened than that of the control group after RT. The flow and the secretion speed of salivary amylase of the observation group were significantly higher than those of the control group at 20 Gy, 40 Gy and 60 Gy. CONCLUSION: Chinese herbal medicine used in this study was effective to relieve the severity of mouth dryness and to protect the salivary glands of patients with head and neck cancer during RT. PMID- 11477860 TI - [Immune modulatory and therapeutic effect of lentinan on condyloma acuminatum]. AB - OBJECTIVE: To observe the immune modulatory and therapeutic effect of lentinan on condyloma acuminatum (CA). METHODS: Thirty-six CA patients were randomly divided into two groups, 19 in the test group treated with lentinan plus CO2 laser irradiation and 17 in the control group treated with laser irradiation alone. Their T-lymphocyte subsets of peripheral blood and level of serum interleukin-2 (IL-2) and soluable interleukin-2 receptor (SIL-2R) were determined before and after treatment, and the recurrence rates of the two groups were compared. RESULTS: After treatment, in the test group, the CD4/CD8 ratio, serum IL-2 raised and serum SIL-2R lowered significantly (P < 0.05, 0.05, 0.05 respectively) as compared with before treatment, while those parameters were not changed significantly in the control group. The recurrence rate of the test group was lower than that of the control group, P < 0.05. CONCLUSION: Lentinan could modulate the cellular immunofunction of CA patients and reduce the recurrence rate of CA. PMID- 11477861 TI - [Clinical and experimental study of Tongqiao tablet in treating catarrhal otitis media]. AB - OBJECTIVE: To assess the effect of Tongqiao tablet (TQT) in treating catarrhal otitis media (COM). METHODS: Ninety-six patients of COM were randomized into two groups, 52 patients in the treatment group were treated with TQT and 44 patients of the control group were treated with western medicine. Animal experiment study on effect of TQT was also carried out. RESULTS: The total effective rate and the cure and markedly improved rate of the treatment group were 92.31% and 53.85% respectively, while those in the control group were 68.18% and 27.27% respectively, the difference between the two groups was significant statistically (P < 0.01). The effect of TQT was superior to that of the western medicine. Experimental study indicated that TQT had remarkable inhibiting effect on inflammation and granuloma formation. CONCLUSION: TQT is an effective drug for treatment of COM. PMID- 11477862 TI - [Follow-up study on influence of Sini Decoction to living quality of patients after percutaneous transluminal coronary angioplasty]. AB - OBJECTIVE: To determine the influence of Sini Decoction (SND) on living quality of patients after percutaneous transluminal coronary angioplasty (PTCA). METHODS: Randomized case-control clinical trail was conducted to evaluate the living quality of 40 post-PTCA patients before and after SND treatment by scoring. RESULTS: The scores in physical symptoms, sense of well-being, depression, satisfaction of life and work capacity of patients after PTCA were improved significantly as compared with before PTCA, P < 0.05-0.01. Scores of patients received SND treatment were higher than those untreated with SND in the first three criteria, P < 0.05, particularly on relieving palpitation and dyspnea. CONCLUSION: SND is helpful to improve the living quality of patients after PTCA. PMID- 11477863 TI - [Treatment of diabetes mellitus by integrated traditional Chinese and western medicine]. PMID- 11477864 TI - [Relationship between endothelin-1 and ischemic brain damage after subarachnoid hemorrhage and protective effect of Ginkgo biloba extract]. AB - OBJECTIVE: To investigate the role of endothelin-1 (ET-1) in development of ischemic brain damage after subarachnoid hemorrhage (SAH), and the protective effect of Ginkgo biloba extract (GBE). METHODS: Wistar rat noncraniotomy models of SAH were divided into SAH group and GBE treated group, the diameter of basilar artery (BA) and dynamic changes of regional cerebral blood flow (rCBF) and ET-1 content of intracranial plasma within 24 hours after SAH of both groups were determined. And pathological examination of CA1 region of hippocampus was performed 3 days later. RESULTS: rCBF decreased and ET-1 content increased obviously and retained in 24 hours after SAH. Spasm of BA occurred half an hour after SAH and neurons of hippocampus CA1 region was damaged severely. GBE could antagonize the above-mentioned pathological changes effectively. CONCLUSION: Increase of ET-1 is an important factor leading to ischemic brain damage after SAH. GBE exerts its protective effect by antagonizing pathological increase of ET 1. PMID- 11477865 TI - [Experimental study of vascular smooth muscle cell apoptosis induced by Huotong Capsule]. AB - OBJECTIVE: To explore the anti-atherogenic mechanism of Huotong Capsule (HTC) on vascular smooth muscle cell (VSMC) apoptosis. METHODS: VSMC proliferation model of cultured human embryonal thoracic aorta was established by stimulating VSMC with interleukin-1 (IL-1) and HTC was applied to observe the apoptosis inducing effect by transmission electron microscopy, DNA gel electrophoresis and flow cytometry. RESULTS: Typical apoptotic cellular changes were found after HTC treatment, the VSMC shrinked, the nuclei disappeared after chromatin agglutination, cellular membrane projected and wrapped the organelle to form apoptotic body. DNA gel electrophoresis showed "ladder" strand of DNA, a special phenomenon of cellular apoptosis. And an Ap peak of degraded DNA could be seen by flow cytometry, which widened and heightened when the concentration of HTC increased. The above-mentioned changes were not found in the blank control and untreated model group. CONCLUSION: HTC could induce apoptosis of human proliferated VSMC. PMID- 11477866 TI - [Study on effect of Erigeron injection in prevention and treatment of cerebral ischemic injury]. AB - OBJECTIVE: To study the effect of Erigeron injection (EI) on cerebral ischemic injury. METHODS: Animal model of brain ischemic/reperfusion was established in male Wistar rats and the effect of EI on proteinkinase C (PKC) activity, intraneuronal free calcium and regional cortex blood flow (rCBF) were observed. RESULTS: EI could prevent the activation of PKC, calcium overload and decrease of RCBF induced by cerebral ischemia/reperfusion. CONCLUSION: EI could prevent neuronal ischemic injury by inhibiting activation of PKC. PMID- 11477867 TI - [Prospect on over early stage treatment of ischemia apoplexy with traditional Chinese medicine]. PMID- 11477868 TI - [Current status of the study on the effects of traditional Chinese drugs on cardiovascular disease related receptors]. PMID- 11477869 TI - [Progress in the study of immuno-pharmacology of yupingfeng powder]. PMID- 11477870 TI - [Physicians of the respiratory tract disease specialty should take the lead and be the example in the control of smoking]. PMID- 11477871 TI - [The relationship between different serological types of Pseudomonas aeruginosa strain in lower respiratory tract and its clinical significance]. AB - OBJECTIVE: To explore the relationship between different serological types of pseudomonas aeruginose (PA) in sputum and its clinical significance. METHOD: PA was detected by Vitek-Ams GNI card and its sensitivity to 12 kinds of antibiotics was measured with GNS-GA, its different serological types were assayed with standard diagnostic serum of type 20. RESULT: 14 different serological types were isolated from 101 strains of PA in sputum (with the typed rate 84.2%). Among them, type 6, type 1, and type 3 were higher with 23.7%, 13.9% and 10.9% respectively. But another 15.8% of strains couldn't be differentiated, so called untyped PA group, among them 10 strains with mucoid-like colony (63% of this untyped group). Patients suffered with this group were older and were very seriously infected with higher mortality (13%). They often suffered from serious COPD and bronchiectasis. The sensitivities of all PA strains to antibiotics in vitro as follows: tobramycin 92.0%, ceftazidine 89.1%, ciprofloxacin 87.1%, ticarcillin 82.2%, and gentamycin 76.2%. Among 12 PA strains tested with imipenem sensitive test, only one strain of mucoid-like colony had been found resistant. CONCLUSION: Identification of serological type of PA can be used as an important method to make out if the infection comes from the same source, and for surveillance of hospital infection, and to show the main types of PA in some district. But the sensitivity of antibiotics in vitro is no definitive correlation with PA serological types. The disease infected with untyped PA strains have its clinically special characteristics and must be kept attention. PMID- 11477872 TI - [Distribution and drug-resistance of 345 strains gram-negative bacilli from hospitalized patients with lower respiratory tract infections diseases]. AB - OBJECTIVE: To investigate the distribution and drug-resistance of gram-negative bacilli in respiratory ward. METHOD: (1) Drug-resistance was tested by Kirby Bauer disk sensitivity method and MIC by agar doubling dilution. (2) beta lactanase was tested by nitrocephin. (3) ESBLs was tested by E test. RESULT: (1) Among 345 strains gram-negative bacillus, klebsiella pneumoniae and pseudomonas aeruginoea was about 39.7%, 17.4%, respectively. (2) Drug-resistance rate of cefotaxime dropped from 42.9% (6 years ago) to 8.2%. (3) beta-lactamase positive rate of 345 strains bacilli was 54.5%. (4) ESBLs was negative. CONCLUSION: (1) Klebsiella pneumoniae and pseudomonas aerugries were the main pathogens in lower respiratory tract infection in hospital. (2) Although we didn't find the evidence of ESBLs, we should pay more attention to it in the future. PMID- 11477873 TI - [A clinical study of six cases of Pneumocystis carinii pneumonia]. AB - OBJECTIVE: To heighten the awareness of pneumocystis carinii pneumonia. METHOD: Six cases of pneumocystis carinii pneumonia were retrospectively analyzed. RESULT: The underlying diseases were leukemia, lymphoma, renal transplantation and autoimmune hemolytic anemia. The clinical features were dyspnea, cough and fever. X rays typically showed a bilateral diffuse or patchy interstitial infiltrate. Three cases complicated with acute respiratory failure showed hypoxemia, intrapulmonary shunt and reduced compliance resembling ARDS. CPAP/PEEP was required. High-level PEEP (> 1.47 kPa) can be applied to the refractory hypoxemia. SMZco and dapsone were the initial choice of treatment. CONCLUSION: The prognosis of pneumocystis carinii pneumonia with acute respiratory failure was poor. Aggressive early diagnosis and treatment were critical to improve survival. PMID- 11477874 TI - [Carcinosarcoma of the lung clinical analysis]. AB - OBJECTIVE: To improve the diagnosis and treatment of pulmonary carcinosarcoma. METHOD: Two patients with pulmonary carcinosarcoma were confirmed by operation and pathology were presented and relevant literatures were reviewed. The histo pathological, clinical features, diagnosis, differential diagnosis and treatment were introduced appearance. RESULT: Carcinosarcoma of the lung is rare. It is 0.2%-0.6% of pulmonary malignant tumor. Carcinosarcoma of the lung may be divided into 2 types: First, the endbronchial type which tends to grow slowly in a polypoid pattern along the major bronchi with local invasive feature. The second, arising peripherally which tends to metastasize early with serious outcome. CONCLUSION: This tumor may be misdiagnosed as brondrogeuic carciniose preoperatively. It tends to metastasize early and prognosis is poor. Surgical intervention is the first choice. PMID- 11477875 TI - [Effect of tetrandrine on the collagen contents of lungs in rats with chronic hypoxia]. AB - OBJECTIVE: To investigate the effect of tetrandrine (Tet) on the collagen contents of lung tissues in rats with chronic hypoxic pulmonary hypertension. METHOD: Using colorimetric method after oxidized by chloramine T to determine hydroxyproline contents of lung tissues and extrapulmonary arteries, using Masson method, image pattern analysis technique and gray scale scanning to observe the changes of collagen distribution in extrapulmonary arteries and lung tissues in rats with hypoxia and the effect of treatment with Tet. RESULT: Tet could remarkably reduce the mean pulmonary arterial pressure and pulmonary vascular resistance in rats with hypoxic pulmonary hypertension and could inhibit the increase of collagen contents in lung tissues and extrapulmonary arterial wall. CONCLUSION: Tet may be used as one of the main drugs in the treatment of chronic hypoxic pulmonary hypertension due to its action of reducing mPAP and collagen contents. PMID- 11477876 TI - [Changes of the deformability of neutrophil and the influence of anisodamine on neutrophil deformability during endotoxin-induced acute lung injury in rats]. AB - OBJECTIVE: To explore the mechanisms of change of deformability of polymorphonuclear leukocytes (PMN) and influence of anisodamine on deformability of PMN during endotoxin-induced acute lung injury (ALI) in rat. To explore the role of PMN in ALI. METHOD: The model of ALI in rat was reproduced by intravenous injection of E Coli endotoxin. The deformability of PMN was measured by micropipette aspiration technique. RESULT: The elastic modulus K1 of PMN at the 1st hour (33 +/- 12 Pa), 4th hours (37 +/- 12 Pa, 1 Pa = 0.0075 mmHg) and 6th hours (38 +/- 13 Pa) after injection of E Coli endotoxin are all significantly higher than that in the control group (26 +/- 9 Pa P < 0.01). The viscosity modulus (mu) of PMN at the 4th hours (5 +/- 3 Pa.s) and 6th hours (6 +/- 3 Pa.s) after injection of E Coli endotoxin are all significantly higher than the control group (3.1 +/- 2.6 Pa.s, P < 0.01). The elastic shear modulus (G) of PMN at the 4th hours (54 +/- 20 Pa) and the 6th hours (56 +/- 21 Pa) after injection of E Coli endotoxin are all significantly higher than the control group (47 +/- 17 Pa, P < 0.05). The K1(33 +/- 12 Pa) and mu(4 +/- 3 Pa.s) in anisodamine group are all significantly lower than endotoxin group in the same time phases (P < 0.05). CONCLUSION: The deformability of PMN decrease remarkably during ALI, which will enhance PMN sequestration in lungs, lead to ALI. It may be one of important pathogenic mechanism of ALI. The anisodamine can inhibit the decrease of deformability of PMN effectively and may have some therapeutic effect on ALI. PMID- 11477877 TI - [Investigation and analysis of the diagnostic status of bronchial asthma]. AB - OBJECTIVE: The aim of this study was to investigate the rate of delayed diagnosis of the bronchial asthma and related factors. METHOD: 57 asthmatic outpatients were investigated randomly by questionnaire including main symptoms of asthmatic attack, the date of initial attack, the date of initial visit physician, and initial diagnosis, the basis of diagnosis and the names of misdiagnosed diseases. Then according to the above-mentioned data the duration of delayed diagnosis, the rate of delayed diagnosis were calculated. RESULT: 96% of outpatients saw physician within 1 yr. after initial attack. The duration from initial visit to making diagnosis were 11.1 +/- 2.3, and 1.5 +/- 0.3 yr. before and after 1980, respectively, and their rate of delayed diagnosis were 75%, 15%, respectively. Both of their differences were statistically significant (P < 0.01). Only 20% of diagnosis of asthma was by pulmonary function test. CONCLUSION: The majority of the delayed diagnosis should be due to making the mis-understanding and underattention of physician to the asthma, and their diagnostic levels should need further improving. PMID- 11477878 TI - [CD4+ T cell activation and IL-5 production in atopic and nonatopic asthmatics]. AB - OBJECTIVE: To understand the reasons and roles of CD4+ T cell activation and IL-5 production in atopic and asthmatic patients. METHOD: Bronchoalveolar lavage(BAL) cells and peripheral blood mononuclear cells (PBMC) from 12 atopic and 10 nonatopic asthmatics, 9 atopic nonasthmatics, and 10 normal controls were cultured with or without house dust mite(HDM), CD4+ T cell activation, and IL-5 production were assessed. RESULT: The percentage of CD4+, CD25+ in unstimulated PBMC cultures were not significantly different in the four groups, but it increased following HDM stimulation in both PBMC and BAL cultures in two atopic groups(P < 0.01). The percentage of CD4+, CD25+ in unstimulated BAL cell culture increased in two asthmatic groups (P < 0.05, P < 0.01). The levels of spontaneous IL-5 released from both BALF cells and PBMC in two asthmatic groups were higher than those in AN and N (P < 0.05, P < 0.01). A significant elevation in IL-5 release following HDM stimulation in PBMC and BALF cells was observed in the both asthmatic and atopic groups; but the value in AA was higher than that in NAA, and it was also higher in BALF cells than in PBMC. CONCLUSION: The allergen stimulation is important reasons for CD4+ T cell activation and IL-5 production in atopics. CD4+ T cell activation and IL-5 production are common feature in atopic and nonatopic asthma, it correlates with both asthmatic and atopic status. PMID- 11477879 TI - [The clinical diagnostic significance on detection of Mycoplasma pneumoniae antigen in upper respiratory tract epithelium]. AB - OBJECTIVE: To investigate the early diagnosis of pulmonary mycoplasma pneumoniae (MP) infection by detection of MP antigen in upper respiratory tract epithelium. METHOD: A prospective study on respiratory MP infection was conducted in 110 patients with acute respiratory infection from February 1996 to November 1997 in our hospital. Upper respiratory tract epithelia were collected from gargle and MP antigen was detected by immunofluorescent assay (IFA). At the same time, serum samples were taken and immunoglobulin M (IgM) fractions of specific antibody to MP were assessed by the same method in 70 of 110 patients. Moreover, clinical features, cold agglutination test and chest X-ray were also collected for analysis. RESULT: The positive rates of MP antigens and IgM antibody were 17% and 20%, respectively, while the titer on cold agglutination test > 1:8 in only two patients. The clinical features in patients with positive MP antigen and/or IgM antibody were similar to that of pulmonary MP infection. CONCLUSION: Examination of MP antigen in upper respiratory tract epithelium collected from gargle combined with serum specific IgM antibody is a good method for the early diagnosis of pulmonary MP infection. PMID- 11477880 TI - [Comparing the bronchodilating effects of ipratropium bromide and theophylline]. AB - OBJECTIVE: To compare the bronchodilating effects of ipratropium bromide and theophylline in chronic obstructive pulmonary disease(COPD). METHOD: In 26 patients, mean peak forced expiratory volume in 1 second(FEV1) were measured at the time before and 30 minute, 1, 2, 3, 4, 5, 6 hours after the drugs were administered in a double-blind, placebo-controlled crossover method. RESULT: The mean peak of FEV1, increased over baseline were 34% and 19% respectively, for ipratropium bromide and theophylline (P < 0.01), and the reaching duration were 1 2 hours and 2-3 hours, the proportion of patients attaining at least a 15% (P < 0.01) increase in the FEV1, responders were 19% and 50% (P < 0.01) respectively, for ipratropium bromide and theophylline. The average FEV1, increased during the 6-hours observation period were 18% for ipratropium bromide and 8% for theophylline (P < 0.01). The mean duration of FEV1 > 15% was 3.6 and 1.6 hours respectively. CONCLUSION: Ipratropium bromide nebulized is a more potent bronchodilator than the oral theophylline in patients with chronic airflow obstruction. PMID- 11477881 TI - [The changes of EG1 and EG2 positive eosinophils and their clinical significance in asthmatics]. AB - OBJECTIVE: To evaluate the significance of EG1 or EG2 positive eosinophils in relating to the severity of asthma. METHOD: EDTA-anticoagulated venous blood were treated by FOG method, and FTTC-conjugated monoclonal antibodies were added. Flow cytometry technique was used. RESULT: EG1 and EG2 positive eosinophils increased significantly in patients with acute asthma (n = 29) as compared with control (n = 13) and COPD (n = 12), P < 0.05. There were significant correlation of the mean fluorescence intensity(MFI) between EG1, EG2 and the degree of the symptom. CONCLUSION: There is a significant increase of activated eosinophils in peripheral blood in asthmatics, the expression of EG1 and EG2 positive eosinophils correlated well with the severity of symptom in asthmatics. EG1 and EG2, as the marker for eosinophils activity, may be sensitive and specific in evaluating the severity of asthmatics symptom. PMID- 11477882 TI - [Culture of mycobacterium tuberculosis of gastric juice collected through different ways in diagnosis of pulmonary tuberculosis in children]. AB - OBJECTIVE: To explore the value of culture of Mycobacterium tuberculosis through gastric juice specimens in diagnosis of patients with pulmonary tuberculosis in children. METHOD: Sputum, early morning vomited gastric juice (MVG), the other time vomited gastric juice (OVG) and early morning gastric lavage (MGL) were collected from 46 children with suspected pulmonary tuberculosis. All the specimens were processed to culture for Mycobacterium tuberculosis using pyruvate media, and the results were compared. RESULT: Of the 46 cases, Mycobacterium tuberculosis was found in 4 sputum samples (9%), 15 MGL samples (33%), 12 MVG samples (26%) and 2 OVG samples (4%). CONCLUSION: Collecting gastric juice through different methods for culture of Mycobacterium tuberculosis is specific for diagnosis of paediatric pulmonary tuberculosis. PMID- 11477883 TI - [Comparison of three antituberculous drugs in serum and cold abscesses of patients with spinal tuberculosis]. AB - OBJECTIVE: Pharmacokinetics of INH, RFP and OFLX in serum and cold abscesses of patients with spinal tuberculosis was analyzed to provide reference to choosing clinical therapeutic regimen. METHOD: The aspiration specimens of abscesses and venous blood were collected from 8 patients with spinal tuberculosis at 0.5, 0.75, 1.0, 1.5, 2.0, 4.0, 6.0, 9.0, 12.0, 16.0 and 24.0 h after administration of antituberculous drugs. The specimens were assessed by high performance liquid chromatography (HPLC). The data were processed with software 3P87. RESULT: The Cmax of INH, RFP and OFLX in serum were 10.87 +/- 7.09 micrograms/ml, 9.98 +/- 3.53 micrograms/ml, and 5.29 +/- 0.72 micrograms/ml, while the Cmax of INH, RFP and OFLX in cold abscesses were 2.84 +/- 1.63 micrograms/ml, 0.57 +/- 0.26 microgram/ml and 3.19 +/- 1.29 micrograms/ml respectively. CONCLUSION: After administration, the Cmax of INH and OFLX in the cold abscesses of patients with spinal tuberculosis, reached the level beyond their MIC and appeared and disappeared more slowly than that in the serum; RFP was not easy to permeate into the cold abscesses, the Cmax of RFP in the cold abscesses just reached its MIC. PMID- 11477884 TI - [Clinical features, diagnosis and treatment of solitary vertebral arch tuberculosis]. AB - OBJECTIVE: To explore clinical features of vertebral arch tuberculosis, and to discuss the diagnosis and treatment of this illness. METHOD: Seventeen of 3,825 cases of spinal tuberculosis who were hospitalized and treated from 1956 to 1996 were reviewed, and their clinical features, diagnosis and treatment were analyzed and reported. RESULT: Sixteen cases of vertebral arch tuberculosis were in adolescence and prime of life, and another one was a 2-year-old child. Their symptoms in early stages were untypical, including local fixed pain and tenderness, and later, abscess, fistula and symptoms of nerve system appeared. All the cases were diagnosed through summarizing of their history, signs, imaging examination and bacteriology tests, and only one case was misdiagnosed. In one of 17 cases, antituberculosis chemotherapy was conducted alone and non-operation was performed. In the other 16 cases, focal debridement of vertebral arch was performed on the basis of antituberculous chemotherapy, and in 4 cases of them with symptoms of nerve system, laminectomy and spinal decompression was performed homochronously. All the cases were healed and no relapse occurred during follow up. CONCLUSION: It is difficult to diagnose vertebral arch tuberculosis in early stages due to its occult symptoms, but the symptom of confined pain and manifestation in X-ray, CT and MRI are much useful for early diagnosis. Abscess puncture and acid-fast staining smear microscopy can usually be used to confirm diagnosis. And surgical intervention is an important measure for its treatment. PMID- 11477885 TI - [Apoptosis of eosinophils and anti-apoptosis in asthma]. PMID- 11477886 TI - [Anti-tubercular agent injuries to the liver in liver disease]. PMID- 11477887 TI - [Controlled clinical study on efficacy of fixed-dose compounds rifater/rifinah in antituberculous chemotherapy]. AB - OBJECTIVE: To assess antituberculous efficacy, patients' compliance and application perspective of fixed-dose compounds rifater/rifinah in China. METHOD: Three hundred eight new smear positive pulmonary tuberculosis patients were randomly allocated with a ratio of 2 to 1 into treatment group (227 cases, receiving 2RIFATER/4RIFINAH regimen) and controls (81 cases, 2HRZ/4HR) for observation. RESULT: The sputum negative conversion rates at the 2nd month in the treatment group and the controls were 91.2% and 86.4% respectively, and at the end of the chemotherapy 98.7% and 97.5%. Chest radiography showed remarkable improvement. The resolution of pulmonary lesions in the treatment group and the controls accounted for 95.2% and 93.8% respectively, with cavity closure rates of 68.6% in the treatment group and 67.9% in the controls. The drug adverse reaction rates were 8.9% in both groups, and the default rates were 4.3% and 7.8% respectively. CONCLUSION: Fixed-dose compounds rifater/rifinah show excellent therapeutic efficacy, safety and compliance in antituberculous chemotherapy, which could be recommended for wider use in tuberculosis control in China. PMID- 11477888 TI - [Efficacy of regimens containing INH, RFP with varied chemotherapy courses on retreated culture positive pneumoconio-tuberculosis]. AB - OBJECTIVE: To evaluate the short and long-term effects of regimens containing INH, RFP with varied chemotherapy courses on culture-positive pneumoconio tuberculosis. METHOD: 79 patients with culture-positive pneumoconio-tuberculosis were divided into three groups according to chemotherapy duration: 9-months group (M9: 2SHRZ/7HRE) 28 cases, 12-month group (2SHRZ/10HRE) 25 cases, 18-month group (M18: 2SHRZ/10HRE/6HR) 26 cases. Evaluating the efficacy of regimens depended predominantly on sputum bacteriological conversion, and the patients who completed the regimens were followed up for 5-8 years. RESULT: Sputum negative conversion rates of three groups at the end of chemotherapy were 83%, 96%, 95%, and their recurrent rates in follow-up period 41%, 4% and 5% respectively. Of all patients who completed the regimens bacteriological relapse rates from the first to fourth year are 6%, 8%, 2%, 2% in the follow-up period. There was no bacteriological relapse from fifth to eighth year. CONCLUSION: It is effective for SHRZ/HRE combination with 12-months course to retreated tubercle bacillus positive pneumoconio-tuberculosis, and it is appropriate for such patients to be followed up for 4-5 years. PMID- 11477889 TI - [Diagnosis and treatment of tuberculous appendicitis]. AB - OBJECTIVE: To study the diagnosis and treatment of tuberculous appendicitis. METHOD: 12 cases (0.26%) with tuberculous appendicitis diagnosed by histopathological examination selected from 4,652 patients during 1968 to 1997 were analysed retrospectively. RESULT: The mean age of the patients with tuberculous appendicitis was 35 years, the disease occurred 2 times more frequently in women than in men and was usually secondary to tuberculosis elsewhere in the body (7 cases). There were 7 cases with proliferative lesions, 3 cases with ulcerative lesions and 2 cases with two kinds of lesions. 12 cases were all misdiagnosed preoperatively, 2 cases were diagnosed definitely in the operation. In 12 cases, 7 cases underwent simple appendectomy, 2 cases partial celectomy and 1 case right hemicolectomy, appendectomy and resection of regional lymph nodes were performed on 2 cases, treatment of anti-TB was given postoperatively to 9 patients. All the patients recovered without complications. CONCLUSION: It's hard to get definite diagnosis of tuberculous appendicitis before operation because of low incidence and non-specific clinical manifestations, so careful observation in the operation and routine histopathological examination must be emphasized. Early operation and postoperative treatment of anti-TB should be advocated in order to prevent from complications. PMID- 11477890 TI - [The occurrence of hypertension in patients with obstructive sleep apnea syndrome]. AB - OBJECTIVE: To determine the changes in blood pressure (BP) and its correlated factors in patients with obstructive sleep apnea syndrome(OSAS). METHOD: Retaining manometric tube in radial artery and polysomnography(PSG) were done synchronously in 13 patients with OSAS in whom was diagnosed by whole-night PSG. Oxygen therapy was given to two of the patients and NCPAP treatment was given to four of the patients on the following night. RESULT: (1) Six of the patients (46 percent) had daytime hypertension (SBP > 18.7 kPa and/or DBP > 12 kPa), the incidence of transient hypertension during apnea in 7 patients who do not have daytime hypertension was 86% (6/7). (2) Lowest oxygen saturation during sleep was significantly correlated with the level of SBP(r = -0.659, P < 0.05) and DBP(r = 0.677, P < 0.01) in awake, the baseline of SBP(r = -0.777, P < 0.01) and DBP (r = -0.590, P < 0.05) in NREM sleep and that of SBP(r = -0.585, P < 0.05) in REM sleep. In REM sleep, the changes in MBP showed significantly correlation with the changes in oxygen saturation during apnea (r = 0.598, P < 0.05). In NREM sleep, the changes in SBP were significantly correlated with apnea length(r = 0.580, P < 0.05). The changes in BP showed no correlation with apnea index in different sleep stages (P > 0.05). (3) Nocturnal elevation in BP disappeared in four patients who received NCPAP treatment, but not in two patients who received oxygen therapy. CONCLUSION: The transient hypertension during apnea might be recurrent episodes in patients with OSAS who have normal daytime BP. Nocturnal hypoxemia and apnea length might play a role in nocturnal elevation in BP in patients with OSAS. Hypoxemia was not the major factor in BP elevation after obstructive apneas. Simple oxygen administration could not normalize the elevation in BP during apnea in patients with OSAS and NCPAP might be an appropriate therapy. PMID- 11477891 TI - [Analysis of etiological agents and death-relating factors of pulmonary infections in immunocompromised patients]. AB - OBJECTIVE: To explore the distribution of etiological agents and death-relating factors of pulmonary infections in immunocompromised hosts(ICH). METHOD: 80 patients of pulmonary infections in ICH confirmed by etiological and(or) pathological diagnosis were retrospectively analyzed. RESULT: The distribution of etiological agents of pulmonary infections (except for cytomegalovirus, CMV): common bacteria 56%, mycobacterium tuberculosis 29%, fungus 11%, pneumocystis carinii 4%. Mortality of pulmonary infections in ICH showed significant difference between age 67% (> or = 60 years): 48% (< 60 years), bacteremia 86% (yes): 45% (no), white blood cell count 67%, 87% (> 10 x 10(9)/L, < 4.0 x 10(9)/L): 23% (4.0-10) x 10(9)/L, extent of lesions in x-ray film 65% (diffuse): 23% (local) (P < 0.05, P < 0.01), however, there was no obvious difference in sex, underlying diseases, types of etiological agents, mixed infections. CONCLUSION: Bacteria was one of the most common etiological agents of pulmonary infections in ICH, the latent reactivation and relapse of pulmonary tuberculosis in ICH should be emphasized sufficiently. The preliminary analysis of death relating factors showed: age, bacteremia, white blood cell count, extent of lesions in x-ray film were related to the prognosis of pulmonary infections in ICH. PMID- 11477892 TI - [Studies on the biological and clinical characteristics of acquired pneumonia caused by K. planticola]. AB - OBJECTIVE: To investigate the characteristics of biological, and clinical epidemiology of acquired pneumonia caused by a new type of Klebsiella, K. planticola. METHOD: 9 strains of K. planticola were isolated from respiratory samples of patients, in vitro, biological and serologic identification were done, in vivo, infected animal models were also evaluated. Clinical epidemiological inquiries were also performed. RESULT: 9 strains' biological features were different from those of the other Klebsiella. The 9 patients' ages were over forty and the primary disease was the third type tuberculosis of lungs. Using of antituberculosis drugs for long term as well as penicillins might be the predisposing factors, corticosteroids were used in 3 of the 9 patients, Combined infection with other pathogens in addition to K. planticola occurred in 8 patients, in 7/8, Candida albicans was identified. Drug sensitive tests show that all of them were resistant to penicillins and sensitive to the second and third generation cephalosporins. CONCLUSION: The new strain of Klebsiella was studied and this information will be useful for diagnosing and treating K. platicola. PMID- 11477893 TI - [The role of glucocorticosteroid and theophylline in asthmatic inflammation of murine model and the inhibition in NO production in lung]. AB - OBJECTIVE: To investigate the role of glucocorticosteroid and theophylline in treatment of asthmatic inflammation and their effects in production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS). METHOD: A murine model was set up (A group: asthma model, N group: control mice), then the number of WBC, eosinophils% (EOS), total protein level and the NO level in brochoalveolar lavage fluid (BALF) were measured after treatment with glucocorticosteroid (C1 group: dexamethason inhalation; C2 group: dexamethason peritoneal injection and C3 group: budisonide inhalation) and aminophylline(T group). The expression of iNOS was studied by histochemistry and immunohistochemistry. RESULT: There was a significant increase in number of WBC [(10.64 +/- 0.75) x 10(7)/L vs (3.15 +/- 0.92) x 10(7)/L, P < 0.001), EOS% (12.55% +/- 7.64% vs 0.02% +/- 0.04%, P < 0.001), total protein level (0.124 +/- 0.080 g/L vs 0.008 +/- 0.004 g/L, P < 0.001) and concentration of NO (5.00 +/- 1.87 mumol/L vs 1.30 +/- 0.67 mumol/L, P < 0.001) in BALF after antigen challenge, and the expression of iNOS was increased in airway epithelial cells and alveolar macrophages. After administration of steroids and theophylline, the number of WBC[C1: (5.05 +/- 1.14) x 10(7)/L, C2: (5.47 +/- 0.59) x 10(7)/L, C3: (6.53 +/- 0.47) x 10(7)/L, T: (3.65% +/- 1.04) x 10(7)/L, P < 0.001], EOS% (C1: 0.10% +/- 0.06%, C2: 0.10% +/- 0.08%, C3: 0.24% +/- 0.33%, T: 2.84% +/- 1.90%, P < 0.001-0.005), total protein level (C1: 0.016 +/- 0.011 g/L, C2: 0.010 +/- 0.008 g/L, C3: 0.020 +/- 0.012 g/L, T: 0.012 +/- 0.008 g/L, P < 0.001-0.05) were significantly decreased. There was a significant reduction of NO level (C1: 1.20 +/- 0.97 mumol/L, P < 0.001) and iNOS expression after treated with steroids, but no significant change after management with theophylline. CONCLUSION: EOS is main inflammatory cell in asthma. NO may play an important role in aggravating asthma. Steroids and theophylline both can inhibit cell infiltration and serum protein leakage in asthma. Steroids can also inhibit the production of NO and iNOS expression. But theophylline has no significant effect in NO. PMID- 11477894 TI - [Study on the pathological features of airway inflammation in asthma and chronic bronchitis]. AB - OBJECTIVE: To explore the difference between the pathological features of asthma and chronic bronchitis on airway inflammation, then to provide pathomorphological basis for appropriate diagnosis and treatment of these two different entities. METHOD: To compare the pathological changes of airways from 14 asthmatic and 13 chronic bronchitic patients. RESULT: There were damaging lesions shown on tracheo bronchial epithelium of asthmatic and chronic bronchitic patients; various inflammatory cells and structural pathological changes occurred in submucosa. The numbers of eosinophilic cells and mast cells in asthmatic group were higher than those of chronic bronchitic group (P < 0.01). Thickness of basement membrane in early stage (P < 0.05) and thickness of smooth muscle were characteristic pathological changes for asthmatic group. Degranulation phenomenon of mast cells was found in asthmatic group. The pathological features of airway mucosa of chronic bronchitic group were different: eosinophilic cells < 1.2/HP and mast cells < 1/HP; no degranulation of mast cells was observed under electronmicroscopy. CONCLUSION: Both asthma and chronic bronchitis (including chronic asthmatic bronchitis) are chronic airway inflammatory illnesses, but obvious pathomorphological difference was found in their airway mucosa. These are two different diseases and the reasons for causing airflow obstruction are also different. PMID- 11477895 TI - [Effects of dexamethasone on apoptosis of airway inflammatory cells in asthmatic guinea-pigs]. AB - OBJECTIVE: Clearance of airway inflammatory cells is the key point of therapeutic effect in asthma. Apoptosis, a form of cell death, is thought to be critically important to promote the clearance of inflammatory cells and the resolution of inflammation. To investigate the effects of dexamethasone on apoptosis of airway inflammatory cells in asthma showed be very important. METHOD: Dexamethasone was used for the treatment of asthma model of guinea pigs set up by inhaling ovalbumin. Apoptotic cells were labelled with TdT-mediated dUTP nick end labeling(TUNEL) technique on formalin-fixed paraffin-embedded trachea and lung tissue sections. By way of immunohistochemistry, eosinophils were stained with EG2 antibody and T lymphocytes with CD3+, CD4+ and CD8+ antibodies on sections. RESULT: (1) The quantity of airway eosinophils and lymphocytes decreased in dexamethasone group, which could not be observed in the control one (P < 0.01). (2) Apoptotic index of lymphocytes was significantly elevated following dexamethasone treatment(P < 0.05). But no difference was found in the proportion of apoptotic eosinophils between these two groups. (3) The percentage of EG2 positive eosinophils and CD4+ positive lymphocytes decreased significantly following dexamethasone treatment. On the contrary, the number of CD8+ lymphocytes increased in dexamethasone group when compared with the control one (P < 0.05). CONCLUSION: (1) The quantity of airway eosinophils decreases following dexamethasone treatment in guinea-pigs asthma, and its mechanism may not be ascribed mainly to the apoptosis of eosinophils. (2) The number of lymphocytes, mainly CD4+, decreased following dexamethasone treatment in guinea pig airways, and apoptosis may represent the mechanism that promotes the clearance of lymphocytes in asthma. PMID- 11477896 TI - [Diagnosis and treatment of pulmonary sequestration]. AB - OBJECTIVE: To explore the cause, classification, incidence, diagnosis and treatment of pulmonary sequestration. METHOD: 42 cases analysed and literatures reviewed. RESULT: 42 cases of sequestration were presents which were resected and confirmed by pathology in our department from 1964 to 1997. 29 cases (69%) were confirmed before operation. The incidence rate was 0.32% comparing with the sametime operations. There were 37 cases (88%) of intralobar type, and 5 cases (12%) of extralobar type. 22 cases accompanied with infection, 9 cases with hemoptysis in intralobar type. 1 case with infection and 1 case with hemoptysis in extralobar type, in the latter case, there was a fistula between the affected lung and the esophagus. CONCLUSION: Pulmonary sequestration is not a rare illness. The diagnosis mainly depends on X-ray. Sometimes, bronchography, retrograde arteriography or CT can also make the diagnosis correctly. Special attention should be paid to avoiding injure to the vessels which supply the isolated lung during operation. PMID- 11477897 TI - [Change of hymodynamics and circulating endothelial cells in chronic obstructive pulmonary diseases (COPD) and cor pulmonale patients]. AB - OBJECTIVE: To investigate the vascular endothelial injuries by observing the changes of circulating endothelial cells (CEC) numbers in blood and hemodynamics in COPD and cor pulmonale patients. METHOD: The arterial catheterion technique and CEC collection technique were used, superoxide dismutase (SOD) and malondialdyhyde(MDA) levels were determined respectively. RESULT: mPAP (3.76 +/- 0.75 kPa) and CEC (16.70 +/- 2.65/0.9 microliter), MDA (6.98 +/- 1.47 mumol/L) in the chronic cor pulmonale are higher than that of mPAP (2.09 +/- 0.41 kPa) and CEC (8.48 +/- 2.23 microliters) in COPD, the numbers of CEC are adversely correlated to PaO2 (gamma = 0.9423, P < 0.001) and positively correlated with mPAP (gamma = 0.8270, P < 0.001), superoxide dismutase(SOD) was decreased. CONCLUSION: mPAP, CEC are higher in cor pulmonale than that in COPD, CEC numbers rise when the PaO2 was lowered, hypoxia may worsen the vascular endothelial injuries. PMID- 11477898 TI - [Inhibition of potassium channel by chronic hypoxia on pulmonary artery smooth muscle cells in rats]. AB - OBJECTIVE: To explore the possible effect of potassium channel in chronic hypoxic pulmonary hypertension. METHOD: Male Wistar rats were placed in the identical normobaric or hypoxic environmental chamber. In one chamber, rats were maintained in 10% +/- 0.5% O2(by displacement with N2) for 3 weeks, whereas in the other, rats were maintained in air. The single smooth muscle cell was isolated from pulmonary artery (phi 200-700 microns) of Wistar rats with acute enzymatic digestion method. Using patch-clamp technique, we recorded the outward K+ currents in pulmonary artery smooth muscle cells and identified a Ca2+.ATP activated K+ channel (K+Ca-ATP) and a delayed rectifier K+ channel among the outward K+ currents. We compared the activities of Ca2+.ATP activated K+ channel (K+Ca-ATP) or delayed rectifier K+ channel in smooth muscle cells isolated from pulmonary artery of chronic hypoxic and normoxic rats. RESULT: The activities of Ca2+.ATP activated K+ channel (K+Ca-ATP) and delayed rectifier K+ channel in chronic hypoxic group are much lower than that in normal group (T test, P < 0.01). Cromakalim (10 mmol) caused a marked enhancement of activity of the reduced K+Ca-ATP but not the delayed rectifier K+ channel in rats of hypoxic group. CONCLUSION: The persistent decrease of potassium channel activity may contribute to setting the development of chronic hypoxic pulmonary hypertension. Cromakalim, one of potassium channel openers, can decrease the pulmonary hypertension induced by chronic hypoxia and may be a new effective drug for treatment of hypoxic pulmonary hypertension. PMID- 11477899 TI - [Studies on pulmonary fibrosis induced by interstitial macrophages]. AB - OBJECTIVE: To evaluate the role of interstitial macrophages (IMs) in modulating the proliferative capacity of fibroblast(FB). METHOD: IMs isolated by collagenase IA digestion of disaggreted lung tissues from bleomycin-treated rats were studied in this regard. The culture supernatants of alveolar macrophages(AMs) and of IMs were added to culture media of FBs. RESULT: The releases of growth factors by the two sub-populations of macrophage were already elevated on 1st day following bleomycin instillation, and peaked on 7th day at a level nearly 3-fold above that of control groups(IM 342%, AM 332%, P < 0.01), the proliferative capacity was correlated well with the levels of TNF alpha and PDGF (r = 0.657, 0.713, P < 0.01). CONCLUSION: Not only AMs, but also IMs could release growth factors to modulate the proliferation of FBs in vitro. IMs might be of more significance in maintenance of the process of pulmonary fibrosis because of its proximity to FBs. PMID- 11477900 TI - [Anti-asthmatic effects of macrolide antibiotics]. PMID- 11477901 TI - [Progress of basic and researches on Alzheimer disease]. PMID- 11477902 TI - [Difficulties in enhancing efficacy of HIV/AIDS and it's breakthrough points]. PMID- 11477903 TI - [Angiographic observation of immediate effect of electric pulse stimulation at Zhiyang point on coronary artery]. AB - OBJECTIVE: To evaluate angiographical diameter and pressure changes of coronary artery and heart rate in patients with coronary heart disease (CHD) or suspected CHD immediately after electric pulse stimulation of Zhiyang (Du. 9) point. METHODS: Twenty-four patients including 21 males and 3 females with an average age of 48.7 years. Parameters of electric pulse stimulation were as follows: the positive pulse range was 440 V, the negative pulse range was 160 V, the cycle was 8 ms, frequency was 125 Hz, the positive pulse width was 0.8 ms, the negative was 0.4 ms, and the output voltage 9 V. Coronary arteriography was performed by using Judkins technique, and intracoronary pressure with heart rate and any response of patients to the stimulation were recorded pre- and post-stimulation for 90 seconds. The relative diameter of left main coronary artery (LMCA), left anterior descending artery (LAD), left circumflex coronary artery (LCx), and right coronary artery (RCA) before and after the stimulation was measured in the same magnification and their results with recordings of intracoronary pressure and heart rate was analyzed. RESULTS: A mild dilation effect was observed in all coronary arteries with an average dilation of 8.3% (6.3%-15%) in LMCA, 7.7% (5% 11.8%) in LAD, 8.5% (6.7%-11.1%) in LCx, and 9.1% (6.1%-13.3%) in RCA. No significant difference was found in the change of the diameter of LMCA, LAD, LCx and RCA before and after the stimulation and between them. No significant change of an intracoronary pressure was shown in three patients, an increase of intracoronary systolic and diastolic pressure by 36% and 13% was identified in one, and a decrease by 21% and 31% in another patient. Heart rate was decreased by 2.3% in those five patients. CONCLUSIONS: Electric pulse stimulation of Zhiyang point could immediately produce mild dilation effect on coronary arteries, slight decrease on the heart rate, and different effects was produced in intracoronary pressure. PMID- 11477904 TI - [Effects in tetramethylpyrazine on TXA2 and PGI2 by cardio-pulmonary bypass in congenital heart diseases with pulmonary hypertension patients]. AB - OBJECTIVE: To study protective effects of tetramethylpyrazine (TMP) on the imbalance of TXA2/PGI2 during cardiopulmomary bypass (CPB) in congenital heart disease (CHD) with pulmonary hypertension (PH) patients. METHODS: Thirty patients suffered from non-cyanotic CHD-PH were randomly divided into control group (n = 15) and treatment group (n = 15). 3 mg/kg of TMP was dripped intravenously after anesthesia and 1 mg/kg of TMP was infused into oxygenator after CPB individually. Blood samples were collected and TXA2 as well as PGI2 were measured after anesthesia induction, 15 min during CPB, 5 min after release of the aortic cross clamp, and 20 min, 6 hrs and 24 hrs after CPB. RESULTS: There was significant difference between treatment group and control group except before operation and 24 hrs after CPB. CONCLUSIONS: The imbalance of TXA2 and PGI2 in patients with CHD-PH during CPB could correct by TMP. PMID- 11477905 TI - [Study on changes of plasma substance P in essential hypertension with left ventricular hypertrophy patients and the effect of promoting blood circulation and eliminating phlegm]. AB - OBJECTIVE: Exploring the relationship between the plasma substance P(SP) and essential hypertension with left ventricular hypertrophy (EH-LVH) and the effect of promoting blood circulation and eliminating phlegm. METHODS: Before and after treatment, the changes of SP in patients wiith EH-LVH were measured by radioimmunoassay. Meanwhile, the changes of plasma renin activity (PRA), angiotension II (Ang II), left ventricular mass index (LVMI) and mean arterial pressure (MAP) were measured. RESULTS: The level of plasma SP in patients with EH LVH was markedly lower than that of patients without EH-NLVH and healthy subjects (P < 0.001); the levels of PRA and Ang II were higher than those in control group (P < 0.01 or < 0.05). After treatment, the level of SP was significantly increased, while LVMI, MAP, PRA and Ang II were significantly decrease (P < 0.01 or < 0.05). The levels between SP and LVMI, MAP, PRA, Ang II were markedly negative correlated (P < 0.01) while levels between LVMI and PRA, Ang II were markedly positive correlated. CONCLUSIONS: The SP may be involved in the pathogenesis of EH-LVH; the mechanism of promoting blood circulation and eliminating phlegm reversing LVH might be related to its increasing the plasma SP. PMID- 11477906 TI - [Relationship between lipometabolic disorder in stroke with TCM syndrome differentiation and typing]. AB - OBJECTIVE: To analyse the relationship between lipometabolic disorder in stroke with TCM Syndrome Differentiation and Typing. METHODS: The serum lipid, lipoprotein and apo-lipoprotein level of TCM treated group (142 cases) and control group (100 cases) were determined. The patients were subdivided into three Type-groups according to TCM Syndrome Differentiation: Phlegm-Dampness Stagnation (PDS), Yin Deficiency with Yang Hyperactivity (YDYH) and Qi Deficiency with Blood Stasis (QDBS) group. RESULTS: The results showed that the level of TC, TG, HDL, LDL, apoA1, apoB and apoA1/apoB were significantly different in PDS and QDBS as compared with control group (P < 0.05, P < 0.01, respectively), while lipometabolic disorder of PDS was most serious in TCM Syndrome Differentiation of stroke. The level of TG, apoA1/apoB were significantly different in YDYH in comparing with control group (P < 0.05). While the level of TC, HDL, LDL and apoB were insignificantly different than those of control group (P > 0.05). CONCLUSIONS: This study showed that lipometabolic disorder of stroke were correlated with Syndrome Differentiation of TCM. Stroke with PDS and QSBS should give removing Phlegm-Dampness, dredging Meridian and removing blood stasis treatment besides routine treatment. PMID- 11477907 TI - [Effect of ligustrazine on nitric oxide contents in cerebrospinal fluid and plasma of patients with cerebral infarction]. AB - OBJECTIVE: To probe into the significance of Nitric Oxide (NO) in ischemic cerebral damage and effect of ligustrazine on it. METHODS: The NO contents in cerebrospinal fluid (CSF) and plasma of 20 controls and 62 patients with arteriosclerotic thrombotic cerebral infarction (ligustrazine group and common treatment group) were determined with Griess method before and after treatment. RESULTS: The NO content in CSF before treatment was higher in severe type, it was higher than that in moderate type, and than that in mild type, again higher than that in control group (all P < 0.05) and was positively correlated with the size of infarction (P < 0.01). There was no significant difference between the plasma NO content of patients and that of control group (P > 0.05), but there was a trend that plasma NO content decreased as the patient's condition worsened. After treatment, the curative effect and plasma NO content of ligustrazine group were both markedly higher than those of common treatment group (P < 0.05). There were no significant differences between CSF NO contents of the two groups, respectively, and that of control group (P > 0.05). CONCLUSIONS: Excessive NO produced in brain tissue and insufficient plasma NO participate the course of ischemic brain damage. Ligustrazine could increase the contents of plasma NO selectively. PMID- 11477908 TI - [Effects of Yunqitang on both esophageal mucosal morphology and esophageal motility in reflux esophagitis patients]. AB - OBJECTIVE: To observe the effects of Yunqitang (YQT) on both esophageal mucosal morphology and esophageal motility in patients with reflux esophagitis (RE). METHODS: According to Syndrome Differentiation of TCM, 42 RE patients were divided into three groups: Disharmony of Liver and Stomach (D) group, Deficiency Cold of Spleen and Stomach (DC) group, Heat Syndrome caused by depression of Liver Qi (H) group. No. I, II, III of YQT were taken respectively for 4 weeks. Before and after treatment scores of typical symptoms were collected, gastroscope and esophageal motility were measured. RESULTS: (1) The symptom remission rate was 81.1%, there were significant differences between the group DC with group D and Group H (P < 0.01). (2) The esophageal mucosal healing rate was 61.9%, the effective rate was 90.5%, and the ineffective rate 9.5%. There weren't significant difference of effective rates among the three groups (P < 0.05). (3) The changes of esophageal motility: lower esophageal sphictor pressure (LESP), average peristaltic pressure (APP) of group D were higher (P < 0.05), LESP, gastro-esophageal barrier pressure (GEBP) and peristaltic conduct speed (PCS) of group DC were remarkably higher (P < 0.05), GEBP of group H was improved (P < 0.05). CONCLUSIONS: YQT has a good therapeutical effect, it's not only resolving reflux symptoms, healing esophageal mucosa, but also improving esophageal motile function. PMID- 11477909 TI - [Treatment of simple obesity in children with photo-acupuncture]. AB - OBJECTIVE: To study the treatment of simple obesity in children and observe its effect. METHODS: Two hundred and two patients were treated with self-produced photo-acupuncture apparatus (101 cases) or ear-pressing (101 cases), and their effects were observed and compared. RESULTS: Effects of both methods were satisfactory. The photo-acupuncture showed a better result than that of the ear pressing. After treatment, the obesity indexes lowered significantly and levels of blood lipids, glucose, cortisol and triiodothyronine were all improved markedly. CONCLUSIONS: Photo-acupuncture is a safe, painless, nontraumatic and effective method for treatment of simple obesity, and it is easy to be accepted by children. PMID- 11477910 TI - [Treatment of 34 cases of chronic aplastic anemia using prepared Rehmannia polysaccharide associated with stanozolol]. AB - OBJECTIVE: To investigate the effect and side effect of prepared Rehmannia polysaccharide (PRP) associated with stanozolol in treating chronic aplastic anemia (CAA). METHODS: Thirty-four cases of CAA were treated by PRP liquid associated with stanozolol, course of 3 week treatment, and control group (17 cases of CAA) was treated by stanozolol alone. RESULTS: Effective rate was 85.3% in treatment group, and that of control group was 58.8%, there was significant difference between them (P < 0.05). Symptom remission of treatment group was earlier than that of control group, symptom scoring between two groups, the difference was highly significant (P < 0.01). Blood cell had a distinct elevation after treatment with PRP associated with stanozolol, and had a significant difference in comparing with pre-treatment in treatment group (P < 0.01). Side effect of PRP was not found during treatment of CAA. CONCLUSIONS: PRP associated with stanozolol to treat CAA has an additional effect, it can accelerate symptom remission without any additional side effect. PMID- 11477911 TI - [Study on 102 cases of anorectal post-operational pain treated by long-acting analgesic]. AB - OBJECTIVE: To observe the clinical effect and experimental result of long-acting analgesic in the treatment of anorectal post-operational pain. METHODS: Analysing the pathological features of anorectal post-operational pain, and select the Radix Stephana Tetrandra, which could disperse pathogenic Wind and dampness, relieve spasm and pain, as the major component. The authors extracted Sinomenine from the Stephana and by mixing with methylene blue and bupivacaine hydrochloride, an injection was prepared. Three groups were in the clinical observation: Long-acting analgesic group (102 cases), routine treatment group (80 cases) and the control group treated by Tramal (30 cases). The curative effect was observed and animal experiment was conducted. RESULTS: The total analgesic rate of the long-acting analgesia was 56.86%. The clinical effect was better than using Bupivacaini hydrochloride or Tramal alone (P < 0.05). It was shown in experiment that long-acting analgesic had obvious effects of topical analgesic and muscle relaxant. CONCLUSIONS: Long-acting analgesic has significant effect in relieving the anorectal post-operational pain. PMID- 11477912 TI - [Experimental study on therapeutical effect of Chinese medicine on polymyositis in guinea pigs]. AB - OBJECTIVE: To study the therapeutical effects of Radix Bupleuri and Ramulus Cinnamomi on virus polymyositis in guinea pigs. METHODS: Forty polymyositis guinea pigs induced by Coxsackievirus B1 were divided at random into A(n = 15), B(n = 15), C(n = 10) groups. A, B groups were treated with Radix Bupleuri and Ramulus Cinnamomi respectively. C group received no treatment. Sign, muscle enzyme spectrum and pathological changes were compared. RESULTS: A and B groups became better. C group showed an aggravating trend (P < 0.05). CONCLUSION: Radix Bupleuri and Ramulus Cinnamomi had therapeutic effect on polymyositis in guinea pigs. PMID- 11477913 TI - [Effects of composite blood-activating decoction on bone marrow microenvironment in mice of immune-induced aplastic anemia]. AB - OBJECTIVE: To explore the mechanism of elevating efficacy for aplastic anemia (AA) by using blood-activating and stasis-eliminating drugs. METHODS: Immune induced aplastic anemia model was established. Each mouse was gastrogavaged by 0.2 ml 100% composite blood-activating decoction (CBAD) twice a day. On the 10th day, the bone marrow histology, CFU-F, adhesive function of the cultured stromal cell layer, bone marrow PO2 were observed. RESULTS: In CBAD group, the WBC count, bone marrow karyocytes, bone marrow hematopoietic tissue volume, CFU-F count were significantly higher than those in AA group (P < 0.01). Moreover, the adhesive function of stromal cells and bone marrow PO2 recovered to normal level. CONCLUSION: The commonly used composite blood-activating decoction could promote the recovery and oxygen-supply of bone marrow microenvironment in AA mice, and improved the bone marrow hematopoiesis. PMID- 11477914 TI - [Effect of electroacupuncture on succinate dehydrogenase of gentamycin induced ototoxic cochlear hair cells in guinea pigs]. AB - OBJECTIVE: To observe the effect of electroacupuncture (EA) on succinate dehydrogenase (SDH) of gentamycin (GE) induced ototoxic cochlear hair cells. METHODS: Preyer's reflex normal guinea pigs were selected and divided randomly into three groups: GE group, EA group, control group. Brainstem auditory evoked potential (BAEP) and SDH in the cochlear hair cells were taken as indexes. In the animals of GE group GE was alone injected intramuscularly 80 mg.kg-1.d-1 for 20 days, while in the animals of EA group GE and additional EA was applied once a day on Tinggong (SI19), Yifeng(SJ17) and Shenshu (UB23) points. EA lasted for 15 minutes. RESULTS: In the GE group BAEP reaction threshold rose markedly, while that rose slightly in EA group. The difference was significant between two groups (P < 0.05). The change of SDH within cochlear hair cells and degree of hair cells injury in the EA group were lower than those in GE group. CONCLUSIONS: EA therapy could relieve GE ototoxicity, protect SDH in cochlear hair cells and might be a possible mechanism of action of EA. PMID- 11477915 TI - [Effect of Fucong tablets and its ingredients on ototoxicity of gentamycin in guinea pigs]. AB - OBJECTIVE: To assess the antagonistic effect of Fucong Tablet (FCT) and its ingredients on ototoxicity of gentamycin in guinea pigs. METHODS: The guinea pigs were given gentamycin 80 mg/kg intramuscularly once daily for 20 days and FCT or its ingredients separately oral taken for 20 days. The auditory brain stem evoked response (ABR) threshold, vascular stria blood circulation condition, outer hair cell count and cochlear ultrastructure of guinea pigs were observed before and after treatment to assess the antagonistic effect and protective effect of FCT on cochlear structure. RESULTS: FCT and its ingredients of tonifying Kidney, replenishing Qi and blood, promoting blood circulation could lower the impairment ratio of outer hair-cell induced by gentamycin (P < 0.05), protect morphosis and function of cochlea and lower the elevated ABR threshold (P < 0.05). CONCLUSIONS: FCT and its ingredients could reduce the toxic damage on ear induced by gentamycin effectively. PMID- 11477916 TI - [Experimental animals in integrated Chinese and western medical research and its prospects]. PMID- 11477917 TI - [Current study of ulcerative colitis treated by rectal administration of Chinese herbal drugs]. PMID- 11477918 TI - [Progress in the treatment of hypertension-induced left ventricular hypertrophy with Chinese traditional medicine]. PMID- 11477919 TI - [Looking forward to the prospect of traditional Chinese medicine in 21st century]. PMID- 11477920 TI - [Effect on life span and membrane protein in red blood cells by integrated medicine therapy on chronic aplastic anemia]. AB - OBJECTIVE: To explore the mechanism of integrated traditional Chinese and Western medicine (TCM-WM) therapy on chronic aplastic anemia (CAA). METHODS: The RBC life span of 30 normal human subjects and 30 patients with CAA were measured by 51Cr labelled technique before and after TCM-WM therapy. The morphology and distribution of RBC membrane protein granules were observed by freeze fracture etching and transmission electron microscope. RESULTS: The half life of erythrocytes (RBC T1/2) was shortened in CAA cases and there was a significant difference compared to healthy control (P < 0.01). After therapy, the RBC life span prolonged and approached the normal level. Before treatment, there existed abnormal in morphology, decrease in amount and uneven in distribution of protein granules in protoplasmic face (PF) and extracellular face (EF) of RBC membrane. After treatment, the protein granules of RBC membrane was improved and approached to control. CONCLUSIONS: The morphology, amount, quality and distribution of RBC membrane protein granule were closely related to its life span. The therapeutic effect of TCM-WM was better than that of WM alone and it had a function both in stabilizing membrane protein and extending the RBC life span. PMID- 11477921 TI - [Study on relativity between oxygen free radical and thromboxane B2, 6-keto-PGF1 alpha during ligustrazine treatment of intrauterine growth retardation]. AB - OBJECTIVE: To study the pathophysiological changes and effective treatment of intrauterine growth retardation (IUGR). METHODS: Eighty-five cases with normal pregnancy and 58 IUGR women maternal red blood cell superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), serum lipid peroxide (LPO) and thromboxane B2 (TXB2), 6-keto-PGF1 alpha(6KP) were examined. IUGR group was divided into Ligustrazine group (LIG, 47 cases) and amino acid group (AAG, 11 cases) and treated accordingly. RESULTS: In IUGR group, LPO increased and SOD, GSH-Px activity, 6KP value decreased abnormally, TXB2/6KP ratio significantly increased, which was negative correlated with SOD, GSH-Px activity and positive correlated with LPO level markedly. After treatment, all the changes were improved and almost reached normal range. The fetal outcome showed that the total effective rate was 95.7% in LIG, markedly higher than that in AAG (81.8%, P < 0.05), there was significant difference between LIG and AAG. CONCLUSIONS: The balance of body oxidate/antioxidate system was disturbed and the uteroplacental-fetal circulation was obstracted in IUGR. Ligustrazine could inhibit oxygen free radical, rise SOD, GSH-Px activity, regulate TXA2/PGI2 balance, promote fetal growth. PMID- 11477922 TI - [Clinical study on Zishen Decoction in chronic uric acid nephrosis]. AB - OBJECTIVE: To observe the ameliorative effect of Zishen Decoction (ZSD) on chronic uric acid nephrosis (CUAN). METHODS: The 72 CUAN patients were divided into 2 groups: The group with ZSD treatment, the dose of which was 400 ml/d and group with zyloric as the control, the dose of which was 200 mg/d. The treatment lasted for 8 weeks. RESULTS: The general effective rate of the ZSD group was 92.86%; and that of the control group Was 66.67%. There was significant difference between the two groups (P < 0.01). ZSD treatment reduced the levels of blood uric acid, serum creatinine and blood urea nitrogen and the levels of albumin, beta 2-microglobulin and the activity of N-acetyl-D-glucosaminidase in CUAN (P < 0.01). The levels of triglyceride and total cholesterol decreased and the levels of high density lipoprotein cholesterol increased in the serum of ZSD treated CUAN (P < 0.01). CONCLUSIONS: ZSD exerted obviously ameliorative effect on renal function in CUAN. PMID- 11477923 TI - [Clinical study on preventing and treating chemotherapy induced nausea and vomiting using supplemented Inula-Ochrae Decoction]. AB - OBJECTIVE: To observe supplemented Inula-Ochrae Decoction (SIOD) in preventing and treating nausea and vomiting induced by chemotherapy for patients with malignant tumour. METHODS: Seventy-two patients were divided into two groups, the patients in test group took SIOD and in control group using ondensetron, and the efficacy of SIOD in preventing and treating chemotherapy induced nausea and vomiting were studied prospectively with self-intersection approach. RESULTS: The effective rate of treating vomiting due to chemotherapy with DDP and without DDP in the test group was 92.7% and 93.5% respectively, and was higher than that in the control group 87.8% and 87.1%, the difference was insignificant statistically (P > 0.05). CONCLUSIONS: The prescription SIOD could prevent and treat effectively chemotherapy induced nausea and vomiting without any toxic and side effects, and is inexpensive with high efficacy for clinical use. PMID- 11477924 TI - [Clinical study on effect of Fuzheng Kangbai Granule on long-term survival of patients with acute leukemia]. AB - OBJECTIVE: To observe the effect of Fuzheng Kangbai Granule (FZKBG) on event free interval (EFI) and over survival (OS) of patients with acute leukemia, and to study the mechanism of FZKBG. METHODS: FZKBG was used in 90 cases of completely remitted acute leukemia, immune functions of patients before and after using FZKBG were measured. RESULTS: Five year EFI and OS were 64.2% and 77.2% of 90 cases of completely remitted acute leukemia, and the immune functions after using FZKBG have improved significantly. CONCLUSIONS: FZKBG could increase the EFI and OS of patients with acute leukemia, and the improved immune functions may play a role in increasing 5 year EFI and OS. PMID- 11477925 TI - [Augmented effect of Composite Radix Salviae Miltiorrhizae Injection on radiosensitivity of leukemia cells]. AB - OBJECTIVE: To explore the effect of Composite Radix Salviae Miltiorrhizae Injection (CSMI) on radiosensitivity of leukemia cells. METHODS: Semisolid agar culture and flow cytometry assay were performed for studying the change in radiosensitivity of HL60 cell line and fresh human leukemia cells after exposing to CSMI. RESULTS: The D0(the inverse of the slope of the survival curve) and SF2(survival fraction at 2 Gy) of HL60 cell line were decreased from 1.53, 0.34 to 0.93, 0.12, respectively and apoptosis rates after radiation were raised significantly by CSMI. Furthermore, the concentration of CSMI and the time of mixed culture with CSMI before irradiation had positive relation to the effects mentioned above. Compared with control group, CSMI could increase the radiation induced apoptosis of fresh leukemia cells (5.89 +/- 2.91% vs 12.05 +/- 3.06%). CONCLUSION: CSMI could obviously enhance the radiosensitivity of leukemia cells. PMID- 11477926 TI - [Clinical study of puerarin in treatment of patients with unstable angina]. AB - OBJECTIVE: To observe the clinical efficacy of puerarin in treating patients with unstable angina (UA). METHODS: Frequency of angina events, consumed doses of nitrates, heart rate, blood pressure, rest electrocardiogram (ECG) and myocardial oxygen consumption were observed before and after drug therapy in puerarin group (21 cases) and control group (18 cases) respectively. RESULTS: The puerarin group not only showed more effective than that in control group in reducing frequency of angina events and consumed doses of nitrates and improving abnormal rest ECG, but also appeared to be preferable to the control group in reducing myocardial oxygen consumption and increasing exercise duration (P < 0.05). CONCLUSION: Puerarin is a safe and effective drug in treating patients with UA. PMID- 11477927 TI - [Guiding-evaluation of immunomodulating activity of the Liuwei Dihuang Decoction during the stepwise fractionation]. AB - OBJECTIVE: To conduct the activity evaluation for immunomodulating components from the Liuwei Dihuang Decoction (LWDHD). METHODS: Cyclophosphamide (Cy)-treated mice were used as the immunodeficient model and the antibody production response in plaque-forming cell (PFC) assay was employed as the activity-evaluating parameter. Stepwise fractionation was guided by activity-evaluation. RESULTS: Fr1, the ethanol-soluble fraction, and Fr2, the ethanol insoluble fraction of LWDHD, both significantly improved the antibody production response in Cy-treated mice, in which Fr2 showed stronger activity than that of Fr1. Fr2 was further fractionated. Fr2-CA4, one of the fractions obtained by active carbon chromatography from Fr2, exhibited the strongest activity in comparison with other fractions. CONCLUSION: CA4, which is mainly composed of acidic polysaccharides, is the main immunomodulating active fraction contained in the ethanol-insoluble fraction of LWDHD. PMID- 11477928 TI - [Study on efficacy of Chinese Kidney-Tonifying Recipe in male rats with osteoporosis induced by dexamethasone and its mechanism]. AB - OBJECTIVE: To study the efficacy of traditional Chinese Kidney-Tonifying Recipe (KTR) in male rats with osteoporosis induced by dexamethasone (DM) and its mechanism. METHODS: Forty-six male Sprague-Dawley rats at 12 months of age were chosen: they were divided into four groups and KTR and DM were administered appropriately for 18 weeks. General bone mineral density, femur bending strength, serum-BGP, serum-PTH and serum-E2 were measured. RESULTS: General bone mineral density, femur bending strength in KTR prevention group were significantly higher than those in DM group (P < 0.01) and the same with those in normal control group (P > 0.05). General bone mineral density, femur bending strength in KTR treatment group were higher than those in DM group (P < 0.05), and lower than those in normal control group. Compared with DM group, level of serum-T in KTR prevention group and KTR treatment group increased (P < 0.05) and level of serum-PTH in those two groups decreased (P < 0.05). But in comparison with DM group, level of serum-E2 and serum-BGP in the above-mentioned two groups was unchanged (P > 0.05). CONCLUSIONS: The data indicated that traditional Chinese Kidney-Tonifying Recipe could prevent and treat osteoporosis in male rats induced by DM. PMID- 11477929 TI - [Effect of water extract of Poria on cytosolic free calcium concentration in brain nerve cells of neonatal rats]. AB - OBJECTIVE: To investigate the effect of Poria cocos, a Chinese medicinal herb, on cytosolic free calcium concentration ([Ca2+]i) in brain nerve cells of neonatal rats. METHODS: Double-wave-length fluorospectrophotometer and laser confocal scanning microscope were used to measure the [Ca2+]i in neurons. RESULTS: The water extract of poria (31-250 mg/L) induced a dose-dependent increase in [Ca2+]i, but poria at the range of 500 mg/L did not cause [Ca2+]i to increase. It was also shown that the water extract of poria significantly inhibited the increment of [Ca2+]i induced by 500 mumol/L glutamate. The similar results were observed from the experiments in primarily cultured hippcampal neurons of neonatial rats by using laser confocal scanning microscope. CONCLUSIONS: The water extract of Poria cocos has the effect of bidirectional regulation on cytosolic free calcium in brain nerve cells. PMID- 11477930 TI - [Effect of Zhongfeng Naodeping Granule on hippocampal excitatory amino acid and neuron of stroke-prone spontaneously hypertensive rats with hemorrhagic apoplexy]. AB - OBJECTIVE: To observe the effects of Zhongfeng Naodeping Granule (ZFNDPG) on hemorrhagic apoplexy. METHODS: The stroke-prone spontaneously hypertensive rats (SHRsp) were used to study effects of ZFNDPG on hemorrhage apoplexy. Excitatory amino acid (EAA) concentration in hippocampus sector, neuronal density and ultrastructural changes in hippocampal CA1 sector were measured. RESULTS: In pathological model group glutamate (Glu) and aspartate (Asp) concentration elevated obviously. With the ZFNDPG treating SHRsp of hemorrhagic apoplexy, Glu and Asp concentration in hippocampal sector could be markedly inhibited, compared with model group, P < 0.05-0.01. Neuronal morphology was observed: neurone injury was mild and neuronal density increased in hippocampal CA1 sector of treatment group, compared with model group, P < 0.01. Electron microscopy showed: edema, degeneration and necrosis caused by hemorrhagic apoplexy were improved after the ZFNDPG treatment. CONCLUSIONS: Effects of protecting neurone for SHRsp on hemorrhagic apoplexy might be associated with that ZFNDPG inhibited concentration of EAA. PMID- 11477931 TI - [Exploration on therapeutic mechanism of warm-hot herbal drugs]. PMID- 11477932 TI - [Progress in the study of antineoplastic activity of suphora flarescens ait and its alkaloids]. PMID- 11477933 TI - Biological markers of long-term effects of naturally occurring stress. PMID- 11477934 TI - Behavior: the endocrine-immune interface and health outcomes. PMID- 11477935 TI - Hypertension at the workplace--an occult disease? The need for work site surveillance. PMID- 11477936 TI - Psychosocial stress and the physiology of atherosclerosis. PMID- 11477937 TI - Regulatory disturbance of energy. PMID- 11477938 TI - Endocrinological assessment of extreme stress. PMID- 11477939 TI - Salivary cortisol response to stress in children. PMID- 11477940 TI - Multiple physiological assessments of long-term stress at work and in daily life: a system approach. PMID- 11477941 TI - Finding a stress measure in the literature and taking it into the field. PMID- 11477942 TI - Epidemiological applications of long-term stress in daily life. PMID- 11477943 TI - Long-term stress in daily life in a socioepidemiologic perspective. PMID- 11477944 TI - Forget HIPAA; think of your patients. PMID- 11477945 TI - Priorities and strategies. PMID- 11477946 TI - How secure is your palm-top? PMID- 11477947 TI - Four tips for EMR integration. PMID- 11477948 TI - When all else fails, contact your HCFA regional office. PMID- 11477949 TI - Back to school: options for learning the business of medicine. PMID- 11477950 TI - Depositions: defending your care. PMID- 11477951 TI - A problem-oriented approach to the HIPAA security standards. PMID- 11477952 TI - Responding to an unfavorable quality assurance audit. PMID- 11477953 TI - Buying your first PDA. PMID- 11477954 TI - Protecting the time you've got. PMID- 11477955 TI - A challenge to the American Medical Association. PMID- 11477956 TI - Risk adjustment for measuring health outcomes: an application in VA long-term care. AB - An empirically derived risk adjustment model is useful in distinguishing among facilities in their quality of care. We used Veterans Affairs (VA) administrative databases to develop and validate a risk adjustment model to predict decline in functional status, an important outcome measure in long-term care, among patients residing in VA long-term care facilities. This model was used to compare facilities on adjusted and unadjusted rates of decline. Predictors of decline included age, time between assessments, baseline functional status, terminal illness, pressure ulcers, pulmonary disease, cancer, arthritis, congestive heart failure, substance-related disorders, and various neurologic disorders. The model performed well in the development and validation databases (c statistics, 0.70 and 0.68, respectively). Risk-adjusted rates and rankings of facilities differed from unadjusted ratings. We conclude that judgments of facility performance depend on whether risk-adjusted or unadjusted decline rates are used. Valid risk adjustment models are therefore necessary when comparing facilities on outcomes. PMID- 11477957 TI - Organizational culture, continuous quality improvement, and medication administration error reporting. AB - This study explores the relationships among measures of nurses' perceptions of organizational culture, continuous quality improvement (CQI) implementation, and medication administration error (MAE) reporting. Hospital-based nurses were surveyed using measures of organizational culture and CQI implementation. These data were combined with previously collected data on perceptions of MAE reporting. A group-oriented culture had a significant positive correlation with CQI implementation, whereas hierarchical and rational culture types were negatively correlated with CQI implementation. Higher barriers to reporting MAE were associated with lower perceived reporting rates. A group-oriented culture and a greater extent of CQI implementation were positively (but not significantly) associated with the estimated overall percentage of MAEs reported. We conclude that health care organizations have implemented CQI programs, yet barriers remain relative to MAE reporting. There is a need to assess the reliability, validity, and completeness of key quality assessment and risk management data. PMID- 11477958 TI - Use of screening and preventive services among women with disabilities. AB - Roughly 54 million Americans have some disability; at older ages, women are more likely to be disabled than men. Many people with disabilities today live virtually normal life spans, and therefore routine screening and preventive services are essential to their overall quality of care. We used the 1994-1995 National Health Interview Survey (NHIS), with Disability, Family Resources, and Healthy People 2000 supplements, to examine screening and preventive service use for adult women with disabilities living in the community--about 18.4% of women (estimated 18.28 million). Disability was associated with higher age-adjusted rates of: poverty; living alone; low education; inability to work; obesity; and being frequently depressed or anxious. Disabled women generally reported screening and preventive services at rates comparable to all women. Women with major lower extremity mobility difficulties had much lower adjusted odds of Papanicolaou smears (odds ratio, 0.6; 95% confidence interval, 0.4-0.9), mammograms (odds ratio, 0.7; 95% confidence interval, 0.5-0.9), and smoking queries (odds ratio, 0.6; 95% confidence interval, 0.5-0.8). Various approaches exist to improve access for disabled women to health care services. PMID- 11477959 TI - Seroprevalence of hepatitis A among hospital dietary workers: implications for screening and immunization. AB - Hepatitis A is a self-limited, virally mediated infection of the liver. The usual mode of transmission is by the fecal-oral route. Employees of food-service establishments who are infected with the hepititus A virus can transmit the disease when handling food products. The Centers for Disease Control and Prevention recommends the use of the hepatitis A vaccine among dietary workers who may be at risk for contracting and spreading the disease. Because hepatitis A infection can often be a subclinical disease, the incidence of cases reported is not indicative of its true prevalence. The objective of this study was to document the seroprevalence of hepatitis A among hospital dietary workers. Dietary workers at a suburban hospital were interviewed to determine if they had been exposed to hepatitis A and if they had a history of hepatitis A infection. Serum was obtained from each subject and tested for the presence of hepatitis A antibodies. The Abbott HAVAB EIA kit was used for the detection of immunoglobulin G (IgG) and IgM hepatitis A antibodies. Of 119 subjects, 56 (47%) were women, and 63 (53%) were men; the subjects had a mean age of 42 years (range, 20-66). Fifty one subjects (43%) were born in the United States, and 68 subjects (57%) were born outside of the United States. Of the 119 subjects, only 2 (2%) had a known history of hepatitis A infection, yet 67 (56%) had hepatitis A titers; 52 (44%) were susceptible to the disease. One subject had received the hepatitis A vaccine. Fifty-five of 68 foreign-born subjects (81%; 95% confidence interval, 71 91%) had hepatitis A antibodies versus 12 of 51 US-born subjects (24%; 95% confidence interval, 12-36%). The foreign-born subjects had a mean age of 28 years at the time of their arrival in the United States. In conclusion, a large number of foreign-born hospital dietary workers have hepatitis A antibodies without a history of disease. Immunization of this group of dietary workers may not have any beneficial effects, nor is it cost-effective. PMID- 11477960 TI - Risk-pooling--necessary but not sufficient? PMID- 11477961 TI - Has oral fluid the potential to replace serum for the evaluation of population immunity levels? A study of measles, rubella and hepatitis B in rural Ethiopia. AB - OBJECTIVE: To assess the suitability of using oral-fluid samples for determining the prevalence of immunity to vaccine-preventable infections. METHODS: Paired blood and oral-fluid samples were obtained from 853 individuals of all ages from a rural Ethiopian community. Oral fluid around the gums was screened for measles- and rubella-specific antibodies using enhanced IgG antibody capture (GAC) enzyme linked immunosorbent assays (ELISAs), and for anti-HBc antibodies using a prototype GACELISA. IgG antibodies in serum to measles, rubella and HBc were determined using commercial ELISAs. FINDINGS: Relative to serum, oral fluid assay sensitivity and specificity were as follows: 98% and 87% for measles, 79% and 90% for rubella, and 43% and 87% for anti-HBc. These assay characteristics yielded population prevalence estimates from oral fluid with a precision equal to that of serum for measles (all ages) and rubella (ages < 20 years). CONCLUSION: Our results suggest that oral fluid could have the potential to replace serum in IgG antibody prevalence surveys. Further progress requires assessment of variation in assay performance between populations as well as the availability of standardized, easy to use assays. PMID- 11477962 TI - Modelling HIV/AIDS epidemics in sub-Saharan Africa using seroprevalence data from antenatal clinics. AB - OBJECTIVE: To improve the methodological basis for modelling the HIV/AIDS epidemics in adults in sub-Saharan Africa, with examples from Botswana, Central African Republic, Ethiopia, and Zimbabwe. Understanding the magnitude and trajectory of the HIV/AIDS epidemic is essential for planning and evaluating control strategies. METHODS: Previous mathematical models were developed to estimate epidemic trends based on sentinel surveillance data from pregnant women. In this project, we have extended these models in order to take full advantage of the available data. We developed a maximum likelihood approach for the estimation of model parameters and used numerical simulation methods to compute uncertainty intervals around the estimates. FINDINGS: In the four countries analysed, there were an estimated half a million new adult HIV infections in 1999 (range: 260 to 960 thousand), 4.7 million prevalent infections (range: 3.0 to 6.6 million), and 370 thousand adult deaths from AIDS (range: 266 to 492 thousand). CONCLUSION: While this project addresses some of the limitations of previous modelling efforts, an important research agenda remains, including the need to clarify the relationship between sentinel data from pregnant women and the epidemiology of HIV and AIDS in the general population. PMID- 11477963 TI - Neonatal mortality of low-birth-weight infants in Bangladesh. AB - OBJECTIVE: To ascertain the role of low birth weight (LBW) in neonatal mortality in a periurban setting in Bangladesh. METHODS: LBW neonates were recruited prospectively and followed up at one month of age. The cohort of neonates were recruited after delivery in a hospital in Dhaka, Bangladesh, and 776 were successfully followed up either at home or, in the event of early death, in hospital. FINDINGS: The neonatal mortality rate (NMR) for these infants was 133 per 1000 live births (95% confidence interval: 110-159). The corresponding NMRs (and confidence intervals) for early and late neonates were 112 (91-136) and 21 (12-33) per thousand live births, respectively. The NMR for infants born after fewer than 32 weeks of gestation was 769 (563-910); and was 780 (640-885) for infants whose birth weights were under 1500 g. Eighty-four per cent of neonatal deaths occurred in the first seven days; half within 48 hours. Preterm delivery was implicated in three-quarters of neonatal deaths, but was associated with only one-third of LBW neonates. CONCLUSION: Policy-relevant findings were: that LBW approximately doubles the NMR in a periurban setting in Bangladesh; that neonatal mortality tends to occur early; and that preterm delivery is the most important contributor to the NMR. The group of infants most likely to benefit from improvements in low-cost essential care for the newborn accounted for almost 61% of neonatal mortalities in the cohort. PMID- 11477964 TI - Predicting and comparing long-term measles antibody profiles of different immunization policies. AB - OBJECTIVE: Measles outbreaks are infrequent and localized in areas with high coverage of measles vaccine. The need is to assess long-term effectiveness of coverage. Since 1991, no measles epidemic affecting the whole island has occurred in Taiwan, China. Epidemiological models are developed to predict the long-term measles antibody profiles and compare the merits of different immunization policies on the island. METHODS: The current measles immunization policy in Taiwan, China, is 1 dose of measles vaccine at 9 months of age and 1 dose of measles, mumps and rubella (MMR) vaccine at 15 months of age, plus a 'mop-up' of MMR-unvaccinated schoolchildren at 6 years of age. Refinements involve a change to a two-dose strategy. Five scenarios based on different vaccination strategies are compared. The models are analysed using Microsoft Excel. FINDINGS: First, making the assumption that measles vaccine-induced immunity will not wane, the predicted measles IgG seroprevalences in preschool children range from 81% (lower bound) to 94% (upper bound) and in schoolchildren reach 97-98% in all strategy scenarios. Results are dependent on the association of vaccine coverage between the first and second dose of vaccine. Second, if it is assumed that vaccine induced antibody titres decay, the long-term measles seroprevalence will depend on the initial titres post vaccination, decay rates of antibody titres and cut off of seropositivity. CONCLUSION: If MMR coverage at 12 months of age can reach > 90%, it would be worth changing the current policy to 2 doses at 12 months and 6 years of age to induce higher antibody titres. These epidemiological models could be applied wherever a similar stage of measles elimination has been reached. PMID- 11477965 TI - Efficacy trial of Vi polysaccharide vaccine against typhoid fever in south western China. AB - OBJECTIVE: To test the efficacy of locally produced Vi vaccine over a time period of longer than one year. METHODS: A double-blinded, randomized field trial was performed in Guangxi Zhuang Autonomous Region in south-western China, using 30 micrograms doses of locally produced Vi. Enrolled subjects were 3-50 years of age, although the majority (92%) were school-aged children, who have the highest rate of typhoid fever in this setting. A total of 131,271 people were systematically allocated a single dose of 30 micrograms of Vi polysaccharide or saline placebo. The study population was followed for 19 months, with passive surveillance conducted in the Ministry of Health and the Regional Health and Anti epidemic Centre (HAEC). Clinically suspected cases of typhoid fever were confirmed by blood culture, or by serological reaction with O-antigen (Widal tests). FINDINGS: After 19 months, there were 23 culture-confirmed cases of typhoid fever in the placebo group versus 7 cases in the Vi group (Protective efficacy (PE) = 69%; 95% CI = 28%, 87%). Most of the isolates were from school aged children: 22 cases in the placebo group versus 6 in the Vi group (PE = 72%; 95% CI = 32%, 82%). No serious post-injection reactions were observed. The locally produced Vi polysaccharide vaccine showed levels of protective efficacy similar to those for Vi vaccine produced in industrial countries. CONCLUSION: The slightly higher dose of vaccine did not seem to alter efficacy significantly in China. PMID- 11477966 TI - Efficacy of oral azithromycin versus topical tetracycline in mass treatment of endemic trachoma. AB - OBJECTIVE: To compare the impact of mass treatment with oral azithromycin and topical tetracycline on the prevalence of active trachoma. METHODS: A total of 1803 inhabitants from 106 households of eight Gambian villages were randomized, in pairs, to receive either three doses of azithromycin at weekly intervals, or daily topical tetracycline over 6 weeks. Ocular examinations were conducted before treatment, and 2, 6 and 12 months after treatment. FINDINGS: Prior to treatment, 16% of the study participants had active trachoma. Two months after treatment, the prevalence of trachoma was 4.6% and 5.1% in the azithromycin and the tetracycline groups, respectively (adjusted odds ratio (OR) = 1.09; 95% confidence interval (CI) = 0.53, 2.02). Subsequently, the prevalence rose to 16% in the tetracycline group, while remaining at 7.7% in the azithromycin group (adjusted OR at 12 months = 0.52; 95% CI = 0.34, 0.80). At 12 months post treatment, there were fewer new prevalent cases in the azithromycin group, and trachoma resolution was significantly better for this group (adjusted OR = 2.02; 95% CI = 1.42, 3.50). CONCLUSION: Oral azithromycin therefore appears to offer a means for controlling blinding trachoma. It is easy to administer and higher coverages may be possible than have been achieved hitherto. PMID- 11477967 TI - Operational issues in preventing mother-to-child transmission of HIV-1 in Abidjan, Cote d'Ivoire, 1998-99. AB - OBJECTIVE: To demonstrate the feasibility, from the public health standpoint, of preventing mother-to-child transmission of human immunodeficiency virus type 1 (HIV-1) in Africa. METHODS: Voluntary counselling and HIV serotesting were routinely provided in four health centres in Abidjan, Cote d'Ivoire, for six months in 1998-99. Peripartum treatment with zidovudine and alternatives to breastfeeding were provided free to HIV-infected women. FINDINGS: Of the 4309 pregnant women in the study who attended their first antenatal care visit, 3756 benefited from individual counselling and pretesting (87.2%), and 3452 (80.1%) agreed to undergo HIV serotesting. Overall HIV prevalence was (12.89%) and 5% for women aged under 18 years. Among the 2998 HIV-negative women, 71% returned for their test result, whereas only 60% of the 445 HIV-positive women did so. A total of 124 HIV-positive women were informed of their serostatus and the possibility of preventing mother-to-child transmission of HIV; 100 started treatment and 80 completed zidovudine prophylaxis. At 6 weeks of age, 36 of the 78 liveborn children were being breastfed (46%), two were being mixed-fed and 41 (52%) were being artificially fed. CONCLUSIONS: In Abidjan, voluntary counselling and HIV testing with a view to preventing mother-to-child transmission was feasible in antenatal care units and was well accepted by pregnant women. An insufficient proportion of women returned to obtain their test results. This was especially so among HIV-positive women, the target group for preventing mother-to-child transmission of HIV. Additional staff were required in order to offer voluntary counselling and HIV testing to the study women. Close supervision and strong commitment of health workers were essential. Alternatives to breastfeeding were effectively proposed to HIV-positive women, with active follow-up of children and clinical, nutritional and social support. PMID- 11477968 TI - Three case definitions of malaria and their effect on diagnosis, treatment and surveillance in Cox's Bazar district, Bangladesh. AB - In countries where malaria is endemic, routine blood slide examinations remain the major source of data for the public health surveillance system. This approach has become inadequate, however, as the public health emphasis has changed from surveillance of laboratory-confirmed malaria infections to the early detection and treatment of the disease. As a result, it has been advocated that the information collected about malaria be changed radically and should include the monitoring of morbidity and mortality, clinical practice and quality of care. To improve the early diagnosis and prompt treatment (EDPT) of malaria patients, three malaria case definitions (MCDs) were developed, with treatment and reporting guidelines, and used in all static health facilities of Cox's Bazar district, Bangladesh (population 1.5 million). The three MCDs were: uncomplicated malaria (UM); treatment failure malaria (TFM); and severe malaria (SM). The number of malaria deaths was also reported. This paper reviews the rationale and need for MCDs in malaria control programmes and presents an analysis of the integrated surveillance information collected during the three-year period, 1995 97. The combined analysis of slide-based and clinical data and their related indicators shows that blood slide analysis is no longer used to document fever episodes but to support EDPT, with priority given to SM and TFM patients. Data indicate a decrease in the overall positive predictive value of the three MCDs as malaria prevalence decreases. Hence the data quantify the extent to which the mainly clinical diagnosis of UM leads to over-diagnosis and over-treatment in changing epidemiological conditions. Also the new surveillance data show: a halving in the case fatality rate among SM cases (from 6% to 3.1%) attributable to improved quality of care, and a stable proportion of TFM cases (around 7%) against a defined population denominator. Changes implemented in the EDPT of malaria patients and in the surveillance system were based on existing staff capacity and routine reporting structures. PMID- 11477969 TI - Every death counts: measurement of maternal mortality via a census. AB - Methods for measuring maternal mortality at national and subnational levels in the developing world lag far behind the demand for estimates. We evaluated use of the national population census as a means of measuring maternal mortality by assessing data from five countries (Benin, Islamic Republic of Iran, Lao People's Democratic Republic, Madagascar, and Zimbabwe) which identified maternal deaths in their censuses. Standard demographic methods were used to evaluate the completeness of reporting of adult female deaths and births in the year prior to the census. The results from these exercises were used to adjust the data. In four countries, the numbers of adult female deaths needed to be increased and three countries required upward adjustment of the numbers of recent births. The number of maternal deaths was increased by the same factor as that used for adult female deaths on the assumption that the proportion of adult female deaths due to maternal causes was correct. Age patterns of the various maternal mortality indicators were plausible and consistent with external sources of data for other populations. Our data suggest that under favourable conditions a national census is a feasible and promising approach for the measurement of maternal mortality. Moreover, use of the census circumvents several of the weaknesses of methods currently in use. However, it should also be noted that careful evaluation of the data and adjustment, if necessary, are essential. The public health community is urged to encourage governments to learn from the experience of these five countries and to place maternal mortality estimation in the hands of statistical agencies. PMID- 11477970 TI - Effect of removing user fees on attendance for curative and preventive primary health care services in rural South Africa. AB - User fees are used to recover costs and discourage unnecessary attendance at primary care clinics in many developing countries. In South Africa, user fees for children aged under 6 years and pregnant women were removed in 1994, and in 1997 all user fees at all primary health care clinics were abolished. The intention of these policy changes was to improve access to health services for previously disadvantaged communities. We investigated the impact of these changes on clinic attendance patterns in Hlabisa health district. Average quarterly new registrations and total attendances for preventive services (antenatal care, immunization, growth monitoring) and curative services (treatment of ailments) at a mobile primary health care unit were studied from 1992 to 1998. Regression analysis was undertaken to assess whether trends were statistically significant. There was a sustained increase in new registrations (P = 0.0001) and total attendances (P = 0.0001) for curative services, and a fall in new registrations (P = 0.01) and total attendances for immunization and growth monitoring (P = 0.0002) over the study period. The upturn in demand for curative services started at the time of the first policy change. The decreases in antenatal registrations (P = 0.07) and attendances (P = 0.09) were not statistically significant. The number of new registrations for immunization and growth monitoring increased following the first policy change but declined thereafter. We found no evidence that the second policy change influenced underlying trends. The removal of user fees improved access to curative services but this may have happened at the expense of some preventive services. Governments should remain vigilant about the effects of new health policies in order to ensure that objectives are being met. PMID- 11477971 TI - Reinsurance of health insurance for the informal sector. AB - Deficient financing of health services in low-income countries and the absence of universal insurance coverage leaves most of the informal sector in medical indigence, because people cannot assume the financial consequences of illness. The role of communities in solving this problem has been recognized, and many initiatives are under way. However, community financing is rarely structured as health insurance. Communities that pool risks (or offer insurance) have been described as micro-insurance units. The sources of their financial instability and the options for stabilization are explained. Field data from Uganda and the Philippines, as well as simulated situations, are used to examine the arguments. The article focuses on risk transfer from micro-insurance units to reinsurance. The main insight of the study is that when the financial results of micro insurance units can be estimated, they can enter reinsurance treaties and be stabilized from the first year. The second insight is that the reinsurance pool may require several years of operation before reaching cost neutrality. PMID- 11477972 TI - Setting health priorities: the development of cost-effectiveness league tables. PMID- 11477973 TI - Resource allocation in decentralized health systems. PMID- 11477974 TI - Diethylene glycol poisoning in Gurgaon, Haryana, India, 1998. PMID- 11477975 TI - Treated nets vs house spraying. PMID- 11477976 TI - Europe gets tough on smoking. PMID- 11477977 TI - Countries split over tobacco treaty. PMID- 11477978 TI - Color memory and evaluations for alphabetic and logographic brand names. AB - Reading logographs relies to a greater degree on visual information processes than does reading alphabetic words. Two experiments examined related memory and judgment effects with native speakers of Chinese and English. In Experiment 1, memory for print colors was greater for logographs than for alphabetic words. Experiment 2 examined consumers' ratings of novel brand names printed in colors previously associated with positive or negative evaluations. These print colors had a stronger effect on the evaluation of logographic than of alphabetic brand names. The findings suggest that script variations affect the integration of written words with their surface features. The findings have practical implications for visually differentiating a brand and for the ability of a brand extension or a copycat brand to visually acquire meaning from an existing brand. PMID- 11477979 TI - Color coding information: assessing alternative coding systems using independent brightness and hue dimensions. AB - Can independent dimensions of brightness and hue be used in a combined digital information code? This issue was addressed by developing 2 color-coding systems and testing them on informed and naive participants in signal beam detection and classification experiments for simulated sonar displays. Each coding system's results showed both groups efficiently used encoded information that varied simultaneously along the 2 dimensions of brightness and hue. Findings support the proposed procedures for developing color information codes and the validity of such information codes across different populations. Applied significance of these results is provided by the test of principled methods of color-code construction and the demonstration that extending the information content of user interfaces beyond 1 dimension is feasible in practice. PMID- 11477980 TI - Retention and transfer of morse code reception skill by novices: part-whole training. AB - The training of composite skills requiring differential responding to a large set of stimuli raises issues about how to break down the whole task into parts and which parts should be trained first. Components of Morse code reception skill were identified, separated, and used to test whether initial training on a difficult part was more effective than initial training on an easy part. Initial training on a difficult subset of stimuli and on a difficult subtask both yielded disadvantages rather than the advantage implied by recent findings with different tasks. Incremental training should begin with the part yielding the most effective strategic skills, which appear to depend on characteristics of the task. In both present experiments, easy initial training led to adoption of an effective unitization strategy for representing codes. The hypothesis that procedural reinstatement at delayed testing leads to better retention was supported and extended. PMID- 11477981 TI - Unequal weighting of monocular inputs in binocular combination: implications for the compression of stereoscopic imagery. AB - For efficient storage and transmission of stereoscopic images over bandwidth limited channels, compression can be achieved by degrading 1 monocular input of a stereo pair and maintaining the other at the desired quality. The desired quality of the fused stereoscopic image can be achieved, provided that binocular vision assigns greater weight to the nondegraded input. A psychophysical matching procedure was used to determine if such over-weighting occurred when the monocular degradation included blur or blocking artifacts. Over-weighting of the nondegraded input occurred for blur, but under-weighting of the nondegraded input occurred for blockiness. Some participants exhibited ocular dominance, but this did not affect the blur results. The authors conclude that blur, but not blockiness, is an acceptable form of monocular degradation. PMID- 11477982 TI - Making choices in anticipation of similar future choices can increase self control. AB - Two experiments tested the efficacy of linking a current choice with similar future choices as a means of increasing self-control. Participants were offered choices between smaller and sooner vs. larger and later amounts of money (Experiment 1, n = 60) or food (Experiment 2, n = 34). After a small-large pair for which the participant preferred the smaller reward was found, a choice between the same pair was offered as the 1st of 5 such choices to be offered over a period of weeks. The majority of participants in both experiments who chose between all 5 smaller and all 5 larger rewards chose the larger rewards. One third of participants in Experiment 1 who could choose independently on each pair in the series reversed their previous preference and chose the larger reward in the context of the series. These results suggest that self-control can be enhanced by viewing one's current choice as predictive of future choices. PMID- 11477983 TI - Biased interpretation of evidence by mock jurors. AB - Predecisional distortion is jurors' biased interpretation of new evidence to support whichever verdict is tentatively favored as a trial progresses. In 2 experiments, students and prospective jurors distorted evidence from a mock trial. Further, the magnitude of prospective jurors' distortion was twice that of students. Consistent with previous research, distortion increased with juror confidence in whichever verdict was currently leading. In spite of clear instructions to ignore prior beliefs, general proplaintiff or prodefendant attitudes influenced the verdicts of prospective jurors, but not of students. These findings suggest that jury instructions should warn against not only premature decisions but also any tentative judgments, lest such opinions influence jurors' evaluations of subsequent evidence. Predecisional distortion in jury trials may lead to biased outcomes resulting from evidence order effects. PMID- 11477984 TI - Bispectral index monitoring in unconscious palliative care patients. AB - This pilot study looks at the clinical application of bispectral index (BIS) monitoring within the palliative care situation. Using this instrument, the level of awareness in 12 patients was tracked from the onset of unconsciousness until death, and the levels were then related to the patients' clinical state and treatment. The results give a surprising insight into the nature of unconsciousness in dying patients and the effects of treatment. The monitor proved to be a simple, effective, and acceptable method of assessing awareness in palliative care patients. A few minor problems were encountered in adapting it to the palliative care situation, but most of these were overcome as we became more familiar with the instrument. The study suggests that BIS could contribute significantly to patient care and lead to a better understanding of the dying process. PMID- 11477985 TI - A terminal care support team in a Paris university hospital: care providers' views. AB - OBJECTIVES: Following three years of a terminal care support team (TCST) activity: (a) to describe current management of terminally ill patients from the care providers' viewpoint, (b) to compare the views of head nurses and physicians about this management, and (c) to identify ways of further improving palliative care. METHODS: A questionnaire was sent to all head nurses and physicians of the hospital. RESULTS: Care-providers consider that the TCST provides therapeutic counselling, training, and psychological help to patients, staff, and families. Few care providers were reluctant to request advice in the management of their patients. Most were satisfied with the partnership with the TCST and deemed it effective. Further improvements suggested for terminally ill patient care included greater assistance from the TCST, especially for nurses, and expanding TCST activities to outpatient management and home care programs. CONCLUSION: Health care providers consider the implementation of the TCST to have been successful. PMID- 11477986 TI - Methadone: the renaissance. PMID- 11477987 TI - The development of hospice care in Arusha, Tanzania: lessons from the neighbouring states of Kenya and Uganda. PMID- 11477988 TI - Clinical outcomes in terminally ill patients admitted to hospice care: diagnostic and therapeutic interventions. AB - This study's aim was to determine the impact of medical technologies on patient comfort and survival time, through retrospective review of the clinical course, symptom profile, and illness trajectory in 102 consecutive patients (50 males and 52 females), and of diagnostic and therapeutic interventions delivered to them. The average age of males was 72.3 years and of females 73.1 years. Ninety-four patients were admitted for palliation of symptoms due to malignant disease and eight other patients for non-malignant diseases. The median survival time was 12 days. On admission, higher univariate hazard risks for survival were significantly associated with male gender, metastatic disease, and dyspnea. Higher adjusted Charlson comorbidity scores were associated with significantly decreased survival time, while de novo symptoms and diagnostic interventions were associated with lower univariate risk rates and increased survival times. Palliative therapeutic interventions were not significantly associated with increased patient survival. A multivariate analysis showed that pain, dyspnea, immobility, and adjusted Charlson comorbidity scores were independent risks for shorter patient survival times. Diagnostic interventions were significant for increased patient survival. No requests for euthanasia had been recorded, which may, in part, reflect the significant family support most of these patients had received. PMID- 11477989 TI - Keeping the hospice spirit alive in client satisfaction surveys. AB - Funding bodies and accreditation organizations now demand objective data from hospice evaluations, and quantitative measures assessing outcomes. This is despite the fact that the goals and outcomes of hospice practice are not reductionist and are not easily quantified. One alternative to this bureaucratic challenge that does not necessitate compromise to hospice ideology is to develop an assessment tool through qualitative research using phenomenological data on carers' insights with regard to hospice practice. In this article, the findings from the qualitative research that preceded the development of the assessment tool provide an insight into the key issues that carers give priority in evaluating a hospice. These findings provide valuable baseline data for understanding aspects of hospice practice that are valued by carers. The information gathered is a significant affirmation of the success of the hospice ideology and highlights the need to protect this precious philosophy. In short, it is an important statement about the need to keep the hospice spirit alive in client satisfaction. The findings from the qualitative study for the questionnaire can be interpreted as an affirmation to hospice workers that their vision and beliefs are valued and appreciated by those they serve. PMID- 11477990 TI - Multiprofessional palliative care education: past challenges, future issues. AB - Historically, health and social care professionals have described their lack of competence and confidence in many aspects of palliative care, and have recognized the need for increased educational opportunities, where new skills can be acquired and existing knowledge consolidated. Redressing these omissions has led to the development and growth of educational programs. Many of these courses have been unidisciplinary, with concomitant limitations. The adoption of a generalist approach to education has been suggested, to facilitate greater collaboration among professions and to ensure better use of resources. Course participants have greeted multiprofessional programs with enthusiasm. However, evidence of their effectiveness and impact on the delivery of palliative care to patients and their carers, although positive, remains scant. As the phenomenon of demographic aging continues, the growth of multiprofessional educational opportunities will amplify the need to scrutinize their content more closely. PMID- 11477991 TI - Constructions of dignity in end-of-life care. AB - The meaning of dignity is commonly assumed but rarely examined in palliative care. Dying with dignity often forms the basis of clinical decision making at the end of life, but is constructed differently depending upon setting and context. A discourse analysis of patient and family case studies found that relationships and embodiment were important aspects of dignity that have been neglected in the literature, although these constructions of dignity matter to dying people and their families. An understanding of these constructions can assist clinicians in providing sensitive palliative care across a range of community and medical settings. PMID- 11477992 TI - Road map for the mind. Old mathematical theorems unfold the human brain. PMID- 11477993 TI - The peak of success. The Big Bang theory clicks together better than ever. PMID- 11477994 TI - The post-genome project. PMID- 11477995 TI - Computing with light. Classical waves for pseudo quantum computing. PMID- 11477996 TI - An environmental solution. Ionic liquids may replace hazardous solvents. PMID- 11477997 TI - U.S. workers and the law. Labor rights of Americans lag behind those of other nations. PMID- 11477998 TI - The company's company. Venture capitalism becomes a new mission for the nation's spymasters. PMID- 11477999 TI - Talking gene patents. PMID- 11478000 TI - Go forth and replicate. PMID- 11478001 TI - The ice of life. PMID- 11478002 TI - Cybernetic cells. PMID- 11478003 TI - Once we were cannibals. PMID- 11478004 TI - Taming the killing fields of Laos. PMID- 11478005 TI - The do it yourself supercomputer. PMID- 11478006 TI - The trouble with turtles. PMID- 11478007 TI - Touchy-feely computing. A new mouse picks up good vibrations. PMID- 11478008 TI - Direct optometrist referral of cataract patients into a pilot 'one-stop' cataract surgery facility. AB - OBJECTIVES: To determine the feasibility of a) direct optometrist referral of patients with cataract, and b) combined assessment with same day cataract surgery ('one stop' cataract surgery). METHODS: Evaluation of 169 patients referred directly by optometrists into a pilot 'one stop' cataract surgery facility. RESULTS: Of 169 referrals, 160 patients (94.7%) were given confirmed appointments for the 'one stop' cataract service and 9 patients (5.3%) were appointed conventionally. Of 160 patients attending the 'one stop' cataract service, 154 patients (96.3%) underwent cataract surgery at the same visit, in 4 patients (2.5%) cataract surgery was indicated but deferred and in 2 patients (1.3%) cataract surgery was not indicated. The referral was supplemented with information regarding the patient's medical history forwarded by the general practitioner for 3 patients (1.8%). There were no systemic or sight-threatening complications. 151 patients (98.1%) achieved a visual acuity of 6/12 or better at a mean of 31 days post-operatively. CONCLUSION: Optometrists can accurately predict the need for cataract surgery and refer directly into a pilot 'one stop' cataract surgery facility, without the need for general practitioner involvement. 'One stop' cataract surgery is feasible; benefits to the patient include the abolition of the need to visit the general practitioner for consultation and referral, and the hospital for pre-assessment. PMID- 11478009 TI - Post-operative pain in needlescopic versus conventional laparoscopic cholecystectomy: a prospective randomised trial. AB - BACKGROUND: Needlescopic cholecystectomy (NC) utilises instruments and ports smaller than 3 mm in diameter compared with the 5 mm ones used in conventional laparoscopic cholecystectomy (LC). Post-operative pain control and recovery has been thought to be superior in NC, when compared with historical controls with LC, but has not been proven in a prospective fashion. PATIENTS AND METHODS: A prospective randomised trial of NC versus LC for patients with symptomatic gallstone disease, with standardisation of post-operative analgesia and daily assessment of post-operative pain, using a 5-point visual analogue scale. RESULTS: There were 64 eligible patients randomised into NC (28) and LC (36). Four patients who had NC were converted to LC due to technical problems. Another three and six patients from the NC and LC groups, respectively, had conversion to open surgery. Post-operative pain scores were low in both groups. Mean pain scores for those with successful NC and LC were: 1.24 versus 1.43 for the day of operation (P = 0.49), 0.86 versus 0.83 for the first day post-operatively (P = 0.92) and 0.75 versus 0.81 for the second post-operative day (P = 0.87). The mean number of intra-muscular analgesic injections required were 0.76 versus 0.83 after NC and LC, respectively (P = 0.93). There were no significant differences between the two groups in the time taken to return to feeding, eating a normal diet and discharge from hospital. CONCLUSION: There is no advantage of NC over LC in terms of post-operative pain or recovery. Nevertheless, NC can be performed safely and expediently and has an excellent cosmetic outcome and high patient acceptability. PMID- 11478010 TI - The delivery of surgical cleft care in the United Kingdom. AB - BACKGROUND AND PURPOSE: A national survey of cleft teams was undertaken as part of the Clinical Standards Advisory Group investigation of the current status of cleft care in the United Kingdom (UK). METHODS: Fifty-seven cleft teams were identified, of which 90% responded to the survey. MAIN FINDINGS: Nine cleft teams had been established since 1992. Only one region, Northern Ireland, had a centralised cleft service and, despite 82% of teams having databases, only four were able to produce corroborated evidence of receiving at least 30 annual new referrals during 1995. There was a wide variation in the facilities provided by individual cleft teams--only six teams were able to provide all of the key facilities recommended by the Royal College of Surgeons Steering Group on cleft lip and palate. Facilities such as antenatal and neonatal counselling, protocols for record keeping and long-term treatment were similar for high and low volume teams. High volume teams were more likely to have established links with a full range of specialities including psychology, clinical genetics and paediatrics than low volume teams. CONCLUSION: A national survey of cleft services has demonstrated a need for reorganisation. This is now in process and once established will require continual monitoring and assessment. PMID- 11478011 TI - Modern management of head injuries. AB - Management of the head-injured patient is designed to prevent secondary injury and to provide the neurosurgeon with a live patient who has some hope of recovery. This review sets out the background essentials for the non-neurosurgeon dealing with the initial care of a head-injured patient. PMID- 11478012 TI - Surgical treatment of varicose veins. PMID- 11478013 TI - Dendritic cells (II): Role and therapeutic implications in cancer. AB - The potential to harness the effectiveness and specificity of the immune system underlies the growing interest in cancer immunotherapy. One such approach uses bone marrow-derived dendritic cells (DCs), phenotypically distinct and very potent antigen-presenting cells, to present tumour-associated antigens (TAAgs) and, thereby, generate tumour-specific immunity. Support for this strategy comes from animal studies that have demonstrated that DCs, when loaded ex vivo with tumour Ags or pulsed with peptides and administered to cancer-bearing hosts, can elicit T cell-mediated cancer destruction. These observations have led to clinical trials designed to investigate the immunological and clinical effects of Ag-pulsed DCs administered as a therapeutic vaccine to patients with cancer. In the design and conduct of such trials, important considerations include Ag selection, methods for introducing TAAgs into MHC class I and II processing pathways, methods for isolating and activating DCs, and route of administration. Although current DC-based vaccination methods are cumbersome and complex, promising preliminary results from clinical trials in patients with malignant lymphoma, melanoma, and prostate cancer suggest that immuno-therapeutic strategies, that take advantage of the unique properties of DCs, may ultimately prove both efficacious and widely applicable treatment in patients with cancer. PMID- 11478014 TI - The hepatic artery: a reminder of surgical anatomy. AB - This study was carried out to document the anatomy of the hepatic artery with the purpose of reminding surgeons of the need for this essential knowledge in order to practice safe hepatobiliary surgery. Repeated surgical mistakes on patients referred to our unit prompted the study. One hundred and eighty consecutive livers procured for transplantation was studied, and the anatomy drawn immediately after dissection. The left hepatic artery arose from the left gastric artery in 15%, and either the splenic, gastroduodenal artery or the aorta in 4% of cases. The right hepatic artery arose from the superior mesenteric artery in 15%, the gastroduodenal, right gastric artery or aorta in 10% of cases. There was a major variation of the coeliac axis in 9% of cases studied. Overall, there was an abnormality in 43% of dissections: 48% were multiple and 27% had more than two vascular variations. A constant pattern of abnormalities occurred in the anatomy of the hepatic artery. Realisation of this vascular pattern should make identification of the anatomy easier. When there is one vascular variation, there is a high chance of there being multiple variations. PMID- 11478015 TI - Clinical audit: can we improve further? AB - Clinical audit is an important tool for comparing one's practice against existing standards. The authors have analysed the Junior Doctor's understanding of audit by performing a survey and questionnaire from 146 trainees, SpRs and SHOs from nine hospitals. The study showed that 107 (72.8%) performed audit and among those who performed audit, 52 (48.6%) experienced difficulty in obtaining data. This study highlights the importance of support needed to encourage the junior doctors to participate in audit programmes. PMID- 11478016 TI - Carotid artery aneurysm secondary to cystic medial necrosis. AB - Carotid artery aneurysm secondary to cystic medial necrosis is a rare clinical entity. We report a 59-year-old Chinese male patient who presented with a pulsatile right neck swelling for 2 months. Partial resection of the aneurysm with primary anastomosis of the internal carotid artery was performed. Histopathological examination of the aneurysmal wall demonstrated cystic degeneration of the media with accumulation of glycosaminoglycan material, consistent with the features of cystic medial necrosis. The pathogenesis of carotid artery aneurysm secondary to cystic medial necrosis is discussed. PMID- 11478017 TI - Bilateral gynaecomastia as the primary complaint in hyperthyroidism. AB - Association of gynaecomastia with hyperthyroidism is uncommon but has been well documented in the past. Gynaecomastia in a patient with hyperthyroidism rarely presents as a primary complaint. When this occurs, it may present a diagnostic challenge to the clinician. We present the case of a patient who was referred initially to the breast clinic with bilateral gynaecomastia. Hyperthyroidism was subsequently confirmed and treated; gynaecomastia regressed with return to the euthyroid state. PMID- 11478018 TI - Muir-Torre syndrome. AB - Muir-Torre syndrome is a rare autosomal dominant condition in which multiple primary malignancies occur together with a sebaceous gland tumour. Early recognition of the syndrome in patients with sebaceous gland tumours should facilitate early detection of subsequent malignancies if the patient with entered into appropriate screening programmes. A case occurring in Scotland is described and implications for management, screening for members of the family are discussed. PMID- 11478019 TI - Obscure gastrointestinal bleeding due to carcinoid tumours of the small bowel: problems in diagnosis and localisation. AB - Patients with obscure gastrointestinal bleeding are often difficult to manage and frequently undergo extensive investigations. The diagnosis is particularly difficult when the bleeding arises from small lesions in the small bowel that is not easily accessible for direct visualisation. We report two patients with obscure gastrointestinal bleeding from carcinoid tumours of the small bowel, highlighting the problems in diagnosis and localisation. PMID- 11478020 TI - Laparoscopic port site recurrence in the absence of intra-abdominal disease. AB - A case is reported of a patient who presented with recurrent tumour at the site of a laparoscopy port where there had been no evidence of intra-abdominal tumour. Possible mechanisms of recurrence are postulated including a haematogenous spread of the primary tumour and the implantation at the port site. PMID- 11478021 TI - Transient hemi-diaphragmatic paralysis following neck surgery: report of a case and review of the literature. AB - Diaphragmatic paresis following trauma to the phrenic nerves is a rare complication after neck surgery. The resulting elevation of the ipsilateral hemi diaphragm is diagnosed on post-operative chest radiography and may be confirmed by ultrasound or fluoroscopy. When unilateral, this may lead to respiratory, cardiac or gastrointestinal symptoms and atelectasis and pulmonary infiltrates on radiography. If nerve damage is bilateral a period of ventilation may be required. PMID- 11478022 TI - A modification to aid open tracheostomy. PMID- 11478023 TI - A modification to aid open tracheostomy. PMID- 11478024 TI - A modification to aid open tracheostomy. PMID- 11478025 TI - A modification to aid open tracheostomy. PMID- 11478026 TI - A modification to aid open tracheostomy. PMID- 11478027 TI - A modification to aid open tracheostomy. PMID- 11478028 TI - Altered cardiac function. PMID- 11478029 TI - Repair of incisional hernias. PMID- 11478030 TI - Facial nerve palsy following intra-oral surgery performed with local anaesthesia. PMID- 11478031 TI - Facial nerve palsy following intra-oral surgery performed with local anaesthesia. PMID- 11478032 TI - [Genetics of renal tumors]. AB - The quintessence of malignant transformation is the genetic alteration of the tumor progenitor cell, i.e. somatic mutation. The genetic change appearing at chromosome and/or gene level results in the disturbance of the balance of cell proliferation and differentiation. In solid tumors, including renal tumors, the basic genetic mechanism proved to be the loss of function of a specific gene pair caused by loss of the particular chromosome or chromosomal region (monosomy, deletion) or by mutation of the gene. In the well studied Wilms' tumor-aniridia syndrome the distal part of 11p13 region is deleted. The responsible gene is the WT-1 tumor suppressor gene, a Zn finger type transcription factor. In the majority of cases the mutation of this gene leads to the tumor formation without cytogenetically detectable deletion. For manifestation of the tumor the functional damage of both alleles is needed. In other histological types of renal tumors a great variation of chromosome losses and gains, as well as translocations can be identified. In Wilms tumor of embryonic origin, tumor suppressor genes located on the short arms of chromosomes 16 and 17 play a role in the pathogenesis. Besides, the significance of abnormal genomic imprinting of IGF2 and H19 genes located on 11p15 has also been confirmed. If a part of the embryonic cells do not regress, they may develop to papillary carcinoma together with the appearance of trisomies of chromosomes 7 and 17 and loss of Y. In the transformation process from papillary adenoma to carcinoma, duplication of several chromosomal regions is involved (3q+, +8, +12, +16, +20). The origin of renal carcinoma developing from normal nephron cells is associated with a deletion of 3p and 5q+, while during the progression of the disease further variable chromosome losses appear (6q-, 8p-, 14q-, -9). Tumor-specific cytogenetic and molecular genetic changes confirm the morphological classification of epithelial renal tumors pointing at the relation of the various entities or their independence. Based on cytogenetic alterations, a sequential predictive model of renal tumors can be developed. Individual types, together with joining and sequential appearance of aberrations are in line with the multistep mechanism of carcinogenesis. At the same time, the specific cytogenetic and molecular genetic changes confirm the diagnosis, provide further information about the histological type and progression of the disease. In hereditary cases, the members of the family at risk can be identified by recognizing the possibly associating clinical symptoms and/or by detecting the constitutional mutation of the gene using molecular genetic methods. Consequently, the genetic study of renal tumors plays important role not only in diagnosis and choosing adequate therapy but also in prevention of the disease. PMID- 11478033 TI - [Left ventricular hypertrophy: pathogenesis, diagnosis and therapy]. AB - The life expectancy of hypertonic patients is basically determined by the complications of their disease. The increasing hypertension induces compensated processes in cardiovascular system, however, these alterations further worsen the mortality rates. It is evident that left ventricular hypertrophy (LVH) should be prevented. If it is the case, it has to be moderated among the possible circumstances. To regression of LVH produced by hypertony the decrease of hypertension is not enough, while the process is rather complex. Because the structural changes in the interstitial tissue followed by the alterations of heart myocytes hypertrophy, hyperplasy and apoptosis of myocytes, as well as the interstitial and perivascular fibrosis, coronary remodelling and necrosis in the consequence of insufficient blood circulation can be widely observed. The individual prognosis of LVH can be determined by echocardiography, while the concentric remodelling the development of excentric and concentric hypertrophy is strictly parallel with the increase in the mortality and morbidity rates. According to the evidences the treatment consists of life mode controlling and suitable drug therapy. First line medicines are the ACE-inhibitors and indapamides, later on these drugs can be further combined with calcium antagonists and beta-blockers, if necessary. PMID- 11478034 TI - [Review of the various methods investigating the CNS-complications of hypertension]. AB - To summarise the structural and functional changes of the brain, caused by chronic hypertension and overview the studies performed on hypertensive patients by computer tomography, magnetic resonance imaging, transcranial Doppler ultrasound, positron emission tomography and single photon emission tomography. The definitive lesions--global or local atrophy, lacunar infarcts--can be sensitively detected by different morphological methods, whereas the hemodynamical alterations can be observed by the functional techniques. Mild cognitive deficits could be diagnosed in the early stage of the disease with different neuropsychological testing. Positron emission tomographic and single photon emission tomographic investigations could not be used routinously on hypertensive patients. The authors--based on own experiences--suggest the combined use of transcranial Doppler and neuropsychological testing, because these techniques are sensitive enough to detect early, subclinical abnormalities. PMID- 11478035 TI - [Open heart surgery with extracorporeal circulation during pregnancy]. AB - The incidence of heart disease in pregnancy has been gradually falling during the last three decades. Cardiopathy still remains a prominent cause of maternal and fetal morbidity and mortality. Most patients know about their heart disease long before conception, even though the potential risk factors of deteriorating cardiac function during pregnancy are generally not emphasized. These women when pregnant may develop heart failure due to the increased cardiorespiratory requirements. When medical therapy proves insufficient heart surgery becomes mandatory to save the patient's life. The pregnant state is not optimal for cardiac surgery as the principle interest of the mother and the fetus is different. We report on two pregnant patients who underwent unavoidable heart surgery with cardiopulmonary bypass and review the literature regarding the optimal management of open-heart operation in pregnancy aiming to decrease the feto-maternal mortality. The successful outcome of the cardiac surgery on pregnant women is determined by the severity of the preexisting disease, the surgical techniques, and the circumstances of the cardiopulmonary bypass. The best possible results can be achieved by providing preconceptional counseling for the cardiopathic patients regarding the relation between the preexisting risk factors and the adverse maternal and neonatal outcome. When heart surgery is mandatory in pregnancy the careful technical precautions and the continuous cardiotocography help to minimize fetal complications during the cardiopulmonary bypass (CPB). PMID- 11478036 TI - [Acute pancreatitis with few symptoms and extensive necrosis]. AB - The authors present a particular case of an acute pancreatitis. The disease developed in a young male patient following cholelithiasis and cholecystectomy. The inflammation affected the outer layers of the pancreas as a mantle and it caused widespread fat necrosis. Necrectomy for septic state was conducted to improve the condition, but the patient died of pulmonary embolism. Unexpectedly big necrotic areas of fat necrosis and abscess were found at autopsy. PMID- 11478037 TI - [Reflections about the relationship of science and art]. AB - Science is dealing with the nature, the human beings and the society, and aims to explore the laws of their existence. It uses universal scientific methods, by taking all known laws of nature into consideration. It is free of subjectivity and is guided by a high degree of consciousness. In the author's view, the goal of science is to create a balance between man and nature by exploring the rules of the universe. Art, on the other hand, carries a message about the man and the world, which originates in both emotion and intuition. It intends to impress our feelings and wishes to entertain, create pleasure or make us accept its message. One might wonder about the nature of relationship between art and science. Do we scientists waste our valuable time when dealing with art? Furthermore, is it of any use for artists to deal with science? Ever since the ancient times, scientists have been highly appreciated. Artists, however, used to belong to lower social classes up until the 18th century. Still, the commercial and political life of the Middle Ages were greatly influenced by the guildes, where artists as craftsmen belonged to. Art and science have always been interconnected, although their contents and messages kept changing during the centuries. In the 5th century, sciences were listed among the "septem artes liberales", the "seven free arts". When comparing the creative process of art and science, we might find similarities and differences, some of which will be discussed in the paper. Both research and clinical profession demands devoted work. One of the most valuable form of stress reduction and relaxation is the enjoyment or practice of art. Engagement in art as a form of hobby widens our horizon which in turn stimulates professional work. We might as well agree with the wise saying: Without hobby, one can neither relax nor concentrate. PMID- 11478038 TI - Functional ligamentous anatomy of the trapezium and trapeziometacarpal joint (gross and arthroscopic). AB - A detailed description of the ligamentous anatomy of the thumb basal joint is presented from a gross, arthroscopic, and functional point of view. All 16 ligaments that stabilize the trapezium and trapeziometacarpal (TM) joint are discussed in association with their presumed functional relevance regarding trapezial and trapeziometacarpal joint stability. Geometric and mathematical models are introduced to help describe and support anatomic observations with regard to ligament function. The dorsoradial ligament appears to be an important stabilizer of the TM joint against dorsally directed forces. The trapeziotrapezoid, trapezio-II metacarpal, and trapezio-III metacarpal ligaments appear to function as tension bands to help support the trapezium against cantilever bending forces imparted onto the trapezium by the thumb metacarpal. PMID- 11478039 TI - Thumb kinematics and their relevance to function. AB - With its sophisticated motion and prehensile ability, the TM joint is essential to hand function. The bony anatomy and the musculotendinous forces across the joint allow this advanced function. The elaborate ligamentous system provides stability. The high forces transmitted during routine pinch and grasp would not be possible without this complex arrangement. PMID- 11478040 TI - Pathomechanics of the thumb trapeziometacarpal joint. AB - As the human thumb climbed the evolutionary ladder, the trapeziometacarpal joint was provided greater mobility at the expense of decreased bony stability. Soft tissue restraints were left to provide the primary stabilization of the thumb basal joint complex. Both degenerative and traumatic conditions compromise the function of these ligamentous structures and result in translational instability of the joint surfaces and secondary arthritic disease. Primary idiopathic osteoarthritis, Bennett's fracture-dislocation, and metacarpophalangeal hypermobility all impart a disturbance in biomechanics while extension metacarpal osteotomy, ligament reconstruction, and imposed metacarpophalangeal flexion mitigate these effects and improve trapeziometacarpal kinematics. As a compact joint amenable to en bloc resection, the trapeziometacarpal complex affords a unique opportunity to study the inter-relationship between biology and biomechanics in the pathogenesis of degenerative joint disease. PMID- 11478041 TI - Clinical assessment of the thumb trapeziometacarpal joint. AB - Osteoarthritis of the basal joint of the thumb causes pain and disability for a huge segment of the adult population, particularly women. Radiographically, there is a spectrum of disease that has been staged according to severity of involvement of the TM and scaphotrapezial joints. The staging system proposed by Littler and Eaton is used most widely. The severity of clinical symptoms does not necessarily correspond with the radiographic stage of disease, however, so decisions about treatment are predicated upon the notion that we "treat patients, not x-rays." Treatment is based upon the extent to which the pain and functional limitations caused by the disease impact upon the patient's activities of daily living. The evaluation of basal joint disease has been reviewed and modalities of treatment outlined. Conservative treatment includes splinting, nonsteroidal anti inflammatory drugs, thenar intrinsic strengthening exercise, and corticosteroid injection. Failure of conservative treatment and unremitting pain are indications for basal joint reconstruction by arthroplasty, osteotomy, or arthrodesis. Staging of basal joint disease provides a rationale for selecting the appropriate surgical procedure for a particular patient. PMID- 11478042 TI - Treatment of Eaton stage I trapeziometacarpal disease. Ligament reconstruction or thumb metacarpal extension osteotomy? AB - Although the cause of TM joint hypermobility may be multifactorial, translational laxity and instability appear to be related to degeneration of the beak ligament. Indeed, the forces experienced at the normal TM joint with grip and pinch are magnified several fold and are concentrated in the palmar aspect of the joint. The TM stress test used in preoperative assessment of thumbs with hypermobile TM joints reflects nonphysiologic dorsal translation and correlates with histologic evidence of detachment of the metacarpal origin of the beak ligament. Ligament reconstruction of the painful TM joint reflects the importance of the beak ligament in providing stability and in limiting dorsal translation of the metacarpal that normally occurs during dynamic pinch activity. Metacarpal extension osteotomy, by contrast, shifts mechanical loading at the TM joint more dorsally and redirects force vectors. Its precise role in the treatment of TM disease has long remained ill-defined because of a lack of correlation between preoperative disease status and postoperative functional result. It appears now, however, that for stage I disease, metacarpal extension osteotomy may result in satisfactory pain relief and improved grip and pinch strength--at least in the short term. Indeed, extension osteotomy may be an efficacious alternative to ligament reconstruction, but long-term follow-up and additional clinical studies will be helpful in shedding additional light on this topic. PMID- 11478043 TI - Ligament reconstruction tendon interposition arthroplasty for basal joint arthritis. Rationale, current technique, and clinical outcome. AB - The literature unequivocally supports the LRTI arthroplasty. Indeed, thumb stability, pain relief, and improvements in strength are the expected outcomes. Although some investigators believe that ligament reconstruction is not necessary, trapezium excision alone, or in combination with facial or tendon interposition, is less likely to provide long-term stability or restore satisfactory pinch and grip strength. Some hand surgeons may be apprehensive about harvesting the entire width of the FCR tendon because of fear that wrist function may be impaired or that a larger bony channel in the metacarpal might result in fracture. In that light, Coleman and the author recently reported the results of a prospective evaluation that rather convincingly showed there was no morbidity accompanying harvest of the entire FCR tendon, from the standpoint of wrist strength or endurance. Furthermore, technical modification by which the end of the FCR is tapered or trimmed obviates the need for an excessively large bony channel through the metacarpal. Preliminary pin fixation of the metacarpal, with its base suspended at the level of the index CMC joint, in the fisted position, is still recommended to allow accurate tensioning of the new ligament, and protection in the early postoperative period. Proximal migration of only 13% at an average of 9 years after surgery may very well reflect the value of this particular practice. In conclusion, attention to technical detail and compliance with the postoperative therapy program, in the author's opinion, are intricately related to the favorable outcomes he has observed during 5 years of seeing Burton perform the LRTI arthroplasty and more than 6 years of performing the procedure in his own practice. PMID- 11478044 TI - Trapezio-metacarpal arthritis. Trapezium excision and ligament reconstruction not including the LRTI arthroplasty. AB - For trapeziometacarpal arthritis, trapezium excision and ligament reconstruction that is distinct from the ligament reconstruction tendon interposition (LRTI) arthroplasty has a distinct role. Emphasis is placed on the use of the abductor pollicis longus for tendon interposition and "suspensionplasty" for the arthritic thumb carpal metacarpal joint. The advantages of the abductor pollicis longus techniques as originally advocated by Thompson, and modified by Diao, are reviewed. The surgical technique for this procedure, clinical results and biomechanical cadaver analysis comparing abductor pollicis longus suspensionplasty done with two techniques, as they compare to the LRTI procedure, are included. The abductor pollicis longus suspensionplasty is an excellent treatment choice both for index procedures for carpometacarpal (CMC) thumb arthritis, and for salvage of the failed thumb CMC arthroplasty. PMID- 11478045 TI - Treatment of arthritis of the trapeziometacarpal joint with trapeziectomy and hematoma arthroplasty. AB - Trapezial excision is a simple and effective method of treating painful TM arthritis. Recent studies, as well as the authors' experience, suggest that the results of this procedure, in the short-term, provide equivalent pain relief compared with other, more complex techniques requiring tendon interposition and ligament reconstruction. Trapezial excision and hematoma arthroplasty is now the authors' procedure of choice for treating painful arthritis at the base of the thumb, regardless of Eaton stage. Long-term prospective comparison of trapezium excision alone and LRTI arthroplasty may help determine whether the additional strength gains accompanying the latter are associated with increased patient satisfaction, in high-demand patients. PMID- 11478046 TI - Treatment of trapeziometacarpal arthritis with silastic and metallic implant arthroplasty. AB - Implant arthroplasty was once viewed as a feasible and effective trapeziometacarpal joint disease. Silastic implants are now used rarely because of concerns of implant failure and particulate synovitis. Metallic implants, including total joint prostheses and those utilizing a spacer concept, have been used with some success. Indications, technical consideration, and potential complications of these implants are discussed. PMID- 11478047 TI - The role and implementation of metacarpophalangeal joint fusion and capsulodesis: indications and treatment alternatives. AB - Successful surgical reconstruction of the basilar thumb joint requires a thorough knowledge of the longitudinal intercalated collapse that may occur with this condition. Evaluation of the hyperextended thumb MCP joint and appropriate treatment are instrumental to successful basal joint reconstruction. Stabilization of the MCP joint helps maximize return of a pain-free, efficient pinch mechanism. A useful guide to specific reconstructive methods for the MCP joint is presented in Table 1. PMID- 11478048 TI - The treatment of trapeziometacarpal arthritis with arthrodesis. AB - In this article, the authors discussed the indications for TM arthrodesis, the surgical approach, the types of fixation, expected outcome, and complications. Fusion of the thumb TM joint allows maintenance of pinch and grip strength and provides relief of pain, but limits thumb mobility. An important factor in the success of the arthrodesis is correct thumb position. Trapeziometacarpal joint arthrodesis is advantageous over soft tissue interposition when grip and pinch strength are to be maintained. After TM fusion, however, there are increased stresses across the peritrapezial joints that can cause laxity, pain, and arthritis. Treatment of this may require additional surgical intervention. Treatment of TM arthritis with soft tissue interposition has the advantages of pain relief and increased mobility, but pinch and grip strength are reduced to approximately 75% of normal and rate of reoperation is minimal. The authors recommend TM arthrodesis in the young active person with arthritis limited to the TM joint in whom strong pinch and grip are required. Arthrodesis of the TM joint is safe and predictable and has good subjective and objective results. PMID- 11478049 TI - Implant arthroplasty of the metacarpophalangeal joint of the thumb. AB - Thumb metacarpophalangeal arthritis may affect overall hand function more than expected because pain and instability compromise pinch and grip strength. Implant arthroplasty represents one option in the treatment of deformity, destruction, and instability at the metacarpophalangeal joint level. Appropriate use of implant arthroplasty requires careful evaluation of the overall deformity and the status of the interphalangeal joint. If the interphalangeal joint is likely to require fusion--either simultaneous with metacarpophalangeal surgery or subsequently--then implant arthroplasty may be the best treatment option for the metacarpophalangeal joint. PMID- 11478050 TI - Surgical treatment of the rheumatoid thumb. AB - Rheumatoid arthritis commonly affects the thumb. Deformity does not require surgical intervention unless pain is present or a functional deficit exists, but if bony erosion develops and surgical treatment may be compromised in the future, earlier intervention may be indicated. When the pathogenesis and pathoanatomy of the impaired rheumatoid thumb are appreciated, and appropriate treatment is selected, surgical intervention is likely to provide a favorable outcome for the patient. PMID- 11478051 TI - Disorders of the thumb sesamoids. AB - The subsesamoid joints of the thumb are a common site of arthritis, but their small size makes diagnosis of disorders challenging. The sesamoid and subsesamoid joints may also be injured acutely with the volar plate complex during hyperextension injuries, and may sometimes produce mechanical dysfunction. Simple excision of chronically painful sesamoids provides excellent relief. PMID- 11478052 TI - The surgical treatment of scaphotrapeziotrapezoid osteoarthritis. AB - The incidence of scaphotrapeziotrapezoid osteoarthritis increases during the aging process. This article discusses the surgical options available to treat this condition. PMID- 11478053 TI - The role of arthroscopy in evaluating and treating trapeziometacarpal disease. AB - Thumb trapeziometacarpal arthroscopy offers unique opportunities for improving evaluation and treatment in patients with basal joint disease. For patients with early disease, arthroscopic synovectomy and electrothermal shrinkage of the capsule can provide symptomatic improvement. For patients with more advanced disease, arthroscopic hemitrapeziectomy and complete trapeziectomy can be performed, combined with electrothermal shrinkage of the anterior oblique ligament. This article reviews the surgical technique and the early follow up of this rapidly advancing, minimally invasive technique for treatment of trapeziometacarpal disease. PMID- 11478054 TI - Current concepts regarding pharmacologic treatment of rheumatoid and osteoarthritis. AB - Treating patients with osteoarthritis (OA) and rheumatoid arthritis (RA) remains challenging; however, new agents offer the chance for an improved quality of life. As an alternative to traditional nonsteroidal anti-inflammatories, cyclooxygenase-2 inhibitors provide pain relief for OA and RA patients with possible fewer side effects. Otherwise, OA patients may opt for topical agents, injections, or supplements. Rheumatoid arthritis research has led to an improved understanding of the inflammatory cascade and an appreciation of the early tissue destruction. A new treatment philosophy has thus emerged along with the development of new biologic agents; the latter, along with combination therapy and a new disease modifying antirheumatic drug, leflunomide, have greatly expanded the chances for disease control in RA patients. PMID- 11478055 TI - Nitric oxide and glioma: a target for novel therapy? AB - The biological activities of nitric oxide (NO) include vasodilatation, inhibition of platelet aggregation, neurotransmission, neural plasticity, and modulation of inflammatory and immunological functions. NO synthase (NOS), which is the enzyme that produces NO, has been detected in resected human glioma specimens, and both human and rodent glioma cell lines. NO production in gliomas can alter several important pathophysiological processes, such as local host immune response, tumour cell apoptosis, tumour invasion/metastasis, free radical injury to tumour cells and adjacent normal brain tissues, tonic vasodilatation of tumour vessels, vascular permeability and neovascularization. Recently, some therapeutic strategies for gliomas using NO manipulation have been proposed, and evaluated both experimentally and indirectly in preliminary clinical trials. These include NO manipulation designed to modify tumour cell oncogenesis, tumour blood flow and disposition of anti-cancer drugs in tumour tissue. This review will discuss the biological role of NO in the central nervous system and gliomas and its current and future possibilities in neuro-oncology. PMID- 11478056 TI - Do adjustable shunt valves pressure our budget? A retrospective analysis of 541 implanted Codman Hakim programmable valves. AB - Based on a retrospective study in which a total of 541 Codman Hakim Programmable Valves (CHPV) were implanted in 477 patients over a 6-year period, this cost analysis was performed. By using a valve with an adjustable opening pressure, valve exchange to alter the opening pressure and surgical evacuation of subdural haematomas and hygromas can be avoided. Dividing the added cost for using the CHPV by the cost of implanting a non-programmable Hakim valve results in 105.8 valve exchanges, which would have had to be avoided to break even financially. On 107 occasions a valve was adjusted by a magnitude of 50 mmH2O or more. This, if an adjustment of that magnitude is said to correspond to a valve exchange, is sufficient to break even. The analysis suggests that the extra cost of the valve is outweighed by the ability to adjust the opening pressure setting non invasively. PMID- 11478057 TI - Endovascular approach to treatment of indirect carotico-cavernous fistulae. AB - The purpose of the study was to assess the cure rate in patients with indirect carotico-cavernous fistula (CCF) treated by transvenous embolisation via the inferior petrosal sinus (IPS) pathway or the superior ophthalmic vein (SOV). Twelve fistulae in 11 patients were treated by transvenous embolisation, eight patients via the IPS and four fistulae in three patients via the SOV. Angiographic cure was defined as complete obliteration of the fistula and clinical cure as total resolution of signs and symptoms. Complete cure was achieved in eight patients with nine fistulae embolised transvenously. Five fistulae were approached via the IPS and four via the SOV. Our preferred method for treatment of indirect CCF is the transvenous route. The IPS approach is technically easier and has fewer potential risks than the SOV approach. However, if the IPS is not patent the SOV can provide good alternative access to the cavernous sinus. PMID- 11478058 TI - Endoscopic treatment of prepontine arachnoid cysts. AB - Prepontine (or suprasellar) arachnoid cysts are uncommon in clinical practice and experience in their management may therefore be limited. Symptomatic cysts usually present with features of hydrocephalus due to obstruction of the third ventricle and aqueduct, and occlusion or partial obstruction of both foramina of Monro. Several treatment techniques have been used including stereotactic aspiration, microsurgical excision and shunting, but the best method of treatment remains unclear and the role of endoscopy is not yet established. We report our experience in a series of seven patients who had endoscopic treatment for prepontine arachnoid cysts; five were children under 15 years old who presented with delayed development and/or enlarged heads. The two adult patients, both of whom had insertion of shunts as children, presented with headache and vomiting due to shunt blockage. All patients improved following endoscopic cyst fenestration. There was no operative morbidity and there have been no relapses to date. Endoscopic fenestration of prepontine arachnoid cysts appears to be an effective method of treatment. PMID- 11478060 TI - Lymphocytic and granulocytic hypophysitis: a single centre experience. AB - Lymphocytic and granulocytic hypophysitis are rare pituitary masses. A series of five dealt with at a single centre is presented and discussed. Retrospective analysis of pathology records revealed the cases. Of pituitary mass lesions dealt with in this centre, 0.8% have the diagnosis. Five females presented, one pregnant at the time of presentation, three presenting with signs and symptoms of panhypopituitarism, and two with visual problems. All were treated by surgical means. All had good postoperative visual function, but all were panhypopituitary. Follow-up was for a mean of 4 years and 1 month. This rare condition should be treated by surgical means to obtain a diagnosis. Further treatment of residual tumour can be with corticosteroids. Good visual function should be expected, but so should permanent loss of pituitary function. PMID- 11478059 TI - Evaluation of spontaneous intracranial hypotension: assessment on ICP monitoring and radiological imaging. AB - We describe two recent cases of spontaneous intracranial hypotension. A 38-year old woman developed a severe postural headache. Magnetic resonance imaging (MRI) showed marked dural enhancement. Histopathological findings of dural biopsy showed numerous dilated vessels in the dura, rather than hypertrophic change. Lumber CSF pressure was 5 cmH2O and RI cisternography suggested CSF leakage. A 58 year-old woman with postural headache and vertigo had bilateral subdural haematoma associated with diffuse dural enhancement on MRI. Lumber CSF monitoring confirmed persistent low pressure ranging from 0-5 cm H2O. MRI myelography revealed multiple CSF pouches along the whole spinal axis. CSF leakage was demonstrated on Radioisotope (RI) cisternography. Both cases described in this report were diagnosed as spontaneous intracranial hypotension caused by CSF leakage from spinal meningeal diverticula and were successfully treated by intravenous Factor XIII administration. PMID- 11478061 TI - Prognostic significance of the endothelial surface in low-grade resected oligodendrogliomas. AB - The importance of angiogenesis as a prognostic factor in brain tumours has recently been reported. In this study, we analysed the long-term prognostic significance of a morphometric score expressing the endothelial area for every 1000 tumour cells, in tumour tissue from 26 patients with a low-grade oligodendroglioma that has been treated surgically and irradiated, and has a MIB 1 labelling index (MIB-1 LI) of less than 1%. In each tumour, a vascular endothelial surface index (VESI) was determined as the CD-34 immunostained endothelial area in micron 2 per 1000 tumour cells. Patients with a VESI of less than 15 (n = 12) showed a survival at 5 and 10 years of 100 and 71%, respectively, versus a survival of 50 and 0% for patients presenting a VESI greater than 15 (n = 14); p < 0.05). Our present findings suggest the usefulness of VESI as a long-term prognostic pathological factor in low-grade oligodendroglioma. PMID- 11478062 TI - Plate and screw technique for advancement of the supraorbital bar in surgery for craniosynostosis: a preliminary technical report. AB - An alternative plate and screw method of advancement of the supraorbital osteotomy bar in cranial remodelling surgery for craniosynostosis is described. The lateral orbital rim is transversely cut and using miniplates, screws are implanted into its advanced cut ends. Appropriate screws and miniplates are used to suit the size of the orbital rim in patients of different age groups. The technique was used in 12 cases and the authors have found the results extremely gratifying. It offered the advantage of providing a firm, secure and desired advancement, and height of the supraorbital bar maintaining a smooth lateral rim contour. The procedure was technically relatively straightforward. Further observation is warranted to determine whether the stabilization will last into adulthood or if there will be any problem related to the metal screws and plates. PMID- 11478063 TI - Endoscopically assisted closure of an anteriorly based spinal pial arteriovenous fistula. AB - The intraoperative use of an endoscope to localize an anteriorly based spinal pial arteriovenous fistula with minimal cord retraction and vessel manipulation is described. PMID- 11478064 TI - Transnasal intracranial entry of a flying wire fragment. AB - A 7-year-old boy was playfully revolving a partly insulated electric wire which accidentally struck a bamboo pole in its path. He soon found that his left nostril was hit and blood trickled from the site. Unsuspected on investigation, a small fragment of the wire was found within the brain. The sequence of events is reported. PMID- 11478065 TI - Hearing loss after surgery of ruptured cerebral aneurysm. AB - Only a few cases have been reported of hearing impairment after various procedures that result in loss of cerebrospinal fluid (CSF). We experienced eight cases of hearing loss following surgery for ruptured cerebral aneurysm. Angiography revealed that the aneurysm was located at the middle cerebral artery bifurcation in two cases, anterior communicating artery in four cases, posterior communicating artery in one case, and superior hypophyseal artery in one case. All of them were relatively far from the hearing structures. The aneurysm was clipped easily after drainage of CSF for slack brain. Pure-tone audiometry revealed unilateral or bilateral hearing loss that not only affected the low, but all frequencies. Completely normalized hearing was found in five patients at 3-6 months follow-up, whereas two cases noticed a partial loss and one made no recovery. We speculate that the explanation of hearing loss following aneurysm surgery results directly from diminished CSF pressure by the loss of CSF during operation. This decrease is transmitted to the perilymph via a patent cochlear aqueduct (CA) producing the release of perilymphatic fluid in the subarachnoid space. It is possible that a haemolabyrinth caused by an inflow of subarachnoid haemorrhage through CA could be an additional factor in the induced impaired hearing. PMID- 11478066 TI - Progressive ulnar neuropathy caused by delayed migration of a foreign body. AB - Foreign bodies in the hand can cause a variable degree of acute or delayed injuries to the important structures. We report a rare case of a progressive ulnar neuropathy caused by delayed migration of a foreign body in the forearm. Ultrasonographic localisation and surgical removal of the foreign body resulted in gradual recovery of neurological function. PMID- 11478067 TI - Mycobacterium chelonae lumbar spinal infection. AB - A case report of a previously healthy adult patient with a lumbar spinal extradural abscess due to Mycobacterium chelonae is presented. His course of treatment was complicated by recurrent psoas abscesses, as well as multiantibiotic resistance, requiring multiple surgical drainage procedures and antibiotic changes over a 33-month period. Cure was achieved only after aggressive surgical debridement of the abscess. PMID- 11478068 TI - Delayed aneurysm rerupture following total endovascular occlusion. AB - Endovascular coiling is a well established technique for the treatment of selected intracranial aneurysms, but its long-term efficacy, including the rate of rehaemorrhage from treated lesions, remains to be clearly determined. We report a case in which the rerupture of a small aneurysm occurred 12 months after embolization, despite angiographic occlusion on immediate post-procedural and 6 month check angiography. PMID- 11478069 TI - Dural arteriovenous malformation associated with recurrent subdural haematoma and intracranial hypertension. AB - An unusual case of intracranial hypertension and symptoms of a left parieto occipital mass lesion due to a dural arteriovenous malformation (AVM) with a large and dilated draining vein is reported. The patient also had a history of homolateral recurrent subdural haematoma, 11 years before. Subdural haematoma is rarely associated to a dural AVM. We suggest that the recurrent subdural haematoma was due to the very slow and intermittent venous bleeding from the preexisting dural malformation, which progressively enlarged in the following years to become very large. The symptoms of intracranial hypertension and papilloedema may be explained by the increased pressure in the dural sinus and the cerebral venous system. On the other hand, focal neurological symptoms in our case resulted from the mass effect due to an aneurysmally dilated draining vein in the left parieto-occipital region. PMID- 11478070 TI - Ependymoma of the neurohypophysis. AB - There are two reported cases of ependymomas arising in the pituitary fossa; one in a human, the other in a horse. Both died during their stay in hospital. The case presented here is the first published case of a patient who is well 3 months after surgery. PMID- 11478071 TI - Chiari type I malformation in two monozygotic twins. PMID- 11478072 TI - New models for analysing hydrocephalus and disorders of CSF volume transmission. PMID- 11478073 TI - Neurocysticercosis in pregnancy. PMID- 11478074 TI - Rational use of antibiotics in the treatment of brain abscess. PMID- 11478075 TI - Twist-drill skull placement of bolts and drains. PMID- 11478076 TI - Endoscopic fenestration of a symptomatic cyst of the septum pellucidum. PMID- 11478077 TI - Treated brain angioma patients and licence to drive. PMID- 11478078 TI - OIG, HCFA and HIPAA legalities explained. PMID- 11478079 TI - The realities of HIPAA and how to incorporate them into your practice. PMID- 11478080 TI - A snapshot of 21st century telemedicine in Michigan. PMID- 11478081 TI - Quality and speed of patient care improve with telemedicine technology. PMID- 11478082 TI - The days of steady PL premiums are numbered. PMID- 11478083 TI - The changing role of physicians in home health care. PMID- 11478084 TI - Bioterrorism be prepared for this real threat. PMID- 11478085 TI - End-of-life guide. Offers a comprehensive look at care. PMID- 11478086 TI - MSMS alliance focuses on violence prevention. PMID- 11478087 TI - National practitioner data bank reporting requirements. PMID- 11478088 TI - Respecting what we destroy. Reflections on human embryo research. PMID- 11478089 TI - Strap him down. PMID- 11478090 TI - Gang aft agley. PMID- 11478091 TI - The gift of life and the common good. The need for a communal approach to organ procurement. PMID- 11478092 TI - Tube feeding and advanced progressive dementia. PMID- 11478093 TI - Phase I cancer trials: therapeutic research? PMID- 11478094 TI - The Maltese conjoined twins. Two views of their separation. PMID- 11478095 TI - The Maltese conjoined twins. A separate peace. PMID- 11478096 TI - Phase I cancer trials: therapeutic research? PMID- 11478097 TI - The Maltese conjoined twins. Hubris in the court. PMID- 11478098 TI - Let us imagine.... PMID- 11478099 TI - Hand transplant recipient throws in the towel. PMID- 11478100 TI - The point of a ban. Or, how to think about stem cell research. PMID- 11478101 TI - The business ethics of bioethics consulting. PMID- 11478102 TI - Seeing the duties to all. PMID- 11478103 TI - Doing good and doing well. PMID- 11478104 TI - Spiritual care at the end of life. PMID- 11478105 TI - An extremely urgent transplantation? PMID- 11478106 TI - The human right to health: a right to the "highest attainable standard of health". PMID- 11478107 TI - Babel, justice, and democracy: reflections on a shortage of interpreters at a public hospital. PMID- 11478108 TI - Decisionmaking issues in the rehabilitation process. PMID- 11478109 TI - Prenatal genetic testing. PMID- 11478110 TI - The ethics and economics of heroic surgery. PMID- 11478111 TI - Revisiting the Belmont report. PMID- 11478112 TI - Postmodern illness. PMID- 11478113 TI - The BMA addresses Britain's rationing problem at last. PMID- 11478114 TI - AMA's E-force enters patient privacy debate. PMID- 11478115 TI - Gene-poor vermin: humanity's new place in nature. PMID- 11478117 TI - Throwing a bone to the watchdog. PMID- 11478116 TI - No end in sight for final rules on medical privacy. PMID- 11478118 TI - The ethics wars. Disputes over international research. AB - The effort to revise the Declaration of Helsinki and the CIOMS Guidelines has sparked a sometimes vitriolic debate centering on the use of placebo controls. PMID- 11478119 TI - Swinging on the pendulum. Shifting views of justice in human subjects research. AB - Federal policies on human subjects research have performed a near-about face. In the 1970s, policies were motivated chiefly by a belief that subjects needed protection from the harms and risks of research. Now the driving concern is that patients, and the populations they represent, need access to the benefits of research. PMID- 11478120 TI - Arizona's cancer clinical trials law: flawed process, flawed product. PMID- 11478121 TI - Pain and sickle cella anemia. PMID- 11478122 TI - Defining research misconduct: will we know it when we see it? PMID- 11478123 TI - Reading between the lines: direct-to-consumer advertising of genetic testing. PMID- 11478124 TI - Literature, literary studies, and medical ethics: the interdisciplinary question. AB - How do we know what is right, or before that, how do we recognize what is morally salient? Such matters lie deeper than can be plumbed by traditional philosophical modes of inquiry alone. Careful study of them requires also the study of literature, with the meticulous appraisal that it encourages of the intricate, tangled issues involved in apprehending the world, finding our way in it, and representing it to others. In this way, the study of literature contributes to a richer and more complex perspective on moral problems, and a more cautious view of the status and breadth of attempts to solve those problems. PMID- 11478125 TI - Physician-assisted death for the terminally ill. PMID- 11478126 TI - The monopoly stage of capitalism. PMID- 11478127 TI - Goodbye to all that. The end of moderate protectionism in human subjects research. AB - Federal policies on human subjects research have undergone a progressive transformation. In the early decades of the twentieth century, federal policies largely relied on the discretion of investigators to decide when and how to conduct research. This approach gradually gave way to policies that augmented investigator discretion with externally imposed protections. We may now be entering an era of even more stringent external protections. Whether the new policies effectively absolve investigators of personal responsibility for conducting ethical research, and whether it is wise to do so, remains to be seen. PMID- 11478128 TI - Utilization of ambulatory medical care by women: United States, 1997-98. AB - OBJECTIVES: This report presents national estimates of the volume and characteristics of ambulatory medical care provided to women 15 years of age and over in the United States. Included is information on the characteristics of the patients, providers, and visits. A section on comparative differences in use by sex is also included. SOURCE OF DATA: This report is based on an analysis of data from the 1997 and 1998 National Ambulatory Medical Care Survey (NAMCS) and National Hospital Ambulatory Medical Care Survey (NHAMCS), national probability sample surveys of visits to office-based physicians (NAMCS) and visits to the outpatient departments and emergency departments of non-Federal, short-stay and general hospitals (NHAMCS) in the United States. Sample data are weighted to produce annual estimates. RESULTS: Approximately 500 million visits were made by women to ambulatory medical care providers annually in 1997 and 1998, representing an age-adjusted rate of 4.6 visits per woman per year. The rate of ambulatory medical care visits increased with age (3.8 per woman 15-44 years of age, 4.7 visits per woman 45-64 years of age, and 7.1 visits per woman 65 years of age and over). Six out of ten visits to office-based physicians and hospital outpatient departments had no mention of therapeutic or preventive services provided. Nonnarcotic analgesics, antidepressant, and estrogen/progestin were the three most common classifications of medications mentioned. Compared with visits by men, visits by women (with nonpregnancy-related diagnoses) were more frequent at younger ages and more likely to be to primary care physicians and outpatient departments. PMID- 11478129 TI - [Two elderly patients with advanced gastric cancer responding to chronomodulation chemotherapy with tegafur + cisplatin + isovorin followed by oral administration of S-1]. AB - Recently, as society ages there have become more elderly gastric cancer patients with/without several complications(cerebrovascular diseases, cardiac diseases, atherosclerosis, DM, etc.), that were non-resected and require highly effective chemotherapy and good QOL. We report two elderly gastric cancer patients responding to chronomodulation chemotherapy (tegafur + cisplatin + Isovorin) based on circadian rhythms plus a new antitumor drug, S-1. The treatment protocol was tegafur 800 mg/body, days 1-7 (continuing 16 h, intravenously with 500 mg/body from 16 to 0 h, 300 mg/body from 0-8 h, for non-uniform administration), cisplatin 10 mg/body, days 1-5, (16 h), Isovorin 25 mg/body, days 1-5, (16 h, oneshot infusion, for 4 courses followed by a week rest. Next was S-1 120 mg/body x 2 times orally for 28 days, followed by 2 weeks rest, the administered for another 28 days. The first patient was 74 years of age, with advanced type 3 plus early type IIc gastric cancers with liver metastasis (H1). After chemotherapy the liver metastasis disappeared, there was a 70% reduction in the advanced cancer and the early cancer disappeared. The second patient was 84 years of age, with advanced type 3 gastric cancer invading the esophagus. After chemotherapy, the primary lesion was reduced 80% and the esophageal invasion mass shrunk. The only adverse effect was grade 2 pancytopenia. In conclusion this regimen resulted in good intrachemotherapeutic QOL and highly effective performance in elderly advanced gastric cancer patient. PMID- 11478130 TI - [Intermittent administration of 5-FU and isovorin to patients with advanced and recurrent colon cancer]. AB - We attempted a new regimen of intermittent administration of 5-FU and low-dose Isovorin (F.I) to four patients with advanced and recurrent colon cancer. A partial response (PR) was achieved in two of four patients who had evaluable lesions for this treatment. We observed few side effects among these patients. Only one patient among four showed grade 1 neuropathy after two administrations of this chemotherapy. However, after a two-week pause in administration, the neuropathy disappeared and we could continue the therapy. This patient with multiple liver metastases achieved a partial response. Other patients had no side effects such as bone marrow suppression or digestive symptoms. This intermittent F.I treatment might be an effective and promising therapy with few side effects even for patients with poorer conditions. PMID- 11478131 TI - [Usefulness of chemotherapy with CPT-11 in clinic for three colorectal cancer in terminal stage]. AB - In our clinic we performed chemotherapy with CPT-11 in three cases of non resectable advanced colorectal cancer, including two cases of multiple liver metastasis and one case of multiple lung metastasis, and obtained various alleviating effects. Heretofore two factors. 1. direct effect and 2. longevity effects, have been focused on when evaluating the effect of various chemotherapy regimens. However, the symptoms of these patients were alleviated by the chemotherapy without obtaining either of the two effects. Furthermore, their performance statuses were improved. It is therefore sufficiently beneficial for patients in the terminal stage to undergo the chemotherapy. The patient in the terminal stage should be taken care of in the clinic rather than in the hospital. At present, safe and effective therapies on an ambulant basis have not been established. It will now be necessary for us to evaluate the many accumulated cases. It is conceivable that chemotherapy with CPT-11 in the clinic would be extremely useful for patients in the terminal stage for the purpose of improving the QOL, if the dose and administration interval of CPT-11 are given sufficient attention. PMID- 11478132 TI - [Feasibility, activity, and change in the level of blood paclitaxel concentration after weekly paclitaxel therapy for a patient with recurrent ovarian cancer]. AB - The patient was a 59-year-old woman with recurrent ovarian cancer. A CT scan of the abdomen showed enlargement of abdominal para-aortic lymph nodes (PAN) after the primary operation and 8 cycles of the combination chemotherapy with paclitaxel (TXL) and carboplatinum (CBDCA). As a second line chemotherapy for the patient, weekly administration of TXL (60 mg/m2/week x 3 weeks) was given. The toxicity was acceptable and less pronounced than with the standard TXL + CBDCA therapy. Peak blood TXL concentration, about 90 ng/ml, was achieved 4 hours after the administration of TXL. The blood TXL concentration was below the detectable limit 48 h after the administration of TXL. An almost 50% shrinkage in the size of the PAN was obtained after 2 cycles of treatment. Good QOL is being maintained without any repeated aggravation of the tumor. PMID- 11478133 TI - [A preliminary report on the treatment of pleural carcinomatosis with SMANCS]. AB - To clarify the effect of SMANCS on malignant pleural carcinomatosis, seven patients with malignant pleural effusion were treated with SMANCS administered via an intracavitary route. Five patients showed improvement after one or two injections of SMANCS into the thoracic cavity, although 2 patients needed further therapy with the immunopotentiating agent picibanil (OK-432). No serious adverse effects were observed. This simple therapeutic tactic with SMANCS may be effective in cases of malignant pleural carcinomatosis. PMID- 11478134 TI - [Targetting intra-arterial infusion chemotherapy with nedaplatin for oral cancer]. PMID- 11478135 TI - [Tumor markers in breast cancer]. AB - Many serological markers have been utilized to indicate the status, risk, or presence of breast cancer. In May 1996, the American Society of Clinical Oncology (ASCO) convened a Tumor Marker Panel and determined clinical practice guidelines for the use of tumor markers in breast cancer. Eight markers containing carcinoembryonic antigen (CEA) and CA15-3 were evaluated and assigned by expert reviewers to be valuable markers of breast cancer. CA15-3 recognizes a mucin-like glycoprotein, MUC-1, which is frequently expressed in breast cancer tissues. BCA225, which may recognize antigens similar to MUC-1 glycoprotein, are sensitive and specific markers for breast cancer. However, it is not recommended to measure the 2 markers in combination. The measurement of carboxy-terminal telopeptide of type I collagen (I CTP) is worthwhile as a serological diagnostic method of bone metastasis from breast cancer. Other markers such as erbB-2, CYFRA 21-1 and PTHrP are candidates for clinical utilization as tumor markers in breast cancer. PMID- 11478136 TI - [Molecular and cellular biological analysis on cancer cachexia syndrome]. AB - About half of patients with cancer will suffer from wasting syndrome, called cancer cachexia, which shows abnormality of homeostasis, nutrition, endocrine function, metabolism, immunity et al. This syndrome is characterized with anorexia and weight loss caused by degradation of skeletal muscle and adipose tissue. Progressive weight loss is responsible not only for a poor quality of life and poor response to anti-cancer drug, but also shorter survival time comparing patient without weight loss. Various factors have been found as mediators of this syndrome base on the development of immunology, biochemistry and molecular and cellular biology. These include several cytokines, proteolysis inducing factor (PIF), lipid-mobilizing factor (LMF), apoptosis-inducing factor and another factors. Recentry, molecular biological analysis makes clear more detail mechanisms of cancer cachexia syndrome, for example, ubiqutin/proteasome pathway, activation of nuclear transcriptional factors and others. These progresses will contribute not only to establish new treatment but also to carry out "order-made palliative oncology" using DNA-chip and/or Protein-chip in near future. PMID- 11478137 TI - [Developments of hormonal agents for breast cancer]. AB - Endocrine therapy for breast cancer, which began with ovariectomy, has a history of more than 100 years. Tamoxifen has been an epoch-making drug during the late 20th century. Recently LHRH analogues which work as downregulators of estrogen production and many aromatase inhibitors (AI), both non-steroidal and steroidal have been developed in Japan. Fadrozole was the only AI until anastrozole was approved a few months ago. Letrozole was shown to be a better AI than fadrozole by prospective randomized double blind examination; however, it is not licenced yet. The reason is that the appropriate dosage is not identical to that of Western countries. We have reached a time when order-made medicine should take into consideration such differences as race and individuality. PMID- 11478138 TI - [Current status of adjuvant endocrine therapy for hormone responsive breast cancer]. AB - Three important meetings on adjuvant hormone therapy for breast cancer were held recently: the 5th EBCTCG Meeting, NIH Consensus Meeting, 7th International Conference on Adjuvant Therapy of Primary Breast Cancer. The conclusions of these meetings are: 1. adjuvant hormone therapy should be indicated only for patients with estrogen/progesterone receptor positive cancer, 2. five years of tamoxifen is the standard care at present, 3. ovarian ablation by any means has been proved effective in premenopausal patients and LH-RH agonist should be given at least two years, and 4. aromatase inhibitors should not be used in clinical practice, because several prospective randomized trials are ongoing at present. The patients treated with LH-RH agonist combined with tamoxifen showed better relapse free survival compared with LH-RH agonist alone in the INT-0101 trial. This was an important trial because combined hormone therapy had not been proven more effective than individual hormone therapy previously. Combined hormone therapy including LH-RH agonist may be considered in premenopausal patients. There is a growing consensus that chemotherapy is effective through the ovarian suppression. In this sense, hormonal therapy should be considered first for hormone responsive patients. On the contrary, standard chemotherapy has shifted from CMF combination to an anthracycline containing regimen. Chemoendocrine therapy may be considered in high risk patients. PMID- 11478139 TI - [Endocrine therapy for advanced or recurrent breast cancer]. AB - Endocrine therapy of advanced or recurrent breast cancer was described. The presence of ER or PgR in primary breast tumors is the best-established marker for response to endocrine therapy. However, ER-positive breast tumors overexpressing EGF-R and/or HER-2 (Erb B2) are resistant to endocrine therapy. Recently it was suggested that an elevated level of the circulating extracellular domain of HER-2 could be a predictor for poor response to endocrine therapy. LH-RH agonist is used as a first-line therapy for premenopausal patients. And LH-RH agonist plus tamoxifen (TAM) has shown a higher response rate and more prolonged survival than LH-RH agonist or TAM alone. As two new aromatase inhibitors, anastrozole (ANA) and letrozole, have shown an equal or higher response rate and a prolonged time to progression than TAM as a first-line therapy, these could be used as a first line therapy instead of TAM. In a cross-over trial of ANA and TAM, the response rate of ANA after TAM failure was equal to that of TAM after ANA failure. As these drugs showed an equal or higher response rate and longer survival than progestin in TAM resistant cases, these drugs could also used as a second-line therapy. In addition, the trend of recent studies regarding the mechanisms of hormone resistance is also described. PMID- 11478140 TI - [Management of a hormone dependent cancer with endocrine therapy--prostate cancer]. AB - We offer a historical overview of endocrine therapy for prostate cancer. Hormone therapy remains the cornerstone of treatment for patients with locally advanced or metastatic prostate cancer. Although this therapy has been traditionally performed by oral estrogen or bilateral orchiectomy, there are now two most important pharmacological hormonal therapies: LH-RH agonist and antiandrogen therapy. We do not have yet sufficient data to conclude whether maximal androgen blockade from the combined use of an LH-RH agonist and an antiandrogen will prolong the survival in patients with metastatic prostate cancer, nor to conclude whether neoadjuvant androgen ablation therapy improves the disease-free survival of patients after radical prostatectomy. New treatment strategies and modalities such as LH-RH antagonists, intermittent hormonal therapy, and antiandrogen monotherapy are appearing and being tested in clinical trials. However, to date there is still no effective therapy for patients who have hormone refractory disease. PMID- 11478141 TI - [Neoadjuvant hormonal therapy for prostate cancer]. AB - There have been expectations that neoadjuvant hormonal therapy would decrease the rate of positive surgical margins and, therefore, to improve the patient's survival rate after radical prostatectomy for clinically localized prostate cancer. A review of seven prospective randomized studies for clinically localized prostate cancer revealed a significant decrease in the positive surgical margin rate in cases of clinical T2 disease after neoadjuvant hormonal therapy with prostatectomy. However, this treatment did not alter the rate of seminal vesicle invasion or lymph node metastasis after radical prostatectomy. There was no difference in operative blood loss, operating time, complication rate or hospital stay between patients treated with neoadjuvant hormonal therapy and controls. Furthermore, there was no improvement in prostate specific antigen-free survival rate after a maximum of 4 years follow-up. Further research is required to determine the optimal duration of neoadjuvant hormonal therapy and whether this therapy increases the survival rate. Neoadjuvant hormonal therapy before radical prostatectomy is not supported by any outcome at this point and its use remains in the investigational stage. PMID- 11478142 TI - [Hormonal therapy for endometrial adenocarcinoma]. AB - Recently, the number of cases of endometrial cancer has increased in Japan. Most of the increase are accounted for by premenopausal cases, which are frequently positive for ER or PR. Hormonal treatment using progestins such as MPA has been widely applied to endometrial cancer patients under 40 years old under the conditions of grade 1 well-differentiated endometrioid adenocarcinoma without myometrial invasion. In our hospital, we applied high-dose (600 mg/day) MPA for over 8 weeks in 14 cases of endometrial cancer, of which adenocarcinoma disappeared in 12 cases (86%), followed by cyclic administrations of low-dose MPA, with 7 subsequent recurrences. We think that a protocol for improving ovarian function, such as active induction of ovulation, should be established to induce intrinsic progesterone sufficient for the prevention of the recurrence of endometrial cancers. In the 2 cases, in which adenocarcinoma remained even after long administrations of MPA, myometrial invasion was noted in the surgically resected specimens. For advanced or recurrent endometrial cancers, MPA has been reported to be effective in an average of 26% in several reports, and effective in 42% cases when applied with combination chemotherapy, such as CAP, by virtue of the "chemical modulator" effect, which can delay the acquired resistance against ADM or CDDP. Furthermore, MPA has resulted in a significant improvement of 5-year disease-free survival rate when used as adjuvant therapy after complete operations and whole pelvic irradiation, compared with administrations of 5-FU in a randomized controlled study in Japan. Thus, in the future, we consider that hormonal therapy will play a more important role in endometrial cancer treatment. PMID- 11478143 TI - [Combination chemotherapy of carboplatin and docetaxel for advanced non-small cell lung cancer (NSCLC)]. AB - Since mid-1997, we have treated 60 patients with advanced NSCLC with carboplatin (CBDCA) plus docetaxel (TXT). CBDCA (300-400 mg/m2) and TXT (60 mg/m2) were given on day 1 every 3 weeks. The mean treatment cycle was 2.3 +/- 1.4 (range 1-7). Fifty-four patients had measurable tumors, of whom 2 patients achieved a complete response and 19 patients achieved a partial response (37.0%, 95% CI: 24.3-51.3%) (Ad 12/29, Sq 7/23, Lar 1/2). Median survival time was 12.8 months and 1 year survival was 53.6%. AUC of CBDCA was not related to response (AUC of responders and non-responders was 3.29). Myelosuppression was moderate (WBC 2,284 mm3, range 800-4,700, PLT 16.4 x 10(4) mm3, range 6.6-41.5 x 10(4), Hb 10.9 g/dl, range 6.4 15.8). Leukocytopenia was related to AUC of CBDCA (R2 = 0.1093) but thrombocytopenia was not related to AUC of CBDCA (R2 = 0.0553). Gastrointestinal toxicity was mild (grade 0-1: 57%, grade 2: 35%, grade 3: 8%, grade 4: 0%). Treatment with CBDCA plus TXT combination is safe and effective in patients with NSCLC. PMID- 11478144 TI - [Examination on efficacy and safety of concurrent use of ondansetron hydrochloride and steroid in lung cancer patients on cisplatin]. AB - The anti-emetic effect and safety in patients receiving ondansetron hydrochloride (OND group) and concurrent use of ondansetron and dexamethasone (DEX group) in cases of acute and delayed onset emesis induced by a single high dose of cisplatin, given as a chemotherapy to lung cancer patients, were comparatively studied. The study subjects were 78 lung cancer patients. The OND group received 4 mg of ondansetron via slow intravenous injection on Day 1, 30 minutes prior to cisplatin, and for Days 2 to 5, the subjects orally received 4 mg ondansetron tablet each day. The DEX group received the same dose regimen of ondansetron as the OND group for Days 1-5, but in addition the subjects received dexamethasone injection in doses of 8 mg twice daily on Day 1 and 4 mg (1 mg QID) daily for Days 2-5. An anti-emetic effect against acute nausea and vomiting was achieved in 83.8% of the OND group and in a higher rate of 94.6% of the DEX group. Significantly better efficacy was seen in the DEX group as to the complete suppression rate of nausea and vomiting and the improvement of food intake. The group also achieved better efficacy in delayed onset of emesis. Two cases of adverse reactions (hiccups and elevation of ALT and BUN) were observed in the DEX group; however, since the symptoms were all mild, we did not consider there was any problem in safety. We conclude from the above findings that concurrent administration of ondansetron hydrochloride and dexamethasone is a clinically useful treatment for acute and delayed onset emesis induced by a single high dose of cisplatin given to lung cancer patients. PMID- 11478145 TI - [Clinical results of concurrent chemoradiation therapy for primary highly advanced esophageal cancer]. AB - Clinical results after non-curative resection of highly advanced esophageal cancer are extremely poor. We administered concurrent chemoradiation therapy (CRT) as a multidisciplinary therapy in cases of highly advanced esophageal cancer for which non-curative resection is expected. The efficacy rate of the therapy was 59.4%, and the 3-year-survival rate 10.2%. A life-prolonging effect (the 3-year survival being 17.9%) was observed in the effective cases. Our future aim is to establish a safer, more reliable and cost-effective therapy by estimating the degree of efficacy before administration of CRT and then selecting cases suitable for CRT. PMID- 11478146 TI - [A phase I study of docetaxel (TXT) and doxifluridine (5'-DFUR) combination therapy in patients with advanced and recurrent breast cancer]. AB - We investigated efficacy and tolerance of chemotherapy with doxifluridine (5' DFUR) and docetaxel (TXT) in advanced/recurrent breast cancer. Subjects were enrolled by central registration. The regimen included 5'-DFUR orally for 14 consecutive days, and TXT intravenously on day 8. It was repeated every 3 weeks, as long as possible, including dosage levels of 5 scheduled steps. Patient registration was started in August 1999 and 5 patients given level 1 regimen (5' DFUR, 800 mg/day; TXT, 50 mg/m2) were evaluated. Although the results revealed neutropenia of grade 3 in 4/5 patients and leukocytopenia in 2/5 patients, no other side effects were observed. Taking into consideration the toxicity profiles of each drug in level 1, a level 2b regimen (5'-DFUR, 800 mg/day; TXT, 60 mg/m2) was accepted. Seven patients were registered for level 2b dosage and were examined for the safety of the regimen. Two patients discontinued the level 2b regimen due to percutaneous adverse reactions (DLT) and 6/7 patients developed neutropenia of grade 4. Clinical effects in level 1 group included: 1 CR, 2 PR, 1 long NC (NC longer than 24 weeks), and 1 NE, for a response rate of 60.0% (3/5 patients). Those in level 2b included: 2 CR, 2 PR, 1 NC, and 2 NE, for a response rate of 57.1% (4/7 patients). Based on the safety and efficacy of the combined therapy, the recommended dosage of this regimen is 5'-DFUR, 800 mg/day, combined with TXT, 60 mg/m2. A Phase II study is being conducted using this dosage. PMID- 11478147 TI - [The efficacy of combination chemotherapy of 5'-deoxy-5-fluorouridine (5'-DFUR), cyclophosphamide (CPA) and medroxyprogesterone acetate (MPA) for bone metastasis in breast cancer patients]. AB - 5'-DFUR is a pro drug of 5-FU, which is known to be converted by thymidine phosphorylase (dThdPase). A recent pre-clinical study revealed that CPA upregulates dThdPase activity specifically in tumor cells. Furthermore, clinical trials have shown significant response rates in breast cancer patients, when using the chemotherapy combination of 5'-DFUR, CPA and MPA. The purpose of this study was to examine the efficacy of this regimen as a pain reduction therapy for breast cancer patients with bone metastasis. Ten patients who had bone metastasis with restricted ADL were included in the study. All of the patients had had previous exposure to such standard chemotherapy as CAF, CMF, taxol and oral 5-FU administration. The patients were administered daily oral doses of 5'-DFUR at 800 1,200 mg, CPA at 200 mg and MPA at 400-800 mg for two weeks as induction therapy, followed by two weeks rest (one to two cycles). Daily dose of 800 mg of 5'-DFUR, 100 mg of CPA, 400-800 mg of MPA was continuously administered thereafter. The main findings included a significant decrease in pain in eight patients, which continued for more than 6 months. In five patients, the effect lasted more than one year. As the pain decreased, the patients' QOL was improved. Hematological toxicity of more than grade 3 was observed in three patients but only during the induction therapy. One patient had pulmonary thrombosis and required hospitalization. In conclusion, oral administration of 5'-DFUR/CPA/MPA is well tolerated and useful in reducing pain. PMID- 11478148 TI - [Concurrent chemoradiation therapy with nedaplatin for high-risk cervical cancer- clinical investigation of adverse events]. AB - A clinical investigation of adverse events was conducted to confirm the safety of concurrent chemotherapy using nedaplatin (cisplatin derivative) and radiotherapy in the high-risk carcinoma of the uterine cervix. Seven patients who were treated with radical radiotherapy and 5 patients who were treated with adjunctive radiotherapy after radical hysterectomy and pelvic lymphadenectomy were eligible for the study. Nedaplatin was given intravenously at 70 mg/m2 on day 1 and day 29, and a total of 24 courses of nedaplatin administration were observed. None of the planned radiotherapy was postponed or discontinued due to side effects. Major adverse effects observed were gastrointestinal effects such as anorexia (66.7%), nausea and vomiting (33.3%) and diarrhea (66.7%). Grade 3 (in the 2nd course) and Grade 4 (in the 1st course) diarrhea was observed in one patient, which was easily relieved by antidiarrheal. Hematologic side effects were also major, including leukopenia (62.5%), neutropenia (75.0%), anemia (75.0%), and thrombocytopenia (33.3%). Hematologic effects were generally moderate; no Grade 4 (severe) effects were observed. Although these hematologic effects were lasting longer compared with radiation therapy alone, there were no significant differences in the seriousness of these side effects. Concurrent chemoradiation therapy with nedaplatin 70 mg/m2 every 4 weeks was safe and adverse effects were self-limited or resolved with medical management. Dose escalation in the phase III clinical study may be considered. PMID- 11478149 TI - [A case of stage IV advanced lung adenocarcinoma treated effectively using paclitaxel with the fibrosing radiographically on the chest CT]. AB - A 48-year-old man presented with dyspnea. He was diagnosed as having advanced lung adenocarcinoma of stage IV (T4N2M1). After 3 courses of paclitaxel therapy every 3 weeks intravenously, subjective symptoms, arterial blood gas and radiographical findings improved very much. In the process of the tumor reduction, chest CT showed unique radiographical findings of so-called lung fibrosis over the entire lung and reduction of lung volume. An additional 3 courses of weekly paclitaxel therapy have been given on an outpatient basis. After the change of therapy, the reduction in the tumor has continued and the tumor progression is under control without a decline in the patient's QOL from the start of the chemotherapy over 6 months to the present. PMID- 11478150 TI - [A case of advanced gastric cancer with multiple liver metastases in an elderly patient that responded dramatically to UFT therapy]. AB - An 88-year-old female patient suffering from Borrmann type 3 advanced gastric cancer complicated by multiple hepatic metastases underwent a total gastrectomy. A small dose of 200 mg/day of UFT was administered orally every day postoperatively. At postoperative month 6, a marked diminishment of the hepatic metastatic lesions was noted. Resection of the primary lesion with a regimen of a small oral dose of UFT was remarkably effective in this elderly gastric cancer patient with complications from multiple hepatic metastases. PMID- 11478151 TI - [A case of advanced gastric cancer that was resectable after asctic fluid had disappeared following administration of TS-1]. AB - A 60-year-old man who had suffered from epigastic pain and general malaise from November 1999 was admitted to our hospital due to Borrmann type 3 gastric cancer with ascites on December 7, 1999. We considered a radical B operation impossible, and placed the patient on neoadjuvant TS-1 chemotherapy consisting of 1 M tegafur, 0.4 M gimestat, and 1 M otastat potassium. There were no side effects other than Grade 1 nausea and mild loss of appetite throughout the chemotherapy. After 8 weeks of administration, the primary lesion was reduced in size, and ascitic fluid had disappeared on abdominal computed tomography images. Therefore, a total gastrectomy with splenectomy and D2 lymph node dissection was performed on March 31, 2000. This was a radical B operation that was not possible earlier. The pathological diagnosis was tub2, SE, N1, CY0, H0, P0, M0, INF gamma, ly1, v1, PM (-), DM (-) and the antitumor efficacy of TS-1 was Grade 2 histologically. The patient remains alive and in good condition with no relapse of the gastric cancer 8 months after surgery. PMID- 11478152 TI - T cell receptor and function cosegregate in gamma-delta T cell subsets. PMID- 11478153 TI - Role of the IL-7 receptor in gamma-delta T cell development. PMID- 11478154 TI - Epidermal and dermal gamma-delta T cells. PMID- 11478155 TI - Cytokine profiles and responsiveness of dendritic epidermal T cells: cytokine mediated communication with other cells. PMID- 11478156 TI - Gamma-delta T cells with emphasis on their functional role in the epidermis. PMID- 11478157 TI - Role of gamma-delta T cells in cutaneous hypersensitivity reactions. PMID- 11478158 TI - Gamma-delta T cells: immunoregulatory functions and immunoprotection. PMID- 11478159 TI - [Functional consequences of cleft palate and its management]. AB - On the basis of the experience with 5200 various types of orofacial cleft operations performed during 42 years (1959-2000) and of the 60-70 cases with velopharyngeal insufficiency without cleft examined yearly author stresses that the surgical closure of the cleft is not enough in the care: the functional consequences (respiratory, sucking, swallowing, speech, hearing and maxillofacial developmental disorders) should also be managed. The first symptom of Robin sequent (cleft palate associated with dysgnathia) is respiratory disorder improving mostly spontaneously but worsening sometimes after palatoplasty. Polysomnography has been performed by the author routinely before all primary palatoplasties for establishing the possible surgical risks and for choice of the optimal time for surgery. On the basis of the results in 61 infants, the surgery was contraindicated and postponed in 6 cases. The various orofacial clefts may cause swallowing problems in different phase of swallowing and different types of dysphagia: the cleft lip cause sucking problems in the preparatory phase, the cleft palate in the oral transitory phase and the velopharyngeal insufficiency in the pharyngeal phase, but the sensomotor function is more decisive in the swallowing process than the cleft itself. Use of an obturator is not necessary, long-term catheter feeding is inappropriate, early closure of the soft palate (in the age of 8-10 months) is recommended. The liquid or food reflux through the nose can be established by cinefluoroscopy with contrast material and with nasoendoscopy following the way of coloured boluses. Retarded speech development, hyperrhinophony, nasal escape, facial grimacing, articulation disorders and dysphonia are the most frequent voice and speech disorders; speech therapy is in 70%, velopharyngoplasty in 20% of the cases indicated. Anatomical result was good in 98% of 1107 flap surgeries operated on by the author, hyperrhinophony ceased in 90%. The results are assessed by a 5-grade perceptual scale. Among the instrumental procedures videofluoroscopy, videoendoscopy, and nasometry seems to be the most informative. The cause of the frequent hearing disorders is mostly the eustachian tube dysfunction. Author found pathological tympanograms in 64% of their cases. The hearing slightly improves after staphylorraphy and often after flap surgery. Adenoidectomy and grommet insertion are alternative procedures in the therapy. The surgeries have an effect on the maxillofacial growth but this is not significant. Multidisciplinary co-operation in the management of cleft patients is indispensable. PMID- 11478160 TI - [Prevalence of human papillomavirus infections in our five-year data]. AB - Human papillomavirus infection proved to be the most important risk factor for the development of cervical cancer and its preblastomatosis. Human papillomavirus was detected from 1996. June to 2000. September at 1635 patients, who had been positive by colposcopy and/or cytology in an earlier examination. The place of the study were our outpatients' departments and consultations by specialists of Debrecen University, Department of Obstetrics and Gynecology. Hybrid capture system was used to demonstrate the presence of the virus and managed to prove it in the 28.9% of cases. 3.1% of the patients (51 persons) had acquired low-risk, and 23.6% (386 persons) high risk virus types, however 2.1% of the woman (35 patients) were infected with both low-risk and high-risk human papillomavirus types at the same time. Long time decrease of virus prevalence was observed after the age of 35 year, and the significant degree and timing decrease of after the age of 30 year at patients infected with combination of low-risk and high-risk virus types, respectively. This observation is indicative of the correlation between colposcopic-, cytologic abnormalities and the persisting high-risk human papillomavirus infections. PMID- 11478161 TI - [Successful new method for surgery of the postinfarctional left ventricular free wall rupture]. AB - Authors discuss a history of a 78-years old male who suffered from left ventricular free wall rupture due to acute transmural myocardial infarction. Cardiac surgical intervention was carried out immediately after the ECHO examination which established the diagnosis. First time in Hungary authors used a sutureless technique to close the rupture and utilised double component glue and synthetic patch for this purpose. The patient had uneventful recovery and remained free of symptoms throughout his three months period of follow up. Current available data and knowledge are summarised about this life threatening disease. PMID- 11478162 TI - [Endoscopic removal of a toothbrush from the esophagus using a polypectomy device]. AB - A rare case is reported with a large foreign body in the upper gastrointestinal tract. The 32-year-old man accidentally swallowed his toothbrush which was successfully removed via endoscopy using polypectomy snare under local anesthesia. PMID- 11478163 TI - [Dr.Jozsef Imre, Sr. - his professional work with special emphasis on insurance medicine]. PMID- 11478164 TI - [Memory of Karoly Lechner (1850-1922)]. PMID- 11478165 TI - [Evolution and state of the art of extracorporeal perfusion in Hungary]. PMID- 11478166 TI - "Discharge patients sooner, or we'll discharge you". PMID- 11478167 TI - Finally earning "real money"--now what? PMID- 11478168 TI - These doctors cope with disability. Could you? PMID- 11478169 TI - Why premiums are soaring again. A new "malpractice crisis"? PMID- 11478170 TI - Where a war on doctors has already begun. PMID- 11478171 TI - Take a hard line on software support. PMID- 11478172 TI - Your obligations when doing pre-employment exams. PMID- 11478173 TI - Rx price controls: Maine gets a go-ahead. PMID- 11478174 TI - Claims in, approvals out--in seconds. PMID- 11478175 TI - School shootings: doctors were victims, too. PMID- 11478176 TI - Maternity leave policies that really deliver. PMID- 11478177 TI - Court to FP: the OR is off limits! PMID- 11478178 TI - Who'll lead the way to a better system? Panel discussion. PMID- 11478179 TI - Seismic shift in Washington. PMID- 11478180 TI - Senate HELP committee holds hearing on patient safety. PMID- 11478181 TI - Senate Special Committee on Aging holds hearing on technology and prescription drug safety. PMID- 11478182 TI - Congressional healthcare outlook: prescription drugs. PMID- 11478183 TI - Members of Congress continue to voice concern over privacy regulations. PMID- 11478184 TI - Major U.S. disability and compensation systems graphically compared. AB - This article offers the reader a "bullet" overview of the principal systems of disability determination. The reader is referred to other sources for more in depth coverage and a detailed historical background and overview of these systems. PMID- 11478185 TI - Goniometric techniques for range-of-motion assessment. AB - The authors hope that this article promotes greater understanding of the methodology available for measuring and recording joint ROM and fosters movement towards the eventual adoption of a single integrated goniometric system for the evaluation and recording of musculoskeletal impairment and disability. Towards this end, the authors have focused on applications of the Neutral-Zero Measuring Method and SFTR documentation and Recording System, and have highlighted the advantages of this system when compared to more conventional approaches. Examination and recording procedures have been described briefly, and examples of appropriate instruments and their applications have been illustrated. The reader is referred to more extensive discussions elsewhere of the measurement techniques themselves. PMID- 11478186 TI - Applications of functional assessment to musculoskeletal disability evaluation. AB - A review of the history of the disability determination for LBP suggests that the early focus upon disability as a direct "effect" of impairment has given way to an updated approach acknowledging the multitude of factors, many of which are subjective, that contribute to an individual's future return to work and disability status. In addition to the evaluation of the underlying impairment and pathology, physicians performing disability evaluations for LBP will be best served when they can assess the individual's functional status, and incorporate that information into their decision regarding the individual's current limitations and prognosis. As discussed above, many of the factors contributing to the evaluation of functional status, including pain, physical functioning, and affective status, are necessarily subjective. Nonetheless, the rehabilitation literature suggests that inclusion of this information into the disability determination using a patient-centered approach may provide the factors that are most potent with respect to patient prognosis and eventual return to work. PMID- 11478187 TI - Evaluation and reporting requirements of the disability examiner. AB - Critics of the system within which IMEs occur might argue that there is in fact no such entity as a truly impartial evaluation because the practitioner always is aware of the source of payment for the visit and will color opinions in favor of that payor. Nonetheless, the ethical practitioner has no problem whatsoever refuting that rather cynical attitude. First, payment for any type of clinical visit must come from somewhere. Most importantly, always keep in mind what is in the best medical interest of the person you are examining, and you will avoid virtually all ethical dilemmas. To illustrate obvious examples, if a claimant has the objective impairments of herniated disc with associated radiculopathy, it makes no medical sense, and would certainly be unethical, to allow unrestricted RTW to a physical job. That clearly would not be in the best interest of a patient you were treating, nor should that be your opinion for an IME. Similarly, a person without any objective impairment documented after careful search is not well served by a continuing program of medications, time off work, useless modalities, and a growing "sickness" frame of mind. That would definitely not be in their best interest, and your opinion for a treatment case or for an IME should reflect that without hesitation. Once you make a consistent habit of always invoking what you truly believe to be in the best medical interest of the examinee, the long-term interests of the other involved parties will also be well served as a consequence. The insurance company will not be asked to either continue paying treatment and wage replacement costs for someone who could safely work, nor will it be forced to accumulate the unnecessary expenses of recurrent injuries to an injured worker returned to work prematurely. By faithfully adhering to the simple principle of not swaying from the best interest of the examinee, you might lose referral business from some sources. Certain employers, case managers, insurance carriers, attorneys, or others might want to rely on your opinion in all cases, perhaps to conclude that claimants are rarely if ever impaired (or at least not from work or auto accident-related events). Your practice and your reputation are much better off without that business. A well planned and carefully performed IME, resulting in a detailed, understandable, and defensible report with the elements outlined in this article, will serve you and your referring sources well. PMID- 11478188 TI - Evaluating pain and disability. AB - This article has attempted to present an overview of how pain and disability interact, the incidence and cost of disability secondary to painful conditions, a review of the current disability systems, how pain should be evaluated, and some thoughts on the future of disability and pain. Most people are unhappy with the present process from an administrative, logistical, and cost-effective point of view. However, there is a continuing concern that change will be driven largely by cost containment rather than other important aspects of disability. PMID- 11478189 TI - Promoting ethical and objective practice in the medicolegal arena of disability evaluation. AB - As providers of medical information and testimony, clinicians have ultimate responsibility for ethical conduct as it relates to this information. The authors offer the following recommendations for enhancing ethical relationships between expert clinicians and the courts. 1. Avoid or resist attorney efforts at enticement into joining the attorney-client team. Such compromises of scientific boundaries and ethical principles exist on a continuum ranging from standard attorney-client advocacy at the beginning of the expert consultation phase (e.g., promotional information at the forefront of retaining an expert, with either provision of selective or incomplete records or less than enthusiastic efforts to produce all records) and extending to completion of evaluation, when requests for changes in reports and documentation might be made. 2. Respect role boundaries and do not mix conflicting roles. Remember that the treating doctor possesses a bond with the patient but does not as a rule obtain complete preinjury and postinjury information in the context of assessing causality and apportionment. In contrast, the expert witness must conduct a thorough and multifaceted case analysis sans the physician-patient relationship in order to facilitate objectivity and allow optimum diagnostic formulations. Finally, the trial consultant's function in this adversarial process is to assist with critically scrutinizing and attacking positions of experts for the opposing side. These roles all represent inherently different interests, and mixing them can only reduce objectivity. 3. Insist on adequate time for thorough record review, evaluation, and report generation. Also insist on sufficient time and preparation for deposition and court appearances. 4. Work at building a reputation for general objectivity, reliance on multiple data sources, reaching opinions only after reviewing complete information from both sides, and completing the evaluation. 5. Spend a good amount of time actually treating the patient population being examined or being offered testimony about. This treatment should be current and should be of a similar frequency to treating practitioner specialists. Be able to discuss relevant research and scientific methodology issues competently and without notes. 6. Arrive at opinions only after reviewing all of the evidence from both sides of the adversarial fence, employing multiple data sources, completing the evaluation, and interpreting data within the full context of comprehensive historical, behavioral observation, and contextual information. Being otherwise favorable to retaining attorney interests suggests endorsement of "opinion prostitute," "scientific perjurer," or "hired gun" status. The only way a practitioner can reduce the likelihood of facing an "opinion prostitute" on the opposing side in future cases is to insist on establishing and maintaining a reputation for scientific objectivity. 7. Balance cases from plaintiff and defense attorneys. Predilection for one side or the other suggests bias and sets up predisposition to nonobjectivity. For example, a preponderance of plaintiff work suggests an overdiagnosis or uncritical sympathy bias, whereas a ratio that favors hiring by the defense suggests an underdiagnosis or skepticism bias. Perhaps Brodsky's suggested cut-off ratio of .8 for favorability findings would represent an initial cutoff for defense versus plaintiff ratio. That is, experts should do at least 20% work for the opposite side of the current case being represented. Further, it might be a reasonable expectation that data on these ratios be collected as an important method for ensuring objective opinions. 8. Ensure against excessive favorability to the side of the retaining attorney or firm. Objectivity demands that scientific opinions not be influenced by the position of the legal advocate. Importantly, Brodsky recommends using a ratio of .8 as a cut-off for detecting excessive bias. That is, practitioners should possess prerequisite objectivity to disagree with the referring attorney at least 20% of the time. We suggest that a more useful cut off would be .75, where experts are expected to generate findings that do not support the referring attorney's position at least 25% of the time. 9. Never arrive at opinions that are inconsistent with plaintiff records, examination data, test data, behavioral presentation, and so forth, especially when such opinions are favorable to the side of the retaining attorney firm. Instead, use the following recommendations. (ABSTRACT TRUNCATED) PMID- 11478190 TI - Disability evaluation following traumatic brain injury. AB - Accurate disability evaluation of a patient with TBI is a very difficult and detailed process. It requires an excellent background concerning the evaluation of all the physical, cognitive, behavioral, and functional abnormalities associated with TBI. Texts that highlight all these abnormalities include Medical Rehabilitation of Traumatic Brain Injury by Horn and Zasler and Rehabilitation of the Adult and Child with Traumatic Brain Injury by Rosenthal et al. In addition, appropriate disability rating can only be performed by a physician with expert skills in obtaining accurate historical information and completing a detailed physical examination. Often, the historical information must be obtained from many sources because the patient may supply inaccurate information because of his or her cognitive deficits. Interviews with family members, caregivers, therapists, friends, and employers are sometimes necessary to obtain an accurate historical picture. Premorbid functioning, behavior, and personality are important because previous abnormalities are often exaggerated after the TBI. The physical examination should be tailored to provide detailed objective information concerning all deficits identified through the history. If cognitive and behavioral problems are identified through either the history or examination, a neuropsychologic assessment is necessary. All this information should be available before the disability or impairment rating. Only with detailed information can a clinician provide an accurate rating. PMID- 11478191 TI - Determining impairment following spinal cord injury. AB - Determining the level of impairment and disability of an individual with SCI is reasonably straightforward. An impairment rating may be determined by either analyzing the impact of the SCI on various bodily systems or by considering the injury as a more global diagnostic category. Functional abilities based on level of neurologic preservation are well recognized. Although secondary medical complications, which may affect both impairment and disability, can arise at any time after SCI, neurologic and functional abilities are overwhelmingly stabilized by 12 months postinjury. A comprehensive history and physical examination should allow even the inexperienced examiner to obtain a valid determination of impairment after SCI. Although a more functionally oriented and perhaps interdisciplinary evaluation is needed to assess disability, it is also fairly straightforward. On the other hand, an evaluation of handicap is a more challenging undertaking, requiring a more detailed knowledge of the field of vocational rehabilitation. PMID- 11478192 TI - Disability evaluation following stroke. AB - The physician performing a disability evaluation needs to document the stroke patient's impairments carefully. This is achieved through a thorough history, physical examination, and if necessary, neuropsychologic assessment. Most often, the clinician will deal with the Social Security Administration system; however, the physician may occasionally need to provide this information for private disability insurance. PMID- 11478193 TI - Disability evaluation and impairment rating for the person with an amputation. AB - Impairment rating for the person with an amputation requires a total body assessment in addition to the amputated extremity. In particular, those systems essential for prosthetic wear and function, skin coverage, sensation and motor function, and proximal joint range of motion should be included. Spinal function, pain, and psychologic impairment are frequently involved as secondary issues related to the amputation. All of these systems must be assessed and a determination made whether permanent impairment is present. The final calculation of the impairment rating uses the appropriate body systems and worksheets. Most persons with an amputation will have some appropriate work restrictions applied and all will have future medical needs. PMID- 11478194 TI - Disability evaluation following burn injury. AB - This article provides an overview of impairment rating in the burn survivor. It is not intended to be a complete guide; each jurisdiction and each agency requires the physician to use the necessary required criteria. This article also emphasizes the significant rating differences between agencies and jurisdictions and in the latitude given the health professional in performing the rating. PMID- 11478195 TI - Disability evaluation of extremity fractures. AB - Most fractures heal with restoration of bone and joint function. Disability is generally temporary in these instances, and there is no permanent impairment to be rated in the majority of cases. However, multiple complications can occur that can lead to loss of skeletal function, restricted range of motion, and associated soft tissue injuries, and neurovascular damage can dramatically and severely compromise function and performance. Clearly, not all fractures heal within the timeframes as outlined in Table 2, and determination of the point of maximum medical improvement, which must be achieved in order to declare an impairment as permanent, can be challenging in these cases. Some individuals with fractures that demonstrate bone union radiologically may develop long-term disability; nearly 30% of individuals with a unilateral lower extremity fracture will not return to work within 12 months of injury. In the absence of a complicating soft tissue injury, the explanation for this discrepancy between impairment and disability can be elusive. During the past 10 years, improved systems of trauma care have reduced mortality rates in the United States. As greater numbers of persons survive motor vehicle accidents and other major trauma, the possibility exists that increasing numbers of fractures leading to impairment or disability may be seen. Recognition of how and when fractures heal, possible complications that may arise, risk factors for long-term disability, and the correlation of vocational and avocational physical demands with the resultant outcome will enable the clinician to appropriately and accurately assess disability status, design a rehabilitation program, and assign an impairment rating. PMID- 11478196 TI - Impairment rating of the cardiovascular system: Idiopathic cardiomyopathy. AB - Disease of the cardiovascular system is the leading cause of morbidity and mortality among the adult population in the United States. Physicians performing impairment ratings attributable to cardiovascular disease are required to correlate the extent of the disease and its impact on a person's daily activities. A person's final impairment determination may vary based on the specific guidelines used (American Medical Association Guides to the Evaluation for Permanent Impairment, Social Security, State Worker Compensation requirements, etc.), the version of the guidelines (year, edition), and each guide's specific criteria. The examiner needs to know which rating system to use, the system's criteria, critical definitions, and potential limitations. PMID- 11478197 TI - Impairment rating and disability evaluation of the pulmonary system. AB - An increasing number of patients are being seen by physicians for impairment and disability of the pulmonary system, partially because of the increasing prevalence of chronic obstructive pulmonary disease and increased awareness of industrial and environmental hazards to the respiratory system. As legislative efforts and social expectations towards entitlement and compensation become more permissive, an increasing number of individuals with respiratory impairment are claiming disability and seeking compensation. Consequently, the physician whose practice includes pulmonary rehabilitation may be expected to evaluate and rate impairment and disability of the pulmonary system. Fortunately, for respiratory disorders, excellent objective measures of pulmonary function exist that are readily applicable to the process of disability assessment. This article highlights the diagnostic procedures and assessment criteria of choice for pulmonary disability determination. PMID- 11478198 TI - Impairment and disability rating in low back pain. AB - LBP is one of the two most common forms of disability in Western society (mental illness is the other), and remains a thorny problem in the arena of disability evaluation. Disability evaluation after LBP differs whether the pain is work related or not. If work-related, guidelines for disability evaluation differ by jurisdiction and type of employment (e.g., private vs. federal employee). When outside of the workplace, thresholds for disability differ between entitlement programs (Social Security Disability) and private insurance programs (long-term disability insurance). In the patient without obvious findings, the disability evaluating physician needs to be caring and compassionate and yet maintain an objective stance with the understanding that there may be significant psychosocial overlay in patients with nonobjective pain complaints. Although some would argue that objective independent medical evaluation is an oxymoron, psychiatrists have excellent training and perspective with which to do so. The patient suffering from catastrophic brain injury or spinal cord injury offers a useful contrast--on the most severe end of the disability spectrum--to the patient with persisting low back complaints but normal physical examination. As a society, we have to wisely manage the funds that comprise our social "safety net" in order to provide for persons with severe disability who cannot provide for themselves. It would then follow that patients with minor impairments/disabilities should receive minor (i.e., noninflated) ratings. Psychiatrists need to enable rather than disable their patients. PMID- 11478199 TI - Disability issues with carpal tunnel syndrome. A case discussion. AB - This article reviews nerve entrapments and cumulative trauma disorders of the upper limbs including a problematic carpal tunnel syndrome (CTS) patient case, idiopathic arm pain, and ulnar neuropathy. Major points of the discourse focus on the problems of treatment, especially as they relate to impairment and disability management. PMID- 11478201 TI - The physiatrist as preferred disability specialist. PMID- 11478200 TI - Disability evaluation of fibromyalgia. AB - These cases represent individuals who feel they have a severe impairment and are "disabled." They have been labeled with fibromyalgia. They are truly distressed. Their symptoms, their courses, are more chronic and refractory than those of medically ill patients, and they are high users of medical services, laboratory investigations, and surgical procedures. These patients see multiple providers simultaneously and frequently switch physicians. They are difficult to care for, and they reject psychosocial factors as an influence on their symptoms. Such persons "see themselves as victims worthy of a star appearance on the Oprah Winfrey show. A sense of bitterness emerges...." Shorter, a historian, believes that fibromyalgia is "heaven-sent to doctors as a diagnostic label for pain patients who display an important neurotic component in their illness. Our culture increasingly encourages patients to conceive vague and nonspecific symptoms as evidence of real disease and to seek specialist help for them; and the rising ascendancy of the media and the breakdown of the family encourage patients to acquire the fixed belief that they have a given illness...." Regarding the finding of "disability," this is a social construct, and many authors believe it is society and the judicial system who must decide who can work. To remain objective, the physician should report the objective clinical information. Physicians need not and should not sit in judgment of the veracity of another human being. PMID- 11478202 TI - Sexual activity and contraceptive practices among teenagers in the United States, 1988 and 1995. AB - OBJECTIVES: This report presents national estimates of sexual experience, contraceptive use, and selected aspects of sexual behavior among never-married males and females aged 15-19 years in the United States. Data are presented for the years 1988 and 1995 according to age, race and Hispanic origin, progress in school, and other relevant characteristics. Tables present trends over time as well as comparisons between subgroups. METHODS: Descriptive tables of numbers and percents are presented and interpreted. Data for females are from the National Survey of Family Growth, and data for males are from the National Survey of Adolescent Males. RESULTS: About half of all never-married teenagers, about 17.5 million, had had sexual intercourse at least once in 1995. For male teenagers, this represents a decline since 1988, and for females, the proportion was stable across the two time points. The proportion of teen females who had sex before age 15 years increased. In 1995, 29 percent of females and 19 percent of males had unprotected recent sexual intercourse. About one-quarter of teens used no contraceptives during their first sexual intercourse. The condom remained the most popular method of contraception. Although teenagers' use of oral contraceptives dropped between 1988 and 1995, use of injectable and implant contraceptives began. Teenagers with more highly educated mothers, mothers who delayed their first birth beyond age 19 years, those from two-parent families, and those whose schooling was on schedule, were less likely to engage in sexual risk behaviors. These teenagers, along with those who were Protestant, also experienced the largest improvements across time in sexual risk behaviors. PMID- 11478203 TI - [Don't trust blindly computerized tests! "A not completely perfect test by a competent professional is better than a perfect test by an amateur"]. PMID- 11478204 TI - [A report from the United States: women with myocardial infarction don't receive the same attention as men with myocardial infarction]. PMID- 11478205 TI - [Using projective tests in forensic psychiatry may lead to wrong conclusions. Only empirically tested tests should be used]. AB - The use of projective and psychometric psychological tests at the Department of Forensic Psychiatry in Stockholm (Huddinge), Sweden, was studied for a population of 60 men, including many patients with neuropsychological disabilities and multiple psychiatric disorders. The results showed that the use of projective tests like Rorschach, Object Relations Test, and House-Tree-Person was more frequent than the use of objective psychometric tests. Neuropsychological test batteries like the Halstead-Reitan Neuropsychological Test Battery or Luria Nebraska Neuropsychological Battery were not used. The majority of patients were, however, assessed by intelligence scales like the WAIS-R. The questionable reliability and validity of the projective tests, and the risk of subjective interpretations, raise a problem when used in a forensic setting, since the courts' decisions about a sentence to prison or psychiatric care is based on the forensic psychiatric assessment. The use of objective psychometric neuropsychological tests and personality tests is recommended. PMID- 11478206 TI - [Childhood amnesia and emotional trauma. Easiest to prompt the smallest children to provide erroneous details concerning abuse]. AB - A basic understanding of children's memories for emotional trauma is of importance for physicians and other health care professionals. The aim of this paper is to describe recent research and discuss important dilemmas within this field. The ability of infants and toddlers to verbally describe autobiographical memories is for several reasons limited. Very early experiences of e.g. intense pain can leave traces in the nervous system but of a kind which is not specific enough to allow retrospective conclusions about the nature of these experiences. Research during the past decades has relatively consistently demonstrated that pre-school children are more vulnerable to suggestion including abuse-related suggestions than are older children and adults. However, many children who have been subject to sexual abuse seem to maintain a lifelong silence about their experiences. The use of autobiographical testimony from young children is more complex and risky than many professionals believed about a decade ago. Establishing the knowledge derived from recent years' research in this area as a basis for clinical practice is one of the most important tasks for the future. PMID- 11478207 TI - [A vicious circle when headache medication is the cause of chronic headache. A well-planned ambulatory detoxication can result in dramatic improvement]. AB - Chronic daily headache is a commonly encountered syndrome. Typically affected are persons who as a result of recurrent attacks of migraine or tension-type headache have used medications regularly during an earlier stage. Based on clinical experience it is concluded that outpatient drug-withdrawal should be a first-line treatment for patients with chronic daily headache and frequent long-term drug use. If there is no improvement treatment with a tricyclic drug is recommended. PMID- 11478208 TI - [Joggers live longer]. AB - In all 4,658 men, 20-79 years of age, with no prior myocardial infarction, attended two examinations. Jogging status was self-reported based on the question "Are you a jogger?" The study shows that regular jogging is not associated with increased mortality in men, as the joggers had significantly lower mortality than non-joggers. The lower mortality of joggers could be an effect of the physical training, but it could also be due to selection or a combination of both. On the other hand, numerous studies within the fields of epidemiology, work-physiology, psychology and biochemistry have all pointed toward a beneficial effect of physical activity on health, but the optimal intensity, frequency and duration of physical activity has yet to be established. PMID- 11478209 TI - [Immunological changes in hypertension and atherosclerosis]. PMID- 11478210 TI - [X-ray examination of the spine can intensify patient's experience of disease]. PMID- 11478212 TI - [Leadership, communication and respect in health care]. PMID- 11478211 TI - [A new test for screening of neuropsychological disorders]. PMID- 11478213 TI - [Self-interest can be of benefit for everyone and save medical research]. PMID- 11478214 TI - [Development of better software requires close cooperation between physicians and technicians. Some thoughts on current rudimentary computer technology in primary health care]. PMID- 11478215 TI - [Occupational stress, suicide and fatigue depression]. PMID- 11478216 TI - [Ambulance personnel should take pictures at the site of accidents!]. PMID- 11478217 TI - [One more time--evidence-based is not the same as randomized trials!]. PMID- 11478218 TI - [Is Basaglia responsible for a better psychiatry? On the biography of the Italian psychiatric reform]. PMID- 11478219 TI - [Somatoform disorders. Historical development and modern diagnostic conceptualization]. AB - Patients with "medically unexplained" bodily symptoms form a major group in the medical care system. Diagnostic and therapeutic problems very often arise, and the concept of "somatization" promotes a better understanding of this patient group. Important historical stages in the development toward a modern diagnostic classification of somatoform disorders are outlined. A critical comment on the diagnostic approaches according to DSM-IV and ICD-10 follows. PMID- 11478220 TI - [Status and development of psychiatric care in Italy. 22 years after Franco Basaglia's reform regulation no. 180]. AB - Since reforms were introduced in 1978, the treatment of mental illness in Italy is less uniform than ever before. The essential and core aspects of these reforms were not implemented in many southern provinces, but there are flourishing private clinics in Rome and other metropolitan areas, and in the highly developed regions of northern and central Italy there are institutions meeting very high standards of complementary care. However, reforms in these areas had begun as early as in 1968 in accordance with Act 431 passed that year, and in these regions the newer reforms functioned only as a catalyst. The extremely high goals set by the "Italian experiment" were not realized, but the achievements of several Italian provinces in the areas of ambulant and supporting psychiatric care can certainly be viewed as exemplary. A general trend toward increased critical examination of ways of dealing with the mentally ill from the medicinal, humanitarian, and social points of view is emerging. PMID- 11478221 TI - [The current status of psychiatric care in Italy]. AB - Psychiatric reforms in Italy have been the subject of many publications since 1978, when these reforms became law. One such publication is the article by Hinterhuber et al. entitled "Status and Development of Psychiatric Care in Italy" appearing in this edition of Der Nervenarzt [17]. In it, a number of unsolved issues are pointed out, concentrating on some of the main problems encountered by any country going through a process of major institutional change. Some additional points are helpful to a discussion of Italian psychiatric methods and their implications for reorganizing psychiatric practice elsewhere in Europe. Making use of certain "key concepts," we review these questions [2]. PMID- 11478222 TI - [Single sex or mixed sex treatment in the psychiatric clinic from the viewpoint of patients]. AB - Patients in a psychiatric hospital were surveyed by questionnaire to evaluate their preferences on separate or mixed-sex treatment. The adjusted response rate was 74.5%. About 75% of the responders preferred a mixed ward and about 25% preferred a single-sex ward. Females more frequently chose the type of ward to which they had been allocated. By contrast, males were in favour of mixed wards, independently of their present type of ward. Younger patients, voluntarily admitted patients, and those with substance addiction preferred mixed-sex wards. After mixing the sexes in formerly single-sex wards, a significant increase in preference of mixed-sex wards was found, due to increased affirmation by female patients. However, about 9% of women and men preferred single-sex treatment. Two open general psychiatric wards, each with 20 beds and separating females and males, could be established in psychiatric hospitals to satisfy this preference. PMID- 11478223 TI - [Depressive disorders and antidepressive therapy. A comparison of neurology practice and psychiatric clinic]. AB - It is a question of great importance when psychiatric patients should be treated as inpatients or as outpatients. This decision depends on therapeutic as well as economic considerations. However in depression, as in other mental illnesses, many cases could reasonably be treated as in- or outpatients alike. Despite the importance of this question, there is to our knowledge no epidemiological study which compares patients in inpatient and outpatient settings within the current German medical system. 1300 inpatients and 5376 outpatients were investigated using identical methodologies within a drug utilization study of the antidepressant venlafaxine. Inpatients were found to be more severely ill and have a recurrent course of illness and higher rates of comorbidity. Outpatients more often had a chronic course. Inpatients were treated more often and with a wider variety of medication than outpatients, with the exception of phytopharmaceuticals. Venlafaxine was given in higher doses in the inpatient setting and combined more often with other psychotropic medications. Complaints of adverse drug reactions were comparable in both groups. Nausea was the most frequent, reported by 9.7% of inpatients and 15.2% of outpatients. The data show that the distribution of patients in outpatient and inpatient settings corresponds to the stepwise structure of the German health care system. PMID- 11478224 TI - [Socio-familial and epilepsy specific stress on women with epilepsy]. AB - The Berlin longitudinal study on teratogenic effects of maternal epilepsy was conducted on children in 1976. Approximately 18 years later, 103 of these children's mothers were reassessed with regard to somatic, psychological, and social well-being or impairment. The families with previously diagnosed epileptic mothers (n = 59) and control families (n = 44) did not statistically differ in rates of family separation, divorce, or unemployment of the partners. There were also no significant differences in the mothers' current psychological quality-of life status (SF-36) or their beliefs on internal or external disease control (FKK). There were, however, group differences in more closely illness-related aspects. In spite of an appreciable remission rate of 28% over the past 17 years, significantly more mothers in the epilepsy group still reported physical infirmities (GBB), reduced body-related quality of life (SF-36), and more family stress events (FAI). Their self-esteem and competence-related beliefs (FSKN) were significantly lower. They also tended to have a lower level of education or professional training, and some had never been employed. When measuring quality of life, severity of seizure (NHS3) and early epilepsy onset are the major risk factors, regardless of whether one uses epilepsy-specific (QOLIE-10) or general quality-of-life questionnaires (SF-36). PMID- 11478225 TI - [Speech development and intelligence in autism. How uniform is Asperger syndrome?]. AB - Since the introduction of a separate diagnosis for Asperger's syndrome in the ICD 10 and DSM-IV classification systems, a controversial debate has continued on whether Asperger's syndrome is a specific, clearly distinguishable disorder within the autistic spectrum or whether it represents a milder phenotypical variation of autism. The effect on the amount of autistic symptoms of the variables language delay and level of intelligence was analyzed within a sample of individuals exhibiting autism diagnosed by standardized methods. Both variables showed a significant effect on the degree of autistic symptoms in that impairments in social interaction were less noticeable. In addition, a subsample of individuals exhibited symptoms assumed to be characteristic for Asperger's syndrome. The findings support the assumption that autism and Asperger's syndrome represent "extreme points" on a scale of severity, which leads to the suggestion that the classification of different subtypes of autism could be abandoned in favor of a dimensional (multiaxial) approach. PMID- 11478226 TI - [A patient with Gulf war syndrome. On discussion of an uncertain disease picture]. AB - The Gulf War syndrome represents neurological and neuropsychological disorders in veterans of the Persian Gulf war. Until today, the various symptoms observed could not be attributed to any defined disease. As a possible cause, exposure to neurotoxic agents such as the organophosphates used during the war has been suggested by many authors. We report on a 29-year-old man who suffered from dysmnesia, disturbance of orientation, cognitive impairment, and double vision. His history revealed several front-line operations in 1990 and 1991 during the Gulf War. Physical examinations showed a complex eye-movement disturbance and a horizontal nystagmus, which was neuro-ophthalmologically confirmed. The early auditory potentials referred to a brainstem dysfunction and the cognitive disturbances correlated to changes in the late-appearing component of event related potentials (P 300). Brain imaging with CCT, MRI, SPECT, PET, and EEG and CSF showed no pathologies. Neuropsychological tests disclosed severe cognitive impairment especially concerning memory. Three-month follow-up studies in a department of psychosomatic medicine excluded a dissociative disorder as a feature of a post-traumatic stress or a conversion disorder. This is the first case of Gulf War syndrome in Germany. We focus on an unfamiliar complication after the war. The recent literature is reviewed. PMID- 11478227 TI - [Hoigne syndrome. Case report and current literature review]. AB - Hoigne's syndrome is currently considered a pseudoanaphylactic or pseudoallergic reaction following intramuscular and aqueous procaine penicillin administration. This disorder is characterized predominantly by neuropsychiatric alterations including severe psychomotor agitation with confusion, sensations of disintegration, depersonalization, and derealization, perceived changes of body shape, visual and auditory hallucinations, panic-like anxiety including fear of death as well as alterations of consciousness and seizures. Beside the "classic" immediate manifestation of Hoigne's syndrome, subacute forms as well as reactions of the so-called latent type are also known. Including a typical case report, we present a review of the currently available literature concerning clinical picture, hypotheses on origin, and possible therapy regimens of this underdiagnosed complication of antibiotic penicillin treatment. PMID- 11478228 TI - [Neurobiological findings in a patient with 48-hour rapid cycling bipolar affective disorder. A case report]. AB - Forty-eight-hour rapid cycling is a rare subclass of affective disorders and entails a regular periodic change of moods. The following case study describes the symptoms, therapy, and outcome of a patient suffering from daily switching between mania and depression along with neurobiological findings correlated to the affective cycles. We found alternating reduction and prolongation of sleep duration during manic and depressive days as well as differences in the amount of REM sleep. Cortisol secretion was regularly increased during depressive days. Regarding thyroid-stimulating hormone (TSH) secretion, the circadian rhythm was absent on depressive days. However, the glucose metabolic rate as measured by positron emission tomography (PET) did not differ on manic and depressive days. The patient reached almost complete remission under treatment with lithium. PMID- 11478229 TI - [Hypernatremia as a rare complication in change from lithium to valproate]. AB - This paper is about a female patient who suffered from bipolar disorders for several decades. Due to a change in medication from lithium to valproic acid by infusion, the patient fell into a coma. Hypernatremia, which had developed unnoticed, was later discovered as the reason. This hypernatremia was due to renal diabetes insipidus stemming from the decades of lithium therapy, with accompanying polydipsia and polyuria, and also to the failure to give enough fluid to the somnolent patient. PMID- 11478230 TI - [Bulimia nervosa in homosexuality and HIV infection in the man. Case report]. AB - A male with bulimia nervosa is reported whose homosexual behavior resulted in HIV infection. The goal of this case study is to shed light on the course characteristics of bulimia nervosa occurring together with HIV infection in connection with homosexuality. The patient had experienced a number of traumas including sexual abuse, which surely had an influence on his developing bulimia nervosa. His longing for slimness, shown by the excessive preoccupation and dissatisfaction with body shape and weight may be considered typical for bulimia nervosa in homosexual men. Furthermore, male subjects with eating disorders often seem to be overweight prior to the start of bulimia nervosa, which also was the case in this subject. The course of his HIV infection so far has had no effect on severity of the bulimic symptoms. However, his bulimia nervosa apparently did had negative effects on the course of the HIV infection. PMID- 11478231 TI - [Loperamide abuse in anxiety disorder]. AB - Anxiety disorders often accompany somatic correlates of anxiety such as abdominal trouble and diarrhoea. Nevertheless, misuse of antidiarrhoeals has been described only once. We report on a 34-year-old woman who abused loperamide for over 10 years, taking up to 16 mg per day. Loperamide is an opioid that stimulates opioid receptors in the gastrointestinal tract, thereby inhibiting intestinal secretion and propulsive peristalsis, probably without affecting the central nervous system. As our patient's digestive trouble was a consequence of her states of anxiety, diazepam application helped against her diarrhoea as well. This second strategy of self-medication gained particular importance for her, when even high doses of loperamide stopped having an effect. So she developed a benzodiazepine dependence. As soon as she began treatment as an inpatient, her feelings of anxiety became less and the diarrhoea disappeared. PMID- 11478232 TI - [The "qualified outpatient detoxification" model project]. AB - In 1998, a model project of outpatient detoxification in alcoholics was initiated in a German outpatient clinic. Inclusion and exclusion criteria of this project, which is supported by local health care providers, are presented. Between August 1998 and June 2000, 205 patients were screened. Psychotherapeutically motivating elements are integrated, so that this may be termed a "qualified" detoxification program. One hundred forty-one patients entered the program, of whom 127 (90%) successfully finished treatment. No serious medical complications were observed. Of those who completed treatment, 121 entered further alcoholic rehabilitation programs. Some preliminary data of an ongoing catamnestic study suggest that, after a mean of 10 months post-detoxification, 61 (50%) of these patients are still abstinent and in rehabilitation programs. The findings suggest that outpatient alcohol detoxification is safe and efficient, at least for a subgroup of patients, and also indicate the possible efficacy of concomitant psychotherapeutic interventions during this mode of treatment. PMID- 11478233 TI - [Mental competence for informed consent as a relational model--clinical practice and medical ethics analysis. Comment on the contribution by J. Vollman]. PMID- 11478234 TI - [Comment on continuing education by L. Pezawas, M. Stamenkovic and S. Kasper: Recurrent brief depressive episodes--epidemiology, clinical aspects, diagnosis and therapy]. PMID- 11478235 TI - Cost effectiveness of reducing dioxin emissions from municipal solid waste incinerators in Japan. AB - The main sources of dioxin emissions are municipal solid waste incinerators. The Japanese national government has set an emission standard for dioxins to reduce dioxin exposure levels. In this study, cost-effectiveness analyses are carried out regarding countermeasures that were recently taken and are being taken at municipal solid waste incinerators in Japan. Annual costs were estimated by telephone survey and model calculations. Annual decrease in the incidence of cancer was estimated in three steps. First, the annual decrease in the volume of dioxin emissions was estimated. Next, using a mathematical model, the annual decrease in human exposure was estimated. Finally, the annual decrease in the incidence of cancer was estimated by applying the cancer slope factor. When annual costs are divided by the annual number of life-years saved, cost per life year saved (CPLYS) was obtained. CPLYS was estimated to be 7.9 million yen for emergency countermeasures and 150 million yen for long-term countermeasures. However, it must be noted that these obtained CPLYSs are highly dependent on the cancer slope factor and should be considered as an upper limit since there may be a cancer effect threshold. PMID- 11478236 TI - Proposal for including what is valuable to ecosystems in environmental assessments. AB - Assessment scientists and managers depend on social values to identify the goals that will be used to guide environmental assessments. These goals are commonly identified by examining the vested interests of the various social groups that are stakeholders in a region. However, knowledge about what people value represents only part of the information needed to identify comprehensive assessment goals for environmental systems that include both economic and ecological components and processes. All parties also need to understand what is valuable to ecosystems because that determines the ecological patterns and processes that prevail in the long run. The competition among alternate system designs for available energy determines the viability of the choices that people make for their environment. Ecosystems that prevail in competition use the process of self-organization to create system designs that maximize the use of ever-changing sources of available energy. The efficacy of ecosystem designs can be evaluated using the maximum empower principle, which states that ecosystems evolve toward designs that maximize empower (energy use per unit time). Energy is an accounting quantity that normalizes the different kinds of energy developed in a system so that they may be compared. The counter-intuitive and sometimes controversial results that come from energy analyses are illustrated by examining three environmental problems on the interface between ecology and economics. A process for identifying and using social and ecosystem values to guide environmental assessments is proposed using a conceptual energy systems model that shows how these processes might interact within a region. The probability of realizing a given change in system empower production is suggested as a decision criterion that can be used by managers to evaluate the efficacy of alternatives. PMID- 11478237 TI - Snowmobile contributions to mobile source emissions in Yellowstone National Park. AB - Increases in the number of winter visitors to Yellowstone National Park during the past decade have raised concerns over the environmental impacts of snowmobiling in the park. During the 1998-99 season, more than 62,000 snowmobile and 1300 snow coach trips entered the park. Using the University of Denver's vehicle exhaust remote-sensing equipment, 1385 measurements of carbon monoxide (CO) and hydrocarbon (HC) emissions were collected from in-use snowmobiles at the west and south entrances to the park. Overall means of 392 +/- 4 g CO and 237 +/- 1 g HC were observed per kilogram of fuel consumed. In addition, using an ultraviolet monochromator, 460 measurements of toluene emissions were collected with a mean of 39 +/- 1 g toluene/kg of fuel. Using these data, a mobile source emissions inventory based on fuel use for Yellowstone National Park shows that snowmobiles account for 27% of the annual emissions of carbon monoxide and 77% of annual emissions of hydrocarbons using an equivalent best estimate for the summer mobile source emissions. Use of oxygenated fuels in snowmobiles was found to reduce CO emissions by 13.2 +/- 6.5% without an observed effect on HC emissions. Liquid-cooled sleds were found to have HC emissions 9.5 +/- 2.2% higher than those from fan-cooled sleds because of the increased intake and exhaust port sizes required in the larger liquid-cooled engines, which increases blowby in the 2-stroke engines. PMID- 11478238 TI - PCDD/F deposition time trend to Esthwaite Water, U.K., and its relevance to sources. AB - PCDD/Fs were measured in 16 sections of a sediment core from a freshwater lake in rural England. Each section represented 10 yr of deposition. Concentrations greater than the limit of detection were observed for most homologues in all samples. Three eras of PCDD/F input were identified by their distinct homologue and isomer profiles. sigma(4-8)CDD/F levels in core sections deposited before 1900 were in the order of 100 pg/g dry weight (dw) as compared to 1500 pg/g dw in the most recently deposited sediment. Local industries such as mining, quarrying, charcoal burning, and iron smelting appear to have had a minor impact on the PCDD/F deposition in the lake. Since 1900, two major peaks in PCDD/F input to the lake were evident. The first, reaching a maximum in the 1930s, had an unusual homologue pattern dominated by high molecular weight PCDFs, and the source of this input is unknown. The second, with a maximum in the 1970s, is in keeping with previously reported time trends for Europe and North America. Pre-1900, the TCDD/F isomer pattern was dominated by dimerization products of 2,4 dichlorophenol. Concentrations of P(1-3)CDFs appeared to be connected with the input that peaked in the 1930s, while highest concentrations of P(1-3)CDDs were found in the deepest sediment sections. Concentrations of diCDDs were observed to have increased again over the most recent decades to levels similar to pre-1900 sections. Despite detailed knowledge of the catchment and of industry in the surrounding area, the identity of some sources and the contribution of other known sources remain unclear for each era. This indicates that there remain significant gaps in our understanding of PCDD/F sources and undermines our ability to predict future trends in PCDD/F emissions. PMID- 11478239 TI - Identification of UV photoproducts and hydrolysis products of butachlor by mass spectrometry. AB - The photoproducts and hydrolysis products of butachlor in water were identified by gas chromatography/mass spectrometry. When exposed to UV light, butachlor in aqueous solution was rapidly degraded, giving at least 11 photoproducts as a result of dechlorination with subsequent hydroxylation or cyclization processes. The chemical structures of nine degradation compounds were identified on the basis of mass spectrum interpretation and literature data. Major photoproducts are identified as 8-ethyl-1-butoxymethyl-4-methyl-2-oxo-1,2,3,4-tetrahydro quinoline, 2-hydroxy-2',6'-diethyl-N-(butoxymethyl) acetanilide, and a compound related to butachlor. Minor photoproducts are identified as 2,6-diethylaniline; 1 acetyl-7-ethylindole; N-(2,6-diethylphenyl)-N-(butoxymethyl)acetamide; 2-oxo-N (2,6-diethyl-phenyl)-N-(butoxymethyl)acetamide; 1-hydroxyacetyl-2-butoxyl-3 methyl-7-ethylindole; 1-acetyl-2-butoxyl-3-methyl-7-ethylindole; and two compounds with the chemical structure unknown. The half-lives of butachlor UV photolysis were 7.54, 10.56, and 12.22 min in deionized water, river water, and paddy water, respectively. The half-lives of butachlor hydrolysis at pH 4, 7, and 10 were 630, 1155, and 1155 days at 25 +/- 1 degrees C, respectively. A hydrolysis product at pH 4 was identified by GC/MS to be 2-hydroxy-2',6'-diethyl N-(butoxymethyl) acetanilide. PMID- 11478240 TI - PAHs in dated sediments of Ashtabula River, Ohio, USA. AB - Polycyclic aromatic hydrocarbons (PAHs) in dated sediments from Ashtabula River, Ohio, were determined, and a chemical mass balance (CMB) model was used to apportion sources. Three cores (AR-1,AR-2,AR-3) were dated by correlating uranium supported 210Pb peaks with 1964, 1972, 1977, and 1979 maxima in the discharge record for Ashtabula River. These cores had sedimentation rates between 7.1 and 4.4 cm/year, while a fourth (AR-4) exhibited a much higher rate of 27.8 +/- 18 cm/year. The highest PAH concentration was 11,500 ng/g found in layer 6 of AR-1 (1986), and the lowest was 621 ng/g found in layer 8 of AR-2 (1982). The source contributions to the total PAH concentrations estimated by the CMB model are 0.1 2.2%, 16.8-22.8%, and 78.1-83.8% for wood burning (WB), coke oven (CO), and highway dust (HWY), respectively. Petroleum generated PAHs have maximal contribution during 1977-79, and wood burning PAHs show minimal emissions during 1975-77 in accordance with U.S. consumption records and other studies. Among six PAH markers, only phenanthrene may be subjected to aerobic biodegradation or photolysis with an apparent half-life of 0.005-0.025 year. No anaerobic degradation was observed based on the CMB model. The model works well for the nonmarker compounds, fluoranthene, and benzo[b]fluoranthene/benzo[k]fluoranthene. PMID- 11478241 TI - Polychlorinated biphenyls in nonaccumulating, century-old sediments: sources, signatures, and mechanism of introduction. AB - This study documents the occurrence of highly chlorinated PCB congeners in stream sediment deposited over 100 years ago. Penta- to heptachlorinated congeners (> 80%) have been found at concentrations up to 78.8 ng/g (dw) in core samples of a small, rural tributary of Lake Ontario. Lower chlorinated congeners and other organochlorine compounds occur sporadically; 210Pb and 137Cs are lacking. The most plausible mechanism is accumulation of dissolved-phase PCBs in permeable sediments adjacent to the creek channel. The similarity between core and air samples collected in the drainage basin suggests derivation from a residual fraction of atmospherically derived PCB congeners. PMID- 11478242 TI - Nonylphenol affects gonadotropin levels in the pituitary gland and plasma of female rainbow trout. AB - Female rainbow trout (Oncorhynchus mykiss) were exposed to 4-nonylphenol (NP) at (mean measured) concentrations of 0.7, 8.3, and 85.6 micrograms/L for 18 weeks, during early ovarian development. Fish were sampled sublethally every six weeks, and terminal samples were taken at 18 weeks. NP induced an estrogenic effect (the synthesis of vitellogenin) at concentrations of 8.3 and 85.6 micrograms/L. An effect on gonadotropin synthesis and secretion was also observed. Plasma follicle stimulating hormone (FSH) levels and FSH gene expression in the pituitary were the most sensitive endpoints assessed, being reduced at the lowest dose employed (0.7 microgram NP/L). Pituitary gland luteinizing hormone (LH) content was significantly lower in fish exposed to 85.6 micrograms NP/L, and LH gene expression was suppressed in fish exposed to 8.3 and 85.6 micrograms NP/L. In contrast, plasma LH concentration increased in these fish, but by a very minor absolute amount, and returned to control levels by the final sampling time. Gonadal development ceased in the fish exposed to 85.6 micrograms NP/L, and steroidogenesis in these fish was also markedly inhibited. Although the mechanisms underlying these responses are unknown, this study demonstrates that NP has adverse effects on pituitary function that can result in inhibition of ovarian development. PMID- 11478243 TI - Reproductive effects of long-term exposure to Bisphenol A in the fathead minnow (Pimephales promelas). AB - Bisphenol A (BPA), a high-volume chemical used to make polycarbonate plastic, epoxy resins, and other chemicals has been reported to be weakly estrogenic. To investigate the effects of long-term exposure to Bisphenol A, a multigeneration study was conducted in which fathead minnows (Pimephales promelas) were exposed to water concentrations of BPA in the range from 1 to 1280 micrograms/L. In this paper, we report the growth and reproductive effects of BPA on sexually mature adults in the F0 generation (after 43, 71, and 164 d of exposure) and the effects on hatchability in the F1 generation. Mean measured concentrations of BPA in the water for all doses, over a 164-d exposure period, were between 70% and 96% of nominal. An inhibitory effect of BPA on somatic growth (length and weight) occurred in adult male fish exposed to 640 and 1280 micrograms/L (after 71 and 164 d). BPA induced vitellogenin synthesis (VTG; a biomarker for estrogen exposure) in males at concentrations of 640 and 1280 micrograms/L after 43 d and 160 micrograms/L after 71 d. In females, plasma VTG concentrations were elevated above controls only after 164-d exposure to 640 micrograms/L. Inhibition of gonadal growth (as measured by the gonadosomatic index) occurred in both males and females at concentrations of 640 and 1280 micrograms/L after 164 d. In males, a concentration of 16 micrograms/L altered the proportion of sex cell types in the testis, suggesting inhibition of spermatogenesis. Concentrations of BPA that induced VTG synthesis and affected gonadal development were lower than those that resulted in discernible effects on reproductive output. Egg production was inhibited at a BPA concentration of 1280 micrograms/L, and hatchability in the F1 generation was reduced at a BPA concentration of 640 micrograms/L (there were not enough eggs spawned in the 1280 micrograms/L group for hatchability studies to be conducted). The results demonstrate that BPA acts as a weak estrogen to fish when administered via the water, with effects on breeding at and above 640 micrograms/L. PMID- 11478244 TI - On-line analysis of reactive VOCs from urban lawn mowing. AB - We measured the release of volatile organic compounds (VOCs) resulting from lawn mowing during continuous ambient air measurements in July and August 1998 in the outskirts of Innsbruck, Austria. These measurements were made with a proton transfer-reaction mass spectrometry system, which allowed simultaneous, on-line monitoring of VOCs in the pptv range. We observed the emission of C6 wound compounds, including (Z)-3-hexenal, (E)-2-hexenal, hexenol plus hexanal, and acetaldehyde immediately following lawn mowing, and a rise in background levels of C6 wound compounds that lasted for several hours. Peak levels of biogenic VOCs following mowing were in the same concentration range (20-60 ppbv) as those originating from combustion engines of lawn mowers, and integrated biogenic emissions were much greater in the drying grass clippings. Additional emissions of acetone and other VOCs resulted from rainfall on these clippings. Since the estimated atmospheric chemical reactivity of VOCs resulting from lawn mowing is of the same order of magnitude as unburned hydrocarbons released during the mowing by gasoline-powered lawn mowers, these biogenic VOCs should be considered in urban air-quality control strategies. PMID- 11478245 TI - Recovery of fish communities in the Finniss River, northern Australia, following remediation of the Rum Jungle uranium/copper mine site. AB - The Finniss River in the wet-dry tropics of northern Australia has received acid rock drainage (ARD) contaminants from the Rum Jungle uranium/copper mine site over more than four decades. Annual-cycle loads of Cu, Zn, Mn, and sulfate, calculated from daily water and flow measurements, have been determined both prior to and following mine-site remediation, that began in the early 1980s. The effects of varying contaminant loads on the relative abundances of seven fish species, sampled by enmeshing nets during dry seasons, were determined by nonmetric multidimensional scaling (nMDS), in combination with cluster-analysis and other nonparametric statistical techniques. These analyses showed that (i) prior to remediation, the impacted region of the Finniss River in 1974 had significantly dissimilar (P < 0.001) and more heterogeneous fish communities, generally characterized by reduced diversity and abundance, compared to sites unexposed to elevated contaminant water concentrations and (ii) postremediation, recovery in fish communities from the impacted region was indicated because they were not significantly dissimilar from those sampled at contemporary (P = 0.16) unimpacted sites, that were also similar to preremedial unimpacted sites. Even though considerable contaminant loads are still being delivered to the impacted region of the Finniss River over the annual cycle, the recovery in fish diversity and abundances is consistent with (a) reductions of in situ contaminant water concentrations at the time of fish sampling, (b) reductions in annual-cycle contaminant loads of sulfate, Cu, Zn, and Mn by factors of 3-7, (c) greatly reduced frequencies of occurrence and magnitude of elevated contaminant water concentrations over the annual cycle, that was most pronounced for Cu, and (d) the absence of extensive fish-kills during the first-flushes of contaminants into the Finniss river proper at the beginning of the wet season, that were observed prior to remediation. As such, the results indicate that there has been ecological benefit to the Finniss River attributable to remedial works undertaken at the Rum Jungle mine site. Recovery in abundances of these fishes may also be due to their time-dependent evolution of tolerance to mine-waste contaminants over their long period of exposure. PMID- 11478246 TI - Siderophore mediated plutonium accumulation by Microbacterium flavescens (JG-9). AB - Uptake of plutonium and uranium mediated by the siderophore desferrioxamine-B (DFOB) has been studied for the common soil aerobe Microbacterium flavescens(JG 9). M. flavescens does not bind or take up nitrilotriacetic acid (NTA) complexes of U(VI), Fe(III), or Pu(IV) or U(VI)-DFOB but does take up Fe(III)-DFOB and Pu(IV)-DFOB. Pu(IV)-DFOB and Fe(III)-DFOB accumulations are similar: only living and metabolically active bacteria take up these metal-siderophore complexes. The Fe(III)-DFOB and Pu(IV)-DFOB complexes mutually inhibit uptake of the other, indicating that they compete for shared binding sites or uptake proteins. However, Pu uptake is much slower than Fe uptake, and cumulative Pu uptake is less than Fe, 1.0 nmol of Fe vs 0.25 nmol of Pu per mg of dry weight bacteria. The Pu(IV)-DFOB interactions with M. flavescens suggest that Pu-siderophore complexes could generally be recognized by Fe-siderophore uptake systems of many bacteria, fungi, or plants, thereby affecting Pu environmental mobility and distribution. The results also suggest that the siderophore complexes of tetravalent metals can be recognized by Fe-siderophore uptake proteins. PMID- 11478247 TI - UV photolysis of nitrate: effects of natural organic matter and dissolved inorganic carbon and implications for UV water disinfection. AB - Nitrite (NO2-) formation during ultraviolet (UV) photolysis of nitrate was studied as a function of pH and natural organic matter (NOM) concentration to determine water-quality effects on quantum yields and overall formation potential during UV disinfection of drinking water with polychromatic, medium-pressure (MP) Hg lamps. Quantum yields measured at 228 nm are approximately 2 times higher than at 254 nm under all conditions studied. In the absence of NOM, NO2- quantum yields decrease with time. With addition of NOM, initial quantum yields increase, and the time-dependent decrease is eliminated. At 15 ppm dissolved organic carbon (DOC) as NOM, the quantum yield increases with time. Dissolved inorganic carbon significantly decreases NO2- yields at pH 8 but not pH 6, presumably by reaction of CO2(aq) with peroxynitrite, a major intermediate in NO2- formation. The results indicate important and previously unrecognized roles for NOM and CO2(aq) in nitrate photolysis. When photolysis was carried out using the full spectrum MPUV lamp and germicidally relevant UV doses, NO2- concentrations remained well below the U.S. maximum contaminant level of 1 ppm N, even with nitrate initially present at 10 ppm N. Under current U.S. regulations, NO2- formation should not pose a significant problem for water utilities during UV disinfection of drinking water with MP Hg lamps. PMID- 11478248 TI - Formation and emission of volatile polonium compound by microbial activity and polonium methylation with methylcobalamin. AB - We observed biologically mediated emission of Po from culture solution inoculated sea sediment extract and incubated under natural light/dark cycle condition or dark condition the emitted Po compound would be lipophilic because of effective collection in organic solvent. Sterilization of the culture medium with antibiotics or CuSO4 completely suppressed growth of microorganisms and resulted in no emission of Po, indicating biological activity of microorganisms is responsible for formation and emission of volatile Po compound. Po emission also occurred when seawater was used as a culture medium. Our finding indicates a possibility of biotic source for atmospheric Po in the environment, which has been believed to be originated from abiotic sources. We compared emission behavior of Po and S in the culture experiments, the elements belong to XVI group in the Periodical Table, and consider that their emission mechanisms involved would be different though the emission of both elements is supported by biological activity of microorganisms. One of the chemical forms of S emitted was confirmed to be dimethyl sulfide (DMS) but that of Po is not known. Methylation experiments of Po with methylcobalamin demonstrated a formation and emission of volatile Po compound. The methylation of Po with methylcobalamin might be related to the observed Po emission in the culture experiments. PMID- 11478249 TI - Evidence for simultaneous abiotic-biotic oxidations in a microbial-Fenton's system. AB - The conditions that support the simultaneous activity of hydroxyl radicals (OH.) and heterotrophic aerobic bacterial metabolism were investigated using two probe compounds: (1) tetrachloroethene (PCE) for the detection of OH. generated by an iron-nitrilotriacetic acid (Fe-NTA) catalyzed Fenton-like reaction and (2) oxalate (OA) for the detection of heterotrophic metabolism of Xanthobacter flavus. In the absence of the bacterium in the quasi-steady-state Fenton's system, only PCE oxidation was observed; conversely, only OA assimilation was found in non-Fenton's systems containing X. flavus. In combined Fenton's microbial systems, loss of both probes was observed. PCE oxidation increased and heterotrophic assimilation of OA declined as a function of an increase in the quasi-steady-state H2O2 concentration. Central composite rotatable experimental designs were used to determine the conditions that provide maximum simultaneous abiotic-biotic oxidations, which were achieved with a biomass level of 10(9) CFU/mL, 4.5 mM H2O2, and 2.5 mM Fe-NTA. These results demonstrate that heterotrophic bacterial metabolism can occur in the presence of hydroxyl radicals. Such simultaneous abiotic-biotic oxidations may exist when H2O2 is injected into the subsurface as a microbial oxygen source or as a source of chemical oxidants. In addition, hybrid abiotic-biotic systems could be used for the treatment of waters containing biorefractory organic contaminants present in recycle water, cooling water, or industrial waste streams. PMID- 11478250 TI - Reactivity of Pb(II) at the Mn(III,IV) (oxyhydr)oxide--water interface. AB - In this study, the reactivity of lead (Pb(II)) on naturally occurring Mn(III,IV) (oxyhydr)oxide minerals was evaluated using kinetic, thermodynamic, and spectroscopic investigations. Aqueous Pb(II) was more strongly adsorbed to birnessite (delta-MnO1.7) than to manganite (gamma-MnOOH) under all experimental conditions. The isoteric heat of Pb adsorption (delta HT) or birnessite was 94 kJ mol-1 at a surface loading of 1.1 mmol g-1, and decreased with increasing adsorption density. This indicated that adsorption was an endothermic process and that birnessite possessed heterogeneous sites of reactivity for Pb. X-ray absorption fine structure (XAFS) spectra revealed that Pb was adsorbed as inner sphere complexes on both birnessite and manganite with no evidence to suggest oxidation as an operative sorption mechanism. Lead appeared to coordinate to vacancy sites in the birnessite layer structure with concurrent release of Mn to solution, which resulted in a greater number of second shell Mn scatterers in Pb birnessite when compared to Pb-manganite samples. The difference in Pb coordination apparently explained the contrasting desorption behavior between the two Mn minerals. These results have significant implications for Pb partitioning in soil environments containing solid-phase Mn(III,IV) (oxyhydr)oxides. PMID- 11478251 TI - Solid-state 2H MAS NMR studies of TNT absorption in soil and clays. AB - The binding of TNT to a soil and soil components is studied with 2H MAS NMR. This NMR technique clearly shows different interactions for deuterated TNT with soil, quartz, and clays. Basically, this NMR method relies on the well-defined 2H MAS NMR characteristics of a deuterated methyl group attached to an aromatic ring. Stationary rings and rings executing 2- or 3-fold jump motions all yield different NMR spectra. The technique has good sensitivity; these studies were conducted at a TNT loading of 22 mmol/kg. For the soil sample studied, TNT showed binding similar to a TNT/K10-montmorillonite sample. PMID- 11478252 TI - Sensitivity of particulate matter nitrate formation to precursor emissions in the California San Joaquin Valley. AB - The formation of secondary ammonium nitrate during the 1995 Integrated Monitoring Study (IMS95) in San Joaquin Valley, CA was investigated using a box model that simulates the atmospheric chemistry and gas/particle partition of inorganic compounds. The concentration of particulate matter (PM) nitrate was found to be sensitive to reductions in VOC emissions. Nitric acid, rather than ammonia, was the limiting reagent in the formation of PM nitrate. The formation of nitric acid was more sensitive to the availability of oxidants than that of NOx. Oxidant chemistry in wintertime conditions in the San Joaquin Valley was shown to be VOC sensitive. In fact, a decrease in NOx emissions may have the counter-intuitive effect of increasing PM nitrate. PMID- 11478253 TI - Transparent thin-film TiO2 photocatalysts with high activity. AB - A transparent, thin-film TiO2 layer prepared by sol-gel deposition is shown to be more active for photocatalytic oxidation (PCO) of acetaldehyde, acetic acid, and toluene than Degussa P25 thin films. The sol-gel TiO2 adsorbs 30-70% less organic, but the PCO activity per adsorbed molecule is 3.5-8.5 times higher on the sol-gel TiO2 than on Degussa P25. In addition, less-reactive intermediates do not appear to form as readily on the sol-gel catalyst as they do on Degussa P25, and thus the sol-gel catalyst deactivates slower during toluene PCO. Rates were measured in transient experiments for a monolayer of adsorbed organic, and transient experiments are shown to be an effective way to measure rates, selectivities, surface coverages, and formation of less-reactive intermediates in the same set of experiments. PMID- 11478254 TI - Solvent extraction for separating micellar-solubilized contaminants and anionic surfactants. AB - Decontamination of contaminant-laden surfactant solutions is critical to successful implementation of surfactant-enhanced aquifer remediation (SEAR). Solvent extraction was studied for removing micellar-solubilized contaminants having low equivalent alkyl carbon numbers (EACNs) from surfactant solutions. Factors influencing the solvent extraction of micellar-solubilized contaminant were studied, including surfactant concentration, solution salinity, solvent solubilization, and solvent/solution volumetric ratio. A model was developed to quantify the impacts of these factors on contaminant removal. The good agreement between experimental results and model predictions corroborates assumptions made in the model development. From these results, it is concluded that extracting solvents must have much higher EACNs than that of the contaminant to reduce the micellar solubilization of the solvents, which can significantly reduce contaminant removal efficiency. However, the highest EACN solvent is not necessarily the best one for contaminant removal due to other constraints (e.g., molecular weight and viscosity). Increasing the total surfactant concentration or salinity of an anionic surfactant solution increases its contaminant solubilization capacity but reduces the contaminant removal efficiency by solvent extraction. Continuous column extraction operated at a low column surface loading rate allowed contaminant partitioning to approach equilibrium conditions, and multistage column extraction was able to improve the contaminant removal efficiency while minimizing solvent requirement. PMID- 11478255 TI - Dioxin and furan formation in FBC boilers. AB - Fluidized bed combustion (FBC) is increasingly being used for municipal solid waste incineration and for various applications in which dioxin and furan (PCDD/F) formation are concerns. Despite the fact that fluidized bed combustion is a relatively low temperature system, current evidence shows that they perform relatively well. One hypothesis is that sulfur, which is contained in some of the fuels used in FBC (e.g. coal in cofiring situations), helps to reduce dioxin production. This paper endeavors to examine this speculation and finds that such benefits depend on the CI/S ratio and that at higher sulfur addition levels dioxins, furans, and polycyclic aromatic hydrocarbon (PAH) production may actually increase. This and the reasons for it are discussed. PMID- 11478256 TI - Membrane rejection of nitrogen compounds. AB - Rejection characteristics of nitrogen compounds were examined for reverse osmosis, nanofiltration, and low-pressure reverse osmosis membranes. The rejection of nitrogen compounds is explained by integrating experimental results with calculations using the extended Nernst-Planck model coupled with a steric hindrance model. The molecular weight and chemical structure of nitrogen compounds appear to be less important in determining rejection than electrostatic properties. The rejection is greatest when the Donnan potential exceeds 0.05 V or when the ratio of the solute radius to the pore radius is greater than 0.8. The transport of solute in the pore is dominated by diffusion, although convective transport is significant for organic nitrogen compounds. Electromigration contributes negligibly to the overall solute transport in the membrane. Urea, a small organic compound, has lower rejection than ionic compounds such as ammonium, nitrate, and nitrite, indicating the critical role of electrostatic interaction in rejection. This suggests that better treatment efficiency for organic nitrogen compounds can be obtained after ammonification of urea. PMID- 11478257 TI - Applications of ultrasound in NAPL remediation: sonochemical degradation of TCE in aqueous surfactant solutions. AB - Surfactant-enhanced pump-and-treat technologies increase the efficiency of nonaqueous-phase liquids (NAPLs) removal from soils. However, high concentrations of surfactants in groundwater impose severe limitations to water treatment. In this paper, we explore the applicability of ultrasonic irradiation as an alternative method for surfactant recovery and contaminant degradation. The combined effects of temperature, initial substrate concentration, and concentration of added surfactant (sodium dodecyl sulfate, SDS) were analyzed for the sonolysis of trichloroethylene (TCE) in batch experiments at an ultrasonic frequency of 500 kHz and 77 W/L applied power density. In the range of 5-30 degrees C, TCE sonolysis becomes faster at higher temperatures, both in the absence and in the presence of surfactant. This indicates that gas-phase pyrolysis prevails over other chemical reactions in the liquid phase. Inhibition of TCE sonolysis was observed in the presence of surfactant at all SDS concentrations. Changes in the initial TCE concentration (from 250 microM to 1.2 mM) showed no effect on the degradation rates in the presence of SDS. For surfactant levels below its critical micelle concentration (cmc), the inhibition of TCE sonolysis exhibited a highly nonlinear dependence with increasing SDS concentration. A correlation was observed in this range between the relative inhibition of sonolysis and the decreasing surface tension of the solutions. Above the cmc up to an SDS concentration of 5%, the reaction rate decreased less markedly. Micellar sequestration of the contaminant seems to be the main reason for this additional inhibition. Bubble growth prior to collapse may incorporate some of the TCE dissolved in the micelles through their adsorption in the expanding bubble walls, thus partially overcoming the scavenging effect due to micellar entrapment of the contaminant. PMID- 11478258 TI - Study of the mechanisms of cadmium biosorption by dealginated seaweed waste. AB - The ability of dealginated seaweed waste, a waste material derived from the commercial processing of seaweed for alginate production, to remove cadmium from solution was determined. Cadmium sorption was found to be rapid (91% removal within 5 min), achieving a residual concentration of 0.8 mg L-1 after 1-h contact time from an initial solution concentration of 10 mg L-1. The binding of cadmium by dealginate was found to be pH dependent, optimal sorption occurring at around pH 6-8. The mechanism of cadmium ion binding by dealginate was investigated by a number of techniques. Potentiometric titration of the dealginate revealed two distinct pKa values, the first having a value similar to carboxyl groups and the second comparable with that of saturated thiols and amines. Esterification of the dealginate resulted in the subsequent reduction in cadmium sorption (95% to 17%), indicating that carboxyl groups are largely responsible for sorption. Evidence from FT-IR spectra confirmed the presence of carboxyl groups in untreated dealginate, while the number of carboxyl groups was markedly reduced in the esterified sample. Furthermore, the FT-IR spectrum for dealginate was found to be similar to that previously reported for mannuronic acid-rich calcium alginate. Determination of a molar ratio in the displacement of calcium by cadmium on dealginate further supported the presence of an ion-exchange relationship. The ion-exchange constant was calculated to be 0.329 x 10(-6). The speciation of cadmium in solution both before and after sorption was determined by an ion selective electrode (ISE) technique. The findings of this study suggest that the sorption of cadmium by dealginate is mainly due to an ion-exchange mechanism. PMID- 11478259 TI - Remediation of DNAPL pools using dense brine barrier strategies. AB - Although dense nonaqueous phase liquid (DNAPL) pools are an important source of groundwater contamination, little experimental data have been generated to develop a mature level of understanding of the problem, and few strategies specifically aimed at remediation have been advanced. We discuss the dominant importance of these features in subsurface systems, present novel two- and three dimensional heterogeneous experimental systems, and show results from two evolving strategies for remediating DNAPL pools. These strategies involve the joint use of a dense brine barrier and controlled mobilization of trapped DNAPL using small-volume surfactant flushes. These experiments demonstrate a controlled, substantial reduction of entrapped DNAPL in both two- and three dimensional heterogeneous domains, using less than a single pore volume of flushing solution in some cases. PMID- 11478260 TI - Studies on the interfacial characterization of O/W emulsion for the optimization of its treatment. AB - Wastewater produced in the form of an o/w emulsion is difficult to treat properly because of its complex behavior. To acquire information for the optimization of emulsion controlling processes, the interfacial features of emulsion systems have been investigated in this work. The surface tension is found to decrease as the oil content is increased, and contact angle is influenced by the hydrophobicity of the substrate. The work of adhesion for substrates with different degrees of hydrophobicity has been estimated from the contact angle and surface tension. Temperature and pH have a significant influence on the turbidity and surface tension, and it is observed that electrophoretic characteristics of oil droplets are closely related with turbidity. Also, effects of inorganic salts and surfactants on the electrokinetic properties of emulsions have been considered, and the influence of coagulant on the stability is discussed through the calculation of potential energy of interaction between oil droplets. Adsorption and electrocoagulation processes have been attempted under various conditions in order to examine the practical treatment efficiencies of emulsion in relation to its interfacial characteristics. PMID- 11478261 TI - Modeling of copper biosorption by Arthrobacter sp. in a UF/MF membrane reactor. AB - Copper biosorption by Arthrobacter sp. has been studied in this work. The process has been realized inside of a ultrafiltration/microfiltration (UF/MF) reactor in order to confine cells. A mathematical model has been developed that is able to predict experimental data under different operating conditions. The model takes into account different phenomena, which might occur during the process, such as a dependence of equilibrium parameters on pH, a partial cell disruption, and a change in the membrane retention properties at high biomass concentrations. Experimental tests have been performed under different operating conditions: a full factorial design has been implemented with pH (levels: 4, 5, and 6 units) and biomass concentration (levels: 1 and 5 g/L) as factors. A simple mathematical model based on metal mass balance taking into account the effect of pH on the Langmuir equilibrium adsorption parameters well fitted experimental data at low pH values and biomass concentrations. A more complex mathematical model, which considers a partial cell disruption during the biosorption trial, was proposed to understand and analyze the anomalous system behavior at pH = 6 and biomass concentration equal to 5 g/L. The effect of mechanical stress on biomass performances was also examined by using a discontinuous system (test tube trials) simulating the membrane reactor apparatus. In this alternative system biosorption trials were carried out in test tubes in such a way to avoid or at least minimize the disruption due to mechanical stress. Experimental results obtained by using this system can be modeled up to pH = 5 without considering cell disruption phenomenon, while at pH = 6 possible chemical reactions of biomass constituents could happen. PMID- 11478262 TI - Release of CFC-11 from disposal of polyurethane foam waste. AB - The halocarbon CFC-11 has extensively been used as a blowing agent for polyurethane (PUR) insulation foams in home appliances and for residential and industrial construction. Release of CFCs is an important factor in the depletion of the ozone layer. For CFC-11 the future atmospheric concentrations will mainly depend on the continued release from PUR foams. Little is known about rates and time frames of the CFC release from foams especially after treatment and disposal of foam containing waste products. The CFC release is mainly controlled by slow diffusion out through the PUR. From the literature and by reevaluation of an old reported experiment, diffusion coefficients in the range of 0.05-1.7 x 10(-14) m2 s-1 were found reflecting differences in foam properties and experimental designs. Laboratory experiments studying the distribution of CFC in the foam and the short-term releases after shredding showed that about 40% of the CFC is solubilized in the PUR phase, and that up to 10% of the total content will be released within a few weeks if the foam is shredded down to 2-cm sized pieces. For smaller pieces the quick release will be larger. Fifty percent of residual CFC content will be released within 9-300 years from 2-cm pieces based on the range in diffusion coefficients reported. For larger pieces the initial release is insignificant, and the release time frames are much longer than for the shredded foam. PMID- 11478263 TI - The cadherin-catenin superfamily in endocrine tumors. AB - It has been well-known that the cadherin-catenin complexes bind with intracellular skeleton actin, which result in stabilization of cellular structure and tissue organization. Therefore, the cadherin-catenin family has been considered prerequisite for normal cell function and the preservation of tissue integrity. In human malignancies especially colon cancers, dysfunction and/or decrease of expression of these proteins have been proposed to prevent differentiation of tumors and to increase invasiveness and poor prognosis. However, recent studies also revealed that a member of this superfamily, beta catenin, may play an important role in Wnt/wingless intracellular signaling pathway. Decreased expression of this protein or somatic mutation of the beta catenin gene has been also reported in human carcinomas including various endocrine tumors. Mutant beta-catenin is associated with abnormal nuclear accumulation in tumor cells and subsequently to activate other transcription factors such as Tcf/Lef. This activation eventually results in which upregulation of mRNA and protein levels of various cell growth mediators in these endocrine tumors. Therefore, dysfunction of the cadherin-catenin system is considered to be closely correlated with tumorigenesis and development in human endocrine tumors. PMID- 11478264 TI - Expression of sodium iodide symporter in benign and malignant human thyroid tissues. AB - The extent of human sodium iodide symporter (hNIS) expression in different kinds of human thyroid cancer tissues and cell lines remains controversial. In this study, polyclonal antibodies to hNIS were used to analyze the expression of symporter protein in benign and malignant human thyroid tissues. Formalin-fixed, paraffin wax-embedded tissue sections were used. Staining was performed using primary polyclonal antibody of rabbit anti-human hNIS diluted in PBS (1:500). Results showed that 2 of 3 normal tissue, 3 of 6 nodular hyperplasia, one follicular adenoma, 3 of 11 papillary thyroid carcinoma, 1 of 5 follicular carcinoma and none of 3 metastatic thyroid epithelial tissue specimens stained positively for hNIS. A higher percentage of positive staining for symporter protein was found in benign thyroid tissues including normal thyroid tissue, nodular hyperplasia, and adenoma (60%). In contrast, papillary and follicular thyroid carcinomas demonstrated lower symporter protein expression (20%). In conclusion, although the number of tissue samples examined in this study was small, hNIS staining found a higher ratio of symporter protein expression in normal and benign thyroid tissues compared with malignant tissues. Determination of the reason for discrepancies in the expression of hNIS in in vivo and in vitro studies will require further investigation. PMID- 11478265 TI - Latent carcinoma of the thyroid in Austria: a systematic autopsy study. AB - Data concerning the incidence of latent thyroid carcinoma (LTC) in populations with endemic goiter are scarce. Despite the introduction of iodine goiter prophylaxis in the early sixties, iodine supply is still insufficient in Austria and goiter remains endemic. This is the first detailed study dealing with epidemiological features of LTC at autopsy in Austria. A total of 118 thyroid glands were included in the study. The glands were serially sectioned at 2- to 3 mm intervals, embedded in paraffin and histologically examined for the presence of LTC. In addition, the incidence and severity of lymphocytic thyroiditis (LT) were evaluated. Ten thyroids were found to contain LTC (8.6%). All were of the papillary type. The mean tumor dimension was 4.9 mm +/- 3.2, the smallest lesion measuring 1 mm. Only the largest tumor slightly exceeded the extent of a microcarcinoma and measured 10.5 mm. Multifocal disease was present in three cases (30%). The prevalence of latent papillary thyroid carcinoma (LPTC) was 6.6% (n = 4) in females and 10.5% (n = 6) in males. The mean age of the subjects with LPTC was 67.7 +/- 14.4 yr, range 37 to 77 yr. Goitrous thyroids were seen in 33 cases (28%): One gland was diffusely enlarged and 32 (27.1%) enlarged glands were nodular goiters. The overall prevalence of LT was 30.5% (n = 36) and the only type of thyroiditis observed was focal lymphocytic thyroiditis (FLT). There was no correlation between the presence of LPTC and goiter, the presence of FLT and the subjects' age and sex. The incidence of LPTC in Austria is similar to that in nongoitrous regions. The adult population at large seems to be uniformly exposed to factors involved in the initiation and early growth of papillary thyroid carcinoma (PTC). This suggests that the levels of iodine intake only play a minor role in the early phase of the carcinogenesis of PTC, but may be of some importance in the progression of LPTC to clinically evident PTC. PMID- 11478266 TI - Morphological analysis of adrenal glands: a prospective analysis. AB - There is a lack of studies to document the weight range of normal adrenal glands. The aims of the current study are to find out the weight range of normal adrenal glands in Chinese patients and to analyze any potential factors affecting it. Adrenal glands not affected by disease were prospectively collected from autopsies on 333 Chinese patients (208 men, 125 women). The weight and longest dimension of each adrenal gland were noted. The impact of various clinicopathological factors on the adrenal weight was studied. In addition, the morphometric features of these adrenal glands were compared with that of 28 surgically resected adrenal glands with cortical adenomas. The mean combined weight of adrenal glands was 11.8 g (range = 5.8 g to 19.9 g). The left adrenal gland was often heavier than the right (mean weight = 6.1 g and 5.7 g respectively) and with a greater longitudinal length than the right (mean length = 5.2 cm and 4.8 cm respectively). Adrenal glands were often heavier in male, younger age group (less than 60-yr-old) and patients with history of hypertension or lung cancers. In comparison, the mean weight of right adrenal gland with cortical adenoma was 11.7 g (range = 6 g to 26 g) and that of left adrenal gland with cortical adenoma was 9.4 g (range = 4.6 g to 25 g). This is the first study in the English literature that provides data on the weight of normal adrenal glands in a solely Chinese population. Adrenal weight may be affected by patients' gender, age, laterality of adrenal gland and presence of systemic disease. PMID- 11478267 TI - Cyclins D1 and D3 and topoisomerase II alpha in inactive pituitary adenomas. AB - The oncogenes cyclin D1 and D3 are overexpressed in many tumors. Topoisomerase II alpha is found in proliferating cells. The immunohistological expression of cyclin D1, cyclin D3, and Topoisomerase II alpha was studied in a collection of 60 clinically inactive surgically removed pituitary adenomas of the follicle stimulating hormone/luteinizing hormone (FSH/LH) cell complex (20 null cell adenomas, 20 oncocytomas, and 20 FSH/LH cell adenomas) for correlation with other proliferation markers (Ki-67, PCNA) and with clinical data. Whereas cyclin D1 was positive only in one invasive null cell adenoma (1.7%) with some p53-positive nuclei, cyclin D3 was overexpressed in the nuclei of 41 tumors (68%). Topoisomerase II alpha was demonstrated in the nuclei of 42 adenomas (70%) with no significant differences discernible between the three adenoma subtypes. There was no significant correlation to the time of development of tumor symptoms, but a correlation of Topoisomerase II alpha with cyclin D3 and the proliferation marker Ki-67 (Mib1). From these data we conclude that cyclin D3 and Topoisomerase II alpha appear to be additional markers for proliferation which can be used for prognosis index in surgical pathology of the pituitary. PMID- 11478268 TI - Histologic diagnosis of primary hyperparathyroidism: a concordance analysis between three pathologists. AB - Primary hyperparathyroidism (HPT) is caused by a parathyroid adenoma, hyperplasia or carcinoma. Difficulties for the histologic diagnosis of abnormal parathyroid tissue are widely recognized. The aim of the study was to evaluate the reproducibility of the morphologic criteria through a concordance study among three pathologists. Representative slides of 40 patients with biochemically primary HPT stained with hematoxylin and eosin were blindly reviewed by three pathologists. Each pathologist established the diagnosis of adenoma or hyperplasia and assessed the presence of fat cells, a rim of normal tissue, a fibrous capsule, the number of cellular types, the lobular pattern, and the characteristics of the blood vessel's wall. A concordance analysis was then performed. Mean age of the group was 55 +/- 14 yr, 7 were males and 33 females. The concordance analysis among the three pathologists for the differential diagnosis between adenoma and hyperplasia, showed a Kappa index of 0.5. Kappa index for the presence of fat cells was 0.56, for the presence of a rim of normal tissue 0.47, and for the number of cellular types 0.29. The concordance for the differential diagnosis between parathyroid adenoma and hyperplasia in this study was low. PMID- 11478269 TI - Clinicopathological value of somatostatin type 2A and estrogen receptor immunoreactivity in human breast carcinoma. AB - Somatostatin type 2A (sstr2A) and estrogen receptor (ER) are interrelated regulatory receptors present in normal breast epithelium and in a population of breast carcinomas. ER mediates growth stimulatory effects of estrogens whereas sstr2A mediates growth inhibitory actions of somatostatin. However, much work has been devoted to elucidate the biological role of ER, little is known about sstr2A in breast cancer. In the present study we examined immunoreactivity of sstr2A and ER in 64 breast carcinomas in correlation with tumor size and histological grade (HG), presence of lymph node metastasis (LNM), Nottingham prognostic index (NPI), and the patients' age. ER and sstr2A immunoreactivity were present in 78% and 63% of the breast carcinomas, respectively. Ninety percent of tumors immunoreactive for sstr2A were simultaneously immunoreactive for ER. ER immunoreactivity correlated significantly with lower HG (p = 0.03) and better NPI (p = 0.02). sstr2A immunoreactivity correlated significantly with lower HG (p = 0.012) but not with NPI (p = 0.26). There was no correlation of sstr2A immunoreactivity and tumor size, patients' chronological age or LNM. The results confirm prognostic value of ER immunohistochemistry in breast carcinoma. However sstr2A cannot substitute ER for prognostic evaluation, sstr2A immunoreactivity being significantly associated with lower HG seems to represent an independent prognostic factor in breast carcinoma. PMID- 11478270 TI - Cysts in the rat adenohypophysis: incidence and histology. AB - The incidence and histology of cysts in the adenohypophysis of adult male Wistar rats are reported. Of sixty pituitaries studied 13 of them (21.6%) presented a single cyst located in the pars distalis. The cysts varied in shape and size and were usually multilocular. Two of them were connected with the subdural space at the ventral surface of the adenohypophysis. Histology demonstrated that the cysts were filled with mucinous material and foamy macrophages and were lined by flat and cuboidal ciliated and nonciliated epithelial cells, goblet cells as well as several adenohypophysial endocrine cells such as somatotrophs, thyrotrophs, and gonadotrophs. The ciliated cells were the most numerous. Histologic and immunohistochemical studies of the uninvolved areas of the adenohypophysis showed no abnormalities and the weights and histology of the adenohypophyses and peripheral endocrine glands were within normal range, suggesting that the cysts did not impair the adenohypophysial endocrine activity. Although the morphogenesis of the cysts remained obscure, the histological and immunohistochemical findings support the hypothesis that during embryonic development, the future cysts coming from the pharyngeal epithelium is fused with the stomodeum before or during the formation of the Rathke's pouch. PMID- 11478271 TI - Papillary carcinoma occurring within an adenomatous goiter of the thyroid gland in Cowden's disease. AB - Cowden's disease is an autosomal dominant disorder characterized by multiple benign and malignant neoplastic lesions involving many organs. The presence of characteristic cutaneous lesions is crucial for the diagnosis. Thyroid disease is a major extracutaneous manifestation of this disease; however, the histologic characteristics have not been described in detail. We report a case of thyroid tumor associated with Cowden's disease. Grossly, the tumor showed a multinodular appearance, like an adenomatous goiter. Microscopically, it consisted of follicular adenomas with a trabecular pattern. Some of the nodules had a second component resembling papillary carcinoma. This was thought to be a unique histological feature not described previously, and might be specific to thyroid tumor associated with Cowden's disease. PMID- 11478272 TI - Histological, immunohistochemical, ultrastructural and biochemical study of human gastric composite tumor: expression of the serotonin-2B receptor by the neuroendocrine component. AB - We report a case of a human gastric composite tumor occurring seven years after a partial gastrectomy for a low grade B cell MALT lymphoma. Histological examination of the tumor revealed two intimately intermingled components: 1. A moderately to poorly differentiated tubulo-acinar adenocarcinoma with signet-ring cells; and 2. Isolated or clustered small neuroendocrine cells without atypia expressing chromogranin A, somatostatin and/or glucagon, serotonin (5-HT) and, the 5-HT2B receptors. In addition to immunohistochemical detection, the presence of 5-HT2B receptors was shown pharmacologically through [125I]-DOI binding. Since 5-HT2B receptors have been demonstrated to have autocrine functions and, mitogenic and transforming properties, these results suggest a role of 5-HT in neuroendocrine malignant transformation. On the other hand, the expression of somatostatin and the detection by reverse transcriptase polymerase chain reaction (RT-PCR) of somatostatin receptor subtypes 2, 3, and 5, which have been shown to be involved in tumor regression, might account for the long evolution of this case (> 5 yr). This case illustrates the importance of local humoral modulation in tumor growth. Moreover, ultrastructural results favor a unique origin of the tumor cells from one amphicrine cell type. PMID- 11478273 TI - Isolated Pneumocystis carinii infection of adrenal glands causing Addison's disease in a non-immunocompromised adult. AB - Pneumocystis carinii is primarily an opportunistic pathogen infecting patients with AIDS and other immunocompromised patients, and ordinarily does not affect immunocompetent persons. We report isolated P. carinii infection of bilateral adrenal glands in a non-immunocompromised adult male, leading to fatal Addisonian crisis. Diagnosis of P. carinii was established on the basis of cytopathology and microbiological tests, using conventional staining techniques and direct immunofluorescence on ultrasound-guided fine needle aspirates and trucut needle biopsy specimen from adrenal glands. P. carinii pneumonia and other fungal infections of the adrenal glands were excluded by appropriate tests. Absence of HIV infection was established by negative ELISA for HIV I and II antibodies and Western blot analysis at the time of presentation and 45 d later. Normal blood total leukocyte and CD4 lymphocyte counts and IgG and IgA levels confirmed the immunocompetent status of the patient. The patient improved with anti Pneumocystis treatment and corticosteroid replacement, but succumbed to an episode of Addisonian crisis triggered by a diarrheal illness. PMID- 11478274 TI - [Effect of the preparation Wobe-Mugos E on parameters of the antioxidant defence system and morphofunctional erythrocyte status in patients with chronic obstructive bronchitis]. AB - In the examination of 64 patients with chronic obstructive bronchitis, some specificities of functioning of decompensation mechanisms in the glutathione antiradical system were established as were changes in morphofunctional and receptor properties of erythrocytes in chronic obstructive bronchitis. The patients were shown to have derived benefit from a combined treatment involving the use of the enzymic drug preparation Wobe-Mugos E which was found to make for improvement of rheological properties of erythrocytes, their capability of depositing and transporting catecholamines. PMID- 11478276 TI - Bibliography. Current world literature. Molecular genetics. PMID- 11478275 TI - Special issue in memory of Professor Clement Henri Bamford. PMID- 11478277 TI - Bibliography. Current world literature. Cardiac failure. PMID- 11478278 TI - Reporting net moments about the lumbar spine [letter]. PMID- 11478279 TI - Meeting hints at thaw in relations between genome rivals. PMID- 11478280 TI - [Biological rhythm and man's environment: work and jet lag]. PMID- 11478281 TI - [Biological rhythms and human disease]. PMID- 11478282 TI - Treatment tools in chronobiology. PMID- 11478283 TI - [What strategy is best for suspected toxic drug dermatitis in patients taking several drugs?]. PMID- 11478285 TI - [Quid? Ovarian granulosa tumor manifesting as pseudo-precocious puberty]. PMID- 11478286 TI - [Quid? Cystic compression of the right suprascapular nerve in the spinoglenoid notch]. PMID- 11478287 TI - Bibliography. Current world literature: diseases of the pleura. PMID- 11478288 TI - Bibliography. Current world literature: neoplasms of the lung. PMID- 11478289 TI - The test distribution of modularity statistics: a correction and clarification. PMID- 11478290 TI - Improving the estimation of bacterial allele frequencies. PMID- 11478291 TI - Forced titration as a confound in an SSRI comparator study of sertaline versus citalopram in the treatment of major depressive disorder. PMID- 11478292 TI - Hippocampal volume and cognition in geriatric depression. PMID- 11478293 TI - Symposium proceedings: Pharmacia Diagnostics, Stallergenes, Hitachi Diagnostics. PMID- 11478294 TI - [Psychological aspects of emotional regulation in smoking dependency]. AB - We reviewed the literature on the relationships between smoking and affect regulation. There is strong evidence that vulnerability to smoking dependence is a function of a high initial sensitivity to nicotine, which produces reinforcing consequences that lead to chronic use. These strong reinforcers of tobacco dependence include regulation of mood and modulation of arousal. We focused on studies concerned with the subjective component of arousal and emotional experience. We discuss first the models and classifications used to differentiate types of smoking as related to the management of emotions and studies evaluating the stimulant and subjective effects of smoking behavior, questioning the paradoxical tranquilizing and anxiety-reducing effects of nicotine. We also looked into the mood regulation effects that may explain the strong relationships observed between depression and smoking and finally focus on some of the personality risk factors that may make individuals more susceptible to these rewarding properties of smoking. PMID- 11478295 TI - [Attachments and addictive behaviors]. PMID- 11478296 TI - A trachoma perspective. PMID- 11478297 TI - [Does the medical informatics program only serve to classify hospitals?]. PMID- 11478298 TI - [Does the medical informatics program only serve to classify hospitals?]. PMID- 11478299 TI - [Clinical and economic evaluation of laparoscopic surgery of groin hernia]. PMID- 11478300 TI - Near haploid blast phase in a chronic myeloid leukemia detected by fluorescence in situ hybridization using a BCR-ABL probe. PMID- 11478301 TI - Childhood acute lymphoblastic leukemia with a novel der(10)t(7;10)(q11;q26). PMID- 11478302 TI - Constitutional t(5;11)(p15.3;q23) in an adolescent male with acute lymphoblastic leukemia. PMID- 11478303 TI - Does the PITSLRE gene complex contribute to the pathogenesis of malignant melanoma of the skin? A study of patient-derived tumor samples? PMID- 11478304 TI - VIIth Annual Congress of the Center for Studies of Human Polymorphism. Paris, France, 24-26 May 2000. Proceedings and abstracts. PMID- 11478306 TI - First report of AIDS. PMID- 11478305 TI - VIII International Congress of Reproductive Immunology. Opatija, Croatia, July 2 6, 2001. Abstracts. PMID- 11478307 TI - [Neurology meetings of the French-speaking community. Lyon, France 18-21 April 2001. Abstracts]. PMID- 11478308 TI - European Federation of National Associations of Orthopaedics and Traumatology. Rhodes, Greece, June 1-7, 2001. Abstracts. PMID- 11478309 TI - International Congress of Sexually Transmitted Infections. 24-27 June 2001, Berlin, Germany. Abstracts. PMID- 11478310 TI - British Cancer Research Meeting 2001. Leeds, United Kingdom, 1-4 July 2001. Abstracts. PMID- 11478311 TI - Carcinogenesis testing updated. PMID- 11478312 TI - Carcinogen induced unscheduled DNA synthesis in mouse hepatocytes. AB - Mouse primary liver cell cultures were examined for evidence of unscheduled DNA synthesis (UDS) following treatment with the carcinogens; dimethylnitrosamine (DMNA), diethylnitrosamine (DENA), 2-acetylaminofluorene (2-AAF), N-methyl-N' nitro-N-nitrosoguanidine (MNNG), benzo(a)pyrene (BP), dimethylbenzanthracene (DMBA), 1,1,-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT), safrole, diethylstilbestrol (DES), aflatoxin B1 (AFB1), and dieldrin and the noncarcinogens; dimethylformamide (DMF), fluorene, and pyrene. Mouse hepatocyte cultures were simultaneously treated with three concentrations of each compound and 3H-thymidine. After 24 hrs, cells were fixed and processed for autoradiography. 3H-thymidine incorporation in both experimental and control cell nuclei, as evidenced by autoradiographic grains, was quantitated microscopically. DMNA, DENA, 2-AAF, MNNG, BP, AFB1 and DMBA significantly increased UDS over untreated cells at all concentrations studied. DDT, DMF, fluorene, pyrene, safrole, DES, and dieldrin were negative for UDS in all concentrations examined. DMNA, 2-AAF and MNNG were also studied for UDS induction in 2 hr old, 1 day old and 4 day old cultures. A progressive decrease in UDS with increased time after plating was found in DMNA and 2-AAF treated cultures. After 4 days DMNA and 2-AAF induced UDS only at the highest concentrations examined (10(-3) M and 10(-4) M respectively). MNNG induced UDS at all time periods and concentrations sampled. An attempt to enhance the sensitivity of the UDS assay by inducing the mixed function oxidative enzyme activity in the hepatocytes with phenobarbital administered in vivo resulted in no statistically significant increase in UDS with DMNA, 2-AAF, MNNG, DDT, and dieldrin when compared with cells from non induced animals. PMID- 11478313 TI - Use of historical control data in carcinogenicity studies in rodents. AB - This paper considers the use of historical control data in the evaluation of tumor incidences from carcinogenicity studies in rodents. Although the most appropriate control group for interpretative purposes is always the concurrent control, there are instances in which the use of historical control information can aid an investigator in the overall evaluation of tumor incidence data. One example is rare tumors; another is a tumor that shows a marginally significant result relative to concurrent controls. However, before historical control data can be used in a formal testing framework, a number of important issues must first be considered. The nomenclature conventions and diagnostic criteria for each study should be identical to insure unambiguous identification of all relevant tumors in the historical control database. Criteria should be established that will aid in determining whether a particular study should be included in the database. This will assure a homogeneous set of studies upon which to base statistical comparisons. Since study-to-study variability in tumor rates may exceed what would be expected by chance alone, these sources of variability should be identified and controlled. Finally, statistical procedures should be employed that adjust for extra-binomial variability. This paper also summarizes tumor incidence data from untreated Fischer 344 rats and B6C3F1 mice in the National Toxicology Program (NTP) historical control database. All studies in the database are of two years duration, and all neoplasms occurring with a frequency of 0.5% or more are reported. PMID- 11478314 TI - Abnormalities in liver iron accumulation during N-2-fluorenylacetamide hepatocarcinogenesis that are dependent or independent of continued carcinogen action. AB - The characteristics of liver iron accumulation were studied during N-2 fluorenylacetamide (FAA)-induced hepatocarcinogenesis in rats. After injection of iron-dextran in control rats, hepatocytes accumulated stainable iron evenly throughout hepatic lobules. During the feeding of FAA, iron accumulation was reduced in the midzonal and centrilobular regions. After FAA removal, hepatocytes in these regions again accumulated high amounts of iron. Hepatocellular altered foci induced by FAA displayed rather uniform (> 94%) iron-exclusion during FAA feeding. After FAA removal, however, iron-exclusion was lost in a fraction of the foci, while others (40-64%) remained resistant to iron accumulation. A large majority of liver neoplasms (> 93%) displayed resistance to cellular iron accumulation both during FAA feeding and after removal of FAA. Thus, iron exclusion by liver neoplasms is carcinogen-independent and irreversible, in contrast with that of normal hepatocytes which is completely carcinogen-dependent and reversible. Altered foci appear to represent two populations: one is characterized by reversible iron-exclusion whereas the other, like neoplasms, possesses permanent iron-exclusion. PMID- 11478315 TI - Control of DNA replication and cell proliferation in eukaryotes by aphidicolin. AB - The mycotoxin aphidicolin specifically inhibits nuclear DNA synthesis in eukaryotic cells by intereacting with the replicative DNA polymerase alpha. The drug does not bind directly to DNA nor does it interfere with RNA, protein and nucleic acid precursor synthesis. Aphidicolin offers a new tool for analyzing the mechanism of DNA replication and repair in eukaryotes and for studying the role of eukaryotic DNA polymerases. The drug might also be valuable therapeutically for controlling excessive cell proliferation without adverse effects upon non multiplying cells. We describe here how to synchronize DNA synthesis in human cells by a double block with aphidicolin and the properties of two human cell lines resistant to the drug. PMID- 11478316 TI - Nuclear structure and higher order gene structure: their role in the control of chemically-induced neoplastic transformation. AB - An overview is hereby given of the physical and physicochemical alterations at the level of nuclear structures in rat liver cells, following a chemically induced neoplastic transformation. These alterations refer to chromatin-DNA structure, from secondary (B- versus A- and Z-form) to tertiary-quanternary up to quinternary (in situ), and to the physical state of water. Possible molecular mechanisms-linking global chromatin changes to single gene expression in the control of neoplastic transformation-are discussed in terms of the degree of negative superhelicity of fibrosomes, recently identified single repeating structural subunits and hereby associated with single functional genes. PMID- 11478317 TI - Phenotyping cytochromes P450 with monoclonal antibodies. AB - Monoclonal antibodies (MAbs) to cytochrome P-450 isozymes can be used to phenotype tissues for epitope-specific cytochrome P-450 content. MAbs that inhibit specific cytochrome P-450 dependent drug or carcinogen reactions are useful tools for quantitative measurement of the individual or classes of cytochromes P-450 that catalyze these reactions. This method has been applied successfully to animal as well as human tissues. Radioimmunoassays based on MAbs have been developed and provide a rapid and efficient means for detecting cytochromes P-450 independent of functional enzyme activity. In addition, MAbs coupled to a Sepharose support can be used to immunopurify cytochromes P-450 in a procedure that is more rapid and efficient than conventional purification schemes. MAbs add a new dimension to analyses of cytochrome P-450 multiplicity and will find numerous applications in elucidation of the relationship between cytochrome P-450 phenotype and carcinogen or drug metabolism. PMID- 11478318 TI - Modulation of radiation induced transformation by combinations of a phorbol diester and a lymphotoxin. AB - The susceptibility of normal Syrian hamster embryo cells to transformation by environmental carcinogens has made possible the determination of a variety of responses as cells proceed to the neoplastic state. Expression of the initiated and promoted stages of irradiation carcinogenesis, for example, can be modified by cell surface alterations. Phytohemagglutin (PHA) or its isolectins decrease 12 O-tetradecanolphorbal-13-acetate (TPA) promoted transformation whereas PHA does not affect carcinogen only induced transformation. In contrast, both initiated and promoted transformation are sensitive to hamster lymphotoxin, a hormone-like, non-antibody lymphocyte glycoprotein. A 48 hr lymphotoxin treatment before or immediately after X-irradiation, or during TPA exposure causes a persistent inhibition independent of when carcinogen was added. A 6 hr lymphotoxin pulse before irradiation and TPA causes a persistent but nonpermanent effect unless followed by the carcinogen treatment; lymphotoxin becomes more potent as the interval between the lymphotoxin pulse and carcinogen insult or TPA addition is reduced. PHA and lymphotoxin affect the biological activity of TPA by diverse mechanisms. PHA may alter either the binding of TPA to a critical cellular receptor for promotion or alter a later step in promotion. Lymphotoxin can prevent the initiation of transformation and modulate carcinogenesis as well at both initiated and promoted stages in the transition to the neoplastic state. PMID- 11478319 TI - Orotic acid, a new promoter for experimental liver carcinogenesis. AB - Male Fischer 344 rats initiated with 1,2-dimethylhydrazine 2HCl (100 mg/kg) given 18 hr after partial hepatectomy and exposed to a diet containing 1% orotic acid for 13 months developed a 100% incidence of hepatocellular carcinoma. The creation of nucleotide pool imbalances by dietary orotic acid, for e.g., an increase in uridine nucleotides and a decrease in adenine nucleotides, was considered as a possible mechanism for the promotional effect of orotic acid on liver carcinogenesis. The significance of this hypothesis is that altered nucleotide pools affect both genomic as well as membrane organization. Consistent with this hypothesis is our finding that feeding rats with a diet containing 1% orotic acid for 10 weeks resulted in a liver DNA damage as monitored by its slower sedimentation in alkaline sucrose gradients compared to the corresponding controls. To assess the general applicability of this hypothesis, nucleotide pool imbalances were created by using methods other than feeding orotic acid and their effect on the incidence of gamma-glutamyltransferase positive foci in carcinogen initiated rats was determined. The results obtained indicated that rats initiated with 1,2-dimethylhydrazine.2HCl (100 mg/kg) given 18 hr after partial hepatectomy and exposed to diet deficient in arginine, a regimen that causes an increased synthesis and excretion of orotic acid, or were fed diets containing 1% thymidine or 1% thymine developed greater number of gamma-glutamyltransferase positive foci compared to the corresponding controls fed the basal diets. These results were interpreted to indicate that orotic acid exerts its promotional effect probably by creating an imbalance in nucleotide pools. One of the mechanisms by which an imbalance of nucleotide pools influences the pathogenesis of the carcinogenic process may be by inducing perturbations in the DNA. PMID- 11478320 TI - Fluorescence measurements of DNA-bound metabolites of benzo(a)pyrene derivatives with different carcinogenic effects. AB - (+/-)-trans-dihydroxy-7,8-dihydrobenzo(a)pyrene (BP-7,8-diol) and 9 hydroxybenzo(a)pyrene (9-OH-BP) were metabolized by rat liver microsomes in the presence of calf thymus DNA, resulting in preferential DNA binding of fluorescent (+)-anti-BP-7,8-diol-9,10-epoxide (BPDE) and 9-OH-BP-4,5-epoxide, respectively. When the DNA is denatured the fluorescence intensities of the bound metabolites change in a characteristic manner. Fluorescence decay measurements show that the intensity changes are due to changes in lifetimes of the excited states. Model substances for the bound metabolites were studied in solvents of different polarity. We found that the fluorescence changes observed after denaturation of the DNA may be explained as solvent polarity effects, so that denaturation forces the bound metabolites from a more hydrophobic environment to a hydrophilic one. Fluorescence depolarization studies as a function of temperature in combination with previous linear dichroism studies show that both BPDE and 9-OH-BP-4,5 epoxide form rigidly associated complexes with native DNA. PMID- 11478321 TI - Comparison between photo-induction and microsomal activation of polycyclic hydrocarbons with different oncogenic potency. AB - The binding of five polycyclic aromatic hydrocarbons (PAH) (anthracene (A), benzo(a)anthracene (BA), dibenz(a,h)anthracene (DBA), benzo(a)pyrene (BP) and 7,12-dimethylbenz(a)anthracene (DMBA)) to calf thymus DNA and synthetic polyribonucleotides was studied. Binding was mediated by near-ultraviolet (NUV) irradiation and 3-methylcholanthrene-induced microsomes from rat liver, in order to compare the effectiveness of these two activating systems in forming in vitro intermediates capable of binding covalently to nucleic acids. With NUV irradiation, an interaction among PAH and nucleic acids was obtained regardless of the PAH or the nucleic acid employed. The effectiveness of this activating system was higher (between 1 to 2 orders of magnitude) than that shown by induced microsomes. The enzymatic pathway bioactivated all PAH, except A, to interact with DNA. Therefore, a certain degree of correlation between the extent of DNA binding and oncogenic potency of the chemicals seemed to exist. Polynucleotide labeling was always higher than DNA labeling. PMID- 11478322 TI - Further experiments on lipid peroxidation in transplanted and experimental hepatomas. AB - The results of experiments on the subject of lipid peroxidation in hepatomas are described. It is now clear that lipid peroxidation is strongly decreased in most highly dedifferentiated hepatomas. It seems evident that the extent of the decline is strictly related to the degree of dedifferentiation. The model of diethylnitrosamine carcinogenesis, according to the method by Solt, Medline and Farber, has been now adopted to study the stages of carcinogenesis. It was shown that a net decline in lipid peroxidation occurs as early as at the stage of reversible nodules and progresses until the development of clear hepatomas. This change is practically simultaneous with a decline in the efficiency of the enzymes of the drug metabolizing system and in the content of cytochrome P450 Glutathione content and metabolism show also important changes. In fact, a dramatic increase in gamma-glutamyl-transpeptidase takes place very early during carcinogenesis, and is responsible for large decline in total glutathione during incubation of the homogenates. Glutathione peroxidase activity, on the contrary, is decreased, whereas glutathione reductase does not show significant changes. The supernatant of highly anaplastic tumors inhibits lipid peroxidation in normal liver homogenates, suggesting the presence of substances provided with antioxidant properties. These cannot be, however, related to a higher glutathione content. Supernatants from early nodules seem to be unable to block lipid peroxidation in normal liver homogenates. Preliminary experiments done to study the aldehyde pattern produced during lipid peroxidation, both in hepatomas and in nodules, confirm the presence of very poor lipid peroxidation and possibly of different peroxidation kinetics. PMID- 11478323 TI - Digital retinal photography in diabetic eye screening. PMID- 11478324 TI - Performance characteristics of digital fundus photography as a screening test for diabetic retinopathy in a low-risk population. AB - The purpose of the present study was to estimate the sensitivity and specificity of reviewers' assessments of digital fundus photography (DFP) images, using the findings from comprehensive ophthalmologic examination as the criterion standard. One hundred and fifty-two patients with diabetes underwent comprehensive ophthalmologic examination by an active duty U.S. Air Force (ADAF) staff ophthalmologist, and the examination findings were used as the criterion standard for the present study. Eight other ophthalmologists (M.D.'s) and 10 optometrists (O.D.'s), all ADAF providers, each evaluated seven nonstereo standard field DFP images from the left and right eyes (14 images per patient) of the 152 diabetic patients, assessing each patient for evidence of diabetic retinopathy (yes, no, not sure). The sensitivity, specificity, and proportion of "not sure" responses were computed separately for MD and OD reviewers and then compared. O.D. reviewers gave "not sure" responses more frequently than M.D.'s. With "not sure" responses treated as correct (i.e., identical to the results of ophthalmologic examination), both types of providers had 100% sensitivity for cases that are usually treated and 83% sensitivity for cases that are not usually treated but require close follow-up. Specificity (for any diabetic retinopathy) was 92% for M.D.'s and 93% for O.D.'s. With M.D.'s as photographic reviewers, DFP-based screening required only 27 ophthalmologic examinations per 100 patients screened; with O.D.'s as photographic reviewers, 31 ophthalmologic examinations were required per 100 patients screened. Using either ophthalmologists or optometrists as photographic reviewers, DFP-based screening for diabetic retinopathy has very good sensitivity and excellent specificity. PMID- 11478325 TI - Measurement of the glucose transport time delay between the blood and aqueous humor of the eye for the eventual development of a noninvasive glucose sensor. AB - In the recent past, several noninvasive optically based methods have been proposed for physiologic glucose sensing. One proposed optical sensing site has been the eye, which, due to its unique optical properties, can be considered as a transparent optical window into the body. In particular, the aqueous humor within the anterior chamber of the eye has been shown to contain glucose levels correlated to those of blood. Concern, however, has been expressed that using the aqueous humor solution as a measure of blood glucose may be problematic due to the potential transport time delay between the blood and the aqueous humor glucose concentrations. This investigation was performed to measure the transport time delay in a rabbit model. The time delay between the blood and aqueous humor glucose concentrations was measured invasively in five New Zealand White rabbits over a series of weeks. An anesthesia protocol containing the drug Xylazine was used to elevate the blood glucose levels to a level commonly seen in diabetic patients. The difference between the glucose peak location times occurring in the blood and aqueous humor glucose response was measured and defined as the transport time delay. The average transport time lag was measured to be under 5 min. This measured time delay indicates that, indeed, the eye could potentially be used as a sensing site for indirect blood glucose measurements and may eventually aid the development of a noninvasive glucose sensor due to its unique optical properties compared to other biological tissues. PMID- 11478326 TI - A noninvasive ocular glucose sensor. PMID- 11478327 TI - DHEA-PC slows the progression of type 2 diabetes (non-insulin-dependent diabetes mellitus) in the ZDF/Gmi-fa/fa rat. AB - The etiology of non-insulin-dependent diabetes mellitus (NIDDM) is complex and development is manifested by initial insulin resistance coupled with elevated insulin levels in the early diabetic state with concomitant increases in circulating levels of glucose and triglycerides. This is followed by a decline in insulin levels due to pancreatic exhaustion. Our results show that administration of DHEA-PC, a phosphocholine conjugate of dehydroepiandrosterone (DHEA), delayed the development of NIDDM symptoms and the onset of type 2 diabetes in the ZDF/Gmi fa/fa rat model. The treatment consisted of weekly implantation of subdermal osmotic infusion pumps in the rats starting at 6 weeks of age (n = 5 animals per group). For the first three weeks the pumps delivered 6 mg/day/rat followed by 12 mg/day/rat for 1 week (control group pumps delivered only carrier vehicle) after which the pumps were removed. Plasma was collected weekly from day 0 through day 58, and glucose, triglycerides, cholesterol, insulin, IGF-1, and IGF-BP3 levels were measured. Data were analyzed by two-way ANOVA. Following 3 weeks of treatment with DHEA-PC, plasma glucose levels in the treated group remained low, 150+/-9 mg/dL, while the levels in the control animals steadily increased to 320+/-100 mg/dL (p < 0.05). After the DHEA-PC treatment ended, plasma glucose plateaued for 10 days and then took 25 days to reach the level in the control animals (p < 0.05). After 2 weeks of DHEA-PC treatment, plasma triglyceride levels in the treated group remained low, 85+/-24 mg/dL, while the level in the control rats increased to 180+/-35 mg/dL (p < 0.05). After the treatment was terminated triglyceride levels in the treated group increased to control levels within 2 days. Insulin, IGF-1, IGF-BP3, cholesterol, body weight, and food consumption were not changed by DHEA-PC treatment (p < 0.05). Therefore, the delay of increases in plasma glucose and triglycerides, caused by DHEA-PC, was not the result of differences in caloric intake, increased insulin, or increased IGF-1 levels. The data suggest that DHEA-PC delayed the onset of the two most important parameters of NIDDM, namely hyperglycemia and hypertriglyceridemia. (ZDF/Gmi-fa/fa rats and their care was supplied by contract with Genetic Models Inc., Indianapolis, IN.). PMID- 11478328 TI - Potential therapeutic use of dehydroepiandrosterone and structural analogs. PMID- 11478329 TI - Delivery of insulin by jet injection: recent observations. AB - Medi-Ject Corporation (now Antares Pharma, Inc.) has been providing delivery devices for the needle-free administration of insulin for over 25 years. This study was one of the final steps in the development and premarket evaluation of Medi-Ject's newest needle-free system, the Medi-Jector Vision. This study was conducted to evaluate the performance of this device in the hands of experienced jet injection users in a home environment. Diabetic subjects currently using a needle-free device for the administration of their insulin were studied. Subjects used the new Medi-Jector Vision for all of their insulin administration during the course of the study. Insulin was injected on schedule and at doses consistent with their standard of practice as directed by their health care provider. All subjects were required to document each injection in a daily diary. Study subjects utilized all common insulin types (rapid, regular, intermediate, and long acting), and injections were administered in all of the common injection sites (arms, thighs, abdomen, and buttocks). Once subjects optimized the system to the most appropriate orifice size based on completeness of injection, average completeness percentages were greater than 94% for all orifice sizes. Most patients using the new Medi-Jector Vision in the home were able to manage their insulin therapy without significant complication. We conclude that the jet delivery of insulin with the new Medi-Jector Vision is well accepted by people with diabetes and offers a reliable alternative to the use of needles. PMID- 11478330 TI - Overcoming skin's barrier: the search for effective and user-friendly drug delivery. AB - Although hypodermic needles rapidly deliver large doses of drugs such as insulin across the skin for systemic administration, the pain, local trauma, and difficulty to achieve sustained or complex delivery profiles has motivated development of novel alternative technologies. Microneedles, jet injectors, and thermal poration make micron-scale holes in skin through which drugs can be driven in a user-friendly manner. Chemical enhancers, iontophoresis, electroporation, and ultrasound increase skin permeability by making submicron alterations in skin microstructure for continuous delivery over time. PMID- 11478331 TI - A novel method for continuous online glucose monitoring in humans: the comparative microdialysis technique. AB - The aim of this study was to prove the feasibility of continuous subcutaneous glucose monitoring in humans using the comparative microdialysis technique (CMT). The performance of the CMT was determined by comparing tissue glucose values with venous or capillary blood glucose values in healthy volunteers and type 1 diabetic subjects. The CMT is a microdialysis-based system for continuous online glucose monitoring in humans. This technique does not require calibration by the patient. Physiological saline with glucose (5.5 mM) is pumped in a stop-flow mode through a microdialysis probe inserted into the abdominal s.c. tissue. Tissue glucose concentration is calculated by comparing the dialysate and perfusate glucose concentrations. The time delay due to the measurement process is 9 min. We tested the CMT on six healthy volunteers and six type 1 diabetic patients for 24 h in our clinical setting. Comparisons were made to HemoCue analyzer (Angelholm, Sweden) capillary blood glucose measurements (healthy volunteers) and to venous blood glucose concentration determined with a Hitachi analyzer (diabetic patients). The mean absolute relative error of the CMT glucose values from the blood glucose values was 17.8+/-15.5% (n = 167) for the healthy volunteers and 11.0+/-10.8% (n = 425) for the diabetic patients. The mean difference was 0.42+/-1.06 mM (healthy volunteers) and -0.17+/-1.22 mM (diabetic patients). Error grid analysis for the values obtained in diabetic patients demonstrated that 99% of CMT glucose values were within clinically acceptable regions (regions A and B of the Clarke Error Grid). The study results show that the CMT is an accurate technique for continuous online glucose monitoring. PMID- 11478332 TI - Towards retrievable vascularized bioartificial pancreas: induction and long lasting stability of polymeric mesh implant vascularized with the help of acidic and basic fibroblast growth factors and hydrogel coating. AB - We seek to improve existing methodologies for allogenic grafting of pancreatic islets. The lack of success of encapsulated transplanted islets inside the peritoneal cavity is presently attributed to poor vascularization of the implant. A thick, fibrotic capsule often surrounds the graft, limiting survival. We have tested the hypothesis that neovascularization of the graft material can be induced by the addition of proper angiogenic factors embedded within a polymeric coat. Biocompatible and nonresorbable meshes coated with hydrophilic polymers were implanted in rats and harvested after 1-, 6-, and 12-week intervals. The implant response was assessed by histological observations on the degree of vascularity, fibrosis, and inflammation. Macrostructural geometry of meshes was conducive to tissue ingrowth into the interstitial space between the mesh filaments. Hydrogel coating with incorporated acidic or basic FGF in an electrostatic complex with polyelectrolytes and/or with heparin provided a sustained slow release of the angiogenic growth factor. Anti-factor VIII and anti collagen type IV antibodies and a GSL I-B4 lectin were used to measure the extent of vascularization. Vigorous and persistent vascularization radiated several hundred microns from the implant. The level of vascularization should provide a sufficient diffusion of nutrients and oxygen to implanted islets. Based on our observations, stable vascularization may require a sustained angiogenic signal to allow for the development of a permanent implant structure. PMID- 11478333 TI - The relation of type 2 diabetes and cancer. AB - Increasing evidence indicates that individuals with type 2 diabetes (diabetes) are at elevated risk for several common human malignancies, including cancers of the colon, breast, endometrium, pancreas, and liver. In particular, the consistent positive results reported by prospective investigations make it unlikely that methodologic issues, occult tumors, or chance results could explain the findings. Since diabetes and impaired fasting glucose together affect >25% of Americans above age 50, even a moderate etiologic association (e.g., relative risk = 1.5) would explain >10% of involved malignancies. Laboratory studies have suggested biologically plausible mechanisms. Insulin, for example, is typically at high levels during the development and early stages of diabetes. Activation of the insulin receptor by its ligand, or cross-activation of the insulin-like growth factor-I receptor, has been shown to be mitogenic and promote tumorigenesis in various model systems. A "unifying concept," in fact, holds that hyperinsulinemia may underlie the cancer associations of several additional risk factors, including high waist circumference, visceral fat, waist-to-hip ratio, body mass index, sedentary lifestyle, and energy intake. In this review, we assess current evidence regarding the relation of type 2 diabetes with cancer, and evaluate the findings in terms of well-accepted criteria for establishing causality. PMID- 11478334 TI - FDA approves new glucose test for adult diabetics. PMID- 11478335 TI - Special funding boosts type 1 diabetes research. PMID- 11478336 TI - Watching your glucose with the GlucoWatch. PMID- 11478337 TI - The HIPAA privacy rule: its effect on diabetes treatment. Health Insurance Portability and Accountability Act. PMID- 11478338 TI - Medicare ruling is a devastating setback for diabetes education. PMID- 11478339 TI - Questionnaires for a randomized controlled trial methodology to evaluate the teaching utility of diabetes simulation programs. AB - AIDA is an interactive educational diabetes simulator which has been made available without charge on the Internet. Since its launch on the World Wide Web in 1996 over 87,000 people have visited the AIDA Website--http://www.2aida.org- and over 22,000 copies of the program have been downloaded from there free-of charge. The AIDA software is believed to be of use in recreating clinically realistic diabetes situations for interactive simulation. However, despite its widespread application, its actual utility for supporting the education of patients with type 1 diabetes mellitus remains to be objectively demonstrated in a clinical randomised controlled trial (RCT) setting. The current "Diabetes Information Technology & WebWatch" column overviews a prospective RCT methodology for formally evaluating the educational utility of an interactive diabetes simulator, like AIDA. The protocol makes use of two study arms--each receiving different educational interventions. During lessons, arm A of the study will be exposed to the AIDA simulator (the active intervention), while arm B (the control group) will benefit from conventional educational methods using standard presentations with slides and transparencies. Six lessons will be held for each study arm (one per week). At the beginning and end of the study self-monitoring blood glucose (SMBG) data will be collected, details of any hypoglycemic episodes recorded, and assessments made of glycosylated hemoglobin (HbA1c) levels. Participants will also be required to complete a detailed questionnaire to assess their self-confidence, quality of life and metabolic control, attitudes towards SMBG, and knowledge about insulin dosage calculation. Comparisons will be made between arm A and arm B using unpaired statistical analyses. A partial cross-over study design is also proposed whereby subsequently the control group will be exposed to the AIDA simulator during a further 6-week course of lessons. This will ensure that the maximum number of subjects will eventually receive the active intervention, and will also allow further within group paired analyses to be applied (with greater statistical power). This current "Diabetes Information Technology & WebWatch" column documents two of the questionnaires which are intended to be used for this RCT approach. PMID- 11478340 TI - Evaluating health-care delivery: Hospital in the Home. PMID- 11478341 TI - Individualized treatment of heart failure. PMID- 11478342 TI - Differences in length of stay for Hospital in the Home patients: comparing simple clinical coding with medical record review. AB - AIM: To determine whether the apparently longer length of stay (LOS) reported for patients with cellulitis managed in Hospital in the Home (HITH) compared with those managed as inpatients was correct. METHODS: Data, including LOS, from the Victorian In-patient Minimum Database (VIMD) of all patients with cellulitis managed between July 1998 and June 1999 at a large metropolitan teaching hospital were analysed and compared with a retrospective medical record review of the same patients. RESULTS: In the VIMD data, there were 266 episodes of cellulitis during the study period. However, the medical record review found that six episodes were not separate, but rather a continuation of treatment for the same episode of cellulitis, and that 18 were not episodes of cellulitis, but were pilonidal sinus infections. In the VIMD data set, the mean LOS for patients treated in HITH was generally longer than that for inpatients (7.2 days vs 5.1 days, respectively, P = 0.002). However, in the retrospective medical record review, the LOS for patients treated in HITH was similar to inpatients (7.3 days versus 7.0 days, respectively, P = 0.68). CONCLUSIONS: In contrast to the VIMD data, the medical record review demonstrated that, overall, patients with cellulitis had a similar LOS irrespective of whether they were managed at home or in hospital. This study confirms that caution is required in interpreting the VIMD data, highlights the importance of carefully monitoring the introduction of new treatment modalities and indicates areas for further research. PMID- 11478343 TI - Early Australian experience with infliximab, a chimeric antibody against tumour necrosis factor-alpha, in the treatment of Crohn's disease: is its efficacy augmented by steroid-sparing immunosuppressive therapy? The Infliximab User Group. AB - BACKGROUND: Tumour necrosis factor-alpha (TNF-alpha) plays an important role in the pathology of Crohn's disease. Infliximab, a chimeric antibody against TNF alpha, has been shown in controlled clinical trials to be effective in two-thirds of patients with refractory or fistulating Crohn's disease. The factors that determine a clinical response in some patients but not others are unknown. AIMS: To document the early Australian experience with infliximab treatment for Crohn's disease and to identify factors that may determine a beneficial clinical response. METHODS: Gastroenterologists known to have used infliximab for Crohn's disease according to a compassionate use protocol were asked to complete a spreadsheet that included demographic information, Crohn's disease site, severity, other medical or surgical treatments and a global clinical assessment of Crohn's disease outcome, judged by participating physicians as complete and sustained (remission for the duration of the study), complete but unsustained (remission at 4 weeks but not for the whole study) or partial clinical improvement (sustained or unsustained). RESULTS: Fifty-seven patients were able to be evaluated, with a median follow-up time of 16.4 (4-70) weeks, including 23 patients with fistulae. There were 21 adverse events, including four serious events. Fifty-one patients (89%) had a positive clinical response for a median duration (range) of 11 (2-70) weeks. Thirty patients (52%) had a remission at 4 weeks, 10 of whom had remission for longer than 12 weeks. Forty-two per cent of fistulae closed. Sustained remission (P = 0.065), remission at 4 weeks (P = 0.033) and a positive clinical response of any sort (P = 0.004) were more likely in patients on immunosuppressive therapy, despite there being more smokers in this group. CONCLUSION: This review of the first Australian experience with infliximab corroborates the reported speed and efficacy of this treatment for Crohn's disease. The excellent response appears enhanced by the concomitant use of conventional steroid-sparing immunosuppressive therapy. PMID- 11478344 TI - Urinary symptoms and incontinence in an urban community: prevalence and associated factors in older men and women. AB - BACKGROUND: There is increasing recognition of the importance of a wide range of urinary symptoms in both men and women and that these symptoms are undertreated. AIMS: To determine the prevalence of and factors associated with urinary symptoms, including nocturia, urgency, urge and stress incontinence and, in men, urinary stream difficulties; and the prevalence of being bothered by the symptoms and ever seeking treatment for them. METHOD: Household survey by computer assisted telephone interviews of people aged 41 years and over and living in inner metropolitan Sydney. RESULTS: Fifty-three per cent (95% confidence interval (CI) 46-60) of men and 61% (95% CI 55-67) of women reported one or more symptoms in the previous month. In men, the most frequently reported symptoms were urgency (30%, 95% CI 24-36) and nocturia (25%, 95% CI 19-31). In women, stress incontinence (35%, 95% CI 29-41) and urgency (33%, 95% CI 27-39) were the most common symptoms reported. In men, the significant factors associated with reporting one or more symptoms, after adjustment for other variables, were age 60 years or more, no private medical insurance, obesity and fair or poor self-rated health. For women, the significant associations were age 50-59 years, age 70 years or more, no private health insurance, high psychological distress and fair or poor self-rated health. CONCLUSIONS: Urinary symptoms are experienced by more than half of men and women aged over 40 in the central Sydney community, but many do not seek treatment. Such symptoms should be considered more broadly than the traditional focus on male 'prostatism' and female incontinence. PMID- 11478345 TI - Prevalence survey of multiple sclerosis in the Australian Capital Territory. AB - AIM: This study sought to obtain an estimate of the prevalence of multiple sclerosis (MS) in the Australian Capital Territory (ACT), a largely urban region that differs climatically and socioeconomically from other Australian cities examined in previous MS surveys. METHODS: Prevalence day was chosen to coincide with the 1996 National Census. All ACT neurologists' records for the previous 5 years were examined and cases of MS were classified according to the published diagnostic criteria of Rose et al. and Poser et al. RESULTS: By the criteria of Rose et al., as used in previous Australian surveys of MS, prevalence was 79.9/100,000 (95% confidence interval (CI) = 63.4-99.2) for females, 32.8 (22.7 46.2) for males and 56.7 (43.1-74.1) for all people, standardized to the 1996 population. Standardized to the 1981 population for direct comparison with 1981 surveys in New South Wales, the prevalence of MS in the ACT was still unexpectedly high, particularly for females. Using the criteria of Poser et al., the prevalence of MS standardized to the 1996 population was 70.6/ 100,000 (95% CI = 58.4-85.3) for females, 28.0 (20.3-37.8) for males and 49.5 (42.2-58.2) for all people. There was evidence from a relatively short duration of disease in the ACT sample that some persons with long-standing MS had been missed in the survey and therefore that the prevalence of MS observed in the ACT was an underestimate. CONCLUSIONS: The survey found an unexpectedly high prevalence of MS in the ACT. Possible reasons for this are discussed. There was no evidence that the advent of magnetic resonance imaging had increased the numbers of persons diagnosed with MS in the present survey. PMID- 11478346 TI - Early rheumatoid arthritis: can we predict its outcome? AB - The continuing trend towards more aggressive treatment of rheumatoid arthritis (RA) has seen an increasing interest in the early phase of this chronic inflammatory disease. Optimal benefit from present and emerging therapies is limited by our prognostic abilities during this period. The present review attempts to outline first the many methodological issues encountered in studies of early RA, and second the extent to which each major outcome measure can be explained, both by readily available clinical variables and by HLA-DR genotyping. The evidence supporting the clinical usefulness of genotyping is discussed separately. Based on this information, a clinically appropriate approach to the management of early RA and the identification of patients suitable for experimental therapies is suggested. PMID- 11478347 TI - Is there another chapter in the Helicobacter pylori/peptic ulcer disease story? AB - The role of Helicobacter pylori in the production of mucosal damage has largely been considered within a simple infection paradigm, because to date eradication has appeared to be a predictable outcome of antibiotic therapy. Changes in the epidemiology and management of peptic ulcer disease, however, require a more comprehensive framework to understand these shifting clinical patterns. The present review examines mucosal damage as an outcome of a complicated host parasite relationship, with alterations in both parasite physiology and host defence mechanisms being keys to understanding disease patterns. PMID- 11478348 TI - On patient judgement. AB - Clinical judgement, the keystone of medical expertise, is a hot topic. By contrast, patient judgement, also of central importance in health care, receives little attention. Patients have the last say concerning whether or not they seek medical treatment for symptoms, follow a doctor's advice or accept reassurance. Delay in seeking help for serious symptoms, non-compliance with treatment advice and failure of doctors to reassure many of the 'worried well' have long been recognized as serious problems. We argue that what is common to these important problems is patient judgement. Surveys yielding information about the average influence of a large number of individual variables do not do justice to the complex interaction of influences that can influence the judgements of an individual person under particular social circumstances. This is what explains the wide variety of patient reactions. From the medical standpoint, such unpredictable patient behaviour seems irrational. The patient perspective on the meaning of their actions is a hiatus in our knowledge, which is hampering the planning of effective interventions. Too few studies have sought the perspective of patients by asking them why they acted as they did. Thus, the wide spectrum of patient response in these situations in relation to personality, life experience and social context cannot be studied without interpretive field studies that include interview of patients with qualitative interpretation. PMID- 11478349 TI - History of virology in Australia: selected highlights. PMID- 11478350 TI - Recurrent ischaemic colitis associated with pseudoephedrine use. PMID- 11478351 TI - Chemotherapy and granulocyte colony stimulating factor-mobilized blood cell infusion followed by interferon-alpha for relapsed malignancy after allogeneic bone marrow transplantation. AB - BACKGROUND: Interferon-alpha (IFN) is known to promote graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo BMT). This property may also be used to enhance a graft-versus-leukaemia effect (GVL) after donor leucocyte infusion (DLI), a mode of therapy increasingly offered to patients relapsing after allo BMT. AIM: The aims of the present study were to examine the efficacy and toxicity of IFN therapy administered after granulocyte colony stimulating factor (G-CSF)-stimulated blood cells given as DLI in patients with acute myeloid leukaemia (AML), chronic myeloid leukaemia (CML), acute lymphoblastic leukaemia (ALL), acute undifferentiated leukaemia (AUL) and multiple myeloma relapsing after allo BMT. METHODS: Between October 1996 and September 1999, 27 patients (16 AML, four ALL, three CML, three multiple myeloma, one AUL) who relapsed after allo BMT were treated with chemotherapy followed by DLI, collected after G-CSF stimulation in all but two cases. Subsequently, IFN was given to patients without significant GVHD or rapidly progressive disease. The outcome after DLI with regard to remission rate, disease-free survival and GVHD was analysed. RESULTS: Eighteen patients received IFN following DLI, 14 of whom developed significant GVHD (grade II-IV acute or extensive chronic); thereafter, GVHD resolved with cessation of IFN alone in four patients, but 10 required systemic immunosuppression. Twenty-three patients were given chemotherapy and DLI as initial treatment of relapse; 10 achieved complete remission (CR), in four patients this was only after the onset of GVHD. The other four patients received chemotherapy and DLI as a consolidation of a chemotherapy induced remission. The CR was durable only in patients with CML (3 of 3) and AML (4 of 8). CONCLUSIONS: Treatment with IFN induced GVHD in the majority of patients receiving DLI. The induction of GVHD and GVL by this approach produced excellent results in patients with CML and modest results in AML, but appeared to be less effective in myeloma and ALL. PMID- 11478352 TI - Aminoglycosides in cystic fibrosis: a descriptive study of current practice in Australia. AB - AIM: To determine the diversity of clinical practice with respect to aminoglycosides in cystic fibrosis (CF) units within Australia. METHOD: In April 1999, a questionnaire on the use of aminoglycosides was sent to 30 CF units across Australia. Information was collected about drug selection, dosing, monitoring and toxicity with intravenous administration. RESULTS: Completed surveys were received from 26 of the 30 units (response rate = 86%) and all units with > 40 patients. Tobramycin was the drug of choice in all but two centres where there was equivalent use of gentamicin and tobramycin. The survey demonstrated a trend in recent years to reduce the number of doses per day with 54% of centres prescribing once daily, 23% twice daily and 23% thrice daily regimens. Initial dosing was generally based on mg/kg per day (mean 8.8, range 7.5-10 mg/kg per day). Dosing by infusion occurred in 11 of 14 units using once daily dosing and there was equivalent use of bolus and infusion methods for multiple-daily regimens. Drug monitoring depended on dosing regimen. Units using multiple daily regimens monitored using trough +/- peak levels, whereas 50% of units using once-daily dosing used two postdose levels to alter dose. Actual toxicity, in particular nephrotoxicity, ototoxicity and vestibular toxicity was reported by 19, 27 and 12% of units, respectively. CONCLUSION: The prescribing, dosing and monitoring of aminoglycosides in CF across Australia varies greatly. This is likely to be due to a lack of definitive evidence as to the optimum use in this patient group. PMID- 11478353 TI - Current status of positron emission tomography in oncology. AB - Positron emission tomography (PET) has emerged as a powerful diagnostic tool in oncology patients. There is evidence of the superior utility over conventional imaging methods of the principal PET tracer 18F-fluoro-2-deoxy-glucose in the staging of a range of cancers and monitoring disease recurrence, as well as changing patient management to more appropriate therapy. The methods for evaluating the evidence for PET remain complex, particularly as the standard evidence-based approach of randomized controlled trials is not generally applicable to imaging technologies. PET has the potential to dramatically improve our ability to manage patients with cancer and is also making major contributions to the development of new therapies. PMID- 11478354 TI - Fluoro-2-deoxy-glucose positron emission tomography in oncology: are we there yet? PMID- 11478355 TI - Cyclooxygenase inhibitors: any reservations? AB - The discovery of two cyclooxygenase isozymes, constitutive COX-1 and inducible COX-2, has resulted in the rapid development of selective inhibitors of COX-2, such as celecoxib and rofecoxib. Compared with traditional non-steroidal anti inflammatory drug agents, use of COX-2 selective inhibitors is associated with decreased incidence of adverse gastric events as a result of minimal inhibition of gastroprotective COX-1, but with equivalent anti-inflammatory benefit through inhibition of COX-2. However, there is evidence to suggest that the 'COX-1 = constitutive, COX-2 = inflammatory' paradigm is less distinct than originally proposed. Futhermore, selective COX-2 inhibitors may have other consequences as a result of the change in the eicosanoid profile. Thus, despite the relatively safe gastrointestinal profile, vigilant post-marketing surveillance for other adverse effects is required. PMID- 11478356 TI - Patent foramen ovale and cryptogenic cerebral infarction. AB - The patent foramen ovale (PFO) has been increasingly implicated in the aetiology of stroke, particularly in young patients with no other identifiable cause (cryptogenic stroke). The mechanism is by the passage of venous clots through the patent foramen into the arterial circulation, enabling cerebral embolism. Such patients with cryptogenic stroke and PFO are often treated with life-long anticoagulants or antiplatelet agents in an attempt to decrease the risk of a recurrence. Less commonly, surgical closure of the PFO has been undertaken in these patients. However, the recent evolution of closure devices that are delivered percutaneously by standard cardiac catheter techniques now offer an alternative non-surgical option. These alternative therapies are yet to be compared adequately. Two issues remain to be resolved. First, in determining appropriate therapy, patients with cryptogenic stroke may be divided into three groups: those with no PFO but an alternative unrecognized aetiology, those with an 'innocent' PFO and an alternative unrecognized aetiology and those with a causative PFO. The distinction between these groups clearly has important treatment implications. Second, the risk versus benefit of each available treatment modality must be determined for these different patient subgroups. These two issues require resolution before rational evidence-based treatment can be prescribed for patients with PFO and cryptogenic stroke. PMID- 11478357 TI - Population screening for HFE-associated haemochromatosis: should we have to pay for our genes? AB - Haemochromatosis associated with mutations in the HFE gene is the most common inherited disorder in Caucasian populations. Early diagnosis and treatment allows for normal life expectancy, whereas there is considerable morbidity and early mortality in those patients diagnosed late or untreated. Unfortunately, the development of symptoms and signs in haemochromatosis is usually associated with significant iron overload. For this reason, many clinicians and geneticists have advocated population screening. The recent identification of the HFE gene and the availability of a simple DNA-based diagnostic test have led to international debate as to the most cost-effective means of population screening for HFE associated haemochromatosis. The present paper summarizes the evidence in favour of population screening and analyses the relative advantages of genotypic (DNA test) versus phenotypic (transferrin saturation) testing. PMID- 11478358 TI - Enterococci resistant to vancomycin. PMID- 11478359 TI - Molecular medicine in the 21st century. AB - When Watson and Crick proposed the double helix model for DNA structure in a 2 page Nature article in 1953, no one could have predicted the enormous impact this finding would have on the study of human disease. Over the last decade in particular, major advances have been made in our understanding of both normal biological processes and basic molecular mechanisms underlying a variety of medical diseases. Knowledge obtained from basic cellular, molecular and genetic studies has enabled the development of strategies for the modification, prevention and potential cure of human diseases. This brief overview focuses on the enormous impact molecular studies have had on various aspects of medicine. The inherited cardiac disorder hypertrophic cardiomyopathy is used here as a model to illustrate how molecular studies have not only redefined 'gold standards' for diagnosis, but have also influenced management approaches, increased our understanding of fundamental disease-causing mechanisms and identified potential targets for therapeutic intervention. The near-completion of the Human Genome Project, which identifies the 3.2 billion base pairs that comprise the human genome (the so-called 'Book of Life'), has exponentially heightened the focus on the importance of molecular studies and how such studies will impact on various aspects of medicine in the 21st century. PMID- 11478360 TI - Does reading poetry make you a better clinician? PMID- 11478361 TI - Stuttering myocardial infarction caused by spontaneous coronary dissection. PMID- 11478362 TI - Scleroderma cardiomyopathy presenting with thromboembolism. PMID- 11478363 TI - Combined heart and liver transplantation for familial amyloidosis. PMID- 11478364 TI - Quadriceps muscle wasting persists 5 months after total hip arthroplasty for osteoarthritis of the hip: a pilot study. AB - AIMS: To determine whether additional muscle fibre wasting of the ipsilateral vastus lateralis muscle occurs in the early postoperative period after total hip arthroplasty for osteoarthritis of the hip and whether there is an improvement in preoperative measures of quadriceps muscle thickness, strength, pain and function over a 5-month postoperative period. METHODS: Twelve patients had ipsilateral needle quadriceps biopsy for muscle morphology and bilateral quadriceps muscle thickness ultrasound preoperatively, 5 days and 4 weeks postoperatively and a further muscle thickness measurement at 5 months. Seven additional patients and five age-matched control subjects had bilateral quadriceps muscle ultrasound thickness preoperatively, 6 weeks and 5 months postoperatively, with assessment of quadriceps muscle dynamometry, pain scores and Timed Up and Go (TUG) test. RESULTS: Preoperatively, all 19 patients demonstrated significant atrophy of the ipsilateral compared with the contralateral quadriceps muscle (P = 1.8 x 10(-7)) on muscle ultrasound, which persisted at 5 months follow up (P = 0.009). Muscle morphology preoperatively showed type 2A and 2B muscle fibre atrophy on needle muscle biopsy, with further atrophy of all three fibre types (P = 0.029) at 5 days postoperatively associated with a fibre type shift from type 1 to 2A fibres (P = 0.0011) at 1 month. There was improvement in hip pain postoperatively and a significant improvement in the TUG test (P = 0.007). However, there was no improvement in muscle strength on dynamometry. CONCLUSIONS: There is significant ipsilateral quadriceps atrophy and weakness with 2A and 2B fibre atrophy preoperatively in patients with osteoarthritis of the hip with exacerbation and further atrophy of all three fibre types 5 days postoperatively. Postoperative follow up showed that the reduction in ipsilateral quadriceps muscle thickness persisted at 5 months despite physical rehabilitation. Patients did note significant improvement in pain postoperatively and improvement on functional assessment with the TUG test. Other therapeutic strategies may have to be developed to reverse disuse muscle atrophy. PMID- 11478365 TI - Cellular transport of nonesterified fatty acids. AB - Transport of nonesterified fatty acids (NEFA) is an important component of whole body energy metabolism, and derangements in NEFA transport have been linked to several diseases. NEFA are transferred from their sites of production to cells in hepatic and peripheral tissues by mechanisms that are regulated in part by cell status and as determined by the covalent structure of the NEFA species. Major barriers to physical transport are transfer from the hydrophobic surfaces on cell membranes and NEFA-binding proteins, such as albumin, into the surrounding aqueous phase and translocation across a membrane that contains a very hydrophobic interior; this process could be purely diffusive or require specific protein cofactors. Herein evidence is provided suggesting that this step is driven by intracellular metabolism that supports a NEFA gradient across the cell membrane. According to current models of NEFA transfer, the rate-limiting step is likely to be desorption of NEFA from the inner leaflet of the cell membrane or intracellular metabolism; for very long chain NEFA, the former is more likely. PMID- 11478366 TI - Role of CD36 in membrane transport and utilization of long-chain fatty acids by different tissues. AB - The transmembrane glycoprotein CD36 has been identified in isolated cell studies as a putative transporter of long-chain fatty acids. To examine the physiological role of CD36, we studied FA uptake and metabolism by tissues of CD36 null mice after injection with two fatty acid analogs. Compared to controls, uptake was substantially reduced (50-80%) in heart, skeletal muscle, and adipose tissues of null mice. The reduction in uptake was associated with a large decrease in fatty acid incorporation into triglycerides, which could be accounted for by an accumulation of diacylglycerides. Thus CD36 facilitates a major fraction of fatty acid uptake by myocardial, skeletal muscle, and adipose tissues, where it is highly expressed. Its role in other tissues where its expression is low and cell specific could not be determined in these studies. PMID- 11478367 TI - Involvement of membrane-associated proteins in the acute regulation of cellular fatty acid uptake. AB - The transport of long-chain fatty acids across cellular membranes most likely occurs to some extent by passive diffusion and additionally is facilitated by a number of membrane-associated and cytoplasmic proteins. In this overview we focus on the involvement of the membrane proteins fatty acid translocase (FAT/CD36), plasma membrane fatty acid-binding protein (FABPpm) and fatty acid-transport protein (FATP). Newly obtained evidence is presented that in skeletal muscle, fatty acid uptake is subject to short-term regulation by translocation of FAT/CD36 from intracellular stores to the plasma membrane, analogous to the regulation of muscular glucose uptake by GLUT-4 translocation. These new findings establish a significant role of membrane-associated proteins in the cellular fatty acid-uptake process. Possible implications for the uptake and transport of long-chain fatty acids by the brain are discussed. PMID- 11478368 TI - Fatty acid-binding proteins of nervous tissue. AB - Fatty acid-binding proteins (FABPs) are cytosolic 14-15 kDa proteins, which are supposed to be involved in fatty acid (FA) uptake, transport, and targeting. They may modulate FA concentration and in this way influence function of enzymes, membranes, ion channels and receptors, and gene expression and cellular growth and differentiation. Nine FABP types can be discerned with a specific tissue distribution. In spite of 30-70% amino acid sequence identity, they have a similar tertiary, beta-clam structure in which the FA is bound. Nervous tissue contains four FABP types with a distinct spatio-temporal distribution. Myelin (M) FABP is only present in the peripheral nerves, brain (B)-FABP and epidermal (E) FABP mainly in glial cells and neurons, respectively of pre- and perinatal brain, and heart (H)-FABP in adult brain. Possible functions of FABPs in the nervous system are discussed. Binding studies with a range of physiological FA showed no large differences between recombinant proteins of the four human FABP types in binding specificity and affinity, also not for polyunsaturated FA (PUFA). The transfer of FA from fixed liposomes to mitochondria was similarly promoted by the four types. A marked difference in conformational stability was observed with H FABP > B-FABP > M-FABP > E-FABP. Surface epitopes of H-FABP showed reaction with anti-B-FABP antibodies, but no other cross-reactivity of FABP type and heterologous antibodies was observed. The functional significance of the distinct spatio-temporal pattern of the four FABP types remains to be elucidated. PMID- 11478369 TI - Collision-mediated transfer of long-chain fatty acids by neural tissue fatty acid binding proteins (FABP): studies with fluorescent analogs. AB - Mammalian fatty acid-binding proteins (FABP) are a family of intracellular proteins (approx 15 kDa) that bind long-chain fatty acids (FA) with high affinity. They are believed to serve as cytoplasmic transporters of FA and to target FA to specific cellular sites of utilization. Several different FABPs are expressed in neural tissue, including brain FABP (B-FABP), myelin FABP (M-FABP), and heart FABP (H-FABP). We have previously shown that H-FABP transfers FAvia direct collisional interactions with acceptor model membranes. In the present studies, we use a fluorescence resonance energy transfer (FRET) assay to examine the rate and mechanism of transfer of a fluorescent long-chain fatty acid from B FABP to phospholipid vesicles. The rate of transfer is shown to be independent of buffer ionic strength and dramatically enhanced by the presence of specific anionic phospholipids. These results are consistent with a mechanism by which FA are transferred from B-FABP to phospholipid membranes by a transient collision based mechanism. PMID- 11478370 TI - Plasma free fatty acid and lipoproteins as sources of polyunsaturated fatty acid for the brain. AB - Polyunsaturated fatty acids (PUFA), which comprise 25-30% of the fatty acids in the human brain, are necessary for normal brain development and function. PUFA cannot be synthesized de novo and must be supplied to the brain by the plasma. It is necessary to know the PUFA content and composition of the various plasma lipids and lipoproteins in order to understand how these fatty acids are taken up and metabolized by the brain. Human plasma free fatty acid (FFA) ordinarily contains about 15% linoleic acid (18:2n-6) and 1% arachidonic acid (AA) (20:4n 6). Plasma triglycerides, phospholipids, and cholesterol esters also are rich in linoleic acid, and the phospholipids and cholesterol esters contain about 10% AA. These findings suggest that the brain probably can obtain an adequate supply of n 6 PUFA from either the plasma FFA or lipoproteins. By contrast, the plasma ordinarily contains only one-tenth as much n-3 PUFA, and the amounts range from 1% alpha-linolenic acid (18:3n-3) in the plasma FFA to 2% docosahexaenoic acid (22:6n-3, DHA) in the plasma phospholipids. The main n-3 PUFA in the brain is DHA. Therefore, if the plasma FFA is the primary source of fatty acid for the brain, much of the DHA must be synthesized in the brain from n-3 PUFA precursors. Alternatively, if the brain requires large amounts of preformed DHA, the phospholipids contained in plasma lipoproteins are the most likely source. PMID- 11478371 TI - Fatty acid uptake and incorporation in brain: studies with the perfusion model. AB - The contributions of individual components of blood to brain [14C]palmitate uptake and incorporation were studied with the in situ brain perfusion technique in the pentobarbital-anesthetized rat. With whole-blood perfusate, brain unacylated [14C]palmitate uptake was linear with time and extrapolated to zero at T = 0 s of perfusion. Tracer accumulated in brain with a blood-to-brain transfer coefficient of 1.8 +/- 0.1 x 10(-4) mL/s/g (whole cerebral hemisphere). Incorporation into brain lipids was rapid such that approximately 40% of tracer in brain at 45 s of perfusion was in cerebral phospholipids and neutral lipids. Similar rates of uptake were obtained during unacylated [14C]palmitate perfusion in whole rat plasma, serum, or artificial saline containing 2-3% albumin, suggesting that albumin has a key role in determining [14C]palmitate uptake in brain. The excellent match in brain uptake rates between whole blood and albumin containing saline fluid suggests that the perfusion technique will be useful method for quantifying the individual contributions of blood constituents and albumin binding on brain [14C]palmitate uptake. PMID- 11478372 TI - NMR and isotope ratio mass spectrometry studies of in vivo uptake and metabolism of polyunsaturates by the developing rat brain. AB - This article describes the application of in vivo 13C-nuclear magnetic resonance (NMR) spectroscopy and gas chromatography (GC)-combustion-isotope ratio mass spectrometry to the study of brain uptake and metabolism of polyunsaturated fatty acids in the suckling rat model. NMR spectroscopy is uniquely suited to the non invasive detection of nonradioactive metabolites in living animals. We applied this approach to the noninvasive detection of 13C-arachidonate in brain and liver of living suckling rats but found that technical limitations in our model, mainly poor signal-to-noise, largely prevent useful results at this time. However, in a tracer study using simultaneous doses of 13C-gamma-linolenate and 13C arachidonate, 13C-NMR of tissue lipid extracts quantitatively demonstrated a 10 fold greater (liver) or 17-fold greater (brain) accumulation of pre-formed vs newly synthesized arachidonate. GC-combustion-isotope ratio mass spectrometry was used to trace the utilization of [U-13C]-alpha-linolenate into three products in the brain: docosahexaenoate, cholesterol, and palmitate. The rationale was that although alpha-linolenate is used in de novo lipogenesis, the quantitative importance of this pathway is unknown. Our results in the suckling rat show that 2-13% of carbon from [U-13C]-alpha-linolenate appearing in brain lipids is in docosahexaenoate while the rest is in brain lipids synthesized de novo. Overall, these results indicate that the suckling rat brain prefers pre-formed to newly synthesized arachidonate and that alpha-linolenate is readily utilized in brain lipid synthesis. These methods are suited to comparative studies of the metabolism of polyunsaturates and they support previous observations that the metabolism of some polyunsaturates such as alpha-linolenate extends well beyond the traditional desaturation-chain elongation pathway. PMID- 11478373 TI - Essential polyunsaturated fatty acids and the barrier to the brain: the components of a model for transport. AB - Several areas of research have contributed to the establishment of a paradigm that meets the requirements for the selective uptake of essential polyunsaturated fatty acids (EPUFA) into brain. First, discrete studies have demonstrated that cholesterol and the nonessential fatty acids, (palmitic, oleic, stearic) do not enter the brain parenchyma. These studies demonstrated that the 18 carbon monocarboxylic fatty acids, linoleic acid with two cis-double bonds entered brain, whereas oleic acid, with one cis-double bond, did not enter brain. It was concluded the entry of essential fatty acids into brain is accomplished in a highly selective and discrete manner. Further, the typical blood-borne lipoproteins do not traverse the endothelial cells of the capillary network and enter into the brain, otherwise cholesterol, palmitic, oleic, and stearic acids from blood would be located within brain. Second, several investigators have shown that the endothelial cells of the capillary network contain lipoprotein receptors, yet one conclusion is that the brain does not utilize low-density lipoprotein (LDL)-cholesterol. Third, recently, the existence and function of a significant number of distinctive trans-membrane monocarboxylic acid transporters, (MCTs) and fatty acid transport proteins (FATPs) have been described. No transporters have been described to date with the specificity necessary to transfer only EPUFA into brain. A blueprint with the minimal elements for delivery and selectivity is proposed. Lipoproteins enter the endothelial cells because the lipoprotein receptors are positioned on their luminal membrane. Essential fatty acid transporter(s) are positioned on the abluminal membrane of these endothelial cells to allow for the entry of EPUFA into brain. Within the endothelial cell there is opportunity for lipid management and transformation such that EPUFAs are selectively culled for delivery to the essential fatty acid transporter(s), which facilitates their transfer into brain. PMID- 11478374 TI - Polyunsaturated fatty acid synthesis and release by brain-derived cells in vitro. AB - The brain is more highly enriched than most other tissues in long-chain polyunsaturated fatty acids (PUFA), particularly docosahexaenoic acid (DHA). In vitro studies of PUFA synthesis and release utilizing cell cultures of astrocytes, neurons, and cerebral microvascular endothelium have contributed significantly to our understanding of mechanisms potentially involved in the accretion of PUFA in brain. Both cerebral endothelium and astrocytes avidly elongate and desaturate precursors of the long-chain PUFAs when grown individually or in various co-culture combinations. The products, such as arachidonic acid (AA) and DHA, are released from the cells. In contrast, neurons appear unable to carry out fatty acid desaturation and thus are dependent upon preformed long-chain PUFA. Indeed, neurons co-cultured with astrocytes accumlate docosahexaenoate synthesized by the glial cells. Cerebral endothelial cultures are additionally capable of enriching the basolateral compartment (analogous to the brain extracellular space) with n-3 PUFA when grown in a membrane/chamber apparatus. The enrichment of this compartment with DHA is increased when cerebral endothelium is co-cultured with astrocytes. These data suggest that endothelial cells and astrocytes cooperate in the local synthesis and release of PUFA, collectively maintaining a brain environment enriched in long-chain PUFA. PMID- 11478375 TI - Lysophosphatidylcholine as a preferred carrier form of docosahexaenoic acid to the brain. AB - The metabolic fate of docosahexaenoic acid (DHA) was evaluated from its intake as a nutrient in triglycerides and phosphatidylcholines to its uptake by target tissues, especially the brain. Several approaches were used including the kinetics and tissue distribution of ingested 13C-labeled DHA, the incorporation of radiolabeled DHA injected as its nonesterified form compared to the fatty acid esterified in lysophosphatidylcholine (lysoPC), and the capacity of the two latter forms to cross a reconstituted blood-brain barrier (BBB) consisting of cocultures of brain-capillary endothelial cells and astrocytes. The results obtained allow us to raise the hypothesis that lysoPC may represent a preferred physiological carrier of DHA to the brain. PMID- 11478376 TI - Biosynthesis of docosahexaenoate-containing glycerolipid molecular species in the retina. AB - Vertebrate retinas are highly enriched in long-chain polyunsaturated fatty acids (PUFA), especially docosahexaenoic acid (22:6n-3, DHA). In the present study, we investigated the role of de novo synthesis in the enrichment of 22:6n-3 in characteristic molecular species of retinal glycerolipids. Following the incubation of fresh dark-adapted retinas with [2-(3)H]-glycerol, individual glycerolipids were isolated and converted into either diacylglycerol acetates (DGAC) or diacylglycerol benzoates (DGBZ), followed by high-performance liquid chromatography (HPLC) and flow-through radioactivity detection. Total lipids from rat retinas incubated with [3H]-glycerol were analyzed. Unlike what was observed with frog retinas, relative larger of amounts of di-22:6 molecular species were synthesized de novo. In both rat and frog retinas, there was synthesis of glycerolipid molecular species containing two PUFA (one of which was 22:6) in larger amounts than predicted by their steady-state mass levels. These results demonstrate that the unique molecular species of retinal glycerolipids are derived only in part through de novo synthesis, but that molecular rearrangement (remodeling) and differential turnover must also play a role in maintaining the high levels of 22:6 found in rod phohtoreceptor outer segments (ROS) membranes. PMID- 11478377 TI - Inhibition of neuronal apoptosis by polyunsaturated fatty acids. AB - The effect of polyunsaturated fatty acids (PUFAs), docosahexaenoic acid (22:6n-3; DHA) and arachidonic acid (20:4n-6; AA), on apoptotic cell death was evaluated based on DNA fragmentation and caspase-3 activity induced by serum starvation using Neuro-2A and PC-12 cells. The presence of 20:4n-6 in the medium during serum starvation decreased DNA fragmentation and this initial protective effect was diminished with prolonged serum starvation. The observed protective effect of 20:4n-6 was not affected by the inhibitors of cyclooxygenase (COX) and lipoxygenase. Conversely, 22:6n-3 became protective only after the enrichment of cells with this fatty acid at least for 24 h prior to the serum deprivation. DNA fragmentation as well as caspase-3 activity was reduced in 22:6n-3 enriched cells with a concomitant decrease in protein and mRNA levels. During the enrichment period, 22:6n-3 steadily increased its incorporation into PS leading to a significant increase in the total PS content; the protective effect of 22:6n-3 paralleled the PS accumulation. Neither direct exposure of cells to nor enrichment with 18:1n-9 had any protective effect. In conclusion, it is proposed that 20:4n-6 prevents neuronal apoptosis primarily due to the action of nonesterified 20:4n-6 but 22:6n-3, at least in part, through PS accumulation. PMID- 11478378 TI - Docosahexaenoic acid accumulation in the prenatal brain: prooxidant and antioxidant features. AB - Docosahexaenoic acid (DHA; 22:6n-3) is the major polyunsaturated fatty acid (FA) in the adult rat brain and it accumulates significantly more than any other FA prior to birth. Under normal nutritional conditions, fetal-brain DHA accumulation is substantial, with a "DHA accretion spurt" being demonstrated in the last period of gestation. Under stress conditions, this spurt may be harmful owing to an increase in multiple double-bond targets for lipid peroxidation. The "DHA accretion spurt" is supported by the maternal supply of DHA or its precursor. Under maternal dietary n-3 FA deficiency, DHA content in the fetal brain can be restored by direct intraamniotic injection of mM concentrations of ethyl-DHA (Et DHA). This approach may hold a potential advantage in the event of maternal-fetal insufficiency, a stress that may cause intrauterine growth retardation. It also revealed a potential beneficial effect after in utero ischemic stress; brain slices from Et-DHA-treated fetuses formed less oxidation products, as detected by thiobarbituric acid (TBA), compared to controls. Furthermore, brain-lipid extracts from Et-DHA but not ethyl-oleate treated fetuses, exhibited hydroxyl radical scavenging activity, as demonstrated by electron spin-resonance technique. Part of the beneficial effect of Et-DHA administration on the fetal brain may be attributed to enhanced free-radical scavenging capability, a phenomenon not directly related to vitamin E or lipid-soluble antioxidant levels. PMID- 11478379 TI - The role of docosahexaenoic acid containing phospholipids in modulating G protein coupled signaling pathways: visual transduction. AB - In order to understand the role of the high levels of docosahexaenoic acid (DHA) in neuronal and retinal tissue, a study of the effect of membrane lipid composition on the visual pathway, a G protein-coupled system, was undertaken. The level of metarhodopsin II (MII) formation was determined to be a function of phospholipid acyl-chain unsaturation, with the highest levels seen in DHA containing bilayers. Similarly, the rate of coupling of MII to the retinal G protein, Gt, to form a MII-Gt complex, was enhanced in DHA bilayers relative to less unsaturated phospholipids. Complex formation initiates the first stage of amplification in the visual pathway. The activation of the cGMP phosphodiesterase (PDE), the effector enzyme, represents the integrated pathway function. DHA containing bilayers were found to support PDE levels comparable to those of the rod outer segment (ROS) disk membranes. Inclusion of 30 mol% cholesterol in the reconstituted bilayers had an inhibitory effect on each step in the visual pathway studied. Inclusion of cholesterol reduced MII formation and PDE activity and increased the lag time between the appearance of MII and the formation of the MII-Gt complex. However, signaling in DHA bilayers was far less affected by the addition of cholesterol than in bilayers containing less unsaturated phospholipids. These studies point up the importance of DHA acyl chains in promoting optimal function in G protein-coupled signaling pathways. The results reported here suggest that visual and cognitive deficits observed in n-3 deficiency may result from decreased efficiency in related neurotransmitter and visual signaling pathways in the absence of DHA. PMID- 11478380 TI - In vivo fatty acid incorporation into brain phosholipids in relation to plasma availability, signal transduction and membrane remodeling. AB - A method, model, and "operational equations" are described to quantify in vivo turnover rates and half-lives of fatty acids within brain phospholipids, as well as rates of incorporation of these fatty acids into brain from plasma. In awake rats, recycling of fatty acids within brain phospholipids, due to deesterification and reesterification, is very rapid, with half-lives in some cases of minutes to hours. Plasma fatty acids make only a small contribution (2 4%) to the net quantity of fatty acids that are reesterified. This explains why many weeks are necessary to recover normal brain n-3 polyunsaturated fatty acid concentrations following their prolonged dietary deprivation. Changes in recycling of specific fatty acids in response to centrally acting drugs can help to identify enzyme targets for drug action. For example, recycling of arachidonate is specifically reduced by 80% in rats treated chronically with lithium, a drug effective against bipolar disorder; the effect reflects downregulation of gene expression of an arachidonate-specific phospholipase A2. When combined with neuroimaging (quantitative autoradiography in rodents or positron-emission tomography [PET] in macaques or humans), intravenously injected radiolabeled fatty acids can be used to localize and quantify brain PLA2-mediated signal transduction, and to examine neuroplastic remodeling of brain lipid membranes. PMID- 11478381 TI - Plasmalogens, phospholipase A2, and docosahexaenoic acid turnover in brain tissue. AB - Plasmalogens are glycerophospholipids of neural membranes containing vinyl ether bonds. Their synthetic pathway is located in peroxisomes and endoplasmic reticulum. The rate-limiting enzymes are in the peroxisomes and are induced by docosahexaenoic acid (DHA). Plasmalogens often contain arachidonic acid (AA) or DHA at the sn-2 position of the glycerol moiety. The receptor-mediated hydrolysis of plasmalogens by cytosolic plasmalogen-selective phospholipase A2 generates AA or DHA and lysoplasmalogens. AA is metabolized to eicosanoids. The mechanism of signaling with DHA is not known. The plasmalogen-selective phospholipase A2 differs from other intracellular phospholipases A2 in molecular mass, kinetic properties, substrate specificity, and response to glycosaminoglycans, gangliosides, and sialoglycoproteins. A major portion of [3H]DHA incorporated into neural membranes is found at the sn-2 position of ethanolamine glycerophospholipids. Studies with a mutant cell line defective in plasmalogen biosynthesis indicate that the incorporation of DHA is reduced in this RAW 264.7 cell line by 50%. In contrast, the incorporation of AA remains unaffected. This is reversed completely when the growth medium is supplemented with sn-1 hexadecylglycerol, suggesting that DHA can be selectively targeted for incorporation into plasmalogens. We suggest that deficiencies of DHA and plasmalogens in peroxisomal disorders, Alzheimer's disease (AD), depression, and attention deficit hyperactivity disorders (ADHD) may be responsible for abnormal signal transduction associated with learning disability, cognitive deficit, and visual dysfunction. These abnormalities in the signal-transduction process can be partially corrected by supplementation with a diet enriched with DHA. PMID- 11478382 TI - Polyunsaturated fatty acid regulation of gene expression. AB - Polyunsaturated fatty acids (PUFAs), specifically the n-3 and n-6 series, play a key role in the progression or prevention of human diseases such as obesity, diabetes, cancer, neurological and heart disease, mainly by affecting cellular membrane lipid composition, metabolism, signal-transduction pathways, and by direct control of gene expression. PUFAs show regulation of gene expression in several tissues, including brain, liver, heart, and adipose. Most recently, research has focused on identifying the mechanisms by which PUFAs regulate lipogenic gene expression. Research to date indicates that PUFA-mediated regulation of the genetic expression and proteolytic maturation of a group of transcription factors termed sterol regulatory element binding proteins (SREBPs) accounts for the suppression of hepatic lipogenic gene expression. However, our recent studies on the transcriptional regulation of the stearoyl-coenzyme A (CoA) desaturase gene, encoding a key enzyme in the cellular synthesis of monounsaturated fatty acids from saturated fatty acids indicates that PUFA can suppress gene transcription by a mechanism independent of SREBP maturation. PMID- 11478383 TI - Normal and defective neuronal membranes: structure and function: neuronal lesions in peroxisomal disorders. AB - Neuronal involvement in the peroxisomal disorders is divided into two main groups: developmental and postdevelopmental or degenerative. In the former the major lesions are neuronal migration abnormalities, which vary from severe in the cerebro-hepato-renal (Zellweger) syndrome (ZS) to mild in neonatal adrenoleukodystrophy. More common, but much less severe, are defects in neuronal differentiation or terminal migration, particularly involving the inferior medullary olives. Ultrastructural and neurochemical observations in ZS suggest that the presence of abnormal cytosomes in migrating neurons and radial glia, probably the result of excessive very long chain fatty acids, are responsible in part for its major neocortical migration defect, parasylvian pachygyria polymicrogyria. The postdevelopmental neuronal lesions involve specialized sensory neurons of the retina and the inner ear, resulting in atypical retinitis pigmentosa and its consequent visual defects and sensorineural hearing deficits. Neuronal atrophy and/or loss is seen in both the dorsal-root ganglia of adrenomyeloneuropathy and the atrophic cerebellum of rhizomelic chondodysplasia punctata. The underlying pathophysiology of these neuronal lesions is postulated to be caused by the incorporation of abnormal fatty acids into neuronal membranes, leading to an unresponsiveness to neurotrophic factors necessary for normal function and survival or to increased permeability of calcium channels and cell death. PMID- 11478384 TI - The peroxisome deficient PEX2 Zellweger mouse: pathologic and biochemical correlates of lipid dysfunction. AB - Zellweger syndrome is the prototypic human peroxisomal biogenesis disorder that results in abnormal neuronal migration in the central nervous system and severe neurologic dysfunction. A murine model for this disorder was previously developed by targeted deletion of the PEX2 peroxisomal gene. By labeling neuronal precursor cells in vivo with a mitotic marker, we can demonstrate a delay in neuronal migration in the cerebral cortex of homozygous PEX2 mutant mice. Postnatal PEX2 Zellweger mice develop severe cerebellar defects with abnormal Purkinje cell development and an altered folial pattern. When the PEX2 mutation is placed on an inbred murine genetic background, there is significant embryonic lethality and widespread neuronal lipidosis throughout the brain. Biochemical analysis of PEX2 mutant mice shows the characteristic accumulation of very long chain fatty acids and deficient plasmalogens in a wide variety of tissues. Docosahexaenoic acid levels (DHA; 22:6n-3) were found to be reduced in the brain of mutant mice but were normal in visceral organs at birth. All tissues examined in postnatal mutant mice had reduced DHA. The combined use of morphologic and biochemical analyses in these mice will be essential to elucidate the pathogenesis of this complex peroxisomal disease. PMID- 11478385 TI - Alterations in brain function after loss of docosahexaenoate due to dietary restriction of n-3 fatty acids. AB - The concentration of the major polyunsaturated fatty acid (PUFA) in brain, docosahexaenoate, may be markedly reduced by two or more generations of dietary restriction of sources of n-3 fatty acids in the diet. Such a deficiency was induced through the feeding of safflower oil as the principal source of essential fatty acids. The reference point for this diet was an n-3 adequate diet to which alpha-linoleate and docosahexaenoate were added through the addition of a small quantity of flax seed or algael oils, respectively. The loss of brain DHA was associated with poorer performance in spatial tasks and an olfactory-cued reversal learning task. No difference could be observed in the hippocampal gross morphology. This study demonstrates the importance of providing a source of n-3 fatty acids during mammalian growth and development. PMID- 11478386 TI - Restoring the DHA levels in the brains of Zellweger patients. AB - Patients with the Zellweger syndrome and its variants have very low levels of docosahexaenoic acid (DHA) in the brain, retina, and other tissues. Such a marked DHA deficiency could be related to the pathogenesis of peroxisomal disorders. Therefore, restoring the DHA levels in these patients can probably improve the clinical course of the disease. With this rationale, 20 patients with generalized peroxisomal disorders have been treated to date with DHA ethyl ester, at daily doses of 100-500 mg, for variable periods of time. Treatment has been always accompanied by a nutritious diet, normal for the age, in order to provide all the necessary nutrients and avoid a polyunsaturated fatty acid (PUFA) imbalance. The most constant improvement has been normalization of the DHA levels and liver function. Vision has improved in about half the patients and muscle tone has generally increased. Magnetic resonance imaging (MRI) examination revealed improvement of myelination in 9 patients. Significantly, the clinical improvement has been most marked in those patients who started the treatment before 6 mo of age. Biochemically, the plasma very long-chain fatty acids (VLCFA) 26:0 and 26:1n 9 decreased markedly despite the complete diet provided. In erythrocytes, the plasmalogen ratio 18: ODMA/18:0 increased in most cases, and sometimes even normalized. All these beneficial effects suggest that DHA deficiency plays a fundamental role in the pathogenesis of peroxisomal disease. Because DHA accretion is maximal during early brain development, it is essential to initiate the treatment as soon as possible. Otherwise, restoration of brain DHA levels and prevention of further damage will not be possible. PMID- 11478387 TI - Brain uptake and utilization of fatty acids: recommendations for future research. AB - A primary goal of the international workshop "Brain Uptake and Utilization of Fatty Acids" was to identify research areas that would benefit from further investigation. The major themes for future research are presented below: (1) Elucidating the role of the developing and mature cerebrovascular endothelium (CVE) in the uptake of fatty acids (FA) into the brain. (2) Clarifying the role of diffusion and receptor-mediated uptake of FAs by various brain cell membranes and protein-mediated shuttling of FAs between the CVE and various brain cells and tissues. (3) Illuminating the mechanisms of intermediate metabolism and the roles of polyunsaturated fatty acids (PUFA) in astrocytes, neurons and oligodendrocytes. Of special interest are the long-chain omega-3 PUFA and their derivatives, such as lipoproteins, phospholipids and plasmalogens, that have been associated with various disease states (such as those listed in [5], below). (4) Elucidating the role of gene expression on long-chain omega-3 PUFA incorporation in membranes and the regulatory role these and other PUFA have on gene expression in the brain. (5) Elucidating the recently identified roles of long-chain omega-3 PUFA in mood disorders, schizophrenia, stroke, peroxisomal biogenesis disorders, Huntington's disease, other neurodegenerative disorders and disorders of oxidative stress. (6) Undertaking placebo-controlled clinical trials to assess the therapeutic potential of omega-3 PUFA in the above disorders. (7) Developing new, and utilizing existing animal models in the above studies. (8) Developing noninvasive imaging and tagging methods for quantifying the migration and distribution of PUFA and their derivatives in the brain. (9) Applying multi disciplinary collaborations among biophysicists, physiologists and molecular biologists to the resolution of the above. PMID- 11478388 TI - Brain uptake and utilization of fatty acids: applications to peroxisomal biogenesis diseases. AB - The brain is rich in diverse fatty acids saturated, monounsaturated and polyunsaturated fatty acids with chain lengths ranging from less than 16 to more than 24 carbons that make up the complex lipids present in this organ. While some fatty acids are derived from endogenous synthesis, others must come from exogenous sources. The mechanism(s) by which fatty acids enter cells has been the subject of much debate. While some investigators argue for a protein-mediated process, others suggest that simple diffusion is sufficient. In the brain, uptake is further complicated by the presence of the blood-brain barrier. Brain fatty acid homeostasis is disturbed in many human disorders, as typified by the peroxisomal biogenesis diseases. A workshop designed to bring together researchers from varied backgrounds to discuss these issues in an open forum was held in March, 2000. In addition to assessing the current state of knowledge, areas requiring additional investigation were identified and recommendations for future research were made. A brief overview of the invited talks is presented here. PMID- 11478389 TI - Molecular architecture of the brain microvasculature: perspective on blood-brain transport. AB - Brain endothelial cells and their intercellular tight junctions form a cellular interface between the circulating blood and neural environment. All nutrients consumed by brain must traffic through this cellular space and its two limiting membranes. Additionally, the endothelial cell affects homeostasis by contributing or removing constituents from the interstitial space. These endothelial-cell functions are collectively accomplished with a rich complement of transporters and channels distributed, frequently asymmetrically, between the luminal and abluminal membranes. The identity and characterization of these proteins is rapidly advancing by application of molecular and cellular techniques. Knowledge of these molecular mechanisms will be beneficial in improving brain function and the treatment of neurological diseases. PMID- 11478390 TI - Fatty acid transport: the diffusion mechanism in model and biological membranes. AB - The transport of fatty acids (FA) across membranes can be described by three fundamental steps: adsorption, transmembrane movement, and desorption. In model membranes, these steps are all rapid and spontaneous for most fatty acids, suggesting that FA can enter cells by free diffusion rather than by protein mediated mechanisms. Here we present new fluorescence approaches that measure adsorption and transmembrane movement of FA independently. We show that FA adsorb to the plasma membrane of adipocytes and diffuse through the membrane by the flip flop mechanism within the time resolution of our measurements (approximately 5 s). Thus we show that passive diffusion is a viable mechanism, although we did not evaluate its exclusivity. Important implications of the diffusion mechanism for neural cells are that all types of FA could be available and that selectivity is controlled by metabolism. Studies of FA uptake into brain endothelial cells and other brain cell types need to be performed to determine mechanisms of uptake, and metabolism of FA must be separated in order to understand the role of membrane transport in the overall uptake process. PMID- 11478391 TI - Lipoxins and aspirin-triggered 15-epi-lipoxins are endogenous components of antiinflammation: emergence of the counterregulatory side. AB - Eicosanoids are known to play important roles in cell-cell communications and as intracellular signals that are critical components of multi-cellular responses such as acute inflammation and reperfusion injury. Recent findings have given rise to several new concepts that are reviewed here regarding the generation of eicosanoids and their impact in inflammation. Lipoxins (LX) are trihydroxytetraene-containing eicosanoids that can be generated within the vascular lumen during platelet-leukocyte interactions and at mucosal surfaces via leukocyte-epithelial cell interactions. During these cell-cell interactions, transcellular biosynthetic pathways are used as major LX biosynthetic routes, and thus, in humans, LX are formed in vivo during multi-cellular responses such as inflammation, atherosclerosis, and in asthma. This branch of the eicosanoid cascade generates specific tetraene-containing products that serve as stop signals, in that they regulate key steps in leukocyte trafficking and prevent leukocyte-mediated acute tissue injury. Of interest here are recent results indicating that aspirin's mechanism of action also involves the triggering of novel carbon 15 epimers of LX or 15-epi-LX that mimic the bioactions of native LX. Here, an overview of these recent developments is presented, with a focus on the cellular and molecular interactions of these novel antiinflammatory lipid mediators. PMID- 11478392 TI - Evasion of host immune surveillance by hepatitis C virus: potential roles in viral persistence. AB - Hepatitis C virus (HCV) is a major human pathogen that causes mild to severe liver disease worldwide. This positive strand RNA virus is remarkably efficient at establishing chronic infections. In order for a noncytopathic virus such as HCV to persist, the virus must escape immune recognition or evade host immune surveillance. Immune escape via the hypervariable region of the E2 envelope protein has been postulated as one mechanism for HCV persistent infection. Such hypervariability within the E2 protein may be under selective pressure from protective B cell or T cell responses and be able to escape immune recognition by rapid mutation of antigenic site. In addition to antigenic variation, HCV may also suppress immune response, leading to dampening of cellular immunity. This is supported by recent studies in our laboratory demonstrating that the HCV core protein can suppress host immune responses to vaccinia virus by downregulating viral specific cytotoxic T lymphocyte (CTL) responses and cytokine production. An understanding of the mechanisms behind HCV persistence will provide a basis for the rational design of vaccines and novel therapeutic agents targeting human HCV infection. PMID- 11478393 TI - Peptide loading of nascent MHC class I molecules. AB - A critical molecular interaction during assembly of the major histocompatibility complex (MHC) class I molecules takes place between the heavy chain and the transporter-associated with antigen-processing (TAP) complex. The recent mapping of regions of the heavy chain involved in the binding to TAP suggests a complex molecular interaction essential for the cell surface expression of the MHC class I. The advances made in understanding the TAP-MHC class I interaction are reviewed and discussed here. PMID- 11478394 TI - Human antibody expression in transgenic mice. AB - Human antibody repertoires can be created in transgenic mice following the introduction of human immunoglobulin heavy and light chain genes in their germline configuration. Transgene constructs or transloci have been obtained by plasmid assembly, cloning in yeast artificial chromosomes, and the use of chromosome fragments. Translocus integration and maintenance in transgenic mouse strains has been achieved by pronuclear DNA injection into oocytes and various transfection methods using embryonic stem cells. The human DNA segments rearrange faithfully in the mouse and produce extensive V(D)J combinations. Specific human monoclonal antibodies of high affinity for use in therapeutic applications have been produced from these translocus mice. PMID- 11478395 TI - The search for novel adjuvants for early life vaccinations: can "danger" motifs show us the way? AB - Potent but safe adjuvants are required to circumvent the many limitations of the newborn immune system to induce rapidly effective and long lasting immunity to subunit vaccines. By the use of pattern recognition receptors, antigen-presenting cells (APC) can very efficiently be activated by "danger" motifs expressed by various pathogens. APC activated by "danger" motifs, such as immunostimulatory sequences of bacterial DNA, can not only transmit the activation signal from the innate immunity to the adaptive compartment, but also shape the antigen-specific immune responses. Molecules or compounds expressing "danger" motifs could, therefore, be considered for use as adjuvants for subunit vaccines. In this review, the authors discuss the promises and potential drawbacks that such novel adjuvants could hold for their use in experimental and clinical early life vaccinations. PMID- 11478396 TI - CD89: the human myeloid IgA Fc receptor. AB - CD89 (Fc alphaRI) is the human myeloid IgA Fc receptor expressed on cells, such as neutrophils, eosinophils and monocytes/macrophages. Cross-linking of CD89 on these cells, by IgA-opsonised particles (e.g. bacteria, viruses) or anti-CD89 monoclonal antibodies, can trigger various immunological effector functions which are generally protective but may also cause harm to the body. CD89 is a transmembrane glycoprotein that binds both subclasses of IgA in all its molecular forms (i.e. monomeric, dimeric and secretory IgA) via a region of its membrane distal EC1 domain. DNA studies have shown that the CD89 gene is located within the newly described leukocyte receptor cluster (LRC) on chromosome 19. CD89 is more closely related to the KIR and MIR proteins, whose genes are also found in the LRC, than to other human Fc receptors (FcRs). On myeloid cells, CD89 is able to associate with the immunoreceptor tyrosine-based activation motif (ITAM) containing the FcR gamma chain, which is responsible for intracellular signaling via CD89. Recently, it has been suggested that some cells express CD89 in a form that does not associate with the FcR gamma chain. Although the biological relevance of this observation is not yet clear, it may explain certain anti inflammatory/inhibitory effects attributed to IgA. Here we review current knowledge concerning the genetics, structure and biological function of CD89. PMID- 11478397 TI - Immune-endocrine interactions of the hypothalamus-pituitary-thyroid axis: integration, communication and homeostasis. AB - The immune and neuroendocrine systems are two essential physiological components of mammalian organisms. Although each is primarily committed to a set of tasks involved, on the one hand, in the protection from infection and disease, and on the other hand, in the regulation of metabolism and other physiological activities, there is also evidence indicating that active and dynamic collaborations exist between those systems in the execution of their designated functions. These interactions occur at many stages of embryonic and neonatal development, and they are a continual part of the normal homeostatic balance needed to maintain health. The present review discusses various historical and contemporary perspectives of immune-endocrine interactions involving the hypothalamus-pituitary-thyroid axis, and offers a hypothesis of how this aspect of the neuroendocrine system participates directly in the immune response to antigenic challenge, infection and disease. PMID- 11478398 TI - Involvement of beta2-microglobulin in CD69 expression on T cells. AB - Beta2-microglobulin (beta2M) is the light chain of the class I HLA molecule. The serum level of beta2M is elevated in various diseases including lymphoma, inflammation, viral infections and chronic renal dysfunction. The present study addressed the possible influence of beta2M on T lymphocyte activation in vitro. Peripheral blood mononuclear cells from a group of 17 healthy subjects were examined. Stimulation with OKT3 and fibronectin in combination with 30 mg beta2M/dl resulted in a two-fold increase of cell proliferation. A similar effect was observed when OKT3 and collagen I were applied as well as when OKT3 and collagen IV were used as costimulation to T cells. The CD69 expression, measured by flow cytometry was significantly enhanced above the control level (1.52 +/- 1.03% vs 33.21 +/- 20.26%, p<0.01, control group and 30 mg beta2M/dl, respectively). Together, these observations suggest that beta2M may play a role in modulating lymphocyte proliferation, possibly through modification of the CD69 molecule. PMID- 11478399 TI - In vitro and in vivo study of the expression vector encoding vascular endothelial growth factor. AB - Vascular endothelial growth factor (VEGF) is an angiogenic cytokine with potential therapeutic applications in human diseases. It is a mitogen primarily for endothelial cells. The transfer of the cDNA encoding VEGF to ischemic tissues, which cannot be revascularized otherwise, represents a novel and promising approach to the treatment of vascular disorders. In this work the VEGF165 cDNA was cloned into the expression vector pSecTag2B. The activity of the construct was studied in cell culture as well as in vivo. Western blotting study showed that the cells transfected with the vector secreted significantly higher amounts of VEGF to the culture medium than the non-transfected cells. In vivo study revealed an increased number of new vessels in animals injected with vector encoding VEGF as compared with empty plasmid. Also, tumor cells transfected with the VEGF plasmid exhibited extensive vascularization. PMID- 11478400 TI - Modification of collagen film by certain chemical agents. AB - Morphological changes and the content of free carboxyl groups in bovine collagen (type I) film under the influence of trypsin, hydrochloric acid (HCl) and ethylenediaminetetraacetic acid (EDTA) were studied. Incubation with trypsin and HCl was found to cause some delamination of the film and the appearance of some low-density spots. Incubation with EDTA did not cause any morphological changes. A high concentration of free carboxyl groups (10-fold higher than in control) was seen after incubation with trypsin. PMID- 11478401 TI - In vitro culture of human epithelial cells on a modified xenogenic collagen support. AB - Human epithelial cells (HeLa, HaCaT, NHK) were cultured in vitro on chemically modified collagen membranes. Adhesion to the support was measured by estimation of the percentage of adhering 51Cr-labeled cells. Proliferation was estimated with the XTT test. Morphological observations of cells growing on HCl-treated collagen were performed using histological and electron microscopic techniques. HCl and trypsin-modified xenogenic collagen was found to be a good support for human cells in vitro. EDTA-incubated collagen enhanced neither adhesion nor proliferation. The best adhesion and proliferation were found on HCl-treated collagen, depending, however, on the kind of cells. PMID- 11478403 TI - Plans for PubMed Express scrapped. PMID- 11478402 TI - Tension mounts over the future of stem cell research. PMID- 11478404 TI - NBAC report calls for increased federal oversight of research on human subjects. PMID- 11478405 TI - Letters to Kirschstein voice concerns over implementation of loan repayment program. PMID- 11478406 TI - Obesity is associated with impaired ventricular protein kinase C-MAP kinase signaling and altered ANP mRNA expression in the heart of adult Zucker rats. AB - BACKGROUND: In the obesity model of the Zucker rat, myocardial protein kinase C (PKC) activation by phorbol ester is impaired. The influence of obesity on myocardial cell signaling was investigated by studying the activation of PKC isozymes and MAP kinases (MAPK) p38 and p42/44 as well as the induction of ANP mRNA. METHODS: Isolated hearts obtained from 17-week-old lean and obese Zucker rats were perfused with 200 nM phorbol 12-myristate 13-acetate (PMA) at different time periods. Immunodetectable PKC isozymes, phosphorylated-MAPK, and ANP mRNA were determined by Western and Northern blots, respectively. RESULTS: PMA promoted a marked transient translocation of ventricular PKCalpha from the cytosol to the membranes within 10 minutes in lean rats, whereas it had a much weaker effect in obese rats. Moreover, PMA induced a significant activation of PKCdelta in lean but not in obese rat hearts. After PKC activation, increases in phosphorylation levels of myocardial p38 and p42 MAPK were approximately 3-fold higher in lean rats than in obese animals. Concerning the induction of ANP, PMA transiently tripled ANP mRNA within 60 minutes in lean but not in obese rats. CONCLUSIONS: In the genetically obese Zucker rat, the myocardial signal transduction cascade PKC-MAPK-ANP mRNA seems to be markedly impaired. It can be speculated that this abnormal cardiac cell signaling in obese rats reflects an early phase in the cardiac pathogenesis accompanying obesity. PMID- 11478407 TI - Effect of simvastatin alone and in combination with cytosine arabinoside on the proliferation of myeloid leukemia cell lines. AB - BACKGROUND: Cholesterol biosynthesis is regulated by the activity of 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase. Cholesterol and its derivatives are required in high concentrations by neoplastic proliferating cells for both DNA synthesis and cell growth. Thus, inhibition of HMG-CoA reductase could effect cell cycle progression and proliferation. Therefore, we examined the effect of an HMG-CoA reductase inhibitor (simvastatin) alone and in combination with cytosine arabinoside (ARA-C) on the proliferation of two AML cell lines. METHODS: AML blasts derived from two cell lines (HL-60 and AML-2) were incubated with increasing concentrations of either simvastatin alone or simvastatin alone for 24 hours with ARA-C added thereafter. The effect of the drugs on cell proliferation in liquid culture (3H thymidine uptake) and on clonogenic assay was analyzed. RESULTS: We found that the number of proliferating AML blasts (suspension cultures) and colony formations (agar cultures) of both cell lines declined significantly after incubation with simvastatin. Preincubation of both cell lines with simvastatin by the addition of increasing concentrations of ARA-C produced a degree of growth inhibition that was significantly greater than that of the individual compounds. This antigrowth interaction was additive rather than synergistic. CONCLUSIONS: We conclude that simvastatin has a major antiproliferative effect on AML blasts in vitro. Also, the combination of simvastatin and ARA-C significantly enhanced the antiproliferative effect of each drug. These findings may open new avenues in both the laboratory and clinical research of the treatment of leukemia. PMID- 11478408 TI - Basal and post-methionine serum homocysteine and lipoprotein abnormalities in patients with chronic liver disease. AB - BACKGROUND: Lipoprotein abnormalities are commonly found in chronic liver diseases (CLDs), particularly hypercholesterolemia in primary biliary cirrhosis (PBC). However, affected patients may not be at increased risk of coronary heart disease. Cirrhotic patients display impaired methionine clearance, and an increased level of homocysteine, a methionine metabolite, is an independent risk factor for coronary heart disease. Thus, we hypothesized that the low risk of coronary heart disease in patients with CLD may be related to low serum levels of homocysteine. The aim of this study was to test this hypothesis after methionine load and to describe the serum lipoprotein profile in patients with PBC and in patients with hepatocellular liver disease. METHODS: Fifteen female patients (mean age, 58.2 +/- 11.7 years) with PBC, 15 female patients (mean age, 54.5 +/- 9.6 years) with other causes of CLD, and 15 healthy sex- and age-matched controls were given L-methionine (50 mg/kg of ideal body weight). Basal fasting serum homocysteine level and 2, 4, and 6 hours of post-methionine load were determined using high-performance liquid chromatography with a fluorometric detector. Levels of fasting serum cholesterol, triglycerides, high-density lipoprotein (HDL), low density lipoprotein (LDL), lipoprotein (a) (Lp(a)), and apoprotein B were also determined. RESULTS: Results showed that mean basal and post-methionine load (6 hours) serum homocysteine levels were statistically significantly higher in the patients with PBC and with CLD than in the control group (P=0.04) and that levels of serum cholesterol, LDL, HDL, and apoprotein B were significantly higher in the PBC patients than in the other two groups (P < or = 0.05). There was no correlation between any of these parameters and the severity of liver disease. Serum HDL was significantly lower in the CLD group (P < or = 0.05) and correlated with severity of liver disease. There was no significant difference in serum cholesterol, LDL, or apoprotein B between the CLD group and the controls. Serum triglyceride and Lp(a) levels were similar for all three groups. CONCLUSIONS: In contrast to previous reports, the site of the methionine metabolic impairment was found to be below the homocysteine synthesis level. For most patients with CLD, factors other than serum homocysteine or Lp(a) are responsible for the reduction in the risk of coronary heart disease. Further studies with larger samples are needed. PMID- 11478409 TI - Effect of dexamethasone on TSH secretion induced by TRH in human obesity. AB - BACKGROUND: The presence of an abnormally high thyroid-stimulating hormone (TSH) response to thyrotropin-releasing hormone (TRH) makes it difficult to distinguish some euthyroid obese subjects from subelinically hypothyroid obese patients. Here, we examine whether such distinction may be achieved after treatment with glucocorticoids, which inhibit TSH secretion at the hypothalamic-pituitary level. METHODS: TRH tests (200 microg as an intravenous bolus injection) were performed in 30 age- and weight-matched, obese, but otherwise healthy, men. All subjects were tested again with TRH after treatment with dexamethasone (dex) (2 mg/d in four divided doses orally for 3 days). RESULTS: In all subjects, total thyroxine and triiodothyronine concentrations were in the normal range. According to basal and TRH-stimulated serum thyrotropin (TSH) levels, subjects were divided into the following three groups: group I (n=10), euthyroid subjects; group II (n=10), euthyroid subjects with normal basal but abnormally elevated TSH responses to TRH; group III (n=10), subjects with elevated basal and TRH-induced TSH levels (subclinical hypothyroidism). Basal TSH levels were 1.8+/-0.4 mU/L in group I, 1.7+/-0.3 in group II, and 6.0+/-0.7 in group III. In both groups II and III, TRH induced TSH increments were above the normal range (maximal increment> 15 mU/L) and were significantly higher than in group I. After the second treatment with TRH, pretreatment with dex significantly decreased both basal TSH levels and peak TSH responses to TRH in all groups. However, a striking percentage decrease (>50%) in TRH-induced peak TSH responses was observed in euthyroid obese subjects of groups I and II, whereas hypothyroid subjects of group III showed only a slight decrement (<25%). CONCLUSIONS: The sensitivity of the TSH secretory system to glucocorticoid inhibitory action is preserved in obese subjects with abnormally elevated TSH response to TRH, but not in subclinically hypothyroid obese patients. The TRH plus dex test might be useful in future studies to understand the mechanisms underlying alterations in TSH secretion in obesity. PMID- 11478410 TI - Erythropoietin withdrawal alters interactions between young red blood cells, splenic endothelial cells, and macrophages: an in vitro model of neocytolysis. AB - BACKGROUND: We have described the rapid destruction of young red blood cells (neocytolysis) in astronauts adapting to microgravity, in polycythemic high altitude dwellers who descend to sea level, and in patients with kidney disorders. This destruction results from a decrease in erythropoietin (EPO) production. We hypothesized that such EPO withdrawal could trigger physiological changes in cells other than red cell precursors and possibly lead to the uptake and destruction of young red cells by altering endothelial cell-macrophage interactions, most likely occurring in the spleen. METHODS: We identified EPO receptors on human splenic endothelial cells (HSEC) and investigated the responses of these cells to EPO withdrawal. RESULTS: A monolayer of HSEC, unlike human endothelial cells from aorta, glomerulus, or umbilical vein, demonstrated an increase in permeability upon EPO withdrawal that was accompanied by unique morphological changes. When HSEC were cultured with monocyte-derived macrophages (but not when either cell type was cultured alone), EPO withdrawal induced an increased ingestion of young red cells by macrophages when compared with the constant presence or absence of EPO. CONCLUSIONS: HSEC may represent a unique cell type that is able to respond to EPO withdrawal by increasing permeability and interacting with phagocytic macrophages, which leads to neocytolysis. PMID- 11478411 TI - Inhibition of L-type Ca2+ channel current in Xenopus oocytes by amiodarone. AB - BACKGROUND: Although amiodarone has been referred to as a class III antiarrhythmic agent, it also possesses electrophysiologic characteristics of the three other classes (classes I and IV and minor class II effects). Previous studies have demonstrated that amiodarone inhibits Ca2+ channel current in intact cardiac myocytes. However, it is not clear whether this response reflects a pure class IV effect (direct Ca2+ channel inhibition) or a class II effect (beta adrenergic receptor blockade) of amiodarone. METHODS: In the current study, the effects of amiodarone on Ca2+ current were studied in the absence of sympathetic regulation using a Xenopus oocyte expression system. The L-type Ca2+ channel alpha1C subunit was coexpressed with the alpha2delta and beta2a subunits in enzymatically digested Xenopus oocytes. Ca2+ currents were recorded using the cut open oocyte preparation. RESULTS: We found that perfusion of 10 microM isoproterenol produced no significant change in peak Ca2+ current (from 223+/-33 to 210+/-29 nA, mean+/-SEM, n=5, P=not significant), indicating the absence of a functional stimulatory sympathetic signal pathway in these oocytes. After 10 minutes of exposure to 10 microM amiodarone, Ca2+ current amplitude was significantly decreased from 174+/-33 to 100+/-26 nA (n=8, P<0.01; control group: 220+/-33 to 212+/-29 nA, n=5, P=not significant). These effects were similar to those of 10 microM nifedipine (201+/-48 to 108+/-48 nA, n=6, P<0.05), a typical Ca2+ channel blocker. On the other hand, neither amiodarone nor nifedipine significantly altered the Ca2+ current activation or inactivation kinetics. CONCLUSIONS: These results demonstrate that amiodarone inhibits Ca2+ current in the absence of a functional intrinsic beta-adrenergic stimulatory system and, therefore, represents a true class IV effect. PMID- 11478412 TI - Association of traditional risk factors with coronary calcification in persons with a family history of premature coronary heart disease: the study of the inherited risk of coronary atherosclerosis. AB - BACKGROUND: Genetic factors strongly influence the risk of coronary heart disease (CHD), but their contribution to variation in coronary atherosclerosis beyond that measured by traditional CHD risk factors is uncertain. METHODS: We recruited healthy subjects with family histories of premature CHD. We assessed traditional risk factors and performed electron beam tomography (EBT) to quantitate coronary artery calcification (CAC), a marker of coronary atherosclerosis. Persons with significant risk factors that included diabetes, total cholesterol >300 mg/dL, active cigarette smoking, and poorly controlled hypertension were excluded from the study. In this paper, we report on the relationship between traditional risk factors and CAC in this cohort. RESULTS: The incidence of coronary calcification was significantly higher in this cohort than in the population-based Rochester Heart Study. In our cohort, most traditional risk factors were significantly associated with CAC on univariate analysis. On the other hand, in stepwise logistical regression, age and triglycerides were the only predictors of variation in CAC in men and accounted for only 30% of the variation; in women, age, body mass index (BMI), and triglycerides were the only traditional risk factors significantly associated with CAC variation and accounted for 22.2% of CAC variance. CONCLUSIONS: In a cohort of subjects specifically selected for the characteristic of a family history of premature CHD, traditional risk factors accounted for less than one-third of the variation in CAC, and the most important predictors of CAC after age were plasma triglycerides. This supports the opinion that other inherited risk factors have important effects on the variation in coronary atherosclerosis and that the strategy of using EBT to phenotype clinically asymptomatic subjects with regard to coronary atherosclerosis may be a useful tool for identification of genes that are associated with CHD. PMID- 11478413 TI - Monocyte chemoattractant protein-1 and interleukin-8 are increased in bronchopulmonary dysplasia: relation to isolation of Ureaplasma urealyticum. AB - BACKGROUND: An exaggerated inflammatory response occurs in infants who subsequently develop bronchopulmonary dysplasia (BPD). Ureaplasma urealyticum (Uu) is frequently isolated from cultures of tracheal secretions obtained from very low birth weight infants and is associated with an increased risk of BPD. METHODS: We examined the relationships between isolation of genital mycoplasmas, tracheal aspirate (TA) interleukin-8 (IL-8), and monocyte chemoattractant protein 1 (MCP-1) concentrations and the development of BPD. Serial TAs were obtained prospectively from 35 very low birth weight infants, and IL-8 and MCP-1 concentrations were determined by enzyme-linked immunoadsorbent assay. Tracheal cultures for bacteria and genital mycoplasmas were performed on aspirates obtained during the first 2 days of life. RESULTS: Infants who developed BPD (n=18) were less mature (25.2+/-0.2 vs 27.8+/-0.5 weeks; P<0.001), of lower birth weight (746+/-28 vs 1052+/-41 g; P<0.001), and more likely to have a positive tracheal culture for Uu (39% vs 6%; P=0.026) than those who did not develop BPD (n=17). Tracheal concentrations of IL-8 and MCP-1 were significantly increased in infants who developed BPD (IL-8: P=0.0001; MCP-1: P<0.001, analysis of variance) and correlated with duration of mechanical ventilation and oxygen treatment. Uu positive infants had an increased incidence of BPD (88% in infants with Uu vs 42% in infants without Uu; P=0.020) and had TA concentrations of IL-8 and MCP-1 that were significantly increased compared with those of Uu-negative infants. CONCLUSIONS: Increased TA concentrations of IL-8 and MCP-1 during the first 2 weeks of life are associated with the development of BPD. Recovery of Uu from TAs is associated with a more robust inflammatory reaction and an increased risk of BPD. PMID- 11478414 TI - Two steps forward, one step back; paradigmatic changes in psychiatry. AB - The initial scientification of psychiatry after the European discovery of psychotropic drugs via psychopharmacology and biological psychiatry has gradually ended in an impasse. As a result of at least four developments, psychiatry is re orientating in a more dimensional and functional direction in order to adapt to the progress in the neurosciences. Rigid adherence to nosological systems should be replaced by a careful observation and description of psychopathology as well as a search for relevant psychological dysfunctions and their putative pathophysiological basis. Psychotropics have to be developed that do not suppress symptoms but promote potentially intact functions. Emphasis should be given to the brain mechanisms that are involved in habit formation and chronicity. Modern genetics will drive the discipline towards early intervention and prevention and will elucidate the relations between genetic disorders and the behavioural phenotype. The basis, however, remains clinical observation, dynamically reshaped by neuroscience. PMID- 11478415 TI - Validity and reliability of leonhard's classification of endogenous psychoses: preliminary report on a prospective 25- to 30-year follow-up study. AB - A 25- to 30-year controlled follow-up investigation of endogenous psychoses started in 1997. The research program labeled "Budapest 2000" was initiated in 1967 as a controlled prospective study. The assessment of 108 patients and 24 normal control persons has so far been completed. With regard to the "middle groups" in the Leonhardian classification, diagnoses of bipolar manic-depressive psychosis, cycloid psychosis, periodic catatonia, systematic paraphrenia, systematic catatonia and hebephrenia proved to be valid in the long term. Revision of the category affect-laden paraphrenia seems to be called for. PMID- 11478416 TI - Catatonia: syndrome or schizophrenia subtype? Recognition and treatment. AB - Two views of catatonia influence clinical practice. In the classical European view, adopted by DSM classifications, the signs of catatonia indicate a form of schizophrenia. In the syndromal view, the signs of catatonia are motor signs that are readily identified in many psychiatric disorders. Catatonia is a parallel behavior phenomenon to delusions (in thought) and delirium (in cognition). The syndromic view includes the neuroleptic malignant syndrome. It encourages a different treatment algorithm, the use of benzodiazepines and electroconvulsive therapy, to replace the customary use of antipsychotic drugs alone. The benefits of such treatment warrant the recommended change in concept and classification. PMID- 11478417 TI - Increased volume of the nucleus accumbens in schizophrenia. AB - The nucleus accumbens, an integral and important part of limbic and prefrontal cortico-striato-pallidal-thalamic circuits, is involved in several cognitive, emotional and psychomotor functions altered in schizophrenia. In animal models, developmental disturbances within the entorhinal cortex and the hippocampus induce a dysregulation of inputs to the nucleus accumbens resulting in behavioral abnormalities which point to psychotic psychopathology. Nonetheless, due to the complex neuroanatomy of the human ventral striatum hardly any morphometric data on the nucleus accumbens are available. A postmortem stereological investigation of the nucleus accumbens was performed in complete brains of 9 male schizophrenics and 9 male controls between the ages of 46 and 64. Complete serial coronal slices of both hemispheres were stained with a modified Nissl-technique. Tissue shrinkage after staining and embedding was corrected for both individual and regional shrinkage. Based on recent precise delimitations of the human ventral striatum, in vivo hemisphere-adjusted volumes of the nucleus accumbens (volume densities) and absolute accumbal volumes were calculated applying the Cavalieri-estimator. In schizophrenics, mean hemisphere-adjusted volumes of the nucleus accumbens were significantly increased on both sides (right accumbens: p = 0,005**; left accumbens: p = 0,016*). Hemispherical volumes and volumes of the nucleus accumbens were significantly correlated in both groups (p = 0,02*). Most likely, this increase in volume of the ventral striatum reflects a decrease in naturally occuring cell death following prenatal cortical neurodevelopmental disturbances. PMID- 11478418 TI - Disturbed neural circuits in a subtype of chronic catatonic schizophrenia demonstrated by F-18-FDG-PET and F-18-DOPA-PET. AB - Permanent verbal, visual scenic and coenaestetic hallucinations are the most prominent psychopathological symptoms aside from psychomotor disorders in speech sluggish catatonia, a subtype of chronic catatonic schizophrenia according to Karl Leonhard. These continuous hallucinations serve as an excellent paradigm for the investigation of the assumed functional disturbances of cortical circuits in schizophrenia. Data from positron emission tomography (F-18-FDG-PET and F-18-DOPA PET) from three patients with this rare phenotype were available (two cases of simple speech-sluggish catatonia, one case of a combined speech-prompt/speech sluggish subtype) and were compared with a control collective. During their permanent hallucinations, all catatonic patients showed a clear bitemporal hypometabolism in the F-18-FDG-PET. Both patients with the simple speech-sluggish catatonia showed an additional bilateral thalamic hypermetabolism and an additional bilateral hypometabolism of the frontal cortex, especially on the left side. In contrast, the patient with the combined speech-prompt/speech-sluggish catatonia showed a bilateral thalamic hypometabolism combined with a bifrontal cortical hypermetabolism. However, the left/right ratio of the frontal cortex also showed a lateralisation effect with a clear relative hypometabolism of the left frontal cortex. The F-18-DOPA-PET of both schizophrenic patients with simple speech-sluggish catatonia showed a normal F-18-DOPA storage in the striatum, whereas in the right putamen of the patient with the combined form a higher right/left ratio in F-DOPA storage was discernible, indicating an additional lateralized influence of the dopaminergic system in this subtype of chronic catatonic schizophrenia. Most likely, the prominent bitemporal F-18-FDG- hypometabolism in these chronic schizophrenic patients with speech-sluggish catatonia suffering from permanent continuous hallucinations, reflects a deficit in sensoric gating following prenatal cortical neurodevelopmental disturbances. However, the functional disturbances underlying hallucinations in "the schizophrenias" seem to be more complex; in different subtypes of the schizophrenic spectrum disorder hallucinations seem to be based on alterations in additional cortical and subcortical brain regions. PMID- 11478419 TI - Dopamine D3 receptor gene and tardive dyskinesia in Chinese schizophrenic patients. AB - Epidemiological studies have shown a lower prevalence of tardive dyskinesia (TD) among Chinese psychiatric patients compared to Caucasian and Black patient populations. It has been hypothesized that pharmacogenetic factors may underlie this cross-cultural difference. Due to the important implications of the dopamine D3 receptor gene (DRD3) in motor control, we investigated the frequency of polymorphic serine (ser) to glycine (gly) substitution of the gene DRD3 in Chinese schizophrenic patients. The sample size consisted of 65 patients with TD and 66 without TD. Patients were assessed for the severity of TD, the presence of akathisia and parkinsonian symptoms and were subsequently genotyped. We found no evidence that the dopamine D3 receptor gene is likely to confer susceptibility to the development of tardive dyskinesia in Chinese patients with schizophrenia. PMID- 11478420 TI - Electrophysiology of the prefrontal cortex in healthy controls and schizophrenic patients: a review. AB - Prefrontal brain regions, in particular the anterior cingular gyrus (ACG), are altered in chronic schizophrenic patients and also play a pivotal role in physiological mechanisms of response control. A cued Continuous Performance Test (CPT) is a suitable paradigm for the investigation of response control, as it contains its fundamental mechanisms, i.e. the execution (Go) and the inhibition (NoGo) of an anticipated motor response. Previous electrophysiological investigations in healthy subjects during CPT execution revealed that the gravity center (centroid) of the event-related potential (ERP) elicited during the NoGo condition is located more anterior as compared to the Go-condition in every single case. Moreover, by means of an electrophysiological source location method this NoGo-anteriorisation phenomenon (NGA) has been attributed to a strong brain electrical NoGo-hyperactivity located in prefrontal brain areas (ACG). Furthermore, the NGA has a superior test-retest reliability, a very high interindividual stability and is independent from age- and gender-effects. Systematic schizophrenic patients were characterized by a significantly diminished NGA indicating a dysfunction of prefrontal brain areas including the ACG. Based on these findings, the NGA has to be considered as a first electrophysiological measure for the quantitative assessment of ACG-function within a response control paradigm. Hopefully, this biological parameter will contribute to the elucidation of the etiopathogenetical background in different schizophrenic diseases. PMID- 11478421 TI - Magnetic resonance imaging findings in schizophrenia and atypical psychoses. AB - The differences among MRI findings were studied in schizophrenic psychoses. The schizophrenics and atypical psychotics had significant reductions in bilateral hippocampal volumes compared to controls, but the two patient groups did not differ from each other. As for ventricle volume, the schizophrenics showed significantly larger temporal horns and third ventricle than normal controls, whereas atypical psychotics did not. Moreover, the left temporal horn in the schizophrenics was significantly larger than that seen in the atypical psychotics. By cluster analysis, schizophrenics and atypical psychotics were found to have a tendency to be distributed in different groups. These results might be considered to support the classification of schizophrenic psychoses into schizophrenia and atypical psychoses. PMID- 11478422 TI - Pharmacotherapy of depression: a historical analysis. AB - Iproniazid and imipramine, the prototypes of monoamine oxidase inhibitor (MAOI) and monoamine (re)uptake inhibitor (MAUI) antidepressants, were introduced in 1957. The relationship between iproniazid's antidepressant effect and its MAO inhibiting property was tenuous. Because of the potential drug-drug interactions and the need for dietary restrictions, the use of MAOIs became restricted to atypical depression. The confounding of reserpine reversal with antidepressant effect led to the theory that MAU inhibition is responsible for imipramine's antidepressant effect. Driven by neuropharmacological theory, non-selective reuptake inhibitors were replaced first by selective norepinephrine reuptake inhibitors, then by selective serotonin reuptake inhibitors, and more recently, by a series of new antidepressants to relieve the stimulation of serotonin-5HT2A receptors and the compensatory decline of dopamine in the brain. Each antidepressant has its own identity, but meta-analyses indicate a widening of the antidepressant response range from 65-70% to 45-79%, and a lowering of the antidepressant threshold from 65% to 45%. Although one can no longer expect that 2 of 3 depressed patients will respond to treatment, the newer antidepressants are better tolerated, because they produce less anticholinergic side effects. PMID- 11478423 TI - Molecular foundation of anxiety disorders. AB - Genetic epidemiology has assembled convincing evidence that anxiety and related disorders are influenced by genetic factors and that the genetic component is highly complex, polygenic, and epistatic. Although several genes which may contribute to the genetic variance of anxiety-related traits or modify the phenotypic expression of pathologic anxiety are currently under investigation, molecular genetics has so far failed to identify a genomic variation that can consistently contribute susceptibility of anxiety disorders. Investigation of gene-gene and gene-environment interactions in humans and nonhuman primates as well as gene inactivation studies in mice further intensify the identification of genes that are essential for development and adult plasticity of the brain related to complex anxiety responses. Because the modes of inheritance of anxiety disorders are complex, it has been concluded that multiple genes of small effect, in interaction with each other and with nongenetic neurodevelopmental events, produce vulnerability to the disorder. Future research directions will take advantage of the completion of the sequencing the human and mouse genome coinciding with the revolution in bioinformatics. More than 1.4 million single nucleotide polymorphisms (SNPs) in the human genome have been identified. This collection should allow the initiation of genome-wide linkage disequilibrium mapping of the genes influencing anxiety in the human population. Integration of these emerging tools and technologies for genetic analysis will provide the groundwork for an advanced stage of gene identification and functional studies in anxiety and related disorders. PMID- 11478424 TI - Naturally occurring benzodiazepines may codetermine chronotypes. AB - Determinants of individual differences in sleep-wake cycles and vigilance are being recognized as major factors of influence in both physical and mental health. Alterations of an accustomed circadian sleep-wake rhythm are commonly seen in the early stages of the majority of psychiatric disorders and, by themselves, predispose to significant morbidity even in the absence of an underlying illness. While it is well known that disruptions of sleep respond favourably to benzodiazepines, agents which have been prescribed for insomnia since their industrial synthesis in the early 1960s, little attention has been paid to putative central nervous system effects of naturally occurring benzodiazepines. These molecules were found in various nutritive plants and have been quantified in human brain and peripheral blood of drug-naive individuals at trace amounts. There is agreement that elevated concentrations of naturally occurring benzodiazepines participate in the complex pathophysiology of hepatic encephalopathy, a condition asssociated with progressive impairment of consciousness and, ultimately, coma. In the present study, we address the relationship between naturally occurring benzodiazepines and time-of-day effects on the behavior of healthy subjects. PMID- 11478425 TI - Neurobiology of an addiction memory. AB - The existence of an "addiction memory" (AM) and its importance in relapse occurrence and maintenance of learned addictive behaviour will be explained with neurobiological and clinical arguments. Because the human brain is an open learning system, which reveals its own neuronal connectivity through the experience of the perceived environment with its own state, the personal AM is interpreted as an individual acquired software disturbance in relation to selectively integrating "feedback loops" and "comparator systems" of neuronal information processing. This is in accordance with the experience that the AM and its specific cue reactivity can be activated at any time by relapse-endangering complex internal and/or external situations with cue stimulated craving. The AM becomes part of the personality represented on the molecular level via the neuronal level and the neuropsychological level, especially in the episodic memory. This neurobiological unchangeable imprinted addictive behaviour with "loss of control" and "obsessive-compulsive craving" was also found in a long term learning model with rats (Wolffgramm). Identical homological phylogenetically old brain structures for learning mechanisms allow the comparison between human and animal behaviour. The AM seems to be a clinical empirical proved reality. It is compatible with recently discussed results of neurosciences. PMID- 11478426 TI - Neurotransmission in HIV associated dementia: a short review. AB - Human immunodeficiency virus (HIV) infection is frequently associated with specific neurological and psychiatric symptoms. Our understanding of how HIV related CNS deficits develop is still preliminary and the cause remains obscure. However, some clues have emerged which may clarify uncertainties. Following a brief discussion of the epidemiology underlying neuropathological mechanisms and clinical symptoms in HIV-infected patients, we focus our attention on neurochemical data obtained by studies in humans and rhesus monkeys which provide information on the effect of the retroviral infection on neurotransmission and assist in the evaluation of potential therapeutic treatments. PMID- 11478427 TI - Increased blood pressure and loss of anp-induced natriuresis in mice lacking DARPP-32 gene. AB - Atrial natriuretic peptide (ANP) is an important regulator of sodium metabolism and indirectly of blood pressure. Evidence has accumulated that ANP regulates sodium metabolism through a cascade of steps involving an increase in the level of cGMP, activation of cGMP-dependent protein kinase (PKG), and inhibition of renal tubular Na+, K+-ATPase activity. One of the major substrates for PKG is DARPP-32. In the present study we observed that ANP does not induce natriuresis in mice that lack DARPP-32. In contrast, there was a 4-fold increase in urinary sodium excretion following ANP administration to wild type mice. ANP as well as Zaprinast, a selective inhibitor of cGMP phosophodiesterase, inhibited renal Na+, K+-ATPase activity in wild type mice but had no such effect in mice lacking DARPP 32. Mean arterial blood pressure, measured in conscious animals, was significantly increased in DARPP-32 deficient mice as compared to wild type mice. The results confirm that DARPP-32 acts as a third messenger in the ANP signaling pathway in renal tissue and suggest an important role of DARPP-32 in the maintenance of normal blood pressure. PMID- 11478428 TI - Acute abdominal symptoms in malignant hypertension: clinical presentation in five cases. AB - Malignant hypertension causes anatomical and functional damage in several target organs, in particular brain, retina, heart and kidneys. Although vascular lesions in the gastroenteric tract are known to occur in several instances, their clinical relevance is unknown. In this study five cases of malignant hypertension, presenting with acute abdominal symptoms, are reported. A history of essential arterial hypertension was present in three patients; while one patient had a previous diagnosis of renovascular hypertension and one patient had renoparenchymal hypertension. However, in all cases the antihypertensive treatment was discontinued and inadequate before the accelerated malignant phase. The acute abdominal symptoms at presentation were due to intestinal infarction in 3 patients and acute pancreatitis in 2 patients. One patient with intestinal infarction died of postoperative cardiogenic shock. Our data are in agreement with previous reports describing the possible intra-abdominal complications of malignant hypertension. The therapeutic approach in such conditions should always consider an effective antihypertensive treatment in conjunction with surgical options. PMID- 11478429 TI - A family with liddle's syndrome caused by a mutation in the beta subunit of the epithelial sodium channel. AB - Liddle's syndrome is a rare form of autosomal-dominant salt-sensitive hypertension. Constitutive activation of the amiloride-sensitive distal renal epithelial sodium channel (ENaC) is essential for salt-sensitive hypertension. Recently, several DNA analysis studies have indicated that there is a mutation of C-terminus of either the beta or y subunit. We sequenced the C-termini of the beta and -gamma subunits of the ENaC in a Japanese family with hypertension and hypopotassemia without excess minerarocorticoids, clinically diagnosed as Liddle's syndrome. The mutation of the ENaC of this family was beta R564X. Since such case seem to be rare in the literature, detailed data are shown in this report. PMID- 11478430 TI - Cytoplasmic segment interactions in the alpha1-subunit of the rat Na+, K+-atpase. AB - The currently accepted topographical model for the organization of the alpha subunit of the Na+, K+-ATPase in the membrane considers that the protein has ten transmembrane segments and six cytoplasmic loops. Evidence of interaction between the cytoplasmic regions may contribute to a better understanding of the structure/function relationship of this protein. In this study, the first four cytoplasmic segments (C1, C2, C3 and C4) of the rat alpha1 subunit were expressed in Escherichia Coli. The large cytoplasmic loop between transmembrane segments four and five (C3) retained its native structure as demonstrated by the ability of ATP to protect against chemical modification by Fluorescein 5-isothiocyanate (FITC). Interaction studies were conducted by an overlay assay (Far Western blots) and surface plasmon resonance technology. We observed that C3 interacts with the N-terminal segment of the Na+, K+-ATPase, C1; and that both C1 and C3 interact with the cytoplasmic segments C2 and C4. PMID- 11478431 TI - Low-density lipoprotein particle size, triglyceride-rich lipoproteins, and glucose tolerance in non-diabetic men with essential hypertension. AB - The aim of the study is to investigate serum lipoproteins abnormalities including low-density lipoprotein (LDL) particle size, and their relationship with other cardiovascular risk factors in men with essential hypertension. Plasma glucose and serum insulin levels during oral glucose tolerance test (OGTT), serum lipoprotein(a), apolipoprotein (apo) A-I. apo B. cholesterol and triglycerides in serum and in lipoproteins, and LDL particle diameter were measured in thirty eight consecutive newly-diagnosed non-diabetic untreated hypertensive men and 38 healthy male controls. Plasma glucose at baseline, 60 and 120 min during OGTT was significantly higher in patients than controls whereas serum insulin levels did not differ between patients and controls. Serum apo B and triglycerides were significantly raised in patients compared with controls (1.08 +/- 0.17 g/L [mean +/- SD] vs 0.97 +/- 0.22 g/L. p < 0.05, and 1.56 +/- 0.90 mmol/L vs 1.15 +/- 0.57 mmol/L, p < 0.05, respectively). Very-low-density lipoprotein (VLDL) triglycerides and LDL-cholesterol were increased in patients compared with controls (0.89 +/- 0.79 mmol/L and 0.54 +/- 0.35 mmol/L, p < 0.05, and 4.08 +/- 0.85 mmol/L and 3.60 +/- 0.92 mmol/L, p < 0.05, respectively) whereas high density lipoprotein (HDL) cholesterol was lower in patients compared with controls 0.95 +/- 0.22 mmol/L and 1.07 +/- 0.20 mmol/L, p < 0.05). Adjustment for body mass index, abdominal/hip perimeter ratio and area under the glucose curve did not attenuate the relationship between hypertension and VLDL-triglycerides. Six patients and two controls had a mean LDL diameter < or = 25.5 nm and in the former serum triglycerides ranged from 1.86 mmol/L to 2.37 mmol/L. Mean LDL particle diameter in both patients and controls showed an inverse relationship with log-transformed serum triglycerides (r = - 0.51, p < 0.001 and r = - 0.47, p < 0.005, respectively). Among patients, those with serum triglycerides > or = [corrected] 1.58 mmol/L had a lesser mean LDL diameter than those with triglycerides above this threshold (25.78 +/- 0.47 nm vs 26.30 +/- 0.35 nm, p < 0.001). Higher plasma glucose, serum apo B and LDL-cholesterol as well as the decrease in serum HDL-cholesterol in patients with hypertension are consistent with high coronary heart disease risk. Not only mild hypertriglyceridemia but also high-normal serum triglycerides in themselves or as a surrogate of a predominance of small dense LDL particles in plasma convey an additional risk for cardiovascular disease in hypertensive patients even though routine plasma lipids are within or near normal range. PMID- 11478432 TI - Structural changes in the kidney induced by coarctation hypertension. AB - We investigated structural alterations in renal tissue identifying the morphological and histological changes in the non-ischemic kidney (NIK) and their potential significance in aortic coarctation-hypertensive rats (HR). HR's mean arterial pressure (MAP) was higher compared with sham operated rats (SR). An oral 10 mg/kg/day losartan (LOS) dose diminished but not reverted MAP. Hypertrophy was noted in HR NIK's with significant weight increase (p<0.01). The ratio IK/NIK in HR's decreased 22% (p<0.01). LOS proved to cause no ischemic kidney (IK) modification nor did it revert NIK hypertrophy. NIK in HR's presented glomerulosclerosis, mesangial proliferation and arteriolar thickening reverted by LOS. The stereological study of afferent NIK arterioles showed hypertrophy and an increase in the wall/lumen ratio without lumen modification. LOS diminished wall thickness. LOS-induced decrease of NIK alterations might result from arteriosclerosis regression, the media/lumen ratio. glomerulosclerosis and mesangial proliferation dependent on angiotensin 11. PMID- 11478433 TI - Evaluation of laboratory techniques for differentiation between Entamoeba histolytica and Entamoeba dispar. AB - Incorporation of clinical, epidemiological and serological parameters to the studies of Entamoeba histolytica resulted in the conclusion that species complex comprising two morphologically identical amoeba were implicated with the disease. The two organisms are E. histolytica and E. dispar. The former is a pathogen and responsible for invasive amoebiasis while the later is a gut commensal. Clearly accurate diagnostic tools are required to distinguish the two species. The use of a stool ELISA has been shown to be useful. A monoclonal enzyme linked immunosorbent assay based kit to detect antigen of E.histolytica only in stool was evaluated in comparison to the results of the microscopical examination of the stools and that of enzyme linked immunosorbent assay used to detect the anti- E. histolytica IgG in serum. This study demonstrated that E. dispar is prevalent in the community and offers promise for E. histolytica monoclonal enzyme immunoassay for the qualitative and semiquantitative determination of sensu lato antigen in stool as a sensitive tool for detection and distinction of E. histolytica from E. dispar infections. PMID- 11478434 TI - Clinical features, tissue smear and competitive enzyme linked immunosorbent assay as dignostic and prognostic procedures for visceral leishmaniasis in Yemen. AB - Twenty six Yemeni patients with clinically visceral leishmaniasis (VL) and laboratory abnormalities were subjected to L. donovani species-specific monoclonal antibody (D2) and a competitive enzyme-linked immunosorbent assay (C ELISA). The results were compared with those obtained by microscopy Giemsa stained tissue smears of bone marrow or splenic aspiration. Of the patients, 51 (82.3%) were positive and 3 (4.8%) were negative by both microscopy and C-ELISA. An additional 8 (12.9%) who were negative by microscopy were positive by C-ELISA. 59 patients were treated with sodium antimony gluconate, 45 (76.3%) were drug responsive and 14 (23.7%) were drug resistant. The C-ELISA of 17 patients before and after chemotherapy showed a significant decrease in percent inhibition of monoclonal antibodies D2 in drug responsive patients. In drug-unresponsive patients the percent inhibition of D2 was unchanged or slightly increased. The results indicate that C- ELISA is more sensitive and specific than microscopy, especially for early diagnosis of VL and to evaluate the success of drug treatment. PMID- 11478435 TI - Prevalence of toxoplasma infection in Qualyobia governorate, Egypt. AB - ELISA for toxoplasmosis was done on 152 randomly selected individuals, 31 full term parturient and 38 aborted or prematurely delivered women. Seropositivity to specific antitoxoplasma IgG antibodies was observed in 57.9%, 58.1% and 44.7% of random, full term and aborted samples respectively. Only 10.5%, 6.5% and 23.7% were found seropositive for specific antitoxoplasma IgM, respectively. The risk of feto-maternal transmission was very high (50%). PMID- 11478436 TI - Investigation of blood parasites in livestock infested with argasid and ixodid ticks in Egypt. AB - Nineteen species and subspecies of ixodid and argasid ticks, Hyalomma dromedarii, H. impeltatum, H. anatolicum excavatum, H. a. anatolicum, H. truncatum, H. m. marginatum, H. m. rufipes, H. m. turanicum, Boophilus annulatus, Rhipicephalus sanguineus, R. turanicus, R. guilhoni, R. camicasi, Amblyomma lipidum, A. marmoreum, A. vareigatum, Argas persicus, A. hermanii and A. arboreus were collected from different localities in five governorates, Giza, Sharkia, Ismailia, El Beheira and Sinai. Hyalomma species were found on camels (98.6%) and cows (1.4%). B. annulatus was found only on cows (100%), Rhipicephalus species on dogs (89.8%), camels (8.2%) and sheep (2.0%), Amblyomma species were found on imported camels (100%), and Argas species on chickens (70.6%), herons (18.8%) and pigeons (10.6%). Examination of camels, cows, sheep, and chickens infested with ticks showed Theileria annulata (rod and ovoid), Babesia bigemmina (ring), B. ovis (ovoid), and Babesiosoma gallinarum (dot-like), respectively. Haemoproteus columbae were found in pigeons. These parasites were determined by microscope imaged. PMID- 11478437 TI - Ivermectin and clorsulon (ivomec super) in treatment of goats naturally infested with scab mites and biting lice. AB - Five goats naturally infested with Sarcoptes scabiei, Psoroptes ovis and Bovicola caprae were treated with a combination of ivermectin and clorsulon. A subcutaneous injection in a dose of 1 ml/50 kg live weight was given. This provides dose levels of 200 ug ivermectin and 2 mg clorsulon. Four goats were cured from the three ectoparasites within one to two weeks. The last elder goat, which was anaemic and naturally infected with Trichostrongylus colubriformis, was still infested with the two scab mites, but was free from B. caprae. A postal dose of 0.5 ml/50 kg live weight was given one week later (=3 weeks after the 1st injection). This goat was cured within a week, but still passed some T. colibriformis eggs. The result was discussed. PMID- 11478438 TI - Immunohistochemical study of schistosomal soluble egg antigen in male genital organs of Schistosomiasis mansoni infected mice. AB - Seventy Swiss outbred mice were used for demonstration of circulating soluble egg antigen (CSEA) in serum and testes of S. mansoni infected mice and to study the possible role of this antigen (CSEA) in the pathogenesis of reproductive organ lesions using two recently described monoclonal antibodies to repetitive carbohydrate epitopes of S. mansoni SEA (114-5B-1A and 114-4D12-A). Ten mice as a control group and the other 60 mice were infected with 100 S. mansoni cercariae per mouse by subcutaneous injection. One group was sacrificed in each of 6,8,10,12,14 and 16 weeks post-infection (P.I.) while control mice were killed at one time. The sacrificed mice were bled and their sera were used for detection of CSEA by ELISA. The testes were dissected and paraffin blocks for histopathological study and indirect immunofluorescence technique. The mean level of CSEA in the serum of infected mice started to appear 6 weeks P.I. and gradually increased till 16 weeks P.I. This was coincided with the appearance of the fluorescent deposits in blood vessels of testis at 8 week P.I. and later in the stroma of testis from the 10th week onwards. These data together with histopathological findings with absence of egg in all examined testicular sections emphasize the possible role of CSEA in pathogenesis of such lesions. PMID- 11478439 TI - Dynamic of cercarial production of Schistosoma mansoni in Biomphalaria alexandrina. AB - The susceptibility of B. alexandrina to infection with S. mansoni miracidia varies due to biological & ecological parameters. In case of snail infection in less than 4 or 5 months productivity doubled between 1 and 2 miracidium, but with more than 2 miracidia with practically no increase in productivity. In more than 5 months, there no was discernible difference in productivity between monomiracidial and polymiracidial infections. The production of cercariae was characterized by very irregular daily emissions and by the alternation of periods of high and low emissions, which gave rise to a production rhythm. PMID- 11478440 TI - Ecto- and blood parasites affecting Meriones rex trapped in Najran, Saudi Arabia. AB - Meriones rex or king jird is endemic to Arabia, confined to the south-west of the peninsula. Examination of 25 jirds for arthropod-ectoparasites showed Xenopsylla astia, Ctenocephalides arabicus, Ornithonyssus bacoti and tick nymphs with indices of 0.6, 1.6, 0.64 and 0.24 respectively. The ectoparasites were more on female jirds than on males with indices of 3.8 and 2.0 respectively. Indirect haemagglutination tests for anti-Toxoplsama and anti-Leishmania antibodies showed positive reactions in 5 (20.0%) and 2 (8.0%) jirds respectively. However, neither skin lesion nor protozoal parasites were detected in tissue smears of liver and spleen. The ectoparasites encountered and the sero-positive reactions to Toxoplasma and Leishmania denote that the king jird, Meriones rex is of some medical and veterinary importance. PMID- 11478441 TI - Experimental amoebic liver abcess produced by oral administration of Entamoeba histolytica cysts. AB - To determine the possibility of amoebic invasion and liver-abscess formation Swiss albino mice were infected orally with E. histolytica cysts isolated from human stools. Parasitological and histopathological changes in mice colon and liver tissues were sequentially followed. Three weeks postinfection (p.i) 5% of immunocompetent and all cortisonized immunosuppressed mice passed the parasite in their stools. Only 70% of the latter group of mice sacrificed at that time developed invasive intestinal amoebiasis. At the end of the experiment (12 weeks p.i.) 100% of the remaining immunosuppressed animals developed the same intestinal pathology. Amoebic liver abscess was detected in 62.5% of them. Oral inoculation of E. histolytica cysts constitutes an easy highly reproducible procedure for inducing liver abscess in immunosuppressed mice. PMID- 11478442 TI - Effect of a broad spectrum antiparasitic drug "ivermectin" in acute and chronic experimental giardiasis using different dose regimens. AB - In this study, the maximal excretion of Giardia lamblia cysts was three weeks post infection (p.i). Administration of ivermectin subcutaneously proved to be effective in treating hamsters infected for 2 and 3 weeks respectively. It was found that the dose of 300 microg/kg b.w. was much more efficient than 200 mg/kg b.w. The cure rate was 99.1% in the former, and 98.7% in the later. The difference was statistically significant. In chronic giardiasis where infection was kept for 6 weeks, the cure rate was 99.5% two weeks after treatment with 300 g/kg b.w. of ivermectin. Assessment of cure was performed also by histopathological examination of upper 2/3 of small intestine of the hamsters. Localization and counting of the parasite were carried out immunohistochemically. The mean number of trophozoites decreased markedly after treatment with the large dose either acute or chronic giardiasis. PMID- 11478443 TI - Two species of tabanids (order: Diptera) in Aswan District, Egypt. AB - Tabanus taeniola and Haematopota minuscula were trapped on camels and equines at daytime during summer of 2000. Many species of Tabanus and few species of Haematopota were reported before in Egypt. This paper stresses on the medical and veterinary importance of these vectors. PMID- 11478444 TI - Effect of double infection with Schistosoma mansoni and Echinostoma liei on some physiological parameters of Biomphalaria alexandrina. AB - The survival rate and fecundity of B. alexandrina were greatly influenced when exposed to either S. mansoni or E. liei miracidia. The snails exhibited much lower survival rate and fecundity when double exposed to both S. mansoni and E. liei miracidia than single exposure and control snails. The results indicated a disruption in the snail metabolism due to exposure to S. mansoni and E. liei miracidia and this effect was more pronounced in case of double exposure to the two parasites. Protein concentrations in hemolymph and tissues significantly reduced in all exposed snail groups than in the control group. A significant elevation in the levels of aspartate aminotransferase (ASAT) and alanine aminotransferase (Asat) enzymes was recorded in hemolymph and tissues of exposed snail than unexposed (control snails). The ASAT/ALAT ratios in tissue and hemolymph of single-exposed and unexposed B. alexandrina did not exceed 1, while it increased up to 1.18 in hemolymph of double exposed snails. There were significant increases in the levels of acid and alkaline phosphatases enzymes in exposed snails. PMID- 11478445 TI - First record of actinosporean stages of fish myxosporean parasites (Myxozoa: Myxosporea) in Egypt. AB - In the present study the actinosporean infestation of the oligochaete fauna has been studied. For first time in Egypt, a total of 6 different types of actinospores were isolated from the filtrated water contained oligochaetes Limnodrilus hoffmeisteri, Tubifex tubifex, Branchiura sowerbyei and Dero digitata. These new types are classified to hexactinospores and triactinospores. The drawings depicting these actinospore types are constructed and their characteristic dimensions are calculated. The obtained actinospore types are not identical with those described neither in Europe nor in Japan. Throughout the present study it could not decide which actinosporean represent any of species of myxosporean parasites in the given area which required a series of experiments to develop a model for each myxospore-actinospore life cycle. PMID- 11478446 TI - Studies on chicken acquired resistance to Argas (persicargas) persicus Latereille (Acari: Argasidae) due to repeated infestation. AB - Spring chickens were used for feeding Argas persicus (females) daily over one week during both winter and summer seasons. Acquired resistance to ticks was monitored by: 1) failure of ticks to replenish a blood meal from chickens bitten repeatedly by the infesting ticks during winter and summer seasons; 2) measurements of anti-tick activity in the chicken sera; 3) detection of changes in their serum proteins. Chickens were bled after the 4th feeding, during the 1st, 2nd, 3rd and 4th weeks post-feeding. The titre of anti-tick antibody was determined in the chicken sera by an enzyme-linked immunosorbent assay (ELISA) technique. The change in sera protein bands after Argas persicus female repeated feeding was studied by the use of 10% SDS polyacrylamide gel electrophoresis. The results showed that the nonfeeding percentage in A. persicus was significant in both winter and summer seasons. The highest concentration of antibodies against A. persicus was detected after the fourth feeding and the lowest titre was reported in sera collected after the fourth week in both seasons. Infested chicken serum proteins electrophoresis showed different patterns of separation from the non-infested chickens. The protein bands of the noninfested chicken sera had 5 and 10 bands in the winter and summer seasons, but in infested chicken sera, it ranged between 12-17 and 14-18 bands in winter and summer seasons respectively. PMID- 11478447 TI - Bionomics of ticks collected from Sinai Peninsula: 2- abundance, attachment sites, and density estimators of ticks infesting Arabian camels. AB - Ten camels were examined monthly from January to December 1999, in Beer Lehfen North Sinai Gavernorate. A total of 5468 adult ticks of 6 species and subspecies of genus Hyalomma; H. dromedrii, H. ipmeltatum, H. a. excavatum, H. a. anatolicum, H. schulzei, and H. marginatum rufipes, in addition to 115 Hyalomma nymphs' species were collected. Of these, the first 4 types represented about 96.2% of the total collection. The infestation rate was higher during the period from March to November. The average monthly infestation rate was 22.78+/-3. The large number of the adult ticks were concentrated on tail, brisket, anus, and udder which totaled 68.2%. Attachment sites of males and females were not significantly correlated in most species. Nymphs were localized in few sites, whereas 91.6% of the total nymphs were attached to hump, neck, head (ear), and side. H. dromedarii, H. impeltatum, and H. a. excavatum were found on most of the body sites, tail, brisket, udder, and anus, totaling 79.9%. Males exceeded females in all sites, and were alone on neck and legs. Females were the only stage collected from udder. Both sex of H. a. anatolicum were collected from tail. H. schulzei and H. m. rufipes were concentrated in brisket, udder, anus, and tail. PMID- 11478448 TI - Identification of factors and groups at risk of infection with Schistosoma mansoni in urban Sharkia Governorate. AB - The main objective is to determine the relationship between biological, socioeconomic and behavioral factors and the prevalence of Schistosoma mansoni in urban areas. Sharkia Governorate was stratified according to the population size into large and small sized cities. A questionnaire was used to collect information on each individual included in the study and specimens of stool were also collected and examined parasitologically for the detection of S. mansoni eggs. Those found infected were treated with praziquantal. The overall prevalence was 6.8% ranging from 2.8% in large sized cities to 10.3% in small sized ones. The geometric mean egg count was almost similar in both types of cities (45.8 eggs per gm stool). Univariate and multivariate analysis identified several variables that were associated with the occurrence of S. mansoni infection in both large and small sized cities. groups at risk of being infected were identified. The data of the present study are valuable in finding some of the risk factors responsible for the occurrence of schistosomiasis in urban areas as well as for finding the high risk groups, all of which are important for planning the needed control strategies. PMID- 11478449 TI - Study of soluble adhesion molecules in some intestinal and tissue helminthes. AB - Soluble adhesion molecules namely soluble intercellular adhesion molecule-1 (sICAM-1) and soluble E-selectin (sELAM-1) were assayed in hydatid patients with or without complications. It was found that sICAM-1 was significantly increased in patients with hydatid cysts as compared to the control group. In lymphoedemic filariasis cases both sICAM-1 and sELAM-1 showed highly significant increase more than the control group. However, non of both soluble adhesion molecules were significantly elevated in patients with ascariasis, hymenolepiasis, heterophyiasis and strongyloidiasis as compared to controls. The results indicated that SICAM-1 & S ELAM-1 are useful markers for hydatidosis and filariasis, but not for ascariasis, hymenolepiasis, heterophyiasis and strongyloidiasis PMID- 11478450 TI - Studies on lice infesting goats in North Sinai. AB - In North Sinai, in a total of 304 goats, 218 (71.7%) were infested with 3 species of lice. These were Bovicola caprae (31.6%), Linognathus africanus (24.01%) and L. stenopsis (10.53%). Infestation with more than one species of lice was also seen. The highly infested season was winter (56.9%) and the least was spring (6.9%). The highly infested body site was hip (17.7%) and the least infested one was britch (6.6%). The economic loss of goats infested with lice was discussed. PMID- 11478451 TI - Biochemical changes of Culex pipiens larvae treated with oil and oleo-resin extracts of Myrrh Commiphora molmol. AB - The mosquitocidal plant extracts of the Myrrh, Commiphora molmol namely; oil and oleo-resin, were proved to demonstrate larvicidal activity against Culex pipiens larvae. Oleo-resin was found to induce significant higher toxic action than oil. Solvent (cremophore EL) did not perform any toxic activity. The Impact of both oleo-resin and oil extracts on the protein profile of the treated larvae was evaluated in order to explain their mode of action. Electrophoretic analysis of total proteins, lipoproteins and glycoproteins revealed inhibitory action of the used plant extracts on the protein contents. Larvicidal activity of the oleo resin and oil was explained as to be related to the loss of certain enzymes inhibited by these extracts which affect the metabolic processes. PMID- 11478452 TI - Application of Bacillus thuringiensis in poultry houses as a biological control agent against the housefly, Musca domestica sorbens. AB - The efficiency of Bacillus thuringiensis (B.t) as a microbial housefly control agent was evaluated in one of the natural maggot breeding medium, namely poultry faeces. Two commercial formulations, a wettable powder and a liquid concentrate were assayed against Musca domestica sorbens in synthetic fly breeding madia and in chicken faeces. The latter was found to favour maggot breeding when compared with the synthetic one. Both formulations resulted in significant reduction in maggot numbers,, pupal development and adult emergence. Concentration was found significantly affecting adult emergence. The liquid concentrate induced higher larvicidal activity than dry powder formulation. Contaminated faeces produced from orally fed chickens were found significantly toxic to the breeding maggots, reaching maximum larvicidal activity till the 4th day post-feeding faeces, and continued in less activity till the 6th day. Adding B.t. in chicken food in poultry mass breeding is recommended for housefly control. PMID- 11478453 TI - A study on Trichomoniasis vaginalis and female infertility. AB - A total of 280 patients, 240 infertile and 40 pregnant were subjected to thorough history taking, general and local examination for exclusion of organic lesion, laboratory investigations to exclude parasitic, bacterial and fungal infections. Sterile vaginal swab from the posterior fornix was taken, and examined by wet smear preparation, Giemsa staining and cultivation on C.P.L.M. medium for trichomoniasis infection. The mean age of the infertile group was 25.75+/-3.92, and of the control group was 21.6+/-2.38 (in years). The mean duration of infertility was 2.81+/-1.51 (years). Out of 240 infertile women, 18.75% complained of discharge, 17.5% itching, 15.42% dysuria, 14.58% dyspareunia, and 10% had cervical lesion. Of the 40 controls, 5% complained of discharge, 2.5% complained of itching, dysuria, dyspareunia, but none had cervical lesion. Of the total cases (280), 36 (12.9%) had T. vaginalis. The clinical data observed were significantly higher among the infertile group than the control group. Cultures were positive in 14.58% of the infertile group and 2.5% in the control group. The difference between the 2 groups was statistically significant. No doubt, T. vaginalis plays an important role in female infertility. PMID- 11478454 TI - A comparative study between laparoscopic versus open appendicectomy in men. AB - One hundred men between the age of 15 and 65 admitted to Ain Shams Specialized Hospital and El Demerdash Hospital, Cairo, and to the Saudi German Hospital, Saudi Arabia between November 1998 and December 2000 were randomized to undergo either open or laparoscopic appendicectomy. Both groups were compared in terms of clinical parameters, duration of anaesthesia, operative time, duration of ileus and length of hospital stay. The histologic confirmation of appendicitis was present in 92% in both groups. Laparoscopic appendectomy required significantly longer anaesthetic time (78 minutes versus 51) and operating time (49 minutes versus 23) compared with open appendectomy. No significant difference was noticed between laparoscopic and open appendectomy groups in the recovery of bowel function (24 hours versus 21) and in the length of hospital stay (4.9 days versus 5.3). The result showed no significant advantages of laparoscopic appendectomy over open appendectomy for the treatment of male patients with suspected appendicitis. PMID- 11478455 TI - Observations on the structure and histology of the female genital system of the cattle tick Boophilus annulatus (Say). AB - Female genital system of Boophilus annulatus was examined grossly on the day of drop-off from the host and grossly and microscopically on 4th day. It consists of an ovary, 2 oviducts, a common oviduct, a connecting tube, a seminal receptacle, a vagina divided into cervical and vestibular parts, 2 accessory glands and Gene's organ. The ovary oocytes undergo excessive growth during the 4 days after drop-off from the host. Oocytes accumulate eosinophilic material during growth and undergo histological changes similar to those seen in other ixodid ticks. Further ultrastructural studies are required to understand the vector capacity of this species in transmitting disease agents. PMID- 11478456 TI - Circulating filarial antigens for monitoring the efficacy of ivermectin in treatment of filariasis. AB - Seventy-three asymptomatic bancroftian filariasis patients with positive microfilaria in their blood films were included. The patients were randomly divided into 2 groups: ivermectin group (50 cases) given 2 doses each of 100 ug/kg body weight, 3 months apart, and 23 cases had 2 doses of placebo. The study was run blindly for one year. The initial mean microfilaria (MF) count was 111/ml. At 3 months after ivermectin therapy, mean MF became 7.8/ml and 24% of ivermectin treated cases had no detectable MF (P <0.05). At 6, 9 and 12 months, the mean MF count became 4.1, 6.5 and 11/ml with amicrofilaria in 54%, 42% and 40% of treated cases respectively (P <0.05). On the other hand, no statistically significant change in the mean MF count in placebo group was detected. The routine laboratory investigations were unchanged or slightly improved at 3 and 6 months. Side effects after the first dose of ivermectin were mild fever in 16% and weakness in 20%. None was recorded after the second dose. Circulating filarial antigens could be detected in 66% of cases before treatment, as all cases with high microfilaremia had positive antigenemia. The mean antigen level started to decline significantly after 9 months post treatment. At the end of the study (one-year), all negative microfilaremic cases had negative antigen levels, indicating that detection of antigen in-patients sera is a very good indicator of cure and efficacy of the drug. PMID- 11478457 TI - Studies on patients with Schistosomiasis mansoni, HCV and/or typhoid fever. AB - Two groups of patients were studied. First one included 50 schistosomiasis mansoni patients, 30 with simple infection, 10 with splenomegaly and with ascites. Second group included 111 patients of whom 20 with pure S. mansoni, 27 with pure HCV infection, 54 with mixed infection of schistosomiasis and HCV and 10 with schistosomiasis, HCV and typhoid fever. Serum transaminases and anti-HCV antibodies performed, showed anti-HCV raised levels in 10% of simple schistosomiasis, 60% in splenomegalic patients, 80% in ascites patients, and 7.1% in controls. Liver function tests in first group were within normal range except in those with ascites. In second group, liver function tests was norma in pure schistosomiasis patients, in pure HCV patients serum bilirubin was normal in 22.2%, AST, ALT and alkaline phosphatase were higher. In mixed infection, serum bilirubin was normal in 18.5%, serum transaminases were higher and alkaline phosphatase was normal among 77.7%. Patients with typhoid fever, HCV and schistosomiasis (12.6%) showed significant increase of liver function as compared with each of pure HCV or HCV and schistosomasis. Results were discussed. PMID- 11478458 TI - Effect of ammonium chloride on Biomphalaria alexandrina and on its infection with Schistosoma mansoni and Echinostoma liei miracidia. AB - Ammonium chloride (NH4Cl) has molluscicidal activity against B. alexandrina. The LC50 and LC90 recorded of this salt were found to be 90 ppm and 130 ppm, respectively. Maintaining of B. alexandrina at low concentrations of NH4Cl (5, 10, 15 and 20 ppm) greatly reduced their survival rate and fecundity. The net reproductive rate (Ro) [sigmaIx Mx] was deleteriously affected. This rate was significantly reduced than that of control snails in all tested snail groups. The reduction in Ro was 86.9%, 90.8%, 93.9% and 96.9%, respectively. The susceptibility of B. alexandrina to infection with S. mansoni and E. liei miracidia and infectivity of these two parasites were greatly reduced. Increasing the salt concentration increased this reduction. The magnitude of reduction in infection rate was lower in case of E. liei than that in S. mansoni indicating that E. liei is more tolerant to the effect of this salt than S. mansoni. PMID- 11478459 TI - Effect of ozone on the viability of some protozoa in drinking water. AB - Water samples from household tanks in several districts of Alexandria, were examined for protozoa contamination and study to the effect of ozone and chlorine on their viability. The parasitic protozoa encountered were Giardia (56%), Cryptosporidia (50%), Blastocystis (12%), Cyclospora (9%) and Microsporidia (3%). Distilled water was treated by ozone or chlorine. Ozonated-water samples were prepared at a concentration of 1 ppm after contact times with ozone of three, five, seven and nine minutes. Chlorine concentrations were 4 and 8 ppm. Parasites were incubated overnight at room temperature with the treated distilled water. Viability of the parasites was assessed by examining intestinal sections of infected animals. The best results were obtained by using ozonated water after a contact time of nine minutes. Giardia, Cryptosporidia and Microsporidia were completely inactivated, while infectivity of Cyclospora and Blastocystis was markedly reduced. It is suggested that ozone at an appropriate concentration inactivate pathogenic protozoa in water. PMID- 11478460 TI - Glycerol Jelly (GJ) mount: a new and simple method for routine stool examination using a modified glycerol jelly reagent. AB - Wet mount is the basic primary technique for stool examination in laboratories, allowing only the use of X10 and X40 objectives which do not sometimes reveal relevant details to make an exact identification of certain protozoa. In a modification of the Glycerol Jelly (GJ) reagent, which is used in permanent preparation of helminths, five concentrations were prepared and tested for fixing the cover glass instantly while maintaining the high translucency of the fecal films. GJ reagent (7 gm gelatine dissolved in 50 ml boiling water added to 10 ml glycerol) gave satisfactory results especially with iodine and alkaline Methylene blue mounts which stained the cytological structures of protozoa while the GJ reagent enabled the examiner to use X100 oil immersion objective immediately and consequently identify protozoa with certainty and make an accurate identification. Identification of Cryptosporidium parvum oocysts by GJ wet mount, inspite of its small size, was the most impressive. GJ fecal films were examined up to 8 weeks of preparation and they were valid and reliable. GJ mount is an easy, fast and cheap technique for examining the fecal direct smear with the oil immersion lense. PMID- 11478461 TI - Morphology, histochemistry and infectivity of Blastocystis hominis cyst. AB - Different morphological forms of Blastocystis hominis had been identified in human stool samples. These included both cystic and trophic stages. The latter was induced to encyst by keeping them in potassium dichromate solution for two weeks. Suspected of being the infective stage, cysts were studied in more detail as regards their morphology using both light and electron microscopy. Histochemistry and infectivity studies were also carried out. Light microscopy revealed the cysts to be ovoid or round 5-7 microm with thick cyst wall and a single nucleus. The induced cysts were morphologically similar to the cysts present in the stool. By electron microscope, the cyst wall was evident surrounded by an additional fibrillar coat. The cytoplasm contained one nucleus, many mitochondria, glycogen deposits and a number of variable sized vacuoles. Histochemical studies detected carbohydrates in the cyst wall and fat globules in the cytoplasm. Oral inoculation of albino mice with these cysts led to inflammatory changes in the large and small intestine. The parasite was found at the mucosal epithelium but with no invasion. Different forms of the parasite were detected in the lumen of the intestine. PMID- 11478462 TI - Toxoplasma antibodies in normal and complicated pregnancy. AB - A total of 70 women with complicated pregnancy and 20 ones with normal pregnancy were selected. They were all RH positive and free from syphilis, brucellosis and malignany. Both groups were subjected to detection of anti-Toxoplasma antibodies levels by the indirect haemagglutination tests (IHAT-IgG) and by ELISA (IgG and IgM). The results showed that all controls (normal pregnancy) were IHA-IgG and ELISA-IgM negative. However, two of the controls (10%) were ELISA. IgG positive. As to the women with complicated pregnancy 68/70 or 97.1% were IHAT positive. When using ELISA, 57/70 or 81.4% were IgG positive and 42/70 or 60% were IgM positive. The results revealed a significant difference between the complicated cases and controls. The diagnosis of abortion, stillbirth, premature labour, or neonatal death due to Toxoplasma infection needs careful and laborious evaluation. The use of ELISA (IgG & IgM) is a must, as it is more sensitive and specific than IHAT. PMID- 11478463 TI - Promoting our profession: we have a great story to tell. PMID- 11478464 TI - How to evaluate the safety, efficacy, and quality of functional foods and their ingredients. PMID- 11478465 TI - Getting into the media spotlight: the ADA's public relations team promotes the Association as the most recognized source for nutrition information. PMID- 11478466 TI - MNT works: marketing resources spread the message. PMID- 11478467 TI - The Dietary Guidelines for Americans: a basis for US nutrition policy. PMID- 11478468 TI - Time for dietetics and mental health alliance? PMID- 11478469 TI - The establishment of a "transition diet". PMID- 11478470 TI - The development of TPN: an interview with pioneer surgical nutritionist Jonathan E. Rhoads, MD. [Interview by Carolyn T. Spencer and Charlene Compher]. PMID- 11478471 TI - Evaluation of validity of items for a food behavior checklist. AB - OBJECTIVES: To evaluate the validity of food behavior items, using a biological measure (serum carotenoids) as the criterion for validity of fruit and vegetable intake, and the results from multiple 24-hour recalls to test convergent validity with nutrient intake. DESIGN: Participants responded to 39 food-behavior questions and later completed three 1-day dietary recalls. Serum carotenoid levels were determined for a 59% randomly selected subsample. SUBJECTS/SETTING: A convenience sample of 100 English-speaking, low-income women participating in a Food Stamp Nutrition Education Program in 8 California counties. Statistical analyses Spearman correlation coefficients were calculated between responses to the food behavior items and (a) serum carotenoid levels and (b) mean nutrient intakes from the 24-hour recalls. Cronbach's coefficient alpha was determined for items within broad food behavior topics. RESULTS: Responses to 10 food behavior items were significantly correlated with serum carotenoid levels (correlations greater than 0.45 were found for choosing low-fat foods and a self-evaluation of overall dietary quality). An additional 12 items showed hypothesized associations with the 24-hour recall data (with a maximum correlation 0.50 for number of eggs per week and dietary cholesterol). Cronbach's coefficient alpha ranged from 0.28 (for 5 fat and cholesterol items) to 0.79 (for 9 fruit and vegetable items). APPLICATIONS/CONCLUSIONS: Nutrition professionals can use these methods to validate items for food behavior checklists for specific populations. The items described here may be useful when designing instruments to administer to low income women in a community setting. PMID- 11478472 TI - Predictors of self-initiated, healthful dietary change. AB - OBJECTIVE: To examine demographic and psychosocial factors that predict healthful dietary change. DESIGN: A cohort study, examining how factors assessed at baseline predicted change in fat-related dietary habits and fruit and vegetable intakes 2 years later. PARTICIPANTS: Participants were recruited in 1995 and 1996 by random-digit dialing (response rate 0.63), and followed-up in 1997 and 1998 (follow-up rate 0.82). The final sample included 336 men and 502 women. MAIN OUTCOME MEASURES: Fruit and vegetable intake and fat-related dietary patterns, measured by telephone-administered surveys. STATISTICAL ANALYSES: Chi2 tests and linear regression were used to test associations of baseline characteristics with dietary change. RESULTS: Fat intake (energy from fat) decreased by approximately 2 percentage points and fruits and vegetables intake increased by 0.17 servings per day (both P<.001). Changes were significantly larger among women and persons who were well educated. Persons in the maintenance stage of change and persons who believed there was a strong relationship between diet and cancer made the largest dietary changes. Use of food labels was strongly associated with fat reduction, but not with increases in fruits and vegetables. APPLICATIONS: These results suggest that food labels are useful for helping people reduce fat intake, that interventions should target persons at all stages of dietary change, and that new efforts are needed to reach men and persons who are less well educated. PMID- 11478473 TI - The effects of equal-energy portions of different breads on blood glucose levels, feelings of fullness and subsequent food intake. AB - OBJECTIVE: This study compared the effects of equal-energy portions of 7 different breads on feelings on fullness and subsequent ad libitum food intake. A satiety index score (SI) was calculated for each of the breads. DESIGN: Within subject, repeated-measures design. SUBJECTS AND INTERVENTION: Ten healthy subjects participated in the study. Subjects fasted for > or =10 hours overnight and then reported to the research center the next morning, where they first completed baseline satiety ratings, gave a fasting blood sample, and then consumed a test bread. Additional finger prick blood samples and satiety ratings were collected at 15-minute intervals over 120 minutes, after which the subjects' ad libitum intake of food was recorded. A satiety index (SI) score was calculated for each test food by dividing the area under the 120-m satiety response curve (AUC) for the test bread by the satiety AUC for the reference bread (regular white bread) and multiplying by 100%. RESULTS: The mean SI scores for the breads ranged from 100% to 561%, with regular white bread having the lowest SI score. Mean SI scores were negatively correlated with energy intake at a test meal after 120 m (r=-0.88, P<.01, n=7) and total day energy intakes (r=-0.72, P<.05, n=7). The strongest predictor of the breads' SI scores was their portion size and thus energy density. The breads' glycemic responses were not significantly associated with fullness responses. APPLICATIONS: Ingredients and food processing methods are available for the production of palatable, high-satiety versions of processed foods, which may be useful for weight control diets. SI scores would be a useful addition to food labels to indicate which foods are less likely to be overeaten and could be used by dietitians to develop weight control plans to help reduce energy intakes without increased hunger. PMID- 11478474 TI - A 7-item versus 31-item food frequency questionnaire for measuring fruit, juice, and vegetable intake among a predominantly African-American population. AB - OBJECTIVE: This study sought to determine which of 2 fruit and vegetable food frequency questionnaires (FFQs) most closely approximated intake measured by the average of four 24-hour dietary recalls. DESIGN: Participants completed either a 31-item FFQ (n = 70) or 7-item FFQ (n = 76) on 2 occasions approximately 2 weeks apart. During the interval between FFQs1 participants provided four 24-hour dietary recalls via telephone interview. SUBJECTS/SETTING: Participants were 146 persons with food preparation responsibilities in families of students in grades 3 through 5. Respondents were predominantly African-American women in Atlanta, Ga. STATISTICAL ANALYSIS: Pearson correlation coefficients of log-transformed values estimated the reliability of each FFQ and compared FFQ estimates to reference values. The intraclass correlation coefficient evaluated consistency across 24-hour recalls. RESULTS: The first FFQs overestimated intake approximately twofold. The 31-item FFQ estimates exceeded 7-item FFQ estimates by approximately 30% . Correlations with recall estimates were high for the 7-item FFQ and moderate to low for the 31-item FFQ. The second FFQ estimates were more highly correlated to reference values. From the first to the second administration, 7-item FFQ estimates dropped from 5.2 to 2.7 servings, and 31 item FFQ estimates dropped from 6.7 to 3.5 servings. Neither FFQ produced highly reliable estimates. CONCLUSIONS: Mean total fruit and vegetable consumption was closer to reference estimates for the first 7-item FFQ and the second 31-item FFQ. The 7-item FFQ correlated more highly with reference estimates than did the 31-item FFQ. Therefore, we conclude that for African-American adults, a 1-time administered FFQ using 7 broad food categories correlates more highly with reference values than a FFQ using 31 individual fruit and vegetable items. PMID- 11478475 TI - The role of whole grains in disease prevention. AB - Despite universal acceptance of the importance of whole grains in the diet, consumer knowledge of the benefits of whole grains and intake of these foods are low. This review summarizes the research supporting whole-grain consumption and gives practical suggestions about how to increase whole-grain intake. Whole-grain foods are valuable sources of nutrients that are lacking in the American diet, including dietary fiber, B vitamins, vitamin E, selenium, zinc, copper, and magnesium. Whole-grain foods also contain phytochemicals, such as phenolic compounds, that together with vitamins and minerals play important roles in disease prevention. The exact mechanisms linking whole grains to disease prevention are not known but may include gastrointestinal effects, antioxidant; protection, and intake of phytoestrogens. Dietary intake studies indicate that consumption of whole grains is far less than the recommended intake of 3 servings a day, with an average daily intake of 1 or fewer servings a day. A new whole grains health claim, allowed in July 1999 by the Food and Drug Administration, and inclusion of a whole-grain recommendation in the 2000 revision of the US Dietary Guidelines for Americans, should help increase whole-grain consumption. PMID- 11478476 TI - Entomophagy: what is it and why are people doing it? PMID- 11478477 TI - Nutritional status classification in the Department of Veterans Affairs. AB - The Department of Veterans Affairs (VA) Nutrition Status Classification scheme uses clinical data that are routinely collected on admission or shortly thereafter for quick inpatient nutrition screening. In this scheme, patients are assigned to 1 of 4 classification levels according to 7 individual indicators. The indicators include nutrition history, unintentional weight loss as a percent of usual body weight, percent of ideal body weight, diet, diagnosis, albumin, and total lymphocyte count. After ratings (1 to 4) are assigned to each of the 7 indicators, overall nutritional status for each patient is determined by an algorithm. The VA classification system includes many of the same criteria used in other nutritional status classifications. Where it differs is in the greater emphasis on the use of objective criteria and in the rigorous evaluation of reliability and validity that went into its development. Because of these extra measures, the VA classification can be used for prioritizing workload, as well as for determining staff requirements and for comparing workload and productivity across health care facilities. So that others might benefit from using this system, this article provides information on how the classification scheme was developed and explains how it is used. PMID- 11478478 TI - Communicating dietary guidelines for Americans: room for improvement. AB - This qualitative study examines consumers' attitudes about key messages, alternative messages, and various formats of the 1995 US Dietary Guidelines for Americans. Five focus groups were conducted using a discussion guide based on previous research and the research concerns of the 1995 Dietary Guidelines Advisory Committee. The presentation orders of the guidelines, guideline alternatives, and formats were varied to prevent bias. Forty consumers (age range, 25 to 43 years) participated: 3 groups of women and 2 groups of men. Participants were not familiar with the Dietary Guidelines but had heard some messages. Consumers were confused by several guideline messages. First, the messages to "Maintain or improve your weight" and "Balance the food you eat with physical activity" did not make sense to them. Second, respondents were confused by the term "balance" in several different contexts. The participants most preferred a shortened format, followed by a 2-tiered or grouped format, with the 1995 list of Dietary Guidelines falling last. Most of the Dietary Guidelines could be better communicated to the public using shorter, clearer messages, especially for the weight and physical activity, and fat guidelines. Decreasing the number of guidelines or using a tiered/grouped format could make the Dietary Guidelines more useful to consumers. PMID- 11478479 TI - Adolescents' eating patterns influence their nutrient intakes. PMID- 11478480 TI - Dietary intake and food patterns of Zuni adolescents. PMID- 11478481 TI - Assessment of firefighters' cardiovascular disease-related knowledge and behaviors. PMID- 11478482 TI - Position of the American Dietetic Association: nutrition intervention in the treatment of anorexia nervosa, bulimia nervosa, and eating disorders not otherwise specified (EDNOS). AB - More than 5 million Americans suffer from eating disorders. Five percent of females and 1% of males have anorexia nervosa, bulimia nervosa, or binge eating disorder. It is estimated that 85% of eating disorders have their onset during the adolescent age period. Although Eating Disorders fall under the category of psychiatric diagnoses, there are a number of nutritional and medical problems and issues that require the expertise of a registered dietitian. Because of the complex biopsychosocial aspects of eating disorders, the optimal assessment and ongoing management of these conditions appears to be with an interdisciplinary team consisting of professionals from medical, nursing, nutritional, and mental health disciplines (1). Medical Nutrition Therapy provided by a registered dietitian trained in the area of eating disorders plays a significant role in the treatment and management of eating disorders. The registered dietitian, however, must understand the complexities of eating disorders such as comorbid illness, medical and psychological complications, and boundary issues. The registered dietitian needs to be aware of the specific populations at risk for eating disorders and the special considerations when dealing with these individuals. PMID- 11478483 TI - Dietetics professionals have a positive perception of professional development. AB - This study summarizes the perceptions towards the Professional Development Portfolio method of recertification and attitude towards professional development of dietetic professionals as measured in the national pilot study of the Portfolio. Portfolio compared to control respondents had greater awareness, and better understanding and knowledge of the Portfolio process after using the Portfolio guide. The perception that the Portfolio process will aid in maintaining competence did not differ between groups. The perception that self reflection aided in determining career goals and confidence to conduct a learning needs assessment differed slightly between groups. Both groups had a positive attitude towards professional development and that continuing professional education (CPE) improved competence. All perceptions and attitudes were positive. The focus of CPE in foods, business, management, education, and research did affect the ability of dietetics professionals to find appropriate CPE. Rural and unemployed respondents had more difficulty finding appropriate CPE. The findings indicate that the Portfolio guide aides the dietetics professional using the Portfolio process and that the perception towards the Portfolio process is positive. Some dietetics professionals with different CPE focus had difficulty finding appropriate CPE, and this did not differ by control and Portfolio group. More innovative methods of CPE delivery may need to be considered for these practitioners. PMID- 11478484 TI - The American Society for Parenteral and Enteral Nutrition (A.S.P.E.N.) Standards of Practice for nutrition support dietitians. PMID- 11478485 TI - Effect of gastrin and anti-gastrin antibodies on proliferation of hepatocyte cell lines. AB - Gastrin (G-17) and its precursor glycine-extended gastrin (G-17-gly) have been shown to be trophic to some gastrointestinal tumors. This in vitro study assessed the effect of G-17, G-17-gly, anti-gastrin antibodies (anti-G-17), and the CCK-B receptor antagonist PD135,158 on three hepatoma cell lines (PLC/PRF/5, HepG2 and MCA-RH7777) and an embryonic liver cell line (WRL68). The pancreatic adenocarcinoma cell line AR42J was used as a positive control. G-17 and G-17-gly caused significant proliferation of AR42J and WRL68 cell lines. G-17-gly but not G-17 induced significant proliferation of the PLC/PRF/5 cell line. Anti-G-17 and PD135,158 significantly inhibited unstimulated AR42J and WRL68 cell lines. Anti-G 17 also inhibited the proliferative effects of G-17 and G-17-gly on AR42J, WRL68, and PLC/PRF/5 cell lines, whereas PD135,158 inhibited the proliferative effect of G-17 only. G-17 and G-17-gly as well as anti-G-17 and PD135,158 had no effect on HepG2 and MCA-RH77777 cell lines. It is concluded that G-17-stimulated proliferation is mediated via the CCK-B receptor and G-17-gly via a separate, as yet uncharacterized, receptor. There may therefore be a role for gastrin in embryonic hepatocellular proliferation and perhaps also in the proliferation of some hepatocellular tumors. PMID- 11478486 TI - Differential dietary effects on colonic and small bowel neoplasia in C57BL/6J Apc Min/+ mice. AB - The effects of fiber on colon cancer are controversial. Twenty 5-week old C57BL/6J Apc Min/+ mice were fed for 60 days with a commercial mouse diet (Teklad LM-485) and eight semidefined diets containing 5-10% various fibers and 20% soybean oil. Ten additional C57BL/6J congenic litter-mates were fed each diet to assay colonic SCFA. SCFA, stool bulk, and colonic tumor incidence differed only slightly among the semidefined diets despite variations in fiber content and source. However, food consumption, caloric intake, stool bulk, and SCFA were substantially increased by the Teklad diet compared with all other groups. The Teklad diet significantly increased the number of mice with colonic tumors, average number of tumors/mouse, total tumor burden, colonic atypical hyperplasia, and small bowel tumors. Mice fed high-fat, no-fiber diets had more small bowel tumors (29.8 +/- 3.1) than mice fed diets with fiber (8.2 +/- 2.1) or with low fat and no fiber (18.1 +/- 3.4) (P < 0.05 for each group). These studies suggest that fat predisposes to and fiber protects against small bowel tumors but not colon tumors in these mice. Thus, diets high in fiber or yielding high colonic luminal SCFA may not necessarily protect against colonic cancer. Furthermore, the effects of dietary fiber in Teklad appear overshadowed by some other biologically active factors in this animal model. PMID- 11478487 TI - Human T-cell lymphotropic virus-associated primary gastric lymphoma. PMID- 11478488 TI - Expression of oncogene products HER2/Neu and Ras and fibrosis-related growth factors bFGF, TGF-beta, and PDGF in bile from biliary malignancies and inflammatory disorders. AB - The expression of several growth factors and K-ras gene mutation in bile were studied to better understand the pathogenesis and improve early diagnosis of bile duct cancers. Bile samples were collected from 12 cholangiocarcinomas (CLC), 10 ampullary cancers (APC), 3 gallbladder cancers (GBC), 7 pancreatic cancers (PNC), 9 biliary tract infection (BTI), 8 biliary stone disease (ST), and 5 normal controls (NC). The highest mean value of TGF-beta in bile was in patients with BTI; the mean levels of bFGF and PDGF were highest in CLC, and patients with APC and CLC had higher expression of HER2/Neu than other groups. In bile, a K-ras gene codon 12 mutation was found in 5 of 6 (83%) cases of CLC by the PCR-RFLP method. The results suggest overexpression of bFGF, PDGF, and HER2/Neu and the presence of K-ras mutation are important for carcinogenesis of bile duct cancers, and detection of the above abnormalities in bile is helpful for early diagnosis. PMID- 11478489 TI - Assessment of liver histology in chronic alcoholics with and without hepatitis C virus infection. AB - Patients with alcoholic liver disease have a high prevalence of hepatitis C virus (HCV) infection. The histological appearances of the liver in patients with alcoholic liver disease and HCV infection are well described. However, liver histology in individuals with dual pathology, both chronic alcohol abuse and HCV infection, is less well understood. The purpose of the present study was to examine this issue and to determine if there is any correlation between specific histological features and the serum biochemical abnormalities seen in these patients. Eighty-six chronic alcoholics, 65 with HCV infection and 21 uninfected subjects, were included in the study. All patients had history of heavy alcohol abuse (consuming 80 g or more of ethanol a day for at least 10 years). The following data were collected on each patient: demographic information (age, gender, race), the amount and duration of alcohol intake, biochemical results, and liver biopsy abnormalities including the histological activity index (HAI) score. HCV-infected alcoholics were younger (P = 0.05) and were more often African American than Caucasian (P < 0.01). Alcohol consumption was significantly greater in uninfected alcoholics compared to those with HCV infection (P < 0.05). Liver histology in subjects with HCV infection showed higher HAI scores for intralobular necrosis (P = 0.008) and periportal inflammation (P = 0.004). Features of "chronic hepatitis" and focal lymphoid aggregates were more frequent in HCV-infected alcoholics (P = 0.001 for each). By contrast, cirrhosis was present in a higher proportion of uninfected alcoholics compared to those with HCV infection (P = 0.05). Histological findings of hepatic fibrosis and total HAI score showed a significant correlation with serum albumin and platelet count in HCV-infected alcoholics. Chronic alcoholics with HCV infection have specific histological appearances that can usually help distinguish these patients from uninfected alcoholics. Correlation analysis indicates that of the various laboratory tests, serum albumin and platelet counts are the best predictors of the severity of liver damage at histology. In chronic alcoholics, the development of cirrhosis is related more to the amount of alcohol consumed than to the presence of HCV infection. PMID- 11478490 TI - Selection of HCV NS5A quasispecies during IFN therapy in patients with chronic HCV. AB - Interferon treatment of chronic hepatitis C virus infection is successful in a minority of patients. The sequence of the interferon sensitivity determining region (ISDR) of the NS5A protein may determine the outcome of therapy in patients infected with HCV genotype 1. To determine whether IFN treatment caused selection of ISDR quasispecies and whether sequences bearing the putative IFN resistance motif (HCV-J sequence) were selected, we examined amino acid changes in the ISDR in patients with HCV of different genotypes with and without therapy. We found that the ISDR sequence was highly variable and variability was greatest in patients with HCV of genotype 1. IFN treatment was found to exert a selection pressure on ISDR quasispecies, but the putative interferon-resistant variant was not enriched in patients of any genotype. Hence factors other than the sequence of the ISDR region played a role in the IFN resistance of these patients. PMID- 11478491 TI - Usefulness of a liver biopsy in the evaluation of patients with elevated ALT values and serological markers of hepatitis viral infection: an AIGO study. AB - Our study objective was to determine the current use of liver biopsy in chronic viral hepatitis and how patient management is affected by the procedure. Members of the Italian Association of Hospital Gastroenterologists were asked to complete a questionnaire for patients submitted to biopsy. Their clinical diagnosis was compared with the histological diagnosis. Of 660 patients evaluated, we selected 535 cases with viral infection. Concordance of clinical vs histologic diagnosis amounted to 84.3%, with 92.7% sensitivity and 32.4% specificity rates for diagnosing chronic hepatitis; the clinical diagnosis was correct in only 24 of 57 cases with cirrhosis. In 20 cases (3.7%) additional diagnoses were provided. The biopsy was rated avoidable in 36.8% of cases. Knowledge about grading and staging was considered of value in 59.6 and 66.4% of cases, respectively; however, when it became available, the scheduled treatment with interferon was not changed in 81.7 and 80.9% of cases. In the majority of patients with abnormal ALT and infection with HCV and/or HBV, histology documents mild/moderate inflammation with minimal fibrosis and liver biopsy neither increases the accuracy of clinical diagnosis nor affects the choice of therapy. The data from this study support the view that liver biopsies are less helpful than conventionally understood. PMID- 11478492 TI - Hepatotoxicity in patients with cirrhosis, an often unrecognized problem: lessons from a fatal case related to amoxicillin/clavulanic acid. PMID- 11478493 TI - Idiopathic adulthood ductopenia: long-term follow-up after liver transplantation. PMID- 11478494 TI - Cholelithiasic disease and associated factors in a Spanish population. AB - In order to analyze the factors associated with cholelithiasic disease, 1268 participants of a population sample were studied. On univariate analysis, 11 of the 23 variables included showed a statistically significant association (P < 0.05). Five of these variables, including obesity, triglyceride level, intake of hypolipidemic drugs, and a diet rich in cholesterol and saturated fats in women, and physical exercise in men, remained significantly associated after controlling for age. On multivariate analysis among women, a positive association was found with age (P < 0.001), obesity, and the use of hypolipidemic agents (P < 0.05) and a negative one with a diet rich in cholesterol and saturated fats (P < 0.05). Among men, the same analysis revealed there was a positive association with age (P < 0.001) and triglycerides (P < 0.05) and a negative one with physical exercise (P < 0.05). In conclusion, obesity and the use of hypolipidemic agents in women and triglycerides in men, were positively associated with cholelithiasic disease, independent of age, while negative associations included the intake of cholesterol and saturated fats in women and physical exercise in men. PMID- 11478495 TI - Cholecystokinin activation of 70-kDa S6 kinase in exocrine pancreas. AB - Cholecystokinin (CCK), a known mitogen for the exocrine pancreas, is shown to activate 70-kDa S6 kinase in isolated pancreatic acini. In this study, we examined the kinetics and cellular mechanisms of CCK-induced p70 S6 kinase activation in vivo and in vitro. Fasted mice were intraperitoneally injected with 0.01-10 microg/kg CCK analoge cerulein. Cerulein caused a concentration-dependent activation of p70 S6 kinase, with the maximal effect at 1-10 microg/kg. After 1 microg/kg cerulein administration, the kinase activity was increased at 5 min, peaked at 10 min, and subsequently decreased. Cerulein also caused a rapid and transient activation of Src. Prior administration of the tyrosine kinase inhibitor herbimycin A compeletely inhibited cerulein-induced Src activation, while the inhibition of p70 S6 kinase activity was partial. Similar results were obtained with pancreatic acinar cell line AR42J cells. These results suggest that tyrosine kinases, including Src as a possible candidate, are partly implicated in the signaling pathway of CCK-induced p70 S6 kinase activation in the exocrine pancreas. PMID- 11478496 TI - Iron, zinc, and copper concentration in serum, various organs, and hair of dogs with experimentally induced exocrine pancreatic insufficiency. AB - The concentration of iron, zinc, and copper in serum, pancreas, liver, duodenum, kidneys, myocardium, brain, and hair was studied in dogs with experimentally induced exocrine pancreatic insufficiency. Exocrine pancreatic insufficiency was performed surgically in eight healthy, 8-month-old, mongrel dogs (group I). An equal number of dogs, of the same breed and age, were used as controls (group II). One month postoperatively, the dogs in group I showed symptoms of exocrine pancreatic insufficiency, confirmed by the serum Trypsin-like immunoreactivity test, and on autopsy, by histological examination of the pancreas. At the end of the experiment (20-week duration) the dogs in both groups were sacrificed. The values of serum iron, percentage transferrin saturation, and iron concentration in pancreas, duodenum, and kidneys in group I dogs were significantly higher than those in control animals. The concentrations of zinc in serum, pancreas, and myocardium and of copper in serum, pancreas, duodenum, myocardium, and hair in group I dogs were significant lower than those in control animals. Histological examination of various organs of group I dogs revealed severe atrophy and fibrosis of the pancreas, fatty infiltration of the liver, destruction and reduction in height of the villi of the duodenal epithelium, and diffuse infiltration of the duodenal lamina propria with lymphocytes and plasmocytes. PMID- 11478497 TI - Electrogastrographic characteristics in patients of stomach cancer. AB - Using a homemade electrogastrography (EGG) system, we studied the characteristics of myoelectrical rhythm in gastric cancer (GC) patients. Based on a short-term Fourier transform, recorded slow waves could be automatically analyzed to obtain the following parameters: dominant frequency/power, percent of normal rhythm (2.4 3.7 cpm), power ratio, etc. Fifty histologically confirmed GC patients (34 men, 16 women) were enrolled before surgical intervention to measure their fasting and postprandial EGG parameters for 30 min. The cancerous parameters of GC patients were then obtained postoperatively. In addition, 46 healthy subjects were enrolled for comparison. When compared to controls, GC patients had the following characteristics: absence of postprandial increase in dominant frequency (GC: 3.04 +/- 0.47 vs 3.07 +/- 0.44 cpm, NS; controls: 3.02 +/- 0.31 vs 3.21 +/- 0.25 cpm, P < 0.001), marked power response after meal (P < 0.05), and obvious power ratio (4.58 +/- 7.38 vs 2.27 +/- 2.05, P < 0.05). Multivariate analysis indicated that advanced GC was the factor responsible for the obvious dominant power enhancement after meal (P < 0.05). Other demographic, clinical, and cancerous factors did not influence EGG parameters. We conclude that apparent arrhythmia is not encountered in GC patients, although they mainly exhibit obvious postprandial power response. Advanced GC is likely responsible for this power enhancement on EGG recording. PMID- 11478498 TI - Assessment of external anal sphincter morphology in idiopathic fecal incontinence with endocoil magnetic resonance imaging. AB - The failure of external anal sphincter repair may relate to sphincter atrophy where muscle fibers are replaced by fat, seen on MRI due to the differing signals returned by fat and muscle tissue. Manometry, electrophysiology, and MRI with an endocoil were performed on 34 fecally incontinent patients with intact sphincters on endosonography. The area of the external sphincter was measured in the midcoronal plane, and the percentage fat content calculated. Sphincter muscle area correlated strongly with squeeze pressure (P < 0.001) but not with percentage fat content. There was no relationship between percentage fat and age, weight, anal sensation, squeeze pressure, sphincter length or width, or pudendal nerve terminal motor latency. There was a trend for smaller sphincters to contain a higher percentage fat content (P = 0.059). MRI has established a relationship between function and external sphincter bulk, but not fat content, although smaller muscles may contain more fat. PMID- 11478499 TI - Human duodenal motor activity in response to acid and different nutrients. AB - Duodenal motor activity in response to intraduodenal infusion of small volumes of acid and nutrients of different chemical composition was studied in 10 healthy humans, using a water-perfused catheter incorporating 20 antropyloroduodenal sideholes. Saline and dextrose did not affect motility. Acid very rapidly (in 39 +/- 11 sec) increased the number of pressure waves (P = 0.035) and antegradely propagated pressure waves (P = 0.02). After lipid infusion a considerable lag time (163 +/- 81 sec) was observed, followed by a prominent increase in duodenal pressure waves (P = 0.02) and antegradely propagated pressure waves (P = 0.002). Furthermore, lipid-induced propagated pressure waves traveled over significantly longer distances (4.5 to 6 cm) than those induced by acid infusion (3 cm). We conclude that the motor response to small amounts of intraduodenal nutrients and acid is dependent on the chemical composition of the stimulus. The findings suggest that chemoreceptors in the duodenal wall provide input to local or regional control mechanisms involved in the regulation of duodenal motility. PMID- 11478500 TI - Sulfated polysaccharides, but not cellulose, increase colonic mucus in rats with loperamide-induced constipation. AB - Colonic mucus is decreased in a rat model of spastic constipation, and some types of water-insoluble dietary fiber increase colonic mucus when consumed by rats for several weeks. However, little is known about the effect of water-soluble dietary fiber on the colonic mucus. The aim of the present study was to investigate the effect of various types of water-soluble dietary fiber on colonic mucus in a rat model of spastic constipation. Oral administration of 1.5 mg/day of carrageenan and chondroitin sulfate increased the fecal excretion, epithelial mucin production, thickness of the mucous layer, and amount of luminal mucus in loperamide-administered rats. Sodium alginate, 5 mg/day, thickened the mucus layer at the fecal surface. Cellulose, 5 mg/day, increased the fecal excretion but not the colonic mucus. Carrageenan, chondroitin sulfate, and sodium alginate, but not cellulose, increased colonic mucus in the rat model of spastic constipation. PMID- 11478501 TI - Phorbol ester treatment increases paracellular permeability across IEC-18 gastrointestinal epithelium in vitro. AB - The phorbol ester, TPA, transiently increases the transepithelial permeability across the gastrointestinal epithelium formed by IEC-18. There was a significant decrease in transepithelial resistance (R(T)) between 0 and 1.5 hr, accompanied by increased flux of polyethylene glycol (4000 MW), suggesting that the increase was across the tight junction. By 2 hr, the decrease in R(T) reversed and maintained control level. The transepithelial permeability increase was prevented by coincubation with the protein kinase C (PKC) inhibitor bisindolylmaleimide. There was a rapid (within 15 min) translocation of PKC-alpha from the cytosolic to the "membrane-associated" compartment, followed by a down-regulation that was detectable within 60 min of TPA treatment. The down-regulation of PKC-alpha from the membrane was prevented by either calpain inhibitor I or MG-132 and resulted in a sustained permeability increase. The permeability changes were not accompanied by significant effects on the amount or localization of the tight junctional proteins, occludin and ZO-1. However, occludin did show a reversible increase in phosphorylation with TPA treatment. Together these data support a role for PKC-alpha-mediated regulation of barrier permeability in an in vitro model of small intestinal epithelium, perhaps through modulation of the phosphorylation state of the tight junctional protein, occludin. PMID- 11478502 TI - High prevalence of celiac disease in Italian general population. AB - The worldwide increase of celiac disease prompted us to assess its prevalence in the Italian general population. The 3483 inhabitants of Campogalliano were tested for immunoglobulin A anti-endomysial antibodies. Twenty subjects showed antibody positivity and duodenal biopsy detected typical mucosal lesions of celiac disease in 17 of them; the remaining three cases had a normal villous architecture, but the finding of increased gamma/delta intraepithelial lymphocytes in all and the heterodimer DQA1*0501, DQB1*0201 in two of them was consistent with potential celiac disease. Only one patient had an overt malabsorption syndrome, characterized by diarrhea, weight loss, and severe weakness. In eight subjects atypical symptoms of celiac disease, such as dyspepsia and depression, were present, whereas the remaining subjects were silent. Celiac disease was more frequent in younger age groups. Our cross-sectional design study demonstrates that celiac disease prevalence in the Italian general population is 4.9 per 1000 (95% CI 2.8-7.8), increasing up to 5.7 per 1000 (95% CI 3.5-8.8) with the inclusion of potential cases. PMID- 11478503 TI - Celiac disease in patients with variegate porphyria. PMID- 11478504 TI - Improved parameters of lactose maldigestion using lactulose. AB - Lactulose is a disaccharide derived from lactose. There has been recent rekindling of interest in the possible benefits of pro- and prebiotics: mainly, lactic acid-producing bacteria and lactulose for the lower intestine. Since lactose maldigestion is a common genetic trait, we undertook this study to delineate similar effects between these two disaccharides. Nine healthy lactose maldigesting subjects underwent two separate periods of three weeks adaptation, first with 10 g twice daily lactulose and then 1.5 g twice daily lactose (in milk). Adaptation was defined by reduced breath Hydrogen (BH2) and symptoms after 50 g lactose challenges. In six subjects fecal beta-galactosidase was measured. All subjects consumed some lactose daily. In the first period, eight subjects improved symptoms and reduced BH2 significantly, while in the second period they did not. Fecal beta-galactosidase significantly increased after lactulose. This study supports the notion that lactulose and lactose may have similar clinical effects. PMID- 11478505 TI - Role of polymorphisms in the interleukin-10 gene in determining disease susceptibility and phenotype in inflamatory bowel disease. AB - Interleukin-10 (IL-10) has a key role in regulating mucosal inflammation. The role of functional polymorphisms at positions -627 and -1117 in the IL-10 gene as candidate susceptibility loci in inflammatory bowel disease and their importance in determining disease extent were evaluated in 159 patients with ulcerative colitis (83 left-sided; 76 extensive), 90 patients with Crohn's disease (22 small bowel; 29 large bowel; 39 both), and 227 controls. Genotyping was performed either by PCR-RFLP assays (-627 site) or SSCP analysis (-1117 site). An excess of -627A allele was observed in patients with left-sided colitis (52%) compared with controls (33%; P = 0.004) suggesting that IL-10 may influence the extent of the disease. These results were not replicated in a newly recruited group (N = 100) of patients with UC. We conclude that polymorphisms at -627 and -1117 sites in the IL-10 gene do not contribute to the susceptibility to IBD or determining the extent of the disease in our population. PMID- 11478506 TI - Lipopolysaccharide-induced gastroprotection is independent of the vagus nerve. AB - This study was done to examine the role of the vagus nerve in a model of gastric injury during endotoxemia. In conscious rats, lipopolysaccharide (LPS; 20 mg/kg i.p.) treatment for 5 hr prevented macroscopic gastric injury caused by acidified ethanol (150 mM HCl/50% ethanol). In addition, LPS enhanced gastric luminal fluid accumulation, decreased gastric mucosal blood flow (laser Doppler), and increased plasma gastrin levels (radioimmunoassay). Subdiaphragmatic truncal vagotomy, performed 7 days prior to LPS inhibited LPS-induced fluid accumulation, further reduced gastric mucosal blood flow following LPS, and augmented LPS-induced gastrin release compared to those in pyloroplasty controls. Atropine (1 mg/kg i.p.) prevented LPS-induced fluid accumulation but did not influence the effects of LPS on blood flow or gastrin release. Neither vagotomy nor atropine negated LPS-induced gastroprotection. This is the first report to examine the role of cholinergic nerves in the stomach during endotoxemia. The data indicate that LPS causes accumulation of gastric luminal fluid in part through its effects on cholinergic nerves. In contrast, the effects of vagotomy on blood flow and gastrin release following LPS involve a noncholinergic pathway. However, LPS induced gastroprotection is independent of the vagus nerve. PMID- 11478507 TI - Impact of a formulary change in proton pump inhibitors on health care costs and patients' symptoms. AB - Patients may fail to successfully undergo a switch in therapy associated with a formulary change. The aim of this study was to measure health care costs and outcomes among patients who failed a formulary change in proton pump inhibitors in a VA medical center. Patients who failed a switch from omeprazole to lansoprazole (N = 51) were matched with patients who were successfully switched (N = 51). Health care utilization data was gathered from VA electronic databases and medical records for six months before and after the switch and, for failure patients, during the lansoprazole trial period. Statistical comparisons between failure and success patients were performed on changes in health care costs between these time periods. Health outcome data for the lansoprazole trial period and subsequent omeprazole reinstatement period were obtained through a telephone questionnaire of failure patients. Changes in total health care utilization costs did not differ significantly between failure and success groups for any of the time periods. Failure patients had significantly poorer health outcomes during their lansoprazole trial periods with significantly greater severity of heartburn and severity and frequency of acid regurgitation (P < 0.001). In conclusion, the formulary change had a negative impact upon health outcomes among failure patients but did not significantly affect their health care utilization costs. Identification of failure patients early in their lansoprazole trial periods could improved their health outcomes and satisfaction with medical care. PMID- 11478508 TI - Symptoms associated with gastroesophageal reflux disease: development of a questionnaire for use in clinical trials. AB - Many persons who suffer from GERD report additional symptoms, e.g., chest pain, dyspepsia, dysphagia, that are often not measured in clinical trials even though they may be distressing to the GERD sufferer. The primary goal of this study was to develop and assess the psychometric characteristics of a new GERD symptom scale measuring frequency, severity, and distress. The GERD Symptom Assessment Scale (GSAS) was administered to a sample of 169 GERD sufferers at baseline and two weeks. Internal consistency, construct validity, and test-retest reliability were assessed. Responsiveness was evaluated using clinical trial data assessing drug efficacy. RESULTS: Internal consistency was >0.80 for the symptom severity and distress scales. All three scales showed stability over two weeks (ICC >0.70). Both validity hypotheses were supported. Comparison of effect sizes showed the GSAS is sensitive to changes in severity of symptoms. In conclusion, the GSAS is a reliable, valid, and responsive measure of GERD symptoms. PMID- 11478509 TI - H. pylori and functional dyspepsia: increased serum antibodies as an independent risk factor? AB - The relationship between H. pylori and functional dyspepsia is controversial. Hypothesizing that subjects with a more intense immune response to H. pylori (and hence higher antibody titers) would be at greater risk of dyspepsia, we aimed to identify risk factors for the development of dyspeptic symptoms. In all, 491 healthy blood donors with no history of peptic ulceration and 74 consecutive patients with a confirmed diagnosis of functional dyspepsia were studied. Symptoms and potential risk factors [nonsteroidal antiinflammatory drugs (NSAIDs), alcohol, and smoking] were measured by a validated questionnaire. H. pylori status was determined by IgG antibodies using a validated ELISA test with a cutoff titer for a positive serology of 10 units/ml. Logistic regression analysis assessed the association between risk factors and dyspepsia. Among blood donors, 21% (95% CI 17.6-24.8) reported dyspepsia (pain localized to the upper abdomen); 7.7% (95% CI 5.5-10.4) had frequent dyspepsia (>6 times in the prior year). The age-adjusted prevalence of H. pylori was not significantly different in blood donors with (39.5%, 95% CI 24.0-56.6) and without frequent dyspepsia (34.2%, 95% CI 29.8-38.36), but was significantly greater in patients with functional dyspepsia (68.8%, 95% CI 57.3-77.9). In the combined study population of blood donors and patients with functional dyspepsia, logistic regression adjusting for age identified the following independent risk factors for frequent dyspepsia: high serum antibody levels against H. pylori (OR for IgG titer >50 units/ml vs H. pylori titers 11-50 units/ml 4.6, 95% CI 2.7-7.8) and consumption of standard NSAIDs (OR 2.4,95% CI 1.3-4.5). In contrast, alcohol (OR 0.6, 95% CI 0.3-1.0), smoking (OR 1.5, 95% CI 1.0-2.3) or positive H. pylori serology with titers < or = 50 units/ml (OR 1.6, 95% CI 0.8-2.9) were not associated with frequent dyspepsia. In conclusion, in a subgroup of H. pylori-infected subjects who have high antibody titers, H. pylori appears to be associated with functional dyspepsia. PMID- 11478510 TI - Alterations of epithelial permeability by Helicobacter and IL-1beta in vitro: protective effect of rebamipide. AB - Infection with Helicobacter increases the transcellular passage of macromolecules across the epithelium, and this effect can be prevented by a gastroprotective agent rebamipide. The aim was to gain insight into the mechanisms involved. The HT29-19A intestinal epithelial cells grown on microporous filters as monolayers were incubated in the presence or absence of rebamipide (1 or 2 mM) with: (1) suspension of a wild H. pylori strain, (2) IL-1beta (0.5 ng/ml) + IFN-gamma (2 units/ml). After incubation, the monolayers were submitted to evaluation of apoptosis by using the apoptotic cell death detection ELISA kit and to assessment of epithelial permeability in Ussing chamber where the ionic conductance (G), fluxes of mannitol (J(Man)) and of horseradish peroxidase in both intact (J(HRPi))- and degraded (J(D)) form, were measured. H. pylori increased the intact HRP fluxes across the barrier (J(HRPi) = 17 +/- 20 vs 97 +/- 70 ng/hr/cm2, P < 0.007), an effect prevented by rebamipide (J(HRPi) = 33 +/- 34 ng/hr/cm2, P < 0.006). IL-1beta increased the ionic conductance (G = 5.5 +/- 1.0 and 21.0 +/- 7.0 mS/cm2, P < 0.006), the intact HRP fluxes (J(HRPi) = 18 +/- 15 and 476 +/- 344 ng/hr/cm2, P < 0.006), and the apoptotic index of the cells (AI = 1 +/- 0 vs 3.7 +/- 0.8), all effects prevented by rebamipide (G = 12 +/- 4.9 mS/cm2, J(HRPi) = 79 +/- 38, AI = 1.6 +/- 0.6, P < 0.03 as compared to IL-beta-treated cells). In basal conditions, rebamipide increased the integrity of the barrier (G = 7.5 +/- 2.3 vs 6.0 +/- 1.8 mS/cm2 for controls, P < 0.007). In conclusion, H. pylori as well as IL-1beta, may alter epithelial permeability and rebamipide may exert its protective effect on gastric mucosa by reinforcing the epithelial barrier in normal conditions and by counteracting the deleterious effect of Helicobacter pylori and IL-1beta on macromolecular permeability. PMID- 11478511 TI - Effectiveness of acid suppression in preventing gastroesophageal reflux disease (GERD) after successful treatment of Helicobacter pylori infection. AB - There is evidence that Helicobacter pylori eradication might predispose to gastroesophageal reflux disease (GERD). The aim of this prospective study was to examine the effectiveness of antisecretory treatment, after successful H. pylori eradication, in preventing GERD, since no data exist so far. Eighty initially H. pylori(+) patients, without GERD at the time of H. pylori eradication [50 peptic ulcer (PU) and 30 nonulcer (NU), 55 men, 25 women, median age 38 years, range 19 57], after successful H. pylori eradication were randomized to recieve either omeprazole 20 mg daily (group A) or no treatment (group B) for one year. All patients underwent upper gastrointestinal endoscopy at 0, 6, and 12 months or when GERD symptoms occurred. There were 40 patients in each group, and there were no statistically significant differences between the two groups in terms of sex, age, body weight, ulcer/no ulcer ratio, and other demographic data. Seven patients from group A and five patients from group B were lost to follow-up, and therefore there were 33 and 35 patients in groups A and B, respectively, who completed the study. One of 33 patients in group A (3%) and 10/35 (28.5%) in group B developed GERD symptoms during follow-up (P = 0.0022). The respective values for esophagitis were 0/33 (0%) and 6/35 (17.1%) (P = 0.0083). In conclusion, antisecretory treatment in H. pylori(+) patients, after successful eradication, is effective in preventing GERD. PMID- 11478512 TI - Helicobacter pylori infection and gastric function in patients with fundic atrophic gastritis. AB - In the present study we evaluated the relation among histology, H. pylori, IgG to H. pylori, gastric emptying, and acid secretion in 43 patients with fundic atrophic gastritis. On the basis of gastric acid secretion, patients were divided into three subgroups: patients with preserved acid secretion (Group 1), patients with hypochlorhydria (Group 2), and patients with achlorhydria (Group 3). Fundic glandular atrophy was more severe in hypoachlorhydric patients than in those with preserved acid secretion (P < 0.05 vs Group 2, P < 0.005 vs Group 3). H. pylori colonization was found in 94% of patients in Group 1, in 61% of patients in Group 2, and in only 8% of patients in Group 3 (P < 0.001 vs Group 1, P < 0.05 vs Group 2). Conversely, serological positivity to H. pylori was high in all three subgroups of patients (100% in Group 1, 77% in Group 2, 92% in Group 3). Gastric emptying was delayed in atrophic patients, particularly in those with hypoachlorhydria. Our data suggest that fundic atrophic gastritis represents a possible end stage of H. pylori infection, characterized by a progressive disappearance of the bacterium and a progressive deterioration of gastric functions. PMID- 11478513 TI - Parathyroid hormone-related protein production by adenocarcinoma in Barrett's esophagus patient with dermatomyositis. PMID- 11478514 TI - Dietary organic chromium supplementation and its effect on the immune response of rainbow trout (Oncorhynchus mykiss). AB - Recently, there has been an increasing interest in the potential effect of dietary chromium on the health of fish, particularly with respect to their metabolism and growth. Information as to the role of this mineral on their immune response, is limited however, so the aim of this study was to assess the effects of dietary chromium yeast supplementation on the immune response of rainbow trout (Oncorhynchus mykiss). Juvenile rainbow trout (56 g average weight) were fed three semipurified diets containing different levels of chromium (1540, 2340 and 4110 ppb), obtained by supplementing a basal diet with 800 or 2570 ppb chromium yeast, for 6 weeks. After this, time differences in their immune response were examined. A positive influence was observed on serum lysozyme activity at this time in fish maintained on the high chromium diet. The respiratory burst of head kidney macrophages was also examined, and statistical differences were found in the level of respiratory burst elicited by macrophages from both groups of fish fed supplemented chromium after 3 and 6 weeks of feeding (absorbance at 3 weeks: 0.118, 0.166. 0.151 and 6 weeks 0.114, 0.168, 0.151 for the 1540, 2340 and 4110 ppb groups). Macrophages of fish receiving diets supplemented with chromium also had a greater ability to phagocytose yeast after 6 weeks than the control fish (40.5, 48 and 48.5% macrophages phagocytic in the 1540, 2340 and 4110 ppb groups, respectively). The results of the study show that chromium yeast is able to modulate the immune response of rainbow trout, and this effect appears to be both dose- and time-dependent. PMID- 11478515 TI - Viral haemorrhagic septicaemia virus induces vig-2, a new interferon-responsive gene in rainbow trout. AB - An mRNA differential display methodology was used to study the rainbow trout response to viral infection. A new transcript (vig-2) induced by viral haemorrhagic septicaemia virus (VHSV) in rainbow trout leucocytes was identified from the head-kidney. vig-2 was also induced in vivo during experimental infection and following DNA immunisation with a plasmid containing a gene encoding the viral glycoprotein. Viral induction of vig-2 was blocked by cycloheximide (CHX), indicating its dependency on a newly synthesised intermediate protein. This intermediate protein is most probably related to interferon because treatment of cells with a conditioned medium displaying an interferon-like activity resulted in a strong vig-2 expression, which was not blocked by CHX treatment. The cDNA sequence of the vig-2 transcript displays several mRNA destabilisation motifs and two signals characteristic of immediate early gene expression. Curiously, vig-2 has no evident encoding potential except for a small 51 amino acid putative polypeptide with no clear similarity to any sequence available in the databanks. Therefore, the complete vig-2 genomic sequence was determined from a lambda phage clone retrieved from a genomic DNA library of rainbow trout. The genomic organisation of vig-2 shows five exons delimited with typical splice acceptor and donor sites. A promoter with a canonical ISRE, confirming that vig-2 is an interferon-responsive gene, is also present 115 nt upstream of the first exon. PMID- 11478516 TI - Immunoglobulin genes and antibody responses in the spotted wolffish (Anarhichas minor Olafsen). AB - The spotted wolffish Anarhichas minor Olafsen is a promising new species in aquaculture in the cold waters of northern Norway. In this paper, some basic immunological studies of this marine species are reported. Of comparative interest are the cDNA sequences of the immunoglobulin transcript and the antibody responses to model antigens. Of more practical importance are the humoral immune responses and antibody specificities to potentially pathogenic bacteria. Full length cDNA clones encoding the immunoglobulin heavy and light chains in the spotted wolffish were sequenced demonstrating variable degrees of similarity to other teleost fish species. Also in the spotted wolffish the CH4 domain was deleted in the transmembrane form of the immunoglobulin heavy chain (IgH) as a receptor on B cells, with the transmembrane exon spliced directly to the CH3 domain. The antibody responses to various antigens like hapten-carrier molecules, protein antigens and bacterial pathogens were relatively high, but with some interesting exceptions. Anti-hapten responses to NIP and FITC were high while anti-DNS responses were low, but more surprisingly, there was hardly any B-cell response to the carrier molecule LPH. On the other hand, protein antigens like CGG and BSA were highly immunogenic in the spotted wolffish as were the bacterial antigens Vibrio anguillarum, V. salmonicida and Aeromonas salmonicida. PMID- 11478517 TI - In vitro interactions between rainbow trout (Oncorhynchus mykiss) macrophages and Vibrio anguillarum serogroup O2a. AB - The sensitivity of Vibrio anguillarum serogroup O2a to killing by rainbow trout macrophages in the presence or absence of specific antibodies and complement components was evaluated using an in vitro assay. Fluorescence microscopy revealed that V. anguillarum serogroup O2a was phagocytosed by rainbow trout macrophages. In the absence of specific antibodies and complement components the bacteria were killed to a limited extent by the macrophages and there was no increased killing if the bacteria were opsonised with either antibodies or antibodies and complement. Furthermore, activated macrophages did not show enhanced ability to kill the bacteria. Vibrio anguillarum serogroup O2a were susceptible to both cell-free superoxide anion (O2-) and hydrogen peroxide (H2O2), which might be generated during the macrophage respiratory burst and the bacteria did not quench cell-free O2-. However, the production of O2- by macrophages was undetectable during the first 30 min following infection and no respiratory burst was inducible by phorbol myristate acetate (PMA) 4 h after infection with V. anguillarum. This suggests that the bacteria were able to inhibit the production of O2- by the infected macrophages. Naive fish were protected when passively immunised with anti-V. anguillarum serogroup O2a antiserum. However, previous results suggest that antibodies are unlikely to provide the fish with protective immunity directly through activation of the complement system and lysis of the bacterial cells. The present in vitro findings suggest that the protective mechanisms of antibody against V. anguillarum serogroup O2a may not involve the opsonising effect of antibodies for enhanced killing by macrophages. However, the possibility exists that such antibodies may prevent the attachment of the pathogen to the host's tissues. PMID- 11478518 TI - Rodlet cells and granulated leucocytes in the bulbus arteriosus of swordtail, Xiphophorus helleri L. and platy, Xiphophorus maculatus L. (Poecilidae: Teleostei). PMID- 11478519 TI - The gene is not dead, merely orphaned and seeking a home. AB - Despite announcements and obituaries, news of the death of the gene has been greatly exaggerated, or so says the gene as it struggles to survive and find a safe haven from which to steer its course through development and evolution. In this short piece, I consider recent claims that the gene is dead. I conclude that the gene is alive and well, living and functioning in the cell, which is both its natural home and a fundamental unit of evo-devo. PMID- 11478520 TI - Microevolutionary analysis of the nematode genus Pristionchus suggests a recent evolution of redundant developmental mechanisms during vulva formation. AB - To identify the mechanisms by which molecular variation is introduced into developmental systems, microevolutionary approaches to evolutionary developmental biology have to be taken. Here, we describe the molecular and developmental characterization of laboratory strains of the nematode genus Pristionchus, which lays a foundation for a microevolutionary analysis of vulva development. We describe 13 laboratory strains of the Pristionchus genus that are derived from natural isolates from around the world. Mating experiments and ITS sequence analysis indicated that these 13 strains represent four different species: the gonochoristic species P. Iheritieriand three hermaphroditic species, P. pacificus, P. maupasi, and an as yet undescribed species Pristionchus sp., respectively. P. pacificus is represented by five different strains isolated from California, Washington, Hawaii, Ontario, and Poland. Developmental differences during vulva formation are observed between strains from different species but also between strains of P. pacificus, like the strains from California and Poland. In particular, redundant developmental mechanisms present during vulva formation in P. pacificus var. California are absent in other strains. Amplified restriction fragment length polymorphism (AFLP) analyses of the P. pacificus strains revealed that the American strains are highly polymorphic. In contrast, the developmentally distinct strain from Poland is identical to the Californian strain, suggesting that the developmental differences rely on a small number of changes in developmental control genes rather than the accumulation of changes at multiple loci. PMID- 11478521 TI - Conservation of a DPP/BMP signaling pathway in the nonbilateral cnidarian Acropora millepora. AB - Members of the TGF-beta superfamily of signaling molecules are widespread in metazoans, but the evolutionary origin of particular subclasses of signaling mechanisms is poorly defined. The DPP/BMP class, for example, is implicated in dorsal-ventral patterning, neural patterning, and limb development. Here we report the presence of several components of a DPP/BMP-specific signal transduction cascade in a nonbilateral animal, the coral Acropora millepora. The discovery of these components, a putative type I receptor and two putative receptor-activated Smads, suggests that DPP/BMP signaling predates both dorsal ventral pattern formation and limb development. We postulate that an ancestral role in neuroepithelial patterning may account for the high level of conservation between DPP/BMP signaling components found in this nonbilateral animal and the more complex triploblastic organisms of the arthropod and chordate phyla. PMID- 11478522 TI - Divergent structure and function of the bicoid gene in Muscoidea fly species. AB - We have investigated the evolution of the bicoid (bcd) gene in fly species of the Muscoidea Superfamily. We obtained the complete bcd sequence from the housefly Musca domestica and found polymorphism in the coding region among Musca strains. In addition to Musca, we cloned most of the bcd coding sequences from two blowfly species Calliphora vicina and Lucilia sericata. The 5' and 3' regulatory regions flanking the Musca bcd gene are widely diverged in sequence from Drosophila; however, some important sequence motifs identified in Drosophila bcd are present. The predicted RNA secondary structures of the 3' UTRs are similar, despite sequence divergence. Comparison of Bicoid (Bcd) proteins shows a serine-rich domain of unknown function is present in the Muscoidea species, but is absent in other species. The in vivo function of bcd in Musca was tested by RNAi to mimic loss of function phenotype. We obtained a head defect phenotype similar to weak bcd alleles of Drosophila. Although our comparisons initially suggest functional conservation between species, closer inspection reveals significant differences. Divergence of structural motifs, such as regulatory elements in flanking regions and conservation of protein domains in some species but not in others, points to functional divergence between species. We suggest that the larger embryonic size in Muscoidea species restricts the morphogenetic activity of a weak Bcd activator, which has evolved a more specialized role in head determination and lost some functions in thoracic development. PMID- 11478523 TI - Sipunculan ParaHox genes. AB - Our perspective on the origin and evolution of the Hox gene cluster changed with the discovery of the ParaHox gene cluster in amphioxus (Cephalochordata; Branchiostoma floridae) (Brooke et al. 1998). The ParaHox gene cluster contains three homeobox genes (Gsx, Xlox, Cdx) and is deduced to be a paralogue (evolutionary sister) of the Hox gene cluster. If this deduction is correct, animals with Hox genes should also possess ParaHox genes. Paradoxically, however, only deuterostome animals have thus far been shown to contain all three ParaHox genes. Here we report the cloning of all three ParaHox genes from each of two species within the phylum Sipuncula. This is the first demonstration of all three ParaHox genes in the genome of a protostome animal and confirms that the common ancestor of protostomes and deuterostomes possessed all three ParaHox genes. Furthermore, it implies that the ParaHox genes are of sufficient functional importance in both protostomes and deuterostomes that they have all been conserved in both of these bilaterian clades. PMID- 11478524 TI - Developmental drive: an important determinant of the direction of phenotypic evolution. AB - Over any period of evolutionary time, the prevailing ontogenetic trajectory within a lineage may either recur unchanged from generation to generation (stasis) or alter (developmental reprogramming). A key question about reprogramming is whether it exhibits intrinsic biases in favor of some sorts of change and against others, which may be referred to respectively as "drive" and "constraint." A simple logical argument suggests that both drive and constraint should be common, and conversely that cases of equiprobable modification in various phenotypic directions should be relatively rare. These proposals, that drive and constraint exist and that they are common, appear to be widely accepted, even among neo-Darwinians, who are sometimes portrayed as rejecting them. What is more controversial is that developmental drive (and constraint) can have a powerful influence on the direction of evolutionary change. It is argued that such an influence will occur, and indeed may be pervasive. PMID- 11478525 TI - How many processes are responsible for phenotypic evolution? AB - In addressing phenotypic evolution, this article reconsiders natural selection, random drift, developmental constraints, and internal selection in the new extended context of evolutionary developmental biology. The change of perspective from the "evolution of phenotypes" toward an "evolution of ontogenies" (evo-devo perspective) affects the reciprocal relationships among these different processes. Random drift and natural selection are sibling processes: two forms of post-productional sorting among alternative developmental trajectories, the former random, the latter nonrandom. Developmental constraint is a compound concept; it contains even some forms of natural ("external" and "internal") selection. A narrower definition ("reproductive constraints") is proposed. Internal selection is not a selection caused by an internal agent. It is a form of environment-independent selection depending on the level of the organism's internal developmental or functional coordination. Selection and constraints are the main deterministic processes in phenotypic evolution but they are not opposing forces. Indeed, they are continuously interacting processes of evolutionary change, but with different roles that should not be confused. PMID- 11478526 TI - Interpreting the homeobox: metaphors of gene action and activation in development and evolution. AB - Despite countless research efforts to demonstrate the precise developmental and evolutionary nature of homeobox genes, we are far from consensus on the role of this class of genes in development and evolution. This essay attempts to clarify the debate and to nip some problematic interpretations in the bud, by exploring metaphors of homeobox gene function in development and evolution. PMID- 11478527 TI - Do geneticists need Babel fish? PMID- 11478528 TI - Health Insurance Portability and Accountability Act is here: what price privacy? PMID- 11478529 TI - Considerations for a multiaxis nomenclature system for medical genetics. PMID- 11478530 TI - Inheritance of two HFE mutations in African Americans: cases with hemochromatosis phenotypes and estimates of hemochromatosis phenotype frequency. AB - PURPOSE: Two unrelated African Americans had hemochromatosis phenotypes and genotypes. We sought to identify origins of their HFE mutations and estimate frequencies of similar cases. METHODS: HFE and HLA genotyping were performed in index cases and family members. HFE genotypes of 1,373 African American controls in five regions were tabulated. RESULTS: Index cases had C282Y/C282Y and C282Y/H63D, respectively; each corresponding Ch6p was likely of Caucasian origin. In controls, frequencies of hemochromatosis-associated genotypes were as follows: C282Y/C282Y, 0.00011; C282Y/H63D, 0.00067; and H63D/H63D, 0.00101. CONCLUSIONS: Penetrance-adjusted estimates indicate that approximately 9 African Americans per 100,000 have a hemochromatosis phenotype and two common HFE mutations. Hemochromatosis-associated genotype frequencies varied 11.7-fold across regions. PMID- 11478531 TI - Favorable attitudes toward testing for chromosomal abnormalities via analysis of fetal cells in maternal blood. AB - PURPOSE: The NICHD Fetal Cell Isolation Study (NIFTY) was a multicentered project to isolate fetal cells from maternal blood to detect fetal chromosomal abnormalities. The project included a psychosocial component, which is the basis of this article. We examined the attitudes of high-risk pregnant women toward the availability of a maternal blood test to identify fetal chromosomal abnormalities, how women would respond to hypothetical normal and abnormal maternal blood testing results, and the factors associated with a woman's preference to have an invasive procedure in response to a normal maternal blood test. METHODS: High-risk pregnant women (N = 854) planning to have prenatal diagnostic invasive testing (amniocentesis or chorionic villus sampling) completed a survey. RESULTS: The women highly favored maternal blood testing. Almost all women would seek invasive testing after an abnormal blood test. Only half of the women would seek invasive testing after a normal blood test; these women were older, more willing to terminate their pregnancy, and valued the increased accuracy of invasive testing more highly than women who would not have invasive testing after a normal maternal blood test. CONCLUSIONS: Women having invasive diagnostic testing welcome a noninvasive procedure that uses fetal cells in maternal blood, and its availability would decrease invasive testing by approximately 50%. Research needs to examine the attitudes and anticipated responses of other risk groups as well as the effects of information about maternal blood test sensitivity and specificity on attitudes and responses. PMID- 11478532 TI - Townes-Brocks syndrome versus expanded spectrum hemifacial microsomia: review of eight patients and further evidence of a "hot spot" for mutation in the SALL1 gene. AB - PURPOSE: It can be difficult to differentiate clinically between hemifacial microsomia (HFM) and Townes-Brocks syndrome (TBS). The distinction is important because TBS is inherited as an autosomal dominant trait, whereas HFM is sporadic. METHODS: We performed a retrospective analysis of eight patients with HFM expanded spectrum and anal anomalies to determine whether this subset has TBS. RESULTS: Two patients had major phenotypic findings of TBS. Sequencing of SALL1, the gene mutated in TBS, in four of the eight patients revealed one with a C --> T transition (resulting in a nonsense mutation R276X) at a previously identified mutational "hot spot." CONCLUSION: Patients with overlapping features of both syndromes should be screened for SALL1 mutations. PMID- 11478533 TI - Packed red cell transfusion does not compromise chromosome analysis in newborns. AB - PURPOSE: Critically ill neonates are frequently transfused with packed red cells. Some of these transfused neonates also need chromosome analysis. There is a long standing tradition in pediatrics of not performing chromosome analysis after transfusion. We wished to determine whether transfusion with packed red cells affect the cytogenetic results in neonates. METHOD: The medical records of all neonates at the Medical College of Georgia who had had chromosome analysis between June 1995 and June 1998 were reviewed. Ten neonates had received transfusion prior to cytogenetic testing. Of these 10 infants, two had been transfused two or more times. Routine cytogenetic analysis of 20 metaphases at 550-band level had been performed on all 10 patients. Heteromorphic markers were compared in 10 randomly selected metaphases for any discrepancy. To determine whether there were theoretical reasons to delay chromosome analysis in transfused neonates, samples of irradiated, and/or filtered, and nonfiltered blood were obtained from the blood bank and analyzed for the presence of lymphocytes. RESULTS: Prior transfusion did not affect karyotype results. A nonmosaic abnormal karyotype was found in 3 of the 10 patients. A fourth patient's karyotype was 45,X/47,XXX. This mosaicism was constitutive and consistent as demonstrated by a follow-up chromosome analysis. All other abnormal karyotypes were consistent with the dysmorphic phenotype. Randomly selected metaphases did not show any differences in the identifiable heteromorphic markers in all 10 patients. Although there was a 50% chance of patients receiving blood from a donor of opposite sex, there were no instances in which cells with a karyotype of the opposite sex were found in the patients' blood. The irradiated and filtered cultured donor blood samples did not show any metaphases. However, metaphases were seen in the cultures from nonfiltered and nonirradiated donor blood. CONCLUSIONS: Based on these results one does not need to delay karyotyping babies who have had blood transfusions. Packed red cell transfusion in newborns does not compromise the accuracy of chromosome analysis in our study even with multiple transfusions. PMID- 11478535 TI - Advocating for genetics at the local level: the Medicare Part B Carrier Advisory Committee. PMID- 11478534 TI - Suspected gonadal mosaicism for isochromosomes 18p and 18q unsubstantiated by fluorescence in situ hybridization analysis of sperm. AB - PURPOSE: A father had two children, one with isochromosome 18p, and another with isochromosome 18q. The father was counseled that he might have gonadal mosaicism for isochromosomes 18p and 18q, which could confer a high recurrence risk. METHODS: A sperm sample from the father was analyzed with fluorescence in situ hybridization probes for 18p and 18q. RESULTS: More than 1,000 sperm were scored and none were found with two 18p or 18q signals. There were no differences in the father's specimen compared to a control. CONCLUSIONS: There was no evidence for gonadal mosaicism. It is important to confirm clinical hypotheses whenever possible. PMID- 11478536 TI - Feminism meets the "new" epidemiologies: toward an appraisal of antifeminist biases in epidemiological research on women's health. AB - This essay explores an alternative paradigm for epidemiology, one which is explicitly informed by a feminist perspective. We intend to expand upon recent critiques and debates within the emergent fields of "critical", "popular", and "alternative" epidemiology to examine how epidemiology's conceptual models--which are meant to contribute to the prevention of social inequalities in health, but may instead reinforce social hierarchies based on gender, race, and class- constrain our understanding of health and disease. Specifically, we examine persistent antifeminist biases in contemporary epidemiological research on women's health. Issues highlighted include: problem definition and knowledge production in women's health: biological essentialization of women as reproducers; and decontextualization and depoliticization of women's health risks. As part of this critique, we include suggestions for an emancipatory epidemiology that incorporates an alternative feminist framework. PMID- 11478537 TI - Comorbid depressive symptomatology: isolating the effects of chronic medical conditions on self-reported depressive symptoms among community-dwelling older adults. AB - Advances in medical technology and surgical knowledge have greatly extended the life expectancy of older individuals with chronic disabilities. Among the older adult population the prevalence of comorbid chronic illness and depressive symptoms has often been investigated. Yet there continues to be a lack of understanding about the consequences of specific chronic illnesses on depressive symptoms. Using cross-sectional data while simultaneously controlling the effects of various socioenvironmental, demographic, and other factors, this study analyzed the prevalence of self-reported depressive symptoms in relation to chronic illness. The cumulative effect of medical comorbidity and the specific effect of individual chronic illnesses were examined. The sample consisted of 359 older white Americans aged 55-93. Overall, the findings show that self-reports of depressive symptoms were greater among those individuals who also reported digestive disorders, respiratory ailments, and heart problems. These results suggest that for older white Americans these three chronic conditions may present a greater mental health burden than other chronic illnesses. PMID- 11478538 TI - Neighbourhood and family influences on the cognitive ability of children in the British National Child Development Study. AB - This paper investigates the association between family poverty, the level of deprivation in electoral wards and children's cognitive test scores using data from the second generation in the 1991 sweep of the British National Child Development Study (1958 birth cohort). Family poverty has a significant association with lower test scores in children of all ages (4-18 years). Neighbourhood poverty has a significant association with lower test scores in children aged 4-5 years which, though somewhat attenuated, is independent of other socioeconomic indicators. Among children aged between 6 and 9 years, the association with neighbourhood deprivation is statistically accounted for by individual characteristics. Among children aged between 10 and 18 years, levels of neighbourhood deprivation were for the most part statistically insignificant. The family poverty--test score association among children aged between 10 and 18 years was mediated by the home environment. Mediated effects were stronger for family poverty--test score associations than for neighbourhood poverty. The use of a neighbourhood-level exposure related to the social environment leads to an understanding of the social determinants of children's outcomes that is more than the sum of individual and family-level measures. However, the size of the estimated effects of neighbourhood conditions is much smaller than the estimated effects of family-level conditions. Thus, it appears that families still should be viewed as the key agents in promoting positive development in children. PMID- 11478539 TI - Anxious adults vs. cool children: children's views on smoking and addiction. AB - Tobacco addiction represents a major public health problem, and most addicted smokers take up the habit during adolescence. We need to know why. With the aim of gaining a better understanding of the meanings smoking and tobacco addiction hold for young people, 85 focused interviews were conducted with adolescent children from economically deprived areas of Northern Ireland. Through adopting a qualitative approach within the community rather than the school context, the adolescent children were given the opportunity to freely express their views in confidence. Children seem to differentiate conceptually between child smoking and adult smoking. Whereas adults smoke to cope with life and are thus perceived by children as lacking control over their consumption, child smoking is motivated by attempts to achieve the status of cool and hard, and to gain group membership. Adults have personal reasons for smoking, while child smoking is profoundly social. Adults are perceived as dependent on nicotine, and addiction is at the core of the children's understanding of adult smoking. Child smoking, on the other hand, is seen as oriented around social relations so that addiction is less relevant. These ideas leave young people vulnerable to nicotine addiction. It is clearly important that health promotion efforts seek to understand and take into account the actions of children within the context of their own world-view to secure their health. PMID- 11478540 TI - Sense of coherence and school-related stress as predictors of subjective health complaints in early adolescence: interactive, indirect or direct relationships? AB - The role of sense of coherence (SOC) on the relationship between adolescent school-related stress and subjective health complaints was tested with structural equation modelling. As part of the crossnational WHO-survey 'Health behaviour in school-aged children 1997/98' Norwegian representative samples of 1592 grade 6, 1534 grade 8, and 1605 grade 10 students completed measures on SOC, school related stress and subjective health complaints. A test of nested structural models revealed that both stress-preventive (delta chi2 814. 86, p<0.001), stress moderating (delta chi2 11.74, p<0.02) and main health-enhancing (delta chi2 1289.1, p < 0.001) effects of SOC were consistent with the data. A model including all these relationships fitted the data well (CFI = 0.91, RMSEA = 0.04). Age-group comparisons revealed that the association between SOC and stress grew weaker with age (p<0.05), whereas the direct association between SOC and health complaints grew stronger (p<0.001). The main effect of SOC accounted for between 39% (11 year olds) and 54% (15 year olds) of the variance in subjective health complaints. Findings indicate that SOC may potentially be a salutogenic factor in adolescents' adaptation to school-related stress, and that relationships between SOC and healthy adaptation, may be evident in younger age groups than previously anticipated. PMID- 11478541 TI - Living a 'normal' life: young people coping with thalassaemia major or sickle cell disorder. AB - This qualitative paper explores the strategies and resources young people use to cope with sickle cell disorder or thalassaemia major, two haemoglobin disorders with serious implications for health and survival. By focusing on coping strategies, we explore how young people attempt to take control over their lives. The respondents, largely of South Asian and African Caribbean origin, aged between 10 and 19 years, valued maintaining a normal' life and struggled to achieve this normalcy. Strategies were employed to minimise difference from peers but these strategies remained vulnerable. Coping occurred in a dynamic space, involving negotiation and engagement with both personal and structural factors. Threats to normalcy did not always reside in the condition; life transitions, changes in social relationships and racist. disablist or sexist marginalisation also threatened coping strategies. PMID- 11478542 TI - Down's syndrome: cost, quality and value of life. AB - Routine prenatal screening is based on the assumption that it is reasonable for prospective parents to choose to prevent a life with Down's syndrome. This paper questions whether Down's syndrome necessarily involves the costs, limitations and suffering which are assumed in the prenatal literature, and examines the lack of evidence about the value and quality of life with Down's syndrome. Tensions between the aims of prenatal screening policies to support women's personal choices, prevent distress, and reduce the suffering and costs of disability, versus the inadvertent effects of screening which can undermine these aims, are considered. Strengths and weaknesses in medically and socially based models of research about disability, and their validity and reliability are reviewed. From exploratory qualitative research with 40 adults who have congenital conditions which are tested for prenatally, interviews with five adults with Down's syndrome are reported. Interviewees discuss their relationships, education and employment, leisure interests, hopes, aspects of themselves and of society they would like to change, and their views on prenatal screening. They show how some people with Down's syndrome live creative, rewarding and fairly independent lives, and are not inevitably non-contributing dependents. Like the other 35 interviewees, they illustrate the importance of social supports, and their problems with excluding attitudes and barriers. Much more social research with people who have congenital conditions is required, if prenatal screening policies and counselling are to be evidence based. PMID- 11478543 TI - Closing medical encounters: two physician practices and their implications for the expression of patients' unstated concerns. AB - When patients visit primary-care physicians, they frequently have more than one concern. Patients' first concerns are solicited by physicians at the beginnings of encounters. A challenge to health care is how to get patients' additional concerns raised as topics of discussion. If patients' additional concerns are addressed, it tends to occur at the end of encounters. Using the methodology of conversation analysis, this article identifies and describes the interactional organization of two physician-initiated communication practices that are used to negotiate the closure of the business of encounters and a transition into the activity of closing encounters themselves. These practices have different implications for the topicalization of patients' additional concerns. PMID- 11478544 TI - Male physicians' narratives about being in ethically difficult care situations in paediatrics. AB - This study is a part of a comprehensive investigation into the narratives of male and female physicians and nurses concerning their experience of being in ethically difficult care situations in paediatrics. Seventeen male physicians with a range of levels of expertise, working on various wards in paediatric clinics at two university hospitals in Norway, narrated 78 stories. The transcribed interview texts were subjected to hermeneutic analysis. All the interviewees related problems in both an action and a relation ethics perspective. The main focus was on ethical problems concerning life and death decisions. The central theme was overtreatment, which they felt they could easily slip into because of a lack of exact knowledge about the outcome of life-saving treatment. The less experienced physicians required criteria and ethical guidelines that could tell them when to stop treatment, and they expected the more experienced physicians to be able to teach them about such things. The more experienced physicians, however, told a different story about life-saving practices in paediatrics. They spoke of a very demanding life situation. In such a situation communication seems decisive, not only to clarify treatment questions, but primarily to cope with life. The physicians' main concern seemed to be the parents rather than the children. Both groups underlined the importance of professional distance to patients and to ethical problems. PMID- 11478545 TI - Taking responsibility for cancer treatment. AB - One hundred and fourteen consecutive patients with early breast cancer were entered into a study on the psychological effects of involvement in treatment choice. All women were offered counselling throughout. One group of women (n = 34), were advised to undergo mastectomy, due to the nature or position of the tumour. These women fared less well psychologically when compared on a battery of measures, before and after surgery, with women who were involved in choosing their own treatment (n = 80). The latter group itself was randomly allocated into two groups for taking explicit responsibility for treatment choice, using a double-blind procedure. These were a Patient Decision Group (n = 41) and a Surgeon Decision Group (n = 39). Results support the hypothesis that over and above the benefits of receiving their preferred treatment, women can further benefit from taking explicit responsibility for their treatment choice. PMID- 11478546 TI - Declining the offer of flexible sigmoidoscopy screening for bowel cancer: a qualitative investigation of the decision-making process. AB - Qualitative methods were used to investigate decision-making among a group of older adults who declined the offer of flexible sigmoidoscopy screening for bowel cancer. Interviews were conducted with 60 people (30 men and 30 women) who either had not responded to the screening letter or who responded saying that they were not interested in participating. The findings suggest that low perceived susceptibility to bowel cancer, in terms of current health status, family history or absence of bowel symptoms. was an important factor in the decision to decline screening. Procedural barriers such as embarrassment, pain/discomfort and perceived unpleasantness of the test were reported as relatively minor, although the test was considered more physically intrusive than other screening tests. Avoidant attitudes emerged as an important theme and were reported by a third of respondents. Distinct patterns of decision-making were also observed and three groups emerged from accounts: (i) forgetting or avoiding making a decision about the test (ii) a confident rejection of the test based on a few salient factors, and (iii) a more careful consideration of the test focusing on issues of susceptibility. The findings are discussed in the context of models of health behaviour and bowel cancer screening participation research. PMID- 11478547 TI - Spine fusion and bone grafting. PMID- 11478548 TI - Bone transplantation: where do we go from here? PMID- 11478549 TI - Radiologic case study. Pigmented villonodular synovitis. AB - Pigmented villonodular synovitis is a benign, locally aggressive disorder characterized by a unique clinical radiographic, and histopathologic presentation. While it is considered to be a rare disorder, a delay in diagnosis and treatment can potentially result in severe disruption of joint function due to subchondral invasion. Once the diagnosis is confirmed, treatment should consist of complete synovectomy. Recurrence is common, but malignant transformation is rare. The etiology of PVNS is still unknown, and perhaps its future discovery will assist in the definitive treatment of this disorder. PMID- 11478550 TI - Proximal rectus femoris lengthening. AB - There are two main problems with muscle tendon lengthening: thinness created by the technical procedure weakens the tendon, and in short tendons, lengthening may not provide enough length for the amount of joint motion required. In proximal rectus femoris lengthening, the distal ends of the reflected and straight heads are sutured together. Using this technique, the tendon will not be thinned, and a later second lengthening is possible. PMID- 11478551 TI - Safety of a clinical surveillance protocol with 3- and 6-week warfarin prophylaxis after total joint arthroplasty. AB - The charts of 1869 patients were reviewed for the occurrence of deep venous thrombosis (DVT) and pulmonary embolism after total hip or knee arthroplasty. Prophylaxis consisted of 3 (group 1; n=1235) or 6 (group 2; n=634) weeks low-dose warfarin, pneumatic compression boots worn by patients in the hospital, mobilization on the first postoperative day, and a clinical surveillance protocol. Venous ultrasound or ventilation/perfusion lung scintigraphy (V/Q) was performed only if patients became symptomatic. patients. Twenty-three (1.8%) patients were positive for DVT. Ventilation/perfusion lung scintigraphy was performed on 25 patients, and 5 (0.4%) patients were positive for pulmonary embolism. In group 2, 117 patients were evaluated for DVT, and 19 (3%) patients had positive results determined by ultrasound. Twenty-five patients were evaluated with V/Q and only 1 (0.16%) patient was positive for pulmonary embolism. No patient developed a fatal pulmonary embolism or postphlebitic syndrome. This prophylaxis protocol is an efficient and cost-effective method for the prevention of significant events after surgery. PMID- 11478552 TI - Arthroscopic rotator cuff interval capsular closure. AB - Closure of the rotator cuff interval is an important component of open stabilization techniques in shoulder surgery. This study describes a technique in which the deep layer of the capsule within the rotator cuff interval is closed arthroscopically. The effect of closure of this capsule within the rotator cuff interval on glenohumeral motion also is quantified. Sutures were placed from the superior glenohumeral ligament to the middle glenohumeral ligament and tied intra articularly in fresh-frozen cadaveric shoulders. Closure of the interval capsule resulted in statistically significant limitation of humeral elevation, external rotation, and extension. Arthroscopic closure of the deep layer of the rotator cuff interval capsule produced a visible superior shifting of the middle and inferior glenohumeral ligaments and imbricated the anterosuperior capsule of the shoulder. In addition, this procedure can be performed in conjunction with arthroscopic capsulolabral reconstruction. PMID- 11478553 TI - Pharmacokinetics of antibiotic prophylaxis in major orthopedic surgery and blood saving techniques. AB - The pharmacokinetics of cefuroxime, cefotiam, cefamandole, and ampicillin/sulbactam were randomly measured in 40 patients undergoing major orthopedic surgery associated with high blood and volume turnover and intraoperative blood salvage. Serum and bone concentrations and the pharmacokinetics occurring in the context of these procedures were measured. No changes in elimination half-life relative to a normal population occurred with cefuroxime, cefotiam, and ampicillin. Serum and tissue concentrations were slightly lower with cefamandole and sulbactam, but reapplication of the initial dose was required with all antibiotics 4 hours after the first application. PMID- 11478554 TI - Long-term local intensive preoperative chemotherapy and joint-preserving conservative surgery for osteosarcoma around the knee. AB - This study evaluated conservative joint-sparing surgery for patients with osteosarcoma around the knee. Of 23 patients with stage IIB osteosarcoma around the knee, 5 were treated with long-term (30-56 weeks) local intensive preoperative chemotherapy consisting of high-dose methotrexate, intra-arterial and intravenous cisplatinum, doxorubicin, and hyperthermic isolated regional perfusion. More conservative resection, sparing the knee joint, was performed with smaller sufficient surgical margin in these 5 patients, preserving good limb function. Excellent local effects were achieved in the resected specimens. These results suggest long-term local intensive preoperative chemotherapy, including intra-arterial cisplatin and hyperthermic isolated regional perfusion, help control local tumor and allow for more conservative surgery. PMID- 11478555 TI - Use of audiotapes for patient education, medical record documentation, and informed consent in lower extremity reconstruction. AB - Since 1992, the authors have audiotaped each new patient visit and provided a copy of that tape to the patient. In addition, an office copy of the entire patient interaction is maintained in an office file. One hundred twelve patients seen over a 6-month period participated in a survey measuring their satisfaction with this procedure, and six attorneys were surveyed after listening to a representative tape. Ninety percent of patients had positive comments about receiving the tape. Seventy percent played the tape for others, most commonly their spouses. The attorneys concluded the patient was well served by this process, and there were no increased malpractice issues or exposure. The cost of providing patients with the tapes is minimal, and the high level of patient satisfaction and increased understanding offer a considerable benefit to patients and office staff. PMID- 11478556 TI - A comparison of surgical and conservative treatment in ankle ligament tears. AB - Good results have been reported after both primary repair and conservative management of grade III ankle sprains. This prospective study found no advantage of operative treatment compared to taping; no significant differences were found between the groups with regard to objective or subjective stability, functional scores, or the overall result. PMID- 11478557 TI - Psoas hematoma presenting as hip pain. PMID- 11478558 TI - Genu varum affecting one identical twin. PMID- 11478559 TI - Iguana-transmitted Salmonella osteomyelitis. PMID- 11478560 TI - More on tension-band technique for ankle fusion. PMID- 11478561 TI - Subperiosteal osteoid osteoma of the distal phalanx of the fourth toe. PMID- 11478562 TI - Cervical spine injuries in athletes: current return-to-play criteria. PMID- 11478563 TI - Are physicians aware of the risks of alternative medicine? AB - Evidence supports the fact that alternative medical therapies play an increasingly prominent role in healthcare. Relevantly, this study posed three questions: (1) Do physicians ask their patients about their use of herbs/dietary supplements? (2) Do physicians use the available resources to evaluate the possible drug interactions and/or side effects of the dietary supplements? and (3) Are physicians aware of the side effects, drug interactions and contraindications of ten commonly used herbs? A questionnaire was randomly distributed to medical students and faculty of the State University of New York, Health Science Center at Brooklyn. One hundred sixty five surveys were returned out of 193 handed out (85%). Analysis revealed that although many physicians asked their patients about their use of alternative remedies, most do not check the remedies in a reference text. Age and training were negatively correlated in a statistically significant manner with (1) the likelihood of a physician prescribing alternative medicines; (2) checking the side effects and drug interactions of over the counter and prescription medications in a reference text; (3) asking patients specifically about alternative medicines; and (4) checking the side effects and drug interactions of alternative remedies in a reference text. In a question matching ten herbs and side-effects, the highest score was six out of ten correct and the average number correct was 1.32 with a standard deviation of 1.39. Clearly, physicians may be aware of different forms of alternative medicines. However, physicians are still not treating herbs in the same manner as other types of medications. There is no doubt that patient care would be greatly enhanced if physicians educated themselves and stayed in touch with their patients' beliefs and health care behavior. PMID- 11478564 TI - Nurses' perceptions of complementary and alternative medical therapies. AB - The purpose of this study was to identify the perceptions of nurses toward the effectiveness and safety, as well as their recommendations for and personal use of complementary and alternative medical therapies. A, random sample of 1000 nurses throughout the United States were surveyed using a three-wave mailing. About half of the respondents perceived there was conclusive evidence or preponderance of evidence that five therapies were effective: biofeedback, chiropractic, meditation/relaxation, multi-vitamins, and massage therapy. The same amount of nurses also perceived five therapies as definitely safe: hypnotherapy, chiropractic, acupressure, acupuncture, and healing touch. However, the nurses were most likely to recommend (regularly or periodically) four therapies: multivitamins, massage, meditation/relaxation, and pastoral/spiritual counseling. The vast majority (79%) of nurses perceived their professional preparation in this area to be fair or poor. PMID- 11478565 TI - Pneumococcal immunizations at flu clinics: the impact of community-wide outreach. AB - This study examined the effectiveness of a community-wide outreach campaign to promote the use of pneumococcal vaccine at public flu immunization clinics, and assessed whether this intervention was more effective than simply making pneumococcal vaccination available at such clinics. In 1997, a community-wide outreach campaign promoting pneumococcal and influenza immunizations was launched in a 17 zip code area of Dutchess County, NY. The campaign was aimed at 7,961 Medicare beneficiaries urging them to obtain pneumococcal immunization from local flu clinics. Medicare reimbursement data were used to assess the countywide pneumococcal vaccination rate, and to analyze differences between rates for beneficiaries in the target area and elsewhere in the county. Between 1996 and 1997 there was a 94% increase in pneumococcal vaccination billed to Medicare beneficiaries in Dutchess County. The 1997 annual rate of pneumococcal immunization in the target area reached 16.3% versus 12.2% elsewhere in the county (p < 0.001), with an increase over the previous year of 8.7% and 5.6%, respectively. Nearly all of the increase is accounted for by pneumococcal vaccination delivered at flu clinics. It is possible to significantly increase the use of pneumococcal immunization by linking its delivery to community-based flu clinics and by developing local outreach strategies. The outreach campaign has a significant additive effect over simply making PPV available at flu shot clinics. Additional community-wide outreach can further improve pneumococcal immunization utilization rates. PMID- 11478566 TI - Retention of low income mothers in a parenting intervention study. AB - Women with inadequate prenatal care were recruited to a multi-component parenting intervention study. Because it was anticipated that this high-risk population might present challenges to retention, a variety of strategies were employed to maintain their participation in the study. This report reviews the results of these retention efforts and compares the population that completed the study versus those that terminated prior to study completion. Two hundred and eighty six women were randomized to an intervention or control group. Careful tracking of the mothers, offering incentives for completing various study activities and providing a culturally competent staff were among the strategies employed to maintain participation. Comparison was made of those mothers terminating before study completion versus those retained, and of those terminating early in the study period versus later. Despite retention efforts, attrition at a level of 41% occurred. A few characteristics of mothers terminating early from the study were significant including older maternal age, a larger number of children, and incidence of no prenatal care. Despite comprehensive tracking procedures, some mothers were lost to follow up after change of residence. Other reasons for attrition included child outplacement and refusal of services or data collection procedures. PMID- 11478567 TI - A snapshot of homeless youth in Seattle: their characteristics, behaviors and beliefs about HIV protective strategies. AB - The purpose of this study was to determine how initial HIV prevention efforts for homeless youth were received and to determine areas where homeless youth's beliefs and behaviors continue to put them at risk for HIV infection. Interviews were conducted with 289 Seattle homeless youth. Youth reported using condoms with casual partners during vaginal and anal sex and with clients during oral, anal and vaginal sex. Condoms are often not used during vaginal sex with main partners or during oral sex with casual or main partners. Knowledge of HIV protective strategies differed according to youth's behavioral characteristics with heterosexual youth having the weakest knowledge of HIV protective strategies especially compared with young men who have sex with men. There is room for improvement in youth's knowledge and beliefs about HIV. PMID- 11478568 TI - Predicting presidential performance in the United States: equation replication on recent survey results. AB - For more than 2 decades, researchers have tried to identify the variables that predict the overall performance of U.S. presidents. In 1986, there emerged a 6 variable prediction equation (D. K. Simonton, 1986c, 1987b) that has been replicated repeatedly. The predictors are years in office, war years, scandal, assassination, heroism in war, and intellectual brilliance. The author again replicated the equation on recent rankings of all presidents from George Washington through William Jefferson Clinton according to a survey of 719 experts (W. R. Ridings, Jr., & S. B. McIver, 1997). The original 6-variable equation successfully predicted both the overall rankings as well as the 5 core components of the rankings (leadership qualities, accomplishment, political skill, appointments, character and integrity). The predictive value of the equation was illustrated for the presidencies of Ronald W. Reagan, George H. W. Bush, and Clinton. PMID- 11478569 TI - Relation of ethnicity and age to women's responses to personal experiences of sexist discrimination in the United States. AB - The authors explored self-reported experiences of interpersonal sexist discrimination and responses to such incidents in a heterogeneous sample of 262 women in the northeastern United States. They divided the sample into 2 categories for age (< 30 years, > 30 years) and for ethnicity (women of color, European American women). Across categories of age and ethnicity, the participants (a) commonly experienced sexist discrimination and (b) viewed men as the primary perpetrators of the discrimination. Although the women's reports of sexist experiences were similar for the most part, the authors found significant age differences in the frequency of categories of sexist events and in specific events, as well as in some general responses to such incidents. Ethnicity, operationalized here as women of color and European American women, did not have a significant influence. PMID- 11478570 TI - Intergroup differentiation: stereotyping as a function of status hierarchy. AB - The authors explored how negative intergroup comparisons affect intergroup differentiation. More specifically, they tested the prediction that the in group's negative intergroup comparisons with a high-status group would result in more negative stereotyping of a lower status out-group. The authors elicited stereotypes of a lower status university in 2 conditions. In the 1st, the participants judged only the middle-ranking university in-group and a lower status university. In the 2nd, those judgments followed comparison with a higher status university. In the 2nd condition, there was an increased differentiation between the in-group and the lower status out-group because of the more negative stereotyping of the lower status out-group. This evidence of intergroup differentiation was found only on the dimension judged most important and along which the in-group was negatively compared with the higher status group. PMID- 11478571 TI - Attitudes toward parenting strategies, potential for child abuse, and parental satisfaction of ethnically diverse low-income U.S. mothers. AB - Among a sample of 176 low-income mothers from 3 ethnic groups in the United States, the authors investigated ethnic differences in attitudes toward preferred parenting strategies, or styles; ethnic differences in the potential for child abuse; and the relationship between parenting strategies, the potential for child abuse, and parental satisfaction. They distributed the Maternal Reactions to Child's Deviant Behavior subscale (K. M. Rickard, W. Graziano, & R. Forehand, 1984), a shortened version of the Child Abuse Potential Inventory (CAPI; J. S. Milner & R. C. Wimberley, 1979), and a Parental Satisfaction Scale (N. P Medora, S. M. Wilson, & J. Larson, 1996) to the participants. The results indicated no significant ethnic differences in preferred parenting styles. Mothers from all 3 ethnic groups ranked praise and reasoning as the 1st and 2nd preferred parenting strategies. There were no ethnic differences in the perceived potential for child abuse. Parental satisfaction was negatively related to 2 of the CAPI subscales- Loneliness and Problems. The parenting strategy reasoning was positively correlated with parental satisfaction. PMID- 11478572 TI - Hindsight bias demonstrated in the prediction of a sporting event. AB - The authors examined hindsight bias in the context of a sporting event (Super Bowl XXXIII) with individuals who had previous knowledge of the hindsight bias-42 students who were currently enrolled in psychology classes that had previously covered hindsight bias. To control for extraneous variables, the authors analyzed how often the participants watched football, their gender, and whom they predicted to win; they found no differences between how often the participants watched football and whom they predicted to win. The hindsight bias existed for the prediction of the outcome for the Super Bowl, and the participants with previous knowledge were not immune to its effect. Results are discussed in relation to attribution theory, the reconstructionist view, and response bias view. PMID- 11478573 TI - Motivation for conflict among Chinese university students: effects of others' expertise and one's own confidence on engaging in conflict. AB - Researchers (A. C. Amason, 1996; D. H. Gruenfeld, 1995; K. A. Jehn, 1995, 1997; M. A. Rahim, 1989; M. A. Rahim & A. A. Blum, 1994; D. M. Schweiger, W. R. Sandberg, & P. L. Rechner, 1989; P. E. Tetlock, D. Armor, & R. S. Peterson, 1994) have documented the value of conflictual discussions for solving problems, but few have explored the conditions under which people are motivated to engage in controversy (K. A. Jehn, C. Chadwick, & S. M. B. Thatcher, 1997). Some (M. Van Berklom & D. Tjosvold, 1981) have hypothesized that high expertise and a competitive social context arouse concerns about defending one's position and challenging the opposing one. In the present study, Chinese university students in Hong Kong who expected to disagree with an expert, compared with those who did not expect to disagree with an expert, had less confidence, felt less knowledgeable about their position, and selected an agreeable discussant. Consistent with the idea of maintaining distance from those in power, the participants were reluctant to disagree directly with someone with greater expertise. PMID- 11478574 TI - The effects of personal need for structure and occupational identity in the role stress process. AB - The authors formed and tested a model of the antecedents, consequences, and possible moderators or mediators of the role ambiguity construct in a new occupational group of nurses (N = 1,240) in Finland. They hypothesized that (a) job characteristics (control and feedback) would be associated with role ambiguity and (b) this association, in turn, would be moderated by personal need for structure. The authors hypothesized that role ambiguity would affect occupational strain partially by the strength of occupational identity. They tested these models by using hierarchical regression analysis and structural equation modeling (LISREL 8; K. G. Joreskog & D. Sorbom, 1993). Both hypotheses were supported. PMID- 11478575 TI - The moderating effect of teamwork perceptions on the organizational politics--job satisfaction relationship. AB - By using regression analyses on data from 355 full-time employees of a customer service organization in the eastern United States, the authors tested the hypothesis that perceptions of organizational politics are more strongly related to job dissatisfaction among individuals who perceive low levels of teamwork importance than among those who perceive high levels of teamwork importance. Hierarchical moderated regression analysis of the data revealed that the moderating effect of teamwork importance was most relevant at average-to-high levels of perceived politics. That finding supports the assertion that one way to address the negative impact of organizational politics is to try to ensure that employees value teamwork. PMID- 11478576 TI - Relations of self-esteem concerns, group identification, and self-stereotyping to in-group favoritism. AB - In 2 studies, the author examined the effect of collective self-esteem (CSE; J. Crocker & R. Luhtanen, 1993) on people's willingness to display in-group favoritism. To test that self-esteem hypothesis, he measured public CSE, rather than private CSE, because the former parallels a threat to social identity, a state believed to motivate in-group favoritism. Furthermore, the author explored whether group identification and self-stereotyping moderated the effect of public CSE on in-group favoritism. The participants were 92 British and Dutch university employees. As expected, participants high in public CSE displayed more in-group favoritism than did those low in public CSE. Moreover, group identification and self-stereotyping appeared to moderate the effect of CSE. PMID- 11478577 TI - Self-esteem and in-group bias among members of a religious social category. AB - In a sample of New Zealand university students, the author extended earlier research into the relationship between self-esteem and intergroup discrimination. He found no support for the hypothesis that social-category members (i.e., Christians) experience an elevation in the domain of self-esteem (i.e., religious self-esteem) judged as more relevant to the in-group after evaluations favoring the in-group. Regardless of whether the evaluation targets behaved positively or negatively, the respondents in the experimental condition evaluated in-group (Christian) targets more highly than out-group (Atheist) targets. After evaluations favoring the in-group, the respondents did not experience an elevation of religious self-esteem, global self-esteem, or mathematical self esteem (judged as less relevant to the in-group). PMID- 11478578 TI - The influence of status on pedestrians' failure to observe a road-safety rule. PMID- 11478579 TI - The failure of HAART to cure the HIV-1/AIDS complex. Suggestions to add integrase inhibitors as complementary virostatics, and to replace their continuous long combination applications by short sequences differing by drug rotations. AB - While the intensive virostatic combinations applied according to the conventional models (such as HAART), based only on the attacks of two HIV-1 targets, retrotranscriptase and protease, and applied in a long and continuous fashion, a) are notably toxic, b) do not correct completely the abnormal immunologic parameters, and c) are followed by particularly severe and poorly sensitive relapses in case of discontinuation, we propose to the 'AIDS treatment headquarters' to include in their failing strategy the two original features which we have included in the treatment of a cohort of a dozen patients, treatment applied at all but one AIDS stage. We attack one more HIV-1 target than the conventional protocols do, by adding inhibitors of integrase; we apply the combinations of virostatics, comprising inhibitors of the three targets, in short sequences (of 3 weeks), between which the analogues are changed inside each series. The first patient of the cohort started his treatment 8.5 years ago, and the entries of the others into it have been at random and not randomized. All patients are alive today and in excellent condition. PMID- 11478580 TI - Current review and clinical management of patients with primary HIV-1 infection: limits and perspectives. AB - Acute primary HIV-1 infection (PHI) remains underestimated or misdiagnosed in clinical practice. Meanwhile, it has been demonstrated that early therapeutic interventions with highly active antiretroviral therapy (HAART) may delay disease progression and possibly preserve and expand the most effective immune effector T cells against HIV together with the humoral immune responses. Since long-life HAART is an unachievable goal due to long-term toxicity and risk of occurrence of resistant strains due to a decreased compliance or other still undefined host factors, preliminary data of programmed treatment interruption in patients treated for PHI suggest that a significant number (30-50%) could benefit from long periods off therapy. However, in more than half of them, the viral load will rebound, justifying that treatment be reinitiated. In order to reduce this proportion, new options are currently being investigated, including adjunctive immune therapy to HAART such as cytokines or vaccines, which could tackle the viral rebounds by increasing HIV-specific cellular responses. An update on the management of patients with PHI is reviewed and the limits of the current standard of care are discussed. PMID- 11478581 TI - Additive effect of tunicamycin and dextran sulfate on the binding of monoclonal antibody to the V2 domain of the envelope glycoprotein 120 of human immunodeficiency virus type 1. AB - Using flow cytometry or immunoprecipitation analysis in cells chronically infected with HIV-1 IIIB Supt-1, we noticed an additive effect of tunicamycin and low molecular weight dextran (LMDS) on the binding of the G3-4 monoclonal antibody to monomeric and oligomeric forms of glycoprotein 120 (gp120). The inhibition of glycosylation by tunicamycin reduced the number of monomeric and oligomeric forms of gp120. The inhibition of the binding of the G3-4 antibody to monomeric and oligomeric forms of gp120 was more pronounced in the presence of LMDS. We also found that the G3-4 antibody can not recognise the nascent polypeptide chain of the envelope glycoprotein. PMID- 11478582 TI - Antibody responses to HIV-1 antigens are higher in HIV-1(+) intravenous drug users than in HIV-1(+) homosexuals. AB - Immune responses to HIV-1 infection of 42 HIV-1-positive asymptomatic intravenous drug users (IVDUs) were compared with those of 135 HIV-1-infected asymptomatic homosexual men in the present study. Twenty-five HIV-1(-) individuals served as normal controls. The comparison included antibody responses to five computer predicted epitopes of HIV-1 p17, and viral proteins gp120 and p24 as well as p17. Major immunophenotypes were also investigated. Results showed that antibody responses to the five epitopes were significantly higher in the IVDUs. A larger proportion of the IVDUs, with respect to that of homosexuals, showed positive antibody responses to p24 and p17, respectively. However, the antibody response to gp120 was similar between the two cohorts. Immunophenotyping showed that HIV 1(+) homosexuals had higher profiles in most of the major subsets than did the IVDUs, especially in the total count of lymphocytes, absolute numbers of CD3+ cells and CD8+ cells. It appeared that the HIV-1(+) IVDU cohort had higher antibody responses to most of the viral antigens, but had lower levels of lymphocyte subsets in comparison with HIV(+) homosexuals. PMID- 11478583 TI - Induction in mice of anti-Tat mucosal immunity by the intranasal and oral routes. AB - Anti-Tat vaccination experiments were carried out in mice with a view to inducing systemic in addition to mucosal immunity. For this, three types of immunizing preparations were tested, which consisted of Tat toxoid embedded in either an adjuvant oily structure (IMS), or nanoparticles of chitosan, or microparticles of polylactide-co-glycolide (PLG). Administered by either the intranasal or oral route all preparations triggered anti-Tat IgG and IgA antibodies. Sera from mice immunized with either of these preparations could also inhibit significantly the Tat transactivating activity. These results open up a new avenue to the development of an effective anti-AIDS protective vaccine. PMID- 11478584 TI - Predictors of protease inhibitor-associated adverse events. AB - Risk factors in the development of adverse reactions in HIV-1-infected patients treated with highly active antiretroviral therapy (HAART) containing protease inhibitors are poorly understood. To identify predictors of protease inhibitor associated adverse events, we are conducting a prospective, cohort, multicenter study on HIV-positive patients starting treatment with at least one protease inhibitor. Rate ratios (RR) of adverse events were calculated, and logistic regression was used to adjust simultaneously for the potentially confounding effects of selected variables, according to the Cox model. A total of 1477 patients have been enrolled up to April 2000, having an average age of 37.1 years (SD +/- 8.1); 1066 (72.2%) were male. Where risk factors for HIV infection are concerned, the distribution was as follows: 48.1% intravenous drug users, 31.6% heterosexual contacts, 16.2% homosexual males and 0.7% blood transfusion. Average CD4+ lymphocyte count at enrollment was 265 cells/mmc (SD +/- 201). Average follow-up time is equal to 17.8 months (range 1-32). The risk of developing adverse reactions is significantly increased in female patients, older patients, hemophiliac subjects and in subjects with hepatitis. Patients treated with ritonavir, the association ritonavir-saquinavir HGC, stavudine and efavirenz have significantly increased incidence of adverse reactions in PI-containing regimens; conversely, saquinavir HGC, zidovudine and lamivudine use was associated with a lower risk of developing adverse reactions. PMID- 11478585 TI - Iron: deficiencies and requirements. AB - A report from the World Health Organization estimates that 46% of the world's 5- to 14-year-old children are anemic. In addition, 48% of the world's pregnant women are anemic. A majority of these cases of anemia are due to iron deficiency. Our aim here is to review the latest data on iron regulatory mechanisms, iron sources and requirements. Human and animal studies have shown that amino acids and peptides influence iron absorption from the intestinal lumen. Inter-organ transport and uptake of nonheme iron is largely performed by the complex transferring-transferring receptor system. Moreover, the discovery of cytoplasmic iron regulatory proteins (IRPs) has provided a molecular framework from which we understand the coordination of cellular iron homeostasis in mammals. IRPs and the iron responsive elements (IREs) to which they bind allow mammals to make use of the essential properties of iron while reducing its potentially toxic effect. Physiologic iron requirements are three times higher in pregnancy than they are in menstruating women (approximately 1200 mg must be acquired from the body's iron store or from the diet by the end of pregnancy). The administration of iron supplements weekly instead of daily in humans has been proposed and is being actively investigated as a viable means of controlling iron deficiency in populations, including pregnant women. PMID- 11478586 TI - Iron metabolism, free radicals, and oxidative injury. AB - Iron has the capacity to accept and donate electrons readily. This capability makes it physiologically essential, as a useful component of cytochromes and oxygen-binding molecules. However, iron is also biochemically dangerous; it can damage tissues by catalyzing the conversion of hydrogen peroxide to free-radical ions that attack cellular membranes, protein and DNA. This threat is reduced in the healthy state where, because of the fine iron metabolism regulation, there is never appreciable concentration of 'free iron'. Under pathological conditions, iron metabolism and superoxide metabolism are clearly interactive. Each can exacerbate the toxicity of the other. Iron overload may amplify the damaging effects of superoxide overproduction in a very broad spectrum of inflammatory, both acute and chronic, conditions. Furthermore, chronic oxidative stress may modulate iron uptake and storage, leading to a self-sustained and ever-increasing spiral of cytotoxic and mutagenic events. The iron chelator deferroxamine is able to chelate 'free iron' even inside the cell. Its regular clinical use is to promote the excretion of an iron overload, when phlebotomy is harmful, and the dosage varies between 2-10 g/d. In conditions where deferroxamine is used to prevent the iron-driven oxygen toxicity, i.e., acute or chronic inflammatory diseases with oxidative stress, the dosage can be extremely reduced and the addition of antioxidants could be useful. PMID- 11478587 TI - A comparison of no-flow and low-flow ischemia in the rat heart: an energetic study. AB - Heat production under no-flow ischemia (ISCH) and under hypoperfusion (HYP) conditions was measured in single isovolumetric contractions of perfused rat ventricles at 25 degrees C. Resting heat production (Hr) and resting pressure decreased when the perfusion rate was reduced from 6 to 1.5 mL min(-1) or lower flows (HYP) and by ISCH. Maximal developed pressure (P) decreased to 29% and 20% of control by HYP at 0.8 mL min(-1) and ISCH, respectively. The tension independent heat (TIH) fraction attributed to Ca2+-binding, measured during single contractions, decreased under HYP with an increase in the ratio between the maximum relaxation rate and P (-P/P ratio). The TIH fractions (attributed to Ca2+ binding and Ca2+ removal processes) decreased under ISCH. The long duration TIH fraction associated with Ca2+-dependent mitochondrial activity disappeared at flow rates of 1.5 mL min(-1) or lower. The ratio between the tension-dependent energy release and P was decreased by ISCH but not by HYP, indicating that under ISCH there was an improvement in contractile economy, but this was not modified by HYP. Overall, the results indicate that no-flow and low-flow ischemias are energetically different models. While the contractile failure under HYP seems to be related to a decrease in myofilament Ca2+ sensitivity, under ISCH it appears to be related to decreased cytosolic Ca2+ availability combined with a more noticeable effect on a fraction of energy that has been attributed to mitochondrial activity. Furthermore, mechanical and energetic responses of both models (i.e., ISCH and HYP) found in the present work were not the same as those previously observed in severe hypoxia so that all these models should not be used indistinctly. PMID- 11478588 TI - Caffeine ingestion elevates plasma insulin response in humans during an oral glucose tolerance test. AB - We tested the hypothesis that caffeine ingestion results in an exaggerated response in blood glucose and (or) insulin during an oral glucose tolerance test (OGTT). Young, fit adult males (n = 18) underwent 2 OGTT. The subjects ingested caffeine (5 mg/kg) or placebo (double blind) and 1 h later ingested 75 g of dextrose. There were no differences between the fasted levels of serum insulin, C peptide, blood glucose, or lactate and there were no differences within or between trials in these measures prior to the OGTT. Following the OGTT, all of these parameters increased (P < or = 0.05) for the duration of the OGTT. Caffeine ingestion resulted in an increase (P < or = 0.05) in serum fatty acids, glycerol, and plasma epinephrine prior to the OGTT. During the OGTT, these parameters decreased to match those of the placebo trial. In the caffeine trial the serum insulin and C peptide concentrations were significantly greater (P < or = 0.001) than for placebo for the last 90 min of the OGTT and the area under the curve (AUC) for both measures were 60 and 37% greater (P < or = 0.001), respectively. This prolonged, increased elevation in insulin did not result in a lower blood glucose level; in fact, the AUC for blood glucose was 24% greater (P = 0.20) in the caffeine treatment group. The data support our hypothesis that caffeine ingestion results in a greater increase in insulin concentration during an OGTT. This, together with a trend towards a greater rather than a more modest response in blood glucose, suggests that caffeine ingestion may have resulted in insulin resistance. PMID- 11478589 TI - Hemodynamic effects of Salvia miltiorrhiza on cirrhotic rats. AB - Salvia miltiorrhiza (Sm) administration has been shown to reduce hepatic fibrosis in rats. We investigated the hemodynamic effects of Sm on bile duct ligated (BDL) rats. Hemodynamic, histological, and vascular contractile studies were conducted in rats 4 weeks after bile duct ligation. An aqueous extract of Sm (0.2 g twice per day) or vehicle was administered for 4 weeks to BDL rats. Sm treatment in BDL rats significantly reduced histological grades of fibrosis and ameliorated the portal hypertensive state (including portal venous pressure, superior mesenteric artery blood flow, cardiac index, and total peripheral resistance) as compared with vehicle treatment. Moreover, Sm treatment enhanced the vascular sensitivity of mesenteric arteries to phenylephrine in BDL rats. Sm treatment had no effect on plasma biochemical profiles of either BDL or normal rats. Our results suggest that 4-week Sm treatment ameliorates the portal hypertensive state in BDL rats. PMID- 11478590 TI - Thiocyanate as a probe of the cystic fibrosis transmembrane conductance regulator chloride channel pore. AB - Immediately following exposure to thiocyanate (SCN-)-containing solutions, the cystic fibrosis conductance regulator Cl- channel exhibits high unitary SCN conductance and anomalous mole fraction behaviour, suggesting the presence of multiple anion binding sites within the channel pore. However, under steady-state conditions SCN-conductance is very low. Here I show, using patch clamp recording from CFTR-transfected mammalian cell lines, that under steady-state conditions neither SCN- conductance nor SCN- permeability show anomalous mole fraction behaviour. Instead, SCN conductance, permeability, and block of Cl- permeation can all be reproduced by a rate theory model that assumes only a single intrapore anion binding site. These results suggest that under steady-state conditions the interaction between SCN- and the CFTR channel pore can be understood by a simple model whereby SCN- ions enter the pore more easily than Cl-, and bind within the pore more tightly than Cl-. The implications of these findings for investigating and understanding the mechanism of anion permeation are discussed. PMID- 11478591 TI - Serotonin reuptake inhibitors fluoxetine and citalopram relax intestinal smooth muscle. AB - Selective serotonin reuptake inhibitor antidepressants (SSRIs) exert depressant effects on cardiac myocytes and vascular smooth muscle cells by inhibiting Ca2+ channels. We hypothesized that the SSRIs fluoxetine and citalopram affect the contractile activity of intestinal smooth muscle by interfering with Ca2+ entry and (or) signaling pathways. The effects of fluoxetine and citalopram on contractions of guinea-pig ileum longitudinal muscle-myenteric plexus preparations (LMMP) were compared with the effects of the voltage-operated Ca2+ channel inhibitors nifedipine and diltiazem. In a concentration-dependent manner, nifedipine, diltiazem, fluoxetine, and citalopram elicited relaxation of LMMPs contracted by electrical field stimulation (EC50 values of 4 x 10(-7) M, 1.4 x 10(-6) M, 1.4 x 10(-5), and 6.8 x 10(-6) M, respectively). Nifedipine, diltiazem, fluoxetine, and citalopram also relaxed LMMPs contracted with a depolarizing concentration of KCl (48 mM; EC50 values of 1.8 x 10(-8) M, 1.4 x 10(-7) M, 3.7 x 10(-6) M, and 6.3 x 10(-6), respectively), a response that could be reversed by increasing the extracellular Ca2+ concentration (2.5-30 mM). These data suggest that fluoxetine and citalopram elicit relaxation of intestinal smooth muscle, likely by inhibiting Ca2+ channel(s). This effect may be of clinical importance. PMID- 11478592 TI - Arotinolol is a weak partial agonist on beta 3-adrenergic receptors in brown adipocytes. AB - Arotinolol, a clinically used alpha/beta-adrenergic blocker, has been demonstrated to be an anti-obesity agent. The anti-obesity effect of arotinolol was suggested to be the result of direct activation of thermogenesis in brown-fat cells. We tested the ability of arotinolol to stimulate thermogenesis (oxygen consumption) in isolated brown-fat cells and in intact animals. Arotinolol stimulated thermogenesis in brown-fat cells isolated from mouse and hamster. A relatively low sensitivity to the beta-adrenergic antagonist propranolol (pK(B) approximately 6) indicated that arotinolol interacted with the beta3-adrenergic receptor. On the beta3-receptor, arotinolol was a very weak (EC50 approximately 20 microM) and only partial (approximately 50%) agonist, but arotinolol also demonstrated the properties of being a beta3-receptor antagonist with a pK(B) of 5.7. In intact animals, only the antagonistic action of arotinolol could be observed. Because arotinolol is only a very weak and partial agonist on the beta3 receptors, direct stimulation of thermogenesis in brown adipose tissue is unlikely to be sufficient to cause significant weight loss. It may be necessary to invoke additional pathways to explain the anti-obesity effects of chronic treatment with arotinolol. PMID- 11478593 TI - Intravenous lipid composition affects hypoxic pulmonary vasoconstriction in the newborn piglet. AB - To examine the effects of altering the fatty acid (FA) composition of intravenous (IV) lipid emulsions on pulmonary vascular resistance (PVR) and thromboxane production, we studied three groups of newborn piglets after three days of either sow's milk (milk), or total parenteral nutrition (TPN) with either iv soy bean oil (SBO, 52% n-6 and 8% n-3 FA) or fish oil (FO, 5% n-6 and 51% n-3 FA) emulsions. At baseline, and during hypoxia at 20 min and 2 h, cardiac output (Q) was measured, PVR calculated and plasma levels of a prostacyclin metabolite (6 keto-PgF1alpha) and thromboxane B2 (TxB2) were measured. Fatty acid composition of the lung phospholipids was analyzed. There was an exaggerated increase in PVR and decrease in Q during prolonged hypoxia in the TPN-SBO group as compared with the other two groups. There was no difference in PVR and Q between the milk and TPN-FO groups. FA of lung phospholipids reflected the high dietary level of long chain n-3 FA in the TPN-FO group. However, no differences in plasma levels of 6 keto-PgF1alpha or TxB2 were found. Intravenous emulsions made from SBO reduced cardiac output and increased pulmonary vascular resistance in the hypoxic newborn piglet, whereas iv FO emulsions did not. When subjects with pulmonary hypertension are receiving TPN iv SBO may be detrimental; iv FO may be beneficial, giving similar responses as in a milk-fed subject. PMID- 11478594 TI - Effect of chronic and progressive aortic constriction on renal function and structure in rats. AB - The purpose of this study was to evaluate the functional and structural renal damage observed in aortic-constricted hypertensive rats and to identify their possible relationship with transforming growth factor beta (TGF-beta) expression. Progressive renovascular hypertension was induced by progressive aortic constriction between the two renal arteries. Three months after constriction, the glomerular filtration rate (GFR), effective renal blood flow (ERBF), perfusion pressure (PP), urinary protein excretion (UPE) and urinary electrolyte excretion (U(Na)V and U(K)V) in the kidney above (right kidney, RK) and below the ligature (left kidney, LK) were measured. The cross-sectional corpuscular, capillary tuft and mesangial matrix area and tubulo-interstitial fibrosis were measured in tissue sections stained with Syrius Red using a computer-assisted image analysis system. TGF-beta was detected by immunohistochemistry. The functional parameters were similar in the two kidneys of aortic-constricted hypertensive rats (GFR-RK, 1.33+/-0.08 vs. LK, 1.18+/-0.08 mL/min; ERBF-RK, 9.23+/-1.32 vs. LK, 8.18+/-0.91 mL/min; RVR-RK, 28.3+/-3.9 vs. LK, 21.7+/-3.2 mmHg x min/mL). The RK was subject to a higher PP than the LK (176+/-7 vs. 128+/-5 mmHg, P < 0.05). UPE, U(Na)V, and U(K)V were greater in the RK than in the LK (UPE-RK, 512+/-61 vs. LK, 361+/-38 microg/30 min, P < 0.05; U(Na)V-RK, 0.056+/-0.012 vs. LK, 0.022+/-0.006 mEq/30 min, P < 0.05; UKV-RK, 0.042+/-0.006 vs. LK, 0.029+/-0.003 mEq/30 min, P < 0.05). Morphometric analysis revealed that the RK capillary tuft area and mesangial matrix area were higher than those in the LK. The LK had a higher degree of interstitial fibrosis than the RK. No significant differences in TGF-beta immunostaining were observed between the RK and the LK. In conclusion, the RK (subjected to hypertension) of aortic-constricted hypertensive animals developed glomerular fibrosis, only in the outer glomeruli whereas the LK developed mild interstitial fibrosis. Neither glomerular nor interstitial fibrosis seem to be responsible for the proteinuria observed in both kidneys. PMID- 11478595 TI - Diversity of K+ channels in circular smooth muscle of opossum lower esophageal sphincter. AB - We previously demonstrated that a balance of K+ and Ca2+-activated Cl- channel activity maintained the basal tone of circular smooth muscle of opossum lower esophageal sphincter (LES). In the current studies, the contribution of major K+ channels to the LES basal tone was investigated in circular smooth muscle of opossum LES in vitro. K+ channel activity was recorded in dispersed single cells at room temperature using patch-clamp recordings. Whole-cell patch-clamp recordings displayed an outward current beginning to activate at -60 mV by step test pulses lasting 400 ms (-120 mV to +100 mV) with increments of 20 mV from holding potential of -80 mV ([K+]I = 150 mM, [K+]o = 2.5 mM). However, no inward rectification was observed. The outward current peaked within 50 ms and showed little or no inactivation. It was significantly decreased by bath application of nifedipine, tetraethylammonium (TEA), 4-aminopyridine (4-AP), and iberiotoxin (IBTN). Further combination of TEA with 4-AP, nifedipine with 4-AP, and IBTN with TEA, or vice versa, blocked more than 90% of the outward current. Ca2+-sensitive single channels were recorded at asymetrical K+ gradients in cell-attached patch clamp configurations (100.8+/-3.2 pS, n = 8). Open probability of the single channels recorded in inside-out patch-clamp configurations were greatly decreased by bath application of IBTN (100 nM) (Vh = -14.4+/-4.8 mV in control vs. 27.3+/ 0.1 mV, n = 3, P < 0.05). These data suggest that large conductance Ca2+ activated K+ and delayed rectifier K+ channels contribute to the membrane potential, and thereby regulate the basal tone of opossum LES circular smooth muscle. PMID- 11478596 TI - Effect of tetramethyl pyrazine on L-type calcium channel in rat ventricular myocytes. AB - To elucidate possible ionic mechanisms of antimyocardial ischemia and antiarrythmia of tetramethyl pyrazine (TP), we studied L-type Ca2+ currents (I(Ca.L)) in adult rat ventricular myocytes using the whole-cell patch-clamp technique. The results showed: (i) under physiological conditions, 0.25 mmol/L TP decreased amplitude of I(Ca.L) to 60.6% and this inhibition was increased with increasing concentration of TP. ID50 was 0.20 mmol/L. (ii) The Ca2+-antagonistic effect of TP was voltage-dependent. A marked negative shift of the steady-state inactivation curve was observed with long (10 s) conditioning prepulses, but not with short (350 ms) ones. (iii) The time course of inhibition during TP treatment was increased with an increase in drug concentration, and recovery from TP induced inactivation of I(Ca.L) was slower than in control cases. (iv) Tonic block and use-dependent block with TP treatment, which was induced by increasing the frequency of stimulation, occurred. We suggest that TP inhibits the I(Ca.L) mainly by binding to inactivated Ca2+ channels. The high affinity of TP for the inactivated state of I(Ca.L) may play an important role in developing therapies for pathological conditions. PMID- 11478597 TI - Metabolism of angiotensin I by guinea pig aqueous humor. AB - We investigated the degradation of angiotensin I (Ang I) by guinea pig aqueous humor at physiological pH (pH 7.4) and assessed the activity of responsible enzymes using various enzyme inhibitors. The aqueous humor was incubated with Ang I in the presence or absence of an enzyme inhibitor at 37 degrees C for the appropriate time period. The resulting peptides were analyzed by a Beckman HPLC system with a Waters microBondapak C18 analytical column using a 30-min increasing linear gradient of 10 to 40% acetonitrile containing 0.05% trifluoroacetic acid (TFA) and H2O containing 0.05% TFA at a flow rate of 1 mL/min. Detection was done by absorbance at 214 nm. Angiotensin II (Ang II) was a major product (39.3+/-4.10 nmol x h(-1) mL(-1), n = 5) of Ang I hydrolysis. Traces of angiotensin 1-9, angiotensin IV, and angiotensin 1-7 were also produced. Chymostatin (0.05 mmol/L), EDTA (1 mmol/L), enalaprilat (0.1 mmol/L), and ebelacton B (0.01 mmol/L) inhibited generation of Ang II from Ang I by guinea pig aqueous humor by 89+/-4.6, 56+/-7.6, 33+/-5.1, 20+/-6.5%, respectively. Our findings indicate that guinea pig aqueous humor contains several enzymes that can form Ang II. The chymostatin-sensitive type of enzyme was the most active one found in guinea pig aqueous humor. Angiotensin I converting enzyme, carboxypeptidase A, and deamidase may also contribute to angiotensin II formation in guinea pig ocular fluid. PMID- 11478598 TI - Complex vertebral malformation in holstein calves. AB - A recently observed lethal congenital defect of purebred Holstein calves is reported. Eighteen genetically related calves were necropsied. One calf had been aborted on gestation day 159, and the others were delivered between day 250 and day 285. Birth weights were reduced. The defect was characterized by shortening of the cervical and thoracic parts of the vertebral column due to multiple hemivertebrae, fused and misshaped vertebrae, and scoliosis. Symmetrical flexures of the carpal joints and the metacarpophalangeal joint in combination with a slight lateral rotation of the phalanges also were present. Similar low-grade arthrogryposis was present in the posterior limbs. Fifty percent of the calves had heart malformation. Other malformations occurred in a few calves. Complex vertebral malformation (CVM) is proposed as the designation for this defect. A genetic etiology is indicated because cases occurred following breeding between genetically related individuals. Two common ancestors were found; both were former elite sires of US Holstein origin. Because of the widespread international use of semen from sires occurring in the pedigrees of affected calves, CVM is expected to occur in several countries. PMID- 11478599 TI - Partial passive protection with two monoclonal antibodies and frequency of feeding of hyperimmune anti-transmissible gastroenteritis virus (TGEV) serum for protection of three-day-old piglets from a TGEV challenge infection. AB - Passive protection experiments were conducted to determine the frequency and amounts of hyperimmune antiserum needed to block a transmissible gastroenteritis virus (TGEV) challenge infection and to identify monoclonal antibodies that are partially protective against TGEV. Hyperimmune antiserum or monoclonal antibodies were added to milk at each feeding or at selected feedings when the amount of antiserum was reduced. Three-day-old piglets were challenged with virulent virus that had been preincubated with antiserum or monoclonal antibodies. The results indicated that supplementing antiserum every other day was not efficacious for protection. Supplementing even small quantities of hyperimmune antiserum (0.5 ml) at least once a day in most cases was sufficient for piglet survival but did not prevent morbidity. Increasing the amount (>2 ml) and providing antiserum 3 times/day completely blocked the TGEV challenge infection. Two monoclonal antibodies were discovered that also provided passive protection for baby pigs. One monoclonal antibody, 5G1, had a high neutralizing titer, and the other, 6C4, was more effective in neutralizing and binding to virulent TGEV than to attenuated TGEVs. Both of these monoclonal antibodies were partially effective as supplements in milk for passive protection. Furthermore, these monoclonal antibodies were useful for boosting the efficacy of TGEV-neutralizing colostrum, which by itself was ineffective. These results show that other antigenic sites, different from the 4-well characterized epitopes on the S glycoprotein of TGEV, also are important for passive protection. PMID- 11478600 TI - Application of a direct flow cytometric erythrocyte immunofluorescence assay in dogs with immune-mediated hemolytic anemia and comparison to the direct antiglobulin test. AB - A direct flow cytometric erythrocyte immunofluorescence assay (FC) was developed and compared with the direct antiglobulin test (DAT) for detection of erythrocyte bound immunoglobulin (IgG and IgM) and complement (C3) in dogs with immune mediated hemolytic anemia (IMHA). Tests were performed on erythrocytes from 13 healthy nonanemic dogs and from 13 anemic dogs with IMHA. The FC and DAT were negative for erythrocyte-bound immunoglobulin in all healthy dogs. The FC was negative for erythrocyte-bound C3 in 12 healthy dogs and positive in 1 healthy dog, and the DAT was negative for C3 in all healthy dogs. Of the 13 IMHA dogs tested for erythrocyte-bound IgG, 12 were positive using the FC and 7 were positive using the DAT. Sensitivity for the detection of erythrocyte-bound IgG in the 26 dogs was 92% for FC and 53% for DAT. Specificity for detection of erythrocyte bound IgG for FC and DAT was 100%. The addition of IgM and/ or C3 did not increase the sensitivity for FC or DAT. In this group of dogs, the FC provided a more rapid, cost-effective, sensitive, objective method to quantitate erythrocyte-bound immunoglobulin and/or complement compared with the currently used DAT. PMID- 11478601 TI - Early detection of maedi-visna (ovine progressive pneumonia) virus seroconversion in field sheep samples. AB - The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples (n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (K value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV infected sheep in field conditions and has potential for use in control and eradication programs. PMID- 11478602 TI - Multiplex polymerase chain reaction assay for identification of enterotoxigenic Escherichia coli strains. AB - A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein (uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non-E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates. PMID- 11478603 TI - Evaluation and use of a nested polymerase chain reaction assay in cats experimentally infected with Bartonella henselae genotype I and Bartonella henselae genotype II. AB - Cats have been shown to be infected with Bartonella henselae genotype I, B. henselae genotype II, and B. clarridgeiae. Feline bartonellosis infections and the strains involved in these infections are important in both veterinary and human medicine. Nucleic acid amplification methods such as polymerase chain reaction (PCR) are being used in both research and diagnostics as tools for understanding many infectious diseases. Bartonella bacteremia in cats is detected by blood culture; however, because of the limitations of culture (delayed turnaround time and sensitivity limits), PCR may be a more efficient method for identifying infected cats. Three distinct PCR assays that could differentiate among B. henselae genotype I, B. henselae genotype II. and B. clarridgeiae were developed and used to detect as few as 3.2 organisms. Fourteen cats experimentally infected with B. henselae genotype I and B. henselae genotype II were followed by bacterial culture and PCR through the course of infection, including periods of primary and relapsing bacteremia. The PCR assay was positive in 11 of the 14 cats for periods of 1-9 weeks after culture became negative. Of the 223 blood specimens that were culture negative, the PCR assay was positive in 38 (17%) of the specimens. Two of the 14 cats developed relapsing bacteremia. The 2 B. henselae genotypes were amplified in the cats and the bacteremic phase of these infections as determined by PCR lasted for a longer period than previously determined by culture. Using laboratory assays such as PCR to understand the strains involved in feline bartonellosis and the course of the infection is important in the understanding of these zoonotic agents. PMID- 11478604 TI - Correlation of ergovaline and lolitrem B levels in endophyte-infected perennial ryegrass (Lolium perenne). AB - The varieties of perennial ryegrass (Lolium perenne) infected with the endophytic fungus Neotiphodium lolii contain several classes of toxic alkaloids, including ergopeptide alkaloids and lolitrem alkaloids. Lolitrem B, a potent tremorgen, is generally considered to be the predominant alkaloid in endophyte-infected perennial ryegrass. Ergovaline, a vasoconstrictor normally associated with endophyte-infected tall fescue (Festuca arudinacea), is also present in endophyte infected perennial ryegrass. Clinical signs of animals ingesting endophyte infected perennial ryegrass are consistent with the presence of lolitrem B. However, clinical signs normally associated with ergovaline poisoning are not usually observed in animals ingesting endophyte-infected perennial ryegrass. A survey was conducted to quantitate both lolitrem B and ergovaline in 459 perennial ryegrass straw samples received at the Oregon State University College of Veterinary Medicine. Samples were analyzed for each alkaloid using separate high-performance liquid chromatography analyses. A strong positive correlation between the 2 alkaloids (r2 = 0.7335) was observed, especially in the samples containing <3,000 ppb (ng/g) lolitrem B. The threshold levels above which clinical signs typically occur are 2,000 ppb lolitrem B and 300-400 ppb ergovaline. All of the samples analyzed contained <425 ppb ergovaline. PMID- 11478605 TI - Immunoreactivity of A103, an antibody to Melan A, in canine steroid-producing tissues and their tumors. AB - The monoclonal antibody A103 to the melanocytic differentiation antigen Melan A stains human steroid-producing cells and their tumors. A total of 200 formalin fixed, paraffin-embedded canine normal tissues and hyperplastic and neoplastic lesions of the adrenal gland, testis, and ovary were immunohistochemically tested for Melan A with antibody A103. Leydig cell tumors (23/23, 100%), Sertoli cell tumors (14/15, 93%), and adrenocortical adenomas (12/13, 92%) were consistently positive. Adrenocortical carcinomas (23/35, 65%) and granulosa cell tumors (10/17, 59%) were less frequently positive. All pheochromocytomas, seminomas, and dysgerminomas were negative. The pattern of staining was cytoplasmic, but nuclear staining was also frequently seen in normal Leydig cells and their tumors. As in human tumors, immunohistochemistry for Melan A stains many canine steroid producing tumors and can be used to distinguish these tumors from those of nonstereidogenic cells. PMID- 11478606 TI - Complex vertebral malformation in a holstein calf: report of a case in the USA. AB - Complex vertebral malformation (CVM), a familial syndrome of Holstein calves, has been reported in aborted fetuses and in prematurely born, stillborn, and neonatal calves. Affected calves have anomalies in the vertebral column, including hemivertebrae, fused and misshapen vertebrae and ribs, scoliosis, and vertebral synostosis. Concurrent low body weight, symmetrical arthrogryposis, and cardiac anomalies have been documented in affected calves. The syndrome was identified and characterized in Holstein cattle in Denmark; however, a global distribution of this genetic disorder is likely based on identification of common ancestral sires widely used for artificial insemination. This is the first documented case of CVM in a Holstein calf in the USA. PMID- 11478607 TI - Nucleotide sequence and polymerase chain reaction/restriction fragment length polymorphism analyses of Aleutian disease virus in ferrets in Japan. AB - Two ferrets with spontaneous Aleutian disease (AD) were found in Japan. The diagnosis was verified by polymerase chain reaction (PCR) amplification of part of the capsid gene specific to AD virus (ADV). The nucleotide sequences (365 bp in length) of the amplified fragments from the 2 ferrets differed by a single nucleotide, producing an amino acid alteration. Compared with other types of ADV, these isolates had 96% sequence similarity to a published ferret ADV (FADV) in contrast to <91% homology to various types of mink ADV (MADV). The phylogenetic tree of ADVs indicates that these 2 isolates and the published FADV belong to the same genetic group and definitely are divergent from MADVs. The predicted amino acid sequence of the hypervariable segment in the capsid gene was conserved among the 3 types of FADV. These results indicated that the 2 isolates found in Japan were new DNA types of FADV and could have been derived from FADV(s). A restriction fragment length polymorphism (RFLP) method to distinguish the ferret types of ADV from the mink types of ADV was developed on the basis of differences in their nucleotide sequences. Digestion of the PCR products with Afal or ScaI provided different cleavage patterns for FADV and MADV. This PCR/RFLP analysis of the ADV capsid gene will be a valuable asset for diagnosis of this virus infection in ferrets. PMID- 11478608 TI - Characterization of an undifferentiated malignancy as a mast cell tumor using mutation analysis in the proto-oncogene c-KIT. AB - A 6.5-year-old female Boxer was euthanized and presented for necropsy following rapid clinical decline concomitant with the development of numerous tumor masses. The largest of these masses was in the same location as a mast cell tumor that had been previously removed from this dog. Gross examination revealed the presence of nodules 5-200 mm in diameter throughout the body, including the lymph nodes. Histologic analysis showed an influx of round cells with no granules, leading to the provisional diagnosis of systemic lymphosarcoma. Immunohistochemical staining for B- and T-lymphocyte antigens was negative. Molecular tests were used to identify a tandem duplication in the c-KIT proto oncogene from both the earlier mast cell tumor and the current nodules, implicating a common origin. Addition of molecular testing to conventional necropsy evaluations allowed a definitive diagnosis of mast cell tumors. PMID- 11478609 TI - Digital papillomatosis in a confined Beagle. AB - Papillomavirus-induced papillomas were diagnosed on multiple digits of all 4 feet of a young Beagle. No other cutaneous or oral involvement was identified. Papillomavirus antigen was confirmed by immunoperoxidase localization within keratinocyte nuclei. In addition to the typical basophilic intranuclear inclusions associated with papillomavirus infections, keratinocytes within the papillomas contained large, eosinophilic cytoplasmic inclusions that previously have been described in a Boxer with cutaneous lesions associated with a papillomavirus infection. The papillomas in this Beagle regressed completely within 2 months of the initial diagnosis. PMID- 11478610 TI - Idiopathic disseminated intracytoplasmic neuronal vacuolation in a neonatal Holstein calf born in the USA. AB - Histopathologic, immunohistochemical, and ultrastructural evaluations were made of a 6-day-old Holstein calf with severe vacuolation of the neuronal perikarya that was widely distributed throughout the central nervous system. No evidence of storage material within the vacuoles was revealed by histopathologic and ultrastructural examinations. Immunohistochemical and electron microscopic examinations were negative for protease-resistant prion protein and scrapie associated fibrils, respectively. These results indicate that the clinical signs in this calf were not associated with transmissible spongiform encephalopathy. Neuronal vacuolation has not previously been documented in calves. PMID- 11478611 TI - Rhinosporidiosis in a cat. AB - A polypoid nasal mass from an adult cat was submitted for routine biopsy examination and was found to contain sporangia and sporangiospores consistent with Rhinosporidium seeberi. Inflammatory infiltrates were moderate and pyogranulomatous to lymphohistiocytic and were associated with hyperplasia of the transitional nasal epithelium. Apparently, this is the first reported case of rhinosporidiosis in a cat. PMID- 11478612 TI - Prevalence of eaeA+ Escherichia coli isolated from pigs with diarrhea. AB - A total of 600 Escherichia coli strains isolated from 382 preweaned and 197 postweaned pigs with diarrhea were tested for the presence of the eaeA gene by polymerase chain reaction techniques. Of the 393 isolates from preweaned pigs, 23 (5.8%) E. coli strains isolated from 23 pigs carried the eaeA gene. Of the 207 isolates from postweaned pigs, 9 (4.3%) E. coli strains isolated from 9 pigs carried the eaeA gene. The results suggest that eaeA+ E. coli is associated with diarrhea in pre- and postweaned pigs. PMID- 11478613 TI - Application of competitive enzyme-linked immunosorbent assay for the serologic diagnosis of classical swine fever virus infection. AB - A competitive enzyme-linked immunosorbent assay (C-ELISA), based on a truncated E2 recombinant protein of the Alfort/187 strain of classical swine fever virus (CSFV) and a specific monoclonal antibody M1669, was evaluated using 2,000 sera from clinically healthy pigs in Canada (a CSFV-free country) and sera from experimentally infected pigs. The relative specificity and sensitivity of the C ELISA were 100% and 86%, respectively, at a cutoff of 25% inhibition using negative and positive pig sera, as defined by the neutralizing peroxidase-linked assay (NPLA). A kappa value of 0.91 was obtained, indicating an excellent level of agreement between the NPLA and the C-ELISA. When sera from 120 infected pigs were used in the test at > or = 21 days postinfection, the sensitivity of the C ELISA and the kappa value increased to 97% and 0.98, respectively. This C-ELISA will be useful when a large number of samples must be tested, as could occur during a disease outbreak or for surveillance or prevalence studies. PMID- 11478614 TI - A simpler, more sensitive competitive inhibition enzyme-linked immunosorbent assay for detection of antibody to malignant catarrhal fever viruses. AB - An earlier competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) was developed for detection of specific antibody against malignant catarrhal fever (MCF) viruses (MCFV) in ruminants. In this study, the indirect CI-ELISA was improved by conjugating the monoclonal antibody 15-A directly with horseradish peroxidase and by developing a method of producing precoated, dried antigen plates. This new test is referred to as a direct CI-ELISA. The reformatted test yielded a significantly improved sensitivity, and the time required was reduced to about one-sixth of the previous time. Of 37 MCF cases in cattle that were confirmed by histopathology and polymerase chain reaction (PCR) assay, 37 (100%) were positive by the new test, whereas the indirect CI-ELISA detected only 23 (62%). The direct CI-ELISA detected antibody to MCFV in 100% of 48 sheep that had been defined as infected with ovine herpesvirus 2 (OvHV-2) by PCR, whereas the indirect CI-ELISA detected only 41 (85%). Comparison of antibody titers measured by the 2 assays for sera collected from OvHV-2-infected sheep and from cattle, bison, and deer with clinical sheep-associated MCF revealed that the direct CI ELISA offered a 4-fold increase in analytical sensitivity over the indirect format. The number of seropositive animals detected by the direct CI-ELISA among apparently normal cattle and bison was 2-3 times greater than the number detected by the indirect CI-ELISA, indicating that a significant percentage of normal cattle and bison are subclincally infected with MCFV. PMID- 11478615 TI - Detection and confirmation of reptilian adenovirus infection by in situ hybridization. AB - Adenovirus infections are documented in at least 12 different species of reptiles. In contrast to their mammalian and avian counterparts reptilian adenoviruses are not well characterized as to their pathogenic potential and their ability to cause primary disease. In the diagnostic setting, fresh tissues are often not available for virus isolation, and the confirmation of reptilian adenovirus infections is dependent largely upon electron microscopy for the identification of intranuclear viral inclusions associated with histopathologic changes. The diagnosis of adenovirus infection in 2 different species of snake was confirmed by the application of DNA in situ hybridization. Using an aviadenovirus specific oligoprobe, adenoviral DNA was observed in the nuclei of hepatocytes, Kupffer cells, endothelial cells, and enterocytes. Electron microscopy of the liver confirmed the presence of intranuclear viral particles morphologically consistent with an adenovirus. DNA in situ hybridization on formalin-fixed tissues can serve as a suitable alternative to electron microscopy in the diagnosis of reptilian adenovirus infections. Both affected snakes had other concurrent diseases, suggesting that the adenovirus may not have been the primary pathogen. PMID- 11478616 TI - Treatment of methylacetate waste gas using a trickle-bed air biofilter. AB - The trickle-bed air biofilter (TBAB) performance for methylacetate (MA) removal from waste gases was evaluated under different gas flow rates and influent concentrations. In the pseudo-steady-state conditions, the elimination capacity increased but the removal efficiency decreased with the increase of MA loading. More than 95 and 90% removal efficiencies were achieved for influent MA loadings below 390 and 727 g/m3/h, respectively. The TBAB appears to be a very effective treatment process for controlling MA emission under low to high loading conditions, and the effectiveness could be maintained over 190 days of laboratory operation. PMID- 11478617 TI - Effects of hydrodynamic chromatography on colloid-facilitated migration of radionuclides in the fractured rock. AB - The performance assessment of high level radioactive waste disposal has emphasized the role of colloids in the migration of radionuclides in the geosphere. Previous literature [Nagasaki S, Tanaka S, Suzuki A. Fast transport of colloidal particles through quartz-packed columns. J. Nucl. Sci. Technol. 1975;30(11):1136] indicates that owing to hydrodynamic chromatography the colloid velocity may not be equal to that of groundwater. Using hydrodynamic chromatography, this work investigates the effects of the size of colloidal particles on the radionuclide migration facilitated by colloids in a single fractured porous rock. Also, a methodology is proposed to develop a predictive model to assess transport within the fracture rock as well as various other phenomenological coefficients, particularly the size of colloidal particles. In addition, a fully developed concentration profile for non-reactive colloids in the fracture is developed to elucidate hydrodynamic chromatography of colloids in geological media. The external forces acting on colloidal particles hypothesized in the model proposed herein include inertial force, van der Waals attractive force, double layer force as well as gravitational force. The dispersion coefficient of colloids and the distribution coefficient for radionuclides with colloids are also considered as they pertain to the size of the colloid. In addition, the size distributions of colloids are utilized to investigate the effects of polydispersed colloids. PMID- 11478618 TI - Colloids in the mortar backfill of a cementitious repository for radioactive waste. AB - Colloids are present in groundwater aquifers and water-permeable engineered barrier systems and may facilitate the migration of radionuclides. A highly permeable mortar is foreseen to be used as backfill for the engineered barrier of the Swiss repository for low- and intermediate-level waste. The backfill is considered to be a chemical environment with some potential for colloid generation and, due to its high porosity, for colloid mobility. Colloid concentration measurements were carried out using an in-situ liquid particle counting system. The in-house developed counting system with three commercially available sensors allowed the detection of single particles and colloids at low concentrations in the size range 50-5000 nm. The counting system was tested using suspensions prepared from certified size standards. The concentrations of colloids with size range 50-1000 nm were measured in cement pore water, which was collected from a column filled with a highly permeable backfill mortar. The chemical composition of the pore water corresponded to a Ca(OH)2-controlled cement system. Colloid concentrations in the backfill pore water were found to be typically lower than approximately 0.1 ppm. The specific (geometric) surface areas of the colloid populations were in the range 240 m2 g(-1) to 770 m2 g(-1). The low colloid inventories observed in this study can be explained by the high ionic strength and Ca concentrations of the cement pore water. These conditions are favourable for colloid-colloid and colloid-backfill interactions and unfavourable for colloid-enhanced nuclide transport. PMID- 11478619 TI - A study of iron mineral transformation to reduce red mud tailings. AB - This study examines the effects of iron mineral transformation in an aluminum extraction process on the settling behavior, and the physical and chemical properties of the resulting red mud slurry that must be disposed of. By producing a red mud with a higher solid content, the total volume of mud slurry will also be reduced for a given alumina production rate and more caustic soda will be recovered. The settling behavior and the mineralogical, physical, and physico chemical properties of one bauxite and three red muds processed under varying conditions were analyzed based on examination of the iron mineral transformations. The properties of red muds derived from the same bauxite can differ markedly due to variations in operating conditions of the Bayer process, such as temperature and the addition of a reducing agent. The settling of red mud can be improved by converting goethite into hematite and/or magnetite to produce a mud of larger particle size, smaller specific surface area, and larger specific gravity, characteristics which reduce the total volume of mud slurry to be disposed of and which allow for less potential contamination from caustic soda. This study also found that the by-product--Bayer sodalite--has the high exchange capacity for Na+ that might contribute to the long-term environmental problems. PMID- 11478620 TI - Effect of aluminum sulfate and cationic polyelectrolytes on the destabilization of emulsified wastes. AB - Emulsified oil in wastewater constitutes a severe problem in the different treatment stages. Aluminum salts have been traditionally used as coagulants in wastewater treatments. Polyelectrolytes are used to coagulate and flocculate colloidal systems. The performance of aluminum sulfate in comparison to polyelectrolytes (chitosan and polyacrylamide) as conditioning chemicals for an emulsion waste was tested, and the predominant mechanisms acting in each case were analyzed. Turbidimetry, jar test, colloidal titration and microscopy were used to test emulsion destabilization. Both charge neutralization and bridge formation were identified and confirmed as mechanisms of interaction of polyelectrolytes with waste constitutents. Charge neutralization would be more important for chitosan than for polyacrylamide treatment. A coincidence between the doses necessary to reach zero colloidal charge and minimum turbidity was observed for polyelectrolytes. The time necessary to produce system clarification was larger for aluminum sulfate than for polyelectrolytes; this time was shortened for higher aluminum sulfate concentration. The pH showed a marked effect on aluminum sulfate performance with the optimum at pH 6; polyelectrolyte action was practically not affected by pH. Polyelectrolyte addition produced the minimum turbidity for the same doses that zero colloidal charge; at higher doses, emulsion was restabilized and became turbid again. However, aluminum sulfate treatment did not produce emulsion restabilization. PMID- 11478621 TI - Cement solidification of simulated off-gas condensates from vitrification of low level nuclear waste solutions. AB - Solidification in a cementitious matrix is a viable alternative for low-level nuclear waste management; it is therefore important to understand the behavior and properties of such wasteforms. We have examined the cementitious solidification of simulated off-gas waste streams resulting from the vitrification of low-level nuclear waste. Different possible methods for scrubbing the off-gasses from a vitrifier give rise to three possible types of waste compositions: acidic (from aqueous dissolution of volatile NOx and POx carried over from the vitrifier), basic (from neutralizing the former with sodium hydroxide), and fully carbonated (arising from a direct-combustion vitrifier). Six binder compositions were tested in which ordinary Portland cement was replaced at different proportions by fly ash and/or ground granulated blast furnace slag. A high solution to binder ratio of 1l/1 kg was used to minimize the volume of the wasteform and 10% attapulgite clay was added to all mixes to ensure that the fresh mix did not segregate prior to setting. The 28-day compressive strengths decreased when a high proportion of cement was replaced with fly ash, but were increased significantly when the cement was replaced with slag. The heats of hydration at early age for the various solids compositions decreased when cement was replaced with either fly ash or slag; however, for the fly ash mix the low heat was also associated with a significant decrease in compressive strength. High curing temperature (60 degrees C) or the use of extra-fine slag did not significantly affect the compressive strength. Recommendations for choice of binder formulations and treatment of off-gas condensates are discussed. PMID- 11478622 TI - High temperature co-treatment of bottom ash and stabilized fly ashes from waste incineration. AB - Bottom ashes from two Danish municipal solid waste incineration plants were heated at 900 degrees C with iron oxide stabilized air pollution control residues at actual mass flow ratios (9:1), simulating a treating method for the residues. The two residues were co-treated, producing one combined stream that may be utilized as a secondary road construction material. Scanning electron microscope analysis and grain size distribution analysis indicated that sintering of the particles did not occur. Batch leaching tests at liquid/solid 10 l/kg at a range of pH-values (6-10) quantified with respect to Cd, Cr and Pb revealed significant positive effects of co-heating the ashes, although Pb showed slightly increased leaching. At a liquid/solid ratio of 10 l/kg the leachate concentrations were still low at pH 7-10 and the release of Pb was, thus, not expected to limit the utilization of the mixed ashes. The process, thus, fixates the metals in the solid residues without altering the leaching properties of the bottom ash too significantly. PMID- 11478623 TI - Treatment of anti-osmotic drug based pharmaceutical effluent in an upflow anaerobic fluidized bed system. AB - During the production of Cephradine (a main constituent of anti-osmotic drug) a large quantity of concentrated effluent was produced. The main polluting compounds in this effluent are osmotic drug, acetic acid and ammonia. The feasibility of using a fluidized bed reactor under anaerobic condition with bioaugmentation to treat anti-osmotic drug based pharmaceutical effluent was evaluated. The main objective of the study was to show that bioaugmentation could be used to promote biological treatment to applications where conventional operation might be difficult or unfavourable. The effluent with chemical oxygen demand (COD) of 14000-18000 mg/l was treated in a fluidized bed reactor with a hydraulic retention time of 3-12 h. The reactor was unable to maintain consistent removal in conventional mode of operation due to an inability to retain and grow biomass. The COD reduction (%) after inoculation from a sequencing batch reactor was related to influent concentration, mass of inoculum and hydraulic retention time characterized by calculating the initial food to microorganism ratio. The role of volatile fatty acid (VFA) as cosubstrate was assessed with respect to COD reduction (%). Continuous COD reduction (%) attained a maximum value of 88.5% using bioaugmentation through periodic addition of acclimated cells every 2 days with 30-73.2 g of cells from an off-line enricher reactor. PMID- 11478624 TI - Migration of radionuclides in porous rock in the presence of colloids: effects of kinetic interactions. AB - This work investigates the colloid-facilitated migration of radionuclides with radioactive decay in porous media. The sorption processes for radionuclides with both the solid matrix and colloids are treated as equilibrium or nonequilibrium. An analytical solution is obtained from a simplified linear equilibrium interaction mechanism. In addition, the adsorption processes for radionuclides with colloids and porous rock can be assumed as nonequilibrium and modeled by the linear kinetic adsorption. The numerical method is employed to solve the coupled colloid and radionuclide transport equations under nonequilibrium sorption assumption. Moreover, the reaction rates of the adsorption processes for radionuclides with the solid matrix and colloids affect the transport characteristics of radionuclides. The fast reaction rate of radionuclides with colloids causes a higher concentration of radionuclides adsorbed on colloids in a dispersed phase and enlarges acceleration caused by colloids. However, the fast reaction rate for radionuclides with solid matrix increases the retardation effect caused by the solid matrix. This work developed a predictive model for the transport of colloid-facilitated radionuclides in porous medium and to assess the importance of various phenomenological coefficients, particularly parameters for the adsorption interactions. PMID- 11478625 TI - Investigation of irradiated soil byproducts. AB - The high dose irradiation of windblown soil deposited onto the surface of spent nuclear fuel is of concern to long-term fuel storage stability. Such soils could be exposed to radiation fields as great as 1.08 x 10(-3) C/kg-s (15,000 R/hr) during the 40-year anticipated period of interim dry storage prior to placement at the proposed national repository. The total absorbed dose in these cases could be as high as 5 x 10(7) Gy (5 x 10(9) rads). This investigation evaluated the potential generation of explosive or combustible irradiation byproducts during this irradiation. It focuses on the production of radiolytic byproducts generated within the pore water of surrogate clays that are consistent with those found on the Idaho National Engineering and Environmental Laboratory. Synthesized surrogates of localized soils containing combinations of clay, water, and aluminum samples, enclosed within a stainless steel vessel were irradiated and the quantities of the byproducts generated measured. Two types of clays, varying primarily in the presence of iron oxide, were investigated. Two treatment levels of irradiation and a control were investigated. An 18-Mev linear accelerator was used to irradiate samples. The first irradiation level provided an absorbed dose of 3.9 x 10(5)+/-1.4 x 10(5)Gy (3.9 x 10(7)+/-1.4 x 10(7) rads) in a 3-h period. At the second irradiation level, 4.8 x 10(5)+/-2.0 x 10(5)Gy (4.8 x 10(7)+/-2.0 x 10(7) rads) were delivered in a 6-h period. When averaged over all treatment parameters, irradiated clay samples with and without iron (III) oxide (moisture content = 40%) had a production rate of hydrogen gas that was a strong function of radiation-dose. A g-value of 5.61 x 10(-9)+/-1.56 x 10(-9) mol/J (0.054+/ 0.015 molecules/100-eV) per mass of pore water was observed in the clay samples without iron (III) oxide for hydrogen gas production. A g-value of 1.07 x 10( 8)+/-2.91 x 10(-9) mol/J (0.103+0.028 molecules/100-eV) per mass of pore water was observed in the iron (III) oxide containing clay samples for hydrogen gas production. This value was noticeably larger when the samples were spiked with both KCl and KNO3 salts. The ratio of oxygen to nitrogen gas was observed to increase as a function of absorbed dose particularly in the presence of both KCl and KNO3 salts. The creation of radiolytic byproducts produced an observable but small increase in headspace pressure. Temperature increases during irradiation were not observed. Additionally, KCl and KNO3 salts added to the clays enhanced nitrite production as a function of radiation-dose and the type of clay considered. The addition of aluminum to these samples had no statistically discernable impact at the alpha = 0.05 level. Generation of the irradiation products, hydrogen peroxide and hydrogen gas also depended upon the type of clay irradiated and the presence of both KCl and KNO3 salts and the total dose received. PMID- 11478626 TI - A capillary zone electrophoretic method for the study of formation of a covalent conjugate between microcystin LR and protein phosphatase 2A. AB - The study of conjugate formation between microcystin (MCYST)-LR and protein phosphatase (PP) 2A, which was isolated from bovine kidney and mouse brain, was achieved by using a highly efficient capillary zone electrophoretic (CZE) separation method. The MCYST-LR-PP2A conjugate (from bovine kidney) was resolved from its precursor after just 15 min of incubation. Moreover, the migration time, and, hence, the total analysis time, was less than 5 min. While the present findings of the time lag between conjugate formation and full inhibition are not novel, the CZE method does provide an alternative tool to HPLC with a higher separation efficiency to yield data for kinetic and mechanistic studies of the enzyme-toxin interaction. The CZE data reported here were found not to be adequately described by a first-order kinetic model. Moreover, the CZE method, which involves the use of a low ionic strength aqueous buffer, does not suffer from the drawback of the use of denaturing organic solvents such as those used in HPLC. PMID- 11478627 TI - Simultaneous optimization of the solid-phase extraction of organochlorine and organophosphorus pesticides using the desirability function. AB - In a multi-response situation [solid-phase extraction (SPE) of 18 organochlorine and nine organophosphorus pesticides from aqueous solution], several response variables are considered, and the optimization problem is more complex than in the single response case. Experimental design combined with desirability function was applied for the optimization of the SPE of these pesticides in order to obtain optimum operating conditions for routine work. This multi-criterion approach was developed for the simultaneous optimization of multiple responses and it is simple, easy to apply and permits the user to make subjective judgments on the importance of each response. The desirability function is based on the search for a global optimum [D = f (Y1, Y2, ..., Yn)] by the transformation of the measured property to a dimensionless scale for each criterion. The experimental design strategy involved a central composite design. A compromise between conflicting goals, such as achievement of good yields with SPE recoveries of ca. 100% for a large number of analytes with inherent chemical differences, was found by means of the desirability function D. PMID- 11478628 TI - Arsenic extraction and speciation in carrots using accelerated solvent extraction, liquid chromatography and plasma mass spectrometry. AB - Arsenic present in freeze-dried carrots was extracted using accelerated solvent extraction (ASE). Several parameters, including selection of the dispersing agent, extraction time, number of extraction cycles, particle size and extraction temperature, were evaluated to optimize the ASE method. Filtering and treatment with C-18 SPE cartridges were also evaluated as part of the sample preparation procedure before speciation analysis. The method was validated by spiking single arsenical and mixed arsenical standards on the dispersing agent and on portions of freeze-dried carrot prior to extraction. LC-ICP-MS was used to determine individual arsenic species in the carrot extracts. A weak anion-exchange column was used for the separation of As(III), As(v), monomethylarsonic acid (MMA), dimethylarsinic acid and arsenobetaine. Optimized sample preparation conditions were applied to the extraction of arsenic in nine freeze-dried carrot samples. Total arsenic concentration in the carrot samples ranged from less than 20 ng g( 1) to 18.7 microg g(-1), dry mass. Extraction efficiency, defined as the ratio of the sum of individual arsenic species concentrations to total arsenic, ranged from 80 to 102% for freeze-dried carrots with arsenic concentrations greater than the limit of quantitation. Inorganic As(III) and As(v) were the only species found in samples that contained less than 400 ng g(-1) total arsenic. MMA and an unidentified arsenic compound were present in some of the samples with higher total arsenic content. PMID- 11478629 TI - Regiospecific characterisation of the triacylglycerols in animal fats using high performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry. AB - High performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry (HPLC-APCI MS) was applied to the characterisation of triacylglycerols (TAGs) in animal fats. The major TAGs in four fats (beef, chicken, lamb and pork) were identified and positional isomers assigned according to their APCI mass spectra. Beef and lamb fat TAGs were confirmed as containing higher proportions of saturated fatty acids compared with those of chicken and pork. HPLC-APCI MS was also shown to be of value in providing regiospecific information for the fatty acids in individual TAG species. For example, beef and lamb fat were shown to contain both cis- and trans-isomers of the 18:1 fatty acid, whilst chicken and pork contained only the cis-isomer. When the position of fatty acid substitution was determined from the APCI spectra, whilst the cis- 18:1 was predominantly found in the 2-position of the TAG, the trans-18:1 showed a preference for the 1/3-position. Similarly, it was confirmed that although the 2-position of beef, chicken and lamb fat TAGs was dominated by unsaturated fatty acids, in pork fat, a characteristically high proportion of palmitic acid was seen in this position. The TAGs identified compared well with those reported previously. The distributions of 2-position fatty acids seen in lamb and pork fat compared favourably with those obtained by the more traditional method of lipase degradation. Although the distributions for chicken and beef showed some discrepancies, these can be attributed to weaknesses in the quantification procedure or the specificity of the lipase. Overall, the technique of HPLC-APCI MS has been shown to be very powerful for the regiospecific analysis of animal fats. PMID- 11478630 TI - Exposure to 2,4- and 2,6-toluene diisocyanate (TDI) during production of flexible foam: determination of airborne TDI and urinary 2,4- and 2,6-toluenediamine (TDA). AB - Occupational exposure to 2,4- and 2,6-toluene diisocyanate (2,4- and 2,6-TDI) was measured during the production of flexible foam. The usefulness of urinalysis of the TDI-derived amines, 2,4- and 2,6-toluenediamine (2,4- and 2,6-TDA), for exposure assessment was compared with air monitoring. Urine samples were collected from 17 employees at two plants. The workers' personal exposure was measured using 1-(2-methoxyphenyl)-piperazine (2MP)-impregnated glass fibre filters for sampling and high-performance liquid chromatography (HPLC) with ultraviolet (UV) and electrochemical (EC) detection for quantification. The limit of detection (LOD) of 2,4- and 2,6-TDI was 0.01 microtg ml(-1) for a 20 microl injection. The precision of sample preparation, expressed as the relative standard deviation (RSD), was 0.6% with UV detection and 0.8% with EC detection at a 2,4-TDI concentration of 0.2 microg ml(-1) (n = 6). For 2,6-TDI, the corresponding RSDs were 0.5% and 0.8%. The urinary 2,4- and 2,6-TDA metabolites were determined after acid hydrolysis as heptafluorobutyric anhydride derivatives by gas chromatography-mass spectrometry. The LOD in urine was 0.35 nmol l(-1) for 2,4-TDA and 0.04 nmol l(-1) for 2,6-TDA. The precision (RSD) of six analyses of human urine spiked to a concentration of 100 nmol l(-1) was 3.7% for 2,4-TDA and 3.6% for 2,6-TDA. There was a trend for linear correlation between urinary TDA concentration and the product of airborne TDI concentration and sampling time. Urinalysis of TDA is proposed as a practical method for assessing personal exposures in workers exposed intermittently to TDI. PMID- 11478631 TI - Quick regiospecific analysis of fatty acids in triacylglycerols with GC using 1,3 specific lipase in butanol. AB - A new micro-procedure for the quick regiospecific analysis of triacylglycerols (TAG) with a 1,3-specific lipase, Lipozyme IM 20 (produced from strains of Muccor miehei) is described. After dissolution of triacylglycerols in butan-1-ol or butan-2-ol 250 mg of Lipozyme IM 20 were added and the whole mixture was agitated. This operation was repeated 5-10 times and the esterified fatty acids in the form of butyl esters (FABE) were dissolved in pentane and washed with water. The pentane layer was dried over sodium sulfate and the solvent was removed under vacuum to a constant weight. The residue was analysed using GC-MS. The same triacylglycerols were converted to fatty acid methyl esters (FAME). The FAME were analysed by gas chromatography (GC). PMID- 11478632 TI - Negative ion chemical ionization GC/MS-MS analysis of dialkylphosphate metabolites of organophosphate pesticides in urine of non-occupationally exposed subjects. AB - Low level exposure to organophosphate (OP) pesticides can be determined by the measurement of dialkylphosphate (DAP) metabolites in urine. An analytical method is presented here which can measure the metabolites dimethyl phosphate (DMP), diethyl phosphate (DEP), dimethyl thiophosphate (DMTP), dimethyl dithiophosphate (DMDTP), diethyl thiophosphate (DETP), and diethyl dithiophosphate (DEDTP) at low levels. This was achieved by lyophilization of the urine, derivatization with pentafluorobenzyl bromide (PFBBr) and quantification by negative ion chemical ionization GC/MS-MS. The detection limits for the metabolites were 0.5 microg L( 1) DMP, 0.1 microg L(-1) DEP, 0.1 microg L(-1) DMTP, 0.04 microg L(-1) DMDTP, 0.04 microg L(-1) DETP and 0.02 microg L(-1) DEDTP. The RSD for the analytical method was 4-14% for the six metabolites. The method was used to monitor a group of non-occupationally exposed individuals in Sydney, Australia. The metabolites DMP, DEP, DMTP, DMDTP, DETP and DEDTP occurred in 73, 77, 96, 48, 100 and 2% of the samples with median values of 13, 3, 12, <1, 1 and 1 microg L(-1) respectively. The method is simple to use, sensitive and suitable for routine analysis of non-occupational exposure levels. These detection limits are between one and two orders of magnitude lower than those previously reported in the literature. PMID- 11478633 TI - The signal-to-noise ratio as the measure for the quantification of lysophospholipids by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. AB - The subject of this report is the determination of lysophospholipids; lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylserine and lysophosphatidic acid, by matrix-assisted laser desorption/ionisation time-of flight mass spectrometry (MALDI-TOF MS). The mean signal-to-noise ratio (S/N) was used for the first time as a measure of lysophospholipid concentration. Two different sample preparation procedures were applied, the 'standard' procedure and the 'premix' in order to check to what extent these methods influence the results of the lysophospholipid quantification. Results can be summarised as follows: (a) All classes of lysophospholipids can be easily and sensitively analysed by MALDI-TOF MS. The smallest detectable amount of lysophospholipids was 0.09 pmol on the sample plate. That is about two orders of magnitude lower than the amount detectable by standard chromatographic methods. (b) The mean S/N of all peaks detected in the positive ion mass spectra can be used as a measure of the lysophospholipid concentration. Whereas the S/N for neutral lysophospholipids correlated with the applied concentrations only when the samples were analysed as 'premix', the sample preparation and application procedure did not influence the quantification of acidic lysophospholipids. The standard deviations were not higher than 10% of the mean value. (c) All spectra were additionally analysed in the presence of CsCl. The addition of caesium ions makes the peak identification unambiguous in phospholipid mixtures, but the Cs adducts of lysophospholipids do not properly reflect their concentration and, therefore, they were not useful for quantification. (d) The applicability of the method was demonstrated on the organic extract of human neutrophils. PMID- 11478634 TI - Determination of the abundance of delta15N in nitrate ion in contaminated groundwater samples using an elemental analyzer coupled to a mass spectrometer. AB - A rapid method for measuring the delta15N of nitrate ion in water samples using an isotope ratio mass spectrometer coupled to an elemental analyzer system (EA MS) was investigated. The water should be removed from the analytical sample before measurement with this system. We investigated the application of a super absorbent polymer resin powder to various water samples. Each 1 mg of polymer resin powder can absorb about 50-100 mg of solution depending on the concentrations of major ions. Only samples which contain more than 100 mg l(-1) of nitrate-nitrogen are suitable to be absorbed by the polymer resin for the determination of delta15N of nitrate. Preconcentration by rotary evaporation was necessary for dilute samples but the temperature should be kept below 60 degrees C. The polymer resin (about 8 mg) containing the nitrate was directly analyzed using an EA-MS after being oven-dried at 80 degrees C. Good accuracy (precision +/- 0.3%) for delta15N measurements of nitrate-nitrogen in a sample without any isotope fractionation effects during pre-treatment was observed. Results for delta15N of nitrate in contaminated groundwater samples collected in the spring at a tea plantation area in Shizuoka, Japan, were from 9.8 to 10.6%, which were close to the delta15N abundance in organic fertilizers. PMID- 11478635 TI - Investigation of arsenic speciation in oyster test reference material by multidimensional HPLC-ICP-MS and electrospray tandem mass spectrometry (ES-MS MS). AB - Multidimensional (size-exclusion-anion-exchange-cation-exchange) liquid chromatography with ICP-MS detection was developed to produce a map of water soluble species in an oyster test reference material. The presence of arsenobetaine, trimethyl(2-carboxyethyl)arsonium inner salt, arsenocholine, dimethylarsonic acid, tetramethylammonium ion, As(v) and two arsenosugars was demonstrated by ES-MS-MS. A previously unreported compound was isolated and identified by ES-MS-MS as 5-dimethylarsinoyl-beta-ribofuranose. Anion-exchange chromatography was optimized to produce a chromatographically pure peak of arsenobetaine (accounting for ca. 64% of all water-soluble As present) that was used to quantify this compound. PMID- 11478636 TI - Enantioselective molecular sensing of aromatic amines using tetra-(S)-di-2 naphthylprolinol calix[4]arene. AB - This paper presents new analytical data, and evidence of the fluorescence quenching mechanism involved in enantioselective signals obtained with the chiral host tetra-(S)-di-2-naphthylprolinol calix[4]arene. Excellent selectivity is obtained with phenylglycinol and norephidrine in methanol, and samples of unknown enantiomeric composition have been determined with an average error of 1.08 and 0.62%, respectively (n = 6), on the basis of a single fluorescence measurement. The absence of any quenching observed with the related amines of phenylalaninol is attributed to a methylene spacer which inhibits efficient interaction between the aryl group of the guest and the naphthyl group of the host. The importance of the phenyl group of the guest in the quenching mechanism is confirmed by the complete absence of any quenching effect with cyclohexylethylamine. PMID- 11478637 TI - Enzyme modified microband electrodes: cross-talk effects and their elimination. AB - One microband of an array of four microband electrodes (1 mm long and 25 microm wide with a 25 microm gap) was modified with glucose oxidase by direct electrochemically assisted immobilisation, giving a stable microbiosensor with an apparent Michaelis-Menten constant of 12 mM and an i(max) of 80 nA. Cross-talk effects on the adjacent microbands were studied and three different methods for their elimination were tested: the most efficient one involved catalase deposition on the adjacent microband. Under these conditions, the maximum response at the unmodified microbands was in the worst case about 3% compared with the response of the modified microband. This approach has the potential to fabricate a multianalyte microbiosensor. PMID- 11478638 TI - On-line continuous monitoring of glucose or lactate by ultraslow microdialysis combined with a flow-through nanoliter biosensor based on poly(m phenylenediamine) ultra-thin polymer membrane as enzyme electrode. AB - A miniaturised flow-through biosensor with a cell volume of only a few nanoliters was developed in our laboratory. The biosensor can be directly coupled to a microdialysis or ultrafiltration probe. Sampling and continuous on-line monitoring can thus be carried out at submicroliter levels and as a consequence quantitative recoveries of the analyte of interest are achieved. Via this method excessive calibration procedures, as are necessary with conventional microdialysis, are avoided. Here, the construction and the performance of such a biosensor for the continuous on-line monitoring of glucose and lactate will be presented. The biosensor is based on the amperometric detection of hydrogen peroxide after conversion of the analyte of interest by an immobilised oxidoreductase enzyme. Immobilisation of the enzyme is performed through electropolymerisation of m-phenylenediamine. Strategies to improve the performance (e.g. linearity, selectivity and stability) of the miniaturised biosensor are discussed and ex vivo and in vivo experiments carried out thus far demonstrate the potential of this miniaturised flow-through biosensor. PMID- 11478639 TI - Optical biosensors based on Prussian Blue films. AB - Optical biosensing schemes based on enzymatically modified inorganic/organic transparent films predominately composed of Prussian Blue are demonstrated. The composite film, which is non-electrochemically deposited on a non-conducting support. is used as an optical transducer for flow-through biosensors based on hydrolases and oxidases. Urease and glucose oxidase are utilized as model enzymes. Action of the urea biosensor is based on optical pH sensitivity of Prussian Blue indicator. The glucose biosensor is acting as first-generation optical biosensor based on in situ generated Prussian White transducer for hydrogen peroxide. These simple, single-pass transmission optical biosensors exhibit sensitivity in the millimolar range of concentration. The biosensors are very stable owing to presence of a poly(pyrrolylbenzoic acid) network in the composite material. This organic polymer plays a dual role as a binding agent for inorganic material and as a functionalized support for strong covalent immobilization of enzyme molecules. PMID- 11478640 TI - An unmediated hydrogen peroxide biosensor based on hemoglobin incorporated in a montmorillonite membrane. AB - Hemoglobin was incorporated in a montmorillonite membrane. Electrochemical and spectroscopic studies revealed that the electron transfer reactivity and peroxidase activity of the protein were both enhanced. Nevertheless, its structure was still maintained native-like in the membrane. An unmediated hydrogen peroxide biosensor was accordingly prepared. The calibration plot of this H202 sensor was linear in the range of 1.0 x 10(-6)-6.0 x 10(-4) mol L(-1). The relative standard deviation was 3.1% for six successive determinations at a concentration of 1.0 x 10(-4) mol L(-1). The detection limit was 6.0 x 10(-7) mol L(-1). Possible interferences in real sample analyses are discussed. PMID- 11478641 TI - Construction and application of phenytoin anion-selective electrode based on bulk acoustic wave (BAW) sensing technique. AB - This paper describes the construction of an anion-selective bulk acoustic wave (BAW) sensor for the direct determination of phenytoin sodium for the first time. Based on the sensitive mass response of a piezoelectric quartz crystal (PQC) and selective adsorption-desorption across the modified film, the BAW sensor was fabricated by coating a polyvinylchloride (PVC) film containing activator on one side of a PQC. The method was found to be sensitive, rapid and easy to handle without pretreatment of the sample. The logarithm of the frequency shift of the PQC shows a linear relationship to the logarithm of the concentration of phenytoin over the range 6.7 x 10(-8)-8.0 x 10(-4) M with a detection limit of 1.0 x 10(-8) M at pH 10.0. Recoveries were in the range 98.5-102.0%. Influencing factors were examined and optimized. Also discussed is the preliminary application of the phenytoin BAW sensor in serum and injection. The results for real samples obtained by the proposed method agreed with those obtained by conventional methods. PMID- 11478642 TI - Electrochemiluminescence of tris(2,2'-bipyridine)ruthenium(II) immobilized in poly(p-styrenesulfonate)-silica-Triton X-100 composite thin-films. AB - The electrochemiluminescence (ECL) of tris(2,2'-bipyridine)ruthenium(II) [Ru(bpy)3(2+)] immobilized in poly(p-styrenesulfonate) (PSS)-silica-Triton X-100 composite films was investigated. The cooperative action of PSS, sol-gel and Triton X-100 attached Ru(bpy)3(2+) to the electrode strongly, and the presence of Triton X-100 prevented drying fractures of the sol-gel films during gelation and even on repeated wet-dry cycles. The modified electrode was used for the ECL detection of oxalate, tripropylamine (TPA) and NADH in a flow injection analysis (FIA) system with a newly designed flow cell. The detection scheme exhibited good stability, short response time and high sensitivity. Detection limits were 0.1, 0.1 and 0.5 micromol L(-1) for oxalate. TPA and NADH, respectively, and the linear concentration range extended from 0.001 to 1 mmol L(-1) for the three analytes. Applications of the flow cell in ECL and electrochemical detection, as well as the immobilization of reagents based on the cooperative action, are suggested. PMID- 11478643 TI - Investigation of microcontact transfer of proteins from a selectively plasma treated elastomer stamp by fluorescence microscopy and force microscopy. AB - Selective plasma treatment of the recessed regions of the elastomer stamps is shown to alter the resultant protein patterns. Fluorescence microscopy is demonstrated to be an excellent tool to discriminate between regions of microcontact printed fluorescent dye-labelled albumin in polystyrene. Atomic force microscopy and shear force microscopy are used to provide high-resolution images of the patterned protein layers. The formation and characteristics of the patterns formed by these alternative strategies is discussed. PMID- 11478644 TI - Sustained prediction ability of net analyte preprocessing methods using reduced calibration sets. Theoretical and experimental study involving the spectrophotometric analysis of multicomponent mixtures. AB - A newly developed multivariate method involving net analyte preprocessing (NAP) was tested using central composite calibration designs of progressively decreasing size regarding the multivariate simultaneous spectrophotometric determination of three active components (phenylephrine, diphenhydramine and naphazoline) and one excipient (methylparaben) in nasal solutions. Its performance was evaluated and compared with that of partial least-squares (PLS 1). Minimisation of the calibration predicted error sum of squares (PRESS) as a function of a moving spectral window helped to select appropriate working spectral ranges for both methods. The comparison of NAP and PLS results was carried out using two tests: (1) the elliptical joint confidence region for the slope and intercept of a predicted versus actual concentrations plot for a large validation set of samples and (2) the D-optimality criterion concerning the information content of the calibration data matrix. Extensive simulations and experimental validation showed that, unlike PLS, the NAP method is able to furnish highly satisfactory results when the calibration set is reduced from a full four-component central composite to a fractional central composite, as expected from the modelling requirements of net analyte based methods. PMID- 11478645 TI - Robust linear regression taking into account errors in the predictor and response variables. AB - We developed a robust regression technique that is a generalization of the least median of squares (LMS) technique to the field in which the errors in both the predictor and the response variables are taken into account. This simple generalization is limited in the sense that the resulting straight line is found by using only two points from the initial data set. In this way a simulation step is added by using the Monte Carlo method to generate the best robust regression line. We call this new technique 'bivariate least median of squares' (BLMS), following the notation of the LMS method. We checked the robustness of the new regression technique by calculating its breakdown point, which was 50%. This confirms the robustness of the BLMS regression line. In order to show its applicability to the chemical field we tested it on simulated data sets and real data sets with outliers. The BLMS robust regression line was not affected by many types of outlying points in the data sets. PMID- 11478646 TI - Quantitative determination of binary and tertiary calcium carbonate mixtures using powder X-ray diffraction. AB - The ability to determine the calcium carbonate polymorphic ratio of calcite, aragonite and vaterite in a mixture is important for a variety of applications, particularly the fields of biomineralisation and crystal engineering. Raman spectroscopy and powder X-ray diffraction were used to quantitatively determine the polymorphic composition of both binary and tertiary mixtures of calcium carbonate. It was found that the quantitative detection limits of powder X-ray diffraction were superior to both Raman and infrared spectroscopy. PMID- 11478647 TI - Near infrared reflectance spectroscopy for the fast identification of PVC-based films. AB - Near infrared (NIR) reflectance spectroscopy was used to develop a non destructive and rapid qualitative method for the analysis of plastic films used by the pharmaceutical industry for blistering. Three types of films were investigated: 250 microm PVC [poly(vinyl chloride)] films, 250 microm PVC films coated with 40 g m(-2) of PVDC [poly(vinylidene dichloride)] and 250 microm PVC films coated with 5 g m(-2) of TE (Thermoelast) and 90 g m(-2) of PVDC. Three analyses were carried out using different pre-treatment options and a PLS (partial least squares) algorithm. Each analysis was aimed at identifying one type of film and rejecting all types of false sample (different thickness, colour or layer). True and false samples from four plastics manufacturers were included in the calibration sets in order to obtain robust methods that were suitable regardless of the supplier. Specificity was demonstrated by testing validation sets against the methods. The tests showed 0% of type I (false negative identification) and 1% of type II errors (false positive identification) for the PVC method, 13 and 3%, respectively, for the PVC-PVDC method and no error for the PVC-TE-PVDC method. Type II errors, mostly due to the slight sensitivity of the methods to film thickness, are easily corrected by simple thickness measurements. This study demonstrates that NIR spectroscopy is an excellent tool for the identification of PVC-based films. The three methods can be used by the pharmaceutical industry or plastics manufacturers for the quality control of films used in blister packaging. PMID- 11478648 TI - Influence of the procedure used to prepare the calibration sample set on the performance of near infrared spectroscopy in quantitative pharmaceutical analyses. AB - Calibrating near infrared diffuse reflectance spectroscopy (NIRS) methods usually involves preparing a set of samples with a view to expanding the analyte concentration range spanned by production samples. In this work, the performances of the two procedures most frequently used for this purpose in near infrared pharmaceutical analysis, viz., synthetic samples obtained by weighing of the pure constituents of the pharmaceutical and doped samples made by under- or overdosing previously powdered production samples, were compared. Both procedures were found to provide similar results in the quantification of the active compound in the pharmaceutical, which was determined with a relative standard error of prediction (RSEP) of < 1.6%. However, the two types of sample preparation provide different spectra, which precludes the accurate quantification of synthetic samples from calibrations obtained with doped samples and vice versa. None of the mathematical pre-treatments tested with a view to reducing this different scattering (viz., second derivative, standard normal variate and orthogonal signal correction) could effectively solve this problem. This hinders accurate validation of the linearity of the procedure and makes it advisable to use doped samples which are markedly less different to production samples. PMID- 11478649 TI - Multi-component kinetic-spectrophotometric analysis. Selection of wavelength and time ranges. AB - An empirical method for the selection of the best wavelength and time ranges which can be used in the quantification of binary mixtures, in a kinetic spectrophotometric system, is proposed. It is based on finding those ranges which provide the least correlation between the kinetic profiles and the spectra of the products of reaction. The method was applied to the analysis of binary mixtures using simulated data with different rate constant ratios and in the presence of an interference that shows spectral overlap with the analytes. Subsequently, the proposed method was applied to the resolution of dyphylline and proxyphylline mixtures. The system studied was characterized by an elevated similarity in the kinetic behavior of the analytes under pseudo-first-order conditions and an elevated degree of spectral overlap of the products of reaction. In spite of this, satisfactory results were obtained in the quantification of the two analytes. The standard error of prediction (SEP) and the standard deviation between replicates (SDBR) did not show significant differences, being of the order of 4 and of 3% for dyphylline and proxyphylline, respectively. PMID- 11478650 TI - Determination of the content and identity of lidocaine solutions with UV-visible spectroscopy and multivariate calibration. AB - A method is proposed for the determination of the content and identity of the active compound in pharmaceutical solutions by means of ultraviolet-visible (UV Vis) spectroscopy, orthogonal signal correction (OSC) and multivariate calibration with soft independent modelling of class analogy (SIMCA) classification and partial least squares (PLS) regression. The content was determined with PLS regression and the identity with PLS regression and SIMCA classification. The method was tested on the local anaesthetic compound lidocaine. For the validation, external test sets of both manufactured sample solutions and samples from a stability study were used. For comparison with this new method, liquid chromatography was used as a reference method. The results show that in respect of accuracy, precision and repeatability, the new method is comparable to the reference method. The main advantage over liquid chromatography is the much shorter time of analysis and the simpler analytical procedure. An estimate of the analysis time saved with the proposed method compared with using liquid chromatography, together with practical considerations, is given. PMID- 11478651 TI - Simultaneous determination of Cd(II), Cu(II) and Pb(II) in surface waters by solid phase extraction and flow injection analysis with spectrophotometric detection. AB - A method for heavy metal monitoring using spectrophotometric detection is presented. Traces of Cu(II), Pb(II) and Cd(II) at the low microg l(-1) level can be determined simultaneously after both selective removal of metal interferences and preconcentration using 'extraction chromatographic resins'. Lewatit TP807'84, which contains di(2,4,4-trimethylpentyl)phosphinic acid as active component, was used as solid adsorbent. Two minicolumns containing this resin were used: one at pH 3.2 for the removal of interferences, such as Zn(II) and Fe(III), and the other at pH 5.5 for the selective preconcentration of the target analytes. Spectrophotometric determination used FIA methodology with sulfarsazene as chromogenic reagent and partial least-squares multivariate calibration. The method was successfully applied to the analysis of surface waters from the Llobregat river and ground water samples from wells in the Guadiamar basin. Accuracy, expressed in terms of recoveries, was in the range 80-120% and relative standard deviations were below 10%. PMID- 11478652 TI - Spectrophotometric determination of trace molybdenum in plants and seeds with 3,5 dibromo-4-hydroxyphenylflurone. AB - A novel spectrophotometric method based on a new reagent, 3,5-dibromo-4 hydroxyphenylflurone, was developed for the determination of molybdenum in plants and seeds. 3,5-Dibromo-4-hydroxyphenylflurone showed outstanding analytical characteristics for spectrophotometric determination of molybdenum. The reaction conditions are simple and stable. In 0.2 mol l(-1) phosphoric acid medium (which can combine with iron and other metal ions and greatly improves the selectivity of the color system), molybdenum(VI) reacts with 3,5-dibromo-4 hydroxyphenylflurone to form a 1:2 red complex with an absorption maximum at 530 nm, the color reaction can completed in 2 min and the absorbance of the molybdenum complex remains stable for at least 72 h at room temperature. Its stability constant is 1.21 x 10(28) at 25 degrees C. Beer's law is obeyed over the range 0-0.6 microg ml(-1) Mo(VI). The reagent has very high sensitivity and selectivity; the molar absorptivity of the complex is 1.35 x 10(5) 1 mol(-1) cm( 1) and the limit of quantification, the limit of detection and relative standard deviation (n = 10) were found to be 6.7 ng ml(-1), 2.2 ng ml(-1) and 1.01%, respectively. Cu (50000-fold), Fe (20000-fold), K (20000-fold), NH4+ (20000 fold), Mg (15000-fold), Zn (10000-fold), Na (10000-fold), Al (4000-fold), Ca (25000-fold), Mn (2000-fold), Ce (500-fold), Cr (400-fold) and Bi (200-fold) do not interfere with the determination of trace levels of molybdenum up to the excesses indicated. The selectivity is much superior to that of other published methods. The proposed method was applied to the direct determination of molybdenum in plants and seeds with satisfactory results. The synthesis of the reagent and conditions of color reaction were studied in detail. PMID- 11478653 TI - 1-(2-hydroxy-3-methoxybenzylideneamino)-8-hydroxynaphthalene-3,6-disulfonic acid as a reagent for the spectrophotometric determination of boron in ceramic materials. AB - A sensitive and selective spectrophotometric method for the determination of boron is described. The method is based on the colour reaction between boron and the reagent 1-(2-hydroxy-3-methoxybenzylideneamino)-8-hydroxynaphthalene-3,6 disulfonic acid (HMOA). In a HOAc-NH4OAc buffer of pH 5.5, HMOA reacts with boron to form a 1:2 yellow complex with a maximum absorption at 423 nm. The absorbance (lambdamax = 423 nm) is linear up to 1.2 microg ml(-1) boron in aqueous solution with a repeatability (RSD) of 1.12%. The molar absorptivity and Sandell's sensitivity are 7.19 x 10(3) l mol(-1) cm(-1) and 0.0015 microg cm(-2), respectively. The limit of quantification and limit of detection were found to be 17.1 and 5.2 ng ml(-1), respectively. The interference of various ions was examined in detail. All the metal ions studied can be tolerated in considerable amounts; in particular, the tolerance limits of Fe, Al, Zn, Ca and Mg are superior to those of other reagents such as Azomethine-H and Azomethine-HR. The proposed method was applied to the determination of boron in ceramic materials with satisfactory results. PMID- 11478654 TI - Highly sensitive spectrofluorimetric determination of trace amounts of lead with a new fluorescent reagent, 2-hydroxy-1-naphthaldehydene-8-aminoquinoline. AB - A new fluorescent reagent, 2-hydroxy-1-naphthaldehydene-8-aminoquinoline (HNAAQ), was synthesized. The fluorescent reaction of this reagent with lead was also studied. Based on this chelation, a highly sensitive spectrofluorimetric method was developed for the determination of trace amounts of lead in a water-ethanol (5 + 1, v/v) medium at pH = 9.4. Under these conditions, the Pb-HNAAQ complex has excitation and emission maxima at 398 and 450 nm, respectively. The linear range of the method is from 0 to 100 microg l(-1) and detection limit is 0.28 microg l( 1) of lead. Interference of other ions was studied. In strongly basic media most interfering metal ions form precipitates of hydroxides or oxides and can be removed efficiently through filtration while the residual cations can be removed with a cation-exchange resin. Hence, the selectivity of the method can be increased considerably. The procedure can easily be performed and affords good precision and accuracy. The method was successfully applied to the determination of lead in wheat and rice flour. PMID- 11478655 TI - Study on the co-luminescence system of Dy-Gd-1,6-bis(1'-phenyl-3'-methyl-5' pyrazol-4'-one)hexanedionecetyltrimethylammonium bromide and its analytical application. AB - Dy-1,6-bis(1'-phenyl-3'-methyl-5'-pyrazol-4'-one)hexanedione cetyltrimethylammonium bromide (Dy-BPMPHD-CTMAB) ion association system has strong fluorescence intensity. In this system, some rare earth ions such as Gd3+, Y3+ and La3+ can exert a fluorescence enhancement effect, leading to a newly found co-luminescence system. From this, a rapid, simple and sensitive method was developed for the determination of trace amounts of Dy3+. The results indicate that the fluorescence intensity of the system is linearly related to the concentration of Dy3+ in the range 1.0 x 10(-7)-1.2 x 10(-5) mol L(-1) and the detection limit (S/N = 3) is 3.0 x 10(-8) mol L(-1). The luminescence mechanism of the system is discussed. PMID- 11478656 TI - Spectrophotometric determination of trace aluminium content in parenteral solutions by combined cloud point preconcentration-flow injection analysis. AB - A cloud point preconcentration and flow injection (FI) analysis methodology for aluminium(III) determination has been developed. The analyte in the initial aqueous solution was complexed with Chrome Azurol S (CAS) in the presence of the cationic surfactant benzyldimethyltetradecylammonium chloride (BDTAC). The absorption spectroscopic characteristics of the ternary complex [Al(III)-CAS BDTAC] were examined in detail. The preconcentration step was carried out by means of the non-ionic surfactant polyethylene glycol p-nonylphenyl ether (PONPE 7.5). The enriched analyte solution was injected into an FI system using an HPLC pump. The chemical variables affecting the analytical performance of the combined methodology were studied and optimised. The developed approach was successfully applied to the determination of trace amounts of aluminium in parenteral solutions without previous treatment. Under the optimum experimental conditions, 99.9% extraction was achieved for a preconcentration factor of 50. The limit of detection was 1.12 x 10(-7) mol(-1). The calibration plot was linear over at least two orders of magnitude of aluminium concentration. The developed coupled methodology, which thoroughly satisfies the typical requirements for pharmaceutical control processes, is appropriate for monitoring the aluminium concentration in parenteral nutrition. PMID- 11478657 TI - Two-parameter determination in vinegar by a flow injection-pervaporation system. AB - A flow injection method (FI) for the sequential determination of ethanol and acetic acid in vinegar is reported. The determination of ethanol is based on the oxidation of the pervaporated ethanol by K2Cr2O7. The acetic acid is determined by an acid-base reaction with Thymol Blue as the indicator. Both reactions are monitored photometrically at 600 nm using a single detector. Optimisation studies and assessment of the sequential Fl method are also reported. The linear determination range is 0-12% v/v for ethanol and 0-10% (grams of acetic acid in 100 ml) for acetic acid. The sample throughput of the sequential manifold is seven per hour. The new method was applied to vinegar samples and the results obtained were in excellent agreement with those from reference methods used in Spain. PMID- 11478658 TI - Recent extraction techniques for solid matrices-supercritical fluid extraction, pressurized fluid extraction and microwave-assisted extraction: their potential and pitfalls. PMID- 11478659 TI - Practical approach to archival and retrieval of analytical data in the laboratory. AB - Today's analytical laboratory uses a large number of different instruments that are connected in networks. Together with increasing automation, data are produced at a rate that can easily reach gigabytes per month, which generates the problem of systematic archival. In addition, working under Good Laboratory Practice requires that archival of raw data be performed in such a way that they can be readily retrieved upon request, even years later. While systematic archival of data is already performed in most laboratories, it is the retrieval of saved information that is often far from straightforward. This paper describes a simple but systematic approach for both archival and retrieval of data files and related electronic documents. It consists of an unambiguous scheme for the naming of electronic files, an efficient backup strategy, a simple database holding information about any data acquired, and a convenient interface to this database that can be accessed from any workplace while assuring restricted access. The system is capable of handling several databases concurrently and is used in our facility to archive data from several workgroups. The use of freely available software such as the Linux operating system made it possible to implement a fast and stable solution at exceptionally low cost. PMID- 11478660 TI - Colorimetric determination of inorganic iodine in fortified culinary products. AB - The presented colorimetric procedure only requires simple laboratory equipment and is suitable as a routine procedure for checking concentrations of iodine in fortified culinary products. The Moxon and Dixon colorimetric procedure for iodine determination has been optimised for the determination of iodide and iodate in fortified culinary products, always containing high salt levels. The high sensitivity of the method permits a high dilution of the product solutions, thus reducing interferences from the inherent colour of the products. The calibration is linear in the range from 0 to 12 microg L(-1) of iodine with R2 > 0.99. A series of commercial culinary products were used to validate the method. Recoveries of iodine, added as iodide and/or iodate, were generally in the range 100+/-10%. High concentrations of chloride are essential to obtain a complete recovery of iodate. Limit of quantification was estimated to be 2 mg kg(-1) of product, based on 2-3 g of product. Concentrations of iodine determined with this method were similar to those obtained by an ICP-MS procedure. PMID- 11478661 TI - Successive non-suppressed ion chromatography of common cations and anions using dual columns with single eluent. AB - A successive non-suppressed ion chromatography (IC) system for the determination of common cations (Na+, K+, Mg2+, Ca2+) and anions (Cl-, Br-, NO3-, SO4(2-)) was developed, using two separation columns and a single eluent. 5-Sulfoisophthalic acid eluent was very suitable for such separations with a commercially available cation-exchange column for the mono- and di-valent cations and with an ODS column coated with cetyltrimethylammonium for the anions. Both cations and anions were detected with conductimetrically high sensitivity without any suppressor. After injecting an aliquot of sample solution, the solvent front from the cation exchange column, including most of the anionic species, was firstly accumulated into the additional 2 ml accumulation loop for 60 s, while the cation IC was performed. Subsequently, the accumulated fraction was introduced into the anion exchange column and chromatographed. Relative standard deviations (RSDs) of retention times and conductimetric area responses for common cations were within 6% and within 4%, respectively. The linear relationships between molar concentration and detector response ranged from 0.01 to 1.00 mM with r2 of 0.9994 for Na+, 0.9992 for K+, 0.9993 for Mg2+, and 0.9988 for Ca2+. The successive anion IC through the accumulating process was also quantitative, with 95% recovery or over for each analyte. The linear ranges were between 0.01 and 1.00 mM with r2 of 0.9996 for Cl-, 0.9997 for Br-, 0.9993 for NO3-, and 0.9984 for SO4(2-). The method was applied to the determination of common cations and anions in several mineral waters and a hot spring water. PMID- 11478662 TI - Separation and determination of denatured caseins by hydrophobic interaction chromatography Part II. Method validation and applications. AB - A method recently described for the separation of denatured alpha-, beta- and kappa-caseins by hydrophobic interaction chromatography was validated by the analysis of reference skim milk powder (BCR-063R) certificated for total nitrogen content. The method is based on fast and easy solubilization of commercial and real samples by 4.0 M guanidine thiocyanate and elution on a TSK-Gel Phenyl-5PW column (TosoHaas) in the presence of 8.0 M urea in the mobile phase. No preliminary precipitation or separation of the casein fraction is required. A linear relationship between the concentration of casein and peak area (UV absorbance detector at 280 nm) was obtained over the concentration range 0.5-60 microM. The detection limits for alpha-, beta- and kappa-caseins ranged between 0.30 and 0.65 microM. The precision of the method was evaluated; the relative standard deviation for alpha-, beta- and kappa-casein determination ranged between 2.2 and 2.7% for standard solutions and between 3.5 and 6.2% for real sample solutions. The mean casein content found in 10 aliquots of BCR-063R calculated with respect to the total protein content (estimated on the basis of certified total nitrogen content) was 79.1+/-2.7%. Results of linear fitting of standard additions data for alpha-, beta- and kappa-caseins to BCR-063R were compared with linear fitting of alpha-, beta- and kappa-casein calibration data. The method was applied to commercial caseins and to 31 real, raw samples [processed cow's milk (pasteurised, UHT-treated), follow-up milk powders, cream, cheeses, casein-free infant formulae, cookies for babies containing milk proteins] with the aim of showing the wide applicability of the method in order to determine alpha-, beta- and kappa-caseins. PMID- 11478663 TI - Chromatin-bound cardiolipin: the phospholipid of proliferation. AB - Chromatin-bound lipids, cardiolipin (CL), diglycerides, cholesterol, and cholesterol esters, together with nonhistone proteins, play a key role in structural and functional organization of the chromatin genome during various stages of evolution. There are two pools of chromatin lipids, namely loosely- and tightly-bound lipids. The entire chromatin cardiolipin is bound to DNA. The CL molecule has a common 'interphosphate' structural motive with DNA, i.e. DNA and CL phosphate moieties separated from each other with six chemical bonds and equidistant, which is important for CL functional role, the regulation of gene expression. The CL dominates in the DNA of the active genome but not in the DNA of the repressed genome. The amount of CL in the DNA from the repressed genome of pigeon erythrocytes (one CL molecule per 20 nucleosomes) is 20 times less than in the DNA from the active genome of rat thymus and liver and in the DNA of transformed cells. Cardiolipin provides A-form DNA in the complex with RNA polymerase, which is necessary for transcription. The biological and structural function of cardiolipin can be realised only when unsaturated fatty acyl residues are present in its structure. PMID- 11478664 TI - Translocation (3;12) (p21-pter; q24.1-qter) and phenylketonuria. AB - Cytogenetic karyotyping in mental retardation associated with physical dysmorphism has been regarded as the primary key for the classification of syndromes and other genetic disorders for the predisposition of neoplasia and other fatal diseases. Giemsa-banding of metaphase chromosomes in lymphocytes is a traditional and routine process for the identification of the chromosomal counterpart which can provide a clue for molecular investigation in the subject. An 8-year-old girl showed a diploid karyotype 46, XX, t(3;12) (p21-pter, q24.1 qter) in peripheral blood lymphocyte culture. Biochemical examination of urine labelled her as a case of phenylketonuria. The maternal karyotyping was similar and confirmed the maternal transmission of the translocation. PMID- 11478665 TI - The surface energy of water: the largest but forgotten source of energy in biological systems. AB - Many functional proteins perform mechanical, structural or chemical work. Such proteins often use the energy from the hydrolysis of adenosine triphosphate (ATP). The role of ATP as an energy source and its production by metabolism was established in the middle of the twentieth century and replaced glycolysis as the focus of study. Before this time the surface energy of water, quantified in the middle of the nineteenth century, had been visualized as an important source of biological energy. Experimental and theoretical work has shown that the internal work done by this energy source may greatly exceed the energy derived from metabolism. Although the energy from ATP usually does the work external to the body, even this may be supplemented by the surface energy of water to give greater efficiency. The consideration of the principles by which proteins might employ this larger source of energy to do work is germane at this time. PMID- 11478666 TI - Melioidosis: a clinical model for gram-negative sepsis. PMID- 11478667 TI - A comparison of the isolation rates of Salmonella and thermophilic Campylobacter species after direct inoculation of media with a dilute faecal suspension and undiluted faecal material. AB - Regardless of media used, dilution of faecal samples before direct plating may improve isolation rates and reduce subcultures by freeing organisms from the faecal mass and diminishing competing flora. Despite the routine use of dilution in many laboratories, it has never been established properly whether direct or dilute inocula should be used in primary plating of faeces. A total of 3764 faecal samples was examined in four laboratories with a standardised methodology. The isolation rates, competing flora and confirmatory work performed for Salmonella spp. and Campylobacter spp. from primary plating media with a dilute faecal inoculum were compared with those after direct inoculation of faecal material. Inoculum effects on the isolation of Shigella spp. could not be assessed as only one isolate occurred during the study period. The overall isolation rates of both major enteric pathogens were unaffected by the inoculum. However, significantly fewer wasted subcultures were recorded with a dilute inoculum for Campylobacter spp., and competing florawas reduced in all cases without diluting out small numbers of the pathogen. PMID- 11478668 TI - Evaluation of three methods to measure anti-Brucella IgM antibodies and interference of IgA in the interpretation of mercaptan-based tests. AB - The results of a dipstick assay for the detection of immunoglobulin M (IgM) to Brucella smooth lipopolysaccharide (S-LPS) correlated with those of an enzyme linked immunosorbent assay (ELISA) for IgM and of the serum agglutination test (SAT) performed with and without dithiothreitol. Two sera which were dithiothreitol-sensitive and were dipstick negative were shown to contain specific IgA. The dipstick assay is recommended as a simple method for detecting specific IgM antibodies in acute-phase brucellosis patients. PMID- 11478669 TI - Sensitivity and performance characteristics of a direct PCR with stool samples in comparison to conventional techniques for diagnosis of Shigella and enteroinvasive Escherichia coli infection in children with acute diarrhoea in Calcutta, India. AB - As the sensitivity of the conventional techniques for identifying Shigella spp. and enteroinvasive Escherichia coli (EIEC) causing dysentery cases is low, a PCR assay was evaluated in this study. Analytical sensitivity (2 x 10(2) cfu) of the PCR technique was obtained by artificially spiking negative stool samples with a standard strain of S. flexneri type 2, then determining the detection limit. Specificity (100%) of the method was determined by testing a number of known Shigella and EIEC strains and organisms other than Shigella spp. A total of 300 stool samples collected from children with acute diarrhoea was plated on to two selective agar media after enrichment in Luria broth. Shigella spp. were isolated from 7.7% (23 of 300) and EIEC from 1% (3 of 300) patients. All enriched stool samples were subjected to PCR to amplify the target sequence of invasive plasmid antigen (ipa)H locus, a multicopy element found on the chromosome and invasion plasmid. The stool PCR was positive in 24 of the 26 culture-positive and in 22 culture-negative stools, thus detecting the presence of Shigella spp. or EIEC in 15.3% (46 of 300) of diarrhoea cases. When an ial probe was used for colony hybridistion with enriched stool cultures blotted on to membranes, 9.6% (29 of 300) of dysentery cases were identified as being caused by Shigella spp. or EIEC. Thus the sensitivity of enriched stool culture, colony hybridisation and enriched stool PCR was found to be 54%, 60% and 96%, respectively, when each of the methods was compared to the total microbiologically confirmed cases of dysentery. It was also observed that only 38% (48 of 126) of acute bloody dysentery cases actually had shigella or EIEC infection, as confirmed by laboratory methods. Moreover, this PCR assay could identify a number of untypable Shigella strains (Sh OUT), which would have remained undiagnosed had this assay not been used. PMID- 11478670 TI - Identification of catalase-like activity from Mycobacterium leprae and the relationship between catalase and isonicotinic acid hydrazide (INH). AB - As Mycobacterium leprae proliferate inside macrophages, it has been speculated that catalase encoded by katG may protect the bacilli from deleterious effects of peroxide generated from the macrophage and may also play a crucial role in the survival of M. leprae in vivo. However, unlike that of M. tuberculosis, the katG of M. leprae has been reported to be a pseudogene, implicating that isoniazid, which is activated to a potent tuberculocidal agent by catalase, is unlikely to be of therapeutic benefit to leprosy patients. These results raise a question as to how M. leprae avoids H202-mediated killing inside macrophages. To understand the survival of M. leprae in macrophages, the present study attempted to detect catalase-like activity in M. leprae. Catalase-like activity was found in M. leprae cell lysate by the diaminobenzidine (DAB) staining method with non denaturing polyacrylamide gel electrophoresis. An ammonium sulphate precipitation study revealed that the catalase-like activity was precipitable with 80% ammonium sulphate. The effect of isoniazid (INH) on M. leprae growth was also tested by RT PCR and radiorespirometric assay to examine catalase-like activity in M. leprae, because INH was activated by catalase. It was found that the viability of M. leprae was decreased at a concentration of 20 microg/ml by radiorespirometric assay and it was inhibited at higher concentrations as determined by RT-PCR. These data suggest that a catalase-like activity other than that encoded by katG is present in M. leprae. PMID- 11478671 TI - Characterisation of VanA and VanB elements from glycopeptide-resistant Enterococcus faecium from Greece. AB - Ten glycopeptide-resistant Enterococcus faecium isolates from separate patients in Laikon General Hospital, Athens were studied. Eight isolates had the VanA phenotype and represented variants of three strains based on SmaI macrorestriction banding patterns. Their VanA elements were compared with the prototype element, Tn1546, by an overlapping PCR method. Three related isolates contained resistance elements indistinguishable from Tn1546 (designated Greek type I). The other five isolates all contained identical elements that differed from Tn1546 by the presence of IS1251 between vanS and vanH, by a point mutation (G --> T) at nucleotide position 8234 within vanX and by a partial loss of transposition gene orf1 (designated Greek type II). Two distinct strains of E. faecium with the VanB phenotype were obtained. HhaI digestion of an amplified fragment of the vanB gene indicated that both strains contained the vanB2 allele, and further PCR assays confirmed that the vanB2 gene cluster was located within a Tn5382-like element. PMID- 11478672 TI - Role of T cells in granuloma formation induced by Rhodococcus aurantiacus is independent of their interferon-gamma production. AB - Intravenous injection of Rhodococcus aurantiacus into mice causes granulomatous inflammation dependent on endogenous interferon-gamma (IFN-gamma). This study investigated the mechanism of granuloma formation with an adoptive transfer system in IFN-gamma knockout (IFN-gamma(-/-)) mice. IFN-gamma(-/-) mice infected with R. aurantiacus did not develop granulomas, and high titres of endogenous interleukin-10 (IL-10) were detected in spleen extracts at 2 weeks after infection. The adoptive transfer of splenocytes from infected wild-type (IFN gamma(+/+)) mice did not restore granuloma formation, although this treatment diminished IL-10 production in IFN-gamma(-/-) mice. Adoptive transfer of splenocytes from infected IFN-gamma(-/-) mice into infected IFN-gamma(+/+) reduced granuloma formation. These results suggest that splenocytes of IFN-gamma( /-) mice suppress granuloma formation. On the other hand, although IFN-gamma production induced by R. aurantiacus infection was detected in nude mice, which are deficient in T cells, granuloma formation was not induced in them. However, adoptive transfer of immune splenocytes from either IFN-gamma(+/+) mice or IFN gamma(-/-) mice could induce granuloma formation. This means that splenocytes of IFN-gamma(-/-) mice have the ability to both induce and suppress granuloma formation. Induction of granuloma is probably dependent on both T cells and IFN gamma produced by non-T cells. It is suggested that the role of T cells in granuloma formation is not dependent on their IFN-gamma production. PMID- 11478673 TI - Development of systemic bacteraemia after oral inoculation of vancomycin resistant enterococci in mice. AB - Bacteraemia caused by vancomycin-resistant enterococci (VRE) is an important clinical problem because there are only a few potent antimicrobial agents against such bacteria. Therefore, understanding the pathogenic mechanisms of VRE bacteraemia is important for prophylaxis. This study shows that treatment of mice with cyclophosphamide and a combination of metronidazole, kanamycin and vancomycin reduced normal intestinal flora and induced systemic VRE bacteraemia. Translocation of VRE and the normal intestinal flora to the mesenteric lymph nodes, liver, spleen and blood, and mortality rate were dependent on treatment with cyclophosphamide and each of the three antimicrobial drugs. Among the different strains studied, C57BL/6 mice were the most susceptible to VRE. The virulence of vancomycin-resistant Enterococcus faecalis was greater than that of vancomycin-resistant Ent. faecium. On the day after inoculation of VRE, Escherichia coli was also detected in many VRE-positive specimens including blood, liver and the mesenteric lymph nodes. Moreover, both VRE and E. coli were detected simultaneously in almost all blood samples obtained from dead and dying mice, and VRE organisms outnumbered E. coli in those samples by 100:1 or more. These results indicate that changes in normal intestinal flora by administration of antimicrobial drugs and severity of neutropenia induced by cyclophosphamide are important factors that contribute to the development of systemic VRE bacteraemia. E. coli may be intimately associated with the establishment of VRE translocation. PMID- 11478674 TI - Susceptibility of irradiated mice to Bacillus anthracis sterne by the intratracheal route of infection. AB - The susceptibility of sublethally irradiated mice to pulmonary infection with Bacillus anthracis was investigated in a mouse model. Female B6D2F1/J mice were challenged intratracheally with 4.3 x 10(6), 3.7 x 10(7) and 4.4 x 10(8) cfu of B. anthracis Sterne spores 4 days after 60Co gamma-radiation at a dose of 0, 1, 2, 3, 4, 5, 6 or 7 Gy. Bacterial cultures were obtained from lung, spleen homogenates and heart blood. A biphasic mode of mortality was observed, with a constant response of up to 3 or 4 Gy (up to 18% mortality), after which a sharp increase in mortality occurred (up to 100%). When irradiation was delayed beyond 15 days after inoculation, the susceptibility to B. anthracis infection and subsequent mortality disappeared. B. anthracis was recovered from the organs and blood of up to 89% of the animals. However, organisms of enteric origin were also isolated mixed with B. anthracis from up to 36% of the animals exposed to 3, 5 or 7 Gy. Inoculation of B. anthracis delta-Sterne-1 that lacks lethal toxin and oedema toxin also induced infection with B. anthracis, but not translocation of enteric micro-organisms. The synergic adverse effect of exposure to gamma radiation followed by intratracheal challenge with B. anthracis was observed above 4 Gy. The lethal toxin of B. anthracis may enhance the emergence of polymicrobial infection with B. anthracis and enteric micro-organisms. PMID- 11478675 TI - The N-terminal of thrombospondin-1 is essential for coagulase-negative staphylococcal binding. AB - Bacterial binding was studied to determine whether thrombospondin-1 (TSP) acts as a ligand in attachment of coagulase-negative staphylococci (CNS). Twenty-five of 27 CNS strains bound soluble TSP. Staphylococcus epidermidis J9P bound 125I labelled TSP in a dose-dependent manner. Scatchard plot analysis of the binding of TSP by strain J9P revealed two Kd values of 6.4 x 10(-9) M and 2.9 x 10(-8) M. The binding structures of strain J9P were sensitive to protease and were resistant to heat treatment. Unlabelled TSP and recombinant von Willebrand factor inhibited binding of TSP by strain J9P, but other proteins or monosaccharides did not. Heparin inhibited binding of TSP to strain J9P and two other S. epidermidis strains, BD5703 and BD969. Fusion proteins of the type 1 repeats, type 2 repeats, type 3 repeats and C-terminal domain of TSP or the synthetic Arg-Gly-Asp peptide did not inhibit binding of TSP to bacteria. TSP promoted adhesion of S. epidermidis strains when it was immobilised on polymer surfaces. These results indicate that the specific interaction between CNS and TSP may contribute to bacterial adhesion on biomaterial surfaces. The N-terminal heparin-binding domain of TSP appears to be the major region for recognition by CNS. PMID- 11478676 TI - Colonisation and transmission of Clostridium difficile in healthy individuals examined by PCR ribotyping and pulsed-field gel electrophoresis. AB - Healthy adults who had not been exposed to antimicrobial agents for the preceding 4 weeks were examined for intestinal carriage of Clostridium difficile. The 1234 individuals examined were composed of seven groups: three classes of university students, hospital workers at two hospitals, employees of a company and self defence force personnel at a local station. Overall, 94 (7.6%) individuals were positive for C. difficile by faecal culture but carriage rates among the study groups ranged from 4.2% to 15.3%. Typing by PCR ribotyping and pulsed-field gel electrophoresis demonstrated clusters of carriers colonised by a single type in each of three groups, indicating that cross-transmission of C. difficile can occur in community settings. Follow-up culture was performed on 38 C. difficile positive individuals and C. difficile was isolated again from 12 (32%) of them 5 7 months after the initial culture; six (50%) of these 12 individuals had a new strain on repeat culture. Two or more family members were C. difficile-positive in five of 22 families examined. C. difficile with an identical type was isolated from persons within a family in only one family. These results suggest that intestinal carriage by healthy adults may play a role as a reservoir for community-acquired C. difficile-associated diarrhoea, but that cross-transmission of C. difficile does not occur frequently among family members at home. PMID- 11478677 TI - Flagellin gene PCR-RFLP analysis of a panel of strains from the Burkholderia cepacia complex. AB - Burkholderia cepacia, an important opportunist pathogen, is genetically heterogeneous. The B. cepacia complex has been subdivided into a number of genospecies or genomovars. A flagellin gene PCR-RFLP method was applied to a representative panel of strains of known genomovar. The technique was able to distinguish strains of B. multivorans from other members of the B. cepacia complex on the basis of amplicon size (typical of type I rather than type II flagellins) with the exception of one genomovar I strain. There was considerable variation in RFLP patterns amongst the panel of strains; only two pairs of strains were indistinguishable with both HaeIII and MspI digestion. Where RFLP patterns matched with both enzymes or a single enzyme, matching strains were always in the same genomovar. It was possible to distinguish the UK cystic fibrosis epidemic strain from all other members of the panel, including nine other genomovar III strains. The level of variation suggests that flagellin genotyping is a useful method for discriminating between B. cepacia strains. PMID- 11478678 TI - Analysis of different molecular methods for typing methicillin-resistant Staphylococcus aureus isolates belonging to the Brazilian epidemic clone. AB - The extensive geographic spread of MRSA isolates belonging to the Brazilian epidemic clone (BEC) limited the value of pulsed-field gel electrophoresis (PFGE) in epidemiological studies of outbreaks caused by these strains. Thus, the discriminatory power of eight different molecular methods was evaluated in an attempt to establish a methodology for genotyping BEC isolates involved in intra hospital outbreaks. BEC isolates from five hospitals in Teresina City, Piaui State were genotyped by conventional electrophoresis or PFGE of Cla I- or Sma I digested genomic DNA hybridised with specific labelled mecA, Tn554, IS257 and IS256 probes. The combination of PFGE with Cla I/mecA, Cla I/Tn554, Cla I/IS257, Sma I/mecA and Sma I/IS257 probe-fingerprinting techniques provided a very poor discriminatory power for BEC strains. Although Cla I/IS256 fingerprinting discriminated 17 different polymorphisms among the isolates displaying PFGE A1 pattern, this strategy was not reproducible. In contrast, the combination of PFGE and Sma I/IS256 polymorphisms differentiated BEC isolates into nine stable polymorphisms. Thus combination of PFGE and hybridisation with IS256 probe may be recommended as a useful means of typing BEC strains involved in intra-hospital infections. PMID- 11478679 TI - Different isoforms of secreted aspartyl proteinases (Sap) are expressed by Candida albicans during oral and cutaneous candidosis in vivo. AB - Distinct isoforms of secreted aspartyl proteinases (Sap) of Candida albicans are important virulence factors for different types of candidosis. Predominant expression of Sap1-3 has been shown to be crucial for superficial infections in experimental mucosal and cutaneous candidosis, whereas Sap4-6 might be important for systemic disease. This in-vivo study investigated Sap expression in two samples from patients with oral candidosis and from cutaneous infection. Two different polyclonal antibodies directed against Sap1-3 and Sap4-6 were used for ultrastructural characterisation of protein localisation and expression. Post embedding immuno-electron microscopy revealed Sap1-3 and Sap4-6 immunoreactivity in all samples. All C. albicans cells expressed predominantly the proteinases Sap1-3 which were evenly distributed within the cell wall and cytoplasmic membrane. In contrast, Sap4-6 labelling was only evident in a few fungal cells. In particular it was localised at the tips of hyphal cells during invasion. These data suggest a different pathogenetic role for Sap1-3 and Sap4-6 during host fungal interaction. PMID- 11478680 TI - Epidemiology of breast cancer: an environmental disease? AB - Breast cancer is the leading cancer site in women, both in the developed and the developing world. Incidence rates are increasing in many countries, although, in some, mortality may be stable or slightly decreasing. Geographical differences exist, with high rates of disease in North America, North Europe and Oceania, intermediate rates in South and Central America as well as South and East Europe, and low rates in Africa and Asia. Most of the literature reports that genetic inherited factors account for less than 5% of cases, although some authors advance higher figures, up to about 10%. Risk factors for breast cancer are related to the reproductive life of women: early menarche, nulliparity or late age at first birth, late menopause, diet and physical exercise, as well as hormonal factors, be they endogenous (high levels of free or not bound to SHBG estrogens) or exogenous (long-term use of oral contraceptives or menopausal hormone replacement). The present review does not aim to be exhaustive and fully comprehensive, or to present in detail domains currently well known and accepted by all. On the contrary, it modestly wishes to highlight potentially controversial conditions which could in the future be recognized as new risk factors. PMID- 11478681 TI - Semen quality and male reproductive health: the controversy about human sperm concentration decline. AB - Concern about the effect of environmental changes on male reproductive health has grown in recent years to become a major preoccupation in some developed countries. A possible decline in human sperm concentration was suggested in the early seventies following studies in the US. In 1992 a meta-analysis of 61 articles published by Carlsen et al. concluded that the mean sperm count of healthy men had declined by 1% per year over the previous 50 years. From 1995 and onwards, some retrospective, longitudinal analyses of the sperm count of fertile or infertile men contradicted this while others did not. The demonstration of a geographical variation in sperm concentration, between and within countries or regions, appears to be less controversial. The amplitude of the difference observed cannot only be explained by methodological or confounding factors, and must to some extent be attributed to ethnic, genetic or environmental factors. As many of the published studies suffer from imprecision regarding the description of population characteristics and confounding factors, and were not designed with controlled and standardised methodology, the debate remains open. Prospective studies in well-defined cohorts of men in various populations are required to evaluate the potential effect of external factors on male reproductive health. These studies should not be limited to the analysis of sperm concentration, as this may not be the best biomarker of testis function and human fertility. PMID- 11478682 TI - Tissue-specific expression pattern of estrogen receptors (ER): quantification of ER alpha and ER beta mRNA with real-time RT-PCR. AB - We have examined the tissue-specific mRNA expression of ER alpha and ER beta in various bovine tissues using real-time RT-PCR. The goal of this study was to evaluate the deviating tissue sensitivities and the influence of the estrogenic active preparation RALGRO on the tissue-specific expression and regulation of both ER subtypes. RALGRO contains Zeranol (alpha-Zearalanol), a derivative of the mycotoxin Zearalenon, shows strong estrogenic and anabolic effects, and exhibits all symptoms of hyperestrogenism, in particular reproductive and developmental disorders. Eight heifers were treated over 8 weeks with multiple-dose implantations (0x, 1x, 3x, 10x) of Zeranol. Plasma Zeranol concentration, measured by enzyme immunoassay, of multiple treated heifers was elevated. To quantify ER alpha and ER beta transcripts also in low-abundant tissues, sensitive and reliable real-time RT-PCR quantification methods were developed and validated on the LightCycler. Expression results indicate the existence of both ER subtypes in all 15 investigated tissues. All tissues exhibited a specific ER alpha and ER beta expression pattern and regulation. With increasing Zeranol concentrations, a significant downregulation of ER alpha mRNA expression could be observed in jejunum (p<0.001) and kidney medulla (p<0.05). These data support the hypothesis that ER beta may have different biological functions than ER alpha, especially in kidney and jejunum. PMID- 11478683 TI - Genetic variation in physiological sensitivity to estrogen in mice. AB - Genetic variation in susceptibility to endocrine disruption by estrogenic agents was examined in juvenile male mice. Mice were implanted with increasing doses of estradiol (E2) at 3 weeks of age and reproductive responses were determined 3 weeks later. Greater than 16-fold differences in susceptibility to the disruption of reproductive development by E2 were detected between strains of mice. CD-1 was much more resistant to the inhibition of testes weight, vesicular gland weight and spermatogenesis by increasing doses of E2. Spermatid maturation was eliminated by low doses of E2 in unselected strains such as C17/Jls and C57BL/6J. In contrast, widely used, large litter size selected CD-1 mice showed little or no inhibition in spermatogenesis even in response to 16-fold higher doses of E2. Testicular sulfotransferase activity (EST) per gram body weight was 3.5-fold higher in untreated CD-1 than in B6 strain males. This suggests that genetic differences in testicular EST activity may play a critical role in the detoxification of estrogens. These and other findings emphasize the need to identify and study genetic variation in sensitivity to estrogen in laboratory animal models used to assess the risk of xenobiotic estrogen exposure. PMID- 11478684 TI - Uterotrophic effect of a saturated fatty acid 17-ester of estradiol-17beta administered orally to juvenile rats. AB - In comparison to estradiol-17beta, the naturally synthesized estradiol-17beta-17 fatty acid esters are potent estrogens when administered subcutaneously. A lipophilic character of estradiol-17-esters could partially protect them from metabolic inactivation. In order to compare their relative estrogenic potency when administered orally, the uterotrophic response to different dosages (0, 2.5, 25, 250 and 2500 nmol/kg BW/day) of estradiol-17beta and estradiol-17beta-17 stearate was assessed in juvenile Sprague-Dawley female rats. Estrogens were administered by oral gavage once a day for 6 days. On the 7th day uterus and vagina were dissected, weighed, and examined microscopically. At 2.5 and 25 nmol/kg BW/day, no difference was detected in the uterus weight compared to control animals which received the vehicle alone (corn oil). At 250 nmol/kg BW/day, the uterotrophic response was maximal in estradiol-17beta-17-stearate treated animals (x2.40-2.70), whereas it was moderate in estradiol-17beta-treated rats (x1.86) at the same dosage. This differential weight gain effect of estradiol-17beta-17-stearate was correlated with typical microscopic changes in uterus and vagina. The results are in favour of a stronger estrogenic effect of orally given lipoidal estrogens compared to estradiol-17beta. This could be explained by a slower but sustained absorption of estradiol-17beta released from estradiol-17beta-17-stearate by esterases and/or by a facilitated transfer of esters in the lymphatic circulation. PMID- 11478685 TI - Immunolocalization of intercellular adhesion molecule-1 and leukocyte functional associated antigen-1 in schistosomal soluble egg antigen-induced granulomatous hyporesponsiveness. AB - In this work, the changes in expression of the adhesion molecules ICAM-1/LFA-1 on inflammatory cells of the liver were studied by immunohistochemistry. Mice sensitized with SEA and infected with S. mansoni and S. mansoni-infected controls were examined from day 35 to day 56 postinfection. A significant upregulation of ICAM-1 and LFA-1 in both the SEA group and the infected control group started shortly after egg deposition at day 35 and persisted up to day 56 p.i. Notably, both ICAM-1 and LFA-1 expression peaks were shifted earlier to day 38 p.i. in the SEA group compared to day 40 in the infected control group. The distribution of ICAM-1 and LFA-1 in both groups was comparable. At the early phase of infection before granuloma formation, both ICAM-1 and LFA-1 were detected along the sinusoidal wall of small blood vessels. At the acute cellular granuloma phase, they were homogeneously distributed all over the inflammatory cells, while at the chronic fibrocellular stage a non-homogeneous staining of granuloma cells at the periphery of the granuloma was apparent. The present data suggest that adhesion molecules play a role in the initiation and maintenance of granuloma formation. Thus, the granulomatous hyporesponsiveness induced by sensitization with SEA was associated with reduced expression of adhesion molecules. PMID- 11478686 TI - Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo. AB - Cystic forms (also called spheroplasts or starvation forms) and their ability to reconvert into normal motile spirochetes have already been demonstrated in the Borrelia burgdorferi sensu lato complex. The aim of this study was to determine whether motile B. garinii could develop from cystic forms, not only in vitro but also in vivo, in cyst-inoculated mice. The cysts prepared in distilled water were able to reconvert into normal motile spirochetes at any time during in vitro experiments, lasting one month, even after freeze-thawing of the cysts. Motile spirochetes were successfully isolated from 2 out of 15 mice inoculated intraperitoneally with cystic forms, showing the infectivity of the cysts. The demonstrated capacity of the cysts to reconvert into motile spirochetes in vivo and their surprising resistance to adverse environmental conditions should lead to further studies on the role and function of these forms in Lyme disease. PMID- 11478687 TI - Arbitrarily primed PCR and sequencing of 16S rDNA for epidemiological typing and species identification of Burkholderia cepacia isolates from Swedish patients with cystic fibrosis reveal genetic heterogeneity. AB - To investigate whether arbitrarily primed (AP)-PCR and/or 16S rDNA sequencing could be used as rapid methods for epidemiological typing and species identification of clinical Burkholderia isolates from patients with cystic fibrosis (CF), a total of 39 clinical B. cepacia isolates, including 33 isolates from 14 CF patients, were fingerprinted. ERIC-2 primer was used for AP-PCR. The AP-PCR clustering analysis resulted in 14 different clusters at a 70% similarity level. The AP-PRC patterns were individual despite considerable similarities. To sequence rDNA, a broad-range PCR was applied. The PCR product included four variable loops (V8, V3, V4 and V9) of the 16S ribosomal small subunit RNA gene. The multiple sequence alignment produced 12 different patterns, 5 of them including more than one isolate. Heterogeneity of the bases in the V3 region, indicating the simultaneous presence of at least two different types of 16S rRNA genes in the same cell, was revealed in 10 isolates. Most of the CF patients were adults who had advanced disease at follow-up. Both the sequencing and the AP-PCR patterns revealed genetic heterogeneity of isolates between patients. According to the results obtained, AP-PCR could advantageously be used for epidemiological typing of Burkholderia, whereas partial species identification could effectively be obtained by sequencing of the V3 region of the 16S RNA gene. PMID- 11478688 TI - Study of antioxidant effect of apigenin, luteolin and quercetin by DNA protective method. AB - A DNA protective capacity of three flavonoids, apigenin (AP), luteolin (LU) and quercetin (QU) against free radicals generated by H202, resp. Fe2+ is reported. This effect corresponding with scavenging of free radicals or with chelating of iron was assayed at two concentrations of flavonoids studied (1 microM and 10 microM). The quantitative analysis has shown that LU possesses the highest DNA protective effect of flavonoids investigated in the presence of H2O2. On the other hand, in the presence of 10 microM Fe2+, AP exhibited the highest DNA protective effect at the concentration of 1 microM and the following order was reached at the stoichiometric concentrations (10 microM) of Fe2+. It is believed that this discrepancy is caused by the ability of LU and QU iron-complex formation as it was separately investigated using UV-VIS spectrometry. PMID- 11478689 TI - Arylamine N-acetyltransferase activities in human breast cancer tissues. AB - N-Acetyltransferase activities were determined in tumor (12 malignant and 6 benign) and control (non-cancerous) breast tissues from 18 female patients. The activities of matched 12 malignant tumor and control tissue cytosols showed 6 rapid, 4 intermediate and 2 slow acetylators based on p-aminobenzoic acid (NAT1) and sulfamethazine (NAT2) as substrates. Compared to the activities of slow acetylators, the rapid acetylators exhibited mean apparent Vmax values about 5- and 50-fold greater for p-aminobenzoic acid and sulfamethazine, respectively. No correlation was observed between the blood and breast tissue N-acetyltransferase (NAT1 and NAT2) activities. When the mean apparent N-acetyltransferase activities of the malignant and benign breast tumor tissues were compared, the results showed an increased activity for both p-aminobenzoic acid (PABA) and sulfamethazine (SMZ) acetylation in the malignant tissues compared to benign ones, and also control tissues showed lower activities compared to tumor tissues. Moreover, the mean NAT2 activity was about 2-fold greater in the malignant tissues when compared to NAT1 activity. PMID- 11478690 TI - Expression of cyclin A in human leukemia cell lines HL-60 and K-562 at the level of light and electron microscope by using immunocytochemical methods. AB - In this study subcellular localization of cyclin A in the human promyelocytic leukemia cell line HL-60 and the human erythroleukemic K-562 cell line was examined by immunocytochemical methods. Studies were based on light and electron microscope evaluations. Cyclin A at the level of light microscope was present in 48% of the cells in HL-60 cell line and in 40% of the cells in K-562 line. Streptavidin-gold method was used for localization of cyclin A at the ultrastructural level. There was expression of cyclin A in the nucleus and in the cytoplasm. In the nucleus gold particles were seen to be associated with the condensed chromatin of the both leukemia cell lines. In the cytoplasm cyclin A was concentrated at a low level and was associated with ribosomes. Controls of the leukemia cells incubated with normal mouse serum showed no labeling at the light and electron microscope level. PMID- 11478691 TI - Evaluation of MMP-1, MMP-8, MMP-9 serum levels in patients with adrenal tumors prior to and after surgery. AB - The aim of the study is to evaluate MMP-1, MMP-8 and MMP-9 serum levels in patients with adrenal tumors prior to and after surgery. Metalloproteinase-1 (MMP 1), MMP-8 and MMP-9 serum levels were evaluated in 43 patients operated on at our clinic between 1997-1999. Forty-one (95.3%) patients underwent adrenalectomy. Two (4.7%) patients were disqualified from surgery due to infiltration of adjacent tissues. MMP-1, MMP-8 and MMP-9 serum levels were determined at the admission and in case of surgery again one month after the operation. ELISA assay (K&D) was applied. Tumor type was determined on the basis of clinical, hormonal and histopathological examination. The correlation between MMP levels and tumor sizes was also evaluated. Patients were divided into 6 groups. Group I included 11 patients with adrenocortical carcinoma (4 with Cushing's syndrome and 7 with incidentalomas); group II--6 patients with benign hormonally active adrenocortical adenoma (4 with Cushing's syndrome and 2 with Conn's syndrome); group III--patients with benign, hormonally inactive adenocortical adenoma; group IV--6 patients with benign, hormonally active phaeochromocytoma; group V--4 patients with hormonally inactive phaeochromocytoma; group VI--5 patients with hormonally inactive adrenal tumors of extraglandular origin (2 myolipomas, 2 fibrolipomas, 1 hammartoma). The control group comprised 10 healthy individuals. Increased MMP-8 and MMP-9 levels were noted in patients with benign and malignant adrenal tumors. No increase of MMP levels was found in patients with tumors of extraglandular origin. The increased MMP-8 and MMP-9 levels occurred most frequently in patients with adrenocortical and hormonally active adrenomedullar cancer, and most rarely in patients with hormonally active adrenocortical tumors. MMP-8 and MMP-9 serum levels did not significantly differ between patients with adrenocortical incidentaloma cancers and in patients with benign incidentalomas. MMP-8 and MMP-9 levels were not increased in patients with inoperable adrenocortical cancers. Serum MMP-1 levels were not increased in patients with benign and malignant adrenal tumors. After surgery, MMP-8 and MMP-9 levels decreased significantly in patients with adrenocortical cancers, whereas the decrease of these MMPs in patients with benign tumors, although noticeable, was not statistically significant. MMP-8 and MMP-9 levels decreased significantly in all patients with increased preoperative levels, although they remained higher than the maximum normal values only in few patients (in 7 and 2 patients, respectively). No correlation between the levels of evaluated MMPs and tumor sizes were found. PMID- 11478692 TI - Location and incidence of chromosome and chromatid breaks in patients with Hodgkin's disease or testicular tumors. AB - In 90 patients aged 17 to 35 who suffered from Hodgkin's disease (HD) or had testicular tumors (TT), the location of chromosome and chromatid breaks on individual chromosome segments was reviewed using an adapted Funes-Cravioto scheme, in addition to examining the percentage of structural chromosomal aberrations. On the basis of an analysis of 1121 breaks in patients with HD or TT, the results were presented graphically as multiples of the expected number of breaks for the normal population. Before the beginning of treatment, the number of structural chromosomal aberrations (SCA) in patients with TT or HD was equal to that in a control group of subjects with malignant diseases. This, however, does not apply to the location of chromosome and chromatid breaks. In patients with HD, the dominant unstable sites are located on group A2 chromosomes, segments 2 and 5, and on group B chromosomes, segment 5. In patients with TT, the number of chromosome and chromatid breaks is also increased on group A2 chromosomes, segments 2 and 5, and in addition, also on group B chromosomes, segment 4. PMID- 11478693 TI - Polymer conjugated bovine seminal ribonuclease inhibits growth of solid tumors and development of metastases in mice. AB - Bovine seminal ribonuclease (BS-RNase) exerts a potent cytotoxic activity when administered intratumorally (i.t.) to the nude mice bearing human tumors. The ineffective treatment with intravenous (i.v.) or intraperitoneal (i.p.) administration led us to the synthesis of polymeric conjugates with BS-RNase to prevent it from degradation in the blood vessel. Hydrophilic poly[N-(2 hydroxypropyl)methacrylamide] (PHPMA) was used for BS-RNase modification and a PHPMA-BS-RNase conjugates were prepared. Classic conjugate (P-BS) with BS-RNase bound to the polymer by its oligopeptide site chains was prepared by aminolytic reaction of the polymer precursor bearing reactive ester groups situated in the side chains of polymer, while star-like conjugate (S-BS) was synthesized by the reaction of PHPMA containing end-chain reactive group with BS-RNase in aqueous buffer solution at pH 8. In contrast to the total ineffectiveness of free BS RNase administered i.v. at a daily dose 10 mg/kg, application of P-BS and S-BS conjugates at doses 2 mg/kg and 0.5 mg/kg caused significant inhibition of the growth of human melanoma in nude mice. On the base of these results the effect of i.v. administered S-BS on the metastatic process and the survival of C57Bl/6 inbred mice inoculated with B16 melanoma cells was investigated. Sixty per cent of mice treated with S-BS (0.5 mg/kg/day) survived 100 days without metastatic foci when the experiment terminated. The average survival time of the treated groups was 75.5 days compared to 32.7 days in the control group. BS-RNase conjugated to water soluble polymers appears to be the first BS RNase preparation which exerts anticancer and antimetastatic activity following its intravenous administration. PMID- 11478694 TI - Effects of short term treatment with pentagastrin, proglumide, tamoxifen given separately or together with 5-fluorouracil on the growth in the murine transplantable Colon 38 cancer. AB - It is well known that 5-fluorouracil (5-FU) is the most effective drug in the treatment of colon cancer, however the positive response is small, only about 20%. On the other hand, it has been postulated that the growth of colon cancer depends on many growth factors, such as gastrin and estrogens. The search for new substances increasing the antitumor effect of 5-FU has lasted for many years. The aim of the present study was to examine the effects of pentagastrin (PEN, syntetic gastrin analogue), proglumide (PRO, a blocker of gastrin receptor) and tamoxifen (TAM, a partial estrogen antagonist) given separately or together with 5-FU on proliferation, apoptosis, necrosis and proliferation/apoptosis (P/A) ratio in the murine transplantable Colon 38 cancer. The male mice were implanted with a suspension of Colon 38 cells. After 7 days, the animals were treated with PEN (250 microg/kg b.w., twice daily), PRO (100 mg/kg. b.w., twice daily), TAM (10 microg/animal) separately or together with 5-FU (60 mg/kg b.w., once) for 2 days. The incorporation of bromodeoxyuridine (BrdU) into cell nuclei was used as an index of cell proliferation (labeling index--LI). The in situ labeling of nuclear DNA fragmentation according to TUNEL method was considered as an apoptotic index (AI). It was found that 5-FU increased the apoptosis, unexpectedly increased the LI and decreased the P/A ratio when compared to control. Both PEN and PRO increased the apoptosis and in the case of PRO decreased P/A ratio when compared to control. TAM did not affect any of the examined parameters. All of the investigated substances modify the 5-FU action: PEN and PRO on AI and LI and TAM on AI and P/A ratio. Necrosis was observed in 3 tumors treated with PEN + 5-FU, in 2 tumors of PRO + 5-FU group and in 1 tumor of group with 5-FU and with PEN. Further studies are needed to elucidate if those modification of 5-FU action by the examined substances will be useful in the inhibition of the growth of Colon 38 cancer. PMID- 11478695 TI - Further studies of blood levels of some tumor markers in the area polluted by polychlorinated biphenyls and control population. AB - The levels of beta2-microglobulin (beta2-m), alpha-fetoprotein (AFP) and thyroglobulin (TG) were measured in the serum of 245 employees of chemical factory formerly producing polychlorinated biphenyls (PCB) consisting of 54 males (age range 24-65 years, median 45) and 191 females (age range 20-69 years, median 45). The control population consisted of 636 adults from control areas of northwest and east Slovakia. The frequency of beta2-microglobulin levels lower than 1.6 microg/ml in 242 employees of chemical factory was 76.8% (186/242) which was three times higher (P<0.001) than 24.4% (155/635) in 636 controls. Still more remarkable difference was obtained when using the cut/off level of 1.2 microg/ml, the frequency of such values in the employees being 45.4% (110/242) vs. 4.4% (28/635) in the controls. In contrast, no difference in alpha-fetoprotein levels was observed between the employees and the controls, the respective frequency of these < 5.0 ng/ml being 87.6% (212/242) vs. 86.2% (389/451) and these < 10.0 ng/ml being 100.0% (242/242) vs. 97.8% (441/451). Similarly, the frequency of normal thyroglobulin levels < 50.0 ng/ml) did not differ, being 95.6% (174/182) in the employees and 87.9% (87/99) in the controls. Most of a total of 20 cases with thyroglobulin level > 50.0 ng/ml showed sonographicaly enlarged and multinodular thyroid with focal or diffuse hypoechogenicity, three of them showed solitary nodule with a diameter > 10 mm. Although the decreased levels of beta2 microglobulin might be somehow related to the modulation of immune system, more plausible explanation appears to be the possible impairment of renal tubules by PCB similar to that caused by heavy metals resulting in increased urinary excretion of beta2-microglobulin and decrease of its blood level. PMID- 11478696 TI - Micronuclei frequencies in exfoliated nasal mucosa cells from pathology and anatomy laboratory workers exposed to formaldehyde. AB - Formaldehyde (FA) is a widely used industrial chemical. Sufficient evidence exists to consider FA as an animal carcinogen. A possible causal role for FA may be considered likely for cancer of the nasopharynx and the nasal cavities in humans. The frequency of micronuclei (MN) in cells of the nasal mucosa was evaluated for 23 individuals in pathology and anatomy laboratories exposed to FA. Twenty-five healthy subjects were selected from the university and hospital staff as a control group. The measured air concentrations of FA in the breathing zone of the laboratory workers were between 2 and 4 ppm. The mean +/- SD values of nasal mucosa MN (per 1000) frequency from exposed and controls were 1.01 +/- 0.62 and 0.61 +/- 0.27, respectively (p < 0.01). Effect of smoking, age, sex and duration of exposure on the genotoxicity parameters analyzed were also evaluated. Our data suggest that low level exposure to FA is associated with cytogenetic changes in epithelial cells of the nasal region and that nasal mucosa cells exposed through respiration is an important target of FA-induced genotoxic effects. PMID- 11478697 TI - High dose rate intraluminal brachytherapy in the treatment of malignant airway obstructions. AB - Endobronchial brachytherapy has been increasingly used in an effort to improve local control and relieve symptoms of malignant airway obstructions. Results of the high dose rate (HDR) intraluminal brachytherapy in 67 patients with inoperable endobronchial tumor treated by combination of teletherapy and brachytherapy with curative (group A ) or palliative (group B) intent, patients with recurrent tumors after previous radiotherapy treated by endobronchial brachytherapy alone (group C), and patients treated by brachytherapy without teletherapy (group D) are presented. Symptomatic improvement was achieved in 66%, 74%, 64% and bronchoscopic response in 70%, 85%, 78% of patients in groups A, B and C, respectively. Median survival was 365, 242 and 884 days from diagnosis and 245, 151 and 153 days from the first brachytherapy application in groups A, B and C, respectively. In group D complete bronchoscopic response was achieved in 3 of 4 patients with early tumor and partial response in 6 of 7 patients with advanced disease. We observed 4 acute and 9 late complications. Brachytherapy is an effective palliative treatment of malignant airway stenosis, but the effect on survival is not apparent. PMID- 11478698 TI - Staging in untreated patients with small cell lung cancer. AB - In order to describe the real biological behavior of the small-cell lung cancer we have analyzed survival rates of 66 patients with small-cell lung cancer who did not receive any specific anti cancer therapy. Also, objective of this study was to evaluate the staging system of the small-cell lung cancer. Untreated small cell lung cancer patients with limited stage disease had statistically significant (p < 0.05) better survival rates in comparison to patients with extensive stage disease. T and N factor of the TNM classification did not influence the survival in untreated small-cell lung cancer patients. It appears that the TNM staging system is not predicting survival probabilities of untreated patients with small-cell lung cancer, while the two-stage system appeared very well based on survival probabilities of these patients. PMID- 11478699 TI - Repair of oxidative DNA damage--an important factor reducing cancer risk. Minireview. AB - Oxygen free radicals formed during normal aerobic cellular metabolism generate a variety of DNA lesions including modified bases, abasic sites and single strand breaks with blocked 3' termini. If left unrepaired, these damages may contribute to a number of degenerative processes, including cancer and aging. In most organisms, the repair of oxidative DNA lesions is supposed to be handled by the base excision repair (BER) pathway. BER is a multistep process that involves the sequential activity of several proteins, many of them were isolated and functionally characterized using the simple prokaryotic and lower eukaryotic model systems, Escherichia coli and Saccharomyces cerevisiae, respectively. As the amino acid sequence of DNA repair proteins is often well conserved from bacteria to man, our understanding of BER in higher eukaryotes drives extensively from the microbial models, namely from the yeast S. cerevisiae. Thus, results obtained on a simple yeast model are a source of new information, which can be used as a paradigm for all eukaryotic cells. PMID- 11478700 TI - Evaluation of p53 and bcl-2 oncoprotein expression in precancerous lesions of the oral cavity. AB - The oral cavity is continually exposed to various traumas due to the effect of thermal, mechanical and chemical stimuli, which when accompanied by inflammatory states may promote the growth of neoplastic changes. Numerous studies have revealed a correlation between the expression of p53 and Bcl-2 proteins and the progression of neoplastic disease. It cannot be excluded that these proteins act as biomarkers of a neoplastic transformation threatening in precancerous states (including leukoplakia) or the already existing neoplastic transformation (e.g. in oral squamous cell carcinoma). The aim of the study was to evaluate the expression of p53 and Bcl-2 proteins in the proliferating epithelium in relation to leukoplakia degree and with regard to the lesions accompanied and not accompanied by squamous cell carcinomas. Fifty-five cases of proliferating changes in the oral epithelium (leukoplakia) were investigated. Group I contained 20 leukoplakias not accompanied by oral squamous cell carcinomas. Groups II, III and IV included 35 cases of changes in the vicinity of carcinomas on the lower lip (group II), in the front 2/3 of the tongue (group III) and in the oral floor (group IV). Staining was performed according to the immunohistochemical method with the use of monoclonal antibodies against human p53 protein (DAKO No M7001) and Bcl-2 (DAKO No M0887). A higher expression of p53 protein (54%) was found in leukoplakia changes coexisting with squamous cell carcinomas, compared with the non-accompanied ones (p53--45%). The results indicate a correlation between epithelial dysplasia degree and p53 and Bcl-2 protein expression--severe dysplasia occurred with an increase in the expression of both proteins. Leukoplakias situated in the vicinity of squamous cell carcinomas showed higher expression of p53 and Bcl-2 compared with the non-accompanied alterations. A correlation was also revealed between the location and p53 and Bcl-2 protein expression degree in the non-accompanied changes; no such correlations were found in proliferating epithelial changes adjacent to neoplastic tumors. PMID- 11478701 TI - Induction of heat shock protein 70 in drug-resistant cells by anticancer drugs and hyperthermia. AB - The altered constitutive and inducible levels of heat shock proteins 70 (Hsp70) in drug-resistant cells may influence the efficiency of combined hyperthermia and anticancer drug treatment. In the present study, the constitutive levels of Hsp70 and induction of these proteins by hyperthermia and two anticancer drugs (used for resistance development) were determined in cervical and laryngeal carcinoma cells. The levels of Hsp70 were quantified by Western blot. Constitutive levels of Hsp70 were similar in parental and drug-resistant cells suggesting that Hsp70 is not involved in drug-resistance. Hyperthermic treatment induced Hsp70 in all examined cell lines but with different kinetics between drug-resistant and parental cells. Following the treatment with anticancer drugs, Hsp70 was induced only in cisplatin-resistant laryngeal cells. Kinetics of Hsp70 induction (stress type and cell-type specific) was different in drug-resistant cells as compared to parental cells. The observed alterations in Hsp70 induction in drug resistant and parental cells should be taken into account when combined treatments (i. e. hyperthermia and anticancer drugs) are planned. PMID- 11478730 TI - Whose life task is it anyway? Social appraisal and life task pursuit. AB - The study uses a social contextual framework to examine how others are represented in individuals' life task appraisals and how such appraisals are related to strategies to pursue those life tasks. The extent to which life tasks nominated by 81 college students (45 females) were appraised as shared with others and pursued collaboratively was examined through a questionnaire. Participants listed five life tasks, indicated if each life task was theirs alone, indirectly shared (others were impacted by their life task pursuit), or directly shared (another person might also nominate the life task as theirs) with others in their lives, described three activities used to pursue their life tasks, and coded how others were involved inthese activities. Results indicated that the majority of college students' life tasks were appraised as involving others, that directly shared appraisals were found more frequently in the relationship domain, and that directly shared appraisals were associated with collaborative life task pursuit strategies. In addition, appraising life tasks as directly shared did not appear to reflect aspects of anxiety or compensation. A content analysis of the strategies coded as involving other individuals revealed diverse ways in which others are involved in life task pursuit ranging from active engagement of others to cognitive strategies where others are implied. The findings have implications for research on life tasks and other goal structures in that the structures may involve the social context in adaptive ways. PMID- 11478731 TI - Motive-related memories: content, structure, and affect. AB - Two studies tested hypotheses on the content and structure of autobiographical memories and the affect linked to them. In Study 1, agentic- and communal motivated individuals recorded their most memorable experiences and completed the PANAS each day for 6 weeks. Memories were coded for content and structure. Agentics and communals reported more motive congruent memories, and their congruent memories were structured using more differentiation and integration, respectively. In addition, agentics had slightly higher PA and lower NA scores. In Study 2, agentics and communals recalled an event pertaining to either social separation or connection and then completed an affect measure of agentic and communal items. Agentics recalled more agentic memories in the separation condition and communals recalled more communal memories in the connection condition. Complexity analyses showed that agentics and communals used differentiation and integration respectively to recall their motive-congruent memories. The affect data showed a modestpredicted pattern. Results suggest that implicit motives have an impact on autobiographical memory but are not as clearly related to self-report affect measures, possibly due to method variance. PMID- 11478732 TI - Suppression, repressive-defensiveness, restraint, and distress in metastatic breast cancer: separable or inseparable constructs? AB - A longstanding hypothesis links affective and behavioral inhibition with cancer incidence and progression though it does not clarify psychometric distinctions among related constructs. We hypothesized that repressive-defensiveness, suppression, restraint, and distress would be separable factors in our sample of metastatic breast cancer patients. Our results support the discriminant validity of these constructs in our total sample, and the stability over 1 year in our control group. Using factor analysis, we found 4 separate factors at our prerandomization baseline corresponding closely to hypothesized constructs. Additionally, associations in a multi-trait, multi-occasion (baseline and 1 year) matrix met each of the 3 Campbell and Fiske (1959) criteria of convergent and discriminant validity. Future research testing the links between psychological, physiological, and survival outcomes with affective inhibition in cancer patients will be clearer when informed by these distinctions. PMID- 11478733 TI - Doing and being well (for the most part): adaptive patterns of narrative self evaluation during bereavement. AB - Narrative self-evaluation patterns were studied in relation to longitudinal measures of adaptation to the death of a spouse in midlife. Narrative self evaluations, identified in open-ended interview transcripts at 6 months post loss, were coded as either positive or negative and as either doing-based (evaluations of "what one does") or being-based (evaluations of "what one is"). These narrative variables were then compared with separate, clinical-interview measures of grief at 6, 14, and 25 months post-loss. Results confirmed 3 predictions. First, participants who made an optimal proportion of positive to negative self-evaluations (approximately a 5:1 positive-to-negative ratio) had lower grief levels over time than did those who made either higher or lower proportions. Second, the tendency to focus on evaluations of what one does rather than what one is predicted lower grief levels over time. Third, participants who directly integrated doing-based and being-based self-evaluations had lower grief levels over time than those who did not link the 2 evaluations. Implications for the narrative construction of personal meaning and identity in relation to adaptation are discussed. PMID- 11478734 TI - Alexithymia, emotion, and somatic complaints. AB - Alexithymia, by definition, involves difficulties in identifying and describing emotions and has been assumed to be associated with somatization (i.e., a tendency to express psychological distress in somatic rather than emotional form). Empirical research so far, however, has produced no convincing evidence that alexithymia is more associated with somatic complaints than with emotional complaints or that alexithymia correlates with somatic complaints when negative affect is controlled for. In the present study, alexithymia, as measured by the TAS-20, showed no association with somatic complaints in a community sample of 137 individuals when trait anxiety and depression were controlled. Alexithymia did correlate negatively with positive affect, and positively with negative affect. The former association, however, was much more robust, whereas the latter association was found mainly on subjective trait measures of negative affect (as distinct from state measures and more objective trait measures derived from daily recordings during an 8-week period). It is suggested that the association between alexithymia and lack of positive affect deserves more attention in future research. PMID- 11478735 TI - 5-Aminolevulinic acid inhibits [3H]muscimol binding to human and rat brain synaptic membranes. AB - The interaction of 5-aminolevulinic acid (ALA) with GABA(A) receptors has been proposed to underlie the neurological dysfunctions of ALA-accumulating disorders, such as acute intermittent porphyria. The effects of ALA on [3H]muscimol binding to human and rat cerebral cortical membranes were compared. ALA (0.1-10 mM) significantly inhibited the binding of [3H]muscimol (12 nM), with a similar potency in rat and human membranes (IC50 = 199 vs. 228 microM, respectively). Kinetical analysis revealed that ALA (1 mM) significantly increased the Kd and decreased the Bmax of [3H]muscimol to both rat (100 and 50%, respectively) and human (200 and 40%, respectively) membranes, indicating a mixed-type inhibition. The similarity in the potency and mechanism of the ALA-induced inhibition of muscimol binding in rat and human membranes indicate that rat studies are useful to evaluate the neurotoxic properties of ALA towards the human GABAergic system, and may help to understand the pathophysiology of porphyria. PMID- 11478736 TI - Tumor necrosis factor alpha activates the phosphorylation of ERK, SAPK/JNK, and P38 kinase in primary cultures of neurons. AB - Emerging data indicate that the inflammatory cytokine TNFalpha exerts a neuroprotective effect against brain injury. To better understand the mechanism of action of TNFalpha on neurons we have investigated the possible activation of various MAP kinases. Exposure of neurons to TNFalpha triggered the rapid phosphorylation of three members of the MAP kinase family, i.e., extracellular signal-regulated kinase (ERK1/2), stress-activated protein kinase/JUN N-terminal kinase (SAPK/JNK) and the p38 kinase; this activation occured with the same time course and was transient. The TNFalpha-induced activation of ERK1/2 was specifically prevented by compound PD 98059 a specific inhibitor of the MAP kinase kinase MEK1/2. Activation of ERK1/2 was also specifically inhibited by the xanthogenic derivative D609, a specific inhibitor of phosphoinositide phospholipase C suggesting that TNFalpha signaling in neurons involved the acidic sphingomyelinase. PMID- 11478737 TI - Differential protection and recovery of 5-HT1A receptors from N-ethoxycarbonyl-2 ethoxy-1,2-dihydroquinoline (EEDQ) inactivation in regions of rat brain. AB - The effect of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) on 5-HT1A receptors was studied in Sprague Dawley rats. A single dose of EEDQ (4 mg/kg body wt., i.p.) significantly inactivated 5-HT1A receptors, as measured by [3H]8 hydroxy-2-[di-n-propylamino]-tetralin ([3H]8-OH-DPAT), in cortex (64%, p < 0.0001) and hippocampus (48%, p < 0.0001). A significant (p < 0.01) increase in the affinity of 5-HT1A receptors for radioligand was observed in both regions. A dose dependent protection of cortical 5-HT1A receptors from EEDQ inactivation with pre-treatment of different doses of 8-OH-DPAT (4-20 mg/kg) was observed, along with recovery of affinity of [3H]8-OH-DPAT for 5-HT1A receptors in both regions. Although, a dose of 4 mg/kg of 8-OH-DPAT failed to attenuate the effect of EEDQ on hippocampal 5-HT1A receptors, a significant protection of these receptors was observed with 10 and 20 mg/kg of 8-OH-DPAT. Displacement studies revealed that EEDQ has more affinity for cortical (Ki = 101.3 +/- 11.8 nM) than hippocampal (Ki = 133.5 +/- 25.8 nM) 5-HT1A receptors. A time dependent natural recovery of 5-HT1A receptors from inactivation by a single dose of EEDQ (4 mg/kg) was observed more in cortex compared to hippocampus over a period from 1 day to 14 days. The results of this study suggest that 8-OH-DPAT inhibited EEDQ inactivation of cortical and hippocampal 5-HT1A receptors in a concentration dependent manner. The synthesis and turnover of 5-HT1A receptors differ in cortex and hippocampus, as evident by earlier recovery in the cortex. PMID- 11478738 TI - Acid hydrolase activity of cultured bovine oligodendrocytes. AB - We measured the activity of several acid hydrolases of cultured oligodendrocytes prepared from adult bovine brain white matter to clarify the biochemical basis of bovine oligodendrocytes in vitro. Lysosomal enzyme activities were assayed by using 4-methylumbelliferyl glycosides as substrates. Lysosomal enzyme activities became higher at 8-11 days in vitro (DIV) than 4 DIV. The enrichment in acid hydrolase specific activities in oligodendrocytes may be associated with lysosomal origin of myelin-like membranes. PMID- 11478739 TI - Th1 cytokines stimulate RANTES chemokine secretion by human astroglial cells depending on de novo transcription. AB - Beta-chemokines induce the directional migration of monocytes and T lymphocytes that are implicated in the pathogenesis of multiple sclerosis (MS) lesions. RANTES is a member of the beta-chemokine family that has been detected in the lesions of MS patients. However, the cellular sources of RANTES message and the molecular basis for the regulation of its production in MS lesions are not well understood. Glial cells may be a major source of RANTES in vivo and have been shown to produce RANTES in vitro. Thus, the objective of this study was to establish a model system for studying the regulation of RANTES expression by cytokines in cultured human glial cells, and to determine the mechanism involved in the process. We show that the Th1 cytokines TNF-alpha and IL-1beta independently induce RANTES mRNA and chemokine levels in human U-251 MG astroglial cells, and that these effects are time- and concentration-dependent. In addition, we demonstrate that both cytokines increased the rate of transcription of the RANTES gene, as estimated by in vitro nuclear transcript elongation assays. The transcriptional activity in TNF-alpha-treated U-251 MG cells started to increase at 2 h and peaked at 8 h, with levels more than 14 times greater than controls. We further show that NF-kappaB may play a critical role in the up-regulation of human RANTES gene expression in this system. Gel shift assays revealed an induction of in vitro nuclear extract binding activity to the NF-kappaB element of RANTES in cells incubated with the Th1 cytokines. These observations suggest that human astroglia, within diseased brain, may be stimulated to produce RANTES chemokine in response to TNF-alpha and IL-1beta, and that this effect of the Th1 cytokines is attributed to increase of transcription. PMID- 11478740 TI - Characterization of endogenous amino acid efflux from hippocampal slices during chemically-induced ischemia. AB - Using sodium (NaN3)-induced anoxia plus aglycaemia as a model of chemically induced ischemia, we have characterized the endogenous release of excitatory and inhibitory amino acids from superfused hippocampal slices. Chemical ischemia produced an azide (1-30 mM) dose-dependent increase in the efflux of glutamate, aspartate and GABA. These increases were attenuated to varying degrees by removal of Ca2+, or the addition of the voltage dependent Na+-channel blocker tetrodotoxin (TTX), the selective Ca2+ channel blockers conotoxin MVIIA, MVIIC, and nifedipine, the NMDA antagonist MK801, the AMPA antagonist GYKI-52466. Similarly, addition of the GLT-1 glutamate transport inhibitor dihydrokainate (DHK) and the anti-estrogen/anion channel blocker tamoxifen also attenuated the efflux of glutamate and GABA. It would therefore appear that the increases in amino acid efflux induced by chemical ischemia originates from both the neuronal pool, via conventional exocytotic release, and glial sources via reversal of the GLT-1 transporter and anion channel regulated cell swelling. PMID- 11478741 TI - Age-associated hypothalamic glutamate receptor density: effect of dietary protein. AB - An age-associated (3-18 months) increase in hypothalamic glutamate receptor (HgluR) binding due to increase in receptor density (Bmax) was observed in rats maintained with normal (20%) protein diet. Short-term supplementation of low (5%) protein diet (LPD) and high (40%) protein diet (HPD) produced no significant change in the HgluR binding and Bmax in young as well as aged rats. Consumption of LPD for long-term period (LTP) increased the HgluR binding by increasing Bmax in young rats; whereas, intake of the same diet by the aged rats under similar condition reduced the HgluR binding by reducing Bmax. Unlike LPD, HPD intake by the young rats for LTP decreased the HgluR binding by decreasing affinity (1/KD) without altering Bmax. However, the aged rats when supplemented with HPD for LTP, the HgluR binding was increased due to increase in Bmax. Further, no age-induced increase in HgluR binding and Bmax was observed following short-term supplementation of LPD; whereas, long-term intake of LPD decreased the HgluR binding and Bmax in 18 months old rats compared to 3 months old rats under similar condition. On the other hand, HPD under LTP potentiated the age-induced increase in HgluR binding and Bmax. These results, thus, suggest that dietary protein affects the hypothalamic glutamatergic activity by modulating its receptor population depending on the age of the subject, quantity of dietary protein and its duration of supplementation. PMID- 11478742 TI - Acetyl- and butyrylcholinesterase in normal and diabetic rat retina. AB - We studied the composition of molecular forms of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in normal and streptozotocin-induced diabetic rat retinas. Tissues were sequentially extracted with saline (S1) and saline detergent buffers (S2). 50% decrease in the amphiphilic G4 and G1 AChE molecular forms was observed in the diabetic retina compared to the controls. Less than 5% of the cholinesterase activity was due to BChE. 60% of the BChE activity in normal retina was brought into solution and evenly distributed between S1 and S2. In spite of the low BChE activity in the retina it was possible to detect globular forms (G(A)1, G(A)2, G(A)4, G(H)4) and a small proportion of an asymmetric form (A12) in the S1 extract. The G(A)4 and G(A)1 forms were found in the S2 extract. In the diabetic retina the activity of G(A)4 and G(A)1 BChE molecular forms was reduced 60% and 40% respectively. Our results indicate that diabetes caused a remarkable decrease in the activity of cholinesterase molecular forms in the retina. These decrease might participate in the alterations observed in the diabetic retina. PMID- 11478743 TI - Study of developmental changes on hexoses metabolism in rat cerebral cortex. AB - We have studied the developmental changes of glucose, mannose, fructose and galactose metabolism in rat cerebral cortex. As the animals aged, glucose, mannose and fructose oxidation to CO2 increased, whereas galactose oxidation decreased. Lipid synthesis from glucose and fructose also increased with age, that from mannose decreased and galactose did not change. Cytochalasin B, a potent non-competitive inhibitor of sodium-independent glucose transport, significantly impaired glucose, mannose and galactose metabolism, but had no effect on fructose metabolism. Both galactose or fructose did not change, whereas mannose declined the glucose metabolism. Glucose decreased fructose, galactose and mannose metabolism. Our results show that besides glucose, the metabolism of mannose, galactose and fructose present developmental changes from fetal to adult age, and reinforce the literature data indicating that mannose and galactose are transported by glucose carriers, while fructose is not. PMID- 11478744 TI - Effects of acetyl-L-carnitine on the formation of fatty acid ethyl esters in brain and peripheral organs after short-term ethanol administration in rat. AB - Increasing evidence suggests that Fatty acid ethyl esters (FAEE) play a central role in ethanol induced organ damage. In the current study we measured FAEE formation in rats after short-term oral administration of ethanol, in the presence and absence of pre-treatment with acetyl-L-carnitine. Ethanol treatment caused a significant increase in the levels of FAEE, particularly in the brain and heart, but also in the kidney and liver. Increases in FAEE were associated with a significant increase in FAEE synthase activity, GSH transferase activity, and lipid hydroperoxide levels. Pretreatment with acetyl-L-carnitine resulted in a significant reduction of FAEE accumulation, decrease in FAEE synthase and GSH transferase activities, and lipid hydroperoxide levels. Administration of acetyl L-carnitine greatly reduced the metabolic abnormalities due to non-oxidative ethanol metabolism, through an increment in lipid metabolism/turnover and by the modulation of the activities of enzymes associated with FAEE synthesis. These results suggest a potentially important pharmacological role for acetyl-L carnitine in the prevention of alcohol-induced cellular damage. PMID- 11478745 TI - Metabotropic glutamate receptors modulate GABA release from mouse hippocampal slices. AB - The effects of metabotropic glutamate receptor agonists on the basal and potassium (50 mM K+)-stimulated release of [3H]GABA from mouse hippocampal slices were investigated using a superfusion system. The group I agonist (1+/-)-1 aminocyclopentane-trans-1,3-dicarboxylate enhanced the basal GABA release and reduced the K+-evoked release by a mechanism antagonized by (RS)-1-aminoindan-1,5 dicarboxylate in both cases. The group II agonist (2S,2'R,3'R)-2-(2',3' dicarboxycyclopropyl)glycine failed to have any effect on the basal release, but inhibited the stimulated release. This inhibition was not affected by the antagonist (2S)-2-ethylglutamate. The group III agonists L(+)-amino-4 phosphonobutyrate and O-phospho-L-serine inhibited the basal GABA release, which effects were blocked by the antagonist (RS)-2-cyclopropyl-4 phosphonophenylglycine. Moreover, the suppression of the K+-evoked release by L(+)2-amino-4-phosphonobutyrate was apparently receptor-mediated, being blocked by (RS)-2-cyclopropyl-4-phosphonophenylglycine. The results show that activation of metabotropic glutamate receptors of group I is able to potentiate the basal release of GABA, whereas activation of groups I and III receptors reduce K+ stimulated release in mouse hippocampal slices. PMID- 11478746 TI - Brain metabolic effects of acute nicotine. AB - (-)Nicotine acetylcholine receptors are located on both nerve cell bodies and synaptic terminals, are permeable to calcium, and function perhaps predominantly by facilitating the release of neurotransmitters and neuropeptides. The behavioral rewards from (-)nicotine and perhaps addiction appear to be related to dopamine release. 31P NMR analysis reveals subcutaneously administered ( )nicotine produces acute alterations in brain membrane phospholipid and high energy phosphate metabolism of Fischer 344 rats. These metabolic responses to ( )nicotine could contribute to nicotine's behavioral effects. PMID- 11478747 TI - Serum ganglioside patterns in multiple sclerosis. AB - The relative distribution of gangliosides was determined in the serum of 37 patients with multiple sclerosis (MS) and of 30 healthy subjects. There was a significant increase of GM1 and GD1a, and a decrease of GM3 proportion in the serum of relapsing-remitting MS patients (RRMS) during their first MS attack. The RRMS patients in relapse with a long duration of the disease had a significant decrease of GM1 and an increase of GD1a portion in the serum. An increase of GD1a, one of the major brain neuron ganglioside fraction, suggested the neuron injury in the early and with a long duration RRMS. The finding of an increase of GM1, the main human myelin ganglioside, during the first MS attack in RRMS patients confirms previous evidence for the possible involvement of gangliosides in the early pathological course of demyelination in MS. PMID- 11478748 TI - The biological impact of Helicobacter pylori colonization. AB - Helicobacterpylori causes persistent inflammation in the human stomach, yet only a minority of persons harboring this organism develop peptic ulcer disease or gastric malignancy. H. pylori isolates possess substantial phenotypic and genotypic diversity, which may engender differential host inflammatory responses that influence clinical outcome. For example, strains that possess the cag pathogenicity island induce more severe gastritis and augment the risk for developing peptic ulcer disease and distal gastric cancer. However, important geographic differences in susceptibility to disease exist as clear-cut markers for H. pylori strains that affect certain groups of colonized individuals have little or no predictive power for other populations. Recent investigations that have more precisely delineated mechanisms of H. pylori pathogenesis will ultimately help to define which colonized persons bear the highest risk for subsequent development of clinical disease, and thus enable physicians to appropriately focus diagnostic testing and eradication therapy. Although stratification of disease risk based on H. pylori strain characteristics is unlikely to completely account for differences in clinical outcomes, it is an important first step in helping to understand the biology of long-term interactions between H. pylori and its human host. PMID- 11478749 TI - Treatment of Helicobacter pylori infection. AB - Helicobacter pylori is a serious, chronic, progressive, and transmissible infection associated with a significant morbidity and mortality, which alone emphasizes the priority of developing adequate prophylactic or therapeutic measures. What was previously termed "asymptomatic H. pylori infection" is now recognized as a latent infection, and it is now accepted that the presence of an H. pylori infection is an indication for eradication therapy. Successful cure of H. pylori infection requires 2 or more antibiotics. Antibiotic resistance is the major impediment of cure. The ideal duration of therapy is unknown, but in general, 1 week therapy is less effective than longer durations. Compliance is important for the success of treatment; therefore, the favored regimen should have the least side effects. At present, a proton pump inhibitor (or ranitidine bismuth citrate)-clarithromycin triple therapy with either amoxicillin or metronidazole, for at least 10 days is considered first-line therapy. The alternative is quadruple therapy containing a proton pump inhibitor, bismuth, tetracycline, and a higher dose of metronidazole. Quadruple therapy is the best choice after failure of proton pump inhibitor-clarithromycin triple therapy. Confirmation of successful therapy with a urea breath test or a stool antigen test is now the standard of practice. PMID- 11478750 TI - Is nonulcer dyspepsia related to Helicobacter pylori infection? AB - Dyspepsia, defined as pain or discomfort centered in the upper abdomen, is a common clinical problem. A variety of underlying disease states may result in dyspepsia, but commonly, diagnostic investigation will show no identifiable pathology, and the patient is diagnosed with nonulcer dyspepsia. Numerous hypothesis have been suggested as to the cause of symptoms in patients with nonulcer dyspepsia, including perturbations of gastroduodenal motility, hypersensitivity to physiologic stimuli including acid, and the effect(s) of infection within the gastric mucosa by Helicobacter pylori. Some epidemiological studies have suggested that patients with nonulcer dyspepsia may have a slightly higher prevalence of H. pylori infection. However, association does not prove causation. Causation of nonulcer dyspepsia by H. pylori could best be documented by resolution of symptoms following eradication of the infection. Early intervention studies indicated that there was a beneficial effect on symptoms of nonulcer dyspepsia with H. pylori eradication, but most of these studies had serious methodological flaws. In the last few years there have been a number of well-designed studies investigating the effect of H. pylori eradication on symptoms in patients with nonulcer dyspepsia. The results of these studies are inconsistent, but suggest that there is little, if any benefit from treatment. This case-based article on nonulcer dyspepsia discusses these studies in detail and provides a possible explanation for the differences in outcomes. PMID- 11478751 TI - The possible role of Helicobacter pylori in GERD. AB - A variety of abnormalities contribute to the development of gastroesophageal reflux disease (GERD) including transient lower esophageal sphincter relaxation, low esophageal sphincter pressure, presence of a hiatal hernia, diminished esophageal clearance of refluxed gastric contents, and alterations in esophageal mucosal resistance. Helicobacter pylori infection clearly plays a role in the pathogenesis of peptic ulcer disease and mucosa associated lymphoma of the stomach and is a definite risk factor for distal gastric cancer. The role of H. pylori infection in GERD remains controversial and incompletely understood. Although H. pylori infection does not cause reflux disease, circumstantial evidence suggests that it may protect against the development of GERD and its complications in some patients. The most likely mechanism whereby H. pylori infection protects against GERD is by decreasing the potency of the gastric refluxate in patients with corpus predominant gastritis. A variety of implications of H. pylori infection on GERD treatment have also arisen in recent years. These focus on the risk of gastric atrophy while on proton pump inhibitor therapy and the efficacy of proton pump inhibitors before and after eradication of H. pylori. This article puts into perspective our current understanding of the complex, incompletely understood relationship between H. pylori infection and GERD. PMID- 11478752 TI - Chronic gastritis and Helicobacter pylori. AB - Helicobacter pylori is the major cause of chronic gastritis. The predominant anatomic distribution of the gastritis is antral in the majority of individuals. In a small minority, the corpus is predominantly involved. The former pattern is associated with duodenal ulceration in some patients, but the majority of those infected never develop either symptoms or disease. The latter form is associated with the development of gastric ulcer and carcinoma and may be protective against the development of Barrett's esophagus. It is the physiological changes associated with the histological changes and the, as yet poorly, defined host response, which are of paramount importance in determining the evolution of a disease or whether the infected individual remains asymptomatic and disease free. This article addresses the various relationships between H. pylori infection, histology, gastric physiology, and disease. PMID- 11478753 TI - Helicobacter pylori and risk for gastric adenocarcinoma. AB - Gastric cancer is the second most common cause of cancer death in the world. Helicobacter pylori infection is now a well-accepted cause of this malignancy; in some parts of the world, up to eighty percent of all gastric cancers are at least in part caused by H. pylori infection. H. pylori infection typically starts in childhood as an inflammatory process in the stomach. The changes in the gastric microenvironment facilitate gastric cancer over time. Among infected individuals, genotype of H. pylori, coincident environmental exposures, and genetic factors of host seem to play roles in determining who will get gastric cancer and who will not. Unfortunately, it remains unknown whether treatment of H. pylori prevents gastric cancer. Thus, screening for H. pylori to prevent cancer is not yet widely recommended. Some consensus groups, however, have recommended screening for and treating H. pylori infection in individuals with family histories of gastric malignancy. In high-risk countries, screening programs for early gastric cancer itself may improve therapeutic outcome for this highly lethal disease. PMID- 11478754 TI - Reproductive and Child Health (RCH): an integrated approach--need of the day. PMID- 11478755 TI - Safe motherhood--a long way to achieve. AB - Maternal mortality ratio (MMR) is the most important index for monitoring the progress of safe motherhood programmes. A retrospective analysis of all maternal deaths at RG Kar Medical College and Hospital, Calcutta from 1st January, 1995 to 31st December, 1997 was carried out. There were 29,563 live births and 203 maternal deaths giving cumulative MMR of 686.67 per 100,000 live births. Among the victims 25.6% were < or = 20 years of age, majority (73.4%) were unbooked, mostly from rural (59.6%) or urban slum (20.2%) and from low socio-economic status (59.6%). Most (60.10%) were multiparous and 50.74% patients died within 24 hours of hospital admission. Direct causes were responsible for 83.25%, indirect causes for 14.78% and unrelated causes for 1.97% of maternal deaths. Toxaemia was the leading cause (53.20%) of maternal mortality. Other important causes were haemorrhage (16.75%), sepsis (12.31%), severe anaemia (6.4%), infective hepatitis (1.47%) and heart disease (3.44%). In comparison to previous years no significant changes in MMR had been found though there were temporary decline in some years. The contribution of toxaemia in maternal deaths is significantly high and is on the rise. The other national and global pictures were reviewed. India is among the countries of high MMR and far away from achieving safe motherhood. Majority of deaths are preventable. The medical causes of maternal deaths are in fact, the end point of a longer chain of many underlying factors like low socio-economic status, poverty, illiteracy, high parity and inadequate or no health care. An active community-based health care system accessible to every mother with strong referral system is the most effective approach to achieve safe motherhood. PMID- 11478756 TI - Male involvement in reproductive health care. AB - The programme of action globally endorsed at the International Conference on Population and Development (ICPD) emphasised the need for equity in gender relations with a special focus on men's shared responsibility and active involvement to promote reproductive and sexual health. If men are brought into a wide range of reproductive health services in such a way that they are supported as equal partners and responsible parents, as well as clients in their own right, better outcomes are expected in reproductive health indicators such as contraception acceptance and continuation, safer sexual behaviours, use of reproductive health services, and reduction in reproductive morbidity and mortality. This paper focuses on these key questions. What does men's involvement mean and how should it be operationalised? What does shared responsibility mean for various reproductive health problems subsumed within the reproductive health framework? Programmes to involve men should be designed to address three major goals: (1) Improve sexual and reproductive health of men and women, (2) generate men's support for women's actions related to reproduction and respect for women's reproductive and sexual rights, and (3) promote responsible and healthy reproductive and sexual behaviour in young men and boys. Gender inequality is a major barrier that must be overcome if these goals are to be met. Improving the reproductive well-being of women and men requires freeing them both from restricted gender roles. PMID- 11478757 TI - Reproductive health needs and care seeking behaviour of pavement dwellers of Calcutta. AB - An unabated growth of street dwellers in the city of Calcutta is reported to be due to twin reasons like, migration of rural poor people as well as uncontrolled fertility among these poor settlers of the city. A community-based study on reproductive health, fertility and related care seeking behaviour was studied among a sample of women of child bearing age living on streets of Calcutta. Besides, the quite common conditions like leucorrhoea (28.5%), menstrual irregularities (12.3%), infertility (2.5%) and STDs (1.3%) were also reported. But most of these illnesses (three-fourth) were uncared for, and the remaining one-fourth sought treatment from govemment institutions, private agencies or even from untrained practitioners (quacks). The reproductive behaviour of street dwelling women was characterised by early marriage, teenage pregnancies, and scarce use of contraceptives (32%) as well as frequent abortions (2.8%). Very few pregnant women received adequate antenatal care (3.8%). Coverage of tetanus toxoid immunisation (68.5%) and proper iron and folic acid supplementation (16.7%) were also poor. Whereas, antenatal care was received mostly from government health institutions (71%), home delivery (ie, on street) was a common practice and conducted mostly by untrained birth attendants (51.8%). PMID- 11478758 TI - Reproductive and Child Health (RCH) care and its implementation by IMA. AB - Reproductive and Child Health (RCH) is extended maternal child health of family welfare or safe motherhood or child survival and safe motherhood programme. Unless IMA members take up implementation of RCH programme, population stabilisation in India will remain unfulfilled. PMID- 11478759 TI - Maternal mortality in India. AB - Authors worked on maternal mortality in India from 1992-94. This article is a review article on the basis of authors' experience. The article starts with the brilliant sentence, 'Maternal mortality in India is unacceptably high' and ends with the suggested strategies to solve the problem. PMID- 11478760 TI - Endothelial dysfunction: therapy revisited (brief excerpts) (round table discussion on 18/01/2001 at Mumbai). AB - Indians have highest incidence of coronary heart disease. Here it occurs much more prematurely. Endothelial dysfunction has been increasingly recognised as an early event for the development of various cardiovascular disease. Long acting nifedipine has a definite role to play in reversing endothelial dysfunction. PMID- 11478761 TI - Evaluation of efficacy and safety of iron polymaltose complex and folic acid (Mumfer) vs iron formulation (ferrous fumarate) in female patients with anaemia. AB - Treatment of iron deficiency anaemia with conventional oral preparations is handicapped by unpredictable haematological response in addition to potential for irritating gastrointestinal adverse events. Iron polymaltose complex (IPC), a novel oral iron formulation with better absorbability, predictable haematinic response and less side effects was compared with oral ferrous fumarate in 100 female patients with documented iron deficiency anaemia. Clinical parameters (pallor, weakness) as well as biochemical parameters (Hb, serum iron, total iron binding capacity) show favourable changes with IPC; the physician and patient assessment also favour IPC over ferrous fumarate. PMID- 11478762 TI - Supportive role of a micronutrient appetite normaliser in convalescing patients. AB - A total of 722 patients recruited from various clinics across Western India, particularly in Western Maharashtra and South Gujarat, recovering from a variety of clinical conditions such as infections of the respiratory tract, malaria, enteric fever, jaundice, surgical procedures and so on were administered a micronutrient appetite normaliser formulation (neogadine) for periods up to 30 days. There was a significant and steady increase in the scores of appetite and well-being during the study period. Very good to good response were reported in 83% of patients. Neogadine was well tolerated with no significant adverse effects on any patients. Based on past experience with this micronutrient formulation as well as the findings of the present study, it is reasonable to conclude that neogadine plays a useful supportive role in the restoration of appetite and well being in conditions that apply to private practice situations in India. PMID- 11478763 TI - Intercellular mechanotransduction: cellular circuits that coordinate tissue responses to mechanical loading. AB - Physical forces play an important role in modulating cell function and shaping tissue structure. Mechanotransduction, the process by which cells transduce physical force-induced signals into biochemical responses, is critical for mediating adaptations to mechanical loading in connective tissues. While much is known about mechanotransduction in cells involving forces delivered through extracellular matrix proteins and integrins, there is limited understanding of how mechanical signals are propagated through the interconnected cellular networks found in tissues and organs. We propose that intercellular mechanotransduction is a critical component for achieving coordinated remodeling responses to force application in connective tissues. We examine here recent evidence on different pathways of intercellular mechanotransduction and suggest a general model for how multicellular structures respond to mechanical loading as an integrated unit. PMID- 11478764 TI - ERK pathway positively regulates the expression of Sprouty genes. AB - Sprouty was originally identified as an inhibitor of Drosophila development associated receptor tyrosine kinase (RTK) signaling. Although RTK signaling has been shown to induce Sprouty gene expression, the precise induction pathway downstream of RTK remains unclear. As RTK signaling pathway includes activation of extracellular signal-regulated kinases (ERKs), we have examined a correlation between activation of ERKs and induction of Sprouty gene expression. All reagents which induce the activation of ERKs induce Sprouty gene expression; these agents include not only growth factors which bind to RTK but also phorbol 12-myristate 13-acetate and active Raf-1 kinase. Furthermore, the Sprouty gene expression induced by all those agents is totally suppressed when the cells are pretreated with specific inhibitors of ERK kinase (MEK). Human tumor cells which exhibit constitutive activation of ERKs show elevated expression of Sprouty genes, which is abolished by treatment of these cells with MEK inhibitors. All these findings clearly indicate that Sprouty gene expression is positively regulated by the ERK pathway downstream of RTK. PMID- 11478765 TI - Human granulosa-lutein cells express functional EP1 and EP2 prostaglandin receptors. AB - Prostaglandin E(2) (PGE(2)) exerts mainly luteotrophic effects in the corpus luteum. In other tissues, PGE(2) acts via specific PGE(2) receptor subtypes including EP1, which modulates intracellular calcium ([Ca(2+)](i)) and EP2, which is coupled to cyclic AMP (cAMP) generation. We have therefore investigated the presence of functional EP1 and EP2 receptors using human granulosa-lutein (GL) cells. Reverse-transcription PCR revealed that GL cells expressed mRNA transcripts encoding both EP1 and EP2 receptors. When GL cells were challenged with ligands that can bind to both receptor subtypes (PGE(2) and 16,16 dimethyl PGE(2)) or exclusively to EP2 (butaprost), both cAMP formation and progesterone synthesis were stimulated. Furthermore, the cAMP response to these agonists could be significantly blocked by an EP1/2 antagonist AH6809 but not by an EP1 selective antagonist SC19220. Exposure of GL cells to 16,16-dm PGE(2) transiently raised [Ca(2+)](i) levels, which could be prevented by both AH6809 and SC19220. We therefore conclude that human GL cells express functional EP1 and EP2 receptors. PMID- 11478766 TI - Thrombopoietin induces histidine decarboxylase gene expression in c-mpl transfected UT7 cells. AB - The leukemic cell line UT7 is endowed with both megakaryocyte and basophil differentiation potential, as judged by its capacity to respond to PMA by displaying megakaryocytic and basophilic markers and to produce histamine by neosynthesis. Herein, we addressed the question whether the biological activities characteristic of basophil differentiation were still induced when c-mpl transfected UT7 cells received a specific megakaryocytic differentiation signal delivered by thrombopoietin (TPO). Surprisingly, we found that histamine synthesis did effectively occur in response to the growth factor. This activity was not associated with megakaryopoiesis since it was not detected in megakaryocytes generated from CD34(+) cells cultured in the presence of TPO. Comparing different c-mpl-transfected cell lines, we found that the amount of histamine generated in response to TPO correlated with their responsiveness to PMA, but not with their level of c-mpl expression, thus revealing an intrinsic basophil differentiation potential. Both PMA- and TPO-induced histamine synthesis was reduced by PKC and MEKs inhibitors, indicating that the induction occurred through a common signalling pathway. PMID- 11478767 TI - Apoptosis induction by epigallocatechin gallate involves its binding to Fas. AB - Epigallocatechin gallate (EGCG) is known to induce apoptosis in various types of tumor cells, but the precise mechanism by which EGCG induces apoptosis remains to be elucidated. The Fas-Fas ligand system is one of the major pathways operating in the apoptotic cascade. The aim of this study was to examine the possibility that EGCG-binding to Fas triggers the Fas-mediated apoptosis. The EGCG treatment of human monocytic leukemia U937 cells resulted in elevation of caspase 8 activity and fragmentation of caspase 8. The DNA ladder formation caused by the EGCG treatment was inhibited by the caspase 8 inhibitor. These findings suggested the involvement of the Fas-mediated cascade in the EGCG-induced apoptosis in U937 cells. Affinity chromatography revealed the binding between EGCG and Fas. Thus, the results suggest that EGCG-binding to Fas, presumably on the cell surface, triggers the Fas-mediated apoptosis in U937 cells. PMID- 11478768 TI - Kynurenine binds to the peptide binding region of the chaperone alphaB crystallin. AB - UV filters, such as kynurenine, are present in the human lens. They are spontaneously unstable at neutral pH and deaminate to form reactive alpha, beta unsaturated ketones. This process becomes more prominent after the lens barrier develops in middle age. Here we show that deaminated kynurenine reacts primarily with histidine residues in alphaB-crystallin: a major lens protein that lacks cysteine. Five of the nine histidines in alphaB-crystallin were found to be conjugated with kynurenine. Furthermore, a major site of covalent modification was at histidine 83, which is found in the putative peptide binding region of alphaB-crystallin; a site crucial for its role as a chaperone. We propose that modification of alphaB-crystallin by UV filters may compromise the chaperone action of this protein. PMID- 11478769 TI - Effects of capsaicin on Ca(2+) release from the intracellular Ca(2+) stores in the dorsal root ganglion cells of adult rats. AB - We have investigated the effect of capsaicin on Ca(2+) release from the intracellular calcium stores. Intracellular calcium concentration ([Ca(2+)](i)) was measured in rat dorsal root ganglion (DRG) neurons using microfluorimetry with fura-2 indicator. Brief application of capsaicin (1 microM) elevated [Ca(2+)](i) in Ca(2+)-free solution. Capsaicin-induced [Ca(2+)](i) transient in Ca(2+)-free solution was evoked in a dose-dependent manner. Resiniferatoxin, an analogue of capsaicin, also raised [Ca(2+)](i) in Ca(2+)-free solution. Capsazepine, an antagonist of capsaicin receptor, completely blocked the capsaicin-induced [Ca(2+)](i) transient. Caffeine completely abolished capsaicin induced [Ca(2+)](i) transient. Dantrolene sodium and ruthenium red, antagonists of the ryanodine receptor, blocked the effect of capsaicin on [Ca(2+)](i). However, capsaicin-induced [Ca(2+)](i) transient was not affected by 2-APB, a membrane-permeable IP(3) receptor antagonist. Furthermore, depletion of IP(3) sensitive Ca(2+) stores by bradykinin and phospholipase C inhibitors, neomycin, and U-73122, did not block capsaicin-induced [Ca(2+)](i) transient. In conclusion, capsaicin increases [Ca(2+)](i) through Ca(2+) release from ryanodine sensitive Ca(2+) stores, but not from IP(3)-sensitive Ca(2+) stores in addition to Ca(2+) entry through capsaicin-activated nonselective cation channel in rat DRG neurons. PMID- 11478770 TI - The AH receptor of the most dioxin-sensitive species, guinea pig, is highly homologous to the human AH receptor. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) brings about a wide spectrum of toxic and biochemical changes, most of which are mediated by the AH receptor (AHR). Recent cloning of the AHR from the two most TCDD-resistant laboratory animals, Han/Wistar (Kuopio) rats and hamsters, suggested a critical role for the C terminal transactivation domain structure in TCDD sensitivity. Here we cloned the AHR from the most TCDD-susceptible species, guinea pig. The N-terminus of its AHR was highly similar to that in the resistant animals. However, the C-terminal Q rich subdomain was only about half the size of this subunit in the hamster AHR. There was a distinct correlation across published mammalian species between the number of glutamine residues in the Q-rich subdomain and sensitivity to the acute lethality of TCDD. The closest homolog of the Guinea pig receptor turned out to be the human AHR, which may be relevant for dioxin risk assessment. PMID- 11478771 TI - Upregulation of vascular NAD(P)H oxidase subunit gp91phox and impairment of the nitric oxide signal transduction pathway in hypertension. AB - In this study we analyzed the role of vascular NAD(P)H oxidase in the generation of O(2)(-) and the endothelial impairment of NO signal transduction pathway in hypertension. In aortic rings of 15-month-old stroke-prone spontaneously hypertensive rats (SHR15) we found a 10-fold increased expression of NAD(P)H oxidase subunit gp91phox mRNA associated with a 3-fold increased production of O(2)(-) compared to age-matched Wistar rats (WIS15). Vasorelaxation studies in aortas of SHR15 showed a strongly diminished response to acetylcholine, NO-donor S-nitroso-N-acetyl-d,l-penicillamine, and organic nitrate glyceryl trinitrate compared to WIS15. Soluble guanylate cyclase (sGC) activity and sGC beta(1) subunit protein expression was downregulated in aortas and lungs of SHR15. These data suggest an upregulation of vascular NAD(P)H oxidase and an impairment of the NO signal transduction pathway in hypertension. PMID- 11478772 TI - Trifluoroacetic acid activates ATP-sensitive K(+) channels in rabbit ventricular myocytes. AB - Recent in vivo experimental evidence suggests that isoflurane-induced cardioprotection may involve K(ATP) channel activation during myocardial ischemia. The actual effect of isoflurane on cardioprotective ion conductance, however, such as that mediated by the opening of K(ATP) channels, has been the subject of some controversy in the past. The investigation reported here used a patch-clamp technique to test the hypothesis that a metabolite of isoflurane, trifluoroacetic acid (TFA), contributes to isoflurane-induced cardioprotection via K(ATP) channel activation. TFA enhanced channel activity in a concentration dependent fashion, exhibiting half-maximal activation at 0.03 mM. TFA increased the number of openings of the channel, but did not affect the single channel conductance of K(ATP) channels. Analysis of open and closed time distributions showed that TFA increased the burst duration and decreased the interburst interval without eliciting changes of less than 5 ms in open and closed time distributions. TFA diminished the ATP sensitivity of K(ATP) channels in a concentration-response relationship for ATP. These results imply that TFA could mediate isoflurane-induced cardioprotection via K(ATP) channel activation during myocardial ischemia and reperfusion. PMID- 11478773 TI - Dna fragmentation factor 45 mutant mice exhibit resistance to kainic acid-induced neuronal cell death. AB - Excitotoxicity is a process where glutamate or other excitatory amino acids induce neuronal cell death. Emerging evidence suggests that apoptosis plays a key part in excitotoxic neurodegeneration. The DNA fragmentation factor 45 (DFF45 or ICAD) is a subunit of a heterodimeric DNase complex crucial for DNA fragmentation during apoptosis. Using a DFF45 mutant mouse model, we previously found that DFF45 deficient cells are more resistant to apoptosis than normal control cells. To investigate whether the lack of DFF45 may attenuate neuronal cell death induced by excitotoxicity, we compared kainic acid-induced seizure behavior and neuronal cell death in DFF45 mutant and wild-type control mice. We found that the mutant mice exhibit similar kainic acid-induced seizure severity compared to control mice. However, DFF45 mutant mice are more resistant than control mice to kainic acid-induced CA3 neuronal cell death. Interestingly, residual DNA degradation can be detected in the hippocampus of DFF45 mutant mice that exhibit KA-induced lesions. Our results suggest that a lack of DFF45 can lead to neuronal resistance to excessive activity-induced toxicity. PMID- 11478774 TI - Evidence that caspase-13 is not a human but a bovine gene. AB - Caspase-13 was reported to be a member of the human caspase family of proteases (Humke, E., et al., J. Biol. Chem. 273, 15702-15707, 1998). By contrast, a recent study (Lin, X., et al., J. Biol. Chem. 275, 39920-39926, 2000) could not confirm caspase-13 expression in human tissues. When we searched the GenBank database we found several expressed sequence tags (ESTs) from bos taurus completely matching the published caspase-13 sequence. Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that bovine but not human peripheral blood mononuclear cells express caspase-13. From these cells we cloned two bovine caspase-13 splice variants and found that the sequence of the larger variant was identical to the mRNA published by Humke et al. Our findings strongly suggest that the previously published caspase-13 sequence is not of human origin but represents a bovine gene. PMID- 11478775 TI - Pivotal role of mitochondrial Ca(2+) in microcystin-induced mitochondrial permeability transition in rat hepatocytes. AB - We have shown earlier that microcystin-LR (MLR), a specific hepatotoxin, induced onset of mitochondrial permeability transition (MPT) and apoptosis in rat hepatocytes. Here we attempted to investigate the role of mitochondrial Ca(2+) in MLR-induced onset of MPT and cell death. Using confocal microscopy, we found that MLR caused an early surge of mitochondrial Ca(2+) prior to the onset of MPT and cell death. Pretreatment with 1,2-bis(O-aminophenoxyl)ethane-N,N,N',N'-tetracetic acid tetra(acetoxymethyl)ester (an intracellular Ca(2+) chelator) or ruthenium red (an inhibitor of mitochondrial Ca(2+) uniporter) prevented the early mitochondrial Ca(2+) surge and attenuated the subsequent onset of MPT and cell death. On the other hand, a mitochondrial uncoupler, CCCP, rapidly disrupted the mitochondrial membrane potential and also prevented the mitochondrial Ca(2+) surge, onset of MPT, and cell death. We thus conclude that mitochondrial Ca(2+) plays an important role in the onset of MPT and cell death in MLR-treated rat hepatocytes. PMID- 11478776 TI - Acivicin induces apoptosis independently of gamma-glutamyltranspeptidase activity. AB - Inhibition of cellular gamma-glutamyltranspeptidase (GGT) enzyme activity by its specific inhibitor acivicin is frequently used in studies aimed at demonstrating the physiological role of this enzyme. However, because acivicin is a glutamine antagonist, it also inhibits many other glutamine-dependent enzymes involved in purine and pyrimidine biosynthesis. The objective of the present work is to determine whether acivicin exhibits apoptotic properties and the significance of GGT activity level in the response to acivicin treatment. We compared acivicin (0 150 microM) effect on V79 cell lines expressing or not expressing human GGT. Apoptosis was assayed by annexin-V staining, cell cycle analysis, and caspase activation using flow cytometry. We found that acivicin causes a dose- and time dependent apoptosis in the GGT-negative V79 cell line as well as in its GGT positive counterpart line. This is the evidence that acivicin induces apoptosis in V79 cell independently of their GGT activity level. PMID- 11478777 TI - Dynamic compression inhibits the synthesis of nitric oxide and PGE(2) by IL-1beta stimulated chondrocytes cultured in agarose constructs. AB - Both mechanical loading and interleukin-1beta (IL-1beta) are known to regulate metabolic processes in articular cartilage through pathways mediated by nitric oxide ((*)NO) and PGE(2). This study uses a well-characterized model system involving isolated chondrocytes cultured in agarose constructs to test the hypothesis that dynamic compression alters the synthesis of (*)NO and PGE(2) by IL-1beta-stimulated articular chondrocytes. The data presented demonstrate for the first time that dynamic compression counteracts the effects of IL-1beta on articular chondrocytes by suppressing both (*)NO and PGE(2) synthesis. Inhibitor experiments indicated that the dynamic compression-induced inhibition of PGE(2) synthesis and stimulation of proteoglycan synthesis were (*)NO mediated, while compression-induced stimulation of cell proliferation was (*)NO independent. The inhibition of (*)NO and PGE(2) by dynamic compression is a finding of major significance that could contribute to the development of novel strategies for the treatment of cartilage-degenerative disorders. PMID- 11478778 TI - Sulfation of flavonoids and other phenolic dietary compounds by the human cytosolic sulfotransferases. AB - The protective effects of diet, especially soya products, tea, and many fruits, against a variety of human cancers, as suggested by epidemiological studies, has focused attention on flavonoids, isoflavonoids, and other phenolic dietary compounds as chemoprotectants. Among the mechanisms suggested for their chemoprotective action, their ability to inhibit the bioactivation of carcinogens by the human cytosolic sulfotransferases (STs) and the direct effects of their sulfoconjugates are being increasingly studied. We report here a systematic study on the sulfation of representative flavonoids, isoflavonoids, anti-oxidants, and other phenolic dietary compounds by all ten known human cytosolic STs. All ten recombinant human cytosolic STs were prepared in a pure form and tested for their sulfating activities with a variety of these compounds. P-form (SULT1A1) phenol ST (PST) showed high sulfating activity with most of these compounds. M-form (SULT1A3) PST showed high activity with the flavonoids but not with the isoflavonoids. SULT1C ST #2 showed high activity with the isoflavonoids and also sulfated most of the other compounds. Possible relevance of these results to the chemoprotective effects of these dietary compounds is discussed. PMID- 11478779 TI - Ligand affinity, homodimerization, and ligand-induced secondary structural change of the human vitamin d receptor. AB - The intrinsic tryptophan fluorescence signal of the full-length nuclear receptor hVDR was used to directly determine the dissociation constants, K(d), of two ligands yielding K(d) = 32 nM for 1alpha,25(OH)(2)D(3) and K(d) = 322 nM for 25(OH)D(3). Ligand binding was accompanied by a conformational change in the alpha-helical part of hVDR as revealed by CD spectroscopy. In addition, the presence of calcitriol was found to be a necessary prerequisite for the homodimerisation of hVDR which was monitored using fluorescence anisotropy. We conclude that the observed ligand-induced structural change of hVDR is conditional for dimerisation of the protein. PMID- 11478780 TI - Ntg2 of Saccharomyces cerevisiae repairs the oxidation products of 8 hydroxyguanine. AB - In Escherichia coli, endonuclease III (endo III) repairs the oxidation products of 8-OHGua. However, the corresponding repair enzymes in eukaryotes have not been identified. Here we report that 8-hydroxyguanine (8-OHGua) is highly sensitive to further oxidation. We also show that Ntg2, a functional homolog of endo III in Saccharomyces cerevisiae, is capable of nicking the irradiated duplex DNA containing 8-OHGua. Moreover, Ntg2 formed a stable complex with the DNA upon incubation with NaBH(4). In contrast, Ntg1, another functional homolog of endo III, showed no such activities. These findings indicate that Ntg2 is, at least in part, responsible for repairing the oxidation products of 8-OHGua in eukaryotic cells. PMID- 11478781 TI - Caspase-mediated calcineurin activation contributes to IL-2 release during T cell activation. AB - Calcineurin, a Ca(2+)/calmodulin-dependent Ser/Thr phosphatase (protein phosphatase 2B), plays a critical role in IL-2 production during T cell activation. It has been previously reported that IL-2 release in activated Jurkat T requires caspase-like activity (Posmantur et al. (1998) Exp. Cell. Res. 244, 302-309). We report here that the 60-kDa catalytic subunit of calcineurin A (Cn A) was partially cleaved to a 45-kDa form in phytohemagglutinin A (PHA) or phorbol ester + ionomycin (P + I)-activated Jurkat cells. In parallel, proteolytic activation of upstream caspases (caspase-8 and -9) as well as effector caspase-3 was also observed. Cn A cleavage was caspase mediated, since it was inhibitable by pan-caspase inhibitor Cbz-Asp-CH(2)OC(O)-2,6 dichlorobenzene (Z-D-DCB). Cn A cleavage was also observed when purified calcineurin was digested in vitro with caspase-3. Truncated Cn A was associated with enhanced phosphatase activity and reduced calmodulin sensitivity. Furthermore, in PHA or P + I-activated Jurkat cells, dephosphorylation of calcineurin substrate NFATc (a transcription factor known to be involved in transactivation of the IL-2 gene), was also suppressed by Z-D-DCB. Taken together, our results suggest that caspase-mediated cleavage of Cn A contributes to IL-2 production during T cell activation. PMID- 11478782 TI - The transcription factor onecut-2 controls the microphthalmia-associated transcription factor gene. AB - Microphthalmia-associated transcription factor (MITF) is essential for melanocyte differentiation. MITF mutations are associated with some cases of Waardenburg syndrome (WS) type 2. WS is a dominantly inherited disease characterized by auditory-pigmentary defects that result from the absence of melanocytes. The lack of mutation in MITF coding sequences in some WS2 patients suggests that unidentified factors controlling MITF expression might be involved. We show here that the cut-homeodomain transcription factor Onecut-2 (OC-2) is expressed in melanocytes and binds to the MITF gene promoter. Overexpression of OC-2 in transfected cells stimulates MITF promoter activity. Mutations that prevent OC-2 binding decrease MITF promoter activity by 75%. Based on these results, we searched in 56 WS2 patients for mutations in the OC2 gene or in OC-2 binding sites in the MITF promoter, but none was found. These results show that OC-2 stimulates MITF expression and that OC2 is a candidate gene, but not a common cause, of WS. PMID- 11478783 TI - Lupus autoantibodies recognize the product of an alternative open reading frame of SmB/B'. AB - An unusual feature of the gene for the spliceosomal protein SmB/B' is the presence of an unusually long alternative open reading frame (aORF) which could encode 220 amino acids. We cloned and expressed this aORF protein and used immunological assays to determine its antigenicity in patients with systemic lupus. Sera from 10 of 22 (46%) anti-Sm positive lupus patients showed significant binding to the SmB' aORF protein by ELISA while neither the normal controls nor anti-Sm negative lupus patient controls showed significant reactivity. Antigenicity of the SmB' aORF protein was further localized to the C terminus using a deletion construct. This is the first known example in which the product of an alternative open reading frame acts as an autoantigen in human disease. These results are consistent with the possibility that generation of anti-Sm autoantibodies in a subset of lupus patients is due to abnormal processing and expression of an aORF SmB/B' message, by an as yet unidentified mechanism. PMID- 11478784 TI - The chloroplast-derived trnW and trnM-e genes are not expressed in Arabidopsis mitochondria. AB - Depending on their genetic origin, plant mitochondrial tRNAs are classified into three categories: the "native" and "chloroplast-like" mitochondrial-encoded tRNAs and the imported nuclear-encoded tRNAs. The number and identity of tRNAs in each category change from one plant specie to another. As some plant mitochondrial trn genes were found to be not expressed, and as all Arabidopsis thaliana mitochondrial trn genes are known, we systematically tested the expression of A. thaliana mitochondrial trn genes. Both the "chloroplast-like" trnW and trnM-e genes were found to be not expressed. These exceptions are remarkable since trnW and trnM-e are expressed in the mitochondria of other land plants. Whereas we could not conclude which tRNA(Met) compensates the lack of expression of trnM-e, we showed that the cytosolic tRNA(Trp) is present in A. thaliana mitochondria, thus compensating the absence of expression of the mitochondrial-encoded trnW. PMID- 11478785 TI - Preconditioning decreases ischemia/reperfusion-induced peroxynitrite formation. AB - The role for peroxynitrite (ONOO(-)) in the mechanism of preconditioning is not known. Therefore, we studied effects of preconditioning and subsequent ischemia/reperfusion on myocardial ONOO(-) formation in isolated rat hearts. Hearts were subjected to a preconditioning protocol (three intermittent periods of global ischemia/reperfusion of 5 min duration each) followed by a test ischemia/reperfusion (30 min global ischemia and 15 min reperfusion). When compared to nonpreconditioned controls, preceding preconditioning improved postischemic cardiac performance and significantly decreased test ischemia/reperfusion-induced formation of free nitrotyrosine measured in the perfusate as a marker for cardiac endogenous ONOO(-) formation. During preconditioning, however, the first period of ischemia/reperfusion increased nitrotyrosine formation, which was attenuated after the third period of ischemia/reperfusion. We conclude that classic preconditioning inhibits ischemia/reperfusion-induced cardiac formation of ONOO(-) and that subsequent periods of ischemia/reperfusion result in a gradual attenuation of ischemia/reperfusion-induced ONOO(-) generation. This mechanism might be involved in ischemic adaptation of the heart. PMID- 11478786 TI - Lecithinized Cu, Zn-superoxide dismutase limits the infarct size following ischemia-reperfusion injury in rat hearts in vivo. AB - Covalent binding of 4 molecules of phosphatidylcholine palmitoyl to human recombinant superoxide dismutase (SOD) results in a compound (lecithinized SOD) that has a longer half-life and greater affinity to the cell membrane than unmodified SOD. We investigated whether lecithinized SOD played a protective role against myocardial ischemia-reperfusion injuries in rats. Rats underwent 45 min of myocardial ischemia by occluding the left coronary artery followed by 120 min of reperfusion. They were randomly assigned to receive either lecithinized SOD, polyethylene glycol conjugated SOD (PEG-SOD), unmodified SOD, free lecithin derivative, or PBS intravenously at 5 min prior to reperfusion. Myocardial infarct area assessed by TTC staining was smaller in lecithinized SOD group than PEG-SOD, unmodified SOD, free lecithin derivative or control group. Blood pressure and heart rate was similar in each group. ELISA demonstrated SOD level in the heart was significantly high in lecithinized SOD group, especially in the heart of ischemia at risk. Although serum SOD level of PEG-SOD was as high as lecithinized SOD, SOD level of the heart was low. These data suggested lecithinized SOD had a protective effect in myocardial ischemia-reperfusion injuries through its increased bioavailability. PMID- 11478787 TI - Extracellular NAD(+) induces calcium signaling and apoptosis in human osteoblastic cells. AB - ADP-ribosyl cyclase/CD38 is a bifunctional enzyme that catalyzes at its ectocellular domain the synthesis from NAD(+) (cyclase) and the hydrolysis (hydrolase) of the calcium-mobilizing second messenger cyclic ADP ribose (cADPR). Furthermore, CD38 mediates cADPR influx inside a number of cells, thereby inducing Ca(2+) mobilization. Intracellularly, cADPR releases Ca(2+) from ryanodine-sensitive pools, thus activating several Ca(2+)-dependent functions. Among these, the inhibition of osteoclastic-mediated bone resorption has been demonstrated. We found that HOBIT human osteoblastic cells display ADP-ribosyl cyclase activity and we examined the effects of CD38 stimulation on osteoblasts function. Extracellular NAD(+) induced elevation of cytosolic calcium due to both Ca(2+) influx from the extracellular medium and Ca(2+) release from ryanodine sensitive intracellular stores. Culturing these cells in the presence of NAD(+) caused a complete growth arrest with a time-dependent decrease of cell number and the appearance of apoptotic nuclei. The first changes could be observed after 24 h of treatment and became fully evident after 72-96 h. We propose a role of extracellular NAD(+) in bone homeostatic control. PMID- 11478788 TI - Identification of N(omega)-carboxymethylarginine, a new advanced glycation endproduct in serum proteins of diabetic patients: possibility of a new marker of aging and diabetes. AB - A new advanced glycation end product (AGE), N(omega)-carboxymethyl-arginine (CMA), was found in acid-soluble skin collagen of a newborn bovine prepared by in vitro glycation with 1 M glucose incubation at 37 degrees C for about 30 days [ 1 ]. CMA production was increased with incubation time in parallel, and after 30 days incubation the yield was 100 times higher than that of pentosidine [ 1 ]. This result suggested the importance of CMA as a major AGE in collagen. We have detected and measured the CMA level in human serum proteins by electrospray ionization/liquid chromatography/mass spectrometry (ESI/LC/MS), using CMA standard concentration curve. In this report, we first show the existence of CMA in vivo, and its serum level is significantly elevated in diabetic serum proteins, compared to age-matched control serum proteins. These results provide strong evidence that CMA is a new diagnostic marker of glycation in diabetes. PMID- 11478789 TI - K252a, an indrocarbazole derivative, causes the membrane of myoblasts to enter a fusion-capable state. AB - K252a, an indrocarbazole derivative and protein kinase inhibitor, is reported to promote myogenic differentiation in C2 mouse myoblasts. We examined the effects of K252a on QM-RSV cells, quail myoblasts transformed with a temperature sensitive mutant of Rous sarcoma virus. K252a promoted myotube formation of QM RSV cells. Presumptive QM-RSV cells also formed multinucleated cells when exposed to K252a. However, the expression of myogenin, a muscle regulatory factor, was not stimulated in the presence of the drug, suggesting that it promotes membrane fusion but not myogenic differentiation. To confirm the promotion of membrane fusion by K252a, presumptive C2 cells, which are strongly resistant to HVJ mediated cell fusion, were fused by HVJ (Sendai virus) after K252a treatment. Presumptive C2 cells treated with K252a fused with HVJ, demonstrating that K252a causes the cells to enter a fusion-capable state. The amount of membrane cholesterol, a factor that decreases membrane fluidity, fell in K252a-treated C2 cells. The results suggest that a decrease of membrane cholesterol is a cause of the change that renders myoblast membrane susceptible to fusion in the presence of K252a. PMID- 11478790 TI - Construction of preferential cDNA microarray specialized for human colorectal carcinoma: molecular sketch of colorectal cancer. AB - cDNA microarray analyses can be used to identify candidate genes that play important roles in human carcinogenesis. To gain insight into the molecular sketch of colorectal cancer, we have constructed cDNA microarrays specialized for colorectal cancer, which we named "Colonochip" by selecting genes that are expressed in colorectal cancer, normal colonic mucosa, and liver metastatic cancer tissues. This microarray contained 4608 nonredundant cDNA clones from over 30,000 cDNA clones derived from the three types of human cDNA libraries, as well as clones from 170 additional conventional major genes suspected to be involved in colorectal carcinogenesis, according to literatures. Using this "Colonochip," we were able to identify 59 genes showing twofold or more differential expression between primary cancer and normal colonic mucosa, potent candidates for diagnosis, and therapy of colorectal cancer for further studies. PMID- 11478791 TI - Immunoreactivity of VR1 on epidermal keratinocyte of human skin. AB - We demonstrated the immunoreactivity of the receptor proteins, VR1, ion channels associated with pain sensation, on the epidermis of the human skin. Immunohistochemistry using antiserum against VR1 derived peptide showed immunoreactivity on the keratinocytes cell membrane of the human epidermis and cultured keratinocytes. The blocking peptide of the antiserum reduced the immunoreactivity on the epidermis. RT-PCR assay of cultured human keratinocyte also showed expression of VR1 mRNA. These results suggest the existence of VR1 like protein in epidermal keratinocytes of human skin. PMID- 11478792 TI - Polarized sorting of aquaporins 5 and 8 in stable MDCK-II transfectants. AB - Localization of aquaporin (AQP) water channels to either apical or basolateral membranes is important for various epithelial functions. We have established MDCK II cell transfectants stably expressing AQP5 (RW5 cells) or AQP8 (RW8 cells). The expression of both AQPs was confirmed by the results of immunofluorescence microscopy and immunoblot analysis. When grown on polycarbonate filters, osmotically-obliged transepithelial water flow across RW5 and RW8 monolayers was approximately 3-fold greater than that occurring across a monolayer of the parental cell line. Importantly, results of confocal immunofluorescence microscopy studies showed that AQP5 sorted to the apical membranes of RW5 cells. In contrast, AQP8 sorted to the basolateral membranes of RW8 cells. This is the first report of (i) stable epithelial cell cultures exhibiting a functional, polarized distribution of AQPs 5 and 8, and (ii) a basolateral localization of AQP8 in a polarized epithelial cell. PMID- 11478793 TI - 2-Methoxyestradiol induces G2/M arrest and apoptosis in prostate cancer. AB - Few therapeutic treatment options are available for patients suffering from metastatic androgen-independent prostate cancer. We investigated the ability of the estrogen metabolite 2-methoxyestradiol to inhibit the proliferation of a variety of human prostate cancer cell lines in vitro and to inhibit the growth of androgen-independent prostate cancer in a transgenic mouse model in vivo. Our results showed that 2-methoxyestradiol is a powerful growth inhibitor of LNCaP, DU 145, PC-3, and ALVA-31 prostate cancer cells. Cell flow cytometry of 2 methoxyestradiol-treated DU 145 cells showed a marked accumulation of cells in the G2/M phase of the cell cycle and an increase in the sub-G1 fraction (apoptotic). In addition, staining for annexin V, changes in nuclear morphology, and inhibition of caspase activity support a role for apoptosis. More importantly, we showed that 2-methoxyestradiol inhibits prostate tumor progression in the Ggamma/T-15 transgenic mouse model of androgen-independent prostate cancer without toxic side effects. These results in cell culture and an animal model support investigations into the clinical use of 2-methoxyestradiol in patients with androgen-independent prostate cancer. PMID- 11478794 TI - Differential binding of arfaptin 2/POR1 to ADP-ribosylation factors and Rac1. AB - We first identified arfaptin as a protein that bound to GTP-ARFs (especially ARF1). However, a second group reported that POR1, a truncated form of arfaptin, bound to GTP-Rac1. Therefore, we examined the possibility that arfaptin 2/POR1 was a common downstream effector for both ARF1 and Rac1. In this study, we found that constitutively active Rac1 or GTP-Rac1 showed negligible or no binding to arfaptin 2/POR1 in a yeast two-hybrid assay or a GST pull-down assay. However, wild-type or dominant negative Rac1 or Rac1 liganded to GDP showed strong binding. In contrast, constitutively active ARFs1, 5, and 6 showed binding, whereas the wild-type and dominant negative forms did not. Furthermore, the GTP liganded ARFs bound arfaptin 2, whereas the GDP-bound forms showed little or no binding. Based on these observations, we suggest that arfaptin 2/POR1 is a target protein for GTP-ARFs and for GDP-Rac1, and that it may be involved in interactions between the Rac and ARF signaling pathways. PMID- 11478795 TI - Sequence analysis identifies TTRAP, a protein that associates with CD40 and TNF receptor-associated factors, as a member of a superfamily of divalent cation dependent phosphodiesterases. AB - CD40 is a member of the tumor necrosis factor (TNF) receptor family. CD40 mediated signal transduction involves the recruitment of several cytoplasmic proteins and induces expression of a large number of genes. TTRAP, a novel protein that interacts with the cytoplasmic domain of CD40 and with TNF-receptor associated factors (TRAFs), has been cloned and shown to inhibit nuclear factor kappaB activation (NF-kappaB). By using various bioinformatics-based sequence and structure analyses of proteins involved in signaling by the TNF receptor family, we found that TTRAP is a member of a superfamily of Mg(2+)/Mn(2+)-dependent phosphodiesterases. More precisely, our results suggest that TTRAP is related to the human APE1, a Mg(2+)-dependent endonuclease. This potential novel function of TTRAP raises the intriguing possibility for a role of APE1-like DNA-repair endonucleases in TNF receptor family-mediated signaling and functions. PMID- 11478796 TI - Transcriptional response of a human colon adenocarcinoma cell line to sodium butyrate. AB - Taking advantage of the DNA array screening technology, we analysed the effect of sodium butyrate on mRNA transcription in human HT29 colon adenocarcinoma cells. Out of 588 mRNA species analysed, only 119 resulted expressed. Among these, 60 exhibited a variable degree of modulation after butyrate treatment. Genes linked to the cell growth, apoptosis and oxidative metabolism appeared the most significantly affected. Furthermore, many of the differentially expressed genes are transcription factors and this may account for the variability of the biological effects of butyrate. The pattern of butyrate-affected genes may represent a reference in further analyses of gene expression of intestinal cells and tissues. PMID- 11478797 TI - Recombinant hTASK1 is an O(2)-sensitive K(+) channel. AB - Hypoxic inhibition of background K(+) channels is crucial to O(2) sensing by chemoreceptor tissues, but direct demonstration of O(2) sensitivity by any member of this K(+) channel family is lacking. HEK293 cells were transfected with a pcDNA3.1-hTASK1 construct; expression of hTASK1 was verified using RT-PCR and immunocytochemistry. Whole-cell K(+) currents of cells stably expressing hTASK-1 were, as anticipated, extremely sensitive to extracellular pH, within the physiological range (IC(50) approximately 7.0). All cells expressing this signature pH sensitivity were acutely modulated by pO(2); reduction of pO(2) from 150 to <40 mmHg (at pH 7.4) caused rapid and reversible suppression of pH sensitive K(+) currents. Furthermore, these two regulatory signals clearly acted at the same channel, since the magnitude of the O(2)-sensitive current was dependent on the extracellular pH. These data represent the first direct verification that hTASK1 is O(2)-sensitive and reinforce the idea that this K(+) channel is key to O(2) sensing in chemoreceptors. PMID- 11478798 TI - ABCA6, a novel a subclass ABC transporter. AB - Here we report the cDNA cloning of a novel member of the ABC A transporter subfamily from human macrophages. The identified coding sequence is of 5.0 kb size and contains an open reading frame which encodes a 1617 amino acid polypeptide. Structurally, the putative ABC transporter protein product consists of two tandemly oriented subunits, each composed of a transmembrane domain followed by a nucleotide binding fold, and thus conforms to the group of full size ABC transporters. We also demonstrate the existence of an alternative transcript that codes for a 637 amino acid protein variant bearing the features of a truncated half-size transporter. Among the human ABC transporter subfamily A the novel transporter shows highest protein sequence homology with ABCA8 (60%), followed by ABCA2 (32%) and ABCA1 (32%), respectively. In agreement with the proposed classification, the novel transporter was designated ABCA6. The ABCA6 gene is ubiquitously expressed with highest mRNA levels in liver, lung, heart and brain. Analysis of the genomic organization demonstrated that the ABCA6 gene is composed of 38 exons which extend across a region of 62 kb size on chromosome 17q24.2. Based on its structural features and its cholesterol-responsive regulation ABCA6 is potentially involved in macrophage lipid homeostasis. PMID- 11478799 TI - Differential effects of IL-2 and IL-15 on expression of IL-2 receptor alpha. AB - IL-2 and IL-15 have overlapping functions since they share the IL-2Rbetagamma receptor complex. However, each cytokine has a private alpha receptor namely IL 2Ralpha for IL-2 and IL-15Ralpha for IL-15. As a consequence the effects of the two cytokines may differ. We describe the differential effects of the two cytokines regarding the induction of cell surface expression of the IL-2Ralpha subunit on YT-l cells. Both cytokines induced transcription of the IL-2Ralpha gene. Furthermore translation of IL-2Ralpha leading to intracellular expression of the receptor was observed following either IL-2 or IL-15 addition. However, only IL-15 was associated with the induction of cell surface expression of IL 2Ralpha. With IL-2 there appears to be an impediment to the translocation of IL 2Ralpha to the cell membrane. Since surface expression of IL-2Ralpha is a key element in the formation of the high affinity IL-2 receptor, translocation of IL 2Ralpha to the membrane represents another level of control of the immune response in addition to regulation of IL-2Ralpha transcription and translation. PMID- 11478800 TI - Evidence as a HIV-1 self-defense vaccine of cyclic chimeric dodecapeptide warped from undecapeptidyl arch of extracellular loop 2 in both CCR5 and CXCR4. AB - Novel conformation-specific antibodies were raised against a cyclic chimeric dodecapeptidyl multiple antigen peptide (cCD-MAP) constructed with a spacer-armed Gly-Asp dipeptide and two pentapeptides (S(169)-Q(170)-K(171)-E(172)-G(173) of CCR5 and E(179)-A(180)-D(181)-D(182)-R(183) of CXCR4) which are components of the undecapeptidyl arch (UPA: from R(168) to C(178) in CCR5, from N(176) to C(186) in CXCR4) of extracellular loop 2 (ECL2) in chemokine receptors (CCR5 and CXCR4). Of the antibodies raised, one monoclonal antibody, CPMAb-I (IgMkappa), reacted with cCD-MAP, but not with the linear chimeric dodecapeptide-MAP. The antibody reacted with the cells separately expressing CCR5 or CXCR4, but not with those not expressing the coreceptors. Moreover, the antibody markedly suppressed infection by X4, R5, or R5X4 virus in a dose-dependent manner in a new phenotypic assay for drug susceptibility of HIV-1 using CCR5-expressing Hela/CD4(+) cell clone 1-10 (MAGIC-5). Moreover, CPMAb-I interfered with LAV-1(BRU) infection (m.o.i. = 0.01) of Molt4#8 cells cocultured with CPMAb-I-producing hybridoma in the transwell, and significantly interfered with neither chemotaxis nor calcium influx induced with stromal cell-derived factor 1 alpha (SDF-1alpha). Thus, the antibody raised against the cCD-MAP provides powerful protection or defense against HIV-1 infection. We therefore propose the cCD-MAP or its derivative immunogen as a novel candidate for an HIV-1 coreceptor-based self-defense vaccine. PMID- 11478801 TI - A phenylarsine oxide-binding protein of neutrophil cytosol, which belongs to the S100 family, potentiates NADPH oxidase activation. AB - By photoaffinity labeling with a tritiated azido derivative of phenylarsine oxide (PAO), 4[N-(4-azido-2-nitrophenyl)amino-[(3)H]acetamido]phenylarsine oxide ([(3)H]azidoPAO), we demonstrate that PAO binds selectively to the S100 A8/A9 complex of bovine neutrophil cytosol (previously known as p7/p23, homologous to the MRP-8/MRP-14 complex of human phagocytes). Using a semirecombinant cell free assay of oxidase activation and the determination of oxidase activity by the production of the superoxide anion O(-)(2), we found that the PAO binding protein (p7/p23) was able to potentiate the activation of NADH oxidase and that this effect was synergized by PAO. The p7/p23 protein complex of bovine neutrophils can therefore be considered as a positive regulator of NADPH oxidase activation in neutrophils. PMID- 11478802 TI - Sequence analysis of the human kallikrein gene locus identifies a unique polymorphic minisatellite element. AB - Minisatellites are repetitive sequences of DNA that are present throughout the genome. Although the origin and function of these minisatellites is still unknown, they found clinical applications as markers of many diseases, including cancer. Also, they are useful tools for DNA fingerprinting and linkage analysis. Kallikreins are serine proteases that appear to be involved in many diseases including brain disorders and malignancy. We have recently characterized the human kallikrein gene locus on chromosome 19q13.4, which includes 15 kallikrein genes. In this study, we examined the kallikrein locus ( approximately 300 Kb) for all known repeat elements. About 50% of this genomic area is occupied by different repeat elements. We also identified unique minisatellite elements that are restricted to chromosome 19q13. Ten clusters of these minisatellites are distributed along the locus on either DNA strand. The clusters are located in the promoters and enhancers of genes, in introns, and in untranslated regions of the mRNA. Analysis of these elements indicates that they are polymorphic, thus they can be useful in linkage analysis and DNA fingerprinting. Our preliminary results indicate also that the distribution of the different alleles of these minisatellites might be associated with malignancy. PMID- 11478803 TI - The HGF/SF-induced phosphorylation of paxillin, matrix adhesion, and invasion of prostate cancer cells were suppressed by NK4, an HGF/SF variant. AB - Hepatocyte growth factor/scatter factor (HGF/SF) plays a crucial role in cancer cell migration, matrix adhesion, invasion, and angiogenesis, via the phosphorylation of the c-met tyrosine kinase. This study examined the ability of NK4, a recently discovered HGF/SF variant, to inhibit the influence of HGF/SF on cell-matrix interaction, paxillin phosphorylation, and invasion of prostate cancer cells. HGF/SF was shown to dramatically enhance tumour cell motility, invasion, cell-matrix adhesion, together with an increase in the degree of paxillin phosphorylation and formation of focal adhesion complexes. However, these HGF/SF-induced effects were suppressed by the presence of NK4. NK4 effectively inhibited the degree of HGF/SF-induced paxillin phosphorylation and matrix adhesion. As a consequence, the matrix invasion of these prostate cancer cells was also suppressed by NK4. In conclusion, this study shows that these HGF/SF-enhanced events, which are critical steps in metastasis, can be inhibited through the addition of NK4, thus warranting further in vivo studies on the implication of NK4 as a potential antimetastasis agent in prostate cancer. PMID- 11478804 TI - A novel Xenopus acetyltransferase with a dynamic expression in early development. AB - We have isolated a novel acetyltransferase from Xenopus laevis, named Xat-1. Xat 1 cDNA encodes a predicted protein of 846 amino acids that contains tetratricopeptide repeat (TPR) domains mediating protein-protein interactions and a bipartite nuclear localization signal (NLS). Its apparent molecular mass of 98.8 kDa was determined by SDS-PAGE analysis of Xat-1 recombinant protein in vitro translated in rabbit reticulocyte lysate. Xat-1 is homologous to N-terminal acetyltransferase 1 (NAT1), a gene that was originally discovered in yeast. Furthermore, it has many orthologs from human, mouse, Drosophila, C. elegans, and even Arabidopsis, thereby suggesting that these constitute a novel acetyltransferase family whose functions have been not examined. Xat-1 transcripts are expressed at relatively constant levels throughout early embryonic stages. They also exhibit dynamic expression pattern in brain, somites, branchial arches, pronephros, and otic vesicles. PMID- 11478805 TI - Characterization of the rplB gene from Streptomyces collinus and its protein product by mass spectrometry. AB - Ribosomal protein L2 is the largest protein components of 50S subunits. The protein is implicated in peptidyl transferase activity and binds to functionally important domains of 23S rRNA. The rplB gene, which codes for ribosomal protein L2 was cloned from Streptomyces collinus. The gene rplB was isolated from BamHI fragment (3.0 kb) of chromosomal DNA possessing two partial and four complete ORF's in the order from 5' to 3': rplC, rplD, rplW, rplB, rpsS, and rplV. The gene organization corresponds to the S10 operon. Gene rplB (834 bp) encodes a polypeptide chain of 278 amino acids. The molecular mass calculated from genomic structure is 30.5 kDa and pI 11.87. Protein L2 is rich in positively charged amino acids (Arg 36, Lys 20, and His 11). N-terminal domain possesses topology similar to the oligonucleotide/oligosaccharide binding OB folds. The availability of genome sequence makes it possible to identify L2 protein by mass spectrometry, moreover it facilitates the characterization of its potential posttranslational modifications. To confirm the protein sequence derived from the rplB gene the tryptic peptides of L2 were analyzed by mass spectrometric techniques. The obtained data matched exactly with the results of DNA sequencing. PMID- 11478806 TI - Chromogenic substrates of bovine beta-trypsin: the influence of an amino acid residue in P1 position on their interaction with the enzyme. AB - The Cucurbita maxima trypsin inhibitor CMTI-III molecule was used as a vehicle to design and synthesize a series of trypsin chromogenic substrates modified in position P1: Ac-Ala-Val-Abu-Pro-X-pNA, where X = Orn, Lys, Arg, Har, Arg(NO(2)), Cit, Hci, Phe(p-CN), Phe(p-NH(2)); pNA = p-nitroanilide. The most active compounds (as determined by specificity constant k(cat)/K(m)) were peptides with the Arg and Lys residues in the position discussed. Changes in the length and the decrease of the positive charge of the amino acid residue side chain in position P(1) resulted in the decrease or loss of the affinity towards bovine beta trypsin. Among peptides containing amino acid residues with uncharged side chains in position P1, only one with p-cyano-l-Phe revealed activity. These results correspond well with trypsin inhibitory activity of CMTI-III analogues modified in the equivalent position, indicating the same type of interaction between position P1 of the substrate or inhibitor and S1 site specificity of trypsin. PMID- 11478807 TI - Apoptosis of lymphocytes in the presence of Cr(V) complexes: role in Cr(VI) induced toxicity. AB - Cr(VI) compounds have been declared as a potent occupational carcinogen by IARC (1990) through epidemiological studies among workers in chrome plating, stainless steel, and pigment industries. Studies relating to the role of intermediate oxidation states such as Cr(V) and Cr(IV) in Cr(VI)-induced carcinogenicity are gaining importance. In this study, issues relating to toxicity elicited by Cr(V) have been addressed and comparisons made with those relating to Cr(VI) employing human peripheral blood lymphocytes. Lymphocytes have been isolated from heparinized blood by Ficoll-Hypaque density gradient centrifugation and exposed to Cr(V) complexes viz. sodium bis(2-ethyl-2-hydroxybutyrato)oxochromate(V), Na[Cr(V)O(ehba)(2)], 1 and sodium bis(2-hydroxy-2-methylbutyrato)oxochromate(V), Na[Cr(V)O(hmba)(2)], 2 and Cr(VI). The phytohemagglutinin (PHA)-induced proliferation of lymphocytes has been found to be inhibited by the two complexes of Cr(V) and chromate Cr(VI) in a time- and concentration-dependent manner. Viability of cells decreases in the presence of Cr(V). Apoptosis appears to be the mode of cell death in the presence of both Cr(V) and Cr(VI). Pretreatment of cells with antioxidants before exposure to chromium(V) complexes reverse apoptosis partially. Possibility for the formation and implication of reactive oxygen species in Cr(V)-induced apoptosis of human lymphocyte cells has been indicated in this investigation. The intermediates of Cr(V) and radical species in the biotoxic pathways elicited by Cr(VI) seems feasible. PMID- 11478808 TI - TR4 orphan receptor represses the human steroid 21-hydroxylase gene expression through the monomeric AGGTCA motif. AB - The human TR4 orphan receptor (TR4) is a member of the nuclear receptor superfamily. It functions as a transcriptional factor which regulates and controls many important physiological functions. It has been documented that TR4 may bind as a homodimer to a DNA response element containing two direct repeats of the AGGTCA consensus motif. Surprisingly, our data reveal that the expression of the human steroid 21-hydroxylase (21-OHase) gene could be repressed by TR4 via the monomeric AGGTCA motif (-228TR4RE) at its 5' flanking region (nucleotide numbers 1431-1444, 5'-GGAAAAAGGTCAGG-3'). Electrophoretic mobility shift assay showed specific binding with a dissociation constant of 0.4 nM between TR4 and the monomeric -288TR4RE motif. However, TR4 does not form heterodimers with either retinoid X receptor alpha or SHP (short heterodimer partner) orphan receptor. Additionally, both dual-luciferase and chloramphenicol acetyltransferase assays demonstrated that TR4 can function as a repressor via the -228TR4RE of the 21-OHase gene. In conclusion, our data suggest that TR4 may bind to a monomeric DNA response element and play an important role in the suppression of the 21-OHase gene expression. PMID- 11478809 TI - The paralemmin protein family: identification of paralemmin-2, an isoform differentially spliced to AKAP2/AKAP-KL, and of palmdelphin, a more distant cytosolic relative. AB - Paralemmin is a protein implicated in plasma membrane dynamics. Here we describe the identification of two new paralemmin-related proteins. A partial paralemmin homolog, palmdelphin, is predominantly cytosolic, unlike paralemmin which is lipid-anchored to the plasma membrane through a C-terminal CaaX motif. We have mapped the mouse palmdelphin gene to distal chromosome 3 between Amy2 and Abcd3, in a region homologous to human chromosome 1p22-p21 where the human palmdelphin gene is located. We have also identified a second paralemmin isoform, paralemmin 2. It is expressed from a gene on human chromosome 9q31-q33 which ends only 33 kb upstream of the gene encoding the protein kinase A-binding protein,AKAP2/AKAP-KL. The closely adjacent paralemmin-2 and AKAP2 genes are functionally linked in a very unusual manner. Chimeric mRNAs are expressed, apparently by RNA readthrough and differential splicing, that encode natural fusion proteins in which either the N-terminal coiled-coil region or nearly the complete sequence of paralemmin-2 except its C-terminal CaaX motif is fused to AKAP2/AKAP-KL. The N-terminal coiled coil region is conserved in paralemmin-1, paralemmin-2/AKAP2, palmdelphin and a fourth, uncharacterized gene, suggesting that it is a modular functional domain. PMID- 11478810 TI - Tissue distribution and subcellular localization of a variant form of the human ST2 gene product, ST2V. AB - The human ST2 gene has been known to encode three splice variants; namely, a soluble secreted form of ST2, a transmembrane form of ST2L, and ST2V of undetermined localization. Therefore, analysis of tissue distribution and subcellular localization of ST2V is important to elucidate functional relationships among the three splice variants of the human ST2 gene. RT-PCR procedure revealed that ST2V is predominantly expressed in the stomach, small intestine, and colon. Transfection of ST2V cDNA into COS7 cells in the presence of [(35)S] methionine and cysteine produced radiolabeled 40 kDa protein, which is recognized by specific monoclonal antibody against human ST2. Subcellular fractionation analysis showed that ST2V protein was distributed in the insoluble fraction of the cell lysate. Finally, ST2V protein was detected on the plasma membrane of COS7 cells, which had been transfected with ST2V cDNA, by confocal laser microscopic analysis. These findings taken together, indicate that ST2V protein localizes on the plasma membrane, suggesting its possible role in modification of the ST2L-signaling pathways. PMID- 11478811 TI - Untoward effects of rapid-acting antihypertensive agents. AB - Cerebral haemorrhage remains a major cause of hypertensive cerebral mortality. Lowering of raised blood pressure is therefore crucial in the clinical management of hypertensive disorders of pregnancy. This article reviews rapid-acting agents employed in life-threatening situations. PMID- 11478812 TI - Anaesthesia and analgesia for the critically ill parturient. AB - Anaesthetic and analgesic techniques in the critically ill are determined largely by the nature of the presenting illness. The commonest conditions likely to present as life-threatening emergencies are pre-eclampsia, obstetric haemorrhage, cardiac disease and severe sepsis. Issues dictating choice of anaesthetic technique are the patient's ability to maintain her airway, coagulation status, intravascular volume and haemodynamic dependence upon sympathetic drive, and requirements for ventilatory support and intensive care. Fetal well-being is an issue in the antepartum period, uteroplacental blood flow should be maintained and hypotension avoided. Maternal survival takes priority, however, and occasionally general anaesthetic techniques must be used which lead to neonatal respiratory depression and requirement for ventilatory support. Anaesthesia itself is associated with known hazards. The risks of each technique must be balanced against possible benefits in the context of the presenting illness. PMID- 11478813 TI - Management of the critically ill cardiac patient. AB - The decline in rheumatic fever has made heart disease in pregnancy an uncommon problem in the developed world but it remains an important cause of maternal morbidity and mortality in developing countries. Pregnancy is particularly dangerous in the presence of cyanotic congenital heart disease, Eisenmenger's syndrome, primary pulmonary hypertension, Marfan's syndrome, dilated cardiomyopathy and significant mitral stenosis. Severe stenosis is often complicated by pulmonary hypertension and atrial fibrillation. Maternal disease status should be determined using echocardiography to define cardiac anatomy, assess ventricular function and estimate intracardiac pressure gradients. Patients in the New York Heart Association functional classes 1 and 2 generally have a favourable outcome. Closed mitral commissurotomy is safe and effective in relieving stenosis across the mitral valve in selected patients. More recently the technique of percutaneous balloon mitral valvotomy has successfully been used in the treatment of mitral stenosis. Termination of pregnancy is advised in patients with severe pulmonary hypertension, including Eisenmenger's syndrome. PMID- 11478814 TI - Ventilation and the critically ill parturient. AB - Positive-pressure ventilation is the keystone in the management of pulmonary dysfunction in the critically ill. An increased understanding of both the benefits and hazards has led to a general consensus regarding the optimal techniques to ensure adequate gas exchange. Unfortunately, the same cannot be said for ventilation terminology which, due to a lack of standardization, lends itself to confusion. Pulmonary dysfunction in the parturient may arise from thoracic or extra-thoracic pathologies but both may be defined as acute lung injury. In its most severe form this constitutes acute respiratory distress syndrome. Acute lung injury results in reduced lung compliance and a marked decrease in the volume of functional lung. Ventilation strategies are now designed to recruit as much available lung tissue as possible while simultaneously minimizing the injurious effects of alveolar over-distension. Upon resolution of the underlying pathology mechanical ventilation may be withdrawn. Recent evidence suggests that this final stage need not be protracted, and if certain criteria are fulfilled, rapid weaning is feasible. PMID- 11478815 TI - Arterial embolization for haemorrhage in the obstetric patient. AB - Arterial embolization is a safe and effective treatment for persistent post partum haemorrhage that is unresponsive to conservative management. Embolization should be the treatment of choice in these patients provided that suitable radiological skills and equipment are available. Embolization is potentially useful in patients with antepartum haemorrhage in the last trimester or in patients at high risk for antepartum haemorrhage. PMID- 11478816 TI - Care of the critically ill parturient: oliguria and renal failure. AB - The incidence of acute renal failure in pregnancy has decreased. This decrease is less marked in developing countries in which resources are more scarce. The clinical diagnosis of acute renal failure is crude due to the variability of clinical signs and the late occurrence of basic biochemical abnormalities. Obstetric and gynaecological diseases are found among the traditional pre-renal, intra-renal and post-renal causes of acute renal failure. The cornerstone of management is the identification of high-risk cases and the prevention of acute renal failure by maintaining intravascular volume. The evidence for the efficacy of other prophylactic medical interventions, such as the use of loop diuretics, mannitol, low-dose dopamine and others, is poor. Management of established acute renal failure includes restoration of intravascular volume, treatment of any reversible causes, especially pregnancy complications such as pre-eclampsia, strict fluid balance and correction of any electrolyte abnormality or metabolic acidosis. Dialysis is a supportive measure until the kidneys recover. PMID- 11478817 TI - Management of venous thromboembolism in pregnancy. AB - Pulmonary thromboembolism (PTE) is the major cause of maternal death in the UK, with recent trends showing an increase in the numbers of deaths. Underlying PTE is the problem of deep venous thrombosis (DVT). An appreciation of risk factors, particularly, thrombophilia, and signs or symptoms suggestive of thromboembolism, coupled with objective diagnosis and treatment should reduce mortality and morbidity. There are particular considerations with regard to the management of thrombosis in pregnancy, especially the use of anticoagulants. Low-molecular weight heparins are now replacing unfractionated heparin for the treatment of DVT and PTE in pregnancy. PMID- 11478818 TI - Haemodynamic, invasive and echocardiographic monitoring in the hypertensive parturient. AB - AIM: To determine the clinical usefulness of invasive and non-invasive haemodynamic assessment in pre-eclampsia. METHOD: A systematic review of the literature was undertaken, using a MEDLINE electronic search using a combination of MESH headings and textwords. Over 1500 abstracts were perused; we obtained 156 full papers that were related to the subject matter. Of the full papers, 55 yielded relevant information. Hand-searching the reference lists of the retrieved papers completed the search. RESULTS: There are no data from randomized controlled clinical trials illustrating the clinical usefulness of pulmonary artery catheters or echocardiographic techniques in hypertensive pregnancy. There are a wealth of data illustrating the haemodynamic profiles of both untreated and treated pre-eclamptic women. Data are also available comparing right heart and left heart filling pressures, demonstrating a relatively poor correlation between the two values. The clinical impact of either measurement is unclear. Data are available illustrating the correlation between echocardiographic techniques and pulmonary artery catheterization. PMID- 11478819 TI - Disseminated intravascular coagulation. AB - Healthy pregnancy is accompanied by changes in the haemostatic system which convert it into a hypercoagulable state vulnerable to a spectrum of disorders ranging from venous thromboembolism to disseminated intravascular coagulation (DIC). This latter is always a secondary phenomenon triggered by specific disorders such as abruptio placentae and amniotic fluid embolism due to release of thromboplastin intravascularly or endothelial damage resulting from pre eclampsia and sepsis. In modern obstetric practice the most common cause is haemorrhagic shock with delay in resuscitation leading to endothelial damage. The initial management of massive obstetric haemorrhage is the same whether associated with coagulopathy initially or not. Low-grade DIC, associated with pre eclampsia, is monitored haematologically by serial platelet counts and serum fibrin degradation products (FDPs). Supportive measures and removal of the triggering mechanism are the key to successful management. Outcome depends primarily on our ability to deal with the trigger and not on direct attempts to correct the coagulation deficit. PMID- 11478820 TI - Hypovolaemic shock. AB - Measured blood loss up to 1000 ml is well tolerated by healthy pregnant women. This is partly due to physiological increases in plasma volume and red cell mass during pregnancy. Nevertheless, hypovolaemic shock is a major cause of maternal mortality. Management requires teamwork, co-ordination, speed and adequate facilities to be life-saving. The first priority is rapid fluid replacement. Evidence from randomized trials has established that crystalloids are the fluids of choice over colloids and particularly albumen, which was associated with increased mortality. Rapid access to blood or blood products for transfusion is necessary, as well as laboratory back-up. Further management includes accurate assessment of the site of bleeding; control of the bleeding; diagnosis and management of the underlying condition; supportive therapy; and monitoring of the clinical, haematological and biochemical response to treatment. Bedside diagnostic ultrasound has several applications in the evaluation of obstetric hypovolaemic shock. PMID- 11478824 TI - Nutrition support teams: dissociated ions or the yeast in the loaf? PMID- 11478825 TI - Nutritional support in chronic renal failure: systematic review. AB - BACKGROUND AND AIMS: The purpose of this systematic review was to locate and assess in patients with chronic renal disease the quality of scientific evidence to establish graded recommendations based on the efficacy and effectiveness of nutritional support. METHODS: Computerized and online versions of MEDLINE (from 1989 through March 1999) and EMBASE (from 1988 through January 1999) were consulted. The Cochrane Library and the online Healthstar (from 1975) databases were also searched for clinical trials. A total of 593 studies were assessed. Following methodological review (primary reviewer), only 45 studies reviewed met criteria for selection and were analyzed by a group of experts (secondary reviewer). A final consensus was reached between the co-ordinators, experts and methodologists. RESULTS AND CONCLUSIONS: Low and very low-protein diets associated with specific enteral supplements are indicated in most patients with chronic renal disease and in patients with incipient diabetic nephropathy to slow progression of disease and to improve their overall status, contributing to improved survival (grade A recommendation). The use of protein-restricted diets in diabetic nephropathy is only indicated in type I diabetes mellitus (grade A recommendation). An improvement in nutritional parameters was found when specific diets were given in chronic renal disease (grade C recommendation). PMID- 11478826 TI - Soluble fiber reduces the incidence of diarrhea in septic patients receiving total enteral nutrition: a prospective, double-blind, randomized, and controlled trial. AB - BACKGROUND AND AIMS: Attempts to control enteral nutrition associated diarrhea in the critically ill tube-fed patient by implementing feeding formulas enriched with fiber were mostly unsuccessful. Recently, it was shown that enteral feeding containing soluble partially hydrolyzed guar decreased the incidence of diarrhea in a cohort of non-critically ill medicosurgical patients. We investigated whether this type of enteral feed could also influence stool production in patients with severe sepsis, a population at risk for developing diarrhea. METHODS: The study was double-blind. Patients with severe sepsis and septic shock were consecutively enrolled and at random received either an enteral formula supplemented with 22 g/l partially hydrolyzed guar or an isocaloric isonitrogenous control feed without fiber. All patients were mechanically ventilated and treated with catecholamines and antibiotics. Enteral feeding was provided through a nasogastric tube for a minimum of 6 days. A semiquantitative score based on stool volume and consistency was used for daily assessment of diarrhea. RESULTS: 25 patients fulfilled the criteria for data analysis. Soluble fiber was administered in 13 of them. The two groups were well-matched for gender, age, disease severity, cause of sepsis, laboratory parameters, total feeding days and time to reach nutritional goals. The mean frequency of diarrhea days was significantly lower in patients receiving fiber than in those on standard alimentation (8.8+/-10.0 % vs 32.0+/-15.3 %; P=0.001). The whole group of fiber-fed patients had less days with diarrhea per total feeding days (16/148 days (10.8%) vs 46/146 days (31.5%); P<0.001) and a lower mean diarrhea score (4.8+/-6.4 vs 9.4+/-10.2; P<0.001). The type of enteral diet did not influence sepsis-related mortality and duration of stay in the intensive care unit. CONCLUSION: Total enteral nutrition supplemented with soluble fiber is beneficial in reducing the incidence of diarrhea in tube-fed full-resuscitated and mechanically ventilated septic patients. PMID- 11478827 TI - Does the supplementation of the formula with fibre increase the risk of gastro oesophageal reflux during enteral nutrition? A human study. AB - BACKGROUND & AIMS: Fibre-supplements in enteral feeding could increase the risk of gastro-oesophageal reflux (GOR). The aim of this study was to assess in humans the physiological effects of the supplementation of the enteral diet with different types of dietary fibre on gastro-oesophageal reflux episodes, gastric acidity and gastric emptying. METHODS: pH profiles were compared in 12 healthy volunteers between three different formula (500 kcal, 250 mL x h-1) delivered in a random order and containing either no fibre, either soy polysaccharide fibre only or mixed fibre from pea and inuline. Enteral diets were instilled through a nasogastric tube. Oesophageal and gastric pH recordings were combined with the ultrasound measurement of gastric antral area during the infusion. RESULTS: More GOR were observed with a fibre-free diet (median 4, range 1-10) than with a mixed (median 1.5, range 0-5) (P=0.04) or soy polysaccharide fibre (median 1.5, range 0 5) (P=0.04) diet. The median duration of GOR was longer with the mixed fibre (median 3.6, range 1.8-7.2) than with the fibre-free diet (mean 1.8 min, range 1 3.6) (P<0.05). The number of GOR episodes lasting more than 5 min, the duration of the longest GOR and the percent of time under pH 4 were not significantly different with the three diets. The intragastric pH profile and the ultrasound antral area were not different with all three diets. CONCLUSIONS: Addition of fibre to the enteral formula had limited effects on the onset of GOR episodes. It decreased the number of GOR but increased their duration. These effect were more pronounced with the formula containing soluble fibre. At variance, the addition of fibre had no significant effect on gastric emptying and gastric acid secretion. PMID- 11478828 TI - Cytokine-mediated inhibition of ketogenesis is unrelated to nitric oxide or protein synthesis. AB - Cytokines play an important role in the lipid disturbances commonly associated with sepsis. Ketogenesis is inhibited during sepsis, and tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) have been suggested to mediate this impairment, irrespective of the ketogenic substrate (fatty acid or branched chain ketoacid). However, the underlying mechanism of cytokine action is still unknown. First we investigated the possible role of the induction of nitric oxide (NO) synthesis, using rat hepatocyte monolayers. Hepatocytes were incubated for 6 h, with either alpha -ketoisocaproate (KIC) (1 mM) or oleic acid (0.5 mM) in the presence or absence of TNF alpha (25 microg/L) and IL-6 (15 microg/L). In some experiments, cells were incubated with NO synthase (NOS) inhibitors. The ketone body (beta -hydroxybutyrate and acetoacetate) production and nitrite production were measured in the incubation medium. Our results indicated no involvement of nitric oxide in the inhibitory action of cytokines on ketogenesis. Secondly, we showed that cycloheximide (10(-4)M) did not counteract the cytokine-mediated ketogenesis decrease; hence, the effects of cytokines on ketogenesis are not protein synthesis-dependent. The cytokine-mediated inhibition of ketogenesis is therefore unrelated to either NO production or protein synthesis. PMID- 11478829 TI - Effect of glutamine in short-bowel syndrome. AB - BACKGROUND AND AIMS: Animal studies have reported positive effects of glutamine on intestinal absorption and morphology; human studies have been less convincing. The aim of this study was to evaluate the effects of glutamine and diet on intestinal morphology, motility, and absorption. METHODS: A randomized, double blind, placebo-controlled crossover study in 8 patients with short-bowel on a high carbohydrate, low fat (HCLF) diet, was performed. Active treatment was oral glutamine (0.45 g kg(-1)day(-1)) for eight weeks. Intestinal morphology was evaluated by light microscopy. Gastrointestinal transit was measured by dual gamma camera scintigraphy. D-xylose and fecal fat collection was used to evaluate intestinal absorption. Results of active treatment versus placebo were compared by the signed-rank test. RESULTS: Morphology analysis, reported as median active treatment versus placebo, was villus height: 0.48 mm versus 0.50 mm, P=1.0, and crypt depth: 0.11 mm versus 0.10 mm, P=0.469. Percent D-xylose absorption, reported as median active treatment versus placebo, was 7% versus 10.5%, P=0.109. There was not a significant difference in wet weight or fat absorption compared to placebo, P>0.05. Likewise, gastrointestinal transit was not different compared to placebo. CONCLUSIONS: The results of this controlled study would support that 8 weeks of treatment with oral glutamine and a HCLF diet does not significantly improve intestinal morphology, gastrointestinal transit, D-xylose absorption and stool losses in short bowel patients. PMID- 11478830 TI - Effect of fasting on rat duodenal and jejunal microvilli. AB - Duodenal and jejunal ultrastructural morphology and total serum alkaline phosphatase (ALP) activity were investigated in Wistar rats submitted to normal feeding, a 24 h fasting without or with access to sawdust or expanded polystyrene, and a 22 h fasting followed by a 2 h refeeding. Ultrastructural observation of duodenal and jejunal villous tips showed the occurrence of degenerating epithelial cells that were much more frequent in the epithelium of fasted rats. Refeeding and ingestion of sawdust or expanded polystyrene decreased the mean percentage of degenerating cells in the duodenum but not in the jejunum in comparison to plain fasted animals. Normal feeding produced the highest value of serum ALP activity. Refeeding as well as ingestion of sawdust or expanded polystyrene increased serum ALP activity values from those of plain fasted state: the difference being significant only for the refed group. It is concluded that: a) feeding, for both duodenum and jejunum, and refeeding and mechanical stimulation, just for the duodenum, increased proliferation, decreased apoptosis or facilitated desquamation of the epithelium; b) ALP increase in serum was induced by feeding and refeeding. PMID- 11478831 TI - Patterns of prescribing of nutritional supplements in the United Kingdom. AB - BACKGROUND AND AIMS: A large number of prescriptions are issued for nutritional supplements under British National Formulary classifications 9.4.1 (foods for special diets) and 9.4.2 (enteral feeds), but little is known about the characteristics of the patients who receive them. We used the General Practice Research Database to examine patterns of prescribing of these supplements. METHODS: We selected patients who had been prescribed supplements under classifications 9.4.1 and 9.4.2 during 1996-1997. Descriptive statistics were used to examine how prescribing varied. RESULTS: 28644 patients received prescriptions during 1996-1997. Among the 27413 (96%) patients prescribed supplements for oral use, 14750 received supplements for enteral nutrition alone, 8122 received supplements for special diets alone and 4541 had both types of supplement. 51% of patients receiving supplements for special diets were <18 years. The commonest diagnoses among such children were milk intolerance (24%) and malnutrition (17%). 94% of patients receiving supplements for enteral nutrition were adult, 52% of whom had cancer or cardiovascular disease. Only 4% of patients had weight and height recorded prior to first prescription. CONCLUSIONS: The GPRD provides valuable information on the characteristics of patients prescribed nutritional supplements. But because only limited data are available on their nutritional status prior to supplementation, it is hard to assess whether general practitioners are prescribing these supplements appropriately. PMID- 11478832 TI - Body water compartment measurements: a comparison of bioelectrical impedance analysis with tritium and sodium bromide dilution techniques. AB - BACKGROUND AND AIMS: This study was undertaken to assess the comparability of body water compartment estimates in healthy volunteers using single and dual frequency bioelectrical impedance analysis (BIA) with established reference methods of tritium and NaBr dilution. METHODS: Total body water (TBW) was estimated in 10 healthy volunteers using single frequency (50 kHz) BIA (Bodystat 1500), dual frequency (5 and 200 kHz) BIA (Bodystat Dualscan 2005) and tritium dilution. Extracellular water (ECW) was measured with dual frequency BIA and NaBr dilution. BIA was performed using distal tetrapolar electrodes in the supine position. Venous blood was sampled for measurement of background concentrations of tritium and NaBr using a beta counter and high performance liquid chromatography respectively. 10 ml tritiated water (3.7 MBq) and 50 ml 5% NaBr solution were then injected intravenously and blood samples taken from the opposite arm every 45 min for 4.5 h for estimation of concentrations of tritium and NaBr. RESULTS: There was good correlation (r(2)=0.76) between estimates of ECW using dual frequency BIA and NaBr dilution, with the former overestimating ECW by approximately 1 L. However, this difference varied systematically with body weight. Although TBW measurements obtained by single and dual frequency BIA correlated well with estimates using tritium dilution (r(2)=0.96 and 0.95 respectively), single frequency BIA underestimated TBW by approximately 1 L and dual frequency BIA by approximately 5 L compared to tritium dilution. CONCLUSION: TBW measurements obtained using the single frequency BIA device were more accurate than those obtained using the dual frequency BIA device. Dual frequency BIA provided a reasonably accurate estimate of ECW. PMID- 11478833 TI - Prospective survey of parenteral nutrition in Switzerland: a three-year nation wide survey. AB - BACKGROUND AND AIMS: The goals of this national survey were to determine the current PN practices and admixture formulations used in Switzerland. METHODS: During three years, an annual questionnaire was sent to all heads of Swiss hospital pharmacies. RESULTS: 92% of Swiss hospitals with a full-time pharmacist participated. Different PN systems were commonly used for adult patients: 2 commercial formulas in 2 or 3 compartments bags, 2 commercial formulas with/without lipid, 3 formulas compounded by the hospital pharmacy. For hospitalized adults, 83% of PN bags were administered as commercial multicompartment bags. The compounding of individualized PN admixtures takes place primarily in pharmacies of medium to large size hospitals. For pediatric PN, hospital compounding is routine because of individualized PN compositions and absence of commercially available standardized admixtures. Long-term home-PN was mostly delivered by hospital pharmacies (57%) or by private nutrition support home delivery services (37%). Most PN formula compositions complied with European guidelines and represented 2.6+/-2.0% of the hospital drug budget. Multi disciplinary nutritional support teams were present in 52% of hospitals. CONCLUSION: In Switzerland, most PN for hospitalized adults were administered as commercial multi-compartment bags. The compounding of individualized PN admixtures were still important for pediatric patients and long-term home-PN. PMID- 11478834 TI - Effect of eicosapentaenoic acid (EPA) on tight junction permeability in intestinal monolayer cells. AB - The purpose of this study is to evaluate the effect of C18 and C20 long chain fatty acids on tight junction permeability in a model of intestinal epithelium. METHODS: Confluent Caco-2 cells on porous filters with double chamber system were used to measure fluorescein sulfonic acid (FS) permeability and transepithelial electrical resistance (TEER). Lactate dehydrogenase release and ultrastructure were evaluated. Effect of 200 microM eicosapentaenoic acid (EPA, C20:5 n-3), arachidonic acid (AA, C20: 4 n-6), alpha-linoleic acid (ALA, C18: 3 n-3), linoleic acid (LA, C18: 2 n-6), or oleic acid (OA, C18: 1 n-9) enrichment in the culture medium during 24 hours were compared. The effect of the cyclooxygenase inhibitor, indomethacin, lipoxygenase inhibitors, NDGA or AA861, and antioxidant, BHT, was evaluated as a mechanism to change tight junction permeability. RESULTS: Caco-2 cells formed polarized columnar epithelial cells with densely packed microvilli and well developed junctional complexes. Addition of EPA enhanced FS permeability to 3.0+/-1.6-fold and lowered TEER to 0.59+/-1.2-fold vs. control with concentration dependency without cell injury (P<0.01-0.05). OA, AA or LA did not change, but ALA enhanced tight junction permeability. Indomethacin and AA861 normalized the changes mediated by EPA. CONCLUSIONS: EPA affects tight junction permeability in intestinal monolayer cells specifically and concentration dependently via cyclooxygenase and lipoxygenase products. PMID- 11478835 TI - Towards implementation of optimum nutrition and better clinical nutrition support. AB - Clinical Nutrition Support--defined as nutrition for hospitalized patients suffering from metabolic stress--plays a limited role in the therapeutic routine of the physician. This is not surprising as most research in the field of clinical nutrition is disappointing with regard to the objective outcomes: morbidity and mortality. These reflections advocate a more 'pharmaceutical approach' to nutrition in order to perform more proper studies on the potential effectiveness of this treatment modality. To provide all patients in the Academic Medical Centre (AMC) in Amsterdam, The Netherlands, with optimum clinical nutrition support, a Nutrition Support Team (NST) was established in 1996. This NST is coaching the dieticians and physicians in the AMC regarding clinical nutrition support. In practice this coaching consists of providing clear guidelines on what is supposed to be optimum nutrition, a basic course in parenteral nutrition and further continuous education. The concept of optimum nutrition is spread by the NST through various ways of education, both nationally and internationally. For adults, optimum nutrition is defined as the amount of protein, that stimulates whole body protein synthesis maximally (1.7 g/kg actual body weight) and covers anabolic energy need (35 kcal/kg actual body weight). The dietician is considered to be the expert in the field of optimum nutrition by oral, enteral or parenteral route. The Dietetic Department has increased its influence in the care of the patient by placing nutritional status and care on the chart of the patient's treatment. To provide optimal Nutrition Support for children and severe ill patients (Intensive care department) specialized teams were started which were co-ordinated by the central NST. The central NST has a co ordinating and educating role, while the Specialized Nutrition Support Teams (Specialized NST) construct guidelines, undertake research and provide continuous optimum nutrition care. PMID- 11478836 TI - Organizational aspects of starting and running an effective nutritional support service. AB - Setting up and running a Clinical Nutritional Support Team can be challenging and is time consuming. Team members have to be motivated, committed and persistent. High quality working relationships are fundamentally important within the team, between teams and with other healthcare professionals. Good communication, careful planning, sensitive implementation and robust monitoring are the cornerstones of a successful service. Looking forward as well as back will help the team to maintain and improve its position in an ever-changing environment. While nutritional support is everyone's business, an effective multiprofessional Clinical Nutritional Support Team is the best way to ensure that patients receive appropriate and timely treatment. It is also wise to remember that a successful Clinical Nutritional Support Team will be as committed to its own development as it is to the care of the patients referred to it. PMID- 11478837 TI - Liver transplantation for type Ib glycogenosis with reversal of cyclic neutropenia. AB - We describe a case of glycogen storage disease type Ib in 32-year old male patient with poor metabolic control in spite of medical and nutritional management and the use of recombinant granulocyte stimulating factor. Because of this, liver transplantation was considered as a definitive treatment. We comment on the metabolic results of liver transplantation performed, with reversal of hypoglycemia, hyperuricemia, hypertriglyceridemia and cyclic neutropenia, all of which persist 4 years post-transplant. In view of this case, we believe that liver transplantation is a feasible option to consider in patients with type Ib glycogenosis as a definitive therapeutic procedure. PMID- 11478838 TI - Nuclear envelope and chromatin compositional differences comparing undifferentiated and retinoic acid- and phorbol ester-treated HL-60 cells. AB - The human leukemic cell line (HL-60) can be induced to differentiate in vitro to granulocytic form with retinoic acid (RA), or to monocytic/macrophage form with phorbol ester (TPA). The granulocytic form acquires nuclear lobulation, nuclear envelope-limited chromatin sheets (ELCS), and cytoskeletal polarization, none of which are acquired following treatment with TPA. Immunoblotting analyses and capillary zone electrophoresis demonstrated that following RA treatment: lamins A/C and B1, and vimentin decreased to negligible amounts; LAP2 beta, lamin B2 and emerin remained essentially unchanged; lamin B receptor (LBR) increased markedly; histone subtypes H1.4 and 1.5 exhibited dephosphorylation. Following TPA treatment: lamins A/C and B1, B2 and vimentin increased in amount; LAP2 beta and emerin remained essentially unchanged; LBR increased markedly; histone subtypes H1.4 and 1.5 exhibited dephosphorylation. Emerin, which was cytoplasmic in undifferentiated or granulocytic cells, localized into the nuclear envelope following TPA. Normal human granulocytes revealed compositional differences compared to granulocytic forms of HL-60, namely increased vimentin and appearance of histone subtype H1.3. A working hypothesis for nuclear lobulation postulates a combination of: increased nuclear envelope deformability due to lamins A/C and B1 deficiency; an increase in nuclear surface area/volume; an increase in chromatin nuclear envelope interactions. PMID- 11478839 TI - Retinoic acid- and bone morphogenetic protein 4-induced apoptosis in P19 embryonal carcinoma cells requires p27. AB - During development, many cells are specifically eliminated. Therefore, programmed cell death must be understood to fully elucidate embryogenesis. Retinoic acid (RA) and bone morphogenetic protein (BMP) 4 induce rapidly dividing P19 embryonal carcinoma cells to undergo apoptosis. RA alone minimally induces apoptosis, while BMP4 alone induces none. RA and BMP4 exposure also elevates the number of cells in the G1 phase of the cell cycle. Because many cell cycle proteins control both proliferation and apoptosis, we determined the role of these proteins in inducing apoptosis. Although the mRNA levels of cyclins D1 and D2 are reduced in cells undergoing apoptosis, the protein levels are not. In contrast, RA and BMP4 induce the Cdk inhibitor p27. This protein binds Cdk4 in RA- and BMP4-treated cells and inhibits Cdk4-dependent kinase activity. We used p27 antisense oligonucleotides to rescue the P19 cells from RA and BMP4 apoptosis thus proving that p27 is necessary. The Cdk4 substrate, retinoblastoma (Rb) protein, is also induced in apoptotic cells. Consistent with the decreased kinase activity of the apoptotic cells, this Rb protein is hypophosphorylated and presumably active. These data support the hypothesis that RA and BMP4 together induce the p27 protein leading to Rb activation and ultimately apoptosis. PMID- 11478840 TI - Autophagy delays sulindac sulfide-induced apoptosis in the human intestinal colon cancer cell line HT-29. AB - Autophagy is a major catabolic process allowing the renewal of intracellular organelles by which cells maintain their homeostasis. We have previously shown that autophagy is controlled by two transduction pathways mediated by a heterotrimeric Gi3 protein and phosphatidylinositol 3-kinase activities in the human colon cancer cell line HT-29. Here, we show that 3-methyladenine, an inhibitor of autophagy, increases the sensitivity of HT-29 cells to apoptosis induced by sulindac sulfide, a nonsteroidal anti-inflammatory drug which inhibits the cyclooxygenases. Similarly, HT-29 cells overexpressing a GTPase-deficient mutant of the G(alpha i3) protein (Q204L), which have a low rate of autophagy, were more sensitive to sulindac sulfide-induced apoptosis than parental HT-29 cells. In both cell populations we did not observe differences in the expression patterns of COX-2, Bcl-2, Bcl(XL), Bax, and Akt/PKB activity. However, the rate of cytochrome c release was higher in Q204L-overexpressing cells than in HT-29 cells. These results suggest that autophagy could retard apoptosis in colon cancer cells by sequestering mitochondrial death-promoting factors such as cytochrome c. PMID- 11478841 TI - Pharmacological evidence for system-dependent involvement of protein kinase C isoenzymes in phorbol ester-suppressed gap junctional communication. AB - Several phorbol esters are potent activators of protein kinase C. They down regulate gap junctional intercellular communication and induce phosphorylation of connexin43, but the sensitivity and extent of responses vary much between systems. We asked whether the total protein kinase C enzyme activity or the protein kinase C isoenzyme constitution was of importance for such variations. Some fibroblastic culture systems were compared. It was concluded that the total protein kinase C enzyme activity did not determine the sensitivity to phorbol esters. Furthermore, the use of isotype-specific inhibitors of protein kinase C indicated that protein kinase C alpha, delta, and epsilon may be involved to different extents in different fibroblastic systems in the response to phorbol esters. PMID- 11478842 TI - Distinct experimental efficacy of anti-Fas/APO-1/CD95 receptor antibody in human tumors. AB - Ligation of the Fas receptor (FasR) is a key step in apoptosis induction. Using a series of human tumor cells (SNB19, SNB79, 143N2, and SHEP), we observed a distinct efficacy of human anti-FasR antibody with an apparent correlation with Fas cell surface antigen expression. In contrast, all cells studied expressed detectable FasR mRNA transcripts. For all anti-FasR antibody-sensitive tumor cells, we showed a similar efficacy of Mab according to dose fractionation and injection site. We showed that, when injected into nude mice bearing human osteosarcoma 143N2, neuroblastoma SHEP, prostatic cancer PAC120, and the two glioblastomas SNB19 and SNB79, anti-FasR Mab induces significant inhibition of the growth rate of 143N2, SHEP, and PAC120 tumors, but has no efficacy on SNB19 and SNB79 tumors, with a relationship between in vitro and in vivo sensitivity to anti-FasR antibody. Altogether, these results suggest the antitumor potential of anti-FasR antibody in human neoplasms. PMID- 11478843 TI - Inhibition of FGF-2-mediated chemotaxis of murine brain capillary endothelial cells by cyclic RGDfV peptide through blocking the redistribution of c-Src into focal adhesions. AB - The alpha(v)beta(3) integrin is essential for fibroblast growth factor (FGF) induced angiogenesis in vivo. However, the role of this integrin in FGF-2 mediated cellular responses by cultured endothelial cells is largely unknown. Cyclic RGDfV (cRGDfV) peptide is widely used to inhibit the binding of alpha(v)beta(3) integrin to vitronectin. To investigate the role of this integrin in FGF-2-mediated cellular responses, we used immortalized murine brain capillary endothelial cells, denoted IBE cells. Because IBE cells proliferate and migrate in response to FGF-2-treatment, when cultured on fibronectin-coated surface, we first examined the inhibitory activity of this peptide on the binding of alpha(v)beta(3) integrin to fibronectin as well as vitronectin. Solid phase binding assay revealed that cRGDfV peptide strongly inhibited the binding of purified alpha(v)beta(3) integrin to vitonectin- and fibronectin-coated plastic surfaces at a concentration of 50 microM. cRGDfV peptide at 50 microM inhibited spreading as well as adhesion of IBE cells on vitronectin-coated plastic surface but not on fibronectin. On fibronectin-coated substrata, cRGDfV at 50 microM attenuated FGF-2-mediated chemotaxis, but not FGF-2-induced proliferation, of IBE cells. We have previously demonstrated that mitogen-activated protein kinase (MAPK) activation within focal adhesions through c-Src activity was involved in FGF-2-induced chemotaxis of IBE cells. Treatment of cells with cRGDfV peptide was associated with reduced c-Src activity without tyrosine dephosphorylation. Immunofluorescent staining showed that cRGDfV inhibited redistribution of c-Src into focal adhesions. MAPK activation by FGF-2 within focal adhesions was also attenuated in the presence of cRGDfV peptide. Our results indicated that cRGDfV peptide inhibited redistribution of c-Src into focal adhesions, leading to impaired MAPK activation within focal adhesions and motility in FGF-2-treated endothelial cells. PMID- 11478844 TI - Involvement of PKC-beta in PTH, TNF-alpha, and IL-1 beta effects on IL-6 promoter in osteoblastic cells and on PTH-stimulated bone resorption. AB - Protein kinase C (PKC) has been shown to be activated by parathyroid hormone (PTH) in osteoblasts. Prior evidence suggests that this activation mediates responses leading to bone resorption, including production of the osteoclastogenic cytokine interleukin-6 (IL-6). However, the importance of specific PKC isozymes in this process has not been investigated. A selective antagonist of PKC-beta, LY379196, was used to determine the role of the PKC-beta isozyme in the expression of IL-6 in UMR-106 rat osteoblastic cells and in bone resorption in fetal rat limb bone organ cultures. PTH, tumor necrosis factor alpha (TNF-alpha), and interleukin-1 beta (IL-1 beta) induced translocation of PKC-alpha and -beta(I) to the plasma membrane in UMR-106 cells within 5 min. The stimulation of PKC-beta(I) translocation by PTH, TNF-alpha or IL-1 beta was inhibited by LY379196. In contrast, LY379196 did not affect PTH, TNF-alpha-, or IL-1 beta-stimulated translocation of PKC-alpha. PTH, TNF-alpha, and IL-1 beta increased luciferase expression in UMR-106 cells transiently transfected with a 224/+11 bp IL-6 promoter-driven reporter construct. The IL-6 responses were also attenuated by treatment with LY379196. Furthermore, LY379196 inhibited bone resorption elicited by PTH in fetal rat bone organ cultures. These results indicate that PKC-beta(I) is a component of the signaling pathway that mediates PTH-, TNF-alpha-, and IL-1 beta-stimulated IL-6 expression and PTH-stimulated bone resorption. PMID- 11478845 TI - Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. AB - Transforming growth factor (TGF)-beta-induced chondrogenesis of mesenchymal stem cells derived from bone marrow involves the rapid deposition of a cartilage specific extracellular matrix. The sequential events in this pathway leading from the undifferentiated stem cell to a mature chondrocyte were investigated by analysis of key matrix elements. Differentiation was rapidly induced in cells cultured in the presence of TGF-beta 3 or -beta 2 and was accompanied by the early expression of fibromodulin and cartilage oligomeric matrix protein. An increase in aggrecan and versican core protein synthesis defined an intermediate stage, which also involved the small leucine-rich proteoglycans decorin and biglycan. This was followed by the appearance of type II collagen and chondroadherin. The pathway was also characterized by the appearance of type X collagen, usually associated with hypertrophic cartilage. There was also a change in the pattern of sulfation of chondroitin sulfate, with a progressive increase in the proportion of 6-sulfated species. The major proportion of newly synthesized glycosaminoglycan was part of an aggregating proteoglycan network. These data allow us to define the phenotype of the differentiated cell and to understand in greater detail the sequential process of matrix assembly. PMID- 11478846 TI - Replication timing of amplified genetic regions relates to intranuclear localization but not to genetic activity or G/R band. AB - Amplified genes in many human cancer cells usually localize at the extrachromosomal double minutes (DMs). In the present study, we show that multiple DMs in the human colorectal tumor COLO 320DM line replicated semisynchronously during the early S phase. On the other hand, during longer passage of the cells with DMs, cells with the amplified genes at the chromosomal homogeneously staining region (HSR) generally dominate the population. We currently report that HSR was composed of a tandem array of DM-derived sequences, which was shown using a unique DM-painting probe. Nevertheless, we found that HSR was replicated much later during the S phase, unless the amplified c-myc genes were expressed almost equally from DMs and HSR. Therefore, this provided a novel instance in which the cytogenetic localization affected replication timing without alteration of expression. Furthermore, we unexpectedly found that HSR had a distinctive band structure with respect to replication timing. The replication band structure was usually associated with the chromosomal G/R bands; however, HSR was homogeneous in the G/R band and in the distribution of highly repetitive sequences. We discuss the mechanism by which the replication band may arise, in relation to the folding of chromatin inside the nucleus. PMID- 11478847 TI - Identification and characterization of a novel fibronectin in zebrafish. AB - Sequence analysis of zebrafish fibronectin (FN) cDNAs indicates that at least two forms of the protein exist in fish. One form (FN1) is very similar to FNs identified in other vertebrates possessing 12 type I, 2 type II, and 17 type III repeats including two alternative splice sites (EIIIA and EIIIB) and a variable region (V). Zebrafish FN1 contains the RGD cell adhesion site in type III(10) and a second cell-binding site (LDV) in the V region. In addition to this conserved form of FN, a novel truncated form of zebrafish FN (FN2) was identified. The predicted structure of FN2 is identical to FN1 at the N-terminal region possessing 9 type I, 2 type II, and the first 3 type III repeats. Following III(3), FN2 contains a unique 20-amino-acid C-terminal tail that is different from the C-terminus of FN1, lacking the two cysteines that are usually involved in the formation of interchain disulfide bonds. Genomic sequence analysis has revealed that FN2 is generated by an alternative RNA splicing pattern that has not been described for FN in other organisms. Reverse transcription-polymerase chain reaction analysis and RNase protection assays reveal that FN2 mRNA is present in the zebrafish embryo throughout development as well as in cultures of an established liver cell line. Experiments conducted with recombinant FN2 synthesized in insect cells demonstrate that the protein promotes the attachment and spreading of fish embryo cells in culture. PMID- 11478848 TI - A key role for Pax7 transcripts in determination of muscle and nerve cells. AB - Embryonic teratocarcinoma mouse cells (P19) and embryonic NIH3T3 fibroblasts were induced chemically to differentiate along neurogenic or myogenic lineages. The expression profiles of Pax7 alternate transcripts were then assessed by RNA isolation and RT-PCR. Only two transcripts, Pax7b and Pax7d, were expressed in the neurogenic lineage. By contrast, in adult skeletal muscle, four transcripts, Pax7a-d, were expressed in the myogenic lineage. Moreover, P19 cells were shown to undergo neural cell differentiation when stably transfected with a single Pax7 transcript, PAX7b, generated from human skeletal muscle. Our results suggest a key role for Pax7 transcripts in lineage determination. PMID- 11478849 TI - Functional analysis of B144/LST1: a gene in the tumor necrosis factor cluster that induces formation of long filopodia in eukaryotic cells. AB - B144/LST1 is a gene encoded in the human major histocompatibility complex that produces multiple forms of alternatively spliced mRNA and encodes peptides fewer than 100 amino acids in length. B144/LST1 is strongly expressed in dendritic cells. Transfection of B144/LST1 into a variety of cells induces morphologic changes including the production of long, thin filopodia differing from those seen on transfection of a dominant active CDC42 gene. The structures are dynamically rearranging and sometimes connect one cell with another. The full effect of B144/LST1 protein on cell morphology requires the retention of at least one of the four cysteines of the peptide plus the presence of a hydrophobic segment in the protein, but requires only one of the two coding regions present in the terminal 3' exons. PMID- 11478850 TI - Centrosomal association of histone macroH2A1.2 in embryonic stem cells and somatic cells. AB - The histone 2A variant macroH2A1.2 is expressed in female and male mammals and is implicated in X-chromosome inactivation and autosomal gene silencing. In undifferentiated and early differentiating murine embryonic stem (ES) cells a cytosolic pool of macroH2A1.2 has recently been reported and found to be associated with the centrosome. Here, we show that the centrosomal association of macroH2A1.2 is a widespread phenomenon and is not restricted to undifferentiated and early differentiating ES cells. By indirect immunofluorescence we detect macroH2A1.2 protein in a juxtanuclear structure that duplicates once per cell cycle and colocalizes with centrosomal gamma-tubulin in both XX and XY ES cells prior to and throughout their differentiation. MacroH2A1.2 localization to the centrosome is also observed in female and male somatic cells, both in interphase and in mitosis. Biochemical analysis demonstrates that the association between macroH2A1.2 and the centrosome in somatic cells is stable, as macroH2A1.2 copurifies with centrosomes isolated from human lymphoblasts. Therefore, in addition to a nuclear pool of macroH2A1.2 a fraction of the histone is associated with the centrosome in various cell types and throughout ES cell differentiation. PMID- 11478851 TI - Characterization of human myocardial fibroblasts immortalized by HPV16 E6--E7 genes. AB - Human myocardial fibroblasts (HMF) have proved to be useful as a species specific cell culture system in various studies on myocarditis and cardiac remodelling. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary HMF with the E6 and E7 genes of the oncogenic human papillomavirus (HPV) 16. Successful transfection was demonstrated in 3 of 12 experiments by detection of E6-E7 gene transcription with nucleic acid sequence based amplification (NASBA). No significant change of phenotype was noted in the emerging cell lines (HMF(1226D), HMF(1321D), HMF(1226K)), but their in vitro lifespan was increased by 20 to 30 population doublings until cells entered crisis. A single subclone of HMF(1226K) had a transformed phenotype and continued to proliferate indefinitely. This subclone (HMF(1226K/I)) was considered to be immortalized and telomerase activity was detected. Despite the increased risk of mutations due to abrogation of p53 function, HMF(1226K/I) and the HMF lines with an increased lifespan retained the properties of primary HMF cells, as they expressed fibroblast markers (prolyl-4-hydroxylase, vimentin), cytokines (interleukin 1 alpha, 6, 8), and angiotensin II receptors and still were permissive for coxsackievirus B3 infection. PMID- 11478852 TI - Stimulation of mitogenic pathways through kinase-impaired mutants of the epidermal growth factor receptor. AB - Two residues have been shown to be critical for the kinase activity of the receptor for epidermal growth factor (EGF): lysine-721, which functions in the binding of ATP by correctly positioning the gamma-phosphate for phosphoryl transfer, and aspartate-813, which functions as the catalytic base of the kinase. Mutation of either of these two residues has been shown to disrupt kinase activity of the receptor. However, studies performed in different laboratories had suggested that while EGF receptors mutated at lysine-721 are unable to stimulate significant increases of [(3)H]thymidine incorporation into DNA in response to EGF treatment, cells expressing EGF receptors mutated at aspartate 813 do stimulate significant incorporation of [(3)H]thymidine into DNA in response to EGF. In the present study, EGF receptors mutated at lysine-721 or aspartate-813 (K721R and D813A, respectively), as well as wild-type EGF receptors, were expressed in the same cellular background, Chinese hamster ovary cells, and side-by-side experiments were performed to investigate possible signaling-related differences. Our results indicate that while there are measurable differences in the abilities of the two mutant receptors to stimulate [(3)H]thymidine incorporation between 20 and 24 h after addition of EGF, these differences cannot be correlated with significant differences in EGF-stimulated tyrosine phosphorylation of mutant EGF receptor and endogenous ErbB2, the extent of receptor internalization, EGF-stimulated ion uptake, stimulation of SHC activity, or receptor association with Grb2. Flow cytometric data suggest that populations of cells expressing either kinase-impaired mutant EGF receptor progress similarly into S phase in response to addition of EGF. These observations suggest that D813A and K721R retain similar ability to stimulate mitogenic signaling events through transactivation of ErbB2 with only subtle temporal differences, and they emphasize the importance of expressing mutant receptors in an identical cellular context to make valid comparisons of functions. PMID- 11478853 TI - Requirement for tyrosine kinase-ERK1/2 signaling in alpha 1 beta 1 integrin mediated collagen matrix remodeling by rat mesangial cells. AB - Abnormal mesangial extracellular matrix remodeling by mesangial cells (MCs) is the hallmark of progressive glomerulonephritis (GN). We recently showed, using a type I collagen gel contraction assay, that alpha 1 beta 1 integrin-dependent MC adhesion and migration are necessary cell behaviors for collagen matrix remodeling. To further determine the mechanism of alpha 1 beta 1 integrin mediated collagen remodeling, we studied the signaling pathways of MCs that participate in the regulation of collagen gel contraction. Immunoprecipitation and phosphotyrosine detection revealed that gel contraction is associated with the enhanced activity and phosphorylation of ERK1/2 by MCs. The tyrosine kinase inhibitors herbimycin and genistein inhibited collagen gel contraction dose dependently. Furthermore, targeting ERK1/2 activity with a MEK inhibitor, PD98059, and antisense ERK1/2 hindered gel contraction in a dose-dependent manner. Similar inhibitory effects on gel contraction and ERK1/2 phosphorylation were observed when MC-mediated gel contraction was performed in the presence of function-blocking anti-alpha1 or anti-beta1 integrin antibodies. However, cell adhesion and migration assays indicated that PD98059 and antisense ERK1/2 blocked alpha 1 beta 1 integrin-dependent MC migration, but did not interfere with collagen adhesion, although there was a marked decrease in ERK1/2 phosphorylation and ERK1/2 protein expression in cell adhesion on type I collagen. None of the above could affect membrane expression of alpha 1 beta 1 integrin. These results suggested that ERK1/2 activation is critical for the alpha 1 beta 1 integrin dependent MC migration necessary for collagen matrix reorganization. We therefore conclude that ERK1/2 may serve as a possible target for pharmacological inhibition of pathological collagen matrix formation in GN. PMID- 11478854 TI - Functional knockout of the corepressor CtBP by the second exon of adenovirus E1a relieves repression of transcription. AB - The C-terminal binding protein (CtBP) acts as a transcriptional corepressor upon recruitment to transcriptional regulators. In contrast, interaction between CtBP and the adenovirus E1A protein is required for efficient activation of E1A responsive genes, suggesting that E1A might block CtBP-mediated repression. Recruitment of CtBP to a promoter, either as a Gal4CtBP fusion or through an interaction with a Gal4 fusion protein expressing the CtBP interacting domain (CID) of E1A, resulted in transcriptional repression. The second exon of E1A, containing the CID, alleviated repression by Gal4E1ACID-recruited CtBP, but not Gal4CtBP-mediated repression, suggesting that E1A prevented repression by blocking promoter recruitment of CtBP. E1ACID was also sufficient to derepress transcription from several cotransfected promoter constructs. Furthermore, inducible expression of E1ACID in established cell lines resulted in significant changes of endogenous gene expression, possibly by sequestration of CtBP. Together, these data indicated that CtBP might act as a wide-range regulator of transcription. Although CtBP was shown to interact with histone deacetylases (HDACs), transcriptional repression by a Gal4CtBP fusion protein was not sensitive to inhibition of HDACs by trichostatin A (TSA). In contrast, TSA eliminated E1ACID derepression of E1A second exon-responsive promoters. Although the reason for this difference remains to be experimentally verified, it is possible that the requirement for HDACs might differ depending on the mechanism by which CtBP becomes promoter recruited. PMID- 11478855 TI - Phosphorylation of raf-1 by kinase suppressor of ras is inhibited by "MEK specific" inhibitors PD 098059 and U0126 in differentiating HL60 cells. AB - Determination of the involvement of MAP kinase cascades in signaling cell growth or differentiation is aided by the use of the inhibitors PD 098059 [2-(2'-amino 3'-methoxyphenyl)oxananphthalen-4-one] and U0126 [1,4-diamino-2,3-dicyano-1,4 bis(2-aminophenylthio)butadiene], believed to be MEK-specific kinase inhibitors. We report here that the activity of kinase suppressor of ras (KSR-1), a kinase upstream of raf-1, is inhibited by both these compounds at concentrations similar to those that inhibit MEK-1. Further, in HL60 cells induced to differentiate with 1,25-dihydroxyvitamin D(3) raf-1 and p90RSK, but not ERK1/2, are coregulated, and their expression as well as monocytic differentiation is inhibited in parallel by PD 098059. Thus, in this system raf-1 is phosphorylated by KSR-1, and PD 098059 as well as U0126 inhibits this phosphorylation. This suggests great caution in the interpretation of experiments that utilize these pharmacological inhibitors of kinase activity as evidence for a role for the MEK--ERK module in ras or raf-1 signaling. PMID- 11478856 TI - Bacteriophage lambda DNA packaging: DNA site requirements for termination and processivity. AB - Bacteriophage lambda chromosomes are processively packaged into preformed shells, using end-to-end multimers of intracellular viral DNA as the packaging substate. A 200 bp long DNA segment, cos, contains all the sequences needed for DNA packaging. The work reported here shows that efficient DNA packaging termination requires cos's I2 segment, in addition to the required termination subsite, cosQ, and the nicking site, cosN. Efficient processivity requires cosB, in addition to cosQ and cosN. An initiation-defective mutant form of cosB sponsored efficient processivity, indicating that the terminase-cosB interactions required for termination are less stringent than those required at initiation. The finding that an initiation-defective form of cosB is functional for processivity allows a re-interpretation of a similar finding, obtained previously, that the initiation defective cosB of phage 21 is functional for processivity by the lambda packaging machinery. The cosBphi21 result can now be interpreted as indicating that interactions between cosBphi21 and lambda terminase, while insufficient for initiation, function for processivity. PMID- 11478857 TI - Structural diversity of ex vivo amyloid fibrils studied by cryo-electron microscopy. AB - Cryo-electron microscopy studies are presented on amyloid fibrils isolated from amyloidotic organs of two patients with different forms of hereditary non neuropathic systemic amyloidosis, caused, respectively, by Leu60Arg apolipoprotein AI and Asp67His lysozyme. Although ex vivo amyloid fibrils were thought to be more uniform in structure than those assembled in vitro, our findings show that these fibrils are also quite variable in structure. Structural disorder and variability of the fibrils have precluded three-dimensional reconstruction, but averaged cryo-electron microscopy images suggest models for protofilament packing in the lysozyme fibrils. We conclude that ex vivo amyloid fibrils, although variable, assemble as characteristic structures according to the identity of the precursor protein. PMID- 11478858 TI - SIV gp41 binds to membranes both in the monomeric and trimeric states: consequences for the neuropathology and inhibition of HIV infection. AB - The viral envelope glycoprotein gp41 mediates membrane fusion in HIV/SIV infection. gp41 ectodomain (e-gp41, residues 27-149), which was shown to interact with phospholipid membranes, exists in an equilibrium between the monomeric and trimeric states. Here, we analyzed, by intrinsic Trp fluorescence and resonance energy transfer, whether SIV e-gp41-membrane interaction depends on the gp41 oligomeric state. We found that both gp41 monomers and trimers bind membranes, with the monomers' full binding being reached at substantially lower lipid to protein ratios. Furthermore, the different characteristics of the Trp fluorescence of monomers and trimers enabled us to detect binding of each form at concentrations at which both species were present. CD spectroscopy revealed that the secondary structure of gp41 monomers does not change upon membrane binding, suggesting that membrane-bound monomeric-gp41 is a possible target for DP-178, a potent peptide inhibitor of HIV infection. The consequences of the interaction between monomeric and trimeric gp41 with membranes in HIV/SIV infection, its inhibition, and its associated neuropathologies are discussed. PMID- 11478859 TI - Solution structure and interaction surface of the C-terminal domain from p47: a major p97-cofactor involved in SNARE disassembly. AB - p47 is the major protein identified in complex with the cytosolic AAA ATPase p97. It functions as an essential cofactor of p97-regulated membrane fusion, which has been suggested to disassemble t-t-SNARE complexes and prepare them for further rounds of membrane fusion. Here, we report the high-resolution NMR structure of the C-terminal domain from p47. It comprises a UBX domain and a 13 residue long structured N-terminal extension. The UBX domain adopts a characteristic ubiquitin fold with a betabetaalphabetabetaalphabeta secondary structure arrangement. Three hydrophobic residues from the N-terminal extension pack closely against a cleft in the UBX domain. We also identify, for the first time, the p97 interaction surface using NMR chemical shift perturbation studies. PMID- 11478860 TI - Roles of RNA:DNA hybrid stability, RNA structure, and active site conformation in pausing by human RNA polymerase II. AB - Human RNA polymerase II recognizes a strong transcriptional pause signal in the initially transcribed region of HIV-1. We report the use of a limited-step transcription assay to dissect the mechanism underlying recognition of and escape from this HIV-1 pause. Our results suggest that the primary determinant of transcriptional pausing is a relatively weak RNA:DNA hybrid that triggers backtracking of RNA polymerase II along the RNA and DNA chains and displaces the RNA 3' OH from the active site. In contrast, two alternative RNA secondary structures, TAR and anti-TAR, are not required for pausing and affect it only indirectly, rather than through direct interaction with RNA polymerase II. TAR accelerates escape from the pause, but anti-TAR inhibits formation of TAR prior to pause escape. The behavior of RNA polymerase II at a mutant pause signal supports a two-step, non-equilibrium mechanism in which the rate-determining step is a conformational change in the enzyme, rather than the changes in nucleic-acid base-pairing that accompany backtracking. PMID- 11478861 TI - Modularity and homology: modelling of the titin type I modules and their interfaces. AB - Titin is a giant muscle protein with a highly modular architecture consisting of multiple repeats of two sequence motifs, named type I and type II. Type I motifs are homologous to members of the fibronectin type 3 (Fn3) superfamily, one of the motifs most widespread in modular proteins. Fn3 domains are thought to mediate protein-protein interactions and to act as spacers. In titin, Fn3 modules are present in two different super-repeated patterns, likely to be involved in sarcomere assembly through interactions with A-band proteins. Here, we discuss results from homology modelling the whole family of Fn3 domains in titin. Homology modelling is a powerful tool that will play an increasingly important role in the post-genomic era. It is particularly useful for extending experimental structure determinations of parts of multidomain proteins that contain multiple copies of the same motif. The 3D structures of a representative titin type I domain and of other extracellular Fn3 modules were used as a template to model the structures of the 132 copies in titin. The resulting models suggest residues that contribute to the fold stability and allow us to distinguish these from residues likely to have functional importance. In particular, analysis of the models and mapping of the consensus sequence onto the 3D structure suggest putative surfaces of interaction with other proteins. From the structures of isolated modules and the pattern of conservation in the multiple alignment of the whole titin Ig and Fn3 families, it is possible to address the question of how tandem modules are assembled. Our predictions can be validated experimentally. PMID- 11478862 TI - Structure and mechanism of the RuvB Holliday junction branch migration motor. AB - The RuvB hexamer is the chemomechanical motor of the RuvAB complex that migrates Holliday junction branch-points in DNA recombination and the rescue of stalled DNA replication forks. The 1.6 A crystal structure of Thermotoga maritima RuvB together with five mutant structures reveal that RuvB is an ATPase-associated with diverse cellular activities (AAA+-class ATPase) with a winged-helix DNA binding domain. The RuvB-ADP complex structure and mutagenesis suggest how AAA+ class ATPases couple nucleotide binding and hydrolysis to interdomain conformational changes and asymmetry within the RuvB hexamer implied by the crystallographic packing and small-angle X-ray scattering in solution. ATP-driven domain motion is positioned to move double-stranded DNA through the hexamer and drive conformational changes between subunits by altering the complementary hydrophilic protein- protein interfaces. Structural and biochemical analysis of five motifs in the protein suggest that ATP binding is a strained conformation recognized both by sensors and the Walker motifs and that intersubunit activation occurs by an arginine finger motif reminiscent of the GTPase-activating proteins. Taken together, these results provide insights into how RuvB functions as a motor for branch migration of Holliday junctions. PMID- 11478863 TI - Detailed analysis of RNA-protein interactions within the ribosomal protein S8 rRNA complex from the archaeon Methanococcus jannaschii. AB - The crystal structure of ribosomal protein S8 bound to its target 16 S rRNA from a hyperthermophilic archaeon Methanococcus jannaschii has been determined at 2.6 A resolution. The protein interacts with the minor groove of helix H21 at two sites located one helical turn apart, with S8 forming a bridge over the RNA major groove. The specificity of binding is essentially provided by the C-terminal domain of S8 and the highly conserved nucleotide core, characterized by two dinucleotide platforms, facing each other. The first platform (A595-A596), which is the less phylogenetically and structurally constrained, does not directly contact the protein but has an important shaping role in inducing cross-strand stacking interactions. The second platform (U641-A642) is specifically recognized by the protein. The universally conserved A642 plays a pivotal role by ensuring the cohesion of the complex organization of the core through an array of hydrogen bonds, including the G597-C643-U641 base triple. In addition, A642 provides the unique base-specific interaction with the conserved Ser105, while the Thr106 - Thr107 peptide link is stacked on its purine ring. Noteworthy, the specific recognition of this tripeptide (Thr-Ser-Thr/Ser) is parallel to the recognition of an RNA tetraloop by a dinucleotide platform in the P4-P6 ribozyme domain of group I intron. This suggests a general dual role of dinucleotide platforms in recognition of RNA or peptide motifs. One prominent feature is that conserved side-chain amino acids, as well as conserved bases, are essentially involved in maintaining tertiary folds. The specificity of binding is mainly driven by shape complementarity, which is increased by the hydrophobic part of side-chains. The remarkable similarity of this complex with its homologue in the T. thermophilus 30 S subunit indicates a conserved interaction mode between Archaea and Bacteria. PMID- 11478864 TI - Dependence on solution conditions of aggregation and amyloid formation by an SH3 domain. AB - The formation of amyloid fibrils by the SH3 domain of the alpha-subunit of bovine phosphatidylinositol-3'-kinase (PI3-SH3) has been investigated under carefully controlled solution conditions. NMR and CD characterisation of the denatured states from which fibrils form at low pH show that their properties can be correlated with the nature of the resulting aggregates defined by EM and FTIR spectroscopy. Compact partially folded states, favoured by the addition of anions, are prone to precipitate rapidly into amorphous species, whilst well defined fibrillar structures are formed slowly from more expanded denatured states. Kinetic data obtained by a variety of techniques show a clear lag phase in the formation of amyloid fibrils. NMR spectroscopy shows no evidence for a significant population of small oligomers in solution during or after this lag phase. EM and FTIR indicate the presence of amorphous aggregates (protofibrils) rich in beta-structure after the lag phase but prior to the development of well defined amyloid fibrils. These observations strongly suggest a nucleation and growth mechanism for the formation of the ordered aggregates. The morphologies of the fibrillar structures were found to be highly sensitive to the pH at which the protein solutions are incubated. This can be attributed to the effect of small perturbations in the electrostatic interactions that stabilise the contacts between the protofilaments forming the amyloid fibrils. Moreover, different hydrogen bonding patterns related to the various aggregate morphologies can be distinguished by FTIR analysis. PMID- 11478865 TI - NMR structure and backbone dynamics of a concatemer of epidermal growth factor homology modules of the human low-density lipoprotein receptor. AB - The ligand-binding region of the low-density lipoprotein (LDL) receptor is formed by seven N-terminal, imperfect, cysteine-rich (LB) modules. This segment is followed by an epidermal growth factor precursor homology domain with two N terminal, tandem, EGF-like modules that are thought to participate in LDL binding and recycling of the endocytosed receptor to the cell surface. EGF-A and the concatemer, EGF-AB, of these modules were expressed in Escherichia coli. Correct protein folding of EGF-A and the concatemer EGF-AB was achieved in the presence or absence of calcium ions, in contrast to the LB modules, which require them for correct folding. Homonuclear and heteronuclear 1H-15N NMR spectroscopy at 17.6 T was used to determine the three-dimensional structure of the concatemer. Both modules are formed by two pairs of short, anti-parallel beta-strands. In the concatemer, these modules have a fixed relative orientation, stabilized by calcium ion-binding and hydrophobic interactions at the interface. 15N longitudinal and transverse relaxation rates, and [1H]-15N heteronuclear NOEs were used to derive a model-free description of the backbone dynamics of the molecule. The concatemer appears relatively rigid, particularly near the calcium ion-binding site at the module interface, with an average generalized order parameter of 0.85+/-0.11. Some mutations causing familial hypercholesterolemia may now be rationalized. Mutations of D41, D43 and E44 in the EGF-B calcium ion binding region may affect the stability of the linker and thus the orientation of the tandem modules. The diminutive core also provides little structural stabilization, necessitating the presence of disulfide bonds. The structure and dynamics of EGF-AB contrast with the N-terminal LB modules, which require calcium ions both for folding to form the correct disulfide connectivities and for maintenance of the folded structure, and are connected by highly mobile linking peptides. PMID- 11478866 TI - Bergerac-SH3: "frustation" induced by stabilizing the folding nucleus. AB - The influence of an inserted exogenous independent folding element on the thermodynamics and folding properties of SH3 domain from alpha-spectrin has been investigated by creating a fused form between this small all-beta domain and a stable beta-hairpin (BH19). NMR analysis of synthetic peptides shows that insertion of BH19 nucleates formation of the original natural beta-hairpin (distal loop) that is part of the SH3 folding nucleus. The resulting protein (Bergerac-SHH) is more stable, folds faster and contains an elongated hairpin protruding from the globular domain as determined by 2D-NMR. "Protein engineering" analysis of the inserted region shows that it is folded in the transition state. Interestingly, stabilisation by insertion of the distal loop region results in the appearance of a compact intermediate revealed by a curved chevron plot at low denaturant concentration. This effect is eliminated at low salt concentrations by a single mutation of a hydrophobic residue within BH19 sequence, which is most probably involved in non-native interactions. Local stabilisation by enlargement and reinforcement of the folding nucleus, global compaction by the addition of salt and non-native interactions are shown to contribute to the observed deviation from the two-state behaviour. PMID- 11478867 TI - The folding mechanism of a beta-sheet: the WW domain. AB - The folding thermodynamics and kinetics of the Pin WW domain, a three-stranded antiparallel beta-sheet, have been characterized extensively. Folding and activation free energies were determined as a function of temperature for 16 mutants, which sample all strands and turns of the molecule. The mutational phi value (Phi(m)) diagram is a smooth function of sequence, indicating a prevalence of local interactions in the transition state (TS). At 37 degrees C, the diagram has a single pronounced maximum at turn 1: the rate-limiting step during folding is the formation of loop 1. In contrast, key residues for thermodynamic stability are located in the strand hydrophobic clusters, indicating that factors contributing to protein stability and folding kinetics are not correlated. The location of the TS along the entropic reaction coordinate Phi(T), obtained by temperature-tuning the kinetics, reveals that sufficiently destabilizing mutants in loop 2 or in the Leu7-Trp11-Tyr24-Pro37 hydrophobic cluster can cause a switch to a late TS. Phi(m) analysis is usually applied "perturbatively" (methyl truncation), but with Phi(T) to quantitatively assess TS shifts along a reaction coordinate, more severe mutations can be used to probe regions of the free energy surface beyond the TS. PMID- 11478868 TI - Automated structure-based prediction of functional sites in proteins: applications to assessing the validity of inheriting protein function from homology in genome annotation and to protein docking. AB - A major problem in genome annotation is whether it is valid to transfer the function from a characterised protein to a homologue of unknown activity. Here, we show that one can employ a strategy that uses a structure-based prediction of protein functional sites to assess the reliability of functional inheritance. We have automated and benchmarked a method based on the evolutionary trace approach. Using a multiple sequence alignment, we identified invariant polar residues, which were then mapped onto the protein structure. Spatial clusters of these invariant residues formed the predicted functional site. For 68 of 86 proteins examined, the method yielded information about the observed functional site. This algorithm for functional site prediction was then used to assess the validity of transferring the function between homologues. This procedure was tested on 18 pairs of homologous proteins with unrelated function and 70 pairs of proteins with related function, and was shown to be 94 % accurate. This automated method could be linked to schemes for genome annotation. Finally, we examined the use of functional site prediction in protein-protein and protein-DNA docking. The use of predicted functional sites was shown to filter putative docked complexes with a discrimination similar to that obtained by manually including biological information about active sites or DNA-binding residues. PMID- 11478869 TI - Dynamics of hydration in hen egg white lysozyme. AB - We investigate the hydration dynamics of a small globular protein, hen egg-white lysozyme. Extensive simulations (two trajectories of 9 ns each) were carried out to identify the time-scales and mechanism of water attachment to this protein. The location of the surface and integral water molecules in lysozyme was also investigated. Three peculiar temporal scales of the hydration dynamics can be discerned: two among these, with sub-nanosecond mean residence time, tau(w), are characteristic of surface hydration water; the slower time-scale (tau(w) approximately 2/3 ns) is associated with buried water molecules in hydrophilic pores and in superficial clefts. The computed tau(w) values in the two independent runs fall in a similar range and are consistent with each other, thus adding extra weight to our result. The tau(w) of surface water obtained from the two independent trajectories is 20 and 24 ps. In both simulations only three water molecules are bound to lysozyme for the entire length of the trajectories, in agreement with nuclear magnetic relaxation dispersion estimates. Locations other than those identified in the protein crystal are found to be possible for these long-residing water molecules. The dynamics of the hydration water molecules observed in our simulations implies that each water molecule visits a multitude of residues during the lifetime of its bound with the protein. The number of residues seen by a single water molecule increases with the time-scale of its residence time and, on average, is equal to one only for the water molecules with shorter residence time. Thus, tau(w) values obtained from inelastic neutron scattering and based on jump-diffusion models are likely not to account for the contribution of water molecules with longer residence time. PMID- 11478870 TI - Extending the accuracy limits of prediction for side-chain conformations. AB - Current techniques for the prediction of side-chain conformations on a fixed backbone have an accuracy limit of about 1.0-1.5 A rmsd for core residues. We have carried out a detailed and systematic analysis of the factors that influence the prediction of side-chain conformation and, on this basis, have succeeded in extending the limits of side-chain prediction for core residues to about 0.7 A rmsd from native, and 94 % and 89 % of chi(1) and chi(1+2 ) dihedral angles correctly predicted to within 20 degrees of native, respectively. These results are obtained using a force-field that accounts for only van der Waals interactions and torsional potentials. Prediction accuracy is strongly dependent on the rotamer library used. That is, a complete and detailed rotamer library is essential. The greatest accuracy was obtained with an extensive rotamer library, containing over 7560 members, in which bond lengths and bond angles were taken from the database rather than simply assuming idealized values. Perhaps the most surprising finding is that the combinatorial problem normally associated with the prediction of the side-chain conformation does not appear to be important. This conclusion is based on the fact that the prediction of the conformation of a single side-chain with all others fixed in their native conformations is only slightly more accurate than the simultaneous prediction of all side-chain dihedral angles. PMID- 11478871 TI - Coupling of conformational folding and disulfide-bond reactions in oxidative folding of proteins. AB - The oxidative folding of proteins consists of conformational folding and disulfide-bond reactions. These two processes are coupled significantly in folding-coupled regeneration steps, in which a single chemical reaction (the "forward" reaction) converts a conformationally unstable precursor species into a conformationally stable, disulfide-protected successor species. Two limiting-case mechanisms for folding-coupled regeneration steps are described. In the folded precursor mechanism, the precursor species is preferentially folded at the moment of the forward reaction. The (transient) native structure increases the effective concentrations of the reactive thiol and disulfide groups, thus favoring the forward reaction. By contrast, in the quasi-stochastic mechanism, the forward reaction occurs quasi-stochastically in an unfolded precursor; i.e., reactive groups encounter each other with a probability determined primarily by loop entropy, albeit modified by conformational biases in the unfolded state. The resulting successor species is initially unfolded, and its folding competes with backward chemical reactions to the unfolded precursors. The folded-precursor and quasi-stochastic mechanisms may be distinguished experimentally by the dependence of their kinetics on factors affecting the rates of thiol--disulfide exchange and conformational (un)folding. Experimental data and structural and biochemical arguments suggest that the quasi-stochastic mechanism is more plausible than the folded-precursor mechanism for most proteins. PMID- 11478872 TI - Beta-barrel pore-forming toxins: intriguing dimorphic proteins. PMID- 11478873 TI - Mechanism of unfolding of a model helical peptide. AB - Synthetic model helical peptides, Acetyl-W(EAAAR)(5)A-amide with (13)C=O specifically labeled alanine segments in repeats n = 1,2 or 4,5 were studied in aqueous D(2)O solution as a function of temperature using Fourier transform infrared spectroscopy and two-dimensional correlation analysis. The (13)C==O provided a probe which was sensitive to the carbonyl stretch in the peptide bonds of the alanine residues at the amino terminal end in one peptide as compared to the probe in the carboxy terminal end of the other peptide during thermal perturbation. The relative stability of each terminal end was examined; the more stable terminal was determined to be the amino terminal end. Also studied were the glutamate and arginine side-chain modes involved in the salt bridging interaction. Two-dimensional correlation analysis enabled enhanced resolution in the spectral region of 1520--1700 cm(-1), and thus, the order in which these vibrational modes were perturbed as a function of increasing temperature were established. PMID- 11478874 TI - Solution structure of the satiety factor, CART, reveals new functionality of a well-known fold. AB - Cocaine and amphetamine regulated transcript (CART) peptide has been shown to be an anorectic peptide that inhibits both normal and starvation-induced feeding and completely blocks the feeding response induced by neuropeptide Y and regulated by leptin in the hypothalamus. The C-terminal part containing the three disulfide bridges CART(48-89) is the biologically active part of the molecule affecting food intake. The solution structure of the active part of CART has a fold equivalent to other functionally distinct small proteins. CART consists mainly of turns and loops spanned by a compact framework composed by a few small stretches of antiparallel beta-sheet common to cystine knots. PMID- 11478875 TI - Identification of a subunit interface in transthyretin amyloid fibrils: evidence for self-assembly from oligomeric building blocks. AB - Amyloid and prion diseases appear to stem from the conversion of normally folded proteins into insoluble, fiber-like assemblies. Despite numerous structural studies, a detailed molecular characterization of amyloid fibrils remains elusive. In particular, models of amyloid fibrils proposed thus far have not adequately defined the constituent protein subunit interactions. To further our understanding of amyloid structure, we employed thiol-specific cross-linking and site-directed spin labeling to identify specific protein-protein associations in transthyretin (TTR) amyloid fibrils. We find that certain cysteine mutants of TTR, when dimerized by chemical cross-linkers, still form fibers under typical in vitro fibrillogenic conditions. In addition, site-directed spin labeling of many residues at the natural dimer interface reveals that their spatial proximity is preserved in the fibrillar state even in the absence of cross-linking constraints. Here, we present the first view of a subunit interface in TTR fibers and show that it is very similar to one of the natural dimeric interchain associations evident in the structure of soluble TTR. The results clarify varied models of amyloidogenesis by demonstrating that transthyretin amyloid fibrils may assemble from oligomeric protein building blocks rather than structurally rearranged monomers. PMID- 11478876 TI - Contribution of chain termini to the conformational stability and biological activity of onconase. AB - Onconase, a member of the RNase A superfamily, is a potent antitumor agent which is undergoing phase III clinical trials as an antitumor drug. We have recently shown that onconase is an unusually stable protein. Furthermore, the protein is resistant to the action of proteases, which could influence its use as a drug, prolonging its biological life, and leading to its renal toxicity. Our investigation focused on the contribution of chain termini to onconase conformational stability and biological activities. We used differential scanning calorimetry, isothermal unfolding experiments, limited proteolysis, and catalytic and antitumor activity determinations to investigate the effect of the elimination of the two blocks at the chain termini, the N-terminal cyclized glutamine and the C-terminal disulfide bridge between the terminal Cys104 and Cys87. The determination of the thermodynamic parameters of the protein led to the conclusion that the two blocks at onconase chain termini are responsible for the unusual stability of the protein. Moreover, the reduced stability of the onconase mutants does not influence greatly their catalytic and antitumor activities. Thus, our data would suggest that an onconase-based drug, with a decreased toxicity, could be obtained through the use of less stable onconase variants. PMID- 11478877 TI - Active center cleft residues of pokeweed antiviral protein mediate its high affinity binding to the ribosomal protein L3. AB - Pokeweed antiviral protein (PAP) is a ribosome-inactivating protein (RIP) which catalytically cleaves a specific adenine base from the highly conserved alpha sarcin/ricin loop (SRL) of the large ribosomal RNA and thereby inhibits the protein synthesis. The ribosomal protein L3, a highly conserved protein located at the peptidyltransferase center of the ribosomes, is involved in binding of PAP to ribosomes and subsequent depurination of the SRL. We have recently discovered that recombinant PAP mutants with alanine substitution of the active center cleft residues (69)NN(70) (FLP-4) and (90)FND(92) (FLP-7) that are not directly involved in the catalytic depurination at the active site exhibit >150-fold reduced ribosome inhibitory activity [(2000) J. Biol. Chem. 275, 3382--3390]. We hypothesized that the partially exposed half of the active site cleft could be the potential docking site for the L3 molecule. Our modeling studies presented herein indicated that PAP residues 90--96, 69--70, and 118--120 potentially interact with L3. Therefore, mutations of these residues were predicted to result in destabilization of interactions with rRNA and lead to a lower binding affinity with L3. In the present structure-function relationship study, coimmunoprecipitation assays with an in vitro synthesized yeast ribosomal protein L3 suggested that these mutant PAP proteins poorly interact with L3. The binding affinities of the mutant PAP proteins for ribosomes and recombinant L3 protein were calculated from rate constants and analysis of binding using surface plasmon resonance biosensor technology. Here, we show that, compared to wild-type PAP, FLP-4/(69)AA(70) and FLP-7/(90)AAA(92) exhibit significantly impaired affinity for ribosomes and L3 protein, which may account for their inability to efficiently inactivate ribosomes. By comparison, recombinant PAP mutants with alanine substitutions of residues (28)KD(29) and (111)SR(112) that are distant from the active center cleft showed normal binding affinity to ribosomes and L3 protein. The single amino acid mutants of PAP with alanine substitution of the active center cleft residues N69 (FLP-20), F90 (FLP-21), N91 (FLP-22), or D92 (FLP-23) also showed reduced ribosome binding as well as reduced L3 binding, further confirming the importance of the active center cleft for the PAP- ribosome and PAP--L3 interactions. The experimental findings presented in this report provide unprecedented evidence that the active center cleft of PAP is important for its in vitro binding to ribosomes via the L3 protein. PMID- 11478878 TI - Mechanism of specific target recognition and RNA hydrolysis by ribonucleolytic toxin restrictocin. AB - Restrictocin, a member of the fungal ribotoxin family, specifically cleaves a single phosphodiester bond in the 28S rRNA and potently inhibits eukaryotic protein synthesis. Residues Tyr47, His49, Glu95, Phe96, Pro97, Arg120, and His136 have been predicted to form the active site of restrictocin. In this study, we have individually mutated these amino acids to alanine to probe their role in restrictocin structure and function. The role of Tyr47, His49, Arg120, and His136 was further investigated by making additional mutants. Mutating Arg120 or His136 to alanine or the other amino acids rendered the toxin completely inactive, whereas mutating Glu95 to alanine only partially inactivated the toxin. Mutation of Phe96 and Pro97 to Ala had no effect on the activity of restrictocin. The Tyr47 to alanine mutant was inactive in inhibiting protein synthesis, and had a nonspecific ribonuclease activity on 28S rRNA similar to that shown previously for the His49 to Ala mutant. Unlike the His136 to Ala mutant, the double mutants containing Tyr47 or His49 mutated to alanine along with His136 did not compete with restrictocin to cause a significant reduction in the extent of cleavage of 28S rRNA. In a model of restrictocin and a 29-mer RNA substrate complex, residues Tyr47, His49, Glu95, Arg120, and His136 were found to be near the cleavage site on RNA. It is proposed that in restrictocin Glu95 and His136 are directly involved in catalysis, Arg120 is involved in the stabilization of the enzyme substrate complex, Tyr47 provides structural stability to the active site, and His49 determines the substrate specificity. PMID- 11478879 TI - Species specificity of amidine-based urokinase inhibitors. AB - Inhibition of the proteolytic activity of urokinase has been shown to inhibit the progression of tumors in rodent models and is being investigated for use in human disease. Understanding the rodent/human species-specificity of urokinase inhibitors is therefore critical for interpretation of rodent cancer progression models that use these inhibitors. We report here studies with a panel of 11 diverse urokinase inhibitors in both human and mouse enzymatic assays. Inhibitors such as amiloride, B428, and naphthamidine, that occupy only the S1 subsite pocket were found to be nearly equipotent between the human and the murine enzymes. Inhibitors that access additional, more distal, pockets were significantly more potent against the human enzyme but there was no corresponding potency increase against the murine enzyme. X-ray crystallographic structures of these compounds bound to the serine protease domain of human urokinase were solved and examined in order to explain the human/mouse potency differences. The differences in inhibitor potency could be attributed to four amino acid residues that differ between murine and human urokinases: 60, 99, 146, and 192. These residues are Asp, His, Ser, and Gln in human and Gln, Tyr, Glu, and Lys in mouse, respectively. Compounds bearing a cationic group that interacts with residue 60 will preferentially bind to the human enzyme because of favorable electrostatic interactions. The hydrogen bonding to residue 192 and steric considerations with residues 99 and 146 also contribute to the species specificity. The nonparallel human/mouse enzyme inhibition observations were extended to a cell-culture assay of urokinase-activated plasminogen-mediated fibronectin degradation with analogous results. These studies will aid the interpretation of in vivo evaluation of urokinase inhibitors. PMID- 11478880 TI - Alternate substrate binding modes to two mutant (D98N and H255N) forms of nitrite reductase from Alcaligenes faecalis S-6: structural model of a transient catalytic intermediate. AB - High-resolution nitrite soaked oxidized and reduced crystal structures of two active site mutants, D98N and H255N, of nitrite reductase (NIR) from Alcaligenes faecalis S-6 were determined to better than 2.0 A resolution. In the oxidized D98N nitrite-soaked structures, nitrite is coordinated to the type II copper via its oxygen atoms in an asymmetric bidentate manner; however, elevated B-factors and weak electron density indicate that both nitrite and Asn98 are less ordered than in the native enzyme. This disorder likely results from the inability of the N delta 2 atom of Asn98 to form a hydrogen bond with the bound protonated nitrite, indicating that the hydrogen bond between Asp98 and nitrite in the native NIR structure is essential in anchoring nitrite in the active site for catalysis. In the oxidized nitrite soaked H255N crystal structure, nitrite does not displace the ligand water and is instead coordinated in an alternative mode via a single oxygen to the type II copper. His255 is clearly essential in defining the nitrite binding site despite the lack of direct interaction with the substrate in the native enzyme. The resulting pentacoordinate copper site in the H255N structure also serves as a model for a proposed transient intermediate in the catalytic mechanism consisting of a hydroxyl and nitric oxide molecule coordinated to the copper. The formation of an unusual dinuclear type I copper site in the reduced nitrite soaked D98N and H255N crystal structures may represent an evolutionary link between the mononuclear type I copper centers and dinuclear Cu(A) sites. PMID- 11478881 TI - Dynamics of the Mrf-2 DNA-binding domain free and in complex with DNA. AB - Mrf-2 is a member of a new class of DNA-binding proteins known as the AT-rich interaction domain family or ARID. Chemical shift indices and characteristic NOE values indicate that the three-dimensional structure of the Mrf-2 ARID in complex with DNA is nearly identical to that of the free protein. The backbone dynamics of the Mrf-2 domain free and in complex with DNA have been characterized by (15)N NMR relaxation measurements and model-free analysis. Chemical shift perturbations and dynamic studies suggest that two flexible interhelical loops, the flexible C terminal tail, and one alpha-helix are involved in DNA recognition, indicating the importance of protein dynamics in DNA binding. Some well-structured regions, in particular the putative DNA-contacting helix, in Mrf-2 show a decrease in the order parameters (S(2)) upon complex formation. The less well-structured loops and the unstructured C-terminus show reduced flexibility upon DNA binding. In addition, the model-free analysis indicates motions on the picosecond to nanosecond and micro- to millisecond time scales at the DNA-binding surface of the bound Mrf-2 ARID, suggesting a model where interactions between the protein and DNA are highly dynamic. PMID- 11478882 TI - Modifications of cysteine residues in the solution and membrane-associated conformations of phosphatidylinositol transfer protein have differential effects on lipid transfer activity. AB - The alpha isoforms of mammalian phosphatidylinositol transfer protein (PITP) contain four conserved Cys residues. In this investigation, a series of thiol modifying reagents, both alkylating and mixed disulfide-forming, was employed to define the accessibility of these residues and to evaluate their role in protein mediated intermembrane phospholipid transport. Isolation and analysis of chemically modified peptides and site-directed mutagenesis of each Cys residue to Ala were also performed. Soluble, membrane-associated, and denatured preparations of wild-type and mutant rat PITPs were studied. Under denaturing conditions, all four Cys residues could be detected spectrophotometrically by chemical reaction with 4,4'-dipyridyl disulfide or 5,5'-dithiobis(2-nitrobenzoate). In the native protein, two of the four Cys residues were sensitive to some but not all thiol modifying reagents, with discrimination based on the charge and hydrophobicity of the reagent and the conformation of the protein. With the soluble conformation of PITP, achieved in the absence of phospholipid vesicles, the surface-exposed Cys(188) was chemically modified without consequence to lipid transfer activity. Cys(188) exhibited an apparent pK(a) of 7.6. The buried Cys(95), which constitutes part of the phospholipid substrate binding site, was covalently modified upon transient association of PITP with a membrane surface. The Cys-to Ala mutations showed that neither Cys(95) nor Cys(188) was essential for lipid transfer activity. However, chemical modification of Cys(95) resulted in the loss of lipid transfer activity. These results demonstrate that the Cys residues of PITP can be assigned to several different classes of chemical reactivity. Of particular interest is Cys(95), whose sulfhydryl group becomes exposed to modification in the membrane-associated conformation of PITP. Furthermore, the inhibition of PITP activity by thiol-modifying reagents is a result of steric hindrance of phospholipid substrate binding. PMID- 11478883 TI - Recognition of fibronectin by the platelet integrin alpha IIb beta 3 involves an extended interface with multiple electrostatic interactions. AB - Normal platelet function is dependent on the ability of integrin alpha IIb beta 3 (glycoprotein IIb/IIIa) to interact with components of the subendothelial matrix, such as fibronectin (Fn), exposed at sites of vascular injury. Studies using synthetic peptides derived from human Fn sequences Asp(1373)--Thr(1383) and Arg(1493)--Asp(1495) have suggested a role for both the 9th (3fn9) and 10th (3fn10) type III repeats of this ligand in binding to alpha IIb beta 3. In this study, we have taken a charge-to-alanine mutagenesis approach to evaluate the importance of these sites, and other charged residues, within the context of recombinant 3fn9--10 modules for binding to alpha IIb beta 3. To identify residues that are involved in Fn binding to alpha IIb beta 3, recombinantly expressed 3fn9--10 module pairs with alanine substitutions introduced into each of the 38 charged residues were individually assayed for the ability to inhibit Fn binding to purified alpha IIb beta 3. Substitutions at Fn residues Arg(1493) and Asp(1495) of the RGD sequence were found to have the greatest effect on alpha IIb beta 3 binding, as expected. However, Fn residues Arg(1369), Arg(1371), Arg(1379), Arg(1445), and Arg(1448) were needed for optimal interaction of the 3fn9--10 module pair with alpha IIb beta 3. All Fn residues found to affect binding of 3fn9--10 to alpha IIb beta 3 are located on the same face and extend from the surface of the molecule. Additionally, the epitopes for two anti-Fn monoclonal antibodies that inhibit binding of this ligand to alpha IIb beta 3 were found to overlap the sites identified. These results demonstrate that alpha IIb beta 3--Fn binding involves multiple electrostatic interactions. PMID- 11478884 TI - Clostridium thermocellum Xyn10B carbohydrate-binding module 22-2: the role of conserved amino acids in ligand binding. AB - The majority of plant cell wall hydrolases are modular enzymes which, in addition to a catalytic module, possess one or more carbohydrate-binding modules (CBMs). These carbohydrate-active enzymes and their constituent modules have been classified into a number of families based upon amino acid sequence similarity. The Clostridium thermocellum xylanase, Xyn10B, contains two CBMs that belong to family 22 (CBM22). The crystal structure of the C-terminal CBM22 (CBM22-2) was determined in a previous study [Charnock, S. J., et al. (2000) Biochemistry 39, 5013--5021] and revealed a surface cleft which presents several conserved residues that are implicated in ligand binding. These amino acids have been substituted and the structure and biochemical properties of the mutants analyzed. The data show that R25A, W53A, Y103A, Y136A, and E138A exhibit greatly reduced affinity for xylotetraose relative to that of the wild-type protein. Conversely, mutations Y103F and Y136F have little effect on ligand binding. Using thermodynamic, X-ray, and NMR measurements on the mutants, we show that the cleft of CBM22-2 does indeed form the ligand-binding site. Trp 53 and Tyr 103 most likely participate in hydrophobic stacking interactions with the ligand, while Glu 138 makes one or more important hydrogen bonds with the tetrasaccharide. Although Arg 25 and Tyr 136 are likely to form hydrogen bonds with the ligand, they are also shown to play a critical role in maintaining the structural integrity of the binding cleft. PMID- 11478885 TI - Pre-steady-state kinetic studies of Saccharomyces cerevisiae myristoylCoA:protein N-myristoyltransferase mutants identify residues involved in catalysis. AB - MyristoylCoA:protein N-myristoyltransferase (Nmt, EC 2.3.1.97), a member of the GCN5 acetyltransferase (GNAT) superfamily, is an essential eukaryotic enzyme that catalyzes covalent attachment of myristate (C14:0) to the N-terminal Gly of proteins involved in myriad cellular functions. The 2.5 A resolution structure of a ternary complex of Saccharomyces cerevisiae Nmt1p with a bound substrate peptide (GLYASKLA) and nonhydrolyzable myristoylCoA analogue [Farazi, T. A., et al. (2001) Biochemistry 40, 6335] was used as the basis for a series of mutagenesis experiments designed to define the enzyme's catalytic mechanism. The kinetic properties of an F170A/L171A Nmt mutant are consistent with the proposal that their main chain amides, located in a beta-bulge structure conserved among GNATs, function as an oxyanion hole to polarize the thioester carbonyl of bound myristoylCoA prior to subsequent nucleophilic attack. Removal of the two C terminal residues (M454 and L455) produces a 300--400-fold reduction in the chemical transformation rate and converts the rate-limiting step from a step after the transformation to the transformation event itself. This finding is consistent with the main chain C-terminal carboxylate of L455 functioning as a catalytic base that abstracts a proton from the N-terminal Gly ammonium of the bound peptide to generate the nucleophilic amine. Mutating N169 and T205 in concert reduces the rate of the chemical transformation, supporting their role as components of an H-bonding network that facilitates attack of the Gly1 amine and stabilizes the tetrahedral intermediate. PMID- 11478886 TI - Mechanistic roles of Thr134, Tyr160, and Lys 164 in the reaction catalyzed by dTDP-glucose 4,6-dehydratase. AB - Escherichia coli dTDP-glucose 4,6-dehydratase and UDP-galactose 4-epimerase are members of the short-chain dehydrogenase/reductase SDR family. A highly conserved triad consisting of Ser/Thr, Tyr, and Lys is present in the active sites of these enzymes as well in other SDR proteins. Ser124, Tyr149, and Lys153 in the active site of UDP-galactose 4-epimerase are located in similar positions as the corresponding Thr134, Tyr160, and Lys164, in the active site of dTDP-glucose 4,6 dehydratase. The role of these residues in the first hydride transfer step of the dTDP-glucose 4,6-dehydratase mechanism has been studied by mutagenesis and steady state kinetic analysis. In all mutants except T134S, the k(cat) values are more than 2 orders of magnitude lower than of wild-type enzyme. The substrate analogue, dTDP-xylose, was used to investigate the effects of the mutations on rate of the first hydride transfer step. The first step becomes significantly rate limiting upon mutation of Tyr160 to Phe and only partly rate limiting in the reaction catalyzed by K164M and T134A dehydratases. The pH dependence of k(cat), the steady-state NADH level, and the fraction of NADH formed with saturating dTDP xylose show shifts in the pK(a) assigned to Tyr160 to more basic values by mutation of Lys164 and Thr134. The pK(a) of Tyr160, as determined by the pH dependence of NADH formation by dTDP-xylose, is 6.41. Lys164 and Thr134 are believed to play important roles in the stabilization of the anion of Tyr160 in a fashion similar to the roles of the corresponding residues in UDP-galactose 4 epimerase, which facilitate the ionization of Tyr149 in that enzyme [Liu, Y., et al. (1997) Biochemistry 35, 10675--10684]. Tyr160 is presumably the base for the first hydride transfer step, while Thr134 may relay a proton from the sugar to Tyr160. PMID- 11478887 TI - Functional inactivation of the human guanylyl cyclase C receptor: modeling and mutation of the protein kinase-like domain. AB - Receptor guanylyl cyclases possess an extracellular ligand-binding domain, a single transmembrane region, a region with sequence similar to that of protein kinases, and a C-terminal guanylyl cyclase domain. ATP regulates the activity of guanylyl cyclase C (GC-C), the receptor for the guanylin and stable toxin family of peptides, presumably as a result of binding to the kinase homology domain (KHD). Modeling of the KHD of GC-C indicated that it could adopt a structure similar to that of tyrosine kinases, and sequence comparison with other protein kinases suggested that lysine(516) was positioned in the KHD to interact with ATP. A monoclonal antibody GCC:4D7, raised to the KHD of GC-C, did not recognize ATP-bound GC-C, and its epitope mapped to a region in the KHD of residues 491- 568 of GC-C. Mutation of lysine(516) to an alanine in full-length GC-C (GC C(K516A)) dramatically reduced the ligand-stimulated activity of mutant GC-C, altered the ATP-mediated effects observed with wild-type GC-C, and failed to react with the GCC:4D7 monoclonal antibody. ATP interaction with wild-type GC-C converted a high-molecular weight oligomer of GC-C to a smaller sized oligomer. In contrast, GC-C(K516A) did not exhibit an alteration in its oligomeric status on incubation with ATP. We therefore suggest that the KHD in receptor guanylyl cyclases provides a critical structural link between the extracellular domain and the catalytic domain in regulation of activity in this family of receptors, and the presence of K(516) is critical for the possible proper orientation of ATP in this domain. PMID- 11478888 TI - Structures of ceftazidime and its transition-state analogue in complex with AmpC beta-lactamase: implications for resistance mutations and inhibitor design. AB - Third-generation cephalosporins are widely used beta-lactam antibiotics that resist hydrolysis by beta-lactamases. Recently, mutant beta-lactamases that rapidly inactivate these drugs have emerged. To investigate why third-generation cephalosporins are relatively stable to wild-type class C beta-lactamases and how mutant enzymes might overcome this, the structures of the class C beta-lactamase AmpC in complex with the third-generation cephalosporin ceftazidime and with a transition-state analogue of ceftazidime were determined by X-ray crystallography to 2.0 and 2.3 A resolution, respectively. Comparison of the acyl-enzyme structures of ceftazidime and loracarbef, a beta-lactam substrate, reveals that the conformation of ceftazidime in the active site differs from that of substrates. Comparison of the structures of the acyl-enzyme intermediate and the transition-state analogue suggests that ceftazidime blocks formation of the tetrahedral transition state, explaining why it is an inhibitor of AmpC. Ceftazidime cannot adopt a conformation competent for catalysis due to steric clashes that would occur with conserved residues Val211 and Tyr221. The X-ray crystal structure of the mutant beta-lactamase GC1, which has improved activity against third-generation cephalosporins, suggests that a tandem tripeptide insertion in the Omega loop, which contains Val211, has caused a shift of this residue and also of Tyr221 that would allow ceftazidime and other third generation cephalosporins to adopt a more catalytically competent conformation. These structural differences may explain the extended spectrum activity of GC1 against this class of cephalosporins. In addition, the complexed structure of the transition-state analogue inhibitor (K(i) 20 nM) with AmpC reveals potential opportunities for further inhibitor design. PMID- 11478889 TI - Structures of cytochrome c-549 and cytochrome c6 from the cyanobacterium Arthrospira maxima. AB - Cytochrome c(6) and cytochrome c-549 are small (89 and 130 amino acids, respectively) monoheme cytochromes that function in photosynthesis. They appear to have descended relatively recently from the same ancestral gene but have diverged to carry out very different functional roles, underscored by the large difference between their midpoint potentials of nearly 600 mV. We have determined the X-ray crystal structures of both proteins isolated from the cyanobacterium Arthrospira maxima. The two structures are remarkably similar, superimposing on backbone atoms with an rmsd of 0.7 A. Comparison of the two structures suggests that differences in solvent exposure of the heme and the electrostatic environment of the heme propionates, as well as in heme iron ligation, are the main determinants of midpoint potential in the two proteins. In addition, the crystal packing of both A. maxima cytochrome c-549 and cytochrome c(6) suggests that the proteins oligomerize. Finally, the cytochrome c-549 dimer we observe can be readily fit into the recently described model of cyanobacterial photosystem II. PMID- 11478890 TI - Aromatic substrate specificity of horseradish peroxidase C studied by a combined fluorescence line narrowing/energy minimization approach: the effect of localized side-chain reorganization. AB - Horseradish peroxidase C binds a wide variety of small H-donor compounds such as benzohydroxamic acid (BHA) and 2-naphthohydroxamic acid (NHA). In this work, we use the Mg(II)-mesoporphyrin prosthetic group derivative as a spectroscopic probe of the active site and of the interaction with the substrates. We report on high resolution fluorescence line-narrowed spectra which show that the effects of substrate binding on the electronic transitions are similar for both substrates and present data on the normal vibrational modes that are active in the vibronic spectra. Analysis of the vibrational frequencies shows that the Mg(II) ion is 5 coordinate in all cases, thus ruling out a solvent water as sixth ligand. The frequency shifts observed as a result of substrate binding are also indicative of a more rigid prosthetic group upon substrate binding. We present models for MgMP HRP and its complexes with both substrates and compare the resulting structures on the basis of a modeling approach combining energy minimization to finite difference Poisson--Boltzmann calculations which partitions the various relative protein contributions to substrate binding. We show that the electrostatic potential of the prosthetic group is modified by the binding event. Analysis of the unbound and bound energy-minimized structures shows that the enzyme modulates substrate binding by subtle charge reorganization in the vicinity of the catalytic site and that this rearrangement is not attributable to significant secondary structure conformational changes but to side-chain reorganization. PMID- 11478891 TI - Structural changes of pharaonis phoborhodopsin upon photoisomerization of the retinal chromophore: infrared spectral comparison with bacteriorhodopsin. AB - Archaeal rhodopsins possess a retinal molecule as their chromophores, and their light energy and light signal conversions are triggered by all-trans to 13-cis isomerization of the retinal chromophore. Relaxation through structural changes of the protein then leads to functional processes, proton pump in bacteriorhodopsin and transducer activation in sensory rhodopsins. In the present paper, low-temperature Fourier transform infrared spectroscopy is applied to phoborhodopsin from Natronobacterium pharaonis (ppR), a photoreceptor for the negative phototaxis of the bacteria, and infrared spectral changes before and after photoisomerization are compared with those of bacteriorhodopsin (BR) at 77 K. Spectral comparison of the C--C stretching vibrations of the retinal chromophore shows that chromophore conformation of the polyene chain is similar between ppR and BR. This fact implies that the unique chromophore-protein interaction in ppR, such as the blue-shifted absorption spectrum with vibrational fine structure, originates from both ends, the beta-ionone ring and the Schiff base regions. In fact, less planer ring structure and stronger hydrogen bond of the Schiff base were suggested for ppR. Similar frequency changes upon photoisomerization are observed for the C==N stretch of the retinal Schiff base and the stretch of the neighboring threonine side chain (Thr79 in ppR and Thr89 in BR), suggesting that photoisomerization in ppR is driven by the motion of the Schiff base like BR. Nevertheless, the structure of the K state after photoisomerization is different between ppR and BR. In BR, chromophore distortion is localized in the Schiff base region, as shown in its hydrogen out-of-plane vibrations. In contrast, more extended structural changes take place in ppR in view of chromophore distortion and protein structural changes. Such structure of the K intermediate of ppR is probably correlated with its high thermal stability. In fact, almost identical infrared spectra are obtained between 77 and 170 K in ppR. Unique chromophore-protein interaction and photoisomerization processes in ppR are discussed on the basis of the present infrared spectral comparison with BR. PMID- 11478892 TI - A novel protein involved in the functional assembly of the oxygen-evolving complex of photosystem II in Synechocystis sp. PCC 6803. AB - Mutation of Glu69 to Gln in the D2 protein of photosystem II is known to lead to a loss of photoautotrophic growth in Synechocystis sp. PCC 6803. However, second site mutants (pseudorevertants) with restored photoautotrophic growth but still maintaining the E69Q mutation in D2 are easily obtained. Using a genomic mapping technique involving functional complementation, the secondary mutation was mapped to slr0286 in two independent mutants. The mutations in Slr0286 were R42M or R394H. To study the function of Slr0286, mutants of E69Q and of the wild-type strain were made that lacked slr0286. Deletion of slr0286 did not affect photoautotrophic capacity in wild type but led to a marked decrease in the apparent affinity of Ca(2+) to its binding site at the water-splitting system of photosystem II and to a reduced heat tolerance of the oxygen-evolving system, particularly in E69Q. Moreover, a small increase in the half-time for photoactivation of the oxygen-evolving complex of photosystem II for both wild type and the E69Q mutant was observed in the absence of Slr0286. The accumulation of photosystem II reaction centers, dark stability of the oxygen-evolving apparatus, stability of oxygen evolution, and the kinetics of charge recombination between Q(A)(-) and the donor side were not affected by deletion of slr0286. Slr0286 lacks clear functional motifs, and no homologues are apparent in other organisms, even not in other cyanobacteria. In any case, Slr0286 appears to help the functional assembly and stability of the water-splitting system of photosystem II. PMID- 11478893 TI - In vitro activation of soluble guanylyl cyclase and nitric oxide release: a comparison of NO donors and NO mimetics. AB - Nitric oxide (NO) performs a central role in biological systems, binding to the heme site of soluble guanylyl cyclase (sGC), leading to enzyme activation and elevation of intracellular levels of cGMP. Organic nitrates, in particular, nitroglycerin (GTN), are clinically important nitrovasodilators that function as NO-mimetics in biological systems. Comparison of sGC activation data with electrochemically measured rates of NO release for genuine NO donors, NONOates and nitrosothiols, yields an excellent correlation between the EC(50) for sGC activation and the rate constant for NO release, k(NO). However, activation of sGC by GTN and the nitrates has very different characteristics, including the requirement for specific added thiols, for example, cysteine. The reaction of GTN with cysteine in anaerobic solution yields NO slowly, and NO release, measured by chemiluminescence detection, is quenched by added metal ion chelator. The generation of NO under aerobic conditions is 100-fold slower than the anaerobic reaction. Furthermore, NO release from the reaction of GTN with cysteine in phosphate buffer is too slow to account for sGC activation by GTN/cysteine. The slow rate of the chemical reaction to release NO suggests that nitrates can activate sGC by an NO-independent mechanism. In contrast to the genuine NO donors, GTN behaves as a partial agonist with respect to sGC activation, but in the presence of the allosteric sGC activator, YC-1, GTN exhibits full agonist activity. PMID- 11478894 TI - Site-directed mutations at D1-His198 and D2-His197 of photosystem II in Synechocystis PCC 6803: sites of primary charge separation and cation and triplet stabilization. AB - Site-directed mutations were introduced to replace D1-His198 and D2-His197 of the D1 and D2 polypeptides, respectively, of the photosystem II (PSII) reaction center of Synechocystis PCC 6803. These residues coordinate chlorophylls P(A) and P(B) which are homologous to the special pair Bchlorophylls of the bacterial reaction centers that are coordinated respectively by histidines L-173 and M-200 (202). P(A) and P(B) together serve as the primary electron donor, P, in purple bacterial reaction centers. In PS II, the site-directed mutations at D1 His198 affect the P(+)--P-absorbance difference spectrum. The bleaching maximum in the Soret region (in WT at 433 nm) is blue-shifted by as much as 3 nm. In the D1 His198Gln mutant, a similar displacement to the blue is observed for the bleaching maximum in the Q(y) region (672.5 nm in WT at 80 K), whereas features attributed to a band shift centered at 681 nm are not altered. In the Y(Z*)--Y(Z) difference spectrum, the band shift of a reaction center chlorophyll centered in WT at 433--434 nm is shifted by 2--3 nm to the blue in the D1-His198Gln mutant. The D1-His198Gln mutation has little effect on the optical difference spectrum, (3)P--(1)P, of the reaction center triplet formed by P(+)Pheo(-) charge recombination (bleaching at 681--684 nm), measured at 5--80 K, but becomes visible as a pronounced shoulder at 669 nm at temperatures > or =150 K. Measurements of the kinetics of oxidized donor--Q(A)(-) charge recombination and of the reduction of P(+) by redox active tyrosine, Y(Z), indicate that the reduction potential of the redox couple P(+)/P can be appreciably modulated both positively and negatively by ligand replacement at D1-198 but somewhat less so at D2-197. On the basis of these observations and others in the literature, we propose that the monomeric accessory chlorophyll, B(A), is a long-wavelength trap located at 684 nm at 5 K. B(A)* initiates primary charge separation at low temperature, a function that is increasingly shared with P(A)* in an activated process as the temperature rises. Charge separation from B(A)* would be potentially very fast and form P(A)(+)B(A)(-) and/or B(A)(+)Pheo(-) as observed in bacterial reaction centers upon direct excitation of B(A) (van Brederode, M. E., et al. (1999) Proc. Natl. Acad Sci. 96, 2054--2059). The cation, generated upon primary charge separation in PSII, is stabilized at all temperatures primarily on P(A), the absorbance spectrum of which is displaced to the blue by the mutations. In WT, the cation is proposed to be shared to a minor extent (approximately 20%) with P(B), the contribution of which can be modulated up or down by mutation. The band shift at 681 nm, observed in the P(+)-P difference spectrum, is attributed to an electrochromic effect of P(A)(+) on neighboring B(A). Because of its low-energy singlet and therefore triplet state, the reaction center triplet state is stabilized on B(A) at < or =80 K but can be shared with P(A) at >80 K in a thermally activated process. PMID- 11478895 TI - Kinetics of charge separation and A0- --> A1 electron transfer in photosystem I reaction centers. AB - The charge separation P700*A(0) --> P700(+)A(0)(-) and the subsequent electron transfer from the primary to secondary electron acceptor have been studied by subtracting absorption difference profiles for cyanobacterial photosystem I (PS I) complexes with open and closed reaction centers. Samples were excited at 660 nm, which lies toward the blue edge of the core antenna absorption spectrum. The resulting PS I kinetics were analyzed in terms of the relevant P700, P700(+), A(0), and A(0)(-) absorption spectra. In our kinetic model, the radical pair P700(+)A(0)(-) forms with 1.3 ps rise kinetics after creation of electronically excited P700*. The formation of A(1)(-) via electron transfer from A(0)(-) requires approximately 13 ps. The kinetics of the latter step are appreciably faster than previously estimated by other groups (20--50 ps). PMID- 11478896 TI - Characterization of the binding of deuteroporphyrin IX to the magnesium chelatase H subunit and spectroscopic properties of the complex. AB - Magnesium protoporphyrin chelatase catalyzes the insertion of a Mg(2+) ion into protoporphyrin IX, which can be considered as the first committed step of (bacterio)chlorophyll synthesis. In the present work, the Mg chelatase H subunits from both Synechocystis and Rhodobacter sphaeroides were studied because of the differing requirements of these organisms for modified cyclic tetrapyrroles. Deuteroporphyrin was shown to be a substrate for Mg chelatase. Analytical HPLC gel filtration was used to show that an H-deuteroporphyrin complex can be reconstituted by incubating the magnesium chelatase H subunit with a molar excess of deuteroporphyrin and that these complexes are monomers. The binding process occurs in the absence of Mg(2+) or ATP or the I or D subunits of Mg chelatase. The emission from Trp residues in the H subunit is partly quenched when deuteroporphyrin is bound. Quantitative analysis of Trp fluorescence quenching led to determination of the K(d) values for deuteroporphyrin binding to BchH from Rb. sphaeroides and ChlH from Synechocystis, which are 1.22 +/- 0.42 microM and 0.53 +/- 0.12 microM for ChlH and BchH, respectively. In the case of ChlH, but not BchH, the K(d) increased 4-fold in the presence of MgATP(2-). Red shifts in absorbance and excitation peaks were observed in the B band of the bound porphyrin in comparison with deuteroporphyrin in solution, as well as reduced yield and red shifts of up to 8 nm in fluorescence emission. These alterations are consistent with a slightly deformed nonplanar conformation of the bound porphyrin. Mg deuteroporphyrin, the product of the Mg chelation reaction, was shown to form a complex with either ChlH or BchH; in each case the K(d) for Mg deuteroporphyrin is similar to that for deuteroporphyrin. The implications of the H-Mg protoporphyrin interaction for the next enzyme in the chlorophyll biosynthetic pathway, Mg protoporphyrin methyltransferase, are discussed. PMID- 11478897 TI - SELEX selection of high-affinity oligonucleotides for bacteriophage Ff gene 5 protein. AB - The Ff gene 5 protein (g5p) is a cooperative ssDNA-binding protein. SELEX was used to identify DNA sequences favorable for g5p binding at physiological ionic strength (200 mM NaCl) and 37 degrees C. Sequences were selected from a library of 58-mers that contained a central variable segment of 26 nucleotides. DNA sequences selected after eight rounds of SELEX were mostly G-rich, with multiple copies of CPuGGPy, TPuGGGPy, and/or PyPuPuGGGPy motifs. This was unexpected, since g5p has higher binding affinities for polypyrimidine than for polypurine sequences. The most recurrent G-rich sequence, named I-3, was found to have g5p binding properties that were correlated with a structural transition. At 10 mM NaCl, I-3 existed in a single-stranded form that was saturated by g5p in an all or-none fashion. At 200 mM NaCl, I-3 existed in a structured form that showed CD spectral features of G-quadruplexes. The g5p binding affinity for this structured form of I-3 was >100-fold higher than for the single-stranded form. Moreover, the structured I-3 was saturated by g5p in two steps, the first of which was the formation of an apparent initiation complex consisting of one I-3 strand and about three g5p dimers. Nuclease S1 footprinting and other experiments showed that g5p molecules in the initiation complex at 200 mM NaCl were bound directly to the G-rich variable segment and that the structure of I-3 was retained after saturation by g5p. Thus, G-rich motifs may form structures favorable for initiation of g5p binding and also provide the actual g5p-binding sites. PMID- 11478898 TI - Monomer-dimer equilibrium and oxygen binding properties of ferrous Vitreoscilla hemoglobin. AB - The monomer-dimer equilibrium and the oxygen binding properties of ferrous recombinant Vitreoscilla hemoglobin (Vitreoscilla Hb) have been investigated. Sedimentation equilibrium data indicate that the ferrous deoxygenated and carbonylated derivatives display low values of equilibrium dimerization constants, 6 x 10(2) and 1 x 10(2) M(-1), respectively, at pH 7.0 and 10 degrees C. The behavior of the oxygenated species, as measured in sedimentation velocity experiments, is superimposable to that of the carbonylated derivative. The kinetics of O(2) combination, measured by laser photolysis at pH 7.0 and 20 degrees C, is characterized by a second-order rate constant of 2 x 10(8) M(-1) s( 1) whereas the kinetics of O(2) release at pH 7.0 is biphasic between 10 and 40 degrees C, becoming essentially monophasic below 10 degrees C. Values of the first-order rate constants (at 20 degrees C) and of the activation energies for the fast and slow phases of the Vitreoscilla Hb deoxygenation process are 4.2 s( 1) and 19.2 kcal mol(-1) and 0.15 s(-1) and 24.8 kcal mol(-1), respectively. Thus the biphasic kinetics of Vitreoscilla Hb deoxygenation is unrelated to the association state of the protein. The observed biphasic oxygen release may be accounted for by the presence of two different conformers in thermal equilibrium within the monomer. The two conformers may be assigned to a structure in which the heme-iron-bound ligand is stabilized by direct hydrogen bonding to TyrB10 and a structure in which such interaction is absent. The slow interconversion between the two conformers may reflect a very large conformational rearrangement in the disordered distal pocket segment connecting helices C and E. PMID- 11478899 TI - Mg2+-linked oligomerization modulates the catalytic activity of the Lon (La) protease from Mycobacterium smegmatis. AB - Lon (La) proteases are multimeric enzymes that are activated by ATP and Mg(2+) ions and stimulated by unfolded proteins such as alpha-casein. The peptidase activity of the Lon protease from Mycobacterium smegmatis (Ms-Lon) is dependent upon both its concentration and that of Mg(2+). Addition of alpha-casein partially substitutes for Mg(2+) in activating the enzyme. In chemical dissociation experiments, higher concentrations of urea were required to inhibit Ms-Lon's catalytic activities after an addition of alpha-casein. Analytical ultracentrifugation was used to directly probe the effect of activators of peptidase activity on Ms-Lon self-association. Sedimentation velocity experiments reveal that Ms-Lon monomers are in a reversible equilibrium with oligomeric forms of the protein and that the self-association reaction is facilitated by Mg(2+) ions but not by AMP-PNP or ATP gamma S. NaCl at 100 mM facilitates oligomerization and stimulates peptidase activity at suboptimal concentrations of MgCl(2). Sedimentation equilibrium analysis shows that Ms-Lon associates to a hexamer at 50 mM Tris and 10 mM MgCl(2), at pH 8.0 and 20 degrees C, and that the assembly reaction is Mg(2+) dependent; the mole fraction of hexamer decreases with decreasing MgCl(2) to undetectable levels in 10 mM EDTA. The analysis of experiments conducted at a series of initial protein and MgCl(2) concentrations yields two assembly models: dimer <--> tetramer <--> hexamer and timer <--> hexamer, equally consistent with the data. Limited trypsin digestion, CD, and tryptophan fluorescence suggest only minor changes in secondary and tertiary structure upon Mg(2+)-linked oligomerization. These results show that activation of Ms-Lon peptidase activity requires oligomerization and that Ms-Lon self association reaction is facilitated by its activator, Mg(2+), and stimulator, unfolded protein. PMID- 11478900 TI - Probing inhibitor-induced conformational changes along the interface between tissue factor and factor VIIa. AB - Upon injury of a blood vessel, activated factor VII (FVIIa) forms a high-affinity complex with its allosteric regulator, tissue factor (TF), and initiates blood clotting. Active site-inhibited factor VIIa (FVIIai) binds to TF with even higher affinity. We compared the interactions of FVIIai and FVIIa with soluble TF (sTF). Six residues in sTF were individually selected for mutagenesis and site-directed labeling. The residues are distributed along the extensive binding interface, and were chosen because they are known to interact with the different domains of FVIIa. Fluorescent and spin probes were attached to engineered Cys residues to monitor local changes in hydrophobicity, accessibility, and rigidity in the sTF- FVIIa complex upon occupation of the active site of FVIIa. The results show that inhibition of FVIIa caused the structures around the positions in sTF that interact with the protease domain of FVIIa to become more rigid and less accessible to solvent. Thus, the presence of an active site inhibitor renders the interface in this region less flexible and more compact, whereas the interface between sTF and the light chain of FVIIa is unaffected by active site occupancy. PMID- 11478901 TI - Time-dependent inhibition of protein farnesyltransferase by a benzodiazepine peptide mimetic. AB - Protein farnesyltransferase (FTase) and protein geranylgeranyltransferase-I (GGTase-I) catalyze the prenylation of proteins with a carboxy-terminal tetrapeptide sequence called a CaaX box, where C refers to cysteine, "a" refers to an aliphatic residue, and X typically refers to methionine, serine, or glutamine (FTase), or to leucine (GGTase-I). Marsters and co-workers [(1994) Bioorg. Med. Chem. 2, 949--957] developed inhibitors of FTase with cysteine and methionine attached to an inner hydrophobic benzodiazepine scaffold. We found that the most potent of these compounds (BZA-2B) resulted in the time-dependent inhibition of FTase. The K(i) of BZA-2B for FTase, which is the dissociation constant of the initial complex, was 79 +/- 13 nM, and the K(i)*, which is the overall dissociation of inhibitor for all enzyme forms, was 0.91 +/- 0.12 nM. The first-order rate constant for the conversion of the initial complex to the final complex was 1.4 +/- 0.2 min(-1), and that for the reverse process was 0.016 +/- 0.002 min(-1). The latter rate constant corresponds to a half-life of the final complex of 45 min. Our experiments favor the notion that the inhibitor binds to the FTase--farnesyl diphosphate complex which then undergoes an isomerization to form a tighter FTase*--farnesyl diphosphate--BZA2-B complex. Diazepam, a compound with a benzodiazepine nucleus but lacking amino acid extensions, was a weak (K(i) = 870 microM) but not time-dependent inhibitor of FTase. Cys-Val-Phe-Met and Cys 4-aminobenzoyl-Met were instantaneous and not time-dependent inhibitors of FTase. Furthermore, BZA-4B, with a leucine specificity determinant, was a classical competitive inhibitor of GGTase-I and not a time-dependent inhibitor. PMID- 11478902 TI - Stability and function of interdomain linker variants of glucoamylase 1 from Aspergillus niger. AB - Several variants of glucoamylase 1 (GA1) from Aspergillus niger were created in which the highly O-glycosylated peptide (aa 468--508) connecting the (alpha/alpha)(6)-barrel catalytic domain and the starch binding domain was substituted at the gene level by equivalent segments of glucoamylases from Hormoconis resinae, Humicola grisea, and Rhizopus oryzae encoding 5, 19, and 36 amino acid residues. Variants were constructed in which the H. resinae linker was elongated by proline-rich sequences as this linker itself apparently was too short to allow formation of the corresponding protein variant. Size and isoelectric point of GA1 variants reflected differences in linker length, posttranslational modification, and net charge. While calculated polypeptide chain molecular masses for wild-type GA1, a nonnatural proline-rich linker variant, H. grisea, and R. oryzae linker variants were 65,784, 63,777, 63,912, and 65,614 Da, respectively, MALDI-TOF-MS gave values of 82,042, 73,800, 73,413, and 90,793 Da, respectively, where the latter value could partly be explained by an N-glycosylation site introduced near the linker C-terminus. The k(cat) and K(m) for hydrolysis of maltooligodextrins and soluble starch, and the rate of hydrolysis of barley starch granules were essentially the same for the variants as for wild-type GA1. beta-Cyclodextrin, acarbose, and two heterobidentate inhibitors were found by isothermal titration calorimetry to bind to the catalytic and starch binding domains of the linker variants, indicating that the function of the active site and the starch binding site was maintained. The stability of GA1 linker variants toward GdnHCl and heat, however, was reduced compared to wild-type. PMID- 11478903 TI - Mutational analysis of the thyrotropin-releasing hormone-degrading ectoenzyme. similarities and differences with other members of the M1 family of aminopeptidases and thermolysin. AB - Thyrotropin-releasing hormone-degrading ectoenzyme (TRH-DE) is a TRH-specific peptidase which catalyzes the inactivation of the peptidergic signal substance TRH. As indicated by sequence alignment, TRH-DE and the other members of the M1 family of aminopeptidases have a distinct set of conserved amino acid residues in common. By replacing amino acid residues that are putatively involved in catalysis, we could demonstrate that the enzymatic activities of the mutants E408D, E442D, E464Q, E464D, Y528F, H507R, and H507F are dramatically decreased, essentially due to the changes of V(max). The mutant enzymes E408Q and E442Q are inactive, whereas the specific enzymatic activity of the mutants R488Q, R488A, and Y554F are similar to that of the wild-type enzyme. These data strongly suggest that E408, E442, Y528, and H507 are involved in the catalytic process of TRH-DE while E464 presumably represents the third zinc-coordinating residue and may be equivalent to E166 in thermolysin. In contrast, amino acid residues R488 and Y554 seem not to be involved in the catalytic mechanism of TRH-DE. PMID- 11478904 TI - Classical and slow-binding inhibitors of human type II arginase. AB - Arginases catalyze the hydrolysis of L-arginine to yield L-ornithine and urea. Recent studies indicate that arginases, both the type I and type II isozymes, participate in the regulation of nitric oxide production by modulating the availability of arginine for nitric oxide synthase. Due to the reciprocal regulation between arginase and nitric oxide synthase, arginase inhibitors have therapeutic potential in treating nitric oxide-dependent smooth muscle disorders, such as erectile dysfunction. We demonstrate the competitive inhibition of the mitochondrial human type II arginase by N(omega)-hydroxy-L-arginine, the intermediate in the reaction catalyzed by nitric oxide synthase, and its analogue N(omega)-hydroxy-nor-L-arginine, with K(i) values of 1.6 microM and 51 nM at pH 7.5, respectively. We also demonstrate the inhibition of human type II arginase by the boronic acid-based transition-state analogues 2(S)-amino-6-boronohexanoic acid (ABH) and S-(2-boronoethyl)-L-cysteine (BEC), which are known inhibitors of type I arginase. At pH 7.5, both ABH and BEC are classical, competitive inhibitors of human type II arginase with K(i) values of 0.25 and 0.31 microM, respectively. However, at pH 9.5, ABH and BEC are slow-binding inhibitors of the enzyme with K(i) values of 8.5 and 30 nM, respectively. The findings presented here indicate that the design of arginine analogues with uncharged, tetrahedral functional groups will lead to the development of more potent inhibitors of arginases at physiological pH. PMID- 11478905 TI - A kinetic model to evaluate cholesterol efflux from THP-1 macrophages to apolipoprotein A-1. AB - The kinetics (0 to 3 h) of cholesterol efflux to delipidated apolipoprotein A-1 were investigated, and the experimental data were best fitted to a mathematical model that involves two independent pathways of cholesterol efflux. The first pathway with a rate constant of 4.6 h(-1) is fast but removes only 3-5% of total cholesterol. After preconditioning apoA-1, it was found that this pathway remains, and hence it is a property of the cholesterol-loaded cells rather than due to modification on the apolipoprotein. This fast initial efflux does not seem to contribute to cholesterol efflux at later stages (>1 h) where a second pathway predominates. However, the fast initial efflux pool can be restored if apoA-1 is withdrawn. The second slower pathway (k(membrane--media) = 0.79 h(-1)) is associated with cholesterol ester hydrolysis whose rate constant could be experimentally verified (k(cal) = 0.43, k(exp) = 0.38 +/- 0.05). The model suggests that two different plasma membrane domains are involved in the two pathways. Loading of the cells with an oxysterol, 7-ketocholesterol (7K), inhibits efflux from both pathways. The model predicts that 7K decreases the initial efflux by decreasing the available cholesterol (by possibly affecting lipid packing), while all rate constants in the second pathway are decreased. In conclusion, the kinetic model suggests that cholesterol efflux to apoA-1 is a two step process. In the first step, some of the plasma membrane cholesterol contributes to a fast initial efflux (possibly from lipid rafts) and leads to a second pathway that mobilizes intracellular cholesterol mobilization. PMID- 11478906 TI - Differential roles of developmentally distinct SNAP-25 isoforms in the neurotransmitter release process. AB - The role of SNAP-25 (synaptosomal associated protein of 25 kDa) isotypes in the neurotransmitter release process was examined by varying their relative abundance during PC12 cell differentiation induced by nerve growth factor (NGF). Norepinephrine release by NGF-differentiated PC12 cells is more sensitive to type A botulinum toxin (BoNT/A) than by nondifferentiated cells, while both differentiated and nondifferentiated PC12 cells are equally sensitive to type E botulinum toxin (BoNT/E). The differential sensitivity to BoNT/A corresponds to an altered susceptibility of SNAP-25 isotypes to BoNT/A cleavage in vitro, whereas both isotypes are equally vulnerable to cleavage by BoNT/E. Using recombinant SNAP-25 preparations, we show that BoNT/A cleaves SNAP-25b (present in differentiated cells) 2-fold more readily than SNAP-25a (present in both differentiated and nondifferentiated cells). Structural studies using far ultraviolet circular dichroism (UV--CD) and thermal denaturation suggest a difference in the polypeptide folding as the underlying molecular basis for the differential sensitivity of SNAP-25b and SNAP-25a to BoNT/A cleavage. We propose differential roles for SNAP-25b and SNAP-25a in the neurotransmitter release process since our results suggest that BoNT/A inhibits neurotransmitter release by primarily cleaving SNAP-25b. PMID- 11478907 TI - Short segment of human melanin-concentrating hormone that is sufficient for full activation of human melanin-concentrating hormone receptors 1 and 2. AB - Human melanin-concentrating hormone (hMCH) is a potent but nonselective agonist at human melanin-concentrating hormone receptors 1 and 2 (hMCH-1R and hMCH-2R, respectively). To determine the structural features of this neuropeptide which are necessary for efficient binding to and activation of the receptors, Ala substituted, open-chain, and truncated analogues were synthesized and tested in the binding assays in CHO cells expressing hMCH-1R and hMCH-2R, and in functional assays measuring the level of intracellular calcium mobilization in human HEK-293 cells expressing these receptors. A compound consisting merely of the cyclic core of hMCH with the Arg attached to the N-terminus of the disulfide ring was found to activate both hMCH-1R and hMCH-2R about as effectively as full-length hMCH. Thus, the sequence Arg-cyclo(S-S)(Cys-Met-Leu-Gly-Arg-Val-Tyr-Arg-Pro-Cys) appears to constitute the "active core" that is necessary for agonist potency at hMCH-1R and hMCH-2R. A potent and approximately 4-fold more selective agonist at hMCH-1R than at hMCH-2R is also reported. PMID- 11478908 TI - A molecular beacon strategy for the thermodynamic characterization of triplex DNA: triplex formation at the promoter region of cyclin D1. AB - We studied the formation of triplex DNA in the purine-pyrimidine-rich promoter site sequence of cyclin D1, located at -116 to -99 from the transcription initiation site, with a molecular beacon comprised of a G-rich 18-mer triplex forming oligodeoxyribonucleotide. Formation of triplex DNA was monitored by enhanced fluorescence of the beacon, due to the weakening of fluorescence energy transfer, upon its binding to the target duplex. Electrophoretic mobility shift assay confirmed triplex DNA formation by these oligonucleotides. In low salt buffer (10 mM Na(+)), triplex DNA formation was not observed in the absence of a ligand such as spermine. At room temperature (22 degrees C), the equilibrium association constant (K(a)) calculated in the presence of 1 microM spermine and 10 mM Na(+) was 3.2 x 10(8) M(-1). The K(a) value was 1.0 x 10(9) M(-1) in the presence of 150 mM Na(+), and it increased by 10-fold by the addition of 1 mM spermine. Delta H, Delta S, and Delta G of triplex DNA formation, calculated from the temperature dependence of K(a) in the range of 20--45 degrees C, were -35.9 kcal/mol, -77 cal/(mol.K), and -13 kcal/mol, respectively, in the presence of 150 mM NaCl. The corresponding values were -52.9 kcal/mol, -132.5 cal/(mol.K), and 13.4 kcal/mol in the presence of 150 mM NaCl and 1 mM spermine. Structurally related polyamines exerted different degrees of triplex DNA stabilization, as determined by binding constant measurements. Comparison of spermine versus hexamine showed a 17-fold increase in the equilibrium association constant, whereas bis(ethyl) derivatization lead to a 4-fold decrease of this value. In the absence of added duplex and polyamines, the molecular beacon dissociated with a melting temperature of 67 degrees C. Thermodynamic parameters of beacon melting were calculated from the melting curve, and the Delta H, Delta S, and Delta G values were 37.8 kcal/mol, 112 cal/(mol.K), and 4.4 kcal/mol, respectively. These results demonstrate that molecular beacons can be used for the direct determination of the equilibrium association constants and thermodynamic parameters of triplex DNA formation in the presence of ligands such as polyamines. PMID- 11478909 TI - pH and cation effects on the properties of parallel pyrimidine motif DNA triplexes. AB - The effects of cytosine protonation and various cations on the properties of parallel pyrimidine motif DNA triplexes were intensively investigated and characterized by several different techniques, such as circular dichroism (CD) conformation, ultraviolet (UV) melting, differential scanning calorimetry (DSC) thermal denaturation, and surface plasmon resonance (SPR) real-time dynamics. The comparative CD spectra of the triplex and the corresponding homoduplexes showed that the negative peak at approximately 218 nm would be the eigenpeak of the Hoogsteen paired strand, and moreover, the formation pathway of a triplex was significantly pH-dependent and fell into three groups: under acidic conditions, the triplex is formed by a one-step docking, under near physiological conditions, the Watson-Crick duplex is first structured and then accepts the Hoogsteen third strand into its major groove, and under basic conditions, the triplex is not formed. The pH-dependent thermodynamics of the global triplex, the Watson-Crick antiparallel duplex, and the Crick-Hoogsteen parallel duplex were comparatively discussed for the first time. These data revealed that the thermodynamic stabilities of the Watson-Crick-Hoogsteen triplex and the Crick-Hoogsteen duplex would be strongly dependent on cytosine protonation, but a low-pH environment somewhat destabilized the Watson-Crick duplex. The binding energy of triplex formation would be different from the unfolding energy of triplex melting under acidic conditions due to the disparity in the pathway between the formation and unfolding of a triplex. Real-time dynamic measurements showed that the association and dissociation rate constants of a duplex-to-triplex formation are (1.98 +/- 0.24) x 10(3) M(-1) s(-1) and (4.09 +/- 0.96) x 10(-4) s(-1) at 20 degrees C and pH 6.0, respectively. The formation energy of the duplex-to-triplex transition derived from SPR measurements was in agreement with the unfolding energy of the free Hoogsteen paired duplex derived from UV measurements. The calorimetric enthalpies of the triplex-to-duplex-to-single transition were 39.3 and 75.3 kcal/mol under near physiological conditions (pH 7.0), respectively, which were underestimated relative to the van't Hoff enthalpies. In addition, the effects of various cations, ionic strength, mixed-valent cations, and the position of the C(+)xG.C triplets on the thermodynamics of the triplexes were addressed under near physiological conditions. The interaction of metal ions with the triplexes clearly depended on the type and ionic strength of the cations, and the efficiency with which the cations stabilized the global triplex was in the order Mg(2+) > Mn(2+) > Ca(2+) > Ba(2+) >> Na(+). These observations would be useful for the design of triplex-forming oligonucleotides for antigene drugs and therapeutic purposes. PMID- 11478910 TI - Magnesium increases the curvature of duplex DNA that contains dA tracts. AB - Distinct structural features of DNA, such as the curvature of dA tracts, are important in the recognition, packaging, and regulation of DNA. Physiologically relevant concentrations of magnesium have been found to enhance the curvature of dA tract DNAs, as monitored by solution-state NMR, indicating that the structure of DNA depends on the cations present in solution. A model is presented which accounts for the sequence-dependent effects of magnesium on DNA curvature as well as for the previously known sequence-independent effect on DNA flexibility. PMID- 11478911 TI - Effect of divalent metal cations on the dimerization of OXA-10 and -14 class D beta-lactamases from Pseudomonas aeruginosa. AB - The factors influencing the oligomerization state of OXA-10 and OXA-14 class D beta-lactamases in solution have been investigated. Both enzymes were found to exist as an equilibrium mixture of a monomer and dimer, with a K(d) close to 40 microM. The dimeric form was stabilized by divalent metal cations. The ability of different metal ions to stabilize the dimer was in the following order: Cd(2+) > Cu(2+) > Zn(2+) > Co(2+) > Ni(2+) > Mn(2+) > Ca(2+) > Mg(2+). The apparent K(d)s describing the binding of Zn(2+) and Cd(2+) cations to the OXA-10 dimer were 7.8 and 5.7 microM, respectively. The metal ions had a profound effect on the thermal stability of the protein complex observed by differential scanning calorimetry. The enzyme showed a sharp transition with a T(m) of 58.7 degrees C in the absence of divalent cations, and an equally sharp transition with a T(m) of 78.4 degrees C in the presence of a saturating concentration of the divalent cation. The thermal transition observed at intermediate concentrations of divalent metal ions was rather broad and lies between these two extremes of temperature. The equilibrium between the monomer and dimer is dependent on pH, and the optimum for the formation of the dimer shifted from pH 6.0 in the absence of divalent cations to pH 7.5 at saturating concentrations. The beta-lactamase activity increased approximately 2-fold in the presence of saturating concentrations of zinc and cadmium ions. Reaction with beta-lactams caused a shift in the equilibrium toward monomer formation, and thus an apparent inactivation, but the divalent cations protected against this effect. PMID- 11478912 TI - Evidence that Gal11 protein is a target of the Gal4 activation domain in the mediator. AB - The mediator is an approximately 20 protein complex that is essential for the transcription of most genes in yeast. It is contacted by a number of gene specific activators, but the details of these interactions are not well understood in most cases. Here, evidence is presented that the mediator component Gal11 represents at least one target of the Gal4 activation domain (AD). Deletion of Gal11 is shown to decrease the affinity of the Gal4 AD for the mediator, and direct binding of an N-terminal domain of Gal11 with the Gal4 AD is demonstrated. Quantitative studies, however, indicate that the K(D) of the 1:1 Gal4 AD--Gal11 complex is modest. Combined with in vivo data showing that Delta gal11 cells exhibit reduced, but still significant, Gal4-mediated gene expression, these results suggest that the dimeric activator might also contact another protein in the mediator in addition to Gal11. PMID- 11478913 TI - 15N and 31P solid-state NMR investigations on the orientation of zervamicin II and alamethicin in phosphatidylcholine membranes. AB - The topologies of zervamicin II and alamethicin, labeled with (15)N uniformly, selectively, or specifically, have been investigated by oriented proton-decoupled (15)N solid-state NMR spectroscopy. Whereas at lipid-to-peptide (L/P) ratios of 50 (wt/wt) zervamicin II exhibits transmembrane alignments in 1,2-dicapryl (di C10:0-PC) and 1,2-dilauroyl (di-C12:0-PC) phosphatidylcholine bilayers, it adopts orientations predominantly parallel to the membrane surface when the lengths of the fatty acyl chains are extended. The orientational order of zervamicin II increases with higher phospholipid concentrations, and considerable line narrowing is obtained in di-C10:0-PC/zervamicin II membranes at L/P ratios of 100 (wt/wt). In contrast to zervamicin, alamethicin is transmembrane throughout most, if not all, of its length when reconstituted into 1-palmitoyl-2-oleoyl-sn-glycero 3-phosphocholine bilayers. The (31)P solid-state NMR spectra of all phospholipid/peptaibol samples investigated show a high degree of headgroup order, indicating that the peptides do not distort the bilayer structure. The observed differences in peptide orientation between zervamicin and alamethicin are discussed with reference to differences in their lengths, helical conformations, distribution of (hydroxy)proline residues, and hydrophobic moments. Possible implications for peptaibol voltage-gating are also described. PMID- 11478914 TI - Inhibition and stimulation of phospholipid scrambling activity. Consequences for lipid asymmetry, echinocytosis, and microvesiculation of erythrocytes. AB - An increase of the intracellular Ca(2+) concentration in erythrocytes is known to activate rapid nonspecific bidirectional translocation of membrane-inserted phospholipid probes and to decrease the asymmetric distribution of endogenous membrane phospholipids. These scrambling effects are now shown to be suppressed by pretreatment of cells with the essentially impermeable reagents 4,4' diisothiocyanostilbene-2,2'-disulfonic acid and 2,4,6-trinitrobenzenesulfonate. The inhibitory effects are no longer observed during renewed activation of scrambling following a first transient activation by Ca(2+). Assuming the involvement of the human scramblase, this suggests a conformational alteration of this protein during activation by Ca(2+). Marked suppression of scrambling activity is also observed in cells pretreated with the disulfide reducing agent dithioerythritol which can be reverted by the SH oxidizing agent diamide. This indicates the importance of intramolecular and/or intersubunit disulfide bonds for the function of the scramblase. On the other hand, treatment of cells with the SH reagents N-ethylmaleimide and phenylarsine oxide enhances Ca(2+)-activated scrambling and diminution of asymmetry of membrane phospholipids. This suggests an allosteric connection of several protein SH groups to the translocation mechanism. The inhibitors retain their strong suppressive effects. Besides covalent modification, addition of oligomycin highly stimulates and addition of clotrimazole suppresses the Ca(2+)-activated translocation. No evidence for a role of the ATP-binding cassette transporter ABCA1 in the Ca(2+)-activated outward translocation is obtained. Suppression of phospholipid scrambling by dithioerythritol inhibits Ca(2+)-induced spheroechinocytosis and reduces the extent of subsequent microvesiculation. Scrambling of endogenous phospholipids is proposed to induce echinocytosis and to have only a stimulatory effect on microvesiculation. PMID- 11478915 TI - Expression of the acidic stretch of nardilysin as a functional binding domain. AB - Kinetic evidence suggests an acidic region in nardilysin binds polyamines and acts as a regulatory domain. The binding of approximately 5 mol of spermine/mol of nardilysin was demonstrated. The binding curve was sigmoidal exhibiting an IC(50) of approximately 118 microM and a Hill coefficient of 1.8. Spermine diminished the tryptophan fluorescence of the enzyme and increased its sensitivity to protease V8. The acidic stretch from mouse and human nardilysin were expressed as glutathione transferase fusion proteins. All fusion proteins bound spermine with an IC(50) of 40 to 110 microM. The mouse fusion protein bound approximately 7 mol of spermine exhibiting a sigmoidal binding curve and a Hill coefficient of 1.4. The human acidic stretch, containing fewer acidic residues, bound approximately 5 mol of spermine/mol with a hyperbolic binding curve. Chimeric fusion proteins containing the N-terminus of the mouse acidic region fused to the C-terminus of the human acidic region bound approximately 10 mol of spermine, while the opposite chimera bound approximately 4 mol of spermine/mol. The N-terminal region of the mouse acidic domain binds 3--4 mol spermine/mol exhibiting a Hill coefficient of 1.4, while the same region from human nardilysin binds 1 mol of spermine/mol. Spermine enhanced the sensitivity of the mouse acidic domain, but not the human acidic domain, to protease V8. Together the data support a model where the acidic stretch of nardilysin functions as an autonomous domain. PMID- 11478916 TI - Expression of endothelial nitric oxide synthase in the ischemic penumbra: relationship to expression of neuronal nitric oxide synthase and vascular endothelial growth factor. AB - Expressional patterns of the endothelial and neuronal forms of nitric oxide synthase (NOS) in cerebral ischemia were studied utilizing a permanent middle cerebral artery occlusion (PMCAO) model. Motor performance and infarct volumes were determined in the rats. Immunohistochemical staining for eNOS, nNOS and neurofilament were performed at 1, 2, 3, 5, 7 and 14 days after PMCAO. Vascular endothelial growth factor (VEGF) expression was determined by in-situ hybridization. PMCAO caused a reproducible cortical infarct with motor deficits in the rats. Double immunohistochemical stainings indicated that eNOS and nNOS were induced in ischemic neurons. Most stained neurons were positive for both NOS forms but some reacted with only one NOS antibody. nNOS expression peaked at 24 48 h after PMCAO, stained mainly the cytoplasm of core neurons, and disappeared after the 3rd day. eNOS expression increased until the 7th day, stained mainly the cytoplasm and membrane of penumbral cells and disappeared by the 14th day after PMCAO. VEGF expression was significantly induced in the penumbral zone in a similar distribution to eNOS. The anatomical and temporal pattern of VEGF and eNOS induction in the brain after permanent ischemia suggest that these mediators may play a role in protecting penumbral tissue from additional ischemic damage. PMID- 11478917 TI - Discovery of a novel compound: insight into mechanisms for acrylamide-induced axonopathy and colchicine-induced apoptotic neuronal cell death. AB - The exposure of humans and experimental animals to certain industrial toxins such as acrylamide is known to cause nerve damage classified as axonopathy, but the mechanisms involved are poorly understood. Here we show that acrylamide induces morphological changes and tyrosine phosphorylation of focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (Pyk2), a member of the FAK subfamily, in human differentiating neuroblastoma SH-SY5Y cells. Furthermore, we identified a novel molecule designated 'compound-1' that inhibits the morphological and biochemical events. Daily oral administrations of the compound also effectively alleviated behavioral deficits in animals elicited by acrylamide in inclined plane testing, landing foot spread testing and rota-rod performance testing. The compound also effectively inhibited the biological and biochemical responses caused by another axonopathy inducer, colchicine, including tyrosine phosphorylation of Pyk2, formation of an 85-kDa poly(ADP-ribose)polymerase (PARP) fragment and apoptosis-associated induction of the NAPOR gene as well as neuronal cell death. Our findings not only provide insight into FAK and Pyk2 functions in neuronal cells, but may also be important in the development of therapeutic agents for peripheral neuropathy and neurodegeneration. PMID- 11478918 TI - Neuronal and inducible nitric oxide synthase expression and protein nitration in rat cerebellum after oxygen and glucose deprivation. AB - A perfusion model of global cerebral ischemia was used for the immunohistochemical study of changes in the glutamate-nitric oxide (NO) system in the rat cerebellum and cerebellar nuclei during a 0-14 h reperfusion period after 30 min of oxygen and glucose deprivation, with and without administration of 1.5 mM N(omega)-nitro-L-arginine methyl ester (L-NAME). While immunostaining for N methyl-D-aspartate receptor subunit 1 (NMDAR1) showed no marked changes during the reperfusion period, neuronal NO synthase (nNOS) immunostaining increased in stellate and basket cells, granule cells and neurons of the cerebellar nuclei. However, global cerebellar nNOS concentrations determined by Western blotting remained largely unchanged in comparison with actin expression. Inducible NOS (iNOS) immunostaining appeared in Purkinje cells and neurons of the cerebellar nuclei after 2-4 h of reperfusion and intensified during the 6-14 h period. This was reflected by an increase in global cerebellar iNOS expression determined by Western blotting. Immunostaining for protein nitrotyrosine was seen in Purkinje cells, stellate and basket cells, neurons of the cerebellar nuclei and glial cells in controls, and showed a progressive translocation in Purkinje cells and neurons of the cerebellar nuclei from an initial perinuclear or nuclear location towards the periphery. At the end of the reperfusion period the Purkinje cell apical dendrites were notably retracted and tortuous. Prior and concurrent L-NAME administration eliminated nitrotyrosine immunostaining in controls and blocked or reduced most of the postischemic changes observed. The results suggest that while nNOS expression may be modified in certain cells, iNOS is induced after a 2-4 h period, and that changes in protein nitration may be associated with changes in cell morphology. PMID- 11478919 TI - NXY-059, a novel free radical trapping compound, reduces cortical infarction after permanent focal cerebral ischemia in the rat. AB - Free radicals have gained wide acceptance as mediators of cerebral ischemic injury. It has previously been reported that a spin trap nitrone, alpha-phenyl-N tert-butyl nitrone (PBN), can reduce infarct volumes in rats subjected to either permanent or transient focal cerebral ischemia. A recent study has demonstrated that NXY-059, a novel free radical trapping nitrone compound, has a neuroprotective effect against transient focal cerebral ischemia. This study was designed to determine the effect of NXY-059 in a rodent model of permanent focal cerebral ischemia. Male spontaneously hypertensive rats were subjected to permanent middle cerebral artery occlusion (MCAO) by placement of a microaneurysm clip on the middle cerebral artery (MCA). Animals were divided into three groups: (1) physiological saline given as a 1 ml/kg i.v. bolus administered 5 min post MCAO followed immediately by a continuous i.v. infusion of 0.5 ml/h of physiological saline for 24 h (n=10); (2) 30 mg/kg, 1 ml/kg, i.v. bolus of NXY 059 dissolved in physiological saline administered 5 min post MCAO followed immediately by a continuous i.v. infusion of 30 mg/kg/h, 0.5 ml/h, of NXY-059 for 24 h (n=9); (3) 60 mg/kg, 1 ml/kg, i.v. bolus of NXY-059 dissolved in physiological saline administered 5 min post MCAO followed immediately by a continuous i.v. infusion of 60 mg/kg/h, 0.5 ml/h, of NXY-059 for 24 h (n=12). Infarction was quantified after a survival period of 24 h. Differences in infarct volume were examined with one-way ANOVA following Dunnet's multiple comparison test. The percentage of cortical infarction in the saline control group was 22.6 +/- 6.8% (mean+/-S.D.) of contra-lateral hemisphere, and in the 30 mg/kg/h NXY 059-treated group was 17.4% +/- 6.8% (NS). Plasma concentration (microM/l) of NXY 059 in the 30 mg/kg/h group was 80.2 +/- 52.2 (n=9), while in the 60 mg/kg/h group plasma concentration (microM/l) of NXY-059 was 391.0 +/- 207.0 (n=10). Infarction in the 60 mg/kg/h NXY-059-treated group was significantly reduced (P=0.009) to 14.5 +/- 5%. Our preliminary data demonstrate that administration of NXY-059 (60 mg/kg/h for 24 h) ameliorates cortical infarction in rats subjected to permanent focal cerebral ischemia with 24 h survival. PMID- 11478920 TI - NMDA receptor activation results in tyrosine phosphorylation of NMDA receptor subunit 2A(NR2A) and interaction of Pyk2 and Src with NR2A after transient cerebral ischemia and reperfusion. AB - Transient ischemia increases tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor. Several tyrosine kinases are involved in this process. In this study, effect of ischemia and reperfusion (I/R) on tyrosine phosphorylation of NMDA receptor subunit 2A (NR2A) and the interaction of two tyrosine kinases, Src and Pyk2, with NR2A was investigated. Four-vessel occlusion was used to produce transient (15 min) cerebral ischemia in SD rats. Tyrosine phosphorylation of NR2A in hippocampus was enhanced after 15 min of reperfusion and reached its peak level at 6 h of reperfusion. The increase sustained for at least 24 h. Src and Pyk2 co-immunoprecipitated with NR2A and the binding increased after I/R, which also reached a peak at 6 h of reperfusion. Besides, Src and Pyk2 were activated after I/R. These increases were prevented by ketamine, a selective NMDA receptor antagonist, which was administered to the SD rats 20 min before ischemia. Moreover, Src and Pyk2 coprecipitated with each other. These data show that NR2A, Src and Pyk2 might form a protein complex in vivo and the interaction suggests a possible mechanism of signal transduction in the postischemic hippocampus. PMID- 11478921 TI - Characterization of vinpocetine effects on DA and DOPAC release in striatal isolated nerve endings. AB - The effect of vinpocetine, a nootropic drug with anti-ischemic potential, on the release of DA and its main metabolite, DOPAC, was investigated in striatum isolated nerve endings under resting and depolarized conditions. Vinpocetine does not modify the baseline release of DA or the exocytotic release of DA evoked by high K(+), but inhibits the release of DA evoked by veratridine reversal of the DA transporter. In addition to these results, which confirm the vinpocetine selective blockade of voltage-sensitive presynaptic Na(+) channels (VSSC) previously reported [Neurochem. Res. 24 (1999) 1585], vinpocetine increases DOPAC release either under resting, veratridine or high K(+) depolarized conditions. This latter effect, which does not involve VSSC, was characterized. The parallel determination of the released and retained catecholamine concentrations revealed that vinpocetine increases DOPAC release at the expense of internal DA in a dose dependent manner (low microM range). In contrast to vinpocetine, the selective MAO-A inhibitor, clorgyline, increases DA and decreases DOPAC formation. The combined action of vinpocetine and clorgyline does not indicate, however, that the activation of MAO is the mechanism responsible for the increase in DOPAC caused by vinpocetine. Reserpine, although more potent and efficient than vinpocetine, qualitatively exerts the same pattern of changes on DA and DOPAC concentrations. It is concluded that, in addition to the inhibition of presynaptic VSSC permeability, which selectively inhibits the transporter mediated release of all neurotransmitters, vinpocetine increases DOPAC by impairing the vesicular storage of DA. Our results indicate that the cytoplasm extravesicular DA is metabolized by MAO to DOPAC. Most of the DOPAC formed is exported to the extracellular medium. PMID- 11478922 TI - Inhibition of prostaglandin synthesis reduces hyperthermic reactions induced by hypocretin-1/orexin A. AB - This experiment tested (i) the effect of orexin A injected into a lateral cerebral ventricle on sympathetic and thermogenic activity and (ii) the involvement of prostaglandins in these phenomena. The firing rates of the sympathetic nerves to interscapular brown adipose tissue (IBAT), along with IBAT and colonic temperatures and heart rate were monitored in urethane-anesthetized male Sprague-Dawley rats before and 6 h after an injection of orexin A (1.5 nmol) into the lateral cerebral ventricle. The same variables were monitored in rats with an intraperitoneal administration of lysine acetylsalicylate (100 mg/kg bw), an inhibitor of prostaglandins synthesis. The results show that orexin A increases the sympathetic firing rate, IBAT and colonic temperatures and heart rate. This increase is reduced by lysine acetylsalicylate. These findings suggest that orexin A affects sympathetic activity, which controls body temperature. Prostaglandins are involved in this control. PMID- 11478923 TI - The kappa-opioid antagonist GNTI reduces U50,488-, DAMGO-, and deprivation induced feeding, but not butorphanol- and neuropeptide Y-induced feeding in rats. AB - Antagonists selective for either kappa- [e.g. nor-binaltorphimine (nor-BNI)] and mu- (e.g. beta-funaltrexamine) opioid receptors have previously been shown to reduce both kappa- and mu-opioid-induced feeding. In the present studies, the anorectic effects of GNTI, a newly synthesized antagonist selective for kappa opioid receptors, were studied in rats. GNTI (0.032-0.32 nmol; i.c.v.), administered 15 min prior to food access, reduced feeding induced by the kappa opioid agonist U50,488 (producing a 70% maximal decrease), the mu-opioid agonist DAMGO (90% maximal decrease), and 24 h acute food deprivation (60% maximal decrease). GNTI did not reduce the orexigenic effects of butorphanol, an agonist that binds to both kappa- and mu-opioid receptors, and neuropeptide Y (NPY). Taken together, these results suggest that GNTI is a potent anorectic agent and opioid antagonist in rats. Like nor-BNI, GNTI reduced feeding induced by both kappa- and mu-opioid agonists. However, unlike nor-BNI, GNTI did not alter the orexigenic effects of butorphanol or NPY. Given the selectivity of GNTI and its effectiveness in several of the present experiments, its potency, and its short duration of action compared to nor-BNI, GNTI may serve to be a useful tool to study behavioral effects mediated by kappa-opioid receptors. PMID- 11478924 TI - Sleep deprivation decreases phase-shift responses of circadian rhythms to light in the mouse: role of serotonergic and metabolic signals. AB - The circadian pacemaker in the suprachiasmatic nuclei is primarily synchronized to the daily light-dark cycle. The phase-shifting and synchronizing effects of light can be modulated by non-photic factors, such as behavioral, metabolic or serotonergic cues. The present experiments examine the effects of sleep deprivation on the response of the circadian pacemaker to light and test the possible involvement of serotonergic and/or metabolic cues in mediating the effects of sleep deprivation. Photic phase-shifting of the locomotor activity rhythm was analyzed in mice transferred from a light-dark cycle to constant darkness, and sleep-deprived for 8 h from Zeitgeber Time 6 to Zeitgeber Time 14. Phase-delays in response to a 10-min light pulse at Zeitgeber Time 14 were reduced by 30% in sleep-deprived mice compared to control mice, while sleep deprivation without light exposure induced no significant phase-shifts. Stimulation of serotonin neurotransmission by fluoxetine (10 mg/kg), a serotonin reuptake inhibitor that decreases light-induced phase-delays in non-deprived mice, did not further reduce light-induced phase-delays in sleep-deprived mice. Impairment of serotonin neurotransmission with p-chloroamphetamine (three injections of 10 mg/kg), which did not increase light-induced phase-delays in non deprived mice significantly, partially normalized light-induced phase-delays in sleep-deprived mice. Injections of glucose increased light-induced phase-delays in control and sleep-deprived mice. Chemical damage of the ventromedial hypothalamus by gold-thioglucose (600 mg/kg) prevented the reduction of light induced phase-delays in sleep-deprived mice, without altering phase-delays in control mice. Taken together, the present results indicate that sleep deprivation can reduce the light-induced phase-shifts of the mouse suprachiasmatic pacemaker, due to serotonergic and metabolic changes associated with the loss of sleep. PMID- 11478925 TI - Activated astrocytes with glycogen accumulation in ischemic penumbra during the early stage of brain infarction: immunohistochemical and electron microscopic studies. AB - Brain infarction was induced in rats by injection of microspheres through the right internal carotid artery, and structural changes in the astrocytes were observed during the early period following the infarction. Necrotic foci, varying in size and shape, were found in the right hemisphere. After immunohistochemical staining for GFAP, GFAP-positive astrocytes in the perinecrotic area known as the ischemic penumbra had distinctly increased in number and size with elongation of cytoplasmic processes 3 days after infarction. Electron microscopic observation revealed that glycogen granules had markedly accumulated in the cytoplasm of astrocytes located in the ischemic penumbra 3 and 5 days after infarction. Seven days after infarction, however, the glycogen granules disappeared from the astrocytes. Intermediate filaments increasingly appeared in the protoplasmic astrocytes after 3 days and were abundant in the activated and hypertrophied astrocytes after 7 days. As a result of our present study, we conclude that: (1) the function of glucose uptake from blood vessels was not impaired in the astrocytes under hypoxic conditions; (2) the astrocytes actively ingested blood glucose through the endothelial cells and accumulated it as glycogen for activation of their functions, and the cell volume increased under hypoxic conditions; (3) the depression of energy metabolism and the decrease in the uptake of energy sources in the nerve cells promoted glycogen accumulation in the astrocytes under hypoxic conditions; (4) intermediate filaments (GFAP filaments) increased in number, coincident with the activation and enlargement of the astrocytes; and (5) protoplasmic astrocytes were transformed into fibrous astrocytes in the ischemic penumbra of the brain infarction. PMID- 11478926 TI - Opposite regulation of body temperature by cholinergic input to the paraventricular nucleus and supraoptic nucleus in rats. AB - Hypothalamic cholinergic system plays an important role in the regulation of body temperature and fluid balance. We have previously shown that cholinergic stimulation of the anterior hypothalamus and preoptic area was accompanied by a fall in body temperature, increased water intake, and increased Fos protein in the paraventricular nucleus (PVN) and supraoptic nucleus (SON). In the present study, to estimate the role played by cholinergic input to the PVN and SON in thermoregulation and water intake, we used microdialysis for cholinergic stimulation with neostigmine and analysis of the nucleus, and also investigated immunoreactivity for c-Fos protein in the brain. This stimulation increased extracellular concentration of acetylcholine in these nuclei. Stimulation of the PVN decreased body temperature and increased water intake. On the other hand, stimulation of the SON increased body temperature. Both in PVN-stimulated and SON stimulated rats, c-Fos-like immunoreactivity (Fos-IR) was evident in the PVN, SON and certain regions including locus coeruleus (LC), area postrema and nucleus of the solitary tract (NTS). Addition of atropine to the dialysis medium attenuated the increase of Fos-IR and suppressed the cholinergic stimulation-induced responses in body temperature and water intake. These results suggest that cholinergic muscarinic mechanisms in PVN and SON play an opposite function in the regulation of body temperature. The same neuronal pathway including LC and NTS may participate in an advance both in hypothermia and in hyperthermia. PMID- 11478927 TI - 5-HT(1B) and 5-HT(1D) receptors in the human trigeminal ganglion: co-localization with calcitonin gene-related peptide, substance P and nitric oxide synthase. AB - 5-Hydroxytryptamine (5-HT) is implicated in migraine and agonist directed against 5-HT(1B) and 5-HT(1D) receptors are commonly used as effective therapies. The antimigraine mechanisms involve the inhibition of intracranial sensory neuropeptide release. In order to determine which 5-HT(1) receptor subtypes are involved we have by immunocytochemistry examined the distribution of 5-HT(1B) and 5-HT(1D) receptors in the human trigeminal ganglia, and addressed which of them colocalize with calcitonin gene-related peptide (CGRP), substance P (SP) or nitric oxide synthase (NOS). We detected that 5-HT(1D) receptor immunoreactivity (i.r.) was predominantly expressed in medium-sized cells (86% of positive cells, 30-60 microm). About 9% of the 5-HT(1D) receptor i.r. cells were large in size (> 60 microm) and 5% were small in size (< 30 microm). In a similar pattern, 5 HT(1B) receptor i.r. was mainly expressed in medium-sized cells (81% in 30-60 microm, 15% in > 60 microm and 4% in < 30 microm). Double immunostaining was used to determine whether the 5-HT(1B) or 5-HT(1D) receptor immunoreactive cells co localized with either CGRP, SP or NOS. Thus, 89% of the CGRP i.r. cells expressed 5-HT(1D) receptor i.r. and 65% of the CGRP positive cells were 5-HT(1B) receptor positive. Most of the 5-HT(1D) (95%) and the 5-HT(1B) (94%) receptor i.r. cells showed SP immunostaining and 83% of 5-HT(1D) receptor and 86% of 5-HT(1B) receptor i.r. cells contained NOS. In conclusion, both 5-HT(1B) and 5-HT(1D) receptors are expressed in the human trigeminal ganglion and they are mainly localized in medium-sized cells and they seem to colocalize with CGRP, SP and NOS. PMID- 11478928 TI - Endogenous cannabinoid, 2-arachidonoylglycerol, attenuates naloxone-precipitated withdrawal signs in morphine-dependent mice. AB - In the present study, we examined the effects of endogenous ligand 2 arachidonoylglycerol (2-AG) on naloxone-precipitated withdrawal in morphine dependent mice, in comparison with that of two cannabinoid agonists, an ingredient of Cannabis sativa Delta(8)-tetrahydrocannabinol (Delta(8)-THC) and the synthetic cannabinoid CB1 receptor agonist HU-210. 2-AG at a dose of 10 microg per mouse (i.c.v.) significantly inhibited both jumping and forepaw tremor as signs of withdrawal following naloxone challenge in morphine-dependent mice. Furthermore, both Delta(8)-THC and HU-210 significantly attenuated these symptoms of withdrawal in morphine-dependent mice. Therefore, it is suggested that inactivation of the endogenous cannabinoid system is related to the induction of withdrawal syndrome in morphine-dependent mice. Moreover, hyperlocomotor activity in morphine-dependent mice was markedly increased by Delta(8)-THC 10 mg/kg, which had no effect in naive mice. This finding suggested that in morphine dependence, upregulation of cannabinoid CB1 receptors occurred. Non-psychoactive CB1 receptor agonists or accelerators of endocannabinoid synthesis may be potential as therapeutic drugs for opiate withdrawal symptoms. PMID- 11478929 TI - Acute melatonin and para-chloroamphetamine interactions on pineal, brain and serum serotonin levels as well as stress hormone levels. AB - para-Chloroamphetamine, an amphetamine analog, alters serotonergic neurochemistry. In previous reports, melatonin (MEL), when administered with other amphetamine analogs, altered the decline in serotonin content produced by these analogs. The present studies assessed the effects of various doses of melatonin and p-chloroamphetamine on serotonin levels in numerous brain regions in male rats. Melatonin (10, 25 or 50 mg/kg, s.c.) and p-chloroamphetamine (3 or 5 mg/kg, s.c.) were administered and, 3 h later, brain samples and serum were collected. Serotonin levels in the serum and various regions of the brain were assayed using high-performance liquid chromatography. Melatonin in combination with a high dose of p-chloroamphetamine (5 mg/kg) produced cumulative deficits in serotonin levels in the serum. However, serotonin levels in the pineal, cortex or brain stem in all combined melatonin and p-chloroamphetamine groups were not significantly different from groups that received p-chloroamphetamine alone. Serum adrenocorticotropin (ACTH) and corticosterone levels were significantly elevated in the melatonin and p-chloroamphetamine combined groups, suggesting that animals receiving both treatments were more stressed than control animals or animals receiving melatonin or p-chloroamphetamine alone. These results indicate that melatonin does not alter p-chloroamphetamine-induced deficits in central serotonin levels. The increased serum adrenocorticotropic hormone, corticosterone and serotonin levels observed following melatonin and p-chloroamphetamine treatment suggest that this combination may have adverse peripheral effects. PMID- 11478930 TI - Spinal N-acetyl-alpha-linked acidic dipeptidase (NAALADase) inhibition attenuates mechanical allodynia induced by paw carrageenan injection in the rat. AB - N-Acetylated-alpha-linked acidic dipeptidase (NAALADase) hydrolyzes N-acetyl aspartyl-glutamate (NAAG) to liberate N-acetyl-aspartate and glutamate. NAAG is a putative neurotransmitter and acts as a mixed agonist/antagonist on N-methyl-D aspartate (NMDA) receptors and acts as an agonist on the metabotropic glutamate receptor 3 (mGluR3). In the present study, we examined the role of spinal NAALADase in the maintenance of mechanical allodynia induced by carrageenan injection, skin incision and mild thermal injury using 2 (phosphonomethyl)pentanedioic acid (2-PMPA), a specific NAALADase inhibitor, in rats. Mechanical allodynia was induced by injection of 2 mg carrageenan into the paw (carrageenan model), by creating a 1-cm longitudinal skin incision of the plantar aspect of the foot (post-operative model), or by application of thermal stimulation (52.5 degrees C) for 45 s to the hind paw (mild thermal injury model). 2-PMPA was administered intrathecally at the time when the maximum mechanical allodynia occurred. Mechanical allodynia was assessed by the measurement of mechanical threshold using von Frey filaments. The mechanical threshold was measured 5, 15, 30, 60 and 90 min after the drug administration. In the carrageenan model, 100 microg of 2-PMPA attenuated the level of mechanical allodynia. 2-PMPA had no effect on the level of mechanical allodynia in both the post-operative pain model and the mild thermal injury model. These data suggested that the inhibition of spinal NAALADase alleviated mechanical allodynia induced by paw carrageenan injection. PMID- 11478931 TI - Calretinin and calbindin D-28k delay the onset of cell death after excitotoxic stimulation in transfected P19 cells. AB - In some neurological diseases, injury to neurones reflects an over-stimulation of their receptors for excitatory amino acids. This response may disturb the Ca(2+) homeostasis and lead to a pronounced and sustained increase in the intracellular concentration of this ion. On the basis of data derived from correlative studies, calcium-binding proteins have been postulated to play a protective role in these pathologies. We tested, directly, the capacity of the three calcium-binding proteins calretinin (CR), calbindin D-28k (CB) and parvalbumin (PV) to buffer [Ca(2+)], and to protect cells against excitotoxic death. We used P19 murine embryonic carcinoma cells, which can be specifically induced (by retinoic acid) to transform into nerve-like ones. The differentiated cells express functional glutamate-receptors and are susceptible to excitotoxic shock. Undifferentiated P19-cells were stably transfected with the cDNA for CR, CB or PV, induced to differentiate, and then exposed to NMDA, a glutamate-receptor agonist. The survival rates of clones expressing CR, CB or PV were compared with those of untransfected P19-cells using the lactate-dehydrogenase assay. CR- and CB expressing cells were protected from death during the first 2 h of exposure to NMDA. This protection was, however, transient, and did not suffice to rescue P19 cells after prolonged stimulation. Two of the three PV-transfected clones raised were vulnerable to NMDA-induced excitotoxicity; the third, which expressed the lowest level of PV, was protected to a similar degree as that found for the CR- and CB-transfected clones. Our results indicate that in the P19-cell model, CR and CB can help to delay the onset of cell death after excitotoxic stimulation. PMID- 11478932 TI - Presenilin-1 and its N-terminal and C-terminal fragments are transported in the sciatic nerve of rat. AB - The axonal transport of presenilin-1 was investigated in a spinal cord-sciatic nerve-neuromuscular junction model system in the rat. The technique of unilateral sciatic nerve ligation, using double ligatures, was combined with immunohistochemical staining and Western blotting to examine the axonal transport of the protein. Immunohistochemical studies involving the use of polyclonal antibodies for either the N-terminal or the C-terminal domain of presenilin-1 furnished evidence that both fragments may be present not only in the neuronal cell bodies, but also in the motoric and sensory axons and the motoric axon terminals at the neuromuscular junctions. After double ligation of the sciatic nerve for 6, 12 or 24 h, progressive immunostaining of presenilin-1 occurred above the upper ligature and to a lesser extent below the lower ligature. Double staining of the sciatic nerve for presenilin-1 and for amyloid precursor protein revealed overlapping immunoreactivity. Western blotting confirmed the accumulation of the approximately 20-kDa C-terminal and approximately 25-kDa N terminal fragments and the full-length 45-kDa holoprotein of presenilin-1 both above and below the ligature. It is concluded that besides the larger amounts of C-terminal and N-terminal fragments, a smaller quantity of intact presenilin-1 may be present and conveyed bidirectionally in the sciatic nerve of the rat. These results lend further support to the suggestion that presenilin-1 may leave the trans-Golgi network and be found in the axons and axon terminals of the various neurons. PMID- 11478933 TI - Sildenafil-induced peripheral analgesia and activation of the nitric oxide-cyclic GMP pathway. AB - Although several lines of evidence have shown a role of the nitric oxide/cyclic guanosine monophosphate signaling pathway in the nociceptive mechanism, the exact role of the phosphodiesterase (PDE) 5 enzyme via the NO-cGMP pathway is not fully understood in pain response. The present study was aimed at exploring the role of the NO-cGMP pathway in nociceptive conditions in experimental animals. Peripheral nociception was assessed by acetic acid-induced chemonociception or carrageenan induced hyperalgesia and central nociception was assessed by tail-flick and hot plate methods. Sildenafil exhibited dose-dependent (1, 2, 5 and 10 mg/kg, i.p.) antinociception in both male and female mice against acetic acid-induced writhing. However, it did not alter the pain threshold in central nociception (5 and 10 mg/kg, i.p.). Local administration of sildenafil (50-200 microg/paw, i.pl) also attenuated carrageenan-induced hyperalgesia. In the peripheral nociceptive reaction (acetic acid-induced chemonociception), the antinociceptive effect of sildenafil (2 mg/kg, i.p.) was enhanced by co-administration of sodium nitroprusside (0.25 mg/kg), and L-arginine (50 mg/kg). Sildenafil-induced analgesia was significantly blocked by methylene blue (1 mg/kg), a guanylate cyclase inhibitor, but was not reversed by L-NAME (10 mg/kg), a nitric oxide synthase inhibitor. But a higher dose of L-NAME (20 mg/kg) significantly reversed sildenafil analgesia. Both of these agents also reversed the facilitatory effect of L-arginine (50 mg/kg) and sodium nitroprusside (0.25 mg/kg) on sildenafil analgesia. These results suggest that sildenafil-induced analgesia is mediated via the inhibition of PDE5. The results also indicate that the guanylate cyclase system is stimulated in the peripheral nociceptive reaction. In conclusion, sildenafil produces antinociception and its effect can be potentiated by sodium nitroprusside and L-arginine, probably through the activation of the NO-cyclic GMP pathway. PMID- 11478934 TI - A dopaminergic mechanism is involved in the 'anxiogenic-like' response induced by chronic amphetamine treatment: a behavioral and neurochemical study. AB - The purpose of this study was to examine the influence of chronic d-amphetamine (AMPH) treatment (2 mg/kg i.p., for 9 consecutive days) on behavioral and neurochemical responses to a subsequent exposure - 4 days after the last AMPH injection--to the elevated plus-maze (EPM), as well as to determine the involvement of a dopaminergic mechanism in that influence. Results showed that chronic AMPH treatment induced an 'anxiogenic-like' response when animals were evaluated in the EPM test. Pretreatment with either haloperidol (HAL, 1 mg/kg i.p., 20 min prior to each injection) or SCH-23390 (0.1 mg/kg i.p., 10 min prior to each injection) completely abolished the chronic AMPH-induced 'anxiogenic like' effect displayed in the EPM test. However, sulpiride pretreatment (60 mg/kg i.p., 10 min prior to each AMPH injection) did not modify such effect. In addition, rats treated with AMPH and subsequently exposed to the EPM, showed a decrease in the maximal GABA-stimulated chloride uptake in cortical microsacs. HAL pretreatment restored the maximal chloride uptake induced by chronic AMPH. Altogether, these results suggest that: (1) previous exposure to chronic AMPH treatment induces an increased emotional response following a conflict situation, (2) dopamine D(1) receptors are mainly involved in chronic AMPH-induced changes in the behavior displayed in EPM test, and (3) an interaction between GABAergic and dopaminergic mechanisms may be implicated in neurochemical and behavioral changes induced by chronic AMPH treatment. PMID- 11478935 TI - Minocycline inhibits microglial activation and protects nigral cells after 6 hydroxydopamine injection into mouse striatum. AB - To determine the role of immune/inflammatory factors in dopaminergic cell degeneration in parkinsonian substantia nigra, we assayed tyrosine hydroxylase (TH)-positive immunoreactive neuronal numbers with stereologic techniques and CD11b-positive immunoreactive microglial profiles following 6-hydroxydopamine (6 OHDA) injection into ipsilateral striatum of mice. We further investigated the effect of minocycline on the inhibition of microglial activation and subsequent protection of nigral cells. The relative number of microglial profiles in the substantia nigra (SN) ipsilateral to the injection increased from 31 to 32% 1-3 days after injection, and increased further to 55% by 7 days and 59% by 14 days, compared with the contralateral SN. These changes started prior to the decrease of TH immunoreactivity of 34% on day 7 and of 42% by day 14. In animals treated with minocycline, microglial activation was inhibited by 47%, and TH positive cells were protected by 21% at day 14 after 6-OHDA injection, compared with those parkinsonian animals without minocycline treatment. All these results suggest that microglial activation may be involved in the nigral cell degeneration in 6 OHDA induced parkinsonian mice. PMID- 11478936 TI - Region-specific transcriptional response to chronic nicotine in rat brain. AB - Even though nicotine has been shown to modulate mRNA expression of a variety of genes, a comprehensive high-throughput study of the effects of nicotine on the tissue-specific gene expression profiles has been lacking in the literature. In this study, cDNA microarrays containing 1117 genes and ESTs were used to assess the transcriptional response to chronic nicotine treatment in rat, based on four brain regions, i.e. prefrontal cortex (PFC), nucleus accumbens (NAs), ventral tegmental area (VTA), and amygdala (AMYG). On the basis of a non-parametric resampling method, an index (called jackknifed reliability index, JRI) was proposed, and employed to determine the inherent measurement error across multiple arrays used in this study. Upon removal of the outliers, the mean correlation coefficient between duplicate measurements increased to 0.978+/ 0.0035 from 0.941+/-0.045. Results from principal component analysis and pairwise correlations suggested that brain regions studied were highly similar in terms of their absolute expression levels, but exhibited divergent transcriptional responses to chronic nicotine administration. For example, PFC and NAs were significantly more similar to each other (r=0.7; P<10(-14)) than to either VTA or AMYG. Furthermore, we confirmed our microarray results for two representative genes, i.e. the weak inward rectifier K(+) channel (TWIK-1), and phosphate and tensin homolog (PTEN) by using real-time quantitative RT-PCR technique. Finally, a number of genes, involved in MAPK, phosphatidylinositol, and EGFR signaling pathways, were identified and proposed as possible targets in response to nicotine administration. PMID- 11478937 TI - TGFbeta1 in liver fibrosis: time to change paradigms? PMID- 11478938 TI - Transforming growth factor beta signal transduction in hepatic stellate cells via Smad2/3 phosphorylation, a pathway that is abrogated during in vitro progression to myofibroblasts. TGFbeta signal transduction during transdifferentiation of hepatic stellate cells. AB - To current knowledge, transforming growth factor beta (TGFbeta) signaling is mandatory to establish liver fibrosis and various molecular interventions designed to affect the TGFbeta system were successfully used to inhibit fibrogenesis. Activated hepatic stellate cells (HSC), which are one important source of TGFbeta, are the major producers of extracellular matrix proteins in liver injury. We have previously shown that the TGFbeta response of this cell type is modulated during the transdifferentiation process. This work delineates the activation of TGFbeta downstream mediators, the Smads, in quiescent HSC and transdifferentiated myofibroblasts (MFB). The expression level of all Smads remained largely unchanged during this process. The response of HSC to TGFbeta, leading to, e.g., induction of alpha2 (I) collagen expression, is mediated by phosphorylation of Smad2 and Smad3 and subsequent nuclear translocation of a Smad containing complex. Neither TGFbeta-dependent nor endogenously phosphorylated Smad2/3 was detectable in comparable amounts in transdifferentiated MFB, indicating loss of TGFbeta sensitivity. Ectopic expression of Smad7 in HSC led to inhibition of Smad2 phosphorylation and abrogated TGFbeta response. In transdifferentiated MFB, expression of a constitutively active TGFbeta receptor I, but not treatment with TGFbeta1, resulted in transcriptional activation of a TGFbeta responsive promoter, thereby demonstrating completely restored TGFbeta signal transduction. Our data indicate that in contrast to a postulated mechanism of enduring autocrine TGFbeta signal transduction, early and late stages of HSC activation have to be distinguished, which is of importance for antifibrotic therapies. PMID- 11478939 TI - The importance of the C-terminal region and Cys residues for the membrane association of the NADPH:protochlorophyllide oxidoreductase in pea. AB - In vitro chloroplast import reactions and thylakoid association reactions have been performed with a series of C-terminal deletions and Cys-to-Ser substitution mutants of the pea NADPH:protochlorophyllide oxidoreductase (POR; EC 1.6.99). C terminal deletions of the precursor POR (Delta362-400, Delta338-400, Delta315-400 and Delta300-400) were efficiently translocated across the chloroplast envelope. However, except the Delta396-400 mutant, no C-terminal deletion mutants or Cys-to Ser substitution (Cys119, Cys281 and Cys309) mutants resisted post-treatment with thermolysin after the thylakoid association reactions. This suggests that these mutants were unable to properly associate to the thylakoids due to changes of the protein conformation of POR. PMID- 11478940 TI - Expression of human apolipoprotein A-I/C-III/A-IV gene cluster in mice reduces atherogenesis in response to a high fat-high cholesterol diet. AB - We have previously generated transgenic (Tg) mice expressing the human apolipoprotein (apo) A-I/C-III/A-IV gene cluster. This expression induced hyperlipidemia but reduced atherosclerotic lesions in genetically modified mice lacking apoE. Atherosclerosis is a multifactorial process and environmental factors such as diet play significant roles in its development. We examined here how an atherogenic diet influences the expression of the human genes and the characteristics of the Tg mice. Our results indicate that a high fat-high cholesterol diet up-regulates the intestinal expression of the three genes and the concentration of the three proteins in plasma. Cholesterol concentration was highly increased in the non-high density lipoprotein (HDL) fraction, and less, although significantly, in the HDL fraction. Tgs showed a 65% reduction in diet induced aortic lesions compared with non-Tg mice. Atherogenic diet increases the expression of the genes encoding the scavenger receptor class B type I (SR-BI) and ATP binding cassette transporter 1 (ABCA1) proteins. As cholesterol efflux mediated by SR-BI or by ABCA1 was enhanced in Tg mice fed an atherogenic diet, we can hypothesize that increased reverse cholesterol transport is the basis of the protective mechanism observed in these animals. In conclusion, we present evidence that the expression of the human gene cluster in mice protects against atherogenesis in response to an atherogenic diet. PMID- 11478941 TI - Effects of MAP kinase cascade inhibitors on the MKK5/ERK5 pathway. AB - Antibodies that recognise the active phosphorylated forms of mitogen-activated protein kinase (MAPK) kinase 5 (MKK5) and extracellular signal-regulated kinase 5 (ERK5) in untransfected cells have been exploited to show that the epidermal growth factor (EGF)-induced activation of MKK5 and ERK5 occurs subsequent to the activation of ERK1 and ERK2 in HeLa cells. The drugs U0126 and PD184352, which prevent the activation of MKK1 (and hence the activation of ERK1/ERK2), also prevent the activation of MKK5, although higher concentrations are required. Our studies define physiological targets of the MKK5/ERK5 pathway as proteins whose phosphorylation is largely prevented by 10 microM PD184352, but unaffected by 2 microM PD184352. Surprisingly, 2 microM PD184352 prolongs the activation of MKK5 and ERK5 induced by EGF or H(2)O(2), indicating negative control of the MKK5/ERK5 pathway by the classical MAPK cascade. Our results also indicate that ERK5 is not a significant activator of MAPK-activated protein kinase-1/RSK in HeLa cells. PMID- 11478942 TI - Identification of an expanded set of translationally active methionine analogues in Escherichia coli. AB - Amino acid incorporation into proteins in vivo is controlled most stringently by the aminoacyl-tRNA synthetases. Here we report the incorporation of several new methionine analogues into protein by increasing the rate of their activation by the methionyl-tRNA synthetase (MetRS) of Escherichia coli. cis-Crotylglycine (4), 2-aminoheptanoic acid (7), norvaline (8), 2-butynylglycine (11), and allylglycine (12) will each support protein synthesis in methionine-depleted cultures of E. coli when MetRS is overexpressed and the medium is supplemented with the analogue at millimolar concentrations. These investigations suggest important opportunities for protein engineering, as expansion of the translational apparatus toward other amino acid analogues by similar strategies should also be possible. PMID- 11478943 TI - Exon 3 of the alpha folate receptor gene contains a 5' splice site which confers enhanced ovarian carcinoma specific expression. AB - The human folate receptor (FR) is overexpressed in ovarian carcinoma. FR transcripts are heterogeneous due to the use of two promoters, P1 and P4, and alternative splicing of exon 3. RNase protection assay and RT-PCR revealed higher levels of the transcripts that include exon 3 in lines and specimens from ovarian carcinoma. A P1-chloramphenicol acetyltransferase (CAT) construct containing exon 3 demonstrated efficient reporter expression only in ovarian carcinoma. 5' and 3' deleted variants of the P1-CAT construct were analyzed by RT-PCR of the exogenous transcripts and reporter activity. A 5' splice site and 35 bp downstream intronic region of exon 3 appeared to regulate enhanced FR expression in ovarian carcinoma. PMID- 11478944 TI - In vitro selection of other proteins than antibodies by means of ribosome display. AB - The in vitro selection and simultaneous evolution of proteins is feasible by means of ribosome display. Here, we describe the use of a protein bearing a binding property without being an antibody for affinity enrichment of the ternary complex, consisting of a protein, a ribosome and a encoding mRNA. The binding property was a simple affinity tag, namely Strep-tag II and His-tag. We could demonstrate that the selection of a specific mRNA encoding a shortened bovine heart fatty acid binding protein with a N-terminal His-tag was possible. After nine cycles of transcription, translation, affinity selection and reverse transcription PCR the protein with the His-tag could be enriched 10(8)-fold. PMID- 11478945 TI - Nuclear localization signals in the Xenopus FGF embryonic early response 1 protein. AB - Xenopus early response 1 (XER1) is a fibroblast growth factor-inducible transcription factor whose developmentally regulated nuclear localization is thought to be important in the control of cell differentation during embryonic development [Luchman et al., Mech. Dev. 80 (1999) 111-114]. Analysis of the XER1 amino acid sequence revealed four regions which contain potential nuclear localization sequences (NLSs). Using mutant XER1 proteins and portions of XER1 fused to green fluorescent protein (GFP) transfected into NIH 3T3 cells, we have determined that only one of these, NLS4, located near the carboxy-terminus of XER1, is necessary and sufficient for targeting exclusively to the nucleus. Of the other three predicted NLS sequences, only NLS1, consisting of the sequence (138)RPRRCK(143) was shown to function as a cryptic, weak NLS. NLS4 contains a core region consisting of the sequence (463)RPIKRQRMD(471) which is similar to the core NLS directing the human c-MYC protein to the nucleus. The core sequence is flanked by a predicted cdc2/protein kinase A phosphorylation motif, however mutation of the serine(472) to alanine or aspartic acid had no detectable effect on accumulation of GFP-XER1 fusion proteins in the nucleus, demonstrating that this putative phosphorylation site plays no role in regulating nuclear transport. PMID- 11478946 TI - A novel grass pollen allergen mimotope identified by phage display peptide library inhibits allergen-human IgE antibody interaction. AB - The aim of this study was to investigate the molecular basis of human IgE allergen interaction by screening a phage-displayed peptide library with an allergen-specific human IgE-mimicking monoclonal antibody (mAb). A mAb that reacted with major grass pollen allergens was successfully identified and shown to inhibit human IgE-allergen interaction. Biopanning of a phage-displayed random peptide library with this mAb yielded a 12 amino acid long mimotope. A synthetic peptide based on this 12-mer mimotope inhibited mAb and human IgE binding to grass pollen extracts. Our results indicate that such synthetic peptide mimotopes of allergens have potential as novel therapeutic agents. PMID- 11478947 TI - Mice lacking histidine decarboxylase exhibit abnormal mast cells. AB - Histidine decarboxylase (HDC) synthesizes histamine from histidine in mammals. To evaluate the role of histamine, we generated HDC-deficient mice using a gene targeting method. The mice showed a histamine deficiency and lacked histamine synthesizing activity from histidine. These HDC-deficient mice are viable and fertile but exhibit a decrease in the numbers of mast cells while the remaining mast cells show an altered morphology and reduced granular content. The amounts of mast cell granular proteases were tremendously reduced. The HDC-deficient mice provide a unique and promising model for studying the role of histamine in a broad range of normal and disease processes. PMID- 11478948 TI - The cAMP response element binding protein-2 (CREB-2) can interact with the C/EBP homologous protein (CHOP). AB - cAMP response element binding protein-2 (CREB-2) is a basic leucine zipper (bZIP) factor that was originally described as a repressor of CRE-dependent transcription but that can also act as a transcriptional activator. Moreover, CREB-2 is able to function in association with the viral Tax protein as an activator of the human T-cell leukemia virus type I (HTLV-I) promoter. Here we show that CREB-2 is able to interact with C/EBP-homologous protein (CHOP), a bZIP transcription factor known to inhibit CAAT/enhancer-dependent transcription. Cotransfection of CHOP with CREB-2 results in decreased activation driven by the cellular CRE motif or the HTLV-I proximal Tax-responsive element, confirming that CREB-2 and CHOP can interact with each other in vivo. PMID- 11478949 TI - Direct production of an activated matrix metalloproteinase-9 (gelatinase B) from mammalian cells. AB - Matrix metalloproteinase-9 (MMP-9) is produced by the inactive proform and activated by a proteolytic process. However, it has not been reported to produce the active form directly from cells, which has hindered the research to elicit the physiological roles of this enzyme. In this study, we prepared mutant MMP-9 containing the furin-recognizing sequence in the prodomain and showed that the mutant MMP-9 was secreted as the active form directly from CHO-K1 cells and primary hepatocytes after the gene was transfected. The secreted MMP-9 showed proteolytic activity without further activation and degraded collagen IV in vitro. In addition, the transfection of the gene into the liver resulted in the efficient expression of active MMP-9 in the liver and the serum in vivo. PMID- 11478950 TI - Cholesterol effects on the phospholipid condensation and packing in the bilayer: a molecular simulation study. AB - A 15-ns molecular dynamics simulation of the fully hydrated liquid-crystalline dimyristoylphosphatidylcholine-cholesterol (DMPC-Chol) bilayer containing approximately 22 mol% Chol was carried out. The generated trajectory was analysed to investigate the mechanism of the Chol condensing effect on DMPC hydrocarbon chains and the influence of Chol on the chain packing in the membrane. Chol was found to induce stronger van der Waals interactions among the chains, whereas its interactions with the chains were weak. In the DMPC-Chol bilayer, as in the DMPC bilayer, DMPC chains were regularly packed around a chosen chain but around a Chol molecule they were not. DMPC gamma chains made closer contacts with Chol than the beta chains. PMID- 11478951 TI - Cone venom--from accidental stings to deliberate injection. AB - Cone snails have long been of note due to their colorful shells and deadly venom. Over the years, a number of people who have encountered these molluscs have been injured or killed by their sting. Biochemical analysis of the venom components has revealed a plethora of peptides and proteins that target a variety of receptors and ion channels. Pharmaceutical companies are now utilizing the selectivity and potency of Conus-derived peptides to develop novel medications for pain, epilepsy and other disorders. PMID- 11478952 TI - Production and specificity of monoclonal antibodies against nodularin conjugated through N-methyldehydrobutyrine. AB - Nodularin (Nod) is a cyclic pentapeptide hepatotoxin produced by the cyanobacterial genus Nodularia living in brackish waters and coastal lagoons. The toxicity of Nod is due to specific inhibition of the type-1 and type-2A intracellular protein phosphatases (PP1 and PP2A, respectively). We have developed a monoclonal antibody against Nod using chemical modification (aminoethylation) of one of its core amino acids, N-methyldehydrobutyrine. The developed antibody is highly specific for Nod, with negligible reactivity to the closely related cyanobacterial toxin microcystin (MC). The monoclonal antibody was employed for quantitative competitive ELISA assay. The analytical sensitivity of the assay was up to 0.2 ng/ml. Comparison of the developed ELISA test with HPLC-based measurements of Nod, with both laboratory and field samples, showed a good correspondence between the results yielded by these two methods. The antibodies developed by this technique provide means for developing extremely sensitive and specific analytical assays for direct measurement of nodularin and related toxins in cyanobacterial or water samples. PMID- 11478953 TI - Impact of a toxic and a non-toxic strain of Microcystis aeruginosa on the crayfish Procambarus clarkii. AB - The occurrence of cyanobacteria in many water bodies where crayfish such as Procambarus clarkii are abundant leads to the possibility of toxin accumulation and food chain transfer. This paper describes the accumulation and depuration of microcystins from a microcystin and a non-microcystin producing strain of Microcystis aeruginosa, on the survivorship, growth and nutritional status of P. clarkii. Crayfish larvae were resistant to cyanobacteria and their toxins, surviving cyanobacteria densities during acute exposures. Juvenile crayfish tolerated toxic cyanobacteria better than non-toxic ones. This effect was also observed when analysing nutritional status of crayfish fed toxic and non-toxic cyanobacteria with the former having better lipid and protein contents than those fed non-toxic Microcystis. P. clarkii accumulated up to 2.9 microg MCYST/dry crayfish weight and the depuration pattern was similar to that observed for mussels by other authors. Due to the fact that the major part of the toxin is accumulated in the intestine and in the hepatopancreas, there is no significant risk in terms of human health if these parts are removed prior to crayfish consumption. Nevertheless, their use in dairy food and the possible transference of toxins along food chains should not be disregarded. PMID- 11478954 TI - Effect of BJcuL (a lectin from the venom of the snake Bothrops jararacussu) on adhesion and growth of tumor and endothelial cells. AB - Lectins are polyvalent carbohydrate-binding proteins of non-immune origin. Recently, we have isolated and characterized a lectin from the venom of the snake Bothrops jararacussu. This lectin (BJcuL) has been shown to bind to lactose moieties and induce agglutination of erythrocytes. In the present work, we observed that cells from human metastatic breast cancer (MDA-MB-435) and human ovarian carcinoma (OVCAR-5) cell lines adhere, although weakly, to BJcuL. However, BJcuL did not inhibit adhesion of these cells to the extracellular matrix proteins fibronectin, laminin and type I collagen. Importantly, viability of these tumor cells and cells from other human tumor cell lines and a bovine brain endothelial cell line was suppressed by BJcuL. These findings suggest that the lectin BJcuL may serve as an interesting tool for combating tumor progression by inhibiting tumor cell and endothelial cell growth. PMID- 11478955 TI - Neutralization of the pharmacological effects of bothropstoxin-I from Bothrops jararacussu (jararacucu) venom by crotoxin antiserum and heparin. AB - Bothropstoxin-I (BthTX-I), the principal myotoxin of Bothrops jararacussu venom, is devoid of phospholipase A(2) (PLA(2)) activity but capable of blocking neuromuscular transmission in mouse nerve-muscle preparations. In this study, the ability of crotoxin antiserum and heparin in preventing the neurotoxic and myotoxic effects of BthTX-I was investigated. Phrenic nerve-diaphragm preparations (PND) stimulated indirectly with supramaximal stimuli (0.2 ms, 0.1 Hz) were incubated with BthTX-I (20 microg/ml) alone or with BthTX-I preincubated with antiserum or heparin for 30 min at 37 degrees C prior to testing. Control preparations were incubated with Tyrode solution, antiserum or heparin alone. BthTX-I (20 microg/ml) produced 50% neuromuscular blockade in the PND preparations in 31+/-4min, with complete blockade occurring in 120 min. The antiserum and heparin significantly prevented the neuromuscular blockade caused by BthTX-I (84 +/- 4% and 100% protection, respectively). Light microscopy examination of the muscles at the end of the 120 min incubation showed that BthTX I damaged 48 +/- 6% of the fibers. Preincubating the toxin with antivenom significantly reduced the extent of this damage (only 15 +/- 4% of fibers affected, corresponding to 69% protection, P<0.01) whereas heparin offered no protection (34 +/- 7% of fibers affected, not significantly different from that seen with toxin alone). These results show that the antivenom was more effective in neutralizing the myotoxic effects of BthTX-I than was heparin. PMID- 11478956 TI - Production of potent polyvalent antivenom against three elapid venoms using a low dose, low volume, multi-site immunization protocol. AB - The purpose of this study was to prepare a potent polyvalent antivenom against three elapids namely, the Thai cobra (Naja kaouthia, NK), the King cobra (Ophiophagus hannah, OH) and the banded krait (Bungarus fasciatus, BF). Two groups of horses were immunized. Group 1, comprising five horses, was immunized twice with a mixture of postsynaptic neurotoxins followed by an additional six immunizations with a mixture of crude venoms of the three elapids. Group 2, comprising four horses, was immunized with a mixture of crude venoms throughout the course. For the first immunization, the immunogens were emulsified in Complete Freund's adjuvant and injected using a low dose, low volume multi-site immunization protocol previously developed in this laboratory (Pratanaphon, R., Akesowan, S., Khow, O., Sriprapat, S. and Ratanabanangkoon, K. (1997) Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia). Vaccine 15, 1523-1528). The second immunization was carried out with the immunogens in Incomplete Freund's adjuvant. Blood was drawn to assay the antibody titer by ELISA. Sera at the peak of ELISA titers were pooled and assayed for the median effective dose (ED(50)). The ED(50)'s of antivenom from Group 1 horses against NK, OH and BF venoms were 1.44, 0.22 and 0.23 ml serum/mg venom, respectively, while those from Group 2 horse sera were 0.88, 0.20 and 0.49 ml serum/mg venom, respectively. The potency of sera from Group 2 against BF venom was significantly higher, while the potencies against NK and OH venoms were comparable to those of the corresponding monovalent antivenoms produced under the same protocol. This potent, truly polyvalent antivenom should be useful in saving lives of victims envenomed by these elapids and the immunization protocol should be useful in the production of potent polyvalent antivenoms against other medically important elapids. PMID- 11478957 TI - A hyaluronidase from Tityus serrulatus scorpion venom: isolation, characterization and inhibition by flavonoids. AB - The purification procedure of a hyaluronidase from Tityus serrulatus scorpion venom is described. It involves basically an ion-exchange chromatography on CM cellulose at pH 7.8 followed by a rechromatography of the active fraction on the same column at pH 4.7. The optima pH and temperature for maximum activity of the isolated enzyme was 6.0 and 40 degrees C, respectively. Its K(M) was 69.7 microg/ml at 37 degrees C and its specific activity was 19,900+/-1,730 turbidity reducing units (TRU)/mg against 845+/-88TRU/mg for the whole desiccated venom, representing a 23- to 24-fold purification range. The hyaluronidase activity of the purified protein (51kDa) was inhibited by some flavonoid compounds. This article also showed that T. serrulatus hyaluronidase affected on the activity of the venom's major toxin, tityustoxin-I (TsTX-I or Ts1), as reflected by alterations in the serum levels of creatine kinase (CK), lactate dehydrogenase (LD) and aspartate aminotransferase (AST) following injection of TsTX-I, in the presence or absence of hyaluronidase. PMID- 11478958 TI - Bothrops asper and Bothrops jararaca snake venoms trigger microbicidal functions of peritoneal leukocytes in vivo. AB - Venoms from snakes of the genus Bothrops cause pronounced local effects in the victims. These alterations result not only from the direct toxic action of venom components, but also from the prominent inflammatory reaction associated with these envenomations. In this study we investigated the ability of Bothrops asper (BaV) and Bothrops jararaca (BjV) venoms to induce cellular influx and microbicidal functions in leukocytes. BaV and BjV (5 microg/animal) caused a long lasting infiltration of leukocytes (3-48 h) when injected into mouse peritoneal cavity. Both venoms increased phagocytosis and production of hydrogen peroxide (H2O2) by polymorphonuclear (PMN) and mononuclear (MN) peritoneal leukocytes. In addition, nitric oxide (NO) production by macrophages was also enhanced after the venom injections. This effect was inhibited by treating animals with L-NAME and aminoguanidine, thus suggesting the induction of iNOS synthesis by the venoms. Western blot analysis confirmed the expression of iNOS in macrophages. BaV and BjV injection led to increased levels of IFN-gamma at the site of inflammation. Since IFN-gamma is an effective inducer of iNOS expression, an indirect action of the venoms on iNOS expression can be proposed. A marked formation of nitrotyrosine-containing proteins was also observed in macrophage homogenates. Based on these results, we suggest that reactive oxygen and nitrogen-derived species are involved in the pathogenesis of the local tissue damage characteristic of Bothrops sp envenomations. PMID- 11478959 TI - Botulinum neurotoxin types B and E: purification, limited proteolysis by endoproteinase Glu-C and pepsin, and comparison of their identified cleaved sites relative to the three-dimensional structure of type A neurotoxin. AB - Botulinum neurotoxin (NT) serotypes B and E are approximately 150 kDa proteins. Isolated from the liquid culture of Clostridium botulinum the NT type E is a single chain protein while the NT type B, from the proteolytic strain of the bacteria, is a mixture of dichain (nicked within a disulfide loop located about one-third the way from the N-terminus to the C-terminus) protein and its precursor single-chain protein. Endoproteinase Glu-C (EC 3.4.21.19) and pepsin (EC 3.4.23.1) were used for controlled digestion of NT types B and E; the amino acid residues flanking many of the cleavable peptide bonds were identified and the corresponding proteolytic fragments partially characterized. Chemical identification of 82 and 108 residues of types B and E NT, respectively, revealed that the residue 738 and 1098 in type E NT, identified as Leu and Asn, respectively, differ from those deduced from nucleotide sequences. Several fragments overlapped spanning various segments of the NT's functional domains; they appear to have potential for structure-function studies of the NT. The cleavage sites were compared with the previously determined proteolyzed sites on NT types A and E. The cleavage sites of the NT types A, B and E, all exposed on the protein surface, were scrutinized in the context of the three-dimensional structure of crystallized NT type A [Lacy, D.B., Stevens, R.C., 1999. J. Mol. Biol. 291, 1091-1104]. Detailed procedures for isolation of pure NT types B and E in large quantities (average yield 92 and 62 mg, respectively) suitable for crystallization are reported. PMID- 11478960 TI - Dynamics of C2 toxin and chlorophyll-a formation in the dinoflagellate Alexandrium tamarense during large scale cultivation. AB - The production of paralytic shellfish toxins (PSTs) by the dinoflagellate Alexandrium tamarense ATCI01, a toxigenic strain isolated from South China coastal waters, was studied in batch cultures in relatively large volumes (20l). Under nutrient-replete conditions, this strain produced C2 toxin (C2T) as a predominant PST. In a 15-day production culture, phosphate was depleted by day 4, the stationary phase began at day 6, and the toxin productivity peaked at day 10, in which the cell content of C2T reached 76 fmol per cell. Much of the toxin was produced after the depletion of phosphate in the medium suggesting that C2T is a secondary metabolite. Aeration with small bubbles was useful in increasing cell mass and toxin yield. Chlorophyll-a (Chl-a) was formed in algal cells until the culture entered the stationary phase, after which Chl-a began to disappear rapidly from the culture while the C2T content continued to rise. These results suggest a metabolic relationship between Chl-a and C2T. PMID- 11478961 TI - Toxin composition of the toxic dinoflagellate Prorocentrum lima isolated from different locations along the Galician coast (NW Spain). AB - The DSP toxin composition of 19 Prorocentrum lima isolates from different locations of the Galician rias (Vigo and Pontevedra) was investigated by high performance liquid chromatography coupled with fluorimetric detection. Boiling and freeze/thaw/hydrolyse methodology were applied during extraction to detect OA, DTX1, DTX2 and their esterified derivatives. OA and DTX2 were detected in both free and esterified form, the latter always in very low amounts, whilst DTX1 was always present in the free form. This indicate that the hypothesized self protection mechanism of toxin storage in the less active esterified forms does not seem to apply to DTX1. A slight increase in the toxin concentration per cell was found during growth, although toxin composition did not vary appreciably. Toxin production and toxin profile varied significantly depending on the isolate. Four groups of P. lima were differentiated by cluster analysis according to their toxin composition. It is noteworthy that one of the clusters comprised all the strains collected from one location characterised by its geographical isolation, whereas the other clusters consisted of isolates from different locations. The differences in the toxin profile from P. lima strains and from the DSP contaminated shellfish, together with the very good correlation between Dinophysis spp occurrence and DSP toxicity in shellfish, support that these planktonic species are the main agents responsible for DSP events in Galicia. PMID- 11478962 TI - Properties of St I and St II, two isotoxins isolated from Stichodactyla helianthus: a comparison. AB - Sticholysins I and II are two highly hemolytic polypeptides purified from the Caribbean Sea anemone Stichodactyla helianthus. Their high sequence homology (93%) indicates that they correspond to isoforms of the same hemolysin. The spectroscopic measurements show a close similarity in the secondary structure content, conformation and stability of both toxins. Exposure of the toxins to high pHs (>11), a free radical source (AAPH), urea or temperature produce permanent changes in the toxin that lead to a significant loss of HA. It is significant to note that this loss of hemolytic activity occurs when other indicators, probably with the only exception of near-UV CD spectra, barely detect changes in the protein structure. This emphasizes the sensitivity of the protein function to changes in the macromolecule conformation. The most noticeable difference between both toxins is the considerably higher activity of St II, both measured in terms of erythrocyte internal K(+) exit or hemolysis; which is related to enthalpic factors. This difference is not due to an incomplete association of St I to the membrane. We consider then that the different pore forming capacity of both toxins in erythrocytes can be explained in terms of the difference in charge of the N-terminal fragment, than can considerably reduce the St I insertion rate in the membrane probably due to the negatively charged outer leaflet of the red blood cell, without a significant reduction of its capacity to bind to the cell membrane. This electrostatic effect, together with a slightly more relaxed structure in St II, could explain the higher pore forming capacity of St II in the red blood cell membrane. PMID- 11478963 TI - Enhancing the hypotensive effect and diminishing the cytolytic activity of hornet mastoparan B by D-amino acid substitution. AB - Mastoparan B (MP-B) is a cationic tetradecapeptide (LKLKSIVSWAKKVL-CONH(2)) isolated from the venom of the Taiwan hornet Vespa basalis. Unlike other vespid mastoparans, the peptide is capable of inducing short-term hypotension and causes hemolysis in animals. This study was aimed to find out MP-B analogs that possess higher hypotensive potency with the least lytic action by D-amino acid substitution, especially at lysine (Lys) residues. The synthetic MP-B isomer in which Lys(2) was replaced by D-Lys showed a significant decrease in both hemolytic and hypotensive activities. Substitution of Lys(4) by D-Lys in MP-B also caused a marked reduction of hemolytic activity, but its hypotensive action was only slightly affected. However, when Lys(11,12) were replaced by D-Lys, the resulting isomer ([D-Lys(11,12)]MP-B) exhibited a higher hypotensive activity with negligible hemolytic activity as compared with the native peptide. The D antipot of MP-B in which all amino acid residues were replaced by D-isomers showed the highest hypotensive activity with a hemolytic activity about 1/5 that of MP-B. The results reveal that D-Lys substitution at the N-terminus of MP-B (Lys(2,4)) causes decreases in both hypotensive and hemolytic activities, while D Lys substitution at the C-terminus (Lys(11,12)) leads to a significant increase in hypotensive activity of MP-B with a remarkable decrease in hemolytic activity. The hypotensive effect of [D-Lys(11,12)]MP-B was more prominent on spontaneously hypertensive rats. At a proper dose (0.3mg/kg) the peptide could reduce the high blood pressure (approximately 180 mmHg) of the rat to a normal level (approximately 120 mmHg) for more than 3h. [D-Lys(11,12)]MP-B which possesses a potent hypotensive action with the least cytolytic side effect is the best MP-B analog for studying the mechanism of cardiovascular inhibition by MP-B and could be useful as a hypotensive agent in hypertension crisis. PMID- 11478964 TI - The effect of jararhagin, a metalloproteinase from Bothrops jararaca venom, on pro-inflammatory cytokines released by murine peritoneal adherent cells. AB - The release of pro-inflammatory cytokines (IL-1beta, IL-6 and TNF-alpha) from murine peritoneal adherent cells (MPAC) was studied after exposure to jararhagin, a metalloproteinase/disintegrin isolated from Bothrops jararaca venom. MPACs were treated with LPS (lipopolysaccharide), jararhagin, or EDTA-inactivated jararhagin for up to 24h. Following incubation, the culture supernatant was assayed by ELISA for the presence of cytokines, while the cells were analysed for viability and cytokine mRNA expression. The cells exposed to native jararhagin released TNF alpha and IL-1beta after 4 and 24h respectively. When MPACs were exposed to Jararhagin treated with EDTA, TNF-alpha and IL-1beta production was sustained throughout the culture period and IL-6 production was observed. TNF-alpha, IL-6 and IL-1beta mRNA were detected 4h after stimulation with either native or EDTA treated jararhagin. Addition of jararhagin to LPS stimulated cells resulted in a dramatic decrease in the release of IL-6 and TNF-alpha. RT-PCR showed that this inhibition does not occur at the transcriptional level and further experiments showed that jararhagin degraded soluble cytokines by proteolytic activity. This study suggests that jararhagin induces TNF-alpha, IL-1beta and IL-6 expression, which may be rapidly degraded by its proteolytic activity. PMID- 11478965 TI - Decomposition of microcystin-LR by Fenton oxidation. AB - A novel and promising method of microcystin-LR (mcyst-LR) degradation is reported. The decomposition of this cyanobacterial toxin using Fenton reagent has been observed with very low initial concentrations of H2O2 and Fe2+ (Fe3+) in the reaction mixture. Mcyst-LR was isolated from a laboratory culture of Microcystis aeruginosa PCC 7813. The initial concentration of the toxin used exceeded by several orders of magnitude those occurring naturally in lakes and drinking water. Even so, the decomposition of the toxin was complete after 30 min. PMID- 11478966 TI - Characterisation of the gene encoding the alpha-toxin Amm V from the scorpion Androctonus mauretanicus mauretanicus. AB - The full-length cDNA encoding the scorpion alpha-toxin Amm V was amplified from a cDNA library produced from the venom glands of the scorpion Androctonus mauretanicus mauretanicus from Morocco. We deduced the amino acid sequence of the encoded precursor protein and found that the mature toxin was similar to the previously characterised toxin. The genomic DNA sequence encoding the toxin was also amplified, subcloned and sequenced. This also led to the isolation of a new Amm V related-gene. Then, for the first time, we studied changes in the level of toxin mRNA synthesis over time. PMID- 11478967 TI - Identification of the myotoxic site of the Lys49 phospholipase A(2) from Agkistrodon piscivorus piscivorus snake venom: synthetic C-terminal peptides from Lys49, but not from Asp49 myotoxins, exert membrane-damaging activities. AB - Group II phospholipase A(2) (PLA(2)) myotoxins found in the venoms of Crotalidae snakes can be divided into 'Asp49' and 'Lys49' isoforms, the latter being considered catalytically-inactive variants. Previous studies on one Lys49 isoform, myotoxin II from Bothrops asper, indicated that its myotoxic activity is due to the presence of a short cationic/hydrophobic sequence (115-129) near its C terminus, which displays membrane-damaging properties. Since the C-terminal region of different group II PLA(2) myotoxins presents considerable sequence variability, synthetic peptides homologous to region 115-129 of myotoxin II, but corresponding to B. asper myotoxin III (Asp49), Agkistrodon piscivorus piscivorus Asp49 PLA(2) and Lys49 PLA(2), were studied to determine the possible functional relevance of such region for the toxic activities of these proteins. Results showed that both Lys49-derived peptides (p-BaK49 and p-AppK49) were able to lyse skeletal muscle C2C12 cells in culture, and to induce edema in the mouse footpad assay. Moreover, p-AppK49, which showed a markedly stronger cytotoxic potency than p-BaK49, additionally induced skeletal muscle necrosis when injected into mice. These observations unequivocally identify the sequence 115-129 (KKYKAYFKLKCKK) of the Lys49 PLA(2) of A. p. piscivorus as containing the key structural determinants needed for myotoxicity, and represent the first report of an unmodified, PLA(2)-derived short synthetic peptide with the ability to reproduce this effect of a parent toxin in vivo. On the other hand, the two Asp49 derived peptides did not show any toxic effects in vitro or in vivo, even at high concentrations. These findings suggests that Lys49 and Asp49 group II PLA(2)s might exert their myotoxic actions through different molecular mechanisms, by implying that the latter may not utilize their C-terminal regions as main membrane-destabilizing elements. PMID- 11478968 TI - Expression of an active recombinant lysine 49 phospholipase A(2) myotoxin as a fusion protein in bacteria. AB - ACL myotoxin (ACLMT) is a K49 phospholipase A(2)-like protein isolated from the venom of the snake Agkistrodon contortrix laticinctus (broad-banded copperhead) that induces necrosis of skeletal muscle. We have previously cloned and sequenced the cDNA coding for ACLMT from a venom gland cDNA library. In order to perform structure and function studies, we have developed an expression system for production of ACLMT as a fusion protein with maltose binding protein (MBP) from the periplasm of bacteria, using the pMAL-p2 expression vector. The cDNA coding for the mature toxin without the signal peptide was amplified by PCR and subcloned into the pMAL-p2 vector. The new plasmid (pMAL-MT) was used to transform BL21(DE3) E. coli cells. Culture of transformed cells induced with IPTG led to the expression of a 60 kDa fusion protein which strongly reacts with anti native ACLMT antibodies. The fusion protein was purified from the bacterial periplasm by affinity chromatography in an amylose column and by gel filtration. The purified fusion protein (MBP-rACLMT) was able to induce necrosis of skeletal muscle of mice very similar to that caused by the native myotoxin. PMID- 11478969 TI - Seasonal changes in the composition of the essential oil extract of East Mediterranean sage (Salvia libanotica) and its toxicity in mice. AB - Sage (Salvia libanotica) is an East Mediterranean plant, the extract of which is used for the treatment of colds, coughs, and stomach ache. Experimental studies on the toxicity of its oil are scarce despite its wide use in traditional medicine. This study aims to provide data on its acute toxicity and to investigate the relationship between seasonal changes in oil composition and toxicity. The composition of the oil extract from the leaves of this plant was determined at four different times of the year; August (summer), October (fall), January (winter) and April (spring). The toxicity of each fraction was investigated following intraperitoneal (i.p.) injection into mice. Distillations of oils from plants and GC analyses revealed that the main constituent of sage oil is 1,8-cineole. Other components included ketones such as camphor and alpha,beta-thujone, terpenes such as limonene and alpha,beta-pinene, and alcohols such as borneol and linalool. Major seasonal changes were found in the composition of the oil. Essential oil extracted from plants collected in the winter season (January) contained higher levels of camphor (12.3%), alpha,beta thujone (1.9%), and camphene (4.8%). The winter extract was found to be the most toxic, (LD(50): 839 mg/kg body weight) and exhibited powerful convulsant properties. This indicates a strong correlation between the contents of camphor, thujones and camphene and the oils' toxicity. The spring extract was the least toxic (LD(50): 1200 mg/kg body weight) and contained lower levels of camphor (7.7%), alpha,beta-thujone (1.3%) and camphene (3.1%). Thus, we recommend that oil extracts of sage marketed for use in certain unconventional medicines be prepared from spring plants. PMID- 11478970 TI - Determination of the neutralizing potency of horse antibothropic and anticrotalic antivenoms in blood samples collected on filter paper. AB - The correlation coefficients between in vivo neutralization of lethal toxicity (ED(50)) and levels of antibodies measured by enzyme-linked immunosorbent assay (ELISA) in blood samples collected on filter paper were investigated to test the potency of horse antibothropic and anticrotalic antivenoms. Sixteen horses were hyperimmunized with Bothrops venom (50% from B. jararaca and 12.5% each from B. alternatus, B. jararacussu, B. neuwiedii and B. moojeni) and 12 horses with Crotalus durissus terrificus venom. Crude venom of C. d. terrificus and the lethal fraction of B. jararaca venom were used as antigens to set up an indirect ELISA. The correlation coefficient between ED(50) and ELISA antibodies titers against C. d. terrificus and the lethal fraction of B. jararaca venom was r = 0.8 (P<0.001) and r = 0.78 (P<0.001), respectively. PMID- 11478971 TI - Genetic engineering of neural function in transgenic rodents: towards a comprehensive strategy? AB - As mammalian genome projects move towards completion, the attention of molecular neuroscientists is currently moving away from gene identification towards both cell-specific gene expression patterns (neuronal transcriptions) and protein expression/interactions (neuronal proteomics). In the long term, attention will increasingly be directed towards experimental interventions which are able to question neuronal function in a sophisticated manner that is cognisant of both transcriptomic and proteomic organization. Central to this effort will be the application of a new generation of transgenic approaches which are now evolving towards an appropriate level of molecular, temporal and spatial resolution. In this review, we summarize recent developments in transgenesis, and show how they have been applied in the principal model species for neuroscience, namely rats and mice. Current concepts of transgene design are also considered together with an overview of new genetically-encoded tools including both cellular indicators such as fluorescent activity reporters, and cellular regulators such as dominant negative signalling factors. Application of these tools in a whole animal context can be used to question both basic concepts of brain function, and also current concepts of underlying dysfuction in neurological diseases. PMID- 11478972 TI - Application of neurosonography to experimental physiology. AB - When Horsley and Clark invented the stereotaxic technique they revolutionized experimental neurobiology. For the first time it became possible to repeatably place experimental or surgical probes at precise locations within the skull. Unfortunately, variations in the position and size of neuroanatomical structures within the cranium have always limited the efficiency of this technology. Recent advances in diagnostic medical ultrasonography, however, allow for the real-time visualization of anatomical structures, in some cases with resolutions of up to 150 microm. We report here that commercially available ultrasonographs can be used in the laboratory to generate real-time in vivo images of brain structures in both anesthetized and awake-behaving animals. We found that ultrasonic imaging is compatible with many types of experimental probes including single neuron recording electrodes, microinjection pipettes, and electrodes for producing electrolytic lesions. Ultrasonic imaging can be used to place, monitor and visualize these probes in vivo. In our hands, commercially available ultrasonic probes designed for pediatric use allowed us to visualize anatomical structures with sub-millimeter resolution in primate brains. Finally, ultrasonic imaging allowed us to reduce the risk of accidentally damaging major blood vessels, greatly reducing the incidence of stroke as an unintended complication of an experimental neurosurgical procedure. Diagnostic ultrasound holds the promise of reducing the uncertainty associated with stereotaxic surgery, an improvement which would significantly improve the efficiency of many neurobiological investigations, reducing the number of animal subjects employed in this research. While this demonstration focuses on sonographic imaging in non-human primates, similar advances should also be possible for studies in other species, including rodents. PMID- 11478973 TI - Statistical aspects of quantitative image analysis of beta-amyloid in the APP(V717F) transgenic mouse model of Alzheimer's disease. AB - Cerebral beta-amyloidosis is a central part of the neuropathology of Alzheimer's disease (AD). Quantitation of beta-amyloid plaques in the human AD brain, and in animal models of AD, is an important study endpoint in AD research. Methodologic approaches to the measurement of beta-amyloid in the brain vary between investigators, and these differences affect outcome measures. Here, one quantitative approach to the measurement of beta-amyloid plaques in brain sections was analyzed for sources of variability due to sampling. Brain tissue was from homozygous APP(V717F) transgenic male mice. Sampling variables were at the mouse and microscopic slide and field levels. Results indicated that phenotypic variability in the mouse sample population was the largest contributor to the standard error of the analyses. Within each mouse, variability between slides or between fields within slides had smaller effects on the error of the analyses. Therefore, when designing studies of adequate power, in this and in other similar models of cerebral beta-amyloidosis, sufficient numbers of mice per group must be included in order for change in mean plaque burden attributable to an experimental variable to outweigh phenotypic variability. PMID- 11478974 TI - New strategies for embedding, orientation and sectioning of small brain specimens enable direct correlation to MR-images, brain atlases, or use of unbiased stereology. AB - We present a newly developed brain slicing machine and technique for tissue embedding, which enable orientation of fresh or fixed brain tissue from small laboratory animals, in any given position, and subsequent tissue sectioning into slabs with an optional thickness between 0.5 and 20 mm. The oriented tissue slabs may be analysed directly, or processed further on a cryostat or vibratome, into thin stainable histological sections, and aligned to MR-images or brain atlases, depending on the reference used for the initial orientation. Additionally, we describe a new embedding medium (HistOmer) which is an alginate cold polymer ready for instant use after mixing with water. HistOmer allows accurate positioning of the tissue during embedding, and at the same time supports and protects the embedded tissue during sectioning. HistOmer is, therefore, described comprehensively and compared with other commonly used embedding media. This novel slicing technique may also, as illustrated, be used to perform isotropic random orientation of the embedded tissue, before sectioning into tissue slabs of the same thickness. The technique thereby fulfills the requirements for optimal tissue sampling and preparation needed for an unbiased stereologic analysis. PMID- 11478975 TI - Simultaneous measurement of cerebral blood flow and mRNA signals: pixel-based inter-modality correlational analysis. AB - The analysis of pixel-based relationships between local cerebral blood flow (LCBF) and mRNA expression can reveal important insights into brain function. Traditionally, LCBF and in situ hybridization studies for genes of interest have been analyzed in separate series. To overcome this limitation and to increase the power of statistical analysis, this study focused on developing a double-label method to measure local cerebral blood flow (LCBF) and gene expressions simultaneously by means of a dual-autoradiography procedure. A 14C-iodoantipyrine autoradiographic LCBF study was first performed. Serial brain sections (12 in this study) were obtained at multiple coronal levels and were processed in the conventional manner to yield quantitative LCBF images. Two replicate sections at each bregma level were then used for in situ hybridization. To eliminate the 14C iodoantipyrine from these sections, a chloroform-washout procedure was first performed. The sections were then processed for in situ hybridization autoradiography for the probes of interest. This method was tested in Wistar rats subjected to 12 min of global forebrain ischemia by two-vessel occlusion plus hypotension, followed by 2 or 6 h of reperfusion (n=4-6 per group). LCBF and in situ hybridization images for heat shock protein 70 (HSP70) were generated for each rat, aligned by disparity analysis, and analyzed on a pixel-by-pixel basis. This method yielded detailed inter-modality correlation between LCBF and HSP70 mRNA expressions. The advantages of this method include reducing the number of experimental animals by one-half; and providing accurate pixel-based correlations between different modalities in the same animals, thus enabling paired statistical analyses. This method can be extended to permit correlation of LCBF with the expression of multiple genes of interest. PMID- 11478976 TI - High throughput screens for the identification of compounds that alter the accumulation of the Alzheimer's amyloid beta peptide (Abeta). AB - Evidence gathered over the last two decades suggests that beta amyloid (Abeta), the predominant proteinaceous component of senile plaques, plays an early and critical role in the etiology and pathogenesis of Alzheimer's disease (AD). Thus, it is reasonable to hypothesize that compounds capable of reducing the accumulation of Abeta may be of value therapeutically. Additionally, compounds that influence Abeta accumulation may be useful as tools to further dissect the cellular pathways that regulate Abeta production and accumulation. To screen for compounds that affect Abeta levels, we have established high throughput, cell based assays capable of the sensitive and selective detection of Abeta40 in parallel with the more amyloidogenic form of the peptide, Abeta42. To validate the approach, we examined the effects of several compounds previously identified to influence Abeta accumulation. Analysis of peptide accumulation following treatment with these compounds showed results similar to those previously published. Currently, we are using this assay to screen drugs that have already received FDA approval for the treatment of other diseases and over-the-counter natural product extracts. If compounds such as these can be identified that lower Abeta in the brain, they may represent one of the fastest and most cost effective methods to therapy. PMID- 11478977 TI - Validity of a quantitative technique to study striatal dopaminergic neurodegeneration by in vivo microdialysis. AB - The development of a technique that allows the direct quantitative study of the damage produced by a toxin on a specific neurotransmitter system is very important. For that, we have used the microdialysis technique to validate a method to study the specific drug's toxicity on dopaminergic (DAergic) striatal terminals. We perfused different MPP(+) and 6-hydroxydopamine (6-OHDA) concentrations, with different toxicity for DAergic terminals, 24 h after the implantation of the microdialysis probe (day 1). One day later (day 2), MPP(+) was perfused through the microdialysis probe and DA extracellular output measured. We hypothesize that the amount of extracellular dopamine (DA) obtained on day 2 is directly proportional to the neurotoxic damage produced on day 1. To corroborate this hypothesis tyrosine hydroxylase (TH) immunohistochemistry was also carried out on day 2. There was a clear correlation index between the amount of DA measured after MPP(+) perfusion and the lack of TH immunoreactivity measured as the radius of the area showing decrease in TH immunoreactivity around the cannula. These results show the possibility to measure DAergic remaining terminals after a toxic drug exposure by in vivo MPP(+) perfusion. The possibility to extend this neurotoxic study to another neurotransmitter systems is suggested. PMID- 11478978 TI - Cyclic AMP imaging in neurones in brain slice preparations. AB - The second messenger cascade of cyclic AMP (cAMP) plays an important physiological role in neurones, modulating neuronal excitability and synaptic transmission. The fluorescent probe FlCRhR allows real time ratiometric imaging of cAMP changes inside cells (Nature 349 (1991) 694). Until now, the only way to introduce FlCRhR into cells was microinjection, which restricted the use of FlCRhR to large invertebrate neurones. This report describes the use of the patch clamp technique to deliver FlCRhR into the cytosol of several types of neurones in brain slice preparations. Direct activation of adenylate cyclase by forskolin produced marked increases in fluorescence ratio, confirming that the probe can report cAMP increases. However, some neurones failed to exhibit a cAMP response and this lack of response was related to the nucleus integrity. Stimulation of membrane receptors positively coupled to adenylate cyclase elicited cAMP increases in various neuronal cell types. This is the first report of a cAMP response to neuromodulators measured by an imaging technique in neurones in brain slices. The method described here could find many applications such as testing the ability of agonists to specifically activate the cAMP cascade in identified neurones, studying the kinetics of the cAMP response and determining the subcellular localisation of cAMP changes. PMID- 11478979 TI - A novel mechanical dissociation technique for studying acutely isolated maturing Drosophila central neurons. AB - A novel mechanical method, for studying acutely isolated maturing Drosophila central neurons, has been developed. Electrophysiological experiments have been carried out to assess the quality of these acutely dissociated neurons. The mechanically dissociated Drosophila central neurons were divided into three categories depending on their size and morphological features. Four types of whole-cell K(+) currents were identified in these neurons, based on their kinetic properties. Moreover, the K(+) currents in the new preparation were found to have similar electrophysiological and pharmacological properties to those reported in the cultured neurons. The new technique, however, was more rapid and convenient. Furthermore, this new system was successfully applied to the isolation of neurons from the adult Drosophila, a process that is extremely difficult by routinely used methods. Thus, this new preparation would be very reliable and applicable to preparing Drosophila central neurons for biophysical and physiological studies. PMID- 11478981 TI - An elusive receptor is finally caught: P2Y(12'), an important drug target in platelets. AB - Despite intensive research, the nucleotide P2 receptor that is involved in the aggregation and activation of platelets by ADP has remained elusive. However, now two research groups have independently identified a new platelet receptor of unexpected structure, P2Y(12), that acts with the P2Y(1) receptor to form the site of ADP activation and explains the multiple transduction mechanisms observed in response to ADP in platelets. Recent evidence also suggests that a third component, ATP action on the P2X(1) receptor ion channel, contributes to platelet activation. PMID- 11478983 TI - Not much good is left when glia withdraw. PMID- 11478982 TI - Lipoxins: revelations on resolution. AB - Lipoxins (LXs) are endogenously produced eicosanoids typically generated during cell-cell interactions. In this article, the compelling evidence from in vitro and in vivo model systems that LXs actively promote the resolution of inflammation is reviewed. Of particular interest are indications that stable synthetic analogues of LXs and aspirin-triggered 15-epi-LXs can mimic many of the desirable anti-inflammatory, "pro-resolution" actions of native LXs. Given the enhanced stability and efficacy of these compounds a role as novel anti inflammatory therapeutics is proposed. PMID- 11478984 TI - A gene for Crohn's disease is given the nod. PMID- 11478985 TI - How can anti-diabetics suppress tumours? PMID- 11478986 TI - Tightening tobacco legislation. PMID- 11478987 TI - Good news for sick Kenyans. PMID- 11478988 TI - Microbots in the body. PMID- 11478989 TI - Feeling forgetful? PMID- 11478991 TI - Acupuncture and opioids. PMID- 11478992 TI - Crucial copper transporter. PMID- 11478993 TI - Monthly pill. PMID- 11478996 TI - Clinical trial safety. PMID- 11478994 TI - Competition for Viagra. PMID- 11478997 TI - Allison's trail of destruction. PMID- 11478998 TI - Anakinra for arthritis. PMID- 11478999 TI - PCBs on a hook. PMID- 11479001 TI - Dodgy malaria drugs. PMID- 11479002 TI - Opiate withdrawal during development: are NMDA receptors indispensable? AB - Despite decades of research, the mechanisms that underlie opiate tolerance, dependence and withdrawal remain elusive. Evidence accumulated over the past ten years suggests that the NMDA receptor plays a central role in mediating the neuroplasticity induced by chronic opiate administration in adult animals. Yet, during ontogeny, the NMDA receptor complex undergoes qualitative developmental changes, which renders some of the basic assumptions for a role of the NMDA receptor in opiate withdrawal invalid in infants. Recent data indicate that NMDA receptor antagonists are not effective in blocking morphine tolerance, dependence and withdrawal in the neonatal rat. Roles for other glutamate receptor types (e.g. metabotropic glutamate receptors) have also been proposed recently. In this article, the latest evidence that characterizes the dynamic roles of glutamate receptors in these phenomena during ontogeny will be discussed. PMID- 11479003 TI - Therapeutic opportunities from muscarinic receptor research. AB - Muscarinic acetylcholine receptor subtypes have been the subjects of research for at least a quarter of a century. Nonetheless, there are few selective muscarinic receptor ligands presently used as therapeutics. The extensive development of muscarinic M(1) receptor agonists for the treatment of cognitive dysfunction has culminated in a series of unsuccessful drug candidates, which reflects a lack of understanding of the disease and the role played by muscarinic cholinergic transmission. Paradoxically, the most successful antagonist approved for use in urinary incontinence is the nonselective muscarinic receptor antagonist tolterodine. This deficit in subtype-selective ligands could be circumvented by the development of transgenic mice, each lacking functional M(1), M(2), M(3), M(4) or M(5) receptors. In this article, the current status of muscarinic receptor research is critically assessed. PMID- 11479004 TI - Engineering receptors activated solely by synthetic ligands (RASSLs). AB - The functional and molecular diversity of G-protein-coupled receptors presents a significant challenge to understanding the connection between a single receptor signaling pathway and a specific physiological or pathological response. To gain control over the timing and specificity of a G-protein signal, receptors activated solely by synthetic ligands (RASSLs) have been developed. These engineered receptors no longer respond to endogenous peptides, but can still be activated by a specific small-molecule drug. Further control over the location of the signal can be achieved by using RASSLs in conjunction with tissue-specific expression systems in vivo. Existing RASSLs have clarified the role of G(i) signaling in cardiac physiology and are currently being used to study cardiomyopathy, muscle remodeling, sensory transduction and complex neurobehavioral responses. PMID- 11479005 TI - Pharmacogenetic determinants of anti-cancer drug activity and toxicity. AB - Cellular responses to anti-cancer agents result from the interaction between drugs, cellular targets and mechanisms of damage repair. Despite the pharmacological advances in the treatment of cancer, the clinical efficacy of chemotherapy is unpredictable in most patients. However, new information on the genetics of cancer delineates strategies by which the genetic background of tumour cells and patients might be profiled to select anti-cancer agents with improved efficacy and tolerability. This article focuses on the application of pharmacogenetics in the characterization of differences in the pharmacokinetics and pharmacodynamics of anti-cancer agents among individuals to define the likelihood of response and reduce the incidence of adverse effects. PMID- 11479006 TI - DNA microarrays in neuropsychopharmacology. AB - Recent advances in experimental genomics, coupled with the wealth of sequence information available for a variety of organisms, have the potential to transform the way pharmacological research is performed. At present, high-density DNA microarrays allow researchers to quickly and accurately quantify gene-expression changes in a massively parallel manner. Although now well established in other biomedical fields, such as cancer and genetics research, DNA microarrays have only recently begun to make significant inroads into pharmacology. To date, the major focus in this field has been on the general application of DNA microarrays to toxicology and drug discovery and design. This review summarizes the major microarray findings of relevance to neuropsychopharmacology, as a prelude to the design and analysis of future basic and clinical microarray experiments. The ability of DNA microarrays to monitor gene expression simultaneously in a large scale format is helping to usher in a post-genomic age, where simple constructs about the role of nature versus nurture are being replaced by a functional understanding of gene expression in living organisms. PMID- 11479008 TI - Age related signal decrease in functional magnetic resonance imaging during motor stimulation in humans. AB - Right handed healthy volunteers underwent functional magnetic resonance imaging (fMRI) examinations on a 1.5 Tesla MRI-scanner (Gyroscan ACS NT; Philips, Best, NL). Blood oxygen level dependent (BOLD) images were obtained using a three dimensional multi-shot echo planar imaging sequence employing a shifted echo technique (Principles of echo shifting with a train of observations). Finger tapping of the right hand was used as a task for motor stimulation. A total of 86 subjects was included into statistical analysis. Absolute and relative signal differences and cluster sizes of activation for the left motor cortex were obtained. In addition, Z-score, pooled Z-score and cross correlation activation maps were calculated and matched with high resolution anatomic images. A significant decrease with age could be detected for absolute and relative signal intensity differences for the whole group and for the male subgroup. Correlation analysis for the female subgroup also bore negative albeit non-significant correlation coefficients. An age-related decline of BOLD-contrast can be assumed to explain signal decrease. This age-related effect should be considered in clinical fMRI applications. PMID- 11479009 TI - The role of rat dorsomedial prefrontal cortex in working memory for egocentric responses. AB - The present experiment studied the effects of quinolinic acid (90 mM) lesions of the medial precentral and anterior cingulate on working memory for egocentric responses. Rats were trained on a delayed match-to-sample task that involved memory for a 90 degrees right or left turn. The results indicated that lesioned rats had significantly decreased scores during the 10 s delay condition compared to presurgery levels. An increase in the delay to 20 s significantly reduced working memory performance in the lesioned rats compared to that of controls. These findings suggest that the rat medial precentral and/or anterior cingulate area play an important role in working memory for egocentric responses. PMID- 11479010 TI - Impaired retrograde axonal transport of adenovirus-mediated E. coli LacZ gene in the mice carrying mutant SOD1 gene. AB - A replication-defective recombinant adenoviral vector containing E. coli lacZ gene was injected into the gastrocnemius muscles of transgenic mice carrying mutant Cu/Zn superoxide dismutase (SOD1) gene and non-transgenic wild-type mice at 40 weeks of age. After 60 and 90 h of the injection, lacZ staining was observed at the distal ends of the sciatic nerves in both mice groups, with the number and the distances greatly reduced in the transgenic mice. Mean velocities of retrograde transport for lacZ was estimated to be 2.1 and 0.05 mm/24 h in non transgenic and transgenic mice, respectively. These results indicate that the retrograde axonal transport of foreign gene product is impaired in the mice model for familial amyotrophic lateral sclerosis. PMID- 11479011 TI - Opioid, cannabinoid and vanilloid receptor localization on porcine cultured myenteric neurons. AB - Opioids and cannabinoids have profound inhibitory actions on intestinal motility which are mediated in part by their cognate receptors in the enteric nervous system. In the present study, we examined the expression of immunoreactivity for delta- and kappa-opioid receptors, CB(1)-cannabinoid receptors and type 1 vanilloid receptors by immunocytochemistry and confocal laser scanning microscopy on ileal myenteric neurons, isolated from juvenile pigs, that were <70 microm diameter in either axis and maintained for 1-2 weeks in primary culture. Immunoreactivities for delta-opioid and cannabinoid receptors were present in neurons immunoreactive for the cholinergic marker, choline acetyltransferase. Some neurons with delta-opioid receptor-like immunoreactivity were also immunoreactive for kappa-opioid, cannabinoid or vanilloid receptors. These observations indicate that receptors for cannabinoids or vanilloids are co localized in opioid receptor-expressing myenteric neurons which modulate intestinal sensorimotor function. PMID- 11479012 TI - Immunohistochemical study on the distribution of voltage-gated K(+) channels in rat brain following transient focal ischemia. AB - The present study examined the spatial localizations of voltage-gated K(+) (Kv) channels in the rat brain following transient focal ischemia, using immunohistochemistry. Increased expression of Kv1.2 was obvious in the cerebral cortex, dentate gyrus, amygdala, and hypothalamic areas at 3 days following ischemic insults. There was a significant increase in Kv1.2 immunoreactivity in several cortical regions, including cingulate cortex, infralimbic cortex, dorsal peduncular cortex and piriform cortex. On the contrary, Kv1.2 immunoreactivity had not significantly increased in the hippocampal CA1-3 regions although moderate Kv1.2 immunoreactivity was found in the cell bodies and processes of some neurons. Potentially the first demonstration of spatial changes in Kv1.2 channel expression could provide important molecular basis for altered neuronal excitability after ischemic brain injury. PMID- 11479013 TI - Rat tooth pulp cells elicit neurite growth from trigeminal neurones and express mRNAs for neurotrophic factors in vitro. AB - Molecular factors control the developmental ingrowth of axons to the tooth pulp. Here we examine the ability of pulpal cells to induce neurite outgrowth from neonatal rat trigeminal neurones (TGNs) in vitro. We found that TGNs emitted neurites and formed networks of branches in relation to pulpal cells. Neurones co cultured with a mixture of pulpal cells and 3T3 fibroblasts formed networks exclusively in relation to the pulpal cells. Cultivated pulpal cells and pulpal tissue produced mRNAs for all neurotrophins and members of the glial cell line derived neurotrophic factor family. Hence, rat pulpal cells have neuritogenic effects on single TGNs in vitro, that may be associated with secretion of neurotrophic factors. PMID- 11479014 TI - The upregulation of nerve growth factor receptors in reactive astrocytes of rat spinal cord during experimental autoimmune encephalomyelitis. AB - Using immunocytochemistry, we have examined the effect of experimental autoimmune encephalomyelitis (EAE) upon the expression of nerve growth factor (NGF) and its TrkA and p75 receptors in astroglia cells of the spinal cord of Lewis rats. We have found that, in normal spinal cord, astroglia of white matter expressed both NGF receptors while those in gray matter expressed only TrkA and no astroglia expressed NGF. During EAE, strong upregulation of TrkA in the astroglia of gray and white matter was found, particularly in a population of radially oriented astrocytes. An upregulation of p75 was noted in radial astroglia and, to some extent, also in the stellate astrocytes of white matter. In general, the upregulation of NGF receptor immunoreactivities in astroglia correlated with the strong intensification of glial fibrillary acidic protein immunocytochemistry, a prominent feature of EAE. No NGF immunoreactivity appeared in any astroglia cells during EAE. Our results suggest that, during EAE, astroglia of the spinal cord become particularly receptive to NGF, possibly as part of a mechanism enabling astroglial cells to respond to localized release of neurotrophins. Moreover, our data suggest that spinal cord astroglia cells may be a potential target for pharmacological manipulations in EAE. PMID- 11479015 TI - Liposome-mediated gene transfer to fetal human ventral mesencephalic explant cultures. AB - The feasibility of non-viral gene transfer using liposomes is described for human fetal nigral tissue. Ventral mesencephalic explants from 6 to 12 week old fetuses were grown as free-floating roller tube cultures. For the transfection, a vector coding for beta-galactosidase driven by the Rous Sarcoma Virus promoter was used. The developmental stage of the human tissue, time in vitro and the amount of vector DNA used significantly influenced the transfection efficiency. Optimal transfection results were obtained with tissue from a 10 week old fetus, cultured for 4 days and transfected with mixtures containing 4 microg vector DNA. Histological analysis suggested that a specific population of ventral mesencephalic precursor cells were the target for the gene transfer. This finding might have implications for gene delivery and cell replacement strategies in Parkinson's disease. PMID- 11479016 TI - Ethanol induced changes in cyclin-dependent kinase-5 activity and its activators, P35, P67 (Munc-18) in rat brain. AB - The expression of cyclin-dependent kinase-5 (Cdk5) and its regulators, p35 and p67 was investigated in adult rat cerebral cortex and cerebellum, using an experimental paradigm of in vivo chronic ethanol exposure. In parallel, the activity of Cdk5 kinase was measured using a specific substrate histone-H1 peptide. Western blot analysis revealed no appreciable change in the expression of Cdk5 protein levels while, its regulatory proteins, p35 and p67 showed decreased levels following chronic ethanol treatment. However, ethanol treatment resulted in increased Cdk5 activity in both cortex and cerebellum with relatively high activity in cortex. Given the abundant expression and functions of Cdk5 in neural cells, our data implies a regulatory role for Cdk5 in ethanol mediated cell injury and may contribute to impaired CNS development in brain atrophy associated with alcoholic neurodegeneration. PMID- 11479017 TI - Extracellular signal-regulated kinases 1 and 2 phosphorylated neurons in the tele and diencephalon of rat after visceral pain stimulation: an immunocytochemical study. AB - We aimed at verifying whether extracellular signal-regulated kinases (erks) 1 and 2 are activated, i.e. phosphorylated, in forebrain neurons after visceral pain stimulation (VPS). Ether and urethane anaesthetized rats received an intraperitoneal injection of acetic acid or were left untreated (ECT, UCT). After 2 h the animals were perfused. Paraffin embedded brain sections immunoreacted with an antibody selective for the phosphorylated erks. The light microscope analysis revealed only a few labelled neurons in ECT, while in UCT, positive cells were detected. In VPS rats (VPSR) positive cells were mainly distributed in regions, such as the hypothalamic anterior and thalamic paraventricular midline nuclei, amygdala, hippocampal and parahippocampal, insular and perirhinal cortex, involved in nociception and/or visceral activities. Our data suggest an association of erks activation with the emotional component of nociception; moreover, they show that erks activation is not suppressed by anaesthesia. PMID- 11479018 TI - Interpositus nucleus inactivation reduces unconditioned response amplitude after paired but not explicitly unpaired treatment in rabbit eyeblink conditioning. AB - The amplitude of unconditioned responses (URs) in unconditioned stimulus (US) alone presentations were measured in six rabbits during explicitly unpaired and classical conditioning treatments. After both phases of the experiment, the interpositus nucleus (IPN) was reversibly inactivated by a cold probe. URs after unpaired treatment were unaffected by inactivation but after acquisition of a robust level of conditioned responses (CRs), URs in US-alone test trials were reduced in amplitude compared with URs immediately before and after inactivation. The results suggest that the IPN has a role in CR-related reflex modification. PMID- 11479019 TI - Abolished angiogenicity and tumorigenicity of rat glioma by 1 naphthalenemonosulfonate. AB - Suramins and suradistas, an important group of potential anti-cancer agents, inhibit fibroblast growth factor (FGF) mitogenic activity. It has been shown that naphthalenesulfonates, with a common chemical function to the family of suramins and suradistas, mimic their inhibitory activity, abolishing FGF-induced angiogenesis in vivo, and inducing apoptosis of C6 glioma cells in culture. In the present report, we show that intratumoral administration of 1 naphthalenemonosulfonate induces a considerable regression of gliomas in rats, significantly enhances apoptosis, and attenuates tumor angiogenesis. These findings may lead to new approaches for the treatment of glioblastoma, a most common primary malignant brain tumor of very poor prognosis, as well as of other angiogenesis-dependent malignancies. PMID- 11479020 TI - Endogenous adrenergic control of reflexes evoked by mechanical stimulation of the heel in the decerebrated rabbit. AB - In decerebrated rabbits, reflexes were evoked in medial gastrocnemius motoneurones by mechanical stimulation of the heel, using four pinch strengths from 183 (innocuous) to 4577 (noxious) mN. The alpha(2)-adrenoceptor antagonist idazoxan (1-156 microg intrathecal (i.th.) significantly increased responses to pinch strengths of 607 mN and above. Subsequent administration of the alpha(1) adrenoceptor selective antagonist prazosin (200 microg i.th.) decreased reflexes to 4577 mN pinches but had no other significant effects. The opioid antagonist ( )-quadazocine (25 microg i.th.) caused no further changes in reflexes. Spinal section at L1 in the presence of this drug combination enhanced gastrocnemius responses to 183 and 607 mN stimuli, had no effect on reflexes to 1866 mN and significantly decreased responses to 4577 mN pinches. These data confirm that reflexes evoked by 'natural' stimulation of heel mechanoreceptors are subject to powerful tonic descending inhibition mediated by alpha(2)-adrenoceptors. For the highest strength stimulus, the results of alpha(2) blockade involved enabling of descending facilitation as well as reduction of descending inhibition. PMID- 11479021 TI - Early activation of the primary somatosensory cortex without conscious awareness of somatosensory stimuli in tumor patients. AB - The primary sensory cortex has usually been regarded as a necessary step in the information processing stream leading to conscious awareness. Recently, it has been proposed that that higher order associative areas rather than the primary sensory areas are the neural basis of conscious perception. In two patients with tumors near the central region we recorded magnetic somatosensory evoked fields. Magnetic source imaging revealed early (40 ms) neural activation in primary somatosensory cortex and absence of later (>60 ms) neural activation in the primary and associative areas in these patients. None of the patients showed conscious awareness of somatosensory stimuli applied to the corresponding body site although the first component of the evoked field was within normal limits. The time course of the magnetic responses and additional evidence on intensity ratings of somatosensory stimuli suggest that early activity in the primary somatosensory cortex is not sufficient for conscious experience to emerge. PMID- 11479022 TI - Stimulatory effect of harmane and other beta-carbolines on locus coeruleus neurons in anaesthetized rats. AB - Harmane, harmaline and norharmane are beta-carboline related compounds which have been proposed to be endogenous ligands for imidazoline receptors. The effect of these compounds on the activity of locus coeruleus (LC) neurons was studied by extracellular recordings techniques. Intracerebroventricular administration of harmane and harmaline increased the firing rate of LC neurons. Systemic administration of efaroxan, a mixed alpha(2)-adrenoceptor/I(1)-imidazoline antagonist or vagotomy failed to modify the harmane effect. Furthermore, local applications of harmane and harmaline increased the firing rate of LC neurons in a dose-related manner. Finally, intravenous administration of norharmane also increased the activity of LC neurons. Our results demonstrate that beta carbolines stimulate LC neuron activity and indicate that this stimulation occurs directly in the LC by a mechanism independent of I(1)- and I(2)-imidazoline receptors. PMID- 11479023 TI - Enhanced excitatory synaptic transmission in spiny neurons of rat striatum after unilateral dopamine denervation. AB - The synaptic transmission and intrinsic membrane properties of spiny neurons in rat neostriatum were studied after unilateral dopamine depletion using in vivo intracellular recording and staining techniques. Two to four weeks after dopamine denervation, the spontaneous firing rate of spiny neurons increased and the spontaneous membrane potential fluctuation stayed at a more depolarized state for longer periods of time. The amplitude of cortically evoked initial excitatory postsynaptic potentials increased and a late excitatory postsynaptic potential that was occasionally found in control neurons was elicited from 23% of spiny neurons after dopamine denervation. No significant changes in intrinsic membrane properties of spiny neurons were observed after dopamine denervation. These results suggest that dopamine inhibits excitatory synaptic transmission of spiny neurons in naive animals. PMID- 11479024 TI - Local cerebral glucose utilization decreases after heatstroke onset in rats. AB - The quantitative autoradiographic 2-[(14)C]-deoxy-D-glucose methods were used to assess the local cerebral glucose utilization (LCGU) in 46 brain structures in rats suffering from heatstroke. Heatstroke was induced by exposing the animals to an ambient temperature of 42 degrees C. The time at which the local cerebral blood flow (CBF) and mean arterial pressure (MAP) decreased from their peak levels was taken as the onset of heatstroke. Control rats were exposed to a temperature of 24 degrees C. The values of local CBF, MAP and means of total LCGU after heatstroke onset were all significantly lower than those in control rats. However, the values of colonic temperature were greater. Thus, it appears that the decreased cerebral metabolism and perfusion can be due to secondary effect of hypertension after heatstroke onset. PMID- 11479025 TI - Effects of lipofectin-antigen complexes on major histocompatibility complex class I-restricted antigen presentation pathway in murine dendritic cells and on dendritic cell maturation. AB - We previously reported that exogenous antigens complexed with the cationic liposome lipofectin (LF) were efficiently presented via major histocompatibility complex (MHC) class I molecules on pulsed dendritic cells (DCs) in vitro. In the present study, we demonstrated that MHC class I-restricted antigen presentation on DC2.4 cells, a murine immature DC line, treated with LF-antigen complexes was remarkably suppressed through the inhibition of endocytosis, proteasome catalysis, and Golgi transport. We also found that LF did not influence expression of interleukin-12 p40 mRNA, MHC molecules, or co-stimulatory molecules in DC2.4 cells. These findings suggest that an antigen-loading procedure using LF could enhance delivery of exogenous antigens to the classical MHC class I pathway in DCs, but it does not initiate DC maturation. PMID- 11479026 TI - Legume lectin family, the 'natural mutants of the quaternary state', provide insights into the relationship between protein stability and oligomerization. AB - Legume lectins family of proteins, despite having the same 'jelly roll' tertiary structural fold at monomeric level, exhibit considerable variation in their quaternary structure arising out of small changes in their sequence. Nevertheless, their folding behavior and stability correlates very well with their patterns of assembly into dimers and tetramers. A conservation of their fold during evolution, its wide distribution in many protein families together with the availability of structural information on them make them interesting as proteins to explore the effect of inter- versus intra-subunit interactions in the stability of multimeric proteins. Additionally, as 'natural mutants' of quaternary association, proteins of legume lectin family provide interesting paradigms for studies addressing the effect of subunit oligomerization on the stability, folding and function as well as the evolution of multimeric structures. PMID- 11479027 TI - The effect of pH on glucoamylase production, glycosylation and chemostat evolution of Aspergillus niger. AB - The effect of ambient pH on production and glycosylation of glucoamylase (GAM) and on the generation of a morphological mutant produced by Aspergillus niger strain B1 (a transformant containing an additional 20 copies of the homologous GAM glaA gene) was studied. We have shown that a change in the pH from 4 to 5.4 during continuous cultivation of the A. niger B1 strain instigates or accelerates the spontaneous generation of a morphological mutant (LB). This mutant strain produced approx. 50% less extracellular protein and GAM during both chemostat and batch cultivation compared to another strain with parental-type morphology (PS). The intracellular levels of GAM were also lower in the LB strain. In addition, cultivation of the original parent B1 strain in a batch-pulse bioreactor at pH 5.5 resulted in a 9-fold drop in GAM production and a 5-fold drop in extracellular protein compared to that obtained at pH 4. Glycosylation analysis of the glucoamylases purified from shake-flask cultivation showed that both principal forms of GAM secreted by the LB strain possessed enhanced galactosylation (2-fold), compared to those of the PS. Four diagnostic methods (immunostaining, mild methanolysis, mild acid hydrolysis and beta galactofuranosidase digestion) provided evidence that the majority of this galactose was of the furanoic conformation. The GAMs produced during batch-pulse cultivation at pH 5.5 similarly showed an approx. 2-fold increase in galactofuranosylation compared to pH 4. Interestingly, in both cases the increased galactofuranosylation appears primarily restricted to the O-linked glycan component. Ambient pH therefore regulates both GAM production and influences its glycosylation. PMID- 11479028 TI - The preparation of magnetic proteinaceous microspheres using the sonochemical method. AB - Using high-intensity ultrasound, we have developed a method for the synthesis of magnetic microspheres. The microspheres are composed of iron oxide-filled and coated globular bovine serum albumin (BSA). The magnetic microspheres are prepared from BSA and iron pentacarbonyl, or from BSA and iron acetate. Transmission electron microscopy and scanning electron microscopy show spherical particles. The particle size distributions are gaussian, with a mean diameter of a few micrometers. Using chemical analysis, it was found that the total percentage of iron oxide in the microspheres is between 39% and 42%. Mossbauer measurements were also performed. PMID- 11479029 TI - Elongation factor 1beta is an actin-binding protein. AB - A 17 kDa polypeptide found in association with actin in cellular extracts of Dictyostelium discoideum was identified as a proteolytic fragment of eEF1beta. Antibody elicited against the 17 kDa protein reacted with a single 29 kDa polypeptide in Dictyostelium, indicating that the 17 kDa peptide arises from degradation of a larger precursor. The cDNA isolated from a Dictyostelium library using this antibody as a probe encodes Dictyostelium elongation factor 1beta. Amino acid degradation of the 17 kDa protein fragment confirmed the identity of the protein as eEF1beta. Direct interaction of eEF1beta with actin in vitro was further demonstrated in mixtures of actin with the 17 kDa protein fragment of Dictyostelium eEF1beta, recombinant preparations of Dictyostelium eEF1beta expressed in Escherichia coli, and the intact eEF1betagamma complex purified from wheat germ. Localization of eEF1beta in Dictyostelium by immunofluorescence microscopy reveals both diffuse cytoplasmic staining, and some concentration in the cortical and hyaline cytoplasm. The results support the existence of physical and functional interactions of the translation apparatus with the cytoskeleton, and suggest that eEF1beta may function in a dual role both to promote the elongation phase of protein synthesis, and to interact with cytoplasmic actin. PMID- 11479030 TI - Dicynthaurin: an antimicrobial peptide from hemocytes of the solitary tunicate, Halocynthia aurantium. AB - We isolated a novel antimicrobial peptide, dicynthaurin, from hemocytes of a tunicate, Halocynthia aurantium. The native peptide had a mass of approximately 6.2 kDa and was composed of two 30-residue monomers without sequence homology to any previously identified peptides (ILQKAVLDCLKAAGSSLSKAAITAIYNKIT). Most cynthaurin molecules were C-terminally amidated and were linked covalently by a single cystine disulfide bond. When performed in membrane-mimetic environments, circular dichroism studies of dicynthaurin revealed largely alpha-helical conformations. Dicynthaurin's broad-spectrum activity encompassed Gram-positive (Micrococcus luteus, Staphylococcus aureus, Listeria monocytogenes) and Gram negative bacteria (Escherichia coli, Pseudomonas aeruginosa), but not Candida albicans, a fungus. Although dicynthaurin was purified from a marine invertebrate, its antimicrobial activity was optimal at NaCl concentrations below 100 mM. This suggests that the antimicrobial actions of this molecule may take place intracellularly (e.g., within a phagosome) rather than extracellularly. PMID- 11479031 TI - Cell age-related monovalent cations content and density changes in stored human erythrocytes. AB - Conversion of erythrocyte membrane protein 4.1b to 4.1a occurs through a non enzymatic deamidation reaction in most mammalian erythrocytes, with an in vivo half-life of approximately 41 days, making the 4.1a/4.1b ratio a useful index of red cell age [Inaba and Maede, Biochim. Biophys. Acta 944 (1988) 256-264]. Normal human erythrocytes distribute into subpopulations of increasing cell density and cell age when centrifuged in polyarabinogalactan density gradients. We have observed that, when erythrocytes were stored at 4 degrees C under standard blood bank conditions, the deamidation was virtually undetectable, as cells maintained the 4.1a/4.1b ratio they displayed at the onset of storage. By measuring the 4.1a/4.1b values in subpopulations of cells of different density at various time points during storage, a modification of the normal 'cell age/cell density' relationship was observed, as erythrocytes were affected by changes in cell volume in an age-dependent manner. This may stem from a different impact of storage on the imbalance of monovalent cations, Na(+) and K(+), in young and old erythrocytes, related to their different complement of cation transporters. PMID- 11479032 TI - In situ observation of the generation of isothiocyanates from sinigrin in horseradish and wasabi. AB - Sulfur K-edge X-ray absorption spectroscopy has been used to determine the chemical identity of the sulfur-containing species in horseradish (Armoracia lapthifolia) and wasabi (Wasabia japonica) in situ, before and after cell disruption. The major sulfur-containing species in the intact root is sinigrin (1 thio-beta-D-glucopyranose 1-N-(sulfoxy)-3-buteneimidate) and related congeners. Disrupting the cells by applying local pressure allowed the conversion of the sulfur moieties in sinigrin to isothiocyanates and sulfate in approximately equimolar amounts. In contrast to previous suggestions, no detectable thiocyanates were formed, but an unusual thio intermediate may have been identified for the first time. PMID- 11479033 TI - Oxygen free radical generating mechanisms in the colon: do the semiquinones of vitamin K play a role in the aetiology of colon cancer? AB - It is proposed that bile acids (deoxycholic acid), the K vitamins, iron(II) complexes and oxygen interact to induce an oncogenic effect in the colon by the generation of free radicals. In the relatively low oxidising/reducing conditions of the colonic lumen the K vitamins exist in the reduced form; however, if absorbed into the mucosa they have the capacity to be chemically oxidised and to enter into a redox cycle yielding oxygen radicals. The semiquinone radical of K(1) (phylloquinone) has been stabilised in bile acid mixed micelles and investigated by electron paramagnetic resonance spectroscopy and quantum chemical calculations. The estimated half-life of the radical was about 30 min which confirms a remarkably high stability in aqueous micellar solution. A model is presented in which the reduced K vitamins may initiate superoxide radical, O2(-*) generation leading to Fe(II) mediated Fenton reactions in the stem colon cells. PMID- 11479034 TI - Enzyme activities involved in tryptophan metabolism along the kynurenine pathway in rabbits. AB - The following enzyme activities of the tryptophan-nicotinic acid pathway were studied in male New Zealand rabbits: liver tryptophan 2,3-dioxygenase, intestine indole 2,3-dioxygenase, liver and kidney kynurenine 3-monooxygenase, kynureninase, kynurenine-oxoglutarate transaminase, 3-hydroxyanthranilate 3,4 dioxygenase, and aminocarboxymuconate-semialdehyde decarboxylase. Intestine superoxide dismutase and serum tryptophan were also determined. Liver tryptophan 2,3-dioxygenase exists only as holoenzyme, but intestine indole 2,3-dioxygenase is very active and can be considered the key enzyme which determines how much tryptophan enters the kynurenine pathway also under physiological conditions. The elevated activity of indole 2,3-dioxygenase in the rabbit intestine could be related to the low activity of superoxide dismutase found in intestine. Kynurenine 3-monooxygenase appeared more active than kynurenine-oxoglutarate transaminase and kynureninase, suggesting that perhaps a major portion of kynurenine available from tryptophan may be metabolized to give 3 hydroxyanthranilic acid, the precursor of nicotinic acid. In fact, 3 hydroxyanthranilate 3,4-dioxygenase is much more active than the other previous enzymes of the kynurenine pathway. In the rabbit liver 3-hydroxyanthranilate 3,4 dioxygenase and aminocarboxymuconate-semialdehyde decarboxylase show similar activities, but in the kidney 3-hydroxyanthranilate 3,4-dioxygenase activity is almost double. These data suggest that in rabbit tryptophan is mainly metabolized along the kynurenine pathway. Therefore, the rabbit can also be a suitable model for studying tryptophan metabolism in pathological conditions. PMID- 11479035 TI - Malignancies and the skin: looking forward and looking back. PMID- 11479036 TI - Cutaneous malignancies of keratinocytic origin. PMID- 11479037 TI - Cutaneous malignant melanoma. PMID- 11479038 TI - Cutaneous and subcutaneous sarcomas. PMID- 11479039 TI - Kaposi's sarcoma. PMID- 11479040 TI - Cancer-associated genodermatoses and familial cancer syndromes with cutaneous manifestations. PMID- 11479041 TI - Cutaneous manifestations of internal malignancies. PMID- 11479042 TI - Medical management of cutaneous malignancies. PMID- 11479043 TI - Surgical management of cutaneous malignancies. PMID- 11479044 TI - Cryosurgery in the management of cutaneous malignancies. PMID- 11479045 TI - Laser and photodynamic therapy in the management of cutaneous malignancies. PMID- 11479046 TI - Mohs surgery in the management of cutaneous malignancies. PMID- 11479047 TI - Radiation therapy in the management of cutaneous malignancies. PMID- 11479048 TI - Total skin electron beam therapy in the management of cutaneous malignancies. PMID- 11479049 TI - The psychological comorbidity in acne. PMID- 11479050 TI - Research directions in genetic counselling: a review of the literature. AB - There is a growing body of literature considering genetic counselling services in a variety of clinical settings. This literature encompasses both predictive and diagnostic testing, from the viewpoints of service providers and recipients. It also embraces a wide range of conceptions of the nature and goals of genetic counselling. However, research in this area has been criticised for a focus on outcome rather than process, and it has been suggested that this focus limits its practical use. The purpose of this review is twofold: (1) to describe the varying concepts of counselling which appear to be utilised in published work and (2) to discuss the possible applications of this work to practice. PMID- 11479051 TI - Chronic disease patient education: lessons from meta-analyses. AB - Twelve meta-analyses were identified concerning education for people with chronic diseases where behaviour modification is a part of the treatment regime. By combining the results of these meta-analyses a second stage descriptive meta analysis was conducted. The aim of the exercise was to explore the effects of patient education and implications for educational treatment. The results provided evidence of the gaps that exist in current research practice. These gaps include the need to utilise rigorous research designs to explore the quantitative effects of patient education, and the need to qualitatively explore the processes by which these effects have transpired. Where randomised controlled trials had been conducted the effects of patient education were usually small and were only known for 6 months of follow-up. In addition, the educational interventions tested were generally poorly described, and failed to adhere to theoretical models. The results of this review have highlighted the need for practitioners to use theoretically based teaching strategies which include behaviour change tactics that affect feelings and attitudes. In alignment with these conclusions, the review has provided guidelines for future research practice. PMID- 11479052 TI - The RE-AIM framework for evaluating interventions: what can it tell us about approaches to chronic illness management? AB - BACKGROUND: The RE-AIM framework is used as a method of systematically considering the strengths and weaknesses of chronic illness management interventions in order to guide program planning. METHOD: The RE-AIM dimensions of Reach, Efficacy, Adoption, Implementation, and Maintenance are used to rate one-on-one counseling interventions, group sessions, interactive computer mediated interventions, telephone calls, mail interventions, and health system policies. RESULTS: The RE-AIM ratings suggest that, although often efficacious for those participating, traditional face-to-face intervention modalities will have limited impact if they cannot be delivered consistently to large segments of the target population. Interventions using new information technologies may have greater reach, adoption, implementation, and maintenance, and thereby greater public health impact. Policy changes received high ratings across a variety of RE AIM dimensions. CONCLUSIONS: Program planners should make decisions regarding implementing and funding health services based on multiple dimensions, rather than only considering efficacy in randomized clinical trials. Doing so may improve the resulting public health impact. Directions for future chronic illness management research related to RE-AIM, and implications for decision making, are described. PMID- 11479053 TI - Field test of a group education program for type 2 diabetes: measures and predictors of success on individual and group levels. AB - We performed field testing of a previously described group education program for type 2 diabetes. HbA(1c) levels at start, 6 and 12 months were collected and demographic factors examined to identify predictors of long-term glycemic control on individual and group levels. "Glycemic success" comprised of (1) achieving target values of HbA(1c) < or =6.5% and/or (2) decreasing HbA(1c) progressively, depending on initial values. Groups in the field test and previous pilot-study (N=105) decreased their mean HbA(1c) significantly after 6 months, implying that diabetes mass education led by pharmacists could be possible in the future. Target HbA(1c) < or =6.5% was seen in 51% at start and 63% after 12 months (P=0.023). Initial HbA(1c) and BMI were the most important success predictors; age, sex, duration, and civil status showed no effects. Overweight individuals relapsed after initially decreasing their HbA(1c), emphasizing the need for long term support in weight management. Experienced loneliness affected outcomes, indicating interaction between diet self-care and social relations. PMID- 11479054 TI - Pediatric injury prevention counseling: an observational study of process and content. AB - OBJECTIVES: To describe routine injury prevention counseling; to observe how three visit components - printed prompts, parent remarks, and parent behaviors - affect such counseling; to describe the process and content of discussions about car seats as an example of routine injury prevention. METHODS: A total of 128 well-child visits of children under 7 months of age to a university pediatric clinic were videotaped (76% of eligible visits). RESULTS: Three injury topics were mentioned, on an average, per visit. Parents or caregivers rarely introduced injury topics (5%). Physicians frequently introduced those topics listed on age specific prompting sheets (73%). Car seat counseling typically began with a physician's question (82%). Most asked simply about ownership or use (93%). Few addressed difficult issues, such as consistency of use (11%). CONCLUSIONS: Physicians bring up the injury topics that are prompted. However, most discussion is superficial. Printed prompts that address counseling process as well as content might be beneficial. PMID- 11479055 TI - Parents: the best experts in child health care? Viewpoints from parents and staff concerning child health services. AB - The aim was to describe what parents and staff think about child health care, to identify agreements and disagreements. A qualitative study was made with semi structured interviews based on a phenomenographic approach. Sixty parents, 14 nurses and six doctors from southern Sweden were interviewed. Parents and staff emphasized two tasks as being of particular importance: support and check-ups. There was a conflict between parents' need for security versus integrity. Individual nurses experienced a conflict between what they wanted to do and what they felt that they had to do. The parents viewed parental education as a chance to exchange experiences with other parents and receive support from other adults, while the staff mainly saw it as an opportunity to inform parents and strengthen them in their parental role. The study gives grounds for reflection about how the work of child health care can be changed in the future. PMID- 11479056 TI - Patients' understandings of heart attack: implications for prevention of recurrence. AB - Patients' willingness to undertake secondary preventive strategies following heart attack are likely to be affected by their understandings of their condition. This qualitative study explored patients' understandings of heart attack in order to contribute to the design of effective secondary prevention services. In-depth interviews were conducted with 25 patients with myocardial infarction (MI). These data indicate that information received from health professionals encourages patients to view heart attack as an acute event rather than a symptom of a chronic condition and that this understanding provides patients with low motivation for long-term lifestyle change. Patients may benefit from understanding a heart attack as an acute symptom of an underlying disease process which long-term medication and behavioural change can help to check. In order to achieve this, health professionals need to examine patients' understandings of their heart attack and recovery and to provide information about lifestyle which engages with these understandings. PMID- 11479057 TI - Evaluation of a psycho-educational nutritional program in diabetic patients. AB - In this study, we have retrospectively (2 years) assessed the efficacy of the nutritional education (NE) program provided to 83 diabetic patients during an in hospital training week. Two years after NE, type 1 diabetic patients have decreased significantly their total caloric intake, protein (P<0.05) and fat consumption. Type 2a diabetic patients disclosed a significant increase in CHO consumption while type 2b diabetic patients decreased their fat consumption, SFA consumption and total caloric intake. The percentage of patients following the European Association Study for Diabetes (EASD) nutritional recommendations regarding fat intake has increased significantly in all three diabetic groups and in type 2a diabetic patients regarding CHO consumption. In conclusion, a psycho educational nutritional education program of 1 week allows a significant improvement of the alimentary behavior 2 years thereafter with 45% of the diabetic patients following the current EASD dietary recommendations. PMID- 11479058 TI - Advance directives in dementia care: from instructions to instruments. AB - Advance directives are often regarded as instructions to the doctor about future care. This view is problematic, in that it obliterates that decisions about treatment and care always take place in a concrete situation, and require interpretation and communication. From a hermeneutic perspective, advance directives can be regarded as instruments which do not replace interpretation and communication, but sustain joint decision-making about treatment and care, including the patient and the family in a process of meaning-making. PMID- 11479059 TI - Development and evaluation of a multi-family psychoeducational program for cystic fibrosis. AB - The development, implementation and evaluation of a psychoeducational program for families with a child affected by cystic fibrosis (CF) is described. Aim of the program was to strengthen the families' coping with CF-related problems and to improve adherence with chest physiotherapy. Sixteen families from an outpatient CF-clinic participated. Parents and children were educated both individually as well as together in multi-family groups. Teaching, practicing and group discussions were balanced in each session. In a pre-post-design the following variables were evaluated: parental coping, parental health beliefs, children's coping, adherence, and knowledge about CF. The parental coping patterns and health beliefs remained unchanged in the study group. Children developed more search for social support, whereas their competence and optimism decreased and withdrawal increased slightly as perceived by the parents. There is a subgroup with poor family functioning and adherence at pretest which improves after the intervention.Family-centered psychoeducational intervention may be a promising supportive strategy for children with CF, especially if it is dedicated to families with poor adaptation to the disease. PMID- 11479061 TI - Preventing severe mental illnesses--new prospects and ethical challenges. AB - We review some exciting recent developments in studies of psychoses in general and schizophrenia in particular that offer hope such illnesses can be identified and treated early. Early treatment promises better outcomes for both the affected individuals and their families, and perhaps even the prospect of true prevention. But testing such interventions in vulnerable patients can raise ethical difficulties that if not adequately addressed may harm patients and hamper the development of these interventions. We believe there are appropriate ways of dealing with the ethical problems in the early intervention field, though we also suggest that additional work on both scientific and ethical fronts will need to be done to make these ways real. As a start toward that end, we have summarized a number of the debates in the field and from the following articles, and have distilled five 'points to consider' that interested parties will need to take into account. PMID- 11479062 TI - Ethics and early intervention in psychosis: keeping up the pace and staying in step. AB - The intense clinical and research interest in early psychosis in recent years has highlighted a range of ethical issues which need to be considered carefully. Our perspective is based on 16 years of clinical and research experience with young people at this phase of illness as well as the research contributions of many others. We discuss the ethical dilemmas in relation to the three key foci, which make up the early psychosis paradigm. These are the pre-psychotic or prodromal phase, the period of untreated psychosis and the first psychotic episode and the critical period of recovery, which follows this. Most attention is devoted to the pre-psychotic period, however ethical considerations related to research in the other two clinical foci are briefly covered as well. Our contention is that the ethical issues are essentially identical to those arising in early intervention research in mainstream medicine. This has been concealed by inconsistency and emotion, which has great potential to confuse, politicize and derail rational debate. The legacy of the isolation of psychiatry from medicine and consequent prejudice and stigma in the professional as well as the public mind seems to be fueling a tendency in some societies to view psychiatric research as qualitatively different from other medical research. Sound clinical research data should be allowed to illuminate the options for potential consumers across all phases of illness. The alternative is research paralysis, which would force clinical practice to expand blindly without an evidence base. PMID- 11479063 TI - Treatment of the schizophrenia prodrome: is it presently ethical? AB - Although the devastating consequences of schizophrenia have long been known, interest in preventive intervention has only recently emerged. The shift in focus toward early treatment has been encouraged by findings suggesting that the longer psychosis remains untreated, the poorer the prognosis, and by the recent introduction of novel antipsychotic medications with a more benign side effect profile than conventional neuroleptics. In this paper, we argue that interest in prevention has outpaced the necessary scientific and ethical underpinnings for clinical trials involving the schizophrenia prodrome. Specifically, we maintain that the prodromal phase of schizophrenia is, at present, essentially a retrospective construct and that, as a result, the defining signs and symptoms currently in use must be validated in naturalistic, longitudinal studies. In particular, it is essential to establish solid base rates for schizophrenia in prodromal individuals before early treatment can be effectively evaluated. Additional ethical/scientific issues discussed include: (1) the need for an exit strategy (i.e. the determination of when to discontinue treatment in an individual who does not develop schizophrenia), (2) the advisability of pharmacological interventions that specifically target neurocognitive deficits, and (3) the possibility that antidepressant medications may be as effective or more effective, with fewer side effects, than antipsychotic medication for prodromal individuals. PMID- 11479064 TI - Early detection strategies for untreated first-episode psychosis. AB - Some studies in first-episode schizophrenia correlate shorter duration of untreated psychosis (DUP) with better prognosis, suggesting that timing of treatment may be important. A three-site prospective clinical trial in Norway and Denmark is underway to investigate the effect of the timing of treatment in first episode psychosis. One health care sector (Rogaland, Norway) is experimental and has developed an early detection (ED) system to reduce DUP. Two other sectors (Ulleval, Norway, and Roskilde, Denmark) are comparison sectors and rely on existing detection and referral systems for first-episode cases. The study ultimately will compare early detected with usual detected patients. This paper describes the study's major independent intervention variable, i.e. a comprehensive education and detection system to change DUP in first onset psychosis. System variables and first results from the four-year inclusion period (1997-2000) are described. It includes targeted information towards the general public, health professionals and schools, and ED teams to recruit appropriate patients into treatment as soon as possible. This plus easy access to psychiatric services via ED teams systematically changed referral patterns of first-episode schizophrenia. DUP was reduced by 1.5 years (mean) from before the time the ED system was instituted (to 0.5 years). The ED strategies appear to be effective and to influence directly the community's help-seeking behaviour. PMID- 11479065 TI - Psychosis treatment prior to psychosis onset: ethical issues. AB - Clinical trials have begun of antipsychotic treatments in persons who are prodromally symptomatic and at high risk for schizophrenia but who have not yet become psychotic. The ethical issues connected with intervening prior to making a diagnosis of psychosis are detailed. Compelling but tentative evidence suggests that early treatment may improve course and prognosis, and this has initiated a paradigm shift in thinking about the risks and benefits of early intervention. The nature of this evidence, its implications, its shortcomings, and its effect upon the ethics of treating schizophrenia are elaborated and discussed. It is concluded that clinical psychiatry is currently in a state of "equipoise" or genuine uncertainty about the comparative merits of early treatment, a state which endorses early intervention research, including intervention in the prodromal phase. PMID- 11479066 TI - Early intervention and a five year follow up in young adults with a short duration of untreated psychosis: ethical implications. AB - In a Dutch treatment intervention study of patients (n=76) with first psychotic episodes of schizophrenia the hypothesis tested was whether early differential treatment after an acute psychotic break improved outcome as compared with other studies. Patients had a relatively short duration of untreated psychosis. No significant effect between two treatment conditions on relapse rate was found. The 15-month intervention program kept the psychotic relapse rate as low as 15%; lower than comparable studies. Thus, the initial results were in support of the hypothesis. After completion of the 15 months study, patients were referred to other agencies and followed for five years. Results of the follow up study showed that the low relapse rate could not be maintained. Of the remaining 71 patients of the initial sample, 52% had one or more psychotic relapses, 25% developed chronic positive symptoms and 23% did not have another psychotic episode. In addition, the level of social functioning turned out to be low: the majority of patients were dependent upon their parents, few held down a skilled or paid job and also their quality of life seemed low, results indicate that early intervention may improve short term but not long term outcome in schizophrenia. Our results also suggest that referral to other mental health agencies after intervention is not sufficient. Continuity of outpatient care, including continuity of a professional relationship, continuity of support for the family, and the continuity in management of illness, medication and stress may be a key issue in the first five years after the onset of psychosis in schizophrenia. Early recognition and intervention may not nearly be as important for outcome as continuity in care and caregivers. At present, however, it remains questionable whether early intervention programs in first-episode patients with a short duration of untreated psychosis can offer the prospect of altering the course of schizophrenia without a sustained comprehensive treatment program. PMID- 11479067 TI - Dutch law and ethics concerning the experimental treatment of early psychosis. AB - Moral and social attitudes towards medical treatment and research rapidly changed during the last four decades of the Twentieth Century. In the Netherlands these changes are reflected in three laws adopted in the 1990s. The Law on Medical Treatment Agreement (Wet Geneeskundige BehandelOvereenkomst, WGBO, 1992) reflects the respect for autonomy and the protection of patient's rights. The Law on Special Admission to Psychiatric Hospitals (Bijzondere Opneming Psychiatrische Ziekenhuizen, BOPZ, 1994) reflects the idea that psychiatric subjects should be entitled to execute their civil rights as long as no danger exists to themselves or others, while the Law on Medical Research (Wet Medisch Onderzoek, WMO 1999), reflects the present moral opinions on bio-medical research. From these laws objections can be derived against the development of treatment and experiments on early psychosis. It will be argued here that these laws can also be taken to stimulate this kind of research. The difference between the two options is related to preconceived moral opinions towards psychosis and schizophrenia. PMID- 11479068 TI - Rationale for the study of early intervention. AB - Consistent but relatively weak evidence exists that treating patients with schizophrenia early in the course of their illness can decrease long-term morbidity. Relatedly, it is possible that treating individuals even earlier might produce better results. The findings presented set the stage for early and even earlier formal intervention studies, where the potential benefits are thought to outweigh the potential risks. PMID- 11479069 TI - Ethical issues in early-intervention clinical trials involving minors at risk for schizophrenia. AB - Early-intervention clinical trials involving persons at risk for schizophrenia raise a variety of ethical issues. These issues range from some of the most general questions of ethical theory and research ethics to some very specific pragmatic questions about optimal procedures for protecting subjects. Giving special attention to minor subjects (and adult subjects lacking decision-making capacity), this paper aims to provide an overview of some leading ethical issues and concerns as well as a rights-based framework and several concrete suggestions for addressing these issues and concerns. PMID- 11479070 TI - International clinical trials with applicability to mentally impaired individuals: the conundrum of third world sites. AB - Research on the effectiveness of treatment programs in early stages of severe mental illness raise significant ethical questions when conducted in industrialized countries that have adequate protections for the human subjects of biomedical and behavioral research. Before embarking on similar trials in developing countries, researchers face formidable barriers that must first be addressed and overcome. The challenges to external sponsors of research in conducting ethically responsible clinical trials in developing countries include at least the following: the existence of alternative belief systems regarding the causes and treatment of mental illness; difficulty in implementing the results of successful research in resource-poor countries; providing for adequate prior ethical review in the host country of the proposed study; and problems in ensuring that voluntary, informed consent is properly obtained from research subjects or their legally authorized representatives. Global justice demands that we not adopt one ethical standard for rich countries and another for resource poor countries. PMID- 11479071 TI - Family and twin strategies as a head start in defining prodromes and endophenotypes for hypothetical early-interventions in schizophrenia. AB - In an effort to share the experiences of 'genotype-hunters'-who have approached the difficult task of forecasting future schizophrenia in the young offspring or other relatives of index cases, in new samples guided by the prior probabilities of 15% in offspring or 50% in identical co-twins-with 'early-interventionists' who focus on purported prodromal symptoms in children who would be treated pharmacologically to prevent the development of schizophrenia-we provide a focused review that emphasizes the hazards of false positives in both approaches. Despite the advantages prospective high-risk strategies have had from clinical and laboratory findings that implicate some prodromal signs and endophenotypes, e.g. attention, memory, and information processing evaluations, the yields are not sufficient for practical applications involving antipsychotic drugs for undiagnosed children. Even more caution than usual is required, given the suggestions that the developing neocortex is vulnerable to dopaminergic exposure. PMID- 11479072 TI - Preventing schizophrenia and Alzheimer disease: comparative ethics. AB - Schizophrenia and Alzheimer disease are both diseases of the brain that involve genetic susceptibility factors and for which the prevention or delay of symptom onset are important research goals. This paper provides some comparisons between current preventive efforts in schizophrenia and Alzheimer disease, focusing on certain ethical features of these endeavors such as potential discrimination, misdiagnosis, and stigma. PMID- 11479073 TI - An overview of women's health issues in the United States and United Kingdom. PMID- 11479074 TI - National perspective: United States. PMID- 11479075 TI - National perspective: United Kingdom. PMID- 11479076 TI - Women's primary health care and the search for quality. PMID- 11479077 TI - Women's primary health care and the search for quality. PMID- 11479082 TI - Patient-centered decision making: empowering women to make informed choices- experience in the United States. PMID- 11479083 TI - Patient-centered decision making: empowering women to make informed choices. PMID- 11479088 TI - Improving access and quality for ethnic minority women. PMID- 11479089 TI - Improving access and quality for ethnic minority women. PMID- 11479094 TI - Domestic violence as a women's health issue. PMID- 11479095 TI - Domestic violence as a women's health issue. PMID- 11479100 TI - When teens talk "chastity," have them make the commitment. PMID- 11479101 TI - The medical evaluation in cases of child sexual abuse. PMID- 11479102 TI - Plasma carnitine levels of pregnant adolescents in labor. AB - STUDY OBJECTIVE: To determine the concentration of plasma carnitine (total, free, and acylcarnitine) during the delivery of uncomplicated pregnancies of adolescent women. To investigate the relationship between maternal and neonatal levels of carnitine and to compare these carnitine levels between pregnant and nonpregnant adolescents. DESIGN: Samples of maternal and umbilical blood were taken at the time of delivery and examined for the determination of the carnitine-total, free, and acylcarnitine-concentration by the use of an enzymatic-radioisotope method. Twenty-two cases of uncomplicated adolescent pregnancies with a normal labor and without perinatal complications were examined. The plasma level of carnitine was also examined in 17 healthy nonpregnant adolescent women, which constituted the control group. RESULTS: The concentrations of plasma carnitine in adolescent pregnancies at the time of delivery were calculated at 19.6 +/- 2.15 microMol/L (total), 12.62 +/- 1.31 microMol/L (free), and 6.98 +/- 1.55 microMol/L (acylcarnitine). The corresponding mean values in umbilical plasma were 30.31 +/- 2.06 microMol/L, 22.39 +/- 1.64 microMol/L, and 7.92 +/-.96 mucroMol/L. There is a statistically significant difference between the mean values in maternal and umbilical plasma (P <.0001 for total and free carnitine and P <.012 for acylcarnitine). The correlations between adolescent pregnant women and their infants as regards total, free, and acylcarnitine were 0.137, 0.018, and 0.33, respectively. Neither of these parameters was statistically significant. The corresponding mean values of carnitine in nonpregnant adolescent women were statistically significantly higher than in adolescent pregnant women (total carnitine: 41.61 +/- 3.09 microMol/L, free: 31.39 +/- 2.81 microMol/L, acylcarnitine: 10.22 +/- 1.88 microMol/L, P <.0001). CONCLUSIONS: The concentration of plasma carnitine at the end of adolescent pregnancy is low compared to the levels of umbilical carnitine at birth and that found in nonpregnant adolescent women. It may not have an obvious impact on the utilization of fatty acids in an uncomplicated full-term pregnancy; however, it suggests the potential risk for neonatal fatty-acid oxidation in a preterm or complicated pregnancy. PMID- 11479103 TI - Mood changes in adolescents using depot-medroxyprogesterone acetate for contraception: a prospective study. AB - STUDY OBJECTIVE: Depot-medroxyprogesterone acetate (DMPA) is thought to cause changes in mood among patients using it for contraception. The purpose of this study was to evaluate changes in negative and positive affect among adolescent females using DMPA as a contraceptive agent. DESIGN, SETTING, PARTICIPANTS: This prospective study was set in an urban hospital adolescent clinic. Thirty-nine adolescents choosing DMPA as a contraceptive agent and 24 adolescents not using any hormonal contraception were enrolled as subjects and controls, respectively. Two standardized questionnaires, the Beck Depression Inventory (BDI) and the Multiple Affect Adjective Checklist-Revised (MAACL-R), were administered at baseline to all participants and readministered at 3, 6, and 12 months. MAIN OUTCOME MEASURES: Changes in negative affect from baseline to 3, 6, and 12 months were evaluated by the BDI and by "dysphoria" subscale scores of the MAACL-R. Paired t-tests were used to measure these changes in subjects and controls separately. RESULTS: The mean change in BDI scores from baseline to one year for those who completed one year was -4.8 for subjects (P =.02) and +.3 (P =.84) for controls. The mean change in the dysphoria subscale scores was -5.7 (P =.21) for the subjects and -.1 (P =.98) for the controls while the change in the positive affect scores over a period of one year were -2.1 (P =.46) and +.1 (P =.98) for subjects and controls, respectively. CONCLUSIONS: Adolescents using DMPA do not show depressive symptoms when using DMPA as a contraceptive agent over a period of 12 months as measured by the BDI and show no significant changes in negative or positive affect as measured by the MAACL-R. PMID- 11479104 TI - 47,XXX in an adolescent with premature ovarian failure and autoimmune disease. AB - BACKGROUND: Premature ovarian failure (POF) may be idiopathic or may be associated with genetic or autoimmune disorders. The 47,XXX karyotype has been associated with POF and other genitourinary anomalies. CASE: A 17-year-old woman with a history of immune thrombocytopenic purpura was referred to the adolescent medicine clinic for evaluation of oligomenorrhea with secondary amenorrhea. Evaluation revealed hypergonadotrophic premature ovarian failure, a positive antinuclear antibody, and the 47,XXX karyotype. She has since developed a positive anti-cardiolipin antibody but does not meet diagnostic criteria for systemic lupus erythematosis. CONCLUSION: The presence of known autoimmune disease in a woman with POF should not dissuade the clinician from evaluating for a potential genetic cause. PMID- 11479105 TI - Comparison of microscopic examination and human papillomavirus DNA subtyping in vulvar lesions of premenarchal girls. AB - STUDY OBJECTIVE: The purpose of this study is to compare the microscopic examination and human papillomavirus (HPV) DNA subtyping of vulvar specimens from premenarchal girls clinically diagnosed with condyloma to determine whether DNA subtyping aids in the diagnostic process. DESIGN: A retrospective chart review was performed on all premenarchal girls who underwent surgical treatment of clinically diagnosed condyloma between 1993 and 1999 at the University of Michigan Medical Center by the Pediatric and Adolescent Gynecology Service. Tissue was sent for pathologic evaluation and in 10 patients the specimens also underwent DNA subtyping. One patient had prior DNA subtyping. All the other lesions were surgically ablated. The microscopic slides were reviewed by a single pathologist blinded to the study. SETTING: The study was performed in a tertiary care university hospital. PARTICIPANTS: The study group included 11 premenarchal girls with an average age of 2.3 yr. MAIN OUTCOME MEASURES: The charts were reviewed for previous HPV treatment, maternal history of HPV, history of sexual abuse, microscopic diagnosis, and HPV DNA subtyping. RESULTS: Four patients had prior surgical treatment and two patients had undergone prior medical treatment. The microscopic diagnosis was condyloma in 8 patients, chronic dermatitis in 2 patients, and 1 patient had VIN 2-3. All 11 specimens tested positive for HPV DNA, 10 specimens contained at least one of the low-risk subtypes (6, 11, 42, 43, 44), and 1 tested positive for low-risk as well as intermediate/high-risk HPV subtypes (16, 18, 31, 33, 35, 45, 51, 52, 56). CONCLUSIONS: Although all the patients with a clinical diagnosis of condyloma tested positive for HPV DNA, only 9 of 11 were definitely diagnosed with HPV-related pathology by microscopic examination. Therefore, in premenarchal patients with verrucous lesions in the anogenital area, microscopic evaluation alone may be inadequate as a confirmatory test when a positive clinical diagnosis has been made, and HPV DNA subtyping should be considered to avoid confusion with the diagnosis. PMID- 11479106 TI - Adolescent girl's coping with an STD. Not enough problem solving and too much self-blame. AB - STUDY OBJECTIVE: Approximately three million teenagers are infected with an STD each year. The ways in which an adolescent girl copes with an STD may have implications for future risk and for psychological adjustment. The purpose of the current study was to compare whether coping with an STD was similar to coping with other stressors. SETTING: Urban, hospital-based adolescent medicine clinic. DESIGN AND PARTICIPANTS: Sixty-seven girls with a mean age of 15.9 (sexual debut was 13.8) yr completed the KIDCOPE in response to both an STD acquisition and an interpersonal stressor within the previous 6 months. RESULTS: Problem solving was used less often, and self-blame was used more often, in response to an STD acquisition. Frequency of use of self-blame was not correlated with perceived helpfulness. CONCLUSIONS: These findings suggest that clinicians need to help adolescent girls manage STD acquisition from the perspective of problem solving rather than self-blame. PMID- 11479107 TI - Sexual abuse knowledge base among residents in family practice, obstetrics/gynecology, and pediatrics. AB - STUDY OBJECTIVE: To investigate resident physician knowledge about sexual abuse prevalence and understanding about potential perpetrators. DESIGN: Questionnaires were mailed to program directors in family practice, obstetrics and gynecology, and pediatric residency programs. PARTICIPANTS: The questionnaires were distributed to senior residents in their final months prior to graduation. INTERVENTIONS: Residents were asked to fill out the questionnaire anonymously and return it to our institution in the prepaid envelope provided. MAIN OUTCOME MEASURES: Demographic characteristics and knowledge of sexual abuse prevalence and perpetrator characteristics were assessed. Chi-square contingency table analysis was used to compare responses of the three specialties. RESULTS: The overwhelming majority (98.8%) of residents correctly identified a family member as the individual most likely to sexually abuse a child. Approximately half of the residents knew the correct prevalence of sexual abuse among females and among males. There was a weak understanding of the potential youthfulness of juvenile offenders. CONCLUSION: We believe that resident understanding of sexual abuse prevalence and about the youthfulness of juvenile offenders can be improved in all three specialties. PMID- 11479108 TI - Hirsutism investigations-what is appropriate? PMID- 11479109 TI - Management quandary. Testicular feminization. PMID- 11479110 TI - A role for the nurse practitioner. PMID- 11479112 TI - Combinatorial chemistry. Web alert. AB - A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Chemical Biology. PMID- 11479114 TI - Techniques for analysis and purification in high-throughput chemistry. AB - The success of combinatorial chemistry, and the increased emphasis on single well characterised compounds of high purity, has had a significant impact on analytical and purification technologies. The requirement for ever-increasing throughput has led to the automation and parallelisation of these techniques. Advances have also been made in developing faster methods to augment throughput further. PMID- 11479115 TI - Combinatorial approaches to affinity chromatography. AB - A new armoury of protein purification tools is required to support rapid advances in high-throughput genomics and proteomics, which are predicted to lead to the discovery, isolation, characterisation and manufacture of a number of new biopharmaceutical proteins. Computer-aided molecular design, combinatorial (bio)chemistry and high-throughput screening techniques are now being exploited to identify highly selective ligands for use in the purification of these proteins by affinity chromatography. PMID- 11479116 TI - Solutions for library encoding to create collections of discrete compounds. AB - The ability to design, produce, analyze, and manage high quality combinatorial libraries depends on the encoding strategy applied. Several recent advances in encoding technology have extended the range of library design parameters. Enhancements of established techniques have made them more robust and versatile, while additional new and creative encoding concepts have been introduced. With better options now available, combinatorial chemists can more easily select an encoding technique to match resources, library design, and compound management criteria. PMID- 11479117 TI - Quantitative techniques for the comparison of solid supports. AB - The host of solid supports available to the synthetic chemist adds yet another level of complexity to solid-phase synthesis. Although the selection of the optimal solid support for a specific synthetic transformation is still empirically driven, significant progress has been made in the development of quantitative techniques to compare solid supports, providing new insight into the microenvironment created by the interaction of the solid support with solvent. PMID- 11479118 TI - Combinatorial electrochemistry. AB - Recent advances in combinatorial electrochemistry include new methods for parallel construction of small organic molecules, synthesis of conducting materials, electroactive solid supports and heterogeneous catalysts. In addition, new instrumentation for high-throughput electrosynthesis and analysis continues to be developed at a steady pace. Future development is likely to involve miniaturization of a variety of electrosynthetic operations. PMID- 11479119 TI - Lead compounds discovered from libraries. AB - Lead compounds with the potential to progress to viable drug candidates have been identified from libraries using several strategies. These include rapid screening of large diverse collections, thematic libraries, project-directed libraries, and three-dimensional molecular models of corporate databases. There have been numerous success stories, including the identification of several clinical candidates. PMID- 11479120 TI - Applications of NMR in drug discovery. AB - NMR, already some 50 years old, has long been an invaluable analytical method in industry for verification of chemical synthesis and compound characterisation. The range of molecular information accessible through NMR, however, offers a far larger horizon of applications. Of these, ligand screening by NMR has emerged as a very promising new method in drug discovery. Its unmatched screening sensitivity, combined with the abundance of available information on the structure and nature of molecular binding, justifies the growing interest in this dynamically expanding NMR application. PMID- 11479121 TI - Natural-product-like chiral derivatives by solid-phase synthesis. AB - In this genomics and proteomics age, highly functionalized natural products or natural-product-like compounds are likely to play important roles in understanding the functions of emerging biological targets because they serve as small-molecule chemical probes in modulating a target's specific actions (i.e. activation or deactivation). Development of stereoselective reaction-derived methods on solid phase provides a means of obtaining functionalized chiral core structures that may be used for high-throughout syntheses. PMID- 11479122 TI - Combinatorial alanine-scanning. AB - Combinatorial libraries of alanine-substituted proteins can be used to rapidly identify residues important for protein function, stability and shape. Each alanine substitution examines the contribution of an individual amino acid sidechain to the functionality of the protein. The recently described method of shotgun scanning uses phage-displayed libraries of alanine-substituted proteins for high-throughput analysis. PMID- 11479123 TI - Molecular addresses in blood vessels as targets for therapy. AB - We have isolated several organ- and tumor-homing peptides by using in vivo phage display. This technology involves the screening of peptide libraries in a living animal. The peptides that result from such a selection home to specific organs or tissues because they recognize molecular 'addresses', receptors that are differentially expressed in vascular beds. Targeted delivery of chemotherapeutics, pro-apoptotic peptides and cytokines to tumors using these peptides improved therapeutic efficacy in animal models. Translation of this technology into clinical applications will form the basis for targeting therapeutic and imaging agents in the context of cancer and other diseases. PMID- 11479124 TI - Random-peptide libraries and antigen-fragment libraries for epitope mapping and the development of vaccines and diagnostics. AB - Random peptide libraries and antigen-fragment libraries (also known as gene fragment libraries) have been used to identify epitopes on protein antigens. These technologies promise to make significant contributions to diagnostic and vaccine development. Researchers in a number of labs have shown that phage selected from libraries with protective antibodies, raised against whole antigen, can be used as immunogens to stimulate antibody responses that bind native antigen and provide protection in vivo. Others have used the sera of patients with idiopathic diseases to screen libraries, and by this approach have identified candidate antigens involved in immune disease. These may prove useful for diagnosis and, possibly, in determining disease etiology. PMID- 11479125 TI - Combinatorial syntheses of sugar derivatives. AB - There has been an exponential growth in interest of the functional roles of carbohydrates and cell surface glycoconjugates in the past 10 years. The importance of glycoconjugates as mediators of biosignals has stimulated investigation into simple and versatile methods for their synthesis. The synthesis of carbohydrates and glycoconjugates by combinatorial chemistry has gained considerable interest. PMID- 11479127 TI - Using green fluorescent protein to study intracellular signalling. AB - Subcellular compartmentalisation of signalling molecules is an important phenomenon not only in defining how a signalling pathway is activated but also in influencing the desired physiological output of that pathway (e.g. cell growth or differentiation, regulation of metabolism, cytoskeletal changes etc.). Biochemical analyses of protein and lipid compartmentalisation by, for example, subcellular fractionation presents many technical difficulties. However, this aspect of cell signalling research has seen a major revolution thanks to the cloning and availability of a variety of mutant green fluorescent protein derivatives with distinct molecular properties. Mutants with increased brightness, altered excitation and emission maxima, altered stability and differential sensitivity to pH, are now in widespread use for following the trafficking and function of proteins in living cells and for monitoring the intracellular environment. In this review we focus on some of the recent developments in the use of green fluorescent proteins for studying intracellular signalling pathways often with special reference to the actions of insulin. We also discuss the future utility of these proteins to analyse protein--protein interactions in signalling pathways using fluorescence resonance energy transfer. PMID- 11479128 TI - Up to date with human thyroglobulin. AB - The coding region of the human thyroglobulin (TG) mRNA has been resequenced, and comparison with the TG sequence originally published in 1987 showed many variations. All of the variations were validated in 20--40 other alleles, and this resulted in the revision of 41 nucleotide positions. This review presents the revised wild-type human TG sequence, including all known exon/exon boundaries and additional data on the TG mRNA population, concerning alternative splicing and variability of the polyadenylation cleavage site. The amino acid sequence derived shows one additional, 12 changed, and 10 polymorphic residues. Protein characteristics, such as acceptor and donor tyrosine residues, N-glycosylation sites, cysteine-rich repeats, the proposed receptor domain, and antigenic epitopes, are included, and their relationship to the revised sequence is discussed. Furthermore, all reported TG mutations causing dyshormonogenesis in humans and animals are designated in the nucleotide and amino acid sequences. This up-to-date profile of the human TG molecule presents the features of importance for its complex role in thyroid hormonogenesis, and is the basis for future studies on the structure--function relationship. PMID- 11479129 TI - Dysregulation of the adipoinsular axis -- a mechanism for the pathogenesis of hyperleptinemia and adipogenic diabetes induced by fetal programming. AB - Obesity and its related disorders are the most prevalent health problems in the Western world. Using the paradigm of fetal programming we developed a rodent model which displays the phenotype of obesity and metabolic disorders commonly observed in human populations. We apply maternal undernutrition throughout gestation, generating a nutrient-deprived intrauterine environment to induce fetal programming. Maternal undernutrition results in fetal growth retardation and in significantly decreased body weight at birth. Programmed offspring develop hyperphagia, obesity, hypertension, hyperleptinemia and hyperinsulinism during adult life and postnatal hypercaloric nutrition amplifies the metabolic abnormalities induced by fetal programming. The adipoinsular axis has been proposed as a primary candidate for linking the status of body fat mass to the function of the pancreatic beta-cells. We therefore investigated the relationship between circulating plasma concentrations of leptin and insulin and immunoreactivity in the endocrine pancreas for leptin and leptin receptor (OB-R) in genetically normal rats that were programmed to become obese during adult life. Virgin Wistar rats were time mated and randomly assigned to receive food either available ad libitum (AD group) or at 30% of the ad libitum available intake (UN group). Offspring from UN mothers were significantly smaller at birth than AD offspring (AD 6.13+/-0.04 g, UN 4.02+/-0.03 g, P<0.001). At weaning, offspring were assigned to one of two diets (a standard control diet or a hypercaloric diet consisting of 30% fat) for the remainder of the study. At the time of death (125 days of age), UN offspring had elevated (P<0.005) fasting plasma insulin (AD control 1.417+/-0.15 ng/ml, UN control 2.493+/-0.33 ng/ml, AD hypercaloric 1.70+/-0.17 ng/ml, UN hypercaloric 2.608+/-0.41 ng/ml) and leptin (AD control 8.8+/-1.6 ng/ml, UN control 14.32+/-1.9 ng/ml, AD hypercaloric 15.11+/-1.8 ng/ml, UN hypercaloric 30.18+/-5.3 ng/ml) concentrations, which were further increased (P<0.05) by postnatal hypercaloric nutrition. The elevated plasma insulin and leptin concentrations were paralleled by increased immunolabeling for leptin in the peripheral cells of the pancreatic islets. Dual immunofluorescence histochemistry for somatostatin and leptin revealed that leptin was co-localized in the pancreatic delta-cells. OB-R immunoreactivity was evenly distributed throughout the pancreatic islets and was not changed by programming nor hypercaloric nutrition. Our data suggest that reduced substrate supply during fetal development can trigger permanent dysregulation of the adipoinsular feedback system leading to hyperleptinemia, hyperinsulinism and compensatory leptin production by pancreatic delta-cells in a further attempt to reduce insulin hypersecretion in the progression to adipogenic diabetes. PMID- 11479130 TI - The role of the IGF axis in IGFBP-1 and IGF-I induced renal enlargement in Snell dwarf mice. AB - Insulin-like growth factor (IGF) binding protein-1 (IGFBP-1) is generally believed to inhibit IGF action in the circulation. In contrast, IGFBP-1 has been reported to interact with cell surfaces and enhance IGF-I action locally in some tissues. Renal IGFBP-1 levels are found elevated in various conditions characterized by renal growth (e.g. diabetes mellitus, hypokalemia). To test whether IGFBP-1 is a renotropic factor, IGFBP-1 was administered alone or in combination with IGF-I to Snell dwarf mice, an in vivo model without compensatory feedback effects on growth hormone (GH) secretion. In three control groups of Snell dwarf mice, placebo, GH or IGF-I was administered. Compared with placebo, kidney weight increased in all treated groups, however, with different effects on kidney morphology. Administration of IGF-I, alone or in combination with IGFBP-1, tended to increase glomerular volume, while no changes were seen in the other groups. Administration of IGFBP-1 or IGFBP-1+IGF-I both caused dilatation of the thin limbs of Henle's loop, while GH or IGF-I administration had no visible effect. Furthermore, IGF-I administration resulted in an increased mean number of nuclei per cortical area and renal weight, whereas GH, IGF-I+IGFBP-1 or IGFBP-1 caused a decreased renal nuclei number. In situ hybridization and immunohistochemistry showed specific changes of the renal IGF system expression patterns in the different groups. Particularly, IGFBP-1 administration resulted in extensive changes in the mRNA expression of the renal IGF system, whereas the other administration regimen resulted in less prominent modifications. In contrast, administration of IGFBP-1 and IGFBP-1+IGF-I resulted in identical changes in the protein expression of the renal IGF system. Our results indicate that IGFBP-1, alone or in combination with IGF-I, demonstrated effects on the renal tubular system that differ from the effects of IGF-I. PMID- 11479131 TI - Hormonal regulation of tight junction closure in the mouse mammary epithelium during the transition from pregnancy to lactation. AB - Closure of the tight junctions of the mammary epithelium has been shown to accompany the onset of copious milk secretion or lactogenesis, stage 2, in both goats and humans. Here we use injection of [(14)C]sucrose and FITC-albumin (fluorescein isothiocyanate-albumin) into the mammary duct to follow the course of tight junction closure during lactogenesis in mice. To examine the hormonal changes responsible, we ovariectomized day 16 or 17 pregnant mice and found that closure followed ovariectomy with a mean delay of 13.6+/-1.5 (s.e.m. ) h. That progesterone withdrawal is the trigger for closure was shown by the finding that injection of progesterone within 4 h of ovariectomy delayed closure and that closure occurred after injection of the progesterone antagonist RU 486 in intact late pregnant mice. Endocrine ablation studies showed that low to moderate concentrations of corticosterone and either placental lactogen or prolactin are necessary for tight junction closure triggered by progesterone withdrawal. Thus the hormonal requirements for tight junction closure are similar to those shown by other investigators to promote lactogenesis, stage 2. Further, the tight temporal control of tight junction permeability suggests that ovariectomy of the late pregnant mouse may be a good model for molecular studies of the lactogenic switch. PMID- 11479132 TI - Developmental changes in galanin in lumbosacral sympathetic ganglionic neurons innervating the avian uterine oviduct and galanin induction by sex steroids. AB - We recently found lumbosacral sympathetic ganglionic galanin neurons innervating the quail uterine oviduct. Galaninergic innervation of the uterine muscle may be essential for avian oviposition, as galanin evoked oviposition through a mechanism of induction of vigorous uterine contraction. The questions arising from these findings are: what changes occur in galanin expression in the sympathetic ganglionic galanin neuron during development, and what is the hormonal factor(s) that induces galanin expression in this neuron? Therefore, the present study examined the developmental changes in galanin of the quail sympathetic ganglionic neuron and uterus, and the effect of administration of ovarian sex steroids on galanin induction. Immature birds reared under long-day photoperiods from 4 weeks of age demonstrated progressive increases in galanin levels both per unit ganglionic protein (concentration) and per ganglia (content) concurrent with ganglionic development during weeks 4--13. The uterine galanin content and uterine weight also increased progressively during the same period, but the galanin concentration in the uterus at 4 weeks was high due to the much smaller tissue mass. Immunocytochemical analysis with anti-galanin serum showed that immunoreactive ganglionic cells were few and small at 4 weeks and increased progressively thereafter. Administration of oestradiol-17 beta to immature birds at 3 weeks of age for 1 week increased both the galanin concentration and content in the ganglia without ganglionic growth. A marked increase in galanin immunoreactive ganglionic cells was detected following oestradiol treatment. In contrast, progesterone increased ganglionic galanin levels, but the effects were low. Expression of the mRNAs encoding oestrogen receptor-alpha and -beta (ER alpha and ER beta) in the ganglionic tissue was verified by RT-PCR/Southern blot analysis. Immunocytochemical staining with anti-ER serum further revealed an intense immunoreaction restricted to the nucleus of ganglionic neurons. These results suggest that ovarian sex steroids, in particular oestradiol-17 beta, contribute as hormonal factors to galanin induction, which takes place in the lumbosacral sympathetic ganglionic neurons innervating avian uterine oviduct during development. Oestradiol may act directly on this ganglionic neuron through intra-nuclear receptor-mediated mechanisms to induce galanin. PMID- 11479133 TI - Production and localisation of angiotensin II in the bovine early corpus luteum: a possible interaction with luteal angiogenic factors and prostaglandin F2 alpha. AB - The newly formed corpus luteum (CL) rapidly develops after ovulation and has the features of active vascularisation and mitosis of steroidogenic cells. These stage-specific mechanisms also may contribute to gain the function of prostaglandin F2 alpha (PGF2 alpha)-resistant CL at this stage. Recent studies suggest that the vasoactive peptide angiotensin II (Ang II) regulates luteal function. Thus, this study aimed to investigate (i) the expression of angiotensin converting enzyme (ACE) mRNA by RT-PCR and the ACE protein expression by immunohistochemistry, (ii) the effects of angiogenic growth factors, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), on the secretion of Ang II, PGF2 alpha, progesterone and oxytocin (OT), and (iii) the effects of luteal vasoactive peptides (Ang II and endothelin-1 (ET-1)) or OT on the secretion of PGF2 alpha, progesterone and OT from bovine early CL (days 3- 4 of the oestrous cycle), and evaluate a possible interaction of these substances with PGF2 alpha. The expression of mRNA for ACE was found in theca interna of mature follicle, early CL and endothelial cells from developing CL as well as pituitary and kidney, but granulosa cells of mature follicle were negative. The immunohistochemical analysis revealed that blood capillaries (endothelial cells) were stained for ACE, but luteal cells were negative in early CL. To examine the effects of substances on the secretory function of the CL, an in vitro microdialysis system was used as a model. The infusion of bFGF and VEGF stimulated Ang II and PGF2 alpha secretion as well as progesterone, but not OT secretion in early CL. The infusion of Ang II after PGF2 alpha infusion continued the stimulatory effect on progesterone and OT release within early CL until 3 h thereafter. However, the infusion of ET-1 alone had no effect on progesterone or OT release. The infusion of luteal peptides such as Ang II and OT stimulated PGF2 alpha secretion, whereas the infusion of ET-1 did not. In conclusion, the overall results of this study indicate that a functional angiotensin system exists on the endothelial cells of early CL, and that angiogenic factors bFGF and VEGF upregulate luteal Ang II and PGF2 alpha secretion, which fundamentally supports the mechanism of progesterone secretion in bovine early CL. This idea supports the concept that the local regulatory mechanism involved in active angiogenesis ensures the progesterone secretion in the developing CL in vivo. PMID- 11479134 TI - Effects of tri-iodothyronine on alternative splicing events in the coding region of cytochrome P450 aromatase in immature rat Sertoli cells. AB - Transient postnatal hypothyroidism in male rats induces a prolonged proliferation of immature Sertoli cells. This change in Sertoli cell replication at young ages is coincident with enhanced and prolonged aromatase activity that leads to a marked increase in the conversion of androgens into estrogens. Both events are drastically inhibited by tri-iodothyronine (T(3)) replacement either in vivo or in vitro. This study, after the immunolocalization of aromatase in cultured rat Sertoli cells, examined the effects elicited by T(3) on this enzyme, by simultaneously investigating three functional levels of aromatase: mRNA expression, protein content, and enzymatic activity. The immunolocalization of cytochrome P450 aromatase (P450 arom) was shown in the cytoplasm of cultured Sertoli cells from 15- and 21-day-old rats. Western blot analysis revealed an enhancement of aromatase protein content upon stimulation with N(6),2'-O dibutyryladenosine-3':5'-cyclic monophosphate ((Bu)(2)cAMP) that was clearly down regulated by T(3). The presence of a functional P450 arom protein in purified Sertoli cells was confirmed by the measurement of [(3)H]H(2)O released after incubation with [1 beta-(3)H]androst-4-ene-3,17-dione. With 100 nM T3, a decrease in both P450 arom mRNA levels and aromatase activity was observed. The aromatase enzymatic activity was strongly stimulated by (Bu)(2)cAMP and markedly down regulated by T(3). In contrast, the strong increase in aromatase mRNA upon (Bu)(2)cAMP stimulation was apparently unaffected by T(3) administration. This paper shows how the identification of an altered transcript induced by T(3) coding for putative truncated and inactive aromatase protein might explain such a decrease in aromatase activity in T(3)-treated cells. On the basis of these results, it is concluded that at least two mechanisms could be involved in the down-regulatory effect of T(3) on aromatase activity in prepuberal Sertoli cells. The first mechanism is linked to a possible direct modulatory role for T(3) in the regulation of the aromatase promoter, whilst the second one is represented by the induction of altered transcripts coding for truncated and inactive aromatase proteins. PMID- 11479135 TI - Platelet-derived growth factor (PDGF) and PDGF receptors in rat corpus cavernosum: changes in expression after transient in vivo hypoxia. AB - Platelet-derived growth factor (PDGF) overactivity has been implicated in atherosclerosis and several fibrotic conditions including lung and kidney fibrosis, liver cirrhosis and myelofibrosis. Low oxygen tension (hypoxia) is a known stimulus for transcriptional induction of PDGF ligand and receptor genes in different tissues. We studied the expression and localization of PDGF-A, PDGF-B, and PDGF receptor (PDGFR)-alpha and -beta subunits in adult rat isolated corpus cavernosum (CC) under generalized transient hypoxia (pO(2) 10%) in comparison with normoxic conditions. Semi-quantitative RT-PCR analysis of mRNA extracted from rat penis showed higher amounts of PDGF-A, PDGF-B and PDGFR-beta mRNA transcripts in hypoxic versus normoxic animals. The immunohistochemical analysis showed that the localization of PDGF subunits and PDGFR-beta was confined to the cytoplasm of the perivascular smooth muscle cells, endothelium and trabecular fibroblasts. Our findings indicate that transient low oxygen tension induces PDGF overexpression in rat CC, which in the long term may lead to an increase of connective tissue production. We suggest that a local impairment of the PDGF/PDGFR system may contribute to CC fibrosis, which is an established cause of erectile dysfunction in man. PMID- 11479136 TI - LH/chorionic gonadotropin signaling pathway involves protein tyrosine phosphatase activity downstream of protein kinase A activation: evidence of an obligatory step in steroid production by Leydig cells. AB - Our recent reports indicate that protein tyrosine phosphorylation is an obligatory component of the mechanism of action of ACTH in its stimulatory action of corticosteroid production in adrenal zona fasciculata (ZF). The role of protein tyrosine phosphatase (PTP) activity in the regulation of steroidogenesis by LH/chorionic gonadotropin (CG) was tested using cell-permeable PTP inhibitors. Thus, PTP inhibition blocks LH- and 8-bromo-cAMP-stimulated testosterone production by Leydig cells without affecting 22(R)OH-cholesterol-supported steroidogenesis, similar results to those obtained in the adrenal ZF/ACTH system, leading us to propose that PTP action is an obligatory and common step in the cascade triggered by both hormones. Then, we continued the study testing whether LH modulates PTP activity in MA-10 cells, a Leydig cell line. In this regard, we observed by an in-gel PTP assay two PTPs of 110 and 50 kDa that are activated by hormone and 8-bromo-cAMP activation of the cells. Moreover, there is a transient increase by the second messenger in total PTP activity that correlates with the higher activity displayed by the 110 and 50 kDa proteins in the in-gel assay. In accordance with these results, analysis of tyrosine phosphorylated proteins showed the LH-induced dephosphorylation of proteins of 120, 68 and 50 kDa. The results of this study indicate that PTPs play an important role in the regulation of Leydig cell functions and that there exists a cross talk between serine/threonine phosphorylation and tyrosine dephosphorylation mediated by hormone-activated cAMP-dependent protein kinase and PTPs. These results are the first evidence of PTP having a role in LH/CG-stimulated steroidogenesis. PMID- 11479137 TI - Molecular mechanisms of leptin action in adult rat testis: potential targets for leptin-induced inhibition of steroidogenesis and pattern of leptin receptor messenger ribonucleic acid expression. AB - Leptin, the product of the ob gene, is a pivotal signal in the regulation of neuroendocrine function and fertility. Although much of the action of leptin in the control of the reproductive axis is exerted at the hypothalamic level, some direct effects of leptin on male and female gonads have also been reported. Indeed, recent evidence demonstrated that leptin is able to inhibit testosterone secretion at the testicular level. However, the molecular mechanisms behind this effect remain unclear. The focus of this study was twofold: (1) to identify potential targets for leptin-induced inhibition of steroidogenesis, and (2) to characterize in detail the pattern of expression and cellular distribution of leptin receptor (Ob-R) mRNA in adult rat testis. In pursuit of the first goal, slices of testicular tissue from adult rats were incubated with increasing concentrations of recombinant leptin (10(-9)--10(-7 )M) in the presence of human chorionic gonadotropin (hCG; 10 IU/ml). In this setting, testosterone secretion in vitro was monitored, and expression levels of mRNAs encoding steroidogenic factor 1 (SF-1), steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage enzyme (P450 scc) and 17 beta-hydroxysteroid dehydrogenase type III (17 beta-HSD) were assessed by Northern hybridization. In pursuit of the second goal, the pattern of cellular expression of the Ob-R gene in adult rat testis was evaluated by in situ hybridization using a riboprobe complementary to all Ob-R isoforms. In addition, testicular expression levels of the different Ob-R isoforms, previously identified in the hypothalamus, were analyzed by means of semi-quantitative RT-PCR. In keeping with our previous data, recombinant leptin significantly inhibited hCG-stimulated testosterone secretion. In this context, leptin, in a dose-dependent manner, was able to co-ordinately decrease the hCG-stimulated expression levels of SF-1, StAR and P450 scc mRNAs, but it did not affect those of 17 beta-HSD type III. In situ hybridization analysis showed a scattered pattern of cellular expression of the Ob-R gene within the adult rat testis, including Leydig and Sertoli cells. In addition, assessment of the pattern of expression of Ob-R subtypes revealed that the long Ob-Rb isoform was abundantly expressed in adult rat testis. However, variable levels of expression of Ob-Ra, Ob-Re, and Ob-Rf mRNAs were also detected, whereas those of the Ob-Rc variant were nearly negligible. In conclusion, our results indicate that decreased expression of mRNAs encoding several up-stream elements in the steroidogenic pathway may contribute, at least partially, to leptin induced inhibition of testicular steroidogenesis. In addition, our data on the pattern of testicular expression of Ob-R isoforms and cellular distribution of Ob R mRNA may help to further elucidate the molecular mechanisms of leptin action in rat testis. PMID- 11479138 TI - Retinoic acid and vitamin D(3) powerfully inhibit in vitro leptin secretion by human adipose tissue. AB - Leptin, the product of the ob gene, is secreted into the circulation by white adipose tissue; its major role being to participate in the regulation of energy homeostasis. Plasma leptin levels are mainly determined by the relative adiposity of the subject; however, the great dispersion of values for any given body mass index and the noteworthy gender-based differences indicate that other factors are operating. Steroid hormones actively participate in the regulation of leptin secretion; however, non-steroid nuclear hormones have either not been studied or have provided contradictory results. In order to understand the role of hormones of the non-steroid superfamily such as 3,5,3'-tri-iodothyronine (T(3)), vitamin D(3) and retinoic acid (RA) in the control of leptin secretion, in the present work doses of 10(-9), 10(-8) and 10(-7) M of these compounds have been studied on in vitro leptin secretion. The organ culture was performed with omental adipose tissue samples from healthy donors (n=28). T(3) was devoid of effect at any dose studied, while an inhibition of leptin secretion was observed with 9-cis-RA (slight) and all-trans-RA (potent). Interestingly, vitamin D(3) exerted a powerfully inhibitory role at the doses studied, and its action was synergistic with all-trans-RA. In conclusion, in vitro leptin secretion by human adipose tissue is negatively controlled by either RA or vitamin D(3). The clinical significance of leptin regulation by this superfamily of nuclear receptors remains to be ascertained. PMID- 11479139 TI - Nitric oxide as a second messenger in parathyroid hormone-related protein signaling. AB - Parathyroid hormone (PTH)-related protein (PTHrP) is produced in smooth muscles and endothelial cells and is believed to participate in the local regulation of vascular tone. No direct evidence for the activation of endothelium-derived nitric oxide (NO) signaling pathway by PTHrP has been found despite attempts to identify it. Based on direct in situ measurements, it is reported here for the first time that the human PTH/PTHrP receptor analogs, hPTH(1--34) and hPTHrP(1- 34), stimulate NO release from a single endothelial cell. A highly sensitive porphyrinic microsensor with a response time of 0.1 ms and a detection limit of 1 nmol/l was used for the measurement of NO. Both hPTH(1--34) and hPTHrP(1--34) stimulated NO release at nanomolar concentrations. The peak concentration of 0.1 micromol/l hPTH(1--34)- and 0.1 micromol/l hPTHrP(1--34)-stimulated NO release was 175+/-9 and 248+/-13 nmol/l respectively. This represents about 30%--40% of maximum NO concentration recorded in the presence of (0.1 micromol/l) calcium ionophore. Two competitive PTH/PTHrP receptor antagonists, 10 micromol/l [Leu(11),d -Trp(12)]-hPTHrP(7--34)amide and 10 micromol/l [Nle(8,18),Tyr(34)] bPTH(3--34)amide, were equipotent in antagonizing hPTH(1--34)-stimulated NO release; [Leu(11),d -Trp(12)]-hPTHrP(7--34)amide was more potent than [Nle(8,18),Tyr(34)]-bPTH(3--34)amide in inhibiting hPTHrP(1--34)-stimulated NO release. The PKC inhibitor, H-7 (50 micromol/l), did not change hPTH(1--34)- and hPTHrP(1--34)-stimulated NO release, whereas the combined effect of 10 micromol/l of the cAMP antagonist, Rp-cAMPS, and 50 micromol/l of the calmodulin inhibitor, W-7, was additive. The present studies show that both hPTH(1--34) and hPTHrP(1- 34) activate NO production in endothelial cells. The activation of NO release is through PTH/PTHrP receptors and is mediated via the calcium/calmodulin pathway. PMID- 11479140 TI - Effect of estrogen on calcium and sodium transport by the nephron luminal membranes. AB - Estrogens are widely used for contraception and osteoporosis prevention. The aim of the present study was to investigate the effect of 17 beta-estradiol on calcium (Ca(2+)) transport by the nephron luminal membranes, independently of any other Ca(2+)-regulating hormones. Proximal and distal tubules of rabbit kidneys were incubated with 17 beta-estradiol or the carrier for various periods of time, and the luminal membranes of these tubules were purified and vesiculated. Ca(2+) uptake by membrane vesicles was measured using the Millipore filtration technique. Incubation of proximal tubules with the hormone did not influence Ca(2+) uptake by the luminal membranes. In contrast, incubation of distal tubules with 10(-8) M 17 beta-estradiol for 30 min decreased the initial uptake of 0.5 mM Ca(2+) from 0.34+/-0.04 (s.e.m. ) to 0.17+/-0.04 pmol/microg per 5 s (P<0.05). In the presence of 100 mM Na(+), 0.5 mM Ca(2+) uptake was strongly diminished and the effect of 17 beta-estradiol disappeared (0.17+/-0.01 and 0.21+/-0.07 pmol/microg per 5 s in vesicles from the control and treated tubules). Direct incubation of the membranes with 17 beta-estradiol, however, failed to show any influence of the hormone on Ca(2+) transport. The action of 17 beta-estradiol was dose-dependent, with a half-maximal effect at approximately 10(-9) M. Ca(2+) uptake by the distal tubule membranes presents dual kinetics. 17 beta-Estradiol decreased the V(max) value of the high-affinity component from 0.42+/-0.02 to 0.31+/-0.03 pmol/microg per 10 s (P<0.02). In contrast with the effect of the hormone on Ca(2+) transport, estradiol increased Na(+) uptake by both the proximal and distal tubule luminal membranes. In conclusion, incubation of proximal and distal tubules with estrogen decreases Ca(2+) reabsorption by the high-affinity Ca(2+) channels of the distal luminal membranes, and enhances Na(+) transport by the membranes from proximal and distal nephrons. PMID- 11479141 TI - Changing RANKL/OPG mRNA expression in differentiating murine primary osteoblasts. AB - Osteoblast-osteoclast coordination is critical in the maintenance of skeletal integrity. The modulation of osteoclastogenesis by immature cells of the osteoblastic lineage is mediated through receptor activator of NF kappa B (RANK), its ligand RANKL, and osteoprotegerin (OPG), a natural decoy receptor for RANKL. Here, the expression of OPG and RANKL in primary mouse osteoblastic cultures was investigated to determine whether the osteoclastogenic stimulus depended on the stage of osteoblastic differentiation and the presence of the calciotrophic hormone 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)). OPG mRNA expression was increased in osteoblastic cultures after the onset of mineralisation relative to less mature cultures, but did not alter in response to 1,25-(OH)(2)D(3) treatment. In contrast, basal RANK L mRNA expression did not change during differentiation but was significantly enhanced by 1,25-(OH)(2)D(3) treatment at all times. The stimulatory effects of 1,25-(OH)(2)D(3) on RANKL were lessened in more mature cultures, however. The RANKL/OPG ratio, an index of osteoclastogenic stimulus, was therefore increased by 1,25-(OH)(2)D(3) treatment at all stages of osteoblastic differentiation, but to a lesser degree in cultures after the onset of mineralisation. Thus the 1,25-(OH)(2)D(3)-driven increase in osteoclastogenic potential of immature osteoblasts appears to be mediated by increased RANKL mRNA expression, with mature osteoblasts having relatively decreased osteoclastogenic activity due to increased OPG mRNA expression. These findings suggest a possible mechanism for the recently proposed negative regulatory role of mature osteoblasts on osteoclastogenesis and indicate that the relative proportions of immature and mature osteoblasts in the local microenvironment may control the degree of resorption at each specific bone site. PMID- 11479142 TI - Regulation of glucogenesis by thyroid hormones in fetal sheep during late gestation. AB - The effects of thyroid hormone deficiency in utero on the fetal glucogenic capacity were investigated by measuring glucose production and hepatic levels of glycogen and gluconeogenic enzymes in normal sheep fetuses in the fed and fasted states during late gestation and in those made thyroid hormone deficient by fetal thyroidectomy (TX). In the fed state, fetal TX had no effect on glucose uptake, utilisation or production by the fetus. It also had no apparent effect on the glycogen content or activities of the key gluconeogenic enzymes in the fetal liver. In addition, fetal plasma concentrations of insulin, cortisol, adrenaline or noradrenaline were unaffected by fetal TX in the fed state. In contrast, the rates of fetal O(2) consumption and CO(2) production per kilogram fetal bodyweight were significantly lower in TX than in intact fetuses in the fed state (P<0.05). TX prevented fetal glucose production in response to maternal fasting for 48 h. It also abolished the normal decreases in the fetal glucose carbon oxidation fraction, the rate of CO(2) production from glucose carbon and in the fraction of the umbilical O(2) uptake used for glucose carbon oxidation that occur during fasting in intact fetuses. At the end of the fast, plasma noradrenaline concentrations and hepatic levels of glycogen, glucose 6 phosphatase, fructose diphosphatase and alanine aminotransferase were significantly lower in TX than in intact fetuses. These observations show that thyroid hormones are essential for glucogenesis in the sheep fetus during late gestation and suggest that these hormones act both on the hepatic glucogenic pathways and on the mechanisms activating glucogenesis in utero. PMID- 11479143 TI - Interaction of interleukin-6 on human granulosa cell steroid secretion. AB - Numerous studies have shown that a variety of cytokines are involved in the regulation of ovarian function. Interleukin-6 (IL-6), has been found in follicular fluid. In this report we show by immunocytochemical methods the localization of the extracellular domain of the IL-6-receptor and its associated signal transducer glycoprotein gp 130 on the surface of granulosa cells (GCs). The possibility that IL-6 may also influence the basal and FSH-stimulated production of estradiol (E2) and progesterone (Prog) by GCs in vitro was also investigated. GCs were obtained from infertile patients undergoing in vitro fertilization and embryo transfer treatment and cultured for 72 h or were given increasing concentrations of human recombinant IL-6 (8--128 pg/ml) in the absence or presence of FSH (96 U/ml). For the time-course studies, FSH-stimulated GCs were treated in the absence or presence of IL-6 (128 pg/ml) and supernatants were assayed at 24 h intervals (24-96 h) for E2 and Prog productions. The results show that increasing amounts of IL-6 significantly inhibit E2 production in the absence or presence of FSH vs untreated controls (P=0.025 at IL-6=128 pg/ml and P=0.016 at IL-6=16 pg/ml respectively). IL-6 also inhibited FSH-stimulated but not unstimulated Prog release (P=0.038 at IL-6=8 pg/ml). These findings suggest that IL-6 may be one of the factors that plays a local regulatory role in the course of FSH-stimulated E2 and Prog release. The time-course studies of the effect of the absence or presence of IL-6 demonstrated a significant inhibitory effect on both E2 and Prog secretion (P<0.001) by FSH-stimulated GCs. As infections of the female reproductive tract are often accompanied by elevated local IL-6 levels, this factor may be one of the links leading to endocrine reproductive dysfunction during genital infections. PMID- 11479144 TI - Large body size in the dog is associated with transient GH excess at a young age. AB - The main determinants of body size are GH and IGFs. The aim of this study was to investigate whether differences in adult body size of medium-sized and giant dog breeds can be explained by differences in GH release and/or in plasma IGF-I and IGF-II concentrations at a young age. The basal plasma concentrations of GH, IGF I and IGF-II were determined once weekly in six Great Danes and six beagles from the age of 6 weeks until the age of 24 weeks. In addition, the 6 h secretory profile of GH was determined every 2 weeks. Basal plasma GH concentrations as well as the total area under the curve (AUC) and the AUC above the baseline for GH were significantly higher in Great Danes than in beagles of the same age. In contrast, plasma IGF-I and IGF-II concentrations did not differ significantly between the two breeds. Compared with values in adults, the basal plasma GH concentrations were high until the age of 7 weeks in the beagles, whereas in the Great Danes the basal plasma GH levels remained high during the entire observation period, albeit with a gradual decline. The mean frequency and the mean amplitude of GH pulses tended to be higher in Great Danes than in beagles, although a significant difference was only reached at the age of 19 and 23 weeks for the frequency and at the ages of 9, 11 and 13 weeks for the amplitude. An age dependent decrease in pulse frequency occurred in the Great Danes. The results of this study demonstrate that differences in adult body size of medium-sized and giant dog breeds are preceded by differences in GH release and not by differences in circulating IGF-I or IGF-II concentrations. Both young Great Danes and young beagles experience a period of high GH release, but this period persists much longer in Great Danes. It is discussed that this difference may be due to delayed maturation of the inhibitory influences of somatostatin on pituitary GH release in the latter dogs. PMID- 11479145 TI - Familial resemblance for free androgens and androgen glucuronides in sedentary black and white individuals: the HERITAGE Family Study. Health, Risk Factors, Exercise Training and Genetics. AB - Familial correlation analyses were used to evaluate the familial aggregation of plasma androgens and androgen glucuronides (testosterone (TESTO), dihydrotestosterone (DHT), androstane-3 alpha,17 beta-diol glucuronide (3 alpha DIOL-G), and androsterone glucuronide (ADT-G)) in 505 members of 99 white families and 296 members of 111 black families participating in the Health, Risk Factors, Exercise Training and Genetics (HERITAGE) Family Study. Each of these four measures was determined by RIA after separation of conjugated and unconjugated steroid using C18 column chromatography. All participants were sedentary prior to being including in this study. Significant spouse correlations, as well as parent-offspring and sibling correlations, were found for TESTO, DHT, 3 alpha-DIOL-G, and ADT-G in the white sample, suggesting that common familial environments and genes contribute to the familial resemblance. In the black sample, significant sibling and parent-offspring correlations were found for all four phenotypes, while the spouse correlation was marginally significant for 3 alpha-DIOL-G and not significant for TESTO, DHT, and ADT-G. The non-significance of spouse correlations in the black individuals may be due to the small number of spouse pairs. The maximal heritability estimates of TESTO, DHT, 3 alpha-DIOL-G, and ADT-G were 69%, 87%, 74%, and 56% for white individuals and 70%, 73%, 62%, and 48% for black individuals respectively. Sex differences in heritability estimates were found in the white individuals, but they were less dramatic in the black individuals. In conclusion, plasma levels of androgens and androgen glucuronides are highly heritable in both white individuals and black individuals. There are notable sex differences in the white individuals. PMID- 11479146 TI - Expression of the antiviral protein Mx in peripheral blood mononuclear cells of pregnant and bred, non-pregnant ewes. AB - Interferon-tau (IFN tau) acts locally on the endometrium to suppress estrogen and oxytocin receptor expression and block luteolysis in ruminants. Systemic administration of conceptus homogenates or recombinant ovine IFN tau does not block luteolysis or enhance pregnancy rates in sheep or cattle, respectively. However, IFN tau up-regulates expression of the antiviral protein Mx throughout the entire uterine wall during early pregnancy. These studies determined if conceptus-derived IFN tau also up-regulates Mx expression in components of the circulating immune system that migrate through the endometrial wall. In experiment one, peripheral blood mononuclear cells (PBMC) were isolated from ewes at D26 post-artificial insemination (AI) and Mx mRNA levels examined by Northern and slot-blot hybridization. Pregnancy resulted in a two-fold increase in Mx mRNA levels compared to bred, non-pregnant ewes at D26. In experiment two, PBMC were isolated from ewes at AI, and every three days from D9 to D30. Results showed a four-fold increase in Mx mRNA levels in PBMC from pregnant versus bred, non pregnant ewes at D15. Increased Mx mRNA, which remained elevated through D30, was accompanied by increased levels of Mx protein. These results show that pregnancy recognition signaling rapidly induces Mx gene expression in PBMC, and are the first to suggest that IFN tau activates gene expression in components of the circulating immune system. PMID- 11479147 TI - Daily and nocturnal hemodialysis: how do they stack up? AB - A growing number of articles in the literature describe experiences using more frequent hemodialysis (HD), either short daily or long nocturnal. Most of these publications highlight successes obtained by these programs with a fragmented look at specific areas and outcomes. This review of published results from the use of these therapies shows that universal improvement is noted in dialysis adequacy, nutrition, quality of life, blood pressure control, fluid and electrolyte balance, and hospitalizations when these parameters are mentioned. However, data reporting is often incomplete. Most studies do not have adequate control groups, patient populations are often different from the standard HD population, and many have small numbers that preclude statistical significance. Nonuniformity of patient selection and study design prevents accurate comparison and pooling of patient data. In some cases, the same patients' data for the same periods of observation are reported in several studies. Despite data that can be characterized as preliminary and anecdotal, the results reported in this review show remarkable patient improvement worthy of serious consideration by the renal community. To reach a level of evidence that will be widely acceptable, the renal community needs to partner with such government institutions as the National Institutes of Health and the Health Care Financing Administration to study systematically the outcomes and costs associated with using more frequent HD. In the process, important ramifications of such a cooperative study, including potential changes in policy, need to be considered. PMID- 11479148 TI - Urinary excretion of IgG and alpha(1)-microglobulin predicts clinical course better than extent of proteinuria in membranous nephropathy. AB - In idiopathic membranous nephropathy (MN), the main predictors for progression to chronic renal failure (CRF) are the amount of proteinuria and extent of tubulointerstitial damage. The aim of this study is to evaluate whether urinary excretion of proteins reflecting the alteration of permselectivity in the glomerular capillary wall, such as immunoglobulin G (IgG), and the reabsorption impairment of low-molecular-weight proteins, such as alpha(1)-microglobulin (alpha(1)m), correlates with the extent of tubulointerstitial damage and have a predictive value for functional outcome and response to therapy better than 24 hour proteinuria. In 78 patients with MN, urinary excretion of albumin, transferrin, IgG, and alpha(1)m was measured by immunonephelometry in second morning urine samples and expressed in milligrams per gram of urinary creatinine (uCr). In 48 patients with characterization of proteinuria and renal biopsy performed at the same time, excretion of IgG (P = 0.0087) and alpha(1)m (P = 0.0024), but not albumin (P = 0.37), transferrin (P = 0.38), or 24-hour proteinuria (P = 0.32), was associated significantly with the extent of tubulointerstitial damage (score, 0 to 1 versus >/=2). Only alpha(1)m excretion was associated significantly with global glomerular sclerosis (P = 0.0032) and arteriolar hyalinosis (P = 0.0004). Moreover, urinary excretion of alpha(1)m was significantly dependent on IgG excretion (r = 0.67; P = 0.0001), but not on albumin (P = 0.66) or 24-hour proteinuria (P = 0.07). Functional outcome could be evaluated in 38 patients with nephrotic syndrome and baseline normal renal function (serum creatinine, 0.99 +/- 0.20 mg/dL; follow-up, 44 +/- 22 months). Remission was 100% versus 20% in patients with IgG excretion less than 110 mg/g uCr versus 110 mg/g uCr or greater (P = 0.0001) and 77% versus 17% in patients with alpha(1)m excretion less than 33.5 mg/g uCr versus 33.5 mg/g uCr or greater (P = 0.0009), respectively. In patients with IgG and alpha(1)m excretion less than or greater than the cutoff value, progression to CRF was 0% versus 35% (P = 0.0026) and 0% versus 58% (P = 0.0001), respectively. Nineteen patients treated with immunosuppressive therapy were compared with 19 untreated patients. There was no difference in remission or progression between treated and untreated patients when IgG and alpha(1)m excretion were less than the cutoff value. There was a significant difference for progression to CRF between treated and untreated patients when alpha(1)m excretion was greater than the cutoff value (17% versus 100%; P = 0.0076). In conclusion, IgG excretion is associated significantly with the extent of tubulointerstitial damage and alpha(1)m excretion. This observation supports the hypothesis that IgG may be the toxic moiety of proteinuria. Excretion of IgG and alpha(1)m has a significant predictive value for both remission and progression and is useful to identify patients who are at risk for progression and for whom treatment with immunosuppressive therapy is indicated soon after diagnosis. PMID- 11479149 TI - Follow-up of patients with epidemic poststreptococcal glomerulonephritis. AB - In 1998 there was a large outbreak of acute glomerulonephritis (GN) in Nova Serrana, Brazil, caused by group C Streptococcus zooepidemicus and linked to the consumption of contaminated cheese produced with unpasteurized milk. This study describes the follow-up of these patients after a mean of 2 years following the acute episode. Of 134 patients identified in 1998, 69 patients were reexamined and underwent measurements of blood pressure, 24-hour creatinine clearance, microalbuminuria (radioimmunoassay), and urine sediment analysis. Of the original group of 134 patients, 3 patients died in the acute phase and 5 patients (3.7%) required chronic dialysis. Of 69 patients reevaluated, 65 patients (94%) were adults (mean age, 39 +/- 2 [SE] years) and 47 patients (68%) were women. At the follow-up examination, we found arterial hypertension in 42% of subjects (27 of 64 subjects), serum creatinine levels greater than 1.2 mg/dL in 12% (10 of 68 subjects), reduced creatinine clearance (<80 mL/min/1.73 m(2)) in 30% (20 of 67 subjects, 2 of them on chronic dialysis therapy), and increased microalbuminuria (>20 microg/min) in 34% (22 of 65 subjects). Increased microalbuminuria and/or reduced creatinine clearance were detected in 48% of the subjects (31 of 65 subjects). Patients with microalbuminuria had greater diastolic blood pressure than those without microalbuminuria (mean, 98 +/- 4 versus 88 +/- 2 mm Hg; P = 0.02). In conclusion, after a mean of 2 years, patients with epidemic poststreptococcal GN caused by S zooepidemicus present a high rate of hypertension and frequent abnormalities of renal function, with some having reached end-stage renal disease. Longer follow-up will be important to define the prognosis of these patients. PMID- 11479150 TI - Treatment of diffuse proliferative lupus glomerulonephritis: a comparison of two cyclophosphamide-containing regimens. AB - Cyclophosphamide (CYC) has proven beneficial in preserving renal function in patients with lupus with diffuse proliferative glomerulonephritis (DPGN). However, the optimal route of CYC administration is unknown because direct comparative studies are unavailable. In this open study, we compared the renal outcome of two historical cohorts of patients with diffuse proliferative lupus nephritis (World Health Organization classes IVa and IVb) treated with either intravenous (IV) pulse CYC (group A; n = 22) or sequential oral CYC followed by azathioprine (AZA; group B; n = 21) and followed up prospectively. Both groups of patients had similar clinical, biochemical, and renal parameters at baseline. At 24 months posttreatment, significant improvements in proteinuria, creatinine clearance, serum albumin level, and lupus serological results were evident in both groups. Compared with patients in group A, patients in group B had more complete or partial remission (90% versus 73%) and less risk for treatment failure (5% versus 14%), renal flares (5% versus 14%), and doubling of creatinine levels (5% versus 9%), but the difference was not statistically significant. However, patients treated with oral immunosuppression had an insignificant increase in rates of herpes zoster infection (19% versus 9%) and menstrual disturbance (50% versus 29%). We conclude that sequential oral immunosuppression with CYC and AZA tended to have better efficacy than IV pulse CYC in the treatment of lupus DPGN but was associated with more toxicities. Additional randomized trials involving a larger cohort of patients with a longer period of observation are necessary. PMID- 11479151 TI - Effects of a new synthetic selectin blocker in an acute rat thrombotic glomerulonephritis. AB - In an attempt to explore a novel therapeutic approach, a new synthetic sulfatide derivative (SKK60037) was evaluated in an acute rat model of P-selectin and leukocyte-dependent thrombotic glomerulonephritis (TG). In vitro, SKK60037 inhibits the function of P- and L-selectin more effectively than sialyl Lewis X (sLe(x)), a well-established selectin blocker. TG was induced by the intravenous administration of nephrotoxic globulin (NTG) to rats pretreated with a subclinical dose of lipopolysaccharide. In this model, platelet accumulation was remarkable within 10 minutes after induction of disease, followed by the infiltration of leukocytes, mainly neutrophils and macrophages. Thrombus formation and fibrinogen deposition in the glomeruli were observed within 1 hour, and they proceeded until 6 hours. P-selectin was highly expressed in glomeruli, whereas E-selectin and L-selectin ligands were not detected. We tested the effects of SKK60037 in this model in comparison with sLe(x) and antirat P selectin monoclonal antibody (ARP2-4). SKK60037 blocked platelet accumulation in glomerular capillaries at 10 minutes after NTG injection. At 6 hours, leukocyte infiltration and thrombosis were significantly suppressed. Protective effects of SKK60037 were similar to those of ARP2-4, whereas sLe(x) showed minimum effect. The superior effects and more favorable characteristics of SKK60037 to sLe(x) suggest the potential of SKK60037 for clinical application. PMID- 11479152 TI - Hepatocyte growth factor inhibits apoptosis after ischemic renal injury in rats. AB - Several studies have determined that growth factors, including hepatocyte growth factor (HGF), have a crucial role in the regenerative process of renal tubules after ischemic or toxic insult. Recent research has ascertained that as well as necrotic cell death, there is evidence of apoptosis after an acute renal injury. We attempted to determine the effect of HGF on apoptosis after ischemic renal injury in rats. We administered HGF or vehicle to 12 rats after ischemic insult and compared them with 6 sham-operated controls. Rats were killed at 48 hours, and histopathologic assessments were performed on the renal tissue. The microscale autoradiographic method was used for qualitative analysis of DNA fragmentation. This method was chosen over the widely used ethidium bromide staining method because it increases the sensitivity of detection of apoptotic DNA. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling histopathologic staining was used to identify apoptosis in situ. Apoptotic changes were clearly shown by electron microscopy in vehicle treated animals. Despite showing profound evidence of tubular necrosis, apoptotic changes were markedly reduced in HGF-treated animals compared with vehicle treated animals. DNA-laddering analysis further confirmed the antiapoptotic effect of HGF. To our knowledge, this is the first in vivo illustration of the inhibitory activity of a growth factor on apoptosis in the setting of tubular necrosis. The role of apoptosis in the setting of acute renal failure has not been elucidated; thus, additional research is necessary to determine the significance of these findings. PMID- 11479153 TI - Nephromegaly and elevated plasma hepatocyte growth factor-transforming growth factor-beta1 ratio in infants with fulminant hepatitis or biliary atresia. AB - Nephromegaly, assessed by calculating kidney volume using renal ultrasound, was studied in infants with biliary atresia, neonatal hepatitis, or fulminant hepatitis. We evaluated kidney volume in 29 patients with biliary atresia, 17 patients with neonatal hepatitis, and 10 patients with fulminant hepatitis, as well as 32 healthy infants. Levels of plasma hepatocyte growth factor (HGF) were measured in all infants. Levels of plasma transforming growth factor-beta1 (TGF beta1) were also measured in diseased infants and 20 healthy infants. Significant nephromegaly was found in infants with biliary atresia compared with healthy infants (P < 0.001 by analysis of covariance). Marked nephromegaly was also noted in all infants with fulminant hepatitis and 35% of infants with neonatal hepatitis. No nephromegaly was found in infants at 2 months of age with biliary atresia or neonatal hepatitis despite mildly elevated plasma HGF levels. Regardless of the duration of HGF exposure and healthy renal growth by a certain age, a positive correlation existed between plasma HGF level and kidney volume (r = 0.529; P < 0.001), but an inverse correlation was found between plasma TGF beta1 level and nephromegaly (r = -0.505; P < 0.001) in all diseased infants. There was a stronger positive correlation between plasma HGF-TGF-beta1 ratio and kidney volume (r = 0.666; P < 0.001) and degree of nephromegaly (r = 0.717; P < 0.001). These results confirm the presence of large kidneys not only in patients with biliary atresia but also in patients with fulminant hepatitis, which suggests the possible pathogenic role of HGF and manifests as elevated HGF-TGF beta1 ratios in patients with such conditions. Nephromegaly in patients with severe or chronic liver dysfunction may provide a new in vivo model to study the mechanisms of renal growth. PMID- 11479154 TI - Polarization of T-helper lymphocytes toward the Th2 phenotype in uremic patients. AB - T-helper (Th) lymphocytes consist of Th1 and Th2 subsets. Th1 cells are effectors of cell-mediated immunity and secrete interferon-gamma (IFN-gamma), which recruits new Th1 cells in cooperation with interleukin-12 (IL-12; produced by monocytes) and inhibits Th2 differentiation. Th2 cells produce IL-4 and IL-10, which inhibit IFN-gamma secretion and cell immunity. We investigated whether the impaired immune response in uremia is associated with an altered balance of Th1/Th2. Peripheral-blood mononuclear cells (PBMCs) were collected from patients with chronic renal failure (CRF) on conservative treatment (CRF patients), patients with end-stage renal disease (ESRD) on regular hemodialysis therapy (ESRD-HD patients), and healthy controls (CON). CD4(+) cells were isolated from PBMCs by negative selection using a magnetic labeling system. PBMCs and purified CD4(+) cells were cultured in Iscove's medium and Iscove's medium plus mitogens (phytohemagglutinin and lipopolysaccharide). IFN-gamma, IL-12, IL-4, and IL-10 were measured in supernatant. The constitutive release of IL-4 and IL-10 by PBMCs and CD4(+) cells of CRF and ESRD-HD patients was increased by five to eight times in comparison with CON (P < 0.001). Constitutive IFN-gamma release by PBMCs of ESRD-HD patients was undetectable, although they secreted an increased amount of IL-12. Mitogen-stimulated release of IFN-gamma by PBMCs and CD4(+) cells of CRF and ESRD-HD patients was blunted (average PBMCs: CON, 115.8 pg/2 x10(6) cells; CRF, 81.8 pg/2 x10(6) cells; ESRD-HD, 9.3 pg/2 x10(6) cells; CD4(+) cells: CON, 358.0 pg/5 x 10(5) cells; CRF, 165.4 pg/5 x 10(5) cells; ESRD-HD, 43.5 pg/5 x 10(5) cells). The ability of PBMCs of ESRD-HD patients to secrete IFN-gamma was recovered after IL-4 and IL-10 neutralization. Uremia is associated with a prevalence of Th1 over Th2 cells and a configuration of cytokine network that depresses cell-mediated immunity. PMID- 11479155 TI - Comet-assay analysis identifies genomic damage in lymphocytes of uremic patients. AB - This study investigates genomic damage in peripheral lymphocytes from patients with moderate to severe chronic renal insufficiency and those on long-term maintenance hemodialysis (MHD) and hemodiafiltration therapy. As a measure for genomic damage, the comet assay (single-cell gel electrophoresis) was applied. This test detects single- and double-strand breaks and alkali labile sites through electrophoretic mobility of the resulting fragments. The average damage (percentage of DNA in the tail region of the comet) observed in cells of the control group of 21 healthy subjects was 10.5% +/- 0.8%. There was a significant increase to 14.7% +/- 3.5% in cells of 23 patients with chronic renal failure, and a further increase to 17.1% +/- 3.5% in the subgroup of 12 patients with serum creatinine values greater than 6 mg/dL. Damage was 16.7% +/- 4.2% in cells of the MHD group (26 patients) and 20.1% +/- 3.0% in the subgroup with MHD therapy longer than 10 years (8 patients). Cellular DNA damage in the group of 15 maintenance hemodiafiltration patients was 15.6% +/- 2.1%, ranging between predialysis and MHD patients, and did not seem to increase with treatment time. These results, together with previously observed elevated frequencies of micronuclei, decreased DNA repair, and increased cancer incidence described for these patient groups, emphasize the need to further optimize the current therapy for reducing the degree of genomic damage. PMID- 11479156 TI - Presentation and revascularization outcomes in patients with radiation-induced renal artery stenosis. AB - This study analyzed the initial presentation and revascularization outcomes of patients with radiation-induced renal artery stenosis, a rare complication of therapeutic irradiation. Of 11 patients with renal artery stenosis after irradiation, 7 patients fulfilled the following criteria: normotension before irradiation, radiation dose greater than 25 grays delivered to the renal arteries, associated perirenal radiation-induced lesions, and absence of arterial disease outside the radiation field. The median age at irradiation was 30 years, and the median local irradiation dose was 40 grays. The median time from irradiation to referral was 13 years. All patients were hypertensive at referral, with a median blood pressure (BP) of 171/102 mm Hg and median treatment score of two. The median glomerular filtration rate was 67 mL/min. Two patients had bilateral stenoses and 1 patient had stenosis affecting a single kidney. Stenoses were proximal in 6 patients and truncal in 1 patient, and all had the appearance of atherosclerotic stenosis. Percutaneous transluminal renal artery angioplasty (PTRA) was successful in 5 patients, but required multiple insufflations. PTRA failed in 1 patient, who subsequently underwent an aortorenal bypass. After a median follow-up of 36 months, 2 patients had died of noncardiovascular causes and 4 patients remained hypertensive, with a median BP of 136/85 mm Hg and median treatment score of two. No restenosis occurred, but aneurysms developed at the site of angioplasty in 1 patient. If hypertension occurs even decades after irradiation, a radiation-induced renal artery stenosis should be sought in patients who have undergone abdominal irradiation. PMID- 11479157 TI - Cystatin C is not more sensitive than creatinine for detecting early renal impairment in patients with diabetes. AB - This study evaluated serum cystatin C as a potential new marker of glomerular filtration rate (GFR) in 49 patients who had steady-state diabetes with early renal impairment. We determined the correlation between GFR measured by chromium 51-labeled EDTA and levels of serum cystatin C, serum creatinine, serum beta(2) microglobulin, endogenous creatinine clearance, and Cockcroft formula. Sensitivity and specificity for the diagnosis of renal failure, defined as a GFR less than either 80 or 60 mL/min/1.73 m(2), were calculated by receiver operating characteristic (ROC) curves for creatinine, cystatin C, and beta(2) microglobulin. Finally, we compared mean values of these three serum parameters in patients grouped according to GFR using the two definitions of renal failure. Correlation coefficients with GFR were -0.77 for serum creatinine level, -0.65 for serum cystatin C level, -0.71 for serum beta(2)-microglobulin level, +0.56 for endogenous creatinine clearance, and +0.69 for Cockcroft formula (all P < 0.001). With a cutoff value of 60 mL/min/1.73 m(2), areas under the ROC curve were 0.972 for beta(2)-microglobulin, 0.925 for cystatin C, and 0.916 for creatinine levels. With a cutoff value of 80 mL/min/1.73 m(2), these were 0.838 for beta(2)-microglobulin, 0.780 for cystatin C, and 0.905 for creatinine levels (P = not significant between parameters). These results were not altered after the exclusion of patients (n = 8) with a serum creatinine level greater than 1.41 mg/dL. When patients were classified into three groups according to GFR (group 1, >80 mL/min/1.73 m(2); group 2, 60 to 80 mL/min/1.73 m(2); group 3, <60 mL/min/1.73 m(2)), mean values of serum parameters in the three groups were statistically different (P < 0.0001) except between groups 1 and 2 for cystatin C and beta(2)-microglobulin. With patients classified into two groups (GFR > or < 80 mL/min/1.73 m(2)), mean values for each parameter were statistically different (P < 0.001). Sensitivity, specificity, and positive and negative predictive values for serum creatinine and serum cystatin C levels were very close for both definitions of renal failure. Serum cystatin C is not better than serum creatinine or serum beta(2)-microglobulin levels for estimating GFR in patients with steady-state diabetes using ROC curves or other validation tests. PMID- 11479158 TI - In vitro studies of aquaporins 1 and 3 expression in cultured human proximal tubular cells: upregulation by transferrin but not albumin. AB - Dysregulated renal water handling is a cardinal feature of nephrotic syndrome that has been shown in animal models of experimental nephrosis to mediate renal aquaporin (AQP) expression. However, data on the effect of proteinuria on the proximal tubule, which is heavily vested with AQP1 and therefore may participate in water homeostasis, are limited. To investigate this, we exposed primary human proximal tubular epithelial cells (PTECs) to two key proteinuric components shown to perturb tubule function: human serum albumin and transferrin. Using reverse transcriptase polymerase chain reaction and immunocytochemical techniques, PTECs in the quiescent state were found to express AQP3 in addition to AQP1 gene and protein, which was also validated in a human proximal tubule cell line, HK-2. Immunohistochemical staining localized AQP1 synthesis to the apical and basolateral membranes and AQP3 synthesis to the basolateral membrane of proximal tubule epithelium. Transferrin in doses reaching nephrotic range upregulated PTEC transcription and translation of both AQP1 and AQP3 in a time- and dose-dependent manner. After 24 hours of stimulation, transferrin led to a 2.4- and 2.2-fold increase in AQP1 and APQ3 messenger RNA expression, whereas protein synthesis surged by 40.7% +/- 2.48% and 24.2% +/- 0.9% compared with control, respectively. These effects were not observed with albumin challenge and were not caused by osmolality fluctuation with transferrin treatment. In summary, our novel finding of AQP3 in PTECs indicates a role for AQP3 in proximal tubule water reabsorption. The pathophysiological significance of heightened AQP1 and AQP3 expression in PTECs on protein challenge as occurs in the nephrotic state requires further investigation. PMID- 11479159 TI - Cytokine production induced by binding and processing of calcium oxalate crystals in cultured macrophages. AB - Deposition of calcium oxalate (CaOx) crystals in the renal interstitium is common in humans with primary oxalosis and secondary hyperoxaluria, as well as in kidneys of rats with CaOx nephrolithiasis. In vivo, macrophages and multinucleated giant cells mostly encapsulate these crystals. To investigate whether macrophages are able to dispose of CaOx crystals after phagocytosis, we used a nontransformed macrophage cell line derived from mouse spleen progenitors. Cytokine assays showed that in response to crystal binding and phagocytosis, these macrophages release tumor necrosis factor-alpha. This release was evident at 8 hours, maximal at 24 hours, and decreased to control values after 48 hours of incubation with crystals. A very low but significant release of interleukin-6 into the culture medium was only noticed after 32 hours. Radiochemical experiments showed that these cells bind 38.8% of the CaOx crystals added. After 4 days, all internalized crystals had been dissolved and their molecular constituents released into the extracellular environment. Confocal laser scanning microscopy followed by morphometrical analyses confirmed these results. Long-term (survival) analyses showed that in the interval under study and at the crystal doses used, cell viability was not significantly affected. These findings support the view that properly functioning macrophages are able to remove CaOx deposits from the renal interstitium and that these cells produce inflammatory cytokines before crystal dissolution. PMID- 11479160 TI - Ethylene glycol-mediated tubular injury: identification of critical metabolites and injury pathways. AB - Ethylene glycol (EG) intoxication produces multisystem organ injury, including acute renal failure. Although EG must be metabolized to toxic intermediates to induce organ damage, the specific metabolite(s) responsible and the underlying pathogenic mechanisms remain poorly defined. To explore these issues, isolated mouse proximal tubular segments (PTSs) were incubated with either varying doses of EG or its prime metabolites (glycolate, glycoaldehyde, glyoxylate, or oxalate for 15 to 60 minutes). Injury was assessed by the percentage of lactate dehydrogenase (LDH) release, LDH destruction, adenosine triphosphate (ATP) depletion, or membrane phospholipid degradation. Toxicities were also assessed in cultured HK-2 cells over 18 hours (by MTT assay). EG, glycolate, and oxalate did not induce overt PTS injury. Conversely, glyoxylate and glycoaldehyde were highly toxic, causing profound ATP depletion and LDH release. Glycoaldehyde also caused enzyme (LDH) and selected phospholipid degradation (phosphatidylethanolamine, phosphatidylserine). These changes were not seen with glyoxylate treatment. Acidosis (pH 6.8) and glycine (2 mmol/L) each blocked glyoxylate, but not glycoaldehyde toxicity, indicating differing injury pathways. Only glycoaldehyde and glyoxylate induced marked HK-2 cell death. We conclude that glycoaldehyde and glyoxylate are the principal metabolites responsible for EG nephrotoxicity and do so by causing ATP depletion and phospholipid and enzyme destruction. Glycine and acidosis, by-products of EG metabolism, can attenuate glyoxylate-mediated injury. This suggests that naturally occurring but incomplete protective pathways may be operative during the evolution of EG cytotoxicity. PMID- 11479161 TI - Variables affecting weight gain in renal transplant recipients. AB - Previous studies of renal transplant recipients have suggested that weight gain after transplantation is relatively common, especially among certain populations. We conducted a retrospective review of 977 renal transplant recipients at the University of Alabama at Birmingham to identify patterns of weight change (as mean percentage of body weight at transplantation) attributed to race, sex, income, age at transplantation, pretransplantation time on dialysis, incidence of diabetes, rejection episode(s), and/or obesity (body mass index >/= 30 kg/m(2)) at transplantation. Patients were evaluated at 3, 6, 9, and 12 months posttransplantation and at 2 and 3 years, when available. Univariate analysis at 1 year showed that blacks achieved a greater weight change than whites (P = 0.0004), women had greater gains than men (P = 0.0001), and low-income patients had greater mean gains versus medium- (P = 0.0001) and high-income patients (P = 0.0001). Advancing age and weight gain were inversely correlated (P = 0.0002). Having one or more rejection episode indicated less weight gain than having no rejection episode (P = 0.0220). Incidence of diabetes or time on dialysis was not a significant predictor of weight gain. Black race, female sex, low income, younger age, and no incidence of rejection episodes were significantly associated with weight gain at 1 year in the multivariate analysis. PMID- 11479162 TI - Molecular genotyping of BK and JC viruses in human polyomavirus-associated interstitial nephritis after renal transplantation. AB - The human polyomaviruses BK virus (BKV) and JC virus (JCV) have been linked to ureteric stenosis and allograft interstitial nephritis, but molecular characterization of the species involved has not been performed. We studied paraffin-embedded renal tissue from 19 cases of allograft viral interstitial nephritis. Histological sections were subjected to polymerase chain reaction amplification using consensus, BKV-, and JCV-specific primers, with subsequent DNA sequencing for strain determination. BKV was present in all (100%) interstitial nephritis kidneys and placed in genotypes corresponding to serological groups I (n = 11), II (n = 1), and IV (n = 5). Fourteen of 17 isolates (82%) showed sequence variations in the viral capsid protein-1 (VP1) capsid region, with predicted changes in the encoded amino acids and sometimes with potential implications for the secondary and tertiary structure of the corresponding protein molecules. An additional case showed a previously reported glutamine-->leucine T-antigen region mutation. JCV was seen in seven interstitial nephritis kidneys (37%), with types 4 (n = 3), 3A (n = 2), and 2A (n = 1) identified. Most white individuals with asymptomatic infection are reported to shed type 1 JCV in the urine. Simian 40 polyomavirus was not identified in any case. These observations may have pathogenic relevance to the development of an extremely refractory form of polyomavirus interstitial nephritis seen after kidney transplantation. PMID- 11479163 TI - Body composition in children after renal transplantation. AB - This study examines changes in body composition after renal transplantation (Tx) in 16 children who underwent dual-energy X-ray absorptiometry examination within 6 months preceding Tx; the examination was then repeated every 6 to 12 months after Tx. Body composition was analyzed with regard to whole-body bone mineral content (BMC), lean body mass (LBM), and fat body mass (FBM). Median BMC decreased from the initial value of +0.98 standard deviation scores (SDS) to 0.55 SDS during the first 3 months after Tx, and a further decrease was noted at the end of month 6 (M6; -1.34 SDS) and M12 after Tx (-1.32 SDS). Improvement was observed during the second year after Tx (P for global changes < 0.0001). LBM and FBM also changed significantly during the 2 years after Tx (P = 0.006 and P = 0.0001, respectively). LBM decreased during the first 3 months after Tx (median change, 0.71 SDS) and remained less than 0 SDS in all but 4 patients. Median LBM did not decrease to less than -1 SDS during the entire study period. Conversely, median FBM increased by a median of 3.73 SDS during the first 3 months and remained elevated during the first 12 months after Tx, with a subsequent decrease at 2 years after Tx. No significant correlation was found between cumulative doses of prednisone and BMC, FBM, or LBM at any interval if absolute values were considered. However, relative changes in FBM correlated significantly with relative changes in prednisone cumulative doses. PMID- 11479164 TI - Short daily hemodialysis: blood pressure control and left ventricular mass reduction in hypertensive hemodialysis patients. AB - Several retrospective and uncontrolled prospective studies reported blood pressure (BP) normalization and left ventricular mass (LVM) reduction during daily hemodialysis (DHD). Conversely, the burden of these major independent risk factors is only marginally reduced by the initiation of standard thrice-weekly dialysis (SHD), and cardiovascular events still represent the most common cause of death in hemodialysis patients. Therefore, we performed a randomized two period crossover study to compare the effect of short DHD versus SHD on BP and LVM in hypertensive patients with end-stage renal disease. We studied 12 hypertensive patients who had been stable on SHD treatment for more than 6 months. At the end of 6 months of SHD and 6 months of DHD in a sequence of randomly assigned 24-hour ambulatory BP monitoring, echocardiography and bioimpedance were performed. Throughout the study, patients maintained the same Kt/V. A significant reduction in 24-hour BP during DHD was reported (systolic BP [SBP]: DHD, 128 +/- 11.6 mm Hg; SHD, 148 +/- 19.2 mm Hg; P < 0.01; diastolic BP: DHD, 67 +/- 8.3 mm Hg; SHD, 73 +/- 5.4 mm Hg; P = 0.01). The decrease in BP was accompanied by the withdrawal of antihypertensive therapy in 7 of 8 patients during DHD (P < 0.01). LVM index (LVMI) decreased significantly during DHD (DHD, 120.1 +/- 60.4 g/m(2); SHD, 148.7 +/- 59.7 g/m(2); P = 0.01). Extracellular water (ECW) content decreased from 52.7% +/- 11.4% to 47.6% +/- 7.5% (P = 0.02) and correlated with 24-hour SBP (r = 0.63; P < 0.01) and LVMI (r = 0.66; P < 0.01). In conclusion, this prospective crossover study confirms that DHD allows optimal control of BP, reduction in LVMI, and withdrawal of antihypertensive treatment. These effects seem to be related to reduction in ECW content. PMID- 11479165 TI - Evidence of calcium phosphate depositions in stenotic arteriovenous fistulas. AB - This study investigates vascular samples from patients with and without end-stage renal disease (ESRD) to determine the occurrence of calcium depositions. Findings in stenotic arteriovenous (AV)-fistula veins were compared with those of nonstenotic AV-fistula veins, non-AV-fistula veins, and atherosclerotic vessels. Calcium and phosphorus content was measured by means of scanning electron microscopy and its built-in method of energy-dispersive spectrometry (EDS) X-ray analysis. We found calcium and phosphorus in samples from AV fistulas with stenotic areas with a calcium/phosphorus molar ratio of 1. Based on EDS analysis and crystal shape comparison, we conclude that calcium phosphate precipitations in stenotic AV fistulas are brushites with the composition CaHPO(4)*2H(2)O. This specific calcium phosphate deposition was found solely in stenotic AV fistulas, not in nonstenotic AV-fistula veins or non-AV-fistula veins regardless of whether the patient had ESRD. Moreover, this calcium phosphate deposition was different from calcium compounds found in atherosclerotic samples. Whether the precipitation of brushite is primarily involved in the development of vascular access stenosis or represents a secondary consequence cannot be determined from the present study. PMID- 11479166 TI - Cellulose membranes suppress complement activation in patients after hemodialysis. AB - Membranes used for dialysis therapy activate complement. Complement activation is maximal after initiating dialysis and returns to predialysis values by the end of dialysis. No changes in C3 levels have been detected after dialysis. We hypothesized that although C3 levels were unchanged, C3 activity could be altered by dialysis. We measured complement activation in vitro in serum from patients randomized to dialysis treatments using different types of membranes. The classical pathway was activated with aggregated immunoglobulin G (IgG), and the alternative pathway was activated with inulin. Both the classical and alternative pathways were suppressed after dialysis using cellulose membranes (aggregate IgG, P < 0.01; inulin, P < 0.001). When polyacrylonitrile (PAN) or polyethylene glycol grafted cellulose membranes were used for dialysis, only minor suppression of complement pathways was measured. Levels of the control factor SP-40,40 increased at later times for dialysis using cellulose membranes (P < 0.05). Factor H levels were also greater after dialysis using cellulose membranes compared with PAN membranes (P < 0.05). In summary, cellulose membranes suppress complement activation in serum. One suppressing factor may be the complement control factor SP-40,40. PMID- 11479167 TI - Hypokalemic metabolic acidosis attributed to cough mixture abuse. AB - This report describes a patient with mixed normal anion gap hyperchloremic metabolic and respiratory acidosis associated with hypokalemia attributed to cough mixture abuse. Metabolic acidosis was likely related to an overdose of ammonium chloride, whereas respiratory acidosis was probably related to the effect of hypokalemia on respiratory muscles, causing hypoventilation. Hypokalemia was caused by a transcellular shift of potassium induced by ephedrine and pseudoephedrine. Both ammonium chloride and ephedrine were probably present in the cough mixture obtained by our patient as an over-the-counter medication. Physicians should be aware of the potential for cough mixture abuse to cause major electrolyte disturbances that may carry the risk for major cardiac arrhythmias, particularly in youth. PMID- 11479168 TI - Immunotactoid glomerulopathy with massive bone marrow deposits in a patient with IgM kappa monoclonal gammopathy and hypocomplementemia. AB - A case of immunotactoid glomerulopathy with an amyloid-like material in the glomeruli and bone marrow is described. Clinically the patient was diagnosed as having severe nephrotic syndrome, hypocomplementemia, and IgM kappa monoclonal gammopathy. Immunotactoid glomerulopathy is an unusual cause of glomerulonephritis, characterized by Congo red-negative, amyloid-like deposits in the glomeruli. This unusual case presentation shows that immunotactoid glomerulopathy may be a manifestation of systemic disease. This patient also presented with hypocomplementemia, an extremely rare associated finding that has been reported previously in only four cases of immunotactoid glomerulopathy. PMID- 11479169 TI - Nonconvulsive status epilepticus in peritoneal dialysis patients. AB - This report describes nonconvulsive status epilepticus (NCSE) in four patients with end-stage renal disease (ESRD) on peritoneal dialysis therapy. All patients presented with acute confusion or stuporous state without clinical motor seizures. Three patients had active systemic infections and were being administered antibiotics. Diagnoses were confirmed in all cases by electroencephalogram (EEG), which showed characteristic diffuse sharp waves and spikes. The EEG appearance improved after anticonvulsant therapy. All patients had satisfactory control of their electrical seizure activity and made varying degrees of mental recovery. To our knowledge, this is the first reported series of NCSE in patients with ESRD on peritoneal dialysis therapy. It shows NCSE as a differential diagnosis for acute confusion. We emphasize the need for nephrologists to consider NCSE as a differential diagnosis for uremic encephalopathy. PMID- 11479170 TI - The more things change... PMID- 11479171 TI - Weight gain after kidney transplantation. PMID- 11479172 TI - Targets and targeting. AB - Using the vocabulary of ballistics in medicine for emphasis can result in misleading exaggeration and semantic confusion. The dual meaning of target as either aiming point (aim at) or outcome (aim to achieve) creates a muddle in the efforts to comply with quality assurance initiatives. Disentangling the two meanings allows new approaches to the clinical technology required in a modern health care environment. An example can be shown in new strategies for the management of renal anemia with iron and erythropoietin. The potential to shape outcome distributions through validated, preemptive intervention thresholds offers the predictable results required by patients and payers. Using the management of patient cohorts as a platform for outcomes creates no necessary conflict with individualized clinical care. Future guideline statements should include the likely characteristics of compliant outcome populations, as a prompt to clinical goals and as an indication of the necessary cost and effort of compliance with treatment standards. Overemphasis in language is no substitute for considered clinical methodology. PMID- 11479173 TI - Why is erythropoietin made in the kidney? The kidney functions as a critmeter. AB - Erythropoietin is distinct among the hematopoietic growth factors because it is produced primarily in the kidneys rather than the bone marrow. The kidney functions as a critmeter in that it senses oxygen tension and extracellular volume. By regulating red cell mass through erythropoietin and plasma volume through excretion of salt and water, the kidney sets the hematocrit at a normal value of 45%. This is not a random number, but a value that maximizes oxygen delivery to peripheral tissues. The ability of the kidney to coordinate these two volumes to generate a hematocrit of 45% establishes it as the logical site for erythropoietin production. The kidney has the unique ability to translate a measure of plasma volume as tissue oxygen pressure required to regulate erythropoietin production. I hypothesize that the critmeter is a functional unit that regulates the hematocrit. The critmeter is found at the tip of the juxtamedullary region of the cortical labyrinth in the kidney, where erythropoietin is made physiologically. Renal vasculature and nephron segment heterogeneity in sodium reabsorption generate the marginal tissue oxygen pressure required to trigger the production of erythropoietin. The balance of the oxygen consumption for sodium reabsorption and the oxygen delivery to the proximal tubule is reflected by the tissue oxygen pressure that determines red blood cell mass adjusted to plasma volume. Factors that affect blood supply and sodium reabsorption in a discordant manner may modulate the critmeter (eg, angiotensin II). Examples of clinical disorders caused by dysfunction or resetting of the critmeter are described. PMID- 11479174 TI - Role of the nephrologist in the intensive care unit. AB - Intensive care units (ICUs) are increasingly becoming a focal point for tension between medical specialists. In an extreme approach to this issue, some ICUs have become closed units managed by intensivists, with other specialists, such as nephrologists, having a restricted supportive role. The nephrologist, a subspecialist with broad skills in general internal medicine, has trained and appropriately can serve as the primary physician for patients with significant renal failure and end-stage renal disease in multiple hospital settings, including the ICU. Sick and complex hospitalized patients offer ample opportunity for a collaborative interaction between the nephrologist and intensivist in the ICU. PMID- 11479175 TI - Complement and renal ischemia-reperfusion injury. PMID- 11479176 TI - A 76-year-old woman with nephrotic syndrome. PMID- 11479177 TI - Uric acid or urate nephropathy? PMID- 11479180 TI - Chronic subdural hematoma in a 50-year-old man with polycystic kidney disease. AB - The following case describes two spontaneous chronic subdural hematomas in one patient. The hematomas occurred on opposite sides. The patient had good blood pressure control and a normal creatinine. He was not receiving any anticoagulants or antiplatelet drugs. An angiogram was done after the second subdural collection was drained. There was no evidence of a vascular lesion. The possibility remains that patients with adult polycystic disease may be predisposed to develop spontaneous subdural hematomas in light of another report of five other cases. PMID- 11479181 TI - Granulomatous renal disease in a patient with common variable immunodeficiency. AB - Common variable immunodeficiency (CVID), the most common cause of primary hypogammaglobulinemia, is characterized by a decreased serum immunoglobulin level, recurrent infections, and the occurrence of various autoimmune diseases. Granulomatous disease has been reported previously in several patients with CVID, with granuloma occurring in the lymph nodes, spleen, liver, central nervous system, and bone marrow. We report the first published case of renal granulomatous disease in a CVID patient presenting with subacute renal failure. Renal function partially recovered after corticosteroid treatment and intravenous immunoglobulin infusions. The pathogenesis of granulomatous disease in CVID is unclear but may involve monocyte and T-cell abnormalities. PMID- 11479182 TI - Hemodialysis in management of hypothermia. AB - Hypothermia is defined as a core body temperature of less than 35 degrees C and is divided further into mild, moderate, and severe depending on the temperature level. Several active internal rewarming modalities have been described in the management of moderate-to-severe accidental hypothermia. We report a 73-year-old black man with underlying end-stage renal failure and ischemic cardiomyopathy who was admitted with severe accidental hypothermia (core body temperature, 24.9 degrees C) secondary to environmental cold exposure. The patient was resuscitated initially with warm intravenous fluids and peritoneal dialysis with warm fluids with an average temperature rise of 1 degrees C. The patient was switched to hemodialysis that brought his temperature from 30.2 degrees C to 36.7 degrees C during a 3.5-hour dialysis with an average rise of 1.9 degrees C/h. Hemodialysis is a rapid and efficient modality of rapid internal rewarming for moderate-to severe accidental hypothermia. PMID- 11479183 TI - HFE gene and hereditary hemochromatosis: a HuGE review. Human Genome Epidemiology. AB - Hereditary hemochromatosis (HHC) is an autosomal recessive disorder of iron metabolism characterized by increased iron absorption and deposition in the liver, pancreas, heart, joints, and pituitary gland. Without treatment, death may occur from cirrhosis, primary liver cancer, diabetes, or cardiomyopathy. In 1996, HFE, the gene for HHC, was mapped on the short arm of chromosome 6 (6p21.3). Two of the 37 allelic variants of HFE described to date (C282Y and H63D) are significantly correlated with HHC. Homozygosity for the C282Y mutation was found in 52-100% of previous studies on clinically diagnosed probands. In this review, 5% of HHC probands were found to be compound heterozygotes (C282Y/H63D), and 1.5% were homozygous for the H63D mutation; 3.6% were C282Y heterozygotes, and 5.2% were H63D heterozygotes. In 7% of cases, C282Y and H63D mutations were not present. In the general population, the frequency of the C282Y/C282Y genotype is 0.4%. C282Y heterozygosity ranges from 9.2% in Europeans to nil in Asian, Indian subcontinent, African/Middle Eastern, and Australasian populations. The H63D carrier frequency is 22% in European populations. Accurate data on the penetrance of the different HFE genotypes are not available. Extrapolating from limited clinical observations in screening studies, an estimated 40--70% of persons with the C282Y homozygous genotype will develop clinical evidence of iron overload. A smaller proportion will die from complications of iron overload. To date, population screening for HHC is not recommended because of uncertainties about optimal screening strategies, optimal care for susceptible persons, laboratory standardization, and the potential for stigmatization or discrimination. PMID- 11479184 TI - Risk of incident age-related eye diseases in people with an affected sibling : The Beaver Dam Eye Study. AB - The purpose of this investigation was to determine whether age-related cataract and maculopathy in older siblings predicts development of the same in younger siblings. A population-based study of age-related eye diseases was conducted in 1988--1990 in Beaver Dam, Wisconsin, and a follow-up examination was performed 5 years later. Diagnoses of age-related eye diseases were assigned on the basis of gradings of study photographs. There were 1,088 people from 488 sibships with at least two siblings who could contribute information for these analyses. The authors computed odds ratios and 95% confidence intervals for developing the specific lesion and identifying it 5 years later if an older sibling had it at baseline. The odds ratios were 1.65 (95% confidence interval (CI): 0.91, 2.99) for nuclear cataract, 1.62 (95% CI: 0.92, 2.85) for cortical cataract, 1.95 (95% CI: 0.48, 7.95) for posterior subcapsular cataract, 1.82 (95% CI: 0.91, 3.66) for soft drusen, 8.18 (95% CI: 3.34, 20.08) for retinal pigment epithelium depigmentation, 3.59 (95% CI: 1.71, 7.57) for increased retinal pigment, and 10.32 (95% CI: 0.83, 128.58) for exudative age-related maculopathy. These findings suggest that strong family determinants of lesions of age-related maculopathy are likely, less so for age-related cataract, which confer risk of the same lesion in a younger sibling. PMID- 11479185 TI - Effect of teenage pregnancy on educational disabilities in kindergarten. AB - Teenage pregnancies have become a public health issue because of their observed negative effects on perinatal outcomes and long-term morbidity. The association of young maternal age and long-term morbidity is usually confounded, however, by the high prevalence of poverty, low level of education, and single marital status among teenage mothers. The authors assess the independent effect of teenage pregnancy on educational disabilities and educational problems in a total population of children who entered kindergarten in Florida in 1992--1994 and investigate how controlling for potentially confounding factors affects the relation between teenage pregnancies and poor outcome. When no other factors are taken into account, children of teenage mothers have significantly higher odds of placement in certain special education classes and significantly higher occurrence of milder education problems, but when maternal education, marital status, poverty level, and race are controlled, the detrimental effects disappear and even some protective effects are observed. Hence, the increased risk for educational problems and disabilities among children of teenage mothers is attributed not to the effect of young age but to the confounding influences of associated sociodemographic factors. In contrast to teen age, older maternal age has an adverse effect on a child's educational outcome regardless of whether other factors are controlled for or not. PMID- 11479186 TI - Mortality of men versus women in comparable high-level jobs: 15-year experience in the Federal Women's Study. AB - The authors investigated exposure to high-level occupations in relation to the well-known survival advantage of women compared with men of the same age. Women in the federal workforce in positions of General Schedule 14 and above in 1979- 1993 (n = 4,727) were each matched with three men (n = 14,181) by age, General Schedule level, and supervisory role. Fifteen-year mortality rates were compared between men and women and against expected 15-year mortality from the US general population. Despite similar job demands, women experienced markedly lower 15-year mortality than did men. However, men in these positions had nearly 50% lower mortality compared with age-matched men in the general population; the comparable reduction for women was 38%. The simultaneous substantial, but unequal by gender, improvement in mortality resulted in a reduced male/female mortality ratio, from 1.67 in the general population to 1.40. The reduced male/female mortality ratio was especially prominent for cancer and was not evident for heart disease mortality. Survival was nominally higher in non-White than in White participants. In summary, high-level employment is associated with substantially reduced mortality in both men and women. The relative improvement in survival is greater in men despite a comparable reduction in risk of heart disease mortality by gender. PMID- 11479187 TI - Effects of an angry temperament on coronary heart disease risk : The Atherosclerosis Risk in Communities Study. AB - The objective of the study was to determine which component of an anger-prone personality more strongly predicts coronary heart disease (CHD) risk. Proneness to anger, as assessed by the Spielberger Trait Anger Scale, is composed of two distinct subcomponents-anger-temperament and anger-reaction. Participants were 12,990 middle-aged Black men and women and White men and women from the Atherosclerosis Risk in Communities Study who were followed for the occurrence of acute myocardial infarction (MI)/fatal CHD, silent MI, or cardiac revascularization procedures (average = 53 months; maximum = 72 months) through December 31, 1995. Among normotensive persons, a strong, angry temperament (tendency toward quick, minimally provoked, or unprovoked anger) was associated with combined CHD (acute MI/fatal CHD, silent MI, or cardiac revascularization procedures) (multivariate-adjusted hazard ratio = 2.10, 95% confidence interval: 1.34, 3.29) and with 'hard" events (acute MI/fatal CHD) (multivariate adjusted hazard ratio = 2.28, 95% confidence interval: 1.29, 4.02). CHD event-free survival among normotensives who had a strong, angry temperament was not significantly different from that of hypertensives at either level of anger. These data suggest that a strong, angry temperament rather than anger in reaction to criticism, frustration, or unfair treatment places normotensive, middle-aged persons at increased risk for cardiac events and may confer a CHD risk similar to that of hypertension. PMID- 11479188 TI - Prevalence of and risk factors for abdominal aortic aneurysms in a population based study : The Tromso Study. AB - In a population-based study of 6,386 men and women aged 25--84 years in Tromso, Norway, in 1994--1995, the authors assessed the age- and sex-specific distribution of the abdominal aortic diameter and the prevalence of and risk factors for abdominal aortic aneurysm. Renal and infrarenal aortic diameters were measured with ultrasound. The mean infrarenal aortic diameter increased with age. The increase was more pronounced in men than in women. The age-related increase in the median diameter was less than that in the mean diameter. An aneurysm was present in 263 (8.9%) men and 74 (2.2%) women (p < 0.001). The prevalence of abdominal aortic aneurysm increased with age. No person aged less than 48 years was found with an abdominal aortic aneurysm. Persons who had smoked for more than 40 years had an odds ratio of 8.0 for abdominal aortic aneurysm (95% confidence interval: 5.0, 12.6) compared with never smokers. Low serum high density lipoprotein cholesterol was associated with an increased risk for abdominal aortic aneurysm. Other factors associated with abdominal aortic aneurysm were a high level of plasma fibrinogen and a low blood platelet count. Antihypertensive medication (ever use) was significantly associated with abdominal aortic aneurysm, but high systolic blood pressure was a risk factor in women only. This study indicates that risk factors for atherosclerosis are also associated with increased risk for abdominal aortic aneurysm. PMID- 11479189 TI - Predictors of asthma in young children: does reporting source affect our conclusions? AB - Both the size and statistical significance of sociodemographic and early health risk factors on childhood asthma vary across studies, in part because some studies rely on parents' retrospective reports of health conditions while others are based on medical records. The authors compare predictors of asthma alternately using maternal reports and medical records for the same set of children. Data are from the 1988 National Maternal and Infant Health Survey and 1991 Longitudinal Follow-up, which collected information from birth certificates, medical records, and mothers of a nationally representative, population-based cohort, allowing comparison across data sources for a consistent sample of young children in the United States. Concordance between maternal reports and medical records on asthma is moderate (kappa = 0.48). The authors find considerable discrepancies in both the estimated prevalence of asthma and the distribution across children with different sociodemographic and health characteristics, depending on the source of asthma data. Black race, male gender, and preterm birth are found to be risk factors for asthma regardless of data source. Poverty, large family size, urban residence, maternal smoking, and breastfeeding are significantly associated with asthma based on maternal reports but not medical records. Lower health care utilization among poor, uninsured, and urban children may account for part of the discrepancy. PMID- 11479190 TI - Association among physical activity level, cardiorespiratory fitness, and risk of musculoskeletal injury. AB - To help public health practitioners promote physical activities with a low risk of injury, this study determined the relation among type and duration of physical activity, cardiorespiratory fitness, and musculoskeletal injury in a sample of adults enrolled in the Aerobics Center Longitudinal Study. Subjects included 4,034 men and 967 women who underwent a baseline physical examination between 1970 and 1985 and who returned a mailed follow-up survey in 1986. At baseline, a treadmill graded exercise test was used to measure cardiorespiratory fitness. At follow-up, subjects reported injuries and type and duration of physical activity in the preceding 12 months. Polytomous logistic regression was used to estimate the association among physical activity type and duration, cardiorespiratory fitness, and injury. The risk of sustaining an activity-related injury increased with higher duration of physical activity per week and cardiorespiratory fitness levels. Results suggest that cardiorespiratory fitness may be a surrogate for unmeasured components of physical activity, such as exercise intensity. Among walkers, increasing duration of activity per week was not associated with an increased risk of injury. Results suggest that, for most adults, walking is a safe form of physical activity associated with a lower risk of injury than running or sport participation. PMID- 11479191 TI - New estimator of the genotype risk ratio for use in case-parental control studies. AB - Estimation of the genotype risk ratio can be an important part of studying the role of genetics in disease causation. For example, one might estimate risk among persons with genotype DD compared with risk among those with genotype Dd, where the candidate locus has alleles D and d, with D representing the disease susceptibility allele. In this paper, the authors propose a modified method of analysis for case-parental control studies that can improve efficiency. They show how investigators can use information from families in which both parents are observed to improve the estimator created by Sun et al., which applies when only one parent and an affected offspring have been observed. Since this information is not used by the conditional approach of Schaid and Sommer, the authors' approach allows for more complete use of available information, leading to a smaller mean squared error of the genotype risk ratio estimators. The authors also suggest a way to combine estimates from families in which one parent and one offspring are observed and estimates from families in which both parents and one offspring are observed. PMID- 11479192 TI - Application of nonparametric models for calculating odds ratios and their confidence intervals for continuous exposures. AB - Calculating odds ratios and corresponding confidence intervals for exposures that have been measured using a continuous scale presents important limitations in the traditional practice of analytical epidemiology. Approximations based on linear models require making arbitrary assumptions about the shape of the relation curve or about its breakpoints. Categorical analyses generally have low statistical efficiency, and cutpoints for the categories are in most cases arbitrary and/or opportunistic. The use of logistic generalized additive models to calculate odds ratios does not require these assumptions and allows great flexibility and adequate statistical efficiency. Based on the asymptotic normality of the logarithm of the odds ratio, the authors propose the use of an approximate analytical expression for the corresponding covariance matrix, which will allow the construction of confidence intervals for odds ratios that can be interpreted as in the classical parametric context. The authors illustrate this procedure by examining the relation between glycemia and risk of postoperative infection, using data obtained from a cohort study of patients undergoing surgery in Santiago, Spain (January 1996--March 1997). The authors found that glycemia values below 75 mg/dl and above 130 mg/dl were associated with increased risk of postoperative infection. PMID- 11479193 TI - Identifying confounding by indication through blinded prospective review. AB - Confounding by indication is a relentless threat to validity in observational studies of treatment effects. Multivariable models allow adjustment for known and readily measurable prognostic factors, but they may incompletely or inaccurately represent the underlying overall perceived risk of treatment. To incorporate practitioners' judgments about treatment indication and preprocedural prognosis into an observational study of cerebral aneurysm treatments, the author and colleagues presented patient characteristics and radiographic images from 179 aneurysm cases (University of California, San Francisco, 1990--1997) to panels of practitioners who were blinded as to actual treatment selection and outcome. In this way, the review process was designed to recreate the presentation of information in a prospective study. Judgments about inclusion and prognosis were reproducible. Perceived prognosis correlated with complication rates and provided information not present in a multivariable model including all available clinical characteristics. The association between treatment modality and outcome was examined while stratifying and adjusting for differences in perceived prognosis. Blinded prospective review may provide an unbiased observational study design with which to define a cohort that could have received any of the treatments being compared and to measure and adjust for overall perceived procedural risk. PMID- 11479194 TI - Re: "Biased tests of association: comparisons of allele frequencies when departing from Hardy-Weinberg proportions". PMID- 11479195 TI - Re: aluminum in drinking water and cognitive decline in elderly subjects: the Paquid cohort. PMID- 11479196 TI - Re: "Lung cancer among industrial sand workers exposed to crystalline silica". PMID- 11479197 TI - Selective estrogen receptor modulation: a personal perspective. PMID- 11479198 TI - PTEN-independent induction of caspase-mediated cell death and reduced invasion by the focal adhesion targeting domain (FAT) in human astrocytic brain tumors which highly express focal adhesion kinase (FAK). AB - Increased expression of focal adhesion kinase (FAK) was consistently observed in low- and high-grade astrocytomas and during glioblastoma progression after radiotherapy, but not in the more benign oligodendroglioma. In glioblastoma cell lines deficient for p53, p16(INK4A), and p14(ARF), FAK was inhibited in a dominant-negative manner by the focal adhesion targeting (FAT) domain, reducing invasion. In addition, caspase-3 activity was increased after serum withdrawal, or by cisplatin in the presence of serum, or upon loss of substrate attachment, and was in each case independent of PTEN status. Our results identify FAK as a potential target for anti-invasive strategies against infiltrating glioma cells. PMID- 11479199 TI - Expression profiling reveals hepsin overexpression in prostate cancer. AB - Prostate cancer is the most commonly diagnosed noncutaneous cancer in men. Despite this fact, many of the genetic changes that coincide with prostate cancer progression remain enigmatic. We have addressed this problem by characterizing the expression profiles of several benign and malignant human prostate samples, and we have identified several genes that are differentially expressed between benign and malignant glands. One gene that was overexpressed encodes the serine protease hepsin. We used an independent sample set to confirm that hepsin is overexpressed in prostate tumors, and in situ hybridization demonstrates that hepsin is specifically overexpressed in the carcinoma cells themselves. These facts, together with the molecular properties of hepsin, make it an ideal target for prostate cancer therapy. PMID- 11479200 TI - A SAGE (serial analysis of gene expression) view of breast tumor progression. AB - To identify molecular alterations involved in the initiation and progression of breast carcinomas, we analyzed the global gene expression profiles of normal mammary epithelial cells and in situ, invasive, and metastatic breast carcinomas using serial analysis of gene expression (SAGE). We identified sets of genes expressed only or most abundantly in a specific stage of breast tumorigenesis or in a certain subtype of tumors through the pair-wise comparison and by hierarchical clustering analysis of these eight SAGE libraries (two/stage). On the basis of these comparisons, we made the following observations: Normal mammary epithelial cells showed the most distinct and least variable gene expression profiles. Many of the genes highly expressed in normal mammary epithelium and lost in carcinomas encoded secreted proteins, cytokines, and chemokines, implicating abnormal paracrine and autocrine signaling in the initiation of breast tumorigenesis. Very few genes were universally up-regulated in all tumors regardless of their stage and histological grade, indicating a high degree of diversity at the molecular level that likely reflects the clinical heterogeneity characteristic of breast carcinomas. Tumors of different histology type and stage had very distinct gene expression patterns. No genes seemed to be specific for metastatic or for in situ carcinomas. We found that the most dramatic and consistent phenotypic change occurred at the normal-to-in situ carcinoma transition. This observation, combined with the fact that many of the genes involved encode secreted, cell-nonautonomous factors, implies that the normal epithelium-to-in situ carcinoma transition may be the most promising target for cancer prevention and treatment. PMID- 11479201 TI - Overexpression of Id-1 protein is a marker for unfavorable prognosis in early stage cervical cancer. AB - Inhibitor of differentiation/DNA binding (Id) proteins are transcription factors, involved in cell cycle regulation and neoangiogenesis. Using immunohistochemistry, we investigated the prognostic influence of Id-1, Id-2, and Id-3 expression in 89 patients with cervical cancer stage pT(1b). In univariate and multivariate analysis, patients with strong or moderate expression of Id-1 had a significant shorter overall survival time (P = 0.0144, log-rank test) and disease-free survival time (P = 0.0107, log-rank test) compared with those with low or absent Id-1 expression. Id-1 expression is an independent prognostic marker in early-stage cervical cancer. PMID- 11479202 TI - Dioxin suppresses the checkpoint protein, MAD2, by an aryl hydrocarbon receptor independent pathway. AB - The compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown recently to be carcinogenic, but little is currently known about the molecular mechanism of TCDD affecting cell proliferation and carcinogenesis. In this report, we demonstrate that TCDD suppresses the expression of the checkpoint protein, Mad2. Suppression of Mad2 was also observed in aryl hydrocarbon receptor-deficient mouse embryonic fibroblasts, suggesting that TCDD suppresses Mad2 by a novel TCDD receptor signaling mechanism. In addition, HeLa cells treated with TCDD failed to arrest in mitosis after nocodazole treatment. The Mad2 protein plays a significant role in accurate chromosome segregation in mitotic cells. Our data suggest that TCDD may increase chromosomal instability through the suppression of Mad2 expression. PMID- 11479203 TI - Modest increased sensitivity to radiation oncogenesis in ATM heterozygous versus wild-type mammalian cells. AB - Subpopulations that are genetically predisposed to radiation-induced cancer could have significant public health consequences. Individuals homozygous for null mutations at the ataxia telangiectasia gene are indeed highly radiosensitive, but their numbers are very small. Ataxia Telangiectasia heterozygotes (1-2% of the population) have been associated with somewhat increased radiosensitivity for some end points, but none directly related to carcinogenesis. Here, intralitter comparisons between wild-type mouse embryo fibroblasts and mouse embryo fibroblasts carrying ataxia telangiectasia mutated (ATM) null mutation indicate that the heterozygous cells are more sensitive to radiation oncogenesis than their normal, litter-matched, counterparts. From these data we suggest that Ataxia Telangiectasia heterozygotes could indeed represent a societally significant radiosensitive human subpopulation. PMID- 11479204 TI - Hepatoma-derived growth factor is associated with reduced sensitivity to irradiation in esophageal cancer. AB - Radiotherapy is a useful component of treatment for esophageal cancer. Identification of the genes that are differentially expressed between radiosensitive and radioresistant cancer cells is important for predicting clinical effectiveness of radiotherapy. We established human esophageal cancer cell lines resistant to X-ray. Using differential display, we obtained one gene that was expressed in radiosensitive cells but was rarely expressed in radioresistant cells, and that gene was identical with hepatoma-derived growth factor (HDGF), an acidic polypeptide with mitogenic activity for fibroblasts. The semiquantitative reverse transcription-PCR assay confirmed that HDGF mRNA expression was reduced in established radioresistant cells, and its reduction was associated with reduced sensitivity to irradiation. Radiotherapy was more effective in clinical cases with high HDGF mRNA expression compared with cases with low expression (P < 0.05). The findings demonstrate that HDGF may play an important role in radiosensitivity, and it could be a novel marker predicting effectiveness of radiotherapy in clinical cases. PMID- 11479205 TI - p53, CHK2, and CHK1 genes in Finnish families with Li-Fraumeni syndrome: further evidence of CHK2 in inherited cancer predisposition. AB - Germ-line mutations in the p53 gene predispose individuals to Li-Fraumeni syndrome (LFS). The cell cycle checkpoint kinases CHK1 and CHK2 act upstream of p53 in DNA damage responses, and recently rare germ-line mutations in CHK2 were reported in LFS families. We have analyzed CHK1, CHK2, and p53 genes for mutations in 44 Finnish families with LFS, Li-Fraumeni-like syndrome, or families phenotypically suggestive of LFS with conformation-sensitive gel electrophoresis. Five different disease-causing mutations were observed in 7 families (7 of 44 families; 15.9%): 4 in the p53 gene (5 of 44 families; 11.4%) and 1 in the CHK2 gene (2 of 44 families; 4.5%). Interestingly, the other CHK2-mutation carrier also has a mutation in the MSH6 gene. The cancer phenotype in the CHK2-families was not characteristic of LFS, and may indicate variable phenotypic expression in the rare families with CHK2 mutations. No mutations in the CHK1 gene were identified. Additional work is necessary to completely unravel the molecular background of LFS. PMID- 11479206 TI - Prognostic significance of circulating microsatellite markers in the plasma of melanoma patients. AB - PURPOSE: Multiple genetic alterations including loss of heterozygosity (LOH) occur commonly in melanoma tumors. We demonstrated previously free-circulating DNA microsatellites with LOH in the blood of melanoma patients. These LOH markers in plasma may be useful as surrogates for subclinical disease progression. The purpose of this study was to determine whether the presence of circulating tumor microsatellite markers in the preoperative blood from patients with melanoma has prognostic utility. EXPERIMENTAL DESIGN: Plasma was analyzed for the presence of LOH at six chromosome regions, which are common for allelic loss in melanoma tumors, in 57 patients undergoing surgical resection of all of the clinically apparent disease. RESULTS: LOH was detected in 32 of 57 patients (56%). Both LOH incidence and frequency correlated with advancing American Joint Committee on Cancer stage. In patients with American Joint Committee on Cancer stage III, the presence of LOH as an independent variable in preoperative plasma was significantly associated (P = 0.05) with an increased risk of death. Furthermore, LOH at microsatellite marker D1S228 in the plasma of patients with advanced disease correlated significantly (P = 0.0009) with a poorer survival after surgical resection. LOH commonly found in melanoma tumors can be successfully identified in the plasma of a patient, providing a potentially less invasive route for following genetic changes that serve as molecular surrogates for assessing subclinical disease progression. CONCLUSIONS: This study provides evidence that blood testing for circulating tumor genetic markers may provide valuable prognostic information and guide future therapy. PMID- 11479207 TI - Aberrant methylation and simian virus 40 tag sequences in malignant mesothelioma. AB - Aberrant promoter methylation and resultant silencing of several genes plays an important role in the pathogenesis of many tumor types. We compared the methylation profile of 66 malignant mesotheliomas (MMs) and 40 lung adenocarcinomas using methylation-specific PCR for seven genes frequently methylated in lung cancer. We also compared the methylation frequencies of these genes as well as the methylation index, a reflection of all of the gene frequencies, with the presence of SV40 large T-antigen (Tag) sequences, histological subtype, and patient survival. Our major findings are: (a) with the exception of the RASSF1A promoter of the RASSF1 gene, frequencies of aberrant methylation were significantly lower in MMs than in adenocarcinomas; (b) the frequency of RASSF1A aberrant methylation and the value of the methylation index were significantly higher in SV40 sequence positive MM than in negative MM; and (c) the methylation index was higher in epithelial MM than in sarcomatous/mixed MM. Our results demonstrate a relationship between SV40 and aberrant methylation in MMs. PMID- 11479208 TI - HIF-1alpha-mediated up-regulation of vascular endothelial growth factor, independent of basic fibroblast growth factor, is important in the switch to the angiogenic phenotype during early tumorigenesis. AB - The switch to the angiogenic phenotype represents a critical checkpoint during tumor progression. The acquisition of new capillary vessels provides newly vascularized tumor nodules with a distinct biological advantage over their avascular counterparts by conferring upon them the ability to expand and develop both locally and metastatically. To identify the molecules and mechanisms underlying this rate-limiting step in successful tumorigenesis, we have developed an in vivo tumor model that reproducibly recapitulates the angiogenic switch. Using this model, we have analyzed vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and hypoxia-inducible factor-1alpha (HIF 1alpha) expression and activity in both avascular and vascular growth phases of the tumor. A significantly higher level of VEGF protein was detected in avascular tumor nodules compared with vascular nodules. As avascular tumors became vascularized, VEGF levels decreased approximately 10-fold. In contrast, bFGF levels were not elevated in avascular nodules but rather were detected at levels approximately 2 times higher in vascular nodules compared with the avascular tumor nodules. Given that VEGF is transcriptionally regulated by HIF-1alpha, immunohistochemical studies of chondrosarcoma nodules were conducted and revealed that the nuclear translocation of HIF-1alpha was detected exclusively in avascular tumor nodules. This study implicates HIF-1alpha-mediated up-regulation of VEGF but not bFGF in the switch to the angiogenic phenotype during tumorigenesis. PMID- 11479209 TI - Vascular endothelial growth factor is an autocrine survival factor for neuropilin expressing breast carcinoma cells. AB - We identify a novel function for the vascular endothelial growth factor (VEGF) in its ability to stimulate an autocrine signaling pathway in metastatic breast carcinoma cells that is essential for their survival. Suppression of VEGF expression in metastatic cells in vitro induced their apoptosis, in addition to inhibiting the constitutively elevated phosphatidylinositol 3'-kinase activity that is characteristic of these cells and important for their survival. Hypoxia enhanced the survival of metastatic cells by increasing VEGF expression. The importance of the VEGF receptor neuropilin was indicated by the ability of a neuropilin-binding VEGF isoform to enhance breast carcinoma survival. Moreover, the expression of neuropilin in neuropilin-deficient breast carcinoma cells protected them from apoptosis. The identification of this VEGF autocrine signaling pathway has important implications for tumor metastasis and therapeutic intervention. PMID- 11479210 TI - A monoclonal antibody that induces neuronal apoptosis binds a metastasis marker. AB - The cell surface molecules controlling apoptosis in cortical neurons are largely unknown. A monoclonal antibody was derived that induces cultured neocortical neurons to undergo apoptosis. A Fab fragment of the antibody, however, lacked the ability to induce cell death. The antigen was purified, and characterized by compositional analysis, fast atom bombardment (FAB) mass spectrometry, sequential exoglycosidase treatments, methylation analysis, and (1)H-nuclear magnetic resonance spectroscopy, proving to be isoglobotetraosylceramide (IsoGb4). IsoGb4 has been shown previously to be a metastasis marker, antibodies against which block metastases in a mammary adenocarcinoma model (S. A. Carlsen et al., Cancer Res., 53: 2906-2911, 1993). Addition of the purified antigen to cells lacking this glycolipid demonstrated that it is capable of functioning as a portable apoptosis-transducing molecule. Intracellular ceramide levels were increased after the treatment with the apoptosis-inducing antibody, but the membrane sphingomyelin level remained unchanged. Fumonisin B1 inhibited both the ceramide increase and the apoptosis induced via IsoGb4, which indicated that the ceramide synthase pathway is likely to be involved in apoptosis induction by IsoGb4. PMID- 11479211 TI - Two novel glycosides from the fruits of Morinda citrifolia (noni) inhibit AP-1 transactivation and cell transformation in the mouse epidermal JB6 cell line. AB - The fruit juice of Morinda citrifolia (noni), a plant originally grown in the Hawaiian and Tahitian islands, has long been used by islanders to treat diseases, including cancer. Two novel glycosides, 6-O-(beta-D-glucopyranosyl)-1-O-octanoyl beta-D-glucopyranose and asperulosidic acid, extracted from the juice of noni fruits, were used to examine their effects on 12-O-tedtradecanoylphorbol-13 acetate (TPA)- and epidermal growth factor (EGF)-induced AP-1 transactivation and cell transformation in mouse epidermal JB6 cells. The results indicated that both compounds were effective in suppressing TPA- or EGF-induced cell transformation and associated AP-1 activity. TPA- or EGF-induced phosphorylation of c-Jun, but not extracellular signal-regulated kinases or p38 kinases, was also blocked by the compounds, indicating that c-Jun N-terminal kinases were critical in mediating TPA- or EGF-induced AP-1 activity and subsequent cell transformation in JB6 cells. PMID- 11479212 TI - Interactions between methylating and pyridyloxobutylating agents in A/J mouse lungs: implications for 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis. AB - The tobacco-specific nitrosamine, 4-(methylnitrosoamino)-1-(3-pyridyl)-1 butanone, is activated to lung DNA methylating and pyridyloxobutylating intermediates. It is likely that both pathways play a role in lung tumor initiation by this nitrosamine. Previous studies indicated that O(6) methylguanine (O(6)-mG) persistence is critical for lung tumor formation in A/J mice. The model pyridyloxobutylating agent, 4-(acetoxymethylnitrosamino)-1-(3 pyridyl)-1-butanone (NNKOAc), enhanced the tumorigenic activity of a model methylating agent, acetoxymethylmethylnitrosamine (AMMN), presumably by increasing O(6)-mG persistence in lung DNA. We have been testing the hypothesis that the pyridyloxobutylation pathway increases the mutagenic activity of the DNA methylation pathway by preventing the repair of O(6)-mG by O(6)-alkylguanine-DNA alkyltransferase (AGT). In this study, we report that NNKOAc depletes AGT in lungs but not livers of A/J mice. The consequences of AGT depletion by NNKOAc were then compared with those observed with a known AGT inhibitor, O(6) benzylguanine (O(6)-bG). NNKOAc and O(6)-bG had similar effects on the levels of AMMN-derived O(6)-mG at 4 and 96 h postinjection. This increase in O(6)-mG levels correlated to increased lung tumor multiplicity in animals simultaneously treated with AMMN (0.75 or 1 micromol) and NNKOAc or O(6)-bG. Only NNKOAc significantly increased lung tumor multiplicity at doses of 0.25 or 0.5 micromol AMMN. The results from these studies indicate that the pyridyloxobutylating agent, NNKOAc, can influence the tumorigenic activity of methylating agents in two ways. At low AMMN doses, the increase in tumor multiplicity is dominated by the additive tumorigenic properties of AMMN and NNKOAc. At higher AMMN doses, NNKOAc appears to enhance the tumorigenic activity of AMMN through enhanced depletion of the repair protein, AGT, leading to increased O(6)-mG persistence. It is likely that similar interactions are important for the organospecific effects of 4 (methylnitrosoamino)-1-(3-pyridyl)-1-butanone. PMID- 11479213 TI - Preferential growth stimulation of mammary glands over uterine endometrium in female rats by a naturally occurring estradiol-17beta-fatty acid ester. AB - We hypothesize that the endogenously present lipoidal estrogen fatty acid esters may have a stronger mitogenic action in the fat-rich mammary tissues than in the uterus. To test this hypothesis, we compared the activity of estradiol-17beta stearate (E(2)-17beta-S) with that of estradiol-17beta (E(2)) in stimulating the growth of mammary glandular cells versus the growth of uterine endometrial cells in ovariectomized female Sprague Dawley rats. Experimentally, an estimated 0.5 or 5 nmol of E(2)-17beta-S or E(2) was released daily to ovariectomized female rats through an Alzet pump implanted under the back skin of the animal for 10 or 23 days. The growth-stimulatory effect of E(2)-17beta-S and E(2) on mammary glandular cells was determined according to 5-bromo-2'-deoxyuridine labeling indices, and their effect on the uterus was determined by measuring both the 5 bromo-2'-deoxyuridine labeling index and the uterine wet weight. Our results showed that chronic treatment of ovariectomized female rats with 0.5 or 5 nmol/day E(2)-17beta-S for 10 or 23 days had a stronger stimulatory effect on mammary glandular cell proliferation than treatment with equimolar doses of E(2). In the uterus, however, E(2) was more active in stimulating the proliferation of uterine endometrial cells than E(2)-17beta-S at equimolar doses. Our results demonstrated, for the first time, that a naturally occurring estradiol-17beta fatty acid ester has a differential, strong mitogenic effect in the fat-rich mammary tissues, and this effect was not observed with E(2). It is tempting to suggest that the fatty acid esters of the endogenous estrogens and their bioactive metabolites (e.g., 4-hydroxyestradiol and 16alpha-hydroxyestrone) may be of unique importance for stimulating cell growth and possibly also for inducing tumor formation in the fat-rich mammary tissues as compared with the uterus. More studies are warranted to test these ideas. PMID- 11479214 TI - Re-expression of estrogen receptor alpha in estrogen receptor alpha-negative MCF 7 cells restores both estrogen and insulin-like growth factor-mediated signaling and growth. AB - Estrogen can increase insulin-like growth factor-I receptor (IGF-IR) and insulin receptor substrate-1 (IRS-1) expression, two key components of IGF-I-mediated signaling. The result is sensitization of breast cancer cells to IGF-I and synergistic growth in the presence of estrogen and IGF-I. We hypothesized that loss of estrogen receptor alpha (ERalpha) would result in reduced IGF-mediated signaling and growth. To test this hypothesis, we examined IGF-I effects in MCF-7 breast cancer cell sublines that have been selected for loss of ERalpha (C4 and C4-12 cells are ERalpha-negative) by long-term estrogen withdrawal. C4 and C4-12 cells had reduced IGF-IR and IRS-1 mRNA and protein expression (compared with MCF 7 cells) that was not inducible by estrogen. Furthermore, C4 and C4-12 cells showed reduced IGF-I signaling and failed to show any growth response to either estrogen or IGF-I. To prove that loss of IGF and estrogen-mediated signaling and growth was a consequence of loss of ERalpha, we re-expressed ERalpha in C4-12 cells by stable transfection with HA-tagged ERalpha. Three independent C4-12 ERalpha-HA clones expressed a functional ERalpha that (a) was down-regulated by estrogen, (b) conferred estrogen-induction of cyclin D1 expression, and (c) caused estrogen-mediated increase in the number of cells in S phase. All of the effects were completely blocked by antiestrogens. Interestingly, ERalpha-HA expression in C4-12 cells did not restore estrogen induction of progesterone receptor expression. However, ERalpha-positive C4-12 cells now exhibited estrogen induction of IGF-IR and IRS-1 levels and responded mitogenically to both estrogen and IGF-I. These data show that ERalpha is a critical requirement for IGF signaling, and to our knowledge this is the first report of functional ERalpha expression that confers estrogen-mediated growth of an ER-negative breast cancer cell line. PMID- 11479215 TI - Growth inhibition of dermatofibrosarcoma protuberans tumors by the platelet derived growth factor receptor antagonist STI571 through induction of apoptosis. AB - Dermatofibrosarcoma protuberans (DFSP) and giant cell fibroblastoma (GCF) are recurrent, infiltrative skin tumors that presently are treated with surgery. DFSP and GCF tumors are genetically characterized by chromosomal rearrangements fusing the collagen type Ialpha1 (COLIA1) gene to the platelet-derived growth factor B chain (PDGFB) gene. It has been shown that the resulting COL1A1/PDGF-B fusion protein is processed to mature PDGF-BB. Autocrine PDGF receptor stimulation has therefore been predicted to contribute to DFSP and GCF tumor development and growth. Here we demonstrate presence of activated PDGF receptors in primary cultures derived from six different DFSP and GCF tumors. Three of the primary cultures were further characterized; their in vitro growth displayed an increased sensitivity to treatment with the PDGF receptor tyrosine kinase inhibitor STI571, as compared with normal fibroblasts. Transplantable tumors, displaying a DFSP like histology, were established from one of the DFSP primary cultures. Treatment of tumor-bearing severe combined immunodeficient mice with STI571 reduced tumor growth. The growth-inhibitory effects in vitro and in vivo occurred predominantly through induction of tumor cell apoptosis. Our study demonstrates growth inhibitory effects of PDGF receptor antagonists on human DFSP- and GCF-derived tumor cells and demonstrates that autocrine PDGF receptor stimulation provides antiapoptotic signals contributing to the growth of these cells. These findings suggest targeting of PDGF receptors as a novel treatment strategy for DFSP and GCF. PMID- 11479216 TI - Inhibition of glioma growth in vivo by selective activation of the CB(2) cannabinoid receptor. AB - The development of new therapeutic strategies is essential for the management of gliomas, one of the most malignant forms of cancer. We have shown previously that the growth of the rat glioma C6 cell line is inhibited by psychoactive cannabinoids (I. Galve-Roperh et al., Nat. Med., 6: 313-319, 2000). These compounds act on the brain and some other organs through the widely expressed CB(1) receptor. By contrast, the other cannabinoid receptor subtype, the CB(2) receptor, shows a much more restricted distribution and is absent from normal brain. Here we show that local administration of the selective CB(2) agonist JWH 133 at 50 microg/day to Rag-2(-/-) mice induced a considerable regression of malignant tumors generated by inoculation of C6 glioma cells. The selective involvement of the CB(2) receptor in this action was evidenced by: (a) the prevention by the CB(2) antagonist SR144528 but not the CB(1) antagonist SR141716; (b) the down-regulation of the CB(2) receptor but not the CB(1) receptor in the tumors; and (c) the absence of typical CB(1)-mediated psychotropic side effects. Cannabinoid receptor expression was subsequently examined in biopsies from human astrocytomas. A full 70% (26 of 37) of the human astrocytomas analyzed expressed significant levels of cannabinoid receptors. Of interest, the extent of CB(2) receptor expression was directly related with tumor malignancy. In addition, the growth of grade IV human astrocytoma cells in Rag-2( /-) mice was completely blocked by JWH-133 administration at 50 microg/day. Experiments carried out with C6 glioma cells in culture evidenced the internalization of the CB(2) but not the CB(1) receptor upon JWH-133 challenge and showed that selective activation of the CB(2) receptor signaled apoptosis via enhanced ceramide synthesis de novo. These results support a therapeutic approach for the treatment of malignant gliomas devoid of psychotropic side effects. PMID- 11479217 TI - Signaling-inactive epidermal growth factor receptor/ligand complexes in intact carcinoma cells by quinazoline tyrosine kinase inhibitors. AB - Several inhibitors of EGF receptor (EGFR) tyrosine kinase activity have been developed that compete with ATP at its binding site such as the quinazolines PD 153035 and ZD 1839 or the 4,5-dianilino-phthalimides DAPH1 and DAPH2. When tested on human A431 cells, the quinazolines completely blocked EGF-induced receptor phosphorylation at 100 nM, whereas it was inhibited by DAPH1 and DAPH2 by only 20% at 3 microM. Quinazoline-treated A431 as well as tumor cells expressing less EGFR (A549, MDA MB 231, and T47D) bound 3- to 6-fold more (125)I-labeled EGF than untreated intact control cells. Scatchard analysis revealed the disappearance of low- and high-affinity EGFR on A431 cells upon PD 153035 treatment. A single receptor class of intermediate ligand binding affinity emerged and its number corresponded to the sum of the two classes. DAPH1 and DAPH2 did not change ligand binding properties of EGFR. PD 153035 exerted the most potent effects on EGF binding to A431 or on inhibiting EGF-stimulated growth of rat MTLn3 cells at low ligand concentrations. Cross-linking of EGFR on PD 153035-treated A431 cells indicated the formation of inactive dimers that further increased upon addition of EGF. Chemical cross-linking of (125)I-labeled EGF to PD 153035-treated A431 cells revealed increased binding to monomeric and dimeric EGFR. Thus, the quinazolines sequestered EGFR plus the ligand into inactive receptor/ligand complexes. This novel mode of action of quinazoline tyrosine kinase inhibitors may be the basis for their extraordinary potency especially in conditions when the ligand is present in limiting amounts. PMID- 11479218 TI - Treatment of malignant glioma cells with the transfer of constitutively active caspase-6 using the human telomerase catalytic subunit (human telomerase reverse transcriptase) gene promoter. AB - Because the apoptotic pathway is often disrupted in tumor cells, its genetic restoration is a very attractive approach for the treatment of tumors. To treat malignant gliomas with this approach, it would be preferred to restrict induction of apoptosis to tumor cells by establishing a tumor-specific expression system. Telomerase is an attractive target because the vast majority of malignant gliomas have telomerase activity whereas normal brain cells do not. Activation of telomerase is tightly regulated at the transcriptional level of the telomerase catalytic subunit [human telomerase reverse transcriptase, (hTERT)]. Therefore, we hypothesized that using a hTERT promoter-driven vector system, an apoptosis inducible gene may be preferentially restricted to telomerase- or hTERT-positive tumor cells. In this study, we constructed an expression vector consisting of the constitutively active caspase-6 (rev-caspase-6) under the hTERT promoter (hTERT/rev-caspase-6) and then investigated its antitumor effect on malignant glioma cells. The rationale for using the rev-caspase-6 gene is because it induces apoptosis independent of the initiator caspases. We demonstrated that the hTERT/rev-caspase-6 construct induced apoptosis in hTERT-positive malignant glioma cells, but not in hTERT-negative astrocytes, fibroblasts, and alternative lengthening of telomeres cells. In addition, the growth of s.c. tumors in nude mice was significantly suppressed by the treatment with hTERT/rev-caspase-6 construct. The present results strongly suggest that the telomerase-specific transfer of the rev-caspase-6 gene under the hTERT promoter is a novel targeting approach for the treatment of malignant gliomas. PMID- 11479219 TI - Multiple microtubule alterations are associated with Vinca alkaloid resistance in human leukemia cells. AB - Vinca alkaloids are used extensively in the treatment of childhood acute lymphoblastic leukemia (ALL) and despite their usefulness, drug resistance remains a serious clinical problem. Vinca alkaloids bind to the beta-tubulin subunit of the alpha/beta-tubulin heterodimer and inhibit polymerization of microtubules. Recent studies have implicated altered beta-tubulin isotype expression and mutations in resistance to microtubule-stabilizing agents. Microtubule-associated protein (MAP) MAP4 binds to and stabilizes microtubules, and increased expression is associated with decreased sensitivity to microtubule depolymerizing agents. To address the significance of beta-tubulin and MAP4 alterations in childhood ALL, two CCRF-CEM-derived Vinca alkaloid resistant cell lines, VCR R (vincristine) and VLB100 (vinblastine), were examined. Decreased expression of class III beta-tubulin was detected in both VCR R and VLB100 cells. VCR R cells and to a lesser extent VLB100 cells expressed increased levels of MAP4 protein. Increased microtubule stability was observed in these VCR R cells as identified by the high levels of polymerized tubulin (45.6 +/- 2.6%; P < 0.005) compared with CEM and VLB100 cells (24.7 +/- 3.3% and 24.7 +/- 2.5%, respectively). Expression was associated with a single MAP4 isoform in the polymerized microtubule fraction in CEM and VCR cells. In contrast, VLB100 cells expressed a lower molecular weight isoform in the polymerized fraction. Two dimensional-PAGE and immunoblotting revealed marked posttranslational changes in class I beta-tubulin in VCR R cells not evident in CEM cells. Sequencing of the beta-tubulin (HM40) gene identified a point mutation in VCR R cells in nucleotide 843 (CTC-->ATC; Leu(240)-->Ile) that was not present in CEM or VLB100 cells. This mutation resides in a region of beta-tubulin that lies in close proximity to the alpha/beta tubulin interface. Multiple alterations related to normal microtubule function were identified in ALL cells selected for resistance to Vinca alkaloids, and these alterations may provide important insight into mechanisms mediating resistance to Vinca alkaloids. PMID- 11479220 TI - Differing contribution of thiopurine methyltransferase to mercaptopurine versus thioguanine effects in human leukemic cells. AB - Thioguanine and mercaptopurine are prodrugs requiring conversion into thiopurine nucleotides to exert cytotoxicity. Thiopurine S-methyltransferase (TPMT), an enzyme subject to genetic polymorphism, catabolizes thiopurines into inactive methylated bases, but also produces methylthioguanine nucleotides and methylmercaptopurine nucleotides from thioguanine and mercaptopurine nucleotides, respectively. To study the effect of TPMT on activation versus inactivation of mercaptopurine and thioguanine, we used a retroviral gene transfer technique to develop human CCRF-CEM cell lines that did (TPMT+) and did not (MOCK) overexpress TPMT. After transduction, TPMT activities were 14-fold higher in the TPMT+ versus the MOCK cell lines (P < 0.001). TPMT+ cells were less sensitive to thioguanine than MOCK cells (IC(50) = 1.10+/- 0.12 microM versus 0.55 +/- 0.19 microM; P = 0.02); in contrast, TPMT+ cells were more sensitive to mercaptopurine than MOCK cells (IC(50) = 0.52 +/- 0.20 microM versus 1.50 +/- 0.23 microM; P < 0.01). The lower sensitivity of TPMT+ versus MOCK cells to thioguanine was associated with lower thioguanine nucleotide concentrations (917 +/- 282 versus 1515 +/- 183 pmol/5 x 10(6) cells; P = 0.01), higher methylthioguanine nucleotide concentrations (252 +/- 34 versus 27 +/- 10 pmol/5 x 10(6) cells; P = 0.01), less inhibition of de novo purine synthesis (13 versus 95%; P < 0.01), and lower deoxythioguanosine incorporation into DNA (2.0 +/- 0.6% versus 7.2 +/- 2.0%; P < 0.001). The higher sensitivity of TPMT+ cells to mercaptopurine was associated with higher concentrations of methylmercaptopurine nucleotide (2601 +/- 1055 versus 174 +/- 77 pmol/5 x 10(6) cells; P = 0.01) and greater inhibition of de novo purine synthesis (>99% versus 74%; P < 0.01) compared with MOCK cells. We conclude that methylation of mercaptopurine contributes to the antiproliferative properties of the drug, probably through inhibition of de novo purine synthesis by methylmercaptopurine nucleotides, whereas thioguanine is inactivated primarily by TPMT. PMID- 11479221 TI - Helenalin triggers a CD95 death receptor-independent apoptosis that is not affected by overexpression of Bcl-x(L) or Bcl-2. AB - Apoptosis is required for proper tissue homeostasis. Defects in apoptosis signaling pathways, thus, contribute to carcinogenesis and chemoresistance. A major goal in chemotherapy is, therefore, to find cytotoxic agents that restore the ability of tumor cells to undergo apoptosis. We show here that the sesquiterpene lactone helenalin (10-50 microM) induces apoptosis in leukemia Jurkat T cells even if they lack the CD95 death receptor or overexpress the antiapoptotic proteins Bcl-x(L) or Bcl-2. Activated peripheral blood mononuclear cells, however, are not affected (10-50 microM helenalin). Helenalin led to a time-dependent (0-24 h) cleavage of the specific caspase-3-like substrate Asp-Glu Val-Asp-7-amino-4-trifluoromethylcoumarin as well as to the proteolytic processing of procaspase-3 and -8. Caspase activation was a necessary requirement for apoptosis because the broad-spectrum caspase inhibitor benzyloxycarbonyl-Val Ala-Asp(OMe)-fluoromethylketone (zVAD-fmk, 50 microM) completely abrogated helenalin-induced DNA fragmentation as well as phosphatidylserin translocation. Although the initiator caspase-8 was activated, the helenalin-induced signaling pathway did not require the CD95 death receptor as shown using cells without or with an antibody (ZB4)-blocked CD95 receptor. Helenalin also did not induce CD95 or CD95-ligand expression. On the other hand, helenalin was found to induce the release of cytochrome c from mitochondria that was not inhibited by the caspase inhibitor zVAD-fmk, which indicated that cytochrome c release precedes caspase activation. Cytochrome c release was accompanied by dissipation of the mitochondrial transmembrane potential (DeltaPsi(m)), which was partly inhibited by zVAD-fmk, which suggests that caspases are involved in loss of DeltaPsi(m). Most importantly, overexpression of the mitochondria protecting proteins Bcl-x(L) or Bcl-2 failed to confer resistance to helenalin-induced apoptosis, although the data presented here suggest that helenalin induces a mitochondria-dependent pathway. Thus, helenalin is a promising experimental cytotoxic agent that possibly points to new strategies to overcome apoptosis resistance attributable to overexpression of antiapoptotic Bcl-2 proteins. PMID- 11479222 TI - Antitumor effect of 5-aminolevulinic acid-mediated photodynamic therapy can be enhanced by the use of a low dose of photofrin in human tumor xenografts. AB - Practically all of the exogenous photosensitizers used for clinical photodynamic therapy (PDT) target mainly vasculature. Although effective in tumor destruction, they also, unavoidably, induce phototoxicity of normal tissues. Porphyrins synthesized endogenously from 5-aminolevulinic acid (ALA) accumulate within cells. Tumor eradication would be more efficient if both cellular components and vascular stroma of a tumor could be targeted. Thus, PDT with a mixture of ALA and Photofrin (Pf, a vessel-targeted sensitizer) may simultaneously destroy the two elements. Using chemical extraction assays, pharmacokinetics of ALA and ALA induced porphyrins were studied in the plasma and tumors of nude mice bearing human WiDr and KM20L2 colonic carcinomas after an i.p. injection of 250 mg/kg body weight of ALA. Subsequently, PDT efficacy of the two tumor models with ALA, Pf, or with the two drugs in combination was evaluated. The phototoxic effects on tumor cells in vitro with the combined drugs was also determined. Moreover, histological and ultrastructural alterations of the treated tumors were investigated, and tumor cell clonogenicity was assessed as a function of time after in vivo PDT using an in vitro colony formation assay. Finally, the photosensitivity of normal skin tissue treated according to various protocols was compared. The amounts of ALA peaked at 0.5 h after administration in both plasma and WiDr tumor. The rates of ALA clearance seemed to follow a one-compartment model with half-lives of approximately 18 and 58 min in the plasma and tumor, respectively. About 100 and 60 times higher concentrations of ALA were needed to induce a given concentration of porphyrins in the plasma and tumor, respectively, although the plasma porphyrins may not only be released from blood cells but also from other organs. Similar kinetics of distribution patterns of ALA- and ALA methylester-induced porphyrins were found in the plasma and tumors, and the elimination rates were consistent with a two-compartment model. ALA induced much more porphyrins than ALA methylester in both plasma and tumors. Tumors PDT treated with ALA plus Pf at a low dose (1 mg/kg) grew significantly more slowly than those treated with either of the drugs in both WiDr and KM20L2 models. However, the enhanced antitumor effect was not found in the tumor cells under in vitro conditions. Morphological studies demonstrated that PDT with the combined regimen resulted in necrosis of neoplastic cells and severe disruption of tumor microvasculature. This was supported by the findings obtained from the studies of in vivo PDT and in vitro clonogenic assay that showed a progressive reduction in tumor cell viability with times following PDT. Such a combined PDT protocol did not induce any phototoxicity in normal skin tissue. These data indicate that targeting both neoplastic cells and stroma with ALA and Pf (a low dose) can potentiate antitumor PDT effect with no risk of prolonged skin photosensitivity. PMID- 11479223 TI - Chimeric tumor suppressor 1, a p53-derived chimeric tumor suppressor gene, kills p53 mutant and p53 wild-type glioma cells in synergy with irradiation and CD95 ligand. AB - Adenoviral chimeric tumor suppressor 1 (CTS1) gene transfer was evaluated as a novel approach of somatic gene therapy for malignant glioma. CTS1 is an artificial p53-based gene designed to resist various pathways of p53 inactivation. Here, we report that an adenovirus encoding CTS1 (Ad-CTS1) induces growth arrest and loss of viability in all glioma cell lines examined, in the absence of specific cell cycle changes. In contrast, an adenovirus encoding wild type p53 (Ad-p53) does not consistently induce apoptosis in the same cell lines. Electron microscopic analysis of Ad-CTS1-infected glioma cells reveals complex cytoplasmic pathology and delayed apoptotic changes. Ad-CTS1 induces prominent activation of various p53 target genes, including p21 and MDM-2, but has no relevant effects on BCL-2 family protein expression. Although Ad-CTS1 strongly enhances CD95 expression at the cell surface, endogenous CD95/CD95 ligand interactions do not mediate CTS1-induced cell death. This is because Ad-CTS1 promotes neither caspase activation nor mitochondrial cytochrome c release and because the caspase inhibitors, z-val-Ala-DL-Asp-fluoromethylketone (zVAD)-fmk or z-Ile-Glu-Thr-Asp- fluoromethylketone (z-IETD)-fmk, do not block CTS1-induced cell death. Ad-CTS1 synergizes with radiotherapy and CD95 ligand in killing glioma cells. In summary, Ad-CTS1 induces an unusual type of cell death that appears to be independent of BCL-2 family proteins, cytochrome c release, and caspases. CTS1 gene transfer is a promising strategy of somatic gene therapy for malignant glioma. PMID- 11479224 TI - Abrogation of the Chk1-mediated G(2) checkpoint pathway potentiates temozolomide induced toxicity in a p53-independent manner in human glioblastoma cells. AB - Temozolomide (TMZ) produces O(6)-methylguanine in DNA, which in turn mispairs with thymine, triggering futile DNA mismatch repair (MMR) and ultimately cell death. We found previously that in p53-proficient human glioma cells, TMZ-induced futile DNA MMR resulted not in apoptosis but rather in prolonged, p53- and p21 associated G(2)-M arrest and senescence. Additionally, p53-deficient cells were relatively more TMZ resistant than p53-deficient glioma cells, which underwent only transient G(2)-M arrest before death by mitotic catastrophe. These results suggested that prolonged G(2)-M arrest might protect cells from TMZ-induced cytotoxicity. In the present study, we therefore focused on the mechanism by which TMZ induces G(2)-M arrest and on whether inhibition of such G(2)-M arrest might sensitize glioma cells to TMZ-induced toxicity. U87MG glioma cells treated with TMZ underwent G(2)-M arrest associated with Chk1 activation and phosphorylation of both cdc25C and cdc2. These TMZ-induced effects were inhibited by the Chk1 kinase inhibitor UCN-01. Although not in itself toxic, UCN-01 increased the cytotoxicity of TMZ 5-fold, primarily by inhibiting cellular senescence and increasing the percentage of cells bypassing G(2)-M arrest and undergoing mitotic catastrophe. In addition to enhancing TMZ-induced cytotoxicity in p53-proficient cells, UCN-01 also blocked TMZ-induced Chk1 activation and transient G(2)-M arrest in p53-deficient U87MG-E6 cells and similarly enhanced TMZ-induced mitotic catastrophe and cell death. Taken together, these results indicate that Chk1 links TMZ-induced MMR to G(2)-M arrest. Furthermore, inhibition of the cytoprotective G(2) arrest pathway sensitizes cells to TMZ induced cytotoxicity and may represent a novel, mechanism-based means of increasing TMZ efficacy in both p53 wild-type and p53 mutant glioma cells. PMID- 11479225 TI - Heterogeneous T-cell response to MAGE-A10(254-262): high avidity-specific cytolytic T lymphocytes show superior antitumor activity. AB - MAGE-encoded antigens, which are expressed by tumors of many histological types but not in normal tissues, are suitable candidates for vaccine-based immunotherapy of cancers. Thus far, however, T-cell responses to MAGE antigens have been detected only occasionally in cancer patients. In contrast, by using HLA/peptide fluorescent tetramers, we have observed recently that CD8(+) T cells specific for peptide MAGE-A10(254-262) can be detected frequently in peptide stimulated peripheral blood mononuclear cells from HLA-A2-expressing melanoma patients and healthy donors. On the basis of these results, antitumoral vaccination trials using peptide MAGE-A10(254-262) have been implemented recently. In the present study, we have characterized MAGE-A10(254-262)-specific CD8(+) T cells in polyclonal cultures and at the clonal level. The results indicate that the repertoire of MAGE-A10(254-262)-specific CD8(+) T cells is diverse both in terms of clonal composition, efficiency of peptide recognition, and tumor-specific lytic activity. Importantly, only CD8(+) T cells able to recognize the antigenic peptide with high efficiency are able to lyse MAGE-A10 expressing tumor cells. Under defined experimental conditions, the tetramer staining intensity exhibited by MAGE-A10(254-262)-specific CD8(+) T cells correlates with efficiency of peptide recognition so that "high" and "low" avidity cells can be separated by FACS. Altogether, the data reported here provide evidence for functional diversity of MAGE-A10(254-262)-specific T cells and will be instrumental for the monitoring of peptide MAGE-A10(254-262)-based clinical trials. PMID- 11479226 TI - Murine six-transmembrane epithelial antigen of the prostate, prostate stem cell antigen, and prostate-specific membrane antigen: prostate-specific cell-surface antigens highly expressed in prostate cancer of transgenic adenocarcinoma mouse prostate mice. AB - To identify genes that are differentially up-regulated in prostate cancer of transgenic adenocarcinoma mouse prostate (TRAMP) mice, we subtracted cDNA isolated from mouse kidney and spleen from cDNA isolated from TRAMP-C1 cells, a prostate tumor cell line derived from a TRAMP mouse. Using this strategy, cDNA clones that were homologous to human six-transmembrane epithelial antigen of the prostate (STEAP) and prostate stem cell antigen (PSCA) were isolated. Mouse STEAP (mSteap) is 80% homologous to human STEAP at both the nucleotide and amino acid levels and contains six potential membrane-spanning regions similar to human STEAP. Mouse PSCA (mPsca) shares 65% homology with human PSCA at the nucleotide and amino acid levels. mRNA expression of mSteap and mPsca is largely prostate specific and highly detected in primary prostate tumors and metastases of TRAMP mice. Both mSteap and mPsca map to chromosome 5. Another known gene coding for mouse prostate-specific membrane antigen (mPsma) is also highly expressed in both primary and metastatic lesions of TRAMP mice. These results indicate that the TRAMP mouse model can be used to effectively identify genes homologous to human prostate-specific genes, thereby allowing for the investigation of their functional roles in prostate cancer. mSteap, mPsca, and mPsma constitute new tools for preventative and/or therapeutic vaccine construction and immune monitoring in the TRAMP mouse model that may provide insights into the treatment of human prostate cancer. PMID- 11479227 TI - Osteopontin is an autocrine mediator of hepatocyte growth factor-induced invasive growth. AB - In epithelial cells, hepatocyte growth factor (HGF) activates a genetic program involving cell-cell dissociation ("scattering"), growth and invasiveness. The full program is not elicited by other growth factors like epidermal growth factor, and is aberrantly activated during cancer progression to the invasive metastatic phenotype. To identify genes involved in the onset of invasive growth, we explored by cDNA microarrays the in vitro transcriptional response to HGF of mouse embryo liver cells. We identified osteopontin (OPN), a secreted matrix protein, as a major HGF transcriptional target. The wave of OPN induction is maximal at 6 h, in concomitance with the initiation of scattering, and is specific, because no other matrix protein among those explored by the microarray is affected. Interestingly, HGF, but not epidermal growth factor, promotes cell adhesion to OPN via the CD44 receptor. Scattering is significantly impaired by antibodies against OPN and CD44; conversely, constitutive OPN overexpression dramatically increases the motile and invasive responses to HGF, leading to disruption of the ordered morphogenetic program triggered by this ligand. PMID- 11479228 TI - Short polyglutamine tracts in the androgen receptor are protective against breast cancer in the general population. AB - We studied the association of breast cancer with the polymorphic polyglutamine repeat of the androgen receptor (AR) in 255 incident cases of breast cancer and 461 matched controls from the Quebec City metropolitan area. Women for whom the sum of both of the AR (CAG)n-repeats alleles is 39 or less (short-allele AR genotypes) have one-half the risk of breast cancer compared with women for whom the sum of AR (CAG)n-repeats is 40 or more [odds ratio (OR), 0.5; 95% confidence interval (CI), 0.3-0.83; P = 0.007]. This association is stronger in postmenopausal women (180 cases, 297 controls; OR, 0.36; 95% CI, 0.19-0.7; P = 0.003). We also observed an interaction between the type of menopause (natural versus surgical) and the AR genotype on breast cancer risk. Alternately, when subjects were grouped according to their (CAG)n-repeat genotype [homozygous for short alleles (CAG)n < or = 20; other genotypes ("long allele")], results were similar (OR. 0.5; 95% CI, 0.27-0.82; P = 0.007). Thus, women with short-alleles AR genotypes appear to be protected against breast cancer. Short-alleles AR genotypes were observed in 16% of the general population as represented by the control group. Short polyglutamine repeats in the AR protein have been reported to be associated with an increase in the capacity of the receptor to activate transcription of reporter genes in vitro. Furthermore, androgens have been previously shown to inhibit in vitro the growth of breast cancer cell lines. This suggests that differences in the number of polyglutamines in the AR protein may influence individual risk of breast cancer, especially in postmenopausal women, and that this apparent protection could be the consequence of an increased response/sensitivity to androgens. PMID- 11479229 TI - Global and gene-specific epigenetic patterns in human bladder cancer genomes are relatively stable in vivo and in vitro over time. AB - We used a methylation-sensitive arbitrarily primed PCR technique to analyze, in a nonselective manner, methylation alterations at GC-rich regions of the genome in metachronous tumors and their derived cell lines from two patients with transitional cell carcinoma of the bladder. The methylation status of the majority of evaluable sequences (83%) remained unchanged in the tumors from both patients relative to a panel of normal urothelium samples obtained from individuals free of bladder disease, in which we measured <1% interindividual variation. The 17% of methylation alterations represents sequences altered in either a cancer-specific (3%), tumor-specific (1%), or patient-specific (13%) manner. The proportion of the altered sequences analyzed that were CpG islands corresponds to approximately 7000 CpG islands altered in the genome. Surprisingly, few additional changes in methylation patterns were observed in cell lines derived from the tumors; however, all of the cell lines showed altered methylation in a common set of 3% of evaluable sequences. Three genes known to be aberrantly methylated in bladder cancer (p16, p15, and PAX6) were studied in detail by methylation-sensitive single nucleotide primer extension and showed increased methylation in culture at preexisting methylated sites for all of the exons but no de novo methylation in culture for the promoters in any cell line. Therefore, our investigation provides the first serial as well as parallel quantitation of the global epigenetic stability in two independent bladder cancer genomes over the course of progression and in culture. In addition, our investigation also provides the first direct comparison of the epigenetic and genetic patterns on the global scale, showing the epigenetic pattern to be relatively stable in vivo and in vitro over time within an individual. PMID- 11479230 TI - Regulation of the catalase gene promoter by Sp1, CCAAT-recognizing factors, and a WT1/Egr-related factor in hydrogen peroxide-resistant HP100 cells. AB - Reactive oxygen species play a critical role in the onset of apoptosis induced by various extracellular stimuli, including ionizing radiation. Therefore active regulation of reactive oxygen species-metabolizing enzymes may be one response to an apoptotic stimulus. In this regard, HP100 cells, H(2)O(2)-resistant variants derived from human leukemia HL60 cells, display an interesting phenotype in which the activity of catalase is constitutively high, whereas its mRNA is reduced after X-ray irradiation. In the present study, we investigated the molecular mechanisms underlying this phenomenon. By combining analyses from nuclear run-on, reporter gene transient transfection, genomic footprinting, site-directed mutagenesis, electrophoretic mobility shift analysis, and Western blotting experiments, we found that constitutively elevated catalase expression is strongly regulated at the transcriptional level by both Sp1 and CCAAT-recognizing factors and that much higher levels of nuclear Sp1 and NF-Y are present in HP100 nuclei as compared with HL60 nuclei. In addition, we demonstrated an X-ray inducible association of a WT1/Egr-related factor with an overlapping Sp1/Egr-1 recognition sequence located within the core promoter of the catalase gene. This association may lead to inactivation of the promoter by disturbing or competing with the transactivating ability of Sp1. PMID- 11479231 TI - Genetic analysis of early- versus late-stage ovarian tumors. AB - In the United States, ovarian cancer is the fourth most common cause of cancer related deaths among women. The most important prognostic factor for this cancer is tumor stage, or extent of disease at diagnosis. Although women with low-stage tumors have a relatively good prognosis, most women diagnosed with late-stage disease eventually succumb to their cancer. In an attempt to understand early events in ovarian carcinogenesis, and to explore steps in its progression, we have applied multiple molecular genetic techniques to the analysis of 21 early stage (stage I/II) and 17 advanced-stage (stage III/IV) ovarian tumors. These techniques included expression profiling with cDNA microarrays containing approximately 18,000 expressed sequences, and comparative genomic hybridization to address the chromosomal locations of copy number gains as well as losses. Results from the analysis indicate that early-stage ovarian cancers exhibit profound alterations in gene expression, many of which are similar to those identified in late-stage tumors. However, differences observed at the genomic level suggest differences between the early- and late-stage tumors and provide support for a progression model for ovarian cancer development. PMID- 11479232 TI - Tumor suppressor genes in the 9p21 gene cluster are selective targets of inactivation in neuroendocrine gastroenteropancreatic tumors. AB - Functional inactivation of the Rb and p53 pathways appears to be a rite of passage for all cancerous cells. However, p53 and Rb alterations are rare events in neuroendocrine gastroenteropancreatic (GEP) tumors. The CDKN2 locus on chromosome 9p21 sits at the nexus of both pathways harboring tumor suppressor genes, which restrain cell growth by affecting the function of pRb and p53. Therefore, we analyzed the implication of their inactivation in 37 primary neuroendocrine GEP tumors and two cell culture models. RT-PCR analysis revealed loss of expression of at least one of the tumor suppressor genes CDKN2A/p16, CDKN2B/p15, and CDKN2D/p14 with distinct genetic profiles, most frequently in nonfunctional pancreatic tumors (57%) and small intestinal carcinoids (44%), and less commonly in insulinomas (30%) and gastrinomas (22%). DNA analysis and methylation-specific PCR attributed loss of expression to either homozygous deletion or 5'CpG island hypermethylation. 5-Aza-2-deoxycytidine treatment reversed CDKN2A/p16 and CDKN2B/p15 silencing with concurrent growth restraint. Thus, tumor suppressor genes localized in the 9p21 gene cluster are specific targets of inactivation in neuroendocrine GEP tumors, and demethylating agents might hold promise for selective therapy. PMID- 11479233 TI - Hepatocyte growth factor/scatter factor-induced activation of MEK and PI3K signal pathways contributes to expression of proangiogenic cytokines interleukin-8 and vascular endothelial growth factor in head and neck squamous cell carcinoma. AB - The proangiogenic activity of hepatocyte growth factor (HGF)/scatter factor has been closely associated with its ability to stimulate endothelial cell chemotaxis, migration, proliferation, and capillary formation. However, the potential of HGF as a paracrine factor in regulating the expression of angiogenesis factors by tumor cells is not widely appreciated. We observed that increased HGF was correlated with higher levels of angiogenesis factors interleukin (IL)-8 and vascular endothelial growth factor (VEGF) in serum of patients with head and neck squamous cell carcinoma (HNSCC) as compared with that in normal volunteers and hypothesized that HGF may regulate angiogenesis factor production by tumor cells through the activation of its receptor c-Met, which is expressed by HNSCC cells. To test this hypothesis, we examined the effect of HGF treatment on IL-8 and VEGF expression by a panel of primary keratinocytes and HNSCC lines. HGF induced a significant dose-dependent increase in IL-8 and/or VEGF cytokine production in eight HNSCC lines tested, which is not observed in normal keratinocytes. In addition, HGF increased mRNA expression of IL-8 in 3 of 6 and VEGF in 5 of 6 HNSCC lines. The increase in induction of these factors by HGF corresponded to an increase in phosphorylation of c-Met in HNSCC. HGF-induced phosphorylation of mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) pathway substrate p42/p44(erk) and phosphatidylinositol 3' kinase (PI3K) pathway substrate Akt provided evidence for downstream activation of MEK and PI3K pathways in HNSCC. Inhibitors of MEK (U0126) and PI3K (LY294002) blocked p42/p44(erk) and Akt, respectively, and partially blocked HGF-induced production of IL-8 and VEGF, whereas the combination of U0126 and LY294002 completely inhibited expression of IL-8 and VEGF by UMSCC-11A. Our results demonstrate that HGF can promote expression of angiogenesis factors in tumor cells through both MEK- and PI3K-dependent pathways. Understanding HGF/Met paracrine regulatory mechanisms between tumor and host cells may provide critical information for targeting of therapies against angiogenesis. PMID- 11479234 TI - Abrogation of transforming growth factor-alpha/epidermal growth factor receptor autocrine signaling by an RXR-selective retinoid (LGD1069, Targretin) in head and neck cancer cell lines. AB - Clinical studies have demonstrated that retinoids, including retinol (Vitamin A) and its synthetic derivatives, can eradicate leukoplakia and suppress the formation of squamous cell carcinoma of the head and neck (SCCHN). Nonselective retinoids have been shown to abrogate transcriptional activation of transforming growth factor-alpha (TGF-alpha) and epidermal growth factor receptor (EGFR), which characterize SCCHN. LGD1069 (Targretin) is a potent RXR-selective retinoic acid agonist with a reduced toxicity profile compared with other nonselective retinoids. We examined the effect of LGD1069 (10 microm) on cellular proliferation and expression of putative intermediate biomarker genes including TGF-alpha, EGFR, and RAR-beta in seven SCCHN cell lines. A quantitative reverse transcription-PCR assay using a novel "primer dropping" method was used to determine expression levels of EGFR, TGF-alpha, and RAR-beta before and after treatment with LGD1069 (10 microM). SCCHN proliferation was reduced by a mean of 50% at 4 days in seven SCCHN cell lines after LGD1069 treatment (P < or = 0.05). EGFR expression levels were decreased by a mean of 58.4% (P = 0.007), TGF-alpha levels were decreased by a mean of 28.8% (P = 0.01), and RAR-beta levels were increased by a mean of 60% (P = 0.03). TGF-alpha stimulation of EGFR is associated with constitutive signal transducer and activator of transcription 3 (Stat3) activation in SCCHN. Abrogation of constitutive Stat3 activation was seen with LGD1069 treatment. These results suggest that an RXR-selective retinoic acid decreases SCCHN proliferation in part by interfering with TGF-alpha/EGFR autocrine signaling. PMID- 11479235 TI - Ubiquitin/26S proteasome-mediated degradation of topoisomerase I as a resistance mechanism to camptothecin in tumor cells. AB - Camptothecin (CPT) induces down-regulation of topoisomerase I (TOP1) via an ubiquitin/26S proteasome pathway. Studies using a panel of breast and colorectal cancer cell lines as well as primary nontransformed and oncogene-transformed cells have demonstrated that CPT-induced down-regulation exhibits a high degree of heterogeneity. In general, nontransformed cells are much more proficient in CPT-induced TOP1 down-regulation than their transformed counterparts. Among the breast and colorectal cancer cell lines, there was a general correlation between the extent of CPT-induced TOP1 down-regulation and CPT resistance. The breast cancer cell line ZR-75-1, the most sensitive to CPT, was completely defective in CPT-induced TOP1 down-regulation, whereas the breast cancer cell line BT474, the least sensitive to CPT, exhibited effective CPT-induced TOP1 down-regulation. The 26S proteasome inhibitor MG132 was shown to inhibit CPT-induced down-regulation of TOP1 in BT474 cells and selectively sensitized BT474 but not ZR-75-1 cells to CPT-induced cytotoxicity and apoptosis. In the aggregate, these results suggest that CPT-induced down-regulation of TOP1 could be an important parameter for determining CPT sensitivity/resistance in tumor cells. Analysis of the levels of TOP1 cleavable complexes, SUMO-1-TOP1 conjugates, and ubiquitin-TOP1 conjugates in ZR-75-1 and BT474 cells has suggested that the heterogeneity of CPT-induced down-regulation of TOP1 in tumor cells is at least in part attributable to altered regulation of a process(es) downstream from the TOP1 cleavable complex. PMID- 11479236 TI - Signaling from protease-activated receptor-1 inhibits migration and invasion of breast cancer cells. AB - Proteases give cancer a defining characteristic of being able to break through extracellular matrix barriers and invade into other tissues in response to chemotactic signals. Recently, the cell surface protease-activated receptor (PAR) 1 has been shown to act as a chemokine receptor in inflammatory cells, and its expression is tightly correlated with metastatic propensity of breast cancer cells. The aim of the present study was to determine whether activation of PAR1 or the other known PARs (PAR2-4) can regulate migration and invasion of breast cancer cells. We found that the highly invasive MDAMB231 breast cancer cell line expressed very high levels of functional PAR1, PAR2, and PAR4, whereas minimally invasive MCF7 cells had trace amounts of PAR1 and low levels of PAR2 and PAR4. Despite the differences in expression, PAR2 and PAR4 acted as chemokine receptors in both invasive and minimally invasive breast cell lines. Quite unexpectedly, we found that activation of PAR1 with thrombin or the peptide agonist SFLLRN markedly inhibited invasion and migration of MDAMB231 cells when applied as a concentration gradient in the direction of cell movement. Additionally, we demonstrated that inhibition of chemotaxis was mediated through a G(i)/phosphoinositide-3-OH kinase-dependent pathway. Activation of G(i) signaling with epinephrine or wasp venom mastoparan also inhibited invasion and migration of the breast cancer cells. These findings suggest that therapeutics targeted toward G(i)-couplers that are selectively expressed in breast cancer cells could prove beneficial in halting the progression of invasive breast cancer. PMID- 11479237 TI - Efficient replication of adenovirus despite the overexpression of active and nondegradable p53. AB - The adenoviral oncoproteins E1B-55 kDa and E4orf6 inactivate and destabilize the tumor suppressor protein p53, thereby contributing to malignant transformation. However, it is unclear whether the elimination of p53 also contributes to the efficiency of viral replication. Furthermore, it is controversial whether adenoviruses with a deletion in the E1B-55 kDa-coding region might selectively replicate in cells with a mutation or deletion of the p53 gene and, therefore, represent a tool in cancer therapy. To address the role of p53 in virus replication, amino acid substitutions were introduced into the NH(2)-terminal portion of p53, replacing residues 24-28 with the corresponding sequence of the human p53-homologue p73. This replacement leaves p53 transcriptionally active but renders the modified protein, termed p53mt24-28, completely resistant to inhibition and degradation by adenoviral oncoproteins. Surprisingly, even strong overexpression of p53 or p53mt24-28 allowed the virus to replicate as efficiently as in the absence of p53 proteins, both in tumor cells and in primary endothelial cells. Also, p53 or p53mt24-28 did not reduce the amount of virus released from infected cells. These observations were made in primary cells or in cell lines that were capable of expressing the p53-agonist p14ARF. Thus, active p53 does not inhibit the growth of adenovirus. Alternative strategies should be used to improve the utility of adenoviruses in cancer therapy. PMID- 11479238 TI - Correspondence re: A. Rothfuss et al., Induced micronucleus frequencies in peripheral blood lymphocytes as a screening test for carriers of a BRCA1 mutation in breast cancer families. Cancer Res., 60: 390-394, 2000. PMID- 11479239 TI - Correspondence re: P. P. Claudio et al., Mutations in the retinoblastoma-related gene RB2/p130 in primary nasopharyngeal carcinoma. Cancer Res., 60: 8-12, 2000. PMID- 11479241 TI - V. Craig Jordan, Ph.D., D.Sc., receives Bristol-Myers Squibb Award. PMID- 11479244 TI - Who's on first? Does hypertension in type I diabetics result from nephropathy or metabolic alterations? PMID- 11479245 TI - Nebivolol reverses endothelial dysfunction in essential hypertension: a randomized, double-blind, crossover study. AB - BACKGROUND: Vascular endothelial dysfunction may predict future atherosclerosis. Hence, an antihypertensive agent that reverses endothelial dysfunction and lowers blood pressure might improve the prognosis of patients with hypertension. We hypothesized that nebivolol, a vasodilating beta-blocker, could improve endothelial dysfunction. We tested this hypothesis by comparing the effects of nebivolol and atenolol on endothelial function. METHODS AND RESULTS: Twelve hypertensive patients with a mean ambulatory blood pressure of 154+/-7/97+/-10 mm Hg were randomized after a 2-week placebo run-in period (baseline) in a double blind, crossover fashion to 8-week treatment periods with either 5 mg of nebivolol with 2.5 mg of bendrofluazide or 50 mg of atenolol with 2.5 mg of bendrofluazide. Forearm venous occlusion plethysmography and intra-arterial infusions of acetylcholine and N(G)-monomethyl-L-arginine (L-NMMA) were used to assess stimulated and basal endothelium-dependent nitric oxide release, respectively. Sodium nitroprusside was used as an endothelium-independent control. Nebivolol/bendrofluazide and atenolol/bendrofluazide each lowered the clinic blood pressure to the same extent (132+/-7/82+/-6 and 132+/-9/83+/-8 mm Hg, respectively; P<0.001 from baseline). The vasodilatory response to acetylcholine was significantly increased with nebivolol/bendrofluazide (maximum percentage change in forearm blood flow [mean+/-SEM], 435+/-27%, P<0.001) but not with atenolol/bendrofluazide. Similarly, the endothelium-dependent vasoconstrictive response to L-NMMA was significantly improved only with nebivolol treatment (percentage change in forearm blood flow, -54+/-5%; P<0.001). The response to sodium nitroprusside was not different between treatments, suggesting that the endothelium-independent pathway was unaffected. CONCLUSIONS: Nebivolol/bendrofluazide increased both stimulated and basal endothelial nitric oxide release, whereas for the same degree of blood pressure control, atenolol/bendrofluazide had no effect on nitric oxide bioactivity. Thus, nebivolol may offer additional vascular protection in treating hypertension. PMID- 11479246 TI - Prospective multicenter study of pregnancy outcomes in women with heart disease. AB - BACKGROUND: The maternal and neonatal risks associated with pregnancy in women with heart disease receiving comprehensive prenatal care have not been well defined. METHODS AND RESULTS: We prospectively enrolled 562 consecutive pregnant women with heart disease and determined the outcomes of 599 pregnancies not ending in miscarriage. Pulmonary edema, arrhythmia, stroke, or cardiac death complicated 13% of pregnancies. Prior cardiac events or arrhythmia, poor functional class or cyanosis, left heart obstruction, and left ventricular systolic dysfunction independently predicted maternal cardiac complications; the cardiac event rate can be predicted using a risk index incorporating these predictors. Neonatal complications (20% of pregnancies) were associated with poor functional class or cyanosis, left heart obstruction, anticoagulation, smoking, and multiple gestations. CONCLUSIONS: Pregnancy in women with heart disease is associated with significant cardiac and neonatal complications, despite state-of the-art obstetric and cardiac care. Maternal cardiac risk can be predicted with the use of a risk index. PMID- 11479247 TI - Effects of ramipril on coronary events in high-risk persons: results of the Heart Outcomes Prevention Evaluation Study. AB - BACKGROUND: In trials of patients with left ventricular dysfunction or heart failure, ACE inhibitor use was unexpectedly associated with reduced myocardial infarction (MI). Using the Heart Outcomes Prevention Evaluation (HOPE) trial data, we tested prospectively whether ramipril, an ACE inhibitor, could reduce coronary events and revascularization procedures among patients with normal left ventricular function. METHODS AND RESULTS: In the HOPE trial, 9297 high-risk men and women, >/=55 years of age with previous cardiovascular disease or diabetes plus 1 risk factor, were randomly assigned to ramipril (up to 10 mg/d), vitamin E (400 IU/d), their combination, or matching placebos. During the mean follow-up of 4.5 years, there were 482 (10.4%) patients with clinical MI and unexpected cardiovascular death in the ramipril group compared with 604 (12.9%) in the placebo group [relative risk reduction (RRR), 21% (95% CI) (11,30); P<0.0003]. Ramipril was associated with a trend toward less fatal MI and unexpected death [4.0% versus 4.7%; RRR, 16% (-3, 31)] and with a significant reduction in nonfatal MI [5.6% versus 7.2%; RRR, 23% (9,34)]. Risk reductions in MI were documented in participants taking or not taking beta-blockers, lipid lowering, and/or antiplatelet agents. Although ramipril had no impact on hospitalizations for unstable angina [11.9% versus 12.2%; RRR, 3% (-9,14)], it reduced the risk of worsening and new angina [27.2% versus 30.0%; RRR, 12% (5,18); P<0.0014] and coronary revascularizations [12.5% versus 14.8%; RRR, 18%; (8,26) P<0.0005]. CONCLUSIONS: In this high-risk cohort, ramipril reduced the risk of MI, worsening and new angina, and the occurrence of coronary revascularizations. PMID- 11479248 TI - Abnormal longitudinal, base-to-apex myocardial perfusion gradient by quantitative blood flow measurements in patients with coronary risk factors. AB - BACKGROUND: A longitudinal, base-to-apex myocardial perfusion gradient has been described in patients with coronary artery disease (CAD) and was attributed to diffuse coronary luminal narrowing. We asked whether an abnormal perfusion gradient also existed in patients without CAD but with coronary risk factors. We measured myocardial blood flow (MBF) with (13)N-ammonia and PET at rest and during hyperemia in patients with coronary risk factors but without CAD. METHODS AND RESULTS: Regional MBF was measured in absolute units with (13)N-ammonia and PET at rest and during dipyridamole hyperemia in 36 patients with coronary risk factors (age, 55+/-10 years) and in 36 age-matched (age, 53+/-10 years) and in 28 young (age, 25+/-5 years) normal subjects. MBF was determined globally, for each of the 3 coronary territories, and in the mid and mid-to-apical sections of the left ventricle (LV). Myocardial perfusion on qualitative analysis was normal at rest and during hyperemia, and no flow defects were present. MBF in absolute units was similar in the 3 coronary territories. However, hyperemic MBFs in the mid-to-apical LV section were lower than in the mid LV section in the "at-risk" group (2.04+/-0.61 versus 1.71+/-0.40 mL. min(-1). g(-1); P<0.004) but not in the age-matched or in the young normal subjects. CONCLUSIONS: The abnormal longitudinal, base-to-apex perfusion gradient observed during dipyridamole MBF suggests the presence of a functional and/or structural alteration of the coronary circulation associated with coronary risk factors, possibly reflecting developing coronary atherosclerosis or preclinical CAD. PMID- 11479249 TI - Clinical and economic impact of diabetes mellitus on percutaneous and surgical treatment of multivessel coronary disease patients: insights from the Arterial Revascularization Therapy Study (ARTS) trial. AB - BACKGROUND: Our aims were to compare coronary artery bypass grafting (CABG) and stenting for the treatment of diabetic patients with multivessel coronary disease enrolled in the Arterial Revascularization Therapy Study (ARTS) trial and to determine the costs of these 2 treatment strategies. METHODS AND RESULTS: Patients (n=1205) were randomly assigned to stent implantation (n=600; diabetic, 112) or CABG (n=605; diabetic, 96). Costs per patient were calculated as the product of each patient's use of resources and the corresponding unit costs. Baseline characteristics were similar between the groups. At 1 year, diabetic patients treated with stenting had the lowest event-free survival rate (63.4%) because of a higher incidence of repeat revascularization compared with both diabetic patients treated with CABG (84.4%, P<0.001) and nondiabetic patients treated with stents (76.2%, P=0.04). Conversely, diabetic and nondiabetic patients experienced similar 1-year event-free survival rates when treated with CABG (84.4% and 88.4%). The total 1-year costs for stenting and CABG in diabetic patients were $12 855 and $16 585 (P<0.001) and in the nondiabetic groups, $10 164 for stenting and $13 082 for surgery. CONCLUSIONS: Multivessel diabetic patients treated with stenting had a worse 1-year outcome than patients assigned to CABG or nondiabetics treated with stenting. The strategy of stenting was less costly than CABG, however, regardless of diabetic status. PMID- 11479250 TI - Randomized comparison of ticlopidine and clopidogrel after intracoronary stent implantation in a broad patient population. AB - BACKGROUND: Although clopidogrel is used to prevent subacute stent thrombosis, its safety and efficacy have not been compared with ticlopidine in a randomized manner in the United States. METHODS AND RESULTS: Patients with successful intracoronary stent implantation were randomly assigned to therapy with ticlopidine or clopidogrel. Loading doses were administered immediately after the procedure, and the drugs were prescribed for 2 weeks. One thousand sixteen patients were enrolled: 522 patients were randomly assigned to ticlopidine therapy and 494 to clopidogrel. High-risk characteristics included recent myocardial infarction in 41.4% of the cases, angiographically evident thrombus in 20.9%, and abrupt or threatened closure in 3.64%. An intravenous glycoprotein IIb/IIIa inhibitor was used in 48.2% of the cases, and thrombocytopenia occurred in 1.43% of these patients. Failure to complete 2 weeks of therapy occurred in 3.64% of the patients treated with ticlopidine and in 1.62% of the patients treated with clopidogrel (P=0.043). Within 30 days, thrombosis of the stent occurred in 1.92% of the patients in the ticlopidine group and in 2.02% of the clopidogrel group (P=0.901). A major adverse cardiac event occurred in 4.60% of patients receiving ticlopidine and in 3.85% of patients receiving clopidogrel (P=0.551). CONCLUSIONS: Clopidogrel is better tolerated than ticlopidine during a 2-week regimen after intracoronary stent implantation. Combining either thienopyridine with an intravenous platelet IIb/IIIa inhibitor appears to be safe. When applied to a broad spectrum of patients receiving stent implantation, clopidogrel confers similar protection as ticlopidine against subacute stent thrombosis and major adverse cardiac events. PMID- 11479251 TI - Peripheral chemoreceptor hypersensitivity: an ominous sign in patients with chronic heart failure. AB - BACKGROUND: Peripheral chemoreceptor hypersensitivity is a feature of abnormal cardiorespiratory reflex control in chronic heart failure (CHF) and may contribute to sympathetic overactivity, attenuated baroreflex sensitivity (BRS), and excessive ventilation during exercise. We studied whether augmented peripheral chemosensitivity carries independent prognostic significance. METHODS AND RESULTS: We assessed peripheral chemosensitivity (ventilatory response to hypoxia using transient inhalation of pure nitrogen) and BRS (phenylephrine and spectral methods) in 80 consecutive CHF patients (age 58+/-9 years; left ventricular ejection fraction [LVEF] 24+/-12%; peak oxygen consumption [peak VO(2)] 18+/-7 mL(-1). min(-1)). CHF patients demonstrated augmented peripheral chemosensitivity and decreased BRS (all P<0.01 versus reference values). During follow-up (median 41 months, >3 years in all survivors), 37 patients died. High peripheral chemosensitivity (>0.72 L. min(-1). %SaO(2)(-1)) predicted impaired survival (hazard ratio 3.2, 95% CI 1.6 to 6.0, P=0.0006). In the 27 patients (34%) with high peripheral chemosensitivity, 3-year survival was 41% (95% CI 22% to 60%) compared with 77% (66% to 89%) in 53 patients with normal chemosensitivity (P=0.0002). In multivariate analyses, augmented chemosensitivity independently predicted death (hazard ratio 2.8, 95% CI 1.5 to 5.5, adjusted for age, peak VO(2), and VE/VCO(2) [P=0.002]; hazard ratio 2.6, 95% CI 1.3 to 5.1, adjusted for age, LVEF, and peak VO(2) [P=0.008]). Depressed BRS was related to unfavorable prognosis in univariate analysis (P=0.05) but not in multivariate analyses. CONCLUSIONS: Hypersensitivity of the peripheral chemoreceptors independently predicts adverse prognosis in ambulatory patients with CHF. This hyperactive excitatory reflex, through its inhibitory effect on the baroreflex, may be the reason for the previously observed prognostic association of the latter. PMID- 11479252 TI - Reversing the direction of paced ventricular and atrial wavefronts reveals an oblique course in accessory AV pathways and improves localization for catheter ablation. AB - BACKGROUND: The purpose of this study was to determine how often accessory atrioventricular (AV) pathways (AP) cross the AV groove obliquely. With an oblique course, the local ventriculoatrial (VA) interval at the site of earliest atrial activation (local-VA) and the local-AV interval at the site of earliest ventricular activation (local-AV) should vary by reversing the direction of the paced ventricular and atrial wavefronts, respectively. METHODS AND RESULTS: One hundred fourteen patients with a single AP were studied. Two ventricular and two atrial pacing sites on opposite sides of the AP were selected to reverse the direction of the ventricular and atrial wavefronts along the annulus. Reversing the ventricular wavefront increased local-VA by >/=15 ms in 91 of 106 (91%) patients. With the shorter local-VA, the ventricular potential overlapped the atrial potential along a 17.2+/-8.5-mm length of the annulus. No overlap occurred with the opposite wavefront. Reversing the atrial wavefront increased local-AV by >/=15 ms in 32 of 44 (73%) patients. With the shorter local-AV, the atrial potential overlapped the ventricular potential along an 11.9+/-8.9-mm length of the annulus. No overlap occurred with the opposite wavefront. Mapping during longer local-VA or local-AV identified an AP potential in 102 of 114 (89%) patients. Catheter ablation eliminated AP conduction in all 111 patients attempted (median, 1 radiofrequency application in 99 patients with an AP potential versus 4.5 applications without an AP potential). CONCLUSIONS: Reversing the direction of the paced ventricular or atrial wavefront reveals an oblique course in most APs and facilitates localization of the AP potential for catheter ablation. PMID- 11479253 TI - Clinical implications for affected parents and siblings of probands with long-QT syndrome. AB - BACKGROUND: Whenever a proband is identified with long-QT syndrome (LQTS), his or her parents and siblings should be evaluated regarding the possibility of carrying the disorder. In the majority of cases, one of the proband's parents and one or more siblings are affected. The aim of this study was (1) to determine whether the clinical severity of LQTS in the proband is useful in identifying first-degree family members at high risk for cardiac events, and (2) to evaluate the clinical course of affected parents and siblings of LQTS probands. METHODS AND RESULTS: The clinical and ECG characteristics of 211 LQTS probands and 791 first-degree relatives (422 parents and 369 siblings) were studied to determine if the clinical profile of the proband is useful in determining the clinical severity of LQTS in affected parents and siblings. Affected female parents of an LQTS proband had a greater cumulative risk for a first cardiac event than affected male parents. The probability of a parent or sibling having a first cardiac event was not significantly influenced by the severity of the proband's clinical symptoms. Female sex and QT(c) duration were risk factors for cardiac events among affected parents, and QT(c) was the only risk factor for cardiac events in affected siblings. CONCLUSIONS: The severity profile of LQTS in a proband was not found to be useful in identifying the clinical severity of LQTS in affected first-degree relatives of the proband. PMID- 11479254 TI - Improvement in hypertension in patients with diabetes mellitus after kidney/pancreas transplantation. AB - BACKGROUND: Hypertension persists in many patients with diabetes mellitus after kidney transplantation. However, the impact of control of diabetes as well as kidney failure on hypertension by combined kidney and pancreas transplantation has not been studied. METHODS AND RESULTS: Between March 1993 and August 1998, 111 patients with type 1 diabetes mellitus underwent successful pancreas transplantation (108 kidney/pancreas transplantation) and another 28 patients with type 1 diabetes mellitus underwent isolated kidney transplantation. Blood pressure measurements and all antihypertensive medications were determined for both groups before transplantation and at 1, 3, 6, and 12 months and at the most recent outpatient evaluation after transplantation. At baseline, the mean blood pressure was 151/88 and 151/83 mm Hg for the kidney/pancreas and isolated kidney transplant patients, respectively. The mean blood pressure decreased to 134/77 mm Hg 1 month after kidney/pancreas transplantation (P<0.001) and decreased further to 126/70 mm Hg (P<0.001) at a mean follow-up of 18 months. This reduction in blood pressure after transplantation occurred despite a decrease in antihypertensive medications and the institution of immunosuppressive agents. At 1 month after kidney/pancreas transplantation, the average number of antihypertensive medications per patient was 0.9+/-1.0, compared with 2.5+/-1.1 before surgery (P<0.001). At 18 months after transplantation, 34% of patients were both normotensive (blood pressure > galactose) and the effects of ligand valency (tetraantennary > triantennary >> diantennary >> monoantennary) and sugar spacing (20 A 10 A 4 A) are well documented, the effect of particle size on recognition and uptake of ligands by the receptor is poorly defined. In the present study, we assessed the maximum ligand size that still allows effective processing by the ASGPr of mouse hepatocytes in vivo and in vitro. Here too, we synthesized a novel glycolipid, which possesses a highly hydrophobic steroid moiety for stable incorporation into liposomes, and a triantennary GalNAc(3)-terminated cluster glycoside with a high nanomolar affinity (2 nm) for the ASGPr. Incorporation of the glycolipid into small (30 nm) [(3)H]cholesteryl oleate-labeled long circulating liposomes (1-50%, w/w) caused a concentration-dependent increase in particle clearance that was liver-specific (reaching 85 +/- 7% of the injected dose at 30 min after injection) and mediated by the ASGPr on hepatocytes, as shown by competition studies with asialoorosomucoid in vivo. By using glycolipid laden liposomes of various sizes between 30 and 90 nm, it was demonstrated that particles with a diameter of >70 nm could no longer be recognized and processed by the ASGPr in vivo. This threshold size for effective uptake was not related to the physical barrier raised by the fenestrated sinusoidal endothelium, which shields hepatocytes from the circulation, because similar results were obtained by studying the uptake of liposomes on isolated mouse hepatocytes in vitro. From these data we conclude that in addition to the species, valency, and orientation of sugar residues, size is also an important determinant for effective recognition and processing of substrates by the ASGPr. Therefore, these data have important implications for the design of ASGPr-specific carriers that are aimed at hepatocyte-directed delivery of drugs and genes. PMID- 11479286 TI - Molecular and cellular physiology of the dissociation of atrial natriuretic peptide from guanylyl cyclase a receptors. AB - Guanylyl cyclase subtype A (GCA) is the main receptor that mediates the effects of atrial natriuretic peptide (ANP) in the regulation of plasma volume and blood pressure. The dynamics of the dissociation of ANP from GCA were investigated in cultured Chinese hamster ovary (CHO) cells stably transfected with wild-type (WT) or mutant GCA receptors. The rate of dissociation of specifically bound (125)I ANP-(1-28) from intact CHOGCAWT cells at 37 degrees C was extremely rapid (K(off) = 0.49 +/- 0.02 min(-1)), whereas in isolated membranes prepared from these cells, the dissociation at 37 degrees C was >10-fold slower (K(off) = 0.035 +/- 0.006 min(-1)). The dissociation of ANP from CHOGCAWT cells showed remarkable temperature dependence. Between 22 and 37 degrees C, K(off) increased approximately 8 times, whereas between 4 and 22 degrees C, it increased only 1.5 times. Total deletion of the cytoplasmic domain or of the catalytic guanylyl cyclase sequence within this domain abolished ANP-induced increases in cGMP, dramatically slowed receptor-ligand dissociation by at least 10-fold, and abolished the temperature dependence of the dissociation of ANP. Deletion of the kinase-like domain led to maximal constitutive activation of guanylyl cyclase, markedly decreased K(off) to 0.064 +/- 0.006 min(-1), and also abolished the temperature dependence of dissociation. Substitution of Ser(506) by Ala and particularly the double substitution of Gly(505) and Ser(506) by Ala within the kinase-like domain markedly reduced ANP-induced increases in cGMP, whereas K(off) decreased modestly (albeit significantly) to 0.36 +/- 0.03 and 0.24 +/- 0.02 min( 1), respectively. As a whole, the results demonstrate for the first time that temperature per se or ATP alone cannot account for rapid GCA receptor-ligand dissociation under physiological conditions and suggest that ligand dissociation is modulated in part by the interaction of still unidentified cytosolic factors with the cytoplasmic domain of GCA. PMID- 11479287 TI - The contribution of arginine residues within the P6-P1 region of alpha 1 antitrypsin to its reaction with furin. AB - A series of mutants incorporating furin recognition sequences within the P6-P1 region of the reactive site loop of alpha(1)-antitrypsin were constructed. Variants containing different combinations of basic residues in the P1, P2, P4, and P6 positions replacing the wild type (P6)LEAIPM(P1) sequence were evaluated for their capacity to establish SDS-resistant complexes with furin, to affect association rate constants (k(ass) and k'(ass)), or to inhibit furin-dependent proteolysis of a model precursor in vivo. Each variant abolished processing of pro-von Willebrand factor in transfected hEK293 cells. The k(ass) of all variants were found to be similar (1.1-1.7 x 10(6) m(-1) s(-1)) except for one mutant, RERIRR, which had a k(ass) of 3.3 x 10(5) m(-1) s(-1). However, the stoichiometry of inhibition varied with values ranging from 2.9 to >24, indicating rapid formation of the acyl-enzyme intermediate (high k'(ass)). Moreover, those variants having high stoichiometry of inhibition values were accompanied by the rapid formation of cleaved forms of the inhibitors. The data suggest that the rate of conversion of the acyl-enzyme (EI') into the highly stable complex (EI*) was affected by replacement of specific residues within the reactive site loop. Taken together, the results reveal how furin recognition sequences within the context of the biochemical properties of serpins will play a role in the capacity of the protein to follow either the inhibitory or the substrate pathway. PMID- 11479288 TI - Functional interaction of calcium-/calmodulin-dependent protein kinase II and cytosolic phospholipase A(2). AB - Calcium-/calmodulin-dependent protein kinase II (CaM kinase II), a decoder of Ca(2+) signals, and cytosolic phospholipase A(2) (cPLA(2)), an enzyme involved in arachidonate release, are involved in many physiological and pathophysiological processes. Activation of CaM kinase II in norepinephrine-stimulated vascular smooth muscle cells leads to activation of cPLA(2) and arachidonic acid release. Surface plasmon resonance, mass spectrometry, and kinetic studies show that CaM kinase II binds to cPLA(2) resulting in cPLA(2) phosphorylation on Ser-515 and an increase in its enzymatic activity. Phosphopeptide mapping studies with cPLA(2) from norepinephrine-stimulated smooth muscle cells indicates that phosphorylation of cPLA(2) on Ser-515, but not on Ser-505 or Ser-727, occurs in vivo. This novel signaling pathway for arachidonate release is shown to be cPLA(2)-dependent by use of a recently described and highly selective inhibitor of this enzyme. PMID- 11479289 TI - Caspase-mediated cleavage of the Ca2+/calmodulin-dependent protein kinase-like kinase facilitates neuronal apoptosis. AB - This study was designed to identify the role of a recently identified Ca(2+)/calmodulin-dependent protein kinase (CaMK)-like kinase (CaMKLK) in neuronal apoptosis. For this purpose, we studied proteolytic cleavage of CaMKLK by caspases in vitro and in neuronal NG108 cells. In addition, we have investigated the effect of overexpression of wild type and mutant CaMKLK proteins on staurosporine- and serum deprivation-induced apoptosis of NG108 cells. We found that CaMKLK is a substrate for caspase-3 and -8, both in vitro and in NG108 cells during staurosporine- and serum withdrawal-induced apoptosis. Substitution of an aspartic acid residue at position 62 in an asparagine residue within a putative caspase cleavage site completely blocked cleavage of CaMKLK, strongly indicating that (59)DEND(62) is the caspase recognition site. Overexpression of an Asp(62) --> Asn CaMKLK mutant protected NG108 cells from staurosporine-induced apoptosis to a similar extent as Bcl-x(L). In contrast, overexpression of wild type CaMKLK did not lead to protection. Moreover, microinjection of Asp(62) --> Asn CaMKLK protected NG108 cells from serum deprivation-induced apoptosis, while overexpression of a caspase-generated noncatalytic N-terminal CaMKLK fragment exacerbated apoptosis. Together, our data suggest that cleavage of CaMKLK and generation of the noncatalytic N-terminal domain of CaMKLK facilitate neuronal apoptosis. PMID- 11479290 TI - Halotolerant cyanobacterium Aphanothece halophytica contains an Na(+)/H(+) antiporter, homologous to eukaryotic ones, with novel ion specificity affected by C-terminal tail. AB - Recently, a cyanobacterium Synechocystis sp. PCC 6803 has been shown to contain an Na(+)/H(+) antiporter gene homologous to plants (SOS1 and AtNHX1 from Arabidopsis) and mammalians (NHEs from human) but not to Escherichia coli (nhaA and nhaB). Here, we examined whether a halotolerant cyanobacterium Aphanothece halophytica has homologous genes. It turned out that A. halophytica contains an Na(+)/H(+) antiporter homologous to plants, mammalians, and some bacteria (nhaP from Pseudomonas and synnhaP from Synechocystis) but with novel ion specificity. Its gene product, ApNhaP (Na(+)/H(+) antiporter from Aphanothece halophytica), exhibited the Na(+)/H(+) antiporter activity over a wide pH range between 5 and 9 and complemented the Na(+)-sensitive phenotype of the antiporter-deficient E. coli mutant. The ApNhaP had virtually no activity for the Li(+)/H(+) antiporter but showed high Ca(2+)/H(+) antiporter activity at alkaline pH. The ApNhaP complemented the Ca(2+)-sensitive phenotype of the E. coli mutant but not the Li(+)-sensitive phenotype. The replacement of a long C-terminal tail of ApNhaP with that of Synechocystis altered the ion specificity of the antiporter. These results suggest that the ion specificity of an Na(+)/H(+) antiporter is partly determined by the structural properties of the C-terminal tail, which was well exemplified in the case of A. halophytica. PMID- 11479291 TI - Location and orientation of minK within the I(Ks) potassium channel complex. AB - The slowly activating cardiac potassium current (I(Ks)) is generated by a heteromultimeric potassium channel complex consisting of pore-forming (KvLQT1) and accessory (minK) subunits belonging to the KCNQ and KCNE gene families, respectively. Evidence indicating that minK residues line the I(Ks) pore originates from the observation that two minK cysteine mutants (G55C and F54C) render I(Ks) Cd2+-sensitive. We have identified a single cysteine residue in the KvLQT1 S6 segment (Cys-331) that contributes to Cd2+ coordination in conjunction with cysteine residues engineered into the minK transmembrane domain. This observation indicates that minK resides in close proximity to S6 in the I(Ks) channel complex. On the basis of homology modeling that compares the KvLQT1 S6 segment with the structure of the bacterial potassium channel KcsA, we predict that the sulfhydryl side chain of Cys-331 projects away from the central axis of the KvLQT1 pore and suggest that minK resides outside of the permeation pathway. A preliminary model illustrating the orientation of minK with S6 was validated by successful prediction of a novel Cd2+ binding site created within the I(Ks) channel complex by engineering additional cysteine residues into both subunits. Our results indicate the location and orientation of minK within the I(Ks) channel complex and further suggest that Cd2+ exerts its effect on I(Ks) through an allosteric mechanism rather than direct pore blockade. PMID- 11479292 TI - Analysis of the CD151-alpha3beta1 integrin and CD151-tetraspanin interactions by mutagenesis. AB - Transmembrane proteins of the tetraspanin superfamily are associated with various integrins and modulate their function. We performed mutagenesis analysis to establish structural requirements for the interaction of CD151 with the alpha3beta1 integrin and with other tetraspanins. Using a panel of CD151/CD9 chimeras and CD151 deletion mutants we show that the minimal region, which confers stable (e.g. Triton X-100-resistant) association of the tetraspanin with alpha3beta1, maps within the large extracellular loop (LECL) of CD151 (the amino acid sequence between residues Leu(149) and Glu(213)). Furthermore, the substitution of 11 amino acids (residues 195-205) from this region for a corresponding sequence from CD9 LECL or point mutations of cysteines in the conserved CCG and PXXCC motifs abolish the interaction. The removal of the LECL CD151 does not affect the association of the protein with other tetraspanins (e.g. CD9, CD81, CD63, and wild-type CD151). On the other hand, the mutation of the CCG motif selectively prevents the homotypic CD151-CD151 interaction but does not influence the association of the mutagenized CD151 with other tetraspanins. These results demonstrate the differences in structural requirements for the heterotypic and homotypic tetraspanin-tetraspanin interactions. Various deletions involving the small extracellular loop and the first three transmembrane domains prevent surface expression of the CD151 mutants but do not affect the CD151 alpha3beta1 interaction. The CD151 deletion mutants are accumulated in the endoplasmic reticulum and redirected to the lysosomes. The assembly of the CD151 alpha3beta1 complex occurs early during the integrin biosynthesis and precedes the interaction of CD151 with other tetraspanins. Collectively, these data show that the incorporation of CD151 into the "tetraspanin web" can be controlled at various levels by different regions of the protein. PMID- 11479293 TI - The role of CD47 in neutrophil transmigration. Increased rate of migration correlates with increased cell surface expression of CD47. AB - CD47, a cell surface glycoprotein, plays an important role in modulating neutrophil (PMN) migration across endothelial and epithelial monolayers. Here we show that anti-CD47 monoclonal antibodies (mAbs) delay PMN migration across collagen-coated filters or T84 epithelial monolayers toward the chemoattractant formylmethionylleucylphenylalanine (fMLP). Despite delayed transmigration by anti CD47 mAbs, the numbers of PMN migrating across in either condition were the same as in the presence of control non-inhibitory mAbs. Cell surface labeling and immunoprecipitation demonstrated upregulation of CD47 to the PMN cell surface with kinetics similar to those of the transmigration response. Subcellular fractionation studies revealed redistribution of CD47 from intracellular compartments that co-sediment with secondary granules to plasma membrane containing fractions after fMLP stimulation. Experiments performed to investigate potential signaling pathways revealed that inhibition of tyrosine phosphorylation with genistein reversed the anti-CD47-mediated PMN migration delay, whereas inhibition of phosphatidylinositol 3-kinase only partially reversed anti-CD47 effects that correlated with a rapid increase in PMN cell surface CD47. Analysis of the contribution of epithelial-expressed CD47 to PMN transmigration revealed that PMN migration across CD47-deficient epithelial monolayers (CaCO2) was significantly increased after stable transfection with CD47. These results suggest that cell surface CD47 and downstream tyrosine phosphorylation signaling events regulate, in part, the rate of PMN migration during the inflammatory response. PMID- 11479294 TI - A cholera toxin B-subunit variant that binds ganglioside G(M1) but fails to induce toxicity. AB - Entry of cholera toxin (CT) into target epithelial cells and the induction of toxicity depend on CT binding to the lipid-based receptor ganglioside G(M1) and association with detergent-insoluble membrane microdomains, a function of the toxin's B-subunit. The B-subunits of CT and related Escherichia coli toxins exhibit a highly conserved exposed peptide loop (Glu(51)-Ile(58)) that faces the cell membrane upon B-subunit binding to G(M1). Mutation of His(57) to Ala in this loop resulted in a toxin (CT-H57A) that bound G(M1) with high apparent affinity, but failed to induce toxicity. CT-H57A bound to only a fraction of the cell surface receptors available to wild-type CT. The bulk of cell-surface receptors inaccessible to CT-H57A localized to detergent-insoluble apical membrane microdomains (lipid rafts). Compared with wild-type toxin, CT-H57A exhibited slightly lower apparent binding affinity for and less stable binding to G(M1) in vitro. Rather than being transported into the Golgi apparatus, a process required for toxicity, most of CT-H57A was rapidly released from intact cells at physiologic temperatures or degraded following its internalization. These data indicate that CT action depends on the stable formation of the CT B-subunit.G(M1) complex and provide evidence that G(M1) functions as a necessary sorting motif for the retrograde trafficking of toxin into the secretory pathway of target epithelial cells. PMID- 11479295 TI - Cytokine-induced activation of nuclear factor-kappa B is inhibited by hydrogen peroxide through oxidative inactivation of IkappaB kinase. AB - Rapid activation of the IkappaB kinase (IKK) complex is considered an obligatory step in the activation of nuclear factor-kappaB (NF-kappaB) in response to diverse stimuli. Since oxidants have been implicated in the regulation of NF kappaB, the focus of the present study was the activation of IKK by tumor necrosis factor alpha (TNFalpha) in the presence or absence of hydrogen peroxide (H(2)O(2)). Exposure of mouse alveolar epithelial cells to H(2)O(2) was not sufficient to activate IKK, degrade IkappaBalpha, or activate NF-kappaB. In contrast, TNFalpha induced IKK activity rapidly and transiently resulting in IkappaBalpha degradation and NF-kappaB activation. Importantly, in the presence of H(2)O(2), the ability of TNFalpha to induce IKK activity was markedly decreased and resulted in prevention of IkappaBalpha degradation and NF-kappaB activation. Neither tyrosine kinases nor phosphatidylinositol 3-kinases, known regulators of NF-kappaB by oxidants, were involved in IKK inhibition by H(2)O(2). Direct addition of H(2)O(2) to the immunoprecipitated IKK complex inhibited enzyme activity. Inhibition of IKK activity by H(2)O(2) was associated with direct oxidation of cysteine residues present in the IKK complex and occurred only in enzymatically active IKK. In contrast to previously published observations, our findings demonstrate that the oxidant H(2)O(2) reduces NF kappaB activation by inhibiting activated IKK activity. PMID- 11479296 TI - Functional analysis of the four DNA binding domains of replication protein A. The role of RPA2 in ssDNA binding. AB - Replication Protein A (RPA), the heterotrimeric single-stranded DNA (ssDNA) binding protein of eukaryotes, contains four ssDNA binding domains (DBDs) within its two largest subunits, RPA1 and RPA2. We analyzed the contribution of the four DBDs to ssDNA binding affinity by assaying recombinant yeast RPA in which a single DBD (A, B, C, or D) was inactive. Inactivation was accomplished by mutating the two conserved aromatic stacking residues present in each DBD. Mutation of domain A had the most severe effect and eliminated binding to a short substrate such as (dT)12. RPA containing mutations in DBDs B and C bound to substrates (dT)12, 17, and 23 but with reduced affinity compared with wild type RPA. Mutation of DBD-D had little or no effect on the binding of RPA to these substrates. However, mutations in domain D did affect the binding to oligonucleotides larger than 23 nucleotides (nt). Protein-DNA cross-linking indicated that DBD-A (in RPA1) is essential for RPA1 to interact efficiently with substrates of 12 nt or less and that DBD-D (RPA2) interacts efficiently with oligonucleotides of 27 nt or larger. The data support a sequential model of binding in which DBD-A is responsible for the initial interaction with ssDNA, that domains A, B, and C (RPA1) contact 12-23 nt of ssDNA, and that DBD-D (RPA2) is needed for RPA to interact with substrates that are 23-27 nt in length. PMID- 11479297 TI - Acetylation of steroidogenic factor 1 protein regulates its transcriptional activity and recruits the coactivator GCN5. AB - Steroidogenic factor-1 (SF-1) is an orphan nuclear receptor that plays an essential role in the development of the hypothalamic-pituitary-gonadal axis in both sexes. SF-1 belongs to the hormone nuclear receptor superfamily and possesses an N-terminal DNA binding domain and a C-terminal ligand binding domain. Activation function domain 2 is located C-terminal of the ligand binding domain of SF-1 and is important for the transactivation of target genes. Coactivators with histone acetyltransferase activity such as cAMP response element-binding protein-binding protein and steroid receptor coactivator 1 interact and increase SF-1-mediated transcriptional activity. In this study we demonstrate that SF-1 is acetylated in vivo. Histone acetyltransferase GCN5 acetylates SF-1 in vitro. Moreover, we found that SF-1 recruited a novel coactivator GCN5, which can be a newly identified coactivator for SF-1. Acetylation of SF-1 stimulates its transcriptional activity. Inhibition of deacetylation by trichostatin A, a histone deacetylase inhibitor, increased SF-1 mediated transactivation and stabilized and induced the nuclear export of the SF 1 protein. PMID- 11479298 TI - The classical progesterone receptor associates with p42 MAPK and is involved in phosphatidylinositol 3-kinase signaling in Xenopus oocytes. AB - The induction of Xenopus laevis oocyte maturation by progesterone is a striking example of a steroid hormone-mediated event that does not require transcription. Here we have investigated the role of the classical progesterone receptor in this nongenomic signaling. The Xenopus progesterone receptor (XPR) was predominantly cytoplasmic; however, a significant fraction ( approximately 5%) of one form of the receptor (p82 XPR) was associated with the plasma membrane-containing P 10,000 fraction, compatible with the observation that membrane-impermeant derivatives of progesterone can induce maturation. XPR co-precipitated with active phosphatidylinositol 3-kinase. The phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin delayed progesterone-induced maturation and completely blocked the insulin-dependent maturation, indicating that the association of XPR with PI3-K could be functionally important. We also examined whether the nongenomic signaling properties of XPR can account for the ability of glucocorticoids and the progesterone antagonist RU486 to induce maturation. We found that none of these steroids cause XPR to become associated with active PI3 K; thus, association of XPR with active PI3-K is progesterone-specific. Finally, we showed that p42 mitogen-activated protein kinase (MAPK) associates with XPR after progesterone-induced germinal vesicle breakdown and that active recombinant MAPK is able to phosphorylate p110 XPR in vitro. These findings demonstrate that the classical progesterone receptor is involved in progesterone-induced nongenomic signaling in Xenopus oocytes and provide evidence that p42 MAPK and PI3-K activity are directly associated with the classical progesterone receptor. PMID- 11479299 TI - Hormone-mediated dephosphorylation of specific histone H1 isoforms. AB - We have previously shown a connection between histone H1 phosphorylation and the transcriptional competence of the hormone inducible mouse mammary tumor virus (MMTV) promoter. Prolonged exposure of mouse cells to dexamethasone concurrently dephosphorylated histone H1 and rendered the MMTV promoter refractory to hormonal stimulation and, therefore, transcriptionally unresponsive. Using electrospray mass spectrometry, we demonstrate here that prolonged dexamethasone treatment differentially effects a subset of the six somatic H1 isoforms in mouse cells. H1 isoforms H1.0, H1.1, and H1.2 are non-responsive to hormone whereas prolonged dexamethasone treatment effectively dephosphorylated the H1.3, H1.4, and H1.5 isoforms. The protein kinase inhibitor staurosporine, shown to dephosphorylate histone H1 and down-regulate MMTV in cultured cells, appears only to completely dephosphorylate the H1.3 isoform. These results suggest that dephosphorylation of specific histone H1 isoforms may contribute to the previously observed decrease in transcriptional competence of the MMTV promoter through the modulation of chromatin structure. In a broader sense, this work advances the hypothesis that post-translational modifications of individual histone H1 isoforms directly influence the transcriptional activation/repression of specific genes. PMID- 11479300 TI - Engineering regulable Escherichia coli beta-galactosidases as biosensors for anti HIV antibody detection in human sera. AB - The activity of engineered, peptide-displaying enzymes is modulated by binding to specific anti-peptide antibodies. This new concept of a quantitative antibody detection system allows test kits to be set up for fast diagnosis of infectious diseases. To develop a quick and homogeneous assay for the detection of human immunodeficiency virus (HIV) infection, we have explored two acceptor sites of the bacterial Escherichia coli beta-galactosidase for the accommodation of HIV antigenic peptides. Two overlapping epitopes (namely P1 and P2) from the gp41 envelope glycoprotein, contained in different sized peptides, were inserted in the vicinity of the enzyme active site to generate a set of hybrid, enzymatically active beta-galactosidases. Regulable enzymes of different responsiveness to monoclonal antibody binding were generated with both acceptor sites tested. These biosensors were also sensitive to immune sera from HIV-infected patients. Modeling data provide insight into the structural modifications in the vicinity of the active site induced by peptide insertion that strongly affect the responsiveness of the engineered proteins through different parameters of their catalytic properties. PMID- 11479301 TI - A model of reversible inhibitors in the gastric H+/K+-ATPase binding site determined by rotational echo double resonance NMR. AB - Several close analogues of the noncovalent H(+)/K(+)-ATPase inhibitor SCH28080 (2 methyl-3-cyanomethyl-8-(phenylmethoxy)imidazo[1,2-a]pyridine) have been screened for activity and examined in the pharmacological site of action by solid-state NMR spectroscopy. TMPIP, the 1,2,3-trimethyl analogue of SCH28080, and variants of TMPIP containing fluorine in the phenylmethoxy ring exhibited IC(50) values for porcine H(+)/K(+)-ATPase inhibition falling in the sub-10 microm range. Deuterium NMR spectra of a (2)H-labeled inhibitor titrated into H(+)/K(+)-ATPase membranes revealed that 80-100% of inhibitor was bound to the protein, and K(+) competition (2)H NMR experiments confirmed that the inhibitor lay within the active site. The active binding conformation of the pentafluorophenylmethoxy analogue of TMPIP was determined from (13)C-(19)F dipolar coupling measurements using the cross-polarization magic angle spinning NMR method, REDOR. It was found that the inhibitor adopts an energetically favorable extended conformation falling between fully planar and partially bowed extremes. These findings allowed a model to be proposed for the binding of this inhibitor to H(+)/K(+)-ATPase based on the results of independent site-directed mutagenesis studies. In the model, the partially bowed inhibitor interacts with Phe(126) close to the N terminal membrane spanning helix M1 and residues in the extracellular loop bridging membrane helices M5 and M6 and is flanked by residues in M4. PMID- 11479302 TI - Critical roles of TRAF2 and TRAF5 in tumor necrosis factor-induced NF-kappa B activation and protection from cell death. AB - Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) were identified as signal transducers for the TNF receptor superfamily. However, the exact roles of TRAF2 and TRAF5 in TNF-induced NF-kappaB activation still remain controversial. To address this issue, we generated TRAF2 and TRAF5 double knockout (DKO) mice. TNF- but not interleukin-1-induced nuclear translocation of NF-kappaB was severely impaired in murine embryonic fibroblasts (MEFs) derived from DKO mice. Moreover, DKO MEFs were more susceptible to TNF-induced cytotoxicity than TRAF2 knockout MEFs. Collectively, these results indicate that both TRAF2 and TRAF5 are involved in TNF-induced NF-kappaB activation and protection from cell death. PMID- 11479303 TI - Coupled, but not uncoupled, fluxes in a neuronal glutamate transporter can be activated by lithium ions. AB - In the central nervous system a family of related (Na(+)-K(+))-coupled glutamate transporters remove the transmitter from the cleft and prevent its neurotoxic actions. In addition to this coupled uptake, these transporters also mediate a sodium- and glutamate-dependent uncoupled anion conductance. Most models assume that the initial steps for both processes are the same, leading to the anticipation that both may exhibit a similar requirement for cations. In this study we have tested this idea in the neuronal glutamate transporter EAAC-1. We report that in this transporter lithium can replace sodium in the coupled uptake. Strikingly, the glutamate-dependent gating of the uncoupled conductance mediated by EAAC-1 has a strict requirement for sodium; lithium cannot substitute for it. Moreover, we describe two mutants, T370S and G410S, in which the cation selectivity of the two processes is affected differently. In both mutants sodium, but not lithium, can support coupled transport. On the other hand, the sodium selectivity of the gated anion conductance in oocytes expressing the mutant transporters is not affected. Our observations indicate that although both the coupled and the uncoupled fluxes are sodium-dependent, the conformation gating the anion conductance is different from that during substrate translocation. PMID- 11479304 TI - Are 5-hydroxytryptamine-preloaded beta-cells an appropriate physiologic model system for establishing that insulin stimulates insulin secretion? AB - The release and oxidation of 5-hydroxytryptamine from 5-hydroxytryptamine preloaded beta-cells has been used as a surrogate marker for insulin secretion. Findings made using this methodology have been used to support the concept that insulin stimulates its own release. In the present studies, the effects of 5 hydroxytryptamine on stimulated insulin secretion from isolated perifused rat islets was determined. When added together with stimulatory glucose, 5 hydroxytryptamine (0.5 mm) significantly reduced both phases of 8 mm glucose induced secretion and reduced the first phase of 15 mm glucose-induced release by 60% without any effect on sustained insulin release rates. Preloading of beta cells with 0.5 mm 5-hydroxytryptamine for 3 h resulted in a more severe impairment of 15 mm glucose-induced secretion. First and second phase release rates were reduced by 70 and 55%, respectively. In addition, this pretreatment protocol also abolished 200 microm tolbutamide-induced insulin secretion from perifused islets. These findings confirm that 5-hydroxytryptamine is a powerful inhibitor of stimulated insulin secretion. The responses of 5-hydroxytryptamine preloaded beta-cells may not accurately reflect the biochemical events occurring during the physiologic regulation of insulin secretion. The suggestion that insulin stimulates its own secretion based exclusively on amperometric measurements should be reconsidered. PMID- 11479305 TI - Calcium-G protein interactions in the regulation of macrophage secretion. AB - The interplay between activated G proteins and intracellular calcium ([Ca(2+)](i)) in the regulation of secretion was studied in the macrophage, coupling membrane capacitance with calcium-sensitive microfluorimetry. Intracellular elevation of either the nonhydrolyzable analogue of GTP, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), or [Ca(2+)](i) enhanced the amplitude and shortened the time course of stimulus-induced secretion in a dose-dependent manner. Both the ionophore- and the stimulus-induced secretory response were abolished in the presence of guanosine-5'-O-(2-thiodiphosphate) (GDP beta S). The K(d) of Ca(2+)-driven secretion was independent of GTP gamma S concentration, whereas the K(d) of the GTP gamma S-driven response decreased from 63 to 31 microM in the presence of saturating concentrations of [Ca(2+)](i). The time course of stimulus-induced secretion was dependent upon the concentration of [Ca(2+)](i). The time course of GTP gamma S-driven secretion was concentration independent at high levels of [Ca(2+)](i), suggesting that a calcium-dependent translocation/binding step was rate-limiting. Our data strongly support a model in which [Ca(2+)](i) and activated G proteins act independently of one another in the sequential regulation of macrophage secretion. [Ca(2+)](i) appears to play a role in the recruitment and priming of vesicles from reserve intracellular pools at a step that is upstream of G protein activation. While activated, G proteins appear to play a key role in fusion of docked vesicles. Thus, secretion can result either from activating more G proteins or from elevating [Ca(2+)](i) at basal levels of G protein activation. PMID- 11479306 TI - Cyclic AMP inhibits extracellular signal-regulated kinase and phosphatidylinositol 3-kinase/Akt pathways by inhibiting Rap1. AB - Cyclic AMP inhibited both ERK and Akt activities in rat C6 glioma cells. A constitutively active form of phosphatidylinositol 3-kinase (PI3K) prevented cAMP from inhibiting Akt, suggesting that the inactivation of Akt by cAMP is a consequence of PI3K inhibition. Neither protein kinase A nor Epac (Exchange protein directly activated by cAMP), two known direct effectors of cAMP, mediated the cAMP-induced inhibition of ERK and Akt phosphorylation. Cyclic AMP inhibited Rap1 activation in C6 cells. Moreover, inhibition of Rap1 by a Rap1 GTPase activating protein-1 also resulted in a decrease in ERK and Akt phosphorylation, which was not further decreased by cAMP, suggesting that cAMP inhibits ERK and Akt by inhibiting Rap1. The role of Rap1 in ERK and Akt activity was further demonstrated by our observation that an active form of Epac, which activated Rap1 in the absence of cAMP, increased ERK and Akt phosphorylation. Inhibition of ERK and/or PI3K pathways mediated the inhibitory effects of cAMP on insulin-like growth factor-I (IGF-I) and IGF-binding protein-3 gene expression. Moreover, cAMP, as well as ERK and PI3K inhibitors produced equivalent stimulation and inhibition, respectively, of p27(Kip1) and cyclin D2 protein levels, potentially explaining the observation that cAMP prevented C6 cells from entering S phase. PMID- 11479307 TI - Low density lipoprotein receptor-related protein is required for macrophage mediated oxidation of low density lipoprotein by 12/15-lipoxygenase. AB - The oxidative modification of low density lipoprotein (LDL) has been implicated in the early stage of atherosclerosis through multiple potential pathways, and 12/15-lipoxygenase is suggested to be involved in this oxidation process. We demonstrated previously that the 12/15-lipoxygenase overexpressed in mouse macrophage-like J774A.1 cells was required for the cell-mediated LDL oxidation. However, the mechanism of the oxidation of extracellular LDL by the intracellular 12/15-lipoxygenase has not yet been elucidated. In the present study, we found that not only the LDL receptor but also LDL receptor-related protein (LRP), both of which are cell surface native LDL-binding receptors, were down-regulated by the preincubation of the cells with cholesterol or LDL and up-regulated by lipoprotein-deficient serum. Moreover, 12/15-lipoxygenase-expressing cell mediated LDL oxidation was decreased by the preincubation of the cells with LDL or cholesterol and increased by the preincubation with lipoprotein-deficient serum. Heparin-binding protein 44, an antagonist of the LDL receptor family, also suppressed the cell-mediated LDL oxidation in a dose-dependent manner. The cell mediated LDL oxidation was dose-dependently blocked by an anti-LRP antibody but not by an anti-LDL receptor antibody. Furthermore, antisense oligodeoxyribonucleotides against LRP reduced the cell-mediated LDL oxidation under the conditions in which the expression of LRP was decreased. The results taken together indicate that LRP was involved essentially for the cell-mediated LDL oxidation by 12/15-lipoxygenase expressed in J774A.1 cells, suggesting an important pathophysiological role of this receptor-enzyme system as the initial trigger of the progression of atherosclerosis. PMID- 11479308 TI - Cellular stress regulates the nucleocytoplasmic distribution of the protein tyrosine phosphatase TCPTP. AB - Specific cellular stresses, including hyperosmotic stress, caused a dramatic but reversible cytoplasmic accumulation of the otherwise nuclear 45-kDa variant of the protein-tyrosine phosphatase TCPTP (TC45). In the cytoplasm, TC45 dephosphorylated the epidermal growth factor receptor and down-regulated the hyperosmotic stress-induced activation of the c-Jun N-terminal kinase. The hyperosmotic stress-induced nuclear exit of TC45 was not inhibited by leptomycin B, indicating that TC45 nuclear exit was independent of the exportin CRM-1. Moreover, hyperosmotic stress did not induce the cytoplasmic accumulation of a green fluorescent protein-TC45 fusion protein that was too large to diffuse across the nuclear pore. Our results indicate that TC45 nuclear exit may occur by passive diffusion and that cellular stress may induce the cytoplasmic accumulation of TC45 by inhibiting nuclear import. Neither p42(Erk2) nor the stress-activated c-Jun N-terminal kinase or p38 mediated the stress-induced redistribution of TC45. We found that only those stresses that stimulated the metabolic stress-sensing enzyme AMP-activated protein kinase (AMPK) induced the redistribution of TC45. In addition, specific pharmacological activation of the AMPK was sufficient to cause the accumulation of TC45 in the cytoplasm. Our studies indicate that specific stress-activated signaling pathways that involve the AMPK can alter the nucleocytoplasmic distribution of TC45 and thus regulate TC45 function in vivo. PMID- 11479309 TI - Cyanobacteria perceive nitrogen status by sensing intracellular 2-oxoglutarate levels. AB - The regulatory circuits that control nitrogen metabolism are relatively well known in several bacterial model groups. However, much less is understood about how the nitrogen status of the cell is perceived in vivo. In cyanobacteria, the transcription factor NtcA is required for regulation (activation or repression) of an extensive number of genes involved in nitrogen metabolism. In contrast, how NtcA activity is regulated is largely unknown. Assimilation of ammonium by most microorganisms occurs through the sequential action of two enzymes: glutamine synthetase (GS) and glutamate synthase. Interestingly, regulation of the expression of NtcA-dependent genes in the cyanobacterium Synechocystis sp. PCC 6803 is altered in mutants with modified levels of GS activity. Two types of mutants were analyzed: glnA null mutants that lack GS type I and gif mutants unable to inactivate GS in the presence of ammonium. Changes in the intracellular pools of 19 different amino acids and the keto acid 2-oxoglutarate were recorded in wild-type and mutant strains under different nitrogen conditions. Our data strongly indicate that the nitrogen status in cyanobacteria is perceived as changes in the intracellular 2-oxoglutarate pool. PMID- 11479310 TI - Regulation of the properties of the heme-NO complexes in nitric-oxide synthase by hydrogen bonding to the proximal cysteine. AB - Nitric-oxide synthase (NOS) catalyzes the formation of NO and citrulline from l arginine and oxygen. However, the NO so formed has been found to auto-inhibit the enzymatic activity significantly. We hypothesized that the NO reactivity is in part controlled by hydrogen bonding between the conserved tryptophan residue (position 409 in the neuronal isoform of NOS (nNOS)) and the cysteine residue that forms the proximal bond to the heme. By using resonance Raman spectroscopy and NO as a probe of the heme environment, we show that in the W409F and W409Y mutants of the oxygenase domain of the neuronal enzyme (nNOSox), the Fe-NO bond in the Fe3+NO complex is weaker than in the wild type enzyme, consistent with the loss of a hydrogen bond on the sulfur atom of the proximal cysteine residue. The weaker Fe-NO bond in the W409F and W409Y mutants might result in a faster rate of NO dissociation from the ferric heme in the Trp-409 mutants as compared with the wild type enzyme, which could contribute to the lower accumulation of the inhibitory NO-bound complexes observed during catalysis with the Trp-409 mutants (Adak, S., Crooks, C., Wang, Q., Crane, B. R., Tainer, J. A., Getzoff, E. D., and Stuehr, D. J. (1999) J. Biol. Chem. 274, 26907-26911). The optical and resonance Raman spectra of the Fe2+NO complexes of the Trp-409 mutants differ from those of the wild type enzyme and indicate that a significant population of a five coordinate Fe2+NO complex is present. These data show that the hydrogen bond provided by the Trp-409 residue is necessary to maintain the thiolate coordination when NO binds to the ferrous heme. Taken together our results indicate that the heme environment on the proximal side of nNOS is critical for the formation of a stable iron-cysteine bond and for the control of the electronic properties of heme-NO complexes. PMID- 11479311 TI - Major histocompatibility complex class I-presented antigenic peptides are degraded in cytosolic extracts primarily by thimet oligopeptidase. AB - Nearly all peptides generated by proteasomes during protein degradation are digested rapidly to amino acids, but a few proteasomal products escape this fate and are presented to the immune system on cell surface major histocompatibility complex class I molecules. To test whether these antigenic peptides may be inherently resistant to cytosolic peptidases, six different antigenic peptides were incubated with HeLa cell extracts. All six were degraded rapidly by a process involving o-phenanthroline-sensitive metallopeptidases. One antigenic peptide, FAPGNYPAL, was rapidly destroyed in the extracts by a bestatin-sensitive exopeptidase, apparently by the puromycin-sensitive aminopeptidase. The disappearance of the other five was reduced 30-90% by a specific inhibitor of the cytosolic endopeptidase, thimet oligopeptidase (TOP) (EC ), whose physiological function(s) have been unclear and controversial. All these peptides were sensitive to pure recombinant TOP. Furthermore, upon fractionation of the extracts, the major peptidase peak that degraded the ovalbumin-derived epitope, SIINFEKL, co-purified with TOP. In the extracts, TOP also catalyzed rapid degradation of N-extended variants of SIINFEKL and of other antigenic peptides, which in vivo can serve as precursors of these major histocompatibility complex presented epitopes. This enzyme (unlike cell proteins that promote production of antigenic peptides) is not regulated by interferon-gamma. TOP seems to be primarily responsible for the rapid breakdown of antigenic peptides in cytosolic extracts, and our related studies (A. X. Y. Mo, K. Lemerise, W. Zeng, Y. Shen, C. R. Abraham, A. L. Goldberg, and K. L. Rock, submitted for publication) indicate that TOP by destroying such peptides limits antigen presentation in vivo. PMID- 11479312 TI - Plasmodium falciparum possesses a classical glutaredoxin and a second, glutaredoxin-like protein with a PICOT homology domain. AB - The genes coding for two different proteins with homologies to glutaredoxins have been identified in the genome of the malarial parasite Plasmodium falciparum. Both genes were amplified from a gametocytic cDNA and overexpressed in Escherichia coli. The smaller protein (named PfGrx-1) with 12.4 kDa in size exhibits the typical glutaredoxin active site motif "CPYC," shows glutathione dependent glutaredoxin activity in the beta-hydroxyethyl disulfide (HEDS) assay, and reduces Trypanosoma brucei ribonucleotide reductase. Glutathione:HEDS transhydrogenase activity (approximately 60 milliunits/mg of protein) was clearly detectable in trophozoite extracts from eight different P. falciparum strains and did not differ between chloroquine-resistant and -sensitive parasites. Five different antimalarial drugs at 100 microm did not significantly influence isolated PfGrx-1 activity. In contrast, the second protein (deduced mass 19.9 kDa) with homology to glutaredoxins (31% identity to Schizosaccharomyces pombe in a 140-amino acid overlap) was not active in the HEDS assay; however, its general dithiol reducing activity was demonstrated in the insulin assay in the presence of dithiothreitol. Interestingly, the sequence contains a PICOT (for protein kinase C-interacting cousin of thioredoxin) homology domain, which might suggest regulatory functions of the protein. We named this protein PfGLP-1, for P. falciparum 1-Cys-glutaredoxin-like protein-1. In contrast to glutaredoxins, PfGLP 1 could not be reduced by glutathione. This is the first report on glutaredoxin like proteins in the family of Plasmodia. PMID- 11479313 TI - alpha 2,3-Sialylation of terminal GalNAc beta 1-3Gal determinants by ST3Gal II reveals the multifunctionality of the enzyme. The resulting Neu5Ac alpha 2 3GalNAc linkage is resistant to sialidases from Newcastle disease virus and Streptococcus pneumoniae. AB - Enzymatic alpha 2,3-sialylation of GalNAc has not been described previously, although some glycoconjugates containing alpha 2,3-sialylated GalNAc residues have been reported. In the present experiments, recombinant soluble alpha 2,3 sialyltransferase ST3Gal II efficiently sialylated the X(2) pentasaccharide GalNAc beta 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc, globo-N-tetraose GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc, and the disaccharide GalNAc beta 1 3Gal in vitro. The purified products were identified as Neu5Ac alpha 2-3GalNAc beta 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc, Neu5Ac alpha 2-3GalNAc beta 1 3Gal alpha 1-4Gal beta 1-4Glc, and Neu5Ac alpha 2-3GalNAc beta 1-3Gal, respectively, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, enzymatic degradations, and one- and two-dimensional NMR spectroscopy. In particular, the presence of the Neu5Ac alpha 2-3GalNAc linkage was firmly established in all three products by a long range correlation between Neu5Ac C2 and GalNAc H3 in heteronuclear multiple bond correlation spectra. Collectively, the data describe the first successful sialyltransfer reactions to the 3-position of GalNAc in any acceptor. Previously, ST3Gal II has been shown to transfer to the Gal beta 1-3GalNAc determinant. Consequently, the present data show that the enzyme is multifunctional, and could be renamed ST3Gal(NAc) II. In contrast to ST3Gal II, ST3Gal III did not transfer to the X(2) pentasaccharide. The Neu5Ac alpha 2-3GalNAc linkage of sialyl X(2) was cleaved by sialidases from Arthrobacter ureafaciens and Clostridium perfringens, but resisted the action of sialidases from Newcastle disease virus and Streptococcus pneumoniae. Therefore, the latter two enzymes cannot be used to differentiate between Neu5Ac alpha 2 3GalNAc and Neu5Ac alpha 2-6GalNAc linkages, as has been assumed previously. PMID- 11479314 TI - Folding and aggregation are selectively influenced by the conformational preferences of the alpha-helices of muscle acylphosphatase. AB - The native state of human muscle acylphosphatase (AcP) presents two alpha helices. In this study we have investigated folding and aggregation of a number of protein variants having mutations aimed at changing the propensity of these helical regions. Equilibrium and kinetic measurements of folding indicate that only helix-2, spanning residues 55-67, is largely stabilized in the transition state for folding therefore playing a relevant role in this process. On the contrary, the aggregation rate appears to vary only for the variants in which the propensity of the region corresponding to helix-1, spanning residues 22-32, is changed. Mutations that stabilize the first helix slow down the aggregation process while those that destabilize it increase the aggregation rate. AcP variants with the first helix destabilized aggregate with rates increased to different extents depending on whether the introduced mutations also alter the propensity to form beta-sheet structure. The fact that the first alpha-helix is important for aggregation and the second helix is important for folding indicates that these processes are highly specific. This partitioning does not reflect the difference in intrinsic alpha-helical propensities of the two helices, because helix-1 is the one presenting the highest propensity. Both processes of folding and aggregation do not therefore initiate from regions that have simply secondary structure propensities favorable for such processes. The identification of the regions involved in aggregation and the understanding of the factors that promote such a process are of fundamental importance to elucidate the principles by which proteins have evolved and for successful protein design. PMID- 11479315 TI - PDZ interaction sites in integrin alpha subunits. T14853, TIP/GIPC binds to a type I recognition sequence in alpha 6A/alpha 5 and a novel sequence in alpha 6B. AB - We used published peptide library data to identify PDZ recognition sequences in integrin alpha subunit cytoplasmic domains and found that the alpha(6)A and alpha(5) subunits contain a type I PDZ binding site (TSDA*) (asterisk indicates the stop codon). The alpha(6)A cytoplasmic domain was used for screening a two hybrid library to find interacting proteins. The bulk of the captured cDNAs (60%) coded for TIP-2/GIPC, a cytoplasmic protein with one PDZ domain. The interaction of TIP-2/GIPC with different integrin subunits was tested in two-hybrid and in vitro binding assays. Surprisingly, TIP-2/GIPC bound strongly to the C terminus of both alpha(6)A and alpha(6)B, although the alpha(6)B sequence (ESYS*) is not suggestive of a PDZ binding site because of its polar C-terminal residue. For high affinity interaction with TIP-2/GIPC, at least one of the residues at positions -1 and -3 must be negatively charged. An aliphatic residue at position 0 increases the affinity of but is not required for this interaction. The alpha(5) integrin subunit also bound to TIP-2/GIPC. The alpha(6) integrin and TIP 2/GIPC co-localize in retraction fibers in carcinoma cells plated on laminin, a finding suggesting a functional interaction in vivo. Our results demonstrate that both splice variants of alpha(6) integrin contain a conserved PDZ binding site that enables interaction with TIP-2/GIPC. The binding site in alpha(6)B defines a new subclass of type I PDZ interaction site, characterized by a non-aliphatic residue at position 0. PMID- 11479316 TI - Appican, the proteoglycan form of the amyloid precursor protein, contains chondroitin sulfate E in the repeating disaccharide region and 4-O-sulfated galactose in the linkage region. AB - Chondroitin sulfate (CS)-D and CS-E, which are characterized by oversulfated disaccharide units, have been shown to regulate neuronal adhesion, cell migration, and neurite outgrowth. CS proteoglycans (CSPGs) consist of a core protein to which one or more CS chains are attached via a serine residue. Although several brain CSPGs, including mouse DSD-1-PG/phosphacan, have been found to contain the oversulfated D disaccharide motif, no brain CSPG has been reported to contain the oversulfated E motif. Here we analyzed the CS chain of appican, the CSPG form of the Alzheimer's amyloid precursor protein. Appican is expressed almost exclusively by astrocytes and has been reported to have brain- and astrocyte-specific functions including stimulation of both neural cell adhesion and neurite outgrowth. The present findings show that the CS chain of appican has a molecular mass of 25-50 kDa. This chain contains a significant fraction (14.3%) of the oversulfated E motif GlcUA beta 1-3GalNAc(4,6-O disulfate). The rest of the chain consists of GlcUA beta 1-3GalNAc(4-O-sulfate) (81.2%) and minor fractions of GlcUA beta 1-3GalNAc and GlcUA beta 1-3GalNAc(6-O sulfate). We also show that the CS chain of appican contains in its linkage region the 4-O-sulfated Gal structure. Thus, appican is the first example of a specific brain CSPG that contains the E disaccharide unit in its sugar backbone and the 4-O-sulfated Gal residue in its linkage region. The presence of the E unit is consistent with and may explain the neurotrophic activities of appican. PMID- 11479317 TI - Pleiotropic effects of post-translational modifications on the fate of viral glycopeptides as cytotoxic T cell epitopes. AB - The fate of viral glycopeptides as cytotoxic T lymphocyte (CTL) epitopes is unclear. We have dissected the mechanisms of antigen presentation and CTL recognition of the peptide GP392-400 (WLVTNGSYL) from the lymphocytic choriomeningitis virus (LCMV) and compared them with those of the previously reported GP92-101 antigen (CSANNSHHYI). Both GP392-400 and GP92-101 bear a glycosylation motif, are naturally N-glycosylated in the mature viral glycoproteins, bind to major histocompatibility complex H-2D(b) molecules, and are immunogenic. However, post-translational modifications differentially affected GP92-101 and GP392-400. Upon N-glycosylation or de-N-glycosylation, a marked decrease in major histocompatibility complex binding was observed for GP392-400 but not for GP92-101. Further, under its N-glycosylated or de-N glycosylated form, GP392-400 then lost its initial ability to generate a CTL response in mice, whereas GP92-101 was still immunogenic under the same conditions. The genetically encoded form of GP392-400, which on the basis of its immunogenicity could still be presented with H-2D(b) during the course of LCMV infection, does not in fact appear at the surface of LCMV-infected cells. Our results show that post-translational modifications of viral glycopeptides can have pleiotropic effects on their presentation to and recognition by CTL that contribute to either creation of neo-epitopes or destruction of potential epitopes. PMID- 11479318 TI - Identification of two topologically independent domains in RAG1 and their role in macromolecular interactions relevant to V(D)J recombination. AB - V(D)J recombination is instigated by the recombination-activating proteins RAG1 and RAG2, which catalyze site-specific DNA cleavage at the border of the recombination signal sequence (RSS). Although both proteins are required for activity, core RAG1 (the catalytically active region containing residues 384-1008 of 1040) alone displays binding specificity for the conserved heptamer and nonamer sequences of the RSS. The nonamer-binding region lies near the N terminus of core RAG1, whereas the heptamer-binding region has not been identified. Here, potential domains within core RAG1 were identified using limited proteolysis studies. An iterative procedure of DNA cloning, protein expression, and characterization revealed the presence of two topologically independent domains within core RAG1, referred to as the central domain (residues 528-760) and the C terminal domain (residues 761-980). The domains do not include the nonamer binding region but rather largely span the remaining relatively uncharacterized region of core RAG1. Characterization of macromolecular interactions revealed that the central domain bound to the RSS with specificity for the heptamer and contained the predominant binding site for RAG2. The C-terminal domain bound DNA cooperatively but did not show specificity for either conserved RSS element. This domain was also found to self-associate, implicating it as a dimerization domain within RAG1. PMID- 11479319 TI - Multiple human vault RNAs. Expression and association with the vault complex. AB - Human vaults are intracellular ribonucleoprotein particles believed to be involved in multidrug resistance. The complex consists of a major vault protein (MVP), two minor vault proteins (VPARP and TEP1), and several small untranslated RNA molecules. Three human vault RNA genes (HVG1-3) have been described, and a fourth was found in a homology search (HVG4). In the literature only the association of hvg1 with vaults was shown in vivo. However, in a yeast three hybrid screen the association of hvg1, hvg2, and hvg4 with TEP1 was demonstrated. In this study we investigated the expression and vault association of different vault RNAs in a variety of cell lines, including pairs of drug-sensitive and drug resistant cells. HVG1-3 are expressed in all cell lines examined, however, none of the cell lines expressed HVG4. This probably is a consequence of the absence of essential external polymerase III promoter elements. The bulk of the vault RNA associated with vaults was hvg1. Interestingly, an increased amount of hvg3 was bound to vaults isolated from multidrug-resistant cell lines. Our findings suggest that vaults bind the RNA molecules with different affinities in different situations. The ratio in which the vault RNAs are associated with vaults might be of functional importance. PMID- 11479320 TI - Conformational changes that occur during an RNA-editing adenosine deamination reaction. AB - ADARs are adenosine deaminases responsible for RNA-editing reactions that occur within duplex RNA. Currently little is known regarding the nature of the protein RNA interactions that lead to site-selective adenosine deamination. We previously reported that ADAR2 induced changes in 2-aminopurine fluorescence of a modified substrate, consistent with a base-flipping mechanism. Additional data have been obtained using full-length ADAR2 and a protein comprising only the RNA binding domain (RBD) of ADAR2. The increase in 2-aminopurine fluorescence is specific to the editing site and dependent on the presence of the catalytic domain. Hydroxyl radical footprinting demonstrates that the RBD protects a region of the RNA duplex around the editing site, suggesting a significant role for the RBD in identifying potential ADAR2 editing sites. Nucleotides near the editing site on the non-edited strand become hypersensitive to hydrolytic cleavage upon binding of ADAR2 RBD. Therefore, the RBD may assist base flipping by increasing the conformational flexibility of nucleotides in the duplex adjacent to its binding site. In addition, an increase in tryptophan fluorescence is observed when ADAR2 binds duplex RNA, suggesting a conformational change in the catalytic domain of the enzyme. Furthermore, acrylamide quenching experiments indicate that RNA binding creates heterogeneity in the solvent accessibility of ADAR2 tryptophan residues, with one out of five tryptophans more solvent-accessible in the ADAR2.RNA complex. PMID- 11479321 TI - Lack of complex I activity in human cells carrying a mutation in MtDNA-encoded ND4 subunit is corrected by the Saccharomyces cerevisiae NADH-quinone oxidoreductase (NDI1) gene. AB - The gene for the single subunit, rotenone-insensitive, and flavone-sensitive internal NADH-quinone oxidoreductase of Saccharomyces cerevisiae (NDI1) can completely restore the NADH dehydrogenase activity in mutant human cells that lack the essential mitochondrial DNA (mtDNA)-encoded subunit ND4. In particular, the NDI1 gene was introduced into the nuclear genome of the human 143B.TK(-) cell line derivative C4T, which carries a homoplasmic frameshift mutation in the ND4 gene. Two transformants with a low or high level of expression of the exogenous gene were chosen for a detailed analysis. In these cells the corresponding protein is localized in mitochondria, its NADH-binding site faces the matrix compartment as in yeast mitochondria, and in perfect correlation with its abundance restores partially or fully NADH-dependent respiration that is rotenone insensitive, flavone-sensitive, and antimycin A-sensitive. Thus the yeast enzyme has become coupled to the downstream portion of the human respiratory chain. Furthermore, the P:O ratio with malate/glutamate-dependent respiration in the transformants is approximately two-thirds of that of the wild-type 143B.TK(-) cells, as expected from the lack of proton pumping activity in the yeast enzyme. Finally, whereas the original mutant cell line C4T fails to grow in medium containing galactose instead of glucose, the high NDI1-expressing transformant has a fully restored capacity to grow in galactose medium. The present observations substantially expand the potential of the yeast NDI1 gene for the therapy of mitochondrial diseases involving complex I deficiency. PMID- 11479322 TI - Structural integrity of histone H2B in vivo requires the activity of protein L isoaspartate O-methyltransferase, a putative protein repair enzyme. AB - Protein L-isoaspartate O-methyltransferase (PIMT) is postulated to repair beta aspartyl linkages (isoaspartyl (isoAsp)) that accumulate at certain Asp-Xaa and Asn-Xaa sites in association with protein aging and deamidation. To identify major targets of PIMT action we cultured rat PC12 cells with adenosine dialdehyde (AdOx), a methyltransferase inhibitor that promotes accumulation of isoAsp in vivo. Subcellular fractionation of AdOx-treated cells revealed marked accumulation of isoAsp in a 14-kDa nuclear protein. Gel electrophoresis and chromatography of nuclei (3)H-methylated in vitro by PIMT revealed this protein to be histone H2B. The isoAsp content of H2B in AdOx-treated cells was approximately 18 times that in control cells, although no isoAsp was seen in other core histones, regardless of treatment. To confirm the relevance and specificity of this effect, we measured isoAsp levels in histones from brains of PIMT knockout mice. IsoAsp was found at near stoichiometric levels in H2B extracted from knockout brains and was at least 80 times greater than that in H2B from normal mice. Little or no isoAsp was detected in H2A, H3, or H4 from mice of either genotype. Accumulation of isoAsp in histone H2B may disrupt normal gene regulation and contribute to the reduced life span that characterizes PIMT knockouts. In addition to disrupting protein function, isoAsp has been shown to trigger immunity against self-proteins. The propensity of H2B to generate isoAsp in vivo may help explain why this histone in particular is found as a major antigen in autoimmune diseases such as lupus erythematosus. PMID- 11479323 TI - The gene ygdP, associated with the invasiveness of Escherichia coli K1, designates a Nudix hydrolase, Orf176, active on adenosine (5')-pentaphospho-(5') adenosine (Ap5A). AB - ygdP, a gene associated with the invasion of brain microvascular endothelial cells by Escherichia coli K1 (Badger, J. L., Wass, C. A., and Kim, K. S. (2000) Mol. Microbiol. 36, 174-182), the primary Gram-negative bacterium causing meningitis in newborns, has been cloned and expressed in E. coli. The protein, YgdP, was purified to near homogeneity and identified as a member of the Nudix hydrolase subfamily of dinucleoside oligophosphate pyrophosphatases. It catalyzes the hydrolysis of diadenosine tetra-, penta-, and hexa-phosphates with a preference for diadenosine penta-phosphate, from which it forms ATP and ADP. The enzyme has a requirement for a divalent metal cation that can be met with Mg2+, Zn2+, or Mn2+ and, like most of the Nudix hydrolases, has an alkaline pH optimum between 8.5 and 9. This is the second identification of a gene associated with the invasiveness of a human pathogen as a member of the Nudix hydrolase subfamily of dinucleoside oligophosphate pyrophosphatases, and an examination of homologous proteins in other invasive bacteria suggests that this may be a common feature of cellular invasion. PMID- 11479324 TI - Using gene knockouts to investigate plant metabolism. AB - Arabidopsis functional genomics resources now make the isolation of knockout mutants in any gene of choice both realistic and increasingly straightforward. Coupled with the completion of the genome sequence, this reverse genetics approach provides a platform facilitating dramatic progress in our understanding of fundamental aspects of plant metabolism. Recent experience shows that knockouts of genes encoding enzymes of primary metabolism can produce mutants with clear and sometimes unexpected phenotypes. They can provide new information about old pathways. Specific functions for individual members of multigene families can be revealed. Knockouts of enzymes of undefined function can lead to the discovery of those functions, and the analysis of enzymes which have previously never been studied at the biochemical level offers the potential to reveal new pathways of plant metabolism. Furthermore, the mutants isolated provide the starting point for genetic modification experiments to determine exactly how metabolism fuels growth and development, so providing a rational basis for the future modification of plant productivity. PMID- 11479325 TI - In vitro Arabidopsis pollen germination and characterization of the inward potassium currents in Arabidopsis pollen grain protoplasts. AB - The focus of this study is to investigate the regulatory role of K(+) influx in Arabidopsis pollen germination and pollen tube growth. Using agar-containing media, in vitro methods for Arabidopsis pollen germination have been successfully established for the first time. The pollen germination percentage was nearly 75% and the average pollen tube length reached 135 microm after a 6 h incubation. A decrease in external K(+) concentration from 1 mM to 35 microM resulted in 30% inhibition of pollen germination and 40% inhibition of pollen tube growth. An increase in external K(+) concentration from 1 mM to 30 mM stimulated pollen tube growth but inhibited pollen germination. To study how K(+) influx is associated with pollen germination and tube growth, regulation of the inward K(+) channels in the pollen plasma membrane was investigated by conducting patch-clamp whole cell recording with pollen protoplasts. K(+) currents were first identified in Arabidopsis pollen protoplasts. The inward K(+) currents were insensitive to changes in cytoplasmic Ca(2+) but were inhibited by a high concentration of external Ca(2+). A decrease of external Ca(2+) concentration from 10 mM (control) to 1 mM had no significant effect on the inward K(+) currents, while an increase of external Ca(2+) concentration from 10 mM to 50 mM inhibited the inward K(+) currents by 46%. Changes in external pH significantly affected the magnitude, conductance, voltage-independent maximal conductance, and activation kinetics of the inward K(+) currents. The physiological importance of potassium influx mediated by the inward K(+)-channels during Arabidopsis pollen germination and tube growth is discussed. PMID- 11479326 TI - Ethylene induces cell death at particular phases of the cell cycle in the tobacco TBY-2 cell line. AB - It was examined whether ethylene induces programmed cell death in a cell cycle specific manner. Following synchronization of the tobacco TBY-2 cell line with aphidicolin and its subsequent removal, ethylene was injected into the head space of 300 cm(3) culture flasks at 0 h or 3.5 h later and cells were sampled for 26 h. There were significant increases in cell mortality at G(2)/M in both the 0 h and 3.5 h ethylene treatments, and for the latter treatment, another peak in S phase. The effect at G(2)/M was greater in the 3.5 h treatment, but was ameliorated by the simultaneous addition of silver nitrate (1.2 microM). In addition, the 3.5 h ethylene treatment resulted in a 1 h delay in the characteristic rise in the mitotic index following aphidicolin-induced synchrony. The addition of silver nitrate alone (1.2 microM), also delayed the entry of cells into mitosis but had no effect on cell cycle length compared with the controls (14 h throughout all treatments) but it induced a peak of mortality 2.5 h after its addition. Nuclear shrinkage was also a characteristic feature of dying cells at G(2)/M. Using Apoptag, an in situ apoptosis detection kit, nuclear DNA fragmentation was observed in the TBY-2 cells which were often isolated on the end of a filament of normal cells. In the 3.5 h ethylene treatment, a marked increase was noted in the percentage of such cells at the G(2)/M transition compared with the controls. Hence, the data show cell death occurring at a major phase transition of the cell cycle and the observations of nuclear shrinkage, isolation of dying cells and nuclear DNA fragmentation suggest a programmed mechanism of cell death exacerbated by ethylene treatment. PMID- 11479327 TI - Analysis of cell division parameters and cell cycle gene expression during the cultivation of Arabidopsis thaliana cell suspensions. AB - Arabidopsis thaliana cell suspension cultures were characterized for the first time in detail in terms of biomass accumulation, cell division rate and cell cycle phase durations. Subsequently, this model system was used to follow the transcription profile of key cell cycle genes during a complete cultivation cycle. According to the calculated changes in the relative division rate over time, the cell cycle genes could be classified into four groups based on their transcriptional expression pattern. These differential patterns of gene expression are discussed with respect to the putative roles of the different cell cycle genes in the division cycle. Analysis of protein levels showed that mRNA levels did not correlate with protein levels in all cases. Results obtained in other systems, such as BY-2 cell suspensions or plants, confirm that cell suspension cultures of A. thaliana are suitable for the analysis of cell cycle regulation. PMID- 11479328 TI - beta-Galactosidases with a lectin-like domain are expressed in strawberry. AB - Strawberry fruits (Fragaria x ananassa Duch.) undergo a marked softening during their ripening, and the process is accompanied by a release of free sugars with galactose among them. In this work total beta-galactosidase activity was measured in cell wall proteins from strawberry fruits at different developmental stages. Three full-length cDNAs (Fa beta gal1, Fa beta gal2 and Fa beta gal3, respectively) encoding different beta-galactosidases (EC 3.2.1.23) were isolated from a library representing red fruit transcripts. All of them could be detected both in fruits and in vegetative tissues. However, only Fa beta gal1 showed an increasing expression during the ripening stages up to a maximum in the red fruits, while the other two (Fa beta gal2 and Fa beta gal3) were mostly found in green fruits and became barely detectable during ripening proper. The three beta galactosidase-encoding cDNAs were expressed in the yeast Pichia pastoris, and it was thus possible to demonstrate that each of them encode a beta-galactosidase. The expression of the three beta-galactosidase genes appears to be down-regulated by auxin, as already observed for other ripening-related genes of the non climacteric strawberry. An unusual characteristic of two strawberry beta galactosidases (Fa beta gal1 and Fa beta gal2) is that at the C-terminus of the enzymes a domain is found which is structurally related to known animal peptides with a sugar-binding ability. PMID- 11479329 TI - A comparative study of glutathione and ascorbate metabolism during germination of Pinus pinea L. seeds. AB - The ascorbate and glutathione systems have been studied during the first stages of germination in orthodox seeds of the gymnosperm Pinus pinea L. (pine). The results indicate that remarkable changes in the content and redox balance of these metabolites occur in both the embryo and endosperm; even if with different patterns for the two redox pairs. Dry seeds are devoid of the ascorbate reduced form (ASC) and contain only dehydroascorbic acid (DHA). By contrast, glutathione is present both in the reduced (GSH) and in the oxidized (GSSG) forms. During imbibition the increase in ASC seems to be mainly caused by the reactivation of its biosynthesis. On the other hand, the GSH rise occurring during the first 24 h seems to be largely due to GSSG reduction, even if GSH biosynthesis is still active in the seeds. The enzymes of the ascorbate--glutathione cycle also change during germination, but in different ways. ASC peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2) activities progressively rise both in the embryo and in endosperm. These changes are probably required for counteracting production of reactive oxygen species caused by recovery of oxidative metabolism. The two enzymes involved in the ascorbate recycling, ascorbate free radical (AFR) reductase (EC 1.6.5.4) and DHA reductase (EC 1.8.5.1), show different behaviour: the DHA reductase activity decreases, while that of AFR reductase remains unchanged. The relationship between ascorbate and glutathione metabolism and their relevance in the germination of orthodox seeds are also discussed. PMID- 11479330 TI - Nitrogen storage and remobilization in Brassica napus L. during the growth cycle: nitrogen fluxes within the plant and changes in soluble protein patterns. AB - Oilseed rape (Brassica napus L.) is commonly grown for oil or bio-fuel production, while the seed residues can be used for animal feed. It can also be grown as a catch crop because of its efficiency in extracting mineral N from the soil profile. However, the N harvest index is usually low, due in part to a low ability to remobilize N from leaves and to the fall of N-rich leaves which allows a significant amount of N to return to the environment. In order to understand how N filling of pods occurs, experiments were undertaken to quantify N flows within the plant by (15)N labelling and to follow the changes in soluble protein profiles of tissues presumed to store and subsequently to remobilize N. Whereas N uptake increased as a function of growth, N uptake capacity decreased at flowering to a non-significant level during pod filling. However, large amounts of endogenous N were transferred from the leaves to the stems and to taproots which acted as a buffering storage compartment later used to supply the reproductive tissue. About 15% of the total N cycling through the plant were lost through leaf fall and 48%, nearly all of which had been remobilized from vegetative tissues, were finally recovered in the mature pods. SDS-PAGE analysis revealed that large amounts of a 23 kDa polypeptide accumulated in the taproots during flowering and was later fully hydrolysed. Its putative function of storage protein is further supported by the fact that when plants were grown at lower temperature, both flowering, its accumulation and further mobilization were delayed. The overall results are discussed in relation to plant strategies which optimize N cycling to reproductive sinks by means of buffering vegetative tissues such as stems and taproots. PMID- 11479331 TI - Translocation of amino acids in the xylem of apple (Malus domestica Borkh.) trees in spring as a consequence of both N remobilization and root uptake. AB - Nitrogen is remobilized from storage for the growth of Malus domestica leaves each spring. Seasonal patterns of N translocation in the xylem sap as a consequence of remobilization were determined in 2-year-old 'Golden delicious' trees grafted on M9 rootstocks. The trees were grown in sand culture and (15)NH(4)(15)NO(3) at 10.4 atom% abundance supplied during August-September. The following year no further N was supplied and destructive harvests were taken during bud burst and leaf growth to determine the patterns of N remobilization together with the isolation of xylem sap for an analysis of their amino acid profiles and (15)N enrichments by GC-MS. The concentration of amino acids in the xylem sap rose following bud burst, peaked at full bloom and then fell again during petal fall and fruit set. The peak in amino acid concentration corresponded with the period when the rate of N remobilization was the fastest. The majority of labelled N was recovered in Asn, Gln + Glu and Asp demonstrating that they were being translocated as a consequence of remobilization. In a second experiment, 8-year-old trees growing in an orchard were fertilized with N either in the autumn or spring. Xylem sap samples were collected in the spring and early summer and, by comparison with the amino acid profiles recovered in trees from both treatments, Asn was identified as the main compound translocated as a consequence of both remobilization and root uptake of N, although there was evidence that root uptake of N occurred later. The data are discussed in relation to quantifying the internal cycling of N in trees. PMID- 11479332 TI - Gibberellic acid and dwarfism effects on the growth dynamics of B73 maize (Zea mays L.) leaf blades: a transient increase in apoplastic peroxidase activity precedes cessation of cell elongation. AB - The relationship between apoplastic peroxidase (EC 1.11.1.7) activity and cessation of growth in maize (Zea mays L.) leaf blades was investigated by altering elongation zone length. Apoplastic peroxidase activity in the elongation and secondary cell wall deposition zones of elongating leaf blades of the maize inbred line B73 was used as a control and compared to leaves of the dwarf mutant D8-81127, a near-isogenic line of B73 unresponsive to gibberellins, and to leaves of B73 plants to which gibberellic acid (GA(3)) had been applied via root uptake. Elongation zone length was increased by treatment with GA(3) through an increase in cell number as well as increased final cell length. The shorter elongation zone of dwarf leaves occurred primarily through reduced final cell length. Although elongation zone length differed among dwarf, control, and GA(3)-treated leaf blades, in all three treatments a transient increase in apoplastic peroxidase activity preceded a reduction in the segmental elongation rate in leaves. A peroxidase isoenzyme with pI 7.0 occurred in the leaf elongation zone during growth deceleration in all three treatments, and its activity decreased as growth displaced tissue into the region of secondary cell wall deposition. Growth cessation for all treatments coincided with the first appearance of peroxidase isozymes with pIs of 5.6 and 5.7. Based on the activity of particular isozymes relative to growth and differentiation, the pI 7.0 isoenzyme is most likely to be involved in cessation of cell elongation, while isozymes with pIs 5.6 and 5.7 are likely to be active in lignification. PMID- 11479333 TI - Distribution and mobility of aluminium in an Al-accumulating plant, Fagopyrum esculentum Moench. AB - Buckwheat (Fagopyrum esculentum Moench. cv. Jianxi) accumulates high concentrations of Al in the leaves without showing any toxicity. To understand the accumulation mechanism of Al in buckwheat, the distribution and mobility of Al in buckwheat were investigated. Relatively long-term treatment (28 d) with Al led to a decrease in Al concentration from old to young leaves, while a short term (1 d) exposure to Al resulted in a uniform distribution of Al in the leaves. When the fourth leaf was wrapped inside a transparent plastic bag to suppress transpiration, the Al concentration of this leaf was only one-quarter of that in the corresponding leaf without wrapping. Within a leaf, the Al concentration at the margins was much higher than that in the centre. These results indicate that Al distribution in the leaves is controlled by both rate and duration of transpiration. The mobility of Al between old and new leaves was studied by first growing plants in a solution with Al, followed by culture in a solution without Al. The Al content in the two new leaves appeared after removal of external Al was very low, whereas that in the old leaves did not decrease but continued to increase. The increased Al content was found to be translocated from Al remaining in the roots. It is concluded that Al is not mobile once it is accumulated in the leaf. PMID- 11479334 TI - Primary sites of ozone-induced perturbations of photosynthesis in leaves: identification and characterization in Phaseolus vulgaris using high resolution chlorophyll fluorescence imaging. AB - High resolution imaging of chlorophyll a fluorescence was used to identify the sites at which ozone initially induces perturbations of photosynthesis in leaves of Phaseolus vulgaris. Leaves were exposed to 250 and 500 nmol mol(-1) ozone at a photosynthetically active photon flux density of 300 micromol m(-2) s(-1) for 3 h. Images of fluorescence parameters indicated that large decreases in both the maximum and operating quantum efficiencies of photosystem II had occurred in cells adjacent to stomata in the upper, but not lower, leaf surfaces. However, this treatment did not produce any significant changes in the maximum or operating quantum efficiencies of photosystem II in the leaves when estimated from fluorescence parameters measured with a conventional, integrating fluorometer. The localized decreases in photosystem II photochemical efficiencies were accompanied by an increase in the minimal fluorescence level, which is indicative of photoinactivation of photosystem II complexes and a decrease in stomatal conductance. Perturbations of photochemical efficiencies were not observed in cells associated with all of the stomata on the upper leaf surface or within cells distant from the upper leaf surface. It is concluded that ozone penetrates the leaf through stomata and initially damages only cells close to stomatal pores. PMID- 11479335 TI - Transformation of Lotus japonicus using the herbicide resistance bar gene as a selectable marker. AB - Transgenic plants of the model legume Lotus japonicus were regenerated by hypocotyl transformation using a bar gene as a selectable marker. The bar encodes for Phosphinothricin Acetyl Transferase that detoxifies phosphinothricin (PPT), the active ingredient of herbicides such as Ignite (AgrEvo) and Basta (Hoechst). Transgenic L. japonicus plants resistant to PPT were positive upon PCR by bar gene-specific primers. In 5 out of 7 independent lines tested, PPT resistance segregated as a single dominant allele indicating a single T-DNA insertion into the plant genome. All regenerated plants were fertile and void of visible somaclonal abnormalities contrary to 14% infertility when antibiotic selectable markers were used. The lack of somaclonal variation, ease of PPT application and low cost of PPT makes this protocol an attractive alternative for the regeneration of transgenic L. japonicus. The production of PPT herbicide resistant L. japonicus plants may have significant commercial applications in crop production. PMID- 11479336 TI - Genotypic differences in the presence of hairs on roots and gynophores of peanuts (Arachis hypogaea L.) and their significance for phosphorus uptake. AB - Root hairs substantially increase the surface area of plant roots with positive effects for phosphorus (P) uptake, but the ability of peanuts to form root hairs has been questioned. The aim was to examine hair development on roots and gynophores of a variety of peanut genotypes and to relate genotypic differences in hair formation to differences in P uptake. Five out of eighteen genotypes completely lacked hairs on both organs whereas others consistently developed hairs on roots and gynophores, although with considerable variation in hair density. The ability to form root hairs as well as root hair density concurred with the presence and density of hairs on gynophores, suggesting a possible connection between both developmental processes. The contribution of root hairs to P uptake was studied in three genotypes differing in hair density. The final amount of P taken up by roots did not differ between genotypes but two distinct P uptake strategies could be identified. The genotype lacking root hairs maintained P uptake due to the development of a large root system whereas densely covered roots of genotype 'Wasedairyu' were three times as efficient in extracting P from a P-deficient soil. Furthermore P uptake through gynophores contributed about 20% to the total P uptake of Wasedairyu but only insignificant amounts to other genotypes. The ability to form hairs on roots and gynophores can therefore be seen as an adaptation to low P availability and if combined with a large root system, could substantially increase the tolerance of peanuts to P deficiency. PMID- 11479337 TI - Heat-shock responses in two leguminous plants: a comparative study. AB - Relative growth rates, basal and acclimated thermotolerance, membrane damage, fluorescence emission, and relative levels of free and conjugated ubiquitin and HSP70 were compared after 2 h of treatment at different temperatures between Prosopis chilensis and Glycine max (soybean), cv. McCall, to evaluate if the thermotolerance of these two plants was related to levels of accumulation of heat shock proteins. Seedlings of P. chilensis germinated at 25 degrees C and at 35 degrees C and grown at temperatures above germination temperature showed higher relative growth than soybean seedlings treated under the same conditions. The lethal temperature of both species was 50 degrees C after germination at 25 degrees C. However, they were able to grow at 50 degrees C after germination at 35 degrees C. Membrane damage determinations in leaves showed that P. chilensis has an LT(50) 6 degrees C higher than that of soybean. There were no differences in the quantum yield of photosynthesis (F(v)/F(m)), between both plants when the temperatures were raised. P. chilensis showed higher relative levels of free ubiquitin, conjugated ubiquitin and HSP70 than soybean seedlings when the temperatures were raised. Time-course studies of accumulation of these proteins performed at 40 degrees C showed that the relative accumulation rates of ubiquitin, conjugated ubiquitin and HSP70 were higher in P. chilensis than in soybean. In both plants, free ubiquitin decreased during the first 5 min and increased after 30 min of heat shock, conjugated ubiquitin increased after 30 min and HSP70 began to increase dramatically after 20 min of heat shock. From these data it is concluded that P. chilensis is more tolerant to acute heat stress than soybean. PMID- 11479338 TI - Hydrogen peroxide induces programmed cell death features in cultured tobacco BY-2 cells, in a dose-dependent manner. AB - Active oxygen species (AOS), especially hydrogen peroxide, play a critical role in the defence of plants against invading pathogens and in the hypersensitive response (HR). This is characterized by the induction of a massive production of AOS and the rapid appearance of necrotic lesions is considered as a programmed cell death (PCD) process during which a limited number of cells die at the site of infection. This work was aimed at investigating the mode of cell death observed in cultures of BY-2 tobacco cells exposed to H(2)O(2). It was shown that H(2)O(2) is able to induce various morphological cell death features in cultured tobacco BY-2 cells. The hallmarks of cell death observed with fluorescent and electron microscopy differed greatly with the amount of H(2)O(2) added to the cell culture. The appearance of nuclear fragmentation similar to 'apoptotic bodies' associated with a fragmentation of the nuclear DNA into small fragments appear for almost 18% of the cells treated with 12.5 mM H(2)O(2). The early stages of the induction of this PCD process consisted in cell shrinkage and chromatin condensation at the periphery of the nucleus. Above 50 mM, H(2)O(2) induces high necrotic cell death. These data suggest that H(2)O(2)-induced cell damage is associated with the induction of various cell death processes that could be involved differently in plant defence reactions. PMID- 11479339 TI - Boron supply into wheat (Triticum aestivum L. cv. Wilgoyne) ears whilst still enclosed within leaf sheaths. AB - The present study investigates whether there is significant remobilization of (10)B previously loaded in the flag and penultimate leaves into the young, actively growing ear enclosed within the sheaths of flag and penultimate leaves. It also explores whether B transport into the enclosed ear declines when air humidity in the shoot canopy increases. After 5 d (10)B labelling during the period from early to full emergence of the flag leaf, the plants were transferred into nutrient solutions containing either 10 microM (11)B or no added B for 3 d. Regardless of the subsequent B supply levels to the roots, (10)B contents in the ear continued to increase by up to 5-fold 3 d after the end of (10)B supply in the nutrient solution. During these 3 d, the ear experienced a rapid increase in biomass. However, the majority of B in the ear during the 3 d treatment period was from the newly acquired (11)B from root uptake, rather than retranslocation of (10)B previously deposited in the leaves. By comparing the relative distribution of (10)B, Rb (xylem-to-phloem transfer marker) and Sr (xylem-marker) in the ear and the flag leaf, the distribution of (10)B resembled that of Rb more than Sr. Canopy cover treatment greatly suppressed leaf transpiration and decreased the amount of newly acquired (10)B in the flag leaf and the ear, but not in the upper stem segments. The results suggest that whilst the young ear was still fully enclosed within the leaf sheaths without any significant transpiration activity, B transport into the ear is predominantly dependent on the long-distance B transport in the xylem driven by leaf transpiration and, therefore, on concurrent B uptake from the roots. PMID- 11479340 TI - ZMPP2, a novel type-2C protein phosphatase from maize. AB - A cDNA clone was selected as a candidate for the catalytic subunit of phospho pyruvate dehydrogenase phosphatase (PDP) by screening a Zea mays expressed sequence tag database with the bovine PDP deduced amino acid sequence. Both strands of the cDNA were completely sequenced. The maize clone contains an open reading frame of 1098 base pairs that encodes a polypeptide of 40 127 Da, ZMPP2. The deduced amino acid sequence of ZMPP2 contains the five PP2C signature domains, as does PDP. However, the expression pattern of ZMPP2, determined by reverse transcriptase-polymerase chain reaction, was different from those of the maize pyruvate dehydrogenase E1 alpha subunit and pyruvate dehydrogenase kinase. Additionally, the predicted subcellular location of ZMPP2 is cytoplasmic, while the pyruvate dehydrogenase complex, regulated by reversible phosphorylation, is mitochondrial. Thus, ZMPP2 is a PP2C-type protein phosphatase related to but distinct from PDP. PMID- 11479341 TI - The relationship between cAMP, Ca(2)+, and transport of CFTR to the plasma membrane. AB - The mechanism whereby cAMP stimulates Cl(-) flux through CFTR ion channels in secretory epithelia remains controversial. It is generally accepted that phosphorylation by cAMP-dependent protein kinase increases the open probability of the CFTR channel. A more controversial hypothesis is that cAMP triggers the translocation of CFTR from an intracellular pool to the cell surface. We have monitored membrane turnover in Calu-3 cells, a cell line derived from human airway submucosal glands that expresses high levels of CFTR using membrane capacitance and FM1-43 fluorescence measurements. Using a conventional capacitance measurement technique, we observe an apparent increase in membrane capacitance in most cells that exhibit an increase in Cl(-) current. However, after we carefully correct our recordings for changes in membrane conductance, the apparent changes in capacitance are eliminated. Measurements using the fluorescent membrane marker FM1-43 also indicate that no changes in membrane turnover accompany the activation of CFTR. Robust membrane insertion can be triggered with photorelease of caged Ca(2)+ in Calu-3 cells. However, no increase in Cl(-) current accompanies Ca(2)+-evoked membrane fusion. We conclude that neither increases in cAMP or Ca(2)+ lead to transport of CFTR to the plasma membrane in Calu-3 cells. In addition, we conclude that membrane capacitance measurements must be interpreted with caution when large changes in membrane conductance occur. PMID- 11479342 TI - A kinetic analysis of calcium-triggered exocytosis. AB - Although the relationship between exocytosis and calcium is fundamental both to synaptic and nonneuronal secretory function, analysis is problematic because of the temporal and spatial properties of calcium, and the fact that vesicle transport, priming, retrieval, and recycling are coupled. By analyzing the kinetics of sea urchin egg secretory vesicle exocytosis in vitro, the final steps of exocytosis are resolved. These steps are modeled as a three-state system: activated, committed, and fused, where interstate transitions are given by the probabilities that an active fusion complex commits (alpha) and that a committed fusion complex results in fusion, p. The number of committed complexes per vesicle docking site is Poisson distributed with mean n. Experimentally, p and n increase with increasing calcium, whereas alpha and the pn ratio remain constant, reducing the kinetic description to only one calcium-dependent, controlling variable, n. On average, the calcium dependence of the maximum rate (R(max)) and the time to reach R(max) (T(peak)) are described by the calcium dependence of n. Thus, the nonlinear relationship between the free calcium concentration and the rate of exocytosis can be explained solely by the calcium dependence of the distribution of fusion complexes at vesicle docking sites. PMID- 11479343 TI - Dynamic interaction of S5 and S6 during voltage-controlled gating in a potassium channel. AB - A gain-of-function mutation in the Caenorhabditis elegans exp-2 K(+)-channel gene is caused by a cysteine-to-tyrosine change (C480Y) in the sixth transmembrane segment of the channel (Davis, M.W., R. Fleischhauer, J.A. Dent, R.H. Joho, and L. Avery. 1999. Science. 286:2501-2504). In contrast to wild-type EXP-2 channels, homotetrameric C480Y mutant channels are open even at -160 mV, explaining the lethality of the homozygous mutant. We modeled the structure of EXP-2 on the 3-D scaffold of the K(+) channel KcsA. In the C480Y mutant, tyrosine 480 protrudes from S6 to near S5, suggesting that the bulky side chain may provide steric hindrance to the rotation of S6 that has been proposed to accompany the open closed state transitions (Perozo, E., D.M. Cortes, and L.G. Cuello. 1999. Science. 285:73-78). We tested the hypothesis that only small side chains at position 480 allow the channel to close, but that bulky side chains trap the channel in the open state. Mutants with small side chain substitutions (Gly and Ser) behave like wild type; in contrast, bulky side chain substitutions (Trp, Phe, Leu, Ile, Val, and His) generate channels that conduct K(+) ions at potentials as negative as -120 mV. The side chain at position 480 in S6 in the pore model is close to and may interact with a conserved glycine (G421) in S5. Replacement of G421 with bulky side chains also leads to channels that are trapped in an active state, suggesting that S5 and S6 interact with each other during voltage-dependent open-closed state transitions, and that bulky side chains prevent the dynamic changes necessary for permanent channel closing. Single-channel recordings show that mutant channels open frequently at negative membrane potentials indicating that they fail to reach long-lasting, i.e., stable, closed states. Our data support a "two-gate model" with a pore gate responsible for the brief, voltage-independent openings and a separately located, voltage-activated gate (Liu, Y., and R.H. Joho. 1998. Pflugers Arch. 435:654 661). PMID- 11479344 TI - Molecular architecture of the voltage-dependent Na channel: functional evidence for alpha helices in the pore. AB - The permeation pathway of the Na channel is formed by asymmetric loops (P segments) contributed by each of the four domains of the protein. In contrast to the analogous region of K channels, previously we (Yamagishi, T., M. Janecki, E. Marban, and G. Tomaselli. 1997. Biophys. J. 73:195-204) have shown that the P segments do not span the selectivity region, that is, they are accessible only from the extracellular surface. The portion of the P-segment NH(2)-terminal to the selectivity region is referred to as SS1. To explore further the topology and functional role of the SS1 region, 40 amino acids NH(2)-terminal to the selectivity ring (10 in each of the P segments) of the rat skeletal muscle Na channel were substituted by cysteine and expressed in tsA-201 cells. Selected mutants in each domain could be blocked with high affinity by externally applied Cd(2)+ and were resistant to tetrodotoxin as compared with the wild-type channel. None of the externally applied sulfhydryl-specific methanethiosulfonate reagents modified the current through any of the mutant channels. Both R395C and R750C altered ionic selectivity, producing significant increases in K(+) and NH(4)(+) currents. The pattern of side chain accessibility is consistent with a pore helix like that observed in the crystal structure of the bacterial K channel, KcsA. Structure prediction of the Na channel using the program PHDhtm suggests an alpha helix in the SS1 region of each domain channel. We conclude that each of the P segments undergoes a hairpin turn in the permeation pathway, such that amino acids on both sides of the putative selectivity filter line the outer mouth of the pore. Evolutionary conservation of the pore helix motif from bacterial K channels to mammalian Na channels identifies this structure as a critical feature in the architecture of ion selective pores. PMID- 11479345 TI - Molecular regions controlling the activity of CNG channels. AB - The alpha subunits of CNG channels of retinal photoreceptors (rod) and olfactory neurons (olf) are proteins that consist of a cytoplasmic NH(2) terminus, a transmembrane core region (including the segments S1-S6), and a cytoplasmic COOH terminus. The COOH terminus contains a cyclic nucleotide monophosphate binding domain NBD) that is linked by the C-linker (CL) to the core region. The binding of cyclic nucleotides to the NBD promotes channel opening by an allosteric mechanism. We examined why the sensitivity to cGMP is 22 times higher in olf than in rod by constructing chimeric channels and determining the [cGMP] causing half maximum channel activity (EC(50)). The characteristic difference in the EC(50) value between rod and olf was introduced by the NH(2) terminus and the core-CL region, whereas the NBD showed a paradoxical effect. The difference of the free energy difference Delta(DeltaG) was determined for each of these three regions with all possible combinations of the other two regions. For rod regions with respect to corresponding olf regions, the open channel conformation was destabilized by the NH(2) terminus (Delta(DeltaG) = -1.0 to -2.0 RT) and the core CL region (Delta(DeltaG) = -2.0 to -2.9 RT), whereas it was stabilized by the NBD (Delta(DeltaG) = 0.3 to 1.1 RT). The NH(2) terminus deletion mutants of rod and olf differed by Delta(DeltaG) of only 0.9 RT, whereas the wild-type channels differed by the much larger value of 3.1 RT. The results show that in rod and olf, the NH(2) terminus, the core-CL region, and the NBD differ by characteristic Delta(DeltaG) values that do not depend on the specific composition of the other two regions and that the NH(2) terminus generates the main portion of Delta(DeltaG) between the wild-type channels. PMID- 11479346 TI - Site-directed spin-labeling analysis of reconstituted Mscl in the closed state. AB - The mechanosensitive channel from Escherichia coli (Eco-MscL) responds to membrane lateral tension by opening a large, water-filled pore that serves as an osmotic safety valve. In an attempt to understand the structural dynamics of MscL in the closed state and under physiological conditions, we have performed a systematic site-directed spin labeling study of this channel reconstituted in a membrane bilayer. Structural information was derived from an analysis of probe mobility, residue accessibility to O(2) or NiEdda and overall intersubunit proximity. For the majority of the residues studied, mobility and accessibility data showed a remarkable agreement with the Mycobacterium tuberculosis crystal structure, clearly identifying residues facing the large water-filled vestibule at the extracellular face of the molecule, the narrowest point along the permeation pathway (residues 21-26 of Eco-MscL), and the lipid-exposed residues in the peripheral transmembrane segments (TM2). Overall, the present dataset demonstrates that the transmembrane regions of the MscL crystal structure (obtained in detergent and at low pH) are, in general, an accurate representation of its structure in a membrane bilayer under physiological conditions. However, significant differences between the EPR data and the crystal structure were found toward the COOH-terminal end of TM2. PMID- 11479347 TI - Extracellular blockade of K(+) channels by TEA: results from molecular dynamics simulations of the KcsA channel. AB - TEA is a classical blocker of K(+) channels. From mutagenesis studies, it has been shown that external blockade by TEA is strongly dependent upon the presence of aromatic residue at Shaker position 449 which is located near the extracellular entrance to the pore (Heginbotham, L., and R. MacKinnon. 1992. Neuron. 8:483-491). The data suggest that TEA interacts simultaneously with the aromatic residues of the four monomers. The determination of the 3-D structure of the KcsA channel using X-ray crystallography (Doyle, D.A., J.M. Cabral, R.A. Pfuetzner, A. Kuo, J.M. Gulbis, S.L. Cohen, B.T. Chait, and R. MacKinnon. 1998. Science. 280:69-77) has raised some issues that remain currently unresolved concerning the interpretation of these observations. In particular, the center of the Tyr82 side chains in KcsA (corresponding to position 449 in Shaker) forms a square of 11.8-A side, a distance which is too large to allow simultaneous interactions of a TEA molecule with the four aromatic side chains. In this paper, the external blockade by TEA is explored by molecular dynamics simulations of an atomic model of KcsA in an explicit phospholipid bilayer with aqueous salt solution. It is observed, in qualitative accord with the experimental results, that TEA is stable when bound to the external side of the wild-type KcsA channel (with Tyr82), but is unstable when bound to a mutant channel in which the tyrosine residue has been substituted by a threonine. The free energy profile of TEA relative to the pore is calculated using umbrella sampling simulations to characterize quantitatively the extracellular blockade. It is found, in remarkable agreement with the experiment, that the TEA is more stably bound by 2.3 kcal/mol to the channel with four tyrosine residues. In the case of the wild type KcsA channel, TEA (which has the shape of a flattened oblate spheroid) acts as an ideal plug blocking the pore. In contrast, it is considerably more off centered and tilted in the case of the mutant channel. The enhanced stability conferred by the tyrosine residues does not arise from Pi-cation interactions, but appears to be due to differences in the hydration structure of the TEA. Finally, it is shown that the experimentally observed voltage dependence of TEA block, which is traditionally interpreted in terms of the physical position of the TEA along the axis of the pore, must arise indirectly via coupling with the ions in the pore. PMID- 11479348 TI - Is cystic fibrosis lung disease caused by abnormal ion composition or abnormal volume? PMID- 11479349 TI - The relative roles of passive surface forces and active ion transport in the modulation of airway surface liquid volume and composition. AB - Two hypotheses have been proposed recently that offer different views on the role of airway surface liquid (ASL) in lung defense. The "compositional" hypothesis predicts that ASL [NaCl] is kept low (<50 mM) by passive forces to permit antimicrobial factors to act as a chemical defense. The "volume" hypothesis predicts that ASL volume (height) is regulated isotonically by active ion transport to maintain efficient mechanical mucus clearance as the primary form of lung defense. To compare these hypotheses, we searched for roles for: (1) passive forces (surface tension, ciliary tip capillarity, Donnan, and nonionic osmolytes) in the regulation of ASL composition; and (2) active ion transport in ASL volume regulation. In primary human tracheobronchial cultures, we found no evidence that a low [NaCl] ASL could be produced by passive forces, or that nonionic osmolytes contributed substantially to ASL osmolality. Instead, we found that active ion transport regulated ASL volume (height), and that feedback existed between the ASL and airway epithelia to govern the rate of ion transport and volume absorption. The mucus layer acted as a "reservoir" to buffer periciliary liquid layer height (7 microm) at a level optimal for mucus transport by donating or accepting liquid to or from the periciliary liquid layer, respectively. These data favor the active ion transport/volume model hypothesis to describe ASL physiology. PMID- 11479350 TI - Economics and ethics in health care. PMID- 11479351 TI - Ethics and health care 'underfunding'. PMID- 11479352 TI - A 'fair innings' for efficiency in health services? AB - This paper reviews the severe visual focus problems of health economists-they have developed a one-sided fixation with equity issues, neglecting the efficiency agenda. The problems of meeting need are not just about access--they will vary with cost and supply. Economists in fact developed a more balanced agenda in the 1970s but have failed to follow it up. The paper defines the triple nationalisation of the National Health Service (NHS), and presents evidence that pluralism, using the purchaser/provider split, has become more efficient in long term care, home care and services for people with learning difficulties. Health economics has failed to explore options for using the dynamic forces of choice and competition for improving health services. The paper makes the case for a decision rule that "we should seek the process of health care service supply and development which maximises the delivery of high quality, lower cost user relevant services". PMID- 11479353 TI - Ethics, economics, and public financing of health care. AB - There is a wide variety of ethical arguments for public financing of health care that share a common structure built on a series of four logically related propositions regarding: (1) the ultimate purpose of a human life or human society; (2) the role of health and its distribution in society in advancing this ultimate purpose; (3) the role of access to or utilisation of health care in maintaining or improving the desired level and distribution of health among members of society, and (4) the role of public financing in ensuring the ethically justified access to and utilisation of health care by members of society. This paper argues that economics has much to contribute to the development of the ethical foundations for publicly financed health care. It focuses in particular on recent economic work to clarify the concepts of access and need and their role in analyses of the just distribution of health care resources, and on the importance of economic analysis of health care and health care insurance markets in demonstrating why public financing is necessary to achieve broad access to and utilisation of health care services. PMID- 11479354 TI - Individual autonomy and state involvement in health care. AB - This article examines the ethical basis for government involvement in health care. It first provides the case for individual autonomy, focusing on the justifications--particularly ethical ones--for allowing individuals to make their own choices in health care, and to control more of their own resources in doing so. Next, it provides the opposite case--for abridging individual autonomy, and in particular, for redistributing resources from those who are well off to those who are not. The overriding reason for favouring the latter case, which trumps the notion of individual autonomy, is to ensure that individuals who are at a disadvantage have an equal probability of attaining good health. PMID- 11479355 TI - Analysing health outcomes. AB - If we cross-classify the absolutist-consequentialist distinction with an intuitive-analytical one we can see that economists probably attract the hostility of those in the other three cells as a result of being analytical consequentialists, as much as because of their concern with "costs". Suggesting that some sources of utility (either "outcome" or "process" in origin) are to be regarded as rights cannot, says the analytical consequentialist, overcome the fact that fulfilling and respecting rights is a resource-consuming activity, one that will inevitably have consequences, in resource-constrained situations, for the fulfillment of the rights of others. Within the analytical consequentialist framework QALY-type measures of health outcome have the unique advantage of allowing technical and allocative efficiency to be addressed simultaneously, while differential weighting of QALYs accruing to different groups means that efficiency and equity can be merged into the necessary single maximand. But what if such key concepts of the analytical consequentialist are not part of the discursive equipment of others? Are they to be disqualified from using them on this ground? Is it ethical for intuition to be privileged in ethical discourse, or is the analyst entitled to "equal opportunities" in the face of "analysisism", the cognitive equivalent of "racism" and "sexism"? PMID- 11479356 TI - How economics could extend the scope of ethical discourse. AB - Ethical discourse is typically inconclusive, and with good reason. But this inconclusiveness is a distinct disadvantage when it comes to helping publicly accountable policy-makers in the health care system provide an ethical justification for their decisions. It is suggested that instead of ending with platitudinous statements such as that a balance has to be struck between the rival ethical considerations, empirical research should be undertaken to elicit the quantitative trade-offs that the affected general public would be prepared to accept when striking this balance. In the expected absence of any consensus, it is further suggested that the views of the median person be taken as the best approximation to the group view. Finally it is argued that, far from this quantitative approach lacking humanity by treating individuals as "mere statistics", it shows greater compassion than the proponents of those approaches whose fellow feeling can only be stirred by information pertaining to identified individuals. PMID- 11479357 TI - Ethics and geographical equity in health care. AB - Important variations in access to health care and health outcomes are associated with geography, giving rise to profound ethical concerns. This paper discusses the consequences of such concerns for the allocation of health care finance to geographical regions. Specifically, it examines the ethical drivers underlying capitation systems, which have become the principal method of allocating health care finance to regions in most countries. Although most capitation systems are based on empirical models of health care expenditure, there is much debate about which needs factors to include in (or exclude from) such models. This concern with legitimate and illegitimate drivers of health care expenditure reflects the ethical concerns underlying the geographical distribution of health care finance. PMID- 11479358 TI - Economics, health and development: some ethical dilemmas facing the World Bank and the international community. AB - The World Bank is committed to "work[ing] with countries to improve the health, nutrition and population outcomes of the world's poor, and to protect[ing] the population from the impoverishing effects of illness, malnutrition and high fertility".(1) Ethical issues arise in the interpretation of these objectives and in helping countries formulate strategies and policies. It is these ethical issues--which are often not acknowledged by commentators--that are the subject of this paper. It asks why there should be a focus on the poor, and explores the link between improving the health of the poor, and reducing health inequalities between the poor and better-off. It discusses difficult ethical issues at both the global level (including debt relief and the link between country ownership and donor commitment) and the country level (including user fees and whether providing assistance to the non-poor may in the long run be a way of helping the poor). PMID- 11479359 TI - Should physicians be gatekeepers of medical resources? AB - Physicians have an ethical responsibility to their patients to offer the best available medical care. This responsibility conflicts with their role as gatekeepers of the limited health care resources available for all patients collectively. It is ethically untenable to expect doctors to face this trade-off during each patient encounter; the physician cannot be expected to compromise the wellbeing of the patient in the office in favour of anonymous patients elsewhere. Hence, as in other domains of public policy where individual and collective interests conflict, some form of collective solution is required. Collective solutions may take the form of placing explicit resource constraints on resources available to physicians, or clinical practice guidelines that recognise cost effective care as acceptable. Such solutions will be politically and ethically sustainable only if patients as citizens of the larger population accept the need for rationing of limited resources in health care. PMID- 11479360 TI - Equity - some theory and its policy implications. AB - This essay seeks to characterise the essential features of an equitable health care system in terms of the classical Aristotelian concepts of horizontal and vertical equity, the common (but ill-defined) language of "need" and the economic notion of cost-effectiveness as a prelude to identifying some of the more important issues of value that policy-makers will have to decide for themselves; the characteristics of health (and what determines it) that can cause policy to be ineffective (or have undesired consequences); the information base that is required to support a policy directed at securing greater equity, and the kinds of research (theoretical and empirical) that are needed to underpin such a policy. PMID- 11479361 TI - Energizing epithelial transport with the vacuolar H(+)-ATPase. AB - The Ussing model has long provided the conceptual foundation for understanding epithelial transport mechanisms energized by the Na(+)-K(+)-ATPase. Plasma membranes may also use the vacuolar (V-type) H(+)-ATPase as the primary energy source of membrane and epithelial transport. A pure electrogenic pump, the V-type H(+)-ATPase energizes not only membranes it inhabits but also other transport pathways via electrical coupling. PMID- 11479363 TI - The intrarenal endothelin system and hypertension. AB - The kidney is both a source of endothelin (ET) generation and an important target organ of this peptide. The highest concentrations of ET-1 in the body exist in the renal medulla, where it mediates natriuretic and diuretic effects through the ET(B) receptor subtype. It is proposed that aberrations in the renal ET system may lead to sodium and water retention and subsequently to the development of hypertension. PMID- 11479364 TI - The glutamine/glutamate couplet and cellular function. AB - All cells require glutamine as a nitrogen donor as well as an energy source for cell-specific functions. Understanding how glutamine utilization is metered to these demands is fundamental to basic cell processes as well as to therapeutic manipulation of regulatory mechanisms. The regulatory role of the glutamine/glutamate couplet in cellular function is illustrated for acid-base homeostasis and for production of the extracellular matrix. PMID- 11479365 TI - ClC chloride channels in epithelia: recent progress and remaining puzzles. AB - ClC chloride channels are widely expressed in epithelia. Recent insights into the roles of specific ClC channels have emerged from molecular and immunolocalization studies, mouse knockout models, and the linkage of mutations of these channels to the human hereditary diseases Bartter's syndrome and Dent's disease. PMID- 11479366 TI - CFTR: interacting with everything? AB - More than 1,300 different mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) are the cause for cystic fibrosis. CFTR is in charge of proper secretion and absorption of electrolytes, and thus the disease is characterized by defective epithelial Cl(-) secretion and enhanced Na(+) absorption. Recent studies show that CFTR interacts with other proteins via PDZ domains. PMID- 11479367 TI - Molecular recognition by induced fit: how fit is the concept? AB - Induced fit explains why biomolecules can bind together even if they are not optimized for binding. However, induced fit can lead to a kinetic bottleneck and does not describe every interaction in the absence of prior complementarity. Preselection of a fitting conformer is an alternative to induced fit. PMID- 11479368 TI - Sex and salt hormones: rapid effects in epithelia. AB - Recent evidence points to protein kinase C isoforms as highly specific receptors for aldosterone and estradiol in epithelia. The end targets of the kinase activation are Na(+)/H(+) exchange and K(+) and Ca(2+) channels. The physiological role of the nongenomic response is to increase electrolyte absorption and inhibit secretion in pluripotential epithelia. PMID- 11479369 TI - Synaptic transmission with the glia. AB - For decades, scientists thought that all of the missing secrets of brain function resided in neurons. However, a wave of new findings indicates that glial cells, formerly considered mere supporters and subordinate to neurons, participate actively in synaptic integration and processing of information in the brain. PMID- 11479370 TI - Reactive species in viral pneumonitis: lessons from animal models. AB - Recent evidence suggests that pneumonitis induced by many important human viral pathogens may result from exuberant generation of reactive species by inflammatory cells in response to infection. This review summarizes current evidence from animal model studies regarding the beneficial (antiviral) and harmful (tissue-damaging) effects of reactive species for the host. PMID- 11479371 TI - Antibody-based targeting of angiogenesis. AB - The selective targeting of neovasculature opens new avenues for the diagnosis and therapy of angiogenesis-related diseases such as cancer, blinding ocular disorders, and rheumatoid arthritis. Here we review recent advances in the identification of markers of angiogenesis as well as in the isolation and use of antibodies (and their derivatives) for the in vivo targeting of both tumoral and nontumoral neovasculature. PMID- 11479372 TI - Oxygen sensing: "hydroxy" translates "oxy". PMID- 11479373 TI - Identification of plant cytokinin biosynthetic enzymes as dimethylallyl diphosphate:ATP/ADP isopentenyltransferases. AB - It has been believed that the key step in cytokinin biosynthesis is the addition of a 5-carbon chain to the N(6) of AMP. To identify cytokinin biosynthesis enzymes that catalyze the formation of the isopentenyl side chain of cytokinins, the Arabidopsis genomic sequence was searched for genes that could code for isopentenyltransferases. This resulted in the identification of nine putative genes for isopentenyltransferases. One of these, AtIPT4, was subjected to detailed analysis. Overexpression of AtIPT4 caused cytokinin-independent shoot formation on calli. As shoot formation on calli normally occurs only when cytokinins are applied, it suggested that this gene product catalyzed cytokinin biosynthesis in plants. Recombinant AtIPT4 catalyzed the transfer of an isopentenyl group from dimethylallyl diphosphate to the N(6) of ATP and ADP, but not to that of AMP. AtIPT4 did not exhibit the DMAPP:tRNA isopentenyltransferase activity. These results indicate that cytokinins are, at least in part, synthesized from ATP and ADP in plants. PMID- 11479374 TI - MIP genes are down-regulated under drought stress in Nicotiana glauca. AB - Water flux across cell membranes has been shown to occur not only through the lipid bilayer, but also through aquaporins, which are members of the major intrinsic protein (MIP) super-family of channel proteins. Aquaporins greatly increase the membrane permeability for water, but may also be regulated, allowing cellular control over the rate of water influx/efflux. Water flux is crucial for stomatal opening and closing, but little is known about the role that aquaporins play in stomatal physiology. Our initial goal was to isolate and characterize the MIP genes expressed in guard cells of the model plant, Nicotiana glauca. Degenerate oligonucleotides corresponding to amino acid sequences conserved in tonoplast intrinsic proteins (TIPs) or plasma membrane intrinsic proteins (PIPs) were used to amplify portions of MIP genes by RT-PCR. These PCR products were used as probes in screening a N. glauca guard cell cDNA library. We isolated three clones (NgMIP1, NgMIP2 and NgMIP3) homologous to TIPs and two clones (NgMIP4 and NgMIP5) homologous to PIPs. All of the MIP genes we characterized displayed highest levels of mRNA accumulation in roots or stems, with lower levels of expression in mesophyll cells and whole leaves, and lowest transcript accumulation in guard cell RNA. Interestingly, the accumulation of transcripts arising from NgMIP2, NgMIP3 and NgMIP4 diminished dramatically in drought stressed plants. This down-regulation of MIP gene expression may result in reduced membrane water permeability and may encourage cellular water conservation during periods of dehydration stress. PMID- 11479375 TI - Changes in growth and structure of pea primary roots (Pisum sativum L. cv. Alaska) as a result of sudden flooding. AB - Pea (Pisum sativum L. cv. Alaska) primary roots were exposed to flooding after growth for 4 or 5 d at 25 degrees C under relatively dry conditions. Flooding after 4 d growth reduced, but did not stop, primary root growth, and cavities caused by degradation of central vascular cells were typically found from 10-60 mm from the tips. Flooding after 5 d stopped primary root growth and caused cell death in the tips, and vascular cavities formed that typically were 20-60 mm from the tips of the roots. Degradation of root tip cells in 5-day-roots was very rapid and began in the elongation zone and later in the apical zone. Root tips discolored, narrowed or curled before growth arrest. The mitotic indices of 5-day root tips were suppressed by the flooding treatment. A few mitotic figures were observed in roots treated with flooding after 4 d growth. Affected cells had condensed nuclei, but cytoplasms appeared to be normal in the early stages of cell degradation. Later these cells became very vacuolated. The relationship of flooding to root growth, vascular cavity formation, and the morphology of pea primary roots is described with regard to the ability to resist flooding stress. PMID- 11479376 TI - Mutational analysis of regulatory cis-acting elements for the transcriptional activation of the dmsCBA operon in Rhodobacter sphaeroides f. sp. denitrificans. AB - Four direct repeats of a 10-nt sequence, called dms boxes, are located upstream of the dmsCBA operon encoding dimethyl sulfoxide (DMSO) reductase in Rhodobacter sphaeroides f. sp. denitrificans IL106. Two dms boxes 1 and 2 have been shown to be binding sites of DmsR protein, a response regulator of a two-component system involved in the anaerobic induction by DMSO of DMSO reductase synthesis. In this study, functions of four dms boxes in the transcriptional regulation of the dmsCBA operon were investigated. The transcription start site of the dmsCBA genes was identified at the distance of 23 nt downstream of the closest dms box 4. Expression of the dmsC-lacZ gene fusion which included the dmsCBA promoter region containing the dms boxes was examined and its anaerobic induction by DMSO and DmsR-dependency were demonstrated in the phototroph. The examination with nucleotide substitutions in the four respective dms boxes showed that the set of four dms boxes is required for the dmsCBA operon activation. Moreover, the importance of the nucleotide sequence of TTCAC in dms box 4 and of A at the center in dms box 1 was significantly shown. These facts suggest that the pentad nucleotides TTCAC and TTAAC in the dms boxes serve as cis-acting elements in the transcriptional activation of the dmsCBA operon. PMID- 11479377 TI - Isolation of the protease component of maize cysteine protease-cystatin complex: release of cystatin is not crucial for the activation of the cysteine protease. AB - The maize cysteine protease complex, which required SDS for its activation in vitro, is a 179 kDa trimeric complex (P-I)3 of a cysteine protease (P) [EC 3.4.22] and a cysteine protease inhibitor (I), cystatin [Yamada et al. (1998) Plant Cell Physiol. 39: 106, Yamada et al. (2000) Plant Cell Physiol. 41: 185]. Here, we show the mechanism of the SDS-dependent activation of the trimeric (P-I) complex and stabilization of the activated protease by its specific substrates. The cystatin-free cysteine protease isolated by preparative SDS-PAGE was still specifically activated by SDS, and its profile of SDS-dependency was exactly the same as that of the trimeric (P-I) complex. It is, therefore, evident that an SDS dependent conformational change of the protease itself, rather than the release of cystatin from the complex, is crucial for the activation. Pre-treatment analysis with SDS revealed that SDS was required for the initiation of the activation of the trimeric (P-I) complex. Furthermore, we found that once the protease was activated, if there was no substrate, it was rapidly inactivated under optimum conditions of proteolysis, and showed that such inactivation was not due to autolysis of the protease. In contrast, addition of specific substrates prevented the inactivation, and thus we presumed that the activity of the cysteine protease is regulated by both activation by conformational change and rapid inactivation after consumption of substrates. PMID- 11479378 TI - Involvement of calcium ion in the stimulated shoot elongation of arrowhead tubers under anaerobic conditions. AB - Shoot elongation of arrowhead (Sagittaria pygmaea Miq.) tubers was stimulated in anaerobic conditions. The anaerobic elongation was attributed to stimulation of cell elongation in the middle of the shoots. The anaerobic elongation of the shoots was severely inhibited by ethylene glycol bis(beta-aminoethylether) N,N,N',N'-tetraacetic acid (EGTA). The EGTA inhibition was completely nullified by exogenous CaCl2, which acts as an enhancer of anaerobic elongation. Moreover, calcium channel blockers, verapamil, diltiazem and LaCl3, inhibited the anaerobic elongation enhanced by CaCl2. These results showed that calcium plays an important role in stimulating the elongation in anaerobic conditions. Incorporation of 45Ca into the shoot tissues was measured to determine the involvement of calcium uptake in anaerobic elongation. Incorporation of 45Ca into the cell sap, which was collected from frozen and thawed shoots after thorough washing with LaCl3, was significantly stimulated in anaerobic conditions. Verapamil and diltiazem prevented the stimulation of 45Ca incorporation in anaerobic conditions. These results suggest that calcium uptake from the medium serves to enhance shoot elongation of arrowhead tubers under anaerobic conditions. PMID- 11479379 TI - Fate of nascent microtubules organized at the M/G1 interface, as visualized by synchronized tobacco BY-2 cells stably expressing GFP-tubulin: time-sequence observations of the reorganization of cortical microtubules in living plant cells. AB - Transgenic BY-2 cells stably expressing a GFP (green fluorescent protein)-tubulin fusion protein (BY-GT16) were subcultured in a modified Linsmaier and Skoog medium. The BY-GT16 cells could be synchronized by aphidicolin and the dynamics of their microtubules (MTs) were monitored by the confocal laser scanning microscopy (CLSM). We have succeeded in investigating the mode of reorganization of cortical MTs at the M/G1 interface. The cortical MTs were initially organized in the perinuclear regions and then they elongated to reach the cell cortex, forming the bright spots there. Subsequently, the first cortical MTs rapidly elongated from the spots and they were oriented parallel to the long axis towards the distal end of the cells. Around the time when the tips of the parallel MTs reached the distal end, the formation of transverse cortical MTs followed in the cortex near the division site, as we had previously suggested [Hasezawa and Nagata (1991) Bot. Acta 104: 206, Nagata et al. (1994) Planta 193: 567]. It was confirmed in independent observations that the appearance of the parallel MTs was followed by the appearance of the transverse MTs in each cell. We found that the transverse MTs spread through the whole cell cortex within about 20-30 min, while the parallel MTs disappeared. The significance of these observations on the mode of cortical MT organization is discussed. PMID- 11479380 TI - Isolation and characterization of oxygen-evolving thylakoid membranes and photosystem II particles from a glaucocystophyte, Cyanophora paradoxa. AB - Conditions for preparing oxygen-evolving thylakoid membranes and PSII complexes, and those for observing the PSII activity were investigated in a glaucocystophyte, Cyanophora paradoxa. The active thylakoid membranes were isolated either with a medium containing glycerol or with that containing high concentrations of sucrose, phosphate, and citrate. Active PSII particles were solubilized by octyl-beta-D-glucoside from thylakoid membranes and were separated by sucrose density gradient centrifugation. The thylakoid membranes and PSII particles showed an oxygen-evolving activity only in high-ionic-strength media. The extrinsic 33 kDa protein (PsbO) and the cytochrome c(550) (PsbV) were found to be present in the PSII particles as in cyanobacteria or red algae, but no 12 kDa protein (PsbU) was detected. The PsbO protein was classified as a land-plant type by its N-terminal amino acid sequence. PMID- 11479381 TI - Purification and characterization of Delta(1)-pyrroline-5-carboxylate reductase isoenzymes, indicating differential distribution in spinach (Spinacia oleracea L.) leaves. AB - Delta(1)-Pyrroline-5-carboxylate reductase (P5CR) (EC 1.5.1.2. L-proline: NAD(P) 5-oxidoreductase), the second enzyme in the proline biosynthetic pathway, was purified from spinach (Spinacia oleracea L.) leaves. Following ammonium sulfate fractionation, purification was performed by several chromatographic methods: Blue Cellulofine, DEAE-TOYOPEARL, Sephacryl S-300 HR, and POROS QE/M. Two isoenzymes resolved by anion exchange chromatography were designated P5CR-1 and P5CR-2. Only P5CR-2 was purified from the intact chloroplasts, indicating differential distribution of the isoenzymes. P5CR isoenzymes, P5CR-1 and P5CR-2, are a homopolymer with an apparent molecular mass of 310 kDa, consisting of 10 to 12 subunits of about 28.5 kDa. P5CR-1 and P5CR-2 showed K(m) values of 9 and 19 microM for NADPH and values of 0.122 and 0.162 mM for Delta(1)-pyrroline-5 carboxylate (P5C), respectively. We decided partial amino acid sequences of P5CR 1 which showed the 70 to 80% homology to the deduced amino acid sequences of several plant P5CR cDNAs. Both isoenzymes had much lower affinity for NADH than for NADPH and were inhibited by free ATP and Mg(2+) ion. The inhibition was partially mitigated when ATP and Mg(2+) were added simultaneously to the reaction mixture. Cations at high concentration were inhibitory to P5CR activity. Interestingly, P5CR-2 was more stable to heat treatment at 40 degrees C than P5CR 1. PMID- 11479382 TI - The AHK4 gene involved in the cytokinin-signaling pathway as a direct receptor molecule in Arabidopsis thaliana. AB - We previously identified a set of structurally related genes, AHK2, 3 and 4, each encoding a sensor histidine kinase in Arabidopsis thaliana. To determine the relevant biological functions, we identified a loss-of-function mutation of the AHK4 gene. The mutant exhibited the cytokinin-resistant phenotype not only in inhibition of root growth by cytokinin but also in greening and shoot induction of calli. Moreover, AHK4 expressed in budding yeast showed histidine kinase activity in a manner dependent on the presence of cytokinin. These results strongly suggested that AHK4 is involved in the cytokinin-signaling pathway, as a direct receptor molecule, in Arabidopsis. PMID- 11479383 TI - Myxoxanthophyll in Synechocystis sp. PCC 6803 is myxol 2'-dimethyl-fucoside, (3R,2'S)-myxol 2'-(2,4-di-O-methyl-alpha-L-fucoside), not rhamnoside. AB - We identified the molecular structures of the carotenoids in Synechocystis sp. PCC 6803. Myxoxanthophyll in this cyanobacterium was myxol 2'-dimethyl-fucoside, (3R,2'S)-myxol 2'-(2,4-di-O-methyl-alpha-L-fucoside). The sugar moiety of the pigment was not rhamnose but dimethylated fucose, which has not been reported in carotenoid glycosides. The other carotenoids were beta-carotene, (3R,3'R) zeaxanthin, echinenone, (3'R)-3'-hydroxyechinenone and deoxymyxol 2'-dimethyl fucoside, (2'S)-deoxymyxol 2'-(2,4-di-O-methyl-alpha-L-fucoside). Generally, the group of polar carotenoids in cyanobacteria is referred to as myxoxanthophyll, and the structure is considered to be myxol 2'-rhamnoside. Since the name myxoxanthophyll can not specify the sugar moiety and the identification of the sugar moiety is unfeasible in many cyanobacteria, we propose the following naming convention: when the sugar moiety is unknown, the name is myxol glycoside, when known, as in the case of rhamnose and alpha-L-fucose, they should be named myxol 2'-rhamnoside and myxol 2'-alpha-L-fucoside, respectively. PMID- 11479384 TI - Azospirillum spp. metabolize [17,17-2H2]gibberellin A20 to [17,17-2H2]gibberellin A1 in vivo in dy rice mutant seedlings. AB - Azospirillum spp. are endophytic bacteria with beneficial effects on cereals- effects partially attributed to gibberellin production by the microorganisms. Azospirillum lipoferum and Azospirillum brasilense inoculated to rice dy mutant reversed dwarfism in seedlings incubated with [17,17-2H2]GA20 with formation of [17,17-2H2]GA1, showing the in vivo capacity to perform the 3beta-hydroxylation. When prohexadione-Ca, an inhibitor of late steps in gibberellin biosynthesis, was added to the culture medium, no complementation was observed and no [17,17 2H2]GA1 was produced. The latter suggests that the bacterial operating enzyme may be a 2-oxoglutarate-dependent dioxygenase, similar to those of plants. PMID- 11479385 TI - RNA editing of the ribosomal protein S13 transcripts in magnolia and sunflower mitochondria. AB - To obtain information on ribosomal protein gene content and RNA editing in plant mitochondria, we investigated the conservation of the gene for ribosomal protein S13 (rps13) in the mitochondrial genomes of Magnolia spp. and Helianthus annuus. A complete rps13 open reading frame was identified in the mitochondrial genome of Magnolia and Helianthus. Comparison of genomic and cDNA sequences revealed that transcripts from the mitochondrial rps13 locus in both Magnolia and Helianthus undergo RNA editing suggesting the functionality of the related gene. Six and four editing events in the rps13 transcript population of Magnolia and Helianthus mitochondria, respectively, cause critical changes of codons and, consequently, induce 5 and 3.4% amino acid modifications in the respective genomically-encoded S13 polypeptides. Both editing pattern and efficiency of RNA editing differ in the reverse transcription (RT)-PCR-derived cDNA populations from the rps13 locus from the analyzed plant species with only the Magnolia rps13 being fully edited. PMID- 11479386 TI - In vivo detection of tobacco mosaic virus-induced local and systemic oxidative burst by electron paramagnetic resonance spectroscopy. AB - This is the first demonstration that tobacco mosaic virus-induced oxidative stress in a necrotic host plant is signalled by an elevated level of monodehydroascorbate (MDA) radicals detected by electron paramagnetic resonance spectroscopy. Furthermore, systemic acquired resistance induced in remote leaves of Xanthi-nc tobacco is also associated with stimulated MDA signals indicative of a microoxidative burst. PMID- 11479387 TI - The role of papillae during the infection process in the Gunnera-Nostoc symbiosis. AB - Gunnera manicata L. glands consist of up to nine separate papillae. Surgical removal of papillae showed that more than two papillae were needed for successful infection with Nostoc. Infection occurs only in the enclosed space between adjacent papillae. Dividing Gunnera cells in the enclosed space are the sites of infection. PMID- 11479388 TI - Development of UV defense mechanisms during growth of spinach seedlings. AB - Changes in UV defense mechanisms were studied during the growth periods of spinach seedlings grown under the white light, which did not contain UV-B light. DNA photolyase activity in the photorepair of cyclobutane pyrimidine dimers in spinach seedlings was high in the early growth phase (cell division phase) and declined thereafter, whereas UV-absorbing substances accumulated throughout the growth period acted as a major UV-defense mechanism in the cell expansion phase. PMID- 11479389 TI - Paroxysmal hemicranias. PMID- 11479390 TI - Neurobiology and clinical pharmacology of obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is a brain disorder with recognizable periods of onset, course, familial occurrence, epidemiology, phenomenology, and treatment response. Several manifestations of pathophysiology are beginning to be defined, although they may represent intermediate pathophysiology rather than primary etiology. Positron emission tomography studies have consistently identified hypermetabolism in the orbitofrontal cortex, caudate nucleus, and, sometimes, anterior cingulate cortex. Neuropsychologic testing frequently identifies abnormalities in visuospatial function. Abnormal levels of cerebrospinal fluid neurotransmitters and neuromodulators are identifiable in untreated patients with OCD and return toward normal levels after effective treatment. The most consistent pathophysiologic finding in OCD points toward an abnormality in serotonin neurotransmission. Therapeutic response to selective serotonin reuptake inhibitors and the absence of improvement with norepinephrine reuptake inhibitors and dopamine antagonists argue strongly for a role of serotonin in the pathophysiology and treatment of OCD. Despite this clear indication from treatment trials, probes and manipulations of the serotonin system and its specific receptors have not provided a useful understanding of specific abnormalities. Clomipramine or potent selective serotonin reuptake inhibitors are the pharmacotherapy of choice for OCD, with a more limited role reserved for monoamine oxidase inhibitors. If one selective serotonin reuptake inhibitor is ineffective, others may be beneficial, in addition to the different proserotonergic and nonserotonergic augmentation strategies that could be useful in treatment of resistant OCD patients. Nondrug therapies are also important in OCD: behavioral therapy is frequently helpful and neurosurgery is sometimes helpful when other treatments fail. PMID- 11479391 TI - Methylphenidate increases the motor effects of L-Dopa in Parkinson's disease: a pilot study. AB - We determined whether methylphenidate, a dopamine transporter blocker, modifies motor, cognitive, or affective responses to L-Dopa in Parkinson's disease (PD). Five patients who reported benefit from L-Dopa/carbidopa and motor fluctuations were admitted and withdrawn from their usual antiparkinsonian medications. On 3 consecutive days in a randomized double-blinded fashion, they took 0.2 mg/kg oral methylphenidate or placebo followed 30 minutes later by a 1-hour intravenous L Dopa (2 mg/kg per h) or placebo infusion. Vital signs, tapping, walking, dyskinesias, mood, anxiety, concentration, and arousal were monitored every 30 minutes. Cognitive testing was performed before and following the infusion. Methylphenidate combined with L-Dopa led to greater peak right-hand tapping speed than either alone. Dyskinesia severity increased most when methylphenidate and L Dopa were co-administered. There were no differences between conditions on the Stroop test, digit ordering, simple reaction time, or covert orienting of attention validity effect. Methylphenidate alone led to improvement in choice reaction time. Change in self-assessed analogue ratings of mood, anxiety, arousal, or concentration did not differ between conditions. Methylphenidate increased the motor effects of L-Dopa with minimal effects on cognitive or affective functions, suggesting a physiologic role for the dopamine transporter in patients with PD with motor fluctuations. PMID- 11479392 TI - Comparison of two dosages of tolcapone added to levodopa in nonfluctuating patients with PD. AB - The efficacy and safety of two dosages of tolcapone were compared in a 12-week crossover trial involving 118 nonfluctuating patients with PD on a stable dose of levodopa (L-Dopa). At trial onset, all patients received open-label tolcapone 100 mg three times daily for 4 weeks. At week 4, 116 eligible patients entered an 8 week double-blind treatment period and were randomized to receive tolcapone three times daily at either 100 mg (group 1; n = 58) or 200 mg (group 2; n = 58) until week 8, followed by the alternative tolcapone dosage until week 12. Ratings included Unified Parkinson's Disease Rating Scale (UPDRS), Schwab & England, and patient diaries, assessed at baseline and at 4, 8, and 12 weeks. At week 4, the investigator's global assessment (IGA) of efficacy showed improvement in 76% of patients. The mean total daily L-Dopa dose and mean UPDRS scores for subscales II and III decreased significantly (p < 0.001). During the double-blind treatment period, IGA showed improvements at either or both dosages in 61% of patients; further changes in other efficacy variables were minimal and were similar with both tolcapone dosages. The most frequent adverse events were dopaminergic (nausea and dyskinesia); the most frequent nondopaminergic adverse event was diarrhea. The incidence of adverse events during double-blind treatment was slightly higher with tolcapone 200 mg three times daily (33%) than with tolcapone 100 mg three times daily (24%). The authors conclude that tolcapone dosages of 100 mg three times daily and 200 mg three times daily are well tolerated and equally effective in improving function in L-Dopa-treated nonfluctuating patients with PD. PMID- 11479393 TI - SSRIs do not worsen Parkinson's disease: evidence from an open-label, prospective study. AB - Selective serotonin reuptake inhibitors (SSRIs) have been reported to be useful in the treatment of depression in patients with Parkinson's disease (PD). However, a few reports have suggested that SSRIs may worsen parkinsonian motor symptomatology and extrapyramidal side effects have been reported in depressed patients treated with SSRIs. So far, no prospective trial comparing the effects of different SSRIs in depressed patients with PD has been performed. The aim of the present study was to assess the effects of four SSRIs (citalopram, fluoxetine, fluvoxamine, and sertraline) on motor performance and their efficacy on depression in a group of patients with PD. Sixty-two consecutive nondemented, nonfluctuating, depressed patients with PD were included in four treatment groups (15 patiens received citalopram, 16 fluoxetine, 16 fluvoxamine, and 15 sertraline). The evaluation of extrapyramidal and depressive symptomatology was performed with use of the Unified Parkinson's Disease Rating Scale (UPDRS), Beck Depression Inventory, and Hamilton Depression Rating Scale at baseline and after 1, 3, and 6 months. Fifty-two patients completed the study. UPDRS scores were not significantly modified by the add-on therapy with each of the SSRIs studied. A significant improvement in depressive symptoms from baseline to the end of the trial was obtained with all SSRIs (Beck and Hamilton scores improving; p < 0.05 according to an analysis of variance). Our findings suggest that SSRIs do not significantly worsen extrapyramidal symptomatology and may ameliorate depression in patients with PD. PMID- 11479394 TI - Response of catatonia to risperidone: two case reports. AB - The present study describes two patients, both of Yemenite origin, with catatonic schizophrenia who responded to treatment with risperidone. One had a long history of psychiatric disorder, whereas the other was a first-episode, drug-naive patient. Our observation agrees with previous reports on the use of risperidone and other novel neuroleptic agents in the treatment of catatonia of different etiologies. PMID- 11479395 TI - Amelioration of frozen gait by tandospirone, a serotonin 1A agonist, in a patient with pure akinesia developing resistance to L-threo-3,4-dihydroxyphenylserine. AB - A 71-year-old woman presented with severe akinesia, frozen gait, and compromised postural reflexes, without rigidity, tremor, or vertical gaze disturbance. With a working diagnosis of pure akinesia, we administered amantadine (150 mg/d) and L threo-3,4-dihydroxyphenylserine (DOPS) (600 mg/d), which alleviated her symptoms. When frozen gait recurred 2 months later, we increased the dose of L-threo-DOPS to 900 mg/d and added levodopa (300 mg/d) combined with carbidopa, but this failed to improve the patient's symptoms. We then combined administration of tandospirone, a serotonin (5-HT) 1A agonist with L-threo-DOPS (600 mg/d), resulting in marked clinical improvement. Tandospirone is reported to activate noradrenergic neurons via the 5-HT 1A receptor, which could account for such striking improvement in a patient previously responsive to the noradrenergic precursor L-threo-DOPS given alone. PMID- 11479396 TI - Influence of the dopamine agonist alpha-dihydroergocryptine on the pharmacokinetics of levodopa in patients with Parkinson's disease. AB - This study investigated whether chronic coadministration of alpha dihydroergocryptine (DHEC) altered the plasma pharmacokinetics of individualized treatments with levodopa in 12 patients with Parkinson's disease. Steady-state pharmacokinetics of plasma levodopa (L-Dopa) under combined treatment were compared with those under treatment with L-Dopa alone. There was no evidence of increased exposure to L-Dopa caused by concomitant treatment with DHEC. In contrast, additional treatment with DHEC reduced the overall exposure to L-Dopa (17.5% reduction in area under the curve; 95% CI: 23%-6%). This effect was small but statistically significant for the area under the plasma concentration-time curve, whereas tmax (time of maximum plasma concentration) and peak-to-trough fluctuation were not affected. Cmax (maximum plasma concentration), on average, was reduced to a similar extent (-14.5%; 95% CI: 38% to -17%), albeit not significantly. The magnitude of the interaction does not suggest changing the current clinical practice of up-titrating DHEC and subsequently adapting L-Dopa to the individual needs of patients. PMID- 11479397 TI - Hypothermia and phenytoin toxicity: a case report. AB - Phenytoin toxicity continues to be a common medical problem. It can present with subtle and atypical symptoms. Herein the authors report what is, to their knowledge, the first case in the literature of phenytoin toxicity that manifested solely in the form of hypothermia. PMID- 11479398 TI - Amoxapine shows an antipsychotic effect but worsens motor function in patients with Parkinson's disease and psychosis. AB - Amoxapine, a dibenzoxazepine antidepressant, has been suggested to have atypical antipsychotic properties. We tested it to control psychosis in three patients with Parkinson's disease (PD). Two patients had significant improvement in hallucinations, whereas the third could not tolerate the drug for a sufficient period. All three patients experienced a decline in motor function; two also had concomitant reduction in dyskinesias. Therefore, although we found some support for amoxapine having antipsychotic properties, this drug seems to carry a risk of worsening motor function in patients with PD. PMID- 11479399 TI - Gabapentin withdrawal syndrome. AB - Gabapentin (GBP) has gained wide acceptance in the treatment of pain, migraine, bipolar illness, and epilepsy. It has a relatively benign side effect profile, lacks significant drug interactions, is not liver metabolized, and is renally excreted. Herein three cases are presented that demonstrate withdrawal symptoms after abrupt discontinuation of GBP. Clinicians are encouraged to taper GBP dosage, especially when patients have taken high doses, and to warn patients of possible adverse effects of abruptly discontinuing GBP themselves. PMID- 11479400 TI - Bacterial [correction of baterial] translocation in humans. AB - Bacterial translocation is a phenomenon in which live bacteria cross the intestinal barrier. The definition may be broadened to include transmural passage of bacterial cell wall components such as lipopolysaccharide and peptidoglycan polysaccharide. After translocation, bacteria or their products reach the mesenteric lymph nodes. From there, it is possible that enteric bacteria, their cell wall components, or both may disseminate throughout the body, causing sepsis, shock, multisystem organ dysfunction, or death of the host. Bacterial translocation and its complications have been shown clearly to occur in animal models, but its existence and importance in humans has been difficult to ascertain. The purpose of this review is to evaluate the data from studies in humans on the occurrence of bacterial translocation and, more importantly, to evaluate its role as a cause of death in humans. Studies from trauma and intensive care centers often imply that bacterial translocation is a major contributor to sepsis, shock, and multisystem organ failure in humans. However, the data reviewed herein do not support that view clearly. Carefully designed studies are needed to determine the relevance of bacterial translocation in human disease. PMID- 11479401 TI - Fecal calprotectin: a significant step in the noninvasive assessment of intestinal inflammation. PMID- 11479402 TI - Fecal calprotectin: validation as a noninvasive measure of bowel inflammation in childhood inflammatory bowel disease. AB - BACKGROUND: Calprotectin is an abundant neutrophil protein, which is extremely stable in feces. This study aimed to validate fecal calprotectin as a marker of bowel inflammation against invasive measures in children with inflammatory bowel disease (IBD), including colitis and small bowel Crohn disease. METHODS: Fecal calprotectin was measured using a simple enzyme-linked immunosorbent assay in 36 spot stool samples from 22 children before colonoscopy and from 14 children before technetium-99 (99Tc) scanning. Using standard scoring systems, the severity of inflammation was assessed macroscopically and histologically at six standard sites in those who underwent colonoscopy and also at six standard sites in those who underwent 99Tc scanning. The subscores from each site were summated to give combined severity and extent scores for macroscopic and for histologic inflammation in the group undergoing colonoscopy and total inflammation in the group undergoing 99Tc scanning. RESULTS: In the 22 children who underwent colonoscopy, median fecal calprotectin was 4.9 mg/L (0.1-272.5 mg/L) (range). Disease groups included six normal cases, nine ulcerative colitis cases, two isolated Crohn colitis cases, two indeterminate colitis cases, and three allergic colitis cases. Fecal calprotectin correlated closely with colonic macroscopic inflammation (r = 0.75, P < 0.001) and histologic inflammation (r = 0.85, P < 0.001). Of the 14 children undergoing 99Tc scanning, 10 had Crohn disease, 3 had ulcerative colitis, and 1 had allergic colitis. Median fecal calprotectin was 9.1 mg/L (0.3-141.7 mg/L), and this correlated closely with the 99Tc scanning score (r = 0.80, P = 0.001). CONCLUSION: Fecal calprotectin correlates closely with the best invasive measures of colonic and small bowel inflammation in childhood inflammatory bowel disease. As a sensitive objective measure of bowel inflammation that is risk-free and noninvasive, fecal calprotectin lends itself particularly to the monitoring of and assessment of therapeutic interventions in children with inflammatory bowel disease. PMID- 11479403 TI - Constipation in very-low-birth-weight children at 10 to 14 years of age. AB - BACKGROUND: The authors documented that very-low-birth-weight children (less than 750-g) when compared with 750-to 1,499-g birth-weight and normal-birth-weight matched controls have higher than normal rates of neurosensory and cognitive impairments at school age. In this population, the authors found a higher rate of constipation than expected among 19 of 59 (32%) less-than-750-g-birth-weight children. The authors therefore wanted to evaluate in greater detail the problem of constipation and associated toileting problems in this population. METHODS: Nineteen less-than-750-g-birth-weight children with constipation were matched to the next less than 750-g-birth-weight child of comparable age, sex, and race who did not have constipation. A telephone questionnaire was administered to 15 of 19 parents of the less-than-750-g-birth-weight children who had constipation. RESULTS: The 15 children with constipation had significantly higher rates of neurosensory impairment (7 [47%] vs. 1 [7%]) and lower mean WISC III IQ scores (72 +/- 22 vs. 89 +/- 13) when compared with birth-weight-matched controls without constipation (P < 0.05). The constipated children also had significantly higher rates of associated toileting problems, including encopresis (8[53%] vs. 1 [7%])and stool withholding (9 [60%] vs. 2 [13%]; P < 0.05). CONCLUSION: The higher incidence of constipation among less-than-750-g-birth-weight children is associated with neurodevelopment impairment. Awareness of this problem and preventive intervention to ensure regular stools could possibly decrease the high rate of constipation and associated toileting problems. PMID- 11479404 TI - Pancreatic elastase 1 in feces of preterm and term infants. AB - BACKGROUND: Determination of fecal pancreatic elastase 1 (E1) is a reliable and noninvasive test of exocrine pancreatic function. Adult reference values of greater than 200 microg E1/g feces do not seem to be applicable to early infancy because of immature pancreatic function. Because reference values for infants do not exist, the current study was aimed to define reference values for preterm and term infants up to 12 months of age. METHODS: The authors measured pancreatic E1 concentration in feces of 148 infants up to 12 months of age. Infants with known bowel or pancreatic disorders were excluded from the study. RESULTS: The authors found that 96.8% of all children had E1 concentrations greater than an adult lower limit after 2 weeks of life, independent of gestational age. Up to 48 hours after birth, none of the preterm infants had an E1 concentration of greater than 30 microg/g meconium, whereas 43% of the term infants had normal adult values. CONCLUSIONS: The adult reference value for pancreatic E1 of greater than 200 microg/g feces can be applied to infants older than 2 weeks, independent of gestational age, birth weight, and the type of nutrition. PMID- 11479405 TI - Phenotypic and genetic analysis of diarrhea-associated Escherichia coli isolated from children in the United Kingdom. AB - BACKGROUND: A hospital-based study was performed to (1) compare phenotypic and genotypic diagnostic tests for enteropathogenic Escherichia coli, enteroaggregative E. coli, and diffuse-adhering E. coli (collectively termed adherent E. coli) and (2) to assess the importance of these different classes of adherent E. coli as causes of infant diarrhea in the United Kingdom in comparison with other enteropathogens. METHODS: E. coli isolated from 1,496 infants with diarrheal disease and from 546 age-related controls were screened for enteropathogenic E. coli, enteroaggregative E. coli, and diffuse-adhering E. coli using HEp-2 cell adherence assays and DNA probes. RESULTS: Marked discrepancies between the phenotype and genotype of isolates indicate significant heterogeneity among enteroaggregative E. coli and diffuse-adhering E. coli strains. Depending on the assay used, adherent E. coli were isolated as the only putative pathogen in 23% to 27% of diarrhea cases, a significantly higher incidence than in the control group. Individually, enteroaggregative E. coli (8.5-8.6% of cases) and diffuse-adhering E. coli (10.4-11.3% of cases), but not enteropathogenic E. coli (4.5-7.5% of cases), were significantly associated with diarrhea. CONCLUSIONS: These studies indicate that adherent E. coli may be an important cause of diarrhea in infants in the United Kingdom; they also emphasize the need for more specific virulence-based tests for these putative classes of "diarrheagenic" (diarrhea causing) E. coli. PMID- 11479406 TI - Cisapride plasma levels and corrected QT interval in infants undergoing routine polysomnography. AB - BACKGROUND: Reported QTc prolongation associated with cardiac arrhythmia in a small number of children undergoing cisapride therapy and lack of pharmacokinetic correlation provided the impetus for this prospective study. The authors evaluated the relation between cisapride plasma concentrations, the electrocardiographic QT interval, and cardiac rhythm in infants undergoing routine 8-hour polysomnography. METHODS: A total of 211 infants were enrolled: 84 (17 born prematurely) undergoing cisapride therapy for at least 4 days for suspected gastroesophageal reflux and 127 controls (10 born prematurely), aged between 1 week and 13.5 months. Infants underwent continuous bipolar limb lead I recording during routine 8-hour polysomnography. QT intervals and heart rate were measured at hourly intervals. The morning after polysomnography, 12-lead electrocardiography was performed (1 hour after cisapride administration). Cisapride plasma concentrations were determined immediately before and 1 to 2 hours after administration. Serum electrolyte concentrations were measured. RESULTS: The administered cisapride dose ranged from 0.35 to 1.55 (mean, 0.81, median 0.79) mg. kg-1. d-1. Cisapride plasma concentrations were significantly higher in infants younger than 3 months of age. Cisapride-treated infants younger than 3 months of age had longer QTc intervals compared with age-matched controls. Heart rate was similar for cisapride-treated and control infants. No arrhythmia or atrioventricular conduction abnormalities were observed. CONCLUSIONS: At comparable doses of cisapride and comparable plasma concentrations, the QTc was significantly higher in infants younger than 3 months of age. This confirms age dependent cisapride pharmacokinetics in the first 10 to 12 weeks strongly correlated with changes in body weight and may also suggest an altered ability of infants younger than 3 months of age to metabolize cisapride. The clinical significance and risk of the increased QTc interval is unclear. Cisapride should be judiciously prescribed in infants younger than the age of 3 months and electrocardiography should be performed before and during therapy. PMID- 11479407 TI - Autonomic abnormalities in children with functional abdominal pain: coincidence or etiology? AB - BACKGROUND: There is increasing evidence that autonomic neuropathies may adversely affect gastrointestinal motility by involving the extrinsic nerves of the gut. The authors' hypothesize that functional abdominal pain in children is associated with generalized autonomic dysfunction. METHODS: The authors performed detailed autonomic testing in eight patients with functional abdominal pain, including deep breathing, Valsalva, tilting (to assess parasympathetic and sympathetic adrenergic function), and axon-reflex function and thermoregulatory sweat testing to assess sympathetic cholinergic function. Patients also completed a questionnaire regarding other autonomic symptoms. RESULTS: Results of autonomic testing were abnormal in seven patients. Parasympathetic function was normal in all, and the abnormalities were restricted to sympathetic cardiac, vasomotor, and sudomotor function. Abnormal results of axon-reflex testing in six were consistent with peripheral nervous system dysfunction. Five had decreased sweating over the abdomen, determined by thermoregulatory sweat testing. Five eight had nongastrointestinal autonomic symptoms, primarily palpitations and flushing. CONCLUSIONS: Functional abdominal pain in the current patients is associated with generalized dysfunction of the autonomic nervous system. This dysfunction can be peripheral or central in different individuals but seems to be restricted to the sympathetic branch. The known function of the sympathetic nervous system as the motility "brake" suggests that pain could be a manifestation of unmodulated peristalsis, resulting in abdominal cramps. PMID- 11479408 TI - Laparoscopic full-thickness intestinal biopsies in children. AB - BACKGROUND: Laparoscopy may reduce postoperative pain and hospital stay, compared with laparotomy. The use of laparoscopic surgery to obtain full-thickness intestinal biopsies in children has not been previously reported. METHODS: Eleven children aged 1.6 to 19 years (median, 4.5 years) underwent laparoscopic full thickness biopsy of the stomach, small bowel, colon, or a combination thereof. Each procedure used one 12-mm and two 5-mm ports. RESULTS: Eight children with obstructive symptoms after a pull-through for Hirschsprung disease underwent multiple colon and small bowel biopsies (range, 3-6; median, 5); intestinal neuronal dysplasia was found in two. Two patients with cystic fibrosis had diffuse colonic narrowing; a diagnosis of enzyme-induced fibrosing colonopathy was made in one and nonspecific inflammation was found in the other. One child had a thickened stomach, and a gastroscopic-directed full-thickness biopsy revealed plasmacytoma. Nine of the 11 patients had a previous laparotomy, and ports were placed through preexisting scars. Median hospital stay was 2 days. No patient required more than 24 hours of narcotics. There were no leaks, and no other morbidity or mortality occurred. None of the patients required conversion to an open procedure. Biopsy results significantly affected treatment for each patient. CONCLUSIONS: Laparoscopic full-thickness intestinal biopsy is safe and effective for a variety of gastrointestinal problems in children. This technique is associated with a short hospital stay, minimal pain, and a very low risk of complications and can be performed even in patients who have had a previous laparotomy. PMID- 11479409 TI - Familial and community environmental risk factors for Helicobacter pylori infection in children and adolescents. AB - BACKGROUND: The aim of the study was to identify familial and community environmental risk factors associated with Helicobacter pylori infection in a pediatric population. METHODS: Children requiring diagnostic upper endoscopy were included in the study during a 2-year period. During endoscopy, five gastric biopsies were performed for the histologic or bacteriologic diagnosis, or both, of H. pylori infection. Epidemiologic data collected by a questionnaire were analyzed using the chi-square test or Fisher test and stepwise logistic regression. RESULTS: The authors included 436 patients (242 boys), aged 2 days to 17.9 years (median, 2.7 years). H. pylori prevalence was 7.3%. Univariate analysis found H. pylori was more common in older patients (P < 0.00001), in children who had at least one parent born in a developing country (P < 0.02) or with a low socioeconomic status (P < 0.02), and in those living in crowded conditions (P < 0.02). Children whose mother worked at home were more frequently infected than children whose mother worked outside the home (P < 0.02). Attendance at nursery or school before the age of 6 years was not associated with infection. Logistic regression showed a strong association with H. pylori only for age and number of persons at home. CONCLUSIONS: The source of H. pylori is intrafamilial rather than from a community, such as nursery and school attended at a young age. The number of persons in the home influences the infection status of children but not by the presence of the mother in home. These data suggest that H. pylori infection transmission occurs from siblings or the father rather than from mother. PMID- 11479410 TI - Thiamine, riboflavin, pyridoxine, and vitamin C status in premature infants receiving parenteral and enteral nutrition. AB - BACKGROUND: There is a paucity of data about water soluble vitamin status in low birthweight infants. Therefore, the authors' objective was to assess current feeding protocols. METHODS: The authors measured serum concentrations for riboflavin, pyridoxine, and vitamin C and functional assays for thiamine and riboflavin longitudinally in 16 premature infants (birthweight, 1,336 +/- 351 g; gestational age, 30 +/- 2.5 weeks) before receiving nutrition (time 1, 2 +/- 1 days), during supplemental or parenteral nutrition (time 2, 16 +/- 10 days) and while receiving full oral feedings (time 3, 32 +/- 15 days). In plasma, vitamin C was measured colorimetrically, and riboflavin and pyridoxine were measured using high-performance liquid chromatography. The erythrocyte transketolase test as a functional evaluation of thiamine and the erythrocyte glutathione reductase test for riboflavin were measured colorimetrically. RESULTS: At time 1, nutrient intake of vitamins were negligible because infants were receiving intravenous glucose and electrolytes only. Intakes differed between time 2 and time 3 for thiamine (510 +/- 280 and 254 +/- 115 microg. kg-1. d-1, respectively), riboflavin (624 +/- 305 and 371 +/- 193 microg. kg-1. d-1, respectively), and pyridoxine (394 +/- 243 and 173 +/- 85 microg/100 kcal, respectively), but not for vitamin C (32 +/- 17 and 28 +/- 12 mg. kg-1. d-1, respectively). Blood levels at times 1, 2, and 3 were for thiamine (4.9 +/- 2.7%, 3.3 +/- 6.6%, and 4.1 +/- 9% erythrocyte transketolase test, respectively), riboflavin (0.91 +/- 0.31, 0.7 +/- 0.3, 0.91 +/- 0.18 erythrocyte glutathione reductase test, respectively), riboflavin (19.5 +/- 17, 23.3 +/- 8.6, 17.6 +/- 10 ng/mL, respectively), pyridoxine (32 +/- 25, 40 +/- 16, 37 +/- 26 ng/mL, respectively), and vitamin C (5.2 +/- 3, 5 +/- 2.2, 10 +/- 5 microg/mL, respectively) and did not differ at those times. CONCLUSIONS: Current intakes of these vitamins, except for possibly vitamin C, during parenteral and enteral nutrition seem to result in adequate plasma concentrations and normal functional indices. PMID- 11479411 TI - Influence of oral inoculation with plasmid-free human Escherichia coli on the frequency of diarrhea during the first year of life in human newborns. AB - BACKGROUND: This study was carried out to determine whether early inoculation of the plasmid-free human Escherichia coli into human newborns would reduce the frequency of acute diarrhea during a 1-year period. The plasmid-free E. coli strain isolated from the fecal microbiota of a healthy adult was nontoxigenic in vivo and in vitro and sensitive to all usual antibiotics. METHODS: In the experimental group, 51 healthy newborns were inoculated orally with 106 viable cells of the bacteria within 2 hours after birth. In the control group, the same number of newborns received the heat-killed bacteria. The clinical trial was double blind, and the newborns were randomly assigned to the experimental and control groups. RESULTS: Six months and 1 year after bacterial inoculation, infants in the experimental group showed a higher mean body weight (7.59 +/- 1.15 kg and 9.88 +/- 1.31 kg, respectively; P < 0.05) when compared with the control group (7.03 +/- 1.09 kg and 8.92 +/- 1.38 kg, respectively). At the end of the clinical trial, 48% (23/48) of the infants in the experimental group had shown at least one diarrhea episode during the 1-year period, as opposed to 71% (34/48) in the control group. These values were significantly different (P = 0.037), showing a 32.3% protective effect of inoculation. CONCLUSIONS: The present study shows that protection against diarrhea was obtained by oral inoculation with a single dose of plasmid-free human E. coli soon after birth. PMID- 11479412 TI - Controlled ileal release budesonide in pediatric Crohn disease: efficacy and effect on growth. AB - BACKGROUND: Although the effectiveness of controlled ileal release (CIR) budesonide in children can be extrapolated from adult studies, there are currently no data available concerning the effects of CIR budesonide therapy on linear growth. In the absence of controlled, prospective pediatric clinical trials, we reviewed the outcomes, particularly linear growth, of children and adolescents given CIR budesonide to treat active intestinal inflammation and to maintain remission. METHODS: Thirty-two children (20 males) aged 14.1 +/- 2.7 years with Crohn disease of the distal ileum with or without right colon involvement were treated for active Crohn disease (baseline Pediatric Crohn Disease Activity Index, 34 +/- 14) with 9 mg daily of CIR budesonide through the Hospital for Sick Children, University of Toronto, Inflammatory Bowel Diseases program. RESULTS: At first follow-up visit 8.7 +/- 6.0 weeks later, 19 of 32 (59%) were judged by the physician to have responded. In the subset of 22 patients who had laboratory tests repeated at the first follow-up visit, their Pediatric Crohn Disease Activity Index fell from 33 +/- 14 to 22 +/- 16 (P = 0.001). The Pediatric Crohn Disease Activity Index score fell to less than 15 (cut-off score remission) in 29%. Six prepubertal responders continued to receive 6 mg CIR budesonide for 6 to 13 months. Five of the 6 experienced only mild or no gastrointestinal symptoms and gained weight. Nevertheless, their mean height velocity was only 2.3 +/- 1.0 cm/year, and none grew at a rate of more than 4cm/year whilst receiving CIR budesonide. CONCLUSIONS: These data provide grade III evidence of modest effectiveness of CIR budesonide in children with active Crohn disease confined to the ileum with or without right colon involvement. The subnormal growth observed with continued therapy is concerning and may reflect either inadequately controlled intestinal inflammation or direct suppression of linear growth, as is observed with conventional corticosteroids. Randomized controlled pediatric trials of CIR budesonide must include parameters of linear growth as an outcome variable. PMID- 11479413 TI - Accidental caustic ingestion in children: is endoscopy always mandatory? AB - BACKGROUND: Patients who have experienced severe caustic injury to the gastrointestinal tract are at high risk of esophageal strictures. Early endoscopy is usually recommended systematically in children after caustic ingestion to assess the severity of the initial digestive lesions. The aim of this study was to determine the predictive value of clinical symptoms and ingested-substance types as markers of severe esophagogastric lesions and to define indications for endoscopy. METHODS: Ingested-product types, clinical symptoms, endoscopic data and outcome were prospectively recorded in 85 children admitted after accidental caustic ingestion. RESULTS: Forty-eight children (57%) had no symptoms; the others presented with vomiting, hematemesis, drooling, respiratory distress, and/or oropharyngeal lesions. Endoscopy showed no or minimal lesions in 63 cases (74%). None of the children developed digestive sequelae. Severe esophagogastric lesions were present in 22 cases (26%), mostly caused by lye ingestion (14 of 22) but also by strong acids (4 of 22); 9 of the 22 children (41%) developed esophageal stenosis. Vomiting, drooling, and oropharyngeal lesions did not predict severe endoscopic lesions. Hematemesis, respiratory distress, or presence of at least three of the symptoms was associated with severe lesions (positive predictive value = 1). The absence of symptoms was always associated with no or minimal lesions (negative predictive value = 1). CONCLUSIONS: In conclusion, endoscopy is not recommended for children living in developed countries who are asymptomatic after accidental caustic ingestion. PMID- 11479414 TI - Reversible intestinal pseudoobstruction associated with neural crest tumors. PMID- 11479415 TI - A gastric diverticulum containing pancreatic tissue and presenting as congenital double pylorus: case report and review of the literature. PMID- 11479416 TI - Gallbladder hydrops caused by intraluminal clot in hemobilia: an unusual complication of hepatic trauma in childhood. PMID- 11479417 TI - Granulocyte-macrophage colony-stimulating factor for treating gastrostomy tube site healing in a child with glycogen storage disease type Ib. PMID- 11479418 TI - Cancer in children with celiac disease: a survey of the European Society of Paediatric Gastroenterology, Hepatology and Nutrition. AB - BACKGROUND: In adults, the relation between celiac disease (CD) and cancer has been long recognized. In children, only four cases of CD and cancer have been described in Europe. We made a new inventory of cases with CD and cancer in children that were known by the members of the European Society for Paediatric Gastroenterology, Hepatology and Nutrition between 1989 and 1999. METHODS: Postal inquiry was made of all European Society for Paediatric Gastroenterology, Hepatology and Nutrition members mentioned on the society's web page. The members were asked if they had seen a child with CD and cancer between 1989 and 1999 and if so, to supply additional clinical data. Also, information on working place, experience, and number of celiac patients under their care was requested. RESULTS: Fifty-six percent of the members responded. Sixteen members reported 22 cases of cancer and CD in children. One case had been reported in the literature previously. The tumors that were reported originated from the brain, thyroid, larynx, liver, small bowel, adrenal, lymphoreticular system, and the musculoskeletal system. There were no differences between members reporting a case and those who did not. CONCLUSIONS: Twenty-one new cases of cancer and CD in children in Europe were found. Cancer and CD in children are underreported. A remarkable number of thyroid and small bowel cancers were found, suggesting a possible relation with CD. It is important to evaluate whether these findings are coincidental. All cases of cancer and CD in children should be reported to the literature. PMID- 11479419 TI - Melanoma induction in a hairless mouse with short-term application of dimethylbenz[a]anthracene. AB - Melanomas have been induced in hamsters and guinea pigs with short-term, low dose applications of dimethylbenz [a]anthracene (DMBA) alone. In mice, however, melanoma induction has required either croton oil or ultraviolet radiation promotion in addition to DMBA. In this study, we report the development of a malignant melanoma, with metastases, in a hairless mouse after six applications of 0.25% DMBA alone. At sacrifice, a large primary tumour with characteristics of intralesional transformation was present, along with numerous pigmented macules and papules. Metastases were present in lymph nodes and lungs. There was a marked similarity between this melanoma and its precursor lesions and those seen in an earlier, Weiser-Maple guinea pig model, which, in turn, resembled human melanoma. PMID- 11479420 TI - Optimization of intracerebral tumour protection by active-specific immunization against murine melanoma B16/G3.12. AB - Development of brain metastases despite extracerebral response to systemic immunotherapy is a common problem in melanoma patients. We have previously described a murine melanoma vaccine of interferon-gamma (IFNgamma)-treated, irradiated syngeneic B16/G3.12 and allogeneic (Cloudman) melanoma cells, plus the adjuvant DETOX, that is protective against subcutaneous (93%) or intracerebral (69%) syngeneic challenge. This study aimed to optimize this vaccine. Groups of nine or 10 mice were immunized five times in 5 weeks with: (i) complete vaccine +/- IFNgamma (VAC+, VAC-); (ii) syngeneic 2 x 106 G3.12 cells plus DETOX (Syn+D), (iii) 2 x 106 allogeneic Cloudman cells plus DETOX (Allo+D); (iv) VAC+ without DETOX (no DETOX); (v) DETOX alone (DETOX); or (vi) phosphate buffered saline (PBS). Mice were challenged subcutaneously with 104 viable G3.12 (or Cloudman cells) and after 35 days intracerebrally with 104 G3.12 cells. Expression of H-2 antigens (measured using fluorescence-activated cell sorting), splenocyte cytotoxicity (measured using 51Cr release) and median overall survival (OAS) were analysed using the log-rank test. VAC+, VAC- and G3.12 mice were equally protected from subcutaneous (s.c.) and intracerebral (i.c.) melanoma challenge (OAS 65 days for s.c., 30 days for i.c.). Protection was less (P < 0.05) in DETOX mice (48 days for s.c.), PBS mice (47 days for s.c., 21 days for i.c.) or no DETOX mice (51 days for s.c.). Allo+D mice showed s.c. (59 days) but not i.c. protection (20 days). IFNgamma incubation did not increase the effect in either the challenge cells or the vaccine cells (P > 0.05). Specific cytotoxicity was seen with G3.12 targets in VAC+ (27%) but not PBS (2%; P < 0.05) mice with equal NK (YAC-1) lysis (10% versus 7%; P< 0.05). Optimal protection against s.c./i.c. experimental murine melanoma was yielded by irradiated syngeneic cells plus DETOX. DETOX alone was not active. Upregulation of H-2 antigens with IFNgamma under these conditions does not augment protection. PMID- 11479421 TI - Growth inhibition of human malignant melanoma transfected with the human interferon-beta gene by means of cationic liposomes. AB - Among the various types of human interferons, human interferon-beta (HuIFNbeta) has the strongest anti-proliferative activity against human melanoma cell lines. Therefore, we investigated the growth inhibitory effect of a cationic liposome containing the HuIFNbeta gene on human melanoma cell lines in vitro and in vivo. After transfection with liposomes containing the HuIFN-beta gene, human melanoma cell lines produced HuIFNbeta in the culture medium at levels ranging from 67 to 3.8 IU/ml on day 6, and growth of the cells was inhibited by 71-92%. Moreover, six injections of liposomes containing the HuIFNbeta gene completely eradicated human melanoma nodules transplanted onto the backs of nude mice 40 days after the first injection. Histological analysis of the injected nodules revealed that the HuIFNbeta gene transfection induced apoptosis of the human melanoma cells. These data suggest that transfection of the HuIFNbeta gene using cationic liposomes is a promising candidate for gene therapy of human melanoma. PMID- 11479422 TI - No evidence of a role for activating CDK2 mutations in melanoma. AB - Inactivation of p16INK4a and/or activation of cyclin-dependent kinase-4 (CDK4) are strongly associated with both susceptibility and progression in melanoma. Activating CDK4 mutations prevent the binding and inhibition of CDK4 by p16INK4a. A second, more indirect role for CDK4 is in late G1, where it may sequester the inhibitors p27KIP1 or p21CIP1 away from CDK2, and in doing so upregulate the CDK2 activity necessary for cells to proceed completely through G1 into S phase. As the pivotal residues around the most predominant R24C activating CDK4 mutation are invariant between CDK2 and CDK4, we speculated that the pivotal arginine (position 22 in CDK2), or a nearby residue, may be mutated in some melanomas, resulting in the diminution of its binding and inhibition by p27KIP1 or p21CIP1. However, except for a silent polymorphism, we detected no variants within this region of the CDK2 gene in 60 melanoma cell lines. Thus, if CDK2 activity is dysregulated in melanoma it is likely to occur by a means other than mutations causing loss of direct inhibition. We also examined the expression of the CDK2 gene in melanoma cell lines, to assess its possible co-regulation with the gene for the melanocyte-lineage antigen pmel17, which maps less than 1 kb away in head to head orientation with CDK2 and may be transcribed off the same bidirectional promoter. However, expression of the genes is not co-regulated. PMID- 11479423 TI - Heterogeneity of allelic deletions within melanoma metastases. AB - During the initiation and progression of malignant melanoma, a series of different genetic events accumulate on several different chromosomes. The biological heterogeneity of tumour cells presents a major problem, preventing effective treatment of melanoma. To examine the degree of genetic heterogeneity, we searched for allelic losses (loss of heterozygosity; LOH) on chromosomes 9p, 9q, 1p and 17p, examining different areas within human melanoma metastases. All of the examined metastases were informative within at least one dissected area for at least one marker. Out of 29 areas in 11 melanoma metastases, 58% showed LOH with at least one marker. On chromosome 9p21-22, eight out of 26 informative loci (31%) showed LOH at D9S171 (three not informative), two out of 18 (11%) at IFNA (11 not informative) and seven out of 24 (29%) at D9S169 (five not informative). LOH on chromosome 9q22.3 was examined by the microsatellite marker D9S12; three out of 24 areas (12.5%) showed LOH, and five were not informative. Deletions on chromosome 1p were assessed using D1S450. Four out of 25 (16%) showed LOH; four were not informative. Deletions on chromosome 17p13 were examined with TP53; two out of 21 cases (9%) showed LOH, and eight were not informative. Our data demonstrate an impressive heterogeneity of allelic losses in the investigated chromosomal areas within the same metastatic lesion. This suggests that there is not one specific genetic alteration that accounts for melanoma progression to metastases. Rather there seem to be multiple genetic alterations accumulating even on the same chromosome, and progression from melanoma to metastases is paralleled by the accumulation of clones harbouring multiple genetic abnormalities. PMID- 11479424 TI - Loss of expression of protein kinase C beta is a common phenomenon in human malignant melanoma: a result of transformation or differentiation? AB - As with most cancers, the aetiology of human cutaneous melanoma is likely to be multifactorial and to include the accumulation of irreversible alterations in an unknown number of genes. Elucidating this molecular progression necessitates both the identification of genetic perturbations at each clinically relevant stage, and the assessment of their impact on the normal melanocyte. The observation that the epidermal melanocyte, in contrast to metastatic melanoma cells, requires activation of the protein kinase C (PKC) pathway to facilitate growth in vitro indicates that one or more isoforms (or substrates) of this large and complex family of proteins are among those that undergo alteration during the development of malignant melanoma. Consequently, a number of studies have investigated the expression of various PKC family members in both melanocyte and melanoma cell lines, without a consensus of opinion as to which isoforms are of biological significance in melanoma development and progression. The present study involved a comprehensive evaluation of the PKC profile in normal melanocytes and in 16 metastatic melanoma cell lines. The results show that the major difference in isoform expression between epidermal melanocytes and melanoma cells is the loss of PKCbeta protein expression in 90% of melanoma cell lines. Examination of PKCbeta in benign and malignant melanocytic lesions revealed that this protein is either downregulated or absent in both naevi and metastatic melanomas. We conjecture that, although the loss of PKCbeta expression is a common phenomenon in malignant melanocytes, it may be related more to a normal process of melanocytic differentiation than to malignant transformation. PMID- 11479425 TI - Reliability of PCR-based detection of occult tumour cells: lessons from real-time RT-PCR. AB - For the molecular detection of rare tumour cells in clinical samples, real-time reverse transcription-polymerase chain reaction (RT-PCR) offers two important advantages over conventional RT-PCR assays: the results are quantitative and, perhaps more importantly, it facilitates exact sensitivity controls on a per sample basis as well as exact comparison of different assay protocols. We report here on quantitative results obtained with different protocols for RNA isolation and cDNA synthesis for amplification of beta2-microglobulin transcripts using the light cycler system. Furthermore, housekeeping gene-specific PCRs were compared with PCRs specific for an artificial transcript (internal standard) detected simultaneously at a level comparable to the wild-type sequence. Artificial tyrosinase transcripts derived from a vector construct stably transfected into a human lymphoma cell line were used as a model to test the usefulness of artificial internal standards as an alternative to housekeeping genes. The highest RNA yields were obtained using a combination of phenol-chloroform extraction and the High Pure RNA Isolation Kit. Analysing beta2-microglobulin transcript-specific RT-PCRs, the highest sensitivity was obtained for cDNAs generated with Omniscript reverse transcriptase and oligo-p(dT)15 primer. Regarding patient blood samples, RT-PCRs specific for beta2-microglobulin, porphobilinogen deaminase and artificial tyrosinase transcripts provided quantitative data for all, for 18 out of 21, and for 10 out of 21 samples, respectively. Quantification of beta2-microglobulin transcripts by the light cycler system defined the protocol revealing the highest cDNA quality. Comparisons of quantitative data from RT-PCRs specific for beta2-microglobulin, porphobilinogen deaminase and artificial tyrosinase transcripts enabled us to determine a close range for crossing points within which sufficient cDNA quality can be guaranteed, even for the detection of rare transcripts. PCRs specific for the artificial internal standard are ideally suited for cDNA quality assessment on a per sample basis. PMID- 11479426 TI - Differential expression levels of Par-4 in melanoma. AB - The pro-apoptotic prostate apoptosis response-4 gene product Par-4 sensitizes prostate cells to the induction of programmed cell death. In this study we examined Par-4 expression in human melanoma cell lines and melanoma metastases. The heterogeneous expression detected prompted us to investigate the biological relevance of Par-4 in a human melanoma xenotransplantation model. Overexpression of Par-4 by transfection decreased tumour development in xenotransplanted A375-C6 melanoma cells in SCID mice and correlated to an increase in tumour cell apoptosis. These data suggest that high expression of the pro-apoptotic protein Par-4 could qualify as a prognostic marker in human melanoma. PMID- 11479427 TI - Elevated procaspase levels in human melanoma. AB - In this study procaspase expression levels were investigated by Western blotting in a panel of established melanoma cell lines, transformed melanocytic cell lines and normal primary melanocytes. Upstream caspases such as procaspase-8 that contain a death effector domain were found to be overexpressed in transformed melanocytes and melanoma cell lines compared with melanocytes. Heterogeneous levels of procaspase-8 were seen in melanoma cells, including one cell line that completely lacked procaspase-8 expression. Procaspase-10 is generally overexpressed in transformed melanocytes and melanoma cell lines. Expression of the downstream procaspases-3 and -7 was increased in melanoma cells compared with normal melanocytes. Procaspases containing caspase recruitment domains such as procaspase-2 were expressed at similar levels in nearly all the cell lines investigated. Reduced levels of procaspase-1 compared with normal melanocytes were detected in transformed melanocytes and melanoma cell lines. These data indicate that procaspase levels in general increase during the malignant transformation of melanocytic cells. PMID- 11479428 TI - Serum imbalance of cytokines in melanoma patients. AB - The cytokines interleukin (IL)6 and IL10 appear to be involved in the progression of melanoma because they are secreted by malignant cells and their serum levels are associated with poor survival and with advanced stages of the disease. Antitumour immunity is considered to be a T-cell response, mediated mainly by type 1 cytokines such as IL12 and interferon-gamma (IFNgamma). We evaluated the serum levels of cytokines involved in the host response against tumour (IL12, IFNgamma) and/or the progression of melanoma (IL6, IL10) in 45 melanoma patients with localized and metastatic disease and in 45 controls, using commercially available enzyme-linked immunosorbent assay (ELISA) kits. In the controls, IL6 and IL12 were nearly undetectable, whereas the IL10 and IFNgamma ranges were 0.5 9 pg/ml and 2-4.8 pg/ml, respectively. In the melanoma patients, pathologically high values were found in 44.4% for IL6, in 24.4% for IL10, and in 60% for IL12. Significantly higher values were found for IL6 and IL12, and lower values for IFNgamma. This study highlights a significant difference in serum cytokine profiles between controls and melanoma patients, which is mainly due to the high levels of IL6 and IL12 and the low levels of IFNgamma. PMID- 11479429 TI - The morphology, immunophenotype and distribution of paracortical dendritic leucocytes in lymph nodes regional to cutaneous melanoma. AB - Our aim was to identify and delineate alterations in the distribution and immunophenotype of the lymphocytes and paracortical dendritic leucocytes (interdigitating dendritic cells; IDCs) in lymph nodes regional to tumours. Using immunocytochemistry and computer-assisted image analysis we examined 65 lymph nodes excised from 47 patients with malignant melanoma. Twenty-nine patients had American Joint Committee on Cancer (AJCC) stage II melanoma (no tumour spread beyond the primary site) and 18 had AJCC stage III disease (metastases in the regional nodes). There were significant differences in the frequency, morphology, immunophenotype and anatomical distribution of the IDCs and in the complexity of their dendritic processes in different areas within individual lymph nodes. We conclude that morphological and phenotypical variations in IDCs correlate with differing levels of antigen presentation. Downregulation of antigen presentation in lymph nodes regional to tumours is most probably mediated by tumour products. Differences in IDC distribution and characteristics in lymph nodes from different anatomical sites must be considered in interpreting studies of nodal morphology and function. PMID- 11479430 TI - Cisplatin and carboplatin combination as second-line chemotherapy in dacarbazine resistant melanoma patients. AB - High-dose cisplatin regimens have been shown to be highly active in advanced melanoma patients but are associated with unacceptable side effects. In order to increase the platinum dose but avoid severe side effects, we treated 15 dacarbazine (DTIC)-resistant metastatic melanoma patients with a combination regimen of cisplatin (100 mg/m2) and carboplatin (200 mg/m2), two platinum analogues with a similar mode of action but a different toxicity pattern. After a mean follow-up period of 10.7 months (range 4-18 months), two patients (13.3%) achieved complete remission and two patients (13.3%) showed partial remission, giving an overall response rate of 26.4% (95% confidence interval [CI] 4.2-49%). Furthermore, three patients (20%; 95% CI 0-40.2%) experienced stable disease. The median duration of response was 7.1 months (95% CI 4.2-10.0 months), and the median overall survival was 12.5 months (95% CI 5.8-19.2 months), with eight patients still alive. The main side effects were haematological (leukopenia/thrombocytopenia World Health Organization [WHO] grade I-IV; anaemia WHO grade I-III), gastrointestinal (WHO grade I-III), neurological (WHO grade I II) and renal (WHO grade I) toxicity. Nevertheless, except in one patient, side effects did not result in discontinuation of therapy. Despite the small number of patients treated in this preliminary study, we believe that combining cisplatin and carboplatin represents a novel, active and well-tolerated therapeutic option as second-line chemotherapy in DTIC-resistant advanced melanoma patients. PMID- 11479431 TI - Functional role of melanoma inhibitory activity in regulating invasion and metastasis of malignant melanoma cells in vivo. AB - MIA (melanoma inhibitory activity) has been previously isolated from the tissue culture supernatant of melanoma cell lines as an autoregulatory activity, inhibiting thymidine incorporation. However, subsequent analyses of melanocytic tumours in vivo have correlated enhanced MIA expression with progression of melanocytic tumours, conflicting with the idea that MIA acts as a tumour suppressor. To investigate the role of MIA in vivo, we have therefore generated a panel of stably transfected B16 cell clones secreting different amounts of MIA. The capacity of these cell clones to form lung metastases in syngeneic C57Bl6 mice was strictly correlated to the level of MIA secretion, but the clones did not differ with respect to their proliferation in vitro. In summary, we suggest that MIA plays a causal role in promoting the metastasis of malignant melanomas, involving inhibition of tumour cell attachment to extracellular matrix molecules within their local milieu. PMID- 11479432 TI - Pharmacokinetics and pharmacodynamics of melphalan in isolated limb infusion for recurrent localized limb malignancy. AB - Isolated limb infusion (ILI) is an attractive, less complex alternative to isolated limb perfusion (ILP). It has a lower morbidity in treating localized recurrences and in transit metastases of the limb for tumours such as melanoma, Merkel cell tumour and Kaposi's sarcoma, allowing administration of high concentrations of cytotoxic agent to the affected limb under hypoxic conditions. Melphalan is the preferred cytotoxic agent for the treatment of melanoma by ILP or ILI. We report pharmacokinetic data from 12 patients treated by ILI for tumours of the limb in Brisbane. The kinetics of drug distribution in the limb was calculated using a two-compartment vascular model, where both tissue and infusate act as well-stirred compartments. Analysis of melphalan concentrations in the perfusate during ILI showed good agreement between the values measured and the concentrations predicted by the model. Recirculation and wash-out flow rates, tissue concentrations and the permeability surface area product (PS) were calculated. Correlations between the PS value and the drug concentrations in the perfusate and tissue were supported by the results. These data contribute to a better understanding of the distribution of melphalan during ILI in the limb, and offer the opportunity to optimize the drug regimen for patients undergoing ILI. PMID- 11479433 TI - [Total thyroidectomy in the surgical treatment of thyroid disease a retrospective clinical study]. AB - BACKGROUND: Thyroid diseases constitute a group of benign and malignant affections, among which the most represented is multinodular goitre. Nowadays, an important question regards routine total thyroidectomy as the treatment of choice for all thyroid diseases. The aim of the present study is to verify if total thyroidectomy can always represent an option for modern surgery of thyroid disease. METHODS: One hundred-thirty-seven patients underwent operations for thyroid disease during an 11-year period. Fifty-three patients underwent total thyroidectomy, 27 affected by a benign pathology and 26 by a malignant one. RESULTS: Both short term and long term results have been analysed. CONCLUSIONS: In conclusion, the authors affirm that a complete resection of the gland is mandatory for the surgeon in the treatment of malignant diseases because the primary aim for oncologic surgery of the thyroid is the reduction of local recurrence and the increase of survival. As far as benign diseases are concerned, some surgeons affirm that morbility of non-total operations is lower than total thyroidectomy; anyway, the authors affirm that the most important factor to prevent morbility after total thyroidectomy is an appropriate surgical technique. In this way, both the identification of parathyroid glands and the identification preparation of recurrent nerve are considered the most valid method to prevent lesions. Another factor in favour of total thyroidectomy is represented by the risk of carcinoma on the residual tissue. PMID- 11479434 TI - [Reproductive history, contraceptives and cigarette smoke as risk factors for cancer of the thyroid in women. Case-control study]. AB - BACKGROUND: Evaluation of the influence of hormonal and reproductive factors and the role of cigarette smoking in the onset of thyroid carcinoma. METHODS: Comparison between a group of 78 female patients all living in the district of Lazio, operated for thyroid follicular or papillary carcinoma at the Third Clinical Surgery Dept., Policlinico Umberto I, Rome (Italy) from 1990 to 1997, and a group of 150 women free from neoplastic and/or hormonal pathology, recruited by the compilation of a questionnaire. RESULTS: Cigarette smoking can be associated with risk reduction of developing thyroid neoplasia. On the contrary, no risk variation has been associated with the number of normal pregnancies, with pregnancy interruption both spontaneous and voluntary and with anthropometric characteristics of the analysed individuals. The first pregnancy at very young age and the use of contraceptives seem to determine a risk increase of thyroid cancer, at the limit of statistical significance. CONCLUSIONS: The antiestrogenic action of cigarette smoking exerts a protective action for thyroid carcinomas. Spontaneous or volontary interruption of pregnancy did not show a significant effect as risk factor. PMID- 11479435 TI - Parathyroid carcinoma. A concise review. AB - Parathyroid carcinoma is a very rare malignancy and the least common among endocrine malignat tumors. Its etiology is still unclear and some hypotheses have been suggested: previous external radiation to the neck, pregression from benign to malignant lesion, familial hyperparathyroidism. An early accurate diagnosis is very important since surgery is the most effective therapeutic approach to parathyroid carcinoma. Prognosis is quite variable and stress is laid on the early diagnosis and radical surgery which seem to be most favorable prognostic factors. An accurate follow-up should include frequent measurements of serum calcium and PTH levels to promptly detect recurrence or metastases which are usually associated with severe hyeprcalcemia. PMID- 11479436 TI - [Insulin therapy]. AB - Large randomised, controlled clinical trials have clearly shown that, both in type 1 (DCCT) and in type 2 (UKPDS, Kumamoto Study), micro and macrovascular complications are largely preventable with intensive insulin or oral hypoglycemic therapy and optimal metabolic control. Intensive insulin therapy can be performed in selected cases through continuous subcutaneous insulin infusion with a pump. However, almost equally good results can be obtained with a regimen of multiple injections based on a rational association of fast acting and slow acting insulin preparations. The present availability of fast acting insulin analogues (lispro and aspart insulin), characterised by aminoacid substitutions in the C-terminal portion of B-chain and a fast absorption rate from the subcutaneous tissue, is of great advantage in this context. Fast-acting analogues restore prandial insulin peaks while multiple doses of NPH restore basal insulin levels for the control of fasting blood glucose levels. New protracted acting insulin analogues and premixed preparations of fast acting analogues and NPH will soon be available. Pharmcokinetics of the protracted - acting analogue glargine seems particularly satisfactory. Glargine is present in vials in acid solution and has a neutral isoelectric point. Therefore, when injected, glargine precipitated in the subcutaneous tissue and is reabsorbed very slowly in the following 20-24 hours. Moreover, new fatty acid derivatives of insulin analogues are now under experimentation. The protraction of action of this kind of insulin is due to its binding with serum and interstitial albumin and subsequent slow release. Multiple insulin regimens are available for different types of diabetes, different degrees of insulin deficiency and peculiar clinical features of individual patients. However, in any case, insulin therapy should aim to near - normalise not only mean blood glucose and glycated hemoglobin values but also postprandial blood glucose level which represent an independent risk factor an for diabetic complications. PMID- 11479437 TI - [Coronary artery disease. Update and prospects of radiologic imaging with CT and MR]. AB - The new diagnostic options offered by the continuous development of CT and MR imaging of the heart must be periodically reviewed. This review article aims to outline the main developments in CT and MR technology and to evaluate the results and indications in CAD (coronary artery disease) patients. PMID- 11479438 TI - [Papillary lesions of the breast: diagnostic imaging and contribution of percutaneous needle biopsy with 14G needle]. AB - PURPOSE: To evaluate the presence of suggestive mammographic, US, color-Doppler, RM findings of 33 PLB and to show the accuracy of the large-core biopsy in evaluating these lesions. MATERIAL AND METHODS: A retrospective review of imaging guided large-core biopsy of 860 consecutive lesions revealed that PLB were diagnosed in 31/33 cases and 3 were suspicious papillary lesions. Surgical correlation was available for all these lesions. Mammography and US were performed in all patients, Color-Doppler in 13/33 and MR in 10/33. RESULTS: Histological findings by percutaneous biopsy demonstrated 26 (79%) benign, 3 (9%) atypical, and 4 (12%) malignant lesions. Histological findings after surgery confirmed the diagnosis for benign and malignant lesions, while of the 3 atypical lesions, 1 was benign and 2 were malignant. One encysted papillary carcinoma in situ at core-biopsy was classified as invasive papillary carcinoma after surgery. PLB were usually found (52%) in subareolar location and the mean size was 17 mm (range 5-60 mm). The most frequent mammographic appearance of benign PLB was of a well-defined (71%), oval (53%) mass. The microcalcifications had variable features; they were isolated in 3/27 (15%) cases and associated with masses in 4/27 (20%). The mammographic finding of papillary carcinoma was of a well-defined (50%) or ill-defined, oval (50%) or lobulated (50%) mass, but never of a spiculated mass. US finding of the benign PLB most commonly showed a well-defined (84%), oval (84%), complex solid/cystic (52%) mass with frequently (60%) posterior enhancement. US finding of papillary carcinoma was of a well-defined (50%) or ill-defined (50%), oval (50%) or lobulated (50%) mass, most commonly solid-inhomogenous-hypoechoic. Color-Doppler showed high blood flow in 8/10 benign PLB and in 2/3 malignant PLB. Contrast-enhanced MR imaging demonstrated usually well-circumscribed, round masses (71%). The intensity/time curve showed marked focal enhancement (peak signal intensity over 70% at the first minute) in both benign and malignant lesions. DISCUSSION: Often the patients with PBL were symptomatic (for presence of nipple discharge or palpable mass). 31/33 papillary lesions identified at the subsequent imaging-guided large-core biopsy and in the 3/33 remaining lesions percutaneous core-biopsy required a subsequent surgical biopsy for the atypical papillary lesions. US proved to have the highest sensitivity, showing the suggestive feature of a frond-like mass within a dilated duct, and color-Doppler demonstrated high blood flow (which should be considered in differential diagnosis of galactocele). Mammographic finding of papillary lesions was often consistent with benign lesions (fibroadenoma, cyst). MR confirmed the high vascularization of these lesions, showing marked enhancement of the solid component. CONCLUSIONS: US, with Color-Doppler, proved to be the most useful examination for the identification and demonstration of the solid component of these lesions, which, observed further diagnostic investigation. As no definite mammographic, sonographic or RM pattern could be identified to differentiate between benign and malignant PLB, core-biopsy was required. Percutaneous biopsy has shown to be reliable in the diagnosis of benign and malignant PLB (without any false negative): infact, any atypical lesions require surgical examination. The framing of benign and malignant PLB with imaging and core-biopsy was useful because the frequent association of benign PLB with concurrent or subsequent breast carcinoma suggests surgical excision and radiological follow-up. PMID- 11479439 TI - [Geometrical analysis of benign and malignant breast lesions]. AB - PURPOSE: The aim of this study was to describe a computer-assisted method based on fractal dimension and the coefficient of roundness, two mathematical descriptors of irregularly shaped objects, in order to discriminate benign and malignant mammographic lesions. MATERIAL AND METHODS: Twenty-three digitized mammograms were classified as containing benign lesions (n=12) or as containing malignant lesions (n=11) on the basis of readings by two independent radiologists. Morphometrical and fractal analysis of the breast lesions were automatically performed by a computer-assisted image analysis system. RESULTS: The method shows, for the first time, two parameters that allow the classification of irregularly shaped objects, such as benign and malignant breast lesions. A significant increase was obtained when comparing the fractal dimension and the coefficient of roundness of benign versus malignant mammographic lesions. DISCUSSION AND CONCLUSIONS: Benign and malignant breast lesions are characterized by complex morphologies. Complexity is the main property of all biological systems, including natural and pathological structures. In this study, we showed that fractal geometry allows quantitative measurement of the complex morphology of benign and malignant mammographic lesions. Furthermore, this mathematical approach may be helpful for the study of the non-linear dynamic processes involved in breast carcinogenesis. PMID- 11479440 TI - [Usefulness of chest X-ray during cystic fibrosis exacerbation in adult patients]. AB - PURPOSE: Verify the real usefulness of chest X-ray during clinical exacerbation of cystic fibrosis (FC). MATERIAL AND METHODS: The study was based on a group of 46 adult patients affected by FC. For every subject we selected one or more pair of chest films, of which the first was used as reference image (To), whereas the second (T1) was selected among the following controls, either in course of clinical obvious exacerbation, or during another routine control. The 64 pairs of chest films (32 representing phases of clinical stability, 32 documenting evolution towards clinical deterioration) were subjected to evaluation by two radiologists, who were first asked to judge if stability, improvement or worsening of the overall radiographic picture could be observed; then to estimate the improvement, worsening, stability or absence of the 12 radiographic signs, selected among those more frequently correlating with the acute phase of disease. RESULTS: The comprehensive radiological evaluation, which shows a statistically significant difference (p=0.012) between the two groups considered, weakly correlates with the patient clinical status. Among the patients with exacerbation, the radiographic picture appeared worsened in only 18/32 cases (sensibility 56%, specificity 78%), while on the contrary 7/32 clinically stable patients exhibited a radiographic deterioration. Among the considered radiographic-sings, some were exclusively observed during exacerbation (specificity 100%), but with low sensitivity (pneumothorax: 6%, increment of bronchiectasis: 9%, air space disease 22%); the most common alterations (mucoid impactions and bronchial wall thickening) were observed in both groups of patients. DISCUSSION AND CONCLUSIONS: Our results demonstrate that there is not a precise correlation between the radiographic picture and the clinical manifestations of exacerbation and confirm the poor usefulness of chest X-ray in such a phase of disease. Chest X-ray is needed to exclude pnemothorax or extensive air space disease, rather then to accurately diagnose exacerbation. PMID- 11479441 TI - [Staging and re-staging of soft tissue sarcoma using MRI. Usefulness of contrast media]. AB - PURPOSE: This study aims to evaluate the usefulness of paramagnetic contrast medium (Gadolinium) in the staging and re-staging of soft tissue tumors using Magnetic Resonance Imaging (MRI). MATERIAL AND METHODS: Sixty patients affected by soft tissue sarcoma of different histotype were retrospectively evaluated. Age ranged from 23 to 78 years. Data were obtained from the musculoskeletal tumor database of our Department of Radiology and cases from the last 7 years (1993 1999) entered this study. All the patients were submitted to ultrasound (US) and to routine and enhanced MR examinations (SE T1-weighted and Fast-SE T1 and T2 weighted before and after fat saturation pulse; GE T2-weighted sequences). At least two different scan planes were obtained depending on the major axis of the lesion. Once imaging information had been obtained all the patients underwent surgery and histology. After surgery, the mean follow-up period was 18 months. Paramagnetic contrast medium (0,2mmol/Kg) was administered in all the MR follow up examinations. RESULTS: Lesions included adipose histology in 15 cases; muscular histology in 12 cases; fibro-hystiocitic histology in 20 cases; synovial histology in 8 cases; mixed histology in 5 cases. Excluding lesions with mainly adipose content showing obvious high signal intensity on T1-weighted sequences, the other lesions rarely presented MR features specific to histological category also after contrast medium administration. In all the cases MRI allowed an accurate definition of the lesion boundaries to exclude or demonstrate neighbor region invasion. In 11 cases, MRI was able to demonstrate the presence of a recurrent tumor. In 2 out of 11 cases, recurrences were detected out only after intravenous administration of Gadolinium, plain MRI having failed to recognize them. DISCUSSION AND CONCLUSIONS: Despite the absence of specific MRI signs of the different soft tissue tumor histotypes also after Gadolinium administration, MRI remains the best imaging technique to establish the exact morphology of the lesions and to establish the invasion of the neighbor regions by the tumors. The presence of regional spreading of the tumor has to be considered a sign of malignancy. During the follow-up, the use of Gadolinium allows the MR examinations to reach high sensitivity levels and makes it also possible to recognize local recurrences of a very small size. Considering our experience we recommend, whenever a follow-up of resected soft tissue tumors is requested, to perform MRI examination before and after intravenous Gadolinium administration. PMID- 11479442 TI - [Bifid median nerve in the carpal tunnel: integrated imaging]. AB - PURPOSE: Carpal Tunnel Syndrome (CTS) is a neuropathy caused by compression of the median nerve at the level of the wrist, which is caused by different pathologic conditions. In some cases it has been associated with anatomic variations of the median nerve, mainly with the early duplication of the nerve inside the carpal tunnel (III Group of Lanz). The treatment of CTS is mainly surgical consisting in decompression by transection of the transverse carpal ligament, which is better performed by endoscopic release. The aim of our study was to evaluate the possibility to detect this anatomic variant by integrated imaging and assess the value of this information for planning treatment. MATERIAL AND METHODS: We report 6 cases of bifid median nerve examined by Ultrasonography (US) using mechanic sectorial and linear array transducers operating at 10 to 13 MHz, and by MR imaging using 0.2 T equipment. RESULTS: In all cases US transverse scans showed two adjacent oval formations with a structure similar to that of the median nerve. CONCLUSIONS. MR imaging confirmed the sonographic findings in all cases. All patients underwent open surgical treatment by decompression of the median nerve; in all cases an early duplicated median nerve was confirmed. PMID- 11479443 TI - [Spiral-CT with multiplanar reconstructions (MPRS) in the evaluation of ureteral neoplasms: preliminary results]. AB - PURPOSE: Urothelial tumors of the upper urinary tract (renal pelvis and ureters) are rare: the most common of these are Transitional Cell Carcinomas (TCC), usually localized in the bladder (75% of cases), whereas only 7-10% arise in the renal pelvis and 2,5-5% in the ureter. Multicentricity is not infrequent and synchronous or metachronous tumors have been reported to develop in 11-13% of patients with upper urinary tract cancer and 2-4% of those with bladder cancer. The presence of additional lesions can change patient management. Clinically these tumors may present with hematuria or flank pain, but sometimes there are no specific clinical findings. Radiological studies play an important role in tumor detection and staging and in follow up. The diagnosis is usually performed by IVU and US, while CT is generally employed in the staging. Our purpose was to verify the role of Spiral CT with Multiplanar Reconstructions (MPRs) in the evaluation of ureteral tumors. MATERIAL AND METHODS: Nine patients with ureteral neoplasms were examined with a single detector spiral-CT, using the following parameters: 120 kV, 200 mA; 5 mm collimation, pitch 1.4, slice reconstruction every 5 mm; image acquisition 2-5 minutes after injection of 120 cc of contrast medium (300 mgI/ml). All the exams were transferred to a separate workstation to obtain curvilinear MPRs. RESULTS: Spiral CT on the axial plane visualized 4 urothelial tumors as concentric ureteral wall thickening; 3 as intraluminal soft tissue masses, with involvement of almost the entire urinary tract, as far as the bladder in 1 case. In other 2 patients there were synchronous lesions which presented both patterns (wall thickening and intraluminal mass). In all cases the lesions exhibited moderate enhancement after c.m. injection. Medium- to high grade hydronephrosis above the tumors was always present. On MPRs both ureteral involvement and dilated, urine-filled upper urinary tract were clearly represented on coronal and sagittal oblique planes; these images allowed to visualize the craniocaudal extension of short or long lesions, and to detect multicentricity. DISCUSSION: The radiological diagnosis of ureteral tumors is usually performed by IVU, US and CT. IVU allows to visualize the tumors as filling defects or irregular narrowing of the ureteral lumen. IVU however is unable to show the real extent of the tumor in obstructive disease or in nonfunctioning kidney. US visualizes the lesion as an echoic mass in the lumen, although it is sometimes difficult to differentiate neoplastic lesions from other filling defects and to visualize the entire ureteral tract. CT is generally employed in the staging of ureteral tumors. CT findings are usually very useful for distinguishing ureteral neoplasms from other filling defects based on their attenuation and enhancement characteristics. Spiral CT with MPRs allows to obtain panoramic views of the urinary tract; these images are more effective than those of IVU, because of their capability to show the longitudinal extension of the lesion and the presence of multicentric tumors. Furthermore with MPRs it is possible to visualize the ureteral tract distal to the lesion, overcoming the limitations of IVU in nonfunctioning kidney or obstructive disease. CONCLUSIONS: Spiral CT with MPRs is useful in the evaluation of ureteral tumors as it is capable of visualizing the whole extension of the lesion and the presence or absence of multicentric lesions, which can change patient management. PMID- 11479444 TI - [Helical CT without contrast media as alternative to other methods in urolithiasis]. AB - PURPOSE: During the last five years the use of unenhanced helical CT (HTC) has been proposed as a preliminary diagnostic approach in patients with typical renal colics or with nonspecific flank pain. The aim of this study was to describe our experience in this field. MATERIAL AND METHODS: 130 consecutive patients (82 men and 48 women, with an average age of 54 years) were studied; 94 of them presented typical renal colics, while 36 presented acute flank pain. The parameters used were: 5-mm-thick sections, a pitch of 1.6, a reconstruction interscan spacing at 3 mm, an examination area extending from the kidneys to the base of the bladder. Depending on the case, examinations were carried out using urography in the instrumental or surgical pretreatment phase, ultrasonography to check urinary dilatation and juxtavesical calculi, pyeloMR, ureteroscopic extraction, surgical operation, extracorporeal lithotripsy, and finally the expulsion of the calculi was checked. RESULTS: 81 cases of urolithiasis, 6 cases of renal abnormalities and other renal pathologies, 23 cases of extraurinary pathologies correlated to the symptoms, 5 cases of extraurinary pathologies not correlated to the symptoms and 15 normal cases, were diagnosed. The results obtained were a) a sensitivity of 96.8%, a specificity of 98.4% and a diagnostic accuracy of 97.6% in identifying ureteral calculi; b) a diagnostic accuracy of 100% in identifying urinary tract dilatation; c) a sensitivity of 100% and a specificity of 96.7% in determining the level of obstruction; d) a sensitivity of 92.9% and a specificity of 100% in determining the cause of obstruction. CONCLUSIONS: Confirming the data in the medical literature, helical CT has yielded for more reliable results than the other procedures in identifying the following: calculi, acute obstruction of the urinary tract and other urinary and extraurinary pathologies correlated to the symptoms. Therefore helical CT, where available, must be accepted as the method of preliminary evaluation in all patients with typical renal colics or with nonspecific flank pain. PMID- 11479445 TI - [Magnetic resonance imaging in the evaluation of parametrial invasion of carcinoma of the cervix uteri: optimization of the study protocol]. AB - PURPOSE: To determine the efficacy of three different MR sequences in the evaluation of parametrial invasion by early-stage cervical cancer. MATERIAL AND METHODS: Eighteen consecutive patients with cervical cancer clinically assessed as stage IB1 underwent MR imaging examination with the use of the following sequences: FSE T2-weighted, FSE fat-suppressed T2w, and SE fat-suppressed Gadolinium-enhanced T1w. In all cases, the presence or absence of parametrial invasion on both sides per each sequence used was evaluated. Subsequently all the sequences have been considered together for the evaluation of tumor invasion. Gold standard of the study was the histopathologic analysis of the surgical specimens. RESULTS: At histological examination, parametrial invasion by tumor was found in 6 out of 36 parametria evaluated. The accuracy achieved with each of the sequences used was as follows: 94% with FSE T2w; 86% with FSE fat-suppressed T2w; and 67% with SE fat-suppressed Gadolinium-enhanced T1w. The simultaneous evaluation of all 3 sequences obtained an accuracy level similar to that achieved with FSE T2w. The difference between the accuracy of T2w sequences and that of fat-suppressed contrast-enhanced T1w sequences was statistically significant (p<0.01). DISCUSSION AND CONCLUSIONS: Our data suggest that the MR imaging protocol for the evaluation of parametrial tumor invasion could be restricted to FSE T2w sequences. These proved to have the highest negative predictive value (97%) which allows a reliable selection of patients who can be surgically treated. PMID- 11479446 TI - Iohexol does not influence the levels of blood serum cation electrolytes during intravenous pyelography. AB - PURPOSE: The influence of the nonionic contrast agent iohexol on blood pH and the cation electrolytes Ca, Na, K and Mg was studied in vivo in humans during the intravenous pyelography. MATERIAL AND METHODS: A population of 28 patients without renal function impairment or electrolyte imbalance was divided into two equal groups. The first group received an intravenous injection of 0.6 mL Omnipaque per kg of body weight (0.388g iohexol/kg and 0.18 g iodine/kg). The second group received a double dose of Omnipaque (1.2 mL iohexol/kg; 0.776g iohexol/kg and 0.36g iodine/kg). Blood samples (5 mL) were taken before the injection of the contrast medium and 5, 10 and 20 minutes (marking the end of the examination) after the injection. Blood pH was also determined immediately after each sample collection, while electrolytes were measured in the laboratory. Systolic and diastolic blood pressure were also determined simultaneously with each sample collection using a mercury manometer. RESULTS: Iohexol in simple and double doses did not have any statistically significant effect either on blood pH or on blood serum electrolytes during intravenous pyelography. Blood pressure, both systolic and diastolic, remained stable in all the patients during the examination. CONCLUSIONS: Our results show that the nonionic contrast medium iohexol at the doses used in routine intravenous pyelography does not have any influence on blood pH and serum cardioactive cation electrolytes in humans. An electrolyte imbalance is always a risk factor that may contribute to the development of cardiovascular side effects. This factor is eliminated when iohexol is administered intravenously instead of an ionic agent. PMID- 11479447 TI - [Non-surgical treatment of aortic aneurysms and dissections: indications, strategies and outcome]. AB - PURPOSE: The endovascular treatment of the thoracic aorta is an effective alternative to open surgical repair and offers a therapeutic option even to patients at high risk for surgery. Our experience in the treatment of different diseases of descending thoracic aorta is reported. MATERIAL AND METHODS: Between July 1997 and January 2001, 50 patients were selected for endovascular treatment: 36/50 patients presented high risk for conventional surgery. Six patients presented clinical and imaging features suggesting impending rupture and were treated on emergency basis. The stent-graft prosthesis was individually manufactured or selected on the basis of spiral CT or MRI measurements. RESULTS: Endovascular stent positioning and deployment was technically successful in 49 cases. In one patient the tortuosity of the aortic arch prevented graft deployment. Complete aneurysm exclusion was achieved in 48 cases as assessed by post-procedure angiography and TEE. One proximal endoleak was noted and surgical conversion was performed 40 days later. There were no intraoperative mortality or complications. One patient presented extension of dissection at the 8th postoperative day and required of surgical repair. CT scan showed an endoleak in 4 cases that sealed spontaneously in three cases while the fourth case was treated by graft extension. In the long term two secondary endoleak were observed (12 and 24 months after the procedure). CONCLUSIONS: Endovascular stent-graft repair provides a less invasive opportunity to patients affected by thoracic aortic disease. Careful cases selection is the first postulate for the efficacy and safety of the procedure. PMID- 11479448 TI - [Unresectable hilar cholangiocarcinoma: combined percutaneous and radiotherapic treatment]. AB - PURPOSE: Evaluate, in patients with inoperable hilar CLCA, the efficacy of multimodality treatment (brachiotherapy, chemotherapy, external radiotherapy and endoprosthesis positioning) in terms of survival, quality of life and cost/benefit compared to palliative surgical treatment. MATERIAL AND METHODS: 20 patients with inoperable hilar CLCA were evaluated. Ten were considered for combined palliative and radiotherapy treatment according to the following protocol: percutaneous colangiography followed by positioning of right and left internal biliar drainage (10-12 F); intraductal brachiotherapy using Ir-192 needles was performed (7 Gy). A second administration was after 7 days. Total dose was 14 Gy; the biliar drainages were then replaced by endoprosthesis (12-14 F) and left in position for 3 months in order to model the post-attinic fibrosis and to prevent stenosis; external radiotherapy was administered starting 15 days after last brachiotherapy treatment (26 administrations (180 cGy) in 5 weeks, total 46 Gy); chemotherapy (5 FU: 350 mg/mq/die) for 5 days during the first and the fifth week of external radiotherapy; biliar endoprosthesis were removed by endoscopic and, or transhepatic mode after 3 months from end of therapy, verifying. Cholangiography assessed the patency of the biliar duct. Metallic stents were placed if results were not satisfactory; follow-up was by: hepatic lab work-up, tumoral markers, US or TC evaluated disease progression. RESULTS: In 5 of the 10 patients considered for the combined treatment, 5 patients completed the protocol and 5 were treated only with brachiotherapy because of deterioration of clinical conditions. All patients had initial complete remission of jaundice although it recurred with disease progression that led to death. No acute post radiotherapy complication was observed. Digestive hemorrhage (chronic post radiotherapy complication) was the cause of death in one patient. DISCUSSION AND CONCLUSIONS: Good results were observed in the patients that completed the protocol, mean survival was 7,5 months instead of 1,75 months as in patients that underwent only percutaneous drainage. Better results may be obtained taking in consideration the poor clinical conditions of the patients at the time of diagnosis. The proposed therapeutic protocol requires an average hospitalization of 10-15 days instead of 15-20 days as for palliative biliar-digestive deviation, being less invasive and thus associated to lower morbidity and no mortality. PMID- 11479449 TI - [Association of cerebral and spinal vascular artero-venous malformations: report of a case]. PMID- 11479450 TI - [LIP (Lymphoid Interstitial Pneumonitis) and HIV infection in children. Case report]. PMID- 11479451 TI - [Intrahepatic portal vein gas. Good prognosis in patients with gastric dilatation. Case report]. PMID- 11479452 TI - [Epithelioid leiomyoblastoma of the greater omentum: a case report]. PMID- 11479453 TI - [Duodeno-caval fistula: CT findings. Case report]. PMID- 11479454 TI - Desperate diseases, desperate measures: tackling malignant hypertension in the 1950s. AB - BACKGROUND: The conquest of malignant hypertension is one of the most important medical achievements of the second half of the 20th century. As we enter the new millennium, it is critical to examine the efforts that have led to our ability to treat this once incurable disease. METHODS: Review was performed of the literature from 1900 to the 1950s regarding the etiology, clinical evaluation, and treatment of hypertension, focusing on malignant hypertension. RESULTS: Fifty years ago, in a time of sparse treatment options, the occurrence of malignant hypertension was a dreaded event that taxed the aptitude of the clinician. Confronted with an "extreme disease," physicians chose to use "extreme methods of cure" in conformity with the teaching of Hippocrates. In the 1950s malignant hypertension was treated with such drastic measures as rice diet, sympathectomy, and intravenous pyrogens. CONCLUSIONS: In the practice of medicine today, while work is being done to reassert biomolecular mechanisms, we still face patients who have reached the end stages of failure and manifest devastating morbidity. These patients are subjected to "extreme therapies" reminiscent of those that surrounded malignant hypertension in the past. In an era when adequate treatment of hypertension has become a reality for so many patients, it is appropriate to give credit to those who paved the way to such great progress. PMID- 11479455 TI - Comparative efficacy of fibrinogen and platelet supplementation on the in vitro reversibility of competitive glycoprotein IIb/IIIa (alphaIIb/beta3) receptor directed platelet inhibition. AB - BACKGROUND: Platelet surface glycoprotein (GP) IIb/IIIa (alphaIIb/beta(3)) receptor inhibition, by preventing fibrinogen binding and platelet aggregation, concomitantly attenuates arterial thrombotic capacity and impairs protective hemostasis, 2 divergent platelet-dependent processes. Because the currently available Food and Drug Administration-approved small molecule GP IIb/IIIa receptor antagonists are considered "competitive" inhibitors and because there is limited information on the reversibility of platelet inhibition by fibrinogen or platelet supplementation, the following series of in vitro experiments were performed. METHODS AND RESULTS: Washed platelets from 24 healthy volunteers were suspended in Tyrodes buffer and incubated with achievable (in vivo) steady-state concentrations of either tirofiban or eptifibatide before activation with TRAP (thrombin receptor agonist peptide) (15 micromol/L). Platelet aggregation was inhibited by 40% to 50%, but reversal was achieved by fibrinogen supplementation in a concentration-dependent manner. In a separate series of in vitro experiments, platelet inhibition exceeding 90% was established with tirofiban (average concentration 9.28 microg/L) and eptifibatide (average concentration 95.4 microg/L). Recovery of platelet aggregation to at least 50% was achieved after the addition of fibrinogen (0.76-0.80 g/L), platelets (2.4 x 10(11)/L), or their combination. There was an inverse relationship between plasma baseline fibrinogen and the amount of supplemental fibrinogen required to restore platelet aggregability (r = -0.60, P <.01). CONCLUSIONS: The reversibility of GP IIb/IIIa directed platelet inhibition is influenced by cell surface receptor availability and the intrinsic pharmacodynamic mechanism of action. Fibrinogen supplementation with fresh frozen plasma or cryoprecipitate either alone or in combination with platelet transfusion, represents an important and readily available treatment consideration for restoring hemostatic potential and managing major hemorrhagic complications associated with the administration of small molecule competitive GP IIb/IIIa receptor antagonists. PMID- 11479456 TI - The A-to-Z Trial: Methods and rationale for a single trial investigating combined use of low-molecular-weight heparin with the glycoprotein IIb/IIIa inhibitor tirofiban and defining the efficacy of early aggressive simvastatin therapy. AB - BACKGROUND: The A-to-Z Trial is an ongoing international, multicenter, randomized study designed to investigate 2 issues concerning contemporary care of patients with acute coronary syndromes (ACS). The first issue is whether the use of low molecular-weight heparin versus unfractionated heparin affects outcomes and safety when used as a therapy adjunctive to baseline treatment with tirofiban and aspirin in patients with non-ST-elevation (nSTE) ACS. The second issue is whether early use of an aggressively dosed statin is superior to a current trial-based "accepted care" regimen of a lower-dose statin started 3 to 6 months after an acute event. METHODS: The study is conceptually and functionally divided into 2 sequential parts-the "A" Aggrastat and "Z" Zocar phases. The primary A-phase end point is a composite of all-cause mortality, myocardial infarction (MI), and documented refractory ischemia at 7 days. Both nSTE-ACS patients from the A phase and patients with ST-elevation ACS who meet specific risk criteria are eligible to enter the subsequent "Z" (Zocor) chronic phase (Z phase). The primary end point of the Z phase is a composite of cardiovascular death, MI, readmission for ACS, and stroke. The trial will continue until 970 primary events have occurred in the Z-phase population. CONCLUSION: This trial is evaluating 2 temporally connected sequences of phamacotherapy for ACS. At completion, trial results will provide definitive evidence regarding efficacy and safety of early, intensive statin therapy and better define the role of low-molecular-weight heparin in patients with nSTE ACS. PMID- 11479457 TI - Conducting pediatric cardiovascular trials. Introduction. PMID- 11479458 TI - Pediatric issues and diseases. PMID- 11479459 TI - End points for cardiovascular drug trials in pediatric patients. PMID- 11479460 TI - Ethical issues in pediatric trials. PMID- 11479461 TI - Randomized comparison of a novel anticoagulant, vasoflux, and heparin as adjunctive therapy to streptokinase for acute myocardial infarction: results of the VITAL study (Vasoflux International Trial for Acute Myocardial Infarction Lysis). AB - BACKGROUND: Vasoflux is a low-molecular-weight heparin derivative that inhibits factor IXa activation of factor X and catalyzes fibrin-bound thrombin inactivation by heparin cofactor II. We studied whether vasoflux improves the results of thrombolysis with streptokinase for acute myocardial infarction. METHODS AND RESULTS: We randomized 277 patients with acute myocardial infarction to standard intravenous unfractionated heparin (UFH) or intravenous vasoflux 1, 4, 8, or 16 mg/kg as a bolus followed by 1, 4, 8, or 16 mg/kg per hour infusion, on top of streptokinase and aspirin, until angiography at 90 minutes. Patency and corrected Thrombolysis in Myocardial Infarction (TIMI) frame count were studied at 60 and 90 minutes. Rates of TIMI grade 3 flow with vasoflux at any dose (35% to 42%) were not different from UFH (41%) at either time point, nor was the corrected TIMI frame count. However, there was an excess of bleeding in the patients randomized to vasoflux 8 or 16 mg/kg: 78% and 71%, compared with 53% for UFH (P =.004 and.043, respectively). Major bleeding was observed in 13% and 28% at these vasoflux doses compared with 8% with UFH (P =.558 and.01, respectively). CONCLUSION: At doses that increase the risk of bleeding, the addition of vasoflux to streptokinase and aspirin did not lead to improved patency rates compared with UFH. Targeting factor IXa and heparin cofactor II may not be a useful adjunct to thrombolysis. PMID- 11479462 TI - A randomized trial confirming the efficacy of reduced dose recombinant tissue plasminogen activator in a Chinese myocardial infarction population and demonstrating superiority to usual dose urokinase: the TUCC trial. AB - BACKGROUND: Reports from Japan suggest effective myocardial infarction (MI) treatment in Asian patients with much lower doses of tissue plasminogen activators (tPA) than used in European and American regimens. Because increasing doses of fibrinolytics lead to increased bleeding complications, identification of patients who respond to reduced doses is of importance. We conducted a trial in the People's Republic of China in which reduced-dose recombinant tPA was compared with the standard local therapy, urokinase. METHODS: Four hundred patients with acute MI within 12 hours of symptom onset were to be randomized to an 8-mg bolus of recombinant tPA followed by a 42-mg 90-minute infusion or 1.5 million units of urokinase as a 30-minute infusion. Patients received aspirin and heparin and underwent angiography to determine infarct artery patency 90 minutes after the start of therapy. RESULTS: The Data and Safety Monitoring Board recommended premature termination after 342 patients were recruited. Infarct artery patency (grade 2 or 3) occurred in 79% of patients receiving recombinant tPA and in 53% of patients receiving urokinase (P <.001); Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow was 48% and 28%, respectively (P <.001). The higher-patency-rate recombinant tPA growth had better posttreatment left ventricular ejection fractions, 58.6% versus 54.7%, P <.01. Adverse events were infrequent and not significantly different in the 2 groups. CONCLUSIONS: This study confirms that a substantially lower dose of recombinant tPA is effective in Asian patients compared with that required in Western patients even after consideration of body weight. Specific dose-response studies should be performed with fibrinolytic regimens to avoid overdosage with its attendant risks of excess bleeding. PMID- 11479463 TI - Role of angiotensin II type 1 receptor in the regulation of cellular adhesion molecules in atherosclerosis. AB - BACKGROUND: Inflammation is a central feature of coronary artery disease (CAD) that is characterized by increased expression of cellular adhesion molecules with the exception of L-selectin. L-selectin is a leukocyte adhesion molecule that is rapidly shed after leukocyte activation so that it appears to be decreased in CAD. The renin-angiotensin system (RAS) is implicated in atherogenesis and up regulates these molecules. OBJECTIVES: The aim of this study was to investigate the effect of angiotensin type 1 (AT1) receptor antagonism on serum and leukocyte adhesion molecule expression in patients with CAD. Blood samples were collected from 31 patients before and after 8 weeks of treatment with losartan (44 +/- 2 mg/d, mean +/- SE), an AT1 receptor antagonist. We measured serum intercellular adhesion molecule-1, vascular cell adhesion molecule-1, endothelial-leukocyte adhesion molecule, and C-reactive protein (CRP). By flow cytometry, we also measured the expression of leukocyte CD11a, CD11b, CD11c, CD18, CD31, CD49d, and CD62L (L-selectin) in 13 patients. RESULTS: Treatment with losartan decreased systolic blood pressure (141 +/- 3 vs 135 +/- 4 mm Hg, P =.04) and increased plasma renin activity (1.2 +/- 0.4 vs 2.7 +/- 0.5 ng/mL/h, P =.001). There was a significant increase in L-selectin expression on monocytes (86 +/- 6 vs 118 +/- 10 MESF units, P =.007), lymphocytes (52 +/- 10 vs 79 +/- 8, P =.01), and granulocytes (124 +/- 7 vs 156 +/- 18, P =.056). However, there were no changes in the other leukocyte and serum adhesion molecules or CRP. CONCLUSIONS: These findings suggest that AT1 receptor antagonism selectively modulates L-selectin expression on leukocytes and that endogenous stimulation of AT1 receptors by the RAS contributes to the activation of leukocytes and decreased expression of L selectin in CAD. PMID- 11479464 TI - Validation of three myocardial jeopardy scores in a population-based cardiac catheterization cohort. AB - BACKGROUND: The Jeopardy Score from Duke University and the Myocardial Jeopardy Index from the Bypass Angioplasty Revascularization Investigation (BARI) have been validated but never applied to a large unselected cohort. We assessed the prognostic value of these existing jeopardy scores, along with that of a new Lesion Score developed for the Alberta Provincial Project for Outcome Assessment in Coronary Heart Disease (APPROACH), a clinical data collection initiative capturing all patients undergoing cardiac catheterization in the province of Alberta. METHODS: The predictive value of these three scores were compared in a cohort of >20,000 patients (9922 treated medically, 6334 treated with percutaneous intervention, and 3811 treated with bypass surgery). Scores were considered individually in logistic regression models for their ability to predict outcome and then added to models containing sociodemographic data, comorbidities, ejection fraction, indication for procedure, and descriptors of coronary anatomy. RESULTS: All scores were found to be predictive of 1-year mortality, especially when patients are treated medically or with percutaneous intervention. In these patients, the APPROACH Lesion Score performed slightly better than the other jeopardy scores. The Duke Jeopardy Score was most predictive in those patients undergoing coronary bypass surgery. CONCLUSIONS: Myocardial jeopardy scores provide independent prognostic information for patients with ischemic heart disease, especially if those patients are treated medically or with percutaneous intervention. These scores represent potentially valuable tools in cardiovascular outcome studies. The APPROACH Lesion Score may perform slightly better than previously developed jeopardy scores. PMID- 11479465 TI - Quality of care among elderly patients hospitalized with unstable angina. AB - BACKGROUND: Guidelines for the management of unstable angina have been published by the United States Agency for Health Care Policy and Research (currently known as the Agency for Healthcare Research and Quality); however, little information is available about the quality of unstable angina care, particularly among elderly patients. METHODS: We examined 1196 elderly Medicare-insured patients hospitalized with unstable angina (ruled out for acute myocardial infarction) at Connecticut hospitals between August and November 1995 to evaluate quality of care provided during hospitalization. Patients without therapeutic contraindications were evaluated for the use of 5 Agency for Health Care Policy and Research guideline-recommended measures: electrocardiographic examination within 20 minutes of admission, use of aspirin on admission, intravenous heparin on admission, achievement of therapeutic anticoagulation among patients provided heparin, and prescription of aspirin on discharge. RESULTS: Less than half (49.6%) of patients underwent electrocardiographic examination within 20 minutes of admission. After excluding patients with contraindications, aspirin was provided to 80.1% of patients and intravenous heparin to 59.2% of indicated patients, of whom only 43.3% achieved therapeutic anticoagulation. Aspirin was prescribed to 82.3% of eligible patients at discharge. Performance on the 5 quality measures varied widely among hospitals. CONCLUSIONS: Agency for Health Care Policy and Research guideline-recommended risk stratification and therapeutic interventions are underused in elderly patients hospitalized with unstable angina, with quality of care varying widely among hospitals. PMID- 11479466 TI - Predictors of quality of life 6 months and 1 year after acute myocardial infarction. AB - BACKGROUND: Quality of life (QOL) is an increasingly important outcome measure after hospital admission for acute myocardial infarction (AMI). However, the ability to adjust these outcomes for differences between compared groups of patients is limited because the predictors of QOL after AMI are unknown. METHODS: To identify any clinical, demographic, and psychosocial characteristics of patients at admission that were independent predictors of QOL 6 months and 1 year after AMI, we measured physical and mental QOL (Short Form-36 Physical and Mental Component summary scores) and overall QOL (EuroQol health perception scale) in a prospective cohort of 587 patients admitted at 10 hospitals in Quebec. A set of plausible multivariate linear regression models was created for each outcome measure with use of the Bayesian Information Criterion. RESULTS: Mean physical, mental, and overall QOL scores corresponding to the time immediately before admission (baseline) were 45 (95% confidence interval [CI] 44-46), 47 (95% CI 46 48), and 70 (95% CI 68-72), respectively. By 1 year, mean physical, mental, and overall QOL scores were close to baseline (45 [95% CI 44-46], 48 [95% CI 47-49], and 73 [95% CI 71-74], respectively). The predictors of physical, mental, and overall QOL were similar at 6 months and 1 year. Important predictors of physical QOL were the corresponding score at baseline, age, and previous bypass surgery (beta coefficients at 1 year: 5 [per 10-point difference in baseline score], -1 [per 10-year age difference], 5.3; 95% CIs 4 to 5, -2 to -1, -9.2 to -1.3, respectively). Predictors of mental QOL were the corresponding score at baseline and depression (beta coefficients at 1 year: 3 [per 10-point difference in baseline score], -3 [per 10-point difference in depression score]; 95% CIs 2 to 4, -5 to -2, respectively). Predictors of overall QOL included the corresponding score at baseline and age (beta at 1 year: 2 [per 10-point score difference], -3 [per 10-year age difference]; 95% CIs 1 to 3, -4 to -1, respectively). Depression was also a predictor of impaired physical and overall QOL at 6 months (beta at 6 months: -1.6 [per 10-point score difference], -5.4 [per 10-point score difference]; 95% CIs -2.9 to -0.4, -7.7 to -3.2, respectively). No variables related to treatments received in-hospital were found in the most clinically relevant models. CONCLUSIONS: These results suggest that age and psychosocial characteristics at baseline are the most important predictors of QOL after AMI. Other clinical characteristics and treatments received in-hospital do not appear to strongly affect patients' long-term perceptions of QOL. PMID- 11479467 TI - Cardiac conduction abnormalities in endocarditis defined by the Duke criteria. AB - BACKGROUND: Cardiac conduction abnormalities occur in endocarditis and have been associated with infection extension and increased mortality. There have been no prospective studies of electrocardiographic (ECG) conduction changes in endocarditis. We examined the incidence of ECG changes in a large prospective cohort with suspected endocarditis and correlated changes with echocardiographic evidence of invasive infection and mortality. METHODS: One hundred thirty-seven of 1396 (10%) suspected cases of endocarditis were classified as "definite" or "possible" by the Duke criteria and had an interpretable ECG. ECG conduction changes were classified as old (pre-existing hospitalization), new (evident on admission or developed during hospitalization), or indeterminate. New or indeterminate abnormalities were considered "ECG conduction changes." Echocardiogram results were reviewed to identify infected valves and invasive infection. RESULTS: ECG conduction changes were present in 36 of 137 (26%) patients. Patients with ECG conduction changes were more often male (69% vs 46%, P =.005) and had prosthetic valves (47% vs 23%, P <.001). There were no significant differences in microbiology results or treatment with cardiac surgery. In 76 (55%) patients, at least one infected valve was identified by echocardiography; 15 of 76 (20%) patients were determined to have evidence of invasive infection. Eight of 15 (53%) invasive infections exhibited ECG conduction changes compared with 16 of 61 (26%) isolated valve infections (P =.046). Eleven of 36 (31%) patients with ECG conduction changes died during hospitalization compared with 15 of 101 (15%) patients without changes (P =.039). CONCLUSIONS: ECG conduction changes commonly occur in endocarditis despite more sensitive diagnostic criteria and are associated with increased mortality and invasive infection. PMID- 11479468 TI - Identification of sinus node dysfunction by use of P-wave signal-averaged electrocardiograms in paroxysmal atrial fibrillation: a prospective study. AB - BACKGROUND: In patients with paroxysmal atrial fibrillation (Paf), the identification of the coexistence of sinus node dysfunction (SND) has therapeutic implications. This study sought to prospectively determine whether SND in patients with Paf would be identified by use of atrial early potential (EP), low amplitude potentials early in signal-averaged P wave. METHODS: The study population consisted of 149 patients with Paf. Signal-averaged electrocardiography was recorded with the P-wave-triggering technique. The root mean square voltage for the initial 30 MS and the duration of initial low amplitude signals < 4 microV of signal-averaged P wave were measured in the vector magnitude. The criteria of EP were defined as "the root mean square voltage for the initial 30 MS < 3.0 microV and the duration of initial low amplitude signals < 4 microV >22 MS." SND was diagnosed by use of the conventional 12-lead electrocardiography, 24-hour Holter monitoring, and bedside electrocardiographic monitoring. RESULTS: Thirty-eight of 149 patients with Paf had EP. Eighteen (47%) of 38 patients with Paf and EP had SND, whereas SND was found in only 5 (5%) of the other 111 patients with Paf without EP (P <.0001). EP gave a sensitivity of 78% and a specificity of 84% for the detection of SND in patients with Paf. CONCLUSION: EP would be useful for the identification of SND in patients with Paf. PMID- 11479469 TI - Safety and feasibility of a novel rate-smoothed ventricular pacing algorithm for atrial fibrillation. AB - OBJECTIVES: This study was conducted to establish the safety and performance of a new rate-smoothing pacing algorithm for patients with atrial fibrillation (AF). BACKGROUND: Irregularity of the ventricular response is a hallmark of AF. This irregularity may contribute to symptoms and hemodynamic compromise in patients with AF. Interventions designed to reduce irregularity have not previously been evaluated in a long-term, clinical setting. METHODS: We designed a prospective, double-blind study with randomized crossover. Patients with either paroxysmal or chronic AF whose conditions were medically refractory and who were referred for an atrioventricular node ablation procedure all underwent pacemaker implantation. Subjects were then randomly assigned to either DDD mode with the rate-smoothing algorithm (RSA) on, or to OOO mode. After 2 months they were crossed over to the other arm. RESULTS: Fourteen patients (9 with paroxysmal AF and 5 with chronic AF) were enrolled. There were no significant differences between the group randomly assigned to RSA first versus the group assigned to OOO first. The mean left ventricular ejection fraction with the RSA was not significantly different than it was in OOO mode (45.1 +/- 18.6 vs 51.9 +/- 12.3; P =.11), although some individuals with uncontrolled ventricular rates did have a large decrease in ejection fraction with rate smoothing. One developed overt heart failure. One quality-of-life instrument detected a significant improvement in the "physical limitations" domain with the rate-smoothing mode. Eleven of 14 patients preferred the RSA ON arm, and 6 of those 11 elected to defer the ablation procedure. CONCLUSIONS: Long-term rate-smoothed pacing is feasible. Because of concerns about pacing-induced heart failure in some patients with rapid ventricular rates, rate-smoothed pacing should be reserved for those who remain symptomatic despite adequate control of the ventricular rate. The RSA may help to reduce symptoms in patients with medically refractory AF; more study is required to define its efficacy in reducing symptoms and morbidity in this population. PMID- 11479470 TI - Predictors of improvement in left ventricular function after percutaneous revascularization of occluded coronary arteries: a report from the Total Occlusion Study of Canada (TOSCA). AB - BACKGROUND: The Total Occlusion Study of Canada (TOSCA) is a multicenter, randomized trial evaluating the effect of stenting with > =1 heparin-coated stent on long-term patency after percutaneous coronary intervention by balloon angioplasty of occluded coronary arteries. The purpose of the current study was to compare the effect of stenting and balloon angioplasty on global left ventricular ejection fraction (LVEF) and regional wall motion and to examine what clinical and angiographic factors may have an effect on left ventricular function in this setting. METHODS AND RESULTS: Analysis at the core angiographic laboratory of paired baseline and follow-up left ventricular angiograms, as well as target vessel patency, was possible in 244 of 410 cases. An improvement in LVEF was observed in the entire group (59.4% +/- 11% to 61.0% +/- 11%, P =.003). The LVEF change was +1.84 +/- 7.54 in the stent group (P =.009) and 1.28 +/- 8.16 in the percutaneous transluminal coronary angioplasty group (P =.085). There was no significant intergroup difference. Patients with duration of occlusion < or =6 weeks had an improvement in LVEF (+2.98 +/- 8.68, P =.0006), whereas those with an occlusion duration of > 6 weeks had no improvement (+0.48 +/- 7.01, P not significant). Multivariate analysis revealed baseline LVEF <60%, duration of occlusion < or =6 weeks, and Canadian Cardiology Society angina class I or II to be independent predictors of improvement in LVEF. CONCLUSIONS: The restoration of coronary patency of nonacute occluded coronary arteries is associated with a small but significant improvement in regional and global left ventricular function, especially in patients with recent occlusions and depressed left ventricular function. In spite of significant effect on long-term patency, stenting of nonacute coronary occlusions does not result in significantly better left ventricular function compared with balloon angioplasty in this setting. PMID- 11479471 TI - Racial differences in outcomes of veterans undergoing percutaneous coronary interventions. AB - BACKGROUND: In the era of stenting relatively little is known about racial differences in the outcomes of percutaneous interventions (PCI). The purpose of this study was to determine whether there were racial differences with respect to short- and long-term outcomes in veterans undergoing PCI. METHODS: We used the national Department of Veterans Affairs (VA) patient treatment file to identify 24,625 African American and white veterans who had PCI in VA medical centers between October 1, 1994, and September 30, 1999. Baseline demographic characteristics were obtained, as was a measure of comorbidity. Short-term outcomes included hospital mortality and same-admission coronary artery bypass surgery, and long-term outcomes were vital status and rehospitalization. Multivariate statistical methods were used to adjust for patient differences when comparing both short- and long-term outcomes for African American and white veterans. RESULTS: African Americans were 11% of veterans, and in comparison with their white counterparts had more hypertension, diabetes, and acute myocardial infarction. African Americans less often underwent stenting (44% vs 49%), although hospital mortality (2.0% vs 1.9%) and same-admission bypass surgery (1.9% vs 2.2%) rates were similar. Two-year survival was 89% in African Americans and 91% in white veterans (P =.0014), and after adjustment for covariates African Americans had slightly higher mortality rates (hazard ratio 1.11, 95% confidence interval 1.05-1.17). At 2 years almost 61% of both African American and white veterans were rehospitalized for any reason. CONCLUSION: Short- and long-term outcomes for African American and white veterans undergoing PCI in VA medical centers were similar, although African Americans underwent stenting less often. PMID- 11479472 TI - Primary renal artery stenting: characteristics and outcomes after 363 procedures. AB - BACKGROUND: Stenting improves the acute results of percutaneous balloon angioplasty for atherosclerotic renal artery stenosis. Predictors of benefit and angiographic restenosis are not well understood. We describe the technical and clinical success of renal artery stenting in a large consecutive series of patients with hypertension or renal insufficiency. We identify clinical, procedural, and anatomic factors that might influence outcome, restenosis, and survival. METHODS: Primary renal artery stenting was performed in 300 consecutive patients who underwent 363 stent procedures in 358 arteries. Angiograms were analyzed quantitatively. Clinical and angiographic follow-up data are available after a median of 16.0 months. RESULTS: At baseline, 87% of patients had hypertension, and 37% had chronic renal insufficiency. The mean age was 70 years (interquartile range 63.1-74.6) years. The stenosis was unilateral in 49% and bilateral in 48% and involved a solitary functioning kidney in 3.6%. The stenting procedure was successful in all attempts. There were no procedural deaths or emergency renal surgical procedures. Postprocedure azotemia was seen in 45 of 363 (12%) procedures but persisted in only 6 patients (2%), all of whom had baseline renal insufficiency. Systolic and diastolic blood pressures were significantly reduced (systolic blood pressure from 164.0 +/- 28.7 to 142.4 +/- 19.1 mm Hg, P <.001). At follow-up, 70% of patients had improved blood pressure control regardless of renal function. In patients with baseline renal insufficiency, 19% had improvement in serum creatinine levels at follow-up, 54% had stabilization, and 27% had deterioration. Follow-up mortality was 10% and was predicted by baseline creatinine levels (odds ratio 1.72 for each 1 mg/dL creatinine increment, 95% confidence interval 1.13-2.49) and extent of coronary artery disease (odds ratio 1.66 for each diseased coronary artery, 95% confidence interval 1.03-2.67). Angiographic restenosis was found in 21% of 102 patients overall and was less common (12%) in arteries with a reference caliber >4.5 mm (P <.01 vs caliber <4.5 mm). Neither poststenotic dilation nor severity of angiographic stenosis predicted clinical outcome. CONCLUSIONS: Primary renal artery stenting can be performed safely with nearly uniform technical success. The majority of patients with hypertension or renal insufficiency derive benefit. Follow-up mortality is 5-fold higher in patients with baseline renal insufficiency. Clinical and angiographic features did not predict blood pressure or renal functional outcome. Restenosis is more common in renal arteries with a reference caliber less than 4.5 mm. PMID- 11479473 TI - Screening cardiac ultrasonographic examination in patients with suspected cardiac disease in the emergency department. AB - BACKGROUND/OBJECTIVE: Our purpose was to evaluate the utility of a brief screening cardiac ultrasonographic (SCU) examination. We prospectively compared the SCU with conventional clinical evaluation in 124 emergency department (ED) patients with suspected cardiac disease. Furthermore, we assessed the impact and quality of SCU examinations as obtained by briefly trained ED personnel (EP). METHODS: Patients underwent clinical evaluation by an ED physician and SCU examination by a sonographer or cardiologist. Patient disposition, hospital stay length, and the number of full echo examinations were compared with the presence of significant findings on SCU. In patients who received a full echocardiogram during hospitalization, results of the initial clinical examination were compared with results of the SCU examination in the diagnosis of significant findings. A similar analysis, but with quality assessment, was performed on only those SCU examinations acquired by 4 EP. RESULTS: Of the 124 patients enrolled in the main study, 40 of 124 (32%) had significant findings on SCU. Of patients with abnormal SCUs versus normal SCUs, 16 of 40 (40%) versus 18 of 84 (21%) had hospital stay lengths >2 days (P < or =.05). Using the 36 inpatient full echo studies obtained for standard indications during hospitalization as a gold standard, initial clinical examination identified only 7 of 30 (23%) significant findings and had 16 false-positive diagnoses, whereas SCU identified 22 of 30 (73%) with 8 false positives. Although similar study results occurred with interpretation of 68 SCUs obtained by EP, quality was achieved in only 55% ED personnel versus 97% of sonographer-obtained SCUs (P <.05). CONCLUSIONS: An SCU examination detects significant findings misdiagnosed on initial clinical evaluation in the ED and provides prognostic data regarding length of hospital stay. PMID- 11479474 TI - Echocardiographic follow-up study of the Ross procedure in older versus younger patients. AB - OBJECTIVES: This study was designed to determine echocardiographic follow-up results of the Ross procedure in older adult patients with aortic valve disease. BACKGROUND: The excellent long-term results of the Ross procedure from several institutions have indicated that the pulmonary autograft may be the best available substitute for the diseased aortic valve in children and adolescents. The advantages of this operation include optimal hemodynamics and elimination of thromboembolic complications. These features may benefit older adult patients as well. METHODS: We reviewed data from 49 consecutive patients who had a Ross procedure between 1991 and 1996. Preoperative and postoperative Doppler echocardiographic studies were available for 44 patients (22 men, 22 women; mean [+/-SD] age 36 +/- 14 years) who were grouped into <40 (n = 25) and > or =40 years old (n = 19). Measurements included left ventricular diastolic volume (LVDV), mass, and ejection fraction (EF); a peak pressure gradient across autograft in the aortic position and homograft in the pulmonary position; and valvular regurgitation. RESULTS: The mean length of echocardiographic follow-up was 36 +/- 16 months. Postoperatively, there was a reduction in LVDV and left ventricular mass in both age groups: 153 +/- 99 mL to 111 +/- 72 mL (P =.015) and 210 +/- 93 g to 152 +/- 54 g (P =.002) for younger patients, 174 +/- 115 mL to 126 +/- 43 mL (P =.17) and 233 +/- 71 g to 215 +/- 65 g (P =.19) for older patients. No significant change in EF was noted in the younger age group. However, in the older age group a significant decrease to EF <25% was found in 2 patients 1 year after surgery. Moderate autograft regurgitation was also detected in 2 patients: 1 in each age group. Pressure gradients across the autograft remained within the normal range in both age groups. Two younger patients had severe homograft stenosis with peak gradients of 100 and 62 mm Hg. The older patients did not demonstrate homograft dysfunction. CONCLUSIONS: The Ross procedure can be performed in selected older adults with aortic valve disease and provides durable valves in both aortic and pulmonic positions for at least 3 years after surgery but may result in less favorable left ventricular remodeling compared with that in the younger patients. Further follow-up will be necessary to determine the long-term outcome of the Ross procedure in this older adult patient population. PMID- 11479475 TI - A functional myeloperoxidase polymorphic variant is associated with coronary artery disease in French-Canadians. AB - BACKGROUND: One of the enzymes involved in the production of free radicals in atherosclerotic plaques is myeloperoxidase (MPO). There is a functional G/A polymorphism 463 bp upstream of the transcription start site of the enzyme with the G allele associated with a higher level of MPO expression than the A allele. Considering the potential role of MPO in the process of atherosclerosis, studying the relationship between this polymorphism and the incidence of coronary artery disease (CAD) seems reasonable. METHOD: We performed a case-control study. The case group consisted of 229 patients who had angiographically proved atherosclerotic plaques in their coronary arteries. The control group consisted of 217 individuals who did not have a history of coronary artery disease, stroke, or peripheral vascular disease. RESULTS: We found that allele A of the MPO gene was less frequent in cases with CAD. In a recessive model patients with the AA genotype had a decreased risk of CAD (odds ratio 0.138, 95% confidence interval 0.040-0.474). In a dominant model a significant protective role for AA or AG versus GG was also detected (odds ratio 0.639, 95% confidence interval 0.436 0.937). CONCLUSION: Our findings suggest that the -463 G/A polymorphism of the MPO gene influences the risk of CAD. This effect may be mediated by the effect of this polymorphism on the transcription level of the MPO gene. PMID- 11479476 TI - The dual endothelin receptor antagonist tezosentan acutely improves hemodynamic parameters in patients with advanced heart failure. AB - BACKGROUND: Endothelin-1, a potent vasoconstrictor, is elevated in congestive heart failure and is postulated to play a major role in the pathogenesis of the disease. Endothelin receptor antagonism may be a specific therapeutic approach. This study was designed to determine the effective dosage range, hemodynamic effects, and tolerability of tezosentan, an intravenous dual endothelin receptor antagonist, in patients with advanced heart failure. METHODS: This randomized, double-blind, placebo-controlled multicenter trial enrolled 38 patients with symptomatic stable heart failure (New York Heart Association class III, left ventricular ejection fraction <35%) undergoing right heart catheterization. Patients were equally randomized to a 4-hour intravenous infusion of placebo or tezosentan in ascending doses (5, 20, 50, and 100 mg over 1 hour each). Angiotensin-converting enzyme inhibitors and diuretics were withheld 24hours before the study. Hemodynamics were measured during and for 4 hours after the infusion. RESULTS: Compared with placebo, tezosentan treatment produced a significant increase in cardiac index (treatment difference 0.59 L/min/m(2), P =.0001) and decreases in pulmonary and systemic vascular resistances (P or = 65 years who were discharged alive from 18 Connecticut hospitals in 1994 and 1995 with a principal discharge diagnosis of heart failure. Outcomes 6 months after discharge, including all-cause readmission and cost, heart failure-related readmission and cost, and death, were obtained from the Medicare administrative database. A 2-stage sample selection model was used to identify the independent correlates of cost. Risk scores were calculated to identify subsets of patients at risk for generating high costs. RESULTS: On average, patients discharged with heart failure incurred costs of $2388 resulting from heart failure-related admissions and $7101 resulting from admissions from any cause during the 6 months after discharge. An average admission for heart failure cost $7174, whereas an admission resulting from any cause cost $8589. The multivariate models explained 7% of the variation in cost, although clinical characteristics such as recent heart failure admissions, kidney failure, and hypertension were significant independent correlates of increased cost. Older age and a history of stroke were independently associated with decreased cost. Patients without any of the risk factors associated with increased costs still incurred $1500 to $5000, on average, in the 6 months after discharge. CONCLUSIONS: Patients with heart failure generate substantial hospital costs in the 6 months after discharge. Given the emerging evidence for effective programs to reduce readmission, investments in interventions that produce even modest reductions in risk would be economically favorable. PMID- 11479478 TI - Increased QT dispersion does not reflect the increased regional variation of cardiac sympathetic nervous activity in hypertrophic cardiomyopathy. AB - BACKGROUND: QT dispersion (QTD) reflects regional variation of ventricular repolarization. However, the relationship between QTD and the regional variation of cardiac sympathetic nerve activity in hypertrophic cardiomyopathy (HCM) is not yet elucidated. METHODS: Cardiac sympathetic nerve activity was evaluated in 25 patients with HCM by iodine 123 metaiodobenzylguanidine (MIBG) myocardial scintigraphy. With planar MIBG imaging, heart and mediastinum ratios (H/M) at early (20 minutes) and delayed (3 hours) acquisition and the washout rate (WR) were calculated. Polar maps of left ventricular myocardium were divided into 20 segments. The SD of early uptake (EU-SD), delayed uptake (DU-SD), and WR (WR-SD) in 20 segments as indices of regional variation were calculated. QT intervals were corrected by use of the Bazett formula. RESULTS: Maximum QTc correlated positively with H/M early, WR, and left ventricular wall thickness (LVWT). Minimum QTc correlated positively with WR and LVWT. Corrected QTD (QTDc) correlated negatively with EU-SD, DU-SD, and WR-SD and positively with the interventricular septal thickness/posterior wall thickness ratio. Stepwise regression analysis revealed that the most powerful determinants for maximum QTc, minimum QTc, and QTDc were WR, LVWT, and EU-SD, respectively. CONCLUSIONS: QTD correlated negatively rather than positively with the regional variability index of cardiac sympathetic nerve activity. These results suggest that increased QTD in patients with HCM may not reflect increased heterogeneity of cardiac sympathetic nerve activity. PMID- 11479480 TI - Inhibition by combined therapy with ticlopidine and aspirin of enhanced platelet aggregation during physical exercise in patients with coronary artery disease. AB - BACKGROUND: Strenuous exercise can be a major trigger for coronary thrombosis and it enhances platelet aggregation. METHODS: We evaluated the effect of antiplatelet therapy on shear stress-induced platelet aggregation (SIPA), in addition to agonist-induced aggregation, before and immediately after ergometer exercise in patients with stable coronary artery diseases (CAD). Forty-eight patients with stable CAD were randomly distributed into 3 groups: no antiplatelet drug (patient control, n = 16), aspirin (ASA) monotherapy (n = 16), and combined therapy with ticlopidine (TIC) and ASA (n = 16). RESULTS: There were significant increases in not only adenosine phosphate (ADP)- and collagen-induced platelet aggregation but also in SIPA during exercise by the patient control group. ASA monotherapy did not attenuate the enhanced ADP-induced aggregation nor SIPA. Combined ASA + TIC therapy significantly inhibited SIPA as well as ADP-induced aggregation both before and after exercise. Significant increases in levels of plasma von Willebrand factor (vWF) occurred during exercise, and these antiplatelet therapies had no apparent effect on increased vWF levels during exercise. Exercise induced a significant increase in the plasma thrombin antithrombin III complex level with no significant changes in the level of plasmin-plasmin inhibitor complex level in all 3 groups. CONCLUSIONS: Combined therapy with ASA + TIC effectively inhibited increased platelet aggregability in response to acute exercise, with no effects on coagulant or fibrinolytic potentials in patients with CAD. The data suggest that TIC combined with ASA may be superior to ASA alone in preventing acute coronary events during exercise in patients with coronary atherosclerotic disease. PMID- 11479481 TI - The effect of pravastatin and atorvastatin on coenzyme Q10. AB - BACKGROUND: Coenzyme Q10 (CoQ10) is an antioxidant and plays an important role in the synthesis of adenosine triphosphate. Studies suggest that 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors reduce CoQ10 levels; however, no studies have directly compared HMG-CoA reductase inhibitors in a randomized crossover fashion. METHODS: Twelve healthy volunteers received either 20 mg pravastatin (P) or 10 mg atorvastatin (A) for 4 weeks in a randomized crossover fashion. There was a 4- to 8-week washout period between the 2 phases. CoQ10 levels and a lipid profile were obtained. RESULTS: There was no difference in CoQ10 levels from baseline to post-drug therapy for either P or A (0.61 +/- 0.14 vs 0.62 +/- 0.2 microg/mL and 0.65 +/- 0.22 vs 0.6 +/- 0.12 microg/mL, respectively; P >.05). There was a significant difference in low-density lipoprotein (LDL) levels from baseline to post-drug therapy for both P and A (97 +/- 21 vs 66 +/- 19 mg/dL and 102 +/- 21 vs 52 +/- 14 mg/dL, respectively; P <.01). There was no significant correlation between LDL and CoQ10. CONCLUSIONS: P and A did not decrease CoQ10 despite a significant decrease in LDL levels. These findings suggest that HMG-CoA reductase inhibitors do not significantly decrease the synthesis of circulating CoQ10 in healthy subjects. Routine supplementation of CoQ10 may not be necessary when HMG-CoA reductase inhibitor therapy is administered. PMID- 11479482 TI - Clinical assessment of clonidine in the treatment of new-onset rapid atrial fibrillation: a prospective, randomized clinical trial. AB - PURPOSE: The role of digoxin and verapamil in the control of ventricular response in rapid atrial fibrillation is well established. This study investigates how clonidine compares with these standard therapies in rate control for new-onset rapid atrial fibrillation. We set out to test the hypothesis that clonidine effectively reduces heart rate in patients with new-onset rapid atrial fibrillation. SUBJECTS AND METHODS: Forty patients were seen in the emergency department with new-onset (< or =24 hours' duration), stable, rapid atrial fibrillation. Eligible patients were randomized to receive either clonidine, digoxin, or verapamil. Changes in heart rate and blood pressure over 6 hours, as well as frequency of conversion to sinus rhythm were recorded and analyzed. RESULTS: The mean reduction in heart rate over 6 hours was 44.4 beats/min (95% confidence interval [CI] 28.4-60.4 beats/min) in the clonidine group, 52.1 beats/min (95% CI 40.8-63.4 beats/min) in the digoxin group, and 41.8 beats/min (95% CI 22.5-61.0 beats/min) in the verapamil group. Analysis of variance of the heart rate changes in the 3 groups after 6 hours was not significant (P =.55). At 6 hours, 7 of 12 clonidine patients, 8 of 15 digoxin patients, and 7 of 13 verapamil patients remained in atrial fibrillation (P =.962 on chi(2)). CONCLUSION: Clonidine controls ventricular rate in new-onset atrial fibrillation with an efficacy comparable to that of standard agents. PMID- 11479484 TI - New anticoagulants. AB - BACKGROUND: Heparin and coumarins have been in clinical use for more than 50 years. Low-molecular-weight heparins (including a heparinoid), developed 25 years ago, have been used clinically for more than a decade. METHODS: In the last 10 years, several new anticoagulants that target almost every step in the coagulation pathway have been developed. Of these, 3 direct thrombin inhibitors (hirudin, bivalirudin, and argatroban) are approved for clinical use. Four other anticoagulants (activated protein C, tissue factor pathway inhibitor, synthetic pentasaccharide, and the oral thrombin inhibitor H376/95) are being evaluated or have completed evaluation in phase III studies; various other compounds are in phase II studies. RESULTS: The mechanism of action, sites of action, and potential clinical indications of these new anticoagulants are reviewed. CONCLUSIONS: To compete effectively, the new oral anticoagulants currently under development will have to be at least as effective and safe as coumarins, oral thrombin, or Factor Xa inhibitors and require less monitoring. PMID- 11479485 TI - Pentasaccharide Org31540/SR90107A clinical trials update: lessons for practice. AB - BACKGROUND: Patients undergoing orthopedic surgery are at particularly high risk for venous thromboembolism (VTE) and as such often receive prophylactic low molecular-weight heparin (LMWH) or warfarin. However, efficacy and safety limitations of these agents have prompted a search for alternative antithrombotic drugs. METHODS: A new pentasaccharide, Org31540/SR90107A, the first of a class of synthetic and highly selective Factor Xa inhibitors, was investigated in a phase II double-blind, dose-ranging study for the prevention of deep vein thrombosis in 933 patients undergoing elective total hip replacement. The assessor-blind, comparative control was the LMWH enoxaparin. Patients were randomly assigned to 1 of 5 Org31540/SR90107A treatment groups (0.75, 1.5, 3, 6, or 8 mg/d subcutaneously) or to 30 mg enoxaparin subcutaneously every 12 hours. Treatment was started after surgery and continued for 5 to 10 days. Principal outcomes, determined on the basis of standard criteria, included venographically detected deep vein thrombosis and major bleeding. The 6.0- and 8.0-mg groups were discontinued early, in accordance with a protocol-defined safety rule. RESULTS: This phase II study showed improved dose-related efficacy and safety for Org31540/SR90107A versus enoxaparin and led to selection of 2.5 mg once daily as the optimal Org31540/SR90107A dose for large phase III trials in the prevention of VTE after total hip replacement, total knee replacement, and surgery for fractured hip. These studies have now been completed. CONCLUSIONS: Org31540/SR90107A is a new synthetic antithrombotic agent that has demonstrated a significant efficacy and safety dose effect in VTE prophylaxis after orthopedic surgery. PMID- 11479486 TI - New data in treatment of acute coronary syndromes. AB - BACKGROUND: Anticoagulants are important in the treatment of acute coronary syndromes (ACS). When given in combination with aspirin, unfractionated heparin (UHF) improves the clinical outcome in patients who have ACS without ST-segment elevation. UHF also has potential benefits when administered in conjunction with fibrinolytic agents, especially those that are fibrin-specific. The anticoagulant effect of standard heparin is unpredictable, and excessive bleeding complications have been observed in nearly all trials. METHODS: New anticoagulants, including direct antithrombins, low-molecular-weight heparins, and more recently, agents with a pure anti-Factor Xa effect, have been tested in patients with ACS and found to have varying degrees of clinical relevance. RESULTS: One new synthetic pentasaccharide, Org31540/SR90107A, an antithrombin III-binding selective inhibitor of Factor Xa, has been studied in more than 300 patients with acute myocardial infarction with ST-segment elevation (PENTALYSE trial). In an ongoing trial (PENTUA), Org31540/SR90107A is being compared with enoxaparin in patients with ACS, with and without ST-segment elevation. CONCLUSIONS: Org31540/SR90107A has shown promising results in ACS with ST-segment elevation. Efficacy in ACS without ST-segment elevation is currently under evaluation. PMID- 11479487 TI - Recent advances in anticoagulant therapy for acute coronary syndromes. AB - BACKGROUND: Despite major advances in the treatment of acute coronary syndromes (ACS), ischemic heart disease is still the leading cause of death in the industrialized world. Although unfractionated heparin (UFH) has been the anticoagulant of choice in ACS treatments, both low-molecular-weight heparins and direct thrombin inhibitors are at least as effective as UFH in comparative clinical trials. UFH has been linked to a discontinuation rebound in thrombin generation, which is associated with greater ischemic end points than those with the direct thrombin inhibitor hirudin. Furthermore, UFH can increase platelet activation in ACS. METHODS: This review summarizes the investigation of new molecular approaches to further improve clinical outcomes in ACS. RESULTS: The more recently developed synthetic pentasaccharide Org31540/SR90107A provides potent antithrombotic activity through selective inhibition of Factor Xa by high affinity binding to antithrombin III. Unlike UFH, the pentasaccharide does not bind platelet factor 4 and is not associated with platelet activation. New approaches to coronary thrombosis prevention/treatment that involve inhibition higher up the extrinsic coagulation pathway have been suggested. In view of its undesirable features, UFH may become obsolete with the advent of newer antithrombotic agents. CONCLUSIONS: Applications for the pentasaccharide and other new anticoagulants await results from definitive, large-scale trials. PMID- 11479489 TI - Statistics for the rest of us. PMID- 11479490 TI - Use of ultrafiltration during and after cardiopulmonary bypass in children. PMID- 11479491 TI - Intraoperative options for treating atrial fibrillation associated with mitral valve disease. PMID- 11479492 TI - Interpretation of nonfatal events after cardiac surgery: actual versus actuarial reporting. AB - OBJECTIVE: To describe the cumulative incidence (actual) method of analysis and to contrast it with the Kaplan-Meier method. METHOD: We use data on porcine valve replacement to illustrate these two statistical techniques. RESULTS: The "actual" analysis estimates the percentage of events expected to occur. The percentage given by the Kaplan-Meier method is much larger. CONCLUSION: Actual (cumulative incidence) analysis is preferred for estimating the probability of occurrence of a nonfatal time-related event. PMID- 11479493 TI - A prospective randomized study comparing volume-standardized modified and conventional ultrafiltration in pediatric cardiac surgery. AB - BACKGROUND: Modified ultrafiltration has been touted as superior to conventional ultrafiltration for attenuating the consequences of hemodilution after cardiac surgery with cardiopulmonary bypass in children. We conducted a prospective randomized study to test the hypothesis that modified and conventional ultrafiltration have similar clinical effects when a standardized volume of fluid is removed. METHODS: From October 1998 to September 1999, 110 children weighing 15 kg or less (median weight 6.1 kg, median age 6.3 months) undergoing surgery with cardiopulmonary bypass for functionally biventricular congenital heart disease were randomized to conventional (n = 67) or arteriovenous modified ultrafiltration (n = 43) for hemoconcentration. The volume of fluid removed with both methods was standardized as a percentage of effective fluid balance (the sum of prime volume and volume added during cardiopulmonary bypass minus urine output): in patients weighing less than 10 kg, 50% of effective fluid balance was removed, whereas 60% was removed in patients weighing 10 to 15 kg. Hematocrit, hemodynamics, ventricular function, transfusion of blood products, and postoperative resource use were compared between groups. RESULTS: There were no significant differences between groups in age, weight, or duration of cardiopulmonary bypass. The total volume of fluid added in the prime and during bypass was greater in patients undergoing conventional ultrafiltration than in those receiving modified ultrafiltration (205 +/- 123 vs 162 +/- 74 mL/kg; P =.05), although the difference was due primarily to a greater indexed priming volume in patients having conventional ultrafiltration. There was no difference in the percentage of effective fluid balance that was removed in the 2 groups. Accordingly, the volume of ultrafiltrate was greater in patients receiving conventional than modified ultrafiltration (95 +/- 63 vs 68 +/- 28 mL/kg; P =.01). Preoperative and postoperative hematocrit levels were 35.6% +/- 6.6% and 36.3% +/- 5.6% in patients having conventional ultrafiltration and 34.4% +/- 6.7% and 38.7% +/- 7.5% in those having modified ultrafiltration. By repeated-measures analysis of variance, patients receiving modified and conventional ultrafiltration did not differ with respect to hematocrit value (P =.87), mean arterial pressure (P =.85), heart rate (P =.43), or left ventricular shortening fraction (P =.21) from baseline to the postbypass measurements. There were no differences between groups in duration of mechanical ventilation, stay in the intensive care unit, or hospitalization. CONCLUSIONS: When a standardized volume of fluid is removed, hematocrit, hemodynamics, ventricular function, requirement for blood products, and postoperative resource use do not differ between pediatric patients receiving conventional and modified ultrafiltration for hemoconcentration after cardiac surgery. PMID- 11479494 TI - Early and late results of partial plication annuloplasty for congenital mitral insufficiency. AB - BACKGROUND: Partial plication annuloplasty is the main technique for congenital mitral insufficiency because this technique allows the mitral anulus to grow, in contrast to ring annuloplasty. However, this technique is not satisfactory for mitral insufficiency with some anomalies of the mitral valve apparatus. METHODS: Forty-one patients underwent partial plication annuloplasty for mitral regurgitation from July 1979 to December 1998. Mitral regurgitation associated with an atrioventricular defect, an atrioventricular discordance, and a univentricular heart was excluded from this study. RESULTS: There were no early or late deaths. In early results, partial plication annuloplasty was more effective for mitral regurgitation with abnormality of the posterior leaflet (n = 14) or normal leaflet motion (n = 8) than with abnormality of the anterior leaflet and its apparatus (n = 14) or absence of chordae (n = 4). The mean follow up period was 145.8 months. During the follow-up period, 2 patients underwent mitral valve replacement, and a third patient underwent mitral valve repair with partial plication annuloplasty after the first repair. The main cause of mitral regurgitation of 2 of the 3 patients was absence of chordae. The actuarial freedom from reoperation rate was 94.9% +/- 3.6%, 91.9% +/- 4.7%, and 91.9% +/- 4.7% at 5, 10, and 15 years after the operation, respectively. CONCLUSION: Early and long-term results of partial plication annuloplasty were acceptable for congenital mitral insufficiency with any type of malformation of the mitral valve, and results were excellent with abnormality of the posterior leaflet and its apparatus or normal leaflet motion. However, late results were suboptimal for mitral regurgitation with absence of chordae. Other techniques, such as artificial chorda replacement, should be adapted in these cases. PMID- 11479495 TI - Early and late results of operations for defects associated with corrected transposition and other anomalies with atrioventricular discordance in a pediatric population. AB - OBJECTIVES: The purpose of this study was to determine the early and late results for children having operations for defects associated with corrected transposition of the great arteries and other anomalies with atrioventricular discordance. METHODS: Data on 111 children operated on from July 1, 1971, through January 31, 1996, including clinic records, operative reports, and follow-up visits and questionnaires, were analyzed with particular reference to variables associated with early and late mortality, reoperations, ventricular function, and status of the atrioventricular valves. RESULTS: Complex associated anomalies were common and included double-outlet right ventricle (n = 43) and situs abnormalities (n = 38). Overall early mortality was 16%; for the 29 patients operated on after 1986, early mortality was 3%. Early survival was adversely affected by patch repair of ventricular septal defect and early operative interval. Follow-up of the 93 early survivors extended to 26.5 years (mean 11.4 years). Overall survival was 77% (+/-4%) at 5 years and 67% (+/-5%) at 10 years. Late survival was adversely affected by prior operations, more severe preoperative functional class, and cardiac rhythm other than sinus. Reoperation was required for 38 (41%) patients, most commonly for conduit replacement (n = 22) or repair/replacement of the systemic ventricle atrioventricular valve (n = 13). CONCLUSIONS: These results can serve as a basis for comparison with newer surgical alternatives proposed for corrected transposition of the great arteries. PMID- 11479496 TI - Continuous perfusion of pulmonary arteries during total cardiopulmonary bypass favorably affects levels of circulating adhesion molecules and lung function. AB - OBJECTIVES: Lung injury is a serious complication of cardiopulmonary bypass in infants with congenital heart disease and pulmonary hypertension. Cessation of blood flow in the pulmonary arteries during cardiopulmonary bypass is known to provoke lung dysfunction. We assessed the effect of continuous pulmonary perfusion on circulating adhesion molecules and on lung function. METHODS: Fourteen infants with congenital heart disease and pulmonary hypertension were enrolled in the study. During total cardiopulmonary bypass, 8 patients underwent continuous perfusion of the pulmonary arteries (perfusion group), and the remaining 6 patients did not (control group). Plasma levels of circulating intercellular adhesion molecule 1, soluble granule membrane protein 140, and sialyl Lewis(x) and PaO (2)/fraction of inspired oxygen ratios were measured before commencement and serially for 24 hours after termination of bypass. RESULTS: Plasma levels of circulating intercellular adhesion molecule 1 decreased significantly at the termination of bypass in both groups but returned to prebypass levels immediately in the control group, whereas in the perfusion group the values remained significantly less than those before bypass. Plasma levels of soluble granule membrane protein 140 in the control group were significantly higher at 6 and 12 hours after bypass than levels before bypass, whereas in the perfusion group the values remained at the prebypass level throughout the postbypass period. Trends of plasma levels of sialyl Lewis(x) were alike in both groups. PaO (2)/fraction of inspired oxygen ratios in the control group decreased significantly from 6 hours after bypass, whereas values in the perfusion group remained at the prebypass value throughout the postbypass period. CONCLUSIONS: This study suggests that in infants having congenital heart disease and pulmonary hypertension, continuous pulmonary perfusion during total cardiopulmonary bypass minimizes ischemic insult and neutrophil-endothelial interaction mediated by adhesion molecules in the pulmonary microvessels. PMID- 11479497 TI - Radiofrequency modified maze in patients with atrial fibrillation undergoing concomitant cardiac surgery. AB - OBJECTIVE: In the majority of patients with chronic atrial fibrillation the arrhythmia will persist after correction of the underlying structural abnormality. The maze procedure is an effective surgical method to eliminate atrial fibrillation and to restore atrial contractility. METHODS: In this study we used radiofrequency energy to create lines of conduction block in both atria during cardiac surgery as a modification of the maze III procedure. One hundred twenty-two patients with atrial fibrillation for at least 1 year and structural heart disease underwent open heart operation and a radiofrequency modified maze procedure. RESULTS: In 108 (89%) of 122 patients mitral valve surgery was performed, and in this group 86 patients (80%) underwent 121 concomitant procedures. Fourteen patients (11%) underwent cardiac surgery not involving the mitral valve. The additional crossclamp time required for the left atrial part of the radiofrequency modified maze procedure was 14 +/- 3 minutes. The in-hospital mortality rate was 4.1%. The overall 39-month survival was 90%, and freedom of atrial flutter or atrial fibrillation was 78.5% +/- 5.1%. Eighty-nine survivors with sinus, atrial rhythm, or atrioventricular sequential pacemaker had Doppler echocardiography, and right atrial transport function was documented in 83% and left atrial transport function in 77% of patients. CONCLUSION: We concluded that the radiofrequency modified maze procedure as an adjunctive procedure is safe, time-sparing, and effective in eliminating atrial fibrillation and restoring atrial transport function. PMID- 11479498 TI - Twenty-year comparison of tissue and mechanical valve replacement. AB - OBJECTIVE: We sought to compare outcomes with tissue and St Jude Medical mechanical valves over a 20-year period. METHODS: Valve-related events and overall survival were analyzed in 2533 patients 18 years of age or older undergoing initial aortic, mitral, or combined aortic and mitral (double) valve replacement with a tissue valve (Hancock, Carpentier-Edwards porcine, or Carpentier-Edwards pericardial) or a St Jude Medical mechanical valve. Total follow-up was 13,390 patient-years. There were 666 St Jude Medical aortic valve replacements, 723 tissue aortic valve replacements, 513 St Jude Medical mitral valve replacements, 402 tissue mitral valve replacements, 161 St Jude Medical double valve replacements, and 68 tissue double valve replacements. The mean age was 68 +/- 13.3 years (St Jude Medical valve, 64.5 +/- 12.9; tissue valve, 72.0 +/- 12.6). RESULTS: There were no overall differences in survival between tissue and mechanical valves. Multivariable analysis indicated that the type of valve did not affect survival. Analysis by age less than 65 years or 65 years or older and presence or absence of coronary disease revealed similar long-term survival in all subgroups. The risk of hemorrhage was lower in patients receiving tissue aortic valve replacements but was not significantly different in patients receiving mitral valve or double valve replacements. Thromboembolism rates were similar for tissue and mechanical valve recipients. However, reoperation rates were significantly higher in patients receiving both aortic and mitral tissue valves. The reoperation hazard increased progressively with time both in patients receiving aortic and in those receiving mitral tissue valves. Overall valve complications were initially higher with mechanical aortic valves but not with mechanical mitral valves. However, valve complication rates later crossed over, with higher rates in tissue valve recipients after 7 years in patients undergoing mitral valve replacement and 10 years in those undergoing aortic valve replacement. CONCLUSIONS: Tissue and mechanical valve recipients have similar survival over 20 years of follow-up. The primary tradeoff is an increased risk of hemorrhage in patients receiving mechanical aortic valve replacements and an increased risk of late reoperation in all patients receiving tissue valve replacements. The risk of tissue valve reoperation increases progressively with time. PMID- 11479499 TI - Valve-preserving aortic replacement: does the additional repair of leaflet prolapse adversely affect the results? AB - OBJECTIVE: Valve-preserving aortic replacement has evolved into an accepted therapeutic option for aortic ectasia with morphologically intact leaflets. Some patients, however, exhibit additional leaflet prolapse. We compared the results of established valve-preserving techniques with those of the combination of valve preserving aortic surgery and additional repair of leaflet prolapse. METHODS: Between October 1995 and March 2000, 99 patients underwent valve-preserving root replacement by means of root remodeling or valve reimplantation for acute dissection (n = 25), chronic dissection (n = 4), or aneurysm (n = 70). In group A (63 patients) either root remodeling (n = 49) or valve reimplantation (n = 14) was performed with a standard technique. In group B (36 patients) valvepreserving aortic replacement (remodeling, n = 31; reimplantation, n = 5) was combined with repair of leaflet prolapse in the presence of bicuspid (n = 24) or tricuspid (n = 12) valve anatomy. Additional replacement of the aortic arch was required more frequently in group A (group A, n = 43; group B, n = 14; P =.006); otherwise, the groups were comparable. RESULTS: Cardiopulmonary bypass (group A, 133 +/- 31 minutes; group B, 117 +/- 30 minutes; P =.006) and myocardial ischemia times (group A, 96 +/- 25 minutes; group B, 88 +/- 20 minutes; P =.05) were significantly longer in group A. Mortality was not significantly different between groups (group A, 4.8%; group B, 0%). One patient in each group underwent secondary valve replacement, and all other patients had stable valve function. Freedom from aortic regurgitation of grade 2 or greater after 48 months was 93.0% in both groups. CONCLUSION: Repair of leaflet prolapse in conjunction with valve preserving root replacement leads to midterm results that are equal to those of valve-preserving root replacement for morphologically intact leaflets. PMID- 11479500 TI - Risk factors for early hospital readmission after cardiac operations. AB - OBJECTIVE: Early hospital readmissions after cardiac procedures are both costly and harmful to patients. We investigated the factors that predispose to readmission to develop strategies to minimize this problem. METHODS: As part of a prospective data collection, patients having cardiac procedures at our institution are routinely tracked for 30 days after their discharge from the hospital. We reviewed 2650 patients in our cardiac database who underwent operations over the past 5 years. We used univariate and multivariate statistical techniques to identify risks for readmission. RESULTS: Of 2574 discharged patients, 252 (9.8%) required readmission. The most common causes of readmission are cardiac (42%), pulmonary (19%), gastrointestinal (10%), extremity complications (6.7%; deep vein thrombophlebitis, peripheral arterial vascular disease, and saphenous vein harvest site problems), sternal wound problems (7.5%), and metabolic problems (4%). Of more than 70 variables studied, only 6 are significant multivariate predictors of readmission: female sex (P =.002); diabetes (P =.001); chronic lung problems (P =.011); increased distance between home and hospital (P >.001); preoperative atrial fibrillation (P =.002); and preoperative chronic renal insufficiency (P =.002). Type of operation, redo procedures, and other intraoperative and postoperative variables are not important multivariate predictors of readmission. Prolonged hospital length of stay for the initial procedure did not cause more frequent readmission. The costs of initial hospitalization (operating room costs combined with postoperative in hospital costs) were not significantly increased in those patients who required readmission. CONCLUSIONS: The high-risk patient for readmission is a woman with diabetes, chronic lung disease, renal insufficiency, and preoperative atrial fibrillation who lives at a distance from the hospital. Readmission does not depend on periprocedural variables (eg, cardiopulmonary bypass time) or on postoperative complications. High procedural costs from the initial hospitalization do not predispose to readmission. These results suggest interventions that may reduce readmission. PMID- 11479501 TI - Assessment of effective orifice area of prosthetic aortic valves with Doppler echocardiography: an in vivo and in vitro study. AB - OBJECTIVES: We sought to evaluate the Doppler assessment of effective orifice area in aortic prosthetic valves. The effective orifice area is a less flow dependent parameter than Doppler gradients that is used to assess prosthetic valve function. However, in vivo reference values show a pronounced spread of effective orifice area and smaller orifices than expected compared with the geometric area. METHODS: Using Doppler echocardiography, we studied patients who received a bileaflet St Jude Medical valve (n = 75; St Jude Medical, Inc, St Paul, Minn) or a tilting disc Omnicarbon valve (n = 46; MedicalCV, Incorporated, Inver Grove Heights, Minn). The prosthetic valves were also investigated in vitro in a steady-flow model with Doppler and catheter measurements in the different orifices. The effective orifice area was calculated according to the continuity equation. RESULTS: In vivo, there was a wide distribution with the coefficient of variation (SD/mean x 100%) for different valve sizes ranging from 21% to 39% in the St Jude Medical valve and from 25% to 33% in the Omnicarbon valve. The differences between geometric orifice area and effective orifice area in vitro were 1.26 +/- 0.41 cm(2) for St Jude Medical and 1.17 +/- 0.38 cm(2) for Omnicarbon valves. The overall effective orifice areas and peak catheter gradients were similar: 1.35 +/- 0.37 cm(2) and 25.9 +/- 16.1 mm Hg for St Jude Medical and 1.46 +/- 0.49 cm(2) and 24.6 +/- 17.7 mm Hg for Omnicarbon. However, in St Jude Medical valves, more pressure was recovered downstream, 11.6 +/- 6.3 mm Hg versus 3.4 +/- 1.6 mm Hg in Omnicarbon valves (P =.0001). CONCLUSIONS: In the patients, we found a pronounced spread of effective orifice areas, which can be explained by measurement errors or true biologic variations. The in vitro effective orifice area was small compared with the geometric orifice area, and we suspect that nonuniformity in the spatial velocity profile causes underestimation. The St Jude Medical and Omnicarbon valves showed similar peak catheter gradients and effective orifice areas in vitro, but more pressure was recovered in the St Jude Medical valve. The effective orifice area can therefore be misleading in the assessment of prosthetic valve performance when bileaflet and tilting disc valves are compared. PMID- 11479502 TI - Complementary saphenous grafting: long-term follow-up. AB - OBJECTIVE: Recently we reviewed the 10-year clinical and angiographic outcomes of sequential internal thoracic artery grafting. Most of the patients also received complementary saphenous grafts, and their overall long-term patency rates were surprisingly high. Therefore, we decided to analyze these results in more detail. METHODS: The first consecutive 500 patients having received at least one sequential internal thoracic artery graft between October 1985 and August 1991 were restudied retrospectively. The saphenous grafts were only used to achieve complete revascularization in addition to complex arterial grafting on less significant or remote coronary vessels. A total of 161 patients consented to a late angiographic restudy at a mean postoperative interval of 7.5 years (1-12.2 years). RESULTS: At 5 and 10 years postoperatively, freedom from angina was 96% and 82%, and freedom from any cardiac event was 92.8% and 69%, respectively. Only 15 (3.1%) patients needed additional revascularization (0.3% per patient-year): 4 coronary artery bypass grafting (0.8%) and 11 percutaneous transluminal coronary angioplasty (2.3%). The overall patency and intactness rates of saphenous anastomoses were 72.5% and 60.2%, respectively. There was a significant difference between the patency and intactness of sequential versus single anastomoses: 76% versus 60% and 64.5% versus 44.4%, respectively. There was no significant difference in either patency or intactness between right internal thoracic and sequential saphenous grafts anastomosed to the right coronary artery: 83.4% versus 75.2% and 77.8% versus 62.4%, respectively. The same was true for the anastomoses to the "remote area" (distal circumflex, distal right coronary artery). CONCLUSIONS: Complementary sequential saphenous grafting still deserves consideration in some patients below 70 years of age, particularly for those with disease in the "remote area": the distal circumflex and right coronary branches. PMID- 11479503 TI - The human internal thoracic artery releases more nitric oxide in response to vascular endothelial growth factor than the human saphenous vein. AB - OBJECTIVE: Endothelial nitric oxide inhibits smooth muscle cell proliferation, reducing the chance of vascular intimal thickening. In this study we investigated whether the superior long-term patency of the internal thoracic artery in human coronary bypass grafting compared with that of the saphenous vein could be explained by different levels of nitric oxide production. METHODS: The baseline endogenous nitric oxide production appeared to be 50% higher in the internal thoracic artery than in the saphenous vein. Previously, it was shown that vascular endothelial growth factor and the vascular endothelial growth factor receptors KDR (Flk-1) and Flt-1 are expressed in both internal thoracic arteries and saphenous veins and that vascular endothelial growth factor receptor density was higher in internal thoracic arteries than in saphenous veins. Therefore, we also investigated the influence of vascular endothelial growth factor on nitric oxide release in both the internal thoracic artery and the saphenous vein. RESULTS: Vascular endothelial growth factor augmented nitric oxide production by approximately 50% in the saphenous vein and 100% in the internal thoracic artery. As shown by means of immunohistochemistry, expression of endothelial constitutive nitric oxide synthase was similar in the internal thoracic artery and the saphenous vein, and no inducible nitric oxide synthase was expressed in any of the vascular segments. CONCLUSION: Vascular endothelial growth factor augments endothelial constitutive nitric oxide synthase-dependent nitric oxide release to a greater extent in the internal thoracic artery than in the saphenous vein. These findings may help to explain the long-term superiority of the internal thoracic artery versus the saphenous vein as a conduit for coronary artery bypass. PMID- 11479504 TI - Prevention of allograft heart valve failure in a rat model. AB - OBJECTIVE: Allograft heart valves are commonly used in cardiac surgery. Despite mounting evidence that these valves are immunogenic, leading to premature failure, current clinical practice does not attempt to minimize or control such a response. The objective of this study was to evaluate immune modulatory approaches to ameliorate allograft valve failure in a rat model. METHOD: Aortic valve grafts were implanted infrarenally into Lewis rat recipients (n = 32). There were 4 transplant groups: syngeneic grafts (Lewis to Lewis), untreated allografts (Brown Norway to Lewis), allograft recipients treated with cyclosporine (INN: ciclosporin) (10 mg/kg per day for 7 or 28 days), and allograft recipients treated with anti-alpha4 integrin and anti-beta2 integrin monoclonal antibodies for 7 days. At 7 and 28 days the valves were examined for structural integrity and cellular infiltration. RESULTS: Both cyclosporine and anti-alpha4/beta2 integrin treatment resulted in significant reduction in leaflet infiltration by macrophages (ED1(+)), T cells (CD3(+)), and CD8(+) T cells at 7 days with preservation of structural integrity when compared with control allografts. Twenty-eight days after implantation, daily treatment with cyclosporine preserved leaflet structural integrity and inhibited cellular infiltration. However, a short course of cyclosporine (7 days) failed to prevent destruction of the valves at 28 days. CONCLUSIONS: Immune modulatory approaches aimed at T-cell activation or trafficking decrease leaflet cellular infiltration and prevent allograft valve structural failure. However, short-course therapy does not appear to be sufficient and must be maintained to allow long-term preservation of leaflet structural integrity (28 days). PMID- 11479505 TI - Fast-tracking pulmonary resections. AB - OBJECTIVE: We streamlined our care after pulmonary resection for quality and cost effectiveness. METHODS: A single surgeon performed 500 consecutive pulmonary resections through a thoracotomy over a 2(3/4)-year period in a university setting. Patients were extubated in the operating room and sent directly to their hospital room. Chest tubes were placed to water seal and removed on postoperative day 2 if there was no air leak and drainage was less then 400 mL/d. Epidural catheters were used and removed by postoperative day 2. The plan for each day and discharge on postoperative day 3 or 4 was reviewed with the patients and families daily during rounds. The patient went home the day the last chest tube was removed. Persistent air leaks were treated with Heimlich valves. RESULTS: There were 500 patients (338 men), with a median age of 58 years (range, 3-87 years). Of these patients, 293 had pre-existing conditions. Seventy-three (15%) patients had been denied operations by at least one other surgeon. Four hundred nineteen (84%) patients had successful placement of a functioning preoperative epidural catheter. Pneumonectomy was performed in 32 (6%) patients, segmentectomy was performed in 16 (3%) patients, and lobectomy, sleeve lobectomy, and/or bilobectomy was performed in 194 (39%) patients. Nonanatomic resections were performed for metastasectomy. This included a single wedge resection in 161 (32%) patients and multiple wedge resections in 97 (19%) patients. A total of 482 (96%) patients were extubated in the operating room, and 380 (76%) patients were sent to their hospital room. The remaining 120 patients went to the intensive care unit for a median of 1 day (range, 1-41 days). Complications occurred in 107 (21%) patients, and operative mortality was 2.0%. Median day of discharge was postoperative day 4 (range, 2-119 days). A total of 327 (65%) patients left the hospital on postoperative day 4 or sooner. By survey, 97% of patients had excellent or good satisfaction with their care at hospital discharge, and 91% were extremely happy or satisfied at the 2-week follow-up contact. CONCLUSIONS: Most patients who undergo elective pulmonary resection can be extubated immediately after the operation, go directly to their room and avoid the intensive care unit, be discharged on postoperative day 3 or 4, and have minimal morbidity and mortality with high satisfaction both at discharge and at the 2 week follow-up contact. Techniques that seem to accomplish this include the following: the use of a water seal, removal of epidural catheters on postoperative day 2, early chest tube management, treatment of persistent air leaks with Heimlich valves, and daily reinforcement of the planned events for each day, as well as on the date of discharge with the patients and their families. PMID- 11479506 TI - Pathologic N0 status in pulmonary adenocarcinoma is predictable by combining serum carcinoembryonic antigen level and computed tomographic findings. AB - OBJECTIVES: It is not clear whether lymphadenectomy has therapeutic benefit in non-small cell lung cancer management. To avoid unnecessary lymphadenectomy, we attempted to identify clinical or radiologic predictors of pathologic N0 disease in patients with peripheral adenocarcinoma. METHODS: From August 1992 through April 1997, 269 consecutive patients with peripheral adenocarcinoma who underwent major lung resection and systematic lymph node dissection were enrolled in this study. We reviewed their contrast-enhancement computed tomographic scans and recorded the maximum dimension of tumors both on pulmonary (pDmax) and on mediastinal (mDmax) window setting images, the largest dimension perpendicular to the maximum axis on both pulmonary (pDperp) and mediastinal (mDperp) window setting images, and the size of all detectable hilar-mediastinal lymph nodes. We defined a new radiologic parameter, tumor shadow disappearance rate (TDR), which is calculated with the following formula: TDR = 1 - (mDmax x mDperp)/(pDmax x pDperp). RESULTS: In multivariable analysis a lower serum carcinoembryonic antigen level and a higher tumor shadow disappearance rate were significant predictors of pathologic N0 disease. Lymph node size on computed tomographic scanning was not a significant predictor. Among 59 patients with a normal preoperative carcinoembryonic antigen level and a tumor shadow disappearance rate of 0.8 or more, 58 (98%) patients had pathologic N0 disease, and the other patient had pathologic N1 disease. CONCLUSIONS: Mediastinal lymph node involvement was not found in patients with a normal preoperative serum carcinoembryonic antigen level and a tumor shadow disappearance rate 0.8 or more. The patients who meet these criteria may be successfully managed with major lung resection without systematic mediastinal lymphadenectomy. PMID- 11479507 TI - Retrograde cerebral perfusion provides negligible flow through brain capillaries in the pig. AB - OBJECTIVES: Although retrograde cerebral perfusion is being used clinically during aortic arch surgery, whether retrograde flow perfuses the brain effectively is still uncertain. METHODS: Fourteen pigs were cooled to 20 degrees C with cardiopulmonary bypass and perfused retrogradely via the superior vena cava for 30 minutes: 7 underwent standard retrograde cerebral perfusion and 7 underwent retrograde perfusion with occlusion of the inferior vena cava. Antegrade and retrograde cerebral blood flow were calculated by quantitating fluorescent microspheres trapped in brain tissue after the animals were put to death; microspheres returning to the aortic arch, the inferior vena cava, and the descending aorta were also analyzed during retrograde cerebral perfusion. RESULTS: Antegrade cerebral blood flow was 16 +/- 7.7 mL. min(-1). 100 g(-1) before retrograde cerebral perfusion and 22 +/- 6.3 mL. min(-1). 100 g(-1) before perfusion with caval occlusion (P =.14). During retrograde perfusion, calculations based on the number of microspheres trapped in the brain showed negligible flows (0.02 +/- 0.02 mL. min(-1). 100 g(-1) with retrograde cerebral perfusion and 0.04 +/- 0.02 mL. min(-1). 100 g(-1) with perfusion with caval occlusion; P =.09): only 0.01% and 0.02% of superior vena caval inflow, respectively. Less than 13% of retrograde superior vena caval inflow blood returned to the aortic arch with either technique. During retrograde cerebral perfusion, more than 90% of superior vena caval input was shunted to the inferior vena cava and was then recirculated, as indicated by rapid development of an equilibrium in microspheres between the superior and inferior venae cavae. With retrograde perfusion and inferior vena caval occlusion, less than 12% of inflow returned to the descending aorta and only 0.01% of microspheres. CONCLUSIONS: The paucity of microspheres trapped within the brain indicates that retrograde cerebral perfusion, either alone or combined with inferior vena caval occlusion, does not provide sufficient cerebral capillary perfusion to confer any metabolic benefit. The slightly improved outcome previously reported with retrograde cerebral perfusion during prolonged circulatory arrest in this model may be a consequence of enhanced cooling resulting from perfusion of nonbrain capillaries and from venoarterial and venovenous shunting. PMID- 11479508 TI - Prediction of safe duration of hypothermic circulatory arrest by near-infrared spectroscopy. AB - OBJECTIVE: Hypothermic circulatory arrest is widely used for adults with aortic arch disease as well as for children with congenital heart disease. At present, no method exists for monitoring safe duration of circulatory arrest. Near infrared spectroscopy is a new technique for noninvasive monitoring of cerebral oxygenation and energy state. In the current study, the relationship between near infrared spectroscopy data and neurologic outcome was evaluated in a survival piglet model with hypothermic circulatory arrest. METHODS: Thirty-six piglets (9.36 +/- 0.16 kg) underwent circulatory arrest under varying conditions with continuous monitoring by near-infrared spectroscopy (temperature 15 degrees C or 25 degrees C, hematocrit value 20% or 30%, circulatory arrest time 60, 80, or 100 minutes). Each setting included 3 animals. Neurologic recovery was evaluated daily by neurologic deficit score and overall performance category. Brain was fixed in situ on postoperative day 4 and examined by histologic score. RESULTS: Oxygenated hemoglobin signal declined to a plateau (nadir) during circulatory arrest. Time to nadir was significantly shorter with lower hematocrit value (P <.001) and higher temperature (P <.01). Duration from reaching nadir until reperfusion ("oxygenated hemoglobin signal nadir time") was significantly related to histologic score (r (s) = 0.826), neurologic deficit score (r (s) = 0.717 on postoperative day 1; 0.716 on postoperative day 4), and overall performance category (r (s) = 0.642 on postoperative day 1; 0.702 on postoperative day 4) (P <.001). All animals in which oxygenated hemoglobin signal nadir time was less than 25 minutes were free of behavioral or histologic evidence of brain injury. CONCLUSION: Oxygenated hemoglobin signal nadir time determined by near-infrared spectroscopy monitoring is a useful predictor of safe duration of circulatory arrest. Safe duration of hypothermic circulatory arrest is strongly influenced by perfusate hematocrit value and temperature during circulatory arrest. PMID- 11479509 TI - Establishment of a local cooling model against spinal cord ischemia representing prolonged induction of heat shock protein. AB - OBJECTIVES: Paraplegia is one of the serious complications of thoracoabdominal aortic operations. Regional hypothermia protects against spinal cord ischemia although the protective mechanism remains unknown. We attempted to create a simple model of local cooling under transient spinal cord ischemia and evaluated the effect using functional and histologic findings. METHODS: Male domesticated rabbits were divided into 3 groups: control, normothermic group (group N), and local hypothermic group (group H). A balloon catheter was used for spinal cord ischemia by abdominal aortic clamping. A cold pack attached to the lumbar region could lower the regional cord temperature initially. Neurologic function was evaluated by the Johnson score. Cell damage was analyzed by observing motor neurons with the use of hematoxylin and eosin staining, terminal deoxynucleotidyl transferase-mediated deoxy-uracil triphosphate biotin in situ nick end labeling (TUNEL), and immunoreactivity of heat shock protein. RESULTS: Physiologic estimation showed that local hypothermia improved the functional deficits (group N, 1.3 +/- 0.9; group H, 4.9 +/- 0.3; P =.0020). Seven days after reperfusion, there was a significant difference in the motor neuron numbers between groups N and H (group N, 7.2 +/- 1.9; group H, 20.4 +/- 3.2; P =.0090). The number of TUNEL-positive motor neurons was reduced significantly (group N, 7.2 +/- 2.4; group H, 1.0 +/- 0.7; P =.0082). Heat shock protein immunoreactivity was prolonged up to 2 days after reperfusion in the hypothermic group. CONCLUSIONS: These results suggest that local hypothermia extended the production of heat shock protein in spinal cord motor neurons after reperfusion and inhibited their apoptotic change. PMID- 11479510 TI - Endothelin-1 during and after cardiopulmonary bypass: association to graft sensitivity and postoperative recovery. AB - OBJECTIVE: Our objectives are 2-fold: (1) to serially measure the release of endothelin and graft-conduit endothelin sensitivity during and after coronary artery bypass grafting and (2) to define potential relationships of changes in endothelin levels to perioperative parameters. METHODS: Endothelin plasma content was measured in patients (n = 105) undergoing bypass grafting from select vascular compartments before operations and at specific intervals up to 24 hours postoperatively. Endothelin sensitivity was determined in isolated internal thoracic artery segments. RESULTS: Systemic arterial and pulmonary arterial endothelin levels were increased by approximately 50% immediately after bypass grafting and increased by another 85% during the first 24 hours postoperatively. Endothelin levels were highest in patients with prolonged ventilatory requirements and extended stays in the intensive care unit (10.2 +/- 0.8 vs 13.2 +/- 1.1 fmol/mL, P =.02, and 9.8 +/- 0.7 vs 13.9 +/- 1.2 fmol/mL, P =.01, respectively. Endothelin sensitivity of the internal thoracic artery was increased in patients requiring prolonged vasodilator support with nitroglycerin. CONCLUSIONS: Systemic and pulmonary arterial endothelin levels remained increased for at least 24 hours postoperatively. Prolonged pharmacologic management and increased intensive care unit stay were associated with increased systemic endothelin release and heightened graft-conduit sensitivity to endothelin. PMID- 11479511 TI - Endothelin receptor subtype A blockade selectively reduces pulmonary pressure after cardiopulmonary bypass. AB - BACKGROUND: The bioactive peptide endothelin-1 is elevated during and after cardiopulmonary bypass and exerts cardiovascular effects through its 2 receptor subtypes, endothelin-1A and endothelin-1B. Increased endothelin-1A receptor stimulation after cardiopulmonary bypass can cause increased pulmonary vascular resistance and modulate myocardial contractility. However, whether and to what degree selective endothelin-1A blockade influences these parameters in the postbypass setting is not completely understood. OBJECTIVES: Our objective was to measure left ventricular function and hemodynamics in a porcine model of cardiopulmonary bypass after selective blockade of endothelin-1A. METHODS: Adult pigs (n = 23) underwent 90 minutes of cardiopulmonary bypass and were randomized 30 minutes after bypass to receive a selective endothelin-1A antagonist (TBC 11251, 10 mg/kg; n = 13) or saline vehicle (n = 10). RESULTS: After bypass and before randomization, pulmonary vascular resistance rose nearly 4-fold, and left ventricular preload recruitable stroke work fell to one third of baseline values (both P <.05). In the vehicle group pulmonary vascular resistance continued to rise, and preload recruitable stroke work remained reduced. However, after endothelin-1A blockade, the rise in pulmonary vascular resistance was significantly blunted compared with that in the vehicle group. Moreover, the reduction in pulmonary vascular resistance with endothelin-1A blockade was achieved without a significant change in systemic perfusion pressures. CONCLUSIONS: The present study demonstrated that increased activity of the endothelin-1A receptor likely contributes to alterations in pulmonary vascular resistance in the postbypass setting. Selective endothelin-1A blockade may provide a means to selectively decrease pulmonary vascular resistance without significant effects on systemic hemodynamics. PMID- 11479512 TI - Hemodynamic effects of S-nitrosocysteine, an intravenous regional vasodilator. AB - BACKGROUND: S-nitrosocysteine is a carrier form of nitric oxide that can be delivered intravenously. S-nitrosocysteine is rapidly metabolized by plasma (half life = 2-3 seconds), forming nitric oxide and cysteine. With its short half-life and potent vasodilatory properties, S-nitrosocysteine may be useful as a pulmonary vasodilating agent in cases of postoperative and chronic pulmonary hypertension. OBJECTIVE: Our objective was to determine the hemodynamic properties of S-nitrosocysteine on the pulmonary and systemic circulations to assess its potential utility as a pulmonary vasodilatory agent. METHODS: Eleven adult swine were anesthetized. Thermodilution (Swan-Ganz; Baxter International, Inc, Deerfield, Ill) and arterial catheters were inserted. Flow probes were placed around the coronary, renal, superior mesenteric, and iliac arteries. Incremental infusion doses of S-nitrosocysteine (5-80 nmol. kg(-1). min(-1)) were delivered into the right atrium. Cardiac output, right and left heart pressures, heart rate, Pao(2), and iliac, renal, coronary, and mesenteric blood flow rates were recorded at baseline and at each infusion dose of S-nitrosocysteine. RESULTS: Low-dose S-nitrosocysteine infusion decreased mean pulmonary artery pressure (15%, P =.013) without a significant reduction in mean systemic artery pressure. Higher dose infusions produced further dose-dependent declines in pulmonary vascular resistance and measurable reductions in systemic vascular resistance (P =.01). At an S-nitrosocysteine dosage of 40 nmol. kg(-1). min(-1), there was a significant reduction in renal (P <.001) and mesenteric (P =.003) blood flow but no change in iliac (P >.2) or coronary (P >.2) blood flow. Cardiac output remained constant up to infusion rates of 40 nmol. kg(-1). min(-1) (P >.2). Doses higher than 5 nmol. kg(-1). min(-1) resulted in a substantial dose dependent reduction in Pao(2) (P <.001), suggesting dilation of atelectatic areas of the lung. CONCLUSION: S-nitrosocysteine is a potent vasodilatory agent capable of overcoming the hypoxic vasoconstrictive response of the lung. Our results suggest it may prove useful as a pulmonary vasodilatory agent at low doses. Higher dose infusions reduce mean systemic pressure and lead to compensatory reductions in renal and mesenteric blood flow without a decrease in cardiac output. PMID- 11479513 TI - Mitral valve replacement with the pulmonary autograft: the Ross II procedure. PMID- 11479514 TI - Insufficiency of the native aortic valve and left ventricular assist system inflow valve after support with an implantable left ventricular assist system: signs, symptoms, and concerns. PMID- 11479515 TI - Interventricular septal enlargement for left atrioventricular valve replacement with a larger prosthesis. PMID- 11479516 TI - Surgical approach by cervicosternolaparotomy for the treatment of extended cervical stenoses after reconstruction for caustic injury. PMID- 11479517 TI - Fistulectomy as an alternative to segmentectomy for pulmonary arteriovenous fistula. PMID- 11479518 TI - Spatial orientation of the ventricular muscle band: physiologic contribution and surgical implications. PMID- 11479519 TI - A new surgical technique for one-stage repair of interrupted aortic arch with valvular aortic stenosis. PMID- 11479520 TI - Sarcomatous mesothelioma in the left ventricle: a rare entity. PMID- 11479521 TI - Resection of ascending aortic aneurysm without use of an interposition aortic graft. PMID- 11479522 TI - Prevalence and prediction of calcification and plaques in radial artery grafts by ultrasound. PMID- 11479523 TI - Third nerve palsy after coronary artery bypass surgery. PMID- 11479524 TI - Use of pH-stat strategy for hypothermic circulatory arrest might obviate the need for leukocyte filtration. PMID- 11479525 TI - Tracheal resection associated with slide tracheoplasty for long-segment congenital tracheal stenosis involving the carina. PMID- 11479526 TI - Effect of flow competition on internal thoracic artery: postoperative velocimetric and angiographic study. PMID- 11479527 TI - Prevention of antegrade flow into the false lumen in acute aortic dissection. PMID- 11479528 TI - Aprotinin administration in the pericardial cavity does not prevent platelet activation. PMID- 11479529 TI - [Fungal allergy -clinical aspect-]. AB - Fungus is known to be one of the important causative allergens of allergic diseases such as bronchial asthma, allergic rhinitis and atopic dermatitis. Not only outdoor airborne fungi but also indoor fungi such as Aspergillus restrictus and Eurotium have recently received attention as causative fungal allergens of allergic diseases. In addition to careful history taking, in vivo testing methods such as skin test and conjunctival or bronchial provocation tests are sometimes necessary to diagnose the causative allergen. Problems requiring resolution are: 1. fungal allergens composed of multiple allergenic components, 2. difficulty of environmental monitoring of fungal allergens, 3. special characteristics as microorganisms, and 4. characteristics of normal flora allergy, etc. PMID- 11479530 TI - [Fungal allergy - fungal ecology in dwelling environments]. AB - Fungi related to allergies are commonly found in dwelling environments. The predominant fungi Cladosporium, Penicillium, Aspergillus, Alternaria, Wallemia and Rhodotorula live mainly in indoor air, house dust (HD), futons, clothes and contaminated building materials. Fungi in HD are especially important allergens. The fungal CFU and predominant fungi in HD are 10(4) - 10(6)/g and are composed of xerophilic or osmophilic species Aspergillus restrictus, Wallemia and Eurotium but not many yeasts and actinomycetes. Fungal contamination of materials is a serious human health problem because the fungal cells scatter from the materials in the air or HD. The biological activities by fungi also have health implications from the viewpoint of fungal allergens. In this paper, fungal germination, enzyme activities, contaminating cell form and viable or nonviable cells are also discussed. PMID- 11479531 TI - [A case of chromomycosis arising in a Japanese-Brazilian patient]. AB - A 41-year-old Japanese-Brazilian man, living in Japan since 1991, visited our hospital in August 1998 complaining of a scaly annular erythema which had been present on his right forearm since 1996. Granulomatous inflammation was revealed in the dermis upon skin biopsy. Sclerotic cells were present within the granulomatous lesions. Fonsecaea pedrosoi was isolated from tissue cultures. The restriction fragment length polymorphism pattern of mitochondrial DNA of the causative fungus was compatible with F. pedrosoi type 1, the commonest type in Japan. Since type 4 is usually found in South America, the patient was assumed to have become infected in Japan. An oral dose of terbinafine at 125 mg/day was ineffective. 125 mg/day single dose of terbinafine reaches an average plasma concentration of only 0.69 g/ml, where as 250 mg/day reaches 0.96 micro g/ml. Because the minimum inhibitory concentration (MIC) of terbinafine against the isolated fungus was found to be 0.76 micro g/ml, the terbinafine dose was increased from 125 mg/day to 250 mg/day, which slowly led to remission. Chromomycosis is caused by several species of dematiaceous fungi, and terbinafine efficacy may vary depending on the causative fungus. MIC values may differ even within the same species. It is important to isolate the pathogenic fungus in each case and measure the MIC value to determine the optimal dose of terbinafine. PMID- 11479532 TI - [Assimilation test of Malassezia furfur isolated from the environment]. AB - The lipophilic yeasts Malassezia species are the causative agents of tinea versicolor and known also to be a member of normal skin flora. They are commonly isolated from the skin of humans and animals, but not from the environment. This is the first report of the isolation of Malassezia sp. from the environment (a hospital floor). The results of assimilation tests of lipids and karyotyping showed that these isolates were M. furfur. They assimilated not only lipids including floor wax and car wax but also some ointments (except antifungal agents) used clinically. The results suggest that we need to take care when using such ointments to treat skin diseases. PMID- 11479533 TI - Atypical Cryptococcus neoformans isolate from an HIV-infected patient in Brazil. AB - Cryptococcus neoformans is an important fungal pathogen in immunocompromised hosts. Capsulation, urease and melanin synthesis activity of the fungus are well known virulence factors. Although artificial melanin-deficient mutants of Cr. neoformans have been investigated, the clinical mutant is rare. We found a Cr. neoformans isolate in the cerebrospinal fluid of an AIDS patient which produced a light tan colony on a caffeic acid cornmeal agar (CACA) plate. The mycological feature of the isolate was as follows; normal capsulation, defective inositol assimilation ability, serotype A; urease-positive; mating type alfa; haploid; extremely slow growth in RPMI 1640 medium, Sabouraud dextrose broth, brain heart infusion broth and yeast nitrogen base; lower production of melanin with L-DOPA substrate; and low virulence to ddY mice. We also investigated the partial DNA sequence of CNLAC1 gene between the 3085th to 3623rd base. There were many substitutions, 3 insertions and 3 deletions in the isolate compared with GenBank accession number L22866. The result indicated some functional disorder in the gene. Although the CACA plate is an excellent selective medium for Cr. neoformans, other identification methods should also be used. PMID- 11479534 TI - [Observation of tinea unguium by neutral red staining]. AB - To evaluate the viability of dermatophytes in tinea unguium, we applied neutral red staining (NRS) for nail powder obtained from direct microscopic examination (DME) of positive tinea unguium patients. The nail powder from tinea unguium patients by grinder was applied following three sets of examinations, NRS, DME, and culture on Mycosel medium. The positive rates of nail powder obtained from 50 tinea unguium patients were 35 (70%), 37 (74%) and 2 (4%), respectively. Correlation of three sets of examinations were as follows: twenty-five patients were NRS positive, DME positive and culture negative, 10 patients were NRS negative, DME positive and culture negative, and 8 patients were NRS positive, DME negative and culture negative. These findings suggest that application of NRS for nail powder from tinea unguium patients is a practical method for evaluation of the viability of fungal elements. PMID- 11479535 TI - Nocardia brasiliensis infection seen on grafted skin of the dorsum of a foot. AB - For the past 4 years a 23 y-old female has noticed erythema on the dorsum of the right foot, where skin was grafted due to a traffic accident 20 years ago. She visited the Department of Dermatology of Gifu Prefectural Hospital on Oct. 19, 1998; her general health was good. Physical examination disclosed a swelling with erythema, papules and pustules on the dorsum of the right foot. The results of routine laboratory investigations were within normal limits except for the white blood cell count (11,300/mm(3)), blood sediment rate (25 mm/hrs), C reactive protein (1.21) and rheumatoid factor (x 16.6). Several yellowish and verrucous or wrinkled colonies were grown on Sabourauds agar culture from the biopsied specimen of the foot. Histopathological features showed epidermal hyperplasia with elongation of rete ridges and granulomatous changes in the dermis; many mononuclear and giant cells were present, and several positive fine filamentous and irregularly branching structures with PAS and Grocott stains were seen in the granulomatous nests. Both clinical and histopathological features led to speculation of Nocardia infection, and Nocardia brasiliensis was determined. The patient was treated by surgical total resection including the grafted skin. Although a soybean-sized nodule was seen on the border of the skin-graft of the foot three months later, there was no recurrence after the local resection. PMID- 11479536 TI - A human case of protothecosis successfully treated with itraconazole. AB - Cutaneous protothecosis developed in a 63-year-old Japanese female. The patient had a long history of steroid use for bronchial asthma. A tender, swollen, erythematous plaque with white papules covered the dorsal aspect of the patients right hand. Histopathological examination showed typical protothecosis with numerous mulberry-like sporangia positive for periodic acid-Schiff stain. The pathogen was identified as Prototheca wickerhamii. After a 6-week course of 150 200 mg/day itraconazole (ITCZ) therapy, the patients culture was negative for P. wickerhamii. No recurrence was seen after 12 weeks of ITCZ therapy. This case of protothecosis and those of 16 patients are summarized. All the patients had lived in mid- to southern Japan. Only 12 cases of protothecosis treated with ITCZ have been reported in the literature; 8 of these cases were cured with ITCZ therapy alone. In cases treated with ITCZ at no more than 100 mg/day, medication was typically needed for 2 months; most of the cases treated with ITCZ at 200 mg/day were successfully cured within 2 months. A 2-month course of ITCZ at 200 mg/day appears to be adequate treatment for human protothecosis. PMID- 11479537 TI - [A case of lymphocutaneous sporotrichosis]. AB - We report a case of lymphocutaneous sporotrichosis in a 69-year old man who had nodular-ulcerated lesions on the right hand and forearm. Small nodules remained on the right hand after 8 weeks of 0.5 g daily treatment with potassium iodide. Alternatively, terbinafine therapy (125 mg/day) resulted in healing with atrophic scars after 9 weeks without side effects. We reviewed 67 patients of cutaneous sporotrichosis in Japan from 1993 to 1999. Those cure rates (and mean durations of treatments in parentheses) are 90.9% (8.1 weeks) with potassium iodide, 86.6% (12.6 weeks) with itraconazole (100 mg/day) and 71.4% (12.8 weeks) with terbinafine (125 mg/day) treatments, respectively. These results lead us to consider a daily dose of 250 mg as more appropriate for terbinafine. PMID- 11479540 TI - Rights of access. PMID- 11479539 TI - Identification of the gene altered in Berardinelli-Seip congenital lipodystrophy on chromosome 11q13. AB - Congenital generalized lipodystrophy, or Berardinelli-Seip syndrome (BSCL), is a rare autosomal recessive disease characterized by a near-absence of adipose tissue from birth or early infancy and severe insulin resistance. Other clinical and biological features include acanthosis nigricans, hyperandrogenism, muscular hypertrophy, hepatomegaly, altered glucose tolerance or diabetes mellitus, and hypertriglyceridemia. A locus (BSCL1) has been mapped to 9q34 with evidence of heterogeneity. Here, we report a genome screen of nine BSCL families from two geographical clusters (in Lebanon and Norway). We identified a new disease locus, designated BSCL2, within the 2.5-Mb interval flanked by markers D11S4076 and D11S480 on chromosome 11q13. Analysis of 20 additional families of various ethnic origins led to the identification of 11 families in which the disease cosegregates with the 11q13 locus; the remaining families provide confirmation of linkage to 9q34. Sequence analysis of genes located in the 11q13 interval disclosed mutations in a gene homologous to the murine guanine nucleotide-binding protein (G protein), gamma3-linked gene (Gng3lg) in all BSCL2-linked families. BSCL2 is most highly expressed in brain and testis and encodes a protein (which we have called seipin) of unknown function. Most of the variants are null mutations and probably result in a severe disruption of the protein. These findings are of general importance for understanding the molecular mechanisms underlying regulation of body fat distribution and insulin resistance. PMID- 11479541 TI - Monstrous crowing. PMID- 11479542 TI - Fickle financing climate continues. PMID- 11479543 TI - Biogen, Serono squabble over MS market. PMID- 11479544 TI - US, Brazil tangle over IP rights to drugs. PMID- 11479545 TI - Industry-academia study lacking, say critics. PMID- 11479546 TI - FoE urges UK to disclose more of GM crop trial data. PMID- 11479547 TI - Jury out on environmental impact of GM soy. PMID- 11479548 TI - Celera buys Axys. PMID- 11479549 TI - Redefining priorities in gene-based drug discovery. PMID- 11479550 TI - An industry imperiled by regulatory bottlenecks. PMID- 11479552 TI - Is VP22 nuclear homing an artifact? PMID- 11479554 TI - FAO/WHO protocol abandons science. PMID- 11479555 TI - The production capacity bottleneck. PMID- 11479556 TI - Quality control of plasmid preparations. PMID- 11479557 TI - Microfluidics--downsizing large-scale biology. PMID- 11479558 TI - ADMET--turning chemicals into drugs. PMID- 11479559 TI - Drug discovery--an operating model for a new era. PMID- 11479560 TI - Interrogation by toxin. PMID- 11479561 TI - A PURE approach to constructive biology. PMID- 11479562 TI - Phage antibacterials make a comeback. PMID- 11479563 TI - Silencing the majority. PMID- 11479564 TI - Antisense that comes naturally. PMID- 11479565 TI - Structures without crystals. PMID- 11479566 TI - Pigs expressing salivary phytase produce low-phosphorus manure. AB - To address the problem of manure-based environmental pollution in the pork industry, we have developed the phytase transgenic pig. The saliva of these pigs contains the enzyme phytase, which allows the pigs to digest the phosphorus in phytate, the most abundant source of phosphorus in the pig diet. Without this enzyme, phytate phosphorus passes undigested into manure to become the single most important manure pollutant of pork production. We show here that salivary phytase provides essentially complete digestion of dietary phytate phosphorus, relieves the requirement for inorganic phosphate supplements, and reduces fecal phosphorus output by up to 75%. These pigs offer a unique biological approach to the management of phosphorus nutrition and environmental pollution in the pork industry. PMID- 11479567 TI - Diphtheria toxin receptor-mediated conditional and targeted cell ablation in transgenic mice. AB - Specific cell ablation is a useful method for analyzing the in vivo function of cells. We have developed a simple and sensitive method for conditional cell ablation in transgenic mice, called "toxin receptor-mediated cell knockout." We expressed the diphtheria toxin (DT) receptor in transgenic mice using a hepatocyte-specific promoter and found that injection of DT caused fulminant hepatitis. Three independently established transgenic lines demonstrated a good correlation between the sensitivity of hepatocytes to DT and the expression level of the DT receptors. Moreover, the degree of hepatocyte damage was easily controlled over a wide range of doses of injected DT without any obvious abnormalities in other cells or tissues. This system is useful for generating mouse models of disease and for studying the recovery or regeneration of tissues from cell damage or loss. As DT is a potent inhibitor of protein synthesis in both growing and non-growing cells, the method is applicable to a wide range of cells and tissues in mice or in other DT-insensitive animals. PMID- 11479568 TI - Cell-free translation reconstituted with purified components. AB - We have developed a protein-synthesizing system reconstituted from recombinant tagged protein factors purified to homogeneity. The system was able to produce protein at a rate of about 160 microg/ml/h in a batch mode without the need for any supplementary apparatus. The protein products were easily purified within 1 h using affinity chromatography to remove the tagged protein factors. Moreover, omission of a release factor allowed efficient incorporation of an unnatural amino acid using suppressor transfer RNA (tRNA). PMID- 11479569 TI - Combinatorial fluorescence energy transfer tags for multiplex biological assays. AB - We report an approach for developing combinatorial fluorescence energy transfer (CFET) tags by tuning the tags' fluorescence emission signatures. The tags can all be excited at a single wavelength and analyzed by a simple optical system. We constructed eight CFET tags with unique fluorescence signatures, detected by a three-color capillary array electrophoresis (CAE) system with 488 nm excitation, using only three fluorescent dyes. A 1',2'-dideoxyribose phosphate spacer was used to separate the donor and acceptor to tune the energy transfer efficiency, generating unique fluorescence signatures. The spacer also served as an electrophoretic mobility tag to tune the mobility of CFET-labeled DNA for multiplex detection of single-nucleotide polymorphisms (SNPs). Six nucleotide variations were identified simultaneously using six CFET tags on synthetic DNA templates and on a PCR product from the retinoblastoma tumor suppressor gene. PMID- 11479570 TI - Plant viral genes in DNA idiotypic vaccines activate linked CD4+ T-cell mediated immunity against B-cell malignancies. AB - DNA delivery of tumor antigens can activate specific immune attack on cancer cells. However, antigens may be weak, and immune capacity can be compromised. Fusion of genes encoding activating sequences to the tumor antigen sequence facilitates promotion and manipulation of effector pathways. Idiotypic determinants of B-cell tumors, encoded by the variable region genes, are clone specific tumor antigens. When assembled as single-chain Fv (scFv) alone in a DNA vaccine, immunogenicity is low. Previously, we found that fusion of a sequence from tetanus toxin (fragment C; FrC) promoted anti-idiotypic protection against lymphoma and myeloma. We have now investigated an alternative fusion gene derived from a plant virus, potato virus X coat protein, a primary antigen in humans. When fused to scFv, the self-aggregating protein generates protection against lymphoma and myeloma. In contrast to scFv-FrC, protection against lymphoma is mediated by CD4+ T cells, as is protection against myeloma. Plant viral proteins offer new opportunities to activate immunity against linked T-cell epitopes to attack cancer. PMID- 11479571 TI - Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit. AB - Transgenic tomato plants overexpressing a vacuolar Na+/H+ antiport were able to grow, flower, and produce fruit in the presence of 200 mM sodium chloride. Although the leaves accumulated high sodium concentrations, the tomato fruit displayed very low sodium content. Contrary to the notion that multiple traits introduced by breeding into crop plants are needed to obtain salt-tolerant plants, the modification of a single trait significantly improved the salinity tolerance of this crop plant. These results demonstrate that with a combination of breeding and transgenic plants it could be possible to produce salt-tolerant crops with far fewer target traits than had been anticipated. The accumulation of sodium in the leaves and not in the fruit demonstrates the utility of such a modification in preserving the quality of the fruit. PMID- 11479572 TI - Direct visualization of protein interactions in plant cells. AB - The protein NPR1/NIM1 is required for the induction of systemic acquired resistance (SAR) in plants and has been shown to interact with members of the TGA/OBF family of basic leucine zipper (bZIP) transcription factors. However, to date, there is no method available to monitor such interactions in plant cells. We report here an in vivo protein fragment complementation assay (PCA), based on association of reconstituted murine dihydrofolate reductase (mDHFR) with a fluorescent probe to detect protein-protein interaction in planta. We demonstrate that the interaction between Arabidopsis NPR1/NIM1 and the bZIP factor TGA2 is induced by the regulators of SAR, salicylic acid (SA), and its analog 2,6 dichloroisonicotinic acid (INA) with distinct species-specific responses. Furthermore, the induced interaction is localized predominantly in the nucleus. Protein fragment complementation assays could be of value to agricultural research by providing a system for high-throughput biochemical pathway mapping and for screening of small molecules that modulate protein interactions. PMID- 11479573 TI - In vivo site-directed mutagenesis using oligonucleotides. AB - Functional characterization of the genes of higher eukaryotes has been aided by their expression in model organisms and by analyzing site-specific changes in homologous genes in model systems such as the yeast Saccharomyces cerevisiae. Modifying sequences in yeast or other organisms such that no heterologous material is retained requires in vitro mutagenesis together with subcloning. PCR based procedures that do not involve cloning are inefficient or require multistep reactions that increase the risk of additional mutations. An alternative approach, demonstrated in yeast, relies on transformation with an oligonucleotide, but the method is restricted to the generation of mutants with a selectable phenotype. Oligonucleotides, when combined with gap repair, have also been used to modify plasmids in yeast; however, this approach is limited by restriction-site availability. We have developed a mutagenesis approach in yeast based on transformation by unpurified oligonucleotides that allows the rapid creation of site-specific DNA mutations in vivo. A two-step, cloning-free process, referred to as delitto perfetto, generates products having only the desired mutation, such as a single or multiple base change, an insertion, a small or a large deletion, or even random mutations. The system provides for multiple rounds of mutation in a window up to 200 base pairs. The process is RAD52 dependent, is not constrained by the distribution of naturally occurring restriction sites, and requires minimal DNA sequencing. Because yeast is commonly used for random and selective cloning of genomic DNA from higher eukaryotes such as yeast artificial chromosomes, the delitto perfetto strategy also provides an efficient way to create precise changes in mammalian or other DNA sequences. PMID- 11479574 TI - Patenting expressed sequence tags and single nucleotide polymorphisms. PMID- 11479575 TI - Recent patents in automation in drug discovery. PMID- 11479578 TI - How to mismanage a search from start to finish. PMID- 11479579 TI - Defining a new bioethic. PMID- 11479580 TI - Iron on the brain. AB - Accumulations of iron are often detected in the brains of people suffering from neurodegenerative diseases. But it is often not known whether such accumulations contribute directly to disease progression. The identification of the genes mutated in two such disorders suggests that errors in iron metabolism do indeed have a key role. PMID- 11479581 TI - Eggs in the balance. AB - Polycyclic aromatic hydrocarbons (PAH), found in cigarette smoke and air pollution, interact with the aryl hydrocarbon receptor (Ahr) to cause reproductive defects. Mice lacking either Ahr or the pro-apoptotic protein Bax have an increased number of primordial follicles, and these mutant oocytes are resistant to PAH toxicity. A new study shows that the Bax promoter contains two core Ahr response elements, which are required for PAH stimulation of Bax promoter activity in oocytes. Thus, the toxic effects of PAH in oocytes are mediated directly by Ahr induction of the Bax pathway. PMID- 11479582 TI - Medaka on the move. PMID- 11479583 TI - To bind or not to bind. AB - Gene expression is regulated by transcription factors binding selectively to particular portions of the genome. To what extent are these protein-DNA interactions influenced by the intrinsic sequence-specific recognition properties at each protein, and to what extent are they affected by other factors, such as chromatin structure or cooperative interactions with other proteins. Genome-wide surveys of DNA binding by transcription factors in vivo are beginning to provide some answers. PMID- 11479587 TI - The value of isolated populations. PMID- 11479588 TI - DelBank: a mouse ES-cell resource for generating deletions. PMID- 11479589 TI - Regulating use of stem cells. PMID- 11479590 TI - Mutation of DNASE1 in people with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a highly prevalent human autoimmune diseases that causes progressive glomerulonephritis, arthritis and an erythematoid rash. Mice deficient in deoxyribonuclease I (Dnase1) develop an SLE like syndrome. Here we describe two patients with a heterozygous nonsense mutation in exon 2 of DNASE1, decreased DNASE1 activity and an extremely high immunoglobulin G titer against nucleosomal antigens. These data are consistent with the hypothesis that a direct connection exists between low activity of DNASE1 and progression of human SLE. PMID- 11479591 TI - IBD sharing around the PPARG locus is not increased in dizygotic twins or their mothers. AB - Busjahn et al reported finding linkage between PPARG (3p25) and being a dizygotic (DZ) twin. We differentiate, as do the authors, between the conception of DZ twins, and being the viable result of such a conception. PMID- 11479592 TI - Photo-mediated gene activation using caged RNA/DNA in zebrafish embryos. AB - We report a new and simple technique for photo-mediated temporal and spatial control of gene activation in zebrafish embryos as an alternative to the gene 'knockdown' approach using antisense, morpholino-modified oligonucleotides (morpholinos). The synthetic compound 6-bromo-4-diazomethyl-7-hydroxycoumarin (Bhc-diazo) forms a covalent bond with the phosphate moiety of the sugar phosphate backbone of RNA, a process known as caging. The 6-bromo-7 hydroxycoumarin-4-ylmethyl (Bhc) group binds to approximately 30 sites on the phosphate moieties per 1 kb of RNA sequence. Bhc-caged mRNA undergoes photolysis (uncaging) when exposed to long-wave ultraviolet light (350 to 365 nm). We show that Bhc-caged green fluorescent protein (Gfp) mRNA has severely reduced translational activity in vitro, whereas illumination of Bhc-caged mRNA with ultraviolet light leads to partial recovery of translational activity. Bhc-caged mRNA is highly stable in zebrafish embryos. In embryos injected with Bhc-caged Gfp mRNA at the one-cell stage, GFP protein expression and fluorescence is specifically induced by ultraviolet light. We also show that, consistent with results obtained using other methods, uncaging eng2a (which encodes the transcription factor Engrailed2a) in the head region during early development causes a severe reduction in the size of the eye and enhanced development of the midbrain and the midbrain-hindbrain boundary at the expense of the forebrain. PMID- 11479593 TI - CTCF-binding sites flank CTG/CAG repeats and form a methylation-sensitive insulator at the DM1 locus. AB - An expansion of a CTG repeat at the DM1 locus causes myotonic dystrophy (DM) by altering the expression of the two adjacent genes, DMPK and SIX5, and through a toxic effect of the repeat-containing RNA. Here we identify two CTCF-binding sites that flank the CTG repeat and form an insulator element between DMPK and SIX5. Methylation of these sites prevents binding of CTCF, indicating that the DM1 locus methylation in congenital DM would disrupt insulator function. Furthermore, CTCF-binding sites are associated with CTG/CAG repeats at several other loci. We suggest a general role for CTG/CAG repeats as components of insulator elements at multiple sites in the human genome. PMID- 11479594 TI - A novel pantothenate kinase gene (PANK2) is defective in Hallervorden-Spatz syndrome. AB - Hallervorden-Spatz syndrome (HSS) is an autosomal recessive neurodegenerative disorder associated with iron accumulation in the brain. Clinical features include extrapyramidal dysfunction, onset in childhood, and a relentlessly progressive course. Histologic study reveals iron deposits in the basal ganglia. In this respect, HSS may serve as a model for complex neurodegenerative diseases, such as Parkinson disease, Alzheimer disease, Huntington disease and human immunodeficiency virus (HIV) encephalopathy, in which pathologic accumulation of iron in the brain is also observed. Thus, understanding the biochemical defect in HSS may provide key insights into the regulation of iron metabolism and its perturbation in this and other neurodegenerative diseases. Here we show that HSS is caused by a defect in a novel pantothenate kinase gene and propose a mechanism for oxidative stress in the pathophysiology of the disease. PMID- 11479595 TI - Imprinted X inactivation maintained by a mouse Polycomb group gene. AB - In mammals, dosage compensation of X-linked genes is achieved by the transcriptional silencing of one X chromosome in the female (reviewed in ref. 1). This process, called X inactivation, is usually random in the embryo proper. In marsupials and the extra-embryonic region of the mouse, however, X inactivation is imprinted: the paternal X chromosome is preferentially inactivated whereas the maternal X is always active. Having more than one active X chromosome is deleterious to extra-embryonic development in the mouse. Here we show that the gene eed (embryonic ectoderm development), a member of the mouse Polycomb group (Pc-G) of genes, is required for primary and secondary trophoblast giant cell development in female embryos. Results from mice carrying a paternally inherited X-linked green fluorescent protein (GFP) transgene implicate eed in the stable maintenance of imprinted X inactivation in extra-embryonic tissues. Based on the recent finding that the Eed protein interacts with histone deacetylases, we suggest that this maintenance activity involves hypoacetylation of the inactivated paternal X chromosome in the extra-embryonic tissues. PMID- 11479596 TI - Mutations in the gene encoding B, a novel transporter protein, reduce melanin content in medaka. AB - Pigmentation of the skin is of great social, clinical and cosmetic significance. Several genes that, when mutated, give rise to altered coat color in mice have been identified; their analysis has provided some insight into melanogenesis and human pigmentation diseases. Such analyses do not, however, fully inform on the pigmentation of lower vertebrates because mammals have only one kind of chromatophore, the melanocyte. In contrast, the medaka (a small, freshwater teleost) is a suitable model of the lower vertebrates because it has all kinds of chromatophores. The basic molecular genetics of fish are known and approximately 70 spontaneous pigmentation mutants have been isolated. One of these, an orange red variant, is a homozygote of a well-known and common allele, b, and has been bred for hundreds of years by the Japanese. Here, we report the first successful positional cloning of a medaka gene (AIM1): one that encodes a transporter that mediates melanin synthesis. The protein is predicted to consist of 12 transmembrane domains and is 55% identical to a human EST of unknown function isolated from melanocytes and melanoma cells. We also isolated a highly homologous gene from the mouse, indicating a conserved function of vertebrate melanogenesis. Intriguingly, these proteins have sequence and structural similarities to plant sucrose transporters, suggesting a relevance of sucrose in melanin synthesis. Analysis of AIM1 orthologs should provide new insights into the regulation of melanogenesis in both teleosts and mammals. PMID- 11479598 TI - In science we trust. PMID- 11479597 TI - Mutations in the region encoding the von Willebrand factor A domain of matrilin-3 are associated with multiple epiphyseal dysplasia. AB - Multiple epiphyseal dysplasia (MED) is a relatively mild and clinically variable osteochondrodysplasia, primarily characterized by delayed and irregular ossification of the epiphyses and early-onset osteoarthritis. Mutations in the genes encoding cartilage oligomeric matrix protein (COMP) and type IX collagen (COL9A2 and COL9A3) have previously been shown to cause different forms of MED (refs. 4-13). These dominant forms of MED (EDM1-3) are caused by mutations in the genes encoding structural proteins of the cartilage extracellular matrix (ECM); these proteins interact with high affinity in vitro. A recessive form of MED (EDM4) has also been reported; it is caused by a mutation in the diastrophic dysplasia sulfate transporter gene (SLC26A). A genomewide screen of family with autosomal-dominant MED not linked to the EDM1-3 genes provides significant genetic evidence for a MED locus on the short arm of chromosome 2 (2p24-p23), and a search for candidate genes identified MATN3 (ref. 18), encoding matrilin-3, within the critical region. Matrilin-3 is an oligomeric protein that is present in the cartilage ECM. We have identified two different missense mutations in the exon encoding the von Willebrand factor A (vWFA) domain of matrilin-3 in two unrelated families with MED (EDM5). These are the first mutations to be identified in any of the genes encoding the matrilin family of proteins and confirm a role for matrilin-3 in the development and homeostasis of cartilage and bone. PMID- 11479599 TI - Optimizing human fertility and survival. PMID- 11479600 TI - Misconduct trouble brewing in Gottingen. PMID- 11479601 TI - Stem cell researchers ponder worst-case scenario. PMID- 11479604 TI - Report predicts burgeoning vaccine markets. PMID- 11479605 TI - Trials suspended due to death at Hopkins. PMID- 11479607 TI - Lassa fever is unheralded problem in West Africa. PMID- 11479608 TI - Update on American death, disease and exercise. PMID- 11479609 TI - Alfred Gilman. AB - At the age of 10, Alfred Gilman wanted to go to the moon; on a visit to New York's Hayden Planetarium with his parents, he signed up to be an astronaut. Fifty years later, Gilman is preparing to take on a different, yet equally astronomical task. Instead of traveling to outer space, he is heading up a research program to detail the workings of cellular space. PMID- 11479610 TI - Less is more? STI in acute and chronic HIV-1 infection. PMID- 11479613 TI - You are what you secrete. PMID- 11479614 TI - Cystic fibrosis salt/fluid controversy: in the thick of it. PMID- 11479615 TI - A mouse model for hepatitis C virus infection? PMID- 11479616 TI - Anticancer squirt. PMID- 11479617 TI - The economy of T-cell memory: CD4+ recession in times of CD8+ stability? PMID- 11479618 TI - TB comes to a sticky beginning. PMID- 11479619 TI - Might mammalian mitochondria merge? PMID- 11479621 TI - Autoimmune disease: why and where it occurs. AB - Autoimmune disease is controlled by genetic and environmental factors. Both of these affect susceptibility to autoimmunity at three levels: the overall reactivity of the immune system, the specific antigen and its presentation, and the target issue. PMID- 11479622 TI - Signaling through OX40 (CD134) breaks peripheral T-cell tolerance. AB - Peripheral T-cell tolerance is a mechanism to limit autoimmunity, but represents a major obstacle in diseases such as cancer. Tolerance is due to limited accumulation of antigen-specific T cells accompanied by functional hypo responsiveness, and is induced by antigen encounter in a non-inflammatory environment. In contrast to advances in preventing induction of T-cell tolerance, there has been little progress in defining targets to reverse established tolerance. Here we show that signals from a single dose of an agonistic antibody against OX40 (CD134, a member of the tumor necrosis-factor family of receptors) can break an existing state of tolerance in the CD4+ T-cell compartment. OX40 signals promote T-cell expansion after the hypo-responsive phenotype is induced and restore normal functionality. These data highlight the potent costimulatory capacity of OX40, and indicate OX40 as a target for therapeutic intervention in a variety of related diseases. PMID- 11479623 TI - Differential regulation of antiviral T-cell immunity results in stable CD8+ but declining CD4+ T-cell memory. AB - Emerging evidence indicates that CD8+ and CD4+ T-cell immunity is differentially regulated. Here we have delineated differences and commonalities among antiviral T-cell responses by enumeration and functional profiling of eight specific CD8+ and CD4+ T-cell populations during primary, memory and recall responses. A high degree of coordinate regulation among all specific T-cell populations stood out against an approximately 20-fold lower peak expansion and prolonged contraction phase of specific CD4+ T-cell populations. Surprisingly, although CD8+ T-cell memory was stably maintained for life, levels of specific CD4+ memory T cells gradually declined. However, this decay, which seemed to result from less efficient rescue from apoptosis, did not affect functionality of surviving virus specific CD4+ T cells. Our results indicate that CD4+ T-cell memory might become limiting under physiological conditions and that conditions precipitating CD4+ T cell loss might compromise protective immunity even in the presence of unimpaired CD8+ T-cell responses. PMID- 11479624 TI - Inhibition of HIV-1 virion production by a transdominant mutant of integrase interactor 1. AB - Integase interactor 1 (INI1), also known as hSNF5, is a protein that interacts with HIV-1 integrase. We report here that a cytoplasmically localized fragment of INI1 (S6; aa183-294) containing the minimal integrase-interaction domain potently inhibits HIV-1 particle production and replication. Mutations in S6 or integrase that disrupt integrase-INI1 interaction abrogated the inhibitory effect. An integrase-deficient HIV-1 transcomplemented with integrase fused to Vpr was not affected by S6. INI1 was specifically incorporated into virions and was required for efficient HIV-1 particle production. These results indicate that INI1 is required for late events in the viral life cycle, and that ectopic expression of S6 inhibits HIV-1 replication in a transdominant manner via its specific interaction with integrase within the context of Gag-Pol, providing a novel strategy to control HIV-1 replication. PMID- 11479625 TI - Hepatitis C virus replication in mice with chimeric human livers. AB - Lack of a small animal model of the human hepatitis C virus (HCV) has impeded development of antiviral therapies against this epidemic infection. By transplanting normal human hepatocytes into SCID mice carrying a plasminogen activator transgene (Alb-uPA), we generated mice with chimeric human livers. Homozygosity of Alb-uPA was associated with significantly higher levels of human hepatocyte engraftment, and these mice developed prolonged HCV infections with high viral titers after inoculation with infected human serum. Initial increases in total viral load were up to 1950-fold, with replication confirmed by detection of negative-strand viral RNA in transplanted livers. HCV viral proteins were localized to human hepatocyte nodules, and infection was serially passaged through three generations of mice confirming both synthesis and release of infectious viral particles. These chimeric mice represent the first murine model suitable for studying the human hepatitis C virus in vivo. PMID- 11479626 TI - Inter-mitochondrial complementation: Mitochondria-specific system preventing mice from expression of disease phenotypes by mutant mtDNA. AB - Here we investigated the pathogenesis of deletion mutant mitochondrial (mt)DNA by generating mice with mutant mtDNA carrying a 4696-basepair deletion (DeltamtDNA4696), and by using cytochrome c oxidase (COX) electron micrographs to identify COX activity at the individual mitochondrial level. All mitochondria in tissues with DeltamtDNA4696 showed normal COX activity until DeltamtDNA4696 accumulated predominantly; this prevented mice from expressing disease phenotypes. Moreover, we did not observe coexistence of COX-positive and negative mitochondria within single cells. These results indicate the occurrence of inter-mitochondrial complementation through exchange of genetic contents between exogenously introduced mitochondria with DeltamtDNA4696 and host mitochondria with normal mtDNA. This complementation shows a mitochondria specific mechanism for avoiding expression of deletion-mutant mtDNA, and opens the possibility of a gene therapy in which mitochondria possessing full-length DNA are introduced. PMID- 11479627 TI - The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity. AB - Adiponectin is an adipocyte-derived hormone. Recent genome-wide scans have mapped a susceptibility locus for type 2 diabetes and metabolic syndrome to chromosome 3q27, where the gene encoding adiponectin is located. Here we show that decreased expression of adiponectin correlates with insulin resistance in mouse models of altered insulin sensitivity. Adiponectin decreases insulin resistance by decreasing triglyceride content in muscle and liver in obese mice. This effect results from increased expression of molecules involved in both fatty-acid combustion and energy dissipation in muscle. Moreover, insulin resistance in lipoatrophic mice was completely reversed by the combination of physiological doses of adiponectin and leptin, but only partially by either adiponectin or leptin alone. We conclude that decreased adiponectin is implicated in the development of insulin resistance in mouse models of both obesity and lipoatrophy. These data also indicate that the replenishment of adiponectin might provide a novel treatment modality for insulin resistance and type 2 diabetes. PMID- 11479628 TI - The adipocyte-secreted protein Acrp30 enhances hepatic insulin action. AB - Acrp30 is a circulating protein synthesized in adipose tissue. A single injection in mice of purified recombinant Acrp30 leads to a 2-3-fold elevation in circulating Acrp30 levels, which triggers a transient decrease in basal glucose levels. Similar treatment in ob/ob, NOD (non-obese diabetic) or streptozotocin treated mice transiently abolishes hyperglycemia. This effect on glucose is not associated with an increase in insulin levels. Moreover, in isolated hepatocytes, Acrp30 increases the ability of sub-physiological levels of insulin to suppress glucose production. We thus propose that Acrp30 is a potent insulin enhancer linking adipose tissue and whole-body glucose metabolism. PMID- 11479629 TI - Tumoricidal activity of a novel anti-human DR5 monoclonal antibody without hepatocyte cytotoxicity. AB - A novel anti-human DR5 monoclonal antibody, TRA-8, induces apoptosis of most tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-sensitive tumor cells both in vitro and in vivo. In contrast to both the membrane-bound form of human TRAIL, which induced severe hepatitis in mice, and the soluble form of human TRAIL, which induced apoptosis of normal human hepatocytes in vitro, TRA-8 did not induce significant cell death of normal human hepatocytes. However, both primary hepatocellular carcinoma cells and an established liver cancer cell line were highly susceptible to the killing mediated by TRA-8. We show here that elevated levels of cell-surface expression of DR5 and increased susceptibility to DR5-mediated apoptosis are characteristics of malignant tumor cells. In contrast, DR5 alone is not sufficient to trigger apoptosis of normal hepatocytes. Therefore, selective, specific targeting of DR5 with an agonistic antibody might be a safe and effective strategy for cancer therapy. PMID- 11479630 TI - Antiproliferative activity of ecteinascidin 743 is dependent upon transcription coupled nucleotide-excision repair. AB - While investigating the novel anticancer drug ecteinascidin 743 (Et743), a natural marine product isolated from the Caribbean sea squirt, we discovered a new cell-killing mechanism mediated by DNA nucleotide excision repair (NER). A cancer cell line selected for resistance to Et743 had chromosome alterations in a region that included the gene implicated in the hereditary disease xeroderma pigmentosum (XPG, also known as Ercc5). Complementation with wild-type XPG restored the drug sensitivity. Xeroderma pigmentosum cells deficient in the NER genes XPG, XPA, XPD or XPF were resistant to Et743, and sensitivity was restored by complementation with wild-type genes. Moreover, studies of cells deficient in XPC or in the genes implicated in Cockayne syndrome (CSA and CSB) indicated that the drug sensitivity is specifically dependent on the transcription-coupled pathway of NER. We found that Et743 interacts with the transcription-coupled NER machinery to induce lethal DNA strand breaks. PMID- 11479631 TI - Lethality-based selection of recombinant genes in mammalian cells: application to identifying tumor antigens. AB - Many biological processes result in either cell death or cessation of cell growth. However, plasmid- and retrovirus-based mammalian expression vectors in which it has been possible to construct representative cDNA libraries cannot be readily recovered from cells that are not actively dividing. This has limited the efficiency of selection of recombinant genes that mediate either lytic events or growth arrest. Examples include genes that encode the target antigens of cytotoxic T cells, genes that promote stem-cell differentiation and pro-apoptotic genes. We have successfully constructed representative cDNA libraries in a poxvirus-based vector that can be recovered from cells that have undergone lethality-based selection. This strategy has been applied to selection of a gene that encodes a cytotoxic T-cell target antigen common to several independently derived tumors. PMID- 11479634 TI - Necrotizing fasciitis. AB - Necrotizing fasciitis is a devestating condition which has been recognized for several years. In North America a recent increase of cases has led to much media attention and public fear. Necrotizing fasciitis may occur as a consequence of infection with Streptococcus pyogenes or as a result of a polymicrobial synergistic infection caused by aerobic, anaerobic, gram positive and gram negative organisms, often in postoperative patients. Necrotizing fasciitis caused by Streptococcus pyogenes is mediated by superantigens. The management of necrotizing fasciitis requires a high index of suspicion for diagnosis followed by antimicrobial therapy and early surgical intervention. In cases caused by Streptococcus pyogenes with streptococcal toxic shock syndrome, intravenous immunoglobulin may be of benefit. PMID- 11479635 TI - Antimicrobial susceptibility testing and patterns of resistance at a tertiary care center. AB - Clinical microbiology laboratories are faced with the challenge of accurately detecting emerging antibiotic resistance in bacterial pathogens. In recent years, vancomycin resistant enterococci have emerged, as have penicillin resistant pneumococci and more recently, methicillin-resistant Staphylococcus aureus with reduced susceptibility to vancomycin. In order to detect these emerging resistant pathogens it is essential that antimicrobial susceptibility be carried out by laboratories as an integral part of therapeutic strategies. In this review, we discuss patterns of susceptibility of different antimicrobials as experienced at King Faisal Specialist Hospital and Research Centre, a tertiary care center in Riyadh. PMID- 11479636 TI - Brucellosis among animals and human contacts in eastern Sudan. AB - OBJECTIVE: To study the relative frequency of brucellosis among domestic animals in Kassala State, Sudan, in the year 1999 and compare the results of previous studies since 1908. Also to study the frequency rates of the disease in animal contacts in the area. METHODS: Sero-survey for Brucella melitensis and Brucella abortus was carried out on sera of animals brought for slaughter to Kassala abattoir and on sera of occupational contacts of animals. All sera were tested by the slide agglutination test. The positive reactors were confirmed by tube agglutination test. RESULTS: A total of 1225 sera were tested - 1038 were animal sera and 187 were human sera. Four percent of goats sera, 1% of sheep sera and 5% of cattle sera were found to be positive. Of the 64 camel sera tested, none were positive reactors. Of the occupational contacts, which included butchers, slaughterhouse workers, milkers and cow attendants, (1%) reacted positively. CONCLUSION: The study showed low frequency rates of brucellosis among animals in the Kassala area compared with other parts of Sudan. Occupational contacts showed very low frequency rates of the disease. This is often over-looked by medical practitioners due to the overwhelming problem of malaria in the area. We draw the attention of practitioners to think of brucellosis in the differential diagnosis of fever especially in rural communities. PMID- 11479637 TI - Teenagers obstetric performance. AB - OBJECTIVE: To study the pregnancy outcome in teenage primigravida women admitted and delivered in our unit between April 1997 and March 1998, and to compare the outcome with other primigravida above 20 years old. METHODS: Data was collected from primigravidae in respect of age, last menstrual period, history of booking at Primary Health Center and complications of pregnancies were identified. The process of labor and delivery were monitored and the outcome recorded. RESULTS: During the period of study, 2,650 women delivered in the unit, 171 (6%) were primigravida, out of which 116 (68%) were teenagers, ages between 13 and 19 years old, 55 (32%) were above 20 years of age. All the primigravidas were married and therefore had their husband and parental support. Forty three percent of the teenagers plan to return to school after delivery. The length of the 2nd stage of labor (67.7 minutes) in the young teenagers aged 13 to 15 years was significantly longer than of the older teenagers 16 to 19 years old and that of the control group ages above 20 years old, P<0.0001. The mean birth weight (2.45 kgm) in the younger teenagers were also lower than that of the older teenagers and the control group (3 kgm and 3.25 kgm) P<0.0001. There was no significant difference between the teenagers ages 13 to 19 years old and the control group regarding normal vagina delivery, lower segment cesarean, ventouse delivery, number of anemic patients and the mean birth weight as shown by the P-values. There was no significant difference in the numbers and types of medical complications identified between the teenagers and the control group. CONCLUSION: The younger teenage group (13 to 15 years) has been identfied as the high-risk group in this study but there was no significant difference in the pregnancy outcome of the teenagers (13 to 19 years old) in general compared with the control group. Attention must therefore be turned to the young teenagers pregnancy, labor and delivery. To avoid poor outcome in this age group, age at first pregnancy should be encouraged from 16 years and above. PMID- 11479638 TI - Ultrasound in the management of Chorioangioma. AB - OBJECTIVE: To evaluate the role of ultrasound in the diagnosis and prediction of the clinical course of a large chorioangioma. METHODS: Six women with a large chorioangioma were consecutively detected at 26 28 weeks gestation by ultrasound. At initial examination, the tumors were evaluated for size, vascularity, location, and echogenecity (relative to placenta). Fetuses were assessed anatomically and for early signs of hydrops. Sonographic fetal and tumor examination were repeated weekly until delivery, and associated maternal complications were recorded. RESULTS: The sonographic diagnosis of chorioangiomas was subsequently confirmed pathologically in all cases. The maximum diameter of the tumors varied between 6 and 12 cm. Chorioangioma was hyperechogenic, avascular and complicated by intrauterine fetal growth restriction and oligohydramnios in one woman; a healthy but small for date infant was delivered at 38 weeks gestation. Two tumors were partially vascular and hyperechogenic, these tumors were associated with moderate polyhydramnios and preterm delivery of normal infants at 33 and 36 weeks. The remaining 3 tumors were hypoechogenic, diffusely vascular and were associated with severe polyhydramnios. In one case, the tumor was complicated by abrupt generalized non immune fetal hydrops and stillbirth; in the 2nd case, there was acute fetal distress and delivery of a premature anemic infant at 28 weeks; however, in the 3rd case, while the polyhydramnios and tumor vascularization decreased, its echogenecity increased and delivery of normal infant at full term was achieved. CONCLUSION: Sonographic assessment of echogenicity and vascularity of the large chorioangioma appears to be detrimental in predicting the clinical outcome of pregnancy. Spontaneous regression of tumor vascularity with subsequent resolution of hydramnios may occur. While vascular and hypoechogenic tumors are associated with higher incidence of pregnancy complications, favorable outcome is expected in avascular and hyperechogenic tumors. PMID- 11479639 TI - Pregnancy outcome in connective tissue diseases. AB - OBJECTIVE: To describe the pregnancy outcome in women with systemic lupus erythematosus and other connective tissue diseases attending a specialized high risk clinic at the Royal London Hospital. METHODS: Retrospective review of 30 patients seen over a 30-month period. Diagnoses included 18 patients with lupus and 5 with mixed connective tissue disease. Patients were evaluated following a standardized protocol. RESULTS: For all patients there were 28 (93%) live births and 2 spontaneous abortions. Of the live births, 6 (21%) were preterms and 5 (17%) were small for gestational age. For the 18-lupus patients there were 17 (94%) live births and one spontaneous abortion. Of the live births, 3 (18%) were preterm, 25% were small for gestational age and one had neonatal lupus erythematosus. Nine patients (30%) were positive for anti Sjogren's syndrome A antibodies: 7 systemic lupus erythematosus patients (39%) and 2 mixed connective tissue disease patients. Eight patients were positive for anti-phospholipid antibodies: 5 for anti-cardiolipin and 3 patients for lupus anticoagulant. CONCLUSION: Most pregnancies complicated by lupus do well with no maternal or neonatal deaths in a multidisciplinary high-risk clinic. However a high rate of prematurity and small gestational age remains a problem. PMID- 11479640 TI - Characteristics of rheumatoid arthritis relative to HLA-DR in Saudi Arabia. AB - OBJECTIVE: To determine the clinical characteristics of rheumatoid arthritis in Saudi Arabia in relation to human leukocyte antigen type. METHODS: A group of 91 rheumatoid arthritis patients, 72 females and 19 males were studied for the various clinical, laboratory and radiological parameters along with human leukocyte antigen-DR phenotypes. Since human leukocyte antigen-DR10 was most commonly associated with rheumatoid arthritis in our population, we compared those patients with human leukocyte antigen-DR10 to those without. RESULTS: The comparison yielded differences in the presence of rheumatoid nodules, erosions, corticosteroid treatment, joint involvement at presentation, hemoglobin levels, and white cell count. Only the last 3 parameters showed a statistical significance. CONCLUSION: Human leukocyte antigen type of Saudi patients with rheumatoid arthritis influenced the course of the disease but only to a limited extent. PMID- 11479641 TI - Etiologies of the urinary tract infections in a Yemeni City. AB - OBJECTIVE: To determine the causative agents of urinary tract infection in Yemeni patients in Sana'a city, and to determine the antibiotic susceptibilities of these agents in vitro. METHODS: Consecutive mid-stream urine specimens were submitted to our laboratory by 4029 patients in Sana'a city from 1990 to the end of 1999. The specimens were cultured and the isolates were identified using standard microbiological techniques. The antibiotic susceptibilities of the isolates were also determined (in vitro). RESULTS: The number of patients with urinary tract infection who yielded positive cultures from their mid stream urine specimens was 685 out of 4029 patients (17%). The ratio of female-to-male patients was 462:223 (2.07:1). The causative agents were mainly members of the family Enterobacteriaceae, 579 out of the 685 isolates (84.5%). Escherichia coli was the main pathogen, 357 isolates out of all the isolates (52%) followed by Klebsiella pneumoniae, 143 isolates (21%) and Proteus species, 56 isolates (8%). The other pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Enterobacter aerogenes, Staphylococcus saprophyticus, Corynebacterium urealyticum and Enterococcus faecalis) constituted 129 out of the 685 isolates (19%). There was a shift in the types of the causative agents of urinary tract infection between the first and 2nd halves of the study period. The isolation of Escherichia coli decreased from 60% in the first half to 42% in the 2nd. Whereas Klebsiella pneumoniae increased from 16% in the first half to 27% in the 2nd, Proteus species from 8% to 9%, Staphylococcus aureus from 5% to 7%, Pseudomonas aeruginosa from 3% to 7%, Enterobacter aerogenes from 3% to 3.5%, Staphylococcus saprophyticus from 2% to 3% and Corynebacterium urealyticum from 1% to 2%. Ninety percent of all the urinary pathogens were sensitive to ciprofloxacin, 75% to nitrofurantoin, 57% to gentamicin, 46% to cefaclor, 46% to nalidixic acid, 39% to doxycycline, 37% to co-trimoxazole and 20% to ampiclox. CONCLUSION: The types of urinary tract pathogens and their antibiotic susceptibilities in addition to the rate of isolation from male and female patients are reported. It is recommended that ciprofloxacin and nitrofurantoin or both are used in the blind treatment of urinary tract infection while awaiting the culture and sensitivity results. It is also recommended to continuously monitor the pattern of urinary pathogens from the community to guide the blind treatment of patients in the future. PMID- 11479642 TI - Prevalence of hepatitis C virus antibodies among health care workers in Damascus, Syria. AB - OBJECTIVE: Health care workers exposed to accidental inoculation with infected blood represent a high risk group of acquiring hepatitis C virus infection. Hepatitis C virus is considered the most common parenterally transmitted pathogen to which needle-stick recipients are exposed. Therefore, the prevalence of hepatitis C virus antibodies among different health care workers was studied. METHODS: Antibodies of hepatitis C virus were studied by 3rd generation enzyme immunoassay. Hepatitis B surface antigen and antibodies to hepatitis B core were carried out using enzyme immunoassays. Liver enzymes (alanine aminotransferase, aspartate aminotransferase) and total bilirubin were measured using reagents on chemistry autoanalyzer. The studied group (189 members, aged 28.1+/-6.2 years, 86 males and 103 females) consisted of laboratory workers (65), hemodialysis staff (34), dentistry workers (24), surgery workers (35), and a 5th group contained other medical care workers (31) in Damascus, Syria. RESULTS: The prevalence of hepatitis C virus antibodies among health care workers was 3%. The positivity of anti-hepatitis C was 0% in the laboratory group, dentistry group, and surgery group. Whereas, it was 6% in the hemodialysis group, and 10% in the other medical workers group. The prevalence of hepatitis B surface antigen was 6% among health care workers in Damascus. Means of biochemical parameters were 23.2+/-15.9 U/L for alanine aminotransferase, 21.8+/-7.8 U/L for aspartate aminotransferase, and 0.58+/-0.35 mg/dl for total bilirubin. CONCLUSION: The prevalence of hepatitis C virus antibodies among health care workers was higher than the prevalence among the general population (1%) which was determined during our study. However, the prevalence of anti-hepatitis C virus is lower than the prevalence of hepatitis B surface antigen among health care workers. There was no significant difference between means of biochemical parameters in each health workers and the general population. Factors related to infrastructure and operational system might be responsible for such prevalence of anti-hepatitis C virus among health care workers. An intensive periodic educational program for the medical and paramedical staff is important, in order to minimize the prevalence of anti hepatitis C virus among this important high risk group. PMID- 11479643 TI - Tetanus. A clinical review. AB - OBJECTIVE: Although tetanus is now uncommon in Oman (The Expanded Program on Immunization was introduced in 1981), patients continue to present from time to time at an average rate of 6 cases per year. Worldwide, the mortality rate for tetanus remains high (ranging from 15-20% in developed countries). At the Sultan Qaboos University Hospital, Muscat, Oman, prolonged Intensive Care Unit treatment and multi-disciplinary management is invariably required for tetanus patients. This study was carried out to evaluate our results over the past decade. METHODS: All tetanus patients admitted to the Sultan Qaboos University Hospital from 1991 up to the end of 1999 were retrospectively reviewed. Patients were diagnosed early, and aggressive treatment in the Intensive Care Unit was instituted, with a coordinated multidisciplinary management. RESULTS: Ten cases were identified and included in the review, comprising 9 adults and one infant aged 2 weeks. Adult patients were aged 36-75 years (mean 59 years), and the average Intensive Care Unit stay of the 9 surviving patients was 5.5 weeks (range 3-7 weeks). All patients presented with severe generalized tetanus. Two patients with traditional cautery marks developed tetanus. A focus of infection could not be found in 2 patients. All patients had early tracheostomy and assisted ventilation with appropriate sedation. One patient required almost 45 gm of diazepam throughout his Intensive Care Unit stay. One adult patient died on the 6th day of admission following myocardial infarction. The neonatal case survived after 35 days care in the Intensive Care Unit. The mortality rate for our patients was therefore 10%. CONCLUSION: Tetanus in Oman remains an infrequent but important disease requiring costly and prolonged Intensive Care Unit treatment. We attribute the comparatively low mortality rate (10%) in this study, to early diagnosis, institution of aggressive treatment, good nursing as well as a well-coordinated multi-disciplinary management. PMID- 11479644 TI - Brucellosis in children of Dhofar Region, Oman. AB - OBJECTIVE: To study the epidemiological and clinical pattern of brucellosis in children of Dhofar and to ascertain the efficacy of a pre-determined antibiotic regimen to treat the disease. METHODS: The study was hospital based and was carried out prospectively for 3 years. All cases diagnosed to have brucellosis on clinical and serological basis were entered into the study. The epidemiological background and clinical presentations were analyzed and the clinical response to a combination of oral rifampicin and co-trimoxazole was evaluated. RESULTS: Three hundred and seventy five cases of brucellosis were eligible for the study. Ingestion of raw milk and its products were responsible for causation of the disease in 63% of cases. Eighty three per cent had direct contact with animals mainly cattle. A minority of 4.5% denied ingestion of raw milk or coming into direct contact with animals. Fever was the most common presenting feature at 91%. We identified 2 distinct groups of presentation: Seventy per cent of those who presented with arthritis belonged to the older age group (7.34 years, standard deviation 2.64). They did not have a systemic illness. The younger age group presented with severe systemic illness associated with severe leucopenia and thrombocytopenia. The clinical response to the combination of rifampicin and co trimoxazole was satisfactory in 90% of patients and 98% of brucella species isolated from the blood of patients were sensitive to both antibiotics used. CONCLUSION: Ingestion of infected milk and contact with infected animals are the main causes of human brucellosis, although aerial transmission from contaminated environmental soil could not be excluded. The main clinical presentation of brucellosis in children is fever but the skeletal manifestations of the disease are significant. The hematological manifestations of the disease in endemic areas deserve special attention. The combination of oral rifampicin and co-trimoxazole for 6 weeks is adequate to treat most cases of brucellosis in children. PMID- 11479645 TI - The impact of using intranasal splints on morbidity and prevalence of adhesions. AB - OBJECTIVE: To study the effect of using intranasal splints for prevention of adhesions and to assess the morbidity associated with their use. METHODS: A retrospective study based on 2 tertiary hospitals from 1988-1995. One hundred and fourteen patients were divided into 2 groups. Group one with splints and group 2 without. The splints were used for 10 days and nasal toilet was carried out twice. Score of adhesions, perforation, bleeding, pain, crusting, and septum position were recorded. RESULTS: Adhesions in both groups were almost the same as well as septum medialization and perforation scores. Pain and crusting were more common in the splinted group (p-value <0.001). CONCLUSION: The use of intranasal splints should be individualized. Nasal irrigation is of importance to prevent crusting. The morbidity associated with intranasal splints should be considered before use. PMID- 11479646 TI - Psychiatric referrals. In primary care and general hospitals in Qassim Region, Saudi Arabia. AB - OBJECTIVE: From different perspectives, psychiatric symptoms have special significance in psychiatry. This study comparatively describes the psychopathological symptoms as noted in primary care (402) and general hospital (138) referrals. METHODS: Five hundred and forty psychiatric referrals, retrieved randomly, were reviewed extensively for collecting relevant data. RESULTS: Both hospital and primary care referrals were observed to have a variety of psychological and somatic symptoms of variable frequencies, which were suggestive of several psychopathological domains. Functional psychotic (19.5% versus 10%), mood (27.5% versus 23%) and psychosomatic (7% versus 2%) symptoms were significantly noted in hospital referrals as compared to primary care referrals while the later were observed to have significantly more somatic (34.5% versus 22.5%) and neurological (8% versus 4%) symptoms. Only a small proportion of primary care referrals (33/402, 8%] have symptoms of childhood psychiatric disorders. CONCLUSION: Psychiatric symptomatology differs in primary care and general hospital referrals. Both the general practitioners and clinicians are expected to record psychiatric symptoms in a comprehensive manner. Hence, they need condensed training courses on psychiatric symptomatology. PMID- 11479647 TI - Patient expectation and satisfaction in different hospitals in Irbid, Jordan. AB - OBJECTIVE: This study attempts to identify factors contributing to patients' satisfaction and to examine the relationship between patient satisfaction and patient expectations. METHODS: The study sample consisted of 360 patients randomly drawn from the outpatients' practice of 2 health centers in Irbid, North Jordan; a university and governmental one. Patients' satisfaction was assessed using a self-administered patient satisfaction questionnaire. Patients' expectations were assessed by exposing patients to a series of video clips showing pre-tested patient provider encounters. RESULTS: On average, users of the Community Health Center had lower expectation levels and higher satisfaction means when compared to users of the University Health Center. The study results showed that patient satisfaction was mainly influenced by patient expectation of received care even after adjustment for socio-demographic variables. CONCLUSION: This study therefore, argues that while assessment of patient satisfaction is useful as a monitoring indicator for overall health care delivery performance, still interventions are required to improve the delivered care. There is a need to examine client expectations and tailor services accordingly since satisfaction measures can only diagnose a problem while expectation assessment can identify clients needs and thus program managers can better design health services delivery. PMID- 11479648 TI - Multicystic renal dysplasia. AB - Multicystic renal dysplasia, the most common form of cystic renal disease in the newborn period, is a clinically important consequence of abnormal nephrogenesis. It usually presents as an abdominal mass. The dysplasias are usually unilateral, but it can be bilateral, segmental or focal. The clinical presentation usually depends on the extent of the dysplastic involvement and the degree of the associated urinary obstruction. Here, we present a case of histologically multicystic renal dysplasia, which is ?bilateral. The left kidney showed typical radiological, gross and histopathological features of multicystic dysplasia, but the right kidney showed only radiological features of dysplastic cystic kidney. PMID- 11479649 TI - Magnetic resonance imaging of the bone marrow in 3 patients with aplastic anemia. AB - Bone marrow appearances in aplastic anemia are characterized by the abundance of fatty marrow that replaces normal functional marrow. The signal intensity of aplastic bone marrow in sagittal T1-weighted magnetic resonance images of the spine is bright, resembling that of subcutaneous fat and, in most cases, is not difficult to differentiate from normal age-related marrow changes. Three patients with aplastic anemia are described, and the correlation of magnetic resonance imaging of the spine with bone marrow trephine biopsy findings in these patients is portrayed. Magnetic resonance imaging is an accessible, non-invasive technique that allows sampling of a larger volume of bone marrow tissue and is especially useful in the detection of fatty marrow replacement of the normal functional marrow in aplastic anemia. PMID- 11479650 TI - Ileo-colic intussusception in a patient with non-Hodgkin's lymphoma. AB - The gastro-intestinal tract is a common site for non-Hodgkin's lymphoma and its involvement leads to a variety of clinical presentations. In adults, intussusception has rarely been associated with gastro-intestinal non-Hodgkin's lymphoma. When it occurs, computerized axial tomography scanning seems to be the diagnostic tool of choice. PMID- 11479651 TI - Prevalence of hepatitis B and C markers in patients on maintenance hemodialysis in Najran. PMID- 11479652 TI - Antibiotic treatment of acute appendicitis - initial observations. PMID- 11479653 TI - A double-blind evaluation of the activity of an anti-cellulite product containing retinol, caffeine, and ruscogenine by a combination of several non-invasive methods. AB - A double-blind, randomized, placebo-controlled study was conducted with 46 healthy female volunteers in order to test an anti-cellulite product containing retinol, caffeine and ruscogenine. An evaluation of different parameters related to cellulite appearance, i.e., the skin macrorelief, the dermal and hypodermal structures, the skin mechanical characteristics, and the cutaneous flowmetry was assessed using several non-invasive methods. This combination of different evaluation methods resulted in the demonstration of significant activity of the anti-cellulite product versus baseline and showed its superiority versus the placebo in skin macrorelief (decrease of the "orange peel" effect) and an increase in cutaneous microcirculation. By using a combination of methods, it was possible to detail the activity of an anti-cellulite product and to show superiority of the product in comparison with the placebo. PMID- 11479654 TI - Differential scanning calorimetry studies of sebum models. AB - Human sebum is a mixture of triglycerides, fatty acids, wax esters, squalene, cholesterol, and cholesterol esters. P. acnes, a bacterium that is normally found on the skin, hydrolyzes certain triglycerides to fatty acids, thereby changing the sebum composition. The objective of this study was to examine the physical state of a model sebum and the effect of variations in its composition on its physical properties including (a) the carbon chain length of the components, (b) the ratio of unsaturated to saturated components, and (c) the ratio of triglycerides to fatty acids. A model sebum mixture was prepared based on a composition reported in the literature and evaluated by differential scanning calorimetry (DSC). Since cholesterol and cholesterol esters contribute insignificantly to sebum composition, they were not included. Squalene was kept constant (13%), while the concentration of the rest of the components was varied. Variations of sebum were prepared by dissolving all components in a 3:1 chloroform-methanol mixture for uniformity. Subsequently the solvent was evaporated at room temperature. The samples were then analyzed using DSC. Four distinct endotherms (namely, Mp-1, Mp-2, Mp-3, and Mp-4) were observed between 50 degrees C and 100 degrees C. Mp-1 and Mp-2 occurred below 0 degrees C and were contributed by unsaturated components. Mp-3 and Mp-4, which represent the saturated components, occurred above 30 degrees C. Thus, at normal skin temperature (skin surface temperature is 32 degrees C), sebum contains both a solid and a liquid phase. All the transition temperatures increased with an increase in carbon chain length for the same ratio of unsaturation to saturation. A replacement of unsaturated components with corresponding saturated components led to a decrease in the transition temperatures for the former (Mp-1 and Mp-2) and an increase in the transition temperatures for the latter (Mp-3 and Mp-4). Replacement of triglycerides with corresponding fatty acids (mimicking the action of anaerobic bacteria) caused an increase in Mp-2 and a decrease in Mp-4. In all cases, the final melting temperature (Mp-4) was greater than the temperature of the human skin surface (32 degrees C); thus components contributing to these endotherms are still solids at skin temperature. All variations in the sebum model led to mixtures of solids and liquids at skin temperature. Considering a reduction in Mp-3 and/or Mp-4 to represent sebum "fluidization," it was achieved by a decrease in carbon chain length, an increase in unsaturation, or a substitution of triglycerides by corresponding fatty acids. Preferential enrichment with the saturated species will lead to enrichment of solids versus liquids in the sebum, presumably making it difficult for the liquid phase to dissolve the solids. It seems plausible that perturbation of the balance of solid and liquid components of sebum, such as by P. acnes action, may lead to blockage of the follicle. Future research will investigate strategies to dissolve and/or liquify the solid phase of sebum. PMID- 11479655 TI - Effect of formulation on the delivery and metabolism of alpha-tocopheryl acetate. AB - The effect of delivery system on the permeation and metabolism of alpha tocopheryl acetate (alpha-TAc) was studied in micro-Yucatan pig skin, which closely resembles human skin. Various alpha-tocopheryl acetate formulations, including a simple isopropyl myristate (IPM) solution, an o/w emulsion, microemulsions, which differed in their oily phase content, and alcoholic and hydroalcoholic gels were made. A suitable HPLC method was developed and validated to separate and quantify alpha-TAc and alpha-tocopherol (alpha-T). Dulbecco's modified phosphate-buffered saline with 3% bovine serum albumin (DMPBS-BSA 3%) served as the receptor media to ensure tissue viability and to maintain skin conditions. Finite doses (5 microl) of the formulations were applied to viable pig skin using a statistically approved randomized complete block design. Data were analyzed using Tukey's studentized range test, and interday variability was estimated using an F-test. About 70% of the active was recovered from the wash, representing the amount adhering to the surface of the skin. alpha-TAc underwent metabolism in pig skin to the active antioxidant, alpha-T. The identity of the HPLC peaks were confirmed by spiking studies using known standards. The extent of metabolism was found to be formulation-dependent. No alpha-T was, however, detected in the stratum corneum. A higher extent of metabolism was obtained for the IPM solution, a microemulsion containing IPM as the oily phase, and the hydroalcoholic gel, when calculated based on the percent of total alpha-TAc permeated in the viable skin. Metabolism occurred in pig skin to the extent of 15 20% in terms of the total amount of alpha-TAc permeated in the viable skin and stratum corneum. Thus the topical delivery and metabolism of alpha-TAc were found to be dependent on formulation. PMID- 11479656 TI - Development of a device to measure human hair luster. AB - Evaluating human hair luster is important in developing effective hair care products. Many methods of measuring human hair luster have been proposed, but most have major disadvantages: some require cutting the subject's hair, some utilize bulky equipment, and some take much time. A device that does not impose excessive burden on the subject and hair, but can easily and conveniently measure human hair luster, has not yet been developed. Overcoming the disadvantages of the traditional method, our new device can measure luster accurately without cutting the hair. Neither the subject's hairstyle nor its color influences the measurements. The device is small, and the time required for measurement is only 0.2 sec. The hair of 84 subjects was evaluated using this device, and there was a high correlation between the sensory score and the measurements obtained. PMID- 11479658 TI - Risk factors for parenteral intoxication by mercury from dental amalgam. PMID- 11479659 TI - Exposure to elemental mercury in urban workers and gold miners from the Tapajos Region, Para, Brazil. PMID- 11479660 TI - Effect of soil loading and soil sequestration on dermal bioavailability of polynuclear aromatic hydrocarbons. PMID- 11479661 TI - Environmental exposure to polycyclic aromatic hydrocarbons and total suspended particulates in a Taiwanese temple. PMID- 11479662 TI - Toxin production in batch cultures of freshwater cyanobacterium Microcystis aeruginosa. PMID- 11479663 TI - Acute toxicity of 12 herbicides to the green algae Chlorella pyrenoidosa and Scenedesmus obliquus. PMID- 11479664 TI - Phytotoxicity of low-molecular-weight phenols from olive mill waste waters. PMID- 11479665 TI - Rank ordering of macroinvertebrate species sensitivity to toxic compounds by comparison with that of Daphnia magna. PMID- 11479666 TI - Effect of triphenyltin on duckweed Lemna minor. PMID- 11479667 TI - Evaluation of PCDD/F toxicity in fish livers from Ya-Er Lake, China: chemical analysis compared with in vivo and in vitro EROD bioassays. PMID- 11479668 TI - Solid suspended particles affecting the quality of air in urban environments. PMID- 11479669 TI - Determination and estimation of aqueous solubilities and n-octanol/water partition coefficients for phenylacrylates. PMID- 11479670 TI - Mobility of the rare earth elements with acid rainwater leaching in the soil column. PMID- 11479671 TI - Water pollution in an urban Argentine river. PMID- 11479672 TI - Variation of heavy metal contents in frozen vegetable products. PMID- 11479673 TI - Accumulation of heavy metals in vegetables grown in an industrial area in relation to soil. PMID- 11479674 TI - Study on the adsorption of vanadium (V) with Scenedesmus obliquus. PMID- 11479675 TI - Bioaccumulation of lead in wildlife dependent on the contaminated environment of the Kafue flats. PMID- 11479676 TI - Pesticides and heavy metals in agricultural soil of Kanpur, India. PMID- 11479677 TI - Covariance of copper concentrations in lobsters and seawater. PMID- 11479678 TI - Alternative splicing of the human VEGFGR-3/FLT4 gene as a consequence of an integrated human endogenous retrovirus. AB - The vascular endothelial growth factor receptor 3 (VEGFR-3/FLT4) is a receptor tyrosine kinase that regulates angiogenesis and vasculogenesis in response to the binding of the ligands VEGF-C and VEGF-D. Mutations in VEGFR-3 have been identified in patients with primary lymphoedema. It has been noted previously that whilst in the mouse there is only a single Vegfr-3 transcript, in humans there are two transcripts of 5.8 and 4.5 kb, of which the shorter encodes a protein that lacks the C-terminal 65 amino acids. These two isoforms also differ in their biological activity. Analysis of the human VEGFR-3 cDNA and genomic sequence reveals that these two isoforms arise by alternative splicing of the terminal exons. The shorter transcript is generated by splicing into the long terminal repeat of a human endogenous retrovirus located between the last two exons, thus explaining the lack of the shorter transcript in the mouse. The retention of the retroviral sequences in the FLT4 locus suggests that this retrotransposition event has contributed significant additional function to this gene. This provides support for a role for integrated retroviruses in modulating gene activity and participating in evolutionary processes. PMID- 11479679 TI - The complete sequence of a brown algal mitochondrial genome, the ectocarpale Pylaiella littoralis (L.) Kjellm. AB - We describe here the complete sequence (58,507 bp) of the mitochondrial genome of the brown alga Pylaiella littoralis (Ectocarpales). This molecule displays an AT content of 62.0% and contains seventy-nine genes, most of them (73) encoded on one strand. They include the usual mitochondrial set of protist genes and a number of rarer genes. Among these, several ribosomal protein genes and the rn5 were identified. Twenty-four tRNA genes are present in this genome, insufficient to decode all genes. The other conspicuous features of this molecule are: a large (3018 nucleotides) in-frame insertion of unknown function in the cox2 gene; the presence of two different lineages of group II introns, including complete reverse transcriptase-like genes, one in the cox1 and the other in the rnl gene; the concomitant occurrence of a T7-like RNA polymerase and of several well conserved alpha-proteobacterial-type promoters; and a small nad11 gene, coding for the first domain only of this NADH dehydrogenase subunit. Altogether, the mitochondrial genome of P. littoralis exhibits both alpha-proteobacterial characteristics and evidences of the independent integration of several exogenous DNA fragments. PMID- 11479680 TI - The cytochrome b gene as a phylogenetic marker: the limits of resolution for analyzing relationships among cichlid fishes. AB - The mitochondrial cytochrome b (cyt-b) gene is widely used in systematic studies to resolve divergences at many taxonomic levels. The present study focuses mainly on the utility of cyt-b as a molecular marker for inferring phylogenetic relationship at various levels within the fish family Cichlidae. A total of 78 taxa were used in the present analysis, representing all the major groups in the family Cichlidae (72 taxa) and other families from the suborders Labroidei and Percoidei. Gene trees obtained from cyt-b are compared to a published total evidence tree derived from previous studies. Minimum evolution trees based on cyt b data resulted in topologies congruent with all previous analyses. Parsimony analyses downweighting transitions relative to transversions (ts1:tv4) or excluding transitions at third codon positions resulted in more robust bootstrap support for recognized clades than unweighted parsimony. Relative rate tests detected significantly long branches for some taxa (LB taxa) which were composed mainly by dwarf Neotropical cichlids. An improvement of the phylogenetic signal, as shown by the four-cluster likelihood mapping analysis, and higher bootstrap values were obtained by excluding LB taxa. Despite some limitations of cyt-b as a phylogenetic marker, this gene either alone or in combination with other data sets yields a tree that is in agreement with the well-established phylogeny of cichlid fish. PMID- 11479681 TI - Origin and evolution of GBV-C/hepatitis G virus and relationships with ancient human migrations. AB - The GB virus C/hepatitis G virus (GBV-C/HGV) is a newly identified human RNA virus, belonging to the Flaviviridae family. Persistent infection by GBV-C/HGV is common in humans, and genetically divergent isolates have been identified in different parts of the world. Due to the absence of a real pathogenic role of GBV C/HGV in liver disease and its extremely low mutation rate, this virus is a potential marker to trace prehistoric links between human populations. In this study, origin and evolution of GBV-C/HGV were examined using a set of fully sequenced strains of worldwide origin. A first phylogenetic analysis, addressed to the short (255 nucleotides) NS5A overlapping coding region by the neighbor joining method, suggested an ancient African origin of GBV-C/HGV. This notion was confirmed when the same analysis was applied to the genomic regions showing the lowest rate of synonymous substitutions, covering one-fourth (2184 nucleotides) of the total coding potential of the virus genome. By using a multivariate statistical method and extending the analysis to the complete coding region, fine details of the evolutionary history of GBV-C/HGV were further elucidated. By this approach, isolates from Southeast Asia appeared to be the most closely related to those of African origin, consistent with a major route of ancient human migrations from Africa to southeastern parts of the Asian continent. PMID- 11479682 TI - The association between HLA-A alleles and an Alu dimorphism near HLA-G. AB - The AluYb8 sequences are a subfamily of short interspersed Alu retroelements that have been amplified within the human genome during recent evolutionary time and are useful polymorphic markers for studies on the origin of human populations. We have identified a new member of the Yb8 subfamily, AluyHG, located between the HLA-H and -G genes and 88-kb telomeric of the highly polymorphic HLA-A gene within the alpha block of the major histocompatibility complex (MHC). The AluyHG element was characterised with a view to examining the association between AluyHG and HLA-A polymorphism and reconstructing the history of the MHC alpha block. A specific primer pair was designed for a simple PCR assay to detect the absence or presence (dimorphism) of the AluyHG element within the DNA samples prepared from a panel of 46 homozygous cell-lines containing complete or recombinant ancestral haplotypes (AH) of diverse ethnic origin and 92 Caucasoid and Asian subjects on which HLA-A typing was available. The AluyHG insertion was most strongly associated with HLA-A2 and, to a lesser degree with HLA-A1, -A3, -A11, and A-19. The gene frequency of the AluyHG insertion for 146 Caucasians and 94 Chinese-Han was 0.30 and 0.32 and there was no significant difference between the observed and expected frequencies. The results of the association studies and the phylogenetic analysis of HLA-A alleles suggest that the AluyHG sequence was integrated within the progenitor of HLA-A2, but has been transferred by recombination to other human ancestral populations. In this regard, the dimorphic AluyHG element is an important diagnostic marker for HLA association studies and could help in elucidating the evolution and functions of the MHC alpha block and polymorphism within and between ancestral haplotypes. PMID- 11479683 TI - Extensive ribosomal DNA genic variation in the columnar cactus Lophocereus. AB - Sequence analysis of the hypervariable internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) is commonly used to gain insights into plant and animal population structure and phylogeny. We characterized ITS1, ITS2, and the 5.8S coding region of 18 senita (Lophocereus) individuals from 12 different populations in Baja as well as from closely related cactus species. Analyses of multiple clones demonstrated extensive paralogy in the senita rDNA gene family. We identified at least two putatively non-recombining rDNA operons in senita as well as multiple paralogous sequences within each operon. Usage of PCR, reverse transcriptase (RT)-PCR, Southern blot, primary sequence analyses of the 18S rDNA gene, and secondary structure analyses of the 5.8S rRNA showed that one of the operons encodes rDNA pseudogenes in a low copy-number (Truncated), whereas the second operon encodes an expressed rRNA (Functional). Surprisingly, we found extensive paralogy not only in the ITS regions but also in the 5.8S coding regions in senita both within and between operons. Phylogenetic analyses suggest that the second rDNA operon originated prior to the divergence of Lophocereus. A significant (p < 0.05) divergence-rate acceleration was found in the Lophocereus 5.8S rDNA coding region in the Functional operon in comparison to Pereskiopsis porteri (Cactaceae) and Portulaca molokiniensis (Portulacaceae) with Silene dioica and Spinacia oleracea as the outgroups. PMID- 11479684 TI - Loss of the gene for the alpha subunit of ATP synthase (ATP5A1) from the W chromosome in the African grey parrot (Psittacus erithacus). AB - This study describes the results of an analysis using Southern blotting, the polymerase chain reaction, and sequencing which shows that the African grey parrot (Psittacus erithacus) lacks the W-chromosomal gene for the alpha subunit of mitochondrial ATP synthase (ATP5A1W). Additional evidence shows that in other psittacines a fragment of the ATP5A1W gene contains five times as many nonsynonymous nucleotide replacements as the homologous fragment of the Z gene. Therefore, whereas in these other psittacines the corresponding ATP5A1Z protein fragment is highly conserved and varies by only a few, moderately conservative amino acid substitutions, the homologous ATP5A1W fragments contain a considerable number of, sometimes highly nonconservative, amino acid replacements. In one of these species, the ringneck parakeet (Psittacula krameri), the ATP5A1W gene is present in an inactive form because of the presence of a nonsense codon. Other changes, possibly leading to an inactive ATP5A1W gene product, involve the substitution of arginine residues by cysteine in the ATP5A1W protein of the mitred conure (Aratinga mitrata) and the blue and gold macaw (Ara ararauna). The data suggest also that although the divergence of the psittacine ATP5A1W and ATP5A1Z genes preceded the origin of the psittacidae, this divergence occurred independently of a similar process in the myna (Gracula religiosa), the outgroup used in this study. PMID- 11479685 TI - The Old World sparrows (genus Passer) phylogeography and their relative abundance of nuclear mtDNA pseudogenes. AB - The phylogenetic relationships of genus Passer (Old World sparrows) have been studied with species covering their complete world living range. Mitochondrial (mt) cyt b genes and pseudogenes have been analyzed, the latter being strikingly abundant in genus Passer compared with other studied songbirds. The significance of these Passer pseudogenes is presently unclear. The mechanisms by which mt cyt b genes become pseudogenes after nuclear translocation are discussed together with their mode of evolution, i.e., transition/transversion mitochondrial ratio is decreased in the nucleus, as is the constraint for variability at the three codon positions. However, the skewed base composition according to codon position (in 1st position the percentage is very similar for the four bases, in 2nd position there are fewer percentage of A and G and more percentage of T, and in 3rd codon position fewer percentage of G and T and is very rich in A and C) is maintained in the translocated nuclear pseudogenes. Different nuclear internal mechanisms and/or selective pressures must exist for explaining this nuclear/mitochondrial differential DNA base evolutive variability. Also, the phylogenetic usefulness of pseudogenes for defining relationships between closely related lineages is stressed. The analyses suggest that the primitive genus Passer species comes from Africa, the Cape sparrow being the oldest: P. hispaniolensis italiae is more likely conspecific to P. domesticus than to P. hispaniolensis. Also, Passer species are not included within weavers or Estrildinae or Emberizinae, as previously suggested. European and American Emberizinae sparrows are closely related to each other and seem to be the earliest species that radiated among the studied songbirds (all in the Miocene Epoch). PMID- 11479686 TI - Phylogenetic analysis of viroid and viroid-like satellite RNAs from plants: a reassessment. AB - The proposed monophyletic origin of a group of subviral plant pathogens (viroids and viroid-like satellite RNAs), as well as the phylogenetic relationships and the resulting taxonomy of these entities, has been recently questioned. The criticism comes from the (apparent) lack of sequence similarity among these RNAs necessary to reliably infer a phylogeny. Here we show that, despite their low overall sequence similarity, a sequence alignment manually adjusted to take into account all the local similarities and the insertions/deletions and duplications/rearrangements described in the literature for viroids and viroid like satellite RNA, along with the use of an appropriate estimator of genetic distances, constitutes a data set suitable for a phylogenetic reconstruction. When the likelihood-mapping method was applied to this data set, the tree likeness obtained was higher than that corresponding to a sequence alignment that does not take into consideration the local similarities. In addition, bootstrap analysis also supports the major groups previously proposed and the reconstruction is consistent with the biological properties of this RNAs. PMID- 11479687 TI - A statistical view of genome transcription? PMID- 11479689 TI - The induction of carbon monoxide-mediated airway relaxation by PACAP 38 in isolated guinea pig airways. AB - Pituitary adenylate cyclase--activating peptide 38 (PACAP 38) displays several biologic activities relevant to obstructive airway disease. Carbon monoxide (CO) has recently emerged as a potent, endogenously produced mediator of bronchodilation. In this study, we have analyzed the occurrence of PACAP 38 and the corresponding occurrence of heme oxygenase (HO), the rate-limiting enzyme for CO production, in guinea pig trachea, using immunocytochemistry. We have also investigated whether the dilatory effects of PACAP 38 are dependent on CO, using an in vitro setup for tracheal studies. A moderate supply of PACAP-like immunoreactive nerve fibers was seen in association with tracheal smooth muscle. HO-like immunoreactivity was observed in the respiratory epithelium and in association with smooth muscle bundles. PACAP 38 induced a concentration dependent relaxation of precontracted tracheal segments. This dilation was nearly abolished after pretreatment with zincprotoporphyrine, an inhibitor of heme oxygenase. The same effect was accomplished with Rp-8Br-cyclicGMPS, an inhibitor of cyclicGMP, whereas the nitric oxide synthase inhibitor N(G)-monomethyl-L arginine had no effect on the PACAP 38--induced dilation. The presented data suggest that PACAP 38 can induce bronchodilation by means of a CO-dependent, cyclicGMP-related mechanism, thereby providing a link between neurotransmission and local CO release in the airway smooth muscle. PMID- 11479690 TI - Inhibitory role of eosinophils on cell surface plasmin generation by bronchial epithelial cells: inhibitory effects of transforming growth factor beta. AB - Eosinophilic bronchitis is an essential component of bronchial asthma, and eosinophils play an important role. We studied the effect of eosinophils on cell surface plasmin generation by bronchial epithelial cells, because plasmin is thought to be involved in bronchial tissue repair/remodeling by means of fibrinolysis and the activation of proteases such as matrix metalloproteases. Plasmin was generated from exogenous plasminogen on the cell surface of cultured bronchial epithelial cells, NCI-H292. Transforming growth factor beta (TGF-beta) treatment resulted in reduced cell surface plasmin generation and a large increase in plasminogen activator inhibitor-type 1 (PAI-1) antigen production in NCI-H292 cells, whereas no conspicuous effects were observed with IL-1 beta and TNF alpha treatment (regulators in pulmonary epithelial cells). On the other hand, this cell surface plasmin generation was reduced by co-incubation with Eol 1, an eosinophil cell line. The addition of TGF-beta antisense and anti-TGF-beta antibodies attenuated this adverse effect of Eol-1 cell co-incubation. These data suggest that eosinophils play an inhibitory role on cell surface plasmin generation by bronchial epithelial cells by means of the up-regulation of PAI-1 expression induced by TGF-beta. Therefore, the accumulation of eosinophils in bronchial walls is thought to be involved in bronchial tissue repair/remodeling in asthma through this protease network. PMID- 11479691 TI - Establishment of pulmonary alveolar type II cell line from p53-deficient mice. AB - We obtained a pulmonary alveolar type II epithelial cell line, MAC7, derived from the lung of p53-deficient mice (p53 -/-). When this cell line was passaged for long periods of time, the epithelial cells grew at a high rate for over 50th passage and never entered the non-growing senescent phase characteristic of the normal cells (p53 +/+). Each pulmonary epithelial outgrowth was characterized morphologically and immunocytochemically, and the growth and viability of the outgrowths were examined by MTT assay. Expression of surfactant-associated protein, SP-B, was detected, and transmission electron microscopy demonstrated a type II phenotype containing lamellar bodies and phospholipids. These findings indicate that MAC7 cells retain the morphological and physiological properties of alveolar type II epithelial cells. This cell line should be useful in experimental systems for studying lung pathology. PMID- 11479692 TI - Analysis of T cell receptor V(beta) gene expression and clonality in bronchoalveolar fluid lymphocytes from a patient with chronic eosinophilic pneumonitis. AB - Chronic eosinophilic pneumonitis (CEP) is characterized by longstanding respiratory symptoms accompanied by a massive pulmonary eosinophil infiltration. T lymphocytes in bronchoalveolar lavage (BAL) from patients with chronic eosinophilic pneumonitis are considered to recognize unknown antigens. To analyze the pathogenesis of CEP, we examined the T cell receptor (TCR) repertoire and T cell clonotype of BAL lymphocytes and peripheral blood lymphocytes (PBLs) in a 66 year-old woman patient with CEP. The expression of TCR BV gene was analyzed by the family PCR method using specific primers for 20 TCR BV genes and BC gene. The clonotype of BAL and peripheral T cells was examined by the PCR-single-strand conformation polymorphism (SSCP) method. Functional sequences of some T cell clones were also carried out. A TCR repertoire of BAL T cells was heterogeneous as well as PBLs. However, SSCP analysis showed that distinct T cell clonotypes were detected in BAL T cells, TCR BV3, BV4, BV6, BV8, BV9, BV14, and BV18 positive T cell clones especially, expanded clonally in BAL from the patient. Sequencing analysis showed that GVD, LGG, RDXS, and SSG amino acid sequence motif were found in the CDR3 in lung-specific T cells. BAL-specific T cell clones accumulated in the patient with CEP. Thus, we can conclude that BAL T cells are induced by the antigen-driven stimulation and these cells might play a crucial role in the generation of CEP. PMID- 11479693 TI - The arterial site of action of nitric oxide in the neonatal pig lung determined by microfocal angiography. AB - To determine the site of action of inhaled nitric oxide (iNO) in the newborn pig lung, lungs were isolated and perfused at constant flow for microfocal x-ray angiography. Measurements of pulmonary arterial diameters were made on arteries in the 100--2500 microm diameter range under control conditions, during vasoconstriction caused by hypoxia (decreasing PO(2) from approximately 120 to approximately 50 Torr), or N(omega)-nitro-L-arginine methylester (L-NAME 10(-4) M) administration, with or without vasodilation induced by iNO (40 ppm) or by the NO donor S-nitroso-N-acetylpenicillamine (SNAP 5 x 10(-6) M) given intravascularly. Hypoxia caused constriction only in smaller arteries whereas L NAME constricted arteries throughout the size range studied. iNO dilated the smaller arteries more than the larger arteries under all study conditions. SNAP was used to provide an intravascular source of NO for comparison to iNO. SNAP also dilated smaller arteries more than larger arteries, but it had a significantly greater effect on the large arteries than did iNO. This suggests that differential accessibility of the vascular smooth muscle to NO between sources, air and blood, is a factor in the diameter dependence of the responses. PMID- 11479694 TI - Evaluation of Cyfra 21-1: a potential tumor marker for non-small cell lung carcinomas. AB - Cyfra 21-1 is a tumor marker based on the determination of water-soluble cytokeratin 19 which is secreted by normal or malignantly transformed epithelial cells. It is suggested to be a valuable marker in patients with non-small cell lung carcinoma (NSCLC). A prospective clinical study was conducted to investigate the value of Cyfra 21-1 for diagnosis, determination of subtypes, staging, and evaluation of therapy response in patients with lung carcinoma (Ca). Sixty-nine patients (mean age: 60.9 +/- 9.2 years, M/F:12.8) treated between 1994 and 1998 inclusive, and 13 healthy smokers (mean age:50.9 +/- 4.8 years, M/F:1.6) constituted the study group and control group, respectively. Venous blood samples (10 ml) were obtained from all subjects. Posttreatment blood samples were also obtained from 14 NSCLC patients. Cyfra 21-1 levels (cutoff value 3.3 ng/ml) were determined by ELSA-Cyfra 21-1 kit (CIS bio international, France) through immunoradiometric assay (IRMA). Cyfra 21-1 levels did not differ between smoking and non-smoking subjects within each group (p > 0.05). Cyfra 21-1 was significantly elevated in lung Ca cases irrespective of the cell type (p < 0.05). It was significantly elevated in squamous cell and adenocarcinoma varieties with the most prominent elevation in squamous cell type (p < 0.05). In lung Ca, the specificity and sensitivity of Cyfra 21-1 was 92.3% and 52.2%, respectively. Sensitivity was 65.5% for NSCLC, 70.5% for squamous cell, and 45.5% for adenocarcinoma varieties, with highest sensitivity rates in Stage IIIA + IIIB (87.5%) and Stage IV (75%) of squamous cell lung Ca. Cyfra 21-1 level was significantly decreased after treatment in NSCLC patients (n 14) (p < 0.01). Cyfra 21-1 is a tumor marker that helps to establish the diagnosis and differentiation of cell type and evaluation of response to therapy in patients with NSCLC. PMID- 11479697 TI - Sulfonate-sulfur metabolism and its regulation in Escherichia coli. AB - In the absence of sulfate and cysteine, Escherichia coli can use aliphatic sulfonates as a source of sulfur for growth. Starvation for sulfate leads to the expression of the tauABCD and ssuEADCB genes. Each of these gene clusters encodes an ABC-type transport system required for uptake of aliphatic sulfonates and a desulfonation enzyme. The TauD protein is an alpha-ketoglutarate-dependent dioxygenase that preferentially liberates sulfite from taurine (2 aminoethanesulfonic acid). SsuD is a monooxygenase that catalyzes the oxygenolytic desulfonation of a range of aliphatic sulfonates other than taurine. Its cosubstrate is FMNH2, which is provided by SsuE, an NAD(P)H-dependent FMN reductase. In contrast to many other bacteria, E. coli is unable to grow with arylsulfonates or with sulfate esters as sulfur source. The tau and ssu systems thus provide all genes for the utilization of known organosulfur sources by this organism, except the as yet unidentified gene(s) that enable some E. coli strains to grow with methanesulfonate or cysteate as a sulfur source. Expression of the tau and ssu genes requires the LysR-type transcriptional regulatory proteins CysB and Cbl. Synthesis of Cbl itself is under control of the CysB protein, and the CysB protein may therefore be regarded as the master regulator for sulfur assimilation in E. coli, while the Cbl protein functions as an accessory element specific for utilization of sulfur from organosulfur sources. PMID- 11479698 TI - Pattern of cyanophycin accumulation in nitrogen-fixing and non-nitrogen-fixing cyanobacteria. AB - The temporal and spatial accumulation of cyanophycin was studied in two unicellular strains of cyanobacteria, the diazotrophic Cyanothece sp. strain ATCC 51142 and the non-diazotrophic Synechocystis sp. strain PCC 6803. Biochemistry and electron microscopy were used to monitor the dynamics of cyanophycin accumulation under nitrogen-sufficient and nitrogen-deficient conditions. In Cyanothece sp. ATCC 51142 grown under 12 h light/12 h dark nitrogen-fixing conditions, cyanophycin was temporally regulated relative to nitrogenase activity and accumulated in granules after nitrogenase activity commenced. Cyanophycin granules reached a maximum after the peak of nitrogenase activity and eventually were utilized completely. Knock-out mutants were constructed in Synechocystis sp. PCC 6803 cphA and cphB genes to analyze the function of these genes and cyanophycin accumulation under nitrogen-deficient growth conditions. The mutants grew under such conditions, but needed to degrade phycobilisomes as a nitrogen reserve. Granules could be seen in some wild-type cells after treatment with chloramphenicol, but were never found in Delta cphA and Delta cphB mutants. These results led to the conclusion that cyanophycin is temporally and spatially regulated in nitrogen-fixing strains such as Cyanothece sp. ATCC 51142 and represents a key nitrogen reserve in these organisms. However, cyanophycin appeared to play a less important role in the non-diazotrophic unicellular strains and phycobilisomes appeared to be the main nitrogen reserve. PMID- 11479699 TI - A novel system for heterologous expression of flavocytochrome c in phototrophic bacteria using the Allochromatium vinosum rbcA promoter. AB - Flavocytochrome c-sulfide dehydrogenase (FCSD), an enzyme that catalyzes the reversible conversion of sulfide to elemental sulfur in vitro, is common to bacteria that utilize reduced sulfur compounds as electron donors in the process of carbon dioxide fixation. FCSD is a heterodimer containing two different cofactors, a flavin (FAD) and one or two heme c groups, located on the separate protein subunits. Efforts to produce the holoproteins of the soluble Allochromatium vinosum FCSD and the membrane-bound Ectothiorhodospira vacuolata protein in Escherichia coli using several expression systems were unsuccessful. Although all systems used were able to export the recombinant FCSDs to the periplasm, the proteins did not incorporate heme. In order to develop a new expression system involving photosynthetic hosts (Rhodobacter capsulatus, Rhodobacter sphaeroides and Ect. vacuolata), plasmid mobilisation from E. coli donors was studied. In the search for efficient promoters for such hosts, a system was developed combining the broad-host-range plasmid pGV910 and the promoter of the A. vinosum RuBisCo gene, rbcA. Conjugation was used to enable transfer from the expression plasmid of E. coli into Rba. capsulatus, Rba. sphaeroides strains and into Ect. vacuolata. Both Rhodobacter hosts were able to transcribe the genes coding for FCSD from the rbcA promoter and to produce detectable amounts of recombinant FCSD holoprotein. Western blots showed that the best production was obtained from cells grown photosynthetically on malate or acetate with sulfide. This system may prove to be of general use for the production of recombinant c-type cytochromes in homologous or related host systems. PMID- 11479700 TI - Elucidation of the pathways of catabolic glutamate conversion in three thermophilic anaerobic bacteria. AB - The glutamate catabolism of three thermophilic syntrophic anaerobes was compared based on the combined use of [(13)C] glutamate NMR measurements and enzyme activity determinations. In some cases the uptake of intermediates from different pathways was studied. The three organisms, Caloramator coolhaasii, Thermanaerovibrio acidaminovorans and strain TGO, had a different stoichiometry of glutamate conversion and were dependent on the presence of a hydrogen scavenger (Methanobacterium thermoautotrophicum Z245) to a different degree for their growth. C. coolhaasii formed acetate, CO(2), NH(4)(+) and H(2) from glutamate. Acetate was found to be formed through the beta-methylaspartate pathway in pure culture as well as in coculture. T. acidaminovorans converted glutamate to acetate, propionate, CO(2), NH(4)(+) and H(2). Most likely, this organism uses the beta-methylaspartate pathway for acetate formation. Propionate formation occurred through a direct oxidation of glutamate via succinyl-CoA and methylmalonyl-CoA. The metabolism of T. acidaminovorans shifted in favour of propionate formation when grown in coculture with the methanogen, but this did not lead to the use of a different glutamate degradation pathway. Strain TGO, an obligate syntrophic glutamate-degrading organism, formed propionate, traces of succinate, CO(2), NH(4)(+) and H(2). Glutamate was converted to propionate oxidatively via the intermediates succinyl-CoA and methylmalonyl-CoA. A minor part of the succinyl-CoA was converted to succinate and excreted. PMID- 11479701 TI - Structures of coenzyme F(420) in Mycobacterium species. AB - The structure of coenzyme F(420) in Mycobacterium smegmatis was examined using proton NMR, amino acid analysis, and HPLC. The two major F(420) structures were shown to be composed of a chromophore identical to that of F(420) from Methanobacterium thermoautotrophicum, with a side chain of a ribityl residue, a lactyl residue and five or six glutamate groups (F(420)-5 and F(420)-6). Peptidase treatment studies suggested that L-glutamate groups are linked by gamma glutamyl bonds in the side chain. HPLC analysis indicated that Mycobacterium tuberculosis, Mycobacterium bovis BCG, and Mycobacterium fortuitum have F(420)-5 and F(420)-6 as the predominant structures, whereas Mycobacterium avium contains F(420)-5, F(420)-6 and F(420)-7 in significant amounts. 7,8-Didemethyl 8-hydroxy 5-deazariboflavin (FO), an intermediate in F(420) biosynthesis, accounted for about 1-7% of the total deazaflavin in cells. Peptidase treatment of F(420) created F(420) derivatives that may be useful for the assay of enzymes involved in F(420) biosynthesis. PMID- 11479702 TI - ROSE elements occur in disparate rhizobia and are functionally interchangeable between species. AB - Expression of at least ten genes in Bradyrhizobium japonicum, seven of which code for small heat shock proteins (sHsps), is under the control of ROSE (repression of heat shock gene expression). This negatively cis-acting DNA element confers temperature control to a sigma(70)-type promoter. Here, we show that ROSE elements are not restricted to B. japonicum but are also present in Bradyrhizobium sp. (Parasponia), Rhizobium sp. strain NGR234 and Mesorhizobium loti. An overall alignment of all ROSE sequences reveals a highly conserved and probably functionally important region towards the 3'-end of the element. Moreover, we provide genetic evidence for the previously proposed presence of multiple sHsps in these organisms. Primer-extension data of five newly identified ROSE-associated operons show that transcription is repressed at low temperatures and induced after a temperature upshift. Translational ROSE-hsp'-'lacZ fusions of Bradyrhizobium sp. (Parasponia) and Rhizobium sp. strain NGR234 integrated into the chromosome of B. japonicum were heat-responsive. The functionality of these heterologous ROSE elements hints at a common regulatory principle conserved in various rhizobia. PMID- 11479703 TI - Characterization of a spontaneous adhesion-defective mutant of Ruminococcus albus strain 20. AB - A spontaneous adhesion-defective mutant (mutant D5) of Ruminococcus albus strain 20 was isolated and compared to the parent to investigate the impact of the mutation on cellulolysis and to identify the adhesion mechanism of R. albus. The comparison of kinetics of cellulose degradation by strain 20 and mutant D5 showed that the mutation delayed and reduced bacterial growth on cellulose and cellulose degradation. These results were partly explained by a twofold lower cellulase activity in the mutant than in the parent. The glycocalyx of strain 20, observed by transmission electron microscopy, was large and homogeneous, and linked cells to cellulose. The mutant glycocalyx was aggregated at its periphery and cells attached loosely to cellulose. A glycoprotein of 25 kDa (GP25), present in the membrane fraction and the extracellular medium of strain 20, was not detected in the same fractions of mutant D5. Though glycoprotein GP25 did not bind to cellulose, it may be involved in adhesion as an intermediate component. Different cell-surface features of mutant D5 (cellulases, glycoprotein GP25, glycocalyx) were thus affected, any or all of which may be involved in its adhesion-defective phenotype. These results suggest that adhesion and cellulolysis are linked and that adhesion is a multifactorial phenomenon that involves at least the extracellular glycocalyx. PMID- 11479704 TI - Characterisation of the mob locus of Rhodobacter sphaeroides WS8: mobA is the only gene required for molybdopterin guanine dinucleotide synthesis. AB - The mob genes of several bacteria have been implicated in the conversion of molybdopterin to molybdopterin guanine dinucleotide. The mob locus of Rhodobacter sphaeroides WS8 comprises three genes, mobABC. Chromosomal in-frame deletions in each of the mob genes have been constructed. The mobA mutant strain has inactive DMSO reductase and periplasmic nitrate reductase activities (both molybdopterin guanine dinucleotide-requiring enzymes), but the activity of xanthine dehydrogenase, a molybdopterin enzyme, is unaffected. The inability of a mobA mutant to synthesise molybdopterin guanine dinucleotide is confirmed by analysis of cell extracts of the mobA strain for molybdenum cofactor forms following iodine oxidation. Mutations in mobB and mobC are not impaired for molybdoenzyme activities and accumulate wild-type levels of molybdopterin and molybdopterin guanine dinucleotide, indicating they are not compromised in molybdenum cofactor synthesis. In the mobA mutant strain, the inactive DMSO reductase is found in the periplasm, suggesting that molybdenum cofactor insertion is not necessarily a pre requisite for export. PMID- 11479705 TI - The role of amino acids in the regulation of hydrogen sulfide production during ultradian respiratory oscillation of Saccharomyces cerevisiae. AB - We previously demonstrated that periodic H2S production during aerobic continuous culture of Saccharomyces cerevisiae resulted in ultradian respiratory oscillation, and that H2S production was dependent on the activity of sulfate uptake and the level of sulfite. To investigate the mechanism of regulation of the sulfate assimilation pathway and of respiratory oscillation, several amino acids were pulse-injected into cultures during respiratory oscillation. Injection of sulfur amino acids or their derivatives perturbed respiratory oscillation, with changes in the H2S production profile. Four major regulators of H2S production in the sulfate assimilation pathway and respiratory oscillation were identified: (1) O-acetylhomoserine, not O-acetylserine, as a sulfide acceptor, (2) homoserine/threonine as a regulator of O-acetylhomoserine supply, (3) methionine/S-adenosyl methionine as a negative regulator of sulfate assimilation, and (4) cysteine (or its derivatives) as an essential regulator. The results obtained after the addition of DL-propargylglycine (5 microM and 100 microM) and cystathionine (50 microM) suggested that the intracellular cysteine level and cystathionine gamma-lyase, rather than methionine/S-adenosylmethionine, play an essential role in the regulation of sulfate assimilation and respiratory oscillation. Based on these results and those of our previous reports, we propose that periodic depletion of cysteine (or its derivatives), which is involved in the detoxification of toxic materials originating from respiration, causes periodic H2S production. PMID- 11479706 TI - Superoxide dismutase plays an important role in the survival of Lactobacillus sake upon exposure to elevated oxygen. AB - In this study, the responses of two Lactobacillus sake strains to elevated oxygen concentrations at 8 degrees C were investigated. L. sake DSM 6333 (L. sake(sens)), unlike L. sake NCFB 2813 (L. sake(ins)), showed a low growth rate in the presence of 90% O(2) and a rapid loss in viability shortly after entry into stationary phase. The steady-state cytosolic superoxide radical (O(2)(-)) concentration in L. sake(sens) was 0.134 microM and in the oxygen-insensitive mutant LSUV4 it was 0.013 microM. The nine- to ten-fold decrease in the rate of O(2)(-) elimination in L. sake(sens) indicates the significance of the O(2)(-) scavenging system in protecting against elevated O(2). The superoxide dismutase (SOD) activity was 10- to 20-fold higher in L. sake(ins) than in L. sake(sens), depending on the growth phase. An oxygen-insensitive mutant of L. sake(sens), designated as strain LSUV4, had a ten-fold higher SOD activity than the wild-type strain, which likely restored its oxygen tolerance. Damage to proteins in L. sake(sens) was evidenced by the increased protein carbonyl content and reduced activities of the [Fe-S]-cluster-containing enzymes fumarase and fumarate reductase. This study forms a physiological basis for understanding the significance of elevated oxygen stress as an additional method for inhibition of microbial growth in relation to food preservation. PMID- 11479707 TI - Ethanedisulfonate is degraded via sulfoacetaldehyde in Ralstonia sp. strain EDS1. AB - Aerobic enrichment cultures (11) yielded three cultures able to utilise ethane 1,2-disulfonate as sole source of carbon and energy in salts medium. Two pure cultures were obtained and we worked with strain EDS1, which was assigned to the genus Ralstonia on the basis of its 16S rDNA sequence and simple taxonomic tests. Strain EDS1 utilised at least seven alkane(di)sulfonates, ethane-1,2-disulfonate, taurine, isethionate, sulfoacetate, sulfoacetaldehyde and propane-1,3 disulfonate, as well as methanesulfonate and formate. Growth with ethanedisulfonate was concomitant with substrate disappearance and the formation of 2 mol sulfate per mol substrate. The growth yield, 7 g protein (mol C)(-1), indicated quantitative utilisation of the substrate. Ethanedisulfonate-dependent oxygen uptake of whole cells during growth rose to a maximum before the end of growth and then sank rapidly; this was interpreted as evidence for an inducible desulfonative oxygenase that was not active in cell extracts. Inducible sulfoacetaldehyde sulfo-lyase was detected at high activity. Inducible degradation of taurine or isethionate or sulfoacetate via sulfoacetaldehyde sulfo lyase is interpreted from the data. PMID- 11479708 TI - Regulation of maltose transport in Saccharomyces cerevisiae. AB - Solute transport in Saccharomyces cerevisiae can be regulated through mechanisms such as trans-inhibition and/or catabolite inactivation by nitrogen or carbon sources. Studies in hybrid membranes of S. cerevisiae suggested that the maltose transport system Mal61p is fully reversible and capable of catalyzing both influx and efflux transport. This conclusion has now been confirmed by studies in a S. cerevisiae strain lacking the maltase enzyme. Whole cells of this strain, wherein the orientation of the maltose transporter is fully preserved, catalyze fully reversible maltose transport. Catabolite inactivation of the maltose transporter Mal61p was studied in the presence and absence of maltose metabolism and by the use of different glucose analogues. Catabolite inactivation of Mal61p could be triggered by maltose, provided the sugar was metabolized, and the rate of inactivation correlated with the rate of maltose influx. We also show that 2 deoxyglucose, unlike 6-deoxyglucose, can trigger catabolite inactivation of the maltose transporter. This suggests a role for early glycolytic intermediates in catabolite inactivation of the Mal61 protein. However, there was no correlation between intracellular glucose-6-phosphate or ATP levels and the rate of catabolite inactivation of Mal61p. On the basis of their identification in cell extracts, we speculate that (dideoxy)-trehalose and/or (deoxy)-trehalose-6 phosphate trigger catabolite inactivation of the maltose transporter. PMID- 11479709 TI - Development of a homologous transformation system for the opportunistic human pathogen Aspergillus fumigatus based on the sC gene encoding ATP sulfurylase. AB - The development of a homologous transformation system for the opportunistic human pathogenic fungus Aspergillus fumigatus is described. The system is based on the sC gene encoding ATP sulfurylase. Several A. fumigatus sC mutant strains were readily isolated by strong selection for selenate resistance. The coding region plus upstream and downstream regulatory sequences of the A. fumigatus sC gene were cloned by inverse PCR and then sequenced. Sequencing of the sC cDNA revealed the presence of five introns located within the first half of the gene. The A. fumigatus sC gene encodes a protein of 574 amino acids which is highly similar to ATP sulfurylases from the filamentous fungal species Aspergillus nidulans, Aspergillus terreus and Penicillium chrysogenum. By contrast, ATP sulfurylases from the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe lack the C terminal adenosine-5'-phosphosulfate kinase-like domain present in the filamentous fungal orthologues. A 3.8-kb DNA fragment amplified by PCR and containing the sC gene plus 5' and 3' flanking regions was cloned into pUC19 to give the vector pSCFUM. Transformation of two different sC mutant isolates with the plasmid pSCFUM established the functionality of this new homologous transformation system. Molecular analysis of sC+ transformants showed that up to 44% of transformed clones contained one or more copies of the entire plasmid integrated at the sC locus. This result also demonstrates the utility of the sC marker for targeting specific genetic constructs to the A. fumigatus sC locus, facilitating studies of gene regulation and function. PMID- 11479710 TI - Induction of butane consumption in Pseudomonas butanovora. AB - The induction of the enzyme activities involved in butane metabolism in Pseudomonas butanovora was characterized. P. butanovora was grown on butane or its metabolites, both singly and in mixtures with other growth substrates. Cells grown in each of the butane metabolites readily consumed the growth substrate and downstream metabolites, but consumed the upstream butane metabolites more slowly. Upstream activities in the butane metabolism could be induced by downstream metabolites, but to much lower levels than with the primary substrate. The induction of butane oxidation was not repressed when P. butanovora was grown or incubated in a mixture of butane and 1-butanol, butyraldehyde or butyrate. However, no induction of butane consumption was observed in a mixture of butane and lactate, which is indicative of catabolite repression. In lactate-grown cells that were rid of the growth substrate and incubated with butane and acetylene (to inactivate newly formed butane monooxygenase), the consumption of butane, 1 butanol and butyraldehyde consumption was not induced. The overall results suggest an independent regulatory mechanism for each of the enzyme activities in butane metabolism. In addition, a low, constitutive butane oxidation was observed in cells grown on substrates other than butane metabolites. PMID- 11479711 TI - Characterization of stress-responsive genes, hrcA-grpE-dnaK-dnaJ, from phytopathogenic Xanthomonas campestris. AB - Sequencing of a 6.4-kb DNA fragment, cloned from the plant pathogenic bacterium Xanthomonas campestris pv. campestris 17 revealed five ORFs whose deduced amino acid sequences show strong similarities to the bacterial HrcA, GrpE, DnaK, DnaJ, and PdxK. The four heat shock genes are organized in the order hrcA-grpE-dnaK dnaJ, a genome organization found in many gram-positive bacteria, but only in one gram-negative species (Xylella fastidiosa). These observations suggest that the HrcA-CIRCE system, comprising at least four genes arranged in this order, already existed for the regulation of stress responses before bacteria diverged into gram negative and gram-positive groups. Primer-extension results suggested the presence of promoters at the regions upstream of grpE and dnaK. In the presence of stress, heat or ethanol (4%), the X. campestris pv. campestris 17 grpE and dnaK promoters were induced two- to three-fold over controls. Since the grpE and dnaK promoters possess E. coli sigma(32) promoter-like sequences, they are functional in E. coli, although at levels much lower than in X. campestris pv. campestris 17. Furthermore, expression of the X. campestris pv. campestris 17 dnaK promoter in E. coli was elevated by the cloned X. campestris sigma(32) gene, indicating that the cognate sigma(32) works more efficiently for the X. campestris promoters. PMID- 11479712 TI - (S)-1-phenylethanol dehydrogenase of Azoarcus sp. strain EbN1, an enzyme of anaerobic ethylbenzene catabolism. AB - The initial steps in the anaerobic oxidation of the aromatic hydrocarbon ethylbenzene by denitrifying bacteria are two sequential dehydrogenation reactions of ethylbenzene to (S)-1-phenylethanol and further to acetophenone. The enzyme catalysing the second oxidation step, (S)-1-phenylethanol dehydrogenase, was analysed in the denitrifying bacterium Azoarcus sp. strain EbN1. An NAD+ dependent 1-phenylethanol dehydrogenase for each of the enantiomers of 1 phenylethanol was identified in this bacterium; the two enzymes were induced under different growth conditions. (S)-1-phenylethanol dehydrogenase from ethylbenzene-grown cells was purified and biochemically characterised. The enzyme is a typical secondary alcohol dehydrogenase and consists of two subunits of 25.5 kDa. The enantioselective enzyme catalyses the oxidation of (S)-1-phenylethanol or the reduction of acetophenone and is inhibited by high concentrations of (R)-1 phenylethanol. The enzyme exhibits low apparent K(m) values for (S)-1 phenylethanol and acetophenone and is rather substrate-specific, using only a few chemically similar secondary alcohols, such as 1-phenylpropanol and isopropanol. PMID- 11479713 TI - The nir, nor, and nos denitrification genes are dispersed over the Bradyrhizobium japonicum chromosome. AB - Cleavage of genomic DNA from Bradyrhizobium japonicum strain 3I1b110 by the restriction enzymes PmeI, PacI, and SwaI has been used together with pulsed-field gel electrophoresis and Southern hybridization to locate the nirK, norCBQD, and nosRZDFYLX denitrification genes on the chromosomal map of B. japonicum strain 110spc4. Mutant strains GRK13, GRC131, and GRZ25 were obtained by insertion of plasmid pUC4-KIXX-aphII-PSP, which carries recognition sites for the enzymes PacI, PmeI and SwaI, into the B. japonicum 3I1b110 nirK, norC and nosZ genes, respectively. Restriction of strain 3I1b110 genomic DNA with PacI, PmeI and SwaI yielded three, five and nine fragments, respectively. Pulsed-field gel electrophoresis of restricted mutant DNAs resulted in an altered fragment pattern that allowed determination of the position of the selected genes. Complementary mapping data were obtained by hybridization using digoxigenin-labeled B. japonicum 3I1b110 nirK, norBQD and nosZD as gene probes. The nirK, norCBQD and nosRZDFYLX genes were located close to the groEL(2), cycH and cycVWX genes, respectively, on the strain 110spc4 genetic map. In contrast to other denitrifiers, B. japonicum 3I1b110 denitrification genes were dispersed over the entire chromosome. PMID- 11479714 TI - 2-Oxoglutarate transport system in Staphylococcus aureus. AB - 2-[(14)C]oxoglutarate uptake in resting cells of Staphylococcus aureus 17810S occurs via two kinetically different systems: (1) a secondary, electrogenic 2 oxoglutarate:H(+) symporter (K(m)=0.105 mM), energized by an electrochemical proton potential (Delta mu H(+)) that is generated by the oxidation of endogenous amino acids and sensitive to ionophores, and (2) a Delta mu H(+)-independent facilitated diffusion system (K(m)=1.31 mM). The 2-oxoglutarate transport system of S. aureus 17810S can be classified as a new member of the MHS (metabolite:H(+) symporter) family. This transporter takes up various dicarboxylic acids in the order of affinity: succinate = malate > fumarate > 2-oxoglutarate > glutamate. Energy conservation with 2-oxoglutarate was studied in starved cells of strain 17810S. Initial transport of 2-oxoglutarate in these cells is energized by Delta mu H(+) generated via hydrolysis of residual ATP. Subsequent oxidation of the accumulated 2-oxoglutarate generates Delta mu H(+) for further, autoenergized transport of this 2-oxoacid and also for Delta mu H(+)-linked resynthesis of ATP. In the cadmium-sensitive S. aureus 17810S, Cd(2+) accumulation strongly inhibits energy conservation with 2-oxoglutarate at the level of Delta mu H(+) generation, without direct blocking of the 2-oxoglutarate transport system or ATP synthase complex. In the cadmium-resistant S. aureus 17810R, Cd(2+) does not affect energy conservation due to its extrusion by the Cd(2+) efflux system (Cd(2+)-ATPase of P type), which prevents Cd(2+) accumulation. PMID- 11479715 TI - The methionine biosynthetic pathway from homoserine in Pseudomonas putida involves the metW, metX, metZ, metH and metE gene products. AB - Biosynthesis of methionine from homoserine in Pseudomonas putida takes place in three steps. The first step is the acylation of homoserine to yield an acyl-L homoserine. This reaction is catalyzed by the products of the metXW genes and is equivalent to the first step in enterobacteria, gram-positive bacteria and fungi, except that in these microorganisms the reaction is catalyzed by a single polypeptide (the product of the metA gene in Escherichia coli and the met5 gene product in Neurospora crassa). In Pseudomonas putida, as in gram-positive bacteria and certain fungi, the second and third steps are a direct sulfhydrylation that converts the O-acyl-L-homoserine into homocysteine and further methylation to yield methionine. The latter reaction can be mediated by either of the two methionine synthetases present in the cells. PMID- 11479716 TI - The role of the natural polyamine putrescine in defense against oxidative stress in Escherichia coli. AB - Putrescine up-regulated, in a concentration-dependent manner, the expression levels of the oxyR and katG genes of Escherichia coli cells exposed to hydrogen peroxide. Its stimulatory effect was more pronounced under conditions of strong oxidative stress. 1,4-Diamino-2-butanone, a specific inhibitor of putrescine synthesis, also inhibited oxyR expression under oxidative stress. When added to inhibited cells, putrescine relieved this inhibitory effect. Addition of putrescine to E. coli cultures exposed to oxidative stress led to increased cell survival. PMID- 11479717 TI - Enzyme histochemically detectable NAD(P)H oxidase in human placental trophoblasts: normal, preeclamptic, and fetal growth restriction-complicated pregnancy. AB - The purpose of the present study was to determine the subcellular localization of NAD(P)H oxidase, a reactive oxygen species (ROS)-producing enzyme, in the human placenta at various gestational ages. Ultrastructural enzyme histochemistry for NAD(P)H oxidase, using cerium as a capturing agent, was carried out. Placentas from patients with severe preeclampsia and patients who delivered infants with fetal growth restriction (FGR) were also studied. Electron-dense precipitates indicating NAD(P)H oxidase activity were visible in the microvillous membranes of the placentas, especially on the surface plasma membrane of the syncytiotrophoblast microvilli, after 25 weeks of gestation. The distribution pattern and enzyme intensities were apparently the same among normal, preeclamptic, and FGR placentas. Cytochemical control experiments ensured the specific detection of NAD(P)H oxidase activity. These observations indicated that syncytiotrophoblasts possessed NAD(P)H oxidase activity, and thus ROS-generating activity. Placental NAD(P)H oxidase may play a role in placental lipid peroxidation and the placental defense mechanism. PMID- 11479718 TI - Morphology of the mitochondria and endoplasmic reticula of chorion laeve cytotrophoblasts: their resemblance to villous syncytiotrophoblasts rather than villous cytotrophoblasts. AB - We examined the morphological features of the mitochondria and endoplasmic reticula of chorion laeve cytotrophoblasts from term human fetal membranes, and compared them with those of syncytiotrophoblasts and cytotrophoblasts from human placental villi. Ultrastructural enzyme histochemistry of cytochrome c oxidase and glucose-6-phosphatase were used as cytochemical markers for these intracellular organelles. Chorion laeve cytotrophoblasts possessed abundant endoplasmic reticula, and small mitochondria with a few cristae, which were characteristic of villous syncytiotrophoblasts rather than villous cytotrophoblasts. As for these organellar structures, statistical analysis confirmed similarities between chorion laeve cytotrophoblasts and villous syncytiotrophoblasts, but significant differences between laeve cytotrophoblasts and villous cytotrophoblasts. Though these two cytotrophoblasts originated from one common cell in early placental development, they exhibited quite different organellar morphology during placental/chorioamniotic differentiation. Considering previous data, we concluded that chorion laeve cytotrophoblasts were metabolically active cells, similar to villous syncytiotrophoblasts, performing many functions in fetal membrane physiology. PMID- 11479719 TI - Immunolocalization of tubulin isoforms and post-translational modifications in the protists Tritrichomonas foetus and Trichomonas vaginalis. AB - In the present report we show the distribution of multiple tubulin isoforms in Trichomonas vaginalis and Tritrichomonas foetus, flagellated parasitic protists of the urogenital tracts of human and cattle, respectively, using immunofluorescence and immunoelectron microscopy. We used several monoclonal and polyclonal anti-tubulin antibodies from different sources and recognizing variant tubulin isoforms. Our results demonstrate that: (1) there is a heterogeneous distribution of the different tubulin isoforms in the main microtubular cell structures, such as axostyle, flagella, basal bodies, and mitotic spindle, (2) the axostyle-pelta junction is a structure with high affinity for glutamylated tubulin antibodies in T. foetus, (3) the spindle labeling is positive to anti glutamylated tubulin and anti-alpha-tubulin (TAT1 and purchased from Amersham) antibodies in T. vaginalis but it is negative in T. foetus, (4) the nuclear matrix and the cytosol presented positive reaction using glutamylated and TAT1 (anti-alpha-tubulin) antibodies only in T. vaginalis, and (5) the Golgi complex exhibited staining using the glutamylated tubulin antibody. The present data corroborate with the idea of the existence of a heterogeneous population of microtubules in these protists and of a subset of intracytoplasmic microtubules. Microtubule diversity may reflect distinct tubulins, diverse microtubule associated proteins, or a combination of both. PMID- 11479720 TI - Protein phosphatase 2Calpha expression in normal human tissues: an immunohistochemical study. AB - Protein phosphatase (PP2Calpha) is a member of the mammalian serine threonine specific protein phosphatases family. We produced monoclonal antibodies against the recombinant PP2Calpha and evaluated the immunoreactivity of normal human tissues. The reactivity was strong in normal skin, the digestive tract, lung, kidney, breast, prostate, endocrine glands, and brain, while it was moderate in the ovary, testis, and liver. Epithelial cells revealed high levels of PP2Calpha expression, but stromal cells, including fibroblasts and endothelial cells, showed no or little PP2Calpha expression. Given the broad reactivity in endocrine and secreting epithelial cells, we propose that PP2Calpha expression might contribute to secretory cell function. PMID- 11479721 TI - Recycling cultured cells for immunofluorescent labeling. AB - A method to use sequential rounds of immunofluorescent labeling in cell cultures is presented. The method is based on the utilization of a non-liquid reducing agent, sodium dithionite, in conjunction with ionic or non-ionic detergents (SDS or TX100, respectively) at room temperature. This method preserves cell morphology and substrate antigenicity, and operates through the complete extraction of most primary and secondary antibodies. Using this protocol, the sequential immunolocalization of different proteins is possible, without signal interference with previous immunolabeling rounds. In addition, the method is also useful to recycle blotted membranes in immunoblots. PMID- 11479722 TI - Developmental expression of CLC-K1 in the postnatal rat kidney. AB - CLC-K1, a kidney-specific chloride channel, has been demonstrated to be involved in the urine concentration mechanism. Here, we investigated the developmental expression of CLC-K1 in the rat kidney. Using immunohistochemistry, we showed that CLC-K1 was not present in the thin ascending limb of Henle's loop during the early prenatal stages but was significantly expressed during the adult stage. CLC K1 started to appear at day 5 and its expression increased during further development. In developing rats this increase coincided with the increase in the urine-concentrating capacity as the animals matured. We also investigated the expressions of other channels and transporters, including NKCC2, AQP-1, and AQP 2. NKCC2 was strongly expressed throughout the inner medulla in neonatal rat kidneys but was entirely undetectable at the adult stage. The decline in its expression took the form of a gradual recession from the inner medulla together with reciprocal increases in the expression of CLC-K1. AQP-1 was weakly expressed in the inner medulla during early development and showed a rapid increase in expression at a later stage. The collecting duct cells significantly expressed AQP-2 even at birth and maintained its expression throughout the development. These results suggest that CLC-K1 expression is one of the major determinants of the urine-concentrating capacity of the developing rat kidney. PMID- 11479723 TI - Cytokine expression in rat molar gingival periodontal tissues after topical application of lipopolysaccharide. AB - It is well known that proinflammatory cytokines produced by host cells play an important role in periodontal tissue destruction. However, the localization of the cytokines in in vivo periodontal tissues during development of periodontal disease has not been determined. Immunohistochemical expression of proinflammatory cytokines including IL-1alpha, IL-1beta, and TNF-alpha was examined at 1 and 3 h, and 1, 2, 3, and 7 days after topical application of lipopolysaccharide (LPS; 5 mg/ml in physiological saline) from E. coli into the rat molar gingival sulcus. In the normal periodontal tissues, a small number of cytokine-positive epithelial cells were seen in the junctional epithelium (JE), oral sulcular and oral gingival epithelium, in addition to macrophages infiltrating in the subjunctional epithelial area and osteoblasts lining the alveolar bone surface. Epithelial remnants of Malassez existing throughout periodontal ligament were intensely positive for IL-1beta but negative for the other two cytokines. At 3 h after the LPS treatment, almost all cells in the JE were strongly positive for the cytokines examined. In addition, several cytokine positive cells, including neutrophils, macrophages, and fibroblasts, were seen in the subjunctional epithelial connective tissue. At day 2, expression of the cytokines in the JE gradually decreased, while cytokine-positive cells in the connective tissue increased in number. Positive staining of the cytokines was seen in osteoclasts and preosteoclasts which appeared along the alveolar bone margin in this period. The number of cytokine-positive cells decreased by day 7. These findings indicate that, in addition to macrophages, neutrophils, and fibroblasts, the JE cells are a potent source of TNF-alpha, IL-1alpha, and IL 1beta reacting to LPS application, and suggest that JE cells may play an important role in the first line of defense against LPS challenge, and the proinflammatory cytokines transiently produced by various host cells may be involved in the initiation of inflammation and subsequent periodontal tissue destruction. PMID- 11479724 TI - Optimisation of oil red O staining permits combination with immunofluorescence and automated quantification of lipids. AB - The objective of the present study was to develop a stain permitting automated quantification of myocellular lipid depositions in skeletal muscle sections together with immunolocalisation of other myocellular constituents by fluorescence microscopy. Lipid droplets were detected in skeletal muscle by oil red O (ORO). Conventional ORO was modified to diminish background staining, prevent crystallisation of ORO and to optimise lipid retention in cryosections. These modifications resulted in a punctate staining of lipid droplets, rather than the somewhat diffuse staining by conventional ORO. Small cavities in muscle sections (like the lumen of small blood vessels) lack ORO when using the protocol presented here. In addition a staining protocol is presented combining ORO with immunofluorescence. This combination permits multiple staining studies in the same section. Thus, lipid droplets can be studied together with immunolabelling of proteins involved in lipid handling and metabolism. This will extend our knowledge on the subcellular localisation of lipid handling proteins (i.e. enzymes and fatty acid transporting proteins) in relation to the localisation of lipid depositions. In conclusion, the protocol presented here permits examination of ORO-stained lipid droplets in skeletal muscle sections together with multiple staining of other immunodetectable proteins present in skeletal muscle by quantitative fluorescence microscopy. PMID- 11479725 TI - Comparison of long-term transgene expression after non-viral and adenoviral gene transfer into primary articular chondrocytes. AB - Different gene transfer approaches to achieve long-term transgene expression in cultured primary bovine chondrocytes were compared using enhanced green fluorescent protein (EGFP) as a reporter. Transduction with a high-capacity adenoviral vector was 82% efficient when analysed by fluorescence microscopy, while up to 42% of plasmid-transfected cells were EGFP positive with FuGene as a transfection reagent. Rapid dominant marker selection of plasmid-transfected cells was achieved in monolayer culture. With either method of gene transfer, a high proportion of the chondrocytes remained transgene positive during prolonged alginate culture. Transgene transcription in single cells was quantified with a confocal laser scanning microscope. Detection of EGFP expression was more sensitive with this method, identifying more transgene-expressing cells than conventional fluorescence microscopy. Long-term EGFP expression was higher in adenovirally transduced chondrocytes embedded in alginate as compared to plasmid transfected cells cultured in monolayer or in alginate. Both the adenoviral and the plasmid-based approach appear suited for studies of the molecular and cellular mechanisms by which mutations in cartilage matrix proteins cause disease. PMID- 11479726 TI - Differentiation of the smooth muscle cell phenotypes during embryonic development of coronary vessels in the rat. AB - Smooth muscle cell (SMC) maturation during embryonic development of coronary arteries and veins was studied in rats using different markers of the contractile phenotypes. The spatio-temporal pattern of distribution of these markers compared with the developing tunica media was examined. Alpha-smooth muscle actin (alpha SMA) was the first marker of the SMC in the tunica media of coronary arteries found in ED16 hearts, followed by smooth muscle myosin heavy chain isoform which occurred on ED17. Subsequently 1E12 antigen was expressed in coronary artery wall in ED18 hearts, and finally smoothelin. The markers occur within the proximal part of the coronary arteries and deploy toward the apex. They are also found within the great vessels. None of the markers except for the alpha-SMA were found in coronary veins during embryonic life. We conclude that the SMC population of the developing tunica media of coronary vessels differentiates by the acquisition of particular markers and this process lasts till the end of the prenatal and early postnatal life. PMID- 11479727 TI - GFP associates with microfilaments in fixed cells. AB - The discovery of the green fluorescent protein (GFP) and its use as a marker for proteins in cells revolutionised cell biology. Among its applications are the intracellular localisation of proteins and the investigation of the organisation, regulation and dynamics of the cytoskeleton. GFP itself is considered to be an inert protein, homogeneously distributed within the cytoplasm. Here we investigated the intracellular distribution of GFP in an amphibian and in various mammalian cell lines (XTH2, CHO-K1, HaCaT, MDCK, NIH-3T3) by confocal laser scanning microscopy. After paraformaldehyde fixation GFP became associated with microfilaments in all the cell lines investigated. This interaction was not impaired by detergent treatment (1% Brij 58 for 10 min). In contrast to the F actin binding of GFP in fixed cells, association of GFP with stress fibres was not detectable in living cells. The actin-binding property of GFP might contribute also to the interaction of fusion proteins with microfilaments. Thus, careful controls are unavoidable in investigating (weak) actin-binding proteins in fixed cells. Because no association of GFP with microfilaments was detectable in living cells, it is recommended to monitor the intracellular distribution of GFP-tagged proteins in vivo. PMID- 11479728 TI - Identification of novel mutations in SHH and ZIC2 in a South American (ECLAMC) population with holoprosencephaly. AB - Holoprosencephaly (HPE) is genetically heterogeneous with four genes, SIX3, SHH, TGIF, and ZIC2 that have been identified to date and that are altered in 12% of patients. To analyze this prevalence in a South American population-based sample (57 HPE cases in 244,511 live and still births or 1 in 4300), we performed a mutational study of these genes in 30 unrelated children (26 newborns and 4 non newborns) with HPE being ascertained by ECLAMC (Latin American Collaborative Study of Congenital Malformations). We identified three novel mutations: two were missense mutations of the SHH gene (Cys183-->Phe; His140-->Pro); the third mutation was a 2-bp deletion in the zinc-finger region of the ZIC2 gene. These molecular results explained 8% (2/26 newborn samples) of the HPE cases in this South American population-based sample, a proportion similar to our previously published data from a collection of cases. PMID- 11479729 TI - Profiles of accepted mutation: from neutrality in a pseudogene to disease-causing mutation on its homologous gene. AB - We have compared the substitution pattern of the glucocerebrosidase gene (GBA) and the glucocerebrosidase pseudogene (psGBA), two highly homologous regions under different selective pressures and within the same genomic background. Mutations in GBA may lead to Gaucher disease, an inborn metabolic disorder. Disease-causing mutations and neutral variation in the gene have been compared to neutral variation in the pseudogene. This comparison offers a unique opportunity to better understand the action of purifying selection, since the differences between mutational patterns can be attributed to different selective pressures. A similar frequency of CpG dinucleotides was observed in GBA and in psGBA, and CpG pairs were mutated with the same high frequency in both regions. However, nucleotides not in CpG pairs were more likely to contribute to disease-causing mutation than to accepted polymorphisms. This pattern, which resulted in a lower transition to transversion ratio in the gene, may be due to CpG avoidance on critical regions within exons. PMID- 11479730 TI - Population-based risk estimates of Wilms tumor in sporadic aniridia. A comprehensive mutation screening procedure of PAX6 identifies 80% of mutations in aniridia. AB - Aniridia is a severe eye disease characterized by iris hypoplasia; both sporadic cases and familial cases with an autosomal dominant inheritance exist. Mutations in the PAX6 gene have been shown to be the genetic cause of the disease. Some of the sporadic cases are caused by large chromosomal deletions, some of which also include the Wilms tumor gene (WAGR syndrome), resulting in an increased risk of developing Wilms tumor. Based on the unique registration of both cancer and aniridia cases in Denmark, we have made the most accurate risk estimate to date for Wilms tumor in sporadic aniridia. We have found that patients with sporadic aniridia have a relative risk of 67 (confidence interval: 8.1-241) of developing Wilms tumor. Among patients investigated for mutations, Wilms tumor developed in only two patients out of 5 with the Wilms tumor gene (WT1) deleted. None of the patients with smaller chromosomal deletions or intragenic mutations were found to develop Wilms tumor. Our observations suggest a smaller risk for Wilms tumor than previous estimates, and that tumor development requires deletion of WT1. We report a strategy for the mutational analysis of aniridia cases resulting in the detection of mutations in 68% of sporadic cases and 89% of familial cases. We also report four novel mutations in PAX6, and furthermore, we have discovered a new alternatively spliced form of PAX6. PMID- 11479731 TI - Genetic basis of mitochondrial HMG-CoA synthase deficiency. AB - Deficiency of mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase (mHMGS) is a recessive disorder of ketogenesis that has been previously diagnosed in two children with hypoglycaemic hypoketotic coma during fasting periods. Here, we report the results of molecular investigations in a third patient affected by this disease. Sequencing of the entire coding region of the HMGCS2 gene revealed two missense mutations, G212R and R500H. Mendelian inheritance was confirmed by the analysis of parental samples and neither of the mutations was found on 200 control chromosomes. Functional relevance was confirmed by in vitro expression studies in cytosolic HMGS-deficient cells. Whereas wild-type cDNA of the HMGCS2 gene reverted the auxotrophy for mevalonate, the cDNAs of the mutants did not. The disease may be recognised by specific clinical and biochemical features but it is difficult to confirm enzymatically since the gene is expressed only in liver and testis. Molecular studies may facilitate or confirm future diagnoses in affected patients. PMID- 11479732 TI - Niemann-Pick type C disease: NPC1 mutations associated with severe and mild cellular cholesterol trafficking alterations. AB - Niemann-Pick type C disease (NPC) is a rare neurodegenerative disorder characterised by lysosomal/late endosomal accumulation of endocytosed unesterified cholesterol and delayed induction of cholesterol homeostatic reactions. The large majority of mutations in the NPC1 gene described thus far have been associated with severe cellular cholesterol trafficking impairment (classic biochemical phenotype, present in about 85% of NPC patients). In our population of 13 unrelated NP-C1 patients, among which 12 were of Portuguese extraction, we observed an unusually large proportion of families presenting mild alterations of intracellular cholesterol transport (variant biochemical phenotype), without strict correlation between the biochemical phenotype and the clinical expression of the disease. Mutational studies were carried out to compare molecular lesions associated with severe and mild cholesterol traffic impairment. Levels of NPC1 protein were studied by Western blot in cultured fibroblasts of four patients with homozygous mutant alleles. Ten novel mutations were identified (Q92R, C177Y, R518W, W942C, R978C, A1035V, 2129delA, 3662delT, IVS23+1 G>A and IVS16-82 G>A). The mutational profile appeared to be correlated with the biochemical phenotype. Splicing mutations, I1061T and A1035V, corresponded to "classic" alleles, while three missense mutations, C177Y, R978C and P1007A, could be defined as "variant" alleles. All "variant" mutations described so far appear to be clustered within the cysteine-rich luminal loop between TM 8 and 9, with the remarkable exception of C177Y. The latter mutant allele, at variance with P1007A, was correlated to a decreased level of NPC1 protein and a severe course of the disease, and disclosed a new location for "variant" mutations, the luminal loop located at the N-terminal end of the protein. PMID- 11479733 TI - Novel mtDNA mutations and oxidative phosphorylation dysfunction in Russian LHON families. AB - Leber's hereditary optic neuropathy (LHON) is characterized by maternally transmitted, bilateral, central vision loss in young adults. It is caused by mutations in the mitochondrial DNA (mtDNA) encoded genes that contribute polypeptides to NADH dehydrogenase or complex I. Four mtDNA variants, the nucleotide pair (np) 3460A, 11778A, 14484C, and 14459A mutations, are known as "primary" LHON mutations and are found in most, but not all, of the LHON families reported to date. Here, we report the extensive genetic and biochemical analysis of five Russian families from the Novosibirsk region of Siberia manifesting maternally transmitted optic atrophy consistent with LHON. Three of the five families harbor known LHON primary mutations. Complete sequence analysis of proband mtDNA in the other two families has revealed novel complex I mutations at nps 3635A and 4640C, respectively. These mutations are homoplasmic and have not been reported in the literature. Biochemical analysis of complex I in patient lymphoblasts and transmitochondrial cybrids demonstrated a respiration defect with complex-I-linked substrates, although the specific activity of complex I was not reduced. Overall, our data suggests that the spectrum of mtDNA mutations associated with LHON in Russia is similar to that in Europe and North America and that the np 3635A and 4640C mutations may be additional mtDNA complex I mutations contributing to LHON expression. PMID- 11479734 TI - Frequency of replication/transcription errors in (A)/(T) runs of human genes. AB - To estimate the error rate of the gene expression machinery and its possible age related increase, we compared the occurrence of polymerase errors during replication and transcription in (A)/(T) runs, in DNA and RNA of young and old individuals and of early- and late-passage cultured fibroblasts. We analyzed three human genes: TPRD, TGFBR2, and ATRX containing stretches of (A)8, (A)10, and (T)13, respectively. The error rate was determined by sequencing 100 cloned PCR or RT-PCR fragments from each DNA and RNA sample. The error rates in replication and transcription increased with the stretch length. The pooled error rates for genomic DNA were: TPRD (A)8, TGFBR2 (A)10, and ATRX (T)13: 1%+/-0.41, 15.8%+/-1.3, and 31.3%+/-2.9, while those for RNA were: 3.8%+/-0.5, 19.3%+/-2.1, and 54.3%+/-1.8, respectively. The deletions of one nucleotide were the most frequent errors. In the replication analysis, a significant difference was found in old versus young individuals for the ATRX (T)13. In the transcription analysis, significantly higher error rates were obtained in old versus young individuals for the TPRD (A)8 and TGFBR2 (A)10. For these genes, the error rate in RNA isolated from fibroblasts was significantly higher than that in blood. The data show a trend of age-related increase in replication/transcription errors; however further studies are necessary to confirm this hypothesis, since the sample size is small. This imperfect fidelity of the gene expression process may explain the evolutionary disadvantage of nucleotide repeats within coding sequences, and that these repeats are targets for mutations in human diseases. PMID- 11479735 TI - Identification of duplicated genes in 17q11.2 using FISH on stretched chromosomes and DNA fibers. AB - Recent evidence has been provided that links duplicons (REP-P and REP-M) in 17q11.2 flanking the neurofibromatosis type 1 (NF1) gene to the breakpoints of the NF1 microdeletion syndrome. The physical mapping and structural definition of duplicated regions is often impossible by conventional approaches, and so we have used high resolution fluorescence in situ hybridization (FISH) with locus specific probes of limited size on chromosomes stretched to different degrees to identify novel duplicated genes and expressed sequence tags (ESTs) mapping to 17q11.2. This approach has allowed us to detect and map duplications of the BLMH and GOS28 genes, with one copy lying centromeric and one telomeric to the NF1 gene, and an SHGC30113 transcript with one copy being adjacent and the other distal to the NF1 3' untranslated region. Double-color FISH with a BLMH-specific probe on stretched chromosomes showed that the telomeric BLMH repeat lacked the 5' end of the gene and was 0.8 Mb from its centromeric copy. The distance between the SHGC30113 repeats was estimated as being 500 kb by double-color FISH on highly stretched chromosomes and DNA fibers. The latter approach revealed adjacent SHGC30113-BLMH-specific signals relating to the incomplete BLMH copy. The use of FISH on stretched chromosomes and, where applicable DNA fibers, is a powerful tool for identifying and finely characterizing duplicated regions whose mapping by the classical physical mapping approach is often precluded. PMID- 11479736 TI - Glycerol kinase deficiency: evidence for complexity in a single gene disorder. AB - Glycerol kinase deficiency (GKD) occurs as part of an Xp21 contiguous gene syndrome or as isolated GKD. The isolated form can be either symptomatic with episodic metabolic and central nervous system (CNS) decompensation or asymptomatic with hyperglycerolemia and glyceroluria only. To better understand the pathogenesis of isolated GKD, we sought individuals with point mutations in the GK coding region and measured their GK enzyme activities. We identified six individuals with missense mutations: four (N288D, A305V, M428T, and Q438R) among males who were asymptomatic and two (D198G, R405Q) in individuals who were symptomatic. GK activity measured in lymphoblastoid cell lines or fibroblasts was similar for the symptomatic and the asymptomatic individuals. Mapping of the individuals' missense mutations to the three-dimensional structure of Escherichia coli GK revealed that the symptomatic individuals' mutations are in the same region as a subset of the mutations among the asymptomatic individuals, adjacent to the active-site cleft. We conclude that, like many other disorders, GK genotype does not predict GKD phenotype. We hypothesize that the phenotype of an individual with GKD is a complex trait influenced by additional, independently inherited genes. PMID- 11479737 TI - Chimpanzee apolipoprotein H (beta2-glycoprotein I): report on the gene structure, a common polymorphism, and a high prevalence of antiphospholipid antibodies. AB - Apolipoprotein H (apoH, protein; APOH, gene) is a 50-kDa glycoprotein that binds to negatively charged substrates, including phospholipids. ApoH is a main target antigen for the binding of antiphospholipid antibodies that are associated with thrombotic events. We have previously characterized the structural organization of the human APOH gene. Because of the significant structural homology between the human and chimpanzee genomes, we have employed oligonucleotides from the human APOH gene sequence to amplify chimpanzee DNA covering the entire transcribed region together with flanking sequence in the 5' region. As in humans, the chimpanzee APOH gene consists of eight exons and seven introns and encodes for a 326-amino-acid protein. The deduced amino acid and nucleotide sequence show 99.4% and 99.6% similarity between human and chimpanzee APOH, respectively. Using isoelectric focusing (IEF) and immunoblotting, we screened 155 chimpanzees (128 unrelated captured parents and 27 captive-born offspring) for the apoH protein polymorphism. The most common IEF pattern in chimpanzees was identical to a previously described APOH*3 allele in humans. In addition, an anodally shifted pattern was observed in chimpanzees with an allele frequency of 0.168, and the corresponding allele was designated as APOH*4. DNA sequencing of APOH*4 carriers revealed a missense mutation in exon 6 (A-->G) at codon 210, which replaces the amino acid lysine by glutamic acid. This mutation does not affect the binding of apoH to cardiolipin as revealed by cardiolipin/enzyme linked immunosorbent assay (ELISA). We also evaluated the prevalence of anti-apoH antibodies in chimpanzee plasma by using human-apoH-based ELISA and the association of the Lys210Glu mutation with the occurrence of anti-apoH antibodies. The prevalence of anti-apoH antibodies in chimpanzees (64%) was found to be unusually high compared with that found in humans. However, the Lys210Glu mutation showed no association with the occurrence of anti-apoH antibodies. The prevalence of anti-apoH antibodies in chimpanzees may serve as a useful animal model for the human antiphospholipid syndrome, where these antibodies are associated with clinical manifestations. PMID- 11479738 TI - Analysis of splicing parameters in the dystrophin gene: relevance for physiological and pathogenetic splicing mechanisms. AB - The molecular mechanisms that direct splice-site selection and assure orderly exon juxtaposition have not been fully clarified. The extraordinary nature of the dystrophin gene points to several hurdles in the processing of transcripts. In this study, dystrophin statistical and thermodynamic splicing parameters have been evaluated providing the first comprehensive description for a single human gene. We show that concomitant use of consensus values (CV) and DeltaDG degrees 37 values for U1snRNA annealing better discriminates between real donor sites and donor-like sequences. Evidence is also provided that, on average, out-of-frame dystrophin exons have significantly stronger CVs and more favorable DeltaDG degrees 37 values; this feature has never been reported and might reflect evolutionary-driven minimization of out-of-frame exon misplicing. Dystrophin splicing mutations have been reported to determine either Duchenne or Becker Muscular Dystrophy, but no comprehensive genotypic/phenotypic correlation has ever been investigated. We have analyzed splicing affecting single base-pair substitutions in the dystrophin gene with respect to their effect on splicing parameters; functional and clinical consequences are also reported. We have found 5'-splice-site mutation occurrence to be statistically related to mutability quotients and propose the use of DeltaDG degrees 37 values as a more effective tool than CV alone to describe donor site mutation consequences. Our analysis also indicates a nearly 100% correlation between clinical phenotype and the reading-frame rule determined at the RNA level. We consider that elucidation of the relative importance of splicing determinants might help to clarify the molecular mechanisms that direct correct splicing in complex genes and might be useful in the validation of predictive models. PMID- 11479739 TI - "Bar-coding" primate chromosomes: molecular cytogenetic screening for the ancestral hominoid karyotype. AB - Two recently introduced multicolor FISH approaches, cross-species color banding (also termed Rx-FISH) and multiplex FISH using painting probes derived from somatic cell hybrids retaining fragments of human chromosomes, were applied in a comparative molecular cytogenetic study of higher primates. We analyzed these "chromosome bar code" patterns to obtain an overview of chromosomal rearrangements that occurred during higher primate evolution. The objective was to reconstruct the ancestral genome organization of hominoids using the macaque as outgroup species. Approximately 160 individual and discernible molecular cytogenetic markers were assigned in these species. Resulting comparative maps allowed us to identify numerous intra-chromosomal rearrangements, to discriminate them from previous contradicting chromosome banding interpretations and to propose an ancestral karyotype for hominoids. From 25 different chromosome forms in an ancestral karyotype for all hominoids of 2N=48 we propose 21. Probes for chromosomes 2p, 4, 9 and Y were not informative in the present experiments. The orangutan karyotype was very similar to the proposed ancestral organization and conserved 19 of the 21 ancestral forms; thus most chromosomes were already present in early hominoid evolution, while African apes and human show various derived changes. PMID- 11479740 TI - A polymorphism in the promoter region of catalase is associated with blood pressure levels. AB - Catalase is an important antioxidant enzyme that detoxifies H2O2 into oxygen and water and thus limits the deleterious effects of reactive oxygen species (ROS). Because chronic exposure to excess ROS may contribute to vascular damage, we investigated whether genetic variation in catalase was associated with susceptibility to essential hypertension (EHYT) in 324 individuals (at least 50 years old) who were randomly sampled from an isolated population living in Xiangchang, China. They were screened for genetic variation in the promoter of catalase by direct sequencing. In total, four single nucleotide polymorphisms (SNPs) were identified. The association between the SNPs and EHYT was investigated by a linear regression model under phenotypic selection; in our analyses, we used both SBP>150 mmHg and SBP>160 mmHg as thresholds. A SNP 844 bp upstream of the start codon (SNP-844) demonstrated strong evidence of association with EHYT (SBP>150 mmHg: F=5.09, P=0.008; SBP>160 mmHg: F=7.13, P=0.002). This is the first study to implicate genetic variation in catalase in susceptibility to EHYT and suggests that polymorphisms in promoter regions may be particularly relevant to the study of complex diseases. PMID- 11479741 TI - A molecular approach to dominance in hypophosphatasia. AB - Hypophosphatasia is an inherited disorder characterized by defective bone mineralization and a deficiency of tissue-nonspecific alkaline phosphatase (TNSALP) activity. The disease is highly variable in its clinical expression, because of various mutations in the TNSALP gene. In approximately 14% of the patients tested in our laboratory, only one TNSALP gene mutation was found, despite exhaustive sequencing of the gene, suggesting that missing mutations are harbored in intron or regulatory sequences or that the disease is dominantly transmitted. The distinction between these two situations is of importance, especially in terms of genetic counseling, but dominance is sometimes difficult to conclusively determine by using familial analysis since expression of the disease may be highly variable, with parents of even severely affected children showing no or extremely mild symptoms of the disease. We report here the study of eight point mutations (G46 V, A99T, S164L, R167 W, R206 W, G232 V, N461I, I473F) found in patients with no other detectable mutation. Three of these mutations, G46 V, S164L, and I473F, have not previously been described. Pedigree and/or serum alkaline phosphatase data suggested possible dominant transmission in families with A99T, R167 W, and G232 V. By means of site-directed mutagenesis, transfections in COS-1 cells, and three-dimensional (3D) modeling, we evaluated the possible dominant effect of these eight mutations. The results showed that four of these mutations (G46 V, A99T, R167 W, and N461I) exhibited a negative dominant effect by inhibiting the enzymatic activity of the heterodimer, whereas the four others did not show such inhibition. Strong inhibition resulted in severe hypophosphatasia, whereas partial inhibition resulted in milder forms of the disease. Analysis of the 3D model of the enzyme showed that mutations exhibiting a dominant effect were clustered in two regions, viz., the active site and an area probably interacting with a region having a particular biological function such as dimerization, tetramerization, or membrane anchoring. PMID- 11479742 TI - Duplication and transposition of the NF1 pseudogene regions on chromosomes 2, 14, and 22. AB - Numerous NF1 pseudogenes have been identified in the human genome. Those in 2q21, 14q11, and 22q11 form a subset with a similar genomic organization and a high sequence homology. We have studied, by polymerase chain reaction and fluorescence in situ hybridization, the extent of homology of the regions surrounding these NF1 pseudogenes. Our analyses have demonstrated that a fragment of at least 640 kb is homologous between the three regions. Based on previous studies and these new findings, we propose a model for the spreading of the NF1 pseudogene containing regions. A fragment of approximately 640 kb was first duplicated in chromosome region 2q21 and transposed to 14q11. Subsequently, this fragment was duplicated in 14q11 and transposed to 22q11. A part of the 640-kb fragment in 14q11, with a length of about 430 kb, was further duplicated to a variable extent in 14q11. In addition, we have identified sequences that may facilitate the duplication and transposition of the 640-kb and 430-kb fragments. PMID- 11479743 TI - Galactosemia: deletion in the 5' upstream region of the GALT gene reduces promoter efficiency. AB - Galactosemia is a metabolic disorder caused by a defect in the galactose-1 phosphate uridyltransferase (GALT) enzyme. In previous studies, we have shown that the presence of a deletion in the 5' upstream (promoter) region of the GALT gene is associated with the Duarte (D2) allele. In the present study, by using a promoter fusion assay we provide direct evidence that a GTCA deletion located in position -119/-116 of the GALT gene (considered in relation to the translational start site) decreases transcription of a reporter gene to about 55% compared with a normal "healthy" promoter transfected into human hepatocyte HepG2 cells. This result coincides well with previously published biochemical data showing 50% GALT gene activity in Duarte (D2) galactosemia patients. By transfecting the same promoters (normal and deleted) into mouse NIH/3T3 cells, we show that the GTCA motif in the promoter region of the GALT gene was conserved throughout evolution. We conclude that the -119/-116delGTCA promoter mutation is a crucial factor in reduction of Duarte allele enzyme activity. PMID- 11479744 TI - Nomenclature for the description of human sequence variations. AB - A nomenclature system has recently been suggested for the description of changes (mutations and polymorphisms) in DNA and protein sequences. These nomenclature recommendations have now been largely accepted. However, current rules do not yet cover all types of mutations, nor do they cover more complex mutations. This document lists the existing recommendations and summarizes suggestions for the description of additional, more complex changes. Another version of this paper has been published in Hum Mut 15:7-12, 2000. PMID- 11479747 TI - MR arthrography for evaluation of the acetabular labrum. AB - MR arthrography of the hip has provided new insights into the lesions underlying mechanical hip pain. Until now evaluation of these structures, especially the acetabular labrum, has been limited. With the joint distention provided by MR arthrography, labral detachments and intrasubstance tears can be identified and differentiated from the many varied appearances of the asymptomatic labrum. This article reviews the normal anatomy of the hip, the clinical presentation of internal derangement and the technique for performing MR arthrography. The variations in morphology and signal of the asymptomatic labrum are reviewed as well as the appearance of the abnormal labrum. PMID- 11479748 TI - Destructive spondyloarthropathy of the cervical spine in long-term hemodialyzed patients: a five-year clinical radiological prospective study. AB - OBJECTIVE: To describe the radiographic features and progression of cervical spine destructive spondyloarthropathy (DSA) in hemodialyzed patients, and to evaluate the relationship between this disease and patient characteristics, biochemical values, and hemodialysis duration. DESIGN AND PATIENTS: Standard radiographs, and lateral flexion and extension views of the cervical spine, were performed annually for 5 years in 31 hemodialyzed patients who were divided into three groups at the commencement of the study: those showing (I) DSA, (II) vertebral rim erosions (VRE) without DSA, and (III) absence of DSA and VRE. Magnetic resonance (MR) imaging and computed tomography (CT) were performed in seven and two patients respectively. The imaging features were evaluated for the presence and progression of spondyloarthropathy and correlated with clinical and biochemical variables. Statistical analysis was performed using one-way analysis of variance. RESULTS: The duration of hemodialysis appeared to be the main predictive factor (P=0.0003) for DSA, which was found in six patients (19%). DSA was found to correlate with higher levels of beta2-microglobulin (P<0.00001), parathyroid hormone (P<0.05), and alkaline phosphatase (P<0.05). Clinical symptoms were minimal. In two patients, MR imaging revealed changes mimicking spondylodiscitis. In another patient, CT of the craniocervical junction showed bone resorption due to a pseudotumor, and basilar invagination. CONCLUSIONS: DSA of the cervical spine is often clinically silent. Pathogenesis of DSA may be multifactorial but its progression is most influenced by the duration of hemodialysis. On MR imaging, DSA may mimic spondylodiscitis. PMID- 11479749 TI - Benign vertebral hemangioma: MR-histological correlation. AB - OBJECTIVE: To explain the magnetic resonance (MR) appearance of benign vertebral hemangioma by correlating MR and histological findings from autopsy specimens. DESIGN: Sagittal T1- and T2-weighted spin-echo images were obtained in 83 spine specimens. Focal lesions consistent with vertebral hemangioma at macroscopic examination of sagittal anatomical sections were sampled for histological and quantitative analysis. At histology, the proportion of surface area occupied by adipocytes, vessels and edema, and hematopoietic cells was determined (point counting method) in normal marrow areas and in lesion areas whose signal intensity was either high and intermediate (pattern A) or intermediate and high (pattern B) on T1- and T2-weighted images, respectively. RESULTS: Nine lesions were sampled and corresponded to cavernous hemangioma at histology. The proportion of surface area occupied by adipocytes was statistically significantly higher in pattern A (78.1%) than in pattern B lesion areas (42.7%) and than in normal marrow areas (47.5%). The proportion of surface area occupied by vessels and interstitial edema was statistically significantly higher in pattern B (47.0%) than in pattern A lesion areas (15.5%) and than in normal marrow areas (0). CONCLUSION: The presence of high signal intensity on T1- or T2-weighted images of vertebral hemangioma is related to the amount of adipocytes or vessels and interstitial edema, respectively. PMID- 11479750 TI - Advanced imaging of melorheostosis with emphasis on MRI. AB - OBJECTIVE: To describe the CT and MR imaging appearance of both osseous and extraosseous manifestations of melorheostosis. DESIGN AND PATIENTS: We retrospectively reviewed the CT (n=7) and/or MR imaging findings (n=12) of 17 patients with characteristic radiographic findings of melorheostosis (undulating cortical hyperostosis with marked uptake on radionuclide bone scintigraphy). RESULTS: CT and MR imaging revealed cortical hyperostosis as high attenuation and low signal intensity on all MR pulse sequences, respectively. Encroachment on the marrow space was seen in all cases resulting from endosteal involvement. Thirteen patients demonstrated 14 soft tissue masses with infiltrative margins in 80% of cases and seven showed extensive mineralization on CT or MR imaging (low intensity on all pulse sequences). Seven soft tissue masses were predominantly nonmineralized with intermediate signal intensity on T1-weighted and intermediate to high signal on T2-weighted MR images corresponding to vascularized fibrous tissue with variable collagen content pathologically. Enhancement after intravenous gadolinium was seen in all patients imaged with soft tissue masses (n=2). Two patients demonstrated muscle atrophy resulting from nerve involvement. CONCLUSIONS: The osseous abnormalities in melorheostosis are identical on advanced imaging and radiographs. Mineralized or nonmineralized soft tissue masses should be recognized as another manifestation of this disease as opposed to a more ominous finding, making biopsy unwarrranted. PMID- 11479751 TI - Investigation of vertebral "end plate sclerosis". AB - OBJECTIVE: To evaluate the association between vertebral "end plate sclerosis" and neck pain. DESIGN: A retrospective study was carried out of lateral cervical spine radiographs with a Picture Archive and Communication System (PACS). PATIENTS: Two hundred patients' files were randomly assessed, comprising four equal groups, A to D. The mean ages of the patients were 62+/-7.4 years, 61+/-7.5 years, 40+/-5.6 years and 23+/-5.6 years respectively. In group A, all patients had symptoms of neck pain and a radiographic diagnosis of "end plate sclerosis" of the cervical spine. In groups B to D, asymptomatic patients were recruited and their age groups were 50-69, 30-49 and 10-29 years respectively. Using the PACS, the radiographic density and the sagittal diameter, thickness and area of the end plates at the C5 level were measured. RESULTS AND CONCLUSIONS: No significant differences were found in the radiographic density of the end plates either between the symptomatic and asymptomatic groups (groups A and B), or between different age groups (groups B, C and D). A significant increase in end plate area and thickness was found, however, in both group B (P<0.005) and group C (P<0.01) in comparison with group D. This indicates that the extent of end plate sclerosis increases with age. Our results suggest that the radiographic density of cervical vertebral end plates correlates neither with neck pain nor with increasing age. The radiological sign of "end plate sclerosis" may be over reported, further limiting its value in the assessment of patients with cervical spondylosis. PMID- 11479752 TI - Soft tissue twisting injuries of the knee. AB - OBJECTIVE: Twisting injuries occur as a result of differential motion of different tissue types in injuries with some rotational force. These injuries are well described in brain injuries but, to our knowledge, have not been described in the musculoskeletal literature. We correlated the clinical examination and MR findings of 20 patients with twisting injuries of the soft tissues around the knee. DESIGN AND PATIENTS: We prospectively followed the clinical courses of 20 patients with knee injuries who had clinical histories and MR findings to suggest twisting injuries of the subcutaneous tissues. Patients with associated internal derangement of the knee (i.e., meniscal tears, ligamentous or bone injuries) were excluded from this study. MR findings to suggest twisting injuries included linear areas of abnormal dark signal on T1-weighted sequences and abnormal bright signal on T2-weighted or short tau inversion recovery (STIR) sequences and/or signal to suggest hemorrhage within the subcutaneous tissues. These MR criteria were adapted from those established for indirect musculotendinous junction injuries. RESULTS: All 20 patients presented with considerable pain that suggested internal derangement on physical examination by the referring orthopedic surgeons. All presented with injuries associated with rotational force. The patients were placed on a course of protected weight-bearing of the affected extremity for 4 weeks. All patients had pain relief by clinical examination after this period of protected weight-bearing. CONCLUSIONS: Twisting injuries of the soft tissues can result in considerable pain that can be confused with internal derangement of the knee on physical examination. Soft tissue twisting injuries need to be recognized on MR examinations as they may be the cause of the patient's pain despite no MR evidence of internal derangement of the knee. The demonstration of soft tissue twisting injuries in a patient with severe knee pain but no documented internal derangement on MR examination may allow the orthopedic surgeon to elect for a trial of conservative nonsurgical management. PMID- 11479753 TI - Severe toxic osteoblastoma of the humerus associated with diffuse periostitis of multiple bones. AB - Toxic osteoblastoma is a rare variant of an uncommon primary bone tumour that has been described only once before in the world medical literature. It is characterised by systemic features including fever, weight loss and a striking diffuse periostitis in association with an osteoblastoma, resulting in delayed diagnosis. Early recognition of this condition could prevent inappropriate therapies and reduce morbidity. PMID- 11479754 TI - Pigmented villonodular synovitis of the hip presenting as a retroperitoneal mass. AB - We present an unusual case of diffuse pigmented villonodular synovitis (PVNS) of the hip presenting as a large retroperitoneal mass in an asymptomatic 39-year-old man. Initial imaging characteristics and findings on core needle biopsy suggested soft tissue sarcoma. However, incisional biopsy showed findings of PVNS. The patient underwent radical synovectomy and total joint replacement, with no change in the large retroperitoneal component after 15 months of follow-up. PMID- 11479755 TI - Synovial osteochondromatosis complicating pilon fracture of the tibia. AB - A case of previously undiagnosed synovial osteochondromatosis complicating a tibial pilon fracture is presented. The entrapment of osteochondral bodies within the fracture margin prevented complete reduction of the fracture and necessitated surgical intervention. PMID- 11479756 TI - Disproportionately severe calcinosis cutis in an 88-year-old patient with CREST syndrome. AB - An 88-year-old woman with CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasias) presented with hyperglycemia, intravascular depletion, and atrial fibrillation. The patient was found to have unusually severe calcinosis cutis in both legs extending from the knees to the ankles bilaterally, as well as Raynaud's phenomenon, sclerodactyly, and telangiectasias. The patient was normocalcemic and normophosphatemic. Although subcutaneous calcification is often seen with CREST syndrome, this case is unusual in that the area of involvement was much larger than previously described. Furthermore, the amount of calcinosis was disproportionately severe and was the major cause of symptoms and disability compared with the other components of the syndrome. PMID- 11479757 TI - Epidemiology of low back pain in the elderly: correlation with lumbar lordosis. AB - We carried out an epidemiologic study to determine the prevalence of low back pain in elderly Japanese and to examine the correlation with lumbar lordosis in sagittal plane radiographs. Low back pain is an enormous clinical and public health problem. With the increasing use of spinal instrumentation, the measurement of lumbar lordosis is thought to be important. However, in elderly Japanese, the prevalence of low back pain and its correlation with lumbar lordosis is not clear. Five hundred and nine people, aged 50-85 years, were examined, and 489 subjects met our criteria. Clinical findings, physical status, and the visual analogue scale (VAS) of pain were examined in these subjects. Measurements and determination of total lordosis from L1-S1 were made from standing radiographs. Forty-eight percent of the subjects had experienced low back pain within the previous 3 months. Women had low back pain more frequently (P = 0.006). There was a significant difference in lumbar lordosis between the groups with and without low back pain (P = 0.0006). Lumbar lordosis was approximately 4 degrees less in the low back pain group and there was no relationship to age or sex in either group. VAS was significantly inversely correlated with lumbar lordosis (P = 0.025, at rest). The body mass index (BMI) of the low back pain group was higher in women, but the difference was not significant (P = 0.06). In conclusion, lumbar lordosis was defined and its prevalence in elderly Japanese was reported together with VAS and physical data used to compare the two groups. PMID- 11479758 TI - The epidemiology of elbow fracture in children: analysis of 355 fractures, with special reference to supracondylar humerus fractures. AB - We present a study of the pattern of elbow fractures in children under 15 years of age, during a 5-year period, with special reference to supracondylar humerus fractures. The incidence was 308/100 000 per year; 58% of the children had a fracture in the supracondylar area of the humerus. There were 355 elbow fractures, and there were 164 boys (46%) and 191 girls (54%). The mean age for the entire group was 7.9 years (for boys, 7.2 years; for girls, 8.5 years). Of 209 supracondylar fractures (including 5 combination fractures), 134 were type I, 40 were type II, and 35 were type III (as classified by Gartland). Associated temporary nerve injuries involving the median, radial, and ulnar nerves were seen in 15 patients with type III supracondylar fractures. Associated brachial artery injuries were seen in 6 patients, 5 of whom had type III fractures. PMID- 11479759 TI - A new method of MR total spine imaging for showing the brace effect in scoliosis. AB - Bracing is a method of early, nonsurgical treatment for scoliosis, but a hypokyphotic effect on the thoracic spine is reported. We developed a magnetic resonance tomography (MR) procedure presenting an image of the whole spine in the coronal and sagittal planes (MR total spine imaging), and studied the brace effect, using this technique. We examined 26 female patients with idiopathic scoliosis treated with a Cheneau brace (mean age, 13.2 years; mean duration of brace treatment at the time of investigation, 1.5 years). The MR examinations were performed with the patient in the supine position with and without the brace in direct sequence. As measured on the coronal MR images, the thoracic curve was corrected, on average, from 29 degrees to 22 degrees (mean correction, 24%). There was a slight reduction in the sagittal Cobb angle measured between T4 and T12 (mean sagittal Cobb angle without brace, 14 degrees; with brace, 12 degrees ), which was still a significant change. MR total spine imaging could be a useful tool for studying the brace effect in scoliosis in two planes. Using this technique, we found reduced sagittal Cobb angles for the thoracic kyphosis with brace. Because there is no radiation exposure, the MR procedure has a potential use in the monitoring of brace treatment. PMID- 11479760 TI - Diagnosis of infection after total hip arthroplasty. AB - Forty-eight total hip arthroplasties for which revision surgery was performed were reviewed to determine the accuracy of laboratory tests, plain radiographs, hip aspiration, and technetium-99m MDP and gallium-67 scans in demonstrating the presence or absence of infection of the prosthesis. Six of the 48 hips were diagnosed as having an infection at the revision surgery. The erythrocyte sedimentation rate and the C-reactive protein levels were significantly higher in the patients with infected prostheses. The difference in the white blood cell count was not significant. There was no significant relationship between the presence of infection and the severity of loosening and instability of the implants diagnosed by plain radiographs. The accuracy of hip aspiration in diagnosing the infection was 83%, with a sensitivity of 40% and a specificity of 92%. The accuracy of technetium-99m MDP bone scan was 79%, with a sensitivity of 83%, and a specificity of 79%. Gallium-67 scan had an accuracy of 96%, a sensitivity of 67%, and a specificity of 100%. The findings in the present study indicated that diagnostic tests consisting of laboratory tests and plain radiography, followed by hip aspiration and sequential use of technetium-99m MDP and gallium-67 scintigraphies, are suitable for differentiation between mechanical loosening and infection of total hip arthroplasty. PMID- 11479761 TI - Minimum ten-year follow-up clinical and radiographic results of Mark I and Mark II Lord type femoral component in total hip arthroplasty. AB - The purpose of this study was to evaluate the clinical and radiographic results in 103 hips in 88 patients who had cementless primary Lord type femoral component (stem), with a mean follow-up of 11.8 years (range, 10 to 16 years). The original diagnoses were: osteoarthritis in 77 hips, rheumatoid arthritis in 15, and osteonecrosis of the femoral head in 11. Clinical results were assessed using the Harris hip score. Biological fixation, stem full-fit ratio in the intramedullary canal, and bone remodeling were evaluated by X-ray findings. The average clinical hip scores were: 47.1 points preoperatively, 81.7 points at 1 year, 86.6 points at 5 years, 79.0 points at 10 years, and 76.8 points at 15 years. By setting the endpoint at the time when loosening was radiologically confirmed, the 10-year survivorship of the femoral component was 96.9%, and the 16-year survivorship was 95.9%. The full-fit ratio between the bone implant interface just after surgery showed a significant relationship to stability of the femoral component. The decrease in the hip score at 10 years was caused by aseptic loosening of the acetabular threaded cup. However, the femoral component was stable for more than 10 years after the operation. In conclusion, long-term acceptable durability can be expected with the Lord type femoral component. PMID- 11479762 TI - Changes in knee motion following femoral and tibial lengthening using the Ilizarov apparatus: a cohort study. AB - We studied 46 patients undergoing tibial or femoral lengthening to correct limb length discrepancy (LLD) to assess the effects of the procedure on the range of motion (ROM) of the knee. The ROM of the knee has decreased even before the lengthening process started, and was at its most pronounced by the end of the lengthening. Mean preoperative ROM was 125 degrees +/- 21 degrees, and at follow up, at an average of 41 +/- 7.8 months from the beginning of the lengthening procedure, mean ROM was 117 degrees +/- 25 degrees (P = 0.31). There was no association between the worst ROM achieved during lengthening and the final ROM. Femoral lengthening resulted in a significantly greater loss of ROM for longer periods than tibial lengthening. Patients with congenital LLD were at risk of faster, more permanent loss of ROM than patients with posttraumatic or postinfective LLD. After the removal of the tibial or femoral Ilizarov frames, the ROM of the knee tended to return to prelengthening values. PMID- 11479763 TI - Discriminant analysis of prognostic factors for malignant fibrous histiocytoma in soft tissue. AB - We prospectively followed 32 patients with soft-tissue malignant fibrous histiocytoma (MFH). Parameters were age; sex; tumor size, location, and depth; operative method; chemotherapy; radiotherapy; and histology. Patients with recurrence or metastases due to MFH within 6 months after the initial operation were separated from those without these characteristics by discriminant analysis with statistically significant difference. The order of influential functions was histology, depth of tumor, operative method, and sex. Male patients with deep seated storiform-pleomorphic type MFH, receiving less comprehensive surgery, had the greatest risk of local recurrence or early metastases. We have to pay particular attention to patients with these factors and perform adequate surgery, because local recurrence and metastases were found to be closely related, and to have a great influence on the prognosis of this disease. Discriminant analysis to separate patients with MFH recurrence or metastases within 6 months after the initial operation from those without these characteristics is worthwhile for prognostic assessment. PMID- 11479764 TI - Anterior lumbar interbody fusion using two standard cylindrical threaded cages, a single mega-cage, or dual nested cages: a biomechanical comparison. AB - Our purpose was: (1) to compare the biomechanical properties of an interbody reconstruction using two standard threaded cages (18-mm diameter), a reconstruction using a single mega-cage (24-mm diameter), and a reconstruction using dual nested cages (22-mm diameter); and (2) to quantify the surface area of the cancellous bone bed exposed by reaming for the cages. Each motion segment was tested according to a nondestructive biomechanical loading sequence (compression, flexion, extension, lateral bending, axial torsion). Load was applied first to the intact motion segment and again after the insertion of cages, and stiffness values were calculated at each step. After the testing, each specimen was bisected through the disc and the surface area of the vascular bed was calculated. Comparison of the biomechanical properties of the three reconstructions showed that the dual nested cages produced the stiffest reconstruction. However, when the standard cages were compared with the nested cages, there was no significant difference, and compared with the mega-cage, the only difference was in flexion. The surface area of cancellous bone exposed by reaming for each of the three reconstructions showed the greatest value with the dual nested cages. These findings, together with the improved safety afforded by the nested or mega-cage, suggest that they are appropriate alternatives to the standard dual threaded cage reconstruction. PMID- 11479765 TI - The effects of prolonged peritendinous administration of PGE1 to the rat Achilles tendon: a possible animal model of chronic Achilles tendinopathy. AB - We studied the effects of peritendinous Achilles tendon injections of prostaglandin E1 (PGE1) on the Achilles tendon of rats. Five groups of Sprague Dawley rats (n = 24 each) were studied. Groups 1 to 4 received weekly peritendinous injections. In group 1, one side was injected with 800 ng of PGE1 in 0.5 ml of 0.9% NaCl and the contralateral side was injected with 0.5 ml of 0.9% NaCl. In group 2, one side was injected with 800 ng of PGE1. In group 3, one side was injected with 0.5 ml of 0.9% NaCl. In group 4, a syringe needle was inserted in the peritenon unilaterally, but no substances were administered. In groups 2, 3, and 4, the contralateral tendon was used as the control. In group 5, treatment was not administered. Eight rats in each group were killed at each time point, after 7, 21, and 35 days of treatment. On day 7, values for average water content and average wet weight of the tendons treated with PGE1 were significantly higher than those in the control tendons (analysis of variance [ANOVA]; P = 0.02), with a histological picture of acute inflammation. On day 21, approximately half of the PGE1-treated tendons showed fibrosis of the paratenon, with adhesions and intra-tendinous degeneration, with the other half still showing a picture of acute inflammation. On day 35, all of the PGE1-treated tendons showed fibrosis of the paratenon, with adhesions and intra-tendinous degeneration. At all time points, there was no evidence of pathology in the tendons that had not received PGE1. Sham peritendinous injections and injections of normal saline did not produce inflammation in the Achilles tendons. Initially, local administration of PGE1 produced acute inflammation of the tendon and its surrounding tissues. Prolonged PGE1 administration produced peri- and intra tendinous degeneration, providing a cheap, reproducible model of Achilles tendinopathy, which would allow studies of the effects of conservative and surgical management of the condition. PMID- 11479766 TI - Limb-lengthening and angular correction for dysplasia epiphysealis hemimelica. AB - We report a patient with severe varus deformity and tibial shortening caused by dysplasia epiphysealis hemimelica (DEH), a rare skeletal developmental disorder characterized by an osteocartilaginous tumor arising from an epiphysis in young children. Limb lengthening and angular correction was performed successfully, with an excellent result without abnormal bone growth. PMID- 11479767 TI - Transabdominal approach for intrapelvic migration of a total hip prosthesis component. AB - We report a patient with severe intrapelvic migration of the acetabular component after total hip arthroplasty. Preoperative drip infusion pyelography showed a medial shift in the ureter caused by the migrated acetabular component. Preoperative angiography showed that the right external iliac artery was compressed and shifted medially by the migrated acetabular component. At revision surgery, the migrated acetabular component was successfully removed through a transabdominal (transperitoneal) approach, with the assistance of a general surgeon, and reconstruction was performed through a transtrochanteric approach. We concluded that the transabdominal approach was a useful approach for removing a severely migrated acetabular component. PMID- 11479768 TI - Ewing's sarcoma: presentation and management. AB - Great strides have been made in the diagnosis and treatment of patients with Ewing's sarcoma. With the advent of modern chemotherapy, the long-term survival has improved to approximately 70%. Standard treatment for local control of the primary lesion has, historically, been chemotherapy and radiation. Currently, surgical resection has become a more effective option in the multidisciplinary treatment of patients with this disease. These current concepts and developments in the presentation and management of Ewing's sarcoma are discussed. PMID- 11479769 TI - The contributions of electron microscopy to the understanding and diagnosis of plasma cell dyscrasia-related renal lesions. AB - Electron microscopy has been crucial in the definition of many renal diseases. Ultrastructural evaluation has been instrumental in the characterization of many of the morphological manifestations of plasma cell dyscrasia-related renal lesions. Although it is recognized that there is controversy in regards to what the term multiple myeloma specifically refers, for the purposes of this article, myeloma and plasma cell dyscrasia are used interchangeably without consideration to perhaps significant conceptual differences that may exist between the two. Although distal nephron obstructive nephropathy ("myeloma kidney") was rather accurately defined solely on the basis of light microscopic findings and the association of AL-amyloidosis with underlying myeloma was confirmed using histochemical stains, the more subtle and intricate expressions of plasma cell dyscrasia-associated pathology required the electron microscope for proper characterization. The fibrillary nature of amyloid was discerned ultrastructurally, and detecting its characteristic ultrastructural features remains paramount to make a definitive diagnosis of amyloidosis when evaluating difficult cases. Pristine light- and heavy-chain deposits were objectively confirmed as immunomorphological correlates were depicted by immunofluorescence and at the ultrastructural level, substantiating the findings. Tubular interstitial alterations in these disorders other than cast nephropathy were firmly documented when careful ultrastructural studies were conducted experimentally and using clinical material. Likewise, electron microscopy has also played an important role in assessing vascular pathology in these conditions, especially when changes are focal, segmental, subtle, or such that they are easily confused with other pathological entities by light microscopy. Had it not been for the electron microscope, a clear definition of the immunomorphological scope of plasma cell dyscrasia-associated lesions, as understood currently, would not have been possible. Immunoelectron microscopy has provided a welcome added dimension, allowing a thorough characterization by expanding merely morphological data and providing exquisite immunomorphological correlations. This review highlights the role that electron microscopy has played and continues playing in the characterization of plasma cell dyscrasias-related renal lesions. PMID- 11479770 TI - The ultrastructural and immunohistochemical heterogeneity of CD-30-positive neoplasms: so-called anaplastic large cell Ki-1 lymphomas. AB - Anaplastic large cell lymphoma (ALCL), also referred to as Ki-1 lymphomas, was first recognized as an entity with characteristic light microscopic appearance in 1985. This tumor is composed of variably cohesive cells, often with large, markedly atypical, and multinucleated cellular forms. The recognition of ALCL resulted from the development of a monoclonal antibody in Kiel, Germany, named Ki 1, which was initially believed to be a putative marker for Reed-Sternberg cells. This antibody was later found to be specific against the epitope CD-30. Attempts to create strict criteria to preserve this neoplasm as a specific entity have undergone evolution. However, it is now clear that included in this group are a variety of pleomorphic neoplasms with CD-30 immunoreactivity. Some of these neoplasms are nonlymphoid and show marked heterogeneity in their immunohistochemical and ultrastructural profiles. This article aims to highlight the ultrastructural spectrum of neoplasms exhibiting CD-30 positivity that are within the spectrum of ALCL. It remains to be determined if there are subgroups of these CD-30-positive neoplasms that can be segregated on the basis of ultrastructural and immunohistochemical criteria with corresponding clinical correlates that may impact on their management, treatment, and prognosis. We review here the heterogeneity of CD-30-positive neoplasms (so-called anaplastic large cell Ki-1 lymphomas). PMID- 11479771 TI - Pathogenesis of acne. AB - Acne vulgaris is a skin disorder of the sebaceous follicles that commonly occurs in adolescence and in young adulthood. The major pathogenic factors involved are hyperkeratinization, obstruction of sebaceous follicles resulting from abnormal keratinization of the infundibular epithelium, stimulation of sebaceous gland secretion by androgens, and microbial colonization of pilosebaceous units by Propionibacterium acnes, which promotes perifollicular inflammation. The clinical presentation of acne can range from a mild comedonal form to severe inflammatory cystic acne of the face, chest, and back. At the ultrastructural level, follicular keratinocytes in comedones can be seen to possess increased numbers of desmosomes and tonofilaments, which result in ductal hypercornification. The increased activity of sebaceous glands elicited by androgen causes proliferation of P. acnes, an anaerobe present within the retained sebum in the pilosebaceous ducts. The organism possesses a ribosome-rich cytoplasm and a relatively thick cell wall, and produces several biologically active mediators that may contribute to inflammation, for instance, by promoting leukocyte migration and follicular rupture. In inflamed lesions, numerous neutrophils and macrophages infiltrate around hair follicles and sometimes phagocytose P. acnes. To examine the participation of neurogenic factors in the pathogenesis of acne, we quantitatively assessed the effects of neuropeptides on the morphology of sebaceous glands in vitro using electron microscopy. Substance P, which can be elicited by stress, promoted the development of cytoplasmic organelles in sebaceous cells, stimulated sebaceous germinative cells, and induced significant increases in the area of sebaceous glands. It also increased the size of individual sebaceous cells and the number of sebum vacuoles for each differentiated sebaceous cell, all of which suggests that substance P promotes both the proliferation and the differentiation of sebaceous glands. In this review, we introduce the general concept of pathogenic factors involved in acne, including typical electron microscopic findings and recent evidence of stress induced exacerbation of acne from a neurological point of view. An improved understanding of the pathogenesis of acne should lead to a rational therapy to successfully treat this skin disease. PMID- 11479772 TI - Expressions of endothelin-1, fibronectin, and interleukin-1alpha of human umbilical vein endothelial cells under prolonged culture. AB - We examined human umbilical vein endothelial cells (HUVECs) under prolonged culture by electron microscopy and by light and electron immunocytochemistry including double immunolabeling. Based on the cell area of HUVECs through multiple passages, we divided the cells into first, second, and third stages, which exhibited distinct morphological and immunocytochemical characteristics. During the first stage, HUVECs were polygonal in shape and had already formed the monolayer confluence. During the second stage, they were characterized by an increased number of Weibel-Palade (WP) bodies, which were actively segregated from Golgi cisterns. Endothelin (ET)-1 and von Willebrand factor, an endothelial cell marker, were occasionally colocalized in WP bodies. The increase in WP bodies correlated with high ET-1 concentration in the cultured medium, suggesting that these inclusions are involved in storage and release of ET-1 in a manner indicating a regulatory pathway. During the third stage, fibronectin and interleukin (IL)-1alpha were expressed in HUVECs as well as in multinucleated giant cells, which originated from HUVECs, but WP bodies decreased in number in association with a decrease in ET-1 immunoreactivity and concentration. The foregoing changes in immunoreactivities for ET-1, fibronectin, and IL-1alpha affecting HUVECs under prolonged culture may reflect a senescent process of these cells. PMID- 11479773 TI - Ultrastructural identification of iron and copper accumulation in the liver of a male patient with Wilson disease. AB - There is accumulating evidence that ceruloplasmin, a copper-containing protein with ferroxidase activity, plays an important role in iron metabolism. Reduction of ferroxidase activity secondary to ceruloplasmin deficiency may induce iron accumulation in various organs as the result of impaired iron transport. A 37 year-old man presented with intention tremor of the right hand. Liver function tests were almost normal, but parameters of trace elements were abnormal: hypocupremia, hypoceruloplaminemia, and hyperferritinemia. Imaging of the abdomen showed a cirrhotic liver with increased density. A diagnosis of the neurological form of Wilson disease was confirmed by copper deposits in the liver obtained by a blind biopsy, and the patient was diagnosed as compound heterozygous for ATP7B mutations. He was treated with 2500 mg/day trientine hydrochloride per os. The second examination was performed after 20 months of treatment. The treatment further reduced serum ceruloplasmin level from 8.9 to less than 4.0 mg/dl. Serum ferroxidase activity was as low as 70 U/l during treatment. Posttreatment liver histology became negative for copper but remained positive for iron. Copper X rays from hepatocyte lysosomes were no longer detected, but the iron X-ray was still very high post treatment. Thus, microanalysis confirmed compound overload of copper and iron in this male patient with Wilson disease. PMID- 11479774 TI - Electron microscopic and cytochemical studies of peroxidase-negative acute nonlymphoblastic leukemia. AB - Cytochemical and ultrastructural studies of 42 patients with acute nonlymphoblastic leukemia (ANLL) lacking myeloperoxidase as detected by light microscopy were performed. These ANLL patients were classified into minimally differentiated acute myeloid leukemia (AML-M0) (26 cases), mixed-lineage leukemia (9 cases), and acute undifferentiated leukemia (AUL) (7 cases). Ultrastructural morphology revealed blasts from AML-M0 as very immature myeloid cells, blasts from mixed-lineage leukemia as having partially lymphoid cell features, and blasts from AUL as undifferentiated blasts with myeloid nature to some extent. Ultrastructurally, myeloperoxidase activity was positive in 81% of AML-M0 patients and less positive in the mixed-lineage leukemia and AUL patients. The DNA in the nuclei of the AML-M0 blasts was dispersed, as was observed in the immature myeloid cells. Some of the blasts from the mixed-lineage leukemia patients showed condensation of DNA, resembling blasts from acute lymphoblastic leukemia (ALL). The distribution of periodate-reactive glycoconjugates revealed that glycogen particles were absent or, if present, very scanty in AML-M0. Most of the blasts from the AUL patients lacked glycogen particles, and the blasts from those with mixed-lineage leukemia contained clustered glycogen particles, as was observed in the blasts from all patients. These findings disclosed that three subtypes of immature ANLL showed different cytochemical and ultrastructural features in accordance with their immunophenotypical classification. PMID- 11479775 TI - Statistical analysis of endocrine granule size in gastrointestinal carcinoids. AB - Regarding the size of endocrine granules in endocrine neoplasms including carcinoids, most authors have reported only the range of granule diameters, and few studies on detailed morphometry of granule sizes have been documented. The statistically comparative study of endocrine granule sizes among such neoplasms, therefore, presented many difficulties. In the present study, we investigated more detailed findings on the endocrine granule sizes by using statistical analyses of diameters of numerous (n > 200 from each tumor) endocrine granules measured randomly throughout the cytoplasm in 18 gastrointestinal carcinoids. The measured data were analyzed statistically to obtain the histogram, distribution curve, mean value, and standard deviation of granule diameters in each carcinoid. It was disclosed by the analyses that all rectal carcinoids possessed uniformly small endocrine granules in contrast to gastric and duodenal carcinoids, which showed wide distribution of granule sizes, having significantly greater diameters than those in the rectal carcinoid series. The correlation between endocrine granule sizes and producing hormone(s) was, however, not confirmed. PMID- 11479776 TI - Researching basic concepts in clinical homeopathy. PMID- 11479778 TI - Homeopathic pathogenetic trials of Acidum malicum and Acidum ascorbicum. AB - Two homeopathic pathogenetic trials (HPTs, provings), of identical design were conducted: of Acidum malicum 12 cH and Acidum ascorbicum 12 cH. Each trial included 20 healthy volunteers. Both were of double-blind, placebo-controlled, randomised, four period crossover design, with two sequences. 'Healthy' was defined in terms of SF-36 scores, medical judgement and blood tests. All volunteers had regular interviews with the same supervisor. No serious adverse reactions occurred. The causal relationship of each symptom with treatment was judged, blind, by the volunteer, the supervisor and a 9-item pathogenetic index. For Acidum malicum 79 symptoms were identified by the supervisor, 57 were included in the final analysis, 22 occurred in verum treatment periods. For Acidum ascorbicum, of 55 symptoms, 39 were included in the analysis. 16 occurred in verum treatment periods. PMID- 11479779 TI - Can homeopaths detect homeopathic medicines? A pilot study for a randomised, double-blind, placebo controlled investigation of the proving hypothesis. AB - Homeopaths believe that a medicine, which causes a particular symptom in a healthy volunteer, will cure a similar symptom in a sick patient. From this phenomenon, it is possible to deduce a hypothesis: homeopaths should be able to distinguish a homeopathic medicine from a placebo by taking both and observing their effects. If true, this would support an effect of homeopathic medicines different from that of placebo. If false, it casts serious doubt on the contemporary homeopathic knowledge base and on homeopathic pathogenetic trials (HPTs) as currently practised. The study design was a double-blinded, crossover trial. It consisted of a 1-week study medication period, a 2-week washout period and a further 1-week on study medication. Bryonia in a 12c potency was chosen as the trial medication. Participants were recruited in the UK from the Faculty of Homeopathy and the Society of Homeopaths and were currently healthy, aged 18 or over with at least three years' clinical experience of homeopathy. Of the 500 recruitment packs despatched, we received 88 responses (17.6%). Seventy homeopaths were randomised of whom 50 completed the trial. In the main analysis 60% correctly identified the bottle containing Bryonia (n=40; 95% confidence interval 43%, 75%; P=0.27). There was evidence of an order effect: subjects were much more likely to think they received active Bryonia in the first rather than the second period. In this study a promising trend was observed that symptoms reported by some homeopaths may not be completely attributable to placebo. A multi-national, large-scale trial will be required to investigate this phenomena with adequate statistical power. PMID- 11479780 TI - The Constitutional Type Questionnaire: validation in the patient population of the Royal London Homoeopathic Hospital. AB - In homeopathy the choice of a medicine is based on the total 'picture' presented by the patient. This picture includes 'constitutional type' which comprises personality, and general physical features. The Constitutional Type Questionnaire (CTQ) is designed to systematically assess constitutional types. This study examines the reliability and validity of the CTQ. Four hundred and seventy-two outpatients attending clinics at the Royal London Homoeopathic Hospital completed the CTQ, a 152-item scale rating features traditionally considered typical of 19 constitutional homeopathic medicine 'pictures' on 5-point frequency or severity scales. A subsample was retested after 1 week to measure the test-retest reliability. Another subsample was prescribed a medicine by a homeopathic doctor. Prescriptions were compared with the CTQ, to assess the content validity of the scale. The construct validity was measured by Grade of Membership (GOM) analysis. The scale demonstrated good test-retest reliability (r=0.73), internal consistency (r=0.95). The correlation between CTQ results and the medicine prescribed by the homeopathic doctor was 75.8%. The GOM analyses are reported by Davidson et al elsewhere in this issue of the journal. Although the CTQ could be improved, the scale displays good reliability and validity. PMID- 11479781 TI - Do constitutional types really exist? A further study using grade of membership analysis. AB - 'Constitutional types' are a feature of homeopathy. Constitutions are constellations of mental, physical and general features. We present results of a 152-item 'Constitutional Type Questionnaire' (CTQ), relating to 19 major constitutional remedies. Patients referred to the Royal London Homoeopathic Hospital (RLHH) were recruited from outpatient clinics. Each remedy was assigned eight associated features. Each item is rated as it applies to the subject most of the time on a five-point scale. Grade of Membership analysis (GOM) was used for evaluation of remedy profiles. Four hundred and forty-seven individuals provided usable data. Demographic data were collected. A nine pure type solution provided the best fit to the data. Few subjects had exclusive 'membership' of one pure type. There are some difficulties in interpretation; for instance some groups were characterised by the absence of 'constitutional' features. Some pure types are coherent, recognisable types. The results give some support to the homeopathic concept of constitution. CTQ results correlate quite well with the actual prescribing of homeopaths. Further validation with other cohorts is required and with refinement it might become a useful screening method. It cannot simulate complex information processing performed by homeopathic practitioners. PMID- 11479782 TI - Effects of homeopathic treatment in women with premenstrual syndrome: a pilot study. AB - Alternative therapies in general, and homeopathy in particular, lack clear scientific evaluation of efficacy. Controlled clinical trials are urgently needed, especially for conditions that are not helped by conventional methods. The objective of this work was to assess the efficacy of homeopathic treatment in relieving symptoms associated with premenstrual syndrome (PMS). It was a randomised controlled double-blind clinical trial. Two months baseline assessment with post-intervention follow-up for 3 months was conducted at Hadassah Hospital outpatient gynaecology clinic in Jerusalem in Israel 1992-1994. The subjects were 20 women, aged 20-48, suffering from PMS. Homeopathic intervention was chosen individually for each patient, according to a model of symptom clusters. Recruited volunteers with PMS were treated randomly with one oral dose of a homeopathic medication or placebo. The main outcome measure was scores of a daily menstrual distress questionnaire (MDQ) before and after treatment. Psychological tests for suggestibility were used to examine the possible effects of suggestion. Mean MDQ scores fell from 0.44 to 0.13 (P<0.05) with active treatment, and from 0.38 to 0.34 with placebo (NS). (Between group P=0.057). Improvement >30% was observed in 90% of patients receiving active treatment and 37.5% receiving placebo (P=0.048). Homeopathic treatment was found to be effective in alleviating the symptoms of PMS in comparison to placebo. The use of symptom clusters in this trial may offer a novel approach that will facilitate clinical trials in homeopathy. Further research is in progress. PMID- 11479783 TI - Homeopathy in Cuban epidemic neuropathy: an open clinical trial. AB - In an outbreak of epidemic neuropathy (EN) in Cuba (1992-1993), most patients were improved by vitamin therapy. In subjects with residual symptoms, alternative treatments including homeopathy were suggested to ameliorate optic and peripheral signs of the disease. An open clinical pilot trial was conducted on 31 patients with long standing symptoms of optic (OPTI group, n=15) or peripheral EN (PERI group, n=16). During the trial, OPTI and PERI patients continued the same treatment that they received before. Carboneum sulphuratum and Tabacum in homeopathic dilutions were administered for 30 days. These medicines are specific to optic EN, but not closely linked with peripheral EN. Clinical status was evaluated by neurological and ophthalmologic tests at diagnosis (Ddiag), 7 days before homeopathic treatment (D0) and 90 days after (D90). From D0 to D90, the percentages of improvement were 73.3% for the OPTI form and 12.5% for the PERI form. The percentage of improved OPTI patients was significantly higher after the homeopathic treatment vs the period between Ddiag and D0 for optical EN (P<0.01), but not for PERI subjects (P>0.05). In the OPTI group, colour vision, visual acuity and visual field improved after homeopathic treatment (P<0.001), these parameters did not change between Ddiag and D90 (P>0.05). Carboneum sulphuratum and Tabacum showed a reasonable effectiveness in optical EN, but were not effective in PERI EN. PMID- 11479784 TI - Patient benefit survey: Liverpool Regional Department of Homoeopathic Medicine. AB - We report an Outcome Survey carried out at Liverpool Regional Department of Homoeopathic Medicine, over a 12-month period between 1 June 1999 and 31 May 2000, using self-assessment by the Glasgow Homoeopathic Hospital Outcome Score (GHHOS). Overall 76.6% of patients reported an improvement in their conditions since starting homeopathic treatment, while 60.3% scored +2,+3 or+4 on the GHHOS. Fifty-two percent of patients reduced their conventional medication. PMID- 11479785 TI - 20 years ago: the British Homeopathic Journal, July 1981. PMID- 11479789 TI - A proving of Thiosinamine. PMID- 11479791 TI - The research agenda. PMID- 11479792 TI - Definition of homeopathy. PMID- 11479825 TI - [Did deinstitutionalization studies achieve their objective?]. PMID- 11479826 TI - ["It may be somewhat more demanding sometimes, but also more interesting". Psychiatrists evaluate the impact of psychoeducation on outpatient treatment of schizophrenia]. AB - PURPOSE: Aim of the study is the analysis of the psychiatrists' evaluation of the impact of a psychoeducative group programme on patients with schizophrenia and schizoaffective disorders, which was part of the project "Integrated Treatment of Schizophrenia". METHOD: Semi-structured interviews were carried out with 20 psychiatrists involved in outpatient treatment of patients who participated in the psychoeducative programme. Interviews were analysed by means of qualitative content analysis. RESULTS: For some patients psychiatrist noticed positive changes of the awareness for early signs of psychosis, the illness behaviour and their attitudes regarding their disorder. Although only some of the psychiatrists believe that psychoeducation has a direct impact on the reduction of relapse rates and the necessity of inpatient episodes, most of them regard the observed effects as an important improvement of the patients' ability to cope with their disorder. CONCLUSIONS: The effectiveness of psychoeducational programmes should not only be assessed by their impact on the reduction of relapse rates and health service cost but also by its contribution to the patients' quality of life. The integration of psychoeducational programmes into existing outpatient services seems to be an important task of future service planning. PMID- 11479827 TI - [Naltrexone in the treatment of dissociative disorders--reflections with regard to a comprehensive therapeutic concept of borderline disorders]. AB - OBJECTIVE: Not until one year ago has it become more and more known in German speaking countries, that the treatment of borderline and posttraumatic disorders benefits from the use of opiate receptor antagonists. Prerequisite is, that psychotherapeutic approaches, pharmacotherapy and brain physiological research are integrated in an overall concept. METHODS: We report on the current standing of scientific research and moreover our own clinical experiences with opiate antagonists are explained. CONCLUSIONS: Our achieved knowledge permits our to present criteria how to combine these drugs and psychotherapy successfully. PMID- 11479828 TI - [Motor behavior in depression: applications and limitations of actigraphic analyses]. AB - OBJECTIVE: Alterations of motor behavior in depression play a key role in psychosocial activities and interpersonal communication and are relevant for differential diagnosis and treatment. Functions and application of actigraphy and empirical findings in depression will be summarised and discussed. METHODS: The current knowledge retrieved from information of manufacturers, Medline, PsycLIT, and PSYNDEX regarding features of actigraphic equipment and clinical aspects of gross motor analysis in depression was examined. RESULTS: Actigraphy is a practical method to monitor and quantify gross motor activity, but setting variables including sensitivity or epoch length vary among commercially available instruments. Measures of motor activity in depressed patients are used for diagnosis, evaluation of sleep, and prediction of treatment response. CONCLUSIONS: Measurements of motor behavior by actigraphy can be used in depression for diagnostic procedures and treatment evaluation. The role of actigraphic setting variables needs further clarification. Actigraphy may be used for behavioral analysis including individual regulation of motor activity as an objective parameter for evaluation of depression and other psychiatric disorders. PMID- 11479829 TI - [Number of hospitalizations according to German "PsychKG" legislation has doubled in 10 years]. AB - AIM: to examine the frequency of involuntary hospitalisation according to PsychKG (german law concerning psychiatric practice) in a comprehensive region with good overall medical/psychiatric services, over the course of a decade. METHOD: All cases of compulsory admission into Psychiatric Clinics within the greater Gottingen area (Central Germany) were evaluated every two years between 1988 and 1998. RESULTS: The number of involuntarily hospitalised patients doubled within the ten years of evaluation although the density of population remained stable and the already good quality out-patient treatment experienced further improvement. However, the total number of in-patients also doubled during this time, so that the proportion between compulsory admission and voluntary hospitalisation remained the same. CONCLUSION: Similar to other regions, the increase in compulsory admissions is considerable. The development towards shorter treatments and an increasing frequency of re-hospitalisation seem to have no significant influence on the courses of disease or on future pathological crises of the patients in question. It is possible that the expected lack of a decreasing rate of compulsory admission is partly caused by behaviour on the part of the physician. PMID- 11479830 TI - [Heinrich von Kleist--crisis and creative overcoming]. AB - Heinrich von Kleist's life was shaken repeatedly by negative life-events, finally he committed suicide in his last life-crisis (1811). His work was mostly understood as descriptions of negative life-events and failed-being. In this article it will be shown that in at least two "crises" Kleist's work can be understood as a creative overcome of those. Kleist shows in his "Essay to Find a Sure Way to Happiness" (1799) his way of solving his "Soldier-Crisis" (1798), a depressive episode. In "The Broken Jug" (1802 - 1805) he shows the implications of a philosophical problem experienced in his "Kant-Crisis" (1801) and offers chances to overcome this particular crisis, which still seems to be an actual problem of ourselves. Though his crises must be understood as depressive episodes, at least the "Kant-Crisis" with its connections to philosophical and artistical matters seems to be more complicated than a simple depressive syndrome. Kleist formulates his basic life-experience, to be repeatedly shaken by "crises" respectively depressive episodes and the necessity to overcome each in a new way of living. PMID- 11479831 TI - [Long-term patients in supported housing after deinstitutionalisation--part V of the Berlin Deinstitutionalisation Study]. AB - OBJECTIVE: In part IV of the Berlin Deinstitutionalisation Study, we reported a positive change in quality of life for 65 resettled patients one year after discharge. In this part, we investigated changes in a longer follow-up period. METHODS: A group of 66 patients discharged into supported housing was assessed one and five years after baseline. Quality of life, needs for care, patients' assessment of treatment, re-hospitalisation and characteristics of care were studied in a control-group design. RESULTS: After five years, 61 % of the patients continued to live in highly staffed settings (mean staff-patient-ratio: 1 : 3.5). Eleven percent had moved on to independent living and 16 % had returned into an institutional setting. Standard mortality ratios were 4.4 (all causes of death) and 62.5 (2 suicides only of schizophrenic patients). After one year patients (n = 20) showed a significant increase in satisfaction with their living situation and more social contacts. Over the longer period of time, there were also favourable changes, which failed to reach statistical significance (n = 24). Average length of inpatient stay per year of the illness was decreased significantly at both follow-ups. CONCLUSIONS: Interpretation of the findings is difficult because of the small sample size and selection effects. Most of the former long-stay patients appear able to live in the community with a significant decrease of time spent in hospital. Whilst patients were particularly satisfied with their accommodation at one year follow-up, this did not lead to a generalised improvement in subjective quality of life and other subjective criteria in the long term. In line with other recent studies on community mental health care one may conclude that small case loads alone do not have major beneficial effects. Further research should evaluate more systematic and targeted models of mental health care in the community in general and of supported housing in particular. PMID- 11479832 TI - [Treatment of panic disorder with combination of SSRI and cognitive-behavioral therapy]. AB - GOAL: Various forms of treatment of panic disorder with and without agoraphobia have shown good results. We examined the combination of cognitive-behavioral psychotherapy and treatment with an SSRI. METHODS: Case report and literature review. RESULT: The combination treatment shows a faster onset of therapeutic effects than cognitive-behavioral psychotherapy. However, final outcomes are comparable. CONCLUSIONS: The addition of a SSRI to cognitive-behavioral psychotherapy accelerates the onset of therapeutic effects. It does not interfere with the cognitive restructuring aimed at cognitive-behavioral psychotherapy. PMID- 11479833 TI - [Aphasia and dementia]. AB - Primary progressive aphasia (PPA) is an uncommon neurodegenerative syndrome characterized by a relatively isolated dissolution of language function at the beginning, followed by deterioration of general cognitive function and of activities of daily living after 2 or more years. On account of neuropathological and clinical findings, PPA is supposed to form part of the spectrum of frontotemporal lobar degeneration. We present a case study of a 66-year-old woman with a probable fluent progressive aphasia. She initially experienced word amnesia and developed after 2 - 3 years gradual regression of word comprehension, over-fluent speech with semantic paraphasias, and at last generalized dementia. In addition to minor bilateral cortical volume reduction on CCT, MRI showed left temporal lobe atrophy involving hippocampus, SPECT revealed reduced uptake left frontal and temporal. PMID- 11479834 TI - Quantitative similarity-based association tests using population samples. AB - Although genetic association studies using unrelated individuals may be subject to bias caused by population stratification, alternative methods that are robust to population stratification, such as family-based association designs, may be less powerful. Furthermore, it is often more feasible and less expensive to collect unrelated individuals. Recently, several statistical methods have been proposed for case-control association tests in a structured population; these methods may be robust to population stratification. In the present study, we propose a quantitative similarity-based association test (QSAT) to identify association between a candidate marker and a quantitative trait of interest, through use of unrelated individuals. For the QSAT, we first determine whether two individuals are from the same subpopulation or from different subpopulations, using genotype data at a set of independent markers. We then perform an association test between the candidate marker and the quantitative trait, through incorporation of such information. Simulation results based on either coalescent models or empirical population genetics data show that the QSAT has a correct type I error rate in the presence of population stratification and that the power of the QSAT is higher than that of family-based association designs. PMID- 11479835 TI - Hereditary sensory neuropathy type I: haplotype analysis shows founders in southern England and Europe. AB - Hereditary sensory neuropathy type I (HSN1) is the most common dominantly inherited degenerative disorder of sensory neurons. The gene mutation was mapped to chromosome 9 in a large Australian family, descended from an ancestor from southern England who was a convict. Dawkins et al. recently reported gene mutations in the SPTLC1 gene, in this and other families. The first description of hereditary sensory neuropathy, by Hicks, was in a family from London and Exeter. To determine if the families in the present study that have SPTLC1 mutations are related to English families with HSN1 and, possibly, to the family studied by Hicks, we performed haplotype analysis of four Australian families of English extraction, four English families, and one Austrian family. Three Australian families of English extraction and three English families (two of whom have been described elsewhere) had the 399T-->G SPTLC1 mutation, the same chromosome 9 haplotype, and the same phenotype. The Australian and English families may therefore have a common founder who, on the basis of historical information, has been determined to have lived in southern England prior to 1800. The sensorimotor neuropathy phenotype caused by the 399T-->G SPTLC1 mutation is the same as that reported by Campbell and Hoffman and, possibly, the same as that originally described by Hicks. PMID- 11479836 TI - Optimized group sequential study designs for tests of genetic linkage and association in complex diseases. AB - The study of genetic linkage or association in complex traits requires large sample sizes, as the expected effect sizes are small and extremely low significance levels need to be adopted. One possible way to reduce the numbers of phenotypings and genotypings is the use of a sequential study design. Here, average sample sizes are decreased by conducting interim analyses with the possibility to stop the investigation early if the result is significant. We applied optimized group sequential study designs to the analysis of genetic linkage (one-sided mean test) and association (two-sided transmission/disequilibrium test). For designs with two and three stages at overall significance levels of.05 and.0001 and a power of.8, we calculated necessary sample sizes, time points, and critical boundaries for interim and final analyses. Monte Carlo simulation analyses were performed to confirm the validity of the asymptotic approximation. Furthermore, we calculated average sample sizes required under the null and alternative hypotheses in the different study designs. It was shown that the application of a group sequential design led to a maximal increase in sample size of 8% under the null hypothesis, compared with the fixed-sample design. This was contrasted by savings of up to 20% in average sample sizes under the alternative hypothesis, depending on the applied design. These savings affect the amounts of genotyping and phenotyping required for a study and therefore lead to a significant decrease in cost and time. PMID- 11479837 TI - Cervical spine injuries in children: a review of 103 patients treated consecutively at a level 1 pediatric trauma center. AB - PURPOSE: Cervical spine (C-spine) injuries occur infrequently in children but may be associated with significant disability and mortality. The purpose of this study was to review the experience of a level 1 pediatric trauma center to determine the epidemiology, risk factors, mechanisms, levels, types of injury, comorbid factors, and outcomes associated with these potentially devastating injuries. METHODS: A retrospective analysis of 103 consecutive C-spine injuries treated at a level 1 pediatric trauma center over a 9(1/2)-year period (January 1991 through August 2000) was performed. RESULTS: The mean age was 10.3 +/- 5.2 years, and the male-to-female ratio was 1.6:1. The most common mechanism of injury was motor vehicle related (52%), followed by sporting injuries (27%). Football injuries accounted for 29% of all sports-related injuries. Sixty-eight percent of all children sustained injuries to C1 to C4; 25% to C5 to C7; and 7% to both. Spinal cord injury without radiographic abnormality (SCIWORA) occurred in 38%. Five patients had complete cord lesions involving the lower C-spine (C4 to C7); 4 of these were motor vehicle related, and all 4 patients died. Isolated C-spine injuries occurred in 43%, whereas 38% had associated closed head injuries (CHI). The overall mortality rate was 18.5%, most commonly motor vehicle related (95%), occurring in younger children (mean and median age 5 years) and associated with upper C-spine injuries (74%) and CHI (89%). C1 dislocations occurred in younger children (mean age, 6.6 years), most often as a result of motor vehicle related trauma (especially pedestrians) and were associated with the highest injury severity score (ISS), longest length of stay (LOS), most CHIs, and the highest mortality rate (50%). C-spine fractures with or without SCI occurred most commonly as a result of falls and dives. Sporting injuries occurred almost exclusively in adolescent boys (mean age, 13.8 years) and were isolated injuries associated with a relatively low ISS and shorter LOS. Interestingly, 75% of sporting injuries showed SCIWORA, and all infants suffering from child abuse had SCIWORA. CONCLUSIONS: Mechanisms of injury are age related, with younger children sustaining C-spine injuries as a result of motor vehicle-related trauma and older adolescents commonly injured during sporting activities. C-spine injuries in children most commonly involve the upper C-spine, but complete lesions of the cord are associated more frequently with lower C-spine injuries. The type of C spine injury is related to the mechanism of injury: SCIWORA is associated with sporting activities and child abuse, C-spine dislocations most commonly result from motor vehicle-related trauma (especially among pedestrians), and C-spine fractures occur most commonly as a result of falls and dives. Predictors of mortality include younger age, motor vehicle-related mechanism, C1 dislocations, high ISS greater than 25, and associated CHI. A high index of suspicion for SCIWORA is essential when evaluating adolescents with neck trauma associated with sporting injuries or victims of child abuse. PMID- 11479838 TI - Life-threatening dog attacks: a devastating combination of penetrating and blunt injuries. AB - BACKGROUND/PURPOSE: Children often are the victims of dog attacks. Although bite injuries sustained in an attack characteristically are attributed to the penetrating component of the bite, the blunt nature of a bite may represent the most serious and devastating component of injury. The purpose of this study was to characterize a group of children suffering life-threatening dog bites and examine the predominant aspect of injury. METHODS: Thirty-nine children were admitted to the trauma service at a regional pediatric trauma center with the diagnosis of dog bite injury over a 6-year period (1994 through 1999). Patient demographics, site and description of injury, and surgical procedures performed were recorded from a chart review. RESULTS: Mean age of the 35 children included for analysis was 5.4 years (range, 0.8 to 17 years). Twenty-five (71%) injuries occurred in the head and neck region. Eight (23%) children sustained life threatening injuries. Of these, blunt force was the predominant injury in 6. This resulted in 1 (20%) arterial occlusion requiring vascular reconstruction, 2 (40%) permanent neurologic injuries (stroke, spinal cord transection), and 1 (20%) death (exsanguination). CONCLUSIONS: On evaluation of a dog attack, the focus generally is on the obvious penetrating aspect of the bite. Yet, we found the blunt component of injury can have devastating consequences reflected in acute arterial, brain, and spinal cord injury. Even in the absence of significant penetrating trauma, further evaluation should be considered to exclude occult blunt arterial or neurologic injury. PMID- 11479839 TI - The use of a bioactive skin substitute decreases length of stay for pediatric burn patients. AB - BACKGROUND: To optimize burn care for children, the authors introduced a protocol incorporating the use of a bioactive skin substitute, TransCyte (Advanced Tissue Sciences, La Jolla, CA). This study was designed to determine whether this management plan was safe, efficacious, and decreased hospital inpatient length of stay (LOS) compared with conventional burn management in children. METHODS: All pediatric burns greater than 7% total body surface area (TBSA) that occurred after October 1999 underwent wound closure with TransCyte (n = 20). These cases were compared with the previous 20 consecutive burn cases greater than 7% TBSA that received standard therapy. Standard therapy consisted of application of antimicrobial ointments and hydrodebridement. The following information was obtained: burn mechanism, age, size of burn, requirement of autograft, and LOS. Data were analyzed using the student's t test. RESULTS: Data for age, percent TBSA burn and LOS are reported as means +/- SEM. The children who received standard therapy were 2.99 +/- 0.7 years compared with those receiving TransCyte were 3.1 +/- 0.8 years. There was no difference between the treatment groups with regard to percent TBSA burn: standard therapy, 14.3 +/- 1.4% TBSA versus TransCyte, 12.7 +/- 1.3% TBSA. There was no difference in the type of burns in each group, the majority were liquid scald type, 70% in the standard therapy group versus 90% in the TransCyte group. Only 1 child in the TransCyte group required autografting (5%) compared with 7 children in the standard therapy group (35%). Children treated with TransCyte had a statistically 6 significant decreaed LOS compared with those receiving standard therapy, 5.9 +/- 0.9 days versus 13.8 +/- 2.2 days, respectively (P =.002). CONCLUSIONS: This is the first study using TransCyte in children. The authors found that this protocol of burn care was safe, effective, and significantly reduced the LOS. This new approach to pediatric burn care is effective and improves the quality of care for children with burns. PMID- 11479840 TI - Intestinal cytokine gene expression in infants with acute necrotizing enterocolitis: interleukin-11 mRNA expression inversely correlates with extent of disease. AB - BACKGROUND/PURPOSE: The authors have shown previously that surgical specimens from infants with acute necrotizing enterocolitis (NEC) show upregulation of inducible nitric oxide (NO) synthase (iNOS) and interferon-gamma mRNA. However, the contribution of other inflammatory cytokines such as interleukin-8 (IL-8), IL 11, and IL-12 has not been defined. Likewise, the role of GTP-cyclohydrolase, the rate-limiting enzyme in tetrahydrobiopterin synthesis, and thus NO production by iNOS is unclear. In this study, the authors sought to further define the pattern of cytokine expression seen in infants with acute NEC. METHODS: The authors measured intestinal cytokine mRNA expression by semiquantitative reverse transcriptase polymerase chain reaction in 21 infants with histologically confirmed NEC, 18 with other inflammatory conditions, and in 9 patients without intestinal inflammation. Guanosine triphosphate-cyclohydrolase (GTP-CH) activity was measured by specific enzyme assay. Univariate exact logistic regression analysis was performed to identify predictors of outcome. RESULTS: IL-8 and IL-11 mRNA were upregulated in patients with acute NEC compared with those with other inflammatory conditions or those without disease; these levels returned to baseline at the time of stoma closure. Increased IL-11 mRNA decreased the likelihood of pan-necrosis (odds ratio, 0.93; P =.002). Increased IL-12 levels (but not IL-8) seemed to protect against pan-necrosis (odds ratio, 0.70; P =.06). CONCLUSIONS: Local upregulation of IL-11 may represent an adaptive response designed to limit the extent of intestinal damage in NEC. Decreased IL-12 levels may contribute to the pathogenesis of NEC by allowing bacteria to escape host defenses. PMID- 11479841 TI - Heparin-binding EGF-like growth factor decreases apoptosis in intestinal epithelial cells in vitro. AB - BACKGROUND/PURPOSE: The production of heparin-binding EGF-like growth factor (HB EGF) is upregulated during organ injury and has a cytoprotective effect during hypoxic stress in intestinal epithelial cells in vitro and intestinal ischemia reperfusion injuries in vivo. The purpose of this study was to determine if HB EGF-related cytoprotection is manifested through alterations in apoptosis. METHODS: Human intestinal epithelial cell monolayers (DLD-1 and Caco-2) were stimulated with interleukin (IL)-1 (20 ng/mL), tumor necrosis factor (TNF)-alpha (40 ng/mL), and interferon (IFN)-gamma (10 ng/mL) with or without HB-EGF (1, 10 or 100 ng/mL) and analyzed for rates of apoptosis utilizing a Cell Death Detection ELISA and flow cytometry. RESULTS: ELISA results showed a 3-fold increase in the level of apoptosis during stimulation with cytokines compared with nonstimulated cells (P <.05). Relative levels of cytokine induced apoptosis were reduced after 12 hours of HB-EGF exposure (P <.05) in a dose-dependent fashion. Results of flow cytometric analysis also showed a reduction in apoptosis at 6 hours when cell monolayers were stimulated with cytokines in conjunction with HB-EGF compared with cytokines alone (P <.05). CONCLUSIONS: HB-EGF downregulated apoptosis in intestinal epithelial cells exposed to proinflammatory cytokines in vitro. The results of this study suggest that alterations in apoptosis may represent a possible mechanism by which this growth factor exerts its cytoprotective effect at the mucosal level during the proinflammatory state. PMID- 11479842 TI - Attenuated expression of inducible nitric oxide synthase in lung microvascular endothelial cells is associated with an increase in ICAM-1 expression. AB - BACKGROUND/PURPOSE: The molecular and cellular events that regulate inflammatory lung injury, a major cause of morbidity in surgical patients, remain unclear. The authors hypothesize that nitric oxide (NO) plays an important role in regulating polymorphonuclear cell (PMN)-induced acute lung injury, and further, that attenuated expression of inducible nitric oxide synthase (iNOS) and therefore decreased production of NO by lung microvascular endothelial cells (LMVEC), accelerates inflammation and injury. METHODS: LMVEC and aortic EC (AEC) from rat and human were stimulated with lipopolysaccharide (LPS) and cytokines; changes in iNOS mRNA expression and iNOS activity were determined. The role of NO in mediating inflammatory responses was evaluated by determining PMN adherence to LMVEC and lung tissue slices in the presence and absence of NOS inhibitors and NO donors. Human LMVEC and AEC were assessed by FACS analysis for ICAM-1 expression, because this is thought to be a critical determinant of PMN adherence. RESULTS: When stimulated with endotoxin and cytokines, rat AEC monolayers express nearly 3 fold more iNOS mRNA than rat LMVEC. The low levels of LMVEC iNOS expression are associated with a 4-fold lower nitrite and nitrate production. Similar trends are seen in human endothelial cells. When iNOS activity was blocked, PMN adherence to tumor necrosis factor alpha (TNFalpha)/LPS-stimulated LMVEC was markedly increased. In contrast, adding a nitric oxide donor to endotoxin/cytokine stimulated LMVEC monolayers reduced PMN adherence to near background levels. Similar responses were observed in vivo. Human lung microvascular endothelial cells show a substantially increased level of ICAM-1 upregulation when compared with similarly stimulated human aortic macrovascular endothelial cells. CONCLUSIONS: These data indicate that LMVEC express less iNOS and produce less NO than AEC. This lower expression and activity of iNOS in LMVEC may be linked to increased expression of ICAM-1. Because ICAM-1 has been shown to be essential for tight PMN adherence, these data suggest that relatively low iNOS expression in LMVEC may contribute to a propensity for the lung to be injured by activated PMNs. PMID- 11479843 TI - Portal venous air: the poor prognosis persists. AB - BACKGROUND/PURPOSE: The prognostic importance of portal vein air (PVA) in babies with necrotizing enterocolitis (NEC) has been controversial. This study compares the outcome in babies with NEC and PVA treated surgically versus those with medical management. METHODS: Forty neonates in the neonatal intensive care unit (NICU; 1995 through 1999) had (PVA) during their hospitalization. Babies were analyzed for gestational age (GA), birth weight (BW), and survival after operative versus medical management. RESULTS: The average GA was 26 weeks, average BW was 1,173 g. Twenty-three patients (57.5%) tolerated full feedings and 8 (20%) partial feedings at diagnosis. All 40 babies required intubation at birth with 23 (57.5%) requiring reintubation with onset of PVA. In all cases, PVA was present within 24 hours of onset of abdominal distension, feeding intolerance, or heme-positive stools. Two cases of PVA "resolved" only to recur later in the patients' courses. Thirty-two patients (80%) manifested pneumatosis intestinalis on abdominal radiographs, and 8 (20%) had perforations. Acidosis was present in 25 (63%) patients, and vasopressor support (dopamine) was required in 15 (38%), with 2 patients requiring support only preoperatively. Initial management consisted of bowel rest, fluid resuscitation, orogastric decompression, and broad spectrum antibiotics. Operation was performed in 31 (78%). Seventeen underwent resection with ostomy formation with 6 deaths and 11 survivors. Four underwent resection using the clip and drop back method, with one death and 3 requiring an ostomy at second look laparotomy. Ten had NEC totalis and closure of the abdomen only. Overall operative mortality rate was 17 of 31 (54%). Nine seemingly stable patients were treated nonoperatively. Six had progressed disease and died before salvage laparotomy could be performed, whereas 3 (33%) survived without further therapy. CONCLUSIONS: PVA has been a relative indication for operation. This view has been challenged by the survival of some patients without laparotomy. Although nonoperative therapy seems appealing in hemodynamically stable patients without acidosis, our data confirm the poor prognosis of infants with PVA and NEC. PMID- 11479844 TI - Inherited thrombophilia: a possible cause of in utero vascular thrombosis in children with intestinal atresia. AB - BACKGROUND: Congenital atresia of the small and large intestine is thought to evolve from in utero mesenteric vascular occlusion of the corresponding intestinal segment. Because spontaneous thrombosis recently has been described in association with inherited thrombophilia, the authors wondered if inherited thrombophilia also might be found in babies with intestinal atresia. METHODS: Genetic analysis was done on 28 children treated for congenital intestinal atresia. DNA was analyzed for point mutations to detect the 2 most common types of inherited thrombophilia, the G1691A mutation in the factor V gene (factor V Leiden) and the G20210A mutation in the prothrombin gene. In addition, other genetic risk factors for thrombosis were analyzed including the C677T mutation in the methylenetetrahydrofolate reductase gene (MTHFR) and 2 polymorphisms of the factor VII gene (the R353Q and the hypervariable region 4 polymorphisms). RESULTS: The factor V Leiden mutation was present in 5 of 28 (18%) children treated for congenital intestinal atresia. This is increased significantly when compared with the reported carrier frequency of 3% to 7% in the general population and a reported carrier rate of 4.2% in the local population (P <.005). The R353Q polymorphism of the factor VII gene, specifically the RR genotype, was noted in 85% of patients with atresia with an expected frequency of 64% (P <.008). There were no significant associations noted between mutations in the prothrombin gene, the MTHFR gene, or the hypervariable region of the factor VII gene. CONCLUSIONS: The factor V Leiden mutation and the RR subtype of the R353Q polymorphism of the factor VII gene are seen at an increased frequency in children with congenital intestinal atresia. This suggests that inherited thrombophilia may play a role in the etiology of these in utero mesenteric thrombotic events. PMID- 11479845 TI - Retinoid signaling directs secondary lineage selection in pancreatic organogenesis. AB - BACKGROUND/PURPOSE: Retinoid signaling plays an important role in many differentiation pathways. Retinoid signaling has been implicated in the induction of differentiation by pancreatic ductal cancer cell lines and in patients with pancreatic cancer. The authors wished to better understand the role of retinoid signaling in pancreatic development. METHODS: Embryonic pancreas was harvested from mice at serial gestational ages and immunohistochemical analysis was performed for retinoic acid receptors (RAR-alpha, RAR-beta, RAR-gamma), and retinoid X receptors (RXR-alpha, RXR-beta, and RXR-gamma). Also, early embryonic pancreases were cultured for 7 days with exogenous 9-cis retinoic acid (9cRA) or all-trans retinoic acid (atRA) and analyzed histologically and immunohistochemically. RESULTS: Retinoid receptors were expressed in a lineage specific distribution, with stronger expression for many in the exocrine compartment. The receptors were not often expressed until late gestation. Exogenous 9cRA induced predominantly ducts instead of acini, plus more mature endocrine (islet) architecture. Exogenous atRA induced predominantly acini instead of ducts, with no apparent endocrine effect. CONCLUSIONS: Retinoids may have an important role in pancreatic differentiation, with a particular effect on secondary lineage selection between ductal and acinar phenotype. Because the control of ductal versus acinar differentiation has been implicated strongly in the pathogenesis of pancreatic ductal carcinoma, these results may lay the groundwork for studies in the mechanism of induced differentiation of pancreatic ductal cancer by retinoids. PMID- 11479846 TI - Preliminary report of electrogastrography in pediatric gastroresection: can it be predictive of alteration of gastric motility? AB - BACKGROUND/PURPOSE: Gastric resection is an infrequent surgical procedure in childhood. However, the use of the stomach for bladder augmentation and substitution is well documented. Partial gastrectomy performed in gastrocystoplasty (GCP) involves the greater curvature of the stomach, the same area in which gastric pace-maker cells are known to be placed. The aim of this study was to assess, by electrogastrography (EGG), if subtotal gastric resection can alter gastric motility in children submitted to partial gastrectomy for GCP. METHODS: Gastric electrical activity (GEA) was evaluated in 25 children using EGG: 10 patients (4 boys, 6 girls; mean age, 11.6 years) previously submitted to GCP, and 15 normal subjects (12 boys, 3 girls; mean age, 8.62 +/- 2.77 years) as controls. All patients were submitted to cutaneous EGG; recording GEA for 30 minutes before and after a standard test meal. The percentage of 3 cycles per minute (3CPM), bradygastria, tachygastria, DFIC (dominant frequency instability coefficient), DPIC (dominant power instability coefficient), PDP (period dominant power), PDF (period dominant frequency) were recorded and analyzed using Wilcoxon matched-pair test. Data were considered statistically significant if P <.05. RESULTS: Normal subjects as well as operated patients showed a statistically significant difference in bradygastria (P =.05), PDP and PDF (P =.05) percentage, comparing pre versus postprandial period. In the normal group, 3CPM (P =.0012) and DFIC (P =.0008) were statistically different between the pre- and postprandial period. Patients who underwent GCP did not show any statistically significant difference in 3CPM and DFIC pre- and postprandial. CONCLUSIONS: In normal subjects, GEA showed a complete variation after the meal, whereas in operated patients GEA was impaired and only partially modified after the meal. This observation suggests that in patients with gastric resection, adaptation of the stomach to food ingestion is present but incomplete with respect to normal subjects; it can be caused by surgical removal of the pace-maker cells of the greater curvature. For this reason a follow-up analysis of gastric function is recommended for all patients undergoing GCP. PMID- 11479847 TI - Methodology standards associated with quality reporting in clinical studies in pediatric surgery journals. AB - BACKGROUND/PURPOSE: Reports of clinical trials often lack adequate descriptions of design and analysis; recent attention has focused on improving this omission so readers can properly assess the strength of the findings and draw their own conclusions. Similar analysis of study design and methodologic standards associated with quality reporting has not been carried out for pediatric surgery journals. METHODS: All studies (n = 642) published in 1998 in Journal of Pediatric Surgery (JPS) and Pediatric Surgery International (PSI), were reviewed for demographic data and study design. The frequency of reporting of 11 basic elements of design and analysis was evaluated in randomized clinical trials (RCT), nonrandomized clinical trials (NRCT), and retrospective cohorts (RC) from JPS by consensus of 2 assessors. RESULTS: Of the 642 studies, 17% of articles (111 of 642) were classified as clinical studies. Sixty-three were comparative studies and consisted of RC (n = 48), NRCT (n = 12), and RCT (n = 3). Two-thirds of articles published were either case reports or case series (431 of 642), and 16% were basic science articles. Demographic analysis showed a wide range of topics addressed, 4 authors per article, and multiple country of origin of authors. More than 66% of all RCT in JPS reported on eligibility criteria, admission before allocation, random allocation, method of randomization, patients' blindness to treatment, treatment complications, statistical analyses, statistical methods, loss to follow-up, and statistical methods; 2 elements of design and analysis, however, were poorly reported: blind assessment of outcome (33%) and power (17%). CONCLUSIONS: There were few randomized, controlled trials in pediatric surgery journals, and further attention should be given to evaluate the causal factors. Nine elements of quality reporting were well reported; however, 2 others were poorly reported; this may improve if editors of pediatric surgical journals provide authors with guidelines on how to report clinical trial design and analysis. PMID- 11479848 TI - Thoracoscopic resection of mediastinal masses in infants and children: an evaluation of technique and results. AB - BACKGROUND: Thoracoscopy has been accepted as a technique in pediatric surgery for diagnosis of thoracic pathology, but there has been little experience using it as a therapeutic modality as well. The purpose of this report is to describe and critically evaluate a 7-year experience with thoracoscopic diagnosis and resection of mediastinal masses in infants and children. METHODS: From February 1993 to June 2000, 39 patients presented with mediastinal masses and no tissue diagnosis. Age ranged from 5 months to 18 years old and weight from 3.6 to 110 kg. Twelve children had anterior mediastinal masses, 27 posterior. The patients were positioned in a modified prone or supine position, and single lung ventilation was performed on the contralateral side. Three or 4 valved trocars were utilized with 3 and 5 mm instrumentation. RESULTS: A total of 38 of 39 procedures were completed successfully endoscopically. The procedure in 1 patient with a sarcoma was converted to thoracotomy because of extensive disease. Operating times ranged from 20 to 185 minutes. Diagnosis was obtained in all cases, and complete excision was performed in 33. All children were extubated in the operating room; 8 chest tubes were placed but removed within 24 hours. Hospital stay ranged from 12 hours to 4 days. Pathology included 12 foregut duplications, 7 ganglioneuromas, 6 neuroblastomas, 6 lymphomas, 3 teratomas, 2 sarcomas, and 3 other lesions. CONCLUSION: Thoracoscopy is a safe and effective method to evaluate, biopsy, and in most cases resect lesions of the anterior and posterior mediastinum in infants and children. PMID- 11479849 TI - Advancement flap umbilicoplasty after abdominal wall closure: postoperative results compared with normal umbilical anatomy. AB - BACKGROUND/PURPOSE: Umbilical reconstruction frequently is an adjunct used after repair of congenital abdominal wall defects. The authors describe a new technique of umbilicoplasty and compare their results with normal neonatal umbilical anatomy. METHODS: After fascial closure, 6 newborn infants with either gastroschisis (n = 3) or omphalocele (n = 3) underwent umbilicoplasty performed by advancing bilateral skin flaps including creation of an umbilical skin collar. The umbilical collar height, circumference, and distance from the xiphoid with respect to the pubis (ratio of xiphoid-umbilicus distance to xiphoid-pubis distance or XU:XP) were recorded and referenced to normal umbilical anatomy in additional 28 infants (24 to 40 weeks gestation [mean, 33.2 weeks] and age 2 to 131 days [mean, 26.5 days]). RESULTS: After umbilicoplasty, all infants had a near-normal-appearing umbilicus (collar height, 0.75 +/- 0.25 cm and circumference, 5.24 +/- 1.20 cm) compared with normal umbilical anatomy (collar height, 4.36 +/- 1.32 cm and circumference, 1.03 +/- 0.38 cm). However, after umbilicoplasty, the neoumbilicus was positioned more cephalad (XU:XP = 0.53 +/ .05) compared with normal (mean XU:XP, 0.67 +/- 0.07). The only complication encountered was a small dehiscence of the superior aspect of the abdominal wound. CONCLUSIONS: Closure of abdominal wall defects with simultaneous umbilicoplasty provides a cosmetically pleasing result. When performing umbilicoplasty, attempt should be made to position the neoumbilicus as inferior as possible, optimally at two thirds the distance from the xiphoid to the pubis. PMID- 11479850 TI - Nutritional morbidity in survivors of congenital diaphragmatic hernia. AB - PURPOSE: The purpose of this report is to detail the nutritional sequelae seen in survivors of congenital diaphragmatic hernia (CDH) followed in a multidisciplinary clinic. METHODS: Data on 121 surviving CDH patients seen between 1990 and 2000 were collected. Regression analysis was used to determine the impact of factors such as Apgar score, birth weight, extracorporeal membrane oxygenation (ECMO), and patch repair on outcomes associated with nutritional morbidity. RESULTS: There were 100 left and 21 right CDH defects. Mean birth weight and 5-minute Apgar score were 3.1 kg (+/-0.8) and 6.8(+/-2), respectively. Extracorporeal membrane oxygenation was required in 43 (36%) patients and patch repair in 39 (32%). A gastrostomy was required in 39 (32%) patients and a fundoplication in 23 (19%) patients. The side of the defect did not affect the frequency of these procedures. Fifty-six percent of patients were below the 25th percentile for weight during most of their first year. Regression analysis found that duration of ventilation (P <.001) and the presence of a patch repair (P =.03) were independent variables predictive of failure to thrive thereby requiring a gastrostomy tube. Patch repair also was predictive of need for subsequent fundoplication caused by gastroesophageal reflux (P <.001). Twenty nine patients (24%) had severe oral aversion. Risk factors were prolonged ventilation (P =.001) and oxygen requirement at discharge (P =.015). Two thirds of these patients subsequently improved. CONCLUSIONS: Nutritional problems continue to be a source of morbidity for survivors of CDH, particularly in the first year of life. Not surprisingly, patients who had prolonged intubation and prosthetic material at the gastroesophageal junction fared worse. Despite aggressive nutritional management, 56% of the population remained below the twenty-fifth percentile for weight. These data show the need for careful nutritional assessment in all CDH patients, especially those at high risk for malnutrition. PMID- 11479851 TI - Combination antiangiogenic therapy: increased efficacy in a murine model of Wilms tumor. AB - BACKGROUND/PURPOSE: Antibody to vascular endothelial growth factor (anti-VEGF) suppresses tumor growth and metastasis in experimental Wilms tumor. However, tumor growth accelerates if antibody is withdrawn. As recently shown, low-dose, frequently administered topotecan, a topoisomerase-1 inhibitor, has anti angiogenic activity. The authors hypothesized that combined topotecan/anti-VEGF therapy would suppress tumor growth and metastasis more durably than either agent alone. METHODS: Xenografts were induced by intrarenal injection of human Wilms tumor cells in athymic mice (n = 59). Mice were divided into control (n = 10), anti-VEGF (n = 16), topotecan (n = 17), and topotecan plus anti-VEGF (n = 16) groups. All control and half the treated mice were killed at week 6. Remaining ("rebound") mice were maintained without treatment until week 8. Tumor vasculature was mapped by fluorescein angiography/PECAM immunostaining. Endothelial apoptosis was assessed by TUNEL assay. RESULTS: 6 weeks: Tumor weights were reduced significantly in treated mice (P <.003 v control). Seven of ten control and 1 of 25 treated mice displayed lung metastases (P <.003). Rebound tumors were largest in topotecan-only, intermediate in antibody-treated, and smallest in combination-treated mice. Immunostaining and angiography results showed sparse vascularity in treated xenografts. Endothelial apoptosis was observed only in treated tumors. CONCLUSION: Combination low-dose topotecan and anti-VEGF antibody therapy is antiangiogenic and suppresses tumor growth and metastasis in experimental Wilms tumor more durably than either agent alone. PMID- 11479852 TI - The role of prospective randomized clinical trials in pediatric surgery: state of the art? AB - PURPOSE: This study sought to determine the role of randomized controlled trials (RCT) in the evolution of pediatric surgical practice. METHODS: The authors used a computer-assisted literature search to identify all clinical trials related to pediatric surgery published in the English-language literature from 1966 through 1999. Each article was reviewed in detail for purpose, content, conduct, and quality of the trial. The authors assessed quality with a previously validated instrument (Chalmers Qualitative Assessment). RESULTS: The authors identified 134 RCTs related to pediatric surgery over the past 33 years. This accounts for 0.17% of 80,377 articles published in the field. The areas of surgery studied were analgesia 65 (49%), antibiotics 17 (13%), extracorporeal membrane oxygenation (ECMO) 9 (7%), gastrointestinal, burns, oncology, minimally invasive surgery, vascular access, congenital anomalies, and trauma (each <5%). Only 16 (12%) trials compared 2 surgical therapies, 9 (7%) compared a medical versus a surgical therapy, and 109 (81%) compared 2 medical therapies in surgical patients. Fourteen (10%) RCTs were funded by peer-reviewed agencies. Only 17 (13%) RCTs included a biostatistician as an author or a consultant. Trial design included calculation of sample size and statistical power in 21 (16%) RCTs. Method of randomization was reported in only 51 (38%). The test statistic and observed probability value was reported in 15 (11%). CONCLUSIONS: Clinical trials are used infrequently to answer questions related to pediatric surgery. When RCTs are utilized, they often suffer from poor trial design, inadequate statistical analysis, and incomplete reporting. Pediatric surgery could benefit from increased expertise, funding, and participation in clinical trials. PMID- 11479853 TI - Evaluation of pediatric surgery information on the Internet. AB - BACKGROUND/PURPOSE: Increasing numbers of parents use the Internet to obtain information about their child's medical diagnosis. Unfortunately, this information is not screened or regulated. The authors sought to evaluate the information available on the Internet regarding intersex anomalies as a representative pediatric surgical diagnosis. METHODS: Six search engines were searched for ambiguous genitalia. The first 30 sites on each search engine were reviewed. Sites were reviewed to identify information that did not conform to accepted recommendations for evaluation and treatment. RESULTS: Searches for ambiguous genitalia and synonyms found 0 to 44,471 sites per search engine. Of the 300 sites reviewed, only 45 represented 8 unique sites offering medical information. Five of these sites conformed to recommendations in 2 standard pediatric surgery texts, whereas 3 offered misleading information or information that did not conform to text recommendations. Of the total 300 sites, only 5 (1.6%) of accessible pages offered appropriate medical information to parents. CONCLUSIONS: Parent-oriented information regarding intersex anomalies is difficult to find on the Internet. Over one third of sites containing medical information failed to conform to standard pediatric surgical recommendations for treatment. Pediatric surgeons should help parents effectively use Internet information. PMID- 11479854 TI - The optimal approach for management of metachronous hernias in children: a decision analysis. AB - PURPOSE: Up to 30% of children undergoing unilateral hernia repair will later get a hernia on the contralateral side that requires repair. Three approaches have been used to address the potential for development of a metachronous hernia: (1) observation and repair of a contralateral hernia only if it later becomes apparent, (2) routine contralateral groin exploration, and (3) laparoscopy to evaluate the contralateral groin for a potential hernia. The purpose of this study was to use decision analysis to determine which approach resulted in the lowest morbidity, mortality, and cost. METHODS: A decision tree was constructed for the management of metachronous hernias that accounted for the occurrence of both nonincarcerated and incarcerated metachronous hernias. Baseline values were estimated from recent reports in the literature. Total charges for each approach were obtained from available hospital records and were used to estimate cost. RESULTS: Observation was favored over laparoscopy and laparoscopy over routine exploration with respect to preventing spermatic cord injury and preserving future fertility. Although a second operation may be required when observation is used, this approach was associated with only a small increase in anesthesia related complications (1 in 17,847), cardiac arrests (1 in 62,500), and death (1 in 312,500). Although observation was the favored approach with respect to cost, laparoscopy was less expensive when the expected incidence of metachronous hernias was high. CONCLUSIONS: Observation is the preferred approach to metachronous hernias because it results in the lowest incidence of injury and cost for most patients and is associated with a minimal increase in anesthesia related morbidity and mortality. Laparoscopy may be advantageous for patients at high risk for development of a contralateral hernia. As a strategy for preventing metachronous hernias without consideration for injury or cost, routine exploration should be limited to situations in which laparoscopy cannot be performed because of small patient size or a preference for spinal anesthesia. PMID- 11479855 TI - Kasai portoenterostomy: differences related to race. AB - PURPOSE: The aim of this study was to evaluate the Kasai portoenterostomy in African-American and white children with respect to differences in presentation and outcome. METHODS: A retrospective review of all children with biliary atresia who underwent a portoenterostomy at our institution over the last 15 years (n = 63) was performed. Sex, age at the time of Kasai, preoperative laboratory tests, success rates (defined as postoperative total serum bilirubin < or = 2.0 mg/dL), and survival rate were recorded. Differences between African-American (AA; n = 30) and white (W; n = 33) children were analyzed. Long-term follow-up was available on 59 of 63 patients. RESULTS: Sixty-three percent of all patients (40 of 65) were girls, and 48% were AA (30 of 63). A higher percentage of AA children (73%) were girls than were white children (55%), although this difference did not achieve statistical significance. African-Americans underwent portoenterostomy at a later age, had higher alkaline phosphatase levels, and higher AST. These differences were statistically significant. Preoperative ALT, total bilirubin level, and GGTP levels all were greater in African-Americans, although these differences did not achieve statistical significance. There was a trend toward decreased success and survival rate, although these results also were not statistically significant. CONCLUSIONS: African-Americans underwent primary therapeutic intervention for biliary atresia at an older age than white children with a trend toward less favorable results. These differences related to race may be attributed to greater difficulty in diagnosing jaundice or poorer access to health care in this patient population. Increased effort at identifying biliary atresia in AA children may lead to earlier diagnosis and treatment and improved outcomes. PMID- 11479856 TI - Venoarterial versus venovenous extracorporeal membrane oxygenation in congenital diaphragmatic hernia: the Extracorporeal Life Support Organization Registry, 1990 1999. AB - BACKGROUND/PURPOSE: Venoarterial (VA) extracorporeal membrane oxygenation (ECMO) traditionally has been the mode of support used in congenital diaphragmatic hernia (CDH). A few studies report success using venovenous (VV) ECMO. The purpose of this study is to compare outcomes in CDH patients treated with VA and VV. METHODS: The authors queried the Extracorporeal Life Support Organization Registry for newborns with CDH treated with ECMO from January 1, 1990 through December 31, 1999. They analyzed the pre-ECMO data, ECMO course, and complications. RESULTS: VA was utilized in 2,257 (86%) and VV in 371 (14%) patients. The pre-ECMO status was similar, with greater use of nitric oxide, surfactant, and pressors in VV. Survival rate was similar (58.4% for VV and 52.2% for VA, P =.057). VA was associated with more seizures (12.3% v 6.7%, P =.0024) and cerebral infarction (10.5% v 6.7%, P =.03). Sixty-four treatments were converted from VV to VA (VV-->VA). Survival rate in VV-->VA was not significantly different than VA (43.8% v 52.2%, respectively; P =.23). VV-->VA and VA patients had similar neurologic complications. CONCLUSIONS: CDH patients treated with VV and VA have similar survival rates. VA had more neurologic complications. The authors identified no disadvantage to the use of VV as an initial mode of ECMO for CDH, although some infants may need conversion to VA. PMID- 11479857 TI - Graft survival in pediatric liver transplantation. AB - BACKGROUND/PURPOSE: Liver transplantation is standard therapy for children with a variety of liver diseases. The current shortage of organ donors has led to aggressive use of reduced or split grafts and living-related donors to provide timely liver transplants to these children. The purpose of this study is to examine the impact of these techniques on graft survival in children currently treated with liver transplantation. METHODS: Data were obtained on all patients less than 21 years of age treated with isolated liver transplants performed after January 1, 1996 in an integrated statewide pediatric liver transplant program, which encompasses 2 high-volume centers. Nonparametric tests of association and life table analysis were used to analyze these data (SAS v 6.12). RESULTS: One hundred twenty-three children received 147 grafts (62 at the University of Florida, 85 at the University of Miami). Fifty-two (36%) children were less than 1 year of age at time of transplant, and 80 (55%) were less than 2 years of age. Patient survival rate was identical in the 2 centers (1-year actuarial survival rate, 88.4% and 87.1%). Twenty-five (17%) grafts were reduced, 28 (19%) were split, 6 were from living donors (4%), and 88 (60%) were whole organs. One-year graft survival rate was 80% for whole grafts, 71.6% for reduced grafts, and 64.3% for split grafts (P =.06). Children who received whole organs (mean age, 6.1 years) were older than those who received segmental grafts (mean age, 2.5 years; P <.01). Multifactorial analysis suggested that patient age, gender, and use of the graft for retransplant did not influence graft survival, nor did the type of graft used influence patient survival. CONCLUSIONS: The survival rate of children after liver transplantation is excellent independent of graft type. Use of current techniques to split grafts between 2 recipients is associated with an increased graft loss and need for retransplantation. Improvement in graft survival of these organs could reduce the morbidity and cost of liver transplantation significantly in children. PMID- 11479858 TI - A meta-analysis of peritoneal drainage versus laparotomy for perforated necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: Both primary peritoneal drainage (PPD) and laparotomy (LAP) are used widely for treatment of perforated necrotizing enterocolitis (NEC). Published reports include only anecdotes and small series. The authors used techniques of meta-analysis to determine which treatment is most effective. METHODS: The authors identified published studies reporting surgical treatment of NEC from January 1, 1978 to December 31, 1999; there were 10 studies (n = 475). The authors were contacted and all available raw patient data for use in meta analysis (n = 190) were obtained. The authors used logistic regression to determine the relative survival rate after PPD and LAP, controlling for the effect of gestational age and institution. RESULTS: The combined probability of survival in the 10 published studies did not show an advantage for PPD (55%) or LAP (67%; P =.27). When the authors corrected for the effect of birth weight on survival rate, they still did not observe a difference (P =.67). A marked bias in treatment assignment was found with smaller babies undergoing PPD than LAP (931 g versus 1,615 g, respectively; P =.0004). Analysis of raw data showed an even greater bias in treatment assignment. The authors found increased survival rate for LAP versus PPD (62.3% v 35.6%; P =.0009). However, a logistic regression model could not overcome the bias in assignment of patients with a much higher expected mortality rate to PPD. CONCLUSIONS: Using currently available data, it is not possible to determine whether PPD or LAP is superior. Bias in treatment assignment precludes conclusions regarding comparative survival. Only a randomized trial will determine which operation is best for the treatment of perforated NEC. PMID- 11479859 TI - Acute appendicitis in children: the importance of family history. AB - BACKGROUND/PURPOSE: Acute appendicitis is common, frequently atypical, challenging, and still associated with significant morbidity. Despite major technologic advances, appendicitis remains a primarily clinical diagnosis. Therefore, no relevant anamnestic information should be overlooked. Surprisingly, the relationship between heredity and appendicitis is seldom considered. Because of the potential clinical importance of the family history, the authors addressed this question prospectively over a 52-month period in a practice that includes the majority of pediatric patients with appendicitis in the region. METHODS: Family histories were obtained in a standardized manner, focusing on first-degree relatives. Children with incomplete family information were excluded. Patients (ages 2(1/2) to 19 years) were divided into 3 groups: group A, children who underwent an appendectomy (n = 166); group B (first control), children who presented with an acute abdomen and suspected appendicitis but did not undergo an appendectomy (n = 117); group C (second control), children who were seen in the practice for unrelated conditions (n = 141). RESULTS: A positive parental history was obtained from 59 patients (36%) in group A, 24 patients (21%) in group B, and 20 patients (14%) in group C, and the odds ratios (ORs) were 2.0 (P =.035) and 2.9 (P <.001) for groups A versus B and A versus C, respectively. Of the 13 patients whose sibling had had acute appendicitis, 9 were in group A versus 2 each in groups B and C, and the OR for any family history (siblings, parents) in groups A versus B was 1.9 (P =.028) and for groups A versus C was 2.9 (P < 0.001). Appendicitis was histologically confirmed in 93% of children in group A. CONCLUSIONS: Heredity is a significant factor in pediatric patients who have appendicitis. Children who have appendicitis are twice as likely to have a positive family history than are those with right lower quadrant pain (but no appendicitis) and almost 3 times as likely to have a positive family history than are surgical controls (without abdominal pain). Because of its potential value in changing the threshold for intervention, a careful family history should be obtained for every child in whom acute appendicitis is suspected. PMID- 11479860 TI - Fecal continence in patients having undergone posterior sagittal anorectoplasty procedure for a high anorectal malformation improves at adolescence, as constipation disappears. AB - BACKGROUND/PURPOSE: Constipation is a major complication in patients who have undergone posterior sagittal anorectoplasty (PSARP) operation for a high anorectal malformation. Overflow incontinence is the main cause of fecal soiling in these patients. The aim of this study was to outline the natural history of constipation in patients with high anorectal malformations and relate this to the functional outcome at the end of the patient's growth period. METHODS: The study group consisted of 22 pubertal or postpubertal patients (median age 15; range, 13 to 25) with high or intermediate anorectal malformations repaired by PSARP procedure. The patients have been followed-up since birth. Constipation was defined as a need to use medical treatment or diet to ensure bowel emptying. Continence was classified as follows: grade 1, no soiling in any circumstances; grade 2, staining less than once a week, no fecal accidents; grade 3, staining more than once a week, no fecal accidents; grade 4, daily soiling or accidents, need for regular enemas, or the antegrade colonic enema procedure. All patients underwent anorectal manometry and magnetic resonance imaging of the spine and spinal cord. RESULTS: At the time of the study 2 (9%) of the 22 study group patients had constipation, but 15 (68%) had been constipated before puberty. Eleven patients (50%) were fully continent (grade 1) without constipation. Six of those had a history of constipation associated soiling. Three patients (14%) had occasional staining (grade 2) and no constipation. Two of them had been constipated with significant soiling before the onset of puberty. In the 5 (22%) patients with frequent staining (grade 3) the degree of soiling had decreased after the disappearance of constipation. Two of the 3 patients with poor outcome (grade 4) require regular enemas for recalcitrant constipation. Spinal cord anomalies were detected in 4 and abnormal sacrum in 15 patients. Of the anorectal manometric parameters, only the force of voluntary sphincter squeeze correlated with the functional result. CONCLUSION: In the majority of patients who underwent PSARP procedure for high anorectal malformation, constipation disappears at adolescence, and this is associated with improved fecal continence outcome. PMID- 11479861 TI - Expression of neuropeptides in normal and abnormal appendices. AB - BACKGROUND/PURPOSE: Increased neuroproliferation in the appendix associated with an increase in substance P (SP), vasoactive intestinal polypeptide (VIP), and growth-associated protein-43 (GAP-43) has been documented in appendices of adults with acute right lower quadrant (RLQ) abdominal pain and absence of gross or histologic signs of appendiceal inflammation. The authors tested whether these findings were present in children with RLQ pain and a normal appendix. METHODS: Immunohistochemistry staining of paraffin-embedded appendices was performed with GAP-43, VIP, and SP. The positive control group included appendices with acute inflammation (group I, n = 5); the negative control group included appendices removed incidentally (group II, n = 5); and the experimental group included appendices from children suspected to have acute appendicitis without histologic signs of inflammation (group III, n = 9). RESULTS: Group I: VIP was strongly expressed in the nerve plexuses. The lamina propria and muscularis showed absent or minimal VIP expression. SP staining was strong in all plexuses and was moderate to strong in the muscularis. SP expression in the epithelium and lamina propria was difficult to quantify secondary to inflammation. Group II: VIP expression was essentially undetectable in the epithelium, lamina propria, and muscularis, and was moderate in the nerve plexuses. Mild SP staining was detected in the nerve plexuses of most specimens, and absent to mild staining was found in the epithelium and muscularis. However, one specimen strongly expressed SP in all layers. Group III: VIP expression was moderate to strong in the lamina propria and muscularis of nearly all specimens, and strong expression was found in all nerve plexuses. All but one specimen strongly expressed SP in plexuses. There was moderate to strong expression of SP in the epithelium, lamina propria, and muscularis in over 50% of specimens. The immunostaining for GAP-43 was very weak and nonspecific and did not help discriminate between the 3 study groups. CONCLUSIONS: Increased neuroproliferation in the lamina propria and muscularis was evident in patients with abdominal pain and normal appendices compared with appendices removed incidentally. The VIP and SP expression in these patients was similar or higher than that observed in patients with acute inflammation on histology. PMID- 11479862 TI - Antegrade continence enemas in the treatment of slow-transit constipation. AB - BACKGROUND/PURPOSE: Children with anorectal dysfunction can now be treated by antegrade continence enema (ACE), as described Malone et al. Those with idiopathic constipation, however, are not thought to be suitable for this treatment. Over 150 children attend the authors' department with proven slow transit constipation (mostly proven on nuclear transit/X-ray study), and the authors reviewed the outcome in the 40 of these who have had the ACE procedure. Families completed a questionnaire and attended interview with an independent assessor. METHODS: Of the 40 patients, 32 patients were assessed. Follow-up ranged from 3 to 54 months (median, 18 months) and age ranged from 5 to 17 years (median age, 10 years). Three of 32 stomas were no longer in use. Frequency of soiling was reduced significantly in 20 patients, and a further 6 patients were clean (P <.01). Abdominal pains were relieved significantly (P <.05), and appetite and mood improved. RESULTS: Stomal complications were frequent, (stenosis in 16 of 29, mucus leak in 20 of 29, fecal leak 3 of 29, catheter related pain in 20 of 29). Slow evacuation (12 of 29) and pain with enema (17 of 20) also were common. CONCLUSION: Malone appendicostomy does improve the well being of patients with slow transit constipation, but the advantages are less dramatic than in children with normal motility. PMID- 11479863 TI - Technical assessment of porcine enteroenterostomy performed with ZEUS robotic technology. AB - BACKGROUND/PURPOSE: Limitations of minimally invasive pediatric surgery include the inability to perform precise anastomoses of 2 to 15 mm. Robotic technology facilitates the performance of endoscopic microsurgical procedures. This study examined the technical feasibility of performing an enteroenterostomy in piglets utilizing ZEUS robotic technology. METHODS: Ten piglets (6.5 to 8.5 kg) underwent enteroenterostomy. Standard laparoscopic techniques were used in the control group (n = 5), and ZEUS robotic technology was used in the experimental group (n = 5). AESOP controlled the camera in both groups. Anesthesia time; surgery time; robotic set-up time; and anastomotic time, patency, diameter, and integrity were compared. RESULTS: No statistical difference existed between the means of the control and experimental groups for anesthesia time (176.0 v 154.0 minute; P =.63), surgery time (143.0 v 139.2 minute; P =.92), anastomosis time (109.4 v 93.0 minutes; P =.56), AESOP set-up time (4.2 v 7.0 minutes; P =.51), and anastomotic diameter (7.062 v 7.362 mm; P =.62). All anastomoses were patent without narrowing. The ZEUS cases averaged 14 minutes faster than the standard laparoscopic cases, even with the ZEUS set-up time included. CONCLUSIONS: These data supports the hypothesis that robotic-assisted enteroenterostomy is technically feasible. ZEUS robotic technology will potentially play an important role in expanding the applications of minimally invasive pediatric surgery. PMID- 11479864 TI - Is end-tidal CO2 an accurate measure of arterial CO2 during laparoscopic procedures in children and neonates with cyanotic congenital heart disease? AB - BACKGROUND/PURPOSE: Children with cyanotic congenital heart disease (CCHD) may require laparoscopic procedures. There are no data on the ability of capnography to predict arterial carbon dioxide concentrations (PaCO2) in patients with CCHD during pneumoperitoneum. METHODS: Seven patients (age 1 to 35 months) with CCHD undergoing laparoscopic Nissen fundoplication are presented. Standard general endotracheal anesthesia was administered. The operations were performed either by or in consultation with the cardiac anesthesia team. During each case, concomitant PaCO2 and end-tidal carbon dioxide (ETCO2) measurements were made via an arterial line and capnograph before and after insufflation of the abdomen. The PaCO2-ETCO2 gradients before and during pneumoperitoneum were then compared using a paired Student's t test. RESULTS: There was a statistically significant increase in the PaCO2-ETCO2 gradient (5.7 v 13.4) after insufflation compared with baseline (P <.015). CONCLUSIONS: These preliminary results show that ETCO2 is not a reliable monitor of PaCO2 in patients with CCHD undergoing laparoscopic procedures. The authors feel that close monitoring, including arterial blood gas measurements, and an experienced anesthesia team are necessary to perform laparoscopic procedures in patients with CCHD. PMID- 11479865 TI - Continuous intrapulmonary distension with perfluorocarbon accelerates lung growth in infants with congenital diaphragmatic hernia: initial experience. AB - BACKGROUND/PURPOSE: The authors have shown previously in an animal model that neonatal lung growth can be accelerated by continuous intrapulmonary distension with a perfluorocarbon (PFC). The authors now describe a preliminary clinical experience with this therapeutic concept in a select group of infants with congenital diaphragmatic hernia (CDH). METHODS: Neonates with very high predicted mortality rate caused by CDH had their lungs completely filled with PFC while on extracorporeal life support (ECLS); (n = 5). A continuous positive pressure of 7 to 10 cm H2O was maintained via the endotracheal tube for 3 to 7 days (mean, 5.6 +/- 0.87 days). The areas of both lungs (L) then were measured daily from digitized chest x-rays and divided by the area of the corresponding L1 vertebrae (V), to create an L/V index, so as to control for variable roentgenographic techniques. Immediately after removal of PFC, blood gas data were collected off ECLS. RESULTS: At the end of continuous pulmonary distension, all patients showed improvements in oxygenation and ventilation. The ipsilateral lungs showed significant increase of the L/V index with time (P =.003) and of L/V's daily change (P <.0001), suggesting accelerated lung growth. Overall survival rate was 40% (2 of 5). Of the 3 patients that had 7 days of distension, 2 survived. CONCLUSIONS: Continuous intrapulmonary distension with PFC for up to 1 week accelerated ipsilateral lung growth, improved gas exchange, and increased survival of CDH infants with profound pulmonary hypoplasia marooned on ECLS. Additional trials of PFC-based pulmonary distension in similar infants are warranted. PMID- 11479866 TI - The management of prenatally diagnosed choledochal cysts. AB - BACKGROUND: Although congenital biliary tree abnormalities are uncommon in the United States, more are being diagnosed antenatally with improved imaging techniques. METHODS: To determine the prognosis of prenatally diagnosed biliary disease, the authors reviewed the treatment of 3 children who had biliary cystic lesions found during routine prenatal ultrasonography. RESULTS: All 3 children were born at term. They had elevated bilirubin levels, and postnatal ultrasound scans confirmed the presence of a biliary cystic mass. They underwent exploration within 2 weeks of life. At operation, 2 children were found to have biliary atresia with a cystic biliary lesion. They underwent Kasai procedures and are doing well at 5 and 9 months of age. The third child was found to have a type I choledochal cyst and malrotation at exploration. He underwent cystectomy with Roux-en-Y hepaticojejunostomy and a Ladd's procedure and is doing well at 3 years of age. CONCLUSIONS: Prenatally diagnosed biliary cysts represent a different spectrum of disease than those diagnosed later in life. The sequelae of biliary disease start before birth, and early operation may be necessary to achieve a good outcome. Because it is impossible to distinguish between choledochal cysts and biliary atresia on antenatal ultrasound scan or magnetic resonance imaging, children with presumed choledochal cysts should undergo early exploration to rule out potential biliary atresia. Excellent outcome is possible with early operation in the absence of severe associated anomalies. PMID- 11479867 TI - Outcome of prenatally diagnosed solid fetal tumors. AB - BACKGROUND/PURPOSE: In the last 10 years, the ability to diagnose fetal tumors in the prenatal period has improved greatly because of technical advances in imaging. Early diagnosis and determination of tumor may affect prognosis. METHODS: The authors retrospectively reviewed the records of 1316 fetuses who underwent sonographic evaluation for congenital defects at University of California-San Francisco over a 6-year period. Of these, 16 had fetal tumors and were followed up at our institution. There were solid or predominantely solid with small cystic component masses in one of 3 locations: cervical, mediastinal, or abdominal. Excluded from our study were those fetuses with either sacrococcygeal teratoma, congenital cystic adenomatoid malformation of the lung, or ovarian cyst, because these defects have been extensively reviewed elsewhere. In addition, masses that were primarily cystic also were excluded. Data collected included diagnosis, gestational age at diagnosis and at delivery, mode of delivery, fetal and neonatal survival, and disease confirmation. RESULTS: Of the 16 fetuses, 4 had mediastinal tumors: 2 with pericardial teratomas (both of whom died in utero) and 2 with cardiac rhabdomyomas (1 died; the other presented tuberous sclerosis and is alive at 2 years of age). Four patients had cervical tumors (3 died; 1 survived and is alive and well), and 8 had abdominal tumors (3 with liver tumors, 4 with a left adrenal mass, and 1 with retroperitoneal teratoma). All eight patients with an abdominal tumor are alive and well. CONCLUSIONS: Fetal tumors are rare, and the prognosis seems to depend on their location and size. Although easier to detect, cervical and mediastinal tumors have a worse prognosis. Abdominal masses are more difficult to detect but have a better prognosis. PMID- 11479868 TI - Laparoscopic Heller myotomy and Dor fundoplication for esophageal achalasia in children. AB - BACKGROUND/PURPOSE: In the past, surgical treatment in achalasia usually has been reserved for patients whose dysphagia does not respond to pneumatic dilatation. The success of minimally invasive myotomy, however, has resulted in a shift in practice in adult patients, whereby laparoscopic surgery is becoming preferred as primary treatment by most gastroenterologists and surgeons. The aim of this study was to assess the efficacy of laparoscopic Heller myotomy and Dor fundoplication for esophageal achalasia in children. METHODS: Thirteen patients with esophageal achalasia (median age, 15 years; 6 boys and 7 girls; median duration of symptoms, 24 months) underwent laparoscopic Heller myotomy and Dor fundoplication between 1996 and 1999. Two patients had been treated previously by pneumatic dilatation, and 1 patient had received intrasphincteric Botulinum toxin injections. RESULTS: Median duration of the operation was 130 minutes. The patients were fed after an average of 33 hours, and they all left the hospital within 2 days. At a median follow-up of 19 months, there was no residual dysphagia in any patient. CONCLUSIONS: Laparoscopic Heller myotomy and Dor fundoplication were effective and safe for children with esophageal achalasia. Hospital stay and recovery time was short, and the functional results were excellent. These data support the notion that laparoscopic Heller myotomy should become the primary treatment of esophageal achalasia in children. PMID- 11479869 TI - Staged reconstruction for middle aortic syndrome. AB - BACKGROUND/PURPOSE: Middle aortic syndrome is a rare condition that involves narrowing of the abdominal aorta and its visceral branches. The authors propose staged vascular repair to minimize renal ischemia and facilitate use of native arterial tissue for reconstruction. METHODS: Three adolescents (age 8(1/2), 12(1/2), 13(1/2)) presented with severe hypertension. Subsequent evaluation showed coarctation of the abdominal aorta extending above the celiac axis. All 3 patients had bilateral renal artery stenoses. There also were tight stenoses of the celiac or superior mesenteric arteries. In the first stage the right renal artery stenosis was relieved. In the youngest patient, this was accomplished by balloon angioplasty. However, in the other 2, right renal autotransplantation was performed to the right iliac vessels using end-to-side anastomoses of the renal artery and vein. Cold perfusion was used. The second stage was performed 2 to 5 months later via a thoracoabdominal approach in 2 patients. A Dacron tube graft was utilized from above the coarctation to the iliac bifurcation. The left renal arteries were detached and anastomosed end to side to the bypass graft. In 1 child there were actually 3 separate renal arteries that required reimplantation. In the youngest patient the aortic narrowing was relieved by a long Dacron patch aortoplasty and interposition of an internal iliac artery graft to the left renal artery. RESULTS: All 3 patients recovered well and returned to full activities. There was no measurable rise of BUN or serum creatinine postoperatively. Postoperative renal scans showed good renal perfusion bilaterally. Follow-up results 2 to 10 years later continue to show well functioning reconstructions. CONCLUSION: A staged approach is an effective reconstruction for children with middle aortic syndrome which minimizes risk to renal function. PMID- 11479870 TI - Transplantation of newborn esophagus: an experimental study. AB - PURPOSE: The aim of this study was to see if allogeneic transplantation (Tx) of newborn esophagus can create viable esophageal tissue that may be used for treating long gap esophageal atresia. METHODS: Specimens of thoracic esophagus from newborn Brown-Norway rats, each were transplanted into a pouch created in the distal omentum of 5-week-old Lewis rats. In group I no immunosuppressant was used. FK-506 was used in group II (0.2 mg/kg), group III (0.6 mg/kg), and group IV (1.2 mg/kg) until a predetermined day of graft harvesting (1, 2, 3, 4, 5, 6, and 8 weeks after Tx). FK-506 was used for only 2 weeks in group V (0.6 mg/kg), and group VI (1.2mg/kg), and transplanted esophageal grafts were harvested 1, 2, 3, and 4 weeks after cessation of 2 weeks course FK-506. Syngeneic esophagus transplants were used as controls. All grafts were examined by H&E staining to assess graft viability and degree of rejection. RESULTS: Each successfully transplanted esophagus appeared macroscopically as a tube like mass. Each graft could be mobilized to the thoracic cavity, because of the long omental pedicle. Graft survival in the control group was 100%. Rejection was observed in all grafts from groups I, II, V, and VI. In contrast, grafts from groups III and IV showed only minimal or no rejection. There was no evidence of side effects of FK 506 in rats in groups III and IV, except significantly slower weight gain compared with controls (P <.05). CONCLUSIONS: FK-506 successfully prevented rejection, although immunologic tolerance was not achieved. These observations suggest that the authors' procedure has the potential to produce viable esophageal tissue that could be a new option for treating long gap esophageal atresia. PMID- 11479871 TI - Thyroid alar cartilage laryngotracheal reconstruction for severe pediatric subglottic stenosis. AB - BACKGROUND/PURPOSE: Laryngotracheoplasty has become an accepted treatment alternative for subglottic stenosis. However, the best autogenous material for laryngotracheoplasty remains controversial. Autogenous superior thyroid alar cartilage (TAC) has been used successfully in single stage laryngotracheal reconstruction in children with subglottic stenosis. METHODS: This is a retrospective study of 6 children (mean age, 16.6 months) undergoing TAC graft laryngotracheoplasty between September 1995, and June 1999. Two children had immediate tracheal intubation for congenital subglottic stenosis. Four others had previous tracheostomy: 3 for severe postintubation subglottic stenosis and 1 for congenital subglottic stenosis. After an anterior cricoid split, a piece of TAC was sutured between the cut ends of the cricoid, with the graft perichondrium facing intraluminally. Endotracheal intubation was maintained postoperatively. RESULTS: Four children underwent successfully extubation 9 to 21 days (mean, 15.5 days) postoperatively. Two required tracheostomy, which was maintained because of severe laryngomalacia and laryngotracheobronchomalacia. One child was treated with CO2 laser because of symptomatic recurrence of the subglottic stenosis 3 weeks after the surgery; another required fundoplication for gastroesophageal reflux 12 months after laryngotracheoplasty. There were no donor site complications in any of the 6 cases. Repeat laryngoscopy and bronchoscopy showed a patent subglottic airway. All of them are without symptoms after a mean follow up of 26 months. CONCLUSIONS: (1) This preliminary experience indicates that the TAC graft technique is a viable option for laryngotracheal reconstruction; (2) the TAC graft has significant advantages, including a single operative incision and absence of donor-site morbidity. PMID- 11479872 TI - Tracheal resection and reanastomosis in the neonatal period. AB - BACKGROUND/PURPOSE: Severe congenital tracheal stenosis is rare. Most of these can be managed conservatively before elective repair. Focal tracheal stenosis has been treated with resection of the involved trachea and primary reanastomosis in older infants. The authors found no reports of repair of this lesion in neonates. Two patients are presented with severe respiratory failure on the first day of life that required extracorporeal life support (ECLS) who underwent successful tracheal resection and reanastomosis (TRR) during the first week of life. METHODS: A retrospective review was conducted. RESULTS: Both babies had severe pulmonary hypertension and carbon dioxide retention despite maximal therapy and were placed on ECLS shortly after transfer. One had an isolated stenosis of the upper trachea, and the other had agenesis of the right lung, esophageal atresia with tracheoesophageal fistula, and a tracheal stenosis at the end of a short trachea with a long, narrow left bronchus. Both underwent diagnostic studies and had surgical repair while on ECLS at day 3 and 7 of life without bleeding complications. They were weaned off ECLS 1 and 8 days after surgery. One patient was extubated and did well. The other was extubated transiently, but required a tracheostomy because of left mainstem bronchomalacia. Both are alive and well at 18 and 38 months of age, with no narrowing of the repairs. CONCLUSION: In the setting of severe respiratory failure requiring ECLS support, TRR can be performed safely and successfully in the neonate with focal tracheal stenosis. PMID- 11479873 TI - A simple technique for preventing bar displacement with the Nuss repair of pectus excavatum. AB - BACKGROUND/PURPOSE: The most common complication of the minimally invasive technique for repair of pectus excavatum (MIRPE) is bar displacement, which has been reported to occur in 9.5% of all cases, particularly in teenaged patients. The use of a lateral stabilizing bar has improved stability but has not eliminated the occurrence of this problem. The authors report a new technique added to the standard MIRPE that creates an additional third point of fixation of the pectus bar to prevent displacement. METHODS: The technique requires the simple placement, via a spinal needle, of a nonabsorbable suture next to the sternum, encircling a rib and the bar, using a single 3-mm stab wound and thoracoscopic guidance. The suture simply is buried under the skin. Since 1998, this technique has been applied to 20 patients who underwent MIRPE. RESULTS: The average age was 14 years; 80% were boys. Average operating time was 75 minutes, and all patients had thoracoscopy with the MIRPE. A lateral stabilizing bar also was used in 14 patients. Four patients had 2 struts placed. Average length of stay was 5.5 days. There were no early complications. Mean follow-up was 12 months. Bar displacement occurred in 1 patient early in the series in which an absorbable suture was used for fixation. One patient had a prolonged hospital stay of 7 days because of postoperative pain. CONCLUSIONS: This modification to the original technique of MIRPE creates a 3-point fixation system that minimizes the risk of bar shifting even in teenaged patients. It does not add any significant time or cost to the operation, and it is fairly simple to perform. The authors believe that this technique decreases the occurrence of bar displacement, and they recommend its use for all patients with pectus excavatum considered candidates for the Nuss repair. PMID- 11479874 TI - Life-threatening anomalies of the thoracic duct: anatomic delineation dictates management. AB - Congenital anomalies of the thoracic duct are rare, poorly characterized, and difficult to manage. The spectrum of pathophysiologic perturbations, presenting symptoms, radiographic findings, and interventions performed in 4 patients are shown. Accurate anatomic delineation of the malformation was only possible by direct injection contrast lymphangiography. Therapies tailored to address the anatomic aberrations included intralesional sclerotherapy, surgical excision and ligation, lymphovenous anastomosis, and omental interposition to interrupt dysfunctional collateral lymphatics to the lung. Accurate anatomic diagnosis of central lymphatic channel anomalies by contrast lymphangiography facilitates an individualized multidisciplinary approach to repair. PMID- 11479875 TI - Effective therapy of a vascular tumor of infancy with vincristine. AB - Vascular tumors are common in infancy, affecting as many as 10% of children. These lesions often follow a benign course, with an initial proliferative phase followed by spontaneous involution, and require no therapy. Others manifest explosive early growth and Kasabach-Merritt phenomenon, requiring therapeutic intervention. Occasionally, some bulky tumors threaten life or vision because of mass effect, also mandating intervention. Steroids are the mainstay of therapy, but often are ineffective. Interferon alpha (2a and 2b) has been used as second line therapy in cases of steroid failure. However, interferon therapy has been associated with a significant incidence of spastic diplegia. The authors present the case of a 3-month-old girl in whom respiratory distress secondary to tracheal compression developed. Magnetic resonance imaging and magnetic resonance angiography showed a large cervicothoracic lesion encasing the great vessels and displacing the airway. She did not display associated Kasabach-Merritt phenomenon. The lesion proved refractory to standard steroid therapy, but responded dramatically to 4 cycles of vincristine (0.05 mg/kg). Although this agent has been used in children with life-threatening Kasabach-Merritt phenomenon, this is the first time it has been described in the setting of compromised vital function. Vinca alkaloids recently have been shown to have potent antiangiogenic activities in experimental models. Given the low predicted incidence of side effects at this dose, vincristine used as an antiangiogenic agent may prove an attractive alternative therapy for patients with life threatening vascular tumors of infancy. PMID- 11479876 TI - Pyloric stenosis: new histopathologic perspective using confocal laser scanning. AB - BACKGROUND/PURPOSE: Idiopathic hypertrophic pyloric stenosis (IHPS) is a common infantile disorder characterized by enlargement of the pylorus and gastric outlet obstruction. Its complete etiology is still not fully understood, but recent research has focussed on abnormalities of nerve distribution. The authors used confocal laser scanning microscopy to perform 3-dimensional studies of pylorus biopsy specimens taken from cases of IHPS and present their findings. METHODS: Pylorus biopsy specimens obtained at pyloromyotomy from 6 infants with IHPS were studied using confocal microscopy and compared with 6 control pylorus biopsy specimens from patients without gastrointestinal disease. Biopsy specimens were pretreated to enhance nerve expression by using protein gene product 9.5 (PGP9.5) polyclonal antibody to identify enteric nerve system fibers. Double staining immunofluorescence was used to detect alpha smooth muscle actin (SMA), a smooth muscle marker. RESULTS: Control pylorus biopsy specimens showed many thin PGP9.5 positive nerve fibers in the circular and longitudinal muscle layers that communicated with each other to create a 3-dimensional meshlike network. Muscle cells stained by alpha SMA antibody were thin. In contrast, muscle cells from IHPS patients were fat and round. The PGP9.5 staining nerve fibers from IHPS patients formed numerous, thick, and contorted bundles that did not communicate. CONCLUSIONS: By using confocal laser microscopy the authors were able to identify abnormally thick contorted nerve bundles in the pyloric muscle layers of infants with IHPS. These anormal nerve bundles have not been described previously because of the limitations of 2-dimensional microscopy. The authors suspect that the etiology of IHPS may be related to these abnormal fibers. PMID- 11479877 TI - Selective neurotrophin deficiency in infantile hypertrophic pyloric stenosis. AB - BACKGROUND/PURPOSE: Increasing evidence suggests that the enteric nervous system is under the control of neurotrophins. Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5), promote differentiation, growth, and survival of various central and peripheral nervous system neurons. The biological effects of neurotrophins are mediated by the interactions with high-affinity tyrosine kinase receptors (TrkA, TrkB, TrkC). Recently, abnormalities of intramuscular innervation have been reported in infantile hypertrophic pyloric stenosis (IHPS). To further understand the reported abnormalities in pyloric innervation in IHPS, the authors analyzed the expression of Trk receptors and the neurotrophins content in IHPS. METHODS: Full thickness muscle biopsy specimens were obtained from 8 IHPS patients (age range, 23 to 41 days) at pyloromyotomy and from 8 age-matched controls without gastrointestinal disease at autopsy performed within 12 hours after death. Indirect immunohistochemistry was performed using ABC (Avidin Biotin peroxidase Complex) method with anti-Trk specific antibodies (A,B,C). Quantitative analysis was performed using sandwich-type ELISA for NGF, BDNF, NT-3, and NT-4/5. RESULTS: The intensity of staining of the myenteric plexus for TrkA, TrkB, and TrkC was similar among IHPS and controls. There was a lack of TrkA-positive nerve fibers in IHPS compared with controls. The quantity of total NGF, NT-3, and BDNF in IHPS was significantly lower than in controls. CONCLUSIONS: The reduced production of neurotrophins in IHPS may be responsible for the delay in the functional and structural maturation of pyloric innervation in IHPS. The lack of TrkA-positive nerve fibers in pyloric muscle may explain the abnormal intramuscular innervation in IHPS. PMID- 11479878 TI - Interleukin-12 transduced dendritic cells induce regression of established murine neuroblastoma. AB - BACKGROUND/PURPOSE: Interleukin 12 (IL-12) is a potent proinflammatory cytokine that enhances cytotoxic T lymphocytes (CTL) and natural killer (NK) cell activity. The goal of these experiments was to assess whether adenoviral-mediated IL-12 expression by dendritic cells (DC) could induce an antitumor immune response in a murine model of neuroblastoma. METHODS: Syngeneic A/J mice were inoculated subcutaneously with 1 x 10(6) cells from a murine neuroblastoma derived cell line (TBJ). Murine DC were transduced in vitro with adCMV-mIL-12, and 1 x 10(6) cells were injected intratumorally. Tumor growth in these mice was compared with control animals injected with enhanced green-filled protein (EGFP) transduced DC or saline. The role of CTL was evaluated through cytotoxicity assays. Immunohistochemical analysis of tumor, spleen, and lymph node was performed to characterize the behavior and fate of various populations of immune effector cells in these tissues. RESULTS: The tumors of mice injected with adIL 12 transduced DC all underwent complete regression over a 3-week period. Splenocytes isolated from mice 7 days after intratumoral injection of adIL-12 DC showed increased cytolytic activity relative to control animals in vitro. Immunohistochemistry of tumor and lymph tissue showed increased amounts of DC and T lymphocyte infiltration and a slight decrease in apoptosis relative to the control groups. CONCLUSIONS: Increased IL-12 production by DC induced a significant antitumor response in a poorly immunogenic murine neuroblastoma model. These results show the vital role of DC in the immunobiology of the disease, and that protection of these cells from tumor induced apoptosis is a critical aspect for immunotherapies treating this aggressive tumor. PMID- 11479879 TI - Intestinal neuronal dysplasia-like pathology in Ncx/Hox11L.1 gene-deficient mice. AB - BACKGROUND/PURPOSE: Ncx/Hox11L.1-deficient (Ncx-/-) mice specifically created by the authors had mega-ileo-ceco-colon (mega-ICC) with a caliber change in the proximal colon. The authors studied the nerve distribution in the bowel of these Ncx-/- mice to determine the cause of their bowel dysmotility. METHODS: Four-week old Ncx-/- mice (n = 10; 5 with mega-ICC, 5 without mega-ICC) were killed and the bowel harvested. Half of each specimen was snap frozen for AchE and NADPH diaphorase histochemistry, and the other half were fixed with 10% formalin for H&E staining and immunohistochemistry using PGP9.5 antibody (a marker for neurons), C-kit antibody (a marker for intestinal pacemaker cells), and stem cell factor antibody (a marker for C-kit ligand). Age-matched wild-type normal mice (n = 5) served as controls. RESULTS: In the ileum, cecum, and proximal colon from all Ncx-/- mice (irrespective of the association of mega-ICC), typical findings of human intestinal neuronal dysplasia (IND) ie, obvious hyperganglionosis in neuronal plexuses on PGP9.5 immunohistochemistry, ectopic ganglia in the mucosal and muscular layers on AchE histochemistry, and ghostlike ganglia on NADPH diaphorase histochemistry were found. Likewise, in normal caliber distal colon from these mice, the distribution of ganglion cells, C-kit, and stem cell factor was normal. In control specimens, there was no ectopic ganglia or hyperganglionosis. CONCLUSIONS: These findings suggest that the Ncx/Hox11L.1 gene is required for the proper innervation of the enteric nervous system in mice, and our deficient strain may be useful as a model for studying IND in humans. PMID- 11479880 TI - Serum concentration of adhesion molecules in postoperative biliary atresia patients: relationship to disease activity and cirrhosis. AB - BACKGROUND/PURPOSE: Biliary atresia (BA) is associated with progressive liver fibrosis, which may be mediated by immunologic abnormalities involving adhesion molecules. This study investigates the relationship between serum intercellular adhesion molecule-1 (sICAM-1), serum vascular cell adhesion molecule-1 (sVCAM-1), and the clinical and histologic severity of BA. METHODS: Serum ICAM-1 and VCAM-1 levels were measured by enzyme-linked immunosorbent assay in 35 patients with BA and 20 healthy controls. Standard liver function tests (LFTs), and frozen section liver biopsy specimens were used to determine liver status. On the basis of LFT results, the BA patients were classified into group I (n = 10; normal LFTs), group II (n = 15; elevated LFTs, anicteric), and group III (n = 10; elevated LFTs, icteric). Eight subjects in group II, and all subjects in group III had portal hypertension (PH). RESULTS: sICAM-1 levels were significantly elevated in group III (1760.0 +/- 717.5 ng/mL) compared with group II (555.1 +/- 199.4 ng/mL), group I (272.1 +/- 59.9 ng/mL) and controls (256.3 +/- 71.6 ng/mL). Although sVCAM-1 levels were significantly elevated in group III (1932.9 +/- 282.6 ng/mL) compared with group II (1054.3 +/- 297.0 ng/mL), group I (605.4 +/- 112.4 ng/mL), and controls (616.0 +/- 112.0 ng/mL; P <.001), there was no statistically significant difference between groups I, II, or controls. sVCAM-1 levels were elevated significantly in BA subjects in group II with PH (1253.0 +/- 245.1 ng/mL) compared with those who did not have PH (827.3 +/- 151.7 ng/mL; P <.01). PH did not affect sICAM-1 levels. There was strong expression of ICAM-1 and VCAM-1 in proliferating bile ductules, endothelial cells, and liver cells in group III compared with group II and controls. CONCLUSIONS: In BA, sICAM-1 and sVCAM-1 levels could be useful as markers of end-stage liver disease, with sVCAM 1 being more specific for PH. Induction of ICAM-1 and VCAM-1 may be an important factor in the development of cirrhosis. PMID- 11479881 TI - The utility of lung biopsy in recipients of stem cell transplantation. AB - BACKGROUND/PURPOSE: Pulmonary infiltrates in recipients of stem cell transplantation often present as diagnostic dilemmas. Although lung biopsy may establish the diagnosis of parenchymal disease, it remains unclear whether such a procedure results in a significant change in the patient's treatment and outcome. This study evaluates the efficacy of lung biopsy in recipients of stem cell transplantation. METHODS: The medical records of 15 stem cell transplant recipients who underwent 18 lung biopsies were reviewed. The indications for stem cell transplantation were leukemia in 10 patients, lymphoma in 2, histiocytosis in 1, neuroblastoma in 1, and Ewing's sarcoma in 1. The results of the lung biopsies were correlated to the clinical management and outcomes. RESULTS: The overall mortality rate was 67% (10 patients). Eight of the 9 patients who required mechanical ventilatory support at the time of lung biopsy died. The pathologic diagnoses were pneumonitis in 6 biopsies, fibrosis in 6, brochiolitis obliterans organizing pneumonia in 3, hemorrhage in 2, and infarction in 1. Therapy was changed in 1 patient who improved after a course of steroids for bronchiolitis obliterans organizing pneumonia. Lung biopsy cultures were positive in 6 patients but rarely resulted in changes in antibiotic therapy. CONCLUSIONS: Results of very few lung biopsies performed in stem cell transplant recipients redirected therapy. Furthermore, the ultimate outcome of these patients were not improved by the results of lung biopsies. PMID- 11479882 TI - Gene transfer to the embryo: strategies for the delivery and expression of proteins at 48 to 56 hours postfertilization. AB - BACKGROUND/PURPOSE: Although gene and protein transfer may potentiate the cure of genetic disease, current strategies involving fetal gene therapy remain nonfocal and confounded by the lack of imaging techniques and in vivo markers for precise gene transfer. METHODS: Fourteen white Leghorn chick eggs were incubated for 48 to 56 hours postfertilization until they reached stages 11 to 16, about 3 mm in size. In 7 chick embryos, a glass needle was placed at the midbrain/hindbrain level and 1 x 10(7) pfu of an adenovirus containing the green fluorescent protein (GFP) reporter gene was injected into the lateral head. In another 7 chicken embryos, colored agarose beads coated with Sonic hedgehog (Shh) protein were implanted at the level of the hindbrain under direct microscopy. The eggs were then sealed, incubated at 37 degrees C for 24 hours, and reimaged using fluorescent microscopy and confocal laser microscopy. RESULTS: At 24 hours postinjection, all embryos were alive and were imaged in vivo. Fluorescent microscopic imaging showed green fluorescence in the region of the injection site in all the embryos. In embryos that underwent bead placement, the beads were visualized under microscopy in the lateral hindbrain of all embryos, and the presence of the Shh protein was confirmed using fluorescein isothiocyanate (FITC) conjugated secondary antibody. CONCLUSIONS: This study shows that embryonic 3-mm chick embryos survive adenoviral transduction or agarose bead implantation in a focal manner in vivo and that this delivery results in production of imageable levels of protein. This may be used in mammalian systems, including humans, to introduce genes and proteins. PMID- 11479883 TI - Management of the high-risk pediatric burn patient. AB - BACKGROUND/PURPOSE: Inhalation injury, flame burn exceeding 30%, and age under 48 months all have been cited as independent risk factors for mortality; the combination of all 3 risk factors is unusual. The authors have experienced an overall reduction in mortality rate and chose to examine this high-risk group to define techniques useful in improving outcome in pediatric burns. METHODS: A review was done of children with all 3 risk factors over a recent 9-year interval. All were treated with a system of care emphasizing precise fluid repletion, early wound excision and closure, and avoidance of injurious pulmonary inflating pressures and concentrations of oxygen. Data are expressed as mean +/- SD. RESULTS: There were 26 children admitted with all 3 risk factors. Their average age was 2.1 +/- 1.1 years (range, 5 weeks to 3.7 years), and burn size was 61% +/- 21% (range, 30% to 98%) of the body surface. All required mechanical ventilation for an average of 28 +/- 4.5 days (range, 7 to 74 days). Two children underwent tracheostomy; all others were treated with protracted oral intubation. Inhaled nitric oxide (NO) was used in 3 children, all of whom were considered for extracorporeal membrane oxygenator (ECMO) support, although none went on to ECMO. Only 7 children (27%) never had any bacteremia. Ventilator-related pneumonia occurred in 8 children (31%). Total lengths of stay, including acute and rehabilitation hospitalizations, averaged 105 +/- 10 days (1.87 +/- 0.2; range, 0.66 to 4.8 days per percent burn). After exclusion of 1 child with a 98% third and fourth-degree burn, pre-hospital cardiac arrest, and anoxic brain injury who had support withdrawn at 6 hours, all children survived to discharge; 23 followed up in our clinic currently are alive and well with no overt residual respiratory insufficiency. CONCLUSION: A high rate of survival can be expected in young children with large burns and inhalation injury. PMID- 11479884 TI - Multislice helical CT depiction of Wilms' Tumor. PMID- 11479885 TI - Congenital cysts of the third and fourth pharyngeal pouches or pyriform sinus cysts. PMID- 11479887 TI - A case of invasive rectal adenocarcinoma in a 17-year-old girl with tuberous sclerosis complex. PMID- 11479888 TI - If infants rendered jaunice free (specifically with a normal bilrubin) after a Kasai. PMID- 11479889 TI - Successfully achieving orchiopexy in all their 8 patients using exclusively 2-mm optics and instrumentation. PMID- 11479891 TI - Molecular genetics of small cell lung carcinoma. AB - The etiology of small cell lung cancer (SCLC) is strongly tied to cigarette smoking, and now there is considerable information concerning molecular abnormalities involved in the pathogenesis of SCLC. Autocrine growth factors such as neuroendocrine regulatory peptides (eg, bombesin/gastrin-releasing peptide) are prominent in SCLC. Dominant oncogenes of the Myc family are frequently overexpressed in both SCLC and non-small cell lung cancer (NSCLC), while the K RAS oncogene is never mutated in SCLC but it is in 30% of NSCLCs. The most frequent genetic abnormalities involve tumor suppressor genes (TSGs). The TSG p53 is mutated in more than 90% of SCLCs and more than 50% of NSCLCs; the retinoblastoma TSG is inactivated in over 90% of SCLC but only 15% of NSCLCs, and p16, the other component of the retinoblastoma/p16 pathway, is almost never abnormal in SCLC but is inactivated in more than 50% of NSCLCs. The FHIT TSG is inactivated in 50% to 70% of all lung cancers. Recently, we completed a genome wide allelotyping study using approximately 400 polymorphic markers distributed at around 10 cM resolution across the human genome comparing SCLCs and NSCLCs, looking for all possible TSG sites by loss of heterozygosity. We found that, on average, 17 loci showed loss of heterozygosity in individual SCLCs and 22 for NSCLC, with an average size of loss of 50 to 60 cM, and an average frequency of microsatellite abnormalities of five per tumor. There were 22 different "hot spots" for loss of heterozygosity, 13 with a preference for SCLC, seven for NSCLC, and two affecting both. This provides clear evidence on a genome-wide scale that SCLC and NSCLC differ significantly in the TSGs that are inactivated during their pathogenesis. Acquired hypermethylation of the promoter region of key genes has become one of the most common mechanisms that tumors use to inactivate the function of tumor suppressor and other genes. We recently completed a study of tumor-acquired promoter hypermethylation for nine genes (p16, DAPK, MGMT, GSTP1, RAR beta, FHIT, ECAD, p14ARF, and TIMP1). We found differences in the frequency of RAR beta methylation (70% for SCLC and 40% for NSCLCs). Finally, we looked at the bronchial epithelium accompanying SCLC and NSCLC for the occurrence of clonal alterations using precise laser capture microdissection with subsequent allelotyping for polymorphic markers. In NSCLC, we frequently find clones of cells with molecular abnormalities in histologically affected epithelium (eg, carcinoma in situ, dysplasia, hyperplasia) and occasionally in normal-appearing epithelium in the cases of current or former smokers. In SCLC these histologic preneoplastic changes were minimal. However, in studies of histologically normal respiratory epithelium, we found a several-fold increased rate of allele loss in SCLC compared with NSCLC patients. Thus, the smoking-damaged histologically normal epithelium associated with SCLC appeared genetically scrambled and has incurred significantly more damage than the epithelium accompanying NSCLCs. We conclude that SCLC and NSCLCs do not differ significantly in the number of genetic alterations that occur. However, SCLCs do differ significantly from NSCLCs in the specific genetic alterations that occur. In addition, smoking-damaged bronchial epithelium accompanying SCLCs appears to have undergone significantly more acquired genetic damage than that accompanying NSCLCs. Future studies need to identify the specific genes involved at these multiple sites and determine if these provide new tools for early molecular detection and monitoring of chemoprevention efforts, and serve as specific targets for developing new therapies. Semin Oncol 28 (suppl 4):3-13. PMID- 11479892 TI - Combined-modality therapy for limited-stage small cell lung cancer. AB - While the International Union Against Cancer system can be used in staging small cell lung cancer (SCLC) patients, the staging distinction of "extensive stage" versus "limited stage" is commonly used. This system defines limited stage as the ability to encompass all known disease within a "reasonable" radiation field. The standard management of limited-stage SCLC is concurrent platinum-based chemotherapy with thoracic radiotherapy (RT). Total RT doses range from 40 to 55 gy in most trials. A recently completed randomized trial showed an advantage to twice-daily accelerated hyperfractionated RT over standard RT alone. Most current clinical protocols for limited-stage SCLC use the "involved field" technique of thoracic RT, in which the RT target volume includes the known extent of primary tumor and lymph node involvement and one additional nodal station. Large fields including more extensive elective nodal irradiation are discouraged. Semin Oncol 28 (suppl 4):14-22. PMID- 11479893 TI - Concurrent versus sequential radiotherapy for small cell lung cancer. AB - Two meta-analyses have shown that thoracic radiotherapy plus chemotherapy is superior to chemotherapy alone in the treatment of limited-stage small cell lung cancer. To minimize the likelihood of emergence of chemotherapy resistance, the optimal schedule for integration of the two modalities might be to introduce radiotherapy early in the course of treatment. The Cancer and Leukemia Group B trial performed from 1981 to 1984 failed to show an advantage for early concurrent chemoradiation. However, three recent controlled trials of thoracic irradiation timing (the National Cancer Institute of Canada trial, the Yugoslavian trial, and the Japan Clinical Oncology Group trial) showed that early radiotherapy is superior to delayed radiotherapy. These three trials used the combination of cisplatin/etoposide and showed that a long-term survival rate greater than 20% is achievable using early integrated chemoradiation. For the present, early concurrent administration of thoracic radiotherapy with a cisplatin/etoposide-based regimen should be standard therapy. Semin Oncol 28 (suppl 4):23-26. PMID- 11479894 TI - New drugs for chemotherapy-naive patients with extensive-disease small cell lung cancer. AB - To improve the survival of patients with small cell lung cancer, there is a need for new and effective agents to treat this disease. These agents include paclitaxel, docetaxel, topotecan, irinotecan, vinorelbine, gemcitabine, and amrubicin. In previously untreated small cell lung cancer patients the response rate for these effective drugs ranges from 27% to 79%. Median survival ranges from 6.6 to 12 months. The major toxicity for these agents is leukopenia. Studies are ongoing to evaluate the efficacy of combination chemotherapy using new agents either together or with other known effective drugs for the treatment of small cell lung cancer. Semin Oncol 28 (suppl 4):27-29. PMID- 11479895 TI - Alternatives to chemotherapy and radiotherapy in the treatment of small cell lung cancer. AB - Despite high initial response rates, most patients with small cell lung cancer (SCLC) relapse shortly after discontinuing therapy, and cure remains an elusive goal, even for patients with limited-stage disease. For this reason, investigators have turned to the evaluation of alternative treatment strategies for patients with this malignancy. Interferon has been evaluated in four trials of adjuvant therapy after response to chemotherapy for SCLC. Different interferon preparations were used, but none of the trials showed a significant prolongation of survival. Tumor vaccines against gangliosides that are expressed on almost all human SCLC cells have been recently developed. An anti-idiotypic monoclonal antibody against the GD3 ganglioside, BEC-2, is being evaluated after chemotherapy in SCLC patients in a European study. In North America, a bivalent ganglioside vaccine, BMS-248967, is under study at the phase II level. If results are promising, a phase III trial will be undertaken. Trials of anticoagulant therapy using heparin, warfarin, and aspirin were undertaken in the 1980s and early 1990s. Although one study of warfarin and another of heparin showed trends in favor of anticoagulant therapy, there has been little research over the past decade evaluating this form of treatment of SCLC. The matrix metalloproteinase inhibitors marimastat and BMS-275291 are under evaluation in SCLC. In all trials, the matrix metalloproteinase inhibitor is administered after chemotherapy and radiotherapy as adjuvant treatment. These trials are not yet mature and survival results are not available. Miscellaneous treatments, including monoclonal antibody therapy, tamoxifen, and growth factor inhibition have not yet been shown to have a role in the treatment of SCLC. Semin Oncol 28 (suppl 4):30-37. PMID- 11479896 TI - The importance of drug diversity in the treatment of small cell lung cancer. AB - Drug diversity is a fundamental concept guiding the assembly of combination chemotherapy regimens. At a basic level, the superiority of combination chemotherapy over monotherapy shows the importance of drug diversity. However, it has been difficult to show advantages of triplet regimens over doublet regimens in patients with extensive-stage small cell lung cancer. Among the potentially curable limited-stage small cell lung cancer patient population, a number of trials have suggested that more diverse regimens could be more active. However, the need to safely integrate thoracic irradiation with etoposide and cisplatin for limited-stage small cell lung cancer has decreased enthusiasm for additional drugs. The power of multiple traditional agents to overcome resistant clones is clearly limited. Clinical trials incorporating the taxanes and topoisomerase I inhibitors will explore the capacity of these more recently investigated agents to improve results. Semin Oncol 28 (suppl 4):38-42. PMID- 11479897 TI - Paclitaxel-based three-drug combinations for the treatment of small cell lung cancer: a review of the Sarah Cannon Cancer Center experience. AB - Between June 1993 and September 1999, 217 patients with small cell lung cancer (SCLC) entered three sequential phase II trials evaluating novel paclitaxel containing three-drug combination chemotherapy regimens. Patients with limited- or extensive-stage SCLC, no previous treatment, and Eastern Cooperative Oncology Group performance status 0 to 2 were eligible. Trials 1 and 2 evaluated the combination of paclitaxel, carboplatin, and etoposide; in the second trial, doses of paclitaxel and carboplatin were higher than in the first trial. Trial 3 evaluated the combination of paclitaxel, carboplatin, and topotecan. Patients with limited-stage disease received radiation therapy to the primary tumor site and mediastinum, beginning concurrently with the third course of chemotherapy. All patients received four courses of chemotherapy, administered at 21-day intervals. All three regimens were highly active and produced high response rates in both limited- and extensive-stage SCLC. Median survivals for regimens 1, 2, and 3 in extensive-stage patients were 8, 10, and 8.5 months, respectively. Median survivals in limited- stage disease were 16, 20, and 15 months, respectively. Although definitive comparisons of these regimens cannot be made on the basis of sequential trials, the higher-dose paclitaxel/carboplatin/etoposide regimen seemed superior; with this regimen, 4-year survival in limited-stage disease was 23%. Paclitaxel-containing three-drug regimens, as evaluated in these three phase II trials, were feasible and highly active in the first-line treatment of SCLC. Randomized trials will be necessary to definitively evaluate the efficacy of these regimens as compared with traditional platinum/etoposide combinations. Semin Oncol 28 (suppl 4):43-47. PMID- 11479898 TI - Treatment of relapsed small cell lung cancer. AB - Small cell lung cancer is a frequently relapsing tumor despite a high rate of response after first-line chemotherapy. For these patients, treatment at relapse depends on the response to first-line treatment, time off therapy, and the site of progression. For patients with a good performance status and a relapse more than 3 months after first-line treatment, the choice of chemotherapy at relapse is the same as initial therapy. In case of earlier progression, so-called non cross-resistant chemotherapy should be given. For symptomatic metastases in bone or brain, radiotherapy usually gives temporary palliation. For intrathoracic relapse and severe obstruction of main bronchus and/or superior caval vein, radiotherapy is a possibility if not given as part of first-line treatment. Semin Oncol 28 (suppl 4):48-52. PMID- 11479899 TI - Liver function tests during amoebic liver abscess formation in indomethacin treated hamsters. AB - Establishment of Entamoeba histolytica infection is facilitated through macrophage effector disruption by a prostaglandin E2 (PGE2)-mediated mechanism. Infection severity may be measured by weight of abscess formed. Indomethacin (Indo) treatment of infected hamsters reduced abscess weight by 30% at 7 days post-infection presumably by inhibition of PGE2. To explain reductions in abscess development by Indo treatment, we determined liver functionality in Indo-treated or untreated animals, either healthy or infected. Determinations of serum glutamic oxaloacetic (SGOT) and glutamic pyruvic (SGPT) transaminases, serum alkaline phosphatase (SAP), total serum protein (TSP), and bilirubinemia were done. SGOT, SGPT, and SAP activities showed a significant increase in their values by 600% at seven days post-infection in infected animals in both conditions; nonstatistical differences were found between animals treated or not. This increase did not correlate with the percentage of damage. Infected nontreated hamsters showed TSP levels 30% below normal group (P < 0.05). Infected Indo-treated hamsters had no significant differences compared to normal values. Infected nontreated animals showed an increase in bilirubin, particularly in indirect bilirubin, whereas infected Indo-treated hamsters showed total bilirubin values lower than normals (P < 0.05), with a decrease in direct bilirubin levels. Our results demonstrated that E. histolytica infection in hamsters produces similar abnormalities in liver function as it does in humans, and that the beneficial effect of Indo treatment on amoebic abscess development is not related with an improvement of liver functionality. PMID- 11479900 TI - Food chemical cues elicit general and population-specific effects on lingual and biting behaviors in the lacertid lizard Podarcis lilfordi. AB - Actively foraging lizards are capable of identifying prey using only chemical cues sampled by tongue-flicking, and the relatively few omnivorous and herbivorous lizards tested similarly can identify both animal and plant foods from chemical cues. Whether lizards that eat plants respond to cues specific to preferred plant types and whether there is geographic variability in responses to cues from various plants correlated with the importance of those plants in local diets is unknown. In three populations of an omnivorous lacertid, the Balearic lizard Podarcis lilfordi, we studied chemosensory sampling and feeding responses to chemical cues from plant and animal foods presented on cotton swabs. Each lizard population is endemic to one islet off the coast of Menorca, Balearic Islands, Spain. Lizards in all three populations discriminated chemical cues from plant and animal foods from control substances. Our results extend findings of prey chemical discrimination and plant chemical discrimination in omnivores, increasing confidence that correlated evolution has occurred between plant diet and chemosensory response to palatable plants. There were no consistent differences among populations in tongue-flicking and biting responses to stimuli from flowers of syntopic and allopatric plant species. The lizards may respond to cues indicative of palatability in a wide range of plant species rather than exhibiting strong responses only to locally available plant species. Nevertheless, tongue-flicking and biting frequencies varied among plant species, perhaps indicating food preferences. In addition, there were differences among populations in tongue-flick rates, latency to bite, and licking behavior. Licking was observed in only one lizard population as a response to floral chemicals from only one of the plants species tested, raising the possibility of a population specific linkage between identification of a particular plant species and performance of an appropriate feeding response. PMID- 11479901 TI - Is the unique negatively charged polypeptide of crayfish yolk HDL a component of crustacean vitellin? AB - The yolk protein of Cherax quadricarinatus contains six major high-density lipoprotein (HDL) subunits with the approximate molecular masses of 177, 155, 106, 95, 86, and 75 kDa, of which only the 106-kDa polypeptide is negatively charged. On the basis of their molecular weights, time of appearance and disappearance, their floating density and susceptibility to enzyme degradation (by a serine proteinase), these six HDL polypeptides were classified into two subgroups. One group comprises the higher-molecular-weight compounds above 106 kDa, and the other includes the lower-molecular-weight compounds up to 95 kDa. Other than being different from the lower-molecular-weight polypeptides, the negatively charged 106-kDa polypeptide was significantly different from members of its higher-molecular-weight group belonging to a different, less abundant, yolk protein as shown by HPLC separation. Immunological studies and peptide mapping in which the 106-kDa polypeptide did not show similarity to any of the other HDL components confirmed these differences. Moreover, the amino acid composition of the 106-kDa polypeptide was different from that of known vitellin from other crustacean species. This unique negatively charged polypeptide presents an enigma as it is known to be a secondary vitellogenic-related HDL polypeptide, immunolocalized in yolk globules; however, it is different to all the other HDL polypeptides, thus presenting the question whether it is indeed a component of "classical" crustacean vitellin. PMID- 11479902 TI - Expression of muscle-specific myosin heavy chain and myosin light chain 1 in the electric tissue of Electrophorus electricus (L.) in comparison with other vertebrate species. AB - Myosin light and heavy chains from skeletal and cardiac muscles and from the electric organ of Electrophorus electricus (L.) were characterised using biochemical and immunological methods, and compared with myosin extracted from avian, reptilian, and mammalian skeletal and cardiac muscles. The results indicate that the electric tissue has a myosin light chain 1 (LC1) and a muscle specific myosin heavy chain. We also show that monoclonal antibody F109-12A8 (against LC1 and LC2) recognizes LC1 of myosin from human skeletal and cardiac muscles as well as those of rabbit, lizard, chick, and electric eel. However, only cardiac muscles from humans and rabbits have LC2, which is recognized by antibody F109-16F4. The data presented confirm the muscle origin of the electric tissue of E. electricus. This electric tissue has a profile of LC1 protein expression that resembles the myosin from cardiac muscle of the eel more than that from eel skeletal muscle. This work raises an interesting question about the ontogenesis and differentiation of the electric tissue of E. electricus. PMID- 11479903 TI - Carbonic anhydrase isozyme distribution and characterization in metabolic fiber types of the dorsal levator muscle of the blue crab, Callinectes sapidus. AB - Carbonic anhydrase (CA) distribution and characterization were examined in white and light pink fibers of the dorsal levator muscle of the blue crab. White fibers were structurally and metabolically characterized as fast twitch glycolytic, while the light pink fibers were fast oxidative. All subcellular fractions of both fiber types had significant levels of CA activity; cytoplasmic and microsomal activity was significantly higher in light pink vs white fibers. Cytoplasmic CA from both fiber types was highly sensitive to the inhibitors acetazolamide and chlorzolamide, with Ki values of approximately 2 and 0.4 nM, respectively. Further analysis confirmed that cytoplasmic CA from both fiber types was kinetically similar to the high turnover Type II isoform. It appears that the evolution of the CA Type III isoform, found in vertebrate red muscle, did not occur with the differentiation of metabolic fiber types in crustaceans. Membrane-associated CA, which was also kinetically similar to the Type II isoform, was 20-fold higher in light pink fibers, suggesting a physiological role in facilitated CO2 efflux from the muscle fiber during periods of prolonged maximal activity. PMID- 11479904 TI - Negative test for cloacal drinking in a semi-aquatic turtle (Trachemys scripta), with comments on the functions of cloacal bursae. AB - Many aquatic turtles possess paired evaginations of the cloaca called cloacal bursae. Despite more than two centuries of study, little consensus exists as to the function(s) of these organs. We tested a recent suggestion that bursae could function in water uptake ("cloacal drinking"). Turtles (Trachemys scripta) were dehydrated (68-86% of maximum body mass) and given the opportunity to drink orally or cloacally. Dehydration caused increases in hematocrit and osmolality of extracellular fluid (ECF), but only after loss of 10-12% of maximum body mass, suggesting that turtles osmoregulated by reabsorbing water from the urinary bladder. Turtles drank eagerly when they could submerge their heads, and drinking was accompanied by an increase in body mass and a decrease in ECF osmolality. However, dehydrated turtles with tail and anus submerged showed no changes in mass or osmolality, suggesting that water absorption is not a significant function of the cloacal bursae in this species. Evidence for other putative functions is reviewed, leading to a pluralistic view: in cryptodires, bursae apparently function primarily in buoyancy control and secondarily in ion transport and nesting, but several pleurodires have been shown recently to use them in aquatic respiration. PMID- 11479905 TI - Phylogenetic distribution of apolipoproteins A-I and E in vertebrates as determined by Western blot analysis. AB - A putative apolipoprotein E (apoE) has been identified in the HDL and VHDL fractions of the turtle. This observation is of particular interest considering apoE has been reported absent in the domestic hen (Hermier et al., '95; Biochim Biophys Acta: 105-118, 1995) and thus presumed absent in nonmammalian vertebrates altogether. As a result, partial amino acid sequencing of this protein was performed and revealed that one fragment shared 41% sequence identity to human apoE. Western blot analysis using antisera to apoE demonstrated cross-reactivity to a 34-kDa protein (putative apoE) in turtle plasma. Further investigation using anti-apoE antibody in Western blot analysis detected immunoreactive apoE in the plasma of lamprey, spiny dogfish, skate, and alligator, but not in flounder, newt or python; its absence in several species of birds was confirmed. Using anti-apoA I antibody, apoA-I was detected in all vertebrate groups except a representative teleost (flounder). Apo-A-I antibody cross-reacted weakly with some putative apoE proteins (chicken, spiny dogfish and skate) and the reverse was true for anti apoE, which cross-reacted with putative apoA-I in birds, reptiles, and elasmobranchs, confirming the molecular similarity and phylogenetic relatedness of these two proteins. PMID- 11479906 TI - Bafilomycin A1 inhibits proteolytic cleavage and hydration but not yolk crystal disassembly or meiosis during maturation of sea bass oocytes. AB - Oocytes of the black sea bass, Centropristes striata, were enlarged in volume more than three-fold over a 24-hr period during oocyte maturation, both in vivo and in vitro. At the same time, the opaque oocytes clarified while the crystalline yolk inclusions lost their ordered structure, fused with one another, and formed a continuous electron-lucent mass. The oocyte size increase was due almost entirely to water uptake, which was accompanied by the accumulation of Na+, K+, and free amino acids (FAAs). The absolute amounts of each of these small molecular weight osmotic effectors increased 2x, 4x, and over 10x, respectively, indicating that the generation of FAAs is the major cause of water uptake during maturation. Amino acid analyses indicated that the amounts of all amino acids except taurine increased, so that selective amino acids were not produced during maturation. The increase in FAAs was accompanied by the loss of certain high molecular-weight yolk proteins and the generation of many smaller peptides. Oocytes stimulated to undergo maturation in the presence of bafilomycin A1, a specific inhibitor of the vacuolar ATPase-dependent proton pump, clarified and underwent maturation but did not increase significantly in size. Cytological examination revealed that yolk crystals fused and became homogeneous but maintained their electron density. No evidence of proteolysis was found in bafilomycin A1-treated oocytes and the generation of FAAs together with hydration was inhibited in a dose-dependent manner (I50 = 3 nM bafilomycin A1). Taken together, we postulate that the pronounced oocyte hydration in marine teleosts that spawn pelagic (floating) eggs is accomplished by a two-step process whereby (i) K+ influx promotes yolk crystal disassembly and yolk sphere fusion and (ii) acidification of the yolk spheres activates yolk proteolysis and concomitant hydration. Bafilomycin A1 inhibits only the second step so that many of the events of oocyte maturation, including germinal vesicle breakdown, occur in its presence but oocyte hydration is suppressed. PMID- 11479907 TI - Isolation and purification of the IGF-I protein complex from rabbit seminal plasma: Effects on sperm motility and viability. AB - A protein of about 150 kDa affecting sperm kinetic motility and viability was purified from rabbit seminal plasma. The incubation of rabbit sperm with this purified seminal plasma protein caused significant changes in sperm viability and motility. Moreover, the seminal protein showed a noticeable reactivating effect on immotile spermatozoa. A 10-mg amount of purified protein, added to immotile rabbit spermatozoa suspended in Tris-citrate, pH 7.4, resulted in a 48% reactivation. It is known that circulating insulin-like growth factors are bound to specific high-affinity binding proteins and form complexes with relative molecular masses of about 150 kDa. Western blotting analyses proved the existence of insulin-like growth factor in the protein purified from rabbit seminal plasma and immunofluorescence staining showed the existence of IGF-1 receptor in rabbit spermatozoa. Therefore, we suggest that the purified rabbit seminal plasma protein may represent the protein complex delivering IGF to the sperm cells thus affecting their physiological functions. PMID- 11479908 TI - Utilisation of nutrients by embryos of the enigmatic Australian viviparous skink Niveoscincus coventryi. AB - The Eugongylus species group of Australian lygosomine skinks provides an unparalleled opportunity to study the evolution of placentotrophy. Viviparity and placentotrophy have evolved in two lineages, currently recognised as the genera Pseudemoia and Niveoscincus. The genus Niveoscincus is important because it is the only lineage of squamates in which variation in placental morphology and in the pattern of embryonic nutrition is known. Niveoscincus coventryi has the least complex placental morphology among species currently assigned to the genus. We quantified the net uptake of nutrients across the placenta of N. coventryi for comparison with other species in the genus and with other viviparous and oviparous lizards. The pattern of embryonic nutrition of N. coventryi is similar to other viviparous lizards with simple placentae in that there is no net uptake of dry matter during development but there is a net uptake of water, calcium, potassium, and sodium. There is no net uptake of lipid, nitrogen (an index of protein), or magnesium. We conclude that N. coventryi is predominantly lecithotrophic. Further, if N. coventryi is the sister taxon to Tasmanian Niveoscincus, then the distribution of patterns of embryonic nutrition among members of this clade suggests that the evolution of placentotrophy occurred during radiation of this lineage in Tasmania. PMID- 11479909 TI - Differential scanning calorimetry studies of intraembryonic freezing and cryoprotectant penetration in zebrafish (Danio rerio) embryos. AB - Nucleation temperatures of intraembryonic water and cryoprotectant penetration in zebrafish embryos were studied using differential scanning calorimetry. The effects of embryo developmental stage, dechorionation, partial removal of yolk, cooling rate, and cryoprotectant treatment on the temperatures of intraembryonic freezing were investigated. Embryo stages were found to have a significant effect on the nucleation temperatures of intact embryos. Freeze onset temperatures of 11.9 +/- 1.5, -15.6 +/- 0.3, and -20.5 +/- 0.1 degrees C were obtained for intact embryos at 6-somite, prim-6, and high-pec stages, respectively. After dechorionation, the freeze onset temperatures of intraembryonic water shifted to significantly lower temperatures, being -23.5 +/- 0.8, -18.7 +/- 0.7, -24.9 +/- 0.8 degrees C for 6-somite, prim-6, and high-pec stages, respectively. Yolk reduced high-pec stage embryos showed significantly lower nucleation temperatures with an average onset at -27.9 +/- 0.4 degrees C. The effect of cryoprotectant treatment on the nucleation temperatures of intraembryonic water varies among different embryo stages and different cryoprotectants. Thirty-minute treatment with 2 M methanol significantly decreased the nucleation temperatures of dechorionated 6-somite embryos whilst no temperature decrease was observed for prim-6 or yolk-reduced high-pec embryos. Thirty-minute exposure to 1 M propylene glycol did not significantly affect the nucleation temperatures of dechorionated 6-somite, prim-6, or yolk-reduced high-pec embryos. In order to increase the permeability of embryos to cryoprotectants, the yolk sacs of dechorionated embryos at 6-somite or prim-6 embryos were punctured with a sharp micro-needle before exposure to cryoprotectants. The punctured prim-6 embryos showed significantly lower temperatures of intraembryonic freezing after 30 min of exposure to 2 M methanol following the multi-punctures. The nucleation temperatures of punctured 6-somite or prim-6 embryos were also decreased significantly after exposure to 1 M propylene glycol for 30 min. These results suggested that in intact embryos, intraembryonic freezing appeared to be seeded by the external ice in the perivitelline fluid and that in dechorionated embryos (in the absence of external water) intraembryonic freezing was more likely a consequence of heterogeneous nucleation. Methanol was demonstrated to show a limited degree of penetration into prim-6 stage embryos, but it did not penetrate later-stage embryos such as prim-6 and yolk-reduced high-pec. No propylene glycol permeation was observed for embryos at all stages. However, multi-punctures of yolk resulted in the permeation of both cryoprotectants into prim-6 embryos and propylene glycol permeation into 6-somite embryos. These findings may have important implications in overcoming the problem associated with the low membrane permeability of zebrafish embryos to cryoprotectants. PMID- 11479910 TI - Effects of the purified IGF-I complex on the capacitation and acrosome reaction of rabbit spermatozoa. AB - A protein complex containing IGF-I, purified from rabbit seminal plasma, was used to investigate its effects on the capacitation and acrosome reaction of rabbit spermatozoa. Uncapacitated sperm (Pattern F), capacitated sperm (Pattern B), and acrosome-reacted sperm (Pattern AR) were determined by CTC staining, and the results were validated by PSA-staining. The addition of the IGF-I complex to the capacitative medium directed the spermatozoa to spontaneous acrosome reaction. On the other hand, IGF-I complex, added to capacitated spermatozoa, acted as inducer of the acrosome reaction. Results of IVF experiments showed high rates of fertilization with capacitated spermatozoa, acrosome-reacted by either A23187 or IGF I complex, whereas significantly lower rates were obtained with spermatozoa capacitated in the presence of IGF-I complex. PMID- 11479911 TI - Structure of a character and the evolution of butterfly eyespot patterns. AB - All butterfly eyespots appear to be formed by a similar developmental mechanism. Thus the great diversity of patterns observed across different species of butterfly has evolved through changes in the genetic modulation of this mechanism. Our descriptions of both the development of eyespots and the genetic variation for eyespot traits in an African butterfly, Bicyclus anynana, are used here to discuss the most appropriate ways of applying the character concept to the evolution of butterfly eyespot patterns. J. Exp. Zool. (Mol. Dev. Evol.) 291:93-104, 2001. PMID- 11479912 TI - Origins of flower morphology. AB - Flowers evolved in many steps, probably starting long before flowering plants (angiophytes) originated. Certain parts of flowers are conservative and have not changed much during evolution; others are evolutionarily highly plastic. Here conservative features are discussed and an attempt is made to trace them back through their evolutionary history. Microsporangia and ovules (which develop into seeds) are preangiophyte floral elements. Angiospermy, combined with postgenital fusion, was the most prominent key innovation in angiophytes. Angiospermy and thecal organization of stamens originated earlier than all clades of extant angiosperms (the crown group of angiophytes). Differentiation of a perianth into calyx and corolla and syncarpy appeared after the first branching of the basalmost clades of extant angiosperms. Sympetaly and floral tubes as well as tenuinucellar, unitegmic ovules originated as major innovations in the clade that led to asterids. An obvious trend in flower evolution is increased synorganisation of parts, which led to new structures. Fixation of floral organ number and position was a precondition for synorganization. Concomitantly, plasticity changed from number and position of organs to shape of the new structures. Character distribution mapped onto cladograms indicates that key innovations do not appear suddenly, but start with trials and only later become deeply rooted genetically in the organization. This is implied from the common occurrence of reversals in the early history of an innovation. J. Exp. Zool. (Mol. Dev. Evol.) 291:105-115, 2001. PMID- 11479913 TI - Varieties of modules: kinds, levels, origins, and behaviors. AB - This article began as a review of a conference, organized by Gerhard Schlosser, entitled "Modularity in Development and Evolution." The conference was held at, and sponsored by, the Hanse Wissenschaftskolleg in Delmenhorst, Germany in May, 2000. The article subsequently metamorphosed into a literature and concept review as well as an analysis of the differences in current perspectives on modularity. Consequently, I refer to general aspects of the conference but do not review particular presentations. I divide modules into three kinds: structural, developmental, and physiological. Every module fulfills none, one, or multiple functional roles. Two further orthogonal distinctions are important in this context: module-kinds versus module-variants-of-a-kind and reproducer versus nonreproducer modules. I review criteria for individuation of modules and mechanisms for the phylogenetic origin of modularity. I discuss conceptual and methodological differences between developmental and evolutionary biologists, in particular the difference between integration and competition perspectives on individualization and modular behavior. The variety in views regarding modularity presents challenges that require resolution in order to attain a comprehensive, rather than a piecemeal and fragmentary, evolutionary developmental biology. J. Exp. Zool. (Mol. Dev. Evol.) 291:116-129, 2001. PMID- 11479914 TI - Evolutionary challenges of extreme environments (Part 2). PMID- 11479915 TI - Current problems with the zootype and the early evolution of Hox genes. AB - "Hox cluster type" genes have sparked intriguing attempts to unite all metazoan animals by a shared pattern of expression and genomic organization of a specific set of regulatory genes. The basic idea, the zootype concept, claims the conservation of a specific set of "Hox cluster type genes" in all metazoan animals, i.e., in the basal diploblasts as well as in the derived triploblastic animals. Depending on the data used and the type of analysis performed, different opposing views have been taken on this idea. We review here the sum of data currently available in a total evidence analysis, which includes morphological and the most recent molecular data. This analysis highlights several problems with the idea of a simple "Hox cluster type" synapomorphy between the diploblastic and triploblastic animals and suggests that the "zootype differentiation" of the Hox cluster most likely is an invention of the triploblasts. The view presented is compatible with the idea that early Hox gene evolution started with a single proto-Hox (possibly a paraHox) gene. J. Exp. Zool. (Mol. Dev. Evol.) 291:169-174, 2001. PMID- 11479916 TI - Macro-evolution of the hairy enhancer in Drosophila species. AB - It has been suggested that many of the changes in the developmental program might be in the cis-acting promoters and enhancer regions. Here I study the macro evolutionary changes of an enhancer region for the early developmental gene hairy in Drosophila melanogaster, D. simulans, D. pseudoobscura, D. willistoni, D. nebulosa, D. hydei, and D. virilis. The enhancer region is characterized by small, highly conserved blocks interspersed among highly variable regions. Nevertheless, species phylogenies constructed by the enhancer sequences agree with the widely accepted phylogeny of these species. The evolution of the variable regions is consistent with a molecular clock, while the evolution of the conserved blocks is significantly different from a clock. In particular, the D. pseudoobscura lineage shows the highest degree of species-specific change consistent with changes in expression timing reported in an earlier study. It has been suggested that the variation in sequence length between highly conserved blocks may play a role in the coordination of regulatory processes, such as protein-protein interactions; thus, stabilizing selection has been suggested to act on the length variations. Here I develop a test for stabilizing selection on length variation and show that the hairy enhancer does not show statistically significant evidence for stabilizing selection. J. Exp. Zool. (Mol. Dev. Evol.) 291:175-185, 2001. PMID- 11479917 TI - Role of motility in embryonic development I: Embryo movements and amnion contractions in the chick and the influence of illumination. AB - This study provides a quantitative analysis of the active movements of the chick embryo and of the contractions of the amnion over the entire developmental period of 21 days. Four types of embryo movements are distinguished. The motor activity of the embryo shows two characteristic peaks, with maximum contraction frequencies on the 12th and on the 16th day. In contrast, the amnion activity is higher at earlier stages and decreases as the body activity increases. The amnion activity is largely independent of the body activity. Illumination has a strong influence on embryo movements. It is shown that increases of light intensity affect the patterns of activity of both the embryo and the amnion. While the effect of light on the embryo can be interpreted as being transmitted via the optic system, the mechanism of the amniotic response is unclear. The results suggest that the amnion itself may be sensitive to light. J. Exp. Zool. (Mol. Dev. Evol.) 291:186-194, 2001. PMID- 11479918 TI - Testing the vulnerability of the phylotypic stage: on modularity and evolutionary conservation. AB - The phylotypic stage is the developmental stage at which vertebrates most resemble each other. In this study we test the plausibility of the hypotheses of Sander [1983, Development and Evolution, Cambridge University Press] and Raff [1994, Early Life on Earth, Columbia University Press; 1996, The Shape of Life, University of Chicago Press] that the phylotypic stage is conserved due to the intense and global interactivity occurring during that stage. First, we test the prediction that the phylotypic stage is much more vulnerable than any other stage. A search of the teratological literature shows that disturbances at this stage lead to a much higher mortality than in other stages, in accordance with the prediction. Second, we test whether that vulnerability is indeed caused by the interactiveness and lack of modularity of the inductions or, alternatively, is caused by some particularly vulnerable process going on at that time. From the pattern of multiple induced anomalies we conclude that it is indeed the interactiveness that is the root cause of the vulnerability. Together these results support the hypotheses of Sander and Raff. We end by presenting an argument on why the absence of modularity in the inductive interactions may also be the root cause of the conservation of the much discussed temporal and spatial colinearity of the Hox genes. J. Exp. Zool. (Mol. Dev. Evol.) 291:195-204, 2001. PMID- 11479919 TI - Monotreme IGF2 expression and ancestral origin of genomic imprinting. AB - IGF2 (insulin-like growth factor 2) and M6P/IGF2R (mannose 6-phosphate/insulin like growth factor 2 receptor) are imprinted in marsupials and eutherians but not in birds. These results along with the absence of M6P/IGF2R imprinting in the egg laying monotremes indicate that the parental imprinting of fetal growth regulatory genes may be unique to viviparous mammals. In this investigation, we have cloned IGF2 from two monotreme mammals, the platypus and echidna, to further investigate the origin of imprinting. We report herein that like M6P/IGF2R, IGF2 is not imprinted in monotremes. Thus, although IGF2 encodes for a highly conserved growth factor in chordates, it is only imprinted in therian mammals. These findings support a concurrent origin of IGF2 and M6P/IGF2R imprinting in the late Jurassic/early Cretaceous period. The absence of imprinting in monotremes, despite apparent interparental conflicts over maternal-offspring exchange, argues that a fortuitous congruency of genetic and epigenetic events may have limited the phylogenetic breadth of genomic imprinting to therian mammals. J. Exp. Zool. (Mol. Dev. Evol.) 291:205-212, 2001. PMID- 11479920 TI - 2-methoxyestradiol blocks cell-cycle progression at G(2)/M phase and inhibits growth of human prostate cancer cells. AB - 2-Methoxyestradiol (2-ME), an endogenous metabolite of 17beta-estradiol, is present in human blood and urine. Here we show for the first time that 2-ME significantly inhibited the growth of normal prostate epithelial cells and androgen-dependent LNCaP and androgen-independent DU145 prostate cancer cells. This growth inhibition was accompanied by a twofold increase in the G(2)/M population, with a concomitant decrease in the G(1) population, as shown by cell cycle analysis. 2-ME treatment affected the cell-cycle progression of prostate cancer cells specifically by blocking cells in the G(2) phase. Immunoblot analysis of the key cell-cycle regulatory proteins in the G(2)/M phase showed a 14-fold increase in the expression of p21 and an eightfold increase in the expression of p34 cell division cycle 2 (cdc2). We also found an accumulation of phosphorylated cdc2 after 2-ME treatment. Furthermore, Wee 1 kinase was detectable after 2-ME treatment. 2-ME treatment also led to an increase in the activity of caspase-3, followed by apoptosis, as shown by terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate-biotin nick end labeling and fluorescein isothiocyanate-poly(ADP-ribose) polymerase assay. Estrogen receptor levels did not change after treatment with 2-ME. Examination of the signaling pathways that mediate 2-ME-induced apoptosis showed reduction in the level of p53 expression and its DNA-binding activity. Given the fact that p53 mutations are common in patients with metastatic prostate cancer, our finding that 2-ME-mediated growth inhibition of human prostate cancer cells occurred in a p53-independent manner has considerable clinical significance. These findings, combined with the limited toxicity of 2-ME, may have significant implications for alternative treatment of advanced prostate cancer. PMID- 11479921 TI - Liver carcinogen aflatoxin B1 as an inducer of mitotic recombination in a human cell line. AB - The mycotoxin aflatoxin B1 (AFB1) is one of the most potent rodent and human liver carcinogens. Upon cytochrome P450-specific metabolism, it induces mutations as well as mitotic recombination events in in vitro systems. We have found that in the lower eukaryote yeast, the recombinagenic activity of AFB1 surpasses its mutagenic activity, and we speculated on possible consequences in terms of the mechanism of liver carcinogenesis. In this study we investigated whether the recombinagenic activity of AFB1 also would be identified in human cells. To address this question, we followed the fate of a heterozygous thymidine kinase (tk) allele in the human lymphoblastoid cell line TK6 upon exposure to AFB1. Individual mutants that had lost tk activity were subjected to loss of heterozygosity analysis of the tk locus and its flanking markers. Fluorescence in situ hybridization analysis on chromosome 17 also was performed. In parallel, a similar analysis was performed on TK6 cells exposed to the alkylating agent N nitrosomethylurea, a well-known classic point mutagen. Our analysis showed a difference in the molecular mechanism leading to inactivation of the tk allele upon exposure to these two mutagens. In AFB1-exposed cells the fraction of recombination-derived mutants predominated, whereas in N-nitrosomethylurea exposed cells the fraction of point mutants was higher. Thus, the recombinagenic activity of AFB1 previously identified in a lower eukaryote also was found in the human cell line TK6. Our data support the hypothesis that mitotic recombination represents a central mechanism of action in AFB1-induced liver carcinogenesis. PMID- 11479922 TI - Inositol hexaphosphate inhibits ultraviolet B-induced signal transduction. AB - Inositol hexaphosphate (InsP6) has an effective anticancer action in many experimental models in vivo and in vitro. Ultraviolet B (UVB) radiation is believed to be responsible for many of the carcinogenic effects related to sun exposure, and alteration in UVB-induced signal transduction is associated with UVB-induced carcinogenesis. Here we report the effects of InsP6 on UVB-induced signal transduction. InsP6 strongly blocked UVB-induced activator protein-1 (AP 1) and NF-kappaB transcriptional activities in a dose-dependent manner. InsP6 also suppressed UVB-induced AP-1 and nuclear factor kappaB (NF-kappaB) DNA binding activities and inhibited UVB-induced phosphorylation of extracellular signal-regulated protein kinases (Erks) and c-Jun NH2-terminal kinases (JNKs). Phosphorylation of p38 kinases was not affected. InsP6 also blocked UVB-induced phosphorylation of IkappaB-alpha, which is known to result in the inhibition of NF-kappaB transcriptional activity. InsP6 does not block UVB-induced phosphotidylinositol-3' (PI-3) kinase activity, suggesting that the inhibition of UVB-induced AP-1 and NF-kappaB activities by InsP6 is not mediated through PI-3 kinase. Because AP-1 and NF-kappaB are important nuclear transcription factors that are related to tumor promotion, our work suggests that InsP6 prevents UVB induced carcinogenesis by inhibiting AP-1 and NF-kappaB transcription activities. PMID- 11479923 TI - Loss of mismatch repair activity in simian virus 40 large T antigen-immortalized BPH-1 human prostatic epithelial cell line. AB - Simian virus 40 large T antigen (SVLTAg) has been used to immortalize cells; however, the mechanism leading to immortalization is still unclear. We hypothesize that DNA mismatch repair (MMR) activity is important during SVLTAg induced immortalization. To test this hypothesis, we used the SVLTAg-immortalized cell line BPH-1 derived from human benign prostate epithelial cells to analyze MMR activity and the expression of MMR genes (hMLH1, hPMS1, hPMS2, hMSH2, hMSH3, and hMSH6). The results demonstrated that BPH-1 cells were deficient in repairing G:T, A:C, and G:G mispairs in bacteriophage M13mp2. Reverse-transcription polymerase chain reaction experiments indicated MMR genes (hMSH3, hMSH6, and hPMS1) were expressed at a low level in BPH-1 cells. In contrast, all six MMR genes were expressed in human benign prostate hyperplasia tissues. Downregulation of hMSH3, hMSH6, and hPMS1 genes is not a result of the hypermethylation mechanism because demethylation with 5-aza-2'-deoxycytidine did not restore expression of these genes. Although the hMLH1 gene is expressed in BPH-1 cells, western blotting and exon analyses demonstrated that hMLH1 was mutated and/or deleted in BPH-1 cells. PMID- 11479924 TI - Inhibitory effect of glycolic acid on ultraviolet-induced skin tumorigenesis in SKH-1 hairless mice and its mechanism of action. AB - Glycolic acid, an alpha-hydroxy acid derived from fruit and milk sugars, has been used commonly as a cosmetic ingredient since it was discovered to have photoprotective and anti-inflammatory effects and antioxidant effects on ultraviolet (UV)B-irradiated skin. Little is known, however, about the functional role of glycolic acid on UV-induced skin tumorigenesis. In the present study, we examined the effect of glycolic acid on UV (UVA + UVB)-induced skin tumorigenesis and assessed several significant contributing factors in SKH-1 hairless mice. Inbred hairless female mice (15 animals/group) were irradiated for 5 d/wk at a total dose of 74.85 J/cm(2) UVA and 2.44 J/cm(2) UVB for 22 wk. Glycolic acid was applied topically twice a week at a dose of 8 mg/cm(2) immediately after UV irradiation. Glycolic acid reduced UV-induced skin tumor development. The protective effect of glycolic acid was a 20% reduction of skin tumor incidence, a 55% reduction of tumor multiplicity (average number of tumors/mouse), and a 47% decrease in the number of large tumors (larger than 2 mm). Glycolic acid also delayed the first appearance of tumor formation by about 3 wk. The inhibitory effect of glycolic acid on UV-induced tumor development was accompanied by decreased expression of the following UV-induced cell-cycle regulatory proteins: proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin E, and the associated subunits cyclin-dependent kinase 2 (cdk2) and cdk4. In addition, the expression of p38 kinase, jun N-terminal kinase (JNK), and mitogen-activated protein kinase kinase (MEK) also was lower in UV + glycolic acid-treated skin compared with expression in UV-irradiated skin. Moreover, transcription factors activator protein 1 (AP-1) and nuclear factor kappaB (NF-kappaB) activation was significantly lower in UV + glycolic acid-treated skin compared with activation in UV-irradiated skin. These results show that glycolic acid reduced UV-induced skin tumor development. The decreased expression of the cell-cycle regulatory proteins PCNA, cyclin D1, cyclin E, cdk2, and cdk4 and the signal mediators JNK, p38 kinase, and MEK may play a significant role in the inhibitory effect of glycolic acid on UV-induced skin tumor development. In addition, the inhibition of activation of transcription factors AP-1 and NF-kappaB could contribute significantly to the inhibitory effect of glycolic acid. PMID- 11479925 TI - A comprehensive analysis of loss of heterozygosity caused by hemizygous deletions in renal cell carcinoma using a subtraction library. AB - Several new loci were identified by a comprehensive analysis of loss of heterozygosity (LOH) using a subtraction library between matched normal and renal cell carcinoma (RCC) tissues. A total of 187 clones from the library, with a complexity of 1x10(4), were mapped, and 44 clusters of the mapped loci were subjected to LOH analysis using microsatellite markers. A total of 27 loci, which exhibited frequencies of LOH of at least 10% among 44 tumors, mostly clear-cell RCC, included several loci that were reported previously, such as, the von Hippel Lindau gene, adenomatous polyposis coli, and interferon regulatory factor-1, as well as new loci, at 5q32-q34, 6q21-q22, 8p12, and others. These loci exhibited LOH among 11.8-93.8% of tumors, and most, if not all, were derived from the sites of hemizygous deletions. The minimum regions of LOH of chromosomes 5, 6, and 8 were 9.0, 10.3, and 0.775 Mb, respectively. The average distance between the cloned fragments on the chromosomes was 2.2 Mb in 187 clones, indicating that the minimum LOH size expected from this subtraction analysis was roughly 50 kb. Therefore, the strategy described here provides comprehensive analysis of LOH sites, which were mostly caused by hemizygous deletions. PMID- 11479926 TI - Variant CJD epidemiology: joining up the dots. PMID- 11479927 TI - Silencing of gene expression: implications for design of retrovirus vectors. AB - Transcriptional silencing of retroviruses poses a major obstacle to their use as gene therapy vectors. Silencing is most pronounced in stem cells which are desirable targets for therapeutic gene delivery. Many vector designs combat silencing through cis-modifications of retroviral vector sequences. These designs include mutations of known retroviral silencer elements, addition of positive regulatory elements and insulator elements to protect the transgene from negative position effects. Similar strategies are being applied to lentiviral vectors that readily infect non-dividing quiescent stem cells. Collectively these cis modifications have significantly improved vector design but optimal expression may require additional intervention to escape completely the trans-factors that scan for foreign DNA, establish silencing in stem cells and maintain silencing in their progeny. Cytosine methylation of CpG sites was proposed to cause retroviral silencing over 20 years ago. However, several studies provide evidence that retrovirus silencing acts through methylase-independent mechanisms. We propose an alternative silencing mechanism initiated by a speculative stem cell-specific "somno-complex". Further understanding of retroviral silencing mechanisms will facilitate better gene therapy vector design and raise new strategies to block transcriptional silencing in transduced stem cells. PMID- 11479928 TI - Receptor for the group B coxsackieviruses and adenoviruses: CAR. AB - Considerable progress towards the characterisation of the long-sought receptor, CAR (coxsackievirus and adenovirus receptor), shared by group B coxsackieviruses (CVB) and most adenoviruses (Ad) has been made since it was isolated and cloned in 1997. The primary sequence of CAR shows that it is a member of the immunoglobulin superfamily of proteins, containing two Ig superfamily domains: an amino-terminal V-like module and a C2-like module. The CAR cytoplasmic domain, representing nearly one-third of the protein, is separated from the C2-like module by a single membrane-spanning sequence. The structure of the CAR V-like module complexed with the Ad fibre knob has been determined using recombinant proteins, and reveals three CAR modules associated with a single knob. Although recombinant CAR expressed in mammalian cells confers permissivity to CVB infection, details of the interaction between CAR and CVB remain to be elucidated. The expression of CAR appears to be highly regulated with respect to both cell type and developmental age. In rodents, CAR is expressed at high levels just before birth, and declines thereafter. Expressed levels have been found to increase in regenerating muscle and in response to immunological mediators or inflammation, and in RD cells and umbilical vein endothelial cells in response to high cell density. These studies indicate that CAR expression is highly regulated, but the mechanisms and molecules that mediate the expression remain to be discovered. The physiological function of CAR and its natural ligand also remain to be discovered. In addition, while CAR expression generally correlates with viral tropism, the relationship between the physiological function of CAR and the pathologies of CVB and Ad infections remain to be described. PMID- 11479929 TI - The pathogenesis of influenza in humans. AB - The rapid evolution of influenza A and B viruses contributes to annual influenza epidemics in humans. In addition, pandemics of influenza are also caused by influenza A viruses, whereas influenza B does not have the potential to cause pandemics because there is no animal reservoir of the virus. Study of the genetic differences between influenza A and influenza B viruses, which are restricted to humans, may be informative in understanding the factors that govern mammalian adaptation of influenza A viruses. Aquatic birds provide the natural reservoir for influenza A viruses, but in general, avian influenza is asymptomatic in feral birds. Occasionally, however, highly pathogenic strains of influenza cause serious systemic infections in domestic poultry. The pathogenicity of these strains is related to the presence of a polybasic cleavage sequence in the precursor of the surface glycoprotein haemagglutinin, which makes the glycoprotein susceptible to activation by ubiquitous proteases such as furin and PC6. However, the mechanism of pathogenicity may differ in highly pathogenic strains of human influenza, such as the H1N1 pandemic strain of 1918 and the H5N1 strain involved in the outbreak in Hong Kong in 1997. Binding of host proteases by the viral neuraminidase to assist activation of the haemagglutinin, shortening of the neuraminidase and substitutions in the polymerase gene, PB2, have all been suggested as alternative molecular correlates of pathogenicity of human influenza viruses. Additionally, systemic spread in humans of pathogenic subtypes has not been demonstrated and host factors such as interferons may be crucial in preventing the spread of the virus outside the respiratory tract. PMID- 11479930 TI - "Norwalk-like viruses" as a cause of foodborne disease outbreaks. AB - While outbreaks of foodborne disease remain an important public health concern, their aetiology is not identified in a majority of instances. In targeted studies, the application of newly developed molecular assays has demonstrated that a large proportion of these outbreaks may be caused by the "Norwalk-like viruses" (NLV), a genus of genetically related viruses belonging to the family Caliciviridae. NLV outbreaks associated with consumption of faecally contaminated oysters are frequently reported and can best be controlled by preventing contamination of oyster-harvesting waters. Infectious foodhandlers are another frequent source of contamination, and such transmission can be minimised by exclusion of ill foodhandlers and the maintenance of strict personal hygiene. Molecular assays have greatly refined the epidemiological investigation of foodborne NLV outbreaks, allowing the linking of outbreaks in different locations and permitting the identification of the virus in the implicated vehicle. The development of simpler and more sensitive assays and their use on a broader scale will assist in defining the true burden of foodborne NLV outbreaks and improve strategies for their prevention and control. PMID- 11479931 TI - Hepatitis B and hepatitis C co-infection in patients with HIV. AB - HAART has increased the life expectancy of patients with HIV. However, as their life expectancy increases, it becomes increasingly important to focus on the management of concurrent illnesses such as chronic HBV and HCV infections which have the potential to increase mid to long term morbidity and mortality. Shared epidemiological risks have resulted in the HIV infected population having a higher incidence of both HBV and HCV than those uninfected with HIV. Co-infection with HIV modifies the natural history of HBV infection, increasing the rate of viral replication, risk of carriage and chronic hepatitis but without increasing liver necroinflammatory processes. In chronic HCV infection, the presence of HIV enhances the risk of severe liver disease. There is no evidence as yet that HBV directly impacts on HIV disease progression but HCV infection increases the risk of death or an AIDS defining illness and impairs CD4+ T cell recovery during antiretroviral therapy. Treatment of either hepatitis virus is complex because of pharmacokinetic interactions with components of HAART regimens. PMID- 11479932 TI - Comparison of protein structures reveals monophyletic origin of the AdoMet dependent methyltransferase family and mechanistic convergence rather than recent differentiation of N4-cytosine and N6-adenine DNA methylation. AB - Phylogenetic analysis of the S-adenosyl-L-methionine-dependent methyltransferases was performed based on similarity of positions of main chain alpha-carbon atoms in published structures of members of this superfamily. The evolutionary tree was inferred and the problem of mono/polyphyletic origin of DNA methyltransferases from the Rossmann-fold enzymes was solved, bridging two seemingly antithetical hypotheses. The comparison of protein structures provides evidence for an evolutionary link between widely diverged subfamilies of RNA and DNA N6-adenine methyltransferases and argues against the close homology of N6-adenine and N4 cytosine methyltransferases, apparent from biochemical data and comparison of fragments of sequences. Such evolutionary analysis of methyltransferases has never been published yet in the literature and will guide further phylogenetical studies based on both sequence and structure comparison. PMID- 11479933 TI - Curved DNA in promoter sequences. AB - Sequence-dependent DNA curvature is known to play an important role in initiation of transcription of many genes. We compared the distribution of predicted intrinsic curvature of Escherichia coli and human promoter sequences with the distribution of curvature of randomly selected coding and non-coding fragments from these organisms. Different methods of curvature calculation were found to yield mostly similar overall tendencies of DNA curvature in all groups of sequences. According to all methods of calculation, E. coli promoters were found to be more curved than coding sequences from the same genome and random sequences with the same nucleotide composition. By contrast, the average curvature of human promoter sequences was only marginally greater than the average curvature of human coding sequences. Non-coding intron sequences were found to be the most curved of the human sequences examined. Based on these observations, we hypothesize about the role of DNA curvature in promoter sequences. PMID- 11479934 TI - Comparative analysis of the Arabidopsis and rice expressed sequence tag (EST) sets. AB - Large numbers of expressed sequence tags (ESTs) have now been generated from a variety of model organisms. In plants, substantial collections of ESTs are available for Arabidopsis and rice, in each case representing significant proportions of the estimated total numbers of genes. Large-scale comparisons of Arabidopsis and rice sequences are especially interesting due to the fact that these two species are representatives of the two subclasses of the flowering plants (Dicotyledonae and Monocotyledonae, respectively). Here we present the results of systematic analysis of the Arabidopsis and rice EST sets. Non redundant sets of sequences from Arabidopsis and rice were first separately derived and then combined so that gene families in common between the two species could be identified. Our results show that 58% of non-singleton ESTs are derived from genes in gene families common to the two species. These gene families constitute the basis of a core set of higher plant genes. PMID- 11479935 TI - Pathophysiology of calcium deposition in coronary arteries. AB - BACKGROUND AND MORPHOLOGIC STUDIES: Because coronary artery calcification correlates highly with plaque burden, it is an excellent disease marker for atherosclerosis. However, it is not a sensitive indicator of disease activity, and does not predict luminal compromise because of compensatory remodeling. In addition, most data do not support the concept that plaque calcification is related to plaque instability. Plaques demonstrating acute rupture usually show mild or moderate calcification, and biophysical models do not predict that calcium should result in an increased propensity to rupture. This review outlines morphologic studies relating calcification to risk factors and coronary plaque morphology. PMID- 11479936 TI - Biology of calcification in vascular cells: intima versus media. AB - BACKGROUND: Vascular calcification occurs at two distinct sites within the vessel wall: the intima and the media. Intimal calcification occurs in the context of atherosclerosis, associated with lipid, macrophages and vascular smooth muscle cells, whereas medial calcification can exist independently of atherosclerosis and is associated with elastin and vascular smooth muscle cells. PATHOGENESIS: In this review we compare intimal and medial calcification, particularly discussing the mechanisms which may be responsible for each type of calcification. Similar mechanisms probably initiate and regulate both forms of calcification including the generation of matrix vesicles/apoptotic bodies and local expression of mineralization-regulating proteins. However, since different modifying agents such as lipids in the intima and elastin in the media are present at the sites of calcification and are associated with particular diseases, this implies that the etiologies of these processes differ. For example, intimal calcification is associated with atherosclerosis while medial calcification occurs commonly in the diabetic neuropathic leg. CLINICAL IMPORTANCE: Since both types of calcification correlate with significant morbidity and mortality, we discuss the different types of calcification in terms of their clinical importance. PMID- 11479937 TI - Prognostic implications of absolute and relative calcium scores. AB - BACKGROUND: The continuing epidemic of cardiovascular disease underscores the need for more effective atherosclerosis detection and prevention. In fact, though the mortality rate decreased during the past decades, the morbidity rate continued to climb. The use of conventional risk factors is helpful to assess the median risk of a population, but it is unsatisfactory to estimate the actual risk of an individual patient. MODERN IMAGING TECHNOLOGIES: As a consequence, modern imaging technologies able to detect silent atherosclerotic disease have elicited a strong interest in the medical community with the hope that they may render risk stratification more accurate. Technologies such as electron beam tomography (EBT) are expected to bridge the gap between the mere presence of risk factors for disease development and assessment of the actual individual risk of events based on the visualization and quantification of silent disease. CONCLUSION: The medical literature is currently pervaded by a vivacious debate regarding the ability of EBT to provide such information as some investigators still hold concerns about the cost and effectiveness of a preventive approach driven by technology. PMID- 11479938 TI - [Importance of ECG-triggering with the multi-slice CT for ruling out coronary calcification in individuals without diagnosed coronary disease]. AB - BACKGROUND: To avoid unnecessary cardiac catheterization, ruling out coronary calcification has become increasingly important. On the other hand, the detection of coronary calcification is proof of coronary atherosclerosis and therefore requires a reduction of the LDL-cholesterol < 100 mg/ml according to the NCEP guidelines. The "gold standard" for calcium scanning is traditionally ECG triggered electron beam tomography (EBT). In addition, mechanical multi-slice CTs (MSCT) have been used without ECG-triggering for more than 6 years for calcium scanning. PATIENTS AND METHODS: The importance of ECG-triggering was assessed by examining apparently healthy subjects referred by their physicians (n = 1,206, 57 +/- 10 years, 75% male) using an Mx-8000 four-slice CT (Marconi, USA). Data were obtained from the MUNICH-Registry (MUltislice Normal Incidence of Coronary Health). Image acquisition without ECG-triggering was performed in spiral mode at 120 kV and 249 mAs using a collimation of 2.5 mm, resulting in an effective slice width of 3.2 mm. Prospective ECG-triggering was achieved in sequential ("axial") mode at 120 kV and 165 mAs using an effective collimation of 2.5 mm. The trigger was set at 333 ms prior to 90% of the expected RR-interval. RESULTS: With both groups well comparable regarding age, sex and risk factors, calcium was ruled out in 51.5% of the individuals without and in 34% of the persons with ECG-triggering (p < 0.001). An Agatston score > 100 was detected in 14.2% without and in 33.1% with ECG-trigger. The loge-transformed calcium scores were 1.7 +/- 2.2 for persons without and 2.9 +/- 2.6 with ECG-trigger (p < 0.001). Multivariate linear regression analysis showed that the application of ECG-triggering was an independent and the strongest predictor for the proof of calcium. With ECG triggering, calcium was detected in 1.51 +/- 1.42 major coronary arteries (median 1.0), whereas without ECG-triggering, calcium was detected in 1.03 +/- 1.33 major coronary arteries (median 0.0, p < 0.001). CONCLUSION: Since without ECG triggering every fifth patient with coronary atherosclerosis is erroneously classified as "healthy" and only the sensitive exclusion of coronary calcium is helpful in avoiding unnecessary cardiac catheterizations, the use of ECG triggering for calcium scanning with MSCT is mandatory. PMID- 11479939 TI - Recommendations for standardization of EBT and MSCT scanning. AB - BACKGROUND: The detection and quantification of coronary calcifications by electron beam tomography (EBT) permits to diagnose coronary atherosclerosis in an early stage. Initial reports indicate that multislice spiral CT (MSCT) also permits the quantification of coronary calcium, while equivalency to EBT has not been definitely proven. Since image acquisition, reconstruction and evaluation parameters influence the results of calcium quantification with CT techniques, standardization of the investigation is mandatory to make results comparable. AIM: The article summarizes guidelines for image acquisition and evaluation by EBT and describes guidelines and consensus reports that were issued concerning the clinical use of the method. PMID- 11479940 TI - [Progression of coronary calcium]. AB - BACKGROUND: The progression of coronary atherosclerosis is determined by risk factor exposure. It is also an important predictor of future cardiac events in its own right. Coronary calcium represents an integral part of coronary atherosclerosis. Accurate measurements can be obtained by using electron-beam computed tomography (EBT) or recently developed multidetector spiral CT scanners which are in the state of evaluation. The variability of EBT-derived quantification of coronary calcium is < 10% and thus suitable for determining the progression of coronary calcium. PROGRESSION OF CORONARY CALCIUM: In the healthy general population, the annual progression measures approximately 24%, whereas in untreated, asymptomatic high-risk patients, it is 40-50%. Similar values are observed in symptomatic patients with modern pharmacological therapy. There is a considerable amount of inter-individual variability in the progression of coronary calcium, probably as a result of the complex interplay of numerous factors which influence the process. Progression of coronary calcium is observed at typical predilection sites of atherosclerosis in the coronary tree. Changes in overall coronary calcium result from uniform changes at these sites. This uniform pattern of change suggests that the development of calcified plaque disease is a coronary systemic process. The baseline amount of coronary calcium is significantly associated with its progression. In patients with a high baseline calcium score, absolute progression is enhanced. LDL-cholesterol is the most important risk factor influencing progression. Indeed, the effect of medical therapy on coronary atherosclerosis can be assessed. Preliminary data indicate that clear-cut progression of coronary calcium is associated with an increased risk of cardiac events. CONCLUSION: For the first time, EBT-derived measurements of the progression of coronary calcium enable direct assessment of the activity of coronary calcified atherosclerosis. The interaction between therapeutic measures, progression of anatomical disease, and clinical course can be appreciated. PMID- 11479941 TI - Left ventricular hypertrabeculation in myotonic dystrophy type 1. AB - BACKGROUND: Left ventricular hypertrabeculation (LVHT) has not been described in myotonic dystrophy Type I (MD1) before. CASE REPORT: A 42-year-old man developed typical features of MD1 since 1992. Creatinekinase was slightly, but recurrently elevated. Needle electromyograms were myogenic and showed extensive spontaneous activity. Muscle biopsy was compatible with MDI. DNA analysis revealed a heterozygous 300 CTG-repeat expansion in the myotonic-dystrophy proteinkinase gene on chromosome 19q13.3. Cardiac history and clinical cardiologic examination were normal. On ECG, ST elevation and atrial flutter were found. The AECG was normal except for atrial flutter. Surprisingly, transthoracic echocardiography revealed LVHT, previously described only in Becker's muscular dystrophy, mitochondriopathies, and Barth syndrome. CONCLUSION: A rare cardiac manifestation of MD1 may be LVHT which alone has no therapeutic implication. PMID- 11479942 TI - [Ablation of atrial extrasystoles and atrial fibrillation]. PMID- 11479943 TI - [Clinical analysis of 12 cases with venous thromboembolism during pregnancy and puerperium]. AB - OBJECTIVE: To study the incidence, cause and clinical manifestation of venous thromboembolism during pregnancy and puerperium, and its diagnosis and treatment. METHODS: 12 cases of venous thromboembolism admitted in our hospital from 1984 1997 were analysed retrospectively. The plasma protein C, protein S and antithrombin III activities were measured in 4 of the cases and activated protein C resistance (APC-R) were assayed by activated partial thrombinplastin time (APIT) in the presence and absence of APC (APC-APIT) and FV Leiden gene mutation were analysed by PCR restriction fragment length polymorphisms (RFLP) methods as well. RESULTS: Four cases occurred before delivery and 8 postpartum. Two cases complicated by pulmonary thromboembolism, and of them 1 died. APC-R(+), reduced activity of antithrombin-III and protein S each were found in 3 separate cases. No FV Leiden gene mutation was found in the 4 cases. CONCLUSIONS: The formation of venous thromboembolism during pregnancy and puerperium is highly associated with the deficiency of anticoagulant proteins. Anticoagulation is recommended in the high risk women of thromboembolism. PMID- 11479944 TI - [Expression of apoptosis regulatory protein bcl-2 and bax in endometrium of polycystic ovary syndrome]. AB - OBJECTIVE: To study the expression and implication of apoptosis regulatory protein bcl-2 and bax in endometrium of polycystic ovary syndrome (PCOS). METHODS: The apoptosis regulatory protein expression of bcl-2 and bax protein in endometrium were investigated by immunohistochemical method in 18 PCOS and 27 normal subjects proliferative phase 10 and secretory phase 17. RESULTS: Bcl-2 expression was higher in PCOS than that both in control proliferative phase (P < 0.05) and secretory phase (P < 0.001). Bax expression was not significantly different between PCOS and controls (P > 0.05). CONCLUSIONS: The higher bcl-2 protein expression inhibits the endometrium apoptosis and plays an important role in the abnormal proliferative changes of endometrium in PCOS. PMID- 11479945 TI - [Analyses of the factors affecting the occurrence of polypronuclear embryos in in vitro fertilization and embryo transfer]. AB - OBJECTIVE: To investigate the factors affecting the occurrences of the polypronuclear embryos in in-vitro fertilization and embryo transfer (IVF-ET). METHODS: The data from 145 IVF-ET cycles and 1,168 oocytes were analyzed by logistic regression for multiple factors. The influences of the couple's age, the superovulation stimulation protocols, the number of the follicles punctured and the oocytes obtained, the preinseminational interval, the quality and quantity of seminal and inseminational sperms, and the oocyte fertilization ratio on the incidences of the polypronuclear embryos from the matured, and immature oocytes were examined respectively. RESULTS: The incidence of the polypronuclear embryos from matured oocytes was significantly negatively correlated with the women age and the number of the follicles punctured, and the incidence from immature oocytes was significantly positively correlated with the density of sperms at grade a and b and the oocyte fertilization ratio. CONCLUSIONS: The woman's age and the number of follicles are the main factors which influence the occurrence of polypronuclear embryos from matured oocytes, while the density of the seminal sperm with high motility and the fertilization ratio play important roles in the occurrence of polypronuclear embryos in immature oocytes. Stimulating greater number of follicles might be benefit for the reduction of occurrence of polypronuclear embryos. PMID- 11479946 TI - [Injury to urinary system in the operation of gynecology]. AB - OBJECTIVE: To reduce the incidence of urinary system injury from gynecology operations. METHODS: 26 cases with urinary system injuries during gynecological operations were analyzed from 1982 to 1996. RESULTS: The incidence of urinary injury in this hospital was 0.34%, and injuries occurred most frequently in the bladder. Among the 17 cases of bladder injuries, 12 occurred during tubal ligation, 3 during cesarean section, and 2 in myomectomy. There were 9 cases of ureter injuries, 6 cases occurred in radical hysterectomy for cervical cancer, 1 in cesarean section with hysterectomy, and 2 in hysterectomy with broad ligament leiomyoma. CONCLUSIONS: This study demonstrated that it was important to be familiar with the regional anatomy, to operate strictly and separate those organs carefully in preventing the urinary system injury. PMID- 11479947 TI - [In vitro study of 6B11GM to induce cellular immunity in patients with ovarian carcinoma]. AB - OBJECTIVE: In order to explore the possibility of ovarian carcinoma anti idiotypic single chain antibody (6B11scFV) and human granueocyte-macrophage colony stimulating factor (hGM-CSF) fusion protein 6B11GM as tumor vaccine. METHODS: Mononuclear cells, T cells and monocyte used as antigen presenting cell (APC) were separated from peripheral blood, and tumor cells were obtained from ascitic fluid in the same patients. In vitro T cell proliferation and autologous tumor cell cytotoxicity assay were applied. RESULTS: 6B11GM could induce T cell proliferation in vitro which was antigen specific. Cytotoxicity of mononuclear cells to autologous tumor cells increased after primed with 6B11GM. CONCLUSION: 6B11GM could induce efficient cellular immunity in patients with ovarian carcinoma, hopefully may be used for ovarian carcinoma vaccine. PMID- 11479948 TI - [Radioimmunodetection in the diagnosis of trophoblastic disease]. AB - OBJECTIVE: To study the efficiency of radioimmunodetection (RID) to locate the loci of trophoblastic disease. METHOD: 24 RIDs were performed with 131I labeled mouse anti-hCG monoclonal antibodies on 21 patients with trophoblastic disease. RESULTS: Of 24 RIDs performed, 13 were positive, including one false positive; 11 were negative, including two false negative. The sensitivity of RID was 85.7%, the specificity was 90.0%, the accuracy was 87.5%. In 8 patients who underwent operations after RID, the findings of RID were in accordance with the pathologic and immunohistologic findings. CONCLUSIONS: RID is an efficient imaging method for trophoblastic disease. In some cases, RID showed its value as a functional diagnostic method. PMID- 11479949 TI - [The relationship between angiogenesis and p53 overexpression in endometrioid adenocarcinoma]. AB - OBJECTIVE: To determine the relationship between angiogenesis and p53 overexpression and explore the relationship between angiogenesis and clinicopathologic features in endometrioid adenocarcinoma. METHODS: Angiogenesis was evaluated by intratumoral microvessel density (IMD). Eighty-five endometrioid adenocarcinomas' IMD were identified by immunohistochemical staining of endothelial cells for factor VIII-related antigen, and the expression of p53 protein was stained with the same method by it's monoclone antibody. RESULTS: (1) The positive rate of p53 expression in endometrioid adenocarcinoma was 50.6%. The positive rate in stage III-IV (69.2%) was higher than that in stage I-II(42.4%) (P < 0.05). (2) IMD was higher in stage III-IV (13.27 +/- 6.14) than that in stage I-II (8.31 +/- 4.78) (P < 0.01). IMD in G1, G2 and G3 was 8.95 +/- 5.51, 9.28 +/- 4.87 and 14.73 +/- 7.66, respectively (P < 0.05). In groups of non myoinvasion, < 1/2 myoinvasion and > or = 1/2 myoinvasion, IMD was 8.45 +/- 5.53, 8.95 +/- 4.99 and 11.96 +/- 6.39, respectively (P < 0.05). (3) In negative, weakly positive and strong positive groups of p53 overexpression, IMD was 5.52 +/ 2.11, 11.75 +/- 4.18 and 16.00 +/- 4.63, respectively (P < 0.01). CONCLUSIONS: Angiogenesis tightly correlated with clinicopathologic factors, such as stage, grade and myoinvasion, is promoted by p53 overexpression in endometrioid adenocarcinoma. It can be concluded that angiogenesis may act as a predictor of prognosis of endometrioid adenocarcinoma. PMID- 11479950 TI - [Effects of estradiol and medroxyprogesterone on the growth and doxorubin resistance of drug-resistant human epithelial ovarian cancer cell line OVCAR-3]. AB - OBJECTIVE: To study the effects of estradiol (E2) and medroxyprogesterone acetate (MPA) on the growth of drug resistant human epithelial ovarian cancer cell line. METHODS: OVCAR-3 cells, treated by different concentrations of E2, MPA and verapamil, were examined for their growth inhibitions with methyl thiazolyl tetrazolium (MTT, a tetrazolium dye) rapid photocolorimetric assay. RESULTS: OVCAR-3 cell growth was inhibited by E2 at high concentrations (10.00-100.00 mumol/L) and by MPA at 100.00 mumol/L, while lower concentrations of E2 (0.01 1.00 mumol/L) and MPA (0.01-10.00 mumol/L) had no effect on OVCAR-3 cell growth, neither stimulation nor inhibition. The effect of E2 combined with MPA was more obvious than either one used separately. E2 and MPA in combination with doxorubin showed a synergistic inhibition effect on OVCAR-3 cell growth, which suggested that both of them could reverse the doxorubin resistance of OVCAR-3 cell line. The inhibition effect of Verapamil combined with doxorubin was obviously weaker than that of E2 and MPA. CONCLUSIONS: E2 and MPA could reverse the doxorubin resistance of OVCAR-3 cell line and their inhibition were more obvious than that of verapamil, which suggested that E2 and MPA may be used clinically for adjuvant chemotherapy. PMID- 11479951 TI - [Clinical experience with 7 years of norplant]. AB - OBJECTIVE: To assess the efficacy, safety of Norplant implants used by Chinese women. METHOD: In a prospective, multicenter clinical trial, 10,718 women receiving Norplant implants were monitored annually through 5 years and part of women continuously used the implants up to 7 years. RESULTS: The five-year pregnancy rate was 0.63/100 users. 7,554 women completed 5 years, 3,622 women completed 6 years and 2,433 women completed 7 years. The annual pregnancy rates were 0.40/100 users in both year 6 and year 7. The pregnancy rates increased with weight (P < 0.05) and decreased with age. The menstrual disturbances were common, also the main reason for discontinuation. Headache, dizziness and weight gain led to termination more frequently than did other adverse events apart from menstrual disturbances. The 5-year continuation rate was 72.1/100 users. Among aged > 30 and weight < 60 kg, the implants may be used for 7 years. CONCLUSION: It's proved that the Norplant implants is a highly effective, long acting, safe, simple and reversible contraceptive method, and it is acceptable in Chinese women. PMID- 11479952 TI - [Insulin resistance in patients with polycystic ovary syndrome]. PMID- 11479953 TI - [Advances in the treatment of uterine sarcoma]. PMID- 11479954 TI - [The clinical study on urinary albumin and calcium output in 24 hours to serve as early markers for pregnancy induced hypertension]. AB - OBJECTIVE: To assess the efficacy of urinary calcium and albumin output in 24 hours for prediction of the occurrence of pregnancy induced hypertension (PIH). METHODS: 24-hour urine samples were collected at 24-32 weeks' gestation in 98 health women for the measurement of calcium and albumin, at the same time plasma calcium and creatinine were also determined. All patients were followed up until delivery and were classified subsequently according to the occurrence of PIH. Microalbumin was measured by RIA and calcium by automatic biochemistry analysis. RESULTS: The patients in whom PIH developed later (n = 14) had significant lower urinary calcium excretion [(1.43 +/- 0.37) mmol/24 h VS (3.26 +/- 0.75) mmol/24 h] and higher microalbuminuria [(12.68 +/- 6.81) micrograms/24 h VS (6.08 +/- 3.48) micrograms/24 h] as compared with the group which remained normal (n = 84). CONCLUSIONS: The decrease of calcium and increase of albumin per 24 hours of urine were observed 4-8 weeks before the onset of PIH. The above observation may serve as early markers for PIH. PMID- 11479955 TI - [Study on thrombomodulin and plasminogen activator inhibitor type-1 in patients of pregnancy induced hypertension]. AB - OBJECTIVE: To study the circulating levels of Thrombomodulin (TM) and Plasminogen activator inhibitor type-1 (PAI-1) in women with pregnancy induced hypertension (PIH) and normal pregnant women. METHODS: Blood samples were withdrawn from 73 pregnant women, In which 25 mild PIH, 26 moderate PIH, 22 severe PIH were divided into study group, and 24 normotensive healthy pregnant women in their third trimester were control group. The determination of TM was measured by ELISA and PAI-1 by Colorimetric Assay methods. RESULTS: Circulation levels of TM and PAI-1 were increased with increasing severity of the PIH. There were no significant differences between groups of mild PIH and normal pregnancy (TM 45.69 +/- 15.15 micrograms/L and 40.14 +/- 15.82 micrograms/L, PAI-1 22.06 +/- 12.13 kU/L and 18.40 +/- 9.78 kU/L respectively). The parameters significantly increased in moderate (TM 52.76 +/- 13.91 micrograms/L, PAI-1 25.77 +/- 9.45 kU/L) and severe PIH (TM 64.84 +/- 15.16 micrograms/L, PAI-1 30.79 +/- 11.64 kU/L) groups. CONCLUSIONS: These results demonstrate that the endothelial cell damage and its functional disturbance play a very important role in the etiology of PIH. TM and PAI-1 may serve as clinical markers for the assessment of endothelial damage in PIH. PMID- 11479956 TI - [Detection of antineutrophil cytoplasmic autoantibodies in pregnancy induced hypertension and its clinical significance]. AB - OBJECTIVE: To explore the level of antineutrophil cytoplasmic autoantibodies (ANCA) in patients with pregnancy induced hypertension (PIH) and its clinical significance. METHOD: 37 cases with PIH and 39 non-PIH pregnancy were studied for ANCA by indirect immunflourscence. RESULTS: ANCA positive rates were 33.3%, 88.9% and 86.4% in mild, moderate, severe PIH, respectively, and most of them were c ANCA. The positive rate was only 13.2% in non-PIH control, significantly lower than those in PIH (P < 0.001). CONCLUSION: ANCA may be one of the causes for endothelial cell damage and a predicting marker in PIH. PMID- 11479957 TI - [Changes of insulin and C-peptide in pregnancy of gestational diabetes mellitus]. AB - OBJECTIVE: To observe the different changes of plasma insulin and c-peptide in patients with gestational diabetes mellitus (GDM), gestational impaired glucose tolerance (GIGT), normal pregnant women, and to detect the function of beta-cell in patients with gestational impaired glucose metabolism. METHODS: Test the plasma insulin, c-peptide in 48 cases of GDM(group I), 39 cases of GIGT (group II), 39 cases of normal pregnant women (group III) and 22 cases of healthy non pregnant women (group IV) by radioimmunoassay. RESULTS: (1) In third trimester of group III, insulin, c-peptide levels were higher than those of group IV (P < 0.01). They increased from 28-34 weeks to 38-40 weeks, peaked during delivery. Insulin levels of group I and group II were higher than those of group III (P < 0.01, P < 0.05), but tended to decrease by the end of the third trimester and peaked during delivery. During delivery, there was no significant differences among the three groups. At 28-40 weeks, c-peptide of group I was significantly higher than those in groups II and III (P < 0.01) and was lower than that in group III during delivery (P < 0.05). Insulin and c-peptide of these 3 groups dropped gradually within 2 months after delivery but in group I and II they were significantly higher than those of group III (P < 0.01). CONCLUSION: In third trimester insulin and c-peptide of group I-III increased till delivery, the reason maybe associated with insulin resistance. GDM had the same peaking secretion during delivery with normal pregnant women and GIGT group, this maybe due to the stress during delivery. Insulin levels of most GDM and GIGT were still higher than normal after two month postpartum, and should be followed up carefully. PMID- 11479958 TI - [Study on the relationship between first ovulation and infant feeding in lactating postpartum women]. AB - OBJECTIVE: To determine the first ovulation after delivery and to provide scientific data for contraception in lactating postpartum women. METHODS: From January 1996 to December 1998, 101 lactating women were continuously monitored of their ovulation by ultrasound, basal body temperature (BBT) measurement and cervical mucus examination. RESULTS: The evidences of first ovulation (follicle > 1.8 cm in diameter) were found by ultrasound in 53 (52.5%) women during 154 days after delivery on the average and there were 11 (10.9%, 11/101) subjects who had first ovulation during 4 months after delivery and 42 (41.6%, 42/101) subjects had, after 4 months. Significant positive correlation was found between the time of supplementary feeding and the time of the first menses (n = 100, r = 0.4764, P < 0.01) and first ovulation (n = 53, r = 0.5554, P < 0.01). CONCLUSION: For postpartum mother it is suitable to begin with birth control method at the 4th month after delivery and to prolong the breast feeding is also a good method for birth control. PMID- 11479959 TI - [Clinical analysis of 50 cases of transvaginal hysterectomy]. AB - OBJECTIVE: To assess the advantage, indications, surgical management and the clinical value of transvaginal hysterectomy in patients without uterine prolapse. METHODS: From Oct. 1995 to Nov. 1998, 50 women undergone transvaginal hysterectomy for various benign gynecological lesions were consecutively included in this study. Their mean age was 45 years (range 36-50). There were 10 patients (20%) with normal uterine size, and 16 (32%) with uteri of > 12 gestational weeksize (range 14-16 weeks) and the remaining 24(48%) < or = 12 weeksize. 11 patients (22%) underwent salpingo-oophorectomy simultaneously, 6 patients (12%) had previous laparotomy. RESULTS: The mean operating time was 63 min with a range 40-110 min. Post-operative hospital stay averaged 4 days (range 2-8 days). The amount of blood loss during operation ranged 50-100 ml. The only postoperative complication was small residue hematoma in 1 case and cured with conservative treatment. CONCLUSIONS: Transvaginal hysterectomy was associated with remarkable advantages: shorter operating time and hospital stay, fewer complication, less discomfort, quicker recovery and less expenses. Enlarged uterus to a size of > 12 gestational weeks; benign adnexal mass or previous operation should no longer be considered as contraindication of transvaginal hysterectomy. PMID- 11479960 TI - [Effects of synthetic peptides on ovarian cancer cells]. AB - OBJECTIVE: To observe the effect of follicle-stimulating hormone(FSH) synthetic peptides(FSH binding Fragment), FSH and synthetic peptides on the proliferation of human epithelial ovarian cancer cell. METHODS: Human epithelial ovarian cancer cells lines SKOV3, OVCAR, AO and 3AO were incubated with FSH, FSH binding fragment, FSH and the binding fragment respectively. The cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) technique. RESULTS: The rate of proliferation in the cancer cell was increased apparently as increasing in the concentration of FSH and the rate of proliferation average value 28.0%, and was decreased apparently as increasing in the concentration of the synthetic peptides and the rate of inhibition average value 8.3%. When the cell was expressed in FSH, the proliferation was decreased apparently as increasing in the concentration of the peptides and the rate of inhibition average valve 3.1%. CONCLUSIONS: It is suggested that FSH binding fragment can inhibit the proliferation of ovarian cancer cell. The FSH binding fragment could be used as a binding part of anticancerous complex for ovarian cancer. PMID- 11479961 TI - [The association of radiation-induced apoptosis with radiosensitivity of cervical carcinoma]. AB - OBJECTIVE: To study the relationship between apoptosis, radiation-induced apoptosis and radiosensitivity of cervical carcinoma and find new markers for predicting radiosensitivity. METHODS: TdT-mediated bio-dUTP nick end labeling (TUNEL) method was used to check apoptosis cells. The association of radiation induced apoptosis with tumor volume shrinkage, radiation damage grade and local control rate was analysed. RESULTS: There was no relationship between apoptosis before radiation and tumor volume shrinkage(P > 0.05); Radiation-induced apoptosis had a significant association with tumor volume shrinkage (P < 0.01); In patients with radiation-induced apoptosis 10 or more, the grade of radiation damage IIb-III accounted for 76% (13/17), while for less than 10, it was 39% (5/13, P < 0.05); After 6 months of radiotherapy, local control rate was 88% (15/17) in the former, 69% (9/13) in the latter, respectively (P > 0.05). CONCLUSION: Radiation-induced apoptosis has some reference value for predicting radiosensitivity of cervical carcinoma. PMID- 11479962 TI - [Female malignant tumors of genital organs with pericardial effusion: report of 9 cases and review of literatures]. AB - OBJECTIVE: To study the clinical characteristics and the management of malignant tumors of genital organs with pericardial effusion. METHODS: Nine cases of gynecologic malignant tumors with pericardial effusion were analyzed retrospectively. RESULTS: Nine cases of gynecologic malignant tumors were separated into 4 cases of epithelial ovarian carcinoma, 2 cases of diffuse ovarian granulosa cell tumor, 2 cases of cervix carcinoma and 1 case of endometrial carcinoma. The mean age was 41.6 years, the intervals from primary tumors to pericardial effusions were 2-49 months. The clinical symptoms were mainly severe shortness of breath. Nine cases were diagnosed by chest X-rays, 6 of them were confirmed by B ultrasonography simultaneously. Three cases were given pericardiocentesis and their cytologic examinations were positive. Two cases of ovarian malignant tumors have been surviving for 49 and 12 months respectively after pericardiocentesis with local chemotherapy and effective systemic chemotherapy, 1 case was survived for 5 months by hormone therapy, but 6 untreated cases were died of cardiac tamponade within one month. CONCLUSIONS: Malignant tumors of genital organs with pericardial effusion could be definitely diagnosed by X-ray, B ultrasonography and cytology. The surviving period may be prolonged by pericardio centesis combined with systemic effective chemotherapy. PMID- 11479963 TI - [Labor induction in women at term with mifepristone and it's safety]. PMID- 11479964 TI - [The development and monitory of cervical intraepithelial neoplasia]. PMID- 11479965 TI - [Diagnostic and therapeutic uses of laparoscopic surgery in the malignant tumor in gynecology]. PMID- 11479966 TI - [Binding of oral lactobacilli to collagen]. AB - The present study surveyed the ability, the characters and the influential factors on lactobacilli binding to the collagen. The experiment employed reference strains and fresh isolated strains. Binding of 3H-labeled bacteria to human type I collagen absorbed on hydroxyapatite surface (C-HA) was tested. The results suggested that the lactobacilli had high affinity for collagen. Competitive inhibitory assay indicated the attachment was highly specific. Binding to C-HA occurred in a collagen dose-dependent manner. pH value of reaction mixtures significantly affected the adherence. Below pH 6.5, increasing pH value caused a reduction of binding. Above pH 6.5, the binding was less affected. The ionic strength apparently influenced the binding level. The results of this study revealed one of the mechanisms that oral lactobacilli target dentin and root surfaces caries. This may also be a cariogenic virulent factor of lactobacilli. PMID- 11479967 TI - [The effect of indirect trauma on the goat temporomandibular joint: a histopathologic study]. AB - The aim of this experiment was to study the effect of indirect trauma on the goat temporomandibular joint (TMJ). Trauma to the TMJ was achieved through an impact to the right mandibular angles of 20 goats, their left TMJs were used as a controls. The animals were killed after 2 h, 1 month, 3 months and 6 months. Light microscopic observations revealed that the condyles exhibited an eburnated, eroded surface with osteophytes, the disks were thinned or perforated, and the temporal surfaces were thickened and the lining were separated. These changes were histologically similar to those of the advanced osteoarthritis. This study suggests that the indirect trauma on TMJ lead to osteoarthritis. PMID- 11479968 TI - [Distribution of leucine-enkephalin like immunoreactive nerve fibers in feline temporomandibular joint: an immunohistochemical PAP method]. AB - In this paper, the immunohistochemical PAP method has been used for the investigation of the distribution of leucine-enkephalin (L-ENK) like immunoreactive fibers in feline temporomandibular joint (TMJ). The results demonstrated as follows: the TMJ capsule, anterior and posterior disc attachments all contained L-ENK immunoreactive nerve fibers which appeared largely as the fine plexi fibers. The density of the distribution of L-ENK positive nerve fibers was the highest in the subsynovium of TMJ capsule, and the plexi structures proceeding to the synovial membrane were observed. The density of the distribution in the lateral joint capsule was the same as the density of distribution in the anterior and posterior discs attachments. No L-ENK positive fibers were detected in the central portion of joint disc. These results indicate that there is the material basis of endogenous analgesic mechanism in TMJ and provide the neuroanatomical basis for exploring physiological functions such as analgesic, immunological and anti-inflammatory effects of ENK nerve fibers in TMJ. PMID- 11479969 TI - [Study on the vaccine against dental caries of recombinant Streptococcus lactis and artificial active immunization. V. Extraction, purification and dot blot analysis of RNA in the recombinant S. lactis]. AB - To investigate the transcription level of the pac gene in the recombinant S. lactis, the amount of PAc-specific mRNA was measured by RNA dot blot analysis with the biotin-labeled 1.5 kb PstI fragment of the pac gene. RNA were extracted with guanidinium thiocyanate followed by centrifugation in cesium chloride solutions. These results indicated the sequence of PAc-mRNA in S. lactis HL107 or HL45 was the same as that in S. mutans Ingbritt and showed similar density photograph. The PAc-specific mRNA plated an intermediary role of protein synthesis in recombinant S. lactis and made it complete the expression of cloned pac gene. PMID- 11479970 TI - [A comparative study of application of PCR technique and Porphyromonas gingivalis nucleic acid probe in clinical examination of apical periodontitis]. AB - By PCR technique and Pg nucleic acid probe, the authors investigated the Pg distribution in root canal samples from patients with clinical apical periodontitis and made a comparative study of the two methods. The results showed that: 1. Pg positive rate of apical periodontitis samples by PCR examination was 74% and that of by nucleic acid probe examination was 76%, and their total coincidence rate reached 94%; 2. between comparative groups there was no significant difference (P > 0.05) in all data by two examination methods; 3. Pg was closely related with clinical symptoms of apical periodontitis such as spontaneous pain, percussion pain, fetidity and periapical abscess. This indicates that the two methods are accurate, quick and convenient and can both be used in direct examination of clinical samples. PMID- 11479971 TI - [The biological effects of recombinant human osteogenic protein-1 to cultured pulp cells]. AB - The study was to observe the biological effects of rhOP-1 on the proliferation, ALPase activity and osteocalcin of dental pulp cells by cell culture technique, MTT, enzyme kinetic methods and radioimmunoassay. The results indicated rhOP-1 could obviously promote the proliferation and enhanced ALPase activity and osteocalcin secretion of pulp cells with showing dose-dependent relationship in this experimental dose range. Thus, rhOP-1 can induce dental pulp cells to differentiate the cells expressing osteoblast phenotype. PMID- 11479972 TI - [Influence of smear layer and adhesive resin on the adaptation of amalgam restoration to cavity walls]. AB - This in vitro study evaluated the effect of removing smear layer and applying adhesive on the marginal adaptation of amalgam fillings to cavity walls. Three groups were established, cavities in group 1 were exposed to 10% EDTA (pH7.4) for 1 minute to remove smear layer before amalgam condensation; cavities in group 2 were also subjected to EDTA, then Miradapt adhesive was applied on the walls, finally amalgam condensed into cavities before the adhesive set; cavities in group 3 received no special treatment before amalgam condensation. After thermocycling, all teeth were subjected to SEM and image analyser. Results showed: Space between restorations and tooth structure in group 1 was 6.31 microns, space in group 2 with adhesive was 3.49 microns and micromechanical interlocking was observed while space in control group was 14.22 microns. Difference among the three groups was statistical significant. It was indicated that removing smear layer and using adhesive resin significantly improved the adaptation of amalgam to cavity walls. PMID- 11479973 TI - [Study of electron microscopy histochemistry and immunohistochemistry of extracellular matrix in adenoid cystic carcinoma]. AB - In order to determine the character of the extracellular matrix in adenoid cystic carcinoma (ACC), 22 cases of ACC of salivary gland were examined by immunohistochemical methods and 4 cases were examined by electron microscopy histochemically. Immunohistochemistry, type IV collagen and fibronectin were more intensively localized in pseudocyst space and in extracellular matrix between tumor cells. Ultrastructural study showed that the pseudocystic cells had abundant rough endoplasmic reticulum and a well-developed Golgi apparatus, some cells had muscle microfilament. There were laminin, proteoglycan and collagen fibril in the pseudocyst space. The results indicated that type IV collagen and fibronectin were the important stromal component in ACC, which may be secreted by neoplasmic myoepithelial cells. PMID- 11479974 TI - [The influence of teeth positions to lower complete denture retention and health of supporting tissues]. AB - Three kinds of three dimension finite element models of edentulous mandibile and the complete denture were established to investigate the influence of artificial teeth positions to denture retention and the supporting tissue. Ten myodynamic and traditional complete dentures were constructed to compare the difference of artificial teeth positions to alveolar ridge. The results indicated: 1. The stress was not obvious different in mandible when teeth were placed over the crest of the ridge or moved lingually or buccally to the ridge in 3 mm; 2. Under occlusion force, it was beneficial for the lower supporting tissue with artificial molars being placed over the slope of mandible and beneficial for denture retention with molars teeth being placed on the crest of ridge buccally; 3. With the effects of oral muscles, the teeth on the crest of ridge buccally or lingually in 3 mm in molar and premolar regions were respectively available; 4. There were obvious difference in arch width of artificial teeth arrangement between traditional and myodynamic complete dentures, the arch was wider in myodynamic complete denture than that of traditional complete denture at molar region. PMID- 11479975 TI - [Determination and analysis of masticatory performance of unilateral free-end removable partial denture]. AB - In this study, the changes of masticatory performance of 30 cases (18 males, 12 females) wearing unilateral free-end removable partial dentures were studied before wearing, 1 week, 1 month and 3 months after wearing by using the light absorption method and peanuts as test food. The results showed: 1. The masticatory performance before wearing unilateral free-end removable partial denture was predominantly lower than that of normal control group. The more teeth lost, the lower unilateral masticatory performance was observed. When the number of lost teeth was few, patients compensated and maintained their masticatory performance before swallowing. 2. The masticatory performance was gradually improved with wearing denture period increasing. 3. Factors relating to the masticatory performance were sex, the extent of alveolar bone resorption and age. Except sex, the extent of alveolar bone resorption and age factors were negative correlative with the masticatory performance; However the number of occlusal units lost, chewing habit and the kinds of denture base materials had no relation to the masticatory performance. PMID- 11479976 TI - [Research on bond strengths of Vita II and plat II ceramic bonding to teeth]. AB - This study compared the tensile bond strengths of Vita II machinable ceramic and Plat II castable ceramic bonding to teeth with three resin cements. The three adhesive cements were Vita cerec, Panavia 21 and TF adhesives. The results showed: 1. The Vita cerec was a good adhesive for Vita II ceramic bonding to enamel and dentine, the bond strengths were 8.91 MPa and 8.48 MPa; 2. The Panavia 21 was suitable for Plat II ceramic bonding to enamel and dentine, the bond strengths were 10.92 MPa and 7.13 MPa respectively. And it was also a good adhesive for Vita II ceramic bonding to enamel, the bond strength was 7.48 MPa; 3. TF had poor adhesive effects for both ceramic materials. PMID- 11479977 TI - [Marginal fit of CAD/CAM ceramic inlays]. AB - The purpose of this in vitro study was to investigate the marginal and internal fit of Cerec-2 inlays before and after adhesion. The test showed: 1. Before luting, the axial marginal space was 67 +/- 18 microns, cervical marginal space was 84 +/- 22 microns and the mean occlusal marginal space was 58 microns. 2. After luting, the thickness of the luting composite was 118 +/- 37 microns at axial margin, and 123 +/- 17 microns at cervical margin, the mean occlusal margin space was 80 microns. This study demonstrated the marginal adaptation of Cerec-2 inlays satisfied the clinical demand, especially at occlusal margin. PMID- 11479978 TI - [Establishment of immediate implant animal model in mongrel dog's fresh extraction socket]. AB - The purpose of the study was to establish the animal models which had clinical characteristics in bone defect sizes around immediate implants. The mesial region of dogs' fresh extraction sockets were made with 3 mm x 3 mm x 5 mm size bone defects around implants with a round bur, then observed their osseous changes at 2 weeks, 4 weeks, 8 weeks, 12 weeks with and without demineralized freeze-dried bone allograft (DFDBA) filled. The results showed that the bone defects in DFDBA grafted group were substituted by new bone, but the defects in ungrafted group were only substituted to 3/5. Such animal models had the advantages of simple preparations and similar clinical characteristics, so they were beneficial to investigation of the immediate implantation. PMID- 11479979 TI - [Amplification, clone and identification of the specific fragments of tumor metastasis-suppressor gene nm23-H1 and nm23-H2]. AB - A series of DNA primers specific for the specific fragments of nm23-H1 and nm23 H2 were designed and synthesized. The specific fragments of nm23-H1(185 bp) and nm23-H2(145 bp) were amplified from human blood DNA by using polymerase chain reaction (PCR). The recovered PCR products were treated with Klenow fragment, inserted into pGEM-3zf(+) vector with blunt-end ligation, and then transformed into competent cell JM109. The positive colonies were directly identified by colour screening on indicator plates. The recombinant plasmids were digested by Alu I and identified by PCR. The results showed that the authors had obtained the specific fragments of nm23-H1 and nm23-H2 respectively, and these specific fragments could be used for study the expression of nm23-H1 and nm23-H2 seperately. PMID- 11479980 TI - [Study of testing velopharyngeal function via the third formant frequency of Chinese vowels]. AB - The value of the third formant frequency (F3) of Chinese vowels of 24 patients with cleft palate and 10 normal children was measured with computerized speech signal processing system (CSSPS), and the rates of velopharyngeal incompetence (RVPI) of Chinese vowels of normal children were quantitatively analyzed using nasopharyngeal fiberscope (NPF). The correlation analysis between the RVPI of vowels [i] [a] and their F3 value of normal group was made. The results showed: 1. The F3 value of Chinese vowels between patients with cleft palate was not significantly different (P > 0.05); 2. The value of F3 in Chinese vowels of normal children was significantly higher than those of patients with cleft palate except [a] (P < 0.05 or P < 0.01); 3. There were generally velopharyngeal incompetence (VPI), in children with cleft palate and velopharyngeal competence of vowels in normal children except [a]; 4. There was highly native correlation between RVPI and F3 in vowel [i], the correlation coefficient was -0.8775 (P < 0.001). It was concluded the F3 of vowel [i] may be one of the most important indices testing velopharyngeal function of postoperative children with cleft palate. PMID- 11479981 TI - [Value of nasopharyngofiberoscope, spectrography and computer in diagnosis and treatment of pathologic speech after cleft palate operation]. AB - The speech of 45 cases with velopharyngeal incompetence (VPI) were evaluated by combining application of nasopharyngofiberoscope (NPF) and computer spectrographic analysis system (CSAS) after cleft palate operation and conservative speech therapy with using cleft palate obturators. The results showed: Improving rate of velopharyngeal incompetence (IRVPI) was close to 33.33% after 2.5-3 years therapy. The spectrographic analysis showed a clearer, and darken frequency band, normal resonance peaks (F2,F3) and seldom appearance of negative value of voice origination time (VOT). The above indicated that patients had achieved adequate velopharyngeal competence (VPC) and good speech habit. It has important significances and greatly improve speech therapy level to apply NPF and CSAS to analysing pathologic speech and evaluating therapic effect. PMID- 11479982 TI - [A study on TF no-mix orthodontic adhesive]. AB - A no-mix orthodontic adhesive composed of primer and paste was developed. The adhesive eliminated the spatulation procedure to which most commercially available products were imperative, could quickly polymerize and cure by means of contact and slight compression of the primer with the paste. It featured high bond strength, convenient handling, enough working time, easy removal of extra adhesive around the bracket and no objectionable odour. The biological tests showed it was safe to use in orthodontic practice, and the results of clinical trials certified the reliable performance, effective bond and convenient use of the adhesive. PMID- 11479983 TI - [Efficacy of a triclosan/copolymer dentifrice in the control of plaque and gingivitis: a six-month study in China]. AB - A total of one hundred fifty three healthy male and female adults were entered into a six month, double-blind clinical study designed to evaluate the effect of a dentifrice containing 0.3% triclosan and 2.0% PVM/MA copolymer in a 0.243% sodium fluoride/silica base, relative to a 0.243% sodium fluoride silica base control dentifrice on supragingival plaque accumulation and gingivitis. Plaque and gingivitis were scored after 3 and 6 months' use of the assigned dentifrice. At the end of the study, the dentifrice containing triclosan/copolymer demonstrated an average reduction, relative to the control dentifrice of 16.1% in plaque accumulation and 24.3% in gingivitis. Both were statistically significant(P < 0.001). It was concluded that the twice-daily use of the triclosan/copolymer containing dentifrice resulted in a significant reduction in plaque formation and a significant improvement in gingival health. No side effects were observed. PMID- 11479984 TI - [Effects of specific antibodies against the cell surface protein P1 of streptococcus mutans MT 6R upon adhered streptococci]. AB - The sera and saliva were obtained from BALB/C mice by injection of the cell surface protein P1 and Freud's adjuvant. High antibody titers were showed in the sera and saliva by an enzyme-linked immunosorbent assay. The cells of S. mutans (serotype c, e and f) adhered on saliva-coated hydroxyapatite could be detached by the antibodies of the protein P1 of S. mutans MT 6R including in the sera and saliva (P < 0.05), but the antibodies had no influence on the cells of S. cricetus AHT, S. sobrinus OMZ176, S. sobrinus 6715 and S. rattus BHT (P > 0.05). PMID- 11479985 TI - [Effects of different dilution of specific antibodies against the cell surface protein P1 of S. mutans MT 6R upon its adherence]. AB - The sera and saliva were obtained from BALB/C mice by injection of the cell surface protein P1 and Freud's adjuvant. High antibody titers were showed in the sera and saliva by an enzyme-linked immunosorbent assay. Different dilution of the specific antibodies against the protein P1 of S. mutans had different inhibition effects on adhering of S. mutans MT 6R. Adherence of streptococcus mutans MT 6R on saliva-coated hydroxyapatite was reduced significantly by using sera and salivary antibodies in dilution of 1:1, 1:2, 1:4, 1:8, 1:16, 1:32 (P < 0.05). PMID- 11479986 TI - [Histologic and SEM studies on aging changes of temporomandibular articular disc in young growing rats]. AB - The normal aging changes of the temporomandibular articular discs in young growing SD rats were studied with histologic and scanning electron microscope methods. The results showed that the active remodeling growth was found in the discs of the rats from 3 to 9 weeks, which characterized as the changes from the young cellular connective tissues to fiberous cartilage gradually, these changes were consistent and adaptable with the growth of the condyle. The wave-like structures on the superior surface of discs of the rats from 5 to 8 weeks were varied morphologically in different bands and ages. PMID- 11479987 TI - [Histologic and histochemical studies on the temporomandibular articular disc in young growing rats after functional mandibular advancements]. AB - The purpose of these studies was to detect the patterns of the changes in the site of temporomandibular articular disc in the young growing SD rats after the functional protrusive appliance inserted with histologic and histochemical methods. The results showed that collagen fibres were dense, roughly paralleled linked especially anterioposteriorly after the mandible protruded functionally, chondrocytes and gycosaminoglycans contents increased apparently in the posterior band of the discs. The adaptive remodeling of the discs might be the results of the pressure of the condyle on the discs and the lateral pterygoid muscle to it. The remodeling growth of the discs were coordinate and adaptive with the proliferative changes of the condylar cartilage after the mandible protruded functionally. No pathological changes were found in the rat articular discs. PMID- 11479988 TI - [Immunohistochemical study on the temporomandibular articular disc in young growing rats after functional mandibular advancements]. AB - In the present experiment the type I collagen of discs was investigated with immunohistochemistry in the young growing SD rats after the mandible protruded functionally. The experimental evidence showed that the immunohistochemical staining reactions accentuated with age changed in control group discs, and the immunohistochemical staining reactions in experimental group were intenser than in control group. It indicated that the discs synthesized higher contents of type I collagen after the disc accepted more functional stimuli in the experimental group. PMID- 11479989 TI - [The mechanisms of antioxidantenzyme against inflammatory lesion in animal oral mucosa. I. An investigation on microcirculation in oral mucosa]. AB - A technique of vital microcirculation observation and electron microscope were used to study the microcirculation in hamster cheek pouch, in which an acute inflammatory lesion was induced by oxygen free radicals (OFR). At the same time, the effect of superoxide dismutase (SOD) on the microcirculation was assessed. The results showed that in the model group the microcirculation disturbance in the cheek pouch was very serious. However, in the SOD group the microcirculation was almost unchangable. PMID- 11479990 TI - [A comparative study of soft tissue cephalometric of normal occlusion between the middle-old-aged people and the young people]. AB - In this study the soft tissue cephalometric data of normal occlusion between 39 middle-old-aged people and 30 young people were compared. The results indicated: the upper lips of adults were getting thinner and longer, the soft tissues of lower lips and chins were getting thicker, so that the profiles changed obviously with the increasing of ages. PMID- 11479991 TI - [The culture of TMJ condylar cartilage cells and study on their biological behaviors in vitro]. AB - To investigate the behavoir of TMJ condylar cartilage cells in vitro, the mandibular condylar cartilage cells were harvested from a 5-month-old human fetus by dissection and sequential digestion with 0.25% trypsin and 0.2% collagenase (type II). The isolated cells were cultured in DMEM medium and identified by histochemical and immunohistochemical methods. Cell proliferation, morphology and ultrastructure were observed by phase-contrast microscope, cytologic staining and electronic microscope. In primarily cultured cells, polygonal chondroblast-like cells dominated and they were confirmed by the positive result of immunohistochemical examination for type II collagen and Toludin blue staining. In conclusion, the TMJ cells in this culture system kept their phenotype in vivo. PMID- 11479992 TI - [Comparison of the activity for carbohydrate metabolism enzymes of plaque in different ages]. AB - Plaque samples were collected from tooth surface by the sampling method in 28 volunteers (two groups, 14 young people, ages ranged from 19-26 years, mean: 22.29 years old; 14 old people, aged between 60-70 years, mean: 64.35 years old, DMFT > 3). The activity for three enzymes including amylases, LDH, GTF was tested by micro-chemistry enzyme analysis method. The results showed that there were no significant difference for the activity of three enzymes in two groups (P > 0.05). The finding suggests that dental plaque is a specific microecologic environment and cariogenic factor, age does not affect the activity of enzymes, plaque in different ages has similar cariogenicity. PMID- 11479993 TI - [Analysis of apoptosis in premalignant and malignant epithelia of buccal mucosa]. AB - To determine the relationship between apoptosis and carcinogenesis of oral mucosa, the distribution patterns of apoptosis in normal (N), inflammation (IF), dysplasia (DYS) and squamous cell carcinoma (SCC) of oral mucosa were evaluated by in situ end-labeling method. Results showed that the apoptosis levels in N and IF were relatively lower and stable, whereas the apoptosis of DYS was the highest one in all observed groups. The apoptosis of DYS and SCC were increased obviously higher than N and IF. The results indicated that apoptosis played an important role in premalignant lesion and malignancy of oral mucosa. PMID- 11479994 TI - [A three-dimensional anisotropic finite element analysis of modified bar-clip implant-borne overdenture in the edentulous mandible under punching loads]. AB - The stress distributions of the modified bar-clip implant-borne overdenture in the edentulous mandible were performed under punching loads with both porcelain and resin chosen as the restorative materials by means of three-dimensional anisotropic finite element method. The purpose of this study was to investigate the patterns of stress distributions and to compare the effect of different restorative materials on the stress distributions under punching loads. The results showed that the value of stress peaks in implants with porcelain restorations seemed higher than those with resin restorations. On the contrary, in the bone-implant interface had little difference, except a comparative uniform stress distribution found in resin restorations. It implies that resin restoration produces a cushioning effect on reducing the stress peaks in implants themselves under punching loads. Both porcelain and resin restorations can be chosen as the restorative materials because of a little influence on the stress peaks in bone-implant interface under punching loads. PMID- 11479995 TI - [Early observation of healing of rabbit mandibular defects with porous blocks consisting of polylactic acid and recombinant human bone morphogenetic protein 2]. AB - The effects of bone morphogenetic protein (BMP) which can induce bone formation have been surely proved, Porous polylactic-acid blocks were prepared by a freeze drying method. During preparation of the blocks, recombinant human bone morphogenetic protein-2(rhBMP-2) was mixed with the polylactic-acid(4 mg per block). PLA-rhBMP-2 was implanted in the mandibular defects of rabbit and retrieved after 2 or 4 weeks. The results showed that: with the PLA-rhBMP-2 blocks, new bone formation was observed in 2 weeks after implantation, and plain PLA blocks produced a little of new bone formation in the edges of defect even in 4 weeks after implantation. Calcium content of the retrieved PLA-rhBMP-2 block was statistically higher than that of plain PLA blocks. These results suggest that PLA may be effective delivery system for BMP and the PLA/rhBMP-2 block may potentially have usefulness as a bone graft substitute. PMID- 11479996 TI - [A biomechanical study on the bone repairing ability of recombinant human bone morphogenetic protein-2-coral composited artificial bone]. AB - To improve bone repairing ability of coral, recombinant human bone morphogenetic protein-2 (rhBMP-2)-coral composited artificial bone was obtained by combining coral with rhBMP-2 in this experiment. The composite was implanted into rabbit calvarial critical-size defects and coral alone was implanted as control. The bone repairing ability was assessed by radiography, histology and biomechanics. The results showed that implants were absorbed gradually after they was implanted into the defects. In the meantime, the new bone was formed and the mechanical strength was reinforced uninterruptedly in bone defect regions. At 12 weeks, the implants were replaced completely by bone, and the mechanical strength was entirely restored. These suggest that the composite possess a superior bone repairing ability and may be one of the most perfect bone graft substitutes currently available. PMID- 11479997 TI - [Determination on the electric-mate order of prosthodontic materials in artificial saliva]. AB - Electric-mate corrosion of the different prosthodontic materials has been noticed in oral cavity. The aim of this investigation was to determine corrosion potential of prosthodontic materials, and to acquire the electric-mate order in artificial saliva. That will conduct us to select the materials of resistance corrosion. For resistance electric-mate corrosion, it's better to use same materials, or to use close materials of the electric-mate order. PMID- 11479998 TI - [The preliminary study on WCMM machinable glass-ceramics]. AB - The aim of this work was to investigate machinable glass-ceramics used as oral restoration material in K2O-MgO-Al2O3-SiO2-F glass system. By using of DTA, XRD and SEM investigation, the crystalline structure and characteristics were tested. The chemical composition of WCMM machinable glass-ceramics was defined by the following general formula: (K,Na)Mg3-3x Al2x (AlSi3) O10F2(0.5 < or = x < 0.7). The experimental results show that WCMM is an excellent oral restoration materials in the fabrication of crowns, inlayers and veneers. PMID- 11479999 TI - [Experimental study on fitness of retainers of one-piece casting frameworks of distal-extension implant denture]. AB - An experiment on fitness of retainers of one-piece casting frameworks was performed between two non-noble metals and among three of abutment positions fixture implant with a coordinate measuring machine for three-dimensional analysis. The results showed that there was no statistically significant differences in retainer fitness between the two metals, and among the three fixture abutment positions. However, all retainer displayed shrinkage in three dimensions, and the retainer shrinkage on the cervical surface were more than on the occlusal surface. The fitness of retainers of one-piece casting frameworks would be improved if used in multi-unit implant fixed denture. PMID- 11480000 TI - [Detection of point mutation of p53 gene in head and neck squamous cell carcinoma by non-isotopic PCR-SSCP]. AB - Non-isotopic PCR-SSCP technique was used to detect point mutation of p53 gene in fresh tissue taken from head and neck squamous cell carcinoma. The abnormal band of the single-stranded DNA (ssDNA) was identified in 7 of 13 cases. The results showed that point mutation occurred in the related DNA fragments. The point mutation of exon 5 was found in one case and that of exon 6, 7, 8 in two respectively. These results indicated that the p53 gene mutation was closely correlated with the development of head and neck squamous cell carcinoma. Compared with the traditional isotopic PCR-SSCP technique, the non-isotopic PCR SSCP one is a kind of simpler, safer, quicker and more effective way to detect the mutation of p53 tumor suppressor gene. It is especially useful in the screening of point mutation when handling a large number of samples. PMID- 11480001 TI - [Role of type IV collagene and type IV collagenase in the invasion and metastasis of salivary adenoid cystic carcinoma]. AB - Using immunohistochemical ABC method, the authors studied the distribution of type IV collagen and type IV collagenase in 25 cases ACC. The result showed type IV collagen was found in vascular and epithelial basement membrance, and in partly cyst of sieve-like and tubular pattern, and in cytoplasm of tumor cell. Type IV collagenase was detected in membrance and cytoplasm of tumor cell. Expression of type IV collagen and type IV collagenase were correlated with pathologic type, TNM stage and metastasis. Much positive staining of type IV collagen was found in sieve-tubular pattern, early stage of TNM and without metastasis, but much positive staining of type IV collagenase was found in solid pattern, later stage of TNM and metastasis. The result suggested that the loss of type IV collagen and over-expression of type IV collagenase may be markers of malignance of ACC. PMID- 11480002 TI - [Effect of complete dentures on the blood flow and blood cell velocity of alveolar mucosal microcirculation]. AB - The Purpose of this study was to determine the effects of complete dentures on alveolar mucosa microcirculation. The alveolar mucosal microcirculation of 17 edentulous subjects was measured by videomicroscope. The variance of the alveolar mucosa microcirculation before and after insertion of the dentures was compared. The results showed that: 7 days after wearing complete dentures, the mucosal blood flow and blood cell velocity of maxillary anterior alveolar ridge crest decreased significantly (P < 0.05), those of labial region of maxillary anterior alveolar ridge increased significantly (P < 0.05), 14 days after wearing complete dentures, the mucosal blood flow and blood cell velocity almost recovered the levels before wearing the dentures. The results can be relative to adjustment the occlusion in the early period of wearing the complete dentures and not wearing the complete dentures at night. PMID- 11480003 TI - [Study on diagnosis of oral mucosal melanoma with histology, electron microscopy (EM) and immunohistochemistry]. AB - Fifteen cases of primary oral mucosal melanoma were studied with the methods of Fontana Staining, EM, and immunohistochemistry. The results showed that 4 of 15 cases were non-pigmented lesions. In these cases, two were positive with Fontana staining. Pigment body could not be found in three cases observed under EM, but all the 15 cases were positive with immunohistochemical staining. The results suggested that the immunohistochemical staining should be done for those cases suspected as non-pigment melanoma. PMID- 11480004 TI - [A comparative study of skeletal cephalometric of normal occlusion between the middle-old-aged and young people]. AB - In this study the skeletal cephalometric data of normal occlusion between 39 middle-old-aged people and 30 young people were compared. The results indicated: the growth and reconstruction of the skeletal tissues of skull and jaws and face could still maintain for a period of time after grown-up. It mainly showed that skeletal tissues increased in vertical direction, the declination of occlusion plane became flat along with the increasing of ages. PMID- 11480005 TI - [The clinical analysis of eosinophil infiltration in human squamous cell carcinoma of buccal mucosa]. AB - Tumor-associated tissue eosinophils (TATE) is a host's defensive reaction to tumor. In this study, authors examined the eosinophil of 40 squamous cell carcinomas of buccal mucosa. Infiltration of the primary tumor by eosinophil was found to have marked prognostic significance. High-grade TATE was associated with the absence of head and neck lymphonode's metastasis (P < 0.05). There was a statistical significance (P < 0.05) that high-grade TATE was a favorable prognostic indicator for squamous cell carcinoma of the buccal mucosa. Authors concluded that quantitative assessment of eosinophils was one of the most important aspects of the microscopic evaluation of this human carcinoma. PMID- 11480006 TI - [Preliminary evaluation of plastic crown restoration supported by osseointegrated implants]. AB - This study was to evaluate the feasibility of plastic crown restoration supported by osseointegrated implants. The following conclusions were drawn from this study: plastic crown gave better biomechanical consideration than porcelain fused to metal (PFM) crown in osseointegrated prostheses, but plastic crown gave worse wearability, tensile strength, compression strength and flexuaral strength than PFM crown. After restoration the disadvantages of the plastic crown were beyond the clinical acceptable range. It showed plastic crown designed dental prothetic implantation was unfeasible. PMID- 11480007 TI - [Fabrication and use of two kinds of overdenture attachments]. AB - The authors introduced the fabrication of stud attachment and bar attachment used to increase the stability and retention of overdenture. During two years' periods, the results of the clinical practice indicated: the retention of overdenture was increased and patients' masticator function was improved when attachments were used, and then the fabrications of two kinds of attachments were simple and value of attachments is cheap. The technique was popularized and used easily. PMID- 11480008 TI - [An evaluation of the long-term results of palatopharyngeal closure function after velopharyngeal ring ligation]. AB - 16 cases were selected randomly from a group of 36 postoperative patients who underwent velopharyngeal ring ligation 16 years ago for evaluation of pronunciation, pharyngeal radiography, and electric endostroboskop to evaluate the patients' velopharygeal closure function. It was found that the pronunciation function of 93% patients were in good status. The results showed that centripetal force of velopharyngeal ring ligation made the velopharyngeal cavity reduced from the right to left and from anterior to posterior sides. It was good for sphincter of velopharyngeal moving, which could reduce the space of pharyngeal cavity and add the length of soft palate. The velopharyngeal closure was improved, which produced a good condition for the recovery of pronunciation function, at the same time, the change of pharyngeal cavity adapts to the patient's growth without limiting the growth and development of nasovelopharyngeal. PMID- 11480009 TI - [Oral candidiasis in Sjogren's syndrome: prevalence, clinical features and treatment]. AB - The clinical feature of 20 cases with oral candidiasis in Sjogren's syndrome (SS) were reported, and the treatment effect by topical nystatin smearing, 2% sodium bicarbonate solution rinsing, and transfer factor injecting for about 2 months were analysed. The results showed that the diagnostic criterion of oral candidiasis in SS included two aspects: the SS changes confirmed by labial gland biopsy and candidal hyphae and spores found in a smear. The results indicated that the comprehensive treatments were proved to be effective for oral candidiasis in SS. PMID- 11480010 TI - [Benign focal melanotic lesions of the oral mucosa: a clinicopathologic study of 41 cases]. AB - 41 patients with benign focal melanotic lesions on the oral mucosa were studied. The results showed that among the patients, 27 were female and 14 male, the mean age was 33 years old, and the most common site was the lip and gingiva. Clinically, the lesions were focal and well-defined, the size was about 0.6 cm in diameter. Histologically, 21 cases were pigmented nuvus on the oral mucosa (18 cases with nevocellular nevus and 3 cases with the common blue nevus), 2 cases were junctional change, 5 cases were the lentigo simplex, and 13 cases were oral melanotic macule. The characteristics of the oral melanotic macule and the junctional change were discussed. PMID- 11480011 TI - [A study on the factors affecting the result of correction of saddle nose with implant of artificial bone]. AB - This study made statistics of 816 cases of correction of saddle nose with implant dense polycrystalline hydroxylapatite partic artificial bone. The factors which affected the result of the operation included selection of graininess of the artificial bone, design of implanting bed and operating errors, etc. These factors were studied in detail in this paper. PMID- 11480012 TI - [A study on the treatment and restoration of the complete fractured teeth]. AB - In order to keep the fractured teeth with their important function of mastication the authors had adopted a kind of improved calvital preparation to carry out the pulp treatment for 56 cases, 63 teeth, together with periodontal washing, the application of medicine and the periodontal treatment of gingivectomy and flap operation, etc. from Jan. 1989 to Jan. 1993. The regularization of the filling of the fragmental area, the combination and the fixation of the split tooth were strengthened with spiral root post and dentin retaining nails, and then the full cast crown was restorated. The results were satisfied. It turns out that complete fracture of posterior teeth, thought to be pulled out in the past because they had split well below the gum, or even affected the pulp chamber floor, can be kept if given treatment and proper restoration. PMID- 11480013 TI - [The effect of cryotreat on the size of the medium and high melting-point castable alloys]. AB - Castable alloys are widely used in prosthodontics. The properties of alloys can be improved by cryotreat. This method may cause a size alteration of materials. This study revealed the effects of different cooling rates on the size of the castable alloys. The results showed: 1. The rapid cooling rate caused significant deformation of the alloys. 2. The 3 degrees C/min rate and the 1 degree C/min rate did not cause any significant deformation of the alloys. 3. The 3 degrees C/min rate and the 1 degree C/min rate did not show any difference in their deformation causing ability. 4. The rapid cooling rate and the slow cooling rate (3 degrees C/min and 1 degree C/min) showed significant difference in their deformation causing ability. These results suggested that 3 degrees C/min was recommendable as a cooling rate of the cryotreat of the medium and high melting point castable alloys. PMID- 11480014 TI - [The effect of cryotreat on the mechanical properties of the medium and high melting-point castable alloys]. AB - Two mechanical properties that are closely related to the clinical practice, i.e. the surrendering toughness and the hardness, of the medium and high melting-point castable alloys, were tested before and after the cryotreat. The results showed: 1. Cryotreat significantly increased the surrendering toughness of the medium and high melting-point castable alloys (40.00%); 2. Cryotreat significantly increased the surrendering toughness of the medium and high melting-point castable alloys (25.45%); 3. Cryotreat significantly increased the surrendering modulus of the high melting-point castable alloys (25.77%); 4. Cryotreat significantly increased the hardness of the medium melting-point alloys(24.40%); 5. Cryotreat significantly increased the hardness of the medium melting-point alloys (7.77%). These results suggest that cryotreat is an effective procedure in enhancing the toughness and hardness of the medium and high melting-point castable alloys. PMID- 11480015 TI - [Antitumor effect of TIL combined with radiotherapy on mouse uterocervical cancer U14]. AB - After yielding culturing mouse TIL from U14 with Rosenberg's technique, the authors observed the antitumor effect of TIL combined with radiotherapy on established mouse transplanted tumor of uterocervical cancer U14. The results demonstrated TIL treatment or radiotherapy could inhibit the tumors growth. Combination showed a stronger inhibition and the E/O value was 1.1. It implied that an additive effect was gotten when combining TIL with radiotherapy. PMID- 11480016 TI - [Dynamic distribution in vivo of TIL from human tongue cancer after local injection in animal model]. AB - On the basis of established nude mice transplanted tumor model of human tongue squamous cell carcinoma, peritumor injection was performed with 3H-TdR labeled TIL from human tongue cancer after 4 weeks culture. The dynamic distribution of 3H-TIL in vivo was learned through detecting the radioactivity of the target tissues. The results demonstrated that 3H-TIL was found in the whole body. There were about 2.1-30 times more in tumor tissues than in normal tissues and 3H-TILs even collected in the tumor tissues distal to the injective sites. The nearer to the injective sites the tumor tissues, the more 3H-TIL collected. It showed that local injection of TIL could be expected to exert a better antitumor effect. PMID- 11480017 TI - [The development and application of computer assisted photogrammetric-system of facial soft tissue running under multiple facilities for taking image]. AB - The photogrammetric-system was developed by using multimedia computer, colour television camera, SE100 video blaster, Newsketch 1212 HR digitizer and by using Visual Basic language. It ran under Windows 3.1 and Windows 95. The features of this system: 1. It had three kinds of functions: photogrammetric function for frontal and profile facial soft tissue, function for managing file and function for forming dynamic data base. 2. It ran under different facilities for taking image (television camera with video blaster, digitizer, scanner) speedily and specificly, and had very friendly interface, and was of multimedia. The application of this system: 1. assisting diagnosing, making plan for treatment and observing results of treatment in clinic of orthodontics, orthopedics and facial plastics, 2. assisting analysing the features of facial soft tissue about certain people, 3. assisting the multimedia teaching. PMID- 11480018 TI - [The analysis of frontal facial soft tissue of normal native adult of han race of Guangdong province by using the computer assisted photogrammetric-system]. AB - The authors analysed the feature of frontal facial soft tissue of 150 native adults of Guangdong province by using the computer assisted photogrammetric system made by the authors, which contained several new mode parameters planed by the authors. THE RESULTS: 1. The mean measurements of six parameters (Sn-Mes, Gs Mes, Exr-Exl, Enr-Enl. [symbol: see text]Zyr-Rc-Mes, [symbol: see text]Zyl-Lc Mes.) of the male subjects were larger than those of the female subjects, and the mean measurements of two parameters (Zyr-Zyl/Gs-Mes, Rc-Lc/Gs-Mes) of the male subjects were smaller than the ones of the female subjects. 2. The four measurements (Tr-Mes, Gs-Sn, Zyr-Zyl, Chr-Chl) in both male and female subjects of Guangdong province were smaller than the ones of Sichuan and Shandong provinces, and one measurement (Tr-Gs) of Guangdong male subjects was also smaller than the one of Sichuan or Shandong province, and one measurement of Guangdong female subjects smaller than the one of Shandong province. The conclusions: 1. The normal native adults of han race of Guangdong province show sex-difference and region-difference in the feature of frontal facial soft tissue. 2. The new-mode parameters (such as: [symbol: see text]Zyr-RC-Mes, [symbol: see text]Zyl-Lc-Mes, Zyr-Zyl/Gs-Mes, Rc-Lc/Gs-Mes) are helpful in studying the feature of frontal facial soft tissue. PMID- 11480019 TI - [An inquiry on the normal value of Chinese' ENoG and the influencing factors]. AB - The normal value of the ENoG of 172 facial nerves from 86 Chinese has been estimated with the method of regulating the strength of stimulating current. It showed that the CAPs of the Chinese' ENoG were of normal distribution. The mean of ampitudes was 3.94 +/- 1.02 mV. The coefficient of variation was 25.8%; The mean of latencies was 3.18 +/- 0.42 ms. The coefficient of variation was 13.2%. The variance from age to age, sex to sex and side to side were not significant. It was concluded that the interpretation of ENoG should be based on the fact that the CAP obtained from one certain subject was symmetrical between the opposite sides. The effecting factors which influenced the ENoG of normal facial nerve were also discussed in this paper. PMID- 11480020 TI - [An experimental study of injury characteristics in maxillofacial region following different kinds of high velocity projectiles wound]. AB - Model 7.62 mm, model 5.56 mm bullets and 1.03 g steel sphere, 1.0 g fragment with impacting velocities 1300-1400 m/s were used to shot soap target, mandibular area of pigs in vitro and dogs in vivo. The shape and size of cavitations and wounds were observed and the specimens of wounded muscles were collected for light and electron microscopic observation. The authors found that different kinds of cavitations formed in different kinds of projectiles wound, which produced different injured characteristics in maxillofacial firearms wound. The characteristics of maxillofacial wound ballistics and difference of maxillofacial wounds in different kinds of high velocity projectiles wound were also discussed in the article. PMID- 11480021 TI - [Preliminary study on the effects of Nd-Fe-B magnet on trace elements in rhesus monkeys]. AB - Three adult rhesus monkeys were selected. Two of them were fixed with Nd-Fe-B magnets on the palates respectively. Nd-Fe-B magnet was not used in one monkey. Blood was drawn from all monkeys. Ten kinds of trace elements in blood were measured. The result showed that there was no quantitative difference in ten kinds of trace elements between the experimental and control animals. PMID- 11480022 TI - [Effect of stud attachment post design on dentin stress distribution]. AB - The shape design of attachment post is crucial factors influencing post retention and stress distribution to the dentin. By means of CAD and FEA, the present study investigated the effects of the shape designs of attachment posts on the stress distribution in dentin. The results indicated that the taper post had wedging effects on the dentin and produced higher Von Misses stress than that of parallel side post and the Von Misses stress value of the dentin was about 1.1 times. The maximum Von Misses stress value that the design of the thread post produced was about 1.06 times the stress of the nonthread post did. However, the effects of post design on the dentin were minor compared with the bite force directions. In the same post design, the maximum Von Misses stress in dentin with 45 degree oblique force was about 2.1 times with vertical force. Therefore, reducing oblique force is the main method to prevent root fracture in clinic. PMID- 11480023 TI - [Preliminary determination of the properties of HX-I refractory die material]. AB - The setting time, the setting expansion ratio and the compatibility with impression materials of the HX-I refractory die material were determined. The results showed: the setting time was 11 +/- 3 min: the setting expansion ratio was 0.23%; the compatibility with impression materials was excellent between HX-I die material and agar or silicone rubber respectively, but the compatibility was bad between HX-I die material and alginate. These properties of HX-I die material were close to the ones of Lamina die material. PMID- 11480024 TI - [P53 expression and clinical significance in oral squamous cell carcinoma]. AB - P53 protein expression was investigated in oral squamous cell carcinomas and oral premalignant lesions by monoclonal antibody Do-1 and immunohistochemistry technique. 4 of 12 (33.3%) samples of severe epithelial dysplasia and 25 of 44 (56.8%) samples of squamous cell carcinoma expressed P53 protein while all the normal mucosa, mild and moderate epithelial dysplasia were negative. The P53 expression in carcinomas was associated with differentiation, lymph node metastasis and tumour stage. This result indicated that P53 genic mutation might be an early event in oral mucosa carcinogenesis and related to oral tumor progression. Detection of P53 protein probably has clinical significance in identifying the premalignant lesions of oral mucosa and predicting the prognosis of oral carcinomas. PMID- 11480025 TI - [The study on analgesic effect of local-release delivery system--Huangjiabang]. AB - Laboratory investigation was made on analgesic effect of local release delivery system, Huangjiabang, which mainly includes Chinese medical herbs. Through the application of twisting-body and thermal-plate experiments on white rats. The result showed that the pain threshold was markedly elevated (P < 0.05, P < 0.001) after the rats were fed with Huangjiabang 125 mg/kg or 250 mg/kg body weight, and the number of twisting body's animals significantly decreased (P < 0.001). The analgestic mechanism of Huangjiabang delivery system relates to its effect of invigorating the circulation of blood and accelerating the stasis assimilation, remitting the inflammation. PMID- 11480026 TI - [A study on the significance of p53 gene mutation, P53 protein positive staining and PCNA staining]. AB - After obtaining the results of p53 gene mutation by a silver staining method to polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, P53 protein staining and proliferation cell nuclear antigen (PCNA) index by double-blind method, multivariate correlation analysis two by two showed that PCR-SSCP, P53 positive protein staining and PCNA index had markedly correlation not only in oral precancerous lesions (OPL) but also in primary sites and regional metastatic lymph nodes of oral squamous cell carcinomas (OSCC). This indicated that the nature of the three indexes was the same. After introducing the sex and age of patients which have been proved to be related to the initiation and development of OPL and OSCC according to other researchers and are easy to be obtained in clinics, multivariate discriminatory analysis was carried on and three groups of discriminatory equations were gotten about the pathological grading of OPL and OSCC and metastasis condition of OSCC. The accuracy of these equations was 82.5%, 78% and 80%, individually. This suggests that the equations have applicable potentiality already. PMID- 11480027 TI - [Effect of human neutrophil defensins on subgingival flora]. AB - 40 teeth of 18 patients with moderate or severe adult periodontitis were examined for evaluating the effect of defensins on subgingival flora. The period of experiment was 11 days. Defensins fibers were put in the periodontal pockets deeper than 5 mm for 3 days and discarded on the fourth day. Each of the pockets was applied one fiber containing about 8 micrograms defensins. The clinical parameters (GI,BI) and the density of subgingival flora were reduced (P < 0.05). The increase of coccoid cells, and the decrease of spirochetes and rods were indicated in the relative ratio (P < 0.05). This study indicated that defensins could reduce the density of the subgingival flora and change their ratio towards the periodontal healthy flora level. PMID- 11480028 TI - [Soft tissue profile changes following surgical correction for Chinese adults with maxillary protrusion]. AB - The soft tissue profile response to anterior maxillary osteotomy for anterior maxillary excess patients was evaluated. Soft tissue change to hard tissue change ratios as well as linear regression equation were calculated from a patient pool representing fifteen anterior maxillary osteotomies. The soft tissue profile response to anterior maxillary osteotomy appeared to be predictable. The results led to the following abbreviated conclusion: posterior movement of subspinale was accompanied by posterior movement of 37% (r = 0.73) at subnasale, 63% (r = 0.87) at superior labial sulcus. Posterior movement of incision superius was accompanied by posterior movement of 75% (r = 0.86) at labrale superius. PMID- 11480029 TI - [A study of craniofacial type of skeletal anterior crossbite in juveniles]. AB - Lateral skull radiographs of 69 subjects with Class III malocclusion, 35 boys, 34 girls, aged 10-13 years old, were compared to a control group of similar mean age based on hierarchical cluster analysis. Cranial base, maxilla, mandible, relationships of sagital and vertical, teeth and soft tissue were evaluated together. Four subgroups of malocclusion were identified and described at the first time. The subgroup of maxillary deficiency, mandibular prognathism due to moved forward joint and lower anterior face height was the most common type. The results indicated that it should be evaluated a lot of indices together on cephalometric analysis. PMID- 11480030 TI - [The modified RPI clasp design]. AB - The modified RPI clasp was described and compared with the conventional RPI clasp in the article. The modified RPI clasp could provide adequate retention with the minimum undercut of the most distal part of abutment tooth and gain good esthetical effect. The I bar tip would disengage from the tooth and reduce or avoid torsional forces transmitted to the abutment tooth when rotation of distal extension base occurred around the mesio-occlusal rest. Therefore, it is a new type of clasp design for the distal-extension removable partial dentures. PMID- 11480031 TI - [Facial proportions of the young with the beautiful faces of Sichuan descent]. AB - In this study, eighteen indexes of facial proportions were established in 72 youngsters with beautiful faces of Sichuan descent, who haven't been done operation on and orthodontics in our country. The results indicated that there were not most sexual difference of the facial proportions between male and female in most indexes, and also were not national difference of facial proportions between Chinese and Japanese. It also showed some progresses in clinical practice. PMID- 11480032 TI - [Effect of wire ligature splint reinforced with preparing groove and employing composite materials on the teeth with severe periodontitis]. AB - Teeth with severe periodontitis exhibit alveolar bone loss, teeth mobility and even teeth loss. The author made groove along the buccal and labial surfaces of teeth, twisted the stainless steel wire and placed ultraviolet light polymerized composite resin over the wire. Teeth with mobility 2-3 degree from 39 patients were treated and the effect was evaluated in the period of 2 to 4 years. The results demonstrated that the treatment had advantages of stabiliting very loose teeth and forming a firm unit. It minimized teeth mobility and improved greatly teeth occlusal function. When occlusing, the device could redirect force to other teeth, thereby reduced traumatism and facilitated periodontal tissue repairing and regeneration and protected loose teeth. PMID- 11480033 TI - [A probe into rehabilitating occlusion of majority teeth missing]. AB - It is generally to be found that decrease of occlusal vertical dimension, worn occlusive surface of remaining teeth without opposite teeth and loss of intercusping position (ICP), etc. in the patients with majority teeth missing. In western countries, fixed restorations (including implant dentures) are routinely used for the patients with majority teeth missing in order to rehabilitate occlusion. The authors applied occlusal theory, combining with the clinical facilities and used one-piece casting base and functional impression technique for rehabilitation of occlusion. The purpose of this article was to summarize clinical experiences, to explore the relative problems concerning rehabilitation for patients with majority teeth missing. PMID- 11480034 TI - [mRNA expression of metastasis-suppressor gene nm23 in carcinoma of buccal mucosa I. Northern blotting study]. AB - In this study the authors detected both nm23-H1 and nm23-H2 mRNA levels in 52 tissues samples of patients with carcinoma of buccal mucosa (CBM) by Northern blotting method. The expression levels of both nm23-H1 and nm23-H2 mRNA in CBM were higher than those in normal buccal mucosa, leukoplakia, adjacent nontumorous mucosa, and lymph nodes with or without metastasis. The nm23-H1 mRNA expression levels in CBM with lymph nodes metastases were lower than those in CBM without metastasis, and were lower in metastatic lympho nodes (P < 0.05). Of the 11 cases with metastases, 9 cases (81.8%) showed low expression of nm23-H1 mRNA, of 19 cases without metastasis, 15 cases (78.9%) were high expression. No significance of nm23-H2 mRNA expression levels were found between CBM with and CBM without metastasis (P > 0.05). The expression of nm23-H1 and nm23-H2 mRNA showed no significant correlation with each clinical-pathological parameter examed. The result suggested that the expression of nm23-H1 mRNA significantly correlated inversely with lymph node metastasis in CBM, while the expression of nm23-H2 mRNA not. nm23 mRNA expression levels can provide a valuable predictor of lymph node metastases in CBM. PMID- 11480035 TI - [mRNA expression of metastasis-suppressor gene nm23 in carcinoma of buccal mucosa. II. Quantitative reverse transcription PCR amplification]. AB - The nm23 gene is a conspicuous metastasis-suppressor gene. The authors detected both nm23-H1 and nm23-H2 mRNA levels in 47 tissues samples of patients with carcinoma of buccal mucosa (CBM) by quantitative reverse transcription PCR amplification. The results showed that expression levels of both nm23-H1 and nm23 H2 mRNA varied in normal buccal mucosa, leukoplakia, adjacent nontumorous mucosa, submandibular gland, CBM and lymph nodes with or without metastasis. The nm23-H1 mRNA expression levels in CBM with lymph nodes metastases were lower than those in CBM without metastases (P < 0.05), while no significance of nm23-H2 mRNA expression levels was found between CBM with and CBM without metastasis (P > 0.05). The results were comparative to those of Northern blotting of the same cases. The authors concluded that, as also in the study of Northern blotting, the expression of nm23-H1 mRNA significantly correlated inversely with lymph node metastasis in CBM, while the expression of nm23-H2 mRNA not. Q-RT-PCR was a useful method to detect the mRNA levels of nm23 gene in buccal carcinoma. PMID- 11480036 TI - [An experimental study of the growth pattern and mechanisms of surgically induced cleft palate and palatoplasty on maxillary growth in dogs]. AB - The authors observed the changing process of length, width and height of maxillary growth in experimental animals, the experimental animals were divided into the unoperated control group, the surgical induced cleft palate group, the repaired cleft palate group and the group of implanted autograft bone in total hard palate. The method of a serial dorsoventral and lateral cephalograms were used in the study. The experimental result showed: the rate of maxillary growth began to change from two to four weeks and finished in the eighteenth week after operation. There was a remarkable different rate of maxillary growth among the four groups. The track of premaxillary growth had a difference between unoperated control group and the others. The experimental result suggested that the direction of growth in nasal septal cartilage and the conduction of force from premaxillary to maxillary were inhibited. It is a main reason for maxillary growth disturbance of the cleft palate who had or had not surgery. PMID- 11480037 TI - [An experimental study of the influence and mechanism of surgically induced cleft palate and palatoplasty on teeth and dental arch in dogs]. AB - The dogs were selected as the experimental subjects in this study. The authors continuously observed the maxillary teeth and dental arch form in preoperation and 2, 4, 6, 10, 14, 18, 22, 26, 30, 34 weeks of post operation to all the animals. 40 puppies were divided into four groups, which were unoperated control group, surgically induced cleft palate group, cleft palate repaired group and cleft palate implanted bone group. The experimental result showed: there were remarkable differences at rate of itself growth between deciduous dentition and permanent dentition in the four groups. The result suggested that the cleft palate was a reason for disturbances of dental arch development, an exposed bone wound in hard palate of maxillary after palatoplasty was an important reason that it made maxillary dental arch growth problem heavier, authors could prevent the disturbs the transverse development of the maxillary dental arch from cleft palate who had or had not repaired by implanted bone in total cleft palate. PMID- 11480038 TI - [IGF-I mRNA expression level of cell in the interfaces of bone and four kinds of substitutive materials for bone tissues by the technique in situ hybridization]. AB - Four kinds of substitutive materials for hard tissues were inserted into tibias of rat, effect of substitutive materials on IGF-I mRNA expression level of cell in the interfaces of bone and four kinds of substitutive materials were studied by the technique in situ hybridization. It showed that the biomaterials could affect the level of IGF-I mRNA expression, the enhanced expression level was associated with cell proliferation, formation of matrix and calcification, formation of new bone. Measurement of IGF-I mRNA could attribute to judge the functional state of cell, and to predict the final character of the interface. PMID- 11480039 TI - [The research on effect of four kinds of substitutive materials for the bone tissues on IGF-I mRNA expression level of osteoblast]. AB - The authors studied the expression level of IGF-I mRNA of osteoblasts cultured from the rats' calvarias, affected by four kinds of substitutive materials for bone tissues, using the technique in situ hybridization. The result showed that the substitutive materials could affect the expression level of IGF-I mRNA of osteoblasts. The measurement of expression regulation may be a parameter for evaluation of biocompatibility of substitutive materials for bone tissues. PMID- 11480040 TI - [An experimental study of morphologic and element analyses at the interface between bone and substitutive materials for human bone tissues]. AB - The implant materials chosen in this study included hydroxy-poly (calcium natrium phosphate), hydroxyapatite, bioglass ceramic and Ti-6Al-4V. The implants were inserted into the mandibles of dogs. The samples from dogs were regularly observed by X-ray examination, optical microscope, scanning electron microscope and X-ray energy spectrograph. The results showed that all the implants had integrated into the surrounding bone tissue by the third month. During the early stage of the experiment, there were a lot of S, Si at the bone-implant interface. After new bone became mature, the contents of Ca, P increased, but the contents of S, Si decreased at the interface. PMID- 11480041 TI - [A study on bonding performance of DPR dental adhesive]. AB - Phosphoric acid esters can be used as restorative materials for human teeth and bones. The authors investigated bonding performance of phosphoric acid diester (DPR) with tensile bond test. The results showed: (1) DPR dental adhesive could markedly improve the tensile bond strength (TBS) to human enamel, dentin, dental nonprecious metals and self-curing denture plastics, it is used widely; (2) Three kinds of the etchants could also improve markedly TBS. 37% phosphoric acid and DT I were respectively applicable to enamel and dentin. DT-II was available to both of them. PMID- 11480042 TI - [Experimental study on aqueous areca nut extracts inducing oral submucous fibrosis in rats. I. Observation of histomorphology]. AB - The purpose of this study was to establish the animal model of oral submucous fibrosis (OSF) in buccal mucosa of SD rats by submucosa injecting and surface painting aqueous areca nut extracts (AANE). The animals of all groups were killed randomly in the 12th, 16th, 22nd and 28th week. The buccal mucosae were collected, and were detected by light microscope and transmission electron microscope. The results were obtained: at different stages, the OSF-like lesions appeared in different degrees among some animals of the model groups, including atrophic epithelium with loss or partial loss of rate pegs, inflammation, accumulation and disorder of collagen fibre in juxta-epithelial region, along with concomitant muscle degeneration. The lesions didn't reverse obviously themselves after the authors stopped using AANE. Histomorphologic changes didn't appear in any animal of the control groups. The results indicated that the typical and stable OSF animal model were induced rapidly by submucosa injecting and surface painting AANE. PMID- 11480043 TI - [Experimental study on aqueous areca nut extracts inducing oral submucous fibrosis in rats. II. Effection of mast cells on collagen metabolism]. AB - The effection of mast cells (MC) on collagen metabolism of oral submucous fibrosis (OSF) induced by aqueous areca nut extracts (AANE) in rats was studied by transmission electron microscope, histochemical and biochemical analyses. The results revealed that there was a close relationship between MC and fibroblats (FB) in the pathogensis of OSF; MC which appeared in the buccal mucosa in the model groups increased significantly and became more active in the function. A great deal of collagen compacted in the buccal mucosa in the model groups. The contents of tissue hydroxyproline (Hyp) in the model groups were obviously higher than those in the control groups. Moreover, the longer the time of irritation continued, the higher the contents were. The number of MC had a higher positive correlation with the contents of tissue Hyp at the same time. Total serum Hyp had no significant difference among all the groups and any stages. The results indicated that some aqueous components of areca nuts might disturb collagen metabolism by accumulation and activation of MC. PMID- 11480044 TI - [A comparative study on the blood vessel sclerosing effect of pingyungmycin and Natrii morrhuates]. AB - Twelve adult rabbits were used in this experiment. After posterion auricular veins were temporarily blocked, pingyungmycin and natrii morrhuatis were injected separately and their effect on the blood vessels was studied. The results showed that pingyungmycin caused endotheliac cell oedema, degeneration, vascular shrinkage, vascular wall thickening, while the effect of natrii morrhuatis on the blood vessels was thrombosis, organization of thrombus and closure of the lumen of the blood vessels. The sclerosing mechanisms of the two medicines and their clinical applications were discussed. PMID- 11480045 TI - [The study on apatite castable glass-ceramics in composition and physicochemical properties]. AB - The authors reported the result of the study of the glass formation range and physicochemical characteristics of apatite (APA) castable ceramics in the CaO-MgO Al2O3-SiO2-P2O5 system. The relationship between its component and casting properties, the mechanical property and chemical stability was mainly discussed. The result showed that homogeneous glass could be formed in the experimental scope. This glass crystallized enamel-analogous apatite phase. It has improved mechanical property of parent glass, the bending strength was up to 165 MPa and possessed a fine chemical stability. This investigation could provide a new castable ceramics material for dental restoration. PMID- 11480046 TI - [The influence of basement membrane matrix on the attachment of human gingival epithelial cell to titanium: a scanning electron microscopic study]. AB - In order to investigate the factors which could enhance the integrity of the biological seal between implant and epithelial cells, basement membrane matrigel was coated on the surface of smooth pure titanium. SEM observations showed that the spreading of primary cultured human gingival epithelial cells on Matrigel coated titanium surface was better than that of control groups on the third and the sixth day. This result suggests that the attachment of human gingival epithelial cells to titanium can be enhanced by the basement membrane matrigel. PMID- 11480047 TI - [The experimental observation on characteristics of soft tissues infection in maxillofacial region wounded by high velocity missile]. AB - In this experimental study, maxillofacial regions of dogs were wounded by steel spheres, which weight 1.03 g at a mean primary velocity of 1500 m/s. Aerobe and anaerobe bacteria in the muscles of different interval from wound track edge were cultured at different time after wound. The results showed that there was bacteria infection in the soft tissues between 0 and 0.5 cm from wound track edge in 6 hours after wound. The quantity of bacteria obviously increased following time prolonging, on the contrary, it was much lower than threshold value of bacteria infection within 24 hours. The experimental results suggested that there was obvious bacteria infection in the soft tissues within 0.5 cm distance from wound track edge in maxillofacial firearm wound region. During the debridement removing soft tissues within 0.5 cm distance from wound track edge, the infection of wound region could be controlled. Using effective antibiotics is an important measure of controlling infection of wound after debridement. PMID- 11480048 TI - [A study on the frequency of sister chromatid exchanges in patients with oral submucous fibrosis in peripheral blood lymphocytes]. AB - The frequency of sister chromatid exchanges (SCEs) in peripheral blood lymphocytes in the patients (n = 27) with oral submucous fibrosis (OSF) with the habit of chewing areca nuts were determined by BrdU-Giemsa staining. The healthy persons (n = 14) with the same habit and the normal controls without the habit (n = 44) were studied at the same time. The results showed that the frequency of SCEs in the first two groups were obviously higher than that in the last one (P < 0.001). The frequency of SCEs in the patients were also higher than that of the healthy persons with the habit (P < 0.001). These suggest that there are some substances in the areca nuts which can induce the mutation and/or malignant transformation of cells. The habit of chewing areca nuts might seriously disturb the stability of chromosome. OSF is a precancer condition. The occurrence of it perhaps has some hereditary background or genetic susceptibility. PMID- 11480049 TI - [The expression of IL-1 alpha gene in gingival tissue]. AB - IL-1 is secreted by a variety of cells and plays an important role in the network of cytokines. In this study, the expression of IL-1 alpha in gingival tissue was determined with reverse transcription-polymerase chain reaction (RT-PCR). The PCR product was confirmed by DNA sequencing. The results showed that IL-1 alpha transcription was detected in all 5 inflammatory gingival tissue samples, but it was not delected in the normal gingival tissue samples and in the samples after treatment. These findings suggested that IL-1 alpha may be a specific indicator of periodontal disease, and a potential mediator in pathogenesis of periodontitis. PMID- 11480050 TI - [The study on SDS-PAGE technic classification of the whole protein of oral bacteria]. AB - Oral flora was classificated by SDS-PAGE combined with Comassie blue staining and silver staining. The results showed that each species of the test oral bacteria expressed its specific electrophorectic protein pattern, the way of sample preparation, staining did not change the protein pattern. This indicated that SDS PAGE could be used to classificate oral flora. The protein pattern was more obvious among the genus than that of within the genus. This showed that different relationship was among the bacteria. PMID- 11480051 TI - [Research on the relation between the biting force with complete denture and compression of the mandibular denture bearing mucosa]. AB - This experiment was designed to study the relation between the biting force and compression mucosal recovery and recovering time of the denture bearing mucosa with ten edentulous patients by the occlusometer and the photoelectric movement transducer. The results showed that the favourable threshold of biting force for denture bearing mucosa was 1.3 kg in anterior tooth region and 2.5 kg in molar region, up to 30 minutes after removal of biting force, the mucosal thickness was still 0.20 mm in anterior tooth region and 0.11 mm in molar region. According to these characteristics of the denture bearing mucosal compression and recovery, masticatory force of the complete dentures should be limited below above mentioned threshold. PMID- 11480052 TI - [A study of therapeutic efficacy of Koukangning gargle in the treatment of oral mucositis]. AB - A study of therapeutic efficacy was carried out in 118 patients with oral mucositis (recurrent oral aphthous ulceration, erosive lichen planus, herptic stomatitis and infectious oral mucosal hematoma) at random. As a topical medication, Koukangning gargle was applied to 59 patients of the experimental group, 29 patients of the control group 1 applied Dobell's solution to rinse their mouths, and 30 patients of the control group 2 applied Koutai solution. A satisfactory result was observed in the experimental group, its effectiveness and effective rates rose more than those of control group 1 at both treatments for 3 days and 6 days. Between the two groups, there were markedly significant difference (P < 0.005, P < 0.01, P < 0.005, P < 0.005). Koukangning gargle was nontoxic and no side-effects in the clinical tests. Its efficacy was no significant difference compared with the control group 2. Experimental study demonstrates that it possesses anti-inflamation effect and excellent antibiotic effect, therefore it is a new gargle for rinse mouth combined traditional Chinese medicine with Western medicine. PMID- 11480053 TI - [Experimental study of artificial bone composite with high biological induction in repairing bone defect]. AB - On the basis of the animal model of segmental ultra defects in rabbits, two different composite artificial bones (HA-bBMP-Co and HA-Co) were first used to repair the segmental bone defect respectively in 12 rabbits. The results of X ray, gross, stereomicroscopic and histological examination were found that HA bBMP-Co was much faster and much better than HA-Co, involving induction of new bone formation in quality and quantity. It is suggested that there is a widespread prospect in use of HA-bBMP-Co as a bone substitute material. PMID- 11480054 TI - [Orthodontic treatment for the surgical correction of mandibular skeletal retrognathism]. AB - The purpose of this paper is to discuss the combining orthodontic treatment that is based on seventeen consecutive patients of severe mandibular skeletal retrognathism who have been treated by orthognathic surgery. Not only the presurgical preparation, but also the presurgical and postsurgical orthodontic treatment and the use of interocclusal splints were introduced. The goals of orthodontic treatment for patients with orthognathic surgery are (1) decompensating the teeth, expanding the upper dental arch, (2) aligning the teeth, (3) leveling the curve of Spee, (4) making the shape and width of upper and lower dental arch harmonious, (5) getting the occlusal equilibrium and fine occlusal contact. The author's experience indicates that: in order to get satisfactory results of esthetics and function, the combination of surgical and orthodontic treatment, the presurgical and postsurgical orthodontic treatment is absolutely necessary and important. PMID- 11480055 TI - [The effect of deture relining on the electromyogram of masticatory muscles in complete denture wearers]. AB - The aim of present study is to assess the effect of complete denture relining on the electromyogram of masticatory muscles. The electromyography and the computer EMG system were used for collecting and processing bilateral masseter and anterior temporal muscles activities before and after denture relining. The results showed that the mean myoelectric potentials of the masticatory muscles was increased during chewing and maximal biting when the dentures were relined. The conclusion was that the level of the masticatory muscle activity was increased when a clinically acceptable occlusal vertical dimension was obtained by using relining method. PMID- 11480056 TI - [Matched case control study for investigating of the association between psychosocial factors and TMJDS]. AB - A 1:1 matched case control study was carried out in 81 cases with temporomandibular joint dysfunction syndrome (TMJDS) who came to a TMJ clinic of a tertiary dental centre for help and 81 controls without TMJDS to investigate the association between psychosocial factors and TMJDS. The results showed that anxiety and depression might be one of the risk factors of TMJDS. Trait anxiety and Eysenck's neuroticism might be the inherent personality factor of anxiety, and the later might be the basis of depression. Life events and social support might be the external factors of TMJDS. PMID- 11480057 TI - [Anesthetic management to reduce blood loose in surgical process of palatoplasty]. AB - 869 cases with palate cleft, aged from 2 to 12 years old, received palatoplasty surgery in Department of Oral Maxillofacial Surgery, West China University of Medical Science from 1987 to 1996 were reviewed. The effect of anesthetic method on blood loose during surgical procedure was analyzed specifically. It was found that the amount of blood loose was quite different with different anesthetic method. Simple method of anesthetic medication which could dramatically reduce the amount of blood loose in palatoplasty operation was recommended. The key points of anesthetic method for palatoplasty operation, especially in application for children were discussed. PMID- 11480058 TI - [Maxillofacial injury: clinical analysis of 284 cases]. AB - Maxillofacial injury is common in war or in the routine life. Always accompanying trauma of other sites of the body, it has its own characters and leads to serious outcome. This article made a clinical analysis of 284 cases of maxillofacial injury from following aspects: onset of wounded, characters and chief complications of maxillofacial soft and hard tissues injury and foreign bodies left in place. The authors also discussed their experiences at emergency management. PMID- 11480059 TI - [Conjugation and identification of the surface protein antigen P1 from streptococcus mutans MT6R with the cholera toxin B subunit and procholeragenoid]. AB - To make an effective antigen to prevent dental caries, the surface protein antigen P1(Ag I/II) was purified and identified by the rabbit anti-Ag I/II serum and was covalently conjugated to CTB and PCG by SPDP respectively. GM1-ELISA showed that both the conjugated P1-CTB and P1-PCG possess the ability to bind GM1 specifically without losing the antigenicity of P1. This results showed that the conjugated antigens could be used in the immune prevention against dental caries. PMID- 11480060 TI - [A primary observation of the active immune prevention effects against dental caries using the streptococcus mutans surface protein P1 in rats]. AB - To observe the anti-caries effects of a small dose of the surface protein antigen P1. The SD rats were immunized with P1-CTB and P1-PCG through IN (intranasal immunization) and IG (intragastric immunization). The results showed that caries was reduced heavily. There was no obvious difference between IN and IG. P1-CTB and P1-PCG can be used as an antigen to prevent the dental caries, but much further research work still need to do. PMID- 11480061 TI - [An animal model for the mucosal immunization with the surface protein antigen P1 from streptococcus mutans and immune enhancement of cholera toxin]. AB - To study the immune enhancement of cholera toxin by developing an mucosal immune animal model, SD rats were immunized using different antigen composition through intragastric route (I.G). The results showed that the effects of immunization became stronger after P1 was conjugated to cholera toxin B subunit (CTB) or procholeragenoid (PCG). Even microgram dose of P1 could induce the rising of anti P1 SIgA antibodies in the rat's saliva, but the antibodies risied little without this adjuvant. These implayed that the conjugation was much necessary in IG immunization. The immune enhancement of PCG was greater than that of CTB. PMID- 11480062 TI - [The study of actinomyces viscosus adhere to teeth. III. The identification of mutants of A. viscosus and preparation of rabbit IgG against type 1 and type 2 fimbriae]. AB - The authors prepared rabbit antisera against 5519 and 5951. The IgGs against type 1, type 2 fimbriae were purified from the antisera using sera absorbed with the other strain and IgG purification method. It was identified by SDS-PAGE and ELISA. This result suggested the purified proteins were the IgG against type 1 and type 2 fimbriae. PMID- 11480063 TI - [The study of actinomyces viscosus adhere to teeth. IV. The study of salivary protein receptors of A. viscosus]. AB - In order to determine the receptors of A. viscosus for adhesion, 7 kinds of main purified proteins from pellicle formed by whole saliva were tested. The result indicated that: 1. APRP, statherin, PRPG and SIgA were the receptors of standard and fresh isolated A. viscosus; 2. APRP and stather promoted the adherence of 5519 (1+2-) to HA, and SIgA promoted the adherence 5951 (1-2+) to HA; PRPG was the common receptor of the two fimbriae. It suggested that there were receptors not only type 1 fimbriae but also type 2 in pellicle. Two types of fimbriae were involved in the adherence of A. viscosus to pellicle. The difference existed in the receptor of adherence and the activity site of the same receptor. PMID- 11480064 TI - [Ultramicrostructure study of artificial bone composite with high biological induction in repairing bone defect]. AB - The scanning electric microscopy (SEM). X-ray energy graph and Ca, P contents analysis were adopted in studying the mechanism and superiority of two different composite artificial bones (HA-bBMP-Co and HA-Co) to repair segmental bone defect in rabbits. The results indicated that HA-bBMP-Co had a double effects of bone induction and bone conduction. No difference of P content in two weeks after reconstructive surgery was found, but in other periods, there was an obvious difference in Ca and P content between HA-bBMP-Co group and HA-Co group (P < 0.05). PMID- 11480065 TI - [A three-dimensional finite element stress analysis of implant-supported prosthesis and its supporting tissue in the edentulous mandible. Part 5. The influence of implant type in complete implant overdenture and its supporting tissue]. AB - The stress distribution of implant, implant-bone interface, alveolar ridge surface and the overdenture when supported by cylindrical-type or blade-type implants under three kinds of occlusion, were investigated respectively in this study. The results showed that the extreme principle stress was greater when the denture was supported by blade-type implants than by cylindrical type one. The stress distribution of the system with application of cylindrical type implant is more acceptable. PMID- 11480066 TI - [A three-dimensional finite element stress analysis of implant-supported prosthesis and its supporting tissue in the edentulous mandible. Part 6. The influence of superstructure in mandibular complete implant overdenture and its supporting tissue]. AB - The effect of superstructure, including telescope, conventional-bar and modified bar, on the stress distribution of mandibular complete implant overdenture and its supporting tissue was investigated in this study. The results demonstrated that bar could lower the extreme principle stress of the implant, but the stress distribution of implant-bone interface was better when the implants were not linked with each other. Modified-bar is not preferred because it increases the stress of implant and its interface. PMID- 11480067 TI - [Reinnervation of pectoralis major myoflap by the hypoglossal nerve: an experimental study in rabbits]. AB - In order to investigate the morphological, electrophysiological and histochemical characteristics of the pectoralis major myoflaps with original nerve innervation and with hypoglossal nerve reinnervation, after denervation, the model of myoflap transposition to reconstruct the tongue was adopted on 27 New Zealand white rabbits. After the myoflaps were transferred, original nerve innervation (ON), hypoglossal-genioglossus muscle branch nerve implantation (NI) and hypoglossal genioglossus muscle neuromuscular pedicle implantation (NMP) were performed. The flaps were examined at the intervals of 4, 8, 12 weeks and 16 weeks after operation. The results showed that the denervated myoflaps were reinnervated in NI and NMP, myoflaps contracted when the hypoglossal nerves were stimulated. The histochemical properties of muscle fibers changed. Some type II fibers in the myoflaps transformed into type I fibers, but the pattern of distribution of the muscle fibers displayed mosaic and was similar to that of genioglossus muscle. In terms of speed and scope of reinnervation, the NMP was superior to the NI. PMID- 11480068 TI - [A research on rat osteoblasts culture in vitro]. AB - Osteoblasts were isolated from Sprague Dawley rats of 1-3 days on the frontal parietal bones and were confirmed by testing for multiple biological features. The results showed that rat osteoblasts in vitro had the same morphological feature, ALP activity, the function of synthetic BMP as in vivo and it had the ability of calcification in vitro. The results proved that these rat osteoblasts in vitro had the same function and features as in vivo and it could be used to study cytocompatibility on replacement materials of human body's hard tissues and biomedical to make experiments related to bone tissues. PMID- 11480069 TI - [The study and primary application of multimedia teaching and learning system for diseases of temporomandibular joint]. AB - A multimedia computer-assistant teaching and learning system for diseases of temporomandibular joint was reported in this paper. It was designed and implemented in Windows (Chinese) on PC 486. It was emphasized that interactive teaching and learning was an available, funny method and student was the focus of teaching and learning in this system. A satisfied result was obtained by utilizing of this multimedia teaching and learning system for primary application. PMID- 11480070 TI - [Applications of computer image processing and graph processing technologies in digital modelling of 3-D shapes of teeth, jaw and prosthesis]. AB - In this paper, the images of sliced sections or CT sections of the specimen of teeth, jaw and prosthesis were processed by a computer image analysing system, and the plane shape of the 3-D objects of interest on the sections was converted into digital images using a special software module for contour analysing. Then the images were adjusted in scale, modified in coordinates and smoothed in shape, and the data format was transferred for finite-element analysis by a specially designed software. In such ways a method for the digital modelling of the 3-D shape of teeth, jaw and prosthesis was successfully established. PMID- 11480071 TI - [To study, propagate and apply the Chinese classification of tuberculosis]. PMID- 11480072 TI - [ Measuring and assessing the quality of life of patients with pulmonary tuberculosis]. AB - OBJECTIVE: To measure and assess the quality of life (QoL) of patients with pulmonary tuberculosis. METHOD: QoL of 228 patients with pulmonary tuberculosis and 228 healty controls were marked by MOS SF-36, QLI and KPS scales, and single and multiple stepwise regression analysis were made to evaluate factors affecting QoL. RESULT: The average total mark of SF-36, marks of physical functioning, role physical, mental health, role-emotional, social functioning, vitality, bodily pain and general health were respectively 57 +/- 17, 65 +/- 28, 22 +/- 32, 61 +/ 20, 30 +/- 36, 54 +/- 27, 56 +/- 21, 65 +/- 24 and 53 +/- 15 in the pulmonary tuberculosis group, while 77 +/- 8, 84 +/- 23, 81 +/- 34, 75 +/- 18, 81 +/- 33, 83 +/- 23, 61 +/- 21, 75 +/- 24 and 72 +/- 20 in the controls. Statistically significant differences were found between the two groups (all P values < 0.01). The average marks of QLI and KPS were 7.4 +/- 2.0 and 77 +/- 17 respectively in the pulmonary tuberculosis group, and significant differences were also found comparing with the controls (9.6 +/- 0.8, 97 +/- 9) (P < 0.01). There were statistically significant correlation among the total mark of SF-36 and the marks of the above 8 subdividions in patients with pulmonary tuberculosis. The correlation coefficients between marks of SF-36 and QLI, KPS were 0.7841, 0.8931 respectively (P < 0.001). The factors affecting the marks of SF-36 of the patients were focus size of infection, counts of white blood cells, complications, elevated ALT and duration of disease. CONCLUSION: The SF-36 scale is suitable for measurement of QoL of patients with pulmonary tuberculosis. The QoL of patients with pulmonary tuberculosis declines. The main factors affecting the QoL of the patients are focus size of infection, counts of white blood cells, complications, elevated ALT and duration of disease. PMID- 11480073 TI - [Analysis on quality control of drug susceptibility testing in the surveillance of drug resistance in tuberculosis in Henan]. AB - OBJECTIVE: To evaluate the working quality of Henan province reference laboratory(PRL) in the surveillance of drug resistance (DRS) in Henan. METHOD: West Pacific Regional (WPR) Supranation Reference Laboratory(SRL) in South Korea conducted three times of quality control of drug susceptibility testing (DST) from SRL to PRL and to national reference laboratories(NRLs) in the region at the same time, and once of which from PRL to SRL. The exchanging strains of M. tuberculosis, which were clinical isolated ones containing both drug susceptible and drug resistant cultures, were tested for resistance to SM, INH, RFP and EMB. The DST procedure was proportion method, and L-J medium was used. RESULT: It was shown that the sensitivities for most of the four drugs in the three times of DST quality control from SRL to PRL were higher than their specificities, and that the sensitivities for both INH and RFP were 100%. And the accuracy, or agreement, for both INH and RFP was more than 90%, except for that of RFP in the first time. Compared with the results from SRL to NRLs (including Henan PRL), except for the specificity for SM in WPR being higher than that in Henan (84% versus 70%, P < 0.05), there was no significant difference between Henan and WPR on the specificity and sensitivity of the other drugs. Similar characteristics can be found in the results from PRL to SRL. And the efficiency for SM, INH and RFP was all higher than 90%. CONCLUSION: The quality control on Henan PRL meet the requirement of the WHO/IUATLD guidelines for surveillance of drug resistance in tuberculosis(1997). PMID- 11480074 TI - [Study on the mutations of ahpC genes in M. tuberculosis isoniazid-resistant isolates]. AB - OBJECTIVE: To investigate the mutations of ahpC genes in M. tuberculosis isoniazid-resistant isolates, and to study the correlation between ahpC alterations and isoniazid resistance. METHOD: Analyzing the ahpC coding sequences and ahpC promoters in 62 M. tuberculosis clinical isolates with PCR-SSCP method. M. tuberculosis strain H37RV was used as control. RESULT: The ahpC coding sequences showed normal SSCP profiles in 62 clinical isolates. AhpC promoter mutations were identified in 5 of 12 high isoniazid-resistant strains, and were not altered in 18 low isoniazid-resistant and 32 drug-sensitive isolates. CONCLUSION: The mutation of ahpC coding gene was not correlated with isoniazid resistant M. tuberculosis isolates. AhpC promoter mutation was a compensatory change in katG-defective and catalase-negative isolates, may be used as a indirect marker for high resistance to isoniazid. PMID- 11480075 TI - [Determination of telomerase activity in BALF cells from lung cancer patients]. AB - OBJECTIVE: To investigate the telomerase activity in BALF cells from patients with lung cancer. METHOD: The silver staining and PCR technique were used to detect the telomerase activity in BALF cells from 30 patients with lung cancer and 30 patients with chronic bronchitis or benign pulmonary diseases. RESULT: The telomerase activity in BALF cells was positive in all the patients with lung cancer (25/30, 83%), and in none of the patients with chronic bronchitis or benign pulmonary diseases (0/30, 0%). The difference was highly significant (P < 0.001). In the same patient with lung cancer, the positive rate of telomerase activity was 83% in BALF cells from the diseased lung, but only 27% in BALF cells from the normal lung (P < 0.001). CONCLUSION: The telomerase activity may be used as a diagnostic marker for lung cancer. There is a correlation between telomerase activity and the development of lung cancer. PMID- 11480076 TI - [Expression of epidermal growth factor receptor (EGFR), platelet growth factor receptor (PGFR) and insulin growth factor-1 receptor (IGF-1R) on lung cancer cell]. AB - OBJECTIVE: To evaluate the expression of growth factor receptors (GFRs) on lung cancer cells and the relation between the expression of GFRs and metastasis of lung cancer. METHOD: The expression of EGFR, PDGFR and IGF-1R was detected with immunohistochemical method using monoclonal antibodies. RESULT: The expression of EGFR, PDGFR and IGF-1R showed higher positive rates in lung cancer cells than those in normal lung tissues, and the expression was increased as the disease was advanced. CONCLUSION: The increased expression of EGFR, PDGFR and IGF-1R is related with the metastasis of lung cancer. PMID- 11480077 TI - [Study on gas exchange of the patients with lung cancer in perioperation]. AB - OBJECTIVE: To study changes of gas exchange in perioperative patients with lung cancer. METHOD: Indices including P/F, a/A, P50O2 and PaCO2 on gas exchange in 68 male patients(50-78 y) with lung cancer were monitored on preoperative day 1 2 3 and postoperative day 7. RESULT: Comparing with preoperation data P/F a/A decreased significantly in 3 days(P < 0.01) and PaCO2 P50O2 increased significantly on the day after operation (P < 0.01), especially in the group of pneumonectomy and abnormal ventilation, hypoxemia measured in preoperation. CONCLUSION: The gas exchange of the patients with lung cancer reduced significantly in postoperative 3 days. PMID- 11480078 TI - [Pulmonary lymphangitic carcinomatosis]. AB - OBJECTIVE: To explore the clinical manifestations of pulmonary lymphangitic carcinomatosis (PLC), to analyse its associated diagnostic methods, and to improve the understanding of PLC and its diagnosis. METHOD: Retrospective analysis of 4 cases of PLC and review of the literature. RESULT: The clinical manifestations of PLC include: (1) dyspnea and cough; (2) normal or restrictive pattern ventilation; (3) diffuse or local reticulonodular infiltrates in the lung like interstitial fibrosis and pleural effusion on chest radiograph; (4) CT and high-resolution CT (HRCT) scans reveal a beaded chain appearance caused by uneven thickening of the interlobular septa and pleural membrane, polygonal thickening of bronchovascular bundles, and mediastinal lymphadenopathy as well. CONCLUSION: These clinical data suggest that any manifestations similar to pulmonary interstitial fibrosis complicated with pleural effusion and paratracheal lymphadenopathy should be further differentiated from PLC by HRCT and pleural lung tissue biopsy. PMID- 11480079 TI - [Preliminary observation of immunotherapy on asthmatic guinea pig]. AB - OBJECTIVE: To observe the effect of the anti-IL-5 monoclonal antibody(McAb) on eosinophils of the guinea pigs with asthma. METHOD: Sensitized guinea pigs by OVA were divided into two groups randomly, which were treated with the McAb and normal saline (NS), respectively. Difference of peripheral blood eosinophil count (PBEC) among basal pre-treatment and post-treatment, and the difference of bronchoalveolar lavage fluid eosinophil count (BALFEC) between two groups above were compared. RESULT: The PBEC after treatment with the McAb [(1.63 +/- 0.26) x 10(9)/L] was lower than that before treatment [(1.76 +/- 0.27) x 10(9)/L] (P < 0.05) and the BALFEC of treatment group with the McAb was lower than that with NS [(0.41 +/- 0.06) x 10(9)/L vs. (0.46 +/- 0.07) x 10(9)/L] (P < 0.05). CONCLUSION: The anti-IL-5 McAb has the ability to reduce the PBEC and the BALFEC. PMID- 11480080 TI - [The regulating effect of protein kinase C pathway on the airway tone]. AB - OBJECTIVE: To investigate if the role of protein kinase C(PKC) signal pathway in the regulation of tracheal and lung parenchymal strips tone in normal guinea-pigs is different from that in ovalbumin-sensitized ones. METHOD: By measuring the tones of tracheal and lung parenchymal strips isolated from normal and ovalbumin sensitized guinea-pigs, the effects of PKC-specific activator PMA and inhibitor Ro31-8220 were observed and compared. RESULT: (1) PMA caused concentration dependent relaxations in the tracheal strips of both normal (n = 7) and sensitized animals(n = 7) and the relaxant effects were significantly stronger on the strips from the sensitized animals than on those from the normal ones(P < 0.05). The relaxing effects could be completely inhibited by Ro31-8220. (2) PMA led to concentration-dependent contractions in the lung parenchymal strips and the contractile effects were significantly stronger on the strips from the sensitized animals (n = 5) than on those(n = 6) from the normal ones (P < 0.05). This contractile response could be abolished by Ro31-8220. (3) Ro31-8220 could partly suppress the contractile response of sensitized lung parenchymal strips to in vitro antigen attacks (n = 10, P < 0.01). CONCLUSION: (1) The regulating effects of PKC on airway smooth muscle tone differ in different locations of the guinea-pig airways. (2) The function of PKC signal pathway in airway smooth muscle from sensitized guinea-pig is increased compared with that from normal one. It is suggested that the role of PKC signal pathway may be important in the pathogenesis of asthma. PMID- 11480081 TI - [Analysis of basic data of the study on prevention and treatment of COPD and chronic cor pulmonale]. AB - OBJECTIVE: The basic data were analysed to carry out the project of "the study on prevention and treatment of COPD and chronic cor pulmonale population" and to provide some scientific basis for making strategies of prevention and treatment of COPD and chronic cor pulmonale in communities. METHOD: The basic survey of the study was carried out on 102,230 rural people in the spring of 1992. At first the family inquiry into the chronic airway inflammation history and smoking index > or = 300 were made in population aged 15 years or more. Those with FEV1/FVC < 70% in lung function were considered as having COPD, and chronic cor pulmonale was diagnosed according to national criteria by chest radiography and electrocardiography. RESULT: There were 67,251 people aged > or = 15 years in 102,230 population, among them 33,119 were male and 34,132 were female. Of the 7,400 subjects who should be investigated, 6,536 subjects were really investigated (88.3%), which accounted for 9.7% of > or = 15 year population. 2,020 people were diagnosed as having COPD, the prevalence of COPD being 3.0% in total population(> or = 15 years), and the highest was 4.5% in Beijing. The prevalences of COPD in smokers, subjects with chronic airway inflammation history and both of them were 24.6%, 34.7% and 40.4%, respectively. The sole smoking accounted for 40.7% and chronic airway inflammation history for 28.4% in the all COPD causes, respectively. 71.6% of COPD were related to smoking. 452 patients were diagnosed as having chronic cor pulmonale, which accounted for 23.1% of COPD, for 6.7@1000 of > or = 15 year population, for 4.4@1000 in total population. CONCLUSION: The basic data demonstrate that the prevalence of COPD and chronic cor pulmonale are quite high. There are remarkable differences of the prevalences among different regions. The primary causes of COPD and chronic cor pulmonale are smoking and chronic airway inflammation. The strategy of prevention and treatment of them should be a comprehensive one, the key measure be smoking cessation and controling airway inflammation. PMID- 11480082 TI - [Gene expression of messenger RNA coding for procollagen and tumour necrosis factor-alpha in lungs undergoing bleomycin-induced pulmonary fibrosis]. AB - OBJECTIVE: mRNA for tumour necrosis factor-alpha(TNF-alpha) and procollagen I [pro alpha 1(I)] and III (pro alpha 1(III) was measured in bleomycin treated mice to evaluate their roles in pulmonary fibrosis. METHOD: 30 male Wistar rats were divided into normal control group and groups treated by bleomycin on days 3, 7, 14, 21, 30. Each group included 5 rats, and the rats were treated by single intratracheal instillation with sterile saline solution or with 5 mg/kg body weight of bleomycin. The rats were sacrificed at different days, and total RNA from the lungs of bleomycin or saline solution instilled rats was extracted. Expression of TNF-alpha, pro alpha 1 (I) and pro alpha 1 (III) mRNA was measured by dot blot and Northern blot hybridization. RESULT: The levels of TNF-alpha mRNA in lungs of bleomycin groups on days 3,7 were significantly higher than those of control group (t = 10.33 and 12.54 respectively; P < 0.01). The levels of pro alpha 1 (I) and pro alpha 1(III) mRNA in lungs of bleomycin group on day 7 were higher than those of control group(t = 10.56 and 24.23 respectively; P < 0.01), After reaching the peak levels in two weeks, the levels of pro alpha 1(III) mRNA in lungs of bleomycin group on day 30 declined slowly to approximately the values of normal control group (t = 1.95; P > 0.05); but the levels of pro alpha 1(I) mRAN on day 30 after bleomycin administration were higher than those of control group (t = 16.67; P < 0.01). The mRNA coding pro alpha 1(I) and pro alpha 1(III) in lungs of rats had two mRNA species. CONCLUSION: The results suggested that the metabolism of mRNAs for pro alpha 1 (I) and pro alpha 1 (III) may be preferentially perturbed, and that increase of collagen gene expression in the transcriptional level might be involved in the mechanism of collagen accumulation. The increased expression of TNF-alpha gene may play an important role in the early events of bleomycin-induced pulmonary fibrosis. PMID- 11480083 TI - [Clinical study on the forced oscillatory pulmonary function test]. AB - OBJECTIVE: To discuss the change of airway resistance and phase angle (phi) in normal subjects, patients with COPD, asthma and pulmonary fibrosis when the new technique of the forced oscillatory pulmonary function test was applied to measure them as well as the change of pulmonary function of patients with asthma after using bronchodilator. METHOD: Use Custo-vitm forced oscillation device manufactured in Germany to determine airway resistance and phase angle and to compare their statistical differences between normal subjects and patients with pulmonary disease, and then comparing the improvement rates of their FEV1, F-V curve and airway resistance in patients with asthma after inhaling bricasol. RESULT: The measurements of group COPD and asthma were significantly higher than those of normal group (P < 0.05), and there were obvious frequency dependence in these two groups. The results of phase angle in each group of patients were significantly lower than those of normal group (P < 0.05), and the group of patients with pulmonary fibrosis only demonstrates a decrease in phase angle, and the improvement rate of airway resistance in patients with asthma after inhaling bricasol was significantly higher than FEV1 and F-V curve (P < 0.05). CONCLUSION: Forced oscillation is a convenient, quick, and noninvasive new method when applied to measure airway resistance and phase angle. It is especially suitable for patients with severe COPD and better than conventional pulmonary function test in observing therapeutic effect in asthma. PMID- 11480084 TI - [Advances in the chemoprophylaxis of tuberculosis]. PMID- 11480085 TI - [Shoulder endoprostheses--a special status in prosthesis implantation]. PMID- 11480086 TI - [Biomechanical considerations on shoulder joint prosthesis implantation]. AB - The biomechanical goals of prosthetic reconstruction of the shoulder are to restore the normal anatomy and range of motion, and to recreate the normal soft tissue balance of the static and dynamic stabilizers of the glenohumeral joint. An unconstrained prosthesis design best reproduces the physiological articulation and original anatomy of the shoulder. Humeral head components have been recently developed, which are adaptable to the variable anatomy of the proximal humerus (third generation design). A precise reconstruction of the three dimensional structure of the proximal humerus may lead to an improved functional outcome. However, there is still a lack of biomechanical data to support this concept. The optimal design of the glenoid component remains a challenge for future research. Specific issues including the choice of biomaterials, the optimum shape, radius of curvature, surface area of the articulation, component height and stem design remain under investigation. Although the prosthetic design represents an important factor in the success of glenohumeral arthroplasty, the surgical reconstruction of the soft tissues to recreate the normal soft tissue balance as well as postoperative rehabilitation determine the functional outcome. PMID- 11480087 TI - [Principles of shoulder prosthesis implantation]. AB - Patients with an intact rotator cuff and a humeral head that is centered in the glenoid fossa will benefit from both: a hemiarthroplasty and a total shoulder arthroplasty. However, the functional outcome following total shoulder arthroplasty is superior to that of hemiarthroplasty. Superior migration or mal positioning of the humeral head in the anterior or posterior direction are generally associated with a maximum active flexion of 90 degrees and a high rate of loosening of the glenoid component. Total shoulder arthroplasty leads to superior results in patients with osteoarthritis and mal-positioning of the humeral head in the posterior direction. However, if the head can not be centralized in the glenoid fossa a significant risk of glenoid loosening remains. A superior functional outcome of total shoulder arthroplasty in patients with rheumatoid arthritis can be observed. On the other hand inferior bone quality and a rotator cuff might lead to loosening of the glenoid component. Radiographic signs of glenoid loosening are frequently observed. However, these hardly require operative revisions. If a glenoid component can not be inserted, a bipolar or inverse prosthesis might be considered an alternative. PMID- 11480088 TI - [Glenohumeral joint. Anatomical aspects and implications for prosthesis design]. AB - Knowledge of normal anatomy and kinematics of the glenohumeral joint as well as analysis of the pathoanatomic changes that occur during the course of the disease is the condition for successful patient management. This review article critically discusses the most relevant geometrical variables. Systematically, normal anatomy, pathologic anatomy, and the implications for prosthetic design and implantation are presented. The most important aspects concerning the size and shape of the glenoid, the inclination of the articular surface in the horizontal and frontal planes, as well as the geometry of the scapular neck are discussed. The accumulated knowledge is of immediate practical use and might stimulate researchers and manufacturers to develop more adequate glenoid components. PMID- 11480089 TI - [Prosthesis implantation of the rheumatoid arthritis shoulder]. AB - Omarthritis occurs frequently during the early course of rheumatoid arthritis. Many rheumatoid patients ignore omarthritis because of the good compensation mechanism of the shoulder. Sonography and tomography enable early diagnosis of omarthritis before deterioration is radiologically visible. Arthroplasty gives better results when the musculotendinous rotator cuff is still intact. Therefore, arthroplasty should be performed before severe damage develops. Early indication for cup arthroplasty of the humeral head is justified because of better options in revision surgery. Glenoid components show a high incidence of radiolucency and loosening in rheumatoid patients. Cemented hemiarthroplasties show the lowest rate of loosening. PMID- 11480090 TI - [Endoprosthetic surface replacement of the head of the humerus]. AB - The concept of an endoprosthetic surface replacement of the humeral head differs from that of stemmed endoprostheses. It is the replacement of the destroyed joint surface with reconstruction of the normal anatomy and minimal bone resection. The aim of this prospective study was to evaluate the short-term results of a newly developed cup arthroplasty (Durom-Cup) for the humeral head. In a prospective study, 39 patients with 46 Durom-Cups were evaluated preoperatively and every 3 months postoperatively. The average follow-up was 15 +/- 9 months. The group included 28 shoulders with rheumatoid arthritis, 15 joints with osteoarthritis, and 3 humeral head necroses. The Constant-score and SAS-function score were used. The Constant-score increased from 20.25 +/- 9.06 points preoperatively to 46.62 +/- 14.05 at 3 months, to 48.11 +/- 14.49 at 6 months, and to 55.25 +/- 11.6 at 9 months postoperatively. The Constant-score stayed at this level during further follow-up and was 55.81 +/- 16.31 at 12 months postoperatively. The best results were seen in the group of humeral head necroses with a Constant-score of 71.0 +/- 12.2 compared to 54.66 +/- 13.89 in the group of osteoarthritis and 56.78 +/- 13.33 in patients with rheumatoid arthritis at 12 months postoperatively. The results with the Durom-Cup are encouraging so that cup arthroplasty seems to be a good alternative to stemmed prostheses. The main advantages of the humeral head resurfacing are the bone-preserving fixation and the relatively simple surgical technique. PMID- 11480091 TI - [Shoulder hemi-arthroplasty in proximal humeral fractures]. AB - Most fractures of the proximal humerus with significant displacement are best treated surgically. The range of surgical treatment varies from closed reduction and pinning to hemiarthroplasty depending on the degree of displacement, age of the patient, and bone quality. Determining whether or not the individual fractured bone segments are displaced to a significant degree requires high quality x-rays which can be difficult to obtain from acutely injured patients. Indications for replacement of the humeral head in acute fractures include: head splitting fractures in elderly patients, Neer 4-part fracture dislocations, selected 3 part fractures and fracture dislocations in elderly patients with poor bone quality and a very small head fragment, selected severe impression fractures in elderly patients that involve more than 40% to 50% of the articular surface and selected anatomical neck fractures in which internal fixation is not possible. If a prosthetic replacement of the humeral head is chosen, secure repair of the tuberosities is essential to avoid tuberosity migration and malunion. The clinical results of prosthetic replacement of the proximal humerus for acute fractures are superior to those for late arthroplasty. This treatment modality has been proven to relieve pain. However, even for patients treated with primary arthroplasty, a restricted range of motion has to be expected postoperatively. Furthermore, several studies indicate that a significant number of complications can occur following early and late prosthetic replacement. Humeral head replacement as a salvage procedure after malunions or failed open reduction and internal fixation is technically demanding with a relatively high rate of complications. Newer implant designs and instruments may improve the clinical results. PMID- 11480092 TI - [Treatment errors of metacarpal fractures. An analysis of expert testimony of the "Committee for Medical Malpractice" of the North Rhine Physician's Council]. AB - A large number of publications notwithstanding, there are no clear guidelines regarding the treatment of metacarpal fractures. Some authors believe that even severely dislocated fractures should not be surgically fixated. This paper analyzes the forensic problem on the basis of court rulings in Germany. Seventeen evaluations of metacarpal fractures conducted by the commission on medical malpractice of the physicians' council in North Rhein were analyzed and common mistakes were listed. The average age of the eight male and nine female patients was 41.9 years. The fifth metacarpal bone (MB) was affected eight times, the fourth MB four times, and the first MB twice. The second MB and third MB were affected once each. Surgery was performed in nine cases, while the conservative approach was taken eight times. An independent expert determined medical malpractice in 13 cases (76%). In 10 of these 13 cases, the symptoms the patients exhibited were linked to medical malpractice. In the 13 cases of medical malpractice there were 5 cases with technically insufficient osteosynthesis, 4 cases of uncorrected dislocation, 2 false diagnoses, and 2 false immobilizations. An independent expert identified a link to the symptoms in three of the four cases of uncorrected dislocation. These were an anatomically nonreduced fracture with joint involvement, a fracture with a dislocation of 50 degrees, and a fracture with rotation dislocation. Cases where fractures healed with severe dislocation or immobilization in a wrong position or for a too long a period are common but avoidable mistakes in the treatment of metacarpal fractures. PMID- 11480093 TI - Charleston, cocaine, and consent. PMID- 11480094 TI - Stroke risk in older men and women: aspirin, estrogen, exercise, vitamins, and other factors. AB - OBJECTIVE: To identify risk factors and preventive measures for stroke in elderly men and women. DESIGN: Observational prospective cohort study. SUBJECTS: Women (N = 8532) and men (N = 4722) between the ages of 44 and 101 (median age, 74) residing in a retirement community in southern California who had no previous history of stroke. METHODS: Upon entering the study in 1981, 1983, or 1985, study participants filled out a detailed health survey questionnaire about their medical history, exercise habits, intake of caffeinated beverages, alcohol, vitamins, and foods containing vitamins A or C, and history of smoking. Women also reported their use of estrogen replacement therapy. Cohort members were followed by periodic resurvey and by examination of death records through 1998. Age-adjusted stroke incidence rates, relative risks, and 2-sided P values were calculated. RESULTS: Between 1981 and 1998, 1211 women and 773 men were hospitalized for cerebrovascular disease. In women, risk of cerebral occlusion decreased significantly with increasing duration and recency of estrogen use. Hypertension, diabetes, heart attack, and smoking were significant stroke risk factors in both women and men. Exercise reduced stroke risk in general, and antioxidant vitamin supplements decreased the risk of cerebral occlusion. CONCLUSION: These results emphasize the role of lifestyle modification in the primary prevention of stroke and suggest that estrogen replacement therapy may be a potential preventive measure for women. PMID- 11480095 TI - Hirsutism: diagnosis and management. AB - The majority of women with hirsutism have the idiopathic variety characterized by normal circulating androgens. An index of suspicion should, however, be maintained for that subset of women whose hirsutism is indicative of a systemic hormonal aberration. The psychological consequences of this highly visible condition that can seriously impact quality of life substantiate the need for safer and more effective management options. The advent of a topical medication (eflornithine hydrochloride), devoid of systemic side effects, is one positive step toward this goal. PMID- 11480096 TI - Gender and osteoporosis: similarities and sex-specific differences. AB - Osteoporosis is a major health problem for men and women. Although women are disproportionately affected, the aging of the population and the exponential fracture increase in older persons ensure that the absolute numbers of osteoporotic fractures in both sexes will increase strikingly by the end of this decade. Important sex-specific differences are already known to occur in bone physiology and geometry, fracture epidemiology, bone gonadal hormone response, and post-hip-fracture mortality. These differences point to the importance of doing separate studies of osteoporosis in males and females and will hopefully lead to more effective and prevention-based strategies in the future. PMID- 11480097 TI - Testosterone deficiency and depression in aging men: pathogenic and therapeutic implications. PMID- 11480098 TI - Testosterone replacement in older men and women. AB - This article examines in detail the present state of the art concerning androgen deficiency in aging males. There is increasing evidence that testosterone replacement in hypogonadal older males can result in an improvement in quality of life. The major effects of testosterone are on libido, muscles, bone, and cognition. Less information is available concerning the role of testosterone in postmenopausal women, but testosterone replacement may have a role to play in treating disorders of libido and the sarcopenia that occurs at menopause. PMID- 11480099 TI - Treating symptoms of female sexual arousal disorder with the Eros-Clitoral Therapy Device. AB - OBJECTIVE: The aim of this study was to determine the effect of the Eros-Clitoral Therapy Device (Eros-CTD) on the sexual function of women with and without symptoms of female sexual arousal disorder (FSAD). DESIGN: Periodic survey of sexual function in women using the Eros-CTD over a six-week period. SUBJECTS: A total of 19 women participated in the study--10 with symptoms of FSAD and nine without symptoms of FSAD--ranging in age from 28 to 65 years, with a mean age of 45.2 years. METHODS: Ten patients with symptoms of FSAD and 10 without symptoms were instructed in the use of the small, portable vacuum device, Eros-CTD. One woman without symptoms of FSAD withdrew early in the study for personal reasons. The patients were instructed in the correct use of the device and were asked to complete one Female Intervention Efficacy Index (FIEI) each week. The patients also kept diaries of their use of the device, noting the frequency, length, and strength of vacuum. RESULTS: There was a significant improvement in all symptoms of FSAD (P < .05), including increased sensation, improved vaginal lubrication, enhanced ability to orgasm, and greater overall satisfaction. Patients without FSAD also reported similar changes in sensation, lubrication, ability to orgasm, and overall satisfaction. LIMITATIONS: This study was done on a small sample of self-selected patients, was of limited duration, and had no long-term follow-up. All of these factors should be considered in interpreting the data. CONCLUSION: The Eros-CTD was safe and effective in improving symptoms of FSAD in this group of women. Further studies on the efficacy of the Eros-CTD are indicated. PMID- 11480100 TI - Sex-related differences in hemostasis and thrombosis. AB - Disorders of thrombosis and hemostasis represent both diagnostic and therapeutic challenges for the clinician, in part because of sex-based differences in incidence and presentation. The hemophilias are characterized by specific sex linked patterns of inheritance, and there are sex differences in the presentation of the autosomally inherited disorders, particularly von Willebrand's disease. The diagnosis of these disorders can be affected by variations in either endogenous or exogenous estrogens, and the hemostatic stresses presented by menstruation and childbirth render any coagulopathy more severe in females than in males. Women are also at increased risk for developing thrombotic and embolic problems while on exogenous estrogens and during pregnancy. This article presents recommendations about the most appropriate and cost-effective ways to screen for the inherited disorders of both thrombosis and hemostasis in men and women. Recommendations are also developed for the treatment of women with these disorders, particularly in the context of pregnancy, contraception, uterine bleeding, and postmenopausal management. PMID- 11480101 TI - Hysterectomy and the female heart: understanding the risk. PMID- 11480102 TI - HRT may not reduce fracture in women without osteoporosis. PMID- 11480103 TI - Gender differences in depression associated with neurologic illness: clinical correlates and pharmacologic response. AB - Functional depression (i.e., depression without neuropathology) occurs approximately twice as often in women as in men. A review of the literature from the period 1966-1999 on the prevalence, clinical correlates, and treatment of depression in neurologic disease revealed a female preponderance of depression in diffuse neurologic disease, including Alzheimer's disease. In focal neurologic disease, the data were consistent for men and women, with a 1:1 ratio. Treatment data on depression in neurologic disease are scant, with the exception of poststroke depression. Although gender-based outcome data on the treatment of functional depression reveal better tolerability and response to serotonin reuptake inhibitors in women than in men, this phenomenon cannot be generalized to depression in neurologic disease. Men seem to consistently respond better than premenopausal women to tricyclic antidepressants in both functional and neurologic disease. Understanding how gender influences depression in neurologic illness and its response to treatment is a necessary step to improve the specificity of psychiatric treatment for depression. PMID- 11480104 TI - Advocates for "men's health": the newest voice. PMID- 11480105 TI - Lipid-lowering medications: what the new guidelines and data mean to women. PMID- 11480106 TI - [Chronic disease and therapeutic education]. PMID- 11480107 TI - [Exercise and inflammatory mediators]. PMID- 11480108 TI - [Peripheral muscles. Functional evaluation]. PMID- 11480109 TI - [Peripheral muscles: mechanical and metabolic function]. PMID- 11480110 TI - [Peripheral muscles and metabolic diseases]. PMID- 11480111 TI - [Peripheral muscles in chronic obstructive pulmonary disease]. PMID- 11480112 TI - [Peripheral muscles and cardiac insufficiency]. PMID- 11480113 TI - [Education and chronic respiratory disease]. PMID- 11480114 TI - [Peripheral muscles and corticosteroid therapy]. PMID- 11480115 TI - [Exercise retraining in severe chronic obstructive pulmonary disease]. PMID- 11480116 TI - [Exercise retraining in cardiac insufficiency]. PMID- 11480117 TI - [Exercise retraining in patients with type 2 diabetes: practical aspects]. PMID- 11480118 TI - [Physical activity and aging]. PMID- 11480119 TI - [Physical activity and public health]. PMID- 11480120 TI - [Education and respiratory rehabilitation]. PMID- 11480121 TI - [Mechanisms of dyspnea in chronic cardiac insufficiency. Dyspnea in cardiac insufficiency]. PMID- 11480122 TI - Infectious mononucleosis: not always what it seems. PMID- 11480123 TI - The challenge of multidrug resistant tuberculosis. PMID- 11480124 TI - Benign salivary gland disease. AB - The majority of benign clinical problems that present affect the major salivary glands--parotid and submandibular. However, there are numerous minor salivary glands located in the mucosa of the head and neck which have the same predilection to the same diseases that affect the major glands but to a lesser frequency. PMID- 11480125 TI - Salivary gland stones: diagnosis and treatment. AB - Salivary calculi are a common cause of salivary gland disorder and may occur in any of the salivary glands and at almost any age. The stones may be small and intraductal or lie within the gland substance when they may become very large. They cause symptoms by obstructing salivary flow. Diagnosis is usually straightforward, and treatment is aimed at stone removal. PMID- 11480126 TI - Management of malignant salivary gland tumours. AB - Malignant tumours of both major and minor salivary glands are extremely rare: the parotid gland is most frequently affected by malignant change, followed by the submandibular gland. Malignant tumours of minor salivary glands may occur anywhere in the upper aerodigestive tract, but more common sites are the upper lip, junction of hard and soft palate and paranasal sinuses. PMID- 11480127 TI - Pneumococcal disease in childhood. AB - Streptococcus pneumoniae is the most common cause of serious bacterial infections in children worldwide. Problems with antibiotic resistance have lead to changes in antibiotic policies for children with possible pneumococcal disease. Demonstration of the efficacy of a pneumococcal conjugate vaccine has lead to consideration of its inclusion in routine infant vaccination schedules. PMID- 11480128 TI - Electrocardiogram interpretation. Case 4: Collapse as a result of bradycardia. PMID- 11480129 TI - Actos (pioglitazone): a new treatment for type 2 diabetes. AB - Type 2 diabetes is increasingly common and can be difficult to control. By directly targeting insulin resistance, the thiazolidinediones offer a new mode of treatment. Here, the pharmacology, clinical trial evidence, side-effects and current clinical uses of pioglitazone are reviewed. PMID- 11480130 TI - Nasal manifestations of granulomatous disease. AB - Granulomatous disease frequently affects the head and neck region, particularly the nose and sinuses. This article describes the most common infectious and non infectious conditions and their clinical features. PMID- 11480131 TI - Treatment of acute porphyria. AB - Haem preparations are now available for the specific treatment of attacks of acute porphyria. This review focuses on their use in this uncommon but life threatening medical emergency. PMID- 11480132 TI - How to write a critical letter and respond to one. AB - Critical letters are important. They correct the published record. To write a critical letter requires tact; to respond to one requires tact--and the humility that comes with the realisation that belief in an observation or an idea does not make it true. PMID- 11480133 TI - Skills training using multimedia and models. AB - All doctors need basic surgical skills. Learning and retaining such skills requires repeated practice. This article describes structured training provided by a multimedia computer program combined with practice on simulated tissue models. PMID- 11480134 TI - Pseudomembranous colitis without diarrhoea following Helicobacter pylori eradication therapy. PMID- 11480135 TI - Haemorrhagic rash in infectious mononucleosis. PMID- 11480136 TI - A case of spontaneous cervical extradural haematoma. PMID- 11480137 TI - The lost tribes: finding our way out of the darkness. PMID- 11480138 TI - Why use non-invasive ventilation in acute respiratory failure? PMID- 11480139 TI - Scars: the battle over isotretinoin. PMID- 11480140 TI - Aquatic antagonists: lionfish stings. AB - Although lionfish can be found in all the oceans, the highest incidences of human stings appear to be in the tropics, especially in the Indo-Pacific area and Mediterranean Sea. The recent interest in tropical fish aquaria has expanded the geographic range of the stings of these animals. PMID- 11480141 TI - Childhood herpes zoster. AB - Herpes zoster (HZ) in childhood is rather unusual. This reactivation of chickenpox, the primary varicella-zoster virus (VZV) infection that lies dormant within sensory ganglia, is seen with increased frequency in otherwise healthy children who acquire chickenpox either in utero or within the first year of life. Our patient is a good example of this; he was exposed to chickenpox at the age of 3 months (by his 2 siblings) and developed HZ at 6 years of age. PMID- 11480142 TI - Photo quiz. What is your diagnosis? Fordyce spots. PMID- 11480143 TI - Erythema dyschromicum perstans: a case report and review. AB - Erythema dyschromicum perstans (EDP) is an acquired ashy dermatosis characterized by patches of blue-gray pigmentation over the trunk, extremities, and neck. These pigmentary changes may become chronic and disfiguring. At present, the etiology of EDP remains unclear, and there is no single established therapy. We describe a 21-year-old Hispanic man with extensive patches of EDP who improved with oral corticosteroid therapy. PMID- 11480144 TI - Pellagra: a sporadic pediatric case with a full triad of symptoms. AB - Pellagra is clinically manifested by a photosensitive dermatitis, diarrhea, and dementia. The full triad of symptoms is usually not well developed in infants and children. We report a case of a 14-year-old boy with classic symptoms of pellagra. All his symptoms responded to treatment with nicotinic acid. PMID- 11480145 TI - Management of male pattern hair loss. AB - The management of androgenetic alopecia (AGA) has been materially altered by the availability of the 5 alpha-reductase type 2 inhibitor, finasteride. Nevertheless, this agent is only one component of successful management, and an understanding of the role of camouflage agents, surgical options, and other medical treatments is important. Because no treatment completely reverses baldness, it is important to communicate the limitations of each modality to the patient so that he has appropriate expectations of the outcome of any intervention. Patient counseling and support are also often relevant. PMID- 11480146 TI - Erythema elevatum diutinum in a patient with human immunodeficiency virus. AB - Erythema elevatum diutinum (EED) is a chronic cutaneous leukocytoclastic vasculitis. This rare disease is characterized by red, brownish-purple, and yellow papules, plaques, and nodules distributed symmetrically about the extremities. There have been recent reports of the disease in association with infection with the human immunodeficiency virus (HIV). We describe the case of a 51-year-old man with HIV who presented with EED, which was successfully treated with dapsone. PMID- 11480147 TI - Vinegar as a disarming agent to prevent further discharge of the nematocysts of the stinging hydromedusa Olindias sambaquiensis. AB - Olindias sambaquiensis, known as agua viva, is an endemic hydromedusa that is distributed from latitude 23 degrees to 42 degrees south in the southwestern Atlantic Ocean and is known to cause severe stings. This is the first report of the use of vinegar in disarming O sambaquiensis nematocysts and includes preliminary observations on its use in managing the stings of hydromedusae. PMID- 11480148 TI - Classic Kaposi sarcoma presenting as rapidly growing nodules. AB - Classic Kaposi sarcoma (KS) is a sporadic disease that usually affects persons older than 50 years, with a distinct male predominance. Although classic KS has a protracted, indolent course, there appears to be a rare disseminated fulminant type. This report describes a case of classic KS, presenting as a very rapid enlargement and dissemination of skin lesions, without evidence of human immunodeficiency virus (HIV) infection or involvement of other internal organs. PMID- 11480149 TI - Complications of eyelash and eyebrow tattooing: reports of 2 cases of pigment fanning. AB - Eyelash and eyebrow tattooing are commonly performed procedures that have a very low rate of reported complications. We describe one case of infraorbital pigmentation after eyelash tattooing and another of periorbital pigmentation after eyebrow tattooing. Although most complications related to eyelash and eyebrow tattooing, including pigment fanning, have been reported by ophthalmologists, pigment fanning is also of concern to dermatologists. PMID- 11480150 TI - Solitary nodule of the great toe. AB - We describe a 21-year-old woman with a subungual exostosis exhibiting both skin and nail findings. The patient presented with a firm, flesh-colored, nontender, subungual nodule in the distal nail bed of the great toe. Radiographic examination revealed focal calcification of the nodule, with direct communication to the underlying phalanx. Subungual exostosis should be considered in the differential diagnosis of any digital mass. Surgical excision, followed by curettage of the base, is the treatment of choice. PMID- 11480151 TI - Prednicarbate emollient cream 0.1% in pediatric patients with atopic dermatitis. AB - Atopic dermatitis, common among infants and children, is an intensely pruritic, chronic, inflammatory dermatosis that is traditionally treated with emollients for dry skin and topical corticosteroids for inflamed areas. A multicenter, 3 week, open-label study evaluated prednicarbate emollient cream 0.1%, a nonhalogenated midpotency corticosteroid, in 55 patients aged 4 months to 12 years who were diagnosed with atopic dermatitis. No suppression of the hypothalamic-pituitary-adrenal (HPA) axis was evidenced by serum cortisol levels obtained before and after intravenous injection of 250 mg of cosyntropin on days 1 and 22, and biochemical tests detected no other systemic effects. Adverse events were few and within the expected range. Prednicarbate resulted in improvements based on global evaluations and sign/symptom scores. In conclusion, this study found prednicarbate emollient cream 0.1% to be safe and effective for the treatment of atopic dermatitis in pediatric patients for up to 3 weeks. PMID- 11480152 TI - [Implanting more defibrillators. How and where?]. PMID- 11480153 TI - [Aortic atheroma: not a sensitive indicators for coronary artery disease in patients with mitral valve disease]. AB - Aortic atheroma detected by transoesophageal echocardiography has been reported to be a good prognostic marker for coronary disease on angiography. The value of this detection in valvular heart disease would be to avoid preoperative coronary angiography in asymptomatic patients. The aim of this study was to assess the prognostic value of aortic atheroma in a population with a low prevalence of coronary artery disease in whom transoesophageal echocardiography was systematically performed. In addition, calcification of the aortic knuckle, a marker of atherosclerosis, was analysed by simple chest X-ray. One hundred and ninety two patients (103 men, 89 women; mean age: 63.1 +/- 15 years), operated for mitral valve replacement, underwent transoesophageal echocardiography, angiography, within 6 months, and chest X-ray. The cardiovascular risk factors, presence of aortic atherome, angiographic coronary artery disease and aortic calcification were studied. Aortic atheroma was observed in 72 patients (37.5%), usually in the descending thoracic aorta (73.6%). Coronary stenosis was observed in 36 patients (18.7%). On univariate analysis, aortic atherome predicted coronary stenosis with a sensitivity of 53%, specificity of 66% and positive predictive value of 26% and negative predictive value of 86%, compared with chest X-ray: 71%, 65%, 33% and 90%, respectively. In multivariate analysis, only hypercholesterolaemia, smoking and age predicted the presence of coronary artery disease. The presence of aortic atheroma was not predictive (p = 0.3). The authors conclude that aortic atheroma does not predict the presence of coronary artery disease in a patient population with mitral valve disease and a low prevalence of coronary artery disease. Simple chest X-ray has almost the same diagnostic value. The association of these two investigations does not give sufficient negative predictive values to avoid coronary angiography. PMID- 11480154 TI - [Coronary surgery of the beating heart. First year of experience in a series of 317 non-selected coronary patients]. AB - From May 1999 to May 2000, 317 unselected patients, representing 92.7% of all coronary artery surgery procedures, underwent open heart surgery of the beating heart by median sternotomy with the aid of a cardiac stabilising device. The main preoperative characteristics were: mean age = 66.1 years; men = 78.9%; left main stem disease = 31.8%; mean left ventricular ejection fraction = 54.1%; mean Parsonnet index = 16.9. These 317 patients were compared with a group of 303 patients who underwent coronary bypass surgery the year before by the same surgical team with cardiopulmonary bypass (CPB) and cardiac standstill. Seven hundred and eighty-six distal anastomoses were carried out in the beating heart group (2.48 grafts per patient) compared with 2.91 in the CPB group: p < 0.001). There were 10.1% single bypass, 37.5% double bypass, 47.3% triple bypass and 5% quadruple bypass procedures. A cardiopulmonary bypass was required in 13 patients (4.1%). The mortality at 30 days was 3.1% versus 4.6% in the CPB group (p = NS). The need for blood transfusion was reduced by nearly 40% in the beating heart group (23.7% versus 39.9%, p < 0.001). The incidence of cerebrovascular complications was reduced from 3% in the CPB group to 0.6% in the beating heart group (p = 0.06). The peak postoperative troponine I levels were much lower in the beating heart group (2.5 versus 6.4 ng/ml, p < 0.001). The authors conclude that surgery on the beating heart is feasible in most patients. Compared with conventional surgery under CPB, there seems to be less requirement for blood transfusion and a tendency to reduce the cerebral risk. Nevertheless, a large prospective randomised trial is required to validate the potential advantages and limitations of this technique with respect to conventional surgery and to determine the optimal indications of surgery on the beating heart. PMID- 11480155 TI - [Videosurgical pleuro-pericardial windows in oncology]. AB - Fifteen consecutive patients with recurrent pericardial effusion associated with confirmed neoplastic disease (N = 11) or with a triad of symptoms: weight loss, anorexia, tiredness (N = 4), underwent videosurgery through a pleuro-pericardial window. The mean age was 58 years (37-77 years). The average procedure and assisted ventilation times were 56.5 min (40-110 min) and 86 min (70-140 min) respectively. Three patients experienced cardiac arrhythmias which regressed. The patients were discharged home on the 5th day (3-11 days). In contrast to needle biopsy which only provided a diagnosis in 3 cases, the histopathological findings were diagnostic in all cases: 12 malignant and 3 benign pericardial effusions with correction of the presumed clinical diagnosis in the latter 3 cases. The average follow-up was 10 months (7 days-3.5 years). There was no operative mortality. Global survival at 1 year was 66%, death being generally caused by a complication of the malignant disease. There were no deaths in the 3 patients with benign pericardial effusions, underlying the necessity of an accurate etiologic diagnosis before assuming malignant pericardial invasion. There were no recurrences of the pericardial effusion and the ventilatory status of these patients was adapted for everyday activities. In patients with malignant disease, the construction of a pleuro-pericardial window by videosurgery is a satisfactory approach. It provides etiologic diagnosis and is well tolerated by patients in poor general condition with no operative deaths, low morbidity and definite improvement in the patients' comfort. PMID- 11480156 TI - [Identification of unstable coronary atherosclerotic plaques]. AB - The mechanisms of atherogenesis are better understood and the detection of atherosclerosis has improved with the different diagnostic methods currently available. However, it is almost impossible at present to differentiate high risk, unstable or vulnerable plaques from quiescent or stable plaques of atherosclerosis. This is a crucial problem given the banality of atherosclerosis on the one hand, and, on the other hand, the serious consequences (acute coronary syndromes, cerebrovascular accidents) of thrombotic occlusion at the site of an atherosclerotic plaque. It has now been established that the composition of the plaque is more important than the degree of stenosis, a fundamental concept in the risk of plaque rupture, precipitating the cascade of reactions leading to uncontrolled thrombosis. Consequently, new imaging techniques should address the problem of analysing the composition of atheromatous plaques. Endovascular ultrasonography, fast CT, angioscopy, nuclear imaging techniques and MRI are so many promising tools. However, non-invasive techniques should be distinguished from invasive ones. In all probability, it will be the former which will turn out to be the most useful diagnostic aid in pauci or asymptomatic patients. This article reviews the different imaging techniques under evaluation for the identification of risk of plaque rupture. PMID- 11480157 TI - [Mediators of reactive hyperemia]. AB - Reactive hyperemia is a protective adaptive phenomenon, which quickly restores blood flow distal to a transient arterial occlusion. It involves the vascular tone regulation mechanisms of the ischaemic distal territory as a whole, but also its proximal control (flow-mediated dilation). The resulting vasodilatation differs from one vascular bed to another, and also between the large proximal arteries and the distal arterioles in the microcirculation. Thus, although both are elicited by a similar transient occlusion of brachial blood flow, non invasive investigation of reactive hyperemia in the humeral circulation differs from assessment of endothelial function with the flow-mediated arterial dilation. Indeed, whereas nitric oxide (NO) plays the key-role in the later, several mediators are involved in the former, including myogenic and metabolic factors, prostaglandins, K + ATP channels, adenosine and NO which is only one of the many components of this complex regulation mechanism. PMID- 11480158 TI - [The heart and cocaine]. AB - Cocaine abuse should be considered in everyday cardiological practice for three reasons. The first one is epidemiological as there is an unprecedented increase in cocaine abuse in France and it may result in serious complications, mainly cardiovascular. The second reason concerns modality of consumption: it is usually associated with other toxic substances (including tobacco) and poly-intoxication is a more serious problem and the diagnostic difficulties are greater. Finally, chest pain is the first symptom of myocardial infarction in about 6% of cases. In cardiological practice, young adults with this symptom should be suspected of cocaine abuse and the diagnosis should be eventually confirmed by toxicological analysis. PMID- 11480159 TI - [Mechanical and functional predictive factors for restenosis and arterial remodeling after experimental angioplasty]. AB - Arterial remodelling plays an important part in post-angioplasty restenosis but the physiopathology of this process is not fully understood. Abundant collagen synthesis and endothelial dysfunction have been demonstrated after angioplasty, but their role in restenosis and remodelling has not been studied. The aim of this study was therefore to assess endothelial function and collagen with respect to the severity of restenosis and the type of arterial remodelling. Atherosclerosis was induced by an association of endothelial abrasion and a high cholesterol diet in the femoral arteries of 22 white New Zealand rabbits. Four weeks later, angioplasty was performed. The acetylcholine endothelium-dependant vasomotricity (expressed as % inhibition of contraction to phenylephrine), collagen and morphology were assessed 28 days after angioplasty. The change in acetylcholine endothelium-dependant vasomotricity was greater in severe restenosis (r = 0.61, p = 0.02). Endothelium-dependant relaxation was not significantly altered when remodelling was expansive and very abnormal when it was constrictive (35.5 +/- 13.0 vs 3.7 +/- 7.9%; p = 0.04). Restenosis was associated with an increase in collagen (r = 0.69, p = 0.004). The density of collagen was significantly higher in constrictive remodelling than in expansive remodelling (34.5 +/- 4.5 vs 18.2 +/- 4.7%; p = 0.03). Endothelial dysfunction and collagen accumulation are correlated with the severity of restenosis and with constrictive remodelling after angioplasty in an experimental model. PMID- 11480160 TI - [Atrial flutter: a possible early sign of acute rejection in heart transplantation. A case report]. AB - The authors report the case of a cardiac transplant patient with a recurrence of atrial flutter two months after electrical cardioversion and despite long-term preventive treatment with amiodarone. Early investigation for signs of rejection with 4 endomyocardial biopsies was negative. Aggravation of the haemodynamic status due to flutter with a rapid ventricular response led to an attempted radio frequency ablation. Endocavitary mapping confirmed persistence of sinus activity in the native atrium and the presence of a circuit of type I isthmic flutter (anticlockwise circuit) in the donor atrium. Ablation by radio-frequency in the same procedure was successful. A fifth myocardial biopsy the same day finally confirmed stage 3A acute rejection. No signs of recurrent rejection or arrhythmia have been observed after 24 months' follow-up in this patient. This preliminary experience confirms the need to look for graft rejection by repeated myocardial biopsies in cardiac transplant, patients with atrial flutter and the efficacy of radio-frequency ablation in cases of resistance to conventional therapy. PMID- 11480161 TI - [Recurrent rhythmic cardiomyopathy secondary to idiopathic ventricular tachycardia]. AB - Tachycardia-induced cardiomyopathy consists in severe ventricular dysfunction and heart failure secondary to incessant or prolonged supraventricular or less frequently ventricular tachycardia in the absence of any cardiac disease. We present here a case of recurrent severe cardiomyopathy secondary to idiopathic ventricular tachycardia. Complete sustained recovery in function was observed after successful radiofrequency ablation of the ventricular tachycardia focus. To the best of our knowledge, this is the first report of recurrent tachycardia induced cardiomyopathy. The diagnosis, aetiology, physiopathology and mechanisms of tachycardia-induced cardiomyopathy are discussed. Several therapeutic implications are presented. PMID- 11480162 TI - Interactive effects of the affect quality and directional focus of mental imagery on pain analgesia. AB - College students (25 men and 25 women) were randomly assigned (within sex) to each of the 4 factorial groups, based on manipulation of affect quality (positive vs. negative) and directional focus (internal vs. external) of mental imagery, and to a control group receiving no manipulation. Both imagery variables had a significant impact on pain tolerance and ratings during a cold-pressor test with positive affect and external imagery producing greater analgesia than their counterpart conditions. Positive affect imagery combined with external imagery resulted in the lowest reported pain amongst the groups. However, self-reported mood descriptors did not consistently parallel the pain tolerance and rating data. Likewise, although heart rate and skin potential responses increased during the cold pressor for the group as a whole, the only significant difference amongst the experimental groups was the relatively higher skin potential reactivity of the positive affect-external imagery group--possibly reflecting greater task engagement for this group. Seemingly, imagery in this situation operates primarily via cognitive, rather than via physiological mediators of the pain experience. PMID- 11480163 TI - A psychophysiological marker of attention deficit/hyperactivity disorder (ADHD)- defining the EEG consistency index. AB - This study continues our research to further validate the idea that ADHD (Attention Deficit/Hyperactivity Disorder) interferes with transition from one task to another and this interference can be quantified by a Consistency Index (CI) derived from a specific mathematical representation of EEG data. We reanalyze 32 previously reported data sets present new data for 35 boys and girls, ages 7-12, ADHD or control. Each data set contains EEG, recorded and digitized while participants perform consecutive 10-min tasks: video, reading, and math. For boys, the CI in ADHD was four times lower than in controls, p < .005, for girls this difference was two times, p < .05. ADHD/control classification based on the CI coincided with the DSM-IV criteria for 88% of the boys and for 67% of the girls. Post hoc analysis indicated that the classification utility of the CI diminished with age. A CI below 40% could be a discriminating, reliable, and reproducible marker of ADHD in young boys. PMID- 11480164 TI - Treatment of phantom limb pain with combined EMG and thermal biofeedback: a case report. AB - Phantom pain is a frequent consequence of the amputation of an extremity and causes considerable discomfort and disruption of daily activities. This study describes a patient with extreme phantom limb pain following amputation of the right upper limb. The treatment consisted of 6 sessions of EMG biofeedback followed by 6 sessions of temperature biofeedback. The patient did not use a prosthesis and had not received previous treatment for chronic pain. Results demonstrated complete elimination of phantom limb pain after treatment, which was maintained at a 3- and 12-month follow-up. Pain relief covaried with increase in skin temperature at stump and perceptual telescoping (retraction of phantom limb into stump). PMID- 11480165 TI - The physiological correlates of Kundalini Yoga meditation: a study of a yoga master. AB - This study explores the physiological correlates of a highly practiced Kundalini Yoga meditator. Thoracic and abdominal breathing patterns, heart rate (HR), occipital parietal electroencephalograph (EEG), skin conductance level (SCL), and blood volume pulse (BVP) were monitored during prebaseline, meditation, and postbaseline periods. Visual analyses of the data showed a decrease in respiration rate during the meditation from a mean of 11 breaths/min for the pre- and 13 breaths/min for the postbaseline to a mean of 5 breaths/min during the meditation, with a predominance of abdominal/diaphragmatic breathing. There was also more alpha EEG activity during the meditation (M = 1.71 microV) compared to the pre- (M = .47 microV) and postbaseline (M = .78 microV) periods, and an increase in theta EEG activity immediately following the meditation (M = .62 microV) compared to the pre-baseline and meditative periods (each with M = .26 microV). These findings suggest that a shift in breathing patterns may contribute to the development of alpha EEG, and those patterns need to be investigated further. PMID- 11480166 TI - From tibialis anterior to Tai Chi: biofeedback and beyond. AB - This keynote presentation highlights events that have contributed to scientific explorations of one research clinician. Steve Wolf traces his scientific roots to early studies in single motor unit control under the guidance of his primary mentor, John Basmajian, MD. This work led to subsequent studies on the role of EMG feedback in predicting successful outcomes in upper extremity use and in ambulatory capabilities among patients with chronic stroke. These findings are contrasted to further efforts to condition entire reflexes rather than individual muscles through use of operant-conditioning paradigms. The findings from applications of EMG biofeedback to stroke patients became the basis for minimal motor criteria in the treatment of the impaired upper extremities of patients with chronic stroke, using "forced use" or "constraint-induced movement therapy." Last, investigations into center of pressure feedback using computerized balance machines resulted in a series of experiments that ultimately led to the finding that Tai Chi as an exercise form for older adults can have a substantially favorable effect in delaying the onset of fall events. PMID- 11480167 TI - Psychophysiological reactivity of aggressive drivers: an exploratory study. AB - In the United States, motor vehicle accidents are the leading cause of accidental death and injury. Aggressive driving, which has been identified as a major risk factor for motor vehicle accidents by transportation authorities, is thus an important topic of study. This study compared the physiological reactivity of self-referred aggressive and nonaggressive drivers. Heart rate, blood pressure, facial muscle activity, and skin resistance were monitored as participants listened to idiosyncratic vignettes of driving and fear-provoking scenarios, as well as during a standard stressor task (mental arithmetic). The results were that aggressive drivers exhibited significant increases in muscle tension and blood pressure during the driving vignettes, relative to controls. They also responded to the fear vignette and mental arithmetic in a qualitatively different fashion from that of controls. The aggressive drivers responded to these stimuli with less overall heart rate and electrodermal reactivity, but increased blood pressure and muscle tension. In contrast, the controls responded to the fear vignette and mental arithmetic primarily with increased heart rates and decreased skin resistance. The findings suggest that both physiological hyperarousal as well as differential responses to stressful stimuli may contribute to aggressive driving. Implications for interventions with this population are discussed. PMID- 11480168 TI - [Detection of numerical chromosome aberrations in sperm of workers exposed to benzene series by two-color fluorescence in situ hybridization]. AB - To study the frequency of numerical chromosome aberrations in sperm of workers exposed to benzene series, the aneuploidy frequencies of 9 and 18 chromosomes were detected in sperm of workers by two color fluorescence in situ hybridization with digoxingenin labeled 9 chromosome probe (D9z1) and biotin labelled 18 chromosome probe (D18z1). The timeweight average air concentration (TWA) of benzene in the workplace was 86.49 mg/m3, it was two fold higher than the national maximum allowable concentration. The concentration of urinary trans, transmuconic acid (ttMA) in exposed group was significantly higher than that of control group. The sperms counted in 14 exposed workers and 16 control workers were 136,401 and 156,955 respectively. The results showed that the disomy frequencies of sperm for chromosome 9 and 18 (0.168% +/- 0.063% and 0.055% +/- 0.031% respectively) and the frequency of diploidy sperm (0.073% +/- 0.045%) were statistically increased over that of the control group (0.050% +/- 0.030%; 0.033% +/- 0.025% and 0.040% +/- 0.036%, respectively). The nullisomic frequencies of 9 and 18 chromosome in the exposed group (0.206% +/- 0.047%, 0.068% +/- 0.044%) were statistically increased over that of the control group too (0.067% +/- 0.037%, 0.048% +/- 0.034%). The frequency of overall numerical chromosome aberrations in the exposed group was 0.570% +/- 0.144%, when this value was compared with that of control (0.218% +/- 0.071%), a statistically significance was presented. Our experiments showed that exposed to benzene at higher concentration may induce increase in aneuploidy frequency of sperm autosomal chromosome in exposed workers. PMID- 11480169 TI - [Analysis of association between gene polymorphisms of microsomal epoxide hydrolase (EPHX1) and infant birthweight]. AB - We investigated association between genetic polymorphisms of EPHX1 in mother and infant birthweight. Data of 342 female workers were collected in textile mill. A total of 342 mothers were genotyped for the His139Arg polymorphism of EPHX1 by a polymerase chain reaction-restriction fragment length polymorphism assay. Using multiple linear regression models, we estimated the adjusted association between polymorphisms of EPHX1 and birthweight, with adjusted for potential confounders. We found that polymorphisms of EPHX1 were closely associated with reduced birthweight (beta +/- SE = -149 g +/- 56, P = 0.0083) after adjustment for major cofounders. In subgroup of passive smoking analysis, the polymorphisms of EPHX1 were highly associted with birthweight for those with passive smoking (beta +/- SE = -234 g +/- 88, P = 0.0088); The significant association of EPHX1 polymorphisms with reduced birthweight were showed for those with working stress (beta +/- SE = -157 g +/- 59, P = 0.0079). Our data provide polymorphisms of EPHX1 in mothers were significant association with birthweight of their infant, and showed gene-environmental interaction in relation to birthweight. PMID- 11480170 TI - [Synergistic effect of the expression of p16INK4 gene and retinoblastoma gene in lung cancer]. AB - In order to clarify the frequency of p16INK4 gene inactivation and its relationship with retinoblastoma (Rb) gene expression in non-small cell lung cancer, in situ double hybridization was established with Bio-labelled Rb cDNA probe and Dig-lablled p16INK4 cDNA probe, and immunohistochemistry was carried out with polyclone rabbit antibodies to human p16INK4 and Rb protein. The results showed that positive hybridization of p16INK4 mRNA showed blue with the detecting system of alkiline phosphatase NBT/BCIP, positive hybridization of Rb mRNA showed red with the detecting system of HPR-AEC. The negative ratios of p16INK4 and Rb gene transcription were 22.6%(7/31) and 16.1%(5/31). The results of immunohistochemistry showed that the negative ratio of p16INK4 protein was 42%(13/31) with p16INK4 polyclone antibody, the negative ratio of pRb was 19.4%(6/31) with polyclone antibody. The data indicated that p16INK4 gene and Rb gene are synergistic in the progression of lung cancer, p16INK4 gene plays a significant role in the development of non-small cell lung cancer. PMID- 11480171 TI - [Studies of genetic diversity of Zhejiang native chicken breeds]. AB - A total of 539 bases of mitochondrial DNA D-loop region of five native chicken breeds of Zhejiang province and the white leghorn chicken (as the control) were sequenced by DNA sequencing technique, and the phylogenetic trees of the chicken breeds were constructed. The results showed that 24 variation sites i.e. 4.45% sequence divergence were detected among the 30 DNA sequences, and the six breeds belonged to two different maternal lineage, one included Xianju chicken and White Leghorn chicken which had the same maternal origin, the other included Lingkun chicken, Baiyiner chicken, Wugu chicken and Xiaoshan chicken which had the same matriarchic ancestor. Among the latter lineage, Lingkun chicken, Baiyiner chicken and Wugu chicken had a closer relationship to each other than to Xiaoshan chicken. PMID- 11480172 TI - [Cloning and sequencing of prolactin gene cDNA in three chicken breeds]. AB - The total RNA was extracted from the pituitary of two Chinese native chicken breeds, Yuehuang and Taihe Silkies, and one layer Isa, using RNeasy Kit (QIAGEN, Germany). The total RNA was used to synthesize a specific fragment with RT-PCR, in which the primers were designed based on the sequence of broiler prolactin gene cDNA. The specific fragment was ligated to a linear plasmid pBSSK and cloned with XL1-Blue. The sequencing of prolactin cDNA was carried out with ABI PRISH 377XL DNA Sequencer after cloning. The cDNA sequences and deduced amino acid sequences of prolactin gene of two Chinese native chicken breeds and one layer were compared with that of broiler, dwarf chicken and turkey. There was 93.97% 99.89% cDNA sequence homology among Chinese native, layer, broiler and dwarf chickens, in which there was the highest (99.87%) between those of Taihe and dwarf chickens. There was 98.25%-100% deduced amino acid sequence homology among Chinese native, layer, broiler and dwarf chickens, in which there was the highest (100%) between those of Taihe Silkies and dwarf chickens. It was found that Yuehuang and Taihe Silkies had the same signal peptide cleavage site Leu-Pro-Ile Cys among amino acids sequence deduced from pre-prolactin cDNA as broiler, dwarf and turkey, while layer Isa had a different cleavage site Pro-Pro-Ile-Cys. Such difference might cause a different translation processing of pre-prolactin, which could make layer Isa non-broody. There were different amino acids in the positions 71, 141, 150 and 175 of deduced prolactin amino acid sequences among Yuehuang, Taihe Silkies, layer, broiler and dwarf chickens. There was a heparin binding site in positions 175-181 (L-R-R-D-S-H-K) among prolactin amino acid sequences of broiler and Taihe Silkies. PMID- 11480173 TI - [Studies of the correlation between fragile sites and giant panda's breeding problem]. AB - An unique culture system has been established which can effectively indicate fragile sites induced by BrdU on giant panda chromosomes. After we cultured the lymphocytes of giant panda for 96 hours and treated it with low concentration of BrdU (10 micrograms/ml) for only a short time (4 hours), the areas of high frequency-expressed fragile sites were successfully located on the centromeres of No. 2 and No. 12 chromosomes. After statistic analysis, we found that the frequency of fragile sites on No. 2 and No. 12 chromosomes are notably different among pandas. What is more important, the former is relative to giant pandas' breeding conditions and correlation coefficiency (r equals to -0.772). The result indicates that high frequency of fragile sites on No. 2 chromosome is unfavorable to giant panda's breeding and survival of filial generation. PMID- 11480174 TI - [Genetic analysis of dominant earliness of rice genic male sterile line 6442S-7]. AB - F1 progenies of a genic male sterile and early maturing indica line 6442S-7 crossed with sixteen medium or late maturing varieties, such as Minghui 63, shared the same heading date as 6442S-7 under long day and short day, suggesting that 6442S-7 has completely dominant earliness. Segregation of heading dates of the F2 and B1F1 populations indicated that the earliness of 6442S-7 is mainly controlled by two dominant major genes. Whereas, heading dates of F1 progenies of 6442S-7 crossed with other four late maturing varieties, i.e. IR68, Xianguo, 9311, and BG1639, closed to the mid-parent value under long day. In F2 populations of 6442S-7 crossed with each of the four late maturing varieties, F1 from three-way crosses between 6442S-7 and each F1 of Minghui 63 crossed with each of the four late maturing varieties, and F1 from three-way crosses between 6442S-7 and each F1 of crosses between each two of the four late maturing varieties, segregation of heading dates showed that an incompletely diminant allelic suppressor for the dominant earliness of 6442S-7 existed in the late maturing varieties, IR68, Xianguo, 9311, and BG1639. It is considered that dominant earliness genes harbored in 6442S-7 have splendid future in rice improvement. PMID- 11480175 TI - [Evolution study of wheat (Tritium aestivum L.) A, B and D genome based on DNA sequence similarity]. AB - Tritium aestivum L., and its four potential origin species T. urartu T., T. monococcum L., Ae. squarrosa L. and T. dicoccum S., were amplified with MWG913, a STS marker located in 1H chromosome of barley. Those products are cloned and sequenced. By the discrepancies of these sequences, we have obtained some evidences about the origin of wheat A, B, D genomes at molecular level. The results showed that the studied sequence in A genomes of Tritium aestivum L. was identical with that of T. urartu T., T. monococcum L. and T. dicoccum S.; that in D genome of Tritium aestivum L. was as same as that of Ae. squarrosa L.; that in B genome of Tritium aestivum L. differed from that of T. dicoccum S. in 0.61%. The results also suggested that A, B and D genome have the different evolution speed. PMID- 11480176 TI - [Pyramiding and marker-assisted selection for powdery mildew resistance genes in common wheat]. AB - This project used the strategy of discarding susceptible individual plants and keeping resistance ones by resistance identification in breeding populations and planted resistant plants next in earlier generations (F1-F3), and performing marker-assisted selection combining resistance identification in later generation (F4 generation) for pyramiding powdery mildew resistance. F4 populations from crosses of elite parents possessing different Pm genes were screened by using molecular markers tightly linked to Pm2, Pm4a, Pm8 and Pm21 genes, respectively. Fourteen individual plants with Pm4a and Pm21 out of 40 F4 plants were identified. Sixteen individual plants containing Pm2 and Pm4a out of 80 F4 plants and six plants possessing Pm8 + Pm21 out of 58 F2 plants were screened. Notably, the plants with Pm2 and Pm4a showed high level resistance or immunity to powdery mildew, while plants with single Pm2 or Pm4a had lower resistance. The results suggest that the strategy of pyramiding different Pm genes provide an alternative way of utilizing resistance gene resources for breeding new types of resistance lines and cultivars, which will have significance not only in breeding practice but also in theoretical research. PMID- 11480177 TI - [The cytogenetical studies of fertility restoration of some male sterile lines of alloplasmic wheat]. AB - The frequency of cells with univalent (FCU) at MI and with lagging chromosomes and chromosomal bridge (FCLB) at AI in PMC of F1 hybrids of 4 alloplasmic 1BL:/1RS and non-1BL/1RS wheat male sterile lines with Aegilops kotschyi, Ae. variabilis, Ae. ventricosa and Ae. bicornis cytoplasms were systematically investigated, and the relationship between FCU at MI, FCLB at AI and F1 selfed seed set was analyzed. The results were as follows: (1) The frequency of abnormal chromosomes at MI and AI was higher in 1BL/1RS hybrids than in non-1BL/1RS hybrids; (2) 4 alien cytoplasms had positive effects on the FCU at MI in non 1BL/1RS hybrids; (3) The effect of 1B.1BL/1RS hetero-nucleus on meiosis was more obvious than that of alien cytoplasms in 1BL/1RS hybrids; (4) The selfed seed set of 1BL/1RS hybrids was not correlated to FCU, but was negatively correlated to FCLB; (5) The chromosomes' deeds at meiosis was more steady in non-1BL/1RS hybrids than in 1BL/1RS hybrids. The non-1BL/RS hybrids were easy to restore and restoring degree was high, and they had good prospects to be used. PMID- 11480178 TI - Distribution of donor genome composition and its application in marker aided backcrossing. AB - Conditional distribution was derived for the genome composition given the marker genotype of individuals from a backcross population. Genome size from donor was predicted by the mean. The precision of the prediction, indicated by the correlation with the unknown actual genome size from donor, was formulated by the variance and expressed as a function of the marker density. Results indicated that although high correlation can be obtained with a moderate marker density, say 40 cM per marker, the best individual in a large population has to be identified through a map of high density, say 10 to 20 cM per marker. For the best use of marker information in a marker aided backcrossing program for gene introgression, a stepwise marker genotyping and individual selection were suggested. That is, a preliminary selection can be made among all individuals but only on a few markers for each individual, and a fine selection would then be made among a few individuals but on more markers for each individual. This would be a very efficient method to increase the selection intensity in a large population but with a manageable number of marker genotyping for a gene introgression experiment. PMID- 11480179 TI - [Application of isonucleus and isocytoplasmic lines in the study of maize CMS]. AB - Between wild fertile type (F) and its sterile mutant (cms), if their nucleus and cytoplasm are the same, this wild fertile type (F) and its sterile mutant (cms) are called isonucleus and isocytoplasmic lines. The maize mtDNAs of isonucleus and isocytoplasmic lines (I), wild fertile type 478(F) and its sterile mutant 478 cms, were analyzed by RAPD. 94 primers were screened, 3 polymorphic products, OPZ 19(420), OPAA-15(600) and OPS-01(400), were amplified between 478(F) and 478-cms. The results showed that mtDNAs in isonucleus and isocytoplasmic lines were more homologous than that in others. There is minor mtDNA difference between the fertile type (F) and its sterile type (cms) in a pair of isonucleus and isocytoplasmic lines. The polymorphism detected in isonucleus and isocytoplasmic lines may be more closely linked with the gene of fertility. Therefore, isonucleus and isocytoplasmic line is an excellent system in the study of CMS. Sister isonucleus and isocytoplasmic lines are consisted of 2 groups of isonucleus and isocytoplasmic lines in which their nucleus are not all the same but closely related, their cytoplasm are the same. Using sister isonucleus and isocytoplasmic lines is equal to determining the change of fertility by transferring one cytoplasm into the isonucleus. Isonucleus and isocytoplasmic lines (II) are consisted of Su478(F) and Su478-cms. Isonucleus and isocytoplasmic lines (I) and (II) are sister isonucleus and isocytoplasmic lines. The polymorphic products, OPZ-19(420) and OPAA-15(600), can also be obtained in isonucleus and isocytoplasmic lines (II). The 2 polymorphic products OPZ-19(420) and OPAA-15(600) are existed in both of the isonucleus and isocytoplasmic lines. This showed that isonucleus and isocytoplasmic lines are practicable in the study of CMS, and that common polymorphism in isonucleus and isocytoplasmic lines may be related more directly to fertility. PMID- 11480180 TI - [Investigation of different dosages of inserted Bt genes and their insect resistance in transgenic Bt cotton]. AB - The plants with various dosages of inserted Bt genes were developed by intercrossing among transgenic Bt lines in which Bt genes were inserted in different chromosomes, and commercial non-transgenic Bt varieties. Laboratory bioassay of insect-resistance and Bt toxin protein assays with ELISA were used to detect efficacy to bollworm in different cotton growing stages. As transgenic Bt insect-resistant cotton, temporal difference of resistance existed in plants converging on different dosages of inserted Bt genes, i.e., there was a declining level of efficacy with plant age as the mortality (%) of Helicoverpa and Bt toxin protein expressional level decreased gradually, when analyzed with leaves from the main stem. Our research also revealed that the insect-resistance of converging 4 Bt genes plants was decreased due to homozygous suppression. PMID- 11480181 TI - [An AFLP marker related to fibrogenesis in upland cotton (Gossypium hirsuturm L.)]. AB - By using AFLP technique polymorphism analysis was performed between a fuzzlesslintless mutant line and its isogenic wild-type line, Xuzhou 142. Out of 6,360 bands produced by 64 pairs primers, a fragment, named as CF1, appearing stably in wild-type line, Xuzhou 142. This polymorphism was further verified using several normal fiber varieties and F2, F3 populations from the cross of fuzzless-lintless mutant line with a high-lint-percentage variety Yumian No. 1. The cosegregation of CF1 and fibrogenesis was proved, which suggested that CF1 can be used as a molecular marker for cotton fibrogenesis. The CF1 segment was cloned into PUCm-T Vector and then sequenced. The putative amino acid sequences, is an analogue to phenol hydroxylase alpha subunit, outer surface protein C, NADH dehydrogenase subunit 1, NADH-ubiquinone oxidoreductase, 2-oxoacid ferredoxin oxidoreduct and hypothetical 14.5kD protein. PMID- 11480182 TI - [Genetic diversity evaluation of cultivars (G. hirsumtum L.) from the Changjiang river valley and Tellow river valley by RAPD markers]. AB - Forty-one polymorphic decamer primers with clear and repeated band pattern, which were screened out from 300 randomly selected primers, were applied to assess genetic diversity of cultivars (G. hirsumtum L.) from the Changjiang river valley and the Yellow river valley by RAPD analysis. Eighty-four polymorphic loci were obtained from 84 cultivars from the Changjiang river valley and 96 polymorphic loci were obtained from 68 cultivars from the Yellow river valley. Jaccard's genetic similarity coefficients were calculated using the software of NTSYS-pc version 1.80, and two dendrograms were constructed by the unweighted pair group method of arithmetic average (UPGMA). Mean similarity coefficient among cultivars from the Changjiang river valley was up to 0.631. And cultivars from the Yellow river valley had an average similarity coefficient of 0.632. Compared similarity coefficient matrices and frequency distribution of pairwise similarity coefficients, a conclusion was drawn that cultivars from the two areas had nearly the same level of genetic diversity. Same base germplasm, breeding objectives, breeding methods and strategy could be attributed to similar genetic diversity between the two areas. PMID- 11480183 TI - Management of patients with severe oral mucosal disease. PMID- 11480184 TI - Advances in imaging in oral medicine. PMID- 11480185 TI - Problem solving in oral cancer. PMID- 11480186 TI - Current concepts in the management of patients with human immunodeficiency virus/acquired immune deficiency syndrome. PMID- 11480187 TI - Systematic approach for treating the medically complex dental patient. PMID- 11480188 TI - Geriatric oral medicine. PMID- 11480189 TI - [The pediatric surgeon and injuries]. PMID- 11480190 TI - [Progress in laryngotracheal surgery]. AB - The number of patients with laryngotracheal disorders is small but they have great importance because they produce anxiety in parents and they have a considerable morbi-mortality rate. We present the state of the art in three groups of patients: subglottic stenosis, tracheobroncomalacia and tracheal stenosis. PMID- 11480191 TI - [Evaluation of thoroughness and legibility of informed consent documents in pediatric surgery]. AB - BACKGROUND: The written information that is provided to the patient must gather a series of characteristics, quantitative and qualitative, so that fulfill the ethical criteria that govern the theory of the Informed Consent. AIMS: To evaluated the degree of exhaustivity and readability of a series of Informed Consents forms in Pediatric Surgery units. MATERIAL AND METHODS: 1. Forms: We evaluated 118 consent forms submitted by 17 pediatric surgery units. 2. Exhaustivity: The studied variables have been: name of the hospital, name of the patient, name of the physician, date, description of the procedure, objectives, risks mention, personalized risks, possibility of recovery, benefits of the procedure, alternatives, possibility from withdrawing the assent, arrangement to widen the information, comprehension of the document, express assent and signatures. All the variables are dichotomics, being collected your presence or absence. 3. Readability: Each document has been introduced in the program "Corel Word Perfect". It was applied to the usefulness "Grammatik" and were collected: number of words, number of phrases, Flesch/Kincaid index, Orational Complexity index and LEGIN index. RESULTS: 1. Exhaustivity: The paragraphs presence in the different forms by order of frequency were: physician signature (100%), express assent (100%), signatures (100%), date (98%), name of the patient (97%), comprehension of the forms (97%), name of the physician (94%), risks mention (92%), name of the hospital (85%), description of the procedure (78%), possibility from withdrawing the assent (70%), personalized risks (67%), arrangement to widen information (43%), to explain alternatives (41%), objectives of the procedure (36%), benefits (36%), and possibility of the recovery (16%). 2. Readability: 11.1% of the forms fulfills the Flesch index, 27.9% the Orational Complexity index, and 22.1% LEGIN index. CONCLUSIONS: Most of the documents offer sufficient quantity of information with respect to the formal aspect and the risks an less with respect to the benefits. This seems to indicate your defensive character. Equally, they are difficult to reading and by difficult to understanding. PMID- 11480192 TI - [Bacterial translocation associated with short bowel: role of ileocecal valve and cecum]. AB - Sepsis in short bowel syndrome (SBS) is due in part to bacterial translocation (BT). Parenteral nutrition (PN) is often necessary in SBS and promotes BT. The presence of ileocecal valve (ICV) has been considered as a good prognostic factor in the outcome of this children. The aim of this study was to asses the effect of the presence or absence of ICV and cecum in five different models of gut resection in the rat. Fifty-five adult Wistar rats were randomly assigned to one of five groups: Group 1 (N = 14): standard rat chow + 80% small bowel resection. Group 2 (N = 10): standard rat chow + 80% small bowel resection including cecum. Group 3 (N = 10): standard rat chow + 80% small bowel resection including ICV. Group 4 (N = 11): NP + 80% small bowel resection. Group 5 (N = 10): NP + 80% small bowel resection including ICV and cecum. Ten days after surgery they were sacrificed and mesenteric lymph nodes (MLN), spleen and peripheral (PBL) and portal blood (POBL) specimens were recovered and cultured. Groups 3 (without ICV, with cecum) and 5 (without ICV, without cecum) showed 60% BT in MLN and POBL, and groups 1 and 4 (with ICV, without cecum) 93% and 91% respectively (p < 0.05). In PBL, group 3 (without ICV, with cecum) showed also less BT than groups 1 and 4 (10% vs 43% and 55% respectively, p < 0.05) and group 5 (without ICV and cecum) had less BT than groups 1, 2 and 4 (0% vs 43%, 30% and 55%, p < 0.01). In conclusion, these results suggest that the absence of ICV decreases BT and that the cecum does not seems to play a role on his. PMID- 11480193 TI - [Gastric tonometry as monitoring system in abdominal surgery]. AB - OBJECTIVE: The aim of this study is the evolution of the intramucosal pH (pHi) during the quirurgic and the immediate postoperative process in pediatric patients, operated in abdominal surgery longer than 90 minutes. MATERIAL AND METHODS: A prospective study including 25 patients was done. In all of them intraabdominal surgery was performed. The mean age was 5.6 +/- 3 years, range from 9 months to 13 years. The operative time was 207 +/- 76 minutes. The pHi was measured by means of a sigmoid tonometer introduced in the stomach. A tonometry was done hourly during the operative procedure, and every 8 hours during the postoperative period. RESULTS: 157 determinations of pHi were made with an average of 6.28 per patient. During the operative period the pHi decreased from 7.36 +/- 0.11 (30') to 7.22 +/- 0.11 (210'). During the postoperative, the pHi increased to 7.36 +/- 0.05, 24 hours later (p < 0.005). CONCLUSIONS: During the surgery there was a significant reduction of the pHi, which recovered 16 hours after in the postoperative. PMID- 11480194 TI - [Prognosis in children with biliary atresia successfully treated with Kasai's operation]. AB - BACKGROUND: The Kasai procedure, portoenteroanastomosis (PEA) didn't reach international spreading until the seventy's decade, making difficult to find long term results from children with ABE successfully treated with this technique. At our institution in the last fifteen years all the therapeutics procedures for these patients can be offered, including the liver transplant. AIM: To show the evolution of our patients with ABE treated with the PEA and that survive long term without being transplanted. METHODS: The clinical course of 22 patients that survive more than 10 years after the PEA with their own liver is reviewed. The hepatic survival indexes of (success, death or transplant) are beyond the tenth year. The problems raised during the follow-up are analysed. RESULTS: From 99 patients with ABE treated primarily in our center, 22 reached the 10 year-old age after the PEA without a liver transplant. In the follow-up, seven if the these finally needed the transplant. Their median age was 12.2 year-old (range: 10.5 13.8) for a progressive hepatocellular damage in 5 cases associated to syndrome hepatopulmonar in two cases. The other fifteen patients have a compensated hepatopathy. Five of them do not have hyperesplenisme and the serum bilirrubine levels are lower than 1.3 mg/dL. The medium age of these patients at the end of the follow-up was 14.8 years. CONCLUSIONS: In spite of the reestablishment of the biliary flow with the PEA, few are the patients with ABE that preserve their hepatic function lapsed long periods of time. Nevertheless the prognosis of these patients is excellent. PMID- 11480195 TI - [Laparoscopic splenectomy in the treatment of hematologic diseases]. AB - Laparoscopic approach to surgical procedures in children is used in different diseases, but there are some controversies for using this technique in the splenectomy. Laparoscopic surgery decreased pain, postoperative ileus and the hospital stay; and improve cosmetics, the most important disadvantage is the longer operating time. Splenectomy is indicated more in immune thrombocytopenic purpura (PTI) and hereditary espherocytosis (HE). We review our experience in laparoscopic spelenectomy. PMID- 11480196 TI - [Laparoscopic splenectomy in pediatric hematologic diseases]. AB - INTRODUCTION: Surgical acceptance of the laparoscopic splenectomy in the pediatric hematological disorders has improved due to safety, to relif postoperative pain, and to reduce the hospital stay. The aim of this study is present our surgical laparoscopic technique performed. MATERIAL AND METHODS: Eight patients, 6 spherocytosis (age ranged: 6-14 years, average age: 9.5 years; 5 girls and 1 boy) and 2 cases of autoimmune hemolytic anemia (both boys of 18 months and 4 years) underwent laparoscopic splenectomy. The Trias-Targarona technique has been performed placing the patient at the right lateral-decubitus position. Four trocars are placed, two of 5 mm in the posterior axillary line and epigastrium and other 2 trocars of 12 mm, in the middle of them. RESULTS: In all these 8 patients a laparoscopic splenectomy was completely performed. We had 2 cases with problems with the sac introduction. Cosmetic results have been excellent and we changed the 7-day antibiotic protocol for 2-day protocol. The surgical time has changed from 3 to 2 hours. CONCLUSIONS: With laparoscopic splenectomy we can give an adequate response to hematological disorders in children with a acceptable surgical time, an excellent cosmetic result with a minimum postoperative time and a very short hospital stay. PMID- 11480197 TI - [Colonic aganglionosis with ileal involvement. Ileocolic plasty]. AB - During 1994 through 1999, we have treated five patients (3 boys, 2 girls) with total colonic aganglionosis (TCA) and ileal involvement. In three of them we performed a diverting ileostomy in the neonatal period and at the age of four and five months respectively in the remaining two patients, due to intestinal obstruction. In these two last patients a diagnosis of Hirschsprung's disease was made by anorectal manometry and rectal biopsies in the neonatal period. During laparotomy, a cutaneous ileostomy was created in all patients at the distal end of normal ileum, which was 30 to 110 cm (mean = 71 +/- 37 cm) from the ileocecal valve. After operation, a short bowel syndrome developed in three patients causing fluid and nutritional problems that required prolonged total parenteral nutrition (TPN). The definitive operative repair is performed at 4.5 to 14 months (mean = 8 +/- 3.7 months) by a 12 to 20 cm side-to-side ileocolostomy created between the ileum and aganglionic ascending color (Boley procedure) and ileorectal primary anastomosis (Rehbein procedure) using a circular stapler. Rectal dilatation, irrigation of the colon with saline, loperamide hydrochloride and resincholestyramine were begun in all patients postoperatively. Oral antibiotic prophylaxis, was given to the three patients who suffered from SBS. Oral feedings with semielemental diet were tolerated once stools were semiformed and TPN was discontinued at 8 to 34 days (mean = 21 +/- 1.7 days). postoperatively. After the definitive operation, enterocolitis developed in two patients, requiring one of them a short hospitalization during the episode. A possible explanation for the low incidence of enterocolitis in this series is the systematic postoperative use of irrigations of the colon with saline in all patients. These five patients have been followed-up for growth, development, bowel habit and continence. Follow-up has ranged from 15 to 62 months (mean = 32.2 +/- 19.2 months). Presently, all patients in this series have full enteral feeding and one to three bowel movements per day, with formed or semiformed stools. No patients is incontinent of stool. The patient's body weights (74 to 93%) and heights (89 to 92%) for their age were below average (four patients) or within normal range (one patient). Hemoglobin levels are within normal range in all patients. One patient has iron deficiency and another one has serum ferritin concentration below normal. Three patients have folic acid deficiency. Vitamin B12 absorption is normal in all patients. Although we can not conclude this is a better procedure than others, with is use we have obtained satisfactory results, with an excellent survival, scanty morbidity, a rapid return of bowel function and continence, and an acceptable physical development. During long-term follow up, patients mus be evaluated for iron and folic acid deficiency. PMID- 11480198 TI - [Gastric bezoar]. AB - Bezoar consist of the accumulation of different substances in the digestive tract. We present the clinical case of a girl with gastric trichobezoar discovered in a postoperatory follow-up to an appendicectomy. Ultra-sound images and oesophagus-gastric transit are fundamental points in the diagnosis. Endoscopy was useful in confirming the diagnosis and in ruling out associated ulcus, frequent in these patients. Surgical treatment is the choice in big bezoar but laser, prokinetics and enzymatic dissolutions have also been used. Theses patients should undergo psychological control. PMID- 11480199 TI - [Results evaluations 3 years after our first experience with the laparoscopic treatment of Hirschsprung disease]. AB - The laparoscopic approach has been successfully used during the recent last years in pediatric surgery. We describe the first experience in Argentina using laparoscopic surgery in November 4th in 1996, for the treatment of a Hirschsprung disease case. We used the technique of Soave's modified by Georgenson. Details of the surgical procedure are given. Hospitalization time took 5 days and immediated postoperatory cares were well done. Soave-Georgeson technique seems to be a safe and effective possibility for the Hirschsprung disease treatment, and it is completely achieveable by laparoscopy. The present experience does not show advantages in terms of a lower hospitalization time, however it really shows the benefits of laparoscopic surgery, such as post-operatory comfort and better cosmetic results, both highly superior than those obtained with classical surgery. PMID- 11480200 TI - Improving inpatient antibiotic prescribing: insights from participation in a national collaborative. AB - BACKGROUND: Increasing rates of antimicrobial resistance and the role antibiotic overuse plays in contributing to this problem have been widely documented and have prompted appeals to change prescribing practices. How to actually achieve such changes represents a major challenge. As part of the Institute for Healthcare Improvement (IHI) Breakthrough Series project Improving Prescribing Practices (IPP), in 1997-1998, Cook County Hospital (Chicago) worked with other institutions that chose antibiotics as their focus in this national collaborative. PRACTICAL SUGGESTIONS: Practical suggestions are offered within six categories--adopting a general approach to improving antibiotic prescribing (marshalling credible evidence, addressing physician concerns and skepticism, and removing barriers to make it easier for prescribers to change); rethinking guidelines (providing syndrome-based guidance, revising national guidelines for local use, defining scenarios in which drug(s) can be safely withheld, offering alternatives, prospectively resolving conflicts over drug of choice and empiric regimens, and defining situations in which immediate treatment is and is not critical); getting the message out and changes implemented (using antibiotic order forms, computer order entry, and infectious disease specialist consultation); building viable linkages to leverage change (bridging disciplines); improving measurement; and promoting nondrug strategies and patients' roles in treating and preventing infection. CONCLUSION: Antimicrobials are unique, being the only class of drug therapy that affects not only the patient to whom it is prescribed but other current and future patients as well. Institutions therefore have a special responsibility to ensure their efficient and judicious use. It is often easier to prescribe antibiotics than to exercise restraint. PMID- 11480201 TI - The search for good antimicrobial stewardship. PMID- 11480202 TI - Designing an intervention to improve the management of Helicobacter pylori infection. AB - BACKGROUND: An intervention to improve the testing and treatment of Helicobacter pylori (HP) in patients receiving chronic acid suppression (AS) therapy was developed at Harvard Pilgrim Health Care (HPHC), a mixed-model not-for-profit health maintenance organization. METHODS: Ten full-time primary care physicians (4 staff model and 6 group practice) were interviewed in 1999 about their knowledge, attitudes, and practice regarding dyspepsia, the use of chronic AS drugs, and approaches to HP infection, as well as about the feasibility and acceptability of various potential interventions that might be used in a quality improvement program. RESULTS: Self-reported practice regarding dyspepsia and HP infection were relatively uniform, and physicians were generally aware of current recommendations. Three common misperceptions acted as barriers to optimal HP management: Untreated HP was not considered an important problem; patients who used drugs for chronic AS rarely had HP infection; and chronic use of AS drugs was considered effective and without adverse consequence. All physicians wanted brief educational materials with explicit guidelines, preferably locally adapted and endorsed by local experts. All informants agreed that the main barrier to successful QI interventions was the requirement for any extra time or effort, particularly when directed at populations of patients who do not have symptomatic complaints. DISCUSSION: The interviews revealed the many barriers to improving the management of HP infection and to targeting educational messages and tailoring different methods for facilitating practice change across different managed care settings. Evidence-based components of the intervention program include physician education, a notification/reminder system, and practice-based tools to facilitate change and minimize workload. PMID- 11480203 TI - Measuring and improving patients' and families' perceptions of care in a system of pediatric hospitals. AB - BACKGROUND: Shriners Hospitals for Children (SHC) is a network of 22 pediatric specialty hospitals that provide medical care free of charge to children up to 18 years of age and that serve as referral centers for children with complex orthopedic and burn problems. In 1998 the SHC system began using The Picker Institute's Patient and Family Perception of Care inpatient survey throughout its hospitals. SYSTEMWIDE IMPLEMENTATION: A broad-based implementation plan was developed to promote acceptance of the perception of care topic and provide education on performance improvement. In 1999 a work group was formed to prioritize areas for improvement, survey benchmark hospitals, and identify best practices in benchmark hospitals. This work group first focused on the dimensions of Partnership Between Families and Clinicians and Information and Education to the Child. In May 1999 the work group began the task of identifying best practices in these two priority dimensions from the SHC benchmark hospitals. Surveys were submitted to those hospitals, asking what they perceived as being the reasons they scored well in those areas. The results of these surveys were used to identify key practices in these benchmark hospitals that are of significant importance in patient and family perceptions of quality care. NEXT STEPS: The challenge is to facilitate cross-facility interactions to understand and adopt best practices. Focus groups will be conducted to further delineate the dimensions with higher problem scores. The SHC system plans to expand the patient surveys to outpatients, to allow for the evaluation of the full complement of hospital patients. PMID- 11480204 TI - Promoting patient safety: is technology the solution? AB - BACKGROUND: On April 30, 2001, the Cleveland Clinic Foundation and Cleveland Clinic Health System Quality Institute sponsored a 1-day conference focused on technology in patient safety. PATIENT SAFETY-A CALL TO ACTION: Kenneth W. Kizer focused on ten high-priority patient safety strategies identified by the National Quality Forum-including implementing recognized "safe practices", recognizing and dealing with professional misconduct, and supporting efforts to create a nonpunitive environment for health care error reporting. CULTURAL IMPLICATIONS OF INTRODUCING NEW TECHNOLOGY: Randolph A. Miller described a computerized clinician order-entry system used to provide decision support, reduce excess test ordering, introduce cost savings, and meet regulations for inpatient radiology and cardiology tests. USING BAR CODES TO ELIMINATE MEDICATION ERRORS: Jeff Ramirez reported on the Veterans Health Administration's use of bar coding technology for point-of-care validation of medication administration, which has resulted in improvements in response time; the efficiency of the dispensing, delivery, and administration process; and patient care. HOW TO MAKE COMPUTERS TEAM PLAYERS: The knowledge base exists to design computers as team players that expand human expertise and help health care practitioners better create safety. Yet David D. Woods challenged the audience to anticipate the changing shape of iatrogenic risk as a result of increasing dependence on automation in health care. TECHNOLOGY AND MEDICATION SYSTEMS: Mark Neuenschwander spoke about automating various steps within the medication use system, through computerized prescriber order entry and bedside scanning. FUTURE TECHNOLOGICAL POSSIBILITIES: Charles Denham suggested how technology may aid health care professionals in their care of patients, such as in using predictive modeling to identify the risks of therapeutic intervention. PMID- 11480205 TI - Absorption and accumulation of nitrate in plants: influence of environmental factors. AB - Plants adopt various strategies to fulfill their nitrogen nutrition requirement, the most important being the uptake of nitrate from the soil and its subsequent assimilation in to amino acids. The uptake of nitrate is energy dependent and is an active process involving high affinity and low affinity transport systems. The net uptake of the anion depends upon both influx as well as on its passive efflux. When the uptake far exceeds over its assimilation in the plant, there is considerable accumulation of nitrate in the plant parts making them unfit for human and cattle consumption. Various environmental factors affect the uptake and accumulation of nitrate, which along with the genetic component of the plant affecting the net uptake and accumulation of the nitrate, need to be considered and carefully manipulated for effective nitrogen management in the plant, soil and aquatic environment. PMID- 11480206 TI - Induction of lymphomas on implantation of human oral squamous cell carcinomas in nude mice. AB - Cancer cells from five oral cancer patients and pleomorphic adenoma cells from one individual were inoculated as single cell suspension into subcutis of 30 Swiss nude mice and tail vein of additional 30 mice. Further, tumor tissue pieces from three oral cancer patients were xenografted s.c. in 18 nude mice, and 10 mice were kept as controls. In animals implanted with tumor pieces, 7/18 (39%) mice, developed squamous cell carcinoma at the site of inoculation within 8-15 days, while tumors were not observed in mice inoculated with single cell suspension, up to 60/90 days. In 8/68 (12%) mice, white foci were observed in several tissues, with hepatomegaly and splenomegaly noted in 27/68 (39%) mice. Histopathological examination of various tissues revealed presence of large cell lymphoma in several organs in 14/68 (21%) mice. No regional or distant metastasis of the implanted oral tumor cells was detected. Mice injected with cells from pleomorphic adenoma, also demonstrated large cell lymphoma in 2/10 (20%) mice, whereas none of the 10 control animals showed any gross abnormalities or microscopic abnormalities in several organs. 2/16 (12%) lymphomas exhibited positive reaction with mouse B cell antibodies illustrating the murine origin of the lymphomas, and these were immunophenotyed as B cell lymphomas. The lymphomas were also examined with mouse T cell antibodies and none reacted positively with the mouse T cell antibodies. The lymphomas also failed to react with human T cell, B cell and human Leucocyte common antigen (LCA) antibodies, indicating that the induced lymphomas were not of human origin. The tumor specimens from seven of eight oral cancer patients and the pleomorphic adenoma patient induced lymphomas in nude mice. Thus it appears that xenografting oral tumor cells into nude mice may cause induction of the murine lymphomas, and this needs further investigation. PMID- 11480207 TI - Fine structure of prolactin cell of female albino rat as affected by some antifertility drugs--a comparative electron microscopic study. AB - One antioestrogenic compound as well as some antifertility drugs have been administered to female albino rats over a period of six months to study their long term effects on fine structures in PRL cell. Almost in all the cases, the dynamics of hormone synthesis and secretion have been affected. Fine structure is suggestive of activation of synthetic machinery of the cell. The cell picture under the estradiol valerate regimen presents a transitional stage progressing towards involution due to accelerated cell cycle. Sparse granulation, frequent granule extrusion and misplaced exocytosis under the influence of tamoxifen citrate or levonorgestrel + ethinyloestradiol are similar to those observed in adenomatous PRL cell. Fine structural correlates of stepped up synthesis are also observed following chronic progesterogenic influences of progesterone and norethisterone heptanoate, but the magnitude of the change is on a lower scale. All the fine structural changes have been discussed in the context of ultrastructural pathology. PMID- 11480208 TI - Modulatory effect of quercetin on azathioprine induced membrane changes in the mouse spleen. AB - Modulatory effect of quercetin on azathioprine induced toxic changes was studied in spleen of experimental animals. Azathioprine treatment caused an increase in serum albumin/globin ratio and a decrease in total protein in spleen tissue. An increase in a membrane bound ATPases was also noted. Supplementation of quercetin with azathioprine increased the protein content and lowered the activities of membrane ATPase in spleen. There was a decrease in serum albumin globulin ratio. It was concluded that quercetin modulated the protein and membrane bound ATPase activities and protected the spleen from azathioprine induced membrane damage. PMID- 11480209 TI - Effect of Centella asiatica Linn on physical and chemical factors induced gastric ulceration and secretion in rats. AB - Centella asiatica is commonly mentioned as a Rasayana in Ayurveda, an ancient system of Indian medicine for various ailments including abdominal disorders. Rasayanas have been advocated for use in rejuvenation therapy. The present study was conducted to evaluate the possible anti-ulcerogenic activity of fresh juice of C. asiatica (CAJ) against ethanol-, aspirin-, cold-restraint stress- and pyloric ligation induced gastric ulcers in rats. The drug given orally in doses of 200 and 600 mg/kg twice daily for five days, showed significant protection against all the above experimental ulcer models and the results were comparable with those elicited by sucralfate (SF, 250 mg/kg, p.o., BD x 5 days). CAJ showed little or no effect on offensive acid-pepsin secretion. However, at 600 mg/kg CAJ significantly increased gastric juice mucin secretion and increased the mucosal cell glycoproteins signifying increase in cellular mucus. It also decreased cell shedding indicating fortification of mucosal barrier. Thus, the ulcer protective effect of CAJ may be due to strengthening of the mucosal defensive factors. PMID- 11480210 TI - Host-mediated juvenoids affect development and metamorphosis of endoparasitic wasp, Chelonus blackburni (Hymenoptera: Braconidae) and consequent morphogenetic derangements in its reproductive system. AB - Juvenoids, applied topically on larvae of Corcyra cephalonica harbouring the larvae of the parasitoid, produce various types of developmental derangements in parasitoid C. blackburni. The deformed morphs, e.g., apparently normal adults, adultoids and prolonged larvae were developed at different rates by different doses of juvenoids. Larval and pupal mortality rates of the parasite were 6-13 and 4-8% respectively in different treatments. Deformities in reproductive system of the parasitoid were increased or decreased ovariole number, development of compound egg chamber and ill-developed ovariole. PMID- 11480211 TI - Screening of Bacillus thuringiensis serotypes by polymerase chain reaction (PCR) for insecticidal crystal genes toxic against coffee berry borer. AB - Using PCR,257 isolates of Bacillus thuringiensis(Bt) were screened for cry-type genes. Of 257 isolates/strains, 60 isolates were identified as cry7/8, 10 isolates as cry3 and 36 isolates as cry 1I. One specific strain of B. thuringiensis (sumiyoshiensis; T03B 001) was investigated for the presence of cry7 and cry8 genes. Genes Cry7 and cry8 were first detected in this strain using family primers prior to analysis by exclusion polymerase chain reaction (E-PCR) using specific type primers. E-PCR conducted with the above said primers led to the identification by agarose gel electrophoresis of a remaining 1.5 Kb family band indicating a potentially novel gene. This PCR product, (1.5 Kb), was purified from the gel and cloned in pGEM-T Easy vector. Twenty recombinant colonies bearing 1.5 Kb insert were identified and three randomly selected representatives of the group, clones 7, 8 and 10, were sequenced and compared to all cry7 and cry8 sequences available from Gene Bank. Alignments with available DNA and protein sequences showed that all these clones contained a gene related to cry8Aa1. Analysis using protein sequence alignment showed that the sequence from clone 7 differed from the closest relative, known under the new nomenclature as cry 8Aa1, by 44%. The crystal proteins from B. thuringiensis sumiyoshiensis (T03B 001) was toxic to coffee berry borer larvae. PMID- 11480212 TI - Extracellular polysaccharide production by Azorhizobium caulinodans from stem nodules of leguminous emergent hydrophyte Aeschynomene aspera. AB - The Azorhizobium caulinodans isolated from the stem nodules of a leguminous emergent hydrophyte, Aeschynomene aspera, produced a large amount of extracellular polysaccharides (EPS) in yeast extract basal medium. Maximum EPS production was at the stationary phase of growth. EPS production was increased by 919% over control when the medium was supplemented with sucrose (1.5%), D-biotin (1 microgram/ml) and casamino acid (0.1%). EPS contained rhamnose and arabinose. Possible role of the azorhizobial EPS production in the stem nodule symbiosis is discussed. PMID- 11480213 TI - Association of lignifying enzymes in shell synthesis of oil palm fruit (Elaeis guineensis--dura variety). AB - Scanning electron microscopic (SEM) observation demonstrates the differentiation of mesocarp and endocarp tissues and their lignified nature in dura fruits at 8 weeks after pollination (WAP). During shell formation, the endocarp cells become lignified to a hard shell while the mesocarp tissue remains cellular and fibrous. A transition zone made up of fibrous units was also visible beneath the shell. The soluble phenols of mesocarp and endocarp tissues at their developmental stage was analyzed using Reverse phase high performance liquid chromatography (RP HPLC). The appearance of ferulic acid at 4 WAP and its absence at 8 WAP indicates the role of ferulic acid in lignin synthesis. The HPLC data was supported by the lignin concentration. To ascertain the biochemical relationship of lignin pathway enzymes, phenylalanine ammonia lyase (PAL), cinnamyl alcohol-NADPH-dehydrogenase (CAD) and peroxidase (POD) with shell synthesis, the activities of these enzymes and lignin content were assessed during development of the shell between 4 and 8 WAP. The three enzymes, PAL, CAD and POD expressed high level of activity in the mesocarp and endocarp at 4 WAP. At 8 WAP a sharp decline in activity was observed in the endocarp whereas the mesocarp showed a moderate reduction. This variation is an indication of the role of these enzymes in shell formation. PMID- 11480214 TI - Augmenting in vitro shoot multiplication by vipul (triacontanol) and adventitious rhizogenesis by rice bran extract in Dendrocalamus strictus. AB - Like other bamboo species, Dendrocalamus strictus flowers gregariously after a prolonged intermast period of 48 years and constitutes an ideal material for in vitro clonal propagation. In this study, MS liquid medium containing 0.5, 1.0 and 2.0 mL/L vipul (Godrej Agrovet, Ltd., Sachin, India), a commercial formulation of triacontanol, with or without BA (3.0 mg/L) was tested for in vitro shoot multiplication and 1.0, 2.5 and 5.0 mL/L of 20% (w/v) alcoholic/aqueous rice bran extract (alone or in combination) with NAA (3 mg/L) used for in vitro adventitious rhizogenesis in single node culture derived shoots of Dendrocalamus strictus.. After a multiplication cycle for 4-5 week, vipul (0.5 mL/L) with BA (3.0 mg/L) in the culture medium induced 4.59 fold shoot multiplication rate whereas application of BA and vipul alone had corresponding values of 3.29 and 0.53 fold respectively. Maximum vipul concentration (2 mL/L) with BA (3 mg/L) exhibited shoot multiplication higher than (or equal to) that of BA alone. Maximum in vitro rooting percentage (55.66%) was obtained on half MS medium enriched with alcoholic rice bran extract (2.5 mL/L) and NAA (3 mg/L). This is the first investigation reporting amelioration of in vitro shoot multiplication rate by triacontanol and rooting percentage by rice bran extract in explants from mature bamboo culms. The protocol is economical and rapid for in vitro clonal propagation of Dendrocalamus strictus. PMID- 11480215 TI - Ultrastructural changes in testis of gobiid fish Glossogobius giuris (Ham) induced by fenthion. AB - Pesticides are known to affect the testis in fishes and cause cytomorphological changes or gross physiological alterations due to changes in its histology, retard gonadotrophic secretions thereby causing regression of the gonads. Light and ultrastructural studies were made on control and fenthion treated testis, interstitial gland and sperms of G. giuris during spawning phase after exposing them to different sub-lethal concentrations (0.05, 0.25 and 0.5 ppm) fenthion for a short-term period (24, 48, 72 and 96 hr). The results indicated extensive cytotoxic damages in the testis including atrophy of testis, reduction in lipoidal material in the interstitial cell and degeneration of sperms. The alteration in histology of testis, interstitial cell and sperms progressively increased with the increase in the sub-lethal concentration of pesticides. PMID- 11480216 TI - Chlorpyrifos induced alterations in levels of thiobarbituric acid reactive substances and glutathione in rat brain. AB - Chlorpyrifos, O,O'-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothionate, exposure in rats caused significant inhibition of acetylcholinesterae activity in different regions of brain and generated oxidative stress as evidenced by increase in the level of thiobarbituric acid reactive substances and decrease in the ratio of reduced to oxidized glutathione in all the three regions of brain. Malondialdehyde level was increased significantly in all regions of brain and the increase was dose dependent. Mid brain showed highest level of lipid peroxidation. PMID- 11480217 TI - Prevention of adrenocortical hyperactivity by dietary casein in rats exposed to forced swimming stress. AB - Adrenal weight, adrenal hydroxysteroid dehydrogenase activity and serum corticosterone level were significantly higher in rats fed with 5% casein diet after 7 days of swimming stress (45 min/day) as compared to their controls. All the parameters were similar to their control levels in rats receiving 20% casein diet and exposed to swimming stress. The results suggest that casein can play an important role in preventing adrenocortical hyperactivity in swimming stressed rats. PMID- 11480218 TI - Effect of Indigofera tinctoria Linn on liver antioxidant defense system during D galactosamine/endotoxin-induced acute hepatitis in rodents. AB - Effects of pre-treatment with the alcoholic extract of I. tinctoria (500 mg/kg body wt/day, p.o. for 21 days) on liver antioxidant defense system during acute hepatitis induced by D-galactosamine (D-GalN)/endotoxin (LPS extracted by phenol water method from E. coli serotype 0111.B4; 300 mg and 30 micrograms/kg body wt/day, i.p., 18 hr before the assay) were investigated on the activities of enzymic antioxidants such as superoxide dismutase, catalase, glutathione peroxidase and glutathione-s-transferase, and levels of total reduced glutathione in the liver of normal and experimental groups of male albino rats. Since lipid peroxidation and associated membrane damage is a key feature of D-galN/LPS induced liver injury, the levels of lipid peroxides, was estimated and used as an index of oxidative stress. D-GalN/endotoxin-induced hepatic damage was manifested by a significant decrease in the activities of antioxidant enzymes, decreased glutathione levels and increased levels of lipid peroxides. I. tinctoria pre treated rats showed considerable protection against D-galN/endotoxin, induced oxidative stress as evidenced by a significant increase in the activities of all the antioxidant enzymes studied and significant decrease in the levels of lipid peroxides. Results indicate that pretreatment with I. tinctoria extract in rats is very effective in reducing D-GalN/endotoxin-induced oxidative stress suggesting an antioxidant effect. PMID- 11480219 TI - Emergence of Mendelism: pages from history. PMID- 11480220 TI - Insights into the molecular basis of male infertility. AB - The study of the genetic basis of human male infertility is complicated by genetic heterogeneity and because linkage analysis studies are difficult. The study has been limited so far to the analysis of genes located on the Y chromosome. Several genes and gene families have been discovered and mutation analysis of these candidate genes in infertile patients is ongoing. In recent years, several mouse models with impaired spermatogenesis or fertility have also been analysed, expanding our knowledge about the molecular basis of spermatogenesis and male fertility. PMID- 11480221 TI - Animal models of tardive dyskinesia--a review. AB - Tardive dyskinesia is a serious motor side effect of long term neuroleptic therapy, with an unknown pathophysiological basis. The leading hypothesis of the pathophysiology of tardive dyskinesia includes dopamine receptor supersensitivity, GABAergic hypofunction, excitotoxicity and oxidative stress. Many preclinical models have been developed to identify the underlying pathological processes of tardive dyskinesia, but none has yet produced a parsimonious results. A wide range of animal models, viz. Homologous, analogous and correlational models have been developed to explore the pathophysiology of tardive dyskinesia. Vacuous chewing movements in rodents induced by chronic neuroleptic treatment is the most frequently employed model. As the existing models suffer from several phenomenological and methodological problems, development of new models, highly predictive of pathological basis of tardive dyskinesia can accelerate tardive dyskinesia research for the better understanding of the pathophysiological processes underlying the syndrome and for the discovery of new therapeutic targets for the treatment of tardive dyskinesia. PMID- 11480222 TI - Morphological and functional characteristics of rabbit uterine epithelial cells grown on free floating collagen gel. AB - In the present study isolated uterine epithelial cells from normal rabbits were maintained in culture on free floating rat-tail collagen matrix, and the morphological characteristics of these cells were examined. Additionally, the pattern of protein synthesis and secretion by rabbit uterine epithelial cells grown on free floating collagen gels following estradiol and/or progesterone treatment in vitro was examined. Isolated epithelial cells cultured on collagen gels in complete medium containing serum attached to form monlayers, and eventually the gels became free floating and contracted giving rise to luminal arrangements. These cells were cytokeratin positive epithelial cells and were ultrastructurally polarized. These cells also exhibited differential upregulation and down regulation in the synthesis and secretion of proteins in response to estradiol, progesterone, and estradiol plus progesterone. Additionally, a permissive action between progesterone and estradiol in the synthesis of two species of secretory proteins was observed. It however remains to be examined whether different species of proteins produced in vitro in response to estradiol and progesterone bear any association with physiological states in reproductive cycle in this species. PMID- 11480223 TI - Modulation of feeding and drinking behaviour by catecholamines injected into nucleus caudatus in rats. AB - Lesions of nucleus caudatus have been documented to produce adipsia and aphasia in rats. Injection of dopamine into this nucleus has been shown to facilitate water intake in rats. But, reports are not available on the effects of intracerebral injection of epinephrine and norepinephrine on feeding and drinking behaviour in animal models. Therefore, in the present study the effect of adrenaline and noradrenaline injected into nucleus caudatus on food and water intake in rats was assessed. 24 h basal food and water intakes were recorded in Wistar rats and were found to be 12.37 +/- 0.20 g and 22.04 +/- 0.27 ml respectively. Stainless steel cannulae were implanted stereotaxically into the nucleus caudatus. Four different doses (0.1 microgram, 0.5 microgram, 1 microgram, and 2 micrograms) of adrenaline and noradrenaline were injected into the nucleus caudatus through the implanted cannulae in separate groups of animals and their 24 h food and water intakes were recorded following these injections. No change in food and water intake was observed following the administration of different doses of adrenaline. A significant increase in 24 h food intake reaching a maximum of 16.03 +/- 0.15 g at 2 micrograms dose, without change in water intake was observed following administration of different doses of noradrenaline. The noradrenaline-facilitated food intake was blocked when noradrenaline was injected following injection of phentolamine, an alpha-receptor blocker. The bilateral lesions of nucleus caudatus resulted in a significant and sustained inhibition of food (8.98 +/- 0.17 g) and water intake (19.12 +/- 0.16 ml). These observations suggest that nucleus caudatus is involved in regulation of food and water intakes in rats. Noradrenaline-facilitated food intake is mediated by alpha-receptors. Adrenaline does not affect these ingestive behaviours when injected into the nucleus caudatus in rats. PMID- 11480224 TI - Abuse liability of diazepam through different routes. AB - Abuse liability of diazepam was examined among experienced drug users. The subjects, randomly divided into two groups, each having eight subjects, received diazepam, 15 mg intravenously (gr. 1) and 30 mg orally (gr. 2). Subjective states, drug liking, sedation and euphoria were assessed at pre-drug, 15 min, 120 min and 240 min post-drug administration. In addition, brief assessment to evaluate euphoria and sedation was carried out at 5 min and 45 min for subjects in gr. 1, and at 45 min and 150 min for those in gr. 2. Plasma diazepam level was also estimated. Results indicate those subjects in gr. 1 reported quicker and higher euphoria, drug liking, subjective effects and higher plasma level. The study suggests that route of administration of a compound which has faster onset of action is associated with more liability of abuse. PMID- 11480225 TI - Oncoprotein c-erbB-2 in squamous cell carcinoma of the uterine cervix and evaluation of its significance in response of disease to treatment. AB - Tissues from 50 cases of squamous cell carcinoma of the uterine cervix were analysed for immunohistochemical expression of c-erbB-2 oncoprotein and the patients were followed-up for 2 years. Immunopositivity of c-erbB-2 was studied with reference to clinical stage, histopathological differentiation and response to the cancer therapy. Expression of c-erbB-2 protein was found to be higher (37.5%) in cases with stage II disease, whereas more expressions were noticed in poorly differentiated squamous cell carcinoma (33.3%). Among cases who showed complete response to the treatment, 20.8% were positive for c-erbB-2 oncoprotein. On the contrary, 36.8% of prognostically unfavourable cases revealed positivity for c-erbB-2 immunostaining. However, the difference between c-erbB-2 expressions of these two said groups of patients, which were divided in accordance with the response to treatment, did not attain to statistical significance. Study on c erbB-2 among larger number of patients with cervical carcinoma may prove to be an important factor in response to cancer therapy. PMID- 11480226 TI - Effect of fluvoxamine and N-methyl-D-aspartate receptor antagonists on shock induced depression in mice. AB - We have earlier demonstrated that NMDA receptor antagonists possess antidepressant effect and also they show a synergism with imipramine. The present study attempts to investigate whether NMDA receptor antagonists also interact with selective serotonin reuptake inhibitors. The study was conducted in albino mice using shock-induced depression model. The mice were placed on a grid floor and shock delivered were of 2 sec duration with a 9 sec interval for 1 h. Twenty four hours later depression was measured by an open field test followed by a forced swimming test. Presentation of inescapable foot shock significantly reduced ambulation (from 159.50 +/- 5.42 to 80.50 +/- 4.61) and rearing (from 22.10 +/- 2.15 to 11.30 +/- 1.32) in the open field arena and increased immobility duration in the forced swimming test (from 82.20 +/- 3.51 to 158.90 +/ 4.61). Pretreatment with fluvoxamine, MK-801, ketamine and the combination of fluvoxamine with either of the NMDA antagonists antagonised shock-induced depression. Haloperidol and ketanserin pretreatment modified the effect of these agents. These findings suggest an interaction of NMDA receptor antagonists with fluvoxamine, and an involvement of brain dopaminergic and tryptaminergic mechanisms in the behavioural suppression observed after inescapable foot shock. PMID- 11480227 TI - Short-term REM sleep deprivation effect on temperature rhythm of rats. AB - Circadian rhythm of body temperature (CRT) is altered in endogenous depression and many psychiatric disorders. Even the sleep pattern is disrupted. Sleep deprivation alleviates symptoms in depression. The present study was conducted to find the role of noradrenergic innervation to the pineal gland in bringing about the effect of REM sleep deprivation (REMSD) on the CRT. Adult male Wistar rats (n = 12) divided into 2 groups were used for the study. The group I rats (n = 6) underwent superior cervical ganglionectomy and the group II rats (n = 6) were sham ganglionectomised. After recovery rats were given REMSD for 48 hours. The CRT was measured in three periods of the study i.e. basal, post operative and post REMSD. The results indicated REMSD increased the Amplitude and Mesor of the CRT in both the groups which was shortlasting and reversible thus suggesting non sympathetic mediation of the pineal in bringing about the circadian rhythm alteration due to REMSD. PMID- 11480228 TI - Pulmonary functions in workers of fertiliser and chemical industry. AB - The present study was undertaken to assess the pulmonary functions in fertiliser and chemical industry workers of Kerala. Pulmonary function test was performed with Vitalograph Compact-II spirometer. Exposure to different chemicals, noxious gases and particulate matter of fertiliser and chemical industry was injurious to the normal pulmonary function. Significant decline in lung volumes, viz. FVC, FEV1, FEV1/FVC% (P < 0.01) and flow rates, viz. FEF25-75%, FEF0.2-1.2, FEF25%, FEF50%, FEF75% (P < 0.01) among non-smokers and smokers than that of normal controls was observed. The findings of this study also suggest that with increased duration of exposure to industrial pollutants, lung volumes and flow rates were significantly reduced in workers. PMID- 11480229 TI - Effect of heat stress on gastro-intestinal motility in young albino rats. AB - Motility of the gastrointestinal tract was studied by barium meal technique in control and heat exposed young and adult albino rats. As compared to the controls, the gastric emptying and the intestinal transit were significantly increased in heat exposed young rats, while in vivo intestinal absorption of D glucose and L-proline was significantly decreased in heat exposed young rats. The rapid transit of meal through small intestine in heat exposed young rats could be responsible for decreased absorptive capacity of small intestine by reducing the exposure time for nutrients. PMID- 11480230 TI - Predator induced stress and its analgesic potential on estrous and anestrus albino rats. AB - It has been established that physical stress induces antinociceptive effect. In the present study efforts were made to investigate the role of chronic intermittent psychological stress in the induction of the analgesic effect and the probable role of estrous cycle in modulating the antinociceptive response. Albino rats in regular estrous cycle (n = 15) and those in anestrus (n = 15) were exposed to psychological stress for a period of 20 min each day for 12 consecutive days. The predator (domestic cat) was used to induce the psychological stress. At the end of each session tail flick response time to heat was recorded as a measure of pain perception. It was observed that female estrous rats had a low pain threshold in the beginning but its tolerance increased gradually reaching a peak by 6th to 7th day and returned to control level by 11th day. The anestrus female had higher pain threshold at the beginning and showed a gradual decline to reach the control level as estrous females by 12th day. These results suggest that the ovarian hormones and the corticosterone may modulate the impact of stressor on endogenous pain inhibition and other stress responsive systems. PMID- 11480231 TI - Assessment of physiological stress in periparturient cows and neonatal calves. AB - Pregnancy is considered to be one of the physiological stressors. The stress hormone, cortisol is significantly involved in various events during periparturient period including initiation of parturition. The study was conducted to estimate the serum cortisol concentration in cows and the neonatal calves in order to correlate the effect of cortisol on certain haematological and biochemical parameters such as blood glucose level (BGL), total plasma protein (TPP), lymphocyte:neutrophil ratio and mitogen induced lymphocyte proliferative response. Blood samples were collected from six cows in four periods, namely., 3 days prior to parturition, on the day of parturition, and 7 days after parturition. Blood samples were also collected from neonatal calves in the period 0, 7 and 14 days of age. Calves above two months of age and non-pregnant dry cows were considered as the controls. The serum cortisol concentration in cows on the day of parturition was significantly higher (P < 0.01) than controls and the value in calves was also significantly higher (P < 0.01) at 0 day than their controls. On the day of parturition BGL level of the dam and calves were significantly higher (P < 0.01), whereas the proliferative response of lymphocytes to mitogen was significantly lower (P < 0.01) than controls. However TPP levels did not differ significantly. This confirmed that the dam at the time of parturition and neonatal calf before taking colostrum are under a high risk of infection because of the low profile of immune status. The lymphocyte:neutrophil ratio also justified the above suggestion. PMID- 11480232 TI - Blood lactate concentration at selected of olympic modes weightlifting. AB - This study highlights the blood lactate response of weightlifters (N = 5) in two modes of olympic lifts: Snatch (SN) and Clean & Jerk (CJ), during three types of training namely (1) one repetition lift (ORL), (2) Multiple set session (MSS) and (3) one set session (OSS). In ORL, 30, 40, 50, and 60 kg, each of one repetition only, were lifted with an interval of 5 min between two consecutive loads. Both MSS and OSS consisted of 6 sets of lift: 50% x 6 (i.e. 50% of 1 Repetition Maximum x 6 repetitions), 60% x 5, 70% x 4, 80% x 3, 90% x 2, and 100% x 1. In MSS, 3 to 3.5 min interval was given between two successive sets whereas in OSS the interval was approximately 24 hours. Lactate levels were very low (< 3.5 mM) in ORL. In MSS, lactate reached peak at an intermediate set, but, it was maximum at the first set and then declined gradually in OSS. In most of the cases, however, lactate were significantly higher in CJ than SN. The study concludes that: (a) anaerobic glycolysis is not stimulated considerably when the lifting time is only 4-5 sec, (b) repetition of lift plays more important role, than intensity, in lactate production, (c) CJ is more strenuous than SN for a given %RM. PMID- 11480233 TI - A comparative study of cardiovascular stress during different swimming strokes. AB - The study was aimed: (i) to compare maximum heart rate during exhaustive running (HRmasr) and swimming (HRmaxs), (ii) to compare heart rate (HR) in various modes of swimming-front crawl (FC), butterfly (BF), breast stroke (BRS) and back stroke (BS). HRmaxr (192.3 +/- 4.9 beats/min) of the swimmers (N = 7) was higher, although not significantly (P < 0.05), than their HRmaxs (183.7 +/- 3.8). Peak HR of the swimmers in FC, BF, BRS and BS (each of 100 m distance) were 179.9 +/- 7.2, 180.3 +/- 7, 178.6 +/- 6.2 and 177.3 +/- 5.4 beats/min respectively. Average HR in the respective events were 167.3 +/- 15.8, 166.6 +/- 14.1, 163.3 +/- 13.5 and 165.8 +/- 14.6 beats/min. Differences were not significant between any two modes, either in peak HR or mean HR. The results conclude that: (i) maximum HR in swimming is approximately 8 beats/min lower than maximal running and (ii) when the effort is maximum, cardiovascular stress remains unaltered in different swimming modes. PMID- 11480234 TI - Antimicrobial effect of protein(s) isolated from a marine mollusc Telescopium telescopium. AB - Ammonium sulfate precipitated protein (SF-50) isolated from the spermatheca gland of Telescopium telescopium, an invertebrate marine snail, showed antimicrobial effect on Escherichia coli. The antimicrobial effect varied with the concentration of "SF-50" used and the effect was found to be comparable to antibiotics like amikacin, contrimoxazole and gentamycin in disc diffusion test. The "SF-50" was devoid of erythrocyte haemolysis property. PMID- 11480235 TI - Reversal of paclitaxel induced neutropenia by Withania somnifera in mice. AB - The effect of aqueous extract of Withania somnifera (L. Solanaceae) was studied against paclitaxel induced neutropenia in mice. After paclitaxel 1 mg/kg, i.v. administration significant fall in total WBC and absolute neutrophil count was observed on day 3 and day 5. W. Somnifera (200 mg/kg, p.o.) per se produced significant increase in neutrophil counts. W. somnifera (200 mg/kg, p.o.) when administered for 4 days before paclitaxel treatment and continued for 12 days caused significant reversal of neutropenia of paclitaxel. The findings of the study suggest the potential of W. somnifera as an adjuvant during cancer chemotherapy for the prevention of bone marrow depression associated with anticancer drugs. PMID- 11480236 TI - Dental caries and dietary habits in school going children. PMID- 11480237 TI - [History of the teaching of odontotechnology at the University of Budapest]. AB - The teaching of odontotechnology at the University of Budapest has a more than 100 years history. It was started in the Academic Dental Institute led by Jozsef Arkovy. The first lecturer and practice instructor was Lajos Hattyasy. The first Institute of Odontotechnological Research was established in 1900, and was headed by the first "private docent" of the World appointed in Odontotechnology, Lajos Hattyasy. The Clinic of Stomatology in Budapest was opened in 1909 with European standard dental laboratory and practice rooms. The training program was extended to six semesters according to the act of the 1924 and 1936 Law of Education. In addition to odontotechnology the courses of dental materials and metals as well as restorative dentistry became independent preclinical subjects with a total of 670 teaching hours, making up 17% of the whole dental curriculum. That time Henrik Salamon and Denes Mate were the chief lecturers. After World War II a new type of dental education started in 1952. The curriculum was extended to 10 semester with a great emphasis on odontotechnology. A very modern, phantom head preclinical training laboratory was opened in 1953. The total teaching time of odontotechnology was 360 hours and its theoretical principles were summarized in Hungarian second generation textbooks. That time Imre Foldvari and Gyorgy Huszar were the chief lecturers. From 1965 the significance of the course of odontotechnology was gradually diminishing and the teaching hours were markedly reduced. In 1967 odontotechnology as an independent subject was ceased and integrated into the prosthodontic dentistry. Its teaching time was reduced to 120 hours. In 1996 odontotechnology parallel with other preclinical subjects became independent again and earned a total of 260 teaching hours in the dental curriculum. PMID- 11480238 TI - [Acute effect of calcium hydroxide-containing material in the rat's dental pulp microcirculation]. AB - Experimental stimulation and clinical procedures applied on the crown of the tooth cause vascular reaction in the dental pulp. Laser-Doppler flowmetry is a good method for determining the blood flow of the dental pulp. The aim of these experiments was to study the acute effect of a calcium hydroxide-containing pulp capping material (Dycal, DeTrey) on blood flow of the dental pulp after the application into a deep test-cavity. Two groups of male Sprague-Dawley rats (308 g +/- 50 S.E.) were used: control- and test-group, n = 10. A standardised deep class five cavity was prepared in the left lower incisor of each rat. Laser Doppler flowmeter was used to measure the vascular reaction of the pulp. The levels of blood flow were recorded prior to (0 min.) and after the application (1, 5, 15, 30, 60 min.) of Dycal. Results were evaluated with one-way ANOVA. Significantly higher pulpal blood flow was found only in the first minute after the application of Dycal. In any other time no significant difference was found between the results before and after (5, 15, 30, 60 min.) the application in test and control-group. PMID- 11480239 TI - [Dental implantation at the maxilla with autologous bone transplantation]. AB - In the Department of Maxillofacial and Oral Surgery of County Hospital of Szentes, different types of bone losses of the maxillary alveolar ridge have been treated since 1 March, 1990. The applied surgical procedures were the following: 1. 394 patients were treated with GBR technique and 567 DenTi implants were inserted for these patients. 2. In 47 cases autogenous bone grafts were harvested from mandibular cortica and 78 DenTi implants were placed. Altogether 14 implants were lost; the success rate was 97.81% in the period between 1 March, 1990 and 31 January, 2001. 3. The severely resorbed maxillary alveolar ridge was replaced by rigid autogenous bone graft harvested from the ilium or from tibia. The bone graft was fixed with implants to the recipient surface. This method was applied in 36 cases from 1 February, 1992 until 31 January, 2001. 147 DenTi implants were inserted and 7 of them were lost. The success rate was 97.28%. Our results have indicated that the dental implants had accelerated the remodelling process of the transplanted autogenous bone and the success rate of the simultaneously performed dental implants were comparable to that of the two stage procedures in which dental implants were inserted several weeks following bone grafting. Based on our long-term favourable clinical experience this technique has been proven to be an excellent method for correcting and replacing the lost maxillary alveolar ridges as well as the circumscribed alveolar bony defects. PMID- 11480240 TI - [Endodontic treatment of gangrenous teeth--literature review]. AB - This overview was put together to update the endodontic knowledge of the general practitioners. Several treatment methods sometimes with contradicting principles are applied in Hungary using a wide range of endodontic medicines. The aim of the paper was not only to clarify the basic therapeutic principles but also to provide practical information about the relevant endodontic products for the everyday use. PMID- 11480241 TI - [Traumatic injuries of deciduous teeth]. AB - Review of literature regarding frequency of traumatic injuries of primary teeth, types of injuries, therapy and possible consequences in permanent dentition. The diagram of prevalence shows no direct correlation to age, whereas according to diagram of incidence 70% of injuries occur in the second and third years of life. Injuries without dislocation of the deciduous tooth: fractura coronae dentis decidui, contusio seu concussio dentis decidui, fractura radicis decidui. Injuries with dislocation: luxatio totalis dentis decidui, luxatio partialis dentis decidui, intrusio dentis decidui. Consequences of traumas can be detected in disturbances of second dentition, in developmental anomalies of enamel and in considerable deformities of homologous permanent tooth. PMID- 11480242 TI - [Effect of smoking on the development of oral leukoplakia]. AB - The increase in cancer mortality throughout the world justifies the study of its causes and development. Oropharyngeal cancer is ranked on the sixth place, Hungary with the highest mortality among 46 countries. Tobacco use is implicated in the development of oral cancer, and oral leukoplakia as well. The aim of the study was to give an overview on the connection between tobacco use and oral leukoplakia. Summarizing the literary data strong evidence has been found on the role of smoking in the development both of oral cancer and oral leukoplakia. Epidemiologic patterns of cigarette-smoking show a steep increase in Central European countries, especially in Hungary. Cross-sectional studies indicate higher prevalence rate of leukoplakia among smokers, with a dose-response relationship between tobacco use and oral leukoplakia, while intervention studies show a regression of the lesion after stopping the smoking habit. PMID- 11480243 TI - [Bacteriologic investigation of periodontal infections]. AB - The gingivitis and periodontitis are very common diseases in Hungary: more than 80% of the adult population suffers with gingivitis and 15-20% has destructive periodontitis. The composition of peridontopathogenic bacterial flora is totally different from that of the bacterial flora causing dental infections. In periodontal infections dominantly facultative and obligatory anaerobic microorganisms occur (Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans), Porphyromonas gingivalis (P. gingivalis), Bacteroides forsythus (B. forsythus). The main objective of the study was to investigate the microbial flora of the rapidly progressing periodontitis with special respect to the occurrence of the Actinobacillus actinomycetemcomitans. The antibiotic susceptibility of the A. actinomycetemcomitans was also determined against the most frequently used conventional antibiotics. Twenty-five samples collected from patients with rapidly progressing periodontitis were examined. Prevalence and ratio of A. actinomycetemcomitans and other species of the periodontal anaerobic flora were investigated. MIC values of different antibiotics used routinely- clindamycin, amoxicillin/clavulanic acid, tetracycline, metronidazole--were measured. Prevalence of black pigmented bacteroides (50%), and A. actinomycetemcomitans (30%) was comparable to data of foreign investigators. A. actinomycetemcomitans spp. appear to be more sensitive to clindamycin and amoxicillin/clavulanic acid and more resistant to metronidazole compared to published data. The knowledge of microbial composition of the periodontal flora could help to diagnose the different forms of periodontitis. It can also assist the indication of the most appropriate antibiotic therapy. PMID- 11480244 TI - Comparison of official coders versus physician panel in assignment of underlying cause of death. AB - BACKGROUND AND PURPOSE: In outcome analyses of clinical trials and mortality follow-up studies, the underlying cause of death (UCOD) is commonly assigned either by official coders or by a panel of physicians. We evaluated the validity of UCOD assigned by official coders by comparison with the assignments of a panel of physicians who reviewed the available medical records of the deceased. METHODS: The comparisons focused on deaths occurring from October 1995 through June 1998 in a series of residents in a veterans home. Because of limited resources, only the first 104 deaths that occurred during the study period were included. Agreement rate, sensitivity, specificity, and kappa statistics were calculated to assess the consistency of coder versus physician panel assignment of UCOD by selected main causes of death. For 32 of the 104 deaths, the panel concluded that the information obtained from medical records was insufficient to determine the UCOD, and the following analyses were confined to the other 72 deaths. RESULTS: For the 72 deaths considered by the panel to have sufficient information to determine UCOD, all four physicians agreed on a single UCOD in 50 (69%) cases, while three or four agreed in 66 (92%) cases. A consensus was reached in cases with disagreement. The two procedures completely agreed in 40 (56%) of the deaths. For general category UCOD, the kappa value was high for cancer (0.83) and cardiovascular disease (CVD, 0.73) but only moderate for pulmonary disease (PD, 0.60). When the UCOD assigned by the panel was used as the gold standard, official coders showed relatively low sensitivity for correct determination of UCOD in cases of CVD (0.76) compared with cancer (0.86) and PD (0.80). CONCLUSIONS: Given the high inter-physician consistency and the relatively low sensitivity of official coders in assigning CVD as the UCOD, we conclude that the use of clinical review panels would provide more accurate UCOD assignments for use in outcome analyses in mortality follow-up studies and clinical trials in Taiwan. PMID- 11480245 TI - Sensitivity of diffusion-weighted magnetic resonance imaging in the diagnosis of acute lacunar infarcts. AB - BACKGROUND AND PURPOSE: Heightened interest in the early diagnosis and treatment of acute stroke challenges neuroimaging specialists to optimize available modalities and to develop new techniques for the evaluation of cerebrovascular disease. The purpose of this study was to evaluate the sensitivity of diffusion weighted (DW) magnetic resonance (MR) imaging in detecting early small infarcts and in differentiating acute from nonacute small infarcts when conventional MR imaging demonstrates multiple small infarcts. METHODS: Thirty-eight consecutive patients with a clinical diagnosis of lacunar infarcts (20 men and 18 women, aged 50-79 yr) who underwent DW MR imaging within 3 days of symptom onset were enrolled in this study. All patients underwent both conventional fast spin-echo (FSE) MR imaging and DW MR imaging. Apparent diffusion coefficient (ADC) maps were also acquired. All patients had at least one of the following classic lacunar syndromes: pure motor hemiparesis, ataxic hemiparesis, dysarthria-clumsy hand, pure sensory stroke, and sensorimotor stroke. RESULTS: Thirty-six patients (40 acute lesions) had focal areas of high intensity on DW MR imaging associated with their clinical symptoms. Acute lacunar infarcts were seen on DW MR imaging as bright areas of decreased ADC ratio (range 0.31-0.85, mean 0.64). Lesion conspicuity with DW MR imaging was superior to that with FSE in 33 acute lesions. In four patients with small hyperacute (within 6 hours) infarcts, DW MR imaging was particularly sensitive for infarcts that were not visible on FSE sequences. The sensitivity of DW MR imaging and ADC map for acute lacunar infarcts was 95%, specificity 94%, positive predictive value 97%, negative predictive value 90%, and accuracy 95%. In 15 patients with both acute and nonacute old small infarcts, DW MR imaging and ADC map could easily distinguish the new infarct from adjacent old ones, although this distinction was difficult to make with FSE. CONCLUSIONS: DW MR imaging accompanied by ADC map is a sensitive diagnostic modality for hyperacute and acute lacunar infarcts. It is also sensitive in distinguishing fresh small infarcts from adjacent multiple old infarcts. PMID- 11480246 TI - Influence of cytokines on matrix metalloproteinases produced by fibroblasts cultured in monolayer and collagen gels. AB - BACKGROUND: Extracellular matrix metalloproteinases (MMPs) are crucial factors involved in connective tissue remodeling that accompanies ultraviolet radiation induced actinic damage. This study investigated whether the cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, and IL-10 modulate the expression of MMPs in cultured human newborn skin fibroblasts. METHODS: Different concentrations of TNF-alpha, IL-1 beta, and IL-10 were added to human dermal fibroblasts grown in monolayers or embedded in three-dimensional (3D) collagen gels, a model closer to skin. Supernatant from the fibroblast cell culture was collected 24 hours later. The concentrations of MMP-1 and MMP-3 were assaysed by enzyme-linked immunosorbent assay (ELISA) while the concentrations of MMP-2 and MMP-9 were analysed by zymography. RESULTS: Basal production of MMPs was significantly greater in fibroblasts grown in 3D gels than in cells grown as monolayers. TNF-alpha and IL-1 beta induced increases in the concentrations of MMP-1, MMP-3, and MMP-9, but not in MMP-2 or tissue inhibitor of matrix metalloproteinase (TIMP)-1 or -2. The inducibility of MMP secretion is more significant in 3D gels. IL-10 did not significantly modulate MMPs. CONCLUSION: This study demonstrated that basal concentrations of MMPs are higher in fibroblasts cultured in 3D gels and their response to cytokines is different to that of cells grown as monolayers. Cytokines can increase the collagenolytic and gelatinolytic activity involved in extracellular matrix remodeling and hence contribute to photoaging. PMID- 11480247 TI - Cytotoxic and antioxidant effects of the water extract of the traditional Chinese herb gusuibu (Drynaria fortunei) on rat osteoblasts. AB - BACKGROUND AND PURPOSE: Gusuibu (Drynaria fortunei) is a traditional Chinese herb that has been claimed to have therapeutic effects on bone healing; however, a clinical mechanism responsible for this effect has not been identified. This study evaluated the cytotoxic and antioxidant effects of the water extract of gusuibu (WEGSB) on rat osteoblasts. MATERIALS AND METHODS: Osteoblasts were prepared from neonatal Wistar rat calvarias and treated with WEGSB. Cell viability and alkaline phosphatase activity were determined. Intracellular reactive oxygen species were detected using the dye 2',7'-dichlorofluorescin, and mitochondrial membrane potential was detected using the dye 3,3' dihexyloxacarbocyanine iodide and flow cytometry. RESULTS: WEGSB at 1 and 10 micrograms/mL was not cytotoxic to rat osteoblasts, but WEGSB at 100 micrograms/mL reduced cell viability and alkaline phosphatase activity in a time dependent manner. Although WEGSB and hydrogen peroxide did not affect the mitochondrial membrane potential of rat osteoblasts, combined treatment with WEGSB (100 micrograms/mL) and hydrogen peroxide lowered the membrane potential of mitochondria and resulted in cell death. The basal level of intracellular reactive oxygen species in rat osteoblasts was significantly suppressed by WEGSB at 10 to 100 micrograms/mL. WEGSB (10 micrograms/mL) specifically inhibited hydrogen peroxide-induced oxidative stress without an effect on nitric oxide induced stress. Hydrogen peroxide caused concentration-dependent death of rat osteoblasts, but WEGSB significantly protected cells from hydrogen peroxide induced death. CONCLUSION: This study has shown that WEGSB at 10 micrograms/mL is not cytotoxic to rat osteoblasts in vitro, and also that the extract at 10 micrograms/microL has an antioxidant effect on these cells. The antioxidant activity of WEGSB can protect rat osteoblasts from hydrogen peroxide-induced death and may promote bone recovery under similar pathologic conditions. PMID- 11480248 TI - Surgical treatment of spinal arteriovenous malformations: vascular anatomy and surgical outcome. AB - BACKGROUND AND PURPOSE: Spinal arteriovenous malformations (AVMs) are rare but debilitating lesions of the central nervous system. This study evaluated the outcome in patients with spinal AVMs treated surgically, and the relationship between outcome and the vascular anatomy of the lesions. METHODS: These AVMs were classified into four types: dural AVM (type I), glomus AVM (type II), juvenile AVM (type III), and intradural direct arteriovenous fistula (type IV). Either interruption of the feeding vessels or excision of the AVMs was performed in all patients. RESULTS: Intradural AVMs manifested as subarachnoid or intramedullary hemorrhages, whereas dural AVM manifested as epidural hemorrhage in two patients, and as an episode of subarachnoid hemorrhage in one patient. The nidus in five of the six dural AVMs was below the mid-thoracic level. In six of the seven patients with intradural AVMs, the nidus was located in the cervical region. The prognosis of patients with dural AVMs was generally good, but in patients with intradural AVMs, motor recovery was worse and resection was more difficult. CONCLUSIONS: Correct diagnosis and classification of spinal AVMs are the key prerequisites for successful treatment. The surgical outcome is closely related to the vascular anatomy of the lesion. For dural AVMs, only surgical interruption of the arteriovenous shunting at the dural sleeve of the nerve is required and good surgical results are often obtained. Intradural direct arteriovenous fistula responded well to surgery, whereas combined endovascular and microsurgical techniques can minimize the chance of rebleeding in intramedullary AVMs. PMID- 11480249 TI - One-stage correction of neglected developmental dysplasia of the hip by open reduction and pemberton osteotomy. AB - BACKGROUND AND PURPOSE: Pemberton osteotomy is commonly used for correction of hip-dysplasia. Its application in treating hip dislocation is rarely reported. The purpose of this study was to determine the efficacy of one-stage open reduction and Pemberton osteotomy of neglected developmental dysplasia of the hip (DDH). METHODS: We retrospectively reviewed the clinical and radiographic results of one-stage total correction in 75 patients with neglected DDH (86 hips) from 1984 through 1996. Sixty-nine patients (77 hips) aged 13 to 76 months (mean, 21.5 mo) underwent open reduction, Pemberton osteotomy, and hip spica for 8 weeks. Seven patients (9 hips) aged 29 to 73 months (mean, 46.6 mo) underwent additional femoral shortening (another hip in one bilaterally involved patient underwent only open reduction and Pemberton osteotomy). RESULTS: Acetabular index improved markedly from 40.4 degrees (range, 25-60 degrees) preoperatively to 16.2 degrees (range, 0-30 degrees) postoperatively. Complications included redislocation in four patients (4 hips) and avascular necrosis in three patients (3 hips). Reoperation was performed in four patients (4 hips). All patients were followed up for at least 3 years. By Severin's classification, all operated hips had excellent (18) or good (68) final radiographic results. The functional results, evaluated by McKay's criteria, were also excellent (52 patients/60 hips) and good (23 patients/26 hips). CONCLUSIONS: Neglected DDH was safely and effectively treated by open reduction, Pemberton osteotomy, and optional femoral shortening with limited complications. PMID- 11480250 TI - Successful liver transplantation in a child with biliary atresia and hepatopulmonary syndrome. AB - Hepatopulmonary syndrome (HPS) is characterized by hypoxemia in patients with severe chronic liver disease and pulmonary vasodilatation in the absence of primary cardiac or pulmonary disease. Severe hypoxemia resulting from HPS is generally considered a contraindication to liver transplantation. We describe the case of a 6-year-old girl with biliary atresia complicated with HPS who was successfully treated with liver transplantation. Cyanosis and dyspnea had initially developed at the age of 5 years. Arterial blood gas showed a PaO2 of 46.6 mm Hg on room air. The diagnosis of HPS was confirmed by contrast echocardiography, lung perfusion scan with 99mTc macroaggregate albumin, and pulmonary angiography. The lung scan revealed an intrapulmonary shunt of 24%. She underwent living donor liver transplantation and received a left lateral segment graft from her mother. One year after successful liver transplantation, she had normal arterial oxygen saturation and a normal lung scan without intrapulmonary shunting. This case demonstrates that HPS associated with end-stage liver disease is potentially curable by liver transplantation. PMID- 11480251 TI - Mutation analysis in the family of a Taiwanese boy with with epidermolysis bullosa simplex dowling-meara. AB - Epidermolysis bullosa simplex (EBS) is a group of hereditary bullous diseases characterized by intraepidermal blistering due to mechanical stress-induced degeneration of basal keratinocytes. The major subtypes of EBS, including EBS Dowling-Meara (EBS-DM), are caused by mutations of the basal keratin genes, keratin 5 (KRT5) or keratin 14 (KRT14). Here, we describe the first reported pedigree of EBS-DM in Taiwan. The proband was a 5-day-old newborn, who presented with numerous blisters of various sizes, some of which were hemorrhagic, as well as erosions on the extremities and hard palate since birth. Biopsy of a new vesicle showed subepidermal and basal cleavage with infiltration of eosinophils and neutrophils. Electron microscopy revealed cytolysis of basal cells and clumping of tonofilaments forming thick bundles and peculiar electron-dense round or oval basket-weave bodies. These features are characteristic of EBS-DM. The proband's mother had also suffered from a similar blistering disorder since birth, with gradual appearance of mottled pigmentation on the trunk, diffuse irregular or linear palmoplantar hyperkeratosis, and nail dystrophy. Mutation analysis revealed a heterozygous point mutation (R125C) in helix 1A of keratin 14 in the proband and his mother. The detection of this pathogenic point mutation enables future prenatal diagnosis in this family. PMID- 11480252 TI - Lipoma arborescens of the knee: a case report. AB - Lipoma arborescens is a rare intraarticular lesion of unknown etiology. We describe the radiographic, computerized tomographic, and magnetic resonance (MR) imaging characteristics of lipoma arborescens in a 17-year-old boy presenting with chronic painful swollen knee. Histologic examination after surgical synovectomy confirmed the diagnosis of lipoma arborescens. The fatty nature and frond-like appearance of this lesion can be demonstrated on imaging studies, especially MR imaging. Open synovectomy is curative in most cases. Although lipoma arborescens is rare, it is important to recognize and differentiate it from other synovial lesions by MR imaging. PMID- 11480253 TI - Bethlem myopathy in a Taiwanese family. AB - We report three cases of Bethlem myopathy from three consecutive generations of a Taiwanese family, including one woman aged 70, one man aged 40, and a boy aged 8. The clinical features of the patients included autosomal dominant inheritance, childhood or adolescent onset, mainly proximal and extensor involvement, early diffuse joint contractures, and absence of cardiac involvement. These features fulfilled the diagnostic criteria for Bethlem myopathy. Though the clinical course of the disease was once thought to be benign, our female patient became wheelchair-bound at the age of 53. This suggests that the disease process in Bethlem myopathy is slow but ongoing. PMID- 11480254 TI - Endothelin-A receptor antagonist prevents neonatal pulmonary hypertension in meconium aspiration in piglets. AB - In order to test the hypothesis that endothelin-1 (ET-1) directly contributes to the pathophysiology of pulmonary hypertension induced by meconium aspiration, we randomized 12 anesthetized and paralyzed piglets to receive BQ-123, a selective endothelin-A receptor antagonist (BQ group), or normal saline (NS group) after meconium aspiration. The animals were instilled with meconium mixture (3 mL/kg) via an endotracheal tube, and then given intravenous BQ-123 (2 mg/hr) or normal saline. Plasma ET-1 concentrations, arterial blood gases, and hemodynamics were measured at baseline and at 60, 120, 180, and 240 minutes after instillation. The results showed that plasma ET-1 concentrations were similar in both groups. However, in the BQ group, pulmonary artery pressure was significantly lower after 120 minutes (p < 0.05 at 120 min, p < 0.01 at 180 and 240 min) and pulmonary vascular resistance was significantly lower after 180 minutes (p < 0.01) than in the NS group. No significant difference was found in systemic hemodynamics. These data suggest that ET-1 directly contributes to the pathophysiology of pulmonary hypertension induced by meconium aspiration. PMID- 11480255 TI - Anaphylaxis after propofol infusion for Port-A-Cath insertion in a 35-year old man. AB - Anaphylaxis to propofol is rare and has not been previously reported in Asia. We describe a 35-year-old man with nasopharyngeal carcinoma who developed acute respiratory distress and hypotension after propofol infusion for parenteral anesthesia for Port-A-Cath insertion. Chest roentgenogram showed bilateral diffuse alveolar infiltrates. Respiratory failure ensued, and vocal cord swelling was found during endotracheal intubation. Hemodynamic data included a low cardiac index, a low systemic vascular resistance, and a high pulmonary vascular resistance. His condition and the shadows on the chest roentgenogram improved quickly after fluid challenge and the use of vasopressors, antihistamine, and intravenous steroids. Early awareness and appropriate management are necessary to prevent a fatal outcome in patients with propofol anaphylaxis. PMID- 11480256 TI - Hormone-related headache. AB - The normal female life cycle is associated with hormonal milestones including menarche, pregnancy, OC use, menopause, and the use of HRT. Menarche marks the onset of menses and cyclic changes in hormone levels. Pregnancy is associated with rising noncyclic levels of sex hormones. Menopause is associated with declining noncyclic levels. Hormonal OC use during the reproductive years and HRT in menopause are therapeutic hormonal interventions that alter the levels and cycling of sex hormones. These events and interventions may cause a change in the prevalence or intensity of headache. Headaches associated with OC use or menopausal HRT may be related, in part, to periodic discontinuation of oral sex hormone preparations. The treatment of migraine associated with changes in sex hormone levels is difficult, and patients often are refractory to therapy. Based on what is known of the pathophysiology of migraine, a logical approach to the treatment of headaches that are associated with menses, menopause, and OCs using abortive and preventive medications and hormonal manipulations has been presented. PMID- 11480257 TI - Pediatric headache. AB - Headaches are frequent in children and adolescents and at times can be extremely disabling. Disability scales, such as the MIDAS scale, have been useful in helping follow adult patients. Modifications of this scale have been helpful in following pediatric and adolescent patients. Greater attention has been paid to epidemiology and classification of headache in children. Studies are being done on serotonin 1B/1D agonist for treating acute migraine, and this agent has been found to be efficacious despite a high placebo response. It is anticipated that FDA approval of sumatriptan nasal spray in adolescents is forthcoming. Despite advancements, there is no wonder drug. There continues to be a need for studying preventive therapies in a double-blind, placebo-controlled environment, and plans are under way for such studies. Many adult patients with chronic daily headaches report that their headaches began in childhood and adolescence. A better understanding of diagnostic criteria, early diagnosis, and more effective treatment may be the key to influencing the prevalence of headaches in adults. Continued research is the only answer to the questions raised by the most recent studies in this population. PMID- 11480258 TI - Behavioral and nonpharmacologic treatments of headache. AB - Cognitive-behavioral analysis and the multiaxial assessment of relevant behavioral domains (headache frequency and severity, analgesic and abortive use and misuse, behavioral and stress-related risk factors, comorbid psychiatric disorders, and degree of overall functional impairment) help set the stage for CBT of headache disorders. Controlled studies of CBTs for migraine, such as biofeedback and relaxation therapy, have a prophylactic efficacy of about 50%, roughly equivalent to propranolol. Cluster headache responds poorly to behavioral treatment. The persistent overuse of symptomatic medication impedes the effectiveness of behavioral and prophylactic medical therapies. Behavioral treatment can help sustain improvement after analgesic withdrawal, however, and prevent relapse in cases of analgesic overuse. Cognitive factors (e.g., an enhanced sense of self-efficacy and internal locus of control) appear to be important mediators of successful behavioral treatment. Patients with CDH are more likely to overuse symptomatic medication (and in some cases abuse analgesics), have more psychiatric comorbidity; have more functional impairment and disability, and are at least as likely to experience stress-related intensification of headache as patients whose episodic headaches occur less than 15 days per month. Despite the significance of these behavioral factors, patients with CDH (particularly those with migrainous features) are less likely to benefit from behavioral treatment without concomitant prophylactic medication than is the case for episodic TTH and migraine sufferers. Continuous daily pain may be more refractory to behavioral treatment as a solo modality than CDH marked by at least some pain-free days or periods of time. The combination of behavioral therapies with prophylactic medication creates a synergistic effect, increasing efficacy beyond either type of treatment alone. Compliance-enhancement techniques, including behavioral contracts for patients with severe personality disorders, can increase adherence to behavioral recommendations. CBT has earned an important place in the comprehensive treatment of patients with episodic migraine/TTH and severe, treatment-resistant chronic daily headache. PMID- 11480259 TI - Alternative therapies in headache. Is there a role? AB - Alternative or complementary modes of treatment often lack scientific proof of efficacy. This is true for many drugs that are used for prophylaxis of headaches as well. Many of these complementary modes are inexpensive, harmless, and possibly effective. Patients can be given a list of options in the following order of clinician preference: aerobic exercise; isometric neck exercise; biofeedback; a combination product containing magnesium, riboflavin, and feverfew; and acupuncture. The patient must decide which of these approaches is appealing, affordable, and realistically doable. PMID- 11480260 TI - A spectrum of exertional headaches. AB - Headaches that have an explosive onset with exercise, including sexual activity, generally are benign in origin. A subarachnoid hemorrhage, a mass lesion in the brain, or an anomaly of the posterior fossa must be considered, however. The mechanisms that produce sexually induced or cough headaches of abrupt onset are unknown. It is known, however, that a rapid increase in intrathoracic pressure suddenly reduces right atrial pressure and presumably decreases venous sinus drainage from the brain. This situation results in a transient increase in intracranial pressure. Jaw pain that occurs with chewing often is considered to be TMJ dysfunction when arthritic in quality and if subluxations of the jaw can be shown on the physical examination. Giant cell arteritis and common or external carotid artery occlusive disease should be considered when the pain is ischemic in quality. An anginal equivalent is another possibility. Headaches that worsen with vigorous exercise are commonly migrainous. When their onset is apoplectic with exertion (particularly exertion against a closed glottis), the most likely diagnoses are increased intracranial pressure, a posterior fossa abnormality, or benign exertional headaches. Most cardiac induced headaches, but not all, are of a more gradual onset. If there are significant risk factors for coronary artery disease, an exercise stress test is appropriate. A therapeutic trial of nitroglycerin may help to establish a diagnosis if it improves the headache. Using antimigraine drugs as a diagnostic test is inappropriate because triptans and ergots are contraindicated in the presence of coronary artery disease, and a positive response is not diagnostic of migraine. PMID- 11480261 TI - Clinical evaluation of a patient presenting with headache. AB - Evaluation of the headache patient requires a reasonable history and physical examination supported by some knowledge of common headache disorders and their presentations. The diagnostician requires the ability to recognize serious headache disorders based on a working knowledge of the principles of clinical neurology. This article outlines the salient points to consider in the process of headache assessment, based on a clinical overview analysis of the characteristics of several headache disorders. PMID- 11480262 TI - Diagnostic testing for headache. AB - A careful and complete headache history supplemented by a neurologic and general physical examination, as appropriate, enables the astute physician to diagnose most headaches correctly without diagnostic testing. When indications are present (see Box 1), some headache patients with a normal physical examination require testing even though the yield may be low. Failure to test may result in misdiagnosis of potentially serious and life-threatening causes of headaches, such as brain tumors, chronic meningitis, SAH, and temporal arteritis. PMID- 11480263 TI - Epidemiology and impact of primary headache disorders. AB - Migraine is a common disorder that remains largely undiagnosed and has a profound economic impact. In the future, better recognition and treatment can decrease costs and improve the lives of millions of Americans. PMID- 11480264 TI - The clinical spectrum of migraine. AB - The variable clinical features of migraine and some relationships of migraine with other headache disorders have been reviewed. Although these views generally are shared by most headache specialists, some respected headache investigators reject the concept of a spectrum of migraine and conclude that the continuum model is an artifact. PMID- 11480265 TI - Toward a rational understanding of migraine trigger factors. AB - The typical migraine patient is exposed to a myriad of migraine triggers on a daily basis. These triggers potentially can act at various sites within the cerebral vasculature and the central nervous system to promote the development of migraine headache. The challenge to the physician is in the identification and avoidance of migraine trigger factors within patients suffering from migraine headache. Only through a rational approach to migraine trigger factors can physicians develop an appropriate treatment strategy for migraine patients. PMID- 11480266 TI - Mechanism of migraine and action of antimigraine medications. AB - Dilation and inflammation of cephalic arteries and intracranial extra cerebral arteries cause the migraine headache. The migraine-associated symptoms result from the activation of the sympathetic nervous system caused by the pain. The migraine aura is caused by the neurophysiological phenomenon of spreading excitation/depression. The various mechanisms are tied together in the so-called "parallel theory" of the pathogenesis of the migraine attack. The abortive antimigraine medications act by constricting the dilated arteries through stimulation of the serotonin 1B receptor. The preventive antimigraine medications act by increasing the tone of the extacranial arteries, inhibiting the mechanism of neurogenic inflammation, or inhibiting pain transmission within the central nervous system. PMID- 11480267 TI - Safety and rational use of the triptans. AB - The safety of the triptans has been established, with more than 8 million patients treating greater than 340 million attacks with sumatriptan alone. All triptans narrow coronary arteries by 10% to 20% at clinical doses and should not be administered to patients with coronary or cerebrovascular disease. Some triptans have the potential for significant drug-drug interactions (sumatriptan, rizatriptan, and zomitriptan and monoamine oxidase inhibitors; rizatriptan and propanolol; zolmitriptan and cimetidine; and eletriptan and CYP3A4 metabolized medications and p-glycoprotein pump inhibitors). Rational use of triptans should be governed by the use of these medications for patients with disability associated with migraine. Patients with greater than 10 days of at least 50% disability during 3 months have benefited from treating with triptans as their first-line treatment for acute attacks. When the decision has been made to treat with a triptan, the patient should be instructed to treat early in the attack, when the pain is at a mild phase. This approach increases the likelihood of achieving a pain-free response, with fewer adverse events and lower likelihood of the headache recurring. PMID- 11480268 TI - Evaluation and management of headache in the emergency department. AB - In the ED, correct diagnosis is the necessary foundation on which specific therapy is based. There is no substitute for obtaining a thorough history and examining the patient competently. Patients with a past history of primary headaches, such as migraine, also may be afflicted with (new) secondary headaches. Although efficiency is desirable from the perspective of ED management, a thoughtful approach complemented by the judicious selection of tests is compatible with that goal as well as achieving the desired outcome of accurate diagnosis and relief of pain. Arrangements for long-term follow-up are important. PMID- 11480269 TI - Posttraumatic headache. AB - Chronic posttraumatic headache is a common condition, often part of the postconcussion/posttraumatic syndrome. The pathophysiology is not well understood but includes biological, psychological, and social factors. Tension-type headache is the most common manifestation, but exacerbations of migraine-like headaches often occur. After a structural lesion has been ruled out, the treatment of posttraumatic headache is similar to that of the primary headaches. PMID- 11480270 TI - Cluster and related headaches. AB - The disorders described in this article are relatively rare, but probably are more common than previously thought. Because these disorders cause significant pain and disability and treatment response differs from that of migraine, tension type, and cluster headaches, recognition is essential. Table 1 lists the important clinical features of these syndromes and contrasts them with cluster headache, the disorder for which they are often confused. PMID- 11480271 TI - Synchronization and assembly formation in the visual cortex. PMID- 11480272 TI - Divergence and reconvergence: multielectrode analysis of feedforward connections in the visual system. PMID- 11480273 TI - Comparative population analysis of cortical representations in parametric spaces of visual field and skin: a unifying role for nonlinear interactions as a basis for active information processing across modalities. PMID- 11480274 TI - Coordinate transformations in the visual system: how to generate gain fields and what to compute with them. PMID- 11480275 TI - Exploring olfactory population coding using an artificial olfactory system. PMID- 11480276 TI - Neural population coding in the auditory system. PMID- 11480277 TI - Population coding in the auditory cortex. PMID- 11480278 TI - Representations based on neuronal interactions in motor cortex. PMID- 11480279 TI - Connectionist contributions to population coding in the motor cortex. PMID- 11480280 TI - Distributed processing in the motor system: spinal cord perspective. AB - Recordings of spinal INs during a flexion/extension wrist task with an instructed delay period have shown directly that many spinal neurons modulate their rate during the preparatory period soon after a visual cue. The onset time and the relation between the delay period activity of spinal INs and the ensuing movement response suggest that this type of activity is not simply related to the forthcoming motor action, but rather reflects a correct match between the visual cue and the motor response. The existence of such activity further supports the notion that the motor system operates in a parallel mode of processing, so that even during early stages of motor processing multiple centers are activated regardless of their anatomical distance from muscles. The firing properties of spinal INs during the performance of the task seem to differ from the comparable properties of motor cortical cells. Spinal INs fire in a highly regular manner- their CV is substantially lower than the observed CV of cortical cells. Also, although neighboring cells tend to have similar response properties, the frequency of significant correlation is lower than for cortical cells and the anatomical extent of the correlation seems to be narrower. The similarity and differences between cortical and spinal cells in terms of response and firing properties suggests that while both type of cells are active in parallel throughout the behavioral phases of the motor task, each may operate in a different mode of information processing. PMID- 11480281 TI - Coding in the granular layer of the cerebellum. PMID- 11480282 TI - The cerebellum as a neuronal prosthesis machine. PMID- 11480283 TI - Population coding: a historical sketch. PMID- 11480284 TI - Do sleeping birds sing? Population coding and learning in the bird song system. PMID- 11480285 TI - Overcoming the limitations of correlation analysis for many simultaneously processed neural structures. PMID- 11480286 TI - Accuracy and learning in neuronal populations. PMID- 11480287 TI - What ensemble recordings reveal about functional hippocampal cell encoding. PMID- 11480288 TI - Distributed processing in cultured neuronal networks. PMID- 11480289 TI - Long-term chronic multichannel recordings from sensorimotor cortex and thalamus of primates. PMID- 11480290 TI - Temporal and spatial coding in the rat vibrissal system. PMID- 11480291 TI - The evolution and implications of population and modular neural coding ideas. PMID- 11480292 TI - Thalamocortical and corticocortical interactions in the somatosensory system. PMID- 11480293 TI - Reintroducing otolaryngologists to esophageal disease. PMID- 11480294 TI - Temporal bone fracture with displacement of bone. PMID- 11480295 TI - Endoscopic view of an anterior middle turbinate polyp. PMID- 11480296 TI - Acute laryngitis in a professional singer. PMID- 11480297 TI - Normal transnasal esophagoscopy. PMID- 11480298 TI - Cochlear and fenestral otosclerosis. PMID- 11480299 TI - A woman with both positional and spontaneous dizziness. PMID- 11480300 TI - Use of the malleus handle as a landmark for localizing the round window membrane. AB - Localization of the round window membrane is desirable in order to allow a more direct application of medication to the inner ear. A new procedure has been developed to deliver direct, near-continuous inner ear perfusion to the round window with the Silverstein MicroWick. In this office procedure, the wick is inserted through a tympanostomy tube into the round window niche. Accurate localization of the round window is a necessary component of this procedure. In an effort to ascertain the precise location of the round window, we examined 25 cadaveric human temporal bones and measured the distance from the umbo to the round window in each sample. We found that the round window was an average of 3.44 mm (+/- 0.68) from the umbo and was situated at an average angle of 113.2 degrees (+/- 9.8) from the long process of the malleus. Our simple and reliable determination of the relationship between the malleus handle and the round window niche allows for the accurate placement of the Silverstein MicroWick and other devices. PMID- 11480301 TI - Superficial leiomyosarcoma of the head and neck: case report and review of the literature. AB - Superficial leiomyosarcomas are rare in the head and neck region. Because of the infrequent nature of soft tissue sarcomas in general, superficial leiomyosarcomas are often misdiagnosed on clinical grounds. Immunohistochemistry is essential for an accurate histologic diagnosis, and it should include a broad panel of antibody studies. With respect to differences in clinical appearance and biologic behavior, superficial leiomyosarcomas can be broadly classified as either cutaneous or subcutaneous; local control and overall survival are significantly more favorable in patients with the former. The primary treatment of a leiomyosarcoma is a wide surgical excision with an emphasis on negative margins. Treatment failures are usually attributable to a local recurrence. Systemic metastasis occurs in about one-third of patients with subcutaneous involvement. Although cutaneous leiomyosarcoma is considered a relatively more benign process with minimal metastatic potential, systemic metastasis is still possible. This was demonstrated in our case, as a recurrent cutaneous leiomyosarcoma metastasized to the lung. Proper management requires inclusion of this entity in the differential diagnosis, as well as familiarity with its clinical behavior. In this article, we review the literature on superficial leiomyosarcoma and discuss its epidemiology, presentation, clinical behavior, evaluation, tissue diagnosis, staging, and treatment. PMID- 11480302 TI - Granular cell tumor of the parotid: a case report and literature review. AB - Granular cell tumors are relatively uncommon soft tissue lesions that originate in Schwann's cells. Although these tumors can occur at any site, their presence in the parotid gland is very unusual. This article describes one such case in a 48-year-old woman, followed by a brief review of the literature on this subject. PMID- 11480303 TI - Nasal T-cell lymphoma: case report and review of diagnostic features. AB - A 73-year-old man was referred to us for evaluation of extensive nasal crusting and progressive erosion of the nasal midline structures. Clinical examination suggested that the patient had a T-cell lymphoma, a suspicion that was confirmed on immunohistochemical analysis. The patient was treated with combination chemo- and radiotherapy and exhibited a marked response. At the 14-month followup, he remained disease-free. PMID- 11480304 TI - A rare case of renal cell carcinoma metastatic to the sinonasal area. AB - We report a rare case of a hypernephroma that had metastasized to the sinonasal area as a solitary mass. Despite much difficulty, a confirmatory biopsy of the nasal mass was obtained. After radiotherapy had failed to arrest the tumor's growth, the mass was removed via a craniofacial resection. This case illustrates the importance of (1) a high index of suspicion in differentiating this rare tumor from other benign or malignant diseases, (2) awareness of the risk of hemorrhage and the role of preoperative embolization, (3) the need for paraffin histology rather than fine-needle aspiration or frozen section diagnosis, (4) individualizing therapy after due consideration of the characteristics of both the patient and the tumor, and (5) keeping abreast of the most current therapeutic modalities so that patient management can be individualized and optimized. Surgical excision offers the best hope for long-term survival, and it reduces the degree of pain, epistaxis, and disfigurement from the expanding tumor. When such tumors are not resectable, management with radiotherapy, hormonal therapy, immunotherapy, or chemotherapy can be considered. PMID- 11480305 TI - A neurilemmoma of the parotid gland: report of a case. AB - Benign primary tumors of the facial nerve are frequently misdiagnosed because of the variety of their clinical manifestations. Much attention has been paid to neurilemmomas in the recent otolaryngologic literature, but far less has been focused on intraparotid facial nerve neurilemmomas. In this article, we describe a new case of this truly rare tumor, and we review its diagnosis, pathology, and treatment. PMID- 11480306 TI - Benchmarking to the world's best in mathematics. Quality control in curriculum and instruction among the top performers in the TIMSS. AB - This article describes the education quality control systems (for mathematics) used by those countries that performed best on the Third International Mathematics and Science Study (TIMSS). Enforced quality control measures are defined as "decision points"--where adherence to the curriculum and instruction system can be reinforced. Most decision points involve stakes for the student, teacher, or school. They involve potential consequences for failure to adhere to the system and to follow the program at a reasonable pace. Generally, countries with more decision points perform better on the TIMSS. When the number of decision points and TIMSS test scores are adjusted for country wealth, the relationship between the degree of (enforced) quality control and student achievement appears to be positive and exponential. CONCLUSION: The more (enforced) quality control measures employed in an education system, the greater is students' academic achievement. PMID- 11480307 TI - An evaluation of analysis options for the one-group-per-condition design. Can any of the alternatives overcome the problems inherent in this design? AB - This article addresses the analytic problems associated with a design in which one identifiable group is allocated to each treatment condition and members of those groups are measured to assess the intervention. Such designs are often called quasi-experiments if the groups are not randomized to conditions and group randomized trials if the groups are randomized. They present special problems, and previous reports have argued against their use in efficacy or effectiveness trials. Even so, this design still appears with surprising frequency. This article presents the results from a new simulation study that underscores the analytic problems associated with this design. PMID- 11480308 TI - The effect of data collection mode on smoking attitudes and behavior in young African American and Latina women. Face-to-face interview versus self administered questionnaires. AB - Evaluating smoking prevention and cessation programs requires valid data collection. This study examined two survey modes--face-to-face (FTF) interview and self-administered questionnaire (SAQ)--comparing response rates, sample characteristics, data quality, and response effects. From two family planning clinics, 601 female Latina and African American clients ages 12 to 21 were recruited and randomized to either group. Results reveal that neither mode is superior to the other. The SAQ may therefore be preferable for this population, despite its higher rate of incompletes, because it yields results similar to the FTF yet is more cost effective and less disruptive to clinic routines. PMID- 11480309 TI - Probabilistic cost-effectiveness analysis of HIV prevention. Comparing a Bayesian approach with traditional deterministic sensitivity analysis. AB - In cost-effectiveness analysis, the incremental cost-effectiveness ratio is used to measure economic efficiency of a new intervention, relative to an existing one. However, costs and effects are seldom known with certainty. Uncertainty arises from two main sources: uncertainty regarding correct values of intervention-related parameters and uncertainty associated with sampling variation. Recently, attention has focused on Bayesian techniques for quantifying uncertainty. We computed the Bayesian-based 95% credible interval estimates of the incremental cost-effectiveness ratio of several related HIV prevention interventions and compared these results with univariate sensitivity analyses. The conclusions were comparable, even though the probabilistic technique provided additional information. PMID- 11480310 TI - Viable M. leprae as a research reagent. AB - Mycobacterium leprae remain a rare research resource. They cannot be cultivated on artificial media, and the only established means to quantify viability of M. leprae has been by its relative growth in the foot pads of conventional mice (MFP). The MFP method is technically difficult and requires several months to yield results. More effective methods are needed. We examined the association between M. leprae's ability to oxidize 14C-palmitate in axenic culture and the MFP growth results of a large number of suspensions. Oxidative activity was assessed by radiorespirometry (RR) using the Buddemeyer-type biphasic culture vessels containing 7H12 liquid medium and 14C-palmitate, or with commercially prepared BACTEC 12B vessels containing the same medium. The RR results were highly correlated (r = 0.71) with the growth level that each M. leprae suspension achieved by the MFP technique. In using this technique to examine the effects that many common laboratory practices have on M. leprae viability, we found that viability varies markedly between bacillary suspensions derived from different hosts and tissues. The highest viabilities were obtained with bacilli from moderately enlarged nude MFP (< 1 g). Viability tended to be lower among very large nude MFP or long-duration infections and from armadillo tissues. After their harvest from host tissues, leprosy bacilli lost viability quickly. Suspensions stored in 7H12 liquid medium retained < 1% of their viability within 3 weeks of harvest, and freezing bacillary preparations or incubating them at 37 degrees C resulted in nearly an immediate equivalent loss in metabolic activity and viability. M. leprae viability is maintained best when bacilli are stored for only short periods of time at 4 degrees C-33 degrees C. Palmitate oxidation is a rapid, reliable and objective means by which to estimate the viability of M. leprae and can be used effectively as a surrogate for the conventional MFP technique in many studies. PMID- 11480311 TI - Limited ATP generation in cells of Mycobacterium leprae Thai-53 strain in enriched Kirchner liquid medium containing adenosine. AB - The ATP generation in cells of Mycobacterium leprae Thai-53 strain takes place in vitro when the cells are cultivated in Kirchner liquid medium, pH 7.0, enriched with egg-yolk solution, pyruvate, transferrin, and adenosine at 30 degrees C. Among the supplements, adenosine was key and critical for the ATP generation. The optimal concentration of adenosine was 50 micrograms/ml of the medium. ATP generation, however, was limited; the rates of increase in ATP content extracted from the cells were approximately two- to threefold compared to that of the starting samples, and the increase reached a maximum at 4 or 6 weeks after incubation. No significant ATP generation in M. leprae cells was demonstrated in medium at pH 6.2 or pH 6.6, in the original Kirchner medium with or without adenosine, or when cultured at 37 degrees C, or when containing an antileprosy drug. No detectable increase in the number of M. leprae cells was observed with the increase in intracellular ATP content and DNA replication. No effect was seen with renewal of the cultured medium by freshly prepared medium at 6 weeks' cultivation on the progressive ATP generation in M. leprae. PMID- 11480312 TI - PCR-restriction fragment length polymorphism analysis (PRA) of Mycobacterium leprae from human lepromas and from a natural case of an armadillo of Corrientes, Argentina. AB - Polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) which relies on the amplification of a 439-bp portion of the hsp65 gene present in all mycobacteria, followed by two distinct digestions (with BstEII and HaeIII) of the PCR product, offers a rapid and easy alternative that allows identification of the species without the need for specialized equipment. Wild leprosy in the nine-banded armadillo (Dasypus novemcinctus) is characterized by the presence of multiple bacilli in internal organs such as lymph nodes, spleen and liver, as well as in nerves and skin. We could observe this in 9 out of 132 animals captured in Corrientes, Argentina, an area endemic for leprosy in humans. Mycobacterium leprae were recognized in those naturally infected animals through different techniques. Three samples of extracted DNA of the mycobacteria present in the spleen, liver and popliteal lymph node of a naturally infected animal during the Experimental Program in Armadillo (PEA) and three samples of human lepromas were processed by PRA. The patterns of the six samples analyzed were identical and were characteristic of M. leprae. These studies, made for the first time in Argentina, corroborate the initial discoveries in South America made by our investigative group on the detection of armadillos naturally infected with the Hansen bacillus. PMID- 11480313 TI - Factors influencing the development of leprosy: an overview. AB - The clinical manifestations of leprosy vary, seemingly depending on the host's immune response. Mode and route of infection, such as skin versus nasal mucosa, insect bites, sexual and gastroenteral transmission, together with genetic factors that may contribute to the outcome of the infection, including HLA, Lewis factor, Nramp1 and more subtle inherited alterations, are discussed. It is theorized that a balance between host responses elicited by different routes of infection and size and spacing of inocula is responsible for the clinical and immunological manifestations of the disease. Genetic factors and contact with environmental microorganisms may modulate these responses. The final result, resistance, delayed-type hypersensitivity, tolerance, disease or no disease, spectrum and reactions, is most likely reached via the orchestration of the induced cyto- and chemokines. PMID- 11480314 TI - "Guide to eliminate leprosy as a public health problem" (first edition, 2000, World Health Organization). PMID- 11480315 TI - Facial lesions resembling leprosy. PMID- 11480316 TI - Leprosy case detection through community volunteers--a low cost strategy. PMID- 11480317 TI - Comparative characteristics of antigenic profile of M. leprae and M. lufu. PMID- 11480318 TI - Effects of vaccination with several mycobacterial proteins and lipoproteins on Mycobacterium leprae infection of the mouse. PMID- 11480319 TI - A study on the reproducibility of two serological assays for detection of Mycobacterium leprae infection. PMID- 11480320 TI - Peroxidase in therapy of experimentally induced leprosy. PMID- 11480321 TI - Histological resolution and bacterial clearance with pulse ROM therapy in borderline lepromatous leprosy. PMID- 11480322 TI - Does dapsone resistance really matter in the MDT era? PMID- 11480323 TI - "Metabolic theory of leprosy". PMID- 11480324 TI - Apoptosis in viral hepatitis B and C. AB - Tumor necrosis factor (TNF) and TNF receptors (TNFR) are members of the growing TNF ligand and receptor families known to be involved in apoptosis, viral pathogenesis and immune regulation. The present report will focus on the role of apoptosis in the pathogenesis of viral hepatitis B and C. Although TNF was reported years ago to modulate viral infections, recent findings on the molecular pathways involved in TNFR signaling have allowed a better understanding of the molecular interactions between cellular and viral factors within the infected cell. The interactions of viral proteins with intracellular components downstream of the TNFR have highlighted at the molecular level that viruses can manipulate the cellular machinery to escape the immune surveillance and to favor spread infection. We will review here the mechanism of apoptosis and the role of viral proteins that regulate apoptosis in viral hepatitis B and C. PMID- 11480325 TI - Extremely low sodium hypotonic rehydration solution for young children with acute gastroenteritis. AB - BACKGROUND: The clinical efficacy and safety of a low-sodium hypotonic oral rehydration solution (LSORS) was compared in a pilot study with that of a standard World Health Organization oral rehydration solution (STORS) in young children with acute diarrhea. METHODS: One hundred and seventeen boys aged 3 to 18 months with acute diarrhea were randomly assigned to groups and received low sodium (sodium 28 mmol/L) hypotonic oral rehydration solution and standard oral rehydration solution (sodium 90 mmol/L). Outcomes of therapy such as stool volume, duration of diarrhea and fluid intake were recorded at 24 hours, 48 hours, and the discontinuation of disease. Serum electrolytes levels were calculated before and after therapy. RESULTS: The stool output (gm/kg) in the first 24 hours was 69.2 +/- 38.5 in the LSORS group versus 105 +/- 39.8 in the STORS group (p < 0.05), while the total stool output during the 48-hour period was 151.4 +/- 68.9 in the LSORS group versus 232 +/- 97.3 in the STORS group (p < 0.05). No significant (p > 0.05) reduction in duration of diarrhea was noted (72.3 +/- 25.5 hours versus 69.8 +/- 29.4 hours). Four patients in the LSORS group and 7 patients in the STORS were classified as treatment failure. Forty four (80%) achieved successful treatment in the LSORS group and 34 (56.7%) in the STORS group (p = 0.07). CONCLUSION: LSORS is as safe as STORS in treating young children with acute gastroenteritis. No evidence of hyponatremia was found after patient received LSORS. LSORS afforded more efficacious therapy than STORS. PMID- 11480326 TI - Surgical treatment for pathological long bone fracture in patients with multiple myeloma: a retrospective analysis of 22 cases. AB - BACKGROUND: The purpose of this study was to retrospectively review the cases of pathologic long bone fractures caused by multiple myeloma treated in our hospital, to analyze the surgical method, complications, radiation therapy, survival time, and influence on quality of life. METHODS: In a retrospective study, 22 patients with the pathological long bone fractures due to multiple myeloma who were surgically treated between 1987 and 1997 were analyzed. All patients received open reduction and internal fixation either with plates or intra-medullary nailing. Cement augmentations were performed in the majority of cases (91%). A detailed retrospective analysis was done to correlate the surgical methods, radiation therapy, functional results, and complications post surgically. RESULTS: The most common site of fracture was the femur. The mean postoperative survival time was around 19 months. Post-operative pain relief was satisfactory, and only two patients required narcotics. No major complications were observed. However the union rate was only 30%, which might have been due to the inhibitory effect of radiation therapy on bone healing, or insufficient osteogentic ability of the myeloma-involved bone. CONCLUSION: Satisfactory pain relief and low implant failure rate was achieved and no definite evidence of tumor dissemination was found in this study. The authors suggest that open reduction and internal fixation with cement augmentation is a favorable treatment option for those patients suitable for surgery. However, postoperative radiation therapy may be associated with a low rate of union. PMID- 11480327 TI - Endoscopic hemoclip treatment of gastrointestinal bleeding. AB - BACKGROUND: Endoscopic hemostasis of gastrointestinal (GI) bleeding is a widely accepted modality of treatment, and endoscopic hemoclipping has been reported to cause fewer complications. METHODS: Forty patients with gastrointestinal bleeding (active bleeding or non-bleeding visible vessel), 30 men and 10 women with a mean age of 59.1 +/- 14.4 (28-86) years were treated with endoscopic hemoclipping. After panendoscopy or colonoscopy, a local epinephrine injection was routinely given in the initial 20 cases, followed by hemoclipping. For the latter 20 cases, local epinephrine was given only to those with active bleeding. If there was adherent blood clot, irrigation with 3% H2O2, and removal of the blood clot with forceps or basket were done. Six cases with bleeding at technically difficult locations were managed with a new method; a transparent cap (Olympus EMRC) fitted with a 2-channel endoscope for hemoclipping. RESULTS: There were 35 patients with peptic ulcer, 2 with post endoscopic papillotomy bleeding, 1 with duodenal Dieulafoy's lesion, 1 with Mallory Weiss syndrome, and 1 with rectal ulcer. The types of bleeding were spurting in 7, oozing in 12, and non-bleeding visible vessel in 21 cases. The average number of clips used was 3.1 +/- 1.7 (1-9) and the average clip loss was 0.6 +/- 0.9 (0-4) per patient. The success rate for hemostasis using the transparent cap-fitted endoscope was 100% and the overall success rate was 85% with no complications related to the procedure. CONCLUSION: Endoscopic hemoclip treatment for GI bleeding is safe and effective. The transparent cap-fitted endoscope is a new method for hemoclipping in technically difficult lesions. PMID- 11480328 TI - Arthroscopy in avulsion fracture of posterior cruciate ligament. AB - BACKGROUND: Surgical reduction of PCL avulsion fracture was performed from posterior approach and arthroscopic treatment was not routinely performed. However, associated intraarticular lesions of meniscus, cartilage, of any other pathologies were very common in PCL injuries. If the treatments of associated intraarticular lesions would improve the results was not known. Under this prospective study, we want to know if arthroscope is valuable in PCL avulsion fractures. METHODS: From January 1995 to December 1997, there were 29 cases of PCL avulsion fractures. They were grouped according to chart numbers. Group I was even number group and group II was odd number group. Arthroscopic examination was performed in even number group, but not in odd number group. Group I had 13 cases and group II had 16 cases. All the PCL avulsion fractures were repaired from posterior approach and fixed with cancellous screws. RESULTS: The range of motion, arthrometric data, and Lysholm score were similar in both group after more than 2 years of follow-up. The residual pain was less in arthroscopic group. Arthroscopic examination showed hemarthrosis in all cases of group I; meniscal lesion in 7 cases (54%), osteochondral or chondral lesions in 3 cases (23%), hard medial plica in 3 cases (23%), patellofemoral lesion in 1 case (7.7%), ACL tear in 1 case (7.7%), popliteal lesion in 2 cases (15.4%), lateral opening greater than 1 cm in 5 cases (38.5%). Though the functional results were similar in both groups, the arthroscopic findings in PCL avulsion fracture were negative in only 2 cases. CONCLUSION: Arthroscopic examination in PCL avulsion fracture can provide valuable information. PMID- 11480329 TI - Angiomyolipoma of the liver: case report. AB - Hepatic angiomyolipoma is a rare benign mesenchymal tumor of the liver. Most multiple hepatic angiomyolipomas have appeared in patients with renal angiomyolipoma and tuberous sclerosis. A 38-year-old female patient without chronic hepatitis B or C was hospitalized because of epigastric fullness for 2 months. Radiologic studies showed a large solid tumor with a small daughter nodule in the right hepatic lobe. Upon intravenous bolus injection of contrast medium, both tumors showed weak heterogeneous enhancement in the delayed phase. Although hepatocellular carcinoma was suspected by the findings of computed tomography, percutaneous transhepatic ultrasound-guided biopsy was performed for the large tumor. The histopathology showed many mature fat cells intermingled with thick-walled blood vessels, and epithelioid cells with eosinophilic cytoplasm; the epithelioid cells stained positively for HMB-45 and smooth muscle actin. Angiomyolipoma of the liver was confirmed. The main tumor enlarged considerably during a follow-up period of 3 years. Surgical resection was performed due to persistent symptoms. She had an uneventful postoperative recovery and was well when followed up 10 months after surgery. We should be aware that a hepatic angiomyolipoma can change in size during its natural course, and this finding does not necessarily indicate malignancy. PMID- 11480330 TI - Chylous ascites in acute pancreatitis during pregnancy: case report. AB - Chylous ascites is a rare clinical manifestation characterized by ascitic chylomicrons resulting from mechanical obstruction of or leakage from the lymphatic channel. Chronic disorders, especially malignancies, account for most cases of chylous ascites. Acute chylous ascites is less common than the chronic form. We present a rare case of acute chylous ascites secondary to acute pancreatitis during the third trimester of pregnancy. This 24-year-old woman was referred to our emergency department because of severe epigastralgia for several days. Abdominal computed tomography revealed diffuse enlargement of the pancreas and peripancreatic exudation. Massive chylous ascites was found during emergent abdominal exploratory laparotomy. An emergent cesarean section was done because of fetal distress and there was no further accumulation of chyle. A pancreaticocutaneous fistula resulting from the cesarean section was treated successfully with a fistulectomy. In conclusion, chylous ascites is a rare complication of acute pancreatitis. Cesarean section may be helpful in terminating chylous accumulation in acute pancreatitis during the third trimester of pregnancy. PMID- 11480331 TI - Progression of hydroxychloroquine retinopathy after discontinuation of therapy: case report. AB - Chloroquine and its derivative, hydroxychloroquine sulfate, have been used in treating malaria, dermatitides of systemic lupus erythematosus and rheumatoid arthritis. Hydroxychloroquine retinopathy is uncommon in Taiwan. Here we report a patient with hydroxychloroquine retinopathy which progressed even after discontinuation of hydroxychloroquine. A 42-year-old woman had systemic lupus erythematosus for twenty years. She had been treated with 200 to 400 mg of hydroxychloroquine per day (4 to 8 mg/kg of body weight/day) with a cumulative dose of 657 g. After bull's-eye maculopathy was found, hydroxychloroquine was discontinued. Her medical history revealed no chloroquine administration and no other systemic disease. Five years after cessation of the therapy, her visual acuity and visual fields continued to deteriorate. Ophthalmoscopic examination revealed the hydroxychloroquine retinopathy had advanced. To the best of our knowledge, the progression of hydroxychloroquine retinopathy after discontinuation of medications is a rare phenomenon. Regular ophthalmologic examinations should be performed for patients on hydroxychloroquine regimens because there is no satisfactory treatment for hydroxychloroquine retinal toxicity. Ophthalmologists, dermatologists and rheumatologists should monitor for ocular toxicity of hydroxychloroquine carefully. PMID- 11480332 TI - Combined computed tomographic pulmonary angiography and venography for evaluation of pulmonary embolism and lower extremity deep venous thrombosis: report of two cases. AB - Pulmonary embolism (PE) and deep venous thrombosis (DVT) are major causes of morbidity and mortality, which can be reduced with accurate diagnosis and proper treatment. More than 90% of PEs originate in lower-extremity DVT. Currently, evaluation of PEs and lower-extremity DVT requires 2 separate tests (ventilation perfusion scan, computed tomographic pulmonary angiography (CTPA), or pulmonary angiography for PE and sonography, computed tomographic venography (CTV), conventional venography, or magnetic resonance venography for DVT). Combined computed tomographic pulmonary angiography and venography (CTPAV) is a new diagnostic technique that combines CTPA and CTV into a single study for the screening of PE and subdiaphragmatic DVT. CTPAV is a modified CTPA study that evaluates the subdiaphragmatic deep vein system at the time of CTPA, without additional venipuncture or contrast medium. It is easy to perform, fairly easy to interpret, readily available, and requires no invasive procedure. We present 2 cases of multiple PE and lower-extremity DVT in which CTPAV was used. PMID- 11480333 TI - Differentiation of breast calcifications. AB - Breast calcifications display a variety of characteristics, depending on the physiological process leading to their development. Because certain types of calcifications can be indicative of breast cancer, it is important that mammographers recognize which calcifications signify a benign or malignant process. In addition to discussing the causes of breast calcifications and describing ways to optimize their mammographic depiction, this article explains the differences in the mammographic appearances of various types of breast calcifications. PMID- 11480334 TI - Cadaver imaging as a learning alternative. AB - The radiologic science department at Armstrong Atlantic State University in Savannah, Ga, incorporated cadaver radiography into its educational program. This article discusses the benefits and limitations of using cadavers in radiography education, as well as the curricular, physical and psychological issues involved. PMID- 11480335 TI - Positron emission tomography. AB - Positron emission tomography is a sophisticated, physiology-based imaging technique that provides information about the function of tissues and organs. Combining PET data with computed tomography or magnetic resonance images provides clinicians with physiological information linked to an anatomical site. This overview discusses the biological principles underlying the technology, PET radiopharmaceuticals, PET imaging facilities, specific imaging applications and reimbursement issues. PMID- 11480336 TI - Neonatal imaging. AB - This article reviews diagnostic imaging of newborns, beginning with a discussion of helpful approaches and safety concerns and an overview of imaging techniques used. The author then presents a variety of common neonatal disorders and their radiologic appearance. PMID- 11480337 TI - New standards. PMID- 11480338 TI - Sensitivity numbers in chest CR. PMID- 11480339 TI - Breast hamartoma. PMID- 11480340 TI - Rings and things. PMID- 11480341 TI - Integrating theory and practice. PMID- 11480342 TI - Patient page. Nuclear medicine imaging. PMID- 11480343 TI - Methods of statistical analysis of quantitative data obtained by toxicological bioassays using rodents in Japan: historical transition of the decision tree. AB - Massive amounts of quantitative data are being obtained by toxicity tests with time. These data sometimes include the data with a different distribution pattern or with different numbers of animals in each group. The tree-type algorithm has long been used commonly to promptly analyze the difference between the control group and dosage groups. In Japan, the decision tree has been used since 1982 with several modifications of the initial procedure. Recently, the method has been revised to have a high power. The changes have been made in two directions; one is a sophisticated method and the other is a simplified method. In this paper, the historical transition of the decision tree is described and the future of the decision tree forecasted. PMID- 11480344 TI - Antifeedant activity of aqueous extract of Gnidia glauca Gilg. and Toddalia asiatica Lam. on the gram pod borer, Helicoverpa armigera (Hbn). AB - Aqueous leaf extracts of two plants namely Gnidia glauca Gilg. and Toddalia asiatica Lam., have been screened for their antifeedant activity against the sixth instar larvae of Helicoverpa armigera (Hbn) by applying the aqueous leaf extracts at various concentrations viz., 0.2, 0.4, 0.6, 0.8 and 1.0 percent on young tomato leaves. The larval mortality of more than 50 percent at higher concentrations (0.8 and 1.0 percent) was observed in the aqueous extracts. Among the two aqueous leaf extracts tested, T. asiatica was found to show higher rate of mortality (86.1%) at 1.0 percent concentration. A reduction in the rate of food consumption and growth was observed in the larvae of H. armigera after 48 hours of treatments in both the aqueous extracts. Since this insect pest species have developed resistance and resurgence to synthetic insecticides, the only alternate is the usage of bio-pesticides for they are eco-friendly, pollution free and easily degradable. PMID- 11480345 TI - Effect of mercuric chloride on testicular activities in mice, Musculus albinus. AB - Impaired testicular function was observed after an exposure of Swiss albino mice (30 +/- 2 g) to mercuric chloride. A sublethal chronic exposure (0.5 ppm for 21 days) resulted in regressed histological and histochemical properties of the testis. The changes observed were degenerated tunica albuginea, abnormal configurations of seminiferous tubules, deformed primary and secondary spermatocytes, hypertrophy and vacuolization in interstitial cells and Sertoli cells. The 3 beta and 17 beta hydroxy steroid dehydrogenase enzyme and the level of testosterone hormone were significantly (p < 0.001) reduced. The diameter of different spermatogenic cells were significantly (p < 0.001) reduced. PMID- 11480346 TI - Residual toxicity of Cypermethrin in the larvae of coconut pest Oryctes rhinoceros (Coleoptera: Scarabaeidae). AB - The toxic effect of Cypermethrin 10 EC (0.125, 0.25 and 0.5%) was estimated in the bodywall and digestive system of the larvae of Oryctes rhinoceros by HPLC after exposing them to different concentrations (0.125, 0.25 and 0.5%). Among the various concentrations used maximum residues were detected in bodywall (0.25%), whereas at higher concentration (0.5%) the residue detected was minimum. The treatment of Cypermethrin was found to be highly toxic upto 12 h of treatment, after which it declined reaching the minimum by 24 h. The residue of Cypermethrin could not be detected in digestive system. The experiments indicate the pesticide get concentrated in the bodywall to a maximum extent. PMID- 11480347 TI - Microbial decolourisation of pulp and paper mill effluent in presence of nitrogen and phosphorus by activated sludge process. AB - The effect of pH, nutrient and aeration was studied on the removal of colour and reduction of BOD, COD and heavy metals with addition of readily available source of nitrogen and phosphorus in concentration of 1.0 g/L. Recalcitrant compound was effectively degraded by active microbial consortia. The isolated bacteria were identified as Pseudomonas putida (S1), Citrobacter sp. (S4) and Enterobacter sp. (S5). These organisms not only decolourised effluent upto 97% but reduced BOD, COD, phenolics and sulfide upto 96.63, 96.80, 96.92 and 96.67% respectively within 24 hrs of aeration and the heavy metals were removed upto 82-99.80%. The TSS and TDS were sharply reduced due to degradation. The absorption maxima was also decreased to 90%. However, in control without the microbial consortium no noticeable change was produced. PMID- 11480348 TI - Impacts of crude oil on the germination and growth of cress seeds (Lepidium sp.) after bioremediation of agricultural soil polluted with crude petroleum using "adapted" Pseudomonas putida. AB - The impacts of crude oil on the germination, growth and morphology of cress seeds (Lepidium sp.) after bioremediation of agricultural soil polluted with crude petroleum using "adapted" Pseudomonas putida (PP) were examined for 15 days. At day 15 there was 100% germination in the untreated control samples, the mean height of the seedlings was 75.8 +/- 2.6 mm and all appeared to have grown morphologically normal. In the experimental samples treated with oil and PP inoculation, there was 98% germination and the seedlings reached a height of 63.8 +/- 6.9 mm; again, morphologically the seedlings appeared normal. However, in the control samples treated with oil but without PP inoculation, there was 31-38% germination and seedling heights of 34.2 +/- 11.4-42.3 +/- 8.5 mm with abnormal morphology. Treatment of oil-impacted agricultural soil with PP as a bioremediation agent does produce soil which is capable of growing larger and healthier plants than where bioremediation has not taken place. PMID- 11480349 TI - Biochemical responses of Camellia sinensis (L.) O. Kuntze to heavy metal stress. AB - Three heavy metals-mercury (II), copper (II) and nickel (II), each at a concentration of 10 and 100 micrograms/ml, were tested for their effects on various biochemical constituents of tea leaves. Both NI (II) and Hg (II) decreased the phenolic contents, while Cu (II) increased it to some extent. The metal treatments enhanced the activity of phenyl alanine ammonia lyase (PAL), while the activity of poly phenol oxidase (PPO) showed a decline. Heavy metal stress also decreased the chlorophyll content of the leaves, along with a significant reduction in Hill activity. Proline content increased significantly in all treatments. PMID- 11480350 TI - A study of Malaiyur village ecosystem of Madurai district, Tamil Nadu (India). AB - The interaction between man and environment in Malaiyur, a hilly village of Madurai District (Tamil Nadu) was studied during the year 1998. The village is sustained on agriculture labour and hence occupies the lowest hierarchy with traditional occupation and the people may be categorised as 'ecosystem people'. The study suggests that the people has to be changed as 'Biosphere people' to protect the environmental degradation due to human activities. PMID- 11480351 TI - Evaluation of impact of sewage irrigation on cytotoxicological potentialities of Chenopodium album in Allium assay. AB - Chenopodium album is a weed commonly consumed by North Indian population as vegetable. Plants are known to accumulate toxins from their environment. Presently, the leaf homogenates of these plants growing in a tubewell irrigated field and a sewage irrigated field were evaluated for cytotoxicological effects in Allium root tip assay. Studies revealed that Chenopodium album was mildly mitodepressive in nature and was capable of inducing chromosomal aberrations. The leaf homogenate of the plants growing in sewage irrigated fields induced more quantum of aberrations than the plants from the control site. This has a direct bearing on consumability of sewage grown vegetables and fodder. PMID- 11480352 TI - Effect of Aegle marmelos and Hibiscus rosa sinensis leaf extract on glucose tolerance in glucose induced hyperglycemic rats (Charles foster). AB - In an effort to test the hypoglycemic activity of Aegle marmelos and Hibiscus rosa sinensis in glucose induced hyperglycemic rats, their alcoholic leaf extracts were studied. Both the groups of animals receiving either. A. marmelos or H. rosa sinensis leaf extract for seven consecutive days, at an oral dose equivalent to 250 mg kg-1 showed significant improvements in their ability to utilize the external glucose load. Average blood glucose lowering caused by A. marmelos and H. rosa sinensis was 67% and 39% respectively, which shows that former significantly (p < 0.001) improves the glucose tolerance curve. The magnitude of this effect showed time related variation with both the plants. Efficacy of A. marmelos and H. rosa sinensis was 71% and 41% of glybenclamide, respectively. These data throw some light on the possible mechanism of hypoglycemic activity of both the plants. The mechanism of action could be speculated partly to increased utilization of glucose, either by direct stimulation of glucose uptake or via the mediation of enhanced insulin secretion. PMID- 11480353 TI - Effect of textile industrial effluent on tree plantation and soil chemistry. AB - A field study was conducted at Arid Forest Research Institute to study the effect of textile industrial effluent on the growth of forest trees and associated soil properties. The effluent has high pH, electrical conductivity (EC), sodium adsorption ratio (SAR) and residual sodium carbonate (RSC) whereas the bivalent cations were in traces. Eight months old seedlings of Acacia nilotica, Acacia tortilis, Albizia lebbeck, Azadirachta indica, Parkinsonia aculeata and Prosopis juliflora were planted in July 1993. Various treatment regimes followed were; irrigation with effluent only (W1), effluent mixed with canal water in 1:1 ratio (W2), irrigation with gypsum treated effluent (W3), gypsum treated soil irrigated with effluent (W4) and wood ash treated soil irrigated with effluent (W5). Treatment regime W5 was found the best where plants attained (mean of six species) 173 cm height, 138 cm crown diameter and 9.2 cm collar girth at the age of 28 months. The poorest growth was observed under treatment regime of W3. The growth of the species varied significantly and the maximum growth was recorded for P. juliflora (188 cm height, 198 cm crown diameter and 10.0 cm collar girth). The minimum growth was recorded for A. lebbeck. Irrigation with effluent resulted in increase in percent organic matter as well as in EC. In most of the cases there were no changes in soil pH except in W5 where it was due to the effect of wood ash. Addition of wood ash influenced plant growth. These results suggest that tree species studied (except A. lebbeck) can be established successfully using textile industrial wastewater in arid region. PMID- 11480354 TI - Hydrobiological study of Dahikhuta reservoir. AB - Physico-chemical parameters and the algae of Dahikhuta reservoir, near Malegaon, Dist. Nasik (Maharashtra) have been investigated during July to December 1998. The results have revealed that reservoir water is classified as oligotrophic on the basis of water quality criteria. This paper also reports algal diversity. Total 19 algal taxa are reported from three classes. PMID- 11480355 TI - Determination of pesticide residues in fruits and vegetables in Islamabad market. AB - Pesticide residues in fruits and vegetable from Islamabad market, Pakistan were determined by GLC using Perkin Elmer Autosystem equipped with 63NECD and capillary column. Dimethoate was determined in the quantity of 0.032 mg kg-1 in apple, 0.110 mg kg-1 in banana, 0.004 mg kg-1 in brinjal, 1.80 mg kg-1 in cauliflower and 0.13 mg kg-1 in arvi, fenvalerate 0.010 mg kg-1 in apple and chlorpyrifos 0.004 mg kg-1 in brinjal. The importance of these pesticide residues with reference to human health is also briefly discussed. PMID- 11480356 TI - Effect of trypanosomes infection on blood ascorbic acid and serum aldolase levels on the fresh water fishes, Clarias batrachus and Heteropneustus fossilis. AB - The trypanosomes presence has been found to decrease blood ascorbic acid levels in fishes, Clarias batrachus and Heteropneustus fossilis by 55.7% and 54.70% respectively. The infection also showed significant increase in serum aldolase level of three fishes by 50.19% and 48.06% respectively. PMID- 11480357 TI - Environmental education & research have attained a level of respectability. PMID- 11480358 TI - Integumental disease in invertebrates. AB - The necessarily truncated discussion of integumental neoplasia in invertebrates, given the lack of investigation of such lesions involving individual animals, shows how invertebrate veterinary medicine has a long way to go before reaching the levels of diagnosis and therapeutics of vertebrate exotic species. Nevertheless, the importance of a disease such as varoosis in a species such as the honey bee, which has a massive influence through pollination quite beyond its relevance as a honey producer, shows the importance of never excluding invertebrates from discussion of disease in exotic species. It is therefore commendable that this issue of the Veterinary Clinics of North America: Exotic Animal Practice seeks wherever possible to include invertebrates in its discussion. PMID- 11480359 TI - Cutaneous biology and diseases of fish. AB - The normal structure and function of the piscine integument reflects the adaptation of the organism to the physical, chemical, and biological properties of the aquatic environment, and the natural history of the organism. Because of the intimate contact of fish with the environment, cutaneous disease is relatively more common in fish than in terrestrial vertebrates and is one of the primary disease conditions presented to the aquatic animal practitioner. However, cutaneous lesions are generally nonspecific and may be indicative of disease that is restricted to the integument or a manifestation of systemic disease. Regardless, a gross and microscopic examination of the integument is simple to perform, but is highly diagnostic and should always be included in the routine diagnostic effort of the aquatic animal practitioner, especially since various ancillary diagnostic procedures are either not practical or lack predictive value in fish. The purpose of this article is to provide an overview of normal cutaneous biology prior to consideration of specific cutaneous diseases in fish. PMID- 11480360 TI - Amphibian skin diseases. AB - Skin diseases in amphibian species seem to carry an additional degree of seriousness compared with those of other vertebrates. Because of the skin's importance in respiration and ion transport, breaching of the integument of these animals can result in fatal septicemias or metabolic disturbances. The timely diagnosis and treatment of these skin lesions is important. A review of the diseases affecting amphibian skin is provided. This article also describes the clinical signs, appropriate diagnostic steps, and treatment and control of amphibian skin diseases. PMID- 11480361 TI - Dermatology of reptiles: a clinical approach to diagnosis and treatment. AB - As herpetoculture becomes more commonly practiced, many of the dermatologic problems discussed in this article will be encountered less frequently. Many of the problems result from a general lack of knowledge regarding proper nutrition, husbandry, and the overall needs of reptiles within a captive environment. As reptiles become more a part of the mainstream pet population, information about their care will undoubtedly become more accessible, and as more captive-raised reptiles become available in the pet trade, many of the dermatoses currently associated with wild-caught animals will become less common. PMID- 11480362 TI - Avian skin diseases. AB - Avian dermatology is an important part of avian practice. Feather plucking, or self-mutilation, is one of the most common and challenging syndromes seen in avian practices, yet our understanding of it has developed piecemeal. Psittacine circovirus, which causes psittacine beak and feather disease, can devastate breeding programs and cause masked distress to new bird owners and their young birds. Cutaneous neoplasms are increasing in incidence as veterinarians are seeing more older bird patients. PMID- 11480363 TI - Skin diseases of rodents and small exotic mammals. AB - Small exotic mammals and rodents are becoming popular pets in the United States. Like most other exotics, the popularity of these animals has vastly preceded the accumulation of practical husbandry and veterinary information available about them. Several dermatologic conditions have been described in most rodents and small exotic mammals; however, the practitioner can assume that more exist that have not yet been diagnosed or documented. It is not unreasonable to assume that rodents and small exotic mammals could be affected by many of the same dermatologic conditions well described in other animals. Veterinarians are encouraged always to apply the same diagnostic protocols used to work up skin problems in dogs and cats when presented with an exotic pet with a dermatologic disease. PMID- 11480364 TI - Skin disorders of the rabbit. AB - The integument is a common site of disease in the rabbit. Parasites, bacterial infections, and viral infections predominate; however, trauma, behavioral, and neoplastic diseases are common as well. Wild rabbits act as the reservoir of many parasitic and viral diseases of domestic rabbits. PMID- 11480365 TI - Skin diseases of ferrets. AB - This article describes the most common diseases that affect the integumentary system of ferrets. Emphasis is placed on diagnosis and treatment of each disease. PMID- 11480366 TI - Nonhuman primate dermatology. AB - Clinical skin disease in the NHP is common and generally similar in appearance, clinical approach, and therapy to that seen in other animals and humans. Because most skin diseases of NHP are potentially zoonotic, care must be taken when handling the animals. Traumatic injury to the skin with secondary bacterial infection is the most common condition seen clinically. Virus-induced skin disease is probably the next most commonly seen, followed by mycotic infections, parasitisms, cancer, and miscellaneous rare conditions. PMID- 11480367 TI - [A fast algorithm for homogeneous Ir-192--afterloading irradiation of cylindrical surfaces]. AB - A fast algorithm was required to homogenise the dose on a cylinder mantle around a single straight applicator for Ir-192 afterloading. For linear applicators, an approximation formula for homogenisation of dose on one arbitrary cylinder mantle around the catheter is derived that predicts the required rise of dwell time tEnd at the end of the applicator. The other positions are loaded with a constant dwell time tMitte. The dwell time at the end tEnd appears to be independent of applicator length and can be described as a function of tMitte. In contrast to homogeneous loaded applicator, the dwell time in the central positions becomes also almost independent of catheter length, and is strictly proportional to the optimisation radius. TLD measurements and comparisons with a commercial planning system show the applicability of the procedure. This guarantees a homogeneous dose distribution even when modifications of the application length or optimisation distance become necessary. The algorithm can also be used to control the results of a planning system independently. PMID- 11480368 TI - On small angle multiple coulomb scattering of protons in the gaussian approximation. AB - Experimental data from the literature on small-angle multiple Coulomb scattering of protons in various materials were analysed in order to device an equation for the scattering angle in the Gaussian approximation. In comparison to Highland's well-known formula, the present approximation can be integrated to take into account energy loss in the scattering media. In addition, it is more precise than Highland's formulation for thin and thick scatterers consisting of elements with low atomic number. The simple equation obtained in this study can be used to obtain prompt answers for scattering problems which can occur, for example, in proton therapy or proton radiography. PMID- 11480369 TI - [EGs-Ray, a program for visualization of Monte Carlo calculations in radiation physics]. AB - A Windows program is introduced which allows a relatively easy and interactive access to Monte Carlo techniques in clinical radiation physics. Furthermore, this serves as a visualization tool of the methodology and the results of Monte Carlo simulations. The program requires only little effort to formulate and calculate a Monte Carlo problem. The Monte Carlo module of the program is based on the well known EGS4/PRESTA code. The didactic features of the program are presented using several examples common to the routine of the clinical radiation physicist. PMID- 11480370 TI - [Influence of tandem optics and television tube of modern digital fluoroscopy equipment on image quality]. AB - The influence of the tandem optics and the TV-tube to the image quality of 1k- and 2k-radiographs of a modern fluoroscopy equipment should be assessed on the basis of objective parameters and clinical requirements. Radiographs of different phantoms with 1k- and 2k-image matrix, different illumination point corrections (IPC) and signal gray levels were taken, examined and evaluated, in order to determine the objective image parameters and the clinical relevancy of these parameters. On the one hand, the digital images data could be used directly for evaluation; on the other hand, the radiographs could be visually evaluated by experienced radiologists within the framework of a blind study. The IPC is controlled by the aperture of the iris diaphragm and the insertion of a neutral gray filter in the tandem optics. The larger the aperture of the iris diaphragm (at constant image receiver dose) the higher were gradation, signal to noise ratio (SNR) and image homogeneity. Furthermore, the larger the aperture, the lower was the square wave response function (SWRF). The insertion of a gray filter in the tandem optics decreases gradation, SNR and homogeneity, and improves the SWRF. PMID- 11480371 TI - Film dosimetric verification of the Voxel Monte Carlo (VMC) algorithm with electron beam dose distributions. AB - Monte Carlo (MC) methods have the potential to predict radiation-therapy doses more accurately than any conventional technique, but normal MC simulations are very time consuming. Therefore, a fast MC code (Voxel Monte Carlo; VMC) was developed especially for radiation therapy purposes and experiments with the comparable precision were performed to demonstrate its accuracy. In the present study the dose distributions measured with film dosimetry in a cylindrical phantom were compared with calculations derived by VMC. The phantom consisted of 18 circular shaped PMMA slabs with a diameter of 20 cm and a thickness of approx. 1 cm. The films were placed between the slabs, and the whole phantom was irradiated with electron beams of different energies (6 MeV, 10 MeV, 18 MeV). The measured optical density distributions were then converted into dose distributions using characteristic curves of the film. Taking into account experimental uncertainties and statistical calculation fluctuations, agreement was found between measurements and VMC simulations with a maximal deviation of 3 mm on isodose curves for 18 MeV. PMID- 11480372 TI - [Radiation exposure in diagnostic radiation: a quantification model for analytic epidemiological studies]. AB - The present paper introduces a model for retrospective assessment of radiation exposure to patients undergoing conventional diagnostic radiography. The model allows for the quantification of relevant organ doses as well as the effective dose from exposures in the time period between 1946 and 1995. The assessment procedure consists of three consecutive steps. The first step uses a set of ideal doses that were measured under optimum conditions. In the second step, we account for the state of radiological technology available at the time of a given exposure. The third step, finally, considers the prevailing standard of radiological practice. Model parameters are derived from a compilation of data from the contemporary radiologic literature as well as from a multitude of historic sources. The three-dimensional matrix of ideal doses and respective correction factors presented here can be readily implemented in the context of analytic epidemiological studies. PMID- 11480373 TI - ECG of the month. Outbursts. Paroxysmal atrial fibrillation. PMID- 11480374 TI - Radiology case of the month. An 11-year-old girl with a pulse discrepancy between the upper and lower extremities. Coarctation of aorta. Widening of the radioulnar joint. PMID- 11480375 TI - Three resident cases at Charity Hospital. PMID- 11480376 TI - Acute coronary syndromes: the American epidemic continues. PMID- 11480377 TI - Epidemiology and pathophysiology of acute coronary syndromes. AB - In the past 2 decades great strides have been made in the understanding of the physiology of acute ischemic coronary syndromes. The description of acute plaque rupture and thrombus formation has enabled improved treatments and outcomes. Despite a decrease in death rates from ischemic syndromes, the total number of deaths continues to increase due to population changes. This paper will characterize the problem and review current understanding of acute ischemic physiology. PMID- 11480378 TI - Medical management of acute ST elevation myocardial infarction. AB - Advances have occurred rapidly in how the patient who presents with acute ST elevation myocardial infarction is managed. Nevertheless, due to availability and effectiveness, medical therapy continues to be the mainstay of treatment. Several drugs are of proven efficacy in decreasing myocardial loss and mortality. Thrombolytics, antiplatelets, and anticoagulants combine to decrease the thrombus overlying the active plaque and thus increase blood flow to the infarcting myocardium. Beta blockers, nitrates, and angiotensin converting enzyme inhibitors diminish myocardial oxygen consumption and have been shown to decrease morbidity and mortality. Supportive therapies provide patient comfort and aid survival until definitive therapies are instituted. However, as advanced as treatments become, none are effective unless patients are educated to seek help quickly and emergency response systems are efficient. Health care providers must ensure patient education and prompt medical response in communities so that medical regimens can be as effective as possible. PMID- 11480379 TI - Interventional therapy for acute myocardial infarction. AB - Percutaneous transluminal coronary angioplasty has proven to be more effective and safer than thrombolytic therapy for the treatment of acute ST elevation myocardial infarction. Coronary intervention decreases early mortality and the incidence of intracerebral hemorrhage when performed by an experienced interventional team in a timely fashion. After failed fibrinolytic therapy for myocardial infarction, percutaneous transluminal coronary angioplasty is indicated for signs of ischemia and is very effective in restoring vessel patency and reducing mortality when used as a rescue procedure. The glycoprotein IIb-IIIa inhibitors improve outcomes in percutaneous transluminal coronary angioplasty, particularly in patients undergoing stent placement. Percutaneous interventional therapy in acute myocardial infarction is particularly beneficial in patients with cardiogenic shock and effective for saphenous vein graft occlusions, patients with diabetes mellitus, and in the elderly. New devices and drugs are currently being tested for acute myocardial infarction and provide hope for even better interventional therapies in the near future. PMID- 11480380 TI - Management of cardiac arrhythmias in acute coronary syndromes. AB - A variety of atrial and ventricular arrhythmias are encountered in patients with acute coronary syndromes. These include both brady and tachyarrhythmias of supraventricular and ventricular origin. Sinus bradycardia and atrial fibrillation are the most common of supraventricular origin. Ventricular arrhythmias that merit consideration include premature ventricular complexes, accelerated idioventricular rhythm, ventricular tachycardia, and ventricular fibrillation. Intraventricular and atrioventricular conduction blocks associated with acute coronary syndromes include bundle branch blocks, fascicular blocks, and various degrees of atrioventricular block. A review of management of these arrhythmias is presented. PMID- 11480381 TI - Management of unstable angina and non-ST-segment elevation myocardial infarction. AB - Most of the patients with acute coronary syndromes present with unstable angina or non-ST-segment elevation myocardial infarction. Recent advances in the diagnosis and treatment of these conditions have changed the management guidelines, allowing for better patient care. The purpose of this review is to summarize the current knowledge and present both established and new or evolving strategies used in the treatment of patients with unstable angina and non-ST segment elevation myocardial infarction. Use of aspirin, nitrates, beta-blockers, calcium channel blockers, and anticoagulation with unfractionated heparin has been a common practice in these patients. While still effective and recommended, the emergence of new anticoagulants (low molecular weight heparins) and antiplatelet agents (ADP inhibitors and glycoprotein IIb/IIIa blockers) has added significant benefit to the treatment options and has resulted in a better prognosis for the patients. It also appears that, in view of recent trials, an early invasive approach is becoming the standard of care, especially for high risk patients. PMID- 11480383 TI - Battling Osler's great foes: apathy, ignorance, and poor quality indicators. PMID- 11480382 TI - Risk reduction strategies in coronary artery disease. AB - The growing knowledge of the basic science of the atherosclerotic plaque, the identification of modifiable risk factors, and the development of the effective medical therapies have provided the physician with powerful tools to alter the course of atherosclerotic disease in the post myocardial infarction patient. Lipids have emerged as a primary target for modification in addition to attention to traditional lifestyle modification such as smoking cessation, diet, and exercise. This article reviews lipid management as well as therapy and goals. Traditional and some "new" risk factors are also addressed. The risk of subsequent events in the post myocardial infarction patient can be dramatically altered, thus the patient who follows suggested guidelines can expect to have improved quality and quantity of life after an infarction. PMID- 11480384 TI - Prescription abuse can no longer be ignored. PMID- 11480386 TI - A selection of trends in HIV research. PMID- 11480387 TI - Projected benefits of molecular screening for hepatitis C in an underdiagnosed rural U.S. population. AB - In summary, the main goal of the proposal is to detect HCV infection in a rural population and the subsequent quantification of viral load in HCV-positive subjects so that proper intervention therapies can be initiated as early as possible postinfection. An additional benefit of this project is the potential to diffuse new technology into rural health care. This article is designed to elicit feedback and comments towards the feasibility and benefits of molecular screening in a rural community in Arizona through the project proposed by the author. PMID- 11480388 TI - Alcohol use and HIV infection in the HAART era. PMID- 11480389 TI - Use of bleach to disinfect HIV-1 contaminated syringes. PMID- 11480390 TI - Tartrate-resistant acid phosphatase as a serum marker of bone resorption. PMID- 11480391 TI - The American Society for Microbiology. PMID- 11480392 TI - Combined behavioral and pharmacologic treatments of alcoholism. PMID- 11480393 TI - Changing face of medical practice in the millennium as occurred in India. PMID- 11480394 TI - Novel therapies for the acute respiratory distress syndrome. PMID- 11480395 TI - A comparative of the effect of tamoxifen on serum lipid and lipoprotein profile in premenopausal & postmenopausal women with breast carcinoma & associated risk of vascular complication. AB - To examine the effect of tamoxifen on serum lipid and lipoprotein profile of premenopausal and postmenopausal patients of breast carcinoma (with and without vascular disease) we performed a short term evaluation of serum lipid and lipoprotein profile of 38 pre and 42 post menopausal subjects of breast carcinoma (with and without vascular disease) at baseline and during 3 and 6 months of tamoxifen therapy. The lipid & Lp profile of premenopausal patients of Breast carcinoma without vascular disease showed no significant variation after 3 and 6 months of tamoxifen treatment than the corresponding baseline values, but in premenopausal subjects of breast cancer with vascular disease and post menopausal subjects of breast carcinoma (both with and without vascular disease), Serum TC, Apo-B and Lp (a) were significantly decreased and serum HDL and Apo A-I were elevated significantly, However serum TG remain unaltered in premenopausal patients of breast carcinoma with vascular disease but found to be elevated significantly in postmenopausal subjects of breast cancer, both, with and without vascular disease, after 3 and 6 months of tamoxifen treatment, than the corresponding baseline values. Also, the comparison of the results of the present study for pre and postmenopausal patients of breast cancer revealed that the administration of tamoxifen, as adjuvant therapy for breast cancer, is beneficial for postmenopausal patients of breast carcinoma with vascular disease as the drug minimises the risk of vascular disease by bringing significant improvement in serum lipid and Lp profiles of the patients but does not have any significant beneficial effect on familial hyperlipidemic patients of breast cancer. PMID- 11480396 TI - Seasonal variations in incidence of severe and complicated malaria in central India. AB - The geographical position and climate of India is favorable for the transmission of malarial infection. The maximum prevalence of malaria in most parts of India is from July to November months. Rainfall provides mosquitoes, a breeding ground giving rise to epidemics. We studied the seasonal variation in cases of severe and complicated malaria presenting at MGIMS, Sevagram, Wardha (Vidarbha region in Maharashtra) over a period of three years. The findings of peak of malaria observed during September-November during three years period points to the fact that the increase in vector breeding after rainy season is responsible for the upsurge in the malarial cases during these months. This also indicates that this area (Vidarbha) has an unstable transmission of malaria. PMID- 11480397 TI - Lacunar syndromes due to small haemorrhagic lesions. AB - OBJECTIVE: To study the frequency, clinical presentations and prognostic aspects of haemorrhagic lacunar syndromes. MATERIAL AND METHODS: 375 patients of cerebrovascular accidents admitted in neurology wards over 2 years were studied. Based on CT findings, 114 patients were diagnosed as intra cerebral haemorrhage (ICH). They were assessed in detail by careful history and examination of neurological status. Coagulation studies, MRI and angiography were done in selected cases. RESULTS: In 17 (15%) patients, size of hematoma was < 2 cms as seen in CT scan. The sites of these small haemorrhages were in basal ganglia (6 patients), internal capsule (3 patients), thalamus (5 patients) and pons (3 patients). Depending on their location they presented as pure motor stroke, pure sensory stroke, ataxic hemiparesis, sensorimotor stroke and clumsy hand-dysarthia syndrome. Three patients showed interesting phenomena like transient ischemic attack caused by bleed or previous disease showing improvement after bleed. Prognosis of these small ICH was excellent with 100% survival rate and lesser disability as shown by good functional recovery. CONCLUSIONS: Haemorrhagic lacunar syndromes are not just the statistical curiosities but could have significant prognostic bearing. PMID- 11480398 TI - Haemangiopericytoma--an unusual tumour--a report of 5 cases. AB - Haemangiopericytoma (HPC) has remained a controversial entity ever since its first description. The main histological criterion to diagnose this entity is the characteristic vascular pattern highlighted by the reticulin stain. Since this pattern can be present in several other connective tissue tumours, it is basically a diagnosis of exclusion. Electron microscopy has however been helpful in some cases. PMID- 11480399 TI - Comments on "Predicted region of stability for balance recovery: motion at the knee joint can improve termination of forward movement". PMID- 11480400 TI - EUFEPS conference report. Optimising drug development: strategies to assess drug metabolism/transporter interaction potential - towards a consensus. European Federation of Pharmaceutical Sciences. PMID- 11480402 TI - Coping with the stress of malpractice litigation. PMID- 11480403 TI - Informed refusal. PMID- 11480404 TI - Operative vaginal delivery. Clinical management guidelines for obstetrician gynecologists. American College of Obstetrics and Gynecology. PMID- 11480405 TI - NHS database of reference costs is severely flawed. PMID- 11480406 TI - Approach to surgery in United Kingdom should be shaken up. PMID- 11480407 TI - Stakeholder health insurance. Commentary is disappointing. PMID- 11480408 TI - Stakeholder health insurance. Time for evidence based policy analysis. PMID- 11480409 TI - Warning to GMC. Patients should come first. PMID- 11480410 TI - Warning to GMC. Doctors are part of an economic hegemony. PMID- 11480411 TI - Rights involve responsibilities for patients. Doctors must not generally pass personal information about patients to others without consent. PMID- 11480412 TI - Rights involve responsibilities for patients. Research ethics committees will approve any protocol unless there are good reasons not to. PMID- 11480413 TI - Human papillomavirus testing. Authors' comments. PMID- 11480414 TI - Human papillomavirus testing. Effectiveness of testing for high risk HPV for triage of low grade abnormal smears is being assessed in TOMBOLA trial. PMID- 11480415 TI - The people's health assembly. PMID- 11480416 TI - Exercise prescription for older adults with osteoarthritis pain: consensus practice recommendations. A supplement to the AGS Clinical Practice Guidelines on the management of chronic pain in older adults. PMID- 11480417 TI - Possible mechanisms for tumour cell sensitivity to TNF-alpha and potential therapeutic applications. AB - TNF is a macrophage/monocyte-derived cytokine with cytostatic and cytotoxic anti tumour activity. TNF-alpha can cause haemorrhagic necrosis and regression of experimental tumours. Nevertheless, the TNF-alpha doses required to cure tumour bearing mice lead to injury of normal tissues and, eventually, may cause a lethal shock syndrome. This toxicity implies severe limitations for the therapeutic use of TNF-alpha. Reactive oxygen intermediates (ROls) are involved in TNF-alpha induced cell killing. Different studies are consistent with the hypothesis that tumour cell sensitivity to TNF-alpha is related to its capacity to buffer oxidative attack. Recently, we have demonstrated that the sensitivity of Ehrlich ascites tumor (EAT) cells to TNF depends on their glutathione (GSH, the most prevalent nonprotein thiol in mammalian cells) content and their rate of proliferation. This is important because tumour cell populations under active proliferative states may show higher GSH levels, and drug- and/or radiation resistant tumours have increased cellular levels of GSH. TNF-alpha induces a shift towards oxidation in the mitochondrial glutathione (mtGSH) status, a fact that is consistent with the hypothesis that mtGSH plays a key role in scavenging TNF-induced ROIs. GSH, which is not synthesized within mitochondria but is neccessary for their normal function, needs to be taken up from the cytosol through a high affinity multicomponent transport system. In consequence, different approaches that lead to depletion of mtGSH may improve the anticancer efficacy of TNF-alpha both in vitro and in vivo. As an example, EAT-bearing mice fed a glutamine-enriched diet (GED) show a selective increase of glutamate content witihin the tumour cells. Glutamate inhibits GSH uptake by tumour mitochondria and leads to a selective depletion of mtGSH content (not found in mitochondria of normal cells) to approx. 57% of the level found in tumour mitochondria of mice fed a standard diet (SD). Administration of rhTNF-alpha, which increases generation of mitochondrial ROIs, to EAT-bearing mice fed a SD does not affect significantly the rate of tumour growth. However, when tumour bearing mice fed a GED where treated with rhTNF-alpha the number of viable tumour cells was decreased to approx. 38% of controls. PMID- 11480418 TI - The importance of drug delivery to optimize the effects of bone morphogenetic proteins during periodontal regeneration. AB - Bone morphogenetic proteins (BMPs) include a large number of proteins belonging to the TGF-beta superfamily which are characterized by their ability to induce bone and cartilage formation. Since the isolation and purification of BMPs by recombinant technology, the effects of single BMPs can now be evaluated in animal models. Subcutanous placement of a single recombinant BMP, such as recombinant human (rh) BMP-2, in a rat ectopic assay shows recruitment of undifferentiated mesenchymal cells, cartilage formation, followed by replacement with bone, formation of its own bone marrow and physiological bone remodelling. The therapeutic use of recombinant BMPs in the treatment of periodontal disease (destruction of the tooth ligaments, surrounding bone and tooth cementum, the latter of which anchors the ligaments to the tooth surface from the adjacent tooth socket) has attracted considerable interest due to their potent ability to stimulate intramembranous bone formation without an endochondral intermediate. Their predictability in stimulating new bone may provide an alternative that has greater osteogenic potential than autogenous bone, other growth factors and bone substitutes. The biological processes and the potential role of growth factors involved in promoting regeneration are complicated by the involvement of different cell types each with their different growth rates and responses to various stimuli. The major cell types involved in periodontal regeneration include osteoblasts, cementoblasts and fibroblasts. Here, the formation of the new mineralized layers on the tooth and bone surfaces by cementoblasts and osteoblasts respectively are a prerequisite before periodontal ligament formation and attachment by fibroblasts can occur. In this regard, BMPs are likely candidates to stimulate periodontal regeneration because of their ability not only to promote osteogenesis but also to stimulate cementogenesis (new cementum formation). However, understanding when to manipulate each of the various cells differentiation pathway with the application of single or multiple doses of BMPs at the appropriate concentration is dependent upon a suitable delivery system that can be modified in order to optimize its effect during periodontal wound healing. Furthermore, treatment of intrabony periodontal defects with BMPs are likely to not only require appropriate temporal release of the agent, but also adaptation of a carrier that is robust enough to maintain its integrity around the coronal aspect of the root in order to provide space maintenance and support the mucoperiosteal flap. This review evaluates the effects of different delivery systems upon BMP-induced periodontal regeneration. PMID- 11480419 TI - Approaches to education of pharmaceutical biotechnology in faculties of pharmacy. AB - Pharmaceutical biotechnology is developing rapidly both in academic institutions and in the biopharmaceutical industry. For this reason, FIP Special Interest Group of Pharmaceutical Biotechnology decided to develop a questionnaire concerning pharmaceutical biotechnology education. After preliminary studies were completed, questionnaires were sent to the leading scientists in academia and research directors or senior managers of various Pharmaceutical Biotechnology Companies in order to gather their views about how to create a satisfactory program. The objectives of this study were as follows: -To review all of the graduate and undergraduate courses which are presently available worldwide on pharmaceutical biotechnology in Faculties of Pharmacy. -To review all of the text books, references and scientific sources available worldwide in the area of pharmaceutical biotechnology. When replying to the questionnaires, the respondents were asked to consider the present status of pharmaceutical biotechnology education in academia and future learning needs in collaboration with the biotechnology industry. The data from various pharmacy faculties and biotechnology industry representatives from Asia, Europe and America were evaluated and the outcome of the survey showed that educational efforts in training qualified staff in the rapidly growing field of pharmaceutical biotechnology is promising. Part of the results of this questionnaire study have already been presented at the 57th International Congress of FIP Vancouver, Canada in 1997. PMID- 11480420 TI - Biotechnology in the development of new vaccines and diagnostic reagents against tuberculosis. AB - Tuberculosis (TB) is a disease of global concern. About one third of the world population is infected with Mycobacterium tuberculosis. Every year, approximately 8 million people get the disease and 2 million die of TB. The currently available vaccine against TB is the attenuated strain of Mycobacterium bovis, Bacillus Calmette Guerin (BCG), which has failed to provide consistent protection in different parts of the world. The commonly used diagnostic reagent for TB is the purified protein derivative (PPD) of M. tuberculosis, which is nonspecific because of the presence of antigens crossreactive with BCG and environmental mycobacteria. Thus there is a need to identify M. tuberculosis antigens as candidates for new protective vaccines and specific diagnostic reagents against TB. By using the techniques of recombinant DNA, synthetic peptides, antigen specific antibodies and T cells etc., several major antigens of M. tuberculosis have been identified, e.g. heat shock protein (hsp)60, hsp70, Ag85, ESAT-6 and CFP10 etc. These antigens have shown promise as new candidate vaccines and/or diagnostic reagents against TB. In addition, recent comparisons of the genome sequence of M. tuberculosis with BCG and other mycobacteria have unraveled M. tuberculosis specific regions and genes. Expression and immunological evaluation of these regions and genes can potentially identify most of the antigens of M. tuberculosis important for developing new vaccines and specific diagnostic reagents against TB. Moreover, advances in identification of proper adjuvant and delivery systems can potentially overcome the problem of poor immunogenicity/short-lived immunity associated with protein and peptide based vaccines. In conclusion, the advances in biotechnology are contributing significantly in the process of developing new protective vaccines and diagnostic reagents against TB. PMID- 11480421 TI - Delivery of bioactive peptides and proteins across oral (buccal) mucosa. AB - The identification of an increasing array of highly potent, endogenous peptide and protein factors termed cytokines, that can be efficiently synthesized using recombinant DNA technology, offers exciting new approaches for drug therapy. However, the physico-chemical and biological properties of these agents impose limitations in formulation and development of optimum drug delivery systems as well as on the routes of delivery. Oral mucosa, including the lining of the cheek (buccal mucosa), floor of mouth and underside of tongue (sublingual mucosa) and gingival mucosa, has received much attention in the last decade because it offers excellent accessibility, is not easily traumatized and avoids degradation of proteins and peptides that occurs as a result of oral administration, gastrointestinal absorption and first-pass hepatic metabolism. Peptide absorption occurs across oral mucosa by passive diffusion and it is unlikely that there is a carrier-mediated transport mechanism. The principal pathway is probably via the intercellular route where the major permeability barrier is represented by organized array of neutral lipids in the superficial layers of the epithelium. The relative role of aqueous as opposed to the lipid pathway in drug transport is still under investigation; penetration is not necessarily enhanced by simply increasing lipophilicity, for other effects, such as charge and molecular size, also play an important role in absorption of peptide and protein drugs. Depending on the pharmacodynamics of the peptides, various oral mucosal delivery systems can be designed. Delivery of peptide/protein drugs by conventional means such as solutions has some limitations. The possibility of excluding a major part of drug from absorption by involuntary swallowing and the continuous dilution due to salivary flow limits a controlled release. However these limitations can be overcome by adhesive dosage forms such as gels, films, tablets, and patches. They can localize the formulation and improve the contact with the mucosal surface to improve absorption of peptides and proteins. Addition of absorption promoters/permeabilizers in bioadhesive dosage forms will be essential for a successful peptide/protein delivery system. PMID- 11480422 TI - The cellular and molecular basis of health benefits of grape seed proanthocyanidin extract. AB - Red grape seed extract containing proanthocyanidins and other antioxidants are being used as nutritional supplements by many health conscious individuals. The beneficial effects of grape seed proanthocyanidins (GSPE) have been reported, however, little is known about their mechanism(s) of action. One of the beneficial effects of GSPE is chemoprevention of cellular damage. The precise mechanism by which GSPE mediates, chemoprevention is not yet understood. This report addresses this issue. We investigated the mechanisms of actions of GSPE, which ameliorates chemotherapy-induced toxic effects of Idarubicin (Ida) and 4, hydroxyperoxycyclophosphamide (4-HC) in normal human Chang liver cells. Exposure to GSPE resulted in a significant reduction in apoptosis in response to the cytotoxicity of chemotherapeutic agents. RT-PCR analysis showed a significant increase in the anti-apoptotic gene Bcl-2 and a decrease in the cell cycle associated and proapoptotic genes, c-myc and p53 in cells treated with GSPE. These results suggest that some of the chemopreventive effects of GSPE are mediated by upregulating Bcl-2 and down regulating c-myc and p53 genes. PMID- 11480423 TI - Syntheses and effect of bombesin-fragment6-14 and its four analogues on food intake in rats. AB - A nonapeptide, H-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH2, corresponding to amino acids 6 to 14 of bombesin and its four analogues were synthesized by a solid phase method and were tested for their comparative effect on reducing activity on food intake using male Wistar rats. The synthetic bombesin-fragment6-14 showed reducing effect on food intake using male Wistar rats. Of the synthetic analogues, [Phe(4F)13]bombesin-fragment6-14 exhibited the most potent effect. The reducing effect on food intake of [Sar11]bombesin-fragment6-14 was lower than that of our synthetic bombesin-fragment6-14, but the other two analogues, [betaAla11]bombesin-fragment6-14 and [1-Nal8]bombesin-fragment6-14 showed no reducing effect on food intake in rats. PMID- 11480425 TI - Selegiline and mortality in subjects with Parkinson's disease: a longitudinal community study. PMID- 11480424 TI - Proposed diagnostic criteria for neurocysticercosis. AB - Neurocysticercosis is the most common helminthic infection of the CNS but its diagnosis remains difficult. Clinical manifestations are nonspecific, most neuroimaging findings are not pathognomonic, and some serologic tests have low sensitivity and specificity. The authors provide diagnostic criteria for neurocysticercosis based on objective clinical, imaging, immunologic, and epidemiologic data. These include four categories of criteria stratified on the basis of their diagnostic strength, including the following: 1) absolute- histologic demonstration of the parasite from biopsy of a brain or spinal cord lesion, cystic lesions showing the scolex on CT or MRI, and direct visualization of subretinal parasites by funduscopic examination; 2) major--lesions highly suggestive of neurocysticercosis on neuroimaging studies, positive serum enzyme linked immunoelectrotransfer blot for the detection of anticysticercal antibodies, resolution of intracranial cystic lesions after therapy with albendazole or praziquantel, and spontaneous resolution of small single enhancing lesions; 3) minor--lesions compatible with neurocysticercosis on neuroimaging studies, clinical manifestations suggestive of neurocysticercosis, positive CSF enzyme-linked immunosorbent assay for detection of anticysticercal antibodies or cysticercal antigens, and cysticercosis outside the CNS; and 4) epidemiologic- evidence of a household contact with Taenia solium infection, individuals coming from or living in an area where cysticercosis is endemic, and history of frequent travel to disease-endemic areas. Interpretation of these criteria permits two degrees of diagnostic certainty: 1) definitive diagnosis, in patients who have one absolute criterion or in those who have two major plus one minor and one epidemiologic criterion; and 2) probable diagnosis, in patients who have one major plus two minor criteria, in those who have one major plus one minor and one epidemiologic criterion, and in those who have three minor plus one epidemiologic criterion. PMID- 11480426 TI - Cognitive behaviour therapy for chronic fatigue syndrome. PMID- 11480427 TI - Cognitive behaviour therapy for chronic fatigue syndrome. PMID- 11480428 TI - Cognitive behaviour therapy for chronic fatigue syndrome. PMID- 11480429 TI - Cognitive behaviour therapy for chronic fatigue syndrome. PMID- 11480430 TI - Cognitive behaviour therapy for chronic fatigue syndrome. PMID- 11480431 TI - Cognitive behaviour therapy for chronic fatigue syndrome. PMID- 11480432 TI - Genetic testing and paternity. PMID- 11480433 TI - Fetal toxic effects and angiotensin-II-receptor antagonists. PMID- 11480434 TI - Testicular cancer and syndrome X. PMID- 11480435 TI - CD36 deficiency and insulin resistance. PMID- 11480436 TI - CD36 deficiency and insulin resistance. PMID- 11480437 TI - CD36 deficiency and insulin resistance. PMID- 11480438 TI - US pressure on less-developed countries. PMID- 11480439 TI - Attitudes towards smoking regulation in Italy. PMID- 11480440 TI - Screening with breast ultrasound in a population at moderate risk due to family history. AB - OBJECTIVE: To perform a pilot study to compare ultrasound and mammographic screening with particular respect to specificity, in a cohort of women at moderate family history risk of breast cancer. MATERIAL AND METHODS: High resolution ultrasound of both breasts was undertaken in 149 women at moderate risk of breast cancer who were referred for mammographic screening. The two tests were read blind to the other. Core biopsy was undertaken for solid lesions found on ultrasound. RESULTS: 149 Patients, mean age 42.15 years, were screened with bilateral ultrasound. 90 Examinations were normal. 46 Women had cysts, two had lesions previously biopsied and found to be benign, and one had a lymph node. 10 Focal solid lesions were biopsied on account of their ultrasound findings, 6.7% (95% confidence intervals (95% CIs) 2.7% to 10.7%). All but one of the 149 mammograms were normal. In this case both mammography and ultrasound showed a focal solid lesion that at core biopsy was found to be a fibroadenoma. Histology showed seven fibroadenomata, two areas of fibrocystic change, and one adenoid cystic carcinoma. Positive predictive value for biopsy was 10%. Mean follow up was 13.7 months. There was one interval cancer. CONCLUSION: Screening with breast ultrasound, in a cohort of women at moderate risk of breast cancer due to family history, has an acceptable biopsy rate. Screening with ultrasound and mammography in patients with an increased risk of breast cancer may be beneficial and a randomised study to examine issues of acceptability, reproducibility, and cost effectiveness is apt. PMID- 11480441 TI - Screening newborn infants for cystic fibrosis. PMID- 11480442 TI - The threat to the use of records and stored blood samples in medical screening research. PMID- 11480443 TI - Consenting adults. PMID- 11480444 TI - Precision of risk in multidimensional likelihood ratio based screening with first trimester screening for Down's syndrome as a special case. AB - OBJECTIVES: The development of computer programs for the calculation of risks and Monte Carlo estimation of the precision of such risks in likelihood ratio based screening with multiple gaussian distributed risk markers and a priori risks. A quantitative study of the variation of risk estimates in first trimester screening for Down's syndrome as a function of the variation of markers, and comparison of the results with published information on the variation of risk estimates in quality control schemes. METHODS: Algorithms for calculations in the multidimensional normal distribution and procedures for Monte Carlo simulation of risk distributions were implemented in the S-PLUS programming language and used to construct programs producing risk estimates and risk distributions. Parameters of risk marker distributions and correlations were obtained from the scientific literature. RESULTS: In screening for Down's syndrome during the first trimester the variation in risk estimates increased with increasing variation of biochemical and biometric markers, and the a posteriori risk may vary with at least a factor of 2-4. CONCLUSIONS: Risk estimates are not reasonable parameters in quality control systems. Instead, screening programmes should be controlled through careful monitoring of the distribution of risk estimates, in particular the screen positive rate, and control of the quality of the biochemical and biometric data. Furthermore, the correct classification of samples submitted for proficiency testing into screen positive and screen negative cases should be checked. PMID- 11480445 TI - Multiple marker second trimester serum screening for pre-eclampsia. AB - OBJECTIVE: To investigate whether an appropriate combination of maternal serum inhibin A, free beta-human chorionic gonadotropin (free beta-hCG), unconjugated estriol (uE3), and alpha-fetoprotein (AFP) may be an effective means of screening for pre-eclampsia in the second trimester of pregnancy. SETTING: Women who attended an antenatal clinic in Oxford, from whom serum samples were stored, 19 of whom subsequently developed pre-eclampsia. METHODS: Serum inhibin A, free beta hCG, uE3, and AFP were measured in 32 serum samples collected from the 19 women who developed pre-eclampsia and, for each sample, in three control samples collected from women with unaffected pregnancies matched for gestational age and maternal age. RESULTS: In pregnancies that developed pre-eclampsia the median inhibin A value was raised (1.7 multiples of the median (MoM) for unaffected pregnancies (95% confidence interval (95% CI) 1.1 to 2.7 MoM), the median free beta-hCG was raised (2.1, 1.4 to 3.3 MoM) and the median uE3 was lowered (0.8, 0.6 to 0.98 MoM) after 19 completed weeks of gestation and at least 2 weeks before the onset ofproteinuria. Values of AFP were similar in affected and unaffected pregnancies. Combining the values ofinhibin A, free beta-hCG, and uE3 to form a screening test would detect an estimated 55% of affected pregnancies with a false positive rate of 5%. CONCLUSIONS: Inhibin A, free beta-hCG, and uE3 in combination may be a useful screening test during the second trimester for pre eclampsia. PMID- 11480446 TI - Prevalence and family history of colorectal cancer: implications for screening. AB - OBJECTIVES: To estimate the current prevalence of family history and associated risk of prevalent colorectal cancer among middle aged and older British adults. METHODS: Cross sectional analysis of colorectal cancer and self reported family history based on data from a large population based study in Norfolk, United Kingdom. RESULTS: Of the 30,353 participants, 2,069 (6.8%) participants had reported a family history of colorectal cancer in at least one first degree relative. The prevalence of colorectal cancer in those with a family history was 1% and 0.5% in those without. Of the 151 participants with prevalent colorectal cancer, 14.6% reported a family history of the disease. Also, a family history of colorectal cancer in first degree relatives was significantly associated with an increased risk of prevalent colorectal cancer (multivariate odds ratio (OR) 2.32 (95% confidence interval (95% CI) 1.43 to 3.76). For those with one affected relative below the age of 45, and for those with two affected relatives the multivariate ORs were 5.29 (1.63 to 17.17) and 4.93 (1.17 to 20.70), respectively. CONCLUSIONS: Although positive family history may identify a group with increased likelihood of colorectal cancer, screening strategies targeted solely at people with a family history will have limited impact on reducing mortality from colorectal cancer at the population level. PMID- 11480447 TI - Estimating risk of breast cancer from population incidence affected by widespread mammographic screening. AB - OBJECTIVES: To estimate the absolute risk of breast cancer in women, allowing for the effect on incidence of the introduction of widespread mammographic screening. DESIGN: Annual breast cancer incidences were compared with numbers of annual mammograms in the population for 1980-96 to identify periods most likely to be affected by screening. Age specific breast cancer incidences 1972-96 were modelled by Poisson regression with an age, period, and cohort analysis. The 1996 age specific incidence was recalculated with the stable period effect 1972-89, and the age and cohort effects. Age specific incidence was converted to cumulative risk of breast cancer to age 79. SETTING: Population based data from all women in New South Wales (NSW), Australia. PATIENTS OR PARTICIPANTS: Breast cancer incidence in women 1972-96 obtained from the NSW Central Cancer Registry and female populations derived from successive censuses. Mammographic data from private sector mammograms (1985-96), and the mammographic screening service (1988 96) for NSW women. INTERVENTIONS: Introduction of population mammographic screening. MAIN OUTCOME MEASURES: Recorded age specific incidence and absolute risk of breast cancer to age 79 was compared with underlying incidence and cumulative absolute risk, adjusted for recent period effects, most likely due to mammographic screening in the population. RESULTS: The age, period, and cohort model showed an increasing effect for birth cohorts 1910-44 then a plateau, and prominent period effects in 1991 and 1994-6. Increased incidence of breast cancer coincided with an increase in mammographic examinations in the private sector (1991), and prevalent rounds of mammographic screening in the population (1994-6) after introduction of a statewide mammographic screening service. Recorded incidence produced a breast cancer risk to age 79 of 9.9% (1 in 10) for 1996, whereas estimation of underlying incidence yielded a risk of 8.5% (1 in 12). CONCLUSIONS: The introduction of mammographic screening in a population inflates the incidence of breast cancer because of diagnosis of prevalent cases. For the purpose of public and clinical communication, it is more reasonable and responsible to adjust for period effects (due to screening) rather than produce risk estimates based on recorded incidence, which may show an alarming increase in risk of breast cancer over a short period. PMID- 11480448 TI - Radiological peer review of interval cancers in the East Anglian breast screening programme: what are we missing? East Anglian Breast Screening Programme. AB - OBJECTIVE: To determine whether association between classification of interval cancers (false negative, minimal sign, true interval, occult) and tumour characteristics can indicate areas for improvement in effectiveness of screening. SETTING: East Anglian regional Breast: Screening Programme. METHOD: The consensus classification of interval cases, obtained through peer review, was compared with tumour characteristics: size; grade; and nodal status, and with radiological signs. A set of 923 cancers with known tumour characteristics, presenting in unscreened women, was available for comparison. RESULT: In total, 385 interval cases were analysed. Prognosis of interval cancers was at least as good as that of cancers in unscreened women, with fewer node positive cases. No significant relation was found between classification of cases and age of patients. True intervals tended to present later after a negative screen, although false negatives commonly presented earlier. Occult cancers were significantly smaller than false negatives. Analysis of histological type yielded little of relevance, and mammographic appearance was similar in all classifications. At diagnosis, cancers in the low risk group were more likely to be seen as spiculate masses, whereas high risk cases more often appeared as opacities. A higher proportion of interval cases had been assessed than in the normal screened population. CONCLUSION: Sensitivity may be improved by reducing false negative and minimal sign cases (21% of interval cancers in East Anglia). These will include tumours in all risk groups, and with all radiological features, thus no particular age group or radiological feature may be targeted for improvement. Misdiagnosis at assessment reduces sensitivity and therefore merits particular study. PMID- 11480449 TI - Detecting which invasive cancers at mammographic screening saves lives? AB - OBJECTIVES: Assessment of the features of primary operable breast carcinomas arising in women known to subsequently develop metastatic disease, to show for which invasive breast cancers earlier detection by mammographic screening is potentially beneficial. These data were applied to a separate series of screen detected cancers. METHODS: Features associated with the development of metastatic disease after a previous operable breast cancer were ascertained from examination of histological sections of the initial primary carcinoma and in particular the incidence of nodal positivity and definite vascular invasion. Trends in the frequency of nodal involvement and vascular invasion according to histological grade, invasive size, and tumour type were then examined in a further group of 573 screen detected invasive cancers to predict the likelihood of development of systemic disease in these women. RESULTS: Of 173 women who developed metastatic disease after a previous operable breast cancer, 79 (72%) had nodal metastases and 62 (59%) had definite vascular invasion. A high proportion (84%) had either lymph node metastases or vascular invasion or both. The absence of vascular invasion and nodal involvement in invasive breast cancer indicated a low risk of subsequent development of metastatic disease. In the screen detected group, grade 1 invasive cancers <20 mm in size and grade 2 and 3 cancers <10 mm in size had low rates of nodal involvement and vascular invasion. There was a gradual trend to small size, lymph node negativity, and less vascular invasion when comparing screen detected ductal carcinoma of no special type, tubular mixed carcinoma, and tubular carcinoma. Cancers with a lobular component tended to be larger and more often lymph node positive than ductal and no specific type carcinomas. CONCLUSIONS: These data suggest that identification of grade 1 cancers less than 20 mm in size and grade 2 and 3 cancers less than 10 mm in size at screening is likely to be beneficial, with a lower likelihood of developing metastatic disease from these lesions. The detection of tubular mixed carcinomas and ductal carcinomas of no specific type also appears beneficial. However the detection of tumours with lobular features at breast screening does not seem to significantly benefit the patient. PMID- 11480450 TI - Cluster randomised controlled trial comparing the effectiveness and cost effectiveness of two primary care interventions aimed at improving attendance for breast screening. AB - OBJECTIVES: To examine the effectiveness and cost-effectiveness of two interventions based in primary care aimed at increasing uptake of breast screening. SETTING: 24 General practices with low uptake in the second round of screening (below 60%) in north west London and the West Midlands, UK. Participants were all women registered with these practices and eligible for screening in the third round. METHODS: Pragmatic factorial cluster randomised controlled trial, with practices randomised to a systematic intervention (general practitioner letter), an opportunistic intervention (flag in women's notes prompting discussion by health professionals), neither intervention, or both. Outcome measures were attendance for screening 6 months after the practices had been screened and cost-effectiveness of the interventions. RESULTS: 6,133 Women were included: 1,721 control; 1,818 letter; 1,232 flag; 1,362 both interventions. Attendance data were obtained for 5,732 (93%) women. The two interventions independently increased breast screening uptake in a logistic regression model adjusted for clustering, with the flag (odds ratio (OR) 1.43, 95% confidence interval (95% CI) 1.14 to 1.79; p=0.0019) marginally more effective than the letter (OR 1.31, 95% CI 1.05 to 1.64; p=0.015). Health service costs per additional attendance were 26 pounds (letter) and 41 pounds (flag). CONCLUSIONS: Although both interventions increased attendance for breast screening, the letter was the more cost-effective. Any decision to implement both interventions rather than just the letter will depend on whether the additional (41 pounds) costs are judged worthwhile in terms of the gains in breast screening uptake. PMID- 11480451 TI - Improving attendance for breast screening among recent non-attenders: a randomised controlled trial of two interventions in primary care. AB - OBJECTIVES: To examine the effectiveness and cost-effectiveness of two primary care based interventions aimed at increasing breast screening uptake for women who had recently failed to attend. SETTING: 13 General practices with low uptake in the second round of breast screening (below 60%) in north west London and the West Midlands, United Kingdom. Participants were women in these practices who were recent non-attenders for breast screening in the third round. METHODS: Pragmatic factorial randomised controlled trial, with people randomised to a systematic intervention (general practitioner letter), an opportunistic intervention (flag in women's notes prompting discussion by health professionals), neither intervention, or both. Outcome measures were attendance for screening 6 months after randomisation and cost-effectiveness of the interventions. RESULTS: 1,158 Women were individually randomised as follows: 289 control; 291 letter; 290 flag; 288 both interventions. Attendance was ascertained for 1,148 (99%) of the 1,158 women. Logistic regression adjusting for the other intervention and practice produced an odds ratio (OR) for attendance of 1.51 (95% confidence interval (95% CI 1.02 to 2.26; p=0.04) for the letter, and 1.39 (95% CI 0.93 to 2.07; p=0.10) for the flag. Health service costs/ additional attendance were 35 pounds (letter) and 65 pounds (flag). CONCLUSIONS: Among recent non-attenders, the letter was effective in increasing breast screening attendance. The flag was of equivocal effectiveness and was considerably less cost-effective than the letter. PMID- 11480452 TI - Oxidized low density lipoprotein: the occurrence and metabolism in circulation and in foam cells. AB - Oxidatively modified low density lipoprotein (OxLDL) is thought to be involved in the early development of atherosclerotic lesions. The appearance of lipid-laden foam cells is known to be one of the typical features of atherosclerotic lesions, and accumulating evidence has demonstrated that foam cells are formed after taking up OxLDL by macrophages in vitro. However, the modified structures, distribution, and metabolism of OxLDL present in vivo are poorly understood. Recently, our studies, together with others, have demonstrated that OxLDL is actually present in circulating human plasma. Furthermore, we have provided evidence that foam cells accumulate modified apoB fragments derived from OxLDL in the cells. This article reviews recent progress in this field, including the intracellular metabolism of OxLDL in foam cells and the relevance of OxLDL as an in vivo ligand for macrophages. PMID- 11480453 TI - A comparative study of cultured smooth muscle cell proliferation and injury, utilizing glycated low density lipoproteins with slight oxidation, auto oxidation, or extensive oxidation. AB - We investigated the influence of glycated low density lipoprotein (LDL) for vascular smooth muscle cell (SMC) proliferation or injury. We utilized glycated, slightly oxidized LDL (GLDL-LOX), glycated, auto-oxidized LDL (GLDL) and glycated, metal-induced extensively oxidized LDL (GLDL-OX) to examine the effect of glycation itself or combined glycation and oxidation on SMC. GLDL-LOX induced SMC proliferation and migration, and increased the number of platelet-derived growth factor receptor, beta subunits, (PDGF-R) positive SMC. Also, GLDL-LOX promoted protease activity, compared with the other groups including native LDL (control). GLDL and GLDL-OX demonstrated SMC injury with apoptosis and Bax protein expression, compared with native LDL and GLDL-LOX. These results suggested that LDL glycation contributed to the progression of atherosclerosis by promoting SMC migration and proliferation, with little dependence on oxidative modification. Secondary auto-oxidation adding to glycation induced SMC apoptosis, and SMC injury occurred in the state of strong oxidation with glycation. We concluded that LDL glycation might play a key role in the progression of atherosclerosis in diabetes, and glycated LDL promoted atherosclerosis, even with little assistance from oxidation. PMID- 11480454 TI - Relative induction of mRNA for HMG CoA reductase and LDL receptor by five different HMG-CoA reductase inhibitors in cultured human cells. AB - The effect of various 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors on the induction of HMG-CoA reductase and low density lipoprotein (LDL) receptor mRNA were quantitatively determined in the cultured human hepatoma cell line Hep G2 by means of a ribonuclease protection assay. Lipophilic inhibitors including mevastatin, simvastatin, atorvastatin and NK-104 were able to increase the levels of mRNAs for HMG-CoA reductase and the LDL receptor, but the hydrophilic inhibitor pravastatin was not effective in Hep G2 cells as had previously been reported. The LDL receptor mRNA was induced by NK-104 most effectively between 0.1 to 10 microM among the lipophilic inhibitors, whereas the degrees of induction of HMG-CoA reductase mRNA by these inhibitors did not differ significantly from each other. When cells were treated with a 200-fold excess of the IC50 concentration of each inhibitor, NK-104 was able to induce LDL receptor mRNA most effectively. These results indicate that the effect of HMG-CoA reductase inhibitors on the upregulation of mRNA for reductase and LDL receptor are different from each other and among these lipophilic inhibitors. NK-104 is most effective in inducing LDL receptor mRNA in Hep G2 cells. PMID- 11480455 TI - Genomic structure and mapping of human orphan receptor LXR alpha: upregulation of LXRa mRNA during monocyte to macrophage differentiation. AB - In order to identify changes in the gene expression profile during human monocyte/macrophage differentiation in the presence of GM-CSF, the expression level of various mRNA was studied using DNA microarray technology. We found LXR alpha (LXRa) to be the most highly induced transcriptional regulator during macrophage differentiation. The LXRa mRNA level was induced 40 fold which ranked it as the 10th highest among the approximately 5,600 genes studied. Although only restricted hepatic expression of LXRa mRNA had been reported, the macrophage expressed the highest level of LXRa among the nine human tissues and cultured cells studied. To further investigate transcriptional control, we have characterized the genomic structure of the human LXRa gene and determined the structure of its promoter region. The human LXRa gene consists of eleven exons, and analysis of the promoter region indicated the presence of conserved binding sites for myeloid zinc finger protein 1, which may be related to the extrahepatic expression of LXRa. LXRa is known to be activated by oxysterols, and the induced expression of the gene may be related to the foam cell formation in atherosclerotic lesions. PMID- 11480456 TI - Serum paraoxonase and arylesterase activities in hemodialysis patients. AB - Cardiac death from atherosclerosis is common in hemodialysis patients. Human serum paraoxonase (PON1), an esterase, is associated with high-density lipoprotein and inhibits the susceptibility to oxidization of low-density lipoprotein (LDL). The PON1 genetic polymorphisms of 192 Gln/Arg and 55 Leu/Met in the amino acid sequence are partly involved in the PON1 enzyme activity. We investigated the PON1 enzyme activities for paraoxon (paraoxonase) and phenylacetate (arylesterase), and the two polymorphisms in 96 patients undergoing hemodialysis and in 136 normal controls. Both activities were significantly lower in the hemodialysis patients than in the controls (97+/-43 vs 155+/-57 micromol/min/l for paraoxonase, and 71+/-20 vs 92+/-22 mmol/min/l for arylesterase, respectively). There was no difference in the distribution of the two polymorphisms between patients and controls, and in every subgroup classified by the polymorphisms, both paraoxonase and arylesterase activities were lower in patients than in controls. This suggested that the enzyme activities of PON1 decreased in hemodialysis patients, independent of the genetic polymorphism. The decrease in PON1 enzyme activity in hemodialysis patients may modify a susceptibility to oxidization of LDL, which contributes to an acceleration of atherosclerosis. PMID- 11480457 TI - Troglitazone improves endothelial dysfunction in patients with insulin resistance. AB - Insulin resistance is a possible major metabolic cause of atherosclerosis. Endothelial dysfunction is commonly found in patients with insulin resistance, and primary treatment of insulin resistance with troglitazone should improve such endothelial dysfunction. Thus, the effects of troglitazone on endothelial function were investigated. Thirteen non-diabetic male subjects with hyperinsulinemic response to oral glucose load (n = 7) and normal (n = 6) subjects were investigated. Flow-mediated dilatation (FMD) of the brachial artery was examined by high resolution ultrasonography before and after the administration of troglitazone of 400 mg for 4 weeks. In insulin resistant subjects, fasting glucose (4.9+/-0.3 to 4.7+/-0.3 mmol/L, p<0.05), insulin (45+/ 30 to 25+/-15 pmol/L, p<0.05) and response to oral glucose load (AUC glucose: 15.0+/-3.5 to 13.0+/-2.2 mmol x h/L, p<0.05; AUC insulin: 965+/-560 to 475+/-275 pmol x h/L, p<0.05) were significantly reduced. FMD was significantly improved in insulin resistant subjects. A significant negative correlation was observed between FMD and AUC insulin (r=-0.64, p<0.05). The present study demonstrates that FMD is impaired in insulin resistant subjects, and troglitazone improves the blunted vascular response and impaired insulin response. This finding suggests that primary treatment of insulin resistance could prevent the development of atherosclerosis by improving endothelial dysfunction. PMID- 11480458 TI - Effects of cacao liquor polyphenols on the susceptibility of low-density lipoprotein to oxidation in hypercholesterolemic rabbits. AB - The effects of cacao liquor polyphenols (CLP) on the susceptibility of low density lipoprotein (LDL) to oxidation in hypercholesterolemic rabbits were examined. Six Japanese white rabbits which had been fed a high cholesterol diet (HCD) for 3 weeks were fed HCD containing 1% CLP for the following 10 days. The susceptibility of LDL to oxidation induced by 2-2'-azobis(4-methoxy-2, 4 dimethylvaleronitrile) (V-70) was evaluated by measuring the production of conjugated dienes and thiobarbituric acid reactive substances (TBARS). The lag time was significantly prolonged from 37.7 min before intake of CLP to 42.9, 44.2 and 45.8 min after 4, 7 and 10 days of CLP intake. TBARS production after intake of CLP was also markedly reduced compared with the level before intake. There was no difference in plasma lipid concentrations comparing the levels before and after CLP intake. In conclusion, in hypercholesterolemic rabbits, orally administered CLP was absorbed and distributed to the blood, and the resistance of LDL to oxidation was thereby increased. PMID- 11480459 TI - Effects of advanced glycation end products on the proliferation and fibronectin production of smooth muscle cells. AB - The aim of this study was to evaluate the effects of advanced glycation end products (AGEs) on the proliferative activity and fibronectin production of smooth muscle cells (SMCs). AGE-bovine serum albumin (AGE-BSA) was prepared by incubation with D-glucose at 37 degrees C for 60 days. Cultured SMCs were obtained from explants isolated from porcine abdominal aorta and used between passages 3 and 10. The proliferative activity of SMCs was examined by MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and by incorporation of 3H-thymidine into DNA. Fibronectin production was assessed by competitive ELISA assay for both fibronectin secreted into the culture medium (M FN) and cell-associated fibronectin (C-FN), i.e., both intra- and peri-cellular fibronectin. Theassay revealed that AGE-BSA did not produce any change in optical density (A570) of SMCs at concentrations of up to 20 microg/ml, but decreased that of SMCs at a concentration of 40 microg/ml. The addition of PDGF (5 ng/ml) induced an increase in 3H-thymidine incorporation into DNA of quiescent SMCs, while the addition of AGE-BSA (20 microg/ml) had no effect. In contrast, AGE-BSA significantly increased C-FN of SMCs (30.8+/-8.58 ng/microg TP), compared to unmodified BSA (16.5+/-4.19 ng/microg TP). However, no difference in M-FN levels was observed between cells treated with AGE-BSA and unmodified BSA. The addition of anti-transforming growth factor (TGF)-beta antibody restored the levels of C FN in SMCs cultured in 20 microg/ml of AGE-BSA, suggesting that TGF-beta might act as an intermediate factor in AGE-induced fibronectin production by SMCs. Our results suggest that interaction of AGE-modified proteins with SMCs may play a role in the development of atherosclerosis in diabetic and non-diabetic patients. PMID- 11480460 TI - The volume limit in fluid resuscitation to prevent respiratory failure in massively burned children without inhalation injury. AB - We evaluated the accurate fluid requirement to prevent respiratory failure during the postresuscitation period in the resuscitation of massively burned children without inhalation injury. Forty-nine children were treated by similar fluid resuscitation and physiologic support protocols. Using a retrospective chart review, the children were divided into three groups as follows: Group N (no lung injury, n = 33, 41.4+/-18.7%TBSA burned), Group M (mild-to-moderate lung injury, n = 11, 73.7+/-17.1%TBSA burned) and Group S (severe lung injury, n = 5, 67.2+/ 16.6%TBSA burned). Information about fluid resuscitation during the first 24 hr post-injury was collected and compared among the three groups. There was no significant difference in the hourly urine output and the resuscitation volume estimated by body weight and burn size among the groups. The volumes estimated by ml/kg/%TBSA burned were 7.0 ml/kg/%TBSA burned, 8.0 ml/kg/%TBSA burned, and 9.4 ml/kg/%TBSA burned in Groups N, M, and S, respectively. According to the fluid volume estimated by the burn index (BI; 1/2 of % second-degree burns plus % third degree burns), the volumes were 13.8+/-4.0 ml/kg/BI, 14.4+/-4.4 ml/kg/BI, 18.8+/ 3.7ml/kg/BI in Groups N, M, and S, respectively (Group N < Group S, p < 0.05). There was a significant positive correlation between the maximum respiratory index (AaDO2/PaO2) during the first week and the initial total volume administered (ml/kg/BI). These findings indicated that the fluid requirements to prevent postresuscitation respiratory failure in massively burned children might be estimated according to the depth of burned area in addition to body weight and burn size. PMID- 11480461 TI - A prospective multicenter trial to determine the incidence of transient neurologic symptoms after spinal anesthesia with phenylephrine added to 0.5% tetracaine. AB - The addition of vasoconstrictors for spinal anesthesia is controversial, since an increase in the incidence of transient neurologic symptoms (TNS) has been reported. A multicenter, randomized, double-blind study was conducted to assess the effectiveness of spinal anesthesia with phenylephrine in addition to tetracaine as well as the incidence of neurological complications. We studied 64 patients with comparable demographic characteristics who were scheduled for elective surgery for a lower limb, or a gynecological or urological procedure. The patients were allocated randomly into 2 groups. Group P (n = 34) received 0.5% tetracaine in 10% glucose with 0.025% phenylephrine, while group C (n = 30) received 0.5% tetracaine in 10% glucose. Our results showed that only 2 patients (6.7%) in group C experienced TNS, and their symptoms disappeared within 72 hr after anesthesia, while none of the patients (0%) in group P complained of symptoms. The incidence of TNS was thus not significantly different between the two groups. Six hours after the sensory block, group P patients demonstrated sensory disturbance, with the median spinal dermatome corresponding to the L1 segment. Moreover, systolic blood pressure in group P was significantly higher than that in group C, 5 min, 15 min, and 20 min after injection. The incidence of TNS in the present study does not seem to be greater after surgery with spinal anesthesia using 0.5% hyperbaric tetracaine and 0.5 mg phenylephrine than without phenylephrine. Randomized, double-blind, cross-over trials with a larger sample size would be required in the future to obtain more reliable results. PMID- 11480462 TI - A morphometric study on postnatal development of the external granular layer of mice cerebella, focusing on local difference. AB - The external granular layer (EGL) of the cerebellum thickens, thins and disappears in its developmental process. We examined the thickness of the EGL, both intralobule differences and interlobule differences, in the whole midline sections of mouse cerebella for the entire postnatal period up to disappearance. The thinnest site in each lobule was located at the outer apex throughout the observation period, the thickest site was the portion facing the inner apex during early period, and that facing the convexity where the EGL curved in the later period. The observed interlobular differences of the EGL thickness were statistically divided into three groups, referred to as the early developing group (EDG), the late developing group (LDG), and the mixed-type group (MTG). The EDG consisted of the whole anterior lobe and a site in lobule VI facing the fissura prima. These sites thickened earlier, and showed a similar thickness on each observation day. The LDG was composed of all sites in the posterior lobe, with the exception of two sites where lobules VIII and IX confronted each other as well as the site included in the EDG. The sites in the LDG thickened later. They demonstrated a similar thickness during thickening, but varied during thinning. The MTG, consisting of two sites where lobules VIII and IX facied each other, showed features similar to the EDG in the mitotic zone and the LDG in the premigratry zone. These data may serve as the basis for studies on regional differentiation of the cerebellum. PMID- 11480463 TI - Influence of initial status on functional gain for Japanese patients with first cerebral hemorrhage. AB - It is important to identify in advance patients who will achieve the greatest functional gains from rehabilitation therapy, as specialist rehabilitation resources are still scarce in Japan. The purpose of this study was to determine whether functional score at admission influences the functional change (functional score at discharge minus functional score at admission) after inpatient rehabilitation for first cerebral hemorrhage. One hundred and ninety three patients with cerebral hemorrhage were enrolled in this study. They were assessed using the Functional Independence Measure (FIM) at admission and discharge and underwent inpatient rehabilitation treatment. Patients were stratified into 3 groups according to their FIM total scores on admission as follows: (1) < or = 36 (severely affected patient group); (2) 37-72 (moderately affected patient group); and (3) >73 (mildly affected patient group). Scheffe's multiple comparison test showed that patients in group 1 were significantly older (mean +/- SD = 63 +/- 10 years) than those in groups 2 (56 +/- 10 years) or 3 (53 +/- 12 years). Patients in group 2 showed significantly greater FIM gain (37 +/- 17) compared with patients in groups 3 (23 +/- 12) or 1 (27 +/- 23). The results suggest that moderately affected patients at admission will show significantly higher functional gain compared with severely or mildly affected patients. Mildly affected patients at admission had a significantly shorter length of hospital stay for rehabilitation than the other groups. There was no significant difference in onset to admission interval between the 3 groups. The functional levels of affected patients on admission, as stratified by the FIM scale, roughly predict the degree of functional gain following rehabilitation in patients with first cerebral hemorrhage. Moderately affected patients will benefit from intensive rehabilitation. This study may be useful in determining how best to prioritize rehabilitation therapy. PMID- 11480464 TI - Central and peripheral haemodynamics in individuals with paraplegia during light and heavy exercise. AB - The purpose of this study was to analyse lower leg skin blood flow (laser Doppler flowmetry) in five individuals with high-level paraplegia (T5-T9), six participants with low-level paraplegia (T10-T12) and six able-bodied controls during 3-minute light and heavy arm-cranking exercises (approximately 15% and 80% peak power output, respectively). Throughout light exercise, cutaneous vasoconstriction was shown for the control group (-20%), but not the low-level (+62%) or the high-level paraplegic group (+33%). During heavy exercise, vasoconstriction was initially found for controls followed by an increase in skin blood flow during the last 2 minutes, whereas the participants with paraplegia demonstrated skin blood flow increases. Skin blood flow responses were not related to lesion level. Metabolic parameters were not different among the three groups, but heart rates for participants with paraplegia were higher during heavy exercise than in controls. These results suggest impaired sympathetic vasoconstriction in individuals with paraplegia during exercise. PMID- 11480465 TI - Localized low back pain and low back pain as part of widespread musculoskeletal pain: two different disorders? A cross-sectional population study. AB - In a cross-sectional postal questionnaire study we compared individuals with localized low back pain (LBP) with individuals with LBP as part of widespread musculoskeletal pain, according to demographic and lifestyle characteristics and functional ability. All the inhabitants in Ullensaker county born 1918-20, 1928 30, 1938-40, 1948-50, 1958-60 and 1968-70 were sent a questionnaire in 1994. The study population comprised 2,893 responders. LBP as part of widespread pain indicated reduced functional ability, and the groups differed in several demographic and lifestyle characteristics. PMID- 11480466 TI - "Active back school", prophylactic management for low back pain: three-year follow-up of a randomized, controlled trial. AB - The purpose of the present study was to investigate the long-term effect of the Active Back School programme on minimizing recurrences of episodes of low back pain. Forty-three subjects were randomly allocated to the Active Back School group and 38 to the control group. There were no significant differences between the groups with regard to baseline characteristics. The Active Back School programme comprised 20 lessons each divided into a 20-min theoretical and a 40 min exercise part during a 13-week period. Nine participants (11%) dropped out during the study period. Recurrence of new low back pain episodes was significantly less (p = 0.04), and the time from inclusion to the first new low back pain episode was significantly on the side of the Active Back School group (p < 0.01). The duration of sick leave was found to be significantly shorter (p < 0.01) in the Active Back School group compared to the control group. The Active Back School reduced the recurrence and severity of new low back pain episodes at 36 months' follow-up. PMID- 11480467 TI - Physical measurements and questionnaires as diagnostic tools in chronic low back pain. AB - The objective of this study was to assess the diagnostic value of common questionnaires and measures of physical performance in low back pain (LBP) syndrome. One hundred and fourteen patients with LBP classified according to the Quebec Task Force were compared with 50 patients with different pain syndromes but without apparent LBP. The discriminating value of each variable was estimated by calculating the area under the receiver operating characteristics (ROC) curve. The diagnostic value of the Million and Oswestry disability questionnaires was evident, with the area under the ROC curve varying between 0.73 and 0.88. The isometric trunk extension-flexion strength test with concomitant reaction-time test could not distinguish between patients (area under ROC curve 0.50-0.68). Sensitivity of pain drawing was excellent but specificity was low: 47% for men and 39% for women. In conclusion, disability questionnaires have discriminating power. The trunk muscle strength test does not perform well as a diagnostic tool. The area under the ROC curve and the use of other patients as controls make it easier to assess the diagnostic specificity of a particular method. PMID- 11480468 TI - Intra- and inter-tester reliability and reference values for hand strength. AB - The intra- and inter-tester reliability for measurement of handgrip strength and indexgrip strength using the Jamar dynamometer was investigated in 32 healthy volunteers, and the intra-tester reliability in 13 patients with cervical radiculopathy. The results from the reliability studies showed that handgrip and indexgrip strength measured with the Jamar dynamometer is a reliable method (ICC values 0.85-0.98) and can be recommended for use in clinical practice. Age- and sex-specific reference values for handgrip strength and indexgrip strength were measured with the Jamar dynamometer in 101 randomly selected healthy volunteers, aged 25-64 years. The results from the reference value study showed that sex is a more important determinant of hand strength than age, height and body weight. The reference values for hand strength improve the potential for objective evaluation of patients with arm/hand disorders caused by cervical radiculopathy. PMID- 11480469 TI - Ability and perceived difficulty in daily activities in people with poliomyelitis sequelae. AB - Polio involvement and its relation to ability in terms of independence and perceived difficulty in activities in daily living (ADL) were studied in outpatients (median age 57 years) with poliomyelitis sequelae (n = 133). The use and need of assistive devices and housing conditions were recorded. The subjects had a high degree of independence in personal ADL, whereas more than 50% were dependent on cleaning, shopping and transportation. A considerable number of subjects independent in ADL reported difficulties in daily activities. The majority of interventions in the form of housing adaptation and assistive devices were provided for persons severely weakened by polio. Ratings of perceived difficulty indicated need of interventions to prevent overload and increased dependence in ADL. However, about half of the recommended mobility assistive devices were rejected. The results indicate that polio subjects try to maintain their independence despite perceived difficulties and may be reluctant to use assistive devices. PMID- 11480470 TI - Low effectiveness of prescribed partial weight bearing. Continuous recording of vertical loads using a new pressure-sensitive insole. AB - To enable objective measurements of weight bearing in hip replacement patients a portable instrument set-up with an on-line registering pressure-sensitive insole was developed. Six men and 9 women, median age 58 (48-67) years, who had been operated on with a cementless or hybrid hip arthroplasty were studied. All patients were independent and functional. A physiotherapist instructed the patients to use crutches in order to support 30% of their body weight. The patients then walked a standardized distance with crutches over five different types of terrain: level, uphill, downhill, upstairs and downstairs. The trial was repeated once. In both men and women most of the steps taken resulted in a load of >30% of body weight. None of the patients managed to comply with the directive, even though five of them thought they did. The type of terrain had no significant influence on the pattern of load. Our findings indicate that the effectiveness of prescribing limited weight bearing is questionable. PMID- 11480471 TI - Guidelines to statistical evaluation of data from rating scales and questionnaires. PMID- 11480472 TI - Current guidelines for the management of unstable angina: a new diagnostic and management paradigm. AB - The new guidelines reflect a worldwide change to more sensitive diagnostic strategies and more aggressive management of unstable angina. Stratification to high risk now includes those patients with only a minor degree of electrocardiographic ST depression (0.5 mm) or a significant elevation of cardiac troponin. High-risk patients are recommended to be treated with intensive medical and invasive management. Intermediate-risk patients may be best evaluated using an accelerated diagnostic strategy over an 8-12 h period before being reclassified as high or low risk. PMID- 11480473 TI - Lung volume reduction surgery: the Thoracic Society of Australia and New Zealand. AB - Lung volume reduction surgery involves the removal of emphysematous lung tissue with the aim of palliating symptoms in selected patients with severe emphysema. This form of surgery is being practised in Australia with favourable short-term outcomes, similar to those reported in the literature. Large multicentre trials are currently underway in North America and the United Kingdom to clarify issues of safety and long-term efficacy. As a result, it is too early to apply an evidence-based approach to this procedure. In the meantime, local audits of practice need to be undertaken to define patient subgroups at higher risk of morbidity and mortality. PMID- 11480474 TI - Angiotensin-1-converting enzyme and angiotensinogen gene polymorphisms in Maori and Pacific Island people in New Zealand. PMID- 11480475 TI - What do Australians want from medicine? PMID- 11480476 TI - Medical education in 2001: the place of the medical humanities. PMID- 11480477 TI - Insulinoma masked by pregnancy. PMID- 11480478 TI - Reactivation of latent melioidosis in association with staphylococcal endocarditis. PMID- 11480479 TI - First case of Nipah virus encephalitis in Singapore. PMID- 11480480 TI - Guidelines for acute coronary syndromes: translating the evidence to best practice. PMID- 11480481 TI - Monoclonal antibody therapy for Crohn's disease. PMID- 11480482 TI - Why complaining is good for medicine. PMID- 11480483 TI - Women's health after plastic surgery. AB - BACKGROUND: Allegations that exposure to endogenous silicone, especially related to breast implants, might be causally related to connective tissue disease originated from case studies. More recent comparative studies have implied no such increased risk. The aims of the present study were to compare the prevalence and/or incidence of autoimmune and connective tissue disorders in a population based cohort of female Sydney residents stratified by augmentation mammoplasty status. METHODS: In this population-based retrospective cohort study, the health status of female Sydney residents who had augmentation mammoplasty for cosmetic reasons between 1979 and 1983 was compared with that of female Sydney residents who had non-silicone-associated plastic surgery over the same period. Both groups were matched for age (+/- 5 years), year of plastic surgery (+/- 2 years), plastic surgeon, anaesthetist and mode of anaesthesia. Outcome measures comprised rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, sicca symptoms polymyositis/ dermatomyositis, connective tissue disease overlap, digital vasospasm, abnormal nailfold capillaroscopy, elevated antinuclear antibody titre, carpal tunnel syndrome, tendonitis, osteoarthritis, psoriatic arthritis, livedo reticularis, thyroid disease, multiple sclerosis, axillary lymphadenopathy, fibromyalgia and breast carcinoma. RESULTS: There was no difference in the occurrence of connective tissue diseases or connective tissue disease-related parameters, thyroid disorders, fibromyalgia or multiple sclerosis between cohorts. However, axillary adenopathy and low titre positive antinuclear antibody (ANA) occurred with a significantly greater frequency in the exposed cohort (odds ratio (OR) = 3.50, 95% confidence interval (CI) = 2.10-5.84 and OR = 1.29, 95% CI = 1.03-1.62, respectively). Axillary adenopathy correlated with capsular contracture (relative risk (RR) = 2.07, 95% CI = 1.22-3.51) and also the self-reported development of digital vasospasm (RR = 3.20, 95% CI = 1.46-7.03) after breast augmentation. CONCLUSIONS: No association was found between augmentation mammoplasty exposure and various connective tissue diseases and/or their related features. However, axillary adenopathy and low titre ANA were detected more frequently in the exposed cohort. Women with axillary adenopathy were more likely to have breast capsular contracture and report digital vasospasm post-dating surgery. Given comparable frequencies of higher titre ANA of both cohorts, the finding of elevations of low titre ANA is of dubious clinical significance. PMID- 11480484 TI - Changes in hepatitis C-related liver disease in a large clinic population. AB - BACKGROUND: Hepatitis C virus (HCV) infection is a significant problem in the Australian community. Over the past few years, the number of patients with diagnosed hepatitis C has increased greatly. The aims of the present study were to define the clinical features of a large group of patients with chronic HCV infection and to examine changes occurring in the referral base and epidemiological characteristics of this group since analysis of the first 342 patients in 1994. METHODS: The study included 1,546 consecutive anti-HCV-positive patients who had been referred to St Vincent's Hospital from January 1990 to June 1998. Clinical and laboratory data were collected on all patients. RESULTS: Referrals from general practitioners increased from 31% to 70% of all patients between 1990-1993 and 1994-1998. A history of injecting drug use (IDU) was present in 64% of the patients. While 89% of the IDU group was Australasian born, 49% of those in the sporadic group were born overseas. Cirrhosis was found in 18% of biopsied patients. Age, infection duration, age at infection, Mediterranean or Asian origin and a history of transfusion or lack of HCV risk factors were associated with cirrhosis on univariate analysis. Patient age was the only independent predictor of cirrhosis. CONCLUSION: The majority of patients with HCV are diagnosed in general practice. A risk factor for infection is identified in 82% of patients. While our reported prevalence of cirrhosis may be an overestimate of that in the overall HCV community, the ultimate disease burden is likely to be significant. PMID- 11480485 TI - Prospective study of 424 cases of Staphylococcus aureus bacteraemia: determination of factors affecting incidence and mortality. AB - BACKGROUND: Staphylococcus aureus bacteraemia (SAB) is a common complication of S. aureus infection and is associated with a high mortality. AIMS: To document prospectively the pattern of illness associated with SAB in New Zealand and, by recording patient demographic factors and clinical features, to identify risk factors associated with a poor outcome. METHODS: From 1 July 1996 to 31 December 1997, adults with SAB were prospectively studied in six tertiary care hospitals. All information obtained from patients' records was recorded on worksheets and transferred to a computerized spreadsheet for analysis. RESULTS: There were 424 patients with SAB. Maori (relative risk (RR)= 1.8, 95% confidence interval (CI) = 1.3-2.6) and Pacific Island people (RR = 4.0, 95% CI = 3.1-5.3) were significantly more likely than people of European descent to acquire SAB, but not to die from the infection. Fifty per cent of cases were community acquired. A source was identified for 85%: intravenous catheter (31%), primarily hospital acquired, and skin/soft tissue (22%), primarily community acquired, were the most common foci. The 30-day mortality was 19%, 83% of whom died within 2 weeks. Risk factors for a poor outcome were: increasing age above 60, female sex (RR = 1.4, 95% CI = 1.0-2.1), diabetes mellitus (RR = 1.5, 95% CI = 1.0-2.4), immunosuppression (RR = 1.5, 95% CI = 1.0-2.4), pre-existing renal impairment (RR = 1.8, 95% CI = 1.2-2.7), malignancy (RR= 2.2, 95% CI = 1.4-3.5), lung as a source (RR = 2.8, 95% CI = 1.9-4.2) and unknown source (RR = 2.3, 95% CI = 1.5 3.3). Mortality was also accurately predicted by two multifactor scoring systems. There was a low rate of methicillin resistance (5%). CONCLUSIONS: Staphylococcus aureus bacteraemia is more likely to occur in certain ethnic groups, while mortality is associated with other identifiable risk factors and continues to be high. Intravenous catheters remain the most common and most preventable cause of SAB. PMID- 11480486 TI - When the rehabilitation ideal fails: a study of parental rights termination. AB - This study explored the circumstances of parents whose extreme neglect and abuse of their young children justified the drastic state action of termination of their parental rights. Records of 97 children age 6 and under whose parental rights were terminated between 1991 and 1997 in Minnesota were reviewed. Profiles of parents and children were drawn from these data, and a "risk pool" of parents whose children became wards of the state was identified. Guidelines are drawn from this "risk pool" that may help alert practitioners to those parents who are unlikely to safely maintain their children. Questions and implications for policy and practice are highlighted. PMID- 11480487 TI - Substance-abusing child welfare parents: treatment and child placement outcomes. AB - The authors present findings from their study of 167 child welfare parents referred for substance abuse assessments. Relationships between gender, prior treatment, court-ordered intervention, significant others' support, and treatment and placement outcomes are examined. Findings indicate significant others' support positively influences all outcomes while court-ordered intervention is not predictive. Prior treatment is associated with continued substance abuse. Gender differences exist for assessment completion and several client characteristics. Implications for practice are drawn. PMID- 11480488 TI - Paternal involvement in kinship foster care services in one father and multiple father families. AB - Using data from case records and from questionnaires completed by caseworkers, this article describes contact between 132 fathers of children in kinship foster care and their caseworkers over a period of 12 months, and the fathers' involvement in permanency planning for their children. The data indicate that most fathers had no contact with the caseworkers during the period under study and had never participated in planning. Analysis revealed that paternal involvement varied significantly by the child's family composition. Fathers of two or more children from a one-father family were most involved, while fathers of one child from a multiple-father family were least involved. Possible explanations for the findings are identified, and implications for practice and research are presented. PMID- 11480489 TI - Thiomersal in vaccines: is removal warranted? AB - The mercury-based vaccine preservative thiomersal has come under scrutiny in recent months because of its presence in certain vaccines that provide the foundation of childhood immunisation schedules. Over the past decade new vaccines have been added to the recommended childhood schedule, and the relatively smaller bodyweight of infants has led to concern that the cumulative exposure of mercury from infant vaccines may exceed certain guidelines for the human consumption of mercury. In the US, government agencies and professional societies have recently recommended that thiomersal be removed altogether from vaccines. Some involved in developing vaccine policy feel that the evidence to support these safety concerns has not risen to the level required for such a response. This apparent divergence of opinion has left healthcare professionals and the public with uncertainty about the potential health effects from low level exposure to thiomersal as well as the necessity of removing thiomersal from vaccines. At present, scientific investigation has not found conclusive evidence of harm from thiomersal in vaccines. As a precautionary measure, efforts are under way to remove or replace thiomersal from vaccines and providers should anticipate the availability of more vaccine products that are thiomersal-free over the coming years. PMID- 11480490 TI - Is gender a risk factor for adverse drug reactions? The example of drug-induced long QT syndrome. AB - Drug-induced torsade de pointes is a rare life-threatening adverse drug reaction (ADR) which is strongly influenced by gender. Drugs that prolong cardiac repolarisation include antiarrhythmics, gastrokinetics, antipsychotics, antihistamines and antibacterials. Such drugs share the potential to block cardiac voltage-gated potassium channels, particularly the rapid component (I(Kr)) of the delayed rectifier potassium current (I(K)). By doing so, such drugs usually, but not always, prolong the QT interval. Even if the electrocardiographic signs are subdued, the underlying blockade of I(Kr) current may precipitate the occurrence of arrhythmia. Women are perceived to be more prone to ADRs than men. Such a propensity may result from gender-associated differences in drug exposure, in the number of drugs prescribed (polypharmacy), in drug pharmacology, as well as from possible differences in the way the adverse event is perceived. A prolonged QT interval on the electrocardiogram (time that elapses from the onset of the cardiac ventricular depolarisation to the completion of its repolarisation) is associated with the occurrence of torsade de pointes and related ventricular arrhythmias. The QT interval is influenced by heart rate, autonomic nervous system, electrolyte disturbances and above all, drugs that block potassium channels. Two-thirds of the cases of drug-induced torsade de pointes occur in women. Therefore, this adverse effect represents a perfect example of gender differences impairing women's health. Clinical and experimental studies show that female gender is associated with a longer corrected QT interval at baseline and a greater response to drugs that block I(Kr), both of which facilitate the emergence of arrhythmia. This results most likely from a specific regulation of ionic channel expression (potassium, calcium, etc) by sex steroids, even though nongenomic effects may play a role as well. Estrogens facilitate bradycardia-induced prolongation of the QT interval and the emergence of arrhythmia whereas androgens shorten the QT interval and blunt the QT response to drugs. Hence, underlying genetic defects of potassium channels that may be asymptomatic in normal conditions, may precipitate drug induced arrhythmia in women more frequently than in men. Even in the presence of a drug that mildly blocks I(Kr) and seldom prolongs the QT interval, women are still more prone to drug-induced torsade de pointes, due to their reduced cardiac 'repolarisation reserve'. This is an important aspect of I(Kr) blockade to be aware of during the development of new drugs. PMID- 11480491 TI - Tolerability of postexposure antiretroviral prophylaxis for occupational exposures to HIV. AB - A substantial body of evidence provides support (but not definitive proof of efficacy) for the use of antiretroviral agents as postexposure prophylaxis for occupational exposures to HIV in the healthcare workplace. Despite the lack of definitive evidence of the efficacy of these agents in this setting, over the past decade this intervention has become the standard of care for healthcare workers who sustain occupational exposures to HIV. Administration of these agents -even for a relatively short 28-day postexposure course--is often fraught with difficulty. All of the agents currently used for postexposure prophylaxis regimens have substantial adverse effects, and significant adverse effects occur in more than two-thirds of individuals electing prophylaxis. This manuscript reiterates current US Federal Government guidelines for the administration of postexposure prophylaxis, specifically noting that zidovudine plus lamivudine (with or without a protease inhibitor) remains the recommended regimen. The paper summarises the significant toxicities associated with nucleoside reverse transcriptase inhibitors (primarily nausea, vomiting, diarrhoea and bone marrow suppression), non-nucleoside reverse transcriptase inhibitors (rash, fever, gastrointestinal symptoms and hepatitis, including hepatic decompensation necessitating liver transplantation) and protease inhibitors (nausea, vomiting, diarrhoea, abdominal pain, hyperglycaemia, hyperlipidaemia, headache and anorexia). As a class, the antiretroviral agents have an extraordinary number of drug interactions. The non-nucleoside reverse transcriptase inhibitors and the protease inhibitors are metabolised through the cytochrome P450 pathway, and the effects of concomitant administration of protease inhibitors with other agents in the same class are discussed, as well as the effects of concomitant administration of protease inhibitors with non-nucleoside agents. The potential for numerous and medically risky drug interactions emphasises the importance of planning antiretroviral prophylaxis in consultation with practitioners or clinical pharmacists who are skilled in the use of these agents and knowledgeable about the potential for significant drug interactions that could either reduce the benefit of prophylaxis or increase the potential for toxicity. Another common problem encountered by individuals managing postexposure prophylaxis programmes relates to the administration of chemoprophylaxis to a pregnant healthcare worker who has sustained an occupational exposure to HIV. We address what is known about the potential for toxicity and emphasise the recently published warning concerning the deaths of pregnant women and their offspring from lactic acidosis while receiving regimens containing stavudine and didanosine. PMID- 11480492 TI - Drug-induced angioedema without urticaria. AB - Angioedema without urticaria is a clinical syndrome characterised by self limiting local swellings involving the deeper cutaneous and mucosa tissue layers. Most occurrences of angioedema respond to treatment with a histamine H1 receptor blocker (antihistamine) because they are an allergic or parallergic reaction. A small number of cases do not respond to antihistamine treatment. Such cases tend to occur in patients with deficiency or dysfunction of the inhibitor of the first component of the complement (C1-INH), but more rarely can occur in patients with other conditions and as an adverse drug reaction. Angioedema is well documented in patients taking ACE inhibitors. Considering that 35 to 40 million patients are treated worldwide with ACE inhibitors, this drug class could account for several hundred deaths per year from laryngeal oedema. ACE inhibitors certainly do not mediate angioedema through an allergic or idiosyncratic reaction. For this reason the relationship with this drug is often missed and consequently quite underestimated. Rare instances of angioedema have also been reported with angiotensin II receptor antagonists. This adverse effect seems to occur less frequently with angiotensin II receptor antagonists than with ACE inhibitors. However, we do not know whether this adverse effect has the same mechanism with the 2 classes of medications. Some cases of severe angioedema have been recently reported after treatment with fibrinolytic agents. Scattered reports suggest the possibility of angioedema associated with the use of estrogens, antihypertensive drugs other than ACE inhibitors, and psychotropic drugs. Angioedema can also occur with nonsteroidal anti-inflammatory drugs. Prevention of angioedema relies first on the patient history. Estrogen and ACE inhibitors should be avoided in a patient with congenital or acquired C1-INH deficiency. In the case of ACE inhibitors, the appearance of angioedema following long term treatment does not lessen the probability that such an agent could be the cause. The most important action to take in a patient with suspected drug-induced angioedema is to discontinue the pharmacological agent. Epinephrine (adrenaline), diphenydramine and intravenous methylprednisolone have been proposed for the medical management of airway obstruction, but so far no controlled studies have demonstrated their efficacy. If the acute airway obstruction leads to life-threatening respiratory compromise an emergency cricothyroidotomy must be performed. PMID- 11480494 TI - Safety and efficacy of long term esomeprazole therapy in patients with healed erosive oesophagitis. AB - OBJECTIVE: To evaluate the safety and tolerability of long term treatment with esomeprazole in patients with healed erosive oesophagitis, and to describe its efficacy in the maintenance of healing. DESIGN AND SETTING: US multicentre, noncomparative, nonblind study. PATIENTS AND PARTICIPANTS: 807 patients with endoscopically confirmed healed erosive oesophagitis. METHODS: Patients received esomeprazole 40 mg once daily for up to 12 months. Adverse events and clinical laboratory tests were assessed over the study period. Endoscopy was performed at the final visit of the antecedent healing trials and at months 6 and 12 of the current safety trial; gastric biopsies were obtained at the initial visit of the healing trials and at the end of the safety trial. RESULTS: 80.9% of patients completed 6 months of treatment; 76.6% completed 12 months of treatment. There were no serious drug-related adverse events. Diarrhoea, abdominal pain, flatulence, and headache were the only treatment-related adverse events reported by >3% of patients. Mean changes in laboratory measures were generally small and not clinically meaningful. Plasma gastrin levels increased, as expected, and reached a plateau after 3 months. No changes in gastric histological scores were noted in the majority of patients. Evaluation of gastric biopsies revealed an overall decline in chronic inflammation and atrophy. Intestinal metaplasia findings remained essentially unchanged. Life table estimates of maintenance of healing were 93.7% [95% confidence interval (CI) 92.0 to 95.5%] at 6 months and 89.4% (95% CI 87.0 to 91.7%) at 12 months. CONCLUSIONS: Daily treatment with esomeprazole 40 mg for up to 1 year in patients with healed erosive oesophagitis was generally well tolerated and effective. No safety concerns arose. PMID- 11480493 TI - Corticosteroid-Induced osteoporosis: detection and management. AB - Corticosteroid-induced osteoporosis is a major cause of morbidity and is the leading secondary cause of osteoporosis today. Unfortunately, despite this knowledge, patients receiving corticosteroid therapy are often not offered any preventative treatment. Recent research has focused attention on the critical role the osteoblast has played in the pathophysiology of corticosteroid-induced osteoporosis. In addition to an initial increase in bone resorption, there is evidence that corticosteroids induce osteoblast and osteocyte apoptosis and as a result are important contributors to bone loss. Interesting work has suggested that the bisphosphonates and calcitonin may help to prevent osteoblast apoptosis from occurring. Large scale randomised controlled trials have also been completed with a variety of therapeutic agents. Of the many different therapies, it is now clear that the bisphosphonates have the greatest evidence to support their use. Increases in bone mineral density when compared with a control group, not only at the spine but also at the hip, have been demonstrated. These studies have shown clinically significant reductions in vertebral fracture rates seen for the most part in postmenopausal women. Other therapies may well be effective, as evidenced by maintenance of bone mass in the spine; however, maintenance of bone mass in the hip and reductions in fracture rate have yet to be demonstrated for many of these therapies. Given our current knowledge and the evidence that is outlined in this review, it is hoped that patients who require therapy with corticosteroids for more than 3 months will be offered appropriate preventative treatment. PMID- 11480495 TI - A multiple center study of multiple chemical sensitivity syndrome. AB - The lack of widely accepted, standardized, clinical and epidemiologic criteria for Multiple Chemical Sensitivity syndrome has led to confusion about the identification of the condition and has slowed pertinent research. In this article, the authors evaluated the psychometric properties of 2 sets of clinical/epidemiologic criteria for Multiple Chemical Sensitivity syndrome. In this cross-sectional survey of 1,166 patients who visited outpatient occupational, otolaryngology, allergy, and clinical ecological clinics, the authors used the aforementioned sets of criteria to (a) estimate the prevalence of the syndrome in these varied samples and (b) compare the current diagnostic practices of traditional physician specialists with those of clinical ecologists. The authors used a patient-completed questionnaire to assess the medical, psychosocial, and psychological status of patients who reported multiple chemical sensitivities. This approach enabled the formulation of 6 domains, which represented commonly observed characteristics of the syndrome. The authors used a physician-completed questionnaire to collect diagnoses of Multiple Chemical Sensitivity syndrome and other medical conditions. Domains, which were operationalized by the questionnaire and comprised the 2 sets of criteria for identification of the Multiple Chemical Sensitivity syndrome, had test-retest reliabilities that exceeded .75 and estimates of internal consistency that ranged between .59 and .94. Evidence of construct and face validity was considered acceptable. The overall clinic-based prevalences of Multiple Chemical Sensitivity syndrome, based on 6 and 4 domains, were 7% and 23%, respectively. Regardless of the identifying set of criteria used, physicians' diagnoses had relatively low sensitivities (range = 6-50%) and relatively high specificities (range = 82-99%). The study data suggested that the domains operationalized by the questionnaire had reasonable psychometric characteristics. Study data also support the fact that Multiple Chemical Sensitivity syndrome is often overlooked--even by those physicians who treat it most frequently--and that use of both sets of objective criteria for identifying the syndrome would greatly improve the sensitivity of physician diagnoses. PMID- 11480496 TI - Residential radon and risk of lung cancer in an Italian alpine area. AB - To evaluate whether residential radon exposure explains the excess mortality for lung cancer in an Italian alpine valley with high natural radioactivity, the authors conducted a population-based case-control study on 138 deceased cases and 291 sex- and year-of-birth-matched controls. Year-long alpha-track measurements of radon were performed in the most recent residence, and information about occupational history and lifetime smoking habits was obtained. The authors adjusted for smoking, and radon was associated with lung cancer risk among men: compared with a radon level of < 40 becquerels (Bq) per cubic meter (m3), the odds ratios for 40-76 Bq/m3, 77-139 Bq/m3, 140-199 Bq/m3, and 200+ Bq/m3 were 2.1, 2.0, 2.7, and 1.4, respectively. The association between radon and lung cancer, as determined with a multiplicative model, was found only among male smokers. PMID- 11480497 TI - Effects of aerosol-vapor JP-8 jet fuel on the functional observational battery, and learning and memory in the rat. AB - To determine whether JP-8 jet fuel affects parameters of the Functional Observational Battery (FOB), visual discrimination, or spatial learning and memory, the authors exposed groups of male Fischer Brown Norway hybrid rats for 28 d to aerosol/vapor-delivered JP-8, or to JP-8 followed by 15 min of aerosolized substance P analogue, or to sham-confined fresh room air. Behavioral testing was accomplished with the U.S. Environmental Protection Agency's Functional Observational Battery. The authors used the Morris swim task to test visual and spatial learning and memory testing. The spatial test included examination of memory for the original target location following 15 d of JP-8 exposure, as well as a 3-d new target location learning paradigm implemented the day that followed the final day of exposure. Only JP-8 exposed animals had significant weight loss by the 2nd week of exposure compared with JP-8 with substance P and control rats; this finding compares with those of prior studies of JP-8 jet fuel. Rats exposed to JP-8 with or without substance P exhibited significantly greater rearing and less grooming behavior over time than did controls during Functional Observational Battery open-field testing. Exposed rats also swam significantly faster than controls during the new target location training and testing, thus supporting the increased activity noted during Functional Observational Battery testing. There were no significant differences between the exposed and control groups' performances during acquisition, retention, or learning of the new platform location in either the visual discrimination or spatial version of the Morris swim task. The data suggest that although visual discrimination and spatial learning and memory were not disrupted by JP-8 exposure, arousal indices and activity measures were distinctly different in these animals. PMID- 11480498 TI - The role of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and regulated on activation, normal T-cell expressed and secreted (RANTES) in the relationship between air pollution and asthma among children. AB - To evaluate the role of adhesion molecules and chemokines in the relationship between air pollution and asthma, the authors determined the following in 230 children who lived in 4 communities in Japan that had different levels of air pollution: serum concentrations of soluble intercellular adhesion molecule-1 (sICAM-1); soluble vascular cell adhesion molecule-1 (sVCAM-1); regulated on activation, normal T-cell expressed and secreted (RANTES); and total immunoglobulin E (IgE). Children with asthma or who exhibited wheezing (n = 115) and nonasthmatic children (n = 115) were studied. Serum concentrations of sICAM 1, sVCAM-1, and RANTES were increased significantly in asthmatic children, compared with nonasthmatic children. Nonasthmatic children with serum IgE levels that were greater than or equal to 250 IU/ml had significantly lower concentrations of sICAM-1 and sVCAM-1 than the other children. The geometric means of sICAM-1 increased as air pollution increased. The results of this study suggest that adhesion molecules or chemokines are associated with asthma and that ICAM-1 may play an important role in the relationship between air pollution and the occurrence of asthma. PMID- 11480499 TI - Acute asthma exacerbations and air pollutants in children living in Belfast, Northern Ireland. AB - The incidence of childhood asthma, a common condition, is on the rise worldwide. Despite reductions in the emission of urban smoke, traffic pollution is now a major worldwide problem. Belfast, Northern Ireland, is an old industrial city with major pollution problems. In this study, the authors investigated the rates of acute asthma admissions to Belfast's major children's emergency department. The admissions were studied, relative to day-to-day fluctuations in thoracic particulate matter, sulfur dioxide, nitrogen dioxide, nitric oxide, oxides of nitrogen, ozone, carbon monoxide, benzene, temperature, and rainfall. Daily admissions for acute asthma at the emergency department of the Royal Belfast Hospital and average daily pollution were recorded for the 3-yr period between January 1, 1993, and December 31, 1995. The authors used Poisson regression to assess independent association(s). Individually, small associations were seen for thoracic particulate matter (relative risk = 1.10), sulfur dioxide (relative risk = 1.09), nitrogen dioxide (relative risk = 1.11), nitric oxide (relative risk = 1.07), oxides of nitrogen (relative risk = 1.10), carbon monoxide (relative risk = 1.07), and benzene (1.14); no associations were noted between meteorological factors (temperature and rainfall) or ozone and asthma emergency-department admissions. The authors adjusted for the aforementioned parameters, and benzene level was the only variable associated independently with asthma emergency department admissions in children. Benzene may be a more reliable method of measuring exposure to vehicle exhaust emissions than measurements of other pollutants. PMID- 11480500 TI - Dietary antioxidants and ozone-induced bronchial hyperresponsiveness in adults with asthma. AB - Ozone exposure aggravates asthma, as has been demonstrated in both controlled exposures and epidemiologic studies. In the current double-blind crossover study, the authors evaluated the effects of dietary antioxidants (i.e., 400 IU vitamin E/500 mg vitamin C) on ozone-induced bronchial hyperresponsiveness in adult subjects with asthma. Seventeen subjects were exposed to 0.12 ppm of ozone or to air for 45 min during intermittent moderate exercise. Bronchial hyperresponsiveness was assessed with 10-min sulfur dioxide (i.e., 0.10 ppm and 0.25 ppm) inhalation challenges. Subjects who were given dietary antioxidants responded less severely to sulfur dioxide challenge than subjects given a placebo (i.e., forced expiratory volume in the 1st sec: -1.2% vs. 4.4%, respectively; peak flow: +2.2% vs. -3.0%, respectively; and mid-forced expiratory flow: +2.0% vs. -4.3%, respectively). Effects were more pronounced when subjects were grouped by response to sulfur dioxide at the screening visit. The results suggest that dietary supplementation with vitamins E and C benefits asthmatic adults who are exposed to air pollutants. PMID- 11480501 TI - Infecundity and consumption of polychlorinated biphenyl-contaminated fish. AB - Biologic capacity for reproduction, or fecundity, may be threatened by environmental contaminants, especially compounds capable of disrupting endocrine pathways. Telephone interviews that focused on reproductive events were conducted with female members of the New York State Angler Cohort Study who became pregnant between 1991 and 1993 and who reported known time to pregnancy (N = 895; 73%). Consumption of polychlorinated biphenyl-contaminated Lake Ontario sportfish and other factors were ascertained in 1991. The authors classified the women as follows: (a) fecund (time to pregnancy < or =12 cycles; n = 723); (b) having resolved infecundity (time to pregnancy > 12 cycles; n = 81); or (c) having unresolved infecundity (time to pregnancy > 12 cycles without pregnancy; n = 94). Adjusted odds ratios for duration of fish consumption for both resolved and unresolved infecundity were elevated (1.46 and 1.19, respectively), although confidence intervals included unity. Frequency of recent fish consumption was associated with an increased risk for select categories, although confidence intervals included one. PMID- 11480502 TI - Current perception thresholds in vibration-induced neuropathy. AB - The authors evaluated the usefulness of current perception threshold testing for the assessment of vibration-induced neuropathy. The study population comprised 20 male controls and 59 males with hand-arm vibration syndrome. Current perception threshold at three test frequencies (i.e., 5 Hz, 250 Hz, and 2,000 Hz) was determined on the distal phalanges of the index and little fingers. Large myelinated fibers, small myelinated fibers, and unmyelinated fibers were evaluated selectively at 2,000-Hz, 250-Hz, and 5-Hz frequencies, respectively. The vibration-induced neuropathy groups had significantly increased current perception thresholds in both the index and little fingers. There was a significant increase in current perception threshold for the myelinated fibers, but not the unmyelinated fibers. The diagnostic sensitivity for vibration-induced neuropathy was relatively high. It appeared that current perception threshold evaluation was useful for the assessment of vibration-induced neuropathy. PMID- 11480503 TI - Systemic nicotine exposure in tobacco harvesters. AB - Several epidemics of nicotine intoxication have been described among tobacco harvesters; however, little is known about nicotine absorption under typical working conditions. To assess systemic nicotine absorption during a regular working shift, the authors performed an observational field study. Included in the study were 10 healthy, nonsmoking, female tobacco harvesters and a control group of 5 healthy, nonsmoking, female hospital workers. Nicotine and cotinine were measured in sequential samples of blood and urine during a regular workshift. Blood nicotine levels rose from a nadir value of 0.79 +/- 0.12 ng/ml to a peak value of 3.45 +/- 0.84 ng/ml (p < .05 [Tukey's modified t test]) in the exposed group. In the control group, levels were stable at 0.1 +/- 0.1 ng/ml (p < .01). Moreover, the mean blood nicotine level measured 3 mo following the end of exposure in 6 of 10 exposed subjects was 0.24 +/- 0.12 ng/ml (p < .01). Corresponding higher values of urine nicotine and urine cotinine were observed in the exposed versus control group (comparative p values were < .01 and < .05, respectively). Overall, tobacco harvesters absorbed approximately 0.8 mg of nicotine daily. Given that nicotine can induce adverse health effects, the authors believe that prevention of nicotine absorption in tobacco harvesters should be sought and that workers should be informed about occupational risks. PMID- 11480504 TI - Short-term adverse health effects in a community exposed to a large polyvinylchloride plastics fire. AB - The purpose of this study was the documentation of the short-term morbidity and mortality experiences of an urban community exposed to the airborne byproducts of a large polyvinylchloride plastics fire. The authors administered a survey to representatives of each household who had lived in an area evacuated during the fire. A time-series analysis was performed on emergency room visits and admissions for all hospitals in the city. Chloracne surveillance was instituted. Sixty-two percent of the individuals surveyed from the evacuation area reported no health concerns or symptoms related to the fire. Thirty-eight percent of the residents reported symptoms, and less than 2% of those surveyed reported that they sought medical attention for their health concerns. There was no evidence of increased hospital admissions or emergency room use during and immediately following the fire. No cases of chloracne were reported, and no deaths or serious injuries occurred during the fire. Polyvinylchloride plastics recycling plants pose potential health hazards to civilian populations. Public health authorities should be prepared to assess population health status rapidly and to disseminate relevant health information in a timely way during a crisis. PMID- 11480505 TI - Electrolyte loss in sweat and iodine deficiency in a hot environment. AB - The authors studied electrolyte loss from profuse sweating in soccer-team players and evaluated the relationship between this source of iodine loss and iodine deficiency. Thirteen male soccer-team players and 100 sedentary students from the same high school were evaluated for 8 d, during which the players were training. The authors analyzed 208 sweat samples to determine losses of iodine, sodium, potassium, and calcium in sweat. Excretion of urinary electrolytes by the subjects was also measured. The mean losses of iodine, sodium, potassium, and calcium in sweat following a 1-hr game were 52 microg, 1,896 mg, 248 mg, and 20 mg, respectively; the ratios of sweat loss to urinary daily loss of the four electrolytes were 0.75, 0.2, 1.88, and 0.92, respectively. Urinary iodine was significantly (p < .02) lower than the normal level of 50 microg/gm creatinine in 38.5% of the soccer players, compared with 2% of the sedentary students. Forty six percent of the players had Grade I goiter, compared with a mere 1% of the sedentary students (p < .01). The results of the study suggest that loss of iodine through profuse sweating may lead to iodine deficiency, and loss of electrolytes through sweating may have a dietary significance for heat-stressed individuals or for individuals who perform heavy workloads. PMID- 11480506 TI - Lead and cadmium exposure from contaminated soil among residents of a farm area near an industrial site. AB - In this study, the authors determined the degree of lead and cadmium exposure in a population that resided in an area with contaminated soil. The extent of exposure from soil pollution was also assessed. Lead and cadmium concentrations in blood of children and adults who resided in the contaminated area were measured, and cadmium concentration in urine of adults was also determined. An adult control group was recruited from a nonpolluted area. The mean blood lead level in adults who resided in the polluted area was 9.8 microg/dl, compared with a mean level of 6.8 microg/dl in controls (p = .004). Urinary cadmium levels were well below the level associated with onset of symptoms, but the differences between levels in residents of the contaminated area (0.54 microg/gm creatinine) and levels in the controls (0.37 microg/gm creatinine) indicated that life-long cadmium exposure had been higher among the residents of the contaminated area (p = .086). The mean blood lead level and mean blood cadmium level in children were 5.2 microg/dl (maximum = 7.90 microg/dl) and 0.10 microg/l, respectively. Lead in soil accounted in large part for the differences in blood lead levels in children; however, blood cadmium levels were not associated with soil cadmium levels, but, rather, with consumption of home-grown vegetables. PMID- 11480507 TI - Urinary platinum levels associated with dental gold alloys. AB - Platinum concentrations were determined in 50 urine and 20 saliva samples obtained from 50 subjects who had gold dental restorations. In addition, 42 urine and 35 saliva samples were collected from subjects who did not have gold dental restorations. Subjects with gold alloys had significantly (p < .001) higher urinary platinum excretion (mean = 11.9 +/- 8.5 ng/gm creatinine, range = 1.9 45.8 ng/gm creatinine) than controls (mean = 6.2 +/- 3.2 ng/gm, range = 1.9-14.4 ng/gm creatinine). Mean saliva concentrations were significantly higher in subjects with dental gold alloys (526 pg/gm vs. 8.5 pg/gm; p < .001). A laboratory test with 5 commercially available dental gold/platinum alloys showed that 0.1% sodium chloride mobilized platinum within 1 hr (i.e., 1-18 pg/ml) of its introduction. In conclusion, dental gold/platinum alloys appear to be the main source for urinary platinum excretion from the occupationally unexposed population. PMID- 11480508 TI - Prospective evaluation of the effects of prostatectomy on symptoms and quality of life. AB - AIMS: To evaluate prospectively urinary symptoms and quality of life before and after prostate surgery, among men who had been on a waiting list for elective transurethral prostate resection (TURP). METHODS: Face-to-face interviews with 47 men before and after prostate surgery using the urinary symptom specific American Urological Association Urinary Symptom Index (AUA-7), the Short Form 36 Health Survey (SF-36), and questions relating to the effects of urinary symptoms on men's lives and acceptable waiting times for surgery. RESULTS: The mean AUA-7 score before surgery for all participants was 22.2 (range 5-34) and after surgery improved to a mean of 7.3 (range 1-28). Men with severe urinary symptoms before surgery experienced the greatest improvement in symptoms after surgery, compared with the mild/moderate symptom group. Significant improvements occurred in three of the eight SF-36 dimensions following surgery. Acceptable waiting times for surgery differed according to the severity of mens' urinary symptoms. CONCLUSIONS: Efforts to develop methods for prioritisation of access to surgery are supported. Men with mild or moderate symptoms may reasonably delay surgery, with the expectation that, if they develop more severe symptoms, surgery will be helpful. Because of the risks as well as the benefits of prostate surgery, the use of illustrative vignettes may be useful for clinicians advising patients. PMID- 11480509 TI - Surveillance of sexually transmitted infections in New Zealand, 1998. AB - AIMS: To describe the surveillance and epidemiology of sexually transmitted infections (STIs) in New Zealand. METHODS: Sexual health clinics submitted STI data to the Institute of Environmental Science and Research (ESR). Infection rates were calculated by dividing the number of diagnoses by the number of total clinic visits. Because the denominator used to calculate infection rates changed in 1998, STI rates in 1998 cannot be directly compared with previous years and case numbers were used to identify recent trends. RESULTS: In 1998, genital warts was the most commonly diagnosed STI (4.7%), followed by chlamydia (3.0%) and genital herpes (1.0%). Approximately two-thirds of gonorrhoea, chlamydia and genital warts diagnoses were in people aged less than 25 years. Chlamydia rates were 7.3% in Maori, 7.1% in Pacific Island people, and 2.1% in European. Gonorrhoea rates were 1.6% in Maori, 1.9% in Pacific Island people and 0.2% in European. The number of chlamydia and gonorrhoea cases increased between 1995 and 1998. CONCLUSIONS: The reporting of data by age, sex and ethnicity has allowed a more useful evaluation of the incidence of STIs. The majority of STIs were diagnosed among young New Zealanders, and disproportionately high chlamydia and gonorrhoea infection rates were found among Maori and Pacific Island people. PMID- 11480510 TI - Pharmaceutical management in ProCare Health Limited. AB - AIMS: To review pharmaceutical budget holding and management in ProCare Health Limited by; describing budget holding strategies implemented in 1995/6, identifying prescribing savings achieved, analysing variation in prescribing behaviour and comparing the findings with experience elsewhere. METHODS: With 340 members, ProCare is one of the largest and most progressive of New Zealand's independent practitioner associations (IPAs). Data were obtained for the three years 1994 to 1996 to determine pharmaceutical expenditure against budget and against national trends, by member and general medical services (GMS) consultations. RESULTS: ProCare has established a classical, quality focussed pharmaceutical management strategy. Savings against the agreed budget was 9.5% comparing 1996 with 1995 but 5.7% compared, with national trends. Wide variation in per capita and per consultation costs was not reduced and was entirely explained by prescribing volumes not drug prices. CONCLUSIONS: The most important finding is that general practitioners (GPs), working collaboratively, can establish a strategy of clinical and corporate governance which may be exerting a wide ranging influence over clinical behaviour. Although there may be doubts about the actual levels of saving these appeared to be well in excess of the financial investment in the strategy. Greater savings appear possible with a focus on addressing the large and apparently inappropriate per capita prescribing volume variation between practices. Understanding and successfully addressing this variation will be one of the key issues facing the implementation of the government's primary health care strategy. PMID- 11480511 TI - Auditory and vestibular sequelae to traumatic brain injury: a pilot study. AB - AIM: To investigate the incidence of persisting auditory and vestibular sequelae in a group of 30 young adults recovering from Traumatic Brain Injury (TBI). METHODS: 30 participants, aged 21-45 years, with TBI suffered 19 months to 27 years previously, underwent a semi-structured interview and pure-tone hearing test in their home. Participants who failed the hearing screen then undertook a more comprehensive audiological evaluation. RESULTS: A variety of sequelae to TBI were reported. These were interpreted as tinnitus (53%), vestibular dysfunction (83%), abnormal facial sensory symptoms (27%) and intolerance to loud/sudden noises (87%). Ten (33%) participants demonstrated significant sensorineural hearing impairment in addition to speech recognition performance significantly worse than would have been predicted from their hearing impairment. CONCLUSIONS: Findings from this study will be of benefit to health professionals working in this area of rehabilitation as they seek to provide functional assessments and devise programmes to treat the often devastating auditory processing problems of people recovering from TBI. PMID- 11480512 TI - The views of Otago urban and rural general practitioners on mental health services. AB - AIM: To examine the views of Otago general practitioners (GPs) about local mental health services and their role in providing such health care. METHODS: A questionnaire was distributed to all GPs working in Otago. RESULTS: 100 replies (59%) were received. There were large differences between GPs in the estimated numbers of patients with psychiatric conditions they were seeing. 85% were keen on sharing management of patients with mental health services. Average confidence levels for diagnosis and management of depression and anxiety were good. GPs felt less confident about psychosis, somatisation, eating and personality disorders. Obstacles to GPs doing more mental health work included time, cost, access to specialist services and training. On balance, GPs viewed the present mental health services as adequate. CONCLUSIONS: GPs appeared prepared to do more mental health work if the barriers of cost and time could be overcome and they were provided with adequate education and backup from specialised services. PMID- 11480513 TI - A prospective study of immunisation for a cohort of children. AB - AIMS: To determine rates of immunisation by 24 months of age, the number of times children were recalled and the cost of immunisation for a cohort of children in general practices in Wellington. METHODS: A prospective study of 979 children registered with 27 general practices. Children in the cohort were followed from 9 24 months old. Data collected included immunisation status, the number of times children were recalled and demographic data. RESULTS: At the end of the study period (when children reached 24 months of age) 803 (82%) remained with the practices and 176 (18%) had left. At 24 months 724 (74%) of the total cohort and 685 (85.3%) of registered children who stayed with the practice were fully immunised for the early childhood vaccinations. 54% of the cohort were fully immunised after a standard recall process. The average cost per child immunised was $13.33. CONCLUSION: It is possible to achieve high rates of full immunisation in children registered with a general practice using an effective system of facilitation and support. PMID- 11480514 TI - The future of specialist psychiatric services in rural New Zealand. PMID- 11480515 TI - Marching from the margin: a health vision for Pacificans of Aotearoa. PMID- 11480516 TI - Breast screening--time for review. PMID- 11480517 TI - The relationship between the occurrence of undifferentiated bovine respiratory disease and titer changes to bovine coronavirus and bovine viral diarrhea virus in 3 Ontario feedlots. AB - Serological evidence of previous viral exposure (titer at arrival) and current viral exposure (titer increase) during a 28-day study period, was used to determine if bovine coronavirus (BCV) or bovine viral diarrhea virus (BVDV) was associated with the occurrence of undifferentiated bovine respiratory disease (UBRD) in feedlot calves. Neutralizing antibody titers to BCV and BVDV were determined for 852 animals from 3 Ontario feedlots. Calves at 2 of the 3 feedlots (n = 753) received a modified live 4-way viral vaccine containing BVDV. On arrival at the feedlots, 90% of animals were seropositive for BCV, while 39% of animals were seropositive for BVDV. This evidence of previous exposure to both viruses was associated with reduced subsequent UBRD risk. Evidence of exposure to BCV during the study period was common, as 50% of animals showed a 16-fold or greater titer increase; however, treatment for UBRD was not associated with titer change. Although the majority of animals were vaccinated for BVDV at arrival, within a feedlot, animals treated for UBRD had larger titer increases to BVDV than non-treated animals. Based on our findings we infer that BCV was not causally related to UBRD occurrence, however consistent with other literature, BVDV may be causally related to UBRD occurrence. PMID- 11480518 TI - The relationship between the occurrence of undifferentiated bovine respiratory disease and titer changes to Haemophilus somnus and Mannheimia haemolytica at 3 Ontario feedlots. AB - The association between exposure to Haemophilus somnus and Mannheimia haemolytica (formerly Pasteurella haemolytica) and the risk of undifferentiated bovine respiratory disease (UBRD) was investigated using serological evidence of exposure coupled with a factorial design vaccine field trial. Measures of previous exposure (titer at arrival) and current exposure (titer increase in the study period) to these agents were used. The vaccine field trial involved systematic allocation of animals into groups that received either a M. haemolytica vaccine, an H. somnus vaccine, a combined M. haemolytica and H. somnus vaccine, and an unvaccinated control group. Serum was collected from the 852 animals enrolled to determine titers to H. somnus, M. haemolytica, bovine coronavirus and bovine viral diarrhea virus. Vaccination with H. somnus in combination with M. haemolytica and with M. haemolytica alone reduced the risk of UBRD. The odds ratio for vaccination with H. somnus alone and UBRD risk suggested some sparing effect, but the 95% confidence limits included unity. There was no association between serological evidence of concurrent exposure to M. haemolytica and UBRD occurrence. There was an association between titer change to H. somnus and UBRD risk. However, the association changed with time of BRD treatment; animals diagnosed and treated for UBRD on or after day 10 showed little evidence of exposure to H. somnus, despite evidence of natural H. somnus exposure in the unvaccinated group. The association between titer change to H. somnus and UBRD occurrence seen in this study may be a consequence of prolonged exposure to antibiotics, rather than a causal association. PMID- 11480519 TI - Prevalence, capsular type and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs in Korea. AB - This study was undertaken to determine the prevalence, capsular serotype, and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs. Capsular serotype and antimicrobial susceptibility were determined by coagglutination test and agar dilution minimum inhibitory concentration, respectively. Streptococcus suis was isolated from 55 of the 406 palatine tonsillar samples tested (13.8%) and 14 of the 29 sampled herds (48.3%). Of the 55 isolates recovered from slaughter pigs, 26 (47.3%) were untypeable. Of the remaining 29 isolates, capsular serotypes 9 (9 isolates) and 16 (4 isolates) were the most common, followed by capsular serotypes 4 (3 isolates) and 7 (3 isolates). Every capsulated isolate was typeable and no palatine tonsillar sample yielded more than one serotype. Most of isolates were susceptible to low concentrations (MIC90) of amoxicillin (2 microg/mL), ceftiofur (1 microg/mL), and penicillin (1 microg/mL). No correlation was found between antimicrobial susceptibility and capsular serotype. PMID- 11480520 TI - Decrease of the adhesion of Streptococcus suis serotype 2 mutants to embryonic bovine tracheal cells and porcine tracheal rings. AB - Streptococcus suis is an important swine pathogen that may be present in the tonsils of pigs that show no signs of illness. Because adhesion to host cells may be important in the carrier state, this study was undertaken to investigate adhesion to host cells by S. suis mutant strains defective in expression of a 39 kDa protein. Mutant strains of S. suis were generated by transposon Tn916 mutagenesis and were tested for adhesion to embryonic bovine tracheal cells and porcine tracheal rings. Compared with the parent strain, there was a significant reduction in adherence of 3 mutant strains to both bovine tracheal cells and porcine tracheal rings. PMID- 11480521 TI - Experimental colonization of piglets and gilts with systemic strains of Haemophilus parasuis and Streptococcus suis to prevent disease. AB - Haemophilus parasuis and Streptococcus suis are both major causes of losses during the nursery period, especially in herds using the segregated early weaning system. In this system, only a few piglets may be colonized with the herd's prevalent systemic strain, which results in infection of naive penmates late in the nursery. In view of these factors, the objectives of this study were: (1) to evaluate the early colonization of piglets with the farm's prevalent systemic strain of H. parasuis and S. suis as an alternative method for disease prevention; and (2) to evaluate 2 different protocols for experimental colonization: direct colonization of piglets and colonization of piglets through nose-to-nose contact with inoculated sows. Haemophilus parasuis and S. suis isolates recovered from diseased nursery pigs were characterized by the rep-PCR technique and the herd's prevalent strains were used for colonization. Piglets in the experimentally colonized groups were inoculated at 5 days of age by the oral route using a spray pump. Sows were colonized at 2 weeks prior to farrowing using a similar protocol. Although both colonization protocols were successful in getting the piglets colonized, direct inoculation of 5-day-old piglets with the herd's systemic strains of H. parasuis and S. suis tended to be more effective in reducing the morbidity and the mortality than the colonization of piglets by nose to-nose contact with inoculated sows. PMID- 11480522 TI - Host response to various treatments to reduce Salmonella infections in swine. AB - Host response was evaluated following the administration of various treatments, such as probiotics, prebiotics, and vaccination, to reduce Salmonella in swine. Response to the treatments were studied by the evaluation of phagocytosis rates by flow cytometry, by studying the activation of whole-blood phagocytes by bioluminescence, the production of IgA against S. Typhimurium, and by histopathology. Significant differences were observed in the activation of whole blood phagocytes in all groups of treated pigs (P = 0.0001). In SC54 vaccinated pigs, a significant reduction of Salmonella in the ileum was observed (P < 0.05) and the production of IgA against S. Typhimurium was higher in this group in comparison to uninfected control pigs (P = 0.0007). Furthermore, significant histopathological (P < 0.05) changes were observed in SC54 vaccinated pigs. Villus height and mucus and goblet cells density in the small intestine were reduced in vaccinated pigs in comparison to infected control pigs. Taken together, these findings suggest that SC54 vaccine can stimulate local immunity and reduce the presence of Salmonella in the ileum in swine. Use of SC54 vaccine should thus be considered in further field experiments. PMID- 11480523 TI - Penicillin concentrations in serum, milk, and urine following intramuscular and subcutaneous administration of increasing doses of procaine penicillin G in lactating dairy cows. AB - Eight healthy, non-pregnant, crossbred Holstein dairy cows (557-682 kg) within their first 3 months of lactation (13-21.5 kg of milk/day) were used. Cows were kept in tie stalls for the whole experiment. The 8 cows were randomly assigned to 2 (IM and SC) 4 x 4 balanced Latin square design experiments. Doses of procaine penicillin G (PPG) (300000 IU/mL) in each square were 7000, 14000, 21000 and 28000 IU/kg and were injected IM or SC once daily for 5 consecutive days. Volumes of PPG per site of injection never exceeded 20 mL. Blood was collected to determine the Cmax, Tmax, and AUC; urine and milk were also taken to measure the persistence of PPG in these fluids. Results show that serum Cmax and Tmax were only slightly affected by increasing the doses or the route of administration, whereas the AUC was linearly increased in relation to the dose injected in both modes of injection. In the urine, Cmax varied from 160 to 388 IU/mL and Tmax from 72-120 h during 5 consecutive days of PPG injection. A dose effect in Cmax was observed only for the IM route of administration and no variation (P > 0.05) was found between the IM and SC routes. Milk Cmax concentrations were only increased by the dose regimen in the IM group. At doses of 21000 and 28000 IU/kg, the IM group had a higher (P > 0.05) Cmax when compared with the SC groups. Milk PPG residues were not detectable over 96 h following the last IM injection, independently of the dose injected. However milk PPG residues were detected for up to 132 h following the last SC injection. These results show that when PPG is injected IM once daily in volumes not exceeding 20 mL/site at doses as high as 28000 IU/kg, the withdrawal period should be at least 96 h. Therefore, in the present model, there was no advantage to inject PPG by SC route to improve PPG kinetic parameters as the AUC, Cmax, or Tmax. PMID- 11480524 TI - Pharmacokinetics of orbifloxacin and its concentration in body fluids and in endometrial tissues of mares. AB - Pharmacokinetics and distribution of orbifloxacin into body fluids and endometrium was studied in 6 mares after intragastric (IG) administration at a single dose rate of 7.5 mg/kg body weight. Orbifloxacin concentrations were serially measured in serum, synovial fluid, peritoneal fluid, urine, cerebrospinal fluid, and endometrial tissues over 24 hours. Minimum inhibitory concentrations of orbifloxacin were determined for 120 equine pathogens over an 11-month period. The mean peak serum concentration (Cmax) was 2.41+/-0.30 microg/mL at 1.5 hours after administration and decreased to 0.17+/-0.01 microg/mL (Cmin) at 24 hours. The mean elimination half-life (t1/2) was 9.06+/ 1.33 hours and area under the serum concentration vs time curve (AUC) was 20.54+/ 1.70 mg h/L. Highest mean peritoneal fluid concentration was 2.15+/-0.49 microg/mL at 2 hours. Highest mean synovial fluid concentration was 1.17+/-0.28 microg/mL at 4 hours. Highest mean urine concentration was 536.67+/-244.79 microg/mL at 2 hours. Highest mean endometrial concentration was 0.72+/-0.23 microg/g at 1.5 hours. Mean CSF concentration was 0.46+/-0.55 microg/mL at 3 hours. The minimum inhibitory concentration of orbifloxacin required to inhibit 90% of isolates (MIC90) ranged from < or = 0.12 to > 8.0 microg/mL, with gram negative organisms being more sensitive than gram-positive organisms. Orbifloxacin was uniformly absorbed in the 6 mares and was well distributed into body fluids and endometrial tissue. At a dosage of 7.5 mg/kg once a day, many gram-negative pathogens, such as Actinobacillus equuli, Escherichia coli, Pasteurella spp., and Salmonella spp. would be expected to be susceptible to orbifloxacin. PMID- 11480525 TI - Functional MRI activity in the thalamus and occipital cortex of anesthetized dogs induced by monocular and binocular stimulation. AB - The neuroanatomy of the mammalian visual system has received considerable attention through electrophysiological study of cats and non-human primates, and through neuroimaging of humans. Canine neuroanatomy, however, has received much less attention, limiting our understanding of canine vision and visual pathways. As an early step in applying blood oxygenation level dependant (BOLD) functional magnetic resonance imaging (fMRI) for veterinary use, we compared visual activity in the thalamus and occipital cortex of anesthetized dogs presented with binocular and monocular visual stimuli. Activity in the left and right thalamus and occipital cortex during monocular stimulation was also compared. Six beagles were presented with a vertical grating visual stimulus and scanned at 4 Tesla. Each dog was scanned twice under each of 3 anesthetic protocols (isoflurane, propofol, and fentanyl/midazolam). We found: 1) significant BOLD activation in the lateral geniculate nucleus (LGN) of the thalamus and the occipital cortex; 2) a significantly larger area of activation in the LGN during monocular stimulation than during binocular stimulation; and 3) that activity in the hemisphere contralateral to the stimulus was not significantly greater than that ipsilateral to it. PMID- 11480526 TI - Experimental infection of specific pathogen free piglets with French strains of Streptococcus suis capsular type 2. AB - A standardized model of Streptococcus suis type 2 infection in specific-pathogen free piglets, housed in high-security barns, was used to compare the virulence of 3 French field strains of S. suis serotype 2 isolated from tonsils of a healthy pig (strain 65) or from diseased pigs (meningitis, strain 166', or septicemia, strain 24). In one of the 2 trials, 7-week-old pigs, in 3 groups of 8, were inoculated intravenously with 2 x 10(8) colony-forming units of S. suis type 2. In each group, 1 uninfected animal was a sentinel. Eight animals were also used as negative control group. The experiment was repeated under similar conditions with strains 65 and 166'. Virulence differed markedly among these S. suis strains when clinical signs, zootechnical performances, lesions, and bacteriological data were analyzed. Strain 65 did not induce clinical signs in inoculated pigs. In contrast, pigs infected with the other 2 strains exhibited clinical signs and typical lesions of S. suis type 2 infections. Differences in virulence were also observed between the 2 virulent strains. Sentinel animals exhibited the same manifestations as those recorded in inoculated piglets. Results were similar in the second trial, indicating that under the present experimental conditions, results were reproducible. The standardized conditions described in this study could be a useful tool to further study about the S. suis infection. PMID- 11480527 TI - Tightening the screws on human subjects protection. PMID- 11480528 TI - Correlates of physical activity in chronic obstructive pulmonary disease. AB - BACKGROUND: Physical activity is a key dimension of functional status in people with chronic obstructive pulmonary disease (COPD), and the central target of interventions in this group. OBJECTIVES: To determine the relationships among functional performance measured as physical activity, functional capacity, symptom experiences, and health-related quality of life in people with COPD. METHOD: Cross-sectional, descriptive study. Convenience sample of 63 outpatients with COPD studied prior to entry into a pulmonary rehabilitation program. RESULTS: Daily physical activity, as measured by an accelerometer, was strongly associated with maximal distance walked during a 6-minute walk test (r = .60, p < .00), level of airway obstruction (r = .37, p < .01), walking self-efficacy (r = .27, p < .05), and physical health status (r = .40, p < .01). Physical activity was not correlated with self-report of functional status. The only predictor of physical activity was the 6-minute walk test. CONCLUSIONS: Accelerometer measurement of functional performance was most significantly related to walking abilities. This methodology represents a novel approach to measuring an important dimension of functional status not previously well quantified. PMID- 11480529 TI - Supplementation with dietary fiber improves fecal incontinence. AB - BACKGROUND: Human studies have shown that dietary fiber affects stool composition and consistency. Because fecal incontinence has been shown to be exacerbated by liquid stools or diarrhea, management strategies that make stool consistency less loose or liquid may be useful. OBJECTIVE: To compare the effects of a fiber supplement containing psyllium, gum arabic, or a placebo in community-living adults who were incontinent of loose or liquid stools. Mechanisms underlying these effects (e.g., fermentation of the fibers and water-holding capacity of stools) were examined. METHODS: Thirty-nine persons with fecal incontinence of loose or liquid stools prospectively recorded diet intake and stool characteristics and collected their stools for 8 days prior to and at the end of a 31-day fiber supplementation period. During the fiber supplementation period, they ingested psyllium, gum arabic, or a placebo by random assignment. RESULTS: In the baseline period, the groups were comparable on all variables measured. In the fiber supplementation period, (a) the proportion of incontinent stools of the groups ingesting the fiber supplements was less than half that of the group ingesting the placebo, (b) the placebo group had the greatest percentage of stools that were loose/unformed or liquid, and (c) the psyllium group had the highest water-holding capacity of water-insoluble solids and total water-holding capacity. The supplements of dietary fiber appeared to be completely fermented by the subjects as indicated by nonsignificant differences in total fiber, short chain fatty acids and pH in stools among the groups in the baseline or fiber supplementation periods. CONCLUSIONS: Supplementation with dietary fiber from psyllium or gum arabic was associated with a decrease in the percentage of incontinent stools and an improvement of stool consistency. Improvements in fecal incontinence or stool consistency did not appear to be related to unfermented dietary fiber. PMID- 11480530 TI - A comparison of two methods of assessing disease activity in the joints. AB - BACKGROUND: Considerable debate has occurred concerning the utility of different methods of obtaining joint counts and their usefulness in predicting outcomes in persons with rheumatoid arthritis. OBJECTIVE: The purpose of this study was to compare two methods of assessing disease activity in the joints (clinician joint count, self-reported joint count), and to compare their relative utility in predicting two methods of assessing outcomes (self-reported ratings of impairment and pain, objective performance index) with and without controlling for negative affectivity. METHOD: Data for this study were obtained during home visits from 185 persons diagnosed with rheumatoid arthritis. Individuals completed a series of self-report measures including the joint count. Trained research assistants completed a 28-joint count and timed participants on a series of measured performance activities (e.g., grip strength, pinch strength, walk time). RESULTS: The self-report joint count was highly correlated with the clinician joint count and also accounted for as much, if not more, variance in the subjective outcome measures than did clinician assessments. Both types of indicators predicted unique variance in the objective performance index. CONCLUSIONS: For most research purposes, measures such as self-report joint counts have sufficient validity to be used in place of more costly clinician assessment of joint counts. PMID- 11480531 TI - Physical functioning of elderly cancer patients prior to diagnosis and following initial treatment. AB - BACKGROUND AND OBJECTIVES: Using an instrument to measure physical functioning that was normed to the U.S. population, data were obtained from patients with a new diagnosis of breast, colon, lung, and prostate cancer. Two questions were addressed: (a) after controlling for age, and number of comorbid conditions, do site and stage of cancer predict functional limitations prior to diagnosis; (b) using age adjusted national norms on physical functioning, how well do age, number of comorbid conditions, stage, treatment and cluster of symptoms (pain, fatigue, and insomnia) explain changes in physical function between 3 months prior to and 8 weeks following diagnosis? METHODS: Patients 65 years of age and older were accrued from 24 community oncology settings. Consenting patients were interviewed within 8 weeks of initial treatment. The SF-36 was used to measure physical functioning. Comorbidity and symptom experience were assessed through patient report and site and stage of cancer from record audits. RESULTS: Prior to diagnosis of cancer, patients were comparable in physical functioning to the U.S. population aged 55-64, a full decade younger than the sample of cancer patients. Site and stage of disease did not account for variations in physical functioning prior to diagnosis. Compared against national norms, patients with more extensive treatments (surgery plus adjuvant therapy) reported greater loss in functioning. Pain, fatigue, and insomnia had a consistent and significant effect on losses in functioning unrelated to patients' treatments or their comorbid conditions. CONCLUSIONS: Site and stage of cancer prior to diagnosis do not affect functioning. Older cancer patients report higher functioning than their counterparts in the U.S. population. Changes in functioning following diagnosis varied by cancer site. Treatments were related to loss in functioning, but comorbidity was not. Pain, fatigue, and insomnia were significant and independent predictors of change in patient functioning. This underscores the importance of interventions to manage symptoms early in the course of treatment for individuals. PMID- 11480532 TI - Symptom predictors of acute coronary syndromes in younger and older patients. AB - BACKGROUND: Symptoms, a key element in the patient's decision to seek care, are critical to appropriate triage, and influence decisions to pursue further evaluation and initiation of treatment. Although many studies have described symptoms associated with acute coronary syndromes (ACS), few, if any, have examined symptom predictors of ACS and whether they differ by patients' age. OBJECTIVES: To explore symptom predictors of ACS in younger (< 70 years) and older (> or = 70 years) patients. To test the hypothesis that typical symptoms are predictive of ACS in younger patients, but are less predictive in older patients. METHOD: Secondary analysis of observational data gathered on 531 patients presenting to the emergency department of a regional cardiac referral center in New England with symptoms suggestive of ACS. RESULTS: Bivariate analyses revealed no symptoms significantly (p < .01) associated with ACS in older patients. In younger patients presence of chest symptoms and the total number of typical symptoms reported were significantly (p < .01) associated with ACS. After adjustment for age and gender, typical symptoms that were positive predictors of ACS in younger patients included chest symptoms (OR 2.37, 95% CI 1.32-4.27, p = .004) and arm pain (OR 1.78, 95% CI 1.03-3.09, p = .040). Additionally, the total number of typical symptoms reported (OR 1.68, 95% CI 1.31 2.15, p < .001) was a positive predictor of ACS in younger patients. The atypical symptom of fatigue (OR 2.52, 95% CI 1.10-5.81, p = .029) was a significant positive predictor of ACS, whereas dizziness/faintness (OR .50, 95% CI .26-.91, p = .024) was a significant negative predictor of ACS in younger patients. Logistic regression analysis using the entire sample revealed an interaction between age and number of typical symptoms indicating that younger patients had a 36% greater odds for ACS for each additional typical symptom present compared with older patients (OR 1.36, 95% CI 1.02-1.83, p = .038 for interaction between age and number of typical symptoms reported). The model with the interaction between age and chest symptoms revealed a borderline association (p = .10 for the interaction between age and chest symptoms), with younger patients being more likely than older patients to report chest symptoms. CONCLUSIONS: Typical symptoms are predictive of ACS in younger patients and less predictive in older patients. PMID- 11480533 TI - Comparative analysis of the performance of the Postpartum Depression Screening Scale with two other depression instruments. AB - BACKGROUND: Postpartum depression affects approximately 13% of mothers but up to 50% of all cases of this tragic illness can go undetected. OBJECTIVE: The purpose of this study was to compare the performance of a newly created instrument, the Postpartum Depression Screening Scale (PDSS), with the Edinburgh Postnatal Depression Scale (EPDS) and a general depression scale, the Beck Depression Inventory-II (BDI-II). METHOD: In this methodological design a total of 150 new mothers completed these three instruments in random order, followed immediately by a DSM-IV diagnostic interview. Using the LABROC I program, the areas under each of the instrument's Receiver Operator Characteristic (ROC) curves were compared to determine if they were significantly different. RESULTS: Eighteen (12%) of the women were diagnosed with major postpartum depression, 28 women (19%) with minor postpartum depression, and 104 women (69%) with no depression. Compared to the EPDS, the PDSS had a significantly larger area under the ROC curve when screening for major or minor postpartum depression. When using the published recommended cut-off scores for major depression for the three instruments, the PDSS achieved the highest combination of sensitivity, 94%, and specificity, 98%. When detecting women with major or minor postpartum depression, the PDSS again yielded the highest combination of sensitivity (91%) and specificity (72%) of the three instruments. The PDSS identified 17 (94%) of the women diagnosed with major postpartum depression, the EPDS identified 14 of these women (78%), and the BDI-II identified 10 of the 18 women (56%). CONCLUSION: If mothers identified as "most depressed" are substantially determined by the instrument used, the implications for both research and clinical practice are significant. Researchers and clinicians need to be aware of the differential sensitivity of depression instruments which, while supposedly measuring the same construct, are focused on different components of this mood disorder. PMID- 11480534 TI - Rhodes Index of Nausea and Vomiting--Form 2 in pregnant women. A confirmatory factor analysis. AB - BACKGROUND: Despite widespread application of Rhodes Index of Nausea and Vomiting Form 2 (INV2) in practice and research, empirical analyses have not been consistently performed to verify the a priori factors that guided the subclass construction of the symptoms. OBJECTIVES: To examine the dimensional structure of Rhodes INV in a sample of pregnant women. METHOD: Data were collected from 152 pregnant women who were experiencing some degree of nausea and vomiting during early pregnancy and analyzed using structural equation modeling techniques. Five competing measurement structures were tested and compared. The structure (model) that provided the closest fit to the data was selected and relationships (factor loadings) between the constructs and indicators were established. RESULTS: The model fitting the data the closest was a three-factor structure measuring nausea, vomiting, and retching as three separate, but correlated dimensions. The factor loadings were high (0.73-0.96) and significant (p < .001). The model treating nausea and vomiting as a one-factor concept as well as the model including two factors named symptom occurrence and symptom distress did not fit the data. CONCLUSION: Rhodes INV2 is a valid measurement tool if subscales are formed to reflect the multidimensional structure of nausea and vomiting in pregnancy. PMID- 11480535 TI - Evaluation of a single-pass intestinal-perfusion method in rat for the prediction of absorption in man. AB - Prediction of the fraction of dose absorbed from the intestine (Fa) in man is essential in the early drug discovery stage. In-vitro assays in Caco-2 and MDCK cells are routinely used for that purpose, and their predictive value has been reported. However, in-situ techniques might provide a more accurate estimation of Fa. In this study, we evaluated a single-pass intestinal-perfusion (SPIP) method in the rat for its use in the prediction of absorption in man and compared it with a previous report using cell-based assays. Effective permeability coefficients (Peff) were determined in rats for 14 compounds, and ranged from 0.043x 10(-4) cm s(-1) to 1.67 x 10(-4) cm s(-1). These values strongly correlated (r2 = 0.88) with reported Peff values for man. In addition, the Spearman rank correlation coefficient calculated for in-situ-derived Peff and absorption in man was 0.92 while for the previously tested in-vitro Caco-2 and MDCK systems vs absorption in man, the correlation coefficients were 0.61 and 0.59, respectively. SPIP provided a better prediction of human absorption than the cell-based assays. This method, although time consuming, could be used as a secondary test for studying the mechanisms governing the absorption of new compounds, and for predicting more accurately the fraction absorbed in man. PMID- 11480536 TI - Metabolism of sulphobromophthalein I: positional isomers of sulphobromophthalein monoglutathione conjugate. AB - Three positional isomers of sulphobromophthalein glutathione monoconjugate (BSP mGSH) were detected using a paired-ion HPLC method that employs triethylamine phosphate (TEA-H3PO4) as a pairing agent. To confirm that these compounds were glutathione (GSH) conjugates, sulphobromophthalein (BSP) was incubated with a four-fold volume of GSH under alkaline ammonium hydroxide. At least 6 metabolites (3 di-GSH conjugates and 3 isomers of mono-GSH conjugates) were produced under these conditions. The three mono-GSH conjugates were each purified and identified as compounds with a molecular weight of 1,020 according to FAB mass spectrometry results. Positional isomers of BSP-GSH were provisionally distinguished via the addition of the symbols alpha, beta and delta to the end of each abbreviation, to reflect the amount of isomers present. Thus, the isomer present in the largest quantity was termed BSP-mGSH(alpha), the second most abundant isomer was termed BSP-mGSH(beta) and the third was termed BSP-mGSH(delta). Interestingly, a species difference was recognized in that rat cytosol GSH S-transferase (GST) primarily produced BSP-mGSH(alpha), whereas guinea-pig cytosol generated BSP-mGSH(delta), BSP-mGSH(alpha) and BSP-mGSH(beta) equally and rabbit cytosol mainly produced BSP mGSH(beta). PMID- 11480537 TI - Designing multidrug-resistance modulators circumventing the reverse pH gradient in tumours. AB - Multidrug-resistant tumours often exhibit a reverse pH gradient (acid outside), as they have an acid extracellular pH (pHe) and a neutral alkaline intracellular pH (pHi). This study was designed to test the hypothesis that the ability of lipophilic drugs to mediate multidrug resistance (MDR) reversal by interacting with the membrane phospholipids may be correlated with pH in resistant tumours. The permeation properties of five MDR modulators were therefore studied at 37 degrees C by quantifying their ability to induce the leakage of Sulfan blue through unilamellar anionic liposomes, over the range pH 6.5-7.7, and in the absence of any membrane potential (pHe = pHi). The dye leakage induced by two calcium blockers (diltiazem and verapamil) and two antiparasitic agents (thioacridine derivative and mepacrine) was found to significantly increase with the pH of the medium (P < 0.001), whereas that induced by a non-ionic detergent (Triton X-100) showed almost no pH-dependent variations. This process was a cooperative one (0.8 < Hill coefficient < 8.5) and the permeation doses inducing 50% dye leakage (PD50) ranged from 1.6 to 36.0 mM. The permeation ability of the MDR modulators (log(1/PD50)) significantly increased with their octanol-buffer distributions (logD) (slope = 0.35+/-0.06; y intercept = 1.65 +/- 0.14; P < 0.0001) and significantly decreased with their net electric charge (z) (slope = 0.48+/-0.07; y intercept = 2.85+/-0.08; P < 0.0001). A highly significant multiple correlation was found to exist between the variations of log(1/PD50) with those of logD and z (dlog(1/PD50)/dlogD = 0.21 +/- 0.05; dlog(1/PD50)/dz = 0.34+/-0.07; y intercept = 2.27+/-0.17; P < 0.000001). The results provide evidence that in resistant tumours (acid pHe and neutral alkaline pHi), the MDR reversal might be enhanced by favourable drug-membrane interactions if the modulators are designed in the form of highly lipophilic (logP approximately equals 4) mono-basic drugs with a near neutral pKa (pKa approximately equals 7 8). PMID- 11480538 TI - Apoptosis induced by doxorubicin and cinchonine in P388 multidrug-resistant cells. AB - Acquired drug resistance is a major factor in the failure of doxorubicin-based cancer chemotherapy. We determined the ability of cinchonine to reverse doxorubicin drug resistance in a doxorubicin-resistant leukaemia cell line (mouse P388/DOX). A non-cytotoxic concentration of cinchonine (10 microM) increased the sensitivity to doxorubicin of multidrug-resistant P388/DOX cells and significantly enhanced the doxorubicin-induced apoptosis and DNA fragmentation in resistant cells, but had no effect in parent cells. Time-course studies demonstrated that DNA fragmentation was present 24 h after incubation with doxorubicin and cinchonine, indicating that DNA degradation was a preceding event. In cultured cells, cinchonine increased the intracellular accumulation of doxorubicin in the resistant cells in a dose-dependent manner. Using flow cytometry to measure the inhibition of the P-glycoprotein (P-gp) dependent efflux of rhodamine 123, cinchonine was found to be considerably more effective than quinine. The results with cinchonine suggest that there may be quinine derivatives with a similar capacity to inhibit drug transport by P-gp. Additionally, the G2/M phase cell population in resistant cells is increased by doxorubicin/cinchonine treatment. Exposure of resistant cells to 1 microM doxorubicin and 10 microM cinchonine resulted in the expression of Fas (APO 1/CD95) in cells after 6 h. These studies demonstrate that the cell killing effects of doxorubicin and cinchonine in resistant cells PMID- 11480539 TI - Cisapride raises the bioavailability of paracetamol by inhibiting its glucuronidation in man. AB - The effect of cisapride on plasma concentrations of paracetamol was investigated with respect to hepatic metabolism. Paracetamol (1 g) together with cisapride (7.5 mg) or placebo was orally administered to five healthy male volunteers. Venous blood samples were taken before and after administration. Plasma paracetamol and its glucuronide and sulfate conjugates were measured by HPLC. The pharmacokinetic variables were calculated from the plasma concentration-time curves of each volunteer. The area under the plasma paracetamol concentration time curve from 0 to 180 min (mean +/- s.d.) increased from 1875.0 +/- 112.8 micrg min mL(-1) (placebo coadministration) to 2238.8 +/- 125.8 microg min mL(-1) (cisapride coadministration) (P < 0.01). The mean maximum plasma paracetamol concentration(18.2 microg mL(-1))with placebo was reached 30 min after administration, whereas mean maximum plasma paracetamol concentration (21.2 microg mL(-1)) with cisapride occurred 45 min after administration. The plasma paracetamol concentrations with cisapride were significantly greater at 45 to 120 min after administration compared with placebo. Plasma paracetamol glucuronide conjugate concentrations with cisapride were decreased at 15 to 60 min compared with placebo (P< 0.05), whereas plasma paracetamol sulfate conjugate concentrations did not change significantly. Hence the coadministration of paracetamol with cisapride reduced plasma paracetamol glucuronide concentrations and increased plasma paracetamol concentrations, presumably due to inhibition of paracetamol metabolism via paracetamol glucuronyltransferase. Thus, care is necessary when paracetamol and cisapride are coadministered. PMID- 11480540 TI - Synthesis and pharmacological evaluation of 5-dialkylaminomethyl-2-amino-2 oxazolines as H1-antagonists. AB - New 5-dialkylaminomethyl-2-amino-2-oxazolines have been synthezised in two steps from the corresponding dialkylamines. They were evaluated in-vitro as H1 antagonists. Compounds 1c, 1d and 1j significantly antagonized histamine-induced contraction of guinea-pig trachea with a rightward shift of the concentration response curve to histamine. Compound 1f, 5-[(4-benzyl-1-piperidinyl)methyl]-2 amino-2-oxazoline, induced an increase in acetylcholine Emax (the maximal response to acetylcholine 10(-3) M) and a shift to the left of the concentration response curve. The lack of effect of this compound on histamine-induced contraction rules out a non-selective potentiation of the contraction mechanisms. Preliminary structure-activity results were reported partly based on physicochemical results. PMID- 11480541 TI - A flexible approach to the design of new potent substance P receptor ligands. AB - The development of small-molecule antagonists of the substance-P-preferring tachykinin NK1 receptor offers an excellent opportunity to exploit these molecules as novel therapeutic agents in diverse pathologies such as depression, emesis or asthma. GR71251 has previously been identified as a potent and selective substance-P-receptor antagonist. We have therefore undertaken the synthesis of new pseudopeptidic analogues based on the C-terminal sequence of GR71251. The evaluation of binding affinities toward NK1 and NK2 receptors has enabled us to propose new selective NK1 ligands with high affinity. Structure activity relationships showed that the Trp-OBzl(CF3)2 moiety is essential for NK1 affinity and that the introduction of building units such as spirolactam, lactam or proline, leading to a constrained peptide, increased selectivity for NK1 receptors. These compounds constitute a useful starting point for new substance P antagonists and represent an attractive lead series for further studies on the design of specific NK1 antagonists. PMID- 11480542 TI - Antimalarial in-vivo activity of bis(9-amino-6-chloro-2-methoxyacridines). AB - In the fight against malaria, chemotherapy using bisacridines may represent an alternative method to overcoming chloroquine-resistance. Eight bis(9-amino-6 chloro-2-methoxyacridines), in which acridine moieties were linked by polyamines substituted with a side chain, were tested for their in-vivo activity upon mice infected by Plasmodium berghei. Three of the compounds revealed antimalarial activity but no relationship could be deduced from a comparison of in-vitro and in-vivo activities. N-alkylation of the central amino group generated toxicity and, therefore, only compounds N-acylated in this position can be selected as leads. PMID- 11480543 TI - Spirohydantoins from D-ribose as new potent enzymatic inhibitors. AB - Anomeric spirohydantoin derivatives from monosaccharides are known for various biological properties. We describe herein the synthesis of the 3-spirohydantoin derivatives of D-allose and D-ribose. The key step is the stereoselective glyco alpha-aminonitrile formation from ulose derivatives of D-glucose and D-xylose using titanium tetra-isopropoxide as a mild and efficient catalyst. Target compounds were synthesized from these intermediates. The glucidic moiety was partially or totally deprotected under acidic conditions. These new heterocyclic monosaccharidic derivatives are potent glycogen phosphorylase inhibitors. PMID- 11480544 TI - Asymmetric synthesis of novel isoindolines: azasaccharide mimics as potential enzyme inhibitors. AB - 2',3'-Dideoxy-2',3'-didehydronucleosides and azasaccharides are known to possess antiviral activity. The synthesized 1-methoxyisoindoline system (10), which is related to the above nucleosides, is potentially stable in-vivo. The 1 methoxyisoindoline was synthesized from the achiral phthalaldehyde in 10 steps via an enantiomerically pure diol obtained by Sharpless asymmetric dihydroxylation. The new heterocyclic compound is an azosaccharide mimic which provides an access to a new series of nucleoside analogues with potential as antiretroviral agents (anti-HIV) and as glycosidase inhibitors. PMID- 11480545 TI - Conformational analysis of tripeptide Ac-Lys-Pro-Val-NH2, COOH-terminal sequence of alpha-MSH. AB - Alpha-melanocyte stimulating hormone (alpha-MSH) is an endogenous linear tridecapeptide which interacts with the melanocortin receptors (MC1-R to MC5-R) to mediate its biological effects. Antipyretic and anti-inflammatory activities of alpha-MSH are due to the COOH-terminal peptide sequence, Lys-Pro-Val (alpha MSH[11-13]). This tripeptide might be useful as a therapeutic agent in the control of fever and inflammatory reactions. With this aim, a theoretical conformational study of the tripeptide has been carried out using molecular dynamics. The obtained conformational space has been classified into families according to the letter-code convention to partition the phi-psi map. The lowest energy conformations of each family were used as templates to design six models of conformationally constrained nonpeptide analogues. PMID- 11480546 TI - Studies on the reactivity of a tertiary allylic alcohol in an acetophenonic series, a model for natural products synthesis. AB - The synthesis of benzopyranic simplified analogues of dibenzopyranic natural compounds is described, together with the access to a precursor of a new furobenzopyranic natural product. These natural products have anti-cancer activity. The 1,3-diacetoxy-2-acetyl-4-(3-hydroxy-3-methylbut-1-enyl)benzene synthone is used as a common precursor to these structures. PMID- 11480547 TI - 5-substituted 3,4-dihydro-3-amino-2H-1-benzopyran derivatives: synthesis and interaction with serotoninergic receptors. AB - A new series of 3,4-dihydro-3-amino-2H-1-benzopyran derivatives (1 and 2) bearing various substituents on the 5-position was successfully prepared via palladium mediated cross-coupling reactions. Some of the new compounds showed high affinity for 5-HT1A and 5-HT7 receptors. The best affinity for the 5-HT1A and 5-HT7 receptors was obtained for 2b (Ki = 0.3 nM for 5-HT1A and 3.1 nM for 5-HT7). The anxiolytic activity of compound 2b was evaluated. PMID- 11480548 TI - Solid phase synthesis of sulphonamides: novel ligands of 5-HT6 receptors. AB - Using solid phase synthesis techniques, we have rapidly obtained a series of eight aryl sulphonamides derived from putrescine. These conjugates with various aryl groups were evaluated for their affinity towards 5-HT6 receptors in man. This evaluation revealed the interest of two compounds which present the same activity level, in the submicromolar range, as two reference derivatives. The most potent will be considered as a new lead for further investigations. PMID- 11480549 TI - Effect on insulin release of compounds structurally related to the potassium channel opener 7-chloro-3-isopropylamino-4H-1,2,4-benzothiadiazine 1,1-dioxide (BPDZ 73): introduction of heteroatoms on the 3-alkylamino side chain of the benzothiadiazine 1,1-dioxide ring. AB - 7-Chloro-3-pyridyl(alkyl)amino-4H-1,2,4-benzothiadiazine 1,1-dioxides and 3 alkylamino-7-chloro-4H-1,2,4-benzothiadiazine 1,1-dioxides containing one or more heteroatoms on the side chain in the 3 position have been synthesized in an attempt to discover new potent KATP-channel openers. The compounds were tested as putative pancreatic B-cells KATP channel openers by measuring their inhibitory activity on the insulin releasing process. The influence on the biological activity of the nature of the side chain in the 3 position is discussed. PMID- 11480550 TI - Synthesis of diverse 4,5-dihydro-3(2H)-pyridazinones on Wang resin. AB - An efficient solid-phase synthesis of structurally diverse 4,5-dihydro-3(2H) pyridazinones is described using readily available substituted 4-oxo-butanoic acids. Polymer-supported gamma-keto-esters prepared from Wang resin reacted with several hydrazines to afford the corresponding hydrazones. A protocol developed in mild conditions without isolating the intermediate hydrazone led to pyridazinones in good yields after a cyclization cleavage approach. This successful strategy represents an attractive method for a rapid synthesis of heterocyclic libraries for biological evaluation. PMID- 11480551 TI - Contradistinction between doxorubicin and epirubicin: in-vivo metabolism, pharmacokinetics and toxicodynamics after single- and multiple-dosing in rats. AB - There is compelling in-vitro evidence that the evaluation of doxorubicin or epirubicin pharmacokinetics based solely on plasma concentration may not fully elucidate the differences between the two drugs. Both compounds bind to erythrocytes and their different binding to haemoglobin may influence their disposition in the body. The purpose of the present study was to compare the pharmacokinetics and metabolism of doxorubicin and epirubicin based on the plasma concentration, amount associated with blood cells and simultaneous monitoring of biliary and urinary elimination of unchanged drug and metabolites after single- and multiple-dose injections. The level of sarcoplasmic reticulum Ca2+ATPase in the heart was also measured as a biomarker of cardiotoxicity. Male Sprague-Dawley rats were treated in a parallel design with doxorubicin or epirubicin on a multiple-dosing basis (4 mg kg(-1) per week) or as a single dose injection (20 mg kg(-1)). Blood, urine and bile samples were collected periodically after each dose in the multiple-dosing regimen and the single dose injection, and at the end of each experiment the hearts were removed. The concentrations of each drug in plasma, blood cells, bile and urine samples were determined, and by simultaneous curve-fitting of plasma and bile data according to compartmental analysis, the pharmacokinetic parameters and constants were estimated. The concentration of drug associated with blood cells was analysed according to non-compartmental analysis. The bile and urine samples provided the in-vivo metabolic data. The level of Ca2+ATPase in the heart, determined by Western blotting, was used as the toxicodynamic parameterto correlate with the kinetic data. Multiple-dosing regimens reduced the total plasma clearance and increased the area under the plasma concentration-time curve of both drugs. Also, the area under the curve of doxorubicin associated with blood cells increased with the weekly doses, and the related mean residence time (MRT) and apparent volume of distribution (Vdss) were steadily reduced. In contrast to doxorubicin, the MRT and Vdss of epirubicin increased significantly. Metabolic data indicated significant differences in the level of alcohol and aglycones metabolites. Doxorubicinol and doxorubicin aglycones were significantly greater than epirubicinol and epirubicin aglycone, whereas epirubicinol aglycone was greater than doxorubicinol aglycone. The area under the blood cells concentration-time curve correlated linearly with the changes in Ca2+ATPase net intensity. The results of this study demonstrate the importance of the kinetics of epirubicin and doxorubicin associated with blood cells. Linear correlation between the reduction of net intensity of the biomarker with the area under the curve of doxorubicin associated with blood cells confirms that the differences between the two compounds are related to their interaction with blood cells. This observation together with the observed differences in metabolism may underline a significant role for blood cells in distribution and metabolism of doxorubicin and epirubicin. PMID- 11480552 TI - Biostability and pharmacokinetics of LJP 920, an octameric Gal (alpha1-3) Gal conjugate for the inhibition of xenotransplantation rejection. AB - Antibodies to an alpha-galactosyl saccharide structure present in human serum are associated with hyperacute rejection and delayed xenograft rejection after pig-to primate xenotransplantation. To overcome this major barrier to the xenotransplantation, LJP 920, a galactosyl alpha1-3 galactose (Gal (alpha1-3) Gal) coupled to a non-immunogenic platform at a valency of eight Gal (alpha1-3) Gal molecules/platform, was synthesized to clear circulating antibodies and to inhibit their production by B cells that produce these antibodies. Herein we report on the stability of UP 920 in biological media and its pharmacokinetic profile. Incubation of LJP 920 with mouse serum or liver microsomes at 37 degrees C for 2 days showed no indication of degradation of the conjugate as detected by a reversed-phase HPLC method, indicating that the conjugate is not subject to enzymatic metabolism. After intravenous administration of LJP 920 to mice at the doses of 20 and 100 mg kg(-1), UP 920 serum concentration decreased rapidly, showing a biphasic pattern, with a distribution half-life of 3 min and an elimination half-life of more than 30 min, respectively. The serum-to-erythrocyte concentration ratio of UP 920 was 33- and 36-fold excess at 0.5 and 5 min, respectively, after intravenous administration (100 mg kg(-1)). Both Cmax and AUC values increased in a dose-proportional manner. UP 920 displayed a great distribution to well-perfused tissues. It was eliminated mainly through renal excretion in the unchanged form, which accounted for 23% of the total amount within 8 h of dosing. PMID- 11480553 TI - What's in a name...apropos MPO. PMID- 11480554 TI - The cellular pathogenesis of paroxysmal nocturnal haemoglobinuria. AB - Paroxysmal nocturnal haemoglobinuria (PNH) is a unique disorder characterised by the triad of intravascular haemolysis, thrombosis and bone marrow failure. In the early seventies it was shown that PNH is a clonal disease; and in the nineties the molecular basis of the PNH abnormality was elucidated. However, what makes a PNH clone expand is still not known. Here, we suggest that this is due to somatic cell selection, resulting from the presence in the patient of autoreactive T cells that target glycosylphosphatidylinositol (GPI) in the context of an MHC like molecule on the surface of haemopoietic stem cells. PNH cells would escape damage precisely because they have lost most or all of their ability to produce GPI. PMID- 11480555 TI - Ceramide regulates cellular homeostasis via diverse stress signaling pathways. AB - The sphingolipid ceramide is an important second signal molecule that regulates diverse signaling pathways involving apoptosis, cell senescence, the cell cycle, and differentiation. For the most part, ceramide's effects are antagonistic to growth and survival. Interestingly, ceramide and the pro-growth agonist, diacylglycerol (DAG) appear to be regulated simultaneously but in opposite directions in the sphingomyelin cycle. While ceramide stimulates signal transduction pathways that are associated with cell death or at least are inhibitory to cell growth (eg stress-activated protein kinase, SAPK, pathways), DAG activates the classical and novel isoforms of the protein kinase C (PKC) family. These PKC isoforms are associated with cell growth and cell survival. Furthermore, DAG activation of PKC stimulates other signal transduction pathways that support cell proliferation (eg mitogen-activated protein kinase, MAPK, pathways). Thus, ceramide and DAG generation may serve to monitor cellular homeostasis by inducing pro-death or pro-growth pathways, respectively. The production of ceramide is emerging as a fixture of programmed cell death. Ceramide levels are elevated in response to diverse stress challenges including chemotherapeutic drug treatment, irradiation, or treatment with pro-death ligands such as tumor necrosis factor alpha, TNF alpha. Consistent with this notion, ceramide itself is a potent apoptogenic agent. Ceramide activates stress activated protein kinases like c-jun N-terminal kinase (JNK) and thus affects transcription pathways involving c-jun. Ceramide activates protein phosphatases such as protein phosphatase 1 (PP1) and protein phosphatase 2 (PP2A). Ceramide activation of protein phosphatases has been shown to promote inactivation of a number of pro-growth cellular regulators including the kinases PKC alpha and Akt, Bcl2 and the retinoblastoma protein. A new role has recently emerged for ceramide in the regulation of protein synthesis. Ceramide-induced activation of double stranded RNA-dependent protein kinase (PKR), a protein kinase important in anti viral host defense mechanisms and recently implicated in cellular stress pathways, results in the inhibition of protein synthesis as a prelude to cell death. Taken together, these properties of ceramide suggest that this important second-signal molecule may have useful properties as an anti-neoplastic agent. Thus, strategies to promote ceramide metabolism or use of ceramide analogs directly may one day become useful in the treatment of diseases like leukemia. PMID- 11480556 TI - CD56 antigenic expression in acute myeloid leukemia identifies patients with poor clinical prognosis. AB - CD56 antigen, a 200-220 kDa cell surface glycoprotein, identified as an isoform of the neural adhesion molecules (NCAM), has been found frequently expressed in several lympho-hematopoietic neoplasms including acute myeloid leukemias (AML). In fact, in these latter diseases it has been reported that the presence of CD56 antigen on the blasts of AML patients with t(8;21) (q22;q22), and in those with M3 subtype, identifies a subgroup of patients with a more unfavorable prognosis. On the basis of these findings, we evaluated in 152 newly diagnosed AML patients CD56 surface expression, and results were correlated with morphology, immunophenotype, cytogenetic pattern and clinical outcome. CD56 antigen was recorded in 37 out of 152 cases (24%) and particularly in those with M2 and M5 cytotypes. Moreover, CD56 expression was significantly associated with P glycoprotein (PGP) hyperexpression (P = 0.007), unfavorable cytogenetic abnormalities (P = 0.008) and with a reduced probability of achieving complete remission (CR) (36% vs 68%) (P = 0.035) as well as with a shorter survival (6 vs 12 months) (P = 0.032). In conclusion, CD56 antigenic expression on AML cells represents an important adverse prognostic factor and therefore its presence should be regularly investigated for a better prognostic assessment of AML patients at diagnosis. PMID- 11480557 TI - Plasma hepatocyte growth factor is a prognostic factor in patients with acute myeloid leukemia but not in patients with myelodysplastic syndrome. AB - Hepatocyte growth factor (HGF) is a potent angiogenic factor. The aim of our study was to evaluate plasma HGF levels and their prognostic significance in patients with newly diagnosed acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). The sandwich enzyme immunoassay technique was used to quantify HGF in stored samples obtained before treatment from patients with AML (59 patients) and MDS (42 patients) treated at The University of Texas MD Anderson Cancer Center. HGF levels were significantly higher in patients with AML or MDS than in healthy individuals (P < 0.0001). Higher HGF levels in both AML and MDS correlated significantly with white blood cell (P = 0.000001 for both groups) and monocyte counts (P = 0.0004 and 0.003, respectively), and with poor performance status (P = 0.03 and 0.001, respectively). Using Cox proportional hazard model and HGF levels as a continuous variable, plasma levels of HGF correlated with shorter survival of AML (P = 0.001), but not MDS (P = 0.34) patients. No significant correlation was observed between HGF levels and complete remission rate or duration. In the multivariate analysis HGF retained its significance as prognostic factor in AML (P = 0.02), along with age (P = 0.0005). PMID- 11480558 TI - A phase II study of sequential combination chemotherapy with cyclophosphamide, prednisone, and 2-chlorodeoxyadenosine in previously untreated patients with chronic lymphocytic leukemia. AB - In an earlier study of previously untreated patients with chronic lymphocytic leukemia (CLL), we used a concomitant combination of chlorambucil and 2 chlorodeoxyadenosine and reported overall (OR) and complete (CR) remission rates of 80% and 20%, respectively. After a median follow-up of 5 years, more than 80% of the responders have had a relapse. In the current phase II study of 27 previously untreated patients with CLL, we used a sequential combination of six cycles of intravenous cyclophosphamide (1 g/m2) plus oral prednisone (100 mg/m2 per day for 5 days) followed by two to six cycles of 2-chlorodeoxyadenosine (5 mg/m2 per day for 5 days). The OR and CR rates were 96% and 33%, respectively. After a median follow-up of 29 months, 35% of the responders have had a relapse. Progression-free survival was significantly better in CR patients than in those with partial remission. However, minimal residual disease was phenotypically detected in four of the nine CR patients. Despite the fact that the current OR and CR rates are superior to those seen in a historical cohort treated with a concomitant schedule, a longer follow-up period is needed to assess the durability of these remissions, and a controlled trial is necessary to estimate the impact on overall survival and toxicity. PMID- 11480559 TI - A novel differentiation-inducing therapy for acute promyelocytic leukemia with a combination of arsenic trioxide and GM-CSF. AB - Arsenic trioxide (As2O3) effectively induces clinical remission via apoptosis in relapsed acute promyelocytic leukemia (APL). However, because this new anti leukemic drug is also considered to be a poison, its possible adverse effects are a highly important issue related to its clinical use. We here investigated, both in vitro and in vivo, the effects of a combination of As2O3 and GM-CSF as a novel therapeutic approach for the treatment of APL. Treatment of both retinoic acid (RA)-sensitive and -resistant APL cell lines (NB4 and UF-1 cells, respectively), as well as primary APL cells with a combination of As2O3 and GM-CSF for 4 days resulted in inducing differentiation, but not apoptosis, to mature granulocytes. In addition, a combination of both agents induced degradation of the PML/RARalpha protein. GM-CSF was found to be associated with increased tyrosine phosphorylation of Jak2 kinase in both NB4 and UF-1 cells, and a specific inhibitor of Jak2, AG490, completely blocked the ability of GM-CSF to prevent apoptosis and induce differentiation of As2O3-treated UF-1 cells. In in vivo analysis, As2O3 induced differentiation of APL cells in a RA-resistant APL model of human GM-CSF-producing transgenic SCID mice that had a high level of human GM CSF in their sera. In contrast, As2O3 alone diminished tumors in UF-1 cells transplanted into NOD/SCID mice via induction of apoptosis. In conclusion, a combination of As2O3 and GM-CSF appears to be a novel differentiation-inducing therapy in patients with APL, including relapsed or RA-resistant cases. PMID- 11480560 TI - BIOMED-I concerted action report: flow cytometric immunophenotyping of precursor B-ALL with standardized triple-stainings. BIOMED-1 Concerted Action Investigation of Minimal Residual Disease in Acute Leukemia: International Standardization and Clinical Evaluation. AB - The flow cytometric detection of minimal residual disease (MRD) in precursor-B acute lymphoblastic leukemias (precursor-B-ALL) mainly relies on the identification of minor leukemic cell populations that can be discriminated from their normal counterparts on the basis of phenotypic aberrancies observed at diagnosis. This technique is not very complex, but discordancies are frequently observed between laboratories, due to the lack of standardized methodological procedures and technical conditions. To develop standardized flow cytometric techniques for MRD detection, a European BIOMED-1 Concerted Action was initiated with the participation of laboratories from six different countries. The goal of this concerted action was to define aberrant phenotypic profiles in a series of 264 consecutive de novo precursor-B-ALL cases, systematically studied with one to five triple-labelings (TdT/CD10/CD19, CD10/CD20/CD19, CD34/CD38/CD19, CD34/CD22/CD19 and CD19/CD34/CD45) using common flow cytometric protocols in all participating laboratories. The use of four or five triple-stainings allowed the identification of aberrant phenotypes in virtually all cases tested (127 out of 130, 98%). These phenotypic aberrancies could be identified in at least two and often three triple-labelings per case. When the analysis was based on two or three triple-stainings, lower incidences of aberrancies were identified (75% and 81% of cases, respectively) that could be detected in one and sometimes two triple-stainings per case. The most informative triple staining was the TdT/CD10/CD19 combination, which enabled the identification of aberrancies in 78% of cases. The frequencies of phenotypic aberrations detected with the other four triple-stainings were 64% for CD10/CD20/CD19, 56% for CD34/CD38/CD19, 46% for CD34/CD22/CD19, and 22% for CD19/CD34/CD45. In addition, cross-lineage antigen expression was detected in 45% of cases, mainly coexpression of the myeloid antigens CD13 and/or CD33 (40%). Parallel flow cytometric studies in different laboratories finally resulted in highly concordant results (>90%) for all five antibody combinations, indicating the high reproducibility of our approach. In conclusion, the technique presented here with triple-labelings forms an excellent basis for standardized flow cytometric MRD studies in multicenter international treatment protocols for precursor-B-ALL patients. PMID- 11480561 TI - Chromosomal instability in chromosome band 12p13: multiple breaks leading to complex rearrangements including cytogenetically undetectable sub-clones. AB - During fluorescence in situ hybridization (FISH) analysis of metaphase cells from 70 patients with lymphoid and myeloid hematologic malignancies and chromosomal rearrangements involving band 12p13, we identified nine patients (four with lymphoid malignancies, four with myeloid malignancies and one with biphenotypic leukemia) who showed more complicated rearrangements than we had expected from conventional cytogenetic study. In six patients, multiple breaks occurred in small segments of 12p with subsequent translocations and insertions of these segments into other chromosomes, sometimes to unexpected regions. In three patients additional chromosome breaks resulted in a sub-clone which was cytogenetically indistinguishable from the main clone in each patient based on the cytogenetic analysis. These subtle molecular events were detected exclusively in a region covering TEL/ETV6 and KIP1/CDKN1B. Seven of nine had a previous history of chemo/radiotherapy; all the patients showed complex karyotypes, even though they were newly diagnosed with leukemia. Survival data were available in five patients, and all survived less than 6 months. These findings suggest that the 12p13 region, especially the above-mentioned region, is genetically unstable and fragile. It is likely that multiple chromosome breaks were induced through mutagens used in chemo/ radiotherapy, and are associated with a sub-group of patients with an extremely bad prognosis. PMID- 11480562 TI - Enhanced ability of daniplestim and myelopoietin-1 to suppress apoptosis in human hematopoietic cells. AB - Modified and chimeric cytokines have been developed to aid in the recovery of hematopoietic precursor cells after myeloablative chemotherapy. The interleukin-3 (IL-3) receptor agonist, daniplestim, binds to the IL-3 receptor-alpha subunit with 60-fold greater affinity and induces cell proliferation and colony-forming unit formation 10- to 22-fold better than native IL-3. A chimeric cytokine, myelopoietin-1, composed of daniplestim and a G-CSF receptor agonist binds both the IL-3 and G-CSF receptors. While the in vivo effects of daniplestim and myelopoietin-1 are well described, the mechanisms by which they stimulate growth are not well understood. We have investigated the effects of daniplestim and myelopoietin-1 on the prevention of apoptosis in two human hematopoietic cell lines, OCI-AML.5 and AML 193. Daniplestim and myelopoietin-1 prevented apoptosis to a greater degree than native recombinant IL-3 or G-CSF as determined by annexin V/propidium iodide binding and TUNEL assays. Daniplestim and myelopoietin 1 promoted the maintenance of the mitochondrial membrane potential better than native IL-3 or G-CSF. These cytokines promoted a lower redox potential as higher levels of free radicals were detected after cytokine treatment than in cytokine deprived cells implying increased respiration. These results indicate that daniplestim and myelopoietin-1 are able to prevent apoptosis in hematopoietic cells more effectively than native IL-3 and G-CSF. These effects of daniplestim and myelopoietin-1 may contribute to their effective ability to repopulate hematopoietic precursor cells after chemotherapy. PMID- 11480563 TI - A new matrix metalloproteinase inhibitor SI-27 induces apoptosis in several human myeloid leukemia cell lines and enhances sensitivity to TNF alpha-induced apoptosis. AB - MMP inhibitors are used clinically for the stabilization of tumor growth, thus prolonging survival in cancer patients. However, their role in the treatment of hematopoietic malignancies remains unclear. In the present study, we investigated the effects of a new MMP inhibitor, SI-27, in hematopoietic malignancies. SI-27 alone induces apoptosis in several human myeloid leukemia cell lines such as U937, NB4, and HL60 cells by activating caspase 8, 9, and 3. Apoptosis was measured with annexin V positive staining, a drop in mitochondrial transmembrane potential (deltapsim), presence of hypodiploid DNA, and cleavage of PARP and IkappaBalpha. Furthermore, at lowered concentrations, which did not directly induce apoptosis, SI-27 acted to sensitize U937 cells and other cells to tumor necrosis factor alpha (TNF-alpha)-mediated apoptosis. The accumulation of membrane Fas, the Fas ligand, and TNFR1 were not apparent due to exposure to SI 27, and antagonistic anti-Fas or anti-Fas ligand antibodies did not block SI-27 induced apoptosis. Thus, SI-27-induced apoptosis is not mediated by the Fas pathway. These results suggest that MMP inhibitors, alone or in combination with other cytotoxic agents, can provide a unique method for treating acute myeloid leukemia, refractory to classical anti-cancer drugs, and may thus suppress recurrence. PMID- 11480564 TI - Comparison of the effects of recombinant adenovirus-mediated expression of wild type p53 and p27Kip1 on cell cycle and apoptosis in SUDHL-1 cells derived from anaplastic large cell lymphoma. AB - Recombinant adenoviruses expressing wild-type p53 (AdWTp53) and p27KiP1 (Adp27) were used to compare the effects on cell cycle and apoptosis in SUDHL-1 cells derived from human anaplastic large cell lymphoma. Cells infected with AdWTp53 and Adp27 showed high level of wild-type p53 and p27KiP1 expression, respectively. The expression of these proteins resulted in G1 arrest after 24 h of infection. Although the cells persisted in G1 arrest in both cell populations after 48 and 72 h of infection, the level of apoptosis assessed by TUNEL analysis was higher in cells infected with AdWTp53. Interestingly, apoptosis was more pronounced in cells infected with Adp27 after the initial 24 h and reached a steady state at 48 and 72 h. A lower MOI of Adp27 resulted in G1 arrest associated with a low level of apoptosis in SUDHL-1 cells after 48 h of infection. This was correlated with lower expression of p27KiP1. We postulate that the time-lag and the different level of apoptosis occurring in SUDHL-1 cells infected with AdWTp53 and Adp27 are clearly related to the intrinsic biochemical pathways solicited. In this context our study provides a model to investigate these pathways and better understand the biology of this particular lymphoma. Our data also support a potential application of Adp27 for gene therapy of this lymphoma similarly to AdWTp53 as previously shown. PMID- 11480565 TI - Cell adhesion-mediated drug resistance (CAM-DR) protects the K562 chronic myelogenous leukemia cell line from apoptosis induced by BCR/ABL inhibition, cytotoxic drugs, and gamma-irradiation. AB - Integrin-mediated cellular adhesion to extracellular matrix (ECM) components is an important determinant of chemotherapeutic response of human myeloma cells. Here, we demonstrate that when K562 chronic myelogenous leukemia (CML) cells are adhered to fibronectin (FN), they become resistant to apoptosis induced by the BCR/ABL inhibitors AG957 and STI-571, as well as DNA damaging agents and gamma irradiation. This phenomenon, termed cell adhesion-mediated drug resistance (CAM DR), was induced by adhesion through the alpha5beta1 (VLA-5) integrin. Phosphotyrosine analysis demonstrates that anti-apoptotic signaling through integrins in K562 cells is independent of the tyrosine kinases activated by BCR/ABL, with the possible exception of an unknown 80 kDa protein. Cytoprotection of FN-adhered CML cells indicates that tumor-ECM interactions may be critical for the emergence of drug-resistant tumor populations and treatment failure in this disease. Antagonists of beta1 integrin-mediated adhesion or corresponding signal transduction elements may sensitize CML cells to chemotherapy and prevent resistance to the novel BCR/ABL kinase inhibitors being used for the treatment of this disease. PMID- 11480566 TI - The pharmacodynamic effect of busulfan in the P39 myeloid cell line in vitro. AB - Busulfan is an alkylating agent currently used in the myeloablative conditioning regimen before stem cell transplantation. Its mechanism of action is not fully understood, nor the reason for its narrow therapeutic window. We studied the pharmacodynamics of busulfan in an in vitro cell line model, allowing us to evaluate the effects of various doses and exposure times on clonogeneic capacity, proliferation and apoptosis. Cells were incubated with busulfan in concentrations ranging from 10 to 100 microg/ml for 2, 4 or 8 h, then washed and cultured in busulfan-free medium for 72 h. Area under the concentration-time curve (AUC) was estimated by using the trapezoidal rule from different concentrations and times of incubation. In all assays busulfan affected the cells in an AUC-dependent manner. Induced changes in the biological parameters studied appeared at different time points after exposure to busulfan stopped. Thus, the decrease in proliferation and clonogenic capacity preceded cell cycle arrest in G2 phase and development of apoptosis, implying that apoptosis is a secondary event to interruption of vital metabolic processes. Biochemically, apoptotic changes were typical for chemotherapy-induced apoptosis with caspase activation, cleavage of Bcl-2 and PARP proteins, while cleavage of actin was not observed. Cells were rescued from apoptosis with a general caspase inhibitor ZVAD-fmk, but not with granulocyte colony-stimulating factor (G-CSF). Our results add new information about busulfan pharmacodynamics and mechanisms underlying the cytotoxic effect of the drug. PMID- 11480567 TI - The multi-organ origin of interleukin-5 in the mouse. AB - Murine Ba/F3 cells were transfected with cDNA for the alpha-chain of the murine interleukin-5 (IL-5) receptor and cloned lines of these cells were able to proliferate in response to as little as 2.5 pg/ml of IL-5. The bioassay was demonstrated to be specific for IL-5 and was able to measure IL-5 produced in culture by organs from adult C57BL/6 and BALB/c mice. The highest levels of IL-5 were produced by lung tissue but thymus and bladder consistently produced IL-5 and more variable production was observed by the heart, spleen, muscle, bone shaft, uterus and testes. Bone marrow cells produced no detectable IL-5. Observed levels of production of IL-5 were similar when using organs from mice lacking high-affinity receptors for IL-5 and from nu/nu, RAG-1-/- and NOD/SCID mice lacking T lymphocytes. In inflammatory peritoneal exudates induced by the injection of casein plus bacteria, levels of induced IL-5 were higher if the mice lacked high-affinity receptors for IL-5. The data indicate that T lymphocytes are not the dominant cellular source of IL-5 in organ-conditioned media and that local IL-5 production can occur with a wide range of normal murine organs. PMID- 11480568 TI - Clinicopathologic characteristics of leukemia in Japanese children and young adults. AB - The aim of this study is to clarify the clinicopathologic characteristics of adolescent leukemia in Japan by retrospective analysis. Patients with acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), and myelodysplastic syndrome (MDS), consecutively diagnosed from 1986 to 1999, were enrolled. A total of 3,856 patients from 1 to 15 years of age and 1,803 patients from 15 to 29 years of age were eligible for this study. Demographically, the frequency of AML found was almost constant during the teenage years, whereas the frequency of ALL gradually decreased. The relative frequency of CML and MDS apparently started to increase in patients in their late teens. The relative frequency of M3 and t(15;17) gradually increased during adolescence. Among patients aged 1 to 4 years, M7 was the most frequent FAB subtype. Among patients aged 5 to 9 years, M2 and t(8;21) was the most frequent subtype. The percentage of T cell ALL increased in patients 5 to 9 years old, reaching 31.2% in the 20- to 24-year-old age group. The percentage of patients with hyperdiploidy over 50 chromosomes was highest (17.0%) in patients aged 1 to 4 and decreased to 3.9% in the older teens. The percentage of patients with the Ph1 chromosome increased from 9.9% in teens to 30.0% in patients in their late twenties. When comparing event-free survival (EFS) rates for ALL according to age, the estimated 7-year EFS rate was highest for patients aged 1 to 9 years (65.9%) and intermediate for patients aged 10 to 15 years (48.4%). However, the EFS rate was significantly worse for patients aged 15 to 19 years (19.4%) and 20 to 29 years (17.0%) (P = 0.024). On the other hand, the EFS rate for AML decreased with increasing age, although without statistical significance. The overall survival rates are approximate among all age groups. The results of the study indicate that there are considerable variations in biologic features of leukemia between children and young adults. The prognosis for adolescent leukemia may be improved by introducing pediatric trials, which take into account the prognostic biological features. PMID- 11480569 TI - A novel recurrent translocation t(11;14)(p11;q32) in splenic marginal zone B cell lymphoma. AB - A novel recurrent translocation t(11;14)(p11;q32) was found in three patients with splenic marginal zone B cell lymphoma (MZBCL). Fluorescence in situ hybridization (FISH) studies with IgH probes revealed in all cases involvement of the IgH locus, with breakpoint downstream of the IGVH sequences. Partner genes at 11p11 were not identified. The translocation defined the stem line in two patients, who carried additional cytogenetic aberrations, including a 17p deletion, present in both cases. In one patient a 7q- chromosome was the primary cytogenetic defect, the t(11;14) having been found in four out of 11 abnormal metaphase cells at the time of transformation into high-grade MZBCL. Hematological features in all cases included splenomegaly with peripheral blood (PB) involvement by a monoclonal B cell population consisting of lymphocytes with villous projections and several blast-like cells. The immunophenotype was CD19+; CD22bright+; CD23-, CD10-, CD5-, surface Igbright+. A bone biopsy in one patient revealed an interstitial infiltration with an intrasinusoidal pattern of growth. Histological studies on spleen specimens in two patients showed an expanded marginal zone, with small lymphocytes and several blast-like cells. One patient had a therapy-demanding disease, with partial, short-term responses to cytotoxic treatment; one patient transformed into a high-grade MZBCL involving the gut, the PB and the bone marrow 2 years after diagnosis; one patient was unresponsive to cytotoxic treatment and underwent splenectomy. The t(11;14)(p11;q32) may define a subset of splenic MZBCL with a high-grade component and a relatively aggressive clinical behavior. PMID- 11480570 TI - Frequent demonstration of human herpesvirus 8 (HHV-8) in bone marrow biopsy samples from Turkish patients with multiple myeloma (MM). AB - In order to investigate the frequency of HHV-8 in MM patients from another geographic location, we obtained fresh bone marrow (BM) biopsies from Turkish patients with MM (n = 21), monoclonal gammopathy of undetermined significance (MGUS) (n = 2), plasmacytoma (n = 1) with BM plasma cell infiltration, various hematological disorders (n = 6), and five healthy Turkish controls. The frequency of HHV-8 was analyzed by polymerase chain reaction (PCR) in two independent laboratories in the USA and in Turkey. Using fresh BM biopsies, 17/21 MM patients were positive for HHV-8 whereas all five healthy controls, and six patients with other hematological disorders were negative. Two patients with MGUS, and one patient with a solitary plasmacytoma were also negative. The data from the two laboratories were completely concordant. Also using primer pairs for v IRF and v IL-8R confirmed the results observed with the KS330233 primers. Furthermore, sequence analysis demonstrated a C3 strain pattern in the ORF26 region which was also found in MM patients from the US. Thus, HHV-8 is present in the majority of Turkish MM patients, and the absence of the virus in healthy controls further supports its role in the pathogenesis of MM. PMID- 11480571 TI - Thalidomide for previously untreated indolent or smoldering multiple myeloma. AB - We conducted a clinical trial of thalidomide as initial therapy for asymptomatic smoldering (SMM) or indolent multiple myeloma (IMM). Sixteen patients were studied. Thalidomide was given orally at a dose of 200 mg/day for 2 weeks, and then increased as tolerated by 200 mg/day every 2 weeks to a maximum dose of 800 mg/day. Bone marrow microvessel density (MVD) and angiogenesis grading were estimated using CD34 immunostaining. Six patients had a confirmed response to therapy with at least 50% or greater reduction in serum and urine monoclonal (M) protein. When minor responses (25-49%) decrease in M protein concentration) were included, 11 of 16 patients (69%) responded to therapy. Major grade 3-4 toxicities included two patients with somnolence, and one patient each with syncope and neutropenia. Pre-treatment MVD was not a significant predictor of response to therapy, median MVD 4 and 12 in responders and non-responders respectively, P = 0.09. We conclude that thalidomide has significant activity in the treatment of newly diagnosed SMM/IMM. However, we do not recommend treatment with thalidomide at this stage since some patients with SMM/IMM can be stable for several months or years without any therapy. Additional randomized trials are needed to determine if thalidomide will delay progression to active multiple myeloma. PMID- 11480572 TI - Phage display mutagenesis of the chimeric dual cytokine receptor agonist myelopoietin. AB - Myelopoietins comprise a class of chimeric cytokine receptor agonists consisting of an hIL-3 (human interleukin-3) receptor agonist and an hG-CSF (human granulocyte colony-stimulating factor) receptor agonist linked head-to-tail at their respective carboxy and amino termini. The combination of an early acting cytokine (hIL-3) with a late acting one (hG-CSF) allows efficient hematopoeitic reconstruction following myeloablative insult, and drives differentiation of non myelocytic lineages (ie thrombocytic lineages) that are inaccessible using hG-CSF alone, in both preclinical models and clinical settings. A myelopoietin species was displayed and mutagenized on filamentous bacteriophage: both component agonists of myelopoietin were presented in biologically functional conformations as each recognized its corresponding receptor. Five amino acid positions in a short region of the hG-CSF receptor agonist module of myelopoietin that had been identified as important for proliferative activity were mutagenized. Display was used because it allows very 'deep' mutagenesis at selected residues: >10(5) substitution variants were affinity-screened using the hG-CSF receptor and 130 new, active variants of myelopoietin were identified and characterized. None of the selected variants were significantly more active than the parental myelopoietin species in a hG-CSF-dependent cell proliferation assay, though many were as active. Many of these relatively high-activity variants contained parental amino acids at several positions, suggesting the parental sequence may already be optimal at these positions for the assays used, and potentially accounting for the failure to identify enhanced bioactivity variants. Analysis of substitutions of high-activity variants complements and extends previous alanine scanning, and other genetic and biochemical data for hG-CSF variants. PMID- 11480573 TI - Cloning of human thymic stromal lymphopoietin (TSLP) and signaling mechanisms leading to proliferation. AB - Thymic stromal lymphopoietin (TSLP) is a novel cytokine that was found to promote the development of murine B cells in vitro. Here we describe the cloning and characterization of the human homologue of murine TSLP. This protein, which is expressed in a number of tissues including heart, liver and prostate, prevented apoptosis and stimulated growth of the human acute myeloid leukemia (AML)-derived cell line MUTZ-3. Anti-interleukin (IL)-7 receptor antibodies (Abs) neutralized this effect indicating that TSLP binds to at least part of the IL-7 receptor complex. TSLP induced phosphorylation of signal transducer and activator of transcription (STAT)-5. In contrast to IL-7, TSLP-triggered STAT-5 phosphorylation was not preceded by activation of janus kinase (JAK) 3. These findings would be in accordance with the notion, raised previously for the mouse system, that TSLP leads to STAT-5 phosphorylation by activating other kinases than the JAKs. Some other signaling pathways stimulated by many cytokines are not involved in TSLP activity; thus, TSLP did not stimulate activation of ERK1,2 and p70S6K. Furthermore, neutralizing Abs raised against cytokines known to stimulate the growth of MUTZ-3 cells did not inhibit the proliferative effects of TSLP, suggesting that TSLP-induced growth was a direct effect. In summary, we describe the cloning of human TSLP and its proliferative effects on a myeloid cell line. TSLP-induced proliferation is preceded by phosphorylation of STAT-5, but not of JAK 3. PMID- 11480574 TI - Paired multiplex reverse-transcriptase polymerase chain reaction (PMRT-PCR) analysis as a rapid and accurate diagnostic tool for the detection of MLL fusion genes in hematologic malignancies. AB - The MLL gene in chromosome band 11q23 is frequently rearranged in acute lymphoblastic and acute myeloid leukemias. To date, more than 50 different chromosomal regions are known to participate in translocations involving 11q23, many of which affect MLL. The pathogenetically important outcome of these rearrangements is most likely the creation of a fusion gene consisting of the 5' part of the MLL gene and the 3' end of the partner gene. Although abnormalities of the MLL gene as such are generally associated with poor survival, recent data suggest that the prognostic impact varies among the different fusion genes generated. Hence, detection of the specific chimeric gene produced is important for proper prognostication and clinical decision making. We have developed a paired multiplex reverse-transcriptase polymerase chain reaction analysis to facilitate a rapid and accurate detection of the most frequent MLL fusion genes in adult and childhood acute leukemias. To increase the specificity, two sets of primers were designed for each fusion gene, and these paired primer sets were run in parallel in two separate multiplex one-step PCR reactions. Using the described protocol, we were able to amplify successfully, in one single assay, the six clinically relevant fusion genes generated by the t(4;11)(q21;q23) [MLL/AF4], t(6;11)(q27;q23) [MLL/AF6], t(9;11)(p21-22;q23) [MLL/AF9], t(10;11)(p11-13;q23) [MLL/AF10], t(11;19)(q23;p13.1) [MLL/ELL], and t(11;19)(q23; p13.3) [MLL/ENL] in cell lines, as well as in patient material. PMID- 11480575 TI - T-lymphocytes in bone marrow samples of children with acute lymphoblastic leukemia during and after chemotherapy might hamper PCR-based minimal residual disease studies. PMID- 11480576 TI - Three further cases of t(8;14)(q11.2;q32) in acute lymphoblastic leukemia. PMID- 11480577 TI - Use of serum-free media to minimize apoptosis of chronic lymphocytic leukemia cells during in vitro culture. PMID- 11480578 TI - Acute myeloid leukemia with 17p abnormality in untreated essential thrombocythemia. PMID- 11480579 TI - Exposure to hydroxyurea during pregnancy: a case series. PMID- 11480580 TI - Intraventricular thrombosis during all-trans retinoic acid treatment in acute promyelocytic leukemia. PMID- 11480581 TI - Familial polycythemia vera: results from the Swedish Family-Cancer Database. PMID- 11480582 TI - Cellular and molecular characteristics of basal cells in airway epithelium. AB - Basal cells exist as a separate layer of cells covering most of the airway basal lamina. In this central position, they can interact with columnar epithelium, neurons, basement membrane, and the underlying mesenchymal cells. In addition, they interact with inflammatory cells, lymphocytes and dendritic cells. These interactions take place in the lateral intercellular space between basal cells. In this central position basal cells become a very important part of the epithelial-mesenchymal trophic unit of larger airways. In this review it is shown that basal cells may function as progenitor cells of airway epithelium and have a role in attachment of columnar epithelium with the basement membrane. They also have the potential to function in regulation of neurogenic inflammation, the inflammatory response, transepithelial water movement, oxidant defense of the tissue and formation of the lateral intercellular space. Other characteristics of basal cells were not clearly associated with a particular function. The functions for basal cells listed attempt to explain the presence of recently identified molecules in basal cells of airway epithelium. It should be pointed out that specific studies have not been carried out which test the relationship between the molecular functions we describe in this review and the basal cell in airway epithelium. PMID- 11480583 TI - Role of intercellular adhesion molecule 1 in acute lung injury induced by candidemia. AB - Candidemia, a complication often affecting immunocompromised patients, is a common cause of acute lung injury. Yeast-phase Candida albicans has been shown to express a protein that is antigenically and structurally related to Mac-1. C. albicans is reported to stimulate intercellular adhesion molecule 1 (ICAM-1) expression on endothelial cells. In this study, the authors examined the role of ICAM-1 in acute lung injury induced by candidemia. The authors cultured rat pulmonary artery endothelial cells (RPAEC) and investigated the effect of anti ICAM-1 antibodies on adhesion of C. albicans to RPAEC. In addition, the authors administered anti-ICAM-1 antibodies to rats to examine the effect of the antibodies on experimentally induced candidemia. Survival rates, lung wet-to-dry (W/D) weight ratios, bronchoalveolar lavage (BAL) fluid, histopathological findings, and colony-forming units (CFUs) of lung C. albicans were examined. The adherence of C. albicans to RPAEC was significantly decreased by anti-ICAM-1 antibodies. Anti-ICAM-1 antibodies significantly increased survival, decreased lung W/D weight ratios, decreased neutrophil counts in the BAL fluid, reduced microscopic lung injury, and decreased the quantity of lung C. albicans. These results indicate that ICAM-1 plays a role in adherence of C. albicans to pulmonary vascular endothelial cells, which likely leads to invasion of lung tissue by the organism. PMID- 11480584 TI - Pulmonary immunity to ragweed in a Beagle dog model of allergic asthma. AB - To create an allergy model in the dog, allergic Beagles with high levels of serum immunoglobulin E (IgE) and eosinophilia were bred; resulting puppies were sensitized to ragweed by intraperitoneal (IP) injection within 24 hours of birth through 22 weeks of age. At least 50% of the puppies developed high levels of serum IgE and eosinophilia. As young adults, 6 of these dogs, and 6 control age matched, nonallergic, nonimmunized dogs were exposed by inhalation to ragweed twice at 13-day intervals, and a third time 45 days later. Total and ragweed specific serum IgE and ragweed-specific serum IgG were increased significantly in allergic dogs relative to baseline. Allergic dogs had significantly greater levels of antibody specific for ragweed, as well as higher eosinophil counts in the bronchoalveolar lavage fluid, compared to nonallergic dogs. Airway reactivity to histamine in allergic, but not nonallergic dogs, increased significantly after aerosol exposure to ragweed. After a third exposure to ragweed, airway responses to histamine were elevated in the allergic dogs and remained high for at least 5 months. These results demonstrate the potential of the allergic dog model for investigating the underlying pulmonary immune mechanisms and therapeutic treatment of allergic asthma. PMID- 11480585 TI - Beta1-adrenoceptor stimulation by high-dose terbutaline downregulates terbutaline stimulated alveolar fluid clearance in ex vivo rat lung. AB - Because high-dose terbutaline and isoproterenol (10(-3) M), beta2-adrenergic agonists, failed to increase alveolar fluid clearance, the mechanisms responsible for this effect were examined in ex vivo rat lungs. An isosmolar 5% albumin solution with Evans blue dye was instilled into the distal airspaces in isolated rat lungs that were then inflated with 100% oxygen at an airway pressure of 8 cm H2O in a 37 degrees C incubator. Alveolar fluid clearance was measured by the progressive increase in dye concentrations over 1 hour. The results indicated that: (1) although 10(-5) M terbutaline or isoproterenol increased alveolar fluid clearance, 10(-3) M terbutaline or isoproterenol did not; (2) both concentrations of terbutaline (10(-5), 10(-3) M) increased intracellular adenosine 3',5'-cyclic monophosphate in cultured type II alveolar epithelial cells; (3) instillation of atenolol, a selective beta1-adrenergic antagonist, in the presence of either 10( 3) M terbutaline or isoproterenol was associated with an increase in alveolar fluid clearance. These results suggested that beta1-adrenoceptor stimulation prevented the normal response to a beta2-adrenergic agonist. To further test this hypothesis, a selective beta1-adrenergic agonist, denopamine, was administered; these results showed that (4) 10(-3) M denopamine, a selective beta1-adrenergic agonist, inhibited the increase in alveolar fluid clearance in the presence of 10(-5) M terbutaline; (5) hypoxia for 2 hours did not alter the effects of terbutaline on alveolar fluid clearance. The mechanism for the inability of the alveolar epithelium to respond to high-dose terbutaline or isoproterenol with the normal upregulation of alveolar fluid clearance in ex vivo rats lungs appears to be mediated by beta1-adrenoceptor stimulation that subsequently suppresses the beta2-adrenergic response. PMID- 11480586 TI - UVB light and 17-beta-estradiol have different effects on the mRNA expression of Ro/SSA and La/SSB autoantigens in HaCaT cells. AB - Antibodies produced against the Ro/SSA and La/SSB autoantigens are not only of diagnostic value but they may even play a role in the pathogenesis of several autoimmune diseases (Sjogren's syndrome, subacute cutaneous lupus erythematosus, neonatal lupus erythematosus and systemic lupus erythematosus). Among other factors, ultraviolet (UV) radiation and also the hormonal milieu are well-known cofactors in the pathogenesis of these autoimmune diseases. The goal of our research was to study the possible alterations in mRNA levels of three different Ro antigens and that of two La species produced by alternative splicing in transformed human keratinocytes (HaCaT cells) after UVB irradiation and after 17 beta-estradiol treatment. The polymerase chain reaction technique was used to determine the mRNA levels of the Ro and La species after 24, 48, and 72 h of irradiation. The mRNA levels of calreticulin increased as a function of time after UV irradiation but the mRNA levels of 52 kDa and 60 kDa Ro mRNAs were unaltered. After treating the cells with 17-beta-estradiol, there was no change observed in the levels of Ro mRNAs or La exon 1 mRNA, but a gradual decrease was noted in the mRNA levels of La exon 1'. The importance of alterations in the ratio of La exon 1 to exon 1' is supported by the observations in patients with Sjogren's syndrome, and our results strengthen the notion that the Ro and La antigens participate in the pathogenesis of different autoimmune diseases. PMID- 11480587 TI - Isolation of invasion-associated cDNAs in melanoma. AB - Metastasis and invasion are key steps in the systemic spread of tumor cells. To identify the genes involved in this process we recently selected highly invasive and weakly invasive cell clones from a melanoma cell line. Both cell clones showed a stable phenotype over more than 40 passages and previous analyses revealed a fivefold difference in their invasive potential in vitro and in tumorigenesis in vivo. To compare gene expression of the two cell clones a cDNA array system (Clontech human cancer cDNA array) was used. Exact quantification of differentially expressed genes in each cell clone was performed by real-time RT PCR. An evaluation of the array data revealed a total of 36 genes that were more than 1.5-fold differentially expressed, and 26 (72%) of these showed a differential expression pattern by quantitative RT-PCR. Previously known differences in expression patterns, including loss of p16 and HLA I, or equal expression of p73, and RAR alpha, beta and gamma were confirmed by the array data. In addition, reduced expression levels of several cytoskeletal proteins, such as vimentin, gamma-actin, desmin and cytokeratins, in the highly invasive cell clone were reproducibly identified. Other genes strongly upregulated in the highly invasive cell clone included jagged 2, STAT1, tPA and c-myc, whereas MDA-6 (p21), caspase 2 and semaphorin were found to be downregulated. In conclusion, comparative hybridization of cDNA arrays identified a series of novel invasion associated changes in gene expression and confirmed previously known expression patterns. PMID- 11480588 TI - Effect of nerve growth factor on endothelial cell biology: proliferation and adherence molecule expression on human dermal microvascular endothelial cells. AB - In addition to its effect on the central nervous system, nerve growth factor (NGF) appears to play a key role in the initiation and maintenance of inflammation in many organs. NGF degranulates mast cells, recruits inflammatory cellular infiltrates and activates T cells. Extravascular migration of leukocytes is initially controlled by the interaction of cell surface adhesion molecules of leukocytes and endothelial cells. A marked upregulation of NGF in keratinocytes is also observed in conditions characterized by angiogenesis such as psoriasis and wound healing. In this study we investigated the role of NGF in inflammation by studying its effects on endothelial cell proliferation and intracellular adhesion molecule expression by endothelial cells. The effect of NGF on human dermal microvascular endothelial cell (HDMEC) proliferation was measured using the hexosaminidase assay. ICAM-1 expression on HDMEC was measured by ELISA. The function of ICAM-1 was assessed by adherence of peripheral blood mononuclear cells (PBMC) to HDMEC using 51Cr-labeled PBMC. There was a significant increase in proliferation of HDMEC stimulated with NGF as compared to unstimulated HDMEC (P < 0.001). NGF-neutralizing antibody decreased the mitogenic effect of NGF significantly (P < 0.05). NGF also increased ICAM expression on HDMEC as compared to unstimulated HDMEC (P < 0.05). NGF-neutralizing antibody decreased ICAM expression on NGF-stimulated HDMEC (P < 0.05). The percentage of PBMC adherence was higher in NGF-stimulated HDMEC (P < 0.001). Anti-ICAM antibody decreased PBMC adherence. In the study reported here, the role of NGF in two important aspects of inflammation, i.e. angiogenesis and inflammatory cell recruitment at the site of inflammation, was investigated. PMID- 11480589 TI - Expression of vascular endothelial growth factor in a human hemangiosarcoma cell line (ISO-HAS). AB - Vascular endothelial growth factor (VEGF), in addition to being a specific mitogen of endothelial cells in vitro, is also known to induce angiogenesis in vivo. These functions suggest that VEGF may play an important role in the growth of hemangiosarcomas. Previous studies have demonstrated the expression of VEGF and its receptors, flt-1 or KDR/flk-1, in hemangiosarcomas by immunohistochemical staining and in situ hybridization. In the present study, however, we demonstrated that tumor cells of the hemangiosarcoma cell line ISO-HAS express mRNA of VEGF and its two receptors, flt-1 and KDR, and secrete VEGF protein. VEGF mRNA expression and protein secretion were found to be enhanced by phorbol 12 myristate 13-acetate. In addition, we demonstrated that ISO-HAS cells respond to recombinant human VEGF165 with a dose-dependent up-regulation of cell proliferation and growth. These results suggest that the VEGF-VEGF receptor system plays a role in proliferation and growth of hemangiosarcoma cells. PMID- 11480590 TI - Molecular weight dependence of polyethylene glycol penetration across acetone disrupted permeability barrier. AB - Previous studies have demonstrated that permeability barrier disruption by acetone treatment significantly enhances skin permeability to both hydrophilic and amphipathic compounds, but not to highly lipophilic compounds. The purpose of the present study was to investigate the dependence of permeability on molecular weight (MW) in acetone-disrupted hairless mouse skin in contrast to normal skin. Penetration of polyethylene glycol (PEG) 300, 600, and 1,000 over 12 h was measured using diffusion cells. High-performance liquid chromatographic methods with refractive index detection were used to separate and quantitate the individual oligomeric species in the PEG samples. Percutaneous penetration of PEGs exhibited slightly steeper MW dependency at a transepidermal water loss (TEWL) of 30-41 g/m2 per h in comparison with TEWLs of 0-10 (control skin), 10 20, and 20-30 g/m2 per h, with a higher percentage of smaller oligomer PEGs penetrating than larger ones. Increasing the TEWL of the skin increased the penetration of all the PEG oligomers, and the degree of the enhancement relative to penetration through control skin increased with MW and was maximal for oligomers with a MW ranging from 326 to 414 Da. Within the limit of quantitation of the assay, the MW cut-off for PEG penetration across mouse skin with TEWLs of 0-10, 10-20, and 20-30 g/m2 per h was 414, 590, and 942 Da, respectively, while all the measurable oligomers up to MW 1,074 Da were able to penetrate skin with TEWLs in the range 30-41 g/m2 per h. The results suggest that not only higher amounts but also more varieties of chemicals may penetrate skin with a compromised barrier than normal skin, implying a higher risk of intoxication and hypersensitization by environmental agents through diseased skin with impaired barrier function. PMID- 11480591 TI - Functional and structural changes of the epidermal barrier induced by various types of insults in hairless mice. AB - Acute barrier disruption, regardless of the method of induction, depletes the stratum corneum intercellular lipids and this stimulates a series of lipid/ DNA synthesis activities which lead to barrier recovery. After barrier disruption by tape stripping, occlusion with a water vapor-impermeable membrane inhibits barrier repair. In this study, we investigated the changes in the murine epidermis after barrier perturbation by tape stripping and three different types of surfactants (Emalex NP-12, ENP-12; sodium dodecyl sulfate, SDS; benzalkonium chloride, BKC). To examine the effect of an artificial barrier, we covered the animals with a water vapor-impermeable membrane for 3 days following barrier disruption and then exposed them to the air for 2 days. The histological findings after occlusion or air exposure were similar. However, after air exposure for 2 days, the thickness of the epidermis including the stratum corneum and the stratum granulosum layers decreased to about half that of the epidermis after occlusion. Ultrastructural examination revealed obvious distortion of the lamellar bilayers within the stratum corneum interstices immediately after barrier disruption. After 3 days of occlusion, extensive disorganization was evident in the intercellular domain of the stratum corneum, whereas 2 days after removal of the occlusion, the normal basic unit structure of the lamellar bilayers had partially reappeared. Our findings provide evidence that the kinetic pattern of barrier repair and the morphological changes are similar after occlusion following barrier disruption regardless of the mechanism of disruption. PMID- 11480592 TI - Decreased levels of CXCR3 transcripts in peripheral blood mononuclear cells from patients with atopic dermatitis and with cutaneous diseases associated with eosinophilia. PMID- 11480593 TI - Hip abductor strength following total hip arthroplasty: a prospective comparison of the posterior and lateral approach in 100 patients. AB - We studied the hip abductor strength and Trendelenburg test prospectively in 100 patients undergoing total hip replacement via a lateral or posterior approach. In 49 patients, we used the lateral approach to implant the Charnley total hip replacement, and in 51 patients, the posterior approach to implant the Exeter total hip. Isometric abductor strength was measured with the kinetic communicator device and the Trendelenburg test was recorded preoperatively and at 3 and 12 months postoperatively. Of the original 100 patients, 83 were available for study at 3 months and 73 at 12 months. Hip abductor strength and the Trendelenburg test improved postoperatively in both groups, but we found no difference in hip abductor strength recovery at 3 and 12 months between the lateral approach and the posterior approach. Similarly there was no difference in the Trendelenburg test between the two groups 3 and 12 months following hip replacement. PMID- 11480594 TI - Acetabular revision with impacted grafting and a reinforcement ring: 42 patients followed for a mean of 10 years. AB - We analyzed the outcome of 42 cemented cup revisions (40 patients) with morselized bone grafts and a reinforcement ring after median 10 (7-16) years. The acetabular component migrated a mean of 7 (5-8) mm in 5 hips. The migration stopped after 2 years except in 1 hip, the only one revised because of mechanical loosening. 3 other hips were revised because of infection. Among the other 38 hips (36 patients), the Merle d'Aubigne score increased from 9 to 15 points. PMID- 11480595 TI - Femoral alignment of the Charnley stem: a randomized trial comparing the original with the new instrumentation in 123 hips. AB - Deficient cement mantles are associated with aseptic loosening of the stem component of total hip replacement. In a former study of 206 Charnley stems, we found high frequencies of stem malalignment, especially on the lateral view, consequently resulting in a high percentage of low cement mantle grading. If the "true" lateral radiographic projection is not used, there is a risk that the frequency of mantle defects is underestimated. A logistic regression analysis showed a high correlation between low cement mantle grading and stem loosening after a mean follow-up of 10 years. The new Charnley instrumentation was introduced in 1994 and we started a randomized trial including 123 prostheses to determine whether the new instrumentation improved the position of the stem in both the AP and lateral planes. Postoperative radiographs revealed a significant change in AP positioning-i.e., from a high percentage of varus with the original method to valgus with the new instrumentation. However, there was no difference on the lateral view, with a persisting high frequency of stems with implant-inner cortex contact resulting in high percentages of low cement-mantle grading in both systems. If this deficiency, in a long-term perspective, is associated with aseptic loosening, as many authors have claimed, the manufacturers should address the problem. PMID- 11480596 TI - Charnley total hip arthroplasty in ankylosing spondylitis: survivorship analysis of 76 patients followed for 8-28 years. AB - 76 consecutive Charnley low friction hip arthroplasties were performed in 54 (37 men) patients with ankylosing spondylitis from 1971 to 1991 in the Rheumatism Foundation Hospital. Their mean age was 40 (16-67) years. They were followed until the end of 1999. The overall prosthesis survival was 80% at 10 years, 66% at 15 years and 62% at 20 years. The survival of the acetabular component was 91%, 77% and 73% at 10, 15 and 20 years and that of the femoral component 82%, 79% and 77%, respectively. We found no significant risk factor that predicted prosthesis survival. PMID- 11480597 TI - Effective bactericidal activity of tobramycin and vancomycin eluted from acrylic bone cement. AB - We studied the bioactivity of vancomycin and tobramycin eluted from methylmethacrylate bone cement. Aliquots of the drainage were obtained at 1, 6, 12 and 24 hours following total hip prosthetic implantation with vancomycin tobramycin-loaded cement in 3 patients. The samples were analyzed with fluorescence polarization immunoassay and bioassay, using group B streptococcus for vancomycin and Escherichia coli for tobramycin. These bacteria were selected due to the effectiveness of vancomycin and poor effectiveness of tobramycin against group B streptococcus and conversely with E. coli. The immunodetection of vancomycin averaged 14 (1 hour), 9 (6 hours), 10 (12 hours) and 11 microg/mL (24 hours). The bioassay averaged 47, 36, 79 and 41 microg/mL (p = 0.03). The immunodetection of tobramycin averaged 43, 21, 18 and 14 microg/mL; and bioassay 30, 15, 15 and 12 microg/mL (p = 0.1). Both antibiotics eluted with a highly effective bactericidal activity. Our findings indicate that the presence of tobramycin has a synergistic-like effect on the bactericidal activity of vancomycin, which has not been previously reported. We recommend a combination of vancomycin and tobramycin with cement for the treatment of orthopedic infections caused by gram-positive organisms. PMID- 11480598 TI - No lymphokines in T-cells around loosened hip prostheses. AB - Research results have been contradictory about the role of lymphocytes and immune response in aseptic loosening of total hip replacement (THR). Conclusive evidence is still lacking in spite of extensive in vivo and in vitro studies. Our study was designed to check whether T-cells were activated and if they produced lymphokines in synovial membrane-like interface tissue around loosened THRs. Tissue sections were stabilized and permeabilized to allow the cytokine-specific antibodies to penetrate through the cell membrane and the membranes of intracellular organelles. This technique, combined with computer-assisted image analysis, permits the detection and quantitation of lymphokine-producing cells. We found that the number of T-cells was low, and none of the T-cells was activated, as shown by the absence of interleukin-2 receptor (IL-2R) immunoreactivity. There was no cell producing lymphokines, such as interleukin-2 (IL-2), interferon-gamma (IFN-gamma), and tumor necrosis factor-beta (TNF-beta). Our results suggest that T-cell-mediated immune response is not actively involved in aseptic loosening of THR. PMID- 11480599 TI - Changes in biochemical markers of bone metabolism and BMD during the first year after a hip fracture. AB - We measured the levels of biochemical markers of bone formation and bone resorption in hip fracture patients preoperatively and after 6 and 12 months. Bone densitometry was done with quantitative computer tomography (QCT), dual energy X-ray absorptiometry (DXA) and heel ultrasound. After 6 months, the biochemical markers of bone formation and bone resorption had increased. The levels remained high after 1 year and no change occurred between 6 and 12 months. We found no correlations between biochemical bone markers and bone density/stiffness on admission and change in bone mineral density (BMD) during the first postoperative year, despite the changes in bone markers and bone density. In our opinion, biochemical bone markers can not be used to predict bone loss in the individual patient after a hip fracture. PMID- 11480600 TI - More postoperative femoral fractures with the Gamma nail than the sliding screw plate in the treatment of trochanteric fractures. AB - Despite several studies showing a higher incidence of peri-implant femoral fractures with the Gamma nail than with a sliding screw plate (SSP), the Gamma nail has remained the standard implant for trochanteric fractures in many hospitals. We recorded 921 trochanteric fractures in the city of Oslo during 2 years and compared the reoperation frequency in patients treated with the Gamma nail (n 379) and SSP (n 542). The distribution of age and gender in the two treatment groups was the same. 65 patients were reoperated on, several of them more than once. The only significant difference between the two surgical methods in complications leading to a reoperation was the frequency of femoral shaft fractures. 17 of the patients treated with the Gamma nail had a new femoral fracture postoperatively, compared to 3 of those with a SSP. The relative risk of another femoral fracture after surgery was 12 (95 % CI: 2.7-52) if the surgical device was a Gamma nail compared to a SSP. The Gamma nail therefore can not be recommended as the standard implant for trochanteric fractures. PMID- 11480601 TI - Flat vs. concave tibial joint surface in total knee arthroplasty: randomized evaluation of 39 cases using radiostereometry. AB - 40 patients with non-inflammatory arthrosis and minor preoperative deformity (< or =5 degrees ) were operated on with an AMK type (DePuy, Johnson & Johnson) total knee arthroplasty (TKA). The posterior cruciate ligament was retained. The patients were divided into those with a flat (terminology of the manufacturer: standard) or a concave (terminology of the manufacturer: constrained) polyethylene insert (20 in each group). Radiostereometric (RSA) examinations were done postoperatively and after 3,12 and 24 months. The median absolute rotations of the tibial inserts varied between 0.12 and 0.24 (range 0.00-1.54) degrees, with no differences between the 2 groups. The median maximum total-point motions (flat/concave = 0.41/0.42 mm), the maximum subsidence or lift-off did not differ. The Hospital for Special Surgery knee score and the patients' opinion about the operation, based on their preoperative expectations, showed little, if any, differences. At 2 years, 10 of 20 patients with flat and 13 of 19 with concave inserts regarded their knee function as normal or almost so. PMID- 11480602 TI - Macroscopic and histological assessments of the cruciate ligaments in arthrosis of the knee. AB - We examined the macroscopic appearance of both cruciate ligaments in 52 knees during knee replacement. It was classsified as normal, abnormal or ruptured. The ligaments were also evaluated histologically: stage 0 (normal), stage I (degeneration of < 1/3 of the collagen fibers), stage 2 (degeneration of 1/3-2/3) and stage 3 (> 2/3). 17 anterior cruciate ligaments (ACL) were normal, 14 were abnormal and 21 ruptured. All the posterior cruciate ligaments (PCL) were normal. 14 ACL were stage 0, 6 stage 1, 8 stage 2 and 24 were stage 3. 22 PCL were stage 0, 14 stage 1, 13 stage 2 and 3 were stage 3. When the ACL was abnormal or ruptured, the PCL was stage 0 only in one fourth of the cases. The long-term results of TKR retaining the PCL should be better if the ligament is strong. The intraoperative assessment of the macroscopic appearance of the ACL reflects the histological state of the PCL. PMID- 11480603 TI - Increased interleukin-6 in collected drainage blood after total knee arthroplasty: an association with febrile reactions during retransfusion. AB - We determined interleukin-6 (IL-6) concentrations in collected shed drainage blood intended for retransfusion in a prospective study in 81 patients after total knee replacement. We found large increases in IL-6 levels, averaging 6.5 (SD 3.9) ng/mL, in shed blood collected in the first 6 postoperative hours. 3 patients had febrile reactions after collected blood was retransfused. The IL-6 levels in the drainage blood of these patients were very high (9.6-13.4 ng/mL). In the blood collected after 6 hours, IL-6 concentrations increased to 47 (SD 33) ng/mL (p < 0.001). These results suggest a relation between increased interleukin 6 concentrations in shed drainage blood and the occurrence of febrile reactions after retransfusion of such blood. PMID- 11480604 TI - Vertebral deformation in urban Swedish men and women: prevalence based on 797 subjects. AB - Vertebral fracture-deformation, a common feature of osteoporosis, shows considerable age, sex and geographical variation. We present the prevalence in an urban population of south-west Sweden. Lateral spine radiographs of 797 men and women, age 50-86 years, were evaluated by morphomety. The age-standardized prevalence of subjects with vertebral deformation using the deformation criterion -3 SD was 39 (95%CI 34-43)% in women and 33 (95%CI 28-38)% in men. The prevalence increased with age in both sexes. After adjustment for age, women had a higher prevalence than men, odds ratio 1.4. The proportion of vertebrae with deformation ranged from 2%-11%, increasing with age. The vertebrae most commonly deformed were Th 11, Th 12 and L1. PMID- 11480605 TI - Increased median nerve latency at the carpal tunnel of patients with "trigger finger": comparison of 62 patients and 13 controls. AB - An association between symptomatic compression neuropathy of the median nerve at the carpal tunnel and "trigger finger" has been reported in endocrine and metabolic disorders. We assessed the incidence of increased median nerve latency in subjects with "trigger finger". 62 consecutive patients with "trigger finger" and no signs or symptoms of median nerve compression underwent nerve conduction studies of the median nerve. 13 healthy adults served as controls. 39/62 patients had increased distal motor latency in the median nerve. Only 1 of 13 subjects in the control group had a borderline value of distal motor latency. PMID- 11480606 TI - Rabbit supraspinatus tendon detachment: effects of size and time after tenotomy on morphometric changes in the muscle. AB - We studied the effects of size and time after rabbit supraspinatus tenotomy (group A--small tenotomy, group B--large tenotomy) on muscle morphometric changes in 48 rabbits. Animals were killed 6 (subgroups A1 and B1), 12 (subgroups A2 and B2) and 24 weeks (subgroups A3 and B3) after tendon detachment. Statistically significantly greater increases in interstitium volume were noted in subgroups A1 A3 and B1-B3 than in controls. Reductions in type I and II fiber diameters were mainly due to the length of observation. However, statistically significant differences in comparison with controls appeared earlier after large tenotomy. The size of the tenotomy primarily affected muscle fiber composition. Our results suggest that these changes were caused by fiber transformation from type I to type II and vice versa. These findings indicate that the interdigitations between the supraspinatus and infraspinatus and between the supraspinatus and subscapularis tendons are important in dynamics and the degree of morphometric changes in the rabbit supraspinatus after tenotomy. PMID- 11480607 TI - Dose-related cellular effects of platelet-derived growth factor-BB differ in various types of rabbit tendons in vitro. AB - Cellular responsiveness to growth factors that can affect tendon healing may be site-specific. We have compared the dose-response effects of platelet-derived growth factor-BB (PDGF-BB) on proteoglycan, collagen, noncollagen protein and DNA synthesis between intrasynovial intermediate and proximal segments of deep flexor tendons, and extrasynovial peroneal tendons of rabbits during short-term cultures. PDGF-BB stimulated matrix and DNA synthesis of the three types of tendon segments in a dose-dependent manner in the range from 0.1 to 100 ng/mL. PDGF-BB stimulated collagen synthesis and noncollagen protein synthesis (calculated from LogED50) in proximal intrasynovial tendon segments more than in extrasynovial peroneal tendon segments, and DNA synthesis less in proximal than in intermediate intrasynovial tendons. However, the estimated maximal stimulations (Emax) by PDGF-BB were similar in the three types of tendon segments. These findings show that PDGF-BB stimulates DNA and matrix synthesis differently in various types and regions of tendons during short term explant culture and suggests that there may be differences in cellular responsiveness during tendon healing. PMID- 11480608 TI - Oxaceprol, an atypical inhibitor of inflammation, reduces leukocyte adherence in mouse antigen-induced arthritis. AB - Oxaceprol (N-acetyl-L-hydroxyproline), an atypical inhibitor of inflammation, is an established drug forjoint disease without serious side-effects. Recent studies have emphasized that oxaceprol has an effect on the microcirculation. Since the exact mechanism of action remains unclear, the aim of our study was to investigate the leukocyte-endothelial cell interactions in oxaceprol-treated mice with antigen-induced arthritis (AiA) using intravital microscopy. In our study, Balb/c mice were allocated to 4 groups (n 7, 8, 8, 8): 2 control groups with saline or oxaceprol and 2 groups of arthritic animals which received saline or oxaceprol (100 mg/kg twice a day intraperitoneally). The severity of arthritis was quantified by the transverse knee joint diameter. For the intravital fluorescence microscopy measurements on day 10 after inducing arthritis, the patella tendon was partily resected to visualize the intraarticular synovial tissue of the knee joint. The number of rolling and adherent leukocytes as well as RBC velocity and functional capillary density (FCD) were quantified in synovial microvessels. Furthermore, leukocyte infiltration was determined in the histological sections with an established score. No significant changes in mean arterial blood pressure or functional capillary density were found in any of the groups. However, the leukocyte rolling fraction and number of leukocytes adherent to the endothelium were increased in postcapillary venules of the synovium in arthritic animals (0.16 to 0.31, 78 cells/mm2 to 220 cells/mm2). In animals with AiA treated with oxaceprol, leukocyte adherence and swelling were significantly reduced in comparison to the arthritic animals treated with saline. Furthermore, the histological score showed less leukocyte infiltration in the oxaceprol treated arthritic animals. Thus, oxaceprol reduces leukocyte adherence in vivo and leukocyte infiltration in mouse AiA, indicating an effect on synovial microcirculation. PMID- 11480609 TI - Effects of hyaluronic acid and basic fibroblast growth factor on motility of chondrocytes and synovial cells in culture. AB - The capacity of chondrogenic precursor cells to migrate and proliferate in an injured area is considered to be essential for cartilage repair. We examined cell motility of chondrocytes and synovial cells in monolayer culture and the chemokinetic effects of hyaluronic acid (HA) and basic fibroblast growth factors (bFGF) on these cells. The velocity of chondrocyte migration was accelerated by giving bFGF and simultaneously administering of both HA and bFGF, but it was not affected by HA alone. The velocity of synovial cell migration was increased by HA, but not by bFGF. HA had a chemokinetic effect on synovial cells and bFGF had the same effect on chondrocytes. Treatment with exogenous HA and bFGF may be of value for repairing articular cartilage injury by recruiting chondrogenic cells and promoting migration of chondrocytes in the cartilage tissue. PMID- 11480610 TI - Increases in callus formation and mechanical strength of healing fractures in old rats treated with parathyroid hormone. AB - We studied the effects of intermittent administration of parathyroid hormone (PTH(1-34)) on callus formation and mechanical strength of tibial fractures in 27 month-old rats after 3 and 8 weeks of healing. 200 microg PTH(1-34)/kg was administered daily during both periods of healing, and control animals with fractures were given vehicle. At 3 weeks, PTH treatment increased maximum load and external callus volume by 160% and 208%; at 8 weeks, by 270% and 135%. It also enhanced callus bone mineral content (BMC) by 190% and 388% (3 and 8 weeks). From week 3 to week 8, callus BMC increased by 60% in the vehicle-injected animals, and by 169% in the PTH-treated animals. In the contralateral intact tibia, PTH treatment increased BMC by 18% and 21% (3 and 8 weeks). No differences in body weight were found between the vehicle-injected and the PTH-treated animals during the experiment. In conclusion, PTH treatment enhances fracture strength, callus volume and callus BMC after 3 and 8 weeks of healing. PMID- 11480611 TI - Pathophysiology of bone pain: a review. PMID- 11480612 TI - Dislocating medial head of triceps. PMID- 11480613 TI - The effect of infection with bovine viral diarrhea virus on the fertility of Swiss dairy cattle. AB - Bovine viral diarrhea virus is a major cattle pathogen with a worldwide distribution. Animals may be infected with BVD virus transiently or persistently. Transient infection leads to protective immunity. Persistent infection is unique because it is associated with an immunotolerance that is specific to the infecting strain of BVD virus. Persistent infection results from viral invasion of fetuses between the second and fourth month of development. Such animals are of prime importance in the epidemiology of BVD because they shed large amounts of virus, and thus serve as a constant source of infection for non-immune animals. Infection of pregnant animals during the first two months of gestation may result in an increased rate of return to estrus. Animals infected in the period of five months to birth may abort or give birth to calves with malformations. The effects of BVD virus on fertility and gestation are well documented from experimental infection. However, much less is known of the extent of losses under field conditions. The main reason for this may be the multitude of other causes of increased return rates and gestation failures. In addition, the incidence of infection with BVD virus may vary over time and depends on management practices. In this study, we investigated the impact of BVD virus on gestation failures under field conditions in a large cattle-breeding area of Switzerland, where no specific measures to control BVD are in effect. Our approach consisted of relating seroconversions to BVD virus with the rate of return to estrus, abortion, and birth of calves with apparent malformations. These parameters of fertility were compared to those of animals immune to BVDV infection due to previous exposure to the virus and animals without seroconversion. Our data show that infection with BVD virus during the first 45 days of gestation did not influence the rate of return to estrus. By contrast, we observed a statistically significant increase in the abortion rate in mid-term gestation (Days 46 to 210) while no such effect was observed in the later stages of gestation. No clinically manifest malformations were observed in the offspring of animals that had seroconverted to BVD virus. In our study population the prevalence of BVDV antibody positive cattle varied only slightly between 78% and 80% over the period of observation. Our data showed that 7% (CI: 2.4-14%) of fetal deaths may be attributable to infection with BVD virus. PMID- 11480614 TI - Monoclonal antibodies to intra-acrosomal proteins inhibit gamete binding in vitro. AB - The sperm-zona pellucida-binding assay in vitro was used as a functional test for zona pellucida-binding ability of boar spermatozoa after co-incubation with monoclonal antibodies against intra-acrosomal proteins. The effect of monoclonal antibodies ACR.2 against boar acrosin (55, 53, 45 and 38 kDa), and Hs-8 against boar intra-acrosomal protein (230, 110, 88, 60, 48 kDa) on boar spermatozoa porcine oocyte binding was examined. The sperm-zona pellucida-binding was reduced when medium was supplemented with monoclonal antibodies during sperm-oocyte co incubation, but not when capacitated spermatozoa were pretreated with monoclonal antibodies before incubation with oocytes. Our results show that the monoclonal antibodies (ACR.2, Hs-8) against intra-acrosomal proteins reduce the secondary sperm-zona pellucida-binding with statistically significant difference. This suggests the role of these proteins in the early phases of fertilization. PMID- 11480615 TI - Pentoxifylline improves in vitro fertilization and subsequent development of bovine oocytes. AB - The purpose of this study was to examine whether pentoxifylline improves in vitro fertilization and developmental rates of bovine oocytes. In the first experiment, we examined the effects on the fertilization rate of various concentrations of pentoxifylline (0-7.5 mM) combined with heparin (10 IU/mL). In the second experiment, we examined fertilization cleavage and blastocyst rates after frozen thawed spermatozoa, obtained from four different bulls, were incubated with heparin (10 IU/mL) with or without caffeine (5 mM) or pentoxifylline (5 mM). In the first experiment, a significantly higher fertilization rate was obtained in heparin containing 5 mM pentoxifylline compared to that in heparin alone or in heparin containing 7.5 mM pentoxifylline (86% vs 60% vs 64%, respectively). The percentage of monospermy in 5 mM pentoxifylline (81%) was significantly higher than in heparin alone (57%). In the second experiment, the interactions among Bulls A, B, C, and D; between treatments (pentoxifylline-with-heparin, caffeine with-heparin and heparin alone), and between bulls and treatments were analyzed for the number of oocytes penetrated, monospermic oocytes, cleaved oocytes and blastocysts. Among bulls, there was a significant difference in the number of oocytes penetrated (P < 0.01), monospermic oocytes (P < 0.05), cleaved oocytes (P < 0.001), and blastocysts (P < 0.001). Between treatments, there was a significant difference in the number of oocytes penetrated (P < 0.001), monospermic oocytes (P < 0.01) and cleaved oocytes (P < 0.001). Interaction between bulls and treatments was observed for the number of oocytes penetrated (P < 0.05). Individually, for Bulls A, C and D, the numbers of oocytes penetrated and monospermic oocytes in pentoxifylline-with-heparin were significantly higher than in heparin alone. For Bull D, significantly higher results were obtained for the number of oocytes penetrated, monospermic oocytes, cleaved oocytes and blastocysts in pentoxifylline-with-heparin compared to caffeine-with-heparin and heparin alone (P < 0.05). These results suggest that treating sperm with 5 mM pentoxifylline in combination with heparin is effective for bovine in vitro fertilization and it that this treatment is effective even for bulls that produce low fertilization and blastocysts after sperm treatment with caffeine-with heparin or heparin alone. PMID- 11480616 TI - Use of vaginal electrical resistance (VER) to predict estrus and ovarian activity, its relationship with plasma progesterone and its use for insemination in buffaloes. AB - In three experiments we studied the baseline and changes in VER during different natural estrous cycle stages (n=146) in ovarian structures and in plasma progesterone during estrus induced by prostaglandin injection (n=16) and the VER at insemination (n=90) in an attempt to predict estrus, ovulation and the best VER range for inseminating buffaloes for optimum conception. The baseline VER was classified on the basis of ovarian findings and estrous cycle stages. The mean VER during estrus, metestrus, diestrus, proestrus and anestrus was 32.68 +/- 0.46, 41.26 +/- 1.17, 50.23 +/- 0.55, 43.20 +/- 0.64 and 55.86 +/- 0.57 ohms, respectively. There was a significant difference (P<0.01) between the VER except those between metestrus and proestrus. The ANOVA for VER over estrous cycle stages showed a highly significant (P<0.01) effect of stage of estrous cycle on VER in buffaloes. The percent decrease in VER was more pronounced from diestrus to estrus. In the second part of the study plasma progesterone profiles and the appearance of estrus in buffaloes induced to estrus using two dose schedules and routes of PGF2alpha administration showed that luteolysis and estrus induction was slower in the 10 mg i.v.s.m. route (Intra Vulvo Submucosal) (only 60% animals evinced estrus in 48 to 72 hours) as compared to the 25 mg i.m. route (83.33% evidenced estrus in 48 to 72 hrs). Fall in plasma progesterone was synchronous to a fall in VER, the correlation (0.65) between them being positive and significant (P<0.01). After ovulation the VER started rising, showing a distinct relationship between VER and ovulation. By using VER, an additional 36.6% of the buffaloes could be detected in estrus. In the third part of the study, insemination of buffaloes induced to estrus (n=11) and normal-estrus buffaloes (n=79) showed that the overall conception rates to single insemination when the buffaloes were inseminated at the VER range of 26 to 30, 31 to 35 and 36 to 40 ohms were 81.48, 58.97 and 16.66%, respectively. Buffaloes showing VER from 31 to 35 ohms and 36 to 40 ohms also evidenced atypical and Null fern pattern in the cervicovaginal mucus. The study proved that VER can be used successfully to predict the stage of estrous cycle, ovarian status and ovulation; and insemination at a low VER distinctly improves the conception rates in buffaloes. PMID- 11480617 TI - Heparin and Ca2+-free medium can enhance release of bull sperm attached to oviductal epithelial cell monolayers. AB - The success of assisted reproductive techniques, such as IVF, could be enhanced by being able to select the most competent spermatozoa in a sample. Attachment and subsequent release of spermatozoa from oviductal epithelial cells (OEC) could provide populations of functionally superior spermatozoa for use in these protocols. The objective of the present study was to investigate the ability of heparin and Ca2+-free medium to induce spermatozoa release from bovine OEC. Epithelial cells were grown to confluence in 24-well plates and pooled frozen bull semen was added to a final concentration of 1 x 10(6) spermatozoa/well. Spermatozoa were allowed to bind to OEC for 2 h. Medium with unbound spermatozoa was removed and replaced by Sperm-TALP, only (control), with heparin (5, 10, or 15 IU/mL), or Ca2+-free with 2 mM EGTA. Treatments were left on sperm-OEC co cultures for 0.5, 1, 2, 3, or 5 h. At each time, the media were recovered and spermatozoa from each treatment were counted and evaluated for acrosome integrity and motility. The total number of spermatozoa attached to OEC after 2 h of co culture was considered 100%. Spermatozoa release is expressed as percentage of the total number of sperm cells bound to OEC after 2 h of co-culture. Data were analyzed by ANOVA and results are expressed as mean +/- SEM from three independent replicates. Beginning at 0.5 h, more sperm cells (P < 0.05) were released from OEC in the heparin groups (10 and 15 IU/mL, 77.3 +/- 6.2% and 84.0 +/- 6.2%, respectively) as compared to the control (46.4 +/- 6.2%). The Ca2+-free medium also induced spermatozoa release when compared with the control, but the effect was not significant until 3 h (38.2 +/- 1.9% vs 59.5 +/- 6.9%; P < 0.05). The percentage of acrosome reacted spermatozoa was not affected by heparin treatment. Heparin at 10 IU/mL increased (P < 0.05) the percentage of motile spermatozoa, whereas Ca2+-free medium caused the opposite effect at 0.5 h after addition of treatments. We conclude that both heparin and Ca2+-free medium are able to promote spermatozoa displacement from OEC attachment. Based on motility and acrosome status data, we predict that released sperm cells may be used for IVF and other assisted reproductive techniques. PMID- 11480618 TI - Assessment of in vitro fertility of deer spermatozoa by heterologous IVF with zona-free bovine oocytes. AB - We have previously shown that the in vitro development of deer embryos differed according to the IVF conditions. The aim of the study was to use heterologous IVF with zona-free matured bovine oocytes to assess the in vitro fertility of 3 samples of deer semen (2 ejaculates from sika deer (Cervus nippon) and 1 pool of epididymal spermatozoa from red deer (Cervus elaphus)). The frozen/thawed semen samples were selected on Percoll gradient and resuspended in Tyrode modified medium supplemented with estrus sheep serum (0, 2, 20% v/v) or heparin (10 microg/mL). During 8 h of culture, the sperm motility index according to the post insemination time (hpi) did not differ either between samples or between supplemented IVF media. In vitro matured zona-free bovine oocytes were inseminated in different IVF media with the semen samples. Penetration rates assessed at 15 hpi were optimal with 20% estrus sheep serum for sika deer ejaculates whereas 2% were sufficient to reach the maximum functionality of epididymal spermatozoa from red deer. The mean time of pronuclear formation was similar regardless of the semen sample. The precocity of the onset of the first S phase in both pronuclei was characterized by Bromo-deoxy-Uridine exposures between 5 and 15 hpi in order to assess the developmental potential conferred by the semen sample (intrinsic value). As we previously observed in homologous IVF, this value seemed to be higher for the epididymal sperm sample. PMID- 11480619 TI - Thiols prevent H2O2-mediated loss of sperm motility in cryopreserved bull semen. AB - We previously showed that cryopreservation of bull spermatozoa in egg yolk Tris extender (EYTG) significantly reduced the intracellular level of thiols. Other studies showed the beneficial effects of adding antioxidants to cryopreserved bull spermatozoa. These studies led us to investigate the effects of various thiols, an important class of antioxidants, on sperm motility of cryopreserved bull semen in a commonly used extender, EYTG. Sperm motility was analyzed by computer-assisted semen analysis (CASA). After thawing, a diluted pool of bull semen was incubated at 38.5 degrees C in airtight tubes with the following thiols for 6 hours: glutathione (GSH/GSSG), cysteine, N-acetyl-L-cysteine (NAC) and 2 mercaptoethanol in the presence or absence of oxidative stress. The oxidative stress was caused by adding H2O2 (100 microM) to diluted semen. Incubation of diluted bull semen in EYTG at 38.5 degrees C over a period of 6 h decreased sperm motility by approximately 9 fold from the start (72 +/- 3, mean +/- SEM, n=4) to the end (9 +/- 4, n=4) of the incubation. We found that all thiols to a concentration above 0.5 mM maintained high sperm motility for 6 h in the absence of an external source of oxidative stress (52 +/- 4, for 4 thiols). However, one mM of each thiol was required to efficiently protect sperm motility in the presence of 100 microM of H2O2 for 6 h. We also found that the GSH concentration in diluted semen was too low (microM) to adequately supply exogenous addition of 72 U/mL of glutathione peroxidase (GPx), an enzyme that detoxifies H2O2 and hydroperoxides using GSH as a cofactor. In fact, a better protection of sperm motility could be achieved with only 5 U/mL of GPx and 0.1 mM of GSH added to diluted semen. Our results also demonstrated that added GSSG (0.5 mM) in diluted semen was not regenerated efficiently to GSH over 6 h. The latter result indicated in the extender that the glutathione redox-cycle was deficient. Therefore, deleterious effects sperm motility after cryopreservation in EYTG can be counteracted by adding various thiols at mM concentration. PMID- 11480620 TI - Evaluating recipient and embryo factors that affect pregnancy rates of embryo transfer in beef cattle. AB - The objectives of this experiment were to determine the effects of corpus luteum characteristics, progesterone concentration, donor-recipient synchrony, embryo quality, type, and developmental stage on pregnancy rates after embryo transfer. We synchronized 763 potential recipients for estrus using one of two synchronization protocols: two doses of PGF2alpha (25 mg i.m.) given 11 d apart (Location 1); and, a single norgestomet implant for 7 d with one dose of PGF2alpha (25 mg i.m.) 24 h before implant removal (Location 2). At embryo transfer, ovaries were examined by rectal palpation and ultrasonography. Of the 526 recipients presented for embryo transfer, 122 received a fresh embryo and 326 received a frozen embryo. Pregnancy rates were greater (P < 0.05) with fresh embryos (83%) than frozen-thawed embryos (69%). Pregnancy rates were not affected by embryo grade, embryo stage, donor-recipient synchrony, or the palpated integrity of the CL. Corpus luteum diameter and luteal tissue volume increased as days post-estrus for the recipients increased. However, pregnancy rates did not differ among recipients receiving embryos 6.5 to 8.5 days after estrus (P > 0.1). There was a significant, positive simple correlation between CL diameter or luteal tissue volume and plasma progesterone concentration (r = 0.15, P < 0.01 and r = 0.18, P < 0.01, respectively). There were no significant differences in mean CL diameter, luteal volume or plasma progesterone concentration among recipients that did or did not become pregnant after embryo transfer. We conclude that suitability of a potential embryo transfer recipient is determined by observed estrus and a palpable corpus luteum, regardless of size or quality. PMID- 11480621 TI - Successful direct transfer of vitrified sheep embryos. AB - The use of a simple cryopreservation method, adapted to direct transfer of thawed embryos may help to reduce the costs of embryo transfer in sheep and increase the use of this technique genetic improvement of this species. Two experiments were made to test a vitrification method that is easy to apply in field conditions. All embryos were collected at Day 7 of the estrous cycle of FSH-stimulated donor ewes and were assessed morphologically, washed in modified PBS and incubated for 5 min in 10% glycerol, for 5 min in 10% glycerol and 20% ethylene glycol and were transferred into the vitrification solution (25% glycerol and 25% ethylene glycol). All solutions were based on mPBS. Embryos were loaded in straws (1 cm central part, the remaining parts being filled with 0.8 M galactose in mPBS) and plunged into liquid N2 within 30 sec of contact with the vitrification solution. The straws were thawed (10 sec at 20 degrees C) and the embryos were either transferred directly or after 5 min of incubation in the content of the straw (followed by washing in PBS) into the uterus of a recipient ewe. In Trial 1, the pregnancy rates at term (72 vs. 72%) as well as the embryo survival rates (60 vs 50% respectively) were not different between fresh (n = 48 embryos) and vitrified (n = 50) embryos. In a second trial no difference was observed between vitrified embryos transferred after in vitro removal of the cryoprotectant (n = 86 embryos) or directly after thawing (n = 72) both in terms of lambing rate (67 vs. 75%, respectively) and embryo survival rate (lambs born/embryos transferred; 49 vs. 53%). This method of sheep embryo cryopreservation provided high pregnancy and embryo survival, even after direct transfer of the embryos. PMID- 11480622 TI - Reduction in size of the ovulatory follicle reduces subsequent luteal size and pregnancy rate. AB - We hypothesized that reducing the size of the ovulatory follicle using aspiration and GnRH would reduce the size of the resulting CL, reduce circulating progesterone concentrations, and alter conception rates. Lactating dairy cows (n=52) had synchronized ovulation and AI by treating with GnRH and PGF2alpha as follows: Day -9, GnRH (100 microg); Day -2, PGF2alpha (25 mg); Day 0, GnRH (100 microg); Day 1, AI. Treated cows (aspirated group; n=29) had all follicles > 4 mm in diameter aspirated on Days -5 or -6 in order to start a new follicular wave. Control cows (nonaspirated group: n=23) had no follicle aspiration. The size of follicles and CL were monitored by ultrasonography. The synchronized ovulation rate (ovulation rate to second GnRH injection: 42/52=80.8%) and double ovulation rate of synchronized cows (6/42=14.3%) did not differ (P > 0.05) between groups. Aspiration reduced the size of the ovulatory follicle (P < 0.0001; 11.5 +/- 0.2 vs 14.5 +/- 0.4 mm), and serum estradiol concentrations at second GnRH treatment (P < 0.0002; 2.5 +/- 0.4 vs 5.7 +/- 0.6 pg/mL). The volume of CL was less (P < 0.05) for aspirated than nonaspirated cows on Day 7 (2,862 +/- 228 vs 5,363 +/- 342 mm3) or Day 14 (4,652 +/- 283 vs 6,526 +/- 373 mm3). Similarly, serum progesterone concentrations were less on Day 7 (P < 0.05) and Day 14 (P < 0.10) for aspirated cows. Pregnancy rate per AI for synchronized cows was lower (P < 0.05) for aspirated (3/21=14.3%) than nonaspirated (10/21=47.6%) cows. In conclusion, ovulation of smaller follicles produced lowered fertility possibly because development of smaller CL decreased circulating progesterone concentrations. PMID- 11480623 TI - Preventing experimental vertical transmission of scrapie by embryo transfer. AB - This study investigated whether the transmission of naturally occurring scrapie in sheep can be prevented using embryo transfer. Embryos were collected from 38 donor ewes in a Suffolk sheep flock with a high incidence of naturally occurring scrapie, treated with a sanitary procedure (embryo washing) recommended by the International Embryo Transfer Society and then transferred to 58 scrapie-free recipient ewes. Ninety-four offspring were produced. None of the offspring or the recipient ewes developed scrapie. Furthermore, offspring derived from embryos collected from donor ewes bred to the immunohistochemically positive ram did not develop scrapie. We conclude that scrapie was not transmitted to offspring via the embryo nor was the infective agent transmitted to recipient ewes during embryo transfer procedures. PMID- 11480624 TI - Developmental capacity of equine oocytes matured and cultured in equine trophoblast-conditioned media. AB - The objective was to compare culture media for in vitro maturation of equine oocytes and for in vitro culture of zygotes produced from IVF of partially zona removed oocytes. Cumulus-oocyte complexes from slaughterhouse-derived ovaries were washed in m-Dulbecco's PBS and cultured in TCM-199, F10-DMEM or c-F10-DMEM (50% F10-DMEM + 50% F10-DMEM conditioned medium from culture of an equine trophoblast monolayer for 3 or 4 days). All media included FSH, LH, E2, and 10% FCS. After 28 to 30 h maturation, cumulus expansion was scored from 0 (no expansion) to 4 (fully expanded). Oocytes with a 1st polar body were selected for manipulation after removing cumulus cells using hyaluronidase. About one-third of the zona pellucida was cut using a fragment of a razor blade. For fertilization, fresh stallion semen was washed twice in BGM3 (a modified Tyrode's medium) and capacitated with 0.5 mM c-AMP for 3.5 h and 100 microM ionomycin for 15 min and added to oocytes in fert-TALP at 10(6) spermatozoa/mL. After 20 h, some presumptive zygotes were stained, and the rest were cultured in 100% TCM-DMEM conditioned medium. Cumulus expansion in F10-DMEM and c-F10-DMEM was higher (P<0.05) than the TCM-199 control (3.2, 3.5 vs 1.3, on a scale of 0 to 4). However, polar body formation rates were not different among treatments (47, 52 and 50%). The fertilization rates of equine oocytes matured in TCM-199, F10-DMEM and c-F10-DMEM determined by fixing and staining were 41, 35 and 29%, with no significant differences. There were no significant differences among treatments in cleavage rates (36 to 40%), development to morula (3 to 10%), or blastocyst stages (3 to 5%). On Day 14 of culture in c-F10-DMEM treatment, one blastocyst had more than 500 nuclei, but no capsule was formed. In a further study, cleavage rates (46 to 50%) and development to morula (5 to 10%) and blastocyst stages (3 to 8%) were not different (P>0.1) between TCM-DMEM and 100% conditioned TCM-DMEM for culturing embryos. Six embryos (2 morulae and 4 blastocysts) were nonsurgically transferred to 4 recipient mares, but no pregnancy continued. PMID- 11480625 TI - Detection of estrogen alpha and progesterone receptors and cell proliferation in the uterus during early pregnancy in the goat. AB - The objectives of this study were to investigate in the goat uterus the expression of estrogen-alpha (ER alpha) and progesterone receptors (PR) and their relationship to proliferation indices (Ki-67) during peri-implantation on Days 22 to 30 post coitum (pc). Immunohistochemical methods were used to quantify ER alpha and PR for luminal and deep regions of the endometrium and of the myometrium. On Day 22 pc cell proliferation was only observed in the luminal epithelium. On Day 24 pc, high cell proliferation indices were seen in luminal epithelium and proliferation began in the luminal stroma and glands. There was a positive correlation between Ki-67 and total ER alpha score in the luminal epithelium (r = 0.53, P < 0.01). Levels of PR scores were highly correlated with Ki-67 indices in luminal epithelium (r = 0.74, P < 0.01) and stroma (r = 0.70, P < 0.01). No Ki-67 expression was observed in deep glands, stroma or myometrium on any of the days studied. Results indicate that patterns of ER alpha and PR expression differ markedly, and that there was a high correlation between PR expression and cell proliferation in the caprine uterus during the peri implantation period. PMID- 11480626 TI - Reproductive responses to grazing endophyte-infected tall fescue by postpartum beef cows. AB - The objective was to determine pregnancy rate and stage of embryonic loss in response to grazing endophyte-free (E-; n = 20) or infected (E+; n = 30) tall fescue in postpartum beef cows with calves. Three weeks before estrus synchronization, cow-calf pairs were introduced to pastures (April 1999). Cows were synchronized and bred by AI after detected estrus for a period of 6 d and then by natural service for 62 d. Bulls were rotated weekly to minimize effects of fescue toxicosis on male fertility. Fetal development was monitored weekly between 30 and 60 d of pregnancy and at weaning using transrectal ultrasound. Respiration rate (52.0 +/- 1.4 vs 46.6 breaths/min; P < 0.02) and rectal temperature (39.6 +/- .09 vs 38.8 +/- .12 degrees C; P < 0.001) increased in E+ cows and serum concentrations of prolactin (7.2 vs 57.4 +/- 4.4 ng/mL; P < 0.001), total cholesterol (123.2 vs 149.6 +/- 3.6 mg/dL; P < 0.001), body condition (3.8 vs 5.2 +/- 0.15; P < 0.001; 1 = thin, 9 = fat) and adjusted weaning weight of calves (195.8 vs 210.8 +/- 4.5 kg; P < 0.02) were reduced compared to that of E- cows. Differences were not detected (E- vs E+) for estrus detection rate (84.9 +/- 10.6% vs 80.2 +/- 8.4%), pregnancy rate to synchronized estrus (41.7 +/- 11.8% vs 46.8 +/- 9.5%), overall pregnancy rate 30 d postbreeding (93.8 +/- 6.2% vs 93.5 +/- 5.1%), overall pregnancy rate at 60 d postbreeding (86.7 +/- 10.1% vs 81.2 +/- 8.3%), or serum concentrations of progesterone on day of PGF2alpha treatment (4.5 +/- 0.7 vs 4.5 +/- 0.8 ng/mL). Pregnancy losses that occurred between 30 and 60 d gestation were 6.0 (E-) vs 15.0 (E+) +/- 8.0% (P > 0.10) and occurred after environmental temperatures rose above 37.8 degrees C for three weeks. Total pregnancy losses that occurred by weaning (between 70 and 126 d of gestation) were 5.5 (E-) vs 17.6 (E+) +/- 8.0% (P > 0.10). Pregnancy rate and embryonic losses were not different between cows grazing E- and E+ tall fescue under these management conditions. PMID- 11480627 TI - Oral health during pregnancy: an analysis of information collected by the pregnancy risk assessment monitoring system. AB - BACKGROUND: Little is known about the use of dental services during pregnancy. Yet research suggests that a pregnant woman's oral health and her pregnancy outcome may be associated. METHODS: Four states collected oral health data a part of the Pregnancy Risk Assessment Monitoring System, or PRAMS, in 1998. PRAMS is an ongoing, population-based survey designed to obtain information from mothers who recently delivered live-born infants about their experiences and behaviors before, during and immediately after pregnancy. RESULTS: Reports of dental care use during pregnancy ranged from 22.7 to 34.7 percent. In three states, 12.2 percent to 25.4 percent of respondents reported having a dental problem and of these, 44.7 percent to 54.9 percent went for care. Among mothers reporting a dental problem, prenatal care, or PNC, insurance through public funding and late PNC entry were significantly associated with their not getting dental care. CONCLUSIONS: Most mothers did not go for dental care during their pregnancy; among those who reported having problems, one-half did not get dental care. PRACTICE IMPLICATIONS: Attention toward the oral health needs of pregnant women is warranted. A coordinated effort from the dental and obstetric communities to establish guidelines could benefit maternal oral health and perinatal outcomes. PMID- 11480628 TI - Aiding denture construction: a new twist to an old trick. PMID- 11480629 TI - Effects of audiovisual distraction during dental prophylaxis. AB - BACKGROUND: Fear and anxiety often inhibit patients from seeking dental care. Audiovisual, or A/V, distraction techniques have been shown to reduce patient anxiety and pain during dental procedures. The authors investigated the effects of a virtual image A/V eyeglass system on patients' anxiety and pain. METHODS: Twenty-seven routine dental prophylaxis patients participated and completed the Dental Fear Survey and the Fear of Pain Questionnaire-III before treatment. In random order, the clinician scaled and polished two quadrants in subjects while they watched and listened to a standard video using the A/V eyeglasses and two quadrants while they did not. A posttreatment questionnaire was administered to both the patient and the clinician. RESULTS: Subjects reported less anxiety and discomfort when using the A/V eyeglass system than when they did not. Most subjects preferred to use the A/V equipment rather than receive traditional treatment. The clinician experienced no significant technical interference during the use of the A/V device. The use of the A/V eyeglasses led to decreased treatment time in the first one-half of the procedure. The system appeared to lead to some decreases in the physiological parameters over the course of treatment, with the highest systolic blood pressure occurring after the condition with no use of A/V eyeglasses. CONCLUSIONS: A virtual image A/V system is beneficial in the reduction of fear, pain and procedure time for most dental prophylaxis patients. CLINICAL IMPLICATIONS: Use of screening questionnaires may be helpful for identifying anxious patients. An A/V device may be beneficial to the clinician and the mildly or moderately anxious patient. PMID- 11480630 TI - Self-etching primers are here. PMID- 11480631 TI - High court says appellates should review punitive-damage awards: ruling could curb 'grossly excessive' awards in malpractice cases. PMID- 11480632 TI - For the dental patient. Do you need a mouthguard? PMID- 11480633 TI - Pluses. PMID- 11480634 TI - Third molars. PMID- 11480635 TI - Implant failures. PMID- 11480636 TI - What income sources did you use to finance you dental education? PMID- 11480637 TI - Remember to remember. PMID- 11480638 TI - Accuracy questioned. PMID- 11480639 TI - Tongue cancer rates higher in young adults. PMID- 11480640 TI - Periodontal infection and preterm birth: results of a prospective study. AB - BACKGROUND: Previous studies have suggested that chronic periodontal infection may be associated with preterm births. The authors conducted a prospective study to test for this association. METHODS: A total of 1,313 pregnant women were recruited from the Perinatal Emphasis Research Center at the University of Alabama at Birmingham. Complete periodontal, medical and behavioral assessments were made between 21 and 24 weeks gestation. After delivery, medical records were consulted to determine each infant's gestational age at birth. From these data, the authors calculated relationships between periodontal disease and preterm birth, while adjusting for smoking, parity (the state or fact of having born offspring), race and maternal age. Results were expressed as odds ratios and 95 percent confidence intervals, or CIs. RESULTS: Patients with severe or generalized periodontal disease had adjusted odds ratios (95 percent CI) of 4.45 (2.16-9.18) for preterm delivery (that is, before 37 weeks gestational age). The adjusted odds ratio increased with increasing prematurity to 5.28 (2.05-13.60) before 35 weeks' gestational age and to 7.07 (1.70-27.4) before 32 weeks' gestational age. CONCLUSIONS: The authors' data show an association between the presence of periodontitis at 21 to 24 weeks' gestation and subsequent preterm birth. Further studies are needed to determine whether periodontitis is the cause. CLINICAL IMPLICATIONS: While this large prospective study has shown a significant association between preterm birth and periodontitis at 21 to 24 weeks' gestation, neither it nor other studies to date were designed to determine whether treatment of periodontitis will reduce the risk of preterm birth. Pending an answer to this important question, it remains appropriate to advise expectant mothers about the importance of good oral health. PMID- 11480641 TI - Examining the link between coronary heart disease and the elimination of chronic dental infections. AB - BACKGROUND: While it has been suggested that periodontal disease may be associated with coronary heart disease, or CHD, there are no data to suggest that the elimination of chronic dental infections actually lowers the risk of developing chronic CHD. The goal of this study was to determine whether people with a definitive elimination of all potential dental infections--edentulous people, who are at the optimum endpoint of dental infection elimination from a CHD perspective--lower their CHD risk over time when compared with people who have a specific dental infection, periodontitis. METHODS: The authors examined data from a prospective cohort of 4,027 people who participated in the First National Health and Nutrition Examination Survey, or NHANES I, Epidemiologic Follow-up Study. The primary outcome measure was the first CHD event. RESULTS: During a mean follow-up of 17 years, there were 1,238 CHD events (538 fatal). The confirmed elimination of chronic dental infections did not lead to a decreased risk of experiencing a CHD event (relative risk, 1.02; 95 percent confidence interval, 0.86-1.21). The CHD risk among people with and without chronic dental infections remained constant over time with respect to each other (test for increasing or decreasing trend over time: not significant, chi2(1) = 0.48; P = .93). CONCLUSIONS: People who had a complete, definitive and long-term elimination of all potential dental infections through extraction of all teeth did not have lower CHD risk when compared with people with diagnosed periodontitis. CLINICAL IMPLICATIONS: Until evidence is found to the contrary, the authors suggest that prevention of CHD should not be used as the basis for recommending treatment to eliminate chronic dental infections. PMID- 11480642 TI - Blood pressure outcomes of dental patients screened chronobiologically: a seven year follow-up. AB - BACKGROUND: This article is a follow-up on a 1998 article in JADA. The blood pressure, or BP, of 23 of 24 normotensive patients was monitored at 15-minute intervals for a total of nine days, in three consecutive sessions of four, two and three days, respectively, separated by a few weeks. METHODS: Twelve patients were reached by phone seven years after the prior chronobiologically interpreted monitoring to ascertain their cardiovascular status since the initial monitoring. RESULTS: Only two of the 12 patients reached in follow-up had abnormalities in all three sessions, and only these two patients reported having experienced an adverse vascular event (one a myocardial infarction, the other coronary artery bypass graft surgery). The difference in outcome between the patients with chronobiological abnormality in all three sessions vs. the pool of those with abnormality in only two, one or none of the sessions is statistically significant. CONCLUSIONS: Even a five-day (and sometimes a longer) profile, while greatly preferred to single measurements, may not suffice for a definitive diagnosis of certain patients. Retrospectively, the 864 measurements, on the average, on each person in our study could have served for a recommendation to each person. Chronobiologically interpreted BP and heart rate monitoring for a week or longer as a start detects high-risk states that may be missed by conventional casual measurements that are rarely accompanied by one-day profiles. CLINICAL IMPLICATIONS: The dentist has an important opportunity for conveying the importance of monitoring BP and heart rate for a week to detect an abnormal variability, and for implementing this monitoring. Dentists can educate patients regarding the need for screening and for interpreting the results for variability, and contribute to their overall health by referring them to their physicians when treatment is indicated. PMID- 11480643 TI - Oral lichen planus: patient profile, disease progression and treatment responses. AB - BACKGROUND: Oral lichen planus, or OLP, is a common mucocutaneous immunological disease. The objective of this study was to describe the patient profile, disease progression and treatment responses. METHODS: The authors conducted a retrospective, descriptive study using information from patient records at a tertiary referral center. The study included 229 patients with OLP who were seen in the oral medicine clinic at the University of California, San Francisco, between September 1996 and August 2000, for the first time or for a follow-up visit. Signs and symptoms at various clinic visits were quantified. Responses to treatment and disease progression were determined by comparing scores with baseline scores. RESULTS: The mean age at onset of the disease was 55 years, and 154 (67 percent) of the patients were female. Symptoms generally correlated directly with the severity of OLP forms, which ranged from reticular to erosive. Corticosteroids were effective in reducing symptoms, healing ulcers and reducing erythema. At last follow-up, 65 percent of the patients had the same type of OLP seen initially or the disease had progressed to a more severe type, while 35 percent of patients had less-severe forms than that seen at the initial visit. Four patients (1.7 percent) developed oral squamous-cell carcinoma during the follow-up period. CONCLUSIONS: OLP is a chronic disease with no known cure. Symptoms can improve with corticosteroids; however, the lack of long-term (that is, lifetime) treatment compliance and the adverse side effects of the drugs limit optimal results. CLINICAL IMPLICATIONS: Patients with OLP should be treated if symptoms are significant. Follow-up--including supervision of medication use and monitoring of side effects, as well as periodic examinations for possible malignant transformation--is necessary. PMID- 11480644 TI - Cat-scratch disease: considerations for dentistry. AB - BACKGROUND: Cat-scratch disease, or CSD, results from inoculation of the gram negative bacillus Bartonella henselae via a cat's scratch. A regional lymphadenitis, which usually is cervical, develops and may progress to suppuration. It is necessary to differentiate CSD from other lymphadenopathies. CASE DESCRIPTION: A patient who had close contact with a cat subsequently developed a localized, suppurative cervical lymphadenitis. As B. henselae was identified in 1992, the authors were able to confirm the existence of CSD serologically. Surgical drainage resulted in a successful resolution of the disease process. CLINICAL IMPLICATIONS: As patients with CSD may be seen in the dental office, an awareness of its symptomatology can prevent unnecessary dental intervention and facilitate early treatment. PMID- 11480645 TI - The labiogingival notch: an anatomical variation of clinical importance. AB - BACKGROUND: The labiogingival notch is a developmental phenomenon affecting the maxillary central incisors. Clinically, it can be easily misdiagnosed and improperly treated. Thus, it is important for dentists to recognize this condition. CASE DESCRIPTIONS: The authors describe two cases of misdiagnosed labiogingival notch on a maxillary central incisor. In one case, the notch was misinterpreted as a carious lesion and placement of a restoration was advised. In the second case, external resorption was suspected and forced eruption of the incisor was requested. In both cases, no treatment was needed. CLINICAL IMPLICATIONS: Misdiagnosis of this developmental phenomenon may lead to unnecessary invasive interventions. Because of this, clinicians need to be aware of this clinical variant. PMID- 11480647 TI - Physician, heal thyself: sleep, fatigue, and medical education. PMID- 11480646 TI - The performance of bonded vs. pin-retained complex amalgam restorations: a five year clinical evaluation. AB - BACKGROUND: The authors compared the clinical performance of complex amalgam restorations, replacing at least one cusp--retained either mechanically with self threading pins or bonded--with a filled, 4-methacryloxyethyl trimellitate anhydride, or 4-META-based resin designed for amalgam bonding. METHODS: The authors placed 60 amalgam restorations (28 pin-retained and 32 bonded), each restoration replacing at least one cusp. They used self-threading stainless steel pins in the pin-retained group and a filled, 4-META-based bonding resin in the bonded group. For both groups, the authors left in place any retention form remaining after removal of an old restoration but did not enhance it. RESULTS: At four years, six restorations had failed. At five years, of the 40 restorations available for evaluation, three had failed, for a total of nine failed restorations; seven of those were pin-retained and two were bonded. Using the Fisher exact test to compare the groups at five years, the authors found no significant difference in failure rate, marginal adaptation, marginal discoloration, secondary caries, tooth sensitivity or tooth vitality. CONCLUSIONS: At five years, there was no difference in the performance of pin retained amalgam restorations and bonded amalgam restorations. This study will be continued for at least a sixth year. CLINICAL IMPLICATIONS: Bonding with a filled, 4-META-based bonding resin appears to be a satisfactory method of retaining large amalgam restorations replacing cusps. PMID- 11480648 TI - Spontaneous arousals during quiet sleep in piglets: a visual and wavelet-based analysis. AB - STUDY OBJECTIVES: The objectives of the study were to characterize spontaneous arousals during NREM sleep in piglets and to compare two methods of identifying these events: a "visual" technique using spectral analysis and an automated technique using wavelets. Our goal was to understand the benefits and limits of these methods when applied to sleep in human infants. DESIGN: Arousals were identified by evaluating rapid changes in EEG low frequency activity, blood pressure (BP), and heart rate (HR). A cortical arousal was defined as a rapid decrease in EEG low frequency activity. An autonomic arousal was defined by a transient increase in heart rate or a transient change in mean arterial BP (MAP). SETTING: Laboratory study in sleeping and awake piglets. PARTICIPANTS: Five 1-2 week old piglets. INTERVENTIONS: Chronically instrumented with a femoral arterial line, EEG, EOG, EMG electrodes, and a micro-dialysis probe with its tip located in the rostral ventral medulla. Artificial CSF (aCSF) was dialyzed into the RVM throughout the experiments Measurements: For the visual analysis, the average delta power (0.5-4 Hz) for each 5-second epoch was determined using spectral analysis. MAP and HR were analyzed in 1-second bins. Video images were analyzed for body movements and eye openings. Transient changes in blood pressure, HR, and delta power were then visually identified. For the wavelet analysis, a quantitative, automated technique with a defined "wakefulness threshold" was used to identify rapid decreases in EEG low frequency activity and the rate of change of MAP. RESULTS: Using the visual method, 117 episodes associated with stereotypical hemodynamic, EEG, and behavioral changes (startle) were identified. Seventy five events occurred in isolation or were first in a series of "multiple" events, 41 "multiple" events were defined as events occurring <20 seconds following a previous event. Eighteen events were associated with the termination of apnea. In isolated events or those occurring first in a series, the onset of changes in HR and BP clearly preceded the decrease in EEG amplitude and delta power. Using wavelet analysis, 73 EEG arousals and 115 MAP transients were identified independently; 62% of the EEG events were associated with a transient change in MAP and HR, and in these cases the onset of the hemodynamic events preceded EEG arousals. EEG arousals and MAP transients, however, also occurred alone and not associated with a stereotypical pattern of a startle, changes in MAP and HR and the EEG. CONCLUSIONS: Many of these spontaneous arousals represent integrated EEG, hemodynamic, and behavioral processes similar to arousal phenomena described in adult rats and human infants, but the pattern of spontaneous arousals appears to be more heterogeneous than has been described for arousals induced by exogenous stimuli. Both the visual and wavelet analysis identified these events, but the wavelet technique has the potential advantage of better time resolution and automation of the analysis. PMID- 11480649 TI - The effects of a GABA(A) agonist in the rostral ventral medulla on sleep and breathing in newborn piglets. AB - STUDY OBJECTIVES: Abnormalities in the rostral ventral medulla (RVM) in human infants may contribute to the etiology of the sudden infant death syndrome (SIDS) or a subset of SIDS, by interfering with cardiorespiratory and arousal responses to physiological stimuli often encountered during sleep. The purpose of this study was to determine whether inhibition of groups of neurons in the RVM in newborn piglets would alter sleep and/or the sleep-modulation of breathing. We hypothesized that inhibition of neurons in the RVM would produce less wakefulness or increase the low frequency power (delta) during Quiet sleep. DESIGN: Unanesthetized piglets were studied in a whole-body plethysmograph. Artificial cerebral spinal fluid (aCSF) or the GABAA agonist, muscimol, was dialyzed into the RVM for 40 minutes after a control period consisting of aCSF dialysis. Sleep was analyzed using a combination of EEG spectral analysis and behavioral observations. SETTING: N/A. PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Cardiorespiratory variables varied with state. Dialysis of neither aCSF nor muscimol into the RVM resulted in alterations in resting respiration, BP, HR, or VO2 or their modulation by state. Compared to control dialysis with aCSF, muscimol dialysis caused dramatic effects on sleep architecture. Sleep cycling was abolished in some experiments, whereas in others there were decreases in low-frequency EEG activity or delta power. The animals in which sleep cycling ceased continued in a perpetual state of drowsiness interspersed with periods of wakefulness. CONCLUSIONS: We conclude that dialysis of muscimol into the RVM has little effect on resting breathing, blood pressure, or heart rate or their modulation by state, but interferes with normal sleep architecture. We speculate that abnormalities in the ventral medulla may alter sleep cycling or interfere with arousal mechanisms, thus contributing to the etiology of at least a subset of SIDS. PMID- 11480650 TI - Dental and skeletal changes associated with long-term mandibular advancement. AB - STUDY OBJECTIVES: Little is known of the possible dental or skeletal side effects following the use of mandibular advancement in the treatment of obstructive sleep apnea. A study has subsequently been designed to investigate these issues. DESIGN: 100 consecutively treated medically referred patients were reviewed cephalometrically in 6-month intervals (6-30 months) following mandibular advancement therapy. SETTING: Orthodontic Private Practice. PATIENTS: 87 males, 13 females (mean age 49 years, SD 8.5, range 33-74 years). INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Reference points and planes in the cranial base, maxilla, and mandible were digitized with a reflex metrograph and their means converted to linear and angular measurements. Significant changes following mandibular advancement were observed in lower face height, vertical condylar position, incisor angulation, overbite, and overjet. Skeletal changes were attributed to a vertical repositioning of the mandibular condyle relative to the cranial base and were present at the first review period (6 months). Dental changes occurred later with treatment with the most significant changes occurring at the final review period (30 months) which resulted in a 4.9 degrees proclination of the mandibular incisors and a reduction in overbite of 1.82mm. CONCLUSION: The data suggests that long-term use of mandibular advancement can cause dental and skeletal changes which may be progressive over time. As many consider mandibular advancement a treatment for life, it is strongly recommended that all patients be fully informed of the potential for such changes prior to treatment and undergo mandatory dental reviews with long-term mandibular advancement. PMID- 11480651 TI - Influence of sleep posture on response to oral appliance therapy for sleep apnea syndrome. AB - STUDY OBJECTIVE: This study evaluated the effect of sleep posture on oral appliance therapy to elucidate the interindividual difference of response to the device. DESIGN: Seventy-two unselected patients with sleep apnea syndrome were studied polysomnographically before and after insertion of the individually fabricated and adjusted device. Sleep positions were measured using a body position sensor. The patients were classified into three groups; supine, lateral and prone groups, according to the position in which apneas were most frequently observed. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The mean apnea-hypopnea index (AHI) of all patients before treatment [43.0+/-25.6 (SD)] was significantly (p<0.0001) decreased after insertion of the appliance (21.6+/-18.3). The device decreased the mean AHI significantly from 29.8 to 11.3 in the supine position and 5.5 to 1.6 in the prone position, and increased, but not significantly, from 7.7 to 8.7 in the lateral posture. The supine (n=44) and prone (n=13) groups showed significant reduction of AHI with the oral appliance, while the lateral group (n=15) revealed only a slight decrease, although not significantly. Responders defined by AHI<10 accounted for 61.4% in the supine group, 0% in the lateral group and 84.6% in the prone group. Responders defined by a 50% drop in AHI accounted for 84.1%, 6.7%, and 46.7%, respectively. CONCLUSIONS: The effectiveness of oral appliance therapy is greatly influenced by sleep posture. Sleep posture recorded by polysomnography may be useful to predict the future success or failure of the device. PMID- 11480652 TI - Acute effect of nasal continuous positive airway pressure therapy on the systemic immunity of patients with obstructive sleep apnea syndrome. AB - OBJECTIVES: To investigate whether in patients with obstructive sleep apnea syndrome (OSAS) the systemic immunity is disturbed and whether it changes with nasal continuous positive airway pressure (NCPAP) therapy. DESIGN: Polysomnography was performed on 18 OSAS patients (Group A) before NCPAP was started and again on the first night of NCPAP. Blood samples were collected at 8:00PM, 1:00AM and 6:00AM during each polysomnography. Lymphocyte subsets, lymphocyte blastformation, and natural killer (NK) cell activity were determined. Six normal subjects were also studied. A different six OSAS patients were studied over 6 days of NCPAP. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The only immunological parameter that significantly differed between the Group A OSAS patients either before or on the first night of NCPAP, and the normal subjects was the epinephrine level. Among the Group A OSAS patients, the following immunological parameters were significantly lower at 6:00AM on the first night of NCPAP than before NCPAP was started: percentage (49.4+/-1.9% before NCPAP vs 45.7+/-2.0% with NCPAP, mean+/ SEM, p<0.005) and absolute count of CD4+ cells (944.1+/-63.8 vs 829.6+/-71.3/mm3, p<0.05); absolute count of CD4+HLA-DR+ cells (91.9+/-13.3 vs 75.1+/-8.9/mm3, p<0.05); CD4+/CD8+ ratio (2.13+/-0.21 vs 1.91+/-0.18, p<0.05). The reduction in the percentage of CD4+ cells at 6:00AM was significantly correlated with the change in apnea-hypopnea index (AHI) (r=0.729, p<0.01). The CD4+ cell count recovered after 6 days of NCPAP. The lymphocyte blasfformation and NK cell activity levels did not change with NCPAP. CONCLUSIONS: First-night NCPAP therapy reduced the CD4+ cell count after sleep, which recovered after one week of NCPAP. OSAS patients do not have immunological abnormalities. PMID- 11480653 TI - Effects of a titratable oral appliance on supine airway size in awake non-apneic individuals. AB - STUDY OBJECTIVES: To define morphological changes in the upper airway and its surrounding structures after the insertion of a titratable mandibular repositioner. DESIGN: Ten non-apneic adult males participated in this study. A set of supine lateral cephalograms was taken for each subject at the end of expiration with a titratable oral appliance in place in four mandibular positions: most retruded (RP), maximum protrusion (MAX), 33% of MAX (MAX33), and 67% of MAX (MAX67). Changes in the anteroposterior width of the upper airway, positions of the hyoid bone and the third cervical vertebra were compared between the four mandibular positions. An ANOVA was used to test for statistical significance. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The anteroposterior width of the velopharynx significantly increased when the mandible was advanced from RP to MAX67 and MAX. However, there were no significant changes in the anteroposterior width of the oropharynx. Significant forward displacement of the hyoid bone and third cervical vertebra together with the mandible was found in MAX67 and MAX compared to RP. CONCLUSION: Especially in MAX67 and MAX, the titratable oral appliance significantly enlarges upper airway size in the velopharynx and results in a forward displacement of the hyoid bone and the third cervical vertebra. PMID- 11480654 TI - Age-related increase in awakenings: impaired consolidation of nonREM sleep at all circadian phases. AB - STUDY OBJECTIVES: (1) To assess the circadian and sleep-dependent regulation of the frequency and duration of awakenings in young and older people; (2) to determine whether age-related deterioration of sleep consolidation is related to an increase in the frequency or duration of awakenings; (3) to determine whether pre-awakening sleep structure is preferentially enriched by REM sleep or nonREM sleep and (4) to determine whether sleep structure prior to awakenings is affected by age. DESIGN: Between age-group comparison of sleep consolidation and sleep structure preceding awakenings. SETTING: Environmental Scheduling Facility, General Clinical Research Center. PARTICIPANTS: Eleven healthy young men (21-30 years) and 13 older healthy men (n=9) and women (n=4) (64-74 years). INTERVENTIONS: Forced desynchrony between the sleep-wake cycle and circadian rhythms by scheduling of the rest-activity cycle to 28-h for 21-25 cycles. MEASUREMENTS AND RESULTS: Circadian and sleep-dependent regulation of the frequency and duration of awakenings and of sleep structure preceding awakenings were assessed in 482 sleep episodes (9h 20 min each). The circadian modulation of wakefulness within sleep episodes was primarily related to a variation in the duration of awakenings. In contrast, the age-related reduction of sleep consolidation was primarily related to an increase in the frequency of awakenings. Whereas in both young and older subjects pre-awakening sleep contained more REM sleep than overall sleep, this enrichment of REM sleep (i.e., the gating of wakefulness by REM sleep) was diminished in older people. In older people, preawakening sleep contained more nonREM sleep and stage two sleep in particular, than in young people. CONCLUSIONS: At all circadian phases, the age related reduction of sleep consolidation is primarily related to a reduction in the consolidation of nonREM sleep. PMID- 11480655 TI - Prevalence, clinical characteristics, and risk factors for insomnia in the context of breast cancer. AB - STUDY OBJECTIVES: To estimate the prevalence of insomnia, describe clinical characteristics of sleep difficulties, assess the influence of cancer on the insomnia course, and identify potential risk factors involved in the development of insomnia among women who had received radiotherapy for non metastatic breast cancer. DESIGN: A sample of 300 consecutive women who had been treated with radiotherapy for non metastatic breast cancer first completed an insomnia screening questionnaire. Participants who reported sleep difficulties were subsequently interviewed over the phone to evaluate further the nature, severity, duration, and course of their insomnia. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Nineteen percent (n=56) of the participants met diagnostic criteria for an insomnia syndrome. In most cases (95%), insomnia was chronic. The onset of insomnia followed the breast cancer diagnosis in 33% of the patients and 58% of the patients reported that cancer either caused or aggravated their sleep difficulties. Factors associated with an increased risk for insomnia were sick leave, unemployment, widowhood, lumpectomy, chemotherapy, and a less severe cancer stage at diagnosis. Among women with insomnia symptoms, the risk to meet diagnostic criteria for an insomnia syndrome was higher in those who were separated and had a university degree. CONCLUSIONS: Insomnia is a prevalent and often chronic problem in breast cancer patients. Although it is not always a direct consequence of cancer, pre-existing sleep difficulties are often aggravated by cancer. It is therefore important to better screen breast cancer patients with insomnia and offer them an appropriate treatment. PMID- 11480656 TI - Does cognitive-behavioral insomnia therapy alter dysfunctional beliefs about sleep? AB - STUDY OBJECTIVES: This study was conducted to exam the degree to which cognitive behavioral insomnia therapy (CBT) reduces dysfunctional beliefs about sleep and to determine if such cognitive changes correlate with sleep improvements. DESIGN: The study used a double-blind, placebo-controlled design in which participants were randomized to CBT, progressive muscle relaxation training or a sham behavioral intervention. Each treatment was provided in 6 weekly, 30-60-minute individual therapy sessions. SETTING: The sleep disorders center of a large university medical center. PARTICIPANTS: Seventy-five individuals (ages 40 to 80 years of age) who met strict criteria for persistent primary sleep-maintenance insomnia were enrolled in this trial. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Participants completed the Dysfunctional Beliefs and Attitudes About Sleep (DBAS) Scale, as well as other assessment procedures before treatment, shortly after treatment, and at a six-month follow-up. Items composing a factor analytically derived DBAS short form (DBAS-SF) were then used to compare treatment groups across time points. Results showed CBT produced larger changes on the DBAS-SF than did the other treatments, and these changes endured through the follow-up period. Moreover, these cognitive changes were correlated with improvements noted on both objective and subjective measures of insomnia symptoms, particularly within the CBT group. CONCLUSIONS: CBT is effective for reducing dysfunctional beliefs about sleep and such changes are associated with other positive outcomes in insomnia treatment. PMID- 11480657 TI - Practice parameters for the use of laser-assisted uvulopalatoplasty: an update for 2000. AB - Laser-assisted uvulopalatoplasty (LAUP) is an outpatient surgical procedure which is in use as a treatment for snoring. LAUP also has been used as a treatment for sleep-related breathing disorders, including obstructive sleep apnea. The Standards of Practice Committee of the American Academy of Sleep Medicine reviewed the available literature, and developed these practice parameters as a guide to the appropriate use of this surgery. Adequate controlled studies on the LAUP procedure for sleep-related breathing disorders were not found in peer reviewed journals. This is consistent with findings in the original practice parameters on LAUP published in 1994. The following recommendations are based on the review of the literature: LAUP is not recommended for treatment of sleep related breathing disorders. However, it does appear to be comparable to uvulopalatopharyngoplasty (UPPP) for treatment of snoring. Individuals who are candidates for LAUP as a treatment for snoring should undergo a polysomnographic or cardiorespiratory evaluation for sleep-related breathing disorders prior to LAUP and periodic postoperative evaluations for the development of same. Patients should be informed of the best available information of the risks, benefits, and complications of the procedure. PMID- 11480658 TI - Soybeans and cancer prevention: a complex food and a complex disease. PMID- 11480659 TI - Considering the mechanisms of cancer prevention by selenium. PMID- 11480660 TI - Dietary soy reduces colon carcinogenesis in human and rats. Soy and colon cancer. PMID- 11480661 TI - Selenium metabolism and mechanisms of cancer prevention. PMID- 11480662 TI - Apoptosis and angiogenesis in cancer prevention by selenium. PMID- 11480663 TI - Resveratrol inhibits the expression of cyclooxygenase-2 in mammary epithelial cells. PMID- 11480664 TI - The world of resveratrol. PMID- 11480665 TI - The effects of resveratrol on CYP1A1 expression and aryl hydrocarbon receptor function in vitro. PMID- 11480666 TI - Soy and risk of hormone-related and other cancers. PMID- 11480667 TI - Herbals and cancer. PMID- 11480668 TI - The role of dietary supplements in health. An overview in the United States. PMID- 11480669 TI - The beta-carotene story. PMID- 11480670 TI - Dietary intervention strategies: validity, execution and interpretation of outcomes. PMID- 11480671 TI - Nutrition and cancer prevention: new insights into the role of phytochemicals. Future directions. PMID- 11480672 TI - Chemoprevention: progress and opportunity. PMID- 11480673 TI - Role of tomatoes, tomato products and lycopene in cancer prevention. PMID- 11480674 TI - Tea and tea polyphenols in cancer prevention. PMID- 11480675 TI - Effects of tea polyphenols on the signal transduction pathways. PMID- 11480676 TI - Mechanisms by which garlic and allyl sulfur compounds suppress carcinogen bioactivation. Garlic and carcinogenesis. AB - Overall, a host of studies provides compelling evidence that garlic and its organic allyl sulfur components are effective inhibitors of the cancer process. These studies reveal that the benefits of garlic are not limited to a specific species, to a particular tissue, or to a specific carcinogen. Several mechanisms are likely to account for this protection. Notable among these is a depression in nitrosamine formation and a reduction in carcinogen bioactivation. The benefits provided by garlic must be viewed as part of the entire diet, since several dietary constituents can influence the degree of protection. More than one compound is responsible for the anticancer properties associated with garlic. Future research should focus on how genetic variability and daily environmental factors influence the anticancer benefits attributed to garlic and its allyl sulfur components. PMID- 11480677 TI - Antiproliferative effects of garlic-derived and other allium related compounds. PMID- 11480678 TI - Safety and quality in Australian healthcare: making progress. PMID- 11480679 TI - Non-multiresistant methicillin-resistant Staphylococcus aureus in the community. Next chapter in the story of staphylococcal sepsis? PMID- 11480680 TI - Folate before pregnancy: are we doing enough? PMID- 11480681 TI - Detecting and reducing hospital adverse events: outcomes of the Wimmera clinical risk management program. AB - OBJECTIVES: To determine if an integrated clinical risk management program that detects adverse patient events in a hospital, analyses their risk and takes action can alter the rate of adverse events. DESIGN: Longitudinal survey of adverse patient events over eight years of progressive implementation of the risk management program. PARTICIPANTS AND SETTING: 49,834 inpatients (July 1991 to September 1999) and 20,050 emergency department patients (October 1997 to September 1999) at a rural base hospital in the Wimmera region of Victoria. MAIN OUTCOME MEASURES: Rates of adverse events detected by medical record review and clinical incident and general practitioner reporting. RESULTS: The annual rate of inpatient adverse events decreased between the first and eighth years of the study from 1.35% of all patient discharges (69 events) to 0.74% (49 events) (P<0.001). Absolute risk reduction was 0.61% (95% CI, 0.23%-0.99%), and relative risk reduction was 44.9% (95% CI, 16.9%-72.9%). The quarterly rate of emergency department adverse events decreased between the first and eighth quarters of monitoring from 3.26% of all attendances (84 events) to 0.48% (12 events) (P< 0.001). Absolute risk reduction was 2.78% (95% CI, 2.04%-3.52%), and relative risk reduction was 85.3% (95% CI, 62.7%-100%). CONCLUSIONS: Adverse patient events can be detected, and their frequency reduced, using multiple detection methods and clinical improvement strategies as part of an integrated clinical risk management program. PMID- 11480682 TI - Non-multiresistant and multiresistant methicillin-resistant Staphylococcus aureus in community-acquired infections. AB - OBJECTIVE: To survey Staphylococcus aureus strains isolated from patients presenting from the community, comparing clinical features and antibiotic sensitivity profiles between multiresistant and non-multiresistant methicillin resistant and methicillin-sensitive isolates. DESIGN: Retrospective case series. PARTICIPANTS AND SETTING: Patients who presented to emergency or dermatology departments in hospitals served by the South Western Sydney Area Health Service between 1 May 1998 and 30 April 1999. All patients with methicillin-resistant S. aureus (MRSA) and the first 100 with methicillin-sensitive S. aureus were eligible. MAIN OUTCOME MEASURES: Patient demographic characteristics; risk factors; clinical presentation; treatment; outcome; and isolate antibiotic susceptibility. RESULTS: 139 patients were eligible, and 122 had clinical records available. Ten of these 122 (8%) had multiresistant MRSA, 26 (21%) non multiresistant MRSA and 86 (70%) methicillin-sensitive S. aureus. Among patients with non-multiresistant MRSA, 29% (7/24) were born in New Zealand, Samoa or Tonga, a higher proportion than among those with multiresistant MRSA or methicillin-sensitive S. aureus (P= 0.03). Nearly half (44%) of non multiresistant MRSA strains were community-acquired in patients with no risk factors. Two-thirds of patients with non-multiresistant MRSA (17/26) presented with cellulitis or abscess, and 58% (11/19 evaluable patients) required surgical treatment. CONCLUSIONS: Non-multiresistant MRSA strains are common, especially among people born in New Zealand, Samoa or Tonga, and are usually community acquired. Medical practitioners should routinely swab all staphylococcal lesions for culture and sensitivity. PMID- 11480683 TI - "Folate before pregnancy": the impact on women and health professionals of a population-based health promotion campaign in South Australia. AB - OBJECTIVES: To evaluate a South Australian campaign to promote and implement knowledge that taking adequate folate/folic acid in the periconceptional period can reduce the risk of having a baby with a neural tube defect. DESIGN AND SETTING: The campaign, conducted in October 1994--August 1995, targeted women of reproductive age and health professionals. Evaluation was by computer-assisted telephone interviews undertaken by random dialling throughout the State before and after the campaign, and by self-administered questionnaires to health professionals and women in the postnatal period. PARTICIPANTS: Women of reproductive age and four groups of health professionals. MAIN OUTCOME MEASURES: Knowledge about folate, folate-rich foods and the periconceptional period; participation of health professionals in advising women about folate; use of periconceptional folic acid supplements; sales of folic acid tablets; and prevalence of neural tube defects. RESULTS: Significant increases in knowledge about folate followed the campaign. Health professionals and women in the postnatal period had higher initial levels of knowledge about folate, which also increased significantly. The proportions of women taking periconceptional folic acid supplements, and of health professionals advising women planning a pregnancy about folate, also increased significantly, and folic acid tablet sales doubled. Total prevalence of neural tube defects declined between 1966 and 1999 from a baseline of 2.0 per 1,000 births to 1.1 per 1,000 births (Poisson regression, P= 0.03; average decline of 1.0% per year). CONCLUSIONS: A short educational campaign with a limited budget ($40,000) can promote folate successfully, but alternative strategies such as food fortification are likely to be needed to achieve adequate periconceptional folate intake for a very high proportion of women. PMID- 11480684 TI - Confidentiality in health records: evidence of current performance from a population survey in South Australia. AB - OBJECTIVE: To determine attitudes towards doctors and hospitals as data custodians, and patients' experiences of unauthorised information releases from health services. DESIGN: Analysis of data from a cross-sectional, descriptive household survey (October-November 1999). SETTING: South Australian community. PARTICIPANTS: 3,013 randomly selected residents over 15 years of age. MAIN OUTCOME MEASURES: Level of confidence in doctors and hospitals as data custodians, and patient-reported experience of unauthorised information releases by health services. RESULTS: 288 survey participants (9.6%) were not confident that healthcare providers keep and use information responsibly, 108 (3.6%) reported that healthcare providers had released information without their consent (although at least 48 of these disclosures were legally defensible), and 57 (1.9%) reported harm arising from unauthorised disclosures by health services. Projecting these findings to the South Australian population, over 2,000 people experienced harm arising from unauthorised information release in 1999. However, in the same period, there were fewer than 20 formal complaints to major agencies (eg, Ombudsman, Medical Board). CONCLUSIONS: Healthcare providers have lost the confidence of a minority of patients. For some, this mistrust is based on experience of unauthorised information release. Some disclosures are mandated by legislation. These findings provide baseline performance measures for benchmarking trends in patient confidence and prevalence of unauthorised release of patient information. PMID- 11480685 TI - Healthcare rationing: constraints and equity. AB - It is no longer possible or ethically acceptable to deny that rationing occurs in medical practice. We ration already by using "contraindications to treatment". There are no rationing criteria that are universally ethically acceptable. We need ways to establish community preferences if we are to develop responsible methods of rationing healthcare services. PMID- 11480686 TI - EBM in healthcare: management and policy. AB - The nature of policymaking often precludes a significant role for formal evidence based practice. Management practice is also bereft of formal evidence and appears unlikely to change because of the methodological constraints on collecting good evidence. Despite this, policymakers and managers are keen to promote evidence based clinical practice. This, in part, reflects rational management's desire to standardise the clinical process and develop the profession's accountability to the management hierarchy. To the extent that clinical practice is dependent on organisational settings, this push is inevitable. Widespread and persistent small area variation in clinical practice, and concern over apparent high levels of avoidable error, make doctors vulnerable to these efforts to standardise clinical practice. PMID- 11480687 TI - Truth in clinical research trials involving pharmaceutical sponsorship. AB - Large clinical trials are expensive to mount. Funding comes mainly from pharmaceutical companies seeking information on drug efficacy and adverse events. Patients should be informed of the financial and publication agreements reached between those conducting the trials. This is unlikely to have a significant effect on trial participation and will provide patients with information relevant to informed consent. A small proportion of monies raised from drug trials could be set aside to fund both a trial register site and further studies on adverse drug reactions. PMID- 11480688 TI - Bupropion-induced hypersensitivity reactions. PMID- 11480689 TI - What's causing that gas? PMID- 11480690 TI - Neurological disability and neurological rehabilitation. AB - Evidence is increasing that neurorehabilitation lessens patient disability and improves quality of life in both acute and chronic neurological conditions. A focused, multidisciplinary team approach is the key to a successful rehabilitation outcome. The general practitioner will be more closely involved in the rehabilitation process in the future. Patients will be discharged home earlier to complete the acute rehabilitation program. GPs will supervise function over the long term and activate community rehabilitation resources when necessary to maintain patient function. Ideally, rehabilitation services should be available for most patients with neurological disorders, as it is difficult to predict which individual patients will not benefit. PMID- 11480691 TI - Do magnets alleviate chronic low-back pain? PMID- 11480692 TI - A British epidemic strain of methicillin-resistant Staphylococcus aureus (UK EMRSA-15) in Western Australia. PMID- 11480693 TI - Healthcare workers continue to be at risk of measles: a case for better vaccination coverage. PMID- 11480694 TI - Is osteoporosis undertreated after minimal trauma fracture? PMID- 11480695 TI - Women's access to resources about early breast cancer. PMID- 11480696 TI - Pressure immobilisation bandages in first-aid treatment of jellyfish envenomation: current recommendations reconsidered. PMID- 11480697 TI - Pressure immobilisation bandages in first-aid treatment of jellyfish envenomation: current recommendations reconsidered. PMID- 11480698 TI - Medicine, the media and monetary interests: the need for transparency and professionalism. PMID- 11480699 TI - Will the Royal Australasian College of Physicians Library soon be history? PMID- 11480700 TI - Would the pharmaceutical companies please mind their Ps and Qs, and their Xs, Ys and Zs? PMID- 11480701 TI - Climate change and temperature-dependent biogeography: oxygen limitation of thermal tolerance in animals. AB - Recent years have shown a rise in mean global temperatures and a shift in the geographical distribution of ectothermic animals. For a cause and effect analysis the present paper discusses those physiological processes limiting thermal tolerance. The lower heat tolerance in metazoa compared with unicellular eukaryotes and bacteria suggests that a complex systemic rather than molecular process is limiting in metazoa. Whole-animal aerobic scope appears as the first process limited at low and high temperatures, linked to the progressively insufficient capacity of circulation and ventilation. Oxygen levels in body fluids may decrease, reflecting excessive oxygen demand at high temperatures or insufficient aerobic capacity of mitochondria at low temperatures. Aerobic scope falls at temperatures beyond the thermal optimum and vanishes at low or high critical temperatures when transition to an anaerobic mitochondrial metabolism occurs. The adjustment of mitochondrial densities on top of parallel molecular or membrane adjustments appears crucial for maintaining aerobic scope and for shifting thermal tolerance. In conclusion, the capacity of oxygen delivery matches full aerobic scope only within the thermal optimum. At temperatures outside this range, only time-limited survival is supported by residual aerobic scope, then anaerobic metabolism and finally molecular protection by heat shock proteins and antioxidative defence. In a cause and effect hierarchy, the progressive increase in oxygen limitation at extreme temperatures may even enhance oxidative and denaturation stress. As a corollary, capacity limitations at a complex level of organisation, the oxygen delivery system, define thermal tolerance limits before molecular functions become disturbed. PMID- 11480702 TI - On the evolutionary ecology of host-parasite interactions: addressing the question with regard to bumblebees and their parasites. AB - Over the last decade, there has been a major shift in the study of adaptive patterns and processes towards including the role of host-parasite interactions, informed by concepts from evolutionary ecology. As a consequence, a number of major questions have emerged. For example, how genetics affects host-parasite interactions, whether parasitism selects for offspring diversification, whether parasite virulence is an adaptive trait, and what constrains the use of the host's immune defences. Using bumblebees, Bombus spp, and their parasites as a model system, answers to some of these questions have been found, while at the same time the complexity of the interaction has led expectations away from simple theoretical models. In addition, the results have also led to the unexpected discovery of novel phenomena concerning, for instance, female mating strategies. PMID- 11480703 TI - Evolution and function of auditory systems in insects. AB - While the sensing of substrate vibrations is common among arthropods, the reception of sound pressure waves is an adaptation restricted to insects, which has arisen independently several times in different orders. Wherever studied, tympanal organs were shown to derive from chordotonal precursors, which were modified such that mechanosensitive scolopidia became attached to thin cuticular membranes backed by air-filled tracheal cavities (except in lacewings). The behavioural context in which hearing has evolved has strongly determined the design and properties of the auditory system. Hearing organs which have evolved in the context of predator avoidance are highly sensitive, preferentially in a broad range of ultrasound frequencies, which release rapid escape manoeuvres. Hearing in the context of communication does not only require recognition and discrimination of highly specific song patterns but also their localisation. Typically, the spectrum of the conspecific signals matches the best sensitivity of the receiver. Directionality is achieved by means of sophisticated peripheral structures and is further enhanced by neuronal processing. Side-specific gain control typically allows the insect to encode the loudest signal on each side. The filtered information is transmitted to the brain, where the final steps of pattern recognition and localisation occur. The outputs of such filter networks, modulated or gated by further processes (subsumed by the term motivation), trigger command neurones for specific behaviours. Altogether, the many improvements opportunistically evolved at any stage of acoustic information processing ultimately allow insects to come up with astonishing acoustic performances similar to those achieved by vertebrates. PMID- 11480704 TI - The influence of the physical environment on the self-organised foraging patterns of ants. AB - Among social insects such as ants, scouts that modulate their recruiting behaviour, following simple rules based on local information, generate collective patterns of foraging. Here we demonstrate that features of the abiotic environment, specifically the foraging substrate, may also be influential in the emergence of group-level decisions such as the choice of one foraging path. Experimental data and theoretical analyses show that the collective patterns can arise independently of behavioural changes of individual scouts and can result, through self-organising processes, from the physico-chemical properties of the environment that alter the dynamics of information transfer by chemical trails. PMID- 11480705 TI - Chemical profiles, division of labor and social status in Pachycondyla queens (Hymenoptera: formicidae). AB - Queens of the neotropical ponerine ant Pachycondyla cf. 'inversa' may co-operate during colony founding. One of several co-founding queens specializes in foraging, whereas the others remain in the nest and guard the brood. Division of labor is achieved by aggressive interactions, which result in the formation of dominance hierarchies. Gas chromatography and mass spectrometry of cuticular hydrocarbons obtained from live queens by SPME revealed consistent differences between the patterns of cuticular hydrocarbons of queens with high versus low rank: only high-ranking queens showed considerable amounts of cuticular pentadecane (n-C15) and heptadecene (n-C17:1). These two substances presumably originate from the queens' Dufour glands. PMID- 11480706 TI - Changes in element composition along the spinning duct in a Nephila spider. AB - The silk gland of the golden orb spider Nephila edulis connects to the exit spigot through a long S-shaped duct that assists in the formation of the thread. Previous evidence suggests that the epithelium of the distal (last) part of the duct is specialized for ion transport and that a proton pump is involved in this process. Here, we present evidence from SEM (scanning electron microscope)-EDAX (energy dispersive X-ray) microanalysis of rapidly frozen material maintained at approximately -150 degrees C and from the use of pH indicators that the element composition and pH change progressively as the dragline silk dope (spinning solution) passes down the duct to form the thread. Na+ and Cl- composition decreased while K+ and P and S increased. Indicators suggested that the pH dropped from 6.9+/-0.1 to 6.3+/-0.1. These novel findings suggest that the absorption of Na+ and secretion of the more chaotropic K+ may help the silk protein molecules to refold while the secretion of H+ may assist in this process and reduce the repulsive charges on them. This in turn may allow the molecules to approach one another more closely to crystallize. Thus precise control of the ionic environment within the spider's spinning duct may be important in forming a tough insoluble thread and when devising mimetic processes to spin silk proteins industrially. PMID- 11480707 TI - Cervical cancer screening, screening errors and reporting. AB - Screening for cervical carcinoma by cervicovaginal cytology has led to a marked reduction in the incidence of and mortality from this tumor over the last 50 years in essentially all countries with a functioning screening program. It is the most successful cancer prevention program of all times. Consequently, approximately 80% of the current incidence of and mortality from this disease occurs in geographic areas of underserved and underscreened women. Essential components of a successful program are a high coverage rate of the female population, screening at regular intervals, well-trained clinical and laboratory staff, and an efficient follow-up and treatment system. Deficiencies in any of these areas may lead to a failing screening system. Thus, the most important reason for the remaining mortality from cervical carcinoma in developed countries is lack of complete coverage. It is questionable if new and more expensive technologies will be able to renmedy the remaining failures of the system if no improvement in the coverage rate is achieved. Screening errors do occur but represent only a small fraction of screening failures. Currently, there are a number of terminology systems around the world; thus, a unified terminology is currently not a realistic goal. PMID- 11480708 TI - Summary statement on quantitative cytochemistry (DNA and molecular biology): Task Force 8. AB - OBJECTIVE: To reach consensus on the application of quantitative cytochemical analysis of chromosomal and DNA aneuploidy in cervical cytopathology. CONCLUSION: The current Pap test has limited specificity to predict cancer and its truly progressive pre-malignant lesions. Infection with human papillomavirus may trigger genetic instability, hyperproliferation and immortalization of the cervical mucosa and cause cervical cancer. Several related molecular markers have been shown to be informative about this neoplastic process. Quantitative analysis of chromosomal and DNA aneuploidy has been shown to be an important tool for identifying (progression to) high grade squamous intraepithelial lesions. A high degree of standardization (material handling, calibration and quality control, measurement and interpretation of results) is required for accurate and reproducible measurements. Areas for further study are presented. PMID- 11480709 TI - Obligations to provide appropriate patient management. AB - The Pap test is a successful method of preventing cervical cancer, but it does have significant false negative and false positive rates. The main aim of screening is the detection of precursor lesions, both regression and progression of which may occur, making it difficult to decide upon follow-up and further therapy. Around the world there are many differences, as a far as the frequency of the disease, the organization and economic background of the health care system, the use of different additional diagnostic tools and even the terminology considered. All these factors underline the importance of a consensus on a "minimum level" of obligations to provide appropriate patient management. The screening interval should be two to five years, in some cases even annually. The cytopathologist has an obligation to recommend repeat smears in cases of cytologic abnormalities likely to regress. We recommend the use of standard terminology and stress the importance of a "common language" in cervical cytology. Colposcopy and biopsy are obligatory in cases of HSIL and cancer. We suggest that in severe cases women should be provided with detailed written and verbal information. PMID- 11480710 TI - Cost-effectiveness studies on cervical cancer. AB - Cost-effectiveness analyses are an important source of information for the design and evaluation of policies to reduce cervical cancer. This paper describes the recommendations of a panel on cost-effectiveness studies convened as part of the International Consensus Conference on the Fight Against Cervical Cancer. Recommendations for cost-effectiveness studies include: (1) the use of reference case methods to support comparisons across studies, (2) the use of a consistent standard of evidence on the clinical effectiveness of different screening strategies, (3)further research into the costs and effectiveness of different screening and treatment strategies for cervical cancer, (4) further research into screening and treatment strategies in a wide range of countries, (5) easily accessible and detailed descriptions of the methods and supplementary analyses underlying published studies, (6) greater use of newly developed models of cervical cancer, and (7) greater revelation of potential conflict of interest by researchers. PMID- 11480711 TI - Incidence of cervical carcinoma in the Czech Republic. AB - OBJECTIVE: To analyze the incidence of cervical cancer in the Czech Republic. STUDY DESIGN: Investigation of the cause of the unfavorable level of cervical cancer through incidence and laboratory performance. RESULTS: Repeatedly modified screening program measures did not lower the incidence significantly. The incidence in various regions of the Czech Republic differed without fully correlating with patients' life situations. There has been an important shift in incidence toward teenagers and the elderly. Analysis of laboratory work showed that it is necessary to educate smear takers/clinicians and ensure their cooperation with cytology laboratories. CONCLUSION: The various types of screening programs did not lower the cervical carcinoma incidence. It is essential to provide adequate cytologic smears and sufficient laboratory personnel for their proper evaluation. PMID- 11480712 TI - Accuracy of thin-layer cytology in patients undergoing cervical cone biopsy. AB - OBJECTIVE: To compare the accuracy of thin-layer cytology with Autocyte PREP (TriPath Imaging Inc., Burlington, North Carolina, U.S.A.) with conventional smears in 500 women undergoing cervical cone biopsy. STUDY DESIGN: The study was performed among 500 consecutive women presenting for cone biopsy for high grade cervical intraepithelial neoplasia (CIN) on biopsy in 350 (70%) and discrepant cytology/colpohistology in 150 (30%). Before performing a cone biopsy, two cervical samples were collected for conventional smears and thin-layer cytologic slides, with randomization of the order. Conventional smears were stained and diagnosed at Pasteur Cerba, while thin-layer cytologic slides were processed at a local TriPath office (Meylan, France) and sent in a masked fashion for screening at Pasteur Cerba. Any slides initially read as normal were reviewed again and reported without knowledge of the other cytologic or cone biopsy data. The final cytologic diagnoses for the two methods were compared with histopathology of the cone biopsy. RESULTS: The conventional smear was unsatisfactory in 58 (11.6%) of cases, while there were 4 (0.8%) unsatisfactory thin-layer cytologic slides (P < .001). Endocervical cells were missing from 31 (6.2%) of conventional smears and 34 (6.8%) of thin-layer cytologic slides. For the pooled data, sensitivities of conventional smear and thin layer for detecting high grade CIN (0.82% and 0.86%, respectively) were similar as were specificities (0.40% and 0.43%, respectively). When first samples were compared, the sensitivities of the conventional smear and thin layer for high grade CIN were 0.79% and 0.89%, respectively (P = .02), with corresponding specificities of 0.41% and 0.36% (P < .01). CONCLUSION: When controlled for sample order, the sensitivity of thin-layer cytology for detecting high grade CIN was significantly higher than that of conventional smears in patients with previous abnormal cytology, but at the expense of specificity. PMID- 11480713 TI - Comparing the accuracy of ThinPrep Pap tests and conventional Papanicolaou smears on the basis of the histologic diagnosis: a clinical study of women with cervical abnormalities. AB - OBJECTIVE: To confirm that the ThinPrep Pap test (TP) is as effective as or more effective than the conventional Papanicolaou smear (CS) in detecting epithelial cell abnormalities in a population with cervical abnormalities. STUDY DESIGN: In a blinded, split-sample, matched-pair study, a CS was prepared using a cytobrush, and then TP slides were prepared from the remainder of the sample. All slides were evaluated as defined and classified by the Bethesda System. The results of the two cytologic tests were compared in 483 women relative to the histologic diagnoses of subsequent colposcopically directed cervical biopsies in 158 cases. RESULTS: The cytologic diagnoses from the two methods agreed exactly in 91.4% of cases. The comparison between the two cytologic diagnoses with reference to the histologic diagnosis of subsequent colposcopically directed cervical biopsies showed that TP was significantly more specific for diagnosing lesions than was CS. The sensitivity of the two methods was equivalent. CONCLUSION: In a population with cervical abnormalities, TP is more specific than and as effective as CS in detecting cervical epithelial cell abnormalities. TP improved the specificity of disease detection by reducing the atypical squamous cells of undetermined significance category and/or false positive cases. PMID- 11480714 TI - Is it possible to define a better ASCUS class in cervicovaginal screening? A review of 187 cases. AB - OBJECTIVE: To try to better define the cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS) in a cervical screening protocol. STUDY DESIGN: Smears from 187 patients with cytologic diagnoses of ASCUS and histologic or two years' cytologic/colposcopic follow-up were reviewed. When an ASCUS diagnosis was confirmed, it was done strictly on the basis of the morphologic criteria recommended by the Regione Emilia Romagna Screening Protocol in 1997, trying also to subclassify ASCUS into favor reactive or favor neoplasia. RESULTS: Seventy ASCUS cases were negative (37.4%). Three cases (1.6%) were low grade squamous intraepithelial lesion, and seven (3.8%) were high grade squamous intraepithelial lesion. One hundred seven ASCUS cases (57.2%) were confirmed. Among the 70 negative cases, 36 (51.4%) had reactive changes on biopsy, 30 (42.9%) koilocytosis, 3 cervical intraepithelial neoplasia (CIN 1) and one CIN not otherwise specified (5.7% total). CONCLUSION: Reclassification of ASCUS cases using tighter criteria reduced them to a rate of 57.2% but missed 30 patients with histologic diagnoses of koilocytosis and 4 with histologic diagnoses of CIN. PMID- 11480715 TI - Fine needle aspiration cytology of metastatic scalp nodules. AB - OBJECTIVE: To analyze cases of palpable cutaneous/ subcutaneous scalp nodules and evaluate the clinical features and role of fine needle aspiration cytology (FNAC) in their diagnosis. STUDY DESIGN: A total of 1,022 cases of metastatic malignancies at various sites were analyzed over a period of two years. FNAC was performed as a routine outpatient procedure for palpable lesions and under ultrasound or computed tomographic guidance for nonpalpable, internal sites. Histologic correlation was done when possible. RESULTS: Eighteen cases were found to be metastatic scalp nodules. The scalp as a metastatic site constituted 1.76% of all metastatic sites, 7.72% of extranodal sites, and 15.3% of extranodal and extrahepatic sites. Of the 18 cases, 7 (38.9%) had primaries in the head and neck region (thyroid, 3; orbit, 1; tonsil, 1; tongue, 1; and parathyroid, 1). Cutaneous/subcutaneous metastasis was encountered in 58 patients, and the 18 cases of scalp metastasis constituted 31.03% of all cutaneous sites. CONCLUSION: Careful examination of the skin on the entire body, with particular attention to the scalp, should be done when a primary internal malignancy or recurrence is suspected. FNAC is a very helpful and cost-effective modality in determining the nature of such lesions. PMID- 11480716 TI - Cytologic findings of medulloblastoma in crush smears. AB - OBJECTIVE: To describe the cytologic findings of medulloblastomas on intraoperative crush preparation smears and compare them with the findings of other central nervous system tumors. STUDY DESIGN: The intraoperative crush preparation smears of 19 cases of medulloblastoma were studied (6 undifferentiated type and 13 well-differentiated type). The findings were compared with those of a control group consisting of 31 astrocytomas, 22 ependymomas, 18 oligodendrogliomas, 27 meningiomas, 17 schwannomas, 17 pituitary adenomas, 3 lymphomas, 5 hemangioblastomas, 5 chordomas and 11 metastatic tumors. RESULTS: Medulloblastomas revealed clusters and isolated small, round malignant cells with hyperchromatic nuclei and indistinct cytoplasm. Typical Homer-Wright rosettes were seen in the well-differentiated type, but they were poorly formed in the undifferentiated type. Tumor cannibalism, target inclusions, cytoplasmic vacuoles and prominent multiple nucleoli were noticed frequently in the undifferentiated type. The control group (metastatic tumors and high grade astrocytomas) rarely showed tumor cannibalism or multiple nucleoli. CONCLUSION: Smears of the undifferentiated type of medulloblastomas frequently revealed tumor cannibalism, cytoplasmic vacuoles, target inclusions and prominent multiple nucleoli. These findings have been rarely reported. The prognosis of the undifferentiated type of medulloblastoma was poor. PMID- 11480717 TI - Subclassification of localized Leishmania lymphadenitis in fine needle aspiration smears. AB - OBJECTIVE: To describe the cytologic findings of localized Leishmania lymphadenitis and discuss the differential diagnosis. STUDY DESIGN: The study group consisted of 133 cases. All of them were diagnosed by fine needle aspiration (FNA) study. The ages ranged between 3 and 80 years, 102 were male and 31 female. Seventy lymph nodes were excised. RESULTS: The FNA smears revealed a polymorphic population of cells composed of lymphocytes, histiocytes, giant cells, abnormal plasma cells and tingible body macrophages. Leishman-Donovan (LD) bodies were identified in all cases, but their number differed from case to case. Granulomas, dendritic cells, mast cells and lymphoglandular bodies were identified in a substantial number of cases. Depending upon the presence of characteristic cytologic findings, the cases were divided into five major groups: acute inflammation with giant cells, histiocytic granulomas, epithelioid cell granulomas, plasma cell type and mixed histioplasmacytic type. CONCLUSION: Leishmaniasis is an uncommon cause of cervical lymphadenitis but should be considered in the differential diagnosis of unexplained lymphadenopathy in endemic countries. Demonstration of LD bodies is necessary for the diagnosis of this self-limited condition, for which no treatment is required. PMID- 11480718 TI - Pap smears in women with endometrial carcinoma. AB - OBJECTIVE: To correlate Pap smear findings with the histology of endometrial carcinoma and stage of the disease. STUDY DESIGN: Between 1995 and 1998, 76 women with endometrial carcinoma, having had Pap smears done within two to three months of hysterectomy at Memorial Sloan-Kettering Cancer Center, formed the basis for this study. All Pap smears and histologic sections were reviewed. RESULTS: Thirty four patients had normal Pap smears (45%), and 42 had abnormal ones (55%). The mean age of the two groups was 65.1 and 65.2 years, respectively. Histologic subtypes included 44 International Federation of Gynecology and Obstetrics (FIGO) grade 1 endometrioid adenocarcinoma (low grade) and 32 high grade carcinomas, including 19 FIGO grade 2 or 3 endometrioid adenocarcinomas, 5 papillary serous carcinomas (PSC), 2 clear cell carcinomas (CC), 1 adenosquamous carcinoma, 3 endometrioid adenocarcinomas mixed with PSC and 2 endometrioid adenocarcinomas mixed with CC. The proportions of patients with low and high grade tumors with abnormal Pap smears were 43% (19/44) and 72% (23/32), respectively (P=.01). The proportions of patients with abnormal Pap smears and no myometrial invasion, invasion of <50% and >50% myometrial thickness were 40% (8/20), 62% (26/42) and 57% (8/14), respectively (P =.27). Vascular invasion was identified in 56% (9/16) of patients with abnormal Pap smears and in 55% (33/60) of patients with normal ones (P = .93). The proportions of patients having abnormal Pap smears with stage I and stages II, III or IV disease were 48% (30/62) and 86% (12/14), respectively (P =.01). CONCLUSION: Although the Pap smear is not a sensitive screening test for endometrial cancer and a negative Pap smear does not rule it out, this study revealed that abnormal Pap smears are significantly associated with high grade of tumor and stage II-IV endometrial carcinoma. However, they are not associated with patient age, depth of myometrial invasion or vascular invasion. PMID- 11480719 TI - Malignant pericardial effusion. AB - OBJECTIVE: To examine the frequency and diagnostic implications of malignant pericardial effusions. STUDY DESIGN: The clinical and histologic records of pericardial effusions diagnosed cytologically as malignant were reviewed. We investigated the relationship between malignancy and type of tumor, interval between diagnosis of the primary tumor and development of the pericardial effusion, and length of survival after the onset of the effusion. RESULTS: There were 375 pericardial effusions among 23,592 effusions studied over 24 years; 65 of them were diagnosed as malignant. The mean age at onset was 53.6 years. In 92% of the cases the primary tumor was epithelial, lung the most frequent in males and breast in females. In 48% of cases the pericardial effusion constituted the first sign of malignancy. CONCLUSION: Diagnosis of a pericardial effusion as malignant carried an ominous prognosis: 86% of patients died within the first year and nearly one-third within the first month. Breast carcinomas were linked with the longest survival and longest interval of latency prior to the onset of the pericardial effusion. There were two effusions associated with lymphoblastic lymphomas; they have had no recurrence 10 and 17 years after the effusion, and the patients can be considered cured. PMID- 11480720 TI - Salivary gland fine needle aspiration using the ThinPrep technique: diagnostic accuracy, cytologic artifacts and pitfalls. AB - OBJECTIVE: To retrospectively assess the diagnostic accuracy, cytologic features and pitfalls of ThinPrep (TP) (Cytyc Corporation, Marlborough, Massachusetts, U.S.A.) versus conventional (smear) preparation (CP) in salivary gland fine needle aspiration biopsies (FNABs) and second, to evaluate the reproducibility of the cytomorphologic criteria used in the evaluation of FNABs prepared by CP versus TP. STUDY DESIGN: All salivary gland fine needle aspiration biopsies (SGFNABs) between January 1996 and June 1999 were retrieved from the cytology files of the University of Michigan Hospital. Histologic correlation was identified when available. Two cytopathologists reevaluated the slides for artifacts, cellular preservation, background material, cellularity, and cytoplasmic and nuclear details. RESULTS: Seventy-four of the 134 (55%) cases identified had histologic follow-up. Fifty (68%) cases were processed by TP and 24 (32%) by CP. FNAB processed by TP and CP correctly identified malignancy in 14 and 9 cases, respectively. There were three (4%) false negative cases. These included two acinic cell carcinomas and one mucoepidermoid carcinoma. There were 37 true negative cases (24 TP and 13 CP) and one false positive case of cellular pleomorphic adenoma (cytologic interpretation, mucoepidermoid carcinoma). All discrepant cases were processed using the TP method. The overall specificity and sensitivity were 98% and 88%, respectively. However, specificity and sensitivity for TP-processed SGFNABs were 96% and 82% as compared to a 100% specificity and sensitivity for CP. Additionally, there were 10 (14%) nondiagnostic cases, 8 of which were processed by TP. Cytologic artifacts associated with TP included diminished/distorted extracellular and stromal elements, cellular shrinkage and tissue fragmentation CONCLUSION: The diagnostic accuracy of TP-processed SGFNABs approaches that of the CP. However, there are several artifacts that may lead to erroneous diagnoses. Additional studies, that depend on real-life clinical samples processed by TP are suggested to modify current diagnostic criteria. PMID- 11480721 TI - Tumor markers in the detection of recurrent transitional cell carcinoma of the bladder: a brief review. AB - OBJECTIVE: To determine the status of the most promising tumor markers of bladder cancer, including comparison with cytology, technical complexity and utility in patient management. STUDY DESIGN: An extensive literature search was performed, and multiple markers were evaluated. The markers with the greatest potential for use as an adjunct to cytology were reviewed to determine the value of clinical implementation. Markers with a paucity of clinical research and poor results in clinical trials were omitted from review, as were genetic and cytologic prognostic determinants. RESULTS: NMP22, bladder tumor antigen, fibrin/fibrinogen degradation products, telomerase and QUANTICYT image analysis cytometry produced the most favorable and reproducible results. Each test obtained favorable sensitivities in comparison with cytology, especially in the detection of low grade lesions. Many also retrospectively placed patients in high- and low-risk groups based on the test results, allowing increased follow-up time between cystoscopies. However, inability to detect some high grade lesions reduces their utility. CONCLUSION: Continued clinical trials using these and other predictors of bladder cancer will eventually find a test that is suitable, in sensitivity and specificity, for use in urology clinics. Until that time, these tests may be useful in conjunction with cytology to prolong the interval between cystoscopies. PMID- 11480722 TI - Cytodiagnosis of simple and proliferating trichilemmal cysts. AB - OBJECTIVE: To elucidate the cytologic findings in simple trichilemmal (pilar) cysts and proliferating trichilemmal cysts (pilar tumors) and the clinical importance of these lesions. STUDY DESIGN: Aspirates from 12 simple pilar cysts and three pilar tumors, all histologically confirmed, were analyzed with a view to elaborating on specific cytologic features enabling a distinction from epidermal cysts and other adnexal tumors. RESULTS: Aspirates from pilar cysts showed two different pictures, depending on the age of the cyst. Young pilar cysts showed aspirates with an abundant background of blotchy keratin with or without calcification and inflammation. Older (degenerating) cysts showed oily fluid debris with cholesterol crystals and inflammatory cells. The epithelial component was sparse, and only an occasional syncitial cluster of small squamoid cells was seen. As opposed to this, epidermal cysts usually showed a cleaner background, with very cellular aspirates containing many nucleate and anucleate squames, keratin flakes, platelike crystals and no calcification. The pilar tumors showed a lesser amount of keratin as compared to pilar cysts and yielded large and small clusters of squamoid and basaloid cells, a few of which showed an abrupt association with anucleate, keratinized globules. CONCLUSION: The cytologic diagnosis of a pilar cyst should be made especially in scalp cysts, which yield either abundant, blotchy keratin or oily, cholesterol-rich debris with a sparse epithelial component and which lack a mixture of anucleate and nucleate squames. On aspiration, pilar tumors yield comparatively more cells and less keratin and show small, basaloid or squamoid cells abruptly associated with keratin globules. The cytologic diagnosis of pilar cysts is important because these cysts recur if incompletely excised and often undergo transformation to pilar tumors. Similarly, pilar tumors often clinically mimic squamous carcinomas, and hence there is a need to outline definitive cytologic features. PMID- 11480723 TI - Fine needle aspiration cytology in lymphadenopathy of HIV-positive cases. AB - OBJECTIVE: To evaluate the role of fine needle aspiration biopsy (FNAB) material in 25 HIV-positive cases with lymphadenopathy. STUDY DESIGN: We selected 25 cases for the present study who were enzyme-linked immunosorbent assay positive for HIV (HIV-1). FNAB was performed as a routine, outdoor procedure with informed consent of the patient. For each case, along with routine May-Grunwald-Giemsa and hematoxylin and eosin staining, Ziehl-Neelsen staining for acid-fast bacilli and periodic acid-Schiff staining for fungi were performed wherever necessary. RESULTS: A total of 28 sites were aspirated from 25 HIV patients. All these patients were heterosexual, and none had a history of drug abuse. FNAB was performed under ultrasound guidance in all four cases of a retroperitoneal group of lymph nodes. The most common FNAB diagnosis was reactive lymphoid hyperplasia (10), followed by tuberculosis (8). There were three cases diagnosed as fungal infection (two, Cryptococcus; one, histoplasmosis). FNAB of a case of lymph node was suggestive of tuberculosis. There was one case each diagnosed as non Hodgkin's lymphoma and squamous cell carcinoma (metastatic). One case of a small axillary lymph node did not yield representative material. CONCLUSION: FNAB is a relatively inexpensive initial investigative technique in the diagnosis and management of HIV-positive patients. It can obviate the need for surgical excision and enable immediate treatment of specific infections. PMID- 11480724 TI - Fine needle aspiration cytology of invasive cribriform carcinoma of the breast with osteoclastlike giant cells: a case report. AB - BACKGROUND: Nonneoplastic osteoclastlike giant cells are occasionally associated with carcinoma of the breast, pancreatobiliary and gastrointestinal systems. In the breast, this uncommon stromal response is seen mainly in invasive carcinoma with low grade cytology, among which invasive cribriform carcinoma is the classic example. Details of the fine needle aspiration cytology of this phenomenon, especially in thin-layer preparations, have been described rarely. CASE: The fine needle aspiration cytology of an invasive cribriform carcinoma of the breast occurred in a 66-year-old woman. Cytology showed cohesive sheets and three dimensional cribriform clusters of bland-looking and mitotically inactive ductal cells in a blood-stained background. Scattered multinucleated, osteoclastlike giant cells, some containing hemosiderin granules, were also seen. Myoepithelial cells and naked nuclei were not obvious. The cellular composition was more discernible in liquid-based cytologic preparations. Histologic examination of the excisional biopsy showed an invasive cribriform carcinoma associated with many osteoclastlike giant cells in a hypervascular stroma. CONCLUSION: In view of the extremely low grade cytology of the malignant ductal cells, invasive cribriform carcinoma may closely mimic benign proliferative breast diseases on fine needle aspiration biopsy. Recognition of this special relationship with osteoclastlike giant cells, which are rarely present in certain subtypes of breast cancer but not benign lesions, can help to arrive at a correct cytologic diagnosis. PMID- 11480725 TI - Micropapillary variant of transitional cell carcinoma of the urinary bladder: a report of three cases with cytologic diagnosis in urine specimens. AB - BACKGROUND: Micropapillary transitional cell carcinoma is a recently described, aggressive variant of bladder cancer. Its cytologic features in urine have not been previously characterized. CASES: Three cases illustrate the urinary cytologic features of this high grade urothelial carcinoma and its concurrent biopsy findings. This tumor is similar to low. grade urothelial lesions of the bladder, tends to present as micropapillary clusters in urine and yet has high grade nuclear features within these clusters that help with the differential diagnosis of a flat, high grade urothelial carcinoma. CONCLUSION: The micropapillary type of transitional cell carcinoma is a distinct morphologic entity with an aggressive clinical course. Recognizing its presence in urinary cytology, albeit a rare occurrence, is important in distinguishing this lesion from the more indolent, low grade papillary lesions and high grade urothelial carcinomas, which continuously shed single malignant urothelial cells. PMID- 11480726 TI - Mucinous cystadenocarcinoma of the parotid gland: report of a case with fine needle aspiration findings and histologic correlation. AB - BACKGROUND: Mucinous cystadenocarcinoma of the salivary gland is a rare entity. Review of the literature from 1991 to 1999 revealed no previous reports on its cytologic features. CASE: A 25-year-old man had a slowly growing, painless mass in the left parotid gland. Fine needle aspiration biopsy, performed prior to surgical excision, showed clusters of minimally atypical epithelial cells in which occasional vacuolated cells containing mucin could be seen. Pathologic evaluation of the resected parotid mass showed it to be a mucinous cystadenocarcinoma. CONCLUSION: The cytologic differential diagnosis of mucinous cystadenocarcinoma is with low grade mucoepidermoid carcinoma and with mucinous adenocarcinoma. Mucinous cystadenocarcinoma must be cystic; cysts may be present in low grade mucoepidermoid carcinoma, but their size and prominence varies. Mucinous adenocarcinoma is not cystic but gelatinous. Nuclei are bland in both mucinous cystadenocarcinoma and low grade mucoepidermoid carcinoma but are atypical in mucinous adenocarcinoma. There is no squamous differentiation in either mucinous cystadenocarcinoma or mucinous adenocarcinoma, but it is subtle in low grade mucoepidermoid carcinomas. Mucinous cystadenocarcinoma should be considered a potential candidate in the differential diagnosis of mucinous lesions that can occur in the salivary gland. PMID- 11480727 TI - Transient myeloproliferative disorder in Down syndrome presenting with ascites: a case report. AB - BACKGROUND: An increased frequency of acute myelogenous leukemia is a well known feature in children with Down syndrome. In addition, transient myeloproliferative disorders (TMD), which may mimic acute leukemia, also occur in neonates with Down syndrome. TMD is recognized shortly after birth or in the neonatal period and is characterized by leukocytosis and thrombocytopenia, which resolve spontaneously in four to six weeks. CASE: A 1.5-month-old, male infant born with Down syndrome and patent ductus arteriosus presented with abdominal distention due to ascites. Cytology of the fluid revealed immature myeloid cells and megakaryocytes. Flow cytometry of the ascitic fluid confirmed the presence of immature myelomonocytic cells. A complete hematologic evaluation along with the clinical findings supported the diagnosis of TMD in Down syndrome. CONCLUSION: TMD is an uncommon syndrome strongly associated with Down syndrome. Since the abnormal laboratory findings are seen primarily in the peripheral blood, it is usually diagnosed by a hematopathologist without much difficulty. Our case demonstrates the importance of cytopathologist familiarity with this entity so as not to erroneously diagnose a leukemic process. This is extremely important since most cases of TMD spontaneously resolve within a few weeks to months and do not require treatment other than supportive measures. PMID- 11480728 TI - Mullerian adenosarcoma of the uterus: report of a case with imprint cytology. AB - BACKGROUND: Mullerian adenosarcoma is a rare morphologic variant of uterine sarcoma that, although well described histologically, is scarcely mentioned in the cytologic literature. CASE: A 75-year-old female was suspected of having atypical endometrial hyperplasia on an endometrial smear. However, subsequent imaging techniques revealed the presence of a bulky, polypoid mass filling the uterine cavity. On pathologic examination of the hysterectomy specimen, the polypoid tumor was diagnosed as mullerian adenosarcoma, homologous, with sarcomatous overgrowth, in which the sarcomatous component was compatible with high grade endometrial stromal sarcoma. Imprint smears of the tumor consisted of two morphologic patterns, sarcomatous and glandular. The sarcomatous tumor cells, with coarse chromatin and relatively scant cytoplasm, formed small aggregates or appeared alone. These cells were semiround or oval and had conspicuous nucleoli. In addition to these observations, small and large clusters of glandular cells with mild atypism were interspersed with the sarcomatous cells. CONCLUSION: Cytologic examination of mullerian adenosarcoma well reflects its pathologic features. PMID- 11480729 TI - Intraabdominal desmoplastic small round cell tumor: report of a case with fine needle aspiration, cytologic diagnosis and molecular confirmation. AB - BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a recently described neoplasm. This entity is well defined, with distinct clinical, pathologic and immunohistochemical features. Molecular studies have shown a specific reciprocal translocation t(11;22)(p13;q12). To our knowledge, no report of DSRCT with molecular confirmation on cytologic material has been reported before. CASE: Fine needle aspiration (FNA) was performed on an intraabdominal mass in a 37-year-old man. A May-Grunwald-Giemsa-stained preparation showed clusters of small round tumor cells associated with desmoplastic stromal cells, highly suggestive of DSRCT. FNA of a supraclavicular node showed cytologic features similar to those of the primary abdominal mass, including a prominent desmoplastic reaction of the stroma. Immunocytochemical studies showed myogenic and epithelial differentiation. Molecular analysis was performed on FNA, revealing the EWS/ WT1 chimeric transcript and thus confirming the cytologic diagnosis. CONCLUSION: Cytomorphologically, a definitive diagnosis of DSRCT may be difficult, as this tumor bears considerable resemblance to other small round cell tumors. The diagnosis can be confirmed by ancillary techniques, such as immunocytochemistry, and particularly by molecular analysis, which may also be performed on cytologic material. PMID- 11480730 TI - Duodenal somatostatinoma of the ampulla of vater diagnosed by endoscopic fine needle aspiration biopsy: a case report. AB - BACKGROUND: Duodenal somatostatinoma is a rare neuroendocrine neoplasm. A better prognosis can be obtained if these tumors are resected at an early clinical stage. Endoscopic punch biopsy has been the method most commonly used for the preoperative diagnosis of neuroendocrine duodenal tumors. To the best of our knowledge, endoscopic fine needle aspiration biopsy (FNAB) of duodenal somatostatinoma has not been reported before. CASE: A 41-year-old, black female presented with upper gastrointestinal bleeding as well as elevated bilirubin, liver enzymes and glucose. Computed tomography, esophagogastroduodenoscopy and endoscopic retrograde cholangiopancreatography (ERCP) detected a mass at the region of the ampulla of Vater partially obstructing the pancreatic duct. The initial punch biopsy yielded only intestinal mucosa. Subsequent endoscopic FNAB suggested the diagnosis of a neuroendocrine neoplasm, as confirmed by additional punch biopsies. Immunohistochemical and electron microscopic studies disclosed somatostatin production by the tumor, which was resected through a modified Whipple procedure. The patient recovered fully. CONCLUSION: This case demonstrates the usefulness of endoscopic FNAB in diagnosing submucosal gastrointestinal neuroendocrine tumors. PMID- 11480731 TI - Arias-Stella atypia in a paratubal cyst: a rare phenomenon with potential diagnostic pitfalls. AB - BACKGROUND: Arias-Stella reaction is commonly encountered in the endometrium during the gestational period and puerperium. Similar changes in the tubal epithelium are exceedingly rare. The cytologic findings of this phenomenon have never been described before. CASE: Cytologic findings of Arias-Stella changes in the lining cells of a left paratubal cyst in a 41-year-old woman were examined. The smears showed occasional small clusters or isolated tubal epithelial cells consisting of a mixture of ciliated and secretory cells. Some of the ciliated cells showed a marked increase in nuclear and cytoplasmic volume and possessed coarse chromatin, prominent, angulated nucleoli, dense, amphophilic cytoplasm; and a discrete cell membrane. The nuclear membrane was still thin and regular, and these cells were mitotically inactive. The preservation of apical cilia in some of them ensured their benign nature. In some areas, clusters of secretory cells with an increase in both nuclear and cytoplasmic volume and ample, clear cytoplasm were also found. There were small, monolayered sheets of regular and benign-looking epithelial cells with a honeycomb arrangement, suggesting that the Arias-Stella atypia was focal. CONCLUSION: In view of the presence of scattered giant cells with coarse chromatin and angulated macronucleoli, a false positive cytologic diagnosis of malignancy can be made if one does not pay attention to the coexistence of apical cilia and other subtle changes, including the usual nuclear/cytoplasmic ratio, thin and regular nuclear membrane and absence of mitotic activity. PMID- 11480732 TI - Primary malignant melanoma of the urinary bladder diagnosed by urine cytology: a case report. AB - BACKGROUND: Primary melanoma of the urinary bladder is a rare neoplasm, and there have been no prior reports in which the initial diagnosis was made by urinary cytology. CASE: An 82-year-old woman presented with vaginal spotting, gross hematuria and dysuria. Voided urine cytology revealed malignant cells, several of which exhibited cytoplasmic melanin pigment and were accompanied by many macrophages also containing melanin. Cystoscopy revealed a darkly pigmented, polypoid mass at the bladder neck. Biopsy confirmed the diagnosis. CONCLUSION: Primary melanoma of the urinary bladder is rare. The diagnosis can be made on cytologic examination of voided urine if careful study of exfoliated malignant cells reveals cytoplasmic melanin pigment. Macrophages may also harbor melanin pigment, and their presence should alert the cytopathologist to search carefully for pigmented malignant cells. Clinical and radiologic studies are essential to rule out melanoma metastatic to the bladder. PMID- 11480733 TI - Crush and imprint cytology of subependymoma: a case report. AB - BACKGROUND: There are few descriptions of the cytologic features of subependymoma because this neoplasm is rare and most commonly encountered incidentally at autopsy. Here we report a surgical case of subependymoma occurring in the lateral ventricle and provide the first documentation of the crush cytologic features of this tumor. CASE: A 34-year-old woman was found to have a tumorous lesion in the right lateral ventricle. At surgery, a 2-cm-diameter tumor was detected in the anterior horn. Histologic examination during surgery revealed that the mass was composed of loose,fibrillary networks and clusters of nuclei showing mild pleomorphism. A number of microcystic formations were evident. Histologically, the neoplasm was considered benign--specifically, a subependymoma. Papanicolaou- or Giemsa-stained crush specimens and imprint smears were also prepared. The cytologic morphology was fundamentally the same as the histologic. In the crush specimens, microcystic formations were readily visible. Moreover, details of the cellular morphology were more easily recognized in the cytologic slides than in the frozen sections. CONCLUSION: Cytologic examination, particularly crush cytology, appears to be useful for the rapid diagnosis of subependymoma during surgery in combination with the examination of frozen histologic sections. PMID- 11480734 TI - Fine needle aspiration cytology of a dedifferentiated liposarcoma: report of a case with histologic and immunohistochemical follow-up. AB - BACKGROUND: Dedifferentiation is a histologic progression of a neoplasm from low grade to high grade histology. It occurs in tumors of the retroperitoneum and in those undergoing treatment. This usually occurs in the setting of radiation or chemotherapy or as a spontaneous process over a long period. The features of dedifferentiation can be toward any mesenchymal element of the underlying neoplastic process. CASE: We report the cytologic features of a dedifferentiated liposarcoma arising in a 76-year-old man who had a history of well-differentiated liposarcoma. Papanicolaou- and Diff-Quik-stained smears from a radiologically guided fine needle aspiration biopsy showed a hypercellular sample. The smears showed a mixed population of cells. There were multinucleated, pleomorphic giant cells with abundant cytoplasm, smaller clusters of cells with a high nuclear/cytoplasmic ratio and cells with spindled and elongated nuclear features. The follow-up surgical resection specimen showed a dedifferentiated liposarcoma with strong and diffuse immunoreactivity to vimentin, desmin and CD68 in the large, pleomorphic cells; focal and weak immunoreactivity to smooth muscle actin and S-100 in these cells; and strong and focal immunoreactivity to desmin, smooth muscle actin and muscle-specific actin in the spindle cells. This supports the dedifferentiated components of this tumor to be of fibrohistiocytic and leiomyosarcomatous differentiation. CONCLUSION: Dedifferentiation of a well differentiated liposarcoma should be entertained in the setting of a mass lesion in the retroperitoneum in patients with prior histories of well-differentiated liposarcoma. The radiologic features of a particular neoplastic process can be very helpful in determining the nature of this process. PMID- 11480735 TI - Vorticella in Pap smears as a contaminant. PMID- 11480736 TI - Fine needle aspiration diagnosis of bronchogenic cysts. PMID- 11480737 TI - Cytologic grading of breast carcinoma. PMID- 11480738 TI - HPV testing for cervical neoplasia. PMID- 11480739 TI - Subcutaneous hyalohyphomycosis due to Fusarium oxysporum diagnosed by cytology and culture of fine needle aspirates. PMID- 11480740 TI - Tricks for obtaining better material in fine needle aspiration. PMID- 11480741 TI - Cervicofacial actinomycosis: diagnosis by fine needle aspiration cytology. PMID- 11480742 TI - Midwifing the science process. . . with confidence and humility. PMID- 11480743 TI - What is evidence-based health care? PMID- 11480744 TI - "Evidence" and midwifery practice. AB - Evidence-based care has become the new standard in the clinical disciplines. It represents a paradigm shift for clinicians, toward greater inclusion of research findings in patient care decisions. Randomized trials are the "gold standard" in clinical research and provide the strongest evidence for a treatment or intervention. But, randomized trials have limitations and cannot address all important clinical questions. Research using observational, descriptive, and qualitative methods also has a place in generating evidence for practice. Balancing the needs of individual women against what is learned from research with groups or populations is a challenge for midwives. PMID- 11480745 TI - Fostering evidence-based care in clinical teaching. AB - Understanding scientific research evidence and applying it to clinical practice is a focus in today's health care arena. Fostering evidence-based care (EBC) in clinical teaching is an effective way to help students learn to think critically, search for, evaluate, and incorporate the best research evidence into their clinical practice. For example, helping a student plan care for a woman creates a "teaching moment" for the application of scientific evidence to clinical practice. Essentially, EBC is a learned skill for both the student and the clinical teacher. It requires intellectual curiosity and a willingness to explore beyond "the way it has always been done." This article presents a review of the goals of clinical teaching, strategies to foster EBC, and tools to assess the readiness of the teacher, the student, and the clinical setting for EBC. By aligning the goals of clinical teaching with evidence-based strategies, both the educator and student strive together to provide midwifery care that is supported by available research. PMID- 11480746 TI - Evidence-based prenatal care visits: when less is more. AB - Traditionally, low-risk pregnant women in the United States who participate in prenatal care have been scheduled for approximately 14-16 prenatal visits, which is the schedule recommended by the American College of Obstetricians and Gynecologists. In 1989, an expert panel convened by the United States Department of Health and Human Services proposed a reduced frequency prenatal visit schedule for low-risk, healthy women based on the timing of specific tests or events that occur in pregnancy. Available evidence shows no adverse effect on maternal or neonatal outcomes for low-risk pregnant women who follow a reduced visit schedule, making it a highly important consideration for pregnant women and their health care providers. Other important aspects of prenatal care, especially related to adequacy and content, will be explored in-depth in a future segment of this series on evidence-based prenatal care. PMID- 11480747 TI - Evidence-based diabetes screening during pregnancy. AB - The concept of gestational diabetes was described more than a half century ago and has been studied extensively for more than 30 years. Available data indicate that the prevalence is highly variable, probably reflecting underlying risk factors. In addition, gestational diabetes is not a specific disease, but rather an abnormal laboratory value. Criteria for diagnosis are variable, and there is little agreement about who should be screened, if screening should be selective or universal, or how screening should be performed. Moreover, the most commonly used criteria in the United States differ from the European and World Health Organization standard criteria. This article describes the background for diabetes testing, current evidence for testing and diagnosis in pregnant women, "risks" of diagnosis, and various screening procedures and protocols, using data based evidence when available. Midwifery practice recommendations are also made, including examination of risk factors as clinical decisions are made about guidelines. PMID- 11480748 TI - A "conservative" approach to iron supplementation during pregnancy. AB - A review of current knowledge about iron metabolism during pregnancy and the evidence from various studies on the effects of iron supplementation in pregnancy on maternal, fetal, and infant outcomes suggest that the implicit goal of current recommendations regarding iron supplementation may be to achieve the highest hemoglobin concentration possible. This goal is only weakly related to improved maternal and infant outcomes in the current pregnancy or to improved maternal iron stores long-term. Indeed, the claim that iron supplementation is universally innocuous is shown to be controversial. For women in developed countries who are generally clinically healthy and have access to adequate nutrition, the benefits of iron supplementation are unclear, and there may be risks. Thus, a better "conservative" approach may be that such women do not require routine iron supplementation during pregnancy. The midwifery philosophy of individualizing care based on a woman's history and health status is one that should be taken in approaching the issue of iron supplementation in pregnancy. PMID- 11480749 TI - Evidence-based practice in women's health: hormone therapy for women at menopause. AB - Women's health providers, especially midwives, must take into account the known benefits and risks, as well as the "unknown," when recommending the use of hormone therapy for menopausal women, especially as it relates to heart disease, breast cancer, impaired cognition, and osteoporosis. The most recent evidence available from various studies about the benefits and risks of estrogen and hormone therapy at menopause suggests that, although hormone therapy may be protective in some women against heart disease and osteoporosis, evidence is less certain about the benefits of hormone protection against impaired cognition and the risks of breast cancer with use. The clinical approach used by midwives in which individualizing care based on each woman's health status history as well as preferences is highly appropriate for women in the perimenopausal and menopausal period. PMID- 11480750 TI - Evolution of the evidence-based Papanicolaou smear. AB - Cervical cancer rates have decreased over the past century, but rates have remained relatively constant during the last decade. Evidence-based management of the abnormal Papanicolaou (Pap) smear and the use of additional testing at the time of the Pap smear are discussed in light of the current evidence. Also explored in-depth is the evidence base for Papanicolaou smear collection and retrieval methods, especially as it relates to their influence on the validity of the smear results. In addition, issues that need to be considered by the clinician, including the use of colposcopy, especially in light of health service implications such as the need for adequately trained personnel, increased clinical visit time, and increased cost are presented. PMID- 11480751 TI - Putting risk in perspective: an evidence-based approach to selected risks associated with the use of oral contraception. AB - Although use of oral contraceptives is an effective way to prevent unintended pregnancy, concerns exist about associated adverse health outcomes. The concept of risk and different approaches to describing risk are discussed, and studies of health risks associated with specific exposures are used to illustrate the differences between relative and absolute risk. Evidence about health risks associated with use of oral contraceptives is presented within the context of population-based risk estimates. PMID- 11480752 TI - Depression in women. AB - In both developing and developed regions, depression is women's leading cause of disease burden. The burden of mental illnesses, including depression, has been seriously underestimated by traditional approaches that measure mortality and not disability. While psychiatric conditions are responsible for little more than 1% of deaths, they account for almost 11% of disease burden worldwide. PMID- 11480753 TI - Oral misoprostol and intracervical dinoprostone for cervical ripening and labor induction: a randomized comparison. PMID- 11480754 TI - Sculpting the developing brain. AB - The developing brain experiences major construction during fetal life and for at least the first decade of childhood. Many more neurons and synoptic connections are produced than are needed for later function, and the mature brain is what remains after these excess building materials are "sculpted" away. This process is thought to be the basis for the developing brain's plasticity, or the capacity to adapt its behavior and circuitry to stimulation from the external environment. Plastic reorganization of the brain is now being studied in children and adults with new noninvasive tools such as functional brain magnetic resonance imaging. This exploratory tool and other new clinical methods demonstrate how the brain's functional "maps" undergo major reorganization in response to early environmental changes. The neurobiology of brain reorganization during development is also being studied with use of new insights into the molecular mechanisms for activity dependent neuronal plasticity. Clinical disorders such as lead poisoning, metabolic and epileptic encephalopathies, and psychosocial deprivation may arise from disrupted brain plasticity. Several mental retardation syndromes and cognitive disorders recently recognized as being secondary to genetic disruption of intracellular signaling cascades may also disrupt this process. Understanding how the brain's circuitry is sculpted during development provides an important perspective for thinking about neurodevelopmental disorders. PMID- 11480755 TI - The prolonged QT syndrome. PMID- 11480756 TI - Kawasaki disease. AB - Kawasaki disease is an acute self-limited vasculitis of infancy and early hildhood that is now the leading cause of acquired heart disease in children in the United States and Japan. An infectious cause is suspected, although the etiology remains unknown. Children typically are seen with the acute onset of fever followed by signs of mucosal inflammation and vasodilatation that evolve over the first week of the illness. Laboratory tests reveal a marked systemic inflammatory response. If untreated, 1 in 4 children will develop permanent damage to the coronary arteries. Intravenous gamma globulin in conjunction with aspirin is effective in reducing the inflammation and preventing coronary artery abnormalities if administered within the first 10 days of illness. This poses a difficult situation for the pediatric care provider who must distinguish Kawasaki disease from other rash/fever syndromes in a timely manner so that appropriate therapy can be initiated. PMID- 11480757 TI - Treating obesity in youth: should dietary glycemic load be a consideration? AB - Although the adverse effects of excess adiposity on health outcomes are widely recognized, there is no consensus regarding the most appropriate dietary strategies for managing obesity in youth. Recently, a novel dietary variable termed glycemic load has been postulated to influence hunger and body weight regulation. Glycemic load, a measure of the effects of a meal on blood glucose levels, is determined by the type and the amount of carbohydrate consumed. According to a hypothetical model, ingestion of high-glycemic load meals induces a sequence of hormonal changes that alter partitioning of metabolic fuels, exacerbate hunger, and over the long-term, promote weight gain. This chapter provides an overview of the available evidence suggesting that dietary glycemic load, and its related factor, the glycemic index, should be taken into consideration in the design of weight loss interventions. PMID- 11480758 TI - Peripheral neuropathy in the first 2 years of life: diagnostic evaluation including molecular genetics. AB - Inherited polyneuropathies present in the first 2 years of life are discussed with emphasis on clinical, pathologic, and molecular data. Early-onset polyneuropathies are relatively rare, sometimes life-threatening conditions that demand early recognition by clinical and pathologic examination. Histologic and ultrastructural overlaps among the various conditions are sometimes resolved by molecular genetic analysis. The growth in disease identification by genetic localization allows a more comprehensive understanding of the clinical and morphologic heterogeneity involving rearrangements of the same gene. Molecular mechanisms explaining the acquisition of such gene rearrangements are beginning to be unraveled. Peripheral myelin disorders may be confused with primary axonal disorders, and electrophysiologic examination often helps to distinguish between these two. Furthermore, early-onset central nervous system disorders may present as peripheral polyneuropathies and confound the clinical picture. A tentative diagnosis can often be offered by pathologic examination and confirmed by biochemical enzyme analysis later. The differential clinical diagnostic considerations of early-onset polyneuropathies are offered, to help clinicians sort out these diseases in the most efficient manner. PMID- 11480759 TI - Fetal growth programs future health: causes and consequences of intrauterine growth retardation. PMID- 11480760 TI - Auditory development and hearing evaluation in children. AB - This chapter provides an overview of (1) developmental timetables relevant to hearing, and (2) current pediatric audiological techniques and practices. Structural and functional development of the auditory pathway and the development of primary auditory processing are summarized. These developmental sequences appear to follow similar paths in humans and animals. Speech and music perception involve more complex processing and are strongly influenced by experience. Hearing disorders affect the perception of complex sounds in a variety of ways, depending on the sites of lesions. Early-onset hearing impairment, including conductive loss from chronic otitis media, can seriously impede language development. Language cannot develop normally without adequate speech stimulation, and deafness is more prevalent than any other handicapping condition for which mandated neonatal screening programs exist. Sensitive and inexpensive techniques are available for performing neonatal hearing screening, and early intervention has a documented positive effect on development of language skills in hearing-impaired children. Thus, the National Institutes of Health has recommended nationwide universal neonatal hearing screening. The rationale for and the methodology of universal screening programs are summarized. Advances in the genetics of hearing impairment are reviewed. Data from these studies have influenced testing and rehabilitative protocols and have implications for future prevention and treatment of hearing impairment. PMID- 11480761 TI - Gastroesophageal reflux in infants and children. PMID- 11480762 TI - Fatal child neglect. AB - Child neglect results from either acts of omission or of commission. Fatalities from neglect account for 30% to 40% of deaths caused by child maltreatment. Deaths may occur from failure to provide the basic needs of infancy such as food or medical care. Medical care may also be withheld because of parental religious beliefs. Inadequate supervision may contribute to a child's injury or death through adverse events involving drowning, fires, and firearms. Recognizing the factors contributing to a child's death is facilitated by the action of multidisciplinary child death review teams. As with other forms of child maltreatment, prevention and early intervention strategies are needed to minimize the risk of injury and death to children. PMID- 11480763 TI - Thrombophilia in the infant and child. PMID- 11480764 TI - Iron deficiency anemia. AB - Iron deficiency anemia is the most common cause of anemia worldwide and results from inadequate iron supply for erythropoiesis. Iron deficiency is most prevalent during periods of rapid body growth: in infancy and again at puberty. Insufficient intake accounts for most cases. The clinical manifestations of iron deficiency anemia can be subtle, but irreversible delayed psychomotor development may occur if the anemia is severe and prolonged. The optimal approach is prevention and early treatment. PMID- 11480765 TI - The 22q11.2 deletion syndrome. AB - Estimates suggest that the 22q11.2 deletion occurs in approximately 1 in 4000 live births, making this disorder a significant health concern in the general population. The 22q11.2 deletion has been identified in the majority of patients with DiGeorge syndrome, velocardiofacial syndrome, and conotruncal anomaly face syndrome, suggesting that they are phenotypic variants of the same disorder. The findings associated with the 22q11.2 deletion are extensive and highly variable from patient to patient. In this chapter, we discuss the features of this disorder, with an emphasis on the clinical findings and an approach to the evaluation of these patients. In addition, we present the current understanding at the molecular level, of the genomic mechanisms and genes that are likely to play a central role in causing this frequent genetic condition. PMID- 11480766 TI - Enuresis. PMID- 11480767 TI - Advances in pediatric pharmacology, therapeutics, and toxicology. AB - This chapter reviews published studies in the field of pediatric therapeutics between July 1998 and July 2000. The most important area discussed in the first part of the chapter concerns the significant advances made in the labeling of drugs for children in the United States. Dr Harry Shirkey coined the term "therapeutic orphan" in 1968 to describe the state of children who were not being considered in either drug development or in drug clinical trials. This explains why about 80% of drugs listed in each edition of the Physicians' Desk Reference do not have labeling for the pediatric age group, especially children younger than 12 years. The recent legislative, regulatory, and pharmaceutical company activities to change this situation are summarized. These changes are current and promise to make significant contributions to the availability of drugs with adequate pediatric indications to the practicing physician. Another important change in recent years has been the appreciation of the importance of placebo controlled clinical trials for psychotropic medications in children. Trials with one of the selective serotonin reuptake inhibitors, as well as further studies involving the appropriate dosing and preparation of stimulant drugs for attention deficit/hyperactivity disorder (ADHD), are also discussed. Several new areas that promise significant knowledge in therapeutics are in the treatment of osteoporosis (a neglected condition in pediatrics), arthritis (a condition for which drugs are used to treat the disease rather than the symptoms), and acquisition of data concerning transplacental transfer of human immunodeficiency virus (HIV) and use of multiple anti-HIV drugs for treatment of this virus in the pediatric population. PMID- 11480768 TI - Wolf-Hirschhorn (4p-) syndrome. AB - Wolf-Hirschhorn syndrome (WHS) is a well-known congenital malformation syndrome caused by deletion of the short arm of chromosome 4 (4p-). In spite of more than 100 reported cases, information on its natural history remained very limited until recently. It was generally thought that these children had severe developmental disabilities and tended to be mere survivors devoid of personality. However, it is now evident that individuals with WHS are capable of more acquisition of developmental milestones than previously suggested. It is therefore very important to have guidelines for health supervision and anticipatory guidance of such patients. Although thought to affect 1 per 50,000 births, we believe that the syndrome is more common because of the many syndromes with which it is still misdiagnosed, and because only 58% of cases can be recognized on regular G-banding. The following discussion outlines the historical evolution of our recognition of the several complex aspects of this syndrome, from the very early description to the latest knowledge on clinical and cytogenetic/molecular genetic aspects. Its purpose is to draw the attention of professionals (particularly pediatricians and family practitioners) to a clinical disorder that probably affects many more individuals than previously thought. Accurate identification of such patients can lead to the organization of the most appropriate laboratory testing, to the prediction of the prognosis with relative certainty, to the development of the most appropriate health maintenance and educational plans, and to referral of the patient and the family to support groups. PMID- 11480769 TI - Factors affecting the isotopic composition of organic matter. (1) Carbon isotopic composition of terrestrial plant materials. AB - The stable isotope composition of the light elements (i.e., H, C, N, O and S) of organic samples varies significantly and, for C, is also unique and distinct from that of inorganic carbon. This is the result of (1) the isotope composition of reactants, (2) the nature of the reactions leading to formation and post formational modification of the samples, (3) the environmental conditions under which the reactions took place, and (4) the relative concentration of the reactants compared to that of the products (i.e., [products]/[reactants] ratio). This article will examine the carbon isotope composition of terrestrial plant materials and its relationship with the above factors. delta13C(PDB) values of terrestrial plants range approximately from -8 to -38%, inclusive of C3-plants ( 22 to -38%), C4-plants (-8 to -15%) and CAM-plants (-13 to -30%). Thus, the delta13C(PDB) values largely reflect the photosynthesis pathways of a plant as well as the genetics (i.e., species difference), delta13C(PDB) values of source CO2, relevant humidity, CO2/O2 ratios, wind and light intensity etc. Significant variations in these values also exist among different tissues, different portions of a tissue and different compounds. This is mainly a consequence of metabolic reactions. Animals mainly inherit the delta13C(PDB) values of the foods they consume; therefore, their delta13C(PDB) values are similar. The delta13C(PDB) values of plant materials, thus, contain information regarding the inner workings of the plants, the environmental conditions under which they grow, the delta13C(PDB) values of CO2 sources etc., and are unique. Furthermore, this uniqueness is passed on to their derivative matter, such as animals, humus etc. Hence, they are very powerful tools in many areas of research, including the ecological and environmental sciences. PMID- 11480770 TI - Physiological adaptation of crop plants to flooding stress. AB - When crop plants are subjected to soil waterlogging, or an anaerobic condition, their root and shoot systems respond differently. A variety of morphological and anatomical alterations develop in the root system. Reduction of the root respiration rate has been reported in both flooding-tolerant and intolerant species. Besides alcoholic fermentation, several diverse fermentative bypasses take place, which ameliorate the poisoning through excessive accumulation of specific metabolic intermediates. Root systems starved of oxygen are also poor providers of mineral nutrients for both themselves and the shoot systems. Stomatal closure and non-stomatal metabolic alterations are responsible for the reduction of leaf CO2 incorporation. Plant hormones are much involved in regulation of these physiological adaptations. PMID- 11480771 TI - Mechanistic studies on the inhibitory action of dietary dibenzoylmethane, a beta diketone analogue of curcumin, on 7,12-dimethylbenz[a]anthracene-induced mammary tumorigenesis. AB - Dietary factors play important roles in the carcinogenic process. The results of epidemiological data and some laboratory animal studies indicate that certain naturally occurring and synthetic components are able to block the carcinogenic process and inhibit the development of certain cancers. Dibenzoylmethane (DBM), a curcumin-related beta-diketone analogue has been reported to exhibit a remarkable inhibitory effect on 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumorigenesis in Sencar mice. The present study investigated the possible mechanisms of inhibitory action of DBM on DMBA-induced mammary tumorigenesis in mice. The summarized results indicate that: (1) in in-vitro studies. DBM inhibited DMBA metabolism and the formation of DMBA-DNA adducts in a dose dependent manner; (2) in the assay of competitive binding to estrogen receptors with [3H]-estradiol in vitro, DBM showed weak binding affinity; (3) in vivo, feeding of 1% DBM in the diet of immature Sencar mice for 4 -5 weeks decreased the uterine and parametrial fat pad weights, and lowered the serum estrogen and triglyceride levels. This study provides insight into the mechanisms involved in the inhibitory action of DBM in mouse mammary tumorigenesis. PMID- 11480772 TI - Detection of mutations in the RB1 gene by single strand conformation polymorphism (SSCP) analysis, amplification mismatch detection (AMD) analysis and polymerase chain reaction sequencing. AB - Mutations of the retinoblastoma gene are known to cause both nonhereditary and hereditary forms of retinoblastoma. Most patients with hereditary retinoblastoma have bilateral disease. Hereditary predisposition to retinoblastoma is caused by a germline mutation at the retinoblastoma gene locus (RB1) and transmitted as an autosomal dominant trait with 90% penetrance. Three quarters of these alterations represent de novo mutations. Since 75% of these cases are new mutations, there is a need for methods which can be used to identify carriers, so that informed genetic counselling will be available to patients and close relatives. In the present study, leukocyte DNA and RNA from 5 patients with sporadic bilateral retinoblastoma. were subjected to single stranded conformation analysis (SSCP) and amplification and mismatch detection (AMD) analysis. SSCP band shifts were found in 3 of the 5 patients. AMD was applied to reverse-transcriptase PCR and exons of the RB1 gene in the patients with bilateral retinoblastoma. Cleavage was found in 2 patients. Neither of these patients corresponded to the 3 with SSCP band shifts. Thus in total, 5 patients with retinoblastoma had mutations detected by a combination of SSCP and AMD analysis, and proof was sought by means of sequencing. This approach has proved to be a useful method for the rapid detection of mutations in the RB1 gene. The five mutations detected in this study were all novel and emphasise the heterogeneity of the molecular pathology in this gene. PMID- 11480773 TI - Sensitivity of mango anthracnose pathogen, Colletotrichum gloeosporioides, to the fungicide prochloraz in Taiwan. AB - In order to monitor the sensitivity of the mango anthracnose fungus, Colletotricthum gloeosporioides, to the eradicative imidazole fungicide prochloraz, a total of 43 mango orchards were surveyed throughout the Tainan area, covering a 4000 ha region of mango plantations. These orchards were recognized as having undergone higher prochloraz application. A subpopulation, 55 isolates in total, collected from Wufeng, Taichung, served as a baseline population since no fungicide was ever used in this mango plantation. A total of 545 isolates were surveyed, and it was found that the IC50s (50% inhibitory concentration) fell within a range of 0.009-0.1554 microg/ml. No significant resistance was found in the field even with higher frequency of prochloraz application. One orchard (Yujing (Wu)) located in the Yujing area known had a higher frequency of prochloraz application, the IC50s were between 0.0204 - 0.1554 microg/ml. The average IC50 was 0. 0766 microg/ml, which was about five times higher than the baseline population (0.015 microg/ml). A significant t test indicated that these two sub-populations were different at p = 0.01. The results indicated that the dose-response of C. gloeosporioides to prochloraz shifted slightly toward higher IC50 over time. A further survey using 10 microg/ml as the threshold dosage was conducted, and the results of 1375 isolates collected throughout this region showed that no isolate could survive at this dosage. Knowing that the registered dosage for field use is 83.3 microg/ml, based on above results, the author concluded that there was no sign of prochloraz resistance in mango plantations 13 years after prochloraz registration in Taiwan. PMID- 11480774 TI - Spectral characteristics of rice plants infested by brown planthoppers. AB - Spectral characteristics of rice plants at various levels of infestation by the brown planthopper, Nilaparvata lugens (Stal), (Homoptera:Delphacidae), in the early grain-filling stage were measured and analyzed using a spectroradiometer. Plant damage was classified into six scales, i.e., 0 (CK), 1, 3, 5, 7 and 9, based on the scale of infestation displayed on the surfaces of plant parts. Results showed that mean curves of reflectance spectra (350 - 1800 nm) from different scales of insect infestation were clearly differentiated, especially in the region of 737 - 925 nm, where reflectance was in the order of severity. There were significant differences in reflectance among infestations at wavelengths of 755 and 890 nm particularly. Spectral parameters such as the normalized difference vegetation index (NDVI) and cumulative reflectance may also be used to discriminate levels of infestation. Twelve wavelengths from apparent peaks and valleys of individual spectra were selected as characteristic wavelengths making up the spectral signature of each infestation. PMID- 11480775 TI - A renal hemangiosarcoma causing hematuria in a dog. AB - A 14.5-year-old male dog was presented with stranguria and hematuria of 1-month duration. Hematology and blood chemistry revealed a neutrophilia, mild azotemia and a mild decrease in the packed cell volume. Urinalysis showed high specific gravity (> 1.040 g/mL), hematuria, proteinuria and mild bilirubinuria. On physical examination, a firm oval mass located caudal to the distended urinary bladder, was palpated. Differential diagnoses included prostatitis, prostatic neoplasm, prostatic hyperplasia, and abscess. The enlarged prostate was suspected to be the cause of hematuria, and a total prostatectomy was performed. Histologically, the prostate was affected by a prostatitis with cystic papillary hyperplasia of the epithelium. The dog's condition continued to deteriorate, and death occurred 1 week later. Necropsy showed a tumor mass, approximately 5 x 4 x 3 cm in size, between the abdominal aorta and the left kidney, where the adrenal glands were embedded. Lesions were found in the kidneys, adrenal gland, lungs, heart, liver, intestine, and serosa of viscera, while the spleen was spared. This hemangiosarcoma most likely arose from the renal arteries, resulting in diffuse lesions in the kidneys thought to be the cause of hematuria. PMID- 11480776 TI - A peritoneal mesothelioma in a captive aardwolf (Proteles cristatus). AB - A 10-year-old male aardwolf (Proteles cristatus) was presented abdominal distention and emaciation for 3 months. Physical examination revealed firm abdominal masses with effusions. Cytologic assessment of the effusion showed uniform round tumor cells with a surface brush border. Necropsy showed white velvety masses covering the peritoneal surface of the liver, gall bladder, stomach, omentum, mesentery, spleen, intestine, abdominal wall and diaphragm. Histologic examination demonstrated papillary projections, lined with cuboidal tumor cells supported by fibrous connective tissue cores, arising from the serosa of visceral organs. Cytoplasmic vacuolation and a surface brush border were evident on some cells under light microscopy. Tumor cells stained positive for both cytokeratin (AE1/AE3) and vimentin. Electron microscopy showed prominent surface microvilli, rough endoplasmic reticulum, mitochondria and desmosomes in tumor cells. This may be the first reported case of peritoneal mesothelioma in a captive wild aardwolf. PMID- 11480777 TI - Professor Titus H. J. Huisman's 40 years at the Medical College of Georgia. PMID- 11480778 TI - Forty-four years (1955-1999) devoted to hemoglobin research: Titus H. J. Huisman (1923-1999). PMID- 11480779 TI - Abnormal hemoglobins: laboratory methods. AB - Laboratory methods allowing the detection and characterization of hemoglobin variants are reviewed. Protein chemistry techniques such as isoelectrofocusing, electrophoreses under various experimental conditions, cation exchange and reversed phase high performance liquid chromatography, are the most frequently used for the detection of variants. When associated with a few additional data they may lead to a presumptive diagnosis. DNA studies are also developed in many laboratories. Final identification of a variant may be achieved either by molecular biology techniques or by protein sequence analysis in which mass spectrometry now occupies a key position. PMID- 11480780 TI - Databases of human hemoglobin variants and other resources at the globin gene server. AB - Building on the pioneering efforts of Professor Huisman, several different databases of hemoglobin variants have been developed, each with progressively increased capacity for sophisticated queries and prompt updating. These resources are reviewed in the context of a larger plan for providing related resources on hemoglobins, benign and pathological variation in these proteins and the genes that encode them, and the regulation of the globin genes. PMID- 11480781 TI - Modulation of fetal hemoglobin in sickle cell anemia. AB - A partial understanding of the pathophysiology of sickle cell disease has suggested one means of treatment-increasing the distribution and concentration of fetal hemoglobin in sickle erythrocytes. Although this can be accomplished clinically with drugs like hydroxyurea, a complete understanding of the molecular and cellular basis of fetal hemoglobin regulation may suggest new and better ways of attaining this goal. PMID- 11480782 TI - The methylene tetrahydrofolate reductase (C677T) mutation as a potential risk factor for avascular necrosis in sickle cell disease. AB - Avascular necrosis (AVN) of the humeral and femoral heads is a frequent and debilitating complication of sickle cell disease. Some of the risk factors for AVN are alpha-thalassemia and age. Recently, newly discovered thrombophilia mutations have been associated with AVN in patients without sickle cell disease. We studied the frequency of the thermolabile methylene tetrahydrofolate reductase (MTHFR) variant (C677T) in adult sickle cell patients with and without AVN. The frequency of the MTHFR mutation was 35.6% in patients with AVN and 12.9% in those without AVN (p = 0.006). These data suggest that the thermolabile MTHFR variant may be a contributing risk factor for AVN in some populations with sickle cell disease. PMID- 11480783 TI - Sickle cell disease in Kuwait. PMID- 11480784 TI - Beta-thalassemia in Turkey: a review of the clinical, epidemiological, molecular, and evolutionary aspects. PMID- 11480785 TI - Genetic heterogeneity of beta-thalassemia at Cukurova in southern Turkey. AB - Beta-thalassemia is the most common genetic abnormality causing health problems worldwide. Cukurova, in the southern part of Turkey, being on the Mediterranean, is in the thalassemic belt. Since there is no cure for the disease at present, the frequency of the mutation types of beta-thalassemia must first be identified to aid in clinical follow-up and prenatal diagnosis. Carriers identified during a screening survey and patients referred to our laboratory were studied for this purpose. After routine hematological analysis molecular screening was performed by the amplification refractory mutation system and DNA sequencing. The frequency of the common mutations were: IVS-I-110 (G-->A) 57.3%, IVS-I-1 (G-->A) 8.3%, codon 39 (C-->T) 6.4%, IVS-I-6 (T-->C) 5.7%, frameshift codon 8 (-AA) 5.7%, -30 (T-->A) 4.7%, IVS-II-1 (G-->A) 3.4%, IVS-II-745 (G-->C) 2.8%, and frameshift codon 5 (-CT) 1.1%. Some rare mutations (1%) such as frameshift codon 44 (-C) 0.7%, frameshift codons 74/75 (-C) 0.7%, IVS-1-5 (G-->C) 0.7%, frameshift codons 8/9 (+G) 0.4%, frameshift codons 36/37 (-T) 0.4%, frameshift codons 22/23/24 ( AAGTTGG) 0.4%, IVS-1-130 (G-->C) 0.4%, IVS-1-5 (G-->T) 0.2%, -28 (A-->C) 0.2%, codon 15 (TGG-->TGA) 0.2%, and frameshift codons 82/83 (-G) 0.2%, were detected by sequence analysis. The codon 15 (TGG-->TGA) and frameshift codons 82/83 (-G) mutations were seen in Turkey for the first time. PMID- 11480786 TI - Hemoglobinopathies in the United Arab Emirates. AB - The use of modern DNA techniques enabled us to characterize and identify 44 distinct beta-thal mutations and nine alpha-thal genotypes in the UAE population. All of the beta-thal mutations were severe beta+ or beta0 types resulting in transfusion-dependent phenotypes. Furthermore, a large number of alphaT alleles in the alpha-thal carriers and in patients with Hb H disease, accentuate the importance of Hb H disease as a public health problem. The overall data presented here will be useful for genetic counseling, pre-marital carrier screening and the establishment of a comprehensive prenatal diagnosis program. PMID- 11480787 TI - Prenatal diagnosis of Hb H disease caused by a homozygosity for the alpha2 poly A (AATAAA-->AATAAG) mutation. PMID- 11480788 TI - Review of histological classifications of gastrointestinal epithelial neoplasia: differences in diagnosis of early carcinomas between Japanese and Western pathologists. AB - Gastrointestinal lesions considered to be high-grade adenoma/dysplasia by Western pathologists using the conventional Western classification are often diagnosed as carcinoma by Japanese pathologists using the Japanese group classification. To overcome these differences, the Padova classification, the Vienna classification, and a revision of the Vienna classification have recently been proposed. The clinical usefulness of these five classifications needs to be reviewed for early gastric, esophageal, and colorectal neoplasias. In 1998, 31 pathologists from 12 countries individually diagnosed the same 35 gastric, 21 esophageal, and 20 colorectal specimens. Their histological diagnoses can be classified conventionally and according to the newly proposed terminology, and from these data, the extent of agreement between pathologists with Western and Japanese viewpoints can be calculated, using kappa statistics. With the conventional Western, Japanese, Padova, Vienna, and revised classifications, the agreement scores were 37%, 37%, 71%, 71%, and 80%, respectively, for gastric lesions; 14%, 14%, 57%, 62%, and 67% for esophageal lesions; and 45%, 50%, 65%, 65%, and 70% for colorectal lesions. The kappa values were lower than 0.3 with the conventional Western and Japanese classifications, but higher than 0.5 for gastric lesions, higher than 0.3 for esophageal lesions, and higher than 0.4 for colorectal lesions with the newly proposed classifications. When the literature regarding treatment indications for early neoplastic lesions is reviewed, it becomes apparent that the categories of the revised classification would fit best with current clinical treatment considerations. This classification would be particularly useful for endoscopically resected specimens, to determine whether additional surgery with lymph node dissection is required. In conclusion, the use of the newly proposed terminology can, in large part, resolve the intercountry differences in the diagnosis of adenoma/dysplasia and early carcinoma. However, the newly proposed classifications should be used with caution for biopsy specimens, as sampling error may result in an underestimation of the neoplastic grade or depth of invasion. For the choice between endoscopic and surgical treatment, assessment of the depth of invasion by endoscopic inspection and ultrasound or radiography is essential. PMID- 11480789 TI - Gastric and intestinal phenotypes of gastric carcinoma with reference to expression of brain (fetal)-type glycogen phosphorylase. AB - PURPOSE: Although reports have suggested that differentiated gastric carcinomas have different phenotypes, i.e., gastric and intestinal type, this classification is complicated and can be confusing. Our previous studies have demonstrated a close relationship between carcinogenesis in differentiated-type gastric cancer and the expression of brain (fetal)-type glycogen phosphorylase (BGP). The purpose of this study was to investigate the relationship between the mucin phenotype of gastric carcinoma and BGP expression. METHODS: Ninety-six specimens of gastric carcinoma were studied using specific anti-BGP antibody. Correlation of BGP expression with intestinal and gastric phenotypes was determined with the anti-mucin antibodies, HGM, CD10, and MUC2. RESULTS: BGP was expressed in 82.6% (38/46) of differentiated type and in 24.0% (12/50) of undifferentiated type carcinomas. The incidence of BGP positivity was significantly greater in the differentiated-type carcinoma than in the undifferentiated type (P < 0.001). The proportions of gastric, mixed and intestinal types in differentiated and undifferentiated gastric carcinomas were 13.0%, 47.8%, and 39.2%, and 56.0%, 32.0%, and 12.0%, respectively. In both differentiated and undifferentiated types, the phenotype of gastric and intestinal mucin expression corresponded very well with BGP expression, that is, more than 90% of carcinomas with gastric type did not express BGP, whereas approximately 90% of carcinomas with intestinal type did express BGP. CONCLUSIONS: The classification of gastric and intestinal phenotypes of gastric carcinoma in terms of BGP expression was simpler and clearer than such classification in terms of mucin immunohistochemistry. It is suggested that BGP is a useful biomarker for the classification of intestinal and gastric type carcinoma of the human stomach, including classification from the carcinogenetic point of view. PMID- 11480790 TI - KAI1, CAR, and Smad4 expression in the progression of colorectal tumor. AB - PURPOSE: The abnormal expression of several genes is involved in the development of colorectal tumors. The aim of this study was to examine the expression of KAI1, CAR, and Smad4, which has been associated with the progression of other cancers. METHODS: We examined 12 adenomas, 38 primary carcinomas, 10 liver metastases, and 7 adenocarcinoma cell lines, by fluorescent multiplex reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: KAI1 expression was higher in adenomas (2.55 +/- 0.61; P < 0.05) and primary carcinomas (3.30 +/- 0.42; P < 0.005) than in normal mucosa, but it was not correlated with tumor stage. CAR expression was lower in Dukes' B (1.28 +/- 0.53) than in Dukes' A (2.38 +/- 0.48) (P < 0.05), but increased with tumor progression; Dukes' C (1.45 +/- 0.25), Dukes' D (1.53 +/- 0.14), and metastases (2.09 +/- 0.39) (P < 0.05). Smad4 expression increased in adenomas (2.30 +/- 0.46; P < 0.05), but decreased with tumor progression; Dukes' A (2.14 +/- 0.37), Dukes' B (1.65 +/- 0.41), Dukes' C (1.57 +/- 0.25), Dukes' D (1.08 +/- 0.18), and metastases (0.82 +/- 0.21) (P < 0.05). CONCLUSIONS: These results suggest that the upregulation of CAR and down-regulation of Smad4 are associated with the progression of colorectal tumors, while the upregulation of these genes and of KAI1 seems to be involved in the early stage. PMID- 11480791 TI - Measurement of circulating biliary glycoprotein (CD66a) in liver diseases. AB - PURPOSE: Biliary glycoprotein (BGP), a member of the carcinoembryonic antigen (CEA) gene family, is produced by hepatocytes, and is suggested to function as a cell adhesion molecule, mouse hepatitis virus receptor, and tumor suppressor. Our aim was to establish an enzyme immunoassay for circulating BGP and to study its significance in liver diseases. METHODS: For enzyme immunoassay, a monoclonal antibody (mAb), TS135, which recognizes BGP was used as a catcher, and biotin labeled polyclonal anti-CEA antibodies were used as a tracer. Seventy-six serum specimens obtained from patients with various liver diseases were submitted to the assay. RESULTS: The incidence of positivity for antigen TS135 in the serum samples of the 76 patients was 57.9%. The most significant correlation among conventional liver function tests was found between antigen TS135 and gamma glutamyl transpeptidase (gamma-GTP). However, among the 56 patients whose serum antigen TS135 and gamma-GTP levels could be measured simultaneously, 5 were antigen TS135-positive and gamma-GTP-negative (8.9%) and 6 were antigen TS135 negative and gamma-GTP-positive (10.7%). The increased serum level of antigen TS135 in 6 cholangiocellular carcinoma (CCC) patients led us to the immunohistochemical study of CCC, in which 8 of the 8 tissue specimens tested were positive for mAb TS135, indicating the production of the antigen from CCCs. CONCLUSIONS: This preliminary study suggests that the circulating antigen TS135 level correlates with gamma-GTP in liver diseases, but that TS135 may also have a unique significance, different from that of gamma-GTP, as a liver function test. PMID- 11480792 TI - Efficacy and tolerability of long-term therapy using high lamivudine doses for the treatment of chronic hepatitis B. AB - PURPOSE: A long-term follow-up study was carried out to evaluate the tolerability and efficacy of long-term therapy (1 to 3 years) with high doses (150 or 300 mg daily) of lamivudine for chronic hepatitis B. METHODS: Thirty-two patients were studied, including those who were seronegative for hepatitis B e antigen (HBeAg), as well as those with decompensated liver cirrhosis. Viral DNA clearance was monitored by using end-point dilution polymerase chain reaction (PCR), a highly sensitive method. Hepatitis B virus (HBV) polymerase gene mutations associated with resistance were determined by sequencing. RESULTS: Response to lamivudine in the sixth month was observed in 19/32 (59.4%) patients. With one exception, viral DNA results observed at this time were maintained. The YMDD mutation was detected in 12 nonresponder patients (9 YVDD, 2 YIDD, and 1 mixed population Y(V/I)DD), generally associated with the L528M mutation. Re-takeover by the wild type was observed 6 to 18 months after lamivudine withdrawal. Lamivudine response rates in noncirrhotic and cirrhotic patients were 9/18 (50%) and 10/14 (71.4%), respectively. HBeAg to anti-HBe seroconversion was found after different periods in all responder patients. Hepatitis B surface antigen (HBsAg) clearance and anti HBs seroconversion were occasionally found. CONCLUSIONS: In nonresponder patients, resistant mutants appeared up to the second year of lamivudine therapy. In spite of the presence of resistant mutants, maintenance of therapy was usually associated with a lower viral load. In responder patients, maintenance of therapy was associated with continued absence of detectable HBV DNA in serum, as monitored by highly sensitive methods. No significant side effects caused by lamivudine were observed in our patients, even in those with liver cirrhosis. PMID- 11480793 TI - Does ascorbic acid prevent retinopathy during interferon therapy in patients with chronic hepatitis C? AB - PURPOSE: Ascorbic acid was administered to patients with chronic hepatitis C to elucidate the mechanism of onset of retinopathy during interferon (IFN) therapy, and its prevention. METHODS: The subjects were 62 patients with chronic hepatitis C who had been admitted to our hospital. For the IFN therapy, 6 MIU of natural IFN-alpha, or 10 MIU of recombinant human IFN-alpha 2b was administered every day for the first 2 weeks, followed by administration three times a week for 22 weeks. The patients were randomly assigned to a group receiving 600 mg/day of ascorbic acid or a group not receiving ascorbic acid (control group). The optic fundi were examined by ophthalmologists before the IFN therapy began and subsequently at weeks 2 and 4 and then every 4 weeks during the IFN therapy. RESULTS: Retinopathy was found in 9 of the 31 patients (29%) in the ascorbic acid treated group and in 11 of the 31 patients (35%) in the control group. The cumulative incidence of hemorrhage in the ascorbic acid-treated group was lower than that in the control group during the IFN therapy, but the difference between the two groups was not significant (P = 0.186). The cumulative incidence of cotton-wool spots in the ascorbic acid-treated group was almost same as that in the control group during the IFN therapy. The median platelet counts before the therapy was begun were 11.8 x 10(4)/mm2 in the group with hemorrhage and 16.6 x 10(4)/mm2 in the group without, and the lowest platelet counts during IFN therapy were 7.3 x 10(4)/mm3 in the group with hemorrhage and 9.5 x 10(4)/mm3 in the group without, indicating significantly lower values in the group with hemorrhage (P = 0.018 and P = 0.020, respectively). The lowest platelet counts during IFN therapy were 7.4 x 10(4)/mm3 in the group with cotton-wool spots and 9.7 x 10(4)/mm3 in the group without, indicating a significantly lower value in the group with cotton-wool spots (P = 0.036). CONCLUSIONS: Ascorbic acid was not considered to be useful for the prevention of the retinopathy associated with IFN therapy in patients with chronic hepatitis C. PMID- 11480794 TI - Looping technique for transpapillary selective biopsy of the left hepatic duct. AB - PURPOSE: Because biopsy forceps tend to turn towards the right hepatic duct during endoscopic retrograde cholangiopancreatography (ERCP), selective access to the left hepatic duct is difficult. METHODS: In this study, we managed to insert biopsy forceps selectively into the left hepatic duct, by using a looping technique, in three patients. Biopsy forceps were inserted into the right hepatic duct by the conventional method. The elevator of the endoscope was kept down, and the shaft of the biopsy forceps was then advanced to the duodenal cavity until it formed a loop between the endoscope and the papilla. During the procedure, the tip of the forceps was kept at the hepatic hilus. RESULTS: In this condition, we were able to slowly rotate the tip of the forceps and direct the forceps towards the left. Sufficient material from the left hepatic duct was obtained in all patients. CONCLUSIONS: The looping technique was useful for selective access to the left hepatic duct. PMID- 11480795 TI - A second primary esophageal cancer developing 7 years after chemoradiotherapy for advanced esophageal cancer. AB - We report a rare case of advanced carcinoma and a second primary carcinoma of the esophagus, both of which were successfully cured by chemotherapy and operation at different times. In 1991, a 38-year-old Japanese man was diagnosed with advanced esophageal cancer, which was unresectable because of the bronchial invasion of the tumor. He was given chemotherapy with cisplatin (CDDP), combined with radiotherapy. During a 4-year follow-up, neither regrowth of the primary tumor nor distant metastasis occurred. In 1995, esophagoscopy demonstrated a lugol unstained region located 3 cm distal from the area of radiation to the primary lesion shown by esophagography. Histological examination of a biopsy specimen showed the mucosa to be normal. Nevertheless, yearly surveillance by endoscopy and histological examinations showed that the mucosa of the esophagus gradually began to demonstrate mild dysplasia, followed by severe dysplasia; in 1998, a diagnosis of squamous cell carcinoma was made. Esophagectomy with lymph node dissection was performed. Microscopic examination revealed that there had been pathologic complete response for the original advanced esophageal cancer. PMID- 11480796 TI - Genetic analysis of a local recurrent tumor after colonic polypectomy. AB - After polypectomy, the cut end of the polyp is usually examined by light microscopy to assess the risk of recurrent cancer. Here, we report a recurrent tumor that appeared in the colon 6 years after polypectomy, although cancer cells were not observed in hematoxylin and eosin-stained sections of the cut end of the primary polyp. Retrospectively, the primary polyp and the recurrent tumor were analyzed for mutations of the p53 gene. We detected p53 mutations in the primary polyp, even in the cut end of the polyp. The same set of two p53 mutations was detected in the recurrent tumor. These observations indicate a common origin of the primary tumor and the recurrent tumor. We conclude that it is important to analyze p53 mutations in colonic polyps, especially when the cut end of the polyp is difficult to evaluate histologically, in order to predict recurrence. PMID- 11480797 TI - Benign pseudotumorous lesion (fibroangiomyomatous hyperplasia with elastosis) in the gallbladder. AB - We describe a rare case of a benign pseudotumorous lesion (fibroangiomyomatous hyperplasia with elastosis) in the gallbladder in a 44-year-old Japanese woman, and discuss the rarity of elastosis in the gallbladder. To our knowledge, this case may be the first report of a pseudotumorous lesion of the gallbladder with elastosis in Japan. PMID- 11480798 TI - Mucoepidermoid carcinoma of the anal canal: an immunohistochemical study. AB - We present a case of mucoepidermoid carcinoma of the anal canal, with special reference to immunohistochemical analysis of the tumor to clarify its histogenesis. A 36-year-old man underwent surgery for mucoepidermoid carcinoma of the anal canal. Immunohistochemical analysis of the resected specimen was performed. Serial sections were stained immunohistochemically by the labeled streptavidin-biotin peroxidase method for various antigens, including epithelial membrane antigen (EMA); carcinoembryonic antigen (CEA); different types of cytokeratins, including CK10 and CAM 5.2; and p53 oncoprotein. The solid component of the tumor cells was immunohistochemically positive for EMA, CEA, and CAM 5.2, but negative for CK10. These staining patterns were different from those of anal squamous epithelium. These results confirm that mucoepidermoid carcinoma of the anus may arise from the anal transitional zone, and that it is biologically different from squamous cell carcinoma of the anus. PMID- 11480799 TI - Much remains to be solved in lamivudine treatment for HBV-associated liver diseases. PMID- 11480800 TI - Selecting lineup foils in eyewitness identification experiments: experimental control and real-world simulation. AB - Experimental research on eyewitness identification follows a standard principle of experimental design. Perpetrator-present and perpetrator-absent lineups are constructed with the same foils, so that the two conditions are identical except for the presence or absence ofthe trueperpetrator ofthe crime. However, this aspect of the design simulates conditions that do not correspond to those of real criminal investigations. Specifically, these conditions can create perp-absent lineups in which the foils are selected based on their similarity to an unknown person--the real perpetrator. Analysis of the similarity relations predicts that when foils for perp-absent lineups are selected based on their match to the perpetrator the false identification rate will be lower than if the foils are selected based on their match to the innocent suspect. This prediction was confirmed in an experiment that compared these two perp-absent lineup conditions. These results suggest that false identification rates in previous experiments would have been higher if the foils had been selected based on their match to the innocent suspect, rather than the absent perpetrator. PMID- 11480801 TI - The effects of limiting punitive damage awards. AB - In response to concerns that jury awards in tort cases are excessive and unpredictable, nearly every state legislature has enacted some version of tort reform that is intended to curb extravagant damage awards. One of the most important and controversial reforms involves capping (or limiting) the maximum punitive damage award. We conducted a jury analogue study to assess the impact of this reform. In particular, we examined the possibility that capping punitive awards would cause jurors to inflate their compensatory awards to satisfy their desires to punish the defendant, particularly in situations where the defendant's conduct was highly reprehensible. Relative to a condition in which punitive damages were unlimited, caps on punitive damages did not result in inflation of compensatory awards. However, jurors who had no option to award punitive damages assessed compensatory damages at a significantly higher level than did jurors who had the opportunity to do so. We discuss the policy implications of these findings. PMID- 11480802 TI - Effects of positive impression management on the Psychopathic Personality Inventory. AB - The Psychopathic Personality Inventory (PPI; S. O. Lilienfeld & B. P. Andrews, 1996) is a self-report test that has shown considerable promise as a screening measure for psychopathy. A current limitation of the PPI is that no data exist regarding the impact of response sets such as positive impression management. Although the PPI contains a validity scale (Unlikely Virtues) designed to identify response biases such as "faking good," its utility has not yet been assessed. In this study a repeated measures analogue design was employed in which 186 respondents completed the PPI both under standard conditions and with specific instructions to create a favorable impression of themselves. In the "fake good" condition, participants were able to appear significantly less psychopathic, with those who obtained higher scores in the standard instruction condition showing the largest decreases in their PPI scores. Receiver Operating Characteristic analyses indicated that, although the Unlikely Virtues scale significantly differentiated between "fake good" and honest protocols (area under the curve = .73), a considerable number of misclassifications occurred. The clinical and forensic implications of these findings are discussed. PMID- 11480803 TI - Stopping (or slowing) the revolving door: factors related to NGRI acquittees' maintenance of a conditional release. AB - The current study sought to extend the knowledge about factors associated with NGRI acquittees' maintenance of a conditional release after hospital discharge. The medical and forensic records of 125 NGRI acquittees were reviewed to collect a variety of demographic, clinical, criminal, and aftercare factors. A hierarchical survival analysis approach to determining success was compared to data analysis strategies typically employed in the area. Survival analysis, which accounts for both conditional release success status and time on conditional release, revealed that minority status, substance abuse diagnosis, and a prior criminal history were the factors that significantly predicted conditional release revocation. Treatment and policy implications of these results are discussed. PMID- 11480804 TI - Effects of past abuse experiences on children's eyewitness memory. AB - Relations between child maltreatment and children's eyewitness memory were examined. A matched sample of abused and nonabused 3- to 10-year-old children (n = 70) participated in a play session with an unfamiliar adult and were interviewed about the interaction 2 weeks later. Consistent with results from previous research, older compared to younger children's reports were more complete and accurate. Abused and nonabused children performed similarly with several exceptions: Nonabused children were more accurate in answering specific questions, made fewer errors in identifying the unfamiliar adult in a photo identification task, and (at least for younger boys) freely recalled more information. Most effects remained when group differences in IQ and behavioral symptomology were statistically controlled. Importantly, abused and nonabused children did not differ in their accuracy or suggestibility in response to questions that were relevant to abusive actions. Among abused children, however, those who suffered more severe sexual abuse made more omission errors to specific abuse-relevant questions. Contributions to psychological theory and legal implications for understanding children's eyewitness memory and testimony are discussed. PMID- 11480805 TI - Lineup administrators' expectations: their impact on eyewitness confidence. AB - This research focuses on how lineup administrators influence eyewitnesses' postidentification confidence. What happens to witness confidence when a witness makes an identification that confirms the lineup administrator's expectations; what happens when this expectation is not confirmed? In Experiment 1, participant interviewers (n = 52) administered target-absent photo lineups to participant witnesses (n = 52). The interviewers did not view the simulated crime, but were told the thief's position in the lineup. In every instance this information was false (we used a target-absent lineup). A one-way ANOVA revealed that eyewitness identification confidence was malleable as a function of interviewers' beliefs about the thief's identity. In Experiment 2, participant jurors (n = 80) viewed 40 testimonies of Experiment 1 witnesses (2 participants viewed each testimony). Participant jurors judged all participant witnesses as equally credible despite their varying levels of postidentification confidence. PMID- 11480806 TI - Clinical evaluation of packable composite resins in Class-II restorations. AB - This study investigated the clinical performance of two packable composite resins in Class-II restorations. One hundred and four Class-II restorations were placed in 52 patients by five dentists (four in dental practices and one in a university clinic) in a controlled prospective multicentre clinical trial. Each patient received one Definite/Etch & Prime 3.0 (D-EP) and one Solitaire/Solid Bond (S-SB) restoration, which were examined clinically according to modified USPHS-criteria after 1 week (baseline) and after 1 year. Statistical analysis was performed using the Wilcoxon rank sum test and the error rates method. The significance level was set to 0.05. At baseline both materials performed equally according to the evaluated criteria. After 1 year D-EP showed significantly worse marginal adaptation compared to S-SB. Both materials displayed significant deterioration for the criteria marginal adaptation, marginal discolouration, approximal contact, and fracture of restoration after 1 year compared to baseline. With a failure rate of 9.6% after 1 year, D-EP did not fulfill ADA acceptance criteria for restorative materials. It is proposed that at least 1-year data of clinical testing should be available before a new material is broadly marketed. PMID- 11480807 TI - Evaluation of pH and erosion time on demineralisation. AB - In the current study the effect of various pH values of citric acid and erosion time on erosion depth and subsurface demineralisation of human enamel was studied. In a laboratory investigation, six groups of 10 samples were eroded in 0.3% citric acid for 2 h at pH 2.54, for 2 h or 4 h at pH 3.2 and for 2 h, 4 h or 8 h at pH 4.5. The erosion depth was measured using a profilometer. The demineralised layer was then removed by ultrasonicating the samples with profilometric measurements taken at 5, 30, 120, 240 and 480 s ultrasonication time. The six test groups were classified by analysis of variance into four homogeneous groups according to their erosion depth and into three homogeneous groups according to their subsurface demineralisation. Erosion depth clearly depended on the pH value of the acid and the contact time. Depth of subsurface demineralisation and surface morphology did not depend on erosion depth. Similar subsurface demineralisation depths can be produced by appropriate combinations of pH and contact time. PMID- 11480808 TI - Oral stereognosis in stroke and Parkinson's disease: a comparison of partially dentate and edentulous individuals. AB - Oral stereognosis was measured in partially dentate and edentulous patients with stroke, Parkinson's disease, and an age and gender-matched control group. Stereognostic tests involving conventional free intra-oral manipulation of test objects were undertaken in the partially dentate and edentulous with and without complete dentures. Comparisons were made using the unpaired t-test and ANOVA. Edentulous stroke patients without dentures had significantly greater error scores and fewer correct identifications compared with partially dentate stroke patients. Stereognostic measures were similar in the partially dentate and edentulous with dentures, within experimental groups. In the partially dentate, there were no differences in stereognostic measures between the three groups. Stereognostic measures were poorer in edentulous stroke patients with and without dentures compared with the edentulous control group. Partially dentate stroke patients are less likely to have impaired oral stereognosis than edentulous stroke patients. PMID- 11480809 TI - Effect of an antibacterial varnish on lactic acid production in plaque adjacent to fixed orthodontic appliances. AB - The effect of an antibacterial varnish on lactic acid concentration in suspensions of early supragingival plaque collected adjacent to fixed orthodontic brackets was evaluated in 25 adolescents using a double-blind split-mouth design with a placebo varnish control. The test varnish contained 1% chlorhexidine and 1% thymol as active ingredients. Both varnishes were applied on two occasions within 3 days and plaque was subsequently collected at 3, 7 and 30 days after the first treatment. The samples were evaluated for total viable counts (TVC) and the proportion of mutans streptococci. Acid fermentation in suspensions was induced by glucose and L(+)-lactic acid concentrations were determined enzymatically after a 30-min incubation period. The test varnish did not affect TVC but reduced the proportion of mutans streptococci significantly at the 7-day follow-up (P<0.05). The concentration of lactic acid was reduced by approximately 20% on the 3- and 7-day follow-ups (P<0.05). The results suggest that the chlorhexidine/thymol-containing varnish may to some extent reduce the viability and metabolic activity of susceptible oral bacteria in suspensions of early supragingival plaque. PMID- 11480810 TI - Socio-economic differences in patterns of health and oral health behaviour in 25 year old Norwegians. AB - The purpose of this study was to test the hypothesis of multidimensionality of oral and general health behaviour. To evaluate the proposed behavioural dimensions, relationships with individual beliefs and socio-economic factors were explored. A simple random sample of 1190 residents born in 1972 was drawn from the populations of three counties in western Norway in February 1997. A questionnaire was mailed to the eligible sample. After one reminder, 735 subjects (58% women) replied. Principal component analysis (PCA) provided two factors, which accounted for 32.5% of the variance among the behavioural variables. One way analysis of variance revealed significant, inverse relationships between two sum scores of health enhancing and health detrimental behaviour derived from the factors; socioeconomic variables and perceived vulnerability. Controlling for gender, multiple logistic regression analyses revealed significant relationships between health enhancing behaviour and occupational status (non-manual versus student, OR=0.6, 95% CI 0.4-0.8) and perceived vulnerability (OR=2.2, 95% CI 1.5 3.0). Occupational status (manual versus student OR=1.8 95% CI 1.2-2.6 and non manual versus student OR=1.4 95% CI 1.0-2.1) turned out to be the strongest predictor of health detrimental behaviour. The present results indicate that oral and general health behaviour reflects two distinct behavioural domains. This appears to imply that oral and general health behaviour should be approached jointly in health promotion lifestyle programmes and that the lower socio economic status groups should be targeted. PMID- 11480811 TI - Application of the Third Molar Eruption Predictor to periapical radiographs. AB - The aim of this study was to test whether the Third Molar Eruption Predictor, developed for panoramic radiographs, can also be applied to periapical radiographs. The Third Molar Eruption Predictor is a transparent device (US patent 5,816,814), not commercially available, and developed for prediction of future eruption or impaction of third molars. The material consisted of periapical radiographs of mandibular third molars taken from the lateral projection without angulation of the central beam. The radiographs taken at the mean age of 20.6 years (SD +/- 1.4 years) were retrospectively available from a four-year follow-up study of third molars carried out at the University of Copenhagen, Denmark. Initially unerupted or partially erupted third molars (n=43) in 28 dental students were analyzed. Clinical statuses of the third molars were available at baseline and four years later. The device was calibrated using the method of simple proportions and Bayes' Decision Theory. The predictions made with the calibrated device were in conformity with the final clinical outcome in 84% of the cases. It was concluded that the Third Molar Eruption Predictor may also be used on periapical radiographs after calibration of the device. PMID- 11480812 TI - Cytokine concentrations in stimulated whole saliva among patients with primary Sjogren's syndrome, secondary Sjogren's syndrome, and patients with primary Sjogren's syndrome receiving varying doses of interferon for symptomatic treatment of the condition: a preliminary study. AB - Sjogren's syndrome is an autoimmune disorder which causes diminished salivary flow due to autoimmune sialoadenitis. This decrease in saliva flow is the result of inflammation and atrophy of the salivary glands. Most treatment regimens are palliative in nature, but treatment with interferon (IFN) holds promise for Sjogren's syndrome sufferers. Several studies have investigated cytokine concentrations in the salivary glandular tissues from Sjogren's syndrome patients; however, there is little information concerning cytokine expression in saliva. This is especially true with respect to treatment modalities and their effects on local cytokines. A clinical study was conducted to determine salivary interleukin (IL)-6, IFN, and IL-2, concentrations among subjects diagnosed with primary and secondary Sjogren's syndrome and a healthy control group. The primary Sjogren's syndrome showed significantly higher salivary IL-2 and salivary IL-6 than the control and secondary Sjogren's groups. There were no between group differences for salivary IFN concentrations. In addition, the study assessed salivary IL-6, IFN, and IL-2 concentrations among 18 Sjogren's syndrome patients before and after administration of IFN via the oral mucosal route. The results of the study showed that the mean values for the pre- and post-treatment groups for stimulated whole saliva flow rates were 3.15 and 3.74 ml/5 min, respectively. The post-treatment group exhibited a 16.8% increase in stimulated whole saliva flow rates. The salivary IL-6 concentration was 53.3% lower for the post-treatment group (17.79) as compared to the baseline value (33.35). The values for salivary IFN and salivary total protein were virtually unchanged from their baseline values. Salivary IL-2 values, however, were 50% lower in the post-treatment group (3.07) when compared to their respective baseline values (6.10). The results of this study suggest that healthy individuals exhibit lower salivary IL-2 and IL-6 as compared to individuals suffering from primary and secondary Sjogren's syndrome. The results also suggest that administration of IFN via the oral mucosal route may increase salivary flow rates and depress certain cytokines (IL 2, IL-6) associated with inflammatory destruction of salivary glandular tissues in Sjogren's syndrome patients. PMID- 11480813 TI - General and oral aspects of osteoporosis: a review. AB - This review comprises a short summary of up-to-date clinical knowledge on systemic osteoporosis in order to focus on the clinical dental studies on osteoporosis of the jaws, which have become available within the last 15 years. Diagnosis of jaw osteoporosis requires assessment of the bone mineral content (BMC)/density (BMD) using specially constructed jaw bone scanners and development of a corresponding gender-related set of normal BMC/BMD values for young adults, as in other sites of the skeleton. If other factors are excluded, jaw osteoporosis may involve the risk of minor accentuation of alveolar bone loss after wearing a full denture, in cases of periodontitis and in peri-implant areas. However, implant-supported overdentures conserve bone because of their positive load-related effect on the jaw. Estimation of BMC/BMD is also advisable for edentulous osteoporotic patients. Systemic treatment involving the bone metabolism affects the jaw BMC/BMD positively or negatively as in other skeletal sites, but the magnitude of this effect is site-specific. Future studies of the effect of bisphosphonates on the jaw BMC and alveolar bone loss, and analyses of the bone quality and jaw BMC/BMD in relation to implant treatment, without or with bone transplantation, are required. PMID- 11480814 TI - Gingivitis in young adults with Actinobacillus actinomycetemcomitans. AB - High intraoral load of A. actinomycetemcomitans in subjects with no or minimal periodontal disease may induce subtle changes in clinical periodontal conditions. The aim of the present study was to compare, at a site level, clinical conditions in two groups of young adults with plaque-induced gingivitis. In one group, more than 20% subgingival sites harboured cultivable A. actinomycetemcomitans (n=9), whereas in the other group, the organism was present in 20% or fewer subgingival plaque samples (n=8). Whereas no overt differences in clinical conditions could be ascertained, on average, the association between the presence of subgingival plaque and bleeding upon probing was considerably stronger (Mantel-Haenszel's common odds ratio RMH and 95% confidence interval 3.903, 2.951-5.165, P<0.001) in subjects with only a few subgingival sites harbouring A. actinomycetemcomitans as compared to subjects with a widespread intraoral distribution of the organism (R(MH)=1.637, 1.226-2.184, P<0.001). Since the proportion of sites not bleeding upon probing in the presence of supragingival plaque was slightly elevated in these subjects, the present findings may suggest a suppressed inflammatory reaction on supragingival plaque in the presence of a pronounced intraoral load of A. actinomycetemcomitans. PMID- 11480815 TI - Clinical controlled study on plaque and gingivitis reduction under long-term use of low-dose chlorhexidine solutions in a population exhibiting good oral hygiene. AB - The aim of this randomized, parallel-group, placebo-controlled, blind clinical study was to examine the dental plaque and the gingivitis inhibitory effects of two low-dose 0.06% chlorhexidine (CHX) preparations in comparison to a commercially available 0.1% CHX solution, an amine fluoride/stannous fluoride (ASF) solution and a water control as an adjunct to the daily mechanical oral hygiene measures. After prophylaxis, 81 medical students performed their tooth cleaning and were asked to rinse for 30 s, according to the instructions given by the manufacturers with the assigned amount of the following solutions: once daily 10 ml ASF 250 ppm or twice daily each 10 ml CHX 0.06% with fluoride, 10 ml CHX 0.06% without fluoride, 15 ml CHX 0.1% or 100 ml tap water. The plaque index (PlI), the gingival index (GI) and the discoloration index (DI) were taken at day -14 and months (M) 0, 1, 2, 3 and 6. In this study group with good oral hygiene, all test products led to a reduction of the PlI scores when used as supplements to the usual mechanical hygiene measures. At M3 subjects using both the 0.1% as well as the 0.06% CHX showed significantly less plaque accumulation than those using ASF. In the GI a significant difference between the control and 0.1 % CHX was found at M3 and at M6. If a further decrease in the GI is wanted, a minimum concentration of 0.1% CHX for long-term use in association with habitual cleaning is needed, which, however, should be limited to special patients. If maintaining clinical health is the goal, the other solutions are a good alternative to 0.1% CHX because they exhibit less staining. PMID- 11480816 TI - Analysis of factors affecting failure of glass cermet tunnel restorations in a multi-center study. AB - The aim of this study was to analyze factors influencing the failures of tunnel restorations performed with a glass cermet cement (Ketac Silver). Caries activity, lesion size, tunnel cavity opening size, partial or total tunnel, composite lamination or operating time showed no significant correlation to failure rate. Twelve dentists in eight clinics clinically experienced and familiar with the tunnel technique placed 374 restorations. The occlusal sections of fifty percent of the restorations were laminated with hybrid resin composite. The results of the yearly clinical and radiographic evaluations over the course of 3 years were correlated to factors that could influence the failure rate using logistic regression analysis. At the 3-year recall a cumulative number of 305 restorations were available. The cumulative replacement rate was 20%. The main reasons for replacement were marginal ridge fracture (14%) and dentin caries (3%). Another 7% of the restorations which had not been replaced were classified as failures because of untreated dentin caries. The only significant variable observed was the individual failure rate of the participating dentists varying between 9 and 50% (p=0.013). PMID- 11480817 TI - Efficacy of Australian quarantine procedures against the mosquito Aedes aegypti. AB - Methods employed by Australian quarantine officers to control Aedes mosquitoes in containers were tested against all stages of Aedes aegypti. Saltwater emersion killed all larvae but not pupae or eggs that were briefly exposed. Swimming pool chlorine, methyl bromide fumigation, and permethrin (2% active ingredient) spray provided 100% mortality of eggs, larvae, and pupae. Aerosol sprays incorporating synthetic pyrethrins are practical and also provide effective control of adults. PMID- 11480818 TI - Cost effectiveness of three arbovirus surveillance methods in northern California. AB - We compared the cost effectiveness of enzootic arbovirus surveillance in northern California by antibody detection in sentinel chickens, virus isolation from mosquitoes, and antibody detection in wild avian hosts. Total and annual recurring costs were determined for each method based on estimated personnel and actual material and travel costs for biweekly surveillance at 3 sites in the Sacramento Valley from May 1 through mid-October 1997 and 1998. Serologic detection of antibodies in wild birds was the most expensive method. Total costs associated with sentinel chickens and mosquitoes combined were less than half of those for the wild bird program. Recurring annual costs for the wild bird and mosquito methods were only slightly less than expenses for those methods during the 1st year of operation, which included nonrecurring setup costs. Recurring costs for sentinel chickens were reduced approximately 40% from total costs during the 1st year of the program and were <14% of recurring costs for wild bird serology. Exceptions and caveats of our analysis are discussed. When considering data from a companion paper on detection of enzootic virus transmission using the 3 methods, we concluded that the current system that combines sentinel chickens and virus isolation from mosquitoes is the most cost-effective and efficient surveillance program and should be retained. Future research efforts should investigate the costs and surveillance efficiency of modifications in the frequency of specimen collection and the placement of chicken flocks and mosquito traps. PMID- 11480819 TI - Laboratory evaluation of two native fishes from tropical North Queensland as biological control agents of subterranean Aedes aegypti. AB - The ability of 2 freshwater fishes, eastern rainbow fish Melanotaenia splendida splendida and fly-specked hardyhead Craterocephalus stercusmuscarum stercusmuscarum, native to North Queensland to prey on immature Aedes aegypti was evaluated under laboratory conditions. The predation efficiency of the 2 species was compared to the exotic guppy, Poecilia reticulata, which is commonly used as a biological control agent of mosquito larvae. Of the 3 fish species tested, M. s. splendida was shown to be the most promising agent for the biological control of Ae. aegypti that breed in wells. Melanotaenia s. splendida consumed significantly greater numbers of immature Ae. aegypti than P. reticulata, irrespective of developmental stage or light conditions. Unlike C s. stercusmuscarum, M. s. splendida could be handled, transported, and kept in captivity for extended periods with negligible mortality. However, M. s. splendida was also an efficient predator of Litoria caerulea tadpoles, a species of native frog found in wells during the dry season. This result may limit the usefulness of M. s. splendida as a biological control agent of well-breeding Ae. aegypti and suggests that predacious copepods, Mesocyclops spp., are more suitable. However, the use of M. s. splendida as a mosquito control agent in containers that are unlikely to support frog populations (e.g., aquaculture tanks and drinking troughs) should be given serious consideration. PMID- 11480820 TI - Chrysosporium tropicum efficacy against Anopheles stephensi larvae in the laboratory. AB - The keratinophilic soil fungus Chrysosporium tropicum was evaluated as a biological control agent against Anopheles stephensi larvae in the laboratory. Culture filtrates of C. tropicum were found to be toxic to An. stephensi larvae at various concentrations. The ranges of the 50% lethal concentration (LC50) values of fungal filtrates were 16.60-17.78, 12.02-12.88, and 34.67-35.48 microl/ml against 1st-, 2nd-, and 3rd-stage larvae, respectively. The ranges of LC99 values were 38.90-63.10, 12.02-213.80, and 74.13-109.65 microl/ml against 1st-, 2nd-, and 3rd-stage larvae, respectively. PMID- 11480821 TI - Concurrent control of mosquitoes and domestic pests by use of deltamethrin treated curtains in the New Delhi Municipal Committee, India. AB - A field trial was conducted in Block F of the Moti Bagh area of New Delhi Municipal Committee to demonstrate composite control of Anopheles stephensi and Aedes aegypti by spraying deltamethrin at 100 mg/m2 on window and door curtains of habitations. Results revealed drastic reduction (87.9-93.7%, P < 0.05) of target species in the experimental area. The impact of deltamethrin-treated curtains was also evident against nontarget species (67.9-85.7%. P < 0.05). Treated curtains provided 100% kill of An. stephensi and Ae. aegypti for 3-4 months, followed by a gradual decline in successive months. Use of deltamethrin treated curtains resulted in 92.0 reduction in slide positivity rate and 95.4% reduction in malaria cases per thousand population. The cost of deltamethrin treatment was Rs 41.15 (<$1 U.S.) per house per annum. Insecticide-treated mosquito window and door curtains, along with legislative measures, may provide cost-effective concurrent control of mosquitoes and other domestic pests. PMID- 11480822 TI - Ochlerotatus j. japonicus in Frederick County, Maryland: discovery, distribution, and vector competence for West Nile virus. AB - Ochlerotatus japonicus japonicus is reported for the 1st time south of the Mason Dixon Line, in Frederick County, Maryland. Fifty-seven oviposition trap samples were collected throughout the county between June 30 and August 24, 2000. From 971 larvae reared from the oviposition traps, 5 species were identified: Ochlerotatus triseriatus (45%), Oc. j. japonicus (43%), Aedes albopictus (7%), Culex pipiens (4%), and Toxorynchites ritulus septentrionalis (<1%). Ochlerotatus j. japonicus was found widely distributed over the area sampled. This is the 1st record of Ae. albopictus in the county as well. Vector competence studies indicated that Oc. j. japonicus is an efficient laboratory vector of West Nile (WN) virus. Depending on the viral titer at time of feeding, the estimated transmission rates for Oc. j. japonicus for WN virus were 2-4 times higher than that for Cx. pipiens. Studies of the viral titer in mosquitoes over time showed that titers in the bodies of infected Oc. j. japonicus reached their peak (approximately 10(6.5) plaque-forming units/mosquito) between 7 and 11 days after taking an infectious blood meal, and that virus became detectable in the legs (an indicator of disseminated infection) as early as 3 days after taking an infectious blood meal. PMID- 11480823 TI - Use of an infusion-baited gravid trap to collect adult Ochlerotatus japonicus. AB - Ochlerotatus japonicus adults have proven difficult to collect in significant numbers in the USA. Standard trapping and collection methods have been ineffective. Infusion-baited gravid traps, typically intended to attract Culex spp., have provided the greatest number and most consistent supply of Oc. japonicus adults collected in New Jersey. PMID- 11480824 TI - Susceptibility of adult field strains of Aedes aegypti and Aedes albopictus in Singapore to pirimiphos-methyl and permethrin. AB - The susceptibilities of field strains (Fl) of Aedes aegypti and Ae. albopictus adult females to pirimiphos-methyl and permethrin were investigated and compared with a susceptible laboratory strain (S) by using a filter paper (12 x 15 cm) impregnation method. The resistance ratio between the 50% lethal concentration values (RR50) of the Fl and the S strains shows that the RR50 of Fl Ae. aegypti for pirimiphos-methyl was 1.5 and the RR50 of Fl Ae. aegypti for permethrin was 12.9. The RR50 of Fl Ae. albopictus for pirimiphos-methyl was 1.4, and that for permethrin was 1.8. This indicates that the field strain Ae. aegypti, but not Ae. albopictus, has developed resistance to permethrin. However, both species are still susceptible to pirimiphos-methyl. Therefore, control with pirimiphos-methyl will still be effective. PMID- 11480825 TI - Efficacy of an aerosol surface spray against container-breeding Aedes. AB - The effectiveness of a residual insect surface spray against container-breeding mosquitoes was tested in Cairns, Australia. A formulation containing imiprothrin and cypermethrin (Mortein Plus Cockroach Lure 'n' Kill High Performance Surface Spray) was selected based on the label claim to "kill continuously for up to 6 months." A 1-sec spray was applied to the water and interior surface of partially flooded tires and terra-cotta pots. Treatments were paired with a control and replicated at 2 sites within 3 residential properties for a total of 6 replicates. All mosquito larvae were removed and counted weekly, and a representative sample was identified in the laboratory. Complete control of Aedes species was achieved for 4 and 5 months in all tires and pots, respectively. PMID- 11480826 TI - Ochlerotatus japonicus collected from natural rockpools in New Jersey. AB - Ochlerotatus japonicus larvae were collected from natural rockpools in New Jersey within the Delaware Water Gap National Recreational Area. Previously reported collections of Oc. japonicus in the United States have been limited to artificial containers and an occasional treehole. PMID- 11480827 TI - The use of sticky ovitraps to estimate dispersal of Aedes aegypti in northeastern Mexico. AB - A newly designed sticky ovitrap was used to determine the dispersal flight of the dengue vector Aedes aegypti in northeastern Mexico. Unfed marked females were released in the field where 100 sticky ovitraps had been positioned within a circular area 300 m in diameter. Success of this method was represented by a 7.7% (31 of 401) recapture rate during a 19-day sampling period. The maximum dispersal distance was 120 m with a mean of 30.5 m +/- 4.5 SD. Drastic hot and dry climatic conditions at the study site may have restricted mosquito dispersal. As expected, mostly gravid female mosquitoes were captured in the ovitraps. The method of sticky ovitraps proved to be useful and inexpensive for Ae. aegypti dispersal studies in Latin American countries. In addition to the marked mosquito populations, unmarked Ae. aegypti, Culex sp., roaches, and even small vertebrates such as lizards and sparrows were found glued to the sticky ovitraps. Based on these studies, the use of sticky ovitraps is recommended for Ae. aegypti dispersal studies, specially for dengue control programs in developing countries. PMID- 11480828 TI - Evidence for a new sibling species of Anopheles minimus from the Ryukyu Archipelago, Japan. AB - The Anopheles minimus complex is known to comprise at least 2 sibling species (A and C) in Thailand and Vietnam. This study investigated the specific status of An. minimus on Ishigaki Island, the Ryukyu Archipelago, Japan using morphological and genetic analyses. Morphological studies revealed that almost all (99.5%) of the adult mosquitoes are characterized by the humeral pale spot on the costa of their wings, a character that partially differentiates species A and C elsewhere. A high frequency (81.4%) have a pale fringe spot at the tip of vein 1A, a character rarely observed in other An. minimus populations. Significant seasonal variation in the size of wild An. minimus mosquitoes on the island was observed, with the largest size in the winter. Scanning micrographs of the cibarial armature of females from Ishigaki Island revealed that over 90% had cone filaments clearly differing in shape from those of species A or C. The Giemsa stained metaphase karyotypes of larval brain cells were somewhat similar to those of species A, with a few exceptions, but were very different from those reported for species C. Crossing experiments between species A (CM strain) from Thailand and the progeny of An. minimus from Ishigaki Island (ISG strain) revealed postzygotic genetic incompatibility, although no prezygotic isolation. Hybrid progeny were only obtained from CM female x ISG male. F2 hybrid progeny were not obtained, since the hybrid males were sterile or almost sterile with atrophied testes or abnormal spermatozoa, although the polytene chromosomes of hybrid larvae showed synapsis. The hybrid females backcrossed with either CM or ISG males laid eggs with significantly lowered fertility and viability. The sequence for the D3 region of the 28S gene of ribosomal DNA of the ISG strain differed from those of species A and C. In addition, sequence data from Vietnamese mosquitoes suggest that the An. minimus complex may contain additional species. The morphological, cytogenetic, molecular, and hybridization evidence together suggest the existence of another sibling species of the An. minimus complex on Ishigaki Island, which is provisionally designated An. minimus species E. PMID- 11480829 TI - Bulimics' responses to food cravings: is binge-eating a product of hunger or emotional state? AB - This study examined the roles of hunger, food craving and mood in the binge eating episodes of bulimic patients, and identified the critical factors involved in the processes surrounding binge-eating episodes that follow cravings. This was a prospective study of the binge-eating behaviour of 15 women with bulimia nervosa. The participants used food intake diaries and Craving Records to self monitor their nutritional behaviour, hunger levels and affective state. Cravings leading to a binge were associated with higher tension, lower mood and lower hunger than those cravings not leading to a binge. Levels of tension and hunger were the critical discriminating variables. The findings of the study support empirical evidence and models of emotional blocking in binge-eating behaviour and challenge the current cognitive starve-binge models of bulimia. The role of food cravings in the emotional blocking model is discussed in terms of a classically conditioned motivational state. Implications for treatment are addressed. PMID- 11480830 TI - On fatalism, pessimism, and depressive symptoms among Mexican-American and other adolescents attending an obstetrics-gynecology clinic. AB - There is growing evidence that Mexican-American adolescents may be at increased risk for depressive symptoms. We sought to replicate and explain this result in a study of adolescent girls attending an obstetric-gynecologic clinic. Three hundred and four girls of diverse ethnic backgrounds completed measures of depressive symptoms, negative attributional style, and locus of control. Consistent with predictions, we found that Mexican-American adolescent girls reported more depressive symptoms than adolescent girls from other ethnic backgrounds, and that Mexican-American adolescent girls displayed more negative cognitive styles than girls from other ethnic backgrounds. Depression differences appeared to be partly explained by differences in negative cognitive style. Implications of the results for a theory of increased Mexican-American adolescent depression, and for applied work, were discussed. PMID- 11480831 TI - Danger Ideation Reduction Therapy (DIRT) for treatment-resistant compulsive washing. AB - Five intractable cases of obsessive-compulsive disorder were treated with the Danger Ideation Reduction Therapy (DIRT) program. All five cases: (1) had displayed excessive washing/cleaning behaviour for at least 10 years; (2) had failed to respond to a minimum of two separate, 12-week drug trials with serotonergic agents; (3) failed to respond to at least 15 sessions of exposure and response prevention at the beginning of the present trial, and; (4) satisfied the DSM-IV criteria for OCD with Poor Insight. DIRT was conducted in 14, weekly, individual sessions or until, in the judgement of the treating clinician: (1) clinically significant gains were apparent with minimal symptomatology remaining, and (2) clients displayed a sound grasp of the cognitive model underpinning DIRT procedures. At post-treatment, substantial reductions in scores on the Padua Inventory (PI), Maudsley Obsessional-Compulsive Inventory (MOCI), Beck Depression Inventory-II (BDI-II) and two global rating scales were apparent for four of the five subjects. These improvements were maintained at 4-to-6 month follow-up, with four of the five cases meeting Jacobson and Truax's (1991) criteria for 'recovery' on the MOCI and the PI by this assessment stage. While one subject remained non-responsive, the present findings suggest that DIRT may be a viable option for treatment-resistant cases of compulsive washing. The theoretical implications of the findings are discussed. PMID- 11480833 TI - Defensiveness predicts peer rejection of depressed children. AB - The present study examined the ironic possibility that defensiveness, far from suppressing depression and staving off negative interpersonal consequences, is actually associated with interpersonal difficulties (e.g. peer rejection). Participants were 72 youth psychiatric inpatients (aged 7-17 years, mean+/-SD 13.18+/-2.59 years), who completed self-report measures of defensiveness and depression. Chart diagnoses were available, and peer rejection ratings were collected. Results indicated that depressed children with a defensive style obtained the highest peer rejection ratings, other children, including depressed but non-defensive children, were not as negatively rated by peers. Implications regarding self-presentation and depression, as well as regarding clinical work with depressed people, were discussed. PMID- 11480832 TI - Memory and memory confidence in obsessive-compulsive disorder. AB - Pathological doubt, often found in individuals with obsessive-compulsive disorder (OCD), has been theoretically linked to memory deficits, but empirical evidence for such deficits has been mixed. In contrast, many studies suggest that individuals with OCD have low confidence in their memories. The present study aimed to build upon previous research by measuring memory accuracy and confidence in OCD using ecologically valid, idiographically-selected stimuli. Individuals with OCD (OCs), anxious controls (ACs), and nonanxious controls (NACs) were exposed to a set of objects that the OCs had identified as safe, unsafe, or neutral. Participants were then asked to recall as many objects as possible and to rate their confidence in each memory. This process was repeated 6 times, using the same stimuli for each trial. Contrary to hypothesis, no group differences emerged in memory accuracy. However, OCs' memory confidence for unsafe objects showed a progressive decline over repeated trials. This pattern was not observed among NACs or ACs. Furthermore, OCs with primary checking reported lower confidence in long-term memory than did OCs without primary checking. These results suggest that when OCs are repeatedly exposed to threat-related stimuli (such as repeated checking), their level of confidence in remembering these stimuli paradoxically decreases. PMID- 11480834 TI - Cognitive-behavior therapy for discontinuation of SSRI treatment of panic disorder: a case series. AB - In this report we describe the outcome of eight outpatients with panic disorder and agoraphobia who discontinued their treatment with a selective serotonin reuptake inhibitor (SSRI) in the context of a structured, group program of cognitive-behavior therapy. All patients successfully discontinued their SSRI medication while demonstrating clinical improvement. These results were maintained at 3-month follow-up. This case series suggests that manualized CBT for discontinuation of benzodiazepine treatment for panic disorder may be successfully applied to SSRI discontinuation as well. PMID- 11480835 TI - Distinguishing obsessive features and worries: the role of thought-action fusion. AB - Obsessions are a key feature of obsessive-compulsive disorder (OCD), and chronic worry is the cardinal feature of generalized anxiety disorder (GAD). However, these two cognitive processes are conceptually very similar, and there is a need to determine how they differ. Recent studies have attempted to identify cognitive processes that may be differentially related to obsessive features and worry. In the current study we proposed that (1) obsessive features and worry could be differentiated and that (2) a measure of the cognitive process thought-action fusion would distinguish between obsessive features and worry, being strongly related to obsessive features after controlling for the effects of worry. These hypotheses were supported in a sample of 173 undergraduate students. Thought action fusion may be a valuable construct in differentiating between obsessive features and worry. PMID- 11480836 TI - Panic symptoms during trauma and acute stress disorder. AB - This study investigated the role of panic symptoms that occur during trauma and subsequent acute stress disorder (ASD). Civilian trauma (N=51) survivors with either acute stress disorder (ASD), subclinical ASD, or no acute stress disorder (non-ASD) were administered the Acute Stress Disorder Scale, Impact of Event Scale, Beck Anxiety Inventory, Beck Depression Inventory, and the Anxiety Sensitivity Index (ASI). Participants also completed the Physical Reactions Scale to index panic symptoms that occurred during their trauma. Overall, 53% of participants reported panic attacks during their trauma. ASD and subclinical ASD participants reported more peritraumatic panic symptoms, and higher ASI scores, than non-ASD participants. These findings are consistent with the notion that peritraumatic panic may be related to subsequent posttraumatic stress, and suggest that modification of maladaptive beliefs about physical sensations should be addressed in posttraumatic therapy. PMID- 11480837 TI - Memory for facial expressions in social phobia. AB - Memory biases toward threat have been documented in several anxiety disorders, but contradictory findings have recently been reported in social phobics' recognition of facial expressions. The present study examined recognition memory in clients with social phobia, in an effort to clarify previous inconsistent results. Just before giving a speech to a live audience, social phobia clients and normal controls viewed photographs of people with reassuring and threatening facial expressions. The stimuli were later presented again alongside photographs of the same person with a different facial expression, and participants chose which face they had seen before. Individuals with social phobia were less accurate at recognizing previously seen photographs than controls, apparently due to state anxiety. In contrast, social phobics did not show a memory bias toward threatening facial expressions. Theoretical and treatment implications are discussed. PMID- 11480838 TI - The validity of the PTSD checklist as a measure of symptomatic change in combat related PTSD. AB - Little research to date has examined the ability of self-report measures to assess changes in symptom severity and diagnostic status as a function of treatment. This study investigated the validity of the posttraumatic stress disorder (PTSD) checklist (PCL) as a measure of symptomatic change following programmatic treatment. A sample of 97 Vietnam veterans with combat-related PTSD was assessed using the clinician-administered PTSD scale (CAPS) and the PCL prior to, and 9 months following, participation in a PTSD treatment program. Using the CAPS as the "gold standard" measure of PTSD symptomatology, the PCL demonstrated high diagnostic accuracy pre- and posttreatment. However, significant variations in accuracy were evident in the ability of the PCL to determine the presence and severity of individual symptoms at each time point. In addition, as symptoms improved from pre- to posttreatment, and approached the threshold criteria, the PCL demonstrated reductions in diagnostic accuracy. As a measure of overall symptomatic change, the PCL underrated improvement in comparison to the CAPS. The results supported the use of an overall cut-off score of 50 on the PCL for a diagnosis, and an item score of 3 for symptom criterion, in this population. PMID- 11480840 TI - Alternative therapies in rheumatic diseases--pros and cons. PMID- 11480839 TI - Development and initial validation of the obsessive beliefs questionnaire and the interpretation of intrusions inventory. AB - In 1995 the Obsessive Compulsive Cognitions Working Group initiated a collective process to develop two measures of cognition relevant to current cognitive behavioural models of OCD. An earlier report (Behav. Res. Therapy, 35 (1997) 667) describes the original process of defining relevant domains. This article describes the subsequent steps of the development and validation process: item generation, scale reduction, and initial examination of reliability and validity. Two scales were developed. The Obsessive Beliefs Questionnaire consists of 87 items representing dysfunctional assumptions covering six domains: overestimation of threat, tolerance of uncertainty, importance of thoughts, control of thoughts, responsibility, and perfectionism. The Interpretation of Intrusions Inventory consists of 31 items that refer to interpretations of intrusions that have occurred recently. Three of the above domains are represented: importance of thoughts, control of thoughts, and responsibility. The item reduction and validation analyses were conducted on clinical and non-clinical samples from multiple sites. Initial examination of reliability and validity indicates excellent internal consistency and stability and encouraging evidence of validity. However, high correlations indicating overlap between some of the scales, particularly importance of thoughts, control of thoughts, and responsibility will need to be addressed in subsequent empirical and theoretical investigations. PMID- 11480841 TI - International classification criteria for antiphospholipid syndrome: synopsis of a post-conference workshop held at the Ninth International (Tours) aPL Symposium. AB - An international workshop on classification criteria for antiphospholipid syndrome (APS) was held on 16 September 2000 in Tours, France, following the Ninth International APS Symposium. The workshop addressed issues that were not resolved by the previous (1998) international workshop on the classification criteria. Participants at the workshop agreed that no changes should be made to the international (Sapporo) criteria at the present time. However, to improve the criteria, future efforts should be focused on the following: (1) further evaluation of the international (Sapporo) criteria for definite APS; (2) definition of other categories of APS such as 'probable' and 'possible' APS; (3) guidelines for the clinical diagnosis as distinct from classification of APS; (4) strategies to improve the compliance of laboratories worldwide, with recommended procedures for Lupus anticoagulant (LA) and anticardiolipin (aCL) assays; (5) development of monoclonal antibody standard reagents for aCL and LA assays; and (6) refinement and subsequent evaluation of antibeta2GPI assays for use in idenfitication of APS. PMID- 11480842 TI - Prevention of autoantibody formation and prolonged survival in New Zealand Black/New Zealand White F1 mice with an ancient Chinese herb, Ganoderma tsugae. AB - For centuries, Chinese medicine has regarded Ganoderma, a fungus (Myceteae, Amastigomycota, Busidomycetes, Aphyllophorales, Polyporaceae, Ganoderma) also known as 'Ling Zhi' in Mandarin, as a premium remedy for many diseases. Until now, no convincing data regarding its therapeutic effects in vivo on autoimmune diseases have been demonstrated. In this study, a controlled protocol was conducted in which New Zealand Black/White F1 mice were fed standard chow with prednisolone (0.5 mg/kg/day) or Ganoderma tsugae extract, commencing at 2 months of age. It was found that the F1 mice responded well to Ling Zhi extract. Ling Zhi improved the survival rate of lupus mice, decreased the amount of proteinuria, decreased serum levels of anti-dsDNA autoantibody, and showed evidence of decreased perivascular and parenchyma mononuclear cell infiltration in vital organs. PMID- 11480843 TI - FcgammaRIIa/IIIa polymorphism and its association with clinical manifestations in Korean lupus patients. AB - The aim of this study was to determine the distribution of the FcgammaRlla and FcgammaRIIIa polymorphisms and their association with clinical manifestations in Korean lupus patients. Three hundred SLE (systemic lupus erythematosus) patients (48 male, 252 female) meeting 1982 ACR criteria and 197 Korean disease-free controls were enrolled. Genotyping for FcgammaRlla 131 R/H and FcgammaRIIIa 176 F/V was performed by PCR of genomic DNA using allele-specific primers and the FcgammaRIIIa genotype was confirmed by direct sequencing of PCR product in some cases. There was significant skewing in the distribution of the three FcgammaRIIa genotypes between the SLE and the controls (P=0.002 for R/R131 vs R/H131 and H/H131, OR 2.5 (95% Cl 1.4-4.5), but not in FcgammaRIIIa genotypes. FcgammaRIIa-R allele was a significant predictor of lupus nephritis, as compared with SLE patients without nephritis (P=0.034 for R131 vs H131, OR 1.4 (95% Cl 1.03-1.9)), but proliferative nephritis (WHO class III and IV) was less common in patients with FcgammaRlla-R/R131 and in FcgammaRIIa-R allele. In 300 SLE patients, high binding allele combination H131/V176 was less common in SLE with nephritis than in SLE without nephritis. Hemolytic anemia was less common in R131/F176 allele combination among four FcgammaRIIa/FcgammaRIIIa allelic combinations. Male SLE patients showed a higher frequency of renal involvement, serositis, thrombocytopenia, malar rash and discoid rash than female SLE, and male SLE had a higher frequency of FcgammaRIIa-R/R131 or R131-allele than male controls, but FcgammaRIIa or FcgammaRIIIa genotypes had no association with renal involvement in male SLE patients. FcgammaRIIa-H/H131 showed a higher frequency of hemolytic anemia and less pulmonary complications in male SLE. Female SLE patients showed higher frequency of any hematologic abnormality, lymphopenia, anticardiolipin antibody (+) and anti-Ro antibody (+) than male SLE, and had earlier onset of first symptoms. There was no skewing in FcgammaRIIa or FcgammaRIIIa genotypes between female SLE and female controls, but FcgammaRIIa-R131 allele showed skewing between female SLE with nephritis and female SLE without nephritis. The age at onset of thrombocytopenia was earlier in FcgammaRIIa R/R131 among three FcgammaRIIa genotypes, and serositis in FcgammaRIIIa-F/F176 among three FcgammaRIIIa genotypes. FcgammaRIIa-R131 homozygote was a major predisposing factor to the development of SLE and FcgammaRIIa-RI31 homozygote and R131 allele were a predisposing factor, and H131/V176 was a protective allele combination in lupus nephritis. In contrast to other ethnic patients, in our study cohort, clinical manifestation was different between male and female, and FcgammaRIIa and FcgammaRIIIa showed somewhat different clinical associations between the genders. PMID- 11480844 TI - Polymorphic light eruption and the HLA DRB1*0301 extended haplotype are independent risk factors for cutaneous lupus erythematosus. AB - Recent evidence suggests that polymorphic light eruption (PLE) is an inherited photosensitivity disorder which may predispose to cutaneous lupus erythematosus (LE). In this study we examine the relative risk (RR) attributable to the presence of PLE, together with the effect of the major histocompatibility complex (MHC) in the development of cutaneous LE. Eighty-five Caucasian patients with annular subacute cutaneous LE (SCLE) and discoid LE (DLE) were recruited, together with 102 first degree relatives and 200 healthy local Caucasian controls. Symptoms suggestive of PLE were elicited in patients and relatives, and human leukocyte antigen (HLA) typing determined by PCR-SSP. Standard association analysis and family transmission disequilibrium testing (TDT) were then used to compare the HLA frequencies between groups. We found a significant (P < 0.05) association of the HL4 A*01, B*08, DRB1*0301 extended haplotype with both SCLE and DLE and also significant association of DLE with the HLA A*03, B*07, DRB1*15 haplotype, with a possible protective effect in SCLE for HLA B*44 and DRB1*04 (P=0.002 and 0.001 respectively). Association was observed between PLE and cutaneous LE (P < 0.001), but not between PLE and any HLA allele. From these figures we estimate, for the general population, that the RR of developing SCLE given the presence of (a) PLE, (b) DRB1*0301 and (c) both PLE and DRB1*0301 is 3.37, 5.45 and 12.03, respectively. For DLE, equivalent RRs are 3.11, 2.15 and 6.94. In conclusion, these data imply the involvement of both PLE and HLA DRB1*0301 in the development of SCLE and DLE. They form a basis for examining the genetic architecture of photosensitivity, some aspects of which may be common to both cutaneous LE and PLE. PMID- 11480845 TI - Benefits of leflunomide in systemic lupus erythematosus: a pilot observational study. AB - The objective of this study was to retrospectively explore the safety and efficacy of leflunomide (LEF) in outpatients with systemic lupus erythematosus (SLE). Eighteen SLE females received LEF, open label, 100 mg/day loading dose for 3 days followed by 20 mg per day. Patients were evaluated for safety and efficacy after 2-3 months of therapy. The mean age was 42.6 y and mean disease duration 7.9 y. ACR criteria were met by 15/18. Four patients stopped LEF during the observation period. Ten of 14 LEF-treated patients had subjective improvement with 9/14 patients achieving lower SLEDAI scores. The mean SLEDAI decreased by 2.1 (P=0.005) and the mean ESR decreased by 9mm/h (P=0.02). Prednisone dosages could be reduced in 2/5 subjects without a flare. No organ-threatening or life threatening side effects were seen in our patients. Diarrhea occurred in seven patients (two stopped LEF), rash occurred in one patient (stopped LEF), one patient stopped LEF for reasons not related to therapy. Blood pressure was unchanged. Leflunomide was efficacious and safe in this cohort of SLE patients after 2-3 months of therapy. Placebo-controlled trials of longer duration are indicated. PMID- 11480846 TI - Presence of cutaneous interferon-alpha producing cells in patients with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) patients have increased levels of interferon alfa (IFN-alpha) in the circulation but a reduced number of functionally intact natural IFN-alpha producing cells (IPC) in peripheral blood. In search for tissue localisation of activated IPC, we investigated skin biopsies from SLE patients for the occurrence of such cells. Eleven SLE patients with inflammatory skin lesions and six healthy controls were biopsied. An immunohistochemical technique (IH) and in situ hybridisation (ISH) were used to detect intracellular IFN-alpha protein and IFN-alpha mRNA, respectively. In all 11 biopsies from SLE lesions, a high number of IPC were detected by IH. In the nonlesional SLE biopsies we could also demonstrate IPC in 10/11 patients. In 6/11 SLE patients, IFN-alpha mRNA containing cells could be detected in the specimens. A low number of IPC were detected in 1/6 healthy controls by IH, but no ISH positive cells were seen. Our results demonstrate that SLE patients have active IPC in both dermal lesions and in noninflammatory skin. A recruitment of IPC from blood to peripheral tissues may explain the low number of circulating natural IPC in SLE patients. Because the type I IFN system is involved in the SLE disease process, these results are of interest for the understanding of the pathogenesis in SLE. PMID- 11480847 TI - Subacute abdominal pain requiring hospitalization in a systemic lupus erythematosus patient: a retrospective analysis and review of the literature. AB - In the systemic lupus erythematosus (SLE) patient, abdominal pain is a common problem. Intraabdominal vasculitis must be excluded as the source because of its potentially high mortality rate. We retrospectively reviewed the charts of 56 SLE patients with 75 admissions for predominantly subacute abdominal pain (abdominal pain without peritoneal signs) severe enough to require hospital admission, comparing the diagnostic modalities used, ultimate diagnoses, and use of corticosteroids before admission with 56 age- and sex-matched patients without SLE admitted for abdominal pain during the same time interval. SLE patients were further subdivided by disease activity at presentation using the SELENA SLEDAI score. The in-hospital mortality for all patients in this review was 0%. There were no statistically significant differences in the use of computed tomography between SLE and control patients. Intestinal vasculitis was diagnosed in 5.4% of SLE patients compared with 0% of control patients (P = 0.0433). Only patients with SLEDAI scores >8 developed vasculitis (P < 0.001). We recommend the routine use of computed tomography to diagnose vasculitis only in patients with SLEDAI scores >8 and subacute abdominal pain. All SLE patients with SLEDAI scores <8 and subacute abdominal pain should be evaluated for a cause of abdominal pain other than vasculitis. PMID- 11480848 TI - Hyperactivity in a mouse model of the antiphospholipid syndrome. AB - In the antiphospholipid syndrome (APS), antibodies to a complex of phospholipids and beta2-glycoprotein I (beta2-GPI) are associated with recurrent thromboembolic events, spontaneous abortions, thrombocytopenia and central nervous system (CNS) disturbances. Animals immunized with beta2-GPI develop the systemic manifestations of APS but the involvement of the (CNS) in these animals has not been studied. The objective of the present study was to examine mice with induced experimental APS for behavioral changes. Female Balb/C mice were immunized once with beta2-GPI in complete Freund's adjuvant (CFA) or with CFA alone. Four months after immunization the mice were tested in the staircase apparatus and the following two variables were measured: (1) number of rears: and (2) number of stairs climbed by the mice. Immunization with beta2-GPI resulted in elevated levels of circulating anti-negatively charged phospholipids and anti-beta2-GPI antibodies. The APS mice exhibited hyperactive behavior as reflected by more frequent rears (P < 0.023) and higher number of stairs climbed (P < 0.019) by the mice in 3 min. This simple test demonstrated that experimental APS animals are significantly hyperactive and may serve as a marker for CNS involvement in this model. PMID- 11480849 TI - Malignancies during follow-up in an epidemiologically defined systemic lupus erythematosus inception cohort in southern Sweden. AB - The objective of this study was to identify all malignancies in an inception cohort of SLE patients in southern Sweden and compare with the observed frequencies and spectrum of malignancies in the general population. All adult incidence cases of SLE in a defined population during the period 1981-1996 were retrieved from a prospective database and the cases were followed to endpoint or through 1998. The SLE cohort registry was aggregated with the National Cancer Registry to identify all malignancies by date, type and outcome. Standardized morbidity rates (SMR) were calculated based on the sex- and age-matched general population of the region. Sixteen malignancies occurred in 13 patients out of a total of 116 SLE patients observed for 1086 patient-years. The SMR for all cancers detected was 2.24 (confidence interval 0.6-5.7) for males and 1.02 (confidence interval 0.4-2.1) for females and thus indicative of no general increase in malignancies. However, the SMR for non-Hodgkin lymphoma was 11.63 (confidence interval 1.4-42.0), for pulmonary cancer 5.55 (confidence interval 0.7-20.1) and prostatic cancer 6.41 (confidence interval 1.3-18.7) all significantly increased. The increase in prostatic carcinoma disappeared when only cases occurring after a latency period of 3y after SLE diagnosis were included. In this comprehensive inception cohort of SLE no increase in relative risk of malignancy overall was found, but the frequencies of non-Hodgkin lymphoma and pulmonary cancer were increased, possibly also the frequency of prostatic carcinoma. PMID- 11480850 TI - Infections in outpatients with systemic lupus erythematosus: a prospective study. AB - The objective of this study was to assess the incidence and risk factors of infections in 200 SLE outpatients. All outpatients with active or inactive SLE without infections in the previous month were included. They were assessed every 3 months. Major infections were those requiring hospitalization and parental antibiotic therapy; minor infections required oral or topical therapy. Sociodemographic, disease activity using the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), therapy and laboratory variables were evaluated. After a follow-up of 22+/-7 months, 65 (32%) patients had infections; 35% of those were major. The most common sites for infection were urinary (26%), skin (23%), systemic (12%), and vaginal (9%). At infection onset, 50 of 65 patients (77%) had disease activity, with a mean SLEDAI score of 6.1. The variables significantly associated with infection in the univariate analyses were the presence of disease activity, SLEDAI score, renal activity, prednisone dose, and IV cyclophosphamide. The only variable associated with infection in the multivariate analyses was a SLEDAI score of 4 or higher. Most infections in SLE outpatients were single, minor, non-life threatening, and associated with disease activity independently of sociodemographic and therapeutic factors. PMID- 11480851 TI - Two- and three-dimensional echocardiography in primary antiphospholipid syndrome: misdiagnosis as rheumatic valve disease. AB - This is a report of a woman in the fifth decade of life with primary antiphospholipid syndrome and involvement of a heart valve. Diagnosis was reached with echocardiography and serological studies. PMID- 11480852 TI - Lupus profundus: not a benign disease. AB - Lupus profundus is an unusual clinical variant of cutaneous lupus erythematosus that has previously been described as a benign disease that follows a mild course. This report describes the extent of disease and associated comorbidities in patients with severe lupus profundus and systemic lupus erythematosus. Four cases of lupus profundus are reviewed and their associated systemic disease complications are highlighted. All four patients fulfilled at least four of the 11 criteria for systemic lupus erythematosus. One patient suffered from severe facial disfigurement and Parry-Romberg syndrome. Two patients developed nonhealing ulcers on the scalp. All four patients had scarring alopecia as well as depressed areas over large areas of their body surfaces. All patients were resistant to conservative therapy, and required long-term aggressive therapy. Clinical depression secondary to disfigurement was a major problem in three patients. Extensive lupus profundus may be associated with more serious systemic disease and warrants aggressive treatment early on to prevent permanent disfigurement and its resultant psychological consequences. PMID- 11480853 TI - Quantitative comparison of human endogenous retrovirus mRNA between SLE and rheumatoid arthritis. PMID- 11480854 TI - Cerebral blood flow in systemic lupus erythematosus. PMID- 11480855 TI - Technetium generator log form. AB - Technetium generators are loaded with molybdenum-99 (99Mo), that has a 66-h half life and decays to technetium-99 m (9mTc). 99Tc with a single 140 keV gamma photon emission and a 6-h half life is an ideal isotope for nuclear medicine imaging. Many nuclear medicine units receive generators and use the 99Tc elution to label a variety of prepared chemical species for the majority of the studies they perform. Other options are receiving bulk 99mTc and doing the same tagging or getting unit doses of already labeled pharmaceuticals. Units using generators must meet requirements spelled out in regulations of the Nuclear Regulatory Commission, Title 10, CFR, parts 20, 35, and 71 as well as the Department of Transportation, Title 49, CFR, part 173. The form described in this paper was an attempt to get all of the required records for each individual generator on a single sheet of paper. PMID- 11480856 TI - University of Rochester laboratory inspection checklist. AB - In early 1999, the University of Rochester significantly revised our laboratory inspection program. As part of this revision, we developed a new checklist to be used during laboratory inspections to help promote consistency between inspections and between inspectors. PMID- 11480857 TI - Computer modeling codes for radiological and chemical airborne releases. AB - Brief descriptions are provided for commonly used computer codes for estimating the consequences of atmospheric releases of radionuclides and chemicals. Different levels of sophistication and input detail are necessary for emergency response versus emergency planning. PMID- 11480858 TI - Radiation safety considerations for large, multi-user laboratories. AB - Many research institutions are adding new laboratory space in the form of large, multi-user rooms with an open architecture. Unfortunately, many of the features that make such rooms attractive to researchers make them difficult to manage from a radiation safety perspective. In particular, coping with radiological incidents and enforcing radiation safety standards are difficult when several research teams share a common room. This paper discusses the problems noted in such laboratories at the University of Rochester and Yale University and describes the manner in which some of these problems have been addressed. PMID- 11480859 TI - Determining the solubility of aqueous radioactive waste from a pharmaceutical research and development lab. AB - The management of radioactive waste disposal at pharmaceutical research facilities is often the responsibility of the Radiation Safety Office. Aqueous waste containing a variety of radionuclides may be generated by numerous research procedures. Disposal of this waste at commercial facilities is costly and each dollar spent in disposal costs is money not spent on research. An alternative to commercial disposal is release of radioactive aqueous waste into the sanitary sewer as provided by the Code of Federal Regulations (10CFR20.2003). This method of disposal must meet certain criteria regarding the amount and concentration of radioactivity released to the sewer. In addition, the material must be "readily soluble (or readily dispersible biological material) in water." This paper describes the process used at our R&D facility to determine the solubility of aqueous waste. PMID- 11480860 TI - Photon shielding for a positron emission tomography suite. AB - This paper provides information on the effects of distance and attenuation in lead sheet and gypsum board of the 0.511 MeV photon produced by positron annihilation. Exposure rates are projected external to an adult injected with 185 MBq (5 mCi) of 18F in a fluorodeoxyglucose solution and for the same activity in a small unshielded container. These data have been applied to estimate the shielding requirements for the Positron Emission Tomography (PET) suite operated by the Nuclear Medicine Department of Our Lady of the Lake Regional Medical Center. To assure that exposures are as low as reasonably achievable, lead was added to the walls of the room where the 18F is stored, handled, and injected into the patients. The PET scanner is installed in a room that formerly contained a Computerized Axial Tomography scanner; the existing 1.6 mm of lead sheet was left in place even though it is not required for personnel protection. During the initial phase of operation, a shield test program was conducted to estimate annual exposures to personnel inside and outside the suite. Projection of measured rates over a year of operation demonstrate that whole body doses are well below regulatory limits. PMID- 11480861 TI - A defense in depth approach to radiation protection for 125I production activities. AB - Not all operational radiation protection situations lend themselves to simple solutions. Often a Radiation Protection Program must be developed and implemented for difficult situations. A defense in depth approach to radiation protection was developed for 125I production activities. Defense in depth relies on key radiation protection elements that tend to be mutually supportive and in combination provide reasonable assurance that the overall desired level of protection has been provided. For difficult situations, defense in depth can provide both a reasonable and appropriate approach to radiation protection. PMID- 11480862 TI - Radioactive patient waste at landfills--regulatory impasse. PMID- 11480863 TI - Estimation of radionuclide content in contaminated laundry. AB - Radioactively contaminated laundry is normally sent off site for processing. Laundry is defined as radiologically contaminated anti-cs and respirators. This laundry is shipped as "limited quantity," in accordance with 49CFR173.421. This requires that 95% of the radionuclides shipped are characterized and quantified. In addition, the total quantity must be 10(-3) below the A2 limits specified in 49CFR173. In any facility evaluated, the most conservative (highest activity) waste stream was used as the source term. If a new waste stream is established for a facility, its normalized activity should be compared to the evaluated waste stream to ensure the limits are not exceeded. This article documents a method used for estimating the radionuclide content in contaminated laundry. The maximum values were compared to 49CFR173. Itwas determined that if the contaminated laundry/respirators are shipped in an Interstate Nuclear Services (INS), L-59, limited quantity shipping container and the highest contact radiation level on any side, as measured with an ion chamber, does not exceed 0.5 mR h(-1), the container complies with the requirements of 49CFR173 and could be shipped "limited quantity" from any of the facilities evaluated. PMID- 11480864 TI - Conducting subsurface soil and groundwater radiological investigations: a case study from the University of Rochester. AB - The University of Rochester performed subsurface soil and groundwater radiological assessments in the vicinity of a research building to investigate for contamination from possible past releases of licensed radioactive materials. During the course of this investigation, our contractors developed candidate well drilling and sampling plans. The University of Rochester selected one of these plans, which was subsequently modified due to unexpected costs associated with the local geology. This paper describes the factors that were considered during the development and implementation of the drilling and sampling plan, as well as other considerations and concepts that are inherent in many environmental investigations. PMID- 11480865 TI - Ask the expert. X-ray baggage scanning machines. Neutron well logging. 90Sr into the environment. PMID- 11480866 TI - What have you learned as a radiation safety officer? Interview with Kenneth L. Mossman by R Michel and KL Miller. PMID- 11480867 TI - Exclusion, exemption and clearance...do we need them all? PMID- 11480868 TI - Optically stimulated luminescence dosimeters. PMID- 11480869 TI - Uncertainties in retrospective radon exposure of glass: possible effects of hydration and of leaching. AB - Embedded 210Pb in glass results from exposure to 222Rn and its prompt decay products and is used to estimate integrated, retrospective radon exposures for times up to the 32-y mean life of 210Pb. Certain established uncontrollable factors lead to rather large scatter in the results. We review briefly the known sources that contribute to this scatter and then point out that effects of water and, in particular, hydration layers on glass are probably further sources of error, possibly of major importance, but subject to straightforward checking to reject glass compositions that are unreliable. PMID- 11480870 TI - Analysis of aerosol distribution inside the object "shelter" at the Chernobyl nuclear reactor site. AB - Results of the aerosol sampling inside the Object Shelter at the Chornobyl nuclear reactor site are considered in the work. The results show a consistent and significant contribution of submicron fraction to the total aerosol activity. Different methods of interpretation of impactor data series are compared in order to indicate and detect in the most effective way the submicron fraction as a separate component of the distribution. The methods used are as follows-method of probabilistic-logarithmic scale, method of likelihood function, non-linear approximation by sum of several log-normal distributions, and restoration of the initial distribution using passage functions of impactor cascades. Practically all the collected data on aerosol distribution inside the Shelter show a distinct submicron component. The contribution of the submicron component into the total aerosol activity is about 35%. Influence of the actual aerosol distribution on the dose coefficients is analyzed. PMID- 11480871 TI - EPR tooth enamel dosimetry: optimization of the automated spectra deconvolution routine. AB - A computer routine was developed for automatic deconvolution of electron paramagnetic resonance spectra of tooth enamel samples for individual radiation dose reconstruction in the low dose region. The deconvolution routine uses the non-linear least square fit of a model simulating a tooth enamel spectrum by superposition of derivative Gaussian functions to obtain the amplitude of the dosimetric radiation induced signal. The parameters of the model and of the routine were optimized on a dose response level using a criterion of the least standard deviation of the derived radiation induced signal amplitude from the regression line vs. the nominal doses for the series of spectra of samples irradiated in known doses in the range 0-500 mGy. It was found that for the series of spectra of the heterogeneous samples (every sample is prepared from different teeth of different persons), it is essential to vary in the least square fit the parameters describing the shape of the native background signal in order to obtain the best accuracy. In the case of the series of spectra of the homogeneous (pooled) samples, almost the same accuracy of the results was obtained using the procedures with varied and fixed background signal parameters. The standard error of the dose reconstruction by the optimized deconvolution procedure was estimated as about 30 mGy for heterogeneous samples and 20 mGy for homogeneous samples. PMID- 11480872 TI - A theoretical evaluation of the assessment of effective dose using multiple personnel dosimeters. AB - The ability of a dose calculation algorithm, using the readings of multiple dosimeters, to accurately assess the effective dose under different photon irradiation conditions was assessed using computer simulation. The algorithm was that described in American National Standards Institute publication N13.41. Monte Carlo calculations with an anthropomorphic humanoid phantom were used to calculate the effective doses and also the expected readings of the multiple dosimeters. The irradiation geometries considered included a point source placed at several locations at a distance of 100 cm in front of the phantom, as well as an anterior-posterior plane parallel beam with a lead shield interposed between the phantom and the source. The point source energies considered were 0.05, 0.6, and 2 MeV, and the beam energy was varied between 0.03 and 10 MeV. Also considered were the estimates of effective dose based on the highest reading of the multiple dosimeters, a practice that is currently used in many work places. The results showed that use of the algorithm resulted in substantial improvements in the ability to accurately estimate effective dose. However, the results also showed that the improvements in accuracy were achievable only by using a calibration factor for the dosimetry that is different from the one obtained in current dosimetry calibration practices, and that without the use of this factor, the algorithm tended to underestimate the effective dose for nearly all the irradiation geometries considered. In addition, it appeared that this calibration factor is not constant but varies with irradiation conditions. There thus appears to be a problem of proper dosimetry calibration for use with the algorithm. This work considered only anterior posterior irradiations, and additional work is needed to assess the performance of the algorithm in other non-uniform irradiation geometries. PMID- 11480873 TI - 137Cs concentration in the environment of Kaiga of south west coast of India. AB - This paper presents the results of 137Cs activity in soil and vegetation obtained through a carefully planned pre-operational survey for the establishment of baseline data on background radiation level and distribution of radionuclides in the environment of Kaiga, in the south west coast of India, where two nuclear power reactors of 235 MWe each are under construction. The vertical profile in soil and the seasonal variation in the activity have been studied. Nine commonly available vegetation species of Kaiga region were analyzed for their 137Cs activity, and the soil-to-plant transfer coefficients were evaluated. Measurements were made by gamma spectrometry method employing a 90 cc HpGe detector and a 3 x 3-inch well type NaI(Tl) detector. Results show significant levels of 137Cs activity in soil samples--ranging from below detection limit (BDL) to 65.4 Bq kg(-1). The mean value of activity observed in 0-5 cm soil profiles of Kaiga environs is significantly higher when compared to the reported values for other environs of India. Depth profile studies show that the 137Cs activity decreases with the increasing soil depth, and it is below detection limit for depths more than 25 cm. The mean value of effective dose commitment due to the presence of 137Cs in soil was 167.2 microSv. Vegetation samples of the region show elevated levels of 137Cs concentration. Epiphytic plants Cymbidium aloifolium (Lo.) Swartz. and Petrobryopsis tumida (Hook.) Dix. have been identified as bio-indicators to monitor the fallout of this radionuclide in the environment of Kaiga. Seasonal variation studies indicate higher fallout of this radionuclide along with rain. PMID- 11480874 TI - Quality control of mitigation methods for unusually high indoor radon concentrations. AB - The present study's objective was to control the quality of different mitigation methods for unusually high indoor radon (222Rn) concentrations of up to 274,000 Bq m(-3) in a village (Umhausen, 2,600 inhabitants) in western Tyrol, Austria. Five years after mitigation, five different remedial actions were examined on their quality by means of measuring indoor radon concentrations with charcoal liquid scintillation radon detectors and with a continuously recording AlphaGuard detector. Mitigation method in house 1--a mechanical intake and outlet ventilation system with heat exchanger in the basement, combined with a soil depressurization system--was characterized by long-term stability. With most favorable air pressure (+100 Pa) in the basement, mean basement radon concentrations in the winter were reduced from 200,000 Bq m(-3) to 3,000 Bq m(-3) by this method 5 y after mitigation. Acting against experts' instructions, the inhabitants had switched off the ventilation system most of the time to minimize power consumption although it had been proven that ventilation reduced mean basement radon concentration by a factor of about 3 in the winter and about 15 in the summer. Mitigation method in house 2-soil depressurization with two fans and loops of drainage tubes to withdraw radon from the region below the floor and outside the basement walls, and from soil below that part of the house with no basement-had been the most successful remedial measure until the winter of 1999 (i.e., 6 y after mitigation), when micro-cracks opened and consequently mean basement radon concentration increased from 250 Bq m(-3) to 1,500 Bq m(-3). Measures to block these microcracks and to minimize soil drying are being developed. Five years after mitigation, the remedial method used in house 3--a multilayer floor construction, where a fan was used to suck radon from a layer between bottom slab and floor-reduced winter mean radon concentration from 25,000 Bq m(-3) to 1,200 Bq m(-3), with the ventilation on and the basement door open. Mitigation method in house 4--a basement sealing technique--was unsuccessful with almost identical radon concentrations during all the five years since mitigation had started. Mitigation method in house 5--a waterproof basement technique especially for future homes--reduced mean basement radon concentration below 300 Bq m(-3) and mean ground floor radon concentration below 200 Bq m(-3), which is the Austrian action level for newly constructed buildings. These findings indicate that even in areas with extremely high radon concentrations, effective mitigation of indoor radon can be achieved provided that house-specific long term, stable mitigation techniques are applied. PMID- 11480875 TI - Conversion coefficients based on the VIP-Man anatomical model and EGS4. AB - A new set of conversion coefficients from kerma free-in-air to absorbed dose and kerma free-in-air to "effective VIP-Man dose" has been calculated for external monoenergetic photon beams from 10 keV to 10 MeV using an image-based whole-body anatomical model. This model, called VIP-Man, was recently developed at Rensselaer from the high-resolution color images of the National Library of Medicine's Visible Human Project. An EGS4-based Monte Carlo user code, named EGS4 VLSI, was developed to efficiently process the extremely large image data in VIP Man. Irradiation conditions include anterior-posterior, posterior-anterior, right lateral, left lateral, rotational, and isotropic geometries. Conversion coefficients from this study are compared with those obtained from two mathematical models, ADAM and EVA. "Effective VIP-Man doses" differ from the previously reported effective dose results by 10%-50% for photons between 100 keV and 10 MeV. Discrepancies are more significant at lower energies and for individual organ doses. Since VIP-Man is a realistic model that contains several tissues that were not previously defined well (or not available) in other models, the reported results offer an opportunity to improve the existing dosimetric data and the mathematical models. PMID- 11480876 TI - Fluence-to-dose conversion coefficients based on the VIP-Man anatomical model and MCNPX code for monoenergetic neutrons above 20 MeV. AB - A new set of fluence-to-absorbed dose and fluence-to-effective dose conversion coefficients has been calculated for high-energy neutrons using a whole-body anatomical model, VIP-Man, developed from the high-resolution transversal color photographic images of the National Library of Medicine's Visible Human Project. Organ dose calculations were performed using the Monte Carlo code MCNPX for 20 monoenergetic neutron beams between 20 MeV and 10,000 MeV under 6 different irradiation geometries: anterior-posterior, posterior-anterior, left lateral, right lateral, isotropic, and rotational. For neutron Monte Carlo calculations, results based on an image-based whole-body model were not available in the literature. The absorbed dose results for 24 major organs of VIP-Man are presented in the form of tables and selected figures that compare with those based on simplified mathematical phantoms reported in the literature. VIP-Man yields up to 40% larger values of effective dose and many organ doses, thus suggesting that the results reported in the past may not be conservative. PMID- 11480877 TI - Organ dose conversion coefficients for 0.1-10 MeV electrons calculated for the VIP-Man tomographic model. AB - A whole-body tomographic model, called VIP-Man, was recently developed at Rensselaer Polytechnic Institute from the high-resolution color photographic images of the National Library of Medicine's Visible Human Project. An EGS4-based Monte Carlo user code, named EGS4-VLSI, was developed to efficiently transport electrons using the large image data set for VIP-Man. VIP-Man has been used to calculate doses for neutrons and photons. This paper presents a new set of fluence-to-absorbed-dose conversion coefficients for monoenergetic electron beams between 100 keV and 10 MeV for VIP-Man. Irradiation conditions include anterior posterior, posterior-anterior, right lateral, left lateral, rotational, and isotropic source geometries. Comparisons between organ doses from VIP-Man, which is taller and heavier than the Reference Man, and existing data from mathematical models show significant discrepancies. It appears that even slight differences between body models can cause dramatic dosimetric deviations for low penetrating electron irradiation. This suggests that a single standard body model may poorly represent a large population and may not be acceptable for electron dosimetry. PMID- 11480878 TI - External air kerma rate measurements--comment on paper by Narayana et al. PMID- 11480879 TI - The definition and meaning of treatment-resistant depression. AB - Most patients treated for an episode of unipolar or bipolar major depression are treatment resistant in the sense that the majority do not achieve full remission with the first somatic or psychosocial treatment they receive. Little attention has been given to formalizing criteria for evaluating the nature and extent of treatment resistance, even though determining the adequacy and outcome of prior treatment trials is key in clinical decision making about subsequent treatment. Furthermore, determining the adequacy of prior treatment is essential since substantial evidence indicates that large numbers of depressed patients are undertreated, resulting in prolonged episodes and the appearance of "pseudoresistance." Adequacy of antidepressant treatment trials should be defined in terms of thresholds for the dosage and duration of medication, adherence, and clinical outcome. The Antidepressant Treatment History Form is presented as one method to formalize the evaluation of treatment adequacy and treatment resistance. PMID- 11480880 TI - Clinical features of treatment-resistant depression. AB - As many as 30% to 40% of patients with major depressive disorder are unresponsive to a trial of antidepressant medication. Many patients labeled with treatment resistant depression actually have pseudoresistance, in that they have been inadequately treated or are misdiagnosed. Others may have unrecognized comorbid psychiatric or general medical conditions that contribute to treatment resistance. Variables such as gender, family history, age at onset, severity, and chronicity have also been evaluated as possible risk factors for treatment resistant depression. This article reviews the current literature regarding the clinical characteristics of treatment-resistant depression, with particular attention to the relevance of these factors for clinical decision making. PMID- 11480881 TI - The burden of disease for treatment-resistant depression. AB - Assessing the consequences of specific diseases on global, national, and individual levels is complex. The Global Burden of Disease Study was launched in 1992 to develop objective measures of the burden of disease. Two measures have become widely accepted: disability-adjusted life-years (DALYs) assesses years of life lost due to a disease plus years lived with the disability due to that disease, and years lived with disability (YLDs) is a related measure with greater relevance for diseases that do not routinely produce earlier mortality. When DALYs and YLDs were compared world-wide for 100 disorders, they revealed a huge burden of disease for depression. Indeed, the findings were startling. Neuropsychiatric conditions are by far the world's leader in YLDs, accounting for almost 30%. Unipolar major depressive disorder alone accounted for 11% of global YLDs. The disability of major depressive disorder produces its greatest burden upon women and starts early in life. No separate disability assessments have been compiled for treatment-resistant depression, but of individuals with major depressive disorder, the most severely disabled are those with treatment resistant depression. The contributions to the morbidity associated with major depressive disorder and treatment-resistant depression include widespread prevalence; relatively early symptom onset; severe underdiagnosis and undertreatment; genetic vulnerabilities and precipitation or accentuation by relatively unavoidable stressors; a longitudinal pattern of frequent recurrences with increasing frequency, severity, and consequences unless treated with maintenance strategies; inadequate prioritization of recurrence prevention among clinicians; and possible suppression of brain neurogenesis, neuronal atrophy, cell death, hippocampal dysfunction, and magnetic resonance imaging changes for those with chronic treatment-resistant depression. Since the patterns of recurrences, cycle acceleration, and increasing severity of treatment-resistant depression are key reasons for its high burden, reducing the burden requires an entire paradigm shift, including emphasis on the prevention of recurrences. Only then will this prevalent, disabling yet treatable disorder lose its ignominious status as a world leader in disease burden. PMID- 11480882 TI - Definitions of antidepressant treatment response, remission, nonresponse, partial response, and other relevant outcomes: a focus on treatment-resistant depression. AB - Multiple definitions have been used to characterize the outcome of treatments for depression. Beyond the simple criterion of a statistically significant improvement in depression rating scale scores, researchers have had to use more clinically relevant categorical outcomes: response (without remission), remission, nonresponse, partial response, relapse, recurrence, recovery, and, more recently, depressive breakthrough. This article reviews the definitions of these terms and their relevance for the study of treatment-resistant depression. PMID- 11480883 TI - RP-1776, a novel cyclic peptide produced by Streptomyces sp., inhibits the binding of PDGF to the extracellular domain of its receptor. AB - RP-1776, a novel cyclic peptide, was isolated from the culture broth of Streptomyces sp. KY11784. RP-1776 selectively inhibited the binding of PDGF BB to the extracellular domain of the PDGF beta-receptor with an IC50 value of 11 +/- 6 microM. Detailed binding experiments suggested that RP-1776 directly interacts with PDGF BB. RP-1776 inhibited the phosphorylation of the PDGF beta-receptor induced by PDGF BB. These results suggested that RP-1776 antagonizes the signaling of PDGF BB probably through the inhibition of PDGF BB binding to the PDGF beta-receptor. PMID- 11480884 TI - S-15183a and b, new sphingosine kinase inhibitors, produced by a fungus. AB - In the course of our screening for inhibitors of sphingosine kinase, we found two active compounds in a culture broth of a fungus, Zopfiella inermis SANK 15183. The structures of the compounds, named S-15183a and b, were elucidated by a combination of spectroscopic analyses to be new azaphilone-type metabolites. S 15183a and b inhibited sphingosine kinase from rat liver with IC50 values of 2.5 and 1.6 microM, respectively. S-15183a also inhibited endogenous SPH kinase activity in intact platelets. PMID- 11480885 TI - TMC-69, a new antitumor antibiotic with Cdc25A inhibitory activity, produced by Chrysosporium sp. TC1068. Taxonomy, fermentation and biological activities. AB - A new antibiotic designated TMC-69 has been isolated from the fermentation broth of a fungal strain Chrysosporium sp. TC 1068. TMC-69 exhibited moderate in vitro cytotoxic activity. TMC-69-6H, a derivative of TMC-69 prepared by hydrogenation, possessed more potent in vitro cytotoxicity than TMC-69, and exhibited in vivo antitumor activity against murine P388 leukemia and B16 melanoma. TMC-69-6H was found to specifically inhibit Cdc25A and B phosphatases. PMID- 11480886 TI - Streptocidins A-D, novel cyclic decapeptide antibiotics produced by Streptomyces sp. Tu 6071. I. Taxonomy, fermentation, isolation and biological activities. AB - Four novel cyclic homodecapetide antibiotics, streptocidins A-D were detected in the mycelium extract of Streptomyces sp. Tu 6071 by HPLC-diode-array and HPLC electrospray-mass-spectrometry screening. The peptides which were closely related in structure to the tyrocidines and gramicidins of Bacillus brevis show antibiotic activities against Gram-positive bacteria. PMID- 11480887 TI - Streptocidins A-D, novel cyclic decapeptide antibiotics produced by Streptomyces sp. Tu 6071. II. Structure elucidation. AB - The structures of the new antibiotics streptocidins A approximately D were elucidated as cyclic decapeptides cyclo[L-Val1-L-Orn2-L-Leu3-D-Phe4-L-Pro5-L-Leu6 X7-L-Asn8-L-Gln9-X10] with X7=D-Trp (A, B, C) or D-Phe (D) and X10=L-Tyr (A), L Trp (B, D), or D-Trp (C). The amino acid composition (including the configuration) of the substances was determined by chiral-phase GC-MS of the hydrolysates. The sequences were established by EDMAN degradation following linearisation of the cyclic peptides upon treatment with LiAlH4. NMR spectroscopic studies of streptocidins C and D confirmed the proposed sequences and provided conformational data which indicate a molecular topology of streptocidins C and D similar to those of tyrocidine A and gramicidin S. PMID- 11480888 TI - TPU-0031-A and B, new antibiotics of the novobiocin group produced by Streptomyces sp. TP-A0556. AB - Two novel antibiotics, TPU-0031-A and B, were isolated from the culture broth of an actinomycete strain. The producing strain, TP-A0556, was identified as Streptomyces sp. based on the taxonomic study. The new antibiotics were obtained by solvent extraction and chromatographic purification. Spectroscopic analyses showed that TPU-0031-A and B were 7'-demethylnovobiocin and 5" demethylnovobiocin, respectively. These compounds showed antibiotic activity against Gram-positive and -negative bacteria. PMID- 11480889 TI - Biotransformation of selamectin with Streptomyces lydicus SX-1298 using a novel static agar fermentation system with Reemay mesh. AB - A novel approach to biotransformation is described using a solid medium matrix and Reemay mesh that gives efficient biotransformation of compounds with minimal matrices in the ensuing gum solids. Using this approach with a newly isolated biotransforming organism, Streptomyces lydicus SX1298, a series of hydroxylations and an O-demethylation is described for selamectin the first endectocide for cats and dogs. PMID- 11480890 TI - Synthesis of hydrophobic N'-mono and N',N"-double alkylated eremomycins inhibiting the transglycosylation stage of bacterial cell wall biosynthesis. AB - A series of hydrophobic N'-mono and N',N"-double alkylated derivatives of the glycopeptide antibiotic eremomycin were synthesized by reductive alkylation after preliminary protection of the N-terminal amino group of the peptide backbone. The investigation of the antibacterial activity in vitro showed that N'-C10H21- and N'-p-(p-chlorophenyl)benzyl derivatives of eremomycin are the most active against vancomycin-resistant enterococci among the compounds obtained though they are less effective than the corresponding lipophilic derivatives of vancomycin. The introduction of two hydrophobic substituents led to a decrease in activity against both susceptible and resistant bacteria. The biochemical evaluation of the mode of action revealed that in addition to binding to D-Ala-D-Ala these compounds also have an alternative mechanism of action that does not require substrate binding. PMID- 11480891 TI - New cephalosporin antibiotics with 3-triazolylpyridiniummethyl substituents. PMID- 11480892 TI - Anti-angiogenic activities of novel isocoumarins, AGI-7 and sescandelin. PMID- 11480893 TI - Effects of combining public assistance and employment on mothers and their young children. AB - One goal of recent welfare reform legislation is to move welfare-dependent mothers with young children into the paid labor force. However, prior to the new legislation, many welfare-dependent women were already engaged in employment activities. In this paper we examine whether child or maternal well-being is influenced by a mother's strategy of combining work and public assistance receipt in the late 1980s. Measures of well-being include children's cognitive test scores and behavior problems, parenting behavior, and maternal mental health, social support, and coping strategies collected when children were 2 1/2 to 3 years of age. Data from the Infant Health and Development Program (a sample of low birthweight, premature infants born in 8 sites in 1985) were used to identify low-income families (incomes under 200% of the poverty threshold; N = 525). Comparisons were made among mothers in the following groups: (a) Work Only, (b) Some Work-Some Welfare, (c) Some Work-No Welfare, (d) No Work-No Welfare, and (e) Welfare Only. Mothers in the Some Work-Some Welfare group had children with cognitive and behavioral scores similar to children whose mothers were in the Work Only group; these two groups also had similar mental health, social support, and coping scores. However, not working and receiving welfare (Welfare Only) was associated with negative cognitive and behavioral outcomes for children, with less stimulating home learning environments, lower maternal mental health, less social support, and more avoidant coping strategies. We discuss the proposition that welfare and work may be complementary rather than opposing strategies, in terms of putting together a family income package. PMID- 11480894 TI - Maternal depressive symptoms and low literacy as potential barriers to employment in a sample of families receiving welfare: are there two-generational implications? AB - This study examines the role of maternal depressive symptoms and low maternal literacy in predicting outcomes in two generations in families receiving welfare: mothers' employment and children's development. The sample consists of 351 African-American families, each with a preschool-age child, in which the mother had applied for or was receiving welfare. Close to the start of the study, 52.6 percent of the mothers in the sample had scores indicating lower literacy, 39.5 percent reported moderate to high levels of depressive symptoms, and 24.6 percent had a co-occurrence of these. Using continuous scores, in multivariate analyses of variance, neither level of literacy, extent of depressive symptoms, nor the interaction of these, were found to predict two measures of subsequent employment (any employment across the two year follow-up period, and current employment at the time of the follow-up). However, when cut points were used (low literacy; moderate to high depressive symptoms), mothers with low literacy were found less often to be employed approximately two years later. Multivariate analyses of variance examining the set of child outcomes (cognitive school readiness and behavior problems) in light of mothers' depressive symptoms and literacy level found a statistically significant interaction of literacy level and extent of depressive symptoms: children of mothers with more depressive symptoms had less favorable developmental outcomes only in the presence of low maternal literacy. Structural equation models provide evidence that parenting behavior mediates the relationship between the predictor variables and child outcomes, and that the pathways from depressive symptoms through parenting to child outcomes are stronger when maternal depressive symptoms co-occur with low maternal literacy. PMID- 11480895 TI - Effects of welfare reform on children of adolescent mothers: moderation by maternal depression, father involvement, and grandmother involvement. AB - There are few data available on how effects of welfare reform on children may differ for different kinds of families. This study examined the constructs of maternal depression, father involvement, and grandmother involvement as moderators of the effects of a welfare reform program on children. The sample included 1,602 children between the ages of 3 and 6 1/2 whose mothers had participated in New Chance, a multi-site welfare-to-work program with an emphasis on human capital development (i.e., education, job training). Father and grandmother co-residence were investigated, in addition to perceived support from these family members, in predicting cognitive and behavioral child outcomes. The mediating roles of parenting stress and emotional support in parenting were also explored. Although main effects were found for the intervention and for maternal depression, no main effects were found for the family ecology variables. However, interactions suggested complex relationships among New Chance and the family characteristics for both the full sample and for Latina families. Results are discussed in terms of policy implications and possible future research. PMID- 11480896 TI - What is your diagnosis? Retained surgical sponge. PMID- 11480897 TI - Evaluation of methods of platelet counting in the cat. AB - Platelet counts were performed in 50 cats presented for diagnostic investigation. For each cat, counts were obtained using a manual haemocytometer method and compared with counts obtained by estimation from a stained blood smear, a QBC VetAutoread analyser, a Zynocyte VS/2000 analyser, impedance automated counts on a Baker System using both EDTA and citrated anticoagulated blood, and use of a Zynostain modified counting chamber kit. None of the methods gave high correlation with the haemocytometer counts. The blood smear estimation of platelet counts had the highest correlation (r = 0.776) and was the only method to have reasonable values for both sensitivity and specificity. With the impedance automated counts, citrated anticoagulated blood had marginally higher correlation than EDTA anticoagulated blood, and the time between blood sampling and platelet count determination had no effect on the count obtained. When in house analyser or impedance automated platelet counts are abnormal or not consistent with clinical findings, the authors recommend that a manual platelet count using either haemocytometry or examination of a blood smear is performed. PMID- 11480898 TI - 'Battered pets': sexual abuse. AB - A study of non-accidental injury in small animals in the UK, based on responses from a random sample of small animal practitioners, identified 6 per cent of the 448 reported cases as being sexual in nature. Twenty-one cases occurred in dogs, five in cats and two in unspecified species. Reasons for suspecting sexual abuse were: the type of injury; behaviour of the owner; statements from witnesses; and admission by the perpetrator. Types of injury included vaginal and anorectal penetrative (penile and non-penile) injury, perianal damage, and trauma to the genitals. Some injuries (such as castration) were extreme, and some were fatal. In contrast, other cases revealed no obvious damage. The type and severity of injuries were similar to those described in texts on child abuse and human forensic pathology. PMID- 11480899 TI - Congenital unilateral cleft of the soft palate in six dogs. AB - Unilateral cleft of the soft palate occurs sporadically in dogs, and there are few reports in the veterinary literature regarding its aetiology and management. This report describes six dogs with the condition. The dogs' primary presenting problem was rhinitis. The defects were successfully repaired using a two-layer closure technique with absorbable suture in five of the cases. Surgical correction was not performed in the remaining case. PMID- 11480900 TI - Urethral entrapment following pelvic fracture fixation in a dog. AB - An 18-month-old female crossbred dog was presented with a unilateral sacroiliac luxation and separation of the pelvic symphysis. Surgical correction of the luxation with screw fixation led to entrapment of the urethra between the symphyseal parts of the two hemipelves. PMID- 11480901 TI - Primary ciliary dyskinesia with situs inversus totalis, hydrocephalus internus and cardiac malformations in a dog. AB - A nine-month-old golden retriever bitch was presented with exercise intolerance and recurrent nasal discharge. Based on clinical, radiographic and ultrasonographic examination, a diagnosis of rhinitis, situs inversus totalis and tricuspid valve insufficiency was established. The results of video- and electron microscopy studies of the respiratory epithelium were compatible with primary ciliary dyskinesia (PCD). However, no evidence of a primary ultrastructural defect of the cilia was found. The dog was euthanased because of the poor prognosis. At necropsy, a hydrocephalus internus and a subaortic stenosis were additionally diagnosed. PCD, in combination with situs inversus, has been previously reported in golden retrievers, but without a concomitant hydrocephalus internus. Furthermore, concomitant occurrence of internal cardiac malformation and PCD has not previously been reported in the dog. PMID- 11480902 TI - Acute respiratory failure caused by leukaemic infiltration of the lung of a dog. AB - A seven-year-old crossbred male dog with a suspected leukaemic condition was referred for investigation and treatment. A bone marrow aspirate revealed an acute myeloid leukaemia. Combination chemotherapy was administered and the dog initially improved, but 18 days after the initiation of therapy its body condition deteriorated and the animal developed acute respiratory distress. On postmortem examination, extensive leukaemic pulmonary infiltrates were evident. PMID- 11480903 TI - Discospondylitis associated with three unreported bacteria in the dog. AB - Three cases of discospondylitis in the dog, caused respectively by Pseudomonas aeruginosa, Enterococcus faecalis and Staphylococcus epidermidis, are reported. These bacteria have not previously been documented as being the cause of the condition in this species. The neurological involvement was severe in all three cases. Two dogs were paraplegic and one was tetraplegic, and one of the paraplegic dogs had no deep pain on the extremities of both pelvic limbs. All dogs were treated surgically with a satisfactory neurological recovery. PMID- 11480904 TI - Subluxation/luxation of the second carpal bone in two racing greyhounds and a Staffordshire bull terrier. AB - Subluxation of the second carpal bone (C2) in two racing greyhounds, and luxation associated with other carpal injuries in a Staffordshire bull terrier, were diagnosed as causes of thoracic limb lameness. The clinical signs of subluxation were subtle, with local soft tissue swelling and a pain reaction on digital pressure over the dorsal aspect of C2. In contrast, the dog with a complete luxation was very lame, having marked soft tissue swelling with carpal hyperextension and valgus. Diagnosis was made by radiography. Subluxation of C2 was seen as a small, dorsally displaced opacity between the proximal and distal rows of carpal bones on the mediolateral view and, on the oblique view taken in only one case, as a marked dorsal displacement of one-third of the proximal joint surface of C2. In the case of luxation, C2 appeared on the mediolateral view lying dorsal to the radial carpal bone. There was also fracture of C4. Both greyhounds were treated by internal fixation and returned to racing. The Staffordshire bull terrier was treated by pancarpal arthrodesis with a successful outcome. PMID- 11480905 TI - Malignant seminoma with systemic metastases in a dog. AB - A three-year-old male crossbred dog was presented with multiple cutaneous nodules on the dorsum. Unilateral testicular enlargement ensued. Histopathological examination of testicular tissue and the skin lesions revealed a malignant seminoma. Despite the initiation of adjuvant chemotherapy, the dog died three months later due to systemic metastases of the seminoma to the skin, scrotum, eyes, liver, kidney and peritoneum. PMID- 11480906 TI - Effect of long-term intermittent periodontal care. PMID- 11480907 TI - BSAVA's response to the IRG's report on dispensing. PMID- 11480908 TI - Identifying the genes responsible for inherited diseases in dogs. PMID- 11480909 TI - Equality of the sexes: mammalian dosage compensation. AB - X chromosome inactivation achieves dosage equivalence for most X-linked genes between the two X chromosomes in females and the single X chromosome in males. In this article the evidence for random inactivation of an X chromosome is reviewed, along with the exceptions that result in nonrandom inactivation. Another exception to X chromosome inactivation is the presence of genes that escape inactivation and are expressed from both the active and inactive X chromosomes. The phenotypic consequences of such expression from the inactive X chromosome are discussed. The major players in the process of inactivation are presented. Initiation of inactivation requires the functional RNA, XIST, and the subsequent stable inactivation of the X chromosome relies upon the recruitment of many other factors, the majority of which are generally associated with heterochromatin. PMID- 11480910 TI - X-Y translocations and sex differentiation. AB - Translocations involving the X and Y chromosomes are often associated with anomalies of gonadal development. Transfer of Yp sequences, including the testis determining SRY gene, to the terminal portion of the short arm of the X chromosome is associated with 46,XX maleness and in rare cases 46,XX true hermaphroditism. Three classes of XX males have been defined on the basis of the extent of Y material transferred to the X chromosome. In one class, the transfer of material involves aberrant recombination between two highly homologous genes, PKRX and PKRY, and there is evidence to suggest that this interchange is influenced by the Y chromosome background. Other types of X-Y translocations associated with anomalies of sex differentiation include Xp-Yq translocations, which result in a functional disomy of Xp sequences including the DSS locus and are associated with 46,XY complete or partial gonadal dysgenesis. In rare cases Yp-Xq translocations have been described in association with 46,XX maleness. PMID- 11480911 TI - Molecular analysis of genes on Xp controlling Turner syndrome and premature ovarian failure (POF). AB - Monosomy X has been known to be the chromosomal basis of Turner syndrome (TS) for more than four decades. A large body of cytogenetic data indicates that most TS features are due to reduced dosage of genes on the short arm of the X chromosome (Xp). Phenotype mapping studies using molecular cytogenetic and genetic techniques are beginning to localize the Xp genes that are important for various TS features, and a comprehensive catalog of candidate genes is becoming available through the Human Genome Project and related research. It is now possible to assess the contributions of individual genes to the TS phenotype by mutational analysis of karyotypically normal persons with specific TS features. This strategy has succeeded in identifying a gene involved in short stature and is being applied to premature ovarian failure and other TS phenotypes. PMID- 11480912 TI - Ring X and other structural X chromosome abnormalities: X inactivation and phenotype. AB - Patients who carry a structural abnormality of the X chromosome are a fascinating group who have provided opportunities to evaluate genotype/phenotype correlation in relation to X chromosome content and inactivation. Turner syndrome (TS) is most commonly associated with a 45,X karyotype and presents with an array of phenotypes, the main ones being poor viability in utero, ovarian failure and infertility, short stature, lymphedema, and other congenital malformations but usually not mental retardation. In some TS patients the karyotype shows both a normal X and a structurally rearranged X chromosome. These structural abnormalities, which include deletions, duplications, inversions, translocations, and rings, are associated with chromosome breaks and significant imbalance of gene content of the X chromosome. However, such abnormalities are generally well tolerated because of the preferential inactivation of the abnormal X, which can restore, at least in part, a balanced genetic makeup. This beneficial effect of X inactivation results in a mild phenotype in most patients with structural abnormalities of the X, similar to that found in TS patients with a 45,X karyotype. However, in cases of ring X chromosomes and of X/autosome translocations the incidence of mental retardation and other congenital abnormalities can be significantly higher than in TS. These abnormal phenotypes can be ascribed to failed or partial X inactivation and/or incomplete selection in favor of cells with normal balance of gene expression. In this article, we present phenotype/genotype correlation in female patients with structural abnormalities of the X and address the role of X inactivation and cell selection in the phenotypic findings. Our review emphasizes a subset of rare patients with ring X chromosomes who have provided evidence of a direct role for X inactivation in determining phenotypes. PMID- 11480913 TI - The female and the fragile X reviewed. AB - Nearly 15 years ago, female carriers of the fragile X mental retardation syndrome were noted to have an increased incidence of twin pregnancies. Since then, much evidence has accumulated supporting the notion of ovarian dysfunction in fragile X carriers, in the forms of increased dizygotic twinning and premature ovarian failure. However, despite a decade and a half of research regarding this association, the underlying mechanism remains a mystery. This article reviews the population-based studies that have examined this association and discusses possible reasons for the variations in results. In addition, results from more recent studies on endocrine function in fragile X carriers are discussed. These data, when considered in conjunction with our emerging understanding of the molecular biology of the fragile X gene (FMR1) and its protein product (FMRP), are beginning to elucidate possible mechanisms for the association between fragile X syndrome and ovarian dysfunction. PMID- 11480914 TI - Differential hybridization and other strategies to identify novel ovarian genes. AB - Approaches to define patterns of gene expression have applications in a wide range of biological systems. The completion of the human genome project establishes a unique opportunity to understand the molecular control of different biological events through functional analysis. However, the huge database that currently exists will demand the utilization of high-throughput techniques for the assessment of multiple DNA sequences in a rapid and efficient manner. By determining genes with differential expression, it may be possible to decipher the mechanisms that underlie the control of different physiological pathways, which, in turn, may define future strategies to manage them. Although there exist a vast number of tools to screen genes for differential expression, we briefly examine here only some of those with regard to their advantages and disadvantages. PMID- 11480915 TI - Skewed X inactivation and recurrent spontaneous abortion. AB - Recent studies show that women experiencing recurrent spontaneous abortion exhibit nonrandom X-chromosome inactivation (XCI) more often than in controls. This suggests that genetic factors may be important in explaining the losses in this subset of women. Nonetheless there are a number of possible explanations for this finding and the underlying causes may be heterogeneous. One hypothesis commonly cited is that a mutation on the X chromosome results in both preferential inactivation of the mutated X as well as lethality of male embryos inheriting this mutated X. However, this hypothesis does not explain the increase in chromosome abnormalities observed in the karyotyped losses from women with recurrent pregnancy loss and skewed XCI. This finding leads us to suggest that the mechanism involved may be associated with a reduction in number of ovarian follicles, either due to X mutations affecting oocyte atresia or a restriction in precursor pool size during development. PMID- 11480916 TI - Skewed X inactivation in X-linked disorders. AB - X chromosome inactivation is a process by which the dosage of proteins transcribed from genes on the X chromosome is equalized between males (XY) and females (XX) through the silencing of most genes on one of the two X chromosomes in females. Although the choice of which of the two X's is inactivated is entirely random, not all women have a 50:50 ratio of cells with one or the other X chromosomes active. A number of different mechanisms lead to extremely skewed ratios and this can result in expression of the phenotype of X-linked recessive disorders in females. Nonrandom X inactivation patterns are also associated with selective female survival in male-lethal X-linked dominant disorders or with variable severity of the phenotype in women carrying X-linked dominant mutations. These features are important for the study and gene identification of X-linked disorders and for counseling of affected families. PMID- 11480917 TI - Mental development in polysomy X Klinefelter syndrome (47,XXY; 48,XXXY): effects of incomplete X inactivation. AB - The child with XXY or a variant form is a fertile ground for scientific investigation because of the homogeneity of the disorder and the increased prevalence of learning disorders associated with it. However, the research studies of boys with XXY (Klinefelter syndrome) have been plagued by a variety of factors from small sample size, methodological flaws, and ascertainment bias. In spite of these shortcomings, there remains some consistency to the neurobehavioral profile of this disorder. In general, the prenatal population of boys with XXY is less affected developmentally and is more academically successful than the boys identified through postnatal diagnosis. Boys with XXY often have decreased muscle tonus, delayed speech, and language skills with an increased incidence of reading differences and dyslexia. It does appear that social difficulties may be mediated by language processing problems and temperamental issues. The neurobehavioral and neurocognitive phenotype of boys with XXY places them at risk for school failure and secondary behavioral disturbances. Therefore, early evaluation and intervention is strongly recommended since the prognosis may be improved significantly with appropriate therapeutic intervention. The natural history of this disorder is not well defined from the neurodevelopmental, neuroimaging, and cytogenetic perspective. Further investigation into the effects and the relationships between parental origin and outcome may provide many answers regarding the variability of the disorder. Lastly, some promising theories have been postulated regarding the neurobiological etiology of this disorder, which requires more investigation if we are to understand the pathogenesis of this XXY and its effect on learning. PMID- 11480918 TI - Hydroxymethylation and oxidation of Organosolv lignins and utilization of the products. AB - Organosolv lignins obtained from Eucalyptus grandis, sugarcane bagasse and Picea abies by Acetosolv, Formacell and Organocell processes were characterized, fractionated and converted to hydroxymethylated and oxidized products. The reactivity of lignins with formaldehyde did not improve significantly with the fractionation. Both eucalyptus Acetosolv (EAc) and eucalyptus Formacell (EFo) lignins retained high heterogeneity in relation to the molecular weight distribution but not in relation to structural units. The temperatures of the exothermic peaks and the apparent activation energies for the cross-linking are different for hydroxymethylated lignins and phenol, with similar cure temperatures of the resols. Chemical oxidation using cobalt(II) and manganese(II) salts furnished oxidized lignins with improved chelating properties. These chelating agents can remove up to 14% of Mn present in pulps, decreasing the peroxide consumption in the bleaching process. The products obtained can be also used as oxidized phenols and controlled-release matrices. Oxidation of Acetosolv bagasse lignin with polyphenol oxidase furnishes lignins with chelating capacity 110% higher than that of original lignin. PMID- 11480919 TI - Effect of oxygen delignification on the rate and extent of enzymatic hydrolysis of lignocellulosic material. AB - In this study, we examined the effect of oxygen delignification on the rate and extent of enzymatic hydrolysis (using commercial cellulase and beta-glucosidase) of a number of lignocellulosic substrates, including kraft pulp (model substrate), pulp mill primary clarifier sludge (PCS) and steam-exploded Douglas fir chips. Oxygen delignification removed up to 67% of the lignin from softwood pulp and improved the rate of, and yield from, hydrolysis by up to 111% and 174%, respectively. Glucose yield varied linearly with fractional lignin removal. Oxygen delignification of primary clarifier sludge improved hydrolysis yield by up to 90%. However steam-exploded Douglas fir was very resistant to hydrolysis at low enzyme loading, and oxygen delignification decreased hydrolysis rate and yield. PMID- 11480920 TI - Immobilization and stabilization of invertase on Cajanus cajan lectin support. AB - Use of lectins as ligands for the immobilization and stabilization of glycoenzymes has immense application in enzyme research and industry. But their widespread use could be limited by the high cost of their production. In the present study preparation of a novel and inexpensive lectin support for use in the immobilization of glycoenzymes containing mannose or glucose residues in their carbohydrate moiety has been described. Cajanus cajan lectin (CCL) coupled covalently to cyanogen bromide activated Seralose 4B could readily bind enzymes such as invertase, glucoamylase and glucose oxidase. The immobilized and glutaraldehyde crosslinked preparations of invertase exhibited high resistance to inactivation upon exposure to enhanced temperature, pH, denaturants and proteolysis. Binding of invertase to CCL-Seralose was however found to be readily reversible in the presence of 1.0 M methyl alpha-D mannopyranoside. In a laboratory scale column reactor the CCL-Seralose bound invertase was stable for a month and retained more than 80% of its initial activity even after 60 days of storage at 4 degrees C. CCL-Seralose bound invertase exhibited marked stability towards temperature, pH changes and denaturants suggesting its potential to be used as an excellent support for the immobilization of other glycoenzymes as well. PMID- 11480921 TI - A new method for conservation of nitrogen in aerobic composting processes. AB - Several factors, such as pH, C/N ratio, temperature, mixing and turning, and aeration rate, could affect the loss of ammonia in composting reactions. Substantial loss of ammonia can reduce the nutrient value of the compost product and may lead to a severe odor problem in the composting facility. A new method for conservation of ammonia in composting was proposed and tested in this study. The ammonia being produced during the composting was precipitated into struvite crystals by addition of Mg and P salts. Ammonia volatilization was greatly reduced by this method and it also contributed to a remarkable increase in total ammoniacal-N (TAN) content in the compost, reaching up to 1.4% of dry mass. This value of TAN content was 3-5 times higher than that in normal compost. The scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) analyses confirmed the formation of struvite crystals in the aerobic composting process. PMID- 11480922 TI - Copper and zinc fractions affecting microorganisms in long-term sludge-amended soils. AB - The influences of Zn and Cu on soil enzyme activities (acid phosphatase, alkaline phosphatase, arylsulfatase, cellulase, dehydrogenase, protease (z-FLase), urease, beta-D-glucosidase and beta-D-fructofuranosidase (invertase)) and microbial biomass carbon were investigated in agricultural soils amended with municipal sewage sludge or compost since 1978. The trace metals in the soils were fractionated using a sequential extraction method. Long-term application of the sewage sludge and composts caused accumulations of Cu and Zn in the soils, ranging from 140 to 144 and from 216 to 292 mg kg(-1), respectively. The percentage of Cu was highest in the NaOH- and HNO3-extractable fractions (44-51% and 38-46%, respectively), while the percentage of Zn was highest in the HNO3- and EDTA-extractable fractions (65-83% and 11-32%, respectively). Although the percentage of the bioavailable fractions (sum of KNO3 + H2O-, NaOH-, and EDTA extractable amounts) of Cu (53-64%) was higher than that of Zn (15-37%), the percentage of the most labile fractions (KNO3 + H2O) of Zn (2.1-5.9%) was larger than that of Cu (1.1-2.4%). The size of the microbial biomass carbon increased with the application of sewage sludge or compost. For some enzymes, however, the ratio of the enzyme activity to microbial biomass was lower in the soils amended with sewage sludge or compost than that in the control soil. The soil enzyme activities were more adversely affected by Zn than by Cu. From a multiple regression analysis, it was found that dehydrogenase, urease, and beta-D glucosidase activities were reduced by the KNO3 + H2O-extractable fraction of Zn in the soils. These microbial activities seem to be sensitive to Zn stress, indicating the possibility that they might be useful bioindicators for evaluation of the toxic effects of Zn on microorganisms in the soils. PMID- 11480923 TI - Influence of one or two successive annual applications of organic fertilisers on the enzyme activity of a soil under barley cultivation. AB - The effect of cow manure and two rates of addition of municipal solid waste (MSW) compost on the enzymatic activity of a soil supporting barley cultivation was studied and compared with mineral fertilisation (MF). The experiment was carried out in unirrigated land in field conditions for two years. One set of plots was fertilised only once, at the start of the experiment, while another set of plots was fertilised annually (before each sowing). In general, the organic amendments stimulated soil enzyme activity but mineral fertilisation did not. The annual addition of large quantities of MSW compost, in general, led to lower levels of enzyme activity than similar rates of amendment in the plots receiving a single addition, probably because of the toxic effect which the heavy metals incorporated with the MSW compost had on microbial development. In the second year, plot receiving a single application of organic amendment showed higher protease hydrolysing casein, beta-glucosidase and dehydrogenase activities than control or soil with mineral fertilisation. This implied that the addition of organic waste, and particularly MSW compost, had a catalysing effect in the soil which lasted for the following years. Barley yields obtained with organic amendments were, in general, similar to, or even higher, than those obtained with mineral fertilisation. The annual addition of high doses of compost had an inhibitory effect on enzyme activity and barley yield (compared with the results obtained with the low addition of compost) due to the negative effect of the heavy metals incorporated with the compost. PMID- 11480924 TI - Generation of xylose solutions from Eucalyptus globulus wood by autohydrolysis posthydrolysis processes: posthydrolysis kinetics. AB - Eucalyptus wood samples were treated with water under selected operational conditions (autohydrolysis reaction) to obtain a liquid phase containing hemicellulose-decomposition products (mainly acetylated xylooligosaccharides, xylose and acetic acid). In a further acid-catalysed step (posthydrolysis reaction), xylooligosaccharides were converted into xylose, a carbon source for further fermentation. The kinetic pattern governing the posthydrolysis step was established by reacting xylooligosaccharide-containing liquors at 100.5 degrees C, 115 degrees C, 125 degrees C or 135 degrees C in media containing 0.5, 1.0, 1.5 or 2 wt% of catalyst (sulphuric acid). The time course of the concentrations of xylooligosaccharides, xylose, furfural and acetic acid were determined, and the results were interpreted by means of a kinetic model which allowed a close reproduction of the experimental data. Almost quantitative conversion of xylooligosaccharides into xylose was achieved under a variety of experimental conditions. The first-order, kinetic coefficient for xylooligosaccharide hydrolysis (k1, h(-1)) varied with both temperature (T, K) and molar sulphuric acid concentration (C) according to the equation In k1 = 36.66 + 1.00lnC - 108.0/(8.314T). The hydrolysis of acetyl groups followed a first-order kinetics. The corresponding kinetic coefficient (ka, h(-1) was correlated with the operational conditions by the equation Inka = 26.80+ 1.18 InC - 73.37/(8.314T). PMID- 11480925 TI - Nitrogen removal from wastewaters at low C/N ratios with ammonium and acetate as electron donors. AB - A denitrifying upflow anaerobic sludge blanket (UASB) reactor was operated at different nitrate loading rates at a C/N ratio of 1.2, with acetate as an electron donor. This resulted in an increase in the accumulation of nitrite. After this, the UASB reactor was supplemented with 100 mg NH4+-Nl(-1) d(-1), while acetate was gradually limited in the medium. This prevented nitrite accumulation at a C/N ratio of 0.6 due to an enhanced nitrite reduction rate achieved in the reactor. An increasing amount of ammonium was consumed when the C/N ratio was lowered in the medium. This suggested that ammonium was used as an alternative electron donor during denitrification, which is supported by nitrogen balances. Nitrite was shown to be toxic for the nitrogen removal process at 200 400 mg NO2--N(l(-1) when the C/N ratio was decreased to 0.4 leading to formation of ammonium. The present study showed that addition of ammonium as an alternative electron donor for denitrification achieved a nitrogen removal process with negligible accumulation of undesirable intermediates. PMID- 11480926 TI - Decolorization of an anthraquinone-type dye using a laccase formulation. AB - Decolorization of the dye Remazol Brilliant Blue R (RBBR) was studied, as it is representative of an important class of recalcitrant anthraquinone-type dyes. For this purpose a commercial laccase formulation (CLF) containing laccase, a redox mediator and a non-ionic surfactant was used. Small molecular weight components were removed from the CLF by gel filtration, which made it possible to compare the effect of its laccase alone. Apart from slightly better thermostability of the CLF as compared with the laccase alone, the pH and temperature profiles were similar regardless of the presence of the small molecular weight components. The laccase alone did not decolorize RBBR. A small molecular weight redox mediator (HBT) was necessary for decolorization to occur. A comparison of the kinetics of RBBR decolorization using the CLF and its laccase alone is reported. Provided that a redox mediator is included, it is suggested that laccase may be suitable for the wastewater treatment of similar anthraquinone dyes. PMID- 11480927 TI - Efficient use of animal manure on cropland--economic analysis. AB - Manure contains all the macro- and microelements needed for plant growth; however, it represents one of the most underutilized resources in the US. The major problem with the use of manure on cropland is the direct effect of its composition on application cost. This cost is a function of the mineralization process of organic matter. The mineralization process is influenced by the properties of the manure, properties of the soil, moisture, and temperature. This study evaluates the simultaneous effect of these variables on the optimal use of manure on cropland. The results show that the properties of manure and soil significantly affect the mineralization of organic nitrogen and thus the optimal quantity of manure required to satisfy the nutrient requirement of crops in a given rotation system. Manure application costs range from a low of 18% of the cost of commercial fertilizer for chicken manure applied to one type of soil, to a high of 125% of the cost of commercial fertilizer for cow manure applied to another type of soil. The maximum distance to transfer manure to the field, that will equate its application cost to the cost of commercial fertilizer, ranges from a high of 35 km (22 miles) for chicken manure applied to one type of soil, to a low of 1 km (0.62 miles) for cow manure applied to another type of soil. For rotation system 2, manure application costs range from a low of 37% of the cost of commercial fertilizer for chicken manure applied to one type of soil, to a high of 136% of the cost of commercial fertilizer for cow manure applied to another type of soil. The maximum distance to transfer manure to the field, that will equate its cost to the cost of commercial fertilizer, ranges from a high of 20 km (12.5 miles) for chicken manure applied to one type of soil, to a low of 0 km (0 miles) for cow manure applied to another type of soil. PMID- 11480928 TI - Assessing modulus of elasticity of wood-fiber cement (WFC) sheets using nondestructive evaluation (NDE). AB - This study evaluates whether the mechanical properties of modulus of rupture (MOR) and modulus of elasticity (MOE) of wood-fiber cement (WFC) sheets are correlated with the nondestructive parameters of stress wave velocity and density of the material. Longitudinal stress wave technique was used to evaluate WFC nondestructively using a total of 117 specimens (measuring each 241 x 51 mm) obtained from 39 WFC sheets. The aim was to establish the correlation between dynamic versus static MOE of the material for predicting the actual mechanical property. Even though short dimension specimens were used, results obtained were encouraging. A correlation coefficient (R) of 0.828 was found when the static MOE of the material was used as a function of nondestructive parameters. A multivariate linear regression analysis using the specimen's density, wave velocity, and dynamic MOE provided the strongest correlation to the static MOE. The correlation observed for MOR as a function of static MOE is within the normal range obtained for wood composites. A nondestructive evaluation (NDE) using full size WFC sheets is recommended and can probably improve the relationship between the static and the predicted MOE. PMID- 11480929 TI - Study of composting of cotton residues. AB - A study on the composting of residues of cotton gin previously crushed or uncrushed is presented in this work. A multifactor analysis of correlation for each one of the treatments revealed that the final results obtained during the composting process of cotton residuals were very different according to whether the residues were crushed or uncrushed. Generally, the highest final values of macronutrients and micronutrients obtained were found when the composting process was carried out with the crushed residuals. PMID- 11480930 TI - Effect of nutrients supplementation on anaerobic sludge development and activity for treating distillery effluent. AB - Startup of laboratory anaerobic reactors and treatment efficiency were investigated by supplementing the distillery effluent feed with macronutrients (Ca, P) and micronutrients (Ni, Fe and Co) under mesophilic conditions. Calcium and phosphate were deterimental to the treatment efficiency and sludge granulation. Traces of salts of iron, nickel and cobalt, individually and in combinations improved the COD removal efficiency and sludge granulation process. PMID- 11480931 TI - When half a word is enough: infants can recognize spoken words using partial phonetic information. AB - Adults process speech incrementally, rapidly identifying spoken words on the basis of initial phonetic information sufficient to distinguish them from alternatives. In this study, infants in the second year also made use of word initial information to understand fluent speech. The time course of comprehension was examined by tracking infants' eye movements as they looked at pictures in response to familiar spoken words, presented both as whole words in intact form and as partial words in which only the first 300 ms of the word was heard. In Experiment 1, 21-month-old infants (N = 32) recognized partial words as quickly and reliably as they recognized whole words; in Experiment 2, these findings were replicated with 18-month-old infants (N = 32). Combining the data from both experiments, efficiency in spoken word recognition was examined in relation to level of lexical development. Infants with more than 100 words in their productive vocabulary were more accurate in identifying familiar words than were infants with less than 60 words. Grouped by response speed, infants with faster mean reaction times were more accurate in word recognition and also had larger productive vocabularies than infants with slower response latencies. These results show that infants in the second year are capable of incremental speech processing even before entering the vocabulary spurt, and that lexical growth is associated with increased speed and efficiency in understanding spoken language. PMID- 11480932 TI - Mother-child conversational interactions as events unfold: linkages to subsequent remembering. AB - The study reported here was designed to examine linkages between mother-child conversational interactions during events and children's subsequent recall of these activities. In this longitudinal investigation, 21 mother-child dyads were observed while they engaged in specially constructed activities when the children were 30, 36, and 42 months of age. Analyses of the children's 1-day and 3-week recall of these events indicated that at all age points, features of the activities that were jointly handled and jointly discussed by the mother and child were better remembered than were features that were either (1) jointly handled and talked about only by the mother, or (2) jointly handled and not discussed. Potential linkages were also explored between incidental memory for personal experiences and deliberate recall of familiar but arbitrary materials. In this regard, children's recall of the special activities was positively correlated with their recall of objects in a deliberate memory task performed at 42 months. PMID- 11480933 TI - Individual differences in inhibitory control and children's theory of mind. AB - This research examined the relation between individual differences in inhibitory control (IC; a central component of executive functioning) and theory-of-mind (ToM) performance in preschool-age children. Across two sessions, 3- and 4-year old children (N = 107) were given multitask batteries measuring IC and ToM. Inhibitory control was strongly related to ToM, r = .66, p < .001. This relation remained significant controlling for age, gender, verbal ability, motor sequencing, family size, and performance on pretend-action and mental state control tasks. Inhibitory tasks requiring a novel response in the face of a conflicting prepotent response (Conflict scale) and those requiring the delay of a prepotent response (Delay scale) were significantly related to ToM. The Conflict scale, however, significantly predicted ToM performance over and above the Delay scale and control measures, whereas the Delay scale was not significant in a corresponding analysis. These findings suggest that IC may be a crucial enabling factor for ToM development, possibly affecting both the emergence and expression of mental state knowledge. The implications of the findings for a variety of executive accounts of ToM are discussed. PMID- 11480934 TI - Learning words from knowledgeable versus ignorant speakers: links between preschoolers' theory of mind and semantic development. AB - Two studies addressed whether children consider speakers' knowledge states when establishing initial word-referent links. In Study 1, forty-eight 3- and 4-year olds were taught two novel words by a speaker who expressed either knowledge or ignorance about the words' referents. Children showed better word learning when the speaker was knowledgeable. In Study 2, forty-eight 3- and 4-year-olds were taught two novel words by a speaker who expressed uncertainty about their referents. Whether the uncertainty truly reflected ignorance, however, differed across conditions. In one condition, the speaker said he made the object himself and thus, he was knowledgeable. In the other condition, the speaker stated that the object was made by a friend and thus, expressed ignorance about it. Four-year olds learned better in the speaker-made than in the friend-made condition; 3-year olds, however, showed relatively poor learning in both conditions. These findings suggest that theory-of-mind developments impact word learning. PMID- 11480935 TI - Prior knowledge and exemplar encoding in children's concept acquisition. AB - Three experiments examined how children's domain knowledge and observation of exemplars interact during concept acquisition and how exposure to novel exemplars causes revision of such knowledge. In Experiments 1 (N = 126) and 2 (N = 64), children aged 4 to 10 years were shown exemplars of fictitious animal categories that were either unrelated to, or consistent with, their prior knowledge in 25% or 75% of presented exemplars. In Experiment 3, children (N = 290) saw fictitious animal, artifact, or unfamiliar social categories that were either consistent or inconsistent with their prior knowledge in 20%, 40%, 60%, or 80% of exemplars. In the test, children made judgments about the likely co-occurence of features. In all experiments, prior knowledge and exemplar observation independently influenced children's categorization judgments. Utilization of prior knowledge was consistent across age and domain, but 10-year-olds were more sensitive to observed feature covariation. Training with larger categories increased the impact of observed feature covariation and decreased reliance on prior knowledge. PMID- 11480936 TI - The development of self-regulation in the first four years of life. AB - This study examined longitudinally the development of self-regulation in 108 young children during the first 4 years of life. Children's committed compliance (when they eagerly embraced maternal agenda) and situational compliance (when they cooperated, but without a sincere commitment) were studied. Both forms of compliance were observed in "Do" contexts, in which the mothers requested that the children sustain unpleasant, tedious behavior, and in "Don't" contexts, in which they requested that the children suppress pleasant, attractive behavior. Children's internalization while alone in the similar contexts was also studied. Parallel assessments were conducted when the children were 14, 22, 33, and 45 months of age. At all ages, the Do context was much more challenging for children than the Don't context. Girls surpassed boys in committed compliance. Both forms of compliance were longitudinally stable, but only within a given context. Children's fearfulness and effortful control, observed and mother reported, correlated positively with committed compliance, but mostly in the Don't context. Committed, but not situational, compliance was linked to children's internalization of maternal rules, observed when the children were alone in the Do and Don't contexts. These links were both concurrent and longitudinal, context specific, and significant even after controlling for maternal power assertion. There was modest preliminary evidence that committed compliance may generalize to interactions with adults other than the mother. PMID- 11480937 TI - The relations of regulation and emotionality to children's externalizing and internalizing problem behavior. AB - The purpose of this study was to examine the relation of different types of negative emotion and regulation and control to 55- to 97-month-olds' internalizing and externalizing problem behaviors. Parents and teachers provided information on children's (N = 214) adjustment, dispositional regulation and control, and emotion, and children's regulation was observed during several behavioral tasks. Internalizing was defined in two ways: as social withdrawal (to avoid overlap of items with measures of emotionality) or, more broadly, as anxiety, depression, and psychosomatic complaints. In general, children with externalizing problems, compared with children with internalizing problems and nondisordered children, were more prone to anger, impulsivity, and low regulation. Children with internalizing symptoms were prone to sadness, low attentional regulation, and low impulsivity. Relations between internalizing problems and emotionality were more frequent when the entire internalizing scale was used. Findings suggest that emotion and regulation are associated with adjustment in systematic ways and that there is an important difference between effortful control and less voluntary modes of control. PMID- 11480938 TI - The peer group as a context for the development of young adolescent motivation and achievement. AB - This study investigated the peer group as a context for the socialization of young adolescents' motivation and achievement in school. Social network analysis was used to identify peer groups of adolescents in middle school whose members regularly interacted with each other (N = 331). Actual reports from these peer group members were used to assess peer group characteristics. Multilevel analyses indicated that peer groups did socialize some academic characteristics, controlling for selection factors. Students' peer group context in the fall predicted changes in their liking and enjoyment of school (intrinsic value) and their achievement over the school year. Students' peer group context was unrelated to changes in their beliefs about the importance of school (utility value) or expectancies for success over the school year. PMID- 11480939 TI - When groups are not created equal: effects of group status on the formation of intergroup attitudes in children. AB - This study was designed to examine whether the presence of implicit links between social groups and high versus low status attributes affects the formation of intergroup attitudes. Elementary school children aged 7 to 12 years (N = 91) were given measures of classification skill and self-esteem, and assigned to one of three types of summer school classrooms in which teachers made (1) functional use of novel ("blue" and "yellow") social groups that were depicted via posters as varying in status, (2) no explicit use of novel social groups that were, nonetheless, depicted as varying in status, or (3) functional use of novel social groups in the absence of information about status. After 6 weeks, children completed measures of intergroup attitudes. Results indicated that children's intergroup attitudes were affected by the status manipulation when teachers made functional use of the novel groups. Children who were members of high-status (but not low-status) groups developed in-group biased attitudes. PMID- 11480940 TI - Interparental discord and adolescent adjustment trajectories: the potentiating and protective role of intrapersonal attributes. AB - This prospective study of 360 adolescent-mother dyads examined whether associations between marital discord and trajectories of adolescent depressive symptoms and delinquency varied as a function of three intrapersonal attributes: temperament, childhood behavior problems, and perceived family support. Difficult temperament (i.e., dysrhythmicity, poor task orientation) potentiated the effects of marital discord on adolescent trajectories of adjustment, whereas heightened perceptions of family support protected adolescents from the adverse effects of marital discord. Adolescents with behavior problem histories were initially less vulnerable to marital discord; however, the high levels of depressive symptoms exhibited by adolescents with childhood behavior problems persisted over time only when they were exposed to elevated marital discord. The effects of the moderators differed in terms of duration and course. PMID- 11480941 TI - Interpersonal influence in family systems: a social relations model analysis. AB - Is parental control of a child's behavior due to the parenting style of the parent (e.g., authoritative parenting), the child's inclination toward compliance (i.e., an "easy" child), or the reciprocal system of parent-child exchange? This study addressed this question, as well as the broader one of who drives the interaction whenever one family member influences another. Family members from 208 two-parent two-child families of adolescents rated the degree to which each person in the family influenced each of the other family members. Social relations analysis of these data indicated that characteristics of the partner and the unique actor-partner "fit" were systematic sources of interpersonal influence, and that reciprocal influence was present in most--but not all--family dyads. A pattern in which parents compensate for each other's influenceability was also found. It is recommended that these findings be replicated using observational data that have been sequentially analyzed. PMID- 11480942 TI - Young children's feelings about school. AB - This study examined factors associated with young children's feelings about school in kindergarten and first grade, using a new measure, the Feelings about School (FAS). The FAS measures children's perceptions of academic competence, their feelings about the teacher, and their general attitudes toward school. Findings provided support for the reliability and validity of the FAS for kindergartners (N = 225) and first graders (N = 127). Variables presumed to predict children's feelings about school were the classroom structure, academic performance, and relationships with teachers. Feelings about school were expected to predict academic engagement. Correlational analyses indicated that kindergartners' and first graders' feelings about school were associated with their academic skills, as measured by direct assessments and teacher ratings. The evidence for first graders was stronger than for kindergartners. Kindergartners' general attitudes toward school were more negative in highly structured, teacher directed classroom environments. First graders' perceptions of competence were more negative in classrooms lacking structure and control. First graders', but not kindergartners', perceptions of competence were significantly associated with academic engagement. PMID- 11480943 TI - Children and mothers in war: an outcome study of a psychosocial intervention program. AB - The present study was designed to evaluate the effects on children (age: M = 5.5 years) in war-torn Bosnia and Herzegovina of a psychosocial intervention program consisting of weekly group meetings for mothers for 5 months. An additional aim was to investigate the children's psychosocial functioning and the mental health of their mothers. Internally displaced mother-child dyads were randomly assigned to an intervention group receiving psychosocial support and basic medical care (n = 42) or to a control group receiving medical care only (n = 45). Participants took part in interviews and tests to provide information about war exposure, mental health, psychosocial functioning, intellectual abilities, and physical health. Results showed that although all participants were exposed to severe trauma, their manifestations of distress varied considerably. The intervention program had a positive effect on mothers' mental health, children's weight gain, and several measures of children's psychosocial functioning and mental health, whereas there was no difference between the two groups on other measures. The findings have implications for policy. PMID- 11480944 TI - The influence of neighborhood disadvantage, collective socialization, and parenting on African American children's affiliation with deviant peers. AB - This study focused on hypotheses about the contributions of neighborhood disadvantage, collective socialization, and parenting to African American children's affiliation with deviant peers. A total of 867 families living in Georgia and Iowa, each with a 10- to 12-year-old child, participated. Unique contributions to deviant peer affiliation were examined using a hierarchical linear model. Community disadvantage derived from census data had a significant positive effect on deviant peer affiliations. Nurturant/involved parenting and collective socialization processes were inversely associated, and harsh/inconsistent parenting was positively associated, with deviant peer affiliations. The effects of nurturant/involved parenting and collective socialization were most pronounced for children residing in the most disadvantaged neighborhoods. PMID- 11480945 TI - Parents' roles in shaping early adolescents' occupational aspirations. AB - Relations among dimensions of parenting and adolescents' occupational aspirations were examined in two specific domains: academics and sports. The sample consisted of 444 seventh graders, with approximately equal numbers of African American and European American males and females, from two-parent nondivorced families. Multiple measures were used as indicators of parents' values and behaviors, youths' values and beliefs, positive identification with parents, and adolescents' occupational aspirations. In the academic domain, parents' values predicted youths' values directly rather than indirectly through their behaviors. In contrast, fathers' behaviors mediated the relation between parents' and youths' values in the sports domain. Positive identification was directly related to adolescents' values (especially about academics); however, positive identification did not moderate the transmission of values from parent to child in either domain. Parents' values predicted adolescents' occupational aspirations via both direct and indirect pathways. Similar results were obtained for African American and European American males and females. These findings highlight the potential role of parents as socializers of achievement-related values, and, ultimately, adolescents' occupational visions of themselves in the future. PMID- 11480946 TI - Adolescent, parent, and observer perceptions of parenting: genetic and environmental influences on shared and distinct perceptions. AB - Agreement among reporters on features of family life, whether family members or outside observers, is considered to be low. This study, which involved a national sample of 720 families comprised of identical and fraternal twins, full siblings, half siblings, and biologically unrelated stepsiblings, examined the issue of low interrater agreement by decomposing the common and unique variance among parent, child, and observer reports of parenting behaviors (warmth and negativity) into genetic and environmental factors. Quantitative genetic analyses were employed to decompose the "Social" level of perception (common variance among parents, children, observers), the level of "Family" subculture (common variance only among parents and children), and the unique "Individual" level into genetic and environmental components. It was predicted that genetic factors would account for substantial portions of the variance at the Social and Family levels; nonshared environmental factors were expected to influence variance unique to child reports; and shared environmental factors were expected to influence variance unique to parent reports. A second and related aim of the study was to examine the subjective-objective dimension of genetic effects on measures of the environment. Results of model-fitting analyses generally supported the predictions for parental warmth and negativity at the Family and Individual levels. At the Social level, genetic factors were predominant for parental negativity and shared environmental factors for parental warmth. The findings are discussed in terms of genetically influenced child effects on parenting and methodological difficulties in constructing latent variables. PMID- 11480947 TI - Visual-proprioceptive intermodal perception using point light displays. AB - Three experiments explored 5- and 7-month-old infants' intermodal coordination of proprioceptive information produced by leg movements, and visual movement information specifying these same motions. The visual information took the form of point light information for leg and feet movements, with visual displays presented in upright, ego-centered on-joint (Experiment 1, N = 48); upright, ego centered off-joint (Experiment 2, N = 48); and inverted, observer-centered off joint (Experiment 3, N = 48) orientations. Measures of preferential looking indicated intermodal perception in infants of both ages while seeing on-joint, ego-centered orientations, and for 7-month-olds (and possibly 5-month-olds) while seeing off-joint, ego-centered displays; neither age group demonstrated intermodal perception for off-joint, observer-centered displays. These results suggest that coordination of visual and proprioceptive inputs is constrained by infants' information processing of the displays, and have implications for infants' growing understanding of their self-movement and the development of knowledge of the self. PMID- 11480948 TI - Extended visual fixation and distractibility in children from six to twenty-four months of age. AB - Distractibility during extended visual fixations in children 6 months to 2 years of age was examined. A children's Sesame Street movie (Follow That Bird) was presented to children (N = 40) for a minimum of 20 min while fixation was videotaped and heart rate was recorded. Distractors (computer-generated patterns or another Sesame Street movie) were presented on an adjacent television screen. Consistent with prior research with older preschool-age children, the latency to turn toward the distractor was a function of the length of the look occurring before distractor onset. For the period immediately before distractor onset, children had a greater sustained lowered heart rate for the trials on which they continued looking at the center television monitor than for the trials on which they looked toward the distractor. This pattern of distractibility suggests attention increases over the course of a look toward the television, and that heart rate changes reflect this increase in attention. PMID- 11480949 TI - How infants use vision for grasping objects. AB - The role of vision was examined as infants prepared to grasp horizontally and vertically oriented rods. Hand orientation was measured prior to contact to determine if infants differentially oriented their hands relative to the object's orientation. Infants reached for rods under different lighting conditions. Three experiments are reported in which (1) sight of the hand was removed (N = 12), (2) sight of the object was removed near the end of the reach (N = 40, including 10 adults), and (3) sight of the object was removed prior to reach onset (N = 9). Infants differentially oriented their hand to a similar extent regardless of lighting condition and similar to control conditions in which they could see the rod and hand throughout the reach. In preparation for reaching, infants may use the current sight of the object's orientation, or the memory of it, to orient the hand for grasping; sight of the hand had no effect on hand orientation. PMID- 11480950 TI - Planar asymmetry tips the phonological playground and environment raises the bar. AB - Reading readiness varies as a function of family and environmental variables. This study of 11-year-old children (N = 39) was designed to determine if there was an additional or interactive contribution of brain structure. Evidence is presented that both environmental and biological variables predict phonological development. Temporal lobe (planar) asymmetry, hand preference, family history of reading disability, and SES explained over half of the variance in phonological and verbal performance. The results demonstrate a linear association between cerebral organization and phonological skill within socioeconomic groups. These data provide concrete evidence to support the commonly held assumption that both environmental and biological factors are independent determinants of a child's ability to process linguistic information. PMID- 11480951 TI - Notions for mental health: a WHO public health initiative. PMID- 11480952 TI - Prominence of symptoms and level of stigma among depressed patients in Calcutta. AB - A pilot study of 29 patients for research on clinical depression employed the framework of cultural epidemiology to examine illness-related experience, meaning, behaviour with a Bengali version of the explanatory Model Interview Catalogue (EMIC). This report examined patterns of distress and stigma with reference to the most troubling patient-specified symptom. All subjects were psychiatric outpatients in the Institute of Psychiatry, Calcutta, and met Diagnostic and Statistical Manual (DSM)-IV criteria for a major depressive episode. Only 5 patients (17.2%) identified sadness as the most troubling problem, and 48.3% specified pains and other somatic symptoms instead. An internally consistent stigma scale with Cronbach's alpha of .67, comprised 13 items, was used to assess stigma. In addition to the scale score for each subject, the contribution of each item was reported and compared. A suggestive, though not significantly lower value of the stigma score for patients reporting somatic symptoms as most troubling, compared with sadness, was consistent with findings from prior studies showing a positive relationship between the magnitude of depression and stigma. These findings are discussed with reference to their impact on recognition and help seeking among patients, and recognition and management of depression by general practitioner. Culturally distinctive presentations and social contexts of depression and other mental illnesses should be addressed in professional training and public health communications. PMID- 11480953 TI - Psychiatry in non-psychiatric setting--a comparative study of physicians and surgeons. AB - The paper reports awareness of a group of physicians and surgeons about psychiatric disorders in clinical practice and their attitudes towards psychiatric referral. The study was carried out in a teaching general hospital. Sample consisted of 60 faculty members and senior residents belonging to various clinical disciplines. A 14-item structured questionnaire was used for assessment. The data were compared across the specialists from medical and surgical specialities. The main findings included underestimation of psychiatric morbidity by all the clinicians and a general reluctance to refer patients to a psychiatrist for varied reasons. However, most doctors considered psychiatric referral as useful. No major differences were noticed between surgeons and physicians regarding their attitudes and awareness about psychiatry. There is a need to increase the awareness of non-psychiatric clinicians about psychiatric disorders and to develop consultation-liaison psychiatric units and expand undergraduate medical education programmes in psychiatry in the developing countries. PMID- 11480954 TI - Depression in medical disorders. AB - This review highlights the occurrence of depressive disorders in medically ill patients. It discusses the difficulties in diagnosing depression in the medically ill due to the vegetative symptoms of depression which are also a feature of many medical diseases. Depression in the medically ill may be due to the disease itself, its psychological reaction, due to complications because of metabolic or other disturbances, or due to the medications for the treatment of the disease. The role of active consultation liaison between medical specialists and mental health professionals will help provide total care to the medically ill. PMID- 11480955 TI - Depression in children and adolescents. AB - Childhood depression accounts for a substantial proportion of cases in child psychiatric clinics. Depression is more common in boys than girls in school age children and it becomes reverse during adolescents. The childhood depression can be wholly chemical, wholly due to psychological factors of a combination of both. The causes of depression can be discussed under three headings--genetic factors, biological factors and social factors. Some children display the classical symptoms of sadness, anxiety, restlessness, eating and sleeping problems while others present with physical problems like aches and pains which do not respond to treatment. Diagnosis depends on interview or questionnaire instruments to screen populations and some biological procedures to determine neuro-endocrine and other physical dysfunctions. Treatment consists of pharmacological treatment and psychological treatment. Tricyclic antidepressants, serotonin specific reuptake inhibitors and electroconvulsive therapy constitute pharmacological part of the treatment, whereas supportive, insight oriented, play, behaviour, family therapies and cognitive psychotherapy constitute psychological part. PMID- 11480956 TI - Depressive disorders in old age. AB - People are living longer life since with the improvement in health and medical services provided. Developing countries are not lagging behind the developed world as far as the life expectancy is concerned. The principal mental disorders of elderly people are mood disorders and dementia. In older people living in a community the rates of depressive disorders are likely to be the same as for all age groups of the general population, yet its prevalence in old age institution is high. Aetiology of depression in old age is caused by a variety of conditions namely genetic factors, physiological changes in all the systems, disability, loss of resources, some medical illness, etc. Clinical features include depressed mood most of the day, significant weight loss, persistent sleeplessness, fatigue, feelings of worthlessness, recurrent thoughts of death and suicidal attempt, etc. Tricyclic antidepressants are effective in elderly depressives. Selective serotonin re-uptake inhibitors are well tolerated by elderly patients and are effective too. Electroconvulsive therapy is an alternative treatment of acute depression. Psychotherapy is often helpful and long-term care should be planned for to reduce disability. PMID- 11480957 TI - Management of chronic stable angina pectoris from a sceptic's view point. AB - Ischaemic heart disease is a leading cause of death in the world. It has clinically defined phases as: Asymptomatic, stable angina, progressive angina and unstable angina. It is important to differentiate patients of angina into those with stable and unstable angina--risk stratification and management differ in the two groups. Risk stratification of patients with stable angina using clinical parameters helps in development of clearer indication of referral for exercise testing and cardiac catheterisation. Chronic stable angina patients with history of documented myocardial infarction of Q waves on ECG should have measurement of left ventricular systolic function (ie, ejection fraction) as it is important for choosing the appropriate medical or surgical therapy. Symptomatic patients with suspected or known coronary artery disease should usually undergo exercise testing to assess the risk of future cardiac events. The treatment of stable angina has two purposes: To prevent myocardial infarction and death and therapy directed towards preventing death. Pharmacotherapy consists of: Aspirin, lipid lowering agents, beta-blockers, nitrates, short acting dihydropyridine calcium antagonists, etc. For surgery, there are two well established approaches of revascularisation. One is coronary artery by-pass grafting and the other is percutaneous transluminal coronary angioplasty. Studies comparing different treatment modalities are elaborated in this article. In conclusion, it can be said that patients having severe symptoms affecting quality of life despite optimal medical therapy should be referred for revascularisation surgery. PMID- 11480958 TI - Selection of antidepressant drugs in general practice. AB - Depression is a common psychiatric illness and since majority of patients suffering from depression are first seen and treated by general practitioners, it is important for them to identify and treat depressive illness more effectively. Fortunately, depression is a treatable condition. Identifying the optimal antidepressant agent requires careful consideration of the patient's age, health status, and history of response to antidepressants. Other considerations include adverse effect profile, cost of drug therapy and convenient dosage schedule. Selective serotonin reuptake inhibitors are well tolerated and are considered by many to be the agents of choice in primary care treatment of depression because of their favourable adverse effect profile. Heterocyclic antidepressants are preferred in elderly patients and patients with medical problems. Tricyclic antidepressants can also be of great help specially to younger group of patients and patients who have failed to respond to other antidepressants. PMID- 11480959 TI - Ethnic differences in cancer mortality trends in the US, 1950-1992. AB - OBJECTIVE: To describe long-term mortality trends by ethnicity, sex, and age for selected cancers and to assess the effect of age-adjustment using different standard populations on rate ratios and rate differences comparing black to white cancer mortality. DESIGN: Mortality rates for selected cancers were obtained from published reports of the Vital Statistics of the United States (1950-1992). All ethnic- and sex-specific cancer rates were directly age-adjusted to the total 1970 US standard population and to a subset of the 1970 US standard population 40 years and older. RESULTS: Over a 42-year period, lung cancer mortality increased in all population subgroups. Colorectal cancer mortality declined in whites, but increased in blacks. Prostate cancer mortality increased slightly in white men, but dramatically increased in black men. Breast cancer mortality stabilized in white women, but increased markedly in black women. Uterine cancer mortality declined for both ethnicities, while ovarian cancer mortality rates increased for both ethnicities. As expected, the ratios of the age-adjusted cancer mortality rates comparing blacks to whites were the same regardless of the age structure used as the standard population. In contrast, the differences in the age-adjusted rates between blacks and whites were greater when the age-truncated standard population was used. CONCLUSIONS: There are unexplained ethnic differences in the long-term mortality trends of selected cancers. Of particular concern are the increasing death rates in black individuals from colorectal, prostate, breast, and ovarian cancers. Since almost all deaths from these cancers occur in persons over 40, age-adjustment using an age-truncated standard population that includes only those age groups at risk should be considered, particularly when the question to be addressed is one dealing with the impact of a characteristic, such as ethnicity or sex, on mortality risk. PMID- 11480960 TI - Longitudinal study of socio-economic differences in mortality among South Asian and West Indian migrants. AB - AIM: To examine socio-economic differences in mortality of South Asian and West Indian migrants. DESIGN: ONS Longitudinal Study, a 1% sample of the population of England and Wales. PARTICIPANTS: First generation South Asian and West Indian migrants present at the 1971 and 1981 censuses, aged under 65 years in 1981, were classified by occupational class, access to cars, housing, and a deprivation score. OUTCOME MEASURES: Relative risks of mortality, 1981-1997. RESULTS: There was considerable upward social mobility but most people remained socially stable, mainly relatively disadvantaged compared to all other study members. High levels of owner occupied housing among South Asians at both censuses were an exception. Socio-economic differentials in mortality were significant with car access in 1981 (relative risk 1.32, 95% confdence interval 1.03-1.70), and housing (rented vs owner occupied relative risk 1.50, 95% confidence interval 1.08-2.08) among South Asians and, among West Indians with car access in 1981 (relative risk 1.43, 95% confidence interval 1.05-1.94). Mortality was not significantly associated with socioeconomic circumstances in 1971. CONCLUSION: The disparity between those who remained advantaged or who moved up, and those who remained persistently disadvantaged, would have been a factor underlying the stronger differentials by later socio-economic circumstances. As these migrants age an increase in health inequalities could be expected. PMID- 11480961 TI - Psychological distress and coping in sickle cell disease: comparison of British and Jamaican attitudes. AB - OBJECTIVE: To investigate possible differences in coping mechanisms in the painful crisis between Jamaican and London patients with homozygous sickle cell disease. DESIGN: Patients recruited from two London hospitals and the MRC Laboratories (Jamaica) at the University of the West Indies, Kingston, Jamaica were assessed using a questionnaire design. Patients with homozygous sickle cell disease were included, 30 in London and 30 in Jamaica. RESULTS: Jamaican patients in Jamaica had less general anxiety, a lower emotional response to pain, lower levels of perceived pain, and felt better able to decrease their pain. London patients believed that the disease had a more marked effect on their quality of life. CONCLUSION: Understanding the differences between patients' response to pain and their coping ability between Jamaican and UK patients may have important lessons for evolving effective management in the UK. PMID- 11480962 TI - Assessment of dietary intake among Moroccan women and Surinam men. AB - OBJECTIVE: To establish a method for food consumption data collection in ethnic groups in the Netherlands. METHODS: Two pilot studies have been carried out, one among Moroccan women and one among Surinam men. First, focus group discussions were held to obtain background information from members of the target population. For food consumption assessment 2 X 24-h recall was chosen. Furthermore, background information on age, place of birth, frequency of visits to Morocco or Surinam, Dutch language skills, meal pattern, food purchases and antropometric measurements were taken. RESULTS: It was found that the diets of both Moroccan women and Surinam men tended to approach the recommended daily intake of macronutrients more than the average intake of a Dutch person. However, reported energy and micronutrient intake on the second day of the dietary recall was lower than on the first recall day. CONCLUSION: The 24-h recall is a suitable method for food consumption measurements among Moroccan and Surinam people. On the other hand, the interviewer must be aware of potential under-reporting, especially during the second recall. PMID- 11480963 TI - Immigration and leisure-time physical inactivity: a population-based study. AB - OBJECTIVE: To investigate the relationship between migration status and sedentary leisure-time physical activity status in the city of Malmo, Sweden. METHODS: The public health survey in 1994 is a cross-sectional study. A total of 5,600 individuals aged 20-80 completed a postal questionnaire. The response rate was 71%. The population was categorized according to country of birth. Multivariate analysis was performed using a logistic regression model to investigate the importance of possible confounders for the differences in sedentary leisure-time physical activity status. RESULTS: The prevalence of a sedentary leisure-time physical activity status was 18.1% among men and 26.7% among women. The odds ratio of a sedentary leisure-time physical activity status was significantly higher among men born in Arabic-speaking countries, in All other countries, and among women born in Yugoslavia, Poland, Arabic-speaking countries, and the category all other countries', compared to the reference group born in Sweden. The multivariate analysis including age, sex, and education did not alter these results. CONCLUSION: There were significant ethnic differences in leisure-time physical activity status. This is a CVD risk factor that could be affected by intervention programs aimed at specific ethnic subgroups of the population. PMID- 11480964 TI - Preventable risk factors for coronary heart disease and stroke amongst ethnic groups in London. AB - OBJECTIVES: People of African Caribbean descent have higher mortality rates from stroke than other ethnic groups. However, little is known about the prevalence of stroke risk factors in UK ethnic minority groups. We investigated the prevalence of these risk factors amongst African Caribbeans, black Africans and whites. DESIGN: A random sample of patients aged 45-74 registered with 16 general practices in south London was surveyed in 1995. Main outcome measures were: prevalence of hypertension, mean serum cholesterol, serum fibrinogen and glycosylated haemoglobin AIC. Logistic and linear regressions were used to determine ethnic differences in these measures. RESULTS: Hypertension was more prevalent in black Caribbeans (79.4%) and black Africans (71.6%) than in whites (54.3%) (p < 0.0001). There were ethnic group differences in mean random total cholesterol (p < 0.0001), triglycerides (p < 0.0001), fibrinogen (p = 0.03), HDL (p = 0.02) and HbAIC (p < 0.0001). Whites had higher mean random total cholesterol, triglycerides and fibrinogen than black Caribbeans but lower HDL and HbA1C. Black Africans had similar rates to black Caribbeans for these risk factors apart from lower triglvceride levels. CONCLUSION: These differences in risk factors may partially explain the high stroke mortality rates in black Caribbeans and black Africans compared to whites. There was little difference in prevalence of these risk factors between black Caribbean and black African groups. Specific strategies targeted to each ethnic group need to be developed to reduce risk factors. PMID- 11480965 TI - Ethnic differences in behavioural risk factors for stroke: implications for health promotion. AB - OBJECTIVES: Ethnic minority groups are at a higher risk of stroke and heart disease. However, designing effective prevention strategies requires responding to the needs of different ethnic groups. The aims of this study were to estimate the prevalence of four behavioural risk factors (smoking, drinking, exercise and weight) for stroke among Black Caribbeans, Black Africans and Whites, and also to examined reported willingness to change these behaviours. DESIGN: A random sample of 311 Black Caribbean, 300 White, and 105 Black Africans aged 45-74 registered with 16 practices in south London were surveyed in 1995. Information was obtained on smoking, drinking and exercise patterns, body mass index and perceptions of being at risk of stroke, and willingness to change risk behaviour. RESULTS: White respondents (31% age and sex standardised prevalence) were more likely to smoke than Black Caribbeans (23%) and Black Africans (10%) (p < 0.001). Self reported rates of drinking were higher than the government's 'sensible drinking levels' for 19% of Whites, 11% of Black Caribbeans and 4% Black Africans (p < 0.001). In contrast, fewer Whites (51%) were overweight (BMI > 27) than Black Caribbeans (60%) and Black Africans (68%) (p = 0.001). A high proportion of smokers wished to give up (89% Black African; 83% Black Caribbean; 74% White). A higher proportion of Black Caribbeans (35%) reported a willingness to reduce their alcohol intake compared to only 15% of Whites (p = 0.040). There was a difference between groups in attitudes to weight reduction with 69% Black Caribbean women expressing a desire to be thinner compared to 86% Whites and 82% Black Africans (p = 0.051). CONCLUSION: Strategies to reduce behavioural risk factors for heart attack and stroke need to emphasise different risk foctors among ethnic groups, especially in relation to alcohol use in the White population and weight in the Black Caribbean population. Influencing the change of these behaviours requires working in partnership with local community' groups. PMID- 11480966 TI - Experimental and theoretical studies on the thermal decomposition of heterocyclic nitrosimines. AB - A series of substituted 2-nitrosiminobenzothiazolines (2) were synthesized by the nitrosation of the corresponding 2-iminobenzothiazolines (6). Thermal decomposition of 2a--f and of the seleno analogue 7 in methanol and of 3-methyl-2 nitrosobenzothiazoline (2a) in acetonitrile, 1,4-dioxane, and cyclohexane followed first-order kinetics. The activation parameters for thermal deazetization of 2a were measured in cyclohexane (Delta H(++) = 25.3 +/- 0.5 kcal/mol, Delta S(++) = 1.3 +/- 1.5 eu) and in methanol (Delta H(++) = 22.5 +/- 0.7 kcal/mol, Delta S(++) = -12.9 +/- 2.1 eu). These results indicate a unimolecular decomposition and are consistent with a proposed stepwise mechanism involving cyclization of the nitrosimine followed by loss of N(2). The ground state conformations of the parent nitrosiminothiazoline (9a) and transition states for rotation around the exocyclic C==N bond, electrocyclic ring closure, and loss of N(2) were calculated using ab initio molecular orbital theory at the MP2/6-31G* level. The calculated gas-phase barrier height for the loss of N(2) from 9a (25.2 kcal/mol, MP4(SDQ, FC)/6-31G*//MP2/6-31G* + ZPE) compares favorably with the experimental barrier for 2a of 25.3 kcal/mol in cyclohexane. The potential energy surface is unusual; the rotational transition state 9a-rot-ts connects directly to the orthogonal transition state for ring-closure 9aTS. The decoupling of rotational and pseudopericyclic bond-forming transition states is contrasted with the single pericyclic transition state (15TS) for the electrocyclic ring-opening of oxetene (15) to acrolein (16). For comparison, the calculated homolytic strength of the N--NO bond is 40.0 kcal/mol (MP4(SDQ, FC)/6 31G*//MP2/6-31G* + ZPE). PMID- 11480967 TI - Influence of remote substituents on the equatorial/axial selectivity in the monooxygenation of methylene C--H bonds of substituted cyclohexanes. AB - The reactivity of individual C--H bonds in the methyl(trifluoromethyl)dioxirane TFDO oxygenation of stereogenic methylene groups in conformationally homogeneous monosubstituted cyclohexanes (2) has been determined. The unexpectedly high occurrence of O-atom insertion into C--H(ax) bonds suggests an in plane trajectory attack in the oxygenation while the diastereoselectivity of the reaction is qualitatively interpreted on the basis of the distinct hyperconjugative stabilization by the substituent of diastereomeric transition states due to long-range through bond interactions. PMID- 11480968 TI - Primary kinetic isotope effects on hydride transfer from 1,3-dimethyl-2 phenylbenzimidazoline to NAD(+) analogues. AB - Primary kinetic isotope effects (KIE) have been determined spectrophotometrically for the reaction of NAD(+) analogues (pyridinium, quinolinium, phenanthridinium, and acridinium ions) with 1,3-dimethyl-2-phenylbenzimidazoline in a 4:1 mixture of 2-propanol and water by volume at 25 degrees C. The values of KIE varied systematically from 6.27 to 4.06 as the equilibrium constant changed from around 10 to around 10(12). This is consistent with Marcus theory of atom transfer, assuming that there are no high-energy intermediates. Within this theory, the perpendicular effect is responsible for most of the change in KIE. The Marcus theory of atom transfer is consistent with a linear, triatomic model of the reaction. Perpendicular effects arise from the systematic decrease of bond distances and increase of bond orders in the critical complexes of the two related degenerate hydride transfer reactions as their C--H bonds become stronger. The parallel effect (Leffler--Hammond effect) is attenuated by the fairly high intrinsic barrier (lambda/4 is around 92 kJ/mol) and makes a smaller contribution to the change in the KIE. PMID- 11480969 TI - Atom transfer radical polymerization from nanoparticles: a tool for the preparation of well-defined hybrid nanostructures and for understanding the chemistry of controlled/"living" radical polymerizations from surfaces. AB - Structurally well-defined polymer--nanoparticle hybrids were prepared by modifying the surface of silica nanoparticles with initiators for atom transfer radical polymerization and by using these initiator-modified nanoparticles as macroinitiators. Well-defined polymer chains were grown from the nanoparticle surfaces to yield individual particles composed of a silica core and a well defined, densely grafted outer polystyrene or poly(methyl methacrylate) layer. In both cases, linear kinetic plots, linear plots of molecular weight (M(n)) versus conversion, increases in hydrodynamic diameter with increasing conversion, and narrow molecular weight distributions (M(w)/M(n)) for the grafted polymer samples were observed. Polymerizations of styrene from smaller (75-nm-diameter) silica nanoparticles exhibited good molecular weight control, while polymerizations of methyl methacrylate (MMA) from the same nanoparticles exhibited good molecular weight control only when a small amount of free initiator was added to the polymerization solution. The difference in polymerization behavior for styrene and MMA was ascribed to the facts that styrene undergoes thermal self-initiation while MMA does not and that termination processes involving freely diffusing chains are faster than those involving surface-bound chains. The polymerizations of both styrene and MMA from larger (300-nm-diameter) silica nanoparticles did not exhibit molecular weight control. This lack of control was ascribed to the very high initial monomer-to-initiator ratio in these polymerizations. Molecular weight control was induced by the addition of a small amount of free initiator to the polymerization but was not induced when 5--15 mol % of deactivator (Cu(II) complex) was added. PMID- 11480970 TI - Amino acid nitrosation products as alkylating agents. AB - Nitrosation reactions of alpha-, beta-, and gamma-amino acids whose reaction products can act as alkylating agents of DNA were investigated. To approach in vivo conditions for the two-step mechanism (nitrosation and alkylation), nitrosation reactions were carried out in aqueous acid conditions (mimicking the conditions of the stomach lumen) while the alkylating potential of the nitrosation products was investigated at neutral pH, as in the stomach lining cells into which such products can diffuse. These conclusions were drawn: (i) The alkylating species resulting from the nitrosation of amino acids with an -NH(2) group are the corresponding lactones; (ii) the sequence of alkylating power is: alpha-lactones > beta-lactones > gamma-lactones, coming respectively from the nitrosation of alpha-, beta-, and gamma-amino acids; and (iii) the results obtained may be useful in predicting the mutagenic effectiveness of the nitrosation products of amino acids. PMID- 11480971 TI - Tunable swelling kinetics in core--shell hydrogel nanoparticles. AB - Thermoresponsive, core--shell poly-N-isopropylacrylamide (p-NIPAm) nanoparticles (microgels) have been synthesized by seed and feed precipitation polymerization, and the influence of chemical differentiation between the core and shell polymers on the phase transition kinetics and thermodynamics has been examined. The results suggest that the core--shell architecture is a powerful one for the design of colloidal "smart gels" with tunable properties. To examine these materials, differential scanning calorimetry (DSC), (1)H NMR, and temperature programmed photon correlation spectroscopy (TP-PCS) have been employed. These measurements show that the addition of small concentrations of a hydrophobic monomer (butyl methacrylate, BMA) into the particle shell produces large decreases in the rate of thermo-induced particle collapse. Conversely, these low levels of hydrophobic modification do not perturb the thermodynamics of the particle phase transition. When these results are examined in light of previous studies of macroscopic hydrogels, they suggest that the formation of a thin, stable skin layer at the particle exterior during the early stages of particle collapse is the rate limiting factor in particle deswelling. Finally, the hydrophobicity (BMA content) of the shell determines the magnitude of the hydrogel collapse rate, while the thickness of the BMA containing region does not impact the observed kinetics. Together, these results suggest that control over the kinetics of microgel deswelling events can be accomplished simply by modification of the particle periphery, and therefore do not require homogeneous modification of the entire polymer structure. PMID- 11480972 TI - Thermodynamic stabilities of linear and crinkled tapes and cyclic rosettes in melamine--cyanurate assemblies: a model description. AB - In this paper we describe model calculations for the self-assembly of N,N disubstituted melamines 1 and N-substituted cyanuric acid or 5,5-disubstituted barbituric acid derivatives 2 into linear or crinkled tapes and cyclic rosettes via cooperative hydrogen bond formation. The model description considers all possible stereoisomeric tape structures consisting of two to eight different components (270 different species in total) and one cyclic hexameric rosette structure. Furthermore, eight steric parameters (R(12)-R(28)) are included that represent the different types of steric interactions within the assemblies. Most importantly, the model calculations clearly show that the tape/rosette ratio is very sensitive to changes in parameters that directly affect the internal energy of the rosette structure. In this respect, three parameters have been characterized, i.e., the basic equilibrium constant K(0) for the bimolecular association of a melamine and cyanurate, the equilibrium constant K(r)/K(0) for the cyclization of a linear hexamer, and the parameter R(12)-a(Z)b, representing attractive or repulsive interactions between adjacent melamine and cyanurate moieties. For example, an increase in K(0) from 100 to 10,000 M(-1) ([A](0) = [B](0) = 10 mM, K(r) = 0.01 M) or in K(r) from 0.001 to 0.1 M ([A](0) = [B](0) = 10 mM, K(0) = 1000 M(-1)) raises the concentration of the rosette from <5 to approximately 90% or from approximately 10 to approximately 85%, respectively. Similarly, a change in R(12)-a(Z)b from 1.0 (no repulsive or attractive interactions) to 1.5 (slight attractive interaction) raises the rosette fraction of the mixture from 25% to 45%. In sharp contrast to this, the model calculations show that parameters that only affect the internal energy of the tapes (R(13)- R(28)) hardly change the tape/rosette ratio. For example, by changing R(13) a(EE)a from 1.0 (no repulsive or attractive interactions) to 0.001 (maximum repulsion), the rosette fraction in the mixture changes by no more than 8%. Including all possible sterics that occur only in tapes (i.e., R(13)--R(28)), the maximum change in rosette fraction is no more than 16%. These predictions can be rationalized by considering that any change in the stability of the tapes only affects the rosette concentration by means of shifting the equilibrium between free 1 and 2 and the rosette. Since there are 270 different tapelike structures in equilibrium, this mixture represents the best buffer solution in the world. These model calculations seem to conflict with the concept of peripheral crowding as put forward by Whitesides et al., which states that bulky substituents on the periphery of the melamine (and cyanurate) components can be used to shift the tape/rosette equilibrium completely toward the rosette structure. Computer simulations (CHARMm 24.0) show that linear tapes with bulky substituents are severely distorted from planarity, while the corresponding rosette remains planar. Therefore, tapelike structures with bulky substituents are expected to have a much higher solubility than the corresponding rosettes, which can explain the observed crystal data. PMID- 11480973 TI - Total synthesis of tricyclic azaspirane derivatives of tyrosine: FR901483 and TAN1251C. AB - A solution to the long-standing problem presented by the oxidative cyclization of a phenolic 3-arylpropionamide to a spirolactam has been developed in this laboratory via oxazoline chemistry. This research was motivated by our interest in some novel tricyclic azaspirane natural products formally derived from tyrosine, such as FR901483 and TAN1251C. In this paper, we disclose full details of the total synthesis of these substances. PMID- 11480974 TI - Self-assembly and anion encapsulation properties of cavitand-based coordination cages. AB - Two novel classes of cavitand-based coordination cages 7a--j and 8a--d have been synthesized via self-assembly procedures. The main factors controlling cage self assembly (CSA) have been identified in (i) a P--M--P angle close to 90 degrees between the chelating ligand and the metal precursor, (ii) Pd and Pt as metal centers, (iii) a weakly coordinated counterion, and (iv) preorganization of the tetradentate cavitand ligand. Calorimetric measurements and dynamic (1)H and (19)F NMR experiments indicated that CSA is entropy driven. The temperature range of the equilibrium cage-oligomers is determined by the level of preorganization of the cavitand component. The crystal structure of cage 7d revealed the presence of a single triflate anion encapsulated. Guest competition experiments revealed that the encapsulation preference of cages 7b,d follows the order BF(4)(-) > CF(3)SO(3)(-) >> PF(6)(-) at 300 K. ES-MS experiments coupled to molecular modeling provided a rationale for the observed encapsulation selectivities. The basic selectivity pattern, which follows the solvation enthalpy of the guests, is altered by size and shape of the cavity, allowing the entrance of an ancillary solvent molecule only in the case of BF(4)(-). PMID- 11480975 TI - De novo design, synthesis, and characterization of antimicrobial beta-peptides. AB - beta-Peptides are a class of polyamides that have been demonstrated to adopt a variety of helical conformations. Recently, a series of amphiphilic L(+2) helical beta-peptides were designed, which were intended to mimic the overall physicochemical properties of a class of membrane-active antimicrobial peptides, including magainin and cecropin. Although these peptides showed potent antimicrobial activity, they also showed significant activity against human erythrocytes. Operating under the assumption that their lack of specificity arose from excessive hydrophobicity, two additional beta-peptides H-(beta(3)-HAla beta(3)-HLys-beta(3)-HVal)(n)-NH(2) (n = 4, 5) were designed and synthesized. Both have high antimicrobial activities, but very low hemolytic potencies. The peptides bind in an L(+2) conformation to phospholipid vesicles, inducing leakage of entrapped small molecules. The peptides have a low affinity for membranes consisting of neutral phosphatidylcholine lipids, but bind avidly to vesicles containing 10 mol % of acidic phosphatidylserine lipids. Differences in vesicle leakage kinetics for the two peptides suggest that chain length could affect their mechanisms of disrupting cell membranes. Thus, insights gained from the study of variants of natural alpha-peptides have provided a useful guide for the design of nonnatural antimicrobial beta-peptides. PMID- 11480976 TI - (1)H NMR investigation of solvent effects in aromatic stacking interactions. AB - One of the marquis challenges in modern Organic Chemistry concerns the design and synthesis of abiotic compounds that emulate the exquisite complex structures and/or functions of biological macromolecules. Oligomers possessing the propensity to adopt well-defined compact conformations, or foldamers, have been attained utilizing hydrogen bonding, torsional restriction, and solvophobic interactions.(1) In this laboratory, aromatic electron donor--acceptor interactions have been exploited in the design of aedamers--foldamers that adopt a novel, pleated secondary structure in aqueous solution. Herein is reported detailed (1)H NMR binding studies of aedamer monomers that were carried out in solvents and solvent mixtures covering a broad polarity range. Curve-fitting analysis of the binding data using a model that incorporated the formation of higher order and self-associated complexes yielded a linear free energy relationship between the free energy of complexation and the empirical solvent polarity parameter, E(T)(30). From these studies, the association of electron rich and electron-deficient aedamer monomers was seen to be driven primarily by hydrophobic interactions in polar solvents. However, the magnitude of these interactions is modulated to a significant extent by the geometry of the donor- acceptor complex, which, in turn, is dictated by the electrostatic complementarity between the electron-deficient and electron-rich aromatic faces of the monomers. PMID- 11480977 TI - Kinetics and mechanism of the beta- to alpha-CuAlCl(4) phase transition: a time resolved (63)Cu MAS NMR and powder X-ray diffraction study. AB - The beta and alpha phases of CuAlCl(4) have been characterized by solid-state (27)Al and (63)Cu magic angle spinning nuclear magnetic resonance. The very short spin--lattice relaxation times of the copper spins, and the sensitivity of the I = 3/2 (63)Cu nucleus to the small differences in the local structure of Cu in the two phases, allowed (63)Cu spectra to be acquired in very short time periods (1 min), in which the beta and alpha phases were clearly resolved. This time resolution was exploited to follow the phase transition from the pseudohexagonal close-packed beta-CuAlCl(4) into the pseudocubic close-packed alpha-CuAlCl(4), which occurs above 100 degrees C. In situ time-resolved (63)Cu MAS NMR and synchrotron X-ray diffraction experiments were used to measure the kinetics of this phase transition as a function of temperature. The transformation was shown to be a first-order phase transition involving no intermediate phases with an activation energy of 138 kJ/mol. The kinetic data obey a first-order Avrami- Erofe'ev rate law. A one-dimensional growth mechanism is proposed that involves a combination of Cu(+) ion self-diffusion and a translational reorganization of the close-packed anion layers imposed by the periodic rotations of [AlCl(4)](-) tetrahedra. This beta to alpha phase transformation can be induced at ambient temperatures by low partial pressures of ethylene. PMID- 11480978 TI - A theoretical study of the aminolysis reaction of lysine 199 of human serum albumin with benzylpenicillin: consequences for immunochemistry of penicillins. AB - Herein, we present results of a computational study on benzylpenicillin attachment to Lys199 of human serum albumin via an aminolysis reaction. The internal geometry of the reactive part of the system was taken from previous work at the B3LYP/6-31+G* level on the water-assisted aminolysis reaction of a penicillin model compound (Diaz, N.; Suarez, D.; Sordo, T. L. J. Am. Chem. Soc. 2000, 122, 6710--6719). The protein environment around Lys199, the 6-acylamino side chain, and the 2-methyl groups of benzylpenicillin were relaxed by carrying out geometry optimizations with a hybrid QM/MM method (PM3/AMBER). Two different mechanistic routes were explored: a one-step water-assisted process and a carboxylate and water-assisted route in which the beta-lactam carboxylate and the ancillary water molecule mediate the proton transfer from the epsilon-amino group of Lys199 to the beta-lactam leaving N atom. The corresponding energy profiles in the protein combine the B3LYP/6-31+G* and PM3 energies of the reactive subsystem (benzylpenicillin + Lys199 side chain + the ancillary water molecule) and semiempirical PM3 energies of the entire system evaluated with a "divide and conquer" linear-scaling method. It is observed that penicillin haptenation to HSA can proceed through the water-assisted concerted mechanism which is calculated to have a high energy barrier of approximately 38 kcal/mol, in agreement with the experimentally observed slow reaction kinetics. PMID- 11480979 TI - Ligand-stabilized ruthenium nanoparticles: synthesis, organization, and dynamics. AB - The decomposition of the ruthenium precursor Ru(COD)(COT) (1, COD = 1,5 cyclooctadiene; COT = 1,3,5-cyclooctatriene) in mild conditions (room temperature, 1--3 bar H(2)) in THF leads, in the presence of a stabilizer (polymer or ligand), to nanoparticles of various sizes and shapes. In THF and in the presence of a polymer matrix (Ru/polymer = 5%), crystalline hcp particles of uniform mean size (1.1 nm) homogeneously dispersed in the polymer matrix and agglomerated hcp particles (1.7 nm) were respectively obtained in poly(vinylpyrrolidone) and cellulose acetate. The same reaction, carried out using various concentrations relative to ruthenium of alkylamines or alkylthiols as stabilizers (L = C(8)H(17)NH(2), C(12)H(25)NH(2), C(16)H(33)NH(2), C(8)H(17)SH, C(12)H(25)SH, or C(16)H(33)SH), leads to agglomerated particles (L = thiol) or particles dispersed in the solution (L = amine), both displaying a mean size near 2--3 nm and an hcp structure. In the case of amine ligands, the particles are generally elongated and display a tendency to form worm- or rodlike structures at high amine concentration. This phenomenon is attributed to a rapid amine ligand exchange at the surface of the particle as observed by (13)C NMR. In contrast, the particles stabilized by C(8)H(17)SH are not fluxional, but a catalytic transformation of thiols into disulfides has been observed which involves oxidative addition of thiols on the ruthenium surface. All colloids were characterized by microanalysis, infrared spectroscopy after CO adsorption, high resolution electron microscopy, and wide-angle X-ray scattering. PMID- 11480980 TI - Argentophilicity and solvent-induced structural diversity in double salts of silver acetylide with silver perfluoroalkyl carboxylates. AB - A series of novel double salts of silver(I) were isolated by dissolving Ag(2)C(2) in a concentrated aqueous solution of R(F)CO(2)Ag (R(F) = CF(3), C(2)F(5)) and AgBF(4). Different ancillary solvento ligands such as H(2)O, CH(3)CN, and C(2)H(5)CN were found to affect the crystallization process that led to the assembly of various silver(I) cages with embedded C(2)(2-) ions. 2Ag(2)C(2) x 12CF(3)CO(2)Ag x 5H(2)O (1) consists of two independent C(2)@Ag(7) cages, each having the shape of a basket with a square base. Ag(2)C(2) x 6CF(3)CO(2)Ag x 3CH(3)CN (2) contains a zigzag chain of edge-sharing triangulated dodecahedra, and 4Ag(2)C(2) x 23CF(3)CO(2)Ag x 7C(2)H(5)CN x 2.5H(2)O (3) features an unusual double-walled silver column constructed from the fusion of four different kinds of irregular polyhedra. Ag(2)C(2) x 10C(2)F(5)CO(2)Ag x 9.5H(2)O (4), Ag(2)C(2) x 9C(2)F(5)CO(2)Ag x 3CH(3)CN x H(2)O (5), and Ag(2)C(2) x 6C(2)F(5)CO(2)Ag x 2C(2)H(5)CN (6) all contain an edge-sharing double cage with each single cage in the shape of a square antiprism, a capped square antiprism, and a triangulated dodecahedron, respectively. PMID- 11480981 TI - pH-dependent modulation of relaxivity and luminescence in macrocyclic gadolinium and europium complexes based on reversible intramolecular sulfonamide ligation. AB - A series of macrocyclic Eu, Gd, and Tb complexes has been prepared in which the intramolecular ligation of a beta-arylsulfonamide nitrogen is rendered pH dependent, giving rise to changes in the hydration state, q, at the lanthanide center. In complexes based on DO3A, variation of the p-substituent in the arylsulfonamide moiety determines the apparent protonation constant log K(MLH) with values of 5.7, 6.4, and 6.7 for the -CF(3), -Me, and -OMe substituents, respectively. Introduction of three beta-carboxyalkyl substituents, alpha to three ring nitrogens, inhibits displacement of the bound water by added protein and also suppresses intermolecular binding by endogenous anions (lactate, HCO(3)( )). Measurements of the pH dependence of the form and intensity of the Eu complexes revealed that intramolecular carboxylate coordination occurred competitively. This was reduced either by enhancing the electron density at the sulfonamide nitrogen or by enlarging the chelate ring from 7--8. Amplification of the relaxivity changes in the pH range 8--5 occurred on protein binding, and over the pH range 7.4--6.8 a 48% change in relaxivity was defined for [Gd.3a] (298 K, 65.6 MHz) in 50% human serum solution. PMID- 11480982 TI - Chemically induced magnetism and magnetoresistance in La(0.8)Sr(1.2)Mn(0.6)Rh(0.4)O(4). AB - It is shown by magnetometry and microSR spectroscopy that short-range magnetic interactions between the Mn cations in the nonmetallic K(2)NiF(4)-like phase La(0.8)Sr(1.2)Mn(0.6)Rh(0.4)O(4) become significant below approximately 200 K. Negative magnetoresistance (rho/rho(0) approximately 0.5 in 14 T at 108 K) is apparent below this temperature. Neutron diffraction has shown that an applied magnetic field of 5 T is sufficient to induce saturated (3.38(7)mu(B) per Mn) long-range ferromagnetic ordering of the atomic moments at 2 K, and that the induced ordering persists up to a temperature of 50 K in 5 T. Spin glass behavior is observed below 20 K in the absence of an applied field. The induced magnetic ordering is attributed to the subtle changes in band structure brought about by the external field, and to the controlling influence of Rh(3+) over the relative strength of competing magnetic exchange interactions. PMID- 11480983 TI - Theoretical (DFT) insights into the mechanism of copper-catalyzed cyclopropanation reactions. Implications for enantioselective catalysis. AB - The mechanism of the copper(I)-catalyzed cyclopropanation reaction has been extensively investigated for a medium-size reaction model by means of B3LYP/6 31G(d) calculations. The starting ethylene complex of the N,N' dimethylmalonaldiimine--copper (I) catalyst undergoes a ligand exchange with methyl diazoacetate to yield a reaction intermediate, which subsequently undergoes nitrogen extrusion to generate a copper--carbene complex. The cyclopropanation step takes place through a direct carbene insertion of the metal -carbene species to yield a catalyst--product complex, which can finally regenerate the starting complex. The stereochemical predictions of a more realistic model (by considering a chiral bis(oxazoline)--copper (I) catalyst) have been rationalized in terms of steric repulsions, showing good agreement with experimental data. PMID- 11480984 TI - Inhibition of DNA transcription using cationic mixed monolayer protected gold clusters. AB - Efficient recognition of DNA is a prerequisite for the development of biological effectors, including transcription and translation regulators, transfection vectors, and DNA sensors. To provide an effective scaffold for multivalent interactions with DNA, we have fabricated mixed monolayer protected gold clusters (MMPCs) functionalized with tetraalkylammonium ligands that can interact with the DNA backbone via charge complementarity. Binding studies indicate that the MMPCs and DNA form a charge-neutralized, nonaggregated assembly. The interactions controlling these assemblies are highly efficient, completely inhibiting transcription by T7 RNA polymerase in vitro. PMID- 11480985 TI - Unraveling the origin of regioselectivity in rhodium diphosphine catalyzed hydroformylation. A DFT QM/MM study. AB - The origin of regioselectivity in rhodium diphosphine catalyzed hydroformilation was investigated by means of hybrid QM/MM calculations using the IMOMM method. The roles of the diphosphine bite angle and of the nonbonding interactions were analyzed in detail by considering rhodium systems containing xantphos-type ligands, for which a correlation between the natural bite angle and regioselectivity has been recently reported. From the pentacoordinated equatorial -equatorial HRh(CO)(alkene)(diphosphine) key intermediate, eight possible reaction paths were defined and characterized through their respective transition states (TS). We succeeded in reproducing the experimentally observed trends for the studied diphosphines. By performing additional calculations on model systems, in which the steric effects induced by the phenyl substituents of xantphos ligands were canceled, we were able to separate, identify, and evaluate the different contributions to regioselectivity. These additional calculations showed that regioselectivity is governed by the nonbonding interactions between the diphenylphosphino substituents and the substrate, whereas the effects directly associated to the bite angle, what we call orbital effects, seem to have a smaller influence. PMID- 11480986 TI - Semiclassical simulations of azomethane photochemistry in the gas phase and in solution. AB - We present semiclassical simulations of the dynamical events which follow the n - > pi excitation of an azomethane molecule: decay to the ground state, isomerization, and dissociation. Ab initio potential energy surfaces and couplings, slightly modified according to available experimental data, are employed in conjunction with a trajectory-surface-hopping method. For an isolated molecule, we show that dissociation takes place in the ground state, because the radiationless decay is very fast. The dissociation is mainly sequential: the methyldiazenyl radical first produced fragmentates very rapidly to N(2) + CH(3)*. The results of previous experiments by Lee's and Zewail's groups, from which partially conflicting conclusions had been drawn, are reproduced and interpreted. Molecular dynamics, coupled to the surface-hopping algorithm, allows us to simulate the photochemistry in acqueous solution. Here the fragmentation is suppressed because of vibrational energy loss to the solvent: only the isomerization takes place. PMID- 11480987 TI - A two-dimensional (magnetic field and concentration) electron paramagnetic resonance method for analysis of multispecies complex equilibrium systems. information content of EPR spectra. AB - A two-dimensional simulation method has been developed for the interpretation of electron paramagnetic resonance (EPR) spectra consisting of a multitude of strongly overlapping signal components. The set of EPR spectra for complex equilibrium systems is analyzed simultaneously as a function of metal and ligand concentrations and pH. The formation constants of the various species are adjusted together with the magnetic parameters of the component EPR spectra. At most 10 EPR-active and 5 EPR-silent species can be involved to simulate a maximum of 36 experimental spectra, while the number of adjusted parameters is at most 100. Statistical parameters are suggested to give the confidence intervals for parameter estimation and to distinguish alternative speciation models. The efficiency of the program is demonstrated for the copper(II)--L-asparagine system, in which 10 species, including 3 pairs of isomers, are characterized with magnetic parameters and formation constants. On the basis of the magnetic parameters, a structural assignment is made for the detected species. The two dimensional approach can also supply the formation constant of the EPR-silent species, as demonstrated for the copper(II)--glycyl-L-serine system. PMID- 11480988 TI - A periodic DFT study of intramolecular isomerization reactions of toluene and xylenes catalyzed by acidic mordenite. AB - A periodic density functional theory (DFT) study of the isomerization reactions of toluene and xylene catalyzed by acidic mordenite is reported. Monomolecular isomerization reactions have been considered and analyzed. The different reaction pathways have been discussed in detail. The use of periodic structure calculations allows consideration and analysis of zeolite electrostatic contributions and steric constraints that occur within zeolite micropores. Major differences in the details of protonation reaction pathways are found when periodic structures are used rather than small cluster models of the Bronsted acidic site. Complex relationships are found between zeolite topology and reaction pathways. PMID- 11480989 TI - Intramolecular energy hopping and energy trapping in polyphenylene dendrimers with multiple peryleneimide donor chromophores and a terryleneimide acceptor trap chromophore. AB - Intramolecular Forster-type excitation energy transfer (FRET) processes in a series of first-generation polyphenylene dendrimers substituted with spatially well-separated peryleneimide chromophores and a terryleneimide energy-trapping chromophore at the rim were investigated by steady-state and time-resolved fluorescence spectroscopy. Energy-hopping processes among the peryleneimide chromophores are revealed by anisotropy decay times of 50--80 ps consistent with a FRET rate constant of k(hopp) = 4.6 ns(-1). If a terryleneimide chromophore is present at the rim of the dendrimer together with three peryleneimide chromophores, more than 95% of the energy harvested by the peryleneimide chromophores is transferred and trapped in the terryleneimide. The two decay times (tau(1) = 52 ps and tau(2) = 175 ps) found for the peryleneimide emission band are recovered as rise times at the terryleneimide emission band proving that the energy trapping of peryleneimide excitation energy by the terryleneimide acceptor occurs via two different, efficient pathways. Molecular- modeling-based structures tentatively indicate that the rotation of the terryleneimide acceptor group can lead to a much smaller distance to a single donor chromophore, which could explain the occurrence of two energy-trapping rate constants. All energy transfer processes are quantitatively describable with Forster energy transfer theory, and the influence of the dipole orientation factor in the Forster equation is discussed. PMID- 11480990 TI - Self-assembly of 10-microm-sized objects into ordered three-dimensional arrays. AB - This paper describes the self-assembly of small objects--polyhedral metal plates with largest dimensions of 10 to 30 microm--into highly ordered, three dimensional arrays. The plates were fabricated using photolithography and electrodeposition techniques, and the faces of the plates were functionalized to be hydrophobic or hydrophilic using self-assembled monolayers (SAMs). Self assembly occurs in water through capillary interactions between thin films of a hydrophobic liquid (a liquid prepolymer adhesive) coated onto the hydrophobic faces of the plates; coalescence of the adhesive films reduces the interfacial free energy of the system and drives self-assembly. By altering the size and surface-patterning of the plates, the external morphologies of the aggregates were varied. Curing the adhesive furnished mechanically stable aggregates that were characterized by scanning electron microscopy (SEM). For assemblies formed by plates partially composed of a sacrificial material, a subsequent etching step furnished fully open, three-dimensional microstructures. This work validates the use of capillary interactions for three-dimensional mesoscale self-assembly in the 10-microm-size regime and opens new avenues for the fabrication of complex, three-dimensional microscructures. PMID- 11480991 TI - Solvent-dependent transition states for decarboxylations. AB - The rate constants and kinetic isotope effects for decarboxylation of 4 pyridylacetic acid depend strongly on whether the solvent is water or dioxane, and the present paper interprets this finding. We calculate the solvent dependence of the free energy barrier and of the (13)C and (18)O kinetic isotope effects using a quantum mechanical solvation model based on class IV charges and semiempirical atomic surface tensions. The calculations provide a consistent interpretation of the experimental results, which provides a striking confirmation of the soundness of the solvation modeling. Even more significantly, the agreement of theory and experiment gives us confidence in the physical picture of the reaction provided by the model. This indicates that the location of the transition state, as measured by the length of the breaking C--C bond, is 0.24 A later than the gas phase in dioxane and 0.37 A later than the gas phase in water. Charge development at the transition state also depends strongly on the solvent; in particular the CO(2) moiety is 0.07 electronic charge units more negative at the transition state in dioxane than in water. PMID- 11480992 TI - Quantum chemical study of ester aminolysis catalyzed by a single adenine: a reference reaction for the ribosomal peptide synthesis. AB - Herein we report the results of a HF/6-31+G** and B3LYP/6-31+G** computational investigation of the title reaction for the peptide bond synthesis catalyzed by a single adenine. This system constitutes a reference reaction to study the basic chemical events that have been proposed to occur at the peptidyl transferase active site of ribosomes on the basis of structural determinations (Science 2000, 239, 920--931). Thus, the analysis of the geometry, charge distribution, and energetics of the critical structures involved in this title reaction yields insight into the catalytic mode of action of RNA molecules. Our computational results give further support to the hypotheses that the activated nucleotide A2451 in the ribosome acts as a base catalyst and that this role is similar to that of the His residue in the catalytic triad of serine proteases. PMID- 11480993 TI - Ion conductors derived from cholic acid and spermine: importance of facial hydrophilicity on NA(+) transport and membrane selectivity. AB - A series of ion conductors have been synthesized in which the degree of facial hydrophilicity has been systematically varied. Specifically, conjugates have been prepared from cholic acid and spermine in which the hydrophilic face of each sterol bears methoxy (1), hydroxy (2), carbamate (3), or sulfate groups (4). The ability of these conjugates to promote the transport of Na(+) across phosphatidylcholine membranes of varying thickness has been investigated by (23)Na NMR spectroscopy. Examination of observed activities in three different phosphatidylcholine membranes has provided evidence for membrane-spanning dimers as the transport-active species. In the thinnest membranes investigated, made from 1,2-dimyristoleoyl-sn-glycero-3-phosphocholine (C14), Na(+)-transport activity was found to increase, substantially, with increasing facial hydrophilicity. In thicker membranes, made from 1,2-dioleoyl-sn-glycero-3 phosphocholine (C18), observed activities were found to decrease with increasing facial hydrophilicity; with a membrane of intermediate thickness, prepared from 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine (C16), ion-conducting activity increased and then decreased, with continuous increases in facial hydrophilicity. The possible origins for these variations in activity are briefly discussed. PMID- 11480994 TI - Potential-dependent vibrational spectroscopy of solvent molecules at the Pt(111) electrode in a water/acetonitrile mixture studied by sum frequency generation. AB - Sum frequency generation (SFG) vibrational spectra of D(2)O and/or acetonitrile (CH(3)CN) on a Pt(111) single-crystal electrode were obtained as a function of applied potential in a 5 mol % water/acetonitrile mixed solvent with different 0.1 molar MSO(3)CF(3) salts (M = H(+), Li(+), Na(+), K(+), and Cs(+)). The results provide a very specific model for the composition of the inner Helmholtz layer as a function of potential and surface charge. Acetonitrile dominates the inner layer with the CN group directed toward the metal at potentials where the metal has a positive charge. As the surface becomes negatively charged, the acetonitrile orientation flips 180 degrees, with the CH(3) group pointing toward the surface. At even more negative surface charge, D(2)O displaces acetonitrile from the inner layer and is the predominant molecule on the surface. Here water is present as an oriented molecule with the oxygen end pointing toward the metal. The potential (and surface charge) where water is the dominant molecule in the inner Helmholtz layer is determined by the solvation energy of the cation. PMID- 11480995 TI - A contribution to the design of molecular switches: novel acid-mediated ring closing--photochemical ring-opening of 2,3-bis(heteroaryl)quinones (heteroaryl = thienyl, furanyl, pyrrolyl). PMID- 11480996 TI - Guanidinium ylides as a new and recyclable source for aziridines and their roles as chiral auxiliaries. PMID- 11480997 TI - Copper-catalyzed nitrogen transfer mediated by iodosylbenzene PhI=O. PMID- 11480998 TI - The energetic contribution of backbone--backbone hydrogen bonds to the thermodynamic stability of a hyperstable P22 Arc repressor mutant. PMID- 11480999 TI - Double deprotonation of a cyclopentadienyl alkene to form a polydentate trianionic cyclopentadienyl allyl ligand system. PMID- 11481000 TI - Main group "constrained geometry" complexes. PMID- 11481001 TI - Hydroperoxo--copper(II) complex stabilized by N(S)s-type ligand having a phenyl thioether. PMID- 11481002 TI - Acid catalysis in HZSM-5: the role of entropy. PMID- 11481003 TI - A novel one-pot reaction: zwitterionic rhodium complex-catalyzed hydroaminovinylation of vinyl sulfones and a vinylphosphonate. PMID- 11481004 TI - Diversity in short beta-peptide 12-helices: high-resolution structural analysis in aqueous solution of a hexamer containing sulfonylated pyrrolidine residues. PMID- 11481005 TI - Polystyrene bead-assisted self-assembly of microstructured silica hollow spheres in highly alkaline media. PMID- 11481006 TI - The palladium-catalyzed oxidative kinetic resolution of secondary alcohols with molecular oxygen. PMID- 11481007 TI - A general and efficient copper catalyst for the amidation of aryl halides and the N-arylation of nitrogen heterocycles. PMID- 11481009 TI - A stable halosilylene at room temperature in THF solution. PMID- 11481008 TI - A conducting polymer nanojunction switch. PMID- 11481010 TI - Novel [2,3]-sigmatropic rearrangement for carbon--nitrogen bond formation. PMID- 11481011 TI - The first structurally characterized hypervalent silicon hydride: unexpected molecular geometry and Si--H...K interactions. PMID- 11481012 TI - Nanorainbows: graded semiconductor films from quantum dots. PMID- 11481013 TI - Discrete supramolecular architecture vs crystal engineering: the rational design of a platinum-based bimetallic assembly with a chairlike structure and its infinite, copper analogue. PMID- 11481014 TI - Understanding the role of helical chains in the formation of noncentrosymmetric solids. PMID- 11481016 TI - Quality of life (QOL) versus curability for lung cancer surgery. AB - Standard operations for lung cancer patients are generally accepted as performing lobectomy or pneumonectomy on the tumor bearing lung and ipsilateral hilar and mediastinal lymphadenectomy including subcarinal lymph nodes. Recently, minimally invasive surgery or limited resection (for example, those via VATS) has ruled our time in the field of surgery considering especially from the point of QOL. There are so many factors that cause any decline to lung cancer patients' postoperative QOL, such as operative death, postoperative cancer death, postoperative complications, long-lasting discomfort symptoms and so forth. However, a surgery, even though it is big or extensive, does not always inevitably reduce QOL for patients with lung cancer. If patients received curable resection and have got cured, it seems that they would almost all be satisfied with their postoperative QOL. Namely, at present, we do not give priority to QOL but we should give priority to curability for lung cancer surgery, if the patients have no special risk factors, which eventually would bring them almost satisfactory postoperative QOL. PMID- 11481017 TI - A study on the role of platelet function in patients with chronic pulmonary thromboembolism. AB - There has been no study on the platelet function in the patient with chronic pulmonary thromboembolism (CPTE). We speculate that the platelet function may be elevated in the patients. PURPOSE: 1. The platelet functions were compared among CPTE before surgery, deep vein thrombosis (DVT) and normal adult people. 2. The severity of CPTE in clinical grading to the platelet functions were compared. 3. The platelet function were compared before and after pulmonary thromboendarterectomy. METHODS: Pre-opetative CPTE group (n=16), post-operative CPTE group (n=11), DVT group (n=9) and control group (normal adult people: n=33) were investigated on the platelet functions defined as platelet adhesion (AD) and platelet aggregation (AG) test in this study. RESULTS: 1. No activation of platelet functions was observed in pre-operative CPTE patients. 2. There was no apparent relationship between the severity of disease and platelet functions. 3. Significant elevation of AG was obtained in the patients who received pulmonary thromboendarterectomy. CONCLUSION: In consideration to the finding in postoperative study, the administration of anti-platelet drug will help to prevent re-thrombosis of the pulmonary arteries after surgery. PMID- 11481018 TI - Diagnosis of deep vein thrombosis using platelet scintigraphy. AB - PURPOSE: We reviewed the usefulness of platelet scintigraphy, in which autologous platelets labeled with indium-111-oxine reveal thrombotic activity, for evaluating deep vein thrombosis (DVT). MATERIALS: During the past 2 years, 39 cases with DVT were enrolled in this study. DVT was definitely diagnosed by color duplex scanning, platelet scintigraphy or both in all cases. For semiquantitative analysis, we estimated the ratio of accumulation in the abnormal region to that in the normal vein on the other side, and defined an abnormal accumulation ratio as over 1.2. RESULTS: Abnormal accumulation ratio showing active DVT was recognized in 30 cases (77%), and showed a good correlation with clinical symptoms. In addition, in 19 cases with crural DVT, platelet scintigraphy showed abnormal accumulation ratio in 16 cases (84%), while duplex scanning detected thrombi in 13 cases (68%). In cases with abnormal accumulation ratio, thrombolytic and anticoagulant therapy were very effective for improving clinical symptoms as well inducing regression of the accumulation ratio. CONCLUSIONS: Platelet scintigraphy was very useful for the diagnosis and treatment of DVT and for evaluation of the effect of anticoagulant therapy. Limitations in the definite diagnosis of deep vein thrombosis (DVT) have been apparent for more than three decades. During the last decade, duplex scanning has reached a high level of accuracy and has been considered the gold standard in the diagnosis of DVT, instead of venography. However, duplex scanning and venography demonstrates only the anatomic alterations associated with venous lesions. In contrast, in platelet scintigraphy, the labeled platelets are incorporated directly into the thrombus and can reveal thrombus activity. We noticed that autologous platelets labeled with indium-111-oxine accumulated on fresh lesions of DVT. This observation suggested two applications of this technique: 1) evaluation of the role of platelets in the pathophysiologic characteristics of DVT; and 2) monitoring the effects of anticoagulant therapy. PMID- 11481019 TI - Functional, metabolic, and histological changes of vascular tissues after warm ischemia. AB - We examined functional, metabolic, and histological changes in the aortic tissue of rats after the period of warm ischemia ranging from 0 to 24 hours to determine the window of time in which grafts can be optimally viable for harvest. Sixty aortas from Brown Norway rats obtained after warm ischemia were used and changes in contraction, endothelial-dependent or -independent vasodilatation, cell viability, and histology were examined. Maximal contraction induced by norepinephrine and potassium chloride decreased time-dependently after exposure to warm ischemia. The warm ischemic period when 50% of the maximal contractile response of freshly isolated arteries was preserved, ranged from 6 to 8 hours. Maximal endothelium-dependent relaxation induced by acetylcholine decreased along with the time of warm ischemia. Endothelium-independent relaxation induced by sodium nitroprusside and forskolin was unaltered for up to 9 hours. Cell viability gradually decreased, and a significant negative correlation was found between warm ischemic period (T: hours) and cell viability (V: %) (V=101.9-2.35T; r(2)=0.96; p<0.0001). Cell viability was greater than 70% within 12 hours postmortem. Histologically, after 9-hour-warm ischemia irreversible changes were detected. Results suggest that the period of warm ischemia for up to 6 hours would be acceptable for preservation of tissue viability. PMID- 11481020 TI - Favorable results in patients with small size CarboMedics heart valves in the aortic position. AB - Hemodynamic performance of the CarboMedics heart valve in the aortic position and its clinical impacts were investigated in 126 consecutive patients. The actuarial survival rates of patients who had undergone isolated aortic valve replacement and concomitant aortic and mitral valve replacement were 82.6+/-5.7% and 71.0+/ 9.2% at 8 years, respectively. Morbid events were rare, and almost all late survivors were free from evident cardiac symptoms regardless of the valve size. Echocardiography revealed suboptimal transvalvular pressure gradients and effective orifice areas of 19 mm and 21 mm valves. However, relief of the left ventricular overload and improvement of the clinical symptoms as well as cardiac function were comparable to those of patients with larger valves. Valve function measured by echocardiography did not show significant correlation to late outcome. Good results can be expected even in the presence of echocardiographic data such as peak pressure gradient over 40 mmHg, effective orifice area less than 1.0 cm(2), and effective orifice area index less than 0.7 cm(2)/m(2). PMID- 11481021 TI - Free right internal thoracic artery as a second arterial conduit: modification of proximal anastomosis for improvement of graft patency. AB - The applicability of the right internal thoracic artery (RITA) for coronary artery bypass grafting is higher when used as a free graft than as a pedicled graft. However, the technical difficulty of directly connecting the proximal end of the free RITA to the much larger aorta leads to poor patency. To overcome this technical limitation, we have used a modification that places the proximal end of this artery onto the hood of an accompanying vein graft at the aortic anastomosis instead of directly onto the aorta. We performed isolated coronary artery bypass grafting on 43 patients using the free RITA as a second arterial graft following pedicled left internal thoracic artery grafting. The mean patient age was 60 years and 38 patients were male. There was no mortality and no incidence of morbidity related to free RITA use. Postoperative coronary angiography performed in all patients revealed that all proximal anastomoses were widely patent, making the patency rate of the free RITA 100%. With these encouraging results, the free RITA graft with the described modification is thought to be a more promising second arterial graft with greater versatility than the pedicled graft. The long term evaluation of a large patient population will determine the significance of this modification. PMID- 11481022 TI - Bronchial stump reinforcement in right pneumonectomy with fascia lata and gelatin resorcin formalin (GRF) glue: case report. AB - We reinforced the bronchial stump with fascia lata and Gelatin Resorcin Formalin (GRF) glue in a right pneumonectomy. This method was found to be simple and useful. We describe our case and the method herein. A 62-year-old woman had a malignant polypoid lesion which completely occluded the introitus of the right main bronchus and deviated to the introitus of the left main bronchus. Right pneumonectomy was done but materials (pleura, pericardium, intercostal muscle, etc.) obtained from the thoracic cavity were insufficient for bronchial stump reinforcement due to severe adhesion caused by prior tuberculosis. Therefore, we reinforced the bronchial stump using the fascia lata and GRF glue. Fascia lata is a superior material for reinforcement in terms of strength and ease of molding, as well as harvesting. GRF glue is a superior adhesive with rapid and strong fixation. We consider this method of reinforcing the bronchial stump with fascia lata and GRF glue to be feasible, in particular, for pneumonectomy or lobectomy without adequate material in the thoracic cavity because of severe adhesion or lesions. PMID- 11481023 TI - Lung cancer with p53 expression and a solitary metastasis to the stomach: a case report. AB - Lung cancer with a solitary metastasis to the stomach occurred in a 65-year-old man, surgically treated for gastric metastasis was followed by pulmonary resection. The gastric metastasis accompanied by upper gastrointestinal hemorrhage. After total gastrectomy to control this hemorrhage, a left lower lobectomy with a partial resection of the lingular segment and combined resection of the chest wall were done. Histopathological features of both the primary tumor in the left lower lobe and the gastric tumor were poorly differentiated adenocarcinoma, and showed the same immunoreactivities of p53 protein, carcinoembryonic antigen and keratin. These results indicate that the gastric tumor was a metastasis originated from the lung cancer. PMID- 11481024 TI - Right atrial myxoma associated with atrial septal defect: a case report and review of the literature. AB - We describe a case of right atrial myxoma with mild cyanosis due to a right-to left shunt at the atrial level. The patient was a 31-year-old woman with a 10 month history of easy fatigability and shortness of breath. Echocardiography showed a right atrial tumor producing a partial dynamic tricuspid obstruction. Digital subtraction angiography via the superior vena cava disclosed a mass lesion which occupied the right atrium with early visualization of the ascending aorta. Successful excision of the tumor and repair of the atrial septal defect totally relieved her presenting symptoms. In a rare association of a right atrial myxoma with atrial septal defect, preoperative evaluation and operative management are discussed. PMID- 11481025 TI - Aortic valve replacement combined with endoventricular circulatory patch plasty (Dor operation) in a patient with aortic valve stenosis and severe ischemic cardiomyopathy. AB - A 58-year-old woman with ischemic cardiomyopathy and aortic valve stenosis, underwent aortic valve replacement and simultaneous endoventricular circulatory patch plasty (Dor operation). She underwent coronary artery bypass grafting for severe triple vessel disease 10 years ago. Recently she started to show severe congestive heart failure. Aortic valve stenosis with pressure gradient of 85-mmHg was also found. Coronary bypasses were all patent, but the left ventricle (LV) was severely dilated (LVDd/Ds=71/61 mm) and the ischemic cardiomyopathy was considered as the cause. She successfully underwent aortic valve replacement and endoventricular circulatory patch plasty. The initial postoperative course was complicated with intractable ventricular arrhythmia, but subsequent course was smooth and the patient was discharged with improved symptoms (NYHA Class II). Postoperative catheterization showed decreased left ventricular volume and improved contractility. This case implies the role of LV remodeling procedure in the ischemic cardiomyopathy combined with aortic valve lesion PMID- 11481026 TI - Mitral valve plasty for mitral regurgitation after blunt chest trauma. AB - A 21 year-old woman was admitted to our hospital because of chest and back pain after blunt chest trauma. On admission, consciousness was clear and a physical examination showed labored breathing. Her vital signs were stable, but her breathing gradually worsened, and artificial respiration was started. The chest roentgenogram and a subsequent chest computed tomographic scans revealed contusions, hemothorax of the left lung and multiple rib fractures. A transthoracic echocardiography (TTE) revealed normal left ventricular wall motion and mild mitral regurgitation (MR). TTE was carried out repeatedly, and revealed gradually progressive MR and prolapse of the posterior medial leaflet, although there was no congestive heart failure. After her general condition had recovered, surgery was performed. Intraoperative transesophageal echocardiography (TEE) revealed torn chordae at the posterior medial leaflet. The leaflet where the chorda was torn was cut and plicated, and posterior mitral annuloplasty was performed using a prosthetic ring. One month later following discharge, the MR had disappeared on TTE. PMID- 11481027 TI - Transatrial repair of ventricular septal rupture under preoperative localization by transesophageal echocardiography. AB - We report about a 71-year-old woman with postinfarction ventricular septal rupture who was successfully treated by the transatrial closure under preoperative localization by transesophageal echocardiography. In an attempt at transatrial repair of the ventricular septal rupture, the most important thing is preoperative localization of the defect in the septum, which is located high and posterior, where it is smooth with relatively few trabeculations and can be readily exposed by retraction of the tricuspid valve. PMID- 11481028 TI - Sextuple coronary artery bypass grafting using only in situ arterial conduits. AB - Sequential bypass in coronary artery bypass grafting with in situ arterial conduits, the bilateral internal thoracic arteries and the right gastroepiploic artery, is one of the most important procedures using a limited number of in situ arterial conduits to revascularize a wide area, although it demands rather difficult techniques. We report a case of a 50-year-old man who underwent sextuple bypass using only in situ arterial grafts with three sequential anastomoses. PMID- 11481029 TI - Simple and safe cannulation technique for antegrade selective cerebral perfusion. AB - Antegrade Selective Cerebral Perfusion (SCP) has proved to be a safe and reliable method of brain protection during operations of aortic arch aneurysms. Arch vessel cannulation is a crucial step in the institution of SCP that determines its success to a significant extent. A simple and safe cannulation technique along with a newly developed flexible perfusion cannula is described. PMID- 11481030 TI - TNF-alpha induced endothelial MAdCAM-1 expression is regulated by exogenous, not endogenous nitric oxide. AB - BACKGROUND: MAdCAM-1 is an adhesion molecule expressed in Peyer's patches and lymphoid tissues which is mobilized by cytokines like TNF-alpha and is a major determinant of lymphocyte trafficking to the gut in human inflammatory bowel disease (IBD). It has been suggested that both reactive oxygen and nitrogen metabolites participate in regulating adhesion molecule expression in response to TNF-alpha. METHODS: To examine how exogenous and endogenous sources of NO modulate MAdCAM-1 induction by TNF-alpha, we pre-treated mouse lymphatic endothelial cells with either long or short acting NO donors prior to TNF-alpha stimulation, and measured MAdCAM-1 induction at 24 h. RESULTS AND DISCUSSION: DETA-NO, a long-acting NO donor, and SperNO, a rapid releasing NO donor both inhibited TNF-alpha-stimulated MAdCAM-1 expression in a concentration dependent manner. Both NO donors also reduced a4b7-dependent lymphocyte endothelial adhesion. Inhibition of endogenous NO production by either L-NAME, a non selective NOS inhibitor, or by 1400 w, a selective iNOS inhibitor failed to induce, or potentiate TNF-alpha regulated MAdCAM-1 expression. CONCLUSIONS: Exogenous NO donors may be beneficial in the treatment of IBD, while endogenous nitric oxide synthases may be less effective in controlling adhesion molecule expression in response to cytokines. PMID- 11481031 TI - Local and distant recurrences in rectal cancer patients are predicted by the nonspecific immune response; specific immune response has only a systemic effect- a histopathological and immunohistochemical study. AB - BACKGROUND: Invasion and metastasis is a complex process governed by the interaction of genetically altered tumor cells and the immunological and inflammatory host response. Specific T-cells directed against tumor cells and the nonspecific inflammatory reaction due to tissue damage, cooperate against invasive tumor cells in order to prevent recurrences. Data concerning involvement of individual cell types are readily available but little is known about the coordinate interactions between both forms of immune response. PATIENTS AND METHODS: The presence of inflammatory infiltrate and eosinophils was determined in 1530 patients with rectal adenocarcinoma from a multicenter trial. We selected 160 patients to analyze this inflammatory infiltrate in more detail using immunohistochemistry. The association with the development of local and distant relapses was determined using univariate and multivariate log rank testing. RESULTS: Patients with an extensive inflammatory infiltrate around the tumor had lower recurrence rates (3.4% versus 6.9%, p = 0.03), showing the importance of host response against tumor cells. In particular, peritumoral mast cells prevent local and distant recurrence (44% versus 15%, p = 0.007 and 86% versus 21%, p < 0.0001, respectively), with improved survival as a consequence. The presence of intratumoral T-cells had independent prognostic value for the occurrence of distant metastases (32% versus 76%, p < 0.0001). CONCLUSIONS: We showed that next to properties of tumor cells, the amount and type of inflammation is also relevant in the control of rectal cancer. Knowledge of the factors involved may lead to new approaches in the management of rectal cancer. PMID- 11481032 TI - Microglia: activation and their significance in the central nervous system. AB - Microglia are resident monocyte-lineaged cells in the brain. Their characteristic feature is that they react to injury and diseases of the brain and become morphologically and functionally activated. Although some trigger molecules which activate microglia are predicted to be released from injured or affected cells, such molecules have not yet been identified. The main role of activated microglia is believed to be in brain defense, as scavengers of dead cells, and as immune or immunoeffector cells. Recent biochemical and neurobiological studies have further indicated that they significantly affect the pathological state and/or regulate the regenerative state and remodeling of the brain by producing a variety of biologically active molecules including cytotoxic and neurotrophic molecules. PMID- 11481033 TI - Structure and function of Syk protein-tyrosine kinase. AB - Non-receptor type of protein-tyrosine kinase Syk contains 2 Src homology 2 (SH2) domains in tandem and multiple autophosphorylation sites. Syk is activated upon binding of tandem SH2 domains to immunoreceptor tyrosine-based activating motif (ITAM) and plays an essential role in lymphocyte development and activation of immune cells. Syk is critical for tyrosine phosphorylation of multiple proteins which regulate important pathways leading from the receptor, such as Ca(2+) mobilization and mitogen-activated protein kinase (MAPK) cascades. Recent findings reveal that expression of Syk appears to be involved in a wide variety of cellular functions and pathogenesis of malignant cancer. These observations have demonstrated that Syk is a key molecule that controls multiple physiological functions in cells. PMID- 11481034 TI - Identification of a new imprinted gene, Rian, on mouse chromosome 12 by fluorescent differential display screening. AB - Systematic screening of differentially expressed genes among androgenetic, parthenogenetic, and normal embryos by means of fluorescent differential display revealed five imprinted genes. One of them, named Rian, was expressed exclusively from the maternal allele and was closely linked to an imprinted gene, Meg3(Gtl2), mapped to the distal end of chromosome 12. The Rian transcript did not have any apparent open reading frame, and its transcript was exclusively localized to the nucleus. PMID- 11481035 TI - X-ray absorption spectroscopic analysis of the high-spin ferriheme site in substrate-bound neuronal nitric-oxide synthase. AB - Nitric oxide synthase (NOS) catalyzes the conversion of L-arginine to citrulline and nitric oxide through two stepwise oxygenation reactions involving N(omega) hydroxy-L-arginine, an enzyme-bound intermediate. The N(omega)-hydroxy-L-arginine and arginine-bound NOS ferriheme centers show distinct, high-spin electron paramagnetic resonance signals. Iron X-ray absorption spectroscopy (XAS) has been used to examine the structure of the ferriheme site in the N(omega)-hydroxy-L arginine-bound full-length neuronal NOS in the presence of (6R)-5,6,7,8 tetrahydro-L-biopterin. Iron XAS shows that the high-spin ferriheme sites in the N(omega)-hydroxy-L-arginine- and arginine-bound forms are strikingly similar, both being coordinated by the heme and an axial thiolate ligand, with an Fe-S distance of ca. 2.29 A. Cu(2+) inhibition slightly affects the spin-state equilibrium, but causes no XAS-detectable changes in the immediate ferriheme coordination environment of neuronal NOS. The structure and ligand geometry of the high-spin ferriheme in arginine-bound neuronal NOS are essentially identical to those of the N(omega)-hydroxy-L-arginine-bound form and only slightly affected by the divalent cation inhibitor of constitutive NOS. PMID- 11481036 TI - The carboxyl terminal sequence of nucleolar protein B23.1 is important in its DNA polymerase alpha-stimulatory activity. AB - The protein B23 is a major nucleolar phosphoprotein comprising two isoforms, B23.1 and B23.2, which differ only in their carboxyl-terminal short sequences, the N-terminal 255 residues being identical in both forms. Both B23.1 and B23.2 stimulated immunoaffinity-purified calf thymus DNA polymerase alpha in a dose dependent manner. The stimulatory effect of protein B23.1, the longer isoform, was found to be 2-fold greater than that of B23.2. Purified DNA polymerase alpha bound tightly to a protein B23.1-immobilized column, while it bound weakly to a protein B23.2-immobilized column. Surface plasmon resonance studies by BIAcore further showed that protein B23.1 bound to the DNA polymerase alpha-(dA).(dT) complex more tightly than did protein B23.2. The protein B23 isoforms appear to interact directly with the DNA polymerase alpha protein and not through the bound nucleic acid. These observations indicated that protein B23 physically bound to the DNA polymerase alpha and stimulated the enzyme activity. Product analyses showed that protein B23 greatly enhanced the reaction both in amount and length of product DNA, whereas it did not significantly alter the processivity of polymerization. In contrast, protein B23 effectively protected DNA polymerase alpha from heat inactivation. These results suggest that protein B23 stabilizes DNA polymerase alpha that is detached from product DNA, allowing the enzyme to be recruited for further elongation. Moreover, experiments using various C-terminal deletion mutants of protein B23 indicated that 12 amino acids at the C-terminal end of B23.1, which are absent in B23.2, may be essential for the full stimulation of the DNA polymerase alpha. PMID- 11481037 TI - Identification and characterization of two penta-EF-hand Ca(2+)-binding proteins in Dictyostelium discoideum. AB - Penta-EF-hand (PEF) proteins such as ALG-2 (apoptosis-linked gene 2 product) and the calpain small subunit are a newly classified family of Ca(2+)-binding proteins that possess five EF-hand-like motifs. We identified two mutually homologous PEF proteins, designated DdPEF-1 and DdPEF-2 (64% amino acid residue identities), in the cellular slime mold Dictyostelium discoideum. Both PEF proteins showed a higher similarity to mammalian ALG-2 and peflin (Group I PEF proteins) than to calpain and sorcin subfamily (Group II PEF proteins) in the first EF-hand (EF-1) regions. Northern blot analyses revealed that DdPEF-1 and DdPEF-2 were constitutively expressed throughout development of Dictyostelium, but their levels of expression were developmentally regulated. In situ hybridization analyses demonstrated that DdPEF-1 was expressed in both the anterior prestalk and the posterior prespore regions of the tipped aggregate, slugs and early culminants. On the other hand, DdPEF-2 was dominantly expressed in the anterior tip region of these multicellular structures. Both PEF proteins were detected as 22-23-kDa proteins in soluble fractions in the presence of EGTA but in particulate fractions in the presence of Ca(2+) by Western blotting using specific monoclonal antibodies. Together with the finding of PEF-like sequences in DNA databases of plants, fungi and protists, our results strongly suggest that Group I PEF proteins are ubiquitously present in all eukaryotes and play important roles in basic cellular functions. PMID- 11481038 TI - A serine/threonine kinase which causes apoptosis-like cell death interacts with a calcineurin B-like protein capable of binding Na(+)/H(+) exchanger. AB - We surveyed proteins capable of binding to the cytoplasmic domain of Na(+)/H(+) exchanger (NHE)1 in a rat brain cDNA library with the yeast two-hybrid system. One clone obtained coded for a protein reported previously as a human calcineurin homologous protein (CHP). Since CHP is homologous to the regulatory subunit B of calcineurin, we expected a possible interacting partner of CHP like the catalytic subunit of calcineurin (calcineurin A), and surveyed this putative partner again with the yeast two-hybrid system. A clone thus obtained coded for a kinase, which is basically the same as that reported for human DRAK2. Overexpression of the rat homologue of DRAK2 caused apoptosis-like cell death of NIH3T3 cells, which was dependent on the kinase activity, confirming the previous result for DRAK2. The purified CHP and rat DRAK2 proteins synthesized in Escherichia coli could bind in vitro. CHP and rat DRAK2 expressed in COS-7 cells were found to be localized in the Golgi apparatus and nucleus, respectively. Some of them was also found in the membrane peripheral region. When they were co-expressed in the same cells, most of CHP moved to the nucleus where rat DRAK2 is located, suggesting in vivo interaction of these proteins. However, minor but significant fractions of both proteins were also found in the membrane peripheral region. Rat DRAK2 is expressed highly in thymus, spleen, and testis, where the apoptosis plays an important role in physiology. PMID- 11481039 TI - New inhibitors of iron-containing nitrile hydratases. AB - There is growing evidence in the literature emphasizing the significance of the post-translational modification of cysteine thiols to sulfenic acids (SOH), which have been found in a number of proteins. Crystallographic and mass spectrometric evidence has shown the presence of this group in an inactive form of the industrially important enzyme nitrile hydratase (NHase). This oxidized cysteine is unique in that it forms part of the coordination sphere of the low-spin iron III at the active site of the enzyme. The presence of this unstable sulfenic group in the active form of NHase is the subject of some controversy. To try to detect this function in NHase, we have studied the inhibitory effect on nitrile hydration of reagents known to react with sulfenic acids. Two NHases were studied, namely, Rhodococcus rhodochrous R312 NHase and Comamonas testosteroni NI1 NHase, and the reagents used were meta-chlorocarbonyldicyano-phenylhydrazone (m-ClCP), 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl), and 2-nitro-5 thiocyanato-benzoic acid (NTBA). Following this approach we report three novel inhibitors of NHases. In addition, we report thiocyanate reagents that can be used to monitor NHase activity spectroscopically. PMID- 11481040 TI - Genomic organization of the human adipocyte-derived leucine aminopeptidase gene and its relationship to the placental leucine aminopeptidase/oxytocinase gene. AB - The genomic organization of the gene encoding the human adipocyte-derived leucine aminopeptidase (A-LAP) has been determined. The gene is composed of 20 exons and 19 introns and spans approximately 47 kilobases of chromosome 5q15. The gluzincin aminopeptidase motif, the HEXXH(X)(18)E zinc-binding motif essential for enzymatic activity, is encoded by exons 6 and 7. A comparison of the exon/intron boundaries, together with phylogenetic analysis, shows the close relationship between A-LAP and placental leucine aminopeptidase (P-LAP)/oxytocinase, another gluzincin aminopeptidase considered to be important for the maintenance of normal pregnancy. Primer extension analysis revealed two transcriptional initiation sites. Analysis of the sequence immediately upstream of the transcriptional initiation sites revealed that the A-LAP promoter contains no canonical TATA- or CCAAT-box, but has a PyPyA(+1)N(T/A)PyPy initiator consensus sequence and multiple putative regulatory elements. Finally, luciferase-reporter assays revealed a functional promoter activity of the 5'-flanking region of the gene, and suggested that the activity is regulated in a cell type-specific manner. PMID- 11481041 TI - The gelsolin/fragmin family protein identified in the higher plant Mimosa pudica. AB - Mimosa pudica L. rapidly closes its leaves and bends its petioles downward when mechanically stimulated. It has been suggested that the actin cytoskeleton is involved in the bending motion since both cytochalasin B and phalloidin inhibit the motion. In order to clarify the mechanism by which the actin cytoskeleton functions in the motion, we attempted to find actin-modulating proteins in the M. pudica plant by DNase I-affinity column chromatography. The EGTA-eluate from the DNase I column contained proteins with apparent molecular masses of 90- and 42 kDa. The 42-kDa band consisted of two closely migrating components: the slower migrating component was actin while the faster migrating components was a distinct protein. The eluate showed an activity to sever actin filaments and to enhance the rate of polymerization of actin, both in a Ca(2+)-dependent manner. Microsequencing of the faster migrating 42-kDa protein revealed its similarity to proteins in the gelsolin/fragmin family. Our results provide the first biochemical evidence for the presence in a higher plant of a gelsolin/fragmin family actin-modulating protein that severs actin filament in a Ca(2+)-dependent manner. PMID- 11481042 TI - A comparative study on two GNRA-tetraloop receptors: 11-nt and IC3 motifs. AB - Natural RNAs often contain terminal loops consisting of GNRA (N=A, G, C, U; R=A, G) and their receptors, which bind to the loops via long-range RNA-RNA interactions. Among several known receptors, two characteristic structural elements have been identified that are termed the 11-nt motif (CCUAAG-UAUGG) and IC3 motif (CCCUAAC-GAGGG). These two motifs that share a similar secondary structure have been shown to exhibit distinctively different binding specificities. The 11-nt motif recognizes a GAAA loop with highest specificity among the known receptors, whereas the IC3 motif distinguishes GAAA from other GNRA loops less stringently than any other receptors. To identify the elements in the receptors that determine the binding specificity, a series of chimeric receptors derived from the two motifs were prepared and their properties were examined. We identified characteristic base-pairs and a particular U residue in the receptors as such elements by means of a gel mobility shift assay that evaluates the degree of the tetraloop-receptor interaction. The relationship between the elements and the specificity is discussed together with a model that describes a possible evolutional linkage between the two receptors. PMID- 11481043 TI - A high molecular weight glutamyl endopeptidase and its endogenous inhibitors from cucumber leaves. AB - We purified a glutamyl endopeptidase that is a major foliar endopeptidase in cucumber. The endopeptidase had a molecular mass of 400 kDa, consisted of four subunits of 97 kDa, and was inactivated by SH-modifying reagents. Its optimum pH and optimum temperature were 8.0 and 30-37 degrees C, respectively. An internal amino acid sequence of the endopeptidase was highly homologous to a partial sequence of unidentified proteins deduced from genetic information for Arabidopsis thaliana, soybean and rice, but not to the sequences of bacterial glutamyl endopeptidases or animal proteases. Therefore, the unidentified proteins might be glutamyl endopeptidases and be widely distributed only among plant species. The activity of the cucumber glutamyl endopeptidase was inhibited by at least three inhibitors existing in cucumber leaves. One of the inhibitors was a competitive inhibitor of 25 kDa, which did not significantly inhibit commercial endopeptidases derived from animals and microorganisms. This suggests that the cucumber glutamyl endopeptidase might be controlled by endogenous inhibitors in vivo. PMID- 11481044 TI - Transglycosylation and reverse hydrolysis reactions of endoglycoceramidase from the jellyfish, Cyanea nozakii. AB - Endoglycoceramidase (EGCase: EC 3.2.1.123) is an enzyme capable of cleaving the glycosidic linkage between oligosaccharides and ceramides in various glycosphingolipids. We report here transglycosylation and reverse hydrolysis reactions of EGCase from the jellyfish Cynaea nozakii. Various alkyl-GM1 oligosaccharides (alkyl-II(3)NeuAcGgOse4) were synthesized when GM1 ganglioside was treated with the EGCase in the presence of 1-alkanols. Among various 1 alkanols tested, methanol was found to be the most preferential acceptor, followed by 1-hexanol and 1-pentanol. GM1 was the best donor, followed by GD1b and GT1b, when methanol was used as an acceptor. However, neither globoside nor glucosylceramide was utilized by the enzyme as a donor substrate. The enzyme transferred oligosaccharides from various glycosphingolipids to NBD-ceramide, a fluorescent ceramide, producing NBD-labeled glycosphingolipids. In addition to the transglycosylation reaction, the enzyme catalyzed the reverse hydrolysis reaction; lactose was condensed to ceramide to generate lactosylceramide in the presence of the enzyme. These results indicate that the jellyfish enzyme will facilitate the synthesis of various neoglycoconjugates and glycosphingolipids. PMID- 11481045 TI - An efficient refolding method for the preparation of recombinant human prethrombin-2 and characterization of the recombinant-derived alpha-thrombin. AB - Human recombinant prethrombin-2 was produced in Escherichia coli. The expressed prethrombin-2 formed intracellular inclusion bodies from which the protein was refolded by a simple one-step dilution process in buffer consisting of 50 mM Tris HCl, containing 20 mM CaCl(2), 500 mM NaCl, 1 mM EDTA, 600 mM arginine, 1 mM cysteine, 0.1 mM cystine, 10% (v/v) glycerol, and 0.2% (w/v) Brij-58 at pH 8.5. After refolding, prethrombin-2 was purified by hirudin-based COOH-terminal peptide affinity chromatography, and then activated with Echis carinatus snake venom prothrombin activator (ecarin). The activated protein, alpha-thrombin, was then tested for several activities including activity toward chromogenic substrate, release of fibrinopeptide A from fibrinogen, activation of protein C, and thrombin-activatable fibrinolysis inhibitor, reactivity with antithrombin, clotting activity, and platelet aggregation. The kinetic data showed no differences in activity between our recombinant alpha-thrombin and plasma-derived alpha-thrombin. The yield of refolded recombinant human prethrombin-2 was about 4 7% of the starting amount of solubilized protein. In addition, the final yield of purified refolded protein was 0.5-1%, and about 1 mg of recombinant prethrombin-2 could be isolated from 1 liter of E. coli cell culture. PMID- 11481046 TI - Duck and human pandemic influenza A viruses retain sialidase activity under low pH conditions. AB - The majority of influenza A viruses isolated from wild birds, but not humans, can replicate in the duck intestinal tract. Here we demonstrate that all duck isolates tested universally retain sialidase activities under low pH conditions independent of their neuraminidase (NA) subtypes. In contrast, the sialidase activities of most isolates from humans and pigs practically disappear below pH 4.5, with the exception of four human pandemic viruses isolated in 1957 and 1968. Sequence comparisons among duck, human, and swine N2 NA subtypes indicate that amino acids at positions 153, 253, 307, 329, 344, 347, 356, 368, 390, and 431 may be associated with the low pH stability of duck and human pandemic N2 NAs. This finding suggests that the low pH stability of duck influenza A virus NA may be a critical factor for replication in the intestinal tract through the digestive tract of ducks, and that the properties of NAs are important for understanding the epidemiology of the influenza virus. PMID- 11481047 TI - Modulation of reactive oxygen species in endothelial cells by peroxynitrite treated lipoproteins. AB - Peroxynitrite has been implicated in the oxidative modification of low-density lipoprotein (LDL) particles, and nitrotyrosine residues in the LDL have been detected in atherosclerotic plaques. Studies have suggested that lipoproteins modified by peroxynitrite lead to the onset of atherosclerotic vascular disease. We therefore prepared in vitro lipoproteins oxidatively modified by peroxynitrite (NO(2)-lipoprotein) and investigated the effect of NO(2)-lipoprotein on the viability of cultured endothelial cells. After exposure of a high-density lipoprotein (HDL) to peroxynitrite, some intermolecular complexes of apolipoproteins in HDL were detected on immunoblotting with monoclonal antibodies against apolipoprotein AI and AII, suggesting that nitration of HDL by peroxynitrite causes intermolecular cross-linking of the apolipoproteins in the particles. Treatment with 1 mM peroxynitrite increased the 3-nitrotyrosine level to 28.5 mmol/mol of tyrosine residues in the prepared NO(2)-HDL, as quantitated by HPLC, and the amount in NO(2)-lipoprotein depended on the peroxynitrite concentration. HDL exhibited a shorter lag phase and the reaction plateaued more rapidly than that with LDL. To clarify whether or not NO(2)-lipoproteins affect the function of endothelial cells, we first examined the viability of cultured human aortic endothelial cells (HAECs) exposed to NO(2)-lipoproteins. Incubation with either NO(2)-HDL or NO(2)-LDL significantly reduced the HAEC viability at 72 h. The results of RT-PCR and Western blotting showed that NO(2)-HDL markedly suppressed at 48 h not only the expressed levels of mRNA and protein but also the activity of catalase in HAECs. In contrast, NO(2)-LDL significantly reduced the expression and activity of Cu(2+),Zn(2+)-superoxide dismutase (CuZn-SOD) in the cells. Neither NO(2)-HDL nor NO(2)-LDL interfered with nitric oxide production or expression of cyclooxygenases and NADPH oxidase in HAECs. Increased radical production in NO(2)-lipoprotein-treated HAECs implied that reactive oxygen species such as superoxide anions and hydroxyl radicals may contribute to the mechanism of the toxic effect induced in endothelial cells by NO(2)-lipoprotein. Overall, NO(2)-lipoprotein may lead to deterioration of the vascular function through these endothelial cell responses. PMID- 11481048 TI - In vitro refolding of porcine pepsin immobilized on agarose beads. AB - Since in vitro refolding of pepsin has long been attempted without success, it has been suspected that pepsin has no intrinsic refolding ability. In the present study, in order to eliminate unfavorable intermolecular interactions bringing about aggregation and autoproteolysis, we immobilized pepsin onto agarose beads. This technique enabled us to search extensively for appropriate refolding conditions without limitation of the refolding period. Renaturation of immobilized pepsin was observed exclusively at pH 3-5. This process was extremely slow and reached equilibrium after 300 h. Sixty percent of the proteolytic activity was recovered at pH 5. Addition of salts raised the recovery to 80% but had no significant effect on the refolding rate, suggesting that the salts mainly stabilize the native state of pepsin. This is the first report on the successful in vitro refolding of pepsin. PMID- 11481049 TI - Cloning and expression of human anti-tumor necrosis factor-alpha monoclonal antibodies from Epstein-Barr virus transformed oligoclonal libraries. AB - Peripheral blood was obtained from a healthy human volunteer and transformed with Epstein-Barr virus (EBV). This produced an oligoclonal cell library in culture medium that was screened by ELISA for anti-human tumor necrosis factor-alpha (TNFalpha) activity. RNA from two positive clones was applied to RT-PCR using antibody-specific primers, and the light (kappa and lambda) and heavy chain genes (gamma and mu) were cloned into the plasmid vector pFab1-His2. The antibodies produced in Escherichia coli as Fab fragments were assayed for anti-TNFalpha activity utilizing ELISA. Two IgG1/kappa anti-TNFalpha antibodies and two IgM/kappa anti-TNFalpha antibodies were isolated. DNA sequence analysis showed that the VL and VH gene families of IgM and IgG were the same. Both the antibodies showed almost the same activity on ELISA-testing. Ten clones randomly selected from light (kappa and lambda) and heavy (gamma and mu) chain genes in the oligoclonal cell library 1D5 were sequenced, and each gene (kappa, lambda, gamma, and mu) was found to be composed of one to three different genes. These data support the conclusion that the cell clone is oligoclonal at the molecular level. PMID- 11481050 TI - A growth signal with an artificially induced erythropoietin receptor-gp130 cytoplasmic domain heterodimer. AB - We report a strategy for generating efficient signal transduction with unnatural heterologous receptor combinations. As previously described [Ueda, H., Kawahara, M. et al. (2000) J. Immunol. Methods 241, 159-170], chimeric receptors composed of the V(H)/V(L) domains of anti-hen egg lysozyme antibody HyHEL-10 and N terminally truncated erythropoietin receptor (EpoR) can be activated by lysozyme. When the cytoplasmic domains of these receptors were substituted with one derived from gp130, IL-3 dependent Ba/F3 cells expressing both V(H)-gp130 and V(L)-gp130 grew dose-dependently when given lysozyme without IL-3. However, cells expressing the heterologous pair of V(H)-gp130 and V(L)-EpoR also showed more efficient and stricter lysozyme-dependent proliferation in the absence of IL-3, indicating this combination is as an efficient and strict signal transducer as wild-type EpoR. The immunoprecipitation data indicated the existence of a preformed V(H)-gp130 and V(L)-EpoR heterodimer in the absence of lysozyme, suggesting the crucial role of a receptor conformational change in signal triggering as well as wild-type EpoR and gp130. Phosphorylation of JAK2, STAT3, and STAT5 was observed upon the addition of lysozyme, suggesting the activation of both EpoR- and gp130-derived signals. PMID- 11481051 TI - Identification and characterization of an antibacterial peptide of the 26-kDa protease of Sarcophaga peregrina with antibacterial activity. AB - Previously, we purified a serine protease with a molecular mass of 26 kDa that exhibits potent antibacterial activity from a pupal extract of Sarcophaga peregrina (flesh fly). We divided this protease into 12 peptides and examined their antibacterial activity. A peptide corresponding to residues 155 to 174 (peptide 9) was found to exhibit antibacterial activity comparable to that of the 26-kDa protease. When Escherichia coli was treated with peptide 9, the permeability of both the outer and inner membranes increased, and substrates for beta-lactamase and beta-galactosidase entered the cells, but beta-galactosidase did not leak out of the cells under these conditions. It was suggested that residues 6 to 18 of peptide 9 form an amphiphilic alpha-helix under hydrophobic conditions with an N-terminal basic loop and then interact with acidic phospholipids in the bacterial membranes. PMID- 11481052 TI - [Transbronchoscopic ultrasonography: a debatable procedure]. PMID- 11481053 TI - [How long should a patient stay in the hospital after lung resection?]. AB - OBJECTIVE: To describe the hospital stay of patients undergoing lobectomy or pneumonectomy in comparison with reference data from the Spanish National Health Service. MATERIAL AND METHOD: Prospective study of all consecutive lobectomy or pneumonectomy cases from January 1998 through December 2000. Data collected prospectively were as follows: date of birth, main diagnosis, dates of admission and discharge, surgical procedure, complications and postoperative exitus. Data collected retrospectively included all information related to readmission over the 30 days following discharge. Reference data were obtained from the web page of the Spanish Ministry of Health and Consumer Affairs. RESULTS: Mean hospital stay for the 279 patients studied was 8.3 days (8.4 for 214 lobectomy patients and 7.7 for 65 pneumonectomy patients). For either of the procedures the 50 percentile was 7 days. The rate of readmission was 8.6%. The 83 patients with postoperative complications required a mean stay of 11.9 days (p < 0.001). The reference data for 1998 were 17.5 days for a lobectomy (n = 1,443) and 19.8 days (n = 693) for pneumonectomy. CONCLUSION: The mean hospital stay of reference is far longer than that which we consider adequate for pulmonary resection. PMID- 11481054 TI - [Monitoring domiciliary oxygen therapy from a district hospital]. AB - OBJECTIVES: This study aimed a) to evaluate correct use of domiciliary oxygen therapy (DOT); b) to estimate the prevalence of DOT, and c) to evaluate DOT based on the same parameters after intervention by the monitoring team. PATIENTS AND METHOD: Cross-sectional, prospective study of all patients receiving DOT before and after initiation of monitoring. We administered spirometric tests, analyzed indications for and compliance with DOT and monitored pulse oxymetry in order to adjust oxygen flow. RESULTS: Seventy-six patients were receiving DOT (63/100,000 inhabitants). Among the 60 patients with COPD, half met ideal indications for prescribing DOT, 65% complied with over 15 h of DOT, and hypoxemia was not corrected for 26%. Monitoring resulted in withdrawal of DOT from 28 patients (reduction of 37%), and DOT was prescribed for 27 new patients, 11 of whom received liquid oxygen. At the end of the study, 46 patients were receiving DOT (38/100,000 inhabitants). CONCLUSIONS: a) Ideal indications for DOT, adequate compliance and correction of hypoxemia were observed in 54% of the patients undergoing therapy; b) creation of a special service to care for patients receiving DOT improves monitoring, and c) the prevalence of DOT in our area has been reduced from 63 to 38/100,000 inhabitants. PMID- 11481055 TI - [Anti-smoking specialist medical prescription or medical counseling in primary care: opinions from the other side of the table]. AB - OBJECTIVES: To estimate the frequency, characteristics and follow-up of anti smoking counseling given to patients using a primary care facility; to survey patient opinion of anti-smoking advice offered. MATERIAL AND METHODS: Cross sectional, descriptive study at a primary care facility. SUBJECTS: random sample of 330 patients between 15 and 75 years of age using the primary care facility in March and April 2000. MEASUREMENTS: confidential survey by self-administered questionnaire on smoking, chronic disease related to smoking, anti-smoking advice given and patient opinion of anti-smoking counseling by health care givers. RESULTS: The prevalence of smoking was 31.9% (100). Among patients who were either smokers or ex-smokers (162), advice to stop smoking had been received from the primary care physician by 47 patients (29.2%), from a specialist by 19 (11.8%), from a nurse by 14 (8.7%), from a pharmacist by 5 (3.1%), and from family by 34 (21.1%). We found no statistically significant differences related to gender or age for advice to stop smoking, although the presence or absence of chronic disease did affect counseling. Fifty-four (65%) of those who had been advised to quit considered that the time spent counseling was adequate. Ten patients (12%) received an informative brochure in addition to counseling. Forty smokers (40%) stated that they would quit smoking if their doctor advised them to and 45 (45%) would do so if they had a smoking-related disease. Thirty-three smokers (33%) believed that they should receive advice about smoking at all visits to the doctor, whereas 38 (38%) believed such counseling was appropriate only if the visit was for a smoking-related disease. CONCLUSIONS: The prevalence of smoking among patients who use primary care facilities is high. The frequency of advice to quit smoking is low, and reminders and follow-up are minimal. After learning a patient's habits and studying them, professional anti-smoking advice should always be given, with appropriate monitoring and follow-up. PMID- 11481056 TI - [Cardiopulmonary exercise tests]. PMID- 11481057 TI - [Recommendations for the care of the patient with chronic obstructive pulmonary disease]. PMID- 11481058 TI - [Strategies for muscular training in chronic obstructive pulmonary disease. Training of resistance, strength, of both?]. PMID- 11481059 TI - [Acute interstitial pneumonia: favorable outcome with steroid therapy]. AB - We report the case of a 65-year-old man with no history of respiratory disease who came to the emergency room complaining of fever, cough, yellowish sputum and increasing dyspnea of one week's duration. Severe respiratory insufficiency was evident upon examination and a chest film showed diffuse pulmonary infiltrates. Microbiological tests were negative. Lung biopsy gave evidence consistent with diffuse alveolar damage in organizing phase. Evolution of symptoms and blood gases was good after corticosteroid treatment was begun, although pulmonary fibrosis was still evident in a follow-up radiograph. PMID- 11481060 TI - [Massive hemoptysis secondary to cardiac insufficiency in a context of dilated myocardiopathy]. AB - Massive hemoptysis is a life-threatening emergency that rarely occurs in association with heart failure and is even less frequent when the underlying cause is dilated myocardiopathy. We report the case of a 69-year-old man with undiagnosed heart disease who presented with severe hemoptysis. Images showed evident cardiac insufficiency. The unusualness of the case obliged us to rule out other diseases through differential diagnosis before starting to treat the underlying condition. PMID- 11481061 TI - [Spirometry in primary care?]. PMID- 11481063 TI - [Pleural effusion with alternating presentation nad tuberculous spondylodiscitis]. PMID- 11481064 TI - ["Trapped lung" syndrome: a cause of recurrent pleural effusion]. PMID- 11481065 TI - [Increased bile lithogenicity after simultaneous total parenteral nutrition and octrotide. A model of calcium palmitate Gallstones]. AB - Gallbladder stasis and gallstone formation are well-known complications of both fasting-associated total parenteral nutrition (TPN) and long-term treatment with octreotide. Additive noxious effects to hepatobiliary function may develop when both therapies are given together as treatment of enteric fistulae. The aim of this study was to assess the risk of gallstone formation during treatment with TPN and octreotide separately and in combination. We studied four groups of New Zealand rabbits (n = 32) during a 2-week intervention period: 14 chow-fed controls; 6 fasted and TPN-fed; 6 chow-fed and administered octreotide, and 6 fasted and treated with both TPN and octreotide. After treatment, the bile duct was cannulated and the gallbladder and liver tissue were obtained for histological analysis. Hepatic and gallbladder bile were retrieved for microscopic examination and measurement of biliary lipids, bilirubin, calcium, total protein, and cholesterol nucleation time. The chemical composition of gallstones was also analyzed. The results of the study suggest that simultaneous administration of these two therapies in rabbits has additive effects on gallbladder stasis and bile lithogenicity. The administration of both treatments can play an important role in the formation of calcium palmitate gallstones in these animals. PMID- 11481066 TI - [Role of early colonoscopy in severe acute lower gastrointestinal bleeding]. AB - BACKGROUND: Severe acute lower gastrointestinal bleeding (SALGIB) accounts for 15% of cases of acute lower gastrointestinal bleeding (ALGIB). The incidence increases with age and comorbidity. Identification of the origin of bleeding may be difficult. Colonoscopy has been proposed as the primary investigative tool. AIM: To assess the role of early colonoscopy as the primary method of evaluation in patients with SALGIB. PATIENTS AND METHOD: Retrospective study based on a guideline for clinical practice approved in our institution. The study included 50 patients with SALGIB admitted to our gastrointestinal bleeding unit between January 1998 and April 2000. SALGIB was suspected when patients fulfilled two or more of the following criteria: 1) significant hemodynamic compromise, 2) decrease in hemoglobin 2 g/dl, and 3) transfusion requirement >= 2 blood units. Early colonoscopy was performed within 24 hours of onset of bleeding. An accurate endoscopic diagnosis was established if a lesion with active bleeding, visible non-hemorrhagic vessel or adherent red clot was identified. A presumptive diagnosis was made when hematochezia or fresh blood localized in a colonic segment, associated with a single, potentially hemorrhagic lesion, was observed and when the results of esophagogastroduodenoscopy were negative. Colonoscopy, esophagogastroduodenoscopy, barium studies, nuclear scan and angiography were performed. RESULTS: Two hundred twenty-two patients were admitted for ALGIB. Fifty patients(22%) fulfilled the SALGIB criteria. The male/female ratio was 1:1. Definitive diagnosis was accurate in 20 patients. The most frequent cause was angiodysplasia (6 patients) and rectal ulcer (6 patients). Eighteen patients had a presumptive diagnosis; of these 14 had diverticulosis. In 12 patients, no cause was identified. Colonoscopy was performed in 45 patients, of which 32 were performed early and 13 electively. Accurate endoscopic diagnosis was more frequently established with early colonoscopy than with elective colonoscopy (15 [47%] vs 2 [15%], p < 0.05). The results of urgent nuclear scans contributed to accurate diagnosis in 5 out of the 10 patients in whom this technique was performed. Angiography was performed in 2 patients. Endoscopic therapy was attempted in 4 patients, all during early colonoscopy. Ten patients (20%) underwent surgery and 3 patients (6%) died. CONCLUSIONS: In 22% of patients with ALGIB admitted to our hospital bleeding was severe. Colonoscopy is the diagnostic tool of choice. When performed within 24 hours of hospital admission, this technique provides more accurate diagnosis than when performed electively. PMID- 11481067 TI - [Clinical and functional results after Laparoscopic fundoplication. Prospective evaluation]. AB - OBJECTIVE: To prospectively assess the medium- and long-term clinical and functional results in consecutive patients undergoing laparoscopic procedures for gastroesophageal reflux. PATIENTS AND METHOD: The series included the interventions performed or assisted by the first author in 325 patients referred for surgical evaluation between December 1992 and June 2000. Forty patients were excluded from the study because they did not fulfil the inclusion criteria for laparoscopic surgery. A further 20 patients who underwent laparoscopic surgery were excluded because they required conversion to open surgery. Thus, 265 patients were included in the study. One hundred ninety-six total fundoplications (Nissen-Rossetti) and 69 partial fundoplications(42 posterior-Toupet and 27 anterior-Dor) were performed. The surgical technique employed was a slight modification of that described by Dallemagne et al with five-trocars. Preoperative evaluation was established by clinical features, endoscopy with biopsy, barium contrast radiography, esophageal manometry and esophageal pH monitoring. RESULTS: The mean follow-up was 47.1 3.3 months (range: 4-92 months). Heartburn, regurgitation, and dysphagia were relieved in 92.5%, 97.7%, and 88.1% of patients respectively. Complete relief of symptoms was achieved in all patients who presented preoperative respiratory symptoms as a complication of gastroesophageal reflux. Esophagitis healed in 98.2% of patients with preoperative esophagitis. Correction of lower esophageal sphincter pressure and lower esophageal sphincter length were statistically significant compared with preoperative status (p = 0.006 and p = 0.003, respectively). Pre- and postoperative differences in the percentage of patients with a pH < 4 in 24-hour ambulatory esophageal pH monitoring were also significant (p = 0.005), confirming correction of acid reflux. Morbidity appeared in 6.7% of patients and there was no mortality. Conversion to open procedures was required in 7.5%. The mean operative time was 115 6.3 minutes. CONCLUSIONS: These preliminary results suggest that when performed by experienced surgeons laparoscopic fundoplication provides an excellent alternative in selected patients with gastroesophageal reflux. Both medium- and long-term clinical and functional results, as well as morbidity, were satisfactory and were similar to those of open fundoplication. PMID- 11481068 TI - [Atypical colonic stromal tumor]. AB - We present the case of a female patient with atypical gastrointestinal stromal tumor. The young woman presented asymptomatic colonic perforation and showed atypical immunohistochemical findings. The various clinicopathologic characteristics and diagnostic tests, as well as treatment and distinct behavior of these tumors, are discussed. PMID- 11481069 TI - [Lower gastrointestinal bleeding due to Dieulafoy's in the upper colon]. AB - Dieulafoy's lesion is a vascular anomaly generally located in the proximal stomach, although it has also been documented in other areas such as the colorectum. It is mainly found in men aged between 50 and 70 years, and represents less than 2% of acute gastrointestinal hemorrhagic episodes.A 66-year old woman who was undergoing oral Diclofenac treatment presented with black stools. Endoscopy revealed acute duodenal erosions with no signs of bleeding. Black stools persisted after Diclofenac was discontinued and omeprazole treatment was started and the patient was admitted to hospital after 7 days. Colonoscopy revealed active bleeding in the upper colon, which ceased after sclerosis with ethanolamine oleate. The patient was discharged from hospital but was readmitted 10 days later because of rebleeding. The results of upper endoscopy were normal and colonoscopy performed 3 days later detected neither lesions nor bleeding. The pathogenesis of Dieulafoy's lesion is not well known, although it could be caused by erosion of the mucous lining of a vessel. Definitive diagnosis is histologic, although certain endoscopic diagnostic signs have been described. Endoscopic diagnosis is sometimes difficult; in such cases, arteriography should be employed, both for diagnostic and therapeutic purposes. The treatment of choice is endoscopic and the use of two hemostatic methods is advisable. If these procedures fail, surgery is required. Arteriography is the most suitable alternative in patients who are poor candidates for surgery. PMID- 11481070 TI - [Acute peritonitis due to perforating crohn's disease]. AB - Peritonitis is an infrequent complication of perforating Crohn's disease. Three patients, aged between 25 and 41 years, underwent surgery for peritonitis due to perforating Crohn's disease in the terminal ileum. The underlying disease differed in each patient (one patient was asymptomatic and two presented acute episodes of the disease; of these two patients, corticoids were required in one to control symptomatology). Treatment consisted of intestinal resection with primary anastomosis. In all three patients evolution was satisfactory. Taking only patients treated in our hospital into account, the frequency of peritonitis due to perforating Crohn's disease in our area is high (more than 11%). Intestinal resection with primary anastomosis should be performed whenever possible. PMID- 11481071 TI - [Aspirin and nitroglycerine: a good combination for gastric mucosa]. PMID- 11481072 TI - [New perspectives in the treatment of chronic hepatitis C]. PMID- 11481073 TI - [Prognostic factors for acute pancreatitis]. PMID- 11481074 TI - [Bilateral Krukenberg's tumor secondary to gastric cancer]. PMID- 11481075 TI - [Paracetamol-codeine induced hepatic colic]. PMID- 11481076 TI - [Commentary on the letter: "Azathioprine and cavum carcinoma in a male patient with autoimmune hepatitis and Crohn's disease"]. PMID- 11481077 TI - [Amoxycillin-clavulanic acid in patients with spontaneous bacterial peritonitis]. PMID- 11481078 TI - [Gastric ulcer and intestinal occlusion: which came first - Helicobacter pylori infection ot the bezoars]. PMID- 11481079 TI - [Hyperamylasemia due to macroamylasemia as the first manifestation of celiac disease]. PMID- 11481082 TI - [Clinical and pathological characteristics and clinical course of patients with breast cancer and BRCA1/BRCA2 mutations]. AB - BACKGROUND: Clinico-pathological differences between BRCA1 or BRCA2 mutation associated breast cancer (BC) and sporadic BC are little known. PATIENT AND METHODS: We analysed the clinico-pathological characteristics and clinical follow up of 30 patients with BC. BRCA1 and BRCA2 mutations were detected by SSCP and PTT. RESULTS: There were no differences in age, size or nodal status at the time of diagnosis. Mammography features were more heterogeneous in BRCA2 than in BRCA1 BC. All BRCA1 mutation-associated BC corresponded to infiltrating ductal carcinomas (20% medullary carcinomas) with a more aggressive pathological behavior. The frequency of local recurrences was 14% in BRCA1 and 20% in BRCA2. Contralateral BC and ovarian cancer (OC) were observed in 27% and 20% of BRCA1 cases, respectively, and 6% and 6% of BRCA2 cases. The median follow-up in BRCA1 and BRCA2 BC was 131 and 54 months, respectively. CONCLUSIONS: There were no differences in age at diagnosis and stage between BRCA1 and BRCA2 breast cancer. The mammographic pattern in BRCA2 was more heterogeneous. BRCA1 mutations were associated with more aggressive histopathologic findings and a higher risk of a second BC and OC. PMID- 11481083 TI - [Assessment of paclitaxel treatment of non-small-cell lung cancer]. AB - BACKGROUND: Our goal was to assess the effectiveness of paclitaxel therapy in patients with non-small-cell lung cancer (NSCLC) by means of a meta-analysis of published clinical trials. MATERIAL AND METHOD: We carried out a search of controlled and randomized clinical trials which evaluated treatment with paclitaxel in patients with NSCLC from January 1996 to April 2001, regardless of any other associated therapy and without restrictions in the publication language. We also performed a sensitivity analysis and an analysis of sample heterogeneity. RESULTS: Six randomized and controlled studies fulfilled the inclusion criteria. Results from the analysis of effectiveness favoured significantly treatment with paclitaxel (OR 95% total responders: 1.42 [1.16 1.74]; p = 0.07). These results remained unchanged with the sensitivity analysis. Analysis of survival after 1 year of treatment was not significant (OR 95% 0.96 [0.79-1.17]; p = 0.2) CONCLUSIONS: Paclitaxel in patients with NSCLC offers a therapeutic advantage over other chemotherapy regimes with an overall OR of 1.42. However, this therapy does not appear to offer a significant survival advantage after 1 year. PMID- 11481084 TI - [Weight gain along adult life]. AB - BACKGROUND: Obesity is a serious health problem with a rather difficult treatment and prevention. Estimating weight gain along adult life may be useful for preventing adult obesity. SUBJECTS AND METHOD: This cross-sectional study included a representative sample of 1,286 adults who self-reported a weight gain from age 20 years. RESULTS: The average weight gain per decade was about 4.5 kg in males and females aged 20 to 49 years. Among subjects aged 50 to 69 years, it was 1 kg in men and 2 kg in women, approximately. Body weight decreased from age 70 years onwards. CONCLUSION: Weight gain appears to be progressive from the third up to the seventh decade of life in both sexes, yet it is more persistent in women. PMID- 11481085 TI - [Hepatectomy: indications, techniques and results]. PMID- 11481086 TI - [Treatment of systemic necrotizing vasculitis and giant cell arteritis]. PMID- 11481087 TI - [Hyperprolactinemia coexisting with hypophyseal adenoma or microadenoma]. PMID- 11481088 TI - [Pneumococci or "neumococos"? On the CDC and the authority sophism]. PMID- 11481089 TI - [Imipenem/cilastatin-induced acute thrombocytopenia]. PMID- 11481090 TI - [Hypertensive crisis and transitory left brunch block with QT interval prolongation associated to moxifloxacin]. PMID- 11481092 TI - [Retroperitoneal angiomyolipoma and pregnancy]. PMID- 11481093 TI - [Relationship between T-cells subsets and prognostic markers in HIV-1-infected children]. AB - BACKGROUND: To investigate the relationship between peripheral blood T-cell subsets and both CD4+ T-cell percentage and viral load (VL) in HIV-1-infected children. PATIENTS AND METHOD: We studied 50 HIV-1-infected children on antiretroviral therapy. T-cell subsets were determined by flow cytometry. The VL was quantified using standardized molecular methods. RESULTS: Memory (CD45RO+), activated memory (CD45RO+HLA-DR+) and CD45RA-CD62L+ (memory cells expressing L selectin) CD4+ and CD8+ T-cells correlated positively with the VL and negatively with the percentage of CD4+ T-cells. Inversely, naive CD4+ and CD8+ T-cells (CD45RA+CD62L+) correlated positively with the percentage of CD4+ T-cells and negatively with the VL. HLA-DR+, CD38+ or HLA-DR+CD38+CD4+ and CD8+ T-cells correlated also positively with the VL and negatively with the percentage of CD4+ T-cells (with the exception of CD4+CD38+ which did not show any association with the VL). CD8+CD28+ T-cells correlated positively with the percentage of CD4+ T cells and negatively with the VL, whereas CD8+ CD57+ and CD8+CD28-CD57+ exhibited an opposite association. CONCLUSIONS: Our data suggest a relationship between the different lymphocyte subsets (memory, naive, activated and effector T-cells) and the most commonly used markers in clinical practice, namely the viral load and the CD4+ T-cell percentage. Some of these subsets may be useful to determine the virologic and immunologic status in HIV-1-infected children. PMID- 11481094 TI - [Compliance of hormone replacement therapy in menopausal women]. AB - BACKGROUND: The effectiveness of a given treatment is only achieved with a due compliance. Our objective was to know the compliance degree of hormone replacement therapy (HRT) in menopausal women. METHOD: A descriptive observational study was carried out in three hospitals in the Valencia Community (Spain). We included menopausal patients who were visited in a menopause unit from 1989 to 1999 and who were administered HRT. We analyzed age, age at menopause, type of menopause, age at starting HRT, education level, prescription reason, treatment duration, information level, side effects, and causes leading to withdrawal. RESULTS: The study was performed in 363 women. There were 75% probabilities that women fulfilled the therapy for 5 years. The median of fulfillment was 11 years. Those women who had side effects were more prone to withdraw the treatment. By contrast, women who experienced benefits were less prone to withdraw it. Risk of withdrawal was also lower in cases of postsurgery menopause and in women who were younger than 55 years when they started HRT. CONCLUSIONS: In our study, the compliance level was high and it was determined either by treatment benefits or side effects, type of menopause and age at starting HRT. PMID- 11481095 TI - [Effects of levofolinic acid on plasma homocysteine concentrations in healthy and young women in preconceptional care]. AB - BACKGROUND: Increases in total plasmatic homocysteine (tHcy) represents a risk factor for neural tube defects. We studied the effects of levofolinic acid (l,5 formyl-tetrahydrofolic) on the plasmatic tHcylevels in women of child-bearing age. MATERIALS AND METHOD: Healthy women aged 18-35 years (n = 30) received levofolinic acid, 5 mg/day,orally for 30 days. Both tHcy and intraerythrocytic folate levels were measured before treatment (day 0), on days 2, 5, 10 and 30 within the treatment period and on days 30 (day 60) and 60 (day 90) after the treatment was finished. Plasmatic tHcy was measured by fluorescence polarisation immunoassay and intraerythrocyticfolates by chemiluminescent immunoassay. RESULTS: Plasmatic tHcy decreased from the second day of treatment onwards (day 0 vs. 2: mean of difference: -1.24 micromol/l; CI 95% = -0.84 to -1.63; p < 0.001). The maximum decline (32.3%) was observed after 30 days (mean of difference = 2.72 micromol/l; CI 95% = -2.20 to -3.24; p < 0.001).After finishing the treatment, the hypohomocysteinic effect persisted up to days 60 (mean of difference = -2.67 micromol/l; CI 95% = -2.07 to -3.26; p < 0.001)and 90 (mean of difference = -1.49 micromol/l; CI 95% = -0.94 to -2.03; p < 0.001). The response was greater when the plasmatic tHcy concentration was >= 9 micromol/l. CONCLUSIONS: Levofolinic acid leads to a earlier, intense and persistent drop of the plasmatictHcy levels. PMID- 11481096 TI - [Is it possible to eradicate vertical transmission of HIV infection?]. PMID- 11481097 TI - [New strategies to determine hematopoietic progenitors by flow cytometry]. PMID- 11481098 TI - [Clinical management: why and what for?]. PMID- 11481099 TI - [Bleeding peptic ulcer. Can the prognosis be accurately estimated and the hospitalization prevented?]. PMID- 11481100 TI - [Nebulized antibiotics in patients with cystic fibrosis]. PMID- 11481101 TI - [Angina pectoris induced by oral dipyridamol]. PMID- 11481102 TI - [Acute posterior multifocal placoid epitheliopathy and antineutrophil cytoplasmic antibodies. A frequent association?]. PMID- 11481103 TI - [Sinus arrest after the administration of intravenous metoclopramide]. PMID- 11481104 TI - [Sudden death attributed to ventricular arrhythmia induced by an over-the-counter cold medicine]. PMID- 11481106 TI - [Santiago Ramon y Cajal and cardiology: His little known discovery of sarcolemma in the cardiomyocytes]. AB - The first description of sarcolemma in cardiomyocytes was reported by Ramon y Cajal in Textura de la fibra muscular del corazon, published in 1888. In this article, he summarized his observations of the structure of cardiac fibres applying gold chloride dyes and posterior corrosion by acids. The sarcolemma, the fundamental membrane of the cardiac muscle cell, which was not recognized at that time by other researchers, was demonstrated by Ramon y Cajal who provided the first evidence supporting its existence and hypothetized about its functional significance. PMID- 11481107 TI - [The right ventricle revisited]. PMID- 11481108 TI - [Evaluation of the safety of stress echocardiography in Spain and Portugal]. AB - INTRODUCTION. There are few studies that evaluate the safety of stress echocardiography with discordant results. They are done in well-trained centers with highly selected populations leading to selection bias. OBJECTIVE: To assess the safety of the different modalities of stress echocardiography in Spain and Portugal. METHOD: Severe complications were retrospectively analyzed during the stress echocardiography performed in 29 hospitals in Portugal and Spain, from the beginning of their activity to September, 1999. In this period 22,105 stress echocardiograms were performed: 10,975 exercise echos, 2,969 low dose dobutamine echos, 6,832 high dose dobutamine echocardiograms, 1,276 dypiridamole echocardiograms, 41 paced echocardiograms and 12 with adenosine. A complication was defined as severe when it was life-threatening or led to hospital admission. RESULTS: We registered 26 complications, one death, 3 ventricular fibrillations, 10 sustained ventricular tachycardias, 2 complete atrioventricular blocks, 6 acute myocardial infarctions, 2 ruptures of the free wall or ventricular septal defects, 1 transient ischemic attack and 1 severe symptomatic hypotension. We had one severe complication for every 2,743 exercise stress, 1 every 1,231 dypiridamole, 1 every 325 high dose dobutamine without any complications with low dose dobutamine stress. We found a relationship between experience in dobutamine stress echocardiography and the frequency of complications. Three complications appeared once the test was finished. CONCLUSIONS: The stress echocardiography is a safe technique, but not harmless. The exercise stress echo is the safest of all the modalties of stress echocardiography. There is a relation between experience and the number of complications. PMID- 11481109 TI - [Prognostic significance of the implantation of a temporary pacemaker in patients with acute myocardial infarction]. AB - OBJECTIVE: Indication of temporary pacemakers in patients during acute myocardial infarction was widely studied in the pre-thrombolytic era without having determined whether the generalization of fibrinolysis might have changed the overall incidence and significance of temporary pacemakers. Our aim was to determine the incidence and the prognostic significance of insertion of temporary pacemakers in patients with acute myocardial infarction. PATIENTS AND METHODS: In a study involving 1,239 patients consecutively admitted to hospital with acute myocardial infarction we studied clinical characteristics and prognosis depending on temporary pacemaker insertion or not. We performed an univariate analysis on in-hospital mortality and those selected variables were introduced in to a logistic regression analysis. RESULTS: A temporary pacemaker was indicated in 55 patients (4.4%), prophylactically in 22% and therapeutically in 78%. Temporary pacemakers were inserted in 55% of the patients with advanced AV block and in the 10% of the patients with bundle-branch block. Pacemaker insertion was associated with higher number of affected leads in the ECG, and higher CK peak, regardless of the association with thrombolysis. The following complications were more often observed in patients with temporary pacemakers: atrial fibrillation, heart failure, right bundle-branch block, advanced atrioventricular block and in hospital mortality (45.4 vs 10.2%; p < 0.001). Need for a temporary pacemaker was less frequent in patients treated with thrombolytics compared with those not treated (3.0 vs 6.1%; p < 0.02). Pacemaker insertion had an independent value for predicting in-hospital mortality (OR = 5.51; 95% CI, 2.71-11.19). CONCLUSION: The insertion of a temporary pacemaker in acute myocardial infarction is less frequent nowadays than on the pre-thrombolytic era. Pacemaker insertion is associated with higher indices of infarct extension and in-hospital mortality, having independent prognostic value on the in-hospital mortality. PMID- 11481110 TI - [Independent predictive factors of acute and first year success after electrical cardioversion in patients with chronic atrial fibrillation]. AB - OBJECTIVES: We retrospectively analyzed the predictive factors of successfully electrical cardioversion in patients with chronic atrial fibrillation. METHODS: We included 118 patients, 68 men and 50 women, with a mean age of 65.1 years and a length of arrhythmia evolution of 83.3 days. These patients consecutively underwent electrical cardioversion in our Cardiology Department with a follow-up of one year to determine relapses. Structural cardiopathy was observed in 63.6% of the patients and 43.7% presented a left atrium between 4 and 5 cms. We analyzed the clinical and echocardiographic factors which predict the acute and first year success of electrical cardioversion. RESULTS: The cardioversion was effective in 73.7% (CI 95%, 64.6%-81.1%) of the patients and 35.6% (CI 95%, 25.8% 46.6%) had a relapse within the first year. The inexistence of cardiomyopathy and therapy with amiodarone were predictive of acute success (p < 0.04 and p < 0.03, respectively). The length of arrhythmia evolution did not predict acute success but did so when relapses were analyzed. The size of the left atrium is predictive of both acute and long term success (p < 0.02 and p < 0.001, respectively). Logistic regression showed that the size of the left atrium and the patient's age were the only predictive factors of acute and first year success. CONCLUSIONS: Electrical cardioversion is very efficient in the short-term, despite numerous relapses. Patient age and the size of left atrium are associated with acute and long-term success of cardioversion. PMID- 11481111 TI - [Usefulness of superficial electrocardiogram in the diagnosis of narrow QRS tachycardias: correlation with intracavitary electrocardiograms]. AB - INTRODUCTION AND OBJECTIVES: The electrocardiogram is an important tool to diagnose tachycardias. The sequence of analysis of the superficial electrocardiogram was evaluated for the diagnosis of supraventricular tachycardias with narrow QRS complexes (<= 110 ms) establishing correlation with intracavitary electrograms. PATIENTS AND METHODS: Between November 1996-October 1998, 674 electrophysiological studies were revised. 173 cases were selected, during which supraventricular tachycardias were inducted. In each study, the superficial electrocardiographic leads were correlated with the intracavitary electrograms during tachyarrhythmias. The P waves were located with respect to the R waves, the R-T or T-R' intervals, and the T waves, and relations between atrioventricular and ventriculoatrial intervals were established with the P-R' and R-P intervals, respectively, in the electrocardiogram. RESULTS: 107 patients were female. Upon locating the P waves in the electrocardiogram, in each supraventricular tachycardia we observed: a) P waves coincided with the R waves: 30 typical nodal reentries; b) P waves between R and T waves: 95 orthodromic atrioventricular reentries, 27 nodal reentries, 1 atrial tachycardia; c) P waves between T and R' waves: 4 nodal reentries, 5 orthodromic atrioventricular reentries, 7 atrial tachycardias, and T transition at position -511) in patients with schizophrenia. METHOD: In vivo magnetic resonance imaging and genotype analysis were used in the examination of 44 male schizophrenic patients and 48 healthy male comparison subjects. RESULTS: No association between the interleukin-1beta polymorphism and schizophrenia was detected. Within the patient group, bifrontal-temporal gray matter volume deficits and generalized white matter tissue deficits in allele 2 carriers (genotype T/T or C/T) were found. In contrast, the interleukin-1beta polymorphism had no influence on brain morphology within the healthy subjects. CONCLUSIONS: The data suggest that allele 2 within the promoter region of the interleukin-1beta gene at position -511 contributes to structural brain alterations in patients with schizophrenia. PMID- 11481170 TI - Sensory gating deficit expressed by a disturbed suppression of the P50 event related potential in patients with Alzheimer's disease. AB - OBJECTIVE: Disturbed sensory gating has been related to attention deficit and greater distractibility in patients with schizophrenia, and dysfunction of the alpha-7 subunit of the cholinergic nicotinic receptor has been discussed as its biological basis. Alzheimer's disease is characterized by a cholinergic deficit, and postmortem studies have reported alpha-7 receptor loss in patients with Alzheimer's disease. In this study, the authors tested whether sensory gating is disturbed in patients with Alzheimer's disease. METHOD: Suppression of the P50 event-related potential following the second click of a double-click paradigm, a measure of sensory gating, was assessed in 17 Alzheimer's disease patients and 17 comparison subjects. RESULTS: Alzheimer's disease patients showed less P50 suppression following the second click relative to the comparison subjects. CONCLUSIONS: Disturbed sensory gating might result from cholinergic dysfunction and possibly from alpha-7 nicotinic receptor loss in patients with Alzheimer's disease. Prospective studies should investigate the relationship between sensory gating deficit and behavioral disturbances in Alzheimer's disease patients. PMID- 11481171 TI - Hippocampal and anterior cingulate activation deficits in patients with geriatric depression. AB - OBJECTIVE: The authors assessed frontotemporal function in patients with geriatric depression, a debilitating and increasingly prevalent disorder that has not been examined with brain activation paradigms. METHOD: Six depressed elderly patients and five healthy comparison subjects underwent high-sensitivity [(15)O]H(2)O positron emission tomography scans during a paced word generation task and a resting condition. RESULTS: Bilateral activation deficits were noted in the dorsal anterior cingulate gyrus and hippocampus of the depressed geriatric patients relative to the comparison subjects. Patients had memory deficits that correlated with lower hippocampal activity during both rest and activation. CONCLUSIONS: These initial findings suggest that hippocampal and dorsal anterior cingulate hypoactivation may constitute contributing neural substrates of geriatric depression. They also suggest that hippocampal dysfunction is related to the memory dysfunction characteristic of this disorder. PMID- 11481172 TI - Does the manic/mixed episode distinction in bipolar disorder patients run true over time? AB - OBJECTIVE: The authors sought to determine whether the manic/mixed episode distinction in patients with bipolar disorder runs true over time. METHOD: Over an 11-year period, the observed distribution of manic and mixed episodes (N=1,224) for patients with three or more entries in the management information system of a community mental health center (N=241) was compared to the expected distribution determined by averaging 1,000 randomly generated simulations. RESULTS: Episodes were consistent (all manic or all mixed) in significantly more patients than would be expected by chance. CONCLUSIONS: These data suggest a pattern of diagnostic stability over time for manic and mixed episodes in patients with bipolar disorder. Careful prospective studies of this issue are needed. PMID- 11481173 TI - Inverse relationship between serotonin 5-HT(1A) receptor binding and anxiety: a [(11)C]WAY-100635 PET investigation in healthy volunteers. AB - OBJECTIVE: The authors investigated the relationship between anxiety--a facet of the Revised NEO Personality Inventory dimension of neuroticism--and serotonin 5 HT(1A) receptor binding potential. METHOD: Positron emission tomography with [(11)C]WAY-100635 was used to estimate regional 5-HT(1A) binding potential in 19 healthy volunteers who completed the Revised NEO Personality Inventory. Correlation coefficients were calculated to determine the degree of association between 5-HT(1A) binding potential and personality inventory measures. RESULTS: There was a significant negative correlation between 5-HT(1A) binding potential and anxiety in four regions: the dorsolateral prefrontal cortex, anterior cingulate cortex, parietal cortex, and occipital cortex. CONCLUSIONS: The inverse relationship between 5-HT(1A) receptor binding potential and anxiety is consistent with 1) animal models that have shown higher anxiety in mice lacking 5 HT(1A) receptors and 2) clinical trial data that have demonstrated antianxiety properties of partial 5-HT(1A) agonists. PMID- 11481174 TI - Biperiden for excessive sweating from clozapine. PMID- 11481175 TI - Herbal diuretics and lithium toxicity. PMID- 11481176 TI - Zolpidem abuse. PMID- 11481177 TI - Flushing in a menopausal woman taking venlafaxine. PMID- 11481178 TI - Questions about reasons for living. PMID- 11481179 TI - Repeated self-mutilation and ECT. PMID- 11481180 TI - Classifying depression. PMID- 11481182 TI - Classifying depression. PMID- 11481183 TI - Late-onset schizophrenia-like psychosis. PMID- 11481185 TI - Aggression, serotonin, and seasonality. PMID- 11481187 TI - Risperidone, tardive dyskinesia, and the elderly. PMID- 11481189 TI - Lithium discontinuation: uncovering latent bipolar disorder? PMID- 11481193 TI - Genetic architecture of temperament. PMID- 11481195 TI - Yohimbine for anxiety disorders. PMID- 11481196 TI - Off-label uses of modafinil. PMID- 11481212 TI - Foundations for blockbuster drugs in federally sponsored research. AB - 'Blockbuster' drugs, which are widely prescribed and improve the health of millions, often originate in fundamental laboratory research. An important example of such drugs are the cholesterol-lowering drugs called 'statins', including Zocor, Pravachol, and Lipitor, which millions of people take in the U.S. every year. This short paper outlines the direct and indirect contributions of federally sponsored research to the development of these important drugs. PMID- 11481213 TI - Bone builders: preventing and treating osteoporosis. PMID- 11481214 TI - Bone builders: the discoveries behind preventing and treating osteoporosis. PMID- 11481215 TI - Cutaneous expression of corticotropin-releasing hormone (CRH), urocortin, and CRH receptors. AB - Studies in mammalian skin have shown expression of the genes for corticotropin releasing hormone (CRH) and the related urocortin peptide, with subsequent production of the respective peptides. Recent molecular and biochemical analyses have further revealed the presence of CRH receptors (CRH-Rs). These CRH-Rs are functional, responding to CRH and urocortin peptides (exogenous or produced locally) through activation of receptor(s)-mediated pathways to modify skin cell phenotype. Thus, when taken together with the previous findings of cutaneous expression of POMC and its receptors, these observations extend the range of regulatory elements of the hypothalamic-pituitary-adrenal axis expressed in mammalian skin. Overall, the cutaneous CRH/POMC expression is highly reactive to common stressors such as immune cytokines, ultraviolet radiation, cutaneous pathology, or even the physiological changes associated with the hair cycle phase. Therefore, similar to its central analog, the local expression and action of CRH/POMC elements appear to be highly organized and entrained, representing general mechanism of cutaneous response to stressful stimuli. In such a CRH/POMC system, the CRH-Rs may be a central element. PMID- 11481216 TI - A view through the clouds of imprinting. AB - The purpose of this review is to examine whether our current knowledge of the higher order control of gene expression and nuclear organization can help us understand the mechanisms of genomic imprinting. Imprinting involves the inheritance of a silenced allele of a gene through either a paternal or maternal germline. We have approached the problem of imprinting using a model based on the dynamic attachment of chromatin loops to immobilized RNA polymerases and control elements. We have combined the information from different experimental approaches, examining primarily the IGF2-H19 locus, in an attempt to simplify the complexity of the imprinting data that has accumulated. It is hoped that a unified model may generate predictions amenable to experimental testing and contribute to the interpretation of future experiments. PMID- 11481217 TI - Choline deficiency induces apoptosis in primary cultures of fetal neurons. AB - Treatment of rats with choline during brain development results in long-lasting enhancement of spatial memory whereas choline deficiency has the opposite effect. Changes in rates of apoptosis may be responsible. We previously demonstrated that choline deficiency induced apoptosis in PC12 cells and suggested that interruption of cell cycling due to a decrease in membrane phosphatidylcholine concentration was the critical mechanism. We now examine whether choline deprivation induces apoptosis in nondividing primary neuronal cultures of fetal rat cortex and hippocampus. Choline deficiency induced widespread apoptosis in primary neuronal cells, indicating that cells do not have to be dividing to be sensitive to choline deficiency. When switched to a choline-deficient medium, both types of cells became depleted of choline, phosphocholine and phosphatidylcholine, and in primary neurons neurite outgrowth was dramatically attenuated. Primary cells could be rescued from apoptosis by treatment with phosphocholine or lysophosphatidylcholine. As described previously for PC12 cells, an increase in ceramide (Cer) was associated with choline deficiency induced apoptosis in primary neurons. The primary neuronal culture appears to be an excellent model to explore the mechanism whereby maternal dietary choline intake modulates apoptosis in the fetal brain. PMID- 11481218 TI - Evidence for a second valve system in lymphatics: endothelial microvalves. AB - The mechanism for interstitial fluid uptake into the lymphatics remains speculative and unresolved. A system of intralymphatic valves exists that prevents reflow along the length of the lymphatic channels. However, these valves are not sufficient to provide unidirectional flow at the level of the initial lymphatics. We investigate here the hypothesis that initial lymphatics have a second, separate valve system that permits fluid to enter from the interstitium into the initial lymph channels but prevents escape back out into the tissue. The transport of fluorescent microspheres (0.31 microm) across endothelium of initial lymphatics in rat cremaster muscle was investigated with micropipette manipulation techniques. The results indicate that microspheres can readily pass from the interstitium across the endothelium into the lumen of the initial lymphatics. Once inside the lymphatic lumen, the microspheres cannot be forced out of the lumen even after elevation of the lymphatic pressure by outflow obstruction. Reaspiration of the microspheres inside the lymphatic lumen with a micropipette is blocked by the lymphatic endothelium. This blockade exists whether the aspiration is carried out at the microsphere entry site or anywhere along the initial lymphatics. Nevertheless, puncture of the initial lymphatic endothelium with the micropipette leads to rapid aspiration of intralymphatic microspheres. Investigation of lymphatic endothelial sections fixed during lymph pumping shows open interendothelial junctions not found in resting initial lymphatics. These results suggest that initial lymphatics have a (primary) valve system at the level of the endothelium. In conjunction with the classical (secondary) intralymphatic valves, the primary valves provide the mechanism that facilitates the unidirectional flow during periodic compression and expansion of initial lymphatics. PMID- 11481219 TI - Cardiac phosphodiesterase 5 (cGMP-specific) modulates beta-adrenergic signaling in vivo and is down-regulated in heart failure. AB - Recent studies implicate increased cGMP synthesis as a postreceptor contributor to reduced cardiac sympathetic responsiveness. Here we provide the first evidence that modulation of this interaction by cGMP-specific phosphodiesterase PDE5A is also diminished in failing hearts, providing a novel mechanism for blunted beta adrenergic signaling in this disorder. In normal conscious dogs chronically instrumented for left ventricular pressure-dimension analysis, PDE5A inhibition by EMD82639 had modest basal effects but markedly blunted dobutamine-enhanced systolic and diastolic function. In failing hearts (tachypacing model), however, EMD82639 had negligible effects on either basal or dobutamine-stimulated function. Whole myocardium from failing hearts had 50% lower PDE5A protein expression and 30% less total and EMD92639-inhibitable cGMP-PDE activity. Although corresponding myocyte protein and enzyme activity was similar among groups, the proportion of EMD82639-inhibitable activity was significantly lower in failure cells. Immunohistochemistry confirmed PDE5A expression in both the vasculature and myocytes of normal and failing hearts, but there was loss of z band localization in failing myocytes that suggested altered intracellular localization. Thus, PDE5A regulation of cGMP in the heart can potently modulate beta-adrenergic stimulation, and alterations in enzyme localization and reduced synthesis may blunt this pathway in cardiac failure, contributing to dampening of the beta-adrenergic response. PMID- 11481220 TI - Contribution of endogenously expressed Trp1 to a Ca2+-selective, store-operated Ca2+ entry pathway. AB - Heterologous expression of the transient receptor potential-1 gene product (Trp1) encodes for a Ca2+ entry pathway, though it is unclear whether endogenous Trp1 contributes to a selective store-operated Ca2+ entry current. We examined the role of Trp1 in regulating both store-operated Ca2+ entry and a store-operated Ca2+ entry current, I(SOC), in A549 and endothelial cells. Twenty different 'chimeric' 2'-O-(2-methoxy)ethylphosphothioate antisense oligonucleotides were transfected separately using cationic lipids and screened for their ability to inhibit Trp1 mRNA. Two hypersensitive regions were identified, one at the 5' end of the coding region and the second in the 3' untranslated region beginning six nucleotides downstream of the stop codon. Antisense oligonucleotides stably decreased Trp1 at concentrations ranging from 10 to 300 nM, for up to 72 h. Thapsigargin increased global cytosolic Ca2+ and activated a I(SOC), which was small (-35 pA @ -80 mV), reversed near +40 mV, inhibited by 50 microM La3+, and exhibited anomalous mole fraction dependence. Inhibition of Trp1 reduced the global cytosolic Ca(2+) response to thapsigargin by 25% and similarly reduced I(SOC) by 50%. These data collectively support a role for endogenously expressed Trp1 in regulating a Ca2+-selective current activated upon Ca2+ store depletion. PMID- 11481221 TI - Altered mitochondrial function and overgeneration of reactive oxygen species precede the induction of apoptosis by 1-O-octadecyl-2-methyl-rac-glycero-3 phosphocholine in p53-defective hepatocytes. AB - The mechanism of induction of apoptosis by the novel anti-cancer drug 1-O octadecyl-2-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3) was investigated in p53-defective SV40 immortalized rat hepatocytes (CWSV1). Exposure to 12 microM ET 18-OCH3 for 36 h induced apoptosis as determined using classical morphological features and agarose gel electrophoresis of genomic DNA. Increased levels of reactive oxygen species (ROS) were detected spectrophotometrically using a nitroblue tetrazolium (NBT) assay in cells treated with ET-18-OCH3. Both the increased generation of ROS and the induction of apoptosis were inhibited when cells were treated concurrently with ET-18-OCH3 in the presence of the antioxidant alpha-tocopherol. Similar results were achieved when cells were switched acutely to choline-deficient (CD) medium in the presence of the antioxidant. The possible role of mitochondria in the generation of ROS was investigated. Both ET-18-OCH3 and CD decreased the phosphatidylcholine (PC) content of mitochondrial and associated membranes, which correlated with depolarization of the mitochondrial membrane as analyzed using 5,5',6,6' tetramethylbenzimidazolcarbocyanine iodide (JC-1), a sensitive probe of mitochondrial membrane potential. Rotenone, an inhibitor of the mitochondrial electron transport chain, significantly reduced the intracellular level of ROS and prevented mitochondrial membrane depolarization, correlating with a reduction of apoptosis in response to either ET-18-OCH3 or CD. Taken together, these results suggest that the form of p53-independent apoptosis induced by ET-18-OCH3 is mediated by alterations in mitochondrial membrane PC, a loss of mitochondrial membrane potential, and the release of ROS, resulting in completion of apoptosis. PMID- 11481222 TI - Methamphetamine causes differential regulation of pro-death and anti-death Bcl-2 genes in the mouse neocortex. AB - Bcl-2, an inner mitochondrial membrane protein, inhibits apoptotic neuronal cell death. Expression of Bcl-2 inhibits cell death by decreasing the net cellular generation of reactive oxygen species. Studies by different investigators have provided unimpeachable evidence of a role for oxygen-based free radicals in methamphetamine (METH) -induced neurotoxicity. In addition, studies from our laboratory have shown that immortalized rat neuronal cells that overexpress Bcl-2 are protected against METH-induced apoptosis in vitro. Moreover, the amphetamines can cause differential changes in the expression of Bcl-X splice variants in primary cortical cell cultures. These observations suggested that METH might also cause perturbations of Bcl-2-related genes when administered to rodents. Thus, the present study was conducted to determine whether the use of METH might indeed be associated with transcriptional and translational changes in the expression of Bcl-2-related genes in the mouse brain. Here we report that a toxic regimen of METH did cause significant increases in the pro-death Bcl-2 family genes BAD, BAX, and BID. Concomitantly, there were significant decreases in the anti-death genes Bcl-2 and Bcl-XL. These results thus support the notion that injections of toxic doses of METH trigger the activation of the programmed death pathway in the mammalian brain. PMID- 11481223 TI - Inositol hexakisphosphate increases L-type Ca2+ channel activity by stimulation of adenylyl cyclase. AB - Inositol hexakisphosphate (InsP6) is a most abundant inositol polyphosphate that changes simultaneously with inositol 1,4,5-trisphosphate in depolarized neurons. However, the role of InsP6 in neuronal signaling is unknown. Mass assay reveals that the basal levels of InsP6 in several brain regions tested are similar. InsP6 mass is significantly elevated in activated brain neurons and lowered by inhibition of neuronal activity. Furthermore, the hippocampus is most sensitive to electrical challenge with regard to percentage accumulation of InsP6. In hippocampal neurons, InsP6 stimulates adenylyl cyclase (AC) without influencing cAMP phosphodiesterases, resulting in activation of protein kinase A (PKA) and thereby selective enhancement of voltage-gated L-type Ca2+ channel activity. This enhancement was abolished by preincubation with PKA and AC inhibitors. These data suggest that InsP6 increases L-type Ca2+ channel activity by facilitating phosphorylation of PKA phosphorylation sites. Thus, in hippocampal neurons, InsP6 serves as an important signal in modulation of voltage-gated L-type Ca2+ channel activity. PMID- 11481224 TI - Nitric oxide synthase expression and effects of nitric oxide modulation on contractility of rat extraocular muscle. AB - Extraocular muscles (EOMs) are specialized skeletal muscles that are constantly active, generate low levels of force for cross sectional area, have rapid contractile speeds, and are highly fatigue resistant. The neuronal isoform of nitric oxide synthase (nNOS) is concentrated at the sarcolemma of fast-twitch muscles fibers, and nitric oxide (NO) modulates contractility. This study evaluated nNOS expression in EOM and the effect of NO modulation on lateral rectus muscle's contractility. nNOS activity was highest in EOM compared with diaphragm, extensor digitorum longus, and soleus. Neuronal NOS was concentrated to the sarcolemma of orbital and global singly innervated fibers, but not evident in the multi-innervated fibers. The NG-nitro-L-arginine methyl ester (L-NAME, a NOS inhibitor), increased submaximal tetanic and peak twitch forces. The NO donors S-nitroso-N-acetylcysteine (SNAC) and spermineNONOate reduced submaximal tetanic and peak twitch forces. The effect of NO on the contractile force of lateral rectus muscle is greater than previously observed on other skeletal muscle. NO appears more important in modulating contraction of EOM compared with other skeletal muscles, which could be important for the EOM's specialized role in generation of eye movements. PMID- 11481225 TI - Glucose-6-phosphate dehydrogenase deficiency promotes endothelial oxidant stress and decreases endothelial nitric oxide bioavailability. PMID- 11481226 TI - Absence of adipocyte fatty acid binding protein prevents the development of accelerated atherosclerosis in hypercholesterolemic mice. PMID- 11481227 TI - An absolute requirement for Fyn in T cell receptor-induced caspase activation and apoptosis. PMID- 11481228 TI - Characterization of alpha-casozepine, a tryptic peptide from bovine alpha(s1) casein with benzodiazepine-like activity. PMID- 11481229 TI - cDNA microarray reveals altered cytoskeletal gene expression in space-flown leukemic T lymphocytes (Jurkat). PMID- 11481230 TI - Kainic acid induces selective mitochondrial oxidative phosphorylation enzyme dysfunction in cerebellar granule neurons: protective effects of melatonin and GSH ethyl ester. PMID- 11481231 TI - Interleukin 2 activates nuclear phospholipase Cbeta by mitogen-activated protein kinase-dependent phosphorylation in human natural killer cells. PMID- 11481232 TI - Glucocorticoid receptors are required for up-regulation of neuronal 5 lipoxygenase (5LOX) expression by dexamethasone. PMID- 11481233 TI - Resistance to HIV-1 infection among African sex workers is associated with global hyporesponsiveness in interleukin 4 production. PMID- 11481234 TI - Suppression of angiogenesis, tumor growth, and wound healing by resveratrol, a natural compound in red wine and grapes. PMID- 11481235 TI - Impairment of hypoxic pulmonary vasoconstriction in mice lacking the voltage gated potassium channel Kv1.5. PMID- 11481236 TI - Erythropoietin and erythropoietin receptors in the peripheral nervous system: changes after nerve injury. PMID- 11481237 TI - Regulation of myostatin by glucocorticoids after thermal injury. PMID- 11481238 TI - Convertases other than furin cleave beta-secretase to its mature form. PMID- 11481239 TI - Cytokine-activated human vascular smooth muscle delays apoptosis of neutrophils: relevance of interactions between cyclo-oxygenase-2 and colony-stimulating factors. PMID- 11481240 TI - Memory impairment and cholinergic dysfunction by centrally administered Abeta and carboxyl-terminal fragment of Alzheimer's APP in mice. PMID- 11481241 TI - Human beta-defensin 4: a novel inducible peptide with a specific salt-sensitive spectrum of antimicrobial activity. PMID- 11481242 TI - Aspirin inhibits NF-kappaB and protects from angiotensin II-induced organ damage. PMID- 11481243 TI - Expression and regulation of the mammalian SUMO-1 E1 enzyme. PMID- 11481244 TI - Nitric oxide mediates endotoxin-induced hypertriglyceridemia through its action on skeletal muscle lipoprotein lipase. PMID- 11481245 TI - Calcium sensors as new therapeutic targets for airway hyperresponsiveness and asthma. PMID- 11481246 TI - Pancreatic phospholipase A2 via its receptor regulates expression of key enzymes of phospholipid and sphingolipid metabolism. PMID- 11481247 TI - Regulation of the alpha-secretase ADAM10 by its prodomain and proprotein convertases. PMID- 11481248 TI - Disruption of the plasminogen activator inhibitor 1 gene reduces the adiposity and improves the metabolic profile of genetically obese and diabetic ob/ob mice. PMID- 11481249 TI - Integrin-linked kinase as a candidate downstream effector in proteinuria. PMID- 11481250 TI - Host nutritional selenium status as a driving force for influenza virus mutations. PMID- 11481251 TI - Selection of in vivo retrovirally transduced hepatocytes leads to efficient and predictable mouse liver repopulation. PMID- 11481252 TI - Reactive oxygen species enhance the migration of monocytes across the blood-brain barrier in vitro. PMID- 11481253 TI - Inhibition of cytokine production and interference in IL-2 receptor-mediated Jak Stat signaling by the hydroxylamine metabolite of sulfamethoxazole. PMID- 11481254 TI - Essential role for cholesterol in synaptic plasticity and neuronal degeneration. PMID- 11481255 TI - Growth hormone restores glucocorticoid-induced T cell suppression. PMID- 11481256 TI - Characterization of melanocortin-1 receptor gene variants in uveal melanoma patients. AB - PURPOSE: Allelic variations of the melanocortin-1 receptor (MC1R) gene have been linked to red hair and sun-sensitive skin types and may play a role in the susceptibility to develop cutaneous malignant melanoma (CMM). To define the role of MC1R gene in uveal melanoma, a case control study was performed, in which the presence of MC1R gene variations in uveal melanoma patients was compared with that of healthy controls. METHODS: MC1R gene variants were analyzed in 162 uveal melanoma patients and 255 healthy controls. After genomic DNA was isolated from venous blood, the MC1R gene was amplified by polymerase chain reaction (PCR) and examined for the presence of variants by single-strand conformation polymorphism (SSCP) analysis. Participants were asked to complete a questionnaire regarding skin type, eye color, and hair color. RESULTS: No disparity was found between the distribution of the MC1R gene variants in both groups. Furthermore, no associations between MC1R genotype and pigment phenotype were found. In contrast to CMM, uveal melanoma patients did not show specific MC1R gene variants. Compared with controls, most uveal melanoma patients had blue eyes (65%, P = 0.060) and skin type III (56%); however, in the uveal melanoma group the presence of dark blond hair was significantly elevated (46%, P = 0.030). These findings are in contrast with studies on CMM, where most patients have skin type II and red/fair hair. CONCLUSIONS: These data suggest that MC1R variants do not play a role in the susceptibility to develop uveal melanoma. Furthermore, most uveal melanoma patients share phenotypic characteristics that differ from findings in CMM patients. PMID- 11481257 TI - A role for the Tubby-like protein 1 in rhodopsin transport. AB - PURPOSE: To test the hypothesis that a lack of Tubby-like protein 1 (TULP1) function causes aberrant transport of nascent rhodopsin and to examine the functional relationship between the homologous proteins TULP1 and Tubby by studying mice carrying combined mutations. METHODS: Subcellular localization of TULP1 and rhodopsin in photoreceptors was determined by immunofluorescence and by postembedding immunoelectron microscopy. Mice carrying different tulp1/tubby allele combinations were examined by histology, electroretinograms (ERGs), and immunofluorescence microscopy. RESULTS: TULP1 is distributed throughout the photoreceptor cytoplasm but is excluded from the outer segments and the nuclei. In the tulp1-/- mice, ectopic accumulation of rhodopsin occurs at an early age. Both the vesicular profiles in the interphotoreceptor space and the inner segment plasma membranes are immunoreactive for rhodopsin. Mice doubly homozygous for null mutations in the tulp1 and tubby genes initially develop photoreceptors and express a battery of photoreceptor markers at age 14 days. Thereafter their photoreceptors undergo a fulminant degeneration that reaches completion by postnatal day 17. The disease phenotype in the double homozygote is much more severe than either single homozygote. Double heterozygotes are phenotypically normal. CONCLUSIONS: A lack of TULP1 function results in misrouting of nascent rhodopsin. TULP1 may be a component of the cellular machinery that targets nascent rhodopsin to the outer segments. Comparison of disease phenotypes in the single and double mutants suggests that TULP1 and Tubby are not functionally interchangeable in photoreceptors nor do they form an obligate functional complex. PMID- 11481258 TI - Active matrilysin (MMP-7) in human pterygia: potential role in angiogenesis. AB - PURPOSE: Pterygia are invasive, proliferative fibrovascular growths, with the matrix metalloproteinase (MMP) family of enzymes strongly implicated in the pathogenesis of these lesions. The purpose of this study was to determine the cellular distribution and activation status of matrilysin (MMP-7) in pterygia. METHODS: Resected pterygia (n = 8) and normal conjunctiva (n = 8) were sectioned and analyzed immunohistochemically with two different epitope-specific anti-MMP-7 monoclonal antibodies (Abs) which differentiate pro- and active MMP-7. The specificity of each Ab was confirmed by Western blot analysis of p aminophenylmercuric acetate (APMA)-activated and latent recombinant MMP-7. Pterygia (n = 4) and autologous normal conjunctiva (n = 4) were placed in organ culture to determine the activation status of secreted MMP-7. RESULTS: Precursor and active forms of MMP-7 were detected in epithelial cells from both pterygia and normal conjunctiva. Intense immunoreactivity for pro- and active MMP-7 was also observed in the pterygium vasculature, but was essentially absent from conjunctival vessels. Pro-MMP-7 was also identified in the epithelial basement membrane and associated with matrix components in pterygia. The 141-7B2 Ab reacted with the 30-kDa latent MMP-7, and the IM47L Ab precipitated a 19-kDa active enzyme, thus confirming the differential specificity of each Ab. Pro- and active MMP-7 were increased 1.4- and 2.7-fold, respectively, in the supernatants from organ-cultured pterygia compared with conjunctiva. CONCLUSIONS: This study is the first to specifically localize an active MMP species in pterygia and strengthens the hypothesis that these enzymes are involved in the pathogenesis of this disease. The data also suggest that MMP-7 may play a significant role in the angiogenesis that characterizes this lesion. Future studies will be directed at determining whether targeting MMP activity may be useful for treatment of pterygia. PMID- 11481259 TI - Improvement of HSV-1 necrotizing keratitis with amniotic membrane transplantation. AB - PURPOSE: Stromal herpes simplex virus keratitis (HSK) is an immune-mediated disease. Previous studies have indicated that T cells, neutrophils, and macrophages contribute to the tissue damage in HSK. It has been shown that human amniotic membrane promotes epithelial wound healing and has diverse anti inflammatory effects. In this study, the effect of amniotic membrane transplantation (AMT) on corneal wound healing and on inflammation in mice with necrotizing HSK was examined. METHODS: BALB/c mice were corneally infected with 10(5) plaque-forming units (PFU) of HSV-1 (KOS strain). In 16 mice that exhibited severe ulcerating HSK, the cornea was covered with a preserved human amniotic membrane as a patch. Corneas in 16 infected mice remained uncovered and served as a control. On days 2 and 7 after surgery, the amniotic membrane was removed (eight mice in each group), the HSV-1-infected cornea was evaluated clinically, and the eye was enucleated. Tissue sections were analyzed histologically for epithelialization and cellular infiltration and immunohistochemically with anti CD3 mAb to T cells, anti-CD11b mAb to both macrophages and neutrophils, or anti F4/80 mAb to macrophages. RESULTS: Profound regression of corneal inflammation and rapid closure of epithelial defects were observed clinically within 2 days in the amniotic membrane-covered eyes, whereas HSV-1 keratitis and ulceration progressed in all mice in the control group (P < 0.001). Histologically, corneal edema and inflammatory infiltration, and immunohistochemically the number of CD3(+), CD11b(+), and F4/80(+) cells in the cornea were markedly decreased at 2 and 7 days after amniotic membrane application, compared with the uncovered control corneas (P < 0.001). CONCLUSIONS: AMT promotes rapid epithelialization and reduces stromal inflammation and ulceration in HSV-1 keratitis. AMT in mice with HSV necrotizing stromal keratitis appears to be a useful model for investigating the effect and the action mechanism of human amniotic membrane. PMID- 11481260 TI - Rapid ocular angiogenic control via naked DNA delivery to cornea. AB - PURPOSE: To determine the efficacy and safety of naked plasmid gene therapy to the corneal stroma and epithelium. METHODS: Naked plasmid DNA was injected under pressure into the cornea of mice. The expression of genes coding for beta galactosidase (beta-gal), enhanced green fluorescent protein (EGFP), vascular endothelial growth factor (VEGF), and soluble Flt-1 (s-Flt) was recorded and measured with regard to dose, time course, and bioactivity. RESULTS: LacZ gene expression of the protein beta-gal was demonstrated as early as 1 hour, with expression persisting for 10 days. Plasmid-injected corneas remained clear and free of inflammation. EGFP was bicistronically expressed with VEGF to demonstrate the practicality of simultaneous in vivo analysis of gene expression and growth factor bioactivity. Corneal injection of a plasmid containing VEGF cDNA induced corneal and anterior chamber neovascularization. Moreover, corneal injection of plasmid containing the cDNA for the soluble form of the VEGF receptor Flt-1 effectively prevented corneal neovascularization. CONCLUSIONS: The cornea is readily accessible for gene therapy in the laboratory and in the clinic. The method described is safe, effective, titratable, and easily monitored. Naked DNA delivery to the cornea has the potential to alter the treatment of a wide variety of corneal and anterior segment diseases. PMID- 11481261 TI - Involvement of Sp1 elements in the promoter activity of genes affected in keratoconus. AB - PURPOSE: Keratoconus is a progressive disease that thins and scars the corneal stroma. In keratoconus corneas, levels of degradative enzymes, including lysosomal acid phosphatase (LAP) and cathepsin B, are elevated, and those of the inhibitors alpha1-proteinase inhibitor (alpha 1-PI) and alpha 2-macroglobulin (alpha 2-M) are reduced, especially in the epithelial layer. An increased expression of the transcription factor Sp1 was also demonstrated. The role of Sp1 in regulation of the genes affected in keratoconus was examined in this study. METHODS: DNA segments, containing 5'-flanking promoter sequences of the alpha 1 PI, LAP, cathepsin B, and alpha 2-M genes were ligated into the secreted alkaline phosphatase (SEAP) reporter gene vector. These constructs, along with the pSV beta-galactosidase control vector, were transfected into cultured human corneal epithelial and stromal cells and skin fibroblasts. Cotransfection with the Sp1 expression vector was performed in parallel. SEAP and beta-galactosidase enzyme activities were assayed. RESULTS: In corneal epithelial cells, as in stromal cells, alpha 1-PI promoter activity was suppressed by cotransfection of pPacSp1. The LAP, cathepsin B, and alpha 2-M promoters were functional in corneal cells, whereas activities of these promoters were much lower in skin fibroblasts. Cotransfection experiments indicated that the up- or downregulation of LAP, cathepsin B, and alpha 2-M observed in keratoconus-affected corneas was not mediated by Sp1. CONCLUSIONS: These results support the theory that the corneal epithelium, along with the stroma, is involved in keratoconus. An upstream role of Sp1 is indicated and the Sp1-mediated downregulation of the alpha 1-PI gene may be a key event in the disease development. PMID- 11481262 TI - Evidence for rectus extraocular muscle pulleys in rodents. AB - PURPOSE: Extraocular rectus muscle (EOM) pulleys are important determinants of orbital biomechanics in humans. In this study, the authors evaluated orbital connective tissue morphology, specifically characterizing rectus muscle pulleys, in the rat, a species with laterally placed eyes, afoveate vision, and a less complex visuomotor repertoire than primates. METHODS: Adult rat orbits were paraffin processed and serially sectioned for histochemical and immunohistochemical staining. Frozen sections of enucleated globes with intact EOMs and associated connective tissue were also studied with myosin immunohistochemistry and histochemistry for the mitochondrial enzyme, nicotinamide adenine dinucleotide (NADH)-tetrazolium reductase, to delineate the orbital layer relationship with the pulley tissue. RESULTS: Focal condensations of collagenous connective tissue were found in relationship to the rectus muscles in the equatorial Tenon's fascia, similar to those described as human recti muscle pulleys. The fibroelastic pulley rings were coupled to adjacent EOM pulleys by bands containing collagen and elastin. The coupling of pulleys to the orbital walls was significantly less than that previously described in humans. As in humans, there was a dual insertion of rodent rectus muscles, with the orbital layer inserting on the muscle pulley and the global layer attaching to the sclera. CONCLUSIONS: The data support the presence of structures in the rat orbit that are the morphologic equivalent of the human rectus pulley system. Although rodent and human pulleys were similar in many respects, there were species specific properties that may relate to established differences in orbital anatomy and/or visuomotor behavior. These data extend the rectus muscle pulley concept to rodents and may provide insight into pulley structure-function relationships. PMID- 11481263 TI - Detecting early glaucoma by assessment of retinal nerve fiber layer thickness and visual function. AB - PURPOSE: To compare the abilities of scanning laser polarimetry (SLP), optical coherence tomography (OCT), short-wavelength automated perimetry (SWAP), and frequency-doubling technology (FDT) perimetry to discriminate between healthy eyes and those with early glaucoma, classified based on standard automated perimetry (SAP) and optic disc appearance. To determine the agreement among instruments for classifying eyes as glaucomatous. METHODS: One eye of each of 94 subjects was included. Healthy eyes (n = 38) had both normal-appearing optic discs and normal SAP results. Glaucoma by SAP (n = 42) required a repeatable abnormal result (glaucoma hemifield test [GHT] or corrected pattern standard deviation [CPSD] outside normal limits). Glaucoma by disc appearance (n = 51) was based on masked stereoscopic photograph evaluation. Receiver operating characteristic (ROC) curve areas, sensitivities, and specificities were calculated for each instrument separately for each diagnosis. RESULTS: The largest area under the ROC curve was found for OCT inferior quadrant thickness (0.91 for diagnosis based on SAP, 0.89 for diagnosis based on disc appearance), followed by the FDT number of total deviation plot points of < or =5% (0.88 and 0.87, respectively), SLP linear discriminant function (0.79 and 0.81, respectively), and SWAP PSD (0.78 and 0.76, respectively). For diagnosis based on SAP, the ROC curve area was significantly larger for OCT than for SLP and SWAP. For diagnosis based on disc appearance, the ROC curve area was significantly larger for OCT than for SWAP. For both diagnostic criteria, at specificities of > or =90% and > or =70%, the most sensitive OCT parameter was more sensitive than the most sensitive SWAP and SLP parameters. For diagnosis based on SAP, the most sensitive FDT parameter was more sensitive than the most sensitive SLP parameter at specificities of > or =90% and > or =70% and was more sensitive than the most sensitive SWAP parameter at specificity of > or =70%. For diagnosis based on disc appearance at specificity of > or =90%, the most sensitive FDT parameter was more sensitive than the most sensitive SWAP and SLP parameters. At specificity > or = 90%, agreement among instruments for classifying eyes as glaucomatous was poor. CONCLUSIONS: In general, areas under the ROC curve were largest (although not always significantly so) for OCT parameters, followed by FDT, SLP, and SWAP, regardless of the definition of glaucoma used. The most sensitive OCT and FDT parameters tended to be more sensitive than the most sensitive SWAP and SLP parameters at the specificities investigated, regardless of diagnostic criteria. PMID- 11481264 TI - Identification of autoreactive T cells in Vogt-Koyanagi-Harada disease. AB - PURPOSE: To determine the finer specificity and immunologic features of autoreactive T cells in Vogt-Koyanagi-Harada (VKH) disease. METHODS: T-cell clones (TCCs ) specific to tyrosinase family proteins were raised from the peripheral blood mononuclear cells (PBMCs) of patients with VKH disease, and the response of the TCCs to 30-mer peptides was determined. The TCCs that were reactive to the peptides with strong binding sites for HLA DRB1*0405 were initially tested. Then, a finer specificity of these TCCs against 12- to 14-mer peptides was determined. The cytokine production of these clones was measured by ELISA. RESULTS: A total of 62 stable TCCs were established from the PBMCs of five patients with VKH (28 clones against tyrosinase, 34 clones against tyrosinase related protein [TRP]1). Five of 28 TCCs for tyrosinase and 2 of 34 for TRP1 were reactive to the 30-mer peptides with strong binding sites for HLA DRB1*0405. These seven clones showed proliferative responses to one or more of the 12- to 14 mer peptides that match the motif of the strong binding site for HLADRB1*0405. Five of seven of the TCCs may be T-helper (Th) type 1, one of the remaining TCCs may be Th0, and the other may be Th2. CONCLUSIONS: The autoreactive T cells against tyrosinase and/or TRP1 may contribute to the development of VKH disease. PMID- 11481265 TI - Rapid detection and typing of oculopathogenic strain of subgenus D adenoviruses by fiber-based PCR and restriction enzyme analysis. AB - PURPOSE: To develop a new detection and typing method of oculopathogenic strains of subgenus D adenoviruses directly from conjunctival scrapings by a combination of polymerase chain reaction (PCR) and restriction enzyme analysis (REA). METHODS: A new PCR method using primer pairs of AF2/AR2, which are specific for the fiber genes, were developed to amplify 1150-bp products from nine oculopathogenic prototypes of subgenus D adenoviruses. Amplicons were cleaved with three restriction enzymes: DdeI, HinfI, and RsaI. Clinical specimens of 102 conjunctival scrapings were also evaluated by this PCR method. Restriction patterns of prototypes were used for the typing of clinical samples. Detection limit was determined by the PCR amplification of a known amount of purified adenovirus serotype 8 DNA. RESULTS: A novel PCR method based on the fiber genes allowed the amplification of nine oculopathogenic serotypes of subgenus D (Ad8, Ad9, Ad15, Ad17, Ad19, Ad22, Ad28, Ad37, and Ad39). As little as 38.4 fg of adenovirus type 8 could be detected by this method. Positive results were obtained from 48 of 102 samples (47%) by both hexon- and fiber-based PCR, whereas only 29 of 102 (28.4%) yielded positive results by culture isolation/neutralization test (NT). All positive specimens (29 samples) of culture isolation and PCR-RFLP methods showed positive results by our new fiber based PCR method, and no positive products were detected from other subgenus of adenovirus or nonadenoviral DNA. CONCLUSIONS: A newly developed fiber-based PCR REA method for the detection and typing of adenoviruses is faster than any former PCR methods. This all-in-1-day detection and typing method will be quite useful to the rapid diagnosis of subgenus D adenovirus infection. PMID- 11481266 TI - Requirement of B7-mediated costimulation in the induction of experimental autoimmune anterior uveitis. AB - PURPOSE: To study the role of costimulatory signaling through the CD28-B7 interaction in experimental autoimmune anterior uveitis (EAAU). METHODS: Naive Lewis rats were immunized with insoluble melanin-associated antigen (MAA) derived from bovine iris and ciliary body. CTLA4-Fc, a recombinant protein comprised of the extracellular domain of human CTLA4 bound to mouse IgG2a Fc, was used to block the CD28-B7 interaction. A mutant version (CTLA4-Fc-mutant) was used as a control. The effect of CTLA4-Fc on the in vivo induction of disease with MAA was studied. Subsequently, the mechanism by which CTLA4-Fc blocked the interaction of CD28 and B7 was investigated in vivo, using the adoptive transfer of T cells derived from CTLA4-Fc-treated rats, and in vitro, using the proliferative response and cytokine production of MAA-T cells in the presence of CTLA4-Fc. RESULTS: CTLA4-Fc markedly reduced the incidence and severity of EAAU in Lewis rats after sensitization with MAA. The adoptive transfer of sensitized T cells from CTLA4-Fc-treated donors did not induce EAAU in naive recipients. CTLA4-Fc inhibited the expansion of antigen-specific MAA-T cells and the production of TNF alpha. CONCLUSIONS: The costimulatory signal delivered through CD28-B7 is required for the induction and pathogenesis of EAAU. In the absence of this signal, antigen-specific expansion of MAA reactive T cells as well as production of TNF-alpha is inhibited. Abrogation of this costimulatory signal may be an important therapeutic option for EAAU. PMID- 11481267 TI - The effects of intraocular injection of interleukin-13 on endotoxin-induced uveitis in rats. AB - PURPOSE: Interleukin (IL)-13 is a strong immunomodulatory cytokine that inhibits macrophages from secreting proinflammatory mediators. This study was conducted to investigate the effect of intraocular injection of IL-13 on the development of endotoxin-induced uveitis (EIU) in the Lewis rat. METHODS: One injection into the anterior chamber of recombinant human IL-13 (6 ng in 10 microl saline) was performed either simultaneously with a single injection of lipopolysaccharide (LPS) from Salmonella typhimurium into the footpad or 6 hours before the IL-13 injection. EIU was evaluated by slit lamp examination at 6, 16, and 24 hours after LPS injection. Counts of inflammatory cells were performed on cryostat sections after specific immunostaining. Anterior chamber paracentesis was performed, and kinetic analysis of the IL-13 injected in the anterior chamber was performed by ELISA. Cytokine and chemokine gene expression in the iris-ciliary body and the retina was evaluated by reverse transcription-polymerase chain reaction. RESULTS: A significant inhibition of ocular inflammation was observed in IL-13-treated rats at 16 and 24 hours after LPS injection. Unilateral injection of IL-13 inhibited EIU only in the injected eye. High levels of IL-13 were detected in the aqueous humor at 2 hours after local IL-13 injection to remain high up to 18 hours. In contrast, IL-13 was not detected in the corresponding sera. Quantitative analysis of inflammatory cells in ocular tissues showed a significant decrease in OX-42(+) cells (microglia, activated macrophages, dendritic cells, and polymorphonuclear leukocytes) and ED1(+) cells (monocytes-macrophages and dendritic cells) in treated rats. A decreased expression of TNF-alpha, IL-1 beta, IL-6, monocyte chemoattractant protein (MCP) 1, and macrophage inflammatory protein (MIP)-2 mRNAs was observed in the iris ciliary body and the retina from IL-13-treated rats, whereas IFN-gamma was upregulated in the iris-ciliary body. CONCLUSIONS: Injection of IL-13 into the anterior chamber may inhibit the ocular inflammation induced by LPS injection by reducing intraocular cytokine and chemokine mRNA expression in ocular tissues. PMID- 11481268 TI - Involvement of apoptosis and interferon-gamma in murine toxoplasmosis. AB - PURPOSE: A murine toxoplasmosis model has been developed that results in central nervous system (CNS) and ocular inflammation characterized by encephalitis with numerous brain tissue cysts and milder inflammation with rare tissue cysts in the eye after 4 weeks of Toxoplasma gondii infection. In this model IFN gamma and inducible nitric oxide (iNO) are protective against T. gondii infection. In this study, the role of apoptosis in the pathogenesis of toxoplasmosis was investigated. METHODS: C57BL/6 (wild-type mice), B6MRL/lpr, and B6MRL/gld (defective Fas or FasL expression, respectively) mice were infected intraperitoneally with 20 to 30 tissue cysts of the ME-49 strain of T. gondii. Mice were killed at days 0, 14, or 28 after infection. The eyes and brains were harvested for histologic, immunohistochemical, and molecular studies. Analysis included immunostaining for Fas, FasL, Bcl-2, and Bax; in situ apoptosis detection (TUNEL assay); RT-PCR amplification for IFN gamma; and measurement of ocular nitrite levels. The control mice were naive mice of each strain that received no inoculation or injection. RESULTS: Wild-type mice appeared to constitutively express apoptotic molecules at higher levels in the eye than in the brain. Consequently, during T. gondii infection, apoptosis was greater in the eyes than in the brain. Untreated naive lpr and gld mice showed no expression of Fas and FasL, respectively. After infection, a slightly higher number of tissue cysts (lpr, 11.8 +/- 2.4; gld, 10.3 +/- 3.4) were found in the brains of the mutants than in the control animals (8.8 +/- 2.9). However, no significant differences between the number of apoptotic cells, inflammatory scores, or number of tissue cysts were noted in the eyes. IFN gamma mRNA in control mice was detected at day 28 after infection, whereas in both mutants, mRNA production occurred earlier, at day 14. Ocular nitrite levels were higher in lpr and gld mice than in wild-type mice. CONCLUSIONS: No significant difference in the degree of ocular inflammation and apoptosis was detected between the wild-type and Fas or FasL mutant mice. However, there was an earlier and subjectively greater expression of IFN gamma in the brain and eye and a higher level of nitrite in the ocular tissue of mutant strains than in the wild type. Multiple factors are likely to be involved in the pathogenesis of ocular toxoplasmosis. PMID- 11481269 TI - Expression and putative role of 11 beta-hydroxysteroid dehydrogenase isozymes within the human eye. AB - PURPOSE: The human eye is an important target tissue for steroid hormones, and glucocorticoids have been implicated in the pathogenesis of ocular disease, including glaucoma. In peripheral tissues, corticosteroid hormone action is regulated at a prereceptor level through the activity of the 11 beta hydroxysteroid dehydrogenase (11 beta-HSD) isozymes: an oxo-reductase (11 beta HSD1) that activates cortisol (F) from cortisone (E) and a dehydrogenase (11 beta HSD2) that inactivates F to E. The purpose of this study was to analyze the expression and putative role of 11 beta-HSD within the human eye. METHODS: Immunohistochemical and reverse transcription-polymerase chain reaction (RT-PCR) studies were performed on sections of human ocular tissues, surgical trabecular meshwork (TM) specimens and a ciliary nonpigmented epithelial (NPE) cell-line. Free F and E concentrations in aqueous humor were determined by gas chromatography-mass spectrometry (GC/MS). IOP was measured in eight male volunteers before and after oral ingestion of carbenoxolone (CBX), a known inhibitor of 11 beta-HSD. RESULTS: 11 beta-HSD1 was expressed in the basal cells of the corneal epithelium and the NPE. 11 beta-HSD2 was restricted to the corneal endothelium. RT-PCR revealed mRNA for only the glucocorticoid receptor (GR) in the TM specimens, whereas GR, mineralocorticoid receptor and 11 beta-HSD1 mRNAs were all present in the NPE cell line. The demonstration of free F in excess of E (F/E 14:1) in the aqueous humor suggested predominant 11 beta-HSD1 activity. Compared with baseline (14.7 +/- 1.06 mm Hg, mean +/- SD), the IOP decreased significantly on both the third and seventh days of CBX ingestion (12.48 +/- 1.11 mm Hg, P < 0.0001 and 11.78 +/- 1.50 mm Hg, P < 0.0001, respectively). CONCLUSIONS: These results suggest that the 11 beta-HSD1 isozyme may modulate steroid-regulated sodium transport across the NPE, thereby influencing IOP. PMID- 11481270 TI - Scleral plug of biodegradable polymers containing ganciclovir for experimental cytomegalovirus retinitis. AB - PURPOSE: To evaluate the efficacy of a biodegradable scleral plug containing ganciclovir (GCV) in a rabbit model of human cytomegalovirus (HCMV) retinitis. METHODS: To develop a rabbit model for HCMV retinitis, HCMV solution was injected once into the vitreous cavity of pigmented rabbits. The treated animals were divided into three groups: group A received no treatment, group B was treated once with GCV solution, and group C was treated with a scleral plug containing GCV. Rabbits in group B received an intravitreal injection of GCV solution 1 week after HCMV inoculation. In group C, the scleral plug containing GCV was implanted in the vitreous of the rabbits 1 week after HCMV inoculation. Ophthalmoscopically, vitreoretinal findings in each group were graded from 0+ to 4+ every week for 4 weeks after HCMV injection. RESULTS: Eyes of group A rabbits showed whitish retinal exudates and vitreous opacities 3 days after HCMV inoculation. These materials increased gradually until 3 weeks after HCMV inoculation. Scores for vitreoretinal lesions were significantly lower in eyes of group B rabbits compared with those of group A at 1 week after GCV injection (P < 0.05). However, vitreoretinal inflammation in eyes of group B rabbits increased again thereafter, and no significant difference in inflammation between groups A and B was found 2 weeks after GCV injection. In eyes of group C, scores for vitreoretinal lesions were significantly lower compared with those in both group A and group B at 3 weeks after HCMV inoculation (P < 0.01). CONCLUSIONS: The results demonstrated that sustained release of GCV into the vitreous cavity with biodegradable scleral plugs was effective for the treatment of experimentally induced HCMV retinitis in rabbits. PMID- 11481271 TI - Potent alpha(2A)-adrenoceptor-mediated vasoconstriction by brimonidine in porcine ciliary arteries. AB - PURPOSE: An investigation into whether alpha(2)-adrenoceptor agonists induce contractions in the porcine ciliary arteries and to characterize the functional receptor subtype mediating these responses. METHODS: Isolated arteries from the intraocular part of the porcine ciliary artery were suspended in microvascular myographs for isometric tension recording. The segments were contracted with the alpha(2)-adrenoceptor agonists brimonidine, apraclonidine, and oxymetazoline. To determine which subtypes of the alpha(2)-adrenoceptor mediate this contraction, antagonists subselective for the different alpha(2)-adrenoceptors were added to the vessel bath before concentration-response curves for brimonidine were obtained. The following alpha(2)-adrenoceptor antagonists were applied: BRL44408 (alpha(2A)-selective), ARC239 (alpha(2B)- and alpha(2C)-selective), and prazosin (alpha(2B)- and alpha(2C)-selective). RESULTS: The alpha(2)-adrenoceptor agonists induced vasoconstriction in the porcine ciliary artery with the following potency order (EC(50)) expressed in nanomolar: brimonidine 2.11, oxymetazoline 5.26, and apraclonidine 13.0. As a reference, noradrenaline was tested, and its EC(50) was determined to be 247 nM in the ciliary artery. In the porcine ciliary arteries BRL44408, ARC239, and prazosin caused concentration-dependent and parallel rightward shifts of the concentration-response curves for brimonidine. Schild analyses for the antagonists against brimonidine yielded regression lines with slopes of unity and functional antagonist potencies (pK(B)) for BRL44408 (7.8), ARC 239 (5.8) and for prazosin (6.0) suggesting the presence of functional alpha(2A)-adrenoceptors. Moreover, there was a good correlation of pK(B) with ligand-binding affinity (pK(i)) of the alpha(2A)-adrenoceptor in the porcine eye tissue. CONCLUSIONS: The alpha(2)-adrenoceptor agonists brimonidine, apraclonidine, and oxymetazoline are potent vasoconstrictors in the porcine ciliary artery. In the present work, it was shown for the first time that the alpha(2A)-adrenoceptor subtype mediates this contraction. PMID- 11481272 TI - Nitric oxide attenuates alpha(2)-adrenergic receptors by ADP-ribosylation of G(i)alpha in ciliary epithelium. AB - PURPOSE: To determine the mechanism by which nitric oxide (NO) regulates alpha(2) adrenergic receptor coupling to adenylyl cyclase in bovine ciliary epithelium. METHODS: Ciliary epithelial explants were dissected, cultured, and labeled with [(3)H]adenine. [(3)H]Adenosine 3',5'-cyclic monophosphate (cAMP) was measured under basal conditions and after exposure to forskolin, isoproterenol, clonidine, yohimbine, pertussis toxin, and the NO donor spermine-NO. Endogenous and NO stimulated ADP-ribosylation of ciliary epithelial membrane proteins was determined by [(32)P]nicotinamide adenosine diphosphate (NAD) labeling and autoradiography. The three isoforms of the G(i)alpha protein subunit were evaluated by Western blot analysis. RESULTS: Basal [(3)H]cAMP content was 13.4 +/ 1.3 picomoles/mg protein (SEM). Both isoproterenol and forskolin stimulated [(3)H]cAMP accumulation to 36.0 +/- 3.9 and 73.2 +/- 17.5 picomoles/mg protein, respectively. Clonidine did not affect basal [(3)H]cAMP levels, but attenuated both isoproterenol- and forskolin-mediated [(3)H]cAMP accumulation to 23.2 +/- 4.4 and 31.6 +/- 4.6 picomoles/mg protein, respectively. Yohimbine antagonized the clonidine-mediated adenylyl cyclase inhibition. Pertussis toxin blocked the effect of clonidine. In the presence of the NO donor spermine-NO, the clonidine mediated inhibition of forskolin- and isoproterenol-stimulated cAMP accumulation was attenuated completely. NO significantly stimulated endogenous [(32)P]ADP ribosylation of a 40-kDa membrane protein. Western blot analysis with specific antibodies revealed expression of all three G(i) subtypes--G(i1)alpha, G(i2)alpha, and G(i3)alpha--in bovine ciliary epithelium. CONCLUSIONS: NO attenuates alpha(2)-adrenergic receptor-mediated inhibition of adenylyl cyclase in ciliary epithelium through ADP-ribosylation of the G(i)alpha subunit. The findings demonstrate heterologous regulation between the NO and cAMP signaling pathways in ciliary epithelium. PMID- 11481273 TI - Sensations evoked by selective mechanical, chemical, and thermal stimulation of the conjunctiva and cornea. AB - PURPOSE: To study the sensations evoked by selective mechanical, chemical, and thermal stimulation of the conjunctiva and compare them with those elicited by similar stimulation of the cornea. METHODS: Six young subjects participated in the study. Using a gas esthesiometer, selective mechanical (air puffs at flows from 0 to 264 ml/min), chemical (0--80% CO(2) in air), and thermal (air at temperatures from -10 degrees C to +80 degrees C) stimulation was performed on the center of the cornea and on the temporal conjunctiva. The intensity, degree of irritation, stinging and burning pain components, and thermal characteristics of the evoked sensation were evaluated after each stimulus in separate, 10-cm continuous visual analogue scales (VASs). The ability of the subjects to identify the quality of the stimulus applied to the cornea and the conjunctiva was also studied. RESULTS: The subjective intensity and thermal components (cooling or warming) of the sensation reported after mechanical, chemical, and heat stimulation were similar in the conjunctiva and cornea, although lower VAS scores were always reported in the conjunctiva for the irritation and the stinging and burning pain components. In the cornea, stimulation with low temperatures was perceived as a cooling sensation with an irritative component. In the conjunctiva, cooling was perceived as a purely cold sensation. Subjects showed similar discrimination capability in the cornea and the conjunctiva for the various types of stimuli. CONCLUSIONS: Sensations evoked in the cornea by selective mechanical, chemical, and heat and cold stimulation always presented an irritation component. In the conjunctiva, stimuli of the same intensity are always perceived as less irritating than in the cornea. Cold and other non noxious subqualities of sensation can be evoked in the conjunctiva. PMID- 11481274 TI - Proliferation, migration, and ERK activation in human retinal endothelial cells through A(2B) adenosine receptor stimulation. AB - PURPOSE: The nucleoside adenosine has been implicated in angiogenesis. A previous study demonstrated that activation of the A(2B) adenosine receptor (AdoR) increases cAMP accumulation, cell proliferation, and VEGF expression in human retinal endothelial cells (HRECs). In the present study, the role of this receptor was further characterized by examination of the effects of the selective A(2B) AdoR antagonists 3-N-propylxanthine (enprofylline) and 3-isobutyl-8 pyrrolidinoxanthine (IPDX) on AdoR-mediated HREC proliferation, capillary tube formation, and signal-transduction pathways. METHODS: HRECs were exposed to the adenosine analogue 5'-N-ethylcarboxamido-adenosine (NECA) in the absence or presence of AdoR antagonists. Migration was measured using Boyden chambers. Proliferation was assessed by counting cells. Western analysis was used to assess extracellular signal-related kinase (ERK) and cAMP response element-binding protein (CREB) in cell lysates. The effect of AdoR activation on tube formation was studied using cells grown on a synthetic basement membrane matrix. RESULTS: NECA induced proliferation in a concentration-dependent manner that was inhibited by enprofylline and IPDX. NECA stimulated chemotaxis in a concentration-dependent manner that was also blocked by both A(2B) AdoR antagonists. NECA activated ERK and CREB in HRECs. Both A(2B) AdoR antagonists diminished activation of ERK by NECA exposure. ERK activation was also blocked by the ERK-mitogen-activated protein kinase (MAPK) inhibitor PD98059, but not by the protein kinase A (PKA) inhibitor H-89. CREB activation was blocked by H-89, but not by PD98059, suggesting that ERK activation is independent of PKA. NECA enhanced tube formation on the matrix, whereas both A(2B) AdoR antagonists attenuated this effect. CONCLUSIONS: The selective A(2B) AdoR antagonists, enprofylline and IPDX, inhibited NECA-stimulated proliferation, ERK activation, cell migration, and capillary tube formation. A(2B) AdoR inhibition may offer a way to inhibit retinal angiogenesis and provide a novel therapeutic approach to treatment of diseases associated with aberrant neovascularization, such as diabetic retinopathy and retinopathy of prematurity. PMID- 11481275 TI - Neuroprotective effects of alpha(2)-selective adrenergic agonists against ischemia-induced retinal ganglion cell death. AB - PURPOSE: To investigate in adult rats the effects of two alpha(2)-selective adrenergic agonists (alpha(2)-SAs; AGN 191103 and AGN 190342) on retinal ganglion cell (RGC) survival after transient retinal ischemia. METHODS: RGCs were labeled with a Fluorogold (FG) tracer applied to both superior colliculi. Seven days later, the left ophthalmic vessels were ligated for 60 or 90 minutes. In one group, a single dose of saline or one alpha(2)-SA was administered intraperitoneally (IP) or topically 1 hour before ischemia. In another group, a single dose of AGN 190342 was administered IP, 1, 2, 4, 24, or 72 hours after ischemia. Rats were processed 7, 14, or 21 days later. Densities of surviving RGCs were estimated by counting FG-labeled cells in 12 standard retinal areas. RESULTS: Seven days after 60 or 90 minutes of retinal ischemia, death had occurred in 36% or 47%, respectively, of the RGC population, and by 21 days the loss of RGCs amounted to 42% or 62%, respectively. Systemic pretreatment with an alpha(2)-SA resulted in enhanced survival of ischemic-injured RGCs. Topical pretreatment with an alpha(2)-SA prevented up to 100% of the ischemia-induced RGC loss. Pretreatment with an alpha(2)-SA abolished the secondary slow RGC loss that occurred between days 7 and 21 after ischemia. When administered shortly after ischemia (up to 2 hours) AGN 190342 rescued substantial proportions of RGCs destined to die and diminished slow RGC death. CONCLUSIONS: Pretreatment and early posttreatment with an alpha(2)-SA induces marked long-lasting neuroprotective in vivo protection against ischemia-induced cell death in RGCs. PMID- 11481276 TI - Exudative retinal detachment from malignant uveal melanoma: predictors and prognostic significance. AB - PURPOSE: To determine independent predictors of exudative retinal detachment (RD) in eyes with uveal melanoma and the significance of RD in melanoma-specific survival. METHODS: The extent of exudative RD was recorded retrospectively in a population-based cohort of 167 consecutive patients with eyes enucleated from 1972 through 1981 because of choroidal and ciliary body melanoma, representing all melanomas treated during that period. Histopathologic features including microvascular loops and networks, microvascular density (MVD), and tumor infiltrating macrophages were determined. Clinical and histologic predictors of RD were modeled by multiple logistic regression with a split-sample, cross validation design. Survival was assessed by Kaplan-Meier analysis and adjusted for the effect of competing predictors by Cox proportional hazards regression. RESULTS: Of 142 (85%) eyes with adequate data, 25% had no RD, 16% had subretinal fluid around the tumor, 43% had clinical RD in one to two quadrants, and 16% had RD in three to four quadrants. The RD was more extensive if the tumor was large (P < 0.0001) and had microvascular loops and networks (P = 0.0094) and less extensive if it involved ciliary body (P = 0.011). High MVD (P = 0.054) and ruptured Bruch's membrane (P = 0.065) tended to be associated with RD. Multiple logistic regression showed largest basal diameter (odds ratio [OR] 1.43 for each 1-mm change, P < 0.0001), microvascular loops and networks (OR 1.95 for each category change, P = 0.0095), and ciliary body involvement (OR 0.20, P = 0.0039) to be independently associated with RD; ruptured Bruch's membrane (P = 0.96) and MVD (P = 0.87) were not associated. Clinical RD predicted poor survival (0.59 vs. 0.37 at 20 years; P = 0.029) by Kaplan-Meier analysis, but not after adjusting for other prognostic factors by Cox regression (hazard ratio [HR] 1.00, P = 1.0). CONCLUSIONS: Tumor size, which may be a surrogate measure for total vascular content and decompensation of choriocapillaris and retinal pigment epithelium, is a strong predictor of exudative RD. Microvascular loops and networks are likewise associated with exudative RD. Exudative RD is not associated with survival after adjusting for tumor size and microvascular loops and networks. PMID- 11481277 TI - High-frequency attenuation of the cone ERG and ON-response deficits in X-linked retinoschisis. AB - PURPOSE: Individuals with X-linked retinoschisis (XLRS) show a comparatively greater reduction of the ON response than the OFF response of the electroretinogram (ERG) of the cone system. At high temporal frequencies, they also show a marked attenuation of the flicker ERG that has been attributed to an abnormal cone photoreceptor response. The purpose of this study was to determine whether the high-frequency response attenuation contributes to the abnormal ERG ON response in XLRS. METHODS: Light-adapted ERGs were recorded from three patients with XLRS and from three control subjects, by using rapid-on and rapid off sawtooth flicker to emphasize ON and OFF responses, respectively, and by using low-pass sawtooth flicker, from which the high temporal frequencies had been removed to mimic the high-frequency attenuation in XLRS. RESULTS: For the control subjects, removing the high stimulus frequencies reduced the amplitude of the b-wave component of the ON response but had little effect on the amplitude of the d-wave component of the OFF response. In the patients with XLRS, the b-wave component of the ON response was already diminished using the full sawtooth stimulus, and removing the higher stimulus frequencies had no further effect. Patients' ERG responses to the 16-Hz stimulus fundamental alone were also abnormal, in that an initial response component normally present in the ERG was absent. CONCLUSIONS: The overall pattern of findings indicates that two factors contribute to the preferential ON-response deficit in XLRS: first, a high frequency attenuation of the cone photoreceptor response that effectively produces a low-pass stimulus for the postreceptoral pathway and that affects the ON response more than the OFF response and, second, a relatively greater attenuation of the ON- than of the OFF-bipolar cell response that is evident in the aberrant response to the sawtooth fundamental. PMID- 11481278 TI - In vivo evaluation of platelet--endothelial interactions after transient retinal ischemia. AB - PURPOSE: Accumulating evidence suggests that platelets play an important role in ischemia-reperfusion injury. To fulfill that role, platelets flowing in the bloodstream would have to interact with retinal endothelial cells and to accumulate in the postischemic retina. This study was designed to investigate quantitatively platelet-endothelial interactions in postischemic retina after transient retinal ischemia. METHODS: Transient retinal ischemia was induced in Long-Evans rats for 60 minutes by temporal ligation of the optic nerve. Isolated platelet samples labeled with carboxyfluorescein diacetate succinimidyl ester were administered intravenously to recipient rats after various reperfusion periods. Platelet-endothelial interactions in postischemic retina were evaluated in vivo with a scanning laser ophthalmoscope. Anti-P-selectin monoclonal antibody (mAb) was administered 5 minutes before the injection of labeled platelets. P selectin gene expression in the postischemic retina was studied by semiquantitative polymerase chain reaction. RESULTS: Under basal conditions, infused platelets showed minimal interactions with retinal endothelial cells. In contrast, postischemic retinas showed active platelet-endothelial interactions. Many platelets were observed rolling along and adhering to the major retinal veins. The number of rolling and adhering platelets reached a peak (555 +/- 65/mm per min and 25.8 +/- 3.2/mm(2)) 12 hours after reperfusion. However, the interactions between platelets and postischemic retinal endothelial cells were substantially inhibited by neutralizing P-selectin expressed on endothelial cells. In addition, P-selectin gene expression in postischemic retina corresponded with the time course of platelet-endothelial interactions during the reperfusion period. CONCLUSIONS: This study demonstrated that platelets actively interacted with retinal endothelial cells in the postischemic retina through P selectin expressed on the retinal endothelial cells. PMID- 11481279 TI - Normalization of retinal vascular permeability in experimental diabetes with genistein. AB - PURPOSE: To study the effects of genistein, a tyrosine kinase inhibitor, on retinal vascular permeability in an experimental diabetic rat model. METHODS: Seventy-two rats were equally divided into four groups: (1) nondiabetic control group, (2) diabetic control group, (3) diabetic rats receiving 150 mg genistein/kg food, and (4) diabetic rats receiving 300 mg genistein/kg food. Diabetes was induced by streptozotocin injection in the three diabetic groups. Rats were fed diets with or without genistein and followed for 6 months. Retinal vascular permeability was assessed by measuring radiolabeled sucrose leakage into the retina and by Western blot analysis for total retinal albumin. Retinal phosphotyrosine levels and proliferating cell nuclear antigen (PCNA) were also evaluated by Western blot analysis. RESULTS: Diabetic control rats had markedly increased retinal vascular leakage of radiolabeled sucrose compared with nondiabetic control rats. Diabetic rats receiving oral genistein had significantly less retinal vascular leakage of radiolabeled sucrose than diabetic control rats in a dose-response fashion. Diabetic control rats had increased levels of phosphotyrosine, retinal albumin, and PCNA by Western blot analysis compared with nondiabetic control rats. Rats receiving 300 mg of genistein had decreased retinal albumin by Western blot analysis. Western blot analysis demonstrated a dose-response decrease in retinal phosphotyrosine levels and PCNA in genistein-treated diabetic rats compared with diabetic control rats. CONCLUSIONS: Long-term oral administration of genistein significantly inhibits retinal vascular leakage in experimentally induced diabetic rats. Tyrosine kinase inhibition may be a useful pharmacological approach for the treatment of diabetic induced retinal vascular leakage. PMID- 11481280 TI - Cones regenerate from retinal stem cells sequestered in the inner nuclear layer of adult goldfish retina. AB - PURPOSE: To determine whether retinal progenitor cells in the inner nuclear layer give rise to regenerated cones after laser ablation of photoreceptors in adult goldfish retina. METHODS: Using a technique developed previously in this laboratory, photoreceptors in the retina of adult goldfish were ablated with an argon laser. The mitotic marker, bromodeoxyuridine, was used to label proliferating and regenerated cells, which were identified with cell-specific markers. RESULTS: Cells proliferating locally within lesion included microglia, Muller glia, and retinal progenitors in the inner nuclear layer (INL). The nuclei of both Muller glia and associated retinal progenitors migrated from the inner to the outer nuclear layer. The proliferating retinal progenitors, which express Notch-3 and N-cadherin, regenerated cone photoreceptors and then rod photoreceptors. CONCLUSIONS: Previous work has demonstrated that photoreceptors in the goldfish retina regenerate selectively after laser ablation, but the source of regenerated cones has not been identified. The results reported here provide support for the existence of retinal stem cells within the adult fish retina that are capable of regenerating cone photoreceptors. The data also support the involvement of Muller glia in the production of regenerated cones. PMID- 11481281 TI - Expression of the insulin receptor in the retina of the goldfish. AB - PURPOSE: Insulin is a peptide growth factor that is active in most tissues, both during development and in adulthood. The action of insulin is through its specific membrane receptor. Previously retinal progenitors in the adult goldfish were shown to proliferate vigorously when exposed to insulin in vitro.(1) The present study was undertaken to clone and characterize partial cDNAs that encode the goldfish's insulin receptor (IR) and to establish the cellular pattern of expression of this gene in the retina. METHODS: Standard methods were used for RNA isolation, reverse transcription-polymerase chain reaction, Northern blot analysis, and in situ hybridization. RESULTS: Multiple clones were isolated that, based on sequence analysis, segregated into two groups, presumed to represent two genes that encode the IR. These clones were designated goldfish IR-1 (gfIR-1) and goldfish IR-2 (gfIR-2). Northern blot analysis showed that both genes are expressed in multiple tissues, including the retina. Both gfIR-1 and -2 give rise to a single, major transcript, but the sizes of the two transcripts are different. In situ hybridizations using digoxygenin-labeled riboprobes showed that gfIR-1 and -2 are expressed by all differentiated retinal neurons as well as neuronal progenitors in the circumferential germinal zone. CONCLUSIONS: These data demonstrate that the IR is expressed in the retina of the goldfish, and, on the basis of the cellular pattern of expression, suggest that insulin may act both to regulate neurogenesis and influence the function of differentiated neurons. The cellular coexpression of the receptors for both insulin-like growth factor (IGF) 1 and insulin suggests that neurons and/or neuronal progenitors in the retina of the goldfish may contain hybrid IGF-1/insulin receptors. PMID- 11481282 TI - Regulated heat shock protein 27 expression in human retinal pigment epithelium. AB - PURPOSE: To examine the expression and regulation of an injury-related protein, heat shock protein (Hsp) 27, in retinal pigment epithelium (RPE), since RPE injury may be an important feature of age-related macular degeneration (ARMD). METHODS: Retinal cross sections from eyes of Lewis rats were examined for Hsp27 in vivo by immunohistochemistry, and in vitro expression of Hsp27 in human ARPE 19 cells was determined by Northern and Western blot analysis. Oxidant-mediated injury was performed by exposing ARPE-19 cells to myeloperoxidase and hydrogen peroxide. Cell lines stably expressing green fluorescent protein (GFP) targeted to the cell membrane were used to study injury-induced membrane blebbing, and XTT conversion was used to detect cell viability. RESULTS: High level of Hsp27 expression was detected in vivo in ganglion cells, RPE, and photoreceptor outer segments of rat retina. ARPE-19 cells also expressed high levels of Hsp27 in vitro. Oxidative injury in ARPE-19 cells resulted in transcriptional and translational activation of Hsp27 and induced extensive membrane blebbing. A high level of Hsp 27 protein was detected within membrane blebs. Increased expression of Hsp27 was also observed in differentiated ARPE-19 cells when compared with dividing cells. Higher Hsp27 levels in differentiated RPE cells correlated with increased viability and phenotypically different blebbing after exposure to the injury stimulus. In addition, sublethal injury doses caused a moderate amount of membrane blebbing, which was well tolerated by differentiated ARPE-19 cells. CONCLUSIONS: These results indicate that Hsp27 may be an important component of the RPE injury response and may contribute to injury-induced membrane blebbing in differentiated RPE cells. It is hypothesized that Hsp27 levels may play a role in disease states in the retina, such as ARMD. PMID- 11481283 TI - Oxidative stress--induced single-strand breaks in chromosomal telomeres of human retinal pigment epithelial cells in vitro. AB - PURPOSE: To demonstrate that chronic hyperoxia induces single-stranded breaks in chromosomal telomeres as a measure of oxidative DNA damage in cultured RPE cells. METHODS: RPE340 cells were cultured in 40% and 20% (control) O(2). DNA damage was assessed by mean terminal restriction fragment (TRF) length, and the S1 nuclease assay was used to determine the frequency of single-strand breaks in telomeric DNA. The degree of oxidative stress in cells was estimated by flow cytometric analysis of reactive oxygen intermediate (ROI)-induced 2',7' dichlorodihydrofluorescein diacetate fluorescence and Northern blot analysis of heme oxygenase-1 (HO-1) mRNA induction. RESULTS: The mean TRF length of cells grown in 40% O(2) shortened at a faster rate than those grown in 20% O(2). The S1 nuclease assay showed that the accelerated mean TRF length shortening was due to an increased accumulation of single-stranded breaks in telomeric DNA. The degree of ROI production and HO-1 mRNA induction was greater in cells treated with 40% than 20% O(2), an effect that was also larger in old than young passaged cells. CONCLUSIONS: RPE340 cells in vitro grown in chronic hyperoxia exhibited evidence of DNA damage with accelerated telomeric shortening via an increased accumulation of single-strand breaks in telomeric DNA. These changes could provide insight into aging of RPE cells by oxidative DNA damage. PMID- 11481284 TI - Electroencephalogram-based scaling of multifocal visual evoked potentials: effect on intersubject amplitude variability. AB - PURPOSE: The interindividual variability of the visual evoked potential (VEP) has been recognized as a problem for interpretation of clinical results. This study examines whether VEP variability can be reduced by scaling responses according to underlying electroencephalogram (EEG) activity. METHODS: A multifocal objective perimeter provided different random check patterns to each of 58 points extending out to 32 degrees nasally. A multichannel VEP was recorded (bipolar occipital cross electrodes, 7 min/eye). One hundred normal subjects (age 58.9 +/- 10.7 years) were tested. The amplitude and inter-eye asymmetry coefficient for each point of the field was calculated. VEP signals were then normalized according to underlying EEG activity recorded using Fourier transform to quantify EEG levels. High alpha-rhythm and electrocardiogram contamination were removed before scaling. RESULTS: High intersubject variability was present in the multifocal VEP, with amplitude in women on average 33% larger than in men. The variability for all left eyes was 42.2% +/- 3.9%, for right eyes 41.7% +/- 4.4% (coefficient of variability [CV]). There was a strong correlation between EEG activity and the amplitude of the VEP (left eye, r = 0.83; P < 0.001; right eye, r = 0.82; P < 0.001). When this was used to normalize VEP results, the CVs dropped to 24.6% +/- 3.1% (P < 0.0001) and 24.0% +/- 3.2% (P < 0.0001), respectively. The gender difference was effectively removed. CONCLUSIONS: Using underlying EEG amplitudes to normalize an individual's VEP substantially reduces intersubject variability, including differences between men and women. This renders the use of a normal database more reliable when applying the multifocal VEP in the clinical detection of visual field changes. PMID- 11481285 TI - Susceptibility testing of pathogenic fungi with itraconazole: a process analysis of test variables. AB - A 2(10-5) fractional factorial model was used to investigate the influence of 10 process variables in broth microdilution susceptibility tests with itraconazole against eight isolates of Candida species and six isolates of filamentous fungi in two growth media. An analysis of variance (ANOVA) indicated that glucose concentration and incubation time both significantly influenced control turbidity optical density (OD) values for most of the Candida spp. isolates, while incubation in >10% CO(2) versus ambient air, incubation temperature and inoculum size significantly influenced these OD values for about half of the yeast isolates. Control OD values for the mould isolates were most influenced by incubation time and temperature, and by occlusion of the wells with an adhesive sticker. Three statistical approaches, ANOVA, rank transformation and Mann Whitney U-test, were used to assess the influence of the variable combinations on MIC, determined with a 50% growth reduction end-point. Incubation temperature and time, glucose concentration and inoculum size were the variables that most often affected susceptibility results to the level of statistical significance; however, the supplier of RPMI 1640 medium, the use of adhesive stickers and the atmosphere of incubation significantly influenced the MIC for some isolates. The medium used to prepare the test inoculum, the solvent used to prepare the stock solution and the shape of the microdilution plate wells significantly affected outcome, but only sporadically. A principal component analysis of the data matrix confirmed this order of relative influence of the test variables on the MIC. Since each fungal isolate responded differently to combinations of process variables in the test, we conclude that any unified method for antifungal susceptibility determination represents a compromise, rather than an idealized system. PMID- 11481286 TI - Ferrocene-chloroquine analogues as antimalarial agents: in vitro activity of ferrochloroquine against 103 Gabonese isolates of Plasmodium falciparum. AB - The in vitro activities of ferrochloroquine, chloroquine, quinine, mefloquine, halofantrine, amodiaquine, primaquine, atovaquone and artesunate were evaluated against Plasmodium falciparum isolates from children with uncomplicated malaria from Libreville (Gabon), using an isotopic, micro, drug susceptibility test. The IC(50) values for ferrochloroquine were in the range 0.43-30.9 nM and the geometric mean IC(50) for the 103 isolates was 10.8 nM (95% CI 8.6-13.5 nM), while the geometric means for chloroquine, quinine, mefloquine, amodiaquine and primaquine were 370 nM, 341 nM, 8.3 nM, 18.1 nM and 7.6 microM, respectively. Ferrochloroquine was active against P. falciparum isolates, 95% of which showed in vitro resistance to chloroquine. Weak positive significant correlations were observed between the responses to ferrochloroquine and that to chloroquine, amodiaquine and quinine, but too low to suggest cross-resistance. There was no significant correlation between the response to ferrochloroquine and those to mefloquine, halofantrine, primaquine, atovaquone or artesunate. Ferrochloroquine may be an important alternative drug for the treatment of chloroquine-resistant malaria. PMID- 11481287 TI - Effects of trimethoprim and co-trimoxazole on the morphology of Listeria monocytogenes in culture medium and after phagocytosis. AB - The purpose of this study was to compare the extra- and intracellular activity of antifolates on Listeria monocytogenes. The fortuitous discovery of elongated bacteria in response to trimethoprim revealed a novel effect on the morphology of Listeria in cell culture medium and after phagocytosis. This phenomenon permitted the quantification of trimethoprim activity, revealing comparable activity intra- and extracellularly. Subinhibitory concentrations of trimethoprim resulted in bacterial elongation, which was reversed after removal of trimethoprim. We attribute this effect of trimethoprim to an inhibition of cell wall synthesis and/or cell separation of Listeria. PMID- 11481288 TI - The inhibition and selectivity of bacterial topoisomerases by BMS-284756 and its analogues. AB - Analogues of BMS-284756, a novel des-F(6)-quinolone, were synthesized and evaluated in order to determine the effects of modification of substituents on in vitro target inhibition. BMS-340281 (stereoisomer of BMS-284756), BMS-340280 (C-6 fluorinated analogue of BMS-284756), BMS-340278 (C-8-H derivative), BMS-433366 (C 8 methoxy analogue) and fluoroquinolone comparators were evaluated for antibacterial activity. The MICs of BMS-284756 were generally found to be within two-fold of the MICs of BMS-284756 analogues against a panel of Gram-positive and -negative organisms. BMS-284756 had MICs of 0.03-0.125 mg/L against Streptococcus pneumoniae strains with GyrA and ParC mutations, and was the most active quinolone. BMS-284756 and its analogues had similar activity compared with ciprofloxacin and moxifloxacin against topoisomerase IV decatenation, but were three times more active than levofloxacin. The IC(50) of BMS-284756 for human topoisomerase II (hTopo II) was 3000 times higher than its IC(50) for DNA gyrase, and no whole-cell cytotoxicity was noted. Two analogues, BMS-340280 and BMS 340278, demonstrated moderate inhibition against hTopo II and cytotoxicity in the cellular assay. BMS-284756 demonstrated greater Gram-positive antibacterial activity and similar inhibition of targets compared with other fluoroquinolones, and more favourable selectivity compared with the other BMS-284756 analogues. PMID- 11481289 TI - Short antibacterial peptides and erythromycin act synergically against Escherichia coli. AB - Five different peptides (6-18 residues) with chain lengths shorter than the required minimum to span the bacterial cell membrane as monomeric helices were designed in order to elucidate whether variation in chain length exerted differences in their mode of action. To gain a better understanding of the possible mode of action of these peptides, they were studied in combination with clinically used antibiotics with different targets. Antibiotic-peptide combinations were tested against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. No synergy was observed between the peptides and antibiotics when tested against S. aureus. Synergic interactions between all peptides and erythromycin were observed when tested against E. coli. Synergy was also observed with rifampicin and two peptides against E. coli. There was no clear-cut correlation between the ability to interact synergically or antagonistically and the number of residues. We further investigated the combined action of our peptides and PGLa, to elucidate peptide-peptide interactions. In contrast to previously reported synergy between magainin 2 and PGLa, our peptides did not show any synergy when combined with PGLa. Thus, our results indicate an alternative mode of action of these antibacterial peptides as compared with peptides such as magainin 2. PMID- 11481290 TI - In vitro activity of fosfomycin in combination with various antistaphylococcal substances. AB - Using the chequerboard technique we studied the in vitro activity of the broad spectrum antibiotic fosfomycin in combination with vancomycin, rifampicin, linezolid, quinupristin/ dalfopristin, cefazolin, meropenem and moxifloxacin against two Staphylococcus epidermidis strains (ATCC 12228, DSM 3269) and five Staphylococcus aureus isolates (ATCC 29213, DSM 683, DSM 46320, GISA 323/93, MRSA 3558/00). The phenomena of 'trailing' and 'skipped wells' did not present a problem. Synergy was the most common effect of all drugs tested in combination with fosfomycin; only combination with vancomycin showed antagonism for two of seven isolates. Using a killing-curve technique fosfomycin showed cidal activity, where increasing the drug concentration above the MIC did not enhance killing velocity. Inhibitory concentrations of vancomycin plus fosfomycin against DSM 46320 caused effects identical to those observed with vancomycin alone. The combination of fosfomycin plus linezolid exerted the bacteriostatic effect found with linezolid alone. Fosfomycin plus quinupristin/dalfopristin exhibited the bactericidal effect found with fosfomycin alone (in contrast to the rapidly bactericidal effect of quinupristin/dalfopristin). Electron microscopy showed that fosfomycin given in combination with linezolid, quinupristin/dalfopristin or moxifloxacin (substances that do not cause morphological alterations when given alone) resulted in 'cauliflower-shaped' distortion as caused by fosfomycin alone. Our in vitro data indicate considerable potential for fosfomycin used in combination with other antistaphylococcal antimicrobials, especially linezolid or quinupristin/dalfopristin. PMID- 11481291 TI - The in vitro activity of daptomycin against 514 Gram-positive aerobic clinical isolates. AB - The in vitro activity of daptomycin was assessed in comparison with that of vancomycin and penicillin against a wide range of Gram-positive aerobic clinical isolates. MICs were determined by an agar dilution method on Mueller-Hinton agar (NCCLS/EUCAST) and on Isotonic agar adjusted to contain 50 mg/L free calcium (BSAC). Both media were enriched with 5% horse blood for fastidious organisms. Daptomycin MICs for all 172 staphylococci, including methicillin-susceptible and methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus haemolyticus, were 0.03-0.5 mg/L. For 99 of the 100 enterococci (Enterococcus faecalis, n = 50; Enterococcus faecium, n = 50), including 37 vancomycin-resistant isolates, they were 0.25-2 mg/L. For all 108 beta-haemolytic streptococci, including Streptococcus pyogenes and Streptococcus agalactiae, daptomycin MICs were 0.016- 0.25 mg/L; for 101 alpha-haemolytic streptococci, including Streptococcus pneumoniae and 'viridans' streptococci, they were 0.016-2 mg/L. For miscellaneous vancomycin-resistant isolates including Lactobacillus spp., Lactococcus spp., Leuconostoc spp., Pediococcus spp. and isolates of Enterococcus casseliflavus and Enterococcus gallinarum, daptomycin MICs were 0.03 2 mg/L; MICs for the seven isolates of Listeria monocytogenes were 0.25-4 mg/L. There was little difference between the results on Mueller-Hinton agar and on supplemented Isotonic agar The discrepant results occasionally obtained tended to be one dilution higher on supplemented Isotonic agar. Daptomycin was active (MICs < or = 2 mg/L) against all the isolates tested with the exception of one isolate each of E. faecium and L. monocytogenes (MICs = 4 mg/L). Our results indicate that daptomycin MICs are independent of methicillin and vancomycin MICs. PMID- 11481292 TI - High proportion of pharyngeal carriers of commensal streptococci resistant to erythromycin in Spanish adults. AB - The presence of erythromycin-resistant (ErR) commensal streptococci in the throat of 110 healthy subjects and 87 patients with pharyngitis was investigated. The resistance determinants were studied by PCR using the primers for mef and erm genes, followed by hybridization and sequencing analysis. Overall, 94.4% of the subjects carried one or more ErR strains in their pharynx. A total of 253 ErR strains was studied: 127 (50.2%) showed constitutive or inducible resistance to clindamycin (MLS(B) phenotype) and 126 (49.8%) were susceptible to clindamycin (M phenotype). In 50 subjects (25.4%) both phenotypes were detected. The ermB gene was predominant among the MLS(B) phenotype strains (97.6%). The mefA (mefA/mefE) gene was detected in 100% of the strains with the M phenotype. One Streptococcus oralis strain bearing the MLS(B) phenotype carried both mefA and ermB genes. The mefA gene from clinical isolates of Streptococcus mitis and S. oralis was transferred by conjugation to an erythromycin-susceptible Streptococcus pneumoniae strain. PMID- 11481293 TI - Predominance of a methicillin-resistant Staphylococcus aureus clone susceptible to erythromycin and several other non-beta-lactam antibiotics in a Greek hospital. AB - A clone of heterogeneously methicillin-resistant Staphylococcus aureus (MRSA) isolates susceptible to erythromycin, ciprofloxacin, clindamycin, co-trimoxazole, nitrofurantoin, rifampicin, tetracycline and vancomycin, predominated in a Greek hospital with a high incidence of MRSA, representing 69.1% of the total MRSA isolates. All isolates of this clone lacked the conserved genes ermA and aadD. Two subtypes of this clone were detected, the more common being resistant to aminoglycosides and carrying the bi-functional gene aacA-aphD, while an aminoglycoside-susceptible variant, lacking this gene, lost a larger SmaI macrorestriction DNA fragment and gained a smaller fragment. PMID- 11481294 TI - Antimicrobial susceptibilities of Campylobacter strains isolated from food animals in Belgium. AB - Campylobacter spp. are a frequent cause of diarrhoea in man, originating mostly from poultry. It has been suggested that the veterinary use of antibiotics is largely responsible for resistance in human isolates, particularly to quinolones. During a 6 month period from June to December 1998, 677 Campylobacter isolates were obtained from healthy poultry and pigs. Samples were taken at Belgian slaughterhouses. Species identification was performed by biochemical tests, multiplex PCR and SDS-PAGE of whole-cell proteins. The in vitro susceptibility to six antimicrobial drugs was determined by the agar dilution method. Campylobacter jejuni was found more often in poultry than Campylobacter coli (79% C. jejuni versus 21% C. coli). In pigs the situation was reversed (6 versus 94%). Erythromycin resistance was significantly higher (P < 0.05) in C. coli, particularly in C. coli isolated from pigs (67.2%). Alarmingly high rates of resistance to ciprofloxacin were also noted, particularly for C. coli from broilers (62.1%). The latter indicates that resistance of Campylobacter in humans could derive from animals. PMID- 11481295 TI - Testing a degradable topical varnish of cetylpyridinium chloride in an experimental dental biofilm model. AB - Dental biofilms are highly associated with the development of dental caries. Novel drug delivery systems are being developed in order to eliminate cariogenic bacteria from the dental biofilms. We formulated two degradable sustained release varnishes, based on acrylic resin, with cetylpyridinium chloride (CPC) as the active agent. These formulations were tested in a dental biofilm model. The retention of CPC in the biofilm was dependent upon the pharmaceutical additives of the varnish. Both varnishes decreased bacterial adhesion, while also demonstrating marked antibacterial properties against the bacteria in the biofilm. PMID- 11481296 TI - Treatment of experimental candidosis with amphotericin B-Intralipid admixtures in immunocompromised mice. AB - The main goal of this research was the evaluation of the efficacy of amphotericin B (AMB) in comparison with AMB-Intralipid (AMB-IL) admixtures in cyclophosphamide (CY)-compromised animals for the treatment of systemic candidosis induced by several pathogenic Candida spp. Four-week-old ICR female mice were inoculated ip with 200 mg/kg of CY. At day 4 post-CY treatment the animals were inoculated iv with Candida albicans, Candida glabrata or Candida tropicalis (different inocula for the different species). Forty-eight hours later various doses of conventional AMB (0.4-1 mg/kg for 5 days) or AMB-IL admixtures (0.4-2 mg/kg for 5 days) were administered iv and the survival rate and mean survival time (MST) were evaluated during an observation period of up to 42 days. These experiments showed that while all control animals died, the survival rate of the AMB-treated mice ranged between 13 and 65% and that of the AMB-IL-treated mice was in the range 30-100% depending on the infecting dose and Candida species. The follow-up of the course of infection showed that AMB-IL admixtures increased the survival time of the treated mice. The MST was significantly higher for the mice treated with AMB-IL than for those treated with conventional AMB and was especially marked in the groups treated with high doses of the drug. Hence, the data obtained in the present study show that in CY-compromised mice AMB-IL admixtures were very effective in the treatment of systemic candidosis caused by C. albicans and non albicans species. PMID- 11481297 TI - Comparative evaluation of oral levofloxacin and parenteral nafcillin in the treatment of experimental methicillin-susceptible Staphylococcus aureus osteomyelitis in rabbits. AB - Methicillin-susceptible Staphylococcus aureus (MSSA) is the most common pathogen recovered from osteomyelitis patients. The current standard therapeutic method for acute phase osteomyelitis is parenteral antibiotic therapy. However, parenteral administration has negative aspects, such as secondary infection, patient inconvenience and high cost. The use of single oral antibiotic therapy may alleviate these problems. Therefore, the purpose of this study was to compare the effectiveness of standard once per day dosing of oral levofloxacin with a standard parenteral antibiotic regimen (nafcillin four times daily) for the treatment of experimental MSSA osteomyelitis in rabbits. Nearly all tibias from untreated infected controls (n = 27) revealed positive cultures (93%) for S. aureus, while the levofloxacin-treated group (n = 20) demonstrated significantly lower percentages of S. aureus infection (50%). The infected tibias of the nafcillin-treated group (n = 20) demonstrated significantly lower percentages (10%) of infected tibias than either the controls or the levofloxacin-treated groups (P < 0.05). The inferior efficacy of levofloxacin may have been due to the pharmacokinetic profile of this fluoroquinolone. The serum kinetics demonstrated that following single dose administration, levofloxacin was almost undetectable after 12 h. Studies in which levofloxacin is dosed every 12 h or given at increased doses in order to obtain bactericidal concentrations throughout the treatment regimen are needed. PMID- 11481298 TI - Pharmacokinetic and pharmacodynamic profile of high dose extended interval piperacillin-tazobactam. AB - A multiple-dose, open-labelled, randomized, two period crossover human volunteer study was performed (i) to describe the pharmacokinetic profile and safety profile of piperacillin and tazobactam (P/T) administered 6.0/0.75 g and 8.0/1.0 g q12h and (ii) to characterize the pharmacodynamic profile of these regimens against a variety of common targeted pathogens. Blood samples were collected after the third dose and concentrations of P/T were determined by a validated high-performance liquid chromatography assay. Pharmacokinetic profiles of P/T were determined by non-compartment analysis. Percentage time above the MIC (%T > MIC) of piperacillin was calculated for a range of MICs. In this study, no adverse events were attributed after multiple administrations of either 6.0/0.75 g or 8.0/1.0 g dose regimens. The peak concentration, half-life and area under the curve (AUC0-(0-tau)) of piperacillin were significantly different by a paired t-test (P < 0.05) between the two study regimens. The trough concentration, half life and area under the curve (AUC0-(0-tau)) of tazobactam were substantially different from parameters reported previously for conventional regimens. The 8.0/1.0 g regimen provided 50% T > MIC for MICs < or =32 mg/L, while a similar value for the 6.0/0.75 g regimen was < or = 16 mg/L. High-dose P/T regimens with extended interval were well tolerated and provide adequate dynamic exposure for a variety of susceptible pathogens. PMID- 11481299 TI - Effect of amoxycillin with or without clavulanate on adenoid bacterial flora. AB - The effect of antimicrobial therapy with amoxycillin (AMX) or co-amoxiclav (AMC) on the adenoid bacterial flora of 45 children with recurrent otitis media (ROM), scheduled for elective adenoidectomy, was studied. Patients were randomized before surgery into three groups of 15, having had either no antibiotic therapy (control), or 10 days of therapy with AMX or AMC. Core adenoid tissues were quantitatively cultured for aerobic and anaerobic bacteria. Polymicrobial aerobic anaerobic flora was present in all instances. The predominant aerobes in all groups were alpha-haemolytic and non-haemolytic streptococci, Haemophilus influenzae, Staphylococcus aureus, group A beta-haemolytic streptococci and Moraxella catarrhalis. The prominent anaerobes were Peptostreptococcus spp., Prevotella spp. and Fusobacterium spp. The number of isolates was significantly reduced in those treated with AMX (110; P < 0.05) or AMC (54; P < 0.001) compared with control (148). The number of bacteria per gram of tissue was lower in those treated with both antibiotics. The number of potential pathogens was lower in those treated with AMC compared with the other two groups (P < 0.001 ). The number of beta-lactamase- producing bacteria (BLPB) was lower in those treated with AMC compared with those treated with AMX (P < 0.025) or no antibiotic (P < 0.001). These data illustrate the ability of AMX and AMC to reduce the bacterial load as well as potential pathogens and BLPB from the adenoids of children with ROM. PMID- 11481300 TI - Erythromycin inhibits Pseudomonas aeruginosa-induced tumour necrosis factor-alpha production in human whole blood. AB - Erythromycin has been shown to be beneficial for panbronchiolitis, a disorder linked to infection with Pseudomonas aeruginosa. Erythromycin, but not the anti Pseudomonas antibiotics imipenem, ceftazidime, gentamicin and ciprofloxacin, caused a dose-dependent decrease in the production of tumour necrosis factor (TNF)-alpha by whole blood stimulated with heat-killed P. aeruginosa. The release of interleukin (IL)-10, IL-6, interferon-gamma and IL-8 was inhibited only at the highest erythromycin concentration. Inhibition of TNF-alpha production by erythromycin may, at least in part, explain the efficacy of this macrolide during panbronchiolitis despite its lack of activity for P. aeruginosa. PMID- 11481301 TI - In vitro activity of cefditoren against clinical isolates of penicillin susceptible and penicillin-intermediate strains of Streptococcus pneumoniae isolated in Germany, 1992-1998. AB - This study investigates the susceptibility to cefditoren of penicillin susceptible strains of invasive Streptococcus pneumoniae (n = 312) and of penicillin-intermediate strains of S. pneumoniae (n = 30) isolated mainly from patients with respiratory tract infections. The MIC(90)s of penicillin susceptible and -intermediate isolates were as follows: cefditoren, < or =0.06 and 1 mg/L; penicillin G, < or =0.06 and 0.5 mg/L. Cefditoren showed the highest activity against the penicillin-intermediate strains investigated compared with the other beta-lactam antibiotics and is therefore considered to be a promising agent for the treatment of infections caused by pneumococci with reduced penicillin susceptibility. PMID- 11481302 TI - In vitro activity of LY333328 (oritavancin) against Gram-positive aerobic cocci and synergy with ciprofloxacin against enterococci. AB - Gram-positive cocci are a major cause of nosocomial bacteraemias and are often resistant to most antibiotics. The emergence of enterococci with reduced susceptibility to vancomycin has created an urgent need for novel antibiotics to combat infections associated with these bacteria. In this study the in vitro activity of LY333328 (oritavancin), a semi-synthetic glycopeptide, was evaluated. LY333328 was effective against all strains of staphylococci, streptococci and enterococci tested. A combination of LY333328 and ciprofloxacin was additive for 50% of Enterococcus faecium strains resistant to both vancomycin and ciprofloxacin, and for 100% of vancomycin-susceptible and either ciprofloxacin susceptible or -resistant strains. PMID- 11481303 TI - A comparison of culture and PCR to determine the prevalence of ampicillin resistant bacteria in the faecal flora of general practice patients. AB - Of 95 faecal specimens containing bacterial DNA amplified by PCR, 24% contained cultivable bacteria that were resistant to high-level ampicillin. When these samples were examined by PCR using primers to amplify the bla(TEM) gene, the number of positive samples identified increased significantly to 49 (52%). These results indicate that ampicillin resistance is common in the study population. Furthermore, the bla(TEM) gene is more common than indicated by the recovery of resistant bacteria in culture. This points to potential anomalies in the assessment of the prevalence of resistance when relying on recovery of resistant bacteria by culture. PMID- 11481304 TI - Genetic and phenotypic characterization of macrolide resistance in group A streptococci isolated from adults with pharyngo-tonsillitis in France. AB - Three hundred and three strains of group A streptococci (GAS) isolated from adults with pharyngitis were tested to evaluate their phenotype of resistance to macrolides-lincosamides and to search for macrolide resistance genes. MICs of clarithromycin were determined. The overall rate of resistance to both erythromycin and clarithromycin was 9.6%. Constitutive, inducible and M phenotypes of resistance were detected in 4.3, 2 and 3.3% of strains, respectively. All constitutive phenotypes harboured ermB genes, whereas inducible phenotypes had the ermTR gene and M phenotypes had the mefA gene. In France, the current resistance rate of GAS to erythromycin and clarithromycin remains low. PMID- 11481305 TI - A 9 year study of clarithromycin and metronidazole resistance in Helicobacter pylori from Spanish children. AB - The evolution of clarithromycin, metronidazole and amoxycillin resistance in 246 Spanish Helicobacter pylori clinical isolates, obtained from paediatric patients during a 9 year period, was determined by an agar dilution technique. Clarithromycin resistance (MIC 1 mg/L) was 2.27% (IC95 0.05-12.02) in 1991-1993, 20.98% (IC95 12.72-31.46) in 1994-1996 and 28.33% (IC95 20.48-37.28) in 1997-1999 (P < 0.01). Metronidazole resistance (MIC 8 mg/L) was 7.14% (IC95 1.49-19.48) in 1991-1993, 20.25% (IC95 12.04-30.79) in 1994-1996 and 43.90% (IC95 32.95-55.30) in 1997-1999 (P < 0.01). Amoxycillin resistance was not found (all strains showed MICs < 2 mg/L). PMID- 11481306 TI - Dual nucleoside analogue treatment in the era of highly active antiretroviral therapy (HAART): a single-centre cross-sectional survey. AB - Since limited literature exists regarding the outcomes of dual nucleoside analogue reverse transcriptase inhibitor (NRTI) treatment in the highly active antiretroviral therapy (HAART) era, a cross-sectional survey was carried out in a population of around 1000 HIV-infected patients in a single northern Italian university hospital, to assess the frequency, background and long-term evolution of anti-HIV treatment conducted with two NRTIs. An appreciable proportion (20.4%) of the 798 HIV-infected patients currently treated with antiretrovirals at our centre still take a dual NRTI combination, and the great majority of them (68.7%) have a stable disease course (characterized by a viral load <3.7 log(10) HIV RNA copies/mL, a maintained CD4(+) lymphocyte count and absence of HIV disease progression after at least 24 months of follow-up), regardless of the selected regimen, prior antiretroviral therapy use, and baseline virological and immunological situation. Further studies are warranted to establish whether dual NRTI regimens may have residual indications in the HAART era, and whether the shift to a triple antiretroviral combination is expected to lead to long-term advantages in patients with a low risk of disease progression while on dual NRTI treatment. PMID- 11481307 TI - Use of fluconazole in daily practice: still room for improvement. AB - One hundred courses of fluconazole treatment in a university hospital and 81 courses in a non-university teaching hospital have been analysed in a prospective audit to evaluate prescribing practices. The quality of treatments was assessed by an infectious disease specialist and a pharmacist according to standard guidelines. In the non-university hospital, prescribed dosages were lower than in the university hospital, and often below the recommended dose. Mean duration of treatment for oesophageal candidosis and disseminated infections was considerably shorter in the non-university hospital compared with the university hospital, and often judged too short. Microbiological samples were examined in 75% of the cases in both hospitals. The expert reviewers agreed with the indication to use fluconazole in 58-100% of cases in the university hospital and 42-80% in the non university hospital, depending on the type of infection. There did not appear to be a major problem with inappropriate use of fluconazole. However, important issues for improvement could be identified, such as increasing the dosage and duration of treatment in cases of serious infections, and withholding treatment from patients with colonization rather than infection. PMID- 11481308 TI - The efficacy of continuous infusion flucloxacillin in home therapy for serious staphylococcal infections and cellulitis. AB - The efficacy and safety of continuous infusion flucloxacillin as home-based treatment was assessed in 62 consecutive patients with proven serious methicillin susceptible Staphylococcus aureus (MSSA) infections (n = 36) and cellulitis (n = 26). The treatment was well tolerated and resulted in cure or adequate suppression of infection in 27 of 28 (96%) patients in the serious MSSA infection group, and in 24 of 26 (92%) patients in the cellulitis group. PMID- 11481309 TI - Clindamycin treatment of Staphylococcus aureus expressing inducible clindamycin resistance. PMID- 11481310 TI - First clinical isolate of Streptococcus pneumoniae exhibiting high-level resistance to fluoroquinolones in Taiwan. PMID- 11481311 TI - Mechanisms of erythromycin and trimethoprim resistance in the Alaskan Streptococcus pneumoniae serotype 6B clone. PMID- 11481312 TI - Antibiotic resistance in Burkholderia cepacia at two regional cystic fibrosis centres in Northern Ireland: is there a need for synergy testing? PMID- 11481313 TI - Cefepime in vitro activity against derepressed extended-spectrum beta-lactamase (ESBL)-producing and non-ESBL-producing Enterobacter cloacae by a disc diffusion method. PMID- 11481314 TI - In vitro susceptibility testing of BMS-284756 by the BSAC standardized disc testing method. PMID- 11481315 TI - Ciprofloxacin susceptibility testing of enterococcal urinary isolates in accordance with BSAC guidelines. PMID- 11481316 TI - Serum pharmacokinetics and sputum penetration of amikacin 30 mg/kg once daily and of ceftazidime 200 mg/kg/day as a continuous infusion in cystic fibrosis patients. PMID- 11481317 TI - Gentamicin concentration in bile after once-daily versus thrice-daily dosing of 4 mg/kg/day. PMID- 11481318 TI - Three molecules of ubiquinone bind specifically to mitochondrial cytochrome bc1 complex. AB - Bifurcated electron flow to high potential "Rieske" iron-sulfur cluster and low potential heme b(L) is crucial for respiratory energy conservation by the cytochrome bc(1) complex. The chemistry of ubiquinol oxidation has to ensure the thermodynamically unfavorable electron transfer to heme b(L). To resolve a central controversy about the number of ubiquinol molecules involved in this reaction, we used high resolution magic-angle-spinning nuclear magnetic resonance experiments to show that two out of three n-decyl-ubiquinones bind at the ubiquinol oxidation center of the complex. This substantiates a proposed mechanism in which a charge transfer between a ubiquinol/ubiquinone pair explains the bifurcation of electron flow. PMID- 11481319 TI - Oxidative modification of tryptophan 43 in the heme vicinity of the F43W/H64L myoglobin mutant. AB - The F43W/H64L myoglobin mutant was previously constructed to investigate the effects of electron-rich tryptophan residue in the heme vicinity on the catalysis, where we found that Trp-43 in the mutant was oxidatively modified in the reaction with m-chloroperbenzoic acid (mCPBA). To identify the exact structure of the modified tryptophan in this study, the mCPBA-treated F43W/H64L mutant has been digested stepwise with Lys-C achromobacter and trypsin to isolate two oxidation products by preparative fast protein liquid chromatography. The close examinations of the (1)H NMR spectra of peptide fragments reveal that two forms of the modified tryptophan must have 2,6-disubstituted indole substructures. The (13)C NMR analysis suggests that one of the modified tryptophan bears a unique hydroxyl group in stead of the NH(2) group at the amino terminal. The results together with mass spectrometry (MS)/MS analysis (30 Da increase in mass of Trp-43) indicate that oxidation products of Trp-43 are 2,6 dihydro-2,6-dioxoindole and 2,6-dihydro-2-imino-6-oxoindole derivatives. Our finding is the first example of the oxidation of aromatic carbons by the myoglobin mutant system. PMID- 11481320 TI - Characterization of the rodent genes for arylacetamide deacetylase, a putative microsomal lipase, and evidence for transcriptional regulation. AB - In the current study, we have determined the cDNA and the genomic sequences of the arylacetamide deacetylase (AADA) gene in mice and rats. The AADA genes in the rat and mouse consist of five exons and have 2.4 kilobases of homologous promoter sequence upstream of the initiating ATG codon. AADA mRNA is expressed in hepatocytes, intestinal mucosal cells (probably enterocytes), the pancreas and also the adrenal gland. In mice, there is a diurnal rhythm in hepatic AADA mRNA concentration, with a maximum 10 h into the light (post-absorptive) phase. This diurnal regulation is attenuated in peroxisome proliferator-activated receptor alpha knockout mice. Intestinal but not hepatic AADA mRNA was increased following oral administration of the fibrate, Wy-14,643. The homology of AADA with hormone sensitive lipase and the tissue distribution of AADA are consistent with the view that AADA plays a role in promoting the mobilization of lipids from intracellular stores and in the liver for assembling VLDL. This hypothesis is supported by parallel changes in AADA gene expression in animals with insulin-deficient diabetes and following treatment with orotic acid. PMID- 11481321 TI - Induction of macrophage matrix metalloproteinase biosynthesis by surfactant protein D. AB - Recent studies strongly suggest that surfactant protein D (SP-D) plays important roles in pulmonary host defense and the regulation of immune and inflammatory reactions in the lung. Although SP-D can bind to alveolar macrophages and can elicit their chemotaxis, relatively little is known about the direct cellular consequences of SP-D on the function of these cells. Because matrix metalloproteinases (MMPs) are synthesized in increased amounts in response to various proinflammatory stimuli, we investigated the capacity of SP-D to modulate the production of MMPs by freshly isolated human alveolar macrophages. Unexpectedly we found that recombinant rat SP-D dodecamers selectively induce the biosynthesis of collagenase-1 (MMP-1), stromelysin (MMP-3), and macrophage elastase (MMP-12) without significantly increasing the production of tumor necrosis factor alpha and interleukin-1beta. SP-D did not alter the production of these MMPs by fibroblasts. Phosphatidylinositol, a surfactant-associated ligand that interacts with the carboxyl-terminal neck and carbohydrate recognition domains of SP-D, inhibited the SP-D-dependent increase in MMP biosynthesis. A trimeric, recombinant protein consisting of only the neck and carbohydrate recognition domain did not augment metalloproteinase production, suggesting that the stimulatory effect on MMP production depends on an appropriate spatial presentation of trimeric lectin domains. Although SP-D dodecamers can selectively augment metalloproteinase activity in vitro, this effect may be competitively inhibited by tissue inhibitors of metalloproteinases or surfactant-associated ligands in vivo. PMID- 11481322 TI - p300 Functions as a coactivator of transcription factor GATA-4. AB - Transcription factor GATA-4 plays critical roles in controlling heart development and cardiac hypertrophy. To understand how GATA-4 functions under diverse conditions, we sought to identify its coactivators. We tested p300 as a coactivator in GATA-4-dependent transient transcription assays in NIH3T3 cells and found that p300 synergistically activated GATA-4-dependent transcription on both synthetic and natural promoters. Direct physical interactions between the N- and C-zinc finger domains of GATA-4 and the cysteine/histidine-rich region 3 (C/H3) of p300 were identified in immunoprecipitation and glutathione S transferase pull-down experiments. Deletion of the C/H3 region of p300 abolished its coactivator activity indicating that the physical interaction was required for functional synergy. Through the use of a series of GATA-4 zinc finger mutants, the amino acids WRR in the C finger were identified as critical to the interaction. The adenoviral E1A protein or a peptide encoding the C/H3 region of p300 could inhibit GATA-4-dependent transcription, presumably by competing for p300 binding. Furthermore, deletion of the region of p300 encoding the histone acetyltransferase activity abolished its effect on GATA-4-dependent transcriptional activity. These results establish that p300 acts as a GATA-4 coactivator and that the p300 histone acetyltransferase activity is necessary for the functional interaction. PMID- 11481323 TI - Molecular cloning and characterization of PELP1, a novel human coregulator of estrogen receptor alpha. AB - Nuclear hormone receptors (NRs) are transcription factors whose activity is regulated by ligands and by coactivators or corepressors. We report the characterization of a new NR coregulator: proline-, glutamic acid-, leucine-rich protein 1 (PELP1), a novel human protein that comprises 1,282 amino acids and is localized on chromosome 17. The primary structure of PELP1 consists of several motifs present in most transcriptional regulators including nine NR-interacting boxes (LXXLL motifs), a zinc finger, and glutamic acid- and proline-rich regions. We demonstrate that PELP1 is a coactivator of estrogen receptor alpha (ERalpha). PELP1 enhances 17beta-estradiol-dependent transcriptional activation from the estrogen response element in a dose-dependent manner. PELP1 interacts with ERalpha and also with general transcriptional coactivators p300 and cAMP response element-binding protein-binding protein. PELP1 was differentially expressed in various human and murine tissues with the highest expression levels in the testes, mammary glands, and brain. We also provide evidence supporting the developmental regulation of PELP1 expression in murine mammary glands, the detectable expression of PELP1 in human mammary cancer cell lines, and the enhanced expression of PELP1 in human breast tumors. These findings suggest that PELP1 is a novel coregulator of ERalpha and may have a role in breast cancer tumorigenesis. PMID- 11481324 TI - Insulin receptor substrate-2 phosphorylation is necessary for protein kinase C zeta activation by insulin in L6hIR cells. AB - We have investigated glycogen synthase (GS) activation in L6hIR cells expressing a peptide corresponding to the kinase regulatory loop binding domain of insulin receptor substrate-2 (IRS-2) (KRLB). In several clones of these cells (B2, F4), insulin-dependent binding of the KRLB to insulin receptors was accompanied by a block of IRS-2, but not IRS-1, phosphorylation, and insulin receptor binding. GS activation by insulin was also inhibited by >70% in these cells (p < 0.001). The impairment of GS activation was paralleled by a similarly sized inhibition of glycogen synthase kinase 3 alpha (GSK3 alpha) and GSK3 beta inactivation by insulin with no change in protein phosphatase 1 activity. PDK1 (a phosphatidylinositol trisphosphate-dependent kinase) and Akt/protein kinase B (PKB) activation by insulin showed no difference in B2, F4, and in control L6hIR cells. At variance, insulin did not activate PKC zeta in B2 and F4 cells. In L6hIR, inhibition of PKC zeta activity by either a PKC zeta antisense or a dominant negative mutant also reduced by 75% insulin inactivation of GSK3 alpha and -beta (p < 0.001) and insulin stimulation of GS (p < 0.002), similar to Akt/PKB inhibition. In L6hIR, insulin induced protein kinase C zeta (PKC zeta) co precipitation with GSK3 alpha and beta. PKC zeta also phosphorylated GSK3 alpha and -beta. Alone, these events did not significantly affect GSK3 alpha and -beta activities. Inhibition of PKC zeta activity, however, reduced Akt/PKB phosphorylation of the key serine sites on GSK3 alpha and -beta by >80% (p < 0.001) and prevented full GSK3 inactivation by insulin. Thus, IRS-2, not IRS-1, signals insulin activation of GS in the L6hIR skeletal muscle cells. In these cells, insulin inhibition of GSK3 alpha and -beta requires dual phosphorylation by both Akt/PKB and PKC zeta. PMID- 11481325 TI - Protein interaction studies of MAGP-1 with tropoelastin and fibrillin-1. AB - Elastic fibers consist primarily of an amorphous elastin core associated with microfibrils, 10-12 nm in diameter, containing fibrillins and microfibril associated glycoproteins (MAGPs). To investigate the interaction of MAGP-1 with tropoelastin and fibrillin-1, we expressed human MAGP-1 as a T7-tag fusion protein in Escherichia coli. Refolding of the purified protein produced a soluble form of MAGP-1 that displayed saturable binding to tropoelastin. Fragments of tropoelastin corresponding to the N-terminal, C-terminal, and central regions of the molecule were used to characterize the MAGP-1 binding site. Cleavage of tropoelastin with kallikrein, which cleaves after Arg(515) in the central region of the molecule, disrupted the interaction, suggesting that the separated N- and C-terminal fragments were insufficient to determine MAGP-1 binding to intact tropoelastin. In addition, no evidence of an interaction was observed between MAGP-1 and a tropoelastin construct consisting of domains 17-27 that brackets the kallikrein cleavage site, suggesting a complex mechanism of interaction between the two molecules. Binding of MAGP-1 was also tested with overlapping recombinant fibrillin-1 fragments. MAGP-1 bound to a region at the N terminus of fibrillin-1 in a calcium-dependent manner. In summary, these results suggest a model for the interaction of elastin with the microfibrillar scaffold. PMID- 11481326 TI - Identification of a mouse thiamine transporter gene as a direct transcriptional target for p53. AB - p53 tumor suppressor is a transcription factor that functions, in part, through many of its downstream target genes. We have identified a p53-inducible gene by performing mRNA differential display on IW32 murine erythroleukemia cells containing a temperature-sensitive p53 mutant allele, tsp53(Val-135). Sequence analysis of the full-length cDNA revealed its identity as the mouse homologue of the human thiamine transporter 1 (THTR-1). Induction of the mouse THTR-1 (mTHTR 1) mRNA was detectable as early as 1 h at 32.5 degrees C; upon shifting back to 38.5 degrees C, mTHTR-1 transcript was rapidly degraded with a half-life of less than 2 h. Elevation of mTHTR-1 expression was found in DNA damage-induced normal mouse embryonic fibroblast cells, but not in p53(-/-) mouse embryonic fibroblast cells, suggesting that mTHTR-1 induction was p53-dependent. A region within the first intron of the mTHTR-1 gene bound to p53 and conferred the p53-mediated transactivation. Furthermore, increased thiamine transporter activities were found in cells overexpressing mTHTR-1 and under conditions of DNA damage or p53 activation. Our findings indicate that p53 may be involved in maintaining thiamine homeostasis through transactivation of THTR-1. PMID- 11481327 TI - The anti-initial transcribed sequence, a portable sequence that impedes promoter escape, requires sigma70 for function. AB - The anti-sequence, a portable element extending from +1 to +15 of the transcript, is sufficient to prevent promoter escape from a variety of strong final sigma70 promoters. We show here that this sequence does not function with even the strongest final sigma32 promoter. Moreover, a particular class of substitutions in final sigma70 that disrupt interaction between Region 2.2 of final sigma70 and a coiled-coiled motif in the beta'-subunit of RNA polymerase antagonizes the function of the anti-element. This same group of mutants prevents lambdaQ mediated anti-termination at the lambdaP(R') promoter. At this promoter, interaction of final sigma70 with the non-template strand of the initial transcribed sequence (ITS) is required to promote the pause prerequisite for anti termination. These mutants prevent pausing because they are defective in this recognition event. By analogy, we suggest that interaction of final sigma70 with the non-template strand of the anti-ITS is required for function of this portable element, thus explaining why neither final sigma32 nor the Region 2.2 final sigma70 mutants mediate anti-function. Support for the analogy with the lambdaP(R') promoter comes from preliminary experiments suggesting that the anti ITS, like the lambdaP(R') ITS, is bipartite. PMID- 11481328 TI - Glucose-induced metabolic oscillations parallel those of Ca(2+) and insulin release in clonal insulin-secreting cells. A multiwell approach to oscillatory cell behavior. AB - Insulin secretion from glucose-stimulated pancreatic beta-cells is oscillatory, and this is thought to result from oscillations in glucose metabolism. One of the primary metabolic stimulus-secretion coupling factors is the ATP/ADP ratio, which can oscillate as a result of oscillations in glycolysis. Using a novel multiwell culture plate system, we examined oscillations in insulin release and the ATP/ADP ratio in the clonal insulin-secreting cell lines HIT T-15 and INS-1. Insulin secretion from HIT cells grown in multiwell plates oscillated with a period of 4 min, similar to that seen previously in perifusion experiments. Oscillations in the ATP/ADP ratio in cells grown under the same conditions also occurred with a period of 4 min, as did oscillations in [Ca(2+)](i) monitored by fluorescence microscopy. In INS-1 cells oscillations in insulin secretion, the ATP/ADP ratio, and [Ca(2+)](i) were also seen, but with a shorter period of about 1.5 min. These observations of oscillations in the ATP/ADP ratio are consistent with their proposed role in driving the oscillations in [Ca(2+)](i) and insulin secretion. Furthermore, these data show that, at least in the clonal beta-cell lines, cell contact or even circulatory connection is not necessary for synchronous oscillations induced by a rise in glucose. PMID- 11481329 TI - Nuclear targeting is required for hepatoma-derived growth factor-stimulated mitogenesis in vascular smooth muscle cells. AB - We recently identified hepatoma-derived growth factor (HDGF) as a nuclear targeted vascular smooth muscle cell (VSM) mitogen that is expressed in developing vascular lesions. In the present study, VSM in culture express endogenous HDGF only in the nucleus and target a green fluorescent protein (GFP) HDGF fusion to the nucleus. To define the features of the HDGF molecule that are essential for nuclear localization and mitogenic function, deletion and site directed mutagenesis were performed. Deletion analysis identified the carboxyl terminal half of HDGF to be responsible for nuclear targeting in VSM. Overexpression of tagged HDGF proteins with point mutations in the putative bipartite nuclear localization sequence in the carboxyl terminus demonstrated that single Lys --> Asn mutations randomized HDGF expression to both the nucleus and cytoplasm similar to the empty vector. Importantly, the Lys --> Asn mutation of all three lysines blocked nuclear entry. Point mutation of a p34(cdc2) kinase consensus motif within the nuclear localization sequence had no effect on nuclear targeting. Moreover, nuclear entry was essential for the HDGF mitogenic effect, as transfection with the triple Lys --> Asn mutant HA-HDGF significantly attenuated bromodeoxyuridine uptake when compared with transfection with wild type HA-HDGF. We conclude that HDGF contains a true bipartite nuclear localization sequence with all three lysines necessary for nuclear targeting. Nuclear targeting of HDGF is required for HDGF stimulation of DNA replication in VSM. PMID- 11481330 TI - Identification and characterization of novel mammalian neuropeptide FF-like peptides that attenuate morphine-induced antinociception. AB - The two mammalian neuropeptides NPFF and NPAF have been shown to have important roles in nociception, anxiety, learning and memory, and cardiovascular reflex. Two receptors (FF1 and FF2) have been molecularly identified for NPFF and NPAF. We have now characterized a novel gene designated NPVF that encodes two neuropeptides highly similar to NPFF. NPVF mRNA was detected specifically in a region between the dorsomedial and ventromedial hypothalamic nuclei. NPVF-derived peptides displayed higher affinity for FF1 than NPFF-derived peptides, but showed poor agonist activity for FF2. Following intracerebral ventricular administration, a NPVF-derived peptide blocked morphine-induced analgesia more potently than NPFF in both acute and inflammatory models of pain. In situ hybridization analysis revealed distinct expression patterns of FF1 and FF2 in the rat central nervous system. FF1 was broadly distributed, with the highest levels found in specific regions of the limbic system and the brainstem where NPVF-producing neurons were shown to project. FF2, in contrast, was mostly expressed in the spinal cord and some regions of the thalamus. These results indicate that the endogenous ligands for FF1 and FF2 are NPVF- and NPFF-derived peptides, respectively, and suggest that the NPVF/FF1 system may be an important part of endogenous anti-opioid mechanism. PMID- 11481331 TI - Identification of tyrosine phosphorylation sites on 3-phosphoinositide-dependent protein kinase-1 and their role in regulating kinase activity. AB - 3-Phosphoinositide-dependent protein kinase-1 (PDK1) plays a central role in signal transduction pathways that activate phosphoinositide 3-kinase. Despite its key role as an upstream activator of enzymes such as protein kinase B and p70 ribosomal protein S6 kinase, the regulatory mechanisms controlling PDK1 activity are poorly understood. PDK1 has been reported to be constitutively active in resting cells and not further activated by growth factor stimulation (Casamayor, A., Morrice, N. A., and Alessi, D. R. (1999) Biochem. J. 342, 287-292). Here, we report that PDK1 becomes tyrosine-phosphorylated and translocates to the plasma membrane in response to pervanadate and insulin. Following pervanadate treatment, PDK1 kinase activity increased 1.5- to 3-fold whereas the activity of PDK1 associated with the plasma membrane increased approximately 6-fold. The activity of PDK1 localized to the plasma membrane was also increased by insulin treatment. Three tyrosine phosphorylation sites of PDK1 (Tyr-9 and Tyr-373/376) were identified using in vivo labeling and mass spectrometry. Using site-directed mutants, we show that, although phosphorylation on Tyr-373/376 is important for PDK1 activity, phosphorylation on Tyr-9 has no effect on the activity of the kinase. Both of these residues can be phosphorylated by v-Src tyrosine kinase in vitro, and co-expression of v-Src leads to tyrosine phosphorylation and activation of PDK1. Thus, these data suggest that PDK1 activity is regulated by reversible phosphorylation, possibly by a member of the Src kinase family. PMID- 11481332 TI - Identification of a novel Rab11/25 binding domain present in Eferin and Rip proteins. AB - Rab11, a low molecular weight GTP-binding protein, has been shown to play a key role in a variety of cellular processes, including endosomal recycling, phagocytosis, and transport of secretory proteins from the trans-Golgi network. In this study we have described a novel Rab11 effector, EF-hands-containing Rab11 interacting protein (Eferin). In addition, we have identified a 20-amino acid domain that is present at the C terminus of Eferin and other Rab11/25-interacting proteins, such as Rip11 and nRip11. Using biochemical techniques we have demonstrated that this domain is necessary and sufficient for Rab11 binding in vitro and that it is required for localization of Rab11 effector proteins in vivo. The data suggest that various Rab effectors compete with each other for binding to Rab11/25 possibly accounting for the diversity of Rab11 functions. PMID- 11481333 TI - DGAT2 is a new diacylglycerol acyltransferase gene family: purification, cloning, and expression in insect cells of two polypeptides from Mortierella ramanniana with diacylglycerol acyltransferase activity. AB - Acyl CoA:diacylgycerol acyltransferase (EC; DGAT) catalyzes the final step in the production of triacylglycerol. Two polypeptides, which co-purified with DGAT activity, were isolated from the lipid bodies of the oleaginous fungus Mortierella ramanniana with a procedure consisting of dye affinity, hydroxyapatite affinity, and heparin chromatography. The two enzymes had molecular masses of 36 and 36.5 kDa, as estimated by gel electrophoresis, and showed a broad activity maximum between pH 6 and 8. Based on partial peptide sequence information, polymerase chain reaction techniques were used to obtain full-length cDNA sequences encoding the purified proteins. Expression of the cDNAs in insect cells conferred high levels of DGAT activity on the membranes isolated from these cells. The two proteins share 54% homology with each other but are unrelated to the previously identified DGAT gene family (designated DGAT1), which is related to the acyl CoA:cholesterol acyltransferase gene family, or to any other gene family with ascribed function. This report identifies a new gene family, including members in fungi, plants and animals, which encode enzymes with DGAT function. To distinguish the two unrelated families we designate this new class DGAT2 and refer to the M. ramanniana genes as MrDGAT2A and MrDGAT2B. PMID- 11481334 TI - Stereochemistry of quinoxaline antagonist binding to a glutamate receptor investigated by Fourier transform infrared spectroscopy. AB - The stereochemistry of the interactions between quinoxaline antagonists and the ligand-binding domain of the glutamate receptor 4 (GluR4) have been investigated by probing their vibrational modes using Fourier transform infrared spectroscopy. In solution, the electron-withdrawing nitro groups of both compounds establish a resonance equilibrium that appears to stabilize the keto form of one of the cyclic amide carbonyl bonds. Changes in the 6,7-dinitro-2,3-dihydroxyquinoxaline vibrational spectra on binding to the glutamate receptor, interpreted within the framework of a published crystal structure, illuminate the stereochemistry of the interaction and suggest that the binding site imposes a more polarized electronic bonding configuration on this antagonist. Similar spectral changes are observed for 6-cyano-7-dinitro-2,3-dihydroxyquinoxaline, confirming that its interactions with the binding site are highly similar to those of 6,7-dinitro-2,3 dihydroxyquinoxaline and leading to a model of the 6-cyano-7-dinitro-2,3 dihydroxyquinoxaline-S1S2 complex, for which no crystal structure is available. Conformational changes within the GluR ligand binding domain were also monitored. Compared with the previously reported spectral changes seen on binding of the agonist glutamate, only a relatively small change is detected on antagonist binding. This correlation between the functional effects of different classes of ligand and the magnitude of the spectroscopic changes they induce suggests that the spectral data reflect physiologically relevant conformational processes. PMID- 11481335 TI - Cloning of DGAT2, a second mammalian diacylglycerol acyltransferase, and related family members. AB - Studies involving the cloning and disruption of the gene for acyl CoA:diacylglycerol acyltransferase (DGAT) have shown that alternative mechanisms exist for triglyceride synthesis. In this study, we cloned and characterized a second mammalian DGAT, DGAT2, which was identified by its homology to a DGAT in the fungus Mortierella rammaniana. DGAT2 is a member of a gene family that has no homology with DGAT1 and includes several mouse and human homologues that are candidates for additional DGAT genes. The expression of DGAT2 in insect cells stimulated triglyceride synthesis 6-fold in assays with cellular membranes, and DGAT2 activity was dependent on the presence of fatty acyl-CoA and diacylglycerol, indicating that this protein is a DGAT. Activity was not observed for acyl acceptors other than diacylglycerol. DGAT2 activity was inhibited by a high concentration (100 mm) of MgCl(2) in an in vitro assay, a characteristic that distinguishes DGAT2 from DGAT1. DGAT2 is expressed in many tissues with high expression levels in the liver and white adipose tissue, suggesting that it may play a significant role in mammalian triglyceride metabolism. PMID- 11481336 TI - Genetic and physical interactions between Microphthalmia transcription factor and PU.1 are necessary for osteoclast gene expression and differentiation. AB - The microphthalmia transcription factor (MITF), a basic-helix-loop-helix zipper factor, regulates distinct target genes in several cell types. We hypothesized that interaction with the Ets family factor PU.1, whose expression is limited to hematopoietic cells, might be necessary for activation of target genes like tartrate-resistant acid phosphatase (TRAP) in osteoclasts. Several lines of evidence were consistent with this model. The combination of MITF and PU.1 synergistically activated the TRAP promoter in transient assays. This activation was dependent on intact binding sites for both factors in the TRAP promoter. MITF and PU.1 physically interacted when coexpressed in COS cells or in vitro when purified recombinant proteins were studied. The minimal regions of MITF and PU.1 required for the interaction were the basic-helix-loop-helix zipper domain and the Ets DNA binding domain, respectively. Significantly, mice heterozygous for both the mutant mi allele and a PU.1 null allele developed osteopetrosis early in life which resolved with age. The size and number of osteoclasts were not altered in the double heterozygous mutant mice, indicating that the defect lies in mature osteoclast function. Taken in total, the results afford an example of how lineage specific gene regulation can be achieved by the combinatorial action of two broadly expressed transcription factors. PMID- 11481337 TI - Lipoprotein secretion and triglyceride stores in the heart. AB - The genes for apolipoprotein B and microsomal triglyceride transfer protein are expressed in mouse and human heart tissue. Why the heart would express these "lipoprotein assembly" genes has been unclear. Here we demonstrate that the beating mouse heart actually secretes spherical lipoproteins. Moreover, increased cardiac production of lipoproteins (e.g., in mice that express a human apolipoprotein B transgene) was associated with increased triglyceride secretion from the heart and decreased stores of triglycerides within the heart. Increased cardiac production of lipoproteins also reduced the pathological accumulation of triglycerides that occurs in the hearts of mice lacking long-chain acyl coenzyme A dehydrogenase. In contrast, blocking heart lipoprotein secretion (e.g., in heart-specific microsomal triglyceride transfer protein knockout mice) increased cardiac triglyceride stores. Thus, heart lipoprotein secretion helps regulate cardiac triglyceride stores and may protect the heart from the detrimental effects of surplus lipids. PMID- 11481338 TI - Identification of the human methylmalonyl-CoA racemase gene based on the analysis of prokaryotic gene arrangements. Implications for decoding the human genome. AB - In this report, we identify the human DL-methylmalonyl-CoA racemase gene by analyzing prokaryotic gene arrangements and extrapolating the information obtained to human genes by homology searches. Sequence similarity searches were used to identify two groups of homologues that were frequently arranged with prokaryotic methylmalonyl-CoA mutase genes, and that were of unknown function. Both gene groups had homologues in the human genome. Because methylmalonyl-CoA mutases are involved in the metabolism of propionyl-CoA, we inferred that conserved neighbors of methylmalonyl-CoA mutase genes and their human homologues were also involved in this process. Subsequent biochemical studies confirmed this inference by showing that the prokaryotic gene PH0272 and its human homologue both encode DL-methylmalonyl-CoA racemases. To our knowledge this is the first report in which the function of a eukaryotic gene was determined based on the analysis of prokaryotic gene arrangements. Importantly, such analyses are rapid and may be generally applicable for the identification of human genes that lack homologues of known function or that have been misidentified on the basis of sequence similarity searches. PMID- 11481344 TI - Clathrin-dependent and -independent internalization of plasma membrane sphingolipids initiates two Golgi targeting pathways. AB - Sphingolipids (SLs) are plasma membrane constituents in eukaryotic cells which play important roles in a wide variety of cellular functions. However, little is known about the mechanisms of their internalization from the plasma membrane or subsequent intracellular targeting. We have begun to study these issues in human skin fibroblasts using fluorescent SL analogues. Using selective endocytic inhibitors and dominant negative constructs of dynamin and epidermal growth factor receptor pathway substrate clone 15, we found that analogues of lactosylceramide and globoside were internalized almost exclusively by a clathrin independent ("caveolar-like") mechanism, whereas an analogue of sphingomyelin was taken up approximately equally by clathrin-dependent and -independent pathways. We also showed that the Golgi targeting of SL analogues internalized via the caveolar-like pathway was selectively perturbed by elevated intracellular cholesterol, demonstrating the existence of two discrete Golgi targeting pathways. Studies using SL-binding toxins internalized via clathrin-dependent or independent mechanisms confirmed that endogenous SLs follow the same two pathways. These findings (a) provide a direct demonstration of differential SLs sorting into early endosomes in living cells, (b) provide a "vital marker" for endosomes derived from caveolar-like endocytosis, and (c) identify two independent pathways for lipid transport from the plasma membrane to the Golgi apparatus in human skin fibroblasts. PMID- 11481345 TI - Activation of ARF6 by ARNO stimulates epithelial cell migration through downstream activation of both Rac1 and phospholipase D. AB - Migration of epithelial cells is essential for tissue morphogenesis, wound healing, and metastasis of epithelial tumors. Here we show that ARNO, a guanine nucleotide exchange factor for ADP-ribosylation factor (ARF) GTPases, induces Madin-Darby canine kidney epithelial cells to develop broad lamellipodia, to separate from neighboring cells, and to exhibit a dramatic increase in migratory behavior. This transition requires ARNO catalytic activity, which we show leads to enhanced activation of endogenous ARF6, but not ARF1, using a novel pulldown assay. We further demonstrate that expression of ARNO leads to increased activation of endogenous Rac1, and that Rac activation is required for ARNO induced cell motility. Finally, ARNO-induced activation of ARF6 also results in increased activation of phospholipase D (PLD), and inhibition of PLD activity also inhibits motility. However, inhibition of PLD does not prevent activation of Rac. Together, these data suggest that ARF6 activation stimulates two distinct signaling pathways, one leading to Rac activation, the other to changes in membrane phospholipid composition, and that both pathways are required for cell motility. PMID- 11481346 TI - Barentsz is essential for the posterior localization of oskar mRNA and colocalizes with it to the posterior pole. AB - The localization of Oskar at the posterior pole of the Drosophila oocyte induces the assembly of the pole plasm and therefore defines where the abdomen and germ cells form in the embryo. This localization is achieved by the targeting of oskar mRNA to the posterior and the localized activation of its translation. oskar mRNA seems likely to be actively transported along microtubules, since its localization requires both an intact microtubule cytoskeleton and the plus end directed motor kinesin I, but nothing is known about how the RNA is coupled to the motor. Here, we describe barentsz, a novel gene required for the localization of oskar mRNA. In contrast to all other mutations that disrupt this process, barentsz-null mutants completely block the posterior localization of oskar mRNA without affecting bicoid and gurken mRNA localization, the organization of the microtubules, or subsequent steps in pole plasm assembly. Surprisingly, most mutant embryos still form an abdomen, indicating that oskar mRNA localization is partially redundant with the translational control. Barentsz protein colocalizes to the posterior with oskar mRNA, and this localization is oskar mRNA dependent. Thus, Barentsz is essential for the posterior localization of oskar mRNA and behaves as a specific component of the oskar RNA transport complex. PMID- 11481347 TI - Myosin light chain kinase binding to a unique site on F-actin revealed by three dimensional image reconstruction. AB - Ca2+-calmodulin-dependent phosphorylation of myosin regulatory light chains by the catalytic COOH-terminal half of myosin light chain kinase (MLCK) activates myosin II in smooth and nonmuscle cells. In addition, MLCK binds to thin filaments in situ and F-actin in vitro via a specific repeat motif in its NH2 terminus at a stoichiometry of one MLCK per three actin monomers. We have investigated the structural basis of MLCK-actin interactions by negative staining and helical reconstruction. F-actin was decorated with a peptide containing the NH2-terminal 147 residues of MLCK (MLCK-147) that binds to F-actin with high affinity. MLCK-147 caused formation of F-actin rafts, and single filaments within rafts were used for structural analysis. Three-dimensional reconstructions showed MLCK density on the extreme periphery of subdomain-1 of each actin monomer forming a bridge to the periphery of subdomain-4 of the azimuthally adjacent actin. Fitting the reconstruction to the atomic model of F-actin revealed interaction of MLCK-147 close to the COOH terminus of the first actin and near residues 228-232 of the second. This unique location enables MLCK to bind to actin without interfering with the binding of any other key actin-binding proteins, including myosin, tropomyosin, caldesmon, and calponin. PMID- 11481348 TI - Matrix metalloproteinase inhibition of pancreatic cancer: matching mechanism of action to clinical trial design. PMID- 11481349 TI - Marimastat as first-line therapy for patients with unresectable pancreatic cancer: a randomized trial. AB - PURPOSE: The prognosis for unresectable pancreatic cancer remains dismal (1-year survival rate, < 10%; 5-year survival rate, < 5%). Recent advances in conventional chemotherapy and novel molecular treatment strategies warrant investigation. This, the largest randomized study in pancreatic cancer performed to date, compares marimastat, the first of a new class of agents, with gemcitabine. PATIENTS AND METHODS: Four hundred fourteen patients with unresectable pancreatic cancer were randomized to receive marimastat 5, 10, or 25 mg bid or gemcitabine 1,000 mg/m2. The primary end point was survival. Progression-free survival, patient benefit, and safety were also assessed. RESULTS: There was no significant difference in survival between 5, 10, or 25 mg of marimastat and gemcitabine (P =.19). Median survival times were 111, 105, 125, and 167 days, respectively, and 1-year survival rates were 14%, 14%, 20%, and 19%, respectively. There was a significant difference in survival rates between patients treated with gemcitabine and marimastat 5 and 10 mg (P <.003). Both agents were well tolerated, although grade 3 or 4 toxicities were reported in 22% and 12% of the gemcitabine- and marimastat-treated patients, respectively. The major toxicity of marimastat was musculoskeletal (44% of marimastat patients, compared with 12% of gemcitabine patients; musculoskeletal toxicity was severe in only 8% of marimastat patients). CONCLUSION: The results of this study provide evidence of a dose response for marimastat in patients with advanced pancreatic cancer. The 1-year survival rate for patients receiving marimastat 25 mg was similar to that of patients receiving gemcitabine. In view of the manageable tolerability of marimastat and its ease of administration, further studies are warranted. PMID- 11481350 TI - Sequence effect of irinotecan and fluorouracil treatment on pharmacokinetics and toxicity in chemotherapy-naive metastatic colorectal cancer patients. AB - PURPOSE: To investigate the sequence effect of irinotecan and a 48-hour infusion of fluorouracil (5-FU) modulated by leucovorin (LV) on the plasma pharmacokinetics of irinotecan and its metabolites, the toxicity profile of this combination, and irinotecan's maximum-tolerated dose (MTD). PATIENTS AND METHODS: Thirty-three metastatic colorectal cancer patients were randomized to receive a 60-minute infusion of irinotecan before or after a 48-hour infusion of 5-FU modulated by LV. The reverse sequence was used after 21 days for the second cycle. 5-FU 3,500 mg/m2 was preceded by l-LV 250 mg/m2. Irinotecan 150 mg/m2 (starting dose) was administered to the first three patients. The dose was escalated by 50 mg/m2 in subsequent groups of three to six patients to determine the MTD for both sequences. Pharmacokinetic analysis of irinotecan and its metabolites was performed after each cycle. RESULTS: Toxicities were affected by the sequence of administration of irinotecan and 5-FU, with an improved tolerability for irinotecan followed by 5-FU. The irinotecan MTD was reached at 300 mg/m2 when irinotecan followed 5-FU and at 450 mg/m2 when it preceded 5-FU. In seven of 23 patients who received both sequences at identical irinotecan doses, the dose-limiting toxicity was observed only when irinotecan followed 5 FU. Pharmacokinetic analysis revealed that the administration sequence significantly affected the SN-38 area under the concentration-versus-time curve (AUC), which was 40.1% lower (P <.05) when irinotecan preceded 5-FU. CONCLUSION: The sequence of treatment with irinotecan and infusional 5-FU affects the tolerability of this combination. This can be explained in part by a reduced SN 38 AUC when irinotecan preceded infusional 5-FU. Well-defined 5-FU/irinotecan regimens are needed because the administration sequence or the interval between the agents might affect treatment tolerance and perhaps also activity. PMID- 11481351 TI - Cyclophosphamide plus topotecan in children with recurrent or refractory solid tumors: a Pediatric Oncology Group phase II study. AB - PURPOSE: To determine the response rate of the combination of cyclophosphamide and topotecan in pediatric patients with recurrent or refractory malignant solid tumors. PATIENTS AND METHODS: A total of 91 pediatric patients, 83 of whom were fully assessable for response and toxicity, received cyclophosphamide (250 mg/m2/dose) followed by topotecan (0.75 mg/m2/dose), each given as a 30-minute infusion daily for 5 days. All patients received filgrastim (5 mcg/kg) daily until the absolute neutrophil count (ANC) was > or = 1,500 microL after the time of the expected ANC nadir. RESULTS: A total of 307 treatment courses were given to the 83 fully assessable patients. Responses (complete response plus partial response) were seen in rhabdomyosarcoma (10 of 15 patients), Ewing's sarcoma (six of 17 patients), and neuroblastoma (six of 13 patients). Partial responses were seen in two of 18 patients with osteosarcoma and in one patient with a Sertoli Leydig cell tumor. Twenty-three patients had either minor responses (n = 6) or stable disease (n = 17); the median number of courses administered to patients with partial or complete response was six (range, two to 13 courses), and the median administered to those with stable disease was three (range, one to 11 courses). The toxicity of the combination was limited principally to the hematopoietic system. Of 307 courses, 163 (53%) were associated with grade 3 or 4 neutropenia, 84 (27%) with grade 3 or 4 anemia, and 136 (44%) with grade 3 or 4 thrombocytopenia. Despite the severe myelosuppression, only 34 (11%) of 307 courses were associated with grade 3 or 4 infection. Nonhematopoietic toxicity of grades > or = 3 was rare and consisted of nausea and vomiting (two courses), perirectal mucositis (one course), transaminase elevation (one course), and hematuria (two courses). CONCLUSION: The combination of cyclophosphamide and topotecan is active in rhabdomyosarcoma, neuroblastoma, and Ewing's sarcoma. Stabilization of disease was seen in osteosarcoma, although objective responses were rare in this disease. The therapy can be given with acceptable hematopoietic toxicity with the use of filgrastim support. PMID- 11481352 TI - Intellectual outcome after reduced-dose radiation therapy plus adjuvant chemotherapy for medulloblastoma: a Children's Cancer Group study. AB - PURPOSE: To investigate the intellectual outcomes of children with medulloblastomas/primitive neuroectodermal tumors (MB/PNET) treated with reduced dose craniospinal radiotherapy (RT) plus adjuvant chemotherapy. PATIENTS AND METHODS: Forty-three children with average-risk posterior fossa MB/PNETs underwent longitudinal intelligence testing. All had been treated with a reduced dose craniospinal RT regimen (23.4 Gy to the neuraxis, 32.4-Gy boost to the posterior fossa) and adjuvant chemotherapy. RESULTS: The estimated rate of change from baseline was significant for Full Scale Intelligence Quotient (FSIQ), Verbal IQ (VIQ), and Nonverbal IQ (NVIQ) (P <.001 for all three outcomes). The rate of change was estimated to be -4.3 FSIQ points per year, -4.2 VIQ points per year, and -4.0 NVIQ points per year. Females were more subject to VIQ decline than were males (P =.008), and young children (< 7 years of age) were more negatively affected than were older children, with a significant decline in NVIQ (P =.016). Finally, patients with higher baseline evaluations suffered greater declines in IQ than did those with lower baseline scores. CONCLUSION: This study represents the largest series of patients with average-risk MB/PNETs treated with a combination of reduced-dose RT and adjuvant chemotherapy whose intellectual development has been followed prospectively. Intellectual loss was substantial but suggestive of some degree of intellectual preservation compared with effects associated with conventional RT doses. However, this conclusion remains provisional, pending further research. PMID- 11481353 TI - Factors associated with response to high-dose interleukin-2 in patients with metastatic melanoma. AB - PURPOSE: The present study attempted to identify characteristics that correlated with clinical response to interleukin (IL)-2 therapy in patients with metastatic melanoma. PATIENTS AND METHODS: We retrospectively evaluated laboratory and clinical characteristics of 374 consecutive patients with metastatic melanoma treated with high-dose intravenous bolus IL-2 (720,000 IU/kg) from July 1, 1988, to December 31, 1999, at the Surgery Branch of the National Cancer Institute. RESULTS: The overall objective response rate was 15.5%. Pretreatment parameters such as patient demographics, laboratory values, and prior therapy did not correlate with response; however, 53.6% of patients with only subcutaneous and/or cutaneous metastases responded, compared with 12.4% of patients with disease at other sites (P2 =.000001). During therapy, patients who were responders tended to have received more doses during course 1 (16.2 +/- 0.3 doses v 14.5 +/- 0.2 doses; P2 =.0095); however, when limited to patients who were able to complete both cycles of course 1, there was no statistically significant difference (P2 =.27). Responders had a higher maximum lymphocyte count immediately after therapy compared with nonresponders (P2 =.0026). The development of abnormal thyroid function tests and vitiligo after therapy was associated with response (thyroid stimulating hormone, P2 =.01; free T4, P2 =.0049; vitiligo, P2 < 10(-6)), although thyroid dysfunction may have been related more to the length of IL-2 therapy than to response. CONCLUSION: The presence of metastases only to subcutaneous and/or cutaneous sites, lymphocytosis immediately after treatment, and long-term immunologic side effects, especially vitiligo, were associated with antitumor response to IL-2 therapy. PMID- 11481354 TI - Phase II clinical investigation of gemcitabine in advanced soft tissue sarcomas and window evaluation of dose rate on gemcitabine triphosphate accumulation. AB - PURPOSE: To evaluate the efficacy, toxicity, and optimal dose rate of gemcitabine in adult patients with advanced soft tissue sarcomas (STS) by comparing levels of gemcitabine triphosphate (GTP) in peripheral-blood mononuclear cells (PBMCs) of patients receiving two different dose rates. PATIENTS AND METHODS: Fifty-six assessable patients with STS (17 gastrointestinal [GI] leiomyosarcomas and 39 other histologies) were treated on a two-arm phase II study. Gemcitabine was given at 1 g/m2 as a 30-minute infusion weekly for up to 7 weeks followed by 1 week of rest and reassessment of tumor. Subsequent cycles were given at 1 g/m2 weekly for 3 weeks followed by 1 week of rest. Nine patients underwent cellular pharmacologic studies at two different dose rates (1 g/m2 over a standard 30 minute infusion on week 1 and over pharmacologically based infusion of 150 minutes on week 2) to evaluate GTP levels in PBMCs. RESULTS: Seven partial responses were noted among 39 patients, for an overall response rate of 18% (95% confidence interval, 7% to 29%). Median duration of response was 3.5 months (range, 2 to 13 months). Four of 10 patients with non-GI leiomyosarcomas achieved a partial response. No objective responses were noted in 17 patients with GI leiomyosarcomas. One patient had a mixed response. Median time to progression for all patients (both arms) was 3 months; median survival was 13.9 months. Treatment was generally well tolerated. Comparison of cellular pharmacology demonstrated a significant 1.4-fold increase in the concentration of GTP with the 150-minute infusion. CONCLUSION: Given the limited therapeutic armamentarium for STS, the activity of gemcitabine is encouraging. Its potential for combination therapy in the salvage setting should be studied with pharmacologically guided fixed dose rate infusion. PMID- 11481355 TI - Screening sensitivity and sojourn time from breast cancer early detection clinical trials: mammograms and physical examinations. AB - PURPOSE: To estimate sensitivities of breast cancer screening modalities and preclinical duration of the disease from eight breast cancer screening clinical trials. PATIENTS AND METHODS: Screening programs invariably lead to diagnosis of disease before signs or symptoms are present. Two key quantities of screening programs are the sensitivity of the disease detection modality and the mean sojourn time (MST). The observed screening histories in a periodically screened cohort make it possible to estimate these quantities of interest. We applied recently developed statistical methods to data from eight randomized breast cancer screening trials to estimate the sensitivities of early detection modalities and MST. Moreover, when a screening trial involved two screening modalities, our methods enabled the estimation of the individual sensitivity of each screening modality. RESULTS: We analyzed breast cancer data from several screening trials and have relatively complete data from the Health Insurance Plan (HIP), Edinburgh, and two Canadian studies. The screening sensitivity for mammography, physical examination, and MST were, respectively, HIP: 0.39, 0.47, and 2.5 years; Edinburgh: 0.63, 0.40, and 4.3 years; Canadian (age 40 to 49 at entry): 0.61, 0.59, and 1.9 years; Canadian (age 50 to 59 at entry): 0.66, 0.39, and 3.1 years. CONCLUSION: The public debate on early breast cancer detection is mainly centered on mammograms. However, the current study indicates that a physical examination is of comparable importance. Cautious interpretation of trial differences is required as a result of various experimental designs and the age dependency of screening sensitivity and MST. PMID- 11481356 TI - Weekly docetaxel in the treatment of elderly patients with advanced breast cancer: a Minnie Pearl Cancer Research Network phase II trial. AB - PURPOSE: To evaluate the efficacy and toxicity of docetaxel administered weekly to elderly or poor-performance status patients with advanced breast cancer. PATIENTS AND METHODS: Forty-one patients with advanced breast cancer who were either over the age of 65 or considered to be poor candidates for combination chemotherapy received docetaxel 36 mg/m2 weekly for 6 consecutive weeks, followed by 2 weeks without treatment. The median age of patients in this trial was 74 years, and 73% of patients had one or more visceral sites of metastases. Seventy five percent of patients received weekly docetaxel as first-line treatment for metastatic breast cancer, and the other 25% received it as second-line treatment. Thirty-six patients were assessable for efficacy, and all patients were assessed for toxicity. RESULTS: A total of 448 doses of weekly docetaxel were administered to 41 patients. Thirteen patients (36%) had objective responses to treatment, and an additional 13 patients (36%) had stable disease or minor response. Median time to progression for responding and stable patients was 7 months (range, 3 to 27 months). Median survival for the entire group was 13 months, with 1- and 2-year actuarial survival rates of 61% and 29%, respectively. Severe neutropenia occurred in only 0.4% of courses, and no other hematologic toxicity was observed. Grade 3/4 fatigue was the most common toxicity, occurring in 20% of patients. CONCLUSION: Weekly docetaxel therapy is active and well tolerated by elderly and/or poor-performance status patients with advanced breast cancer. This treatment can be administered with minimal myelosuppression. Weekly docetaxel provides an additional option for treatment in this difficult subgroup of patients with metastatic breast cancer. Well-tolerated combination regimens containing weekly docetaxel merit evaluation for this patient population. PMID- 11481357 TI - Dose-dense doxorubicin, docetaxel, and granulocyte colony-stimulating factor support with or without tamoxifen as preoperative therapy in patients with operable carcinoma of the breast: a randomized, controlled, open phase IIb study. AB - PURPOSE: To investigate the effect of adding tamoxifen to a preoperative dose dense doxorubicin and docetaxel regimen on the pathologic response of primary operable breast cancer. PATIENTS AND METHODS: Patients (tumor size > or = 3 cm, N0 to 2, M0) were prospectively randomized to receive every 14 days a total of four cycles of doxorubicin 50 mg/m2 and docetaxel 75 mg/m(2), either with (ADocT) or without (ADoc) simultaneous tamoxifen. Granulocyte colony-stimulating factor (G-CSF) was routinely given on days 5 to 10. Surgery followed 8 to 10 weeks after the start of treatment. RESULTS: Within 14 months, 250 patients were included in the study at 56 centers. Of 992 planned cycles, 97.9% were administered. Pathologically complete remission (pCR) with no detectable viable tumor cells was achieved in 9.7%. There was a nonsignificant difference of -1.2% in favor of ADoc, with a 95% confidence interval of -8.6% to 6.2%. A further 2.4% had only noninvasive tumor residues, and 13.8% had focal invasive residues. Complete and partial responses detected by palpation were observed in 28.9% and 52.4%, respectively. The response rates (complete and partial) by best appropriate imaging methods were 77.5% and 67.5% for ADocT and ADoc, respectively. Breast conservation was possible in 68.8% of the patients. A tendency toward more frequent toxic events was observed with ADocT treatment. Significant predictors of pCR to chemotherapy were negative lymph node and negative estrogen receptor status. CONCLUSION: A dose-dense regimen of ADoc with G-CSF offers high compliance, moderate toxicity, and rapid efficacy as a form of preoperative chemotherapy in operable breast cancer. Concurrent treatment with tamoxifen for 8 weeks could not improve the pathologic response rate. PMID- 11481358 TI - 18fluorodeoxyglucose positron emission tomography to detect mediastinal or internal mammary metastases in breast cancer. AB - PURPOSE: To determine the prevalence of suspected disease in the mediastinum and internal mammary (IM) node chain by 18fluorodeoxyglucose (FDG) positron emission tomography (PET), compared with conventional staging by computed tomography (CT) in patients with recurrent or metastatic breast cancer. PATIENTS AND METHODS: We retrospectively evaluated intrathoracic lymph nodes using FDG PET and CT data in 73 consecutive patients with recurrent or metastatic breast cancer who had both CT and FDG PET within 30 days of each other. In reviews of CT scans, mediastinal nodes measuring 1 cm or greater in the short axis were considered positive. PET was considered positive when there were one or more mediastinal foci of FDG uptake greater than the mediastinal blood pool. RESULTS: Overall, 40% of patients had abnormal mediastinal or IM FDG uptake consistent with metastases, compared with 23% of patients who had suspiciously enlarged mediastinal or IM nodes by CT. Both FDG PET and CT were positive in 22%. In the subset of 33 patients with assessable follow-up by CT or biopsy, the sensitivity, specificity, and accuracy for nodal disease was 85%, 90%, and 88%, respectively, by FDG PET; 54%, 85%, and 73%, respectively, by prospective interpretation of CT; and 50%, 83%, and 70%, respectively, by blinded observer interpretation of CT. Among patients suspected of having only locoregional disease recurrence (n = 33), 10 had unsuspected mediastinal or IM disease by FDG PET. CONCLUSION: FDG PET may uncover disease in these nodal regions not recognized by conventional staging methods. Future prospective studies using histopathology for confirmation are needed to validate the preliminary findings of this retrospective study. PMID- 11481359 TI - Comparison of breast magnetic resonance imaging, mammography, and ultrasound for surveillance of women at high risk for hereditary breast cancer. AB - PURPOSE: Recommended surveillance for BRCA1 and BRCA2 mutation carriers includes regular mammography and clinical breast examination, although the effectiveness of these screening techniques in mutation carriers has not been established. The purpose of the present study was to compare breast magnetic resonance imaging (MRI) with ultrasound, mammography, and physical examination in women at high risk for hereditary breast cancer. PATIENTS AND METHODS: A total of 196 women, aged 26 to 59 years, with proven BRCA1 or BRCA2 mutations or strong family histories of breast or ovarian cancer underwent mammography, ultrasound, MRI, and clinical breast examination on a single day. A biopsy was performed when any of the four investigations was judged to be suspicious for malignancy. RESULTS: Six invasive breast cancers and one noninvasive breast cancer were detected among the 196 high-risk women. Five of the invasive cancers occurred in mutation carriers, and the sixth occurred in a woman with a previous history of breast cancer. The prevalence of invasive or noninvasive breast cancer in the 96 mutation carriers was 6.2%. All six invasive cancers were detected by MRI, all were 1.0 cm or less in diameter, and all were node-negative. In contrast, only three invasive cancers were detected by ultrasound, two by mammography, and two by physical examination. The addition of MRI to the more commonly available triad of mammography, ultrasound, and breast examination identified two additional invasive breast cancers that would otherwise have been missed. CONCLUSION: Breast MRI may be superior to mammography and ultrasound for the screening of women at high risk for hereditary breast cancer. PMID- 11481360 TI - Multicenter phase II trial of paclitaxel, cisplatin, and etoposide with concurrent radiation for limited-stage small-cell lung cancer. AB - PURPOSE: To investigate the feasibility, efficacy, and safety of adding paclitaxel to cisplatin/etoposide chemotherapy and concurrent thoracic radiotherapy (TRT) in treatment of limited-stage small-cell lung cancer (LD SCLC). PATIENTS AND METHODS: Patients received five courses of chemotherapy (paclitaxel 175 mg/m2 1-hour intravenous [IV] infusion day 1; cisplatin 50 mg/m(2) IV day 1; etoposide 100 mg/m2 IV day 1; oral etoposide 100 mg bid days 2 to 5) at 3-week intervals. TRT (42 Gy administered in 15 fractions) was administered concurrent with chemotherapy cycle 3. All patients were evaluated before starting TRT and 4 weeks after termination of chemotherapy. Patients achieving complete remission (CR) were administered prophylactic cranial irradiation. RESULTS: Thirty-nine patients were included, and the median age was 63 years. The median follow-up was 36 months (range, 19 to 57 months). The overall response rate was 92% (CR, 81%; partial response, 11%), and the median survival was 21 months. The 1- and 2-year disease-specific survival rates were 69% and 37%, respectively. Of 29 CR patients, 83% have relapsed. Brain metastasis was as frequent as local recurrences (42%). Hematologic toxicity included grade 3 to 4 leukopenia in 74% of patients and grade 3 thrombocytopenia in 10%. One treatment-related death occurred as a result of severe neutropenia and septicemia. Hematotoxicity caused dose reductions in 31% of courses. One patient had an anaphylactic reaction during the first paclitaxel infusion. Paclitaxel related neuropathy and myalgia were reversible. Grade 3 esophagitis was seen in five patients during and shortly after TRT. CONCLUSION: This novel multimodal regimen is effective and well tolerated in patients with LD-SCLC. It compares favorably with previously published phase II studies. PMID- 11481361 TI - A simple stratification factor prognostic for survival in advanced cancer: the good/bad/uncertain index. AB - PURPOSE: This article summarizes the third step of a research program to identify variables that supplement the predictive power of the the Eastern Cooperative Oncology Group (ECOG) performance status (PS) for survival. The objective was to produce a simple, practical, stratification factor for phase III oncology clinical trials involving patients with advanced malignant disease. PATIENTS AND METHODS: A questionnaire was administered to 729 patients with metastatic colorectal or lung cancers. Patients provided a Karnofsky index and appetite rating while physicians provided a survival estimate and the ECOG-PS. Scores for each item were categorized as having a positive, neutral, or negative indication for survival. A patient was classified as having a relatively good prognosis if three or more of the four items showed a positive indication, a bad prognosis if three or more items were negative, and an uncertain prognosis otherwise (Good/Bad/Uncertain [GBU] index). RESULTS: The GBU index improved on the prognostic power of a Cox model quartile index and PS alone and increased the accuracy of survival classification estimates by 5% to 10% more than ECOG-PS alone. For patients with PS of 0 or 1, significant survival patterns exist between GBU groups (P=.002 and.0001, respectively). CONCLUSION: The GBU index may be recommended as a supplementary stratification factor for certain future phase III trials in metastatic lung or colorectal cancer where patient heterogeneity is a particular concern. The GBU represents a relatively modest increase to the cost and patient burden of a clinical trial given the additional control that is achieved over the potentially confounding concomitant to the treatment variable. PMID- 11481362 TI - High efficacy of fractionated stereotactic radiotherapy of large base-of-skull meningiomas: long-term results. AB - PURPOSE: Large skull-base meningiomas are difficult to treat due to their proximity or adherence to critical structures. We analyzed the long-term results of patients with skull-base meningiomas treated by a new approach with high precision fractionated stereotactic radiotherapy. PATIENTS AND METHODS: One hundred eighty-nine patients with benign meningiomas were treated with conformal fractionated stereotactic radiotherapy between 1985 and 1998. Patients were undergoing a course of radiotherapy either as primary treatment, following subtotal resection, or for recurrent disease. The median target volume was 52.5 mL (range, 5.2 to 370 mL). The mean radiation dose was 56.8 Gy (+/- 4.4 Gy). Follow-up examinations, including magnetic resonance imaging, were performed at 6 month intervals thereafter. RESULTS: The median follow-up period was 35 months (range, 3 months to 12 years). Overall actuarial survival for patients with World Health Organization (WHO) grade I meningiomas was 97% after 5 years and 96% after 10 years. Local tumor failure was observed in three of 180 patients with WHO grade I tumors and was significantly higher in two of nine patients with WHO grade II tumors. A volume reduction of more than 50% was observed in 26 patients (14%). Preexisting cranial nerve symptoms resolved completely in 28% of the patients. Clinically significant treatment-induced toxicity was seen in 1.6% of the patients. No treatment-related deaths occurred. CONCLUSION: The results of this study demonstrate that fractionated stereotactic radiotherapy is safe and effective in the therapy of subtotally resected or unresectable meningiomas. The overall morbidity and incidence subacute and late side effects of this conformal radiotherapy approach were low. PMID- 11481363 TI - Randomized clinical trials in oncology: understanding and attitudes predict willingness to participate. AB - PURPOSE: To explore the association at different time points in the trajectory of breast cancer care, between anxiety, knowledge, and attitudes, on women's willingness to participate in randomized clinical trials. MATERIALS AND METHODS: A cross-sectional survey was undertaken among women attending a breast clinic for screening mammography or diagnostic assessment plus women with newly diagnosed breast cancer to assess attitudes toward and willingness to participate in randomized clinical trials of breast cancer treatment. RESULTS: Five hundred forty-five women completed questionnaires assessing knowledge of and attitudes toward randomized clinical trials. The mean age of respondents was 48.9 years (SD, 11.3 years). Thirty-three percent of women would consider participating in a clinical trial if they had breast cancer. Women with breast cancer (31%) were significantly more likely to decline to participate than women attending for screening mammography (15%) or diagnostic assessment (15%, P =.0002). Women who might consider participating in a randomized clinical trial were more knowledgeable about randomized trials (mean difference, 0.7; 95% confidence interval [CI], 0.2 to 1.2; P =.003). In a multivariate analysis, women who would consider participating in a randomized trial were younger (odds ratio [OR], 0.96; 95% CI, 0.93 to 0.99), more likely to want an active role in decision-making (OR, 3.2; 95% CI, 1.3 to 7.6), and reported a greater impact from the positive aspects of clinical trials (OR, 2.2; 95% CI, 1.3 to 3.8) and less impact from the negative aspects of clinical trials (OR, 2.2; 95% CI, 1.3 to 3.2). CONCLUSION: These findings suggest that women who have a better understanding of issues about clinical trials have more favorable attitudes toward randomized trials and are more willing to consider participation in a clinical trial. PMID- 11481364 TI - Biology of osteoclast activation in cancer. AB - Bone is a frequent site of cancer metastasis. Bone metastases can result in bone destruction or new bone formation. Bone destruction is mediated by factors produced or induced by tumor cells that stimulate formation and activation of osteoclasts, the normal bone-resorbing cells. Local bone destruction also occurs in patients with osteoblastic metastases and may precede or occur simultaneously with increased bone formation. Several factors, including interleukin (IL)-1, IL 6, receptor activator of NF-kappaB (RANK) ligand, parathyroid hormone-related protein (PTHrP), and macrophage inflammatory protein-1-alpha (MIP-1alpha), have been implicated as factors that enhance osteoclast formation and bone destruction in patients with neoplasia. PTHrP seems to be the major factor produced by breast cancer cells that induces osteoclast formation through upregulation of RANK ligand. Enhanced RANK ligand expression also plays an important role in bone destruction in patients with myeloma. RANK ligand can act to enhance the effects of other factors produced by myeloma cells or in response to myeloma cells, such as MIP-1alpha and/or IL-6, to induce osteoclast formation. Understanding of the molecular mechanisms responsible for osteoclast activation in osteolytic metastases should lead to development of novel therapeutic approaches for this highly morbid and potentially fatal complication of cancer. PMID- 11481365 TI - Two unusual sites of colon cancer metastases and a rare thyroid lymphoma. Case 1. Metastatic colon cancer to a multinodular goiter. PMID- 11481366 TI - Two unusual sites of colon cancer metastases and a rare thyroid lymphoma. Case 2. Chemotherapy-responsive right artial metastasis from colon carcinoma. PMID- 11481367 TI - Two unusual sites of colon cancer metastases and a rare thyroid lymphoma. Case 3. Primary T-cell lymphoma of the thyroid arising in a background of Hashimoto's thyroiditis. PMID- 11481368 TI - Life after breast cancer. PMID- 11481369 TI - Is there an association between total-body irradiation and secondary acute myelogenous leukemia/myelodysplastic syndrome in patients with relapsed/refractory Hodgkin's disease treated with autologous stem-cell transplantation? PMID- 11481370 TI - Patients' needs at the end of life. PMID- 11481371 TI - Adjuvant chemotherapy for ependymoma: is it necessary for all children under five years of age? PMID- 11481372 TI - Is immunohistochemical analysis an appropriate diagnostic technique for bone marrow micrometastases? PMID- 11481373 TI - Scientific fraud and international research. PMID- 11481374 TI - Understanding mechanisms of breast cancer prevention. PMID- 11481375 TI - Are patterns of HER-2/neu amplification and expression among primary tumors and regional metastases indicative of those in distant metastases and predictive of Herceptin response? PMID- 11481376 TI - Ovarian tumors of low malignant potential: Can molecular biology solve this enigma? PMID- 11481377 TI - Creation of Center for Cancer Research marks cultural shift. PMID- 11481378 TI - Comparing microarray data: What technology is needed? PMID- 11481379 TI - Female cancer patients at higher risk of marital problems. PMID- 11481380 TI - IOM calls for improvements in palliative care. PMID- 11481381 TI - Government-industry collaborations explored for alternative medicine. PMID- 11481382 TI - Stat bite: Cancer survival rates for children under age 15 years in Great Britain. PMID- 11481383 TI - Enrollment begins for largest-ever prostate cancer prevention trial. PMID- 11481384 TI - Placental characteristics and reduced risk of maternal breast cancer. AB - BACKGROUND: Women who have preeclampsia during pregnancy are at reduced risk of subsequent breast cancer. We examined whether other markers of reduced placental size or function, including increased blood pressure during pregnancy, predict a reduction in maternal breast cancer. METHODS: The Child Health and Development Studies is a 40-year follow-up of pregnant women enrolled in the Kaiser Permanente health plan between 1959 and 1967. We identified 3804 white women for whom data were available on placental examinations and other study variables. As of 1997, 146 women had developed invasive breast cancer. Proportional hazards models were used to estimate associations of breast cancer with markers of placental function. All statistical tests were two-sided. RESULTS: A blood pressure increase between the second and third trimesters exhibited a linear relationship with breast cancer rate, with the highest quartile showing a 51% reduction (95% confidence interval [CI] = 20% to 70%) that was not explained by preeclampsia. Smaller placental diameter was independently associated with a reduced breast cancer rate; the association increased with age at first pregnancy (P =.008). Maternal floor infarction of the placenta was associated with a 60% reduction in breast cancer rate (95% CI = 12% to 82%). In combination, placental risk factors were associated with a reduction in the breast cancer rate of as high as 94% (95% CI = 80% to 98%). CONCLUSIONS: Smaller placentas, maternal floor infarction of the placenta, and increasing blood pressure during pregnancy were associated with reduced maternal breast cancer. In the case of smaller placental diameter, the larger reduction observed with older age at first pregnancy suggests a process in which promotion of an existing lesion is blocked. Elucidating the mechanisms for these associations could provide clues to breast cancer prevention and treatment. PMID- 11481385 TI - Patterns of her-2/neu amplification and overexpression in primary and metastatic breast cancer. AB - BACKGROUND: Only 25% of patients with HER-2/neu-positive metastatic breast tumors respond favorably to trastuzamab (Herceptin) treatment. We hypothesized that a high failure rate of patients on trastuzamab could result if some of the metastases were HER-2 negative and these metastases ultimately determine the course of the disease. METHODS: We used tissue microarrays (TMAs) containing four samples each from 196 lymph node-negative primary tumors, 196 lymph node-positive primary tumors, and three different lymph node metastases from each lymph node positive tumor to estimate HER-2 gene amplification by fluorescence in situ hybridization (FISH) and Her-2 protein overexpression by immunohistochemistry (IHC). RESULTS: FISH and IHC analyses gave the same result with respect to HER-2 status for 93.7% of the tissues contained in the TMAs. Tissue samples were, therefore, considered to be HER-2 positive if they were positive for either HER-2 DNA amplification or Her-2 protein expression and HER-2 negative if both FISH and IHC gave a negative result. The HER-2 status of lymph node-positive primary tumors was maintained in the majority of their metastases. For HER-2-positive primary tumors, 77% (95% confidence interval [CI] = 59% to 90%) had entirely HER 2-positive metastases, 6.5% (95% CI = 8% to 21%) had entirely HER-2-negative metastases, and 16.3% (95% CI = 5% to 34%) had a mixture of HER-2-positive and HER-2-negative metastases. For HER-2-negative primary tumors, 95% (95% CI = 88% to 98%) had metastases that were entirely negative for HER-2. CONCLUSIONS: Our data suggest that differences in HER-2 expression between primary tumors and their lymph node metastases cannot explain the high fraction of nonresponders to trastuzamab therapy. PMID- 11481386 TI - Molecular evidence for the independent origin of extra-ovarian papillary serous tumors of low malignant potential. AB - BACKGROUND: Molecular data suggest that peritoneal tumors in women with advanced stage ovarian papillary serous adenocarcinoma are monoclonal in origin. Whether the same is true for ovarian tumors of low malignant potential is not known. We compared peritoneal and ovarian tumors from women with advanced-stage ovarian papillary serous tumors of low malignant potential to determine whether the peritoneal tumors arose from the same clone as the ovarian tumors. METHODS: We studied the clonality of 73 peritoneal and ovarian tumors from 18 women with advanced-stage ovarian papillary serous tumors of low malignant potential. Formalin-fixed, paraffin-embedded tumors and representative normal tissues were sectioned and stained with hematoxylin-eosin, representative sections from separate tumors were manually microdissected, genomic DNA was extracted from the microdissected tumors, and the polymerase chain reaction was used to amplify a CAG polymorphic site in the human androgen receptor locus on the X chromosome to determine the inactivation pattern of the X chromosome and the clonality of the tumors. RESULTS: The pattern of X-chromosome inactivation could be determined from the tumors of 13 of 18 patients. Of the 13 patients, seven (54%) had nonrandom inactivation of the X chromosome, and six of the seven had different inactivation patterns in the peritoneal and ovarian tumors. Three of these patients also had different patterns of nonrandom X-chromosome inactivation in tumors from each ovary. The remaining six patients had random patterns of X chromosome inactivation in the peritoneal and ovarian tumors. CONCLUSIONS: Our data suggest that peritoneal and ovarian tumors of low malignant potential arise independently. PMID- 11481387 TI - Pathologic features of prostate cancer found at population-based screening with a four-year interval. AB - BACKGROUND: The currently recommended frequency for prostate-specific antigen (PSA) screening tests for prostate cancer is 1 year, but the optimal screening interval is not known. Our goal was to determine if a longer interval would compromise the detection of curable prostate cancer. METHODS: A cohort of 4491 men aged 55-75 years, all participants in the Rotterdam section of the European Randomized Study of (population-based) Screening for Prostate Cancer, were invited to participate in an initial PSA screening. Men who received that screening were invited for a second screen 4 years later. Pathology findings from needle biopsy cores were compared for men in both rounds. Statistical tests were two-sided. RESULTS: A total of 4133 men were screened in the first round (the prevalence screen), and 2385 were screened in the second round. The median amount of cancer in needle biopsy sets was 7.0 mm (95% confidence interval [CI] = 5.4 mm to 8.6 mm) in the first round and 4.1 mm (95% CI = 2.6 mm to 5.6 mm) in the second round (P =.001). Thirty-six percent of the adenocarcinomas detected in the first round but only 16% of those detected in the second round had a Gleason score of 7 or higher (mean difference = 20% [95% CI = 10% to 30%]; P<.001). Whereas 25% of the adenocarcinomas detected in the first round had adverse prognostic features, only 6% of those detected in the second round did (mean difference = 19% [95% CI = 11% to 26%]; P<.001). Baseline PSA values were predictive for the amount of tumor in biopsies in men with cancer in the first round but not for that in the second round. CONCLUSION: Most large prostate cancers with high serum PSA levels were effectively detected in a prevalence screen. In this population, a screening interval of 4 years appears to be short enough to constrain the development of large tumors, although it is inconclusive whether this will result in a survival benefit. PMID- 11481388 TI - RBP1L1, a retinoblastoma-binding protein-related gene encoding an antigenic epitope abundantly expressed in human carcinomas and normal testis. AB - BACKGROUND: Antibodies isolated from cancer patients have been used to identify genes encoding tumor-associated antigen epitopes relevant to immune responses in cancer patients. In this report, we used an immunoglobulin G (IgG) purified from serum of a patient with breast cancer to identify its corresponding epitope, gene, and protein-retinoblastoma-binding protein-1-like protein-1 (RBP1L1)-and determined whether it is a potential molecular marker for various cancers. METHODS: IgG purified from the serum of a patient with breast cancer was used to screen an MCF-7 breast cancer cell complementary DNA (cDNA) expression library for immunoreactive clones. The cDNAs identified were cloned and sequenced. Immunoreactivity of specific amino acids in the epitope was determined by western blot analysis and enzyme-linked immunosorbent assay. The cellular location of the antigen was determined by immunoperoxidase staining with purified RBP1L1-specific IgG. Gene expression in various human carcinomas and normal tissues was examined by northern blot analysis and the reverse transcription-polymerase chain reaction. RESULTS: Our purified IgG recognized just one epitope on RBP1L1. The complete 5802-base-pair RBP1L1 cDNA encodes a 1226-amino acid protein containing the antigenic epitope IKPSLGSKK. The derived protein sequence of RBP1L1 shares 74% and 37% amino acid identity, respectively, with a partial sequence of the retinoblastoma-binding protein and the complete sequence of retinoblastoma binding protein-1. The RBP1L1 epitope was localized to the cytoplasm of MCF-7 cells but was not detected in peripheral blood mononuclear cells. High expression of RBP1L1 messenger RNA was found in human breast, lung, colon, pancreatic, and ovarian cancers and in normal testis, but expression was limited in other normal tissues. CONCLUSIONS: RBP1L1 appears to be a molecular marker associated with a broad range of human malignancies. PMID- 11481389 TI - NAD(P)H:quinone oxidoreductase 1 deficiency and increased susceptibility to 7,12 dimethylbenz[a]-anthracene-induced carcinogenesis in mouse skin. AB - BACKGROUND: The phase II enzyme NAD(P)H :quinone oxidoreductase 1 (NQO1) catalyzes quinone detoxification, protecting cells from redox cycling, oxidative stress, mutagenicity, and cytotoxicity induced by quinones and its precursors. We have used NQO1(-/-) C57BL/6 mice to show that NQO1 protects them from skin cancer induced by the polycyclic aromatic hydrocarbon benzo[a]pyrene. Herein, we used NQO1(-/-) mice to investigate whether NQO1 also protects them against 7,12 dimethylbenz[a]anthracene (DMBA), where methyl substituents diminish primary quinone formation. METHODS: Dorsal skin of NQO1(-/-) or wild-type C57BL/6 mice was shaved. When tested as a complete carcinogen, DMBA (500 or 750 microg in 100 microL of acetone) alone was applied to the shaved area. When tested as a tumor initiator, DMBA (200 or 400 nmol in 100 microL of acetone) was applied to the shaved area; 1 week later, twice-weekly applications of phorbol 12-myristate 13 acetate (PMA)-10 microg dissolved in 200 microL of acetone-to the same area began and were continued for 20 weeks. Tumor development was monitored in all mice (12 15 per group). All statistical tests were two-sided. RESULTS: When DMBA (750 microg) was tested as a complete carcinogen, about 50% of the DMBA-treated NQO1( /-) mice but no DMBA-treated wild-type mouse developed skin tumors. When DMBA (both concentrations) was used as a tumor initiator, NQO1(-/-) mice developed larger tumors at a greater frequency than their wild-type littermates. Twenty three weeks after the first PMA treatment in the tumor initiator test, all 30 NQO1(-/-) mice given 400 nmol of DMBA had developed skin tumors, compared with 33% (10 of 30) of treated wild-type mice (P<.001). CONCLUSIONS: NQO1(-/-) mice are more susceptible to DMBA-induced skin cancer than are their wild-type littermates, suggesting that NQO1 may protect cells from DMBA carcinogenesis. PMID- 11481390 TI - cis-Activation of the human telomerase gene (hTERT) by the hepatitis B virus genome. PMID- 11481391 TI - Re: Association of African-American ethnic background with survival in men with metastatic prostate cancer. PMID- 11481392 TI - Re: Levels of hypoxia-inducible factor-1alpha during breast carcinogenesis. PMID- 11481396 TI - Insulin stimulation of hepatic triacylglycerol secretion and the etiology of insulin resistance. AB - The recent observations that insulin can either stimulate or inhibit triacylglycerol secretion by the liver, depending on prior metabolic (possibly insulinemic) state, have rationalized the many apparently contradictory observations, obtained over the past three decades, on the effects of the hormone on this aspect of hepatic metabolism. Extrapolation to the situation in vivo suggests that frequent stimulation of insulin secretion may result in a chronic stimulation of VLDL secretion, and increased delivery of acyl moieties to muscle, where they induce insulin resistance if provided in excess of the oxidative needs (mostly due to exercise) of the tissue. High fructose/sucrose diets, which also stimulate hepatic VLDL secretion, will have the same effect, especially if consumed frequently during the diurnal cycle. Due to the quantitative importance of muscle as a site for insulin-sensitive glucose metabolism, these effects may initiate the metabolic vicious cycle that results in the development of the metabolic syndrome, well in advance of overt obesity or the diagnosis of type-2 diabetes. PMID- 11481397 TI - Please pass the chips: genomic insights into obesity and diabetes. AB - Type 2 diabetes mellitus is an increasingly common disorder of carbohydrate and lipid metabolism. Approximately 16 million individuals in the United States have diabetes, and 800,000 new cases are identified each year. Two important characteristics of this disease are insulin resistance, the failure of peripheral tissues, including liver, muscle, and adipose tissue, to respond to physiologic doses of insulin, and failure of pancreatic beta-cells to properly secrete insulin in response to elevated blood glucose levels. Obesity is a significant risk factor for the development of type 2 diabetes mellitus. Recent observations of extremely lean, lipoatrophic models have revealed a similar predisposition to developing diabetes. Although it may seem paradoxical that both increased adiposity and severely reduced fat mass cause diabetes, a common pathophysiologic process in fat may be responsible for the predisposition to develop hyperglycemia in both conditions. This review will focus on the important role of adipose tissue dysfunction in the pathogenesis of diabetes, and on insights gained through the application of microarray technology to analyze adipocyte gene expression. PMID- 11481398 TI - Pomegranate juice supplementation to atherosclerotic mice reduces macrophage lipid peroxidation, cellular cholesterol accumulation and development of atherosclerosis. AB - Inhibition of lipid peroxidation contributes to the attenuation of macrophage cholesterol accumulation, foam-cell formation and atherosclerosis. Evidence suggests that nutritional antioxidants such as pomegranate juice (PJ) can contribute to the reduction of oxidative stress and atherogenesis. The goals of the present study were to determine whether such beneficial effects of PJ exist when supplemented to apolipoprotein E-deficient (E(0)) mice with advanced atherosclerosis and to analyze the antiatherosclerotic activity of a tannin fraction isolated from PJ. Mice (4-mo-old) were supplemented with PJ in their drinking water for 2 mo and compared with age-matched placebo-treated mice, as well as to young (4-mo-old) control mice, for their mouse peritoneal macrophage (MPM) oxidative state, cholesterol flux and mice atherosclerotic lesion size. PJ supplementation reduced each of the proatherogenic variables determined in the present study compared with age-matched placebo-treated mice. It significantly induced serum paraoxonase activity and reduced MPM lipid peroxide content compared with placebo-treated mice and control mice. PJ administration to E(0) mice significantly reduced the oxidized (Ox)-LDL MPM uptake by 31% and MPM cholesterol esterification and increased macrophage cholesterol efflux by 39% compared with age-matched, placebo-treated mice. PJ consumption reduced macrophage Ox-LDL uptake and cholesterol esterification to levels lower than those in 4-mo-old, unsupplemented controls. PJ supplementation to E(0) mice with advanced atherosclerosis reduced the lesion size by 17% compared with placebo treated mice. In a separate study, supplementation of young (2-mo-old) E(0) mice for 2 mo with a tannin fraction isolated from PJ reduced their atherosclerotic lesion size, paralleled by reduced plasma lipid peroxidation and decreased Ox-LDL MPM uptake. PJ supplementation to mice with advanced atherosclerosis reduced their macrophage oxidative stress, their macrophage cholesterol flux and even attenuated the development of atherosclerosis. Moreover, a tannin-fraction isolated from PJ had a significant antiatherosclerotic activity. PMID- 11481399 TI - Curcumin and especially tetrahydrocurcumin ameliorate oxidative stress-induced renal injury in mice. AB - Protective effects of curcumin (U1), one of the major yellow pigments in turmeric and its derivative, tetrahydrocurcumin (THU1), against ferric nitrilotriacetate (Fe-NTA)-induced oxidative renal damage were studied in male ddY mice. Single Fe NTA treatment (5 mg Fe/kg body intraperitoneally) transiently causes oxidative stress, as shown by the accumulation of lipid peroxidation products and 8-hydroxy 2'-deoxyguanosine in the kidney. Mice were fed with a diet containing 0.5 g/100 g U1 or THU1 for 4 wk. THU1 significantly inhibited 2-thiobarbituric acid reactive substances and 4-hydroxy-2-nonenal-modified proteins and 8-hydroxy-2' deoxyguanosine formation in the kidney; U1 inhibited only 4-hydroxy-2-nonenal modified protein formation. To elucidate the mechanisms of protection by U1 and THU1, the pharmacokinetics and radical-scavenging capacities of U1 and THU1 were investigated by HPLC and electron spin resonance spin trapping with 5,5-dimethyl 1-pyrroline-N-oxide, respectively. Induction of antioxidant enzymes was also investigated. The amounts of THU1 and its conjugates (as sulfates and glucuronides) in the liver and serum were larger in the THU1 group than in the U1 group. The amounts of U1 and its conjugates were small even in the U1 group. These results suggest that THU1 is more easily absorbed from the gastrointestinal tract than U1. Furthermore, THU1 induced antioxidant enzymes, such as glutathione peroxidase, glutathione S-transferase and NADPH: quinone reductase, as well as or better than U1 and scavenged Fe-NTA-induced free radicals in vitro better than U1. These results suggest that U1 is converted to THU1 in vivo and that THU1 is a more promising chemopreventive agent. PMID- 11481400 TI - Serum carotenoid depletion follows first-order kinetics in healthy adult women fed naturally low carotenoid diets. AB - Dietary intakes of carotenoids are highly variable in human populations as are serum carotenoid concentrations. However, there are few controlled data relating carotenoid intake to concentration. Most of the data that are available are from measurements of the absorption and decay of large pharmacologic doses of carotenoids, and are therefore of unknown physiologic relevance. Our objective was to determine the half-life (t(1/2)) of the most abundant carotenoids in blood serum from healthy adult women living under controlled conditions. As part of two carotenoid isotopic studies, we measured serum concentrations of beta-carotene, alpha-carotene, lutein, zeaxanthin, beta-cryptoxanthin and lycopene in 19 healthy young adult women that were fed controlled low carotenoid diets for approximately 10 wk. All other nutrients (vitamins A, E and C) were provided at 100-150% of the 1989 U.S. recommended dietary allowance levels. Exercise and activities were controlled throughout the studies to simulate usual activity patterns. Carotenoid concentrations were measured by reversed-phase HPLC. Serum carotenoid concentration decreases during depletion followed first-order kinetics. The half lives determined in decreasing order were as follows: lutein (76 d) > alpha carotene (45 d) = beta-cryptoxanthin (39 d) = zeaxanthin (38 d) = beta-carotene (37 d) > lycopene (26 d). Half-lives were unrelated to physical or demographic characteristics such as body mass, body fat, racial background or age in these relatively homogeneous groups. Carotenoids decreased by similar first-order mechanisms, although the rates differed for individual carotenoids. PMID- 11481401 TI - Gastric digestion of bovine lactoferrin in vivo in adults. AB - Lactoferrin (LF), an iron-binding glycoprotein present in milk and other endocrine and exocrine secretions, may exert a number of physiologic effects in the intestines. To study the effects of oral LF supplementation in vivo in the gastrointestinal tract, information about the gastric survival of LF in vivo is important. We tested 12 healthy volunteers (age 21 +/- 0.3 y) on 3 separate d according to a randomized, cross-over design. A test drink containing 4.5 g of bovine LF (20% iron-saturated LF; apoLF) in the presence of a gastric pH buffer (0.1 mol/L sodium citrate/citric acid; apoLFbuf), apoLF without the buffer (apoLF) or iron-saturated LF (holoLF) was administered into the stomach using nasogastric intubation. Gastric emptying rate, determined by a marker dilution technique, did not differ among any of these drinks. Gastric survival of LF, analyzed by gel permeation chromatography under denaturing conditions, was 64%, 62% and 79% after consumption of the apoLFbuf, apoLF and holoLF test drinks, respectively. Addition of the gastric pH buffer initially lowered intragastric pH because of its hydroxide buffering effect. However, it did not elevate intragastric pH over a prolonged period and thereby inhibit intragastric LF breakdown. We conclude that after oral administration, substantial amounts of apoLF and holoLF survive gastric transit. PMID- 11481402 TI - Long-term ingestion of a fermented soybean-derived Touchi-extract with alpha glucosidase inhibitory activity is safe and effective in humans with borderline and mild type-2 diabetes. AB - Water-extracted Touchi, a traditional Chinese food, exerts a strong inhibitory activity against rat intestinal alpha-glucosidase in foodstuffs, and Touchi extract (TE) has been shown to have an antihyperglycemic effect in rats and humans after a single oral administration. In the present complementary study, the effects of powdered Houji-tea with or without (placebo) TE, a formula designed to enhance good compliance, were monitored in a 3-mo double-blind randomized group comparison study with placebo controls in humans with borderline and mild type-2 diabetes (n = 36). All subjects ingested Houji-tea with or without 0.3 g of TE before each of three meals per day for 3 mo. In the TE group, initial fasting blood glucose (6.9 +/- 0.1 mmol/L) and glycated hemoglobin (HbA(1c); 6.1 +/- 0.1%) levels gradually decreased; fasting blood glucose decreased significantly after 3 mo (6.4 +/- 0.3 mmol/L; P < 0.05) as did HbA(1c) (5.6 +/- 0.2%; P < 0.01) levels at 2 mo postingestion of TE and thereafter. In contrast, fasting blood glucose and HbA(1c) levels did not change in the placebo group. In this study, other biochemical variables were not affected in any of the subjects, and no one complained of any side effects or abdominal distension. Moreover, there was no deterioration as assessed by fasting blood glucose and HbA(1c) levels after withdrawal of TE ingestion. Thus, the alpha-glucosidase inhibitory TE demonstrated an antihyperglycemic effect and may prove useful for improving glycemic control in subjects suffering from borderline and type-2 diabetes mellitus. PMID- 11481403 TI - Comparison of the intestinal absorption of quercetin, phloretin and their glucosides in rats. AB - Absorption and metabolism of quercetin and isoquercitrin (quercetin 3-O-glucose) were investigated in rats after in situ perfusion of jejunum plus ileum (15 nmol/min) for 30 min and compared with those of phloretin and phloridzin (phloretin 2'-O-glucose). After perfusion of the glucosides, the corresponding aglycone forms and conjugated derivatives appeared in the lumen. The conjugated metabolites were similar to those recovered after intestinal perfusion of the aglycone forms. Regardless of the aglycone or glucoside perfused, only conjugated forms were present in the mesenteric vein blood draining the perfused segment showing the importance of intestinal conjugation. The hydrolysis of glucosides was a prerequisite step before their conjugation by intestinal enzymes and their transport towards the mucosal and serosal sides. In contrast to phloridzin, lactase phloridzin hydrolase activity did not seem to be an essential pathway for isoquercitrin hydrolysis. The 3-O-glucosylation of quercetin improved the net absorption of the aglycone (P < 0.05), whereas phloretin absorption decreased when present as 2'-O-glucoside (P < 0.05). Whatever the perfused compound, the efficiency of the absorption seemed to be linked to the intestinal conjugation process and to the luminal secretion of metabolites. PMID- 11481404 TI - Palmitic and stearic acids similarly affect plasma lipoprotein metabolism in cynomolgus monkeys fed diets with adequate levels of linoleic acid. AB - This study was designed to evaluate whether the exchange of specific saturated fatty acids [SFA; palmitic acid (16:0) for stearic acid (18:0)] would differentially affect plasma lipids and lipoproteins, when diets contained the currently recommended levels of total SFA, monounsaturated fatty acids and polyunsaturated fatty acids (PUFA). Ten male cynomolgus monkeys were fed one of two purified diets (using a cross-over design) enriched either in 16:0 (palmitic acid diet) or 18:0 (stearic acid diet). Both diets provided 30% of energy as fat (SFA/monounsaturated fatty acid/PUFA: 1/1/1). The palmitic acid and stearic acid diets were based on palm oil or cocoa butter (59% and 50% of the total fat, respectively). By adding different amounts of sunflower, safflower and olive oils, an effective exchange of 16:0 for 18:0 of approximately 5% of energy was achieved with all other fatty acids being held constant. Monkeys were rotated through two 10-wk feeding periods, during which time plasma lipids and in vivo lipoprotein metabolism (following the simultaneous injection of (131)I-LDL and (125)I- HDL were evaluated). Plasma triacyglycerol (0.40 +/- 0.03 vs. 0.37 +/- 0.03 mmol/L), plasma total cholesterol (3.59 +/- 0.18 vs. 3.39 +/- 0.23 mmol/L), HDL cholesterol (1.60 +/- 0.16 vs 1.53 +/- 0.16 mmol/L) and non-HDL cholesterol (2.02 +/- 0.26 vs. 1.86 +/- 0.23 mmol/L) concentrations did not differ when monkeys consumed the palmitic acid and stearic acid diets, respectively. Plasma lipoprotein compositional analyses revealed a higher cholesteryl ester content in the VLDL fraction isolated after consumption of the stearic acid diet (P < 0.10), as well as a larger VLDL particle diameter (16.3 +/- 1.7 nm vs. 13.8 +/- 3.6 nm; P < 0.05). Kinetic analyses revealed no significant differences in LDL or HDL transport parameters. These data suggest that when incorporated into diets following current guidelines, containing adequate PUFA, an exchange of 16:0 for 18:0, representing approximately 11 g/(d.10.46 mJ) [ approximately 11 g/(d.2500 kcal)] does not affect the plasma lipid profile and has minor effects on lipoprotein composition. Whether a similar effect would occur in humans under comparable dietary conditions remains to be established. PMID- 11481405 TI - Functional fragments of ingested lactoferrin are resistant to proteolytic degradation in the gastrointestinal tract of adult rats. AB - Pharmaceutical and food-related applications of lactoferrin, an 80-kDa iron binding glycoprotein found predominantly in milk, have attracted interest lately, but the process of digestion of lactoferrin has been poorly characterized. The digestive fate of bovine lactoferrin in adult rats after oral administration of a single dose and after dietary supplementation was studied by (125)I-labeling and by surface-enhanced laser desorption/ionization (SELDI) affinity mass spectrometry. The latter method was designed to detect multiple forms of degraded lactoferrin as simple molecular ion peaks corresponding to one of the core regions of lactoferrin, namely, the lactoferricin region (Phe17-Ala42). Radioactive fragments with molecular masses of 42, 36, 33 and 29 kDa were observed at 20, 60 and 180 min postingestion in the contents of the lower small intestine. Rats were given free access to milk enriched with lactoferrin at 482 micromol/L (40 mg/mL). The concentrations of lactoferrin fragments in the contents of the stomach, small intestine and lower small intestine as determined by SELDI affinity mass spectrometry were approximately 200, 20 and 1 micromol/L, respectively. These data indicate that functional fragments of LF such as fragments containing glycosaminoglycan-binding site(s), as well as large fragments with a mass >20 kDa, indeed survive proteolytic degradation in the small intestine of adult rats. PMID- 11481406 TI - Both L- and D-lactate contribute to metabolic acidosis in diarrheic calves. AB - Diarrhea in neonates is often complicated by metabolic acidosis. We used blood gas analysis and HPLC to determine whether bacterial fermentation might contribute to acidosis in diarrheic calves. Diarrheic calves (n = 21) had significantly lower pH, PCO(2), HCO(3)(-) and a higher anion gap than healthy calves (n = 21). Serum concentrations (mean +/- SD, mmol/L) of DL-, L- and D lactate were also significantly higher in diarrheic (8.9 +/- 5.1, 4.1 +/- 3.4 and 5.2 +/- 5.7) than in healthy calves (1.7 +/- 1.2, 2.0 +/- 1.1 and too low to quantify). D- and L-lactate accounted for 64% anion gap increase in diarrheic calves. Fecal D- and L-lactate concentrations were also significantly higher in diarrheic calves (9.4 +/- 3.0 and 11.9 +/- 2.7 mmol/L) than healthy calves (1.1 +/- 0.1 and 1.6 +/- 0.1 mmol/L). The elevated concentrations of serum and fecal D lactate suggest gut bacterial fermentation contributes to the development of acidosis in diarrhea. PMID- 11481407 TI - Dietary zinc deficiency and repletion modulate metallothionein immunolocalization and concentration in small intestine and liver of rats. AB - Metallothionein (MT) functions in zinc (Zn) homeostasis and dietary Zn affects tissue MT concentration. The objective of this study was to investigate the effects of dietary Zn deficiency and 24-h Zn repletion on MT immunolocalization and concentration in the small intestine and liver of growing rats. Three-week old rats fed Zn-deficient diet (< 1 mg Zn/kg) for 16 d had no MT staining in either small intestine or liver. After 24-h Zn repletion with control diet (30 mg Zn/kg), strong MT staining was observed in intestinal Paneth cells and surface epithelial cells in the proliferative regions of villi. Pair-fed control rats had strong MT staining in liver that was localized around central veins. After 24-h energy repletion, the hepatic MT staining diminished. Furthermore, Zn-deficient rats had significantly reduced intestinal (57%) and hepatic (61%) MT concentrations but unaffected Zn concentrations compared with controls that consumed food ad libitum. Zn repletion for 24 h restored intestinal and hepatic MT concentrations and reduced hepatic Zn concentration. Pair-fed control rats had elevated MT concentration in liver that was normalized by energy repletion. There was a significant positive correlation between tissue Zn and MT concentrations in liver (r = 0.60, P = 0.0001), but not in small intestine. In summary, MT immunolocalization and concentration in rat small intestine and liver were responsive to changes in Zn status, supporting the role of MT in Zn metabolism. Cell-type-specific localization of MT in small intestine after dietary Zn manipulations indicates a function of Zn and MT in gut immunity and intestinal mucosal turnover, and the pattern of hepatic MT distribution with energy restriction may be linked to detoxification processes. PMID- 11481408 TI - Inflammatory responses to lipopolysaccharide are suppressed in 40% energy restricted mice. AB - To elucidate the suppressive effects of energy restriction on the inflammatory responses to lipopolysaccharide (LPS), mice were divided into a control group (fed 5.0 g diet/d; 71 kJ/d) and a 40% energy-restricted group (fed 3.0 g diet/d; 43 kJ/d) at 8-wk of age. Four weeks later, 25 microg of LPS was intraperitoneally injected. After the LPS injection, interleukin-1beta, interleukin-6 and tumor necrosis factor-alpha were elevated in serums in the 40% energy-restricted mice and in the controls, but the extent of the elevation was significantly lower in the restricted group. The LPS-induced expression of inducible nitric oxide synthase in the liver was significantly suppressed by the energy restriction. In addition, the LPS-induced elevations of serum aspartate and alanine aminotransferase activities, which are indexes of hepatic injury, were also significantly attenuated in the restricted group. Moreover, the extent of LPS induced alterations in hepatic structure was less in the restricted mice than in controls. Serum corticosterone level in the restricted mice was higher than that in the controls before LPS treatment (P < 0.05). Furthermore, after LPS injection, the significantly higher level of corticosterone was maintained in the restricted mice, although the LPS treatment significantly enhanced the level even in the control group. These results suggest that the extreme inflammatory responses to endotoxin are prevented in the 40% energy-restricted mice, and corticosterone participates in the preventive effects. PMID- 11481409 TI - Olive oil decreases both oxidative stress and the production of arachidonic acid metabolites by the prostaglandin G/H synthase pathway in rat macrophages. AB - Fish oil has a preventive role in cardiovascular and inflammatory diseases, but little is known about the effect of olive oil, which is widely consumed in Mediterranean regions. We examined the influence of dietary olive oil, corn oil and fish oil-rich diets on the production of superoxide anion (O2-) and nitric oxide (.NO) by resident macrophages stimulated by phorbol 12-myristate 13-acetate (PMA) and their effect on arachidonic acid release, prostaglandin G/H synthase-2 (PGHS-2) expression and the subsequent prostaglandin E(2) production. Resident peritoneal macrophages stimulated by PMA from rats fed with olive oil or corn oil had the same level of O2- production, but these levels were increased by the fish oil diet. Olive oil and the fish oil diets increased .NO and decreased arachidonic acid mobilization and the production of prostaglandin E(2). PGHS-2 expression, however, was not affected by diet. We conclude that although olive oil and fish oil reduce arachidonic acid mobilization and subsequent metabolism through the PGHS-2 pathway in PMA-stimulated macrophages, only olive oil offers an additional beneficial effect by increasing .NO/O2- production. PMID- 11481410 TI - Garlic supplementation increases testicular testosterone and decreases plasma corticosterone in rats fed a high protein diet. AB - The effects of garlic supplementation on protein metabolism were investigated by measuring testis testosterone and plasma corticosterone in rats fed diets with different protein levels. In Experiment 1, rats were fed experimental diets with different protein levels (40, 25 or 10 g/100 g casein) with or without 0.8 g/100 g garlic powder. After 28 d of feeding, testosterone contents in the testis were significantly higher and plasma corticosterone concentrations were significantly lower in rats fed 40 and 25% casein diets with garlic powder than in those fed the same diets without garlic powder. Urinary excretion of 17-ketosteroid (an index of testosterone), nitrogen balance and hepatic arginase activity were significantly higher in rats fed the 40% casein diet with garlic powder than in the 40% casein controls. In Experiment 2, the effect of diallyldisulfide (a major volatile sulfur-containing compound in garlic) on the secretion of luteinizing hormone (LH) from the pituitary gland, which regulates testosterone production in the testis, was investigated in anesthetized rats. Plasma LH concentration increased dose dependently after administration of diallyldisulfide (P < 0.01, r = 0.558). These results suggest that dietary supplementation with 0.8 g/100 g garlic alters hormones associated with protein anabolism by increasing testicular testosterone and decreasing plasma corticosterone in rats fed a high protein diet. PMID- 11481411 TI - Randomized structured triglycerides increase lymphatic absorption of tocopherol and retinol compared with the equivalent physical mixture in a rat model of fat malabsorption. AB - Previously we demonstrated that the digestion, absorption and lymphatic transport of lipid and key essential fatty acids (EFA) from randomly interesterified fish oil/medium-chain structured triglycerides (STG) were significantly higher than an equivalent physical mixture (PM) in a normal lymph fistula rat model and in a rat model of lipid malabsorption caused by ischemia/reperfusion (I/R) injury. The goals of this study were to further explore the potential absorptive benefits of STG by comparing the intestinal absorption and lymphatic transport of tocopherol and retinol when delivered gastrically with either STG or PM under normal conditions and after I/R injury to the small bowel. Food-deprived male Sprague Dawley rats were randomly assigned to two treatments (sham controls or I/R). Under halothane anesthesia, the superior mesenteric artery (SMA) was occluded for 20 min and then reperfused in I/R rats. The SMA was isolated but not occluded in control rats. In both groups, the mesenteric lymph duct was cannulated and a gastric tube was inserted. Each treatment group received 1 mL of the fish oil/MCT STG or PM (7 rats/group) along with (14)C-alpha-tocopherol and (3)H-retinol through the gastric tube followed by an infusion of PBS at 3 mL/h for 8 h. Lymph was collected hourly for 8 h. Under steady-state conditions, the amount of (14)C alpha-tocopherol and (3)H-retinol transported into lymph was significantly higher in the STG-fed rats compared with those fed PM in both control and I/R groups. In addition, control and I/R rats given STG had earlier steady-state outputs of (14)C-alpha-tocopherol and (3)H-retinol and maintained approximately 30% higher outputs in lymph throughout the 8-h lymph collection period compared with rats given the PM. We conclude that STG provides the opportunity to potentiate improved absorption of fat-soluble vitamins under normal and malabsorptive states. PMID- 11481412 TI - A glucagon-like peptide-1 receptor agonist and an antagonist modify macronutrient selection by rats. AB - The hypothesis that peripheral glucagon-like peptide-1 (GLP-1) is a regulator of both food intake and macronutrient selection in rats was tested by administration of its antagonist, exendin 9-39, and its agonist, exendin 4. The effect of exendin 9-39 given intraperitoneally (i.p.) on food intake was measured after carbohydrate, protein or fat preloads, and on choice between a protein-free, high carbohydrate (CHO) diet and a high protein, low carbohydrate (PRO) diet. The effect of exendin 4 on selection between the CHO and PRO diets was also investigated. Exendin 9-39 significantly enhanced food intake suppression occurring after glucose, but not after corn oil or albumin preloads. In diet selection studies, exendin 9-39 selectively decreased intake of only the CHO diet. In contrast, exendin 4 decreased intake of only the PRO diet. Thus, we suggest that peripheral GLP-1 plays a role in the regulation of macronutrient selection as well as food intake in rats. PMID- 11481413 TI - A longitudinal investigation of aggregate oral intake of copper. AB - Four-day composite solid food and beverage duplicate plates and 1-L samples of drinking water were collected from a stratified random sample of 80 individuals as part of the National Human Exposure Assessment Survey in Maryland. The media were obtained from each participant in up to six equally spaced sampling cycles over a year and analyzed for copper by inductively coupled plasma mass spectrometry. Copper concentrations (microg/kg) and consumption rates (kg/d) of solid food, beverage and drinking water were used to derive average daily aggregate oral intake of copper (microg/d). The mean aggregate copper intake of 263 measurements obtained from 68 people was 923.2 +/- 685.6 microg/d (mean +/- SD). Intake through solid food accounted for the majority of aggregate daily intake of copper contributing 87% on average. According to results from mixed model analysis of variance procedures, the mean log-transformed average daily copper intake in each medium except beverage exhibited significant (P < 0.05) variability among sampling cycles. Between-person variability accounted for 50% of the total variance in aggregate copper intake. As measured by the coefficient of variation, distributions of copper intake consisting of one observation per individual were more variable than the distribution consisting of the long-term average intake for each person. These results suggest that estimates of the fraction of a population at risk from chronic copper deficiency or excess copper intake can be overestimated if based upon short-term measures of copper intake. In addition, these results indicate that longitudinal information is required for accurate assessment of aggregate oral intake of copper for an individual. PMID- 11481414 TI - Fortification contributed greatly to vitamin and mineral intakes in the United States, 1989-1991. AB - The objective of this work was to quantify the contribution of fortification (defined here as adding nutrients beyond traditional enrichment standards) to dietary nutrient intakes in the United States. A list of fortified foods was developed that was relevant at the time of the analyses, and prefortification (naturally occurring) nutrients in the fortified foods were determined from industry-supplied data. Using dietary data from the 1989-1991 Continuing Survey of Food Intakes by Individuals (CSFII), intakes of nine nutrients were determined both as reported in the CSFII (i.e., postfortification) and also by using prefortification nutrient levels for the identified fortified foods. We report data for the total population age >/= 1 y based on respondents (n = 11,710) with 3 d of dietary data, as well as select age/gender subgroups. All data were weighted. Fortification substantially increased the intakes of all nutrients examined except calcium, in all age/gender groups but especially in children. In numerous cases, fortification was responsible for boosting median or 25th percentile intakes from below to above the RDA. The breakfast cereal category was responsible for nearly all the intake of nutrients from fortified foods, except vitamin C for which juice-type beverages made as great or a greater contribution. These data from 1989 to 1991 serve as a useful baseline with which to compare contributions of fortification as the practice expands. The large contribution of fortification even in 1989-1991 suggests that continued monitoring of fortification practices, using methods such as those presented here, is important. PMID- 11481415 TI - Serum concentrations of retinol, alpha-tocopherol and the carotenoids are influenced by diet, race and obesity in a sample of healthy adolescents. AB - An important part of understanding the functions of vitamin A, vitamin E and the carotenoids in nutritional status assessment, health promotion and disease prevention is knowledge of factors that influence their distribution in human tissues. Our objective was to examine serum concentrations of these nutrients and compounds in a sample of 285 healthy participants, 12-17 y old, from three U. S. cities. Pearson correlations between diet measured with a food frequency questionnaire and serum nutrient concentrations among these adolescents (adjusted for total serum cholesterol, age, sex, race and body mass index) were as follows: retinol, 0.23; alpha-tocopherol, 0.16; alpha-carotene, 0.31; beta-carotene, 0.15; beta-cryptoxanthin, 0.38; lycopene, 0.08; and lutein + zeaxanthin, 0.25. Multivariate linear regression modeled associations of demographic, dietary and physiologic variables with serum concentrations of these nutrients. African American participants had significantly lower concentrations of serum retinol (P < 0.001), alpha-tocopherol (P < 0.01) and alpha-carotene (P < 0.02), but higher concentrations of lutein + zeaxanthin (P = 0.001) compared with Caucasians. Obese participants had serum nutrient concentrations that were 2-10% (P < 0.05) lower than normal weight participants. Dietary intake was a significant predictor of all serum analytes (P < 0.01) except lycopene. These models explained 20% of the variability in serum retinol, 28% of the variability in serum alpha-tocopherol, and 14-24% of the variability in serum carotenoids. PMID- 11481416 TI - Inadequate nutrient intakes are common and are associated with low diet variety in rural, community-dwelling elderly. AB - Poor dietary habits and inadequate nutrient intakes are of concern in the elderly. The nutritional characteristics of those who survive to become the oldest are not well defined. Our goal was to describe dietary habits, nutrient intakes and nutritional risk of community-dwelling, rural Iowans, 79 y of age and older. Subjects were interviewed (n = 420) using a standardized format on one occasion in their homes and instructed to complete 3-d diet records (n = 261) after the in-home interview. Standardized interviews assessed demographic information, cognitive function and dietary habits (Nutrition Screening Initiative Checklist). Adequate nutrient intake was defined as consumption of the nutrient's estimated average requirement, 67% adequate intake or 67% recommended dietary allowance. Mean age was 85.2 y, 57% lived alone and 58% were widowed. Subjects completing 3-d diet records were younger, more cognitively intact and less likely to be at nutritional risk than subjects not completing diet records. The percentage of subjects with inadequate intakes of selected nutrients was 75% for folate, 83% for vitamin D and 63% for calcium. Eighty percent of subjects reported inadequate intakes of four or more nutrients. Diet variety was positively associated with the number of nutrients consumed at adequate intakes (r = 0.498), total energy (r = 0.522) and dietary fiber (r = 0.421). Our results suggest that rural, community-dwelling old have inadequate intakes of several nutrients. Recommendations to increase diet variety and consume a nutrient supplement may be necessary for elderly people to achieve adequate nutrient intakes. PMID- 11481417 TI - Downregulation of the cyclin D1/Cdk4 complex occurs during resveratrol-induced cell cycle arrest in colon cancer cell lines. AB - Resveratrol is a naturally occurring polyphenol with cancer chemopreventive properties. The objective of the current study was to investigate the effect of resveratrol on the human colonic adenocarcinoma cell line Caco-2. The compound inhibited cell growth and proliferation of Caco-2 cells in a dose-dependent manner (12.5-200 micromol/L) as assessed by crystal violet assay, [(3)H]thymidine and [(14)C]leucine incorporation. Furthermore, apoptosis was determined by measuring caspase-3 activity, which increased significantly after 24 and 48 h of treatment with 200 micromol/L resveratrol. Perturbed cell cycle progression from the S to G2 phase was observed for concentrations up to 50 micromol/L, whereas higher concentrations led to reversal of the S phase arrest. These effects were specific for resveratrol; they were not observed after incubation with the stilbene analogs stilbenemethanol and rhapontin. Levels of cyclin D1 and cyclin dependent kinase (cdk) 4 proteins were decreased, as revealed by immunoblotting. In addition, resveratrol enhanced the expression of cyclin E and cyclin A. The protein levels of cdk2, cdk6 and proliferating cell nuclear antigen were unaffected. Similar results were obtained for the colon carcinoma cell line HCT 116, indicating that cell cycle inhibition by resveratrol is independent of cyclooxygenase inhibition. The phosphorylation state of the retinoblastoma protein in Caco-2 cells was shifted from hyperphosphorylated to hypophosphorylated at 200 micromol/L, which may account for reversal of the S phase block at concentrations exceeding 50 micromol/L. These findings suggest that resveratrol exerts chemopreventive effects on colonic cancer cells by inhibition of the cell cycle. PMID- 11481418 TI - Vitamin B-6-supplemented diets compared with a low vitamin B-6 diet suppress azoxymethane-induced colon tumorigenesis in mice by reducing cell proliferation. AB - Male ICR mice were examined for the effect of vitamin B-6 [pyridoxine (PN) HCl] on azoxymethane-induced colon tumorigenesis. Mice were fed the diets containing 1, 7, 14 or 35 mg PN HCl/kg for 22 wk, and given a weekly injection of azoxymethane (5 mg/kg body) for the initial 10 wk. Compared with the 1 mg PN HCl/kg diet, 7, 14 and 35 mg PN HCl/kg diets significantly suppressed the incidence and number of colon tumors, colon cell proliferation and expressions of c-myc and c-fos proteins. For some variables, 14 and 35 mg PN HCl/kg diets were more effective than the 7 mg/kg diet. Supplemental vitamin B-6 had no influence on the number of colon apoptotic cells. The results suggest that elevating dietary vitamin B-6 suppresses colon tumorigenesis by reducing cell proliferation. PMID- 11481419 TI - A direct streptavidin-binding assay does not accurately quantitate biotin in human urine. AB - In human urine, the biotin concentration assayed directly using an avidin-binding assay (ABA) apparently overestimates "true" biotin concentration as measured by HPLC separation of biotin from biotin metabolites followed by ABA. Because biotin metabolites account for about half of biotin plus biotin metabolites in human urine, we speculate that the error might arise from biotin metabolites. We sought to test the following hypothesis: biotin measured by direct ABA routinely exceeds true biotin in urine due to biotin metabolites; however, if urinary biotin is quantitated using a streptavidin-binding assay (SABA) that does not detect biotin metabolites, results will agree with true biotin. An assay for biotin that uses europium coupled to streptavidin and time-resolved fluorescence was developed and validated. Urine samples were obtained from biotin-deficient, normal and biotin supplemented adults. In 133 urine samples from 26 subjects, biotin by direct ABA correlated positively and significantly with biotin measured after HPLC separation (P < 0.001; r = 0.78). However, biotin by direct ABA routinely exceeded true biotin. The magnitude of the overestimate correlated strongly with biotin metabolites; r = 0.80 and P < 0.0001. In 92 samples from nine subjects, biotin by direct SABA correlated positively and significantly with true biotin (P = 0.001; r = 0.73) but exceeded true biotin by more than analytical error in 62 of the 92 samples. The error did not correlate significantly with total biotin metabolites. In 62 samples analyzed by both assays, biotin by direct SABA correlated weakly (r = 0.69) but significantly (P < 0.0001) with biotin by direct ABA. These studies provide evidence that direct SABA does not accurately quantitate biotin. Although the errors from direct ABA arise primarily from metabolites, the errors from direct SABA cannot be attributed primarily to biotin metabolites. Whether these interfering substances are biotin metabolites or other unknown substances, the substances are likely separated from the biotin fraction by HPLC. PMID- 11481420 TI - Indirect calorimetry protocol development for measuring resting metabolic rate as a component of total energy expenditure in free-living postmenopausal women. AB - An objective measure of energy intake is needed in epidemiologic studies to evaluate random and systematic error associated with dietary self-report tools. Total energy expenditure in weight-stable humans is accepted as a measure of energy intake, but doubly labeled water remains cost prohibitive for large studies. Our purpose was to develop a practical indirect calorimetry (IC) protocol for estimating resting metabolic rate (RMR) in free-living, postmenopausal women. We conducted duplicate IC measures 1 wk apart using a canopy system on 102 women ages 50-79 y from the Seattle area. We compared RMR for 0-5, 5-10, 5-15, 5-20, 5-25, 5-30, and 0- to 30-min IC segments and segments meeting stability criteria. The mean RMR for the first 5 min was significantly higher than other time segments (P = 0.001). Correlation coefficients between duplicate measures were high (r = 0.90). Use of defined stability criteria produced RMR measures that were 10-30 kcal (42-126 kJ) higher than the 5- to 10 min RMR measures and 40-60% of subjects did not achieve these stability criteria. For protocols including IC to assess RMR as a component of total energy expenditure in free-living, postmenopausal women, a single 10-min canopy study, excluding the first 5 min of data, produces reliable results with minimal subject burden. PMID- 11481421 TI - Model for a third-party review of the evidence substantiating food and dietary supplement claims. PMID- 11481423 TI - Primary defects in the lens underlie complex anterior segment abnormalities of the Pax6 heterozygous eye. AB - We describe lens defects in heterozygous small eye mice, and autonomous deficiencies of Pax6(+/-) cells in the developing lens of Pax6(+/+) <--> Pax6(+/ ) chimeras. Two separate defects of the lens were identified by analyzing the distribution of heterozygous cells in chimeras: Pax6(+/-) cells are less readily incorporated into the lens placode than wild type, and those that are incorporated into the lens are not maintained efficiently in the proliferating lens epithelium. The lens of chimeric eyes is, therefore, predominantly wild type from embryonic day 16.5 onwards, whereas heterozygous cells contribute normally to all other eye tissues. Eye size and defects of the iris and cornea are corrected in fetal and adult chimeras with up to 80% mutant cells. Therefore, these aspects of the phenotype may be secondary consequences of primary defects in the lens, which has clinical relevance for the human aniridia (PAX6(+/-)) phenotype. PMID- 11481424 TI - DNA damage-inducible gene p33ING2 negatively regulates cell proliferation through acetylation of p53. AB - The p33ING1 protein is a regulator of cell cycle, senescence, and apoptosis. Three alternatively spliced transcripts of p33ING1 encode p47ING1a, p33ING1b, and p24ING1c. We cloned an additional ING family member, p33ING2/ING1L. Unlike p33ING1b, p33ING2 is induced by the DNA-damaging agents etoposide and neocarzinostatin. p33ING1b and p33ING2 negatively regulate cell growth and survival in a p53-dependent manner through induction of G(1)-phase cell-cycle arrest and apoptosis. p33ING2 strongly enhances the transcriptional transactivation activity of p53. Furthermore, p33ING2 expression increases the acetylation of p53 at Lys-382. Taken together, p33ING2 is a DNA damage-inducible gene that negatively regulates cell proliferation through activation of p53 by enhancing its acetylation. PMID- 11481425 TI - Interactions of Exo1p with components of MutLalpha in Saccharomyces cerevisiae. AB - Previously, we reported evidence suggesting that Saccharomyces cerevisiae MutLalpha, composed of Mlh1p and Pms1p, was a functional member of the gyrase b/Hsp90/MutL (GHL) dimeric ATPase superfamily characterized by highly conserved ATPase domains. Similar to other GHL ATPases, these putative ATPase domains of MutLalpha may be important for the recruitment and/or activation of downstream effectors. One downstream effector candidate is Exo1p, a 5'-3' double stranded DNA exonuclease that has previously been implicated in DNA mismatch repair (MMR). Here we report yeast two-hybrid results suggesting that Exo1p can interact physically with MutLalpha through the Mlh1p subunit. We also report epistasis analysis involving MutLalpha ATPase mutations combined with exo1Delta. One interpretation of our genetic results is that MutLalpha ATPase domains function to direct Exo1p and other functionally redundant exonucleases during MMR. Finally, our results show that much of the increase in spontaneous mutation observed in an exo1Delta strain is REV3-dependent, in turn suggesting that Exo1p is also involved in one or more MMR-independent mutation avoidance pathways. PMID- 11481426 TI - The evolution of methicillin resistance in Staphylococcus aureus: similarity of genetic backgrounds in historically early methicillin-susceptible and -resistant isolates and contemporary epidemic clones. AB - The key genetic component of methicillin resistance, the mecA determinant, is not native to Staphylococcus aureus. Thus, the evolution of methicillin-resistant S. aureus (MRSA) must have begun with the acquisition of the mecA determinant from an unknown heterologous source some time before the first reported appearance of MRSA isolates in clinical specimens in the U.K. and Denmark (in the early 1960s). We compared the genetic backgrounds and phenotypes of a group of methicillin susceptible S. aureus (MSSA) isolates to the properties of MRSA strains isolated in Denmark and the U.K. during the same time period, and also to the genetic profiles of contemporary epidemic clones of MRSA. All early MRSA isolates resembled a large group of the early MSSA blood isolates in phenotypic and genetic properties, including phage group, antibiotype (resistance to penicillin, streptomycin, and tetracycline), pulsed-field gel electrophoresis pattern, and spaA type and multilocus sequence type, strongly suggesting that the early MSSA examined here represented the progeny of a strain that served as one of the first S. aureus recipients of the methicillin-resistance determinant in Europe. The genetic background of this group of early MSSA isolates was also very similar to that of the widely disseminated contemporary "Iberian clone" of MRSA, suggesting that genetic determinants present in early MSSA and essential for some aspects of the epidemicity and/or virulence of these strains may have been retained by this highly successful contemporary MRSA lineage. PMID- 11481427 TI - Spatial-temporal imaging of bacterial infection and antibiotic response in intact animals. AB - We describe imaging the luminance of green fluorescent protein (GFP)-expressing bacteria from outside intact infected animals. This simple, nonintrusive technique can show in great detail the spatial-temporal behavior of the infectious process. The bacteria, expressing the GFP, are sufficiently bright as to be clearly visible from outside the infected animal and recorded with simple equipment. Introduced bacteria were observed in several mouse organs including the peritoneal cavity, stomach, small intestine, and colon. Instantaneous real time images of the infectious process were acquired by using a color charge coupled device video camera by simply illuminating mice at 490 nm. Most techniques for imaging the interior of intact animals may require the administration of exogenous substrates, anesthesia, or contrasting substances and require very long data collection times. In contrast, the whole-body fluorescence imaging described here is fast and requires no extraneous agents. The progress of Escherichia coli-GFP through the mouse gastrointestinal tract after gavage was followed in real-time by whole-body imaging. Bacteria, seen first in the stomach, migrated into the small intestine and subsequently into the colon, an observation confirmed by intravital direct imaging. An i.p. infection was established by i.p. injection of E. coli-GFP. The development of infection over 6 h and its regression after kanamycin treatment were visualized by whole-body imaging. This imaging technology affords a powerful approach to visualizing the infection process, determining the tissue specificity of infection, and the spatial migration of the infectious agents. PMID- 11481428 TI - Kinetic trapping of DNA by transcription factor IIIB. AB - High levels of RNA polymerase III gene transcription are achieved by facilitated recycling of the polymerase on transcription factor IIIB (TFIIIB)-DNA complexes that are stable through multiple rounds of initiation. TFIIIB-DNA complexes in yeast comprise the TATA-binding protein (TBP), the TFIIB-related factor TFIIIB70, and TFIIIB90. The high stability of the TFIIIB-DNA complex is conferred by TFIIIB90 binding to TFIIIB70-TBP-DNA complexes. This stability is thought to result from compound bends introduced in the DNA by TBP and TFIIIB90 and by protein-protein interactions that obstruct DNA dissociation. Here we present biochemical evidence that the high stability of TFIIIB-DNA complexes results from kinetic trapping of the DNA. Thermodynamic analysis shows that the free energies of formation of TFIIIB70-TBP-DNA (DeltaG degrees = -12.10 +/- 0.12 kcal/mol) and TFIIIB-DNA (DeltaG degrees = -11.90 +/- 0.14 kcal/mol) complexes are equivalent whereas a kinetic analysis shows that the half-lives of these complexes (46 +/- 3 min and 95 +/- 6 min, respectively) differ significantly. The differential stability of these isoenergetic complexes demonstrates that TFIIIB90 binding energy is used to drive conformational changes and increase the barrier to complex dissociation. PMID- 11481429 TI - Receptor subtype-specific pronociceptive and analgesic actions of galanin in the spinal cord: selective actions via GalR1 and GalR2 receptors. AB - Galanin is a 29-aa neuropeptide with a complex role in pain processing. Several galanin receptor subtypes are present in dorsal root ganglia and spinal cord with a differential distribution. Here, we describe a generation of a specific galanin R2 (GalR2) agonist, AR-M1896, and its application in studies of a rat neuropathic pain model (Bennett). The results show that in normal rats mechanical and cold allodynia of the hindpaw are induced after intrathecal infusion of low-dose galanin (25 ng per 0.5 microl/h). The same effect is seen with equimolar doses of AR-M1896 or AR-M961, an agonist both at GalR1 and GalR2 receptors. In allodynic Bennett model rats, the mechanical threshold increased dose-dependently after intrathecal injection of a high dose of AR-M961, whereas no effect was observed in the control or AR-M1896 group. No effect of either of the two compounds was observed in nonallodynic Bennett model rats. These data indicate that a low dose of galanin has a nociceptive role at the spinal cord level mediated by GalR2 receptors, whereas the antiallodynic effect of high-dose galanin on neuropathic pain is mediated by the GalR1 receptors. Thus, a selective GalR1 agonist may be used to treat neuropathic pain. PMID- 11481430 TI - Analysis of the chromosome sequence of the legume symbiont Sinorhizobium meliloti strain 1021. AB - Sinorhizobium meliloti is an alpha-proteobacterium that forms agronomically important N(2)-fixing root nodules in legumes. We report here the complete sequence of the largest constituent of its genome, a 62.7% GC-rich 3,654,135-bp circular chromosome. Annotation allowed assignment of a function to 59% of the 3,341 predicted protein-coding ORFs, the rest exhibiting partial, weak, or no similarity with any known sequence. Unexpectedly, the level of reiteration within this replicon is low, with only two genes duplicated with more than 90% nucleotide sequence identity, transposon elements accounting for 2.2% of the sequence, and a few hundred short repeated palindromic motifs (RIME1, RIME2, and C) widespread over the chromosome. Three regions with a significantly lower GC content are most likely of external origin. Detailed annotation revealed that this replicon contains all housekeeping genes except two essential genes that are located on pSymB. Amino acid/peptide transport and degradation and sugar metabolism appear as two major features of the S. meliloti chromosome. The presence in this replicon of a large number of nucleotide cyclases with a peculiar structure, as well as of genes homologous to virulence determinants of animal and plant pathogens, opens perspectives in the study of this bacterium both as a free-living soil microorganism and as a plant symbiont. PMID- 11481431 TI - The complete sequence of the 1,683-kb pSymB megaplasmid from the N2-fixing endosymbiont Sinorhizobium meliloti. AB - Analysis of the 1,683,333-nt sequence of the pSymB megaplasmid from the symbiotic N(2)-fixing bacterium Sinorhizobium meliloti revealed that the replicon has a high gene density with a total of 1,570 protein-coding regions, with few insertion elements and regions duplicated elsewhere in the genome. The only copies of an essential arg-tRNA gene and the minCDE genes are located on pSymB. Almost 20% of the pSymB sequence carries genes encoding solute uptake systems, most of which were of the ATP-binding cassette family. Many previously unsuspected genes involved in polysaccharide biosynthesis were identified and these, together with the two known distinct exopolysaccharide synthesis gene clusters, show that 14% of the pSymB sequence is dedicated to polysaccharide synthesis. Other recognizable gene clusters include many involved in catabolic activities such as protocatechuate utilization and phosphonate degradation. The functions of these genes are consistent with the notion that pSymB plays a major role in the saprophytic competence of the bacteria in the soil environment. PMID- 11481432 TI - Nucleotide sequence and predicted functions of the entire Sinorhizobium meliloti pSymA megaplasmid. AB - The symbiotic nitrogen-fixing soil bacterium Sinorhizobium meliloti contains three replicons: pSymA, pSymB, and the chromosome. We report here the complete 1,354,226-nt sequence of pSymA. In addition to a large fraction of the genes known to be specifically involved in symbiosis, pSymA contains genes likely to be involved in nitrogen and carbon metabolism, transport, stress, and resistance responses, and other functions that give S. meliloti an advantage in its specialized niche. PMID- 11481433 TI - Peptide methionine sulfoxide reductase from Escherichia coli and Mycobacterium tuberculosis protects bacteria against oxidative damage from reactive nitrogen intermediates. AB - Inducible nitric oxide synthase (iNOS) plays an important role in host defense. Macrophages expressing iNOS release the reactive nitrogen intermediates (RNI) nitrite and S-nitrosoglutathione (GSNO), which are bactericidal in vitro at a pH characteristic of the phagosome of activated macrophages. We sought to characterize the active intrabacterial forms of these RNI and their molecular targets. Peptide methionine sulfoxide reductase (MsrA; EC ) catalyzes the reduction of methionine sulfoxide (Met-O) in proteins to methionine (Met). E. coli lacking MsrA were hypersensitive to killing not only by hydrogen peroxide, but also by nitrite and GSNO. The wild-type phenotype was restored by transformation with plasmids encoding msrA from E. coli or M. tuberculosis, but not by an enzymatically inactive mutant msrA, indicating that Met oxidation was involved in the death of these cells. It seemed paradoxical that nitrite and GSNO kill bacteria by oxidizing Met residues when these RNI cannot themselves oxidize Met. However, under anaerobic conditions, neither nitrite nor GSNO was bactericidal. Nitrite and GSNO can both give rise to NO, which may react with superoxide produced by bacteria during aerobic metabolism, forming peroxynitrite, a known oxidant of Met to Met-O. Thus, the findings are consistent with the hypotheses that nitrite and GSNO kill E. coli by intracellular conversion to peroxynitrite, that intracellular Met residues in proteins constitute a critical target for peroxynitrite, and that MsrA can be essential for the repair of peroxynitrite-mediated intracellular damage. PMID- 11481434 TI - Trypanosoma cruzi trans-sialidase: a potent and specific survival factor for human Schwann cells by means of phosphatidylinositol 3-kinase/Akt signaling. AB - Patients infected with Trypanosoma cruzi may remain asymptomatic for decades and show signs of neuroregeneration in the peripheral nervous system (PNS). In the absence of such neuroregeneration, patients may die in part by extensive neuronal destruction in the gastrointestinal tract. Thus, T. cruzi may, despite their invasion of the PNS, directly prevent cell death to keep nerve destruction in check. Indeed, T. cruzi invasion of Schwann cells, their prime target in PNS, suppressed host-cell apoptosis caused by growth-factor deprivation. The trans sialidase (TS) of T. cruzi and the Cys-rich domain of TS reproduced the antiapoptotic activity of the parasites at doses (> or =3.0 nM) comparable or lower than those of bona fide mammalian growth factors. This effect was blocked by LY294002, an inhibitor of phosphatidylinositol 3-kinase (PI3K). TS also activated Akt, a downstream effector of PI3K. Ectopic expression of TS in an unrelated parasite, Leishmania major, turned those parasites into activators of Akt in Schwann cells. In contrast, the Cys-rich domain of TS did not block apoptosis in Schwann cells overexpressing dominant-negative Akt or constitutively active PTEN, a negative regulator of PI3K/Akt signaling. The results demonstrate that T. cruzi, through its TS, triggers the survival of host Schwann cells via the PI3K/Akt pathway, suggesting a role for PI3K/Akt in the pathogenesis of Chagas' disease. PMID- 11481435 TI - Identification of peptides from brain and pituitary of Cpe(fat)/Cpe(fat) mice. AB - Cpe(fat)/Cpe(fat) mice have a naturally occurring point mutation within the carboxypeptidase E gene that inactivates this enzyme, leading to an accumulation of many neuroendocrine peptides containing C-terminal basic residues. These processing intermediates can be readily purified on an anhydrotrypsin affinity resin. Using MS to obtain molecular mass and partial sequence information, more than 100 peptides have been identified. These peptides represent fragments of 16 known secretory pathway proteins, including proenkephalin, proopiomelanocortin, protachykinins A and B, chromogranin A and B, and secretogranin II. Many of the identified peptides represent previously uncharacterized fragments of the precursors. For example, 12 of the 13 chromogranin B-derived peptides found in the present study have not been previously reported. Of these 13 chromogranin B derived peptides, only five contain consensus cleavage sites for prohormone convertases at both the C and N termini. Two distinct chromogranin B-derived peptides result from cleavage at Trp-Trp bonds, a site not typically associated with neuropeptide processing. An RIA was used to confirm that one of these peptides, designated WE-15, exists in wild-type mouse brain, thus validating the approach to identify peptides in Cpe(fat)/Cpe(fat) mice. These "orphan" peptides are candidate ligands for orphan G protein-coupled receptors. In addition, the general technique of using affinity chromatography to isolate endogenous substrates from a mutant organism lacking an enzyme should be applicable to a wide range of enzyme-substrate systems. PMID- 11481436 TI - Functional activity and role of cation-efflux family members in Ni hyperaccumulation in Thlaspi goesingense. AB - The ability of Thlaspi goesingense to hyperaccumulate Ni seems to be governed in part by enhanced accumulation of Ni within leaf vacuoles. We have characterized genes from T. goesingense encoding putative vacuolar metal ion transport proteins, termed metal tolerance proteins (TgMTPs). These proteins contain all of the features of cation-efflux family members, and evidence indicates they are derived from a single genomic sequence (TgMTP1) that gives rise to an unspliced (TgMTP1t1) and a spliced (TgMTP1t2) transcript. Heterologous expression of these transcripts in yeast lacking the TgMTP1 orthologues COT1 and ZRC1 complements the metal sensitivity of these yeast strains, suggesting that TgMTP1s are able to transport metal ions into the yeast vacuole in a manner similar to COT1 and ZRC1. The unspliced and spliced TgMTP1 variants differ within a histidine-rich putative metal-binding domain, and these sequence differences are reflected as alterations in the metal specificities of these metal ion transporters. When expressed in yeast, TgMTP1t1 confers the highest level of tolerance to Cd, Co, and Zn, whereas TgMTP1t2 confers the highest tolerance to Ni. TgMTP1 transcripts are highly expressed in T. goesingense compared with orthologues in the nonaccumulators Arabidopsis thaliana, Thlaspi arvense, and Brassica juncea. We propose that the high-level expression of TgMTP1 in T. goesingense accounts for the enhanced ability of this hyperaccumulator to accumulate metal ions within shoot vacuoles. PMID- 11481437 TI - UV irradiation augments lymphoid malignancies in mice with one functional copy of wild-type p53. AB - Epidemiological studies have suggested an association between exposure to solar UV radiation and the incidence of lymphoid malignancies, which has increased substantially worldwide during the last two decades. Findings from animal studies have raised the question of whether UV radiation might influence the development of lymphoid malignancies by means of its immunosuppressive effect. In this study, we examined the effect of UV irradiation on the development of lymphoid malignancies in mice with no or only one functional copy of p53. Mice that lack both copies of p53 spontaneously develop high frequency of lymphoid malignancies in the thymus and spleen. p53 heterozygous mice with only one copy of the wild type allele also develop lymphoid malignancies, but with a much lower frequency and a long latent period. In our study using mice of the C57BL/6 background, only one of the unirradiated mice lacking one copy of p53 (p53(+/-)) spontaneously developed a lymphoid tumor (6%), whereas 88% of UV-irradiated p53(+/-) mice developed lymphoid tumors in the spleen or liver. None of the control or UV irradiated p53 wild-type mice developed lymphoid tumors during the 60-week observation period. Both UV-irradiated and unirradiated mice lacking both copies of p53 (p53(-/-)) rapidly developed thymic lymphomas and/or lymphoid tumors in spleen or liver. All of the lymphoid tumors tested were of T cell type. The immune responses of the mice to contact sensitization were identical and were suppressed to the same extent by UV irradiation regardless of the genotype. These results indicate that differences in immune reactivity do not account for the different effects of UV radiation on lymphoid malignancies and, in addition, that p53 is not required for generation of T cell-mediated immunity. Interestingly, whereas p53 mutations or loss of heterozygosity did not account for the accelerated development of lymphoid tumors in UV-irradiated p53(+/-) mice, deletions in the p16(INK4a) gene were quite common. These data provide the experimental evidence that UV irradiation induces lymphoid neoplasms in genetically susceptible mice and support the hypothesis that extensive sunlight exposure contributes to the induction of lymphoma in humans. PMID- 11481438 TI - HIN-1, a putative cytokine highly expressed in normal but not cancerous mammary epithelial cells. AB - To identify molecular alterations implicated in the initiating steps of breast tumorogenesis, we compared the gene expression profiles of normal and ductal carcinoma in situ (DCIS) mammary epithelial cells by using serial analysis of gene expression (SAGE). Through the pair-wise comparison of normal and DCIS SAGE libraries, we identified several differentially expressed genes. Here, we report the characterization of one of these genes, HIN-1 (high in normal-1). HIN-1 expression is significantly down regulated in 94% of human breast carcinomas and in 95% of preinvasive lesions, such as ductal and lobular carcinoma in situ. This decrease in HIN-1 expression is accompanied by hypermethylation of its promoter in the majority of breast cancer cell lines (>90%) and primary tumors (74%). HIN 1 is a putative cytokine with no significant homology to known proteins. Reintroduction of HIN-1 into breast cancer cells inhibits cell growth. These results indicate that HIN-1 is a candidate tumor suppressor gene that is inactivated at high frequency in the earliest stages of breast tumorogenesis. PMID- 11481439 TI - Three-dimensional structure of a mammalian thioredoxin reductase: implications for mechanism and evolution of a selenocysteine-dependent enzyme. AB - Thioredoxin reductases (TrxRs) from mammalian cells contain an essential selenocysteine residue in the conserved C-terminal sequence Gly-Cys-SeCys-Gly forming a selenenylsulfide in the oxidized enzyme. Reduction by NADPH generates a selenolthiol, which is the active site in reduction of Trx. The three-dimensional structure of the SeCys498Cys mutant of rat TrxR in complex with NADP(+) has been determined to 3.0-A resolution by x-ray crystallography. The overall structure is similar to that of glutathione reductase (GR), including conserved amino acid residues binding the cofactors FAD and NADPH. Surprisingly, all residues directly interacting with the substrate glutathione disulfide in GR are conserved despite the failure of glutathione disulfide to act as a substrate for TrxR. The 16 residue C-terminal tail, which is unique to mammalian TrxR, folds in such a way that it can approach the active site disulfide of the other subunit in the dimer. A model of the complex of TrxR with Trx suggests that electron transfer from NADPH to the disulfide of the substrate is possible without large conformational changes. The C-terminal extension typical of mammalian TrxRs has two functions: (i) it extends the electron transport chain from the catalytic disulfide to the enzyme surface, where it can react with Trx, and (ii) it prevents the enzyme from acting as a GR by blocking the redox-active disulfide. Our results suggest that mammalian TrxR evolved from the GR scaffold rather than from its prokaryotic counterpart. This evolutionary switch renders cell growth dependent on selenium. PMID- 11481440 TI - Regulation of the transcriptional coactivator PGC-1 via MAPK-sensitive interaction with a repressor. AB - Mechanisms and signals that regulate transcriptional coactivators are still largely unknown. Here we provide genetic evidence for a repressor that interacts with and regulates the nuclear receptor coactivator PGC-1. Association with the repressor requires a PGC-1 protein interface that is similar to the one used by nuclear receptors. Removal of the repressor enhances PGC-1 coactivation of steroid hormone responses. We also provide evidence that interaction of the repressor with PGC-1 is regulated by mitogen-activated protein kinase (MAPK) signaling. Activation of the MAPK p38 enhances the activity of wild-type PGC-1 but not of a PGC-1 variant that no longer interacts with the repressor. Finally, p38 activation enhances steroid hormone response in a PGC-1-dependent manner. Our data suggest a model where the repressor and nuclear receptors compete for recruiting PGC-1 to an inactive and active state, respectively. Extracellular signals such as nuclear receptor ligands or activators of the MAPK p38 can shift the equilibrium between the two states. PMID- 11481441 TI - Activation and autoregulation of DNA-PK from structured single-stranded DNA and coding end hairpins. AB - DNA-dependent protein kinase (DNA-PK) acts through an essential relationship with DNA to participate in the regulation of multiple cellular processes. Yet the role of DNA as a cofactor in kinase activity remains to be completely elucidated. For example, although DNA-PK activity appears to be required for the resolution of hairpin coding ends in variable diversity joining recombination, kinase activity remains to be demonstrated from hairpin ends or other DNA structures. In the present study we report that DNA-PK is strongly activated from hairpin ends and structured single-stranded DNA, but that the phosphorylation of many heterologous substrates is blocked efficiently by inactivation of the kinase through autophosphorylation. However, substrates that bound efficiently to single stranded DNA such as p53 and replication protein A were efficiently phosphorylated by DNA-PK from structured DNA. DNA-PK also was found to be active toward heterologous substrates from hairpin ends on double-stranded DNA under conditions where autophosphorylation was minimized. These results suggest that the role of DNA-PK in resolving coding end hairpins is likely to be enzymatic rather than structural, expand understanding of how DNA-PK binding to structured DNA relates to enzyme activity, and suggest a mechanism for autoregulatory control of its kinase activity in the cell. PMID- 11481442 TI - Differential effects of p19(Arf) and p16(Ink4a) loss on senescence of murine bone marrow-derived preB cells and macrophages. AB - Establishment of cell lines from primary mouse embryo fibroblasts depends on loss of either the Arf tumor suppressor or its downstream target, the p53 transcription factor. Mouse p19(Arf) is encoded by the Ink4a-Arf locus, which also specifies a second tumor suppressor protein, the cyclin D-dependent kinase inhibitor p16(Ink4a). We surveyed bone marrow-derived cells from wild-type, Ink4a Arf-null, or Arf-null mice for their ability to bypass senescence during continuous passage in culture. Unlike preB cells from wild-type mice, those from mice lacking Arf alone could be propagated indefinitely when placed onto stromal feeder layers engineered to produce IL-7. The preB cell lines remained diploid and IL-7-dependent and continued to express elevated levels of p16(Ink4a). By contrast, Arf-null bone marrow-derived macrophages that depend on colony stimulating factor-1 for proliferation and survival in culture initially grew at a slow rate but gave rise to rapidly and continuously growing, but still growth factor-dependent, variants that ceased to express p16(Ink4a). Wild-type bone marrow-derived macrophages initially expressed both p16(Ink4a) and p19(Arf) but exhibited an extended life span when p16(Ink4a) expression was extinguished. In all cases, gene silencing was accompanied by methylation of the Ink4a promoter. Therefore, whereas Arf loss alone appears to be the major determinant of establishment of murine fibroblast and preB cell lines in culture, p16(Ink4a) provides an effective barrier to immortalization of bone marrow-derived macrophages. PMID- 11481443 TI - Enhanced synthesis of choline and glycine betaine in transgenic tobacco plants that overexpress phosphoethanolamine N-methyltransferase. AB - Choline (Cho) is the precursor of the osmoprotectant glycine betaine and is itself an essential nutrient for humans. Metabolic engineering of Cho biosynthesis in plants could therefore enhance both their resistance to osmotic stresses (drought and salinity) and their nutritional value. The key enzyme of the plant Cho-synthesis pathway is phosphoethanolamine N-methyltransferase, which catalyzes all three of the methylations required to convert phosphoethanolamine to phosphocholine. We show here that overexpressing this enzyme in transgenic tobacco increased the levels of phosphocholine by 5-fold and free Cho by 50-fold without affecting phosphatidylcholine content or growth. Moreover, the expanded Cho pool led to a 30-fold increase in synthesis of glycine betaine via an engineered glycine betaine pathway. Supplying the transgenics with the Cho precursor ethanolamine (EA) further enhanced Cho levels even though the supplied EA was extensively catabolized. These latter results establish that there is further scope for improving Cho synthesis by engineering an increased endogenous supply of EA and suggest that this could be achieved by enhancing EA synthesis and/or by suppressing its degradation. PMID- 11481444 TI - A functional role for Tsix transcription in blocking Xist RNA accumulation but not in X-chromosome choice. AB - In female mammals, up-regulation of Xist triggers X-chromosome inactivation in cis. Up-regulation is inhibited by sequences 3' to Xist contained within the antisense locus, Tsix. Inhibition could depend on transcription of Tsix and/or on DNA elements therein. Here we test the role of Tsix transcription by augmenting the duration and strength of Tsix expression. We find that Tsix hypertranscription is sufficient to block Xist RNA accumulation in a cis-limited manner. We propose that Tsix transcription is necessary to restrict Xist activity on the future active X and, conversely, that Tsix repression is required for Xist RNA accumulation on the future inactive X. We also find that Tsix hypertranscription does not affect X-chromosome choice. Thus, choice is mediated by elements within Tsix that are independent of promoter activity. PMID- 11481445 TI - Dimerization of a selectin and its ligand stabilizes cell rolling and enhances tether strength in shear flow. AB - Selectins mediate rolling of leukocytes by rapid formation and dissociation of selectin-ligand bonds, which are assumed to require high mechanical strength to prevent premature dissociation by the forces applied in shear flow. This assumption is based largely on the observation that increasing wall shear stress increases only modestly the dissociation of transient leukocyte tethers on very low selectin densities. P-selectin binds to the N-terminal region of P-selectin glycoprotein ligand-1 (PSGL-1), a mucin on leukocytes. Both PSGL-1 and P-selectin are extended homodimers. We perfused transfected cells expressing wild-type dimeric PSGL-1 or a chimeric monomeric form of PSGL-1 on immobilized dimeric or monomeric forms of P-selectin. Cells expressing dimeric or monomeric PSGL-1 tethered to P-selectin at equivalent rates. However, cells expressing dimeric PSGL-1 established more stable rolling adhesions, which were more shear resistant and exhibited less fluctuation in rolling velocities. On low densities of dimeric P-selectin, increasing wall shear stress more rapidly increased transient tether dissociation of cells expressing monomeric PSGL-1 than dimeric PSGL-1. Tether dissociation on low densities of monomeric P-selectin was even more shear sensitive. We conclude that dimerization of both PSGL-1 and P-selectin stabilizes tethering and rolling, probably by increasing rebinding within a bond cluster. Because transient tethers may have more than one bond, the mechanical strength of selectin-ligand bonds is likely to be lower than initially estimated. Tether strength may rely more on bond clusters to distribute applied force. PMID- 11481446 TI - Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. AB - Short interfering RNAs (siRNAs) are double-stranded RNAs of approximately 21-25 nucleotides that have been shown to function as key intermediaries in triggering sequence-specific RNA degradation during posttranscriptional gene silencing in plants and RNA interference in invertebrates. siRNAs have a characteristic structure, with 5'-phosphate/3'-hydroxyl ends and a 2-base 3' overhang on each strand of the duplex. In this study, we present data that synthetic siRNAs can induce gene-specific inhibition of expression in Caenorhabditis elegans and in cell lines from humans and mice. In each case, the interference by siRNAs was superior to the inhibition of gene expression mediated by single-stranded antisense oligonucleotides. The siRNAs seem to avoid the well documented nonspecific effects triggered by longer double-stranded RNAs in mammalian cells. These observations may open a path toward the use of siRNAs as a reverse genetic and therapeutic tool in mammalian cells. PMID- 11481447 TI - A soluble class II cytokine receptor, IL-22RA2, is a naturally occurring IL-22 antagonist. AB - IL-22 is an IL-10 homologue that binds to and signals through the class II cytokine receptor heterodimer IL-22RA1/CRF2-4. IL-22 is produced by T cells and induces the production of acute-phase reactants in vitro and in vivo, suggesting its involvement in inflammation. Here we report the identification of a class II cytokine receptor designated IL-22RA2 (IL-22 receptor-alpha 2) that appears to be a naturally expressed soluble receptor. IL-22RA2 shares amino acid sequence homology with IL-22RA1 (also known as IL-22R, zcytor11, and CRF2-9) and is physically adjacent to IL-20Ralpha and IFN-gammaR1 on chromosome 6q23.3-24.2. We demonstrate that IL-22RA2 binds specifically to IL-22 and neutralizes IL-22 induced proliferation of BaF3 cells expressing IL-22 receptor subunits. IL-22RA2 mRNA is highly expressed in placenta and spleen by Northern blotting. PCR analysis using RNA from various tissues and cell lines showed that IL-22RA2 was expressed in a range of tissues, including those in the digestive, female reproductive, and immune systems. In situ hybridization revealed the dominant cell types expressing IL-22RA2 were mononuclear cells and epithelium. Because IL 22 induces the expression of acute phase reactants, IL-22RA2 may play an important role as an IL-22 antagonist in the regulation of inflammatory responses. PMID- 11481448 TI - Endothelial cell-specific knockout of connexin 43 causes hypotension and bradycardia in mice. AB - Connexin 43 (Cx43) is a protein expressed in a variety of mammalian tissues. However, the lack of specific blockers and the absence of known genetic mutants have hampered the investigation of the function of this protein. Cx43-null mice die shortly after birth, thus preventing functional studies in vivo. Here, we report the generation and characterization of a vascular endothelial cell specific deletion of the Cx43 gene (VEC Cx43 KO) in mice by using the loxP/Cre system. Using homologous recombination, a mouse line was created carrying loxP sites flanking exon 2 of the Cx43 gene ("floxed" mice). To produce cell specific deletion of the Cx43 gene, these mice were crossed with animals from a line carrying the Tie 2-Cre transgene. The homozygous VEC Cx43 KO mice survived to maturity. However, they were hypotensive and bradycardic when compared with heterozygous VEC Cx43 KO mice, or to the floxed Cx43 gene mice. The hypotension was associated with marked elevation of plasma nitric oxide (NO) levels as well as elevated plasma angiotensin (Ang) I and II. We hypothesize that endothelial cell Cx43 plays a key role in the formation and/or action of NO, and that the elevation of Ang II is a secondary event. The specific cellular basis for the hypotension remains to be established, but our findings support the idea that endothelial Cx43 gap junctions are involved in maintaining normal vascular function; moreover, these animals provide the opportunity to determine more clearly the role of endothelial Cx43 in vascular development and homeostasis. PMID- 11481449 TI - Sexual dimorphism among bovine embryos in their ability to make the transition to expanded blastocyst and in the expression of the signaling molecule IFN-tau. AB - IFN-tau is a secretory product of trophectoderm of cattle, sheep, and their relatives and is expressed for a few days in early pregnancy after the blastocyst first forms. It serves to alert the mother that she is pregnant. A delayed or less than robust IFN-tau signal is a likely cause of embryonic loss. Here we have determined whether blastocyst production of IFN-tau, which is encoded by a cluster of genes on chromosome 9, differs between the sexes in cattle, as assessed by culture of in vitro-derived embryos on two different media, one complex (tissue culture medium 199 supplemented with serum) with coculture support, the other relatively simple (synthetic oviductal fluid plus albumin). With both media, female blastocysts produced approximately double the amount of IFN-tau as males, regardless of such variables as oocyte batch, blastocyst quality, hatching, and length of time in culture. However, in either tissue culture medium 199, which contains 5.5 mM d-glucose, or in synthetic oviductal fluid, in the presence but not in the absence of added glucose, significantly fewer female than male embryos were able to progress from the morula/early blastocyst stage to more advanced stages of development. It is possible that the differences between male and female embryos both in their production of IFN-tau and in their ability to progress in development in glucose-rich media are manifestations of phenomena that occur in vivo and provide plasticity in embryo selection during early pregnancy. PMID- 11481450 TI - Old World sources of the first New World human inhabitants: a comparative craniofacial view. AB - Human craniofacial data were used to assess the similarities and differences between recent and prehistoric Old World samples, and between these samples and a similar representation of samples from the New World. The data were analyzed by the neighbor-joining clustering procedure, assisted by bootstrapping and by canonical discriminant analysis score plots. The first entrants to the Western Hemisphere of maybe 15,000 years ago gave rise to the continuing native inhabitants south of the U.S.-Canadian border. These show no close association with any known mainland Asian population. Instead they show ties to the Ainu of Hokkaido and their Jomon predecessors in prehistoric Japan and to the Polynesians of remote Oceania. All of these also have ties to the Pleistocene and recent inhabitants of Europe and may represent an extension from a Late Pleistocene continuum of people across the northern fringe of the Old World. With roots in both the northwest and the northeast, these people can be described as Eurasian. The route of entry to the New World was at the northwestern edge. In contrast, the Inuit (Eskimo), the Aleut, and the Na-Dene speakers who had penetrated as far as the American Southwest within the last 1,000 years show more similarities to the mainland populations of East Asia. Although both the earlier and later arrivals in the New World show a mixture of traits characteristic of the northern edge of Old World occupation and the Chinese core of mainland Asia, the proportion of the latter is greater for the more recent entrants. PMID- 11481451 TI - Polar targeting of Shigella virulence factor IcsA in Enterobacteriacae and Vibrio. AB - Asymmetric localization is key to the proper function of certain prokaryotic proteins important to virulence, chemotaxis, cell division, development, motility, and adhesion. Shigella IcsA is localized to the old pole of the bacterium, where it mediates assembly of an actin tail inside infected mammalian cells. IcsA (VirG) is essential to Shigella intracellular motility and virulence. We used translational fusions between portions of IcsA and the green fluorescent protein (GFP) to determine the regions of IcsA that are necessary and sufficient for its targeting to the bacterial old pole. An IcsA-GFP fusion that lacks a signal peptide localized to the old pole, indicating that signal peptide-mediated secretion is not required for polar localization. Two regions within IcsA were required for localization of an IcsA-GFP fusion to the old pole. Further characterization of these regions indicated that amino acids 1-104 and 507-620 were each independently sufficient for polar localization. Finally, when expressed in Escherichia coli, Salmonella typhimurium, Yersinia pseudotuberculosis, and Vibrio cholerae, each of the two targeting regions localized to the pole, indicating that the mechanism of polar targeting used by IcsA is present generally among Enterobacteriacae and Vibrio. PMID- 11481452 TI - Human hepatitis C virus NS5A protein alters intracellular calcium levels, induces oxidative stress, and activates STAT-3 and NF-kappa B. AB - The nonstructural protein 5A (NS5A) encoded by the human hepatitis C virus RNA genome is shown here to induce the activation of NF-kappaB and STAT-3 transcription factors from its cytoplasmic residence via oxidative stress. NS5A causes the disturbance of intracellular calcium. Ca2+ signaling triggers the elevation of reactive oxygen species in mitochondria, leading to the translocation of NF-kappaB and STAT-3 into the nucleus. Evidence is presented for the constitutive activation of STAT-3 by NS5A. In the presence of antioxidants [pyrrolidine dithiocarbamate (PDTC), N-acetyl l-cysteine (NAC)] or Ca2+ chelators (EGTA-AM, TMB-8), NS5A-induced activation of NF-kappaB and STAT-3 was eliminated. These results provide an insight into the mechanism by which NS5A can alter intracellular events relevant to liver pathogenesis associated with the viral infection. PMID- 11481453 TI - Antisense DNAs as multisite genomic modulators identified by DNA microarray. AB - Antisense oligodeoxynucleotides can selectively block disease-causing genes, and cancer genes have been chosen as potential targets for antisense drugs to treat cancer. However, nonspecific side effects have clouded the true antisense mechanism of action and hampered clinical development of antisense therapeutics. Using DNA microarrays, we have conducted a systematic characterization of gene expression in cells exposed to antisense, either exogenously or endogenously. Here, we show that in a sequence-specific manner, antisense targeted to protein kinase A RIalpha alters expression of the clusters of coordinately expressed genes at a specific stage of cell growth, differentiation, and activation. The genes that define the proliferation-transformation signature are down-regulated, whereas those that define the differentiation-reverse transformation signature are up-regulated in antisense-treated cancer cells and tumors, but not in host livers. In this differentiation signature, the genes showing the highest induction include genes for the G proteins Rap1 and Cdc42. The expression signature induced by the exogenously supplied antisense oligodeoxynucleotide overlaps strikingly with that induced by endogenous antisense gene overexpression. Defining antisense DNAs on the basis of their effects on global gene expression can lead to identification of clinically relevant antisense therapeutics and can identify which molecular and cellular events might be important in complex biological processes, such as cell growth and differentiation. PMID- 11481454 TI - Structure of the gene 2.5 protein, a single-stranded DNA binding protein encoded by bacteriophage T7. AB - The gene 2.5 protein (gp2.5) of bacteriophage T7 is a single-stranded DNA (ssDNA) binding protein that has essential roles in DNA replication and recombination. In addition to binding DNA, gp2.5 physically interacts with T7 DNA polymerase and T7 primase-helicase during replication to coordinate events at the replication fork. We have determined a 1.9-A crystal structure of gp2.5 and show that it has a conserved OB-fold (oligosaccharide/oligonucleotide binding fold) that is well adapted for interactions with ssDNA. Superposition of the OB-folds of gp2.5 and other ssDNA binding proteins reveals a conserved patch of aromatic residues that stack against the bases of ssDNA in the other crystal structures, suggesting that gp2.5 binds to ssDNA in a similar manner. An acidic C-terminal extension of the gp2.5 protein, which is required for dimer formation and for interactions with the T7 DNA polymerase and the primase-helicase, appears to be flexible and may act as a switch that modulates the DNA binding affinity of gp2.5. PMID- 11481455 TI - Brain glucose utilization in mice with a targeted mutation in the thyroid hormone alpha or beta receptor gene. AB - Brain glucose utilization is markedly depressed in adult rats made cretinous after birth. To ascertain which subtype of thyroid hormone (TH) receptors, TRalpha1 or TRbeta, is involved in the regulation of glucose utilization during brain development, we used the 2-[(14)C]deoxyglucose method in mice with a mutation in either their TRalpha or TRbeta gene. A C insertion produced a frameshift mutation in their carboxyl terminus. These mutants lacked TH binding and transactivation activities and exhibited potent dominant negative activity. Glucose utilization in the homozygous TRbetaPV mutant mice and their wild-type siblings was almost identical in 19 brain regions, whereas it was markedly reduced in all brain regions of the heterozygous TRalpha1PV mice. These suggest that the alpha1 receptor mediates the TH effects in brain. Inasmuch as local cerebral glucose utilization is closely related to local synaptic activity, we also examined which thyroid hormone receptor is involved in the expression of synaptotagmin-related gene 1 (Srg1), a TH-positively regulated gene involved in the formation and function of synapses [Thompson, C. C. (1996) J. Neurosci. 16, 7832-7840]. Northern analysis showed that Srg1 expression was markedly reduced in the cerebellum of TRalpha(PV/+) mice but not TRbeta(PV/PV) mice. These results show that the same receptor, TRalpha1, is involved in the regulation by TH of both glucose utilization and Srg1 expression. PMID- 11481456 TI - Ethanol hypersensitivity and olfactory discrimination defect in mice lacking a homolog of Drosophila neuralized. AB - Neurogenic genes in the Notch receptor-mediated signaling pathway play important roles in neuronal cell fate specification as well as neuronal differentiation. The Drosophila neuralized gene is one of the neurogenic genes. We have cloned a mouse homolog of Drosophila neuralized, m-neu1, and found that the m-neu1 transcript is expressed in differentiated neurons. Mice deficient for m-neu1 are viable and morphologically normal, but exhibit specific defects in olfactory discrimination and hypersensitivity to ethanol. These findings reveal an essential role of m-neu1 in ensuring proper processing of certain information in the adult brain. PMID- 11481457 TI - SMAD4 mutations in colorectal cancer probably occur before chromosomal instability, but after divergence of the microsatellite instability pathway. AB - Loss of chromosome 18q21 is well documented in colorectal cancer, and it has been suggested that this loss targets the DCC, DPC4/SMAD4, and SMAD2 genes. Recently, the importance of SMAD4, a downstream regulator in the TGF-beta signaling pathway, in colorectal cancer has been highlighted, although the frequency of SMAD4 mutations appears much lower than that of 18q21 loss. We set out to investigate allele loss, mutations, protein expression, and cytogenetics of chromosome 18 copy number in a collection of 44 colorectal cancer cell lines of known status with respect to microsatellite instability (MSI). Fourteen of thirty two MSI(-) lines showed loss of SMAD4 protein expression; usually, one allele was lost and the other was mutated in one of a number of ways, including deletions of various sizes, splice site changes, and missense and nonsense point mutations (although no frameshifts). Of the 18 MSI(-) cancers with retained SMAD4 expression, four harbored missense mutations in the 3' part of the gene and showed allele loss. The remaining 14 MSI(-) lines had no detectable SMAD4 mutation, but all showed allele loss at SMAD4 and/or DCC. SMAD4 mutations can therefore account for about 50-60% of the 18q21 allele loss in colorectal cancer. No MSI(+) cancer showed loss of SMAD4 protein or SMAD4 mutation, and very few had allelic loss at SMAD4 or DCC, although many of these MSI(+) lines did carry TGFBIIR changes. Although SMAD4 mutations have been associated with late-stage or metastatic disease, our combined molecular and cytogenetic data best fit a model in which SMAD4 mutations occur before colorectal cancers become aneuploid/polyploid, but after the MSI(+) and MSI(-) pathways diverge. Thus, MSI(+) cancers may diverge first, followed by CIN(+) (chromosomal instability) cancers, leaving other cancers to follow a CIN(-)MSI(-) pathway. PMID- 11481458 TI - Molecular cloning of Porimin, a novel cell surface receptor mediating oncotic cell death. AB - Anti-Porimin (Pro-oncosis receptor inducing membrane injury) mAb mediates oncosis like cell death in Jurkat cells. Porimin cDNA was isolated from a Jurkat cell cDNA library by COS cell-expression cloning. The 3,337-bp cDNA has an ORF of 567 bp, encoding a type I transmembrane protein of 189 amino acids. The extracellular domain of Porimin contains many O-linked and seven N-linked glycosylation sites that define it as a new member of the mucin family. COS7 and 293 cells transiently transfected with Porimin cDNA were specifically recognized by anti Porimin Ab in cell staining and immunoblotting experiments. When expressed in Jurkat cells, a His-tagged Porimin cDNA construct resulted in the generation of a specific 110-kDa-size protein that matched the molecular mass of the endogenous Porimin protein. Crosslinking of the Porimin receptor expressed on COS7 transfectants resulted in the loss of cell membrane integrity and cell death as measured by the leakage of intracellular lactate dehydrogenase. Both COS7 and 293 cells expressing transfected Porimin at a relatively high level lost their ability to adhere to culture dishes, suggesting a role for Porimin in cell adhesion. The Porimin gene was mapped to human chromosome 11q22.1 and is composed of four exons spanning 133 kb of genomic DNA. PMID- 11481459 TI - Treasures in the attic: rolling circle transposons discovered in eukaryotic genomes. PMID- 11481460 TI - Progress in plant metabolic engineering. PMID- 11481461 TI - Mimicry: an interface between psychology and evolution. PMID- 11481462 TI - Abeta immunization: moving Abeta peptide from brain to blood. PMID- 11481463 TI - Progress in cancer vaccines by enhanced self-presentation. PMID- 11481464 TI - Functional expression of a Drosophila odor receptor. PMID- 11481465 TI - Detergent-phospholipid mixed micelles with a crystalline phospholipid core. AB - An unusual micelle was discovered in mixtures of the nonionic detergent octaethyleneglycol-mono-n-dodecylether with disaturated phospholipids such as 1,2 dimyristoyl-sn-glycero-3-phosphocholine or 1,2-dipalmitoyl-sn-glycero-3 phosphocholine in water. These mixtures undergo a structural transition upon cooling through the chain-melting temperatures of the respective phospholipids, resulting in the formation of mixed micelles. Structural features of the micellar particles were studied here by synchrotron x-ray scattering. The translucent micellar solutions showed characteristic wide-angle reflections that were attributed to ordered hydrocarbon chains, whereas the absence of small-angle x ray reflections indicated that there is no long-range order in these mixtures. The presence of ordered phospholipid acyl chains was confirmed by differential scanning calorimetry and isothermal titration calorimetry. The endothermic differential scanning calorimetry signals observed in the up-scan mode were tentatively ascribed to chain melting and mixing of the components. Isothermal titration of the mixed-micellar solutions into an excess of the detergent octaethyleneglycol-mono-n-dodecylether resulted in sudden uptake of the latent heat by the gel-state phospholipids. The heat uptake per mol of phospholipid decreased with increasing detergent/phospholipid molar ratio. A simple geometric model is presented assuming that the dominating particle species in the mixtures is a discoidal phospholipid aggregate with ordered acyl chains, surrounded by a toroidal detergent hoop. The model implies that the fraction of ordered phospholipid chains decreases with increasing detergent/phospholipid molar ratio, in agreement with the calorimetric results and high-resolution NMR spectroscopy. PMID- 11481466 TI - Image metrics in the statistical analysis of DNA microarray data. AB - DNA microarrays represent an important new method for determining the complete expression profile of a cell. In "spotted" microarrays, slides carrying spots of target DNA are hybridized to fluorescently labeled cDNA from experimental and control cells and the arrays are imaged at two or more wavelengths. In this paper, we perform statistical analysis on images of microarrays and show that quantitating the amount of fluorescent DNA bound to microarrays is subject to considerable uncertainty because of large and small-scale intensity fluctuations within spots, nonadditive background, and fabrication artifacts. Pixel-by-pixel analysis of individual spots can be used to estimate these sources of error and establish the precision and accuracy with which gene expression ratios are determined. Simple weighting schemes based on these estimates are effective in improving significantly the quality of microarray data as it accumulates in a multiexperiment database. We propose that error estimates from image-based metrics should be one component in an explicitly probabilistic scheme for the analysis of DNA microarray data. PMID- 11481467 TI - DNA microarray reveals changes in gene expression of shear stressed human umbilical vein endothelial cells. AB - Using DNA microarray screening (GeneFilter 211, Research Genetics, Huntsville, AL) of mRNA from primary human umbilical vein endothelial cells (HUVEC), we identified 52 genes with significantly altered expression under shear stress [25 dynes/cm(2) for 6 or 24 h (1 dyne = 10 microN), compared with matched stationary controls]; including several genes not heretofore recognized to be shear stress responsive. We examined mRNA expression of nine genes by Northern blot analysis, which confirmed the results obtained on DNA microarrays. Thirty-two genes were up regulated (by more than 2-fold), the most enhanced being cytochromes P450 1A1 and 1B1, zinc finger protein EZF/GKLF, glucocorticoid-induced leucine zipper protein, argininosuccinate synthase, and human prostaglandin transporter. Most dramatically decreased (by more than 2-fold) were connective tissue growth factor, endothelin-1, monocyte chemotactic protein-1, and spermidine/spermine N1 acetyltransferase. The changes observed suggest several potential mechanisms for increased NO production under shear stress in endothelial cells. PMID- 11481468 TI - Restructuring of an RNA polymerase holoenzyme elongation complex by lambdoid phage Q proteins. AB - The structure of an intermediate in the initiation to elongation transition of Escherichia coli RNA polymerase has been visualized through region-specific DNA cleavage by the hydroxyl radical reagent FeBABE. FeBABE was tethered to specific sites of the final sigma(70) subunit and incorporated into two specialized paused elongation complexes that obligatorily retain the final sigma(70) initiation subunit and are targets for modification by lambdoid phage late gene antiterminators. The FeBABE cleavage pattern reveals structures similar to open complex, except for notable changes to region 3 of final sigma(70) that might reflect the presence of stably bound transcript. Binding of the antiterminator protein Q displaces the reactivity of FeBABE conjugated to region 4 of final sigma(70), suggesting that final sigma(70) subunit rearrangement is a step in conversion of RNAP to the antiterminating form. PMID- 11481469 TI - Structural insights into the catalytic mechanism of a family 18 exo-chitinase. AB - Chitinase B (ChiB) from Serratia marcescens is a family 18 exo-chitinase whose catalytic domain has a TIM-barrel fold with a tunnel-shaped active site. We have solved structures of three ChiB complexes that reveal details of substrate binding, substrate-assisted catalysis, and product displacement. The structure of an inactive ChiB mutant (E144Q) complexed with a pentameric substrate (binding in subsites -2 to +3) shows closure of the "roof" of the active site tunnel. It also shows that the sugar in the -1 position is distorted to a boat conformation, thus providing structural evidence in support of a previously proposed catalytic mechanism. The structures of the active enzyme complexed to allosamidin (an analogue of a proposed reaction intermediate) and of the active enzyme soaked with pentameric substrate show events after cleavage of the glycosidic bond. The latter structure shows reopening of the roof of the active site tunnel and enzyme assisted product displacement in the +1 and +2 sites, allowing a water molecule to approach the reaction center. Catalysis is accompanied by correlated structural changes in the core of the TIM barrel that involve conserved polar residues whose functions were hitherto unknown. These changes simultaneously contribute to stabilization of the reaction intermediate and alternation of the pKa of the catalytic acid during the catalytic cycle. PMID- 11481470 TI - Substrate conformational transitions in the active site of chorismate mutase: their role in the catalytic mechanism. AB - Chorismate mutase acts at the first branch-point of aromatic amino acid biosynthesis and catalyzes the conversion of chorismate to prephenate. The results of molecular dynamics simulations of the substrate in solution and in the active site of chorismate mutase are reported. Two nonreactive conformers of chorismate are found to be more stable than the reactive pseudodiaxial chair conformer in solution. It is shown by QM/MM molecular dynamics simulations, which take into account the motions of the enzyme, that when these inactive conformers are bound to the active site, they are rapidly converted to the reactive chair conformer. This result suggests that one contribution of the enzyme is to bind the more prevalent nonreactive conformers and transform them into the active form in a step before the chemical reaction. The motion of the reactive chair conformer in the active site calculated by using the QM/MM potential generates transient structures that are closer to the transition state than is the stable CHAIR conformer. PMID- 11481472 TI - The Calpha ---H...O hydrogen bond: a determinant of stability and specificity in transmembrane helix interactions. AB - The Calpha---H...O hydrogen bond has been given little attention as a determinant of transmembrane helix association. Stimulated by recent calculations suggesting that such bonds can be much stronger than has been supposed, we have analyzed 11 known membrane protein structures and found that apparent carbon alpha hydrogen bonds cluster frequently at glycine-, serine-, and threonine-rich packing interfaces between transmembrane helices. Parallel right-handed helix-helix interactions appear to favor Calpha---H...O bond formation. In particular, Calpha --H...O interactions are frequent between helices having the structural motif of the glycophorin A dimer and the GxxxG pair. We suggest that Calpha---H...O hydrogen bonds are important determinants of stability and, depending on packing, specificity in membrane protein folding. PMID- 11481471 TI - An essential role of the JAK-STAT pathway in ischemic preconditioning. AB - The goal of this study was to determine the role of the Janus tyrosine kinase (JAK)-signal transducers and activators of transcription (STAT) pathway in the late phase of ischemic preconditioning (PC). A total of 230 mice were used. At 5 min after ischemic PC (induced with six cycles of 4-min coronary occlusion/4-min reperfusion), immunoprecipitation with anti-phosphotyrosine (anti-pTyr) antibodies followed by immunoblotting with anti-JAK antibodies revealed increased tyrosine phosphorylation of JAK1 (+257 +/- 53%) and JAK2 (+238 +/- 35%), indicating rapid activation of these two kinases. Similar results were obtained by immunoblotting with anti-pTyr-JAK1 and anti-pTyr-JAK2 antibodies. Western analysis with anti-pTyr-STAT antibodies demonstrated a marked increase in nuclear pTyr-STAT1 (+301 +/- 61%) and pTyr-STAT3 (+253 +/- 60%) 30 min after ischemic PC, which was associated with redistribution of STAT1 and STAT3 from the cytosolic to the nuclear fraction and with an increase in STAT1 and STAT3 gamma-IFN activation site DNA-binding activity (+606 +/- 64%), indicating activation of STAT1 and STAT3. No nuclear translocation or tyrosine phosphorylation of STAT2, STAT4, STAT5A, STAT5B, or STAT6 was observed. Pretreatment with the JAK inhibitor AG-490 20 min before the six occlusion/reperfusion cycles blocked the enhanced tyrosine phosphorylation of JAK1 and JAK2 and the increased tyrosine phosphorylation, nuclear translocation, and enhanced DNA-binding activity of STAT1 and STAT3. The same dose of AG-490 abrogated the protection against myocardial infarction and the concomitant up-regulation of inducible NO synthase (iNOS) protein and activity observed 24 h after ischemic PC. Taken together, these results demonstrate that ischemic PC induces isoform-selective activation of JAK1, JAK2, STAT1, and STAT3, and that ablation of this response impedes the up-regulation of iNOS and the concurrent acquisition of ischemic tolerance. This study demonstrates that the JAK-STAT pathway plays an essential role in the development of late PC. The results reveal a signaling mechanism that underlies the transcriptional up-regulation of the cardiac iNOS gene and the adaptation of the heart to ischemic stress. PMID- 11481473 TI - Physical basis of two-tone interference in hearing. AB - The cochlea uses active amplification to capture faint sounds. It has been proposed that the amplifier comprises a set of self-tuned critical oscillators: each hair cell contains a force-generating dynamical system that is maintained at the threshold of an oscillatory instability, or Hopf bifurcation. While the active response to a pure tone provides frequency selectivity, exquisite sensitivity, and wide dynamic range, its intrinsic nonlinearity causes tones of different frequency to interfere with one another in the cochlea. Here we determine the response to two tones, which provides a framework for understanding how the ear processes the more complex sounds of speech and music. Our calculations of two-tone suppression and the spectrum of distortion products generated by a critical oscillator accord with experimental observations of basilar membrane motion and the nervous response. We discuss how the response of a set of self-tuned oscillators, covering a range of characteristic frequencies, represents the structure of a complex sound. The frequency components of the stimulus can be inferred from the timing of neural spikes elicited by the vibrating hair cells. Passive prefiltering by the basilar membrane improves pitch discrimination by reducing interference between tones. Our analysis provides a general framework for examining the relation between the physical nature of the peripheral detection apparatus and psychophysical phenomena such as the sensation of dissonance and auditory illusions. PMID- 11481474 TI - ATP transduces signals from ASGM1, a glycolipid that functions as a bacterial receptor. AB - The flagella of the Gram-negative bacterium Pseudomonas aeruginosa serve not only for motility but also to bind bacteria to the host cell glycolipid asialoGM1 (ASGM1) through the protein flagellin. This interaction triggers defensive responses in host cells. How this response occurs is unclear because ASGM1 lacks transmembrane and cytoplasmic domains and there is little information about the downstream effectors that connect ASGM1 ligation to the initiation of host defense responses. Here, we show that ASGM1 ligation promotes ATP release from the host cell, followed by autocrine activation of a nucleotide receptor. This response links ASGM1 to cytoplasmic signaling molecules and results in activation of phospholipase C, Ca(2+) mobilization, phosphorylation of a mitogen-activated protein kinase (Erk 1/2), and activation of mucin transcription. These results indicate that bacterial interaction with host cells can trigger autocrine nucleotide signaling and suggest that agents affecting nucleotide receptors may modulate host responses to bacteria. PMID- 11481475 TI - ATR inhibition selectively sensitizes G1 checkpoint-deficient cells to lethal premature chromatin condensation. AB - Premature chromatin condensation (PCC) is a hallmark of mammalian cells that begin mitosis before completing DNA replication. This lethal event is prevented by a highly conserved checkpoint involving an unknown, caffeine-sensitive mediator. Here, we have examined the possible involvement of the caffeine sensitive ATM and ATR protein kinases in this checkpoint. We show that caffeine's ability to inhibit ATR (but not ATM) causes PCC, that ATR (but not ATM) prevents PCC, and that ATR prevents PCC via Chk-1 regulation. Moreover, mimicking cancer cell phenotypes by disrupting normal G(1) checkpoints sensitizes cells to PCC by ATR inhibition plus low-dose DNA damage. Notably, loss of p53 function potently sensitizes cells to PCC caused by ATR inhibition by a small molecule. We present a molecular model for how ATR prevents PCC and suggest that ATR represents an attractive therapeutic target for selectively killing cancer cells by premature chromatin condensation. PMID- 11481476 TI - The sorbin homology domain: a motif for the targeting of proteins to lipid rafts. AB - On phosphorylation of Cbl, the c-Cbl-associated protein (CAP)/Cbl complex dissociates from the insulin receptor and translocates to a lipid raft membrane fraction to form a ternary complex with flotillin. Deletion analyses of the CAP gene identified a 115-aa region responsible for flotillin binding. This region is homologous to the peptide sorbin and is referred to as the sorbin homology (SoHo) domain. This domain is present in two other proteins, vinexin and ArgBP2. Vinexin also interacted with flotillin, and deletion of its SoHo domain similarly blocked flotillin binding. The overexpression of a CAP mutant in which the SoHo domain had been deleted (CAPDeltaSoHo) prevented the translocation of Cbl to lipid rafts and subsequently blocked the recruitment of CrkII and C3G. Moreover, overexpression of CAPDeltaSoHo prevented the stimulation of glucose transport and GLUT4 translocation by insulin. These results suggest a mechanism for localization of signaling proteins to the lipid raft that mediates the compartmentalization of crucial signal transduction pathways. PMID- 11481477 TI - A mutant plasma membrane ATPase, Pma1-10, is defective in stability at the yeast cell surface. AB - Pma1 is a plasma membrane H(+)-ATPase whose activity at the cell surface is essential for cell viability. In this paper we describe a temperature-sensitive pma1 allele, pma1-10 (with two point mutations in the first cytoplasmic loop of Pma1), in which the newly synthesized mutant protein fails to remain stable at the cell surface at 37 degrees C. Instead, Pma1-10 appears to undergo internalization for vacuolar degradation in a manner dependent on End4, Vps27, Doa4, and Pep4. By contrast with wild-type Pma1, mutant Pma1-10 is hypophosphorylated and fails to associate with a Triton-insoluble fraction at 37 degrees C, suggesting failure to enter lipid rafts. Kinetic analysis reveals that, at the permissive temperature, newly synthesized Pma1-10 acquires Triton insolubility before becoming stabilized. We suggest that phosphorylation and lipid raft association may play important roles in maintaining protein stability at the plasma membrane. PMID- 11481478 TI - alpha-Synuclein occurs in lipid-rich high molecular weight complexes, binds fatty acids, and shows homology to the fatty acid-binding proteins. AB - alpha-Synuclein (alphaS) is a 140-residue neuronal protein that forms insoluble cytoplasmic aggregates in Parkinson's disease (PD) and several other neurodegenerative disorders. Two missense mutations (A53T and A30P) are linked to rare forms of familial PD. The normal function of alphaS is unknown, and cultured cell systems that model its modification from soluble monomers to aggregated forms have not been reported. Through a systematic centrifugal fractionation of mesencephalic neuronal cell lines and transgenic mouse brains expressing wild type or A53T human alphaS, we observed unusual, previously unrecognized species of alphaS that migrate well above the 17-kDa monomeric form in denaturing gels. Incubation at 65 degrees C of high-speed cytosols from cells or brains revealed a modified alphaS species migrating at approximately 36 kDa and an extensive higher molecular mass alphaS-reactive smear. Extraction of the cytosols with chloroform/methanol or with a resin (Lipidex 1000) that binds fatty acids resulted in a similar pattern of higher molecular mass alphaS forms. On the basis of this effect of delipidation, we reexamined the primary structure of alphaS and detected a motif at the N and C termini that is homologous to a fatty acid binding protein signature. In accord, we found that purified human alphaS binds oleic acid, with an apparent K(d) of 12.5 microM. We also observed an enhanced association of A53T alphaS with microsomal membranes in both mesencephalic cells and transgenic mouse brains. We conclude that alphaS has biochemical properties and a structural motif that suggest it is a novel member of the fatty acid binding protein family and may thus transport fatty acids between the aqueous and membrane phospholipid compartments of the neuronal cytoplasm. PMID- 11481479 TI - Conditional epidermal expression of TGFbeta 1 blocks neonatal lethality but causes a reversible hyperplasia and alopecia. AB - To study the role of transforming growth factor type beta1 (TGFbeta1) in epidermal growth control and disease, we have generated a conditional expression system by using the bovine keratin 5 promoter to drive expression of the tetracycline-regulated transactivators tTA and rTA, and a constitutively active mutant of TGFbeta1 linked to the tetO target sequence for the transactivator. This model allows for induction or suppression of exogenous TGFbeta1 with oral doxycycline. Maximal expression of TGFbeta1 during gestation caused embryonic lethality, whereas partial suppression allowed full-term development with neonatal lethality characterized by runting, epidermal hypoproliferation, and blocked hair follicle growth. With complete suppression, phenotypically normal double transgenic (DT) mice were born. Acute induction of TGFbeta1 in the epidermis of adult mice inhibited basal and follicular keratinocyte proliferation and reentry of telogen hair follicles into anagen. However, chronic expression of TGFbeta1 in adult DTs caused severe alopecia characterized by epidermal and follicular hyperproliferation, apoptosis, as well as dermal fibrosis and inflammation. Readministration of doxycycline to tTA DT mice caused hair regrowth within 14 days. The mRNA and protein for Smad7, an inhibitor of TGFbeta signaling, were up-regulated in the epidermis and hair follicles of alopecic skin and rapidly induced in rTA mice in parallel with the TGFbeta1 transgene, suggesting that the hyperproliferative phenotype may result in part from development of a sustained negative feedback loop. Thus, this conditional expression system provides an important model for understanding the role of TGFbeta1 during development, in normal skin biology, and in disease. PMID- 11481480 TI - Specific repression of beta-globin promoter activity by nuclear ferritin. AB - Developmental hemoglobin switching involves sequential globin gene activations and repressions that are incompletely understood. Earlier observations, described herein, led us to hypothesize that nuclear ferritin is a repressor of the adult beta-globin gene in embryonic erythroid cells. Our data show that a ferritin family protein in K562 cell nuclear extracts binds specifically to a highly conserved CAGTGC motif in the beta-globin promoter at -153 to -148 bp from the cap site, and mutation of the CAGTGC motif reduces binding 20-fold in competition gel-shift assays. Purified human ferritin that is enriched in ferritin-H chains also binds the CAGTGC promoter segment. Expression clones of ferritin-H markedly repress beta-globin promoter-driven reporter gene expression in cotransfected CV 1 cells in which the beta-promoter has been stimulated with the transcription activator erythroid Kruppel-like factor (EKLF). We have constructed chloramphenicol acetyltransferase reporter plasmids containing either a wild-type or mutant beta-globin promoter for the -150 CAGTGC motif and have compared the constructs for susceptibility to repression by ferritin-H in cotransfection assays. We find that stimulation by cotransfected EKLF is retained with the mutant promoter, whereas repression by ferritin-H is lost. Thus, mutation of the 150 CAGTGC motif not only markedly reduces in vitro binding of nuclear ferritin but also abrogates the ability of expressed ferritin-H to repress this promoter in our cell transfection assay, providing a strong link between DNA binding and function, and strong support for our proposal that nuclear ferritin-H is a repressor of the human beta-globin gene. Such a repressor could be helpful in treating sickle cell and other genetic diseases. PMID- 11481481 TI - How cuckoldry can decrease the opportunity for sexual selection: data and theory from a genetic parentage analysis of the sand goby, Pomatoschistus minutus. AB - Alternative mating strategies are common in nature and are generally thought to increase the intensity of sexual selection. However, cuckoldry can theoretically decrease the opportunity for sexual selection, particularly in highly polygamous species. We address here the influence of sneaking (fertilization thievery) on the opportunity for sexual selection in the sand goby Pomatoschistus minutus, a marine fish species in which males build and defend nests. Our microsatellite based analysis of the mating system in a natural sand goby population shows high rates of sneaking and multiple mating by males. Sneaker males had fertilized eggs in approximately 50% of the assayed nests, and multiple sneakers sometimes fertilized eggs from a single female. Successful males had received eggs from 2 to 6 females per nest (mean = 3.4). We developed a simple mathematical model showing that sneaking in this polygynous sand goby population almost certainly decreases the opportunity for sexual selection, an outcome that contrasts with the usual effects of cuckoldry in socially monogamous animals. These results highlight a more complex and interesting relationship between cuckoldry rates and the intensity of sexual selection than previously assumed in much of the literature on animal mating systems. PMID- 11481482 TI - Efficient chromosomal transposition of a Tc1/mariner- like transposon Sleeping Beauty in mice. AB - The presence of mouse embryonic stem (ES) cells makes the mouse a powerful model organism for reverse genetics, gene function study through mutagenesis of specific genes. In contrast, forward genetics, identification of mutated genes responsible for specific phenotypes, has an advantage to uncover novel pathways and unknown genes because no a priori assumptions are made about the mutated genes. However, it has been hampered in mice because of the lack of a system in which a large-scale mutagenesis and subsequent isolation of mutated genes can be performed efficiently. Here, we demonstrate the efficient chromosomal transposition of a Tc1/mariner-like transposon, Sleeping Beauty, in mice. This system allows germ-line mutagenesis in vivo and will facilitate certain aspects of phenotype-driven genetic screening in mice. PMID- 11481483 TI - Comparative expressed sequence hybridization to chromosomes for tumor classification and identification of genomic regions of differential gene expression. AB - Altered expression of genes can have phenotypic consequences in cancer development and treatment, developmental abnormalities, and differentiation processes. Here we describe a rapid approach, termed comparative expressed sequence hybridization (CESH), which gives a genome-wide view of relative expression patterns within tissues according to chromosomal location. No prior knowledge of genes or cloning is required, and minimal amounts of tissue can be used. Expression profiles are achieved in a manner similar to the identification of chromosomal imbalances by comparative genomic hybridization analysis. The approach is demonstrated to indicate a chromosomal region that harbors overexpressed genes that may be associated with a drug-resistant phenotype. In addition, known and new regions of differential gene expression in both normal tissues and tumor samples from the soft tissue sarcoma group of rhabdomyosarcoma (RMS) are indicated. These regions included 2p24; overexpression of MYCN at 2p24 was confirmed by quantitative reverse transcription-PCR for all of the alveolar RMS cases and did not necessarily correspond to genomic amplification. Evidence including region specific microarray analysis indicated that overexpression of several genes from a region may be required for detection by CESH. This evidence is consistent with clusters of functionally related genes and mechanisms that affect the expression of a number of genes at a particular genomic location. The distinctive CESH profiles demonstrated in different subtypes of RMS show potential for tumor classification. PMID- 11481484 TI - Growth inhibition and DNA damage induced by Cre recombinase in mammalian cells. AB - The use of Cre/loxP recombination in mammalian cells has expanded rapidly. We describe here that Cre expression in cultured mammalian cells may result in a markedly reduced proliferation and that this effect is dependent on the endonuclease activity of Cre. Chromosome analysis after Cre expression revealed numerous chromosomal aberrations and an increased number of sister chromatid exchanges. Titration experiments in mouse embryo fibroblasts with a ligand regulatable Cre-ER(T) show that toxicity is dependent on the level of Cre activity. Prolonged, low levels of Cre activity permit recombination without concomitant toxicity. This urges for a careful titration of Cre activity in conditional gene modification in mammalian cells. PMID- 11481485 TI - PNA interference mapping demonstrates functional domains in the noncoding RNA Xist. AB - The noncoding RNA Xist has been shown to be essential for X-chromosome inactivation and to coat the inactive X-chromosome (Xi). Thus, an important question in understanding the formation of Xi is whether the binding reaction of Xist is necessary for X-chromosome inactivation. In this article, we demonstrate the failure of X-chromosome silencing if the association of Xist with the X chromosome is inhibited. The chromatin-binding region was functionally mapped and evaluated by using an approach for studying noncoding RNA function in living cells that we call peptide nucleic acid (PNA) interference mapping. In the reported experiments, a single 19-bp antisense cell-permeating PNA targeted against a particular region of Xist RNA caused the disruption of the Xi. The association of the Xi with macro-histone H2A is also disturbed by PNA interference mapping. PMID- 11481486 TI - A role of PDGFRalpha in basal cell carcinoma proliferation. AB - Activation of the hedgehog pathway, through the loss of patched (PTC) or the activation of smoothened (SMO), occurs frequently in basal cell carcinoma (BCC), the most common human cancer. However, the molecular basis of this neoplastic effect is not understood. The downstream molecule Gli1 is known to mediate the biological effect of the pathway and is itself up-regulated in all BCCs. Gli1 can drive the production of BCCs in the mouse when overexpressed in the epidermis. Here we show that Gli1 can activate platelet-derived growth factor receptor alpha (PDGFRalpha) in C3H10T(1/2) cells. Functional up-regulation of PDGFRalpha by Gli1 is accompanied by activation of the ras-ERK pathway, a pathway associated with cell proliferation. The relevance of this mechanism in vivo is supported by a high level expression of PDGFRalpha in BCCs of mice and humans. In the murine BCC cell line ASZ001, in which both copies of the PTC gene are inactivated, DNA synthesis and cell proliferation can be slowed by re-expression of PTC, which down-regulates PDGFRalpha expression, or by downstream inhibition of PDGFRalpha with neutralizing antibodies. Therefore, we conclude that increased expression of PDGFRalpha may be an important mechanism by which mutations in the hedgehog pathway cause BCCs. PMID- 11481487 TI - An HIV-1 transgenic rat that develops HIV-related pathology and immunologic dysfunction. AB - We report, to our knowledge, the first HIV type 1 (HIV-1) transgenic (Tg) rat. Expression of the transgene, consisting of an HIV-1 provirus with a functional deletion of gag and pol, is regulated by the viral long terminal repeat. Spliced and unspliced viral transcripts were expressed in lymph nodes, thymus, liver, kidney, and spleen, suggesting that Tat and Rev are functional. Viral proteins were identified in spleen tissue sections by immunohistochemistry and gp120 was present in splenic macrophages, T and B cells, and in serum. Clinical signs included wasting, mild to severe skin lesions, opaque cataracts, neurological signs, and respiratory difficulty. Histopathology included a selective loss of splenocytes within the periarterial lymphoid sheath, increased apoptosis of endothelial cells and splenocytes, follicular hyperplasia of the spleen, lymphocyte depletion of mesenteric lymph nodes, interstitial pneumonia, psoriatic skin lesions, and neurological, cardiac, and renal pathologies. Immunologically, delayed-type hypersensitivity response to keyhole limpet hemocyanin was diminished. By contrast, Ab titers and proliferative response to recall antigen (keyhole limpet hemocyanin) were normal. The HIV-1 Tg rat thus has many similarities to humans infected with HIV-1 in expression of viral genes, immune response alterations, and pathologies resulting from infection. The HIV-1 Tg rat may provide a valuable model for some of the pathogenic manifestations of chronic HIV-1 diseases and could be useful in testing therapeutic regimens targeted to stages of viral replication subsequent to proviral integration. PMID- 11481488 TI - Imaging transcriptional regulation of p53-dependent genes with positron emission tomography in vivo. AB - A noninvasive method for molecular imaging of the activity of different signal transduction pathways and the expression of different genes in vivo would be of considerable value. It would aid in understanding the role specific genes and signal transduction pathways have in various diseases, and could elucidate temporal dynamics and regulation at different stages of disease and during various therapeutic interventions. We developed and assessed a method for monitoring the transcriptional activation of endogenous genes by positron emission tomography (PET) imaging. The HSV1-tk/GFP (TKGFP) dual reporter gene was used to monitor transcriptional activation of p53-dependent genes. A retrovirus bearing the Cis-p53/TKGFP reporter system was constructed in which the TKGFP reporter gene was placed under control of an artificial cis-acting p53-specific enhancer. U87 glioma and SaOS-2 osteosarcoma cells were transduced with this retrovirus and used to establish xenografts in rats. We demonstrated that DNA damage-induced up-regulation of p53 transcriptional activity correlated with the expression of p53-dependent downstream genes, such as p21, in U87 (wild-type p53), but not in SaOS-2 osteosarcoma (p53 -/-) cells. We showed that PET, with [(124)I]FIAU (2'-fluoro-2'-deoxy-1-beta-d-arabinofuranosyl-5-[(124)I]iodouracil) and the Cis-p53TKGFP reporter system, is sufficiently sensitive to image the transcriptional regulation of genes in the p53 signal transduction pathway. These imaging results were confirmed by independent measurements of p53 activity and the expression levels of downstream genes (e.g., p21) by using conventional molecular-biological assays. PET imaging of p53 transcriptional activity in tumor xenografts by using the Cis-p53TKGFP reporter system may be useful in assessing novel therapeutic approaches. PMID- 11481489 TI - Placental defects and embryonic lethality in mice lacking suppressor of cytokine signaling 3. AB - Mice lacking suppressor of cytokine signaling 3 (SOCS3) exhibited embryonic lethality with death occurring between days 11 and 13 of gestation. At this stage, SOCS3(-/-) embryos were slightly smaller than wild type but appeared otherwise normal, and histological analysis failed to detect any anatomical abnormalities responsible for the lethal phenotype. Rather, in all SOCS3(-/-) embryos examined, defects were evident in placental development that would account for their developmental arrest and death. The placental spongiotrophoblast layer was significantly reduced and accompanied by increased numbers of giant trophoblast cells. Delayed branching of the chorioallantois was evident, and, although embryonic blood vessels were present in the labyrinthine layer of SOCS3(-/-) placentas, the network of embryonic vessels and maternal sinuses was poorly developed. Yolk sac erythropoiesis was normal, and, although the SOCS3(-/-) fetal liver was small at day 12.5 of gestation (E12.5), normal frequencies of erythroblasts and hematopoietic progenitor cells, including blast forming unit-erythroid (BFU-E) and, colony forming unit-erythroid (CFU-E) were present at both E11.5 and E12.5. Colony formation for both BFU-E and CFU-E from SOCS3(-/-) mice displayed wild-type quantitative responsiveness to erythropoietin (EPO), in the presence or absence of IL-3 or stem cell factor (SCF). These data suggest that SOCS3 is required for placental development but dispensable for normal hematopoiesis in the mouse embryo. PMID- 11481490 TI - An inherited p53 mutation that contributes in a tissue-specific manner to pediatric adrenal cortical carcinoma. AB - The incidence of pediatric adrenal cortical carcinoma (ACC) in southern Brazil is 10-15 times higher than that of pediatric ACC worldwide. Because childhood ACC is associated with Li-Fraumeni syndrome, we examined the cancer history and p53 status of 36 Brazilian patients and their families. Remarkably, 35 of 36 patients had an identical germ-line point mutation of p53 encoding an R337H amino acid substitution. Differences within intragenic polymorphic markers demonstrated that at least some mutant alleles arose independently, thus eliminating a founder effect. In tumor cells, the wild-type allele was deleted, and mutant p53 protein accumulated within the nuclei. Although these features are consistent with Li Fraumeni syndrome-associated adrenal tumors, there was no history of increased cancer incidence among family members. Therefore, this inherited R337H p53 mutation represents a low-penetrance p53 allele that contributes in a tissue specific manner to the development of pediatric ACC. PMID- 11481491 TI - Gene program for cardiac cell survival induced by transient ischemia in conscious pigs. AB - Therapy for ischemic heart disease has been directed traditionally at limiting cell necrosis. We determined by genome profiling whether ischemic myocardium can trigger a genetic program promoting cardiac cell survival, which would be a novel and potentially equally important mechanism of salvage. Although cardiac genomics is usually performed in rodents, we used a swine model of ischemia/reperfusion followed by ventricular dysfunction (stunning), which more closely resembles clinical conditions. Gene expression profiles were compared by subtractive hybridization between ischemic and normal tissue of the same hearts. About one third (23/74) of the nuclear-encoded genes that were up-regulated in ischemic myocardium participate in survival mechanisms (inhibition of apoptosis, cytoprotection, cell growth, and stimulation of translation). The specificity of this response was confirmed by Northern blot and quantitative PCR. Unexpectedly, this program also included genes not previously described in cardiomyocytes. Up regulation of survival genes was more profound in subendocardium over subepicardium, reflecting that this response in stunned myocardium was proportional to the severity of the ischemic insult. Thus, in a swine model that recapitulates human heart disease, nonlethal ischemia activates a genomic program of cell survival that relates to the time course of myocardial stunning and differs transmurally in relation to ischemic stress, which induced the stunning. Understanding the genes up-regulated during myocardial stunning, including those not previously described in the heart, and developing strategies that activate this program may open new avenues for therapy in ischemic heart disease. PMID- 11481492 TI - A genomic island in Pseudomonas aeruginosa carries the determinants of flagellin glycosylation. AB - Protein glycosylation has been long recognized as an important posttranslational modification process in eukaryotic cells. Glycoproteins, predominantly secreted or surface localized, have also been identified in bacteria. We have identified a cluster of 14 genes, encoding the determinants of the flagellin glycosylation machinery in Pseudomonas aeruginosa PAK, which we called the flagellin glycosylation island. Flagellin glycosylation can be detected only in bacteria expressing the a-type flagellin sequence variants, and the survey of 30 P. aeruginosa isolates revealed coinheritance of the a-type flagellin genes with at least one of the flagellin glycosylation island genes. Expression of the b-type flagellin in PAK, an a-type strain carrying the glycosylation island, did not lead to glycosylation of the b-type flagellin of PAO1, suggesting that flagellins expressed by b-type bacteria not only lack the glycosylation island, they cannot serve as substrates for glycosylation. Providing the entire glycosylation island of PAK, including its a-type flagellin in a flagellin mutant of a b-type strain, results in glycosylation of the heterologous flagellin. These results suggest that some or all of the 14 genes on the glycosylation island are the genes that are missing from strain PAO1 to allow glycosylation of an appropriate flagellin. Inactivation of either one of the two flanking genes present on this island abolished flagellin glycosylation. Based on the limited homologies of these gene products with enzymes involved in glycosylation, we propose that the island encodes similar proteins involved in synthesis, activation, or polymerization of sugars that are necessary for flagellin glycosylation. PMID- 11481493 TI - Globin-coupled sensors: a class of heme-containing sensors in Archaea and Bacteria. AB - The recently discovered prokaryotic signal transducer HemAT, which has been described in both Archaea and Bacteria, mediates aerotactic responses. The N terminal regions of HemAT from the archaeon Halobacterium salinarum (HemAT-Hs) and from the Gram-positive bacterium Bacillus subtilis (HemAT-Bs) contain a myoglobin-like motif, display characteristic heme-protein absorption spectra, and bind oxygen reversibly. Recombinant HemAT-Hs and HemAT-Bs shorter than 195 and 176 residues, respectively, do not bind heme effectively. Sequence homology comparisons and three-dimensional modeling predict that His-123 is the proximal heme-binding residue in HemAT from both species. The work described here used site-specific mutagenesis and spectroscopy to confirm this prediction, thereby providing direct evidence for a functional domain of prokaryotic signal transducers that bind heme in a globin fold. We postulate that this domain is part of a globin-coupled sensor (GCS) motif that exists as a two-domain transducer having no similarity to the PER-ARNT-SIM (PAS)-domain superfamily transducers. Using the GCS motif, we have identified several two-domain sensors in a variety of prokaryotes. We have cloned, expressed, and purified two potential globin-coupled sensors and performed spectral analysis on them. Both bind heme and show myoglobin-like spectra. This observation suggests that the general function of GCS-type transducers is to bind diatomic oxygen and perhaps other gaseous ligands, and to transmit a conformational signal through a linked signaling domain. PMID- 11481494 TI - Functional expression and characterization of a Drosophila odorant receptor in a heterologous cell system. AB - Odorant receptors (ORs) constitute the molecular basis for the detection of volatile odorous molecules and the perception of smell. Our understanding of chemical senses has been greatly expanded by the discovery of the OR gene families in vertebrates and in the nematode Caenorhabditis elegans. Recently, candidate Drosophila OR genes have been identified. The putative ORs do not possess any primary sequence identity with known vertebrate or C. elegans receptors, but belong to the family of G protein-coupled receptors according to their predicted seven transmembrane topology. To prove olfactory function of these proteins, we expressed a member of the putative Drosophila OR gene family, Or43a, in Xenopus laevis oocytes. Using two-electrode voltage-clamp recording we identified four odors (cyclohexanone, cyclohexanol, benzaldehyde, and benzyl alcohol) that activated the receptor at low micromolar concentration and structurally related substances that did not. This report shows the function and specificity of a member of the recently identified family of Drosophila ORs expressed in a heterologous system. PMID- 11481495 TI - Functional analysis of an olfactory receptor in Drosophila melanogaster. AB - Fifty nine candidate olfactory receptor (Or) genes have recently been identified in Drosophila melanogaster, one of which is Or43a. In wild-type flies, Or43a is expressed at the distal edge of the third antennal segment in about 15 Or neurons. To identify ligands for the receptor we used the Gal4/UAS system to misexpress Or43a in the third antennal segment. Or43a mRNA expression in the antenna of transformed and wild-type flies was visualized by in situ hybridization with a digoxigenin-labeled probe. Electroantennogram recordings from transformed and wild-type flies were used to identify cyclohexanol, cyclohexanone, benzaldehyde, and benzyl alcohol as ligands for the Or43a. This in vivo analysis reveals functional properties of one member of the recently isolated Or family in Drosophila and will provide further insight into our understanding of olfactory coding. PMID- 11481496 TI - Distinct mechanisms of cell cycle arrest control the decision between differentiation and senescence in human neuroblastoma cells. AB - Retinoic acid (RA) induces cell cycle arrest and differentiation of human neuroblastoma (NB) cells. Typically, NB cells differentiate along the neuronal lineage, but quiescent, "flat" cell types frequently have been described after treatment with differentiating agents. Two indistinguishable subclones of the cell line SK-N-SH, SK-N-SH-N (SH-N) and SK-N-SH-F (SH-F), display dramatically different responses to RA. In SH-N, RA induces neuronal differentiation, but in SH-F it transforms the small neuroblastic cells into large, flattened, epithelium like cells. Here we analyze the mechanistic basis for the different effects of RA in the two NB subclones. First, we show that the flattened RA-treated SH-F expresses markers of cells undergoing replicative senescence. Inhibition of DNA synthesis by RA is significantly more rapid in SH-F than in SH-N. SH-F, which expresses basal amounts of p16(INK4A), responds to RA with elevation of p18(INK4C), marked down-regulation of cyclin D1, and swift inhibition of cyclin D dependent kinases (cdks). Conversely, after addition of RA, SH-N retains cell cycling due to high expression of cyclin D1, the absence of Ink4 inhibitors, and accumulation of p21(Cip1). These changes result in sustained cdk activity. Accordingly, overexpression of p21(Cip1) but not p16(INK4A) induces neuronal differentiation of untreated NB cells. We propose that rapid inhibition of cdks by RA in NB leads to early cell cycle arrest, prevents neuronal differentiation, and results in a senescence-like state. PMID- 11481497 TI - Effects of forced diving on the spleen and hepatic sinus in northern elephant seal pups. AB - In phocid seals, an increase in hematocrit (Hct) accompanies diving and periods of apnea. The variability of phocid Hct suggests that the total red cell mass is not always in circulation, leading researchers to speculate on the means of blood volume partitioning. The histology and disproportionate size of the phocid spleen implicates it as the likely site for RBC storage. We used magnetic resonance imaging on Northern elephant seals to demonstrate a rapid contraction of the spleen and a simultaneous filling of the hepatic sinus during forced dives (P < 0.0001, R(2) = 0.97). The resulting images are clear evidence demonstrating a functional relationship between the spleen and hepatic sinus. The transfer of blood from the spleen to the sinus provides an explanation for the disparity between the timing of diving-induced splenic contraction ( approximately 1-3 min) and the occurrence of peak Hct (15-25 min). Facial immersion was accompanied by an immediate and profound splenic contraction, with no further significant decrease in splenic volume after min 2 (Tukey-Kramer HSD, P = 0.05). At the conclusion of the dive, the spleen had contracted to 16% of its predive volume (mean resting splenic volume = 3,141 ml +/- 68.01 ml; 3.54% of body mass). In the postdive period, the spleen required 18-22 min to achieve resting volume, indicating that this species may not have sufficient time to refill the spleen when routinely diving at sea, which is virtually continuous with interdive surface intervals between 1 and 3 min. PMID- 11481498 TI - Multiple transcription-factor genes are early targets of phytochrome A signaling. AB - The phytochrome family of sensory photoreceptors directs adaptational changes in gene expression in response to environmental light signals. Using oligonucleotide microarrays to measure expression profiles in wild-type and phytochrome A (phyA) null-mutant Arabidopsis seedlings, we have shown that 10% of the genes represented on the array are regulated by phyA in response to a continuous far red light signal. Strikingly, 44% of the genes responding to the signal within 1 h are predicted to encode multiple classes of transcriptional regulators. Together with previous data, this observation suggests that phyA may regulate seedling photomorphogenesis by direct targeting of light signals to the promoters of genes encoding a master set of diverse transcriptional regulators, responsible in turn for orchestrating the expression of multiple downstream target genes in various branches of a phyA-regulated transcriptional network. PMID- 11481499 TI - Transpiration, a prerequisite for long-distance transport of minerals in plants? AB - The major "benefit" alleged to accrue from transpiration (the evaporative loss of water from plant surfaces) is that it is essential for the long-distance transport of mineral ions, but the possible interrelation between these two processes has rarely been tested. Transpiration was experimentally dissociated from mineral supply by growing sunflowers (Helianthus anuus) in hydroculture and providing mineral nutrients only during the nights. These plants grew as well as a control group that received nutrients only during the day and transpired 12-15 times more water during the exposure period. It thus appears that convective water transport in the xylem, brought about by root pressure and the resultant guttation, "growth water," and Munch's phloem counterflow is in itself sufficient for long-distance mineral supply and that transpiration is not required for this function. PMID- 11481500 TI - A fatty acid desaturase modulates the activation of defense signaling pathways in plants. AB - Salicylic acid (SA) plays an important role in activating various plant defense responses, including expression of the pathogenesis-related (PR) genes and systemic acquired resistance. A critical positive regulator of the SA signaling pathway in Arabidopsis is encoded by the NPR1 gene. However, there is growing evidence that NPR1-independent pathways can also activate PR expression and disease resistance. To elucidate the components associated with NPR1-independent defense signaling, we isolated a suppressor of the npr1-5 allele, designated ssi2. The recessive ssi2 mutation confers constitutive PR gene expression, spontaneous lesion formation, and enhanced resistance to Peronospora parasitica. In contrast, a subset of defense responses regulated by the jasmonic acid (JA) signaling pathway, including expression of the defensin gene PDF1.2 and resistance to Botrytis cinerea, is impaired in ssi2 plants. With the use of a map based approach, the SSI2 gene was cloned and shown to encode a stearoyl-ACP desaturase (S-ACP DES). S-ACP DES is an archetypical member of a family of soluble fatty acid (FA) desaturases; these enzymes play an important role in regulating the overall level of desaturated FAs in the cell. The activity of mutant S-ACP DES enzyme was reduced 10-fold, resulting in elevation of the 18:0 FA content in ssi2 plants. Because reduced S-ACP DES activity leads to the induction of certain defense responses and the inhibition of others, we propose that a FA-derived signal modulates crosstalk between different defense signaling pathways. PMID- 11481501 TI - The Y chromosome in the liverwort Marchantia polymorpha has accumulated unique repeat sequences harboring a male-specific gene. AB - The haploid liverwort Marchantia polymorpha has heteromorphic sex chromosomes, an X chromosome in the female and a Y chromosome in the male. We here report on the repetitive structure of the liverwort Y chromosome through the analysis of male specific P1-derived artificial chromosome (PAC) clones, pMM4G7 and pMM23-130F12. Several chromosome-specific sequence elements of approximately 70 to 400 nt are combined into larger arrangements, which in turn are assembled into extensive Y chromosome-specific stretches. These repeat sequences contribute 2-3 Mb to the Y chromosome based on the observations of three different approaches: fluorescence in situ hybridization, dot blot hybridization, and the frequency of clones containing the repeat sequences in the genomic library. A novel Y chromosome specific gene family was found embedded among these repeat sequences. This gene family encodes a putative protein with a RING finger motif and is expressed specifically in male sexual organs. To our knowledge, there have been no other reports for an active Y chromosome-specific gene in plants. The chromosome specific repeat sequences possibly contribute to determining the identity of the Y chromosome in M. polymorpha as well as to maintaining genes required for male functions, as in mammals such as human. PMID- 11481502 TI - Reproductive constraints and the evolution of life histories with indeterminate growth. AB - A prominent feature of comparative life histories in fish (and other indeterminate growers) is the approximate invariance across species of certain dimensionless numbers made up from reproductive and timing variables. The two best known are the age at maturity (alpha) divided by the average adult lifespan (E), and the proportion of a body mass given to reproduction per year (c) multiplied by E. This article uses evolutionary life-history theory for nongrowing populations to predict the numeric value of these numbers for fish and lizards, with several new implications for the dynamics of ontogenetic development. PMID- 11481503 TI - Change in the diameter of the radial artery transradial intervention using a 6 French system in Japanese patients. AB - We examined the change in the diameter of the radial artery after transradial intervention (TRI) using a 6 French system in 50 Japanese patients in order to investigate whether the radial artery would be preserved after TRI. In all cases, the Allen test was normal and repeated puncture was possible at the TRI site 3 months after TRI. Moreover, the appearance of new stenosis or thinning throughout the radial artery at the TRI site was not seen. The diameter of the radial artery 3 months after TRI tended to be somewhat smaller at both the proximal and distal sites; however, there was not a statistically significant change between the diameter before and 3 months after TRI. Our results show that the radial artery will be preserved after TRI using a 6 French system in selected Japanese patients. PMID- 11481504 TI - Transradial cardiac catheterization and the use of the radial artery as a bypass graft. PMID- 11481505 TI - Sidebranch occlusion in direct intracoronary stenting: predictors and results. AB - We assessed the angiographic predictors and results of major (> 1 mm) sidebranch occlusion (SBO) following direct intracoronary stenting (DS) in 86 patients. The occlusion of a sidebranch is a well-defined risk after balloon angioplasty and stenting. However, the impact of direct stenting without predilatation on the coronary flow of sidebranches emerging within the stented segment has not been studied solely. A total of 111 sidebranches were analyzed. Sidebranch type, take off angle, ostial involvement and procedural characteristics were evaluated. Nine out of 111 (8%) stent-covered sidebranches were occluded. Sidebranches with > 50% stenosis that take off within or just beyond the diseased portion of the lesion (unfavorable morphology) were the most powerful morphologic predictor of SBO (odds ratio: 8.0; 95% confidence interval: 1.5--40.8; p = 0.007). Take-off angle of the sidebranch was not found to have any effect on SBO. Post-stent dilation using high-pressure inflation (15.0 +/- 2.1 atmospheres), inflation (odds ratio: 1.1; 95% CI: 1.0--1.2; p = 0.038), and 3 times inflation (odds ratio: 4.5; 95% CI: 1.1--18.3; p = 0.023) were the procedural predictor of SBO. Of those 40 unfavorable morphologies, seven (17.5%) were occluded compared to 2.8% (2/71) of the favorable morphologies. Nine out of 9 (100%) occluded after high-pressure inflation. Three patients complained of chest pain and 1 had non-Q wave myocardial infarction attributed to SBO. These findings indicate that the incidence of SBO and complications are less than expected and has a favorable outcome in direct intracoronary stenting. PMID- 11481506 TI - Sidebranch occlusion in direct intracoronary stenting: predictors and results. PMID- 11481507 TI - Impact of pre-stent plaque debulking for chronic coronary total occlusions on restenosis reduction. AB - BACKGROUND: While stenting improves the long-term angiographic outcomes of successfully recanalized chronic coronary total occlusions (CTO), the restenosis rate still remains high. The massive plaque burden in CTO is considered to be one of the causes of in-stent restenosis. METHODS: We examined the pre-stent plaque debulking strategy with high-speed rotational atherectomy (RA) for 50 CTO (Thrombolysis in Myocardial Infarction flow grade 0; estimated occlusive duration, 3 months). Angiographic follow-up results were compared to those of 120 consecutive CTO recanalized with primary stenting in which RA could be indicated retrospectively. Angiographic restenosis was defined as diameter stenosis > 50% at 6-month follow-up. RESULTS: RA could be performed safely in all lesions without any major complications. Adjunctive ballooning and stenting could be performed without high-pressure dilatation (8.4 +/- 1.7 atmospheres). Follow up angiography was performed in 48 lesions 184 +/- 61 days after the procedure. There were no significant differences in baseline characteristics between the two groups; however, the implanted stent type was different. Quantitative coronary angiography revealed that diameter stenosis was smaller at follow-up (36.2 +/- 20.0% versus 52.2 +/- 26.7%; p = 0.0003) as well as post-procedure (7.8 +/- 11.5% versus 17.8 +/- 13.6%; p < 0.0001) compared with the control group. Angiographic restenosis was also significantly reduced (29.2% versus 52.5%; p = 0.0061). CONCLUSIONS: RA is a safe procedure for plaque debulking of CTO in selected cases. Plaque debulking of CTO facilitates subsequent stent expansion and may reduce the restenosis rate. PMID- 11481508 TI - Chronic total coronary occlusions: an interventional cardiology nemesis. PMID- 11481509 TI - Percutaneous suture closure for management of large French size arterial puncture in aortic valvuloplasty. AB - Percutaneous aortic valvuloplasty produces acute hemodynamic and clinical improvement in patients with aortic stenosis who are poor candidates for surgical valve replacement. The benefits of this procedure are mitigated by the high restenosis rates and 1--2 year mortality, in addition to significant vascular complications associated with the large arterial puncture necessary for retrograde arterial approaches. We describe the use of suture closure techniques to reduce the vascular morbidity associated with this procedure. We reviewed 31 consecutive patients who underwent percutaneous aortic valvuloplasty and suture closure with the Perclose device between April 1998 and September 2000. After diagnostic catheterization using 6--8 French (Fr) sheaths, an 8 or 10 Fr Perclose device was preloaded into the artery and the untied sutures left in place. A 12.5 or 14 Fr sheath was passed over the wire, through the sutures. The sutures were tied at the conclusion of the procedure, in conjunction with sheath removal. Twenty-seven out of 31 patients (87%) had successful suture closure of the arteriotomy and did not require prolonged bed rest, manual compression or a compression device. There were 4 failures of percutaneous suture closure, requiring conventional manual compression and bed rest for hemostasis. No patient required surgical repair of the arteriotomy, nor were there any limb complications. Compared to 39 consecutive prior patients who had their arterial puncture managed with manual compression, length of stay was shorter (2.2 days versus 5.3 days) and fewer patients received blood transfusions (0% versus 29%). Preloaded suture closure of the arterial puncture is a useful technique for achieving hemostasis after removal of the large sheaths used for percutaneous valvuloplasty, and reduces the post-procedure patient discomfort and prolonged bed rest associated with this procedure. PMID- 11481510 TI - Acute myocardial infarction in a young female. PMID- 11481511 TI - Endothelial dysfunction and progressive coronary atherosclerosis: sequential invasive studies in a patient with multiple cardiac risk factors. AB - This manuscript describes sequential angiographic, endothelial and vasoreactivity characteristics of the coronary arterial circulation in a middle-aged patient with multiple cardiac risk factors who developed hemodynamically significant coronary atherosclerosis over a 6-year period. A 56-year-old woman demonstrated marked angiographic progression of coronary atherosclerosis over time beginning with minor luminal irregularities in the setting of severe endothelial dysfunction. The association of endothelial dysfunction, ineffective cardiac risk factor management and progressive atherosclerosis is discussed. PMID- 11481512 TI - Intravascular ultrasound assessment and endoluminal stenting of a stenosis in a 50-month-old coronary polytetrafluoroethylene graft. AB - We report the assessment and management of an 81-year-old patient with a rare stenosis in a 50-month-old polytetrafluoroethylene coronary graft. We concluded that atheromatous plaque was the cause for the stenosis based on the lesion location within the graft and its characteristics by intravascular ultrasound. Differential diagnoses are discussed under consideration of the literature. The lesion was successfully treated using endoluminal stenting. PMID- 11481513 TI - Why endovascular therapy should be utilized before surgical bypass for femoropopliteal occlusive disease. PMID- 11481514 TI - Anatomy of a pseudo-controversy, or how polarization is often turf-based! PMID- 11481515 TI - Interview with William S. Weintraub, MD. [interviewed by Richard E. Shaw]. PMID- 11481516 TI - Right heart catheterization via the antecubital vein: a forgotten technique? PMID- 11481517 TI - Postirradiation pseudosclerodermatous panniculitis. AB - Pseudosclerodermatous panniculitis is an unusual variant of panniculitis that results as a complication of megavoltage radiotherapy. Four women developed this unusual entity on the anterior chest and abdominal skin after receiving megavoltage therapy for either breast carcinoma or painful bone metastases from breast carcinoma. Histopathologically, the epidermis and dermis of the involved area showed little or no evidence of radiodermatitis. The main findings were confined to the subcutaneous tissue and consisted of thickened, sclerotic septa composed of both thick and thin collagen bundles, and a lobular panniculitis characterized by lipophagic granulomas and scattered lymphocytes and plasma cells. Additionally, one of the cases showed markedly dilated vascular spaces with the appearance of lymphatics in the upper part of the dermis. Pseudosclerodermatous panniculitis after irradiation is an unusual cutaneous complication of megavoltage radiotherapy that should be distinguished from subcutaneous metastatic disease, cellulitis, or connective tissue diseases involving the subcutaneous fat. The differential diagnosis can be established on the basis of the characteristic histopathologic features of postirradiation pseudosclerodermatous panniculitis. PMID- 11481518 TI - The p75 neurotrophin receptor, relative to other Schwann cell and melanoma markers, is abundantly Expressed in spindled melanomas. AB - Seventeen cases of spindled melanomas and eleven cases of epithelioid melanomas were immunolabeled with various melanoma and Schwann cell markers. Standard melanoma markers included S100, HMB45, HMB50, tyrosinase, and Melan A. Schwann cell markers included the p75 neurotrophin receptor (p75NTR), glial fibrillary acidic protein (GFAP), and the L1 adhesion protein. The degree of immunocytochemical labeling was scored by levels of both intensity and pervasiveness. The results confirmed a distinct difference in labeling between epithelioid and spindled melanomas. The p75NTR was strongly expressed in spindled melanomas and weakly expressed in the epithelioid melanomas. The usual melanoma markers, including HMB45, HMB50, MelanA, and tyrosinase had the reverse pattern, being strongly expressed in virtually all epithelioid melanomas, but rarely expressed in the spindled variants. S100 was unique among the markers in being expressed by both epithelioid and spindled melanomas. Glial fibrillary acidic protein and L1 adhesion protein were expressed moderately, with preferential labeling of the spindled melanomas. The greatest immunophenotypic difference between spindled and epithelioid melanomas was the high abundance of p75NTR expression in spindled melanomas. The functional significance of the high level of p75 neurotrophin receptor expression may contribute to the high predisposition of perineural extension in the desmoplastic subset of spindled melanomas. PMID- 11481519 TI - Interstitial granulomatous drug reaction with a histological pattern of interstitial granulomatous dermatitis. AB - The interstitial granulomatous drug reaction (IGDR) is a novel drug-associated entity, characterized by violaceous plaques with a predilection for skin fold areas. Light microscopically, it resembles the incipient diffuse interstitial phase of granuloma annulare. Differentiating light microscopic features include the absence of complete collagen necrobiosis, the presence of interface dermatitis, and variable lymphoid atypia. The lack of vasculitis rules out the extravascular necrotizing granuloma (Winkelmann granuloma) associated with systemic disease. The differential diagnosis with interstitial granulomatous dermatitis with arthritis as defined by Ackerman et al. has not been studied until now. Our aim was to determine the histologic criteria allowing us to differentiate IGDR without interface dermatitis and lymphoid atypia from interstitial granulomatous dermatitis. We report three patients with IGDR triggered, in two cases by respectively angiotensin convertin enzyme (ACE) inhibitors and furosemide, and in one case by the association of an ACE inhibitor, furosemide, and fluindione. Histologic examination showed a histological pattern of interstitial granulomatous dermatitis. We found a dense, diffuse histiocytic infiltrate distributed interstitially and in palisaded array within the reticular dermis. Eosinophils and some neutrophils were scattered throughout the infiltrate. In some tiny foci, enveloped by histiocytes, thick collagen bundles associated with basophilic nuclear debris or "flame figures" were seen. Vasculitis, interface dermatitis, or lymphoid atypia were absent. Our study allowed us to expand the histological spectrum of IGDR including a histological pattern similar to interstitial granulomatous dermatitis. The lack of degenerated collagen could be a subtle clue in favor of interstitial granulomatous dermatitis triggered by a drug. PMID- 11481520 TI - Atypical pilar leiomyoma: cutaneous counterpart of uterine symplastic leiomyoma? AB - Four cases of cutaneous leiomyoma with cytologic atypia are reported. All four cases were adult men ranging in age from 33 to 92 years of age. Clinical diagnoses were diverse and included epidermal inclusion cyst, scar, nevus, dermatofibroma, squamous cell carcinoma, and basal cell carcinoma. Histological examination of all four lesions revealed a relatively circumscribed, mild to focally moderately cellular, dermal proliferation of large, irregularly shaped spindle cells in a fascicular arrangement. Some cells contained large irregular nuclei and abundant cytoplasm; others were multinucleate. Moderate to focal marked pleomorphism and rare mitotic figures were present, raising the possibility of leiomyosarcoma; however, the sparse mitotic activity and low cellularity did not warrant this diagnosis. This study suggests that cutaneous leiomyomas may exhibit bizarre or "symplastic" patterns analogous to their uterine counterparts. PMID- 11481521 TI - Cytomorphometric parameters and the metastatic potential of cutaneous and uveal melanoma: a comparison with prognostic factors. AB - The Mean of the Ten Largest Nuclei (MTLN1) is one of the most important prognostic factors in uveal malignant melanoma. This study was performed to investigate the applicability of nuclear and nucleolar measurements as a prognostic factor for cutaneous melanoma. A routine light microscope (Carl Zeiss: Axiophot EL-Einnsatz; #451888) at 640 times magnification with a Micrometer Eye Piece (Carl Zeiss: #444034) was used to evaluate the correlation of MTLN1 and Mean of Ten Largest Nuclei (MTLN) with the occurrence of metastasis in 58 primary cutaneous melanoma. For uveal melanoma, cytologic classification was used for comparison. Prognostic value was determined by univariate and multivariate linear regression analysis. MTLN1 was the only significant factor for uveal melanoma (p = 0.05). For cutaneous melanoma, all factors were significantly associated with disease progression. MTLN1 was the only one to remain significant (p + 0.027) in multivariate linear regression analysis. Nuclear and nucleolar morphometry are significant prognostic factors for uveal and cutaneous melanoma. PMID- 11481522 TI - Alterations of mismatch repair protein expression in benign melanocytic nevi, melanocytic dysplastic nevi, and cutaneous malignant melanomas. AB - Immunoperoxidase-staining methods were used to examine the expression of hMLH1, hMSH2, and hMSH6 mismatch repair (MMR) proteins in 50 melanocytic lesions. Microsatellite instability (MSI), screened previously in these lesions by polymerase chain reaction-based microsatellite assay, showed low-level microsatellite instability (MSI-L) in 11 of 22 melanocytic dysplastic nevi (MDN) and two of nine primary cutaneous malignant melanomas (CMMs) but not in the benign melanocytic nevi (BN). Mismatch repair proteins were widely expressed in the epidermis and adnexal structures. All lesions showed positive immunoreactivity with a gradual decrease in the MMR staining values during the progression from BN to MDN to CMMs. The average percentage of positively (PP) stained cells for hMLH1, hMSH2, and hMSH6 in BN was 85.50 +/- 1.95, 77.90 +/- 4.50, and 87.11 +/- 1.85, respectively. The PP cell values in CMMs were significantly reduced as compared with BN (75.22 +/- 3.57, p= 0.01; 56.11 +/- 8.73, p= 0.02; 65.22 +/- 6.47, p = 0.0002 for hMLH1, hMSH2, and hMSH6, respectively). No comparable significant difference was found between microsatellite stable and MSI-L lesions (p = 0.173, p = 0.458, and p = 0.385), suggesting a lack of correlation between MMR expression and MMR function. There was a direct correlation between PP cell values of hMSH2 and hMSH6 (R = 0.39, p = 0.008), implying that their expression could be regulated by a common mechanism. Thus, an important finding of these studies was the reduction of MMR protein levels in CMMs; whether this reflects underlying genetic or epigenetic mechanisms is still to be determined. PMID- 11481523 TI - Reactive angioendotheliomatosis secondary to dermal amyloid angiopathy. AB - Reactive angioendotheliomatosis (RAE) is a rare benign cutaneous vascular proliferation characterized by intravascular hyperplasia of endothelial cells and tuft-like proliferation of vessels. A 75-year-old man had erythematous and violaceous macules, some stellate and others arranged in a livedoid pattern, evolving toward necrosis with central areas having an "atrophie blanche" appearance spread on the trunk, inguinal folds, and right thigh. He was on hemodialysis and had a benign monoclonal gammopathy. Cutaneous biopsy revealed RAE characterized by the proliferation of epithelioid and spindle-shaped cells in superficial and middermis lining vascular channels, arranged in clusters, and sometimes displaying an intravascular growth pattern. These cells stained for CD31, CD34, and actin. Interestingly, prominent amyloid deposits were found in the wall of some vessels in deep dermis, often causing obstruction of their lumina. The cause of RAE is unknown, but it can be associated with infections, antiphospholipid syndrome, dysglobulinemia, cryoproteinemia, and lower extremities arteritis, and it may occur near arteriovenous fistulas. In this patient, we believe that RAE was caused by obliteration of dermal vessels by amyloid deposits. Indeed, it is thought that RAE could be caused by ischemia secondary to vascular obstruction. This is the first reported patient with RAE associated with amyloid deposits. PMID- 11481524 TI - Median raphe cyst of the penis: a report of two cases with immunohistochemical investigation. AB - Penile median raphe cysts are uncommon benign lesions occurring predominantly in the ventral aspect of the glans penis of young men. We observed two cases: those of a 67-year-old patient and a 22-year-old patient. The epithelial lining of the cysts was composed of pluristratified small cells that focally showed rows of columnar cells above the inner surface as well as a monolayered mucinous columnar epithelium. A columnar cell lining predominated in the younger patient. The cytokeratin (CK) immunostaining pattern of the two cysts (CK7(+++), CK13(+++), CK20(-), CAM 5.2(+)) supports the interpretation of a columnar mucinous epithelium undergoing immature urothelial metaplasia. Carcinoembryonic antigen immunostaining positivity of the columnar cells is probably related to the dysembryogenetic cloacal nature of the cysts. Neuroendocrine differentiation of sparse cells interspersed in the pluristratified epithelium was documented by immunohistochemical staining for chromogranin and synaptophysin. Cilia were not identified in the columnar cells by light microscopy or by antidynein immunohistochemistry. PMID- 11481525 TI - Cutaneous signet ring cell carcinoma: a report of a case and review of the literature. AB - Primary cutaneous signet-ring cell carcinoma (CSRCC) is a very unusual but distinctive clinicopathologic entity. It is defined as a diffuse malignant epithelial neoplasia localized in dermis and subcutis without epidermal involvement, showing variable amounts of signet ring cells, without evidence of a visceral adenocarcinoma. We report a case of CSRCC in a 70-year-old man along with its histologic and ultrastructural characteristics, and review of previous cases. We describe their main diagnostic features and discuss its wide differential diagnosis. PMID- 11481526 TI - Pagetoid dyskeratosis of the lips. AB - Pagetoid dyskeratosis is an incidental finding in a variety of lesions of the skin and squamous mucosa. The lesion is considered a selective keratinocytic response in which a small part of the normal population of keratinocytes is induced to proliferate in response to friction. As far as we know, pagetoid dyskeratosis has not been reported in the lips. In this article, we describe the location of the lesion in the lips and its incidence in a group of 90 unselected patients who underwent biopsy or were surgically treated for diverse labial lesions. Histochemical staining and immunohistochemical studies were performed in selected cases. Pagetoid dyskeratosis was found in 38 cases (42.2%) but only in 6 cases (6.7%) the lesion was conspicuous. There was no significant difference between the upper and the lower lip in terms of incidence of the lesion. Labial pagetoid dyskeratosis was more frequent in younger patients (46.7 +/- 25.0 versus 58.5 +/- 20.5; p < 0.05) and in women (chi(2) = 3.89; p < 0.05). Pagetoid cells were more common in suprabasal location and in the labial mucosa. These cells showed positivity for high-molecular weight cytokeratin and negative reaction for low-molecular weight cytokeratin, epithelial membrane antigen, carcinoembryonic antigen, and human papilloma virus. The immunohistochemical profile is different from the surrounding keratinocytes, indicating premature keratinization. The main differential diagnoses include white sponge nevus, leukoedema, oral koilocytoses, hairy leukoplakia, pagetoid squamous cell carcinoma in situ, and extramammary Paget's disease of the oral mucosa. The morphologic features of dyskeratotic pagetoid cells are distinctive and easily recognized as an incidental finding, thus preventing confusion with other important entities including an intraepidermal tumor. PMID- 11481527 TI - Concurrent chronic lymphocytic leukemia cutis and acute myelogenous leukemia cutis in a patient with untreated CLL. AB - Patients who have chronic lymphocytic leukemia (CLL) are known to have a high frequency of second malignant neoplasms. However, acute myelogenous leukemia (AML) occurring concurrent with or after a diagnosis of CLL is extremely rare. In this article we report a case of AML developing in a 55-year-old male with a 6 year history of untreated CLL. The diagnosis was facilitated by touch preparation of a skin punch biopsy specimen. The patient presented with a two-week history of fever, weakness, anasarca, and a skin rash. Physical examination revealed pink to skin-colored firm papules, which coalesced into indurated plaques on his trunk, upper extremities, and face. The lesions, in combination with generalized edema, produced a leonine facies. Touch prep of the biopsy showed medium to large blasts, large monocytoid cells, and numerous small mature lymphocytes, providing the preliminary diagnosis of a second, previously undiagnosed myelomonocytic malignancy in this patient. The initial diagnosis was subsequently confirmed by histologic, cytochemical, immunohistochemical and flow cytometry studies. This is the first reported case of CLL with concurrent AML in which rapid touch prep of a skin punch biopsy facilitated diagnosis. PMID- 11481528 TI - Animal type melanoma: a report of a case with balloon-cell change and sentinel lymph node metastasis. AB - Animal type melanoma is a rare histopathologic variant of melanoma characterized by sheets and nodules of heavily pigmented epithelioid melanocytes that involve the entire thickness of the dermis. This human neoplasm mimics melanocytic neoplasms seen in gray horses and laboratory animals; thus, is termed animal type melanoma. It is quite rare and, with only a few reported cases, its biological behavior is not well understood. We report an example of animal type melanoma on the back of a 27-year-old man. The lesion showed areas of melanoma in situ, which ruled out the possibility of metastatic melanoma. Features of regression were also seen at dermo-epidermal junction and papillary dermis. In some areas, neoplastic melanocytes exhibited a balloon-cell appearance; in others the neoplasm was composed of sheets and fascicles of heavily pigmented epithelioid melanocytes that permeated the entire dermis and extended into the dermal subcutaneous interface, mimicking a cellular blue nevus. Epithelioid melanocytes in deeper areas showed abundant, heavily pigmented cytoplasm and pleomorphic nuclei with prominent eosinophilic nucleoli and some mitotic figures. The neoplastic cells did not show evidence of maturation in deeper areas of the lesion. In some sections, a nodule of heavily pigmented epithelioid melanocytes was seen far from the main bulk of the lesion, at the dermal-subcutaneous interface, raising the possibility of a satellite lesion. A lymphoscintigraphy showed a sentinel lymph node in the right axilla and a subsequent axillary lymphadenectomy demonstrated that the architecture of the sentinel lymph node was effaced by metastatic melanoma. The patient received adjuvant chemotherapy with inteferon alfa-2b and four months after this treatment the patient is alive and well, without evidence of recurrences or additional metastases. PMID- 11481529 TI - A case of juvenile xanthogranuloma arising on a nevus sebaceus. AB - Various epithelial and mesenchymal neoplasms can arise in nevus sebaceus (NS). Juvenile xanthogranuloma arising on an NS has not been reported previously. Juvenile xanthogranuloma, a disease characterized by reactive histiocytic proliferation, could be included in the list of secondary alterations arising on an NS. PMID- 11481530 TI - Everard Home, John Hunter, and cutaneous horns: a historical review. AB - A cutaneous horn is a protrusion from the skin made up of cornified material. These horns can be derived from a variety of epidermal lesions, both benign and malignant. This historical article reviews a number of early instances of cutaneous horns, some reported in the 16th and 17th centuries. The Danish anatomist Thomas Bartholin was the first to have a correct theory of the ethiology of these horny growths, and the English surgeons John Hunter and Everard Home confirmed his findings in the late 18th century. PMID- 11481531 TI - Cutaneous follicle center cell lymphoma, follicular type. AB - This article discusses the clinicopathologic and molecular features of primary cutaneous follicle center cell lymphoma, follicular type. Synthesis of morphologic, immunohistochemical, and molecular studies have clearly characterized this peculiar morphological variant of the cutaneous B-cell lymphomas. Although local recurrences can be frequently observed, the overall prognosis of these patients is very good and extracutaneous dissemination is very rare. PMID- 11481532 TI - Kamino bodies: what they may mean. PMID- 11481534 TI - Overview of modern foldable intraocular lenses and clinically relevant anatomy and histology of the crystalline lens. PMID- 11481537 TI - Anterior capsule opacification. PMID- 11481538 TI - Silicone oil adherence to silicone intraocular lenses. PMID- 11481539 TI - Interlenticular opacification of piggyback intraocular lenses. PMID- 11481540 TI - Calcification on the surface of the Bausch & Lomb Hydroview intraocular lens. PMID- 11481541 TI - Degeneration of ultraviolet absorber material and calcium deposits within the optic of a hydrophilic intraocular lens. PMID- 11481542 TI - Snowflake opacification of poly(methyl methacrylate) intraocular lens optic biomaterial: a newly described syndrome. PMID- 11481543 TI - Posterior capsule opacification. PMID- 11481544 TI - Phakic anterior chamber intraocular lenses. PMID- 11481545 TI - Phakic posterior chamber intraocular lenses. PMID- 11481546 TI - Pediatric cataract surgery and intraocular lens implantation: current techniques, complications, and management. PMID- 11481547 TI - Evolution of surgical techniques and intraocular lens designs for the developing world. PMID- 11481548 TI - Back muscle contraction patterns of patients with low back pain before and after rehabilitation treatment: an electromyographic evaluation. AB - The aims of this study were to conduct a comparative investigation of muscle function between patients with low back pain (LBP) and healthy persons, and to determine whether intensive rehabilitation can change back muscle contraction synergy. Twenty healthy persons and 20 patients with chronic LBP were asked to perform symmetrical and asymmetric tasks. The patients with LBP were tested in the weeks immediately before and after 12 weeks of LBP rehabilitation. Tasks include "carrying" weights up and down and with a 45 degrees left rotation. Eight channel surface electromyographic electrodes were placed on the surface of paraspinal muscles over the lumbar region. Correlations between the right and left corresponding muscles and between values before and after treatment were determined. Lifting capacity for patients with LBP were also measured before and after treatment. Results from electromyographic profiles showed that the muscle activity strategies varied between healthy persons and patients with LBP. The correlation coefficients for spinal muscles have shown very reproducible intrasubject muscle contraction synergies. Unbalanced electromyographic patterns found in patients with LBP given symmetrical tasks were not affected by rehabilitation treatment. PMID- 11481549 TI - Effects of lifestyle and work-related physical activity on the degree of lumbar lordosis and chronic low back pain in a Middle East population. AB - The relationship between the degree of lumbar lordosis and chronic low back pain (LBP) has long been speculated. It is postulated that prolonged sitting and sedentary lifestyle might change the degree of lumbar lordosis and cause LBP. The purpose of this study was to determine the effects of lifestyle, exercise, work setting, work intensity, and other demographic factors such as age, height, weight, and gender on the degree of lumbar lordosis and occurrence of LBP. Eight hundred forty subjects between ages 20 and 65 years were equally categorized into four groups: normal male, normal female, males with LBP, and females with LBP. A questionnaire was used to obtain information about the subject's lifestyle, work setting, level of exercise, and work-related physical activity. A flexible ruler was used to measure lumbar lordosis in all subjects. The average degree of lumbar lordosis for all subjects was 37 degrees +/- 13 degrees. Females had greater lumbar lordosis (42 degrees +/- 15 degrees ) than males did (32 degrees +/- 10 degrees ). There was no significant difference in the degree of lumbar lordosis in subjects with different lifestyle (p = 0.97), level of physical activity (p = 0.36), work setting (p = 0.5), and with or without LBP (p = 0.28). The degree of lumbar lordosis was positively related with the number of pregnancies (p = 0.04, r = 0.25), age (p = 0.02, r = 0.1) and height (p = 0.0001, r = 0.31) and negatively related with weight (p = 0.04, r = 0.06) of the subjects. The likelihood of developing LBP was significantly higher in the subjects who had high work-related physical activity (p = 0.03) and those who exercised less often (p = 0.008). We found no significant relationship between LBP occurrence and the degree of lumbar lordosis (p = 0.68), work setting (p = 0.15), height (p = 0.08), weight (p =0.06), and age (p = 0.67) of the subjects. The degree of lumbar lordosis was not different between normal subjects and those with LBP. Lumbar lordosis was not affected by lifestyle, level of physical activity, or type of work setting. Although these factors have not been found to affect the degree of lumbar lordosis, some affected the occurrence of LBP. This finding indicates that the effect of these factors on LBP involves mechanisms other than changing the degree of lumbar lordosis. PMID- 11481550 TI - Serial changes of herniated intervertebral discs after posterior lumbar discectomy: the relation between magnetic resonance imaging of the postoperative intervertebral discs and clinical outcome. AB - To evaluate the relation between the morphologic changes of postoperative intervertebral discs and the clinical outcome after posterior lumbar discectomy, the size of the bulging disc was analyzed prospectively on serial follow-up magnetic resonance images in 26 randomly selected patients. The bulging of postoperative intervertebral discs involved three patterns of reduction: early reduction (n = 15), gradual reduction (n = 6), and late reduction (n = 5). There was a significant difference in the serial changes of subjective symptoms and neurologic function among the three patterns. A late reduction of postoperative disc bulging could cause late recovery of subjective symptoms and neurologic disturbance. PMID- 11481551 TI - Lumbar posterolateral fusion alone or with transpedicular instrumentation in L4- L5 degenerative spondylolisthesis. AB - We retrospectively reviewed 57 patients with L4--L5 degenerative spondylolisthesis (L4--L5 DS) who underwent posterior decompression and posterolateral fusion of L4--L5 without (Group A) or with (Group B) transpedicular screw instrumentation at least 2 years earlier. The clinical results and fusion rate were similar between Groups A and B, that is, a 72.4% satisfactory outcome with a fusion rate of 82.8% in Group A versus 82.1% satisfactory outcome with a 92.8% fusion rate in Group B. Screw instrumentation reduced postoperative low back pain and resulted in a lordotic slip angle of L4- L5. However, in patients with radiologically excessive segmental motion showing a translational motion of 3 mm or more, flexion angulation of -5 degrees or less, and a slip angle of -5 degrees or less at the site of spondylolisthesis (L4--L5), the kyphotic slip angle (L4--L5) tended to increase after surgery. In the future, in patients with radiologically excessive segmental motion, this point should be considered, and surgical techniques should be evaluated. Our results suggest that the validity of the general addition of screw instrumentation to L4--L5 fusion for L4--L5 degenerative spondylolisthesis is low. PMID- 11481552 TI - Posterior lumbar osteotomy for flat back in adults. AB - Several studies describe sagittal realignment for flat back and related kyphotic decompensation. Official guidelines for sagittal and the frontal realignment have not been developed. In this retrospective study, the authors examined 10 patients with flat back syndrome and treated a related kyphotic decompensation syndrome by posterior wedge osteotomy. The authors' goal was to determine the efficacy of one level osteotomy on sagittal and frontal realignment. The effectiveness of osteotomy performed mainly in L3 or L4 was measured in terms of radiographic sagittal and frontal alignment. Sagittal unbalance was improved in all patients. The correction is related to the restitution of lumbar lordosis (5 degrees of lordosis allows 1 cm of sagittal correction). However, frontal balance is difficult to restore or even to maintain. In one patient, it was worsened and required repeated operation. One-level lumbar osteotomy is a safe procedure to correct sagittal unbalance. Peroperative lordosis correction allows reliable correction planning. The remaining problem is planning for frontal balance correction. PMID- 11481553 TI - Disc and vertebral wedging in patients with progressive scoliosis. AB - A retrospective longitudinal radiographic study of patients with progressive scoliosis was conducted to determine the relative amount of wedging between vertebrae and discs as a function of progression of the scoliosis curve, cause of the scoliosis, and anatomic curve region. Posteroanterior radiographs of 27 patients with idiopathic scoliosis and of 17 patients with scoliosis associated with cerebral palsy were studied. The amount of wedging of vertebrae and discs at the curve apex was measured by the Cobb method and expressed as a proportion of the curve's Cobb angle. On average, the relative amount of vertebral and disc wedging did not differ significantly between initial and follow-up radiographs made after progression of the scoliosis. In both groups of patients, the mean vertebral wedging was more than the disc wedging in the thoracic region; the converse was found in curves in the lumbar and thoracolumbar regions. The patients with scoliosis associated with cerebral palsy had curves that were longer and more commonly in the thoracolumbar and lumbar regions. The relative wedging did not change significantly with curve progression and did not appear to differ by diagnosis. In the management of scoliosis, including small curves, it should be recognized that both the vertebrae and discs have a wedging deformity. PMID- 11481554 TI - Triangulated pedicle screw construct technique and pull-out strength of conical and cylindrical screws. AB - To compare the mechanical effectiveness of a new conical screw design with a conventional cylindrical screw design, the screw insertion time, torque, and pull out strength of single-pedicle screw and triangulated-pedicle screw constructs of each type of screw were compared in human cadaveric vertebral bodies. The time required to insert the conical screws was less than that required for cylindrical screws. Regression analysis revealed a positive correlation between insertion torque and pull-out strength of single and triangulated constructs of each type of screw. The conical screw had a greater increase than the cylindrical screw in the pull-out strength of triangulated pedicle screw constructs. Application of the new conical screw design was significantly faster, and the new screw had better mechanical fixation to the vertebral body than did the conventional cylindrical screw tested. PMID- 11481555 TI - Computed tomography assessment of sacroiliac screw placement relative to the first sacral neuroforamen. AB - The radiographic interpretation of sacroiliac screws relative to the S1 neuroforamen is difficult for orthopedic surgeons and radiologists. Computed tomography (CT) with axial images alone or combined with multiplanar reconstructions are often used to assess screw position. The reliability, reproducibility, and accuracy of orthopedist and radiologist interpretations of axial CT images with and without multiplanar reconstructions was determined using 24 cadaveric hemipelves with known sacroiliac screw position. Interobserver reliability of determining screw position was fair for orthopedists and slight for radiologists regardless of imaging modality or screw composition. Intraobserver reproducibility was moderate for orthopedists regardless of imaging modality or screw type. Reproducibility among radiologists was moderate using axial images of titanium screws and substantial with addition of multiplanar reconstructions. Overall accuracy was similar for orthopedists and radiologists. CT images with multiplanar reconstructions improve accuracy in determining sacroiliac screw position, but not significantly. Current imaging modalities are limited by large inaccuracies and by interobserver and intraobserver variation. PMID- 11481556 TI - Effects of beraprost sodium on canine cauda equina function and blood flow using a chronic spinal cord compression model. AB - Changes in blood flow after chronic compression were observed in 19 dogs after 10 mmHg compression for 1 week before and 1 hour after the intravenous administration of one of three doses of beraprost sodium (BPS; 30 ng x kg(-1) x min(-1), n = 7; 100 ng x kg(-1) x min(-1), n = 7; and 300 ng x kg(-1) x min(-1), n = 5). The speed of blood flow was calculated using a specially designed microscope equipped with a video camera. Dogs treated with BPS had lesser degrees of reduction in their nerve conduction velocity compared with controls. A vascular mechanism of injury likely explains why BPS-treated dogs had a lesser degree of reduction in their nerve conduction velocities compared with the control population. PMID- 11481557 TI - In vivo macrophage recruitment by murine intervertebral disc cells. AB - SUMMARY: An in vivo murine experiment was conducted to measure the capacities of viable intervertebral disc cells to recruit inflammatory cells. The objective was to determine whether compounds secreted from viable cells induce inflammation or whether inflammation in disc herniation simply requires exposure to structural cell or matrix components. Three tissue preparations were inserted into the right lower peritoneal cavity of male mice: tissue with viable annulus fibrosus and nucleus pulposus cells, tissue with viable annulus fibrosus cells, or devitalized annulus fibrosus and nucleus pulposus tissue. Controls included sham-operated and nonoperated groups. Mice were killed 1, 2, or 7 days after surgery. Macrophage recruitment occurred after exposure to viable disc tissue but not after exposure to devitalized disc components; recruitment increased over time. Viable disc cells play a role in the etiology of inflammation in disc herniation. PMID- 11481558 TI - Blood flow measurement in experimental chronic cauda equina compression in dogs: changes in blood flow at various conditions. AB - The purpose of this study was to present a simple method of measurement of blood flow in the nerve root, assess the reliability of this method, and examine the changes of blood flow in chronic cauda equina compression in an experimental setting. A total of 15 dogs were used to determine the blood flow of cauda equina in normal (n = 5), sham (n = 5), and 10 mm Hg compressed cauda equina for 1 week (n = 5). The speed of blood flow was calculated using a specially designed microscope supplied with a video camera. Blood flow in chronic 10 mm Hg compression decreased compared with the sham and normal groups. The kappa value was between 0.75 and 0.96. We conclude that this method might be useful for measurement of blood flow in the nerve root and for confirming the reduction in blood flow of compressed cauda equina. PMID- 11481559 TI - Posterior annular strains during discography. AB - Discography is commonly used in the workup of back disorders. The clinical utility of the test is controversial, and little is known about mechanical changes that may occur in the disc during this exam. To quantify three dimensional deformations of the posterior annulus during discography, and to examine some of the covariates that influence the deformations, displacements of the lumbar posterior annulus were measured during discographic injection for three different spinal positions. Disc bulge and annular strains were calculated from the displacements. The combined effect of disc pressurization, spine position, and location on the disc (lateral versus midline) explained much of the variation in the measured bulges and strains (r(2) = 0.56). Disc pressurization or spine position alone did not always have a significant effect on strains, and the strains and bulges were often influenced by the interactions between position of the spine, location of the disc, and pressurization. In clinical studies of discography, these results suggest that patient position during the examination should be standardized. PMID- 11481560 TI - Intradiscal electrothermal annuloplasty: the IDET procedure. AB - Intradiscal electrothermal annuloplasty is a minimally invasive procedure for the treatment of lumbar degenerative disc disease. This procedure has been used in the lumbar spine of patients who have failed conservative treatment regimens and who might otherwise be candidates for a spinal fusion procedure. As this is a new, recently developed procedure, there is only relatively short-term follow-up data available, and as such, the procedure has not been widely accepted. PMID- 11481561 TI - Vertebral osteomyelitis secondary to Pseudallescheria boydii. AB - Because Pseudallescheria boydii vertebral osteomyelitis is rare and frequently resistant to available antifungal agents, the proper treatment of this lesion has not been defined. To better determine the best treatment of this lesion, the authors evaluated a case P. boydii vertebral osteomyelitis and reviewed the literature. A 48-year-old man had isolated thoracic vertebral osteomyelitis resulting from P. boydii and associated severe thoracic back pain and proximal lower extremity pain and weakness. Magnetic resonance imaging studies revealed continued collapse of the T6--T7 vertebrae despite previous posterior debridement and appropriate antifungal chemotherapy. On admission to the authors' institution, the patient underwent a right thoracotomy, anterior debridement with transthoracic T6--T7 corpectomies and strut grafting, followed by posterior fusion and stabilization with pedicle screws. After operation, the patient's pain, hyperalgesia, and lower extremity symptoms resolved. He was treated with a 12-month course of itraconazole. Imaging and laboratory studies show no evidence of recurrence. P. boydii vertebral osteomyelitis can have devastating neurologic sequelae if not treated properly. The frequent lack of response of this unusual fungal infection to systemic therapy requires frequent serial follow-up examinations. Patients with evidence of progression on imaging studies or neurologic findings should undergo early and aggressive surgical debridement. PMID- 11481562 TI - Occlusion of the left common iliac artery and consecutive thromboembolism of the left popliteal artery following anterior lumbar interbody fusion. AB - We report on a case of occlusion of the left common iliac artery due to arteriosclerosis and consecutive thrombotic occlusion of the left popliteal artery in a 52-year-old man following anterior retroperitoneal interbody fusion of L4--S1. Initial symptoms included leg pain and numbness of the lateral shank, which were thought to be a result of lumbar nerve root irritation from surgery. Diagnosis was not made until 13 days after surgery, when motor deficits were observed. Angiography showed occlusion of the left common iliac artery and thromboembolism of the left popliteal artery. After thromboendarterectomy of the common iliac artery and thrombectomy of the popliteal artery, motor deficits of the left foot were resolved whereas symptoms of pain and sensory deficits continued. Spine surgeons should be aware of this rare complication in cases of postoperative leg pain or of neurologic deficits in the lower extremity after anterior lumbar interbody fusions. PMID- 11481564 TI - Serial changes on quantitative myocardial perfusion SPECT in patients undergoing revascularization or conservative therapy. AB - BACKGROUND: Little is known about changes of myocardial perfusion in patients undergoing coronary revascularization or medical therapy. The purpose of this observational study was to assess the long-term effects of revascularization or conservative therapy on serial quantitative myocardial perfusion single photon emission computed tomography (SPECT). METHODS AND RESULTS: The study population consisted of 421 patients who underwent serial rest thallium-201/stress technetium-99m sestamibi dual-isotope myocardial perfusion SPECT with at least a 1-year interval between the 2 studies and who had abnormal quantitative scan results on the first stress SPECT. The mean interval between scans was 32.7 +/- 15.9 months. Patients were divided into 3 groups according to stress defect extent: group 1 had small stress defects (4%-10%, n = 145), group 2 had intermediate stress defects (>10%-20%, n = 144), and group 3 had extensive stress defects (>20%, n = 132) at baseline. Forty patients in group 1, 44 in group 2, and 54 in group 3 underwent coronary revascularization between 2 SPECT studies; the others had conservative therapy. In group 3 patients with revascularization, stress defect extent and reversible defect extent were remarkably reduced (14.5% +/- 13.6% and 13.1% +/- 12.5%, respectively; both P <.0001), with greater improvement in those patients reporting increased use of cardiac medications; resting defect extent was slightly reduced (1.9% +/- 6.4%, P <.05). In group 3 patients with conservative therapy, a small reduction in stress defect extent was noted (2.3% +/- 8.3%, P <.05). In group 2, there were modest, similar reductions in reversible defect extent in both the patients with revascularization (2.7% +/- 7.7%, P <.05) and those with conservative therapy (1.8% +/- 7.3%, P <.05), as well as a small but significant reduction in stress defect extent in those with conservative therapy (2.1% +/- 8.2%, P <.05). In group 1 patients, no significant changes in stress, rest, or reversible defect extent were found with either therapy. CONCLUSIONS: The findings of this study show that improvement in quantitative myocardial perfusion abnormalities over time occurs in some patients with either revascularization or conservative therapy and suggest that, in patients with extensive defects, greater improvement may be seen in those who undergo revascularization. PMID- 11481565 TI - Prediction of severe coronary artery disease and long-term outcome in patients undergoing vasodilator SPECT. AB - BACKGROUND: Vasodilator perfusion imaging has not been extensively evaluated for predicting severe coronary artery disease (CAD) or long-term prognosis. METHODS AND RESULTS: The goals of this study were to develop a model to predict left main/3-vessel CAD in patients undergoing vasodilator thallium 201 imaging and coronary angiography (angiographic population) and to test the long-term prognostic value of this model in a separate cohort of patients who were not referred for angiography (prognostic population). In the angiographic population (n = 653) the chi2 value of the clinical model (containing the variables age, sex, and prior myocardial infarction) in the prediction of severe CAD was 32. The addition of 3 vasodilator Tl-201 variables (magnitude of ST-segment depression, summed reversibility score, and increased lung uptake) increased the model chi2 value to 114 (P <.001). Only 9% of predicted low-risk patients versus 57% of predicted high-risk patients had severe CAD. In the prognostic population (n = 521) survival rates free of cardiac death or myocardial infarction at 7 years were 91%, 73%, and 51%, respectively, for patient groups predicted to be at low, intermediate, and high risk of severe CAD (P <.001). CONCLUSIONS: Clinical and vasodilator Tl-201 variables can accurately predict the risk of severe CAD. Stress Tl-201 variables add incremental information to clinical variables. The same model also predicts patient outcome. PMID- 11481566 TI - Long-term effect of simvastatin on the improvement of impaired myocardial flow reserve in patients with familial hypercholesterolemia without gender variance. AB - BACKGROUND: Impaired myocardial flow reserve (MFR) in patients with familial hypercholesterolemia (FH) without evidence of ischemia has been reported. However, it has not been clarified whether diminished MFR in such male or female patients with FH can be reversed by simvastatin. METHODS AND RESULTS: Sixteen patients with FH and 16 age-matched control subjects were studied. All patients were proved to have no evidence of exercise stress-induced myocardial ischemia. Baseline myocardial blood flow (MBF) and MBF during dipyridamole administration (MBF [DP]) were measured with positron emission tomography and nitrogen 13 ammonia; MFR was then calculated before and 9 to 15 months after therapy with simvastatin (5-10 mg/day). Total cholesterol level was significantly higher in patients with FH (277 +/- 49.0) than in control subjects (190 +/- 14.9) but was normalized after lipid-lowering therapy (205 +/- 40.3). Baseline MBF was comparable among FH patients before (77.6 +/- 11.6 mL/min/100 g) and after therapy (74.5 +/- 9.62 mL/min/100 g) and control subjects (78.5 +/- 29.9 mL/min/100 g). However, MBF (DP) in FH patients before therapy (178 +/- 50.9 mL/min/100 g) was significantly lower than that in control subjects (282 +/- 148 mL/min/100 g) and was significantly improved after therapy (228 +/- 91.6 mL/min/100 g, P <.05). In fact, there was no statistically significant difference in the MBF (DP) value in FH patients after therapy compared with that in control subjects (P =.09). MFR significantly improved after therapy in patients with FH (3.33 +/- 1.19 vs 2.27 +/- 0.625, P <.01) and was then statistically comparable to that in control subjects (3.54 +/- 1.11). Improvement of MFR was observed whether MBF (DP) before therapy was greater than or less than 200 mL/min/100 g. MFR was improved in both male and female patients with FH. There was a significant relationship between percent change in plasma total cholesterol concentration and percent change in MFR before and after lipid-lowering therapy (r = -0.57, P <.05). CONCLUSIONS: Diminished MFR in patients with FH without evidence of ischemia can be reversed by moderate- to long-term simvastatin therapy without gender variance. PMID- 11481567 TI - Myocardial perfusion in patients with total occlusion of a single coronary artery with and without collateral circulation. AB - BACKGROUND: Previous studies that investigated the effects of coronary collateral circulation on myocardial perfusion were compromised by inclusion of patients with multivessel coronary artery disease, incomplete occlusion, prior myocardial infarction, or a combination of these. In this study we ascertained the relationship between angiographic collateral circulation and myocardial perfusion only in patients with total occlusion of a single coronary artery, in the absence of myocardial infarction or significant stenosis in the other coronary arteries. METHODS AND RESULTS: Seventy-one consecutive patients underwent stress myocardial single photon emission computed tomography within 90 days of angiography. Collateral circulation was present in 49 patients and absent in 22 patients. All but 2 patients had abnormal perfusion by single photon emission computed tomography imaging, with a mean defect size of 19% +/- 12%, and most (83%) had reversible perfusion defects. Defect count activities improved from stress to rest (or redistribution) (45% +/- 13% to 59% +/- 14%, P <.001). Abnormal myocardial perfusion occurred with similar frequency in patients with collateral circulation and in those without it. Total defect size was 19% +/- 12% in patients with and 18% +/- 11% in those without collateral circulation (P = not significant). The extent of reversibility and defect count activity during stress and rest were similar in patients with collateral circulation and in those without it. CONCLUSIONS: In patients with a single-vessel total coronary occlusion and without myocardial infarction, stress-induced myocardial ischemia is almost always present, irrespective of presence or absence of angiographic collaterals. These data lend support to the premise that collateral circulation is rather insufficient to prevent stress-induced ischemia, although it can preserve myocardial viability. PMID- 11481568 TI - Differences due to collimator blurring in cardiac images with use of circular and elliptic camera orbits. AB - BACKGROUND: In cardiac imaging systems, an elliptic acquisition orbit about the patient can be used to enhance resolution of single photon emission computed tomography (SPECT) images by minimizing the distance between the object imaged and the rotating detector system. In this study artifacts from images acquired with the standard circular acquisition are compared with those acquired with various elliptic acquisitions. METHODS AND RESULTS: With the use of elliptic camera orbits of different eccentricities, simulated projection data were generated from a slice through the left ventricle (LV). The projection data included a simulation of the degradation due to the depth-dependent response of the collimator. As is common in many clinical systems, SPECT images were reconstructed with the standard filtered backprojection algorithm without correction for the collimator response. When the ratio of the major-to-minor axis of the acquisition arc is changed from 1 (circular) to 1.5 (elliptic), reconstructed SPECT images show an additional loss of counts (about 10%) in the apical region of the LV. The severity of the apical defect is also dependent on the starting angle of the acquisition arc. When the starting angle is changed from 0 degrees (detector parallel to the major axis of the LV) to 60 degrees, the ratio between the minimum count in the apical region and the maximum count in the left ventricular myocardial wall decreases by as much as 20%. CONCLUSIONS: SPECT image artifacts from elliptic acquisitions are significantly more severe than those from circular acquisitions. Because of the significant difference in images reconstructed from circular and elliptic acquisitions, standardized normal files acquired from circular acquisitions should not be used for comparisons with patient data acquired from elliptic acquisitions. PMID- 11481569 TI - Left ventricular ejection fraction and volumes from gated blood pool tomography: comparison between two automatic algorithms that work in three-dimensional space. AB - BACKGROUND: Two different algorithms, which are fast and automatic and which operate in 3-dimensional space, were compared in the same group of patients to compute left ventricular ejection fraction (LVEF) and volumes from gated blood pool tomography. One method, developed at Cedars-Sinai Medical Center (CS), was dependent on surface detection, whereas the other method, developed at the Free University of Brussels (UB), used image segmentation. METHODS AND RESULTS: Gated blood pool tomograms were acquired in 92 consecutive patients after injection of 740 MBq of technetium 99m-labeled human serum albumin. After reconstruction and reorientation according to the left ventricular long axis, LVEF and left ventricular volumes were measured with the CS and UB algorithms. Measurements of LVEF were validated against planar radionuclide angiocardiography (PRNA) results. The success rates of the algorithms were 87% for CS and 97% for UB. Agreement between LVEF measured with CS and UB (LVEF(CS) = 0.91. LVEF(UB) - 0.85; r = 0.87) and between LVEF measured with CS and PRNA (LVEF(CS) = 1.04. LVEF(PRNA) - 4.75; r = 0.80) and UB and PRNA (LVEF(UB) = 0.98. LVEF(PRNA) + 4.42; r = 0.82) was good. For left ventricular volumes, linear regression analysis showed good correlation between both methods with regard to end-diastolic volumes (r = 0.81) and end systolic volumes (r = 0.91). On average, end-diastolic volumes were similar and end-systolic volumes were slightly higher with CS than with UB. Consequently, significantly lower LVEFs were observed with CS than with UB. CONCLUSIONS: Good correlation was observed between CS and UB for both left ventricular volumes and ejection fraction. In addition, measurements of LVEF obtained with both algorithms correlated fairly well with those obtained from conventional PRNA over a wide range of values. PMID- 11481570 TI - Serial evaluation of fatty acid metabolism in rats with myocardial infarction by pinhole SPECT. AB - BACKGROUND: Iodine 123-labeled 15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (BMIPP) is mainly trapped in the myocardium as triglyceride, depending on the adenosine triphosphate level. Ten percent to 20% of it is metabolized through alpha-oxidation after beta-oxidation; however, the precise mechanism of the regulatory pathways of BMIPP is yet to be clarified. METHODS AND RESULTS: A brief left coronary artery occlusion (10-30 minutes) was performed in 28 male Wistar Kyoto rats. Dual single photon emission computed tomography images of BMIPP and thallium 201 were obtained 3 days and 24 days after the operation. The activities of 3-hydroxyacyl-coenzyme A dehydrogenase (HAD), citrate synthase (CS), and alpha glycerol-phosphate dehydrogenase (GPD) were then measured in both ischemic and nonischemic regions. BMIPP and Tl-201 chloride severity scores were also evaluated conventionally. CS and HAD levels were significantly lower in the ischemic region than in the nonischemic region in the chronic group (CS, 102.9 +/ 28.1 vs 138.7 +/- 33.7 micromol/g/min, respectively, P =.0051; HAD, 54.7 +/- 20.1 vs 78.6 +/- 18.7 micromol/g/min, respectively, P =.0031). There was no difference in GPD between the ischemic and nonischemic regions. The BMIPP severity score had closer inverse relations with HAD (acute, r = -0.82; chronic, r = -0.80) and CS (acute, r = -0.87; chronic, r = -0.81), but not with GPD, than did Tl-201 chloride severity score. CONCLUSIONS: BMIPP imaging correlates well with the activities of HAD and CS, suggesting that a decrease in BMIPP uptake reflects deterioration of both fatty acid metabolism and citrate cycle and shows information other than regional myocardial perfusion. PMID- 11481571 TI - Comparison of Tl-201 with Tc-99m-labeled myocardial perfusion agents: technical, physiologic, and clinical issues. PMID- 11481572 TI - Cellular and molecular basis of inflammatory myocardial disease. PMID- 11481574 TI - Follow-up testing: the beginning of an answer. PMID- 11481573 TI - Regional myocardial mechanics: integrative computational models of flow-function relations. AB - Many cardiac disorders result in regionally altered myocardial mechanics. Although myocardial strain distributions can be measured experimentally and clinically, regional wall stresses must be computed from computational models. Combining these approaches can provide insight into the structural basis of regional dysfunction under conditions such as acute myocardial infarction and ischemia-reperfusion. Recently, 3-dimensional computational models have helped to elucidate the structural basis of the functional border zone adjacent to acutely ischemic myocardium. They have also shown that heterogeneous dysfunction in ischemic-reperfused stunned myocardium does not necessarily imply heterogeneous myofilament injury. Now that computational models are able to reproduce many complex features of the 3-dimensional patterns of regional myocardial deformation observed experimentally, we suggest possible roles for such integrative models in clinical diagnosis. PMID- 11481575 TI - Diagnostic and prognostic applications for vasodilator stress myocardial perfusion imaging and the importance of radiopharmaceutical selection. PMID- 11481576 TI - Beyond the cholesterol profile: monitoring therapeutic effectiveness of statin therapy. PMID- 11481577 TI - Left main coronary artery disease versus catheter-induced vasospasm: elevated right ventricular tracer uptake in a patient with equivocal coronary angiogram results. PMID- 11481578 TI - [Do we need genotype-corrected standard values for the serum-ACE?]. PMID- 11481579 TI - [Corrected normal values for serum ACE by genotyping the deletion-/insertion polymorphism of the ACE gene]. AB - BACKGROUND: In sarcoidosis, serum ACE is widely recognised as a marker of disease activity. Since 1990 a deletion-/insertion polymorphism of the ACE gene is known and a correlation between the genotypes of this polymorphism and serum ACE levels has been observed. Homzygotes for the deletion allele (DD) have the highest levels and homozygotes for the insertion allele (II) the lowest. Heterozygote (DI) persons show intermediate levels. The extent of this influence varies in populations of different ethnic origin. In a large cohort of healthy individuals from North of Germany, genotype-based normal ranges for serum ACE were determined for the population of Germany for the first time. METHODS: In 262 healthy individuals the genotype of the ACE D/I gene polymorphism was determined from genomic DNA by a PCR method. In addition, in serum samples of all these individuals ACE level was measured with a kinetic test. RESULTS: The genotype DD was found in 29.4 % of the individuals examined, the genotype DI in 49.6 % and the genotype II in 21.0 %, respectively. These results are similar to those found in previous investigations in other populations of Central European origin. The mean serum ACE levels (95 % confidence interval) in individuals with the genotypes DD, DI and II are 59.8 U/l (31.8 - 87.8), 47.7 U/l (18.6 - 76.8) and 32.2 U/l (13.7 - 50.7), respectively. Without taking the genotype into account, the average value is 48.0 U/l (15.0 - 80.9). Differences between all genotype groups are highly significant (p < 0.0001). CONCLUSIONS: In sarcoidosis patients, the determination of this ACE gene polymorphism once in the course of the disease allows a better interpretation of the serum ACE levels measured. PMID- 11481580 TI - [Epidemiology and costs of lung cancer in Germany]. AB - OBJECTIVE: Lung cancer shows the leading incidence of all cancers among men in the developed world and an increasing incidence among women. We performed a cost of illness study that aimed to assess the economic burden of lung cancer in Germany and to identify the main cost drivers. METHODS: Costs were estimated for the year 1996. In a retrospective analysis we calculated direct and indirect costs based on secondary data from governmental institutions as well as from the pharmaceutical industry. We chose the cost perspective of sickness funds to estimate direct costs. The human capital approach was applied for the calculation of indirect costs. RESULTS: Total estimated costs were DM 8.31 billion per year. The indirect costs of DM 7.40 billion accounted for 89 % of total estimated costs. The most important cost driver of the indirect costs, early death, represented on its own DM 4.85 billion, according to 58 % of total estimated costs. Of the direct costs, 93 % were due to hospitalization, amounting to DM 0.85 billion. CONCLUSIONS: This cost of illness study concerning lung cancer illustrates the outstanding importance of the indirect costs, mostly due to early death, for total costs. Based on these findings and on the leading role of smoking in the etiology of lung cancer, we suggest that studies dealing with the net costs of smoking to society should include indirect costs. PMID- 11481581 TI - [Guideline on diagnostics and treatment of sleep-related respiratory disorders in adults]. PMID- 11481582 TI - [Fatal outcome of a multisystemic sarcoidosis in a 54-year-old patient]. AB - BACKGROUND: Sarcoidosis is a systemic granulomatous disease of unknown etiology in which the immune system is overstimulated. The intrathoracic lymph glands and the lungs are the most common site of involvement but all other organs and systems can be affected. Although the prognosis is mainly favourable there is a small group of patients who are resistant to therapy and will eventually succumb to their disease. There is no causative treatment but corticosteroids are the most effective therapeutic agents for sarcoidosis. If there is resistance to corticosteroids an immunsuppressive combination treatment is available. The immunsuppressive drugs (Azathioprin, Methotrexat, Ciclosporin, Pentoxifyllin, Thaliodomid) and the clinical effectiveness of the combination treatment are still under investigation. CASE REPORT: We describe a 54-year-old patient admitted to our hospital because of abnormal liver function tests. The cause was a sarcoidosis of the liver but we found also an involvement of the lung. The chest x-ray did not show any pathological findings, the lung function tests - VC and FEV1 - were normal but the diffusion capacity (DLCO) was reduced to 60 %. The course of the disease was characterized by a progressive deterioration of the lung function and of the general condition (loss of weight, fatigue, decrease in vitality). In the further course colon, spleen, bone marrow and kidneys were involved and hypercalcemia, pancytopenia and hyperspleenism occur. The treatment consisted in the application of corticosteroids. We started with 30 mg prednison daily and reduced it over a period of nine weeks to 7.5 mg. In the periods of deterioration we increased the doses up to 40 mg daily and achieved a temporary improvement of the general condition and the function tests. But eventually the fatal outcome could not be prevented not even after the removal of the massive enlarged spleen. The patient died four years after the diagnosis was established. The cause of death was an advanced hepatic encephalopathy as a result of extensive granulomatous involvement of the liver by a multisystemic sarcoidosis. CONCLUSION: We conclude that a steroid resistant multisystemic sarcoidosis should be treated early in the course with a combination of two immunsuppressive agents. We recommend Azathioprin in addition to corticosteroids. We refer to the corresponding literature. PMID- 11481583 TI - [The tuberculosis situation in Germany in 1999]. PMID- 11481584 TI - [Isolated cavitary pulmonary nodule and positron emission tomography]. PMID- 11481585 TI - Studies of association between the gene for calpain-10 and type 2 diabetes mellitus in the United Kingdom. AB - Variation in CAPN10, the gene encoding the ubiquitously expressed cysteine protease calpain-10, has been associated with type 2 diabetes in Mexican Americans and in two northern-European populations, from Finland and Germany. We have studied CAPN10 in white subjects of British/Irish ancestry, using both family-based and case-control studies. In 743 sib pairs, there was no evidence of linkage at the CAPN10 locus, which thereby excluded it as a diabetes susceptibility gene, with an overall sib recurrence risk, lambda(S), of 1.25. We examined four single-nucleotide polymorphisms (SNP-44, -43, -19, and -63) previously either associated with type 2 diabetes or implicated in transcriptional regulation of calpain-10 expression. We did not find any association between SNP-43, -19, and -63, either individually or as part of the previously described risk haplotypes. We did, however, observe significantly increased (P=.033) transmission of the less common C allele at SNP-44, to affected offspring in parents-offspring trios (odds ratio 1.6). An independent U.K. case-control study and a small discordant-sib study did not show significant association individually. In a combined analysis of all U.K. studies (P=.015) and in combination with a Mexican American study (P=.004), the C allele at SNP-44 is associated with type 2 diabetes. Sequencing of the coding region of CAPN10 in a group of U.K. subjects revealed four coding polymorphisms-L34V, T504A, R555C, and V666I. The T504A polymorphism was in perfect linkage disequilibrium with the diabetes-associated C allele at SNP-44, suggesting that the synthesis of a mutant protein and/or altered transcriptional regulation could contribute to diabetes risk. In conclusion, we were not able to replicate the association of the specific calpain-10 alleles identified by Horikawa et al. but suggest that other alleles at this locus may increase type 2 diabetes risk in the U.K. population. PMID- 11481586 TI - A genomewide screen for autism: strong evidence for linkage to chromosomes 2q, 7q, and 16p. AB - Autism is characterized by impairments in reciprocal communication and social interaction and by repetitive and stereotyped patterns of activities and interests. Evidence for a strong underlying genetic predisposition comes from twin and family studies, although susceptibility genes have not yet been identified. A whole-genome screen for linkage, using 83 sib pairs with autism, has been completed, and 119 markers have been genotyped in 13 candidate regions in a further 69 sib pairs. The addition of new families and markers provides further support for previous reports of linkages on chromosomes 7q and 16p. Two new regions of linkage have also been identified on chromosomes 2q and 17q. The most significant finding was a multipoint maximum LOD score (MLS) of 3.74 at marker D2S2188 on chromosome 2; this MLS increased to 4.80 when only sib pairs fulfilling strict diagnostic criteria were included. The susceptibility region on chromosome 7 was the next most significant, generating a multipoint MLS of 3.20 at marker D7S477. Chromosome 16 generated a multipoint MLS of 2.93 at D16S3102, whereas chromosome 17 generated a multipoint MLS of 2.34 at HTTINT2. With the addition of new families, there was no increased allele sharing at a number of other loci originally showing some evidence of linkage. These results support the continuing collection of multiplex sib-pair families to identify autism susceptibility genes. PMID- 11481587 TI - The significance of not finding a gene. AB - As more investigators conduct extensive whole-genome linkage scans for complex traits, interest is growing in meta-analysis as a way of integrating the weak or conflicting evidence from multiple studies. However, there is a bias in the most commonly used meta-analysis linkage technique (i.e., Fisher's [1925] method of combining of P values) when it is applied to many nonparametric (i.e., model free) linkage results. The bias arises in those methods (e.g., variance components, affected sib pair, extremely discordant sib pairs, etc.) that truncate all "negative evidence against linkage" into the single value of LOD = 0. If incorrectly handled, this bias can artificially inflate or deflate the combined meta-analysis linkage results for any given locus. This is an especially troublesome problem in the context of a genome scan, since LOD = 0 is expected to occur over half the unlinked genome. The bias can be overcome (nearly) completely by simply interpreting LOD = 0 as a P value of 1divided by 2ln(2) is approximately equal to .72 in Fisher's formula. PMID- 11481589 TI - Research questions about the nursing labor supply: participation, wages, and pipeline issues. PMID- 11481588 TI - Paternal population history of East Asia: sources, patterns, and microevolutionary processes. AB - Asia has served as a focal point for human migration during much of the Late Pleistocene and Holocene. Clarification of East Asia's role as a source and/or transit point for human dispersals requires that this region's own settlement history be understood. To this end, we examined variation at 52 polymorphic sites on the nonrecombining portion of the Y chromosome (NRY) in 1,383 unrelated males, representing 25 populations from southern East Asia (SEAS), northern East Asia (NEAS), and central Asia (CAS). The polymorphisms defined 45 global haplogroups, 28 of which were present in these three regions. Although heterozygosity levels were similar in all three regions, the average pairwise difference among haplogroups was noticeably smaller in SEAS. Multidimensional scaling analysis indicated a general separation of SEAS versus NEAS and CAS populations, and analysis of molecular variance produced very different values of Phi(ST) in NEAS and SEAS populations. In spatial autocorrelation analyses, the overall correlogram exhibited a clinal pattern; however, the NEAS populations showed evidence of both isolation by distance and ancient clines, whereas there was no evidence of structure in SEAS populations. Nested cladistic analysis demonstrated that population history events and ongoing demographic processes both contributed to the contrasting patterns of NRY variation in NEAS and SEAS. We conclude that the peopling of East Asia was more complex than earlier models had proposed-that is, a multilayered, multidirectional, and multidisciplinary framework is necessary. For instance, in addition to the previously recognized genetic and dental dispersal signals from SEAS to NEAS populations, CAS has made a significant contribution to the contemporary gene pool of NEAS, and the Sino Tibetan expansion has left traces of a genetic trail from northern to southern China. PMID- 11481590 TI - Anxiety and postoperative pain in children who undergo major orthopedic surgery. AB - Anxiety and pain are major concerns not only for children who undergo surgery, but also for their parents and health care professionals. A convenience sample of 74 adolescents who underwent major orthopedic surgery for repair of idiopathic scoliosis and their parents was used to investigate the relationships among children's and parents' preoperative and postoperative anxiety and children's postoperative pain. Age-appropriate versions of Spielberger's State-Anxiety scales measured children's and parents' anxiety, and a visual analog scale assessed children's pain intensities. Children's state anxiety increased from preoperative to postoperative levels, and their postoperative anxiety levels positively related to their pain intensities on days 2 and 4 following the operation. Parents' anxiety decreased from preoperative to postoperative levels, and their postoperative anxiety positively related to their children's postoperative anxiety. Studying both parents and children helped to explain the variance in children's self-reported anxiety. Parents' emotional states are important indicators of children's emotional states and, subsequently, their pain experience. The results of this study suggest that allowing children to assist in the assessment of their postoperative pain may help health care professionals better understand the subjective component of pain. The findings also emphasize the importance of including parents in future studies in which the aim is to understand children's behavioral responses and recovery outcomes. PMID- 11481591 TI - Quantitative and qualitative predictors of development for low-birth weight infants of Latino background. AB - Despite the large number of Latino infants and children in the United States, little Latino-specific data are available for these children. For this study a wide variety of factors that influence the cognitive and motor development of 123 high risk infants from low socioeconomic status Latino families were examined. Because there is not a consensus on the most predictive components of development in this population, a combination of objective and subjective measures were used. The objective measures used were the Home Observation for the Measurement of the Environment (HOME), the Nursing Child Assessment Teaching Scale (NCATS), the Maternal Confidence Questionnaire (MCQ), Bayley scores, and the Parental Stress Index (PSI). Hierarchical multiple regression analysis was used to examine the effect of the objective and subjective measures on the children's mental and motor development. Multiple regression indicated that the Bayley Mental scores at 24 months were predicted from three objective variables (social support, maternal confidence, and mother-infant interaction) and two subjective variables (report of stress and maternal income). For motor development, maternal income, the home environment, and maternal confidence explained most of the variance. The fact that stress, measured subjectively, was the most significant predictor of the cognitive development of Latino low-birth weight infants points to the fact that this construct should not be overlooked and should not be measured by standardized objective measures. Furthermore, the subjective report of mothers in regard to the support they receive was not correlated to the objective measure of social support obtained. Our results indicate that certain factors may be better assessed by interview and subjective measures rather than well-tested objective measures. PMID- 11481592 TI - Nurses' motivation to wash their hands: a standardized measurement approach. AB - Handwashing is a simple procedure that is critical to prevention and control of infections, yet many health care workers (HCWs) do not practice hand hygiene according to recommended guidelines. The Handwashing Assessment Inventory (HAI) is a self-report instrument that is designed to measure the motivational schema of HCWs for handwashing. PMID- 11481593 TI - Assessment of dietary intake of preschool children living in a homeless shelter. AB - Families with young children are the fastest-growing group among the homeless population. This study was undertaken by nursing students and faculty to learn more about what homeless preschool children were fed and what they ate at one family shelter in the Southwest. Results from the study were shared with the entire shelter staff. Mothers who participated in the study were given information on age-appropriate food preparation and servings. This research reveals the important role nurses can play in documenting and teaching both shelter staff and homeless mothers more about children's dietary needs and the long-term health outcomes of a proper diet. PMID- 11481594 TI - Anxiety and depression: incidence and patterns in patients after coronary artery bypass graft surgery. AB - The purpose of this longitudinal study was to describe the incidence and patterns of anxiety and depression in older patients who undergo coronary bypass graft surgery and to determine the influence of gender and age on psychological recovery. A sample of 31 patients was assessed pre- and postoperatively at 2 to 3 days and at 2, 4, 8, and 12 weeks. Younger subjects experienced higher anxiety, reaching statistical significance at 2 and 4 weeks. Women had significantly higher trait and state anxiety and a higher, nonsignificant incidence of depression at all times. Younger subjects were more depressed at 2 to 3 days and at weeks 2 and 4. Because women and younger patients are at higher risk for psychological distress, they should be targeted for interventions. PMID- 11481595 TI - Translating the short version of the Perinatal Grief Scale: process and challenges. AB - Non-English-speaking populations may be excluded from rigorous clinical research because of the lack of reliable and valid instrumentation to measure psychosocial variables. The purpose of this article is to describe the process and challenges when translating a research instrument. The process will be illustrated in the project of translating into Spanish the Short Version of the Perinatal Grief Scale, extensively studied in English-speaking, primarily Caucasian populations. Translation methods, errors, and tips are included. Tools cannot be used in transcultural research and practice without careful and accurate translation and subsequent psychometric evaluation, which are essential to generate credible and valid findings. PMID- 11481596 TI - What's new with institutional review boards? PMID- 11481597 TI - Interactive health communication technology: where do clinical nursing interventions fit into the picture? PMID- 11481598 TI - Adult respiratory distress syndrome among blunt and penetrating trauma patients: demographics, mortality, and resource utilization over 8 years. AB - PURPOSE: The purpose of this study was to compare demographics, resources used, and mortality rates among a subset of trauma patients (blunt versus penetrating) who developed adult respiratory distress syndrome (ARDS) and over two time periods, 8 years apart. PATIENTS AND METHODS: This retrospective observational study was based on an analysis of the computerized Trauma Registry of the Shock Trauma Center of the University of Maryland Medical Systems. All trauma patients with ARDS admitted to a 48-bed trauma intensive care unit (ICU) at a Level I Trauma Center during two time periods of 3 consecutive years each were considered: from January 1, 1985, to December 31, 1987 (G1), and January 1, 1993, to December 31, 1995 (G2). Data were collected in the two time periods on demographics, severity (Revised Trauma Score), injury-admission delay time, first 24-hour transfusion fluids and blood, septic and organ system failure complications, life-support techniques, ICU mortality, and length of stay (LOS). The independent variables were the age, type of trauma, severity scores, transport time, fluid therapy, infectious and organ system failure complications, and life-support techniques. The dependent variable was ICU mortality. RESULTS: A total of 1,108 patients satisfied the entry criteria, 486 in period G1 and 552 in period G2; 929 (89.5%) suffered blunt trauma and 109 (10.5%) penetrating trauma. Mean age was lower for the penetrating trauma populations. There were no significant differences in ICU mortality between the blunt and penetrating trauma patients with ARDS. A significant decrease in ICU mortality was observed between the time periods studied among both blunt (29.5% vs. 21.7%, P <.001) and penetrating trauma patients (25.7% vs. 18.9%, P <.001). A similar rate of renal, hematologic, and cardiovascular organ system failure was observed for both blunt and penetrating trauma patients. Resource utilization measured by ICU LOS and time on mechanical ventilation was also similar in the two groups. The multivariate analysis showed that ICU mortality was related to age, RTS-measured severity, and time period (G1 mortality 1.68-fold that in G2). Renal and hematologic failures entered a second analysis, replacing the time period variable (G2). CONCLUSIONS: ARDS in patients with blunt and penetrating trauma showed similar trends in ICU mortality, complications, and resource utilization. The ARDS mortality rate decreased over the time period studied in both blunt and penetrating trauma. PMID- 11481599 TI - Acute respiratory distress syndrome in children with malignancy--can we predict outcome? AB - PURPOSE: The purpose of this study was to delineate early respiratory predictors of mortality in children with hemato-oncology malignancy who developed acute respiratory distress syndrome (ARDS). MATERIALS AND METHODS: We conducted a retrospective chart review of children with malignant and ARDS who needed mechanical ventilation and were admitted to a pediatric intensive care unit from January 1987 to January 1997. RESULTS: Seventeen children with ARDS and malignancy aged 10.5 +/- 5.1 years were identified. Six of the 17 children (35.3%) survived. Sepsis syndrome was present in 70.6% of all the children. Peak inspiratory pressure, positive end-expiratory pressure (PEEP), and ventilation index values could distinguish outcome by day 3. A significant relationship between respiratory data and outcome related to efficiency of oxygenation, as determined by PaO(2)/FIO(2) and P(A-a)O(2), was present from day 8 after onset of mechanical ventilation. CONCLUSIONS: Peak inspiratory pressure, PEEP, and ventilation index values could distinguish survivors from nonsurvivors by day 3. This may assist in early application of supportive nonconventional therapies in children with malignancy and ARDS. PMID- 11481601 TI - Endogenous heparin decreases the thrombotic response to hemorrhagic shock in rabbits. AB - PURPOSE: The purpose of this study was to determine if endogenous heparin release would modulate the hemostatic response to hemorrhagic shock in rabbits. MATERIALS AND METHODS: Anesthetized rabbits (n = 13) underwent hemorrhagic shock (MAP 30-40 mm Hg) for 60 minutes. Blood samples obtained before and 60 minutes after hemorrhagic shock had thrombelasto-graphic variables (R, reaction time [min]; angle, alpha [ degrees ]; and G [dynes/cm(2)]) determined. Hemostatic function was assessed by modified thrombelastography under four conditions: (1) unmodified sample; (2) platelet inhibition with cytochalasin D; (3) heparinase I exposure; and (4) platelet inhibition and heparinase I exposure. RESULTS: Thrombelastographic variable values in samples without platelet inhibition or heparinase exposure did not significantly change after hemorrhage (before hemorrhage: R = 22.01 +/- 0.7 min, alpha 43.6 +/- 1.3 degrees, G 7,089 +/- 379 dyne/cm(2); after hemorrhage: R 22.1 +/- 2.4, alpha 41.6 +/- 3.9, G = 5,662 +/- 564; mean +/- SEM). However, blood samples exposed to heparinase after hemorrhage demonstrated enhanced hemostatic function with thrombelastographic values (R = 13.4 +/- 1.5, alpha 56.0 +/- 3.4, G = 7012 +/- 565) significantly different (P <.05) from samples not exposed to heparinase. Samples with platelet inhibition demonstrated a similar pattern. CONCLUSION: Hemorrhagic shock significantly increased circulating endogenous heparin activity, attenuating the thrombotic response to hemorrhage in rabbits. Heparin-mediated regulation of hemostasis may serve as a protective mechanism in shock states. PMID- 11481600 TI - Soluble Fas and soluble Fas ligand levels in patients with acute hepatic failure. AB - PURPOSE: The Fas ligand (FasL)/Fas system is an apoptosis induction system that plays an important role in homeostasis and biophylaxis. We measured tumor necrosis factor alpha (TNF-alpha), soluble FasL (sFasL), and soluble Fas (sFas) in patients with acute hepatic failure to determine the relation between such failure and apoptosis. MATERIALS AND METHODS: We assayed 21 blood samples from patients with acute hepatic failure and 8 from patients with sepsis but without acute hepatic failure. Serum TNF-alpha, sFas, and sFasL levels were determined by enzyme-linked immunosorbent assay. RESULTS: sFasL levels were significantly higher in the patients with acute hepatic failure than in the patients with sepsis (0.68 +/- 0.42 ng/mL vs. 0 ng/mL, P =.0001). No significant differences were observed in sFas levels between the two groups. A significant correlation was observed between TNF-alpha and sFas levels (r = 0.657, P =.0008); a negative correlation was observed between TNF-alpha and sFasL levels (r = 0.454, P =.038). CONCLUSIONS: Our results suggest that pathologic aggravation of acute hepatic failure are related to changes in the FasL/Fas system and that TNF-alpha and sFasL, in particular, may play hepatoprotective roles. PMID- 11481602 TI - Continuous venovenous hemofiltration improves intensive care unit, but not hospital survival rate, in nonoliguric septic patients. AB - PURPOSE: The purpose of this study was to assess the effect of the early institution of continuous veno-venous hemofiltration on survival rates of nonoliguric, septic patients. MATERIALS AND METHODS: A retrospective study of 48 nonoliguric septic patients with PaO(2)/FIO(2) < or = 250, who were admitted to the General Intensive Care of the Soroka Medical Center. Twenty-six patients were treated with continuous venovenous hemofiltration (CVVH group) and 22 were treated by conventional therapy. The end point of treatment was weaning from mechanical ventilation, adequate oxygenation, and the need for minimal cardiocirculatory support. RESULTS: The study groups were similar in terms of age, gender, percentage of surgical or nonsurgical patients, APACHE II scores, and the Therapeutic Intervention Scoring System (TISS). Baseline serum urea and creatinine levels were similar in the groups, but the PAO(2)/FIO(2) ratio was significantly lower in the CVVH group (150.6 +/- 86 vs. 214.2 +/- 8.9). Twenty of the CVVH patients and 10 of the patients receiving conventional therapy were discharged from the intensive care unit (P =.03), but the hospital discharge rate was only slightly higher in the CVVH group (12 of 26) compared with the conventional therapy group (7 of 22) (P =.145). CONCLUSIONS: In this retrospective uncontrolled study, the mortality rate was considerably lower in nonoliguric septic patients who received continuous venovenous hemofiltration early in the course of the disease. The improved survival rate may be due to the ability of CVVH to eliminate mediators involved in the septic process, thus averting the multiple system organ failure consequent to septic insult. PMID- 11481603 TI - Bradykinin-associated reactions in white cell-reduction filter. AB - PURPOSE: The purpose of this study was to examine the effect of temperature on bradykinin generation during blood transfusion using positively charged (positive filter), negatively charged (negative filter), and neutral (neutral filter) filters. MATERIALS AND METHODS: Whole blood collected from six volunteers at 4 degrees C or 37 degrees C was passed through the positive or negative filter. In six surgical patients during surgery, autologous blood transfusion at 37 degrees C was initiated through the positive filter, and the same transfusion was reintroduced through the negative filter. Whole blood from another six volunteers at 4 degrees or 37 degrees C was passed through the neutral filter. RESULTS: The positive filter did not generate bradykinin at any temperature, whereas the negative filter generated bradykinin by approximately 4,000-fold when warm blood was used but did not at cool blood. Blood pressure decreased and heart rate increased during warm blood transfusion using the negative filter but did not change using the positive filter. Plasma bradykinin levels increased in patients with use of the negative filter. The neutral filter generated bradykinin when warm blood was used but at levels lower than for the negative filter. CONCLUSIONS: Use of negative filter results in the temperature-dependent generation of bradykinin, which becomes a potential anaphylatoxin when warm blood is used. PMID- 11481604 TI - Mother-to-infant transmission of hepatitis C virus. PMID- 11481605 TI - A bioinformatical approach suggests the function of the autoimmune hepatitis target antigen soluble liver antigen/liver pancreas. AB - Antibodies to a soluble liver antigen/liver pancreas (SLA/LP) appear to be highly specific for the diagnosis of autoimmune hepatitis. The SLA/LP target antigen was recently identified as a hitherto unknown gene encoding 474 amino acid residues. The function of this antigen remains unclear, because it does not share sequence homology with proteins of known function stored in any of the publicly accessible databases. Therefore we used a new theoretical method called fold recognition and could show that the SLA/LP sequence is compatible with the architecture of the superfamily of pyridoxal phosphate (PLP; vitamin B6)-dependent transferases. Its function is likely to be that of a serine hydroxymethyltransferase and may be an important enzyme in the thus far poorly understood selenocysteine pathway. PMID- 11481606 TI - Suppression of bile acid synthesis, but not of hepatic cholesterol 7alpha hydroxylase expression, by obstructive cholestasis in humans. AB - Regulation of bile acid synthesis, a key determinant of cholesterol homeostasis, is still incompletely understood. To elucidate the feedback control exerted on bile acid biosynthesis in humans with obstructive cholestasis, 16 patients with bile duct obstruction were studied. In vivo 7alpha-hydroxylation, reflecting bile acid synthesis, was assayed in 13 of them by tritium release analysis. Serum 27 hydroxycholesterol was determined by gas chromatography-mass spectrometry. In a subgroup, hepatic cholesterol 7alpha-hydroxylase mRNA was assayed by real-time polymerase chain reaction (PCR), enzyme activity was determined by isotope incorporation, and microsomal cholesterol content was assayed by gas chromatography-mass spectrometry. Age-matched control subjects were studied in parallel. Hydroxylation rates were lower in cholestatic patients (108 +/- 33 mg of cholesterol per day, mean +/- SEM; controls: 297 +/- 40 mg/d; P <.01). The reduction was proportional to the severity of cholestasis, and synthetic rates were normalized in 4 subjects restudied after resolution of biliary obstruction. Consistent findings were obtained by analysis of serum 7alpha-hydroxycholesterol levels. On the other hand, hepatic cholesterol 7alpha-hydroxylase mRNA, microsomal enzyme activity, and cholesterol content tended to be increased in cholestasis. Finally, serum 27-hydroxycholesterol levels were slightly reduced in cholestatic subjects and were not related with the severity of the disease. Suppression of in vivo bile acid synthesis with no corresponding reduction in tissue 7alpha-hydroxylase expression and activity is consistent with nontranscriptional, posttranslational levels of regulation; these may play a role in the feedback control of bile acid synthesis in particular conditions. Alteration of the alternate biosynthetic pathway seems unlikely according to the present data. PMID- 11481607 TI - Detection of antimitochondrial autoantibodies in immunofluorescent AMA-negative patients with primary biliary cirrhosis using recombinant autoantigens. AB - Antimitochondrial antibodies (AMA) are the serologic hallmark of primary biliary cirrhosis (PBC). However, depending on the clinical laboratory, from 5% to 17% of PBC patients are consistently AMA-negative, using native mitochondrial antigens and a variety of conventional assays including immunofluorescence (IMF) and enzyme-linked immunosorbent assay (ELISA). The major immunoreactive mitochondrial autoantigens are the E2 members of the 2-oxo-acid dehydrogenase complex family, including pyruvate dehydrogenase complex-E2 (PDC-E2), branched chain 2-oxo acid dehydrogenase complex-E2 (BCOADC-E2), and oxo-glutarate dehydrogenase complex-E2 (OGDC-E2); cDNAs of these proteins have now been cloned, sequenced, and their B cell epitopes defined. In the present study, we cloned cDNAs encoding these proteins from human, not bovine, sources, and expressed the recombinant proteins in a newly developed ELISA that employs a unique Escherichia coli buffer, and compared the data with previous assays using both AMA-positive and -negative patients. Using this new assay and our criteria for positive as an optical density (OD) greater than 10 SD above the mean of control sera, the AMA-positive rate of 191 PBC sera was 94% (179 of 191) compared with 84% (161 of 191) by IMF. None of the 316 control sera were reactive. Using our recombinant assays, we focused attention on the 30 IMF-AMA-negative patients. Twenty-two of 30 (73%) of these patients were positive using this new ELISA. The group of 30 IMF-AMA negative/ELISA-positive patients did not differ significantly from a comparable population of IMF-AMA-positive patients with respect to age, sex distribution, liver function tests, elevation of serum IgM, or pathologic stage. PMID- 11481608 TI - Indomethacin prevents the development of experimental ammonia-induced brain edema in rats after portacaval anastomosis. AB - Patients with fulminant hepatic failure (FHF) die with brain edema, exhibiting an increased cerebral blood flow (CBF) at the time of cerebral swelling. Mild hypothermia prevents brain edema in experimental models and in humans with FHF, an effect associated with normalization of CBF. To study the effects of alterations of CBF on the development of brain edema, we administered intravenous (IV) indomethacin to rats receiving an ammonia infusion after portacaval anastomosis. This model predictably develops brain edema and a marked increase in CBF at 3 hours of infusion. Brain water was measured with the gravimetry technique; CBF was monitored with both laser Doppler flowmetry and radioactive microspheres, whereas intracranial pressure (ICP) was monitored with a cisterna magna catheter. Coadministration of indomethacin prevented the increase in CBF seen with ammonia alone (110 +/- 19% vs. -2 +/- 9%) as well as the increase in brain water (80.86 +/- 0.12% vs. 80.18 +/- 0.06%) and the increase in ICP. Plasma ammonia and brain glutamine levels were markedly elevated in the ammonia-infused group and unaffected by indomethacin. However, ammonia uptake by the brain was significantly reduced by indomethacin. Levels of 6-keto-PGF(1alpha), a stable metabolite of prostacyclin, were reduced in the cerebrospinal fluid (CSF) of indomethacin-treated animals. As with mild hypothermia, avoiding cerebral vasodilatation with indomethacin will prevent the development of brain edema in this hyperammonemic model. Cerebral vasoconstriction reduces cerebral ammonia uptake and, if selective to the brain, may be of benefit in FHF. PMID- 11481609 TI - Short-term prognosis in critically ill patients with cirrhosis assessed by prognostic scoring systems. AB - The short-term prognosis of acutely ill patients with cirrhosis is influenced by the degree of hepatic insufficiency and by dysfunction of extrahepatic organ systems. The purpose of this study was to assess and compare the prognostic accuracy of the Child-Pugh classification, the Acute Physiology and Chronic Health Evaluation (APACHE) II system and the Sequential Organ Failure Assessment (SOFA) for predicting hospital mortality in patients with cirrhosis when used 24 hours after admission to a medical intensive care unit (ICU). Prospective data were recorded on 143 patients. Cumulative mortality rates were 36% in the ICU, 46% in the hospital, and 56% at 6-month follow-up. By using the area under receiver operating characteristic (AUROC) curves, the SOFA showed an excellent discriminative power (AUROC 0.94), which was clearly superior to the APACHE II (AUROC 0.79) and the Child-Pugh system (AUROC 0.74). Hospital mortality rates below and above a cutoff of 8 SOFA points were 4% and 88%, respectively (P <.0005). The SOFA score also reflected resource use during the ICU treatment as measured by daily workload and length of stay. The SOFA is an easily applied tool with excellent prognostic abilities and can be used to enhance clinical judgment of prognosis as well as providing patients and families with objective information. PMID- 11481610 TI - Peroxisome proliferator-activated receptor-alpha mice show enhanced hepatocyte proliferation in response to the hepatomitogen 1,4-bis [2-(3,5 dichloropyridyloxy)] benzene, a ligand of constitutive androstane receptor. AB - Previously, we have suggested that liver cell proliferation induced by certain mitogens is dependent on their binding and activation of nuclear receptors of the steroid/thyroid superfamily. More recently, it was shown that absence of the nuclear receptors peroxisome proliferator-activated receptor-alpha (PPARalpha) and constitutive androstane receptor (CAR) completely abolishes the proliferative response of hepatocytes to the mitogenic stimulus exerted by their specific ligands, peroxisome proliferators (PPs) and 1,4-bis[2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP), respectively. Here we show that deletion of the PPARalpha gene accelerates and enhances the proliferative response evoked by the xenobiotic 1,4 bis[2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP), a powerful mouse-liver mitogen and a ligand of the nuclear receptor CAR. Indeed, the number of hepatocytes entering S phase 24 hours after mitogen treatment was much greater in PPARalpha( /-) mice compared with that of wild type mice (labeling indices 21.4% and 7.5%, respectively). Labeling index of hepatocytes from PPARalpha(-/-) mice was found to be higher than that of wild type mice up to 36 hours after treatment, indicating that lack of PPARalpha not only accelerated but also enhanced the overall proliferative response of the liver. The accelerated entry into S phase observed in hepatocytes from PPARalpha(-/-) mice was associated with a very rapid induction of cyclin D1. No major differences between TCPOBOP-treated PPARalpha(-/ ) and wild type mice were observed in the expression of the 2 inhibitors of cyclin/CDKs complexes, p27 and p21. The results suggest that PPARalpha may play a role in modulating CAR-signaling pathways in the cell, in particular those leading to hepatocyte proliferation. PMID- 11481611 TI - Proliferative activity of intrahepatic colorectal metastases after preoperative hemihepatic portal vein embolization. AB - Although hemihepatic portal vein embolization (PVE) has been used preoperatively to extend indications for hepatectomy in patients with colorectal metastases, the effects of this procedure on tumor growth and outcome remain controversial. To address this issue, we assessed the proliferative activity of intrahepatic metastases after PVE and the long-term outcome of this procedure. Eighteen patients with colorectal metastases underwent preoperative PVE between 1996 and 2000 (PVE group). Twenty-nine patients who underwent major hepatic resection without PVE served as control (non-PVE group). The hepatic parenchymal fraction of the left lobe had significantly increased from 38.1 +/- 3.2% to 45.9 +/- 2.9% 3 weeks after PVE (+20.5%, P <.0001). Tumor volume and percent tumor volume had also significantly increased from 223 +/- 89 mL to 270 +/- 97 mL (+20.8%, P =.016) and from 13.7 + 4.3% to 16.2 + 4.9% (+18.5%, P =.014), respectively. There was no apparent correlation between the increase in parenchymal volume and that in tumor volume. The Ki-67 labeling index of metastatic lesions was 46.6 +/- 7.2% in the PVE group and 35.4 +/- 12.6% in the non-PVE group (P =.013). Long-term survival was similar in the PVE and non-PVE groups, however, disease-free survival was significantly poorer in the PVE group than in the non-PVE group (P =.004). We conclude that PVE increases tumor growth and probably is associated with enhanced recurrence of disease. Although PVE is effective in extending indications for surgery, patient selection for PVE should be cautious. PMID- 11481612 TI - Studies of murine schistosomiasis reveal interleukin-13 blockade as a treatment for established and progressive liver fibrosis. AB - In several allergic, autoimmune, and infectious diseases, fibrosis is a major cause of morbidity and mortality. Here, using a model of infection-induced liver fibrosis, we show that interleukin (IL)-13 is required at all stages of Schistosomiasis mansoni infection to induce fibrosis. IL-4 production was preserved in IL-13-deficient mice, yet failed to significantly contribute to the fibrotic response in either acute or chronic infection. Significant fibrosis develops in all infected mice, although the magnitude of the response varies widely in inbred mice. C3H/HeN, BALB/c, and C57BL/6 mice develop high, intermediate, and low levels of fibrosis, respectively. Despite these differences, IL-13 antagonism resulted in a marked amelioration of fibrosis in all strains. The fibrotic mechanism in the high- and low-responder strains was unrelated to their tissue eosinophil or mast cell responses, but did correlate with their patterns of IL-13, IL-10, and interferon gamma (IFN-gamma) mRNA expression. Indeed, severe fibrosis correlated with a high IL-13 and low IFN gamma/IL-10 mRNA response. Because fibrotic diseases are typically progressive disorders, an important issue was to determine whether IL-13 inactivation might be used to treat an established and ongoing fibrotic disease. Here, IL-13 antagonism was highly efficacious, even after fibrosis and the Th2 cytokine response were firmly established. These studies demonstrate the central role played by IL-13 in fibrogenesis and suggest that therapeutic approaches aimed at disrupting the IL-13 pathway will be highly effective at preventing fibrotic disease caused by chronic Th2-mediated inflammatory reactions. PMID- 11481613 TI - Factors affecting liver fibrosis in human immunodeficiency virus-and hepatitis C virus-coinfected patients: impact of protease inhibitor therapy. AB - Hepatitis C virus (HCV)-related liver fibrosis progression is accelerated in human immunodeficiency virus (HIV)-infected patients. The effect of protease inhibitor (PI) therapy on liver fibrosis is unknown. The aim of this work was to analyze the impact of PI therapy on HCV-related liver fibrosis in HIV/HCV coinfected patients. We evaluated in a long-term follow-up retrospective cohort study the influence of antiretroviral therapy containing PI on liver fibrosis in 182 consecutive HIV/HCV coinfected patients. At liver biopsy, 63 patients had received PI and 119 patients had never been treated with PI. Relationships between liver histologic features, age, alcohol consumption, CD4 cell count, HIV RNA load, and antiretroviral regimens were analyzed. Liver fibrosis stage was lower in patients receiving PIs by comparison with patients who had never received PIs (P =.03). The 5-, 15-, and 25-year cirrhosis rates were 2% versus 5%, 5% versus 18%, and 9% versus 27%, respectively, in patients who had received PIs compared with PI-untreated patients (P =.0006). Multivariate analysis identified 4 independent predictors of progression to cirrhosis: absence of protease inhibitor therapy (relative risk [RR] = 4.74, 95% confidence interval [CI], 1.34-16.67), heavy alcohol consumption (> or = 50 g daily) (RR = 4.71, 95% CI, 1.92-11.57), low CD4 cell count (<200/microL) (RR = 2.74, 95% CI, 1.17-6.41), and age at HCV contamination (> or = 20 years) (RR = 2.37, 95% CI, 1.04-5.38). In conclusion, protease inhibitor therapy might not accelerate progression to HCV related cirrhosis. Furthermore, chronic use of antiretroviral therapy containing PI together with reduction of alcohol consumption and maintenance of high CD4 count could have a beneficial impact on liver fibrosis progression in HIV/HCV coinfected patients. PMID- 11481614 TI - Leptin augments inflammatory and profibrogenic responses in the murine liver induced by hepatotoxic chemicals. AB - Lines of evidence suggested a possible link between leptin and hepatic fibrosis; however, whether leptin modulates the fibrogenesis in the liver remains unclear. The purpose of this study, therefore, was to evaluate the effect of leptin on inflammatory and profibrogenic responses in the liver caused by hepatotoxic chemicals. Male C57Bl/6 mice were given carbon tetrachloride (CCl(4)) (0.1 microL/g body weight [BW], intraperitoneally [IP]) and/or recombinant murine leptin (1 microg/g BW, IP) simultaneously, and sacrificed up to 72 hours later. Further, some mice were given thioacetamide (TAA; 200 microg/g BW, IP) and leptin 3 times per week for 4 weeks to evaluate the effect of leptin on chronic fibrogenic responses. A simultaneous injection of leptin enhanced acute CCl(4) induced necroinflammatory and subsequent fibrotic changes in the hepatic lobules. The steady-state messenger RNA (mRNA) levels of alpha1(I) procollagen and heat shock protein 47 (HSP47) in the liver were potentiated when leptin was injected together with CCl(4). Expression of alpha smooth muscle actin (alpha-SMA) in the liver after CCl(4) treatment was also augmented markedly in combination with leptin. Further, leptin increased transforming growth factor beta1 (TGF-beta1) mRNA in the liver 24 hours after acute CCl(4) about 4-fold higher than CCl(4) alone. Moreover, leptin enhanced hepatic fibrosis and induction of alpha1(I) procollagen mRNA caused by chronic TAA administration. Collectively, these findings indicated that leptin augments both inflammatory and profibrogenic responses in the liver caused by hepatotoxic chemicals. It is postulated that the increase in systemic leptin levels enhances up-regulation of TGF-beta1, leading to activation of stellate cells, thereby augmenting the fibrogenic response in the liver. PMID- 11481615 TI - Hydrogen peroxide mediates vascular cell adhesion molecule-1 expression from interleukin-18-activated hepatic sinusoidal endothelium: implications for circulating cancer cell arrest in the murine liver. AB - The mechanism of intrasinusoidal arrest of circulating cancer cells, which is a critical step in liver metastasis, appears to be facilitated by tumor-derived proinflammatory factors that increase sinusoidal cell adhesion receptors for cancer cells. However, how this prometastatic microenvironment is up-regulated remains unknown. Using intrasplenically injected B16 melanoma (B16M) cells, we show that the expression of vascular cell adhesion molecule-1 (VCAM-1) significantly increased in hepatic sinusoidal endothelium (HSE) cells over physiologic baseline within the first 24 hours of metastatic cancer cell infiltration in the liver. This correlated with increased in vitro adhesion of B16M cells to HSE cells isolated from B16M cell-injected mice. In vivo VCAM-1 blockade with specific antibodies before B16M cell injection decreased sinusoidal retention of luciferase-transfected B16M cells by 85%, and metastasis development by 75%, indicating that VCAM-1 expression on tumor-activated HSE cells had a prometastatic contribution. Because VCAM-1 expression is oxidative stress inducible, recombinant catalase was in vivo administered, resulting in a complete abrogation of both VCAM-1 expression and B16M cell adhesion increases in HSE cells isolated from B16M cell-injected mice. Catalase also abrogated the proadhesive response of HSE cells to B16M-conditioned medium (B16M-CM) in vitro, although this did not affect the concomitant release of major proinflammatory cytokines by HSE cells. HSE cells treated with B16M-CM released interleukin (IL) 18 via tumor necrosis factor-alpha (TNF-alpha)-dependent IL-1beta in vitro. In turn, H(2)O(2) production from B16M-CM-treated HSE cells was regulated by IL-18. Thus, liver-infiltrating B16M cells activated their adhesion to HSE through a sequential process involving TNF-alpha-dependent IL-1beta, which induced IL-18 to up-regulate VCAM-1 via H(2)O(2). The pivotal position of H(2)O(2) was further supported by the fact that incubation of HSE cells with nontoxic concentrations of H(2)O(2) directly enhanced VCAM-1-dependent B16M cell adhesion in vitro without proinflammatory cytokine mediation, which emphasizes the key role of oxidative stress in the pathogenesis of liver inflammation and metastasis. PMID- 11481616 TI - CC-chemokine receptor 6 and its ligand macrophage inflammatory protein 3alpha might be involved in the amplification of local necroinflammatory response in the liver. AB - It is not fully understood how antigen-specific or-nonspecific T cells migrate into the liver in various liver diseases. Macrophage inflammatory protein (MIP) 3alpha is a chemokine that is expressed mostly in the liver, and the receptor CCR6 is reported to be expressed on memory T cells. In the present study, we focused on the expression of CCR6 on T cells from peripheral blood (PB) and inflamed liver, and analyzed the involvement of MIP-3alpha and CCR6 in the immunopathogenesis of liver diseases. Intrahepatic T cells showed significantly (P <.0001) higher percentages of CCR6(+)CD4(+) T cells compared with PB, which is significantly (P =.037) correlated with the necroinflammatory response in the liver. Most of intrahepatic CCR6(+)CD4(+) cells were also positive for CCR5, which is known to express on T-helper 1 cells. MIP-3alpha was stained in the cells located near piecemeal necrosis, where dendritic cells (DCs) are often observed, and coculture of activated DCs with apoptotic cells induced MIP-3alpha production from the DCs. These data suggest that MIP-3alpha is produced by periportal DCs and/or macrophages after necroinflammatory response, leading to the recruitment of activated T cells into the liver. This process could be important to augment the local immune response in the livers of various liver diseases. The finding might be important not only for the understanding of immunopathogenesis of liver diseases but also for the therapeutic strategy to control the local immune response in the liver. PMID- 11481617 TI - The mitochondrial permeability transition contributes to acute ethanol-induced apoptosis in rat hepatocytes. AB - Acute ethanol intoxication induces oxidative stress and apoptosis in primary cultured hepatocytes. Oxidative stress can trigger mitochondrial cytochrome c release initiating the mitochondrial pathway of apoptosis. Based on this information, we formulated the hypothesis that ethanol induced oxidative stress causes mitochondrial dysfunction resulting in apoptosis. In the present study, we found that the mitochondrial membrane permeability transition (MPT) is essential for induction of mitochondrial cytochrome c release and caspase activation of ethanol. The short-term incubation with ethanol (50 mmol/L) induced the MPT, cytochrome c release, caspase activation, and apoptosis of cultured rat hepatocytes. Hepatocyte apoptosis was prevented by caspase inhibitors (i.e., Z VAD-fmk, DEVD-cho, and DMQD-cho). An MPT inhibitor, cyclosporin A, also prevented ethanol-induced cytochrome c release, caspase activation, and apoptosis, suggesting that acute ethanol-induced apoptosis is MPT dependent. Ethanol-induced MPT was also attenuated by N'N'-dimethylthiourea (DMTU, a scavenger of hydrogen peroxide, 10 mmol/L) and N-acetyl-cysteine (NAC, an antioxidant, 5 mmol/L). Preventing hepatocyte MPT by DMTU or NAC attenuated cytochrome c release as well as caspase activation, suggesting that ethanol-induced oxidative stress mediates the MPT. Thus, acute ethanol induces MPT via oxidative stress, and the MPT mediates mitochondrial pathway of apoptosis in hepatocytes exposed to acute ethanol. PMID- 11481618 TI - Beneficial effects of silymarin on estrogen-induced cholestasis in the rat: a study in vivo and in isolated hepatocyte couplets. AB - The effect of silymarin (SIL) on 17alpha-ethynylestradiol (EE)-induced cholestasis was evaluated in rats. EE (5 mg/kg, subcutaneously, daily, for 5 days) decreased both the bile-salt-dependent and the bile-salt-independent fractions of the bile flow. The decrease in the former was associated to a reduction in the bile salt pool size (-58%), and this effect was completely prevented by SIL. This compound also counteracted the inhibitory effect induced by EE on HCO(3)(-) but not glutathione output, 2 major determinants of the bile salt-independent bile flow. EE decreased the secretory rate maximum (SRM) of tauroursodeoxycholate, (-71%) and bromosulfophthalein (BSP; -60%), as well as the expression of the BSP canalicular carrier, mrp2; SIL failed to increase mrp2 expression, and had only a marginal beneficial effect on both tauroursodeoxycholate and BSP SRM values. However, the two-compartment model based kinetic constant for BSP canalicular transfer was significantly improved by SIL (+262%). SIL decreased rather than increased CYP3A4, the cytochrome P450 isoenzyme involved in the oxidative metabolism of EE, and had no inhibitory effect on the UDP-glucuronosyltrasferase isoforms involved in the formation of its 17beta-glucuronidated, more cholestatic metabolite. Pretreatment of isolated rat hepatocyte couplets with silibinin, the major, active component of SIL, counteracted the estradiol 17beta-glucuronide-induced decrease in the percentage of couplets secreting apically the fluorescent bile acid analogue, cholyl-lysyl fluorescein. These results show that SIL protects against EE-induced cholestasis by normalizing mainly the decrease in the bile salt pool size and HCO(3)(-) output, and probably by counteracting the cholestatic effect of its cholestatic, glucuronidated metabolite. PMID- 11481619 TI - Protein kinase C-dependent distribution of the multidrug resistance protein 2 from the canalicular to the basolateral membrane in human HepG2 cells. AB - The subcellular localization of hepatobiliary transport proteins directly affects the rate of bile formation, e.g., the conjugate export pump multidrug resistance protein 2 (MRP2) is regulated on a short-term scale by retrieval from and insertion into the canalicular membrane in the liver. This study reports on the effects of protein kinase C on MRP2 localization and activity in human hepatoblastoma HepG2 cells. MRP2 was detected in HepG2 cells by immunocytochemistry and Western blot analysis. Functional activity was assessed by confocal laser scanning microscopy using fluorescent MRP2 substrates. In untreated HepG2 cells MRP2 was almost exclusively localized at the apical membrane. Treatment of HepG2 cells with phorbol-12-myristate-13-acetate (PMA) resulted in a rapid decrease of apically localized MRP2 and a loss of more than 90% of pseudocanaliculi within 4 hours. This was accompanied by a reduced pseudocanalicular secretion of the MRP2 substrate glutathione-methylfluorescein. Interestingly, PMA treatment (1-100 nmol/L) led to the appearance of immunoreactive MRP2 at the basolateral membrane within 30 minutes. This was shown by its colocalization with MRP1, human dipeptidylpeptidase IV (DPPIV), and transfected rat Ntcp. The effects of PMA on MRP2 localization were sensitive to the protein kinase C (PKC) inhibitor Go6850 but insensitive to inhibition of MEK by PD098059. Basolateral MRP2-appearance was not inhibited by cycloheximide or by disruption of microtubules or microfilaments. In rat livers cholestasis was induced by PMA (100 nmol) and MRP2 was detected at the basolateral membrane in some areas, colocalizing with Ntcp. The data suggest that retargeting of canalicular MRP2 to the basolateral membrane due to PKC activation may represent a novel mechanism that may contribute to cholestasis. PMID- 11481620 TI - Up-regulation of basolateral multidrug resistance protein 3 (Mrp3) in cholestatic rat liver. AB - Cholestasis induces down-regulation of multidrug resistance protein 2 (Mrp2, symbol Abcc2), which is localized to the canalicular membrane. Given the overlapping substrate specificities of Mrp2 and multidrug resistance protein 3 (Mrp3, symbol Abcc3), we examined the hypothesis of a different subcellular and lobular localization of these members of the Mrp family in rat liver after bile duct ligation. We raised a polyclonal antibody against rat Mrp3 and detected this protein in the basolateral plasma membrane of hepatocytes surrounding the central veins and of cholangiocytes. The Mrp3 protein level was less than 2% of the expression observed after 72 hours of obstructive cholestasis. After 48 hours of bile duct ligation, the Mrp3 protein was increased and was further enhanced after 72 hours. In 72-hour-cholestatic rat liver Mrp3 was expressed, in addition, in periportal hepatocytes. However, there was a preponderance of Mrp3 in the pericentral area of the liver lobule. In Mrp2-deficient mutant rat liver, the Mrp3 protein expression was most enhanced and its zonation was lost. The Mrp3 immunostaining of cholangiocytes was preserved in cholestatic and in Mrp2 deficient mutant liver. Canalicular Mrp2 decreased and amounted to 34% of normal after bile duct ligation for 72 hours. We conclude that the hepatocellular up regulation of Mrp3 in cholestasis together with cholangiocellular Mrp3 may compensate for the biliary obstruction and impaired canalicular Mrp2 function by clearing cholephilic anionic substances into the blood. PMID- 11481621 TI - Intrahepatic amino acid and glucose metabolism in a D-galactosamine-induced rat liver failure model. AB - A better understanding of the hepatic metabolic pathways affected by fulminant hepatic failure (FHF) would help develop nutritional support and other nonsurgical medical therapies for FHF. We used an isolated perfused liver system in combination with a mass-balance model of hepatic intermediary metabolism to generate a comprehensive map of metabolic alterations in the liver in FHF. To induce FHF, rats were fasted for 36 hours, during which they received 2 D galactosamine injections. The livers were then perfused for 60 minutes via the portal vein with amino acid-supplemented Eagle minimal essential medium containing 3% wt/vol bovine serum albumin and oxygenated with 95% O(2)/5% CO(2). Control rats were fasted for 36 hours with no other treatment before perfusion. FHF rat livers exhibited reduced amino acid uptake, a switch from gluconeogenesis to glycolysis, and a decrease in urea synthesis, but no change in ammonia consumption compared with normal fasted rat livers. Mass-balance analysis showed that hepatic glucose synthesis was inhibited as a result of a reduction in amino acid entry into the tricarboxylic acid cycle by anaplerosis. Furthermore, FHF inhibited intrahepatic aspartate synthesis, which resulted in a 50% reduction in urea cycle flux. Urea synthesis by conversion of exogenous arginine to ornithine was unchanged. Ammonia removal was quantitatively maintained by glutamine synthesis from glutamate and a decrease in the conversion of glutamate to alpha ketoglutarate. Mass-balance analysis of hepatic metabolism will be useful in characterizing changes during FHF, and in elucidating the effects of nutritional supplements and other treatments on hepatic function. PMID- 11481622 TI - Adult hepatitis B vaccination using a novel triple antigen recombinant vaccine. AB - Present hepatitis B vaccines use multidose prolonged regimens, which even healthcare workers at risk do not always complete. Moreover, when vaccination is completed there remain some who fail to achieve adequate protection. The protection of adults at risk could be improved if there were a more potent vaccine and/or a shorter vaccination regimen available. Vaccine-naive adults were randomized to vaccination with either Engerix-B (SmithKline Biologicals, Rixensart, Belgium) or a novel triple antigen (S, pre-S1, and pre-S2) recombinant vaccine (Hepacare; Medeva Pharma Plc, Speke, UK). The primary efficacy parameter was the degree of seroprotection 6 or 7 months (26 +/- 2 weeks) after beginning vaccination. A total of 304 adults entered the study. Of these, 16 failed to complete the study (9 on Hepacare and 7 on Engerix-B). With the Engerix-B standard (0, 1, 6) regimen, 88% of subjects were protected by month 7, whereas with the triple antigen vaccine a 2-dose regimen (0, 1) provided equivalent protection (91%) within 6 months and a 3-dose (0, 1, 6) regimen was significantly superior (98% seroprotected by 7 months after starting vaccination P <.001). With adults at risk for a suboptimal response (i.e., older adults, the obese, men, and smokers) the triple antigen vaccine produced a greater degree of protection. The vaccines had similar safety profiles. Both vaccines were well tolerated. In healthy normal adults, a triple antigen hepatitis B vaccine containing S and pre S antigens produced an enhanced immunologic response and was as effective as a 2- and 3-dose regimen. PMID- 11481623 TI - A potent antiviral effect on hepatitis C viral dynamics in serum and peripheral blood mononuclear cells during combination therapy with high-dose daily interferon alfa plus ribavirin and intravenous twice-daily treatment with interferon beta. AB - Hepatitis C virus (HCV) is known to infect and replicate within peripheral blood mononuclear cells (PBMC), thereby enabling the direct evaluation of antiviral mechanisms by analyzing HCV dynamics in PBMC. To address potential molecular differences associated with distinct antiviral regimens, we studied HCV dynamics in both serum and PBMC in 44 patients with HCV genotype 1b and high viral load who were randomly assigned to the following 4 different treatment groups: 1) combination therapy with 6 MU daily of interferon alfa 2b (IFN-alpha2b) plus 800 mg of ribavirin; 2) monotherapy with 6 MU daily of IFN-alpha2b; 3) monotherapy with twice-daily intravenous administration with 3MU of IFN-beta; and 4) monotherapy with daily intravenous administration with 6 MU of IFN-beta. HCV-RNA levels were measured serially using highly sensitive real-time detection polymerase chain reaction (PCR). HCV dynamics in both the serum and PBMC showed a "biphasic" pattern. The exponential decay slopes of the second phase were significantly higher in the combination or twice-daily dosing regimen groups compared with groups 2 or 4 (0.10 +/- 0.08 vs. 0.02 +/- 0.09 or 0.16 +/- 0.09 vs. 0.02 +/- 0.04 day(-1); P <.05 or P <.0005, respectively). Moreover, the viral half-lives in the second phase were significantly shorter in these groups (73.2 +/- 42.5 vs. 240.1 +/- 120.7 or 56.0 +/- 44.6 vs. 361.6 +/- 293.5 hours; P <.05 or P <.05, respectively). Additionally, the slope of HCV decline in PBMC tended to be higher in the combination regimens, as compared with monotherapy. Taken together, our data on HCV dynamics provide molecular insight into utilization of combination or twice-daily dosing regimens to increase rates of sustained viral eradication of HCV. PMID- 11481624 TI - A dominant hepatitis B virus population defective in virus secretion because of several S-gene mutations from a patient with fulminant hepatitis. AB - There is increasing evidence that certain pathogenic hepatitis B virus (HBV) variants may play a role in the pathogenesis of fulminant hepatitis (FHB). Recently, we isolated from a patient with fulminant recurrent hepatitis B after liver transplantation variants with enhanced replication competence and a possible defect in viral particle secretion. Both viral features may have contributed to the severity of the disease. The aim of this study was to prove the secretion defect of these variants, to analyze the consequences, and to identify the responsible viral mutations. The variant genomes and appropriate wild-type/variant hybrid genomes were functionally characterized after transfection in human hepatoma cells. Two cloned genomes and the polymerase chain reaction (PCR)-amplified mixture of full-length genomes showed a block in viral particle secretion. This was caused by a combination of amino acid changes in the S-protein including the mutation G145R frequently emerging after hyperimmunoglobulin treatment. The mutations induced retention of the surface proteins in an endoplasmic reticulum (ER)-like compartment, but no intracellular accumulation. These data provide evidence for the in vivo existence of a dominant HBV population with a severe defect in viral particle secretion caused by mutations in the S-gene. This viral phenotype in combination with the enhanced replication competence may have contributed to the fulminant clinical course of the infection. PMID- 11481625 TI - A randomized, double-blind trial comparing pegylated interferon alfa-2b to interferon alfa-2b as initial treatment for chronic hepatitis C. AB - This international, randomized, active-controlled, parallel-group, double-blind dose-finding study compared peginterferon alfa-2b (PegIntron) to interferon alfa 2b for the initial treatment of compensated chronic hepatitis C. We randomly assigned 1,219 subjects to receive either the standard three-times-weekly (TIW) interferon alfa-2b dose (3 MIU) or the once-weekly (QW) peginterferon alfa-2b (0.5, 1.0, or 1.5 microg/kg). Subjects were treated for 48 weeks and then followed for an additional 24 weeks. All 3 peginterferon alfa-2b doses significantly (P < or =.042) improved virologic response rates (loss of detectable serum HCV RNA) after treatment and after follow-up, as compared with interferon alfa-2b. Unlike the end-of-treatment virologic response, the sustained virologic response rate was not dose-related above 1.0 microg/kg peginterferon alfa-2b because of a higher relapse rate among patients treated with 1.5 microg/kg peginterferon alfa-2b, particularly among patients infected with genotype 1. All 3 peginterferon alfa-2b doses decreased liver inflammation to a greater extent than did interferon alfa-2b, particularly in subjects with sustained responses. No new adverse events were reported, and the majority of adverse events and changes in laboratory values were mild or moderate. In conclusion, peginterferon alfa-2b maintained (0.5 microg/kg) or surpassed (1.0, 1.5 microg/kg) the clinical efficacy of interferon alfa-2b while preserving its safety profile. The higher rate of virologic response during treatment with 1.5 microg/kg peginterferon alfa-2b in patients infected with genotype 1 and high viral levels warrants further evaluation. PMID- 11481626 TI - Role of hepatitis B, C, and D viruses in dual and triple infection: influence of viral genotypes and hepatitis B precore and basal core promoter mutations on viral replicative interference. AB - The interactions among hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatitis delta virus (HDV) were studied by measuring HBV-DNA and HCV-RNA levels and by determining the influence of viral genotypes and mutations in HBV basal core promoter (BCP) and precore regions. We included 65 consecutive patients, 25 HBV/HCV, 18 HBV/HDV, and 22 HBV/HCV/HDV. Controls consisted of 55 patients with chronic HBV and 55 with chronic HCV infection. HBV-DNA and HCV-RNA levels were lower in coinfections than in single infections (P <.05). HBV/HCV coinfection was associated with lower HBV viremia (8.2 x 10(4) copies/mL) and lower HCV-RNA levels (7 x 10(5) IU/mL), than the corresponding control group (P <.05), with more marked decrease in HBV replication (P <.05). Moreover, in HBV/HCV coinfection and in triple coinfection we observed an inverse relationship between HBV-DNA and HCV-RNA levels (P <.05). HBV/HDV coinfection was associated with lower HBV viremia (2.5 x 10(4) copies/mL) than that found in HBV infection (P <.05). Patients with triple coinfection showed lower HBV-DNA and HCV-RNA levels than control groups (P <.05). Prevalence of precore mutations was lower in HCV coinfections (P <.05). No significant association was observed between HCV-RNA levels and HBV precore mutations, BCP mutations or HBV genotypes, or between HBV DNA levels and HCV genotypes (P <.05). In conclusion, HCV exhibited stronger inhibitory action in the reciprocal inhibition seen in HBV/HCV coinfection. HDV was the dominant virus in HBV/HDV coinfection and in triple coinfection, and had a greater unfavorable influence on HCV than on HBV replication. The reciprocal inhibition of viral replication seemed to be little influenced by the inherent genomic factors studied. PMID- 11481627 TI - Lamivudine treatment is beneficial in patients with severely decompensated cirrhosis and actively replicating hepatitis B infection awaiting liver transplantation: a comparative study using a matched, untreated cohort. AB - Uncontrolled studies have suggested a beneficial effect of lamivudine in patients with decompensated cirrhosis caused by replicating hepatitis B virus (HBV). We analyzed the outcome of lamivudine treatment in 23 consecutive patients with severely decompensated HBV-cirrhosis defined as a Child-Pugh-Turcotte (CPT) score of > or =10, and compared with a historical untreated control group of 23 patients matched for age, gender, and baseline CPT score. Significant clinical response, defined as a decrease in the CPT score by > or =3 points, was observed in 14 of 23 (60.9%) treated patients versus none of the controls (P <.0001). The median change in CPT scores was -3.0 (range, -6 to +3) in the treated group versus +1.0 in the controls (range, -1 to +2) (P =.016). Orthotopic liver transplantation (OLT) was performed in 34.8% of treated patients (median, 3.5; range, 1-32 months), versus 73.9% of controls (median, 3.0; range, 1-14 months) (P =.04). Excluding transplanted patients, there were no deaths in the treated group versus 6 deaths in the control group (P =.009). Time to death or OLT was significantly longer in treated patients than in controls (P <.001). Two patients developed lamivudine resistance after 9 and 12 months, respectively. Our results suggest that lamivudine significantly improves hepatic function in over half of the patients with decompensated cirrhosis and replicating HBV, and may confer a survival advantage. However, the small sample size and the use of a retrospective control cohort preclude drawing definitive conclusions. Expedited OLT remains the only viable treatment for lamivudine nonresponders. PMID- 11481628 TI - Hepatitis C virus-like particles induce virus-specific humoral and cellular immune responses in mice. AB - We have recently described the production of hepatitis C virus-like particles (HCV-LPs) in insect cells that resemble the putative virions. Here we evaluate the humoral and cellular immunogenicity of the virus-like particles with or without viral p7 protein, a small viral polypeptide that resides between the structural and nonstructural regions of the HCV polyprotein and whose function has not been defined. Immunized BALB/c mice developed high titers of anti-E2 antibodies and virus-specific cellular immune responses including cytotoxic T lymphocytes and T helper responses with gamma interferon production. The virus like particles without p7 generated a higher cellular immune response with a more T(H)1 profile than the particles with p7. Immunization of heat-denatured particles resulted in substantially lower humoral and cellular responses, suggesting that the immunogenicity is strongly dependent on particle formation. Administration of CpG oligonucleotide or cationic lipid 3beta-[N-(N',N' dimethylaminoethane)carbamoyl]cholesterol (DC-Chol), two potent adjuvants, did not significantly enhance the immunogenicity of HCV-LPs. Our results indicate that HCV-LPs can induce humoral and cellular immune responses and offer a promising approach to vaccine development. PMID- 11481629 TI - Occurrence of identical hypervariable region 1 sequences of hepatitis C virus in transfusion recipients and their respective blood donors: divergence over time. AB - A total of 240 stored serum specimens from 30 transfusion recipients and 120 blood donors from the Transfusion-Transmitted Viruses Study (TTVS) were evaluated with the objective of establishing transmission of hepatitis C virus (HCV) by specific blood donors. Phylogenetic analysis of hypervariable region 1 (HVR1) and HCV genotyping were performed on the genomic region encoding amino acids 329 to 410. Amino acid distances between HVR1 sequences were calculated by the Kimura formula. Bootstrap analysis of HVR1 sequences provided support for linking recipients to specific donors. Linear regression analysis showed no differences between donor and recipient HVR1 sequences 7.9 weeks posttransfusion, but donor and recipient sequences diverged thereafter (r = 0.690). The initial lag phase in the evolution of HVR1 in the infected recipient was attributed to the time required to mount host immunologic defenses against the virus. Within-recipient divergence in HVR1 was determined from analyses of serial specimens collected within 2 weeks after the alanine transaminase peak, at the end of the original study (1974-1979), and in the follow-up study (1987-present). HVR1 remained invariant over a period of 6.7 to 9.5 days (95% CI) during acute infection. Within-patient divergence in HVR1 increased over a period of 11 to 15 years (r = 0.771), reaching the degree of divergence observed between unlinked subjects. In cases in which transfusion involved more than one HCV subtype, only one of the HCV subtypes established infection in the recipient. Subtype-specific differences in HVR1 were shown. PMID- 11481630 TI - Smoking and hypoxemia caused by hepatopulmonary syndrome before and after liver transplantation. AB - Severe hypoxemia may occur in patients with liver disease as a result of abnormal intrapulmonary vasodilatations (hepatopulmonary syndrome, HPS). Liver transplantation (LT) is the only effective treatment of HPS, with a quite variable delay of improvement of oxygenation. Smoking, by decreasing respiratory nitric oxide (NO), apparently contributed to improved oxygenation in a 44-year old man with alcohol-induced cirrhosis, complicated by HPS, who underwent LT. The patient quit smoking just before LT, when his PaO(2) was 29 mm Hg and exhaled NO (eNO) 28 ppb, a value far above the normal limits (9.6 +/- 3.2 ppb). After LT, oxygenation remained poor and eNO remained high for more than 4 months, when the patient started to smoke again (blood HbCO going up to 5%). At that time eNO decreased to 6 ppb and PaO(2) increased to 67 mm Hg. The strict relationship between eNO and oxygenation observed in this case reinforces the hypothesis that NO is the most important vasodilating mediator in HPS. Smoking may have hastened the resolution of HPS after LT by inhibiting respiratory NO and/or through a generalized impairment of endothelium-dependent vasodilation. PMID- 11481631 TI - Hepatitis B virus mutants and fulminant hepatitis B: fitness plus phenotype. PMID- 11481632 TI - Give me iNOS or give me death. PMID- 11481633 TI - To biopsy or not to biopsy. PMID- 11481634 TI - To biopsy or not to biopsy. PMID- 11481636 TI - Why and how to innovate liver transplantation: a multiple treatment strategy. PMID- 11481637 TI - Patient safety: to do no harm. PMID- 11481638 TI - Perianesthesia care of cardiac surgery patients: a CPAN review. AB - Postoperative nursing management of cardiac surgery patients is considered part of perianesthesia nursing core curriculum by ASPAN. In many hospitals, however, these patients bypass the PACU and are admitted directly to the ICU. For that reason, perianesthesia nurses who do not have cardiothoracic surgical ICU experience need information related to the immediate postoperative routine and nursing care regimen of cardiac surgery patients. Bleeding, cardiac tamponade, low cardiac output syndrome, and dysrhythmias are postoperative complications that are discussed. A brief overview of postoperative nursing management and several "nursing pearls of wisdom" are also presented. PMID- 11481639 TI - Opioid use during the perianesthesia period: nursing implications. AB - Opioids are used extensively for pain management during the perianesthesia period. These compounds exhibit varying degrees of agonism and antagonism at mu, kappa, and Delta opioid receptors. Stimulation of these receptors cause similar and distinctive actions such as analgesia, euphoria, dysphoria, and respiratory depression. It is imperative that perianesthesia nurses have a clear understanding of opioid receptor site physiology so that maximum analgesic effects are obtained while side effects are minimized. PMID- 11481640 TI - Malignant hyperthermia: perianesthesia recognition, treatment, and care. AB - Although relatively uncommon, malignant hyperthermia (MH) can be a life threatening crisis when it occurs intraoperatively or postoperatively. It is imperative that every member of the perianesthesia team knows what to do and works cohesively because it is a true emergency. This article reviews information for perianesthesia nurses, perioperative nurses, and anesthesia providers about agents that trigger MH; early and late symptoms of MH; recommended medications, equipment, and emergency supplies that should be readily available for use; and the recommended protocol for effectively treating MH. OBJECTIVES: -Based on the content of this article, the reader should be able to (1) describe the pathophysiology of malignant hyperthermia (MH); (2) identify the triggering agents of MH; (3) identify patients at risk for MH; (4) describe the early and late signs of MH; (5) state the medication of choice to treat MH; and (6) describe perianesthesia and/or perioperative nursing interventions for treating an MH patient. PMID- 11481641 TI - When does it begin? The surgeon's orientation. PMID- 11481642 TI - The official ASPAN position: ICU overflow patients in the PACU. PMID- 11481643 TI - Another option for pediatric preanesthesia education. PMID- 11481644 TI - Where should we start first--research topics? PMID- 11481646 TI - Kindergarten supply kit. PMID- 11481647 TI - Molecular genetic aspects of solid tumors in childhood. AB - Advances in molecular genetic research in the past 2 decades have led to an increased understanding of the genetic events in the pathogenesis and progression of human malignancies, including those of childhood. A number of pediatric malignancies have served as models for the molecular genetic approach to patients with cancer. These have highlighted the utility of molecular analysis for a variety of purposes including diagnosis, risk stratification and treatment planning, understanding of syndromes associated with cancer and genetic screening, genetic counseling and prophylactic treatment including surgery. It is likely that there soon will be individualized treatment regimens based on the molecular biologic profile of a patient's tumor. In addition, molecular profiling will lead to new drug development designed to induce differentiation of tumor cells, block dysregulated growth pathways, or reactivate silenced apoptotic pathways. This review discusses the molecular genetic aspects of some of the more common pediatric tumors as well as tumors whose pathogenetic mechanisms are particularly instructive. PMID- 11481648 TI - Cancer of the thyroid gland in infants and children. AB - Carcinoma of the thyroid gland is unusual in children and represents only about 3% of pediatric malignancies. Surgical management is the principal method of treatment, but there is considerable controversy regarding exactly how much of the thyroid gland should be removed for adequate treatment. There also is controversy regarding the use of fine-needle aspiration (FNA) in the evaluation of potentially neoplastic thyroid lesions. In this report, the pertinent literature is reviewed regarding these issues. Moreover, this report will discuss recent discoveries that have elucidated some of the molecular biological events responsible for the development of thyroid cancer. PMID- 11481649 TI - Controversies in the management of Wilms' tumor. AB - Wilms' tumor is the most common malignant renal tumor of childhood; it represents 5% to 6% of childhood cancers in the United States. The survival rate of children with Wilms' tumor has improved dramatically, partly due to large multicenter studies conducted by the National Wilms' Tumor Study Group and the International Society of Pediatric Oncology. To ensure optimal patient outcome, the surgical management of these patients must be appropriate. Controversial issues in the management of Wilms' tumor include the value of preoperative chemotherapy; whether pre-resection biopsy is indicated and if so, how this is best performed; indications for partial nephrectomy; the treatment of low-risk patients with surgery only; and the reliability of preoperative imaging to assess the contralateral kidney. PMID- 11481650 TI - Surgical management of neuroblastoma. AB - Neuroblastoma treatment remains challenging but has been advanced by the establishment of clinical and biological variables that determine prognostic risk. Risk-based therapy currently is the hallmark of neuroblastoma treatment. Initially, stage and age were the prime determinants of survival used in clinical practice. The Shimada histopathologic classification added to the former 2 and biochemical markers like the serum ferritin, lactic dehydrogenase, and neuron specific enolase also provided information regarding prognosis. The current era of neuroblastoma therapy has been influenced heavily by advances in molecular biology, most notably the identification of the MYCN oncogene and the application of recombinant DNA methods to identification of chromosomal deletions. Current risk assessment includes age, stage, histopathology, and biochemical markers but also analyses performed on DNA extracted from fresh tumors. This places the onus of obtaining an adequate quantity and quality of fresh neuroblastoma tissue directly on the pediatric surgeon who performs the initial biopsy. PMID- 11481651 TI - Current status of liver tumors in children. AB - Most tumors of the liver in children are malignant. Benign tumors occur most often in infants, are usually vascular, and seldom require surgery. Hepatoblastoma and hepatocellular carcinoma are the most common malignant tumors. Hepatoblastoma affects younger children and hepatocellular carcinoma affects older children and adolescents. Treatment consists of surgical resection before or after chemotherapy, and long-term survival is impossible without resection. Survival rate exceeds 80% in children with hepatoblastoma. Children with hepatocellular carcinoma and other more unusual malignant tumors do not do as well. Current treatment plans are described for both benign and malignant tumors of the liver. PMID- 11481652 TI - Controversies in the management of paratesticular rhabdomyosarcoma: is staging retroperitoneal lymph node dissection necessary for adolescents with resected paratesticular rhabdomyosarcoma? AB - PURPOSE: Use of retroperitoneal lymph node dissection (RPLND) in paratesticular rhabdomyosarcoma (PTRMS) is controversial and has changed over the past 2 decades. The Intergroup Rhabdomyosarcoma Study Group (IRSG) required ipsilateral RPLND (IRPLND) for all patients with PTRMS treated on IRS-III (1984-91), but changed to clinical evaluation of RPLNs using computerized tomography (CT) in IRS IV (1991 through 1997). In IRS-IV, only those patients with identified lymph node involvement on CT required surgical evaluation of the RPLNs. Nodal radiation therapy was administered only to patients with RPLNs recognized as positive; such patients received more intensive chemotherapy as well. Thus, they compared the incidence of recognized RPLN involvement using these 2 different approaches. They then analyzed patient outcome to determine whether this change in management affected outcome. METHODS: Eligible patients with group I or II PTRMS who were treated on IRS III (n = 100) or IRS IV (n = 134) were analyzed. Failure-free survival (FFS) and survival (S) rates were estimated using the Kaplan-Meier method and compared using the log-rank test. RESULTS: There was a significant change in the distribution of patients with group I versus II tumors from IRS-III to IRS-IV (group I, 68% in IRS-III versus 82% in IRS-IV). This was the result of decreased node recognition when CT was used to stage RPLNs in IRS-IV and was most notable for adolescents (>10 years of age). Overall, 3-year FFS was 92% for patients treated on IRS-III and 86% for those treated on IRS-IV (P =.10), whereas survival estimates were 96% and 92%, respectively (P =.30). Adolescents were at higher risk of RPLN relapse than were children (<10 years of age) and their FFS and survival were worse, regardless of IRS protocol. Furthermore, adolescents with recognized group II tumors experienced better 3-year FFS than those with group I tumors on IRS-IV (100% versus 68%, P =.06), most likely as a result of receiving radiotherapy and intensified chemotherapy. CONCLUSIONS: Use of only CT scan evaluation of RPLN in IRS-IV led to a decrease in identification of RPLN involvement in boys who present with localized PTRMS, and a higher rate of regional relapse as compared with IRS-III. Adolescents had much higher likelihood of RPLN disease, and they fared significantly worse than did younger children on both studies. Furthermore, adolescent boys with group I tumors experienced worse FFS than those with Group II tumors on IRS-IV, probably because some patients with group II tumors were not identified by CT imaging and thus received less effective therapy. These data suggest that adolescents should have ipsilateral RPLN dissection as part of their routine staging, and those with positive lymph nodes require intensified chemotherapy as well as nodal irradiation. PMID- 11481653 TI - Ewing's sarcoma/primitive neuroectodermal tumor of the chest wall. AB - Ewing's sarcoma/primitive neuroectodermal tumor is the most common tumor of the chest wall in children and adolescents. It is extremely malignant with a high frequency of both metastatic spread and of local recurrence. Cure requires intensive therapy to control both distant and local disease. Surgery and high dose radiotherapy can achieve equivalent local control; however, radiation is associated with the additional morbidities of second malignancy and a significant adverse impact on both cardiac and pulmonary function. The optimal therapeutic sequence is initial biopsy followed by induction chemotherapy with subsequent resection of the primary tumor. This approach will achieve the lowest incidence of tumor present at the margins of resection and, hence, need for postoperative radiotherapy. The chest wall is a rare site for tumors in children and adolescents. In a series reported from St Jude's Children's Research Hospital, chest wall tumors constituted only 1.8% of the solid childhood tumors. They are primarily mesenchymal in origin and the Ewing's sarcoma/primitive neuroectodermal tumors (PNET) predominate. This report concentrates on the later tumors. They are recognized to be extremely malignant, and cure in those who present with metastatic disease is very difficult to achieve. Recent advances in our understanding of their cytogenetic basis and optimal treatment are presented. PMID- 11481654 TI - Clinical management of anterior mediastinal tumors in children. AB - The majority of anterior mediastinal tumors in children are malignant. The most common tumors in this location are lymphomas, germ cell tumors, and thymic masses. Most require an operation, either a biopsy or a resection, for cure. Because of their size and location, patients with these masses are at significant risk for cardiopulmonary complications when general anesthesia is administered. Of the many clinical, functional, and radiologic criteria used to identify the children at greatest risk for anesthetic complications, the peak expiratory flow rate (PEFR) and the tracheal cross-section area seem to be the most reliable. General anesthesia should not be administered to children if the PEFR and a tracheal cross section area are both less than 50% of predicted values. If both are greater than 50% of predicted values, general anesthesia can be administered safely. An algorithm is proposed for the preoperative evaluation and management of children with anterior mediastinal tumors. PMID- 11481655 TI - Biomarkers as biological indicators of xenobiotic exposure. AB - The presence of a xenobiotic in the environment always represents a risk for living organisms. However, to talk about impregnation there is a need to detect toxicity in the organism, and the concept of intoxication is related to specific organ alterations and clinical symptoms. Moreover, the relationship between the toxic levels within the organism and the toxic response is rather complex and has a difficult forecast because it depends on several factors, namely toxicokinetic and genetic factors. One of the methods to quantify the interaction with xenobiotics and its potential impact on living organisms, including the human being, is monitoring by the use of the so-called biomarkers. They can provide measures of the exposure, toxic effect and individual susceptibility to environmental chemical compounds and may be very useful to assess and control the risk of long-term outcomes associated with exposure to xenobiotic (i.e. heavy metals, halogenated hydrocarbons, pesticides). PMID- 11481656 TI - Allergenicity testing of supermethrin, phenoxyacetic acid and DNCB using in vivo and in vitro modifications of the local lymph node assays, maximization and epicutaneous testing. AB - The purpose of this study was to compare two methods of testing for allergenicity: in vivo and in vitro modifications of local lymph node assays (LLNA) in mice and the maximization and epicutaneous skin tests in guinea pigs as per the Organization for Economic Cooperation and Development (1981). Two pesticides-the synthetic pyrethroid insecticide supermethrin (SM) and the herbicide phenoxyacetic acid (PAA)-were evaluated using this testing battery. 1 Chloro-2,4-dinitrobenzene (DNCB) was selected as a reference allergen for the local lymph node assay. In vitro modification of LLNA proliferative response per standard cell count in lymphocyte cultures derived from treated Balb/c mice did not differ from control mice. Results of the in vivo modification showed that treatment with 50% PAA and 50% SM resulted in a lower proliferation response of lymphocytes in lymph nodes compared with control animals. The vigour of the proliferative response varied more in in vivo modification of LLNA. Stimulation indices were <3, so PAA and SM did not indicate classification as allergens. Lymphocyte proliferation in 1% DNCB-activated lymph nodes was approximately fivefold higher than in those derived from control mice. Proliferation response in vitro calculated as stimulation index was higher in DNCB-treated mice than those observed in vivo, but differences were not dramatic. Auricular lymph node weight and cellularity in mice treated with PAA and SM were similar to controls. The DNCB stimulation index for lymph node cellularity was 5.5. Lymph node weight was three times higher in comparison with controls. In the maximization test in guinea pigs SM and PAA acid resulted in 40% and 50% of animals demonstrating sensitization, respectively. Epicutaneous administration resulted in weaker reaction. Both SM and PAA are mildly strong sensitizers by this battery. PMID- 11481657 TI - High-dose intravenous paraoxon exposure does not cause organophosphate-induced delayed neuropathy (OPIDN) in mini pigs. AB - Organophosphorus compounds are inhibitors of serine hydrolases. Some of these compounds produce, in addition to their high acute toxicity, a more persistent effect: organophosphate-induced delayed neuropathy (OPIDN). The putative molecular entity whose inhibition is thought to be responsible for OPIDN is the neuropathy target esterase (NTE). Although in vitro NTE is resistant to paraoxon (PX), occasional case reports have associated PX with OPIDN. To assess clinically whether or not high-dose i.v. PX causes OPIDN in mini pigs, 14 mini pigs were anaesthesized, intubated and mechanically ventilated. In a first set of experiments eight pigs received 1 mg PX kg(-1) body weight (BW) dissolved in alcohol. Two control animals received alcohol in a corresponding amount. After infusion of PX, survival of the animals during the acute phase of intoxication was achieved by intensive-care support, using appropriate drugs and fluids according to a pre-established protocol. The mini pigs were extubated 1036 +/- 363 min later (mean +/- SD). The pigs were observed prior to PX application and for 6 weeks thereafter for any abnormalities and/or signs of OPIDN, such as leg weakness, ataxia and paralysis. Observations were graded on a scale for three categories (position, motor deficiency, reaction), with a maximal cumulative score of 9. In a second set of experiments (four additional pigs) larger PX doses were used (3, 9, 27 and 81 mg kg(-1) BW). After recovering from general anaesthesia/surgery, within 2 weeks all animals reached the initial score on the scale. It can be concluded that high-dose i.v. PX exposure does not induce OPIDN in mini pigs during the 6-week observation period. PMID- 11481658 TI - Exposure of Rana ridibunda to lead II. Impact of lead on various parameters of liver metabolism of the frog Rana ridibunda. AB - There are no data at present on the impact of lead (Pb) on amphibian metabolism, although declines of amphibian populations due to man-made changes in the environment have been recorded in recent years. We studied the impact of Pb on the liver metabolism of the frog Rana ridibunda by measuring the hepatic levels of glycogen, lactate, total fat, protein and glucose. Liver is one of the primary target organs of Pb bioaccumulation. Frogs were exposed for 4, 10 and 30 days to 14 mg l(-1) of Pb (in the form of Pb(NO(3))(2) dissolved in water) and compared with matched controls. The level of glycogen in the liver increased until day 30. The increase of the metabolite was time-dependent because there was a positive correlation (r = 0.994, P = 0.006) between glycogen concentration and the days of Pb exposure. Lactate concentration declined continuously up to day 30. Liver fat content decreased from day 10 to day 30. Protein concentration declined continuously until day 30. Glucose concentration increased up to day 30. Glycogen concentration was correlated negatively with liver fat content (r = -0.474, P = 0.005), liver protein content (r = -0.562, P = 0.0004) and lactate concentration (r = -0.472, P = 0.005), whereas it was correlated positively with the Pb burden of the liver (r = 0.578, P = 0.0005). The frog appeared to face a metabolic stress over the 30 days of Pb exposure, without being able to control it. We concluded that the increase of liver glycogen concentration was due to gluconeogenesis via lactate and lipolysis. Further experimentation on key gluconeogenic and lipolytic enzymes over the 30 days of Pb exposure would elucidate the mechanisms that may lead to such phenomena. PMID- 11481659 TI - Lead-induced cell death in testes of young rats. AB - Lead is a well-documented testicular toxicant. The present work was planned to study the occurrence of germ cell death after lead administration. Young growing rats were treated with 5, 10 and 20 mg kg(-1) body weight of lead for 2 weeks. Cell death was assessed by employing in situ TUNEL staining, DNA electrophoresis and morphological examination of the tubules. The results showed that Pb induced significant numbers of germ cells to undergo apoptosis in the seminiferous tubules of rats treated with 20 mg kg(-1) body weight. However, DNA fragmentation was not detected at any of the doses. The level of lead accumulation in the testis increased in a dose-dependent manner. PMID- 11481660 TI - In vitro skin absorption and decontamination of sulphur mustard: comparison of human and pig-ear skin. AB - The aim of this study was to evaluate the use of an in vitro skin diffusion cell system as a model for assessing decontaminants against the chemical warfare agent sulphur mustard (SM). The in vitro absorption rates of SM through heat-separated human (157 +/- 66 microg cm(-2) h(-1)) and pig-ear (411 +/- 175 microg cm(-2) h( 1)) epidermal membranes were in agreement with previous in vivo studies that quoted skin absorption rates of 150 and 366 microg cm(-2) h(-1), respectively. Decontaminants (fuller's earth, Ambergard and BDH spillage granules) were ranked in order of effectiveness by measuring the skin absorption rates and the percentage of applied dose of SM that penetrated human and pig-ear epidermal membranes. The effectiveness of fuller's earth measured in this in vitro study using human epidermal membranes was in agreement with a previous in vivo human volunteer study. Similarly, the effectiveness of fuller's earth and Ambergard measured in vitro with pig-ear epidermal membranes was in agreement with a previous in vivo study conducted on rats. However, there was complete disparity in the ranking of decontaminants between human and pig-ear epidermal membranes measured in vitro. Thus, although pig-ear skin may be a relatively good model for predicting the human skin absorption of SM, it is a poor model for testing decontamination systems. The results of this study further validate the use of Franz-type glass diffusion cells containing human epidermal membranes as a model for predicting in vivo human skin absorption. PMID- 11481661 TI - Determination of therapeutic doses of bisquaternary oximes in large animals. AB - This report presents a non-lethal method for estimating a range of therapeutic doses of bisquaternary oximes that serve as antidotes against organophosphorus poisoning. We have estimated therapeutic oxime doses that are equivalent in their relative toxicity rather than selecting arbitrary fractions of their LD(50). Thus, toxic signs of the oximes HI-6, HLo-7, Toxogonin, AB-8 and AB-13 were monitored quantitatively in baboon monkeys and beagle dogs. Using Toxogonin as a reference oxime, a calculated unit of equivalent dose (CED) was defined as the oxime dose equal to the ratio between its minimal toxic dose (MTD) and the therapeutic ratio (TR) of Toxogonin i.e. CED = MTD/TR. Assuming that the tails of dose-response curves of toxicity for bisquaternary oximes are shallow and similar to one another, one could substitute the ED(10) for the MTD. The ED(10) values for bisquaternary oximes were estimated using the log-log model following experimental observations and quantitative scoring of toxic signs in dogs and monkeys. The MTD values then were calculated using the ED(10) values and the experimental therapeutic dose of the reference oxime Toxogonin. The following CED values were obtained for AB-8, AB-13, Toxogonin, HI-6 and HLo-7 in dogs (d) and monkeys (m): 98.7, 74.2, 30.0, 14.5 and 12.1 (d) and 281.9, 232.1, 41.7, 192.9 and 92.9 (m) micromol kg(-1), respectively. The antidotal efficacy of these oximes against poisoning by the nerve agent tabun was determined in dogs and monkeys. These dose-dependent efficacy data were obtained at 0.3 x CED, 1 x CED and 3 x CED of oximes in combination with atropine. These data provide comparative therapeutic values using oxime doses based on their relative toxicity. The highest antidotal efficacy against tabun in dogs was obtained for toxogonin, whereas HLo-7 and AB-13 were most efficacious in monkeys. PMID- 11481662 TI - Protective action of the serine protease inhibitor N-tosyl-L-lysine chloromethyl ketone (TLCK) against acute soman poisoning. AB - Soman-poisoned rats display cholinergic crisis, a systemic mast cell degranulation characteristic of anaphylactic reactions and an excitotoxin-like sequential seizure and neuronal degeneration. The protection of guinea pigs from soman lethality by prophylactic administration of the serine protease inhibitor suramin suggests a possible proteolytic component in soman poisoning. The present study tested the effect of N-tosyl-L-lysine chloromethyl ketone (TLCK), an inhibitor of trypsin-like serine proteases, on soman-induced toxic signs (convulsions, righting reflex) and survival time. Nine control guinea pigs receiving 2 x LD(50) (56 microg kg(-1), s.c.) of soman immediately followed by a therapeutic dose of atropine sulfate (17.4 mg kg(-1) i.m.) experienced severe convulsions, and 8/9 lost the righting reflex. Six of these nine animals expired within 65 min; the three remaining animals survived 24 h to termination of the experiment. When a second group of animals were given TLCK (12 mg kg(-1), i.p.) 30 min prior to a 2 x LD(50) soman challenge and atropine-sulfate therapy, 5/9 experienced convulsions and only 3/9 lost the righting reflex. All nine animals survived beyond 4 h, with six surviving to 24 h. Compared with soman controls, prophylaxis with TLCK significantly prevented the loss of righting reflex (P = 0.05) and enhanced 4-h survival (P = 0.005). Although, convulsions were reduced and 24-h survival was improved in TLCK-treated animals, these results were not statistically significant. The protection from soman toxicity by chemically distinct protease inhibitors such as suramin and TLCK suggests a role for pathological proteolytic pathways in soman intoxication. PMID- 11481663 TI - Oxidative preconditioning affords protection against carbon tetrachloride-induced glycogen depletion and oxidative stress in rats. AB - The rectal insufflation of a judicious dose of ozone, selected from that used in clinical practice, is able to promote oxidative preconditioning or oxidative stress tolerance preventing the hepatocellular damage mediated by free radicals. In order to evaluate the effects of ozone oxidative preconditioning on carbon tetrachloride-mediated hepatotoxicity, the following experimental protocol was designed: group 1 (negative control, sunflower oil i.p.); group 2 (CCl(4) in sunflower oil, 1 ml kg(-1) i.p.); group 3 (15 ozone-oxygen pretreatments at a dose of 1 mg kg(-1) via rectal insufflation + CCl(4) as in group 2); group 4 (ozone control group, 15 ozone-oxygen pretreatments + sunflower oil i.p.). Ozone pretreatment prevented glycogen depletion (as demonstrated by biochemical and histopathological findings) and avoided lactate overproduction associated with the hepatotoxic effects of CCl(4). The administration of CCl(4) increased lipid peroxidation (as measured by thiobarbituric acid-reactive substances) and uric acid levels and inhibited superoxide dismutase activity. All these deleterious effects induced by CCl(4) were prevented by ozone pretreatment. The administration of ozone without CCl(4) (ozone control group) did not produce any changes in the evaluated parameters. Our results showed that ozone treatment, in our experimental conditions, was able to prevent anaerobic glycolysis and oxidative stress induced by CCl(4). PMID- 11481664 TI - Peritoneal toxicity of water: a model of chronic peritonitis caused by osmotic dysequilibrium in rats. AB - The purpose of this work was to determine whether the osmotic dysequilibrium created by intraperitoneal (i.p.) injection of pure water has any permanent, damaging toxic sequelae. Rats were injected i.p. with pure water on five successive days. Necropsies were performed 1 week after the last injection. Necropsies revealed fibrosis of peritoneal surfaces of liver and spleen, similar to the effects of chemical irritants but milder. The severity of the lesions depended on the dose of water and the number of injections. A few minutes of contact with pure water was sufficient to ensure subsequent development of fibrosis. Soon after the initial injury, the inoculum became less hypotonic and then isotonic. Isotonic or moderately hypotonic electrolyte solutions did not produce peritoneal fibrosis but very hypotonic solutions were toxic. Injection of the synthetic compound 48/80, which is known to cause discharge of mast cell granules, did not produce peritonitis, nor was contamination by endotoxin or by blood responsible for the lesions. Injection of water may be a useful method for investigating the role, if any, of mast cells and/or mesothelial cells in the toxic effects of osmotic dysequilibrium. PMID- 11481665 TI - Inhibition and recovery of maternal and fetal cholinesterase enzyme activity following a single cutaneous dose of methyl parathion and diazinon, alone and in combination, in pregnant rats. AB - Pregnant Sprague-Dawley rats (14-18 days of gestation) were treated with a single cutaneous subclinical dose(s) of 10 mg kg(-1) (15% of LD(50)) of methyl parathion (O,O-dimethyl O-4-nitrophenyl phosphorothioate) and 65 mg kg(-1) (15% of LD(50)) of diazinon (O,O)-diethyl O-2-isopropyl-6-methylpyrimidinyl phosphorothioate, and their combination. Animals were sacrificed at 1, 2, 4, 12, 24, 48, 72, and 96 h after dosing. Inhibition of maternal and fetal cholinesterase enzyme activity has been determined. Methyl parathion significantly inhibited maternal and fetal brain acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BuChE) activity within 24 h after dosing. Diazinon and a mixture of methyl parathion and diazinon caused lesser inhibition compared with methyl parathion alone. Recovery of maternal and fetal brain AChE activity was in the order of diazinon > combination of diazinon and methyl parathion > methyl parathion 96 h after dosing. Although fetal plasma BuChE activity recovered to 100% of control within 96 h of application, maternal BuChE activity remained inhibited to 55% and 32% of control 96 h after application of methyl parathion and a mixture of methyl parathion and diazinon, respectively. Following a single dermal dose of methyl parathion, the activity of maternal liver BuChE was 63% of control 2 h after dosing, whereas inhibition of placental AChE or BuChE activity occurred 12 and 1 h following a single dose of methyl parathion, corresponding to activities of 63% and 54% of control, respectively. Diazinon, alone or in combination with methyl parathion, did not inhibit significantly the maternal liver BuChE or placental AChE and BuChE activity. The results suggest that dermal application of a single dose of methyl parathion and diazinon, alone or in combination, has an easy access into maternal and fetal tissues, resulting in inhibition of cholinesterase enzymes. The lower inhibitory effect of the combination of methyl parathion and diazinon might be due to competition of diazinon with methyl parathion for cytochrome P-450 enzymes, resulting in formation of the potent cholinesterase inhibitor methyl paraoxon. The faster recovery of fetal cholinesterase enzymes is attributed to the rapid de novo synthesis of cholinesterase fetal tissues compared with the mother. PMID- 11481666 TI - Lack of adverse effects on fertility of female CD-1 mice exposed to repetitive intravaginal gel-microemulsion formulation of a dual-function anti-HIV agent: aryl phosphate derivative of bromo-methoxy-zidovudine (compound WHI-07). AB - 5-bromo-6-methoxy-5,6-dihydro-3(')-azidothymidine-5(')-(p-bromophenyl) methoxyalaninyl phosphate (WHI-07), a novel bromo-methoxy-substituted aryl phosphate derivative of zidovudine (ZDV), is a potent dual-function contraceptive agent with anti-HIV activity. Its potential for reproductive toxicity was assessed in a series of experiments using CD-1 mice under the conditions of its intended use as an intravaginal microbicide. Female CD-1 mice were exposed intravaginally to a gel-microemulsion formulation containing 0%, 0.5%, 1.0% or 2.0% WHI-07 for up to 13 weeks. On a molar basis, these concentrations represent 1400-5700 times its in vitro spermicidal IC(50) and 1.4-5.7(x10(6)) times its in vitro anti-HIV IC(50). We examined the effects of intravaginally administered WHI 07 on: ovulation efficiency; in vivo fertilization and early embryonic, fetal development; and reproductive outcome, including neonatal survival and pup development. Compound WHI-07 was administered intravaginally during superovulation, organogenesis and prior to mating for 5 and 10 consecutive days and for 13 weeks, respectively. Mice were evaluated for ovulation efficiency and fertilization rate and cleavage 14 and 40 h after human chorionic gonadotropin (hCG) injection, respectively. Pregnant mice were administered 2% WHI-07 intravaginally during gestation days (GD) 6-15 and measures of teratogenicity were evaluated on GD 17. For short-term toxicity study, mice were given intravaginal treatment of gel-microemulsion containing 0%, 0.5%, 1.0% and 2.0% WHI-07 for 13 weeks and then mated with untreated males to evaluate potential reproductive and developmental effects. Repeated intravaginal exposure of mice to 2% WHI-07 had no adverse effects on ovulation response, mean number of eggs recovered or the percentage of eggs fertilized or cleaved. No evidence of reproductive toxicity, fetal toxicity or teratogenicity was found following repetitive intravaginal application of 2% WHI-07 during the period of organogenesis. Furthermore, repeated intravaginal exposure of mice to 0.5-2.0% WHI-07 for 13 weeks had no adverse effect on the subsequent reproductive capability, perinatal outcome or growth and development of the offspring. Compound WHI-07 shows unique clinical potential as a safe, dual-function vaginal contraceptive for curbing mucosal and perinatal HIV transmission. PMID- 11481667 TI - Uncoupling effect of mercuric chloride on mitochondria isolated from an hepatic cell line. AB - A human fetal hepatic cell line (WRL-68) was used as a model to study the damage produced by mercury. The Hg(II) uptake by WRL-68 cells was found to be in a biphasic manner with a rapid initial uptake phase lasting about 5 min, followed by a sustained phase of slower accumulation. Distribution of mercury was studied and mitochondria were found to be the major target for mercury in this cell line (48%), followed by nuclei (38%), cytosol (8%) and microsomes (7%). Mitochondrial morphological damage after mercury treatment was observed by transmission electron microscopy. To determine if the toxic effect of mercury on mitochondrial bioenergetics was direct or indirect, mitochondria were isolated from WRL-68 cells after 1 h of pre-incubation with 0.5 microM HgCl(2). Oxygen consumption was quantified in two sets of experiments: in the presence of classical mitochondrial respiratory inhibitors; and in the presence of oligomycin. No significant difference was found in respiration with classical inhibitors, indicating that mercury does not affect directly the mitochondrial respiratory chain. However, mitochondria of Hg-treated cells were not inhibited when oligomycin was added, probably due to an uncoupling effect. This effect was prevented with dithiothreitol (DTT) treatment. A possible explanation for mercury's effect on mitochondria and its relation with oxidative stress is presented. PMID- 11481668 TI - Bis(4,7-dimethyl and 5-dinitro-1,10-phenanthroline) sulfato-oxovanadium(IV) mediated in vivo male germ cell apoptosis. AB - Oxovanadium(IV) [VO] complexes of 1,10-phenanthroline are a new class of potent apoptosis-inducing cytotoxic agents against human testicular cancer cells in vitro. The present study investigated the in vivo ability of four(bis)-chelated 1,10-phenanthroline [phen] complexes of sulfato-oxovanadium(IV)-VO(phen)(2), VO(Cl-phen)(2), VO(Me(2)-phen)(2) and VO(NO(2)-phen)(2)-with and without substitutions, to induce testicular germ cell apoptosis. Male germ cell loss in mice was measured by determining the epididymal sperm count, testicular weight and histological evaluation of the testes. Repetitive intratesticular injection (7.5 mg kg(-1) testis(-1)) of bis-chelated 1,10-phenanthroline complexes of oxovanadium(IV) with 4,7-dimethyl [VO(Me(2)-phen)(2)] and 5-dinitro [VO(NO(2) phen)(2)] substitution led to decreased sperm counts and reduced testicular weights. Histopathological examination of testicular sections from VO(Me(2) phen)(2)- and VO(NO(2)-phen)(2)-treated mice revealed a marked inhibition of spermatogenesis and preferential loss of maturing, as well as elongated spermatids. In situ evaluation of seminiferous tubule cross-sections by terminal deoxynucleotidyl transferase-mediated FITC-deoxyuridine triphosphate nick end labeling (TUNEL) and laser scanning confocal microscopy showed characteristic apoptotic germ cells delineating the periphery of the seminiferous tubules. The ability of bis-chelated 4,7-dimethyl- and 5-dinitro-substituted 1,10 phenanthroline complexes of oxovanadium(IV) to induce germ cell apoptosis in vivo may have potential utility in the treatment of human testicular germ cell tumors. PMID- 11481669 TI - Interaction of pyridostigmine and physical stress on antioxidant defense system in skeletal muscle of mice. AB - Pyridostigmine bromide (PB), a reversible anticholinesterase drug, had been used against possible nerve gas exposure during the Persian Gulf War. The Gulf War veterans used PB and they were under physical stress. This study investigated the delayed and interactive effects of pyridostigmine and physical stress on the antioxidant defense system in triceps muscle of mice. Male NIH Swiss mice were divided into four groups and treated as follows: sedentary control; pyridostigmine (1.2 mg kg(-1) p.o.); exercise; and PB plus exercise. Mice were exercised for 10 weeks, but PB was administered daily during the 5th and 6th weeks. Mice were sacrificed 24 h after the last treatments and the triceps muscle was isolated and analyzed. There was a significant increase in total superoxide dismutase (CuZn-SOD + Mn-SOD) activity (141% of control) with PB plus exercise, suggesting that any influx of superoxide anions was scavenged efficiently. The Mn SOD enzyme protein levels were reduced significantly (63% of control) by PB plus exercise. Catalase enzyme protein levels were increased significantly by exercise (132% of control) as well as by PB plus exercise (139% of control). Glutathione levels were increased significantly by exercise alone (123% of control). Pyridostigmine bromide plus exercise significantly increased the malondialdehyde concentration (124% of control) in the triceps muscle, indicating an oxidative stress response of the combination. The data indicate that a combination of PB ingestion and exercise training significantly altered the antioxidant enzyme activities, enzyme protein levels and lipid peroxidation, leading to oxidative injury. Physical stress amplified the delayed effects of PB in the skeletal muscle of mice. PMID- 11481670 TI - Teeth and blood lead levels in Egyptian schoolchildren: relationship to health effects. AB - The objective of this work was to study teeth and blood lead levels in Egyptian schoolchildren and to relate lead levels to sociodemographic and environmental factors, the degree of urbanization and suspected manifestations of possible lead exposure. The study was conducted on 60 children aged 6-12 years: 30 children living in an urban area at Alexandria City and 30 children living in a rural area at Kafr El-Sheikh Province. Both groups are matched for age and gender. Every child was subjected to history taking, clinical examination and IQ measurements. Laboratory investigations included measuring teeth and blood lead levels, haemoglobin, serum iron and total iron binding capacity. The results showed that the mean blood lead level of children in Alexandria was significantly higher than that of the children in Kafr El-Sheikh; also 56.7% and 6.7% of children from Alexandria and Kafr El-Sheikh had a blood lead level of >20 microg dl(-1), with the most frequent symptoms of headache, arthralgia and lack of school interest. The children in Alexandria had significantly lower mean teeth lead and haemoglobin levels than those of the Kafr El-Sheikh group. A history of wrapping sandwiches in newspapers, age and distance between the home and school were significant predictors of lead exposure. These findings support the concept that teeth lead concentration may be a valid addition to the indicators used for assessment of the body burden of environmental lead. In addition, children living in urban areas such as Alexandria need special consideration regarding protection from lead exposure, as well as teeth and blood lead evaluation. PMID- 11481671 TI - Multicopy suppressors of the sly1 temperature-sensitive mutation in the ER-Golgi vesicular transport in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae Sly1 protein is a member of the Sec1/Munc18-family proteins, which are essential for vesicular trafficking, but their exact biological roles are yet to be determined. A temperature-sensitive sly1 mutant arrests the vesicular transport from the ER to Golgi compartments at 37 degrees C. We screened for multicopy suppressor genes that restore the colony formation of the sly1(ts) mutant to discover functionally interacting components. The suppressor genes obtained were classified as: (1) those that encode a multifunctional suppressor, SSD1; (2) heat shock proteins, SSB1 and SSB2; (3) cell surface proteins, WSC1, WSC2 and MID2; (4) ER-Golgi transport proteins, USO1 and BET1; and (5) an as-yet-uncharacterized protein, HSD1 (high-copy suppressor of SLY1 defect 1). By epitope tagging of the gene product, we found that Hsd1 protein is an ER-resident membrane protein. Its overproduction induced enlargement of ER-like membrane structures. PMID- 11481672 TI - High efficiency transformation of Schizosaccharomyces pombe pretreated with thiol compounds by electroporation. AB - A highly efficient method for transformation of the fission yeast Schizosaccharomyces pombe by electroporation has been developed. Significantly higher transformation efficiency was obtained when intact cells grown in SD medium (0.67% Bacto yeast nitrogen base without amino acids, 2% glucose) were pretreated with thiol compounds before an electric pulse was applied to the cells. Among the thiol compounds tested, dithiothreitol (DTT) was the most effective for pretreatment. A high transformation efficiency was obtained when the cells were pretreated with 25 mM DTT at 30 degrees C for 15 min in an osmotically adjusted buffer, since the cells were sensitive to osmotic pressure. It was important to exclude glucose from the DTT pretreatment buffer, as it caused a drastic decrease in efficiency. The optimal cell concentration and amount of DNA during the electric pulse were 1x10(9) cells/ml and 10 ng, respectively. The maximum transformation efficiency, 1.2x10(7) transformants/microg plasmid DNA, was obtained when an electric pulse of 11.0 kV/cm was applied for 5 ms. Furthermore, the high competency of cells pretreated with DTT was maintained by freezing them in a non-permeating cryoprotectant such as sorbitol. PMID- 11481673 TI - Co-consumption of sugars or ethanol and glucose in a Saccharomyces cerevisiae strain deleted in the HXK2 gene. AB - In previous studies it was shown that deletion of the HXK2 gene in Saccharomyces cerevisiae yields a strain that hardly produces ethanol and grows almost exclusively oxidatively in the presence of abundant glucose. This paper reports on physiological studies on the hxk2 deletion strain on mixtures of glucose/sucrose, glucose/galactose, glucose/maltose and glucose/ethanol in aerobic batch cultures. The hxk2 deletion strain co-consumed galactose and sucrose, together with glucose. In addition, co-consumption of glucose and ethanol was observed during the early exponential growth phase. In S.cerevisiae, co-consumption of ethanol and glucose (in the presence of abundant glucose) has never been reported before. The specific respiration rate of the hxk2 deletion strain growing on the glucose/ethanol mixture was 900 micromol.min(-1).(g protein)(-1), which is four to five times higher than that of the hxk2 deletion strain growing oxidatively on glucose, three times higher than its parent growing on ethanol (when respiration is fully derepressed) and is almost 10 times higher than its parent growing on glucose (when respiration is repressed). This indicates that the hxk2 deletion strain has a strongly enhanced oxidative capacity when grown on a mixture of glucose and ethanol. PMID- 11481674 TI - Extensive chromosome translocation in a clinical isolate showing the distinctive carbohydrate assimilation profile from a candidiasis patient. AB - Variation of the electrophoretic karyotype is common among clinical strains of Candida albicans and chromosome translocation is considered one of the causes of karyotypic variation. Such chromosome translocations may be a mechanism to confer phenotypic diversity on the imperfect fungus C. albicans. A clinical strain, TCH23, from a vaginal candidiasis patient shows distinct carbohydrate assimilation profile, serotype B, no chlamydospore formation and an atypical karyotype (Asakura et al., 1991). To examine the taxonomic relationship among C. albicans, Candida dubliniensis and this strain, we sequenced the internal transcribed spacer 1 (ITS1) of nuclear ribosomal DNA. The ITS1 sequence of TCH23 was identical with that of C. albicans but not of C. dubliniensis. Thus, strain TCH23 was classified as a variant of C. albicans with an atypical phenotype. The chromosomal DNAs of this strain were resolved into 13 bands on pulse-field gel electrophoresis (PFGE). Using DNA probes located at or near both ends of each chromosome of C. albicans, we investigated the chromosome organization of this strain. Referring to the SfiI map of C. albicans 1006 (Chu et al., 1993), we found that seven chromosomal DNA bands in strain TCH23 were reciprocal chromosome translocations. One homologue from chromosomes 1, 2 and 6 and both homologues from chromosomes 4 and 7 participated in these events. One translocation product was composed of three SfiI fragments, one each from chromosomes 2, 4 and 7. We deduced the breakpoints of chromosome translocation from the physical map of this strain; between 1J and 1J1, between 2A and 2U, both ends of 4F2, between 6C and 6O and both ends of 7F. PMID- 11481675 TI - The centromere-binding factor Cbf1p from Candida albicans complements the methionine auxotrophic phenotype of Saccharomyces cerevisiae. AB - A gene encoding the centromere binding factor 1 (Cbf1p) of the human pathogenic yeast Candida albicans was cloned and characterized. An open reading-frame was detected which encoded a 223 amino acid protein with a calculated molecular weight of 25.8 kDa and a relative isoelectric point of 5.55. It shares 39% overall amino acid sequence identity with Saccharomyces cerevisiae Cbf1p. We localized the CaCBF1 gene on chromosome 4. Southern analysis indicated that CaCBF1 is probably present as a single copy gene per haploid genome. The CaCBF1 gene under the control of its own promoter was able to complement the methionine auxotrophic growth, the increased mitotic instability of CEN plasmids and the slow growth of a Saccharomyces cerevisiae cbf1Delta mutant strain. PMID- 11481676 TI - The yeast Rvs161 and Rvs167 proteins are involved in secretory vesicles targeting the plasma membrane and in cell integrity. AB - The Rvs161 and Rvs167 proteins are known to play a role in actin cytokeleton organization and endocytosis. Moreover, Rvs167p functionally interacts with the myosin Myo2p. Therefore, we explored the involvement of the Rvs proteins in vesicle traffic and in cell integrity. The rvs mutants accumulate late secretory vesicles at sites of membrane and cell wall construction. They are synthetic lethal with the slt2/mpk1 mutation, which affects the MAP kinase cascade controlled by Pkc1p and is required for cell integrity. The phenotype of the double mutants is close to that described for the pkc1 mutant. Synthetic defects for growth are also observed with mutation in KRE6, a gene coding for a glucan synthase, required for cell wall construction. These data support the idea that the Rvs proteins are involved in the late targeting of vesicles whose cargoes are required for cell wall construction. PMID- 11481677 TI - Identification and characterization of Saccharomyces cerevisiae strains isolated from West African sorghum beer. AB - The occurrence and characterization of yeasts isolated from sorghum beer produced in Ghana and Burkina Faso, West Africa, were investigated. The yeasts involved in the fermentations were found to consist of Saccharomyces spp. almost exclusively. Of the isolates investigated, 45% were identified as Saccharomyces cerevisiae, whereas more than half of the isolates (53%) had physiological properties atypical of S. cerevisiae or any other member of the complex sensu strictu, as they were able to assimilate only glucose, maltose and ethanol as carbon sources. Both ITS-PCR RFLP and PFGE strongly indicated that these isolates were related to S. cerevisiae, regardless of their phenotypic characteristics. Sequencing of the D1/D2 domain of the 26S rDNA confirmed the close relatedness to S. cerevisiae with 0.5% nucleotide differences. The MAL1 and MAL3 loci were found for all isolates as the only recognized MAL loci. Besides, for 40% of the isolates the MAL61 probe hybridized to a position of about 950 kbp, which has not formerly been described as a MAL locus. The results showed that the spontaneous fermentation of West African sorghum beer is dominated by a variety of strains of S.cerevisiae not previously described, among which starter cultures should be selected. PMID- 11481679 TI - Current awareness on yeast. PMID- 11481678 TI - Mutational study of the role of tyrosine-49 in the Saccharomyces cerevisiae xylose reductase. AB - The xyl1 gene encoding xylose reductase was cloned from Saccharomyces cerevisiae and expressed in Escherichia coli. The purified enzyme readily carried out xylose reduction in vitro. It prefers NADPH as the co-enzyme by about 80-fold over NADH. Compared to the native enzyme purified from S. cerevisiae (Kuhn et al., 1995), the recombinant xylose reductase displayed slightly higher (about two-fold) affinities (K(m)) for the substrate (xylose) and co-factor (NADPH), as well as a 3.9-fold faster turnover number (K(cat)) and 7.4-fold greater catalytic efficiency (K(cat)/K(m)). The reason for the apparent discrepancies in kinetic constants between the recombinant and native S. cerevisiae xylose reductases is not known. Replacement of Tyr49 by Phe in the recombinant enzyme led to greater than 98% loss of activity, suggesting that this residue plays a critical role in catalysis. Intrinsic enzyme fluorescence spectroscopic analysis showed that the wild-type and the Y49F variant both bound the co-enzyme NADPH with similar affinity. This supports the view that Tyr49 is involved in interaction with the substrate and not the co-factor during catalysis. PMID- 11481680 TI - Sensory function of basal ganglia. PMID- 11481681 TI - Electrophysiological aids to the diagnosis of psychogenic jerks, spasms, and tremor. PMID- 11481682 TI - Sphincter EMG and differential diagnosis of multiple system atrophy. AB - Multiple system atrophy (MSA) is a degenerative disease manifesting a combination of parkinsonism, cerebellar, pyramidal, and autonomic (including urinary, sexual, and anorectal) dysfunction. It is pathomorphologically defined, but lacks a definitive clinical diagnostic test. Sphincter electromyography (EMG), reflecting Onuf's nucleus degeneration, has been proposed as a helpful test; its value has been reevaluated by a critical review of the literature. In patients with probable MSA, abnormal sphincter EMG, as compared to control subjects, has been found in the majority of patients in all the different forms of the disease in most studies, including patients who, as yet, have no urological or anorectal problems. The prevalence of abnormalities in the early stages of MSA is as yet unclear. Patients with Parkinson's disease (PD) as a rule do not show severe sphincter EMG abnormalities in the early stage of the disease. Anal sphincter EMG abnormalities (abnormal spontaneous activity or motor unit potential changes three standard deviations above valid control data) distinguish MSA from PD in the first 5 years after the onset of symptoms and signs, and from pure autonomic failure, as well as from cerebellar ataxias, if other causes for sphincter denervation have been ruled out. With such criteria, the sensitivity of the method is, however, low. EMG does not distinguish MSA from progressive supranuclear palsy. Future studies should use standardized anal sphincter EMG to better compare results from different centers and precisely define the sensitivity and specificity of the method. PMID- 11481683 TI - Rate of progression in Parkinson's disease: a 6-[18F]fluoro-L-dopa PET study. AB - The aim of this study was to investigate the rate of progression in Parkinson's disease (PD) with 6-[(18)F]fluoro-L-dopa (FDOPA) positron emission tomography (PET). We investigated 21 patients with PD and eight healthy controls. Ten of the patients were de novo at the time of the first PET scan and antiparkinsonian medication was started thereafter, with a favourable response. A FDOPA PET scan was carried out twice at an approximately 5-year interval. The regions of interest were drawn on individual magnetic resonance imaging (MRI) images, matched with the PET images. At the first PET scan, in PD patients the mean k(i)(occ) (x 10(-3) min(-1)) in the anterior putamen was 5.6 +/- 2.7 (mean +/- S.D.; 55% of the control mean) and in the posterior putamen 4.5 +/- 2.4 (45% of the control mean). The k(i)(occ) value for the caudate nucleus was 7.5 +/- 2.1 (x 10(-3) min(-1); 76% of the control mean). The FDOPA uptake declined by the time of the second PET scan and the annual rate of decline was 8.3 +/- 6.3% (P < 0.001) of the baseline mean in the anterior putamen and 10.3 +/- 4.8% (P < 0.001) in the posterior putamen. In the caudate nucleus, FDOPA uptake decreased by 5.9 +/- 5.1% (P < 0.001) of the baseline mean per year. The estimated preclinical period was longest for the posterior putamen being 6.5 years. For the anterior putamen the preclinical period was 4.6 years. In the caudate nucleus, the estimated FDOPA uptake was at normal level at disease onset. In healthy controls, there was no significant decline in FDOPA uptake in any striatal subregion. Our results suggest that the disease process in PD first affects posterior putamen, followed by the anterior putamen and the caudate nucleus, but once started, the absolute rate of decline is the same. In healthy controls, no significant decline in FDOPA was detected. PMID- 11481684 TI - Effect of sleep deprivation on motor performance in patients with Parkinson's disease. AB - Animal research provides evidence that sleep deprivation influences the dopamine system. Knowledge about the effect of sleep deprivation on motor performance in patients with Parkinsons disease is scarce. This study examines the influence of total and partial sleep deprivation compared to normal sleep on motor state and performance in Parkinson's disease. Fifteen nondepressed patients with Parkinson's disease underwent one night of total sleep deprivation (TSD), one night of partial sleep deprivation (PSD) after 3 a.m., and one control night of normal sleep (S), performed in a random, nonconsecutive order. Over a period of 3 hours the following morning, motor evaluations (United Parkinson's Disease Rating Scale, [UPDRS] and tapping rate) were performed before and every 30 minutes after intake of the usual morning dopaminergic drug dose. All patients underwent polysomnography apart from the sleep deprivation protocol. Mean UPDRS motor scores and tapping velocities did not differ significantly after each of the schedules, but a subgroup of four patients improved their motor score after partial sleep deprivation. These data do not confirm previous findings of an overall positive influence of sleep deprivation on motor function in Parkinson's disease. However, the results indicate that different response types to sleep deprivation may exist and that a subgroup of patients could benefit from partial sleep deprivation. PMID- 11481685 TI - Association of REM sleep behavior disorder and neurodegenerative disease may reflect an underlying synucleinopathy. AB - Our objective was to examine whether rapid eye movement (REM) sleep behavior disorder occurs in disproportionally greater frequency in multiple system atrophy (MSA), Parkinson's disease (PD), and dementia with Lewy bodies (DLB), collectively known as the synucleinopathies, compared to other nonsynucleinopathy neurodegenerative disorders. In study 1, we reviewed the clinical records of 398 consecutive patients evaluated at Mayo Clinic Rochester for parkinsonism and/or cognitive impairment. The frequency of suspected and polysomnogram (PSG) confirmed REM sleep behavior disorder (RBD) among subjects with the synucleinopathies MSA, PD, or DLB was compared to the frequency among subjects with the nonsynucleinopathies Alzheimer's disease (AD), frontotemporal dementia (FTD), corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), mild cognitive impairment (MCI), primary progressive aphasia (PPA), and posterior cortical atrophy (PCA). In study 2, we reviewed the clinical records of 360 consecutive patients evaluated at Mayo Clinic Jacksonville for parkinsonism and/or cognitive impairment. The frequency of probable RBD among patients with PD and DLB was compared to the frequency among patients with AD and MCI. In study 3, we reviewed the brain biopsy or postmortem autopsy diagnoses of 23 Mayo Clinic Rochester patients who had been clinically examined for possible RBD and a neurodegenerative disorder. In study 1, patients with MSA, PD, or DLB were more likely to have probable and PSG-confirmed RBD compared to subjects with the nonsynucleinopathies (probable RBD 77/120=64% vs. 7/278=3%, p < 0.01; PSG confirmed RBD 47/120=39% vs. 1/278=0%, p < 0.01). In study 2, patients with PD and DLB were more likely to have probable RBD compared to those with AD and MCI (56% vs. 2%, p < 0.01). In study 3, of the 23 autopsied patients who had been questioned about possible RBD, 10 were clinically diagnosed with RBD. The neuropathologic diagnoses in these 10 included Lewy body disease in nine, and MSA in one. Of the other 13 cases, 12 did not have a history suggesting RBD, and the one case who did had normal electromyographic atonia during REM sleep on PSG and autopsy findings of PSP. Only one of these 13 had a synucleinopathy. The positive predictive values for RBD indicating a synucleinopathy for studies 1-3 were 91.7%, 94.3%, and 100.0%, respectively. Clinically suspected and PSG-proven RBD occurs with disproportionally greater frequency in MSA, PD, and DLB compared to other neurodegenerative disorders. In the setting of degenerative dementia and/or parkinsonism, we hypothesize that RBD is a manifestation of an evolving synucleinopathy. PMID- 11481686 TI - Antiparkinsonian activity and dyskinesia risk of ropinirole and L-DOPA combination therapy in drug naive MPTP-lesioned common marmosets (Callithrix jacchus). AB - De novo administration of long-acting dopamine agonists, such as ropinirole, to patients with Parkinson's disease or to 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-treated subhuman primates produces a lower incidence of dyskinesia than occurs with L-DOPA. This study compares the intensity of dyskinesia produced by combinations of L-DOPA and ropinirole and by these drugs alone, using the MPTP-treated common marmoset model of Parkinson's disease. The objective is to determine the optimum therapeutic strategy for the long-term control of Parkinson's disease with a minimal risk of dyskinesia. MPTP-treated marmosets received either L-DOPA alone, ropinirole alone, or one of two combinations of these drugs (either L-DOPA dominant or ropinirole dominant) daily for 28 days in doses titrated to produce a similar improvement in disability and increase in locomotion. In the group receiving L-DOPA alone, there was a trend for peak dose locomotor activity to increase and the duration of drug effect to decline over the period of the study. L-DOPA alone induced marked dyskinesia over the period of treatment, in contrast to ropinirole which produced a low intensity of involuntary movements. The L-DOPA dominant combination initially produced little dyskinesia, but this became increasingly intense as the study progressed. In contrast, the ropinirole dominant combination produced no greater intensity of dyskinesia than was produced by ropinirole alone. These data suggest that in early Parkinson's disease, the use of ropinirole alone or in combination with a low-dose L-DOPA might delay the induction of dyskinesias while improving motor performance. PMID- 11481687 TI - Neural mechanisms underlying peak-dose dyskinesia induced by levodopa and apomorphine are distinct: evidence from the effects of the alpha(2) adrenoceptor antagonist idazoxan. AB - Dyskinesia, secondary to dopamine replacement therapy, is the major complication of currently available therapies for Parkinson's disease. Alpha(2) adrenoceptor antagonists, such as idazoxan, can significantly reduce levodopa-induced dyskinesia in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned, nonhuman primate model of Parkinson's disease and in human. This action of adrenoceptor antagonists may involve blockade of the actions of noradrenaline synthesised from levodopa. We hypothesise that, because dopamine receptor agonists, such as apomorphine, cannot be metabolised to produce noradrenaline, activation of adrenoceptors may not be involved in dyskinesia produced by such agents. If this were the case, idazoxan would not be expected to reduce apomorphine-induced dyskinesia. MPTP-lesioned marmosets with stable dyskinesia induced by prolonged levodopa therapy were given an acute challenge with apomorphine (0.3 mg/kg subcutaneously) or levodopa (8.0 mg/kg orally), these doses produced equivalent peak-dose dyskinesia. Idazoxan (2.5 mg/kg p.o.), or vehicle, was then administered with either apomorphine or levodopa. Idazoxan abolished levodopa-induced dyskinesia but did not affect apomorphine-induced dyskinesia (P < 0.05 and P > 0.05, respectively, Wilcoxon matched pairs test). Idazoxan also extended the anti-parkinsonian actions of levodopa but did not affect those of apomorphine. The pharmacological characteristics of the neural mechanisms underlying levodopa-induced dyskinesia and apomorphine-induced dyskinesia in parkinsonism thus appear to be distinct, at least with respect to the involvement of alpha(2) adrenoceptors. Specifically, levodopa, but not apomorphine-induced dyskinesia, involves activation of adrenoceptors. This finding may have major implications for understanding dyskinesia and should be borne in mind when designing clinical studies in which levodopa or dopamine receptor agonist challenges are employed to assess potential anti-dyskinetic properties of drugs. PMID- 11481688 TI - Acute effects of bilateral subthalamic nucleus stimulation on clinical and kinematic parameters in Parkinson's disease. AB - After implantation with subthalamic stimulators, nine patients with advanced Parkinson's disease were studied on the task of tracing out, as accurately as possible, the four corners of a square with the dominant hand. The task was performed in four treatment conditions: on stimulation-off medication, off stimulation-off medication, off stimulation-on medication, and on stimulation-on medication. Movement times and peak velocities improved significantly only in the on stimulation-on medication condition compared to off stimulation-off medication. The improvement in clinical parameters with stimulation only (relative to off stimulation off medication) was of borderline significance, while consistent and significant clinical improvement was only obtained with addition of medication (on medication-on stimulation). This study provides quantitative evidence of the effect of subthalamic stimulation on kinematic measures in Parkinson's disease (PD) and suggests that combined treatment (medication and stimulation) is superior to either treatment alone. PMID- 11481689 TI - Corticobasal syndrome with tau pathology. AB - Six cases with a clinical corticobasal syndrome (progressive asymmetric apraxia and parkinsonism unresponsive to levodopa) and tau pathology were selected from 97 brain donors with parkinsonism. Postmortem volumetric measures of regional brain atrophy (compared with age/sex-matched controls) were correlated with clinical features and the degree of underlying cortical and subcortical histopathology. At death, no significant asymmetry of pathology was detected. All cases had prominent bilateral atrophy of the precentral gyrus (reduced by 22-54%) with other cortical regions variably affected. Subcortical atrophy was less severe and variable. Two cases demonstrated widespread atrophy of basal ganglia structures (44-60% atrophy of the internal globus pallidus) and substantial subcortical pathology consistent with a diagnosis of progressive supranuclear palsy (PSP). The remaining four cases had typical pathology of corticobasal degeneration. In all cases, neuronal loss and gliosis corresponded with subcortical atrophy, while the density of cortical swollen neurons correlated with cortical volume loss. Atrophy of the internal globus pallidus was associated with postural instability, while widespread basal ganglia histopathology was found in cases with gaze palsy. This study confirms the involvement of the precentral gyrus in the corticobasal syndrome and highlights the variable underlying pathology in these patients. PMID- 11481690 TI - Comparison of clinical vs. electrophysiological methods of diagnosing of essential tremor. AB - Essential tremor (ET) may be differentiated from normal or enhanced physiological tremor based on a clinical examination or electrophysiological tests such as quantitative computerized tremor analysis. There have been few head to head comparisons of the two methods. Our objective was to estimate diagnostic agreement between these two methods. Cases and controls underwent a clinical evaluation (interview and videotaped examination) and an electrophysiological evaluation (quantitative computerized tremor analysis using accelerometry and electromyography) on the same day, and diagnoses were independently assigned using clinical vs. electrophysiological criteria. Agreement between diagnoses was assessed with a concordance rate and kappa statistic (kappa).Thirty-two (59.3%) of 54 subjects were diagnosed clinically as ET (possible, probable, or definite), compared with 35 (64.8%) of 54 based on tremor analysis. The concordance rate between the two methods of diagnosis was 94.4% (51 of 54). Kappa was 0.88, indicating a level of agreement between diagnoses that was in the "near perfect" range. All of the subjects who received electrophysiological diagnoses of definite ET also received clinical diagnoses of ET. Conversely, all of the subjects who received clinical diagnoses of definite ET also received electrophysiological diagnoses of ET. The agreement between the clinical and electrophysiological diagnosis of ET was substantial, suggesting that study protocols that were to utilize either technique would arrive at similar diagnostic conclusions. In addition, physiological testing can quantify potentially valuable subclinical measurements as well as detect possible additional cases of ET not diagnosed as such during clinical assessments. PMID- 11481691 TI - Somatosensory disinhibition in dystonia. AB - Despite the fact that somatosensory processing is inherently dependent on inhibitory functions, only excitatory aspects of the somatosensory feedback have so far been assessed in dystonic patients. We studied the recovery functions of spinal N13, brainstem P14, parietal N20, P27, and frontal N30 somatosensory evoked potentials (SEPs) after paired median nerve stimulation in 10 patients with dystonia and in 10 normal subjects. The recovery functions were assessed (conditioning stimulus: S1; test stimulus: S2) at interstimuls intervals (ISIs) of 5, 20, and 40 ms. SEPs evoked by S2 were calculated by subtracting the SEPs of the S1 only response from the SEPs of the response to the paired stimuli (S1 + S2), and their amplitudes were compared with those of the control response (S1) at each ISI considered. This ratio, (S2/S1)*100, investigates changes in the excitability of the somatosensory system. No significant difference was found in SEP amplitudes for single stimulus (S1) between dystonic patients and normal subjects. The (S2/S1)*100 ratio at the ISI of 5 ms did not significantly differ between dystonic patients and normal subjects, but at ISIs of 20 and 40 ms, this ratio was significantly higher in patients than in normals for spinal N13 and cortical N20, P27, N30 SEPs. These findings suggest that in dystonia there is an impaired inhibition at spinal and cortical levels of the somatosensory system which would lead to an abnormal sensory assistance to the ongoing motor programs, ultimately resulting in the motor abnormalities present in this disease. PMID- 11481692 TI - Physiological assessment of muscle strength in vitro after direct injection of doxorubicin into rabbit sternocleidomastoid muscle. AB - Doxorubicin chemomyectomy is a potent method for the permanent removal of a muscle or group of muscles after direct local injection, and has been used successfully to treat blepharospasm and hemifacial spasm patients. The efficacy of doxorubicin chemomyectomy on reducing muscle strength after direct injection of doxorubicin into rabbit sternocleidomastoid muscle was tested. One- and 6 month postinjection force assessment was performed in vitro to measure alterations in peak twitch and tetanic force generation, as well as fatigue responses for the treated muscles compared to control. There were significant reductions of both twitch and tetanic peak amplitudes in the doxorubicin-treated muscles. One month after treatment, the decreases in force were greater after 2 mg doxorubicin injections than after 1 mg doxorubicin. While there was a significant reduction in force generation after doxorubicin treatment, fatigue resistances for the doxorubicin-treated muscles were increased compared to the controls. There were significant reductions in muscle mass after doxorubicin treatment, and by 6 months, the myosin heavy chain isoform distribution was similar to normal sternocleidomastoid, except for an increase in slow myosin positive fibers. Doxorubicin chemomyectomy resulted in a significant reduction in functional force generation in the treated sternocleidomastoid muscles. These findings suggest a potential clinical use of doxorubicin chemomyectomy to treat cervical dystonia patients. PMID- 11481693 TI - Home alone: methods to maximize tic expression for objective videotape assessments in Gilles de la Tourette syndrome. AB - Our objective was to test whether at-home videotapes yield enhanced information on tics compared to office-based videotapes and a patient questionnaire on the current anatomical distribution of tics. Ten subjects with Gilles de la Tourette syndrome (age range 8-49 years) who were seen for initial evaluation completed a check list of anatomical areas currently affected with tics, and underwent a videotape examination according to the Rush Videotape Protocol. Each patient/family conducted the same protocol at home at the same time of day within 48 hours. We rated two tapes in random order using the modified published scoring method for the Rush Video-based Tic Rating Scale. Two environments were compared, the doctor's office and at home, with videotapes taken in three conditions: patient engaged in relaxed conversation, patient quietly seated with filmer in the room, and patient quietly seated alone in the room. Data were analyzed using a 2-factor repeated-measures analysis of variance (ANOVA), followed by Wilcoxon signed rank tests. All patients provided excellent quality videotapes that could be scored without difficulty. Environment (office vs. home) and Condition (conversation, quiet with observer, quiet and alone) were both highly significant (P < .0001) and did not interact (P = .54). The highest tic scores for total tic impairment occurred at home with the patient alone (mean score 14.5), and the lowest yield occurred in the office with an observer present (mean score 5.4), the setting closest to the clinical neurological examination. The Home/Alone video segments revealed tics not otherwise seen. Patient questionnaires on body regions recorded more areas than observed in the office videotape, but patients were unaware of several tics captured on the Home/Alone segment. Patients can produce videotapes for objective tic assessments. Because at-home videotapes consistently yield higher tic expressions than in-office films and capture tics that are not appreciated by patients, this methodology is well-suited for enhanced retrieval of objective data on tic expression. PMID- 11481694 TI - Intracerebral recording of readiness potential induced by a complex motor task. AB - While exploring 11 epileptic patients with intracerebral electrodes, we recorded readiness potential (RP) preceding a complex motor task. Multilead depth electrodes were positioned stereotactically into the cortex. In three patients, it was also possible to record RP from the putamen. The movement triggering the recording was the turning of a page in an architectural book. The movement was performed under two conditions: in the first condition, without looking at the pictures on the page (typical self-pacing); and in the second condition, following the inspection of the pictures. There were no significant differences in the appearance of RP under these two conditions, neither in duration nor in amplitude. That could be explained by the fact that "self-paced" does not mean "spontaneous," but covers the internal non-conscious program related to a given task. RP were present in the contralateral primary sensorimotor cortex and the bilateral supplementary motor area (SMA), and in the anterior caudal cingulate cortex. No difference between the cortical topography of RP preceding a simple motor task and the topography of RP occurring in connection with complex movement was observed. PMID- 11481695 TI - Genetic polymorphisms of superoxide dismutase in Parkinson's disease. AB - Oxidative stress reactions may contribute to the pathogenesis of Parkinson's disease (PD). The superoxide dismutases potentially play significant roles in PD by detoxifying superoxide radical. We developed genomic DNA and cDNA-based sequencing assays to identify genetic variants in the copper/zinc superoxide dismutase (SOD1) and manganese superoxide dismutase (SOD2) genes. No genetic variants were detected in the gene encoding SOD1 in DNA from 45 idiopathic PD cases and 49 controls from a population-based case-control study. However, we identified a previously described polymorphism of the mitochondrial targeting sequence consisting of a C47T in exon 2 of SOD2, which results in an alanine to valine substitution. We analyzed this SOD2 variant in DNA from 155 cases and 231 controls from the same study, using an allele-specific fluorogenic 5' nuclease assay, and found no differences in the distributions of allelic frequencies. These results indicate that SOD gene variants do not contribute to PD pathogenesis. PMID- 11481696 TI - Idazoxan, an alpha-2 antagonist, and L-DOPA-induced dyskinesias in patients with Parkinson's disease. AB - Dyskinesia is a frequent and disabling side effect in patients with Parkinson's disease treated with chronic dopa-therapy. Preclinical data in the 1-methyl-4 phenyl-1,2,3,6,-tetrahydropyridine (MPTP) monkey suggest that alpha-2 antagonists may reduce dihydroxyphenylalanine (L-DOPA)-induced dyskinesia. We assessed, in a pilot randomised placebo-controlled study, the effects of single oral doses (10 mg, 20 mg, and 40 mg) of idazoxan, an alpha-2 antagonist, on motor parkinsonian disability and L-DOPA-induced dyskinesia following an acute oral challenge of L DOPA in 18 patients with Parkinson's disease. The severity of L-DOPA-induced dyskinesia improved after 20 mg idazoxan pretreatment, while there was no concommittant deterioration in the antiparkinsonian response to L-DOPA. These results suggest that blocking alpha-2 receptors in patients with Parkinson's disease might improve L-DOPA-induced dyskinesia without the cost of a return of parkinsonian symptomatology. Further studies are required to assess whether this property could have potential therapeutic applications in the long-term management of dyskinetic patients with Parkinson's disease. PMID- 11481697 TI - Cervical dystonia in monozygotic twins: case report and review of the literature. AB - We report on monozygotic male twins of German origin with early onset cervical dystonia. No other family members were affected. Although identical in age of onset and clinical course, there were phenotypic variations. While the second born, more affected twin suffered from a symmetric retrocollis, the twin brother mainly presented with a spasmodic torticollis to the left. Dystonia remained focal in both siblings. A DYT1 gene deletion was excluded. Including our patients, hitherto only four twin pairs with idiopathic dystonia have been reported. Although dystonia in twins can be remarkably uniform in some cases, major differences in terms of disease progression, remissions, and disability may be found. These observations underline the role of inheritance in the pathogenesis of idiopathic dystonia but also indicate that some other factors contribute to the clinical presentation of dystonia. PMID- 11481698 TI - Dystonia in Wilson's disease. AB - The frequency and type of dystonic movements, as well as brain abnormalities, as depicted with magnetic resonance imaging (MRI), which might correlate with dystonia, were studied in 27 consecutive patients with a neurologic form of Wilson's disease (WD) and optimized treatment. Dystonia was found in 10 patients (37%), being generalized in half of them, while two patients had segmental, two patients multifocal dystonia, and one patient bilateral foot dystonia. Dystonia was a presenting sign in four patients and developed later in the course of the disease in six patients, despite the administered therapy for WD. Putamen was the only structure significantly more frequently lesioned in dystonic (80%) in comparison to WD patients without dystonia (24%), suggesting a relation between abnormalities in this brain region and dystonic movements in WD. PMID- 11481699 TI - Familial paroxysmal exercise-induced dyskinesia, epilepsy, and mental retardation in a family with autosomal dominant inheritance. AB - Only few sporadic and familial cases of paroxysmal exercise-induced dyskinesia (PED) have been described in literature. PED associated with familial epilepsy has been rarely reported. We describe a family in which six members in different generations were affected by a long-lasting PED, with childhood onset in five cases. Fasting and stress were also precipitating factors. All the subjects, moreover, showed epileptic seizures during childhood and adolescence. In addition, in all cases a condition of mild mental retardation was also documented, associated in some cases, with irritable and impulsive behaviour. Clinical, neurophysiological, neuroimaging and neuropsychological findings were reported. The homogeneous recurrence of this particular clinical picture in members of three generations emphasised a common genetic basis. In our patients, PED is transmitted as an autosomal dominant trait, with age-dependent penetrance, without evidence of genetic anticipation. The neurophysiological findings suggest a condition of hyperexcitability in the muscular and brain membrane, due to a ion channels disorder. PMID- 11481700 TI - Treatment of restless legs syndrome with the dopamine agonist alpha dihydroergocryptine. AB - An open pilot study with the dopamine agonist alpha-dihydroergocryptine (DHEC) was conducted in 16 patients with idiopathic restless legs syndrome (RLS) over a period of 5 weeks. Following a drug-free interval of 1 week, the patients were treated with daily doses of 10 to 40 mg DHEC. As compared to baseline values, treatment led to a statistically significant reduction of subjective RLS symptoms. Overall complaints at night decreased significantly by 63.9 +/- 38.1% as measured by a visual analogue scale. Detailed evaluation of sensory discomfort, motor restlessness, involuntary movements, as well as sleep quality also showed significant improvement. Side effects were mostly mild and affected mainly the gastrointestinal tract. Five patients needed domperidone for treatment of concomitant nausea. One patient stopped the study due to nausea. In conclusion, the results of this open study suggest a role for DHEC in the treatment of RLS. PMID- 11481701 TI - Development of instruments for abnormal movements: dynamometers, the dynamograph, and tremor recorders. Part 9 of the MDS-Sponsored History of Movement Disorders Exhibit, Barcelona, June 2000. PMID- 11481702 TI - Development of instruments for abnormal movements: postural sway and gait analyses. Part 10 of the MDS-Sponsored History of Movement Disorders Exhibit, Barcelona, June 2000. PMID- 11481703 TI - Seminal figures in the History of Movement Disorders: Hammond, Osler, and Huntington. Part 11 of the MDS-Sponsored History of Movement Disorders Exhibit, Barcelona, June 2000. PMID- 11481704 TI - Focal myopathy as a cause of anterocollis in Parkinsonism. AB - We present a patient with levodopa-responsive Parkinsonism who developed disproportionate anterocollis over a period of 4 to 5 weeks. A neurological evaluation, including EMG and muscle biopsy, demonstrated the presence of a focal myopathy typical of the so-called "dropped head syndrome" or "isolated neck extensor myopathy." Parkinsonian patients presenting with disproportionate anterocollis should be evaluated for focal myopathy, as this may represent one possible cause. PMID- 11481705 TI - Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17): PPND family. A longitudinal videotape demonstration. AB - Frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), characterized by an autosomal dominant inheritance pattern, has recently been recognized as a distinct entity that can display a confusingly broad clinical phenotype. The pallido-ponto-nigral degeneration (PPND) variant is the prototypical example of the parkinsonism-predominant pattern of FTDP-17. A longitudinal videotape demonstration of the clinical progression of this entity in a single individual, along with brief videotape segments from three additional affected individuals, is presented in order to facilitate recognition of this disorder. PMID- 11481706 TI - Truncal and limb apraxia in corticobasal degeneration. AB - We report a patient with probable corticobasal degeneration with prominent truncal apraxia, and review the literature. PMID- 11481707 TI - Case report of pallido-pyramidal disease with supplementary motor area involvement. AB - An C-flumazenil positron emission tomography (PET) study in a patient with pallido-pyramidal disease revealed a marked decrease in benzodiazepine-receptor density in the precentral gyrus cortex and the mesial frontal cortex. We suggest that, in addition to dysfunction of basal ganglia-dependent systems, degeneration of the supplementary motor area could also be involved in the patient's bradykinesia. PMID- 11481708 TI - Severe dysphagia after botulinum toxin injection for cervical dystonia in multiple system atrophy. AB - A 71-year-old woman was treated by botulinum toxin (BTX) type A injections for cervical dystonia related to a multiple system atrophy (MSA). A few days later and persisting for the next 4 months, she developed a severe dysphagia, requiring nasogastric feeding. This implicates cautious use of BTX in a case of MSA. PMID- 11481709 TI - Treatment of neuroleptic malignant syndrome with subcutaneous apomorphine monotherapy. AB - A 20-year-old psychiatric patient receiving haloperidol treatment developed acute onset fever, rigidity, and mental changes. Subcutaneous apomorphine was given alone for treatment. The patient had rapid clinical improvement after the treatment. Serial blood examinations showed decline and subsequent normalization of the creatine phosphokinase levels. PMID- 11481710 TI - Acute paroxysmal dystonia induced by fluoxetine. AB - We report a case of a 32-year-old woman who developed paroxysmal episodes of right hemidystonia 2 days after taking fluoxetine. The attacks subsided 2 days after fluoxetine was withdrawn and did not recur afterwards. To our knowledge, this is the first report of paroxysmal dystonia induced by fluoxetine. PMID- 11481711 TI - Neurostimulation of the ventral intermediate thalamic nucleus in inherited myoclonus-dystonia syndrome. AB - We report on the effects of bilateral neurostimulation of the ventral intermediate thalamic nucleus (VIM) in a patient with medically intractable and progressing inherited myoclonus dystonia syndrome (IMDS). Postoperatively, the patient improved by approximately 80% on the modified version of a myoclonus score without any significant change in the dystonic symptoms. This suggests that neurostimulation of the VIM may be an effective treatment for myoclonus in pharmacologically intractable IMDS. PMID- 11481712 TI - Abolition of postapoplectic hemichorea by Vo-complex thalamotomy: long-term follow-up study. AB - We report a patient with hemichorea following subthalamic hemorrhage. Vo-complex thalamotomy abolished the choreic movements for over 4 years of follow-up. PMID- 11481713 TI - Bilateral pallidal stimulation for idiopathic segmental axial dystonia advanced from Meige syndrome refractory to bilateral thalamotomy. AB - Meige syndrome is an adult-onset dystonic movement disorder that predominantly involves facial muscles, while some patients with this syndrome develop spasmodic dysphonia and dystonia of the neck, trunk, arms, and legs. We report that all dystonic symptoms that had been refractory to both pharmacotherapy and bilateral thalamotomy were markedly alleviated by bilateral pallidal stimulation in a patient with segmental axial dystonia advanced from Meige syndrome. PMID- 11481714 TI - Playing harp, another unusual task-specific dystonia. AB - Herein we report a task-specific dystonia in a 48-year-old woman, with an unusual association between a familial harp-playing dystonia and essential tremor. PMID- 11481715 TI - Case of essential palatal tremor: atypical features and remarkable benefit from botulinum toxin injection. AB - We describe a 21-year-old man with essential palatal tremor. The patient had rhythmic contractions not only of tensor veli palatini but also of facial, lingual, temporalis, pharyngeal, and neck muscles. He had some voluntary control of palatal tremor and ear clicks. He was treated with 5 units of botulinum toxin A (BOTOX) injected into each tensor veli palatini, and had complete resolution of all the symptoms. PMID- 11481716 TI - Modafinil and pramipexole-associated somnolence. PMID- 11481718 TI - Bruxism in Huntington's disease. PMID- 11481720 TI - Rhythmic tremor of the palate and other cranial limb muscles, with cerebellar ataxia: consider Whipple's disease. PMID- 11481721 TI - Ataxia without telangiectasia revisited: update on genetic findings in two brothers with an ataxia-telangiectasia-like disorder. PMID- 11481722 TI - Signing coprolalia and attempts to disguise in a man with prelingual deafness. PMID- 11481723 TI - Effects of reviewing DATATOP videotapes on IQ and MMPI in PSG coordinators and investigators, or, whatever happened to Tony Lang? PMID- 11481724 TI - Vulnerability of rural hospitals to Medicare outpatient payment reform. AB - Because the Balanced Budget Act (BBA) of 1997 requires implementation of a Medicare prospective payment system (PPS) for hospital outpatient services, the authors evaluated the potential impact of outpatient PPS on rural hospitals. Areas examined include: (1) How dependent are rural hospitals on outpatient revenue? (2) Are they more likely than urban hospitals to be vulnerable to payment reform? (3) What types of rural hospitals will be most vulnerable to reform? Using Medicare cost report data, the authors found that small size and government ownership are more common among rural than urban hospitals and are the most important determinants of vulnerability to payment reform. PMID- 11481725 TI - Osteoporosis and hip fractures in the Medicare population, 1992-1996. PMID- 11481726 TI - Assessment of the National Medicare Education Program: supply and demand for information. PMID- 11481727 TI - Medicare proposes 2000 physician fee schedule. PMID- 11481728 TI - New drug regimen helps prevent HIV infection in infants. PMID- 11481729 TI - Physicians weigh in on Medicare reform proposal. PMID- 11481730 TI - New data show continued decline in AIDS deaths. PMID- 11481731 TI - New Jersey expansion of Children's Health Insurance Plan approved. PMID- 11481732 TI - Drug prevention program results found to be modest, slow. PMID- 11481733 TI - Adjusted community rate reforms to promote HMO participation in Medicare+Choice. AB - The authors review the financial regulations imposed on health maintenance organizations (HMOs) that participate in the Medicare+Choice program and identify elements of the regulations that may discourage HMO participation in the program. Modifications of the regulations are proposed that could encourage the participation of HMOs without affording them excessive profit. The modifications include smoothing and bounding profit estimates and authorizing and encouraging expanded use of benefit stabilization funds. PMID- 11481734 TI - Changing patterns of surgical care in the United States, 1980-1995. AB - National inpatient and ambulatory surgery data were combined to examine changes over time in the location and amount of surgical care. The main pattern was a decline in the rate of inpatient operations that was outweighed by growth in ambulatory operations. However, the rate of inpatient operations did not decrease for patients age 65 years or over, despite the growth in ambulatory surgery. Other patterns seen for specific types of operations were the substitution of ambulatory for inpatient operations, increases primarily in the rate of inpatient operations, and decreases in total operations. These patterns have implications for control of health care costs. PMID- 11481735 TI - Capitated payment approaches for Medicaid-financed long-term care services. AB - In the following analysis, the authors examine the capitated payment approaches for long-term care (LTC) services of five programs: the Program of All-Inclusive Care for the Elderly (PACE), the Arizona Long-Term Care System (ALTCS), the Texas STAR + PLUS, the Minnesota Senior Health Option (MSHO), and the Monroe County Continuing Care Networks (CCNs) in New York. The authors describe key aspects in the design of these programs, with an emphasis on Medicaid reimbursement, and discuss differences and commonalities in the approaches taken by the programs in setting capitation rates. PMID- 11481736 TI - Medicaid TEFRA option in Minnesota: implications for patient rights. AB - This research studied a special-needs population under age 18 who had both private insurance and Medicaid coverage through the Tax Equity and Fiscal Responsibility Act of 1982 (TEFRA) option. We found that children with managed care plans, particularly health maintenance organizations (HMOs), tended to incur higher total expenses to TEFRA than children with indemnity plans. Our findings also show that managed care in Minnesota tends to provide the same or marginally better coverage as indemnity plans do for core medical items but much less coverage for ancillary items such as home care, therapies, and durable medical equipment. PMID- 11481737 TI - Hospital, employment, and price indicators for the health care industry: first quarter 1999. AB - This feature presents highlights from statistics on health care utilization, prices, expenses, employment, and work hours, as well as on national economic activity, with brief analysis of these economic indicators. These statistics provide an early indication of changes occurring in the health care sector and within the general economy. Although most data are for the first quarter of 1999, American Hospital Association data (Tables 1 and 2) refer to the third quarter of 1998. PMID- 11481738 TI - An overview: eliminating racial, ethnic, and SES disparities in health care. AB - The papers featured in this issue of the Health Care Financing Review were presented at "Eliminating Racial, Ethnic, and SES Disparities in Health Care: A Research Agenda for the New Millennium." This conference was held on October 15, 1999, in the Washington, DC., area and was co-sponsored by the Health Care Financing Administration (HCFA), the Agency for Healthcare Research and Quality (AHRQ) (formerly the Agency for Health Care Policy and Research), and the Henry J. Kaiser Family Foundation. The conference was undertaken in response to the challenge posed by President Clinton's national goal of eliminating racial and ethnic disparities in six health domains by the year 2010. PMID- 11481739 TI - HCFA's racial and ethnic data: current accuracy and recent improvements. AB - Using linked data from the Medicare Current Beneficiary Survey (MCBS), the authors assessed the accuracy of racial/ethnic classifications in HCFA's enrollment data base (EDB) before and after the 1997 effort to update the EDB. After the update, the sensitivity of the EDB was 97 percent for white persons and 95 percent for black persons, but less than 60 percent for all other categories. The positive predictive value was above 96 percent for white, black, and Hispanic persons, but below 80 percent for all others. There was some improvement in accuracy for white persons and black persons from 1991-1997, and larger improvements for the non-black minorities from 1996-1997. PMID- 11481740 TI - Racial and ethnic differences among Medicare beneficiaries. PMID- 11481741 TI - State children's health insurance program expansion approved. PMID- 11481742 TI - New guideline calls for emphasis on treating tobacco dependence. PMID- 11481743 TI - Disparities in Medicare services: potential causes, plausible explanations, and recommendations. AB - Race/ethnicity and socioeconomic status (SES) are associated with the use of Medicare services. In this article, the author juxtaposes disparities in health outcome measures (including death rates for heart disease, cancer, and stroke) with disparities in the use of elective services expected to improve health, and with disparities in the use of non-elective services associated with poor management of chronic disease. This approach is intended to provide information for judging (a) the reasonableness of the explanations offered for disparities in Medicare utilization and (b) the recommendations made to effect change. PMID- 11481744 TI - A physician's perspective on minority health. AB - In this article, the author discusses her many and varied experiences with health care provision. Her insights into the many dimensions of disparity in health care lead to a set of recommendations for further research. PMID- 11481745 TI - Approaches to eliminating sociocultural disparities in health. AB - The recent flurry of studies documenting the presence of racial, ethnic, and socioeconomic disparities in health care and health have outpaced articles that describe effective strategies to eliminate disparities. Through literature review and informal interviews with research, policy, and program experts, we developed a framework of programs that address disparities through targeting clinicians, patients and communities, and health systems. We found that the lack of technical expertise, resources, and sensitive tools are all common barriers to evaluating programs. To stimulate more effective programs and rigorous evaluations, we describe specialized implementation and evaluation techniques programs can use, and make recommendations for future efforts. PMID- 11481746 TI - Understanding and addressing racial disparities in health care. AB - Racial disparities in medical care should be understood within the context of racial inequities in societal institutions. Systematic discrimination is not the aberrant behavior of a few but is often supported by institutional policies and unconscious bias based on negative stereotypes. Effectively addressing disparities in the quality of care requires improved data systems, increased regulatory vigilance, and new initiatives to appropriately train medical professionals and recruit more providers from disadvantaged minority backgrounds. Identifying and implementing effective strategies to eliminate racial inequities in health status and medical care should be made a national priority. PMID- 11481747 TI - Minority health status in adulthood: the middle years of life. AB - The objective of this article is to describe the racial and ethnic differences in health status during the "middle years" of life. We use data from National Vital Statistics Reports (Hoyert, Kochanek, and Murphy, 1999) to estimate excess mortality among racial and ethnic minority groups for the leading causes of death among adults. Also discussed are the current state of scholarship in minority health and suggestions for future directions for research on racial and ethnic differences in health status. PMID- 11481748 TI - Racial and ethnic differences in hospitalization rates among aged Medicare beneficiaries, 1998. AB - Efforts to study racial variations in access to health care for minorities other than black persons have been hampered by a paucity of data. The Health Care Financing Administration (HCFA) has made efforts in the past few years to enhance the racial codes on the Medicare enrollment files to include Hispanic, Asian American, and Native American designations. This study examines hospitalization rates by these more detailed racial/ethnic groupings. The results show black, Hispanic, and Native American aged beneficiaries compared with white beneficiaries have higher hospitalization rates. Asian American beneficiaries have lower hospitalization rates. Rates of revascularization--coronary artery bypass graft (CABG) and percutaneous transluminal coronary angioplasty (PTCA)- are lower for black, Hispanic, and Native American beneficiaries compared with white beneficiaries, while rates for Asian Americans are similar to rates for white beneficiaries. PMID- 11481749 TI - Medicare risk-adjusted capitation payments: from research to implementation. PMID- 11481750 TI - Implementation of risk adjustment for Medicare. AB - The Health Care Financing Administration (HCFA) implemented risk adjustment for Medicare capitated organizations January 2000. The risk adjustment system used, the Principal Inpatient Diagnostic Cost Group (PIPDCG) method, had to be incorporated into the payment structure mandated by the Balanced Budget Act of 1997 (BBA). This article describes how risk adjustment was integrated into the payment system within the rules of the BBA, and how fee-for-service (FFS) and health maintenance organization (HMO) data are collected and used in the determination of payment. PMID- 11481751 TI - Favorable selection in the Medicare+Choice program: new evidence. AB - Historically, studying the Medicare managed care favorable-selection issue has been difficult because direct data on managed care enrollees have been unavailable. In this study, we analyzed the first year of Balanced Budget Act (BBA)-mandated inpatient encounter data. Based on this comparison of actual managed care and fee-for-service (FFS) beneficiaries, it appears that there are significant differences between these populations. The most striking differences are found in the comparison of average risk factors, indicating a clear bias in the managed care populations toward beneficiaries predicted to be less costly. PMID- 11481752 TI - Risk adjustment for health plans disproportionately enrolling frail Medicare beneficiaries. AB - There is concern about the adequacy of diagnosis-based risk adjusters for paying health plans that disproportionately enroll frail Medicare beneficiaries. The Medicare Current Beneficiary Survey (MCBS) was used to examine the ability of two risk-adjustment models to predict Medicare costs for groups defined by institutional status and difficulty with activities of daily living (ADLs). Both models underpredicted average costs for non-institutionalized frail beneficiaries; however, the models slightly overpredicted expenses for most frail individuals and severely underpredicted for a minority. Further refinements are needed if diagnosis-based models are used to pay plans that disproportionately enroll frail beneficiaries. PMID- 11481753 TI - Modeling Medicare costs of PACE (Program of All-Inclusive Care for the Elderly) populations. AB - Historically, Medicare has paid PACE providers a monthly capitated rate equal to 95 percent of the site's county AAPCC multiplied by a PACE-specific frailty adjuster of 2.39. The Balanced Budget Act of 1997 makes PACE a permanent provider category and mandates that future Medicare payments be based upon the rate structure of the Medicare+Choice payment system, adjusted for the comparative frailty of PACE enrollees and other factors deemed to be appropriate by the Secretary of Health and Human Services. This study revisits the calculation of the PACE frailty adjuster and explores the effect of risk adjustment on that frailty adjuster. PMID- 11481754 TI - Health-related quality of life predictors of survival and hospital utilization. AB - The health-related quality of life (HRQOL) of 103 end-stage renal disease (ESRD) patients on hemodialysis was studied for prediction of 1-year survival and hospital days in the context of other predictors. Higher HRQOL physical functioning, higher provider-reported functional performance, fewer private religious activities, living with family, black race, and having a diagnosis of hypertension predicted survival. Lower HRQOL energy, higher pain, and not living with family predicted more hospital days. Patients living with family reported more social support and better HRQOL general health, emotional well-being, social health, and quality of social interactions than other patients. PMID- 11481755 TI - Impact of expanding SSI on Medicaid expenditures of disabled children. AB - Supplemental Security Income (SSI) expansions for disabled children in the early 1990s provoked criticism that eligibility criteria were too lax and motivated the subsequent retraction of benefits for many children. However, little evidence exists on whether the clinical needs of SSI children declined during this period. The authors used Medicaid data to examine changes in average expenditures between 1989 and 1992, using an Aid to Families with Dependent Children (AFDC) comparison group to control for confounding time trends (e.g., in access). Results showed declines in average expenditures in Georgia and Tennessee but increases in California and Michigan, which are thought to have started with more liberal eligibility policies. PMID- 11481756 TI - Residential care supply, nursing home licensing, and case mix in four states. AB - Simulation analyses quantify admission and continuing physical and cognitive impairment patient case-mix changes under two scenarios: with increases in residential care supply and with all nursing homes licensed only as skilled care facilities. Findings raise caution about the assumed interplay between residential care supply and nursing home use. The proportion of nursing home patients with only physical and cognitive impairment likely to be affected by current and emerging long-term care (LTC) policy was well under 25 percent of the nursing home population in each of the four study States. States varied in LTC supply and utilization controls. PMID- 11481757 TI - Hospital, employment, and price indicators for the health care industry: third quarter 1999. AB - This feature presents highlights from statistics on health care utilization, prices, expenses, employment, and work hours, as well as on national economic activity, with brief analysis of these economic indicators. These statistics provide an early indication of changes occurring in the health care sector and within the general economy. Although most statistics include data for third quarter of 1999, American Hospital Association (AHA) data are through the third quarter of 1998. PMID- 11481758 TI - Risk adjustment and the health of the Medicare HMO population. PMID- 11481759 TI - Maine Medicaid early HIV treatment plan approved. PMID- 11481760 TI - New insurance option to cover breast and cervical cancer. PMID- 11481761 TI - Help is available to pay Medicare Part B premium. PMID- 11481762 TI - Pennsylvania expansion of State Children's Health Insurance Program approved. PMID- 11481763 TI - Government initiative seeks to determine environmental causes of disease. PMID- 11481764 TI - Office-based Medicaid managed care physicians serve few Medicaid patients. PMID- 11481765 TI - Children's Health Insurance Program now reaching two million. PMID- 11481766 TI - Health care spending growth rates stay low. PMID- 11481767 TI - Improving health-based payment for Medicaid beneficiaries: CDPS. AB - This article describes the Chronic Illness and Disability Payment System (CDPS), a diagnostic classification system that Medicaid programs can use to make health based capitated payments for TANF and disabled Medicaid beneficiaries. The authors describe the diversity of diagnoses and different burdens of illness among disabled and AFDC Medicaid beneficiaries. Claims from seven States are analyzed, and payment weights are provided that States can use when adjusting HMO payments. The authors also compare the taxonomy and statistical performance of CDPS to other leading diagnostic classification systems and find that the new model performs better in a number of respects. PMID- 11481768 TI - A clinically detailed risk information system for cost. AB - The authors discuss a system that describes the resources needed to treat different subgroups of the population under age 65, based on burden of disease. It is based on 173 conditions, each with up to 3 severity levels, and contains models that combine prospective diagnoses with retrospectively determined elements. We used data from four different payers and standardized the cost of most services. Analyses showed that the models are replicable, are reasonably accurate, explain costs across payers, and reduce rewards for biased selection. A prospective model with additional payments for birth episodes and for serious problems in newborns would be an effective risk adjuster for Medicaid programs. PMID- 11481769 TI - Using diagnoses to describe populations and predict costs. AB - The Diagnostic Cost Group Hierarchical Condition Category (DCG/HCC) payment models summarize the health care problems and predict the future health care costs of populations. These models use the diagnoses generated during patient encounters with the medical delivery system to infer which medical problems are present. Patient demographics and diagnostic profiles are, in turn, used to predict costs. We describe the logic, structure, coefficients and performance of DCG/HCC models, as developed and validated on three important data bases (privately insured, Medicaid, and Medicare) with more than 1 million people each. PMID- 11481770 TI - Principal inpatient diagnostic cost group model for Medicare risk adjustment. AB - The Balanced Budget Act (BBA) of 1997 required HCFA to implement health-status based risk adjustment for Medicare capitation payments for managed care plans by January 1, 2000. In support of this mandate, HCFA has been collecting inpatient encounter data from health plans since 1997. These data include diagnoses and other information that can be used to identify chronic medical problems that contribute to higher costs, so that health plans can be paid more when they care for sicker patients. In this article, the authors describe the risk-adjustment model HCFA is implementing in the year 2000, known as the Principal Inpatient Diagnostic Cost Group (PIPDCG) model. PMID- 11481771 TI - Measuring health spending. Overview. PMID- 11481772 TI - Medical expenditures for major diseases, 1995. AB - This article distributes the Health Care Financing Administration's (HCFA) estimates of 1995 personal health care expenditures (PHCE) according to sex, age, and diagnosis for each type of health care service. Aggregate and per capita expenditures are reported for 18 broad categories of disease classified according to the International Classification of Diseases (ICD-9-CM). Special emphasis is given to expenditures for persons age 65 or over, the segment of the population with the highest expenditures. These results show how the relative importance of medical conditions and type of health services differs between the sexes and changes with increasing age. PMID- 11481773 TI - Matching health policy with data: data and analytic requirements for federal policymakers. AB - The health data and statistical needs of our health care system continue to grow. Though we are expected to spend approximately $1.4 trillion on health care next year, we know little about where the dollars are spent and what they are purchasing. Our national health statistics are currently collected through a patchwork of claims data and survey data. These data are collected periodically, are often out of date, and do not contain several key data elements critical for serious evaluation of the performance of our health care system. Failure to collect more timely and comprehensive data will undermine ongoing efforts for controlling the growth in costs and improving quality. PMID- 11481774 TI - National health expenditures, 1998. AB - In 1998, national health care expenditures reached $1.1 trillion, an increase of 5.6 percent from the previous year. This marked the fifth consecutive year of spending growth under 6 percent. Underlying the stability of the overall growth, major changes began taking place within the Nation's health care system. Public payers felt the initial effects of the Balanced Budget Act of 1997 (BBA), and private payers experienced increased health care costs and increased premium growth. PMID- 11481775 TI - National health projections through 2008. AB - Health spending is expected to resume its rise as a share of gross domestic product (GDP) in the projection period, following 6 years of near stability, increasing from 13.5 percent in 1997 to an estimated 16.2 percent by 2008. This implies an approximate doubling of health spending, from $1.1 trillion in 1997 to $2.2 trillion by 2008. We anticipate a reversal in recent patterns of growth in public and private health spending, with private spending expected to accelerate while Medicare spending slows in response to the implementation of the Balanced Budget Act (BBA) of 1997. PMID- 11481776 TI - Hospital, employment, and price indicators for the health care industry: second quarter 1999. AB - This feature presents highlights from statistics on health care utilization, prices, expenses, employment, and work hours, as well as on national economic activity, with brief analysis of these economic indicators. These statistics provide an early indication of changes occurring in the health care sector and within the general economy. Although most statistics include data for the second quarter of 1999, American Hospital Association data are through the third quarter of 1998. PMID- 11481777 TI - State health expenditure accounts: purposes, priorities, and procedures. AB - This article reports on the State Health Expenditure Account (SHEA) project which developed procedures States can use in tracking their health care expenditures. The purposes, priorities, and concepts of SHEAs were designed to meet the needs of State policymakers. The resulting methods are discussed and illustrated using calculations of SHEAs for California. Contrasts with the National Health Expenditure (NHE) framework are provided. Recommendations for cooperation between the Health Care Financing Administration (HCFA) and the States that would facilitate the adoption and estimation of SHEAs are offered. Details of the methods used for the California estimates can be found in the Technical Note of this article. PMID- 11481778 TI - Health expenditures for Medicare beneficiaries. PMID- 11481779 TI - Information needs and preferences of the general Medicare population. PMID- 11481780 TI - New tobacco industry documents Website unveiled. PMID- 11481781 TI - California expansion of children's health insurance program approved. PMID- 11481782 TI - Surgeon General reports on mental health revolution. PMID- 11481783 TI - Guidance available on communicating with speech/hearing-impaired elderly. PMID- 11481784 TI - National standards proposed to protect patients' personal medical records. PMID- 11481785 TI - What can the U.S. learn from national health accounting elsewhere? AB - The United States is typically seen as an outlier in health spending when compared with other advanced nations. Recent improvements in health accounting in lower- and middle-income countries suggest some common features with the high and pluralistic spending in the United States. The author discusses recent developments and findings in health accounting outside the Organization for Economic Cooperation and Development (OECD) and their relevance for the United States. He argues that we should expect more fruitful exchanges in the future. PMID- 11481786 TI - Future directions for the national health accounts. AB - Over the past 15 years, the Health Care Financing Administration (HCFA) has engaged in ongoing efforts to improve the methodology and data collection processes used to develop the national health accounts (NHA) estimates of national health expenditures (NHE). In March 1998, HCFA initiated a third conference to explore possible improvements or useful extensions to the current NHA projects. This article summarizes the issues discussed at the conference, provides an overview of three commissioned papers on future directions for the NHA that were presented, and summarizes suggestions made by participants regarding future directions for the accounts. PMID- 11481787 TI - State health expenditure accounts: Minnesota's perspective. AB - Minnesota's approach to the development and use of State health expenditure accounts (SHEAs) was developed to assist State policymakers with decisions regarding health care reform. The accounts are based on an annual survey of third party payers and summary Medicaid and Medicare data. Summary data are presented along with a discussion of data collection methodology, estimation, and dissemination. Minnesota's experience demonstrates that the ability of States to conduct detailed analysis of health care spending and to use these estimates to change State policy, inform national policy debate, conduct impact analysis, educate policymakers, and monitor market trends. PMID- 11481788 TI - State-level variation in Medicare spending. AB - Theoretically, Medicare provides one standard benefit package to all enrollees. But because of State-level variations in populations, service supply, and local practice patterns, national policy changes may have unequal impacts on access and service utilization. Across-the-board policy changes may create hardships in one area while appropriately discouraging use in another area. In this article, the authors describe State-level variations in Medicare enrollees, their insurance coverage, 1995 Medicare and beneficiary spending patterns in aggregate, per capita, and by service, and certain spending patterns for dually eligible beneficiaries. These data are useful for considering the State-level effects of payment reform. PMID- 11481789 TI - Health expenditure trends in OECD countries, 1970-1997. AB - This article provides an overview of current trends in health expenditures in 29 OECD countries and recent revisions of OECD health accounts. U.S. health expenditures are compared with those of other OECD countries. The interactions of cost-containment measures with changes in the public-private mix of financing and in the composition of health care spending are discussed. PMID- 11481790 TI - TDI pledges to correct prompt payment problems. PMID- 11481791 TI - The doctor as a witness: medicine on trial. PMID- 11481792 TI - MedBytes. PMID- 11481793 TI - The battle continues. Congress still wrestling with competing patients' rights bills. PMID- 11481794 TI - Guidelines or physicians' judgment? PMID- 11481795 TI - Crossing a new frontier. One physician's vision of border health is now a binational reality. PMID- 11481796 TI - Delegated networks. Their relationship with insurers causes concern. PMID- 11481797 TI - Reinventing the library. Association seeks to improve TMA library services, efficiency. PMID- 11481798 TI - Do "conditions contributing to the death" clarify the predictors of infant mortality in Texas? AB - Predictors of infant mortality are usually based on the underlying causes reported on infant death certificates. Death certificates also contain space for "Other significant conditions contributing to the death but not resulting in the underlying cause." We investigated whether these "contributing conditions" might clarify the predictors of infant death in Texas or interventions that might reduce deaths from the underlying causes. A "multiple cause of death" file converted the underlying causes and contributing conditions associated with each infant death in 1996 into International Classification of Diseases codes. We ranked the major underlying causes, the conditions contributing to those causes, the conditions contributing to all infant deaths, and the effect of combining the underlying causes with contributing conditions to determine the main predictors of infant mortality. We found that extreme immaturity was the most common condition, causing or contributing to 604 of the 2081 deaths in 1996. We conclude that the underlying causes provide a misleading perception of the major predictors of infant death in Texas; combining them with the conditions contributing to those deaths provides a clearer picture. Given the powerful predictive value of preterm birth for infant mortality, the limited current ability to prevent preterm birth, and the cost of its complications, expanded efforts to improve understanding of its causal processes and develop preventive strategies should be top national and state priorities. PMID- 11481800 TI - Voice of experience. PMID- 11481799 TI - The importance of infant mortality statistics. PMID- 11481801 TI - Thinking about the unthinkable. Where to start planning for terrorism incidents. PMID- 11481802 TI - New codes blooming. Spring means pleasant weather and the NFPA world congress. PMID- 11481803 TI - Sitting pretty. Taking the risks out of cleaning upholstered furniture. PMID- 11481804 TI - First person. Uniform values. PMID- 11481805 TI - Nurse consultants. Room at the top. AB - The role of nurse consultant needs to be embedded in trust culture if it is to achieve its potential. The process of defining key competencies can enhance the organisation's ownership. The post needs to be evaluated at local level. PMID- 11481806 TI - Radiology. Image problem. AB - The importance of radiology departments is growing in all acute hospitals. A review of skill-mix could reduce pressure on radiology departments. The role of radiographers trained in sonography should be extended. PMID- 11481807 TI - Open space. Pulling back the screens. PMID- 11481808 TI - Midwest cap rates reflect continued upward movement. PMID- 11481809 TI - Capitated groups learning to manage risk, report shows. PMID- 11481810 TI - HMO enrollment declines, but capitation holding its own. PMID- 11481811 TI - New tool helps benchmark, negotiate risk contracts, and test capitation scenarios. PMID- 11481812 TI - Widely different referral rates have little effect on outcomes. PMID- 11481813 TI - Digital doctoring 2002. PMID- 11481814 TI - Stats & Facts. Prostate-specific antigen screening. PMID- 11481815 TI - Changing health care: conformists or trailblazers? PMID- 11481816 TI - Complex treatment options for rheumatoid arthritis. PMID- 11481817 TI - Coverage inequalities of new therapies for rheumatoid arthritis in a managed care setting. AB - Rheumatoid arthritis (RA) is a progressive disease that leads to functional disability and substantial medical costs. Early treatment with disease-modifying antirheumatic drugs (DMARDs) has been shown to inhibit the progression of RA, with accompanying improvements in functional outcome and long-term medical costs. Biologic response modifiers (BRMs) are a new class of therapeutic agents for RA that offer a more favorable side-effect profile than traditional DMARDs. One BRM, the self-injectable medication etanercept, has demonstrated great potential for improving the long-term prognosis of patients with RA. Unfortunately, patients' access to treatment with etanercept is sometimes limited because of the structure of health-care coverage for self-injectable drugs. PMID- 11481818 TI - Pharmaceutical price controls at the state level. PMID- 11481819 TI - Mild-to-moderate uncomplicated hypertension: further analysis of a cost effectiveness study of five drugs. AB - A cost-effectiveness model was designed to explore the effect of adding a new angiotensin-II inhibitor, telmisartan, to the therapeutic options for treating mild-to-moderate uncomplicated hypertension. Incorporating the cost of drugs, physician visits, and adverse-event treatments, the model concluded that availability of telmisartan on formulary may shorten the mean time and costs to control. The stability of the initial findings over a range of sensitivity analyses lends credence to the model conclusions that availability of telmisartan on formulary improves the therapeutic options of care for hypertension. PMID- 11481820 TI - [Assessment of physical activities from functional physical fitness tests research for patients with chronic obstructive pulmonary disease]. AB - The purpose of this study was to determine whether the level of physical activity (as measured by 12 functional physical fitness tests) differed between patients with chronic obstructive pulmonary disease (COPD) and normal subjects. Fifty male patients with COPD (70.4 +/- 7.1 yr) and 193 age-matched normal subjects (71.2 +/ 5.2 yr) participated in the study. We administered twelve functional physical fitness tests that were designed to represent muscle strength, muscular endurance, reaction, flexibility, balance, regulation and manipulation. Cardiorespiratory fitness was measured by an incremental, symptom-limited exercise test to the maximal tolerable level on a cycle ergometer, and by the 6 min walk distance. Further, we evaluated what the relationships were between cardiorespiratory fitness and physical activities. The results showed that there were significant differences between COPD and normal groups in cardiorespiratory fitness [peak oxygen uptake (15.7 +/- 4.8 ml/kg/min vs. 23.9 +/- 6.5 ml/kg/min, p < 0.05), peak load (77.9 +/- 33.7 watts vs. 110.3 +/- 37.9 watts, p < 0.05) and 6 min walk distance (466.4 +/- 106.3 m vs. 540.4 +/- 78.3 m, p < 0.05)] and muscular endurance [arm curl (22.6 +/- 5.0 rep/30 s vs. 25.2 +/- 6.0 rep/30 s, p < 0.05), Keeping a half-squat position (32.6 +/- 16.4 s vs. 42.8 +/- 17.7 s, p < 0.05)] and regulation (27.2 +/- 9.4 s vs. 23.3 +/- 5.8 s, p < 0.05) in functional physical fitness test. There were significant correlations between cardiorespiratory fitness, and muscular endurance and regulation. Furthermore, there were significant correlations between FEV1.0 and FEV1.0% pred, and regulation. Therefore, it is important that, besides exercise prescriptions based on the criteria of symptom-limited exercise tests, development of exercise programs based on the level of physical activities may be necessary for patients with COPD. PMID- 11481821 TI - [Clinical significance of sialyl SSEA-1 (SLX) in patients with idiopathic interstitial pneumonia]. AB - Serum SLX levels were measured in 29 patients with idiopathic interstitial pneumonia (IIP) to evaluate its clinical significance. Serum SLX had positive correlations with the BALF neutrophil ratio but not with the severity or the disease activity in patients with IIP, and bronchoalveolar lavage fluid (BALF) SLX had positive correlations with the BALF neutrophil count. Epithelial lining fluid (ELF) SLX levels showed positive correlations with serum SLX, but were much higher. These results suggest that increases of serum SLX may reflect increases of SLX in the lung tissues. Thus, we speculate that increases of the serum SLX level may represent increases of the BALF neutrophil count. Patients with higher SLX showed poor therapeutic responses and poor prognoses in comparison with those with normal SLX for the reason that serum SLX level represents BALF neutrophil level. PMID- 11481822 TI - [Relation between neck accessory inspiratory muscle electromyographic activity and lung volume]. AB - Years ago it was reported that, as lung volume increased, there was a corresponding increase in scalene EMG activity (Raper et al. J Appl Physiol 21: 497-502, 1966). Otherwise, the relationship between changing lung volume and the EMG of the respiratory muscles has not been defined. We therefore inserted fine wire electrodes into the scalene (SCLN), sternocleidomastoid (STERNO), and trapezius (TRAPEZ) muscles in 6 healthy subjects under direct vision using high resolution ultrasound. Maximum EMG activity (EMGmax) was obtained for each muscle by a variety of respiratory and postural maneuvers. Then, in the standing posture, air flow, raw and moving average EMG signals were sampled and input to a computer during quasi-static inspiration from functional residual capacity (FRC) to total lung capacity (TLC). We found that the relationship between EMG and lung volume for SCLN, but not for STERNO or TRAPEZ, was expressed by exponential curves. The onset of SCLN, STERNO and TRAPEZ EMG occurred at 13.3 +/- 7.4 (mean +/- SE), 67.8 +/- 14.6, and 89.2 +/- 3.9% of inspiratory capacity. The EMG of SCLN, STERNO, TRAPEZ, reached 85.7 +/- 2.6, 60.7 +/- 8.6, and 11.8 +/- 5.2% of EMGmax, respectively, at TLC. We conclude that: 1) SCLN is the most, and TRAPEZ the least, active neck accessory inspiratory muscle, while STERNO is intermediate, and 2) there is a lung volume dependency of the neck accessory muscle EMG activity. PMID- 11481823 TI - [Two cases of pulmonary disease with perinuclear anti-neutrophil cytoplasmic antibody]. AB - We encountered two cases of perinuclear anti-neutrophil cytoplasmic antibody (p ANCA). The first was a case of idiopathic interstitial pneumonia diagnosed in a 73-year-old man since 1998. He was admitted to our hospital because of renal failure and anemia. The serum level of p-ANCA on admission was 264 EU, and specimens obtained by percutaneous renal biopsy showed crescentic glomerulonephritis and vasculitis due to p-ANCA. He was treated with prednisolone pulse therapy and prednisolone (PSL), however interstitial pneumonia occurred during PSL tapering. We treated him for pulmonary fibrosis with plasmapheresis, methylprednisolone (mPSL) and cyclophosphamide (CPA), which suppressed the progress of the interstitial pneumonia. The second case was one of massive pulmonary hemorrhage in a 68-year-old man who was admitted to our hospital. Physical examination revealed anemia: the laboratory data, renal failure; and the serum level of p-ANCA was elevated to 611 EU. The specimens obtained by percutaneous renal biopsy showed crescentic glomerulonephritis and vasculitis. The renal failure was not improved by PSL, but, together with the inflammation, responded to the combination of PSL and CPA. However, both patients died of serious infection. They were regarded as compromised patients because of the therapy mentioned above. No standard therapy has been established against p-ANCA positive pulmonary disease with renal failure. The treatment should control the progression of interstitial pneumonitis and pulmonary hemorrhage. It is important to consider the possibility of serious infection. PMID- 11481824 TI - [Two cases of pneumothorax using the thoracic vent (TV) on an outpatient basis]. AB - In case 1, a 73-year-old man developed dyspnea on exertion while travelling abroad. The symptom continued until the patient returned to Japan 5 days after onset and visited our clinic on the next day. SaO2 was 93% and chest radiography demonstrated pneumothorax with 75% collapse of the right lung. Insertion of a TV successfully prevented air leakage stopped within 2 days. The TV was removed after the lung collapse disappeared, as shown on the chest radiograph taken 4 days after its insertion. Case 2 occurred in a 62-year-old woman with underlying idiopathic interstitial pneumonitis, who had had dyspnea on exertion for approximately 5 years. The patient visited our clinic because the dyspnea on exertion became abruptly exacerbated. A TV was inserted after an SaO2 of 82% was measured and the chest radiograph revealed pneumothorax with a 50% collapse of the right lung. Air leakage stopped after 5 days, and the TV was removed on the 8th day after insertion. We concluded that TV is very useful for improving the QOL of patients and enables outpatient management of pneumothorax, resulting in a reduction in the cost of treatment. PMID- 11481825 TI - [Atelectasis of the right lower lobe in association with bronchiolitis obliterans organizing pneumonia]. AB - A 58-year-old woman with underlying rheumatoid arthritis was admitted to the hospital because of a dry cough and the presence of an abnormal shadow in the right lower lung field. Consolidation and volume loss in the right lower lobe with air bronchogram were recognized on a chest tomogram. Bronchofiberscopic examination disclosed neither stenosis nor tumors in the large bronchi. Organizing pneumonia was recognized pathologically in transbronchial lung biopsy (TBLB) specimens. The volume of the right lower lobe decreased rapidly, and new infiltration shadows appeared in the right upper and middle lobes. Another bronchofiberscope examination revealed organizing pneumonia, and macrophage infiltrations were seen in the alveoli on histopathological examination of the TBLB specimens. The diagnosis of RA-associated BOOP was made on the basis of agreement of the expansion of the shadows on chest radiographs, the RA symptoms and the RA factor. The patient was treated with prednisolone, and the clinical course was satisfactory, with no recurrence. This case was of interest because BOOP inducing lobar cicatricial atelectasis is very rare. PMID- 11481826 TI - [A case of mucoid impaction of bronchi (MIB) due to Schizophyllum commune]. AB - A 51-year-old woman was admitted because of a productive cough. Atelectasis of the left upper lobe and eosinophilia were noted. The atelectasis was resolved after bronchoscopic removal of the mucous plug. Pathologically, it consisted of eosinophil accumulation with hyphae. Cultures of sputum samples revealed Schizophyllum commune. We diagnosed this case as mucoid impaction of bronchi due to S. commune. Eight months later, a productive cough developed again. Chest radiography showed atelectasis of the right middle lobe. Cultures of bronchial washings yielded S. commune and Aspergillus niger. The atelectasis was resolved by four months of administration of itraconazole (200 mg daily). PMID- 11481827 TI - [A case of chronic interstitial pneumonia with transient acute excerbation induced by dust exposure and spontaneous remission by the admission to hospital]. AB - A 54-year-old man was admitted to our hospital with a chief complaint of dyspnea. He has been a plasterer with exposure to dust for 35 years since the age of 19 years. Initially, nonspecific interstitial pneumonia was diagnosed because of findings of ground-glass opacity without honeycombing on chest radiography and CT scanning. Restrictive and diffusional dysfunction of the lung was observed through pulmonary function testing. Videoscope-assisted thoracic surgery revealed pleural plaques and a lung histology showing usual interstitial pneumonia (UIP). Asbestos bodies and peri-bronchial and vascular dust deposition were detected by microscope. This patient recovered without any medication, with verification from the pulmonary function test results, chest radiographs and CT scanning results. The diagnosis was chronic interstitial pneumonia with transient excerbation. The cause of these pulmonary changes was thought to be dust exposure. Therefore, since dust exposure was avoided after the patient's admission to hospital, dust (including asbestos) exposure may be an important factor in the acute excerbation. PMID- 11481828 TI - [A case of micronodular pneumocyte hyperplasia, lymphangiomyomatosis associated with tuberous sclerosis]. AB - We report a case of tuberous sclerosis (TS) associated with renal angiolipomas, subungual fibroma, adenoma sebaceum, pulmonary lymphangiomyomatosis (LAM), and micronodular pneumocyte hyperplasia (MNPH). A CT scan of the chest demonstrated multiple minute nodules present throughout both lung fields. Histologically, the nodules were well demarcated, and were composed of thickened, fibrotic, alveolar septa lined by pleomorphic, type II pneumocytes. This type of MNPH is extremely rare, and appears to be a hamartomatous proliferation occurring in patients with TS. PMID- 11481829 TI - [A recurrent case of idiopathic interstitial pneumonia histologically diagnosed as nonspecific interstitial pneumonia group 1]. AB - A 59-year-old man was admitted to our hospital because of dyspnea. Chest radiography showed infiltration and consolidation in both lung fields. He was clinically diagnosed as having idiopathic interstitial pneumonia (IIP), and histological examination of a thoracoscopic lung biopsy specimen showed nonspecific interstitial pneumonia (NSIP) group 1. Corticosteroid therapy had previously been effective, but about 1 year later the disease recurred. The patient's clinical condition was very similar to the first episode, and is improving in response to the same treatment again. Although it is generally accepted that patients with IIP diagnosed histologically as NSIP have a good prognosis, it should be remembered that recurrence is possible, even in patients with group 1. PMID- 11481830 TI - [Aortitis syndrome associated with ulcerative colitis, preceded by pulmonary infarction involvement]. AB - A 21-year-old woman with a 6-year history of ulcerative colitis admitted to our hospital with chest pain, cough and fever of unknown origin in August 1998. On admission, laboratory data showed positive inflammatory signs. A chest radiograph and chest computed tomogram (CT) revealed nodular shadows in the right upper lung field. Fifty days after admission, hypertension developed and a bruit was audible in the neck and the upper abdomen. Digital subtraction angiography showed stenosis in carotid, renal and right upper pulmonary arteries. On the basis of these results, a diagnosis of aortitis syndrome was made. Moreover, these findings indicated pulmonary infarction in the right upper lobe due to aortitis syndrome. Aortitis syndrome preceded by pulmonary infarction involvement is very rare. Autoimmune disorders may have been involved in this case because of the association with ulcerative colitis. PMID- 11481831 TI - [Cave-associated acute pulmonary histoplasmosis in two Japanese returning from Mexico]. AB - We report cases of cave-associated acute pulmonary histoplasmosis that occurred in two Japanese returning from Yucatan, Mexico. Patient 1 is a 35-year-old woman who worked in a bat-infested cave in Mexico for about 3 weeks. Almost all her colleagues had developed cough, fever and headache after 5 days in the cave. She was asymptomatic but her chest radiograph showed multiple nodules 5-10 mm in diameter throughout both lungs. The histoplasmal mycelial-phase complement fixation titer on admission was 1:16, and 1:128 one month later. Patient 2, the 53-year-old husband of patient 1, was also infected by histoplasma in the cave, and similar nodules were observed in his chest radiograph. His histoplasmal complement fixation titer did not elevate. To date, about 20 cases of pulmonary histoplasmosis have been reported in Japan. The number of imported mycoses in Japan is increasing, and histoplasmosis should be considered in the differential diagnosis of pulmonary nodules in travelers returning from endemic areas. PMID- 11481832 TI - [New serum markers to monitor treatment of acute exacerbation of interstitial lung disease]. AB - A 58-year-old woman was admitted to our hospital because of recurrent fever, severe cough and sputum. Chest radiological examinations showed diffuse reticulonodular opacities in both lung fields. Interstitial pneumonia with probable polymyositis was diagnosed. Serum surfactant protein (SP)-A, SP-D and KL 6, which are new interstitial lung disease markers, showed values significantly higher than cutoff levels. The markers increased more in parallel with the rapid development of respiratory insufficiency, CPK level, myalgia and proximal muscle weakness. Treatment with a high dose of corticosteroid and the following gradual decrease over 8 months led to clinical and radiological improvement, with normalization of values of the markers. These markers may therefore be reliable indicators of therapeutic success. However, these markers underwent different respective changes during the first 2 months. SP-A reached a maximum at the start of the treatment, while SP-D and KL-6 peaked at 5 and 10 days, respectively, after the treatment was initiated. This discrepancy demonstrates that the markers reach the bloodstream by diverse mechanisms and are useful for analyzing pathophysiological alterations in the lung in the early stages of treatment. PMID- 11481833 TI - [A case of ectopic pulmonary calcification appearing as diffuse ground-glass attenuation on HRCT]. AB - A 50-year-old man was admitted to undergo an operation for aortic stenosis and insufficiency. Chest high-resolution CT (HRCT) showed diffuse areas with ground glass attenuation in both lung fields. The shadows were not seen in the periseptal zones. A transbronchial lung biopsy specimen revealed ectopic interstitial calcification. Differential diagnosis upon observation of diffuse areas with ground-glass attenuation on HRCT included hypersensitivity pneumonitis and bronchopneumonia. In the peripheral region of the secondary pulmonary lobules, the microchemical environment of the alveoli tended to become alkalosed in comparison with the central area, the characteristic distribution of the areas of ground-glass attenuation may be explained by the differentiation of the acid base balance between the peripheral and central regions of the secondary pulmonary lobules. PMID- 11481834 TI - Surgery for acute type A aortic dissection using retrograde cerebral perfusion. AB - OBJECTIVE: We reviewed the surgical management of acute type A aortic dissection between 1989 and 1998. METHODS: Subjects were 28 consecutive patients (mean age: 61.8 +/- 10.7 years) with acute type A aortic dissection were studied. The mean duration between aortic dissection onset and surgery was 17.5 +/- 17.0 hours. In surgery, aortic pathology and flow patterns in dissected aortic channels were evaluated using transesophageal and epiaortic echo. Simple, safe combination of profound hypothermic circulatory arrest with retrograde cerebral perfusion and open aortic anastomosis was used for brain protection. Hypothermic circulatory arrest was 46.9 +/- 24.8 minutes. Aortic repair consisted in ascending aortic replacement in 5 patients, with hemiarch repair in 17, and total arch repair in 6. Intimal tears were resected in all but 2 patients. Concomitantly resuspension of the aortic valve was done in 9 and aortic root replacement in 2. RESULTS: No operative (30-day) deaths occurred, although 2 died from unrelated hepatic failure during hospitalization or late-stage pancreatic cancer in the late stage. In cerebral sequellae, 1 patient suffered a stroke and 2 patients developed temporary neurologic dysfunction. CONCLUSION: Our experience demonstrated that the simplified conjunction of hypothermic circulatory arrest with retrograde cerebral perfusion and open aortic anastomosis, associated with real-time assessment by transesophageal and epiaortic echo, is safe and useful during emergency aortic repair for acute type A aortic dissection. PMID- 11481835 TI - Evaluation of an internal thoracic artery as a coronary artery bypass graft by intercostal duplex scanning ultrasonography. AB - OBJECTIVES: Although angiography is often used to determine whether the internal thoracic artery is appropriate as a coronary bypass graft, but use of duplex scanning ultrasonography for this purpose is not yet widespread. METHODS: The internal diameter and flow of the internal thoracic artery were measured using intercostal duplex scanning in 100 patients during April 1995. The ultrasonographic device (sonos 2000, Hewlett Packard) used had a linear probe delivering a frequency of 7.5 MHz. Bilateral internal thoracic arteries and their blood flow were imaged clearly in all subjects. Diameter was compared by angiography and duplex scanning ultrasonography in 20 patients. RESULTS: The average internal diameter of internal thoracic artery was 2.19 +/- 0.46 mm (right) or 2.13 +/- 0.32 mm (left) in men and 2.05 +/- 0.44 mm (right) or 2.09 +/ 0.42 mm (left) in women. The gender difference was statistically significant (p = 0.05). The maximum systolic blood flow velocity through the internal thoracic artery was 0.85 +/- 0.34 m/s (right) or 0.84 +/- 0.36 m/s (left) in men and 0.87 +/- 0.28 m/s (right) or 0.82 +/- 0.28 m/s (left) in women. The average internal thoracic arterial blood flow (F) was 54.6 +/- 29.0 ml/min (right) or 50.9 +/- 28.8 ml/min (left) in men and 56.8 +/- 38.2 ml/min (right) or 58.2 +/- 33.4 ml/min (left) in women. Duplex scanning ultrasonography using an intercostal approach enables easy imaging of bilateral internal thoracic arteries and visualizes entire internal thoracic artery structure by simply changing the probe position. CONCLUSION: Intercostal duplex scanning ultrasonography is thus recommended for reliable evaluation of the internal diameter and blood flow of the internal thoracic artery. PMID- 11481836 TI - Changing predictors of postoperative mortality in acute type A aortic dissection. Is only coronary artery compromise significant? AB - OBJECTIVES: Rapid emergency transport and early diagnosis and surgical treatment for acute type A aortic dissection have improved postoperative survival, which has, however, plateaued at about 80%. End-organ malperfusion is regarded as a strong predictor of postoperative mortality, replacing factors such as cardiac tamponade complications, aortic rupture, and left ventricular dysfunction due to aortic insufficiency. It is thus important to reevaluate risk factors for surgical death to assess current therapeutic strategies. METHODS: We statistically analyzed potential risk factors for perioperative death in 88 patients undergoing surgical repair for type A aortic dissection between January 1990 and December 1999. RESULTS: Univariate analysis showed that cardiopulmonary arrest (adjusted odds ratio: 13.78; p < 0.01) and malperfusion of more than 1 vital organ (adjusted odds ratio 4.97, p < 0.01), especially myocardial ischemia due to coronary artery dissection (adjusted odds ratio 3.21, p < 0.05), significantly increased the likelihood of operative death. Multivariate logistic regression analysis showed only cardiopulmonary arrest (p < 0.01) and concomitant coronary artery bypass grafting necessitated in cases complicated by evolving myocardial infarction (p < 0.05) to be independent predictors of postoperative mortality. CONCLUSION: Preoperative complication from coronary dissection was the most important predictor of early postoperative mortality in this series. In such cases, rapid surgical intervention before myocardial infarction develops is vital to saving lives. PMID- 11481837 TI - Aortic valve repair in dominant aortic regurgitation. AB - OBJECTIVE: We studied immediate and mid-term results after aortic valve repair. METHODS: Immediate and mid-term results were studied in 63 patients undergoing aortic valve repair, calculating survival and reoperation free rates. RESULTS: Subjects were 49 men and 14 women aged 15 to 76 years (mean: 53 +/- 17 years). Mean preoperative aortic regurgitation grading of 1 to 4 was 3.2 +/- 0.7. Mean preoperative New York Heart Association functional class was 1.9 +/- 0.8. Two in hospital deaths occurred. (3.2%) Mean aortic regurgitation grade at discharge was 1.3 +/- 0.8 (p < 0.0001; vs preoperative grade) and functional class was 1.1 +/- 0.2 (p < 0.0001; vs preoperative class),--significantly improved. Overall follow up was 98.4%, and mean follow-up continued 41.4 +/- 22.1 months. Mean functional class at follow-up was 1.2 +/- 0.4 (n = 49), improved from preoperative class (p < 0.0001). Mean aortic regurgitation grading at follow-up was 1.8 +/- 0.8 (n = 41), improved from preoperative grading (p < 0.0001). Five-year survival was 95.1 +/- 2.8%. One-year reoperation freedom was 96.6 +/- 2.4% and 5-year 77.9 +/- 6.9%. CONCLUSIONS: Survival after surgery was good, while reoperation was comparable to other reports but less satisfactory compared to reoperation freedom after aortic valve replacement. Based on reoperative findings, a change in indication was made. We believe technical refinements could improve postoperative results. PMID- 11481838 TI - Port-access cardiac surgery. Experience with 34 cases at Keio University Hospital. AB - OBJECTIVES: We reviewed our experience with port-access cardiac surgery and evaluated the medical effects and benefits of this technique in view of postoperative quality of life and medical expenses incurred during hospitalization. METHODS: From June 1998 to August 2000, port-access cardiac surgery was conducted on 34 patients--22 with atrial septal defect, 6 with mitral regurgitation, 2 with coronary artery disease, 2 with partial endocardial cushion defect, 1 with ventricular septal defect, and 1 with atrial and ventricular septal defects. Two types of endoaortic-balloon catheters were used to execute aortic cross-clamping. Skin incisions were 5 cm long. RESULTS: No hospital or late deaths were observed. Patients with atrial septal defect were discharged on postoperative day 3.7, patients of mitral regurgitation on postoperative day 4.2, and patient of ventricular septal defect on postoperative day 4.0 on the average. None were readmitted. Patients appeared undisturbed by early discharge and were able to resume physical work on day 22 on the average after discharge. CONCLUSION: Patients undergoing port-access cardiac surgery recovered quickly from surgery and resumed work quickly. This technique thus proved satisfactory both physically and mentally to patients and improved their quality of life. Medically and economically this technique proved extremely beneficial. We confirmed it to constitute a viable approach and option for cardiac surgery in selected patients. PMID- 11481839 TI - Concomitant aortic arch and root replacement using a stentless xenograft for acute type A dissection. AB - A 78-year-old woman with an aortic root aneurysm and aortic regurgitation developing acute type A dissection successfully underwent aortic root replacement using a stentless xenograft during core cooling, followed by total aortic arch replacement under selective cerebral perfusion with deep hypothermia. This bioprosthesis can be used in the same way as a free-hand homograft and potentially provides an aggressive, safe option for acute aortic dissection requiring aortic root reconstruction in elderly patients. PMID- 11481840 TI - Ascending aorta dissection associated with bicuspid aortic valve. Considerations 4 years after combined coronary artery bypass grafting and mitral valve replacement. AB - Aortic dissection etiology involve many factors that are difficult to identify clearly. We report a 47-year-old man who underwent a Bentall operation with reattachment of bypass grafts for a dissecting aneurysm (DeBakey type II) 4 years after combined triple coronary artery bypass grafting and mitral valve replacement. This case appeared to be associated with factors leading to dissecting aneurysm although it remains unclear which was more influential congenital bicuspid aortic valve or proximal anastomosis of venous grafts or both. This case suggests the need to consider appropriate timing in surgical intervention for cases of congenital bicuspid aortic valves and the selection of additional aortic valve replacement in initial surgery. PMID- 11481841 TI - Mitral valve replacement through right thoracotomy after coronary arterial bypass grafting with functioning conduits. AB - A 67-year-old man who had undergone coronary artery bypass grafting 3 years previously suffered from severe mitral regurgitation associated with Streptococcal infective endocarditis. He was placed in New York Heart Association functional class III. Preoperative angiography demonstrated good opacification of all 3 conduits implanted in the previous operation. We replaced the mitral valve through an anterolateral right thoracotomy, approaching the mitral valve as an alternative to redoing sternotomy to minimize potential injury to patent grafts. His postoperative course was uneventful. After a 1-month course of antibiotics, the patient was discharged as New York Heart Association class II and at present, 3 months after discharge, is doing well. This approach is an effective alternative to redoing sternotomy for mitral valve operation, especially in patients undergoing a previous coronary arterial bypass grafting via median sternotomy. PMID- 11481842 TI - Tonsillar metastasis from large cell carcinoma of the lung. AB - A 52-year-old female was referred to our department for treatment of a left lung tumor, 80 mm in diameter, arising in the left S1 + 2. The patient's chief complaint was persistent dry cough and spiking fever. Left upper lobectomy with hilar and mediastinal lymph node dissection (ND2a) was performed, and the pathological diagnosis was primary large cell carcinoma of the lung, p-T3N0M0. At one week after being discharged, the patient visited our outpatient clinic complaining of a sore throat. A tumor in the right tonsil was discovered, and excisional biopsy revealed it to be metastasis from the large cell carcinoma of the lung. Right cervical lymph node metastasis was also detected, and the patient was treated by combined chemo-radiotherapy, resulting in a complete remission. PMID- 11481843 TI - Transcatheter coil closure of large patent ductus arteriosus with 0.052-inch Gianturco coils using myocardial biopsy forceps in a 70-year-old woman. AB - Small patent ductus arteriosus is generally closed in children using a transcatheter coil. This is done less often in older patients or those with large patent ductus arteriosus. We report successful antegrade transcatheter coil closure of patent ductus arteriosus in a 70-year-old woman. Into the patent ductus arteriosus, using flexible myocardial biopsy forceps, we placed two large 0.052-inch Gianturco coils, which were easily used as multipurpose vascular occlusion coils. The forceps and the coils were readily available and provided complete occlusion. Other delivery devices cannot deliver such large coils. Transcatheter coil closure thus appears to be safe and effective for closing large patent ductus arteriosus in the elderly. PMID- 11481844 TI - Mediastinal neurilemmoma complicated with spinal subarachnoid hemorrhage. AB - A 28-year-old woman suffered severe back pain and headache during exercising on three occasions during the prior two-month period. On admission, the physical examination revealed symptoms of meningeal irritation, nuchal rigidity, severe headache, continuous nausea, and vomiting. Cerebral computed tomography of the intracranial subarachnoidal space revealed no subarachnoid hemorrhage. Her cerebrospinal fluid was bloody. Spinal magnetic resonance imaging identified a posterior mediastinal tumor adherent to the left side of the 5th thoracic vertebra and an abnormally expanded blood vessel near the mediastinal tumor. In addition, a high signal intensity lesion appeared to be present on the surface of the spinal cord. A mediastinal neoplasm was removed through standard thoracotomy. During surgery, marked enlargement was noted in some veins (hemiazygos and 5th intercostal veins) which apparently had been constricted by the mediastinal tumor. Surgical and radiological findings suggested a relationship between the constricted venous return due to the tumor and the patient's spinal subarachnoid hemorrhage. PMID- 11481845 TI - Jaundice after surgery for an aortic arch aneurysm. AB - A 57-year-old patient who developed hyperbilirubinemia after surgery for an aortic arch aneurysm subsequently suffered pseudomonas sepsis. Low-volume biliary drainage from the common bile duct was colorless. A disturbance in the liver's excretory system caused jaundice. Sepsis and jaundice were resolved when hepatic excretory function recovered. PMID- 11481846 TI - Indication and perioperative management for cardiac surgery in patients with liver cirrhosis. Our experience with 3 patients. AB - Three patients with noncardiac Child A cirrhosis underwent cardiac surgery. All survived surgery, but 2 died during follow-up periods. A 61-year-old woman who underwent successful double valve replacement died of diabetic coma and severe acidosis due to intestinal necrosis 18 months later. A 57-year-old woman who underwent successful mitral valve replacement died of liver failure induced by heart failure 9 years later. A 45-year-old man who underwent coronary artery bypass grafting is doing well 18 months after discharge. Proper perioperative management, including high-flow cardiopulmonary bypass, pharmacological and mechanical circulatory support, and mechanical respiratory support prevented further, potentially lethal, hepatic dysfunction, leading to good early surgical results. We concluded that patients with Child A cirrhosis could tolerate cardiac surgery. Subsequent surgical results, however, were unsatisfactory, and more careful follow-up is necessary to obtain better late results. PMID- 11481847 TI - Late aortic dissection in a patient with a left ventricular assist device. AB - The limited availability of donor hearts means that an increasing number of transplantation candidates are temporarily or permanently supported by mechanical circulatory assist devices. We report a patient undergoing implantation of a Novacor left ventricular assist device who suffered fatal aortic dissection on postoperative day 11 after satisfactory recovery from multiple organ failure. The dissection of the aorta initially presented as an embolic peripheral ischemia. Early complete echocardiography is thus warranted to rule out dissection. PMID- 11481848 TI - Single lung transplantation from a brain-dead donor for a patient with idiopathic pulmonary fibrosis. A breakthrough after new legislation in Japan. AB - Two single lung transplants from a single cadaveric donor were successfully conducted at 2 institutions on March 29, 2000, the first such procedure in Japan under newly introduced legislation. Our patient was a 48-year-old woman with idiopathic pulmonary fibrosis who underwent left single-lung transplantation under cardiopulmonary support. The donor lung was preserved in 4 degrees C modified Euro-Collins solution. Total ischemic time was 5 hours and 37 minutes. The postoperative course was uneventful. The patient was discharged on postoperative day 62 with satisfactory respiratory function. PMID- 11481849 TI - Extended operation for non-small-cell lung cancer invading into the liver. PMID- 11481850 TI - [Changes of muscle excitability of the hand contralateral to a task performing one: a transcranial magnetic stimulation study]. AB - It used to be considered that unilateral movements of distal limb parts are associated only with contralateral motor cortical activity. Recent neuroimaging studies, however, suggest that the motor cortex ipsilateral to a task-performing hand is also activated, and that motor patterns in one hand affect the degree of the activity of the ipsilateral motor cortex. If so, muscles of the hand contralateral to a task-performing one may change those excitability depending on types of tasks. We studied eight subjects who performed three different finger tasks by one hand: (a) pinch, (b) sequential finger opposition, and (c) tactile discrimination. Transcranial magnetic stimulation was delivered by a figure eight coil over the hemisphere ipsilateral to a task-performing hand. Motor evoked potentials and background electromyographic activities were recorded from the opponens pollicis muscle contralateral to the stimulated hemisphere. On average, the motor evoked potentials were larger during tactile discrimination task than those at rest in either hand (p < 0.01). Background electromyographic activities in the left hand increased significantly during right hand tactile discrimination task (p < 0.01), whilst those in the right hand did not change during the left hand performance (p > 0.05). These findings suggest the followings: (1) the hand muscle contralateral to a task performing one changes its excitability depending on types of tasks; and (2) increment of excitability of the left hand muscle associated with right hand tactile discrimination is greater than that of the right hand one in association with the same task by the left hand, thus supporting the idea that there is a functional asymmetry between the right and left motor cortex in respect of motor performance. PMID- 11481851 TI - [Intravascular malignant lymphomatosis: an autopsy case with generalized telangiectasia and various neurological manifestations]. AB - We report a 73-year-old woman with intravascular malignant lymphomatosis (IML) who showed generalized telangiectasia as well as various neurological symptoms. In July 1998, she developed fever, dizziness, and confusion followed by left hemiparesis, and was admitted to our hospital on August 11, 1998. Laboratory tests indicated a normochromic normocytic anemia, thrombocytopenia, elevated serum lactic dehydrogenase (LDH), C-reactive protein (CRP), and cerebrospinal fluid protein. Magnetic resonance imaging (MRI) of the brain revealed an infarct like lesion in the left frontal lobe and multiple white matter lesions. After admission, her neurological status deteriorated and lapsed into coma and quadriplegia. At the end of September 1998, generalized telangiectasia appeared, and she was diagnosed as IML on skin biopsy. Although combination chemotherapy failed to improve her neurological symptoms, telangiectasia disappeared in a few days, and the infarct-like lesion on MRI decreased in size. Serum LDH, CRP, and thrombocyte counts were normalized. Autopsy findings revealed perivascular clustering of B cell type lymphoma cells in the left frontal lobe where abnormal signal intensity was found on MRI, as well as the spleen and the bone marrow. This case emphasizes the importance of early diagnosis and treatment in IML. PMID- 11481852 TI - [Repeated hyperglycemic hemichorea in a patient with venous angioma in the putamen]. AB - We report a case of an 81-year-old diabetic woman who had three episodes of choreic involuntary movement in the left extremities. Brain CT revealed faintly increased density in the right putamen. On MRI performed 28 days after onset of the left hemichorea, the right putamen showed increased signal intensity on T1 weighted images and the so-called "medusa-like appearance" of medullary venous drainage into the thalamostriate vein on enhanced T1-weighted images. These findings indicated petechial hemorrhage of the right putamen from a venous angioma. We hypothesize that this hemorrhagic change due to a venous malformation in the basal ganglia may induce involuntary movements in diabetic patients during severe hyperglycemia. PMID- 11481853 TI - [A case of mononeuritis multiplex associated with type I (monoclonal) IgG kappa cryogloblinemia]. AB - We here reported a fifty-three year-old woman with mononeuritis multiplex, associated with type I cryoglobulinema induced IgG kappa MGUS (monoclonal gammopathy of undetermined significance). She first experienced numbness in the extremities, that showed stepwise deterioration for several weeks. The nerve conduction study revealed axonal neuropathy, and laboratory examination detected type I cryoglobulinema. The sural nerve biopsy demonstrated with severe axonal degeneration with focal accentuation, lacking any inflammatory response. Single cryofiltration definitely stopped the progression of the symptoms. The prompt clinical response to the removal of cryoglobulin, without the administration of anti-inflammatory drugs, confirmed the direct etiological correlation between type-1 cryoglobulin and the neuropathy, and may favor ischemic, rather than inflammatory nature of this rare type of neuropathy. PMID- 11481854 TI - [A patient with Crow-Fukase syndrome associated with pulmonary plasmacytoma]. AB - We here reported a 54-year-old female patient with Crow-Fukase syndrome associated with pulmonary plasmacytoma. She was found to have scattered tumor in 1990. Although the tumor had slowly grown for the last 10 years, she showed no clinical symptoms. Numbness and weakness of lower extremities began in June 1999, and she was referred to Kyoto University Hospital on Oct. 21 1999 for evaluation of progressive symptoms. She had skin pigmentation, edema of the lower extremities, lymphadenopathy, muscle weakness and sensory disturbance in a glove and-stocking distribution. Serological examination showed monoclonal IgG-lambda gammopathy. Serum vascular endothelial growth factor (VEGF) was markedly elevated. Microscopic studies on biopsied sural nerve demonstrated mild decrease of myelinated fibers. Immunohistochemically, the pulmonary tumor was defined as an IgG (lambda type) plasmacytoma. After treatment with melphalan-prednisolone therapy, the neurological symptoms improved along with decrease of serum VEGF levels as well as the size of pulmonary plasmacytoma. This is the first report of a patient with Crow-Fukase syndrome associated with pulmonary plasmacytoma. This case suggests that growth of pulmonary plasmacytoma might have played an important role in the overproduction of VEGF and thus development of Crow-Fukase syndrome. PMID- 11481855 TI - [Paradoxical embolism with Chiari network, subsequently being accompanied by probable incomplete infarction: a case report]. AB - We presented a patient of paradoxical embolism with Chiari network, subsequently being accompanied by probable incomplete infarction. This 21-year-old man suffered from consciousness disorder, aphasia and right hemiparesis, and hospitalized in November 6, 1999. Magnetic resonance imaging showed mixed intensity on T1 and T2-weighted images in part of the areas of the left anterior and middle cerebral arteries. Cerebral angiography revealed the early venous fillings and the capillary blushs. These findings implicated stroke in young adult. Still more transcranial color-flow imaging showed high intensity transient signals with "Chirp" sounds on the left middle cerebral artery. Transesophageal echocardiography detected Chiari network. Chiari network was thought the course of cerebral infarction. Over again 123I-IMP single-photon emission CT findings revealed the marked reduction of his cerebral blood flow comprehensively in the left hemispherium. It was suggested that the recanalization after the paradoxical cerebral embolism had caused incomplete infarction. PMID- 11481856 TI - [Anti-GD1b IgG antibody-related Guillain-Barre syndrome initially mimicking brainstem infarction]. AB - We described a 58-year-old woman with Guillain-Barre syndrome, who initially showed rapid progression of brainstem infarction-like signs. She developed superficial sensory disturbance on the left side, dysarthria, and left predominant limb weakness within a few hours. She showed bilateral extensor plantar responses and head CT scan detected no abnormality. It was difficult to be distinguished from brainstem infarction until symmetrical limb weakness and generalized areflexia appeared. Serum anti-GD1b IgG antibody with cross reactivity with GM1b was detected. Cerebrospinal fluid examination revealed albuminocytologic dissociation on day 5. After 5 sessions of immunoadsorption therapy, her symptoms gradually lessened. Anti-GD1b antibody has been detected in patients with sensory ataxic neuropathy. Our patient, however, was characterized with early involvement of brainstem with ataxia of cerebellar type. Our case suggests that anti-GD1b antibody-associated neuropathy has a broad spectrum of clinical features, which are related to cross-reactivity of this antibody. PMID- 11481857 TI - [An adult case of primary antiphospholipid syndrome presenting recurrent anterior spinal artery syndrome]. AB - A 50-year-old man suffered from sudden onset of paraplegia, sensory impairment of the lower extremities and difficulty urinating. These symptoms spontaneously resolved, and he was able to walk again within 10 days. After 2 months' observation, he experienced a relapse with identical symptoms. Neurological examination revealed paraplegia, hypalgesia and hypesthesia in the lower extremities and difficulty urinating. Position and vibration senses were preserved in the extremities. Knee jerk was mildly hyperreflexic on the left, but pathological reflexes were not present. He had such risk factors for arteriosclerosis as diabetes mellitus, hypercholesterolemia and smoking. Although antinuclear antibodies were negative, he repeatedly showed high anticardiolipin antibody titers. MRI showed a T2-hyperintense lesion of the anterior portion of the spinal cord at Th9-10 levels. Neither arteriovenous malformation nor fistula was found on MR angiography. Abdominal CT revealed no abnormality in the descending aorta. Cervical ultrasonogram showed arterioslerotic lesions on the right common carotid artery. This is the first report of recurrent spinal cord infarction in an adult patient associated with primary antiphospholipid syndrome. Recurrence of the spinal cord infarction in this case may have been facilitated by the presence of risk factors for arteriosclerosis. PMID- 11481858 TI - [An autopsy case of Ehlers-Danlos syndrome showing various types of cerebrovascular disease]. AB - We report a 73-year-old woman with Ehlers-Danlos syndrome (EDS) and hypertension who had developed various types of cerebrovascular disease. She had suffered from cerebral hemorrhage of the left putamen at the age of 58, of the left parietal lobe at 64 and cerebral infarction of right internal capsule at 71. EDS type II or III was suggested by two times of skin biopsies. A brain CT at the age of 73 revealed a comparatively large cerebral aneurysm in the territory of the anterior cerebral artery. The patient was treated conservatively, but died due to rupture of the aneurysm. The wall of the aneurysm was made up thin collagen fibers without elastic fibers. There were other multiple small aneurysms in the cerebral arteries, but none in other organs. Deposition of acid mucopolysaccharides was noted in the media of the abdominal aorta. Finally, the present case was thought most likely to be of EDS type IV. It was suggested that one of the causes of the cerebral hemorrhage at the ages of 58 and 64 and the infarction at 71 was related to hypertension, since brain MR angiography at 71 showed no clear aneurysms. In cases of EDS, one should consider the possible formation or rupture of cerebral aneurysm even though the course is favorable. PMID- 11481859 TI - [Report of a patient with CADASIL having a novel missense mutation of the Notch 3 gene--association with alopecia and lumbar herniated disk]. AB - We report a 52-year-old man with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) presenting dementia, alopecia and lumbar herniated disk. He had an episode of stroke and migraine-like headache lasting for 5 minutes. A lot of members had cerebral infarction in this family. Brain magnetic resonance imaging demonstrated, on T2-weighted images, numerous hyperintense lesions suggestive of small infarcts in the basal ganglia and diffuse hyperintense lesions in the cerebral white matter. The clinical symptoms, the family history and the MRI findings suggested the diagnosis of CADASIL. However, the patient also showed alopecia and lumbar herniated disk, both are characteristic features of cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). The DNA analysis of the Notch 3 gene identified a novel missense mutation Cys174Phe in this patient. Our case report indicated the importance of the DNA analysis for the diagnosis of CADASIL. PMID- 11481860 TI - [Severe chronic pain with allodynia in Parkinson's disease: a case report]. AB - We report a 61-year-old man with Parkinson's disease, who had a 3-year history of severe chronic pain with allodynia in the lower extremities prior to motor symptoms. He always had tingling pain around the ankles, and tactile sensation induced severe burning pain expanding to the toes and thighs, so his pain was considered to be allodynia. Pain and motor symptoms were ameliorated by L-dopa therapy and exacerbated by withdrawal of L-dopa. Pain is known to occur in Parkinson's disease, but severe pain rarely occurs. To our knowledge, allodynia, which is usually recognized in causalgia or reflex sympathetic dystrophy, has never been reported in Parkinson's disease. Patients with Parkinson's disease may complain severe causalgia-like pain as an initial symptom. PMID- 11481861 TI - [A case of progressive supranuclear palsy improved with tandospirone citrate]. AB - We reported a 74-year-old male case of progressive supranuclear palsy (PSP) who responded to tandospirone citrate, a serotonin receptor (5-HT1A) agonist. The patient manifested postural instability and gait disturbance at 71 years. Additionally, he showed vertical gaze paresis, regidity of the neck, extremities and trunk, bradykinesia and mild cognitive impairment. A brain MRI revealed moderate atrophy of bilateral frontal/temporal lobes and of midbrain tegmentum one year after the onset. The patient had been diagnosed as PSP and treated with L-DOPA. However, L-DOPA therapy showed only transient response for a few months. His symptoms deteriorated gradually, and he became unable to sit, stand up or walk by himself. Tandospirone citrate was additionally administered at 30 mg/day. Rigidity and bradykinesia were remarkably improved in two weeks after the start of tandospirone treatment. He became able to stand up and walk a short distance with supports in four weeks. Cognitive disturbance was also slightly improved. Tandospirone citrate was effective on our case of PSP, especially on rigidity. Our findings suggest that combination of levodopa and tandospirone citrate is a useful therapy for PSP. PMID- 11481862 TI - [MRI finding of CNS cryptococcosis in an HIV-positive patient]. AB - We report an HIV-positive patient with CNS cryptococcosis, diagnosis of which was based on detection of Cryptococcus neoformans by Indian ink staining and culture of CSF. MRI displayed dilated Virchow-Robin space in bilateral basal ganglia which were hypointense on T1-weighted images, hyperintense on T2-weighted images, and enhanced by gadolinium administration. In addition cryptococcoma in the cerebellum was observed by MRI. This finding may suggest a progression from cryptococcal meningitis to intraparenchymal invasion, accompanied with breakdown of the blood brain barrier. PMID- 11481863 TI - [Adult leigh syndrome with mitochondrial DNA mutation at 8993--mutation ratio analysis on formalin-fixed autopsy tissues]. PMID- 11481865 TI - [Telomerase, cell immortality and cancer]. AB - Telomerase is an enzyme that replaces repetitive (TTAGGG)n sequences on the ends of chromosomes that would otherwise be lost during successive cell divisions. Telomerase activity is closely linked to attainment of cellular immortality, a step in carcinogenesis, while lack of such activity contributes to cellular senescence. Telomerase is activated in more than 85% of malignant tumors. However, with the exception of some self-renewing tissues with high regenerative potential, telomerase activity is usually repressed in normal somatic tissues. Based on these reports, we investigated telomerase activity in gastric mucosal tissues. Telomerase activity is highest in cancer, followed by intestinal metaplasia, chronic gastritis, and normal mucosa. In patients with intestinal type gastric cancer, telomerase activity was higher in those with intestinal metaplasia and H. pylori infection than in patients without infection. Our results suggest that H. pylori infection may influence telomerase activity in cancer and noncancerous tissue. Genes encoding three major components of human telomerase have been recently cloned. They included those for human telomerase RNA component (hTR), human telomerase reverse transcriptase (hTERT), and telomerase-associated protein 1 (TEP1). More recently, two human telomeric repeat binding factors (TRFs) have also been cloned: TRF1, considered to inhibit the action of telomerase at the telomeric region, and TRF2, believed to prevent fusion of chromosome ends and, in vitro, to remodel linear telomeric DNA into large duplex loops. However, the details of mechanisms regulating telomerase activity are still poorly understood, and specific components or binding proteins that might represent suitable targets for cancer gene therapy have not yet been identified. Therefore, we established quantitative assays using a TaqMan RT-PCR for mRNAs encoding the telomerase components hTR, hTERT, and TEP1, as well as for those encoding TRF1 and TRF2. By using our quantitative assays, we found the following results: 1) Expression of TRF1 and TRF2 mRNA was greater in the normal cells than in human malignant hematopoietic cell lines or in patients with acute leukemia, 2) hTERT mRNA expression showed changes paralleling telomerase activity and became undetectable with HL60 cell differentiation, 3) initially low expression of TRF1 and TRF2 mRNA increased during differentiation. Our results suggest that not only hTERT but also TRF1 and 2 are important regulators of telomerase activity. PMID- 11481866 TI - [A behavioral and neurochemical study on the mechanism of the anxiolytic effect of monoamine oxidase inhibitors]. AB - The author examined the acute anxiolytic effects of monoamine oxidase inhibitors on freezing behavior, a putative index of anxiety, induced by conditioned fear stress. The selective serotonin1A receptor agonist inhibited freezing dose dependently. The irreversible, non-selective monoamine oxidase inhibitors tranylcypromine (3 and 15 mg/kg) and phenelzine (30 and 80 mg/kg) reduced freezing significantly. Clorgyline (10 mg/kg, irreversible selective monoamine oxidase A inhibitor), Ro 41-1049 (30 mg/kg, reversible selective monoamine oxidase A inhibitor), selegiline (3 mg/kg, irreversible selective monoamine oxidase B inhibitor) and lazabemide (10 mg/kg, reversible selective monoamine oxidase B inhibitor) had no effect on freezing behavior. However, combined administration of clorgyline (10 mg/kg) and selegiline (3 mg/kg) reduced freezing significantly, as well as combined administration of clorgyline (10 mg/kg) and lazabemide (10 mg/kg), Ro 41-1049 (30 mg/kg) and selegiline (3 mg/kg), or Ro 41 1049 (30 mg/kg) and lazabemide (10 mg/kg). These effects of monoamine oxidase inhibitors on freezing were not due to non-specific motor effects. These results suggest that acute inhibition of both monoamine oxidase A and B reduces anxiety or fear, while inhibition of monoamine oxidase A or B alone fails to reduce anxiety or fear. In vivo microdialysis studies showed that the irreversible monoamine oxidase A inhibitor clorgyline and the irreversible monoamine oxidase B inhibitor selegiline induced a mild increase and no increase in extracellular serotonin, respectively. Interestingly, the combined treatment with clorgyline and selegiline resulted in much larger increases in extracellular serotonin in the medial prefrontal cortex than did either monoamine oxidase inhibitor alone. Our previous studies have indicated that facilitation of 5-HT neurotransmission decreases conditioned freezing, i.e., anxiety or fear. The results of these in vivo microdialysis studies may account for the results of this study that the simultaneous blockade of both monoamine oxidase A and B reduced conditioned freezing, whereas blockade of either monoamine oxidase alone failed. PMID- 11481867 TI - [Relationship between the expression of truncated gelsolin and invasiveness in malignant melanoma]. PMID- 11481868 TI - [A study on the contact between tectorial membrane and inner hair cell stereocilia in the cochlea]. PMID- 11481869 TI - [Analysis of a human synovial sarcoma-associated chimeric oncogene product]. PMID- 11481870 TI - [p53 gene mutations in in situ pancreatic adenocarcinomas]. PMID- 11481871 TI - [Expression of tyrosine kinase syk in pancreatic cancers]. PMID- 11481873 TI - [Hokkaido University Neuroscience Conference 2000]. PMID- 11481872 TI - [The WHO's Leprosy Elimination Programme and the Japanese staff in WHO--part of public health training programme at Hokkaido University School of Medicine]. PMID- 11481874 TI - Biocompatibility of root canal filling materials. AB - Results of in vitro and in vivo studies clearly indicate that some endodontic sealers may cause local and systemic adverse effects. Though occasionally contradictory data has been reported from various authors, it may be concluded that zinc-oxide-eugenol sealers possess a marked cytotoxic and tissue-irritating potency. Most Ca(OH)2-based materials, however, were biocompatible. Genotoxic effects have been observed with sealers releasing paraformaldehyde or containing mutagenic substances, such as bisphenol-A-diglycidyl-ether or its derivatives. It cannot be excluded that these materials may pose a systemic risk because formaldehyde is rapidly distributed systemically following its application into the pulp cavity. Furthermore an increasing number of cases with an aspergillosis of the maxillary sinus have been observed which were mainly caused by zinc releasing endodontic sealers. Overall, it is recommended that for endodontic practice, sealers that have been found to be biocompatible in a "mixed bag" of various in vitro and in vivo tests, be selected. From this point of view, ZnOE sealers should no longer be used for root canal fillings. This recommendation applies also to sealers containing paraformaldehyde or generating this substance during their setting reaction. More experimental and clinical studies are necessary to elucidate whether new materials, such as mineral trioxide aggregate (MTA) or calcium phosphate cement, will be biocompatible alternatives in the future. PMID- 11481875 TI - Anatomical variation of the sphenomandibular ligament. AB - The sphenomandibular ligament, which is derived from the sheath of Meckel's cartilage, is a fibrous structure that passes between the spine of the sphenoid bone and the lingula of the mandible. Although anatomical texts provide basic descriptions of this structure, there are few published reports of the extent of its variability or its possible clinical implications. The aim of this study was to provide a detailed description of the nature and extent of anatomical variability in the sphenomandibular ligament of seven human cadavers. Dissections of sagittally sectioned heads were performed using a medial approach that involved displacement of the tongue and mucosal tissues in the oral cavity and oropharynx, then reflection of the medial pterygoid muscle. The ligaments ranged in shape from thin bands that descended for a short distance from the spine of the sphenoid to broad bi-concave ligaments with prominent insertions. The mylohyoid nerve was seen to pass behind the ligament in all specimens, emerging from the postero-inferior border of the mandibular attachment before running into the mylohyoid groove on the medial surface of the ramus. Lying in the pterygomandibular space, the ligament was surrounded by fascia, both structures presenting potential barriers to the diffusion of local anaesthetic solution if injected medially. The appearance of the lingula also varied, and did not seem to reflect the size of attachment of the ligament, suggesting an alternative explanation of lingula morphology, perhaps related to a continuation of the mylohyoid ridge and anterior border of the mylohyoid groove. PMID- 11481876 TI - A unilocular odontogenic keratocyst associated with the periradicular area of an upper right maxillary canine--a case report. PMID- 11481877 TI - Conservative management of an intruded immature maxillary permanent central incisor with healing complication of pulp bone. AB - Traumatic intrusion injury of permanent teeth is serious with multiple complications possible associated with the pulp, periodontal ligament, alveolar bone and Hertwig's epithelial root sheath. The optimal treatment for the management of an intrusion injury has not yet been determined. A case is presented involving the conservative management of an immature maxillary permanent central incisor intrusively luxated by allowing for re-eruption and orthodontic extrusion two weeks later. After a follow-up period of ten months, the intruded tooth continued to show a mobility of grade one, without metallic percussion tone or infra-occlusion, which confirmed periodontal ligament healing. Although the intruded tooth failed to respond to dry ice testing, no other signs of pulp necrosis were evident and the colour of the intruded tooth was within normal limits throughout the follow-up period. However, complications of healing of Hertwig's epithelial root sheath occurred, causing in-growth of bone and periodontal ligament into the root canal. PMID- 11481878 TI - Apical sealing ability of five endodontic sealers. AB - Because apical leakage may be a cause of failure of endodontic treatment, the purpose of this study was to compare the apical seal provided by five endodontic sealers. After cleaning and shaping procedures, the root canals of 75 human cuspids were obturated by lateral condensation using gutta-percha and the following sealers: Kerr Pulp Canal Sealer EWT, Grossman's sealer, ThermaSeal, Sealer 26, and AH Plus. These root-filled teeth were then maintained at 37 degrees C and 100% humidity for two days. After immersion in a solution containing black India ink and fetal bovine serum for five days, the teeth were demineralised and rendered transparent. The extent of dye penetration was examined under a stereomicroscope. The results showed that Grossman's sealer exhibited significantly more leakage than the other sealers, except ThermaSeal. There was no significant difference between ThermaSeal and AH Plus. No significant differences were observed for Kerr Pulp Canal Sealer EWT when compared with either ThermaSeal or AH Plus. There was significantly less leakage for Sealer 26 when compared with the other sealers tested, except AH Plus. PMID- 11481879 TI - Maxillary lateral incisor with two roots. AB - Variations in root canal morphology can lead to complications in endodontic treatment. A case of a maxillary lateral incisor with two roots is reported. PMID- 11481881 TI - Virtual Web wave of the future: integration of healthcare systems on the Internet. PMID- 11481882 TI - Use of information technology by a client-centered provider. A case study of Harvard Vanguard Medical Associates. PMID- 11481883 TI - Patients' rights legislation back in the spotlight. PMID- 11481885 TI - Preparing for a career in medical management. 8 steps to help you evaluate your career path. AB - Making a career switch from clinical practice to medical management requires preparation and planning. There are numerous steps you can take to smooth out the transition. Most importantly, you must find ways to gain valuable leadership and management experiences before you can expect to land a good medical management position. Discover ways to cultivate that experience through staff posts, part time positions or volunteer work. PMID- 11481886 TI - Physician executives breaking out of middle management. AB - Physician executives are slowly moving into top management jobs, holding the same stature on senior management teams as chief financial officers and executive vice presidents. Learn how the role of the physician executive is evolving and how opportunities are expanding for even more physicians to assume the top-level posts. PMID- 11481887 TI - Medical director musical chairs. A reaffirmation of my career in medical management. AB - Losing your job can be a traumatic event. But you can minimize the shock by taking some action steps to find another job quickly--in many cases, even a better job. Consider some advice from a seasoned physician executive who has successfully negotiated the corporate maze found in many health care organizations today. PMID- 11481888 TI - Unconventional style reaps rewards for Ruckdeschel. Physician executive's career carries cancer center to the forefront. Interview by Patty Ryan. AB - He didn't like math. Loved biology. So he ditched his plans to become an engineer and ended up pursuing a career in medicine and hospital administration. He led a sleepy cancer center to new heights of cutting-edge research and progressive types of treatment. And he did it all on his own terms--a mix of practicality and instinct that serves up some interesting perspectives for fellow physician executives to consider. Meet John Ruckdeschel, MD. PMID- 11481889 TI - Working with recruiters. Physicians sound off in CyberForums about their experiences with recruiters. AB - Searching for your next job is never easy. You generally need all the help you can get. Often, that means working with physician recruiters who have the connections to help you land a great position. But dealing with recruiters can be both rewarding and frustrating. In two ACPE CyberForums facilitated by Barry Herman--one in 1999 and another just a few months ago--participants talked about the role of recruiters, offering advice and tips that can help physician executives carry out a successful job search. PMID- 11481890 TI - Negotiating compensation packages. Bargaining to get the best offer. AB - You worked hard to search for a job--sending out resumes, networking with your peers and contacting recruiters. But when the offer finally comes, your work isn't done. Now it's time to negotiate the best salary and employment contract. Take a look at today's standard compensation packages, including bonuses, stock options, relocation expenses and other features. What's negotiable and what's not? PMID- 11481891 TI - Controlled burn! Physician executives must be ready to handle job burnout, career stress. AB - Job burnout for workers in any career can be frightening. It's truly debilitating to think that your job and career are worthless, that your future isn't bright. There are, however, some steps you can take to recognize, avoid or overcome the stress and anxiety that leads to job burnout. PMID- 11481892 TI - Physician executives report high job satisfaction. Summary of findings from a survey of senior physician executives. AB - The role of the senior physician executive is well established in American hospitals and health systems. There is little research, however, on overall physician executive job satisfaction, their perceptions of their organizational role and job performance, or their views of the medical staffs with which they work. A recent survey of physician executives examined these and other areas. It found physician executives to be quite satisfied with their jobs. What follows is a summary of the findings. An article based on the survey will be featured in a future issue of The Physician Executive. PMID- 11481893 TI - The role of the health care leader. Past, present, future. AB - Strong, effective communication may be the single most important key to success for any type of executive. Leaders of health care organizations must be able to promote ideas that others will implement, help staff see the value of their work, and have the vision to overcome limitations that stand in the way of progress. PMID- 11481894 TI - The metabolism of change. AB - A flood of new technologies is engulfing health care. Adopting them can be magical, or horrifically problematic. Yet few institutions have any real, routine process for thinking about them. There are four essential actions an organization must take to explore this fascinating, confusing world of high technology. PMID- 11481895 TI - Timing and the transitory nature of biotech startups. PMID- 11481896 TI - What do you owe your team? Survival tips for people who dread teamwork. AB - Team building and teamwork are well entrenched in the American workplace. If you're independent, shy, or short on time, making decisions with a team may be one of the toughest situations you face. Learn how to get along with the rest of the group even if you don't buy in to the team concept. Teamwork may be one of your most useful career skills. PMID- 11481897 TI - Federal budget helps drive health policy. A close-up look at the budget process and how it affects health care funding. AB - Crafting the budget is an annual process that Congress and the administration use to define the national policy agenda. It is a massive undertaking to decide how more than two trillion dollars is spent by the federal government. Over 20 percent of the budget is used for health care. The budget is a political document that defines the priorities of the federal government and directly influences overall health policy. Its construction deserves to be followed and influenced by physicians to ensure the health of the public. PMID- 11481898 TI - Sage advice. Experienced physician executives share the secrets of their success. Interview by David O. Weber. AB - Seven successful physician executives, now retired from distinguished careers, look back over their experiences and offer advice for those who follow. Be a good doctor first, and never let business interests undermine the ethics of the health care profession. Prepare for management responsibilities with business training. Seek out mentors and develop a network of knowledgeable colleagues and advisors. Exert leadership rather than authority. Expect disappointments and arm yourself for them. PMID- 11481899 TI - New cardiac project is first to tackle clinical and operational issues. AB - Any hospital with a cardiac program has been involved in a benchmarking project that focuses on clinical outcomes. But to date, no one has looked at both clinical and operational issues on any scale. Now the University Hospital Consortium (UHC) based in Oak Brook, IL, is about to do just that. PMID- 11481900 TI - Rapid-cycle feedback can work for multiple needs. AB - Why wait for data when you can have them now? For a dozen years, management at Delnor Community Hospital in Geneva, IL, saw patient satisfaction scores fluctuate between the 50th and 75th percentile. And for a dozen years, management felt that while that was 'pretty good, we wished we could do better,' says Michael Kittoe, MBA, the chief financial officer of the 118-bed facility. About two years ago, the hospital started a campaign to make customer service a real competitive differentiation for the hospital. PMID- 11481901 TI - Survey results spur spiritual care program. AB - Simple ideas win national recognition. There have been plenty of studies in the last few years that indicate patients do better if their spiritual needs are being met. That is the message that Baxter Regional Medical Center in Mountain Home, AR, recently took to heart. PMID- 11481902 TI - Benchmarking: beating the information explosion. AB - What could possibly be more intriguing for health care professionals than access to comparative data, of all types, for use at any time? The only fantasy more engaging would be having the information and actually being able to use it constructively. PMID- 11481903 TI - Has the nursing shortage decreased health care quality? PMID- 11481904 TI - Study targets errors in ambulatory setting. PMID- 11481905 TI - [Human papillomavirus and cervical cancer: genetic background of the neoplastic process]. AB - In a 2-year period, 136 HPV positive cytological samples of the cervix uteri were analyzed at the Department of Molecular Pathology, National Institute of Oncology, Hungary. Comparison with the international data obtained from the literature revealed that the Hungarian epidemiological data bore closest resemblance to the European ones except some differences. The HPV18 is rather seldom encountered in this country. Similarly low occurrence was noted only in Japan. However, the 14.1% occurrence rate of HPV58 in Hungary is by far higher than that in any other country in this analysis except Japan where this virus is of similarly high frequency. In Hungary, the incidence of HPV59 is relatively high just like in Central and South America. HPV33 and HPV66 infections occur in a significantly higher number with Hungary than in any of the countries studied. In our study The European type variant of HPV16 (E-V-350G) occurred in 2/10 CIN II-III cases. The authors also compared the various clinico-pathological grouping of HPV types published, and identified several inconsistencies. Viruses considered to have high risk occurred in intact epithelium, CIN I-II-III and carcinoma alike. The general tendency was, however, that certain viruses correlated with specific clinico-pathological entities. At present there is no reason to include the PCR-based HPV typing in the mass screening of cervical cancers. HPV typing and physical state of the virus can reasonable be determined if the cervical cytology is suspect for HPV infection or even control examination after "loop" conisation. Negative cytology completed with negative HPV-DNA test means the lack of cancer risk even in the case of a previously removed CIN or carcinoma. However, a positive HPV test detected after conisation associated with negative cytology finding indicates a risk of 70% of the development of CIN within 2 years. PMID- 11481906 TI - [Homocysteine--a risk factor for atherosclerosis]. AB - Elevated plasma homocysteine is a new risk factor for atherosclerotic vascular disease resulting in progressive atherogenesis in the arteries of the limbs, the coronary arteries and the cerebrovascular system. Hyperhomocysteinemia may be induced by failure or decreased enzyme activity of the cystathionine-beta synthase and methylenetetrahydrofolate reductase due to genetic mutation or deficiency of folic acid, vitamin B12 and vitamin B6. Oxidation of homocysteine to homocystine is accompanied with production of hydrogen peroxide inducing damage of endothelium through oxidative stress. The injury of the endothelium by homocysteine can be shown by measuring flow-induced vasodilation in men. The abnormalities of coagulation found in hyperhomocysteinemia is related to the impairment of the function of endothelial cells and inhibition of the thrombomodulin-protein C and glycosaminoglycan-antithrombin-III anticoagulant system. Homocysteine decreases the level of glutathione peroxidase in the endothelial cells, and inhibits its activation leading to the impairment of oxidative defensive mechanism, and to the free radical-induced NO-inactivation. In decreasing of plasma homocysteine level and preventing its influence on endothelium, moreover in improving of endothelial function the folic acid has cardinal importance, however the vitamin B12 and vitamin B6 also play role in the maintenance of normal homocysteine level of blood. PMID- 11481907 TI - [Vascular diseases, spina bifida and schizophrenia in a single family associated with the heterozygote mutation of the heat-sensitive variant of methylenetetrahydrofolate reductase]. AB - Homozygous mutation of the thermolabile variant of methylene tetrahydrofolate reductase (MTHFR) may result in hyperhomocystinemia, leading to an increased risk for early cardiovascular disease, neural tube defects, and possibly major depression, schizophrenia. According to recent studies heterozygosity for the thermolabile variant of the MTHFR gene mutation is also more frequent in patients with thrombotic disease compared to that in the average population. We report on a family with different types of early vascular disease. In four consecutive generations MTHFR heterozygosity was detected: in the proband and in her mother, grandfather and daughter. Further conditions of the family members, possibly due to carrying the mutation, came to light by the pedigree analysis and examinations. The patient had pulmonary emboli at young age, her aunt died of spina bifida shortly after birth. The patient's mother suffers from schizophrenia and depression. The grandfather had pulmonary emboli, her sister with spina bifida occulta also carries the same mutation, as does her daughter who is sofar asymptomatic. In other asymptomatic members of the family no mutations were found. Unexpectedly, hyperhomocystinemia was detected in all heterozygote individuals. Our study demonstrates the necessity for folic acid therapy in mutation carriers to prevent early vascular events, depression and schizophrenia, and also to reduce the risk for neural tube defects in a preconception setting. PMID- 11481908 TI - [Plasma electrolytes in multiple myeloma]. AB - The plasma cell myeloma (multiple myeloma, myelomatosis) is a progressive disease, characterized by bone marrow plasmacytomas and the presence of monoclonal antibodies (IgG, IgA, IgD, IgE), or free kappa or lambda immunoglobulin side chains. The monoclonal antibodies or Bence-Jones protein may precipitate in the tubuli and impair kidney function. In addition, the plasma protein concentration may increase at the expense of plasma water level causing unrealistically low electrolyte levels. Since the isoelectric points of immunoglobulins are higher than those of most other plasma proteins, the net charge of plasma proteins may change causing new electrolyte balance. In addition, some monoclonal antibodies are more hydrated than others, and their high concentration may cause not only increased plasma viscosity but further electrolyte imbalance. In the present work the relationship between plasma protein and electrolyte levels is studied in samples of 100 multiple myeloma patients. PMID- 11481909 TI - [Adult-onset Kawasaki syndrome in the differential diagnosis of liver disease]. AB - The authors present a case of an adult with Kawasaki syndrome, who, due to jaundice, enlarged liver and abnormal liver function tests, was admitted the hospital with the suspicion of liver disease. The symptoms of Kawasaki syndrome appeared during the first nine days of the hospital stay. The authors emphasise, that liver function tests are frequently abnormal in adults presenting with this clinical entity, therefore Kawasaki syndrome should be taken into consideration in the differential diagnosis of liver diseases. PMID- 11481910 TI - [From smoking cessation to artificial ventilation. New therapy guidelines for COPD]. PMID- 11481911 TI - [Expanded arsenal for prevention and therapy. What is the value of the new malaria drugs? (interview by Waltraut Paukstadt)]. PMID- 11481912 TI - [Alcoholic and non-alcoholic fatty liver hepatitis. When is abstaining from alcohol enough, when do steroids help?]. AB - Alcoholic and non-alcoholic steatohepatitis (NASH) are often histopathologically indistinguishable. Apart from the clinical and laboratory findings, the patient's history is of key importance for differentiating between alcoholic steatohepatitis with its poor prognosis, and NASH, with its usually bland course. While the latter rarely requires specific treatment, the former, in particular in its severe form, is a therapeutic challenge. In addition to suitable dietary measures, patients with a Maddrey Score > 32, may need corticosteroids. In patients with mild to moderate steatohepatitis, the daily alcohol consumption appears to be decisive for the prognosis. In severe forms of this condition, in contrast, mortality is high, and patients surviving the acute phase often develop cirrhosis, irrespective of their further drinking habits. PMID- 11481913 TI - [Therapy of liver cirrhosis. Managing complications with fingertip control]. AB - Chronic liver diseases often lead to cirrhosis of that organ. As this progresses, hepatic function decreases, and the risk of life-threatening complications increases. Common complications are variceal bleeding, ascites, spontaneous bacterial peritonitis, hepatic encephalopathy, hepatorenal syndrome and hepatocellular carcinoma. The only therapeutic option that offers a chance of a cure is transplantation which, however, owing to strict selection criteria and the limited number of donor organs, can be applied only in a minority of patients. For most of the cases, the therapeutic strategy comprises treatment of the underlying disease, prevention and symptomatic treatment of typical complications. PMID- 11481914 TI - [Diabetics in the predialysis stage. Intensive therapy improves prognosis]. PMID- 11481915 TI - [PROGRESS Study examines transient cerebral ischemia and stroke patients. Future goal: attaining basic blood pressure control]. PMID- 11481916 TI - [Diabetics with digestive complaints. Is exocrine pancreatic function also impaired?]. PMID- 11481917 TI - [Only for limited indications. Biventricular pacemaker improves severe heart failure]. AB - In patients with congestive heart disease in whom left ventricular dilatation is associated with severely disordered contraction due to delayed ventricular conduction, a new form of treatment not requiring drugs is currently becoming established--biventricular pacing. Via an implanted electrode, biventricular stimulation accelerates conduction in the left ventricle and resynchronizes its contraction. Patients being considered for such therapy must have stable stage III disease (NYHA classification) under optimal medication, and a left bundle branch block with a QRS of at least 150 ms. Definitive recommendations with regard to indications will be possible only when current randomized mortality studies have been concluded. PMID- 11481918 TI - [Drug expenditures and health insurance contributions increase. Control of budgetsis at risk]. PMID- 11481919 TI - [Which antihypertensive drugs for diabetic patients? Remembering the organ protective potential!]. PMID- 11481920 TI - [Chronic inflammatory bowel diseases. Immunosuppressive drugs--good alternatives to steroids]. PMID- 11481921 TI - [Intermittent claudication. Walking plus vasoactive drug treatment]. PMID- 11481922 TI - [Rye pollen in prostatitis. Half the pain--double the pleasure in sex]. PMID- 11481923 TI - [ELSA Studies studies progression of atherosclerosis. Calcium antagonist arrests the process more than a beta-blocker]. PMID- 11481924 TI - [New potent serotonin receptor agonist. Helps migraine patients even when other triptans fail]. PMID- 11481925 TI - [20. Dysuria--typical symptoms lead to correct diagnosis]. PMID- 11481926 TI - The enhanced anticaries efficacy of a sodium fluoride and dicalcium phosphate dihydrate dentifrice in a dual-chambered tube. A 2-year caries clinical study on children in the United States of America. AB - PURPOSE: To clinically evaluate and compare a dentifrice system in a dual chambered tube, wherein one chamber contained sodium fluoride in a silica base and the other chamber contained dicalcium phosphate dihydrate (Test Dentifrice delivering 0.243% sodium fluoride), to a dentifrice containing 0.243% sodium fluoride in a silica base (Positive Control Dentifrice). MATERIALS AND METHODS: This study was conducted in harmony with the published 1988 American Dental Association guidelines for studies geared toward the comparison of fluoride dentifrices. This 2-yr caries clinical study employed a double-blind, parallel group design, and involved 5-17 yr-old children from the Central and South areas of Florida and from the Lares area of Puerto Rico. Qualifying subjects were stratified according to age and sex, and were randomly assigned to the two treatment groups, with multiple subjects in the same household all assigned to the dentifrice randomly allocated to the first among them. Caries examinations were conducted in accordance with U.S. Food and Drug Administration guidelines for the clinical evaluation of drugs to prevent dental caries. Two calibrated examiners performed all the measurements. After treatment assignment, study participants were instructed to brush their teeth at home with their assigned dentifrice at least twice daily. Brushing instructions were reinforced by indoctrination in proper oral hygiene techniques by dental professionals, supplemented by pamphlets supplied by the sponsor and yearly mailings to participants, emphasizing good oral hygiene and the need to enforce compliance with the study. Post-baseline examinations were performed after 1 yr of product use, and again after 2 yrs of product use. RESULTS: Two thousand five hundred six (2,506) subjects completed this 2-yr study. For these subjects, the mean caries scores (DMFS, decayed, missing and filled tooth surfaces) at baseline were 2.29 for the Test Dentifrice group, and 2.47 for the Positive Control Dentifrice group. For caries increments after 1 yr, the respective means were 0.69 for the Test Dentifrice group and 0.81 for the Positive Control Dentifrice group. Finally, after 2 yrs, the mean caries increments were 1.25 for the Test Dentifrice group, and 1.46 for the Positive Control Dentifrice group. No statistically significant difference was indicated between the treatment groups at baseline or between the 1-yr caries increment scores. However, there was a statistically significant difference in the 2-yr caries increment scores between the treatment groups. Relative to the Positive Control Dentifrice group, the Test Dentifrice group presented a 14.38% reduction in caries increment scores at 2 yrs. In accordance with the procedures and standards provided by the published guidelines of the American Dental Association for the comparison of the anticaries efficacy of fluoride dentifrices, the results of this study support the conclusion that the dentifrice system in a dual-chambered tube, wherein one chamber contained sodium fluoride in a silica base and the other chamber contained dicalcium phosphate dihydrate, delivering 0.243% sodium fluoride, provided a superior level of anticaries efficacy than did the dentifrice containing 0.243% sodium fluoride in a silica base. PMID- 11481927 TI - The enhanced anticaries efficacy of a sodium fluoride and dicalcium phosphate dihydrate dentifrice in a dual-chambered tube. A 2-year caries clinical study on children in Brazil. AB - PURPOSE: To clinically evaluate and compare a dentifrice system in a dual chambered tube, wherein one chamber contained sodium fluoride in a silica base and the other chamber contained dicalcium phosphate dihydrate (Test Dentifrice delivering 0.243% sodium fluoride), to a dentifrice containing 0.243% sodium fluoride in a silica base (Positive Control Dentifrice). MATERIALS AND METHODS: This study was conducted in harmony with the published 1988 American Dental Association guidelines for studies geared toward the comparison of fluoride dentifrices. This 2-yr caries clinical study employed a double-blind, parallel group design, and involved 6-10 yr-old children from the metropolitan area of Maceio, Alagoas, Brazil. Qualifying subjects were stratified according to age and sex, and were randomly assigned to the two treatment groups, with multiple subjects in the same household all assigned to the dentifrice randomly allocated to the first among them. Caries examinations were conducted in accordance with U.S. Food and Drug Administration guidelines for the clinical evaluation of drugs to prevent dental caries. One calibrated examiner performed all the measurements. After treatment assignment, study participants were instructed to brush their teeth at home with their assigned dentifrice at least twice daily. Brushing instructions were reinforced by indoctrination in proper oral hygiene techniques by dental professionals, supplemented by pamphlets supplied by the sponsor and yearly mailings to participants, emphasizing good oral hygiene and the need to enforce compliance with the study. Post-baseline examinations were performed after 1 yr of product use, and again after 2 yrs of product use. RESULTS: Two thousand four hundred thirty-two (2,432) subjects completed this 2-yr study. For these subjects, the mean caries scores (DMFS, decayed, missing and filled tooth surfaces) at baseline were 3.84 for the Test Dentifrice group, and 4.06 for the Positive Control Dentifrice group. For caries increments after 1 yr, the respective means were 2.02 for the Test Dentifrice group and 2.12 for the Positive Control Dentifrice group. Finally, after 2 yrs, the mean caries increments were 4.30 for the Test Dentifrice group, and 4.83 for the Positive Control Dentifrice group. No statistically significant difference was indicated between the treatment groups at baseline or between the 1-yr caries increment scores. However, there was a statistically significant difference in the 2-yr caries increment scores between the treatment groups. Relative to the Positive Control Dentifrice group, the Test Dentifrice group presented a 10.97% reduction in caries increment scores at 2 yrs. In accordance with the procedures and standards provided by the published guidelines of the American Dental Association for the comparison of the anticaries efficacy of fluoride dentifrices, the results of this study support the conclusion that the dentifrice system in a dual chambered tube, wherein one chamber contained sodium fluoride in a silica base and the other chamber contained dicalcium phosphate dihydrate, delivering 0.243% sodium fluoride, provided a superior level of anticaries efficacy than did the dentifrice containing 0.243% sodium fluoride in a silica base. PMID- 11481928 TI - Development of an enhanced anticaries efficacy dual component dentifrice containing sodium fluoride and dicalcium phosphate dihydrate. AB - A dual-chamber dentifrice, which contains sodium fluoride (NaF) in one component and dicalcium phosphate dihydrate (dical) in the other, has been developed. The dentifrice is packaged in a dual-chamber tube and is formulated to deliver 1100 ppm F. A series of studies consisting of in vitro fluoride uptake, in vivo calcium labeling, intraoral remineralization-demineralization, and animal caries studies were performed to support the improved anticaries efficacy of this product in comparison to a sodium fluoride/silica dentifrice (NaF/silica). An in vitro fluoride uptake study comparing 1100 ppm F NaF/dical dentifrice to 1100 ppm F NaF/silica showed that NaF/dical delivered significantly more fluoride than NaF/silica, 3.72 +/- 0.36 micrograms/cm2 versus 2.41 +/- 0.10 micrograms/cm2. A 6 day in vivo brushing study with a 44Ca labeled NaF/dical dentifrice showed that calcium from dical penetrated demineralized enamel and was present in plaque up to 18 hrs since the last brushing. An intra-oral remineralization demineralization study was performed to evaluate NaF/dical's ability to promote remineralization in comparison to three silica-based dentifrices containing 0, 250, and 1100 ppm F as NaF. The percent mineral changes after treatment were +20.44 +/- 17.14 for NaF/dical, +9.27 +/- 19.53 for 1100 ppm NaF/silica, -1.43 +/ 20.57 for 250 ppm NaF/silica, and -12.36 +/- 32.76 for 0 ppm F/silica. A statistical analysis showed that the dual-chamber NaF/dical dentifrice was significantly more effective than the 1100 ppm NaF/silica dentifrice at promoting remineralization. A rat caries study was performed to evaluate NaF/dical ability to prevent caries in comparison to 1100 ppm F NaF/silica, 250 ppm F NaF/silica, silica, and dical dentifrices. The mean smooth surface caries scores were 1.6 +/- 2.8 for NaF/dical, 5.5 +/- 6.2 for 1100 ppm F NaF/silica, 10.6 +/- 6.2 for 250 ppm F NaF/silica, 13.7 +/- 4.7 for 0 ppm F/silica, and 9.5 +/- 7.8 0 ppm F/dical. A statistical analysis showed that the the dual-chamber NaF/dical dentifrice was superior to all other treatments tested in preventing caries in rats. The dical dentifrice was significantly superior to the silica dentifrice in preventing caries, which indicates that dical alone exhibits anticaries efficacy. In conclusion, individual and cumulative results from the fluoride uptake, intra oral remineralization-demineralization, and rat caries studies from the dual chamber NaF/dical dentifrice support the improved anticaries efficacy of this product. PMID- 11481929 TI - Don't burn out ... blaze new trails! PMID- 11481930 TI - Online learning. Now playing at a computer near you. PMID- 11481931 TI - Can advertising help in the race to success? PMID- 11481932 TI - When I grow up.... I wanna be a dentist. PMID- 11481933 TI - Volunteer ensures health care for the uninsured. PMID- 11481934 TI - TMA. 2000 annual report and resource guide. PMID- 11481935 TI - Richard G. Popiel, MD. Vice president and chief medical officer of Horizon Blue Cross Blue Shield of New Jersey. PMID- 11481936 TI - West Nile virus: New Jersey's 2000 experience and surveillance plans for 2001. PMID- 11481937 TI - Medicare paperwork abuses doctors, harms patients. PMID- 11481938 TI - Paterson, New Jersey, and William Carlos Williams, MD. PMID- 11481939 TI - Professional liability insurance: II. The legal environment. PMID- 11481940 TI - Knowledge of stroke risk factors, symptoms, and treatment among New Jersey adults. PMID- 11481941 TI - Implementing a policy and procedure manual. PMID- 11481942 TI - Proponent of evidence-based medicine joins Horizon Blue Cross Blue Shield. PMID- 11481943 TI - West Nile virus 2000-2001. PMID- 11481944 TI - Making sense of pain. PMID- 11481945 TI - A world of pain treatment options. PMID- 11481946 TI - Pain in perspective. PMID- 11481947 TI - The future. Will pain be abolished or just pain specialists? PMID- 11481948 TI - Chronic noncancer pain management. PMID- 11481949 TI - Cancer pain management. AB - The management of cancer-related pain is often quite different from the management of chronic benign pain (see companion article by Thomas Elliott, M.D., page 28). As cancer therapy becomes increasingly complex, the management of pain and other symptoms requires more innovative approaches as well. This article begins with a basic overview of how to evaluate the patient with cancer pain, offers simple opiate use guidelines, and discusses newer agents for neuropathic and somatic pain. Finally, it discusses how future pain management strategies may hinge on research and cites examples of studies being performed in Minnesota. PMID- 11481950 TI - Teens and tobacco in Minnesota. New findings from the Minnesota youth tobacco survey. AB - The Minnesota Youth Tobacco Survey (MYTS) was administered to 12,376 public school students in grades 6 through 12 to obtain baseline data at the start of the state's latest efforts to reduce tobacco use among youth. The sample is representative of public school students in the state. More than one-third of high school students (38.7%) and 12.6% of middle school students are current tobacco users, defined as use of tobacco on one or more days in the previous 30 days. Current use of cigarettes is 32.4% in high school and 9.1% in middle school. More than 90% of students, including most smokers, agree that tobacco use is addictive and harmful. More than three-fourths (78.4%) of current smokers report that two or more of their four closest friends are smokers. Students who live with a smoker are more likely to become current smokers than students who do not live with anyone who smokes (42.5% to 25.1% in high school). Most current smokers (61.0%) tried to quit smoking at least once in the past 12 months. Eighty eight percent (88.4%) of current smokers in high school smoke one of the top three advertised brands: Marlboro, Camel, or Newport. Results of the MYTS illustrate some of the challenges facing the state's tobacco reduction efforts and some of the factors that must be considered as Minnesota tries to design and implement effective programs. PMID- 11481951 TI - Managing a mystery. PMID- 11481952 TI - [Contingency of the national hospitalization system for peace time emergencies]. PMID- 11481953 TI - [Ethical considerations in mass casualty situation]. AB - Mass casualty is a situation, in which, the physician is compelled to make critical decisions under heavy pressure load, due to severe shortage in time, personnel and information. This task is extremely difficult to fulfill, as the physician has to consider not only professional tools, but needs also to utilize ethical principles, in order to provide the best possible care to most of the casualties who might benefit from it. By definition, in the mass casualty situation the medical facility lacks temporarily the ability to deliver effective therapy to all, as the injured outnumber the medical capacity for a given time. The ethical conflicts and dilemmas that arise during such an event are enormous. Amazingly, only a few articles have addressed the issue of ethical considerations during mass casualty situation. Ethical decision making is based on four principles: beneficence, nonmaleficence, autonomy and justice. Compassion, trustworthiness, discernment and integrity are the four qualities required from those practicing medicine. These virtues should be manifested in mass casualty situations, one of the most demanding situations a physician may face. PMID- 11481954 TI - [Liver failure with coagulopathy in an infant with tyrosinemia]. AB - Tyrosinemia is an inherited autosomal recessive condition. We present a 5 week old boy with this disorder. He was admitted because of a fever, vomiting and lethargy. The laboratory tests confirmed a coagulopathy with prolonged prothrombin time (PT), partial thromboplastin time (PTT) and a decreased serum fibrinogen. The alpha-fetoprotein level was markedly elevated. To confirm the diagnosis of tyrosinemia, quantitative urinary succinylacetone was measured. Although overt liver failure with coagulopathy may be part of the representation of tyrosinemia, a significant coagulopathy in the absence of overt signs of liver disease has not been emphasized as a clue to the diagnosis of this condition. PMID- 11481955 TI - [Costs and benefits of laparoscopic inguinal hernia repair--is there an economic justification?]. AB - BACKGROUND: With the recent accelerated development of laparoscopic surgery, it has been applied in all fields of surgery. The main issue today is not the technical ability of performing laparoscopic procedures but rather their justification while considering the disease and cost-benefit aspects. OBJECTIVES: The present study surveys the experience accumulated in recent years concerning the economical aspects of laparoscopic inguinal hernia repair. Issues like length of the surgical procedure, cost, typical complications and recuperation time are being addressed. METHODS: We reviewed controlled clinical trials that compare laparoscopic and open repair of inguinal hernia. Trials that included at least 100 patients and were published in peer-reviewed journals since 1996 were included. Various operative techniques were compared using clinical and economical parameters. RESULTS: Most studies examined the issues from an institutional or health care system viewpoint and not from a comprehensive societal perspective. Therefore, indirect costs were seldom included. Operative times were longer and direct costs were significantly higher in the laparoscopic approach as compared to conventional open surgery. However, recovery time and return to work were found to occur earlier in patients who underwent laparoscopic surgery. CONCLUSIONS: A shorter recovery time and shorter off-work period after laparoscopic hernia repair could compensate for the increased hospital expenditures. However, this contribution has not yet been completely established and needs further validation by supplementary studies. Other aspects that will have to be evaluated include quality of life and patient satisfaction. PMID- 11481956 TI - [Menetrier's disease in children]. AB - Menetrier's disease in children is a rare disorder that is characterized by the presence of marked protein losing gastropathy associated with enlarged and thickened gastric folds. Abnormal regulation of gastric epithelial growth, probably triggered by an infectious agent, has been suggested as an etiology for this disorder. We describe a case of Menetrier's disease in a young child and review the current literature encompassing the different aspects of the disease. PMID- 11481957 TI - [Male genital self mutilation--a cause of hematuria]. AB - Pathological self-mutilation is defined as a deliberate destruction of body tissue without conscious intent. It is sometimes associated with several pathologies such as mental disorders and mental retardation. We report a non psychotic male, with borderline mental retardation who practiced sporadic moderate genital self-mutilations as a masturbation technique and presented as a case of recurrent gross hematuria. PMID- 11481958 TI - [Inflammatory bowel disease and osteoporosis--an evidence-based connection]. AB - Inflammatory bowel diseases (IBD) often have chronic or remittent course. Control of symptoms and disease activity often requires the administration of glucocorticoids. Osteoporosis is a frequent finding in patients with IBD. The evidence-based connection between the two conditions relies on a relatively limited number of cross-sectional and longitudinal studies. The cause of this connection has not been defined completely. Probably, both the chronic inflammation and the steroid therapy contribute to the osteopenia, through an increase in bone resorption without a compensatory increase in bone formation. There is little data regarding the efficacy of therapeutic interventions for the prevention of osteoporosis in patients with IBD. Recently, guidelines for the evaluation and treatment of osteoporosis and IBD have been published. There is need for further research on the relation between these conditions and on the efficacy of current therapy for this unique population. PMID- 11481959 TI - [Prenatal Down's syndrome screening at 10-14 weeks gestation using the combined nuchal translucency and maternal serum biochemistry: preliminary results of the first 358 cases]. AB - We report our preliminary experience of prenatal screening for Down's syndrome (DS) using nuchal translucency (NT) measurement combined with the serum biochemistry analysis of Free beta-human chorionic gonadotropin (F beta hCG) and pregnancy associated plasma protein A (PAPP-A) all measurement at 10-14 weeks of gestation. Of the 358 parturient women which enrolled in the study, 9 cases were not included because of fetal anomalies or miscarriages. Thus the study group included 349 singleton pregnancies in which complete prenatal and infant follow up was available. Forty-four pregnant women were found to be screen positive (12.6%) and in 13 cases (27%) of them fetal chromosomal aneuploidies were diagnosed. Looking into the markers profile we found that the NT was a sensitive marker which was abnormally increased in all the fetal aneuploidies. Serum F beta hCG was found to be a promising marker as well, being significantly elevated (2.26 +/- 0.86 multiple of the medians, MoM) in DS cases, and decreased (< 0.5 MoM) in two cases of Edward's syndrome. On the contrary, PAPP-A was found less sensitive, and its mean MoM values were not significantly different between DS versus euploid fetuses. Our preliminary results support the promising success of DS screening using NT and F beta hCG. PMID- 11481960 TI - [Nephrogenic adenoma--benign lesion of urinary tract system]. AB - Nephrogenic adenoma is a rare metaplastic benign lesion of the urinary tract caused by chronic irritation to the urinary epithelium. The appearance of this lesion is usually characterized by hematuria and irritative symptoms. Nephrogenic adenoma may be found most commonly in the bladder and the urethra and less frequently in the renal pelvis and ureters. We present 5 patients who underwent surgery due to bladder or urethral nephrogenic adenoma. Despite the fact that nephrogenic adenoma is a benign lesion, long term follow-up is needed due to the high recurrence rate and the potential, though rare, malignant transformation. PMID- 11481961 TI - [Hospital-at-home as a solution for the treatment requirements of acute exacerbation in multiple sclerosis]. AB - Cooperation between the Multiple Sclerosis center at the Carmel medical center and the Hospital-at-Home (H.H) department of the continuing care unit in the Haifa and Western Galilee district of the Clalit Health Services has made it possible to give methylprednisolone intravenously to Multiple Sclerosis (M.S) patients during an acute exacerbation of the disease, in their home. In this study, we describe the joint work of the two centers. We have summarized 30 treatment courses given to 26 patients in their homes, following referral by the M.S. center, in the year 1999. The aims of the study included assessing satisfaction, safety and cost-effectiveness in a treatment course in the HH framework, as compared to the same treatment being conducted in the framework of hospitalization in various neurological departments, as was done in the past in the same group of patients. The expenses involved in HH for this group of patients were only 14% of the parallel treatment in the hospital (a savings of 86%). The treatment has proven to be extremely safe. There were no side-effects that required returning patients to the hospital, and the treatment was given in conditions of maximum comfort for the patient and his family. A telephone survey was conducted, which compared the satisfaction with the HH treatment, and the burden caused the patient's family to prior hospitalization for the same treatment. For all of the parameters examined, greater satisfaction was distinctly proven in the HH treatment. In light of these findings, we can conclude that giving methylprednisolone intravenously to M.S patients during an acute exacerbation, in the HH framework, is a safe and cost effective treatment, preferred by the patient and his family. PMID- 11481962 TI - [Myocardial infarction due to cocaine]. AB - Cocaine is a common drug. Myocardial infarcts and brain infarcts related to the use of cocaine were previously reported in the medical literature. We report a 34 year old patient admitted for severe chest pain, a few minutes after cocaine use. He was found to have an acute myocardial infarction with significant left ventricle dysfunction. Coronarography conducted a few days later was found to be normal. Cocaine use may cause acute myocardial infarction in young and healthy people with no other risk factors. PMID- 11481963 TI - [The many faces of syringomyelia]. AB - During July-August 1998, we encountered 4 patients who suffered from syringomyelia. The diagnosis in each case could not be reached on clinical grounds only, since the characteristic thermoanalgesia dissociation was absent in all patients. In each case the spinal MRI showed the typical intraspinal cystic lesion, which finally established the diagnosis and led to appropriate treatment. PMID- 11481965 TI - [Modern radiology in Israel--a "catch" situation]. PMID- 11481964 TI - [The interaction between saliva and cigarette smoke and its devastating biological effects as related to oral cancer]. AB - Saliva is the first body fluid to confront inhaled cigarette smoke (CS) which is injurious to the oral cavity and is associated with several oral diseases and cancer. We have recently demonstrated in an In vitro model that an exposure of saliva to nine 'puffs' of CS induced a distinct increase in protein carbonyls. This elevation of protein carbonyls caused by CS may be the result of aldehydes presented in the CS reacting with sulphydryl (-SH) groups of the proteins. Moreover, following the exposure to CS, the activities of several salivary enzymes amylase, lactate dehydrogenase (LDH) and acid phosphatase (ACP) were found to be significantly reduced by 83%, 57% and 77%, respectively. However, CS had no effect on the activities of aspartate aminotransferase and alkaline phosphatase. In the current study we found that CS also reduced salivary peroxidase activity by no less than 80% which may be of a great importance to the clinical set up as peroxidase is considered a pivotal enzyme of the the salivary protecting system. Furthermore, in contrast to LDH findings in saliva, the LDH activity in plasma was not reduced following its exposure to CS. CONCLUSION: Hence, we concluded that the loss of salivary enzyme activities may be due to various agents in the CS that affect the enzyme via different mechanisms. PMID- 11481966 TI - [Are atherosclerotic process involved in aortic-valve calcification?]. PMID- 11481967 TI - [Trends of medical research in Israel]. PMID- 11481968 TI - [The revolution of stem cells: bone marrow cells, neural cells and muscle cells are intercovertible]. AB - For many years, the fate of adult cells has been assumed to be limited to their tissues of origin. Upon development cells become increasingly specialized to carry out particular functions. Recently, numerous studies have shown that some stem cells possess greater plasticity than previously envisioned. Thus, a few donor bone-marrow cells when transfused into immunodeficient mice, were recovered as glial cells in the host brain, or as cells expressing neuronal antigens. Alternatively, neural stem cells could differentiate into myeloid and lymphoid cell lineages upon transplantation into the hematopoietic system of irradiated hosts. PMID- 11481969 TI - [Diabetic dyslipidemia]. PMID- 11481970 TI - [Embryo transfer and multiple gestation--review of issues influencing the number of embryos transferred in in vitro fertilization and embryo transfer]. PMID- 11481971 TI - [Massive pulmonary embolism: an update on management]. PMID- 11481972 TI - [Dopamine in acute renal failure]. AB - Dopamine is widely used for prevention and treatment of acute renal failure. Its application is based on physiological data suggesting a selective renal effect when it is administrated at a low dose. There is no evidence that dopamine is useful in preventing acute renal failure in patients undergoing high risk procedures. Most of the studies demonstrated lack of benefit from dopamine treatment in cases of acute renal failure. Dopamine has a number of known side effects. Large randomized controlled trials are required in order to examine outcomes such as mortality, the need for dialysis or the length of hospitalization. CONCLUSION: The use of dopamine for prevention and treatment of acute renal failure should be avoided. PMID- 11481973 TI - [Possible relevance of alterations at the cellular, neuronal connectivity and brain structure levels to the pathophysiology of schizophrenia]. PMID- 11481974 TI - [Thrombotic thrombocytopenic purpura--new insights in the pathogenesis, diagnosis and modern therapy]. PMID- 11481975 TI - [On the history of the ambulance]. PMID- 11481977 TI - [Impressions and lessons from the National Forum for Quality in Health--San Francisco, USA December 2000]. PMID- 11481976 TI - [A summary on the first convention of the European Committee for Palliative Medicine--Berlin, Germany December 2000]. PMID- 11481978 TI - [The thalidomide story and its lessons]. PMID- 11481979 TI - [Moderate alcohol drinking and coronary heart disease--what are the most prudent recommendations for the public?]. AB - The author reviews some epidemiological evidence for the association between moderate alcohol drinking and lower risk for coronary heart disease (CHD). The argument being that the epidemiological evidence does not necessarily mean that alcohol itself is responsible for the lower risk, and describes lifestyle factors that might have influence on the association and make alcohol's role incidental and not causal. In addition, it illustrates the reasons for the fact that the "French Paradox" cannot be explained solely by alcohol use, and emphasizes that there is no consensus to say that alcohol consumption in moderation is protective or that a certain type of alcoholic beverage is protective. However, in case alcohol has some role in the inhibition of the development of CHD, the author describes possible biological mechanisms that might suggest paths to explain the apparent protective effects of alcohol. The article deals with the balance between alcohol's beneficial and harmful effects on health and provides recommendations for the public concerning alcohol use and CHD. PMID- 11481980 TI - [The role of posterior release in the treatment program for congenital clubfoot]. AB - The aim of this paper was to assess the utility of this procedure and to define it's role in treatment of congenital clubfeet. The material comprises 123 children, among whom 154 feet were treated by posterior release. The age of the patients ranged from 5 to 36 months (average age: 16 months). The procedure involved the lengthening of the Achilles tendon in the sagittal plane, partial resection or transverse dissection of the articular capsule of the ankle joint. In selected cases lengthening of the tendon of the extensor hallucis muscle was performed and sometimes of the posticus muscle. The described procedure was performed in all cases where all forefoot components of the deformity were found, as well as an equines position of the foot. A group of 87 patients (70.7%), among whom posterior release was performed in 101 feet (65.6%). The follow-up time ranged from 5 to 15 years (average: 12.3 years). Results were assessed according to the classification by Magone et al. Very good results (95-100 points) were achieved in 27 feet (26.8%), good results (80-89 points) were found in 40 feet (39.6%), satisfactory (70-79 points) in 25 feet (24.7%), and bad results (less than 70 points) in 9 feet (8.9%). Basing on their own experience the authors' believe this procedure is very useful in treatment of congenital clubfeet. It's therapeutic usefulness is at it's best when deformities of the forefoot have been conservatively corrected. The extent of posterior release depends on the severity of the deformity. PMID- 11481981 TI - [Use of pedicled radial forearm flaps for covering of extensive soft tissue defects in the upper extremity]. AB - Results of treatment of 13 patients with extensive defects of soft tissues in the upper extremities treated with pedicled radial forearm flaps are presented (11 fascio-cutaneous flaps and 2 adiposo-cutaneous). The group included 5 females and 8 males aged 9-85 years (mean age: 37 years). The defects followed trauma in 12 cases and in one case it was the result of oncological resection. The flap was used as a primary procedure in 4 cases after amputation of the finger or degloving of the hand. The flap appeared very useful in delayed reconstructions in older patients with extensive defects of soft tissues and bones, especially in the elbow region. Complete survival of all flaps with very good late results were obtained. Donor sites were covered with skin grafts and healed correctly in 11 patients. In two patients healing of the donor site prolonged in time for over 6 weeks. No other complications were observed using this method. PMID- 11481982 TI - [Corrective French's osteotomy in treatment of cubitus varus]. AB - Cubitus varus is the most common late complication of supracondylar fracture of the humerus in children. The normal carrying angle can be restored by supracondylar osteotomy. This operation using the technique described by French. was done in 10 patients. During follow up examination motion of the cubitus was assessed and compared with an uninjured side. The x-ray examination included evaluation of the carrying angle. Favourite results were confirmed by restoration of carrying angle with minor loss of mobility of the elbow joint with no recurrence of the varus deformity during follow up. PMID- 11481983 TI - [Proximal femur remodeling after proximal femur varus-derotational osteotomy in treatment of developmental dysplasia of the hip]. AB - The aim of this paper was to assess proximal femur remodelling after proximal femur varus-derotational osteotomy in treatment of developmental dysplasia of the hip up to skeletal maturity. The analysis included 48 osteotomies performed in 33 children between 1st and 10th year of life. The follow-up period was on average 12 years. Supported by statistical evaluation the analysis showed persistent increase of the neck-shaft angle after proximal femur varus-derotational osteotomy, especially in the younger age groups. It also correlated strongly with the amount of varus fixed at the time of operation. On other hand femoral antetorsion corrected during surgery remained unchanged till skeletal maturity. PMID- 11481984 TI - [Evaluation of localization and size of subcapital growth plate damage of the proximal femur in the MR image]. AB - A group of 33 hips with avascular necrosis following developmental dislocation of the hip in 24 children was evaluated. The average age of the patients at the time of evaluation was 10 years (age ranging from 4 to 14 years). The study was focused on depiction of the subcapital growth plate of the femur by means of conventional radiography and MRI. On the basis of of the analysis of MR imagesa method of mapping the subcapital growth plate allowing location of the damage site is presented. The method will be useful in planning corrective treatment of the proximal femur e.g. trochanteric epiphysiodesis, corrective osteotomy and future removal of the bone bridges. PMID- 11481985 TI - [Corrective surgical procedures as repeated treatment of residual clubfoot]. AB - The authors present an analysis of failed surgery in 139 patients with congenital clubfeet. There were 72 feet, which required repeated surgical procedures after primary treatment. Twelve feet were treated by posterior release, and 65 by postero-medial release. The repeated reconstruction was performed at a mean age of 3.6 years. No very good results were found in either group at final follow-up, done according to the Magone et al. classification. In the first group treated by posterior release god results were noted in 53.3% of the cases, sufficient in 43.1% and poor in 38.5%. The authors underline that the final results depend on early consequent conservative treatment, use of adequate operating techniques and cooperation between child, parents, physician and orthotic specialist. PMID- 11481986 TI - [Evaluation of methotrexate levels in biological fluids released from bone cement filling]. AB - The authors present an analysis of methotrexate concentration in blood, urine and drainage liquid (both deep and superficial), as well as the results of biochemical analysis of blood and urine in a population of 42 patients treated surgically because of pathologic fractures of long bones. In 8 of the 42 patients chemotherapy was applied 14 days before the surgical procedure. Lithic lesions in the long bones were filled with cement with 2 g of methotrexate and further stabilisation was achieved by applying metal implants. The highest methotrexate concentrations were noted in deep drainage fluid and in blood. Methotrexate concentration reached its peak in body fluids 12-24 hours after the procedure, and fell to a minimum 48-72 hours post surgery. The lowest level of the drug compared to all body fluids was found to be in the blood. Only after 72 hours did it exceed the minimum required level of 0.05 umol/l. All biochemical markers in blood and urine except AspAT were found within normal range in all cases 72 hours post surgery. After 24-72 hours post-op a decrease in the number morphotic elements of blood was noted, particularly lymphocytes and thrombocytes. PMID- 11481987 TI - [Evaluation of treatment results for osteochondritis dissecans of the knee joint]. AB - The paper presents the results of treatment of 66 patients with osteochondritis dissecans of the knee in the period between 1987 and 1998. Arthroscopically assisted removal of loose bodies was performed in 14 patients, arthroscopy and fixation of the lesion with one or two screws was performed in 5 patients and arthroscopically assisted drilling of the site of the defect was performed in 26 patients. The patients were clinically evaluated according to the 100-point Lysholm scale. Basing on the Lysholm scale, results wee classified as excellent in 11 patients, as good in 15 patients, fair in 14 patients and poor in 5 cases. Treatment of osteochondritis dissecans by excision of the loose or partially attached fragment, with shaving of the base of the crater does not give good results. Excision should be reserved for small fragments that cannot be replaced anatomically. Restoration of congruity of the joint surfaces by fixation of the lesion fragment prevents future gonarthrosis and gives good final results. PMID- 11481988 TI - [Use of radiotherapy in treatment of painful scapula-humeral periarthritis]. AB - In 10 cases of painful scapulo-humeral periarthritis radiotherapy was used. In all cases conventional anti-inflammatory treatment was unsuccessful or impossible to deliver. Patients were treated using gamma 60Co radiation and 6 Gy was delivered in 1 Gy fractions over 8 days. The average degree of pain relief and improvement of limb mobility was assessed after radiotherapy completion, 1 week, 7 weeks after treatment and at final-up. The degree of pain relief was respectively 27%, 72%, 85% and 93%. Significant improvement of limb mobility was noted in all cases. Obtained results allow to conclude that anti-inflammatory radiotherapy is an effective treatment of painful scapulo-humeral periarthritis. PMID- 11481989 TI - [Computer analysis of bones and regeneration on radiographs taken during limb lengthening--introductory comments]. AB - Computer assisted image analysis is used for qualitative assessment of radiographs more and more often. The aim of this study was to define a method enabling objective assessment of changes in bone structure, using standard radiographs during limb elongation. Application of such analysis for assessment of bone and regenerate during limb elongation is described. The method of estimation of the obtained results is also presented. The obtained results encourage the authors to continue their studies, so as to make objective assessment of bone using standard radiographs a common method. PMID- 11481990 TI - [Osteochondrodysplasia determined genetically by a collagen type II gene mutation]. AB - Chondrodysplasias are a heterogenous group of skeletal dysplasias, affecting the growing cartilage. The main part of chondrodysplasias is caused by mutations in various types of collagen genes. The current classification within this group of disorder relies on clinical, histological and radiographic features. Type II collagenopathies comprise part of chondrodysplasias, consisting of hereditary disorders caused by defects in the type II collagen. Collagen type II is coded by a large gene--COL2A1. The chromosomal location for the human COL2A1 gene is 12q13.11-q13.12. Defects in collagen type II are caused by point mutations in the COL2A1 gene. Type II collagenopathies form a wide spectrum of clinical severity ranging from lethal achondrogenesis type II, hypochondrogenesis, through severe forms like spondyloepiphyseal dysplasia congenita, spondyloepimetaphyseal dysplasia congenita, Marshall syndrome, to the mild forms--Stickler syndrome and early osteoarthritis. The pathological changes in the patients are observed in the growth plate, nucleus pulposus and vitreous body, where the abnormal collagen type II is distributed. This article presents the genetic background of collagenopathies type II and the results of current molecular studies of the patients. Both the molecular and the clinical studies may promise a better understanding of the relationship between the genotype and the phenotype. We present the patients, who were diagnosed at the Department of Medical Genetics and in the Orthopaedic Department in Poznan. PMID- 11481991 TI - [Endoprostheses for one hip and both knee joints in a patient with rheumatoid arthritis after supracondylar fractures of both femurs]. AB - The paper presents a case of hip prosthesis and ambilateral knee prosthesis after ambilateral supracondylar fractures of the femur healed in malposition in a patient with rheumatoid arthritis. The knee prosthesis were implanted without application of the femoral intramedullare rod. Three prosthesis were successfully implanted over a period of 8 weeks, obtaining pain relief and satisfactory range of movement. PMID- 11481992 TI - [Nonsurgical treatment of patients with cervical spine pain--algorithm for proceeding]. AB - Diagnosis and treatment of neck pain still remains a therapeutic challenge. Basing on available literature and their own experience the authors present an algorithm for the management of neck pain in daily orthopaedic practice. It is crucial to correlate correctly the patient's complaints, the physical examination and the laboratory findings (including conventional imaging modes and neurodiagnostic procedures). The chronologically organised algorithm should help the physician in find the optimal therapeutic solution for his patient. The use of this algorithm should also lead to a decrease in the number of unnecessarily performed surgical procedures. PMID- 11481993 TI - The politics and policies of caring for the elderly in an age of retrenchment: a symposium, Part 2. PMID- 11481994 TI - The interests of three stakeholders in independent personal care for disabled elders. AB - There is concern throughout developed countries about how sufficient services can be mounted in this century to care for all older, disabled persons who want to remain at home. With declining availability of family care and high turnover among paid workers, cost, care supply, and quality dilemmas abound. Therefore, how can home care be improved? One suggestion is to revise the care "contract" by simply eliminating the costly bureaucratic and controlling dimensions of agency oversight and management. In consumer-directed or self-managed care, independent workers are hired directly by consumers or their family members. Consumers spend their own private money or government provides authorization for them to "hire" their own workers. Such arrangements may be more responsive to the needs of consumers, more directly accountable to them, and less costly for both individuals and government. This article examines 41 cases of consumer-directed care in Milwaukee. Extended interviews with three "stakeholders" in care--the disabled, older client, the family caregiver, and the personal assistant--were conducted, including both private pay and government subsidized arrangements. This analysis identifies some essential, but disparate interests of the three partners in care. PMID- 11481995 TI - The demographic, economic, and health profile of older Latinos: implications for health and long-term care policy and the Latino family. AB - The number of older Latinos is increasing rapidly and many of its members are in poor health, economically disadvantaged, and reliant on publicly-funded programs that are at risk of continuing cutbacks and restructuring. Such changes limit this population's access to health and long-term care services and shift an increasing amount of responsibility to the Latino family. This article reviews the demographic, economic, and health data of the Latino population and also discusses current and proposed changes under Medicare and Medicaid and the potential impact of these revisions on the Latino population. The authors argue that the needs of younger and older Latinos are linked and any program and policy to improve the situation of the elderly must confront the vulnerabilities of all family members. PMID- 11481996 TI - Issues confronting lesbian and gay elders: the challenge for health and human services providers. AB - During the past two decades, the number of people over the age of 65 in the United States has been increasing twice as fast as the rest of the population. Within this diverse group, gerontologists are increasingly aware that there also exists a large population of older lesbian, gay men, and bisexuals. Health professionals must learn to acknowledge and meet the needs of this population. This article assists in addressing the knowledge gap by acquainting the reader with the aging concerns of lesbian and gay men; the impact of homophobia on their health; common heterosexual practices and their impact on care; retirement and leisure issues; the hidden incidence of abuse and neglect; and some suggested strategies that will assist health and human services workers in providing quality care for lesbians and gays as they age. PMID- 11481997 TI - Aging in America: challenges and strategies for health care delivery. AB - This article delineates the current and future trends related to aging in the United States. The demographic and economic forces outlined are ones that will clearly challenge health services leaders and policy-makers in the coming decades. Various policy interventions and strategies currently underway to address the needs of the aging population are described. Shifts in caregiving patterns, the delivery of long-term care services, economics, choices in living arrangements, and managed care programs for the elderly are a few of the issues presented in this article. Health and human services for the elderly will undergo a profound change as the health system continues to adapt itself to the demands of an aging society. PMID- 11481998 TI - Redesign in the long-term care industry: a restraint reduction or restraint elimination program in the nursing home. AB - The Nursing Home Reform Act of 1987 required nursing homes in the United States to modify their use of physical and chemical restraints in the provision of care to the residents. Nursing home staff members can restrain only if a medical professional determines that they must temporarily restrain residents in order to provide care for them. The author employs the Kast and Rosenzweig model of systems theory to describe the approaches that nursing homes have taken to implement a restraint-free or restraint-elimination program. He then describes the benefits that residents and staff members receive when they participate in a restraint-reduction of restraint-elimination program. PMID- 11481999 TI - Amiodarone induced thyroid dysfunction: pathophysiology, diagnosis and management. PMID- 11482000 TI - Hertfordshire medieval hospitals. AB - All Hertfordshire's hospitals were small, although some had considerable endowments, e.g. St Julian's Hospital at St Albans. Most were in decline by the late 15th Century, or had ceased to function. PMID- 11482001 TI - Adverse and beneficial effects of plant extracts on skin and skin disorders. AB - Plants are of relevance to dermatology for both their adverse and beneficial effects on skin and skin disorders respectively. Virtually all cultures worldwide have relied historically, or continue to rely on medicinal plants for primary health care. Approximately one-third of all traditional medicines are for treatment of wounds or skin disorders, compared to only 1-3% of modern drugs. The use of such medicinal plant extracts for the treatment of skin disorders arguably has been based largely on historical/anecdotal evidence, since there has been relatively little data available in the scientific literature, particularly with regard to the efficacy of plant extracts in controlled clinical trials. In this article therefore, adverse and beneficial aspects of medicinal plants relating to skin and skin disorders have been reviewed, based on recently available information from the peer-reviewed scientific literature. Beneficial aspects of medicinal plants on skin include: healing of wounds and burn injuries (especially Aloe vera); antifungal, antiviral, antibacterial and acaricidal activity against skin infections such as acne, herpes and scabies (especially tea tree (Melaleuca alternifolia) oil); activity against inflammatory/immune disorders affecting skin (e.g. psoriasis); and anti-tumour promoting activity against skin cancer (identified using chemically-induced two-stage carcinogenesis in mice). Adverse effects of plants on skin reviewed include: irritant contact dermatitis caused mechanically (spines, irritant hairs) or by irritant chemicals in plant sap (especially members of the Ranunculaceae, Euphorbiaceae and Compositae plant families); phytophotodermatitis resulting from skin contamination by plants containing furocoumarins, and subsequent exposure to UV light (notably members of the Umbelliferae and Rutaceae plant families); and immediate (type I) or delayed hypersensitivity contact reactions mediated by the immune system in individuals sensitized to plants or plant products (e.g. peanut allergy, poison ivy (Toxicodendron) poisoning). PMID- 11482002 TI - Abraham Lincoln's blue pills. Did our 16th president suffer from mercury poisoning? AB - It is well known that Abraham Lincoln took a medicine called "blue mass" or "blue pill," commonly prescribed in the 19th century. What is now hardly known is that the main ingredient of blue mass was finely dispersed elemental mercury. As his friends understood, mercury was often prescribed for melancholy or "hypochondriasis," a condition Lincoln famously endured. Mercury in the form of the blue pill is a potential neurotoxin, which we have demonstrated by recreating and testing the recipe. We present the testimony of many of Lincoln's contemporaries to suggest that Lincoln suffered the neurobehavioural consequences of mercury intoxication but, perhaps crucial to history, before the main years of his presidency; he was astute enough to recognize the effects and stop the medication soon after his inauguration. PMID- 11482003 TI - Sunlight at Southall Green. Dr. Ingen Housz discovers photosynthesis. AB - In the following fictitious conversation, Dr. Jan Ingen Housz (1730-1799), the Dutch physician and natural philosopher, describes to William Temple Franklin (1760-1823), the grandson of Benjamin Franklin, how in 1779 he discovered the paramount role of sunlight in what we now call photosynthesis (Wiesner 1905; Van der Pas 1981; Reed 1949; Beale and Beale 1999). The two men, together with the English law reformer Samuel Romilly, were dinner guests of the First Marquis of Lansdowne at Lansdowne House on Wednesday 2 February 1791 (Bowood House Archives 1791). As far as possible we use their own recorded words and phrases, employing surviving manuscripts as a lexicon. Additional biographical and geographical details are provided in an Appendix, and all sources are listed in the References. PMID- 11482004 TI - Why is celiac disease so common in Ireland? AB - Celiac disease (gluten sensitive enteropathy) is a condition affecting the small bowel, characterized by permanent intolerance to dietary gluten, and giving rise to varying degrees of malabsorption and diarrhea. With the advent of sensitive screening tests, the condition is being increasingly diagnosed. Celiac disease is more common in the Irish and in those of Irish descent. Simoons (1978, 1981) hypothesized that the present-day prevalence of celiac disease across Europe is related to the interaction between genetic gradients, largely determined by the advance of agriculture, and historical patterns of cereal ingestion. This essay examines Simoons' hypothesis as it relates to Ireland, reviews the ethnic and genetic mix of those living on the island of Ireland and aspects of Irish dietary history, and considers how these factors may have combined to result in a high frequency of celiac disease. PMID- 11482005 TI - Plato's conception of the relations between moral philosophy and medicine. AB - The first part of this article construes crucial passages from the Platonic dialogues as evidence that Plato takes medicine as a model for moral philosophy and as one of the organizing principles for his writing of the dialogues. The second part reflects upon the significance of one of the most debatable implications of the model: the comparison between health and illness, on the one hand, and moral virtue and moral vice, on the other. It articulates 10 illuminating aspects of this comparison and two potentially serious objections to it. The third part of this article examines what the model implies about the roles of medicine and moral philosophy in the political community and about the natures of doctor-patient and philosopher-interlocutor relationships. It highlights Socrates' criticisms of a kind of politics that covers over moral causes of social disorders by means of Band-Aid legislation, and emphasizes that according to the model, both the doctor and the moral teacher confront agents who must cooperate in the process of reform and accept whatever responsibility they have for their conditions. PMID- 11482006 TI - William Crawford Gorgas (1854-1920). PMID- 11482007 TI - Albert Leon Charles Calmette (1863-1933) and the antituberculous BCG vaccination. AB - Albert Leon Charles Calmette was the first person to develop an anti-venom serum. His work revolutionized the treatment of snakebite in men and domestic animals. He is also well known for his development of the Bacillus Calmette-Guerin (BCG) vaccine for the prevention of tuberculosis. This article reviews the life experiences of this pioneer bioscientist, the current status of BCG in preventing tuberculosis, and the potential of BCG in immunotherapy for cancer. PMID- 11482008 TI - Human mobility and population health. New approaches in a globalizing world. AB - The globalization of economies in the last 25 years has greatly increased both the number of people on the move and the rapidity of their movement, and has brought attention to global disparities in health determinants and to the health of migrant populations themselves. Differences in epidemiological disease risk (prevalence gaps) may have negative, neutral, or positive health consequences for the migrant or receiving population. Population mobility represents a growing challenge to the development of public health programs and legislative policies to prevent the importation of disease, and to promote and protect the health of migrants and the local, receiving population. The inability to detect and contain imported disease threats at national borders requires a shift in immigration, quarantine, and public health approaches to health and mobile populations. A new paradigm is needed to facilitate the development of policies and programs to address the health consequences of population mobility. PMID- 11482009 TI - The problem of retaining clinical teachers in academic medicine. PMID- 11482010 TI - Notions of heredity in the correspondence of Edwin Grant Conklin. AB - This article examines five letters from the correspondence of American zoologist Edwin Grant Conklin that highlight his theories of genetic and social inheritance, in order to suggest that Conklin's eugenic beliefs--like those of many American authorities during this time--were complex and sometimes contradictory. The letters reveal the international prestige of American science after the two world wars and illuminate key moments in the emergence of the concepts of heredity and inheritance, within both the science of genetics and the social movement of eugenics. PMID- 11482011 TI - Evolutionary fallacies of Nazi psychiatry. Implications for current research. AB - Evolutionary theory has had a major impact on psychiatry since the middle of the 19th century. During the Nazi regime psychiatry supported compulsory sterilization and euthanasia of physically and mentally ill and subsequently the killing of "inferior" races by borrowing scientifically invalid conclusions from evolutionary biology. The present paper deals with some of the flaws and shortcomings of the scientific paradigms of evolutionary theory adopted by psychiatry during the Nazi regime and discusses possible implications for modern research in evolutionary psychology and psychiatry. PMID- 11482012 TI - Two against McCarthyism. Me and John P. Peters. PMID- 11482013 TI - [Fibrinolytic activity of duodenal mucosa in elderly and senile patients with the ulcer hemorrhage]. AB - The duodenal mucosa fibrinolytic activity was investigated in elderly and senile patients with chronic ulcer, complicated by hemorrhage. In noncomplicated duodenal ulcer the mucosal fibrinolytic activity of anterior intestinal wall prevailed such an activity of posterior wall. It preserved in the hemorrhage occurrence also. Maximal meaning of indices were noted on the second-fourth day after hemorrhage beginning in anterior duodenal wall and on the first day in posterior one. In term up to 8 days fibrinolytic activity had lowered, but in mucosa of anterior duodenal wall it exceeded by 44% the index in control group and of posterior wall--by 28%. Treating the patients it is necessary to take into account the results obtained, especially in presence of endoscopic signs of unstable hemostasis. PMID- 11482014 TI - [Application of endoscopic laser irradiation and antibacterial therapy in surgical treatment of duodenal ulcer disease]. AB - For duodenal ulcer disease there were performed an isolated selective proximal vagotomy--in 63 patients, vagotomy with various kinds of drainage operations--in 32, gastric resection according to Billroth-II--in 19. The expediency of application of endoscopic laser irradiation and antibacterial therapy in preoperative preparation complex for prophylaxis of early postoperative complications and the ulcer recurrences occurrence was noted. Among the patients operated on without the preoperative preparation conduction the inflammatory complications in early postoperative period occurred in 19.3% of them and the ulcer recurrence--in 11.5%; after preoperative preparation conduction--in 8.6 and 4.8% accordingly. PMID- 11482015 TI - [Diagnosis and treatment of perforative and "mirror-like" duodenal ulcer]. AB - The result of treatment of perforative duodenal ulcer (PDU) in 193 patients was analyzed. The PDU suturing was performed in 100 patients, radical organ preserving operation--in 93, with the PDU excision--in 80 of them. The "mirror like" ulcer of posterior wall was revealed in 12 (15%) patients using intraduodenal revision while doing the PDU excision. Selective proximal vagotomy with the PDU excision and processing of duodenal posterior wall is effective method of surgical treatment. PMID- 11482016 TI - [Desympathyzation of arteries in blood circulation disorders of upper extremities]. AB - Results of surgical treatment using the arteries desympathization in 47 patients with the upper extremities (UE) blood circulation disorder were studied. Artheriolysis was performed in 21, periarterial sympathectomy--in 26 patients. For obliterating disease 19 patients were operated, for Raynaud syndrome--9, for the traumatized vessel diseases--8, for the thoracic outlet compression syndrome- 11. Desympathization of arteries secures significant improvement of blood circulation in distal parts of UE in terms up to 1 mo in 95.7% of the patients operated on. PMID- 11482017 TI - [Application of clexane for prophylaxis of thromboembolic complications in surgery of morbid obesity]. AB - The results of comparative clinical application of low-molecular heparin Clexan vs heparin for the thromboembolic complications prophylaxis in surgical treatment of morbid obesity were summarized. PMID- 11482018 TI - [Expediency of the liposuction in surgery of abdominal surgery, abdominal diseases and small organs of the pelvis]. AB - The experience of performance of dermatolipectomy in 124 patients while conducting operative intervention for abdominal hernia and for the abdominal cavity and the small pelvis organs diseases was summarized. The risk degree of the simultant operation performance was estimated while conducting the proper preoperative preparation and the surgical correction volume was planned beforehand. The simultant operation performance did not influence the stationary treatment duration significantly, it also permitted to reduce the duration of temporary disability to shorten significantly the treatment expenses, to stop the stress influence on patient's psychological state caused be the reoperation conduction necessity. Low-molecular heparin fraxiparin (produced by Sanofi firm) was applied for the thromboembolic complications prophylaxis under control of the blood coagulation time and of the coagulogramm indices. PMID- 11482019 TI - [New approaches to the treatment and prophylaxis of the cicatrix formation]. AB - Preparation Contractubex (gel for external application in tubes of 20 g each), manufactured by pharmaceutical company Merz GmbH (Frankfurt-on-Main, Germany), was used during the treatment of 30 patients with hypertrophic cicatrix and contracture. PMID- 11482020 TI - [Efficacy of vascular therapy in complex of treatment of purulent necrotic foot ulcers in patients with diabetes mellitus]. AB - The result of treatment of purulent-necrotic affection of foot (PNAF) in 151 patients with diabetes mellitus (DM) was estimated. The conduction of preoperative medicinal therapy is mandatory for the general status of patient stabilization only. After prescribing of heparin, no-spa, papaverin, rheopolyglucinum, solcoseryl, aspirin the foot tissues blood supply is changing nonvitally, the foot tissues blood supply while the alprostan intravenous infusion improves significantly. The alprostan prescription during performance of operative intervention on foot in patients with PNAF, caused by DM, and in the collateral blood flow decompensation of the leg and foot arteries is mandatory. PMID- 11482021 TI - [Application of endolymphatic therapy using catheterization of femoral lymphatic vessels in complex of treatment of oncological diseases]. AB - Endolymphatic way of the medicinal preparations administration promotes their more effective influence on pathological focus. The experience of conduction of more then 15,000 of the lymphatic vessels catheterization manipulations was summarized, permitting to perform the treatment of patients successfully. PMID- 11482022 TI - [Clinico-statistical substantiation of the cryosurgery application in treatment of glial tumors]. AB - The results of application of cryosurgical technology in treatment of 146 patients with glial tumor of various grade of anaplasia were analyzed. The prognostic significance of principal clinico-laboratory and instrumental indices was studied. The disease prognosis is determined by sum of diagnostical coefficients, prognostically favourable and unfavorable factors present. The higher efficacy of application of cryosurgical method in comparison with generally accepted method of surgical treatment was established. PMID- 11482023 TI - [Morphological substantiation of choice of the laser processing of residual cavity wall after hydatidectomy for pulmonary hydatid cyst]. AB - Morphological investigation of the pulmonary echinococcal cyst after applying the laser irradiation influence on it was done. The optimal way for performance of an adequate scolexo- and bactericidal treatment of fibrous capsule after the pulmonary hydatidectomy conduction is application of defocused to 5-10 mm irradiation of the CO2 and the Nd AIG-laser with 150-250 Wt/cm2 power density. In the cyst suppuration it is preferable to use Nd AIG-laser with increase of the exposition time in 3-4 times and of the energy absorbance up to 400-800 J/cm2. Local coagulation using irradiation of the Nd AIG-laser by the contact method with the 20 Wt power output (250-400 Wt/cm2 power density) is applied if bronchi, which are confided into residual cavity, have diameter up to 3 mm. PMID- 11482024 TI - [Diagnosis, strategies of treatment of an acute hematogenic osteomyelitis in children]. AB - There were examined 275 children with an acute hematogenic osteomyelitis (AHO). The inflammatory focus puncture has decisive diagnostic significance. The local pH determination was applied as a method of the AHO express diagnosis. Early diagnosis and conduction of complex treatment during three days from the disease beginning had provided the patient's recovery. PMID- 11482025 TI - [Operative treatment of fecal incontinence in coccyx agenesis in children after surgical correction of Hirschsprung's disease]. AB - The coccygeal agenesis is one of causes of the external anal sphincter functional insufficiency after performing operations for Hirschsprung's disease in children. The frequency of its diagnosis had constituted 6.2%, in 14.6% it was the causative factor of fecal incontinence after the operation. In coccygeal agenesis the normal anatomy of the pelvic floor muscles become disordered. It is convenient to perform the anomaly correction during the radical operation conduction for the main disease. PMID- 11482026 TI - [Microbiological aspects of application of fluoroquinolones in surgery]. AB - Antimicrobic activity of fluorochinolons with regard to pathogenic organisms of purulent-septic surgical infection was studied. In comparison with cephalosporins of the third generation, higher activity of fluorochinolons is noted with regard to majority aerobic microorganisms, and to pseudomonads, in particular. Optimal pharmacokinetics properties, good compliance, wide antimicrobic spectrum permits to use fluorochinolons in conduction of empirical antibacterial therapy for the infection occurrence prophylaxis in surgical practice, and for the selective intestinal decontamination, in particular. PMID- 11482027 TI - [Anesthesiological support of operative treatment in patients with infectious toxic shock]. AB - The state of the vitally significant functions in patients with sepsis and infective-toxic shock (ITS), to whom operative intervention was done, was studied. The expediency of application in postoperative period of hypertonic and hypertonic hyperoncotic solutions in combination adrenomimetic preparations was proved. The advantages of multicomponent anesthesia, based on oxybate and ketamine in comparison with neuroleptanalgesia and ataralgesia were substantiated. The method of the anesthesiological securing was proposed, application of which have permitted to raise essentially the safety of the operative intervention in patients with ITS. PMID- 11482028 TI - [Choice of surgical strategies in the treatment of multiple postnecrotic pancreatic pseudocysts]. AB - The results of treatment of patients with multiple pancreatic pseudocysts are presented. The surgical tactics choice, depending on size, localization of pseudocysts, the state of their wall and connection with pancreatic duct, was substantiated. PMID- 11482029 TI - [Microbiological monitoring of the microbic strain stability to antibacterial preparations in surgical in-patient clinics]. AB - While performing of microbiological monitoring, conducted in surgical stationaries of Dnepropetrovsk, of 775 cultures, isolated during one year from patients and outer space, the S. aureus cultures were revealed, resistant to methicillin were 23.6%, to gentamycin 12.3% culture of all Gram-negative bacteria. Most effective preparation, which is recommended for the empirical antibacterial therapy conduction, is meronem, to whom the sensitivity of 87.5% for Gram-positive and 92.1% for Gram-negative cultures of conventionally pathogenic microorganism were established. PMID- 11482030 TI - [Choice of systemic treatment of the invasive mammary gland cancer]. PMID- 11482031 TI - [The ureter's duplication megaureter of the right kidney, complicated by purulent pyelonephritis simulating infected pancreatic cyst]. PMID- 11482032 TI - [Observation of successful surgical treatment of seven tumors in a woman patient with genetically complicated heredity]. PMID- 11482033 TI - [Application of mini-invasive interventions in combination with video thoracoscopy for diseases of the thoracic cavity organs]. PMID- 11482034 TI - [Revisional total endoprosthesis of a knee joint]. PMID- 11482035 TI - [Observation of catamenial hemopneumothorax]. PMID- 11482036 TI - [Observation of anomalous location of hepatic arteries]. PMID- 11482037 TI - [Typological peculiarities of giant gastric ulcer]. AB - Of 1441 patients operated on, the giant gastric ulcer was revealed in 16.6%. It is characterized by polymorphism: several ulcers were revealed in 7.6%, coexistent gastric and duodenal ulcer--in 19.8% and, in contradistinction to genuine gastric ulcer (I type--according to Johnson), by high frequency of such complications as hemorrhage (in 44.1% of observations), penetration (in 74.1%), malignancy (in 14.3%). PMID- 11482038 TI - Treatment of chronic hepatitis C in refractory patients. What is the answer? PMID- 11482039 TI - Long-term response to interferon plus ribavirin in patients with chronic hepatitis C refractory to interferon. AB - OBJECTIVE: A sustained response (SR) to interferon (IFN) is only observed in 15 20% of patients with chronic hepatitis C (CHC). The aim of this study was to determine the long-term effectiveness and safety of the treatment with IFN plus ribavirin (RIB) over two years in CHC patients without SR to IFN. DESIGN: A prospective and open longitudinal follow-up study was conducted over 3 years. PATIENTS AND METHODS: A total of 77 CHC patients were included: 63 non-responders (NR) and 14 relapsers (R) to IFN. Patients were treated with IFN (3 MU s.c. three times a week) and RIB (1,000-1,200 mg p.o. daily) for 12 months. Treatment tolerance and viral response (HCV-RNA in serum < 1,000 copies/ml) were assessed after 1, 3, 6 and 12 months of treatment. SR and relapsing rates were subsequently evaluated 6, 12 and 24 months after the end of the treatment, together with those variables capable of predicting SR. RESULTS: At the end of the treatment, 19/77 patients responded (24.7%), 9/63 (14.3%) were non-responders and 10/14 (71.4%) relapsers, and these same patients exhibited SR after 6 months. The SR rate two years after treatment was 22.1% [8/63 (12.7%) NR and 9/14 (64.3%) R]. The relapse rate after 6 months and two years was respectively 0 and 10.5% (2/77). Independent variables capable of predicting SR were negative viremia conversion within the first month of treatment, maintenance of such negative viremia after 6 months, and R status to IFN. Side effects were recorded in 90.9% of cases (70/77), the most frequent being pseudoinfluenza syndrome. Treatment had to be discontinued in 33.8% of patients (26/77). CONCLUSIONS: Combined IFN-RIB therapy for 12 months in CHC patients without SR to IFN obtains a long-term SR of 22.1%, this rate being higher in relapsers to prior IFN therapy (64.3% in R versus 12.7% in NR). PMID- 11482040 TI - Anal endosonography for the study of anal canal anatomy: normal images and sonographic variants. AB - OBJECTIVE: To present normal images and sonographic variants of the anal canal to be used as reference for the study of sphincter and anal canal abnormalities. MATERIAL AND METHODS: Sixty subjects without known anal canal disease were studied by means of anal endosonography. Subject were divided according to age in two groups (up to 50 years and more than 50 years). All of them underwent an outpatient study with B&K medical ultrasound 2,003 scanner and 1,850 multifrequency transducer. RESULTS: Four layers can be sonographically identified in the anal canal: an inner hyperechoic layer which is the submucosa, a second hypoechoic layer which is the internal sphincter, a third one which is a longitudinal muscle and the outer hyperechoic layer which is the external sphincter and the only to be found in the low anal canal. In people older than 50 years, both sphincters were significantly thicker (0.3-0.5 mm). At the high anal canal 40% of women presented an anterior gap in the external anal sphincter. CONCLUSIONS: Anal endosonography allows an easy division in high-, mid-, and low anal canal. In some women there is a gap at the mid-high anal canal that must be taken into account in order to avoid diagnostic errors. An internal sphincter thickness greater than 3.5 mm should be considered abnormal at any sex or age. PMID- 11482041 TI - Helicobacter pylori eradication versus one-year maintenance therapy: effect on relapse and gastritis outcome. AB - OBJECTIVE: The aim of this study was to determine ulcer healing and H. pylori eradication rates obtained with triple therapy (omeprazole, amoxicillin and clarithromycin). Ulcer relapsing rate one year after eradication was also assessed. Maintenance therapy with placebo was compared with ranitidine therapy and the effect of eradication on histological variables of the gastric mucosa was studied. METHODS: A prospective, double-blind parallel study was performed in 85 patients endoscopically diagnosed of duodenal ulcer H. pylori positive. Patients were randomized to a 7-days triple therapy (group A) or omeprazole plus antibiotic placebo (group B). All patients were treated only with omeprazole for the next three weeks. Patients with ulcer healing after treatment were entered in a one-year follow up phase with ranitidine placebo (group A) or ranitidine (group B). Endoscopy and biopsies were performed at baseline, after treatment (5 weeks) and after 12 months of follow-up or when relapsing symptoms appeared. RESULTS: Healing rate was 90.2% in group A and 85.7% in group B. Eradication rate was 78% in group A and 0% in group B. Out of 37 healed patients in group A, eradication was achieved in 29 and only one relapse was found (3.4%). Three out of eight patients with healing but without eradication relapsed at 12 months (35%) (p < 0.05). Histopathological results showed statistically significant differences (p < 0.05) between eradicated and non eradicated patients in terms of severity of inflammation and intestinal metaplasia, but not in terms of atrophy. CONCLUSIONS: H. pylori eradication is useful to prevent ulcer relapse and to improve gastric mucosa status. PMID- 11482042 TI - Assessment of hospital costs related to the diagnosis and treatment of upper gastrointestinal haemorrhages in patients consuming non steroid anti-inflammatory drugs. AB - BACKGROUND: Non-steroid anti-inflammatory drugs (NSAIDs) can produce haemorrhages of the digestive tract, whose treatment result in significant hospital costs. OBJECTIVE: The aim of this analysis has been to estimate hospital costs related to the treatment of digestive haemorrhages potentially caused by the intake of NSAIDs. MATERIAL AND METHODS: The study was conducted by reviewing the clinical history of all patients admitted in two tertiary Spanish hospitals during 1998 due to digestive haemorrhage following NSAID treatment. After the identification of cases, all resources consumed during their hospitalisation (concomitant medication, complementary examinations and tests, surgery, blood products consumption, inpatient consultations and length of stay in the hospital) until the complete resolution of each case, were recorded. RESULTS: Thirty-six percent of patients admitted due to digestive haemorrhage had taken some NSAID the same day of their hospitalisation (85.4%) or in previous days. The cost related to the treatment of these patients amounted to some 71 million pesetas for both hospitals, with a cost/patient of 434,407 pesetas. CONCLUSIONS: Given the high consumption of NSAIDs in our normal clinical setting, costs related to the diagnosis and treatment of digestive haemorrhages in hospitals are a highly significant burden for the National Health System. PMID- 11482043 TI - Colorectal cancer experimental models. PMID- 11482044 TI - [Acute perforated appendicitis. Manifestation of incarcerated inguinal hernia]. PMID- 11482045 TI - [Gastric perforation as first manifestation of extracutaneous dissemination of mycosis fungoides]. PMID- 11482046 TI - [Electrolyte levels and polyethylene glycol administration]. PMID- 11482047 TI - [Salmonella typhimurium gastroenteritis and erythema nodosum]. PMID- 11482048 TI - [Colonic bezoar]. PMID- 11482049 TI - The glycemic load. PMID- 11482050 TI - Spirituality and spiritual care provided by parish nurses. AB - The high level of religious participation in the United States provides a venue for parish nursing, a holistic nursing specialty that emphasizes the relationship between spirituality and health. This descriptive study measured two aspects of spirituality (spiritual perspective and spiritual well-being) in a national sample of parish nurses and described variables related to their practice. Furthermore, it qualitatively examined the provision of spiritual care to clients in this parish nurse sample. Parish nurses scored high in spiritual perspective and spiritual well-being and reported an emphasis on health promotion and education in their activities. Three views of spiritual interventions (ideal, general, and specific) were reported. Types of spiritual interventions typically fell into one of four categories: religious, interactional, relational, and professional. PMID- 11482051 TI - Violence, risk, and survival strategies of street prostitution. AB - The purpose of this qualitative study is to explore the risks and responses of street prostitutes to customer-related violence. Thirteen female street prostitutes, aged 18 to 40, were recruited from a moderate-size, midwestern city in the United States. Eight were non-Hispanic White and five were African American. Respondents participated in individual interviews to identify perceptions of work-related violence, personal risk, and self-protective survival strategies. The constant comparative method was used to analyze data. PMID- 11482052 TI - The interaction of age and cognitive representations in predicting blood pressure. AB - The purpose of this study was to investigate the interaction of age and cognitive representations of hypertension in predicting blood pressure. A model of illness self-regulation was used as the theoretical framework for the study. Secondary analysis of data collected from 224 hypertensive adult outpatients was conducted to assess five constructs of illness. ANOVA results indicated that older adults reported fewer hypertension-related symptoms than younger- and middle-aged adults, and that younger adults reported fewer health behaviors to control hypertension than did middle-aged and older adults. Results of hierarchical multiple regression models indicated that after adjusting for effects of demographic factors, no single cognitive representation construct was a significant predictor of blood pressure. The interactions of age and three cognitive representation constructs were, however, significant predictors of blood pressure. For younger adults, increases in the perceived Consequences, Control, and Timeline for hypertension were predictive of decreases in hypertension, although for older adults increased perceptions of control were associated with increased blood pressure. PMID- 11482053 TI - Social cognitive factors predicting the health of elders. AB - The social cognitive model of health is primarily concerned with influences on a person's decisions to perform health behaviors. In this study, the relationships among social cognitive factors reflecting enabling skills (learned resourcefulness), internal motivation for health (health self-determinism), and help responses (coping responses) and a measure of physical and psychosocial health were examined in 137 chronically ill elders. Gender, race, and number of chronic conditions were predictors of learned resourcefulness. Learned resourcefulness was associated with the use of informal help, whereas health self determinism was related to self-help and formal help. None of the help responses were significant predictors of health. Model testing revealed that greater resourcefulness or skill in coping with stressful situations was the most important predictor of health. Strategies are recommended to help elders achieve optimal health. PMID- 11482054 TI - Using collaborative research to facilitate student learning. AB - Developing research partnerships between academia and the service sector is an innovative way to meet the demand for high-quality, cost-effective, and clinically oriented research. Undergraduate student participation in clinical research is an educational strategy to facilitate positive mindsets toward research. This article outlines the methodological steps in recruiting and training undergraduate students for clinical research teams to benefit nurse educators, nurse researchers, students, and institutional partners. Student volunteers collected data for a study examining patient satisfaction with pain management practices. The research proposal was used to demonstrate principles of the research process and to familiarize the students with the study. A detailed study protocol guided the entire team through the project. Student sensitivity to pain assessment and management was enhanced. Learning the research process and the students' appreciation for the rigors of research were reinforced using this experiential model. Student evaluation of the research experience is presented. PMID- 11482055 TI - A metaphor for HIV-positive Mexican and Puerto Rican women. AB - Human Immunodeficiency Virus (HIV) represents an overwhelming disruption for Hispanic women. This Grounded Theory (GT) (Glaser & Strauss, 1967) study sought to explore how culture guided health practices and to identify health and folkcare activities used by HIV+ Hispanic women. La Protectora (The Protectress), a theoretical framework, emerged to describe the process by which cultural beliefs guided Hispanic women's health practices and how they dealt with life after their affirmation of HIV status. Five main categories emerged to describe the process. Knowledge about how culture influences HIV+ Hispanic women obtain and/or make decisions about health care will contribute to a foundation for theory development and subsequent theory testing. PMID- 11482056 TI - Perceptions of role functions of psychiatric nurse specialists. AB - Specialized and advanced psychiatric nursing practice is an innovative concept in the health care service in Hong Kong. A clear definition of the role and practice of psychiatric clinical nurse specialists (CNS) is important for the development of expert psychiatric nursing practice but is still in a formative stage. A qualitative exploratory study was conducted to identify the psychiatric CNS's perceptions of their role and to compare their perceptions with those of their clinical psychiatric nurse colleagues. The main themes emerging from interviews, observations, and personal diary data were compared and condensed. Eight CNSs and 24 clinical nurse colleagues from acute care and community psychiatric nursing units voluntarily participated in the study. A four-component framework, including clinical practice, organization, education, and professional role, was adopted from nursing literature and used for categorization. The findings showed similarities of role perceptions in the clinical practice, organization, and education components between the CNSs and their nursing colleagues. Differences in role perceptions, concerns about inadequate knowledge and autonomy, and limitations in professional role are discussed. PMID- 11482057 TI - The influence of the analogue 4-10 corticotropin derivative on the general structure of the rabbits behaviour in stress. PMID- 11482058 TI - Long-term survival among patients with chronic lymphocytic leukemia (CLL) treated in Department of Hematology, Medical University of Lublin. PMID- 11482059 TI - Breast cancer--the role of free radicals and antioxidants in pathogenesis of benign dysplasia and breast carcinoma. PMID- 11482060 TI - Ultrasonographic estimation of spleen size in children affected with vitiligo. PMID- 11482061 TI - Analysis of causes and effects of children's injuries in a rural area. PMID- 11482062 TI - The drug susceptibility of Staphylococcus aureus strains isolated from nose and pharynx. PMID- 11482063 TI - Experimental model of broncho-pleural fistula. PMID- 11482064 TI - Assessment of respiratory system efficiency during experimental thoracotomy with lung tissue resection in sheep based on arterial gas blood analysis. PMID- 11482065 TI - Lung function in chronic uremia. PMID- 11482066 TI - Arm lymphoedema after surgical treatment for the cancer of the breast. PMID- 11482067 TI - Complications of axillary node dissection for breast carcinoma as perceived by patients. PMID- 11482068 TI - Oxidative stress in respiratory tract diseases in babies. PMID- 11482069 TI - An assessment of the osteoporosis changes in rat mandible using the scanning electron microscope (SEM). PMID- 11482070 TI - The influence of TP-1 on the behaviour of rabbits in spontaneous conditions and after stimulation of ventromedial hypothalamic nucleus (Vmh). PMID- 11482071 TI - The influence of solcoseryl and vincristine on the mechanical strength of the femur shaft in young rats. PMID- 11482072 TI - Effects of experimental diabetes on changes in the eye ball external muscles in rabbit. PMID- 11482073 TI - Treatment needs of stomatognatic system of 12-year-old children from the Lublin region. PMID- 11482074 TI - A delayed replantation of permanent upper incisors. PMID- 11482075 TI - The influence of normobaric hyperoxia on antioxidant enzyme activities and peroxidation products levels in rat brain. PMID- 11482076 TI - The influence of tuftsine analog TP-1 on the behaviour of rabbits. PMID- 11482077 TI - The effect of cladribine on some parameters of blood and cerebrospinal fluid in patients with relapsing-remitting multiple sclerosis (RR-MS). PMID- 11482078 TI - First neurological symptoms of multiple sclerosis. PMID- 11482079 TI - The levels of C3 and C4 components of the complement in the sera of relapsing remitting multiple sclerosis patients (RR-MS) treated by 2-CDA (cladribine). PMID- 11482080 TI - The problem of drug addiction among secondary school students in Lublin. Young people's knowledge about the effects of taking drugs. PMID- 11482081 TI - Diagnostic difficulties in cystic renal tumors. PMID- 11482082 TI - Examining of changes in the structure of rat bone maxilla and mandible treated with Hydrocortisonum hemisuccinatum. PMID- 11482083 TI - The influence of apomorphine on the cardiovascular reactions induced by stimulation of the vagus nerve in the rabbit. PMID- 11482084 TI - Plasma level of IL-8 in patients with psoriasis and its correlation with psoriasis area and severity index and the clinical type of the disease. PMID- 11482085 TI - Comparison of serum lipid in girls affected with vitiligo and control group. PMID- 11482086 TI - Features of proteinuria in rat kidney in experimental nephrotic syndrome. PMID- 11482087 TI - The histological and ultrastructural picture of the paraepidermoidal epithelium subjected to low temperatures. I. PMID- 11482088 TI - The histological and ultrastructural picture of the paraepidermoidal epithelium subjected to low temperatures. II. PMID- 11482089 TI - Study on activity of interleukin-6, C-reactive protein and alpha-2 macroglobulin in psoriatic patients during the acute clinical stage and remission. PMID- 11482090 TI - The histological and ultrastructural picture of the paraepidermoidal epithelium subjected to low temperatures. III. PMID- 11482091 TI - Impacted maxillary canines--evaluation of reasons of occurrence and complications on the basis of own observations. PMID- 11482092 TI - Diagnostic algorithm in high (cranial) maxillofacial fractures. PMID- 11482093 TI - Epidemiological situation of tuberculosis in Poland at the end of the 20th century. PMID- 11482094 TI - The phenomenon of "nicotinism" among teenagers living in state orphanages in the city of Lublin. PMID- 11482095 TI - Problems of multiple body injuries in children. PMID- 11482096 TI - Biomechanical prenatal factors for the development of congenital hip dysplasia. PMID- 11482097 TI - The concept of holistic education as the alternative to technologisation of medicine. PMID- 11482098 TI - Evaluation of abilities to recognise child maltreatment syndrome among doctors. PMID- 11482099 TI - Knowledge of burns management among citizens of Lublin. PMID- 11482100 TI - Action of 1,2,4-triazole and 1,2,4-triazine derivatives on cells of green monkey kidney in in vitro culture. PMID- 11482101 TI - Family--opinions and expectations of alcoholics from AA (Anonymous Alcoholics) groups. PMID- 11482102 TI - Blood serum TNF-alpha concentration in patients with ischaemic and haemorrhagic brain stroke. PMID- 11482103 TI - Serotonin in pregnancies complicated by intrahepatic cholestasis. PMID- 11482104 TI - Virtual computed tomography in the evaluation of the vertebral canal. PMID- 11482105 TI - Injuries of the middle ear with a welding spark. PMID- 11482106 TI - Effects of cisplatine on the level of phospholipids in brain tissue of the rat. PMID- 11482107 TI - Injuries sustained by children in road accidents in the Lublin province as a cause of hospitalisation. PMID- 11482108 TI - Knowledge of first-aid rules among students. PMID- 11482109 TI - Natural and civilization emergencies in Poland--general feelings of security. PMID- 11482110 TI - Evaluation of immunoglobulins, proinflammatory cytokines and lipid concentrations in patients with diabetic nephropathy treated with continual ambulatory peritoneal dialysis (CAPD). PMID- 11482111 TI - Cutaneous nerves of the brachial plexus in postnatal life in man. PMID- 11482112 TI - The senses of smell and taste in anorexia. PMID- 11482113 TI - Voice disorders in children with gastroesophageal reflux disease. PMID- 11482114 TI - The sense of smell and the behaviour of nasal mucociliary transport in allergic children. PMID- 11482115 TI - Disturbances of voice in children with atopic diseases. PMID- 11482116 TI - Ambulatory and community-based services. AB - The shift in the site of service delivery from inpatient and institutional to ambulatory and community settings has been prompted by concerns over cost and the prospect for improving the quality of life. In response to these concerns, Medicare has implemented several demonstrations that emphasize ambulatory and community-based services. In this issue, articles are presented on four demonstrations, which focus on the extent to which coordinated care models reduce health care costs, and the cost effectiveness and beneficiary outcomes of disease specific programs. Two articles are included on home health. One examines home health care in relation to the other Medicare post-acute benefits, and the other focuses on the use of home health care in the treatment of end stage renal disease (ESRD). Finally, two articles report on Section 1915c Medicaid home and community-based waiver programs. PMID- 11482117 TI - Impact of the BBA on post-acute utilization. AB - In this article, the author summarizes recent changes in Medicare post-acute payment policies, discusses the implications of certain design and implementation issues, and analyzes whether different types of patients are using skilled nursing facilities (SNFs), home health agencies (HHAs), and rehabilitation hospitals and units. If similar populations are treated by these three types of providers, service patterns may be affected by the financial incentives in the new, more restrictive payment policies. The author describes new post-acute care (PAC) payment policies, service patterns prior to the Balanced Budget Act of 1997 (BBA), differences in the populations using these providers, and possible effects of the new payment systems on site-of-care decisions. PMID- 11482118 TI - Use of home health care by ESRD and Medicare beneficiaries. AB - The use of home health care (HHC) services among Medicare end stage renal disease (ESRD) enrollees remains an under-studied area. In this article, the authors report sociodemographic characteristics and patterns of HHC utilization by Medicare-covered ESRD patients. The authors found that those who were female, age 85 or over, diabetic, and residing in the New England or West South Central census divisions were more likely to use HHC services and were also more intensive users. Analysis of use patterns in such high-risk populations is necessary to ensure that health policy changes do not have unintended consequences for vulnerable patients. PMID- 11482119 TI - Trends and issues in the Medicaid 1915(c) waiver program. AB - Over the past 15 years, Medicaid 1915(c) home and community-based waivers have made a substantial contribution to States' efforts to transform their long-term care (LTC) systems from largely institutional to community-based systems. By 1997, every State had implemented a waiver program for at least some subgroups of individuals with disabilities, and expenditures increased from $3.8 million in 1982 to more than $8.1 billion in 1997. Emerging, as well as long-standing, policy issues related to the waiver program include concerns with access, variation in availability by disability group, State decisions related to the provision of community-based LTC, and evidence on effectiveness. PMID- 11482120 TI - Home-care use and expenditures among Medicaid beneficiaries with AIDS. AB - This article compares the use and cost of home-care services among traditional Medicaid recipients with acquired immunodeficiency syndrome (AIDS) and among participants in a statewide Human Immunodeficiency Virus (HIV)/AIDS-specific home and community-based Medicaid waiver program in New Jersey, using Medicaid claims and AIDS surveillance data. Waiver program participation appears to mitigate racial and risk group differences in the probability of home-care use. However, the program's successes are confined to its enrollees of which subgroups of the AIDS population are underrepresented. Our findings suggest the need to expand access to home-care programs to racial minorities and injection drug users (IDUs) with HIV/AIDS. PMID- 11482121 TI - Cost of smoking to the Medicare program, 1993. AB - Medicare expenditures attributable to smoking in 1993 were estimated using a multivariate model that related expenditures to smoking history, health status, and the propensity to have had a smoking-related disease, controlling for sociodemographics, economic variables, and other risk factors. Smoking attributable Medicare expenditures are presented separately for each State and by type of expenditure. Nationally, smoking accounted for 9.4 percent of Medicare expenditures--$14.2 billion, with considerable variation among States. Smoking accounted for 11.4 percent of Medicare expenditures for hospital care, 11.3 percent of nursing home care, 5.9 percent of home health care, and 5.6 percent of ambulatory care. PMID- 11482122 TI - Selection experiences in Medicare HMOs: pre-enrollment expenditures. AB - Using 1993 and 1994 data, the authors examine whether beneficiaries who enroll in a Medicare health maintenance organization (HMO), including those enrolling for only a short period of time, have lower expenditures than continuous fee-for service (FFS) beneficiaries the year prior to enrollment. We also test whether biased selection varies by the level of HMO market penetration and the rate of market-share growth. We find favorable selection associated with enrollment into Medicare HMOs, which declines as market share increases but does not disappear. Among short-term enrollees, we find unfavorable selection, however, selection bias was not sensitive to market characteristics. PMID- 11482123 TI - Cost and outcomes of Medicare reimbursement for HMO preventive services. AB - Medicare beneficiaries enrolled in a health maintenance organization (HMO) were randomized to a preventive services benefit package for 2 years or to usual care. At 24- and 48-month follow-ups, the treatment group had completed more advance directives, participated in more exercise, and consumed less dietary fat than the control group. Unexpectedly, more deaths occurred in the treatment group. Surviving treatment-group enrollees reported higher satisfaction with health, less decline in self-rated health status, and fewer depressive symptoms than surviving control participants. Despite these changes, the intervention did not yield lower cost per quality-adjusted life year in this historically prevention oriented HMO. PMID- 11482124 TI - Effects of the Medicare Alzheimer's disease demonstration on Medicare expenditures. AB - Applicants were randomized either into a group with a limited Medicare community care service benefit and case management or into a control group receiving their regular medical care. Analyses assess whether or not community care management affected health care use. A tendency toward reduced expenditures was observed for the treatment group, combining all demonstration sites, and when observing each separately. These differences were or approached statistical significance in two sites for Medicare Part A and Parts A and B expenditures averaged over 3 years. Expenditure reductions approached budget neutrality with program costs in two sites. PMID- 11482125 TI - Long-term care eligibility criteria for people with Alzheimer's disease. AB - Long-term care (LTC) eligibility criteria are applied to a sample of 8,437 people with dementia enrolled in the Medicare Alzheimer's Disease Demonstration. The authors find that mental-status-test cutoff points substantially affect the pool of potential beneficiaries. Functional criteria alone leave out people with relatively severe dementia and with behavioral problems. It is therefore important to consider both behavioral and mental-status-test criteria in establishing eligibility for community-based services for people with dementia. PMID- 11482126 TI - S/HMO versus TEFRA HMO enrollees: analysis of expenditures. AB - This study compares expenditures on health care services for enrollees in a social health maintenance organization (S/HMO) and a Tax Equity and Fiscal Responsibility Act of 1982 (TEFRA)-risk Medicare health maintenance organization (HMO). In addition to the traditional Medicare services covered by the TEFRA HMO, the S/HMO provided a long-term care (LTC) benefit and case management services for chronic illness. There do not appear to be any overall savings associated with S/HMO membership, including any savings from substitution of S/HMO-specific services for other, traditional services covered by both the S/HMO and the TEFRA HMO. PMID- 11482127 TI - Case management for high-cost Medicare beneficiaries. AB - We estimated the effects of three Health Care Financing Administration (HCFA) funded case management demonstrations for high-cost Medicare beneficiaries in the fee-for-service (FFS) sector. Participating beneficiaries were randomly assigned to receive case management plus regular Medicare benefits or regular benefits only. None of the demonstrations improved self-care or health or reduced Medicare spending. Despite the lack of effects of these interventions, case management might be cost-effective if it includes greater involvement of physicians, is more well-defined and goal-oriented, and incorporates financial incentives to generate savings in Medicare costs. Models incorporating these changes should be investigated before abandoning Medicare case management interventions. PMID- 11482128 TI - The effects of arecoline on the release of cytokines using cultured peripheral blood mononuclear cells from patients with oral mucous diseases. AB - In Taiwan there is a significant correlation between oral precancer diseases and oral cancer associated with the betel quid chewing habit. The carcinogenic components of betel quid are arecoline, arecaidine and safrole. However, it is unknown whether these substances influence the immune functions. This study investigated the effects of betel quid on the immune system using cultured peripheral blood mononuclear cells from patients with oral mucous diseases. In our experiment, mononuclear cells from 10 normal persons, 12 patients with precancer lesions, and 16 patients with squamous cell carcinoma were separated from blood samples and cultured. After stimulation by arecoline, the amounts of IL-2, TNF-alpha, TGF-beta and IFN-gamma secreted by mononuclear cells were measured using the ELISA method. The results showed that IL-2, TNF-alpha, and TGF beta were significantly lower in mononuclear cells of normal persons as stimulated by arecoline. The TGF-beta amount in cells from oral submucous fibrosis patients with betel quid chewing habit (OSF-B) was lower than normal persons or patients who had long term betel quid chewing habit but were without oral mucosal diseases (N-B), and was also lower than the squamous cell carcinoma with betel quid chewing group (SCC-B). TNF-alpha was significantly lower in the squamous cell carcinoma with long term betel quid chewing group (SCC-B) than in normal persons. TNF-alpha was significantly higher in the squamous cell carcinoma without betel quid chewing group (SCC-N) than in normal persons and SCC-B groups. In addition, IFN-gamma was significantly lower in patients who had long term betel quid chewing but were without oral mucous lesions than the normal person and the OSF group. The results proved that betel quid influences cytokines production by mononuclear cells. PMID- 11482129 TI - HLA-DQA1 genotyping in patients with rheumatoid arthritis in Taiwan. AB - To investigate the role of HLA-DQA1 genotypes and their interaction with HLA-DRB1 in the pathogenesis of rheumatoid arthritis (RA) in Taiwan, HLA-DQA1 was determined in 71 patients with RA and 108 healthy controls by SSP-PCR method. HLA DRB1 and HLA-DQA1 were simultaneously detected in 55 RA patients and 101 healthy controls. PCR/SSOP method was used to determine the HLA-DRB1 genotypes, and the subtypes of HLA-DR4 were determined by cloning and sequencing. The phenotypic frequency of HLA-DQA1*0301 was significantly lower in RA than in controls, and, in contrast, the HLA-DQA1*0302 and DQA1*0303 were significantly higher in RA than in controls. The associations of DQA1*0301, *0302, and *0303 with RA were independent of DR4 and DRB1*0405. Moreover, the interactions between HLA-DR4 and HLA-DQA1*0302 or DQA1*0303 could enhance the development of RA. We also found that the prevalence of bone erosion and seropositivity of rheumatoid factor (RF) were significantly higher in HLA-DQA1*0303 positive RA patients than in healthy controls. HLA-DQA1*0302 and DQA1*0303 are the risk factors for susceptibility to RA, while HLA-DQA1*0301 is a protective factor. A synergistic effect for the susceptibility to RA can be found between HLA-DR4 and HLA-DQA1*0302 or DQA1*0303. We also found that the HLA-DQA1*0303 was related to bone erosion and seropositivity of RF in RA patients. PMID- 11482130 TI - Apolipoprotein E polymorphism and Alzheimer's disease. AB - We aimed to determine the association and related factors of the apolipoprotein E (ApoE) genotype and Alzheimer's disease (AD) in Taiwan. We examined ApoE genotypes in 50 Chinese patients with AD and 50 age- and sex-matched controls. The patients met the criteria of probable AD of the National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA) and AD of the Diagnostic and Statistical Manual of Mental Disorders 4th edition (DSM-IV). There were 28 females and 22 males in the case and control groups. The mean age of onset of AD was 72. 62 years. The average interval between onset and research was 3.85 years. The frequency of ApoE epsilon 4 in the AD group was significantly higher than that in the controls (0.13 versus 0.02, p < 0.05). The odds ratio for AD in individuals with at least one ApoE epsilon 4 allele was 6.0 (95% CI 1.34 to 55.3, p < 0.001). The linear trend for AD in proportion to alleles of ApoE epsilon 4 was significant (chi 2 = 8.3, p = 0.004). The risk of ApoE epsilon 4 allele for the late-onset AD patients, males, or those who received less education was higher than that for the early-onset AD patients, females, or those who had received more education. The sensitivity of the epsilon 4 allele was 24%, the specificity 96%, the positive predictive value 86%, and the negative predictive value 56%. Our results supported that the ApoE epsilon 4 allele is related to AD in Taiwan. In addition, sex and education may play important roles in the presence of ApoE epsilon 4 allele. The epsilon 4 allele seemed helpful as an adjunct for diagnostic testing of AD. PMID- 11482131 TI - A missing data treatment for data mining applications in medical information systems. AB - To apply user-friendly, easily operated and accessible tools to handle missing data resulting from an auto-stored medical information system, these tools are applied to satisfy general users from different disciplines (i.e. statistics and machine-learning), followed by medical information system development. This study attempts to develop a new logic separation inference method applied to a database with a format like most real-world medical records containing many missing data and miscellaneous variables. It is expected that this method should have better performance than currently accessible methods. The newly developed logic separation inference method shows a classification power of 0.997 (elimination method is 1), which is better than the simple replacing method (replaced by mode shows 0.974). Both inference methods (mode and mean) have superior classification power to the simple replacing method. The missing data treatment processes introduced in this study can be completed on a MS Excel spreadsheet without any complicated calculation; therefore, they can satisfy general users. This new missing data treatment method is only applied up to 60% of the missing data (missing at random). However, when there is large amount of data, it is expected that this method also can be applied to a database missing more than 60%. PMID- 11482132 TI - Surgical treatment for concomitant fractures of the femoral neck and diaphysis: a treatment protocol. AB - Concomitant ipsilateral femoral shaft and neck fractures are difficult to treat. The standard protocol in our hospital is plates for diaphyseal fractures and lag screws or dynamic hip screws (DHS) fixations for the femoral neck fractures. We treated 21 patients with these complex fractures between 1988 and 1998 with this protocol. All patients were injured from high-energy trauma. There were 15 males and 6 females with an average age of 44 years. The average follow-up period is 50 months. All except one patient united well. During the follow-up period, neither non-union of the femoral neck fracture nor osteonecrosis of the femoral head was noted. There was also no malunion. There were twenty good results and one poor functional result. We conclude that plate on the shaft and sliding hip screws or separate screws in the hip are a reliable method for concomitant ipsilateral femoral neck and shaft fractures. PMID- 11482133 TI - Familial Mediterranean fever in a Taiwanese patient. AB - Familial Mediterranean fever is a rare disease characterized by cyclic attacks of fever, serositis and strong family background. Here we report a 22-year-old man who suffered from recurrent fever accompanied by chest and abdominal pain for more than 10 years. The attack frequency was about once per 2-3 weeks. Although he consulted many clinics and even received appendectomy at the age of 15, no definite diagnosis was given. During the admission, many laboratory examinations failed to show any abnormality except mild leukocytosis and elevated C-reaction protein. Image studies including chest X ray and abdominal CT scan showed negative result but, interestingly, Gallium-67 scan showed a hot spot in right lower chest and right lower abdomen. After prophylaxis with colchicine 1.0 mg per day, he has enjoyed more than 2 years without the above symptoms. PMID- 11482134 TI - Tarsal tunnel syndrome secondary to neurilemoma--a case report. AB - Peripheral nerve entrapment syndromes in the foot include those symptom complexes that are primarily neurologic in origin and result from embarrassment to any of the peripheral nerve trunks or branches of the foot. Tarsal tunnel syndrome usually is precipitated by compression of the tibial nerve posterior and distal to the medial malleolus. A neurilemoma is relatively uncommon in the foot. It is usually a solitary tumor that is almost exclusively benign and can be removed without jeopardizing the integrity of the nerve. Diagnosis is based on a thorough history and clinical pictures. Certain diagnostic modalities, ultrasound and MRI, have been employed to aid in diagnosis. Surgical excision of the tumor remains the treatment of choice. PMID- 11482135 TI - High resolution CT of Wegener's granulomatosis--case reports. AB - Computed Tomography (CT) of the chest was performed in three patients with Wegener's granulomatosis to define the anatomical basis of pulmonary involvement. Nodules, masses with cavitation, and areas of parenchymal opacities were typical manifestations on CT images. An extremely wide variety of radiologic findings were demonstrated in addition to the commonly described cavitary nodules. PMID- 11482136 TI - Root canal systems of the mandibular and maxillary first permanent molar teeth of south Asian Pakistanis. AB - AIM: The purpose of the present ex vivo study was to investigate variations in the root canal systems of mandibular and maxillary first permanent molar teeth of South Asian Pakistanis. METHODOLOGY: The root canal systems of a sample of 30 mandibular and 30 maxillary first permanent molar teeth extracted from South Asian Pakistanis were studied using a clearing technique. RESULTS: The mesial roots of the mandibular molar teeth typically presented with two canals (97%) of type II, IV or VI configuration. The distal roots of these teeth presented with a single canal (50%) of type I or V configuration or with two canals (50%) of type II, IV or VI. The prevalence of four root canals in two-rooted mandibular first permanent molar teeth was 47%. In maxillary molar teeth the mesial roots with a single canal (47%) were type I or type V; those with two canals (53%) were type II, IV or type VI. The distal and palatal roots that presented as a single canal (100%) were type I or type V configuration. The prevalence of four root canals in three-rooted maxillary first permanent molar teeth was 53%. CONCLUSION: It is concluded that four root canals in mandibular and maxillary first permanent molar teeth of South Asian Pakistanis is a common occurrence. The distribution of the different configurations of root canal systems in this population differed from that in Caucasian groups, suggesting that variations in root canal systems may be attributed to racial divergence. PMID- 11482137 TI - The efficacy of gutta-percha removal using ProFiles. AB - AIM: The purpose of this study was to compare the efficacy in vitro of gutta percha removal from obturated root canals using ProFiles. METHODOLOGY: Forty eight human root canals with curvatures ranging between 25 and 45 degrees were instrumented by a standardized method to an apical ISO size 30 and 0.04 taper. They were obturated with vertically condensed gutta-percha. Retreatment was performed with the following techniques: K-Flexofiles with chloroform; Hedstrom files with chloroform; ProFiles 0.04 taper with chloroform; ProFiles 0.04 taper alone. The time for each method was measured. A microfocal macroradiographic technique was used to evaluate the amount of debris remaining within the root canals after the retreatment procedure. Roots were divided into apical, middle and coronal parts and scored on a scale of 0 (no debris) to 3 (> 50% of walls covered with debris) by trained observers on two separate occasions. RESULTS: The scores for debris remaining within root canals for K-Flexofiles with chloroform and ProFiles with chloroform were the lowest and not significantly different at all three levels of the roots examined (P > 0.05), and Hedstrom files with chloroform and ProFiles with chloroform were not significantly different in the apical part. In general, coronal parts were cleaner than apical parts. The difference in scores at the three levels between ProFiles with chloroform and ProFiles alone were each significant (P < 0.01). Instrumentation using ProFiles with chloroform (mean 6.42 min) was significantly faster than using hand files (mean 11.67 min) (P < 0.01). CONCLUSION: The results indicated that ProFiles or hand files with chloroform produced similarly clean canals, but that ProFiles were faster. PMID- 11482138 TI - Three phosphor plate systems and film compared for imaging root canals. AB - AIM: The aim of this study was to compare three phosphor plate intraoral imaging systems (Digora (DA), Digident (DT), Denoptix (DX)) and E-speed conventional film (CF) for the imaging of root canals. METHODOLOGY: Sixty extracted permanent teeth were exposed using CF, DA, DT and DX. The length of root canal visible from its most apical extent to a line drawn at the level of the cementoenamel junction was measured and this was expressed as a percentage of the distance between the radiographic apex and the cementoenamel line. All images were examined concurrently by two examiners. Films were viewed under standardized conditions and DA, DT and DX images were viewed directly from the monitor screen. The images on the monitor were enhanced to give the subjectively clearest image. RESULTS: The mean percentage of canal visible on CF was 90%, DA 78%, DT 81% and DX 83%. The difference was significant for CF-DA (P < 0.001) and CF-DT (P = 0.001). There was no statistically significant difference for CF-DX or between any of the three phosphor plate imaging systems. CONCLUSIONS: It is concluded that a greater length of root canal was visible on conventional film than on three phosphor plate imaging systems and that this may be of clinical significance. PMID- 11482139 TI - Polymerase chain reaction detection of Treponema denticola in endodontic infections within root canals. AB - AIM: A 16S rDNA-based polymerase chain reaction (PCR) method was used to detect the occurrence of Treponema denticola in root canals. METHODOLOGY: Samples were collected from 54 single-rooted teeth having carious lesions and necrotic pulps. DNA extracted from the samples was amplified using the PCR assay, which yielded a specific fragment of T. denticola 16S rDNA. RESULTS: T. denticola was detected in 10 of 29 asymptomatic cases (34.5%), eight of 15 root canals that were tender to percussion (53.3%), and five of 10 teeth with acute periradicular abscess (50%). In general, T. denticola was found in 23 of 54 cases (42.6%). CONCLUSIONS: Findings suggest that T. denticola can be involved in the pathogenesis of periradicular lesions of endodontic origin. PMID- 11482140 TI - Periapical healing of mandibular molars after root-end sealing with dentine bonded composite. AB - AIM: The purpose was to present the frequency of periapical healing in first and second/third mandibular molars, after root-end resections sealed with a dentine bonded resin composite. METHODOLOGY: Root-end sealing of resected mandibular molar roots was made with dentine-bonded composite (Gluma-Retroplast) as a cover on the entire slightly hollowed root-end, in an attempt to prevent leakage. RESULTS: Out of the 834 roots recalled between 6 months and 12.5 years postoperatively, 92% showed complete apical healing, 1% uncertain healing and 7% exhibited failure. The healing result of 681 first molar roots was not significantly different (P = 0.21) from that of 153 second/third molar roots, and there was no significant difference in healing between mesial and distal roots (P = 0.32 for first molars, P = 0.86 for second/third molars) or amongst six age groups (P = 0.94). In the patient group: 71-89 years, 36 roots showed an average of 97% with complete healing. Out of 25 failures who were retreated surgically, 80% showed complete healing when examined subsequently. CONCLUSION: Root-end sealing of mandibular molars with dentine-bonded resin composite is a promising technique giving 92% complete healing in cases examined between 6 months and 12 years postoperatively. PMID- 11482141 TI - Coronal leakage following three obturation techniques. AB - AIM: To compare coronal bacterial and India ink leakage in three different obturation techniques with the smear layer having been removed. METHODOLOGY: Seventy extracted single-rooted teeth were instrumented to an apical preparation size 7 Profile Series 29 (Tulsa Dental Products, Tulsa, OK, USA). The smear layer was removed and 20 teeth were randomly obturated with lateral compaction, 20 teeth with vertical compaction, and 20 teeth with Thermafil (Tulsa Dental Products, Tulsa OK, USA). Ten teeth were used for positive (five teeth) and negative (five teeth) controls. Teeth were stored for 90 days in 100% humidity, then subjected coronally to Proteus vulgaris for 21 days to assess bacterial leakage. Following bacterial challenge, India ink was placed coronally for a further 21 days, then scored according to depth of dye leakage. RESULTS: Vertical compaction leaked significantly less than lateral compaction during bacterial challenge. However, when dye was used there were no significant differences. CONCLUSIONS: Bacterial leakage and dye leakage demonstrated considerable variability. The use of a dye following bacterial testing may highlight the failure of experimental devices and vertical root fractures, thus avoiding false positive results found with bacterial testing alone. PMID- 11482142 TI - An in vitro evaluation of the antimicrobial efficacy of irrigants on biofilms of root canal isolates. AB - AIM: The bactericidal effect of four antimicrobial agents was investigated against single-species biofilms derived from a range of root canal isolates. METHODOLOGY: Single-species biofilms of Prevotella intermedia, Peptostreptococcus micros, Streptococcus intermedius, Fusobacterium nucleatum and Enterococcus faecalis were generated on membrane filter discs and subjected to 15 min or 1 h incubation with 5 p.p.m. colloidal silver, 2.25% sodium hypochlorite (NaOCl), 0.2% chlorhexidine, 10% iodine or phosphate buffered saline (PBS) as a control. The antimicrobial activity of the agents was neutralized and the bacterial cells were harvested from the discs by vortexing, serially diluted in reduced transport fluid, plated on fastidious anaerobe agar containing 5% horse blood, incubated anaerobically and colony-forming units calculated. RESULTS: Iodine and NaOCl were more effective than chlorhexidine except against P. micros and P. intermedia where they were all 100% effective. Iodine and NaOCl elicited a 100% kill after 1 h incubation for all strains used. However, after 15 min, they showed differing bactericidal effects depending on the strain. None of the agents were effective against F. nucleatum after 15 min but NaOCl, iodine and chlorhexidine were all effective after 1 h. Colloidal silver was generally ineffective. CONCLUSIONS: The effectiveness of a particular agent was dependent on the nature of the organism in the biofilm and on the contact time. NaOCl was generally the most effective agent tested, followed by iodine. However the clinical efficacy of these agents must be considered in light of the complex root canal anatomy and polymicrobial nature of root canal infections. PMID- 11482143 TI - Effectiveness of oxidative potential water as a root canal irrigant. AB - AIM: The aim of this study was to evaluate the effectiveness of oxidative potential water (OPW) as an irrigant, based on its ability to remove the smear layer and/or debris from instrumented root canals. METHODOLOGY: One hundred and twenty root canals from extracted human maxillary incisors were instrumented using a conventional step-back technique with irrigation from sodium hypochlorite (NaOCl) or oxidative potential water (OPW). After instrumentation, the canals were irrigated by syringe or ultrasound using 15% EDTA or OPW as an irrigant. The volume of each irrigant used for syringe irrigation was 10, 20, and 30 mL, respectively, whilst the duration for ultrasonic irrigation was 1, 3, and 5 min, respectively. After irrigation, each root was split longitudinally in two with cutting pliers, and the specimens were prepared for SEM observation. The presence of debris and smear layer on each canal wall was assessed using a three-point scale for each parameter. RESULTS: Smear layer was effectively removed with EDTA both introduced via syringe and via ultrasonic irrigation. A similar effect was observed with OPW via syringe irrigation following instrumentation with 5% NaOCl. The canal walls in any of these cases showed open and patent dentinal tubules following smear layer removal. Some specimens irrigated with EDTA exhibited the effect of demineralization on the dentine resulting in funnelling of tubule orifices. Syringe irrigation was more effective in smear layer removal, except for ultrasonic irrigation with 15% EDTA, whilst ultrasonic irrigation was more effective in debris removal including the use of OPW as irrigant following instrumentation with 5% NaOCl. Neither syringe nor ultrasonic irrigation with OPW following instrumentation with OPW removed smear layer or debris effectively. CONCLUSIONS: The most effective irrigation technique for smear removal was 15% EDTA irrigation by means of syringe following instrumentation with 5% NaOCl solution. However, the most effective irrigation technique for debris removal was ultrasonic irrigation regardless of irrigant used. OPW irrigation by means of syringe following instrumentation with 5% NaOCl showed a similar effect to that of 15% EDTA irrigation for removal of smear layer and debris. PMID- 11482144 TI - Effect of Nd:YAG laser irradiation on the apical leakage of obturated root canals: an electrochemical study. AB - AIM: The purpose of this laboratory study was to evaluate the effect of Nd:YAG laser irradiation on the apical leakage of obturated root canals using an electrochemical method. METHODOLOGY: Forty extracted single-rooted teeth were selected and the anatomic crown of each tooth was removed. The specimens were randomly divided into four groups. In group 1, the root canals were prepared with K-files and irradiated with Nd:YAG laser (5 W, 20 Hz) via a 300 microns optical fibre. Then the root canals were obturated with laterally condensed gutta-percha and Pulp Canal Sealer EWT. In group 2, the root canals were treated with the same method as those of group 1 but without laser irradiation. In group 3, the root canals were prepared with ProFiles, laser irradiated and then obturated with vertically condensed gutta-percha and Pulp Canal Sealer EWT. In group 4, the root canals were treated with the same method as those of group 3 but without laser irradiation. The electric resistance between standard and experimental electrodes in the canals was measured over a period of 10 days. RESULTS: At 2 h, groups 1 and 3 irradiated with laser had significantly less apical leakage than group 2 (P < 0.05). After 10 days, group 4 had the highest leakage, followed by groups 2, 1 and 3; the differences between the groups was statistically significant (P < 0.05). CONCLUSIONS: Laser irradiation following root canal preparation reduced apical leakage following root canal obturation. PMID- 11482145 TI - The influence of preparation technique and sodium hypochlorite on removal of pulp and predentine from root canals of posterior teeth. AB - AIM: To test the null hypothesis that removal of pulp tissue and predentine from root canals of multirooted teeth is not significantly different when performed by one of two mechanical preparation techniques in conjunction with either water or 3% sodium hypochlorite (NaOCl) irrigation. METHODOLOGY: Forty-six freshly extracted premolar or molar teeth, with pulps vital at the time of extraction, were assigned to four groups balanced by anatomy (group 1, step-back filing/NaOCl; group 2, step-back filing/water; group 3, automated rotary/NaOCl; group 4, automated rotary/water). Preparation was undertaken by a single operator and the volumes of irrigant recorded for each tooth. One root from each tooth was prepared for transverse histological assessment at eight levels, including one at and two apical to the working length. A validated index was used to measure the presence and location of residual pulp tissue and predentine. RESULTS: No statistically significant differences were found in the frequency with which residual pulp tissue or predentine were observed amongst the groups. Coronal parts were more frequently devoid of pulp tissue and predentine than the apical. Canals were frequently occluded with residual pulp tissue apical to the working length. There was a trend for better pulp and predentine removal in accessible parts of the canal as a function of rotary preparation and NaOCl irrigation. The isthmus had residual pulp tissue less frequently in the NaOCl groups. Accessory anatomy was unaffected regardless of technique or irrigant. CONCLUSIONS: Pulp tissue and predentine removal were not significantly different between a step back filing and an automated rotary preparation technique in conjunction with water or NaOCl. PMID- 11482146 TI - Root canal treatment in mandibular canines with two roots: a report of two cases. AB - AIM: To highlight anatomical variation in the mandibular canine. SUMMARY: Two clinical case reports are presented to illustrate anatomical variation in the human mandibular canine. Endodontic treatment may sometimes fail because morphological features of the tooth adversely affect the treatment procedures. Many investigators have reported the anatomical variations associated with mandibular canines. Mandibular canines are recognized as usually having one root and one root canal in most cases, although approximately 15% may have two canals or sometimes two roots. This paper describes two clinical cases of mandibular canines with two roots and two canals. KEY LEARNING POINTS: Failure to control infection in the root canal system has an adverse impact on outcome. Clinicians should be aware of anatomical variations in the teeth they are managing, and should never assume that canal systems are simple. The majority of mandibular canines have one root and root canal, but 15% may have two canals, and a smaller number may have two distinct roots. PMID- 11482147 TI - [Surgery of esophageal achalasia--possible complications]. AB - The authors demonstrate on a group of patients operated by a miniinvasive method possible complications of treatment of achalasia of the oesophagus. On a group of 61 patients they analyze all complications and try to detect their causes. They divide complications into peroperative, early and late postoperative ones. The most frequent peroperative complications are perforation of the oesophagus which are as a rule treated by the laparoscopic route. After surgery the most frequent problem is a relapse of achalasia. The latter is sometimes due to inadequate myotomy. This can be prevented by careful surgical technique along with peroperative endoscopic control. PMID- 11482148 TI - [Carcinoid of the ileocecal junction with symptoms of acute abdomen]. AB - The authors describe a group of six patients (two men and four women) operated at the Surgical Clinic in Plzen during 1995-2000 on account of a carcinoid of the GIT. The mean age of the patients was 48.6 years, the range 14-77 years. In five instances a carcinoid of the appendix was detected. In a separate case-record they evaluate a carcinoid of the ileocaecal transition with symptoms of acute abdomen, early surgery, its extent incl. the postoperative procedure. PMID- 11482149 TI - [Serious surgical complications associated with chronic anticoagulant therapy]. AB - Chronic anticoagulant treatment is administered mostly for cardiological reasons. Cumarin derivatires are used in the majority of cases (Warfarin, Pelentan). It is necessary to monitor this treatment regularly and to control the dose according to the INR value. Different complications can occur; the haemorrhage represents a serious one. The authors discuss several aspects of anticoagulant therapy and possible prevention of the complications. The importance of the problems is demonstrated on the authors' clinical experience--two cases of haemorrhage after Warfarin administration simulating an acute surgical event. PMID- 11482150 TI - [An unusual extensive thrombus in the abdominal aorta of a young female patient after recurrent embolisms in the pelvis and lower extremity]. AB - The authors present a case of an uncommon based thrombus of the abdominal aorta, which partly obturated its lumen and caused repeated embolization of the left pelvic circulation and left lower extremity. Even detailed modern diagnostic methods (angiography, CT, NMR) were unable to rule out before surgery anotheretiology of the obturating process in the aorta. Only removal of the thrombus from the abdominal aorta and subsequent histological examination elucidated the diagnosis and led to final cure. PMID- 11482151 TI - [Excision of extrahepatic bile ducts in laparoscopic cholecystectomy]. AB - The authors describe the case of an excision of the extrahepatic portion of the right, left and common hepatic duct. Concurrently the authors discuss indications for peroperative drainage of biliodigestive anastomoses. At the Third Surgical Clinic the authors operated during the last three years nine patients with injuries of the biliary pathways during laparoscopic cholecystectomy. In three patients it was possible to suture partial injuries to a T-drain. In six patients with complete severing of the pathways a hepatoenteral anastomosis was established. PMID- 11482152 TI - [Dynamics of leptin plasma levels after abdominal surgical procedures]. AB - In addition to its basic role as regulator of the amount of body fat and food intake leptin plays a part also as an acute phase reactant. A rise of the leptin level during this period may be associated with anorexia and cachexia in relation to general inflammatory manifestations. The objective of the present study was to characterize the dynamics of the leptin concentration in patients after intraabdominal surgery. PATIENTS AND METHODS: The prospective study was implemented in a group of 16 men subjected to planned surgery--resection of colorectal carcinoma. The plasma leptin concentration (RIA), TNF alpha, IL-beta. IL-6, sIL-2R (ELISA), CRP, alpha-1-antitrypsin, alpha-1-acid glycoprotein and ceruloplasmin (nephelometry) were assessed before surgery and then +24, +48 and +72 hours after the beginning of the operation and compared with concentrations in healthy subjects. RESULTS: Plasma leptin concentrations culminate +24 hours after surgery and in this stage they do not correlate with the BMI. Within 48 hours the concentration reaches rapidly normal levels. TNF alpha, IL-6 and sIL-2R reach maximal levels 24-48 hours after the onset of surgery with a subsequent slow decline. All acute phase proteins (APP) have prolonged dynamics with an elevation within +72 hours. A significant correlation was proved between the leptin concentration and TNF alpha 24 hours after surgery (r = 0.43, p < 0.05) and between leptin and IL-6 24 hours after surgery (r = 0.32, p < 0.05). CONCLUSION: The dynamics of the plasma leptin concentration during the postoperative period differ from the typical development of early indicators of the systemic inflammatory response--cytokines as well as from the dynamics of APP. Leptin elevation during this period may contribute to anorexia of patients after major surgery. However, rapid normalization of the level within 48 hours after surgery indicates however that leptin is involved also in other, so far unspecified factors with an anorectic action. PMID- 11482153 TI - [Videothoracoscopy and video-assisted surgical procedures in penetrating injuries of the thorax]. AB - Diagnosis and treatment of the penetrating injury of the chest is quite difficult. In all types/sizes of hospitals/these injuries has to be immediately treated in surgery departments. Often decision about the optimal treatment of these injuries is quite difficult. In this paper experience with 37 cases of penetrated trauma is presented. Authors defined adequate criteria for selection of the chest tube, videothoracoscopy, video-assisted thoracic surgery (VATS), emergency thoracotomy and thoracophrenolaparotomy. PMID- 11482154 TI - [A new method of therapy of hemorrhoids using the PPH stapler and the Longo technique]. AB - The author wish to submit information on the use of a new method of surgical therapy of haemorrhoids about which they acquired information at a workshop of Ethicon Co. in Ljubljana (Slovenia). At the same time they want to demonstrate their first experience with introduction of the new method into practice. PMID- 11482155 TI - [Laparoscopic fenestration of lymphoceles after kidney transplantation]. AB - Lymphocele is a relatively frequent complication after kidney transplantation, which impair the passage of urine from the kidney, cause emptying of bladder lymphoedemas of the lower limbs, etc. Percutaneous drainage of lymphocele is associated with a risk of infection and a high percentage of recurrence. Until recently, standard surgical treatment of this complication was open fenestration of the lymphocele into the peritoneal cavity. In their paper, the authors describe their first experience with laparoscopic fenestration of the lymphocele. From May 1998 till April 1999 the authors performed laparoscopic fenestration in 5 patients. In four patients the intervention was successful, in one female patient recurrence of lymphocele was observed, which was later resolved by an open operation. In none of the patients early or late surgical complications occurred, the use of analgetics during the postoperative period was minimal, the length of hospitalisation ranged from 2 to 4 days. The authors consider the discussed method a suitable alternative for treating lymphocele after renal transplantation thanks to its minimal invasivity. It involves practically no load of the sick patients after kidney transplantation. PMID- 11482156 TI - [Surgical treatment of thyroid diseases at the Surgical Clinic of the University Hospital of Plzen 1994-2000]. AB - The authors operated in 1994-2000 at the Surgical Clinic, Faculty Hospital Plzen a total of 793 patients on account of thyroid disease. This group comprised 57 patients with malignant thyroid disease. The mean age of the operated patients was 48.8 years (range 12-89 years). Three patients (0.38%) died within 30 days after operation. The postoperative morbidity was 6.1%. A lesion of the recurrent nerve was recorded during the postoperative period in 24 patients (3.03%) and had a declining trend. Postoperative hypoparathyroidism was recorded in 5.8% (n = 46) injuries of the neighbouring organs in 1.02% (n = 8), oedema of the larynx in 18 patients, i.e. 2.3%. On the whole the authors observed a decline of postoperative complications during the investigation period, the number being lowest in 2000 associated with the number of performed thyroid operations and a change of the surgical strategy. PMID- 11482157 TI - [Occurrence of yeast infections at a surgical intensive care unit]. AB - The authors evaluate the occurrence of candida infections in the surgical intensive care unit (ICU) in a one-year period. From the group of 1798 patients hospitalized in a surgical ICU there are 50 patients (2.8%) with candida colonisation and even 56 patients (3.1%) with candida infection. The authors evaluate risk factors for development of candida infection, its etiologic agents and the most frequent site of infection. In addition to usual treatment the authors suggest pre-emptive therapy in patients with risk factors. In the one year period, two patients died of multiorgan failure (MOF) in close connection with candida infection (sepsis). The mortality of the group with proved candida infection was 3.57%. PMID- 11482158 TI - [Surgical treatment of upper gastrointestinal hemorrhage of peptic ulcer origin not treatable by endoscopy]. AB - The authors present possibilities of treatment of an acute bleeding peptic ulcer in the upper part of the gastrointestinal tract. They give options of surgical treatment in case of failure of an endoscopic approach. They evaluate results of endoscopic and surgical treatment in the retrospective of a five-year study and compare the benefit of different kinds of operative interventions. PMID- 11482159 TI - HIV/AIDS awareness among hospital employees. AB - The present cross sectional study was carried out to assess the basic knowledge and attitude of class III and Class IV hospital employees (n = 250) regarding HIV/AIDS. Of the 250 study subjects 232(92.8) had heard or read about AIDS. Chief sources of information were television, newspapers and doctors. Though the study subjects had some factual knowledge about HIV/AIDS, certain misconceptions and myths persisted. They were also found to have a biased and negative attitude towards people with AIDS. 223(96.1%) study subjects said that they would like to learn more about HIV/AIDS. The results of this study indicate that action is needed to increase the level of knowledge of this group so that they reject biased information and myths relating to AIDS and thus carry out their duties effectively and help in forming an enlightened opinion against AIDS. PMID- 11482160 TI - Seroprevalence of cytomegalovirus infection in Mauritian volunteer blood donors. AB - OBJECTIVE: To determine the seroprevalence of CMV antibodies in the Mauritian volunteer blood donor population and to establish a panel of CMV-seronegative blood donors. STUDY SUBJECTS AND METHODS: Five hundred and eighty four apparently healthy blood donors were screened for evidence of CMV infection by the complement fixation test. There were 551 males and 33 females with age ranging from 18 to 60 years. RESULTS: Complement-fixing antibodies were found in 93.5% of the blood donors. The prevalence was 93.1% in males and 100% in females. CONCLUSION: Our findings demonstrate that seroprevalence of CMV in the local blood donors is very high making CMV-seronegative blood very scarce. Therefore leucocyte-depleted blood should be used as an alternative to CMV-seronegative blood during transfusions. PMID- 11482161 TI - Lipid peroxidation and antioxidant enzymes in coronary and peripheral artery disease. AB - There is a strong co-relation in between the levels of blood lipids in patients of different subgroups of CAD, however this is not present in the case of subgroups of patients in PAD. Moreover levels of MDA and antioxidant enzymes are also significantly altered in the subgroups of CAD but the correlation is weak in that of PAD. Thus these values may serve as a clinical support for experimental data and supplementary information regarding atheromatous disease. PMID- 11482162 TI - Ginger, fat and fibrinolysis. AB - Administration of 50 gm of fat to 30 healthy adult volunteers decreased fibrinolytic activity from a mean of 64.20 +/- 5.31 to 52.10 +/- 3.20 units (P < 0.001). Supplementation of 5 gm of ginger powder with fatty meal not only prevented the fall in fibrinolytic activity but actually increased it significantly (P < 0.001). This fibrinolytic enhancing property is a further addition to the therapeutic potential of ginger. PMID- 11482163 TI - Management of toxoplasmosis in AIDS. PMID- 11482164 TI - Health problems of tribal population groups from the state of Maharashtra. PMID- 11482165 TI - Acrosome intactness and seminal hyaluronidase activity: relationship with conventional seminal parameters. AB - Seminal hyaluronidase activity was estimated after liquefaction in semen samples of 100 male partners of infertile couples including 16 azoospermic (no spermatozoon) men and 48 fertility proven men by a method based on measurement of the area of digestion of substrate (hyaluronic acid) in agar plate. Semen samples were also evaluated for Acrosomal Intactness (AI) test except the azoospermics of the studied samples. Seminal hyaluronidase activity was completely absent in azoospermic specimens confirming its cellular origin. Seminal hyaluronidase activity was found to be significantly correlated, statistically, with sperm density (r = 0.708, p < 0.001), % motility (r = 0.6478, p < 0.001) and % normal sperm morphology (r = 0.5724, p < 0.001). Acrosomal Intactness (AI) test scores were also well correlated with sperm density (r = 0.6477, p < 0.001), % motility (r = 0.5965, p < 0.001) and % normal morphology (r = 0.6237, p < 0.001). Both values were higher in semen samples with normal routine parameters (proven fertility and normozoospermic infertile groups) than those compared with abnormal routine parameters (oligozoospermic). We also found very highly significant correlation (r = 0.8442) between seminal hyaluronidase activity and Acrosomal Intactness scores, statistically (p < 0.001). This could be because; normal germinal semineferous epithelium generates abundant number of sperms with normal motility and morphology that are also having intact acrosome. Intact acrosome prevents loss of acrosomal enzymatic activity (e.g. hyaluronidase) until released after liquefaction during seminal analysis and during acrosomal reaction in female genital tract prior to fertilization. Seminal hyaluronidase activity, thus determined, is primarily dependent upon the intact status of acrosome. As each sperm contributes to the seminal hyaluronidase activity, it is directly correlated with sperm density; but at the same time it exhibits goods correlation with % motility and % normal morphology. Therefore AI score and seminal hyaluronidase activity can be considered as good indicators of sperm function. PMID- 11482166 TI - Lipid peroxidation and diabetic retinopathy. AB - The high level of glucose in blood for a long duration is the main cause of the development of retinopathy. So yearly screening of patients newly diagnosed with NIDDM diabetes is recommended because rare cases of treatable diabetic retinopathy have occurred early in one course of NIDDM diabetes. Hyperglycaemia leads to non-enzymatic glycosylation of proteins and HbA1C was found increased. Antioxidants such as GSH and SOD level is found decreased in retinopathy conditions due to the higher lipid peroxidation, which is evident from high MDA and DC values. So it can be clearly stated that increase in the free radical by hyperglycaemia, lipid peroxidation and advanced glycosylation endproducts along with decreased antioxidants are the causative agents for the development of retinopathy. PMID- 11482167 TI - Antidepressants induced sexual dysfunctions. PMID- 11482168 TI - Seroepidemiology and active surveillance of dengue fever/dengue haemorrhagic fever in Delhi. AB - The aims of the present study were to carry out surveillance for dengue virus infection in adults with short-duration fever, and serological study of dengue virus infection in persons without fever. Patients were divided into two groups. Group 1 included patients above 12 years of age with fever of 2-12 days duration without any apparent cause. Of these, patients who presented with fever for 2-5 days were included for virus isolation (group 1a) while those who presented within 6-12 days of the onset of fever were included for the dengue specific IgM serology (group 1b). Group 2 included a sample of population belonging to all age groups but without pyrexia and blood was collected for dengue-specific IgG serology. Twenty-six patients were enrolled in group 1a over a period of 4 months (September to December, 1997). Of these, DEN1 was isolated in 5 cases. Group 1b included 182 patients, out of which 34 (18.68%) were positive for dengue-specific IgM antibodies. Significantly, all the positive cases were detected during the months of September to November. Retro-orbital pain was present in a significantly more number of IgM-positive cases as compared to IgM-negative cases. Group 2 included 125 cases without fever. The overall positivity for dengue-specific IgG antibodies was 77.6%, with the highest positivity of 100% in the age group of 31-40 years. It was concluded that dengue virus infection is endemic in and around Delhi with peak incidence between September and Novemver. The prevalent serotype during September and December 1997 was DEN1. Since previous epidemic of DHF was due to DEN2 type, isolation of DEN1 serotype indicates changes of another epidemic of DHF due to DEN1 serotype. The stresses the urgent need for implementation of measures to control the transmission of dengue infection. PMID- 11482169 TI - Failure of oral iron therapy in treatment of iron deficiency anemia: pharmaceutical iatrogenic cause. PMID- 11482170 TI - Malignancy in AIDS: institutional management or home care? AB - In India, the first sero-positive person was reported from Chennai in 1986 and the first case of AIDS from Mumbai 1987. Since then the epidemic is growing steadily and WHO/UNAIDS estimate that by the year 2000 AD India has around 3-5 million people suffering from HIV/AIDS and this is the largest burden of HIV in single country. Over 40% of all patients with HIV infection will develop malignant disease at sometime during their course of illness. AIDS and malignancy together constitute a deadly combination. The authors recommend that the care of patients having malignancy with AIDS should be mainly at home rather than in an oncology center. This will ensure the patient's and family's comfort as most of the specialized cancer centers in India are at a considerable distance away from patient's homes. Frequent visit to a cancer center will exhaust the patient and attendants emotionally, physically and financially. This is especially true because irrespective of the pains taken by the family and the specialists, a cure is an unlikely possibility. Only one specialist from oncology and one primary health physician, who are completely aware of the patient's physical, psychological, financial and social background, should be responsible for the care of a patient having malignancy with AIDS in India. The main advantages of the home care of such patients are: patient's and family's comfort; lesser mental, physical, social, emotional and financial agony; and minimum need for institutional care, which may be instituted as and when necessary. PMID- 11482171 TI - Functions of endothelium. PMID- 11482172 TI - Anti-leukaemia cell. PMID- 11482173 TI - Capacity considerations and community benefit expenditures of nonprofit hospitals. AB - The United States Internal Revenue Service (IRS) and some states require nonprofit hospitals to demonstrate that they provide a substantial community benefit in order to get or maintain their tax-exempt status. This places every nonprofit hospital at risk. This article is about certain common factors that impact the management of all nonprofit hospitals and their ability to comply with such laws. PMID- 11482174 TI - Rural residence and migration for specialty physician care. AB - This article describes why rural residents migrate or travel outside their local market area for specialty physician care. Data were collected through a random mail survey of persons residing in Iowa's rural counties. The results imply that migration for specialty care is not simply a function of a low perceived availability of local specialty physicians. Managers of rural and urban health care systems may need to rethink the extent to which specialty physician services should be distributed across rural markets. PMID- 11482175 TI - Implications for health care management. PMID- 11482176 TI - External threats and nursing home administrator turnover. AB - External threats and volatility in the long-term-care sector in recent decades have posed serious challenges for nursing home administrators. Greater job complexity and administrative responsibilities resulting from public policies and more specialization and competitiveness in nursing home markets have made turnover a significant issue. This article examines administrator turnover from 1970 through 1997 in New York State and describes how turnover increased markedly in the late 1980s and early 1990s. PMID- 11482177 TI - A comparison of the positive and negative work attitudes of home health care and hospital nurses. AB - This article compares the positive and negative stress response work attitudes of 126 home health care registered nurses (RNs) and 145 hospital RNs. Home health care nurses reported significantly lower levels of the negative work attitudes, anger/hostility, and frustration with the system. There was no significant difference between the two groups of RNs among the positive work attitudes of job satisfaction, positive affect, hope, meaningfulness, manageability, and trust in the supervisor and the negative work attitudes frustration with the supervisor, job alienation, and negative affect. PMID- 11482178 TI - The impact of workplace empowerment, organizational trust on staff nurses' work satisfaction and organizational commitment. AB - A predictive, nonexperimental design was used to test Kanter's work empowerment theory in a random sample of 412 staff nurses selected from the professional registry list of a central Canadian province. Kanter argues that work environments that provide access to information, support, resources, and opportunity to learn and develop are empowering and influence employee work attitudes, productivity, and organizational effectiveness. Test results suggest that fostering environments that enhance perceptions of empowerment will have positive effects on organizational members and increase organizational effectiveness. PMID- 11482179 TI - A comparative study of resident, family, and administrator expectations for service quality in nursing homes. AB - This article compares the service quality expectations of residents, family, and administrators within a long-term-care context. Expectations for service quality are measured using the SERVQUAL scale. The results support the reliability of the SERVQUAL instrument for nursing home residents. Resident expectations were significantly different from administrators' expectations on all dimensions of quality. The findings are interpreted in terms of expectation formation and the nature of service encounters. PMID- 11482180 TI - Hospital diversification into long-term care. AB - In the 1990s, acute care hospitals in the United States encountered an unstable operating environment created by a series of transformations in the health care delivery system and long-term-care market. Confronted with an array of economic pressures and demographic changes, hospitals were motivated to engage in long term-care diversification, such as establishing a long-term-care unit or providing home health services, as a means of entering new markets and ensuring financial stability. This article examines the organizational, market, and community factors associated with this strategic activity among a national sample of urban and rural hospitals. PMID- 11482181 TI - [Angiographic characteristics of the vertebral arteries stenosis in the cerebral blood circulation insufficiency of vertebrobasilar pool]. AB - The angiography (AG) is the most important and obligatory part of diagnostic complex. The AG data characterizes mostly full the type and hemodynamical features of the vertebral arteries (VA), which are presented by typical inborn and acquired changes of V1 and V2 segments. Stenotic affection of VA constitutes the indication to surgical treatment. PMID- 11482182 TI - [Application of puncture chemical desympathization under the CT control in the treatment of chronic arterial insufficiency of extremities]. AB - For elimination of sympathetic influence on the extremities vessels in obliterating disease of their arteries in 126 patients the desympathization was done, including 66 causes of punctural paravertebral introduction of sympathicoleasing substances under the computer tomography (CT) control and in 60 -using sympathectomy according to Leriche or Texatsu. The lower traumaticity and efficiency of the chemical desympathization method under the CT control permits to recommend it as alternative to reconstructive intervention or surgical the sympathectomy, in patients with significant of current diseases in particular. Application of method had permitted to reduced the duration of stationary treatment of patients to 1.5 days. PMID- 11482183 TI - [Surgical treatment of occluding lesion of the subclavian artery]. AB - Results of cerebral hemodynamics investigation and operative treatment of 53 patients with the subclavial artery (SA) occlusion were presented. It was established that the course of the cerebral vascular disorders occurrence in patients with the SA proximal segment occlusion is the vertebro-subclavial robbing syndrome. The consider that the SA resection with her implantation to ipsilateral a. carotis communis (ACA) is operation of choice. The vessel prosthesis is indicated in presence of widespread affection. It is expedient to performed one-stage correction of blood flow in the vessels mentioned in patients with ipsilateral localization of the SA and ACA affection. PMID- 11482184 TI - [Minimally invasive method of treatment of hepatic abscesses]. PMID- 11482185 TI - [Purulent-necrotic complications of acute pancreatitis]. PMID- 11482186 TI - [Inflammation response syndrome, sepsis and multi-organ insufficiency in patients operated for destructive pancreatitis]. AB - In 128 patients, operated on for destructive pancreatitis (DP), the systemic inflammatory response syndrome (SIRS), sepsis and polyorganic insufficiency (POI) was investigated. SIRS was revealed in 56.3% of observations, the severe one--in 31%, sepsis--in 39.8%, including in 79.4% reoperated patients. According to bacteriological investigation data the diagnosis of sepsis was confirmed in 26.3% of patients only. POI was revealed in 25% of patients, including in 59.4% of those, who were reoperated. PMID- 11482187 TI - [Prognostic significance of immune reactions in multiple injuries involving thoracic and abdominal cavities]. AB - Prognostic informativity some indexes of immune response in severe shockogenic combined thoracoabdominal trauma was studied. In 92 patients aged from 17 to 85 years (68 men and 24 women) in acute period of traumatic disease (TD) quantity of T-, B-, 0-lymphocytes, immunoregulating subpopulations, the main immunoglobulins classes concentration, circulating immune complex were determined. Significant disorders of the organism immune system of injured persons were revealed, but prognostic significance of the data obtained in determination of the TD course severity was not large. PMID- 11482188 TI - [Surgical strategies in combined injury of pelvic bones and organs]. AB - Experience of surgical treatment of the damaged pelvic bones and its organs in 124 injured persons was summarized. The authors consider, that in combined trauma of pelvis and its organs it is necessary to undertake maximally possible reanimational-surgical measures during 12-72 hours after trauma occurrence, depending on its severity, character of injury of pelvic organs and bones. External fixation is the method of choice in unstable fracture of pelvic bones in early period of traumatic disease. Mortality was 10.5% (13 injured persons). Application of an active surgical tactics in early period of traumatic disease had promoted the shortening of the infectional complications frequency as well as of duration of stationary treatment in 2 times. PMID- 11482189 TI - [The influence of intra-abdominal electrophoresis and enteral sorption on the immune indices in children with peri-appendicular abscess]. AB - Comparative characteristic of application of different methods of intratissue electrophoresis and their influence on immune indexes in children with perioappendicular abscess was presented. Application of complex treatment with usage of intraabdominal electrophoresis and enterosorption had guaranteed an effective correction of systemic and local immune disorders, the peritoneal macrophages activation, improvement of cytokines cooperation of cells in the immune answer. PMID- 11482190 TI - [Experience of surgical treatment of thymoma]. AB - For period from 1992 to 2000 years 72 patients with thymoma were operated. In 29 (40.2%) patients lymphocytic thymoma was revealed, in 10 (13.8%)--"benign". Postoperative mortality was 4.1%. In term from 1 to 1.5 years 5 (6.9%) patients died. PMID- 11482191 TI - [Possibilities for surgical treatment of locally spread head and neck cancer]. AB - The results of treatment of 263 patients with locally spread cutaneous cancer of head and neck, lower lip, the oral cavity mucosa, larynx and thyroid gland, to whom 299 surgical interventions were done, including 121 radical excision of cervical fat with prophylactic and curative aim, were analyzed. There were admitted to hospital 187 (71.1%) patients with III stage of the disease 41 (15.6%)--in IVa stage, with tumor recurrency--35 (13.3%). Common postoperative complications occurred in 19 (7.2%) patients and the local--in 83 (31.6%). The best 5-year survival indexes were noted in cutaneous and thyroid gland cancer (69.8 and 70.5% accordingly); the poorest--in cancer of the oral cavity mucosa (39.7%); in cancer of lower lip and of larynx--52.9 and 61.3%. PMID- 11482192 TI - [Histological structure of a tumor as a criterion of primary multiplicity in bilateral cancer of mammary glands]. AB - Comparative estimation of histological structure of contralateral tumors in 104 patients with primary and in 54--with metastatic mammarial gland cancer (MGC) was performed. Revealing of various histological types of tumoral nodes--of a one component infiltrating ductal cancer (IDC) in one mammarial gland and a one component infiltrating lobular cancer (IPC)--in another one, combination of special form of the MGC with IDC and/or IPC, combination of cancer in situ with invasive form of the MGC constitutes the criterion of primarily-multiple affection. In differentiated diagnosis of primary and metastatic tumor this criterion is highly specific (92.9%), but has low prognostic value (37.5%). That's why in 73% of women-patients it is necessary to analyze the clinical examination data in addition to establish the diagnosis of primary bilateral MGC. PMID- 11482193 TI - [Fundamental s of anesthesiology-reanimation. Part II. Principles of training]. AB - The patient's reconvalescence, who fall ill suddenly or is injured or operated on, is impossible without the organism biological integrity (OBI) preservation or restoration and this constitutes the essence of the anesthesiology-reanimatology (AR) subject. The OBI is regarded as a process of necessary realization for the morphostructural balance preservation and of homeokinetic interaction of biological and energodynamical potential of organism. The OBI weak chains are determined, which are responsible for the course and the progression rate of bioenergetic insufficiency, acidotic alteration and the morphostructural balance disorder. The technology is adduced for the injuries elimination using the system of the local integrity restoration and the restoration OBI system. The AR application would raise the clinician's methodological armamentarium and promote to improve the result of treatment of patients in critical state. PMID- 11482194 TI - [Prospects for differentiated conservative treatment of patients with benign prostatic hyperplasia]. AB - The results of conservative therapy of 30 patients aged from 50 to 79 years with benign hyperplasia of prostate according to the IPSS scale from 7 to 30 points were analyzed. Under the influence of treatment in all the patients the clinical symptoms severity, according to the monitoring data using the IPSS tables, had reduced by 72% at average and quality of life improve no less than by 50%. Average efficacy of the volume and volumetric of urination had enhanced according to uroflowmetry data. PMID- 11482195 TI - [Reconstructive-restorative operations in rehabilitation of patients after colonic damage]. PMID- 11482196 TI - [Venous thromboembolic complications in abdominal and vascular surgery. The risk factors and measures of prophylaxis]. AB - There were analyzed 4675 histories of cases of patients, who were operated on for disease of the digestion organs (3349) and for obliterating atherosclerosis of abdominal aorta and peripheral arteries (1326). The risk factors of occurrence were established, the prophylaxis methods for venous thromboembolic complications (TEC) in postoperative period were elaborated. High efficacy of application of fraxiparine in the venous TEC prophylaxis in abdominal and vascular surgery was noted. PMID- 11482197 TI - [Application of own cells of an organism for the medicinal remedies transport]. PMID- 11482198 TI - [The method of prognosis of acute pancreatitis]. PMID- 11482199 TI - [Laparoscopic cholecystectomy in situ for inverted viscera]. PMID- 11482200 TI - [Combined palliative subtotal pancreatectomy with resection of two hollow organs in cancer surgery]. PMID- 11482201 TI - [Rare observation of nonspecific ulcerative colitis]. PMID- 11482202 TI - [External jugular vein: alternative vascularized access]. PMID- 11482204 TI - [On the 80th anniversary of the Odessa Surgical Society (a brief history)]. PMID- 11482203 TI - [The involvement of the entire genitourinary tract in neurofibromatosis in a child]. PMID- 11482205 TI - [Clinical value of determination of the blood elastase activity as a marker of postoperative pancreatitis]. AB - For diagnosis of postoperative pancreatitis 50 patients were examined after performance of organ-preserving operation for gastroduodenal ulcer disease. In 15 patients various postvagotomy symptoms occurred. In 12 of them the elastase activity in blood was increased already in 12 h after operation, lowering gradually under the influence of specific therapy. On 2-3 days in these patients the amylase activity increase in blood and urine was revealed, testifying the postoperative pancreatitis presence. Under the influence of the treatment conducted the postvagotomy syndromes severity had reduced in accordance with the elastase activity normalization in blood. PMID- 11482206 TI - [The influence of perftoran on the course of acute pulmonary injury syndrome in patients with destructive pancreatitis]. AB - The respiration disorders are revealed in patients with an acute destructive pancreatitis in early and late terms of the disease. Occurrence of an acute pulmonary injury syndrome (APIS) worsens the disease prognosis, enhances mortality. In complex of intensive therapy the perftoran (Russia) infusion in 5 ml/kg of body mass dose are applied in 10 patients with destructive pancreatitis. Main indexes of the blood gases composition and of hemodynamics were controlled. It was established that application of perftoran detains the APIS occurrence in the patients with severe pancreatitis, averts the polyorgan insufficiency occurrence, promotes the various kinds of hypoxia correction, stimulates cardiac activity, raises the blood oxygen capacity, improves microcirculation. Application of perftoran was substantiated in complex of treatment of patients with an acute destructive pancreatitis for prophylaxis of occurrence and for elimination of APIS. PMID- 11482207 TI - [The influence of clexane on the fibronectin content in the blood plasma in patients with destructive pancreatitis]. AB - The fibronectin content dynamics in the blood serum was estimated in patients with various clinical forms of an acute pancreatitis. The fibronectin level lowering may be a predictor of occurrence and diagnostical criterion for infective complications in patients with destructive pancreatitis. Application of clexane had promoted the fibronectin level raising in the blood serum in patients with sterile and infected pancreatic necrosis. PMID- 11482208 TI - [Variants of surgical treatment of cholecystitis]. AB - Basing on analysis of the treatment results of 375 patients with cholecystitis the indications for application of broad laparotomy, variable laparotomy, mini access, endotracheal narcosis and epidural anesthesia are substantiation. PMID- 11482209 TI - [Laparoscopic cholecystectomy late follow-up results and quality of life of patients]. AB - Quality of life of 198 patients in term for 1 to 3 years after performance of laparoscopic cholecystectomy (LCHE) was studied. The questionnaire data were estimated according to scale of Visick. Late follow-up result was excellent (Visick I) in 81 (40.9%), good (Visick II)--in 45 (22.7%), fair (Visick III)--in 53 (26.8%) and bad (Visick IV)--in 19 (9.6%) of patients. Patients with fair (Visick III) and bad (Visick IV) results were examined to reveal the cause of such outcome. The main causes of fair result were chronic disorders of gastroduodenal motor-evacuation function and of the bad result--noncorrected residual and iatrogenic traumatic pathological changes of extrahepatic biliary ducts. The quality of life estimation permits to characterize precisely the patient's health state, to choose the most rational approach to the treatment and to analyze its efficacy. PMID- 11482210 TI - [Correction of esophageal contracture in its cicatricial stenosis after chemical burn in children]. AB - Esophageal contracture occurs in presence of its cicatricial changes and is characterized by shortening of the organ, the hiatal hernia of diaphragm formation and reflux-oesophagitis, which supports inflammation in the oesophagus and prevents epithelization of the affection zone, causing further stenosing. In 11 children, aged from 1.5 to 6 yrs, with postburn cicatricial oesophageal stenosis the operative intervention was performed, consisting of oesophageal mobilization in mediastinum and its lengthening according to method of Collis with simultaneous fundoplication. Performance of the oesophageal contracture correction in children had permitted to prevent oesophageal inflammation, to restrict the stenosis progress, secured to do segmental oesophageal resection in 2 observations. PMID- 11482211 TI - [Aspects of clinical course and treatment of the gastroduodenal ulcer perforation in combination with hemorrhage]. AB - The results of treatment of 51 patients with gastroduodenal ulcer perforation (GDUP) in combination with hemorrhage were analyzed. Frequency of this complication in the gastroduodenal perforative ulcers structure had constituted 2.5%. The GDUP clinical variants in combination with hemorrhage were delineated depending on the time of the particular complication occurrence, the peculiarities of their clinical course were depicted. Necessity of an early radical operative intervention was substantiated as well as application of active surgical tactics while postoperative hemorrhage occurrence. Postoperative mortality was 29.4%. PMID- 11482212 TI - [Application of apparatus AKA-2 in the reconstructive-restorative surgery of the left half of colon and of rectum]. AB - Result of the AKA-2 apparatus application for the colorectal anastomosis formation in 129 patients after resection of left colonic half and rectum was estimated. After the operation all the patients survived. Insufficiency of the anastomotic edges conjunction have occurred in 6 (4.7%), cicatricial stricture- in 3 (2.3%) patients. Common postoperative complications were observed in 28 (21.8%) of the patients examined. PMID- 11482213 TI - [Chronic arterial insufficiency of lower extremities and pelvis: the ways of its treatment optimization]. AB - Results of application of preparation alprostan in 30 patients with chronic arterial insufficiency of lower extremities of III-IV stages were presented. Changes of regional blood flow and microcirculation of lower extremities under the influence of alprostan were studied. Positive result of treatment was achieved in more then 73% of patients. PMID- 11482214 TI - [Dose-dependent extravasal correction of relative insufficiency of the deep veins valves in varicose disease]. AB - For the 1989-1999 years period in the Donetsk centre of vascular surgery were operated 211 patients with varicose disease complicated by relative insufficiency of valves of deep veins. Of them 75 patients were operated using the method elaborated in the clinic, permitting to restore the valves function completely in 97.3% of observations. Application of conventional methods of extravasal correction of valve had secured positive result in 82.4% of observations. PMID- 11482215 TI - [Bacteriological investigation and treatment strategies of patients with diabetic foot complicated by purulent necrosis]. AB - Results of examination of 48 patients with diabetic foot (DF), in whom bacteriological investigation of tissues was conducted on various distance from inflammatory focus and in the blood serum (in 30 patients), were analyzed. The role of endogenous mechanism of infection and spread of microorganisms far away the purulent focus borders was established. The data obtained are permitting to perform rational treatment of patients with DF. PMID- 11482216 TI - [Treatment of purulent-inflammatory complications of the soft tissues wounds in elderly and senile patients]. AB - Experience of treatment of elderly and senile patients with purulent infection of the soft tissues wounds was summarized. For raising of the therapeutic effect of the local treatment the introduction of new combined medicinal forms, composition of which was scientifically substantiated according to the wound process pathogenesis with special attention to its phase and character of microflora (kind of microorganism and its sensitivity to antimicrobic agents), is perspective. PMID- 11482217 TI - [Application of ultrasonic investigation in diagnosis of "high" forms of an acute paraproctitis]. AB - Possibilities of ultrasonic investigation (USI) in diagnosis of "high" forms of an acute paraproctitis, ischiorectal and pelviorectal one, were studied. Of 20 patients examined, in 14 the diagnosis difficulties were noted, for correction of which the USI method was applied. The presence of purulent formations in pararectal cellular tissue was confirmed, their localization and spread were determined, permitting to substantiate the rational method of surgical intervention choice. PMID- 11482218 TI - [Fundamentals of anesthesiology-resuscitation. Part I. Methodology of the concept]. AB - The patient's reconvalescence, who fall ill suddenly or is injured or operated on, is impossible without the organism biological integrity (OBI) preservation or restoration and this constitutes the essence of the anesthesiology-reanimatology (AR) subject. The OBI is regarded as a process of necessary realization for the morphostructural balance preservation and of homeokinetic interaction of biological and energodynamical potential of organism. The OBI weak chains are determined, which are responsible for the course and the progression rate of bioenergetic insufficiency, acidotic alteration and the morphostructural balance disorder. The technology is adduced for the injuries elimination using the system of the local integrity restoration and the restoration OBI system. The AR application would raise the clinician's methodological armamentarium and promote to improve the result of treatment of patients in critical state. PMID- 11482219 TI - [Antibacterial therapy of pancreonecrosis]. PMID- 11482220 TI - [The complications in surgical treatment of gastric cancer in elderly and senile patients]. AB - Causes of the perioperative complications occurrence in 736 elderly and senile patients, operated on for gastric cancer, were studied. It was established that the complications frequency and mortality were depended on the concomitant diseases presence, on the main disease complications, duration and volume of operation as well, but not from their age. The operation performance method and anesthesiological support perfection would permit to widen indications for the radical surgical intervention performance and raise the treatment of patients efficacy. PMID- 11482221 TI - [Radical combined colonic resection for cancer recurrence]. AB - Experience of reoperative radical combined colonic resection for colonic cancer in 9 patients was presented. There were no complications, nor mortality. The preoperative radiation therapy and chemotherapy conduction had permitted to create an optimal conditions for the surgical intervention performance, which promoted the patients life span increase and improved its quality significantly. PMID- 11482222 TI - [The impact of hemodilution and low-molecular-weight fraxiparin on volumetric indexes and frequency of postoperative complications in surgical treatment of patients with oncological diseases]. AB - The influence of moderate hemodilution (HD) and of low-molecular-weight heparin (LMWH) fraxiparine on volumetric indexes (VI) and postoperative complications frequency in 38 patients with cancer recti, to whom intraoperative blood transfusion was not performed, was estimated. Control group consisted of 45 patients, in whom the preserved blood was used for restoration of the blood loss. While application of moderate HD and LMWH the VI had lowered in less extent than while performance of the erythrocytes mass transfusion, the postoperative complications frequency was also lower. PMID- 11482223 TI - [Simultaneous two-sided reconstruction of mammary gland in a woman with postmastectomy defect]. PMID- 11482224 TI - [Meckel's diverticulum in emergency surgical practice]. PMID- 11482225 TI - [Method of the blood evacuation from pleural and abdominal cavities in the presence of adhesions]. PMID- 11482226 TI - [Observation of the abdominal sac tumor]. PMID- 11482227 TI - [Observation of esophageal mycosis after gastric cancer surgery]. PMID- 11482228 TI - [Foreign body-related acute ileus]. PMID- 11482229 TI - [The change of gonadal activity in patients surgically treated for myasthenia gravis]. PMID- 11482230 TI - Public agency collaboration: implementation of the behavioral risk factor survey in adjacent Texas communities. AB - This article describes a collaborative community survey project between two public agencies serving adjacent urban communities. The main purpose of the collaboration was to establish comparable baseline health risk data for each community. To meet this goal, the agencies jointly administered the Centers for Disease Control and Prevention Behavioral Risk Factor Surveillance System questionnaire in their communities. The collaborative process was manageable and conserved both time and resources. This assessment project exemplifies a collaboration that both public and private healthcare entities can model to help ensure the health of the communities they serve. PMID- 11482231 TI - Sheila Leatherman on healthcare policy and evaluation and international quality agendas. Interview by Patti Klingel. AB - Sheila Leatherman has broad experience in health management, public policy, and academic research. She has held positions in federal and state healthcare agencies and served as CEO of a large HMO and as an executive of one of the largest managed care companies in the United States. Ms. Leatherman's time is currently devoted to healthcare research, both in the United States and the United Kingdom. In the United States, she is founder and chair of the Center for Health Care Policy Evaluation, an independent private-sector research institute, and adjunct professor in the Department of Health Policy and Management, School of Public Health at the University of North Carolina. In the United Kingdom, she is a senior adviser to The Nuffield Trust, which commissioned her to evaluate the National Health Service strategy for quality of care management (1997-1998 and 2002-2003), and in which she is involved in an ongoing policy and research program in quality evaluation. She is a Senior Associate at the Judge Institute of Management Studies and Distinguished Associate at Darwin College, both at Cambridge University. Her areas of research are quality of care, health policy, and managed care, and she has written more than 25 articles, which have been published in such prestigious journals as the Journal of the American Medical Association, Health Affairs, Medical Care, and the International Journal of Quality. PMID- 11482232 TI - Hospital nursing benchmarks: the California Nursing Outcomes Coalition project. AB - The California Nursing Outcomes Coalition (CalNOC) project is an initiative that has become the largest ongoing nursing quality measurement repository in the nation. Launched in 1996 by California nursing leaders concerned with trends in hospital care, CalNOC has created reliable quality benchmark data to define patient safety thresholds in California. This article describes CalNOC's effort, which aligns with the strategy of the National Quality Forum for measuring and reporting healthcare quality. By tracing the evolution of the CalNOC project and its future potential, we hope to encourage other grassroots efforts to build the database repositories needed for healthcare quality measurement in the 21st century. PMID- 11482233 TI - Children's hospitals collaborate to improve quality of care. AB - The Child Health Accountability Initiative brought together 35 chief executive officers, physicians, and researchers representing 12 children's hospitals and charged them with what most would think is a lofty goal: to improve health outcomes for America's children. After a rigorous and often highly debated selection process, two data-driven performance improvement projects were rapidly set in motion and implemented in each of the hospitals, with significant results- all within the first year. The initiative's breadth, scope, and speed may serve as a model for other healthcare collaboratives. PMID- 11482234 TI - A guide to using performance measurement systems for continuous improvement. AB - The Joint Commission on Accreditation of Healthcare Organizations requires accredited organizations to use a performance measurement system that meets its inclusion requirements to satisfy performance outcome and measurement expectations. The system, known as the ORYX initiative, is used for both internal performance control and external performance comparisons. This article outlines a three-step approach to using a performance measurement system based on the philosophy of continuous improvement and the methods of statistical process control (SPC). SPC, the methodology recommended by the Joint Commission, can be applied to the analysis of many quality measures and can be implemented with Microsoft Excel software. PMID- 11482235 TI - A Symposium on E-healthcare Strategies for Physicians, Hospitals, and Integrated Delivery Systems. PMID- 11482236 TI - Medical privacy: a top-of-mind issue with patients, providers, payers, and legislators. PMID- 11482237 TI - Developing performance measures for sedation and analgesia: the approach of the Quality Indicator Project. AB - This article describes the development of a set of measures focused on use of and complications following sedation and analgesia. This group of measures is another step in a 15-year quest of a group of hospitals and other healthcare providers to identify ways to better monitor and understand performance--and thereby improve their ability to identify opportunities for improvement. The article describes in detail the practical steps taken to develop, test, and implement the measures, as well as some of the fundamental conceptual issues associated with cost and benefits of performance measurement. Finally, it examines the pilot-test experience for the measures--both the individual hospitals' efforts to implement data collection and the aggregate data that resulted from the pilot test. PMID- 11482238 TI - Interview with Peter Butler, president and CEO, Methodist Health Care System. Interview by Kyle L. Grazier. PMID- 11482239 TI - The voice of the customer: is anyone listening? PMID- 11482240 TI - Magnet hospitals use culture, not wages, to solve nursing shortage. PMID- 11482241 TI - Ensuring management excellence in the healthcare system. AB - We believe that if we improve healthcare management, we will improve quality, reduce cost, and open opportunities to expand access. The present support for healthcare management, however, needs improvement in every aspect. Beginning with attracting young people, progressing through entry education, continuing education, and the special preparation for senior management, what we need overall is a program of continuous improvement. We need specific goals, measures of how well we achieve those goals, and accountability for that achievement. We need programs that attract and reward excellence and that make mediocrity untenable. The purpose of this article is to identify and expand the elements of such a program. PMID- 11482242 TI - Managing strategic outsourcing in the healthcare industry. AB - Hospitals and healthcare systems are facing increased financial difficulties because of the Balanced Budget Act of 1997 and managed care. As a result, healthcare executives face the challenge of reducing costs while maintaining quality patient care. One of the strategic tools healthcare executives use to meet this challenge is outsourcing. Even though outsourcing has many benefits, outsourcing will fail if not managed successfully. Senior executives must choose outsourcing managers who have the necessary leadership capabilities. Managing outsourcing requires an understanding of outsourcing strategy, the benefits and risks of outsourcing, the evaluation process, and the methods to managing strategically. With appropriate management, strategic outsourcing should provide healthcare executives with a viable strategy for controlling costs and maintaining quality patient care. PMID- 11482243 TI - The adoption of complementary and alternative medicine by hospitals: a framework for decision making. AB - As a result of increased consumer awareness, personal preference, and limitations of conventional medicine, many individuals are turning to complementary and alternative medicine (CAM). In response to this movement, many community hospitals are striving to be innovative providers. Society is leaning toward a more comprehensive style of healing that incorporates all aspects of wellness. During the last three decades, the public has increasingly used CAM. Arnold (1999) cited a study published in the Journal of the American Medical Association that reported that 39 million people sought either advice or treatment from a CAM provider and 42 percent of Americans used some form of alternative therapy. With the population becoming increasingly educated, aware, and proactive about wellness, many Americans see CAM as an effective alternative to traditional medicine. Healthcare organizations have responded, although slowly, to this trend, as new alternative medicine clinics, hospital departments, and research centers emerge throughout the United States. Although alternative medical practices are being used by an increasing number of people, there is still limited understanding of what CAM includes and how it influences health services organizations. Understanding this new market and its implementation in the healthcare setting is of interest to healthcare administrators. This article defines CAM, discusses its rising popularity, identifies its adoption in today's hospitals, and depicts barriers to its implementation. Finally, an analytical framework developed by the author is used to suggest factors for administrators to consider in CAM implementation in their organizations. PMID- 11482244 TI - Clinical service lines in integrated delivery systems: an initial framework and exploration. AB - The increasing pressures on integrated healthcare delivery systems (IDSs) to provide coordinated and cost-effective care focuses attention on the question of how to best integrate across multiple sites of care. One increasingly common approach to this issue is the development of clinical service lines that integrate specific bundles of services across the operating units of a system. This article presents a conceptual model of service lines and reports results from a descriptive investigation of service line development among members of the Industry Advisory Board--a research consortium comprising IDSs. The experiences of these IDSs (1) provide valuable insights into the range of organizational arrangements and implementation issues that are associated with service line management in healthcare systems and (2) suggest aspects of service line management worthy of further inquiry. PMID- 11482245 TI - Physician recruitment strategies for a rural hospital. PMID- 11482246 TI - Predictive modeling care management program produces dramatic bottom line results. AB - Predictive modeling program identifies high-risk patients before complications lead to skyrocketing costs. It's a new-generation effort, but it has already proven its worth in a large population, producing savings of $1 million to $2 million per month. The effort begins with the development of a constantly evolving registry of high-risk patients. Specialized case management efforts are then focused on stabilizing individuals within this group. See how traditional interventions have been combined with new technology to deliver results. PMID- 11482247 TI - Use this report card to profile physicians on pharmacy use. AB - With the emphasis on reducing pharmacy costs and utilization, here's a simple profiling tool that can reveal prescribing patterns at a glance. The objective is to accomplish those goals while keeping quality measures foremost. PMID- 11482248 TI - Breakthrough found in treatment of severe sepsis. AB - By late summer, hospitalists may have in hand a medication to help them fight sepsis, which causes 1,400 deaths daily throughout the world. PMID- 11482249 TI - UCLA's coronary artery disease initiative begins at the hospital with medication, lifestyle interventions. AB - Begin cardiac rehab in the hospital. Re-think the conventional wisdom about when to start cardiac rehab therapies. Researchers at the University of California Los Angeles are beginning right after heart surgery with education and medication regimens that used to wait until a patient returned to home. PMID- 11482250 TI - Expedient treatment key to fighting meningococcal disease. PMID- 11482251 TI - Length of stay in myocardial infarction. AB - OBJECTIVE: We evaluated the association between length of hospital stay (LOS) and clinical factors, treatment intensity, and use of percutaneous coronary revascularization from 1988 to 1997. BACKGROUND: Multiple factors contribute to the observed reduction in LOS for patients with myocardial infarction. METHODS: We studied a series of 849 consecutive patients admitted with acute myocardial infarction to the Mayo Clinic Coronary Care Unit within three time periods: period I (1988-1990), period II (1991-1993), and period III (1994-1997). RESULTS: Median LOS decreased significantly between 1988 and 1997 (9 days to 5 days, 36% reduction, p < 0.0001), with significant reductions (p < 0.001) associated with certain therapies: primary reperfusion (6 days vs 7 days), b-blockers (6 days vs 8 days), and aspirin (6 days vs 8 days). Hospitalizations were lengthened by coronary artery bypass grafting (12 vs 6 days) and by serious complications (10 vs 6 days). The era of the admission (period I vs II vs III) is a significant, powerful predictor of LOS, even after adjustment for other key variables. CONCLUSION: The 36% reduction in LOS for acute myocardial infarction between 1988 and 1997 is related both to therapeutic modalities and temporal trends. Further study is needed to clarify whether the trend for decreasing LOS persists and influences outcome and health care quality variables. PMID- 11482252 TI - Hospitals and hospitalists can reach across the "quality chasm" by restructuring inpatient care. PMID- 11482253 TI - The missing link: management and employee retention. PMID- 11482254 TI - Six providers speak out about managed care, reimbursement, and real life. Roundtable discussion. PMID- 11482255 TI - An inventory of patients. Workflow concepts from manufacturing can be applied in health care. PMID- 11482256 TI - The "oh, no!" syndrome. Are health care institutions unwilling to deal with overcrowding and patient flow issues in the ED? PMID- 11482257 TI - FDA scrutiny and pediatric clinical investigations. PMID- 11482258 TI - Do you have to accept this patient? What intensivists and hospitalists need to know about EMTALA. PMID- 11482259 TI - Conquering the technology summit. Online purchasing activity continues upward climb. PMID- 11482260 TI - An independent's certified value. Third-party credentialing ensures unbiased results. PMID- 11482261 TI - Finding sterile technology. PMID- 11482262 TI - Hip standards. Strides to cut the rising cost of implants. PMID- 11482263 TI - An overview of health research funding in Canada. PMID- 11482264 TI - Optimizing the relationship between the academic health centre and the community hospital. PMID- 11482265 TI - Putting patients first: the role of nursing caring. PMID- 11482266 TI - Who knows best: the patient or the provider? A nursing perspective. AB - Who knows best: the patient or the provider? My opinion, based on a review of the literature and practical experience, is that the patient, the provider and the system each offer a unique perspective that we can draw upon in ensuring quality care across the continuum. Nurses have a unique body of knowledge and skill that they bring to each interaction with the patient. They must have an awareness of the patient's and the system's expectations and interact and negotiate realistic expectations for each. The maintenance of balanced expectations and the measurement of effectiveness will continue to be a challenge. However, patients should be involved in and direct aspects of their care and feel satisfied with the process. Ultimately, nurses and the hospitals in which they work are responsible for providing effective and satisfying care. I would like to end by reinforcing the importance of the patient's voice in the provision of compassionate care. It is disheartening to read about patients' experiences of "discompassionate healthcare" (Holloway 1999). Yet I am reassured when I read or experience examples of compassionate and effective care such as that outlined by Valerie McDonald. (Hospital Quarterly Winter 1999/2000) Ms. McDonald, a former social worker and adult educator, is the mother of three daughters, one who had Burkitt's lymphoma diagnosed in 1994 and a second who had acute lymphostatic leukemia diagnosed in 1997 and who died recently in 1999. McDonald provided a wonderful perspective about her hospital experiences--the good and the bad. There would be no denying from this report that patients know the key qualities necessary for effective and compassionate care and that this mother recognized the energy and time it took to provide this care. "I hope," she states, "as the dust settles from restructuring and cutbacks that hospital staff will still have the time, energy and flexibility to practice the art of healing as they did with my children" (p. 24). I too reinforce that we must ensure nurses (and others) have the resources, the flexibility within their roles and the knowledge and skill to practise both the art and science of nursing. PMID- 11482267 TI - Best practices in the care of elderly persons in hospital. PMID- 11482268 TI - Supportive care for cancer patients. PMID- 11482269 TI - Use of mid-level indicators in determining organizational performance. PMID- 11482270 TI - The spin doctor is in. PMID- 11482271 TI - The future of the Internet in healthcare: a five-year forecast. PMID- 11482272 TI - Canadians speak out for home care. PMID- 11482273 TI - Thank God for rich women with mustaches. PMID- 11482274 TI - For the wounds that will never heal. PMID- 11482275 TI - Mohs micrographic surgery: an overview. PMID- 11482276 TI - From vitamin to Vesanoid: systemic retinoids for the new millennium. AB - Retinoids are a fascinating class of compounds that exert control over cellular function from the time of conception to death. They play a critical role in such vital processes as fetal morphogenesis, cellular differentiation and apoptosis. Over the years synthetic retinoids have provided dermatologists with a spectrum of medications that have profound therapeutic effects on a variety of recalcitrant skin disorders. Moreover, retinoids are an expanding component of the treatment arsenal against hematologic and solid malignancies. Retinoids are poised to offer exciting new therapeutic options in the field of endocrinology for the treatment of diabetes and lipid disorders. Researchers and clinicians are only beginning to unveil the therapeutic potential of this class of medications. The development of new retinoid compounds targeting specific receptors promises a wealth of new therapies for the new millennium. PMID- 11482277 TI - The value of a diagnostic punch biopsy. PMID- 11482278 TI - Epidemiology, etiology, and control of melanoma. PMID- 11482279 TI - Ultraviolet radiation and sunscreens. PMID- 11482280 TI - Images in medicine. Ganglioneuroma. PMID- 11482281 TI - Generic drug substitution. PMID- 11482282 TI - Problems in the control of melanoma of skin, Rhode Island, 1987-1998. PMID- 11482283 TI - Race, ethnicity, and health: a new data policy. PMID- 11482284 TI - Cataracts and hernias: aspects of surgical practice in the fourteenth century. PMID- 11482285 TI - Civil liberties and public good: detention of tuberculous patients and the Public Health Act 1984. PMID- 11482286 TI - New World materia medica in Spanish renaissance medicine: from scholarly reception to practical impact. PMID- 11482287 TI - The physician in ancient Israel: his status and function. PMID- 11482288 TI - "A most hideous object": John Davies (1796-1872) and plastic surgery. PMID- 11482289 TI - The role of the divine proportion in the esthetic improvement of patients undergoing combined orthodontic/orthognathic surgical treatment. AB - This study was designed to investigate the changes in a number of facial proportions as a result of combined orthodontic/orthognathic surgical treatment. According to some authors, in beautiful faces, the values of the proportions measured are likely to approximate the divine proportion (1.618:1). The hypothesis for this study was that, as a result of treatment, the faces of patients in the sample would be more esthetic and therefore the measured proportions would be closer to the divine proportion than they were before treatment. Forty-six patients were included in the sample. Pre- and posttreatment photographs were each given a score (out of a possible 100) by 12 judges using a visual analog scale. Judges were shown frontal and profile views simultaneously. Ten ratios were measured from pre- and posttreatment lateral cephalograms, and 11 were measured from the frontal photos. Pearson's correlation coefficient was used to determine the correlation between changes in esthetic rating and changes in the measured proportions. No correlations were found between changes in esthetic ratings and changes in the proportions. While most subjects were considered more esthetic after treatment than before, the proportions were equally likely to move away from or toward the divine proportion. For this reason, if it is to be used as an aid to orthodontic/orthognathic treatment planning, the divine proportion should perhaps be used along with other methods of anteroposterior and vertical cephalometric and facial assessment. PMID- 11482290 TI - Stability of sagittal split ramus osteotomy used to correct Class III malocclusion: review of the literature. AB - Setback of the mandible to correct mandibular prognathism is a well-known procedure. The 2 most frequently used techniques are the intraoral vertical ramus osteotomy (IVRO) and the sagittal split ramus osteotomy (SSRO). Although SSRO has been performed for many years, few data exist concerning long-term skeletal stability, and different hypotheses have been suggested to explain potential relapse. The literature published between 1985 and the present concerning this procedure was reviewed, and the authors analyze and discuss skeletal stability and factors contributing to relapse. PMID- 11482291 TI - Postoperative migration at the individual osteotomy site following sagittal split ramus osteotomy: a stereometric radiographic study. AB - The aim of the present study was to evaluate whether migration occurs at individual osteotomy sites following sagittal split osteotomy of the mandible stabilized with rigid internal fixation, and if it occurs, how long it lasts. In 10 consecutive patients operated with bilateral sagittal split osteotomies, postoperative migration, defined as displacement of the proximal segments in relation to the distal over time, was studied 3-dimensionally by means of stereometric radiography. Follow-up was performed with stereometric radiographs obtained at intervals from 2 days until 1 year after surgery. During the 1-year observation period, migration at one or both of the osteotomy sites was found at some stage in all of the patients and in as many as 4 patients during the final 6 months. It is concluded that rigid internal fixation, as performed in the present study, does not prevent postoperative migration at the osteotomy. Furthermore, stable bone union at the osteotomy site appears to be a slower process than previously expected, thus emphasizing the importance of follow-up procedures to prevent relapse. PMID- 11482292 TI - Complications following orthognathic surgery that required early surgical intervention: fifteen years' experience. AB - The aim of this study was to assess complications following various orthognathic surgical procedures that required early surgical intervention. This study was carried out on 821 patients who had undergone surgical treatment for correction of dentofacial deformities between 1985 and 2000. Only patients who required a second procedure to deal with immediate or early postoperative complications (i.e., those occurring within 4 weeks of surgery) were investigated in this study. Twelve patients underwent a second surgical procedure; 9 had undergone conventional osteotomy surgery, and 3 had undergone distraction osteogenesis. Three Le Fort I cases had to be further impacted and repositioned, and 4 vertical subsigmoid osteotomies had to be reexplored. The details of the complications are presented, and possible methods by which these problems could be reduced and/or prevented are discussed. PMID- 11482293 TI - Temporomandibular dysfunction in 1,516 patients before and after orthognathic surgery. AB - Functional disturbances, together with esthetic considerations, are important reasons for patients to seek orthognathic surgical treatment. Functional disorders may include signs and symptoms of temporomandibular disorders (TMD), such as joint pain, chewing problems, joint noises, headaches, etc. This paper reports on TMD before and after orthognathic surgery in 1,516 patients. It is based upon the patients' own evaluations as recorded 2 years after surgery. Preoperatively 43% and postoperatively 28% of the patients reported subjective symptoms of TMD. This difference indicates an overall beneficial effect of orthognathic surgery on TMD signs and symptoms. Patients with mandibular retrognathia did not improve as much as patients with mandibular prognathia. Sagittal ramus osteotomy was less effective than vertical ramus osteotomy in relieving TMD symptoms when performed on similar diagnoses. PMID- 11482294 TI - Effect of orthognathic surgery on the temporomandibular joint in patients with anterior open bite. AB - This study examined the prevalence of temporomandibular joint (TMJ) signs and symptoms in patients with anterior open bite. The influence of orthognathic surgery on the TMJ in these patients and the interaction of occlusal and psychologic variables on the presence and/or persistence of pain was studied. A retrospective survey of 83 patients with an anterior open bite who underwent orthognathic surgery was carried out. Records were examined for the prevalence of abnormal TMJ signs and symptoms, including pain. A survey was mailed to these patients that consisted of: (1) the TMJ Scale, (2) the Symptom Checklist 90 (SCL90), (3) the Spielberger State-Trait Anxiety Inventory (STAI), and (4) a visual analog scale on which patients indicated their degree of satisfaction with the procedure. Thirty-seven (42%) patients responded to the survey, and 13 (15%) also attended a clinical and radiographic examination. Multiple regression analysis was used for statistical analysis of the factors contributing to the presence and/or persistence of pain. In the preoperative group, the prevalence of pain was 32%, dysfunction 40%, and limitation of opening 7%. Age and gender were significantly associated with the presence of pain. The overall prevalence of abnormal TMJ signs and symptoms was not significantly different after orthognathic surgery. An abnormal psychologic profile was the most significant factor associated with the presence and/or persistence of pain. It is concluded that that the prevalence of temporomandibular disorders in anterior open bite patients increases with age, is significantly higher in females, and is not influenced by other occlusal variables. Furthermore, orthognathic surgery does not significantly influence temporomandibular disorders in patients with anterior open bite. Female patients, particularly those with an abnormal psychologic profile, are at a higher risk of persistent postoperative TMJ pain. PMID- 11482295 TI - Effects of a computerized treatment simulation on patient expectations for orthognathic surgery. AB - One hundred forty-six patients, ranging in age from 15 to 50 and presenting with a moderate to severe dentofacial disharmony requiring orthodontics and orthognathic surgery, were randomly assigned to 2 preparation strategy groups: standard presurgical consultation with or without a computerized treatment simulation presentation. The demographic profiles of the 2 groups were similar. Viewing a treatment simulation did not have a significant effect on the anticipation of social/interpersonal or general health problems in the first month after surgery. These 2 areas of concern were significantly related to psychological well-being. Psychologically distressed patients, whether or not they saw a simulation, expected significantly more problems in social/interpersonal relations and in general health during the first month after surgery. A treatment simulation presentation did affect patients' overall expectations of problems in the first month after surgery and their concerns about symptom recovery. However, the impact of the presentation was related to the patient's psychological well-being. In the standard presurgical consultation group, the average anticipated level of overall problems and discomfort during recovery was significantly higher for patients who reported elevated psychological distress than for those who did not. In the treatment simulation group, the average level of concern was similar for those patients who reported distress and those who did not. Preparation strategy was not significantly related to the long-term expectation of improvement after treatment. Long-term expectation of treatment improvement was related to psychological distress and gender. Men tended to report similar expectations regardless of psychological well-being, while women who were distressed anticipated significantly more improvement overall, in self-image, and in general health after treatment than women who were not distressed. PMID- 11482296 TI - Patient satisfaction following orthognathic surgical correction of skeletal Class III malocclusion. AB - The objectives of this study were to evaluate the relationship between preoperative psychologic status and attitude and postsurgical adjustment and experience of the surgical treatment. Questionnaires were sent to 140 consecutive Chinese patients with skeletal Class III malocclusion who had been treated with a combined orthodontic-surgical approach. The results showed the following: (1) immediately after the surgery 44% of patients had more pain, 57% had more numbness, and 73% had more swelling than expected; (2) most patients underwent noted marked changes in facial appearance (96%) and dental appearance (91%); (3) chewing ability improved in 71% of patients; (4) half of those with temporomandibular joint problems preoperatively experienced improvement; (5) personality and lifestyle were affected positively in about 50% of patients; (6) satisfaction with the treatment increased with time: 87% at 6 months and 92% at 24 months; and (7) a few patients (8%) regretted having undergone surgery, mainly because facial changes were not apparent. The vast majority of the present Chinese skeletal Class III patients were satisfied with the overall outcome of the continued orthodontic-surgical treatment. PMID- 11482297 TI - Commentary on Greene and Ablon: What does the MTA study tell us about effective psychosocial treatment for ADHD? PMID- 11482298 TI - International Society of Nephrology: a forty year history. 1960-2000. PMID- 11482299 TI - Does hyaluronate prevent postoperative retro-sternal adhesions in coronary surgery? --Preliminary results. PMID- 11482300 TI - Thoracoscopic sympathectomy for craniofacial hyperhidrosis. PMID- 11482301 TI - Initial experience with a new communication tool: the 'Digital Interactive Poster Presentation'. PMID- 11482302 TI - C1q does not complement the removal of apoptotic keratinocytes in vivo. PMID- 11482303 TI - Sensing and killing bacteria in skin: innate immune defense system: good and bad news. PMID- 11482304 TI - Blood pressure and symptoms of depression and anxiety: a prospective study. AB - This study investigated whether symptoms of depression and anxiety were related to the development of elevated blood pressure in initially normotensive adults. The study's hypothesis was addressed with an existing set of prospective data gathered from an age-, sex-, and weight-stratified sample of 508 adults. Four years of follow-up data were analyzed both with logistic analysis, which used hypertension (blood pressure > or =140 mm Hg systolic or 90 mm Hg diastolic) as the dependent variable, and with multiple regression analysis, which used change in blood pressure as the dependent variable. Five physical risk factors for hypertension (age, sex, baseline body mass index, family history of hypertension, and baseline blood pressure levels) were controlled for in the regression analyses. Use of antidepressant/antianxiety and antihypertensive medications were controlled for in the study. Of the 433 normotensive participants who were eligible for our study, 15% had missing data in the logistic regression analysis focusing on depression (n = 371); similarly, 15% of the eligible sample had missing data in the logistic regression using anxiety as the psychological variable of interest (n = 370). Both logistic regression analyses showed no significant relationship for either depression or anxiety in the development of hypertension. The multiple regression analyses (n = 369 for the depression analysis; n = 361 for the anxiety analysis) similarly showed no relationship between either depression or anxiety in changes in blood pressure during the 4 year follow-up. Thus, our results do not support the role of depressive or anxiety symptoms in the development of hypertension in our sample of initially normotensive adults. PMID- 11482305 TI - [The future of rehabilitation--orientation for services of the Federal Society of Rehabilitation beginning 2001]. PMID- 11482306 TI - Less specific myositis autoantibodies? PMID- 11482307 TI - Usefulness of the HAQ in the clinic. PMID- 11482308 TI - Multinodular goitre with giant cell vasculitis of thyroid arteries in a woman with temporal arteritis. PMID- 11482309 TI - Giant cell arteritis associated with demyelinating polyradiculoneuropathy. PMID- 11482310 TI - Oral steroid in the treatment of carpal tunnel syndrome. PMID- 11482311 TI - Parvovirus B19 infection in Behcet's disease. PMID- 11482312 TI - Idiopathic dilatation of the pulmonary artery in a patient with dermatomyositis complicated by interstitial pneumonitis. PMID- 11482313 TI - Arthritis associated with non-alcoholic steatohepatitis. PMID- 11482314 TI - The use of 2D and 3D displays for shape-understanding versus relative-position tasks. AB - Research on when and how to use three-dimensional (3D) perspective views on flat screens for operational tasks such as air traffic control is complex. We propose a functional distinction between tasks: those that require shape understanding versus those that require precise judgments of relative position. The distortions inherent in 3D displays hamper judging relative positions, whereas the integration of dimensions in 3D displays facilitates shape understanding. We confirmed these hypotheses with two initial experiments involving simple block shapes. The shape-understanding tasks were identification or mental rotation. The relative-position tasks were locating shadows and determining directions and distances between objects. We then extended the results to four experiments involving complex natural terrain. We compare our distinction with the integral/separable task distinction of Haskel and Wickens (1993). Applications for this research include displays for air traffic control, geoplots for military command and control, and potentially, any display of 3D information. PMID- 11482315 TI - [Preventability of adverse effects in a medical emergency service]. AB - The aim of this prospective pharmacovigilance study was to assess the incidence and the preventability of adverse drug reactions (ADRs) leading to hospital admissions. All patients admitted to the Toulouse University Hospital through the medical admission ward during four non-consecutive weeks were included in the study. Characteristics of patients admitted for a suspected ADR (cases) were compared with those admitted for other reasons (controls). All cases were reviewed by both a pharmacologist and an emergency medicine specialist. Among a total of 671 admissions, 44 ADRs were identified. The incidence of hospital admissions for ADRs was 6.1 per 100 admissions [4.4-8.3]. Cases were exposed to a higher number of drugs (3.6 vs. 1.7; p < 0.001). Some classes of drugs were significantly associated with a higher risk of ADRs: antineoplastic, anti infectious or musculoskeletal drugs. Haematological, metabolic and electrolytic, liver/gastrointestinal or cutaneous disorders were causes of admissions significantly more frequently related to ADRs. Using Imbs's preventability scale, 65 per cent of ADRs were categorized as 'definitely unpreventable', 26 per cent 'potentially preventable' and 9 per cent 'definitely preventable'. These results underline the frequency of ADRs leading to hospitalization, with 35 per cent which are more than likely preventable. Further studies are needed to validate this preventability scale in order to obtain an easier, more reliable and more reproducible tool. PMID- 11482316 TI - [Experimental studies of the value of SPIO for MRI of bone marrow before and after whole body irradiation]. AB - PURPOSE: Evaluation of the value of superparamagnetic iron oxides (SPIO; Endorem) for MRI-derived quantifications of the permeability of the blood-bone marrow barrier and the phagocytic activity of reticuloendothelial system (RES) bone marrow cells before and after TBI. METHODS: 12 New Zealand white rabbits underwent MRI of the lumbar spine and os sacrum using T1-weighted spinecho (SE) and T2-weighted Turbo-SE (TSE) sequences before and after injection of SPIO (Endorem). Four animals each were examined without irradiation, after 4 Gy total body irradiation (TBI), and after 12 Gy TBI. Changes in bone marrow signal intensities (SI) after contrast agent injection were quantified as delta SI(%) = SIpost-SIpre)/SIpre) x 100% and these data were correlated with bone marrow histopathology. RESULTS: Histopathology of the bone marrow revealed a radiation induced decline of all hematopoetic cell lines. SPIO were phagocytosed by bone marrow RES cells and caused a significant bone marrow signal decline on postcontrast T2-weighted images (p < 0.05). delta SI(%) data for T2-weighted images were significantly higher for the irradiated bone marrow as compared to non-irradiated controls (p < 0.05). Dynamic T1-weighted images directly after contrast medium injection were not able to characterize the permeability of the blood-bone marrow barrier. CONCLUSION: Hematopoetic bone marrow can be labelled with SPIO. Irradiation does not impair the phagocytic activity of bone marrow RES cells. However, the bone marrow enhancement with SPIO is smaller as compared to previous results obtained by our group with USPIO. PMID- 11482317 TI - [Prostatic benign hyperplasia]. AB - After reviewing the clinical, diagnostic and therapeutic attitude related to Benign Prostatic Hyperplasia, the authors present the guidelines for Generalists, in terms of practical handling and when and how to refer their patients to the Urology consultation. Based on the criteria presented, Generalists must refer their patients to the Urology consultation in the presence of mild or severe symptoms, ages below 50 years, diabetes or neurological disturbances, disturbed voiding, palpable bladder, palpable nodule or a diffusely hardened or asymmetric gland revealed by digital rectal examination, hematuria, urinary tract infection, renal impairment or PSA above 4 nglml in patients aged below 70 years. PMID- 11482318 TI - Increased prevalence of CMV gB3 in marrow of patients with aplastic anemia. PMID- 11482319 TI - Treatment of extensive chronic sclerodermatous graft-versus-host disease with high-dose immunosuppressive therapy and CD34+ autologous stem cell rescue. PMID- 11482320 TI - A comparison of polyethylene wear in hips with cobalt-chrome or zirconia heads. A prospective, randomised study. AB - We have reviewed 70 patients with bilateral simultaneous total hip arthroplasties to determine the rate of failure and to compare polyethylene wear and osteolysis between an implant with a cobalt-chrome head and Hylamer liner with that of a zirconia head and Hylamer liner. The mean thickness of the polyethylene liner was 11.0 mm (8.8 to 12.2) in the hip with a zirconia head and 10.7 mm (8.8 to 12.2) in that with a cobalt-chrome head. At follow-up at 6.4 years no acetabular or femoral component had been revised for aseptic loosening and no acetabular or femoral component was loose according to radiological criteria in both the cemented and cementless groups. The mean rate of linear wear and annual wear rate were highest in the 22 mm zirconia femoral head (1.25 mm (SD 1.05) and 0.21 mm (SD 0.18), respectively) and lowest in the 22 mm cobalt-chrome femoral head (0.70 mm (SD 0.39) and 0.12 mm (SD 0.07), respectively). The mean volumetric wear was highest in the 28 mm zirconia femoral head (730.79 mm ) and lowest in the 22 mm cobalt-chrome femoral head (264.67 mm3), but if the results were compared by size of the femoral head and type of material there was no statistical difference (p > 0.05). Sequential measurements of annual wear showed that the zirconia femoral head had a relatively higher rate of penetration than the cobalt-chrome head over the first three years; thereafter the rate of wear was reduced and compared favourably with that of cobalt-chrome heads. There was a statistically significant relationship between the wear of the polyethylene liner and the age of the patient, male gender and the degree of abduction angle of the cup, but not diagnosis, weight, hip score, range of movement, or amount of anteversion. Osteolysis was identified on both sides of the acetabulum in six patients (9%). Of 12 hips with acetabular osteolysis, six had a 28 mm cobalt-chrome femoral head and the remaining six a 28 mm zirconia head. Osteolysis was observed in zones 1A and 7A of the femur in two hips (3%) with a 28 mm zirconia head (cemented hip) and in four (6%) with a 28 mm cobalt-chrome femoral head (cementless hip). Our findings suggest that although the performance of a zirconia femoral head with a Hylamer liner was not statistically different from that of a cobalt-chrome femoral head and Hylamer liner, there was a trend for the zirconia head to be worse than the cobalt-chrome femoral head. PMID- 11482321 TI - Alternatives to xenotransplantation 1--increased human cadaveric donation. PMID- 11482322 TI - Council of Europe Working Party on Xenotransplantation: state-of-the-art report on xenotransplantation (2000). PMID- 11482323 TI - Patients, professionalism and politics: a new beginning. PMID- 11482324 TI - Soy formulas and the effects of isoflavones on the thyroid. PMID- 11482325 TI - Thirty years of universal home dialysis in Christchurch. AB - AIM: To review 30 years of universal home dialysis in a single dialysis unit. METHOD: Analysis for patients using home dialysis since 1969 of information from hospital visits, clinical case notes and demographic and survival data from the Australia and New Zealand Dialysis and Transplant Registry. RESULTS: Since 1969 treatment options at the Christchurch Nephrology Unit for patients with end-stage renal disease have been home haemodialysis (HD), renal transplantation and, since 1979, continuous ambulatory peritoneal dialysis (CAPD). No long-term, hospital based treatment has been offered. During this time 493 patients, aged 3-82 years, began treatment. The mean training time for home HD was 79 days (range 23-268) and for home CAPD 7 days (range 1-35). The mean HD treatment time was 7 hours x 3 per week (range 10-36 hours/week). Between 1980 and 1995, less than 5% of patients took antihypertensive drugs and 73% of those aged 18-65 years were in full or part-time employment. The mean age of patients commencing treatment increased from 41.8 years in the 1970s to 50.1 years in the 1990s. The median patient survival from 1970-1997 was 7.75 years on home HD and 2.1 years on home CAPD. Median survival on dialysis fell in the 1990's as more diabetics and older patients with comorbidity started treatment. CONCLUSIONS: Home HD allows good rehabilitation, long treatment times and good blood pressure control which may all contribute to the superior survival of home versus hospital HD. CAPD survival in Christchurch was worse than HD, but this is probably due to patient selection. A policy of universal home dialysis is still workable provided there are sufficient resources for training and support of patients in the community. PMID- 11482326 TI - An investigation into the incidence of toxoplasmosis in pregnancy in New Zealand. AB - AIM: To estimate the incidence of toxoplasmosis in pregnancy in New Zealand and consider whether there is a case for screening women in pregnancy. METHODS: The risk of maternal and fetal infection with toxoplasmosis was derived by first determining the rate of maternal seroconversion based on seroprevalence studies. The age-specific number of seroconversions in pregnancy was then estimated from the birth rate. Using reported fetal infection rates after primary maternal infection, the expected number of congenitally infected infants in one year was estimated. These incidences were compared with the number of recognised cases of toxoplasmosis infection in pregnancy and the actual number of positive IgM results at the Wellington Hospital laboratory. Using national births data, this incidence was extrapolated to estimate the number of expected cases in New Zealand. RESULTS: The annual seroconversion rate was 0.62% (95% confidence interval 0.39-0.86). On this basis, 164 primary maternal infections are expected annually with 66 fetuses being infected. Ten patients tested positive for IgM in Wellington, which averaged only one case per year being identified over the time examined in this study. CONCLUSIONS: Very few of the expected cases in pregnancy are diagnosed. Reporting rates were low when toxoplasmosis was a notifiable disease. Other means of improving detection, reporting and the avoidance of infection are discussed. More information is required before screening can be recommended in New Zealand. PMID- 11482327 TI - Emerging clinical governance: developments in independent practitioner associations in New Zealand. AB - AIMS: To document and analyse the development of independent practitioner associations and similar groups in New Zealand. METHODS: A questionnaire was sent to the 30 independent practitioner associations in August 1998 and followed-up by a number of reminders. RESULTS: The 28 respondents (93%) represent 97% coverage of the estimated membership of independent practitioner associations and similar groups. Membership of the 28 responding organisations ranged between seven and 340, with an average of 74 members and a total of 132 employed staff. Twenty-one had appointed a chief executive officer or general manager. The respondents' most important goals were "achieving better health outcomes for patients" and "making better use of primary care resources". They reported almost total implementation of computerised age/sex registers in their practices. There was strong support for independent practitioner associations to manage the clinical activity of members, to move from historical to equitable, needs-based funding and for formal patient enrolment. The majority of respondents supported integrated and capitated primary care budgets but few supported capitated budgets for separate general medical services, laboratory and pharmaceutical services. Important recent initiatives include a wide range of integration projects and increasing involvement of local communities. CONCLUSION: Independent practitioner associations have made significant progress in increasing membership levels, in establishing a framework for managing clinical activity of members and in developing their infrastructure, including information systems. They have established a wide range of new relationships within primary care, with their communities and with primary and secondary care providers. In managing increasing amounts of public money to achieve public goals, these groups may be developing a new model of clinical governance, which could be of international importance. PMID- 11482328 TI - Echocardiographic evaluation of stroke. PMID- 11482329 TI - Asthma in New Zealand: myths and realities. AB - Asthma is a major health problem in New Zealand, generating large costs to the health services and industry. However, there are several myths concerning asthma in New Zealand. These are that it is more common and more severe in New Zealand than in the rest of the world, that it is more common in rural areas and that asthma mortality in New Zealand is the highest in the world. Using recently published data, it is now possible to show that none of these "facts" is true. In fact, asthma in New Zealand appears to have a prevalence and severity similar to those of the other major English-speaking countries and in this respect can be thought of as being first equal in the world. The relevance of correcting these myths is that they could influence future research. Previous research into the causes of asthma has sometimes focused on factors unique to New Zealand in an attempt to explain the increasing prevalence of asthma around the world. It may be more productive to direct future research towards identifying factors that the major English-speaking countries have in common. PMID- 11482330 TI - Excessive workloads for junior doctors. PMID- 11482331 TI - General practice obstetrics. PMID- 11482332 TI - Quality assurance in colposcopy using video capture and the internet for individual audit. PMID- 11482333 TI - Diabetes is still unrecognised in the 1990's: report of a hospital audit. PMID- 11482334 TI - Myocardial infarction and rupture. PMID- 11482335 TI - Retinal pigment epithelial tear with vitreomacular attachment: a novel pathogenic feature. AB - BACKGROUND: The development of tears of the retinal pigment epithelium (RPE) has classically been described with or without choroidal neovascularization (CNV) or after laser treatment. Tangential shear forces within the RPE or CNV are usually considered to cause the dehiscence. METHODS: Three patients with CNV and spontaneous RPE tear and additional vitreomacular traction were examined by fluorescein angiography (FA), optical coherence tomography (OCT) and kinetic ultrasound. RESULTS: From the pre-tear to the tear stage a sudden decrease in vision was observed. Fluorescein angiographic images demonstrated RPE-tear formation with blocked filling in the area of the contracted RPE and a well demarcated hyperfluorescence in the bed of the torn RPE. OCT-scans demonstrated vitreomacular traction at the foveal area in all three cases. Kinetic ultrasound revealed vitreous attachments at the optic disc and fovea. CONCLUSION: Magnitude, variation of mechanical forces, and the continuous shear stress of the aged vitreous gel transmitted across vitreoretinal attachments may cause a chronic stimulus to retina and RPE. Vitreomacular traction may contribute to the subsequent formation of RPE tears via mechanical or cell mediator pathways. PMID- 11482336 TI - Surgery for nystagmus related head turn: Kestenbaum procedure and artificial divergence. AB - PURPOSE: An abnormal head posture adopted to reduce a nystagmus can be treated by Kestenbaum surgery or by creating an exodeviation which is compensated by convergence (artificial divergence). We evaluated the effects of Kestenbaum surgery and artificial divergence surgery in a retrospective study. METHODS: Seventy-eight patients who received surgery for horizontal head turn (HT) due to nystagmus were included in the study. Exclusion criteria were previous extraocular muscle surgery, strabismus, lack of binocular vision, and cerebral disease. The millimetres of surgery and the findings before and 3 months after operation [HT, binocular visual acuity (VA), binocular vision (BV)] were evaluated. The patients were divided into three groups: the Kestenbaum group, the artificial divergence group, and the artificial divergence plus Kestenbaum group. RESULTS: Of the 78 patients, aged 3-68 years, 52 had HT to the left side and 47 were male. In the Kestenbaum group (n=31), the median (10% and 90% quantile) HT was 30 degrees (range 20-40 degrees). A total of 28 mm (range 20-40 mm) surgery reduced the HT to 10 degrees (0-30 degrees). The efficacy of surgery was 1.4 degrees HT reduction per millimetre surgery on one eye (range 0.4-2.5 degrees). Four patients received further surgery. In the artificial divergence group (n=27), 10 mm (range 7-12 mm) surgery reduced the HT of 30 degrees (range 25-40 degrees) to 5 degrees (range 0-20 degrees). Seven patients received further surgery. In the artificial divergence plus Kestenbaum group (n=20), the HT was 30 degrees (range 25-40 degrees). A total of 29 mm (range 21-37 mm) surgery reduced the HT to 7 degrees (range -5 degrees to 15 degrees). No further surgery was performed. Postoperatively, the maximum VA and BV was available without large HT, but an increase in the absolute VA and BV could not be proved. CONCLUSION: Artificial divergence is preferable or should be combined with Kestenbaum surgery, if possible. Kestenbaum surgery alone has an effect/dose ratio similar to recess-resect surgery for strabismus. Thus, to correct x degrees HT, 2/3x mm surgery on each eye is adequate. PMID- 11482337 TI - Agreement between ophthalmologists and optometrists in optic disc assessment: training implications for glaucoma co-management. AB - BACKGROUND: Many studies have estimated observer variability for optic disc assessment among experts, but there are few data on agreement between optometrists and ophthalmologists. The aim of this study is to report inter- and intraobserver agreement among optometrists and ophthalmologists in optic disc assessment and discuss the implications for glaucoma co-management. METHODS: Twelve observers (six optometrists and six ophthalmologists) graded 48 stereo pairs of optic disc photographs from 48 patients on two separate occasions. Observers graded the vertical C/D ratio, the narrowest rim width and the presence/absence of disc haemorrhage. Agreement was assessed by calculating the standard deviation (SD) of differences and the kappa statistic, within and between observers. Systematic differences in grading between and within individuals were described by calculating mean differences. RESULTS: Intra observer agreement is "substantial" and significantly better than inter-observer agreement for all disc features (P<0.0001). Intra-observer agreement is comparable for optometrists and ophthalmologists with regard to C/D ratio estimates. Although some optometrists show close agreement with ophthalmologists, overall agreement between optometrists and ophthalmologists is significantly worse than agreement among ophthalmologists alone for vertical C/D ratio (P=0.002) and disc haemorrhage (P=0.02). There are fewer inter-observer mean differences that differ significantly from zero among ophthalmologists (7%) than among optometrists (67%; chi2=11.63, P<0.001) or between optometrists and ophthalmologists (69%; chi2=16.7, P<0.001). There is evidence of systematic under reading of C/D ratios by less experienced optometrists. CONCLUSION: Although individual optometrists can show good agreement with ophthalmologists on disc grading, the variation in individual performance indicates that training and accreditation in disc assessment is an essential prerequisite for participation by optometrists in glaucoma co-management. PMID- 11482338 TI - Comparison of frequency doubling perimetry and standard achromatic computerized perimetry in patients with glaucoma. AB - PURPOSE: To evaluate the reproducibility and efficacy of frequency-doubling perimetry (FDP), a new perimetric method for the detection of functional glaucomatous damage. METHODS: Thirty-four glaucomatous eyes of 34 patients were examined by FDP (using the complete N30 threshold test of the Humphrey Frequency Perimeter) and by standard achromatic computerized perimetry (SAC; using the full threshold program 30-2 of the Humphrey Field Analyzer). Twenty-six of these eyes were examined twice by FDP. Reproducibility of repeated FDP was tested by the difference and the average of mean deviations (MD). Efficacy was studied by comparison of FDP and SAC using MD, as well as threshold results of FDP and corresponding SAC-tested areas. Spearman's rank correlation was calculated. RESULTS: A significant correlation of repeated FDP examinations was found regarding MD [r=0.94, P<0.00001 (n=26)]. The reproducibility of FDP was independent of the glaucomatous damage [r=0.04, P=0.85 (n=26)], based on the correlation of the difference, and the average of MD. The MD of FDP and that of SAC showed a significant correlation [r=0.87, P<0.00001 (n=34)]. In advanced glaucoma, FDP tended to yield lower measurement values than SAC. Regarding corresponding areas, there was a significant correlation between threshold results of FDP-tested areas and the corresponding SAC testpoints [r=0.92, P<0.00001 (n=34)]. The conformity decreased from the peripheral to the central tested visual field. CONCLUSIONS: Sufficient reproducibility of FDP could be demonstrated in early and advanced glaucomatous visual field defects. Conformity between FDP and SAC was shown in terms of MD and threshold results of FDP-tested areas and corresponding SAC testpoints. PMID- 11482339 TI - Reliability and reproducibility of corneal endothelial image analysis by in vivo confocal microscopy. AB - BACKGROUND: The aim of our study was to evaluate the reliability and reproducibility of in vivo slit-scanning corneal microscopy by endothelial analysis. METHODS: We examined 12 normal eyes of 12 persons. We analysed intraobserver differences in group I and interobserver differences in group II by manual and automatic evaluation of endothelial images. We also compared the methods of analysis in the two groups. We observed endothelial density, mean cell area, coefficient of variation and number of cells within the frame. RESULTS: In group I we found no significant difference between the observed parameters either by automatic or manual analysis. In group II no significant difference between the two observers was found using automatic analysis. On manual analysis, however, we observed a significant difference between endothelial density and the number of evaluated cells (P=0.0034 and P=0.0028). We observed a significant difference (P<0.001) between automatic and manual analysis concerning each parameter, with a rather tight linear correlation (Pearson correlation between 0.76 and 0.94). CONCLUSION: Automatic endothelial analysis was reliable and well reproducible in both--intraobserver and interobserver--groups. By manual evaluation, the clinical significance of interobserver differences can be disregarded. The differences between automatic and manual methods of analysis can be traced back to measurement technical reasons. We observed tight linear correlation between parameters. The data can be described well by linear regression. In vivo slit-scanning corneal microscopy may be an alternative to specular microscopic analyses for clinical use. PMID- 11482340 TI - Rigid gas-permeable contact lens fitting in LASIK patients for the correction of multifocal corneas. AB - BACKGROUND: Laser in situ keratomileusis (LASIK) has recently become the most commonly performed refractive surgery procedure. Results are promising in correcting low to moderate myopia. Most complications occur during the surgeon's learning curve. One of the complications is a decentration of the ablated area that causes monocular diplopia and a nocturnal halo phenomenon due to a multifocality of the corneal surface overlying the entrance pupil. The corneal shape is significantly altered after LASIK. We evaluate the efficacy of rigid gas permeable contact lens designs and fitting techniques used in eight eyes with multifocal LASIK ablations to correct haloes and impaired night vision complaints. METHODS: We used large-diameter tetra-curved rigid gas-permeable (RGP) contact lenses for visual recovery in eight eyes of seven LASIK patients. We used the power on the transition zone of the corneal topographic map, 0.2 mm outside the ablated refractive area, for selection of the back optic zone radius of the RGP contact lenses. Procedures for lens fitting are described. Visual acuity (high-contrast logarithm of the minimum angle of resolution, LogMAR) was measured before- and 6 months after contact lens fitting. RESULTS: Large-diameter tetra-curve RGP lenses with a mean diameter of 11.85 (SD 0.16) mm were successfully used in LASIK patients with multifocal corneas. Mean best spectacle corrected visual acuity was +0.3 LogMAR (SD 0.19; in Snellen equivalent, 20/40) and improved significantly with the contact lenses to +0.08 LogMAR (SD 0.11; in Snellen equivalent, 20/25, P=<0.01). During the follow-up period of 16.7 months, the average daily wearing time of the lenses was 12.5 h. Contact lenses with a standard geometry were not useful due to excessive movement and inadequate centration. CONCLUSIONS: Contact lenses with large diameters, in combination with selection of the back optic zone radius 0.2 mm peripheral of the refractive ablation zone, facilitate contact lens fitting to restore best-corrected visual acuity in LASIK patients with multifocal corneas. PMID- 11482341 TI - Systemic mycophenolate mofetil in comparison with systemic cyclosporin A in high risk keratoplasty patients: 3 years' results of a randomized prospective clinical trial. AB - BACKGROUND: With the use of systemic cyclosporin A (CsA), graft prognosis after high-risk penetrating keratoplasty has improved considerably. However, the application of CsA is limited owing to a variety of severe side effects. In this prospectively randomized study mycophenolate mofetil (MMF), a safe and efficient immunosuppressive medication after renal transplantation, was compared with CsA after high-risk penetrating keratoplasty. METHODS: Twenty-nine high-risk keratoplasty patients were treated with MMF 2x 1 g daily; another 27 patients received CsA, aiming at blood trough levels of 120-150 ng/ml. Systemic immunosuppression was scheduled for 6 months. In both groups oral corticosteroids (fluocortolone 1 mg/kg) were administered for 3 weeks postoperatively. RESULTS: During the first year after operation, no graft failure was recorded. Two years postoperatively 92%/82% and 3 years postoperatively 74%/69% of grafts were clear in the MMF and CsA group, respectively (Kaplan Meier P=0.33, logrank test). In total, two graft failures were recorded in the MMF group and four in the CsA group. Three years postoperatively 53% of the grafts were rejection-free in the MMF group and 73% in the CsA group (Kaplan Meier P=0.46, log-rank test). Eight endothelial immune reactions were observed in the MMF group (three under systemic immunosuppression/five thereafter; six mild/two severe) and five in the CsA group (three under systemic immunosuppression/two thereafter; three mild/two severe). Side effects occurred in six patients under MMF and 11 under CsA. CONCLUSIONS: Concerning efficacy, no statistically significant difference between systemic MMF and systemic CsA administered for 6 months after high-risk penetrating keratoplasty could be shown. Systemic MMF was proven to be at least as safe as CsA. The main mechanism in improving graft survival is a shift from severe to milder endothelial immune reactions, as already demonstrated for CsA. Thus, MMF may become an alternative to CsA for immunosuppression after penetrating high risk keratoplasty. About 2 years postoperatively, pharmacologically induced relative immunological tolerance slowly decreases. Therefore, long-term administration of systemic MMF should be evaluated in further studies. PMID- 11482342 TI - Combined use of partially fluorinated alkanes, perfluorocarbon liquids and silicone oil: an experimental study. AB - BACKGROUND: Partially fluorinated alkanes (FALKs) are a new class of substances which can be used in vitreoretinal surgery as an intraoperative tool and as a long-term tamponade. The aim of this in vitro study was (1) to investigate the solubility of FALKs in silicone oil during direct exchange, (2) to study their combined use and solubility in PFCLs, (3) to evaluate their lipophilic properties and (4) to investigate the possibility of preparing "heavy silicone oil". METHODS: (1) Four different FALKs (F6H6, F6H8, O44 and O62) were directly exchanged with silicone oil (5,000 mPas). The dissolved amount of fluorocarbons in the removed silicone oil was determined by gas chromatography and by gravimetry. Furthermore, the diffusion phenomena during the exchange process were studied. (2) The behaviour of FALKs in PFCLs was investigated and the solubility of the resulting mixtures in silicone oil was measured. (3) The solubility of FALKs and PFCLs in native olive oil was analysed. (4) Different FALKs were added to silicone oil and measurements of the resulting specific gravity and the viscosity were performed. RESULTS: (1) FALKs dissolved in silicone oil up to the following values: F6H6=45 m%, F6H8=54 m%, 044=100 m%, O62=18 m%. (2) FALKs dissolved in PFCL, thereby changing the physicochemical properties of PFCL depending on the type of FALK and ratio used. (3) The lipophilic properties of FALKs and PFCLs could be characterized by their dissolution in native olive oil (F6H8=23.4 m%, 044=16.7 m%, F6H6=12.3 m%, 062=5.3 m%, PFD=1.1 m%, PFO=0.6 m%). (4) It was possible to prepare "heavy silicone oil" e.g. by adding 30 vol% F6H8, resulting in a specific gravity of 1.08 g/ml, or by adding 80 vol% 044, resulting in a specific gravity of 1.25 g/ml, but decreasing the viscosity of the mixtures dramatically. CONCLUSION: (1) If FALKs are used as an intraoperative tool, a direct exchange with silicone oil should be avoided owing to their capacity to dissolve in silicone oil, resulting in a mixture with unpredictable properties. (2) A combined use with PFCLs and silicone oil is possible, if the right ratio is chosen. (3) The solubility of FALKs in native olive oil may be an indicator for their tissue penetration and may render feasible their use as a long-term tamponade. (3) "Heavy silicone oil" preparation using FALKs is possible, but the mixture needs further evaluation in terms of emulsification, mobilization of oligosiloxanes, tissue penetration and long-term stability. PMID- 11482343 TI - Topographic analysis of astigmatism induced by scleral shortening in pig eyes. AB - BACKGROUND: Corneal astigmatism is a severe postoperative problem in foveal translocation surgery. We evaluated the corneal astigmatism induced by scleral shortening in pig eyes in vitro. METHODS: We created three sizes of scleral shortening in pig eyes and examined the preoperative and postoperative corneal astigmatism. The three sizes of scleral shortening were; 6 mm x 12 mm, 9 mm x 12 mm, and 6 mm x 16 mm (radial x circumferential). The shortenings were created 11 mm from the limbus with 10 eyes in each group. Videokeratographic measurements were performed using the CAS System 2000, preoperatively and postoperatively, and the astigmatism caused by the scleral shortening was evaluated. RESULTS: The surgically-induced astigmatism was 2.1 +/- 1.2 diopters (D) in the 6 mm x 12 mm group, 5.2+/-1.5 D in the 9 mm x 12 mm group, and 3.7+/-1.0 D in the 6 mm x 16 mm group. Corneal astigmatism caused by scleral shortening depended on both the radial and circumferential shortening. Pre- and postoperative topographic corneal maps showed an irregular astigmatism pattern (lazy bowtie pattern). Because the central zone of the cornea showed a relatively regular astigmatism, the corneal astigmatism induced by scleral shortening did not affect the predicted corneal acuity. CONCLUSIONS: In foveal translocation surgery with scleral shortening, an excessive scleral resection in the radial direction can cause clinically intolerable regular and irregular astigmatism. Minimal scleral shortening that will satisfy the required translocated distance is recommended to reduce the risk/benefit ratio. PMID- 11482344 TI - Persistent indocyanine green (ICG) fluorescence 6 weeks after intraocular ICG administration for macular hole surgery. AB - BACKGROUND: Macular hole surgery including vitrectomy and peeling of epiretinal membranes and the internal limiting membrane (ILM) has become a standard procedure in retinal surgery. Poor visualization of epiretinal membranes and the ILM is an obstacle to successful surgery. Recently, indocyanine green (ICG) has been reported to be a helpful intraocular substance in identifying these membranes. METHODS: In a case of stage IV macular hole, epiretinal membranes and ILM were intraoperatively stained with three drops of 1:9 diluted ICG. After 1 min incubation the vitreous cavity was rinsed with Ringer's lactate solution, and the membranes were peeled. Autologous thrombocytes were applied to the macular hole, and the eye was endotamponaded with 20% SF6 gas. Six weeks postoperatively, visual acuity was measured and fundus photographs and autofluorescence images, as well as a multifocal ERG, were obtained. RESULTS: Intraoperatively, the ILM could be nicely visualized by ICG, which allowed immediate peeling. Six weeks after surgery, the visual acuity had improved from 0.1 to 0.7 and the macular hole was closed. Autofluorescence imaging at 795 nm revealed a strong signal. Multifocal ERG recording showed regular amplitudes. CONCLUSION: ICG as an intraocular tool for staining of the ILM is helpful in macular hole surgery. We did not observe any negative effect on retinal function; however, we were surprised to identify traces of ICG in retinal fluorescein angiography images 6 weeks postoperatively. PMID- 11482345 TI - Typical ocular findings in a patient with multiple endocrine neoplasia type 2b syndrome. AB - BACKGROUND: Multiple endocrine neoplasia (MEN) type 2b syndrome is accompanied by typical ocular findings; however, the disease is often only diagnosed at an advanced stage by symptoms of C-cell carcinoma or pheochromocytoma and is then fatal in most cases. Therefore, the importance of ophthalmic assessment in making the diagnosis has to be stressed. METHODS: The history and ocular findings of a patient with MEN 2b syndrome are described, and a brief overview of the syndrome is given. RESULTS: Slit-lamp examination showed extremely thickened corneal nerves as well as multiple small plexiform and nodular subconjunctival tumors. Both eyes also displayed thickened upper and lower eyelids. A molecular genetic study of the RET proto-oncogene showed a heterozygous ATG to ACG mutation in codon 918 of exon 16. CONCLUSION: Greatly thickened corneal nerves and subconjunctival tumors may be the first hint of MEN 2b. Whenever greatly thickened corneal nerves are detected, MEN 2b must be ruled out. PMID- 11482346 TI - Nonviral gene therapy and its delivery systems. AB - Nonviral gene therapy has significant clinical potential, yet its therapeutic utility has been hindered by low transfection efficiency due to a combination of extracellular and intracellular barriers. Recent developments in formulation and delivery methodology have allowed a number of advances toward high efficiency gene delivery to various cell types and organs. In particular, the extracellular and intracellular pharmacokinetics of plasmid DNA trafficking are better understood in a number of cell systems. Using cationic lipid or polymers (often with receptor targeting), more than 10(5) plasmids can be delivered to a single cell. Endosomolytic agents promote endosome disruption, and include: weak bases, proton-sponge polymers, fusogenic peptides, viral particles, and photosensitizing compounds. Both classical and nonclassical nuclear localization signal (NLS) peptides have also been tested for enhancement of the probability of nuclear import events, a major rate-limiting step in DNA delivery to nondividing cells. For example, the M9 sequence from heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) protein, a nonclassical NLS, has been found to increase gene expression level by more than 10 to 150-fold in a variety of cell types. This review will concentrate on the current understandings of the basic mechanisms of nonviral gene delivery and new approaches in the field. PMID- 11482347 TI - Antibody engineering for targeted therapy of cancer: recombinant Fv-immunotoxins. AB - Recombinant Fv-immunotoxins are a new class of biologic anticancer agents composed of a recombinant antibody fragment linked to a very potent bacterial toxin. These potent molecules are designed to specifically bind and kill cancer cells that express a specific target antigen on their cell surface. Recombinant Fv-immunotoxins are an excellent example for the concept of rational drug design. They combine the progress in understanding cancer biology, -the recent knowledge on the mechanisms of malignant transformation and the special properties of cancer cells, -with the enormous developments in recombinant DNA technology and antibody engineering. Recombinant Fv immunotoxins were developed for solid tumors and hematological malignancies and have been characterized intensively for their biological activity in vitro and in vivo in animal models. The excellent in vitro and in vivo activities of recombinant Fv-immunotoxins have lead to their pre clinical development and to the initiation of clinical trial protocols. Recent trials have demonstrated potent clinical efficacy in patients with malignant diseases that are refractory to traditional modalities of cancer treatment. It is thus suggested that this strategy can be developed into a separate modality of cancer treatment with the basic rationale of specifically targeting cancer cells on the basis of their unique surface markers combined with potent effective biological toxic agents that directly kill the cancer cell. Efforts are now being made to improve the current molecules and to develop new agents with better clinical efficacy. In this review, we will describe the rationale in designing Fv immunotoxins and will review current progress made in using these agents for cancer treatment. PMID- 11482348 TI - Genetically modified viruses: vaccines by design. AB - Vaccination has been one of the most successful and cost-effective health interventions ever employed. One disease (smallpox) has been eradicated, another (poliomyelitis) should disappear early in the new millennium and a third (measles) should follow shortly after. Conventional vaccines usually depend on one of three development processes, attenuation of virulent organisms (by passage in cell culture and/or experimental animals), killing of virulent organisms (by chemical inactivation) or the purification of immunogenic molecules (either proteins or carbohydrates) from whole organisms. These traditional processes, although serendipitous and poorly understood, have produced effective pharmaceutical products which give excellent protection against diseases such as smallpox, rabies, measles, yellow fever, tetanus and diphtheria. In spite of these successes however, the application of these protocols have failed to produce safe and efficacious vaccines against other infectious diseases which kill or maim tens of millions of people every year. The most important of these are malaria, AIDS, herpes, dengue fever and some forms of viral hepatitis. Consequently, fundamentally new technologies are required to tackle these important infections. One of the most promising has been the development of genetically modified viruses. This process normally involves taking a proven safe and efficacious vaccine virus, such as vaccinia or adenovirus, and modifying its genome to include genes coding for immunogenic proteins from other viruses such as HIV or measles. This review will describe the generation of such novel vaccine vectors and compare their advantages and shortcomings. In addition the literature describing their use as experimental vaccines will also be reviewed. PMID- 11482349 TI - Small heat shock proteins (sHSPs) as potential drug targets. AB - Small heat shock proteins (sHSPs) belong to a family of 12- to 43-kDa proteins that are ubiquitous and are largely conserved in amino acid sequence among all organisms. The principal heat-shock proteins that have chaperone activity (that is, they protect newly made proteins from misfolding) belong to five conserved classes: HSP100, HSP90, HSP70, HSP60 and the small heat-shock proteins (sHSPs). The sHSPs (which include alpha crystallin) can form large multimeric structures and have a wide range of cellular functions, including endowing cells with thermotolerance in vivo and being able to act as molecular chaperones in vitro; sHSPs do this by forming stable complexes with folding -or unfolding- intermediates of their protein substrates, probably the molten globule. This paper includes: a brief survey of the chaperone family, the small heat shock protein superfamily, transcription of sHSPs, sequence comparisons and structural models of small heat shock proteins--structural elements as potential drug targets, sHSPs as chaperone-like proteins, alpha crystallin chaperone-like activity, conformational diseases--the role of alpha crystallin small heat shock protein superfamily proteins, post-translational modification and useful pharmacological agents. Functionality of small heat shock proteins--targets and diseases where pharmacologically active agents are of importance, alpha crystallin--small heat shock proteins and prion diseases: specific targets for diagnostic tests and drug development, details of some specific small heat shock proteins as drug targets, structural and functional implications for treatment. PMID- 11482350 TI - The responsiveness of the Action Research Arm test and the Fugl-Meyer Assessment scale in chronic stroke patients. AB - The responsiveness of the Action Research Arm (ARA) test and the upper extremity motor section of the Fugl-Meyer Assessment (FMA) scale were compared in a cohort of 22 chronic stroke patients undergoing intensive forced use treatment aimed at improvement of upper extremity function. The cohort consisted of 13 men and 9 women, median age 58.5 years, median time since stroke 3.6 years. Responsiveness was defined as the sensitivity of an instrument to real change. Two baseline measurements were performed with a 2-week interval before the intervention, and a follow-up measurement after 2 weeks of intensive forced use treatment. The limits of agreement, according to the Bland-Altman method, were computed as a measure of the test-retest reliability. Two different measures of responsiveness were compared: (i) the number of patients who improved more than the upper limit of agreement during the intervention; (ii) the responsiveness ratio. The limits of agreement, designating the interval comprising 95% of the differences between two measurements in a stable individual, were -5.7 to 6.2 and -5.0 to 6.6 for the ARA test and the FMA scale, respectively. The possible sum scores range from 0 to 57 (ARA) and from 0 to 66 (FMA). The number of patients who improved more than the upper limit were 12 (54.5%) and 2 (9.1%); and the responsiveness ratios were 2.03 and 0.41 for the ARA test and the FMA scale, respectively. These results strongly suggest that the ARA test is more responsive to improvement in upper extremity function than the FMA scale in chronic stroke patients undergoing forced use treatment. PMID- 11482351 TI - Gait improvement in unilateral transfemoral amputees by a combined psychological and physiotherapeutic treatment. AB - A conscious therapeutic approach was used combining methods in physiotherapy with psychological awareness to re-educate nine transfemoral amputees during 10 months in outdoor environments. All were rehabilitated trauma or tumour cases, mean age 33 years, and had worn their prostheses for more than 18 months. The method aimed at integrating the prosthesis in normal movements and increasing body awareness. Gait was measured with a three-dimensional motion analysis system. Self-selected comfortable and brisk gait speed increased from mean 0.95 m/s and 1.29 m/s before to 1.40 m/s and 1.65 m/s after treatment, respectively. The results remained at a 6-month follow-up. Before treatment three participants used walking-aids and all had problems with low-back pain. After treatment none needed walking-aids and almost all low-back pain had disappeared. Seven participants learnt to jog. Results indicate that this new approach may add skills, mostly on participation level, to lead a relatively normal life. PMID- 11482352 TI - Distress in everyday life in people with poliomyelitis sequelae. AB - The prevalence of distress in aspects of perceived health and its relation to involvement of poliomyelitis sequelae were studied with the Nottingham Health Profile (NHP) in 113 outpatients (mean age 57 years). The leisure and employment situation was also recorded. Most distress was found in the NHP dimensions physical mobility, pain and energy, and least distress in social isolation. Most health-related problems were reported in housework, employment and leisure. Three quarters of the persons were satisfied with their leisure, although many of them had problems. Fifty-nine per cent of the subjects of working age were in gainful employment, and no difference in employment rate due to the distribution of polio involvement was found. In comparison with norm values for the respective age groups, the subjects with poliomyelitis sequelae aged below 45 and 45-65 years had more distress in a larger number of NHP dimensions than older subjects. PMID- 11482353 TI - Assessment of chronic pain behaviour: reliability of the method and its relationship with perceived disability, physical impairment and function. AB - The aim of the present study was to develop a reliable assessment of pain behaviour performed during the execution of a range of functional assessment measures. For the initial reliability study 18 subjects (consecutive referrals) were assessed. Subjects were observed and videotaped during a variety of physical tasks and demonstrations of pain behaviour were recorded; the videotapes were scored by two independent observers on two occasions. The relationships between pain behaviour, distress and physical function and impairment were also investigated in a group of 51 patients with chronic back pain. Self-report of disability and pain intensity were assessed using the Finnish version of Oswestry disability questionnaire and the pain visual analogue scale (VAS). Depression and somatic perception were assessed using the modified Zung and modified somatic perception questionnaire. The Tampa scale for kinesiophobia was used to evaluate fear of movement and (re)injury. The results of the intra- and interobserver reliability study demonstrate good to excellent levels of agreement. The exception was facial expression (kappa 0.29), which was excluded from the final instrument. There was a strong correlation between pain behaviour and subjective pain report and disability (p < 0.01). The correlations between total pain behaviour and performance of physical function tasks is striking (p < 0.01). Subjective disability was analysed by means of multiple regression analysis. Pain measured on the VAS was the most important variable explaining 36% of the variance, pain behaviour and pain combined explained 48% of the variance for self reported disability. In conclusion, this functional videobased assessment of pain behaviour is a reliable measure of pain behaviour. The total scores for pain behaviour correlate with tasks that involve the back; tests involving upper limbs were not affected. This test is suitable for the assessment of those with pain problems specifically involving the back. Furthermore, in the group studied pain and pain behaviour were the two most important determinants of self-reported disability. PMID- 11482354 TI - The influence of the viewing angle on neck-load during work with video display units. AB - Ergonomic measures have been found to reduce load-related trouble from the neck and-shoulders during visual display unit (VDU) work. An important question is the height at which the screen should be placed to give the lowest possible load. Should it be placed at eye-level or below? The aim of the present study was to investigate whether there was any difference in external loading moments of force about the C7-T1 segment when the VDU-operators had a viewing angle of 20 degrees below the horizontal plane as compared to 3 degrees above the horizontal. Eight secretaries were videofilmed in the sagittal plane in the two work postures during simulated work. The loading moment was calculated from the film. It was significantly lower at viewing angle 3 degrees above the horizontal than at 20 degrees below the horizontal, both at the beginning (1.3 vs 2.2 nm) and at the end (1.4 vs 2.1 nm) of the film sequences (p < 0.05). PMID- 11482355 TI - Measures of order consistency between paired ordinal data: application to the Functional Independence Measure and Sunnaas index of ADL. AB - The aim was to evaluate the inter-item consistency between corresponding items in the Functional Independent Measure (FIM) and Sunnaas index of ADL (SI) by means of a statistical approach that takes account of the nonmetric properties of ordinal data. The ADL dependence in 204 stroke patients was assessed by interview 3 months after the onset of stroke. The result showed that the inter-item consistency between the FIM and SI were high for many items, but operational differences between some FIM and SI items were also identified. The statistical evaluation demonstrated that some of the ordered categorical levels of the seven point item scales in the FIM could be aggregated into four levels without loss of information. PMID- 11482356 TI - Early rehabilitation after severe brain injury: a French perspective. AB - Severely brain-injured patients often suffer from disabilities and psycho-social handicaps. Early rehabilitation aims at improving their motor and functional recovery while preventing or treating complications as soon as possible. In this review we look at some issues encountered in early rehabilitation. We illustrate our discussion with data from 876 French traumatic brain injury patients admitted over the course of 1 year at 18 rehabilitation units that were asked for details of their current practice. Preservation of vital functions follows standardized protocols, but rehabilitation is more controversial. Few controlled trials are available. Good agreement exists among clinicians about prevention of orthopedic complications and treatment for spasticity. However, little consensus exists concerning treatment of non-pyramidal hypertonia and spasms or about procedures that can be undertaken to improve arousal from a coma or vegetative state. Finally, we look at other specific issues of early rehabilitation, namely prediction of outcome, psychological difficulties of patients and their families, efficiency and cost-effectiveness. PMID- 11482357 TI - A new hybrid spring brake orthosis for controlling hip and knee flexion in the swing phase. AB - In this study it is proposed that active contraction of muscles might be artificially replaced by a spring brake orthosis (SBO) to provide near-natural knee and hip swing phase trajectories for gait in spinal cord injured subjects. The SBO is a new gait restoration system in which stored spring elastic energy and potential energy of limb segments are utilized to aid gait. It is also shown that hip flexion can be produced without the need for withdrawal reflex, hip flexor stimulus or any mechanical actuator at the hip. A hip flexion angle of 21 degrees was achieved by a nonimpaired subject wearing a prototype orthosis. PMID- 11482358 TI - Modulation and vectorial summation of the spinalized frog's hindlimb end-point force produced by intraspinal electrical stimulation of the cord. AB - The ability to produce various force patterns at the ankle by microstimulation of the gray matter of the spinal cord was investigated in spinalized frogs. We evaluated the recruitment properties of individual spinal sites and found that forces increase linearly with activation level in the low-force range studied, while the structure of the force pattern remains invariant. We also measured the responses produced by coactivation of two spinal sites activated at two pairs of stimulation levels. Responses were measured at the mechanical level by recording forces at the ankle; and, at the muscular level by recording the electromyographic (EMG) activity of 11 hindlimb muscles. We found that for both pairs of activation, the forces under coactivation were the scaled vectorial summation of the individual responses. At the muscular level, rectified and integrated EMGs also summated during coactivation. Numerous force patterns could, thus, be created by the activation of a few individual sites. These results suggest that microstimulation of the circuitry of the spinal cord (higher order neurons than the motoneurons) holds promise as a new functional neuromuscular stimulation (FNS) technique for the restoration of multi-joint movements. PMID- 11482359 TI - Electrodeposited iridium oxide for neural stimulation and recording electrodes. AB - Iridium oxide films formed by electrodeposition onto noniridium metal substrates are compared with activated iridium oxide films (AIROFs) as a low impedance, high charge capacity coating for neural stimulation and recording electrodes. The electrodeposited iridium oxide films (EIROFs) were deposited on Au, Pt, PtIr, and 316 LVM stainless steel substrates from a solution of IrCl4, oxalic acid, and K2CO3. A deposition protocol involving 50 potential sweeps at 50 mV/s between limits of 0.0 V and 0.55 V (versus Ag AgCl) followed by potential pulsing between the same limits produced adherent films with a charge storage capacity of >25 mC/cm2. Characterization by cyclic voltammetry and impedance spectroscopy revealed no differences in the electrochemical behavior of EIROF on non-Ir substrates and AIROF. The mechanical stability of the oxides was evaluated by ultrasonication in distilled water followed by dehydration and rehydration. Stability under charge injection was evaluated using 200 micros, 5.9 A/cm2 (1.2 mC/cm2) cathodal pulses. Loss of iridium oxide charge capacity was comparable for AIROFs and the EIROFs, ranging from 1% to 8% of the capacity immediately after activation or deposition. The EIROFs were deposited and evaluated on silicon microprobe electrodes and on metallized polyimide electrodes being developed for neural recording and stimulation applications. PMID- 11482360 TI - Spectral characteristics of visually induced postural sway in healthy elderly and healthy young subjects. AB - Spectral analysis and time-frequency analysis were applied to anterior-posterior (A-P) center-of-pressure (COP) data collected during quiet stance and sinusoidal (0.25 Hz) moving visual scene perturbations from sixteen healthy elderly subjects and thirteen healthy young subjects. While the total energy of COP was larger in the elderly subjects than in the young subjects, energy-normalized spectra of the entire COP time series were similar between the two populations. Further time dependent analysis of the spectral characteristics showed that the time-varying spectra were also similar, as revealed by time-frequency analysis, although some differences were observed. Specifically, time-dependent mean frequency and bandwidth responses to the stimulus showed similar increases in both young and elderly subjects with the introduction of the stimulus. Furthermore, time frequency analysis showed that both groups exhibited an initial increase in sway at the stimulus frequency that then declined as the sinusoidal scene movement progressed. This reduction in sway during visual stimulation may indicate an "adaptation" to constant frequency visual perturbation that is invariant with age. This finding is in contrast to results reported recently for patients with vestibular impairments, who appear to lack such adaptation. PMID- 11482361 TI - Quantification of the UPDRS Rigidity Scale. AB - In the clinical setting, parkinsonian rigidity is assessed using subjective rating scales such as that of the Unified Parkinson's Disease Rating System (UPDRS). However, such scales are susceptible to problems of sensitivity and reliability. Here, we evaluate the reliability and validity of a device designed to quantify parkinsonian rigidity at the elbow and the wrist. The method essentially quantifies the clinical examination and employs small sensors to monitor forces and angular displacements imposed by the clinician onto the limb segment distal to the joint being evaluated. Force and displacement data are used to calculate elastic and viscous stiffnesses and their vectorial sum, mechanical impedance. Interexaminer agreement of measures of mechanical impedance in subjects with Parkinson's disease was comparable to that of clinical UPDRS scores. Examiners tended to overrate rigidity on the UPDRS scale during reinforcement manoeuvres. Mechanical impedance was nonlinearly related to UPDRS ratings of rigidity at the elbow and wrist; characterization of such relationships allows interpretation of impedance measurements in terms of the clinical rating scales. PMID- 11482362 TI - A neural network model for optimizing vowel recognition by cochlear implant listeners. AB - Due to the variability in performance among cochlear implant (CI) patients, it is becoming increasingly important to find ways to optimally fit patients with speech processing strategies. This paper proposes an approach based on neural networks, which can be used to automatically optimize the performance of CI patients. The neural network model is implemented in two stages. In the first stage, a neural network is trained to mimic the CI patient's performance on the vowel identification task. The trained neural network is then used in the second stage to adjust a free parameter to improve vowel recognition performance for each individual patient. The parameter examined in this study was a weighting function applied to the compressed channel amplitudes extracted from a 6-channel continuous interleaved sampling (CIS) strategy. Two types of weighting functions were examined, one which assumed channel interaction, and one which assumed no interaction between channels. Results showed that the neural network models closely matched the performance of five Med-EI/CIS-Link implant patients. The resulting weighting functions obtained after neural network training improved vowel performance, with the larger improvement (4%) attained by the weighting function which modeled channel interaction. PMID- 11482363 TI - Rapid prototyping of an EEG-based brain-computer interface (BCI). AB - The electroencephalogram (EEG) is modified by motor imagery and can be used by patients with severe motor impairments (e.g., late stage of amyotrophic lateral sclerosis) to communicate with their environment. Such a direct connection between the brain and the computer is known as an EEG-based brain-computer interface (BCI). This paper describes a new type of BCI system that uses rapid prototyping to enable a fast transition of various types of parameter estimation and classification algorithms to real-time implementation and testing. Rapid prototyping is possible by using Matlab, Simulink, and the Real-Time Workshop. It is shown how to automate real-time experiments and perform the interplay between on-line experiments and offline analysis. The system is able to process multiple EEG channels on-line and operates under Windows 95 in real-time on a standard PC without an additional digital signal processor (DSP) board. The BCI can be controlled over the Internet, LAN or modem. This BCI was tested on 3 subjects whose task it was to imagine either left or right hand movement. A classification accuracy between 70% and 95% could be achieved with two EEG channels after some sessions with feedback using an adaptive autoregressive (AAR) model and linear discriminant analysis (LDA). PMID- 11482364 TI - Real-time gait event detection for paraplegic FES walking. AB - A real-time method for the detection of gait events that occur during the electrically stimulated locomotion of paraplegic subjects is described. It consists of a two-level algorithm for the processing of sensor signals and the determination of gait event times. Sensor signals and information about the progression of the stimulator though its pre-specified stimulation "pattern" are processed by a machine intelligence (fuzzy logic) algorithm to determine an initial estimate of the patient's current phase of gait. This is then reviewed and modified by a second algorithm that removes spurious gait estimates, and determines gait event times. These gait event times are known to the system within approximately one-half of a gait cycle. The resulting gait event detection system was successfully evaluated on three subjects. Detection accuracy is not adversely affected by day-to-day gait variability. This work resolved technical and practical issues that previously limited the real time application of these methods. In particular, cosmetically acceptable insole force transducers were used. This gait event detector is designed for use in a real time controller for the automatic adjustment of the intensity and timing of stimulation while the subject is walking using functional electrical stimulation (FES). PMID- 11482365 TI - Biomimetic finger control by filtering of distributed forelimb pressures. AB - A linear filter was developed for decoding finger commands from volitional pressures distributed within the residual forelimb. Filter parameters were based on dynamic pressures recorded from the residual limb within its socket, during specific finger commands. A matrix of signal features was derived from eight dimensional (8-D) pressure vectors, and its pseudoinverse comprised the filter parameters. Results with amputees showed that the filter could discriminate specific finger flexion commands, suggesting that pressure vector decoding (PVD) can provide them with biomimetic finger control. PMID- 11482366 TI - Analysis of postural perturbation responses. AB - People with cerebellar ataxia lack lower limb coordination and dissipate sway motion slowly and inefficiently after a posture perturbation. We report a practical and low-cost "human resonance frequency test" for both laboratory and clinical use to quantify progress in balance and cerebellar rehabilitation. We assumed that the center-of-pressure (COP) oscillation rate of decay following a standing posture perturbation is directly related to resonance frequency; a more rapidly dissipating COP oscillation about the position of equilibrium indicates, by definition, more efficient postural control. We hypothesized that following successful physical rehabilitation, people with cerebellar degeneration will have a faster rate of decay of the COP response to an external perturbation. Because the COP is modulated by a synergy of trunk and lower limb motion strategies, COP decay rate may be a useful measure of lower limb coordination in people with cerebellar ataxia. The method was applied to three subjects with cerebellar ataxia before and after rehabilitation; there was good agreement between the calculated COP decay rate and conventionally used gait stability parameters providing pilot data for this simple approach. PMID- 11482367 TI - Automated Braille production from word-processed documents. AB - This paper describes a novel method for automatically generating Braille documents from word-processed (Microsoft Word) documents. In particular it details how, by using the Word Object Model, the translation system can map the layout information (format) in the print document into an appropriate Braille equivalent. PMID- 11482368 TI - Implantation of silicon chip microphotodiode arrays into the cat subretinal space. AB - There are currently no therapies to restore vision to patients blinded by photoreceptor degeneration. This project concerns an experimental approach toward a semiconductor-based subretinal prosthetic designed to electrically stimulate the retina. The present study describes surgical techniques for implanting a silicon microphotodiode array in the cat subretinal space and subsequent studies of implant biocompatibility and durability. Using a single-port vitreoretinal approach, implants were placed into the subretinal space of the right eye of normal cats. Implanted retinas were evaluated post-operatively over a 10 to 27 month period using indirect ophthalmoscopy, fundus photography, electroretinography, and histology. Infrared stimulation was used to isolate the electrical response of the implant from that of the normal retina. Although implants continued to generate electrical current in response to light, the amplitude of the implant response decreased gradually due to dissolution of the implant's gold electrode. Electroretinograms recorded from implanted eyes had normal waveforms but were typically 10-15% smaller in amplitude than those in unimplanted left eyes. The nonpermeable silicon disks blocked choroidal nourishment to the retina, producing degeneration of the photoreceptors. The laminar structure of the inner retinal layers was preserved. Retinal areas located away from the implantation site appeared normal in all respects. These results demonstrate that silicon-chip microphotodiode-based implants can be successfully placed into the subretinal space. Gold electrode-based subretinal implants, however, appear to be unsuitable for long-term use due to electrode dissolution and subsequent decreased electrical activity. PMID- 11482369 TI - Walker-assisted gait in rehabilitation: a study of biomechanics and instrumentation. AB - While walkers are commonly prescribed to improve patient stability and ambulatory ability, quantitative study of the biomechanical and functional requirements for effective walker use is limited. To date no one has addressed the changes in upper extremity kinetics that occur with the use of a standard walker, which was the objective of this study. A strain gauge-based walker instrumentation system was developed for the six degree-of-freedom measurement of resultant subject hand loads. The walker dynamometer was integrated with an upper extremity biomechanical model. Preliminary system data were collected for seven healthy, right-handed young adults following informed consent. Bilateral upper extremity kinematic data were acquired with a six camera Vicon motion analysis system using a Micro-VAX workstation. Internal joint moments at the wrist, elbow, and shoulder were determined in the three clinical planes using the inverse dynamics method. The walker dynamometer system allowed characterization of upper extremity loading demands. Significantly differing upper extremity loading patterns were identified for three walker usage methods. Complete description of upper extremity kinetics and kinematics during walker-assisted gait may provide insight into walker design parameters and rehabilitative strategies. PMID- 11482370 TI - The history of Z-VAD-FMK, a tool for understanding the significance of caspase inhibition. AB - Dr. Robert Smith is one of the pioneers in histochemistry. One of his most important achievements is the recognition of proteolysis as a major physiological and pathophysiological process. As a consequence, he developed selective fluorogenic and chromogenic substrates and specific inhibitors of proteases that allow the (histochemical) analysis of protease activity. One of the latest successes is the design of Z-VAD-fluoromethylketone (FMK), the specific caspase inhibitor, that is a key compound for studies on apoptosis. Its development was originally meant for therapeutic use but unforeseen cytotoxicity of a metabolic derivative of the FMK compound disabled its potential as a drug. However, as a tool for fundamental research it is a great success. The history of Z-VAD-FMK is an example of the creative brain and the tireless perseverance of Robert Smith for which histochemistry and cytochemistry owes him so much. This history of Z VAD-FMK is a well-deserved tribute at the occasion of his 70th birthday. PMID- 11482371 TI - Safety in the histochemistry laboratory. AB - Histochemistry and histopathology laboratories are extremely dangerous workplaces owing to the abundance of chemical hazards that constitute major safety and health concerns. A brief review is presented of some of the hazards to which laboratory personnel are exposed. These include fixatives, solvents, embedding media, dyes and stains, and histochemical substrates. Laboratory staff should be fully informed of the dangers and potential long-term health hazards and take the necessary precautionary measures. Despite the abundance of chemical hazards in the histochemistry laboratory, the overall number of work accidents and injuries remains low. PMID- 11482372 TI - Biological labeling and correlative microscopy. PMID- 11482373 TI - Association of soluble guanylate cyclase with the sarcolemma of mammalian skeletal muscle fibers. AB - Previous investigations have shown that NO-producing nitric oxide synthase (NOS) 1 and CO-generating heme oxygenase (HO-2) are associated with the sarcolemma of skeletal muscle fibers in many mammalian species. Despite numerous roles ascribed to NO and possibly also CO in skeletal muscle, a specific receptor for both gases has hitherto not been found in myofibers. Therefore, in the present work the appearance of the alpha1, beta1 and beta2 subunits of soluble guanylate cyclase (sGC), the most commonly known receptor for NO and potentially also CO, was analysed in mammalian skeletal muscles using immunoblotting and immunohistochemistry. Immunoblotting with an antibody against the beta1 subunit of sGC revealed a band of 70 kDa corresponding to the molecular weight of this protein. Immunohistochemistry with antibodies against the alpha1, beta1 and beta2 sGC subunits showed that the larger part of positivity was present in the sarcolemma region of skeletal muscle fibers and colocalized with NOS-1 mainly in type II myofibers and with HO-2 in type I and type II myofibers. For the first time, sarcolemmal association of sGC and its colocalization with NOS-1 generating the sGC-activator NO and with HO-2 producing the potential sGC upregulator CO have been demonstrated in the present study. These results enable a better understanding of the role of NO and CO in myofibers and suggest a so far unknown molecular mechanism for the interaction of sGC with the sarcolemma. PMID- 11482374 TI - Anti-Thy-1 monoclonal antibody with specific reactivity with vascular endothelial cells in rat glomeruli. AB - Inoculation with anti-Thy-1 antibodies (Abs) in rats induces glomerulonephritis resembling human mesangiolytic and/or mesangioproliferative diseases. Some anti Thy-1 monoclonal Abs (mAbs) react with both mesangial and glomerular endothelial cells, whereas others react solely with mesangial cells in rat kidney. These findings suggest that the rat Thy-1 molecule possesses at least 2 variant forms, including a mesangial and a vascular endothelial isoform. However, anti-Thy-1 mAbs with specific reactivity with glomerular endothelial cells have not been available. We describe here a unique anti-rat Thy-1 mAb, TM78-8. The epitope for TM78-8 is closely related, but not identical, to that for OX-7, a commercially available anti-rat Thy-1 mAb. Immunoblotting, immunohistochemistry and immunoelectron microscopy confirm that TM78-8 reacts exclusively with Thy-1 antigens on the surface of vascular endothelial cells in rat glomeruli. TM78-8 may be a suitable marker for rat glomerular endothelial cells as well as for the vascular endothelial isoform of the rat Thy-1 molecule. Intravenous injection of TM78-8 did not induce glomerulonephritis in rats, whereas OX-7 did, indicating that TM78-8 is not nephritogenic. This finding also corresponds with the current consensus that Thy-1 antigens expressed on mesangial cells play an essential role in the development of Thy-1 nephritis. PMID- 11482375 TI - Glucose-6-phosphate dehydrogenase in small intestine of rabbit: biochemical properties and subcellular localization. AB - Biochemical properties and cellular and subcellular distribution patterns of glucose-6-phosphate dehydrogenase (G6PD) were investigated in small intestine of rabbits. The specific activity of G6PD in fresh homogenates of small intestine was 19 +/- 9 IU/g protein. This value did not change significantly after dialysis. The kinetic and electrophoretic properties of the partially purified enzyme were similar to those found in other rabbit tissues. Enzyme histochemical analysis of G6PD activity using the tetrazolium salt method showed high activity in epithelial cells of villi and crypts of Lieberkuhn. The activity in acinar cells of Brunner's glands was lower than that in epithelium, whereas cells of the muscularis externa showed a very low activity. Immunohistochemical analysis showed that the amounts of G6PD protein were lower in the epithelium than in Brunner's glands and muscularis externa. The differences between distribution patterns of activity and protein of G6PD may reflect the presence of inactive enzyme molecules in Brunner's glands and muscularis externa or posttranslational activation of G6PD in epithelium. Electron microscopic immunocytochemical analysis performed with gold-labelled antibodies showed the presence of G6PD protein throughout the cytoplasm and at smooth endoplasmic reticulum in enterocytes. In Paneth cells and cells of Brunner's glands, G6PD was found in the cytoplasm, at rough endoplasmic reticulum and Golgi complex. Immunolabelling was not found in mitochondria or nuclei. Our findings show that G6PD is heterogeneously distributed in cells of the small intestine and that the enzyme is associated with rough and smooth endoplasmic reticulum to support synthetic functions in these compartments by NADPH production. PMID- 11482376 TI - Calcium handling by the cat carotid body--a pyroantimonate study. AB - Subcellular regulation mechanisms of calcium concentrations related to oxygen sensing in the carotid body are unclear. In the present study, we investigated the ultrastructural distribution patterns of calcium in carotid body cells and its changes evoked by hypoxia. Carotid bodies were dissected from anesthetized cats exposed in vivo to normoxic or acute hypoxic conditions. We used the oxalate pyroantimonate technique that yields an electron-opaque calcium precipitate. X ray microanalysis and appropriate controls confirmed the presence of calcium in the precipitate. Calcium precipitates were found in all types of cells in carotid body parenchyma: chemoreceptor cells, sustentacular cells, and nerve endings. In normoxic chemoreceptor cells, the precipitate was localized in dense core vesicles, mitochondria, and nuclei, but rarely in the cytoplasm. The most apparent effect of hypoxia was disappearance of the precipitate from dense core vesicles and was associated with its appearance in the cytoplasm. The amount of precipitate throughout the carotid body parenchyma was decreased overall due to hypoxia. These results indicate the involvement of subcellular calcium trafficking in hypoxia-sensing in the carotid body. The redistribution pattern of granular calcium deposits from organelles to the cytoplasm of chemoreceptor cells agrees with biochemical data of calcium release from intracellular stores during hypoxia. PMID- 11482377 TI - Carbohydrate histochemistry of lamb duodenum. AB - In order to elucidate the carbohydrate profile of the mucosa of lamb duodenum, conventional histochemical methods and a panel of 7 labelled lectins were used. In some cases, treatment with sialidase preceeded lectin staining. Carbohydrate histochemistry revealed the presence of sugar residues in the brush border of enterocytes, goblet cells and duodenal glands. All sites contained neutral and acid glycoconjugates. The presence of sulphomucins in goblet and duodenal gland cells was age-dependent. Enterocytes and duodenal gland cells contained abundant amounts of oligosaccharides with terminal sialic acid-galactosyl(beta1 --> 3)N acetylgalactosamine, whereas goblet cells contained the penultimate N acetylgalactosamine residue linked to sialic acid. These findings were not age dependent, whereas scarce amounts of fucose were found in all sites especially in young animals. The findings obtained in the present study serve as a basis for future pathological studies in lamb and sheep. PMID- 11482378 TI - Immunocytochemical localization of nucleophosmin and RH-II/Gu protein in nucleoli of HeLa cells after treatment with actinomycin D. AB - HeLa cells were studied with the use of light microscopical immunocytochemistry to obtain more information on the nucleolar localization of nucleophosmin (B23 protein) and RH-II/Gu protein. In control cells these proteins are colocalized. After structural rearrangement of nucleoli induced by actinomycin D, nucleophosmin was present in the nucleolar periphery and nucleolar peripheral caps. In contrast, RH-II/Gu protein was observed mainly in central regions of nucleoli as a compact body. However, in some nucleoli the proteins colocalized in a small portion of the peripheral nucleolar cap. The proteins were also colocalized in micronucleoli but occasionally RH-II/Gu protein was absent or both proteins were not present. PMID- 11482379 TI - Early histological changes of ileal mucosa after augmentation cystoplasty. AB - Segments of bowel are used routinely for transplantation in various pathological conditions such as contracted bladders or poorly compliant neuropathic bladders. However, little is known how these intestinal segments adopt to a toxic environment caused by urine. Therefore, the present study was performed to determine early histological changes of ileal mucosa after augmentation cystoplasty. Seven patients with augmentation cystoplasty underwent random cold cup biopsies of ileal segments after a mean period of 14.4 months after cystoplasty and morphological changes were evaluated using light microscopy and transmission and scanning electron microscopy. Most pronounced features were varying degrees of villous atrophy, increased numbers of Paneth and goblet cells. Severity of atrophic villous changes were not related to the length of the interval between surgery and endoscopic biopsy. These findings may be explained as adaptations of bowel tissue to counteract noxious effects of urine and to maintain its epithelial function in the bladder. PMID- 11482380 TI - Correlation between aerosols, deposits and weathering crusts on ancient marbles. AB - The structure and physicochemical characteristics of weathered surfaces of marble in industrial environments can be attributed to the interconnected evolution of the processes taking place at the atmospheric environment--marble interface. The present work is an attempt to correlate the aerosols of a heavily polluted atmosphere with the different weathering patterns observed on marble surfaces. Energy Dispersive X-Ray Fluorescence, X-Ray Diffraction, Porosimetry, Atomic Absorption, Atomic Emission Spectrometry, Ion Chromatography, Optical Microscopy and Scanning Electron Microscopy results were used together in principal component and discriminant analysis. These analyses were performed on forty six samples of aerosols and eighteen samples of crusts. Other parameters like orientation of the weathered surface, exposure to rainfall, presence of recrystalised calcite and gypsum were also used. The samples were collected from the archaeological site of the Sanctuary of Demeter, located near Eleusis (west of Athens, Greece), where a great number of industries (mainly metallurgical and chemical) cause several environmental problems. The results provide invaluable information on the nature of marble surface decay. The elements determining the chemical composition of the coarse airborne particles are Ca, Si, S, Br and Cl. The presence of gypsum is strongly related to black crusts or loose deposits. Y, Mn and rain exposure, are correlated with each other and related to black-gray crusts. Ca and Sr are correlated with washed-out surfaces. Finally discriminant analysis is proved to be a powerful tool in prediction of the type of decay that will be occur on a marble surface, given the composition and type of the polluted atmosphere. PMID- 11482381 TI - Nutrient stimulation of sulfolane biodegradation in a contaminated soil from a sour natural gas plant and in a pristine soil. AB - Natural gas in western Canada can contain up to 35% H2S. The Sulfinol process for sour gas treatment makes use of sulfolane and an amine to remove H2S and other sour components from natural gas. Sulfolane has leached into groundwaters at sour gas treatment plant sites, and poses a risk for off-site contamination. Sulfolane biodegradation was monitored in shake-flask cultures and air-sparged microcosms inoculated with uncontaminated topsoil or with sulfolane contaminated soil obtained near a Sulfinol process building at a sour gas treatment facility in western Canada. For both soils, supplementation with a source of fixed nitrogen stimulated sulfolane biodegradation. Topsoil cultures and microcosms were only slightly affected by the addition of phosphate. Contaminated soil microcosms and cultures were stimulated by phosphate addition, but not to the same degree as by the addition of nitrogen. For these cultures and microcosms, amendment with both fixed nitrogen and phosphate produced an additive effect. It was possible to predict the nutrient requirements of air-sparged microcosms inoculated with each soil type using shake-flask cultures. Shake-flask cultures require less time and effort and fewer materials than the more complex air-sparged soil microcosms, and will be useful for large-scale experiments to predict the nutrient supplements required for bioremediation of sulfolane-contaminated sites. PMID- 11482382 TI - Reduction of indicator organisms during biological solubilization of metals: a pilot study. AB - Laboratory and pilot scale studies on the biological solubilization of metals from undigested raw sludge were carried out using elemental sulphur as the energy source. Metals (Cu, Zn, Ni and Cd), pH, sulfate and indicator bacteria [total coliforms (TC) faecal coliforms (FC) and faecal streptococci (FS)] were used to evaluate the system effectiveness. This paper does not report the detailed design and operational data of the pilot plant (available elsewhere [1]) but focuses on the biological tests carried out at the Main Wastewater Treatment Plant in Toronto from July to December, 1997. Since the indicator bacteria are injured from exposure to low pH and potentially toxic metal concentrations, improved alternative methods for their enumeration and their ability to repair acid-metal induced injury in a resuscitation medium (CASO tryptone soya broth) were also investigated. Resuscitation was not effective in repairing cells injured during bacterial leaching. The MPN technique (using lauryl tryptose broth) for enumerating indicator bacteria (total coliforms) gave higher counts and was therefore superior to the spread plate technique (using m-Endo agar) in recovering bacteria from acidic leached sludge (biosolids). However, the coliform bacterial counts from raw sludge were similar by the two methods. This study indicated that the biological solubilization process could significantly reduce the pathogenic indicators. Concentrations of TC, FC and FS in the leached biosolids from the solubilization tank were lower than concentrations in the raw sludge by 4 to 6 orders of magnitude and the finished product (biosolids) met US. EPA requirements for pathogen and metal concentrations for Class A biosolids to be used on agricultural land. PMID- 11482383 TI - Variations of landfill leachate's properties in conjunction with the treatment process. AB - A study was done on the variations of water quality parameters, in conjunction with the processing steps, installed for the treatment of leachate from a sanitary landfill. The leachate was primarily subjected to biological treatment, composed of anaerobic digestion, aerobic treatment, and rotating biological contact in series. The effluent from the rotating biological contact process was further treated by combination of flocculation-sedimentation and adsorption processes. Finally, it was polished by the reverse osmosis process before discharge. The quality of raw leachate, of influents to and of effluents from the reverse osmosis process was assessed and compared. To determine size--dependent behavior of organic materials, analyses were also made for the fractionated samples through ultrafiltration. The overall treatment efficiency was about 98% for removal of organic materials. Most of the organic materials were in the low molecular weight range. In particular, about 95% of the biological oxygen demand was found to be exerted by the fraction of organic material of which cut -off molecular weight was less than 500. The analysis of molecular weight distribution confirmed that the organic materials resistant to the removal by the treatment process were humic substances, of which cut off molecular weight was greater than 500. The reverse osmosis process showed a high efficiency for removal of high molecular weight constituents. However, the final effluent showed an unexpectedly high oxygen demand. PMID- 11482384 TI - Phonological studies of the new gas-induced agitated reactor using computational fluid dynamics. AB - An ozone-induced agitated reactor has been found to be very effective in degrading industrial wastewater. However, the cost of the ozone generation as well as its short residence time in reactors has restricted its application in a commercial scale. An innovated gas-induced draft tube installed inside a conventional agitated reactor was proved to effectively retain the ozone in a reactor. The setup was demonstrated to significantly promote the ozone utilization rate up to 96% from the conventional rate of 60% above the onset speed. This work investigates the mixing mechanism of an innovated gas-induced reactor for the future scale-up design by using the technique of computational fluid dynamics. A three-dimensional flow model was proposed to compute the liquid gas free surface as well as the flow patterns inside the reactor. The turbulent effects generated by two 45 degrees pitch-blade turbines were considered and the two phases mixing phenomena were also manipulated by the Eulerian-Eulerian techniques. The consistency of the free surface profiles and the fluid flow patterns proved a good agreement between computational results and the experimental observation. PMID- 11482385 TI - Improvement of water treatment performance by using polyamine flocculants. AB - In this study, polyamine flocculants were synthesized and applied to Nak-dong river raw water in Korea to examine their efficiency in reducing turbidity, total organic carbon (TOC) and UV254. Synthesized polyamines were effective as flocculants for water treatment and the addition of organic polymer caused a reduction of 50-80% of the consumption of polyaluminium chloride (PAC). The effects of polyamine on the removal of turbidity, TOC and UV254 were investigated via both jar and pilot tests. The adsorption and separation mechanisms for the removal of turbidity and TOC by using the polymer flocculants were also observed. PMID- 11482386 TI - Biological sulfide oxidation in a fluidized bed reactor. AB - Feasibility of a laboratory scale fluidized bed process for biological sulfide oxidation to elemental sulfur and the formation of well-settleable sulfur sludge is demonstrated. Sulfide oxidation strongly depends upon oxygen concentration, sulfide loading rate and upflow velocity. At reactor dissolved oxygen concentrations (DOr) higher than 0.1 mg l(-1), sulfate was the main product of sulfide oxidation Upon increasing the sulfide loading rate, the sulfate production rate decreased as sulfide oxidation to sulfur showed marked increase. Low formation of sulfate could mean that sulfide was inhibitory to sulfate producing bacteria or that conversion of sulfide to sulfur was more favorable than sulfate production. Sulfide conversions higher than 90% were obtained at sulfide loading rates of 0.13-1.6 kgS mr(-3) d(-1). At DOr less than 0.1 mg l( 1), sulfur was the major end product of the sulfide oxidation. Upflow velocity in the range of 16-26 m h(-1) and sulfide loading rate of 0.9-1.6 kgS mr(-3) d(-1) were necessary for generation of biogranules containing 65-76% of elemental sulfur. The elemental sulfur production of 76% was obtained at upflow velocity of 17 m h(-1) with sulfide loading rate up to 1.6 kgS mr(3)d(-1). Morphological examination of the biogranules showed elemental sulfur deposition in the sludge granule and outside the bacterial cells. PMID- 11482387 TI - Carbonaceous oxidation using a new vertically moving biofilm system. AB - This laboratory study consisted of the construction and testing of a new experimental biofilm system for the carbonaceous oxidation of a synthetic wastewater. In the system, a biofilm cuboid module with a high surface area was vertically and repeatedly immersed in and lifted out of the wastewater in a reactor. The performance of two different biofilm modules were compared: one module was constructed from crossflow corrugated plastic sheets with a specific surface area of 410 m2m(-3) and had a calculated surface area of 6.24 m2; the other consisted of honeycombed plastic with hexagonal vertical columns and an estimated surface area of about 2.8 m2. Filtered chemical oxygen demand (COD) removal rates per bulk fluid volume for the corrugated and honeycombed modules of 7.2 kg COD m(-3) d(-1) and 7.6 kg COD m(-3) d(-1) were respectively achieved and these rates compare favourably with other wastewater treatment systems. The new biofilm system was simple to construct and operate, and was very effective in removing biodegradable COD from the synthetic wastewater. The system offers potential for reduced reactor volumes, energy saving, low solids production and easy solids removal. PMID- 11482388 TI - Screening of four species of detritivorous (humus-former) earthworms for sustainable vermicomposting of paper waste. AB - Four specices of detritivorous (humus-former) earthworms were tested for their ability to vermicompost paper waste blended with cowdung in 6:1 (w/w) ratio. The anecic Lampito mauritii, Kinberg and the epigeic Eudrilus eugeniae, Kinberg were the most effective of the four species employed; 20 animals of each of these species generated castings amounting to about 52% of the feed mass (75 g) per fortnight. The performance of these two species was followed by the anecic Drawida willsi, Michaelsen and the epigeic Perionyx excavatus, Perrier; they achieved approximately 46% vermiconversion in comparable settings. The vermireactors were sustainable as the animals have remained consistently healthy and reproductive over a period of six months, and are continuing to remain so, turning in a steadily rising vermicast output. During this period E. eugeniae have grown to 2.3 times their original weight while the other three species have more than trebled their weights. The studies establish the feasibility of vermicomposting as a viable process for the gainful utilization of paper waste in an environmentally clean manner. They also indicate that all the four species of the worms screened by us are suitable for the process, with L. mauritii and E. eugeniae a shade more efficient than the other two species. PMID- 11482389 TI - Chemical characterization of PM2.5 aerosols in Athens-Greece. AB - Atmospheric aerosols, PM2.5 were simultaneously collected in the center of Athens and in a semi-urban area of the Athens basin, using the Harvard Impactor (HD) system, from March 1995 to March 1996. 224 24-hr samples were collected. Chemical analysis of the filter samples was performed using ion chromatography (Cl-, SO4(2 ); NO3-, Na+, K+, NH4+, Mg2+, Ca2+). In addition aerosol addity was measured using a semi-micro electrode. No significant differences in chemical composition of particles were observed between the two sampling sites. At the city center the annual-average of non-sea-salt sulfate concentration was 85 nmoles m(-3). Also the annual mean values of chloride, ammonium and sodium concentrations were 48, 88 and 71 nmoles m(-3) respectively. Lower concentrations were observed for the rest of the ions. Aerosol acidity was higher at the city center, 18 nmoles m(-3), compared to that observed at the semi-urban site, 14 nmoles m(-3). Species concentrations were examined by season. The mean monthly concentrations of Cr, NO3-, Ca2+ and H+ were higher in the winter. In contrast those of Mg2+ Na+ and K+, were higher in the summer and the spring, respectively. The concentrations of SO4(-2)and NH4+ ions did not exhibit a seasonal pattern. Sulfate and ammonium ions were the predominant ionic species and their ionic ratio ranged between those of ammonium sulfate and letovicite. PMID- 11482390 TI - Biological treatment of fishery washing water using Bacillus sphaericus coupled with production of spores that are toxic to mosquito larvae. AB - Research was undertaken to investigate the treatment of fishery washing water using Bacillus sphaericus, and to recover the spores for subsequent use as bioinsecticide to control the population of mosquitoes. This treatment method could reduce pollution due to organic matter by decreasing the value of Chemical Oxygen Demand (COD) and Biological Oxygen Demand (BOD) by about 85% and 92%, respectively. The maximum concentration of spores (83.3 x 10(7) spores ml(-1)) using normal concentration of filtered fishery washing water was only about 27% lower than that obtained in fermentation using 0.25% (w/v) yeast extract. The larvicidal activity of the spores produced in fermentation using fishery washing water to Culex quinquefaciatus, as measured by LD50 after 48 h, was almost the same as the larvicidal activity of spores obtained from fermentation using yeast extract. PMID- 11482391 TI - The effects of the flow pattern on organic oxidation and nitrification in aerated submerged biofilters. AB - Previous research has shown that a submerged biofilter is effective in removing organic matter and total organic nitrogen. Upflow and downflow biofilters were set up to investigate the different oxidative efficiencies on organic matter and ammonia nitrogen of different biofilter positions and to compare the biological activities of the upflow and downflow patterns. A comparison of the operational characteristics of the two types of biofilters is discussed. The main COD removal zone is 0-20 cm in the upflow biofilter, but in the downflow biofilter it occurs in the submerged zone. The nitrification process is dependent on the HRT and the residual COD. In the downflow biofilter, the efficiency of the nitrification increases with the HRT. The growth of nitrifying bacteria has a tendency to take place in higher zones in the downflow biofilter, especially for a longer HRT. Batch tests measured the biological activity of heterotrophs and nitrifying autotrophs with decomposition rates. In the upflow biofilter, the nitrification rates increased while the COD oxidative rates decreased with the height of the biofilter, so the effect of the HRT on the activity of the nitrifying autotrophs is significant. However, in the down biofilter, the activity of the nitrifying autotrophs was similar at different heights in the same HRT runs with the maximum nitrifying autotrophs activity being at an HRT of 6 hours. PMID- 11482392 TI - Effects of flow velocity and paving material, on nitrogen transformations, in shallow open channels. AB - A study was conducted to investigate the effects of the flow velocity and the paving material on the nitrogen conversion rates in shallow open channels. Equal amounts of hollow holed and plain balls, all with a 3.5-cm diameter, was fastened to the channel bottom. Then the nitrogen conversion rates were taken at various velocities and compared. The inner and outer surfaces of the holed ball channel were about 1.5 times than of the plain ball channel, but the attached biofilm mass was found to be about 3.55 times greater. The optimal nitrogen conversion rates were found to be higher in the holed ball channel. A suitable velocity was determined which would enhance the optimal nitrogen conversion rates. The choice of velocity and the biofilm carrier to enhance or predict the nitrogen transformations in shallow channels, should be considered in model development. PMID- 11482393 TI - Kinetics of color removal by adsorption from water using activated clay. AB - Kinetics and mechanism of adsorption of one commercial basic dye (BB69) and one direct dye (DR227) from aqueous solutions using day were studied in the temperature range 15-75 degrees C. The clay was activated by treatment with sulfuric add. Two simplified kinetic models, pseudo first- and second-order equations, were tested. It was shown that the adsorption of both dyes could be well described by the pseudo second-order equation. Rate constants of the best fit model were calculated, and the related apparent activation energies were also evaluated and discussed. PMID- 11482394 TI - An improved method for determination of heavy metal bioavailability in contaminated soil. AB - Bioavailability of heavy metal in contaminated soil is investigated. A general diffusion model for determining the heavy metal bioavailability in soil has been developed The bioavailability predictions based on the present model were more accurate than those based on a previous model. Experimental results obtained using cadmium-, copper-, zinc- and lead-contaminated soils were employed for model verification. The effects of soil pH, initial heavy metal concentration, temperature and soil type on the effective diffusion coefficient or bioavailability index were also examined experimentally. The theoretical model and experimental procedure proposed in the present study provide a convenient means for the determination of heavy metal or inorganic ion bioavailability in contaminated soil. PMID- 11482395 TI - Nonequilibrium mass transfer of multi-component NAPL in a soil column venting. AB - This study was conducted to investigate the mass transfer behavior for a multi component system in the soil venting process. Soil venting experiments were conducted using gasoline-contaminated soil and models of local equilibrium assumption (LEA) and the first-order kinetic approach were used to descnbe the gasoline volatilization process. However, the focus was on the application of the kinetic model. In both models, thirteen major components of gasoline were selected for the model input and the rest of the gasoline was divided into 11 groups based on the retention times in the gas chromatography of gasoline. The LEA model had the tendency of underestimating the gas concentration at the initial phase and overestimating at the later phase due to the different volatilities of multi-components. n the kinetic model, the estimation of mass transfer coefficient values were carried out by adopting the relationship developed in the single-components system and choosing the appropriate modified Sherwood number. This method resulted in the good agreement with the simulation and the experimental results. PMID- 11482396 TI - Polybrominated diphenyl ether (PBDE) flame retardants. AB - Polybrominated diphenyl ether, PBDE, flame retardants are now a world-wide pollution problem reaching even remote areas. They have been found to bioaccumulate and there are concerns over the health effects of exposure to PBDEs, they also have potential endocrine disrupting properties. They are lipophilic compounds so are easily removed from the aqueous environment and are predicted to sorb onto sediments and particulate matter or to fatty tissue, aiding their distribution throughout the environment. PBDEs are structurally similar to PCBs and DDT and, therefore, their chemical properties, persistence and distribution in the environment follow similar patterns. Concentrations of PBDEs found in environmental samples are now higher than those of PCBs. Evidence to date demonstrates that PBDEs are a growing problem in the environment and concern over their fate and effects is warranted. The manufacture of reactive and additive flame retardants is briefly discussed and their fate and behaviour in the environment is assessed. PBDE toxicology is reviewed and methods of analysis are evaluated. PMID- 11482397 TI - A systemic biokinetic model for polonium. AB - Although the biokinetics of polonium has been studied extensively, interpretation of the data is complicated by potential differences with species and route of exposure and the questionable reliability of much of the reported excretion data for man. A study was undertaken to identify the data that are most likely to represent the typical behavior of polonium and apply those data to construct an improved, physiologically realistic systemic biokinetic model for polonium in man. Such a model is needed for interpretation of urinary excretion data for workers exposed to 210Po and reconstruction of the radiation doses received by those workers. This paper reviews the database on the biokinetics of polonium and describes a new systemic biokinetic model for polonium in man. PMID- 11482398 TI - Simultaneous absorption of SO2 and NO from flue gas with KMnO4/NaOH solutions. AB - The wet scrubbing combined SOx/NOx removal system is an advanced air pollution control device. This study attempts to understand the absorption kinetics in the system. The absorption of diluted SO2 and simultaneous absorption of diluted SO2 and NO, as occurs in flue gases, in a stirred tank reactor with KMnO4/NaOH solutions were carried out at 50 degrees C. The liquid-side and gas-side mass transfer coefficients of the system were determined. The results indicate that the absorption of SO2 is close to completely gas-film controlled where the NaOH concentration is greater than 0.1 M or the KMnO4 concentration is greater than 0.05 M. The increasing gas flow rate has a positive effect on the absorption rate of SO2. The existence of O2 has no significant effect on the absorption rate of SO2. Adding SO2 would decrease the absorption rate of NO; however, the addition of NO has no effect on the absorption rate of SO2. PMID- 11482399 TI - OECD Environment Ministers adopt an environmental strategy for the next decade. Paris, 16 May 2001. PMID- 11482400 TI - Lead contamination in tap water of households with children in Lower Saxony, Germany. AB - Lead has numerous acute and chronic adverse effects on human beings. This is especially true for infants and children. The main path of lead ingestion in children can be different according to housing and living situation. The intake of lead through drinking water is commonly due to metal corrosion. The users plumbing can be an important factor. In recent years, many lead pipes in Germany have been replaced by pipes made of an alternative material. The aim of this study is to assess the present state of drinking water contamination and the resulting exposure of infants to lead. For this purpose mothers of new-born babies were offered a free examination of their drinking water. After a written declaration of consent had been obtained and after the infant in question had reached an age of 3 months, a stagnation sample of cold tap-water after overnight stagnation together with a random daytime sample was obtained from the family. The collected samples were analysed by atomic absorption spectrometry for their lead concentration. In total, 1485 samples from households were collected. Of the 1434 stagnation samples, 3.1% had lead concentrations greater than 0.01 mg/l (recommended limit of the WHO) and 0.6% had concentrations above the limit of the German drinking water regulation (0.04 mg/l). The values for the 1474 random daytime samples were 2.1% above 0.01 mg/l and 0.2% greater than 0.04 mg/l, respectively. By region, the areas Bovenden, Friedland, Duderstadt, Northeim and Rosdorf were particularly affected. The highest measured concentrations of lead in the stagnation samples were 0.11 mg/l and 0.15 mg/l in the random daytime samples, respectively. PMID- 11482401 TI - Divalent metal accumulation in freshwater bivalves: an inverse relationship with metal phosphate solubility. AB - Whole soft tissue concentrations of Mn, Co, Ni, Cu, Zn, Pb, Cd and U were measured in two species of freshwater (unionid) bivalves (Hyridella depressa and Velesunio ambiguus) from a minimally polluted site in the Hawkesbury-Nepean River, south-eastern Australia. Although the mean concentrations of metals in the tissue were similar for each bivalve species, their patterns of accumulation were dissimilar. For each metal, positive linear relationships between tissue concentration and shell length (r2 = 0.37-0.77; P < or = 0.001) and tissue dry weight (r2 = 0.29-0.51; P < or = 0.01) were found in H. depressa, but not in V. ambiguus. However, for both species, positive linear relationships were found between the tissue concentration of each divalent metal and Ca tissue concentration (r2 = 0.59-0.97; P < or = 0.001). For both bivalve species, the normalised rates of accumulation of the metals relative to increasing Ca concentration and/or size, were U approximately = Cd > or = Pb > or = Mn > Co > or = Zn > Cu > Ni. The differential rates of accumulation of divalent metals are interpreted as being predominantly governed by their varying loss rates, which are controlled by the differing solubilities (log Ksp values) of the metals in the phosphatic extracellular granules, the demonstrated major sites of metal deposition in the tissue of H. depressa and V. ambiguus. The rates of accumulation of Mn, Co, Zn, Cu and Ni were linearly and inversely related (r2 = 0.91-0.97; P < or = 0.001) to their solubilities as hydrogen phosphates, a finding consistent with the bioaccumulation model previously developed for the alkaline-earth metals. However, for U, Cd and Pb, this linear inverse relationship did not continue to hold, i.e. their rates of accumulation did not increase with decreasing solubility. However, these results are still consistent with the model if U, Cd and Pb are so insoluble in the granules of H. depressa and V. ambiguus over their lifetime (up to approx. 50 years) that there is effectively no loss of these metals, and hence, no differential between their rates of accumulation. The present results reaffirm the use of Ca tissue concentration to predict the tissue concentrations of other divalent metals by explaining up to 94 and 97% of the variability between individual bivalves of H. depressa and V. ambiguus, respectively. The use of Ca tissue concentration to effectively minimise the inherent variability between individuals in their metal tissue improves the ability of an investigator to discern smaller spatial and/or temporal differences in the metal tissue concentrations of these bivalves, and thus to detect metal pollution. PMID- 11482402 TI - The performance of two lichen species as bioaccumulators of trace metals. AB - The concentrations of 16 metals were measured in peripheral and central parts of the foliose epiphytic lichens Parmelia caperata and Xanthoria parietina collected in north-east Italy. Relevant intra- and interspecific variability was shown by multivariate analysis (classification and ordination) of the matrix of metals and samples. The central, older parts of the thalli contained significantly higher amounts of most metals in both species. Significative differences were also found between peripheral parts of the two species, with higher concentrations of Fe and Al in Xanthoria, and Cd and Zn in Parmelia. An inter-species comparison of several hundred Italian measurements confirmed the higher affinity of Parmelia for Cd and Zn, but not that of Xanthoria for Fe and Al. In order to enhance data quality in biomonitoring studies, it is suggested to analyse only peripheral parts of the lichens, and to avoid the joint use of Parmelia and Xanthoria when monitoring Cd and Zn. PMID- 11482403 TI - Speciation patterns of heavy metals in tropical estuarine anoxic and oxidized sediments by different sequential extraction schemes. AB - The speciation patterns of Cu, Cd, Zn, Pb, Fe and Mn in sediment samples under anoxic and oxidized conditions were investigated using three-stage, four-stage and five-stage sequential extraction schemes. All the extraction schemes identify the non-residual metal among three basic operationally-defined host fractions, namely, exchangeable, reducible and organic/sulfide bound. The anoxic sediment samples were found to have been oxidized during the extraction stage for the reducible fraction under the three-stage and four-stage schemes and the moderately reducible fraction under the five-stage scheme despite the maintenance of an oxygen-free environment. This artifact has resulted in an over representation of the reducible fraction and an under-representation of the organic/sulfide fraction in the heavy metal speciation patterns of anoxic sediment samples. For Cd, Zn and Pb which had > 70% associated with the acid volatile sulfide in the organic/sulfide fraction, this artifact has resulted in the observation of a decrease in the reducible fraction and, in some cases, an increase in the organic/sulfide fraction upon oxidation of the anoxic sediment samples. PMID- 11482404 TI - How effective has been the reduction of SO2 emissions on the effect of acid rain on ecosystems? AB - The paper attempts to assess the effectiveness of the reduction of SO2 emissions over recent years for protection of ecosystems in both Europe and eastern USA by analysis of temporal changes in the acidifying potential (AP) of wet deposition defined as [SO4(2-)] - ([Ca2+] + [Mg2+]). Spatial and temporal patterns of acidifying potential were studied. The main result is that there is no statistical evidence for a trend towards improvement in the acidifying potential, because there have been declines in Ca precipitation. This suggests that the reduction of SO2 emissions has not been effective as an abatement strategy. PMID- 11482405 TI - Is the Negro River Basin (Amazon) impacted by naturally occurring mercury? AB - In order to investigate the major sources and cycling of mercury in the Negro River Basin (Amazon), total metal measurements were carried out in soils, in river and lake waters, in the atmosphere, and in bulk precipitation during the period 1995 throughout 1998. Median values of 1.3 ng m(-3) in the atmosphere, 172 microg kg(-1) in soils, 4.6-7.5 ng l(-1) in three different lakes, 4.5 ng l(-1) in 17 different Negro River tributaries and 20.3 microg m(-2) year(-1) in bulk precipitation were found. Mercury concentrations in rivers and lakes waters, as well as in soils and bulk precipitation were high, considering the scarcity of anthropogenic point sources in the region. Mercury leaching from soil, the largest regional reservoir of this metal, was considered to be the major pathway to mercury enrichment in the region. PMID- 11482406 TI - The kinetics of catalytic incineration of styrene over a MnO/Fe2O3 catalyst. AB - Catalytic incineration is one of the cost-effective technologies to deal with unwanted volatile organic compounds (VOCs). Catalytic incineration of styrene over a MnO/Fe2O3 catalyst was carried out in a bench scale catalytic incinerator. Three kinetic models, the power-rate law, the Mars and van Krevelen model and the Langmuir-Hinshelwood model were used to analyze the results. A differential reactor design was used for best fit of kinetic models in this study. The results show that the Langmuir-Hinshelwood model may be feasible to describe the catalytic incineration of styrene. This suggests that the chemical adsorption of either O2 molecules or O atoms is important in the process of catalytic incineration of styrene. PMID- 11482407 TI - A comparative assessment of Boise, Idaho, ambient air fine particle samples using the plate and microsuspension Salmonella mutagenicity assays. AB - The primary objective of this study is to characterize the genotoxic potential of the ambient air aerosols collected within an air shed impacted primarily by wood smoke and automotive emissions. The study also examines the relative merits of a microsuspension assay and the standard plate assay for monitoring the presence of airborne particle-bound mutagens. Wintertime ambient air particulate samples collected from Boise, Idaho, USA, were shown to contain extractable organic matter that is mutagenic in the Salmonella typhimurium microsuspension and plate incorporation assays. Differences in the results from the primary sites, auxiliary sites and the background site demonstrate that the particle-bound mutagens are not evenly distributed within the air shed and are more associated with the location of sampling than with the time of sampling or the type of bioassay used to evaluate the samples. This study also demonstrates that the bioassay protocol used in such studies should depend upon the characteristics of the air shed's mutagens and the purpose of the study. For example, the microsuspension assay gave somewhat more variable results between samples but was approximately threefold more sensitive than the plate assay. When strain TA98 was used in the microsuspension assay, the mutagenic response was greater without an exogenous activation system. The reverse was true for the plate assay in which the use of an exogenous activation system increased the mutagenicity response. TA100 in the microsuspension assay provided results comparable to those with TA98. This is important because TA100 can also be used to bioassay semivolatile and volatile organics associated with ambient air mutagenicity. This, in turn, allows a comparison of the mutagenicity of organics collected by differing methods due to their volatility. Future studies should be directed toward correlation of mutagenicity results with other analytical results in order to further develop methods for better characterization of the genotoxicity of ambient air. PMID- 11482408 TI - Audit in general practice: how much and how complete? Frequency of audit in general practice. AB - The aim of this study was to determine the frequency of audit and the proportion completed in a group of practices. Data obtained by interviewing a member of each practice and inspecting practice records were independently coded by researchers. Practices initiated an average of 3.5 (range 1-7) audits in 2 years, of which an average of 0.9 (range 0-3) were completed. Ten of 16 completed audits were externally funded or facilitated. Few audits are completed in general practice and practices require continuing support for audit. PMID- 11482409 TI - General practitioners' reactions to non-compliant patients. AB - OBJECTIVE: To examine how general practitioners (GPs) respond to patients who are non-compliant with medical advice and who doctors believe act irresponsibly towards their health. DESIGN: Quantitative analysis of responses to questionnaire with case histories. SETTING AND SUBJECTS: 93 questionnaires completed by a random stratified sample of Norwegian GPs. MAIN OUTCOME MEASURES: Scores relating to GPs' feelings and choice of main and sub-strategies for further treatment of patients. RESULTS: The respondents typically felt discouraged or unaffected by non-compliant patients, younger doctors more often felt helpless while older ones were more content, and female doctors more often than male doctors felt irritated or angry. The main strategy preferred was to give the patient a new appointment. The young, the inexperienced, and females tended more often to ask a colleague for advice or refer to a specialist. Patient-centred sub-strategies were generally preferred, especially by younger doctors. CONCLUSION: GPs' feelings towards and strategies for dealing with non-compliant patients vary, and the doctor's age, sex and clinical experience are central variables. PMID- 11482410 TI - Headache and associations with lifestyle among pupils in senior level elementary school. AB - OBJECTIVE: To investigate the prevalence of headache and its association with lifestyle among schoolchildren and to test an intervention programme. DESIGN: A cross-sectional survey of all pupils in the school who were asked to complete a questionnaire about headache and lifestyle factors. SETTING: School health care at an elementary school in Sweden. SUBJECTS: All 344 pupils aged 13-16 years, 170 girls and 174 boys, at senior level answered the questionnaire and were included in the study. MAIN OUTCOME MEASURES: Headache occurring at least once a week. RESULTS: Twenty-two percent had headache at least once a week. The problem was more common among girls than among boys; OR 1.6 (95%CI 1.3-2.1). In girls, headache was associated with smoking; OR 6.6 (CI 1.2-35.5), going to bed later than 11 p.m.; OR 4.4 (CI 1.1-18.0), headache in parents; OR 2.0 (CI 1.0-4.2), and few sports activities; OR 3.0 (CI 1.2-7.5). The only corresponding association in boys was with smoking; OR 12.0 (CI 1.5-101). Headache improved in pupils participating in the programme. CONCLUSIONS: Headache is a common health problem in schoolchildren of both sexes at senior level and shows strong associations with lifestyle factors in girls. Intervention seems to be efficient. PMID- 11482411 TI - Five-year mortality in men and women with atrial fibrillation. AB - OBJECTIVE: Five-year mortality in men and women with atrial fibrillation (AF). DESIGN: A follow-up of the medical records of patients with AF registered on September 30, 1993. SETTING: One community health centre in Stockholm County. SUBJECTS: 129 patients (76 men and 53 women). MAIN OUTCOME MEASURES: Observed and expected, age- and sex-standardised, 5-year mortality rates. Significant risk factors by multiple logistic regression. RESULTS: Women had a higher mean age (77.5 vs 72.8 years) and more often suffered from heart failure and hypothyreosis. Five-year mortality rate for men was 30% vs expected 27% in Sweden and 24% in the community; for women it was 43% vs expected 29% (p <0.05) and 23% in the community (p < 0.001), i.e. an excess mortality of 49% and 88%, respectively. Significant factors predicting death by logistic regression among women were: age (odds ratio 1.39, p < 0.001), levothyroxine treatment (odds ratio 27.87, p < 0.05) and diabetes (odds ratio 20.75, p < 0.05). CONCLUSIONS: AF is related to an excess sex- and age-standardised, 5-year mortality in women but not in men, with levothyroxine treatment as one significant factor. PMID- 11482412 TI - Diagnostic evaluation of dementia in general practice in Denmark. A national survey. AB - OBJECTIVE: To examine GPs' self-reported basic diagnostic evaluation of dementia according to the recommendations in multidisciplinary consensus guidelines and to analyse explanatory factors for GP performance. DESIGN: Postal questionnaire study, spring 1998. SETTING: General practice in Denmark. SUBJECTS: All 3379 GPs in Denmark. RESULTS: The questionnaire was completed by 75.1%. According to our Diagnostic Evaluation Index, 47.2% of the GPs were classified as conducting a good basic diagnostic evaluation of dementia, and tended to have the following characteristics: they conducted regular follow-up consultations with demented patients (odds ratio (OR) 2.4), they were inclined to state that all patients with possible dementia should undergo diagnostic evaluation (OR 2.0), they considered that the GP should play the major role in diagnostic evaluation (OR 1.7) and they believed their methods were adequate identify dementia (OR 1.7). Gender, attitude to development of clinical guidelines and being able to refer patients to specialists were not influential. CONCLUSION: We conclude that the previously reported reluctant attitude of GPs is now more positive. PMID- 11482413 TI - Risk factors for respiratory tract infections in children aged 2-5 years. AB - OBJECTIVE: To study risk factors for respiratory tract infections (RTIs) in children aged 2-5 years. DESIGN: A questionnaire was used to chart the number of visits to doctors and the number of antibiotics courses because of RTIs during a 1-year period. The risk factors studied were age, sex, birth weight, breastfeeding, atopy, siblings, smoking and childcare. SETTING: The catchment area of Oxie health centre in southern Sweden. SUBJECTS: 190 pre-school children. MAIN OUTCOME MEASURES: Number of visits to doctors and number of antibiotics courses. RESULTS: The risk of having to consult a doctor, as well as the risk of receiving treatment with antibiotics, because of RTIs was greater among children at daycare centres than among children who were mostly at home (adjusted OR 2.78 (95% CI 1.34-5.78) and 2.73 (1.38-5.43), respectively). CONCLUSION: Attending a daycare centre is the most important risk factor for RTIs in children aged 2-5 years. PMID- 11482414 TI - Bacteriological findings in nasopharynx specimens from patients with a clinical diagnosis of acute sinusitis. AB - OBJECTIVE: To study results from bacteriological specimens from nasopharynx in patients with a clinical diagnosis of acute sinusitis in relation to CT findings. DESIGN: Prospective study. SETTING: Patients from general practice in Vestfold county, Norway. PATIENTS: 427 patients 15 years and older from two studies with a clinical diagnosis of acute sinusitis, and who were examined with coronal CT scans of the paranasal sinuses. Fluid level or total opacification of any sinus was taken as a hallmark of sinusitis. MAIN OUTCOME MEASURES: Bacteriological findings in nasopharynx specimens and proportions of various sinus pathogens in patients with and without sinusitis confirmed by CT. RESULTS: In the study, 252 patients had acute sinusitis and 175 patients did not. In the sinusitis groups, 27% of the patients had Streptococcus pneumonia, 12% had Staphylococcus aureus and 10% had Haemophilus influenzae in their nasopharynx specimens. Forty-five percent of the patients had normal nasal flora or no growth. The strains of Streptococcus pneumonia and Haemophilus influenzae showed high sensitivity to PcV, while the Moraxella strains were resistant to it. CONCLUSION: Streptococcus pneumoniae and Haemophilus influenzae were the most frequent sinus pathogens found in the nasopharynx specimens, and they were significantly more frequent in the group with confirmed sinusitis. The proportion of specimens with normal nasal flora or no growth was significantly higher in the non-sinusitis group. PMID- 11482415 TI - Is general practice effective? A systematic literature review. AB - OBJECTIVE: To find evidence of the effectiveness of physicians working in primary care. DESIGN: Systematic literature search in the Medline and Cochrane databases. MATERIAL: Out of 7223 titles found in the search, 45 studies, comparing, from different aspects, primary care with specialist care, were extracted. MAIN OUTCOME MEASURES: Health indicators, health care costs, quality of health care. RESULTS: Primary care contributed to improved public health, as expressed through different health parameters, and a lower utilisation of medical care leading to lower costs. Physicians working in primary care, in comparison with other specialists, took care of many diseases without loss of quality and often at lower cost. The organisation of primary care was important in respect of reimbursement by capitation, more group practices, higher personal continuity, and having generalists as primary care physicians. CONCLUSIONS: To compare the effectiveness of primary care and specialist care is a complex task and there are limitations in all studies. However, we have found evidence that increased accessibility to physicians working in primary care contributes to better health and lower total costs in the health care system. It is also clear that studies with evaluation of how to most effectively organise primary care are far too few. There is an extensive need for future research in this area, a suitable task for collaborative research between the Nordic countries. PMID- 11482416 TI - Report from University College London (UCL). PMID- 11482417 TI - "In really great danger..." The concept of risk in general practice. AB - The language of risk has many meanings, and in this article we demonstrate a discrepancy between individually perceived risk and the medical understanding of risk as understood and communicated by general practitioners (GPs). Risk is experienced and interpreted by people in a cultural context, i.e. the same objective risk can be perceived in many different ways and given a different meaning in daily life. GPs' evaluation of risk is made on the basis of our epidemiological knowledge, the medical culture of risk perception and the GP's personal experience and interpretation. The theoretical focus in the article is a synthesis of two theories: patient-centred general practice and theory based in anthropology about risk as culturally and socially constructed. We use empirical data from a qualitative study to illuminate the discussion. PMID- 11482418 TI - Patients, diagnoses and processes in general practice in the Nordic countries. An attempt to make data from computerised medical records available for comparable statistics. AB - OBJECTIVE: To try out a collection of a standard set of data from computerised medical records. DESIGN: Retrospective extraction of ordinary patient record information put into the computer by general practitioners. SETTING: Encounters in office hours in strategically selected practices or health centres in Denmark, Finland, Iceland, Norway and Sweden. SUBJECTS: 59 general practitioners and a total study population of 97475 persons. MAIN OUTCOME MEASURES: Proportions, crude and specific rates of encounters, diagnoses and processes. RESULTS: In a 4 week period there was a threefold difference in the office encounter rates between the participating sites in the Nordic countries. Gender and age patterns were similar despite these differences. An access to several different denominators revealed diverse patterns of referring to the specialist, prescribing, ordering blood tests, X-rays and physiotherapy. Data from computerised medical records agree well with earlier studies in the Nordic countries using other methods. CONCLUSIONS: This survey demonstrates that valid and reliable data for routine statistics are available from computerised medical records in general practice. The major obstacle extracting more epidemiological data from computerised medical records is caused by information in the databases not being uniquely linked to episodes of care. PMID- 11482419 TI - The diagnosis of cancer in the "roar" of potential cancer symptoms of patients in primary health care. Research by means of the computerised journal. AB - OBJECTIVE: To describe the diagnostic work-up pattern in primary health care, aiming, with as few diagnostic activities as possible, to identify a number of malignancies among patients presenting with various symptoms, where a malignancy may be a differential diagnosis. DESIGN: Survey of computerised journals. Diagnostic codes (ICD-9 system in primary health care) relating to signs, symptoms or diagnosis were selected where colorectal, pulmonary, breast and prostate malignancies might be differential diagnoses. All diagnostic actions were analysed. SUBJECTS: 6812 patients over 30 years of age from four health centres who were recorded for a total of 14455 selected diagnostic codes. RESULTS: The diagnostic actions resulted in 1426 X-ray or sonographic investigations, 340 endoscopies, 16203 haematology, clinical chemistry or microbiology tests and 667 referrals to specialists. Forty-nine malignancies were diagnosed at the primary health care centres, while 10 malignancies were classified as "missed". The frequency of faecal-occult blood tests performed was low while that of ESR and pulmonary X-ray examinations was high. CONCLUSION: The task for a GP identifying one or two undiagnosed malignancies per year of the four most common types among all the non-neoplastic ailments, and with as little diagnostic activity as possible, is a professional challenge to be scrutinised continuously. PMID- 11482420 TI - Specialists as consultants to GPs. Private sector services as an alternative way of organising consultant services in health care. AB - OBJECTIVE: To assess the effect on referral rate when GPs are given a better opportunity to send their patients to specialists for consultation. DESIGN: Intervention study with a control group. SETTING AND SUBJECTS: A 34-month experiment in the City of Turku. The experimental group consisted of 10 GPs working in municipal health centres with a list system and serving 23000 residents. As a control group, there were four GPs without a list system serving 10800 residents. OUTCOME MEASURES: GP visits and their referrals to specialists. RESULTS: The number of patient visits was higher among GPs in the experimental group than among those in the control group. During the experiment, the referral rate of GPs in the experimental group increased from 5.7% to 6.8%, but there was no change among the control GPs. Of all referrals to specialists, the share to the private sector increased from 5% to 35%, while at the same time the share to the hospital outpatient clinic decreased. CONCLUSIONS: An enhanced possibility to consult private specialists increases the number of referrals, but there are no consequent changes in the relative shares of the consulted specialties. PMID- 11482421 TI - The effect of an out-of-hours reform on attendance at casualty wards. The Danish example. AB - OBJECTIVE: A reorganisation of the out-of-hours general practice service in Denmark was launched in January 1992. The biggest changes were in a mandatory telephone triage staffed by GPs and the replacement of small rota systems with county-based health centres. We aimed to analyse the effect of this out-of-hours reform on the number of contacts with the casualty wards. DESIGN: A register based ecologic time-trend study of the mean number of annual contacts per inhabitant from 1988 to 1997. SETTING: The County of Aarhus. SUBJECTS: All 630000 inhabitants in the county. MAIN OUTCOME MEASURES: Mean number of annual contacts with casualty wards per inhabitant. Intercepts derived from two regression models. RESULTS: The mean number of contacts with casualty wards rose significantly during the whole period. Given this constant increase in contact rates, a regression model showed that the increase in the attendance rate with casualty wards after the reform was statistically insignificant. CONCLUSIONS: The decrease in the total number of contacts with the out-of-hours primary health care after the reform was not met by a corresponding increase in casualty ward contacts. A clear-cut significant increase in the use of casualty wards following the out-of-hours reform could not be demonstrated. PMID- 11482422 TI - Confidence and experience in emergency medicine procedures. Norwegian general practitioners. PMID- 11482423 TI - Substance use among gay and lesbian adolescents. AB - Surveys at a Massachusetts high school used multiple items to identify gay, lesbian, and bisexual teenagers. Students with consistent homosexual preferences had greatly elevated rates of substance use. Those who displayed a homosexual preference but answered less consistently also had somewhat higher rates of substance use but were closer to heterosexual youths than to more consistent homosexual youths. The differences in substance use were least for alcohol use and greater for "hard" drugs. PMID- 11482424 TI - Sex and sexual orientation: the effect of group membership on individuals' judgments about self and others' HIV risk. AB - No previous study has directly compared homosexual and heterosexual men and women's perceptions of HIV risk. In fact, empirical research focusing on homosexual women's perception of HIV risk is scarce. This paper, therefore, examines whether homosexual and heterosexual women and men (N = 60) make varied self and other (peer and non-peer) HIV risk judgments. The paper also examines the roles of motivational (health anxiety) and cognitive (HIV knowledge) factors in relation to HIV risk judgments. The results show that each group held different perceptions of risk for various "other" groups. Only homosexual men showed evidence for an optimistic bias, whereas homosexual women showed evidence of realistic perceptions. Both cognitive and motivational factors were shown to be associated with risk judgments for homosexual women and heterosexual men. Methodologically the use of Multidimensional scaling as an analytic strategy is recommended. PMID- 11482425 TI - Making Christabel: sexual transgression and its implications in Coleridge's "Christabel". AB - Even among critics who recognize the role of lesbianism in "Christabel," none consider the implications of lesbianism for the characters. Many readers describe Geraldine as a kind of supernatural power, a demon. But Geraldine's identity is far from clear, and we don't need to explain away the lesbian sexuality as demonically-inspired in order to understand the dynamics of the text. The poem accounts for its characters' interactions on what is a more visceral and less fantastic level, the implications of a lesbian act in the world of "Christabel." In "Making Christabel," I consider the protagonist's social and psychological stresses in terms of an encounter with lesbian sexuality in order to understand the poem's ambiguities: Geraldine's guilt, Christabel's pleasure, the manipulation of gender roles, and the dynamics between Christabel, Geraldine, and Sir Leoline. To this end, I look at how the "unnatural" sexuality between Christabel and Geraldine is marked by a reversal of gender expectations. Sexual transgression is suggested not simply by two women heading off to share one bed, but by the manipulation of gender roles on their way to and within the bedroom. I also read the homosocial bond be tween Sir Leoline and Sir Roland in Part II as a foil. The knights offer the two women a concrete representation of their transgression from the existing power structure. They suggest not only the extent of the two women's deviance from the patriarchy, but a way to reassume an orthodox social role. PMID- 11482426 TI - A double epidemic: crystal methamphetamine drug use in relation to HIV transmission among gay men. AB - Emerging research on methamphetamine use among gay men suggests that growth in the use of this drug could present serious problems for HIV/AIDS prevention within the gay community. This article summarizes current studies on the extent, role, and context of methamphetamine use among gay men and its relationship to high risk sexual behaviors related to HIV transmission. Methamphetamine is often used by gay men to initiate, enhance, and prolong sexual encounters. Use of the drug is, therefore, associated with particular environments where sexual contact among gay men is promoted, such as sex clubs and large "circuit" parties. Research with gay and bisexual men indicates that methamphetamine use is strongly associated with risky sexual behaviors that may transmit HIV. This relationship, coupled with emerging evidence that methamphetamine use is on the rise among gay men, suggests that the drug could exacerbate the HIV/AIDS epidemic among this community. The article offers recommendations for further research and suggestions for prevention programs regarding methamphetamine use by gay men. PMID- 11482427 TI - The Lesbian Internalized Homophobia Scale: a rational/theoretical approach. AB - This article reports the development and psychometric properties of a new scale that measures internalized homophobia in lesbians: the Lesbian Internalized Homophobia Scale (LIHS). This 52-item measure was developed using a rational/theoretical approach of test construction and includes five subscales. Research findings, based on a sample of 303 female participants, supported the reliability and validity of the LIHS in assessing internalized homophobia in lesbians. Implications for research and practice are discussed. PMID- 11482428 TI - The development of an Internalized Homonegativity Inventory for gay men. AB - Because existing scales measuring internalized homonegativity suffer from various content validity flaws, a more valid instrument is needed. The purpose of this study was to develop and provide preliminary reliability and validity evidence for the Internalized Homonegativity Inventory (IHNI), which measures internalized homonegativity in gay men. Forty-two items were initially generated from a construct definition and were administered to 241 gay men living in the U.S. Exploratory factor analyses suggested that 23 items loaded on 3 subscales, which were labeled Personal Homonegativity, Gay Affirmation, and Morality of Homosexuality. IHNI scores correlated in expected directions and magnitude with measures of extroversion, emotional stability, stage of gay identity development, and another internalized homonegativity instrument. Implications of the three factor structure of the IHNI for the conceptualization of internalized homonegativity are discussed. PMID- 11482429 TI - Ranking of stigmatization toward lesbians and their children and the influence of perceptions of controllability of homosexuality. AB - Recent custody rulings in this country have indicated that lesbian mothers are viewed more negatively than other parents with stigmatizing attributes, e.g., a history of mental illness or criminality. The current study compared willingness to engage in relationships with a variety of adults with potentially stigmatizing conditions (including lesbians) as well as the children of these stigmatized individuals among a sample of college students. The hypothesis that perceived controllability of homosexuality would be related to stigmatization of lesbians and their children was also tested. Findings included the following: (1) Participants reported a greater willingness to engage in relationships with children of stigmatized parents than with individuals who personally possess a stigmatizing attribute; (2) Participants reported a greater willingness to engage in relationships with adults with physically based stigmas and their children than with adults with mental/behavioral stigmas and their children, and (3) Participants who reported a belief that homosexuality is a controllable condition were less willing to engage in relationships with lesbians than respondents who reported a belief that homosexuality is uncontrollable. Discussion includes not only an evaluation of the results and limitations of the study, but also comments concerning judicial and societal stigmatization of lesbians and their children. PMID- 11482430 TI - Gay men with AIDS: perceptions of social support and adaptational outcome. AB - Using a multivariate design, this study examined social support as a mediating variable between the stressor of AIDS and adaptational outcome in a culturally and socioeconomically diverse sample of 60 urban gay men with AIDS. Whereas earlier studies have suggested that social support for gay men with AIDS is often lacking or ineffective, findings in this study indicated that family of origin and friends may provide more support to gay men with AIDS than has been generally recognized. Both family of origin and friend support were strongly correlated with positive outcomes, indicating that social support was not only forthcoming but was also effective. The relationship between family of origin support and positive outcomes was particularly strong. PMID- 11482431 TI - Dynamic simulation of electromechanical systems: from Maxwell's theory to common rail diesel injection. AB - This paper describes the state-of-the-art of dynamic simulation of electromechanical systems. Electromechanical systems can be split into electromagnetic and mechanical subsystems, which are described by Maxwell's equations and by Newton's law, respectively. Since such systems contain moving parts, the concepts of Lorentz and Galilean relativity are briefly addressed. The laws of physics are formulated in terms of (partial) differential equations. Numerical methods ultimately aim at linear systems of equations, which can be solved efficiently on digital computers. The various discretization methods for performing this task are discussed. Special emphasis is placed on domain decomposition as a framework for the coupling of different numerical methods such as the finite element method and the boundary element method. The paper concludes with descriptions of some applications of industrial relevance: a high performance injection valve and an electromechanical relay. PMID- 11482432 TI - The evolutionary position of turtles revised. AB - Consensus on the evolutionary position of turtles within the amniote phylogeny has eluded evolutionary biologists for more than a century. This phylogenetic problem has remained unsolved partly because turtles have such a unique morphology that only few characters can be used to link them with any other group of amniotes. Among the many alternative hypotheses that have been postulated to explain the origin and phylogenetic relationships of turtles, a general agreement among paleontologists emerged in favoring the placement of turtles as the only living survivors of the anapsid reptiles (those that lack temporal fenestrae in the skull). However, recent morphological and molecular studies have radically changed our view of amniote phylogenetic relationships, and evidence is accumulating that supports the diapsid affinities of turtles. Molecular studies favor archosaurs (crocodiles and birds) as the living sister group of turtles, whereas morphological studies support lepidosaurs (tuatara, lizards, and snakes) as the closest living relatives of turtles. Accepting these hypotheses implies that turtles cannot be viewed any longer as primitive reptiles, and that they might have lost the temporal holes in the skull secondarily rather than never having had them. PMID- 11482433 TI - How do general anaesthetics work? AB - Almost a century ago, Meyer and Overton discovered a linear relationship between the potency of anaesthetic agents to induce general anaesthesia and their ability to accumulate in olive oil. Similar correlations between anaesthetic potency and lipid solubility were later reported from investigations on various experimental model systems. However, exceptions to the Meyer-Overton correlation exist in all these systems, indicating that lipid solubility is an important, but not the sole determinant of anaesthetic action. In the mammalian central nervous system, most general anaesthetics act at multiple molecular sites. It seems likely that not all of these effects are involved in anaesthesia. GABAA- and NMDA-receptor/ion channels have already been identified as relevant targets. However, further mechanisms, such as a blockade of Na+ channels and an activation of K+ channels, also come into play. A comparison of different anaesthetics seems to show that each compound has its own spectrum of molecular actions and thus shows specific, fingerprint-like effects on different levels of neuronal activity. This may explain why there is no known compound that specifically antagonises general anaesthesia. General anaesthesia is a multidimensional phenomenon. Unconsciousness, amnesia, analgesia, loss of sensory processing and the depression of spinal motor reflexes are important components. It was not realised until very recently that different molecular mechanisms might underlie these different components. These findings challenge traditional views, such as the assumption that one anaesthetic can be freely replaced by another. PMID- 11482434 TI - Social encapsulation of beetle parasites by Cape honeybee colonies (Apis mellifera capensis Esch.). AB - Worker honeybees (Apis mellifera capensis) encapsulate the small hive beetle (Aethina tumida), a nest parasite, in propolis (tree resin collected by the bees). The encapsulation process lasts 1-4 days and the bees have a sophisticated guarding strategy for limiting the escape of beetles during encapsulation. Some encapsulated beetles died (4.9%) and a few escaped (1.6%). Encapsulation has probably evolved because the small hive beetle cannot easily be killed by the bees due to its hard exoskeleton and defensive behaviour. PMID- 11482435 TI - Cryptic mammalian species: a new species of whiskered bat (Myotis alcathoe n. sp.) in Europe. AB - The analysis of morphological, behavioural and genetic characters of whiskered bats revealed a new European bat species within the family Vespertilionidae. We describe the morphology, karyology, genetic similarity, ecology and distribution of Myotis alcathoe n. sp. It closely resembles Myotis mystacinus, Myotis brandtii and Myotis ikonnikovi in morphology, but all four species show clear genetic differences in two mitochondrial genes (ND1 and 12S rRNA). Myotis alcathoe n. sp. is the smallest species among the European whiskered bats and uses the highest frequency echolocation calls of all the European Myotis species. It prefers to hunt in small valleys with deciduous trees and flowing water, which is an endangered habitat. Records from Greece and Hungary indicate a distribution range in south-eastern Europe. PMID- 11482436 TI - Evaluation of the use of Sr2+ in alginate immobilization of cells. AB - In recent years the method of immobilization of living cells in Ca-alginate beads has gained a wide range of applications. In all cases high chemical stability of the immobilization material and mild conditions for the cells are prerequisites. However, in long-term experiments that may last for several days Ca-alginate may dissolve due to an exchange of Ca2+ with Na+, forming fluid Na-alginate. As well as Ca-alginate, the more chemically stable Sr-alginate and Ba-alginate are materials that have been used for the immobilization of living cells. In this study, the effects of Ca2+, Sr2+ and Ba2+ on growth, viability and intracellular free calcium concentration in a human leukemic T cell line (Jurkat) were investigated. The findings in this study, and the fact that Sr-alginate has a considerably higher chemical stability than Ca-alginate, led to the conclusion that Sr-alginate is a more suitable material for use in the entrapment of living cells in long-term studies. PMID- 11482437 TI - Cell-to-cell cross talk in the testis. PMID- 11482438 TI - Vaccines in urologic malignancies. AB - The prospect of activating the immune system to combat neoplastic disease has stimulated the interest of clinicians and scientists for over 100 years. Despite a few notable exceptions (especially with urologic malignancies), immunotherapy has not fully reached its considerable therapeutic potential for the treatment of cancer. Tumors undoubtedly express antigens that may act as targets for antitumor immunity, and advances in molecular biology and tumor immunology have recently revived the possibility of a cancer vaccine. This improved understanding has resulted in numerous successes with active immunotherapy in animal models and has facilitated the clinical testing of cancer vaccines. Ongoing advances in the identification of unique, tumor-specific antigens and their presentation to stimulate T cells will be necessary for the emergence of these novel vaccine therapies for cancer patients. Herein we review the current concepts of tumor immunology, including observations on cell types probably involved with the immune surveillance of tumors, the presentation and recognition of "foreign" antigens, and possible mechanisms of tumor escape from the immune response, all of which are critical to the understanding of new initiatives for cancer vaccine therapy. Finally, we review some of the successes and limitations of vaccine therapy for urologic malignancies. PMID- 11482439 TI - Early development of the gubernaculum and cremaster sac in estrogen receptor knockout mice. AB - AIM exogenous estrogen causes gubernacular atrophy and cryptorchidism in fetal rodents. Mice with an estrogen receptor-alpha (ERalpha) disrupted gene mutation (alphaERKO) were studied to determine whether ablation of endogenous estrogen action, through ERalpha, had an effect on gubernacular development. Serial sagittal sections were made of the pelvis in fetal and day 7 postnatal wild-type and alphaERKO mice with the estrogen receptor-alpha "knockout" gene mutation. Wild-type (n = 24), heterozygote (n = 13) and alphaERKO mice (n = 12) were sacrificed at 16, 17 and 18 days fetal life and at 7 days postnatally. The size of the gubernaculum, cremaster muscle, cremaster sac, and the width of the sac at both ends in day 7 mice were quantitated by computer analysis. Visually and statistically the ERKO mice could not be separated from the wild-type mice during fetal life. At day 7 postnatally, a thicker cremaster sac was noted morphologically, and also a statistically significant difference was seen in the width of the cremaster sac at the sac's tip. Sac area, cremaster muscle area and the width of the sac at the sac's end did not differ significantly. Overall there is minimal phenotypic change observed in the alphaERKO mouse compared to wild type at the early developmental stages investigated. However, at postnatal day 7, there is a difference in the width of the cremasteric sac tip. This suggests that the effect of ERalpha, and thus signaling on the developing gubernaculum, occurs late in development. Alternatively, an action from the recently discovered ERbeta may be involved. Exploration of a betaERKO and the double knock-out alphaERKO/betaERKO mouse should be informative in evaluating the effect of endogenous estrogens in gubernacular development. PMID- 11482440 TI - Immunocytochemical distribution of nitric oxide synthase in the human corpus cavernosum: an electron microscopical study using the tyramide signal amplification technique. AB - Nitric oxide has proven to be an important mediator in the relaxation of human cavernosal smooth muscle. Nevertheless, there are many inconsistencies in the literature regarding the cellular and subcellular distribution of endothelial nitric oxide synthase in the human penis. The purpose of this study was to reexamine the localization of eNOS and nNOS in the cellular anatomy of the human cavernous body by means of electron microscopical immunocytochemistry in combination with the tyramide signal amplification technique (TSA). Using specific antibodies against eNOS and nNOS, the NAPDH-diaphorase reaction and advanced protocols for fixation and staining procederes, the occurrence of NOS isoenzymes eNOS and nNOS were examined in cavernosal specimens of ten male patients who were subjected to surgery for penile deviation. eNOS immunoreactivity and NADPH-d staining was seen to be significantly present in the endothelial cells covering the cavernous spaces and in the endothelium of helicine arteries. In endothelial cells, the NADPH-d reaction product BSPT formazan was abundantly detectable attached to membranes of the endoplasmatic reticulum and the mitochondria whereas posititve eNOS immunostaining was seen in the endothelial cells throughout their cytoplasm without any particular relation to organelles. No considerable eNOS immunoreactivity was detectable in the trabecular smooth muscle cells. nNOS staining was found in nerve fibers innervating the cavernous body and cavernosal arteries. Our results counteract the hypothesis of the cavernous smooth muscle as a local source of NO and underline the importance of an intact endothelial function for penile erection and the contribution of eNOS to this process. PMID- 11482441 TI - Testosterone reaction after testicular biopsies--further investigation in the normogonad and cryptorchid rat model. AB - From the follow-up of patients undergoing extensive testicular sperm extraction, the question of a consecutive decrease in testosterone levels arose. Further investigation was performed in the normogonad and cryptorchid Sprague-Dawley rat model. From groups A (n = 40 normogonad rats) and B (n = 40 cryptorchid rats), eight animals were taken and the following surgical interventions were performed: general anaesthesia (A-0, B-0), scrotal exploration (A-1, B-1), one testicular biopsy with a 2-mm tunica incision (A-2, B-2), two biopsies with a 4-mm incision (A-3, B-3), and four biopsies with a 2-mm incision (A-4, B-4). Standardised testosterone control was performed 1 day before as well as 1, 7 and 14 days after surgery. No specific testosterone reaction was found in A-0, an insignificant decrease in A-1 (3.8 nmol/l), a significant decrease in A-2 (6.27 nmol/l), in A-3 (12.96 nmol/l) and in A-4 (11.88 nmol/l). In the B rats, even in B-3 and B-4 no significant decrease was measured. The testosterone reaction correlated to the amount of tissue that was extracted, but not to the number of biopsies. The cryptorchid rats did not react in a more sensitive manner. PMID- 11482442 TI - A new experimental model for cryptorchidism: inguinoscrotal approach. AB - We investigated the effect of inguinal canal closure as a new mechanically induced cryptorchid rat model. The effectiveness of this new model was evaluated by histopathological examination. Thirty-one 21-day-old Wistar rats were divided into four groups. In groups 1 (n = 6), 2 (n = 6) and 3 (n = 7), unilateral undescended testis was created by performing inguinal canal closure with inguinoscrotal approach. Sham-operated rats were used as controls in group 4 (n = 12). The rats were killed on day 30 after surgery in group 1, day 45 in group 2 and day 60 in group 3. The seminiferous tubular diameter, number of tubules with mature germ cell and Leydig cell clusters were evaluated. None of the rats were lost during the study period. Signs of infection were not detected in operation site although antibiotics were not used. Overall only three (16%) testes descended into scrotum in study groups. The operation time was 3-4 min for each rat. Histopathological examination revealed detrimental effects of cryptorchidism on testicular growth in study groups. In all groups, except the sham group, the mean tubular diameter and the number of tubules with mature germ cells in the left testicle were significantly decreased compared to the right ones. Our findings were in correlation with other experimental studies using different rat models of cryptorchidism. This new model of cryptorchidism is considered to provide a simple and effective technique for investigating the impaired development of the testes in cryptorchidism. PMID- 11482443 TI - Relaxant effects of some benzothiazolinone derivatives on isolated rabbit corpus cavernosum. AB - In the present study, two 6-(fluorobenzoyl)-3-piperazinomethyl-2 benzothiazolinone derivatives were synthesized and their relaxant effects on isolated rabbit corpus cavernosum investigated. Compounds Y-16 and Y-21 can alter the ability of corpus cavernosum smooth muscle to contract. Strips of rabbit corpus cavernosum smooth muscle were mounted in isolated tissue baths for measurement of isometric contractile force. Compounds (10(-6) - 10(-3) M) did not cause contraction but induced relaxation in precontracted corpus cavernosum smooth muscle. Neither N-nitro-L-arginine methylester (L-NAME) nor indomethacin affected the relaxant effect of these compounds. Glibenclamide and tetraethylammonium chloride (TEA) also did not influence the relaxation induced by the compounds. In conclusion, in isolated rabbit corpus cavenosum, Y16 and Y21 have a relaxant potency equal or superior to known vasoactive agents. Further investigations are needed to show the importance of these effects for the diagnosis and treatment of erectile dysfunction. PMID- 11482444 TI - Carnitine as a preventive agent in experimental renal ischemia-reperfusion injury. AB - Reactive oxygen species generated during the reperfusion of ischemic kidney, as well as any other tissue, cause lipid peroxidation damaging the cell membrane. The aim of this study was to investigate the effect of carnitine in reperfusion injury of the kidney. Male albino rabbits were subjected to unilateral renal 1-h warm ischemia followed by 15 min of reperfusion. Group I (n = 9): control group received 3 cc of isotonic saline solution and group II (n = 9): carnitine group received 100 mg/kg of carnitine. Blood samples were collected at the 15th min of reperfusion from the left renal vein selectively. Preischemic and post reperfusion serum and renal tissue MDA levels were measured by thiobarbituric acid reactive substances (TBARS) spectrophotometric analysis. The preischemic serum and tissue MDA values (sham values) for groups I and II were statistically comparable (P > 0.01). Serum and tissue MDA levels were markedly elevated after 15 min of reperfusion in group I (P < 0.01), while the values remained in the baseline levels following reperfusion in group II (P > 0.01). In group I, the major histological differences observed in the reperfused kidneys were marked edema and congestion whereas glomerular and tubular cellular integrity were well preserved in group II. Pre-treatment with carnitine in solid organ transplantations, preschock states, surgical procedures that require temporary vascular clamping etc. may be helpful to minimize the reperfusion injury in the involved tissue, reducing morbidity and mortality. PMID- 11482445 TI - The effect of caffeic acid phenethyl ester on ischemia-reperfusion injury in comparison with alpha-tocopherol in rat kidneys. AB - Oxygen-derived free radicals have been implicated in the pathogenesis of renal injury after ischemia-reperfusion. Caffeic acid phenethyl ester (CAPE), an active component of propolis extract, exhibits antioxidant and anti-inflammatory properties. To determine whether CAPE offers any advantage over alpha-tocopherol, we compared their effects on an in vivo model of renal ischemia-reperfusion injury in rats. CAPE at 10 micromol/kg or alpha-tocopherol at 10 mg/kg was administered intraperitoneally before reperfusion. Acute administration of CAPE suppressed ischemia-reperfusion induced renal lipid peroxidation and tissue injury more than alpha-tocopherol. CAPE may therefore offer a therapeutic advantage in acute injury settings. PMID- 11482446 TI - Increased calcium oxalate crystal nucleation and aggregation by peroxidized protein of human kidney stone matrix and renal cells. AB - Kidney stone matrix protein fractions eluted from DEAE cellulose column showed increased oxalate binding activity and had negative correlation with reduced thiol content. Fraction I (eluted in Tris-HCl, pH 7.4) and fraction 3 (0.3 M NaCl in buffer) showed nucleation and aggregation-promoting properties while fraction 2 (0.05 M NaCl in buffer) showed an inhibitory effect in an in vitro crystallization system. On peroxidation, fractions 1 and 3 showed a further increase in the promoting effect whereas fraction 2 showed a reduction in the inhibitory effect of nucleation and aggregation of calcium oxalate crystals. Protein peroxidation was negatively correlated with the inhibitory activity of the protein on calcium oxalate nucleation and aggregation. A similar promoting effect of nucleation and aggregation was seen with mitochondria and nucleus after peroxidation. These studies suggested that peroxidation of protein or tissue had greater influence on the nucleation and aggregation property of calcium oxalate crystal growth. PMID- 11482447 TI - Tobacco use and occupational exposure to carcinogens, but not N-acetyltransferase 2 genotypes are major risk factors for bladder cancer in the Japanese. AB - Our study investigated the risks of genotypes of N-acetyltransferase 2 (NAT2), tobacco use and/or occupational exposure to carcinogens in patients with bladder cancer and in age- and sex-matched controls in Japanese. NAT2 genotypes were categorized into two groups, homozygous mutant (slow acetylator genotype) and homozygous and heterozygous wild type (fast acetylator genotype). The percentage of NAT2 slow acetylator types was 6.7% in the bladder cancer patients, close to the value for controls (6.1%). There was no association between NAT2 slow acetylator genotype and the risk of bladder cancer. This association was also insignificant when subjects were restricted to those who used tobacco or those occupationally exposed to carcinogens. In contrast, tobacco use in combination with exposure to carcinogens was a significant risk factor, as based on the odds ratio and chi-square test. The combination of both factors should be an additive risk factor for bladder cancer. In this study, we demonstrated that the environmental factors of smoking habit and occupational exposure for carcinogenicity are much more important than genetic factors in bladder cancer. PMID- 11482448 TI - Effect of photodynamic therapy on urinary bladder function: an experimental study with rats. AB - Photodynamic therapy (PDT) produces localized necrosis with light after prior administration of a photosensitizing drug. The problems with laser light dosimetry and complications relating to bladder function appear to be important limiting factors of PDT in urology. Photodynamic therapy on urinary bladder with normal epithelium of rats was performed using an argon ion laser as an energy source, with aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) photosensitizer. Four hours after ALA intravenous administration, the bladders were intravesically radiated with light doses 20, 40, or 80 J/cm2. Animals in the control group did not receive ALA and were radiated with 20 J/cm2 light dose. Three weeks prior to PDT, the bladder capacity and pressure changes during filling cystometry were assessed. Cystometrics were repeated 1, 3, 7, or 21 days after laser therapy. The light dose 20 J/cm2 and 40 J/cm2 together with the used ALA dose caused no reduction in bladder capacity, whereas 80 J/cm2 light dose produced up to 50% reduction in the capacity at 3 weeks postoperatively. In control group without ALA, the animals did not regain more than 34% of the capacity of their control values at 3 weeks. The light dose of 20 J/cm2 and 40 J/cm2 with ALA induced functional changes that subsided after day 1. Our results indicate that with proper dosing of photosensitizing drug and light energy, the functional impairment of urinary bladder may be reduced as transient. These findings support the use of PDT as safe therapy of superficial bladder cancer. PMID- 11482449 TI - The effects of organ resection on rat urinary bladder carcinogenesis. AB - The effect of unilateral nephrectomy, orchiectomy or partial hepatectomy on the growth of chemically induced rat bladder tumors was investigated. Male F344 rats were treated with 0.05% N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) for 5 weeks, and surgical resection of one of these organs was performed 2 weeks after the completion of BBN administration. Histological evaluation of the bladder 24 weeks after the start of the experiment revealed that unilateral nephrectomy and orchiectomy significantly increased the numbers of preneoplastic and neoplastic lesions as compared with the corresponding sham-operated groups. Partial hepatectomy also enhanced tumor growth, although not significantly. Immunohistochemical studies examining the effect of organ resection on normal bladder urothelium showed that BrdU immunostaining of urothelial cells significantly increased 7 days after unilateral nephrectomy or orchiectomy, while BrdU incorporation was minimum after partial hepatectomy or sham operation. C-met expression in the bladder urothelium was evident following unilateral nephrectomy or partial hepatectomy, while increased immunoreactivity of androgen receptor was noted following unilateral orchiectomy. Further study is needed to determine the exact mechanism of the bladder tumor growth-enhancing effect associated with organ restriction. PMID- 11482450 TI - Radioimmunoscintigraphy using an anti-prostate monoclonal antibody (E4): a dosimetric evaluation. AB - The aim of this study was to evaluate different strategies to increase the tumour radiation dose for experimental radioimmunotherapy using 125I-labelled monoclonal antibody (MAb) E4 in a nude mice model xenografted with DU-145 tumours. The effects from a single injection of the 125I-labelled MAb E4, the same total amount of radiolabelled MAb E4 divided into three repeated injections, and the effect of pre-targeting with non-labelled MAb E4 for reducing the amount of shed antigen were investigated. Based on repetitive quantitative radioimmunoscintigraphies, calculation of the tumour radiation dose delivered from the 125I-nuclide was performed for each strategy. The single injection strategy without pretargeting rendered the highest mean tumour radiation dose, i.e. 0.23 Gy/MBq. Pretargeting with non-labelled MAb E4 before a single injection of [125I]E4 resulted in a slightly lower mean tumour radiation dose, i.e. 0.19 Gy/MBq, compared to the single injection alone. An even lower mean tumour radiation dose, i.e. 0.14 Gy/MBq, was obtained when the same total administered amount of activity was divided into three separate injections given in 10-day intervals. We concluded that the single injection strategy is the most efficient when using MAb E4 in this tumour model. The tumour radiation doses were not increased by dividing the same amount of activity into three injections or by pretargeting with non-labelled MAb E4. PMID- 11482451 TI - Chemosensitivity of prostatic tumour cell lines under conditions of G2 block abrogation. AB - Conventional chemotherapy has had very limited success in the control of hormone refractory prostate cancer. Methylxanthine derivatives, such as pentoxifylline (PTX), are known to abrogate the G2 block and enhance the toxicity of ionising irradiation and chemotherapeutic agents. It is now also established that late addition of the cytotoxic drug after irradiation under conditions of G2 block abrogation sensitises human tumour cells for cytotoxins. Here we assess whether the chemosensitivity of prostate tumour cell lines can be enhanced by the application of a low dose of drug in conjunction with a G2 block abrogator. Prostate cell lines DU145, BM1604 and LNCaP were irradiated with 7 Gy 60Co gamma irradiation. A sub-toxic (2 mM) dose of pentoxifylline and a cytotoxic drug were added at maximum expression of the G2 cell cycle block and cell survival was determined by colony assay. Cisplatin, etoposide and vinblastine were tested at a toxic dose of 10% (TD10). In the TP53 mutant cell lines, DU145 and BM1604, dose enhancement factors (EFs) were found to be in the region of 4.20 for cisplatin, 3.70 for vinblastine, and 3.20 for etoposide. In the TP53 wild-type cell line, LNCaP, the enhancement factors were low and in the region of 1.20 for cisplatin, vinblastine and etoposide. It is clear, therefore, that toxicity enhancement factors (EFs) are greater in the TP53 mutant cell lines, DU145 and BM1604, than in the TP53 wild-type cell line, LNCaP. The results indicate that a significant enhancement of drug toxicity can be obtained if the cytotoxic drug is given under conditions of G2 block abrogation. The sensitisation of prostate cancer cells to cytotoxic drugs is particularly high in radiation-resistant TP53 mutant tumour cells. Drugs which abrogate G2 block have the potential to enhance the therapeutic index and therefore reduce the toxicity of chemotherapy drugs. PMID- 11482452 TI - Secretion of TNF-alpha, IL-8 and nitric oxide by macrophages activated with Agaricus blazei Murill fractions in vitro. AB - Water extracts of the mycelial culture and fruiting bodies of Agaricus blazei Murill were fractionated by ethanol precipitation using various ethanol concentrations. Original water extracts from mycelia (Fraction A-0) and fruiting bodies (Fraction B-0) induced TNF-alpha secretion by macrophages derived from rat bone marrow. Fractions B-4 and B-5 obtained from ethanol precipitation of fruiting bodies using 44% and 50% ethanol, respectively, and Fraction B-6 obtained from the supernatant at 50% ethanol markedly induced TNF-alpha secretion. Similar effects were observed in IL-8 secretion by macrophages. Regarding nitric oxide (NO), Fraction B-5 induced a significant increase in NO secretion and Fractions B-4 and B-6 induced slightly NO secretion. Northern blot analysis showed that the increases in cytokine- and NO secretion were due to an increase in cytokine mRNAs or NO synthase mRNA. Therefore, it is concluded that Agaricus blazei Murill components which activate macrophages result in the induction of cytokine- and NO secretion in vitro. PMID- 11482453 TI - Occludin regulates actin cytoskeleton in endothelial cells. AB - Occludin is a major membrane component of tight junctions of endothelial cells, though the role of this molecule is not fully understood. RLE cells, derived from rat lung endothelial cells, express a negligible level of occludin with clear expression of E-cadherin and ZO-1 at cell junctions. Introduction of occludin by transfection induced clear junctional expression of occludin with few or no changes of expression of E-cadherin and ZO-1. The paracellular barrier function, as determined by transelectrical resistance and flux of non-ionic small molecules, was not detectably upregulated. When cells expressing occludin were cocultured with RLE cells null for occludin, clear junctional expression of occludin was observed irrespective of the expression of occludin on the apposing cells. Cortical actin was developed at the site of these occludin positive cell junctions. Treatment of cells with an actin depolymerizing agent, mycalolide B, abolished junctional expression of occludin together with E-cadherin and circumferential actin. ZO-1 showed relative resistance to this actin depolymerizing treatment and was maintained at the cell junctions, though fragmentation of immunoreactivity was detectable. Collectively, junctional expression of occludin was not associated with paracellular barrier function in this cell line. There was, however, a close correlation of occludin with the actin cytoskeleton, indicating a role of occludin as an important molecule in the regulation of the actin cytoskeleton in endothelial cells. PMID- 11482454 TI - Localization of plectin and other related proteins along the sarcolemma in smooth muscle cells of rat colon. AB - Plectin is a versatile linker protein which is associated with various types of cytoskeletal components and/or filaments including intermediate filaments. To better understand the functional roles of plectin in smooth muscle cells, we examined the distribution of plectin and other related proteins in rat colon smooth muscles by confocal laser and electron microscopy. The sarcolemma of smooth muscle cells exhibits two ultrastructurally distinct domains, domains associated with dense plaques and caveola-rich domains. Staining with anti plectin and anti-desmin antibodies showed that plectin was localized along the sarcolemma in an intermittent manner and desmin was distributed in the sarcoplasm and intermittently at the cell periphery where it was codistributed with desmin. Plectin exhibited complementary and non-overlapping distribution to caveolin-1 and dystrophin, components of caveola domains, whereas plectin was codistributed with vinculin, talin and integrin beta1, components of dense plaques. Plectin was also codistributed with beta2-chain laminin but not with beta1-chain laminin. Electron microscopic observations on the sarcolemma revealed close association of intermediate filaments with dense plaques. Correlated confocal and electron microscopy clearly demonstrated that anti-plectin fluorescence corresponded to dense plaques but not to caveola domains in electron microscopic images. These findings indicate that plectin is confined to dense plaques to which desmin intermediate filaments may be anchored in rat colon smooth muscle cells. PMID- 11482455 TI - Nuclear deviation in hepatic parenchymal cells on sinusoidal surfaces in Arctic animals. AB - In normal rat and human, most of the nuclei of hepatic parenchymal cells are centrally located in the cytoplasm. However, it is reported that the nuclei of hepatic parenchymal cells are situated at a deviated position on sinusoidal surfaces under pathological situations such as chronic hepatitis, hepatocellular carcinoma, adenomatous hyperplasia, or regeneration. During a study on the mechanism of extreme vitamin A-accumulation in hepatic stellate cells of arctic animals including polar bears, arctic foxes, bearded seals, and glaucous gulls, we noticed that these arctic animals displayed the nuclear deviation in hepatic parenchymal cells on sinusoidal surfaces. In this study, we assessed the frequency of hepatic parenchymal cells showing the nuclear deviation on the sinusoidal surfaces in arctic animals. A significantly higher frequency of the nuclear deviation in hepatic parenchymal cells was seen in polar bears (89.8+/ 3.4%), arctic foxes (68.6+/-10.5%), bearded seals (63.6+/-8.4%), and glaucous gulls (24.2+/-5.8%), as compared to that of control rat liver (9.8+/-3.5%). However, no pathological abnormality such as fibrosis or necrosis was observed in hepatic parenchymal and nonparenchymal cells of arctic animals, and there were no differences in the intralobular distribution of parenchymal cells displaying the nuclear deviation in the livers from either arctic animals and control rats. The hepatic sinusoidal littoral cells such as stellate cells or extracellular matrix components in the perisinusoidal spaces may influence the nuclear positioning and hence the polarity and intrinsic physiological function of parenchymal cells. PMID- 11482456 TI - Proliferation stage-dependent expression of DNA methyltransferase (Dnmt1) in mouse small intestine. AB - In cultured cells, the maintenance-type DNA methyltransferase (Dnmt1) is highly expressed during the proliferation stage. In the present study, we detected significant expression of Dnmt1 protein in the nuclear fraction of mouse small intestine. From its mobility in SDS polyacrylamide gel electrophoresis and the specific antibodies against the somatic cell-type Dnmt1, Dnmt1 was determined as a somatic cell type. Immunofluorescence study revealed that the Dnmt1 was highly expressed in the proliferating stem cells in crypts, and was localized in the nuclei. The present results indicate that the expression of Dnmt1 in vivo is also under the control of cell proliferation as in cultured cells. PMID- 11482457 TI - Ultrastructural modifications and phosphatidylinositol-3-kinase expression and activity in myocardial tissue deriving from rats in different experimental conditions. AB - Oxygen supply is essential in the maintenance of the physiological cell metabolism. In fact, both lower and higher O2 concentrations induce modifications of the enzymatic activity of the cell which determine, in turn, morphological changes at nuclear and cytoplasmic level. Among the molecules involved in the maintenance of the cellular homeostasis, the signal transduction pathway PI-3 kinase/AKT-1 should be included. Here we suggest a relationship between the modulation of this pathway and the morphological modifications occurring "in vivo" in myocardial tissue upon hypoxic and hyperoxic stress. In particular, down regulation of this pathway, which when activated is known to deliver an anti apoptotic signal, is concomitant to the maintenance of the apoptotic events occurring in these cells in response to oxidative stresses. PMID- 11482458 TI - Stromal cells provide signals different from cytokines for STAT5 activation in hematopoietic cells. AB - After detachment from the stromal cells, hematopoietic stem cells are thought to differentiate to the cytokine-dependent stages where their growth and differentiation are promoted by these cytokines. To examine the stromal regulation of hematopoietic stem cells, we previously established a primitive hematopoietic stem-like cell line, THS119, whose growth was dependent on the bone marrow stromal cell line, TBR59, and from which IL-3- (THS119/IL-3) or IL-7- (THS119/IL-7) dependent cell lines were then generated. Using these cell lines, we examined the difference in signals mediated by the stromal cells and cytokines. The cytokine-dependent cell lines (THS119/IL-3 and THS119/IL-7) showed induction of STAT5 phosphorylation and target genes for STAT5 such as CIS, pim-1, p21 and bcl-xL upon addition of IL-3 or IL-7. IL-3 or IL-7 also induced STAT5 phosphorylation and STAT5 target genes of the stromal cell-dependent cell line, THS119, in the absence of stromal cells at levels similar to the cytokine dependent cell lines. However, quite interestingly, TBR59 stromal cells could not induce STAT5 phosphorylation of THS119 cells, although they did induce STAT5 target genes in THS119 cells. In addition, the mRNAs for STAT5 target genes in THS119 cells on the stromal cells seemed to be more stable than those in the cytokine-dependent cell lines. Expression of the antiapoptotic genes bcl-2 and bcl-xL was higher in the stromal cell-dependent cell line than in the cytokine dependent cell lines. These results suggested that stromal cells and cytokines may provide different signals for growth and differentiation of the hematopoietic cells. PMID- 11482459 TI - Knee and ankle: human joints with different susceptibility to osteoarthritis reveal different cartilage cellularity and matrix synthesis in vitro. AB - Clinical experience shows that symptoms and pathological changes of primary osteoarthritis (OA) are more frequent and severer in the knee than in the ankle joint. The different anatomy of both weight-bearing joints implies that biomechanical differences may contribute to their varying susceptibility to OA. This study aims at elucidating other non-biomechanical factors to explain these fundamental differences in secondary OA prevalence. Human cartilage of matched ankle and knee joints from organ donors was dissected in full-thickness slices or in layers. The DNA content for estimation of cell number was analyzed fluorometrically. Chondrocytes were cultured in organ culture or after isolation in alginate. Proteoglycan synthesis was determined by 35S incorporation, and collagen synthesis by 3H-proline incorporation. This study demonstrates that in both joints, the cell density sharply declines between newborn and young infant ages. In addition, cartilage from the ankle joint is significantly more cellular than cartilage from the knee joint. In general, ankle chondrocytes synthesize more proteoglycans (PGs) and collagens than knee chondrocytes, and deep zone chondrocytes more than superficial zone chondrocytes. The biochemical properties of chondrocytes of the ankle and knee joints differ significantly and might play an important role in the pathogenesis of OA. PMID- 11482460 TI - Selective thigh muscle atrophy in trans-tibial amputees: an ultrasonographic study. AB - In trans-tibial amputees, PTB (patellar tendon bearing) prostheses provide almost physiological mobility of the knee joint in the sagittal plane. Nevertheless, there are characteristic adaptations of the knee joint muscles. Myosonography is a suitable method for depicting muscle atrophy and hypertrophy due to muscle dysfunction. The present study was intended to assess anatomical alterations of thigh muscles in trans-tibial amputees wearing a PTB prothesis. Thicknesses and cross-sectional areas of the quadriceps femoris, sartorius, gracilis, semitendinosus and biceps femoris muscles were determined ultrasonographically on both limbs in 17 amputees with a PTB prothesis. The gait was analysed using an optoelectronical system, force plates and surface electromyography of the vastus lateralis and biceps femoris muscles. Quadriceps femoris and sartorius muscles of the amputated extremity exhibited significant atrophy compared with the contralateral limb (reduction of muscle thickness ranged between 11.7% and 30.4%), whereas the gracilis and hamstring muscles were not significantly affected. Even the quadriceps femoris muscle of the non-amputated limb showed a slight atrophy compared with a reference group. Increased echointensities were found predominantly in the quadriceps muscle on the amputated leg. During gait, electromyographical activity within the amputated limb was reduced in the vastus lateralis and increased in the biceps femoris muscle. Even long-term adaptation to PTB prostheses results in characteristic deviation from normal gait. Atrophy occurs in the ventral thigh muscles, predominantly on the amputated leg, whereas the dorsal thigh muscles are hardly affected, probably due to compensatory hyperactivity. PMID- 11482461 TI - Scapular neck fracture--the influence of permanent malalignment of the glenoid neck on clinical outcome. AB - A scapular neck fracture is considered unstable if it is associated with an ipsilateral clavicular fracture or an acromioclavicular (AC) joint dislocation. Currently, it is recommended that stabilization of a disrupted shoulder girdle must be achieved through open reduction and internal fixation of the clavicular fracture or by reduction of the AC joint, without addressing the scapular neck. However, if the displaced glenoid neck is not simultaneously reduced, malalignment of the glenoid neck may persist. The purpose of this retrospective study was to analyze the effect of associated shoulder girdle injury on glenoid displacement and the influence of glenoid malalignment on clinical outcome. Nineteen patients with scapular neck fractures were reviewed clinically and radiologically at a mean of 8 years (range 2-21 years) after injury. None of them has developed nonunion of the scapular neck, and only one showed radiological signs of mild degenerative joint disease. The glenopolar angle (GPA), which assesses the rotational malalignment of the glenoid about an anteroposterior axis perpendicular to the scapular plane on plain X-rays was measured less than 20 degrees in six patients. Three of them had sustained an associated clavicular fracture or AC joint dislocation. The other 3 patients had permanent severe malalignment of the glenoid neck in the absence of an associated shoulder girdle injury. Five patients with GPA less than 20 degrees complained of moderate or severe pain, whereas of the 13 patients with mild or no glenoid rotational displacement or medial displacement alone, 11 patients had no or mild pain, and only 2 had moderate or severe pain (P = 0.0095). Five patients presented with reduced activities of daily living, 4 of them had severe glenoid rotational displacement (P = 0.0173). Loss of motion was found in only 2 patients, and both had a severely displaced glenoid neck (P = 0.088). In conclusion, severe displacement of the glenoid neck may occur with or without associated fracture of the clavicle or dislocation of the AC joint and can be identified as a GPA less than 20 degrees. Scapular neck fractures with such malalignment have a less favorable long-term outcome compared with otherwise comparable cases with absence of glenoid malalignment as measured with the glenopolar angle. PMID- 11482462 TI - Functional results after anterior cruciate ligament reconstruction using the patellar ligament bone-tendon-bone technique. AB - This study evaluates the outcome after surgical repair of the cruciate ligament in 129 patients (physical examination plus questionnaire) and another 49 patients (questionnaire only) after a mean interval of 3.2 years. A modified Lysholm score and the OAK score together with KT 1000 arthrometry were used. Ninety-five percent of patients evaluated the overall result as 'excellent' or 'good'. The overall OAK objective score was good or excellent in 81%. Elongation as measured by KT 1000 arthrometry was 1.0 mm (67 N) and 1.2 mm (90 N) on average compared with the contralateral knee. PMID- 11482463 TI - Mid-term results of the cementless CLS stem. A 7- to 11-year follow-up study. AB - Between 1986 and 1990, 335 primary total hip arthroplasty (THA) were performed using the cementless CLS stem. The mean age at surgery was 55.3 years. The retrospective study was performed at an average follow-up time of 8.9 years. A total of 232 patients (69.3%) underwent clinical and radiological examination, 33 patients (9.9%) were interviewed by telephone, and 9 stems (2.7%) had been replaced meanwhile. The mean Harris hip score was 91.7 points, and the mean Merle D'Aubigne score was 16.8 points. One stem had subsided during the first postoperative year. Ectopic ossifications classified as Brooker III were noted in 5.7%, with no Brooker IV being seen. Radiolucent lines were found in 43.5%, and 11% showed enlargement with time. Focal osteolyses were present in 36%, of which 24% progressed with time. At the latest follow-up, we did not find any stem loosening clinically or radiologically. PMID- 11482464 TI - The influence of fracture etiology and type on fracture healing: a review of 104 consecutive tibial shaft fractures. AB - This study was conducted to determine retrospectively the factors which influence fracture healing and risk for nonunion in patients with tibial shaft fractures. One hundred consecutive patients with 104 tibial shaft fractures and a mean age of 40 (14-85) years were reviewed. Fractures were classified according to the AO classification system. There were 22 open fractures and 52 comminuted fractures. Thirty-eight fractures were caused by high-energy trauma. Fracture pattern, soft tissue condition, level of energy of the trauma, malalignment, and treatment methods were identified. The influence of these factors on the time of hospital stay and sick-leave, delayed union, and nonunion were calculated. Normal healing occurred in 61 fractures with a mean healing time of 17 weeks, delayed union in 27 with a mean of 35 weeks, and nonunion in 16 with a mean of 69 weeks. The relative risk of developing nonunion in open fractures was 8.2 (CI = 2.9-10.5) and 2.9 (CI = 1.2-3.2) in fractures exposed to high-energy trauma. This study showed that the soft-tissue condition and level of trauma energy are good predictors for the development of complications. Considering these risk factors at an early stage in the planning of treatment might reduce the risk of nonunion. ergy level of the trauma, fracture comminution, initial fracture displacement, treatment method, contamination, and associated injuries will influence fracture healing. Conservative treatment has in general been recommended for undisplaced closed or grade I open fractures caused by low-energy trauma [15, 18, 21]. However, conservative treatment of a tibial shaft fracture means immobilization of the lower leg for a long period of time, especially if healing is delayed. Thus, it is important to consider the risk of healing complications when planning the appropriate treatment method in the early stage after the injury. The purpose of this study was to analyze complications such as delayed union and nonunion and to identify factors which affect the healing of tibial shaft fractures. PMID- 11482465 TI - Surgical treatment of resistant tennis elbow. A prospective, randomised study comparing decompression of the posterior interosseous nerve and lengthening of the tendon of the extensor carpi radialis brevis muscle. AB - We compared decompression of the posterior interosseous nerve (PIN) and lengthening of the distal tendon of the extensor carpi radialis brevis (ECRB) for treatment of tennis elbow in a randomised trial of 28 patients. Fourteen underwent decompression of PIN and 14, lengthening of ERCB. The groups did not differ significantly with regard to age, sex and work activities. The average duration of preoperative symptoms was 23 months. The PIN was exposed in the groove between the brachioradialis and brachialis muscles and decompressed at the arcade of Frohse by means of a 1-2 cm incision through the supinator muscle. The ECRB tendon was lengthened by Z-plasty at the dorsilateral aspect of the forearm. No postoperative complications occurred. The outcome after the primary operation was successful in 50% of the PIN group and in 43% of the ECRB group. Four of the 5 patients with a poor outcome were reoperated in the former group and 3 in the latter. The overall outcome after a mean follow-up of 31 months after the primary operation was successful in 60% of the cases. PMID- 11482466 TI - Deleterious effects of local corticosteroid injections on the Achilles tendon of rats. AB - The purpose of this study was to examine the pathological changes in the Achilles tendon and its paratenon after intratendinous corticosteroid injections and to reveal the effects of this drug on healthy tendon. We also sought for the effects of these injections compared with compression with a clamp on the Achilles tendons of the rats. Fifty-two Achilles tendons in 26 male Wistar rats were included in the study. Betamethasone injections were applied to the left tendons at different intervals, while the right tendons served for compression with mosquito clamps for varied periods. At the end of 30 days, all of the tendons were excised and examined histopathologically according to a semiquantitative scoring system. Histopathologic evaluation demonstrated some degree of degeneration in both groups. Statistical analysis showed no significant difference among the two groups, but in macroscopic evaluation, the tendons in the betamethasone group demonstrated enlargement and strong adhesion to the subcutaneous tissue. We conclude that intratendinous betamethasone injections are as harmful as compression with a clamp and can be used as a degeneration producing model in further studies. Enlargement of the tendon mass and strong adhesion to the subcutaneous tissue can be due to injection of the betamethasone partly outside the tendon. PMID- 11482467 TI - Radiological signs of osteitis around extramedullary metal implants. A radiographic-microbiological correlative analysis in rabbit tibiae after local inoculation of Staphylococcus aureus. AB - Radiographic changes in the early stages of osteomyelitis may be subtle and, especially after plate osteosynthesis, frequently missed. A previously described experimental model of local bacterial infection was used in an attempt to determine the reliability of specific changes on conventional radiographs for the diagnosis of osteitis after metal-plate implantation and subsequent inoculation of Staphylococcus aureus in rabbit tibiae. Roentgenograms of the treated limbs were evaluated, and seven radiographic parameters, to which numerical scores were assigned, were determined for each bone. Our results substantiate the conclusion that a radiographically verified periosteal reaction is a constant and early skeletal feature of acute osteomyelitis and has the strongest association to the microbiological results (P < 0.05), emphasising its high predictive value. Plate implantation does not notably impede the diagnosis of osteomyelitis. An association between the amount of inoculated bacteria and the extent of radiographic changes could be found. The results of this present study closely resemble those described in man and suggest that this model may be useful for future experimental investigations in determining a score judging the severity of osseous involvement in local bacterial infection after plate osteosynthesis. PMID- 11482468 TI - A technique for accurate reproduction of the femoral anteversion during primary total hip arthroplasty. AB - Accurate reproduction of the femoral anteversion is important in achieving stability of the prosthetic joint in total hip arthroplasty (THA), while placement of the femoral stem in excessive anteversion or retroversion results in an increased incidence of dislocation. Due to a number of factors, the current surgical technique results in significant variation between the anatomical and the surgically reconstructed femoral anteversion. This technical note describes a method to reproduce the normal anatomical femoral anteversion accurately in any case of primary THA. The method is based on the idea that the initial stages of femoral canal preparation can be completed before the femoral neck is osteotomised and therefore at a stage when the femoral head and neck are still accessible and can be used as landmarks for accurate reconstruction of the femoral anteversion. The described technique for accurate reconstruction of the anatomical version of the femoral head is indicated for all cases of primary THA where reproduction of the normal anatomical centre of rotation of the hip joint is a principal aim. PMID- 11482469 TI - Correlation between the Q angle and the patella position: a clinical and axial computed tomography evaluation. AB - The purpose of this study was to evaluate the significance of the Q angle with respect to the patella position. Fifty-six knee joints of 34 patients (15 bilateral) with chronic patellofemoral pain were prospectively evaluated. All patients were examined by the same orthopaedic surgeon (R.M.B.) and the Q angle measured clinically and using long radiographs. Additionally, axial computed tomography (CT) scans were obtained through the center of the patellar articular cartilage in 0 degrees of flexion. Three different patellofemoral indices were measured by the second author (K.W.), who was not involved in the clinical examination: lateral patellar displacement (LPD), lateral patellar tilt (LPT), and patella-lateral condyle index (PLCI). These results were compared with the values of the measured Q angle. For statistical analysis, the Pearson correlation coefficient was calculated and the Statistical Package for Social Science (SPSS) used. A pvalue < 0.05 was considered significant. We could not find a significant correlation between the Q angle values and the patellofemoral indices in all patients (bilateral or only right/left). Within the patients with bilateral patellofemoral pain (n = 15), there was a significant correlation between LPD and PLCI (p = 0.015), LPT and PLCI (p = 0.024) left and LPD and LPT(p = 0.011) right. Similar results were found in patients with pain only on one side. In conclusion, there is no significance between the Q angle and the position of patella. The diagnostic relevance of the Q angle could not be established. PMID- 11482470 TI - The musculocutaneous sural artery flap for soft-tissue coverage after calcaneal fracture. AB - Soft-tissue defects following calcaneal fractures can be covered in a relatively easy and safe procedure. We have modified the familiar distally based sural artery flap by lifting a part of the gastrocnemius muscle. With an inferior pedicle, this musulocutaneous flap can be rotated onto the defect on the sole of the foot and on the heel. Five patients with open fractures of the calcaneus or wound necrosis after osteosynthesis were treated with this procedure. Two defects were covered uneventfully, two flaps were prepared, the rotation being done in a secondary procedure. One patient demanded further revisions, and the flap was partially lost, but the remaining defect was covered after open treatment. This new musculocutaneous sural artery flap can be used for covering even an extensive defect after calcaneal fractures and seems to be a reliable procedure. Morbidity at the donor site is low, and in the case of failure, the free flap remains an alternative. PMID- 11482471 TI - Femoral neck fracture--the cause of death of Emperor Charles IV. AB - We present the oldest documented case of femoral neck fracture of a particular person in the history of orthopaedic surgery. Examination of the skeleton of Charles IV, the King of Bohemia and Roman Emperor living in XIVth century has revealed a fracture of the left femoral neck. This fracture was most probably an indirect cause of his death as it resulted in pneumonia, the immediate cause of death. This fact has been confirmed by contemporary chronicles. PMID- 11482472 TI - Low tibial osteotomy for moderate ankle arthritis. AB - Low tibial osteotomy is one of the significant advances of ankle reconstruction techniques that has been made recently in an effort to halt arthritis in its early stages and leave fusion as the last, not the only, alternative treatment of ankle arthritis. From 1989 to 1995, we performed 18 low tibial osteotomies which included 6 cases of post-traumatic arthritis and 12 cases of degenerative arthritis. The ages of the 7 male and 11 female patients ranged from 18 to 78 years with an average of 41.9 years. The follow-up period lasted a mean of 47.7 months, ranging from 25 to 82 months. The average functional score changed from 49.6 pre-operatively to 88.5 at the last follow up, and showed yearly improvement. Complications included one case of late infection and two cases of implant failure, none of which led to nonunion. The indication for low tibial osteotomy is the intermediate stage of moderate ankle arthritis with a medial joint lesion and intact lateral facet. Using pressure redistribution on the joint surface, this procedure is an alternative treatment for ankle arthritis which may save an arthritic ankle from the fate of fusion or at least postpone fusion surgery. PMID- 11482473 TI - Treatment and clinical management of recurrent knee empyema due to methicillin resistant Staphylococcus aureus (MRSA) after total knee replacement. AB - We describe a patient with recurrent knee empyema and associated complications caused by methicillin-resistant Staphylococcus aureus (MRSA) after total knee replacement and the difficulties and limited options in its clinical management. PMID- 11482474 TI - Non-union of the capitate with associated triangular fibrocartilage tear. AB - We report a case of a 19-year-old boy who presented with a painful wrist 3 months after a fall. Plain radiographs demonstrated a non-union of a capitate fracture which was unrecognised at the time of injury. Magnetic resonance imaging confirmed the diagnosis and also demonstrated a tear of the triangular fibrocartilage complex. This combination of injuries has not been previously described. PMID- 11482475 TI - Peritoneal dialysis should be the first choice of initial renal replacement therapy for more patients with end-stage renal disease. PMID- 11482476 TI - Declining utilization of peritoneal dialysis: time to stop imposing our biases on the patients and let them be dialyzed with the modality of their choice. PMID- 11482477 TI - Development of an implantable artificial lung: challenges and progress. AB - Unlike dialysis, which functions as a bridge to renal transplantation, or a ventricular assist device, which serves as a bridge to cardiac transplantation, no suitable bridge to lung transplantation exists. Our goal is to design and build an ambulatory artificial lung that can be perfused entirely by the right ventricle and completely support the metabolic O2 and CO2 requirements of an adult. Such a device could realize a substantial clinical impact as a bridge to lung transplantation, as a support device immediately post-lung transplant, and as a rescue and/or supplement to mechanical ventilation during the treatment of severe respiratory failure. Research on the artificial lung has focused on the design, mode of attachment to the pulmonary circulation, and intracorporeal versus paracorporeal placement of the device. PMID- 11482478 TI - Continuous venovenous renal replacement therapy using a conventional infusion pump. AB - To evaluate continuous venovenous hemofiltration and hemodiafiltration with a conventional infusion pump in a pediatric sized animal model. Fourteen Maryland pigs weighing 8 to 13 kg were used. A conventional infusion pump (IVAC 571), with a flow of 900 ml/h and a pediatric hemofilter of 0.22 m2 were used. Ringer's solution was used for both the dialysate and the replacement fluid. Each experiment included 1 hour of hemofiltration and 1 hour of hemodiafiltration. Heart rate, arterial blood pressure, pH, Na, K, Cl, and hematocrit were measured every 30 minutes. Mean ultrafiltrate flow was 249.7 +/- 100.3 ml/hr, 240.5 +/- 109.5 ml/hr with hemofiltration and 271 +/- 101.1 ml/hr with hemodiafiltration, the differences not being significant. No significant changes were seen in heart rate, blood pressure, hematocrit, electrolytes, or pH. Pressure in the circuit rose from 107.7 +/- 70.3 mm Hg at the beginning of the experiment to 234.2 +/- 118.1 mm Hg after 2 hours (p < 0.05). The technique was well tolerated by all the pigs. Continuous venovenous hemofiltration and hemodiafiltration with a conventional infusion pump is a possible alternative to conventional methods of extrarenal replacement therapy in neonates and infants. PMID- 11482479 TI - Pretreatment of a Dacron graft with tissue factor pathway inhibitor decreases thrombogenicity and neointimal thickness: a preliminary animal study. AB - The purpose of this study was to evaluate the effects of locally applied TFPI on the reduction of neointimal thickness in Dacron grafts. Seven millimeter internal diameter 5 cm lengths of albumin coated knitted Dacron grafts were interposed in the infrarenal aorta in 14 mongrel dogs. Before implantation, the grafts were immersed in saline solution containing 200 microg/ml of TFPI (TFPI group, n = 7) or 100 IU/ml of heparin (control group, n = 7) for 15 minutes at room temperature. Three months after implantation, neointimal thickness and percentage of graft stenosis were measured by computerized planimetry. All grafts were patent 3 months after implantation. Thrombus was found in one graft in the TFPI group, but observed in three of seven control grafts. Neointimal thickness in the TFPI group was significantly smaller than that in the control group (mean +/- SD, 0.26 +/- 0.1 mm vs. 0.57 +/- 0.15 mm, p < 0.001). Percentage of graft stenosis was significantly lower in the TFPI group than in the control group (13.4 +/- 5.3% vs. 26.9 +/- 7.0%, p < 0.001). Scanning electron micrographs showed that the neointima of TFPI treated grafts were completely covered by endothelial cells. The present results indicate that locally applied TFPI reduces thrombogenicity and neointimal thickness in albumin coated knitted Dacron grafts. PMID- 11482480 TI - End-to-side anastomosis using a new sutureless vascular prosthesis. AB - Performing a conventional vascular anastomosis during minimally invasive surgery remains a technically difficult and time-consuming task. The purpose of our study was to assess the efficacy of a new sutureless vascular prosthesis. Through a left thoracotomy, a short segment of the descending aorta was bypassed in eight sheep. Our Dacron prosthesis had a cuff at one extremity. The cuff was introduced into the artery through an arteriotomy and attached to the arterial wall with clips. Surgery was performed on the sheep again 12-15 weeks later, and the prosthesis was retrieved for macroscopic and microscopic examination. At surgery, insertion of the prosthesis was easy, fast, and safe. A 3-4 mm space between each clip was required for proper attachment. At the second surgery, all prostheses were completely patent and there were no anastomotic stenoses or aneurysms. This new sutureless vascular prosthesis was easy and safe in its placement and might prove useful for video assisted vascular aortic surgery. PMID- 11482481 TI - Preliminary experience with a cuffed ePTFE graft for hemodialysis vascular access. AB - The purpose of this study was to determine graft patency and blood flow rates in recipients of a new cuffed ePTFE graft (Venaflo graft) used for hemodialysis access. A pilot study was conducted with 12 (7 men, 5 women) consecutive patients (age range, 36-76 yr; mean, 65 yr). All patients were recipients of a new cuffed PTFE graft placed for hemodialysis access. Seven were high risk because of a prior history of clotted hemodialysis accesses (1-6; mean, 3.3). Blood flow rates were determined by ultrasound dilution technique at 3 month intervals. One year and 2 year overall graft patency rates were 90.9% and 68.2%, respectively. One graft (high risk, six prior grafts) was lost to thrombosis in the first year; two grafts (one high risk, four prior grafts) were lost to thrombosis in the second year of follow-up. No graft thrombosis resulted from stenosis at the graft-vein anastomosis. Blood flow rates ranged from 550 to 2,110 ml/min (mean, 1,086 ml/min; n = 8) when first measured 3 months after graft placement. Similar flow rates were observed at 12 months (mean, 1,043 ml/min; n = 7) and 24 months (mean, 1,014 ml/min; n = 4) in grafts available for comparison. Dialysis flow rates in excess of 350 ml/min were possible with all patent grafts. A cuffed ePTFE graft provided stable blood flow and satisfactory graft patency during 2 years of follow-up, even in high risk patients with a prior history of vascular access thrombosis. PMID- 11482482 TI - Newly developed drug delivery system in cancer chemotherapy using direct electric current. AB - To improve the antitumor effect of chemotherapy on a target organ, we experimentally investigated pharmacokinetic alteration of methotrexate (MTX) induced by an electric current in rats to ascertain whether the local concentration of MTX could be enhanced while reducing the concentration in blood. Six male Wistar rats had platinum electrodes introduced into both kidneys. Three rats were subjected to direct current (3.5 V, 50 microA) for 3 hours, while another three control rats received no current. While providing the current to the treated group, all of the rats were intravenously injected with MTX at a dose of 1 mg/kg. The MTX concentration in urine (excreted from each ureter) and serum was measured by fluorescence polarization immunoassay. Cumulative MTX renal excretion rate was significantly higher in the cathode than anode side in the treated group (p < 0.01). Serum MTX concentration at 0 to 5 min was significantly reduced in the treated group (p < 0.03). The first part of the area under the curve (i.e., alpha curve) was markedly increased in the treated group versus controls (p = 0.05). Electric therapy using a slight direct current could gather anionized MTX to the cathode, while reducing MTX concentration in the serum of experimental rats. PMID- 11482483 TI - Successful repair of esophageal injury using an elastin based biomaterial patch. AB - An esophageal injury with significant tissue loss is very difficult to repair. We conducted an in vivo study to test our elastin based acellular biomaterial patch to repair such defect. The patch was made from porcine aorta, by decellularization and sterilization. Collagen fibers were preserved to retain mechanical strength and enhance cellular in-growth. Ten domestic pigs underwent right thoracotomy. A 2 cm circular defect was made on the distal esophagus, excising half its circumference, and was repaired using the biomaterial patch and sutures. Soon after the procedure, the animals resumed oral feeding. They were followed for clinical status, weight gain, barium studies, and endoscopic studies, and were killed after 6 weeks to 4 months. All ten animals survived long term, with a procedure success rate of 100% (10 of 10). With the exception of one pneumothorax, no complications occurred, and all animals resumed oral feeding and gained weight. Endoscopic studies showed mucosal coverage by 6 weeks, with minimal stricture at the repair site. Excised specimens showed complete mucosal coverage with regeneration of all three layers. Our biomaterial patch can be used safely and reliably for repair of esophageal injury with significant tissue loss when repaired immediately as in our experiment. PMID- 11482484 TI - Development of an implantable artificial anal sphincter by the use of the shape memory alloy. AB - In this study, we developed and assessed an artificial anal sphincter driven by an shape memory alloy actuator (AS-SMA). The performance characteristics of the device were analyzed with a measurement system. Assessment showed that the AS-SMA could generate a pressure of 55 mm Hg at an atmospheric temperature of 36 degrees C, and displacement of the SMA actuator was 7.5 mm when the temperature of the SMA plate was 55 degrees C. To evaluate opening and closing, we studied a piglet colostomy model, in which the AS-SMA was implanted around the colostomy in the extraperitoneal space. Flow control tests using living porcine intestine revealed that the AS-SMA could maintain fecal continence against an intestinal pressure of 75 mm Hg. The high pressure zone corresponding to the location of the device was demonstrated in a manometric examination. For 6 days after surgery, we activated the AS-SMA twice a day and observed the bowel movements. The animal experiment indicated that the AS-SMA is able to control the bowel movements of patients with fecal incontinence if several problems, such as burning of tissue around the device and compression injury of the intestine, are resolved. PMID- 11482485 TI - Influence of static pressure and shear rate on hemolysis of red blood cells. AB - The purpose of this study was to investigate the effect of multiple mechanical forces in hemolysis. Specific attention is focused on the effects of shear and pressure. An experimental apparatus consisting of a rotational viscometer, compression chamber, and heat exchanger was prepared to apply multiple mechanical forces to a blood sample. The rotational viscometer, in which bovine blood was subjected to shear rates of 0, 500, 1,000, and 1,500 s(-1), was set in the compression chamber and pressurized with an air compressor at 0, 200, 400, and 600 mm Hg. The blood temperature was maintained at 21 degrees C and 28 degrees C. Free hemoglobin at 600 mm Hg was observed to be approximately four times higher than at 0 mm Hg for a shear rate of 1,500 s(-1) (p < 0.05). The results suggest that the increase in hemolysis is strongly related to pressure when high shear rates are applied to the erythrocytes. The data acquired in this study will be helpful in the development of artificial organs, where it will facilitate the prediction of hemolysis in flow dynamics analysis, flow visualization, and computational fluid dynamics. PMID- 11482486 TI - Effects of fibrinogen residence time and shear rate on the morphology and procoagulant activity of human platelets adherent to polymeric biomaterials. AB - Fibrinogen readily adsorbs to the surface of biomaterials and, because of its demonstrated ability to support platelet adhesion and aggregation, plays a role in thrombotic events associated with the implantation of synthetic materials in the human body. Thus, understanding the factors influencing the interactions of fibrinogen with biomaterials, and how platelet responses are affected, is crucial for the development of synthetic materials exhibiting improved blood compatibility. In this study, the effects of fibrinogen residence time and shear rate on the procoagulant activity of adherent platelets, along with their morphologic status, as deduced from scanning electron microscopy, were investigated. To examine whether adherent platelets promoted the generation of thrombin, polymeric materials (polytetrafluoroethylene, polyethylene, and silicone rubber) preadsorbed with fibrinogen were exposed to platelet suspensions at different wall shear rates and then incubated with clotting factors for 5 minutes under static conditions. The amount of thrombin generated per platelet was calculated from the optical density of the color developed by adding substrate S-2238. Scanning electron microscopy images of the platelets revealed that the platelets exhibited different morphologies, depending on the shear rate and residence time of the adsorbed fibrinogen. Platelets ranged from their normal discoid shape observed primarily under static conditions, to that of fully spread platelets. Results from this study show that platelets, in the presence of shear forces, undergo activation on exposure to surfaces on which adsorbed fibrinogen has resided for short residence times rather than long residence times. Interestingly, studies examining the procoagulant responses of such adherent platelets demonstrated that the platelets attached to the fibrinogen coated materials did not promote significant thrombin generation. Such low prothrombinase activity of adherent platelets suggests that adsorbed fibrinogen, while capable of supporting platelet adhesion and spreading on biomaterials, does not necessarily enhance the procoagulant activity of adherent platelets. PMID- 11482487 TI - Platelet adhesion to heparin coated oxygenator fibers under in vitro static conditions: impact of temperature. AB - Heparin coating of cardiopulmonary bypass (CPB) circuitry may attenuate the platelet consumption associated with CPB. We investigated the effect of temperature on the interaction between platelet and heparin coated surfaces under in vitro static conditions. Heparin coated and non coated oxygenator fibers were incubated with heparinized whole blood at 37 degrees C and 22 degrees C. The incubation time was set at 30, 60, 180, and 300 minutes. The number of platelets adhering to each fiber was assessed with enzyme immunoassay using monoclonal antibody against platelet receptor protein CD 61(GPIIbIIIa). As an index of platelet activation, plasma soluble(s) P-selectin levels were measured by enzyme linked immunosorbent assay. Under normothermia, the number of adherent platelets on the non coated surface increased significantly after 300 min of incubation. Platelet adhesion was reduced significantly by heparin coating of the surface and was kept constant after 300 min. Under hypothermia, heparin coating was also associated with significant reduction of platelet adhesion. The levels of sP selectin did not correlate with the extent of platelet adhesion. Our results suggest that heparin coating is effective in decreasing platelet adhesion to the synthetic surface tested regardless of the temperature under static conditions. Inhibition of platelet activation on the heparin coated surface may be masked by standard dose heparinization. PMID- 11482488 TI - Interleukin-8, aquaporin-1, and inducible nitric oxide synthase in smoke and burn injured sheep treated with percutaneous carbon dioxide removal. AB - We previously showed that a percutaneous arteriovenous gas exchanger was effective in removing CO2 and reversing respiratory failure in an ovine model of adult respiratory distress syndrome (ARDS) produced by smoke inhalation and burn injury (Alpard et al., Ann Surg 230:215-224, 1999). In this study, we tested the hypothesis that arteriovenous CO2 removal (AVCO2R) lessened endogenous inflammation in the lung. Myeloperoxidase activity, aquaporin-1 (AQP-1), interleukin-8 (IL-8), and inducible nitric oxide synthase mRNAs as well as aquaporin-1, and IL-8 protein were measured in ovine lung tissue. Lung tissue was taken at 96 h (time of sacrifice) from animals with combined smoke inhalation and 40% third degree dermal burn and subsequently treated with AVCO2R or sham (ventilator alone) after onset of ARDS (PaO2:FiO2 ratio of < 200). Myeloperoxidase activity was 1.862 +/- 0.302 U/mg protein in the ventilator group and 0.830 +/- 0.141 in the AVCO2R plus ventilator group. AQP-1 mRNA was 140,482 +/- 31,702 copies/microg total RNA in the ventilator group and 61,854 +/- 22,433 copies/microg total RNA in the AVCO2R plus ventilator group (p = 0.076). mRNA for IL-8 mRNA in the ventilator alone treated animals was 74,000 +/- 3,300 copies/microg total RNA compared to < 1,000 copies/microg total RNA in the ventilator plus AVCO2R group. This result was highly significant (p < 0.001) Inducible nitric oxide synthase mRNA was 7,853 +/- 2,229 copies/microg total RNA for the AVCO2R group and 5,854 +/- 2,070 copies/microg total RNA for the ventilator managed animals. These differences were not statistically significant (p = 0.54). Percutaneous AVCO2R produced a specific decrease in IL-8 in the smoke and burn injured animals. Furthermore, this effect was consistent with cell signaling mechanisms that increase the expression of IL-8 by cyclic stretching and the observed reduction in the number of neutrophils in the lung parenchyma. Therefore, we speculate that the mechanism by which CO2 removal exerts a beneficial effect may be due to both decreases in ventilatory requirements, with an accompanying reduction in alveolar stretching, and reduction of neutrophil numbers in lung tissue. PMID- 11482489 TI - Recirculation in double lumen catheter veno-venous extracorporeal membrane oxygenation measured by an ultrasound dilution technique. AB - Recirculation is a limiting factor for oxygen delivery in double lumen catheter veno-venous extracorporeal membrane oxygenation (DLVV-ECMO). This study compares three different methods for the determination of the recirculation fraction during double lumen catheter veno-venous ECMO at ECMO flow rates of 150, 125, 100, 75, and 50 ml/kg.min in nine lambs: (1) an ultrasound dilution method, in which the change in ultrasound velocity in blood after injection of a saline bolus as a marker is used for determination of recirculation; (2) an SvO2 method using real mixed venous blood oxygen saturation, the gold standard, for determination of recirculation fraction; and (3) the CVL method, in which oxygen saturation of a blood sample of the inferior vena cava is considered to represent mixed venous oxygen saturation. In all methods, the recirculation fraction increased with increasing ECMO flow rate. The correlation coefficient between the ultrasound dilution method and the SvO2 method was 0.68 (p < 0.01); mean difference was -2.4% (p = 0.6). Correlation coefficient between the ultrasound dilution method and the CVL method was 0.48 (p < 0.01); mean difference was 18.1% (p < 0.01). The correlation coefficient between the SvO2 method and the CVL method was 0.51 (p < 0.01); mean difference was -15.7% (p < 0.01). The ultrasound dilution method is a useful method for measurement of the recirculation fraction in DLVV-ECMO and is easier to use than the other methods. PMID- 11482490 TI - Model based sensitivity analysis of arterial pressure response to hemodialysis induced hypovolemia. AB - The role of hemodynamic and regulatory factors in the arterial pressure response to hemodialysis induced hypovolemia was investigated by means of a computer model of the cardiovascular system, including the main short-term pressure regulatory mechanisms. The model mimics the arterial and venous systemic circulation, Starling's law and inotropic heart regulation, arterial and cardiopulmonary baroreflex controls of resistance, and capacitance vessels. All of the model parameters have a clear physiologic meaning: 10 represent the systemic circulation, 4 describe cardiac pump performance, and 3 characterize baroreflex regulation. Sensitivity analysis is performed to determine the effect of each parameter on the pressure response to mild hypovolemia (a 10% blood volume reduction after 4 hours). The results demonstrate that circulatory parameters, such as resistances and compliances, have no relevant effect upon the pressure response. Conversely, regulation of venous capacity seems to play a pivotal role in sustaining arterial pressure during hemodialysis induced hypovolemia. Regulation of systemic peripheral resistance exerts a compensatory action only as long as the blood volume reduction is < 5%, but it is inadequate to compensate for a larger blood volume reduction when venous capacity regulation is absent. A paradoxical arterial pressure increase during hypovolemia can be referred to a prevalence of cardiopulmonary afferences in the regulatory process. PMID- 11482491 TI - A new method to control ultrafiltration in conventional continuous renal replacement therapy. AB - Conventional continuous arteriovenous hemofiltration/hemodialysis (CAVH/D) and slow continuous ultrafiltration (SCUF) are types of continuous renal replacement therapy (CRRT) in which the ultrafiltrate (UF) volume is controlled imprecisely with a UF clamp, which is labor intensive, demanding frequent adjustment to preclude excessive fluid removal. We devised a simple method for precise control of the UF volume. Seven CRRTs in the form of CAVH, CAVHD, and SCUF were performed in four patients with massive edema. A standard circuit was created in each case using blood tubing sets and an HF 400 hemofilter obtained from MinnTech. Standard intravenous infusion tubing connected to an infusion pump (IMED, Gemini PC-2 volumetric pump/controller) with its proximal end inserted into the dialysate port at the venous end of the hemofilter, and the distal end draining into a plastic bag, was used to control the UF rate. Dialysis was added to the circuit using another pump connected to the dialysate port at the arterial end of the hemofilter. Treatment time ranged from 27 to 78 hours. Target fluid removal was achieved in all treatments, and the net UF rate required only once daily adjustment for total fluid intake. Mean time to reporting a problem by the intensive care nurse was 30 hours (range, 25-30 hours), and mean time to filter clotting was 38 hours (range, 27-40 hours). This set-up is less labor intensive, more cost effective, and is applicable in areas lacking automated machines. Future development of tubing for UF designed as above may further reduce cost. PMID- 11482492 TI - A durable, non power consumptive, simple seal for rotary blood pumps. AB - One of the key technologic requirements for rotary blood pumps is the sealing of the motor shaft. A mechanical seal, a journal bearing, magnetic coupling, and magnetic suspension have been developed, but they have drawbacks such as wear, thrombus formation, and power consumption. A magnetic fluid seal was developed for an axial flow pump. A magnetic fluid seal is durable, simple, and non power consumptive. Long-term experiments and finite element modeling (FEM) analyses confirmed these advantages. The seal body was composed of a Ned-Fe magnet and two pole pieces; the seal was formed by injecting ferrofluid into the gap (50 microm) between the pole pieces and the motor shaft. To contain the ferrofluid in the seal and to minimize the possibility of ferrofluid making contact with blood, a shield with a small cavity was attached to the pole piece. While submerged in blood, the sealing pressure of the seal was measured and found to be 188 mm Hg with ferrofluid LS-40 (saturated magnetization, 24.3 kA/m) at a motor speed of 10,000 rpm and 225 mm Hg under static conditions. The magnetic fluid seals performed perfectly at a pressure of 100 mm Hg for 594 + days in a static condition, and 51, 39+, and 34+ days at a motor speed of 8,000 rpm. FEM analyses indicated a theoretical sealing pressure of 260 mm Hg. The state of the magnetic fluid in the seal in water was observed with a microscope. Neither splashing of magnetic fluid nor mixing of the magnetic fluid and water was observed. The specially designed magnetic fluid seal for keeping liquids out is useful for axial flow blood pumps. The magnetic fluid seal was incorporated into an intracardiac axial flow pump. PMID- 11482493 TI - Pitfalls in the development of a rotary blood pump controller. AB - The controller presents a major obstacle in the development of the rotary blood pump as a left ventricular assist device (LVAD). Clinically, LVAD flow is a good indicator in the regulation of circulatory conditions and pump flow changes, depending on pump preload and afterload. Many investigators have tried estimating pump flow by referencing the motor current. There have been pitfalls in in vitro experimental settings, however. Using a test loop with a pneumatically driven LV chamber and a centrifugal pump as an LVAD, we monitored pump flow and pressure head to evaluate the pump performance curve (H-Q curve). Under pulsatile LV conditions, the H-Q curve was a loop that changed, depending on LV contractility. The pneumatically driven LV chamber cannot mimic the Starling phenomenon, so the developed LV pressure does not change according to the LV preload. Rotary pump flow estimation is the most effective control method. In pulsatile conditions, however, the H-Q curve is a loop that changes under various LV contractility conditions, complicating determination of linear equation for calculating flow. In addition, the LV chamber in the test loop cannot mimic native heart contractility as described by Starling's law. This finding can lead to a misanalysis of the H-Q curve under pulsatile conditions. PMID- 11482494 TI - HeartMate III: pump design for a centrifugal LVAD with a magnetically levitated rotor. AB - A long-term, compact left ventricular assist device (LVAD), the HeartMate III, has been designed and fabricated, featuring a centrifugal pump with a magnetically levitated rotor. The pump has been optimized by in vitro testing to achieve a design point of 7 L/min against 135 mm Hg at high hydrodynamic efficiency (30%) and to be capable of up to 10 L/min under such a load. Furthermore, the pump has demonstrated no mechanical failures, low hemolysis (4 10 mg/dl plasma free Hb), and low thrombogenicity during six (40, 27, 59, 42, 27, and 49-day) in vivo bovine studies. PMID- 11482495 TI - Particle image velocimetry measurements of blood velocity in a continuous flow ventricular assist device. AB - The third prototype of a continuous flow ventricular assist device (CFVAD3) is being developed and tested for implantation in humans. The blood in the pump flows through a fully shrouded four-bladed impeller (supported by magnetic bearings) and through small clearance regions on either side of the impeller. Measurements of velocities using particle image velocimetry of a fluid with the same viscosity as blood have been made in one of these clearance regions. Particle image velocimetry is a technique that measures the instantaneous velocity field within an illuminated plane of the fluid field by scattering light from particles added to the fluid. These measurements have been used to improve understanding of the fluid dynamics within these critical regions, which are possible locations of both high shear and stagnation, both of which are to be avoided in a blood pump. Computational models of the pump exist and these models are currently being used to aid in the design of future prototypes. Among other things, these models are used to predict the potential for hemolysis and thrombosis. Measurements of steady flow at two operating speeds and flow rates are presented. The measurements are compared with the computed solutions to validate and refine, where necessary, the existing computational models. PMID- 11482496 TI - A novel miniature ventricular assist device for hemodynamic support. AB - The HemoDynamics Systems enabler is a new cardiac assist pump that can expel blood from the left ventricle and provide pulsatile flow in the aorta. We evaluated the efficacy of the 18 Fr enabler. The enabler was inserted from the left ventricular apex into the ascending aorta in eight sheep. Heart failure (mild, moderate, and severe) was induced by microsphere injection into the coronary arteries to reduce cardiac output by 10-30%, 31-50%, and more than 50% from baseline, respectively. The enabler was activated, and its flow was increased to approximately 2.0 L/min. Hemodynamic variables were recorded before and after activation. In moderate heart failure, cardiac output and mean aortic pressure increased from 2.3 +/- 0.6 L/min and 59 +/- 12 mm Hg before assist to 2.8 +/- 0.6 L/min and 70 +/- 8 mm Hg at 30 minutes after activation, respectively (p < 0.01). Left atrial pressure decreased from 17 +/- 3 to 13 +/- 4 mm Hg (p < 0.05). Similar findings were observed in mild and severe heart failure. Despite its small diameter, the enabler significantly improved the hemodynamics of failing hearts and may potentially serve as a means of peripheral left ventricular support. Further study is warranted. PMID- 11482497 TI - Near infrared spectrophotometry reflects cerebral metabolism during hypothermic circulatory arrest in adults. AB - Near-infrared spectrophotometry (NIRS) is assumed to reflect cerebral oxygenation during hypothermic circulatory arrest (HCA). However, the rationale for the use of NIRS as a marker of cerebral metabolism remains to be elucidated. We examined whether cerebral oxygenation measured by NIRS correlated with cerebral metabolic function assessed by cerebral oxygen extraction ratio (OER) during HCA in adults. NIRS was continuously monitored in 14 patients undergoing thoracic aortic surgery using HCA (17.9 +/- 2.9 degrees C esophageal temperature at HCA onset). Jugular venous oxygen saturation and OER were also monitored. OER was significantly reduced from 37.3 +/- 7.2% at the onset of cardiopulmonary bypass to 13.6 +/- 6.2% at the onset of HCA (p < 0.0001). A linear decrease in oxygenated-hemoglobin (oxy-Hb) and increase in deoxygenated-hemoglobin (deoxy-Hb) were found during HCA, which returned to baseline levels after rewarming. The rate of decrease in oxy-Hb and increase in deoxy-Hb were -0.63 +/- 0.45 and 0.51 +/- 0.30 (10(-5) OD/cm per sec), respectively, both of which significantly correlated with OER at the onset of HCA (R2=0.739 and 0.633; p < 0.0001 and p = 0.0007, respectively). NIRS may serve as a reliable diagnostic modality for monitoring cerebral metabolism during aortic surgery using HCA. PMID- 11482498 TI - Hemodialysis for removal of iodinated contrast media. PMID- 11482499 TI - Treatment of HCV-related liver disease in the dialysis population: a novel challenge for clinical nephrologists. PMID- 11482500 TI - Daily dialysis Kt/V and flexible schedules: is it possible to control efficiency, when and how? AB - BACKGROUND: Daily hemodialysis is a promising treatment schedule but uniform criteria for defining efficiency are lacking. METHODS: On our daily dialysis (DD) schedule, duration is flexible (2-3 hours, patients are free to add up to 30 min/session), Qb 250-350 mL/min; dialyser 1.6-1.8 m2. Study was performed on 12 pts on DD for > or = 2 months, with > or = 4 Kt/V on subsequent days, tested in the same laboratory. GOAL: To evaluate variability and identify a simple method for weekly calculation, Kt/V was assessed for 133 sessions. RESULTS: On flexible DD, variability of Kt/V-session is high (relative error 4.9%-22%). On flexible schedules, within the time range chosen (2-3 hours) variability of average hourly Kt/V is lower (standard deviation: min (0.014; max (0.052 hour, relative error 4.9%-10%) allowing calculation of weekly Kt/V (averaging 3 sessions: relative error < 6%) suitable for clinical practice. CONCLUSIONS: Flexible schedules, allowing patients to increase treatment time, are an interesting clinical option, but a challenge for Kt/V assessment. PMID- 11482501 TI - Renal replacement therapy: physical properties of hollow fibers influence efficiency. AB - INTRODUCTION: Physical properties of filters for continous renal replacement therapy have a great impact on biocompatibility. According to Poiseuille's law, a filter with more and shorter hollow fibers should offer a decreased pressure drop and, therefore, lower transmembrane pressure (TMP). The aim of this study was to study the effect of a new filter configuration in terms of TMP and clotting compared with the standard configuration. METHODS: In a prospective randomized cross-over study 2 polysulphone hollow fiber hemofilters, one handmade, which differed only in length and number of hollow fibers were compared. In each group 12 filters were investigated during continous venovenous hemofiltration in patients with acute renal failure due to septic shock. Pressures were measured every 3 hours and running time until filter clotting was documented. Mediators before and after the filter, at the end of treatment and in filtrate were assessed. RESULTS: The standard filter with longer hollow fibers had significantly lower TMPs (106 vs. 194 mmHg, p=0.02) and longer running times (1276 vs. 851 min, p=0.04). There were no differences in hematocrit, total protein, cellular and plasmatic coagulation or blood temperature. No significant elimination of mediators was shown. CONCLUSION: In contrast to our expectations, the filter with the longer hollow fibers had a better performance, as it ran longer and had lower TMP This may be due to slower blood flow leading to an increase in blood viscosity in a filter with a larger cross section. PMID- 11482502 TI - Influence of recombinant human erythropoietin therapy on plasma endothelin-1 levels during hemodialysis. AB - The correction of anemia with human recombinant erythropoietin (rHuEPO) in end stage renal disease is associated with hypertension in about one third of hemodialysis patients. The pathogenesis of the rHuEPO-induced hypertension is still uncertain, though evidence of the involvement of endothelial cells has emerged. The aim of this study was to determine plasma endothelin-1 during hemodialysis and to compare the endothelin-1 levels in hemodialysis patients with and without rHuEPO substitution. Nineteen stable patients (13 male and 6 female, mean age 62 +/- 11 years) with end stage renal disease were studied. Cuprophan dialysers (GFS 12, Gambro, Lund, Sweden) were used for hemodialysis in all cases. rHuEPO (40 U/kg s.c.) was administered to 10 patients. Blood pressure (BP; RR mmHg) and blood volume changes (deltaBV; hemoglobinometry %) were serially measured. Samples were taken before and every hour during hemodialysis. Plasma endothelin-1 was measured by ELISA (R&D Systems, Minneapolis, USA) and corrected for hemoconcentration. Endothelin-1 concentration was elevated before commencement of hemodialysis (1.16 +/- 0.36 pg/ml) when compared to healthy controls (ref. 0.3-0.9) and increased to 1.47 +/- 0.51 pg/ml by the end of the session (p<0.05). In patients under rHuEPO-substitution plasma endothelin-1 was higher when compared to patients without substitution before (1.25 +/- 0.3 vs. 1.05 +/- 0.3 pg/ml) and at the end of HD (1.62 +/- 0.5 vs. 1.28 +/- 0.3 pg/ml, p<0.05). There was no difference in BP and deltaBV between the two groups during treatment. Plasma endothelin-1 was higher in hemodialysis patients and there was a continuous rise in plasma endothelin-1 during a session. Comparison of two groups of hemodialysis patients with and without s.c. rHuEPO-replacement treatment revealed a significantly higher plasma endothelin-1 concentration in patients with s.c. rHuEPO treatment. However, the elevated endothelin-1 levels were not accompanied by arterial hypertension. PMID- 11482503 TI - Interrelationship between cardiac structure and function and incidence of arrhythmia in peritoneal dialysis patients. AB - Cardiovascular complications in patients with end-stage renal disease requiring dialytic therapy are frequent and account for approximately 40% of all deaths in these patients. The aim of this study was to analyze the occurrence of cardiac arrhythmia in peritoneal dialysis (PD) patients with respect to the changes in left ventricular structure and function. To determine characteristics of arrhythmia in patients on PD for chronic renal failure, 30 patients (18 male and 12 female; aged 54.1+/-13.8 years) underwent twice (interval of 20+/-4.1 months) ambulatory 24 hour Holter ECG monitoring. At the same time all the patients were analyzed by echocardiography and pulsed Doppler echocardiography to estimate cardiac structure and function. Ventricular arrhythmias were seen in 9 patients (30%) during the first examination and in 13 patients (43.3%) on the second. Ventricular arrhythmias were observed only in patients with left ventricular hypertrophy (LVH). Supraventricular arrhythmias were seen in 12 (40%) and 17 (56.7%) patients. The majority of these patients also had LVH, with 11/12 (91.7%) patients at the first examination and 15/17 (88.2%) at the second respectively. We conclude that the incidence of arrhythmia is primarily dependent on the presence of LVH in PD patients. It appears that peritoneal dialysis does not provoke or aggravate arrhythmia. PMID- 11482504 TI - Mono and bi-ventricular assistance: their effect on ventricular energetics. AB - When mono- and bi-ventricular mechanical assistance is used for heart recovery, its control strategy and circulatory variables affect ventricular energetics (external work-EW, oxygen consumption-VO2, cardiac mechanical efficiency-CME). This study is based on the data obtained in vitro and presents an analysis of the effects of the mono- and bi-ventricular mechanical assistance on ventricular energetics. The assistance was conducted on the principle of counterpulsation with atrio-arterial connection. It includes the following stages: 1) the characterisation of the isolated ventricle model in terms of EW, VO2 and CME as a function of the filling pressure and peripheral resistance, 2) modelling of left ventricular and pulmonary dysfunction, followed by left ventricular and bi ventricular assistance. Experimental data enable us to draw the following conclusions: * in general, the greatest hemodynamic improvement does not correspond to the highest energetic improvement, * LVAD assistance deteriorates left ventricular CME while its effect on right ventricular energetics depends on the value of right ventricular elastance (Emax). Right ventricular CME is deteriorated by BVAD assistance irrespective of right Emax, * the energetics optimisation in bi-ventricular assistance is closely related to the right Emax, which could probably be a deciding factor in the choice of the assistance mode. PMID- 11482505 TI - Comparison of pig, human and rat hepatocytes as a source of liver specific metabolic functions in culture systems--implications for use in bioartificial liver devices. AB - The limited availability of human hepatocytes results in the use of animal cells in most bioartificial liver support devices. In the present work, clinically relevant liver specific metabolic activities were compared in rat, pig and human hepatocytes cultured on liver-derived biomatrix to optimize the expression of differentiated functions. Pig hepatocytes showed higher rates of diazepam metabolism (2.549+/-0.821 microg/h/million cells vs. 0.474+/-0.079 microg/h/million cells rats, p<0.005, and vs. 0.704+/-0.171 microg/h/million cells in man, p<0.005) and of bilirubin conjugation (21.60116+/-8.433237 micromoles/l/24 h vs. 6.786809+/-2.983758 in man, p<0.001 and vs. 9.956538+/ 1.781016 micromoles/l/24 h in rats, p<0.005). Urea synthesis was similar in pig and in human hepatocytes (150+/-46.3 vs. 144.8+/-21.46 nmoles/h/million cells) and it was lower in rats (84.38+/-35.2; p<0.001 vs. man, p<0.02 vs. pig). High liver specific metabolic activities in cultured pig hepatocytes further support their use as a substitue for human cells in bioartificial liver devices. PMID- 11482506 TI - Ethylene glycol dimethacrylate cross-linking anion exchange resin as phosphate binder: effects on rat gut and digestion by small intestine contents. AB - In the present study, ethylene glycol dimethacrylate cross-linking 4 vinylpyridinium anion exchange resin (EGDMA-4VP) effectively bound dietary phosphate in normal rats. However, EGDMA-4VP induced more adverse effects in rat gut than cellulose or Dowex 1X2 (both of which have higher water content), and caused damage to the intestine. In order to resolve this seeming paradox, digestion of EGDMA-4VP with rat small intestine content (S-9 fraction) and carboxyl esterase was investigated in vitro to examine the stability of the resin under conditions it would be subjected to as an orally administered medicine. EGDMA-4VP was digested by small intestinal enzymes, with the exception of carboxyl esterase, and the degradation product ethylene glycol (EG) caused reversible relaxation of longitudinal muscle (but not circular muscle) in rat small intestine. Degradation products increased uptake of 3H2O into primary cultured rat small intestinal muscle cells, but the increase was not significant. PMID- 11482507 TI - Privacy. PMID- 11482508 TI - Techniques for stimulating spinal fusion: efficacy of electricity, ultrasound, and biologic factors in achieving fusion. AB - Many advancements have been made in an attempt to decrease the rate of pseudoarthrosis. Some of these have involved the actual fusion surgery. Fusions with internal fixation devices are able to more effectively eliminate motion during the healing process, thus leading to increased fusion rates. Electrical stimulation and ultrasound were initially developed to aid in fracture healing, but have shown to be efficacious in spinal fusion, as well. Biologic growth factors have long been known to control the bone growth process. These proteins have been identified and isolated for use in augmenting spinal fusion. The discovery of bone morphogenetic protein has great promise in significantly improving fusion rates over the use of either allograft or autograft. PMID- 11482509 TI - Coralline hydroxyapatite as bone substitute in orthopedic oncology. AB - We retrospectively reviewed the results of 71 consecutive patients managed with coralline hydroxyapatite (Pro-Osteon 500; Interpore Cross International) for bone defects produced as a result of surgical extirpation of bone tumors. All patients underwent a thorough physical examination and a radiographic evaluation at baseline and follow-up by the same senior surgeon (R.B.I.). Seventy-one patients were followed up for an average of 2.4 years (range, 8.1 months to 3.5 years); mean age of female patients was 39.8 years and that of male patients was 30 years. The functional status of patients at follow-up was evaluated based on comparison with preoperative functional status. Radiographic evaluation was performed and graded as follows: stage I (obvious margins), stage II (hazy margins), and stage III (obvious incorporation). The average time of return of preoperative function was 7.6 weeks. There were 3 major and 9 minor complications. We conclude that coralline hydroxyapatite graft in the form of Pro Osteon 500 is a viable option for the management of bone defects in orthopedic oncology. PMID- 11482510 TI - Ilioinguinal approach to manage benign pelvic and acetabular tumors. AB - Benign osteoblastic and cystic lesions involving the anterior column and wall of the pelvis and acetabulum are technically difficult areas to approach and achieve successful surgical outcomes. Tumors in this area are rare, leaving little information in the literature regarding how best to surgically approach them. Previous authors have reported small series of patients treated using anteromedial, iliofemoral, and intracapsular dissections. The ilioinguinal approach has been extensively described and used in the modern-day treatment of pelvic and acetabular fractures involving the anterior column and wall. Despite its use for acetabular trauma, application of the ilioinguinal approach in the treatment of benign pelvic and acetabular lesions has not been described. We reviewed the management of 5 patients with benign pelvic or acetabular tumors treated using the ilioinguinal approach. Diagnoses included aneurysmal bone cysts in 3 patients and osteoid osteomas in 2 patients, involving the pelvic anterior column and wall. The approach has permitted satisfactory exposure of the lesions for operative treatment. At an average follow-up of 3 years (range, 2-4 years), excellent functional outcome and no recurrences were noted. There were 2 patients with lateral femoral cutaneous nerve palsies, both of which resolved completely by 8 weeks. Encouraging results from our study demonstrate a new application for the ilioinguinal approach in the treatment of benign anterior column and wall lesions of the pelvis and acetabulum. PMID- 11482511 TI - Gouty tenosynovitis of the wrist. AB - A 56-year-old man presented with a mass of the right wrist limiting excursion of the digits. Operative exploration demonstrated gouty infiltration of the flexor tendons. The tendons were debrided of the urate crystalline material and the motion of the digits returned, preserving all tendon function. The patient later returned with symptoms of carpal tunnel in the contralateral wrist. Exploration of the carpal tunnel revealed similar infiltration of the flexor tendons. The tendons were debrided and the carpal tunnel syndrome resolved. PMID- 11482512 TI - An unusual exostosis presenting as a bunion deformity. AB - A large exostosis was the source of a bunion deformity in a 60-year-old woman. Its unusual clinical and radiographic features were suggestive of a bizarre parosteal osteochondromatous proliferation. However, histologic features were most consistent with a benign osteocartilaginous exostosis. PMID- 11482513 TI - Prevertebral abscess with extension into the hip joint. AB - We present a case of a paravertebral abscess with extension into the hip joint. The presumed conduit of infection in this case is the iliopsoas bursa, which has been shown to provide direct communication between the retroperitoneal space and the hip joint in up to 15% of cadaveric specimens. The patient was treated with emergent open irrigation and debridement of the hip joint, as well as percutaneous drainage of the prevertebral abscess. The patient remains minimally symptomatic at the 2-year follow-up. PMID- 11482514 TI - Surgical treatment of olecranon fractures. AB - Fractures of the olecranon are common injuries with no single accepted treatment. Numerous clinical factors and biomechanical studies guide the surgeon in determining a treatment plan. The goals of surgical management include anatomical reduction and stable fixation, which allow early range of motion of the elbow joint. Numerous fixation methods have been described and include screw fixation, cerclage wiring, modified tension-band wiring, and plate fixation. The surgical technique depends on a combination of patient factors, the fracture pattern, and the mechanical stability of the fixation construct. Postoperative rehabilitation includes early active range-of-motion exercises and follows clinical and radiographic healing. Complications are related to the fracture, choice of implant, and surgical technique. PMID- 11482515 TI - Lateral plate-washer technique for revision tibiocalcaneal fusion. AB - Tibiocalcaneal fusion using intramedullary rods for severe end-stage Charcot deformity is challenging. Complications such as nonunion, delayed union, recurrent deformity, wound problems, and infection are common. Revision cases present the additional obstacle of obtaining bony purchase in the calcaneal screws. We present a technique using readily available instrumentation to obtain reliable calcaneal fixation. PMID- 11482516 TI - Assessing the reliability of PBPK models using data from methyl chloride-exposed, non-conjugating human subjects. AB - Physiologically based pharmacokinetic (PBPK) models are often optimized by adjusting metabolic parameters so as to fit experimental toxicokinetic data. The estimates of the metabolic parameters are then conditional on the assumed values for all other parameters. Meanwhile, the reliability of other parameters, or the structural model, is usually not questioned. Inhalation exposures with human volunteers in our laboratory show that non-conjugators lack metabolic capacity for methyl chloride entirely, and that elimination in these subjects takes place via exhalation only. Therefore, data from these methyl chloride exposures provide an excellent opportunity to assess the general reliability of standard inhalation PBPK models for humans. A hierarchical population PBPK model for methyl chloride was developed. The model was fit to the experimental data in a Bayesian framework using Markov chain Monte Carlo (MCMC) simulation. In a Bayesian analysis, it is possible to merge a priori knowledge of the physiological, anatomical and physicochemical parameters with the information embedded in the experimental toxicokinetic data obtained in vivo. The resulting estimates are both statistically and physiologically plausible. Model deviations suggest that a pulmonary sub-compartment may be needed in order to describe the inhalation and exhalation of volatile adequately. The results also indicate that there may be significant intra-individual variability in the model parameters. To our knowledge, this is the first time that the toxicokinetics of a non-metabolized chemical is used to assess population PBPK parameters. This approach holds promise for more elaborate experiments in order to assess the reliability of PBPK models in general. PMID- 11482517 TI - Effects of polybrominated diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) on thyroid hormone and vitamin A levels in rats and mice. AB - The ability of the commercial polybrominated diphenyl ether (PBDE) preparation Bromkal 70-5 DE to alter thyroid hormone and vitamin A levels as well as microsomal enzyme activities was compared with that of the commercial polychlorinated biphenyl (PCB) preparation Aroclor 1254 in orally exposed female rats (Sprague-Dawley) and mice (C57BL/6 N). Additional mice were exposed to the PBDE congener 2,2',4,4'-tetrabromodiphenyl ether (DE-47), or to the PCB congener 2,3,3',4,4'-pentachlorobiphenyl (CB-105). For 14 days the animals were given approximately isomolar daily oral doses of Aroclor 1254, CB-105 (both 10 mg/kg body weight), Bromkal 70-5 DE or DE-47 (both at 18 mg/kg body weight). In addition, further groups of rats and mice received a higher dose of Bromkal 70-5 DE, 36 mg/kg body weight. Bromkal 70-5 DE and DE-47 decreased plasma free and total thyroxine (T4) levels in both rats and mice, although with lower potency than that of Aroclor 1254 and CB-105. By contrast, thyroid-stimulating hormone (TSH) levels were not significantly changed in any of the groups. Reduction of hepatic vitamin A levels was seen in rats after Aroclor 1254 and Bromkal 70-5 DE exposure. A similar tendency was seen also in mice, but the effects were significant only for concentration data and not the total amount. Induction ofmicrosomal phase I enzymes, measured as ethoxy, methoxy and pentoxy resorufin O dealkylase (EROD, MROD, PROD) activities, was greatest after exposure to Aroclor 1254/CB-105 but were also significant in the Bromkal 70-5 DE/DE-47-treated groups. However, induction of uridine diphosphoglucuronosyl transferase (UDPGT) was small and for most groups insignificant. In conclusion, the PBDE compounds studied, although having a lower potency than the PCB compounds, decreased thyroxine and vitamin A levels and induced microsomal enzyme activities. Rats were more sensitive to the observed effects than mice. Microsomal phase I activity might be related, directly or indirectly, to the T4 and vitamin A effects, whereas several factors (such as weak enzyme induction and lack of correlation with altered T4 and vitamin A levels) argue against any UDPGT-related effects. PMID- 11482518 TI - Uptake of the dopaminergic neurotoxin, norsalsolinol, into PC12 cells via dopamine transporter. AB - The uptake of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, was studied in PC12 cells. The compound was actively taken up by the PC12 cells, with a Km value of 176.24 +/-9.1 microM and a maximum velocity of 55.6 +/- 7.0 pmol/min per mg protein; norsalsolinol uptake was dependent on the presence of extracellular Na+. The uptake of norsalsolinol was sensitive to two dopamine transporter inhibitors, GBR-12909 and reserpine, but was less sensitive to desipramine, a noradrenaline transporter inhibitor. Dopamine competitively inhibited norsalsolinol uptake into PC12 cells with a Ki value of 271.2 +/- 61.6 microM. These results suggest that norsalsolinol is taken up into PC12 cells mainly by the dopamine transporter. PMID- 11482519 TI - Effect of the neuroprotective agent riluzole on intracellular Ca2+ levels in IMR32 neuroblastoma cells. AB - Riluzole is an effective neuroprotective drug. Its effect on intracellular free Ca2+ levels ([Ca2+]i) has not been explored. This study examined the effect of riluzole on [Ca2+]i in IMR32 neuroblastoma cells using fura-2 as a Ca2+ probe. Riluzole 0.1-1 mM increased [Ca2+]i in a concentration-dependent manner. Removal of extracellular Ca2+ inhibited the response by 52 +/- 5%. The [Ca2+]i increase induced by 0.2 mM riluzole was unaltered by 0.1 mM La3+ or 10 microM verapamil, but was inhibited by 51 +/- 4% by 10 microM nifedipine. In Ca2+-free medium, pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) reduced the 0.2 mM riluzole-induced Ca2+ release by 44 +/- 3%; this reduction was augmented to 66 +/- 5% by additionally depleting the Ca2+ stores in the Golgi complex with 50 microM brefeldin A. Inhibition of inositol 1,4,5 trisphosphate formation by 2 microM U73122, a phospholipase C inhibitor, did not affect Ca2+ release induced by 0.2 microM riluzole. It was concluded that the neuroprotective agent riluzole increased [Ca2+]i in IMR32 neuroblastoma cells concentration-dependently by releasing Ca2+ from multiple stores in an inositol 1,4,5-trisphosphate-independent manner and also by inducing nifedipine-sensitive Ca2+ influx. PMID- 11482520 TI - Role of biological antioxidants in benzanthrone toxicity. AB - Previous studies indicate that benzanthrone, an anthraquinone dye intermediate, caused significant depletion of ascorbic acid (AsA). In this investigation the effect of benzanthrone on the status of different forms of AsA and other bio antioxidants such as glutathione (GSH) was studied. Oral administration of benzanthrone (50, 125 or 250 mg/kg body weight) resulted in a significant increase of urinary AsA levels with a concomitant decrease in the urinary dehydroascorbic acid (DHA) content in both rats and guinea-pigs. Benzanthrone caused a dose-dependent decrease in hepatic, adrenal and serum AsA levels with a subsequent increase in DHA and diketogulonic acid (DKA) levels in both rats and guinea-pigs. Following benzanthrone treatment, rats showed an increase in the scorbutic index (to 1.01-1.21) of the liver, adrenal glands and serum compared to controls (0.12-0.24). The scorbutic indices of liver, adrenal glands and serum were also substantially increased (to 3.61-11.20) in benzanthrone-treated guinea pigs compared to controls (0.16-0.38). Single oral administration of benzanthrone to guinea-pigs caused a dose-dependent depletion of GSH in liver (15-51%), adrenal glands (27-64%) and serum (32-86%). Furthermore, the depletion of GSH by benzanthrone in rats was of a lesser degree. This suggests that continued exposure of guinea-pigs to benzanthrone may lead to scurvy-type changes in this animal species but not to the same extent in rats, since the latter has the enzymatic capacity to synthesise AsA. Therefore, it can be hypothesised that benzanthrone per se, or its metabolites, interact with reduced GSH thereby causing its depletion. Furthermore, in order to replenish the depleted GSH levels, AsA might be oxidized to DHA and hence the decrease in AsA with the simultaneous increase in DHA was observed. PMID- 11482521 TI - Differential response of mast cells separated from various organs and basophils of dogs to the fluoroquinolone antimicrobial levofloxacin. AB - Histamine releases induced by the fluoroquinolone antimicrobial levofloxacin (LVFX) were investigated using mast cells separated from various organs and peripheral basophils of dogs, being the most susceptible species to quinolone derivatives, in both in vivo and in vitro systems. An intravenous infusion of LVFX at 30 mg/kg over a 30-min period produced endogenous histamine release from 5 min, and a maximum at 30 min, in which the plasma LVFX concentration was approximately 50 microM. A close correlation (r = 0.87, n = 20) between histamine and LVFX concentrations in plasma during the infusion was observed. In the in vitro study, LVFX at 30 microM or more caused histamine release from mast cells separated from the liver and skin, but not from the gastric mucosa, lung, and peripheral basophils. More exactly, the liver mast cells were most susceptible to LVFX among the organs tested. On the other hand, compound 48/80, a prototype histamine liberator, elicited the histamine release from the liver or skin mast cells at 10 microg/ml, and the calcium ionophore A23187 at 1 microM exhibited the histamine release from the mast cells derived from all organs examined. Histochemical analysis revealed that the liver and skin mast cells had positive reaction for both alcian blue and safranin staining, but the gastric mucosa and lung mast cells were only positive for alcian blue staining, indicating that LVFX preferably activated the connective tissue-type mast cells rather than the mucosal-type mast cells. The degranulation of the liver and skin mast cells brought about by either LVFX or compound 48/80, unlike the calcium ionophore A23187, was blocked by pretreatment with pertussis toxin, suggesting the involvement of pertussis toxin-sensitive G proteins. The results obtained from the canine experiments strongly suggest that LVFX induces histamine release from the connective tissue-type mast cells distributed mainly in the liver, somewhat in the cutaneous tissue, through the activation of pertussis toxin-sensitive G proteins. PMID- 11482522 TI - Bone marrow micronucleus assay in Brown-Norway rats exposed to diphenyl-methane 4,4'-diisocyanate. AB - Four groups of young adult male Brown-Norway rats (strain: BN/RijHsd) were either exposed whole-body (WB) to filtered air (negative control) or to respirable aerosols of monomeric diphenylmethane-4,4'-diisocyanate (MDI) at actual breathing zone concentrations of 9.2 +/- 1.5 and 118 +/- 8.6 mg/m3. One additional group was exposed to 11,0 +/- 14.4 mg/m3 MDI using a nose-only (NO) mode. Exposure was 1 h/day, one exposure per week on 3 consecutive weeks. MDI aerosols were generated using either a condensation (WB) or a dispersion-condensation (NO) principle with resultant MMADs of 2.4-3.1 microm and 1.2 microm (GSD approximately l.5), respectively. Humidity ranged from approximately 40% (WB) to approximately 5% (NO). Positive controls received cyclophosphamide and colcemid. Micronuclei in polychromatic erythrocytes (MN-PCE) were counted in bone marrow smears prepared after the final exposure on post-exposure days 1, 2 and 7 and stained with acridine orange or Wright-Giemsa. Both the WB-exposure regimen and the 7-day sampling time point were based upon a previous study in which a significant increase in MN-PCE was reported to occur. Rats exposed to 118 (WB) and 110 mg/m3 MDI (NO) exhibited signs of respiratory distress, including hypothermia, and increased lung weights when compared to WB-exposed rats. The intensity of changes appeared to be slightly more pronounced in NO-exposed rats. At no time point did this study provide any evidence of an MDI-induced effect on the frequency of MN-PCE. No differences in outcome existed following staining with acridine orange or Wright-Giemsa. There was an absence of any effect on the frequency of mast cells and their frequency was low enough not to interfere with the outcome of study. Positive control groups exhibited significant increases in MN-PCE. In summary, monomeric MDI aerosol did not induce cytogenetic damage in Brown-Norway rats when investigated according to current testing guidelines. PMID- 11482523 TI - Assessment of DNA damage in C6 glioma cells after antidepressant treatment using an alkaline comet assay. AB - The effects of acute (24 h) exposure to the antidepressants amitriptyline, imipramine (both tricyclics), fluoxetine (a selective serotonin re-uptake inhibitor) and tranylcypromine (a monoamine oxidase inhibitor) on DNA damage in cultured C6 rat glioma cells were determined using an alkaline comet assay. The effects of manipulation of intracellular cyclic AMP by pretreatment with dibutyryl cyclic AMP (dBcAMP) and 3-isobutyl-1-methylxanthine (IBMX) were also studied. For fluoxetine, the effects of addition of exogenous glutathione (GSH) and pretreatment with L-buthionine sulfoximine (BSO) were also assessed. There were increases in DNA damage with increasing concentrations of antidepressants. IBMX pretreatment protected against antidepressant-induced DNA damage in C6 cells pretreated with dBcAMP. Addition of exogenous reduced GSH and BSO increased DNA damage after fluoxetine exposure. The data show that the antidepressants induce significant amounts DNA damage in C6 cells. PMID- 11482524 TI - Kashin-Beck disease. PMID- 11482525 TI - Kashin-Beck disease: a historical overview. AB - In 1919, the first report on Kashin-Beck disease (KBD) made by a Japanese doctor described an endemic occurrence in the northern district of Korea. In the 1930s, Dr. Takamori and his colleagues at Manchuria Medical College produced a series of reports on its endemiology, clinical and roentgenological findings observed in the north-eastern district of China. In 1940s, a Tokyo University group led by Dr. Ogata found that the salivary glands of KBD patients were markedly degenerated. Administration of the condensed water taken from the endemic area into experimental rats produced degeneration of the salivary glands and changes in bones and joints similar to those of KBD. Thus, they proposed parotin deficiency theory as the etiology of KBD, and they recommended parotin therapy for KBD in its early stage. In the 1950s, Dr. Takizawa and his colleagues at Chiba University demonstrated that ferulic acid and p-hydroxy-cinnamic acid, found in the drinking water in the endemic area, caused degeneration of the salivary gland in rats. They recommended boiling the drinking water or using activated charcoal for the prevention of KBD. In the 1970s, the Japanese Ministry of Health and Welfare made a nationwide survey for the incidence of KBD in Japan. They concluded that there was no case of KBD in Japan with the exception of a few patients who had been brought up in the northeastern district of China and later had moved to Japan. PMID- 11482526 TI - Anthropometry and clinical features of Kashin-Beck disease in central Tibet. AB - We compared two different populations living in central Tibet with the purpose of establishing standard values for different anthropometric parameters in a rural population. Later on, these values were used as references for a similar study on a KBD population. One group (KBD) (n=1,246) came from the endemic areas, and the other group, serving as the control population (n=815), came from non-endemic areas. Both groups included children and adults and were of the Mongoloid type; they were farmers or semi-nomads. Height, weight, segment length, joint perimeter, joint diameter, joint movement were recorded. Also more subjective information such as general feeling of tiredness, rapid fatigue at work, work limitation, joint pain, muscle weakness, muscular atrophy, dwarfism, flatfoot, and waddling gate was also collected. Those variables were compared between the two groups. PMID- 11482527 TI - The anatomical distribution of radiological abnormalities in Kashin-Beck disease in Tibet. AB - A radiological study of osteoarticular changes in Kashin-Beck Disease (KBD) was undertaken on the appendicular skeleton in 105 patients with KBD, in 31 healthy subjects living in an endemic area and in 30 healthy subjects living in a non endemic area. The bone age was delayed in all three populations with no significant difference between the three studied Tibetan populations. Radiological changes occur in 56% of patients with KBD, and are usually bilateral. An analysis of the distribution of lesions shows a proximo-distal gradient. The changes are more common in the distal aspect of the limb and the lower limb is involved more commonly than the upper limb. The foot and ankle are involved in 89.5% of cases. The radiological changes and their distribution might be explained by the hypothesis of inhibition of angiogenesis by mycotoxins, exacerbated by chemical and physical environmental factors. PMID- 11482528 TI - Diagnostic, clinical and radiological characteristics of Kashin-Beck disease in Shaanxi Province, PR China. AB - Twenty clinical symptoms and four radiological signs of different joints in patients with Kashin-Beck disease (KBD) were studied in 2,560 subjects from endemic and non-endemic areas of the Shaanxi Province in China. It is suggested to classify the symptoms into five groups representing different manifestations of the disease. The association of some of the symptoms appears to provide significant criteria for use in the diagnosis of KBD. PMID- 11482529 TI - Histology of Kashin-Beck lesions. AB - There are few papers in existence describing the histopathology of Kashin-Beck disease. The few existing papers mention chondronecrosis within the epiphyseal primodium and metaphyseal cartilage. In the present study, two series of samples were available for histology: supernumerary fingers removed from young subjects and intra-articular bodies collected in more advanced cases of the disease. The prevailing characteristic of the samples is the absence of vascularisation within the proximal cartilage end plate of the phalanx associated with an alteration of the epiphyseal bone formation. These observations suggest that Kashin-Beck disease could develop from an alteration of the angiogenesis of the metaphyseal cartilage resulting in degeneration with consequent joint dysplasia, which may be associated with a decrease in growth of the diaphyseal bones. PMID- 11482530 TI - A 4-year study of the mycological aspects of Kashin-Beck disease in Tibet. AB - In order to clarify the association between mycotoxin-producing fungi in food and Kashin-Beck disease (KBD), we examined the occurrence and contamination levels of fungi in samples of barley grain, from KBD-affected families and from unaffected families in endemic areas. A control area without the occurrence of KBD served as reference. The first results obtained in 1995 showed that total mesophilic fungal contamination of barley grain was consistently higher in families with KBD. Trichothecium roseum (Pers) Link ex gray, Dreschlera Ito and Alternaria Nees ex Fr. were the three most common fungi significantly associated with KBD. In 1996 we again observed a significant difference between affected and non-affected families, especially with Trichothecium roseum and Ulocladium Preuss. On this basis, measures to prevent KBD were suggested and a preventive program has been set up since 1998 in 20 new villages. PMID- 11482531 TI - The prevalence of mycotoxins in Kashin-Beck disease. AB - Mycotoxins are naturally occurring toxic chemical compounds produced by fungi infesting agricultural crops both during their growth and storage. Such secondary metabolites, when ingested, can produce toxic syndromes in humans. As it has been suggested that mycotoxins might be involved in the development of Kashin-Beck disease (KBD), we undertook a survey of barley grains of KBD-affected families and non-affected families in that country. We found, by thin layer chromatography, a hitherto unknown metabolite of Alternaria sp. This was especially common on the barley grains of KBD-affected families. PMID- 11482532 TI - Measurement of the bioactivity of interleukin and tumour necrosis factor in synovial fluid of Kashin-Beck disease. AB - We studied the bioactivity of interleukin (IL-1) and tumour necrosis factor (TNF) in the synovial fluid of 12 patients with Kashin-Beck disease (KBD), 8 patients with osteoarthritis (OA) and 8 normal controls. C57 female mouse T cell proliferation method was used to test the bioactivity of IL-1, and lethality method with L929 cells to test the bioactivity of TNF. The bioactivities of IL-1 and TNF in the synovial fluid from patients with KBD were higher than those in the synovial fluid of osteoarthritis and normal controls, indicating the participation of IL-1 and TNF in the pathogenesis of KBD. PMID- 11482533 TI - Kashin-Beck disease and iodine deficiency in Tibet. AB - We evaluated iodine and selenium status in 575 children between 5 and 15 years with Kashin-Beck disease from endemic and non-endemic areas. Of these 267 (46%) children had goiter. The proportion of subjects with goiter was higher in the villages with Kashin-Beck disease than in the control village. In the villages with Kashin-Beck disease, 105 (23%) of the subjects had a serum thyrotropin greater than 10 mU/l as compared with 3 (4%) in the control village. The percentages of low serum thyroxine values and low serum tri-iodothyronine were greater in the villages where Kashin-Beck disease was endemic than in the control village. The percentages of low urinary iodine concentration were significantly greater in the subjects with Kashin-Beck disease. The results suggest that in areas where severe selenium deficiency is endemic, iodine deficiency is a risk factor for Kashin-Beck disease. PMID- 11482534 TI - Kashin-Beck disease and drinking water in Central Tibet. AB - A cross-sectional survey was carried out in order to study the relationship between Kashin-Beck disease and drinking water. The average volume of the water containers was larger in families unaffected by the disease. Organic material was measured by ultraviolet (UV) spectroscopy. The UV absorbency was significantly lower in drinking water of unaffected families. Thus, the organic material in drinking water may play a role in the pathogenesis of Kashin-Beck disease. PMID- 11482535 TI - Ethno-agricultural approach to the rural environment in the prevention of Kashin Beck disease. AB - Kashin-Beck disease occurs in several villages of Tibet; however, its local importance varies greatly. The ecoclimatological as well as the phytogeographical framework of the studied area are presented. An ecological approach based upon the ethno-ecosystem concept was carried out in the vicinity of each village. This study identifies 18 vegetation units on a structural basis; they were named for the dominant plants of each unit. Half of them belonged to the aquatic milieu. The different factors controlling their distribution were also identified. Particular attention will be paid to the links between man and the environment, particularly regarding alternative food intakes and water supply access. PMID- 11482536 TI - Kashin-Beck disease: from etiology to prevention or from prevention to etiology? AB - The aetiology of Kashin-Beck disease (KBD) remains elusive. Four factors have been convincingly associated with the disease: selenium deficiency, iodine deficiency, grain contamination with mycotoxin-producing fungi, and water pollution with organic material and fulvic acid. The most credible studies from a scientific standpoint, i.e. randomised placebo controlled trials and observational cohort studies have either not been conducted or did not provide unequivocal demonstration in favour of any of these hypotheses. Many studies such as case-control, cross-sectional, "before-after", and even more so, ecological studies have been conducted. They merely produced weak evidence and fail to support any single factor to the exclusion of the others. The most scientifically sound studies have included animal models, laboratory experiments and pathology studies; however, these have only provided indirect evidence. Although none of the competing theories prevails when they are compared using a predefined and standard set of causality criteria (temporality, strength of the association, biological gradient, experimental evidence, biological plausibility, coherence, specificity and analogy), none should be discounted. This is an indication that a comprehensive and unifying theory is most likely to be multifactorial. Because the ultimate goal of those who are compelled by the challenge of KBD is to prevent its occurrence, a perfect understanding of its mechanisms is not indispensable for action. Well-conducted randomised intervention should be the priority of researchers as well as public health professionals to demonstrate what works and what does not. PMID- 11482537 TI - Epidemiological support for a multifactorial aetiology of Kashin-Beck disease in Tibet. AB - We carried out a cross-sectional study in 12 rural villages in order to identify the risk factors for Kashin-Beck disease in Tibet. Children aged 5-15 years (n=575) were examined and their corresponding houses were visited. Samples were collected in order to study fungal contamination of stored grain and the organic matter content of drinking water. Multivariate analysis was performed using logistic regression and population attributable fractions were computed to estimate the impact of each factor. The following variables were independently associated with the disease: age, gender, low socio-economic status, indicators of a poorly diversified diet, iodine deficiency and small water container size (with higher organic matter levels in small containers). Selenium deficiency was severe in all study subjects. The degree of fungal contamination of barley grain was related to the highest percentage of cases (65%) in a sample of the study population. Higher urinary iodine levels were not associated with decreasing prevalence rates when Alternaria sp. was isolated. The data that we report supports the hypothesis that Kashin-Beck disease occurs as a consequence of oxidative damage to cartilage and bone cells when associated with decreased antioxidant defence. Another mechanism that may coexist is bone remodelling stimulated by thyroid hormones whose actions can be blocked by certain mycotoxins. PMID- 11482538 TI - Selenium, iodine and fungal contamination in Yulin District (People's Republic of China) endemic for Kashin-Beck disease. AB - We studied the status of selenium, iodine and fungal contamination in 353 school children (age 5-14 years) from four rural villages in the District of Yulin. In three villages Kashin-Beck disease (KBD) was endemic, whereas there were no cases of KBD in the fourth village. Clinical, biological and radiological examinations (right hand) were performed and KBD was established by X-ray diagnosis. The prevalence rate of KBD was 30.2%, 44.2% and 45.3% in the three endemic villages. Mean hair selenium and urine iodine concentrations were lower in affected than in unaffected children and fungal contamination in cereal grains stored in families with KBD was more elevated than in families without KBD. Low hair selenium concentration and presence of fungal cereal contamination were significantly associated with an increased risk of KBD, but low urine iodine was not. PMID- 11482539 TI - Effects of physical therapy on patients with Kashin-Beck disease in Tibet. AB - A clinical trial of physical therapy treatment for patients suffering from Kashin Beck disease (KBD) has been carried out in Tibet. One-hundred and thirty-five patients with Kashin-Beck disease were allocated to either physical therapy (72 patients) or prescription of multivitamins (63 patients). The patients were followed for 4 years. This study suggested a beneficial effect of physical treatment. PMID- 11482540 TI - Skeletal deformities induced by the intraperitoneal administration of deoxynivalenol (vomitoxin) in mice. AB - The contamination of drinking water by organic acids, selenium deficiency and the ingestion of fungal mycotoxins are the three main aetiological factors in the development of Kashin-Beck disease. An avian tibial chondrodysplasia induced by mycotoxins has been reported. Deoxynivalenol (DON) is one of many mycotoxins produced by the most common contaminating species of fungi. The pattern of skeletal malformations induced by its administration intraperitoneally to pregnant mice is reported. Costo-vertebral segmentation abnormalities were the main deformities observed. The chondrodysplasia previously described was not seen. PMID- 11482541 TI - The effect of T-2 toxin on IL-1beta and IL-6 secretion in human fetal chondrocytes. AB - The effects of T-2 toxin on IL-1beta and IL-6 secretion in human fetal chondrocytes in vitro were investigated. The evaluation is realised on primary monolayer culture of human fetal epiphyseal chondrocytes with or without PMA stimulation. The levels of supernatant IL-1beta and IL-6 were analyzed by ELISA. As compared with their respective controls, we observed a significant increase of IL-Ibeta and IL-6 in supernatants of chondrocytes cultivated for 24 h with T-2 at 8 ng/ml after PMA stimulation; in the absence of PMA, IL-Ibeta was increased alone after 48 h. The results demonstrated that T-2 toxin could superinduce IL 1beta and IL-6 secretion in chondrocytes. All these data suggested that superinduction of cytokines might be one of the key mechanisms of chondrocyte injuries by T-2 toxin. PMID- 11482542 TI - Comparison of the isometric cervical extension strength and a cross-sectional area of neck extensor muscles in college wrestlers and judo athletes. AB - Increasing neck muscle strength can play an important role in preventing neck injuries in contact sports. The purpose of this study was to examine the actual conditions of the isometric cervical extension strength (ICES) and the cross sectional area (CSA) of neck extensor muscles in male athletes participating in college wrestling and judo. The subjects comprised 18 wrestlers and 37 judo athletes from Nippon Sports Science University in Japan. The ICES was measured at eight angles (126 degrees, 108 degrees, 90 degrees, 72 degrees, 54 degrees, 36 degrees, 18 degrees, 0 degrees ). Transverse slices of 10 mm thickness were obtained at the position of each intervertebral disc between C2 and C3, C3 and C4, C4 and C5 and C5 and C6 using magnetic resonance imaging. The ICES of the wrestlers were significantly higher than those of the judo athletes. The ICES curve against the angle in wrestlers tended to differ from that of judo athletes. The CSA of neck extensor muscles in the wrestlers was significantly larger at all intervertebral levels examined than those of the judo athletes. A significant difference was observed in the CSA of the deepest area of neck extensor muscles between the groups although the difference was not significant in the superficial area. In this study, the ICES and the CSA in wrestlers were shown to be significantly higher and larger respectively than in the judo athletes, indicating a significant difference between these two sports. PMID- 11482543 TI - Fibre type proportion and fibre size in trapezius muscle biopsies from cleaners with and without myalgia and its correlation with ragged red fibres, cytochrome-c oxidase-negative fibres, biomechanical output, perception of fatigue, and surface electromyography during repetitive forward flexions. AB - In the literature enlarged/increased cross-sectional area (CSA) of type I muscle fibres has been reported as a morphological mark of work-related localised myalgia in the descending part of the trapezius muscle of women. These studies did not use enough subjects or lacked an adequate control group. The recording of surface electromyograms (EMG) is central to the research field of work-related myalgia. However, the influence of intrinsic muscle properties such as the effect of muscle fibre distribution upon surface EMG has to be better understood in order properly to evaluate this method as a possible diagnostic and preventive tool. This study had two aims. Firstly, it investigated the muscle fibre distribution and CSA in work-related myalgia in trapezius muscles. Secondly, the multivariate relationships among muscle morphology and histochemistry [ragged-red (RR) fibres, and cytochrome-c-oxidase-negative-fibre changes] EMG, perceived fatigue, and biomechanical output of shoulder flexions were analysed. The raw data have been presented in an earlier study. The participants in this study were 25 female cleaners with work-related myalgia of the trapezius muscle and 25 female cleaners not experiencing work-related myalgia of this muscle. The control group comprised 21 healthy female teachers who had not been exposed to highly repetitive work or static muscle work. Smaller CSA of type II fibres were found in cleaners compared to teachers. In this study the CSA of type I fibres of the trapezius muscles associated with myalgia were no greater than in muscles without myalgia. The prevalence of RR fibres together with age, fibre type proportions, CSA and working as a cleaner correlated with the ability to relax as recorded electromyographically. The relative mean frequency of the EMG of the trapezius muscle correlated with the prevalence of RR fibres, but it did not correlate with the proportions and CSA of different fibre types. Low biomechanical outputs and low signal amplitude increases of the EMG during the test were associated with high proportions of type -IIB fibres. The smaller CSA of type II fibres in cleaners might have reflected a different muscle activation pattern due to different occupational demands in cleaners than in teachers. Morphological or histopathological variables can influence the three EMG variables investigated. PMID- 11482544 TI - Patterns of muscle activation in human hopping. AB - In the present study, we examined the electromyogram (EMG) patterns of the soleus and medial gastrocnemius (MG) muscles during rhythmical, two-legged hopping to investigate the contributions of the monosynaptic short- and long-latency stretch reflexes during such a natural movement in human. During rhythmical hopping, soleus muscle is activated reflexly at near-monosynaptic latency by stretch resulting from passive ankle flexion upon landing. Soleus muscle also contracts voluntarily in order to launch the body into the next hop. This is part of the rhythmical bursts of activity producing the hops. Depending on the hopping interval, this phase of activation can follow the short-latency phase or precede landing at very short hopping intervals. In MG, there is an initial phase of activity that stiffens the muscle in preparation for landing, and continues through the contact phase. The monosynaptic reflex response to landing is usually superimposed on this activity. Depending on the hopping interval, both of these responses may be overlaid with activity that is time-locked to the take-off into the next hop, and serves to launch the body into the next hop. However, no evidence for a long-latency stretch reflex was found. In addition, the preferred hopping frequency for all subjects was about 2 Hz. This frequency is associated with a pattern of EMG activity the timing of which indicates that it balances the requirement for a comfortable landing from a hop with the optimal muscle activation required for launching the following hop. PMID- 11482545 TI - Exercise-induced neuromuscular dysfunction under reflex conditions. AB - The purpose of this research was to describe further the effects of exercise induced muscle damage on reflex sensitivity. The subjects were eight physically active, but untrained males, between the ages of 18 and 29 years. The effects of eccentric and concentric exercise on patellar tendon reflex responses were determined. The 8 week experiment consisted of two, 5 day, test protocols with a 6 week wash-out period between test protocols. Each 5 day test protocol consisted of the following six test sessions: (1) day 1--baseline, (2) day 2 baseline, (3) day 2--immediate post-exercise, and (4-6) days 3-5: 24, 48, and 72 h post exercise. On day 2, the subjects made either 100 fatiguing concentric or eccentric isotonic contractions using the right leg at 75% of the corresponding repetition maximum values. During each test session, the electromyogram (EMG) and force-time characteristics of basic and conditioned patellar tendon reflex responses were measured. The reflex amplitudes of basic and conditioned patellar tendon reflex responses were decreased following fatiguing concentric exercise. There were no immediate effects of fatiguing eccentric exercise on the basic and conditioned patellar tendon reflex responses, but the EMG amplitudes of these reflex responses were reduced on the days following eccentric exercise. The amount of conditioned patellar tendon reflex facilitation was decreased following the concentric exercise protocol and at 48 h post-eccentric exercise. Our conditioned reflex data suggest that post-exercise changes to the physiological mechanisms that modulate the recruitment gain of the alpha-motoneuron pool may depend upon the type of fatiguing exercise. PMID- 11482546 TI - REM sleep and catecholamine excretion: a study in elite athletes. AB - We investigated the potential influence of catecholamines on sleep architecture in endurance-trained athletes. The hypothesis was that endogenous levels of aminergic neurotransmitters influence sleep architecture. Thirteen well-trained male street cyclists (all members of the German national amateur team, mean age 23.9 years, mean body mass index 21.9 kg/m2) completed the protocol. Each subject was studied during training after a race competition (C) and later in a recovery/rest period (R) with no training and no competition. Polysomnography (PSG) was performed for one night (C) and for a second night some weeks later (R). Urinary levels of catecholamines collected during the preceding day and over the night of PSG were used as an index of excretion rate of circulating adrenergic agonists. Nighttime and daytime excretion of epinephrine and norepinephrine was significantly elevated after exercise (C vs R; P<0.01). Rapid eye-movement sleep (REM) onset latency was significantly increased (P=0.03) and REM was significantly decreased in the first half of the night in the training compared to the resting condition (C vs R, P=0.05). REM latency was correlated with increased epinephrine excretion on the day of exercise (C, r=0.63, P=0.02). The temporal appearance of REM during the night appears to be affected in part by the intense exercise associated with race competition, and urinary catecholamines are markers that are correlated with this alteration in REM appearance. PMID- 11482547 TI - The cumulative heat strain index--a novel approach to assess the physiological strain induced by exercise-heat stress. AB - The cumulative heat strain index (CHSI) is a new approach for assessing the total physiological strain experienced by subjects exposed to an exercise-heat stress. The index is based on inherent physiological logic that combines the thermoregulatory strain, which is described by the area under the hyperthermic curve, and the circulatory strain, which is characterized by heart-beat count. According to this model, the index reflects the dynamics of changes in the thermoregulatory and cardiovascular components and accounts for the complementary nature of the interaction between them. Mathematically, the index is calculated as follows: CHSI =[ sigma(0-t) hb-fc(0) x t] x l0(-3) x [ integral (0-t) Tre x dt Tre(0) x t] (units) Where: hb=heart beats, fc(0)=initial lowest heart rate (bpm), Tre = rectal temperature (Tre(0) = baseline Tre) (degrees C) and t = time (min) from the onset of measurements. Four sets of data, from various former studies, have been used to demonstrate the index's applicability and its sensitivity to differentiate between levels of stain under various stressful conditions (e.g. clothing insulation, acclimation to heat and levels of tolerance to heat). In all cases, the index was found to be a sensitive tool for assessing the level of strain. Furthermore, the CHSI can be used to predict potential strain. The index's high sensitivity arises from its nature, which reflects miniature differences in the pattern of changes in the dynamics of physiological responses and therefore is a powerful and practical tool for evaluating even minor changes in strain. PMID- 11482548 TI - Effects of oral creatine supplementation on maximal pedalling performance in older adults. AB - This study was conducted to investigate the effects of giving short-term doses of creatine by mouth to healthy older male subjects, taking into account their training status. A group of 42 volunteers was divided into three: a sedentary group composed of elderly sedentary men [n = 14, mean age 70.1 (SEM 1.2) years], a trained group composed of elderly trained cyclists [n = 14, mean age 66.4 (SEM 1.4) years] and a young group composed of young sedentary men [n = 14, mean age 26.0 (SEM 1.2) years]. In each group, double-blind randomization was carried out: one half was given creatine (3x5 g x day(-1)), and the other was given an iso nitrogenated placebo (3x10 g x day(-1)). Before and after the 5 days during which the supplements were given, all subjects performed five all-out 10-s sprints separated by 60-s intervals of passive recovery, seated on a cycle ergometer. Power output, work done and heart rate data were recorded during each sprint. The elderly and the young sedentary subgroups given creatine showed significant (P < 0.05) improvements in maximal power (+3.7% and +2.0%, respectively) and work done (+4.1% and +5.1%, respectively) in the subsequent tests. In contrast, no significant change in pedalling performances was observed in the trained elderly subjects. The creatine did not change the exercise and recovery heart rate profiles, in any group. Our study suggested that creatine given by mouth increases the anaerobic power and work capacity of sedentary people of different ages during maximal pedalling tasks. However, the level of physical activity seems to be a determinant of the ergogenic effect of creatine in older subjects. PMID- 11482549 TI - Effects of resistance training in humans on neck muscle performance, and electromyogram power spectrum changes. AB - The purpose of this study was to quantify the neuromuscular cervical adaptations to an 8 week strength training programme. Seven healthy men, with no pathological conditions of the neck, performed a lateral flexion isometric resistance-training programme three times a week. The training sessions consisted of one set of ten contractions, each of 6 s duration, at 60% of the predetermined maximal voluntary isometric torque (MVTim) (warm-up) and two sets of eight contractions, each of 6 s duration, at 80% MVTim. The training effects were evaluated in three ways: muscle size, strength and fatigability. The cross-sectional areas (CSA) of the trapezius (TRP) and sternocleidomastoideus (SCM) muscles were determined using a computerised tomographic scanner. Results showed an increase in the CSA of TRP and SCM muscles after training, 8.8% at C5 level and 6.4% at C7 level for SCM muscle and 12.2% at C7 level for TRP muscle. Strength increased significantly under both isometric and isokinetic conditions (35% and 20%, respectively). Muscle fatigability in lateral flexion was quantified during a sustained isometric contraction at 50% of MVTim. The shift of the mean power frequency of the electromyogram power spectrum density function of SCM muscle toward lower frequencies was less after training (14.6% compared to 6.8%). These results indicate the beneficial effect of a strength-training programme which increases neck muscle size and strength during lateral flexion, and decreases the fatigability of the superficial muscles of the neck. PMID- 11482550 TI - Oxygenation trends in vastus lateralis muscle during incremental and intense anaerobic cycle exercise in young men and women. AB - The purposes of this study were to compare the acute cardiorespiratory responses and muscle oxygenation trends during incremental cycle exercise to exhaustion with those observed during 30 s and 45 s Wingate tests in healthy men and women, and to examine the relationships between selected variables among these tests. Seventeen healthy junior badminton players, nine men [mean age, height, body mass and maximal oxygen uptake (VO2max) were 15.8 (SD 0.8) years, 1.73 (SD 0.08) m, 65.6 (SD 6.3) kg and 50.6 (SD 6.9) ml x kg(-1) x min(-1) respectively] and eight women [mean age, height, body mass and VO2max were 16.6 (SD 1.0) years, 1.65 (SD 0.03) m, 62.7 (SD 4.5) kg and 42.0 (SD 5.0) ml x kg(-1) x min(-1) respectively] completed a stepwise incremental exercise test to voluntary exhaustion and two Wingate tests lasting 30 s and 45 s in three separate sessions in random order. Cardiorespiratory responses were monitored breath-by-breath using a metabolic cart interfaced with an electrocardiogram. Tissue absorbancy trends were continuously recorded from the right vastus lateralis muscle using dual wavelength near infrared spectroscopy. Oxygen uptake and heart rate were significantly higher during the incremental test when compared to the two Wingate tests in the men and women. However, the oxygen pulse (oxygen utilization per heart beat, i.e., the product of stroke volume and arterio-venous oxygen difference) was not significantly different among the three tests in both sexes. The minimal tissue absorbancy, an index of muscle deoxygenation, was also not significantly different among the three tests in both sexes. Significant relationships were observed for the oxygen uptake (r2=0.72) and oxygen pulse (r2=0.60) between the incremental and 45 s Wingate tests in the sample for both sexes combined. The minimal tissue absorbancy, however, was not significantly related between the two tests. It was concluded that the significantly higher oxygen uptake during the incremental test was due to the higher heart rate because: firstly, oxygen pulse was not significantly different among the three tests, and secondly, peripheral factors, as indicated by the changes in muscle oxygenation, were not significantly different among the three test conditions. Although the peak values of the oxygen pulse during the incremental and 45 s Wingate tests were significantly correlated, the common variance of the minimal tissue absorbancy measurements between these two tests was quite low, suggesting considerable variation in the peripheral contribution during these two tests. PMID- 11482551 TI - Bilateral deficit during short-term, high-intensity cycle ergometry in girls and boys. AB - Peak torque during an isokinetic bilateral knee extension or flexion is usually lower than the summed peak torque of two unilateral contractions, and this phenomenon is designated as bilateral deficit (BD). The objectives of this study were to determine whether a BD also exists for more complex, alternating movements (e.g., short-term, high-intensity cycling), and to assess the possible effects of gender and maturity. Forty children and adolescents performed two unilateral (ULWAnT) and one bilateral Wingate anaerobic test (BLWAnT). The highest power reached in any 3-s interval (peak power; PP), and the total mechanical work (TMW) performed during the test was determined. Individual BD for PP (BDpp, %) was calculated by dividing the PP in the BLWAnT by the sum of the PP values of both ULWAnTs. The BD for TMW (BDTMw) was calculated in the same way. The sum of PP generated during the two ULWAnTs was larger than the PP measured in the BLWAnT (P<0.000001). The same difference was observed for TMW (P<0.0001). Maturity did not affect the BDpp or BDTMW (P=0.34). However, the BD was more pronounced in females compared to males (P= 0.03). In conclusion, a BD is evident during cycling exercise. PMID- 11482552 TI - Effects of aerobatics flight on oxygen consumption and heart rate control: influence on autonomic cardiovascular regulation during recovery. AB - Oxygen consumption (VO2) and blood pressure regulation were measured on five pilots during and after normal training aerobatics flights of a mean duration of 35 min. The acceleration vector along the longitudinal axis of the body (Gz) ranged from + 6.5 Gz to -3.5 Gz. VO2 was continuously monitored by a miniature telemetric system (K2). Heart rate (fc), the abdominal muscle electromyogram (EMG) and Gz levels were recorded synchronously on a magnetic tape recorder. A tilt test was performed pre- and post-flight to evaluate fc and blood-pressure variability. The left forearm blood flow was measured by strain-gauge plethysmography. The mean VO2 during flight was 1.2 l x min(-1), with a peak VO2 of 2.1 l x min x fc ranged between 55 and 165 beats x min(-1) and showed a progressive increase under the effect of + Gz, with a sudden fall during -Gz. The abdominal muscle EMG indicated the occurrence of muscle contraction under Gz load. Maximal responses were observed during the Gz phase. Comparison between pre and post-flight data showed lower post-flight systolic blood pressure with higher fc. Before flight, upright tilt induced a significant increase in low/ high frequency fc, as assessed using spectral analysis. This change was suppressed after flight. In summary, these data show that aerobatics flight leads to enhanced energy expenditure, mainly because of increased skeletal muscle work. The post-flight tilt test showed that aerobatic flight favors parasympathetic drive and, consequently, modifies blood pressure regulation during recovery. This action may decrease + Gz tolerance to a second aerobatics flight performed shortly after the first. PMID- 11482553 TI - Short-term changes in the series elastic component after an acute eccentric exercise of the elbow flexors. AB - We have studied the effect of a unique eccentric exercise session on the series elastic component (SEC) properties of human elbow flexors. Ten active females performed five sets of ten maximal eccentric contractions on an isokinetic ergometer. Maximal isometric (MVC) torque, and the corresponding myoelectrical activity (RMS) of the biceps brachii and the triceps brachii muscles were recorded before, immediately after, 48 h after, and 1 week after the exercise session. SEC compliance was also measured, using the in situ quick-release technique. Maximal eccentric torque declined significantly among the five sets of the exercise session (P < 0.01). The exercise session induced a significant decrease of the MVC over the post-exercise 48-h period. MVC returned to its control value 1 week after the exercise session. The neuromuscular efficiency calculated from the MVC torque:RMS ratio did not change throughout the experimental period, suggesting an optimisation of the neural drive to muscle force production capacity. The compliance values increased when force values decreased, but the slope of the relationship between both compliance and force logarithmic values was not significantly modified by the experimental procedure, indicating that the intrinsic properties of the SEC were not affected by the eccentric exercise session. The contractility-elasticity coupling was also maintained at a constant level during the recovery period. In other words, after an acute eccentric exercise, the evolution of SEC compliance was closely linked to force changes. PMID- 11482554 TI - Magnitude and decay of stretch-induced enhancement of power output. AB - There is ample evidence that the pre-stretching of a muscle enhances performance of the subsequent shortening or concentric contraction. The purpose of this study was to investigate this stretch-induced enhancement in terms of the effect of load on the magnitude and decay of the muscle's power output. Instantaneous, mean and peak power outputs were compared between concentric-only and rebound bench press throws at different loads. The maximal strength [1 RM = 84.0 (10.5) kg] of 18 male subjects [21.1 (3.0) years, 87.2 (11.9) kg] was determined using an isoinertial Smith press machine. The results of this study indicate enhancement of concentric motion by prior eccentric muscle action (200-780% enhancement in the first 100 ms). However, the magnitude and temporal characteristics of this enhancement differed across loads. Using a lighter load produced greater initial enhancement (P < 0.05) but the decay of this enhancement was more rapid. The heavier 80% 1 RM loading showed a later time to peak enhancement (80 ms versus 20 ms) and slower decay of the stretch induced augmentation (460 ms versus 260 ms). It is concluded that the degree of stretch-shorten cycle enhancement differs according to the load and the time-dependent characteristics of the motion. PMID- 11482555 TI - Respiratory muscle endurance training in humans increases cycling endurance without affecting blood gas concentrations. AB - Isolated respiratory muscle endurance training (RMT) can prolong constant intensity cycling performance. We tested whether RMT affects O2 supply during exercise, i.e. whether the partial pressure of oxygen in arterial blood (Pa,O2) and/or its oxygen saturation (SaO2) are higher during exercise after RMT than before. A group of 28 sedentary subjects were randomly assigned to either an RMT (n = 13) or a control group (n = 15). The RMT consisted of 40x30 min sessions of normocapnic hyperpnoea. The control group did not perform any training. Breathing and cycling endurance time as well as PaO2 and SaO2 during cycling at a constant intensity of 70% maximum power output were measured before and after the RMT or the control period. Mean breathing endurance increased significantly after RMT compared to control [RMT 5.2 (SD 2.9) vs 38.1 (SD 6.8) min, control 6.5 (SD 5.7) vs 6.4 (SD 7.6) min; P < 0.01], as did mean cycling endurance [RMT 35.6 (SD 11.9) vs 44.0 (SD 17.2) min, control 32.8 (SD 11.6) vs 31.4 (SD 14.4) min; P<0.05]. The RMT did not affect PaO2 which ranged from 11.6 to 12.3 kPa (87-92 mmHg), and SaO2 which ranged from 96% to 98% throughout all tests. In conclusion, RMT substantially increased breathing and cycling endurance in sedentary subjects. These changes, however, cannot be attributed to increased O2 supply, as neither PaO2 nor SaO2 were increased during exercise after RMT. PMID- 11482556 TI - Whole-body pre-cooling does not alter human muscle metabolism during sub-maximal exercise in the heat. AB - Muscle metabolism was investigated in seven men during two 35 min cycling trials at 60% peak oxygen uptake, at 35 degrees C and 50% relative humidity. On one occasion, exercise was preceded by whole-body cooling achieved by immersion in water during a reduction in temperature from 29 to 24 degrees C, and, for the other trial, by immersion in water at a thermoneutral temperature (control, 34.8 degrees C). Pre-cooling did not alter oxygen uptake during exercise (P > 0.05), whilst the change in cardiac frequency and body mass both tended to be lower following pre-cooling (0.05 < P < 0.10). When averaged over the exercise period, muscle and oesophageal temperatures after pre-cooling were reduced by 1.5 and 0.6 degrees C respectively, compared with control (P < 0.05). Pre-cooling had a limited effect on muscle metabolism, with no differences between the two conditions in muscle glycogen, triglyceride, adenosine triphosphate, creatine phosphate, creatine or lactate contents at rest, or following exercise. These data indicate that whole-body pre-cooling does not alter muscle metabolism during submaximal exercise in the heat. It is more likely that thermoregulatory and cardiovascular strain are reduced, through lower muscle and core temperatures. PMID- 11482557 TI - Implication of milk and milk products in food-borne diseases in France and in different industrialised countries. AB - A study was carried out to estimate the proportion of diseases due to milk and milk products among food-borne diseases recorded in France and in other countries since 1980. Particular attention was given to whether the milk involved was heat treated or not. Four etiologic agents were considered: Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, and pathogenic Escherichia coli. An overview of food-borne disease annual reports from seven countries indicated that milk and milk products were implicated in 1-5% of the total bacterial outbreaks; however, details about the type of product and milk involved were usually not provided. When considering 60 outbreaks and four single cases described in the literature and implicating milk and milk products, confirmed or suspected food vehicles were distributed as follows: milk, 39.1%, cheese, 53.1%, other milk products, 7.8%. Overall, 32.8% of the food vehicles were made from pasteurised milk; 37.5% from raw milk; 10.9% from milk stated as "unpasteurised"; and 18.8% from unspecified milk. Salmonella spp. were responsible for 29 outbreaks, L. monocytogenes for 10 outbreaks and four well-documented single cases, pathogenic E. coli for 11 outbreaks, and S. aureus for 10 outbreaks. Analysis of unpublished data about food-borne disease outbreaks, listeriosis excluded, collected by the coordinator of the French surveillance system from 1992 to 1997, revealed 69 documented outbreaks for which milk and milk products were confirmed as the vehicle by the isolation of the etiologic agent. The food vehicles were distributed as follows: milk, 10%; cheese, 87%; others, 3%. UHT milk accounted for 1.5%, raw milk and raw milk products for 48%, and milk and milk products from unspecified milk for 50.5% of the 69 outbreaks. S. aureus was by far the most frequent pathogen associated with these outbreaks (85.5% of the outbreaks), followed by Salmonella (10.1%). This study demonstrates the limitations of the surveillance systems and the difficulties in estimating the contribution of milk and milk products to food-borne diseases. In particular, it was not possible to find out in many outbreaks what heat treatment, if any, the milk had undergone. PMID- 11482558 TI - Evaluation of the thermal resistance of different Salmonella serotypes in pork meat containing curing additives. AB - The preliminary heat resistance evaluation of 94 Salmonella strains was carried out in culture medium (Trypticase soy broth, TSB). The heat resistance of three S. typhimurium strains (ATCC 14028, 133 and 1116), a strain each of S. derby B4373, S. potsdam 1133, S. menston 179. S. eppendorf 166, and S. kingston I124 was determined also in pork meat containing curing additives. As expected, the eight Salmonella strains showed greater heat resistance in pork meat than in TSB. At the lowest temperature (58 degrees C), the heat resistance increased 1.5-4 times, and it was most pronounced for the strains being most heat sensitive in TSB. S. potsdam 133 was the most resistant strain in pork meat, with D-values at 58 degrees C, 60 degrees C and 63 degrees C of 4.80, 1.57 and 0.30 min, respectively. The most sensitive strain turned out to be S. kingston 1124, with D values of 2.79. 0.92 and 0.24 min, at the same temperatures. According to collected data, the heating processes, as applied to cured pork meat, providing an internal temperature of 60 degrees C for 9-10 min or of 63 degrees C for 3-4 min can be expected to provide a > or = 7 D kill of Salmonella belonging to the serotypes studied. PMID- 11482559 TI - Utilization at high pH of starter cultures of lactobacilli for Spanish-style green olive fermentation. AB - Inoculation at alkaline pH (above 9) of lye-treated green olives with starter cultures of Lactobacillus pentosus CECT 5138 was studied. Despite an initial loss of viability in the order of 1-2 log cycles on average, depending mainly on time of application, cultures grew and initiated an accelerated fermentation process. Inoculation reduced the population of Enterobacteriaceae, and thereby potential spoilage, and produced a quicker acidification of brines and decrease of pH, when compared with control uninoculated batches. Results obtained throughout three consecutive seasons demonstrated that utilization at high pH of starter cultures of lactobacilli is feasible, provided that the inoculum size takes into account the initial low survival. PMID- 11482560 TI - Comparison of VIDAS enzyme-linked fluorescent immunoassay using Moore swab sampling and conventional culture method for Salmonella detection in bulk tank milk and in-line milk filters in California dairies. AB - A comparison of the VIDAS Salmonella (SLM) assay using the manufacturer's recommended sampling method or a Moore swab sampling method demonstrated that the Moore swab method detected a greater number of positive samples (83.0% vs. 67.92%). When results using a conventional culture technique were compared to the VIDAS SLM assay using Moore swab sampling, there was good agreement (97.5%). When the VIDAS SLM assay was used to test in-line milk filters and compared to results from the conventional culture method, there was also good correlation between test results (95.57%). Overall, the VIDAS SLM assay using a modified sampling method compared favorably to the conventional culture method and had the advantages of taking less time to obtain a negative or presumptive positive result, being less technically complicated and requiring less screening of non lactose fermenting colonies from negative samples. PMID- 11482561 TI - Measurements and predictions of growth for Listeria monocytogenes and Salmonella during fluctuating temperature II. Rapidly changing temperatures. AB - Growth of Listeria monocytogenes and Salmonella was examined during various rates of increase and decrease in temperature from and to the minimum for growth. Growth was little affected by even the most rapid changes and injury or lag was not observed. Subsequent investigations of growth during periods of rapid variation in temperature from and to temperatures below the growth minimum again had little effect and growth was satisfactorily predicted using the dynamic model of Baranyi and Roberts [Int. J. Food Microbiol. 23 (1994) 277] in conjunction with the data of Food Micromodel. PMID- 11482562 TI - Enterotoxin production and DNA fingerprinting in Staphylococcus aureus isolated from human and food samples. Relations between genetic types and enterotoxins. AB - A total of 224 Staphylococcus aureus strains from human carriers (110 strains) and manually handled foods (114 strains) collected in the Principality of Asturias, Spain over 1995-1999 were analysed for the production of enterotoxins (SEs) A, B, C, and D by a reversed passive latex agglutination test and by amplification of ent genes (A, B, C, D, E, and J) using PCR. Sixty-two strains were enterotoxigenic and a good relation between detection of SEs and their ent genes was found. No strain carried entE and all strains producing SED carried entD and entJ genes. Among the enterotoxigenic strains the percentages registered were 29, 8, 35, 18, 2, 2, and 6 for SEA, SEB, SEC, SEDJ, SEAC, SEADJ and SECDJ, respectively. DNA fingerprinting of 77 strains (the SE prototypes, 62 enterotoxigenic and 10 non-enterotoxigenic [NE]) was carried out by randomly amplified polymorphic DNA using two selected primers independently. Combining results from both primers, 10 genetic types were defined, which showed a different degree of relationship (similarity coefficient: 0.9-0.36) and were clustered into three lineages. One lineage clustered five genetic types and a wide diversity of strains, mainly SEA, SEB, SEDJ, and NE. Another lineage clustered only SEC, SECDJ and NE strains. These two lineages showed a low genetic relationship and appeared as endemic in healthy humans living in the Principality of Asturias. The third lineage included only the prototype strains for SEA and SEE. PMID- 11482563 TI - Antimicrobial susceptibility of starter culture bacteria used in Norwegian dairy products. AB - Commercial starter culture bacteria are widely used in the production of dairy products and could represent a potential source for spread of genes encoding resistance to antimicrobial agents. To learn more about the antimicrobial susceptibility of starter culture bacteria used in Norwegian dairy products, a total of 189 isolates of lactic acid bacteria were examined for susceptibility to ampicillin, penicillin G, cephalothin, vancomycin, bacitracin, gentamicin, streptomycin, erythromycin, tetracycline, chloramphenicol, quinupristin/dalfopristin, ciprofloxacin, trimethoprim and sulphadiazine using Etest for MIC determination. Most of the isolates (140) originated from 39 dairy products (yoghurt, sour cream, fermented milk and cheese), while 49 were isolated directly from nine commercial cultures. The bacteria belonged to the genera Lactobacillus, Lactococcus, Leuconostoc and Streptococcus. Only one of the 189 isolates was classified as resistant to an antimicrobial agent included in the study. This isolate, a lactobacillus, was classified as high level resistant to streptomycin. The remaining isolates were not classified as resistant to the antimicrobial agents included other than to those they are known to have a natural reduced susceptibility to. Thus, starter culture bacteria in Norwegian dairy products do not seem to represent a source for spread of genes encoding resistance to antimicrobial agents. PMID- 11482564 TI - Occurrence of Campylobacter jejuni in vegetables. AB - In order to understand the importance of vegetables in the transmission of thermophilic Campylobacter, 56 samples of different vegetables were screened. Out of these, 2 samples (1 spinach and 1 fenugreek) revealed the presence of Campylobacter jejuni biotype I. Both the isolates were enteropathogenic in rat ileal loop test. PMID- 11482565 TI - Inhibitory effects of eugenol and thymol on Penicillium citrinum strains in culture media and cheese. AB - In the present work we studied the antifungal effect of eugenol and thymol on the growth and production of citrinin from Penicillium citrinum (NRRL 2274 and NRRL 2269) in culture media and in different Spanish cheeses (Arzua-Ulloa, Cebreiro and San Simon). The rate of growth was assessed by measuring colony diameters and the production of citrinin was measured using a rapid semi-quantitative fluorometric technique confirmed by RP-HPLC. A stronger inhibitory effect of eugenol than thymol was evident. 200 microg/ml of eugenol in solid culture medium increased the lag time of growth up to 9 days, and decreased the rate of colony growth. In liquid medium, a complete inhibition of fungal growth was observed. By contrast, thymol in the liquid culture medium only affected the growth rate. In Arzua-Ulloa cheese, 200 microg/ml of eugenol fully inhibited fungal growth, while in Cebreiro cheese no effect was observed for this compound. Regarding the capacity to inhibit mycotoxin production 100 microg/ml eugenol delayed citrinin production until the sixth day, after which a limiting effect persisted. In Arzua Ulloa cheese, no citrinin was detected at a concentration of 150 microg/ml of eugenol, but citrinin was detected after 5 days in the case of thymol at the same concentration. In Cebreiro cheese, neither eugenol nor thymol prevented the production of citrinin at the concentrations applied. PMID- 11482566 TI - DNA-based, culture-independent strategies for evaluating microbial communities in food-associated ecosystems. AB - Culture-independent molecular techniques are now available to study microbial ecosystems. They are opening interesting perspectives to problems related to composition and population dynamics of microbial communities in various environmental niches (e.g., soil, water) and foods. In fermented food products, estimates of true microbial diversity is often difficult chiefly on account of the inability to cultivate most of the viable bacteria. The increasing knowledge of gene sequences and the concomitant development of new culture-independent molecular techniques are providing new and effective tools to compare the diversity of microbial communities and to monitor population dynamics in minimally disturbed samples. In this review, recent advances in these techniques are reported. Possible applications to food-associated microbial ecosystems are emphasised. PMID- 11482567 TI - Microbiological and biochemical characteristics of Canestrato Pugliese cheese made from raw milk, pasteurized milk or by heating the curd in hot whey. AB - Canestrato Pugliese cheeses were produced from raw ewes' milk (R and R(II) cheeses), pasteurized ewes' milk (P cheese) and by heating the curd in hot whey according to a traditional protocol (T cheese). R(II) differed from R cheese mainly by having been produced from raw milk with a higher number of somatic cells, 950.000 vs. 750.000 ml(-1), respectively. Compared to P and T cheeses, R and R(II) cheeses had a higher concentration (one or two orders of magnitude) of cheese-related bacteria such as adventitious mesophilic lactobacilli, enterococci and staphylococci. At the end of ripening, all cheeses contained less than 1.0 log cfu g(-1) of total and fecal coliforms, and Escherichia coli and Staphylococcus aureus were not detected. As shown by phenotypic identification and RAPD-PCR, R cheese contained the largest number of mesophilic lactobacilli species and the greatest diversity of strains within the Lactobacillus plantarum species. Primary proteolysis did not differ appreciably among the cheeses. On the contrary, both urea-PAGE and the RP-HPLC analyses of the water-soluble N fractions showed the more complex profiles in cheeses produced by raw milks. R and R(II) cheeses had the highest values of water-soluble N/total N (ca. 30%) and the highest concentration of total free amino acids (ca. 40 mg g(-1) which approached or exceeded those reported for Italian cheeses with very high level of proteolysis during ripening. The main differences between R-R(II) and P-T cheeses were the concentrations of aspartic acid, proline, alanine, isoleucine, histidine and lysine. The water-soluble extracts of R and R(II) cheeses contained levels of amino-, imino- and di-peptidase activities, which were about twice those found in P and T cheeses. Cheeses differed slightly in the concentration of total free fatty acids that ranged between 1673 and 1651 mg kg(-1) in R and R(II) cheeses, and 1397 and 1334 mg kg(-1) in P and T cheeses. Butyric, caproic, capric, palmitic, oleic and linoleic acids were found at the highest concentrations. PMID- 11482568 TI - Comparison of dichloran 18% glycerol (DG18) agar with general purpose mycological media for enumerating food spoilage yeasts. AB - Dichloran 18% glycerol (DG18) agar was originally developed to enumerate xerophilic foodborne moulds. However, some laboratories are using DG18 agar as a general medium to enumerate foodborne moulds and yeasts. A collaborative study, with the participation of seven laboratories, was undertaken to compare DG18 agar with dichloran rose bengal chloramphenicol (DRBC) agar, tryptone glucose yeast extract chloramphenicol (TGYC) agar, and plate count agar supplemented with chloramphenicol (PCAC) for enumerating 14 species of common food spoilage yeasts. Comparison of the mean values of populations of all yeasts recovered on each medium revealed no significant differences among DRBC agar, PCAC, and TGYC agar, while each of these media supported the development of significantly (P < or = 0.05) higher numbers of colonies than DG18 agar. However, differences were only 0.08 to 0.10 log10 cfu/ml, making the practical significance questionable. The overall coefficient of variation (CV) for within laboratory repeatability was 1.71%, while the CV for reproducibility of counts obtained among laboratories was 6.96%. Compared to DRBC agar, TGYC agar, and PCAC, yeast colonies were smaller on DG18 agar. Growth of Brettanomyces anomalus, Cryptococcus albidus, and Rhodotorula mucilaginosa was particularly retarded or inhibited on DG18 agar. Based on the performance of media in supporting colony development and ease of counting colonies, the use of DG18 agar as a general enumeration medium for foodborne yeasts cannot be recommended. PMID- 11482569 TI - Comparison of selective media for the recovery and enumeration of probiotic yeasts from animal feed. AB - Six selective media (acidified malt extract agar, CHROMagar Candida, dichloran rose bengal chloramphenicol, molybdate, oxytetracycline glucose yeast extract and Petrifilm Yeast and Mould agar) were examined for the recovery of three yeasts commonly used in animal feeds as probiotic additives (Candida pintolopesii, C. saitoana and Saccharomyces cerevisiae). The highest recovery was obtained on oxytetracycline glucose yeast extract agar, although this was susceptible to overgrowth by moulds. CHROMagar Candida also gave good recovery and species were easily discriminated by the differential colour formation of colonies. PMID- 11482570 TI - A temperature-type model for describing the relationship between fungal growth and water activity. AB - Growth of Penicillium chrysogenum, Aspergillus flavus, Cladosporium cladosporioides and Alternaria alternata at their respective optimum temperatures was studied in Potato Dextrose Agar (PDA) medium at different water activities (a(w)) adjusted with glycerol. The growth rate (mu) was expressed as the increase in colony radius per unit of time. This paper extends the model that showed the relationship between temperature and bacterial growth rate developed by Rosso et al. [J. Theor. Biol. 162 (1993) 447] to describe the influence of a(w) on fungal development. An excellent correlation between the experimental data and the model predictions was obtained, the regression coefficients (r2) were greater than 0.990, with the exception of that for A. flavus (r2 = 0.982). In addition, the use of such a model allows predictions of the cardinal water activities: a(wmin), a(wopt) and a(wmax). The estimation of the minimum water activity (a(wmin)) was in accordance with data literature for all the moulds considered here, but seemed to be slightly underestimated for P. chrysogenum and A. flavus when compared to our experimental values. The estimations of the optimal water activity (a(wopt)) and the optimal growth rate (muopt) were in excellent agreement to the experimental results for the four moulds. Through this example, it is suggested that the same approach for modelling can be used for various microorganisms (e.g. bacteria and moulds), and different environmental parameters (e.g. temperature and water activity). PMID- 11482571 TI - Detecting and genotyping Escherichia coli O157:H7 using multiplexed PCR and nucleic acid microarrays. AB - Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detecting and genotyping E. coli O157:H7. The array was composed of oligonucleotide probes (25 30 mer) complementary to four virulence loci (intimin, Shiga-like toxins I and II, and hemolysin A). Target DNA was amplified from whole cells or from purified DNA via single or multiplexed polymerase chain reaction (PCR), and PCR products were hybridized to the array without further modification or purification. The array was 32-fold more sensitive than gel electrophoresis and capable of detecting amplification products from < 1 cell equivalent of genomic DNA (1 fg). Immunomagnetic capture, PCR and a microarray were subsequently used to detect 55 CFU ml(-1) (E. coli O157:H7) from chicken rinsate without the aid of pre enrichment. Four isolates of E. coli O157:H7 and one isolate of O91:H2, for which genotypic data were available, were unambiguously genotyped with this array. Glass-based microarrays are relatively simple to construct and provide a rapid and sensitive means to detect multiplexed PCR products; the system is amenable to automation. PMID- 11482572 TI - Effects of carbon and nitrogen sources, sodium chloride and culture conditions on cytotoxin production by Salmonella choleraesuis. AB - The effects of carbon and nitrogen sources, incubation temperature, shaking speed, initial pH of culture broth as well as various concentration of NaCl on the production of cytotoxin by Salmonella choleraesuis SC-5 were evaluated in the present study. Results reveal that the optimal temperature, initial medium pH and shaking speed for cytotoxin production was 37 degrees C, pH 6.0-8.0 and 100 rpm, respectively. Tryptone was the best of the eight nitrogen sources tested for toxin production by S. choleraesuis. Among the nine carbon sources tested, S. choleraesuis produced a higher amount of cytotoxin in media containing glucose, fructose, galactose, sorbitol or mannitol as the carbon source. No toxin was detected in broths containing 4.0% or more sodium chloride in Tryptic soy broth (TSB). Cultures of S. choleraesuis in the medium containing 2.0% tryptone, 0.5% NaCl, 0.25% K2HPO4 and 0.25% of the best carbon source under the optimal conditions for 14 h resulted in the highest cytotoxin production. The Vero cell CD50 of S. choleraesuis lysate of cells grown under these optimal conditions was a titer of 589-758 per mg of lysate protein. PMID- 11482573 TI - The effects of freezing and thawing on the survival of Escherichia coli O157:H7 in apple juice. AB - Unpasteurized apple juice, adjusted to pH 3.6 to 7.0 was inoculated (10(7) CFU/ml) with single strains of E. coli O157:H7 to evaluate the effect of frozen storage on the viability of this organism. Samples were stored under frozen conditions (-20+/-2 degrees C) for up to 16 days. Cell populations were determined at regular intervals by plating onto tryptic soy agar with added pyruvate (TSAP) or onto sorbitol MacConkey agar (SMA). Populations in the neutralized juice remained unchanged during frozen storage. Populations in non neutralized juice decreased by 1-3 log10 CFU/ml depending on the strain tested and the pH of the juice. The greatest population decrease was observed with the first freeze/thaw cycle of frozen storage (24 h) and a slow decline in survival occurred thereafter. Injury was observed after 2 weeks of storage when juice pH was at or below pH 4.2. When samples were subjected to multiple freeze/thaw cycles, loss of viability and injury increased with each freeze/thaw cycle. PMID- 11482574 TI - Inhibition of the binding of enterotoxigenic Escherichia coli Pb176 to human intestinal epithelial cell line HCT-8 by an extracellular protein fraction containing BIF of Bifidobacterium longum SBT2928: suggestive evidence of blocking of the binding receptor gangliotetraosylceramide on the cell surface. AB - The extracellular protein fraction (P100V) containing the protein BIF produced by Bifidobacterium longum SBT2928 (BL2928), which inhibits the binding of enterotoxigenic Escherichia coli Pb176 (ETEC) to the glycolipid binding receptor gangliotetraosylceramide (GA1) also inhibited the binding of ETEC to the human intestinal epithelial cell line HCT-8 (ATCC CCL 244) in a dose-dependent manner. ETEC-binding inhibitory experiments using crude colonization factor antigen (CFA) II prepared from ETEC, rabbit anti-GA1 antiserum, medium containing GA1 and media containing lectins, as the binding-inhibitors, suggest that the interaction between the CFA-II antigen present on the cell surface of ETEC and GA1 expressed on HCT-8 cells plays a significant role in the adherence between them. It is strongly suggested that the P100V fraction works as a blocker for the ETEC receptor GA1 on HCT-8 cells. PMID- 11482575 TI - Benign spindle cell tumors of the mammary stroma: diagnostic criteria, classification, and histogenesis. AB - Purely benign mesenchymal spindle cell neoplasms of the breast are currently labeled under various terms in the literature (benign spindle cell tumor, fibroma, spindle cell lipoma, myofibroblastoma, solitary fibrous tumor, myogenic stromal tumor). The lack of strict diagnostic criteria to clearly indicate such mesenchymal neoplasms is the main reason which generated the risk of terming the same lesion under different names or, conversely, of collecting different types under the same term. Although such neoplasms exhibit morphological and immunophenotypical heterogeneity, they actually represent variations of the same tumor entity, likely arising from the uncommitted vimentin+/CD34+ fibroblasts of the mammary stroma, capable of multidirectional mesenchymal differentiation. To cover the entire spectrum of such lesions, the term "benign spindle cell tumors (BSCTs) of the mammary stroma" is advocated. BSCTs can be subtyped into four main groups by light microscopy (LM) and immunocytochemistry (ICC): fibroblastic, myofibroblastic, fibrohistiocytic, and mixed forms. A simple and practical approach to a nosologically correct diagnosis and a list of differential diagnoses are presented. The awareness of the diversity of morphological and immunophenotypical features of BSCTs of the mammary stroma, including uncommon variants, is helpful to avoid confusion with other monomorphic bland-looking benign and malignant spindle cell tumors and tumor-like lesions of the breast. PMID- 11482576 TI - Consequences of a National Mammography Screening Program on diagnostic procedures and tumor sizes in breast cancer. A retrospective study of 1540 cases diagnosed and histologically confirmed between 1995 and 1997. AB - In 1992, a national screening mammography program, including female patients between 50 and 64 years of age, was launched in Luxembourg. The effects of this campaign on the different diagnostic procedures, especially fine needle aspirations (FNA), large core needle biopsies (LCNB), and surgical specimens, were analyzed. From 1983 to 1997, the National Cancer Registry recorded 3167 new cases of invasive female breast cancer, all histologically diagnosed in one central pathology department. In 1996, the population consisted of 418,300 inhabitants (212,900 females). The number of breast cancer, tumor size, the nature of the diagnostic procedures, their diagnostic value as well as the number of physicians, "aspirators", and "biopsists" were evaluated. Between 1992 and 1994, the incidence of invasive breast cancers increased, concomitant with the launching of a National Screening Mammography Program. The diagnosis of in situ cancers tripled, and the mean size of invasive breast cancer decreased from 2.1 2.4 cm to 1.1-1.4 cm. Since 1994, the number of FNA had remained stable, LCNB had increased by 417.5%, and surgical biopsies had decreased by 18.95%. Between 1995 and 1997, 28.37% of 1075 FNA, and only 9.6% of 465 LCNB yielded inadequate samples. FNA were done by 77 different doctors (53.25% being gynecologists) and LCNB by 34 (52.94% being radiologists). The first diagnoses of all invasive cancers (n = 790) were made by using frozen sections from surgical specimens in 58.35% (n = 461), LCNB in 18.23% (n = 144), mastectomy in 10.13% (n = 80), formalin-fixed biopsies in 9.49% (n = 75), and FNA in 3.17% (n = 25). There are beneficial effects (increase in the number of diagnoses of in situ cancer; decrease in tumor sizes) not only for the "target" age group (50-64 years), but also for all female age groups (> 15 years). For quality assurance purposes, it is absolutely recommended to carry out pathological, radiological, and diagnostic work in specialized centers. PMID- 11482577 TI - Telomerase activity, Ki-67, cyclin D1 and A expression, and apoptosis in solitary fibrous tumors: additional features of a predictable course? AB - Solitary fibrous tumors (SFTs) are infrequent soft tissue neoplasms which are usually benign and surgically curable. However, their behavior is not always predictable, although several clinical and pathological criteria of malignancy have been established. In many cancers, including some soft tissue tumors, telomerase activity (TA) has been shown to be a new reliable pathological marker of malignancy. Overexpression of some cyclins is associated with higher degrees of malignancy and predictive of the clinical course. In this study, we evaluated TA, mitotic and apoptotic indices (MI, AI), and the expression of Ki-67, cyclins D1 and A in five typical and two clinicopathologically atypical SFTs, the latter two of which had also recurred. High TA was demonstrated in the two atypical cases, which also showed a higher labeling index to Ki-67, as well as higher cyclin D1 and A expression, and either none or very few apoptoses. We suggest that TA, Ki-67, cyclin expression, and AI be evaluated in SFTs as possible adjunctive pathological criteria of behavior. PMID- 11482578 TI - Matrix metalloproteinase-2 expression in laryngeal preneoplastic and neoplastic lesions. AB - Matrix metalloproteinase-2 (MMP-2), a member of gelatinases, is particularly important in the digestion of nonfibrillary and denaturated collagens; thus, it may play a role in tissue remodeling and in the invasion of malignant cells. The expression of MMP-2 has not yet been described in preneoplastic lesions of the larynx thus far. The purpose of this study was to evaluate whether the expression of MMP-2 plays a role in early laryngeal carcinogenesis. Laryngectomy specimens of 20 invasive carcinoma cases were studied. The slides with atypical hyperplasia, carcinoma in situ, and invasive carcinoma were selected from laryngectomy specimens. On these slides, 23 atypical hyperplasia and 17 carcinoma in situ areas were identified. MMP-2 expression was scored immunohistochemically on paraffin tissue sections using the avidin-biotin-peroxidase method. MMP-2 expression in all three groups was statistically different. A sequential increase in MMP-2 expression correlated significantly with the hypothesis of multistep carcinogenesis. In contrast, MMP-2 expression was not related to tumor stage, lymph node metastasis, or differentiation in squamous cell carcinomas. In conclusion, this sequential increase in MMP-2 expression points to an altered expression of MMP-2 in early neoplastic transformation in laryngeal mucosa, followed by an increasing expression during the progression of the disease. PMID- 11482579 TI - Bcl-2 correlates with tumor ploidy and nuclear morphology in early stage prostate carcinoma. A fine needle aspiration biopsy study. AB - We evaluated the nuclear morphology, ploidy, bcl-2 expression and in situ apoptosis in sections of fine-needle aspiration (FNA) biopsy specimens of thirty one randomly selected Stage B prostate carcinomas. Sections of paraffin-embedded pelleted cells obtained from FNA biopsy specimens were studied. Nuclear grade was determined according to the WHO system. Nuclear morphometry and DNA ploidy were carried out using an automated image analyzer. We used immunostaining and the TUNEL method to evaluate bcl-2 expression and in situ apoptosis. The median nuclear area increased with increasing nuclear grade. Ploidy analysis showed that 54.8% of tumors were diploid, 3.2% tetraploid and 41.9% aneuploid. Bcl-2 overexpression was found in 10 of 31 tumors. There was a significant positive correlation between bcl-2 expression and nuclear area (r(s): 0.45 p < 0.01). Nine of ten bcl-2-positive tumors had a nuclear area larger than the median of the series, and 70% of bcl-2-positive tumors were of the aneuploid type. The apoptotic index had a negative correlation with nuclear area, and the lowest indexes were found in aneuploid tumors. Bcl-2 expression showed a highly significant association with both parameters of high aggressiveness: nuclear size and aneuploidy. The combined evaluation of nuclear morphology, ploidy and cell survival parameters might better identify patients with poor prognosis among early stage prostate carcinomas diagnosed by FNA biopsies. PMID- 11482580 TI - Pituitary tumors in the elderly. AB - The present study was designed to investigate the incidence and immunohistochemical characteristics of pituitary tumors in the elderly. In our surgical collection of 1925 cases, we examined tumor tissue from 15 patients over 80 years of age. Pituitaries obtained at routine autopsies from 692 subjects over 80 years of age were also investigated. Of the 15 surgical cases studied, the majority of patients presented with chiasmatic syndromes, likely caused by macroadenomas. Gonadotroph adenomas were the most frequently diagnosed tumor type, followed by null-cell adenomas and oncocytomas. There is only one case with GH cell adenoma. Among 692 autopsy cases, 79 (11.4%) pituitaries were found to contain adenomas in the anterior lobe. In one pituitary, two separate adenomas were detected, hence the number of adenomas in our material was 80. All autopsy cases were microadenomas except one. The mean diameter of adenomas was 2.2 mm. ACTH cell adenomas were the most frequently diagnosed tumor type, followed by PRL cell adenomas and null cell adenomas. The occurrence of pituitary adenomas discovered after routine autopsy in the elderly was common, although these tumors were not found frequently in surgical cases over 80 years of age. Our immunohistochemical study revealed that many tumors contained one or more than one anterior pituitary hormone, although almost all pituitary adenomas were considered to be clinically inactive in surgical and autopsy cases. PMID- 11482581 TI - Tumoral calcium pyrophosphate dihydrate crystal deposition disease. A clinicopathologic analysis of five cases. AB - We describe five cases of tumoral calcium pyrophosphate dihydrate crystal deposition disease (CPPDCD) and discuss the clinical, radiological and pathological features. Patients included 4 males and 1 female, ranging in age from 49 to 70 years (median, 63 yrs). The wrist was involved in two patients. The thumb, palmar aspect of the proximal phalanx of the middle finger and dorsum of the carpal bone of the hand were involved in one patient each. In one patient, a preoperative diagnosis of chondrosarcoma had been made. Macroscopically, the lesion was a circumscribed whitish-gray mass with a more or less chalky appearance, measuring between 1.0 to 6.2 cm (median, 2.5 cm). Histologically, all five lesions contained areas of calcification with crystal deposits and chondroid metaplasia. The majority of crystals were rhomboid in shape, characteristic of CPPD, but some needle-shaped crystals were also identified, which resembled urate crystals. A review of the 54 reported cases of tumoral CPPDCD including our series indicated that they could be divided into two categories based on anatomic location: central (head and neck) type (n = 33) and distal (extremity) type (n = 21). Patients of these two groups were not different with respect to age and gender, but those with the central type often presented with a painful mass (15 patients, 46%), or neurological disturbances (11 patients, 33%). Patients with the distal type presented with a painless mass or swelling (12 patients, 57%), but none had neurological signs, although 8 (38.1%) presented with acute attack similar to tophaceous gout. Tumoral CP-PDCD should be differentiated from tophaceous gout, tumoral calcinosis, and malignant or benign tumors. PMID- 11482582 TI - Concurrent tubulovillous adenoma and transitional cell carcinoma associated with diffuse gastric and intestinal metaplasia of the defunctioned ureter. AB - Villous adenoma is a common lesion of the gastrointestinal tract, but it is rare in the ureter. Thus, as far as we know, only one case limited to this location has been described. Intestinal metaplasia of the urothelium is not rare. However, only one case of gastric metaplasia with pseudopyloric glands has been described in the literature. We here report in detail on a tubulovillous adenoma of the ureter associated with diffuse gastric and intestinal metaplasia and a concurrent primary, solid, high grade transitional cell carcinoma, with extensive clear cell change, in a 56-year-old male patient. He had undergone a left nephrectomy for renal tuberculosis twenty years earlier, and the lesions developed in the ureteric stump. To the best of our knowledge, such a combination of lesions has not been reported previously either in the ureter or in the rest of the urinary tract. The coexistence of diverse lesions in our case might represent the pluripotentiality of the urothelium in association with chronic inflammation and neoplastic induction. The present report also emphasizes the metaplastic and malignant potential of a defunctioned urothelial structure. This case is of particular interest, because these coexistent lesions arose simultaneously with an anatomically separate adenocarcinoma of the rectum (Dukes' B). The patient died 76 days after admission. The dismal prognosis of our case was determined by the advanced anatomical stage and the histological high grade of the transitional cell carcinoma of the ureter. PMID- 11482583 TI - Chordoid meningioma. A case report. AB - We report on a 62-year-old woman without Castleman's syndrome diagnosed with chordoid meningioma. A white, encapsulated brain tumor was located in the parietal lobe of the left hemisphere of the cerebrum, adhered to the dura, and was separated from the cerebrum. The tumor revealed a multilobular arrangement of two types of neoplastic cells, and the surrounding myxoid stroma was separated by incomplete fibrous septa. Neoplastic cells consisted of myxomatous and meningothelial cells. The former made up about four-fifths of the tumor, had a vacuolar cytoplasm, and were arranged in a chordoma-like cord pattern. They were floating in myxoid stroma. The latter had an eosinophilic spindle or epithelioid cytoplasm and were disposed in lobules. Coarse eosinophilic materials positive for periodic acid-Schiff stain were deposited among them. Transitional cells between two types of cells were also observed. Both neoplastic cells were positive for vimentin and Leu-7 (CD57) in their cytoplasm, and were consistently negative for epithelial membrane antigen, S-100 protein, and cytokeratin. PMID- 11482584 TI - Creating more effective health plan quality reports for consumers: lessons from a synthesis of qualitative testing. AB - OBJECTIVE: Social marketing techniques such as consumer testing have only recently been applied to develop effective consumer health insurance information. This article discusses lessons learned from consumer testing to create consumer plan choice materials. DATA SOURCES/STUDY SETTING: Data were collected from 268 publicly and privately insured consumers in three studies between 1994 and 1999. STUDY DESIGN: Iterative testing and revisions were conducted to design seven booklets to help Medicaid, Medicare, and employed consumers choose a health plan. DATA COLLECTION METHODS: Standardized protocols were used in 11 focus groups and 182 interviews to examine the content, comprehension, navigation, and utility of the booklets. PRINCIPAL FINDINGS: A method is suggested to help consumers narrow their plan choices by breaking down the process into smaller decisions using a set of guided worksheets. CONCLUSION: Implementing these lessons is challenging and not often done well. This article gives examples of evidence-based approaches to address cognitive barriers that designers of consumer health insurance information can adapt to their needs. PMID- 11482585 TI - Reporting of CAHPS quality information to medicare beneficiaries. AB - OBJECTIVES: To assess which Consumer Assessment of Health Plans (CAHPS) survey measures Medicare beneficiaries find the most meaningful, how beneficiaries and information intermediaries interpret different formats for presenting CAHPS information, and how beneficiaries have reacted to the CAHPS information included in the annual mailing to beneficiaries called Medicare & You 2000. DATA SOURCES: Fourteen focus groups of beneficiaries and State Health Insurance Assistance Program counselors, more than 200 cognitive interviews, and 122 mall-intercept interviews with beneficiaries were conducted from spring 1998 through winter 2000. STUDY DESIGN: In 1998 focus groups and cognitive interviews were conducted with Medicare beneficiaries and State Health Insurance Assistance Program counselors to determine which CAHPS measures to report to Medicare beneficiaries and how to report this information. In 1999 additional focus groups and mall intercept interviews were conducted to determine which measures to include in Medicare & You 2000. To obtain feedback on the CAHPS information in Medicare & You 2000 additional focus groups were conducted in winter 2000. PRINCIPAL FINDINGS: Focus group participants indicated that getting the care they need quickly, having access to specialists, and communicating well with doctors were more important to them than nonmedical characteristics of plans. Most beneficiaries had problems interpreting quality information. Many misinterpreted star charts, and while bar charts appear easier to read, many beneficiaries still had trouble interpreting the information on these charts. Most beneficiaries did not consider quality information important to them and most were unaware of the availability of CAHPS information. CONCLUSIONS: Many challenges lie ahead in making quality information meaningful to Medicare beneficiaries. These challenges include increasing awareness of the existence of this information, educating beneficiaries about how this information can help in choosing a health plan, continuing to simplify reporting formats, assuring beneficiaries that this information comes from a credible source, and providing guidance to beneficiaries about how quality information can help with health care decisions. PMID- 11482586 TI - Differences in the structure of CAHPS measures among the medicare fee-for service, medicare managed care, and privately insured populations. AB - OBJECTIVE: To confirm in a new population, the Medicare fee-for-service population, the factor structure previously found in two Consumer Assessment of Health Plans Study (CAHPS) field-test surveys with Medicare HMO and adult privately insured populations. DATA SOURCES: Primary data were collected in the fall of 1998. Survey responses from the Medicare Fee-for-Service CAHPS survey field test were compared to results from the Medicare HMO and adult privately insured field-test studies conducted in the fall of 1996. STUDY DESIGN: Respondents for the field-test survey were a random sample of Medicare beneficiaries in five states who had opted for the original Medicare plan (fee for-service). DATA COLLECTION: Data were collected by a mailed survey with a telephone follow-up survey to those who did not return the mailed survey. PRINCIPAL FINDINGS: A confirmatory factor analysis in two different samples of Medicare fee-for-service beneficiaries provided basic support for a previously reported three-factor structure underlying the CAHPS reports and rating items: (1) quality of provider or staff communications; (2) timely access to quality health care; and (3) quality of plan administration. An exploratory factor analysis revealed a variant three-factor structure. CONCLUSION: Because of differences in the factor structures among the different populations discussed, caution needs to be exercised in any composite development, based on factor analysis or any other basis, by which cross-population comparisons will be made. Comparisons should only be made on composites representing stable structure across all populations concerned. PMID- 11482587 TI - Do employees use report cards to assess health care provider systems? AB - OBJECTIVE: To investigate consumers' use of report cards that provide information on service quality and satisfaction at the provider group level. DATA SOURCES: In 1998 we conducted a telephone survey of randomly selected employees in firms aligned with the Buyers Health Care Action Group (BHCAG) in the Minneapolis-St. Paul market. STUDY DESIGN: Univariate probit models were used to determine report card utilization, perceived helpfulness of the report card, and ease of selecting a provider group. The characteristics used in the models included health status, age, gender, education, residency, job tenure, marital status, presence of dependent children, household income, and whether consumers changed provider groups. DATA COLLECTION: Our sample consists of survey responses from 996 single individuals (a response rate of 91 percent) and 913 families (a response rate of 96 percent). The survey was supplemented with data obtained directly from employers aligned with BHCAG. PRINCIPLE FINDINGS: Consumers who changed to a new provider group are more likely to use report card information and find it helpful, consumers employed in large firms are less likely to use the report card, and families who use information from their own health care experiences are less likely to find the report card helpful. In addition, individuals who changed to a new provider group are more likely to find the selection decision difficult. CONCLUSION: The findings show that health care consumers are using satisfaction and service-quality information provided by their employers. PMID- 11482588 TI - Health plan decision making with new medicare information materials. AB - OBJECTIVE: To examine the effect of providing new Medicare information materials on consumers' attitudes and behavior about health plan choice. DATA SOURCE: New and experienced Medicare beneficiaries who resided in the Kansas City metropolitan statistical area during winter 1998-99 were surveyed. More than 2,000 computer-assisted telephone interviews were completed across the two beneficiary populations with a mean response rate of 60 percent. STUDY DESIGN: Medicare beneficiaries were randomly assigned to a control group or one of three treatment groups that received varying amounts and types of new Medicare information materials. One treatment group received the Health Care Financing Administrations's pilot Medicare & You 1999 handbook, a second group received the same version of the handbook and a Medicare version of the Consumer Assessment of Health Plans (CAHPS) report, and a third treatment group received the Medicare & You bulletin, an abbreviated version of the handbook. PRINCIPAL FINDINGS: Results of the study suggest that the federal government's new consumer information materials are having some influence on Medicare beneficiaries' attitudes and behaviors about health plan decision making. Experienced beneficiary treatment group members were significantly more confident with their current health plan choice than control group members, but new beneficiaries were significantly less likely to use the new materials to choose or change health plans than control group members. In general the effects on confidence and health plan switching did not vary across the different treatment materials. CONCLUSIONS: The 1999 version of the Medicare & You materials contained a message that it is not necessary to change health plans. This message appears to have decreased the likelihood of using the new materials to choose or change plans, whereas other materials to which beneficiaries are exposed may encourage plan switching. Because providing more information to beneficiaries did not result in commensurate increases in confidence levels or rate of health plan switching, factors other than the amount of information, such as how the information is presented, may be more critical than volume. PMID- 11482589 TI - Case-mix adjustment of the National CAHPS benchmarking data 1.0: a violation of model assumptions? AB - OBJECTIVE: To compare models for the case-mix adjustment of consumer reports and ratings of health care. DATA SOURCES: The study used the Consumer Assessment of Health Plans (CAHPS) survey 1.0 National CAHPS Benchmarking Database data from 54 commercial and 31 Medicaid health plans from across the United States: 19,541 adults (age > or = 18 years) in commercial plans and 8,813 adults in Medicaid plans responded regarding their own health care, and 9,871 Medicaid adults responded regarding the health care of their minor children. STUDY DESIGN: Four case-mix models (no adjustment; self-rated health and age; health, age, and education; and health, age, education, and plan interactions) were compared on 21 ratings and reports regarding health care for three populations (adults in commercial plans, adults in Medicaid plans, and children in Medicaid plans). The magnitude of case-mix adjustments, the effects of adjustments on plan rankings, and the homogeneity of these effects across plans were examined. DATA EXTRACTION: All ratings and reports were linearly transformed to a possible range of 0 to 100 for comparability. PRINCIPAL FINDINGS: Case-mix adjusters, especially self-rated health, have substantial effects, but these effects vary substantially from plan to plan, a violation of standard case-mix assumptions. CONCLUSION: Case-mix adjustment of CAHPS data needs to be re-examined, perhaps by using demographically stratified reporting or by developing better measures of response bias. PMID- 11482590 TI - Racial and ethnic differences in parents' assessments of pediatric care in Medicaid managed care. AB - OBJECTIVE: This study examines whether parents' reports and ratings of pediatric health care vary by race/ethnicity and language in Medicaid managed care. DATA SOURCES: The data analyzed are from the National Consumer Assessment of Health Plans (CAHPS) Benchmarking Database 1.0 and consist of 9,540 children enrolled in Medicaid managed care plans in Arkansas, Kansas, Minnesota, Oklahoma, Vermont, and Washington state from 1997 to 1998. DATA COLLECTION: The data were collected by telephone and mail, and surveys were administered in Spanish and English. The mean response rate for all plans was 42.1 percent. STUDY DESIGN: Data were analyzed using multiple regression models. The dependent variables are CAHPS 1.0 ratings (personal doctor, specialist, health care, health plan) and reports of care (getting needed care, timeliness of care, provider communication, staff helpfulness, plan service). The independent variables are race/ethnicity (white, African American, American Indian, Asian, and Hispanic), Hispanic language (English or Spanish), and Asian language (English or other), controlling for gender, age, education, and health status. PRINCIPAL FINDINGS: Racial/ethnic minorities had worse reports of care than whites. Among Hispanics and Asians language barriers had a larger negative effect on reports of care than race/ethnicity. For example, while Asian non-English-speakers had lower scores than whites for staff helpfulness (beta = -20.10), timeliness of care (beta = 18.65), provider communication (beta = -17.19), plan service (beta = -10.95), and getting needed care (beta = -8.11), Asian English speakers did not differ significantly from whites on any of the reports of care. However, lower reports of care for racial/ethnic groups did not translate necessarily into lower ratings of care. CONCLUSIONS: Health plans need to pay increased attention to racial/ethnic differences in assessments of care. This study's finding that language barriers are largely responsible for racial/ethnic disparities in care suggests that linguistically appropriate health care services are needed to address these gaps. PMID- 11482591 TI - Differences in CAHPS adult survey reports and ratings by race and ethnicity: an analysis of the National CAHPS benchmarking data 1.0. AB - OBJECTIVE: To examine racial/ethnic group differences in adults' reports and ratings of care using data from the National Consumer Assessment of Health Plans (CAHPS) survey Benchmarking Database (NCBD) 1.0. DATA SOURCE: Adult data from the NCBD 1.0 is comprised of CAHPS 1.0 survey data from 54 commercial and 31 Medicaid health plans from across the United States. A total of 28,354 adult respondents (age > or = 18 years) were included in this study. Respondents were categorized as belonging to one of the following racial/ethnic groups: Hispanic (n = 1,657), white (n = 20,414), black or African American (n = 2,942), Asian and Pacific Islander (n = 976), and American Indian or Alaskan native (n = 588). STUDY DESIGN: Four single-item global ratings (personal doctor, specialty care, overall rating of health plan, and overall rating of health care) and five multiple-item report composites (access to needed care, provider communication, office staff helpfulness, promptness of care, and health plan customer service) from CAHPS 1.0 were examined. Statistical Analyses. Multiple regression models were estimated to assess differences in global ratings and report composites between whites and members of other racial/ethnic groups, controlling for age, gender, perceived health status, educational attainment, and insurance type. PRINCIPAL FINDINGS: Members of racial/ethnic minority groups, with the exception of Asians/Pacific Islanders, reported experiences with health care similar to those of whites. However, global ratings of care by Asians/Pacific Islanders are similar to those of whites. CONCLUSIONS: Improvements in quality of care for Asians/Pacific Islanders are needed. Comparisons of care in racially and ethnically diverse populations based on global ratings of care should be interpreted cautiously. PMID- 11482592 TI - The role of performance measures for improving quality in managed care organizations. AB - OBJECTIVES: To understand how managed care plans use performance measures for quality improvement and to identify the strengths and weaknesses of currently used standardized performance measures such as the Health Plan Employer Data and Information Set (HEDIS) and the Consumer Assessment of Health Plans (CAHPS) survey. DATA SOURCES/STUDY SETTING: Representatives (chief executive officers, medical directors, and quality-improvement directors) from 24 health plans in four states were surveyed. The overall response rate was 58.3 percent, with a mean of 1.8 respondents per plan. STUDY DESIGN: This exploratory qualitative research used a purposive sample of respondents. Two study authors conducted separate one-hour tape-recorded telephone interviews with multiple respondents from each health plan. PRINCIPAL FINDINGS: All managed care organizations interviewed use performance measures for quality improvement but the degree and sophistication of use varies. Many of our respondent plans use performance measures to target quality-improvement initiatives, evaluate current performance, establish goals for quality improvement, identify the root cause of problems, and monitor performance. CONCLUSION: Performance measures are used for quality improvement in addition to informing external constituents, but additional research is needed to understand how the benefits of measurement can be maximized. PMID- 11482593 TI - The effects of disseminating performance data to health plans: results of qualitative research with the Medicare Managed Care plans. AB - OBJECTIVE: To assess the information needs and responses of managed care plans to the Medicare Managed Care Consumer Assessment of Health Plans Study (MMC-CAHPS). DATA SOURCES/STUDY SETTING: One hundred sixty-five representatives of Medicare managed care plans participated in focus groups or interviews in the spring of 1998, 1999, and 2000. STUDY DESIGN: In 1998 focus groups were conducted with representatives of managed care plans to develop and test a print report of MMC CAHPS results. After the reports were disseminated focus groups and interviews were conducted in 1999 and 2000 to identify perceptions, uses, and potential enhancements of the report. DATA COLLECTION/EXTRACTION METHODS: The study team conducted a total of 23 focus groups and 12 telephone interviews and analyzed the transcripts to identify major themes. PRINCIPAL FINDINGS: In 1998 participants identified the report content and format that best enabled them to assess their performance relative to other Medicare managed care plans. In 1999 and 2000 participants described their responses to and uses of the report. They reported comparing the MMC-CAHPS results to internal surveys and presenting the results to senior managers, market analysts, and quality-improvement teams. They also indicated that the report's usefulness would be enhanced if it were received within six months of survey completion and if additional data analysis was presented. CONCLUSIONS: Focus group results suggest that the MMC-CAHPS report enhances awareness and knowledge of the comparative performance of Medicare managed care plans. However, participants reported needing additional analysis of survey results to target quality-improvement activities on the populations with the most reported problems. PMID- 11482594 TI - Postweaning multisystemic wasting syndrome induced after experimental inoculation of cesarean-derived, colostrum-deprived piglets with type 2 porcine circovirus. AB - Cesarean-derived, colostrum-deprived pigs (n = 23) were inoculated intranasally and subcutaneously with a low cell culture passage of type 2 porcine circovirus. In 11 pigs, a persistent fever that lasted 7-17 days began 12-15 days after inoculation with virus. Additional signs of disease in those 11 pigs included depression (11 of 11 pigs), palpable enlargement of inguinal, prefemoral, and popliteal lymph nodes (11 of 11), icterus (6 of 11), and hyperpnea (2 of 11). The remaining 12 pigs had fever that occurred intermittently for 2-4 days between days 12 and 20 postinoculation. Overt signs of disease in those pigs were limited to palpable enlargement of inguinal and popliteal lymph nodes (9 of 12 pigs). When compared with control pigs of similar age, the average daily rate of weight gain for all pigs inoculated with virus was less over a 2-week period that began 2 weeks post inoculation. At postmortem examination, lymph node enlargement was seen in 14 of 14 pigs euthanized between days 20 and 28 postinoculation. Lymph node enlargement was especially prominent in pigs that developed a persistent fever. Microscopic lesions noted in pigs that developed a persistent fever included cellular depletion in lymphoid tissues; hepatic cell necrosis; and lymphogranulomatous inflammation of lymph nodes, Peyer's patches of the intestine, liver, kidney, and heart. Virus was isolated with varying frequency from nasal, rectal, or tonsil swab specimens, buffy coat, serum, urine, and lung lavage fluid obtained antemortem or postmortem. Virus was isolated from or viral DNA was detected in a variety of tissues obtained postmortem up to 125 days postinoculation. Antibody against type 2 porcine circovirus usually was detected in serum between 15 and 20 days postinoculation; however, antibody against virus was not detected in serum from 4 pigs euthanized 20-24 days postinoculation. Direct contact with pigs inoculated with virus 42 days previously resulted in transmission of virus to 3 of 3 control pigs. PMID- 11482595 TI - A new serotype adenovirus isolated from a goat in the United States. AB - A virus (T94-0353) isolated from the small intestine of a 3-week-old kid with diarrhea and serous ocular and nasal discharge was identified as an adenovirus based on morphologic and physicochemical characteristics. Neutralization tests and restriction endonuclease analysis comparing the caprine adenovirus with the prototype bovine and ovine adenovirus serotypes and a previously isolated caprine adenovirus showed that the caprine isolate was antigenically distinct, produced a unique restriction pattern compared with currently recognized bovine, caprine, and ovine adenoviruses, and represents a new adenovirus type. The role and significance of naturally acquired adenovirus infection in respiratory and enteric disease in goats has not been established. Isolation of adenovirus from goats with disease coupled with seroepidemiologic and pathogenicity studies will help define the role of the adenoviruses in disease production. PMID- 11482596 TI - PCR-based detection of an emerging avian pneumovirus in US turkey flocks. AB - Avian pneumovirus (APV) or turkey rhinotracheitis virus (TRTV) is an important respiratory pathogen of domesticated poultry in many countries in Europe, Africa, and Asia. Until recently, the United States was considered free of APV. In late 1996, an atypical upper respiratory tract infection appeared in turkey flocks in Colorado and shortly thereafter in turkey flocks in Minnesota. An avian pneumovirus (APV-US) that was serologically distinct from the previously described TRTV was isolated as the primary cause of the new syndrome. The nucleotide sequence of a fragment of the APV-US fusion gene was determined and used to develop a polymerase chain reaction-based assay that specifically detects APV-US viral nucleic acid sequences in RNA extracts of tracheal swabs and turbinate homogenates. The assay is highly sensitive in that it can detect <0.01 TCID50 of APV. The availability of this assay enables the rapid and accurate determination of APV-US in infected poultry flocks. PMID- 11482597 TI - Spatial and temporal clustering of Salmonella serotypes isolated from adult diarrheic dairy cattle in California. AB - Electronic records from the California Animal Health and Food Safety Laboratory System between January 1, 1991, and June 30, 1998, were reviewed, and data on 593 Salmonella serotypes isolated from feces or intestinal contents of adult dairy cattle with diarrhea were analyzed to determine spatial and temporal clustering. A statistically significant clustering in space was detected with 10 (Typhimurium, Montevideo, Muenster, Anatum, Give, Menhaden, Kentucky, Agona, Derby, and Newport) of the 13 serotypes examined. Significant temporal clustering was also detected with 10 serotypes (Typhimurium, Montevideo, Muenster, Anatum, Give, Newbrunswick, Menhaden, Kentucky, Derby, and Newport). Six serotypes (Anatum, Menhaden, Montevideo, Muenster, Newbrunswick, and Newport) were significantly spatially and temporally clustered. A difference in temporal and spatial distribution patterns of some serotypes associated with diarrhea in adult dairy in California was found. Knowledge of the specific type of clustering, if present, should improve our understanding of the transmission and control of salmonellosis in the field. PMID- 11482598 TI - Isotype-specific antibody responses of cattle to Salmonella Dublin lipopolysaccharide and porin following Salmonella Dublin vaccination and acute and chronic infection. AB - Stimulation of different T-cell subsets during antigen presentation influences the antibody isotype response to an antigen. Salmonella infection and Salmonella bacterin vaccination are likely to stimulate different T-cell subtypes. The objective of this study was to determine whether there are differences in the isotype response of cattle to Salmonella antigens following Salmonella infection and Salmonella bacterin vaccination. Sera from Salmonella bacterin-vaccinated, experimentally infected, and chronically infected (carrier) adult cattle collected during previous studies was used to evaluate the IgG1, IgG2, and IgM isotype responses of cows to Salmonella serotype Dublin lipopolysaccharide (LPS) and porin. Following vaccination and experimental oral infection, IgG1 titers to LPS and porin rose more quickly and persisted longer than did IgG2 titers. In contrast to Salmonella infection, bacterin vaccination stimulated a weak response to Salmonella porin. Salmonella infection also induced a higher IgG2:IgG1 titer ratio to LPS than did bacterin vaccination. Chronic Salmonella infection induced the highest LPS and porin IgG2:IgG1 titer ratios and the highest correlation between LPS and porin titers. Response operating characteristic curves for each isotype-specific enzyme-linked immunosorbent assay (ELISA) were determined to evaluate the effect of isotype on the sensitivity and specificity of Salmonella ELISA serology for distinguishing sera of Salmonella carriers from those of vaccinated and acutely infected cows. IgG2 titers to LPS and porin provide a more specific indicator of chronic Salmonella infection status than do IgG1 titers to the same antigens with little to no loss in sensitivity. PMID- 11482599 TI - Differential detection of Bartonella species and strains in cat scratch disease diagnostics by polymerase chain reaction amplification of 16S ribosomal RNA gene. AB - Cat scratch disease (CSD) has been difficult to diagnose in animals because of the protracted clinical course of infection and the quiescent phases when the microbial culprit lies dormant. The causative agent in CSD appears to be multiple species and strains of Bartonella. Using polymerase chain reaction (PCR) techniques for amplification of highly variable regions of the 16S ribosomal RNA (rRNA) gene sequence, a very sensitive species- and strain-specific assay for CSD causing Bartonella species was developed. PCR primers were designed to specifically amplify the 16S rRNA gene of Bartonella species but not of other microbial pathogens. This initial PCR was multiplexed with a universal primer set, based on conserved sequence regions in the 16S rRNA gene, that provides a 162-bp fragment in all species tested. Subsequently, 3 distinct nested PCR primer sets enabled the individual amplification and specific detection of Bartonella henselae type 1, B. henselae type II, and B. clarridgeae. Thus, this 2-step PCR procedure enabled the sensitive detection and identification of these species and the B. henselae genotype by exploiting minor sequences differences. Verification of these results were demonstrated with both sequencing and ligase chain reaction techniques. The diagnostic usefulness of this CSD test has been demonstrated by the analysis of specimens from control and infected cats. The diagnosis was confirmed and the specific B. henselae strain was correctly identified in peripheral blood specimens obtained from control and strain-specific CSD-infected cats. Such an accurate and sensitive diagnostic tool for the detection and identification of CSD causative agents should be a useful for the medical, veterinary, and scientific communities. PMID- 11482600 TI - Semiquantitative determination of ergot alkaloids in seed, straw, and digesta samples using a competitive enzyme-linked immunosorbent assay. AB - Ergot alkaloids present in endophyte-infected (E+) tall fescue cause fescue toxicosis and other toxic effects in livestock that consume infected plant tissue, leading to significant financial losses in livestock production each year. The predominant method currently in use for quantifying ergot alkaloid content in plant tissue is through high-performance liquid chromatography (HPLC), which quantifies the amount of ergovaline, one of many ergot alkaloids in E+ plant tissue. The enzyme-linked immunosorbent assay (ELISA) method used in this study detects quantities of nonspecific ergot alkaloids and therefore accounts for greater amounts of the total ergot alkaloid content in E+ tissue than does HPLC. The ELISA can also be used to more expediently analyze a larger number of forage samples without sophisticated and costly analytical equipment and therefore could be more desirable in a diagnostic setting. The purpose of this study was to evaluate the between-day and within-run variability of the ELISA and to determine the binding efficiency of 6 ergot alkaloids to the 15F3.E5 antibody used in the competitive ELISA to ascertain its feasibility as a quick analysis tool for ergot alkaloids. Straw samples had an average coefficient of variation (CV) for concentration of 10.2% within runs and 18.4% between runs, and the seed samples had an average CV for concentration of 13.3% within runs and 24.5% between runs. The grass tissue-based lysergic acid standard curve calculated from the ELISA had an average r2 of 0.99, with a CV of 2.1%. Ergocryptine, ergocristine, ergocornine, and ergotamine tartrate did not bind strongly to the 15F3.E5 antibody because of the presence of large side groups on these molecules, which block their binding to the antibody, whereas ergonovine and ergonovine maleate were bound much more efficiently because of their structural similarity to lysergic acid. Clarified rumen fluid was tested as an additional matrix for use in the ergot alkaloid competitive ELISA to determine whether future livestock metabolism experiments on the postingestion fate of ergot alkaloids in ruminants could utilize this assay as a quick screening tool for the presence of nonspecific ergot alkaloids in rumen fluid. HPLC and ELISA procedures were compared for their ability in determining ergot alkaloid toxicity based on the repeatability of the procedures and on the specific compounds they measure. The ratio of ELISA concentration to HPLC concentration (ergovaline) varied from 2.00 to 2.81 in seed samples and from 0.62 to 8.66 in straw samples, showing no consistent pattern between the 2 methods. Based on the lack of data at present for the identity of the toxin causing endophyte toxicosis and the lack of agreement between the ergovaline HPLC and ELISA analyses for ergot alkaloids, each method is equally valid as an indicator of toxicityand is the best means for determining the quantity of the specific toxin(s) they measure. PMID- 11482601 TI - Foxtail-induced ulcerative stomatitis outbreak in a Missouri stable. AB - Twenty of 25 horses in a well-managed Missouri boarding stable were diagnosed with gingivitis/stomatitis. Gross examination of the affected horses revealed varying degrees of gingivitis ranging from mild periodontal swelling to marked swelling and erythema with ulceration and hemorrhage. Fine hair-like material was embedded within the intensely affected areas. Gingival biopsies from 4 affected horses contained pyogranulomatous inflammation with, in some cases, numerous eosinophils and several grass awns in cross and longitudinal section. Numerous foxtail seed heads were identified in hay samples. Examination of the records revealed that all of the affected horses had been fed the suspect hay, with the exception of 1 horse. Although not deliberately fed the suspect hay, this horse did have access to the hay when turned out into the exercise paddock. The lesions resolved following a change in hay source. PMID- 11482602 TI - Salpingitis in Pekin ducks associated with concurrent infection with Tetratrichomonas sp. and Escherichia coli. AB - Increased mortality (1.5% per week) and low egg production (5-10% lower than normal) were observed in a flock of domestic breeding Pekin ducks (Anas platyrhynchos). At necropsy, salpingitis and peritonitis were the most significant findings. Histologically, there was accumulation of necrotic debris in the lumen of the oviduct. Numerous bacteria and trichomonads were observed histologically in the lumen of the vagina and occasionally in the shell gland. Escherichia coli and a trichomonad were isolated from the oviduct. The trichomonads were oval (6-8 microm long, 4.5-6 microm wide) and had 4 anterior flagella and an undulating membrane extending over the entire length of the body, finishing in a long posterior flagellum. Morphology was consistent with trichomonads of the genus Tetratrichomonas. Comparative sequence analysis of the 5.8S ribosomal RNA gene and the flanking internal transcribed space regions of the trichomonad isolate did not closely match with available sequences of the same region of other trichomonadid protozoa. PMID- 11482603 TI - An outbreak of toxoplasmosis in farmed mink (Mustela vison S.). AB - A large Wisconsin mink (Mustela vison) farm experienced an outbreak of toxoplasmosis in the spring of 1999 following an outbreak of canine distemper during the previous fall. Major clinical signs for pregnant females included reduced feed consumption, abortions, and stillborn kits; kits < or =3 weeks old experienced ataxia and mortality. Of 7,800 females, 1,976 (26%) lost their entire litter either from abortion or neonatal mortality. Kit mortality from 7 days to 3 weeks of age was 3,300, and overall kit mortality attributed to the toxoplasmosis outbreak was 10,408. Six neonatal mink kits, 12 3-week-old kits, and 2 adult female mink were submitted to the University of Minnesota Veterinary Diagnostic Laboratory for diagnostic workup. Gross postmortem lesions were limited to empty stomachs (12 of 12 kits) and pale livers (4 of 12 kits) for the 3-week-old kits. Major microscopic lesions included interstitial pneumonia, encephalitis, encephalomalacia, and myocarditis. Toxoplasmosis was diagnosed by microscopic lesions, microscopic lesion distribution, and the detection of Toxoplasma gondii tachyzoites by immunohistochemistry. This is the first detailed report of an outbreak of toxoplasmosis in mink in the United States. PMID- 11482604 TI - Validation of a competitive enzyme-linked immunosorbent assay for diagnosing Babesia equi infections of Moroccan origin and its use in determining the seroprevalence of B. equi in Morocco. AB - A highly specific and sensitive competitive enzyme-linked immunosorbent assay for detection of specific antibody to Babesia equi in serum from equids was validated for use in Morocco. The assay is based on the specific inhibition of binding of a monoclonal antibody to a conserved epitope within a recombinant parasite peptide by serum from infected animals. The assay was compared to an established indirect immunofluorescence assay, with a concordance of 91%. The assay was used to determine seroprevalence for B. equi infections in donkeys and horses throughout Morocco. A total of 578 sera (163 horses and 415 donkeys) from 6 locations representing different bioclimatic regions were assayed. An analysis of variance, indicated no significant effect of location; however, donkeys were significantly more likely than horses to be seropositive. Management conditions contribute to greater tick infestations and thus Babesia exposure in donkeys than in horses. PMID- 11482605 TI - Cutaneous neosporosis in two adult dogs on chronic immunosuppressive therapy. AB - Antemortem diagnosis of generalized ulcerative and pyogranulomatous dermatitis with numerous intralesional tachyzoites was made from skin biopsy specimens from 2 adult dogs on chronic immunosuppressive therapy. A 9-year-old Italian Greyhound was on long-term corticosteroid therapy for the treatment of a lupus-like systemic autoimmune disorder, and a 7-year-old Labrador Retriever had received several months of chemotherapy for lymphosarcoma. The tachyzoites were identified as Neospora caninum by immunoperoxidase immunohistochemistry. Both dogs were treated with clindamycin. Lesions in the Greyhound resolved; however, the Labrador Retriever was euthanized because of evidence of neuromuscular disease, despite improvement of the skin lesions. These 2 cases indicate that cutaneous neosporosis can occur in adult dogs on chronic immunosuppressive therapy. The disease may result from reactivation of a congenital infection and/or a recently acquired primary infection. PMID- 11482606 TI - Septicemia associated with Stenotrophomonas maltophilia in a West African dwarf crocodile (Osteolaemus tetraspis subsp. tetraspis). AB - A 17-year-old male captive West African dwarf crocodile (Osteolaemus tetraspis subsp. tetraspis) died 1 month after fighting with a penmate. Abrasions were present on the head and mandible. Necropsy revealed a vegetative valvular lesion of the left atrioventricular valve, miliary foci of necrosis in the endocardium and myocardium, multiple duodenal and rectal ulcers, and serous atrophy of body fat. Stenotrophomonas maltophilia was isolated in pure culture from lung, liver, and kidney. Gram-negative bacilli were seen histologically in the valvular lesion and in foci of necrosis in the myocardium, liver, spleen, pancreas, kidney, and intestine. Septic thrombi in multiple tissues, arteritis, and pneumonia were additional histologic lesions. Findings indicated that the crocodile died from acute S. maltophilia septicemia, although the primary site of infection was not determined. Stenotrophomonas maltophilia is ubiquitous in the environment and is recognized as an important nosocomial pathogen in humans. PMID- 11482607 TI - Comparison of GN Hajna and tetrathionate as initial enrichment for salmonellae recovery from swine lymph nodes and cecal contents collected at slaughter. AB - An epidemiologic survey was conducted to determine the prevalence of salmonellae in swine from 5 farms of an integrated swine operation. The purpose of this study was to evaluate the recovery efficiencies for salmonellae from swine lymph nodes and cecal contents when GN Hajna and tetrathionate were compared as initial enrichments. Salmonellae were isolated from 61% of 645 pigs at slaughter; 324 positive cultures were from lymph nodes, and 224 were from cecal contents. Frequently, pigs had salmonellae isolated from both the lymph nodes and cecal contents. Total isolations, regardless of source, were similar for GN Hajna (247) and tetrathionate (301). There was no difference (P > 0.05) in the number of isolations from lymph nodes when GN Hajna enrichment was compared with tetrathionate enrichment (174 vs. 150). However, there was a significant (P < 0.05) advantage of utilizing tetrathionate when compared with GN Hajna for isolations from cecal contents (151 vs. 73). PMID- 11482608 TI - Evaluation of a new sandwich enzyme-linked immunosorbent assay for detection of bovine viral diarrhea virus in unprocessed fetal bovine serum. AB - A new sandwich enzyme-linked immunosorbent assay (S-ELISA) kit that uses raw (unprocessed) fetal bovine serum (FBS) as the testing sample was evaluated for upstream bovine viral diarrhea virus (BVDV) testing. Pooled FBS samples (n = 84) were tested using the S-ELISA. Thirty serum samples originating from persistently infected (PI) calves that had been confirmed by virus isolation (VI) as BVDV positive and another 30 samples previously confirmed by VI as BVDV negative were also evaluated. Of the 84 field samples, the S-ELISA detected 13 (15.5%) BVDV positive specimens. When these 13 positive samples were tested by VI and immunofluorescent assay, 11 (84.6%) were positive and 2 (15.4%) were negative. The S-ELISA was positive for all 30 PI samples (100%) and negative for all 30 negative samples (100%). These data indicate that the new kit is a relatively reliable diagnostic tool and can be considered for upstream detection of BVDV contaminated raw FBS pools. PMID- 11482609 TI - Multiplex polymerase chain reaction for distinguishing Taylorella equigenitalis from Taylorella equigenitalis-like organisms. AB - It is difficult to distinguish isolates of Taylorella equigenitalis, the cause of contagious equine metritis, from a T. equigenitalis-like organism isolated from asymptomatic donkeys and horses. Although T. equigenitalis is responsible for a severe, contagious disease of the reproductive tract of equids, the T. equigenitalis-like organism, although contagious, does not appear to produce disease. Because of the economic consequences of correctly distinguishing isolates of these 2 microorganisms, a polymerase chain reaction (PCR)-based assay was developed that will distinguish isolates of T. equigenitalis from the T. equigenitalis-like microorganism. The primers used in the PCR assay were designed to amplify unique regions of the gene encoding the 16S ribosomal RNA. PMID- 11482610 TI - Causes of caprine abortion: diagnostic assessment of 211 cases (1991-1998). AB - In an 8-year period, 1991-1998, 217 accessions of caprine abortions were submitted to the California Veterinary Diagnostic Laboratory System. Of these 217 submissions, 211 were suitable for examination in this study (6 had insufficient data). Infectious agents as the cause of abortions were found in 37% of the cases: bacterial agents were identified in 30.5%, viral agents in 2%, fungal agents in 0.5%, and protozoal agents in 4% of the cases submitted. The most common causes of abortions were Chlamydia psittaci and Coxiella burnetii infection, which accounted for 23% of all goat abortions. Mineral deficiencies were observed in 4%, fetal anomalies accounted for 3%, and leukoencephalomalacia of the brain (probable oxygen deprivation) accounted for 3% of the submissions. No diagnosis was made in 112 of the 211 submissions (53%). No lesions were noted in 104 of the submissions (49%). The other 8 submissions (4%) had histologic lesions suggestive of a bacterial agent; however, no infectious agents were identified in these cases. PMID- 11482611 TI - Hepatic hyaline globules in an Eclectus parrot (Eclectus roratus). AB - Hepatic hyaline globules, similar to those reported in some human livers, were observed in liver tissue from an Eclectus parrot (Eclectus roratus). The cytoplasmic inclusions were periodic acid-Schiff positive and diastase resistant and failed to stain by acid-fast or Congo red techniques. Ultrastructurally, the hepatic globules were composed of granular amorphous material with small peripheral striations that extended into the cytoplasm. PMID- 11482612 TI - Use of eosin methylene blue agar to differentiate Escherichia coli from other gram-negative mastitis pathogens. PMID- 11482614 TI - More on errors in the surgical practice, and what can be done about it. PMID- 11482613 TI - Comparison of porcine reproductive and respiratory syndrome virus growth in media supplemented with fetal bovine serum or a serum replacement. AB - Commercially available serum replacements are often used in cell culture as a cheaper and less variable substitute for fetal bovine serum (FBS). The growth of porcine reproductive and respiratory syndrome virus (PRRSV) isolates in CRL11171 cells maintained in a medium supplemented with FBS was compared with virus propagation in the same cell line maintained in the same medium with a serum replacement. The PRRSV replicated significantly better when the cell culture medium was supplemented with FBS. The results of this study have implications for the use of serum replacement-supplemented medium for PRRSV diagnosis by virus isolation. PMID- 11482615 TI - Active counterpositioning or orthotic device to treat positional plagiocephaly? AB - Active counterpositioning and orthotic helmets are the two main nonsurgical management options for positional plagiocephaly. The purpose of this study was to compare these two management regimens. We included a random sample of infants referred between January 1, 1998 and October 31, 1999 to Middlemore Hospital and Auckland Surgical Center, for management of positional plagiocephaly. Two dimensional head tracings were taken for each infant, every 3 to 12 months. From these tracings, we obtained Cranial Index and Cranial Vault Asymmetry Index. Seventy-nine infants were assessed during an average of 48.2 weeks. Five infants had normal head tracings, and were therefore excluded from the study. Of the 74 infants included in this study, 45 were managed with active counterpositioning, and 29 with orthotic helmets. Average management time for active counterpositioning was 63.7 weeks, and 21.9 weeks for orthotic helmet treatment. For infants managed with active counterpositioning, the average change in Cranial Vault Asymmetry Index was 1.9%. In the orthotic group, average change in Cranial Vault Asymmetry Index was 1.8%. Orthotic helmets have an outcome comparable to that of active counterpositioning, although the management period is approximately three times shorter. Active counterpositioning generally had a slightly better outcome than orthotic management after the management period. PMID- 11482616 TI - Reduction mandibuloplasty: ostectomy of the lateral cortex around the mandibular angle. AB - Many surgical techniques have been applied toward correcting the prominent mandibular angle. One of the most popular methods is mandibular angle ostectomy. However, gross asymmetry, mandibular condylar fracture, and unnatural contouring of the mandibular angle are frequent consequences. Therefore, we perform ostectomy of the lateral cortex around the mandibular angle in patients with wide and squared lower faces. Seventeen female patients with ages ranging from 23 to 35 years underwent surgery. Ostectomy of the lateral cortex of the mandible and mandible angle ostectomy were performed in five patients (group I). Ostectomy of the lateral cortex only was performed in the remaining 12 patients (group II). The follow-up period ranged from 1 to 5 years. Postoperatively, the bigonial distance was effectively reduced in both groups (group I, mean 13 mm; group II, mean 12 mm). Compared with group I, the gonial angle and mandibular plane-sella nasion angle were within normal ranges in group II. We therefore conclude that the ostectomy of the lateral cortex alone allows the reduction to be achieved without changing the angle of the mandible itself. Moreover, although the resection of the mandibular angle was not performed, the contour of mandibular angle became soft, giving a more natural appearance. PMID- 11482617 TI - Reliable, minimally invasive oromandibular reconstruction using metal plate rolled with pectoralis major myocutaneous flap. AB - The purpose of this study was to minimize the surgical invasiveness to the donor site and the amount of the primary reconstruction time after oromandibular tumor resection. Oromandibular reconstruction was performed only using a pectoralis major myocutaneous flap and a metal plate. The pectoralis major myocutaneous flap was grafted to the oral cavity defect by rolling and wrapping around the metal plate with the muscle of the flap. No early postoperative complications have been noted in all seven patients. An average of 2 years and 1 month has past since surgery, and to date no infections, plate exposure, or plate breakage have been observed in any of the patients. The safety of the oromandibular reconstruction using a metal plate was improved by rolling the muscle of the pectoralis major myocutaneous flap around the metal plate. The present method was shown to be a rational technique that allowed primary reconstruction of the oral cavity and mandible in a minimally invasive manner in a short time. PMID- 11482618 TI - Early treatment of severe mandibular hypoplasia with distraction mesenchymogenesis and bilateral free fibula flaps. AB - The technique of distraction has revolutionized the treatment of mandibular hypoplasia; however, presently large mandibular defects still require bone grafts. Microvascular grafting is commonly used in adults. Conversely, in pediatric reconstruction, nonvascularized rib grafts remain standard. Unfortunately, resorption of nonvascularized bone remains a major issue, particularly when soft tissue is hypoplastic. This case study represents a combination of techniques in the treatment of severe mandibular deficiency, and introduces the concept of distraction mesenchymogenesis. The patient was a 2 1/2 year-old boy with severe bilateral Pruzansky class III mandibular hypoplasia. He had a permanent open mouth posture, an overjet of 23 mm, and was unable to move the lower mandibular segment. His oropharyngeal airway diameter was 2.2 mm and he was tracheostomy dependent. The patient was treated with distraction of the lower jaw mesenchyme followed by bilateral functional free fibular microvascular flaps containing reinnervated muscle. This created a well-vascularized body, ramus, and condyle bilaterally within an adequate soft-tissue envelope. Postoperatively, the overjet was reduced to 5 mm. The patient can now actively move his mandible. Airway diameter increased to 10 mm, and the patient is able to tolerate intermittent tracheostomy plugging. This innovative combination of techniques allows early intervention, limits graft resorption, and improves airway control. PMID- 11482619 TI - Melanotic neuroectodermal tumor of infancy involving the mandible: 7-year follow up after hemimandibulectomy and costochondral graft reconstruction. AB - Oral and maxillofacial tumors occur rarely in the pediatric population compared with the adult population. We report a case of a 6-months old female infant suffering from a melanotic neuroectodermal tumor of infancy involving the mandible. Tumor resection was performed using a submandibular approach; the mandibular defect was reconstructed primarily with autogenous costochondral grafts. During a 7-year follow-up period, there has been no tumor recurrence. The costochondral graft healed well; tracing of panoramic radiographs at 2, 3, and 6 years documented some vertical overgrowth and growth retardation in the transversal dimension. The authors conclude that the use of costochondral grafts despite its controversial role for mandibular reconstruction can be recommended in particular after continuity resections in newborn infants. However, long-term follow-up is necessary as well as secondary corrective surgery at early skeletal maturity. PMID- 11482620 TI - Rare craniofacial anomaly: Tessier no. 2 cleft. AB - Four cases of facial cleft that fit the anatomic description of the rare Tessier no. 2 cleft, with two patients having the no. 12 cleft extending to the cranium as no. 2 clefts, are presented. In all patients, clinical expressions of the anomaly were different. Thus, diverse surgical procedures were used in all cases. These cases and review of the literature help to define the soft-tissue and bony course of these clefts, and also emphasize the role of three-dimensional computed tomography scan imaging to show the bony cleft route. The diagnosis and treatment plan of the no. 2 cleft as well as its cranial counterpart are discussed in this report. PMID- 11482621 TI - Postoperative infections in craniofacial reconstructive procedures. AB - The rate of, and possible risk factors for, postoperative craniofacial infection is unclear. To investigate this problem, we reviewed 349 cases of craniofacial skeletal procedures performed from 1996 to 1999 at our institution. Infection rate was determined and correlated with the use of implants, operative site, and cause of deformity. The inclusion criteria consisted of all procedures requiring autologous or prosthetic implantation in craniofacial skeletal sites, as well as all procedures involving bone or cartilage resection, osteotomies, debridement, reduction and/or fixation. Procedures that did not involve bone or cartilage surgery were excluded. The criteria for diagnosis of infection included clinical confirmation and one or more of 1) intravenous or oral antibiotic treatment outside of the prophylactic surgical regimen; 2) surgical intervention for drainage, irrigation, and or debridement; and 3) microbiological confirmation. Among the 280 surgical cases that fit the inclusion criteria and had complete records, there were 23 cases of postoperative infection (8.2%). The most common site for postoperative infection was the mandible (infection rate = 16.7%). Multiple logistic regression analysis revealed gunshot wound to be the most significant predictor of postoperative infection. Additionally, porous polyethylene implantation through a transoral route was correlated with a significant risk of postoperative infection. PMID- 11482622 TI - Free fibular flap reconstruction in mandibular osteopetrosis. AB - Extensive involvement of the mandible with sclerosis secondary to the rare genetic disorder, osteopetrosis, is a difficult reconstructive problem. It is typically refractory to conservative management or local surgical methods because of vascular compromise of the bone. For this reason, total resection of all involved bone with microvascular graft reconstruction is an ideal method for mandibular reconstruction in osteopetrosis that has not been previously reported. PMID- 11482623 TI - Surface anatomy of the face in Down's syndrome: linear and angular measurements in the craniofacial regions. AB - Measurements (23 projective linear, 2 angular) taken in the 6 craniofacial regions of 127 patients with Down's syndrome showed that 63.1% (1,836 of 2,908) were within normal limits and 36.9% (1,072) were outside them. Abnormal measurements were subnormal in 90.8% (973) and supernormal in 9.2% (99). All statistical summaries were based on z scores (adjusting for age and sex differences) classified into a small number of ranges to yield a simplified frequency distribution for each measurement. The purpose of the study was to identify the measurements closest to normal and those indicating the most severe degrees of sub- or supernormality. Approximately a quarter of normal measurements were classified as optimal, and half the subnormal or supernormal measurements were classified as severe. Intercanthal width had the highest frequency of optimal measurements (93.7%, 119 of 127), head circumference the smallest (28.6%, 36 of 126). Knowledge of the frequency of extreme abnormalities in the craniofacial regions will help during visual examination of patients with Down's syndrome. This study found the highest percentage of severely subnormal measurements in the orbital region (57.8%, 74 of 128) and the smallest in the labio-oral region (32.7%, 16 of 49). The measurement with the highest proportion of severely subnormal to all subnormal values was the palpebral fissure length (68%, 51 of 75), and the nose width had the smallest proportion (14.3%, 1 of 7). PMID- 11482624 TI - Deep middle masseteric artery (dMMA) attributed to hemorrhage in resection of masseter muscle and mandibular angle. AB - This study aimed to investigate pertinent arterial supplies of masseter muscle to prevent fatal hemorrhage in resection of masseter muscle and/or mandibular angle ostectomy. Fifty-three postmortem cadavers of Koreans were used for the work. Color latex was injected into the arteries to outline 17 of 53 specimens. We found that an artery branches off the external carotid artery and enters the masseter muscle at the midpoint of its posterior margin 31 mm above the gonion. We termed this the middle masseteric artery. The middle masseteric artery is divided into superficial and deep branches. The deep branch of the middle masseteric artery travels deep in the muscle close the periosteum of the mandible in 94% of cases. The average diameter is 1.23 +/- 0.26 mm. A small artery with 1.23-mm diameter is enough to cause massive bleeding if severed. The deep branch of the middle masseteric artery is vulnerable in such procedures as resection of the masseter muscle and/or ramus and angle of the mandible. PMID- 11482625 TI - TGF-beta isoform and receptor expression during scarless wound repair. PMID- 11482626 TI - Role of TGF-beta signaling in the regulation of programmed cranial suture fusion. PMID- 11482627 TI - Extrinsic tension results in FGF-2 release, membrane permeability change, and intracellular Ca++ increase in immature cranial sutures. AB - There are numerous studies cataloging the temporal profiles of the various growth factors during the morphogenesis of cranial sutures. There are also many clearly documented mutations of the receptors of some of these growth factors such as fibroblast growth factor (FGF)R-2 and FGFR-3 in clinical craniosynostosis. It is obvious, and often concluded, that growth factors play a role or are involved in craniofacial development. However, precisely what that role is, what causes the changes in the growth factor levels, and why these changes occur in the particular temporal and spatial patterns observed remains elusive. Using simple physics, we applied a plasma membrane disruption model and the principles of complex adaptive systems to arrive at a conjecture of calvarial morphogenesis. The purpose of this article is to introduce the concept of complex adaptive systems, to propose our conjecture, and to provide experimental proof of some key steps in this conjecture: tension induces rapid and demonstrable physiological responses in some cells within the immature cranial sutures. These responses include increases of intracellular Ca++, plasma membrane permeability, and the release of growth factors, e.g., FGF-2. Paired coronal sutures from 1-week-old Sprague-Dawley rat pups were subjected to either 0.59 N of tensile force or no force for 5 minutes in a protein-free medium. FGF-2 levels in the media were measured by slot blot analysis. Western blot analysis was used to determine FGF-2 levels in the sutures. To determine cell membrane permeability changes, fluorescein-conjugated dextran, with a molecular weight of 10 kd, was added to the media during the 5 minutes with or without tensile force. Laser confocal microscopy was used to compare the amount of entry of this impermeant tracer and the pattern of permeability change at the tissue level. To determine the intracellular pCa++, the sutures were first loaded with a calcium indictor, FURA 2 AM, and then subjected isotonically to 0.059 N of tension. The intracellular pCa++ was expressed as ratio of Ca++-bound FURA-2 to Ca++-free FURA-2. The experimental findings were as follows: 1) Sutures, in response to tension, release FGF-2. 2) Sutures contain higher levels of FGF-2 when strained. 3) There is an increase in the sutural cell membrane permeability as a result of tensile strain. 4) The cells along the leading edges of the ossification fronts (at the insertion sites of Sharpey's fibers) demonstrated the maximum permeability increase. 5) There was an immediate (within seconds) increase in intracellular Ca++. and 6) This increase in intracellular Ca++ caused by tension was reversible and independent of the extracellular Ca++ ion availability. In summary, these data support, in part, the conjecture that growth of the brain places strain on the cells within the immature sutures, which causes the iteration of a set of cellular subroutines. These subroutines integrate to generate the emergent property of directed cranial expansion with dissipation of the initiating strains. PMID- 11482628 TI - FGF-4 or FGF-2 administration induces apoptosis, collagen type I expression, and mineralization in the developing coronal suture. PMID- 11482629 TI - Quantitative assessment of cranial defect healing and correlation with the expression of TGF-beta. AB - Circular parietal defects from 3 to 12 mm in diameter were made in 45 6-month old skeletally mature guinea pigs, and animals were sacrificed after survival periods of 3 days to 12 weeks. The original defect was harvested in continuity with a rim of surrounding bone and the adjacent dura and pericranium. After 12 weeks, all 3 and 5 mm defects were completely covered by a bridge of bone, while residual defects were noted within the 8 and 12 mm wounds. Percentage of new bone formation was significantly higher within 3 mm defects, than in all larger defects at each time interval from 1 week on (P < .05), reaching a mean of 93% in 3 mm defects and remaining below a mean of 31% in the remaining defect sizes. Immunolocalization demonstrated an osteogenic front in which the osteoblasts stained strongly for all isoforms of TGF-beta, with the intensity decreasing after the majority of the defects had reossified; this front was located at the advancing bone edge of the defect as well as the endocranial side adjacent to the dura. In conclusion, isoforms of TGF-beta are upregulated during a limited "window" of time corresponding to the period of calvarial reossification, and are localized to osteoblasts within an osteogenic front at the periphery and dural surfaces of the defects. PMID- 11482630 TI - Psychosocial work characteristics and perceived control in relation to cardiovascular rewind at night. AB - This study examined the effects of psychosocial work characteristics on cardiovascular rewind at night. Ambulatory 24-hr recordings of blood pressure (BP) and heart rate (HR) of 75 borderline hypertensive and 74 normotensive men were related to diary ratings of perceived control (PC) and to scores of psychological demand (P), control (C), and social support (S) at work determined by an occupational classification system. Multiplicative interaction terms for job strain (P x C), isostrain (P x C x S), and Job Strain x Perceived Control (P x C x PC) were calculated. The P x C x PC interaction predicted diastolic BP at night but not at work. A delayed latency to attain the lowest systolic BP during the night was found for jobs with high job strain and isostrain. Low perceived control and social support were associated with higher HR at work and at night. A logistic regression analysis indicated that the interaction between P x C x PC and the body mass index was independently associated with borderline hypertension. PMID- 11482631 TI - The interactive effect of chronic exposure to noise and job complexity on changes in blood pressure and job satisfaction: a longitudinal study of industrial employees. AB - The hypothesis of this study was that noise exposure level and job complexity interact to affect changes in blood pressure (BP) levels and job satisfaction over 2-4 years of follow-up. Results showed that among workers exposed to high noise, those with complex jobs showed increases in BP that were more than double shown by those with simple jobs. Under low noise exposure, there was a small increase in BP for workers with complex jobs but about a 3-fold increase in workers with simple jobs. The prevalence of elevated BP showed a similar trend. Job satisfaction increased among workers with complex jobs but was much less in those exposed to high noise. It was concluded that exposure to occupational noise has a greater negative impact on changes in BP and job satisfaction over time among those performing complex jobs. In contrast, job complexity had a clear beneficial effect for workers exposed to low noise. PMID- 11482632 TI - Work, recovery activities, and individual well-being: a diary study. AB - This study extends previous research on respite from work and addresses the question of how individuals use their leisure time to recover from work. It is hypothesized that time spent on work-related and household activities has a negative effect on well-being, whereas low-effort, social, and physical activities are assumed to have a positive effect. One hundred Dutch teachers completed a diary on leisure time activities and situational well-being for 5 days, and work situation variables were assessed with a questionnaire. Multilevel analyses in which preleisure well-being and work situation variables were entered as control variables supported 4 of the 5 hypotheses. Moreover, a lagged effect of high time pressure on poor situational well-being was found. The study showed that leisure time activities and a low-stress work situation contribute independently to an individual's well-being. PMID- 11482633 TI - Designing a safer workplace: importance of job autonomy, communication quality, and supportive supervisors. AB - The direct and indirect effects of work characteristics on self-reported safe working were investigated in a longitudinal study of frontline manufacturing employees (N = 161). The work characteristics included job autonomy, role overload, role conflict, supportive supervision, training adequacy, job security, and communication quality. Job autonomy and communication quality were positively associated with safe working after prior levels of these variables were controlled for, and supportive supervision had a lagged positive effect on safe working 18 months later. Additional analyses showed that organizational commitment fully mediated the effect of job autonomy on safe working and partially mediated the effect of communication quality on safe working. The study suggests that work characteristics are important antecedents of safe working. PMID- 11482634 TI - Correlates of work injury frequency and duration among firefighters. AB - This study examined demographic, personality, and economic incentive correlates of workplace injuries suffered by 171 firefighters over a 12-year period. Results showed that female firefighters experienced more injuries than male firefighters. Several Minnesota Multiphasic Personality Inventory (MMPI) scales (Conversion Hysteria, Psychopathic Deviate, and Social Introversion) were positively related to injury frequency. Regression analyses revealed that age, tenure, gender, marital status, type of injury, and wage variables were significant when predicting the duration of injuries as well as an interaction between marital status and gender. Two MMPI scales (Psychopathic Deviate and Schizophrenia) were also significantly related to injury duration. Indemnity cost estimates were calculated. The results underscore the importance of distinguishing the duration of injury from the occurrence of injury. PMID- 11482635 TI - Workplace substance abuse prevention and help seeking: comparing team-oriented and informational training. AB - Employees fail to seek help for alcohol or drug (AOD) abuse because of unhealthy work climates, stigma, and distrust in Employee Assistance Programs (EAPs). To address such problems, the authors randomly assigned groups of municipal employees (N = 260) to 2 types of training: a 4-hr informational review of EAPs and policy and an 8-hr training that embedded messages about AOD reduction in the context of team building and stress management. Pre- and posttraining and 6-month follow-up surveys assessed change. Group privacy regulation, EAP trust, help seeking, and peer encouragement increased for team training. Stigma of substance users decreased for information training. EAP/policy knowledge increased for both groups. A control group showed little change. Help seeking and peer encouragement also predicted EAP utilization. Integrating both team and informational training may be the most effective for improving help seeking and EAP utilization. PMID- 11482636 TI - Behind closed doors: in-home workers' experience of sexual harassment and workplace violence. AB - The authors developed and tested a structural model predicting personal and organizational consequences of workplace violence and sexual harassment for health care professionals who work inside their client's home. The model suggests that workplace violence and sexual harassment predict fear of their recurrence in the workplace, which in turn predicts negative mood (anxiety and anger) and perceptions of injustice. In turn, fear, negative mood, and perceived injustice predict lower affective commitment and enhanced withdrawal intentions, poor interpersonal job performance, greater neglect, and cognitive difficulties. The results supported the model and showed that the associations of workplace violence and sexual harassment with organizational and personal outcomes are indirect, mediated by fear and negative mood. Conceptual implications for understanding sexual harassment and workplace violence, and future research directions, are suggested. PMID- 11482637 TI - Influence of solution composition and column aging on the reduction of nitroaromatic compounds by zero-valent iron. AB - Granular iron is used in reactive permeable barriers for the reductive treatment of organic and inorganic groundwater contaminants. The technology is well established, however, its long-term performance and the importance of the groundwater composition are not yet well understood. Here, the influence of chloride, nitrate, silicate, and Aldrich humic acid on the reactivity of Master Builder iron was studied under anoxic conditions using small packed columns and 2 nitrotoluene (2-NT) as a model contaminant. After initially complete reduction of 2-NT to 2-aminotoluene (2-AT) in the column, possibly under mass-transfer controlled conditions, the reactivity of the iron was found to decrease substantially. In the presence of chloride, this decrease was slowed while exposure to silicate resulted in a very quick loss of iron reactivity. Nitrate was found to interfere strongly with the effect of chloride. These observations are interpreted in terms of corrosion inhibition/promotion and competition. Our results suggest that reactive barrier performance may be strongly affected by the composition of the treated groundwater. PMID- 11482638 TI - Influence of n-alkanes and petroleum on fatty acid composition of a hydrocarbonoclastic bacterium: Marinobacter hydrocarbonoclasticus strain 617. AB - This study concerns the effects of various long-chain n-alkanes, n-alkane mixtures and Arabian Light crude oil on the fatty acid (FA) composition of a sedimentary marine bacteria (Marinobacter hydrocarbonoclasticus strain 617), growing under aerobic conditions. The cultures with n-alkanes, as compared with soluble carbon sources, led to greater amounts of saturated and methyl branched FA (mainly belonging to a delta10 series). We observed the appearance or increase of saturated and unsaturated FA with the same carbon chain length (CCL) as the n alkane carbon source (maximum for n-alkane CCL corresponding to the 'range' of the de novo synthesized fatty acids). We also observed a strong control of the oddness/evenness of the CCL of the FA by the oddness/evenness of the n-alkane. A n-alkane utilization index, (saturated + branched)/monounsaturated fatty acids (SAFA + BFA/MUFA) enabled discriminating between soluble carbon sources and hydrocarbons. PMID- 11482639 TI - Preparation and certification of a reference material on PCBs in pig fat and its application in quality control in monitoring laboratories during the Belgian "PCB crisis". AB - In this article, the production and validation of a new certified reference material "PCBs in animal fat" for the control of the maximum level of 200 ng/g setup by the European Communities for veterinary products from Belgium is described. Three materials are established: a blank, one material with about 100 ng/g and one with about 200 ng/g (sum of seven PCBs). Data on the production and certification are given. Additionally, this material was used as an unknown test material in the quality assurance program of the Belgium meat monitoring system (before the certification of the material). While the certification was performed with an uncertainty of less than 10%, the round robin exhibited larger deviations. However, these deviations were less than 20% for most of the 30 participating laboratories. Only two had significantly higher deviations. PMID- 11482640 TI - A rapid and sensitive method for methyl tert-butyl ether analysis in water samples by use of solid phase microextraction and gas chromatography-mass spectrometry. AB - This work describes a rapid and sensitive solid-phase microextraction (SPME) method for the isolation and analysis of methyl tert-butyl ether in water samples. Methyl tert-butyl ether was extracted from aqueous solutions using SPME fibre coated with Divinylbenzene/Carboxen/polydimethylsiloxane (30 microm film thickness) and analysed by GC-MS with a Hewlett Packard 6890/5973 system equipped with a capillary column coated with Vocol (30 m x 0.25 mm, 1.5 microm film thickness). Extraction parameters and chromatographic separation conditions were optimised. The developed method showed good analytical performance in terms of precision (RSD between 2% and 8%) and accuracy (mean recovery from 96% to 104%) with a detection limit of 14 ppt. Finally the method was applied to surface, tap and commercial mineral water samples, as well as snow samples collected along a busy road of Bologna town area. The median concentration of methyl tert-butyl ether in all these samples (0.05-0.4 ppb) was well below the maximum aqueous contamination levels in water adopted in the United States (13 ppb). PMID- 11482641 TI - Effects of fulvic acid on the bioavailability of rare earth elements and GOT enzyme activity in wheat (Triticum aestivum). AB - Fulvic acid (FA) was extracted and purified from natural soil and the effects of such FA on the bioaccumulation of rare earth elements (REEs, La3+, Gd3+ and Y3+) in wheat seedling were investigated. The results indicated that low concentration of FA (<0.4 mg C/l to root, <0.7 mg C/l to tops (stem and leaves)) could increase the bioaccumulation values of REEs in wheat, but when the concentration of FA was high (>0.4 mg C/l to root, >1.5 mg C/l to tops) the bioaccumulation values were decreased. Kinetic experimental results suggested that bioaccumulation values of REEs in roots for 30 days were correlated with the kinetic linear growth equation, and correlation coefficients were higher than 0.861. The kinetic bioaccumulation pattern of REEs in tops was different from that in root. The bioaccumulation values of REEs in wheat root were much higher than in wheat tops. Variations of glutamic oxaloacetic transaminase (GOT) enzyme activities in wheat root and tops were determined. A good correlation existed between the bioaccumulation values of REEs and GOT enzyme activities, and the correlation coefficients were higher than 0.922. GOT is an important parameter influencing the bioavailability of REEs. PMID- 11482642 TI - Modification of the Zahn-Wellens test: determination of the biodegradability of poorly soluble, adsorbing and volatile substances by measurement of oxygen consumption and carbon dioxide production. AB - Based on the Zahn-Wellens test (OECD 302 B, 1992; DIN EN ISO 9888, 1999), a test system has been developed which enables a continuous and parallel determination of oxygen consumption (pressure measurement) and carbon dioxide production (conductivity measurement). It is a closed test system consisting of a culture flask, a carbon dioxide absorption flask, a pump as well as integrated measuring and control instruments. The air circulating within the test system causes the carbon dioxide present in the test solution to be stripped out completely and directly absorbed by the absorption solution. Avoiding costly thermostatting of the test apparatus, the results of the pressure measurements were temperature corrected arithmetically. The functional reliability of the measuring apparatus has been demonstrated exemplary in degradation experiments with selected test substances. This new test system also facilitates to test poorly soluble, adsorbing and volatile substances for inherent biodegradability and constitutes an appropriate complement to the standardised Zahn-Wellens test. The Federal Environmental Agency will use it as input to international standardisation activities ongoing within the OECD, as a draft standard. PMID- 11482643 TI - A critical compilation of Henry's law constant temperature dependence relations for organic compounds in dilute aqueous solutions. AB - A comprehensive compilation of published studies reporting directly measured experimental determinations of Henry's law constant (HLC) temperature dependence relationships for organic compounds in dilute, non-saline aqueous solutions under ambient conditions was conducted. From this effort, 55 such studies (covering 204 organic compounds) were identified, critically reviewed, summarized and discussed. Of the 204 compounds, 57 were studied in more than one investigation. For the 57 'multi-studied' compounds, relatively good agreement (averaging within 20-30%) was found between the results from different investigations. Given such results, a 'consensus' relationship (i.e., an average temperature dependence relation) was generated for each of the multi-studied compounds. Overall, considering relations established for 197 of the 204 compounds investigated (the results for the other 7 compounds being excluded due to poor correlation coefficients), the average slope of the temperature dependence line was found to correspond to an increase in HLC values by a factor of 1.88 (i.e., an 88% increase) per 10 degrees C rise in temperature (thermodynamically equivalent to an average enthalpy of volatilization of 47 kJ/mole). The associated range found for the temperature dependence slope corresponds to an increase in HLC values by a factor ranging from 1.12 to 3.55 (i.e., a 12-255% increase) per 10 degrees C rise (equivalent enthalpy of volatilization range: 8-93 kJ/mole). The wide range of slope values found indicates that serious errors may result if one applies the commonly cited 'rule of thumb' that HLC values double per 10 degrees C rise in temperature to a specific compound. In light of this finding, when faced with a lack of data, a prudent course for practitioners to take appears to be conducting a laboratory study to determine the exact temperature dependence for the compound(s) of interest. PMID- 11482644 TI - Microbial degradation of mefenoxam in rhizosphere of Zinnia angustifolia. AB - The fate of the fungicide mefenoxam was studied in a containerized rhizosphere system. The rhizosphere system used Zinnia angustifolia (Tropic Snow) in a bark/sand potting mix and was compared to bulk potting mix (no plants). Rhizosphere microbial populations were allowed to establish for 3 weeks prior to fungicide addition (20 microg per g mix). Mefenoxam and degradation product concentrations were determined by High HPLC or capillary electrophoresis after extraction. Seventy eight percent of the fungicide originally applied to the rhizosphere was degraded after 21 days compared to 44% in bulk system (no plant). The primary degradation product was the free acid N-(2,6-dimethylphenyl)-N (methoxyacetyl)-DL-alanine, which accounted for 71% of the applied parent chemical after 30 days. N-(2,6-dimethylphenyl)-acetamide was also detected, but in lesser amounts. Bacterial populations in the rhizosphere increased during the 30-day period, which correlated with an increase in degradation of the parent compound. Pure cultures of Pseudomonas fluorescens and Chrysobacterium indologenes isolated from the rhizosphere system could degrade the applied fungicide (10 microg/ml) almost completely to the free acid within 54 h. PMID- 11482645 TI - Short-term exposure to sub-lethal doses of lindane affects developmental parameters in Chironomus riparius Meigen, but has no effect on larval glutathione S-transferase activity. AB - Chironomus riparius Meigen were exposed to 0, 0.01, 0.1, 0.5, 0.75 and 1.0 ppm lindane for 48 h as fourth instar larvae. Exposure had no effect on glutathione-S transferase (GST) activity in larvae snap-frozen immediately following exposure. In contrast, exposure had longer-term consequences affecting developmental parameters. Concentrations above 0.5 ppm lindane affected larval behaviour, reduced adult body size and fecundity and delayed emergence times. The lack of significant change in GST activity when life history characters were affected by high concentrations of lindane, suggests that in C. riparius, GST is not a sensitive biomarker of pesticide exposure or effect. PMID- 11482646 TI - Seasonal dependence of the chiral composition of alpha-HCH in coastal deposition at the North Sea. AB - The environmental pollutant alpha-HCH was analysed in seawater, air and bulk deposition samples from the North Sea. Enantioselective analyses of the sample extracts gave evidence of the change of the direction of net air-water gas transfer of the contaminant in dependence on the season. Due to warmer water surface temperatures during late summer to early autumn the equilibrium of alpha HCH between air and water is dominated rather by volatilisation than deposition. The volatilisation of non-racemic alpha-HCH, as known to occur from seawater, changes the enantiomeric ratio in air, which is reflected in the observed ratio in rain that passes the air column. PMID- 11482647 TI - Evaluation and comparison of multimedia mass balance models of chemical fate: application of EUSES and ChemCAN to 68 chemicals in Japan. AB - The European Union System for Evaluation of Substances (EUSES) and the ChemCAN chemical fate model are applied to describe the fate of 68 chemicals on two spatial scales in Japan. Emission information on the chemicals has been obtained from Japan's Pollutant Release and Transfer Registry and available monitoring data gathered from government reports. Environmental concentrations calculated by the two models for the four primary environmental media of air, water, soil and sediment agree within a factor of 3 for over 70% of the data, and within a factor of 10 for over 87% of the data. Reasons for certain large discrepancies are discussed. Concentrations calculated by the models are generally consistent with the lower range of concentrations that are observed in the environment. Agreement between modeled and observed concentrations is considerably improved by including an estimate of the advective input of chemicals in air from outside Japan. The agreement between the EUSES and ChemCAN models suggests that results of individual chemical assessments are not likely to be significantly affected by the choice of chemical fate model. Primary sources of discrepancy between modeled and observed concentrations are believed to be uncertainties in emission rates, degradation half-lives, and the lack of data on advective inflow of contaminants in air. PMID- 11482648 TI - Abiotic Fe(III) induced mineralization of phenolic substances. AB - The redox process between iron(III) (in dissolved form and as the mineral phase ferrihydrite) and phenolic substances has been examined. We investigated the relationship between the structure and reactivity for the dihydrobenzene reductants catechol, hydroquinone and resorcine, and for the 2-methoxyphenol guaiacol with iron(III), by determining the rate of the Fe(III) reduction as well as the production of CO2. This work demonstrates that catechol and guaiacol will be effectively oxidized to CO2 by reducing iron(III). Hydroquinone shows a reduction of iron(III), but no accompanying mineralization could be determined. In contrast, resorcine showed no reaction with Fe(II). The deciding factor on whether or not mineralization occurs were controlled by the position of the hydroxy groups. It is shown that phenolic substances with two hydroxy groups in the orthoposition or at least one hydroxy group and a methoxy group can be oxidized to CO2 while iron(III) is reduced. PMID- 11482649 TI - Effect of photosensitizer riboflavin on the fate of 2,4,6-trinitrotoluene in a freshwater environment. AB - The effect of riboflavin (1 microM) on the fate of TNT (20 mg/l) in a natural water environment was studied. The relative contribution of photolysis, microbial assemblages and freshwater matrix to TNT degradation was examined. The rates, extent and products of TNT and riboflavin transformation were compared under different experimental conditions. It was found that riboflavin significantly enhanced the degradation of TNT in natural water environment. Thus it is a potentially useful photosensitizing agent for the treatment of TNT-contaminated surface water. Furthermore, in the presence of riboflavin, two new intermediates with max. absorption wavelength of 230 nm were found, demonstrating that transformation of TNT in the presence of riboflavin undergoes different pathways. PMID- 11482650 TI - Role of pH in partitioning and cation exchange of aromatic amines on water saturated soils. AB - Predicting the reversible interactions between aromatic amines and soil is essential for assessing the mobility, bioavailability and exposure from contaminated sites. Reversible sorption mechanisms of aniline and alpha naphthylamine were investigated by using single and binary solute sorption to five soils at several pH values, and by applying a distributed parameter (DP) model. The DP model assumes linear partitioning of the neutral species into soil organic matter domains and organic cation binding on negative-charged sites with the exchange coefficients represented by a Gaussian probability distribution. Sorption nonlinearity was attributed to cation exchange with varying site affinities, which was adequately simulated using the DP model. Greater uptake by hydrophobic partitioning and selectivity for cation exchange sites was observed for alpha-naphthylamine compared to aniline. Sorption of alpha-naphthylamine was not impacted quantitatively by aniline under those conditions examined; however, aniline sorption was reduced by alpha-naphthylamine with the largest reduction occurring in the soil with the lowest pH. DP model simulations showed that although hydrophobic partitioning increases with soil-solution pH, cation exchange still contributes significantly to the total sorption even at soil solution pH values greater than pKa + 2. PMID- 11482651 TI - On the leachability and sources of some elements in sediments from the Bothnian Sea and the Gotland Deep (the Baltic Sea). AB - The partitioning of some elements in sediments of two Baltic Sea basins (the Bothnian Sea and the Gotland Deep) was established by leaching experiments. Both partial (dithionite-citrate-bicarbonate) and "total" (HCl-KClO3) leach were used. Trace metals of both natural and anthropogenic sources (Co, Cu, Ni, Pb, Zn) were evaluated for affinity to terrestrial material and to authigenic mineral elements (Fe, S). The results indicate that the easily leachable "mobile" fraction is not necessarily a good indicator for pollution. Longer stagnation periods result in intensive pyrite formation in the Gotland Deep which mask pollution trends. The permanent oxic conditions in the Bothnian Sea may affect a dissolution capability for selected elements. PMID- 11482652 TI - Strategies for including vegetation compartments in multimedia models. AB - The incentives for including vegetation compartments in multimedia Level I, II and III fugacity calculations are discussed and equations and parameters for undertaking the calculations suggested. Model outputs with and without vegetation compartments are compared for 12 non-ionic organic chemicals with a wide variety of physical-chemical properties. Inclusion of vegetation compartments is shown to have a significant effect on two classes of chemicals: (1) those that are taken up by atmospheric deposition and (2) those that are taken up by transpiration through the plant roots. It is suggested that uptake from the atmosphere is important for chemicals with logK(OA) greater than 6 and a logK(AW) of greater than -6. Plant uptake by transpiration is important for chemicals with logK(OW) less than 2.5 and a logK(AW) of less than -1. At logK(OA) > 9 atmospheric uptake is dominated by particle-bound deposition and the importance of partitioning to vegetation is largely dependent on the relative magnitude of the particle deposition velocities to soil and vegetation. These property ranges can be used to determine if a chemical will significantly partition to vegetation. If the chemical falls outside the property ranges of the two classes it will probably be unnecessary to include vegetation in models for assessing environmental fate. The amount of chemical predicted to partition to vegetation compartments in the model is shown to be highly sensitive to certain model assumptions. Further experimental research is recommended to obtain more reliable equations describing equilibrium partitioning and uptake/depuration kinetics. PMID- 11482653 TI - Effects of redox potential and pH value on the release of rare earth elements from soil. AB - Equilibrium release experiments were conducted under three different pH values of 3.5, 5.5 and 7.5 as well as three redox potentials of 400, 0 and -100 Mv to investigate the influence of redox potential and pH value on the La, Ce, Gd and Y release of from the simulated-REEs-accumulation (SRA) soil. Oxygen and nitrogen were allowed to flow into soil suspension to adjust redox potential to a preset value, and 1 mol/l HCl or 1 mol/l NaOH solutions were added into the soil suspension to keep pH at a preset value. Results indicated that La, Ce, Gd and Y release increased gradually with the decrease of pH value or Eh, and the influence of redox potential on Ce was more remarkable than on La, Gd and Y. At the same time. It was observed that La, Ce, Gd and Y releases were positively correlated with the release of Fe and Mn, indicating that La, Ce, Gd and Y releases might originate from dissolution of Fe-Mn oxyhydroxides under reduction and low pH conditions. Moreover, it was found that alteration of pH value and redox potential might affect the change of La, Ce, Gd and Y species in the soil. The contents of La, Ce, Gd and Y in exchangeable fraction and Fe-Mn oxide fraction in the solid phase from soil suspension separation decreased with the decline of pH value and redox potential. Multiple stepwise regression analysis showed that exchangeable fraction and Fe-Mn oxide fraction predominately contributed to the La, Ce, Gd and Y release. Low pH value and redox potential were more favorable to La, Ce, Gd and Y releases following the change of their species. The La, Ce, Gd and Y contents in exchangeable fraction and Fe-Mn oxide fraction are the main contributors to their release. PMID- 11482654 TI - Study on the kinetics of cerium(III) adsorption-desorption on different soils of China. AB - The kinetics of Ce(III) adsorption-desorption on four typical soils in China has been studied by using the batch method with the radioactive nuclide 141Ce. Results indicated that Ce(III) adsorption was rapid and nearly finished in less than 0.5 min. Desorption procedure was about completed in 1-30 min in the tested soils. Ce(III) desorption equilibrium times vary with different soils. The amounts of Ce(III) desorption on different soils in the same time were different. The Elovich equation proved to be the best models for fitting the data of Ce(III) desorption reactions in fluvo-aquic soil and black soil; and the parabolic diffusion equation was the best model in red earth and loess soil. PMID- 11482655 TI - A new potential toxaphene congener: synthesis, GC/EI-MS study, crystal structure, NMR analysis, and ab initio calculations of 3-endo,5-endo-dichloro-7,7-bis chloromethyl-4-dichloromethyl-tricyclo[2.2.1.0(2,6)]heptane. AB - A new potential toxaphene congener 3-endo,5-endo-dichloro-7,7-bis-chloromethyl-4 dichloromethyl-tricyclo[2.2.1.0(2,6)]heptane 2 has been isolated from reaction mixture obtained by the chlorination of 2-exo, 10,10-trichlorobornane 1. The X ray structural analysis of 2 revealed an unusual tricyclic structure, where the two chlorine atoms occupying endo-positions are in close spatial proximity with each other and near to the neighbouring CHCl2 group. Further, it revealed that the symmetry of the molecule is distorted. The 1H and 13C NMR spectra of 2 have been assigned by means of 1H, 1H double-quantum filtered correlation spectroscopy (DQF COSY), PFG 1H, 13C HMQC (pulsed field gradient heteronuclear multiple quantum coherence), 1H, 13C heteronuclear multiple bond correlation (HMBC) experiments, and computer aided 1H NMR spectral analysis. The asymmetry of 2 is also discernible on the 1H NMR parameters. In addition, gas chromatographic (GC) properties and electron impact (EI) mass spectrum of 2 has been studied. Ab initio Hartree-Fock (HF) method with the basis set 6-31G(d) has been used for the optimization of the equilibrium geometry and calculation of total energy for 2. The optimized geometry is in good agreement with the crystal structure. According to the rotation energy profile calculated at the HF/6-31G(d) level, rotation of the chloromethyl and dichloromethyl groups are highly unlikely at the room temperature. PMID- 11482656 TI - Environmental occurrence of nitrotriphenylene observed in airborne particulate matter. AB - 1- and 2-Nitrotriphenylenes were found in the airborne particulate matter extracts collected in central Tokyo between the winter of 1998 and the winter of 1999. In particular, we have identified and quantified nitrotriphenylenes in the airborne particulate matter extracts collected over four consecutive 6-h periods on 2 December 1999. The concentrations of 1- and 2-nitrotriphenylene ranged from 0.04 to 0.44 and from 0.02 to 0.47 ng/m3, respectively, and the concentrations in the airborne particulate matter extracts collected during the 18:00-24:00 h time period were the highest of the four collection periods. 1-Nitropyrene and 2 nitrofluoranthene were also identified and quantified in the four 6-h samples. Although the concentrations of 1- and 2-nitrotriphenylenes were not higher than that of 2-nitrofluoranthene except during the 18:00-24:00 h time period, the concentrations were much higher than that of 1-nitropyrene during the four collection periods. The higher concentrations of 1- and 2-nitrotriphenylenes during the 18:00-24:00 h time period are presumably responsible for the high reactivity of parent triphenylene with NO2/NO3/N2O5, and high stability of 1- and 2-nitrotriphenylenes toward O3 + O2. In addition, the observed isomer distribution of nitrotriphenylenes suggested that direct emission of nitrotriphenylenes is also a source as well as their atmospheric formation. PMID- 11482657 TI - Bioaccumulation of cesium-137 by biota in different aquatic environments. AB - Macroalgae, isopods and fish species were exposed to 137Cs in brackish and sea water conditions for 18 days to determine radionuclide concentration factors. The concentration factors of 137Cs in brown shrimp and polychaete species were also investigated under brackish water conditions. At equilibrium, the concentration factors in macroalgae, isopod, fish, brown shrimp and polychate samples were found to be 2.5, 33, 2, 16 and 11 at 16 degrees C in brackish water conditions, respectively. The accumulation rate in macroalgae species was influenced by temperatures between 6 degrees C and 16 degrees C. The bioaccumulation of 137Cs in isopods at low salinity regime was increased significantly. At the same time, the bioaccumulation rate in macroalgae species also showed slight increase at low salinity. On the other hand, the bioaccumulation rate of 137Cs in the fish species in sea water was higher than in brackish water. PMID- 11482658 TI - Oxidation of triphenylarsine to triphenylarsineoxide by Trichoderma harzianum and other fungi. AB - Arsenic resistant strains of bacteria and fungi were isolated from soil contaminated by chemical warfare agents. Until now, no metabolic products of microbial attack against the phenyl residues of the model substrate triphenylarsine (TP) were found if it was incubated together with these strains in liquid culture assays. However, one of the isolated fungi, Trichoderma harzianum As 11, was found to oxidize TP to triphenylarsineoxide (TPO). The yeast Trichosporon mucoides SBUG 801 and the white-rot fungus Phanerochaete chrysosporium were also able to oxidize the As(III) in TP. In addition, P. chrysosporium transformed phenylarsineoxide (PAO) to phenylarsonic acid (PAA) under O2-atmosphere. By means of a respirometer system, the oxidation of TP by T. harzianum As 11 was confirmed by a significantly higher consumption of oxygen in the presence of these compounds. HPLC analysis of the oxidation products TPO and PAA in the medium of the assays provided evidence for the transfer reaction of As(III) to As(V) in organic bonds. The oxidation products TPO and PAA are more hydrophilic than TP and PAO. Therefore, it was concluded that particular fungi contribute to the mobilization of arsenic in soil contaminated by chemical warfare agents. PMID- 11482659 TI - Determination of aldehydes and ketones in air samples using cryotrapping sampling. AB - A cryotrapping sampling technique using glass traps cooled in liquid nitrogen for monitoring carbonyl compounds in air has been developed. Sampling was followed by derivatization by addition of acidified 2,4-dinitrophenyl hydrazine (DNPH) solution to the traps and an aliquot of the sample was analysed with a high performance liquid chromatograph system (HPLC), equipped with a diode array detector. The procedure was optimised concerning derivatization conditions and analytical parameters on formaldehyde, acetaldehyde, propanal, acetone, butanal and benzaldehyde. The technique was applied in monitoring their concentration in the urban atmosphere in Ljubljana. PMID- 11482660 TI - Characterization of mucilage aggregates in Adriatic and Tyrrhenian Sea: structure similarities between mucilage samples and the insoluble fractions of marine humic substance. AB - The appearance of gelatinous aggregates called mucilages causes serious damages to tourism and fishery industries of the Adriatic Sea. So, many studies have been planned and some of them are still in progress to clarify the origin and causes of the phenomenon. The scientific research has showed that mucilages are produced by several marine organisms when peculiar climatic and trophic conditions occur. Moreover, as far as the mucilage composition is concerned, although it is well known that polysaccharides give a high contribution, knowledge of the structural characteristics of mucilages and their relationship with the natural organic matter of the marine environment has not been clarified yet. In this paper a study on the characterization of the marine mucilage samples collected in the Adriatic and Tyrrhenian Seas is described. The study was performed by spectroscopic (infrared and colorimetric) techniques, and elemental analysis. The results showed that mucilage samples have chemical and structural similarities with the insoluble fraction of the marine humic substance (humin). According to experimental evidences it is possible to establish the relationship between mucilages and the dissolved organic matter (DOM) in the marine environment in order to identify the most likely pathways of mucilage formation. PMID- 11482661 TI - Degradability of selected azo dye metabolites in activated sludge systems. AB - The stability of eight environmentally relevant azo dye metabolites [o aminotoluene (2), 4,4'-thiodianiline (4), 4,4'-diaminodiphenylmethane (6), p chloroaniline (7), 2,4-toluylenediamine (9), p-kresidine (14), 2,4-diaminoanisole (15), and 2-naphthylamine (18)] was investigated in activated sludge systems and compared to their hydrolysis stability. For both studies, test systems of the EC and EPA were used. The results show that degradation under aerobic conditions proceeds via oxidation of the substituents located on the aromatic ring or on the side chain. Under anaerobic conditions, the azo bond is reductively cleaved, which leads to the substituted amines. These are toxic and potentially hazardous to the environment. PMID- 11482662 TI - Concentration of organochlorine pesticides in wine corks. AB - Wine corks were extracted and analyzed for 23 organochlorine pesticides, including alpha-, beta-, and gamma-hexachlorocyclohexanes (HCH), hexachlorobenzene, DDT, DDE and DDD, chlordane, endosulfan, dieldrin, aldrin, and endrin. This was done to investigate the occurrence, concentrations, composition profiles, and possible sources of organochlorine contamination. All groups of compounds were detected in every sample investigated, with the exception of aldrin and endrin. The total concentrations of organochlorine compounds in all samples ranged from 75-120 ng/g lipid, and for most compounds, the concentrations of organochlorines in cork were consistent with published data for other plant tissues. Differences in the relative abundances of the various classes of organochlorine pesticides were substantial and were probably due to differences in the pesticide usage practices of the various regions in which the cork producing trees were grown. PMID- 11482663 TI - Humic substances as catalysts in condensation reactions. AB - Humic substances (HS) demonstrate appreciable impact on the rate of the condensation reactions as shown in the example of the reaction between hydrazine and 4-(dimethylamino)-benzaldhyde in an aquatic environment. The catalytic activity of HS has also been demonstrated in Knoevenagel and Claisen-Schmidt reactions for condensation of carbonyl compounds with CH acids. The aquatic fulvic acids are the most active in these reactions. The velocity of the studied reactions also depends on pH, temperature, the concentration and origin of HS used. A possible micellar and acid-base catalysis mechanism in aquatic media has been suggested. PMID- 11482664 TI - Occurrence of organo-arsenicals in jellyfishes and their mucus. AB - Water-soluble arsenic compound fractions were extracted from seven species of jellyfishes and subjected to analysis by high-performance liquid chromatography inductively coupled plasma mass spectrometry (HPLC-ICP-MS) for arsenicals. A low content of arsenic was found to be the characteristic of jellyfish. Arsenobetaine (AB) was the major arsenic compound without exception in the tissues of the jellyfish species and mucus-blobs collected from some of them. Although the arsenic content in Beroe cucumis, which preys on Bolinopsis mikado, was more than 13 times that in B. mikado, the chromatograms of these two species were similar in the distribution pattern of arsenicals. The nine species of jellyfishes including two species treated in the previous paper can be classified into arsenocholine (AC)-rich and AC-poor species. Jellyfishes belonging to Semaostamae were classified as AC-rich species. PMID- 11482665 TI - TiO2-catalyzed photooxidation of arsenite to arsenate in aqueous samples. AB - The TiO2-catalyzed photooxidation of arsenite (As(III)) to arsenate (As(V)) was studied in aqueous TiO2 suspensions using a solar simulator which emitted ultraviolet and visible radiations. The concentration of As(III) was varied between 50 microg l(-1) and 10 mg l(-1), and the concentration of TiO2 between 1 mg l(-1) and 50 mg l(-1). Total oxidation of As(III) to As(V) occurred within minutes. The concentration of As(III) declined exponentially which indicates first-order kinetics. In the pH range between 5 and 9 there was no significant influence of the pH of the suspension on the reaction rate. Batch experiments without irradiation showed that part of the arsenic was adsorbed on the TiO2 surface. When using 100 microg l(-1) As and between 1 mg l(-1) and 50 mg l(-1) TiO2, 8-39% of As(III) and up to 73% of As(V) were adsorbed by TiO2. As(III) was also oxidized by UV radiation in the absence of TiO2, but the reaction was slower than in the presence of TiO2 resulting in an irradiation time too long for practical use. In addition, oxidation of As(III) in the presence of TiO2 was also observed under solar irradiation within a few minutes. PMID- 11482666 TI - Solubility of nonylphenol and nonylphenol ethoxylates. On the possible role of micelles. AB - The water solubility of nonylphenol (NP) has been estimated to be 4.9 +/- 0.4 mg/l corresponding to (2.22 +/- 0.18) x 10(-5) mol/l at 25 degrees C using shake flask and surface tension techniques. The low solubility in combination with an observed rather slow dissolution process will limit the leachability of NP in the terrestrial environment. Based on indirect evidence, it is suggested that NP, in contrast to nonylphenol ethoxylate (NPEO) with, e.g., 12 ethoxylate moieties, is not subject to micelle formation, and as such does not constitute a potential vehicle for the transport of hydrophobic pollutants in the environment. For NPEOs with a very high number of ethoxy moieties, e.g., 100, the compounds appear water soluble without micelle formation. PMID- 11482667 TI - An analytically solved kinetic model for pesticide degradation in single compartment systems. AB - An analytical kinetic model was developed to simulate the degradation of pesticides in systems such as soil or water. Based on a single compartment system, a set of simultaneous first-order differential equations was analytically solved by the eigenvalue and eigenvector method. The developed model is capable of simulating the concentrations of parent compound and any net of degradation products connected by irreversible reactions. PMID- 11482668 TI - Aerobic and anaerobic abiotic oxidation of squalene in the presence of hydroperoxides. AB - Abiotic oxidation of squalene in the presence of hydroperoxysterols was studied in seawater under aerobic and anaerobic conditions. This ubiquitous isoprenoid alkene is quickly degraded in the presence of oxygen and its oxidation results mainly in the production of tertiary alcohols and to a lesser extent of epoxides and secondary alcohols. Although the degradation of squalene logically slows down under anaerobic conditions, a significant oxidation affording similar products than in the case of aerobic degradation has been observed. These results show that hydroperoxysterols, which seem to be well preserved in Recent sediments, could contribute to the oxidation of unsaturated lipids (such as squalene) in sedimentary environments under oxic and anoxic conditions. PMID- 11482669 TI - Sorption of 1-naphthol to soil and geologic samples with varying diagenetic properties. AB - Remediation of contaminated land requires a firm understanding of the processes that occur between xenobiotics and soil colloids. It is currently accepted that the extent of xenobiotic uptake is proportional to the carbon quantity and character of the soil or geologic sample. Previous studies have developed empirical equations to predict the extent of sorption based on the aromatic carbon content. We examined these relationships with an independent set of soil and geologic samples and 1-naphthol. The 1-naphthol sorption coefficients varied significantly (P < 0.01) among sorbents and are consistent with the diagenetic properties of the organic matter in these samples. The cross-polarization magic angle spinning (CPMAS) 13C nuclear magnetic resonance (NMR) and elemental data did not concur with the sorption data for most of the soil samples. We suggest that this contradiction may be due to a third variable, the physical organization of the organic matter. Chemical methods measure the whole sample, whereas short term sorption occurs on the surface; therefore, only some organic matter domains in the soil are available for interaction with 1-naphthol. Hence, chemical data alone may be insufficient for predicting the sorption behavior of xenobiotics in soil and geologic samples. PMID- 11482670 TI - Uptake and mobility of uranium in black oaks: implications for biomonitoring depleted uranium-contaminated groundwater. AB - In a preliminary study, the uptake and the mobility of uranium (U) by black oak trees (Quercus velutina) were assessed by measuring the isotopic composition of tree rings in two mature oak trees in a heavy metal contaminated bog in Concord, MA. The bog is adjacent to a nuclear industrial facility that has been processing depleted uranium (DU) since 1959. Over the past 40 years, DU has been leaking from an onsite holding basin and cooling pond down gradient to the bog where the oaks are located. Because DU has no source outside the nuclear industry, contamination from the industrial facility is readily discernable from uptake of natural U by measuring isotopic compositions. Isotope ratio analysis confirms the occurrence of DU in bark, sapwood and heartwood tree rings dating back to 1937, pre-dating the introduction of DU at the site by at least 20 years. Isotope dilution analysis indicates high concentrations of U (>3 ppb) in sapwood that drop rapidly to relatively constant concentrations (0.3-0.4 ppb) in heartwood. These data indicate that once incorporated into tree cells, U is mobile, possibly by diffusion through the tree wood. Concentrations of U in sapwood are approximately equal to average U concentrations in groundwater onsite over the past 10 years, suggesting that oak trees can be used as present-day bioindicators of U-contaminated groundwater. We suggest that regional sampling of oak bark and sapwood is a reasonable, inexpensive alternative to drilling wells to monitor shallow groundwater U contamination. PMID- 11482671 TI - Effect of drying and composting biosolids on the movement of nitrate and phosphate through repacked soil columns under steady-state hydrological conditions. AB - Effects of two "enhanced" treatments (drying and composting mesophilic anaerobically digested (MAD) biosolid) on nutrient leaching were investigated. Repacked sandy or sandy loam textured soil cores amended with fresh, dried and composted MAD biosolid (250 kg N ha(-1)), were investigated under steady-state hydrological conditions. Two 24 h, 4.5 mm h(-1) rainfall events, with a 14-day interval, were simulated using water-tracers. Losses of nitrate from the sandy loam soil during rainfall event 1 (43.9-68.0 mg kg(-1)) were significantly greater (P < or = 0.05) than during event 2 (6.4-11.9 mg kg(-1)). Phosphate losses were significantly greater (P < or = 0.05) during event 2 (up to 0.30 mg kg(-1)) compared to the first (< 0.05 mg kg(-1)). The sand soil showed similar effects. Losses of nitrate-N (percentage of total N applied) from the sand soil were small (around 0.06% for fresh/dried and 0.63% for composted MAD biosolids). Losses of nitrate-N from the sandy loam soil were greater; 4% for fresh and dried and 3% for composted MAD biosolids. This research showed that drying MAD biosolid had little impact on nitrate and phosphate losses from soil compared to fresh MAD biosolid. The effect of composting MAD biosolid on nutrient losses was more variable. PMID- 11482672 TI - Enhanced transport of pesticides in a field trial with treated sewage sludge. AB - This study was designed to provide high-density data on spatial distribution of three herbicides with different physiochemical characteristics in a sludge amended and non-amended control field over the course of an irrigation season. The field experiment was carried out on a sandy loam Hamra Red Mediterranean soil (Rhodoxeralf) at Bet Dagan, Israel. After a single 50 mm irrigation event, the mean centers of mass (COM) in the control field were at 15.6, 14.9, and 17 cm for bromacil, atrazine and terbuthylazine, respectively; in the sludge-amended field, mean COMs were at 28.8, 31.2, and 34.1 cm, respectively. After 500 mm of irrigation in the control field, the COM depth distribution of the three pesticides was inversely correlated with octanol-water (Kow) distribution coefficients and soil sorption coefficients (Koc), and positively correlated with aqueous solubilities. After 500 mm irrigation in the sludge-amended field, the mean terbuthylazine COM was at 19.8 cm versus 13.8 cm for the control field, demonstrating a sustained enhanced effect on terbuthylazine transport. Downward transport of atrazine was also enhanced by sludge amendment, albeit less than terbuthylazine. Bromacil was preferentially accumulated in the upper soil layers of the sludge-amended field as compared with the control field after 500 mm irrigation. The enhanced transport of all three pesticides in the sludge-amended field after a single irrigation event is attributed to development of preferential flow pathways around hydrophobic clods of sludge. Enhanced transport of terbuthylazine, and to a lesser extent, atrazine, throughout the irrigation season, is attributed to their transport as complexes with dissolved, colloidal and suspended organic matter derived from sludge degradation. PMID- 11482673 TI - Instrumental recording and biomonitoring of ambient ozone in the Greek countryside. AB - Among eight commercial Greek varieties of tobacco (Nicotiana tabacum L.) tested for their ozone-sensitivity levels, the Zichnomirodata (KK6/5) variety was found to be the most sensitive, although less sensitive than the well-known super sensitive Bel-W3. Besides qualitative differences in the appearance of macroscopic symptoms these two varieties can be used simultaneously as a reliable pair of ozone bioindicators. The occurrence of ozone in the Greek countryside was surveyed by biomonitoring in 14 rural regions over the country and by a simultaneous biomonitoring and instrumental recording of ozone concentrations at a single remote side (Pournaria, Arcadia). Phytotoxic symptoms were observed mainly on the leaves of Bel-W3 and occasionally on those of Zichnomirodata varieties, suggesting that ozone levels were high enough to affect at least sensitive species. The instrumental monitoring (during a total period of 912 h) revealed maximum hourly O3 concentration 62 ppb, while the thresholds of 30, 40 and 50 ppb were exceeded for 40%, 20% and 6% of the recording period, respectively. The accumulated exposure over 40 ppb (AOT40) for the daylight hours over the 38 monitored days was 680 ppb h. PMID- 11482674 TI - Biodegradability of ethylenediamine-based complexing agents and related compounds. AB - The biological degradability (Zahn-Wellens test) of ethylenediamine derivatives with carboxymethyl and 2-hydroxyethyl groups was investigated. Mixed bacterial culture (activated sludge) was used as inoculum (non-adapted sludge and sludge adapted at different mean biomass retention time, the so-called sludge age). Biodegradability of ethylene(propylene)di(tri)amine-based complexing agents depends on the character and number of substituents and nitrogen atoms in the molecule. Tetra(penta)substituted derivatives with two or more tertiary nitrogen atoms and carboxymethyl or 2-hydroxyethyl groups in the molecule (EDTA, DTPA, PDTA, HEDTA) are very stable from an environmental point of view. On the contrary, disubstituted derivatives with two secondary nitrogen atoms in the molecule (e.g., EDDA) are potentially degradable. PMID- 11482675 TI - Photochemically induced formation of the "Al13" tridecameric polycation in the presence of Fe(III) and organic acids. AB - We found that light-induced Fe(III) reduction associated with the oxidation of a simple hydroxy-carboxylic acid (lactate) caused the formation of the AlO4Al12(OH)24(H2O)12 polycation ("Al13"). Initial conditions were a lactate:Al:Fe ratio of 1:0.76:0.11 in a partially neutralized solution. Base was added rapidly and no Al13 was detected in samples kept in the dark. With exposure to light, Fe(III) reduction was rapid and Fe(II) reached a maximum within 1 day. After the maximum, steady-state Fe(II) declined from 54% to 43% over eight days. During this same time period, the lactate concentration fell to 2% of the original, pH rose from 4.05 to 4.46, and the Al13 detectable by 27Al NMR increased to 2.3 mmol l(-1) (51% of the total solution Al). The formation of Al13 is attributed to the pH rise resulting from the removal of the organic acid buffer. Similar photo-induced chemical changes occur in natural waters and may promote the formation of Al13, conditions permitting. PMID- 11482676 TI - Trace metals in different species of mollusca, water and sediments from Taiwan coastal area. AB - Since October 1994, a long-term program of Asia/Pacific Mussel Watch: Taiwan Regional Studies has been carried out. The results indicate that trace metal contents in mollusca varied among 30 different species and the environments (water and sediments) along the Taiwan coast. The orders of bioaccumulation of trace metals in mollusca were: Cu (over 200 microg/g), Thais clavigera > Isognomon legumen > Clibanarius rivescens > Crassostrea gigas; Zn (over 700 microg/g), Moruta granulata > C. gigas > Asiatica cypraea arabica > T. clavigera; Cd (over 5 microg/g), Trochus hanleyanus > Acanthopleura japonica > Nerita albicilla > Patella flexuosa; Pb (over 2 microg/g), P. flexuosa > C. gigas > T. hanleyanus > T. clavigera > C. gigas > Capiyulum mitella; Ni (over 10 microg/g), Meretrix lusoria > Philine sp. > Littoraria scabra > Tridacna squamosa > T. hanleyanus; Cr (over 30 microg/g), Littoraria undulata > T. hanleyanus > N. albicilla > Nerita chamaelor > M. granulata; As (over 20 microg/g), Perna viridis > L. scabra; and Sn (over 5 microg/g), P. viridis > L. undulata> C. mitella> C. gigas. Their seasonal and regional variations as well as their correlation are evaluated and discussed. PMID- 11482677 TI - Fugacity superposition: a new approach to dynamic multimedia fate modeling. AB - The fugacities, concentrations, or inventories of pollutants in environmental compartments as determined by multimedia environmental fate models of the Mackay type can be superimposed on each other. This is true for both steady-state (level III) and dynamic (level IV) models. Any problem in multimedia fate models with linear, time-invariant transfer and transformation coefficients can be solved through a superposition of a set of n independent solutions to a set of coupled, homogeneous first-order differential equations, where n is the number of compartments in the model. For initial condition problems in dynamic models, the initial inventories can be separated, e.g. by a compartment. The solution is obtained by adding the single-compartment solutions. For time-varying emissions, a convolution integral is used to superimpose solutions. The advantage of this approach is that the differential equations have to be solved only once. No numeric integration is required. Alternatively, the dynamic model can be simplified to algebraic equations using the Laplace transform. For time-varying emissions, the Laplace transform of the model equations is simply multiplied with the Laplace transform of the emission profile. It is also shown that the time integrated inventories of the initial conditions problems are the same as the inventories in the steady-state problem. This implies that important properties of pollutants such as potential dose, persistence, and characteristic travel distance can be derived from the steady state. PMID- 11482678 TI - The solubility of gases and vapours in dry octan-1-ol at 298 K. AB - Ostwald solubility coefficients of 74 compounds in dry octan-1-ol at 298 K have been determined, and have been combined with literature values and additional values we have calculated from solubilities in dry octan-1-ol and vapour pressures to yield a total of 161 log L(OctOH) values at 298 K. These L(OctOH) values are identical to gas-to-dry octan-1-ol partition coefficients, often denoted as K(OA). Application of the solvation equation of Abraham to 124 values as a training set yielded a correlation equation with n = 124, S.D. = 0.125, r2 = 0.9970 and F = 7731. This equation was then used to predict 32 values of log L(OctOH) as a test set, giving a standard deviation, S.D. of 0.131, an average absolute deviation of 0.085 and an average deviation of -0.009 log units. The solvation equation for the combined 156 log L(OctOH) values was log L(OctOH) = 0.120 - 0.203R2 + 0.560pi2(H) + 3.560 sum(alpha2(H)) + 0.702 sum(beta2(H)) + 0.939 logL16, n =156, r2 = 0.9972, S.D. = 0.125, F = 10573, where, n is the number of data points (solutes), r the correlation coefficient, S.D. the standard deviation and F is the F-statistic. The independent variables are solute descriptors as follows: R2 is an excess molar refraction, pi2(H) the dipolarity/polarisability, sum(alpha2(H)) the overall or summation hydrogen-bond acidity, sum(beta2(H)) the overall or summation hydrogen-bond basicity and L16 is the Ostwald solubility coefficient on hexadecane at 298 K. The equation is consistent with similar equations for the solubility of gases and vapours into methanol, ethanol and propan-1-ol. It is suggested that the equation can be used to predict further values of log L(OctOH), for which the solute descriptors are known, to within 0.13 log units. PMID- 11482679 TI - Primary biodegradation of veterinary antibiotics in aerobic and anaerobic surface water simulation systems. AB - The primary aerobic and anaerobic biodegradability at intermediate concentrations (50-5000 microg/l) of the antibiotics olaquindox (OLA), metronidazole (MET), tylosin (TYL) and oxytetracycline (OTC) was studied in a simple shake flask system simulating the conditions in surface waters. The purpose of the study was to provide rate data for primary biodegradation in the scenario where antibiotics pollute surface waters as a result of run-off from arable land. The source of antibiotics may be application of manure as fertilizer or excreta of grazing animals. Assuming first-order degradation kinetics, ranges of half-lives for aerobic degradation of the four antibiotics studied were 4-8 days (OLA), 9.5-40 days (TYL), 14-104 days (MET) and 42-46 days (OTC). OLA and OTC were degraded with no initial lag phase whereas lag phases from 2 to 34 days (MET) and 31 to 40 days (TYL) were observed for other substances. The biodegradation behaviour was influenced by neither the concentrations of antibiotics nor the time of the year and location for sampling of surface water. Addition of 1 g/l of sediment or 3 mg/l of activated sludge from wastewater treatment increased the biodegradation potential which is believed to be the result of increased bacterial concentration in the test solution. Biodegradation was significantly slower in tests conducted in absence of oxygen. Assessments of the toxic properties of antibiotics by studying the influence on the biodegradation rates of 14C-aniline at different concentrations of antibiotics showed that no tests were conducted at toxic concentrations. PMID- 11482680 TI - Dioxin and dioxin-like PCB impurities in some Japanese agrochemical formulations. AB - The profile and amount of dioxin impurity in agrochemicals were studied through detailed analysis of historic Japanese formulations. The chemicals analyzed include pentachlorophenol (PCP), 2,4,6-trichlorophenyl-4'-nitrophenyl ether (chloronitrofen, CNP), 2,4-dichlorophenyl-4'-nitrophenyl ether (nitrofen, NIP), tetrachloro-iso-phthalonitrile (chlorothalonil, TPN), 2-methyl-4-chloro phenoxyacetic acid (MCP) and 2,4-dichlorophenoxyacetic acid (2,4-D). Among the six, two herbicides, PCP and CNP, produced during the 1960s and 1970s, contained very high concentrations of PCDD/DFs and TEQ. Others contained relatively low concentrations of PCDD/DFs. Dioxin-like PCB concentrations in all chemicals studied were low and their contributions to TEQ were negligible. The total dioxin emissions from the use of agrochemicals in Japan during the past 40 years (1955 1995) were estimated to be about a few hundred thousand kg of PCDD/DFs and 250 kg of WHO-TEQ from PCP and 190 x 10(3) kg of PCDD/DFs and 440 kg of WHO-TEQ from CNP. The major dioxin congeners present in PCP formulations were highly chlorinated PCDD/DFs that can be formed by the coupling of PCP and/or 2,3,4,6 tetrachlorophenol, and those in the CNP formulations were tetra- to hexa chlorinated PCDD/DFs that can be formed from 2,4,6-trichlorophenol and/or 2,3,4,6 tetrachlorophenol. PMID- 11482681 TI - Effect of sludge-amendment or nutrient addition on the biodegradation of the herbicide isoproturon in soil. AB - Adding sludge to agricultural soil results in added organic matter, nutrients and metallic and/or organic pollutants. These components may modify the behaviour of pesticides in the soil. We monitored possible changes in the degradation of the herbicide isoproturon (production of CO2 and degradation products) in soil amended with sludge, heavy metals or nitrogen and phosphorus. The treated and control soils were incubated under controlled conditions for 60 days. The nitrogen and phosphorus had the greatest effect on isoproturon degradation, independent of the presence of pollutants. Mineralisation of the herbicide to CO2 was slow and seemed to be linked to a fast degradation and to the accumulation of a complex degradation product that was neither catabolized nor adsorbed, 4,4' diisopropylazobenzene. This degradation pathway also produced smaller amounts of non-extractable residues. Sewage sludge had no significant effect on isoproturon degradation, despite a large increase of organic matter mineralisation (factor 2). PMID- 11482682 TI - Degradation of 14C-carbofuran in soil using a continuous flow system. AB - 14C-carbofuran underwent considerable mineralization (approximately 30% of the applied activity) in Vertisol soil under moist and flooded conditions during 60 days incubation. Bound residues were formed under both the conditions, the extent being more in moist soils (approximately 55% of the applied activity) than under flooded conditions (approximately 41% of the applied activity). 3-Keto carbofuran was the only significant metabolite observed under flooded conditions. PMID- 11482683 TI - Boiling points of halogenated organic compounds. AB - The normal boiling points of a number of halogenated organic compounds have been compiled from experimental measurements over three decades. Some of these chemicals have not been reported in the literature. The substances listed are halogenated aliphatic hydrocarbons, halogenated aliphatic ethers, halogenated ring (cyclic) hydrocarbons and other related compounds. PMID- 11482684 TI - Burn wound itch control using H1 and H2 antagonists. AB - This study investigated the use of a combination of H1 and H2 antagonists and topical medications to control burn wound itch. Graeco-Latin square assignment provided an oral combination of 1) cetirizine and cimetidine or 2) diphenhydramine and placebo in four divided doses. The study protocol lasted 16 days divided into 4-day intervals, scoring itch before the initial dose of medication and at 1-hour, 6-hour, and 12-hour intervals after the first medication. A significant difference between mean itch scores across the four times was observed (Wilks' Lambda F = 26.52, df = 3, P <.0005). A three-way nested repeated measures interaction effect (Wilks' Lambda F = 9.85, df = 9, P <.0005) was observed representing a significantly different pattern on days 1 to 4 of the study compared with the remaining days. Controlling for the effect of topical medications, the cetirizine/cimetidine combination demonstrated a dramatic improvement at 1 and 6 hours, and a moderate improvement at 12 hours after initial medication for the day when compared with the diphenhydramine/placebo combination. PMID- 11482685 TI - Socioeconomic and cultural influence in the causation of burns in the urban children of Bangladesh. AB - This prospective case-control study was conducted on 105 consecutive cases admitted to the burn units of Dhaka Shishu Hospital and Dhaka Medical College Hospital. The same number of controls were selected randomly from the community and matched with respect to age group, sex, and location of residence within the defined metropolitan area during the period January to September 1997. Both the controls and cases were divided into three age groups. Group A (19 controls and 19 cases) included neonates and infants, group B included children from 1 to 4 years, and group C included children from 4 to 12 years. The mean age of the controls in group A was 8.42+/-3.86 months and in group B and C was 5.72+/-4.05 years. The mean age of cases in group A was 4.79+/-4.05 months and in group B and C was 5.08+/-2.87 years. A scoring system was used to analyze 14 socioeconomic and cultural factors, which indicated highly significant differences between controls and cases (P < 0.001), and also between male (P < 0.001) and female (P < 0.001) controls and cases. There were significant differences between controls and cases in group A (P < 0.001), group B (P < 0.01), and group C (P < 0.001). Differences were not significant between males and females within groups A, B, and C for cases or controls (P > 0.05). Regression analysis of the total scores for cases and controls indicated a negative correlation between burns and the socioeconomic variables included in the study. There were highly significant associations between burns and lack of alertness to burns among parents (P < 0.001), clothing of manmade fabrics (P < 0.001), and cooking equipment in the kitchen within reach of children (P < 0.001). There was a significant association between burns and illiteracy of the mother (P < 0.01), housing located in slums and congested areas (P < 0.01), illiteracy of the father (P < 0.02), presence of preexisting impairment in children (P < 0.05), presence of a history of burns among siblings (P < 0.05), and low economic status of the parents (P < 0.05). No significant associations were indicated between controls and cases with respect to number of children in the family (P > 0.1), family pattern (P > 0.1), conjugal bonding between parents (P > 0.5), and sibling's death and disability from burns (P > 0.5). It is evident that an increase in alertness to burns, easy availability of clothing of natural fabrics, keeping cooking equipment beyond the reach of the children, increased literacy among parents and children, and improvement in family income might decrease the incidence of burns in children. PMID- 11482686 TI - The use of TENS for pruritus relief in the burns patient: an individual case report. AB - The case report outlines the use of transcutaneous electrical nerve stimulation (TENS) for the relief of pruritus in a 19-year-old patient who had sustained 70% mixed-thickness flame burns. Once healed, two electrodes were applied to the region of most severe itching, which was the lumbar spine. They were applied at 10 AM and remained in place until he went to bed. A visual analog scale (VAS) rating of itching was used to measure the sensation subjectively before application, immediately after application, and 4 hours after continuous application. Readings were taken for 5 consecutive days and then followed up during a 2-week period. At this time the patient discontinued the use of TENS, because itching had decreased significantly. Results showed that between day 1 and day 16 there was a 50% improvement before application and a 100% improvement 4 hours after application of TENS. These results suggest that a study to investigate the effect of TENS for the relief of pruritus in the burn patient would be justified to provide an alternative, noninvasive treatment approach. PMID- 11482687 TI - In search of effective education in burn and fire prevention. AB - Children younger than 14 years continue to be a high-risk group for burn-related and fire-related injuries. Burn and fire educators must find a way to reach these children that captures their imaginations. There may be no better way than games. Two burn and fire prevention games were developed. The games were distributed to 38 school districts encompassing a total of 164 elementary schools and reaching more than 1,035 youngsters in grades 1 through 4 in a two-county community. Before playing each game, the participants completed a multichoice pretest. A similar posttest was administered after gaming to determine mastery and retention of knowledge. In addition, classroom instructors were given an evaluation form to assess content, quality, and effectiveness. Pretest and posttest results indicated students gained and retained significant knowledge. Instructor evaluation recognized these games as entertaining and exciting, precipitating additional questions leading to further classroom discussion and learning. PMID- 11482688 TI - Does the addition of nystatin to 5% mafenide acetate and genitourinary irrigant solutions interfere with their antimicrobial activity? Assessment by two topical antimicrobial test assay systems. AB - Results previously reported using the Wet Disc Topical Antimicrobial Assay (WDA) suggested that adding nystatin (NY) to a 0.5% mafenide acetate (MA) suspension or genitourinary irrigant (double antibiotic [DAB]) to expand their antimicrobial activity to include Candida sp. antagonized the antibacterial effect of MA but not DAB. We use DAB solution as described by the authors of the previous study, also, but we use a 5% commercially available mafenide acetate solution instead of the in-house prepared 0.5% mafenide acetate suspension that they used. Further, we use both the WDA and the Agar Well Diffusion Topical Antimicrobial Assay (AWDA) to test topical antimicrobials at this institution. In light of the previously reported results, this study 1) examined whether adding nystatin to DAB or the 5% mafenide acetate solution used at this institution caused any interference in the ability of these substances to migrate through the agar matrices and cause zones of growth inhibition in the two test assay systems and 2) compared the assessment of microbial susceptibility (by very precise definition) between the two systems. The addition of nystatin did not interfere with the ability of either DAB or mafenide acetate to migrate through the agar matrices and cause clear zones. However, on the assessment of susceptibility a significantly larger number of organisms were judged susceptible using the AWDA than the WDA. We believe that the disparity is caused by a large difference in agar diffusion kinetics between the two assays. Therefore, we recommend the AWDA rather than the WDA for susceptibility studies. PMID- 11482689 TI - Fate of free muscle transfer covering chronically infected burned skull. AB - This case report describes a 28-year-old male who sustained a 46,000-V cranial electrical injury that resulted in devascularization, necrosis, and secondary infection of the skull despite bone coverage with a free muscle transfer. In the face of osteomyelitis of the skull, the free muscle transfer and the brain were not compromised. A review of the literature and this case reinforce the practice of leaving the skull in place and covering it with vascularized tissue. Resection of the infected skull and later replacement with alloplastic material were curative. PMID- 11482690 TI - Characteristics of burn patients developing pressure ulcers. AB - In 1997 a prospective study to identify pressure ulcer development and identify risk factors was conducted on 217 consecutive patients in the burn center. Weekly rounds were conducted, and the specifics of pressure ulcer development, demographics, concurrent injuries, comorbid conditions, risk factors, and patient outcomes were identified. An incidence of 4.1% was identified, with the most common locations being the sacrum, occipital area, and heel. All patients with pressure ulcers met criteria for a major burn injury. The patients had concurrent injuries or an infectious process, had more surgical procedures, and were older than the group that did not develop pressure ulcers. A lower Braden Scale for risk assessment and a later onset of ulceration was noted in this population compared with previous studies of nonburn patients. The results of this study have led to changes in our burn clinical pathways that target preventive strategies in high-risk patients. PMID- 11482691 TI - Adolescents' perceptions of participation in a burn patient support group. AB - This study was designed to access the discerned value of the group experience for participants in a voluntary, open group led by a mental health professional to acute burn and reconstructive adolescent surgical inpatients and outpatients. It assessed the perceptions of participants about the reasons for their group attendance, therapeutic gains achieved from this experience, and their feelings about the group's value to other burn survivors. The responses validated the importance of using group support for burn survivors. Gains cited were the opportunity to define for others and selves how a person copes with being burned (65%), meeting others who had similar experiences (60%), expression of feelings about having been burned (50%), and learning new coping skills (45%). Ninety percent of the respondents would recommend the group to others and stated they would attend a similar group again. PMID- 11482692 TI - Use of nitric oxide as an adjuvant therapy in respiratory failure after burn injuries. PMID- 11482693 TI - Genesis of cerebral aneurysms--an update. AB - To elucidate the molecular pathogenesis of diseases has become a crucial step in the development of new treatment strategies. Although the pathogenesis of cerebral aneurysms has been studied intensively, it is poorly understood. Endogenous factors like elevated arterial blood pressure, the special anatomy of the Circle of Willis or the effect of haemodynamic factors, particularly originating at vessel bifurcation, are all known to be involved in the growth and rupture of an aneurysm. There is an ongoing discussion as to whether these factors also contribute to the very early steps of pathogenesis. Arteriosclerosis and secondary inflammatory reactions are thought to be elementary preconditions. Exogenous factors like cigarette smoking, heavy alcohol consumption or certain medications known to help generate arteriosclerosis and elevated blood pressure have also been found to be related to the occurrence of cerebral aneurysms. Furthermore, there has been a long-lasting debate on whether aneurysms might develop as a result of an inborn genetic defect. First-degree relatives of patients with cerebral aneurysms have a higher risk of having an aneurysm. In addition, the elevated prevalence of cerebral aneurysms in patients suffering from various inherited diseases points to a genetic background in the development of an aneurysm. Recent advances in molecular biology provide evidence that genetic variants of different candidate genes are associated with the occurrence of cerebral aneurysms. The aim of this review is to expose the current status of these various hypotheses and their contribution to the pathogenesis of cerebral aneurysms in order to provide a basis for future investigations in this field. PMID- 11482694 TI - Endovascular occlusion of intracranial aneurysms with Guglielmi detachable coils: correlation between coil packing density and coil compaction. AB - This retrospective analysis was undertaken to evaluate a possible relationship between coil packing density and coil compaction on intracranial aneurysms embolized using Guglielmi detachable coils (GDCs). Of the patients who underwent endovascular surgery using GDC in our hospital between 1994 and 1998, 33 patients had endovascular treatment with GDC and were examined by follow-up angiography at least 12 months after surgery. They had coil embolization to the extent where aneurysms were no longer filled or only faintly filled as shown by cerebral angiography immediately after surgery. At follow-up angiography, coil compaction was observed in 3 aneurysms. In all patients with coil compaction, the coil packing density was below 20% (14.5 +/- 4.0%). On the other hand, it was over 20% (25.7 +/- 4.7%) in all patients without coil compaction. In the 11 patients with a basilar bifurcation aneurysm, the coil packing density was over 24% and no coil compaction was observed. The coil packing density seems to be one of the critical factors, particularly for predicting whether or not coil compaction will occur. Endovascular surgery should be performed to obtain coil packing density higher than 20%. PMID- 11482695 TI - Carotid endarterectomy: a new technique replacing internal shunts. AB - BACKGROUND: Carotid endarterectomy has been reported to increase the time free from cerebral ischemic events in both symptomatic and asymptomatic patients with a high grade of stenosis of the internal carotid artery. In cases in whom the compensatory circulation during the carotid clamp time is not sufficient, the use of intraluminal shunts has been proposed. However, the use of intraluminal shunts present several problems, such as the technical difficulties in positioning the shunt, the variability of time requested for the placement, the inconstancy of the blood flow during surgery, and the need to clamp off the carotid to introduce and remove the shunt. For these reasons, most operators prefer not to employ intraluminal shunts, while others do use them only in selected cases. The purpose of this work is to present, for the first time, a new type of temporary extraluminal shunt, connecting the femoral to the internal carotid artery with the interposition of a roller pump to regulate the blood flow. This method allows one to perform carotid endarterectomy without interrupting the blood flow to the brain. METHODS: 407 consecutive patients, who underwent carotid endarterectomy between August 1992 and April 2000, were considered. 35 patients presented an absolutely insufficient collateral circulation, demonstrated by important modifications of the electroencephalographic monitoring during the carotid clamp time. In these patients the endarterectomy was performed using a new femoral carotid extraluminal shunt. FINDINGS: In all the cases in whom the femoral carotid extraluminal shunt was positioned, the normalisation of electroencephalographic monitoring was achieved by regulating the blood flow with the interposed roller pump. The use and the placement of the shunt was simple and easy. None of the patients showed postoperative complications, except for one who had a stroke two days after surgery. INTERPRETATION: The results obtained, although to be confirmed by further studies, seem to demonstrate the effectiveness of our femoral-carotid extraluminal shunt, which was simple to use and safe. PMID- 11482696 TI - Clinical features and medical treatment of male prolactinomas. AB - BACKGROUND: Prolactinomas found in male patients show distinct clinical features compared to those in female patients, which may warrant a different treatment strategy. METHOD: To clarify their clinical features and to evaluate the treatment results, specifically the results of surgical treatment and non surgical treatment solely with oral bromocriptine, we retrospectively reviewed our experience in male prolactinoma cases. FINDINGS: From 1988 to 1998, we had 184 pituitary adenoma patients, and thirteen of those were male patients with a pure prolactinoma. Of the thirteen patients, eight underwent transsphenoidal surgery followed by oral bromocriptine (surgical group), and five were treated solely with bromocriptine or terguride (non-surgical group). In both groups, the visual symptoms and signs resolved after the treatment, and the serum prolactin levels were normalised with minimal maintenance dose of bromocriptine. Notably, improvement of the visual symptom in the three non-surgically treated patients was observed within a week following the bromocriptine administration. INTERPRETATION: Although surgery would continue to play an important part of treatment in some cases with a large tumour, our experience suggests that drug treatment without surgery can be a safe and effective option in the management of male prolactinoma patients. PMID- 11482697 TI - Meningiomas of the sellar region presenting with visual impairment: impact of various prognostic factors on surgical outcome in 62 patients. AB - BACKGROUND: Meningiomas of the supra- and parasellar region can cause insidious visual loss by optic nerve compression. 62 cases with such tumours affecting the anterior optic pathways were analysed to assess the surgical results and prognostic factors with particular attention to visual outcome. METHOD: In all patients, visual deterioration was the first clinical manifestation. Eleven lesions had their origin at the anterior clinoid process, 24 at the tuberculum sellae, 10 at the planum sphenoidale, two in the optic canal, 10 in the medial sphenoidal wing, and five in the olfactory groove. All patients underwent microsurgical tumour resection. Median age at the time of operation was 54 years, median duration of symptoms seven months. The mean follow-up time was 5.2 years (range 2 to 8 years). Statistical analysis of prognostic factors (gender, age, tumour location, tumour size, duration of symptoms, brain tumour interface, resection grade, preoperative visual loss, Glascow Outcome Score) was performed using univariate and multivariate analysis. FINDINGS: The severe morbidity rate was 6.4%. Two patients died within the first 30 postoperative days. Overall, vision improved in 39 (65%) patients, in 11 (18%) it was unchanged, and worse in 10 (17%). Visual prognosis was favourably affected by age under 54 years (p < 0.025), duration of symptoms of less than seven months (p < 0.037), and the presence of an intact arachnoid membrane around the lesion (p < 0.001). Severe preoperative loss of visual acuity (<0.02) appeared to be an unfavourable prognostic factor (p < 0.047). INTERPRETATION: Possible difficulties and surgical outcome in such patients can be predicted successfully. These facts in connection with new therapeutic modalities (radiosurgery, adjuvant therapies) will demand a careful risk assessment and should influence the treatment strategies and the degree of operative aggressiveness in the future. PMID- 11482698 TI - Assessment of cure and recurrence after pituitary surgery for Cushing's disease. AB - BACKGROUND: The treatment of choice in Cushing's disease is transsphenoidal adenomectomy with a recurrence rate ranging 9-23%. We investigated whether abnormal hormonal responses may predict the relapse in "operated" patients followed-up for a long period. METHOD: Sixty-eight surgically treated patients with Cushing's disease were followed-up for 12-252 months. Forty-eight patients underwent selective adenomectomy, 17 enlarged adenomectomy and 3 underwent total hypophysectomy. After surgery ACTH and cortisol levels were measured after stimulatory (desmopressin and CRH) and inhibitory tests (dexamethasone and loperamide). FINDINGS: After operation 46 patients were cured (group A), 15 patients only normalized cortisol levels (group B), 7 patients were surgical failures. During the follow-up, a disease-free condition was maintained in 48 of 61 cases (79%), while a recurrence occurred in 13 patients (21%, 5 of group A and 8 of group B). In 5/13 patients who relapsed an absent inhibition after dexamethasone and an exaggerated response to CRH test preceded the recurrence. In 5 other patients the relapse was suspected by loperamide test. In the 3 remaining cases, positive responses to desmopressin preceded the recurrence. In 7/13 patients who relapsed the pituitary tumour was visualized by MRI/CT imaging. INTERPRETATION: During the follow-up a careful assessment of ACTH dynamics is needed. Although no single test can reliably predict the late outcome, individual patients at risk for relapse may be identified by abnormal responses to desmopressin, CRH and loperamide tests; particularly, the persistent responsiveness to desmopressin may be a criterion of risk for recurrence in patients who only normalized cortisol levels after surgery. PMID- 11482699 TI - Aggressive decompressive surgery in patients with massive hemispheric embolic cerebral infarction associated with severe brain swelling. AB - Massive hemispheric cerebral infarction, also known as malignant infarction, is characterized by rapid clinical deterioration due to brain swelling and downward transtentorial herniation, and is associated with a mortality of 80%. Early patient selection and establishment of the optimum therapeutic modality are important to improve the outcome. Early clinical, computed tomography (CT), and angiographic characteristics were analysed to identify patients with malignant infarction and external and internal decompression was performed, with unco parahippocampectomy if needed, and the outcomes were compared with those of conservative treatment. Thirty-four of 55 patients admitted with large cerebral infarctions due to embolism showed rapid clinical deterioration due to brain swelling and herniation. These 34 patients were treated under a diagnosis of malignant infarction by decompressive surgical treatment (19 cases) or conservative treatment (15 cases). CT showed significantly higher infarction volume in patients with malignant infarction (288 +/- 62 cm3) compared to patients with non-malignant infarction (200 + 57 cm3, P < 0.001) and angiography showed a higher incidence of recanalization of the occluded vessels in patients with malignant infarction (58%) compared to patients with non-malignant infarction (15%, P < 0.05). Discriminant analysis revealed that an infarction volume of more than 240 cm3 was predictive of malignant infarction with 76.4% accuracy. Basic clinical characteristics on admission and deterioration were not statistically different between the surgically treated and conservatively treated groups of patients with malignant infarction. The shift of midline structures was significantly improved (14 +/- 3.5 to 10 +/- 4.7 mm) after surgical treatment (P < 0.05). compared to deterioration (12 +/- 5.8 to 15 +/- 4.5 mm) after conservative treatment. The mortality was 67% in the conservative group and 16% in the surgical group. Surgical treatment significantly improved the mortality and Glasgow Outcome Scale score (P < 0.01). However, the mean Barthel Index scores of the survivors were not significantly different. An infarct volume of more than 240 cm3 on CT and angiographic recanalization of the occluded artery are predictors of fatal brain swelling after massive cerebral infarction. Decompressive surgical treatment dramatically improves the mortality of massive hemispheric infarction. PMID- 11482700 TI - Intracranial cerebrospinal fluid distribution and its postoperative changes in normal pressure hydrocephalus. AB - BACKGROUND: This study was conducted to investigate the usefulness of intracranial cerebrospinal fluid (CSF) volume measurement using MR-based methods in the management of patients with normal pressure hydrocephalus (NPH). METHODS: The study group comprised 19 patients with NPH who showed a favorable outcome after ventricular shunting, 15 normal volunteers (NV), and 15 patients with cerebrovascular disease (CVD). A 3D-fast asymmetric spin echo MR imaging sequence and the region-growing method were used to extract the CSF space from MR images. Ventricular volume (VV) and intracranial CSF volume (ICV) were measured and the VV/ICV ratio was calculated in each case. In NPH patients, the CSF volume was measured again after shunting. FINDINGS: The mean VV and VV/ICV ratio in the NPH group (91.1 mL and 45.2%, respectively) were significantly (p < 0.01) higher than those in the NV group (26.5 mL and 13.7%) and in the CVD group (44.5 mL and 17.8%). On the other hand, mean ICV values were not significantly different among the three groups. The VV was markedly decreased postoperatively (mean -40.7%), whereas the ICV was unchanged, resulting in a marked reduction in the VV/ICV ratio (mean -39.3%). INTERPRETATION: These results suggest that patients with NPH have a unique intracranial CSF distribution, with an enlarged VV and a slightly increased ICV, resulting in a high VV/ICV ratio. Shunting led to dramatic improvement in our patients. It is likely that CSF measurement can provide valuable information in the management of patients with NPH. PMID- 11482701 TI - Endoscopic transnasal approach to the pituitary lesions using navigation system (InstaTrak System). AB - Endoscopic transnasal approach has become a procedure of choice for the surgical management of pituitary lesions. However, in conventional endoscopic transnasal surgery, the surgeon may become disorientated to the actual operating position. In our series, 47 patients have undergone an endoscopic transnasal approach to the pituitary with the use of the navigation system called InstaTrak for real time imaging. This image guidance system proved valuable for anatomical localization during pituitary surgery. We have reduced the average surgical time, and improved patient outcome. As a consequence, complications during surgery should decrease and safety should increase. Intra-operative image guidance is expected to have major advantageous effects on pituitary surgery by allowing the surgeon to remove lesions more efficiently. As this system is improved technically and surgeons become more proficient in their use, there should be better postoperative outcomes. PMID- 11482702 TI - The Department of Neurosurgery at the University Hospital of Essen Medical School. PMID- 11482703 TI - Combined surgery and endovascular stenting for basilar artery stenosis refractory to balloon angioplasty: technical case report. AB - The authors report a case of symptomatic basilar artery stenosis treated by stenting via the surgically exposed C1 vertebral artery. This case was initially treated by percutaneous transluminal angioplasty via a transfemoral route but resulted in unsatisfactory dilatation. Stenting via a transfemoral route also resulted in failure because of the coiling of the proximal vertebral artery. Direct puncture of the vertebral artery beyond the coiling portion was tried but a stent could not be delivered beyond the C2 vertebrae. Finally, the vertebral artery was surgically exposed between C1 and the occipital bone and a stent was introduced into the lesion from this portion under fluoroscopic control. The basilar artery was fully opened by stenting without new neurological deficits. Stenting of the basilar artery via a transfemoral route is not always possible even with newer generation stents if the vertebral artery has elongated tortuous curves. Combined surgery and endovascular stenting is one of the alternatives in such cases including our case. PMID- 11482704 TI - Obstruction of Magendie's and Luschka's foramina. Cine-MRI, aetiology and pathogenesis. AB - BACKGROUND: Obstruction of the foramina of Magendie and Luschka represents one possible aetiology for a progressive tetra-ventricular hydrocephalus. METHOD: Our case report is a 58-year-old woman initially presenting with a pseudo vertebrobasilar insufficiency clinical syndrome. A ventriculo-cisternostomy of the third ventricle was performed by an endoscopic procedure with a clinical and radiological success (three year follow-up). FINDINGS: Comparative studies of cerebrospinal fluid (CSF) flow measurements by cine magnetic resonance imaging (cine-MRI) showed that Magendie's and Luschka's foramina permeabilities were restored after the ventriculo-cisternostomy. INRTERPRETATION: This case shows that endoscopic ventriculo-cisternostomy of the third ventricle can provide an effective treatment for specific tetra-ventricular hydrocephalus. Cine-MRI findings open the discussion on possible aetiologies and pathophysiologies. PMID- 11482705 TI - Cranial neuroendocrine carcinoma primarily diagnosed as malignant paraganglioma with rapid progress to a fatal outcome. PMID- 11482706 TI - 3D coronary reconstruction from routine single-plane coronary angiograms: clinical validation and quantitative analysis of the right coronary artery in 100 patients. AB - BACKGROUND: Current coronary angiographic techniques display complex three dimensional (3D) coronary structures in two dimensions (2D). We have developed a 3D reconstruction (3DR) algorithm using standard single-plane angiographic images that allows for 3D display of coronary structures. The purpose of this study was to validate our 3DR algorithm and quantify anatomic characteristics of the right coronary artery (RCA) in vivo. METHODS: Accuracy and reproducibility studies were performed using 3DRs of a coronary phantom and in vivo following 3DRs in 40 patients. The anatomic features of the RCA were then quantified in 100 patients. RESULTS: Comparison of length and bifurcation angles (BA) from the phantom to the 3DRs revealed good accuracy and correlation for both (r = 0.95 and 0.93 respectively), with diameter error of < 7%. In vivo, the average root mean square (RMS) error in the spatial coordinates of the vessel centerlines was 3.12 +/- 0.77 and 3.16 +/- 0.75 mm in 20 left coronary arteries (LCA) and 20 RCAs respectively. Interobserver average RMS error was 3.47 +/- 1.96 mm and intraobserver average RMS error was 3.02 +/- 1.07 and 3.44 +/- 1.57 mm for two different operators (p = NS). The average RCA length was 10.2 +/- 1.7 cm, average radius of curvature (ROC) was 52 +/- 9 degrees, and the average 3D bifurcation angle of the posterior descending artery (PDA) from the RCA was 55 +/- 22 degrees. Foreshortening (FS) of the segments of the RCA in three 'standard' projections ranged from 0-60, 0-75, and 0-82% respectively. CONCLUSIONS: Using our 3DR algorithm patient-specific anatomic characteristics can be accurately displayed and quantified, expanding the information that can be derived from routine coronary angiography. PMID- 11482707 TI - Is the time of 3D representation of the coronary system by X-ray angiography for interventional cardiology almost there? PMID- 11482708 TI - Diagnostic value of resting tricuspid regurgitation velocity and right ventricular ejection flow parameters for the detection of exercise induced pulmonary arterial hypertension. AB - Our objectives were to evaluate resting tricuspid regurgitation velocity (TRV) and right ventricular outflow tract velocity curve (RVOTvc) profiles as markers for development of exercise induced pulmonary arterial hypertension (ExPHT). ExPHT is an elusive cause of dyspnea and fatigue. When present, Doppler echocardiography can detect and quantify elevated pulmonary pressure. However, the characteristics and diagnostic value of resting TRV and RVOTvc indices in patients with ExPHT have not been fully addressed. The study population consisted of 52 subjects (mean age 40.5 +/- 10.9, range 22-68 years) and was divided into three subsets as follows: 1. Patients (n = 22) with overt pulmonary hypertension (PHT), 2. Patients (n = 8) with ExPHT, 3. Healthy, asymptomatic volunteers (n = 22). RVOTvc indices included: Mean and peak velocity, systolic velocity time integral (VTI); velocity time integral at peak velocity (VTImax), acceleration time; ejection time. TRV was used as an index of pulmonary artery systolic pressure. There were significant differences between normals and ExPHT for TRV, acceleration time, VTI(Vmax). TRV and VTImax were predictive of EXPHT in a logistic regression model. CONCLUSION: (1) Patients with ExPHT have distinct Doppler velocity patterns suggesting the presence of a compromised pulmonary vascular bed even with normal pulmonary pressure at rest. (2) TRV and RVOTvc indices have potential diagnostic value in the early detection of ExPHT. PMID- 11482709 TI - The diagnostic value of Doppler echocardiographic indexes of diastolic filling for detecting demand ischemia in anesthetized patients. AB - In awake patients, Doppler indexes of transmitral flow velocities have been shown to be more sensitive for detection of myocardial ischemia than echocardiographic evaluation of systolic wall-motion. The diagnostic value of these indexes in anesthetized patients is unknown. It might differ from that in awake patients because anesthetics and sympathomimetic drugs, which are commonly used in surgical patients with coronary artery disease (CAD), independently affect transmitral flow velocities. METHODS: Several previously published transmitral Doppler echocardiographic indicators of ischemia (marked decreases in the ratio of peak early [E] to peak atrial [A] filling velocity [E/A], in the ratio of early to atrial time-velocity integral [E(I)/A(I)], in E, in E acceleration, and in total diastolic time-velocity integral) were compared with standard wall motion analysis and ST-segment analysis during dobutamine stress echocardiography (DSE) in 17 anesthetized patients with CAD and 7 age-matched control patients at low risk of CAD. RESULTS: All patients with CAD but no control patients showed new systolic wall-motion abnormalities and/or ST-segment changes. Decreases of >10% in E/A and E(I)/A(I) were found in 88% and 71% of the patients with CAD and in 71% and 100% of the control patients, respectively. These decreases were found during dobutamine infusion at as low as 10 mcg/kg/min in several control patients. None of the other studied Doppler indexes were found useful to detect ischemia. CONCLUSIONS: Doppler echocardiography of diastolic transmitral flow velocities is of no appreciable diagnostic value for detecting ischemia in anesthetized patients who receive dobutamine at 10-40 mcg/kg/min. PMID- 11482710 TI - Good correlation between gated single photon emission computed myocardial tomography and contrast ventriculography in the assessment of global and regional left ventricular function. AB - PURPOSE: The purpose of this study was to determine the reliability of the measured left ventricular ejection fraction (LVEF) and wall motion analysis by the recently introduced quantitative electrocardiographically (ECG)-gated myocardial perfusion single-photon emission computed myocardial tomography technique (gated SPECT) (QGS). MATERIALS AND METHODS: We compared technetium-99 m tetrofosmin gated SPECT imaging and contrast ventriculography in the assessment of global and regional left ventricular function in 74 patients with undiagnosed chest pain of whom 27 sustained a previous myocardial infarction. RESULTS: Linear regression analysis demonstrated that gated SPECT determined LVEF correlated well with LVEF determined from contrast ventriculography (y = 0.95x + 1.9, r2 = 0.84, p < 0.0001). Bland-Altman plot analysis showed no systematic difference between the two sets of values derived from the two imaging approaches over a wide range of LVEF values. Exact agreement of segmental wall motion scores was 460 of 518 (89%) segments with a kappa value of 0.76 (p < 0.0001). CONCLUSION: We conclude that gated SPECT imaging is an accurate and reliable clinical tool to accurately measure global and regional left ventricular function. PMID- 11482711 TI - Attenuation smear: a 'paradoxical' increase in counts due to attenuation artifact. AB - BACKGROUND: Attenuation is a well recognized cause of reconstruction artifacts in SPECT imaging. Occasionally, we have noted an increase in activity extending from the apical septal portion of the ventricle in women with significant breast attenuation. Although the idea that attenuation can produce an increase in activity on the reconstructed images seems paradoxical at first, it is consistent with the process of filtered back projection. METHODS: We filled a cardiac phantom with 1 mCi of Technetium-99m, placed it in a water filled anthropomorphic torso phantom and imaged it over a 180 degree orbit. Next, a breast phantom designed to simulate a significant degree of breast attenuation was placed on the torso phantom and imaging was repeated. The images were reconstructed first using conventional filtered back projection then with maximum likelihood. RESULTS: When the phantoms with and without breast attenuation were reconstructed using filtered back projection and compared, the phantom with breast attenuation had a large 'smear' of activity extending anteriorly from the apical septal wall which was very similar to the abnormalities previously noted in clinical images; the phantom without breast attenuation had no such defect. This artifact was significantly less prominent when the images were reconstructed using the maximum likelihood technique. CONCLUSIONS: Attenuation artifact can also produce a seemingly paradoxical increase in counts on the reconstructed image but this phenomenon is consistent with the workings of filtered back projection. PMID- 11482712 TI - Phase-contrast cine MR angiography detection of thoracic aortic dissection. AB - PURPOSE: To assess prospectively the accuracy of phase-contrast cine MR angiography in the detection of thoracic aortic dissection with operative correlation. MATERIALS AND METHODS: One hundred and ninety-seven symptomatic patients suspected of having thoracic aortic dissection or aneurysm as well as 13 patients suspected of having thoracic aortic coarctation and 20 asymptomatic normals (as controls) were examined prospectively with phase-contrast cine MR angiography on a 1.5-T MR imager. Seventy-eight of these patients had operative correlation, and only these 78 patients were included in the statistical analysis. RESULTS: There were 51 true positive and 27 true negative findings of thoracic aortic dissection in this study for an accuracy of 100%. CONCLUSION: Phase-contrast cine MR angiography is an accurate non-invasive imaging technique for evaluating patients suspected of having thoracic aortic dissection. PMID- 11482713 TI - Dobutamine-induced increase of right ventricular contractility without increased stroke volume in adolescent patients with transposition of the great arteries: evaluation with magnetic resonance imaging. AB - OBJECTIVE: Prognosis in patients with surgically corrected (Senning or Mustard) transposition of the great arteries (TGA) depends mainly on right ventricular (RV) function and RV functional reserve. We examined the role of dobutamine stress in the early detection of RV dysfunction in asymptomatic or slightly symptomatic patients with TGA using magnetic resonance imaging (MRI). DESIGN AND PATIENTS: Twelve asymptomatic or slightly symptomatic patients with chronic RV pressure overload, surgically corrected (Mustard or Senning) TGA (age 22.8 (+/- 3.4) years; New York Heart Association (NYHA) class I/II) and nine age matched healthy volunteers (age 27.3 (+/- 4.4) years) were included. MRI was applied both at baseline and during dobutamine stress (start dose 5 microg/kg/min to maximum dose 15 microg/kg/min) to determine RV and left ventricular (LV) stroke volumes (SV) and ejection fraction (EF). RESULTS: At baseline only RVEF was significantly higher in controls than in patients (71 (+/- 9) vs. 57 (+/- 10)%, p < 0.001), other RV parameters were not significantly different between the two examined groups: RVSV (86 (+/- 21) vs. 72 (+/- 27) ml, p = ns), RV end-diastolic volume (EDV) (123 (+/- 37) vs. 123 (+/- 33) ml, p = ns), and heart rate (61 (+/- 10) vs. 69 (+/- 14) bpm, p = ns), respectively. During dobutamine stress RVEF increased significantly both in controls and patients (20 (+/- 16) vs. 17 (+/- 18)%, p < 0.01 and p < 0.02 vs. rest, respectively), but stress RVEF was significantly higher in controls than in patients (85 (+/- 3) vs. 66 (+/- 7)%, p < 0.0001). RVSV increased significantly in controls (22 (+/- 19)%, p < 0.02), and there was no significant increase in RVSV in patients (-10 (+/- 28)%, p = ns). The controls showed no change in RVEDV (2 (+/- 17)%, p = ns), but in patients a significant decrease in RVEDV (-24 (+/- 15)%, p < 0.001) was observed. Maximal heart rate was significantly higher in patients than in controls (122 (+/- 20) vs. 101 (+/- 14) bpm, p < 0.02). CONCLUSION: In asymptomatic or slightly symptomatic patients with surgically corrected TGA dobutamine had a positive inotropic effect on RV, but the increased contractility was not accompanied by an appropriate increase in SV. Our data suggest inadequate RV filling in this category of patients, possibly due to rigid atrial baffles and compromised atrial function or decreased compliance due to RV hypertrophy. PMID- 11482714 TI - Accuracy of MRI evaluation of pulmonary blood supply in patients with complex pulmonary stenosis or atresia. PMID- 11482715 TI - Comparison between segmental wall motion and wall thickening in patients with coronary artery disease using quantitative gated SPECT software. PMID- 11482716 TI - Control of microorganisms in vitro by calcium hydroxide pastes. AB - AIM: The aim of this study was to determine the influence of vehicles on the antimicrobial efficiency of calcium hydroxide. METHODOLOGY: A total of 588 size 50 sterile absorbent paper points, were immersed in various microbial suspensions for 3 min. The points were then placed on Petri dishes and covered with intracanal dressings containing calcium hydroxide: Ca(OH)2 + saline; Ca(OH)2 + camphorated paramonochlorophenol; Ca(OH)2 + 1% chlorhexidine solution: Ca(OH)2 + 3% sodium lauryl sulphate; Ca(OH)2 + Otosporin. After 1 min, 48 and 72 h and 7 days, 147 absorbent paper cones were removed from contact with the intracanal dressings and individually transported and immersed in 5 mL of Letheen Broth, followed by incubation at 37 degrees C for 48 h. Microbial growth was evaluated by turbidity of the culture medium. A 0.1-mL inoculum obtained from the Letheen Broth was transferred to 5 mL of BHI, and incubated at 37 degrees C for 48 h. Bacterial growth was again evaluated by turbidity of the culture medium. Positive BHI tubes were selected and inocula were spread on the surface of BHI agar and incubated at 37 degrees C for 48 h. Gram staining of the BHI growth and from colonies growing on BHI agar was carried out. RESULTS: An antimicrobial effect occurred after 48 h on the cultures of S. mutans, E. faecalis, S. aureus, P aeruginosa, B. subtilis, C. albicans and a mixed culture, irrespective of the intracanal dressing. CONCLUSIONS: Under the conditions of this study, the various vehicles associated with calcium hydroxide pastes did not influence the time required for microbial inactivation. PMID- 11482717 TI - The structure of dentine in the apical region of human teeth. AB - AIM: To study the structure of the apical region of human teeth with emphasis on dentinal tubules and their branches. METHODOLOGY: This descriptive histological study employed demineralized stained sections for light microscopy, demineralized unstained sections for scanning electron microscopy, and undemineralized, acid etched specimens for confocal tandem scanning microscopy. RESULTS: The apical portion of human teeth showed marked variations in structure, including accessory root canals, areas of resorption and repaired resorptions, occasional attached, embedded and free pulp stones, varied amounts of irregular secondary dentine, and even cementum-like tissue lining the apical root canal wall. The apex often deviated from the long axis of the root canal. Primary dentinal tubules were irregular in direction and density. Some areas were devoid of tubules. CONCLUSIONS: The irregular and variable structure of the apical region of human teeth represent special challenges during endodontic therapy. Obturation techniques based on the penetration of adhesives into dentinal tubules are unlikely to be successful and adhesive techniques must depend on impregnation of a hybrid layer. PMID- 11482718 TI - Apical extrusion of debris and irrigants using two hand and three engine-driven instrumentation techniques. AB - AIM: To evaluate the weight of debris and irrigant volume extruded apically from extracted teeth in vitro after endodontic instrumentation using the balanced force technique, a hybrid hand instrumentation technique, and three engine-driven techniques utilizing nickel-titanium instruments (ProFile .04, Quantec 2000 and Pow-R). METHODOLOGY: Five groups of 20 extracted human teeth with single canals were instrumented using one or other of five techniques: balanced force, hybrid, Quantec 2000, ProFile .04, or Pow-R. Debris extruded from the apical foramen during instrumentation were collected into preweighed 1.5 mL tubes. Following instrumentation, the volume of extruded irrigant fluid was determined by visual comparison to control centrifuge tubes filled with 0.25 mL increments of distilled water. The weight of dry extruded dentine debris was also established. RESULTS: Overall, the engine-driven techniques extruded less debris than the manual ones. However, there was no statistical difference between the balanced force technique and the engine-driven methods. The volume of irrigant extruded through the apex was directly associated with the weight of extruded debris, except within the ProFile group. The hybrid technique was associated with the greatest extrusion of both debris and irrigant. CONCLUSIONS: Overall, the engine driven nickel-titanium systems were associated with less apical extrusion. PMID- 11482719 TI - Root and canal morphology of Burmese mandibular molars. AB - AIM: To study the root canal morphology of Burmese mandibular molars using a canal staining and tooth clearing technique. METHODOLOGY: Mandibular molars (331) were collected from indigenous Burmese patients and designated; first (139), second (134), third (58) molars. Following pulp tissue removal and staining of the canal systems with Indian ink, the teeth were decalcified and rendered clear with methyl salicylate. Under magnification (x3), the following features were evaluated: (i) root number and morphology, (ii) number of canals per root, (iii) root canal configuration (Vertucci's classification), (iv) number of apical foramina per root, (v) number and location of lateral canals and (vi) the presence of intercanal communications. RESULTS: Most of the mandibular molars had two separate roots (90% in first molars, 58% in second molars, 53% in third molars) and three-rooted teeth were (10%) confined to first molars. C-shaped roots occurred in 22.4% of mandibular second molars and a further 14.9% had two fused roots. The majority (81-100%) of conical distal roots possessed a simple type I (single canal) configuration. Whilst the canal system of mesial roots was more complex: 52-85% contained two canals, of which type II (two orifices, one foramen) and type IV (two separate canals) were the most prevalent. A broad range of 6.5-70% had intercanal communications. Fused/single-rooted molars had a wide variety of canal system types but intercanal communications were rare except in C shaped roots (33%) of second molars. The majority of roots of all molars contained one or two apical foramina (91-96%) and the apical third had the highest prevalence of lateral canals. CONCLUSIONS: There was a high prevalence of three-rooted mandibular first molars and C-shaped roots/canals in mandibular second molars from a Burmese population. Conical roots tend to have simple canal systems, whilst flatter/broader roots have more complex canal systems. PMID- 11482720 TI - Methodological considerations in the determination of working length. AB - AIM: The aim of this study was to evaluate the diagnostic efficacy of an electronic system for the determination of working length, in comparison with two radiological methods (conventional film and digital radiography). METHODOLOGY: The study sample consisted of 28 root canals belonging to 20 human mandibular teeth. A comparison was made between the working length measurements obtained by two radiological methods (conventional film and digital radiography) and an apex locator, using as gold standard the observation of the file position within the root following selective grinding of the root tissue. RESULTS: The electronic method was satisfactory in 67.8% of cases, versus 50.6% and 61.4% for the conventional and digital radiological methods. respectively. No statistically significant differences occurred between the techniques according to the chi squared and Kruskal-Wallis nonparametric tests. CONCLUSIONS: None of the techniques was totally satisfactory in establishing the true working length. There were no differences between the techniques investigated. PMID- 11482721 TI - The histological effects of four endodontic sealers implanted in the oral mucosa: submucous injection versus implant in polyethylene tubes. AB - AIM: The aim of this study was to evaluate pigmentation and tissue response to four endodontic sealers placed in the oral mucosa of rabbits by either submucous injection or implant in polyethylene tubes. METHODOLOGY: Thirty white New Zealand rabbits were divided randomly into two groups of eight for N-Rickert and AH-26, and two groups of seven for Fillcanal and Sealer 2 6. On the right side of the filter, corresponding to the gingivo-labial sulcus in humans, the sealer was injected; on the left side the sealer was placed within a polyethylene tube and implanted. Direct clinical observations were made at 30, 60 and 90 days. The animals were then sacrificed for histological analysis. RESULTS: After 60 days of observation N-Rickert and AH-26 produced tattoos that became larger by 90 days. Submucous injection produced larger and more numerous pigmentation, when compared to implant in polyethylene tubes. N-Rickert sealer displayed larger and more numerous tattoos when compared to AH-26. Histological analysis showed no differences between the two methods of implantation. All sealers elicited some kind of inflammatory response; the most irritant was Fillcanal, followed by N Rickert and AH-26. Sealer 26 elicited a mild reaction only. CONCLUSIONS: Under the conditions of this study there was no relationship between the method of implantation and the tissue response; the silver-containing sealers produced pigmentation, and the concentration of silver influenced the quantity and size of the tattoos. The sealers elicited various responses when in direct contact with the surrounding tissues: the calcium hydroxide-containing sealer had enhanced healing when compared to the other sealers. PMID- 11482722 TI - Cyclic fatigue of ProFile rotary instruments after prolonged clinical use. AB - AIM: The purpose of the present study was to evaluate resistance to cyclic fatigue of new and used ProFile Ni-Ti rotary instruments. METHODOLOGY: Used instruments were operated in 10 clinical cases using passive instrumentation and a crown-down preparation technique. Cyclic fatigue testing of new and used engine driven instruments was then performed with a specific device which allowed the instruments to rotate freely inside a stainless steel artificial canal, whilst maintaining conditions close to the clinical situation. Instruments were rotated until fracture occurred and time to fracture was visually recorded with a chronometer. RESULTS: A significant reduction of rotation time to breakage (life span) was noted between new and used instruments. In all sizes new instruments were significantly more resistant than used ones (two-sample t-test, P < 0.01). No instrument underwent intracanal failure during clinical use. CONCLUSIONS: Prolonged clinical use of Ni-Ti engine-driven instruments significantly reduced their cyclic fatigue resistance. Nevertheless, each rotary instrument was successfully operated in up to 10 clinical cases without any intracanal failure. PMID- 11482723 TI - A methodology for quantitative evaluation of root canal instrumentation using microcomputed tomography. AB - AIM: The aim of this paper was to present an objective methodology for quantitative evaluation of root canal instrumentation using microcomputer tomography, together with developed software based on a constructed mathematical model. METHODOLOGY: A desktop X-ray micro-CT scanner (SkyScan 1072) was used to provide data sets of an extracted mandibular molar before and after instrumentation (ProFile 0.04 Taper instruments, Dentsply Maillefer, Ballaigues, Switzerland) that were stored for later use by software. A volume visualization package (T3D) was applied to obtain 3D renderings of the molar to illustrate the qualitative visualization capacity. Next, medical image volume fusion software was used to allow alignment of pre and post image volumes. Finally, software was developed to make quantitative measurements and to provide additional qualitative information on the registered image volumes. This procedure implemented a true 3D mathematical model for quantification of instrumentation effects. Using a local co-ordinate frame, perpendicular reslices were made at five different levels within the mesio-buccal canal to evaluate transportation and centring ability. RESULTS: At first accurate and detailed 3D renderings were obtained. The general and local canal shape before and after preparation could visually be examined in 360 degrees rotation. Numerical values were obtained for volume, volume changes and transportation. Centring ability was calculated by centring ratio and centre movement. CONCLUSIONS: This methodology is a new and objective way for quantitative evaluation of root canal instrumentation using microcomputer tomography and dedicated software. PMID- 11482724 TI - Enterococcus faecalis--a mechanism for its role in endodontic failure. AB - AIM: The aim of this study was to identify a possible mechanism that would explain how E. faecalis could survive and grow within dentinal tubules and reinfect an obturated root canal. METHODOLOGY: Cells of Streptococcus gordonii DL1, Streptococcus mutans NG8, or E. faecalis JH2-2 were grown in brain heart infusion broth containing various amounts of human serum for 56 days. The ability of the three species to invade dentine and bind to immobilized type I collagen in the presence of human serum was assessed by dentine invasion and microtitre well experiments. RESULTS: All three species remained viable over the period of the experiment when grown in human serum. Cells of all three bacteria were able to invade dentine and bind to immobilized collagen. Both of these properties were inhibited by the presence of collagen in the cell solution. Human serum inhibited dentine invasion and collagen adhesion by S. gordonii DL1 and S. mutans NG8, whilst dentine invasion by E. faecalis JH2-2 was reduced in the presence of serum, but not inhibited, and binding to collagen was enhanced. CONCLUSIONS: It is postulated that a virulence factor of E. faecalis in failed endodontically treated teeth may be related to the ability of E. faecalis cells to maintain the capability to invade dentinal tubules and adhere to collagen in the presence of human serum. PMID- 11482725 TI - The microscopic diagnosis and treatment of a mandibular second premolar with four canals. AB - AIM: The purpose of this article is to report the case of a mandibular second premolar with four distinct canals. SUMMARY: It is generally recognized that incomplete instrumentation and cleaning of root canals will lead to failure in root-treated teeth. Before root canal treatment is performed, the clinician should be aware of the configuration of the pulp space of the tooth that is to be treated. Frequently, root canals are left untreated because the clinician fails to identify their presence, particularly in teeth that have anatomical variations or additional root canals. Traditionally, additional canals have been detected by clinical examination of the floor of the pulp chamber, and sometimes found radiographically. More recently microscopic examination of the pulp chamber using endodontic microscopy has assisted the identification of extra canals. A case report is presented that deals with the successful treatment of a mandibular second premolar with four canals. KEY LEARNING POINTS: Even in teeth with a low frequency of abnormal root canal anatomy, the possibility of additional root canals has to be considered in the clinical and radiographic examination of the patient. The ability to recognize and locate unsuspected canals is enhanced by microscopic investigation of the pulp chamber. PMID- 11482726 TI - The standardized-taper root canal preparation--Parts 1 to 6. PMID- 11482728 TI - Sensitive and simple determination of mannitol in human brain tissues by gas chromatography-mass spectrometry. AB - A simple, reliable and sensitive gas chromatographic-mass spectrometric method was devised to determine the level of mannitol in various human brain tissues obtained at autopsy. Mannitol was extracted with 10% trichloroacetic acid solution which effectively precipitated brain tissues. The supernatant was washed with tert.-butyl methyl ether to remove other organic compounds and to neutralize the aqueous solution. Mannitol was then derivatized with 1-butaneboronic acid and subjected to GC-MS. Erythritol was used as an internal standard. For quantitation, selected ion monitoring with m/z 127 and 253 for mannitol and m/z 127 for internal standard were used. Calibration curves were linear in concentration range from 0.2 to 20 microg/0.1 g and correlation coefficients exceeded 0.99. The lower detection limit of mannitol in distilled water was 1 ng/0.1 g. Mannitol was detected in control brain tissues, as a biological compound, at a level of 50 ng/0.1 g. The precision of this method was examined with use of two different concentrations, 2 and 20 microg/0.1 g, and the relative standard deviation ranged from 0.8 to 8.3%. We used this method to determine mannitol in brain tissues from an autopsied individual who had been clinically diagnosed as being brain dead. Cardiac arrest occurred 4 days later. PMID- 11482729 TI - Molecular heterogeneity of amyloid beta2-microglobulin and modification with advanced glycation end products. AB - By using liquid chromatography-electrospray ionization mass spectrometry, Western blotting and N-terminal amino acid sequence analysis, we characterized the molecular heterogeneity and advanced glycation end product (AGE) modification of beta2-microglobulin (beta2m) extracted from the amyloid tissue of a hemodialysis patient. Amyloid beta2m was composed of full-length beta2m, truncated beta2m and dimer beta2m. Truncated beta2m and dimer beta2m were modified with AGEs such as imidazolone and N(e)-(carboxymethyl)lysine, and showed fluorescence characteristic of AGE. Truncated beta2m species were formed by cleavage between amino acid residues of Pro6/Ile7, Gln/Val9 and Val9/Tyr10. Heterogeneous dimer beta2m species showed the molecular masses of 22,591 and 22 675, which resulted from cross-linking between truncated beta2m. PMID- 11482730 TI - Sensitive determination of pethidine in body fluids by surface ionization organic mass spectrometry. AB - We have presented a simple and sensitive method for determining pethidine, a narcotic analgesic drug in body fluids by gas chromatography (GC)/surface ionization organic mass spectrometry (SIOMS). Good linearity was obtained in the range of 0.625-25 ng/ml of whole blood and urine by mass chromatography, and in the range of 0.05-2 ng/ml of whole blood by selected ion monitoring (SIM). Pethidine and diphenylpyraline (internal standard) were extracted from body fluids with Bond Elut Certify cartridges; their recoveries were above 95%. The detection limits (signal-to-noise ratio=3) were estimated to be 0.2 ng/ml of whole blood or urine by mass chromatography, 0.02 ng/ml of whole blood by SIM. PMID- 11482731 TI - Determination of acrolein by headspace solid-phase microextraction gas chromatography and mass spectrometry. AB - We developed a headspace solid-phase microextraction (headspace SPME) method to measure acrolein in human urine. This new technique resolves some problems with the headspace gas chromatography and mass spectrometry (GC-MS) method which we developed previously. With the original method, a column and a filament were damaged by the injection of air. A 0.5-ml urine (or phosphate-buffered saline) sample in a glass vial containing propionaldehyde as an internal standard was heated for 5 min. The SPME fiber (65 microm carbonwax-divinylbenzene fiber) was exposed to the headspace and then inserted into a GC-MS instrument in which a DB WAX capillary column (30 m x 0.32 mm, film thickness 0.5 degrees m) was installed. The total analysis time was 15 min. The inter-assay and intra-assay coefficients of variation were 10.07 and 5.79%, respectively. The calibration curve demonstrated good linearity throughout concentrations ranging from 1 to 10,000 nM. The headspace SPME method exhibits high sensitivity and requires a short analysis time as well as the previous method. We conclude that this method is useful to measure urinary acrolein. PMID- 11482732 TI - Clinical applications of tandem mass spectrometry: ten years of diagnosis and screening for inherited metabolic diseases. AB - This paper reviews the clinical applications of tandem mass spectrometry (MS-MS) in diagnosis and screening for inherited metabolic diseases in the last 10 years. The broad-spectrum of diseases covered, specificity, ease of sample preparation, and high throughput provided by the MS-MS technology has led to the development of multi-disorder newborn screening programs in many countries for amino acid disorders, organic acidemias, and fatty acid oxidation defects. Issues related to sample acquisition, sample preparation, quantification of metabolites, and validation are discussed. Our current experience with the technique in screening is presented. The application of MS-MS in selective screening has revolutionized the field and made a major impact on the detection of certain disease classes such as the fatty acid oxidation defects. New specific and rapid MS-MS and LC-MS MS methods for highly polar small molecules are supplementing or replacing some of the classical GC-MS methods for a multitude of metabolites and disorders. New exciting applications are appearing in fields of prenatal, postnatal, and even postmortem diagnosis. Examples for pitfalls in the technique are also presented. PMID- 11482733 TI - Diagnosis of inborn errors of metabolism using filter paper urine, urease treatment, isotope dilution and gas chromatography-mass spectrometry. AB - This review will be concerned primarily with a practical yet comprehensive diagnostic procedure for the diagnosis or even mass screening of a variety of metabolic disorders. This rapid, highly sensitive procedure offers possibilities for clinical chemistry laboratories to extend their diagnostic capacity to new areas of metabolic disorders. The diagnostic procedure consists of the use of urine or filter paper urine, preincubation of urine with urease, stable isotope dilution, and gas chromatography-mass spectrometry. Sample preparation from urine or filter paper urine, creatinine determination, stable isotope-labeled compounds used, and GC-MS measurement conditions are described. Not only organic acids or polar ones but also amino acids, sugars, polyols, purines, pyrimidines and other compounds are simultaneously analyzed and quantified. In this review, a pilot study for screening of 22 target diseases in newborns we are conducting in Japan is described. A neonate with presymptomatic propionic acidemia was detected among 10,000 neonates in the pilot study. The metabolic profiles of patients with ornithine carbamoyl transferase deficiency, fructose-1,6-bisphosphatase deficiency or succinic semialdehyde dehydrogenase deficiency obtained by this method are presented as examples. They were compared to those obtained by the conventional solvent extraction methods or by the tandem mass spectrometric method currently done with dried filter blood spots. The highly sensitive, specific and comprehensive features of our procedure are also demonstrated by its use in establishing the chemical diagnosis of pyrimidine degradation defects in order to prevent side effects of pyrimidine analogs such as 5-flurouracil, and the differential diagnosis of three types of homocystinuria, orotic aciduria, uraciluria and other urea cycle disorders. Evaluation of the effects of liver transplantation or nutritional conditions such as folate deficiency in patients with inborn errors of metabolism is also described. PMID- 11482734 TI - Initial catabolic steps of isoleucine, the R-pathway and the origin of alloisoleucine. AB - The initial catabolic steps of isoleucine by mammals has been misunderstood and misapprehended in the scientific literature for many years. The suggestion that the interconversion of isoleucine and alloisoleucine occurs through the keto-enol racemization of their respective transaminated alpha-keto acids was first tentatively advanced by Alton Meister in the early 1950s, and accepted without hard confirming evidence by many authors. It will be shown in this brief review that isoleucine is converted to alloisoleucine with conservation of a 15N label denying the intermediacy of the alpha-keto acids, and that alloisoleucine arises as an unavoidable consequence of isoleucine transamination. PMID- 11482735 TI - Neonatal ketosis is not rare: experience of neonatal screening using gas chromatography-mass spectrometry. AB - The causes and effects of transient neonatal ketosis, discovered during a pilot study of screening for abnormalities in neonatal metabolism using gas chromatography-mass spectrometry, were investigated. Of the 21,342 neonates that were screened, 47 had significant ketosis. The organic acid profile accompanying ketosis in the urine of neonates followed the pattern of ketotic dicarboxylic aciduria in approximately half of the cases. Ketosis was more often found in neonates nourished by breast feeding (33 out of 47). Over half of the neonates showing ketosis (28 out of 47) were asymptomatic. When normal neonates and neonates testing positive for ketosis were compared, no statistically significant correlations were found with regard to birth mass, gestational period, or gender. However, neonates with ketosis tended to have low mass gain rates in the 5 days from birth and a statistically significant difference was found in this regard in comparison to normal neonates (P<0.0001). From the above results, development of ketosis in neonates was found to be possible even in normal subjects. Most ketosis in neonates was also found to depend largely on nourishment after birth. Existence of an asymptomatic ketosis category was also suggested. PMID- 11482736 TI - Simple gas chromatographic-mass spectrometric procedure for diagnosing pyrimidine degradation defects for prevention of severe anticancer side effects. AB - Inborn errors of pyrimidine degradation, dihydropyrimidine dehydrogenase deficiency and dihydropyrimidinase deficiency, are less rare than has generally been assumed. Many asymptomatic cases have been reported, and in patients with symptoms, the clinical abnormalities are variable and nonspecific. Withdrawal of pyrimidine analogues such as 5-fluorouracil (5FU), a commonly used anticancer drug, from the cancer chemotherapy regimens of patients with pyrimidine degradation deficiencies, however, is critical because 5FU is degraded in vivo by pyrimidine-degradative enzymes. Patients with these deficiencies suffer from severe neurotoxicity, sometimes leading to death, following administration of 5FU, and even otherwise asymptomatic homozygotes or heterozygotes may develop severe clinical symptoms upon administration of such medication. Therefore, a rapid and specific method for identifying cancer patients with these enzyme deficiencies prior to treatment with 5FU is critical. To address this problem, we established methods for highly sensitive yet specific determinations of thymine, uracil, dihydrothymine, dihydrouracil, orotate and creatinine simultaneously in 0.1-ml liquid urine or filter-paper urine. This method involves stable isotope dilution, a simplified urease treatment previously described and gas chromatography-mass spectrometry without prior fractionation. The high recovery and low C.V. values were obtained and healthy control values were also determined for these metabolites. Using artificially prepared urine specimens simulating these disorders. the chemical diagnosis can be made clearly, and no further analysis appears to be required for differential chemical diagnosis. PMID- 11482737 TI - Screening for inborn errors of metabolism using gas chromatography-mass spectrometry. AB - Screening of newborns for inborn errors of metabolism (IEM) in China is both a challenging and undeveloped area for gynecologists and pediatricians. Since 1999, the Capital Institute of Pediatrics has been studied as regards screening for IEM using advanced gas chromatography-mass spectrometry (GC-MS) method in collaboration with the Matsumoto Institute of Life Science (MILS), Japan, and has successfully diagnosed 51 cases of IEM in a total of 393 patients. Galactosemia, phenylketonuria and methylmalonic acidemia were the most frequent disorders among 51 cases of IEM. Treatment by suitable drugs and/or diet therapy was very effective in the most cases. PMID- 11482738 TI - Urinary organic acids in peroxisomal disorders: a simple screening method. AB - Using GC-MS, we studied urinary organic acids in 20 Japanese patients with peroxisomal disorders, including Zellweger syndrome (ZS), neonatal adrenoleukodystrophy, and single deficiency of peroxisomal beta-oxidation enzymes. Non-ketotic dicarboxylic aciduria with elevated sebacate/adipate molar ratio was observed in 19 of the 20 patients. Elevation of 2-hydroxysebacate and epoxydicarboxylic acids were seen in 13 and 18, respectively. Tyrosyluria was remarkable in all patients. In two ZS patients, we tracked the time course from birth to infancy, and all the above stated findings were detected, except for one sample. Urinary organic acid analysis is indeed useful for screening subjects with peroxisomal disorders. PMID- 11482739 TI - Gas chromatographic-mass spectrometric screening for organic acidemias using dried urine filter paper: determination of alpha-ketoacids. AB - There are several organic acid disorders that require information on alpha ketoacids, such as maple syrup urine disease or alpha-ketoadipic acidemia. The recovery, stability and diagnostic availability of alpha-ketoacids in dried urine filter paper analyzed by GC-MS with oxime-trimethylsilyl derivatization was studied for organic acidemia screening. The recovery of all nine types of alpha ketoacids tested, but for phenylpyruvate, 2-ketoadipate, and p-OH-phenylpyruvate, from filter paper samples was acceptable. The stability of pyruvate, branched chain alpha-ketoacids, alpha-ketoadipate and alpha-ketoglutarate was stable for at least 28 days, although some alpha-ketoacids such as succinylacetone were unstable. It indicated it was difficult to diagnose only tyrosinemia type 1 among nine specimens from organic acidemia patients tested. The method could be applied to global organic acidemia screening. PMID- 11482740 TI - New method of derivatization and headspace solid-phase microextraction for gas chromatographic-mass spectrometric analysis of amphetamines in hair. AB - A simple method for hair analysis of methamphetamine (MAMP) and amphetamine (AMP) by gas chromatography-mass spectrometry (GC-MS) was developed using simultaneous headspace solid-phase microextraction (HS-SPME) with derivatization. After alkaline-digestion of hair, the analytes derivatized with heptafluoro-n-butyryl chloride were adsorbed on a polydimethylsiloxane-coated fiber by HS-SPME and analyzed by GC-MS. Their mass spectra were, respectively, observable at 1 ng per sample. The standard curves in the range of 0.1-100 ng were linear. The intra-day coefficients of variation at each 0.5 ng were 12.5% for AMP and 3.8% for MAMP. The applicability of this method was demonstrated in some case studies. PMID- 11482741 TI - Can drugs or micronutrients prevent cataract? AB - Cataract is the major cause of blindness and of visual impairment worldwide, so its prevention is of the greatest importance. At present no drug therapy is licensed for use in the UK or the US, so the only treatment for cataract is by surgery, which is expensive and has adverse effects. This article reviews research on prevention of cataract by a variety of agents, including micronutrients as well as drugs. Benefits have been claimed for many compounds or mixtures and this review concentrates on those most extensively studied. Information on possible benefits of putative anticataract agents comes from a variety of approaches, from laboratory experiments, both in vitro and in vivo, to epidemiological studies in patients. Sorbitol-lowering drugs were the first to be examined systematically and progressed to clinical trials which were disappointing, and now the entire rationale for their use in prevention of cataract is questionable. Micronutrients showed little promise in animals but came to clinical trial in patients with cataract without the publication of any major benefit. Pantethine showed more promise in animal studies but the only clinical trial was abandoned early. A variety of laboratory and epidemiological evidence supports the benefits of aspirin-like drugs but there has been no trial specifically in patients with cataract. Add-on studies to trials of aspirin for other indications have not been encouraging. Research into other compounds is interesting but less advanced. PMID- 11482742 TI - Helicobacter pylori-associated peptic ulcer disease in older patients: current management strategies. AB - The incidence of peptic ulcer and its severe complications, i.e. bleeding or perforation, is increasing in elderly patients worldwide. The prevalence of Helicobacter pylori infection in patients with peptic ulcer aged over 65 years has been reported to range from 58 to 78%. However, in elderly patients hospitalised for ulcer disease, the rate of diagnostic screening or treatment for H. pylori infection was less than 60%, and only 50 to 73% of patients who had a positive H. pylori test were treated with antibacterials. The eradication of H. pylori infection is known to be of proven benefit for elderly patients with H. pylori-associated ulcer disease. Significant improvement of the clinical outcome, and reduction of ulcer recurrences, symptoms and histological signs of ulcer associated chronic gastritis activity, as well as decreased costs in elderly healthcare, all result from successful therapy. Proton pump inhibitor (PPI)-based triple therapy regimens including clarithromycin, amoxicillin and/or nitroimidazoles are highly effective and well tolerated in elderly patients, particularly if therapy is of a short duration and low doses of both the PPI and clarithromycin are used. Resistance of H. pylori to antibacterials and low compliance are the major reasons for treatment failure. Surveillance of H. pylori susceptibility to antibacterials at the regional level and enhanced compliance programmes give promising results that suggest new approaches to anti-H. pylori treatment, especially in elderly patients. The role of H. pylori infection in nonsteroidal anti-inflammatory drug (NSAID)-related peptic ulcer still remains controversial. At present, no clear evidence supports the testing and treatment of H. pylori infection for the prevention of drug-related peptic ulcer in elderly patients receiving an NSAID or aspirin (acetylsalicylic acid). After therapy, elderly patients with peptic ulcer may be re-evaluated by invasive methods, i.e. endoscopy and gastric biopsies. or by noninvasive methods. In elderly patients, the 13C-urea breath test demonstrated significantly higher sensitivity, specificity and diagnostic accuracy for detecting H. pylori infection than anti H. pylori antibodies. PMID- 11482743 TI - Orthostatic hypotension in patients with Parkinson's disease: pathophysiology and management. AB - Orthostatic hypotension is common in elderly patients, and is now considered to be an important prognostic factor for cognitive decline and mortality. In patients with Parkinson's disease, the prevalence of symptomatic orthostatic hypotension may be as high as 20%. Two factors could explain this high prevalence. First, dopaminergic drugs may induce or worsen orthostatic hypotension. Secondly, Parkinson's disease is a cause of primary autonomic failure with an involvement of the peripheral autonomic system as shown by the ubiquitous distribution of Lewy bodies and reduced iobenguane [metaiodobenzylguanidine (MIBG)] cardiac uptake. These pathological and pharmacological characteristics clearly differentiate autonomic failure of Parkinson's disease from multiple system atrophy. If autonomic abnormalities appear to be present from the first stage of the disease, early onset (within the first year) of symptomatic orthostatic hypotension in the course of parkinsonism can be considered as an exclusion criteria for idiopathic Parkinson's disease. No specific clinical trials have evaluated the effects of antihypotensive drugs in patients with Parkinson's disease and thus no specific therapeutic strategy can be recommended. The management of orthostatic hypotension in patients with Parkinson's disease should always start with patient education and nonpharmacological treatment. Drug therapy should be reserved for symptomatic patients who do not get benefit from nonpharmacological management. Among the available drugs, alpha1-adrenergic agonists (mainly midodrine) or plasma volume expanders (mainly fludrocortisone) are the most frequently used. There are also some drugs that are currently investigational such as yohimbine and droxidopa. Other drugs such as desmopressin or octreotide may be of interest in some situations. Domperidone is widely used in patients with parkinsonism with no proven effect on orthostatic hypotension. PMID- 11482744 TI - Chronic depression in the elderly: approaches for prevention. AB - Depression in old age frequently follows a chronic and/or relapsing course, related to medical comorbidity, cognitive impairment and depletion of psychosocial resources. As endorsed by the US National Institutes of Health (NIH) Consensus Development Conference on the Diagnosis and Treatment of Late Life Depression, a major goal of treatment is to prevent relapse, recurrence and chronicity. We believe that most, if not all, elderly patients with major depressive episodes are appropriate candidates for maintenance therapy, because of the vulnerability to relapse and recurrence and because of the favourable benefit to risk ratio of available treatments. Antidepressant pharmacotherapy is the mainstay of this therapeutic goal, but psychosocial approaches (especially interpersonal psychotherapy) have also been shown to contribute significantly to prevention of a chronic depressive illness and to prevention of the disability that attends depression. Studies published to date have established the long term or maintenance efficacy of the tricyclic antidepressant nortriptyline. Current, ongoing studies are addressing the maintenance efficacy of paroxetine and citalopram to prolong recovery in depression associated with old age. These studies are focusing particularly on patients aged 70 years and above, who are at high risk of recurrence, and on patients in primary care settings, where under recognition and under-treatment of depression in the elderly have been costly from a public health perspective in terms of increased medical utilisation, burden to patients and families, and high rates of suicide. Depression in old age is a major contributor to the global burden of illness-related disability, but it is extremely treatable if appropriate pharmacotherapy is prescribed and accepted by patients and their caregivers. PMID- 11482745 TI - Economics of antihypertensive therapy in the elderly. AB - Because of the high incidence of morbidity and mortality associated with hypertension in the elderly, the treatment of hypertension in this patient group must involve consideration of clinical, humanistic and economic outcomes. The most frequently used method of pharmacoeconomic analysis for antihypertensive therapy involves cost-effectiveness analysis, although several other methods are available. Current evidence reveals a trend toward cost effectiveness of antihypertensive treatment in elderly patients. However, these formal analyses are limited by the need for extrapolation of data regarding efficacy and level of risk from epidemiological and randomised trials, information which is often lacking. To incorporate economic factors into clinical decision making, other measures of economic impact should be explored. The economic impact of antihypertensive therapy is affected by the level of risk for the patient and the efficacy of the treatment. Data indicate that the risk of morbidity and mortality related to hypertension increases with age and that current antihypertensive drugs reduce this risk. When choosing an antihypertensive agent, the following parameters should be considered: acquisition cost, likelihood of adverse effects and other determinants of treatment adherence, and individual predictors of response. The economic outcomes will be maximised if prudent drug selection is supplemented by appropriate diagnostic and classification procedures and reduction of cardiovascular risk factors other than hypertension. The accumulation of data addressing the risks and benefits of therapy for the very old and the comparative efficacy of newer antihypertensive therapies will further clarify the decision-making process. PMID- 11482747 TI - Twelve-month treatment of overactive bladder: efficacy and tolerability of tolterodine. AB - CONTEXT: Tolterodine is a bladder-selective antimuscarinic agent designed for the treatment of overactive bladder. Traditional antimuscarinic therapies are poorly tolerated due to a high incidence of anticholinergic adverse events and consequently few patients remain on long term therapy. OBJECTIVE: To evaluate the long term efficacy and tolerability of tolterodine in patients with symptoms of overactive bladder. DESIGN: Twelve-month open-label extension of 4 randomised, placebo-controlled, double-blind, multinational, multicentre trials of 4 weeks' duration. PATIENTS: 714 patients (aged 18 to 92 years) with symptoms of overactive bladder who completed the double-blind portion of the studies. INTERVENTION: Tolterodine 2 mg twice daily for up to 12 months. MAIN OUTCOME MEASURES: Micturition diary variables: number of micturitions per 24 hours, number of urge incontinence episodes per 24 hours, mean urine volume voided per micturition. Safety variables: adverse events, study discontinuation rate. RESULTS: A total of 441 patients (62%) completed 12 months' open-label treatment with tolterodine, which significantly reduced the number of micturitions per 24 hours [mean change -2.4, 95% confidence interval (CI) -2.7 to -2.2, median change -20%, p < 0.0001] and number of urge incontinence episodes per 24 hours (mean change -1.3, 95% CI -1.6 to -1.0, median change -74%, p < 0.0001), while the mean volume voided per micturition was significantly increased (+33 ml, 95% CI +28 to +38, median change +18%; p < 0.0001). 41% of patients reported dry mouth (27% mild, 10% moderate, 3% severe). Dosage reduction to 1 mg twice daily was required in 23% of patients. 15% of patients withdrew from the study due to adverse events, with 5% having associated dry mouth. CONCLUSIONS: The high percentage of patients completing 12 months' treatment indicates that tolterodine is an effective and well tolerated agent for long term treatment of overactive bladder. PMID- 11482746 TI - Clinical and economic factors in the treatment of behavioural and psychological symptoms of dementia. AB - The prevalence of behavioural and psychological symptoms of dementia (BPSD) exceeds 50%. They cause distress to patients and caregivers, increase resource utilisation of various kinds, and form a high risk for accelerated psychiatric care through institutionalisation. Although evidence for current pharmacological treatment is not strong and the construct of BPSD is still not very clear, future aspects of treatment of BPSD may be positive. If we look at overall success rates of the antipsychotics, the traditional antipsychotics have the highest combined success rate of 63.1%, whereas the novel antipsychotics have an overall success rate of 56.1%. Haloperidol is the drug with the highest success rate of 65.4%, although this drug is associated with parkinsonian adverse drug reactions. Newer antipsychotics show promise in treating BPSD, but more convincing evidence (e.g. from randomised clinical trials) is required. We provide an overview of the clinical, epidemiological and economic aspects of BPSD and a review of the available literature on their pharmacological treatment. Although only 1 pharmacoeconomic study has been conducted on BPSD, it seems likely that these manifestations drastically increase the burden of dementia. PMID- 11482748 TI - The effects of cognitive impairment and substance abuse on psychiatric hospitalizations. AB - Previous studies have demonstrated a relationship between impairment in executive functioning and hospital and community tenure for people with schizophrenia. However, while it has been clearly established that comorbid substance abuse has a profound negative impact on the functioning of people with schizophrenia, no studies have examined the relative effect of cognitive impairment to substance use in predicting rehospitalization in this population. The present study examined the extent to which impairment on the Wisconsin Card Sorting Test (WCST) and substance abuse are correlated with lifetime psychiatric hospitalizations for outpatients with schizophrenia. Substance abuse was a significant predictor of prior hospitalizations and impairment on the WCST was a significant predictor of the months hospitalized. The findings suggest that both substance abuse and cognitive impairment need to be addressed in order to reduce hospitalizations and time in the hospital. PMID- 11482749 TI - The prediction of mental health service use in residential care. AB - The purpose of this study was to characterize the clinical and psychosocial factors of residents living in psychiatric nursing homes, assess residents' levels of mental health service utilization, and examine the factors that predict the utilization of mental health services. Data were collected from 200 randomly selected residents with schizophrenia living in four intermediate care facilities. Fewer than 60% of residents received mental health services beyond medication and nearly one-half of the residents were readmitted to the hospital in the course of a year. Family contact and involvement in activities were associated with mental health service utilization. Hospital readmission was predicted, not by substance use, but rather by not using substances. There is a growing need among service providers to better identify relevant factors that are important in treatment planning and service delivery. Attention to these issues may impact treatment provision and outcomes for persons with schizophrenia and their families. PMID- 11482750 TI - Quality of life in boarding houses and hostels: a residents' perspective. AB - In the last forty years deinstitutionalization has transferred the care of people with a serious mental illness from the psychiatric hospitals to community based facilities. More recently it has been questioned whether these new facilities offer the anticipated benefits of quality of life. This study examines the Quality of Life (QOL) of people diagnosed with schizophrenia living in two different accommodation facilities, hostels and boarding houses. QOL is examined from the resident's perspective. Lehman's (1988b) QOL Interview was used to measure objective, subjective, and global QOL of 60 participants in three hostels and two boarding house clusters. Hostel and boarding house data were compared and results showed that residents preferred boarding house accommodation. Overall, residents of both accommodation facilities reported satisfaction with QOL, and indicated that they regard them as asylum or sanctuary from the outside world. PMID- 11482751 TI - Perceived needs and service use of Spanish speaking monolingual patients followed at a Hispanic clinic. AB - This study compared Spanish speaking monolingual patients at a Hispanic Clinic, with Hispanics and non-Hispanics at a conventional mental health center. A questionnaire in English and Spanish surveyed sociodemographic characteristics and subjectively reported health needs. The monolingual Hispanic Clinic patients perceived a significantly greater need for help with health and benefits, and better access to physical health care providers. Hispanics seen at the Hispanic Clinic report more difficulties managing medications than the other groups suggesting that this problem is not adequately addressed by receiving services in a culturally sensitive clinic, possibly due to the persistent effect of language barriers and low level of education. Additional interventions may be needed to address this problem. PMID- 11482752 TI - Is stealing a gateway crime? AB - Stealing behaviors are common among youth and adolescents and often begin in early ages due to a variety of developmental, behavioral, and social factors. This study explores the prevalence of stealing behaviors in a South Texas juvenile detention facility. Researchers discovered stealing was the most frequently reported offense followed by the lack of supervision. Youth arrested for stealing were also more likely to be arrested prior to their twelfth birthday, have subsequent charges that may include assault, and have multiple charges. Results with implications for interventions are provided. PMID- 11482753 TI - The incarceration of individuals with severe mental disorders. AB - This study examines the extent to which severely mentally disabled (SMD) patients in one county mental health system were incarcerated in the local jail and examines characteristics of a sample (N = 30) of such individuals. We found that in the study year, 7.9% of known SMD patients had at least one incarceration in the county jail. Diagnoses were predominantly in the schizophrenia spectrum with 70% also actively abusing substances at the time of incarceration. The majority of crimes were non-violent and substance abuse related. Half of the sample was judged to be candidates for diversion programs. Our findings are consistent with recent literature confirming that substance abusing SMD individuals are at high risk of incarceration and could benefit from integrated mental health and substance abuse treatment. PMID- 11482754 TI - Analgesic properties of Epilobium angustifolium, evaluated by the hot plate test and the writhing test. AB - The analgesic properties of Epilobium angustifolium (Ea), a plant containing flavonoids with anti-inflammatory activity, have not been sufficiently studied so far. Thus, we decided to evaluate, by the classical hot plate test and the writhing test, the analgesic effect of a dry extract of Ea obtained by evaporating a commercially available mother tincture. In the former assay, the effect of Ea (380 mg/kg) was slightly lower than that of morphine (10 mg/kg s.c.). In the writhing test, which is more sensitive for non-steroidal analgesics, the effect of Ea was already significant (P < 0.05) at 95 mg/kg while at doses > or = 190 mg/kg, its activity was similar to that of lysine acetylsalicylate (300 mg/kg). The LD50 of this dry extract of Ea was 1.4+/-0.1 g/kg. Further studies are necessary for the identification of the active principles and the elucidation of their mechanism of action. PMID- 11482755 TI - Antimicrobial activity of Epilobium spp. extracts. AB - The antimicrobial activity of the Epilobium angustifolium, E. hirsutum, E. palustre, E. tetragonum and E. rosmarinifolium ethanolic extracts was studied in vitro on Gram-positive and Gram-negative bacteria, yeasts and fungi. The cytotoxicity of the extracts was also evaluated using the Artemia salina test. All the extracts showed antimicrobial activity in a range of concentrations between 10 and 650 microgml of dry extract. E. angustifolium and E. rosmarinifolium had the most broad spectrum of action inhibiting bacteria, yeasts and fungi. The extracts were devoid of toxicity on Artemia salina within the range of antimicrobial concentrations, suggesting that the action is selective on microorganisms. PMID- 11482756 TI - Interactions between A-9THC and capsaicin on isolated lamb bladder detrusor. AB - A number of studies have demonstrated that capsaicin, a capsicum alkaloid, can affect isolated bladder tissue with either a relaxation or a contraction, depending on the species, by acting on VR1 receptors. In a previous work on isolated lamb detrusor, we demonstrated that capsaicin generally produces a relaxation of the tissue; this relaxation seems to be mediated by CGRP. Endogenous cannabinoids, such as anandamide, produce some of their actions by stimulating VR1 receptors and this seems to cause the release of peptides, e.g. CGRP. The aim of this work was to ascertain whether a cannabinoid, delta-9 tetrahydrocannabinol (delta-9THC), was able to interfere with the response of the isolated lamb detrusor to capsaicin. A-9THC, at concentrations between 1.6 x 10( 7) and 1.3 x 10(-6) M, displayed no activity on tissues. Instead, following delta 9THC, most of the tissues responded to capsaicin with a contraction that was abolished by atropine (9.0 x 10(-7) M). It has been reported that cannabinoids can inhibit the release of CGRP by stimulation of CB1 and CB2 cannabinoid receptors. Delta-9THC could act stimulating these receptors and thus inhibiting CGRP release and vesical relaxation. The muscle relaxing component removal could favour the contracting component, usually not active. PMID- 11482757 TI - Studies on vasoconstrictor activity of Curculigo pilosa extracts and of its isolated compounds. AB - The Curculigo pilosa total extract, its butanolic fraction (0.5 microg-100 mg/kg) and the most active in vitro compound structurally similar to adrenaline, pilosidine (10 ng-l mg/kg), caused a reversible and dose-dependent increase in blood pressure in anaesthetized rat. This hypertensive effect is partially reversed (90%) by the prior administration of phentolamine (1 mg/kg) and abolished by pre-treatment with phentolamine (1 mg/kg) and atenolol (100 microg/kg). Neither tachiphylaxis nor any toxic effects were observed. These experimental findings suggest an interaction between C. pilosa and the peripheral adrenergic system (particularly with alpha1 and beta1 receptors); the structure of the bioactive glucosides could be important in evoking this effect. PMID- 11482758 TI - Trichilia roka Chiov. (Meliaceae): pharmacognostic researches. AB - The dried aqueous extract of Trichilia roka Chiov. (Meliaceae) root was evaluated for its potential antipyretic activity on yeast-induced hyperthermia in rats. The drug showed a significant reduction of body temperature when administered orally at the doses of 0.25, 0.5 and 1.0 g/kg. The antipyretic activity of T. roka was compared to indomethacin treatment (50 mg/kg), used as a reference drug. The results of this study confirm the validity of traditional usage of T. roka as an antipyretic agent. Moreover, micromorphological investigations were carried out by scanning electron microscopy obtaining useful phytognostic elements for the correct identification of the drugs both in scraped and powdered forms because this is of great interest for quality control in basic research and drug production, especially for imported items and for raw material sold by traditional herborists. PMID- 11482759 TI - Micromorphological investigations on Entada africana Guill. et Perr. (Mimosaceae). AB - Entada africana Guill. et Perr., known in Mali by the common local name 'Samanere' in the Bambara language, is one of the traditional Malian medicines prescribed for many illnesses. In the present investigation, micromorphological studies were carried out by scanning electron microscopy on the roots and the leaves. The correct identification of the morphological characters of drugs is of great interest for quality control in basic research and drug production, especially for imported items and for raw materials sold by traditional herbalists. PMID- 11482760 TI - Quality control of aromatic drugs reported in European Pharmacopoeia 3rd edition. AB - The apparent current explosion of interest and commercial activity in the area of herbal products should be followed by accurate quality control. In this work, the authors carried out the specific quality assays of all the aromatic drugs reported in European Pharmacopoeia. For each drug, several samples of different brands were bought in pharmacies, herbalist's shops, or supermarkets. The worst results were obtained in the assay for the essential oil content and this is very negative as the medicinal activities of these drugs are ascribed to their essential oil content. The samples of different brands yielded very different results from a qualitative point of view. In most cases, the analyzed samples were very far from the acceptable qualitative standards that they definitely lacked any health benefits. PMID- 11482761 TI - Effects of Calotropis procera on oestrous cycle and on oestrogenic functionality in rats. AB - Effects of ethanolic and aqueous extracts of Calotropis procera (Ait.) R.Br. roots, have been studied on oestrous cycle and on some parameters of oestrogenic functionality in rats. Both extracts have been shown to interrupt the normal oestrous cycle in 60 and 80%, respectively, of rats treated. The rats exhibited prolonged dioestrous stage of the oestrous cycle with consequent temporary inhibition of ovulation. The contemporary administration of commercial oestro progestinic preparation exhibited the same effects in 100% of rats treated. However, the extracts have not demonstrated to possess oestrogenic activity when tested in immature female bilaterally ovariectomized rats. PMID- 11482762 TI - Oxidative stress and usnic acid content in Parmelia caperata and Parmelia soredians (Lichenes). AB - High light levels together with Paraquat treatment or exposure to pollutants (e.g. SO2) can cause oxidative stress in epiphytic lichens. In some cases, a significant increase in ascorbic acid and other antioxidant metabolites, sometimes occurring in lichens only, was observed. In this study, usnic acid was measured by HPLC in Parmelia caperata and Parmelia soredians thalli treated with Paraquat, a herbicide which transfers electrons from various transport systems to oxygen, producing O2*- superoxide radicals. In light, Paraquat acts by generating active oxygen species within the chloroplast, thus simulating the oxidative component of environmental stress. The significant increase in the usnic acid content measured in P. soredians thalli (+ 36.3%) is in line with the hypothesis that it has an antioxidant action, but it is in contrast to the limited increase measured in P. caperata thalli (+ 13.7%). These apparently contradictory results confirm those found in the literature, which attribute different functions to usnic acid depending on the lichen species considered and on whether they have other detoxifying substances in their thalli. These studies are of potential application-oriented interest in relation to research into new active principles to be used in the pharmaceutical, food and cosmetic fields and/or in environmental biomonitoring. PMID- 11482763 TI - M1 receptor activation is a requirement for arecoline analgesia. AB - Arecoline. a drug obtained from the Areca Catechu L., induced a dose-dependent antinociception (0.3-1 mg kg(-1) i.p.) which was prevented by the muscarinic antagonists pirenzepine (0.1 microg per mouse i.c.v.) and S-(-)-ET-126 (0.01 microg per mouse i.c.v.). A dose-dependent inhibition of the antinociception induced by arecoline was observed after inactivation of the M1 gene by an antisense oligodeoxyribonucleotide (aODN). This effect was detected 24 h after the last i.c.v. injection of aODN. These results indicate that arecoline antinociception is mediated by the activation of central M1 muscarinic receptors. PMID- 11482764 TI - Local anaesthetic activity of beta-caryophyllene. AB - In this work we studied the local anaesthetic activity of beta-caryophyllene, one of the main components of clove oil obtained from the dried flower-buds of Syzygium aromaticum (Myrtaceae family). We compared its activity to a chemically related compound, caryophyllene oxide. Anaesthetic activity was evaluated in vivo in the rabbit conjunctival reflex test and in vitro in a rat phrenic nerve hemidiaphragm preparation. Beta-caryophyllene (10(-4) - 1 microg/ml), but not caryophyllene oxide, was able to reduce drastically, in a dose-dependent manner, the electrically evoked contractions of the rat phrenic hemidiaphragm. In the rabbit, conjunctival reflex test treatment with a solution of beta-caryophyllene (10-1000 microg/ml) allowed a dose-dependent increase in the number of stimuli necessary to provoke the reflex. As in the in vitro results, caryophyllene oxide was ineffective also in the in vivo test. In conclusion, these data evidence the local anaesthetic activity of beta-caryophyllene, which appears to be strictly dependent on its chemical structure. PMID- 11482765 TI - A decision support system for the management of accidental mushroom and plant poisoning. AB - In this paper the discussion focuses on a decision support system to be used as a tool in the treatment of poisoning cases attributable to mushrooms, plants and animals. In this first release, attention is focused on the risks related to fungi. Problems involved with mushroom poisoning and identification are analyzed to highlight which elements or characters must be taken into account in devising a computerized expert system. The components of such a system are presented, the different approaches are discussed and the choices made are motivated. Some preliminary results are also presented. PMID- 11482766 TI - The distribution of minerals and flavonoids in the tea plant (Camellia sinensis). AB - Camellia sinensis is a plant growing in India, Sri Lanka, Java, Japan and its properties were known 4000 years ago. Since then, traditional Chinese medicine has recommended this plant for headaches, body aches and pains, digestion, depression, detoxification, as an energiser and, in general, to prolong life. Tea contains volatile oils, vitamins, minerals, purines, polyphenols, particularly carechins. We have analysed ten commercial teas from various countries to determine their mineral composition and we have analysed a green tea, an Oolong tea and a White tea to determine their polyphenols and flavonoids content. Our study shows that the variation of mineral composition, polyphenols and flavonoids are linked to different origins of the plant. For the determination of phenols compounds and flavonoids we used an HPLC apparatus and for mineral analysis an atomic absorption apparatus. PMID- 11482767 TI - Cytotoxicity evaluation of natural coptisine and synthesis of coptisine from berberine. AB - The crude extract (80% MeOH in water) of Chelidonii herba exhibited very interesting cytotoxicity against brine shrimp (Artemia salina Leach) nauplii and cultured human tumour cell in vitro, the colon carcinoma HT 29 (144 h treatment). Fractionation of the crude extract and bioassay-guided procedures showed that the cytotoxic and the antitumour activities were concentrated in the basic extract. On the basis of IR, MS and 1H NMR the compound responsible of the cytotoxic activity was determined to be coptisine. Cytotoxicity evaluation of coptisine was next extended to a panel of human and murine cell lines in comparison with the established antitumour drugs mitoxantrone, doxorubicin (Dx) and cisplatin (CDDP). Coptisine was cytotoxic on LoVo and HT 29 and less potent on L-1210, and it was partially crossresistant on the human tumour colon cell line resistant to Dx, LoVo/Dx, whereas it was not significantly crossresistant on the murine leukaemia cell line resistant to CDDP, L-1210/CDDP. Coptisine alkaloid was then synthesised in gram amount from commercial berberine. A four-step synthetic route was elaborated. The overall yield was about 8-10%. The structural identity of synthetic coptisine was verified by IR and NMR methods. A comparison of the cytotoxic effects on the human tumour colon cell line LoVo and on the murine leukaemia L1210 showed, for both natural and synthetic coptisines, a comparable cytotoxic activity more evident against HT 29 cell line and LoVo cell line, while the activity was lower against the L1210 cell line. PMID- 11482768 TI - Nepeta sibthorpii Bentham (Lamiaceae): micromorphological analysis of leaves and flowers. AB - Nepeta sibthorpii Bentham (Lamiaceae) is an herbaceous aromatic plant, growing in dry, usually rocky areas of Southern Greece. In this paper, we report the study of micromorphological features of the flowers and leaves of Nepeta sibthorpii Bentham. The analysis by means of scanning electron microscopy has permitted to characterize both the covering trichomatous garment, abundantly present on the calyx and on the leaves, and the glandular hairs, distributed on the adaxial leaf surface and on the outer surfaces of calyx and corolla, showing also the peculiar features of the epidermal cells, all useful diagnostic elements for the identification of this medicinal plant. PMID- 11482769 TI - Isolation of praeruptorins A and B from Peucedanum praeruptorum Dunn. and their general pharmacological evaluation in comparison with extracts of the drug. AB - The root of Peucedanum praeruptorum Dunn. was extracted with solvents at different polarity obtaining three chemical fractions: aqueous (H2O), n-butanol (BuOH) and ethyl acetate (AcOEt). From AcOEt praeruptorins A and B were isolated by column chromatography on silica gel, using toluene/ethyl acetate as eluent, and identified by 1H and 13C NMR analysis. The extracts and the praeruptorins were tested for gross behavioural effects and acute toxicity in mice; the cytotoxicity on Artemia salina Leach and the antimicrobial activity were also evaluated. None of the tested substances evoked behavioural effects or acute toxicity after oral administration in mice; delayed mortality was observed with AcOEt and praeruptorin A only after intraperitoneal administration of high doses (1 g/kg). In Artemia salina test AcOEt, and praeruptorins A and B had LC50 values of 40.2, 121.2 and 34.5 microg/ml, respectively. AcOEt and praeruptorin A showed antimicrobial activity on Streptococcus agalactiae; their MIC values were 250 and 100 microg/ml, respectively. PMID- 11482770 TI - Microporous material from kanemite for drug inclusion and release. AB - A microporous material obtained from kanemite, a layered polysilicate, was studied in order to investigate its feasibility of including drugs and then releasing them. Diphenydramine hydrochloride was chosen as a model drug. The preparation of the microporous material and its loading with the drug are described. As kanemite is able to intercalate anions between its layers, the intercalation compound of diphenydramine and kanemite was also prepared. Both the drug-loaded microporous material and the intercalation compound were submitted to dissolution tests at pH 7.5. The drug release profiles from these two different materials and from a physical mixture were compared. PMID- 11482771 TI - Stability studies of new cosmetic formulations with vegetable extracts as functional agents. AB - A formulation study, using increasing amounts of Sepigel 305 as an emulsifier, has been carried out to find new O/W emulsions and value their stability also in presence of vegetable extracts. Stability results have been compared with those obtained from formulations described in the National Formulary of Italian Pharmacopoeia X and functionalised by us with the same vegetable extracts. By using centrifugation and accelerated ageing tests capable of bringing out the gelling and thermostability properties of Sepigel 305, the study emphasised that the new gel emulsions have a greater stability compared to the other formulations. PMID- 11482772 TI - Chemical composition and cytotoxic and antimicrobial activity of Calycotome villosa (Poiret) link leaves. AB - The chemical composition of the essential oil and methanol extract of Calycotome villosa (Poiret) Link leaves collected in Sardinia (Italy) has been studied by analytical and spectroscopic methods. Falcarinol and some alcohols, terpenes, furan derivatives, and paraffins have been isolated from the essential oil. Thirteen alkaloids and falcarinol have been identified in the chloroform fraction of the basic methanol extract. Six flavonoids and four anthraquinones have been isolated in the chloroform fraction after acidification of the basic methanol extract. The cytotoxic and antimicrobial activities have also been evaluated. The essential oil, the methanol extract in toto, and the fraction of the basic extract showed strong cytotoxicity, whereas the fraction of the acid extract showed lower cytotoxicity. Furthermore, this fraction showed good antibacterial activity against Staphylococcus aureus, Bacillus lentus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Providencia rettgeri, and Morganella morganii. It can therefore be stated that this plant's cytotoxicity is prevalently due to falcarinol. PMID- 11482773 TI - Rheological characterization of hydroalcoholic gels--15% ethanol--of Carbopol Ultrez 10. AB - The rheological behaviour of Carbopol Ultrez 10 ethanolic (15%) gels was studied as a function of two main variables: pH (4.0-7.0) and the polymer concentration (0.1-0.5%). Flow curves of all hydroalcoholic gels were adjusted to the Ostwald model. Sigmoidal dose response functions were obtained to describe the pH dependence of consistency index of the systems. The concentration dependence of flow indexes was best adjusted to one-phase exponential decay functions. Finally, by substituting in the power law expression, an empirical model is given to estimate viscosities of gels for any value of pH and polymer concentration in the ranges evaluated. As a result, the influence of the variables on polymer network is meaningfully assessed by means of the obtained empirical parameters: bottom value (Ymax), pH value required for a 50% polymer network development (pH50), and asymptotic flow index value for the fully structured gels (n(min)). PMID- 11482774 TI - Viscosity and temperature relationship in ethanol/water mixtures gelified with Carbopol Ultrez 10. AB - The rheological characterization of Carbopol Ultrez 10 ethanolic (15%) gels is complemented by studying flow behavior as a function of temperature (20-50 degrees C) at different pH (4.0-7.0) and polymer concentrations (0.1-0.5%). Flow curves were adjusted to the Ostwald model, showing a fall in viscosity, together with an enhancement in flow index with increasing temperature. Calculated flow activation energies at constant shear rate (Egamma) were found to be related to the final pH of hydroalcoholic gels. Nevertheless, at the polymer concentration range evaluated, no significant correlations were obtained between activation energy and concentration. In general, the Carbopol Ultrez 10 gels exhibit little viscosity change under the temperature variations of normal use and storage, which minimizes possible unacceptable changes in the product's characteristics. PMID- 11482775 TI - Effect of the exposure to gentamicin and diltiazem on the permeability of model membranes. AB - Preliminary observations showed that the calcium-antagonist diltiazem enhances the 'in vitro' bactericidal action of the aminoglycoside gentamicin, especially against Gram-positive bacteria. To verify if a non-specific interaction of these two drugs with biomembranes may play a role in their synergic effect on bacterial cells, we have studied the effect of exposure to gentamicin, in the absence or presence of diltiazem, on the release of carboxyfluorescein (CF) trapped in phosphatidylcholine (PC) unilamellar vesicles (LUVs) used as model membranes. A significant leakage of trapped CF from PC LUVs was registered when liposomes were treated with gentamicin and diltiazem together, employed at doses (50 and 100 microg/ml, respectively) unable to affect CF release if applied alone; the combined effect of gentamicin and diltiazem was synergic and not cumulative. The present findings demonstrate that the simultaneous exposure to gentamicin and diltiazem may induce significant alterations in the permeability of phospholipid membranes and, so, very likely, in functional properties of bacterial membranes, targets of their action. PMID- 11482776 TI - Efficacy of Hypericum and Calendula oils in the epithelial reconstruction of surgical wounds in childbirth with caesarean section. AB - Following studies on the properties of spontaneous plants in Sardinia we have evaluated the tissue regenerating action of a mixture of oily extracts of Hypericum perforatum and Calendula arvensis on surgical wounds from childbirth with caesarean section. PMID- 11482777 TI - Anti-inflammatory and analgesic activity of Hypericum empetrifolium Willd. (Guttiferae). AB - A methanolic extract of Hypericum empetrifolium Willd. was evaluated for anti inflammatory properties in rats (subplantar edema induced by carrageenan) and analgesic effects in mice (hot plate and writhing tests). Our results showed that the methanolic extract exhibits a significant anti-inflammatory activity and analgesic effects only in one of the experimental models (writhing test). Therefore, we may suppose that the methanolic extract of H. empetrifolium is active against inflammatory pain. PMID- 11482778 TI - Anticonvulsant and sedative effects of some 5-substituted bromopyrazolinic spirobarbiturates. AB - We studied in mice the neuropharmacological effect of some 5-substituted bromopyrazolinic spirobarbiturates. LD50 and CL50 values were estimated for all the investigated compounds. The effect on potentiation of sodium pentobarbital activity and on generalised tonic-clonic seizures produced by PTZ on mice was studied. All the compounds had the same pharmacological profile, they extended sleeping-time induced by sodium pentobarbital and showed protection against PTZ induced convulsions, decreasing the death rate. PMID- 11482779 TI - Oxidants and antioxidants in long-term haemodialysis patients. AB - Survival for decades is now possible in end-stage renal disease patients (ESRD) treated with haemodialysis (HD). Long-term survivors may present dialysis-related pathology (DRP). Alterations in lipid metabolism and oxidative stress are recognized as important risk factors that could be prevented or reduced by optimal therapy. We have studied markers of oxidative stress in patients receiving HD treatment for more than 20 years. In order to evaluate a preventive intervention against oxidative damage we measured the factors implied for the prooxidative and antioxidative mechanisms in haemodialysis patients. Ten long term HD survivors (HD duration: 274.2 months) and ten patients with recent onset of HD (HD duration: 17.8 months), had blood drawn for plasma vitamins A and E, malondialdehyde (MDA), plasma and RBC glutathione peroxidase (GPx), RBC superoxide dismutase (SOD), plasma and erythrocyte glutathione reductase (GSSG R), oxidized and reduced glutathione (GSH) assessment. Despite normal levels of antioxidant vitamins, an usual finding in this setting, increased MDA, and oxidized GSH, and decreased plasma GPx and reduced GSH show that oxidant stress is markedly present in both recent onset and long-term HD patients. It would appear highly advantageous to reduce complications of long-term dialysis patients with preventing modalities. PMID- 11482780 TI - Basic fibroblast growth factor and keratinocyte growth factor over-expression in benign prostatic hyperplasia. AB - The prostate growth is under the indirect control of androgens through the medium of many growth factors. The fibroblast growth factors (FGFs) seem to play an important part in stimulating the development of this organ. In this work, the expression of two FGFs: bFGF (or FGF2) and KGF (or FGF7), was studied in RT-PCR and semi-quantified in densitometry. Both genes expression was increased in BPH in comparison with normal prostates. A two to three times and a 1.5 to four times over-expressions were observed for bFGF and KGF, respectively. An over-expression of these growth factors could lead to a cell multiplication resulting in a pathological development of the prostate size. Moreover, bFGF and KGF act in parallel in the prostate, one stimulating the prostatic stroma and the other one stimulating the epithelium. These two growth factors could participate in the increase of the two tissues constituting the prostate. PMID- 11482781 TI - Low-density lipoprotein (LDL) behavior after in vitro oxidation in three groups of diabetics. AB - Diabetes is associated with increased morbidity and mortality resulting from cardiovascular disease. It has been established that oxidized LDLs are involved in the genesis of atherosclerosis. We have studied LDL oxidizability in three types of diabetics: insulin-dependent diabetes mellitus (IDDM), non-insulin dependent diabetes mellitus (NIDDM) and insulin-treated diabetes mellitus type 2 (ITDM2) and a control group. LDLs have been isolated using ultracentrifugation and oxidized by addition of cupric chloride. With the oxidation kinetic, we calculated the lag time and the oxidation rate. Total fatty acids, alpha tocopherol, and malondialdehyde (MDA) have been measured in native and oxidized LDLs. Diabetics have a significantly lower lag time and a lower level of alpha tocopherol. Oxidized LDLs of diabetics show an important decrease of their polyunsaturated fatty acids with an increase of MDA compared to the control. Our study demonstrates a higher susceptibility to oxidation of LDL from diabetics; this can be explained by alteration in LDL composition or by the oxidative process occurring in this disease. PMID- 11482782 TI - Pharmaco-toxicological screening of commercially available Italian natural mineral waters. AB - The consumption of natural mineral water has increased enormously during the past few years, yet doubts are arising about the real utility of using such water instead of ordinary drinking water (tap water). Mineral water's 'special' mineral composition might have properties favourable to health, which should be assessed by clinical and pharmacological analyses. A comparative pharmaco-toxicological study was performed on 14 commercially available Italian natural mineral waters with a wide range of mineral content. The waters were microbiologically analysed and the Allium cepa test done; in addition, Irwin, charcoal meal and diuresis tests were also performed on laboratory animals. No toxicological signs were recorded at the Allium cepa assay for any but the water with the highest mineral content. The diuretic effect was comparable to that of control tap water. Waters with high magnesium content significantly enhanced intestinal motility while at the Irwin test none of the water samples evoked behavioural changes. All the tested samples were microbiologically pure. In conclusion, mineral water can be an alternative to aqueduct water in places where the latter comes from superficial water and has to be subjected to hard potabilisation techniques; water from the 'Acqua Marcia' aqueduct (control) was found to have characteristics comparable to the mineral waters studied. PMID- 11482783 TI - Anti-proliferative effect on a prostatic epithelial cell line (PZ-HPV-7) by Epilobium angustifolium L. AB - Symptomatic benign prostatic hyperplasia (BPH) is a common condition in elderly men and has a significant impact on their daily lives. The drugs prescribed for treatment include alpha1-blockers, 5-alpha-reductase inhibitors and plant preparations. Epilobium angustifolium L. is deemed to be helpful in BPH therapy, although there is less information regarding the mechanism of its biological activity. The present study evaluated the effect of E. angustifolium extract on human prostatic epithelial cells (PZ-HPV-7). The exposure to E. angustifolium extract induced a marked inhibition of cell growth in all tested conditions. The anti-proliferative effect observed in in vitro systems clearly indicates a biologically relevant effect of compounds present in the extract. Considering these results, the use in traditional medicine of E. angustifolium extract against BPH seems to be justified. However, further experimental studies are needed to determine the biochemical mechanism of the action and the clinical value of the E. angustifolium extract. PMID- 11482784 TI - Comparative evaluation of St. John's wort from different Italian regions. AB - Flowers and fruits of St. John's wort collected in different Italian regions were evaluated for their naphtodianthrone (hypericin and pseudohypericin), phloroglucinol (hyperforin and adhyperforin) and flavonol (rutin, quercetin, quercitrin, isoquercitrin and hyperoside) content. The quantitative evaluation was performed by HPLC-DAD. The crude drug collected at the fruit ripening period had the highest content in phloroglucinols and the lowest level of both naphtodianthrones and flavonols. Phloroglucinols peaked in the samples collected in Puglia followed by Lombardia and Veneto, while hypericins and flavonols were highest in the samples harvested in Sardegna and Trentino. PMID- 11482785 TI - Antioxidant activity of different fractions of Spirulina platensis protean extract. AB - Spirulina platensis, planktonic blue-green algae, is gaining increasing attention because of its nutritional and medicinal properties. This microalgae contains phycobiliproteins (phycocyanin and allophycocyanin). Previous reports from our laboratory have shown that a protean extract of S. platensis is a potent free radical scavenger (hydroxyl and peroxyl radicals) and inhibits microsomal lipid peroxidation. The aim of this study was to purify and characterize phycocyanin of S. platensis. Besides, we tried to demonstrate that one of the main components responsible for this antioxidant activity is a biliprotein phycocyanin. For this purpose, we studied the antioxidant activity of different fractions obtained during the phycocyanin purification process, through the scavenger activity of hydroxyl radical. We also observed that an increase in phycocyanin content was related to an increase in the antioxidant activity in different fractions, and therefore phycobiliprotein phycocyanin is the component mainly responsible for the antioxidant activity. PMID- 11482786 TI - Antiulcerogenic activity of Lippia alba (Mill.) N. E. Brown (Verbenaceae). AB - Lippia alba (Mill.) N. E. Brown Verbenaceae, known popularly as 'Juanilama' or 'Salvia Sija', is prized widely in folk medicine in Guatemala. Its leaves are employed as an infusion and decoction as a remedy for stomach problems, dysentery, colds and cough, febrifuge, as well as a sedative and in spasmolitic remedies. The present study reports the effects of the infusion of L. alba on the rat gastric mucosa. The following behavioural parameters were evaluated: (a) gastric irritancy test in Wistar rats; (b) antiulcer activity, short term and long term; (c) acid secretion; (d) measurement of total proteins; (e) estimation of total protein bound and nonprotein sulfhydryl groups. Ranitidine (100 mg/kg, p.o.) was used as the reference antiulcer drug. Oral treatment with the infusion (12.5 g dry plant/kg) did not cause gastric irritancy in the rats treated during five consecutive days. In addition, the oral administration of L. alba was found to be effective at preventing gastric ulceration induced by indomethacin (50 mg/kg, p.o.) in rats in the short term (1 day) and long term (5 days). PMID- 11482787 TI - Comparison of planar chromatographic methods (TLC, OPLC, AMD) applied to essential oils of wild thyme and seven chemotypes of thyme. AB - Essential oils analysis is more often realized by gas chromatography. However, thin-layer chromatography (TLC) remains the reference for Pharmacopoeia. Nevertheless classical TLC has its own limitations but it is always a good technique because it is simple, rapid and less expensive. Actually the reproducibility, the separation quality and the possibility to obtain good and reproducible quantitative determinations have been improved significantly with automated sample applicator, scanner densitometers and two new chromatographic planar methods: the optimum performance laminar chromatography (OPLC) and automated multiple development (AMD). In this work, we show and compare the performance of these methods and TLC through a study of seven thyme chemotypes and wild thyme essential oil. PMID- 11482788 TI - Development of a cream from a self-emulsifying base and moisturizing actives. AB - This research study is based on the design and development of a semisolid emulsion system whose novel self-emulsifying base and preferentially moisturizing actives were investigated to find out whether the system mentioned could be used as a dermatological treatment for highly sensitive skins, including atopic ones. Thus, one of the main objectives of the present study consisted of in vivo evaluation of its effectiveness by means of non-invasive assessment techniques currently employed in cosmetology. Due to the fact that the new formula is, in principle, designed for skins that could present any kind of alteration, the current study was focused on rheological parameters of viscosity, thixotropy, and extensibility to guarantee not only an accurate assessment of composition but also a comfortable and safe application on skin. PMID- 11482789 TI - Preliminary screening of five ethnomedicinal plants of Guatemala. AB - We performed the Irwin test on some different extracts of the aerial parts of Tridax procumbens L., of the leaves of Neurolaena lobata (L.) R. Br., of the bark and leaves of Byrsonima crassifolia (L.) Kunth. and Gliricidia sepium Jacq. Walp. and of the root and leaves of Petiveria alliacea L. At a dosage of 1.25 g extract/100 g dried plant, the aqueous extracts of bark and leaves of Byrsonima crassifolia (L.) Kunth. and G. sepium Jacq. Walp. showed higher activity: decrease in motor activity, back tonus, reversible parpebral ptosis. catalepsy and strong hypothermia. PMID- 11482790 TI - Mucopolysaccharides in aqueous solutions: effect of ionic strength on titration curves. AB - We study the changes taking place in hyaluronic acid, chondroitin 4-sulfate (C4 S) and condroitin 6-sulfate (C6-S), at ionic strengths of 0.10, 0.15, and 0.20 in NaCl, in a neutralization process in aqueous solution. We apply the equation of Henderson Hasselbalch modified for polyelectrolytes and evaluate the changes in the electrostatic free energy starting from the pK curves as a function of the dissociation degree. For a dissociation degree next to 0.4 corresponding to the COOH group of the hyaluronic acid, we observed a change in the conformation of the three glycosaminoglycans studied. This conformational change takes place as a consequence of the break of intramolecular links and the beginning of the ionization process. The macromolecules in solution show a structure of random coil sufficiently expanded so that the interaction among the close ionizable groups is negligible. PMID- 11482791 TI - Changes in the flow properties of phospholipid dispersions induced by procaine hydrochloride. Effect of pH and temperature. AB - Since local anaesthetics are known to interact with membrane lipids, we have examined the changes taken place by procaine hydrochloride in lipid matrices as a function of pH. Rheological methods might give useful information on the association of this anaesthetic with soybean lecithin. The procaine interacted with negatively charged phospholipid polar head groups at pH 4. This system exhibits a loosening in the tight arrangement of phospholipid molecules caused by the addition of procaine as a function of this anaesthetic's concentration. The flow enthalpy values as a function of procaine-lipid ratio shows biphasic behaviour and suggests a phase transition when the anaesthetic concentration goes from 10 to 14 mM and temperatures dip below 10 degrees C. PMID- 11482792 TI - Changing health inequalities in the Nordic countries? AB - The Nordic countries, referring here to Denmark, Finland, Norway, and Sweden, have often been viewed as a group of countries with many features in common, such as geographical location, history, culture, religion, language, and economic and political structures. It has also been habitual to refer to a "Nordic model" of welfare states comprising a large public sector, active labour market policies, high costs for social welfare as well as high taxes, and a general commitment to social equality. Recent research suggests that much of this "Nordicness" appears to remain despite the fact that the Nordic countries have experienced quite different changes during the 1980s and 1990s. How this relates to changes in health inequalities is in the focus of this supplement. PMID- 11482793 TI - Social differences in illness and health-related exclusion from the labour market in Denmark from 1987 to 1994. AB - OBJECTIVE: The objectives were first to analyse differences in long-standing illness, limiting long-standing illness, and perceived health as below good in relation to different indicators of social class over time; and secondly to analyse the association of long-standing illness, educational level, age, and gender with employment status. SUBJECTS AND METHODS: Self-reported information on illness, educational level, employment status, and occupational class was obtained in two nationally representative Danish health interview surveys in 1987 (n=4753) and 1994 (n=4668). RESULTS: There was a marked increase in long-standing illness from 1987 (33%) to 1994 (38%), especially among women with higher education (13% in 1987 to 26% in 1994). The prevalence of perceived health below good was unchanged (c. 20%) during the same period. Among employed men in 1987 the lowest prevalence of long-standing illness was seen in upper salaried employees (21%) and the highest in skilled workers (35%). In 1994, the difference had diminished and the prevalence rates for the same classes were 28% and 29%, respectively. Among employed women, a similar pattern was seen. The lowest rate in 1987 was seen among self-employed with subordinates (15%) and the highest in self-employed without subordinates (28%). In 1994, the difference was reduced with prevalence rates of 23% and 32%, respectively. In addition, substantial differences in health status between groups with different educational backgrounds were found. The proportion of the population with long-standing illnesses was clearly higher in the group with low education compared with the group with high education. A similar social gradient was found for perceived health as below good. Those with the highest odds ratio of being a disability pensioner are women with long-standing illness, only basic education, and above 55 years of age. CONCLUSION: In conclusion, the study showed that the prevalence of long-standing illness increased from the mid-1980s to the mid-1990s, whereas perceived health below good was unchanged. In 1994, there was a remarkable difference in health between employed and non-employed people, indicating a health-related exclusion from the labour market. This may explain why, in 1994, smaller occupational class differences were found in the prevalence of long standing illness among employed people compared with the findings in 1987, whereas the health differences remained in the different educational groups. Women above 54 years of age with basic education only and long-standing illness have the highest odds ratio of permanent exclusion from the labour market. PMID- 11482794 TI - Changing health inequalities in a changing society? Sweden in the mid-1980s and mid-1990s. AB - Whereas the end of the 1980s was characterized by an economic boom, the early 1990s saw the worst recession since the 1930s. In Sweden, the crisis that started in the fall of 1991 and culminated in 1995 meant dramatically increased unemployment rates followed by cutbacks in welfare state programs. In addition, other major changes in economic and political conditions have taken place during this period, including tax reforms and EU membership. Although public health as well as health inequalities are likely to be linked with these kinds of macro changes, it is unclear what types of changes in health and health inequalities one would expect. In this paper analyses of Swedish data on health inequalities in the periods 1986-87 and 1994-95 are undertaken on the basis of the Swedish Surveys of Living Conditions. The main finding is that overall health levels as well as differences in health between men and women, different age groups, educational groups, social classes, and employment status groups have remained constant. PMID- 11482795 TI - Structural changes and social inequalities in health in Finland, 1986-1994. AB - BACKGROUND: This paper seeks to examine changes in health inequalities in Finland from the mid-1980s to the mid-1990s. To improve our understanding of the contribution of employment status changes to class and educational differences in health within the context of changing labour market conditions, we examined the differences in ill health among social classes, educational groups, and employment status groups. METHODS: The data was derived from nation-wide Finnish Surveys on Living Conditions from 1986 and 1994. Analyses included the age range 25-64 years. Health was measured using limiting long-standing illness and perceived health as below good. Results are presented as age-standardized prevalence percentages and odds ratios from logistic regression analyses. RESULTS: The pattern and size of relative social inequalities in ill health have remained generally stable during the eight-year study period in Finland. Differences between social classes have changed only slightly. Differences between educational groups have declined somewhat among men, but have remained stable among women. Compared with the employed, health among the unemployed, housewives, and pensioners has improved for both men and women. Analysing social class differences within both the employed and the non-employed showed only negligible changes. CONCLUSIONS: Changes in social inequalities in health as indicated by class and educational differentials among Finnish men and women have mostly been negligible. The observed changes are likely to have been affected by the 1990s' labour market crisis in Finland. The rapidly increasing mass unemployment is unlikely to have been very individually selective in the short run. However, in the longer run, to the extent that unemployment remains high, this trend can be expected to change as re-entry to paid employment is likely to be more individually selective. PMID- 11482796 TI - Recent changes in social structure and health inequalities in Norway. AB - AIM: The article explores whether changes in social structure from the mid-1980s to the mid-1990s have been accompanied by any changes in ill health, both in the general population and in the association between socioeconomic positions and ill health. DATA AND METHODS: Data were derived from four interview surveys conducted by Statistics Norway: The Health Surveys from 1985 and 1995 and the Surveys of Level of Living from 1987 and 1995. These surveys provide nationwide representative samples of the adult population. The analyses are confined to the population between 25 and 64 years of age. Simple cross-tabulations, age standardized cross-tabulations, and logistic regression analysis were used. In addition, a more genuine measure of inequality was calculated--the so-called concentration index. RESULTS: In the examined period, the educational level of the population gradually improved, and more people had white collar jobs. On the other hand, the decade was a turbulent one in economic terms, and more people experienced unemployment and early retirement from work during the early 1990s than during the preceding period. Nevertheless, levels of ill health, as measured in terms of self-reported limiting long-standing illness and global self perceived health, were only marginally altered. Also, health differentials according to socioeconomic positions were only trivially changed during this period. CONCLUSION: In the mid-1990s, as in the mid-1980s, lower socioeconomic positions, such as unemployment and early retirement, only basic education, and workers' occupations, are generally connected to poorer health. No substantial change in this association is detected in this period. PMID- 11482797 TI - Effects of repeated exposure of rats to JP-5 or JP-8 jet fuel vapor on neurobehavioral capacity and neurotransmitter levels. AB - The U.S. Naval Service is anticipating transition from the nearly exclusive use of JP-5 jet fuel to predominant use of JP-8, consistent with the primary utilization by the U.S. Army, U.S. Air Force, and the militaries of most NATO countries. To compare the relative risk of repeated exposure to JP-5 versus JP-8 vapor, groups of 32 male Sprague-Dawley rats each were exposed for 6 h/d, 5 d/wk for 6 wk (180 h) to JP-8 jet fuel vapor (1,000 +/- 10% mg/m3), IP-5 vapor (1,200 +/- 10% mg/m3), or room air control conditions. Following a 65-d rest period, rats completed 10 tests selected from the Neurobehavioral Toxicity Assessment Battery (NTAB) to evaluate changes in performance capacity. Repeated exposure to JP-5 resulted in significant effects on only one test, forelimb grip strength (FGS), while exposure to JP-8 vapor resulted in a significant difference versus controls on appetitive reinforcer approach sensitization (ARAS). Rats were further evaluated for concentrations of major neurotransmitters and metabolites in five brain regions and in the blood serum. Levels of dopamine, the dopamine metabolite dihydroxyphenylacetic acid (DOPAC), and the serotonin metabolite homovanillic acid (HVA) were significantly modulated in various brain regions, as measured 85+ d postexposure. Similarly, serum levels of the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) were differentially modulated following JP-8 or JP-5 exposure. Results are compared to previously published research evaluating the neurotoxicity of repeated exposure to other hydrocarbon fuels and solvents. PMID- 11482798 TI - Effects of diesel exhaust particles on blood pressure in rats. AB - The effects of diesel exhaust particles (DEP) on pulmonary functions and consequent diseases are well known, but there have been few reports concerning involvement of the cardiovascular system. In order to assess a direct action of DEP on cardiac tissue, the effects on blood pressure of intravenous administration of 12 or 120 mg/kg DEP to anesthetized rats were studied for a 15 min period. DEP (120 mg/kg) significantly lowered blood pressure for 25 s with no signs of arrhythmia or mortality, a phenomenon seen in guinea pigs. After 25 s blood pressure gradually returned to control levels and was maintained for 15 min. The 12-mg/kg DEP concentration did not markedly affect rat blood pressure. Pretreatment with atropine (24 mg/kg) blocked the DEP-induced fall in blood pressure, while pretreatment with propranolol (48 mg/kg) proved ineffective against DEP, suggesting involvement of the parasympathetic system. Data show that the rat is less sensitive to DEP-induced effects on blood pressure and may be a poor model to reflect cardiovascular changes. PMID- 11482799 TI - Characterization of volatile organic compounds in smoke at municipal structural fires. AB - The objective of this study was to characterize volatile organic compounds (VOCs) found at municipal structural fires in order to identify sources of long-term health risks to firefighters, which may be contributing factors in heart disease and cancer. Firefighters collected air into evacuated Summa canisters inside burning buildings at nine municipal structural fires under conditions where they judged that at least some firefighters might remove their self-contained breathing apparatus masks. Volatile organic compounds were identified and quantified for 144 target compounds using cryogenic preconcentration and gas chromatography/mass spectral detection (GC/MSD) methodology operating in selected ion monitoring mode. Samples were also analyzed in SCAN mode and examined for the appearance of substances that were not present in the instrument standard calibration mixture. The spectra of municipal structural fires were surprisingly similar and remarkable for their simplicity, which was largely due to the dominating presence of benzene along with toluene and naphthalene. Propene and 1,3-butadiene were found in all of the fires, and styrene and other alkyl substituted benzene compounds were frequently identified. Similar "fingerprints" of the same 14 substances (propene, benzene, xylenes, 1-butene/2-methylpropene, toluene, propane, 1,2-butadiene, 2-methylbutane, ethylbenzene, naphthalene, styrene, cyclopentene, 1-methylcyclopentene, isopropylbenzene) previously identified at experimental fires burning various solid combustible materials were also found at municipal structural fires, accounting for 76.8% of the total VOCs measured. Statistically significant positive correlations were found between increasing levels of benzene and levels of propene, the xylenes, toluene, 1 butene/2-methylpropene, 1,3-butadiene, and naphthalene. Given the toxicity/carcinogenicity of those VOCs that were found in the highest concentrations, particularly benzene, 1,3-butadiene, and styrene, further investigation of VOC exposures of firefighters is warranted. Benzene, or its metabolic product s-phenylmercapturic acid in urine, was identified as a suitable chemical marker for firefighter exposure to combustion products. PMID- 11482800 TI - Development of porous clay-based composites for the sorption of lead from water. AB - Lead contamination of water is a major health hazard, as illustrated by the fact that exposure to this metal has been associated with death and disease in humans, birds, and animals. The present research was aimed at the development of a porous, solid-phase sorbent that can be used in the remediation of lead contaminated water. A suitable sorbent was identified by screening various clays and other materials for their ability to effectively bind lead. The clay was adhered to a solid support using an aqueous solution of carboxymethyl cellulose. The binary composite was then tested for its ability to bind lead from solution, while providing void volume, increased surface area, and considerably enhanced hydraulic conductivity. The results suggested that a combination of sodium montmorillonite clay and carbon exhibited enhanced sorption of lead compared to carbon alone, and also supported the potential application of various combinations of sorbent materials. This value-added combination of clay, solid support, and adhesive will allow for the construction of column filtration systems that are multifunctional and capable of purifying large volumes of contaminated water. PMID- 11482801 TI - Primary ectopic meningioma. AB - Primary ectopic meningiomas are rare. A case of a 16-year-old male who presented with a large mass in temporofrontal region is reported. X-ray skull showed soft tissue shadow with hyperostosis of frontal and temporal bone. CT scan of brain demonstrated a markedly enhancing lesion and bony hyperostosis with no intracranial component. Total excision of tumour with hyperostotic bone was done. Patient is well without any evidence of recurrence two years after surgery. Relevant literature is reviewed. PMID- 11482802 TI - Ataxica telengiectasia (Louis-Bar syndrome). AB - A case of ataxia telengiectasia in a 7-year-old girl with symptoms for last 3 years has been reported in detail. The necessary literature has been reviewed with proper references. PMID- 11482803 TI - Glioblastoma multiformis, rapidity of growth--importance of repeat CT scan. AB - A case of glioblastoma multiformis (GBM) where lesion appeared within a month of normal CT scan is reported. It emphasises the importance of early CT scan after neurological deterioration particularly in older patients because of rapid growth of glioblastoma. PMID- 11482804 TI - Efficacy and safety of Ascoril in the management of cough--National Study Group report. AB - A total of 768 patients were recruited by 81 physicians and paediatricians all over the country in this National Study Group of 'Ascoril + Expectorant'. The results of this first large scale study of a cough formula indicates that 'Ascoril + Expectorant' is effective in controlling cough, breathlessness and decreasing the volume of sputum. No serious adverse events were noted. Ascoril was well accepted by the patients and its efficacy was rated very high by the physician. The National Study Group concludes that 'Ascoril + Expectorant' is highly effective in the management of cough associated with lower respiratory tract infection and COPDs. PMID- 11482805 TI - Epileptic and non-epileptic seizures. PMID- 11482806 TI - Headache--an Indian perspective. PMID- 11482807 TI - How important is history of unconsciousness in head injury patients? AB - A total of 100 patients of head injury were studied. They all underwent thorough clinical and neurological examination, skull radiography and CT scan of head. And with the help of all these parameters an evaluation was done to find out the importance of history of unconsciousness on risk of intracranial complications. A lot of controversies exist about detection of factors that make a patient of head injury a high risk candidate for developing intracranial complications. Young active population was the most commonly affected group in head injury with male preponderance at all ages. Roadside accident found to be the main cause of head trauma in adults while fall from height in paediatric age group. Although Glassgow Coma Index (GCI) was found to be a good predictor for intracranial complications as well as final outcome in patients with total score of 12 or less but it was not equally good in patients of minor head injury (GCI = 13-15). Similarly history of unconsciousness was not found to be correlating well with risk of intracranial complications but longer duration of unconsciousness was found to be a poor prognostic index. PMID- 11482808 TI - Studies on central nervous system function in diabetes mellitus. AB - Fifty-seven insulin dependent (IDDM) and non-insulin dependent (NIDDM) diabetic patients and 25 controls were studied. Patients with history of strokes, hypoglycaemia, hearing impairment, diabetic retinopathy, etc, were excluded. Clinical examination of central nervous system (CNS) and computerised tomography scan of brain were absolutely normal in all cases. Neuroelectrophysiological tests done were the visual evoked potential (VEP), brainstem auditory evoked response (BAER) and somatosensory evoked potential (SEP). The mean VEP latency was significantly raised in both NIDDM and IDDM compared with controls. The mean BAER and SEP latencies were both significantly raised in NIDDM but not in IDDM. The percentage of cases with abnormally raised CNS latencies were as follows: In NIDDM, VEP-16.7%, BAER-50% and SEP-26.7%; in IDDM, VEP-11.1%, BAER-14.8% and SEP 18.5%. Thus, subclinical CNS dysfunction is common in diabetes mellitus particularly in NIDDM and this can be reliably detected by measuring the CNS latencies, specially VEP. PMID- 11482809 TI - Stress management for patient and physician. AB - Starting with the definition of stress the author vividly describes physical response to stress, stress related illnesses, common symptoms and causes of stress and lastly stress management. His concluding remarks mention about some Reiki principles. PMID- 11482810 TI - Tea and health. AB - Tea, its health effects and other benefits have been described in detail with an introduction about the origin of tea. PMID- 11482811 TI - Neurocysticercal serodiagnosis--updated. AB - For diagnosis of neurocysticercosis the gold standard would be a stereotactic biopsy and histological confirmation which are not universally available or acceptable to patients, hence the necessity of immunodiagnosis. The authors have narrated beautifully the serodiagnostic aspects of neurocysticercosis in its updated form and have discussed the subject with proper references. PMID- 11482812 TI - Imaging transmitter release. I. Peeking at the steps preceding membrane fusion. AB - Over the recent year we have witnessed considerable advances in the study of neurotransmitter release. This progress has been severalfold as different techniques have allowed us to characterise many steps along the process of exocytosis, membrane fusion, formation of the fusion pore, and have given insight in the kinetics of release and membrane re-uptake. Patch clamping provided quantitative measurements of the capacitance changes as the membrane of the secretory vesicle is added to the surface of the cell during secretion, and the change in the opposite direction when membrane is retrieved back into the cell during exocytosis. Carbon-fibre microelectrodes have measured electrochemically the release of oxidisable transmitters into the extracellular space. Differential interference contrast microscopy has given us spatially resolved images of the cell surface during exocytosis; real-time images that are suggestive of bubbles breaking the surface of a boiling pot of water. The interest in novel techniques stems from the fact that existing approaches can provide only indirect evidence on the steps preceding membrane fusion. The vesicular dynamics just beneath the plasma membrane are out of the reach of capacitance measurements or amperometric detection. What we have needed is a tool that would allow us to look just below the cell surface. This much-needed tool appears to be evanescent-wave microscopy. This review describes how laser microscopy can be used to study exocytosis at single-vesicle resolution. A companion paper deals with the practical aspects of evanescent-wave imaging. PMID- 11482813 TI - Imaging transmitter release. II. A practical guide to evanescent-wave imaging. AB - Total internal reflection of a laser beam at an interface between two media of different refractive index sets up an evanescent wave field in the medium with lower refractive index. This near field decays over a distance of approximately lambda/5, lambda denoting the wavelength of light, and thus provides a convenient means for the confinement of fluorescence excitation to the near-interface region. Evanescent-wave excitation thereby permits, for example, the observation of individual fluorophores at the surface despite the presence of high concentrations in bulk solution. Although evanescent-wave excitation of fluorescence and the related technique of surface-plasmon resonance have a long record in the study of chemical reactions at surfaces, adsorption kinetics or spectroscopy, their potential for biomedical studies is only gradually emerging. Evanescent-wave microscopy provides high-contrast images of the near-membrane region of cells grown on a glass substrate at unprecedented resolution. At present, no commercial equipment is available for evanescent-wave microscopy. This review aims at readers who want to modify their fluorescence microscope to include an evanescent-wave illumination mode. Starting from the point that every objective exceeding a certain numerical aperture is generating evanescent waves, we demonstrate how the optical path can be modified to suppress the far-field excitation, and how one can switch easily between these types of illumination. The phenomena resulting from interactions of evanescent waves with cells are reviewed. The ways in which systematic variations of the angle of incident light can be used to obtain quantitative information on fluorophore distance, distribution and concentration are also discussed. PMID- 11482814 TI - The effects of a diode laser (810 nm) on pigmented guinea-pig skin. AB - Various types of lasers, such as the Q-switched ruby laser and the Alexandrite laser, cause selective damage to cutaneous pigmented cells and are currently used in the therapy of pigmented lesions. The aim of this study was to evaluate the effects of a diode laser at a wavelength of 810 nm on pigmented guinea-pig skin. The diode laser was supplied by OcuLight Iris Medical Instruments, Inc. and was used to deliver 0.15, 0.2, 0.3, 0.4, 0.5, 1 and 1.5 J/cm2 laser beams in micropulses of 100 micros. The study was carried out on albino and black-spotted guinea pigs (GP). After irradiation, punch biopsies were taken and analysed by light and electron microscopy. Albino animals developed just a few signs of cutaneous injury. This mostly consisted of spongiotic disarray, after the highest doses were administered (>1 J/cm2). In the black skin of spotted GPs alterations appeared at 0.15 J/cm2 and included melanosome damage and, at doses higher than 0.3 J/cm2, also melanocyte damage. These observations demonstrate that the diode laser at 810 nm selectively affects pigmented structures and that the cellular targets of diode laser radiation are the melanosomes. The diode laser specificity for melanin may provide a biological basis for the treatment of pigmented superficial cutaneous lesions. PMID- 11482815 TI - Monitoring of the infrared radiation emitted from skin layers during CO2 laser resurfacing: a possible basis for a depth navigation device. AB - CO2 laser resurfacing is very accurate, but it is not free of complications such as scarring. The objectives of the present study were to evaluate the pattern of the infrared (IR) radiation emitted from skin layers and to use this pattern to distinguish between these layers during resurfacing. A CO2-resurfacing laser (Sharplan SilkTouch) was used for the de-epithelialisation of skin. A silver halide optical fibre delivered the radiation emitted from the skin during resurfacing to an IR photonic detector. Time-dependent curves of the signals emitted from the skin layers were statistically evaluated and showed significant differences between the epidermis and the dermal layers. Similar results were obtained during in-vivo and ex-vivo measurements. The difference between the skin layers emission may be used for depth navigation during laser resurfacing. PMID- 11482816 TI - Laser doppler flowmetry in combined needle acupuncture and moxibustion: a pilot study in healthy adults. AB - Laser Doppler flowmetry (LDF) (DRT 4/Moor Instruments Ltd, Devon, UK) was used in this pilot study for monitoring the effects of an invariable acupuncture pattern on microcirculation of the skin before, during, and after combined needle acupuncture and moxibustion in 12 healthy volunteers (mean age 35.2 +/- 4.4 years, range 26-41 years, four female and eight male). According to the standards of traditional Chinese medicine (TCM), this acupuncture pattern is assumed to be unspecific and non-therapeutic. Flux decreased during the treatment period (p<0.05) compared to the control phase before combined needle acupuncture and moxibustion. After the removal of the needles, flux did not return to the initial control value. No significant differences between mean arterial blood pressure, heart rate, skin temperature or gender were detected. A prediction of individual incidences and kinds of acupuncture effects in our healthy volunteers was not possible. Modern monitoring techniques like LDF could be a method to separate responders from non-responders to acupuncture in peripheral microcirculatory disorders. Further studies on patients with peripheral microcirculatory disorders are necessary in order to demonstrate the value of LDF in detecting responders/non-responders in combination with therapeutic acupuncture patterns according to TCM. PMID- 11482817 TI - In vivo expression of the collagen-related heat shock protein HSP47, following hyperthermia or photodynamic therapy. AB - Heat shock protein 47 (HSP 47), a molecule expressed constitutively in cells that synthesise collagen, is involved in collagen type I biosynthesis, and after insult acts as a stress response molecule to sequester abnormal procollagen. Photodynamic therapy (PDT) is claimed not to result in extensive collagen damage, such as that which can occur after other laser treatments, e.g. hyperthermia (HT) or coagulation, thereby conferring on PDT a potential therapeutic advantage. In previous studies on mouse fibroblasts in vitro we demonstrated HSP47 elevation in the first hours after the application of conditions known to damage collagen, and an absence of HSP47 elevation following PDT with two well-established photosensitisers, haematoporphyrin ester (HpE) and meta-tetrahydroxyphenylchlorin (mTHPC). The present study examines HSP47 metabolism in murine skin following (1) HT, (2) PDT with HpE and (3) PDT with riboflavin (RB). Riboflavin was examined because of reports of collagen injury induced by its photoactivation. All three stresses were applied at grossly equitoxic, 'tolerance' doses. Three months after these doses, linear extensometry revealed the skin to have fibrotic characteristics after HT and RB PDT, but not after HpE PDT. HSP47 expression levels were analysed at transcriptional (Northern) and translational (Western) levels at early time intervals up to 24 h after the treatment application, starting immediately after the treatment for mRNA and 6 h post-treatment for protein. Highly significant upregulation of HSP47 was detected following HT, and PDT with RB. PDT mediated by HpE did not have any impact on HSP47 levels. These results were thus consistent with those from in vitro work and support the hypothesis of early elevation of HSP47 expression only by modalities affecting collagen or its precursors. PMID- 11482818 TI - An affordable, portable fluorescence imaging device for skin lesion detection using a dual wavelength approach for image contrast enhancement and aminolaevulinic acid-induced protoporphyrin IX. Part I. Design, spectral and spatial characteristics. AB - Steady-state fluorescence imaging can be used in conjunction with selective exogenous or endogenous fluorescent compounds for the diagnosis of skin lesions, for example cancerous lesions. Depending on the excitation and emission properties of the fluorescent compound used, various excitation and/or emission wavelengths can be chosen in order to allow fluorescence imaging. Unwanted background signals like autofluorescence and scattering can decrease the image quality and hence the diagnosis potential of this imaging method. A method involving two excitation and/or emission wavelengths was used in order to suppress the unwanted background signal and allow contrast enhanced fluorescence imaging. A fluorescence imaging device prototype was assembled using both the two wavelength excitation method and the two wavelength emission method. Additionally, a white light source was included to allow the collection of images as seen with the naked eye. The prototype was designed to be affordable and portable and was laid out towards the diagnosis of skin lesions using aminolaevulinic acid-induced protoporphyrin IX (PpIX). This paper describes the excitation and detection characteristics of a fluorescence imaging device prototype. This includes spectral and spatial characteristics of the various light sources included in the device as well as specifications of the image detector used. Furthermore, the image analysis procedure used for the dual wavelength excitation/emission is described. PMID- 11482819 TI - An affordable, portable fluorescence imaging device for skin lesion detection using a dual wavelength approach for image contrast enhancement and aminolaevulinic acid-induced protoporphyrin IX. Part II. In vivo testing. AB - A fluorescence imaging prototype for skin lesion detection and diagnosis using aminolaevulinic acid (ALA) induced protoporphyrin IX (PpIX) was tested in vivo and in the clinic. The prototype was designed as an affordable, portable device to allow contrast enhanced imaging of skin lesions using either the dual excitation wavelength method or the dual emission wavelength method or both. In this study the prototype was tested first on an animal model. Topical application of ALA on defined spots on mouse skin gave PpIX fluorescence after about 3 hours of application. After successful in vivo testing the instrument was tested on basal cell carcinoma patients before ALA-PpIX photodynamic therapy. The patients were topically applied with ALA. After three hours the device was tested (immediately before treatment). The prototype showed good results in terms of contrast enhancement (elimination of unwanted background signals, e.g. autofluorescence) using either contrast enhancement method, both methods achieving similar results. The results achieved in this study, combined with the affordable design of the device, seem to allow cost-effective, contrast-enhanced imaging of skin lesions before or during photodynamic therapy using ALA induced PpIX. PMID- 11482820 TI - Stimulation of MCM3 gene expression in osteoblast by low level laser irradiation. AB - Biostimulatory effect of cell proliferation and bone formation by laser irradiation has been reported, however, very little is known about the molecular basis of mechanisms. We previously constructed the cDNA library of mouse osteoblastic cells (MC3T3-E1) which enhanced gene expression by laser irradiation using a subtracted gene cloning procedure. In the present study, we focused on a gene clone, designated as MCL-140, which exhibited the high homology of DNA sequence with mouse minichromosome maintenance (MCM) 3 gene. MCM3 is involved in the initiation of DNA replication as licensing factor in eukaryotic cells. Nucleotide sequence of MCL-140 insert was determined and assessed in the nucleic acid databases. The transcription level of MCL-140 was examined by Northern blot analysis. The DNA sequences of clone MCL-140 insert exhibited 96.2% homology with MCM 3 gene coding P1 protein. Higher MCM3 mRNA levels were observed in laser irradiated cells compared to the levels in non-irradiated cells: furthermore, radiolabelled thymidine incorporation was increased by laser irradiation. These findings suggest that low-level laser irradiation may enhance DNA replication and play a role in stimulating proliferation of osteoblast through the enhancement of the MCM3 gene expression. PMID- 11482821 TI - Inhibition of interleukin-1beta production and gene expression in human gingival fibroblasts by low-energy laser irradiation. AB - Human gingival fibroblast (hGF) cells reside in gingival tissues which are challenged frequently by oral bacteria. Lipopolysaccharide (LPS) from periodontal pathogens can penetrate gingival tissues and stimulate the production of interleukin-1beta (IL-1beta), which has been implicated in inflammation and bone resorption. The anti-inflammatory effects of low-energy laser irradiation have been reported, but the mechanisms of this biostimulatory effect have not been fully elucidated. Primary cultured hGF cells were challenged with LPS isolated from Campylobacter rectus, a known periodontal disease-associated pathogen, and irradiated by a Ga-Al-As diode low-energy laser (830 nm, 3.95-7.90 J/cm2). The hGF cells cultured medium showed a marked elevation of IL-1beta production by LPS, which was significantly inhibited by laser irradiation in a dose-dependent manner. By reverse transcription-polymerase chain reaction (RT-PCR) analysis, this inhibitory effect was involved in the reduction of IL-1beta mRNA levels but not that of the IL-1beta converting enzyme. PMID- 11482822 TI - Does low penetration of human skin by the normal mode ruby laser account for poor permanent depilatory success rates? AB - Studies reported to date have shown a good depilatory response from patients treated with the normal mode ruby laser (NMRL) over 12 weeks, but a low response over a time period greater than this. Previous publications have suggested that this could be accounted for by the apparently poor skin penetration of laser light and so this study attempted to assess whether this was indeed the case. Skin samples of varying thicknesses were taken from six Caucasian patients and their depths measured. Each was laid individually on an energy meter before having pulses from an NMRL compatible with clinical doses (4.75 J/cm2, 9.24 J/cm2 and 13.41 J/cm2) fired on the epidermis. Several samples had the laser fired repetitively on the surface to assess whether this caused any change in laser/skin fluence depth profiles. Repetitive firing of the NMRL on the epidermis of skin samples did not alter the energy recorded by the meter beneath. The fluence/depth profiles were constructed showing the majority of energy was lost within the first 1 mm of the skin surface (50%) which then further reduced over distance but at a much slower rate. The maximum depth of penetration was 14.8 mm (SD +/- 0.478) which appeared to be a function of wavelength and not fluence. The results suggest that laser penetration of skin should be adequate for generating enough heat at the hair bulge and bulb, potentially causing permanent damage. The implications of this study are that it is probably the presence of the correct chromophore in large enough amounts which is required for successful permanent depilation to occur. PMID- 11482823 TI - Standing on the shoulders... PMID- 11482824 TI - Coast to Coast and Dr. Marbach. PMID- 11482825 TI - Dr. Melis comments on Raigrodski, et al.'s article in the January 2001 issue of CRANIO. PMID- 11482826 TI - The trigeminal nerve. Part IV: the mandibular division. AB - The mandibular or third division of the trigeminal nerve is the largest of the three divisions. It is considered a mixed nerve. That is, like the ophthalmic and maxillary divisions, the mandibular conveys afferent fibers. But unlike the former two divisions, the mandibular also contains motor or efferent fibers to the muscles of mastication, the mylohyoid and anterior digastric muscles, and the tensor veli palatini and tensor tympani muscles. So intimately associated with dentistry, the mandibular nerve has also been termed the dental nerve by anatomists in the past. This extensive and complicated division of the trigeminal nerve can cause confusion to both patient and doctor. Pain is often referred within its branches and even into other trigeminal divisions, chiefly the maxillary. This fourth and last article about the trigeminal nerve will present in detail the mandibular division. PMID- 11482827 TI - Electromyographic evidence of reduced muscle activity when ULF-TENS is applied to the Vth and VIIth cranial nerves. AB - The object of this study was to determine if the resting muscle activity of TMD patients with measured hyperactivity (EMGave > 2.0 microvolts) could be reduced by the application of ULF-TENS (Ultra-Low-Frequency-Transcutaneous Electrical Neural Stimulation) [(BioTENS) BioResearch, Inc. Milwaukee, WI]. Twenty-nine patients with resting muscle hyperactivity and ten without resting muscle hyperactivity were selected from a series of 124 consecutively diagnosed TMD patients. Electromyographic records were taken bilaterally (with the mandible at rest) from the superficial masseter, anterior temporalis, anterior digastric, and posterior temporalis muscles before and after the application of ULF-TENS. The EMG data were averaged. For the 85 muscles that were found initially to exceed the usual cutoff of 2.0 microvolts, means and standard deviations were calculated. The "before TENS" levels were: mean = 3.353 +/- 1.44227 and the "after TENS" values were: mean = 1.844 +/- 0.92421. Using a student t-test, we found the difference between the before and after levels significant (p < .0005). Additionally, means and standard deviations were calculated for each muscle (Ta, Mm, etc.) separately and also found to be significantly different (before Vs after TENS). In these "hyperactive" patients, the nonhyperactive muscles were also pooled, the means calculated before (mean = 1.21 +/- 0.456) and after (mean = 1.00 +/- 0.345) TENS and found to be significantly different (p < 0.01). For the ten patients exhibiting no muscle exceeding the 2.0 microvolt cutoff (mean = 1.08 +/- 0.415), no significant change (in levels) was observed after ULF-TENS (mean = 0.96 +/- 0.359; p > 0.88); however, when we calculated the "paired difference," we found a significant reduction, albeit very slight, in these patients' muscles, too. We conclude that ULF-TENS has an activity-reducing effect on the resting EMG levels of both hyperactive and relaxed muscles. It also appears that while the > 2.0 microvolt cutoff is useful for identifying patients that are hyperactive at rest, it does not identify complete relaxation of masticatory muscles. PMID- 11482828 TI - Fibrous adhesions in the joint compartment of the temporomandibular joint associated with experimental opposite drawing mandibular ramus in rabbit. AB - The purpose of this study was to develop a rabbit model of fibrous adhesions in the joint compartment of the temporomandibular joint (TMJ) without surgical invasion into the joint compartment. A unilaterally elastic force of about 120 grams was applied from the anterior edge of masseteric tuberosity to the posterior part of the zygomatic arch between the masseteric fascia layer and the subcutaneous layer in 14 rabbits. The blocks of nondrawing side TMJ were removed after four weeks. Fibrous adhesions were observed in 13 rabbits except for one rabbit that lost over 30% of its body weight. In eight of the rabbits, the adhesions were detected in both joint compartments. This animal model for fibrous adhesion of the TMJ will be useful in understanding the pathogenesis of adhesion in human TMJs. PMID- 11482829 TI - Histochemical studies of the masseter, the temporal and small zygomaticomandibular, and the temporomandibular masticatory muscles from aged male and female humans. Fiber types and myosin isoforms. AB - The purpose of this study was to investigate the histology of two small masticatory muscles from females and males of more than 70 years of age. By using immuno- and enzyme histochemistry the muscles were characterized by their fiber types and myosin heavy chain pattern. The observations were compared with similar studies of the masseter and temporalis muscles. Previously the two small muscles have been described based solely upon their gross anatomy. One muscle originates from the anterior, deep surface of the temporal fascia and inserts in the temporal tendon: the temporo-mandibular muscle (TM). The other muscle originates from the upper part of the temporal surface of the frontal process of the zygomatic bone and the adjacent part of the frontal bone and inserts in the temporal tendon: the zygomaticomandibular muscle (ZM). In the masseter, TM, and ZM, most of the autopsy samples contained an abundant number of fibers containing neonatal myosin heavy chains while in the temporal muscle specimens, such fibers were sparse and scattered. Electrophoresis followed by immuno-staining of Western blots supported the histochemical findings. There was no obvious correspondence between fiber typing based upon ATPase activity and the neonatal myosin heavy chain content in the muscle fibers. Neither did the fibers show accordance in their content of adult slow and fast myosin heavy chains and in their content of neonatal myosin heavy chain. PMID- 11482830 TI - Association between symptoms of temporomandibular disorders and depression: an epidemiological study of the Northern Finland 1966 Birth Cohort. AB - Facial pain and other symptoms of temporomandibular disorders (TMD) are rather common in the adult population. According to clinical studies, psychological factors play an important role in the etiology and maintenance of these symptoms. On the other hand, chronic pain can cause depression. The aim of this study was to evaluate the association between symptoms of TMD and depression in a large population sample of young adults. The study was a part of the 31-year follow-up study of the Northern Finland Birth Cohort consisting of 12,058 live births from the year 1966. Questionnaire information concerning TMD symptoms was collected from a subsample of 5,696 subjects. Depression was measured with a question about reported depression (diagnosed by a doctor) and with the Symptom Checklist depression subscale (SCL-25 DS). Of the TMD symptoms, those related to pain had the most significant relations to indicators of depression. In both genders, the proportion of depression indicated with the SCL-25 DS was significantly higher in subjects with pain-related symptoms of TMD, i.e., facial pain and "pain at jaw rest", and in men with "pain on jaw movement", compared with non-pain subjects (p<0.05). Other symptoms of TMD also associated significantly with SCL-25 DS (p>0.05), except "difficulties in mouth opening" among women. Among women, the prevalence of recognized depression was also significantly higher in subjects with pain-related symptoms of TMD, compared with subjects with no pain (p< or =0.05). Almost all the associations remained significant after adjusting for marital status, education, and self-rated general health. In conclusion, the results show that depression has an association with TMD symptoms, especially those related to pain. When treating patients with facial pain, dentists should consider the possible presence of psychopathology and, if necessary, consult appropriate mental health professionals. PMID- 11482831 TI - Temporomandibular disorders--traumatic etiology vs. nontraumatic etiology: a clinical and methodological inquiry into symptomatology and treatment outcomes. AB - The purpose of this research is to investigate the distinctions relating to Presenting Symptoms and Treatment Outcomes between patients suffering temporomandibular disorder (TMD) as a result of traumatic versus nontraumatic etiology. A geographically diverse cohort of 1,842 patients diagnosed with TMD was investigated with special emphasis placed on the following criteria: 1. The distribution of demographic and symptom characteristics of patients with trauma as an immediate precipitating factor versus those with other nontraumatic etiologies; 2. The relationship between nontrauma status and treatment outcomes; 3. The interrelationships between nontrauma status, psychosocial factors, and treatment outcomes. Trauma patients tended to be younger, less educated, and more likely to be male than the nontrauma patients. For this group the reported length of the TMD problem was, as expected, of shorter duration when compared to the nontrauma patient group. Length of treatment did not differ between the two groups. In comparison, trauma patients reported higher initial overall symptoms including pain and range of motion limitations. However, symptoms related to joint dysfunction did not vary appreciably. Treatment outcomes are complicated by the fact that TMD encompasses several different diagnostic entities. Trauma patients reported significantly higher percentages of improvement in palpation pain and perceived malocclusion. No significant differences were found for pain report, joint dysfunction, stress, and overall TMD symptomatology, as measured by the TMJ Scale's Global domain. Trauma patients manifested higher psychological dysfunction levels (excepting stress) and showed significantly more improvement in both psychosocial function and stress than the nontrauma group. PMID- 11482832 TI - Orthodontic finalization strategies in dysfunctional adult patients. AB - Although orthodontic treatment for the finalization of temporomandibular dysfunction (TMD) patients is often advocated, it represents a risk of failure. Furthermore, in the literature there are few articles that illustrate appropriate orthodontic strategies for the finalization of these patients. In this article the authors suggest a step-by-step clinical procedure in order to: 1. verify the rationale for orthodontic finalization in TMD patients; 2. illustrate the guidelines for correct orthodontic treatment; and 3. suggest a strategy to stabilize the mandible during orthodontic treatment and to retain that stabilization after treatment. If splint therapy is successful in relieving the TMD symptoms, a fixed posterior bilateral stop is used for occlusal stability. After one or two months during which the bilateral stop is shown to be adequate in maintaining TMJ health without a splint, orthodontic finalization can be performed with minimal risk of failure. The essential sequences of Class I, Class II, and Class III treatment for the finalization of TMD patients are also presented. The difficult goal of a long term stability in cases that have been orthodontically finalized is reached by the conversion of the posterior bilateral stops (crown build-ups) into permanent vertical stops. PMID- 11482833 TI - Handling difficult people and difficult situations. PMID- 11482834 TI - Developmental functions for saccadic eye movement parameters derived from pro- and antisaccade tasks. AB - The few studies on the development of oculomotor functions conducted so far suggest strong developmental effects of age on different parameters of saccade control in childhood and adolescence. The present study aimed at determining developmental functions for a set of 22 parameters derivable from the examination of pro- and antisaccades elicited under the 200-ms gap and overlap conditions (100 trials under each of the four conditions). The statistical analyses (including multiple regression, analysis of covariance, and principal components analysis (PCA) were based on a sample of 199 subjects aged 6-28 years, including 66% males and 34% females. We obtained the following main results: (1) In most cases, the variable "age(-1)" predicted the dependent variables much better than age. (2) Multiple regression approaches using Age and Age(-1) as predictors accounted for 0-51% of the criterion variances, these values (adjusted R2) being large for the proportion of direction errors during the antisaccade tasks (0.46 0.51), medium for anti- (0.36-0.46) and prosaccadic (0.23-0.34) reaction times (RT) and their standard deviations (0.26-0.45), and negligible for the proportion of express saccades. (3) The age variables (particularly Age(-1)), furthermore, predicted some of the effects of the experimental task manipulations on the different dependent variables (e.g. the augmentation of direction errors under the gap as compared to the overlap condition of the antisaccade task) significantly. (4) PCA results suggest 5 factors of saccade control in this sample: 2 factors comprising the antisaccade parameters, 2 factors comprising the prosaccade parameters, and 1 factor comprising the anticipations. The best correlations of the age variables with the corresponding factor scores were obtained for the 2 factors related to the antisaccade task parameters. PMID- 11482835 TI - Symmetric facilitation between motor cortices during contraction of ipsilateral hand muscles. AB - Using transcranial magnetic stimulation (TMS) over the contralateral motor cortex, motor evoked potentials (MEPs) were recorded from resting abductor pollicis brevis (APB) and first dorsal interosseous (FDI) muscles of eight subjects while they either rested or produced one of six levels of force with the APB ipsilateral to the TMS. F-waves were recorded from each APB at rest in response to median nerve stimulation while subjects either rested or produced one of two levels of force with their contralateral APB. Contraction of the APB ipsilateral to TMS produced facilitation of the MEPs recorded from resting APB and FDI muscles contralateral to TMS but did not modulate F-wave amplitude. Negligible asymmetries in MEP facilitation were observed between dominant and subdominant hands. These results suggest that facilitation arising from isometric contraction of ipsilateral hand muscles occurs primarily at supraspinal levels, and this occurs symmetrically between dominant and subdominant hemispheres. PMID- 11482836 TI - Critical flicker frequency responses in visual cortex. AB - Critical flicker frequency (CFF) threshold is defined as the frequency at which a flickering light is indistinguishable from a steady, non-flickering light. CFF is useful for assessing the temporal characteristics of the visual system. While CFF responses are believed to reflect activity in the central visual system, little is known about how these temporal frequencies are processed in the visual cortex. The current paper estimated the CFF threshold for cells in the rat visual cortex by recording single unit responses to flickering stimuli. Results showed that: (1) there was a broad range of temporal tuning, (2) CFF threshold was lower in simple cells than in complex and hypercomplex cells, and (3) there was no significant difference in CFF threshold between areas 17 and 18. PMID- 11482837 TI - Responses of rostral fastigial neurons to linear acceleration in an alert monkey. AB - Vestibular-only neuronal responses to angular acceleration have been systematically characterized in the rostral fastigial nucleus (FN) by several studies. However, responses of these neurons to linear acceleration have not been examined. In this study, we recorded single-unit activity of vestibular-only neurons in an alert monkey during pure sinusoidal linear acceleration along different directions in the horizontal plane. Spatiotemporal response properties were quantified by computing two-dimensional response ellipses in the horizontal plane. Based on this analysis, neurons were classified as narrowly or broadly tuned. About 29% (5/17) of neurons were broadly tuned. The other 71% (12/17) were narrowly tuned. Unlike vestibular nuclei neurons, all recorded FN neurons exhibited irregular resting discharge rates (CV*0.2). Based on studies of linear motion-sensitive neurons in the vestibular nuclei, the data suggest that irregular neurons in the rostral FN and the vestibular nuclei have similar responses to linear acceleration in behaving monkeys. PMID- 11482838 TI - Neuronal activity in dorsomedial frontal cortex and prefrontal cortex reflecting irrelevant stimulus dimensions. AB - Previous studies of the dorsomedial frontal cortex (DMF) and the prefrontal cortex (PF) have shown that, when monkeys respond to nonspatial features of a discriminative stimulus (e.g., color) and the stimulus appears at a place unrelated to the movement target, neurons nevertheless encode stimulus location. This observation could support the idea that these neurons always encode stimulus location, regardless of its relevance to an instrumentally conditioned behavior. Past studies, however, leave open the possibility that activity observed during one operant task might reflect the contingencies of a different task, performed at different times. To test these alternatives, we examined the activity of DMF and PF neurons in two rhesus monkeys conditioned to perform an operant eye movement task in which only the color and shape of visual stimuli served as salient discriminative features. Each of eight stimuli was associated with a response to a different eye-movement target. The location of these stimuli varied from trial to trial but was of no behavioral relevance, and the monkeys did not perform any operant task in which stimulus location controlled behavior. A substantial minority of neurons in both DMF and PF nevertheless encoded stimulus location, which indicates that this property does not depend on its relevance in an instrumentally conditioned behavior. PMID- 11482839 TI - Postural coordination patterns associated with the swinging frequency of arms. AB - Voluntary arm movements frequently perturb body equilibrium in an upright posture. The motions of leg joints need to be coordinated according to the properties of voluntary arm movements in order to maintain body equilibrium, and this may cause a change in postural pattern. The purpose of this study was to determine whether the kinematic pattern generation of upright posture is influenced by a change in the swinging frequency of arm movements and whether the pattern generation is correlated with a change in joint torque about the shoulder joint. Four male subjects in an upright posture were instructed to swing their arms at seven different frequencies, determined by the maximum swinging frequency of each subject (35%max, 40-60%max, 65%max). Segment rotations around the shoulder, hip, and ankle joints were analyzed at kinematic and kinetic levels. The results of kinematic analysis indicated that tight coupling between motions of the shoulder and hip joints was generated in lower-frequency trials (under 40 45%max), whereas tight coupling between motions of the shoulder and ankle joints was generated in higher frequency trials (more than 40-45%max). Furthermore, the results of kinetic analysis revealed that changes in the joint torque patterns about the shoulder and hip joints occurred in trials at 40-45%max. The mean value of 40-45%max was close to the eigenfrequency of each subject's arm. We concluded that (1) postural patterns associated with a gradual change in the swinging frequency of the arms can be divided into two coordination modes (a hip-shoulder in-phase mode and an ankle-shoulder in-phase mode), and (2) these two patterns may be divided by the eigenfrequency of the arm. PMID- 11482840 TI - Neural compensation for mechanical loading of the hand during coupled oscillations of the hand and foot. AB - The role of kinaesthetic afferences in controlling coupling of voluntary oscillation of the hand and foot, both in-phase and anti-phase, was investigated by modifying the mechanical properties of one of the two segments (the hand) with applied inertial or elastic loads. Loads consisted of a lead disk, rotating coaxially with the wrist (total inertial momentum 15 g m2), or in two symmetrical rubber bands (elasticity, 4 g deg(-1)) connected 5 cm away from the wrist pivot. Experiments were performed on five male and five female subjects. Both the frequency responses of the hand and foot (i.e. the phase relations between the onset of muscular activation in limb extensors and the onset of the related movement) and the inter-limb phase relations (the phase differences between the hand and foot movement cycles and between the onsets of the electromyographic (EMG) activity in hand and foot extensors) were analysed. The hand frequency response was fitted with a 2nd-order model, allowing us to describe the loaded and unloaded conditions through the changes in the model response. Inertial loading induced an immediate and steep decay in the frequency response, with a clear-cut reduction of the model resonance frequency, while elastic loading shifted the response to the right and upwards. Inter-limb phase relations were only partially affected by inertial loading of the hand. Despite the fact that the load strongly increased the difference between the frequency-responses of the hand and foot, when hand and foot were oscillated in-phase only about half of this difference remained as an increased phase-lag between hand and foot oscillations. The other half was offset by an advance of the contraction of the hand movers with respect to the foot movers. This compensation mechanism was more effective during anti-phase than during in-phase movements. Elastic loading improved inter-limb synchronisation, since it superimposed the hand frequency response on that of the foot. In this condition, the requested synchronisation (in-phase or anti-phase) could be achieved by an almost simultaneous (or in strict phase opposition) contraction of the hand and foot movers. In conclusion, the main feedback reaction to the de-coupling effect of hand loading consisted in modifying the timing of activation of the muscles moving the extremities. An advance of hand movers on foot movers is already present in unloaded conditions to compensate for the difference in the natural mechanical properties of the two segments. This advance is enhanced when increasing the inertia of the hand system and attenuated when increasing its elasticity. PMID- 11482841 TI - Fictive motor activity in rat after 14 days of hindlimb unloading. AB - The goal of the present study was to examine the effects of chronic hindlimb unloading on fictive motor patterns which can be developed in hindlimb nerves of adult rats. The animals were divided into two groups. The first group was submitted to hindlimb unloading for 2 weeks by tail suspension. The second group served as controls. After this initial phase, the animals of both groups were acutely decorticated, paralysed and electroneurographic efferent activity was recorded from hindlimb muscle nerves under conditions of "fictive locomotion" in order to evaluate variations in central locomotor command. Fictive rhythmic motor episodes were either spontaneous or evoked by electrical stimulation of the mesencephalic locomotor region. Only the second ones were recognised as locomotor like activities. The motor pattern was not fundamentally affected by unloading except that, after the unloading period, extensor muscle nerves were significantly more frequently activated and their burst durations were increased compared to activity in control animals, despite the fact that the phasic sensory afferent inputs were suppressed. This suggests that unloading induces plastic modifications of the central networks of neurons implicated in the locomotor command. The origin of this extensor hyperactivity is discussed. It is proposed that it could be the consequence of either changes in motoneuronal properties or of an increase in afferent input to motoneurones. PMID- 11482842 TI - The ventral hippocampus and fear conditioning in rats. Different anterograde amnesias of fear after tetrodotoxin inactivation and infusion of the GABA(A) agonist muscimol. AB - Studies on the involvement of the rat hippocampus in classical fear conditioning have focused mainly on the dorsal hippocampus and conditioning to a context. However, the ventral hippocampus has intimate connections with the amygdala and the nucleus accumbens, which are involved in classical fear conditioning to explicit and contextual cues. Consistently, a few recent lesion studies have indicated a role for the ventral hippocampus in classical fear conditioning to explicit and contextual cues. The present study examined whether neuronal activity within the ventral hippocampus is important for the formation of fear memory to explicit and contextual cues by classical fear conditioning. Tetrodotoxin (TTX; 10 ng/side), which completely blocks neuronal activity, or muscimol (1 microg/side), which increases GABA(A) receptor-mediated inhibition, were bilaterally infused into the ventral hippocampus of Wistar rats before the conditioning session of a classical fear-conditioning experiment. Conditioning to a tone and the context were assessed using freezing as a measure of conditioned fear. TTX blocked fear conditioning to both tone and context. Muscimol only blocked fear conditioning to the context. The data of the present study indicate that activity of neurons in the ventral hippocampus is necessary for the formation of fear memory to both explicit and contextual cues and that neurons in the ventral hippocampus that bear the GABA(A) receptor are important for the formation of fear conditioning to a context. In addition, both bilateral muscimol (0.5 microg/side and 1 microg/side) and TTX (5 ng/side and 10 ng/side) infusion into the ventral hippocampus dose-dependently decreased locomotor activity in an open-field experiment. PMID- 11482843 TI - Pre-excitatory pause in frontal eye field responses. AB - We report a new characteristic of the presaccadic activity of the neurons in the frontal eye field of macaque monkeys. A fraction of neurons exhibited a significant pause in discharge rate preceding the excitatory visual or movement related response. This pre-excitatory pause, which has been observed in striate and extrastriate visual areas, may represent a resetting of neural activation for detailed visual processing or saccade preparation. PMID- 11482844 TI - Systematic errors of planar arm movements provide evidence for space categorization effects and interaction of multiple frames of reference. AB - Healthy humans performed arm movements in a horizontal plane, from an initial position toward remembered targets, while the movement and the targets were projected on a vertical computer monitor. We analyzed the mean error of movement endpoints and we observed two distinct systematic error patterns. The first pattern resulted in the clustering of movement endpoints toward the diagonals of the four quadrants of an imaginary circular area encompassing all target locations (oblique effect). The second pattern resulted in a tendency of movement endpoints to be closer to the body or equivalently lower than the actual target positions on the computer monitor (y-effect). Both these patterns of systematic error increased in magnitude when a time delay was imposed between target presentation and initiation of movement. In addition, the presence of a stable visual cue in the vicinity of some targets imposed a novel pattern of systematic errors, including minimal errors near the cue and a tendency for other movement endpoints within the cue quadrant to err away from the cue location. A pattern of systematic errors similar to the oblique effect has already been reported in the literature and is attributed to the subject's conceptual categorization of space. Given the properties of the errors in the present work, we discuss the possibility that such conceptual effects could be reflected in a broad variety of visuomotor tasks. Our results also provide insight into the problem of reference frames used in the execution of these aiming movements. Thus, the oblique effect could reflect a hand-centered reference frame while the y-effect could reflect a body or eye-centered reference frame. The presence of the stable visual cue may impose an additional cue-centered (allocentric) reference frame. PMID- 11482845 TI - Saturable brain-to-blood transport of endomorphins. AB - Opiate-modulating tetrapeptides such as tyrosine-melanocyte-stimulating hormone release inhibiting factor-1 (Tyr-MIF-1; Tyr-Pro-Leu-Gly-NH2) and Tyr-W-MIF-1 (Tyr Pro-Trp-Gly-NH2) are saturably transported from brain to blood. We examined whether two recently described endogenous opiate tetrapeptides with similar structures, the mu-specific endomorphins, also are transported across the blood brain barrier (BBB). We found that the efflux rates of endomorphin-1 (Tyr-Pro-Trp Phe-NH2) and endomorphin-2 (Tyr-Pro-Phe-Phe-NH2) were each self-inhibited by an excess of the respective endomorphin, thereby defining saturable transport. Cross inhibition of the transport of each endomorphin by the other indicated shared transport. By contrast, no inhibition of the efflux of either endomorphin resulted from coadministration of Tyr-MIF-1, indicating that peptide transport system-1 (PTS-1) was not involved. Tyr-W-MIF-1, which is partially transported by PTS-1, significantly (P<0.01) decreased the transport of endomorphin-1 and tended (P=0.051) to decrease the transport of endomorphin-2, consistent with its role as both an opiate and antiopiate. Although involved in modulation of pain, coinjection of calcitonin gene-related peptide or constriction of the sciatic nerve did not appear to inhibit endomorphin efflux. Thus, the results demonstrate the existence of a new efflux system across the BBB which saturably transports endomorphins from brain to blood. PMID- 11482846 TI - Two kinds of body representation are used to control hand movements following tactile stimulation. AB - The task of reaching with a hand toward a target given in space is generally described as an ill-posed problem. It is often assumed that some kind of internal body model is required to solve this problem. This article provides information concerning the nature of this representation. Experiments were carried out in which blindfolded subjects were stimulated mechanically at one of eight possible stimulation sites on the legs and then asked to move one hand as quickly as possible to the stimulated site. In nine different postures of legs and hands, the frequency was recorded of the use of either hand. In addition, reaction times (RT) were measured and in two choice reaction tasks RTs were measured for different conditions (morphologically compatible vs morphologically incompatible, hands parallel vs hands crossed): The results support the hypothesis that the representation of the body is based on at least two systems, one which provides spatial information concerning the body position and a second one which is based on a morphological representation. According to this hypothesis, our results could be described by the following schema. Following the application of the stimulus, two processes were started in parallel. One concerned the activation of the spatial representation of body position, including the position of stimulus site and the possible response sites. This was more difficult, i.e., took more time, when the hands were crossed. Within this spatial representation, the distances between stimulus site and response sites were determined, and it was found that the smaller the distance, the more strongly the hands were activated. Simultaneously, in a second process the response site which was morphologically ipsilateral to the stimulus was excited. This schema could explain our results: In the behavioral experiments that hand which was most strongly excited, and which therefore exceeded a threshold sooner, won the decision and showed smaller RT values. In the choice reaction experiments, the winning hand was compared with the externally given task. The more strongly, according to the given task, the wrong hand was excited, the longer was the RT of the response. PMID- 11482847 TI - Independent movements of the digits in grasping. AB - Reaching out for an object is often considered to consist of the control of two components: transporting the hand to the object's position, and scaling the grip to the object's size. We recently proposed an alternative view. According to this view, grasping consists of controlling the digits, not the hypothetical transport and grip. This alternative view assumes that the opening of the hand emerges from the trajectories of the digits. We therefore studied the movements of the digits in grasping. We asked subjects to grasp disks (diameters ranging from 5 to 8 cm) at marked positions with two digits. The positions were at opposite sides of the disk, at the same distance from the starting position, so that the orientation of the surface was the same for both digits. The subjects grasped the disks either with the index finger and thumb of the dominant hand, with the same digits of the non-dominant hand, or bimanually with both index fingers. Our predictions are: that the well-known relation between object size and grip aperture holds for each digit; that the same relation holds if the object is grasped with two hands instead of with the thumb and finger of one hand; that maximum deviation, variability and duration of the digit movements are related; and that variations in the timing of the maximum deviation of one digit are independent of those in the other digit. In accordance with our predictions, we found that the maximum deviation of both digits increased with 0.75 times the object radius, independent of the hand(s) used. The movements of the thumb were more variable than those of the index finger, which was reflected by a larger deviation earlier in the movement. The timing of the maximum deviation of the two digits was independent. These results on the digits' movements are consistent with our view that grasping can be understood as the largely independent movements of the digits. The results are not in conflict with the hypothesis that the grip is controlled during grasping, but can only be explained by extending that hypothesis post hoc. PMID- 11482848 TI - The technology revolution: last gasps for the printed-journal Luddite? PMID- 11482849 TI - Disaster management, disaster medicine and emergency medicine. PMID- 11482850 TI - Asystolic cardiac arrest: poor outcome or terminal event? PMID- 11482851 TI - Complex humanitarian emergencies: a major global health challenge. AB - Complex humanitarian emergencies have been a major political, security and public health feature of the post-Cold War world. These man-made disasters account for more morbidity and mortality than all natural and technological disasters combined. In order to deliver effective aid during complex humanitarian emergencies, international relief agencies must have a solid understanding of the political and social climates in which they are operating. In addition, they should base their health interventions on objective epidemiological data, especially standardized rates of morbidity and mortality. Most deaths during complex humanitarian emergencies are due to preventable causes, especially increased rates of infectious diseases malnutrition and violent trauma. The most appropriate health interventions are therefore based on the models of public health and primary health care, emphasizing disease prevention and health promotion. The field of humanitarian assistance has become increasingly professionalized in recent years, with its own professional standards, literature, research agenda and training opportunities. It is an unfortunate reflection on the current state of international affairs that the number of complex humanitarian emergencies and the enormous levels of suffering associated with them are unlikely to decline in the foreseeable future. PMID- 11482852 TI - The CDC pacific emergency health initiative: a pilot study of emergency preparedness in Oceania. AB - OBJECTIVE: Environmental emergencies and disasters are becoming more frequent in developing nations. Between 1992 and 1996, disasters affected an annual average of 4.5 million Oceania residents. Unfortunately, public health planners in the region and responders throughout the world have little evidence on which to base measures of emergency preparedness. Indicators of preparedness must be identified, implemented and evaluated before the effectiveness of emergency planning interventions can be measured accurately. The aim of this study was to perform an objective evaluation of emergency preparedness among five nations in Oceania. METHODS: A standardized retrospective review of national-level public health and institutional-level hospital emergency operations plans from a convenience sample of five Pacific nations or territories was performed. In addition, in-country interviews, observation of operations and review of documentation were conducted. The rates of affirmative responses to 957 yes/no queries in the questionnaire were tabulated according to major emergency operational planning concepts and categories of emergency support functions. RESULTS: The study revealed remarkably low levels of emergency planning and preparedness among health and medical sectors of five Pacific islands. CONCLUSION: These data suggest a very low level of host national capacity for development of preparedness. Further investigation is necessary to define this need throughout this region of Oceania. PMID- 11482853 TI - Disaster management in Australia: the national emergency management system. AB - Australia has a well-established emergency management system to manage the threats to communities from natural and technological hazards. The key elements of the system are described with particular emphasis on the impact of disasters on Australia, fundamental concepts, and principles of planning. Emergency physicians have a key role in the development of emergency plans and in the overall health response to disasters. They can enhance their understanding of local, State and national arrangements by participating in exercises and appropriate disaster medicine and emergency management courses. PMID- 11482854 TI - Major Incident Medical Management and Support (MIMMS): a practical, multiple casualty, disaster-site training course for all Australian health care personnel. AB - Major Incident Medical Management and Support is a 3-day major incident training course designed specifically for doctors, nurses and ambulance personnel. It teaches a systematic, 'all hazards' approach to the principles of pre-hospital, multiple-casualty incident medical management. This article explores the origins and development of the course in Australia, it outlines the nature and content of the course, details the demographic profile of those who have successfully completed the course to date and, finally, looks at the future directions of the course in Australia. PMID- 11482855 TI - Reliability of modern microwave ovens to safely heat intravenous fluids for resuscitation. AB - OBJECTIVE: To determine if intravenous fluid heated in modern microwave ovens is warmed to a consistently safe temperature, as has been advocated in a number of texts and journals. METHODS: Five, 1L bags of normal saline in Viaflex (Baxter Healthcare, Old Toongabie, NSW, Australia) containers were heated for 2 min on high setting in 16 different microwave ovens. The output power ranged from 650 to 1,000 W. All microwave ovens were equipped with electronic timers and turntables. Initial and final temperatures were recorded with a TestoTerm 1100 electronic thermometer (Dade Behring, Lane Cove, NSW, Australia), accessing the centre of the fluid via the injection port. The average and standard deviation for each measurement was determined for each microwave oven. RESULTS: In nine out of the 16 microwaves (56%) tested, the fluid had reached an average final temperature greater than 42 degrees C and thus was greater than the recommended maximum temperature for the heating of intravenous fluids. All microwave ovens with an output power of greater than 900 W overheated the fluids. However 13/16 microwaves (81%) had a temperature range less than 3 degrees C, thus if correctly calibrated could be appropriate for heating intravenous fluid for resuscitation. CONCLUSIONS: The increased output power of modern microwave ovens can lead to overheating of resuscitation fluids, if the simple algorithm currently recommended is followed, leading to potentially serious complications. Microwave heating of intravenous fluid could be a safe, simple, cheap and effective means of heating intravenous fluids for resuscitation, but care needs to be taken to calibrate individual machines to ensure a safe temperature is reached. PMID- 11482856 TI - Asystolic cardiac arrest in Melbourne, Australia. AB - OBJECTIVE: To study the outcome of victims of out-of-hospital cardiac arrest presenting to the Metropolitan Ambulance Service in asystole, in Melbourne, Australia. METHODS: A retrospective case-note review of all patients presenting to the Metropolitan Ambulance Service in asystole for 1997 was performed. Metropolitan Ambulance Service case notes and hospital records were examined to determine the presenting rhythm and the patients' outcome. RESULTS: In a 12-month period, 778 patients met the entry criteria. Age mean was 67 years, 36% female, 64% male. Metropolitan Ambulance Service response time to scene time was a mean of 9.76 min. Resuscitation was commenced on 37% of patients. There was one survivor (0.12%). CONCLUSION: Adult victims of out-of-hospital cardiac arrest presenting as asystole should not receive treatment. PMID- 11482857 TI - Does routine use of the 15-lead ECG improve the diagnosis of acute myocardial infarction in patients with chest pain? AB - OBJECTIVE: It has been suggested that the use of additional electrocardiogram leads might improve the diagnostic sensitivity of this test, thus potentially expanding eligibility for thrombolysis for patients suffering myocardial infarction. The aims of this study were to evaluate the role of the 15-lead electrocardiogram in the emergency department chest pain population and to determine whether the routine use of the extra leads expands the group of patients eligible to receive thrombolysis. METHODS: Blinded, individual and independent analysis by two emergency physicians of paired 12- and 15-lead electrocardiograms from adult patients with a primary complaint of chest pain. The main outcome measure was the diagnosis of myocardial infarction eligible for thrombolysis. Data were analysed using descriptive statistics and kappa statistics for agreement between raters. RESULTS: 540 electrocardiograms (270 sets) were analysed. Myocardial infarction qualifying for thrombolysis was identified (by consensus) in 21 cases. In no case did the 15-lead electrocardiograph identify a myocardial infarction qualifying for thrombolysis that was not identified on the 12-lead electrocardiogram. CONCLUSION: In this study, the 15-lead electrocardiogram did not increase the number of thrombolysis eligible myocardial infarctions identified when compared with the 12-lead electrocardiogram. This study is limited by the small patient sample size, and a large multicentre trial is recommended to compare the 12- and 15-lead electrocardiograms in the emergency department population where the incidence of posterior and right-sided myocardial infarction is ultimately known. PMID- 11482858 TI - Same-day X-ray reporting is not needed in well-supervised emergency departments. AB - OBJECTIVE: To evaluate the efficacy of a missed radiological abnormality follow up system in a teaching hospital emergency department. METHODS: Prospective audit of all reported radiological abnormalities missed by Fremantle Hospital Emergency Department medical staff from 1 January 1997 to 31 December 1998. RESULTS: Of 29,724 radiological examination series, 459 abnormalities (1.5%) were not clearly documented as being identified in the medical record. The commonest missed abnormalities were incidental chest findings, distal wrist fractures with minimal or no displacement, radial head fractures and tibial plateau fractures. The most senior doctor undertaking initial film review was a junior medical officer in 242 cases (53%), a registrar in 96 cases (21%), and a consultant in 42 cases (9%). The most senior staff member was unknown in 79 cases (17%). One hundred and twenty-four missed abnormalities required a change in patient management (0.41% of total examinations, CI 0.34-0.48%). Ninety patients (73%) were referred to the patient's general practitioner for management. Seventeen patients (14%) returned to the emergency department for management. Thirteen patients (10%) were referred to a specialist clinic and in four cases (3%) the management of the patient was not recorded. No patient required re-admission to hospital. CONCLUSIONS: Missed radiological abnormalities in an emergency department with extended-hours emergency physician supervision can be managed non-urgently on an outpatient basis. Same-day reporting of radiographs is not required if adequate follow-up mechanisms for missed abnormalities exist. PMID- 11482859 TI - Improvement in decision-making for thrombolysis following introduction of angioplasty referral guidelines. AB - OBJECTIVE: To determine whether the introduction of acute angioplasty referral guidelines would lead to an improvement in decision-making times with respect to thrombolysis for patients with acute myocardial infarction. METHODS: Retrospective and prospective collection of 'time from diagnostic electrocardiogram to treatment' for all patients who received thrombolytic treatment for acute myocardial infarction in the emergency department. RESULTS: Median time from 'diagnostic electrocardiogram to thrombolysis' fell from 38 to 26 min (P < 0.0001) following introduction of acute angioplasty referral guidelines. This was primarily due to an improvement in median decision-making times for patients with anterior infarcts from 65 to 27 min (P = 0.0009). CONCLUSION: Significant improvements were seen in thrombolysis decision-making times for patients with acute myocardial infarction following the introduction of acute angioplasty referral guidelines. PMID- 11482860 TI - Plasma alkalinization for tricyclic antidepressant toxicity: a systematic review. AB - OBJECTIVE: To review the evidence that plasma alkalinization improves the outcome in tricyclic antidepressant toxicity. METHODS: Medline search from 1966 to October 2000 (articles in all languages were included) and examination of bibliographies. Published papers including animal studies, in vitro studies, human case reports, case series and retrospective studies were reviewed. RESULTS: Our search identified 115 publications, all of which were retrieved. Human studies included eight case reports, four case series, one controlled study and two retrospective chart reviews. No randomized controlled human trials were found. Twelve animal studies were identified that investigated pH manipulation or saline load and their effects on physiological parameters in tricyclic antidepressant toxicity. CONCLUSIONS: The practice of alkalinization for tricyclic antidepressant toxicity is based on animal studies, case reports and opinion. The mechanism of action appears to be multifaceted and may vary between different tricyclic antidepressants. Significant interspecies variation makes extrapolation from animal studies to humans difficult. Alkalinization therapy appears reasonable in patients with compromising dysrhythmias and shock when supportive interventions have been ineffective; however, the available evidence does not support prophylactic alkalinization in the absence of life-threatening cardiovascular toxicity. PMID- 11482861 TI - Management of venous thromboembolic disease in the lower limb. AB - Thromboembolic disease continues to cause significant morbidity and mortality in our community, despite extensive research into the aetiological factors and significant resources invested in the development of pharmacological agents for treating this condition. Development of more sensitive and specific modalities for identifying venous thromboses has improved their early detection, particularly in the commonest site, the lower limb. A rational evidence-based management pathway has not been formulated as debate continues over the most appropriate method of treatment. This review outlines the pathophysiology of the disease, provides a clinical pathway for the management of lower limb thromboembolic disease using reliable available evidence and briefly discusses the efficacy of drug therapy. PMID- 11482862 TI - Emergency Medical Services Fellowship in the United States of America. AB - Travelling outside Australia to undertake further training in an area of subspecialty interest is both interesting and beneficial to the advancement of the individual and our specialty. In the United States of America, such formal training following completion of specialist qualification in emergency medicine is referred to as 'Fellowship' training. While other authors have discussed the general areas of overseas work and emergency medicine Fellowships, this paper specifically addresses the area of prehospital care, known in the United States as 'emergency medical services'. Although there are significant differences in prehospital care between the United States and Australia, a great deal of what can be learned from undertaking a Fellowship in prehospital care in the United States is locally applicable. A typical curriculum is outlined, and the steps in selecting and arranging such a programme are discussed. Some potential pitfalls are also mentioned. Given the paucity of formal training in prehospital care in this country, such fellowship programmes are an excellent means of obtaining a very solid understanding of this important aspect of emergency medicine. PMID- 11482863 TI - Training for the role of triage in Australasia. AB - OBJECTIVE: To characterize the prerequisite experience and training undertaken by nurses for the role of triage in emergency departments in Australasia. METHODS: Postal survey of charge nurses/unit nurse managers of all Australasian emergency departments accredited for specialist emergency physician training by the Australasian College for Emergency Medicine. RESULTS: The response rate was 89%. The most common duration of prerequisite experience was 12-18 months. Most programmes use a combination of educational activities, with self-directed learning packages, lectures and mentored experience being the most common. Three hospitals reported no preparation for triage. CONCLUSION: In Australasia, there is wide variability in required training and experience before triage duties are performed. Strategies to set suggested minimum standards in these areas and to make training activities more accessible are recommended. PMID- 11482864 TI - Emergency doctors by sea to Antarctica: small ship medicine in polar regions. PMID- 11482865 TI - Fatal pneumococcal Waterhouse-Friderichsen syndrome. AB - There have only been six deaths reported in the English literature due to disseminated intravascular coagulation and Waterhouse-Friderichsen syndrome complicating pneumococcemia in otherwise healthy persons with normal spleens. Four of these deaths occurred in children and two in adults. A case of fulminant pneumococcemia complicated by disseminated intravascular coagulation and Waterhouse-Friderichsen syndrome in an otherwise healthy adult with a normal spleen is presented. The case is typical of the course of fulminant pneumococcal sepsis and highlights some of the difficulties experienced by rural general practitioners and rural retrieval services. Mechanisms by which Pneumococcus can elicit fulminant sepsis are discussed, although there is no explanation as to why this may occur in the setting of normal splenic function. PMID- 11482866 TI - Noxious gas exposure in the outback: two cases of hydrogen sulfide toxicity. AB - Two cases of toxicity from industrial exposure to hydrogen sulfide are presented. Both patients had neurological effects with loss of consciousness and amnesia along with respiratory symptoms and signs. Both were treated with hyperbaric oxygen therapy in addition to standard resuscitative techniques with rapid improvement. Sodium nitrite and hyperbaric oxygen therapy are suggested specific treatments. The literature and the two case reports support the use of hyperbaric oxygen therapy in the treatment of acute hydrogen sulfide toxicity. Hyperbaric oxygen therapy has also been reported to have a role in the prevention of both short- and long-term neurological toxicity. PMID- 11482867 TI - Another cause for bilateral haemorrhagic otorrhoea following trauma. AB - Bleeding from the external auditory meatus (haemorrhagic otorrhoea) following head trauma is usually attributed to a basal skull fracture. However, it may also result from unsuspected facial trauma. We describe a case that highlights an usual but important differential diagnosis for bilateral haemorrhagic otorrhoea. PMID- 11482868 TI - Prevertebral haematoma after thrombolysis: an unusual cause of airway obstruction. AB - A case report of prevertebral haematoma causing airway obstruction following thrombolytic therapy for acute myocardial infarction is presented. The management options for airway obstruction, after thrombolytic therapy, are discussed. PMID- 11482869 TI - Poisoning with organophosphorus compounds. PMID- 11482870 TI - Absence of ice-cream interference with the adsorption of paracetamol onto activated charcoal: comment. PMID- 11482871 TI - Stem cell plasticity: molding the future of tissue development and repair. PMID- 11482872 TI - Hepatic "stem" cells: coming full circle. AB - Activation, proliferation, and differentiation of a distinct phenotype of cells, called oval cells, are observed after severe hepatic injuries in which the proliferation of existing hepatocytes is inhibited. Under those conditions, oval cells can act as bipotential progenitors of the two types of epithelial cells within the liver, hepatocytes and bile ductular cells. Oval cells are also believed to play a role in the hepatocellular carcinoma and cholangiocarcinoma development; although circumstantial data are available, no direct evidence exists to support this theory. Oval cells have usually been thought to be the progeny of an hepatic stem cell, native to the liver. Recently, however, we, as well as others, have obtained clear evidence that in the rodents, hepatic oval cells, or at least a fraction of them, can derive from a precursor cell of bone marrow origin. The rodent data have been supported by recent findings that human bone marrow cells are capable of becoming hepatocytes and cholangiocytes as well. Having shown that oval cells can be derived from an extrahepatic source, we now have the technology to address many unanswered questions in oval cell origin, fate, and physiology through the use of sex-mismatched bone marrow transplants. PMID- 11482873 TI - Human mesenchymal stem cells persist, demonstrate site-specific multipotential differentiation, and are present in sites of wound healing and tissue regeneration after transplantation into fetal sheep. AB - Prenatal transplantation of stem cells is an exciting frontier for the treatment of many congenital diseases. The fetus may be an ideal recipient for stem cells, as it is immunologically immature and has rapidly proliferating cellular compartments that may support the engraftment of transplanted cells. Mesenchymal stem cells (MSC), given their ability to differentiate into multiple cell types, could potentially be used to treat diseases such as osteogenesis imperfecta, muscular dystrophy, and other mesenchymal disorders that can be diagnosed in utero. We have shown, using a human-sheep in utero xenotransplantation model, that human MSC have the ability to engraft, undergo site-specific differentiation into multiple cell types, and survive for more than 1 year in fetal lamb recipients. In addition, in this model MSC-derived cells appear to be present in increased numbers in wounded or regenerating tissues. This observation warrants further studies of the biology of MSCs following systemic or site-directed transplantation. PMID- 11482874 TI - Hematopoietic progenitors and stem cells in murine muscle. AB - We investigated hematopoietic capabilities of murine skeletal muscle using methylcellulose culture and transplantation into lethally irradiated mice. Muscle mononuclear cells (MNC) contained colony-forming cells and long-term engrafting cells. Studies using chimeric mice indicated a bone marrow origin of the hematopoietic cells in the muscle. We then separated muscle MNC by FACS sorting into Ly-5-positive cells and Ly-5-negative cells and analyzed their hematopoietic capability in vitro and in vivo. The hematopoietic progenitors and stem cells were present only in the Ly-5-positive fraction. PMID- 11482875 TI - Flavopiridol induces apoptosis and caspase-3 activation of a newly characterized Burkitt's lymphoma cell line containing mutant p53 genes. AB - Burkitt's lymphoma cell lines have been important in vitro models for studying the pathogenesis of Burkitt's lymphoma (BL) and for exploring new treatment strategies. A new EBV(-) Burkitt's lymphoma cell line (GA-10) was established from a patient with a clinically aggressive, chemorefractory BL and characterized. Although functional p-glycoprotein could not be demonstrated by dye-efflux assays, both p53 genes were mutated in the GA-10 cells, perhaps contributing to the resistant phenotype of the original neoplasm. Two properties of BL cells which may be useful targets for novel cytotoxic therapeutics are their surface expression of CD77, the receptor for Shiga toxin (Stx), and their high rate of proliferation. Expression of CD77 on the GA-10 cells was heterogeneous in that certain subclones expressed high levels of CD77 and correspondingly exhibited strong growth inhibition by Stx while others showed low levels of CD77 expression and weak Stx-induced growth inhibition. Flavopiridol, a potent inhibitor of cell cycle progression through G1 and G2, induced cytotoxicity of the GA-10 cells with an LC(50) of approximately 40 nM vs 70 nM for HL-60 cells (P < 0.05). The concentrations of flavopiridol at which only 10% of the cells were viable (LC(10)) were approximately 280 nM for the GA-10 cells and 520 nM for the HL-60 cells (P < 0.05). Dose-related induction of apoptosis in response to flavopiridol was demonstrated in the GA-10 cells by morphology, TUNEL assay, and activation of caspase-3. Flavopiridol was also cytotoxic to seven other BL cell lines tested. These data suggest that flavopiridol may have therapeutic value in the treatment of Burkitt's lymphoma. PMID- 11482876 TI - Suggestions for a new paradigm of cell differentiative potential. PMID- 11482877 TI - Adult rat and human bone marrow stromal stem cells differentiate into neurons. PMID- 11482878 TI - 5-Azacytidine modulates the response of sensitive and multidrug-resistant K562 leukemic cells to cytostatic drugs. AB - In an endeavor to improve responsiveness of tumor cells to drug combination treatments, we analyzed the effect of 5-azacytidine (5AC) as a model compound for a new class of drugs, DNA-demethylating agents. We used parental K562/WT chronic myelogenous leukemia cells and a multidrug-resistant subline thereof, K562/ADM. Multidrug-resistant cells were more resistant to daunorubicin, but more sensitive to cisplatin than parental K562 cells as measured by growth inhibition and apoptosis assays. Resistance to daunorubicin can be explained by amplification of the MDR1 drug transporter gene. Cisplatin induced more DNA damage in specific genes and in the entire genome of K562/ADM cells compared to K562/WT cells using PCR stop assays and atomic absorption spectroscopy. Pretreatment with 5AC modulated the response of K562/ADM cells toward MDR-type drugs (daunorubicin, vincristine, etoposide) and reduced function and expression of MDR1 as analyzed by flow cytometry and RT-PCR. Analysis of CpG island methylation in the promotor region of the MDR1 gene by bisulfite sequencing and a methylation-sensitive HpaII digestion/PCR approach revealed that methylation of the MDR1 promotor of K562/ADM cells was greater than in K562/WT cells. 5AC treatment completely abolished MDR1 promotor methylation. The unexpected observation that DNA demethylation by 5AC rather decreases than increases MDR1 expression in K5612/ADM cells points to still unexplored sequences in the MDR1 promotor whose transcriptional activity may be affected by the methylation status. 5AC pretreatment also modulated K562/WT and K562/ADM cells to non-MDR-type drugs such as cisplatin and increased cisplatin-induced DNA damage. PMID- 11482879 TI - Differentiation versus maturation of neoplastic hematopoietic cells: an important distinction. PMID- 11482880 TI - Iron status and HFE genotype in erythrocyte pyruvate kinase deficiency: study of Italian cases. AB - We evaluated the iron status and searched for mutations C282Y and H63D in the hereditary hemochromatosis gene (HFE) in 34 pyruvate kinase (PK)-deficient patients from 29 unrelated families. Nine had received multiple transfusions. Thirteen of the 25 nontransfused patients displayed increased serum ferritin concentration, in the absence of conditions known to raise this parameter. HFE genotype was abnormal in 9 of 34 patients. The allele frequency was 1.8% for mutation 845G--> (C282Y) and 16.1% for mutation 187C-->G (H63D). Nontransfused subjects with abnormal genotype had serum ferritin and transferrin saturation values significantly higher than those with wild-type genotype. Of the 12 adult nontransfused patients with increased iron status parameters, 1 was C282Y homozygous, 1 compound heterozygous for C282Y and H63D, 3 H63D heterozygous, and 7 had a normal HFE genotype. Serum ferritin and transferrin saturation were not related to hemoglobin, reticulocytes, and bilirubin concentration. At multivariate analysis serum ferritin was independently associated with age and gender, but not with splenectomy and HFE genotypes. The retrospective evaluation of the iron status profile of 10 patients (3 with abnormal and 7 with wild-type HFE genotype) with at least 10 years follow-up showed that overt iron accumulation requiring iron chelation had occurred only in the 3 patients (2 of whom were splenectomized) with the mutated HFE gene. PMID- 11482881 TI - Second malignant neoplasms after successful treatment of childhood cancers. AB - Improved treatment and supportive care have increased the survival of children diagnosed with cancer. This success has resulted in a growing population at risk of long-term complications of therapy, including secondary malignancy. These neoplasms may result from the direct effect of the modalities used in treatment of the primary tumor, more indirect effects of the treatment or supportive care, the genetic predisposition of the patient, or to interactions among these factors. The increasing success of cancer therapy is producing a rapidly growing population of patients at risk of second malignancy. This is a result of the increasing intensity of treatments and the increasing duration of survival, which provides the time to manifest the late effects of therapy. The concept that a patient is "cured" at some arbitrary time after treatment does not diminish the need for follow-up of all cancer survivors to identify and treat secondary malignancies. These risks have led to an increased effort to define phenotypic and genotypic categories of patients that may be cured with less intensive therapy and to develop molecularly targeted drugs that have fewer noxious effects on normal tissues. PMID- 11482882 TI - Cloning and characterization of Hepp, a novel gene expressed preferentially in hematopoietic progenitors and mature blood cells. AB - Through differential screening of mouse hematopoietic stem cell (HSC) and progenitor-subtracted cDNA libraries we have identified a progenitor cell specific transcript that represents a novel gene, named Hepp (hematopoietic progenitor protein). The mouse Hepp gene encodes a protein of 237 amino acids with no detectable known functional domains or motifs. Lack of invertebrate orthologs and a high degree of evolutionary conservation of the peptide sequence in vertebrate species (zebrafish, mouse, human) suggest that the Hepp gene could have conserved although as yet unknown function in vertebrates. Mouse Hepp shows a restricted expression pattern in adult tissues and is transcribed at a very low level in heart, lung, spleen, and thymus and at a higher level in muscle. During embryonic hematopoiesis Hepp is not expressed in mouse fetal liver HSC (Sca-1(+)c kit(+)AA4.1(+)Lin(-) cells), but is abundantly transcribed in the population of hematopoietic progenitors (AA4.1(-) cells). Similarly, during adult hematopoiesis Hepp is not transcribed in the highly enriched population of bone marrow HSC (Rh 123(low)Sca-1(+)c-kit(+)Lin(-) cells), but its expression is upregulated as a greater heterogeneous population of bone marrow HSC (Lin(-)Sca-1(+) cells) differentiates into progenitors (Lin(-)Sca-1(-) cells) and more mature lymphoid and myeloid cell types. A restricted pattern of expression in adult tissues and preferential expression in both fetal and adult hematopoietic progenitors and mature blood cells suggest that Hepp could be involved in molecular regulation of HSC and progenitor cell lineage commitment and differentiation. PMID- 11482883 TI - Osteogenesis and bone-marrow-derived cells. AB - This paper addresses some of the important aspects of stem cell commitment to the bone cell lineage examining the various types of precursor cells, their responses to cytokines and other extracellular influences, and recent observations on the biochemical and molecular control of lineage-specific gene expression. The process of osteopoiesis involves the proliferation and maturation of primitive precursor cells into functional osteoblasts. The bone cells purportedly originate from mesenchymal stem cells that commit to the osteogenic cell lineage becoming osteoprogenitor cells, preosteoblasts, osteoblasts, and osteocytes. Further understanding of this developmental process requires that lineage-specific markers be identified for the various populations of bone cells and their precursors, that cell separation techniques be established so that cells of the osteogenic lineage can be purified at different stages of differentiation, and that these isolated cells are studied under serum-free, chemically defined conditions. PMID- 11482884 TI - Rapid identification of hemoglobin variants by electrospray ionization mass spectrometry. AB - The precise identification of human hemoglobin variants, over 700 human hemoglobin variants are known, is essential for prediction of their clinical and genetic significance. A systematic approach to their rapid identification is described. Traditionally this requires protein or DNA characterization which entails lengthy analytical procedures. To overcome these obstacles a rapid approach to variant hemoglobin identification has been developed using conventional phenotypic methods combined with electrospray ionization-mass spectrometry (ESI-MS). The latter requires only a small amount of whole blood (10 microl) but in most cases 2 microl would have been sufficient and no preanalytical steps, such as separation of red cells or globin chains, are necessary. Aged, hemolyzed blood samples can also be analyzed. This approach has been used to positively identify 95% of the variants in over 250 samples. The remaining 5% in which a variant was detected by phenotypic techniques were not resolved by mass spectrometry. Ninety-nine different abnormalities comprising 36 alpha-chain variants, 59 beta-chain variants (including 2 extensions), and 4 hybrid hemoglobins were identified. These include 15 novel variants. The application of ESI-MS described requires approximately 1 h to prepare and analyze each sample and has minimal reagent costs. The turnaround time on a single sample can be as little as 2 h. This technique can now be considered a useful additional tool for reference laboratories. PMID- 11482885 TI - Neuronal differentiation of mouse embryonic stem cells: lineage selection and forced differentiation paradigms. AB - Primitive embryonic stem cells are an ideal starting cell population for studies of gene expression and lineage segregation during development. Despite their potential, it has been difficult to determine culture conditions that cause single-lineage differentiation of these pluripotent cells. Both genetic and epigenetic approaches have been taken to promote neuronal differentiation of embryonic stem cells, including aggregation, exposure to the nonspecific teratogen/morphogen retinoic acid, low-density culture, exposure to growth/differentiation factors, and forced differentiation following expression of lineage-restricted "developmental control" genes. In the current investigation, a hybrid approach involving genetic techniques of "lineage selection" or "forced differentiation" has been employed to develop primitive neural progenitor cell lines. These lines form an important starting point to examine the cascades of gene expression (and inhibition) during neuronal and glial lineage segregation, to study growth factor effects on neural differentiation, and ultimately to provide a source of cells for transplantation to a damaged nervous system. PMID- 11482886 TI - Evaluation of a tetrazolium salt test to determine absence of live mycobacteria in tuberculin purified protein derivative. AB - Current methodology to determine absence of live mycobacteria in tuberculin purified protein derivative (PPD) takes up to 8 weeks to perform and may also involve testing on animals. In this paper we describe an in vitro test utilising the tetrazolium salt, 2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5 carboxanilide (XTT) to monitor the absence of live Mycobacterium tuberculosis (Mtb) in PPD. In the presence of live cells XTT is converted to a coloured formazan product that can be measured spectrophotometrically. Live mycobacteria present in spiked PPD were detected by a marked change in optical density above background levels. This test is easy to perform and is complete in just 48 hr. PMID- 11482887 TI - Pasteurella multocida toxin type D serological assay as an alternative to the toxin neutralisation lethality test in mice. AB - The objective of this work was to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against Pasteurella multocida toxin type D, that correlated to a mouse lethality test. Currently, the mouse lethality test is one of several tests used world-wide to evaluate serological responses in animals immunised with vaccines containing toxoids. The mouse lethality test involves injecting mice with a mixture of toxin and test serum sample (from animals that have been vaccinated with a toxoid), and then determining antibody titre of the test serum from the number of mice that survive. Thus, the titre calculated is based on the neutralising activity of the test serum. The mouse lethality test requires large numbers of animals and causes severe distress to the animals. Organisations world-wide are working towards alternatives to animals in the development and control of biological products for human and veterinary use. Additionally, the mouse lethality test is labour-intensive, costly and lacks robustness and may be difficult to reproduce between different technicians. We have developed a double sandwich ELISA to measure anti- P. multocida toxoid type D antibodies in swine serum. Sera from swine immunised with vaccines containing type D toxoid showed good correlation to the mouse lethality assay (Spearman analysis=0.94 and Pearson analysis=0.84). When compared to the mouse lethality test, titres obtained using the ELISA format had higher correlation with protective immunity (i.e., lower turbinate atrophy) following challenge with virulent P. multocida. The ELISA assay is more robust, reproducible and costs less than the mouse lethality assay; and it complements efforts to reduce the use of animals in testing. PMID- 11482888 TI - Induction of group 17 specific antibodies by pneumococcal type 17F and 17A polysaccharide vaccines. AB - Pneumococcal capsular polysaccharide group 17 contains two distinct serotypes, 17F and 17A. Pneumococcal group 17 is present in the licensed 23 valent polysaccharide vaccines. One such vaccine contains type 17A, while the other vaccine contains type 17F. The purpose of these studies was to determine the extent of cross-protection that could be expected, as both type 17F and 17A cause disease. The antibody responses of one group of adults to a vaccine containing type 17F was compared to that of another group that received a type 17A containing vaccine. By ELISA the 17A vaccine induced more cross-reactive antibodies. Opsonophagocytic antibodies are a good predictor of protection and both vaccines induced antibodies opsonic for both 17F and 17A. We conclude that either 17F or 17A will provide similar protection against group 17 disease. PMID- 11482889 TI - Scrapie removal using Planova virus removal filters. AB - As a possible method for reducing the risk of transmissible spongiform encephalopathy (TSE) infection, Planova virus removal filters were tested for their ability to remove scrapie agent ME7. Albumin solution was spiked with high titre ME7 and filtered through three different pore sizes of Planova filters. Infectivity of the pre- and post-filtration samples was assayed in log dilutions by intracerebral inoculation into C57B1/6 mice. Filtration of albumin solution in the absence or presence of a detergent (Sarkosyl) with Planova 35N (35+/-2 nm mean pore size) removed the contaminating scrapie agent with reduction factors of 4.93 log10 and 1.61 log10, respectively. Filtration, both in the absence and presence of detergent with Planova 15N (15+/-2 nm mean pore size), and in the presence of detergent with Planova 10N (9+/-2 nm mean pore size), showed high levels of scrapie reduction of >5.87 log10, >4.21 log10, and >3.80 log10, respectively, with no residual infectively detected in any of the filtrate samples. The effectiveness of Planova 35N filtration for the removal of infectivity of this TSE agent is greatly reduced in the presence of a strong detergent, but Planova filters with 15 nm or smaller pore size membranes can remove such infectivity at high reduction rates. PMID- 11482890 TI - Development of a pharmaceutical apotransferrin product for iron binding therapy. AB - High-dose chemotherapy of patients with haematological malignancies results in extracellular iron accumulation and appearance of non-transferrin-bound iron, which is thought to predispose the patients to septic infections and contribute to organ toxicity. We describe the development of a human plasma-derived apotransferrin product for iron binding therapy. The product is purified from Cohn fraction IV of human plasma by two ion exchange chromatography steps and ultrafiltration. The process comprises solvent detergent treatment as the main virus inactivation step and 15 nm virus filtration and polyethylene glycol precipitation as removal steps for physico-chemically resistant infectious agents. Product characterization by electrospray and MALDI-TOF mass spectrometry indicated no other chemical modifications than N-linked glycan chains and disulphide bonds, except minor oxidation. The purity of the product was more than 98%, main impurities being IgG, IgA and hemopexin. The product had intact iron binding capacity and native conformation. A stable liquid formulation for the finished product was developed. The product has proved safe and well tolerated in early clinical trials in iron binding therapy. PMID- 11482891 TI - Development of control serum for microbial antibody tests. AB - Control sera are an essential component of in-vitro clinical diagnostic reagents. We have developed a control serum for HBsAg, anti-HCV, anti-HIV and anti- Treponema pallidum testing. Since this control serum is prepared from human plasma, we paid special attention to viral safety. We incorporated three viral inactivation methods into the manufacturing process that maintain the necessary characteristics of a stable control serum. PMID- 11482892 TI - Detection of hepatitis C virus RNA in alpha interferon derived from in vitro culture of leukocytes of human origin. AB - In order to assure the virological safety of blood products, in addition to serological testing of individual donations and virus inactivation steps undertaken during manufacture, routine PCR testing for HCV RNA of starting materials (plasma, cells), intermediates or final product is necessary. The aim of this study was to determine the rate of HCV RNA positive batches of human native leukocyte interferon during large-scale production. Our findings indicate the presence of HCV RNA in 6.1% batches despite acidification of intermediates in order to inactivate Sendai virus. PMID- 11482893 TI - Interfering effect of diphtheria-tetanus-acellular pertussis combined (DTaP) vaccines on the bacterial endotoxin test. AB - We evaluated applicability of the endotoxin test to DTaP vaccines. Although no DTaP vaccine batches showed immediate interference with the activating activity of spiked endotoxin on the clotting of Limulus amaebocyte lysate (LAL), a gradually progressing interference depending on the time after spiking was seen for some of the batches. The interfering DTaP vaccine batches, however, showed no significant effect onin vitro TNF-alpha induction in RAW264.7 cells and pyrogenicity in rabbits of spiked endotoxin. Aluminium hydroxide gel in the vaccines was suggested to be one of the causes of the interference. Accordingly, a careful evaluation of the interfering effect was assumed crucial for the effective application of the endotoxin test to check residual biological activity of endotoxin in DTaP vaccines. PMID- 11482894 TI - Characterization and regulation of constitutive transport intermediates involved in trafficking from the trans-Golgi network. AB - Transport vesicles or containers (TCs) mediate constitutive protein transport between the trans-Golgi network (TGN) and the plasma membrane. A key question is the nature and regulation of these transport containers or intermediates. We have used a trans-Golgi network resident, TGN38, to investigate TC formation. TGN38 is a recycling membrane glycoprotein that moves to the cell surface via constitutive membrane traffic and returns via the endosomal pathway. An in vitro assay to measure TC formation was devised using rat liver Golgi membranes, cytosolic factors and ATP. Transport intermediates containing TGN38 were produced and found to be smooth vesicles and tubules of up to 200 nm in length. These membrane enclosed structures contain different constitutively secreted membrane glycoproteins, including molecules involved in immune functions such as MHC Class I and the polymeric Ig receptor, showing that these intermediates correspond to TCs that have been previously identified in vivo. Importantly, TC formation can be stimulated or inhibited by protein kinase and phosphatase inhibitors, showing regulation by intracellular signalling pathways. PMID- 11482895 TI - Contaminants within bacterial plasmid preparations trigger apoptosis in liposome transfected OVCAR3, but not in SKOV3 or AZ224 human ovarian cancer cells. AB - Toxicity associated with plasmid/liposome transfection of eucaryote cells has been attributed to the inherent toxicity of cationic lipid formulations and also to bacterial contaminants of plasmid DNA preparations, such as lipopolysaccharides (LPS). Certain plasmid preparations were observed to trigger apoptosis in DNA/liposome transfected OVCAR3 human epithelial ovarian cancer cells. In contrast, AZ224 and SKOV3 cells were unaffected under the same conditions. Agarose gel electrophoresis with recovery of the plasmid DNA removed the toxic component, but not purification by phenol/chloroform extraction or isopicnic CsCl ultracentrifugation. The toxicity of individual preparations correlated with the concentration of bacterial LPS. However, polymixin B could not neutralise the toxicity and neither could the effect be reproduced by the addition of bacterial LPS to non-toxic plasmid preparations. Surprisingly, the conditioned medium of OVCAR3 cells undergoing apoptosis was found to kill non transfected OVCAR3 cells but not AZ224 or SKOV3 cells. This observation illustrates the possibility that unpredictable contaminants of bacterial plasmid preparations are able to cause cell death in the context of plasmid/liposome transfection in a cell-type specific way. It emphasizes the importance of achieving maximal plasmid DNA purity when performing DNA transfection experiments that focus on cell survival. PMID- 11482896 TI - Expression of BAX in cell nucleus after experimentally induced apoptosis revealed by immunogold and embedment-free electron microscopy. AB - The unique combination of immunocytochemistry with embedment-free electron microscopy was applied for precise and specific localisation of BAX in the human colon adenocarcinoma COLO 205 cell line stimulated to undergo apoptosis by camptothecin (DNA topoisomerase I inhibitor). Camptothecin-induced apoptosis was associated with redistribution of BAX from cytosol to organelle membranes: mitochondria, Golgi apparatus, endoplasmic reticulum and via nuclear envelope pores to the nucleus, occurring within 60-180 min of cell exposure to the drug. An increase in BAX immunoreactivity on fine filaments and the lamina-pore complex of the nuclear matrix was also observed. The increase in BAX expression in the nuclear area of camptothecin-treated COLO 205 cells was confirmed by quantitative analysis using laser scanning cytometry. The subcellular translocations of BAX preceded the appearance of any morphological symptoms of apoptosis. PMID- 11482897 TI - Effect of hyaluronan on MMP expression and activation. AB - Matrix metalloproteases (MMPs) play a crucial role in tissue remodelling in a variety of physiological and pathological processes. Hyaluronan is also involved in the same processes. Several cytokines and growth factors are involved in the regulation of the biosynthesis of hyaluronan and also of MMPs. The activity of MMPs has been shown to be regulated at the level of transcription and activation of the zymogen form. In order to explore the possible relationship between matrix components and especially hyaluronan, we studied the effect of hyaluronan on MMP expression (biosynthesis and activation) in the culture of human skin fibroblasts and corneal keratocytes (explant cultures and cell cultures). These cells were shown to exhibit distinct phenotypes as far as matrix biosynthesis is concerned. Using a synthetic substrate N-Suc(ala)3pNA, we measured elastase-type endopeptidase activity produced by fibroblasts and keratocytes and characterized the MMPs by zymography. Hyaluronan added to fibroblast cultures stimulated the membrane-bound elastase-type endopeptidase activity in a dose dependent fashion. Similar results were obtained with keratocyte cultures. In the presence of 1 mg/ml hyaluronan there was an increase in MMP expression and also an activation of latent MMPs both by fibroblasts and keratocytes. PMID- 11482898 TI - Differentiation potential of intestinal mesenchyme and its interaction with epithelial cells: a study using beta-galactosidase-expressing fibroblast lines. AB - Rat intestinal fibroblast lines (F1:G9 and A1:F1) differing in their potential to support intestinal mucosal development were marked with reporter genes to investigate their differentiation potential. The fibroblasts were transfected with plasmids expressing either beta-galactosidase (with or without a nuclear localisation signal) or green fluorescent protein (GFP). Transfection using Tfx50 or Fugene was more efficient than electroporation. The expression of beta galactosidase was more stable and stronger than GFP. Cells were optimally labelled using the plasmid pL27B-GAL, and sub-clones with a strong and uniform nuclear expression of beta-galactosidase were isolated. These clones expressed beta-galactosidase even after prolonged passage in the absence of selection. The beta-galactosidase tagged lines (F1:G9gal and A1:F1gal) retained the morphological characteristics, viability and differentiation properties of the parental non-transfected lines. In co-culture with a colorectal tumour cell line Caco-2, the F1:G9gal and A1:F1gal cells differed in their morphological organisation but this did not change their expression of smooth muscle alpha actin. PMID- 11482899 TI - Integrin and talin in the jellyfish Podocoryne carnea. AB - We have isolated an integrin-beta and -alpha subunit from Podocoryne carnea (Cnidaria, Hydrozoa) and studied their expression in the life-cycle and during cell migration, in vitro transdifferentiation and regeneration. Comparison of the integrin expression pattern with a Podocoryne talin homologue by RT-PCR demonstrates that all three genes are maternal messages and continuously expressed in the life-cycle, in medusa development and in all medusae tissues. In situ hybridisation experiments confirm co-expression of both integrin subunits in the different life-stages. Integrin expression was furthermore studied in isolated striated muscle induced to transdifferentiate to new cell types, or grafted on ECM where the muscle adheres and migrates. Integrin expression was maintained continuously throughout both processes. These results suggest that in Podocoryne carnea processes such as cell migration and differentiation are not controlled by up- or downregulation of alternative integrin subunits, but by a single integrin heterodimer which activates different downstream signalling cascades. PMID- 11482900 TI - Erythrocyte sodium/hydrogen exchange inhibition by (-) epicatechin. AB - Epicatechin, a flavonoid belonging to the group of compounds collectively called catechins, have been reported to possess insulin-like properties. Besides their anti-diabetic properties, catechins also show growth inhibition. Since cytosolic pH (pHi) plays a role in cell proliferation and the Na/H exchanger (NHE) is the major pH (pHi) regulatory mechanism, we undertook in vitro studies with human erythrocytes to examine the effect of (-) epicatechin (EC) on the NHE1 isoform. NHE activity was measured in eight healthy volunteers, eight type 1 diabetics, and nine type 2 diabetics, following 30 min incubations at 37 degrees C with either 1 mM epicatechin, 10(-9) M insulin or solvent alone. NHE activity was elevated in both groups of patients (P< 0.05). Epicatechin caused a 93% decrease in Na/H antiport activity in health controls, 89 and 86% in type 1 and type 2 diabetics, respectively (P< 0.001). Insulin caused a 36% decrease in antiport activity only in the type 2 diabetic group (P< 0.05). The strong inhibition of erythrocyte NHE1 (the ubiquitously present isoform) by epicatechin may have important implications. NHE1 inhibition could be one of the major mechanisms underlying the antiproliferative effects of catechins. PMID- 11482901 TI - Linoleic and linolelaidic acids differentially influence proliferation and apoptosis of MOLT-4 leukaemia cells. AB - The effects of varying concentrations of linoleic acid and its transisomer linolelaidic acid on the proliferation the ultrastructural morphology of MOLT-4 T lymphoblastic leukaemia cells were investigated. At 2 and 4 days after exposure to the fatty acids, the cells were counted by flow cytometry and observed by electron microscopy. After 4 days of treatment, linoleic acid was growth stimulatory at concentrations of 200 microM or less, but was markedly inhibitory at 400 microM. In contrast, linolelaidic acid stimulated proliferation at concentrations of 100 and 200 microM, but inhibited cell growth at 400 microM. Cells treated with 400 microM linoleic acid displayed dense accumulations of characteristic lipid globules and glycogen granules, and exhibited ultrastructural evidence of apoptosis including vacuolization, membrane blebbing and chromatin margination at the nuclear periphery. These results support the notion that geometrical isomerism and concentration of polyunsaturated fatty acids influence the proliferative destiny of cancer cells. Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed a previously documented larger alternatively spliced p53 gene transcript in MOLT-4 cells cultured under reduced serum conditions. However, only wild-type p53 transcripts were amplified by RT-PCR of MOLT-4 cells exposed to phytohaemagglutinin, linoleic acid or linolelaidic acid. PMID- 11482902 TI - Estrogen downregulates the number of caveolae and the level of caveolin in uterine smooth muscle. AB - Estrogen and progesterone, while regulating uterine functions, also regulate the number of caveolae and the level of caveolin. Large numbers of caveolae, as well as elevated expression of caveolin-1 and caveolin-2 isoforms in the myometrium of ovariectomised (OVX) rats were detected. 17beta-estradiol (E2) has a downregulating effect: the treatment of OVX rats with E2 (5 microg/animal) reduced the formation of caveolae by approx. 90%. Western blots clearly demonstrated the reduction of membrane caveolin-1 and -2 content. Progesterone treatment (2.5 mg/animal) alone did not cause any substantial change, but prevented the effect of estrogen. Control experiments showed that the quantity of Na+/K+-ATPase, a plasma membrane protein excluded from caveolae, was not downregulated by E2. The administration of the pure estrogen receptor (ERalpha) antagonist ICI 182,780 (1 mg/animal) not only compensated for the inhibitory effect of E2, but further increased the level of caveolin-1 in the myometrium of OVX rats and facilitated the formation of caveolae by approximately 70%. In contrast, the partial antagonist tamoxifen (1 mg/animal) mimicked the effect of estrogen. The amount of caveolin also changed during pregnancy. During the first half of pregnancy the expression of caveolin was suppressed, but it gradually increased until delivery. Our results indicate that the formation and number of caveolae are influenced by the physiological state of the uterus in a hormone dependent manner. PMID- 11482903 TI - Morphological alteration of peritoneal mast cells and macrophages in the mouse peritoneal cavity during the early phases of an allergic inflammatory reaction. AB - We investigated the presence of mast cell granules in macrophages following an in vivo model of an allergic reaction. Injection of ovalbumin (100 microg) into the peritoneal cavity of sensitised mice produced a rapid (within 2 h) influx of neutrophils followed by a slower (after >4 h) eosinophil migration. Ovalbumin treatment induced a high incidence (approximately 50%) of mast cell degranulation compared to control phosphated-buffered saline-treated mice. The majority (approximately 90%) of peritoneal macrophages contained mast cell granules as early as 2 h post-ovalbumin, with lower values at later time-points, as determined by staining with Toluidine blue and Berberine sulphate. This was confirmed by electron microscopy which enabled us to identify the complex mast cell granule sub-structural components in macrophage phagosomes. In conclusion, we used histochemical and ultrastructural analyses to show that mast cell granules become internalised with macrophages during the early stages of an experimental allergic reaction. PMID- 11482904 TI - Phosphorylation of filamin (ABP-280) regulates the binding to the lipid membrane, integrin, and actin. AB - Actin-binding protein (ABP-280; filamin) is a phosphoprotein present in the periphery of the cytoplasm, where it can cross-link actin filaments, associate with lipid membranes, and bind to membrane surface receptors. Given its function and localization in the cell, the hypothesis that it serves as a substrate for p56lck, a lymphocyte-specific member of the src family of protein tyrosine kinases associated with cell surface glycoproteins is considered. The results suggest conformationally-induced regulation of filamin (ABP-280). PMID- 11482905 TI - Implication of annexin 1 in phagocytosis: effects of n-terminal domain deletions and point mutations of the phosphorylation site Ser-27. AB - Directed mutagenesis, in the form of deletions and point mutations, was used to investigate the regulatory importance of the N-terminal domain of annexin 1. Wild type and mutant forms were fused to green fluorescent protein (GFP) to track their localization and introduced in to J-774A.1 cells by transfection. The fusion of annexin 1 to GFP at the N- or C-terminal end did not alter the cellular distribution or co-localization with phagosomes. The effects of mutations were determined according to these characteristics. The prominent effect resulted from S27E mutation which mimics the phosphorylated state of Ser-27. Although still retaining the granular structures in the cytoplasm, S27E annexin 1 failed to associate with the phagosomal protein complex. This suggests an essential regulatory role of the phosphorylation of residue 27 in annexin 1 function. PMID- 11482906 TI - The effect of cytochalasin J on kinetochore structure in PtK1 cells is mitotic cycle dependent. AB - Mitotic PtK1 cells were arrested in mitosis with nocodazole to determine the effect of cytochalasin J (CJ) on kinetochore structure in arrested and nocodazole released cells. In previous studies it was shown that CJ had a more pronounced effect on alteration of kinetochore structure and spindle microtubule (MT) architecture when applied during prophase or prometaphase. In this study, mitotic cells were treated at preanaphase for 10 min with 1 microg/ml nocodazole, or in 1 microg/ml nocodazole and 10 microg/ml CJ to allow for the advancement of the 'mitotic clock'. Thus it can be determined if either changes in the timing of mitosis, the maturation of the kinetochore, and/or the lack of MT connection to the kinetochore affects the ability of CJ to detach or alter the attachment of chromosomes to the developing spindle. Preanaphase cells treated with 1 microg/ml nocodazole for 10 min and released into 10 microg/ml CJ showed significant changes in MT organization and kinetochore structure. MTs nucleated at the centrosome are fragmented and kinetochore structure was significantly altered showing only two laminae with few MTs inserted into this structure. Preanaphase cells treated with 1 microg/ml nocodazole and 10 microg/ml CJ for 10 min and released into 10 microg/ml CJ showed similar, but more pronounced, effects on kinetochore structure and spindle MT organization. We interpret these results to suggest that CJ treatment has a greater effect on MT attachment and kinetochore structure in nocodazole pre-treated cells, where the kinetochore structure is mature and the mitotic cycle has been advanced. PMID- 11482907 TI - Transcription of Fc(gamma) receptors in different rat liver cells. AB - Expression of Fc(gamma)-receptors (Fc(gamma)Rs) in different liver cells is poorly characterised. In the present study, transcription of Fc(gamma)Rs in different rat liver cells was examined by means of Northern blot analysis and reverse transcriptase-polymerase chain reactions. We found that both Kupffer and liver endothelial cells produce mRNA for Fc(gamma)RIIB2, Fc(gamma)RIII and the gamma-chain, whereas the level of Fc(gamma)RIIB1 mRNA is negligible. In contrast, parenchymal cells produce no Fc(gamma)R mRNA. PMID- 11482908 TI - Effects of insulin-like growth factor-binding protein 2 and an IGF-type I receptor-blocking antibody on apoptosis in human teratocarcinoma cells in vitro. AB - Human teratocarcinoma cells (Tera-2) deprived of serum undergo programmed cell death which can be counteracted by simultaneous addition of IGF-I or IGF-II. This protective effect of IGFs was specific in the sense that both addition of IGF binding protein-2, and blocking of the IGF-type I receptor by a specific antibody, both resulted in an increased apoptotic rate. PMID- 11482909 TI - Ultradian rhythms and the lux-gene reporter system. AB - 'Noisy' bioluminescence of an unstirred and exposed-to-air microculture of lux gene fused Escherichia coli was interpreted, assuming two oscillatory processes: cyclic induction of the lac promoter, resulting in oscillatory reporter lux -gene expression, and oscillatory respiration in the culture. Biological support for the theoretical argument, that two rhythms can interact in cells to produce additional rhythms, is provided. These results, along with previously obtained data on oscillatory bioluminescence, indicate the deterministic nature of observed noise-like fluctuations and promising perspectives of lux gene reporter systems in investigations of ultradian rhythm. PMID- 11482910 TI - gp130-specific antisense oligonucleotides inhibit IL-6 signal inducing junB mRNA transcription in the human hepatoma cell line, HepG2. AB - The biosynthesis of interleukin-6 receptor (IL-6R) and gp130 in vitro was blocked using specific antisense oligonucleotides (ASO) in HepG2 liver cells and the efficacy of various ASOs was tested on the generation of IL-6-induced junB mRNA. We used three ASOs specific for the IL-6 receptor, three specific for gp130 and a control (nonsense) oligonucleotide specific for epsilon-chain of IgE (not expressing in HepG2 cells). Our data indicate that a gp130-specific ASO, g2, was the most effective blocker of IL-6-induced junB mRNA, whilst the IL-6 receptor ASOs alone were ineffective. The mechanism of gene inactivation by ASO treatment was partially elucidated by demonstration of the loss of gp130 mRNA from cells treated with ASOs showing functional efficacy. Our data may help to design antisense oligonucleotides that are effective in therapy (e.g. as anti inflammatory agents) in the future. PMID- 11482911 TI - Release of IL-4 and IL-5 from human peripheral blood mononuclear cells co cultured with Japanese cedar pollen antigen in vitro. AB - The Japanese cedar pollen (JCP) is a major allergen with respect to pollinosis in Japan. It is believed that interleukin-4 (IL-4) and interleukin-5 (IL-5) derived from lymphocytes and other cells play a pivotal role in allergic reactions. We investigated whether the JCP antigen stimulates the release of these cytokines by peripheral blood mononuclear cells (PBMCs). PBMCs from eight adults (five adults with JCP pollinosis and three adults without JCP pollinosis) were co-incubated with purified JCP antigens. IL-4 was released in response to JCP antigens in six of the eight subjects at 24 h and in three subjects at 48 h. IL-4 release at 24 h occurred in all five subjects with JCP pollinosis but in only one of the three subjects without pollinosis. IL-5 was released in response to the JCP antigen in five of the eight subjects at 24 h and 48 h, including four of the five subjects with JCP pollinosis and one of the three subjects without pollinosis. These results suggest that PBMCs were more likely to release IL-4 and IL-5 in the presence of JCP pollinosis. PMID- 11482912 TI - EECP -- new data on possible mechanisms of action. PMID- 11482913 TI - Fascination with myocardial infarction and normal coronary arteries. PMID- 11482914 TI - Economics of coronary stenting and GPIIb/IIIa blockade. PMID- 11482915 TI - Measuring cardiac power output -- the acid test. PMID- 11482916 TI - Sotalol: a fool's deal? PMID- 11482917 TI - Task Force on Sudden Cardiac Death of the European Society of Cardiology. PMID- 11482918 TI - Enhanced external counterpulsation improved myocardial perfusion and coronary flow reserve in patients with chronic stable angina; evaluation by(13)N-ammonia positron emission tomography. AB - AIMS: The mechanism by which enhanced external counterpulsation therapy exerts its beneficial effects on chronic and symptomatic stable angina is largely unknown. To clarify the mechanism of action of enhanced external counterpulsation, we used(13)N-ammonia positron emission tomography to evaluate myocardial perfusion. METHODS AND RESULTS: This was not a randomized controlled study. Eleven patients (eight male, age: 61.6+/-9.7) with angina pectoris underwent enhanced external counterpulsation therapy for 35 1 h sessions. They underwent a treadmill exercise test and(13)N-ammonia positron emission tomography, both at rest and with dipyridamole, before and after enhanced external counterpulsation therapy. Neurohumoral factors and nitric oxide were also evaluated. Myocardial perfusion increased at rest after therapy (0.69+/-0.27 to 0.85+/-0.47 ml x min(-1) x g(-1), P<0.05). In ischaemic regions, particularly the anterior region, myocardial perfusion at rest and with dipyridamole and coronary flow reserve improved significantly after therapy (at rest: 0.71+/-0.26 to 0.86+/-0.31;P<0.05, with dipyridamole: 1.26+/-0.65 to 1.84+/-0.94;P<0.02, coronary flow reserve: 1.75+/-0.24 to 2.08+/-0.28;P<0.04). Exercise time was prolonged and the time to 1-mm ST depression improved markedly (P<0.01). After therapy, nitric oxide levels increased (P<0.02) and neurohumoral factors decreased. CONCLUSIONS: Enhanced external counterpulsation therapy improved myocardial perfusion at rest and with dipyridamole and was associated with an increased exercise tolerance with(13)N-ammonia positron emission tomography and increased nitric oxide levels. These results suggest that one of the enhanced external counterpulsation mechanisms is development and recruitment of collateral vessels. PMID- 11482919 TI - Clinical characteristics, aetiological factors and long-term prognosis of myocardial infarction with an absolutely normal coronary angiogram; a 3-year follow-up study of 91 patients. AB - OBJECTIVES: The purpose of this study was to evaluate the clinical outcome of a large cohort of patients who suffered an acute myocardial infarction with absolutely normal epicardial coronary arteries at the post-myocardial infarction coronary angiogram. The aetiological and prognostic factors in this population were also analysed. BACKGROUND: Few data exist concerning the outcome, and aetiological and prognostic factors, of patients with myocardial infarction and angiographically absolutely normal coronary arteries. METHODS: Ninety-one patients (34 females/57 males; mean age 50+/-13 years, range 24--78 years) admitted with an acute myocardial infarction had absolutely normal coronary arteries at the angiogram performed 6.2+/-4 days (range 1--15 days) after the myocardial infarction, defined by smooth contours and no focal reduction (NC). Of the 91 NC patients, 71 were evaluated prospectively, alongside a systematic search of all aetiological factors reported in the literature. The NC patients were matched for age, sex, and the same period of myocardial infarction onset with a group of 91 patients with coronary artery stenosis (>50% diameter stenosis) at the angiogram performed 7.3+/-4 days (range 1--15 days) after the myocardial infarction (SC). RESULTS: The percent of smokers was similar between the two groups; higher prevalence rates of coronary heart disease family history, obesity, hypertension, hypercholesterolaemia and diabetes mellitus were found in SC (P=0.043 to 0.0001). In NC, coronary spasm was found in 15.5%, congenital coagulation disorders in 12.8%, collagen tissue disorders in 2.2%, embolization in 2.2%, and oral contraceptive use in 1.1%. Left ventricular ejection fraction at hospital discharge was higher in NC (60%+/-13%) than in SC (55%+/-13%, P=0.04). The mean follow-up was 35 months (range 1--100 months). Kaplan-Meier event-free survival, with the combined end-point defined as death, reinfarction, heart failure and stroke was 75% in NC vs 50% in SC (P<0.0001). Survival rate was 94.5% in NC compared to 92% in SC (ns). Univariate predictors of events in NC were left ventricular ejection fraction (P=0.03), age (P=0.02), diabetes (P=0.01), and smoking (P=0.03). Using Cox multivariate analysis, independent predictors of long-term outcome in NC patients were left ventricular ejection fraction (P=0.003) and diabetes (P=0.004). CONCLUSION: Aetiological factors, predominantly coronary spasm and inherited coagulation disorder, can be detected in only one third of the patients with myocardial infarction and absolutely normal coronary angiograms despite a systematic search in a prospective population. Mortality rates are similar but morbidity is lower in myocardial infarction patients with absolutely normal coronary angiography compared with those with coronary artery stenosis. The only two independent factors predictive of poor outcome in myocardial infarction patients with normal coronary arteries are left ventricular function and diabetes. PMID- 11482920 TI - Microalbuminuria during acute myocardial infarction; a strong predictor for 1 year mortality. AB - AIMS: Urinary albumin excretion increases during acute myocardial infarction but little is known on the prognostic significance and the pathophysiological mechanisms of microalbuminuria in this clinical setting. The primary aim of the study was to examine whether urinary albumin excretion has predictive power for 1 year mortality after acute myocardial infarction. A secondary objective was to gain insight into the pathophysiological mechanisms of increased urinary albumin in myocardial infarction. METHODS AND RESULTS: This is a prospective cohort study conducted in three coronary care units (Northeast Italy). Four hundred and thirty two unselected, consecutively enrolled patients with acute myocardial infarction (66.3+/- 12.3 years of age) were studied. The incidence of mortality was related to the baseline urinary albumin:creatinine ratio. The best cut-off for total mortality approximated to 50 mg x g(-1)on the first day after myocardial infarction, 30 mg x g(-1)on the third day, and to 20 mg x g(-1)on the seventh day. At multivariable Cox analysis, the albumin:creatinine ratio was the strongest among several independent predictors of mortality (adjusted relative risks: 3.6 (95% CI, 2.1--6.2) on the first day, 4.9 (95% CI, 2.9--8.2) on the third day and 4.0 (95% CI, 2.3--6.8) on the seventh day). Independent determinants of urinary albumin were plasma aldosterone on the first day, and inflammatory markers on the third and seventh days. CONCLUSION: Urinary albumin assessed in the first week after acute myocardial infarction is a strong prognostic marker for 1-year mortality. PMID- 11482921 TI - Cost-efficacy in interventional cardiology; results from the EPISTENT study. Evaluation of Platelet IIb/IIIa Inhibitor For Stenting Trial. AB - AIMS: The EPISTENT study has demonstrated that the combined use of abciximab and stenting as an adjunct to PTCA leads to increased event-free survival compared to either using abciximab or stenting alone. However, this combined strategy may be costly and the additional costs have to be weighted against the additional effects. METHOD AND RESULTS: The 6-months efficacy data from the EPISTENT study are combined with Dutch estimates of unit costs. Adding a stent to a procedure with abciximab further decreases the number of revascularizations at an extra cost of Euros 12,000 (95% upper limit (u.l.) Euros 31,000) per additional major adverse cardiac event-free survivor. Adding abciximab to a stenting procedure decreases the incidence of myocardial infarctions at an extra cost of Euros 13,000 (95% u.l. Euros 27,000) per additional myocardial infarction-free survivor. In the subgroup of diabetics, adding abciximab improves revascularization rates as well, resulting in a cost-efficacy rate of Euros 2000 (95% u.l. Euros 25,000) per additional MACE-free survivor, with uncertainty regions indicating potential costs savings. CONCLUSION: The combination of stenting and abciximab costs about Euros 13,000 to avoid one event after PTCA. In diabetic patients the strategy may be cost-saving. PMID- 11482922 TI - Serial evaluation of perfusion defects in patients with a first acute myocardial infarction referred for primary PTCA using intravenous myocardial contrast echocardiography. AB - AIMS: To investigate whether myocardial contrast echocardiography using Sonazoid could be used for the serial evaluation of the presence and extent of myocardial perfusion defects in patients with a first acute myocardial infarction treated with primary PTCA, and specifically, (1) to evaluate safety and efficacy of myocardial contrast echocardiography to detect TIMI flow grade 0--2, (2) to evaluate the success of reperfusion and (3) to predict left ventricular recovery after 4 weeks follow-up. METHODS AND RESULTS: Fifty-nine patients underwent serial myocardial contrast echocardiography, immediately before primary PTCA (MCE1), 1 h (MCE2) and 12--24 h after PTCA (MCE3). A perfusion defect was observed in 21 of 24 patients (88%) with anterior acute myocardial infarction. All but one had TIMI flow grade 0--2 prior to PTCA. Nine of 31 patients (29%) with inferior acute myocardial infarction showed a perfusion defect and all had TIMI flow grade 0-2 prior to PTCA. Restoration of TIMI flow grade 3 was achieved in 73% of the patients by primary PTCA. A reduction in size of the initial perfusion defect of at least one segment (16 segment model) or no defect vs persistent defect in patients with anterior acute myocardial infarction was associated with improved global left ventricular function at 4 weeks; mean global wall motion score index 1.29+/-0.21 vs 1.66+/-0.31 (P=0.009). Multiple regression analysis in patients with an anterior acute myocardial infarction revealed that the extent of the perfusion defect at MCE3 was a significant (P=0.0005) independent predictor for left ventricular recovery at 4 weeks follow-up. The only other independent predictor was TIMI flow grade 3 post PTCA (P=0.007). CONCLUSION: Intravenous myocardial contrast echocardiography immediately prior to primary PTCA seems safe and is capable of detecting the presence of a perfusion defect and its subsequent dynamic changes, particularly in patients with a first anterior acute myocardial infarction. A significant reduction in size of the initial perfusion defect using serial myocardial contrast echocardiography predicts functional recovery after 4 weeks and these findings underscore the potential diagnostic value of intravenous myocardial contrast echocardiography. PMID- 11482923 TI - Peak exercise cardiac power output; a direct indicator of cardiac function strongly predictive of prognosis in chronic heart failure. AB - OBJECTIVES: This study assessed the prognostic value of peak cardiac power output, measured non-invasively during maximal cardiopulmonary exercise testing, against other exercise-derived haemodynamic variables in patients with chronic congestive heart failure. METHOD AND RESULTS: Two hundred and nineteen unselected, consecutive patients with congestive heart failure (166 men, mean (+/ SD) age of 56+/-13 years) who underwent maximal symptom limited cardiopulmonary treadmill exercise testing with non-invasive estimation of cardiac output using carbon dioxide re-breathing techniques, were followed-up for a mean period of 4.64 (4.47--4.82, 95% CI) years. Cardiac power output was calculated from the product of cardiac output and mean arterial blood pressure. All cause mortality was 12.3% (27 deaths). Peak and resting cardiac power output, peak mean arterial blood pressure, peak and resting cardiac output and peak VO(2)were all predictive of outcome on univariate analyses. Peak cardiac power output, either entered continuously or categorically with a cut-off value of 1.96 watts, was the only independent predictor of mortality (P=0.0004 for values < or >1.96 watts and P=0.001 for continuous values) using multivariate analysis. A relative risk ratio of 5.08 (1.94-13.3, 95% CI) was obtained for a cardiac power output <1.96 watts. CONCLUSION: Peak cardiac power output is an independent predictor of mortality that can be measured non-invasively using cardiopulmonary exercise testing. It can give further prognostic power to a peak VO(2)in the assessment of patients with congestive heart failure. PMID- 11482924 TI - Maintenance of sinus rhythm after electrical cardioversion of persistent atrial fibrillation; sotalol vs bisoprolol. AB - AIMS: The purpose of the study was to compare the efficacy and safety of sotalol and bisoprolol in the maintenance of sinus rhythm after electrical cardioversion of atrial fibrillation. METHODS: Patients (n=128) were randomized to sotalol (80 mg b.i.d.) or bisoprolol (5 mg x day(-1)). Patients with contraindications to beta-blockers, class III antiarrhythmic drugs or prior treatment with use of study medication for prevention of atrial fibrillation were excluded. Follow-up clinical evaluation was performed 1 day and 1 month after cardioversion and thereafter at 3-month intervals. RESULTS: There were no group differences in baseline clinical characteristics. After a follow-up of 12 months, 59% of all patients were still in sinus rhythm. The fraction remaining in sinus rhythm was calculated for the two groups by Kaplan--Meier analysis. During follow-up, 41% of patients on sotalol and 42% on bisoprolol developed atrial fibrillation (ns). In two patients (3.1%) on sotalol, life-threatening proarrhythmias (torsade de pointes tachycardias) occurred, whereas none were found in the bisoprolol group. Symptomatic bradycardias occurred in two patients on sotalol and three on bisoprolol. CONCLUSION: This study demonstrates that sotalol (160 mg x day(-1)) and bisoprolol (5 mg x day(-1)) are equally effective in maintaining sinus rhythm. Because of the side effects of sotalol, bisoprolol seems to be advantageous for maintenance of sinus rhythm after cardioversion of atrial fibrillation. PMID- 11482925 TI - Molecular analysis of connexin 40 in the familial form of atrial fibrillation. PMID- 11482926 TI - Effects of oral sotalol administration before electrical cardioversion of persistent atrial fibrillation. PMID- 11482927 TI - Over-training syndrome as a model of a coronary inflammation process? PMID- 11482929 TI - Cholesterol reduction, statins and the cytochrome P-450 system. No more recipes please. PMID- 11482932 TI - Reductions in hamster serum thyroxine levels by melatonin are not altered by changes in serum testosterone. AB - Daily melatonin injections reduce reproductive and thyroid hormones in male Syrian hamsters. The interrelationship between the decline in these hormones is not known. To explore this relationship, male Syrian hamsters were divided into four groups: castrated, implanted with testosterone (5-mm silastic implants), both treatments, or neither treatment. One-half of each group of hamsters (n = 7 or 8) were injected with melatonin (25 mg) daily at 1730 h. The other half of each group received daily vehicle injections. Ten weeks later, the hamsters were anesthetized and decapitated. Testes weights, serum testosterone, and serum thyroxine levels were measured. As expected, testes and serum testosterone levels were uniformly low in all of the melatonin-treated hamsters. All of the melatonin treated groups also had lower than normal thyroxine values irrespective of gonadal treatment. Interestingly, in the non-melatonin-treated hamsters, serum thyroxine values were decreased in the castrated group and increased in the testosterone-implanted group. These results suggest that castration can reduce serum thyroxine levels in male Syrian hamsters and that replacement of testosterone restores these levels to normal. Notably, the declines in thyroxine levels produced by daily melatonin injections were not restored by testosterone implants in castrated or intact hamsters. Therefore, melatonin-induced reductions in thyroxine are not mediated by concurrent reductions in serum testosterone levels. It appears that melatonin-induced reductions in serum thyroxine levels do not use the same mechanism as castration-induced reductions. PMID- 11482933 TI - Seasonal changes in beta-endorphin-like immunoreactivity in the olfactory system of the female catfish, Clarias batrachus (Linn). AB - In the olfactory system of the catfish Clarias batrachus, beta-endorphin-like immunoreactivity was seen in several olfactory receptor neurons (ORN) and their fiber projections extending caudally over the olfactory nerve to the olfactory bulb (OB). With beta-endorphin-like immunoreactivity as a cellular marker, the olfactory system in the female fish was investigated at different stages of its annual reproductive cycle. The reproductive cycle of the fish is divisible into four distinct phases: preparatory (February-April), prespawning (May-June), spawning (July-August), and postspawning (September-January). The gonosomatic index and the immunocytochemical profile of beta-endorphin-like immunoreactivity showed distinct changes as the fish progressed from one phase to another. In the preparatory phase, limited immunoreactivity was seen in the periphery of the bulb. However, the immunoreactivity showed a robust increase as the immunolabeled fibers extended progressively deeper into the bulb toward the mitral cell layer during the prespawning and spawning phases. Significant reduction in the immunoreactivity was noticed in the olfactory nerve layer of the fish in the postspawning phase. Several granule cells showed poor to moderate immunoreactivity during the spawning phase, although no immunoreactivity was seen in the inner cell layer during the rest of the year. The beta-endorphin-like immunoreactivity in the ORN also showed season-related changes, although these were less distinct. Whereas weak immunoreactivity confined to a few ORN was noticed in the fish collected in the preparatory phase, those in the prespawning phase showed conspicuous augmentation in immunoreactivity. During the spawning phase, the sensory layer of the olfactory epithelium showed reduced, homogenous immunoreactivity. In the postspawning phase, several ORN revealed distinct granular immunoreactivity, suggesting possibilities of de novo synthesis. These annual cyclic changes in the beta-endorphin-like immunoreactivity were consistently observed over a 30-month study period that spanned three consecutive spawning phases. The results suggest that the beta-endorphin-containing ORN, their fiber projections to the OB, and several granule cells in the inner cell layer may be involved in the processing of reproduction/reproductive behavior related signals. PMID- 11482934 TI - Differential expression of Gh1 and Gh 2 genes by competitive RT-PCR in rainbow trout pituitary. AB - The expressions of the GH1 and GH2 genes were examined by competitive RT-PCR and whole-mount in situ hybridization of pituitary of starved rainbow trout (Oncorhynchus mykiss). lambda RNA having GH primers at both 5' and 3' sites of the gene was used for the competitive RT-PCR, and thermostable reverse transcriptase produced a reasonable band when authentic RNA was examined. The amount of GH1 gene expression was significantly (P < 0.05) greater than that of GH2. Although almost the same amount of GH1 gene expression was obtained during the day, it was significantly (P < 0.05) decreased at midnight. However, there was no significant change in GH2 gene expression in the daily cycle. There were also some differences in gene expression in the pituitary: GH2 gene was more widely expressed than GH1. However, the signal intensity of the GH1 gene was greater than that of GH2. PMID- 11482935 TI - Dehydroepiandrosterone in songbird plasma: seasonal regulation and relationship to territorial aggression. AB - Many male animals are territorial in the breeding season, when plasma testosterone (T) levels are high, and nonterritorial in the nonbreeding season, when plasma T levels are basal. In contrast to this common pattern, male song sparrows (Melospiza melodia morphna) are territorial year-round, except briefly during molt. Song sparrows are highly aggressive in the nonbreeding season (autumn and winter), even though plasma T, 5 alpha-dihydrotestosterone, androstenedione (AE), and 17beta-estradiol levels are undetectable (50% product yield. The pNP half-life in ZnS nanocrystal photocatalyzed reactions was about 1.95 to 2.45 min, whereas in comparable TiO(2) reactions, the pNP half-lives were in the range of 12 to 15 min. Absorption spectra of the photocatalysis reactions suggested the decolorization of pNP without any noticeable formation of phenolic intermediates, implying a mechanism that involves a pNP ring opening via a radical mediated attack. Likewise, the degradation of AO7 was suggested to occur via an oxidative pathway involving hydroxyl radicals formed at the photocatalyst/liquid interface. Optimum conditions for AO7 degradation such as pH, photocatalyst-to-AO7 ratio, and photocatalyst surface passivation were similar to those for pNP. By demonstrating efficient mineralization of these model pollutants using mass produced ZnS nanocrystals, we hope to lay the foundations necessary for development of large-scale, field-applicable systems. Copyright 2001 Academic Press. PMID- 11482962 TI - Silylating Agents Grafted onto Silica Derived from Leached Chrysotile. AB - Silica from leached chrysotile fibers (SILO) was silanized with trialkoxyaminosilanes to yield inorganic-organic hybrids designated SILx (x=1-3). The greatest amounts of the immobilized agents were quantified as 2.14, 1.90, and 2.18 mmol g(-1) on SIL1, SIL2, and SIL3 for -(CH(2))(3)NH(2), (CH(2))(3)NH(CH(2))(2)NH(2), and -(CH(2))(3)NH(CH(2))(2)NH(CH(2))(2)NH(2) groups attached to the inorganic support. The infrared spectra for all modified silicas showed the absence of the Si-OH deformation mode, originally found at 950 cm(-1), and the appearance of asymmetric and symmetric C-H stretching bands at 2950 and 2840 cm(-1). Other important bands associated with the organic moieties were assigned to nu(as)(NH) at 3478 and nu(sym)(NH) at 3418 cm(-1). The NMR spectrum of the solid precursor material suggested two different kinds of silicon atoms: silanol and siloxane groups, between -90 and 110 ppm; however, additional species of silicon that contain the organic moieties bonded to silicon at -58 and -66 ppm appeared after chemical modification. These modified silicas showed a high adsorption capacity for cobalt and copper cations in aqueous solution, in contrast to the original SILO matrix, confirming the unequivocal anchoring of silylating agents on the silica surface. Copyright 2001 Academic Press. PMID- 11482963 TI - The Heavy-Atom Effect on the Photophysics of Aromatic Hydrocarbons in the Presence of Solid Clays. AB - The exchange of the original cation present on a Laponite clay (usually Na(+)) for heavy atoms such as Rb(+), Cs(+), and Tl(+) significantly alters the emission characteristics of some aromatic hydrocarbons (p-terphenyl, naphthalene, pyrene, and biphenyl). The increase of the atomic mass of the cation induces a decrease of the fluorescence emission simultaneous with an increase of the emission in the region of lower energies of the spectra, ascribed to the phosphorescence of those hydrocarbons. Time-resolved experiments for the pyrene-clay system showed a decrease of singlet lifetimes for the heavier atoms. Hydrocarbon aggregates were also detected from both the emission spectra and the time-resolved studies. The "excimer-like" emission showed longer lifetimes (10-25 ns) than the monomolecular hydrocarbons (1-3 ns), as already found for other similar systems. The amount of aggregates increased for the heavier cations due to the smaller surface available on the clay particles. Experiments increasing the amount of Tl(+) in samples containing a constant concentration of naphthalene allowed evaluation of the distance between the heavy atoms and the probe on the clay surface. The Perrin model treatment was used and resulted in approximately R(0)=9.2 A. Copyright 2001 Academic Press. PMID- 11482964 TI - Thermocapillary Alignment of Gas Bubbles Induced by Convective Transport. AB - The effect of a weak convective heat transfer on the thermocapillary interaction of two bubbles with an arbitrary orientation relative to an externally imposed temperature gradient is examined. Asymptotic analysis of the case of large separation distances, Z, suggests that the corrections to the bubbles' velocities are of O(Pe/Z(2)), rather than O(Pe(2)) previously found for an isolated bubble. Equal-sized bubbles are known to move with the same velocities, as if they were isolated, when heat conduction is the only transport mechanism. However, the convective transport results in a relative motion of the bubbles. The tendency of equal bubbles to line up in a plane perpendicular to the applied thermal gradient is shown analytically in the weakly nonlinear limit of small Pe numbers, and an interesting interaction behavior in the case of unequal bubbles is discussed. Copyright 2001 Academic Press. PMID- 11482965 TI - Synthesis and Surface Properties of N-Alkyl-N-methylgluconamides and N-Alkyl-N methyllactobionamides. AB - Three series of nonionic N-alkylaldonamides, N-alkyl-N-methylgluconamides (Cn MGA, Cn: n-C(10)H(21), n-C(12)H(25), n-C(14)H(29), n-C(16)H(33), and n C(18)H(37)), N-alkyl-N-methyllactobionamides (Cn-MLA, alkyl as above-mentioned), and N-oleyl-N-methylglucon/lactobionamide, were synthesized in the reaction of an appropriate N-alkyl-N-methylamine with delta-D-glucolactone and lactobionic acid, respectively. Krafft temperatures of aqueous solutions and surface properties of these surfactants at 20 degrees C, i.e., surface excess concentration, Gamma(cmc), surface area demand per molecule, A(min), efficiency in surface tension reduction, pC(20), effectiveness in surface tension reduction, Pi(cmc), critical micelle concentration, CMC, and CMC/C(20) parameter as well as standard free energies of adsorption, DeltaG degrees (ads), and of micellization, DeltaG degrees (mic), were determined. It was shown that introduction of the methyl group to the amide nitrogen increased the solubility of the surfactants, which was confirmed by their Krafft temperatures. Lactobionamides are more water soluble than gluconamides. On the other hand, the Cn-MGA surfactants are more surface active than the respective Cn-MLA ones. This observation is based on the determined adsorption and micellization parameters. The presence of one double bond in a hydrocarbon chain as in oleyl-amides increases their hydrophilic character compared with that of saturated C18 derivatives. No distinct differences were observed between the A(min) values obtained for both series studied, although they differ markedly in the size of the hydrophilic groups. Copyright 2001 Academic Press. PMID- 11482966 TI - Fluorescence Quenching of Anthracene by N,N-Diethylaniline in the Sodium Dodecyl Sulfate/Benzyl Alcohol/Water System. AB - Photoinduced electron-transfer reaction of anthracene with N,N-diethylaniline (DEA) was studied in the SDS (sodium dodecyl sulfate)/BA (benzyl alcohol)/H(2)O system. In an oil/water microemulsion, only the excited anthracene located at the interface can be quenched by DEA. In a water/oil microemulsion, this quenching reaction occurs in the BA continuous phase. Besides being the quencher of the excited anthracene, DEA could also change the system's structure. Copyright 2001 Academic Press. PMID- 11482967 TI - Formation and Characterization of Reversed Micelles Composed of Phospholipids and Fatty Acids. AB - The formation of reversed micellar systems composed of phosphatidylcholine (PC) and fatty acid was newly demonstrated by a significant increase in water content in the organic ethyl oleate phase when the micelles were prepared by the contact method. The solubilized water concentration in the reversed micellar organic phase reached 3 wt%. The new systems are expected to be used as highly biocompatible reversed micellar systems. The structure of the reversed micelles composed of PC and oleic acid was characterized by determining the water concentration and by small-angle X-ray scattering analysis. The reversed micelles composed of PC and oleic acid formed in ethyl oleate were spherical. The radius of gyration was between 30 and 50 A. The size of the reversed micelles decreased with an increase in the oleic acid concentration and was independent of the PC concentration. Experimental results indicated that the structure of the reversed micellar system was determined by the oleic acid concentration. An increase in the PC concentration caused an increase in the number of reversed micelles of the same size. These reversed micellar systems are expected to be used as solubilization media in pharmaceutical and food industries because they are not toxic. Copyright 2001 Academic Press. PMID- 11482968 TI - Substituent Effects on Binding Constants of Carotenoids to n-Heptane/AOT Reverse Micelles. AB - The absorption spectra of 6'-apo-beta-caroten-6'-ol (1), 6'-apo-beta-caroten-6' oic acid (2), and ethyl 6'-apo-beta-caroten-6'-oate (3) were analyzed in homogeneous media and in reversed micelles of AOT (sodium 1,4-bis(2-ethylhexyl) sulfosuccinate) in n-heptane. The possible solute-solvent interactions of these compounds were analyzed in pure solvents by Taft and Kamlet's solvatochromic comparison method. These carotenoids show sensitivity similar to that of medium polarity-polarizability as measured by pi*. Moreover, the absorption spectra of carotenoid 3 and to much less extent carotenoid 2 display broadening of the visible bands induced by polar solvents characteristic of carotenoids that contain a carbonyl functional group in conjugation with the carbon-carbon pi electron system. They are also sensitive to the ability of the solvent to accept protons in a hydrogen bond interaction measured by beta. This sensitivity follows the expected order: 2>1>3. In the reverse micellar system, while the spectra for 3 remain unchanged, the intensity of the absorption band characteristic of n heptane for 1 and 2 decreases as the AOT concentration increases, and a new band develops. This new band is attributed to the solute bound to the micelle interface. These changes allowed us to determine the binding constant (K(b)) between these compounds and AOT. At W(0)=[H(2)O]/[AOT]=0 the values of K(b) of 326+/-5 and 6.2+/-0.3 were found for the acid 2 and the alcohol 1, respectively. The strength of binding is interpreted considering their hydrogen-bond donor ability and the solubility in the organic pseudophase. For 1 K(b) decreases as W(0) is increased, while for 2 no variation was observed. These effects are discussed in terms of carotenoid-water competition for interfacial binding sites. Copyright 2001 Academic Press. PMID- 11482969 TI - Poly(N-vinyl-2-pyrrolidone) and 1-Octyl-2-pyrrolidinone Modified Ionic Microemulsions. AB - The influence of the nonionic polymer poly(N-vinyl-2-pyrrolidone) (PVP) in comparison to the surfactant 1-octyl-2-pyrrolidinone (OP) on the phase behavior of the system SDS/pentanol/xylene/water was studied. In both modified systems a strong increase in the water solubilization capacity was found, accompanied by a change in the spontaneous curvature toward zero. In the polymer-modified system an isotropic phase channel is formed with increasing polymer content that connects the L1 and the L2 phase. The lamellar liquid crystalline phase is destabilized in both cases. In the L1 phase the adsorption of PVP at the surface of the microemulsion droplets and the formation of a cluster-like structure is proven by several methods like (13)C NMR T(1) relaxation time measurments, zeta potential measurements, and rheology. In the L2 phase a modification of the interface of the inverse droplets is detected by a shift in the percolation boundary (conductivity) and (13)C NMR T(1) relaxation measurements. The formation of a cluster-like structure can be assumed on the basis of our rheological measurements. Copyright 2001 Academic Press. PMID- 11482970 TI - Modeling of Equilibrium Adsorption and Surface Tension of Cationic Gemini Surfactants. AB - A series of equilibrium tension models are used to evaluate the adsorption behavior of a novel class of lipoaminoacid gemini cationic surfactants, N(alpha),N(omega)-bis(long-chain N(alpha)-acylarginine)alpha,omega-dialkylamides or bis(Args). For purposes of comparison, the monomer LAM (the methyl ester of N(alpha)-lauroyl arginine) was also examined. These surfactants are of particular interest for both their low toxicity and biocompatibility. The tension models are based on the Gibbs adsorption isotherm and classified as "ionic" when the surface charge and the electric double layer are accounted for or as "pseudo-nonionic" when the surface charge is ignored. Both model predictions and fitted parameter values are evaluated with respect to physical plausibility and overall goodness of fit to the available tension and density data. In particular, the inferred values for the standard Gibbs free energy of adsorption DeltaG degrees, determined from an equilibrium constant defined on a nondimensional basis, without including artifacts due to an electrostatic contribution, are analyzed. The most reliable values of DeltaG degrees are found with the combined model to range from -110 to -120 kJ mol(-1) for the three dimers examined and -80 kJ mol( 1) for the monomer. For spacer chain lengths n=3, 6, or 9, the maximum surface area of surfactant adsorption and the maximum free energy of adsorption are observed for the surfactant with the spacer chain length of 6. Copyright 2001 Academic Press. PMID- 11482971 TI - Rheology and Permeability of Crosslinked Polyacrylamide Gel. AB - Gels produced by crosslinking polyacrylamide solutions with chromium (III) have been characterized by dynamic rheology studies. To vary the gel strength, different polymer concentrations were used, while keeping the temperature, salinity, and crosslinker concentration constant. Both the loss and storage moduli increased with the polymer concentration for this gel system. The storage modulus at the end of the gelation was used to characterize the gel strength. Steady-state water flow experiments through gel-filled capillary tubes were performed, with the aim of linking the gel strength and flow behavior. The permeability was found to be a function of the water flow rate (velocity) and polymer concentration. Two parameters were used to characterize the flow behavior, intrinsic gel permeability and elasticity index, which are each functions of the polymer concentration. However, only one parameter is needed to fully identify the flow and rheological gel properties, as the elasticity index and storage modulus are linked by a power-law relationship. The loss modulus and intrinsic permeability are correlated with the storage modulus and elasticity index, respectively. A theoretical model for this behavior linking both gel properties based on the dual domain structure was used to demonstrate that the flow and rheological behavior of the gel are indeed related and that the gel strength controls the water permeability. Implications for prediction of flow of water through gels emplaced in a porous medium are discussed. Copyright 2001 Academic Press. PMID- 11482972 TI - Investigation of Calcium Carbonate Scaling Inhibition and Scale Morphology by AFM. AB - The calcium carbonate scale inhibition by two inhibitors, polyacrylic acid (PAA) and 2-phosphonobutane-1,2,4-tricarboxylic acid (PBTCA), has been studied in two heat transfer systems: recirculating cooling water and pool boiling systems. It is found that PBTCA has a better inhibition effect than PAA under identical conditions. The inhibition effect increases with increasing fluid velocity for the cooling water system, whereas in the presence of inhibitors, the fluid velocity has less effect on the scaling behavior. When the initial surface temperature increases, the inhibition efficiency decreases. In the presence of inhibitors, the scaling behavior is insensitive to the change of surface temperature. The relationship between the inhibition effect and the fractal dimension has also been investigated. The results show that the fractal dimension is higher in the presence of inhibitors. The better the inhibition effect, the higher the fractal dimension. XRD and FTIR analyses demonstrate that for the CaCO(3) formed in the pool boiling system, the content of vaterite increases with the increase of inhibition effects. The metastable crystal forms of vaterite and aragonite are stabilized kinetically in the presence of inhibitors. The step morphology has been observed by atomic force microscopy. It is shown that the step space on the CaCO(3) surface increases in the presence of inhibitors. Moreover, with the increase in inhibition effect, both the step space and the fractal dimension increase. Step bunching is also found and discussed in this paper. Copyright 2001 Academic Press. PMID- 11482973 TI - Polydispersity and Functional Group Distribution of Dispersant Polymer: Adsorption Properties and Magnetic Paint Dispersion. AB - The quantitative analysis examining the functional group distribution of a dispersant polymer for magnetic paints is conducted by statistical estimation and adsorption experiments. The dispersant polymer contains averagely one or two functional groups on the chain, and has generally large polydispersity. By the calculation based on the random distribution of the functional group and the molecular weight, a typical design of the dispersant polymer is found to contain a significant amount of nonfunctionalized chains and highly functionalized ones. In adsorption experiments, the adsorbed amount of the polymer mass and the functional group are separately measured to determine the functional group distribution. The distribution is also evaluated by a sequential adsorption experiment, in which the chains are fractionated by the adsorption strength. Obtained experimental results agree with the calculated results. A practical method for increasing the effective chains in the paint is to make use of a preferential adsorption of the functionalized chain. Copyright 2001 Academic Press. PMID- 11482974 TI - Aggregation and Polymerization of PEG-Based Macromonomers with Methacryloyl Group as the Only Hydrophobic Segment. AB - Aggregation behavior in aqueous solution of a series of poly (ethylene glycol) (PEG)-based macromonomers with methacryloyl group as the only hydrophobic segment has been investigated using surface tension, steady-state and time-resolved fluorescence spectroscopy using pyrene as a probe, and small-angle neutron scattering techniques. The general formula of these macromonomers is CH(2)&dbond;C(CH(3))-CO-O-E(m)-CH(3), where E is the ethylene glycol unit and m=8 (ME(8)), 18 (ME(18)), 49 (ME(49)), and 120 (ME(120)). The results indicate that a macromonomer with 8 ethylene glycol units forms as an aggregate above a certain critical concentration, which can be defined as critical aggregation concentration. The observed high value of I(1)/I(3) in pyrene emission spectra at the interface of these aggregates and the inability to scatter a neutron beam by these aggregates indicate that the hydrophobic cluster formed by this macromonomer is remarkably solvated. ME(18) has a tendency to aggregate but others do not form any hydrophobic cluster. The homopolymerization behaviors of these macromonomers in an aqueous medium at 70 degrees C are consistent with these possibi- lities. Copyright 2001 Academic Press. PMID- 11482975 TI - Effects of Nonionic and Mixed Cationic-Nonionic Micelles on the Rate of Alkaline Hydrolysis of 4-Nitrophthalimide. AB - Pseudo-first-order rate constants (k(obs)) for alkaline hydrolysis of 4 nitrophthalimide show a monotonic decrease with increase in [C(12)E(23)](T) (total concentration of Brij 35) at constant [CH(3)CN] and [NaOH]. This micellar effect is explained in terms of a pseudophase micelle model. The rate of hydrolysis becomes too slow to monitor at [C(12)E(23)](T)>/=0.03 M in the absence of cetyltrimethylammonium bromide (CTABr) and at [C(12)E(23)](T)>/=0.04 M in the presence of 0.006-0.02 M CTABr at 0.01 M NaOH. The plots of k(obs) versus [C(12)E(23)](T) show minima at 0.006 and 0.01 M CTABr, while such a minimum is not visible at 0.02 M CTABr. Copyright 2001 Academic Press. PMID- 11482976 TI - A Model for Hydration Interactions between Apoferritin Molecules in Solution. AB - We have studied how non-DLVO forces between molecules of the globular protein apoferritin in solution affect its osmotic second virial coefficient. A model explaining the effects of the solution ionic strength and pH on the interprotein interaction is developed, to give a physical interpretation of recently published experimental findings showing that the second virial coefficient of the protein apoferritin, supported by acetate buffer, goes through a minimum as a function of ionic strength. At low ionic strengths, the apoferritin second virial coefficient initially decreases with increasing sodium ion concentration, as DLVO theory predicts. However, non-DLVO hydration forces due to overlapping of the Stern layers of the protein molecules increase the second virial coefficient with further increase of sodium ion concentration, again as found experimentally at higher ionic strengths. The non-DLVO effect arises from ionic exchange between hydrogen and sodium ions at the protein surface. An adsorption shell of hydrated sodium ions forms around the protein molecules with increasing buffer concentration. Copyright 2001 Academic Press. PMID- 11482977 TI - Adverse event reporting: essential for science and public trust. PMID- 11482978 TI - Challenges and strategies for cystic fibrosis lung gene therapy. AB - Gene replacement therapy represents an interesting new approach for the treatment of cystic fibrosis (CF) lung disease. Basic research suggests that CF gene therapy is feasible, but major technological challenges must be addressed before clinical applications are likely to succeed. Therapeutic genes can be delivered to and expressed in human airways, but the number of cells expressing the transgene is relatively low. The inefficiency of gene delivery is largely attributable to the remarkable defenses of human airways. Maintaining long-term transgene expression in airway cells is also a significant obstacle. Recent advances have been made in the development of vectors, expression cassettes, and delivery techniques for enhancing airway gene transfer and expression. These advances have the potential to improve the efficiency of lung gene therapy and to achieve clinical benefits for CF patients in the future. PMID- 11482979 TI - Clinical gene transfer: education is the key. PMID- 11482980 TI - Development of an MFG-based retroviral vector system for secretion of high levels of functionally active human BMP4. AB - We sought to develop a retroviral vector system that would produce secretion of high levels of bone morphogenetic protein (BMP)-4 by optimizing the expression construct and developing an improved retroviral vector. Replacement of the propeptide domain of BMP4 with that of BMP2 increased the secretion level of mature BMP4 protein in transduced cells. The intact BMP2 pro-peptide sequence was essential, as deletion of a small part of the propeptide sequence of BMP2 from the BMP2/4 hybrid construct diminished BMP4 expression and secretion. Addition of a hemaglutinin tag to the carboxy terminus of BMP4 abolished the bioactivity of secreted BMP4. Transduction of rat marrow stromal cells (and fibroblasts) with an MFG-based retroviral vector pseudotyped with VSV-G envelope containing this BMP2/4 hybrid expression construct led to secretion of very high levels of mature BMP4 in conditioned medium (up to 1 microg/10(6) cells/24 hours). The secreted BMP4 was biologically active, as it induced alkaline phosphatase expression in C2C12 cells. The transduced rat marrow stromal cells expressing mature BMP4 induced de novo ectopic bone formation in syngenic immune-competent rats. We have developed an MFG-based retroviral vector system that causes secretion of high levels of functionally active human BMP4 protein. PMID- 11482981 TI - Messenger RNA repair and restoration of protein function by spliceosome-mediated RNA trans-splicing. AB - The functional repertoire of the human genome is amplified by the differential assortment of exons. Spliceosome-mediated RNA trans-splicing can mobilize these packets of genetic information to reprogram mRNAs. In principle, this process could repair defective transcripts in loss-of-function genetic disorders in humans. We developed a tractable lacZ repair system to serve as a model for these genetic disorders. Targeted pre-trans-splicing RNA molecules efficiently and specifically repaired mutated lacZ transcripts and restored enzymatic activity in human cells. The development of this model confirms the potential for spliceosome mediated RNA trans-splicing in genetic repairs and provides a powerful tool for rational design and in vitro evolution of pre-trans-splicing molecules. PMID- 11482982 TI - In utero AAV-mediated gene transfer to rabbit pulmonary epithelium. AB - In utero intra-amniotic administration of adeno-associated virus (AAV) for treatment of cystic fibrosis (CF) has the potential to be an efficient way to target the rapidly dividing undifferentiated cells of the fetal pulmonary epithelium, while simultaneously treating other tissues involved in CF (such as the intestines), but has never before been studied. Intra-amniotic administration of 1x10(12) particles of AAV-luciferase vector to 110 fetal rabbits at 24-25 days gestation resulted in transgene expression in amniotic membranes, trachea, and pulmonary epithelium. The highest level of transgene expression was found in amniotic membranes. Transgene expression peaked in the lungs 10 days after vector delivery, decreased at day 17, and was no longer detectable after 24 days. The number of pulmonary cells transduced was approximately 1 in 500 and immunohistochemical analysis showed expression in varying cell types, including alveolar cells. Transgene expression was not detected in fetal rabbit intestines, skin or liver, nor in maternal ovaries or liver. Intra-amniotic administration of AAV does not result in the tissue inflammation and fetal loss previously documented with in utero adenoviral administration, and results in high levels of transgene expression in amniotic membranes with lower levels in fetal pulmonary epithelium. PMID- 11482983 TI - Epstein-Barr virus/human vector provides high-level, long-term expression of alpha1-antitrypsin in mice. AB - We have constructed plasmid DNA vectors that contain Epstein-Barr virus (EBV) sequences and the human gene (SERPINA1) encoding alpha1-Antitrypsin (AAT). We demonstrate that a plasmid carrying the full SERPINA1 on a 19-kb genomic fragment and the EBV gene EBNA1 and its family of repeats binding sites undergoes efficient extrachromosomal replication in dividing mammalian tissue culture cells. Therefore, use of a whole genomic therapeutic gene to provide both replication and gene expression may be an effective gene therapy vector design, if the target cells are dividing. The efficacy of this same vector for expression of AAT in vivo in the nondividing cells of mouse liver was determined by hydrodynamic injection of naked plasmid DNA by means of the tail vein. A single injection of an EBV/genomic SERPINA1 vector provided >300 microg/ml of AAT, which approached normal plasma levels and persisted for the >9-month duration of the experiment. These data exceed most previously reported values, probably due to sequences in the genomic DNA that resist silencing of gene expression, possibly in combination with favorable effects on expression provided by the EBV sequences. These results demonstrate that plasmid DNA with the correct cis-acting sequences can provide in vivo long-term expression of protein at high levels that are therapeutically relevant for gene therapy. PMID- 11482984 TI - Intratumoral delivery of p2CMVmIL-12 using water-soluble lipopolymers. AB - Our objective was to design a water-soluble lipopolymer (WSLP) and an interleukin 12 (IL-12) expression plasmid for enhanced delivery of the IL-12 gene. We synthesized WSLP using branched polyethylenimine (PEI) of 1800 Da and cholesteryl chloroformate, and constructed p2CMVmIL-12, encoding the IL-12 subunits p35 and p40, each under the transcriptional control of a separate cytomegalovirus (CMV) promoter. The percentage of cholesterol conjugated to PEI was about 47% and the average molecular weight of WSLP was approximately 2000 Da. The mean particle size of WSLP/p2CMVmIL-12 complexes formulated in 5% glucose was 26 to 62 nm and xi potential was 8 to 60 mV. The WSLP/p2CMVmIL-12 complexes were nontoxic to CT 26 colon carcinoma cells at the N/P ratio (nitrogen atoms of WSLP/phosphate of plasmid DNA) of 20 and below; PEI25000/pDNA complexes were highly toxic. WSLP/p2CMVmIL-12 complexes demonstrated higher transfection in CT-26 cells compared with the DNA formulations prepared using PEI of molecular weights 1800, 10,000 and 25,000 Da. Transfection efficiency increased with an increase in N/P ratios from 5 to 15, then there was no significant increase in transfection up to the N/P ratio of 30/1. There was an increase in the level of IL-12 when free or complexed p2CMVmIL-12 was compared with free or complexed pIRESmIL-12 in which the p35 and p40 subunits were linked to the internal ribosome entry sites (IRES). At 48 hours post-injection of WSLP/p2CMVmIL-12 complexes into BALB/c mice bearing CT-26 subcutaneous tumors, the levels of IL-12, IFN-gamma, and nitric oxide (NO) in the supernatant of the cultured tumors were higher for the WSLP/p2CMVmIL-12 complexes than for the naked p2CMVmIL-12, WSLP, and 5% glucose injected groups. There was a significant improvement in the survival rate and the inhibition of tumor growth after a single injection of WSLP/p2CMVmIL-12 complexes. We have designed an effective, nontoxic WSLP and an IL-12 expression plasmid with two CMV promoters. PMID- 11482985 TI - Dose-dependent effects of central leptin gene therapy on genes that regulate body weight and appetite in the hypothalamus. AB - We have examined the dose-dependent effects and central action of intraventricular administration of a recombinant adeno-associated virus encoding rat leptin (rAAV-leptin) in suppressing body weight (BW) gain in adult female rats. A low dose of rAAV-leptin (5x10(10) particles) suppressed weight gain (15%) without changing daily food intake (FI), but a twofold higher dose decreased BW by 30% along with a reduction in daily FI. Reduced BW was due to a loss in body adiposity because serum leptin was reduced. Serum insulin levels were decreased (96%) by only the high dose along with a slight reduction in glucose. Uncoupling protein-1 (UCP-1) mRNA expression in brown adipose tissue (BAT), reflecting energy expenditure through thermogenesis, was upregulated to the same magnitude by the two rAAV-leptin doses. We analyzed by in situ hybridization the expression in the hypothalamus of genes encoding the appetite-regulating neuropeptides. Only the high dose decreased expression of neuropeptide Y (NPY), the orexigenic peptide, and increased proopiomelanocortin (POMC), precursor of the an orexigenic peptide, alpha-MSH. Our studies show for the first time that increased availability of leptin within the hypothalamus through central leptin gene therapy dose-dependently decreases weight gain, adiposity, and serum insulin by increasing energy expenditure and decreasing FI. The decrease in FI occurs only when NPY is reduced and alpha-MSH is increased in the hypothalamus by the high dose of rAAV-leptin. Delivery of the leptin gene centrally through rAAV vectors is a viable therapeutic modality for long-term control of weight and metabolic hormones. PMID- 11482986 TI - Elimination of the truncated message from the herpes simplex virus thymidine kinase suicide gene. AB - Introduction of the Herpes simplex virus thymidine kinase (HSV-tk) gene into target cells renders them susceptible to killing by ganciclovir (GCV). We are studying the use of HSV-tk-transduced T lymphocytes in the context of hematopoietic stem cell transplantation. We have previously shown, in vitro and in vivo, the occurrence of transduced cells resistant to GCV due to a deletion within HSV-tk. This deletion, a consequence of the presence of cryptic splice donor and acceptor sites, originates in the retroviral producer cell. Here we adopt two different methods that introduce third-base degenerate changes at the cryptic splice sites and so prevent splicing. Consequently, the HSV-tk protein is unaltered and the sensitivity of the target cells to GCV is preserved. The use of this mutated HSV-tk should reduce the likelihood of the development of resistant genetically modified cells during clinical trials. PMID- 11482987 TI - Production and neurotropism of lentivirus vectors pseudotyped with lyssavirus envelope glycoproteins. AB - We investigated the production efficiency and the gene transfer capacity in the central nervous system of HIV-1-based vectors pseudotyped with either the G protein of the Mokola lyssaviruses (MK-G), a neurotropic virus causing rabies disease, or the vesiculo-stomatitis G protein (VSV-G). Both envelopes induced syncitia in cell cultures. They were incorporated into vector particles and mature virions were observed by electron microscopy. Vector production was two- to sixfold more efficient with VSV-G than with MK-G. For equivalent amounts of physical particles, vector titration was 5- to 25-fold higher with VSV-G than with MK-G pseudotypes on cultured cells, and in vivo gene expression in mouse brain was more intense. Thus, VSV-G pseudotypes were produced more efficiently and were more infectious than MK-G pseudotypes. Tropism for brain cells was analyzed by intrastriatal injections in rats. Both pseudotypes preferentially transduced neurons (70-90% of transduced cells). Retrograde axonal transport was investigated by instilling vector suspensions in the rat nasal cavity. Both pseudotypes were efficiently transported to olfactive neuron bodies. Thus, although coating HIV-1 particles with rabdhovirus envelope glycoproteins enables them to enter neuronal cells efficiently, pseudotyping is not sufficient to confer the powerful neurotropism of lyssaviruses to lentivirus vectors. PMID- 11482988 TI - Toxicity assessment of intratumoral injection of the herpes simplex type I thymidine kinase gene delivered by retrovirus in patients with refractory cancer. AB - Introduction of the herpes simplex type I thymidine kinase (HSV-TK) gene into tumor tissue, followed by ganciclovir, initiates a phosphorylation cascade that induces formation of a toxic ganciclovir triphosphate. Animal trials suggest that this ganciclovir triphosphate has antitumor activity. Here we report application of the HSV-TK transfection approach using a retroviral construct. Sixteen patients (median age 61.5 years) with refractory carcinoma (13 melanoma, 1 breast cancer, 1 nonsmall-cell lung cancer, and 1 osteogenic sarcoma) received intratumoral injection of HSV-TK retroviral vector at escalating doses (0.2x10(7) cfu per injection x 5 daily doses) and we evaluated them for toxicity and activity. We observed grade III pain associated with cellulitis in one patient following injection. Analysis of blood samples drawn between 3 and 28 weeks from 14 patients for replication-competent retrovirus by PCR analysis of the amphotrophic envelope revealed no replication-competent retrovirus. We injected 21 lesions. We identified no tumor responses of the injected lesions. Of 13 patients with advanced melanoma, 6 survived over one year. Thus, injection of retroviral delivered HSV-TK in patients with refractory cancer was well tolerated. PMID- 11482989 TI - Cholesterol screening among children and their parents. AB - BACKGROUND: The Coronary Artery Risk Detection in Appalachian Communities (CARDIAC) project is designed to test the hypothesis that universal cholesterol screening of prepubertal schoolchildren is effective in identifying children and their parents at risk of developing premature coronary heart disease (CHD) in a high-risk rural population. METHODS: Seven hundred nine fifth-grade schoolchildren from seven rural Appalachian counties participated in a school based cholesterol screening program. Family history of premature CHD, anthropometric and blood pressure measurement, tobacco smoke exposure, dietary history, and physical activity levels were collected. RESULTS: One-fourth (174) of the children were "presumptively" dyslipidemic upon measurement of nonfasting finger-stick blood cholesterol (FSC). Subsequent fasting lipid profile obtained for 63 of these children and 79 of their parents confirmed the presence of dyslipidemia in 37 children (59%) and 52 parents (66%). Among confirmed dyslipidemic children, family history was not a good predictor of dyslipidemia (sensitivity 21.6%). FSC levels were significantly correlated with fasting total cholesterol of children and their parents. CONCLUSIONS: Universal nonfasting FSC screening of prepubertal schoolchildren is effective in identifying dyslipidemic children and their parents, whereas family history has low sensitivity in predicting children with elevated blood cholesterol concentrations. PMID- 11482990 TI - Premenstrual syndrome. a message from preventive medicine and your physician. PMID- 11482991 TI - Breast cancer screening among American Samoan women. AB - BACKGROUND: Little is known about breast cancer screening practices or predictors of age-specific screening for Samoan women. METHODS: Through systematic, random sampling procedures, we identified and interviewed 720 adult (> or =30 years) Samoan women residing in American Samoa, Hawaii, and Los Angeles. Multivariate logistic regressions were performed to determine independent predictors for recent age-specific screening. RESULTS: Only 55.6% of women (> or =30 years) had ever had a CBE and 32.9% of women (> or =40 years) had ever had a mammogram. Furthermore, only 24.4 and 22.4% of Samoan women (> or =40 years) residing in Hawaii and Los Angeles, respectively, had an age-specific mammogram within the prior year. Independent predictors of age-specific CBE screening included age, education, health insurance, ambulatory visit, and being a resident of Hawaii or Los Angeles; those for mammography included ambulatory visit and awareness of screening guidelines. CONCLUSION: Population-based estimates of age-specific breast cancer screening among Samoan women are lower than the national objectives and those reported for other minorities. Targeted efforts that address doctor patient communication on preventive behavior, improved access to health care services (especially in American Samoa), and focused educational awareness programs are needed to improve the dismal screening rates observed in this indigenous population. PMID- 11482992 TI - Can sedentary adults accurately recall the intensity of their physical activity? AB - BACKGROUND: Physical activity, in particular vigorous activity (i.e., > or =6 METs), lowers mortality from chronic diseases such as cardiovascular disease (CVD). The 7-Day Physical Activity Recall (PAR), a self-administered activity log (LOG), and heart rate monitoring (HR) were used to quantify activity patterns among sedentary adults. We hypothesized that individuals in this population could accurately estimate the duration, but not the intensity, of their activity. METHODS: Sedentary adults (n = 94, 47.8 +/- 7.1 years) completed two PARs 1 week apart and underwent HR monitoring while completing a LOG for 1 day during the PAR assessment interval. RESULTS: The relationship between PARs (kcal. kg(-1). day( 1) ) was significant (r = 0.80, 95% CI 0.68-0.87) among individuals (n = 63) reporting "typical" activity patterns and among all individuals (n = 94) reporting "typical" and "not typical" activity patterns combined (r = 0.44, 95% CI 0.26-0.59). Quantity of moderate activity was greater (P = 0.0001) on PAR and LOG compared to that measured by HR. Quantity of hard (vigorous) activity was also greater (P = 0.019) on LOG compared to that measured by HR. CONCLUSIONS: Sedentary adults tend to overestimate the intensity of their activity, specifically for moderate activity. Furthermore, the aerobic capacity of our sedentary adult sample (about 7.3 METs) suggests that the definition of a threshold intensity level of activity necessary to reduce mortality from CVD should be reexamined, because a value of > or =6 METs appears to be too high in this population. PMID- 11482993 TI - Cross-sectional measurement of nutrient intake among adolescents in 1996. AB - BACKGROUND: Many components are involved in an assessment of diet and health among youth. To address these issues and document the major contributions of fat, fiber, vitamins, and minerals to the diet, we analyzed baseline cross-sectional data from a cohort of 16,882 youth. METHODS: Cross-sectional analyses were conducted on the Growing Up Today Study dietary data reported by 16,882 9- to 14 year-olds in 1996. The adolescent food frequency questionnaire was used to assess this age group's eating habits. Arithmetic means and standard deviations were calculated on energy-adjusted and unadjusted data. RESULTS: Mean intake (including vitamin/mineral supplementation) for all nutrients met 100% of the RDAs, except calcium for girls. Comparing the servings of foods with the USDA Food Pyramid, neither boys nor girls met recommended number of servings except for dairy. Overweight participants consumed fewer kilocalories and lower levels of nutrients than their nonoverweight peers. CONCLUSION: These cross-sectional data from 1996 indicate that this cohort is consuming foods and nutrients comparable with national data of less fat and more carbohydrates in their diet. Overweight participants have similar dietary patterns except for total energy. The cohort's diet (with vitamin/mineral supplementation) is meeting the RDAs, but actual foods consumed suggest a lack of balance in the diet. PMID- 11482994 TI - Religious preference, church activity, and physical exercise. AB - BACKGROUND: Utah has the highest percentage of physically active adults in the United States. It also has a high percentage of religiously active people. About 90% of the adult population has a religious preference and 62% of those individuals attend church weekly. This study evaluates the relationship between religious preference, church attendance, and physical activity. METHODS: Analysis is based on 6,188 adult respondents ages 18 years and older to a cross-sectional random telephone survey involving 12 health districts in the state of Utah from June 1 to August 31, 1996. RESULTS: Within religious groups, those attending church weekly were more likely to exercise than individuals attending church less than weekly. About 70% of Utah consists of members of the Church of Jesus Christ of Latter-day Saints (LDS). LDS attending church less than weekly were least physically active in Utah. The percentages of those exercising vigorously at least 20 min three times per week were 52.3% (LDS who attend church weekly), 44.5% (LDS who attend church less than weekly), 57.6% (non-LDS who attend church weekly), 54.3% (non-LDS who attend church less than weekly), and 53.1% (no religious preference) (P = 0.0070). When adjustment was made for age, smoking status, education, marital status, and general health, differences in exercise between church activity levels within religious groups became insignificant. Smoking and general health have the greatest influences on the relationship between religious preference, church attendance, and exercise. LDS were significantly less physically active than non-LDS in the adjusted model. CONCLUSIONS: Although differential smoking levels and general health status explained differences between church activity levels within religious groups, lower levels of exercise among LDS versus non-LDS could not be explained and require further study. PMID- 11482995 TI - Predictors of smoking reduction and cessation in a cohort of danish moderate and heavy smokers. AB - BACKGROUND: The aim of this study was to examine the extent and gender distribution of unassisted tobacco reduction and cessation in a cohort of moderate and heavy smokers and to identify possible predictor variables associated with these changes in smoking behavior. METHODS: This was a prospective population study of 3,791 moderate and heavy smokers, 15 g tobacco/day or more, who were enrolled in the Copenhagen City Heart Study in 1976 1978 and attended a reexamination 5 years later. Data on smoking behavior were collected at baseline and follow-up. Smoking reduction was defined as a decrease in mean daily tobacco consumption of 10 g or more. Using multivariate logistic regression, subjects who reported reduced smoking or who reported smoking cessation were compared with subjects who continued the habit unchanged. RESULTS: After 5 years 13% of the men and 9% of the women had reduced their tobacco consumption, and 9 and 7%, respectively, had quit altogether. Smoking reduction was strongly associated with high tobacco consumption (25+ g/day) at baseline and also with severely impaired lung function (FEV(1) <50% predicted) and overweight (BMI >25). Predictors of smoking cessation included impaired lung function and a tobacco consumption of 15-24 g/day. Additional determinants of smoking reduction and cessation such as inhalation habits and sociodemographic variables differed by gender. CONCLUSIONS: Several predictors of smoking reduction and cessation were identified, indicating that these subgroups of smokers differ substantially from continuing smokers. This should be taken into account when assessing potential health benefits from these changes in smoking behavior. PMID- 11482996 TI - Aids to quitting tobacco use: how important are they outside controlled trials? AB - BACKGROUND: Although there is good evidence that several pharmacotherapies and counseling can effectively facilitate smoking cessation, there is little information about the use or effectiveness of these or any other quit aids outside of controlled trials. METHODS: A mailed survey with phone follow-up documented the use of various quit aids among 3,122 health plan members who smoke. A multilevel statistical modeling technique controlled for potentially confounding variables. RESULTS: Nearly half (1,513) of these smokers reported a quit attempt during the preceding 6 months. Although 1,036 (33.2%) reported using some type of aid to quitting, primarily nicotine products or bupropion, 10-26% of these "users" did not report an actual quit attempt. Ninety percent of the medication users had a personal cost, averaging $53-$87. Fully 26.9% of those reporting a quit without any type of aid quit for at least 7 days. This rate equals that of users of all types of aids except for nicotine patches and bupropion, both of which had associated 7 or more day quit rates of about 46% (95% CI 39.3-52.2). CONCLUSIONS: Pharmacotherapeutic quit aids are being widely used, even in the absence of significant insurance coverage. PMID- 11482997 TI - Laws restricting self-service tobacco displays: will they help? PMID- 11482998 TI - Glycosylated and phosphorylated proteins--expression in yeast and oocytes of Xenopus: prospects and challenges--relevance to expression of thermostable proteins. AB - Phosphorylation and glycosylation are important posttranslational events in the biosynthesis of proteins. The different degrees of phosphorylation and glycosylation of proteins have been an intriguing phenomenon. Advances in genetic engineering have made it possible to control the degree of glycosylation and phosphorylation of proteins. Structural biology of phosphorylated and glycosylated proteins has been advancing at a much slower pace due to difficulties in using high-resolution NMR studies in solution phase. Major difficulties have arisen from the inherent mobilities of phosphorylated and glycosylated side chains. This paper reviews molecular and structural biology of phosphorylated and glycosylated proteins expressed in eukaryotic expression systems which are especially suited for large-scale production of these proteins. In our laboratory, we have observed that eukaryotic expression systems are particularly suited for the expression of thermostable light-activated proteins, e.g., bacteriorhodopsins and plastocyanins. PMID- 11482999 TI - High-yield expression and purification of human interferon alpha-1 in Pichia pastoris. AB - For several years, interferon alpha-1, also known as interferon alpha-D, has been studied for treatment of various viral diseases, such as hepatic fibrosis caused by hepatitis B, herpes simplex virus keratitis, and bovine respiratory diseases in calves. Currently, recombinant human interferon alpha-D (rHuIFNalphaD) is expressed intracellularly in Escherichia coli or secreted by Bacillus subtilis and Saccharomyces cerevisiae. In this report, we describe the process of obtaining a relatively high-yield secretion of biologically active recombinant rHuIFNalphaD using the Pichia pastoris system. The process produced as high as 0.7 mg of purified protein per 20 ml of shake culture of rHuIFNalphaD with better bioactivity than the commercially available rHuIFNalphaD molecule produced in E. coli. PMID- 11483000 TI - Optimization of a thermostable lipase from Bacillus stearothermophilus P1: overexpression, purification, and characterization. AB - An expression library was generated from a partial NcoI and HindIII digest of genomic DNA from the thermophilic bacterium, Bacillus stearothermophilus P1. The DNA fragments were cloned into the expression vector pQE-60 and transformed into Escherichia coli M15[EP4]. Sequence analysis of a lipase gene showed an open reading frame of 1254 nucleotides coding a 29-amino-acid signal sequence and a mature sequence of 388 amino acids. The expressed lipase was isolated and purified to homogeneity in a single chromatographic step. The molecular mass of the lipase was determined to be approximately 43 kDa by SDS-PAGE and mass spectrometry. The purified lipase had an optimum pH of 8.5 and showed maximal activity at 55 degrees C. It was highly stable in the temperature range of 30-65 degrees C. The highest activity was found with p-nitrophenyl ester-caprate as the synthetic substrate and tricaprylin as the triacylglycerol. Its activity was strongly inhibited by 10 mM phenylmethanesulfonyl fluoride and 1 hexadecanesulfonyl chloride, indicating that it contains a serine residue which plays a key role in the catalytic mechanism. In addition, it was stable for 1 h at 37 degrees C in 0.1% Chaps and Triton X-100. PMID- 11483001 TI - Overexpression and purification of Helicobacter pylori flavodoxin and induction of a specific antiserum in rabbits. AB - Flavodoxin from the gastric pathogen Helicobacter pylori has been shown to be the electron acceptor of the essential pyruvate-oxidoreductase enzyme complex and proposed to be involved in the pathogenesis of gastric MALToma. In order to obtain a sufficient amount for biochemical and structural studies, we overexpressed the protein either with a C-terminal His(6) -tag or as a fusion protein upstream of intein- and chitin-binding domains. With both expression systems we succeeded at purifying soluble and functional flavodoxin containing the cofactor FMN. When expressing with a His(6) -tag, we purified approximately 20 mg flavodoxin per liter of bacterial culture, while expression as an intein CBD fusion protein with autocatalytic removal of the intein-CBD part rendered only approximately 1 mg of purified flavodoxin per liter of bacterial culture. Expressed as an intein-CBD fusion protein, flavodoxin copurified with a C terminal degradation product, which was not observed for expression with a His(6) -tag. However, we were able to obtain protein crystals suited for X-ray structure determination from flavodoxin expressed as an intein-CBD fusion protein, but not from flavodoxin expressed with a C-terminal His(6) -tag. We further report the induction of a rabbit antiserum specific for H. pylori flavodoxin. PMID- 11483002 TI - Expression, purification, and functional characterization of the serine protease inhibitor neuroserpin expressed in Drosophila S2 cells. AB - Neuroserpin (NS) is a serine protease inhibitor (or serpin) that is widely expressed in the developing and adult nervous systems. It has been implicated in the regulation of proteases involved in processes such as synaptic plasticity, neuronal migration, and axogenesis. To aid in the characterization of this new serpin we have established a high-level expression system in Drosophila S2 cells and developed a purification strategy to obtain neuroserpin for functional studies. Suspension cultures of S2-NS cells secreted recombinant neuroserpin into the medium. High-level expression was maintained when the cells were switched to a nonselection serum-free medium for 3-4 days to facilitate protein purification. Recombinant neuroserpin was purified by sequential chromatography on Macroprep ceramic hydroxyapatite, Type I, POROS HQ20, Resource Q, and Superdex 75 HR 10/30 media. Two secreted forms of neuroserpin were observed with molecular weights of approximately 49 and approximately 50 kDa which may represent alternative glycosylation at three putative N-linked glycosylation sites. Amino acid sequence analysis indicated three NH(2)-terminal sequences. The major sequence was generated by cleavage at the Gly(18)-Ala(19) bond consistent with removal of an 18-amino-acid signal peptide. Two further sequences were identified each with one fewer amino acids at the NH(2)-terminus. All three NH(2)-terminal sequences were also identified by mass spectrometric analysis of neuroserpin following trypsin digestion. Mass spectrometry also confirmed the protein had an intact carboxyl terminus while complex formation assays indicated the inhibitor was functionally active. In summary, Drosophila S2 cells offered a nonlytic stable expression system for the continual production of neuroserpin in high-density suspension cultures. PMID- 11483003 TI - Expression of C1 esterase inhibitor by the baculovirus expression vector system: preparation, purification, and characterization. AB - C1 esterase inhibitor (C1INH) is an important regulator of the classical complement pathway. Hereditary deficiency of C1INH causes angioedema of the skin, gut, and respiratory tissues that may be fatal. C1INH replacement therapy may be lifesaving for patients with this disorder. The objective of this study was to evaluate the use of the baculovirus expression vector system for mass producing biologically active human recombinant (rC1INH). A recombinant baculovirus was constructed coding the human native (nC1INH) sequence under control of the polyhedrin promoter. Spodoptera frugiperda Sf-9 insect cells were infected with this recombinant baculovirus in a medium-scale (10-L) bioreactor to produce rC1INH with a specific activity of 45 U/mg. Purification of rC1INH from the culture harvested at 60 h postinfection yielded 5.9 microg rC1INH/mL supernatant of a 75-kDa product with a specific activity of 31,000 U/mg purified rC1INH compared to 71,000 U/mg purified nC1INH from human serum using the same procedure. This rC1INH was about 25 kDa smaller than nC1INH, suggesting that Sf-9 cells express underglycosylated rC1INH. Glycan analysis showed that both N-glycan and O-glycan chains were present in rC1INH. The N-glycan chains, released using PNGaseF and fluorescently labeled, were analyzed using exoglycosidase treatment and capillary electrophoresis. Their high-mannose structure was consistent with the known failure of the insect cell glycosylation pathway to afford the fully elaborated biantennary structures found on human native nC1INH. PMID- 11483004 TI - Expression of a highly toxic protein, Bax, in Escherichia coli by attachment of a leader peptide derived from the GroES cochaperone. AB - Expression of the human apoptosis modulator protein Bax in Escherichia coli is highly toxic, resulting in cell lysis at very low concentrations (Asoh, S., et al., J. Biol. Chem. 273, 11384-11391, 1998). Attempts to express a truncated form of murine Bax in the periplasm by using an expression vector that attached the OmpA signal sequence to the protein failed to alleviate this toxicity. In contrast, attachment of a peptide based on a portion of the E. coli cochaperone GroES reduced Bax's toxicity significantly and allowed good expression. The peptide, which was attached to the N-terminus, included the amino acid sequence of the mobile loop of GroES that has been demonstrated to interact with the chaperonin, GroEL. Under normal growth conditions, expression of this construct was still toxic, but generated a small amount of detectable recombinant Bax. However, when cells were grown in the presence of 2% ethanol, which stimulated overproduction of the molecular chaperones GroEL and DnaK, toxicity was reduced and good overexpression occurred. Two-dimensional gel electrophoresis analysis showed that approximately 15-fold more GroES-loop-Bax was produced under these conditions than under standard conditions and that GroEL and DnaK were elevated approximately 3-fold. PMID- 11483005 TI - Cloning and overexpression in soluble form of functional shikimate kinase and 5 enolpyruvylshikimate 3-phosphate synthase enzymes from Mycobacterium tuberculosis. AB - Tuberculosis (TB) resurged in the late 1980s and an estimated 1.87 million people died of TB in 1997. The reemergence of tuberculosis as a public health threat, the high susceptibility of HIV-infected persons, and the proliferation of multidrug-resistant strains have created a need to develop new antimycobacterial agents. The existence of a shikimate pathway has been predicted by the determination of the genome sequence of Mycobacterium tuberculosis. The M. tuberculosis aroK-encoded shikimate kinase and aroA-encoded 5 enolpyruvylshikimate 3-phosphate (EPSP) synthase were cloned and the enzymes overexpressed in soluble form. Overexpression was achieved without isopropyl beta d-thiogalactoside induction, and cells grown to stationary phase yielded approximately 30% of target proteins to total soluble cell proteins. Enzyme activity measurements using coupled assays demonstrated that there was a 328-fold increase in specific activity for shikimate kinase and 101-fold increase for EPSP synthase. PMID- 11483006 TI - Expression of the myc/His-tagged human peptide transporter hPEPT1 in yeast for protein purification and functional analysis. AB - The human intestinal peptide transporter hPEPT1 has been expressed in the yeast Pichia pastoris using the promoter of the glyceraldehyde-3-phosphate dehydrogenase gene. A myc-epitope fused to a polyhistidine-tag was introduced at the C-terminus of hPEPT1 for ease of detection and purification. Yeast cells transformed with tagged hPEPT1 exhibited 30-fold increased dipeptide uptake compared to control cells with a substrate specificity and pH dependence similar to the native transporter. The tagged hPEPT1 protein was detected in crude membrane fractions of Pichia cells with an apparent molecular mass of 66 kDa and an expression level of approximately 64 pmol/mg membrane protein. These studies demonstrate that tagged hPEPT1 can be expressed functionally in P. pastoris with unaltered phenotypical characteristics allowing the yeast cells to be used for functional analysis such as screening for compounds utilizing the peptide transporter for absorption in the human intestine. Moreover, recombinant hPEPT1 can now easily be detected for further purification purposes using immobilized metal-affinity chromatography. PMID- 11483007 TI - Human aromatase in high yield and purity by perfusion chromatography and its characterization by difference spectroscopy and mass spectrometry. AB - Expression of human cytochrome P450 aromatase (CYP19A1, aromatase) was accomplished at a high level using a baculovirus expression system in an insect cell suspension culture. Using the relatively new chromatographic technique of perfusion chromatography, a very rapid procedure for purification of the protein from solubilized cells was developed. At extraordinary flow rates of between 3 and 9 column volumes per minute, all chromatographic procedures could be performed, including setup, equilibration, and column regeneration steps, in less than 2 h, not including brief dialysis periods. Total yields were 40-52% and resulted in preparations with specific content values of 17.1 nmol aromatase/mg protein. Final purified preparations showed virtually no typical P450 spectra under standard conditions, but displayed full activity with typical enzyme kinetic parameters. These unusual results suggest that standard methods of P450 measurement are inappropriate when applied to aromatase. The findings are fully consistent with those encountered previously for purified preparations from a human placental source and led us to a new aromatase quantification method based on ligand-induced difference spectroscopy. A new HPLC assay is described which rapidly separates heme and apoprotein while measuring total heme content. Matrix assisted laser desorption ionization time-of-flight mass spectrometry was employed with both glycosylated and deglycosylated forms of the final purified product to confirm its identity as a glycosylated cytochrome P450. PMID- 11483008 TI - Construction, overexpression, and purification of Arthrobacter globiformis amine oxidase-Strep-tag II fusion protein. AB - The copper-containing amine oxidase from Arthrobacter globiformis has been expressed and purified as a fusion protein with a C-terminal Strep-tag II peptide. This tag facilitates the rapid purification of the enzyme on a large scale using the StrepTactin POROS medium. For example, we have demonstrated that 50 mg of protein can be obtained in 2 days from 2 L of Escherichia coli. The purified fusion protein displays turnover and spectroscopic properties that are essentially identical to those of the wild-type enzyme. Given the location of the C-terminus in four amine oxidase crystal structures, this strategy should be quite general for the rapid purification of amine oxidases from multiple sources. PMID- 11483009 TI - Expression and purification of active recombinant ATM protein from transiently transfected mammalian cells. AB - The gene mutated in the human disease ataxia telangiectasia (AT), termed ATM, encodes a large protein kinase involved in DNA repair and cell cycle control. Biochemical characterization of ATM function has been somewhat difficult because of its large size (approximately 370 kDa) and relatively low level of expression in several systems. The majority of studies have used immunoprecipitated ATM or purified ATM obtained through relatively complex procedures. Here, we describe an efficient method for the expression and purification of FLAG-epitope-tagged recombinant human ATM protein (F-ATM). This method utilizes the expression of F ATM in transiently transfected 293T cells followed by anti-FLAG-agarose affinity chromatography. The transfection procedure has been optimized for large (225 cm(2)) culture flasks and F-ATM can be purified to near homogeneity as judged by SDS-PAGE. This procedure yields approximately 1 microg of catalytically active F ATM protein/225-cm(2) flask that can be used for biochemical studies. PMID- 11483010 TI - Cloning, expression, and purification of the Staphylococcus simulans lysostaphin using the intein-chitin-binding domain (CBD) system. AB - The Staphylococcus simulans gene encoding lysostaphin has been PCR amplified from pRG5 recombinant plasmid (ATCC 67076) and cloned into Escherichia coli expression pTYB12 vector (IMPACT-CN System, New England BioLabs) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag. The self-cleavage activity of the intein allows the release of the lysostaphin enzyme from the chitin-bound intein tag, resulting in a single-column purification of the target protein. This abundant overproduction allows purifying milligram amounts of the enzyme. PMID- 11483011 TI - A new method for protein coexpression in Escherichia coli using two incompatible plasmids. AB - It is commonly believed that incompatible plasmids carrying the same replicon cannot coexist stably in one Escherichia coli cell. However, we found that two incompatible plasmids carrying different antibiotic resistance genes, if under the selection pressure of the two antibiotics, can coexist in E. coli for at least 14 h, which is adequate for routine culture and protein expression. Based on this discovery, we developed a new method to coexpress foreign proteins in E. coli using two incompatible plasmids. The coding regions of the two subunits (DFF45 and DFF40) of the human DNA fragmentation factor (DFF) were cloned into two incompatible bacterial expression vectors-pET-21a with ampicillin resistance and pET-28a with kanamycin resistance, respectively. The two resulting plasmids were used to cotransform E. coli BL21(DE3) cells. After selection by ampicillin and kanamycin simultaneously, cotransformants that contain both recombinant plasmids were obtained. Induced by isopropyl beta-d-thiogalactoside, DFF45, and DFF40 were coexpressed efficiently in the presence of the two antibiotics. The coexpression product contained adequate soluble portions for both DFF45 and DFF40, while all DFF40 was insoluble if expressed alone. The coexpression product also exhibited the same caspase-activated DNase activity as its natural counterparts, which cannot be obtained if its two subunits are expressed separately. PMID- 11483012 TI - Expression of a VEGF-like protein from Parapoxvirus ovis in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. AB - We report on the expression of a VEGF-like protein encoded by Parapoxvirus ovis in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. We show that a lysine residue at amino acid position 2 (K2) is an important determinant for the stability of this protein in S. cerevisiae. Replacement of K2 by an arginine results in stabilization of the protein. This observation suggests that this lysine may be a target for ubiquitinylation, which is a prerequisite for proteasome-mediated protein degradation. Interestingly, in S. pombe the lysine (K2) has no influence on the stability of the protein. This result indicates that the two yeast species exhibit significant differences in their protein degradation pathways. PMID- 11483013 TI - Free polymeric bioligands in aqueous two-phase affinity extractions of microbial xylanases and pullulanase. AB - Two reversibly soluble-insoluble polymers (viz. Eudragit S-100 and alginate) were used as free macroaffinity bioligands in polyethylene glycol (PEG)/salt two-phase systems for separation of enzymes. Incorporation of Eudragit S-100 and alginate in the PEG phase led to considerable selectivity in separation of microbial xylanases and pullulanase, respectively. Xylanase from Aspergillus niger was recovered 93% with 56-fold purification, whereas the enzyme from Trichoderma reesei and Bacillus amyloliquefaciens was obtained with 93% activity recovery (31 fold purification) and 90% activity recovery (32-fold purification), respectively. From Bacillus acidopullulyticus pullulanase, 85% enzyme activity recovery with 44-fold purification was obtained. The approach described here shows the potential of developing into a general approach for use of reversibly soluble-insoluble macroaffinity ligand in two-phase affinity extraction. PMID- 11483014 TI - Large-scale preparation of recombinant ovine prolactin and determination of its in vitro and in vivo activity. AB - Recombinant bovine Ala-prolactin (PRL) (GenBank Accession No. V00112) in prokaryotic expression plasmid pMON3401 was mutated using a mutagenesis kit, to prepare plasmid encoding ovine PRL (oPRL) (GenBank Accession No. M27057) Escherichia coli cells transformed with this latter plasmid overexpressed large amounts of oPRL upon induction with nalidixic acid. The expressed protein, found in inclusion bodies, was refolded and purified to homogeneity on a Q-Sepharose column, yielding an electrophoretically pure fraction composed of over 98% monomeric protein of the expected molecular mass of approximately 23 kDa. The biological activity of the recombinant oPRL after proper renaturation was evidenced in vitro by its ability to stimulate proliferation of rat lymphoma Nb(2) cells possessing PRL receptors, to stimulate luciferase activity in HEK 293 cells transiently transfected with oPRL receptors, and to induce progesterone secretion in primary cultures of luteal cells obtained from midpregnant ewes. In contrast to ovine growth hormone or ovine placental lactogen, recombinant oPRL had no galactopoietic effect in lactating ewes. PMID- 11483015 TI - Perinatal brain injury. PMID- 11483016 TI - Molecular and biochemical mechanisms of perinatal brain injury. AB - Hypoxic-ischemic injury to the prenatal and perinatal brain is a major contributor to morbidity and mortality to infants, often leading to mental retardation, seizures, and cerebral palsy. The susceptibility of the immature CNS to hypoxia-ischemia is largely dependent on the temporal and regional status of critical developmental processes, as well as on the regulation of cerebral blood flow and metabolism. The molecular and biochemical mechanisms of acute injury to the neonatal brain in experimental rodent and murine models of hypoxic-ischemic and ischemic injury, including disturbances of intracellular homeostasis, role of glutamate receptors, free radicals and transitional ions, as well as the modifying role of gene expression to cell death/survival will be reviewed in this chapter. PMID- 11483017 TI - Hypoxic-ischemic brain injury in the newborn: pathophysiology and potential strategies for intervention. AB - There is increasing clinical and experimental data describing the evolution of hypoxic-ischemic encephalopathy in the perinatal period. Outcome to the fetus is determined not only by the impact of gross asphyxial insult, but also external factors that sensitize the brain to injury. Delayed neuronal and glial death occurring in the hours and days after the insult by apoptotic and related processes are observed following severe injury, and offer the most promise for pharmacological intervention. Furthermore, new technologies allow the identification of subtle insults with evolving encephalopathies that have implications for long-term neurological outcome. Application of this knowledge will allow us to identify strategies for early intervention and prevent the course of damage caused by hypoxic-ischemic injury. PMID- 11483018 TI - Cerebral white matter damage in the preterm infant: pathophysiology and risk factors. AB - Based on clinical, epidemiologic, and experimental studies, the aetiology of white matter damage, specifically periventricular leukomalacia (PVL), is multifactorial and involves pre- and perinatal factors possibly including genetic factors, hypoxic-ischaemic insults, infection, excess cytokines, free radical production, increased excitatory amino acid release, and trophic factor deficiencies. The article summarizes research findings about the aetiology of white matter damage and cerebral palsy in preterm infants. The information is organized according to specific antecedents, for which we present epidemiological and neurobiological data. The most important prenatal factor appears to be intrauterine infection. We discuss the evidence supporting the hypothesis that the foetal inflammatory response contributes to neonatal brain injury and later developmental disability. We recently established an animal model of excitotoxic lesions in the developing mouse brain. Brain damage was induced by intra-cortical injections of ibotenate, a glutamatergic agonist. When administered on post-natal day 5 ibotenate induced the formation of white matter cysts. Our animal model could be used to further explore the mechanisms involved in the formation of PVL. Potentially preventive strategies will be discussed. PMID- 11483019 TI - Intraventricular haemorrhage and posthaemorrhagic hydrocephalus: pathogenesis, prevention and future interventions. AB - Intraventricular haemorrhage (IVH) is still a major complication of preterm birth with serious disability resulting. The fragile blood vessels in the germinal matrix below the ventricular lining and the instability of blood flow to this highly vascular area are the main mechanisms behind IVH. There is good evidence that corticosteroid therapy before preterm delivery reduces mortality and IVH substantially with a trend towards a reduction in disability. There is good evidence that postnatal indomethacin reduces IVH but no evidence that mortality or disability is reduced. There is evidence that stabilizing cerebral blood flow with pancuronium in infants with respiratory distress reduces IVH in the minority of infants with marked fluctuations. There is limited evidence that postnatal vitamin E and ethamsylate reduce IVH but insufficient evidence of reduced mortality or disability. Hydrocephalus following IVH results initially from multiple small blood clots throughout the CSF channels impeding circulation and re-absorption. Transforming growth factor beta is released into the CSF and there is mounting evidence that this cytokine stimulates the laying down of extracellular matrix proteins such as laminin and fibronectin which produce permanent obstruction to the CSF pathways. Interventions such as early lumbar punctures, diuretic drugs to reduce CSF production and intraventricular fibrinolytic therapy have been tested and, not only fail to prevent shunt dependence, death or disability, but have significant adverse effects. Surgical interventions such as subcutaneous reservoir and external drain have not been subject to controlled trial. Ventriculoperitoneal shunt is not feasible in the early phase after IVH but, despite the problems with blockages and infections, remains the only option for infants with excessive head expansion over periods of weeks. New treatment approaches aimed at preventing hydrocephalus are needed. PMID- 11483020 TI - Hypoglycemic brain injury. AB - Hypoglycemia frequently occurs in newborn infants who previously have suffered asphyxia, who are offspring of diabetic mothers, or who are low birthweight for gestational age (IUGR). Many infants who are hypoglycemic do not exhibit clinical manifestations, while others are symptomatic and at risk for the occurrence of permanent brain damage. This review emphasizes the clinical, neuropathologic, and neuro-imaging features of hypoglycemia in newborn infants, especially those who are symptomatic. Neurologic morbidity occurs particularly in those infants who have suffered severe, protracted, or recurrent symptomatic hypoglycemia. Experimental observations emphasize the resistance of the immature brain to the damaging effect of hypoglycemia; such resistance occurs as a consequence of compensatory increases in cerebral blood flow, lower energy requirements, higher endogenous carbohydrate stores, and an ability to incorporate and consume alternative organic substrates to spare glucose for energy production. Hypoglycemia combined with hypoxia-ischemia (asphyxia) is more deleterious to the immature brain than either condition alone. PMID- 11483021 TI - Neonatal bacterial meningitis. AB - Despite major improvements in infant intensive care, neonatal meningitis remains a devastating disease. Survivors of bacterial meningitis are at high-risk for life-long neurological handicaps, and despite a reduction in mortality, the morbidity of neonatal meningitis has not changed substantially over the last thirty years. A substantial improvement in outcome is unlikely to result from further refinements in ICU technology or new antibiotics. However, recent advancements in our understanding of the pathogenesis of meningitis and the pathophysiology of brain injury in meningitis may provide the opportunity to interrupt the mechanisms that allow bacteria to enter the central nervous system and initiate the inflammatory response. Strategies aimed at modulating the inflammatory response must be chosen carefully, so as not to disrupt normal host responses needed for the infant to recover from the infectious episode. PMID- 11483022 TI - Antenatal therapies and the developing brain. AB - This chapter presents a review of basic science and human studies of two commonly used pharmacologic agents (antenatal steroids and magnesium sulfate), in pregnancies at risk of preterm delivery, and examines the effects of these therapies on the developing brain. Very low birthweight (VLBW) infants are known to be at risk of both short-term and long-term neurodevelopmental sequelae; therefore, an understanding of the mechanisms contributing to both neuroprotective and neurotoxic effects of antenatal therapies on the immature brain and potential effects on long-term outcome are critical. Although the short term beneficial effects of a single course of antenatal steroids are well documented, the experimental animal literature suggests detrimental effects on neurodevelopment of multiple doses. In addition, clinical studies of repeat doses suggest a negative impact on head and brain growth. The animal and human data on the effects of MgSO(4)are also mixed with both beneficial effects or no effects on neurodevelopment. This review will discuss the potential impact of single versus multiple doses and timing of doses on the brain. PMID- 11483023 TI - Environmental factors and disturbances of brain development. AB - Foetal and neonatal brain is under the influence of environmental factors from maternal and extra-maternal origin. Based on the available data, these environmental factors can be classified into three arbitrary groups: (i) factors and maternal status with a demonstrated deleterious effect on the foetal brain (i.e. ethanol, cocaine, some drugs including anticonvulsants, some viral infections, maternal diabetes, untreated maternal phenylketonuria); (ii) factors highly suspected to interfere with foetal brain development (i.e. lead and other heavy metals, some drugs like benzodiazepines, nicotine); (iii) factors which have been shown to be safe for the developing brain in the available studies (i.e. low to moderate doses of caffeine, methadone). However, most of these studies do not address the potential risk of environmental factors on minimal to moderate cognitive and behavioural disturbances. Finally, the impact of the neonatal environment on brain development in very pre-term infants is probably underestimated. PMID- 11483024 TI - Magnetic resonance techniques in the evaluation of the perinatal brain: recent advances and future directions. AB - Magnetic resonance (MR) techniques are attractive for use in the developing brain because of their resolving power and their relative noninvasiveness. Their ability to provide detailed structural as well as metabolic and functional information without the use of ionizing radiation is unique. Conventional MR Imaging has widely proven its potential for identifying normal and pathologic brain morphology. Functional MR imaging such as diffusion-weighted imaging (DWI) and perfusion and blood-oxygenation-dependent BOLD imaging are newer imaging methods providing insights into brain physiology. This review will focus on the application of different MR techniques including the conventional structural MR imaging techniques and the more advanced MR techniques, such as the quantitative morphometric MR methods, the diffusion weighted MR techniques, the functional MR techniques and MR spectroscopy in the study of the fetal and newborn brain. PMID- 11483025 TI - Efficient proline-catalyzed Michael additions of unmodified ketones to nitro olefins. AB - [reaction: see text] Here we describe the proline-catalyzed Michael addition of unmodified ketones to nitro olefins. This novel reaction provides gamma-nitro ketones in modest enantioselectivity yet excellent yields. PMID- 11483026 TI - Synthesis of (-)-aphanorphine using aryl radical cyclization. AB - [structure: see text] The synthesis of (-)-aphanorphine was achieved by using Bu(3)SnH-mediated aryl radical cyclization of 1-benzyloxycarbonyl-2-(2-bromo-4 methoxyphenylmethyl)-2-methoxycarbonyl-4-(phenylthiomethylene)pyrrolidine, leading to exclusive formation of the 6-exo cyclization product. PMID- 11483027 TI - Evolution of a synthetic approach to CP-263,114. AB - [structure: see text] Three different approaches to the carbocyclic core of CP 263,114 are presented that illustrate a strategic evolution from an oxy-Cope rearrangement to variants of the Wharton fragmentation. PMID- 11483028 TI - An expeditious approach toward the total synthesis of CP-263,114. AB - [reaction: see text] Assembly of the carbocyclic core of CP-263,114 has been accomplished efficiently and in high yield. Key steps include a phenolic oxidation/intramolecular Diels-Alder sequence, tandem radical cyclization, and the late-stage fragmentation of a densely functionalized isotwistane skeleton. PMID- 11483029 TI - Enhanced diastereoselectivity in asymmetric crotylation reactions using propargylic dicobalt hexacarbonyl complexes. AB - [reaction: see text] Hexacarbonyl dicobalt complexes of propargylic acetals undergo Lewis acid catalyzed crotylation reactions with enhanced levels of diastereoselectivity (dr 6 to >20:1, syn/anti) while efficiently producing stereochemically well-defined homoallylic ethers. These results are in contrast to uncomplexed propargylic acetals, which undergo the crotylation reactions with low selectivity (dr < 2:1, syn/anti). After removal of the cobalt complex, the reactions afford propargylic ethers in high yields. PMID- 11483030 TI - Solid-state supramolecular structures of resorcinol-arylboronic acid compounds. AB - [structure: see text] An X-ray crystallographic study of unique hydrogen-bonded supramolecular solid-state networks comprised of a tetraarylboronic acid resorcinarene is described. When 1 is recrystallized from 9:1 MeOH:EtOH, partial esterification takes place to give compound 2, the corresponding half methyl ester, which forms an infinite two-dimensional array. Each molecule participates in 12 hydrogen bonds with other macrocycles. These hydrogen bonds are both B-OH- - OH (phenolic) and OH (phenolic)- - -OH (phenolic). PMID- 11483031 TI - One-step transformation of tricyclopentabenzene (trindane, C(15)H(18)) to bicyclo(10.3.0)pentadec-1(12)ene- 2,6,7,11-tetrone (C(15)H(18)O(4)) and its aldol product, 12-hydroxy-16-oxatetracyclo(10.3.1.0.(1,5)0(7,11))hexadec-7(11)ene-2,6 dione (C(15)H(18)O(4)). AB - [reaction: see text] Ozonolysis of 1 largely results in 2 and 3, having features similar to several classes of natural products. The retention of the C(15) pericycle suggests preference for the cleavage of pi-bonds endo to the cyclopentane ring. This unique property of trindane offers opportunities for synthesis of complex natural products from this hydrocarbon that can be made in quantity by acid-catalyzed trimerization of cyclopentanone. PMID- 11483032 TI - Synthesis of the diazonamide A macrocyclic core via a Dieckmann-type cyclization. AB - [reaction: see text] Suzuki coupling of 18 and 30 affords 9 as an interconverting mixture of two atropisomers. Treatment with LDA at -23 degrees C affords the macrocyclic ketone 8 in 57% yield. PMID- 11483033 TI - The first 2,2-dialkoxythiirane. AB - [reaction: see text] Thermolysis of 1 at 110 degrees C in benzene containing adamantanethione leads to thiirane 2 in 92% yield, as an isolable, stable solid. Compound 2 is the first example of the hitherto unknown 2,2-dialkoxythiiranes. It shows some reactions characteristic of thiiranes. PMID- 11483034 TI - Contribution of a pyrene fluorescence probe to the aggregation propensity of polypeptides. AB - [structure: see text] Two synthetic polypeptides, TT1p and TT1b, have been used in comparative aggregation equilibrium studies. The findings reveal that a single alkylpyrene moiety in TT1p contributes about 30% of the polypeptide dimerization energy in aqueous media. This result not only is informative with regard to the aggregation properties of these particular photoactive polypeptides but also provides a quantitative understanding of the limitations on the use of pyrene chromophores as emission probes. PMID- 11483035 TI - Synthesis and biological evaluation of novel macrocyclic paclitaxel analogues. AB - [reaction: see text] This work describes the synthesis of two novel macrocyclic taxoid constructs by ring-closing olefin metathesis (RCM) and their biological evaluation. Computational studies examine conformational profiles of 1 and 2 for their fit to the beta-tubulin binding site determined by electron crystallography. The results support the hypothesis that paclitaxel binds to microtubules in a "T" conformation. PMID- 11483036 TI - Dissected nucleus-independent chemical shift analysis of pi-aromaticity and antiaromaticity. AB - [structure: see text] Analysis of the basic pi-aromatic (benzene) and antiaromatic (cyclobutadiene) systems by dissected nucleus-independent chemical shifts (NICS) shows the contrasting diatropic and paratropic effects, but also reveals subtleties and unexpected details. PMID- 11483037 TI - Novel fluorophores: efficient synthesis and photophysical study. AB - [structure: see text] We have synthesized novel fluorophores by using Sonogashira reactions of 1,4-bis(dibromovinyl)benzene and 2,5-bis(dibromovinyl)thiophene with various aromatic bromides. The emission maxima of these fluorophores vary from the indigo blue to the reddish-orange region, depending on the structures of aromatic nuclei and peripheral moieties. PMID- 11483038 TI - Versatile strategy for oligonucleotide derivatization. Introduction of lanthanide(III) chelates to oligonucleotides. AB - [reaction: see text] Novel nucleosidic phosphoramidite blocks were synthesized by a Mitsunobu reaction between 2'-deoxy-5'-O-(4,4'-dimethoxytrityl)uridine and a primary alcohol containing a conjugate group in its structure (a protected functional group, an organic dye, or a precursor of a lanthanide(III) chelate) followed by phosphitylation. They were used in machine-assisted DNA synthesis in the standard manner. A slightly modified deprotection procedure was used for the preparation of oligonucleotide conjugates tethered to lanthanide(III) chelates. For the latter application one non-nucleosidic block was also synthesized. PMID- 11483039 TI - Mild and selective sodium azide mediated cleavage of p-nitrobenzoic esters. AB - [reaction: see text] A mild and selective cleavage of p-nitrobenzoic esters by sodium azide in methanol is reported. This new methodology is mild enough for use with acid- or base-sensitive compounds. No elimination byproducts are formed. Fmoc- and trifluoroacetyl-amino protecting groups, benzyl esters, and ethyl esters remain unaffected. Less reactive compounds are discussed in terms of steric factors, and yields are increased by altering the azide solvation. PMID- 11483040 TI - Short, highly efficient syntheses of protected 3-azido- and 4-azidoproline and their precursors. AB - [structure: see text] An improved synthesis of protected cis- and trans-3-azido-L proline and cis- and trans-4-azido-L- and -D-proline is reported. These compounds have been synthesized from the corresponding hydroxyproline precursors using diphenylphosphoryl azide under Mitsunobu conditions. Short, highly efficient syntheses of these precursors are described, based on a new lactone-opening reaction and p-nitrobenzoate hydrolysis under very mild conditions. PMID- 11483041 TI - Facile synthesis of rotaxanes through condensation reactions of DCC-[2]rotaxanes. AB - [reaction: see text] In this Letter, we present an easy method for the synthesis of rotaxanes using a novel DCC-[2]rotaxane. The DCC-[2]rotaxane is composed of dibenzo-24-crown-8 ether, an amino acid tether, and di-tert-butyl phenyl rings as blocking groups. It is relatively stable and can be purified by column chromatography. A series of model rotaxanes were obtained in good yields by condensing the DCC-[2]rotaxanes with N-(2-aminoethyl)-3,5-di-tert butylbenzylamide in acetonitrile and chloroform. PMID- 11483042 TI - Regiochemical control in intramolecular cyclization of methylene-interrupted epoxydiols. AB - [reaction: see text] Methylene-interrupted epoxydiols have multiple regiochemical routes for cyclization. The 5-exo process is the most prevalent under acidic conditions. However, the regioselectivity can be controlled by the appropriate choice of acid promoter and pendant groups adjacent to the epoxide. The 5-exo product is obtained exclusively without the presence of a carbocation-stabilizing pendant group. Alkenyl and thiophenyl groups adjacent to the epoxide alter the regioselectivity and enable access to the 5-endo tetrahydrofuran and 6-endo tetrahydropyran products. PMID- 11483043 TI - Heterogenization of a chiral bis(oxazoline) catalyst by grafting onto silica. AB - [reaction: see text] We report here the first heterogenization of a bis(oxazoline) ligand on an inorganic (silica) surface. The activity and enantioselectivity of this new material as a catalyst for the Diels-Alder reaction were checked, and it was shown that under certain conditions enantioselectivities similar to those of the homogeneous catalyst are reached. It was also shown that under these conditions the catalyst can be recycled without loss of activity or selectivity. PMID- 11483044 TI - Direct asymmetric aldol reactions of acetone using bimetallic zinc catalysts. AB - [reaction: see text] The enantioselective aldol reaction using a novel binuclear zinc catalyst of acetone with several aldehydes gave products in good yields (62 89%) with a high level of enantioselectivity (ee = 76-92%). PMID- 11483045 TI - Preparation of 2,5-disubstituted oxazoles from N-propargylamides. AB - [reaction: see text] 2,5-Disubstituted oxazoles have been prepared through the reaction of N-propargylamides with aryl iodides in the presence of Pd(2)(dba)(3), tri(2-furyl)phosphine, and NaO(t)()Bu. The reaction appears to proceed through a palladium-catalyzed coupling step followed by the in situ cyclization of the resultant coupling product. PMID- 11483046 TI - First total synthesis of (+/-)-stemonamide and (+/-)-isostemonamide. AB - [structure: see text] The total synthesis of the tetracyclic alkaloids stemonamide (1) and isostemonamide (2) is presented. The key step is the reaction between a silyloxyfuran and an N-acyliminium ion. The second quaternary center is created by an intramolecular aldol spirocyclization. After 1,4-addition of an appropriate side chain, the methyl and double bond are installed by Mannich reaction. The seven-membered ring is closed by intramolecular nucleophilic displacement. PMID- 11483047 TI - Chemoselective Pd(0)-catalyzed peptide coupling in water. AB - [reaction: see text] Highly charged peptides ranging in length from 17 to 33 residues have been efficiently tricoupled to a trialkyne nucleus by using a Sonogashira Pd(0) coupling strategy under both acidic (pH 5.0) and basic (pH 7.5) conditions. These results demonstrate the utility of Pd(0) to construct protein sized structures (12,000 mol wt) in aqueous milieu. PMID- 11483048 TI - Stereoselective synthesis of styrene oxides via a Mitsunobu cyclodehydration. AB - [reaction: see text] The Mitsunobu cyclodehydration of chiral phenethane-1,2 diols (4), readily accessed from the styrene derivative (5), has been demonstrated to provide the corresponding styrene oxides (2) with high levels of stereoretention (up to 99%). Optimized reaction conditions are described, from which the combination of tricyclohexylphosphine (Chx(3)P) and diisopropylazodicarboxylate (DIAD) in THF and R = EWG provides the best results. PMID- 11483049 TI - Improved chemical synthesis of UDP-galactofuranose. AB - [reaction: see text] A reliable and efficient synthetic route to UDP-alpha-D galactofuranose (UDP-Galf) has been developed. Reaction of UMP-N methylimidazolide with Galf 1-phosphate proceeds rapidly to provide UDP-Galf with excellent reproducibility and in a yield approximately twice as high as those reported previously. PMID- 11483050 TI - Remarkable rate acceleration of Pd(0)-catalyzed hydrogermylation of alkynes and dienes in water. AB - [reaction: see text] Without the use of hydrophilic cosolvents or phase transfer catalysts, Pd(0)-catalyzed hydrogermylation of alkynes in water provides dienylgermanes efficiently with high stereo- and regioselectivity. The reaction in water proceeds much faster than the reaction under neat conditions. By using an aqueous system, hydrogermylation can be conducted at a low catalyst loading. PMID- 11483051 TI - Synthesis and application of chiral P,N-ligands with pseudo-meso and pseudo-C(2) symmetry. AB - [structure: see text] Two diastereoisomeric P,N-ligands, (S,S)-1 and (R,S)-1, were synthesized and assessed in palladium-catalyzed allylic alkylations. With rac-1,3-diphenylpropenyl acetate as substrate, ligand (S,S)-1, with "pseudo-C(2)" symmetry, exhibited higher reactivity and higher enantioselectivity than the "pseudo-meso" ligand (R,S)-1, whereas reversal reactivity and selectivity were observed with rac-3-cyclohexenyl acetate. PMID- 11483052 TI - Novel [1,2]- and [2,3]-Wittig rearrangements of alpha-benzyloxy beta-CF(3)-beta lactam enolates. AB - [reaction: see text] alpha-Benzyloxy alpha-CF(3)-beta-lactams are shown to offer the first examples of the enolate [1,2]- and enolate ortho-[2,3]-Wittig rearrangements which provide a unique entry to the alpha-benzyl-alpha-hydroxy lactams and the alpha-aryl-alpha-hydroxy lactams, respectively. Both products are potential precursors of new trifluoromethyl isoserines, and the latter is not accessible via the usual alkylation methodology. PMID- 11483053 TI - A 4 + 3 cycloaddition approach to the synthesis of spatol. A formal total synthesis of racemic spatol. AB - [reaction: see text] A formal total synthesis of racemic spatol is presented. The key steps involved a 4 + 3 cycloaddition of a halogenated cyclopentenyl cation to cyclopentadiene and a quasi-Favorskii rearrangement. PMID- 11483054 TI - Gold(III) chloride catalyzed cyclization of alpha-hydroxyallenes to 2,5 dihydrofurans. AB - [reaction: see text] Functionalized alpha-hydroxyallenes 1 were smoothly converted into the corresponding 2,5-dihydrofurans 2 by using 5-10 mol % of gold(III) chloride as catalyst. This mild and efficient cyclization method can be applied to alkyl- and alkenyl-substituted allenes at room temperature, furnishing tri- and tetrasubstituted dihydrofurans in good to excellent chemical yields and with complete axis to center chirality transfer. PMID- 11483055 TI - 3-Aryl-2-oxazolidinones through the palladium-catalyzed N-arylation of 2 oxazolidinones. AB - [reaction: see text] 3-Aryl-2-oxazolidinones are obtained in good yields through the palladium-catalyzed N-arylation of 2-oxazolidinones with aryl bromides. The nature of aryl bromides, phosphine ligands, bases, and solvents strongly affects the reaction outcome. PMID- 11483056 TI - Reductive desymmetrization of 2-alkyl-1,3-diketones catalyzed by optically active beta-ketoiminato cobalt complexes. AB - [reaction: see text] The reductive desymmetrization of acyclic 1,3-diketones was achieved for the first time by catalytic borohydride reduction in the presence of optically active beta-ketoiminato cobalt(II) complex catalysts. In this reaction, various 2-substituted-1,3-diaryl-1,3-propanediones were converted into the corresponding optically active 2-substituted-1,3-diaryl-3-hydroxypropanone in good-to-high yields with excellent diastereo- and enantioselectivities and high catalytic efficiencies. PMID- 11483057 TI - Synthesis of ring B unsaturated estriols. Confirming the structure of a diagnostic analyte for Smith-Lemli-Opitz syndrome. AB - [structure: see text] Brief partial syntheses are described for ring B unsaturated estriols, which are candidate metabolites diagnostic for Smith-Lemli Opitz syndrome prenatally. These steroids are also likely metabolites of the Premarin preparation used in estrogen replacement therapy. Equilin (8) was converted in three steps to 7-dehydroestriol, which was isomerized to 8 dehydroestriol. The simplicity of the transformations belies the lability of these previously inaccessible metabolites and their synthetic precursors. PMID- 11483058 TI - Immobilization of chiral ligands on polymer fibers by electron beam induced grafting and applications in enantioselective catalysis. AB - [reaction: see text] Styrenic TADDOL and L-prolinol-derived monomers were immobilized on polyethylene fibers by electron beam induced preirradiation grafting using styrene as comonomer. The polymer-supported chiral ligands were utilized as catalysts in the asymmetric addition of diethylzinc to benzaldehyde. Fiber-bound titanium TADDOLate gave a quantitative conversion of benzaldehyde to 1-phenylpropan-1-ol in a 97:3 S/R enantiomeric ratio. The catalyst was successfully regenerated and employed in subsequent reactions with retention of high enantioselectivities. PMID- 11483059 TI - Novel approach for the stereocontrolled construction of eudesmane skeleton: a concise synthesis of (+/-)-balanitol. AB - [structure: see text] A novel method for the stereocontrolled construction of the eudesmane ring system based on a cationic cyclization is presented, and the approach is exemplified in a short and efficient total synthesis of (+/-) balanitol (2). PMID- 11483060 TI - Evaluation of hydrogen bonding complementarity between a secondary sulfonamide and an alpha-amino acid residue. AB - [structure: see text] We report an initial step toward the development of sulfonamide-based complements for extended peptide strands. A molecule containing one secondary sulfonamide unit and one valine residue linked by a turn-forming segment was found by IR and NMR to exhibit a doubly hydrogen-bonded folding pattern in chloroform. PMID- 11483061 TI - Intramolecular cyclopropanation of glycals: studies toward the synthesis of canadensolide, sporothriolide, and xylobovide. AB - [reaction: see text] The first examples of copper-catalyzed intramolecular cyclopropanations of glycal-derived diazoacetates are reported. The new cyclopropanes are converted into advanced intermediates for the synthesis of bislactone natural products. Synthetic highlights include the selective monodeprotection of a di-tert-butylsilylene ether and a zinc-mediated ring opening cascade reaction. PMID- 11483062 TI - Synthesis of stereotriads by oxymercuration of substituted cyclopropylcarbinols. AB - [structure: see text] Cyclopropylcarbinol derivatives bearing an adjacent methyl substituted stereocenter and a remote beta-hydroxy group protected as a pivalate underwent anchimerically assisted regio- and diastereoselective oxymercurations, affording after reductive demercuration, an access to stereotriads. PMID- 11483063 TI - Stereoselective C-glycoside formation by a rhodium(I)-catalyzed 1,4-addition of arylboronic acids to acetylated enones derived from glycals. AB - [reaction: see text] A new method for the formation of C-glycosides has been developed employing a cationic rhodium(I)-catalyzed 1,4-addition of arylboronic acids to enones derived from glycals. The reaction is stereoselective for the alpha-anomer and is highly dependent on the nature of the rhodium catalyst. PMID- 11483064 TI - Intramolecular 1,3-dipolar cycloaddition strategy for enantioselective synthesis of FR-900482 analogues. AB - [reaction: see text] Enantioselective synthesis of FR-900482 analogues is described. The key reaction of the synthesis is intramolecular 1,3-dipolar cycloaddition of a highly functionalized nitrile oxide with complete stereo- and regioselectivities to construct the eight-membered benzazocine ring. PMID- 11483065 TI - Ruthenium complex-catalyzed direct ortho arylation and alkenylation of 2 arylpyridines with organic halides. AB - [reaction: see text] The ortho position of the aromatic ring of pyridyl group substituted aromatic compounds is directly arylated or alkenylated with organic halides in the presence of a catalytic amount of a ruthenium(II)-phosphine complex. PMID- 11483066 TI - CuI-catalyzed coupling reaction of beta-amino acids or esters with aryl halides at temperature lower than that employed in the normal Ullmann reaction. Facile synthesis of SB-214857. AB - [reaction: see text] The CuI-catalyzed coupling reaction of aryl halides with beta-amino acids or beta-amino esters is completed at 100 degrees C in 48 h, which indicates that the structure of the beta-amino acid has an accelerating effect for the Ullmann-type aryl amination reaction. This coupling reaction can be used to prepare enantiopure N-aryl beta-amino acids. An efficient synthetic route to SB214857, a potent GPIIb/IIIa receptor antagonist, is developed using this method. PMID- 11483067 TI - Asymmetric epoxidation of olefins catalyzed by chiral iminium salts generated in situ from amines and aldehydes. AB - [reaction: see text] A new approach for catalytic asymmetric epoxidation of olefins was developed that utilized chiral iminium salts, generated in situ from chiral amines and aldehydes, as catalysts. Epoxidation reactions can be conducted with 20 mol % of amines and aldehydes. The enantioselectivity of epoxides can be up to 65%. This modular approach obviates the difficulties inherent in the preparation and isolation of unstable exocyclic iminium salts. PMID- 11483068 TI - Synthetic utility of stannyl enolates as radical alkylating agents. AB - [reaction: see text] The radical-initiated beta-ketoalkylation of haloalkanes with tributylstannyl enolates is described. Stannyl enolates derived from aromatic ketones are reactive toward the homolytic beta-ketoalkylation of simple haloalkanes as well as those activated by an electron-withdrawing group. The reactivity of stannyl enolates as radical alkylating agents can be utilized for an efficient three-component coupling reaction among stannyl enolates, haloalkanes, and electron-deficient alkenes. PMID- 11483069 TI - A novel approach toward low optical band gap polysquaraines. AB - [reaction: see text] Polycondensation of 2,5-dialkoxydivinylbenzene-bridged bispyrroles and squaric acid resulted in extensively conjugated polymers with strong near-infrared (NIR) absorption around 800-1000 nm, which is a signature of their low band gap (E(g)). Conjugated polymers with such a strong NIR absorption are rare. One of the synthesized polysquaraines showed a significantly low E(g) of 0.79 eV with an intrinsic conductivity of 5.3 x 10(-4) S/cm. PMID- 11483070 TI - Why is alkylation of an enolate accompanied by so much polyalkylation? AB - [reaction: see text] The lithium enolate 1-Li of 6-phenyl-alpha-tetralone forms a monomer-tetramer equilibrium in THF at 25 degrees C with K(1,4) = 4.7E+10 M(-3). The lithium enolate 2-Li, however, forms a monomer-dimer equilibrium with K(1,2) = 3800 M(-1). In both cases reaction with benzyl bromide is dominantly with the monomer. The results support an earlier conjecture of House that alkylation of an enolate is frequently accompanied by extensive polyalkylation because the less substituted enolates are more aggregated. PMID- 11483071 TI - Indium metal as a reducing agent. Selective reduction of the carbon-carbon double bond in highly activated conjugated alkenes. AB - [reaction: see text] Indium metal in aqueous ethanolic ammonium chloride reduces the C=C bond in highly activated conjugated alkenes such as alpha,alpha-dicyano olefins, beta-arylenones, and enone esters. PMID- 11483072 TI - Synthesis of bridged medium-sized rings through the intramolecular Pauson-Khand reaction. AB - [reaction: see text] A series of aromatic enynes containing steric buttressing elements were prepared and evaluated in the NMO-mediated Pauson-Khand cyclization. O-Allyl systems led to the expected angularly fused products, whereas the O-butenyl and O-pentenyl derivatives afforded the unprecedented bridge systems. PMID- 11483073 TI - Total synthesis of the sesquiterpene (+/-)-illudin C via an intramolecular nitrile oxide cycloaddition. AB - [reaction: see text] A convergent total synthesis of illudin C is described. The tricyclic ring system of the natural product was quickly assembled from cyclopropane and cyclopentene precursors via a novel oxime dianion coupling reaction and a subsequent intramolecular nitrile oxide-olefin cycloaddition. PMID- 11483074 TI - Chloroprene as a source of fine chemicals: palladium-catalyzed synthesis of terminal allenes. AB - [reaction: see text] Several functionalized terminal allenes were prepared in moderate to excellent yield by a palladium-catalyzed reaction of chloroprene (2 chloro-1,3-butadiene) with soft nucleophiles. PMID- 11483076 TI - Multicenter Trial of Cryotherapy for Retinopathy of Prematurity: ophthalmological outcomes at 10 years. AB - OBJECTIVE: To evaluate outcomes at 10 years after randomization for eyes undergoing cryotherapy vs eyes serving as controls, for patients enrolled in the Multicenter Trial of Cryotherapy for Retinopathy of Prematurity (CRYO-ROP). METHODS: The randomized cohort originally consisted of 291 preterm children with birth weights less than 1251 g who developed a defined threshold of ROP severity in one or both eyes. Patients with bilateral threshold ROP (n = 240) were randomly assigned to receive cryotherapy to one eye and no cryotherapy to the other eye. Those with ROP of less severity than threshold in the fellow eye ("asymmetric"; n = 51) were randomly assigned to cryotherapy or no cryotherapy in the eye with threshold ROP. Ten years later, a tester who was masked to treatment status of each eye measured distance and near visual acuity, with "unfavorable" outcome being 20/200 or worse. Patients also were evaluated by study-certified ophthalmologists who assessed ROP residua primarily in the posterior pole of the fundus, with unfavorable outcome being a posterior retinal fold or worse. RESULTS: For the 247 children examined, both functional and structural primary outcomes showed fewer unfavorable outcomes in treated vs control eyes: 44.4% vs 62.1% (P<.001) for distance visual acuity and 27.2% vs 47.9% (P<.001) for fundus status. Near acuity results were similar to those for distance (42.5% vs 61.6%; P<.001). Total retinal detachments had continued to occur in control eyes, increasing from 38.6% at 5(1/2) years to 41.4% at 10 years, while treated eyes remained stable (at 22.0%). A previously disturbing subgroup trend that more control eyes than treated eyes had visual acuity of 20/40 or better (in the 5(1/2)-year report) was no longer present at 10 years; eyes that received cryotherapy were found at least as likely as control eyes to have 20/40 or better visual acuity. CONCLUSIONS: At 10 years, eyes that had received cryotherapy were much less likely than control eyes to be blind. A previous trend for a higher proportion of sighted control eyes than sighted treated eyes to show acuity in the normal range was not confirmed. The results show long-term value from cryotherapy in preserving visual acuity in eyes with threshold ROP. PMID- 11483077 TI - Effect of retinal ablative therapy for threshold retinopathy of prematurity: results of Goldmann perimetry at the age of 10 years. AB - OBJECTIVE: To examine monocular visual fields at the age of 10 years in children with birth weights less than 1251 g in whom severe acute-phase retinopathy of prematurity (ROP) developed in 1 or both eyes and who had random assignment of eyes to cryotherapy (treated) or no cryotherapy (control) and in a comparison group of children who did not develop ROP in the neonatal period. METHODS: Subjects were 255 children who developed severe ROP and 104 children who did not develop ROP. All were born between January 1, 1986, and November 30, 1987, and had birth weights of less than 1251 g. Goldmann perimetry was used to measure visual field extent along 8 meridia (15 degrees, 60 degrees, 105 degrees, 150 degrees, 195 degrees, 240 degrees, 285 degrees, and 330 degrees ) using the V4e and III4e stimuli. Data were analyzed by quadrant and overall solid angle of the area of the retina sensitive to the test stimulus. RESULTS: When blind eyes were assigned a score of 0, visual field area was 24% to 26% larger in treated eyes than in control eyes. When data from only patients with 1 sighted treated eye and 1 sighted control eye were examined, visual field area was 5% smaller in treated eyes than in control eyes. For these patients, visual field area was reduced by 30% to 37% in treated eyes and by 27% to 33% in control eyes compared with eyes with no ROP. CONCLUSIONS: Cryotherapy preserves peripheral vision in eyes with severe ROP by preserving sight in these eyes. After excluding 70 blind treated and 102 blind control eyes, data from eyes with quantifiable visual fields indicate that cryotherapy produces a small reduction of visual field area in eyes with severe ROP. With or without cryotherapy, visual field area is considerably smaller in eyes that had severe acute-phase ROP than in eyes of preterm children who did not develop ROP. PMID- 11483078 TI - Contrast sensitivity at age 10 years in children who had threshold retinopathy of prematurity. AB - OBJECTIVE: To evaluate monocular contrast sensitivity (CS) at age 10 years in eyes that underwent cryotherapy and in control eyes of participants in the randomized trial of cryotherapy for retinopathy of prematurity (CRYO-ROP), and in a comparison group of 10-year-old low-birth-weight children who did not develop ROP in the neonatal period. METHODS: Eligible participants were the 255 survivors of the group of 291 infants with severe (threshold) ROP in 1 or both eyes who were enrolled in the randomized CRYO-ROP trial as neonates, as well as 104 children in the CRYO-ROP study who did not develop ROP. All had birth weights less than 1251 g. Contrast sensitivity was measured using Pelli-Robson charts at a test distance of 1 m with luminance at or higher than 64 candelas (cd)/m(2). Contrast sensitivity was estimated using the total number of letters identified. RESULTS: Unfavorable CS outcomes (<27 letters identified) were found in a greater proportion of control eyes (59.3%) than of treated eyes (39.7%) (P<.001). In patients with bilateral threshold ROP who had quantifiable CS in both eyes, CS results were similar in treated and control eyes, suggesting no detrimental effect of cryotherapy. In the "no ROP" group, 96.9% of eyes showed CS in the normal range, compared with 48.1% of treated eyes and 34.6% of control eyes. CONCLUSIONS: The results further confirm the beneficial effect of cryotherapy on visual function, and show no evidence of adverse effects of cryotherapy on CS. With or without cryotherapy, eyes with severe ROP showed substantially poorer CS than did eyes of preterm children who did not develop ROP. PMID- 11483079 TI - Reproducibility of retinal mapping using optical coherence tomography. AB - OBJECTIVE: To assess the reproducibility of retinal thickness measurement using commercially available mapping software of optical coherence tomography (OCT). METHODS: Six radial scans, 6 mm long and centered on the fixation point, were performed on 10 eyes of 10 healthy volunteers and 10 eyes of 10 diabetic patients with clinically significant macular edema. Retinal thickness was measured automatically using the mapping software of OCT in the 9 macular Early Treatment Diabetic Retinopathy Study areas and in a central area 500 microm in diameter. Measurement reproducibility was tested by means of 3 series of scans performed by 2 different observers on 2 different days. Results were assessed by their repeatability and intraclass correlation coefficients (ICCs). RESULTS: In healthy subjects, intraobserver, interobserver, and intervisit reproducibility of retinal thickness measurements were excellent, with a repeatability coefficient of less than 7 microm and ICCs of greater than 0.89. In diabetic patients, the repeatability coefficient was less than 21 microm in all areas of the macula except one, with an ICC of greater than 0.98. Relative variations in measurements were small in both healthy and diabetic subjects, with reproducibilities of +/- 5% and +/- 6%, respectively. CONCLUSION: Retinal mapping software of OCT allows reproducible measurement of retinal thickness in both healthy subjects and diabetic patients with macular edema. PMID- 11483080 TI - Age-related maculopathy in a randomized trial of low-dose aspirin among US physicians. AB - OBJECTIVE: To examine the development of age-related maculopathy (ARM) in a large scale trial of low-dose aspirin treatment. METHODS: The Physicians' Health Study I was a randomized, double-masked, placebo-controlled trial of low-dose aspirin (325 mg every other day) and beta carotene (50 mg every other day) in the prevention of cardiovascular disease and cancer conducted among 22 071 US male physicians aged 40 to 84 years in 1982. A total of 21 216 participants did not report ARM at baseline, were followed up for at least 7 years, and are included in this analysis. MAIN OUTCOME MEASURES: Total ARM, defined as a self-report confirmed by medical record evidence of an initial diagnosis subsequent to randomization, and ARM with vision loss, defined as total ARM but with vision loss to 20/30 or worse attributable to ARM. RESULTS: Early termination of the randomized aspirin component of the Physicians' Health Study I, after an average of 60.2 months of treatment and follow-up due to a statistically extreme 44% reduced risk of first myocardial infarction, resulted in a far lower number of incident cases of ARM during the aspirin treatment period than would have accrued without early termination. Thus, during an average of 60.2 months of follow-up, a total of 117 cases of ARM were confirmed, including 57 cases responsible for vision loss to 20/30 or worse. There were 51 cases of ARM in the aspirin group and 66 in the placebo group (relative risk, 0.77; 95% confidence interval, 0.54 1.11). For ARM with vision loss, there were 25 cases in the aspirin group and 32 in the placebo group (relative risk, 0.78; 95% confidence interval, 0.46-1.32). CONCLUSIONS: These randomized trial data tend to exclude any large beneficial effect of 5 years of low-dose aspirin treatment on ARM. However, a smaller, but potentially important, beneficial effect cannot be ruled out and would require testing in randomized trials of adequate size and duration. PMID- 11483081 TI - Fifteen-year outcome of surgery for the near angle in patients with accommodative esotropia and a high accommodative convergence to accommodation ratio. AB - OBJECTIVE: To determine the 15-year outcome of patients with partly accommodative esotropia with a high accommodative convergence to accommodation (AC/A) ratio who underwent surgery based on the angle of esotropia at one-third meter while wearing full hyperopic correction. METHODS: A retrospective chart review to determine the 15-year outcome of 25 patients whose 6-month outcome had been previously reported as part of a prospective, randomized, masked clinical trial. All patients had partly accommodative esotropia with a high AC/A ratio and underwent surgery based on their esotropia at one-third meter while wearing full distance optical correction. RESULTS: Fifteen years after surgery, 19 of the 22 patients for whom follow-up data are available had between 0 and less than 10 prism diopters of esotropia. Only 6 of the 19 needed to continue to wear optical correction to maintain satisfactory alignment; however, 8 more needed spectacles for visual purposes. Only 1 patient needed to use a bifocal add to have satisfactory alignment at one-third meter. All patients showed some degree of sensory fusion, with 4 obtaining 40 seconds of stereopsis and another 8 obtaining between 60 and 200 seconds of stereopsis. CONCLUSION: Surgery for the near angle obtained with patients wearing their full hyperopic distance correction provides excellent motor and sensory results in patients with partly accommodative esotropia with a high AC/A ratio. PMID- 11483082 TI - Effect of chromatic aberration on contrast sensitivity in pseudophakic eyes. AB - OBJECTIVE: To evaluate the effect of chromatic aberrations in pseudophakic eyes with various types of intraocular lenses (IOLs). PATIENTS AND METHODS: The study included 51 eyes of 33 patients who underwent cataract surgery. The eyes were divided into 3 groups according to the material from which their IOL was made: group 1, polymethyl methacrylate; group 2, silicone; and group 3, an acrylate/methacrylate copolymer. Ten normal phakic control eyes (group 4) underwent the same examination. Best-corrected distance visual acuity and contrast sensitivity were measured under white light and monochromatic light with wavelengths of 470 nm, 549 nm, and 630 nm, with the best correction under white light. RESULTS: There were no significant differences in best-corrected visual acuity and contrast sensitivity under the 549-nm monochromatic light in any group. However, under both white multichromatic light and 470- and 630-nm monochromatic light, the mean contrast sensitivity in group 3 tended to be lower, sometimes significantly, than in the other IOL groups. CONCLUSIONS: Our results showed that longitudinal chromatic aberrations of some IOLs may degrade the quality of the retinal image. Attention must be paid to the detailed optical performance of IOL materials to achieve good visual function. PMID- 11483083 TI - Effect of larger ablation zone and transition zone on corneal optical aberrations after photorefractive keratectomy. AB - OBJECTIVE: To evaluate the effects of photorefractive keratectomy on corneal optical aberrations using a 5.5-mm optical zone and a 7.0-mm transition zone. METHODS: Videokeratographs of 72 eyes from 47 patients treated for low to moderate (1-9 diopters) myopia were obtained at the preoperative and 1-, 3-, 6-, 12-, and 18-month postoperative examinations. The videokeratoscopy data files were used to calculate the wavefront variance of the corneas for small (3-mm) and large (7-mm) pupils using a previously described method. RESULTS: In general, all optical aberrations decreased postoperatively for 3-mm pupils and increased for 7 mm pupils compared with preoperative values. For 3-mm pupils, the 2 common optical aberrations (comalike [S(3)] and spherical-like [S(4)]) decreased postoperatively and never returned to preoperative values. For 7-mm pupils, however, comalike aberrations increased slightly and spherical-like aberrations increased by nearly an order of magnitude during the postoperative period. Similarly, for 3-mm pupils, the higher order S(5) and S(6) aberrations decreased throughout the postoperative period, with S(6) values showing an approximately 23 fold reduction at 12 and 18 months. For 7-mm pupils, S(5) and S(6) aberrations increased slightly, more so for S(5) (approximately 3-fold) than for S(6). Total wavefront aberrations decreased an average of 2.3 times postoperatively for 3-mm pupils, and increased significantly (P<.05) at all postoperative examinations for 7-mm pupils. Opening the pupil from 3 mm to 7 mm before surgery produced a 14 fold increase in total aberrations, whereas this same change produced an average 113-fold increase after photorefractive keratectomy. CONCLUSIONS: Corneal optical aberrations after photorefractive keratectomy with a larger ablation zone and a transition zone are less pronounced and more physiologic than those associated with first-generation (5-mm) ablations with no transition zone. CLINICAL RELEVANCE: Evaluating the postoperative corneal aberration structure will help us devise ways to minimize the wavefront aberrations of the eye through the creation of an ideal corneal first surface, thereby improving visual results for patients undergoing excimer laser ablations for refractive correction. PMID- 11483084 TI - Increased matrix metalloproteinases 1, 2, and 3 in the monkey uveoscleral outflow pathway after topical prostaglandin F(2 alpha)-isopropyl ester treatment. AB - OBJECTIVE: To investigate the effects of topical prostaglandin F(2 alpha)- isopropyl ester (PGF(2 alpha)-IE) administration on immunoreactivity of matrix metalloproteinases (MMPs) 1, 2, and 3 within the anterior segment tissues of monkey eyes. METHODS: Eight eyes from 4 cynomolgus monkeys were evaluated. One eye from each monkey was treated with 2 mg of PGF(2 alpha)-IE twice daily for 5 days, and intraocular pressure reduction was measured. After fixation and processing, deparaffinized sections of anterior segments were immunostained using antibodies to MMP-1 (interstitial collagenase), MMP-2 (gelatinase A), or MMP-3 (stromelysin-1). Optical density along 2 line segments overlying the iris root, ciliary muscle, and adjacent sclera and perpendicular to their long axes was measured using imaging densitometry. RESULTS: Compared with the contralateral vehicle-treated eyes, statistically significant increases in optical density scores were observed in the iris root, ciliary muscle, and adjacent sclera for all 3 MMPs (P<.01). In these tissues, MMP-1 immunoreactivity was increased by a mean +/- SD of 89% +/- 16%, 61% +/- 8%, and 66% +/- 57%, respectively; MMP-2 immunoreactivity by 129% +/- 53%, 82% +/- 27%, and 267% +/- 210%, respectively; and MMP-3 immunoreactivity by 207% +/- 84%, 83% +/- 49%, and 726% +/- 500%, respectively. CONCLUSIONS: Treatment of monkey eyes with PGF(2 alpha)-IE induces elevation of MMP-1, MMP-2, and MMP-3 in tissues of the uveoscleral outflow pathway. These increases suggest that MMPs might play an important role in the increased uveoscleral outflow observed with topical prostaglandin treatment. CLINICAL RELEVANCE: Immunoreactivity of MMPs in tissues of the monkey uveoscleral outflow pathway is increased after topical treatment with PGF(2 alpha)-IE. This response also might be involved in the intraocular pressure--lowering effect of other prostanoids used to treat glaucoma. PMID- 11483085 TI - Effects of protein kinase inhibitor, HA1077, on intraocular pressure and outflow facility in rabbit eyes. AB - OBJECTIVE: To elucidate the roles of protein kinase in regulating the intraocular pressure (IOP) and outflow facility in rabbit eyes. MATERIALS AND METHODS: A protein kinase inhibitor, 1-(5-isoquinolinesulfonyl)-homopiperazine (HA1077), was used. The IOP and the outflow facility were measured before and after topical, intracameral, or intravitreal administration of HA1077 in rabbits. Western blot analysis was performed to detect the 20-kd light chain of myosin in human trabecular meshwork (TM) cells and bovine ciliary muscle (CM) tissues. The cell morphologic condition and distribution of actin filaments and vinculin in TM cells were studied using cell biology techniques. Carbachol-induced contraction of isolated bovine CM strips following administration of HA1077 was examined in a perfusion chamber. RESULTS: In rabbit eyes, the administration of HA1077 resulted in a significant decrease in IOP in a dose-dependent manner. An increased outflow facility was also observed. Western blot analysis revealed the presence of 20-kd light chain of myosin in human TM cells and bovine CM tissues. In cultured human TM cells, exposure to HA1077 disrupted actin bundles and impaired focal adhesion formation. In addition HA1077 showed relaxation of bovine CM strips. CONCLUSIONS: Use of HA1077 caused a reduction in IOP and an increase in the outflow facility. The results of in vitro experiments suggest that the IOP-lowering effects of HA1077 may be related to the altered cellular behavior of TM cells and relaxation of CM contraction. The results of these studies suggested that protein kinase inhibitors have the potential to be developed into a treatment modality for glaucoma. PMID- 11483086 TI - Real-time optical coherence tomography of the anterior segment at 1310 nm. AB - BACKGROUND: Recent advances in high-speed scanning technology have enabled a new generation of optical coherence tomographic (OCT) systems to perform imaging at video rate. Here, a handheld OCT probe capable of imaging the anterior segment of the eye at high frame rates is demonstrated for the first time. OBJECTIVE: To demonstrate real-time OCT imaging of anterior segment structures. DESIGN: Survey of anterior segment structures in normal human subjects. SETTING: Laboratory. MAIN OUTCOME MEASURES: Achieving real-time imaging of the anterior segment, satisfactory image quality, and convenience of a handheld probe. RESULTS: Optical coherence tomographic imaging of the anterior segment of the eyes of human subjects was performed using 1310-nm wavelength light with an image rate of 8 frames per second. Imaging trials demonstrated clear resolution of corneal epithelium and stroma, sclerocorneal junction, sclera, iris pigment epithelium and stroma, and anterior lens capsule. The anterior chamber angle was clearly visualized. Limited imaging of the ciliary body was performed. Real-time imaging of pupillary constriction in response to light stimulus was also performed. CONCLUSION: High-speed OCT at 1310-nm wavelength is a potentially useful technique for noninvasive assessment of anterior segment structures. CLINICAL RELEVANCE: Our results suggest that real-time OCT has potential applications in glaucoma evaluation and refractive surgery. PMID- 11483087 TI - Visual impairment, age-related cataract, and mortality. AB - OBJECTIVE: To explore associations between visual impairment, cataract, and mortality in older persons after adjusting for other factors associated with mortality. METHODS: A population cohort of 3654 persons aged 49 years or older (82.4% of eligible residents in the Blue Mountains region, west of Sydney, Australia), were examined at the Blue Mountains Eye Study baseline period (1992 1994) and followed up 5 years later (1997-1999). Australian National Death Index data were used to confirm persons who had died since baseline. Associations between mortality and presence of visual impairment and cataract at baseline were assessed using the Cox proportional hazards regression model, controlling for age, sex, demographic and socioeconomic status, medical history, and health risk behaviors. RESULTS: By June 30, 1999, 604 participants (16.5%) had died. The age- and sex-standardized 7-year cumulative mortality rate was 26% among persons with any visual impairment and 16% in persons without visual impairment. After adjusting for factors found significantly associated with mortality, including age, male sex, low self-rated health, low socioeconomic status, systemic medical conditions, and negative health risk behaviors, the presence at baseline of any visual impairment was independently associated with increased mortality risk (risk ratio [RR], 1.7; 95% confidence interval, 1.2-2.3). The presence of age related cataract, either nuclear (RR, 1.5), cortical (RR, 1.3), or posterior subcapsular cataract (RR, 1.5), was also significantly associated with increased mortality risk. These associations remained statistically significant when visual impairment and each type of cataract were included simultaneously in the multivariate Cox model. CONCLUSION: Visual impairment and age-related cataract may be independent risk factors for increased mortality in older persons. PMID- 11483088 TI - Dietary fat and risk for advanced age-related macular degeneration. AB - OBJECTIVE: To evaluate the relationship between intake of total and specific types of fat and risk for advanced age-related macular degeneration (AMD), the leading cause of irreversible blindness in adults. DESIGN: A multicenter eye disease case-control study. SETTING: Five US clinical ophthalmology centers. PATIENTS: Case subjects included 349 individuals (age range, 55-80 years) with the advanced, neovascular stage of AMD diagnosed within 1 year of their enrollment into the study who resided near a participating clinical center. Control subjects included 504 individuals without AMD but with other ocular diseases. Controls were from the same geographic areas as cases and were frequency-matched to cases by age and sex. MAIN OUTCOME MEASURES: Relative risk for AMD according to level of fat intake, controlling for cigarette smoking and other risk factors. RESULTS: Higher vegetable fat consumption was associated with an elevated risk for AMD. After adjusting for age, sex, education, cigarette smoking, and other risk factors, the odds ratio (OR) was 2.22 (95% confidence interval [CI], 1.32-3.74) for persons in the highest vs those in the lowest quintiles of intake (P for trend,.007). The risk for AMD was also significantly elevated for the highest vs lowest quintiles of intake of monounsaturated (OR, 1.71) and polyunsaturated (OR, 1.86) fats (Ps for trend,.03 and.03, respectively). Higher consumption of linoleic acid was also associated with a higher risk for AMD (P for trend,.02). Higher intake of omega-3 fatty acids was associated with a lower risk for AMD among individuals consuming diets low in linoleic acid, an omega-6 fatty acid (P for trend,.05; P for continuous variable,.03). Similarly, higher frequency of fish intake tended to reduce risk for AMD when the diet was low in linoleic acid (P for trend,.05). Conversely, neither omega-3 fatty acids nor fish intake were related to risk for AMD among people with high levels of linoleic acid intake. CONCLUSION: Higher intake of specific types of fat--including vegetable, monounsaturated, and polyunsaturated fats and linoleic acid--rather than total fat intake may be associated with a greater risk for advanced AMD. Diets high in omega-3 fatty acids and fish were inversely associated with risk for AMD when intake of linoleic acid was low. PMID- 11483089 TI - Ten-year follow-up from the CRYO-ROP study. PMID- 11483090 TI - A blind spot in the training of non-English-speaking foreign fellows. PMID- 11483091 TI - Ophthalmic pathology at the crossroads. PMID- 11483092 TI - Lymphoepitheliomalike carcinoma of the orbit. PMID- 11483093 TI - Orbital cavitary rhabdomyosarcoma masquerading as lymphangioma. PMID- 11483094 TI - Uveal effusion and secondary angle-closure glaucoma associated with topiramate use. PMID- 11483095 TI - Ghost cell glaucoma related to snake poisoning. PMID- 11483096 TI - Recurrent transient visual loss after deep sclerectomy. PMID- 11483097 TI - Anterior ischemic optic neuropathy following acute angle-closure glaucoma. PMID- 11483098 TI - Contractile peripapillary staphyloma with light stimulus to the contralateral eye. PMID- 11483099 TI - Shrinkage: fact or fiction? PMID- 11483100 TI - Ciliary body leiomyoma with anterior chamber invasion. PMID- 11483101 TI - Successful closure of spontaneous scleral fistula in retinochoroidal coloboma. PMID- 11483102 TI - In favor of the triple procedure. PMID- 11483105 TI - Removal of emboli from the branches of the central retinal artery. PMID- 11483107 TI - Three questions on the role of sleep apnea syndrome in optic disc edema. PMID- 11483109 TI - Compliance with amblyopia therapy. PMID- 11483110 TI - Why is sun protection in children virtually ignored? PMID- 11483111 TI - When smoke alarms are a nuisance: a call to action. PMID- 11483112 TI - Advocating for children's health at the state level: lessons learned. AB - This article documents the successful creation and promotion of a bill to fund a nurse home visitation program for high-risk mothers in Arkansas. It illustrates several key factors in successful advocacy by pediatricians working in an academic setting: a realistic time commitment; a community needs assessment, data assimilation, and review of existing resources; the identification and incorporation of stakeholders; a narrow focus on the area of greatest need; the backing of political partners; and favorable opportunities to advance child health issues. PMID- 11483114 TI - Pediatric length of stay guidelines and routine practice: the case of Milliman and Robertson. AB - BACKGROUND: Guidelines for inpatient length of stay (LOS) have been developed by Milliman and Robertson (M&R) and are widely applied by health plans. This study was designed to compare LOS for several pediatric conditions with the M&R LOS criteria using recent data and to determine if concordance of actual practice with M&R LOS criteria varied between children and adults. DESIGN: Administrative data from Pennsylvania hospitals from 1996 through 1998 were used to examine LOS for hospital discharges for 12 selected diagnoses for which M&R published guidelines for children and adults. PATIENTS: Discharge data for all patients discharged from public and private hospitals in Pennsylvania for which 1 of 12 selected diagnoses were examined. MAIN OUTCOME MEASURE: Length of stay. RESULTS: In Pennsylvania hospitals from 1996 through 1998, pediatric LOS was divergent for all conditions examined, although not to the extent found in a previous study examining data from New York State. Of note, median LOS for some conditions was shorter than M&R LOS criteria. The percentage of pediatric hospital discharges that exceeded the M&R LOS criteria ranged from 25% for pneumonia to 84% for meningitis. Adult hospital discharges exceeded M&R LOS criteria to a greater extent than did pediatric discharges for all conditions except for sickle cell crisis and meningitis. CONCLUSIONS: The M&R LOS criteria were divergent from routine practice for both children and adults. Greater divergence of adult discharges illustrates the need to consider comorbid conditions when implementing these guidelines. Thus, patient care may suffer if guidelines are implemented in an uninformed way. These findings emphasize the importance of using the best possible science when producing guidelines such as these. PMID- 11483115 TI - Sun protection practices for children: knowledge, attitudes, and parent behaviors. AB - OBJECTIVE: To examine the frequency with which sun protection is used by parents for their children. DESIGN AND SETTING: Descriptive survey conducted at a university medical clinic in Florida. PARTICIPANTS: Parents of children aged 1 to 16 years were approached in the waiting area, and 77 of 100 were successfully interviewed. MAIN OUTCOME MEASURES: Parents' self-reported use of sun protection measures for their children and their attitudes and beliefs about sun protection. RESULTS: Fewer than half of respondents (43%) reported regularly using sun protection for their child. Regular use of sun protection was reported more frequently by female caretakers and those with more favorable attitudes regarding sun protection use. Sunscreen was the most frequently used measure, and preventing sunburn was the primary reason for using sun protection. Respondents held several unfavorable sun protection attitudes, including the belief that sun exposure was healthy, that children looked better with a tan, and that it was okay to stay out in the sun longer if the child wore sunscreen. CONCLUSIONS: Regular use of sun protection for children is infrequent and consists primarily of applying sunscreen rather than methods that reduce sun exposure. Parents primarily use sunscreen to prevent sunburn and may increase their children's overall sun exposure as a result. PMID- 11483116 TI - Psychosocial correlates of physical activity in healthy children. AB - BACKGROUND: Understanding the determinants of physical activity in children is critical for the treatment and prevention of childhood obesity. Social-cognitive theory has been used to understand behavioral patterns in children. OBJECTIVES: To explore the relationship between health beliefs, self-efficacy, social support, and sedentary activities and physical activity levels in children and to examine the relationship between physical activity and children's self-esteem. METHODS: Ninety-two children aged 10 to 16 years completed the study. Physical activity was monitored for 1 week with a motion detector (Actitrac; IM Systems, Baltimore, Md). Moderate-level activity and high-level activity were defined based on the results of treadmill testing. Health beliefs, self-efficacy, social influences, and time spent in sedentary behaviors were determined through questionnaires. Self-esteem was measured using the Piers-Harris Children's Self Concept Scale. Chronic anxiety was measured with the Revised Children's Manifest Anxiety Scale. RESULTS: There was a significant decline in physical activity levels between ages 10 and 16 years, particularly in girls. Preteen girls spent approximately 35% more time in low- and high-level activity than did teenage girls (P<.001). Overall, children spent 75.5% of the day inactive, with a mean +/ SD of 5.2 +/- 1.8 hours watching television, sitting at the computer, and doing homework. In contrast, only 1.4% of the day (12.6 +/- 12.2 minutes) was spent in vigorous activity. Time spent in sedentary behaviors was inversely correlated with the amount of moderate-level activity (P<.001) but not high-level activity. In contrast, time spent in high-level activity correlated with self-efficacy scores (P<.001) and social influences scores (P<.005). High-level physical activity was also associated with improved self-esteem (P<.05). Higher health beliefs scores were not correlated with physical activity levels. CONCLUSIONS: Children and adolescents are largely sedentary. Correlates of high- and low-level physical activity are different. Time spent on sedentary activities is inversely correlated with moderate-level activity, while self-efficacy and social influences are positively correlated with more intense physical activity. In addition, increased high-level physical activity is an important component in the development of self-esteem in children. PMID- 11483117 TI - Serious injuries and deaths of adolescent girls resulting from interpersonal violence: characteristics and trends from the United States, 1989-1998. AB - BACKGROUND: Little published data are available concerning the death and disability of adolescent girls resulting from interpersonal violence (adolescents are defined as those aged 12-18 years in this study). OBJECTIVES: To determine whether there were sex differences in (a) the characteristics of those who were injured or died, (b) injury severity and outcomes, and (c) injury mechanism; and to describe time trends in these differences. DESIGN: Analysis of data concerning serious injuries due to assaults, recorded in the National Pediatric Trauma Registry (from January 1, 1989, through December 31, 1998), and homicides, recorded in the Web-Based Injury Statistics and Query Reporting System database (from January 1, 1990, through December 31, 1997). SETTING: Patient data from participating pediatric trauma centers (National Pediatric Trauma Registry) in 45 states and national death certificate data (Web-Based Injury Statistics and Query Reporting System). PATIENTS: Six hundred twelve adolescent girls who were seriously injured because of an assault were compared with 2656 adolescent boys who were seriously injured because of an assault. Three thousand four hundred eighty-seven adolescent girls who died due to a homicide were compared with 17 292 adolescent boys who died due to a homicide. RESULTS: Assaulted adolescent girls were more likely to have preexisting cognitive or psychosocial impairments than were adolescent boys (odds ratio, 1.68; 95% confidence interval, 1.12-2.51). Adolescent girls trended toward more injury-related impairments at discharge from the hospital (odds ratio, 1.16; 95% confidence interval, 0.92-1.47). Adolescent girls were more likely to have been stabbed, and less likely to have been shot. Also, adolescent girls were more likely to have been injured at a home or a residence. Compared with all National Pediatric Trauma Registry admissions, assaults declined at the same rate for adolescent girls and boys. The proportion resulting from penetrating trauma declined more slowly for adolescent girls. CONCLUSIONS: Interpersonal violence causes considerable morbidity and mortality for young women. Research and interventions should be developed to respond to adolescent girls who experience interpersonal violence. PMID- 11483118 TI - Economic analysis of a child vaccination project among Asian Americans in Philadelphia, Pa. AB - OBJECTIVE: To ascertain the cost-effectiveness and the benefit-cost ratios of a community-based hepatitis B vaccination catch-up project for Asian American children conducted in Philadelphia, Pa, from October 1, 1994, to February 11, 1996. DESIGN: Program evaluation. SETTING: South and southwest districts of Philadelphia. PARTICIPANTS: A total of 4384 Asian American children. INTERVENTIONS: Staff in the community-based organizations (1) educated parents about the hepatitis B vaccination, (2) enrolled physicians in the Vaccines for Children program, and (3) visited homes of children due for a vaccine dose. Staff in the Philadelphia Department of Public Health developed a computerized database; sent reminder letters for children due for a vaccine dose; and offered vaccinations in public clinics, health fairs, and homes. MAIN OUTCOME MEASURES: The numbers of children having received 1, 2, or 3 doses of vaccine before and after the interventions; costs incurred by the Philadelphia Department of Public Health and the community-based organizations for design, education, and outreach activities; the cost of the vaccination; cost-effectiveness ratios for intermediate outcomes (ie, per child, per dose, per immunoequivalent patient, and per completed series); discounted cost per discounted year of life saved; and the benefit-cost ratio of the project. RESULTS: For the completed series of 3 doses, coverage increased by 12 percentage points at a total cost of $268 660 for design, education, outreach, and vaccination. Costs per child, per dose, and per completed series were $64, $119, and $537, respectively. The discounted cost per discounted year of life saved was $11 525, and 106 years of life were saved through this intervention. The benefit-cost ratio was 4.44:1. CONCLUSION: Although the increase in coverage was modest, the intervention proved cost effective and cost-beneficial. PMID- 11483119 TI - Hepatitis B vaccination practices in hospital newborn nurseries before and after changes in vaccination recommendations. AB - BACKGROUND: Routine use of hepatitis B vaccine for low-risk newborns was suspended on July 7, 1999, because of concern about the potential risk of thimerosal, a mercury-containing vaccine preservative. Reinstatement of the birth dose was recommended when a thimerosal-free vaccine became available. OBJECTIVE: To explore changes in hepatitis B vaccination practices for newborns related to the revised recommendations for low-risk infants (in this study, the terms newborn and infant are used interchangeably). DESIGN: A telephone survey of a random sample of 1000 US hospitals. PARTICIPANTS: Nurse managers, nursery directors, and staff nurses of the newborn nurseries. MAIN OUTCOME MEASURES: Nursery vaccination practices before and after July 7, 1999, and the availability and use of thimerosal-free vaccine. RESULTS: Interviews were conducted with 773 (87%) of 886 eligible hospitals. Before July 7, 1999, 78% of the hospitals reported vaccination practices that were consistent with recommendations at that time, although only 47% vaccinated all low-risk infants at birth. After July 7, 1999, almost all hospitals discontinued vaccination of low-risk infants, in accordance with the recommendation change; however, there was a 6-fold increase in the number of hospitals that were not vaccinating all high-risk infants. After the introduction of thimerosal-free vaccine, only 39% of the hospitals reported vaccinating all low-risk infants. CONCLUSIONS: Most hospital nurseries altered their newborn hepatitis B vaccination practices consistent with changes in national recommendations. However, unintended consequences included the failure of some hospitals to continue vaccinating all high-risk infants and the delay in reintroducing vaccination for low-risk newborns after the introduction of a thimerosal-free vaccine. Assessments of the appropriateness of this country's response to the threat of thimerosal in vaccines should consider these findings. PMID- 11483120 TI - Correlates of vaccination for hepatitis B among adolescents: results from a parent survey. AB - OBJECTIVE: To identify correlates of hepatitis B vaccination status in sixth grade students in the year prior to implementation of a requirement mandating immunization for seventh-grade entry. METHODS: A survey of parents of sixth graders in 5 schools in San Diego County. Two logistic regression models were tested to predict the outcome variables: initiation and completion of the vaccination series. RESULTS: Factors associated with initiating the series included a recent nonacute medical visit, white race, hearing about the vaccination law from a health care provider, and the availability of a school based vaccination clinic. Factors associated with completing the series included English as the primary language spoken at home, hearing about the law from a health care provider, a school-based vaccination clinic, and higher socioeconomic status. Health insurance was not significantly related to either outcome. CONCLUSIONS: There was a clear benefit for hepatitis B immunization status to have had a recent nonacute medical visit, to have heard about the law from a health care provider, and to have a school-based vaccination clinic. The factors associated with starting vs completing the vaccination series were not identical. However, both health care provider and school characteristics were related to starting and completing the vaccination series. Thus, a multifaceted strategy may be most appropriate for successful coverage of an adolescent population with a vaccination series such as hepatitis B. PMID- 11483121 TI - When parents reject interventions to reduce postnatal human immunodeficiency virus transmission. AB - In a recent Oregon case, the state successfully sued for custody of an infant to prevent his human immunodeficiency virus (HIV)-infected mother from breastfeeding him and to require antiretroviral prophylaxis. As more HIV-infected women give birth, pediatricians may increasingly face dilemmas when parents reject medical recommendations to forego breastfeeding and to administer antiretroviral prophylaxis to the infant. Such disagreements create ethical dilemmas because pediatricians have an obligation to both protect the infant and respect parental decision making. Pediatricians need to balance these obligations in deciding whether to ask the courts to intervene on the infant's behalf. To that end, we analyze the legal and ethical issues that arise when an HIV-infected mother refuses interventions to reduce neonatal transmission of HIV to her infant, provide an approach for addressing these disagreements, and present illustrative scenarios in which pediatricians should, may, and should not seek a court order to intervene. PMID- 11483122 TI - Theophylline toxicokinetics in premature newborns. AB - BACKGROUND: While cytochrome P4501A2 is the primary pathway for theophylline (aminophylline ethylenediamine) metabolism in adults, it is developmentally immature in the newborn. OBJECTIVE: To report the developmental differences in theophylline toxicokinetics of neonates. DESIGN: Case series. Three premature neonates received inadvertent intravenous overdoses of theophylline for apnea of prematurity while in newborn intensive care. Maximum serum concentrations ranged from 55 to 123 mg/L. Theophylline-derived caffeine levels plateaued at 8.4 to 13 mg/L and did not decline during the sampling period. All newborns experienced sinus tachycardia and agitation. Sequential theophylline and caffeine serum levels were obtained periodically for 62 to 100 hours. In contrast to older children and adults, in whom theophylline disposition follows zero-order kinetics at high concentrations, a monoexponential function best described theophylline elimination in the premature newborn, with half-lives ranging from 24.7 to 36.5 hours and estimated clearance from 0.02 to 0.05 L/kg per hour. These values are consistent with those previously reported in neonates. All patients were treated with supportive care without invasive procedures. No seizures or apparent sequelae occurred. CONCLUSION: Developmental differences in the balance between nonrenal (ie, metabolic) and renal elimination pathways produce the unique toxicokinetics of theophylline in the neonate. PMID- 11483123 TI - A multicomponent program for nutrition and physical activity change in primary care: PACE+ for adolescents. AB - BACKGROUND: Most adolescents do not meet national recommendations for nutrition and physical activity. However, no studies of physical activity and nutrition interventions for adolescents conducted in health care settings have been published. The present study was an initial evaluation of the PACE+ (Patient centered Assessment and Counseling for Exercise plus Nutrition) program, delivered in primary care settings. PARTICIPANTS: Adolescents aged 11 to 18 years (N = 117) were recruited from 4 pediatric and adolescent medicine outpatient clinics. Participants' mean (SD) age was 14.1 (2.0) years, 37% were girls, and 43% were ethnic minorities. INTERVENTION: Behavioral targets were moderate physical activity, vigorous physical activity, fat intake, and fruit and vegetable intake. All patients completed a computerized assessment, created tailored action plans to change behavior, and discussed the plans with their health care provider. Patients were then randomly assigned to receive no further contact or 1 of 3 extended interventions: mail only, infrequent telephone and mail, or frequent telephone and mail. MEASURES: Brief, validated, self-report measures of target behaviors were collected at baseline and 4 months later. RESULTS: All outcomes except vigorous physical activity improved over time, but adolescents who received the extended interventions did not have better 4-month outcomes than those who received only the computer and provider counseling components. Adolescents who targeted a behavior tended to improve more than those who did not target the behavior, except for those who targeted vigorous physical activity. CONCLUSIONS: A primary care-based interactive health communication intervention to improve physical activity and dietary behaviors among adolescents is feasible. Controlled experimental research is needed to determine whether this intervention is efficacious in changing behaviors in the short- and long-term. PMID- 11483124 TI - Subsequent sexually transmitted infection in urban adolescents and young adults. AB - OBJECTIVE: To compare the rates of subsequent infection with Chlamydia trachomatis, Neisseria gonorrhoeae, or Trichomonas vaginalis in a group of high risk adolescents and young adults. METHODS: At the time of treatment, 444 unmarried teenagers and young adults aged 13 to 25 years were enrolled from an urban sexually transmitted disease clinic and 3 community-based primary care clinics. Subjects were infected with C trachomatis, N gonorrhoeae, or T vaginalis, were diagnosed as having nongonococcal urethritis (in men), or were uninfected sexual contacts with one of these infections. Subjects returned at 1, 3, 5, and 7 months. RESULTS: The rate of subsequent infection was substantial. Forty percent of men and 53% of women who were uninfected contacts at enrollment were estimated to be infected within 7 months; 60% of men and 73% of women infected at enrollment were estimated to be reinfected. Among women, subjects who were infected at enrollment had a shorter time to subsequent infection (median, 140 days) compared with uninfected contacts (median, 209 days) (P =.04). Among men, findings were similar, but the difference in median time to subsequent infection was not significant (P =.08). Baseline characteristics that predicted shorter time to reinfection were female sex and infection at enrollment. When sexual behaviors in the 2 months preceding each subsequent data collection visit were included in the model, only being female and reporting at least one new interval sexual partner were significant predictors of subsequent sexually transmitted infections. CONCLUSIONS: These data support recent research that has found high rates of subsequent infection among high-risk adolescents and young adults. Contacts of a sexually transmitted infection appear to be at equally high risk for subsequent infection as those with a personal history of infection. Our data suggest that more frequent than annual screening for N gonorrhoeae, C trachomatis, and T vaginalis would be appropriate in at-risk adolescent and young adult populations, including individuals who are uninfected sexual contacts to a sexually transmitted infection. PMID- 11483125 TI - Telephone subsidy: an effective incentive for successful participation in home memory monitor study. AB - CONTEXT: The Collaborative Home Infant Monitoring Evaluation (CHIME) study enrolled healthy term infants and 3 groups of infants considered to be at increased risk for sudden infant death syndrome to evaluate apnea and bradycardia events in the home. Mother-infant pairs without a telephone were ineligible for enrollment. OBJECTIVE: To determine whether mother-infant pairs who were offered a telephone subsidy would agree to enroll in CHIME and achieve protocol compliance rates comparable with those of matched subjects able to afford telephones. DESIGN: Thirty-one telephone subsidy subjects were retrospectively compared with 55 control subjects matched for study group, site, birth weight, and maternal race, age, and education. SETTING: Collaborative Home Infant Monitoring Evaluation clinical research centers in Honolulu, Hawaii, and Toledo, Ohio. INTERVENTION: Provision of telephone subsidy to otherwise eligible enrollees for CHIME. MAIN OUTCOME MEASURES: Frequency of compliance with protocol requirements for follow-up evaluations and for extent of home monitoring. RESULTS: Subsidy subjects achieved protocol completion rates that were comparable with those of control subjects, for developmental assessments at 56 and 92 weeks postconceptional age (PCA), and for the polysomnogram. Unexpectedly, however, subsidy subjects were more likely to have a developmental assessment at 44 weeks PCA (P =.02), as well as a cry analysis (P =.04). They were also more likely to use the CHIME home monitor for more hours during weeks 2 through 5 (P =.004), have a higher percentage using the monitor for 10 or more hours per week during weeks 2 through 5 (P =.009), and have a higher total number of days of monitor use throughout 6 months (P <.001). Mean cost of the subsidy was $3.25 per day of monitor use, and monitor use per day was directly related to total cost of the subsidy (P =.02). CONCLUSIONS: Telephone subsidy is an effective financial incentive. At least within the context of the CHIME study, telephone subsidy enhanced access to health care, and in some categories it resulted in enhanced protocol compliance. PMID- 11483126 TI - Picture of the month: cardiac rhabdomyoma in tuberous sclerosis. PMID- 11483127 TI - Radiological case of the month: pulmonary air embolus with home antibiotic infusion. PMID- 11483128 TI - Radiological case of the month: Lemierre syndrome. PMID- 11483129 TI - Pathological case of the month: classic rickets in a setting of significant psychosocial deprivation. PMID- 11483130 TI - Pathological case of the month: Desbuquois syndrome. PMID- 11483131 TI - Race and ethnicity in biomedical and health services research. PMID- 11483132 TI - Constructive use of race and ethnicity variables. PMID- 11483133 TI - Race not always useful in final analysis. PMID- 11483135 TI - Resident autonomy in an era of managed care. PMID- 11483137 TI - Psychiatric disorders and drug use among human immunodeficiency virus-infected adults in the United States. AB - BACKGROUND: There have been no previous nationally representative estimates of the prevalence of mental disorders and drug use among adults receiving care for human immunodeficiency virus (HIV) disease in the United States. It is also not known which clinical and sociodemographic factors are associated with these disorders. SUBJECTS AND METHODS: We enrolled a nationally representative probability sample of 2864 adults receiving care for HIV in the United States in 1996. Participants were administered a brief structured psychiatric instrument that screened for psychiatric disorders (major depression, dysthymia, generalized anxiety disorders, and panic attacks) and drug use during the previous 12 months. Sociodemographic and clinical factors associated with screening positive for any psychiatric disorder and drug dependence were examined in multivariate logistic regression analyses. RESULTS: Nearly half of the sample screened positive for a psychiatric disorder, nearly 40% reported using an illicit drug other than marijuana, and more than 12% screened positive for drug dependence during the previous 12 months. Factors independently associated with screening positive for a psychiatric disorder included number of HIV-related symptoms, illicit drug use, drug dependence, heavy alcohol use, and being unemployed or disabled. Factors independently associated with screening positive for drug dependence included having many HIV-related symptoms, being younger, being heterosexual, having frequent heavy alcohol use, and screening positive for a psychiatric disorder. CONCLUSIONS: Many people infected with HIV may also have psychiatric and/or drug dependence disorders. Clinicians may need to actively identify those at risk and work with policymakers to ensure the availability of appropriate care for these treatable disorders. PMID- 11483138 TI - Use of mental health and substance abuse treatment services among adults with HIV in the United States. AB - BACKGROUND: The need for mental health and substance abuse services is great among those with human immunodeficiency virus (HIV), but little information is available on services used by this population or on individual factors associated with access to care. METHODS: Data are from the HIV Cost and Services Utilization Study, a national probability survey of 2864 HIV-infected adults receiving medical care in the United States in 1996. We estimated 6-month use of services for mental health and substance abuse problems and examined socioeconomic, HIV illness, and regional factors associated with use. RESULTS: We estimated that 61.4% of 231 400 adults under care for HIV used mental health or substance abuse services: 1.8% had hospitalizations, 3.4% received residential substance abuse treatment, 26.0% made individual mental health specialty visits, 15.2% had group mental health treatment, 40.3% discussed emotional problems with medical providers, 29.6% took psychotherapeutic medications, 5.6% received outpatient substance abuse treatment, and 12.4% participated in substance abuse self-help groups. Socioeconomic factors commonly associated with poorer access to health services predicted lower likelihood of using mental health outpatient care, but greater likelihood of receiving substance abuse treatment services. Those with less severe HIV illness were less likely to access services. Persons living in the Northeast were more likely to receive services. CONCLUSIONS: The magnitude of mental health and substance abuse care provided to those with known HIV infection is substantial, and challenges to providers should be recognized. Inequalities in access to care are evident, but differ among general medical, specialty mental health, and substance abuse treatment sectors. PMID- 11483139 TI - Emotion in criminal offenders with psychopathy and borderline personality disorder. AB - BACKGROUND: Criminal offenders with a diagnosis of psychopathy or borderline personality disorder (BPD) share an impulsive nature but tend to differ in their style of emotional response. This study aims to use multiple psychophysiologic measures to compare emotional responses to unpleasant and pleasant stimuli. METHODS: Twenty-five psychopaths as defined by the Hare Psychopathy Checklist and 18 subjects with BPD from 2 high-security forensic treatment facilities were included in the study along with 24 control subjects. Electrodermal response was used as an indicator of emotional arousal, modulation of the startle reflex as a measure of valence, and electromyographic activity of the corrugator muscle as an index of emotional expression. RESULTS: Compared with controls, psychopaths were characterized by decreased electrodermal responsiveness, less facial expression, and the absence of affective startle modulation. A higher percentage of psychopaths showed no startle reflex. Subjects with BPD showed a response pattern very similar to that of controls, ie, they showed comparable autonomic arousal, and their startle responses were strongest to unpleasant slides and weakest to pleasant slides. However, corrugator electromyographic activity in subjects with BPD demonstrated little facial modulation when they viewed either pleasant or unpleasant slides. CONCLUSIONS: The results support the theory that psychopaths are characterized by a pronounced lack of fear in response to aversive events. Furthermore, the results suggest a general deficit in processing affective information, regardless of whether stimuli are negative or positive. Emotional hyporesponsiveness was specific to psychopaths, since results for offenders with BPD indicate a widely adequate processing of emotional stimuli. PMID- 11483140 TI - Obstetric complications, parenting, and risk of criminal behavior. AB - BACKGROUND: The results of studies that have examined the relationship between prenatal and perinatal complications and adult criminality and violence are contradictory. Supporting evidence for this relationship comes from studies of samples drawn from a single cohort. The present study was designed to examine the associations between prenatal and perinatal complications and criminality, defining more precisely than past investigations subject characteristics and the types of offenses. METHODS: The cohort includes the 15 117 persons born in Stockholm, Sweden, in 1953 and followed up to age 30 years. Information was extracted from obstetric files, health, social, work, and criminal records. Obstetric complications were defined as deviations from normal development occurring at any point from conception through the neonatal period. Inadequate parenting was indexed by social intervention. RESULTS: Inadequate parenting was experienced by 19.1% of the men and 18.1% of the women, and was shown to increase the risk of offending (men, 1.39 times [95% confidence interval [CI], 1.28-1.50]; women, 2.09 [95% CI, 1.70-2.56]) and of violent offending (men, 2.02 times [95% CI, 1.67-2.44]; women, 2.09 [95% CI, 1.70-2.56]). Obstetric complications in the absence of family problems did not increase the risk of offending. A combination of pregnancy complications and inadequate parenting affected 3.1% of the men and 4.0% of the women, and increased the risk of offending (1.64 times [95% CI, 1.43 1.89]; 1.79 times [95% CI, 1.16-2.75], respectively) and violent offending (2.86 times [95% CI, 2.09-3.91]; 1.81 times [95% CI, 0.57-5.79]). CONCLUSIONS: A combination of pregnancy complications and inadequate parenting increased the risk of violent and nonviolent offending only slightly more than inadequate parenting alone. However, inadequate parenting was experienced by 5 times more cohort members than was the combination of inadequate parenting and pregnancy complications. PMID- 11483141 TI - Targeting behavioral therapies to enhance naltrexone treatment of opioid dependence: efficacy of contingency management and significant other involvement. AB - BACKGROUND: Contingency management (CM) and significant other involvement (SO) were evaluated as strategies to enhance treatment retention, medication compliance, and outcome for naltrexone treatment of opioid dependence. METHODS: One hundred twenty-seven recently detoxified opioid-dependent individuals were randomly assigned to 1 of 3 conditions delivered for 12 weeks: (1) standard naltrexone treatment, given 3 times a week; (2) naltrexone treatment plus contingency management (CM), with delivery of vouchers contingent on naltrexone compliance and drug-free urine specimens; or (3) naltrexone treatment, CM, plus significant other involvement (SO), where a family member was invited to participate in up to 6 family counseling sessions. Principal outcomes were retention in treatment, compliance with naltrexone therapy, and number of drug free urine specimens. RESULTS: First, CM was associated with significant improvements in treatment retention (7.4 vs 5.6 weeks; P =.05) and in reduction in opioid use (19 vs 14 opioid-free urine specimens; P =.04) compared with standard naltrexone treatment. Second, assignment to SO did not significantly improve retention, compliance, or substance abuse outcomes compared with CM. Significant effects for the SO condition over CM on retention, compliance, and drug use outcomes were seen only for the subgroup who attended at least 1 family counseling session. The SO condition was associated with significant (P =.02) improvements in family functioning. CONCLUSION: Behavioral therapies, such as CM, can be targeted to address weaknesses of specific pharmacotherapies, such as noncompliance, and thus can play a substantial role in broadening the utility of available pharmacotherapies. PMID- 11483142 TI - Neurocognitive enhancement therapy with work therapy: effects on neuropsychological test performance. AB - BACKGROUND: Cognitive deficits are a major determinant of social and occupational dysfunction in schizophrenia. In this study, we determined whether neurocognitive enhancement therapy (NET) in combination with work therapy (WT) would improve performance on neuropsychological tests related to but different from the training tasks. METHODS: Sixty-five patients with schizophrenia or schizoaffective disorder were randomly assigned to NET plus WT or WT alone. Neurocognitive enhancement therapy included computer-based training on attention, memory, and executive function tasks; an information processing group; and feedback on cognitive performance in the workplace. Work therapy included paid work activity in job placements at the medical center (eg, mail room, grounds, library) with accompanying supports. Neuropsychological testing was performed at intake and 5 months later. RESULTS: Prior to enrollment, both groups did poorly on neuropsychological testing. Patients receiving NET + WT showed greater improvements on pretest-posttest variables of executive function, working memory, and affect recognition. As many as 60% in the NET + WT group improved on some measures and were 4 to 5 times more likely to show large effect-size improvements. The number of patients with normal working memory performance increased significantly with NET + WT, from 45% to 77%, compared with a decrease from 56% to 45% for those receiving WT. CONCLUSIONS: Computer training for cognitive dysfunction in patients with schizophrenia can have benefits that generalize to independent outcome measures. Efficacy may result from a synergy between NET, which encourages mental activity, and WT, which allows a natural context for mental activity to be exercised, generalized, and reinforced. PMID- 11483143 TI - Neural correlates of formal thought disorder in schizophrenia: preliminary findings from a functional magnetic resonance imaging study. AB - BACKGROUND: Formal thought disorder (FTD) is a core symptom of schizophrenia, but its pathophysiology is little understood. We examined the neural correlates of FTD using functional magnetic resonance imaging. METHODS: Blood oxygenation level dependent contrast was measured using functional magnetic resonance imaging while 6 patients with schizophrenia and 6 control subjects spoke about 7 Rorschach inkblots for 3 minutes each. In patients, varying degrees of thought-disordered speech were elicited during each "run." In a within-subject design, the severity of positive FTD was correlated with the level of blood oxygenation level dependent contrast in the 2 runs that showed the highest variance of FTD in each patient. RESULTS: The severity of positive FTD in patients was negatively correlated (P<.001) with signal changes in the left superior and middle temporal gyri. Positive correlations were evident in the cerebellar vermis, the right caudate body, and the precentral gyrus. CONCLUSIONS: The severity of positive FTD was inversely correlated with the level of activity in the Wernicke area, a region implicated in the production of coherent speech. Reduced activity in this area might contribute to the articulation of incoherent speech. Because of the small sample size, these findings should be considered preliminary. PMID- 11483144 TI - Efficacy of a mixed amphetamine salts compound in adults with attention deficit/hyperactivity disorder. AB - BACKGROUND: We report on a controlled trial of a mixed amphetamine salts compound (Adderall, dextroamphetamine sulfate, dextro-, levoamphetamine sulfate, dextroamphetamine aspartate, levoamphetamine aspartate, and dextroamphetamine saccharate) in the treatment of adult attention-deficit/hyperactivity disorder (ADHD). METHODS: This was a 7-week, randomized, double-blind, placebo-controlled, crossover study of Adderall in 27 well-characterized adults satisfying full DSM IV criteria for ADHD of childhood onset and persistent symptoms into adulthood. Medication was titrated up to 30 mg twice a day. Outcome measures included the ADHD Rating Scale and the Clinical Global Impression Score. Comorbid psychiatric disorders were assessed to test for potential effects on treatment outcome. RESULTS: Treatment with Adderall at an average oral dose of 54 mg (administered in 2 daily doses) was effective and well tolerated. Drug-specific improvement in ADHD symptoms was highly significant overall (42% decrease on the ADHD Rating Scale, P<.001), and sufficiently robust to be detectable in a parallel groups comparison restricted to the first 3 weeks of the protocol (P<.001). The percentage of subjects who improved (reduction in the ADHD rating scale of > or =30%) was significantly higher with Adderall treatment than with a placebo (70% vs 7%; P =.001). CONCLUSIONS: Adderall was effective and well tolerated in the short-term treatment of adults with ADHD. More work is needed to evaluate the long-term effects of Adderall, or other amphetamine compounds, in the treatment of adults with ADHD. PMID- 11483145 TI - Attention-deficit/hyperactivity disorder in adults: beyond controversy. PMID- 11483146 TI - A self-report scale to help make psychiatric diagnoses: the Psychiatric Diagnostic Screening Questionnaire. AB - BACKGROUND: The Psychiatric Diagnostic Screening Questionnaire (PDSQ) is a brief, psychometrically strong, self-report scale designed to screen for the most common DSM-IV Axis I disorders encountered in outpatient mental health settings. In the present report, we describe the diagnostic performance (sensitivity, specificity, and positive and negative predictive values) of the PDSQ in an outpatient setting. METHODS: Six hundred thirty psychiatric outpatients presenting for treatment were evaluated with the Structured Clinical Interview for DSM-IV after completing the PDSQ. Patients arrived approximately 20 minutes before the scheduled time of the appointment to complete the scale. Diagnostic raters were blind to responses on the scale. RESULTS: The PDSQ's subscales' diagnostic performance varied in a predictable manner according to the cutoff score-as the threshold for case identification increased, subscale sensitivity decreased and specificity increased. Mean subscale sensitivities of 80%, 85%, and 90% resulted in mean subscale specificities of 78%, 73%, and 66%, respectively, and negative predictive values of 95%, 96%, and 97%. Receiver operating curves were determined for each subscale and all areas under the curve were significant. CONCLUSIONS: The PDSQ is a diagnostic aid designed to be used in clinical practice to facilitate the efficiency of conducting initial diagnostic evaluations. From a clinical perspective, it is most important that a diagnostic aid have good sensitivity, so that most cases are detected, and high negative predictive value, so that most noncases on the measure are indeed noncases. Our results indicate that most of the PDSQ subscales were able to achieve this goal. PMID- 11483147 TI - Cloning and expression of the external-glycoprotein gene mutant from HIV-2 in the methylotrophic yeast Pichia pastoris and identification of the glycoprotein. AB - To achieve high-level expression of HIV-2(ROD) external glycoprotein gp105 in Pichia pastoris, the gp105 gene mutant tP1, with the 5' non-functional region of the gp105 gene removed, was obtained by PCR amplification and was cloned into secreted expression vector pHILS1. The His(+)Mut(s) recombinant P. pastoris strain was screened by PCR and induced by methanol. SDS/PAGE and Western-blot analyses showed that mutation of the low-usage codon AGG into synonymous codon CGA and the introduction of the optimal codon TTC made P. pastoris overexpress tP1, an 85 kDa heterologous glycoprotein that was secreted into the medium and recognized specifically by HIV-2 polyclonal antibody. The recombinant strain GS115/tP1 had excellent genetic stability in terms of the properties of growth and expression of gp105, and seven out of 58 recombinant stains with a yield of 29% were selected to be used for further purification of gp105. PMID- 11483148 TI - Adhesion of tear proteins to contact lenses and vials. AB - Tear-protein adhesion to contact lenses contributes to lens contamination and deterioration, and depends primarily on the type of contact-lens material used. Adhesion of tear proteins to contact lenses and glass vials was examined using spectrophotometry, hydrophobic-interaction HPLC and bicinchoninic acid (BCA) protein assay. Using spectrophotometry at 280 nm, vials containing two proteins (lysozyme and milk lipocalin) at three different concentrations were examined for changes in their protein concentrations over a 3-week period. Vials containing 5.0 mg/ml lysozyme lost about one-third of their protein (P<0.005), while vials containing 5.0 mg/ml milk lipocalin lost two-thirds of their protein (P<0.01). Conversely, vials containing 1.0 mg/ml lysozyme lost two-thirds of their protein (P<0.005), while vials containing 1.0 mg/ml milk lipocalin lost one-third of their protein (P>0.05). Subsequently, lysozyme deposition on both glass vials and contact lenses was monitored for 5 days using spectrophotometry, to determine lysozyme content in the vials, and HPLC, to determine lysozyme deposition on contact lenses stored in the same vials. This experiment indicated considerable variation in lysozyme deposition on vials and contact lenses, with vial deposition remaining relatively constant (P>0.05) while lens deposition decreased and then increased (P<0.05). Finally, the same experiment was repeated, monitoring lysozyme deposition using the BCA assay. This experiment yielded the most consistent results, with lysozyme deposition on vials continuing throughout the 5 day experiment (P<0.05), while deposition on lenses again decreased and then increased, although to a much lesser extent than in the previous experiment (P>0.05). PMID- 11483149 TI - Preparation of stable, highly active and immobilized glucose oxidase using the anti-enzyme antibodies and F(ab)'2. AB - Immobilization of the Aspergillus niger glucose oxidase in high yield was achieved using an immunoaffinity-based procedure. For this purpose IgGs, isolated from the sera of rabbits immunized with glucose oxidase, were favourably oriented by binding on to cobalt-charged iminodiacetate-Sepharose. Large amounts of glucose oxidase could be immobilized by incubating the IgG-bound matrix alternately with the enzyme and either intact IgG or F(ab)2 derived thereof, leading to the formation of multiple enzyme layers. After three incubation cycles using anti-(glucose oxidase) IgG, an 8-fold increase in the amount of enzyme immobilized was observed, while the increase was 11-fold when the F(ab)2 replaced intact IgG. The preparations obtained thus were highly active, as also indicated by the high effectiveness factor, eta. Immunoaffinity-layered immobilized preparations were markedly more resistant to inactivation induced by exposure to 60 degrees C, 4.0 M urea or storage at 4 degrees C. The preparations also exhibited a remarkable resistance against inactivation induced by the water miscible organic solvents tetrahydrofuran, dioxan or acetone. Immobilized glucose oxidase preparations obtained using F(ab)2 were generally observed to be superior in stability compared with those immobilized with the help of intact IgG. PMID- 11483150 TI - Impaired L-arginine uptake in platelets from type-2 diabetic patients. AB - The L-arginine uptake of resting platelets from type-2 diabetic patients and control subjects was measured and the kinetic parameters defined. The effect of platelet stimulation with agonists was also investigated. Kinetic studies showed that the K(m) value for L-arginine transport was not different in patients compared with control subjects, while V(max) was significantly decreased in patients compared with controls. Moreover, agonists able to mobilize Ca(2+) produced a further decrease in the L-arginine uptake in controls and in patients, suggesting that Ca(2+) intracellular levels down-regulate L-arginine transport in platelets from both control subjects and patients. Data suggest that drugs designed to control intracellular Ca(2+) might restore platelet function in diabetic patients. PMID- 11483151 TI - Mammalian cell production of a respiratory syncytial virus (RSV) candidate vaccine recovered using a product-specific affinity column. AB - The recombinant production of a respiratory syncytial virus (RSV) candidate vaccine BBG2Na in baby hamster kidney cells (BHK-21 cells) was investigated. BBG2Na consists of a serum-albumin-binding region (BB) fused to a 101-amino-acid fragment of the RSV G-protein. Semliki Forest virus-based expression vectors encoding both intracellular and secreted forms of BBG2Na were constructed and found to be functional. Affinity recovery of BBG2Na employing human serum albumin columns was found to be inefficient due to the abundance of BSA in the applied samples. Instead, a strategy using a tailor-made affinity ligand based on a combinatorially engineered Staphylococcus aureus protein A domain, showing specific binding to the G-protein part of the product, was evaluated. In conclusion, a strategy for production and successful recovery of BBG2Na in mammalian cells was created, through the development of a product-specific affinity column. PMID- 11483152 TI - A novel method of isolation of a bone-morphogenetic-protein-like protein from ossein. AB - A new bone-morphogenetic-protein (BMP)-like protein has been isolated through a new protocol from a novel source, ossein. The BMP-like protein was hydrophilic and characterized through Fourier-transform IR studies, SDS/PAGE and coupled with a neutral binder, hydroxypropylmethylcellulose (HPMC) for control release. The IR spectrum of the protein showed peaks in tandem with BMP from bone matrix, and its molecular mass was in the range 18-21 kDa. Sustained release from the surface of HPMC was achieved for a period of 3 days. PMID- 11483153 TI - Cloning, sequence analysis and functional characterization of DNA polymerase I from the thermophilic eubacterium Rhodothermus marinus. AB - A gene encoding a DNA polymerase I from the thermophilic eubacterium Rhodothermus marinus was identified. The gene was cloned, sequenced and expressed in Escherichia coli. The gene is 2772 bp long and encodes a protein of 924 amino acids with a calculated molecular mass of 104.8 kDa. Sequence analysis showed that a generally conserved Phe residue in the O-helix is substituted by a Tyr (position 756) in the R. marinus enzyme. A Tyr in this position decreases the discrimination against dideoxynucleotides which is a major advantage in DNA sequencing. The protein was purified, characterized and showed to contain specific DNA-polymerization activity of 3100 units/mg of protein, 5'-->3' exonuclease activity and a 3'-->5' proofreading activity. Its optimum activity was at 55 degrees C and it had a half-life of 2 min at 90 degrees C. A truncated form of the enzyme lacking the 5'-->3' exonuclease domain was also expressed in E. coli. It had a specific DNA-polymerization activity of 5000 units/mg of protein and lacked the 5'-->3' exonuclease activity. Its optimum activity was at 65 degrees C and it had a half-life of 11 min at 90 degrees C. It was usable for DNA sequencing. This is the first thermostable DNA polymerase described with the O-helix Phe-->Tyr substitution. PMID- 11483154 TI - Two telomerase reverse transcriptases (TERTs) expressed in Candida albicans. AB - The human pathogenic yeast Candida albicans contains two telomerase reverse transcriptase (TERT) genes. CaTERT1 and CaTERT2 appear either to be two alleles of the same gene or two entirely different genes that encode 867-residue proteins that differ by five amino acids. Both TERTs have a calculated pI of 9.5 and a M(r) of 100.9 kDa and are the smallest TERTs identified to date. Both genes appear to be expressed. Based on sequence similarity between CaTERT1 and the Saccharomyces cerevisiae orthologue Est2p, we suggest a revised alignment for motif E of Est2p. The identification of these TERT genes provides the first opportunity to study telomerase in an important human pathogen. PMID- 11483155 TI - Characterization of gelatin-immobilized pigeonpea urease and preparation of a new urea biosensor. AB - Urease purified from pigeonpea seeds was immobilized on gelatin beads via cross linking with glutaraldehyde. The maximum immobilization (75%) was observed at 30 mg/ml gelatin, 0.414 mg of enzyme/bead, 1% (v/v) glutaraldehyde and 4 degrees C. Beads stored in 50 mM Tris/acetate buffer (pH 7.3) at 4 degrees C showed a half life of 240 days and there was practically no leaching of enzyme (less than 2%) over a period of 30 days. These beads can be reused more than 30 times (with 24 h intervals) without much loss of enzyme activity (i.e. less than 11%). The immobilized urease showed a shift in its optimum pH from 7.3 to 6.5 in Tris/acetate buffer. Optimum temperature also shifted from 47 to 65 degrees C compared with the soluble enzyme. Gelatin-immobilized pigeonpea urease had a higher K(m) (8.3 mM) than that of the soluble enzyme (3.0 mM). The time-dependent temperature inactivation pattern was also found to change from biphasic to monophasic kinetics. The immobilized beads were used for the preparation of a new urea biosensor with a response time of less than 2 min. At least 14 samples of urea can be measured with this biosensor within an hour. The beads, as well as the biosensor, were used to analyse the urea content in clinical samples from the local clinical pathology laboratories. The results obtained with the biosensor were strikingly similar to those obtained with the various commonly employed biochemical/autoanalyzer(R) methods used. These immobilization studies also have a potential role in haemodialysis machines that maintain the urea level in kidney patients and in the construction of a portable/wearable kidney. The easy availability of the pigeonpea urease, the ease of its immobilization on gelatin and a significantly lower cost of the urease described in the present study makes it a suitable product for future applications in therapeutics and diagnostics. PMID- 11483156 TI - Recombinant Opc protein from Neisseria meningitidis reconstituted into liposomes elicits opsonic antibodies following immunization. AB - The reconstitution of recombinant bacterial outer membrane proteins (OMPs) into their native conformations after purification has been the major problem in their use as effective vaccines. Liposomes have been shown to be an attractive approach, providing a native-like environment for these antigens. The meningococcal recombinant Opc (rOpc) protein, produced as inclusion bodies in Escherichia coli, was incorporated into phospholipid vesicles consisting of dipalmitoyl phosphatidylcholine and cholesterol. The incorporation of rOpc into the lipid bilayer was demonstrated, and the reconstitution of some native epitopes was tested using a set of monoclonal antibodies. Subcutaneous immunization of Balb/c mice with rOpc-containing vesicles resulted in the generation of a high level of specific antibodies. The elicited antibodies reacted with the native meningococcal protein and showed opsonic activity. PMID- 11483157 TI - Support vector machines for predicting protein structural class. AB - BACKGROUND: We apply a new machine learning method, the so-called Support Vector Machine method, to predict the protein structural class. Support Vector Machine method is performed based on the database derived from SCOP, in which protein domains are classified based on known structures and the evolutionary relationships and the principles that govern their 3-D structure. RESULTS: High rates of both self-consistency and jackknife tests are obtained. The good results indicate that the structural class of a protein is considerably correlated with its amino acid composition. CONCLUSIONS: It is expected that the Support Vector Machine method and the elegant component-coupled method, also named as the covariant discrimination algorithm, if complemented with each other, can provide a powerful computational tool for predicting the structural classes of proteins. PMID- 11483158 TI - Glycogen synthase kinase3 beta phosphorylates serine 33 of p53 and activates p53's transcriptional activity. AB - BACKGROUND: The p53 protein is activated by genotoxic stress, oncogene expression and during senescence, p53 transcriptionally activates genes involved in growth arrest and apoptosis. p53 activation is regulated by post-translational modification, including phosphorylation of the N-terminal transactivation domain. Here, we have examined how Glycogen Synthase Kinase (GSK3), a protein kinase involved in tumorigenesis, differentiation and apoptosis, phosphorylates and regulates p53. RESULTS: The 2 isoforms of GSK3, GSK3alpha and GSK3beta, phosphorylate the sequence Ser-X-X-X-Ser(P) when the C-terminal serine residue is already phosphorylated. Several p53 kinases were examined for their ability to create GSK3 phosphorylation sites on the p53 protein. Our results demonstrate that phosphorylation of serine 37 of p53 by DNA-PK creates a site for GSK3beta phosphorylation at serine 33 in vitro. GSK3alpha did not phosphorylate p53 under any condition. GSK3beta increased the transcriptional activity of the p53 protein in vivo. Mutation of either serine 33 or serine 37 of p53 to alanine blocked the ability of GSK3beta to regulate p53 transcriptional activity. GSK3beta is therefore able to regulate p53 function in vivo. p53's transcriptional activity is commonly increased by DNA damage. However, GSK3beta kinase activity was inhibited in response to DNA damage, suggesting that GSK3beta regulation of p53 is not involved in the p53-DNA damage response. CONCLUSIONS: GSK3beta can regulate p53's transcriptional activity by phosphorylating serine 33. However, GSK3beta does not appear to be part of the p53-DNA damage response pathway. Instead, GSK3beta may provide the link between p53 and non-DNA damage mechanisms for p53 activation. PMID- 11483159 TI - Acquisition of tolerance against oxidative damage in Saccharomyces cerevisiae. AB - BACKGROUND: Living cells constantly sense and adapt to redox shifts by the induction of genes whose products act to maintain the cellular redox environment. In the eukaryote Saccharomyces cerevisiae, while stationary cells possess a degree of constitutive resistance towards oxidants, treatment of exponential phase cultures with sub-lethal stresses can lead to the transient induction of protection against subsequent lethal oxidant conditions. The sensors of oxidative stress and the corresponding transcription factors that activate gene expression under these conditions have not yet been completely identified. RESULTS: We report the role of SOD1, SOD2 and TPS1 genes (which encode the cytoplasmic Cu/Zn superoxide dismutase, the mitochondrial Mn-isoform and trehalose-6-phosphate synthase, respectively) in the development of resistance to oxidative stress. In all experimental conditions, the cultures were divided into two parts, one was immediately submitted to severe stress (namely: exposure to H2O2, heat shock or ethanol stress) while the other was initially adapted to 40 degrees C for 60 min. The deficiency in trehalose synthesis did not impair the acquisition of tolerance to H2O2, but this disaccharide played an essential role in tolerance against heat and ethanol stresses. We also verified that the presence of only one Sodp isoform was sufficient to improve cellular resistance to 5 mM H2O2. On the other hand, while the lack of Sod2p caused high cell sensitivity to ethanol and heat shock, the absence of Sod1p seemed to be beneficial to the process of acquisition of tolerance to these adverse conditions. The increase in oxidation-dependent fluorescence of crude extracts of sod1 mutant cells upon incubation at 40 degrees C was approximately 2-fold higher than in sod2 and control strain extracts. Furthermore, in Western blots, we observed that sod mutants showed a different pattern of Hsp104p and Hsp26p expression also different from that in their control strain. CONCLUSIONS: Trehalose seemed not to be essential in the acquisition of tolerance to H2O2 stress, but its absence was strongly felt under water stress conditions such as heat and alcoholic stresses. On the other hand, Sod1p could be involved in the control of ROS production; these reactive molecules could signal the induction of genes implicated within cell tolerance to heat and ethanol. The effects of this deletion needs further investigation. PMID- 11483160 TI - June 5, 2001: marking 20 years since the recognition of AIDS. PMID- 11483161 TI - Possible linkage of amprenavir with intracranial bleeding in an HIV-infected hemophiliac. AB - The use of protease Inhibitors (PI) has been associated with many adverse effects including increased tendency to bleed, which is particularly problematic in individuals with congenital coagulation disorders. We report the occurrence of spontaneous intracranial bleeding in an human immunodeficiency virus (HIV) infected adolescent with hemophilia A who was receiving amprenavir (APV). The bleeding resolved on discontinuation of APV. This case report highlights a need for awareness of increased bleeding as a potentially serious complication associated with the use of all currently licensed PIs in individuals with hemophilia. PMID- 11483162 TI - Cavitary lung disease in AIDS: etiologies and correlation with immune status. AB - To investigate the etiology and differential features of cavitary lung disease in patients with acquired immune deficiency syndrome (AIDS), chest computed tomography (CT) records from a 2-year period were reviewed to identify all human immunodeficiency virus (HIV)-positive patients with cavitary lung disease. Medical records were reviewed for the documentation of specific causes of lung cavitation and the CD4 count at the time of imaging. Of 25 HIV-positive patients with cavitary lung disease, 20 had specific diagnoses. Infection was the etiology in all the cases. Polymicrobial infection was found in 17 patients (85%) and unimicrobial in 3 (15%). Seventeen patients (85%) had bacterial organisms, 10 of whom had other pathogens as well. Mycobacteria were isolated in 8 patients (40%), fungi in 3 (15%), cytomegalovirus (CMV) in 3 (15%), and Pneumocystis carinii pneumonia (PCP) in 1 (5%). Mediastinal or hilar lymphadenopathy and additional noncavitary ill-defined nodular opacities were found more frequently in patients with mycobacterial pathogens. Mean CD4 count in patients with cavitary disease because of bacterial pathogens alone was significantly higher than in patients with nonbacterial pathogens (alone or combined with bacterial pathogens) (203 vs. 42, p < 0.05). Four patients expired during the diagnostic hospital admission; 2 of them had pulmonary cavitary disease associated with Nocardia asteroides. Cavitary lung disease in patients with AIDS undergoing chest CT should be assumed infectious and is generally polymicrobial. PMID- 11483163 TI - Treatment of HIV-associated nephropathy. AB - HIV-associated nephropathy (HIVAN) is the most common cause of renal failure in patients infected with type 1 human immunodeficiency virus (HIV-1). The renal prognosis for HIVAN is poor and is typically associated with rapid progression to renal death. We report a patient with biopsy-proven HIVAN who was successfully treated with corticosteroids and review the currently available evidence supporting the specific treatments of this condition. A 34-year-old African American male with a 2-year history of uncomplicated HIV disease developed progressive azotemia despite treatment with highly active antiretroviral therapy (HAART). He was treated with an uncomplicated 4-month course of prednisone, which improved his serum creatinine from 2.9 to 1.9 mg/dl and decreased his degree of proteinuria from 8 to 2.1 g/day. Two years post-steroid treatment his renal function remains stable. Increasing evidence supports that both ACE inhibitors and HAART are effective in preventing and in some cases of reversing HIVAN induced renal failure. In selected patients who progress despite these measures, a limited course of corticosteroid may achieve long-standing disease remissions. In general, with adequate supervision, corticosteroid therapy appears to be well tolerated and has an acceptable side effect profile. Although persuasive in view of the abysmal natural history of HIVAN, the currently available studies are subject to major methodological limitations. Appropriate randomized controlled trials are urgently required in order to further examine the efficacy, optimal timing, and potential side effects of these treatments. PMID- 11483164 TI - Nursing home residents with HIV and anemia. AB - This research profiles nursing home residents who have human immunodeficiency virus (HIV) and anemia at the time of admission, utilizing the minimum data set (MDS). In addition, this article compares residents with HIV and anemia to other nursing home residents with HIV. These resident profiles include sociodemographic characteristics, health status measures, and special treatments and procedures received. This study analyzed 1,281 admission assessments for HIV residents with anemia and 3,832 admission assessments for other residents with HIV in the MDS between June 22, 1998 and January 17, 2000. A significantly greater percentage of HIV residents with anemia were female (38.6%) compared to other residents with HIV (27.9% female). Almost two-thirds of HIV residents with anemia and three quarters of other residents with HIV received Medicaid coverage at the time of their admission to the nursing home. Approximately 3 of every 4 residents with HIV and anemia and other residents with HIV were from racial/ethnic minority groups. Significantly greater percentages of residents with HIV and anemia also had dementia, depression, pneumonia, hepatitis, renal failure, anxiety disorder, and cancer than other residents with HIV. These analyses demonstrate that at the time of admission to the nursing home, those residents with HIV and anemia were significantly more likely to have other diseases, infections, and health care conditions than other residents with HIV. In addition, HIV residents with anemia were significantly more likely to receive special treatments and procedures in the nursing home than other residents with HIV. PMID- 11483165 TI - Osteonecrosis and highly active antiretroviral therapy during HIV infection: report of a series and literature review. AB - Osteonecrosis has shown an increased incidence in HIV-infected patients during the last few years, and has been looked as a side effect of highly active antiretroviral therapy, possibly related to hyperlipidemia. Five cases of osteonecrosis were observed among over 1300 patients with HIV disease. According to our experience, novel antiretroviral regimens do not represent a constant risk factor for avascular necrosis of bone during HIV infection. PMID- 11483166 TI - Late diagnosis of HIV infection in women seeking counseling and testing services in Sao Paulo, Brazil. AB - We investigated whether women positive for human immunodeficiency virus (HIV) infection were late in seeking an anonymous HIV counseling and testing service, and the factors associated with a low CD4 count, in Sao Paulo, Brazil. Seventy one consecutive HIV-1-seropositive women were interviewed by means of a structured questionnaire. Blood samples were collected for CD4+ T-lymphocytes count and determination of HIV-1 subtypes. Hepatitis C, syphilis, human T-cell lymphotrophic virus type I (HTLV-I), and HTLV-II infections were assessed by serologic tests. More than 70% of the women had less than 500 CD4+ cells/mm3 (20% below 200). Low CD4 count was significantly associated with sex work history, condom use in the last 6 months, and seropositivity to HTLV-I and syphilis. There was no relation between low CD4 count and HIV-1 subtypes. These results indicate that in Sao Paulo many women are seeking an anonymous testing service late in the course of HIV infection. The main purposes of anonymous HIV testing services - early diagnosis of infection, and counseling to prevent infection - are not being achieved. Another strategy for reducing the interval between infection and diagnosis in women must be addressed. PMID- 11483167 TI - Tenofovir ready for approval. PMID- 11483168 TI - Planned Parenthood has own condom brand. PMID- 11483169 TI - Spatial language and spatial representation: a cross-linguistic comparison. AB - We examined the relationship between spatial language and spatial memory by comparing native English, Japanese, and Korean speakers' naming of spatial locations and their spatial memory for the same set of locations. We focused on two kinds of spatial organization: axial structure of the reference object, and contact/support with respect to its surface. The results of two language (naming) tasks showed similar organization across the three language groups in terms of axial structure, but differences in organization in terms of contact/support. In contrast, the results of two memory tasks were the same across language groups for both axial structure and contact/support. Moreover, the relationship between spatial language and spatial memory in the two sets of tasks did not show a straightforward isomorphism between the two systems. We conclude that spatial language and spatial memory engage the same kinds of spatial properties, suggesting similarity in the foundations of the two systems. However, the two systems appear to be partially independent: the preservation of particular spatial properties was not mandatory across languages, nor across memory tasks, and cross-linguistic differences in spatial language did not lead to differences in the non-linguistic encoding of location. We speculate that the similarity in linguistic and non-linguistic representations of space may emerge as a functional consequence of negotiating the spatial world. PMID- 11483170 TI - Anticipating incoming events: an impaired cognitive process in schizophrenia. AB - Intentions are central to guiding actions to their completion because they generate expectations which precede the realization of a task. This ability to manage time was investigated by using a cognitive task which involves several highly integrated processes: sequential learning, explicit processing, and working memory. In this task, participants are required to explicitly learn a repeating color sequence before receiving an instruction to give an anticipatory motor response concerning the next element. Two types of sequences (temporal and spatial) and three experimental conditions were tested in both a group of normal participants and a group of schizophrenic patients. Schizophrenics were included because their condition is known to alter conscious executive function. Our results showed that schizophrenic patients have a strong deficit in performing anticipation tasks. Although they learned the sequences almost normally, their anticipatory ability was reduced in comparison to normal participants in all the tested conditions. These results expand the notion of a working memory deficit in schizophrenia and bear strong implications for understanding executive disorders observed in such patients. PMID- 11483171 TI - The development of phonological awareness: effects of spoken language experience and orthography. AB - Phonological awareness, the ability to analyze spoken language into small sound units, has been shown to be affected by the individual's early orthographic experience (alphabetic vs. non-alphabetic). Past studies, however, have not differentiated the effect of script alphabeticity from that of spoken language experience, which covaries strongly with the phonological properties of the language. The present study compares younger, pre-reading to older, literate children from different linguistic backgrounds on their phonological awareness. Hong Kong and Guangzhou subjects both spoke Cantonese. The latter subjects had early experience with Pinyin (alphabetic) in addition to their logographic Chinese reading; the former read only logographic Chinese. New Zealand subjects spoke English and read the Roman alphabet. Results showed that: (1) the Hong Kong and Guangzhou pre-readers performed very similarly at all levels of phonological awareness; (2) the New Zealand pre-readers outperformed their Hong Kong and Guangzhou counterparts on onset, rime, and coda analyses; (3) the Guangzhou reading children outperformed their Hong Kong counterparts on onset and coda analyses. Whereas finding (3) reflects an effect of alphabeticity in the first learned script, finding (2) in combination with finding (1) indicates an effect of early spoken language experience independent of orthography. The fact that orthographic and spoken language experience both impact on the development of phonological skills implies a mediating function of phonological awareness in integrating sound information derived from reading and perceiving speech. PMID- 11483172 TI - Effects of semantic context in the naming of pictures and words. AB - Two experiments investigated whether lexical retrieval for speaking can be characterized as a competitive process by assessing the effects of semantic context on picture and word naming in German. In Experiment 1 we demonstrated that pictures are named slower in the context of same-category items than in the context of items from various semantic categories, replicating findings by Kroll and Stewart (Journal of Memory and Language, 33 (1994) 149). In Experiment 2 we used words instead of pictures. Participants either named the words in the context of same- or different-category items, or produced the words together with their corresponding determiner. While in the former condition words were named faster in the context of same-category items than of different-category items, the opposite pattern was obtained for the latter condition. These findings confirm the claim that the interfering effect of semantic context reflects competition in the retrieval of lexical entries in speaking. PMID- 11483173 TI - Wisdom and aging: irrational preferences in college students but not older adults. AB - A decision-maker is "irregular" if she would choose B from [A, B, C] but not from [A, B] (for example, preferring vanilla ice cream in a choice between vanilla and chocolate, but chocolate in a choice among vanilla, chocolate and strawberry). Similarly to previous studies we observed irregular choices by college students faced with hypothetical discount cards for supermarkets. However, older adults showed no such tendency. The same pattern was observed in three separate studies. We interpret the results in terms of a choice strategy by older adults that protects them from excessive spending. PMID- 11483174 TI - Ultraviolet and middle wavelength sensitive cone responses in the electroretinogram (ERG) of normal and Rpe65 -/- mice. AB - Ultra-violet (UV) and middle wavelength sensitive (M) cone responses were identified in the ERG of normal and Rpe65 -/- mice using chromatic flashes and selective chromatic adaptation. In normal mice, the UV-cone response was as large as, or larger, in the presence of a bright yellow adapting light than it is in the presence of a dim white light. The M-cone response became undetectable in the presence of the yellow adapting light. Yellow adapting light initially reduced the UV response, but it recovered in 8-10 min. The M-cone response did not recover. UV-cone responses were undetectable in Rpe65 -/- mice. The M-cone response of young Rpe65 -/- mice was almost as large as in normal mice. A yellow adapting light only diminished this M-cone response. With age, the M-cone response further decreased in Rpe -/- mice. We show a pronounced loss of UV-cone function in Rpe65 -/- mice, which may be related to a defect UV-cones share with rods. The M-cone function is also affected already in young Rpe65 -/- mice. The transient effect of a yellow adapting light on the UV-cone response of normal mice is suggested to be neural, because it disappears during maintained light adaptation. PMID- 11483175 TI - Automatic gain control contrast mechanisms are modulated by attention in humans: evidence from visual evoked potentials. AB - This study investigated the effect of attention on the contrast response curves of steady-state visual evoked potentials (VEPs) to counter-phased sinusoidal gratings. The 1 cyc/deg gratings were modulated either in luminance or chromaticity (equiluminant red-green). The luminance grating counter-phased at 9 Hz (to favour activation of the magno-cellular system), and the chromatic grating at 2.5 Hz (to favour activation of the parvo-cellular system). Attention was directed towards the gratings (displayed in the left visual field) by requiring subjects to detect and respond to randomly occurring changes in contrast. In the control condition, attention towards the grating was minimised by requiring subjects to detect a target letter amongst distracters briefly flashed in the contra-lateral visual field. Attention increased VEP amplitudes for both luminance and chromatic stimuli, more so at high than at low contrasts, increasing the slope of the contrast amplitude curves (over the non-saturating range of contrasts). The estimates of contrast threshold from extrapolation of amplitudes were unaffected by attention. Attention also changed the VEP phases, but only for luminance gratings, where it acted to reduce the magnitude of phase advance with contrast. Attention had no effect on the average phases for chromatic gratings. The results are consistent with the notion that attention acts on cortical gain control mechanisms, which are known to be different for the magno- and parvo-cellular systems. PMID- 11483176 TI - Multiple processes mediate flicker sensitivity. AB - By systematically manipulating the luminance of a flickering spot and the area immediately surrounding it, we investigated why thresholds from flickering stimuli that cause a change in average luminance are elevated relative to those from stimuli with no luminance change. Threshold elevation resulted from local light adaptation and from temporal-frequency-specific interactions between the spot and its surround: at low frequencies, the contrast between the spot and the surround elevated thresholds, whereas at high frequencies, dark adaptation within the surround elevated thresholds. Our findings suggest that two common ways of determining temporal sensitivity may give markedly different outcomes. PMID- 11483177 TI - Are corresponding points fixed? AB - Several investigators have claimed that the retinal coordinates of corresponding points shift with vergence eye movements. Two kinds of shifts have been reported. First, global shifts that increase with retinal eccentricity; such shifts would cause a flattening of the horopter at all viewing distances and would facilitate fusion of flat surfaces. Second, local shifts that are centered on the fovea; such shifts would cause a dimple in the horopter near fixation and would facilitate fusion of points fixated at extreme viewing distances. Nearly all of the empirical evidence supporting shifts of corresponding points comes from horopter measurements and from comparisons of subjective and objective fixation disparity. In both cases, the experimenter must infer the retinal coordinates of corresponding points from external measurements. We describe four factors that could affect this inference: (1) changes in the projection from object to image points that accompany eye rotation and accommodation, (2) fixation errors during the experimental measurements, (3) non-uniform retinal stretching, and (4) changes in the perceived direction of a monocular point when presented adjacent to a binocular point. We conducted two experiments that eliminated or compensated for these potential errors. In the first experiment, observers aligned dichoptic test lines using an apparatus and procedure that eliminated all but the third error. In the second experiment, observers judged the alignment of dichoptic afterimages, and this technique eliminates all the errors. The results from both experiments show that the retinal coordinates of corresponding points do not change with vergence eye movements. We conclude that corresponding points are in fixed retinal positions for observers with normal retinal correspondence. PMID- 11483178 TI - Sensitivity to modulation of color distribution in multicolored textures. AB - We evaluated the discriminability of color distributions in square-element textures. Each texture contained 225 colors, represented by a distribution of color vectors in color space defined by the L-M and S-(L+M) axes. Each color distribution was systematically manipulated by modulating the distribution of the vector lengths sinusoidally as a function of the direction in the color space. The results showed that it is difficult to resolve a color distribution modulated in more than three cycles per 360 degrees in the chromatic direction. The difference in components along the cardinal axes is not a critical factor in the discrimination of the color distribution. An analysis using a line-element model suggested that the discrimination of the color distribution is mediated by multiple chromatic channels that are tuned to a variety of directions in the color space with a half-height-half-bandwidth of about 40 degrees in the chromatic direction. PMID- 11483179 TI - A multiscale spatial filtering account of the Wertheimer-Benary effect and the corrugated Mondrian. AB - Blakeslee and McCourt [Blakeslee, B., & McCourt, M.E. (1997). Similar mechanisms underlie simultaneous brightness contrast and grating induction. Vision Research, 37, 2849-2869] demonstrated that a multiscale array of two-dimensional difference of-Gaussian (DOG) filters provided a simple but powerful model for explaining a number of seemingly complex features of grating induction (GI), while simultaneously encompassing salient features of brightness induction in simultaneous brightness contrast (SBC), brightness assimilation and Hermann Grid stimuli. The DOG model (and isotropic contrast models in general) cannot, however, account for another important group of brightness effects including the White effect [White, M. (1997). A new effect of pattern on perceived lightness. Perception, 8, 413-416] and a variant of SBC [Todorovic, D. (1997). Lightness and junctions. Perception, 26, 379-395]. Blakeslee and McCourt [Blakeslee, B., McCourt, M.E. (1999). A multiscale spatial filtering account of the White effect, simultaneous brightness contrast and grating induction. Vision Research, 39, 4361 4377] developed a modified version of the model, an oriented (ODOG) model, which differed from the DOG model in that the filters were anisotropic and their outputs were pooled nonlinearly. Using this model, they were able to account for both groups of induction effects. The present paper examines two additional sets of brightness illusions that cannot be explained by isotropic contrast models. Psychophysical brightness matching is employed to quantitatively measure the size of the brightness effect for two Wertheimer-Benary stimuli [Benary, W. (1924). Beobachtungen zu einem experiment uber helligkeitskontrast. Psychologische Forschung, 5, 131-142; Todorovic, D. (1997). Lightness and junctions. Perception, 26, 379-395] and for low- and high-contrast versions of corrugated Mondrian stimuli [Adelson, E.H. (1993). Perceptual organization and the jugdement of brightness. Science, 262, 2042-2044; Todorovic, D. (1997). Lightness and junctions. Perception, 26, 379-395]. Brightness matches are obtained on both homogeneous and checkerboard matching backgrounds. The ODOG model qualitatively predicts the appearance of the test patches in the Wertheimer-Benary stimuli and corrugated Mondrian stimuli. In addition, it quantitatively predicts the relative magnitudes of the corrugated Mondrian effects in the various conditions. In general, the psychophysical results and ODOG modeling argue strongly that like SBC, GI, the White effect and Todorovic's SBC demonstration, induced brightness in Wertheimer-Benary stimuli and in the corrugated Mondrian primarily reflects early-stage filtering operations in the visual system. PMID- 11483180 TI - Eye movement control in reading unspaced text: the case of the Japanese script. AB - The present study examines the landing-site distributions of the eyes during natural reading of Japanese script: a script that mixes three different writing systems (Kanji, Hiragana, and Katakana) and that misses regular spacing between words. The results show a clear preference of the eyes to land at the beginning rather than the center of the word. In addition, it was found that the eyes land on Kanji characters more frequently than on Hiragana or Katakana characters. Further analysis for two- and three-character words indicated that the eye's landing-site distribution differs depending on type of the characters in the word: the eyes prefer to land at the word beginning only when the initial character of the word is a Kanji character. For pure Hiragana words, the proportion of initial fixations did not differ between character positions. Thus, as already indicated by Kambe (National Institute of Japanese Language Report 85 (1986) 29), the visual distinctiveness of the three Japanese scripts plays a role in guiding eye movements in reading Japanese. PMID- 11483181 TI - The effect of reading and near-work on the development of myopia in emmetropic boys: a prospective, controlled, three-year follow-up study. AB - This study aimed to investigate the effect of reading and near work on myopic development in emmetropic boys in school age. It involved totally 114 children in two groups. Right eyes of 67 randomly selected students (mean age=12.93) with mean 6 h of reading and near work (Group 1) were compared with the right eyes of 47 apprentices (mean age=12.96) working as skilled laborers (Group 2). Cycloplegic refraction, keratometric readings and biometric measurements including anterior chamber depth (ACD), lens thickness (LT), vitreous chamber depth (VCD) and axial length (AL) were performed for 3 years at 18 month intervals. Two analyses were conducted: (1) for subjects in both groups with baseline refractive error from +0.50 to -0.50 D; (2) for all subjects in both groups with baseline refractive error from +1.00 to -1.00 D. For subjects with baseline refractive error of +/-0.50 D, myopic shift was present in 20 of 41 (48.8%) in group 1 and in seven of 37 (18.9%) in group 2 at the end of the study. The magnitude of the myopic shift was 0.56 and 0.07 D in group 1 and 2, respectively. For subjects with a baseline refractive error of +/-1.00 D, myopic progression was present in 40 of 67 (59.7%) in group 1 and in 10 of 47 (21.3%) in group 2 at the last readings. In this larger refractive range, the magnitude of the myopic shift was 0.61 and 0.12 D in group 1 and 2, respectively. The mean ACD, VCD and AL were significantly higher in the last readings after 36 months than in the first readings (for each, P=0.0001) in group 1. There was no statistically significant difference between two measurements of these parameters in group 2. The final keratometric dioptric readings were lower than the first values (for each, P=0.0001) in both groups at the end of the study. This prospective and controlled study suggested that reading and near work, important environmental factors, might cause refractive myopic shifts in emmetropic students. The myopic shift was primarily related to significant increases in ACD, VCD and AL in this young age group. PMID- 11483182 TI - Neural dynamics of motion integration and segmentation within and across apertures. AB - A neural model is developed of how motion integration and segmentation processes, both within and across apertures, compute global motion percepts. Figure-ground properties, such as occlusion, influence which motion signals determine the percept. For visible apertures, a line's terminators do not specify true line motion. For invisible apertures, a line's intrinsic terminators create veridical feature-tracking signals. Sparse feature-tracking signals can be amplified before they propagate across position and are integrated with ambiguous motion signals within line interiors. This integration process determines the global percept. It is the result of several processing stages: directional transient cells respond to image transients and input to a directional short-range filter that selectively boosts feature-tracking signals with the help of competitive signals. Then, a long-range filter inputs to directional cells that pool signals over multiple orientations, opposite contrast polarities, and depths. This all happens no later than cortical area MT. The directional cells activate a directional grouping network, proposed to occur within cortical area MST, within which directions compete to determine a local winner. Enhanced feature-tracking signals typically win over ambiguous motion signals. Model MST cells that encode the winning direction feed back to model MT cells, where they boost directionally consistent cell activities and suppress inconsistent activities over the spatial region to which they project. This feedback accomplishes directional and depthful motion capture within that region. Model simulations include the barberpole illusion, motion capture, the spotted barberpole, the triple barberpole, the occluded translating square illusion, motion transparency and the chopsticks illusion. Qualitative explanations of illusory contours from translating terminators and plaid adaptation are also given. PMID- 11483184 TI - Anterior approaches to cervical spondylosis and ossification of the posterior longitudinal ligament: review of operative technique and assessment of 65 multilevel circumferential procedures. AB - BACKGROUND: Multilevel anterior cervical corpectomy with fusion (ACF) offers direct resection of spondylostenosis and ossification of the posterior longitudinal ligament (OPLL) with immediate stabilization. Ideal candidates for multilevel ACF include younger patients (<65 years of age), or older individuals (>65 years of age) with loss or reversal of the cervical lordosis (kyphosis). METHODS: Sixty-five patients, averaging 56 years of age and including 40 males and 25 females, with multilevel MR- and CT-documented spondylostenosis and OPLL were studied. Preoperatively, patients exhibited moderate to severe myelopathy (average Nurick grade 3.8), and were managed with 2- to 4-level ACF with posterior wiring and fusion (PWF) procedures with halo application. The first 22 patients had no plate instrumentation, the next 22 had constrained (Orion) plates applied followed sequentially by the application of 13 semi-constrained (Atlantis) plates, and finally, 8 dynamic (ABC Aesculap) plates. RESULTS: Patients improved an average of three postoperative Nurick grades. None exhibited new cord injuries, whereas three had transient C5 root paresis. Graft/plate or vertebral fracture with extrusion were observed in 3/22 nonplated patients, 2/22 constrained-plated patients, 3/13 semi-constrained-plated patients, and 0/8 dynamic-plated patients. Fusion was documented on dynamic radiographs and 2D-CT or 3D-CT studies obtained 3 and 6 months postoperatively, or later where indicated. CONCLUSIONS: Multilevel ACF/PWF offers direct resection of spondylostenosis and OPLL with immediate maximal stabilization. Thus far, no graft/plate or vertebral body fracture or extrusions have been seen with dynamic plates, whereas the absence of plating and constrained and semi-constrained plating systems have failed. PMID- 11483185 TI - Long-term results of stereotactic gamma radiosurgery of meningiomas. AB - BACKGROUND: The early effects at a mean of 30 months and long-term results at 7 years after gamma radiosurgery for meningiomas were evaluated. METHODS: Changes in tumor size were evaluated every 3 to 6 months after treatment using a five point grading system, as well as changes in neurological signs and general status. RESULTS: Early effects in 87 cases of benign meningioma showed a minimal size reduction of 16.1% and a response rate of 8.0%, but a higher control rate of 93%. The cavernous sinus meningioma showed a size reduction of 23.2%, a response rate of 11.1%, and control of 100%. A greater size reduction of 24.8% and response rate of 33.3%, but a lower control rate of 75% were obtained in 12 cases of malignant meningioma. Side effects were found in 12 cases (13.8%): radiation induced edema in 9, hearing disturbance in 2, and visual deterioration in 1. Long term results for 54 of 87 patients with benign tumors showed that response increased from 8% to 42.6% but control decreased slightly due to increased disease progression. CONCLUSION: Gamma radiosurgery is effective and safe for meningiomas to control residual or recurrent tumors after surgery and initial tumors, with acceptable side effects and rate of tumor progression. PMID- 11483187 TI - Primum non nocere: Multimodality management strategies when multiple mass lesions strike a single patient. AB - BACKGROUND: The management of multiple symptomatic intracranial pathological processes in a single patient presents a rare and challenging problem for the neurosurgeon and the patient. Neurosurgeons must utilize a full spectrum of neurosurgical options to achieve the best patient outcome. CASE DESCRIPTION: We present a unique case of a 63-year-old woman who presented with a large convexity meningioma causing headaches, an acoustic neuroma causing deafness and imbalance and a suprasellar arachnoid cyst compromising the visual fields. Therapeutic intervention was staged based on the primum non nocere concept. First, the patient underwent stereotactic intracavitary cyst irradiation using colloidal 32P. Secondly, microsurgical resection of the convexity meningioma was performed. Finally, Gamma Knife radiosurgery of the acoustic neuroma was performed. One year after multimodality management, the patient was neurologically improved. There was no evidence of meningioma or cyst recurrence and the growth of the acoustic neuroma was arrested. CONCLUSION: This case demonstrates the value of multi modality treatment of neurosurgical pathology, utilizing minimally invasive techniques when possible. PMID- 11483188 TI - Treatment of deep cerebral venous thrombosis by local infusion of tissue plasminogen activator. AB - BACKGROUND: Treatment of extensive intracranial venous sinus thrombosis with thrombolytic drugs is described, although the indications for and most efficacious technique for achieving thrombolysis remain uncertain. We report the successful lysis of superficial and deep venous system thrombosis by infusion of recombinant human tissue-type plasminogen activator (rt-PA) into the anterior superior sagittal sinus. CASE DESCRIPTION: A 34-year-old man presented with headaches followed by decreased level of consciousness and left hemiplegia. Angiography showed thrombosis of the superior sagittal and both transverse and straight sinuses with extension into the internal cerebral veins. The superior sagittal sinus was catheterized via a transfemoral route and rt-PA, 25 mg, was infused. There was no significant change in the thrombosis. The catheter was left in place and rt-PA was infused at 1 mg/minute for 19 hours. Repeat angiography showed resolution of the thrombosis. The patient was placed on heparin and then coumadin. He recovered completely. CONCLUSIONS: This report suggests that superselective infusion of thrombolytics into thrombosed intracranial venous sinuses can lyse intracranial venous sinus thrombosis. The thrombolytic agent must be infused for hours. The apparent successful lysis of clot in the deep venous system when infusion was into the superior sagittal sinus might be related to diffusion of rt-PA throughout the intracranial venous system or to improved venous outflow caused by lysis of clot in superficial dural sinuses. PMID- 11483191 TI - Endovascular repair of radiation-induced bilateral common carotid artery stenosis and pseudoaneurysms: a case report. AB - BACKGROUND: Radiation-induced damage to small and medium-sized vessels has been observed in both animals and humans. Changes may appear in the immediate postradiation period or many years later. In this case, we report an unusual presentation of bilateral radiation-induced carotid artery stenoses associated with pseudoaneurysms, and a previously unreported application of a recently established treatment. CASE DESCRIPTION: A 72-year-old African-American male presented with recurrent right hemispheric transient ischemic attacks (TIA) and neck pain. Thirteen years previously, the patient had received radiation therapy for laryngeal carcinoma. Diagnostic carotid angiography demonstrated moderate radiation-induced bilateral carotid artery stenosis and associated common carotid pseudoaneurysms. The patient was treated with bilateral endovascular stents and electrolytically detachable coils in staged procedures. At his most recent follow up, there is no evidence of re-stenosis and the patient remains asymptomatic. CONCLUSIONS: This case illustrates a novel and successful treatment for the endovascular repair of post-radiation bilateral carotid artery stenosis and pseudoaneurysms. In our practice, we have seen three such cases of radiation induced vasculopathy. Therefore, patients with a previous history of radiation therapy for head and neck neoplasms merit cautious monitoring and judicious use of stents and secondary coils, when necessary. PMID- 11483194 TI - Transarterial intravenous coil embolization of dural arteriovenous fistula involving the superior sagittal sinus. AB - BACKGROUND: We report a rare case of traumatic dural arteriovenous fistula involving the superior sagittal sinus successfully treated by transarterial intravenous coil embolization. CASE PRESENTATION: A 38-year-old woman presented with tension headache. She had a past history of severe head injury at the age of three. Computed tomography scanning showed a heterogenous low-density area in the right frontal lobe, and magnetic resonance imaging demonstrated abnormal vascular structures in the same area. Angiography revealed a dural arteriovenous fistula involving the lateral wall of the fully patent superior sagittal sinus. The fistula was fed by scalp, meningeal, and cortical arteries, and drained into a cortical vein leading to the superior sagittal sinus. Femoral transarterial intravenous embolization with microcoils completely occluded the dural arteriovenous fistula. CONCLUSION: Severe head injury may lead to asymptomatic dural arteriovenous fistulas after a long time. Transarterial intravenous coil embolization can be effective in the treatment of dural arteriovenous fistulas involving the superior sagittal sinus. PMID- 11483196 TI - The usefulness of 3D MR angiography in surgery for ruptured cerebral aneurysms. AB - BACKGROUND: We have used magnetic resonance angiography (MRA) in screening for unruptured cerebral aneurysms since 1993. The development of high-resolution magnetic resonance (MR) imaging has led to a remarkable improvement in image quality. Three-dimensional (3D) MRA can be used for surgical simulation. Here, we report on the usefulness of and problems associated with 3D MRA for the surgery of ruptured cerebral aneurysms. METHODS: Between June 1998 and June 2000, 106 patients with SAH diagnosed by 3D MRA underwent surgery. We compared 3D MRA images with operative findings and investigated the usefulness of this assessment tool. RESULTS: In 48 of 106 cases (45.3%), we were able to perform surgery based on 3D MRA alone. By using the 3D images, we could easily detect the relative location of the aneurysm, its neck and the surrounding arteries. The remaining cases required further examinations because of uncertainty of diagnosis or insufficient information. CONCLUSION: 3D MRA is a safe and useful procedure for the diagnosis and surgery of ruptured cerebral aneurysms. However, in approximately half of all cases, 3D computed tomographic angiography (CTA) or digital subtraction angiography (DSA) is required in addition for the planning of surgery. It is important to use 3D MRA for surgery only after taking sufficient consideration of certain limitations peculiar to MRA. PMID- 11483199 TI - Pineal apoplexy due to massive hemorrhage associated with cavernous angioma: case report. AB - BACKGROUND: Pineal apoplexy is a rare apoplectic event in the pineal region with various possible causes. We report a case of massive hemorrhage in the pineal region associated with a cavernous angioma, and discuss the pathogenesis of pineal apoplexy. CASE DESCRIPTION: An 11-year-old girl presented with nausea and vomiting persisting for 1 week. Neuroimaging revealed enlarged ventricles and a 4 cm round mass in the pineal region consistent with hematoma. Two weeks after cerebrospinal fluid drainage, the mass was totally removed via the occipital transtentorial approach. The postoperative course of the patient was uneventful. Magnetic resonance imaging after the operation confirmed two paraventricular lesions with surrounding hemosiderin rings, characteristic of cavernous angioma. Histologic examination showed large hematomas and numerous vascular spaces lined by a single layer of endothelium. The diagnosis was pineal apoplexy associated with cavernous angioma, based on the histologic and radiological findings. CONCLUSIONS: Pineal cavernous angiomas are extremely rare, with only fourteen cases reported previously. We recommend total removal of the causative lesion in cases of pineal apoplexy to prevent repeated bleeding or life-threatening massive hemorrhage. PMID- 11483200 TI - Cavernous sinus tuberculoma: diagnostic difficulties in a personal case. AB - BACKGROUND: Tuberculoma of the brain is rare, and its location in the cavernous sinus is exceptional. Many factors make the diagnosis difficult, necessitating a surgical procedure. CASE DESCRIPTION: A 44-year-old woman complained of persistent headaches and diplopia. The examination revealed a right cavernous sinus syndrome. Radiologic investigation showed a growing process in the right cavernous sinus, overlapping the sella turcica. The diagnoses of metastasis, tuberculoma, and sarcoidosis were considered, but none of them could be confirmed. A biopsy of the lesion via a subtemporal approach revealed a tuberculoma. Antituberculous treatment led to complete resolution of her symptoms. CONCLUSION: The diagnosis of a cavernous sinus tuberculoma is difficult; it is confirmed by biopsy. However, a trial of medical treatment, if the results are favorable, may obviate the need for a surgical procedure. PMID- 11483202 TI - Free hand aspiration for large periventricular hemorrhage. Case report. AB - BACKGROUND: At present, there are several therapeutic options, including craniotomy and stereotactic aspiration, for large intracerebral hemorrhage perforating into the lateral ventricle. In the cases with Glasgow Coma Scale (GCS) scores under 6 with anisocoria, external ventricular drainage would be the first choice [2-4]. We have also performed anterior ventricular horn puncture in a standard manner. The target was the foramen of Monro, at a depth of 5.5 cm from the inner table of the skull. The point of insertion was located just anterior to the coronal suture, approximately 10 cm above the nasion, and 3 cm from the midline [1]. However, we noticed that the insertion of a catheter into the periventricular hematoma adjacent to the lateral ventricle was made easier by tilting the catheter 30 degrees laterally as in the first case (Figure 1). METHODS: In our method, inclining the catheter by 30 degrees laterally, we used a silicone tube 3.5 mm in internal diameter (Silascon ventricle drainage tube, Kaneka Medix Corp., Osaka, Japan) and then replaced it with another Silascon tube with a 2.5 mm-internal diameter. From January 1999 through December 2000, eleven patients who all had GCS scores under 6 and anisocoria preoperatively were treated by this method. The series included two patients who were undergoing hemodialysis because of renal failure, two with bleeding tendency because of liver dysfunction, and one with heart failure. RESULTS: The insertion itself caused no complications. Cerebrospinal fluid was drained smoothly after removal of hematoma because the hematoma cavity connecting with the lateral ventricle was opened. Two typical cases using this technique are shown (Figures 1 and 2). All patients recovered favorable consciousness postoperatively compared with the preoperative state but hemiparesis remained. Postsurgical follow up at 3 months revealed the outcomes evaluated by Glasgow Outcome Scale (GOS) were moderate disability in 5 patients and severe disability in 6 patients . CONCLUSION: This direct aspiration and drainage of a large intracerebral hematoma that ruptures into the lateral ventricle is superior to simple ventricular drainage in regard to the removal of the hematoma clot. This technique would be the choice especially in patients with serious complications such as cardiac disease and renal failure. PMID- 11483203 TI - Primary amyloidoma of the thoracic spine presenting with acute paraplegia. AB - BACKGROUND: Primary solitary amyloidoma of the spine is a rare disease characterized by localized deposition of amyloid. To the best of our knowledge, there have been only 14 cases previously reported in the literature. Patients with focal spinal amyloidoma usually have relatively long symptomatic periods preoperatively, ranging from 3 weeks to 6 years (mean: 12 months). Only two reported patients had acute paraplegia. We add a third case of a thoracic spine amyloidoma presenting with acute paraplegia. CASE DESCRIPTION: A 65-year-old man presented with a three-day history of progressive paraplegia and urinary retention. He was found to have severe cord compression at T2 on magnetic resonance imaging. He underwent emergent decompressive laminectomy with instrumentation for spinal stabilization. Histopathology revealed abundant amyloid deposits. A systemic work-up was negative for amyloidosis. The patient showed marked neurological improvement with residual mild spastic gait after 1 year. CONCLUSIONS: Primary spinal amyloidoma with acute paraplegia is rare. One stage surgery combining prompt decompression and stabilization of the spinal column is mandatory in cases of spinal amyloidoma with acute myelopathy, because primary solitary amyloidoma carries a good prognosis. PMID- 11483205 TI - The inter-heavy chain disulfide bonds of IgG4 are in equilibrium with intra-chain disulfide bonds. AB - Unlike other immunoglobulin G (IgG) subclasses, IgG4 antibodies in plasma have been reported to be functionally monovalent. In a previous paper, we showed that the apparent monovalency of circulating IgG4 antibodies is caused by asymmetry of plasma IgG4-a large fraction has two antigen-binding sites resulting in bispecificity. We postulated that the generation of bispecific antibodies was caused by a post-secretion mechanism, involving the exchange of IgG4 half molecules (i.e. one heavy and one light chain). This hypothesis was based on the observed instability of the inter-heavy chain disulfide bonds of IgG4. To investigate this instability, we constructed IgG4 mutants and analyzed the covalent interaction between the heavy chains by sodium dodecyl sulfate-poly acrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions. The mutation to serine of one of the hinge cysteines involved in the inter-heavy chain bond formation, Cys226, resulted in a more stable rather than a more labile inter-heavy chain linkage. Moreover, we confirmed that mutating the IgG4 hinge sequence Cys-Pro-Ser-Cys to the IgG1 hinge sequence Cys-Pro-Pro-Cys also markedly stabilizes the covalent interaction between the heavy-chains. These two observations suggested an explanation for the observed instability of the inter heavy chain disulfide bonds: the formation of an alternative, intra-chain cystine. Obviously, this intra-chain cystine cannot be formed in the mutant where Cys226 is replaced by Ser, and cannot easily be formed in the mutant with the IgG1 hinge sequence (Cys-Pro-Pro-Cys) due to the restricted torsional freedom of prolines. We, therefore, postulate that the lack of a covalent heavy-chain interaction in a subpopulation of IgG4 reflects an equilibrium between inter- and intra-chain cystines. Based upon the published structure of the IgG4-related hinge-deleted IgG1 myeloma protein Mcg, we propose a model for the two forms of IgG4 and for the half-molecule exchange reaction, which might result in the formation of bispecific IgG4 antibodies. PMID- 11483206 TI - Germline diversity of the expressed BALB/c VhJ558 gene family. AB - Although the mouse immunoglobulin heavy chain (Igh) locus contains 15 heavy chain V (Vh) gene families, at least half of the Vh gene segments are members of the VhJ558 family. This large Vh gene family represents the least characterized germline coding regions of any of the mouse antigen receptor loci and the contribution of individual VhJ558 genes to the preimmune repertoire is poorly understood. In fact, relatively few germline VhJ558 sequences have been reported for BALB/c, the foundation strain for mouse immunoglobulin genetics and the prototypic strain of the Igh(a) haplotype. Here we present a database consisting of 66 sequences estimated to represent one-half of the total number of functional BALB/c VhJ558 genes. Our results indicate that a subset of the VhJ558 genes is highly expressed in the preimmune repertoire, with just nine Vh sequences accounting for nearly 50% of the VhJ558 heavy chains expressed by splenic B cells. We show that this disparity in the expressed Vh gene repertoire is not due to the position of the Vh genes relative to the Dh cluster or to multiple germline copies of the highly expressed VhJ558 genes. Together, these data constitute the first detailed analysis of functional BALB/c VhJ558 genes, demonstrate a striking bias in the use of particular VhJ558 genes in the preimmune repertoire, and provide sufficient information to study the regulation of the Dh-distal region of the Igh-V locus at the level of individual genes. PMID- 11483207 TI - Comparative characteristics of mu chain and alpha chain transcripts expressed by individual tonsil plasma cells. AB - Plasma cells (PCs) are one of the two major cell types generated during germinal center reactions. To test the hypothesis that PCs express a unique repertoire of immunoglobulin (Ig) genes resulting from intensive antigenic stimulation and selection, the mutational pattern and distribution of V(H) gene segments within 178 transcripts amplified from individual IgM and IgA secreting tonsil PCs were analyzed. The results demonstrated that both mu and alpha transcripts expressed repertoires with limited diversity. Moreover, both mu and alpha transcripts were heavily mutated, with a significantly increased mutational frequency noted for alpha compared to mu transcripts (5.0 x 10(-2) vs 1.8 x 10(-2), P<0.001). In addition, both mu and alpha transcripts showed significantly greater targeting of mutations to RGYW motifs (purine/guanine/pyrimidine/A or T) compared to memory B cells. Finally, clonally expanded cells were detected in alpha but not mu PC compartments. These results indicate that antigen driven stimulation and selection shape the entire expressed PC repertoire, but the impact is greater in alpha expressing PCs. PMID- 11483208 TI - Resolution of rabbit polyclonal anti-fluorescein Fab (IgG) fragments into subpopulations differing in affinity and spectral properties of bound ligand. AB - Fab fragments derived from ten different IgG populations of hyperimmune rabbit polyclonal anti-fluorescein antibodies were further resolved into subfractions based on differences in time-dependent dissociation from an FITC-adsorbent in the presence of 0.1 M fluorescein at 4 degrees C. Fab fragments separated into subpopulations based on specific dissociation times of 0.1 day, 1.0 day, 10 days and 100 days from the adsorbent. Finally, after the 100 days elution step incubation with 6.0 M guanidine-HCl was included to determine total protein concentration of specific anti-fluorescein Fab fragments. Yields of specifically eluted Fab fragments ranged from 12.7 to 84.1% of the total Fab population originally incubated with the adsorbent. All Fab polyclonal populations and subpopulations analyzed quenched the fluorescence of the bound ligand by 90% or greater. None of the plots of protein concentration versus percent yield of the total specific antibody obtained for each of the five resolved fractions constituting a specific polyclonal population conformed to Gaussian distributions. All resolved Fab subpopulations retained bound fluorescein ligand that exhibited significant bathochromic shifts in absorbancy. Based on the extent of the red-shift the antibodies segregated into one of two general spectral families showing either a peak shift to 505-507 nm or to 518-520 nm. The red shift to 518-520 nm appeared unique to rabbit anti-fluorescein antibodies, since corresponding large shifts have not been observed with antibodies derived from other species (e.g. mouse, rat, chicken, etc.). K(d) values determined for the resolved fractions confirmed a continuous progression in affinity from the 0.1day through the 100 days elution. Preliminary isoelectric focusing analyses revealed progressive selection for relatively more homogeneous fractions, especially in the 100 days resolved fraction. PMID- 11483209 TI - MHC class II beta-chain and alphaIIbbeta3 integrin are expressed on T-cell progenitors in embryonic bone marrow. AB - The RR5 monoclonal antibody (mAb) was obtained after immunization of mice with hemopoietic cells from chicken embryos. The cDNA encoding the protein recognized by RR5 was cloned using COS-7 cells transfected with an embryonic bone marrow (BM) cDNA library. The epitope recognized by the RR5 mAb was located on the non polymorphic MHC class II beta-chain molecule. In the embryonic BM, RR5 labeled 50% of the c-kit expressing cells. Previous experiments have shown that the T cell progenitors are present in the MHC class II(+)/c-kit(+) BM population along with myeloid progenitors and that T-cell and myeloid progenitors also express the integrin alphaIIbbeta3. In this study, using intrathymic cell transfer experiments in chicks, we have tested the T-cell differentiation potential of MHC class II/alphaIIbbeta3 double positive cells. It proved to be similar to that of the c-kit/MHC class II positive cells. However, injection of triple positive cells resulted in a selection of cells with an increased T-cell potential. Most of the MHC class II positive cells which do not express c-kit are prone to apoptosis, indicating that these progenitors might need a survival signal via c kit. Interestingly, the MHC class II positive progenitors lose this expression after intrathymic transfer. Taken together our data suggest that the presence of the MHC class II beta-chain molecule on the surface of BM progenitor cells could be implicated in differentiation toward myeloid and lymphoid lineages. PMID- 11483210 TI - Normal TCRbeta transcription and recombination in the absence of the Jbeta2 Cbeta2 intronic cis element. AB - The developmental regulation of antigen receptor gene transcription and recombination are mediated by cis regulatory elements. At the T cell receptor beta chain locus (TCRbeta), two DNase I hypersensitive sites within the Jbeta2 Cbeta2 intron contained binding sites for NF-kappaB and additional nuclear factors and were postulated to be involved in controlling TCRbeta transcription and V(D)J recombination. To test this possibility, we deleted these elements from the mouse genome by homologous recombination and assayed the effect on transcription of both the germline and rearranged TCRbeta locus, and on TCRbeta rearrangement in T and B lymphocytes. We found that TCRbeta transcription and V(D)J recombination and T cell development were normal in these mutant mice. Therefore, the Jbeta2-Cbeta2 intronic elements are dispensable for TCRbeta assembly and function. PMID- 11483211 TI - An alternatively spliced long form of Fas apoptosis inhibitory molecule (FAIM) with tissue-specific expression in the brain. AB - The gene encoding Fas apoptosis inhibitory molecule (FAIM) was cloned by differential display using RNA obtained from Fas-resistant and Fas-sensitive primary murine B lymphocytes. FAIM is highly evolutionarily conserved and broadly expressed, suggesting that its gene product plays a key role in cellular physiology. Here we report the identification of a new, longer form of FAIM (FAIM L) and characterization of the genomic locus that clarifies its origin. The murine FAIM gene is located at chromosome 9f1, a region syntenic to the corresponding location of the human FAIM gene. The gene consists of six exons and contains putative translation initiation sites within exons II and III. The long form of FAIM is generated by all six exons, whereas the originally cloned form of FAIM, now termed FAIM-Short (FAIM-S) is generated from five exons by alternative splicing. FAIM-L is dominantly expressed in the brain whereas FAIM-S is widely expressed in many tissues. PMID- 11483212 TI - Molecular characterization of Newcastle disease viruses isolated from recent outbreaks in Taiwan. AB - A reverse transcription-polymerase chain reaction (RT-PCR) was described which amplified a portion of the F and HN genes of Newcastle disease virus (NDV) isolated from recent outbreaks in Taiwan. The F protein plays an important role in determining the virulence of NDV strains. Sequencing of a region specifying the F protein cleavage site was therefore undertaken and this verified the correlation between deduced amino sequences and pathogenicity. Analysis of the deduced amino acid sequences of the F protein cleavage site showed that all recent Taiwanese isolates in 1999 were velogenic viruses. All the virulent viruses have the amino acid sequence 112RRQKR116 for the C-terminus of the F2 protein and phenylanine (F) at the N-terminus of the F1 protein, residue 117. A phylogenetic tree based on the nucleotide sequences of the F gene revealed that recent Taiwanese NDV isolates responsible for recent outbreaks were classified into two distant genotypes (VI and VII). Genotype VI virus is the first finding in Taiwan and has a highly genetic similarity to European isolates, suggesting that they might have originated from a common ancestor. PMID- 11483213 TI - Identification and subtyping of avian influenza viruses by reverse transcription PCR. AB - Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1-H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates. PMID- 11483214 TI - Comparative evaluation of five serological methods and RT-PCR assay for the detection of IPNV in fish. AB - In the present study, six diagnostic methods for the detection of infectious pancreatic necrosis virus (IPNV) (indirect immunofluorescence, flow cytometry, immunoperoxidase, immunodot blot, immunostaphylococcus-protein A, and RT-PCR) have been comparatively evaluated using the seroneutralization as the reference assay, and 83 Spanish isolates and 3 reference strains. The most reliable methods were flow cytometry and RT-PCR which could detect virus at titers of 1x10(2) and 1x10(3) TCID50/ml, respectively. At a multiplicity of infection of 50, both assays allowed the earliest detection of IPNV at 4 h post-inoculation. Indirect immunofluorescence and immunoperoxidase assays required at least 6 h post inoculation to detect viral antigens. The immunodot blot assay possesses low sensitivity and the immunostaphylococcus-protein A test cannot be applied for routine examination of IPNV. Positive reactions were obtained in 100% of the samples tested by seroneutralization and RT-PCR, 90.4% by the flow cytometry, 80.7% by the indirect immunofluorescence assay, 67.5% by the immunoperoxidase, 62.6% by the immunodot blot, and only 27.7% by immunostaphylococcus-protein A test. Therefore, RT-PCR and flow cytometry were the most appropriate and sensitive methods for the routine detection of IPNV from affected fish. PMID- 11483215 TI - A solid-phase competition ELISA for measuring antibody to foot-and-mouth disease virus. AB - A solid-phase competition ELISA has been developed to measure antibodies to foot and-mouth disease (FMD) virus and has been validated using an extensive range of sera from cattle. The assay uses polyclonal antisera and inactivated purified 146S antigens of FMD virus and was compared with the liquid-phase blocking ELISA and the virus neutralisation test on a range of serum sets. When examining test sera at a 1:5 dilution with a cut-off point of 30% inhibition of reaction, the solid-phase competition ELISA was as sensitive as the liquid-phase blocking ELISA for sera from infected or vaccinated animals. The limit of detection of the solid phase ELISA was similar to that of the liquid-phase assay and both tests had lower limit of detection (i.e. were able to detect lower amounts of antibody) than the virus neutralisation test. The specificity of the solid-phase ELISA was considerably higher than that of the liquid-phase blocking ELISA and almost equivalent to that of the virus neutralisation test. The assay thus retains the sensitivity of the liquid-phase blocking ELISA whilst being easier to use, more robust and specific, and therefore offers an improvement for FMD virus antibody detection. PMID- 11483216 TI - Immunocapture-RT-PCR assay for detection and molecular epidemiology studies of Rabbit Haemorrhagic Disease and European Brown Hare Syndrome viruses. AB - Rabbit Haemorrhagic Disease Virus and European Brown Hare Syndrome Virus are two members of the genus Lagovirus in the family Caliciviridae. They are the causative agents of highly contagious and fatal diseases of rabbits and hares respectively. We adjusted one assay for the detection and the genomic characterisation of each virus, based on viral purification by immunocapture and genomic amplification by reverse transcription-polymerase chain reaction (IC-RT PCR). It is carried out directly with the liver exudate obtained after thawing and suppresses the viral nucleic acid preparation step. This assay combines the advantages of an ELISA test (rapidity) because immunocapture and the RT reaction were carried out in the same microtitre plate, and the advantages of PCR (sensitivity). The procedure described allows the processing of large numbers of samples and is suitable for phylogenetic studies of lagomorphs caliciviruses. In addition, it was compared with sandwich-ELISA used for Rabbit Haemorrhagic Disease or European Brown Hare Syndrome diagnosis. A good correlation was found between ELISA and IC-RT-PCR results for Rabbit Haemorrhagic Disease diagnosis, whereas for European Brown Hare Syndrome diagnosis, the results confirmed the higher sensitivity of the molecular method. PMID- 11483217 TI - Direct diagnosis of human respiratory coronaviruses 229E and OC43 by the polymerase chain reaction. AB - An RT-PCR-hybridization was developed that amplified genetic material from the M protein gene of HCoV-229E and HCoV-OC43. The analytic sensitivity of these original primers were compared with primers defined in the N gene and described previously. The results show that 0.05 TCID50 of HCoV-229E and 0.01 TCID50 of HCoV-OC43 can be detected by this molecular method using the original method. Detection of HCoV-229E and HCoV-OC43 in clinical specimens is possible using this method: 348 respiratory specimens (202 sputum and 146 nasal aspirates) were tested with this RT-PCR-hybridization and 12 human coronavirus are detected (3%). The method could provide a useful tool for demonstrating the role of human coronavirus in infections of the respiratory tract. PMID- 11483218 TI - Comparison of RNA and cDNA transfection methods for rescue of infectious bursal disease virus. AB - Specific alterations in the genetic material of RNA viruses rely on a technique known as reverse genetics. Transfection of cells with the altered generic material is a critical step of this procedure. In this report we have compared RNA and cDNA transfection methods for the efficiency of transient protein expression and rescue of (recombinant) infectious bursal disease virus (IBDV). Quantitative expression analysis of the secreted alkaline phosphatase reporter protein, and qualitative expression levels of an IBDV protein showed both that cDNA transfection results in a much higher level of protein expression than RNA transfection. Because the rescue of a crippled variant of IBDV was achieved consistently using the cDNA transfection method, but failed when we used the RNA transfection method, we favor the cDNA transfection method for the rescue of (recombinant) IBDV from cloned cDNA. PMID- 11483219 TI - 'RETCIF': a rapid, sensitive method for detection of viruses, applicable for large numbers of clinical samples. AB - Rapid detection of viruses in clinical samples is important for continuing appropriate antiviral treatment and discontinuing unnecessary antibacterial treatment, as well as for excluding viral pathogens. Yet detection of viral agents may require numerous susceptible cell lines. Even with the shell vial culture method, it is cumbersome for handling large volumes of specimens. A procedure has been developed, which is time and cost-saving and uses specific cell lines in a 96-well microtitre plate and monoclonal antibodies (RETCIF-rapid enhanced tissue culture immunofluorescence). Each clinical sample was inoculated into 12 different wells with five different cell lines. Enhancement was achieved by sonication, centrifugation and hormonal supplementation to the medium used. Cytomegalovirus (CMV), herpes simplex virus (HSV) and respiratory viruses were detected by monoclonal antibodies on day 2, whilst varicella zoster virus (VZV) and enteroviruses were detected on days 5 and 7, respectively. During July December 1998, 3298 patient specimens were compared by RETCIF and a modified shell vial method. Either or both methods isolated 779 viruses (24% positivity rate), whilst both methods detected 621. Of the 779 viruses, 87% (679) were isolated by the shell vial method in an average time of 4.9 days. For RETCIF the respective rate was 92.5% (721), in an average time of 3.0 days. The RETCIF method is a time-saving procedure, with higher isolation rates than the shell vial method. PMID- 11483220 TI - Predicting herd protection against foot-and-mouth disease by testing individual and bulk tank milk samples. AB - Four groups of cattle were tested for antibodies against foot-and-mouth disease (FMD) virus type O(1) over three 70 day vaccination cycles using the liquid-phase blocking-ELISA (LPBE). First lactation cows showed the lowest titres and group protection levels (GPLs) against FMD virus strains with 'r' values < or =0.5 while second lactation animals gave the highest results. When mean serum titres for each group and sampling date were plotted against GPL a strong correlation was found. Revaccination was indicated at a mean titre of approximately log10 2.88 (1:760; R=0.93; n=86) if the herd was threatened by field strains with an 'r' value of 0.25, or log10 2.62 (1:420; R=0.83; n=48) if this ratio was 0.5. Significant overall correlation (R=0.53; n=624) was obtained between serum titres and milk IgG(1) results derived from the modified specific isotype assay (SIA). Milk titres equivalent to 1:100, 1:200, 1:400 and 1:800 were 1:3.8, 1:6.3, 1:10.4 and 1:17.1, respectively, in first lactation cows. Bulk tank milk samples demonstrated a repeating pattern of results corresponding to the vaccination cycle with no titre lower than log10 1.05 (1:11). Colostrum from first lactation animals showed mean SIA results of log10 4.06 (1:11,480) and early milk titres only levelled off approximately 11 days post partum (dpp). PMID- 11483221 TI - Detection of bovine herpesvirus 4 glycoprotein B and thymidine kinase DNA by PCR assays in bovine milk. AB - A polymerase chain reaction (PCR) assay was developed to detect bovine herpesvirus 4 (BHV4) glycoprotein B (gB) DNA, and a nested-PCR assay was modified for the detection of BHV4 thymidine kinase (TK) DNA in bovine milk samples. To identify false-negative PCR results, internal control templates were constructed, added to milk samples, and co-amplified with viral DNA using the same primers for both templates. Specificity, sensitivity, and reproducibility of the two PCR assays were examined. In both PCR assays, all 31 BHV4 strains examined were scored positive, whereas 14 unrelated viruses scored negative. Sensitivity studies showed that two-ten copies of BHV4 DNA were detectable by the gB-PCR, while one-three copies could be detected by the TK-PCR. For the detection of BHV4 in milk samples, the gB-PCR amplification was found to be ten-times, and the TK PCR was found to be 55-times more sensitive than virus isolation. BHV4 DNA was detected by gB-PCR and TK-PCR in 93 and 95%, respectively, of 61 milk samples collected from cows infected intramammarily with BHV4, while only 61% were positive by virus isolation. Four out of 48 cows with clinical mastitis were positive for BHV4-gB and BHV4-TK DNA, whereas no BHV4 DNA was detected in milk from control cows. Considerable agreement was seen between the results of the two PCR assays, and both methods were considered as rapid and reliable tests for the screening of BHV4 DNA in bovine milk. The less laborious gB-PCR might be the recommended test of choice for screening large amounts of milk samples for the presence of BHV4. PMID- 11483222 TI - High-level expression of hepatitis C virus (HCV) structural proteins by a chimeric HCV/BVDV genome propagated as a BVDV pseudotype. AB - A chimeric cDNA genome was constructed in which the core, E1 and E2 genes of hepatitis C virus (HCV) replaced the core, E(rns), E1 and E2 genes of bovine viral diarrhea virus (BVDV). High levels of HCV structural proteins were expressed in a small number of human or bovine cells following transfection with chimeric RNA. However, in one cell line, bovine embryonic trachea cells [EBTr(A)], the number of cells expressing HCV proteins increased to greater than 70% following serial passage of culture medium. These cells were persistently infected with a non-cytopathogenic BVDV helper virus. In these cells, the chimeric genome was packaged into infectious particles that accumulated in the culture medium at a titer as high as 10(7)-10(9) genome equivalents per ml. The virus particles were pseudotypes, because they were neutralized by anti-BVDV but not by anti-HCV. PMID- 11483223 TI - Differentiating PVY(NTN) from PVY(N) by annealing to reference RNA transcripts. AB - A method for the differentiation of virus strains based on the shift in electrophoretic mobility of partially annealed RNA transcripts is described. Oppositely oriented RNA transcripts of the NTN- and N-strains of PVY, complementary at their 3'-end variable (strain-specific) region, were annealed to form a partial duplex which moved more slowly in gel than heterologous (NTN+N) unpaired transcripts. Thus, the two virus strains could be identified by annealing to a known reference transcript. The rate of duplex migration was correlated with transcript lengths and could be tightly controlled thereby. Thus, a higher degree of resolution was obtained than with transcript conformation polymorphism, which is empirical and unpredictable in nature. PMID- 11483224 TI - A recombinant particulate antigen of Japanese encephalitis virus produced in stably-transformed cells is an effective noninfectious antigen and subunit immunogen. AB - A COS-1 cell line, stably transformed by a plasmid encoding the premembrane and envelope glycoproteins of Japanese encephalitis virus, produced a noninfectious recombinant antigen expressed as extracellular particles. Extracellular particles purified by equilibrium density centrifugation in sucrose gradients followed by electron microscopy were characterized as spherical particles with an average diameter of approximately 30 nm and a buoyant density of 1.15 g/cc. Purified extracellular particles were shown by western blot to contain premembrane, membrane and envelope proteins. The gradient-purified particles exhibited hemagglutination activity at the same pH optimum (6.6) as Japanese encephalitis virus. Recombinant antigen from cell culture fluid was concentrated by precipitation with polyethylene glycol and evaluated for immunogenicity in 8-10 week-old ICR mice. Groups of five mice received only one immunization of recombinant antigen with or without Freund's incomplete adjuvant. Mice immunized with recombinant antigen plus Freund's incomplete adjuvant elicited the highest anti-viral titers as determined by both enzyme-linked immunosorbent assay (ELISA) and plaque-reduction neutralization tests. The polyethylene glycol-concentrated recombinant antigen was also evaluated for use in IgM antibody-capture ELISA and indirect IgG ELISA. The IgM-capture ELISA results using recombinant antigen correlated well with the results of a similar test using Japanese encephalitis virus-infected mouse brain antigen for the analysis of serum samples from patients with symptoms of acute encephalitis. Similar IgG titers were observed in an indirect ELISA comparing recombinant antigen and purified Japanese encephalitis virus as plate-bound antigens. Based on these studies, this entirely safe, easily produced antigen that expresses authentic Japanese encephalitis virus envelope glycoprotein would provide an excellent alternative to standard viral antigens used in various ELISA formats. PMID- 11483225 TI - Establishment of a MAGI-derived indicator cell line that detects the Nef enhancement of HIV-1 infectivity with high sensitivity. AB - The Nef protein of the simian and human immunodeficiency viruses (SIV and HIV) is regarded as one of the critical determinants of the pathogenicity of HIV-1 in vivo. The positive effect of Nef on viral replication is examined most easily in vitro by the use of indicator cells such as HeLa-CD4-LTR-beta-gal cells (MAGI) or MAGIC5 cells, which are MAGI-derived, CCR5-expressing cells. However, Nef increases the infectivity of many HIV-1 strains no more than 10-fold in these indicator cells. It was noted that MAGI cells expressing a lower level of CD4 enabled us to discriminate more clearly between wild-type and Nef-defective virions. A MAGIC5-derived cell line, MAGNEF, which stably expressed a low level of CD4, was established. The infectivity of the Nef-defective HIV-1 NL4-3 strain was consistently less than one-twentieth of that of the wild type in MAGNEF cells. By using MAGNEF cells, it was shown that Nef enhanced the infectivity of a subtype C HIV-1, Indie-C1 strain, although the effect of Nef on Indie-C1 was significantly less than that on the subtype B strains NL4-3 and SF2. These results validate the versatility of MAGNEF cells for use in the simple and sensitive assay for the level of Nef dependence of various HIV-1 isolates. PMID- 11483226 TI - Usefulness of blood and urine samples collected on filter paper in detecting cytomegalovirus by the polymerase chain reaction technique. AB - A rapid test for the diagnosis of congenital CMV infection is still needed. This study evaluated the usefulness of dried blood and urine samples collected on filter paper for detecting cytomegalovirus (CMV) by the polymerase chain reaction (PCR) assay compared with the use of liquid urine. Samples were obtained from 332 infants aged 1-7 days. Liquid urine samples were collected into bags, cultured in human fibroblasts, and processed using a multiplex PCR technique. Dried urine samples were obtained by placing a piece of filter paper in contact with the infant's genitals. The heels of neonates were punctured and capillary blood was blotted onto filter paper and dried. Dried blood and urine specimens were analyzed by multiplex PCR and nested-PCR assays. A diagnosis of congenital CMV infection was established by isolating the virus, and by detecting viral DNA in the liquid urine. Of the 332 liquid urine samples collected from 332 neonates, seven (2.1%) were positive for CMV and 325 were negative, by both cell culture and PCR assay. In dried samples, CMV DNA was detectable only with a nested PCR assay. Compared with known CMV infection status, 5/7 (71.4%) neonates were positive for congenital CMV infection using dried blood samples. All 325 uninfected neonates were negative. In the dried urine samples, 4/4 CMV-infected infants gave positive tests, and all 262 uninfected infants were negative. Although further improvements in sample collection and/or processing are still needed, PCR testing on dried urine or blood collected on filter paper is a promising approach in the diagnosis of neonatal CMV infection. PMID- 11483227 TI - Second-generation Hybrid capture test and Amplicor monitor test generate highly correlated hepatitis B virus DNA levels. AB - The performance of the Digene Hybrid Capture II HBV DNA Test HC II and the Roche Cobas Amplicor Monitor Test (Cobas-HBV) was evaluated on 252 serum samples. One hundred and seventy-three samples were HBV DNA positive and 75 HBV DNA negative by both assays. Four samples were HBV DNA positive by Cobas-HBV only. Linear regression analysis showed that the HBV DNA concentrations obtained from both assays were significantly related (n=173, r=0.976, P<0.0001). The results of the study show that Hybrid capture II and Cobas-HBV could be used equally in the management for patients with chronic HBV infection. PMID- 11483228 TI - The effect of extend bar containing uncooked cornstarch on night-time glycemic excursion in subjects with type 2 diabetes. AB - The objective of this study was to determine the effects of ingesting a snack bar containing uncooked cornstarch (Extend Bar, Clinical Products, Limited, Key Biscayne, FL) on nocturnal glycemic excursion in 28 adults (ages 22-78 years) with type 2 diabetes mellitus (mean HbAlc 8.21+/-1.28%). Thirteen subjects were treated with oral agents, eight with a combination of insulin and oral agents, and seven with insulin alone. Subjects ingested the study bar (Extend Bar, containing 30 g of total carbohydrate, including 5 g of uncooked cornstarch, 3 g protein, and 3 g fat) for three evenings followed by a placebo bar for 3 evenings (30 g of total carbohydrate, 3 g protein, and 3 g fat), or vice versa. Pre-snack before bedtime, midnight and before breakfast finger stick blood glucose levels were compared to determine the incidence of hypoglycemia (<60 mg/dl), hyperglycemia (>250 mg/dl), and to calculate any differences in the group's mean blood glucose levels when ingesting the study versus the placebo bar. There were no episodes of hypoglycemia or hyperglycemia. The mean blood glucose levels pre snack at bedtime were nearly identical (Extend Bar value 117.5+/-45.6 mg/dl; placebo bar value 117.3+/-40.0 mg/dl; P=0.977), and lower at midnight and before breakfast on the Extend Bar nights compared to the placebo bar nights (Extend Bar, midnight value 127.9+/-31.0 mg/dl; placebo bar midnight value 148.2+/-32.1 mg/dl; P=0.0001; Extend Bar breakfast value 114.2+/-15.8 mg/dl; placebo bar breakfast value 158.49+/-30.3 mg/dl; P<0.0001). These data suggest that ingesting Extend Bar containing uncooked cornstarch as the nighttime snack may be an effective strategy to lesson the frequency of nocturnal and morning hyperglycemia in subjects with type 2 diabetes. PMID- 11483229 TI - Repaglinide improves blood glucose control in sulphonylurea-naive type 2 diabetes. AB - The prandial glucose regulator repaglinide has a rapid onset of action, a short half-life and is metabolised mainly by the liver. Here we report the findings of a 10-week, double-blind, parallel, placebo controlled, randomised trial with repaglinide in 25 diet-treated, sulphonylurea-naive patients with Type 2 diabetes. Repaglinide was titrated, based on capillary blood glucose, from 0.5 mg to a maximum of 4 mg, preprandially with breakfast and dinner. After 10 weeks, repaglinide was associated with a decrease in HbA(1c) of 2.3%Hb relative to the placebo group (P=0.018). This reflected a 30% decrease within the repaglinide group from a mean HbA(1c) of 7.0 to 4.9%Hb (P<0.002). Repaglinide was also associated with a decrease in fructosamine, by 0.88 mmol/l, relative to placebo (P<0.001), with a 20% decrease (from 3.80 to 3.04 mmol/l) within the repaglinide group (P<0.001). Fasting and postprandial blood glucose concentrations decreased in association with repaglinide by 3.6 and 6.4 mmol/l, respectively, relative to placebo (P<0.001 in each case). Within the repaglinide group fasting and postprandial blood glucose decreased by 3.9 and 6.2 mmol/l, respectively (P<0.001 in each case). The number of patients reporting hypoglycaemia in the repaglinide group was similar to placebo (15 vs. 20, respectively; NS). Test meal assessments confirmed that repaglinide effectively controls glucose levels by stimulating mealtime insulin secretion. Fasting serum insulin concentration was not raised compared to baseline or placebo during repaglinide therapy, albeit that fasting glucose levels were decreased by repaglinide. Twice-daily meal-related insulin secretagogue therapy with repaglinide, a new short and rapid-acting prandial glucose regulator, is capable of improving all measures of glycaemic control without increased hypoglycaemia or fasting hyperinsulinaemia. PMID- 11483230 TI - Nitric oxide system is involved in glomerular hyperfiltration in Japanese normo- and micro-albuminuric patients with type 2 diabetes. AB - Glomerular hyperfiltration plays a pathogenic role in the early stages of diabetic nephropathy. Experimental studies in laboratory animals suggest that nitric oxide (NO) might be involved in the pathogenesis of glomerular hyperfiltration. We performed a cross-sectional study to determine the relationship between diabetic glomerular hyperfiltration and the NO system. Normoalbuminuric (n=41), microalbuminuric (n=25), and macroalbuminuric (n=16) patients with type 2 diabetes were recruited in this study and compared with age matched 84 non-diabetic control subjects. Creatinine clearance and urinary NO(2)( )/NO(3)(-) excretion (urinary NOx) were measured, and the expression of endothelial cell nitric oxide synthase (ecNOS) was evaluated in human renal tissues. Glomerular hyperfiltration was present in 19 (37.5%) and nine (36.6%) of normoalbuminuric and microalbuminuric type 2 diabetic patients, respectively. The urinary NOx was significantly higher in normoalbuminuric patients compared with normal subjects. Creatinine clearance correlated significantly with urinary NOx in normoalbuminuric and microalbuminuric patients. Immunohistochemical staining intensities for ecNOS were significantly increased in glomerular endothelial cells of microalbuminuric type 2 diabetic patients as compared with the control subjects. These results suggest that NO may contribute to the pathogenesis of glomerular hyperfiltration in Japanese type 2 diabetic patients. PMID- 11483231 TI - Serum leptin level as an indicator to predict the clinical efficacy of troglitazone in patients with type 2 diabetes mellitus. AB - Troglitazone is effective in approximately 50% in patients with type 2 diabetes (NIDDM). In this study, we investigated the relations between serum leptin levels and clinical efficacy of troglitazone. Forty-five type 2 diabetic patients (23 men and 22 women) from our outpatient clinic were treated with troglitazone 400 mg daily for 12 weeks. Fasting plasma glucose (FPG), HbA1c, body weight, serum insulin and leptin concentrations were measured before and after troglitazone treatment. After 12 weeks of troglitazone treatment, FPG (before versus after, 179+/-33 vs. 138+/-26 mg/dl, mean+/-SD), HbA1c (7.8+/-1.3 vs. 6.9+/-1.0%), IRI (8.3+/-4.3 vs. 6.3+/-3.4 microU/ml) and HOMA-R index (homeostasis model assessment of insulin resistance) (3.8+/-2.4 vs. 2.2+/-1.3) decreased significantly, while body mass index (BMI) slightly increased (26.3+/-3.5 vs. 26.6+/- 3.6 kg/m(2)), and serum leptin remained unchanged (8.5+/-7.2 vs. 9.1+/ 8.7 ng/ml). Reduction in FPG (DeltaFPG) after troglitazone treatment were correlated with reduction in HOMA-R (DeltaHOMA-R) (r=0.721, P<0.0001). DeltaFPG was correlated with serum leptin (r=0.441, P<0.01), HOMA-R (r=0.460, P<0.01) and FPG (r=-0.781, P<0.0001) at baseline, but not with BMI and serum IRI at baseline. Furthermore, serum leptin at baseline was significantly correlated with DeltaHOMA R (r=0.634, P<0.01). Leptin concentration before treatment therefore, can be used as an predictor for clinical efficacy of troglitazone in patients with type 2 diabetes. PMID- 11483232 TI - Dementia and its associations in type 2 diabetes mellitus: the Fremantle Diabetes Study. AB - BACKGROUND: recent studies indicate that diabetes is an important risk factor for dementia in older patients, but the cause remains unknown. OBJECTIVES: to determine whether vascular or diabetes-related risk factors predict the development of dementia in older subjects with diabetes. PATIENTS: 63 patients with type 2 diabetes of mean age 75.3 years. METHODS: Subjects were screened for cognitive impairment using the Mini-Mental State Examination (MMSE) and informants who knew the subjects answered the Informant Questionnaire for Cognitive Decline in the Elderly (IQCODE). Probable dementia was diagnosed using highly specific criteria based on the combination of both tests. Potential risk factors for dementia obtained at the time of the cognitive test and annually from a median of 3.2 years previously were examined using univariate methods and simple general linear modelling. RESULTS: since there was a significant association between MMSE and English speaking ability, ten non-Anglo-Celt patients were excluded from the analysis. Probable dementia was diagnosed in six of the remaining 53 subjects (11%). Dementia was significantly and independently associated with higher diastolic and mean arterial blood pressure measurements over the 3 years before assessment. Blood pressure declined over this time in the patients with probable dementia and was similar to that in non-demented subjects at the time of cognitive assessment. CONCLUSIONS: these preliminary data suggest that the high rate of dementia found in older people with diabetes may be explained by the high rate of hypertension in this population. PMID- 11483233 TI - Screening for people with diabetes mellitus for poor blood glucose control in an ophthalmological laser clinic. AB - PURPOSE: The study was performed to test the effect of a structured intervention in diabetic patients with poor glycaemic control in an ophthalmological department. PATIENTS AND METHODS: All the patients attending the ophthalmological out-patient department with the need for laser therapy due to diabetic retinopathy were investigated from January to March 1998 (Type 1: n=20, Type 2: n=144). If an HbA(1c)-level higher than 9.0% was found the patient was informed within 1 week and a standardised letter was sent to the primary care physician and the local ophthalmologist. Over the first 3 months of 1999 the effect was evaluated. RESULTS: HbA(1c) values higher than 9.0% were found in eight/20 of the patients (40%) with Type 1 diabetes and in 61/144 of the patients (54%) with Type 2 diabetes. In 55% this new information in the context of the need for laser therapy resulted in the acceptance of a structured intervention by the patient. This led to an improvement of the HbA(1c) in the people with Type 1 diabetes. CONCLUSION: Patients with poor blood glucose control can be identified in an ophthalmological department. The need for laser therapy can be used to motivate the patients for a significant improvement of the quality of blood glucose control. PMID- 11483234 TI - Prevalence of urinary tract infection and renal scars in patients with diabetes mellitus. AB - In a case control study, we assessed the prevalence of bacterial urinary tract infections (UTI) and renal scarring in 155 consecutive type 1 (n=102) and type 2 (n=53) diabetic individuals and 128 healthy controls. Subjects who received antibiotics during the past 6 months, pregnant women and those with overt renal failure were excluded. In all subjects, urine culture and 99m Technetium (Tc) dimercapto-succinic acid renal scan was performed. UTI was diagnosed if two consecutive urine cultures grew the same organism with at least 10(5) colony forming unit (cfu)/ml in asymptomatic and at least 10(4) cfu/ml in symptomatic subjects, respectively. Renal scan was considered abnormal if focal or multiple tracer uptake defects and/or break in cortical outline were observed. The prevalence of UTI in diabetes mellitus was higher, when compared to that in controls (9% vs. 0.78%, P=0.005). Escherichia coli was the most commonly grown organism (64.3%), followed by Staphyloccocus aureus (21.4%) and Klebsiella pneumoniae (14.3%). Prevalence of renal scarring was higher in patients with diabetes (28/155, 18.0%), when compared to that of controls (7/128, 5.4%, P=0.002). Fifty percent of patients with diabetes and UTI had renal scarring. The prevalence in diabetics with no UTI was also higher, when compared to controls (14.8 vs. 5.5%, P<0.01). The prevalence of UTI as well as renal scarring was significantly higher in females, when compared to male diabetics. No significant difference in vascular events, hypertension, proteinuria, renal function tests and HbA1 was observed in patients with and without renal scar. Thus, patients with diabetes mellitus have 10- and 3-folds increased risk of UTI and renal scarring, respectively. The results could help prioritize protocols for management of UTI among patients with diabetes mellitus. PMID- 11483235 TI - The adequacy of diabetic care for children in a developing country. AB - A cross-sectional study was conducted over 4 months evaluating the quality of care provided to diabetic children in public children's hospitals in Alexandria, Egypt. RESULTS: Adult diabetologists were the main healthcare providers (HCP) (60.4%) in the School Health Insurance Hospital followed by paediatric diabetologists in the University Hospital. Insured children had a significantly higher frequency of physical examination, investigations and diabetes education compared to uninsured children. One-quarter of insured and 22% of uninsured children were performing self monitoring of blood glucose, while 45.2% of insured children were checking glucosuria at home compared to 34.0% of uninsured children. Premixed suspensions of biosynthetic human insulin, administered mainly via a syringe, was the most commonly prescribed insulin type with little possibility for personal initiative. Acute diabetic complications were also higher in uninsured compared to insured children. The frequency of these life threatening acute diabetic complications in the school health insurance system is estimated to be approximately 12.7 severe hypoglycaemic and 57.2 hyperglycaemic/ketoacidotic episodes per 1000 diabetic children per year. Recurrence of diabetic emergencies was significantly higher among children of parents with lower educational levels and children living in semiurban and rural residence. Children with recurrent diabetic emergencies had lower educational achievement, and more grade repeating and school absence during the year. CONCLUSION: The results of this study appear to reflect marked deficiencies in the provision of information to children with diabetes and their parents in a developing country. A need for public-education strategies, consensus about treatment recommendations, use of more flexible insulin regimens, and devices for home monitoring is identified. PMID- 11483236 TI - Differential expression patterns of three glutamate transporters (GLAST, GLT1 and EAAC1) in the rat main olfactory bulb. AB - Glutamate is the main neurotransmitter in the olfactory bulb. Therefore, glutamate transporters, which regulate the concentration of extracellular glutamate, might play pivotal roles in odor processing. In this study, we examined expressions of three glutamate transporters (GLAST, GLT1 and EAAC1) in the olfactory bulb using in situ hybridization and immunohistochemistry. EAAC1 mRNA was expressed in neurons, such as periglomerular cells, tufted cells, mitral cells and granule cells as shown before in other brain areas. In contrast, GLAST and GLT1 were found in glial cells throughout the olfactory bulb, with intenser expressions in the glomerular layer, external plexiform layer and internal plexiform layer where glutamatergic synapses are concentrated. In addition, using double staining immunohistochemistry we clearly showed that GLAST and GLT1 were expressed in astrocytes. Furthermore, we found that GLAST was also intensely expressed in the subependymal layer where precursor cells exist. These results suggest each glutamate transporter plays its unique role not only in glutamatergic neurotransmission but also in cell differentiation and migration in the olfactory bulb. PMID- 11483237 TI - Suppression of Ca2+ influx through L-type voltage-dependent calcium channels by hydroxyl radical in mouse cerebral cortical neurons. AB - In the present study, we investigated the effect of hydroxyl radical (.OH) produced by the Fenton reaction with FeSO(4) to H(2)O(2) on Ca2+ influx by measuring [(45)Ca2+] influx into mouse cerebral cortical neurons in primary culture.OH formed from 3 microM FeSO(4) and 0.01 microM H(2)O(2) significantly reduced 30 mM KCl-induced [(45)Ca2+] influx and this reduction was abolished by .OH scavengers such as N,N'-dimethylthiourea and mannitol. Nifedipine (1 microM), an inhibitor for L-type voltage-dependent Ca2+ channels (VDCCs) showed no additive effect on the reduction of the 30 mM KCl-induced [(45)Ca2+] influx, while the inhibitors for P/Q- and N-type VDCCs showed further suppression of the KCl-induced [(45)Ca2+] influx even in the presence of .OH. Bay k 8644, an activator of L-type VDCCs, dose-dependently stimulated [(45)Ca2+] influx, and this stimulation disappeared in the presence of nifedipine. Similarly, .OH also suppressed significantly [(45)Ca2+] influx induced by Bay k 8644. These inhibitory actions of .OH on the KCl- and Bay k 8644-induced [(45)Ca2+] influx were completely abolished by .OH scavengers. These results indicate that .OH has the activity to suppress Ca2+ influx through L-type VDCCs. PMID- 11483238 TI - Analysis of human Per4. AB - The molecular mechanism of the circadian pacemaker depends on the oscillatory expression of clock gene constituents. The Drosophila period gene is central to the clock mechanism in these animals. Three homologs of this gene identified in mice (mPer1-3) and humans (hPer1-3) display rhythmic expression and are important for normal clock function. Recently, analysis of the draft sequence of the human genome has revealed the presence of a fourth Per gene family member. Surprisingly, the deduced hPer4 cDNA has no open reading frame encoding a full length PER-like protein. This sequence is characterized by numerous deletions, insertions, frame shifts and base pair changes, and its genomic structure is devoid of introns. The presence of an MER-2 mobile element fossil within the Per4 locus predicted that this gene would also be present in non-human primates. Rhesus monkey Per4 displays similar sequence anomalies and is 92.8% identical to hPer4. Sequence comparisons indicate that Per4 originated from a Per3 predecessor and that it is relatively new to the Period gene family. We conclude that hPer4 and RmPer4 are pseudogenes and descended from the retrotransposition of an ancestral Per3 gene. PMID- 11483240 TI - Central and peripheral dysregulation of melanin-concentrating hormone in obese Zucker rats. AB - Melanin concentrating hormone (MCH) is a peptide synthesized in the lateral hypothalamus which stimulates food ingestion and leptin secretion in rodents. In this experiment, we measured the expressions of MCH as well as of its receptor (SLC-1) in the hypothalamus of obese hyperphagic and lean Zucker rats by quantitative real time RT-PCR. MCH mRNA expression in the obese rats was significantly increased by a factor of five (P<0.01) whereas expression of SLC-1 was decreased by more than 50% (P<0.05). Circulating levels of leptin and MCH were increased in the plasma of obese Zucker rats when compared to lean rats (38 fold and 1.7-fold, respectively, P<0.001 and P<0.01). However, individual MCH levels were not directly correlated to leptin levels in the lean (functional leptin receptor) or in the obese (non-functional leptin receptor) Zucker rats. These results indicate that the absence of leptin signaling in rats is associated with an increased hypothalamic expression and circulating release of MCH, contributing to their obesity syndrome. PMID- 11483239 TI - Neuronal expression of mint1 and mint2, novel multimodular proteins, in adult murine brain. AB - Mints are multimodular adapter proteins in functioning membrane transport and organization. Mint1 and mint2 are neuron-specific. We localized these isoforms in mouse brain. By in situ hybridization, mRNA encoding mint1 or mint2 was expressed in neurons throughout the brain. Mint1 mRNA expression was greatest in the limbic system including cingulate cortex, hippocampus, anterior thalamic nuclei, medial habenular nucleus, and mammillary body. Mint2 mRNA was rich in cerebral cortex, entorhinal cortex, and hippocampus, but less prominent in other limbic structures. Mint1 mRNA and mint2 mRNA were distributed among hippocampal pyramidal neurons, while mint2 mRNA was especially abundant in CA3. Mint1, but not Mint2 mRNA was abundant in the substantia nigra pars compacta. Immunohistochemistry visualized mint proteins in axon terminals and neuronal somata, generally following mRNA distribution. In the hippocampus, mint1 was rich in the entorhinal projections and mossy fibers of the dentate gyrus, while mint2 was rich in commisural fibers from the contralateral hippocampus and in CA1. Mint1 intensely stained catecholamine-containing neurons such as the substantia nigra pars compacta, ventral tegmental area, and locus ceruleus. Mint2 protein was ubiquitous in these regions. Mint1 and mint2 distribution also differed elsewhere in the brainstem and in the cerebellum. Central nervous system neurons, then, predominantly express either mint1 or mint2. Mints may be involved in synaptic vesicle transport toward the active zone, also participating in transport of certain membrane proteins toward the postsynaptic density. Mint1 and mint2 may divide roles either regionally or depending on neuronal functional characteristics. PMID- 11483241 TI - A null mutation in guanylate cyclase-1 alters the temporal dynamics and light entrainment properties of the iodopsin rhythm in cone photoreceptor cells. AB - Guanylate cyclase-1 (GC1) plays a critical role in visual phototransduction and its absence severely compromises the ability of the photoreceptor cells to transduce light for vision. In this study we sought to determine if the absence of GC1 has any effect on light entrainment of the circadian oscillators located in these cells. We compared the rhythmic changes in transcript levels of iodopsin, a photoreceptor-specific gene whose expression is regulated by circadian oscillators, in retinas of normal chickens and GUCY1*B (*B) chickens that carry a null mutation in GC1. Our results show that iodopsin rhythms are present in *B retinas and that they can be entrained to light; however, the rise and fall of iodopsin transcript levels in *B retina under cyclic light conditions is significantly more rapid than that observed in normal retina, and under constant dark conditions, the phase of the iodopsin rhythm in *B retina is advanced by 6 h relative to that observed in normal retina. In addition, the rate of entrainment of the iodopsin rhythm in *B retina to a reversal of the light cycle is significantly slower than normal. The results of our study show that a functioning visual phototransduction cascade is not essential for light entrainment of the oscillators that drive the iodopsin rhythm in photoreceptor cells. We propose that the abnormal synthesis of cGMP in *B photoreceptors underlies the irregular iodopsin rhythms observed in post-hatch *B retina. PMID- 11483242 TI - Expression of alpha-synuclein in the human brain: relation to Lewy body disease. AB - Alpha-synuclein is mutated in some hereditary cases of Parkinson's disease and the protein precipitates in Lewy bodies, the pathological hallmark of both Parkinson's disease and Lewy body disease. Transgenic mice overexpressing human wild-type alpha-synuclein develop alpha-synuclein-immunoreactive inclusions in brain regions typically affected with Lewy body disease. We used in situ hybridization to characterize alpha-synuclein expression and examine mRNA levels in patients affected with Lewy body disease and controls. Substantia nigra was avoided because of the extensive neuronal loss and cingulate gyrus was chosen as it is one of the diagnostic regions in Lewy body disease where Lewy bodies most frequently are demonstrated. beta-tubulin was used to control for neuronal degeneration. The alpha-synuclein probe showed intense labeling of pyramidal cells in lamina III and V in both patients and controls. We found no difference in alpha-synuclein mRNA levels and beta-tubulin mRNA was not significantly altered (P=0.06) in patient brains. There was no difference in the ratio of alpha synuclein and beta-tubulin mRNA levels between patients and controls. Further, we found no relationship between alpha-synuclein mRNA levels and Lewy bodies. Great variability in alpha-synuclein mRNA levels among patients indicates that Lewy body disease may be a heterogeneous disorder with regard to alpha-synuclein involvement. PMID- 11483243 TI - Serotonin 2A receptor regulation of striatal neuropeptide gene expression is selective for tachykinin, but not enkephalin neurons following dopamine depletion. AB - Serotonin (5-HT) 2A receptor-mediated regulation of striatal preprotachykinin (PPT) and preproenkephalin (PPE) mRNAs was studied in adult rodents that had been subjected to near-total dopamine (DA) depletion as neonates. Two months following bilateral 6-hydroxydopamine (6-OHDA) lesion, PPT mRNA levels decreased 59-73% across dorsal subregions of the rostral and caudal striatum while PPE transcripts increased 61-94%. Four hours after a single injection of the serotonin 2A/2C receptor agonist, (+/-)-1-(2,5-Dimethoxy-4-iodophenyl)-2-aminopropane (DOI; 1 mg/kg), PPT mRNA expression was significantly increased in DA-depleted rats across all dorsal subregions of the rostral and caudal striatum as compared to 6 OHDA-treated animals alone. In the intact rat, DOI did not influence PPT mRNA levels in the rostral striatum, but did raise expression in the caudal striatum where 5-HT2A receptors are prominent. DOI did not regulate PPE mRNA levels in any striatal sub-region of the intact or DA-depleted rat. Prior administration of the 5-HT2A/2C receptor antagonist, ritanserin (1 mg/kg) or the 5-HT2A receptor antagonist, ketanserin (1 mg/kg) completely blocked the DOI-induced increases in striatal PPT mRNA in both lesioned and intact animals. The ability of ketanserin to produce identical results as ritanserin suggests that 5-HT2A receptor-mediated regulation is selectively strengthened within tachykinin neurons of the rostral striatum which are suppressed by DA depletion. The selectivity suggests that 5 HT2A receptor upregulation following DA depletion is capable of regulating tachykinin biosynthesis without influencing enkephalin expression in striatal output neurons. PMID- 11483244 TI - Altered emotional behavior in PACAP-type-I-receptor-deficient mice. AB - PAC1 (pituitary adenylate cyclase activating polypeptide type I receptor) is a G protein-coupled receptor that binds the strongly conserved neuropeptide PACAP (pituitary adenylate cyclase activating polypeptide) with a thousandfold higher affinity than the related peptide VIP (vasoactive intestinal peptide). PAC1 shows strong expression in brain areas which have been implicated in the emotional control of behavior, such as the amygdala, the hypothalamus, the locus coeruleus and the periaqueductal gray. To assess whether PAC1-mediated signaling has an impact on emotional behavior, we analysed two different mutant mouse lines with an ubiquitous or a forebrain-specific inactivation of PAC1 in several testing paradigms modelling general locomotor activity and anxiety-related behavior. We clearly demonstrate that mice with a ubiquitous but not with a forebrain-specific deletion of PAC1 exhibit elevated locomotor activity and strongly reduced anxiety like behavior. We could not observe any gross alteration in circadian rhythmicity nor any enhanced sensitivity towards ethanol in the mutant mice. We previously demonstrated that PAC1 plays a crucial role in contextual fear conditioning. Therefore the finding that PAC1-deficient mice exhibit reduced anxiety is quite exciting, since the receptor and hence its ligand PACAP seem to be important for both, innate and learned fear. PMID- 11483245 TI - Functional characterization of the human Huntington's disease gene promoter. AB - The genetic basis of neurodegeneration in Huntington's disease (HD) has been identified as a (CAG)(>37) repeat expansion in a gene of unknown function. Interestingly, patients with the same expanded (CAG)(n) repeat length may have markedly different ages at onset. Based on experiences in animal models the level of expression might be one of the modifying factors. To gain insight into the regulation of the human HD gene we functionally analyzed 2266 bp of the HD gene promoter region. This region lacks a TATA and a CAAT box, is GCrich, and it has several consensus sequences for SP1, AP-2 and AP-4 binding sites. The stretch between nucleotides -49 and -198 relative to the first ATG is highly conserved between human and rodents and it harbors several potential binding sites for transcription factors. We analyzed deletion mutants fused with the chloramphenicol acetyltransferase (CAT) reporter gene in transfected, huntingtin expressing neuronal (NS20Y) and non-neuronal (CHO) cell lines. Partial deletion of the evolutionarily conserved part of the promoter significantly reduces the activity in both neuronal and non-neuronal cells indicating that the core promoter activity is located between nucleotides -221 and 4, relative to the +1 translation start site. Binding affinities of DNA-protein interactions were defined by electrophoretic mobility shift assays and the protected nucleotide positions were determined by DNase I footprinting. PMID- 11483246 TI - The cyclic AMP response element in the Bcl-2 promoter confers inducibility by hypoxia in neuronal cells. AB - In neuronal cells, expression of the anti-apoptotic Bcl-2 gene is induced by hypoxia and produces a protective effect. We show here that this effect is dependent upon the cyclic AMP response element (CRE) in the Bcl-2 promoter since mutation of this element abolishes the response and the isolated CRE can confer the response on a heterologous promoter. Interestingly however, the CRE in the Bcl-2 promoter does not render the promoter responsive to cyclic AMP and is not essential for its response to nerve growth factor. Despite the lack of cyclic AMP responsiveness, activation of the Bcl-2 promoter via the CRE in response to hypoxia requires the CREB transcription factor and is associated with the enhanced phosphorylation of CREB on serine 133 and enhanced transcriptional activation by the CREB-binding protein, CBP, in response to hypoxia. This finding establishes the importance of the CRE in the induction of Bcl-2 gene expression by hypoxia, allowing the Bcl-2 protein to protect neuronal cells against this damaging stimulus. PMID- 11483247 TI - Sequential changes in AMPA and NMDA protein levels during Fe(3+)-induced epileptogenesis. AB - Seizure susceptibility is related to enhanced glutamatergic excitatory synaptic transmission with alterations in the expressions of ionotropic glutamate receptors. We wondered if levels of AMPA and NMDA receptor subunits changed following epileptogenesis induced by amygdalar FeCl(3) injection. We used Western blots to measure levels of subunits in the ipsilateral and contralateral hippocampus at various times after FeCl(3) injection into the amygdaloid body. With acute seizures, at +5 days after the injection, levels of GluR1, NMDAR1, and NMDAR2 were markedly increased in both hippocampi, with quantities at least 2-4 times baseline. By +15 and +30 days after injection, when chronic spontaneous seizures were occurring, the levels of GluR2 were increased, while GluR1 and NMDAR1&2A/B were decreased. Increased NMDAR1&2A/B levels at +5 days are consistent with the occurrence of upregulation of NMDA receptor production in the early stages of epileptogenesis. Since GluR2 suppresses glutamate receptor mediated Ca(2+)-influx, increased expression of GluR2 with development of chronic, recurrent seizures may be a compensatory effect during epileptogenesis from neural responses to propagated seizures. PMID- 11483248 TI - Persistent corticotropin-releasing factor(1) receptor desensitization and downregulation in the human neuroblastoma cell line IMR-32. AB - Brain corticotropin-releasing factor (CRF) systems integrate various responses to stress. Pathological responses to stress may result from errors in CRF receptor regulation in response to changes in synaptic CRF levels. To establish an in vitro model to study brain CRF receptors, we characterized the CRF-induced modulation of CRF(1) receptors in the human neuroblastoma cell line, IMR-32. Treatment with CRF decreased CRF(1) receptor binding and desensitized CRF-induced increases in cAMP. The decrease in binding had an EC(50) of approximately 10 nM, was maximal by 30 min, and was blocked by the CRF receptor antagonist [D-Phe(12), Nle(21,38), C(alpha)-MeLeu(37)]CRF(12-41). The desensitization was homologous as vasoactive intestinal polypeptide-induced increases in cAMP were unchanged, and elevation of cAMP did not alter CRF(1) receptor binding. Treatment with CRF for up to 24 h did not alter CRF(1) receptor mRNA levels, suggesting that a posttranscriptional mechanism maintains the decrease in receptor binding. Interestingly, recovery of CRF receptor binding and CRF-stimulated cAMP production was only partial following exposure to 100 nM CRF. In contrast, receptor binding recovered to control levels following exposure to 10 nM CRF. These data suggest that exposure to high doses of CRF result in permanent changes characterized by only partial recovery. Identifying the mechanisms underlying this partial recovery may provide insights into mechanisms underlying the acute and chronic effects of stress on CRF receptor regulation. PMID- 11483249 TI - Effects of experimental diabetes on axonal and Schwann cell changes in sciatic nerve isografts. AB - A reduced ability to regenerate peripheral axons may be partly responsible for diabetic neuropathy. The source of the impairment has not been narrowed down to axonal or Schwann cell failure. We used nerve grafts from control or diabetic donor rats transplanted into control or diabetic hosts to pursue this differential diagnosis. An isograft between the left sciatic nerves of inbred Lewis rats was performed 8 weeks after STZ treatment and on age-matched controls. The nerve exchanges were control-control, control-diabetic, diabetic-control and diabetic-diabetic. At postsurgical day 14, nerves were excised and analysed for levels of axonal markers, total and phosphorylated neurofilament, and Schwann cell receptors, ErbB2 and p75(NTR), using immunohistochemistry and Western blotting. The aim was to measure ingress of axonal markers into the graft and judge the appropriateness of Schwann cell phenotype changes. Transfer of nerve from diabetic to control rats resulted in a doubling in neurofilament, both phosphorylated and nonphosphorylated (both P<0.05). ErbB2 was decreased in grafts from diabetic rats (53% of control, P<0.05) and p75(NTR) levels were increased in both types of graft in diabetic rats (to 300-400% of controls, P<0.05). Schwann cells in diabetic nerve grafts showed receptor levels more similar to controls when placed into a normal environment and the converse also appeared to hold. TUNEL staining revealed increased apoptosis in diabetic nerve distal to the graft. The data show that alterations in Schwann cell phenotype in diabetes are reversed by transfer to control rats and develop in normal nerve after transfer to a diabetic host. PMID- 11483250 TI - Increase in expression of the GABA(A) receptor alpha(4) subunit gene induced by withdrawal of, but not by long-term treatment with, benzodiazepine full or partial agonists. AB - The effects of long-term exposure to, and subsequent withdrawal of, diazepam or imidazenil (full and partial agonists of the benzodiazepine receptor, respectively) on the abundance of GABA(A) receptor subunit mRNAs and peptides were investigated in rat cerebellar granule cells in culture. Exposure of cells to 10 microM diazepam for 5 days significantly reduced the amounts of alpha(1) and gamma(2) subunit mRNAs, and had no effect on the amount of alpha(4) mRNA. These effects were accompanied by a decrease in the levels of alpha(1) and gamma(2) protein and by a reduction in the efficacy of diazepam with regard to potentiation of GABA-evoked Cl- current. Similar long-term treatment with 10 microM imidazenil significantly reduced the abundance of only the gamma(2)S subunit mRNA and had no effect on GABA(A) receptor function. Withdrawal of diazepam or imidazenil induced a marked increase in the amount of alpha(4) mRNA; withdrawal of imidazenil also reduced the amounts of alpha(1) and gamma(2) mRNAs. In addition, withdrawal of diazepam or imidazenil was associated with a reduced ability of diazepam to potentiate GABA action. These data give new insights into the different molecular events related to GABA(A) receptor gene expression and function produced by chronic treatment and withdrawal of benzodiazepines with full or partial agonist properties. PMID- 11483251 TI - Ribozyme-mediated reduction of the GABA(A) receptor alpha1 subunit. AB - As an approach to understanding the role of the alpha1 subunit of the GABA(A) receptor, ribozymes were designed to reduce expression of this subunit protein by hydrolysis of alpha1 subunit message and antisense inactivation. The ribozyme cleavage sites were selected through homology comparison of all known murine GABA(A) receptor subunits at the amino acid and nucleotide sequence level. Two ribozymes were designed and synthesized: one against the extracellular domain and the other against the cytoplasmic domain. These ribozymes were cloned in a mammalian expression plasmid, pZeoSV2 (+). Cleavage of both extracellular and cytoplasmic domain transcripts by the respective ribozymes was observed when each ribozyme was tested against in vitro transcribed mRNA. The stable cell line, 122, expressing recombinant human GABA(A) alpha1, beta2 and gamma2S subunits of receptor was stably transfected with the cytoplasmic domain ribozyme (cy) alone and with both the cytoplasmic (cy) and extracellular domain (ex) ribozyme expression plasmids. Northern analysis showed a 55-60% reduction of alpha1 mRNA in clones of cells transfected with either the single ribozyme (Cy) or with both ribozymes (EC). The alpha1 protein level was reduced 75% in a stable Cy clone and more than 90% in a stable EC clone when compared with alpha1 expression in 122 cells and the vector transfected (Zeo) cells. Electrophysiological analysis revealed that the GABA(A) receptor properties were very similar in 122 cells and in stable clones in which the subunit protein expression had been greatly reduced. No significant difference was detected in the potentiation of the receptor response by either bretazenil or zolpidem. These data demonstrate the efficacy of the ribozyme approach in dramatically reducing GABA(A) subunit protein levels in transfected cells and identify those elements that will be important to the application of similar ribozymes to knock-down transmitter receptor subunit proteins under inducible promoters in transgenic mice. PMID- 11483252 TI - Transient changes of brain-derived neurotrophic factor (BDNF) mRNA expression in hippocampus during moderate ischemia induced by chronic bilateral common carotid artery occlusions in the rat. AB - Chronic bilateral common carotid artery occlusion (BCCAO) induces moderate ischemia (oligemia) in the rat forebrain in the absence of overt neuronal damage. In situ hybridization for brain-derived neurotrophic factor (BDNF) mRNA was used to search for a molecular response to moderate ischemia. BDNF mRNA was significantly increased in the hippocampal granule cells at 6 h of occlusion (ANOVA, Tukey test P<0.05). At 1, 7 and 14 days BDNF mRNA levels returned to control levels. The frequency of BDNF gene expression at 6 h was 83%, which was significantly higher than the 7% incidence of histological injury in the hippocampus (Fisher's exact test, P<0.002). Cerebral blood flow was reduced to 75% of control levels in the hippocampus after 1 week of BCCAO when measured with the autoradiographic method. Measurements of tissue flow with a microprobe for laser Doppler flow excluded decreases into the ischemic range during the period when elevated gene expression was observed. Prolonged moderate ischemia (oligemia) is a sufficient stimulus for BDNF gene expression in the hippocampus. These molecular studies provide direct evidence for an involvement of the hippocampus in the BCCAO model. PMID- 11483253 TI - Differential usage of multiple brain-derived neurotrophic factor promoter in rat dorsal root ganglia following peripheral nerve injuries and inflammation. AB - The brain-derived neurotrophic factor (BDNF) may act as either an autocrine or paracrine survival factor for the dorsal root ganglion (DRG) neurons and may also serve as a neurotransmitter or neuromodulator within the dorsal horn of the spinal cord. The rat BDNF gene consists of four short 5' exons linked to separate promoters and one 3' exon encoding the mature BDNF protein. An exon-specific reverse transcription-polymerase chain reaction analysis was used to study the differential utilization of multiple promoters in the DRG following unilateral sciatic axotomy, dorsal rhizotomy and peripheral inflammation. The exon I transcript showed the highest induction rate with the ipsilateral expression elevated 4.3-5.8 times that of contralateral expression. Both exon II and III mRNAs showed a smaller increase 1 day after the three kinds of stimuli. In addition, exon IV mRNA transcription increased slightly only after rhizotomy, but not after axotomy and peripheral inflammation after 1 day. Furthermore, the elevated exon I mRNA levels 1 day after rhizotomy were sustained for up to 7 days. In contrast, those of the exon I mRNA after axotomy had declined 2.8 times the control level after 7 days. These findings suggest that the promoter linked to exon I may provide a major regulatory point of BDNF mRNA expression by peripheral nerve injuries and inflammation. In addition, both exon I and IV mRNA expression may show different temporal activation patterns according to the types of injury. PMID- 11483254 TI - Retinoic acid receptors are expressed in human primary astrocytes and their agonists inhibit parathyroid hormone-related peptide expression and secretion. AB - We isolated, by RT-PCR, partial cDNAs encoding retinoic acid receptor (RAR) isoforms, alpha, beta and gamma, in human primary astrocytes and confirmed their expression by western analysis. A naturally occurring RAR agonist, all-trans retinoic acid and a synthetic RAR agonist, [(E)-4-[2-(5,6,7,8-tetrahydro-5,5,8,8 tetramethyl-2-naphthylenyl)-1-propenyl]benzoic acid], each inhibited PTHrP secretion with EC(50) values of approximately 25 and 250 nM, respectively. The two may act at a transcriptional level as assessed by Northern analysis. Thus retinoids, most likely acting via RARs, inhibit parathyroid hormone-related protein expression and secretion in human primary astrocytes, with a synthetic RAR-specific agonist being more potent than a naturally occurring RAR agonist. PMID- 11483255 TI - Identification of purinergic receptors in retinal ganglion cells. AB - P2X receptors are ligand-gated ion channels activated by adenosine triphosphate and expressed in a broad variety of tissues. The present study demonstrates the expression of various types of purinergic P2X receptors in identified retinal ganglion cells (RGCs) of the adult rat retina. Single-cell reverse transcription polymerase chain reaction (SC-RT-PCR) resulted in a positive amplification signal for all P2X receptor subunit mRNAs examined (P2X(3-5), P2X(7)). Immunohistochemistry with P2X(3,4) receptor subunit-specific antibodies showed a labelling of neurons in the ganglion cell layer and inner nuclear layer. Our data suggest that extracellular ATP acts directly on RGCs via several types of P2X receptors and may provide neuromodulatory influences on information processing in the retina. PMID- 11483256 TI - Proteomic analysis of proteins in PC12 cells before and after treatment with nerve growth factor: increased levels of a 43-kDa chromogranin B-derived fragment during neuronal differentiation. AB - Proteomic analysis is an important approach to characterizing the proteome and studying protein function in the post-genomic era. It is also a powerful screening method for detecting unexpected alterations in protein expression that may be missed by conventional biochemical techniques. The aim of this study was to perform a preliminary proteomic analysis of PC12 cells in order to investigate the effect of nerve growth factor (NGF) on protein expression in PC12 cells during neurite outgrowth. PC12 cell proteins were separated by two-dimensional electrophoresis (2DE) and visualized by silver staining, then certain proteins were identified by N-terminal amino acid microsequencing and a homology search of a protein sequence database. Over 400 proteins were detected, 10% of which showed a significant (greater than 30%) increase or decrease in expression during NGF induced neuronal differentiation. Seven proteins in the 2DE map were identified; the levels of five of these were unaffected by NGF treatment, whereas the levels of the other two, beta-tubulin and a novel 43-kDa chromogranin B-derived fragment, were significantly increased by more than 30 and 200%, respectively. Our results suggest that chromogranin B processing is enhanced in PC12 cells during NGF-induced neuronal differentiation. In addition, since this increase in the levels of the chromogranin B-derived fragment was specifically blocked by PD98059, we suggest that the increased processing can be ascribed to activation of the MAP kinase pathway, and that the 43-kDa chromogranin B-derived fragment can serve as a new marker of neuronal differentiation for proteomic studies. PMID- 11483257 TI - Sox8 gene expression identifies immature glial cells in developing cerebellum and cerebellar tumours. AB - Sox8 is a member of the E subgroup of Sox genes, the other members of which are Sox9 and Sox10, both of which are implicated in specific human disorders. Recently, Sox8 homologues have been cloned in chick, mouse and human and have been shown to be strongly expressed in the embryonic and adult brain. Nevertheless, the cell types that express Sox8 have not been determined. We show here that Sox8 is expressed in immature glia in the developing cerebellum. Sox8 is also expressed in scattered cells in the cerebellar tumour, medulloblastoma. This gene therefore provides an early glial marker that may provide more detailed insight into the cellular makeup and consequent behaviour of medulloblastomas. PMID- 11483258 TI - FMD in the UK--the 2001 outbreak; what if...? PMID- 11483259 TI - Endemic infection of a cat colony with a feline calicivirus closely related to an isolate used in live attenuated vaccines. AB - We have typed three feline calicivirus (FCV) isolates obtained over a 5-month period from an endemically infected cat colony. Sequence analysis from variable region E of the capsid gene from these isolates strongly suggests they are minor variants of a single FCV strain, and that this strain is closely related to the one used in many live-attenuated FCV vaccines. Such a vaccine was last used approximately 2 months before the first of the isolates in this study was obtained. Sequence differences between the 'colony isolate' and the vaccine virus suggest that the colony virus has evolved from the vaccine virus and was persisting in the colony. The extent to which vaccine virus may contribute to the continued high prevalence of FCV needs to be determined. PMID- 11483260 TI - Towards the rational design of Th1 adjuvants. AB - Finding adjuvants in order to enhance immune responses against target immunogens has been a major and recurrent issue for the vaccine industry. It is yet to be solved, most particularly in the context of a growing interest in designing new types of vaccines capable of eliciting Th1 immune responses. A review of synthetic adjuvants which have been (or are being) tested in clinical studies is presented. Importantly, recent advances in our understanding of the physiology of immune responses offer new avenues to design and test candidate adjuvants, based on either synthetic or natural molecules, with the aim to mimic and recapitulate pro-inflammatory signals initiating both innate and adaptative immune effector mechanisms. Thus, adjuvants of the future might be a mixture of molecules selected singularly for a capacity to attract, target or activate professional antigen presenting cells. Used as a combination, such molecules should facilitate antigen presentation by professional APCs and lead to a potent induction of T cell-mediated effector and immune memory mechanisms. PMID- 11483261 TI - Live attenuated influenza virus vaccines: new options for the prevention of influenza. AB - Live attenuated influenza virus (LAIV) vaccines present new possibilities for the prevention and control of influenza. Administered intranasally, LAIV vaccines offer a needle-free route of administration. These investigational vaccines have also been shown to be safe and effective in children and healthy working adults. A 2-year placebo-controlled trial among young children (1996-1997 and 1997-1998 influenza seasons) demonstrated that LAIV vaccine was associated with a 92% reduction in laboratory-confirmed cases of influenza. Vaccination also significantly reduced episodes of otitis media and antibiotic use. In a placebo controlled trial among healthy working adults during the 1997-1998 season, LAIV vaccine significantly reduced episodes of febrile upper respiratory tract illness and illness-associated work loss, health-care use, and antibiotic use. Seventy percent of study participants self-administered the vaccine. An economic analysis of the benefits of LAIV vaccine in this population suggests that the break-even cost for LAIV vaccine and its administration for healthy working adults would be about $39. For both children and healthy adults, LAIV vaccine provided substantial protection during the 1997-1998 season when the predominant circulating virus, the A/Sydney variant, was not contained in the vaccine. Studies are still underway to evaluate the potential incremental benefits of LAIV vaccine in addition to inactivated vaccine in high-risk populations. LAIV vaccines will be an important addition to the armamentarium for fighting influenza. PMID- 11483262 TI - Progress towards global polio eradication. AB - Twelve years after the global polio eradication goal was set, polio cases have declined by more than 95% world-wide. Polio immunization campaigns have been conducted in every endemic country with as many as 470 million children immunized per year. Intense wild poliovirus transmission is now limited to South Asia and sub-Saharan Africa. To achieve eradication at the earliest possible date, immunization campaigns are being intensified in the remaining endemic countries. Major programmatic challenges include reaching vulnerable children in areas with armed conflict and ensuring full financial and political support for the initiative. With global eradication imminent, WHO is preparing for post eradication issues: containment of polioviruses, certification of eradication, and stopping immunization. PMID- 11483263 TI - From genome to vaccine: in silico predictions, ex vivo verification. AB - Bioinformatics tools enable researchers to move rapidly from genome sequence to vaccine design. EpiMer and EpiMatrix are computer-driven pattern-matching algorithms that identify T cell epitopes. Conservatrix, BlastiMer, and Patent Blast permit the analysis of protein sequences for highly conserved regions, for homology with other known proteins, and for homology with previously patented epitopes, respectively. Two applications of these tools to epitope-driven vaccine design are described in this review. Using Conservatrix and EpiMatrix, we analyzed more than 10000 HIV-1 sequences and identified peptides that were potentially immunostimulatory and highly conserved across HIV-1 clades. MHC binding assays and CTL assays have been carried out: 50 (69%) of the 72 candidate epitopes bound in assays with cell lines expressing the corresponding MHC molecule; 15 of the 24 B7 peptides (63%) stimulated gamma-interferon release in ELISpot assays. These results lend support to the bioinformatics approach to selecting novel, conserved, HIV-1 CTL epitopes. EpiMatrix was also applied to the entire 'proteome' derived from two Mycobacterium tuberculosis (Mtb) genomes. Using EpiMatrix, BlastiMer, and Patent-Blast, we narrowed the list of putative Mtb epitopes to be tested in vitro from 1600000 to 3000, a 99.8% reduction. The pace of vaccine design will accelerate when these and other bioinformatics tools are systematically applied to whole genomes and used in combination with in vitro methods for screening and confirming epitopes. PMID- 11483264 TI - Immunogenicity and reactogenicity of diphtheria, tetanus and pertussis toxoids combined with inactivated polio vaccine, when administered concomitantly with or as a diluent for a Hib conjugate vaccine. AB - In an open trial, 400 infants were randomized to vaccination with a combined diphtheria-tetanus-pertussis-inactivated polio vaccine (DTaP-IPV) either mixed with a Haemophilus influenzae type b (Hib) tetanus toxoid conjugate immediately before injection (DTaP-IPV/Hib (mix)) or given concurrently with the Hib conjugate at separate injection sites (DTaP-IPV+Hib (sep)). The pertussis component consisted of pertussis toxoid alone. The vaccines were given intramuscularly at 3, 5 and 12 months of age. No vaccine-related serious adverse events occurred. Local reactions were evaluated from diary cards completed by the parents. Infants who received DTaP-IPV/Hib (mix) experienced fewer local reactions. Sera were obtained 28-45 days after the second and third vaccinations. Total Hib capsular antibodies were similar in the two groups after the second injection but lower in the group receiving DTaP-IPV/Hib (mix) than in the group receiving DTaP-IPV+Hib (sep) after the third injection (geometric mean 6.1 vs 10.4 microg/ml). Mixing of the vaccines also led to somewhat lower diphtheria toxin antibodies (5.9 vs. 7.7 IU/ml after the third injection) while tetanus antibodies were higher (3.9 vs. 2.5 IU/ml after the third injection). Antibodies against pertussis toxin and the three polio virus types were similar in the two groups. The moderate impairment of the Hib antibody response caused by mixing of the Hib conjugate with aluminium adsorbed DTaP may be due to physicochemical interference but is probably of little clinical importance because of the ability of the Hib conjugates to induce an immunologic memory. PMID- 11483265 TI - IgG antibody subclass responses determined by immunoblot in infants' sera following vaccination with a meningococcal recombinant hexavalent PorA OMV vaccine. AB - The introduction of meningococcal serogroup C conjugate vaccines into the UK immunisation schedule has led to the decline of serogroup C disease in those vaccinated but there is no imminent vaccine solution for serogroup B disease. The PorA outer membrane protein (OMP) is a potential serogroup B vaccine candidate and an outer membrane vesicle (OMV) vaccine containing six different PorA OMPs (each representing a different serosubtype) has been evaluated in phase II trials with encouraging results. Little is known about the IgG subclass response to the various antigens contained within this vaccine. These responses are important due to the different half-lives and complement fixing abilities of these antibodies. In this study, immunoblotting was undertaken with infants' sera following either three or four doses of vaccine, and OMVs from six isogenic meningococcal strains differing only in their PorA serosubtype. Following either three or four doses of the vaccine, IgG(3) and IgG(1) subclass antibodies were induced to all six of the isogenic strains, although sera collected after four doses of vaccine showed stronger antibody levels. IgG(3) was found in more sera than IgG(1). For both sets of sera, the two isogenic strains expressing P1.5,2 and P1.5(c),10 induced stronger IgG subclass antibody responses than the other four meningococcal strains. The recombinant hexavalent PorA OMV vaccine stimulates both IgG(1) and IgG(3) subclass antibodies, the subclasses that are most effective in activating the complement system. PMID- 11483266 TI - Passive transfer of protection against Bacillus anthracis infection in a murine model. AB - Passive transfer of lymphocytes and sera from mice immunised using two different formulations containing recombinant protective antigen (rPA) have been used to further elucidate the mechanism of protection against Bacillus anthracis infection. The results demonstrated that an antibody response maybe important in protection against B. anthracis infection, under the conditions tested. The results provide further data for the development of an improved anthrax vaccine. PMID- 11483267 TI - The comparative efficacy of CTLA-4 and L-selectin targeted DNA vaccines in mice and sheep. AB - The access of antigens to antigen presenting cells (APCs) appears to be a rate limiting step in the generation of immune responses to DNA vaccines. The cytotoxic T lymphocyte antigen 4 (CTLA-4) and L-selectin represent attractive ligands for use in the targeting of antigen to APCs and lymph nodes. CTLA-4 binds with high affinity to the B7 membrane antigen on APCs, while L-selectin functions as a lymphocyte homing marker and binds to CD34 on the surface of high endothelial venule cells. DNA vaccines encoding human immunoglobulin (HIg), fused to either CTLA-4 or L-selectin, have been shown to generate up to 10,000-fold higher anti-HIg antibody responses than DNA vaccines encoding HIg alone. In this study, the ability of CTLA-4 or L-selectin mediated targeting to enhance the humoral immune response to an alternate vaccine antigen was investigated. DNA vaccines encoding CTLA-4-HIg and L-selectin-HIg fused to the host-protective 45W antigen from Taenia ovis were constructed. In BALB/c mice, the L-selectin targeted vaccine did not improve either the magnitude or speed of antibody responses of vaccinated mice. In contrast, the CTLA-4 targeted DNA vaccine generated 45W-specific antibody responses which were up to 30-fold higher than those achieved with non-targeted DNA vaccination. The kinetic of the antibody response generated following CTLA-4 targeted DNA vaccination was also significantly faster than that achieved with non-targeted DNA vaccination, or with adjuvanted protein vaccination. Vaccination of outbred sheep with DNA vaccines expressing either murine or ovine CTLA-4 targeted antigen failed to enhance immune responses. These findings indicate that CTLA-4 targeting may find application in the improvement of DNA vaccines, but requires further development for applications in large animal species. PMID- 11483268 TI - A randomised comparison of two inactivated hepatitis A vaccines, Avaxim and Vaqta, given as a booster to subjects primed with Avaxim. AB - To investigate whether Vaqta may be used as a booster in subjects primed with Avaxim, 127 adults primed 6 months previously with Avaxim were randomised to receive either Vaqta or Avaxim as a booster. Prior to the booster all subjects were seropositive. Geometric mean antibody titres increased from 496 to 7262 mIU/mL 1 month after receiving Vaqta as a booster and from 325 to 5131 mIU/mL 1 month after receiving Avaxim as a booster. Both vaccines were well tolerated; 20.3% of subjects receiving Vaqta experienced a local reaction, compared to 39.7% of those receiving Avaxim. Systemic reactions were reported by 15.6% of those receiving Vaqta and 14.3% of those receiving Avaxim. Vaqta may be used as a booster in subjects primed with Avaxim. PMID- 11483269 TI - Protection against influenza virus challenge by topical application of influenza DNA vaccine. AB - We studied the use of a DNA vaccine expressing the matrix (M) gene of the influenza virus A/PR/8/34. Mice were immunized by painting the DNA vaccine three times on the skin after removal of its keratinocytic layers. Immunization by this method produced M-specific antibodies and cytotoxic T lymphocyte (CTL) response, and acquired resistance against influenza virus challenge. This protection was abrogated by the in vivo injection of anti-CD8 or anti-CD4 monoclonal antibody. We further found that simultaneous topical application (t.a.) of GM-CSF expression plasmid (pGM-CSF) or liposomes plus mannan produced stronger immune response competence and enhanced the protective effect against influenza virus challenge. The present study revealed that administering DNA vaccine by topical application can elicit both humoral and cell-mediated immunity (CMI). PMID- 11483270 TI - Biodegradable PLGA microspheres as a delivery system for malaria synthetic peptide SPf66. AB - SPf66 is the first chemically synthesised vaccine to elicit a partial protective immune response against malaria. The aluminium hydroxide (alum)-adsorbed SPf66 vaccine is weakly immunogenic and of poor to moderate efficacy in humans. To investigate the possibility of improving SPf66 vaccine immunogenicity, a delivery system based on poly-D,L-lactide-co-glycolide (PLGA) microspheres was developed and the immune response induced after its subcutaneous administration into mice was evaluated. Microspheres were prepared by a solvent extraction/double emulsion (w/o/w) method and characterised for morphology, size, peptide loading, release profile and peptide integrity. The in vitro and in vivo results obtained showed that there was no apparent effect of the encapsulation procedure on SPf66 integrity and immunogenicity. The subcutaneous administration of microspheres showed a significantly higher immune response (serum IgG levels) than that obtained with alum adsorbed SPf66 and it was comparable to that of SPf66 emulsified with Freund's adjuvant (FA). These observations illustrate the potential of PLGA microspheres as a delivery system for chemically synthesised antigens. PMID- 11483271 TI - Antigenic and molecular heterogeneity in recent swine influenza A(H1N1) virus isolates with possible implications for vaccination policy. AB - In order to explore the occurrence of antigenic drift in swine influenza A(H1N1) viruses and the match between epidemic and vaccine strains, 26 virus isolates from outbreaks of respiratory disease among finishing pigs in the Netherlands in the 1995/1996 season and reference strains from earlier outbreaks were examined using serological and molecular methods. In contrast to swine H3N2 viruses, no significant antigenic drift was observed in swine H1N1 viruses isolated from the late 1980s up to 1996 inclusive. However, a marked antigenic and genetic heterogeneity in haemagglutination inhibition tests and nucleotide sequence analyses was detected among the 26 recent swine H1N1 virus strains. Interestingly, the observed antigenic and molecular variants were not randomly distributed over the farms. This finding indicates independent introductions of different swine H1N1 virus variants at the various farms of the study and points to a marked difference between the epidemiologies of human and swine influenza viruses. The observed heterogeneity may hamper the control of swine influenza by vaccination and indicates that the efficacy of current swine influenza vaccines requires re-evaluation and that the antigenic reactivity of swine influenza viruses should be monitored on a regular basis. PMID- 11483272 TI - Role of antibody to lipopolysaccharide in protection against low- and high virulence strains of Francisella tularensis. AB - Mice immunised with lipopolysaccharide (LPS) from Francisella tularensis were protected against challenge with the live vaccine strain (LVS). However, when similarly immunised mice were challenged using the fully virulent F. tularensis strain Schu4, only an increase in the time to death was observed. Passive transfer of serum from LPS-immunised mice to naive mice afforded protection against F. tularensis LVS. LPS-immunised mice depleted of either CD4+ or CD8+ T cells survived a F. tularensis LVS challenge although the rate of clearance of bacteria from the spleen was significantly reduced in the CD8+ depleted group. LPS-immunised mice boosted with F. tularensis LVS were re-challenged with F. tularensis Schu4. This cohort was significantly protected (LD(50) increased from <1 to >1000 CFU). However, passive transfer of serum did not confer protection and mice depleted of CD4+ or CD8+ T-cells did not survive. PMID- 11483273 TI - Immunogenicity and efficacy of one dose measles-mumps-rubella (MMR) vaccine at twelve months of age as compared to monovalent measles vaccination at nine months followed by MMR revaccination at fifteen months of age. AB - BACKGROUND AND METHODS: measles is a common cause of morbidity and mortality in developing countries. Although the measles-mumps-rubella vaccine (MMR) is currently in use in developed countries, monovalent measles vaccine (MV) is routinely recommended by World Health Organization (WHO) at 9 months of age in Turkey, as in many other developing countries. In this study, 442 Turkish children received MV at 9 months of age and were revaccinated with MMR vaccine at 15 months of age. In the second group 495 children received MMR at 12 months of age with no earlier measles vaccination. Antibodies were measured before the first vaccination and 6 weeks after the MMR. All children had been followed for occurrence of measles infection for 60 months. Two vaccination schedules were compared for immunogenicity and protection rates. CONCLUSIONS: seroconversion and clinical protection rates were significantly higher in children who received only MMR at 12 months of age than in children revaccinated at 15 months of age. Seroconversion rate for measles was 69.9% in children who received MMR at 12 months of age and 90.3% in children revaccinated at 15 months of age (P=0.0003). While there was no measles case in children who were revaccinated, 12 (2.7%) children in the first group acquired measles during the follow-up period. Vaccination at 12 months of age appeared to be better than the current national standard. The late elimination of maternal antibodies and the inhibitory effect of a weak antibody response after the first dose of vaccine at 9 months may explain the better immunogenicity and efficacy of the MMR vaccine given at 12 months of age. PMID- 11483274 TI - Efficacy of vaccination of pigs with different H1N1 swine influenza viruses using a recent challenge strain and different parameters of protection. AB - This study investigates whether antigenic evolution within H1N1 swine influenza viruses can compromise vaccine efficacy and, specifically, whether the A/New Jersey/8/76 strain in the commercial swine influenza vaccines needs to be updated. Pigs were vaccinated twice intramuscularly with experimental monovalent vaccines derived from different H1N1 strains (A/New Jersey/8/76, Sw/Belgium/1/83 or Sw/Belgium/1/98) or with a commercial bivalent vaccine based on A/New Jersey/8/76 (H1N1) and A/Port Chalmers/1/73 (H3N2). Experimental and commercial vaccines contained a different adjuvant. Two weeks after the second vaccination, all pigs were challenged intratracheally with Sw/Belgium/1/98. Mean pre-challenge haemagglutination inhibition (HI) antibody titres against the challenge virus were lower for the experimental A/New Jersey/8/76 vaccine than for the other vaccines. The reduction in mean virus titres in the lungs was highly significant for the latter vaccines, including the commercial New Jersey-derived vaccine, but not for the experimental A/New Jersey/8/76 vaccine. Clinical signs after challenge were negligible in all vaccinates. Post-challenge levels of interferon alpha and tumor necrosis factor-alpha in bronchoalveolar lavage fluids were reduced in the vaccinates, while levels of interleukin-1 and neutrophils were less consistent. Though the A/New Jersey/8/76 strain is less effective in preventing infection by Sw/Belgium/1/98 than the homologous virus or than Sw/Belgium/1/83, all strains can protect completely if antibody titres against the challenge virus are sufficiently high. Apart from the vaccine strain, adjuvant and antigenic dose may play a crucial role in vaccine efficacy. PMID- 11483275 TI - Plasmodium falciparum circumsporozoite (CS) protein peptides specifically bind to HepG2 cells. AB - Hepatocyte invasion by malaria parasites is mediated by specific molecular interactions. Several lines of evidence suggest the importance of the surface plasmodial circumsporozoite (CS) protein in the sporozoite invasion of hepatocytes. Identification of the sequences involved in binding to hepatocytes is an important step towards understanding the structural basis for the sporozoite-hepatocyte interaction. In this study, binding assays between Plasmodium falciparum CS peptides and HepG2 cells were performed. Fifteen overlapping residue 20 mer long peptides, spanning the entire CS sequence, were tested in HepG2 cell binding assays. Five High Binding Activity Peptides (HBAPs) to HepG2 cells were identified: 4593, (NANPNANPNANP); 4383, (NSRSLGENDDGNNEDNEKLR); 4388, (GNGQGHNMPNDPNRNVDENA); 4389, (HNMPNDPNRNVDENANANSA) and 4390, (DPNRNVDENANANSAVKNNN). The HBAP HepG2 interaction is independent of charge and amino-acid composition, but sequence dependent. Four HBAPs (4383, 4388, 4389 and 4390) are bound with similar affinity to a 50 kDa molecule. These HBAPs define three Hepatocyte Binding Sequences (HBSs): HBS-1, located between residues 68 and 87 (HBAP 4383); HBS-11, the repeat NANP region (HBAP 4593), for which anti repeat antibodies are able to specifically inhibit sporozoite invasion of hepatocytes have been reported; and HBS-111, between residues 286 and 315 (HBAPs 4388, 4388 and 4390), respectively. Interestingly, HBS 111 carries two earlier-reported B-epitopes (underlined) in peptides 4388, 4389 and 4390 (GNGQGHNMPNDPNRNVD ENANANSAVKNN) in its sequence. The HBSs reported here show lesser interspecie-variability than the entire protein in species invading the same kind of hepatic cells. This data supports these HBSs' important role in CS-protein function; they could be used as ligand by the sporozoite to invade hepatic cells. PMID- 11483276 TI - Plasmodium falciparum acid basic repeat antigen (ABRA) peptides: erythrocyte binding and biological activity. AB - Non overlapping 20-mer peptides, covering the complete sequence of acid basic repeat antigen (ABRA) of Plasmodium falciparum, were synthesised and tested in binding assays to erythrocytes. Five peptides localised in the N-terminal region coded 2148 (121LQSHKKLIKALKKNIESYQN(140)), 2149 (141KKHLIYKNKSYNPLLLSCVK(160)), 2150 (161KMNMLKENVDYIQKNQNLFK(180)), 2152 (201YKSQGHKKETSQNQNENNDN(220)) and 2153 (221QKYQEVNDEDDVNDEEDTND(240)) specifically bind to erythrocytes. These peptides bind independently of the peptide and erythrocyte charge, with high affinity (Kd between 70 and 180 nM) and the hydrophobic interaction is important for this binding ( approximately 30% hydrophobic critical residues). These results allow us define a specific erythrocyte binding region (residues 121-240), which may bound to at least three different binding sites on erythrocytes. Peptide 2153 shares the underlined sequence 221QKYQEVNDEDDVNDEEDTND(240) with an earlier 18 mer peptide recognised by human exposed sera. Peptides number 2148 and 2149 in vitro inhibit erythrocyte invasion by merozoites. We found that 2149 peptide and some of its glycine analogues show specific haemolytic and/or antimicrobial activity. We discuss a possible role of ABRA or its regions in the merozoite invasion of erythrocyte. PMID- 11483277 TI - Potential of Klebsiella pneumoniae cytotoxin toxoid as vaccine against klebsiellosis in rabbits and mice. AB - The protective efficacy of toxoids prepared from crude cytotoxin (polymyxin-B extract, PBE) and purified Klebsiella cytotoxins (KCTs) was studied in rabbits and mice. The toxoids of KCT-I and PBE were found to be protective against homologous as well as heterologous Klebsiella challenges, while toxoids of KCT-II and KCT-III afforded protection in mice against homologous Klebsiella infections. KCT-I and PBE toxoids also induced good humoral anti-Klebsiella response in rabbits with ELISA titres ranging from 20480 to 81480. Immunized female rabbits passed protective anti-Klebsiella immunoglobulins to their offsprings through colostra. Baby rabbits fed on colostrum of immunized rabbits withstood lethal Klebsiella infection up to 1 month of age, but not on the 50th day. Baby rabbits having an anti-Klebsiella IgG titre > or =1280 were fully protected against lethal dose of Klebsiella. The study revealed a significant protective efficacy of KCT-I and PBE toxoids against klebsiellosis in mice and rabbits. PMID- 11483278 TI - Rapid clearance of SAG-2 rabies virus from dogs after oral vaccination. AB - This study investigated the safety, efficacy, and clearance of SAG-2, an attentuated rabies virus, after oral vaccination in dogs. Nineteen dogs consumed baits containing lyophilized vaccine, but residual SAG-2 virus was recovered in only one of 57 oral swabs, collected one hour post-vaccination. Seven vaccinates were euthanized between 24 and 96 h after consuming a bait. Rabies virus RNA was detected in tonsils from all seven dogs by nested RT-PCR, with primers to the viral glycoprotein. Genomic, sense-transcripts, and m-RNAs were detected in five of seven tonsil samples using primers to the rabies virus nucleoprotein gene, as well as in four of seven samples from the buccal mucosa and one of seven from the tongue. Rabies virus antigen was detected in all tonsils by an immunohistochemistry test, confirming the RT-PCR results. In addition, virus was isolated from one tonsil sample collected at 96 h, providing supportive evidence of viral replication. Ten of 12 (83%) of the vaccinated dogs demonstrated an anamnestic response, with viral neutralizing antibody titers (> or =0.5 IU/ml), after rabies virus challenge. These ten dogs survived, whereas all control dogs succumbed to rabies. Attenuated rabies viruses, such as SAG-2, replicate in local tissues of the oral cavity and can be cleared relatively quickly, without viral excretion, leading to protective immunity against the disease. PMID- 11483279 TI - Nasal absorption and biodistribution of plasmid DNA: an alternative route of DNA vaccine delivery. AB - Nasal administration is emerging as a new route of DNA vaccine delivery. We aimed to study the extent of absorption and biodistribution of intranasally administered plasmid DNA. After intranasal administration, the level of plasmid DNA in the serum peaked at 1.5 h. The ratio of the area under the concentration (AUC) after intranasal administration of DNA over the AUC after intravenous administration was 0.14. At 15 min post inoculation, the highest organ distribution was observed in the liver and the cervical lymph nodes showed the highest level among the lymph nodes. At 24 h a higher localization of plasmids to the brain than to the lung and spleen was notable. A significant level of mRNA expression was observed in the lymph nodes. These results suggest that plasmid DNA can be substantially absorbed and distributed to the lymph nodes after intranasal administration, partly explaining the systemic immunogenicity of intranasally administered plasmid DNA vaccines. PMID- 11483280 TI - DNA vaccination against v-src oncogene-induced tumours in congenic chickens. AB - DNA vaccination is particularly efficient for induction of cytotoxic T-lymphocyte (CTL) response. In our experiments, we used MHC(B) congenic chicken lines CB and CC (regressors and progressors of v-src-induced tumours, respectively) and a mutated, non-oncogenic v-src gene construct as the DNA vaccine. A high degree of vaccine protection against oncogenic v-src challenge was achieved in the CB line chickens. CTL response was demonstrated in vitro and by adoptive transfer of immune cells to the syngeneic host and to the CC line chickens rendered tolerant to CB cells. In the CC line chickens we observed tumour growth retardation after a low-dose DNA vaccination administered to immature recipients while higher amounts of DNA vaccine in immunocompetent chickens exerted an enhancing effect. PMID- 11483281 TI - First field trial of a transmissible recombinant vaccine against myxomatosis and rabbit hemorrhagic disease. AB - As a novel approach for immunisation of wild rabbits, we have recently developed a transmissible vaccine against myxomatosis and rabbit hemorrhagic disease (RHD) based on a recombinant myxoma virus (MV) expressing the RHDV capsid protein [J. Virol. 74 (2000) 1114]. The efficacy and safety of the vaccine have been extensively evaluated under laboratory conditions. In this study, we report the first limited field trial of the candidate vaccine that was undertaken in an island of 34 Has containing a population of around 300 rabbits. Following administration by the subcutaneous route to 76 rabbits, the vaccine induced specific antibody responses against both myxomatosis and RHDV in all the inoculated rabbits. Furthermore, the recombinant virus exhibited a limited horizontal transmission capacity, promoting seroconversion of around 50% of the uninoculated rabbit population. No evidence of undesirable effects due to the recombinant virus field release was detected. PMID- 11483282 TI - High seroprotection rate induced by low doses of a recombinant hepatitis B vaccine in healthy Iranian neonates. AB - Low-dose vaccination has been proposed as a cost-saving strategy to implement mass vaccination of neonates with hepatitis B (HB) vaccine, world-wide, particularly in developing countries. The effectiveness of low vaccine doses, however, needs to be evaluated and verified in newborns from different ethnic backgrounds. In this study, a recombinant HB vaccine (Heberbiovac) was administered in three different doses, including the standard 10 microg together with 5 and 2.5 microg to three groups of healthy Iranian neonates. Each group, consisting of 230-260 subjects, received triple doses of vaccine given i.m. at 0, 1.5 and 9 month intervals. Serum samples were collected 2-4 weeks after completion of vaccination, and anti-HBs antibody was quantitated by a sandwich ELISA. Seroprotection (anti-HBs>10 IU/L) was detected in 92, 97.4 and 96.1% of 2.5, 5 and 10 microg vaccine dose recipients, with geometric mean titer (GMT) of 4502, 5824 and 6104 IU/L, respectively. No significant differences were observed in seroprotection rate and GMT between the 10 and 5 microg dose recipients. Both parameters, however, were significantly lower in neonates vaccinated with a 2.5 microg vaccine dose (P<0.05 and P<0.001, respectively). These findings indicate induction of a strong protective immune response in healthy Iranian neonates by a low dose (5 microg) of the recombinant HB vaccine. PMID- 11483283 TI - Features of the antibody response attributable to plasmid backbone adjuvanticity after DNA immunization. AB - DNA vaccination induces antigen-specific immune responses with characteristics distinct from other vaccination modes. In the present study, the contribution of the plasmid backbone adjuvant effect to the quality of the DNA-raised antibody response was investigated. For this purpose, three intradermal primings were compared in mice using: (1) the recombinant Schistosoma haematobium glutathione S transferase antigen (rSh28GST): (2) rSh28GST supplemented with a non-coding plasmid; and (3) a Sh28GST-encoding plasmid. In contrast to immunization with the protein, DNA immunization elicited a very stable antibody (Ab) response over a prolonged period of time. This feature was attributed to the plasmid backbone, because co-administration of the non-coding plasmid with rSh28GST allowed the maintenance of the specific Ab response. A strong anamnestic Ab response was induced after intradermal boost with rSh28GST only in the mice primed with pMSh. This indicated that the selective ability of DNA vaccination to induce memory humoral response was independent of the plasmid backbone. In contrast the plasmid backbone was found to strongly participate in the preferential IgG2a Ab production observed. These results suggest that, following DNA immunization, the Th1-biased profile and the maintenance of the long-lived Ab response could be attributed to an adjuvant effect of the plasmid backbone during priming, whereas the strength of B-cell memory was independent of this effect. PMID- 11483284 TI - A purified inactivated Japanese encephalitis virus vaccine made in Vero cells. AB - A second generation, purified, inactivated vaccine (PIV) against Japanese encephalitis (JE) virus was produced and tested in mice where it was found to be highly immunogenic and protective. The JE-PIV was made from an attenuated strain of JE virus propagated in certified Vero cells, purified, and inactivated with formalin. Its manufacture followed current GMP guidelines for the production of biologicals. The manufacturing process was efficient in generating a high yield of virus, essentially free of contaminating host cell proteins and nucleic acids. The PIV was formulated with aluminum hydroxide and administered to mice by subcutaneous inoculation. Vaccinated animals developed high-titered JE virus neutralizing antibodies in a dose dependent fashion after two injections. The vaccine protected mice against morbidity and mortality after challenge with live, virulent, JE virus. Compared with the existing licensed mouse brain-derived vaccine, JE-Vax, the Vero cell-derived JE-PIV was more immunogenic and as effective as preventing encephalitis in mice. The JE-PIV is currently being tested for safety and immunogenicity in volunteers. PMID- 11483285 TI - Surveillance for meningococcal disease and strategies for use of conjugate meningococcal vaccines in the United States. AB - BACKGROUND: Neisseria meningitidis is a leading cause of bacterial meningitis in US; new capsular type-specific conjugate vaccines offer an opportunity for improved control of meningococcal disease. We evaluated the relative burdens of invasive meningococcal disease in US and examined the projected impact of various meningococcal conjugate vaccination strategies on rates of meningococcal disease. METHODS: Meningococcal disease incidence rates were determined from active, population-based surveillance in selected US areas. Models were created to determine impact of vaccination of infants, toddlers, adolescents or college students with meningococcal conjugate vaccines, with assumptions for vaccine coverage, efficacy and duration of protection. Although we examined possible conjugate vaccine formulations including serogroups A, C, Y and W-135, the final vaccine impact analysis excluded serogroups A and W-135. Outcome measures were cumulative meningococcal disease incidence, and incidence 10 years after initiating vaccination among 0-22-year-olds. RESULTS: In models of serogroup C+Y meningococcal conjugate vaccination of infants, toddlers and adolescents, the cumulative incidence of meningococcal disease was reduced by 54, 48 and 25%, respectively; the toddler strategy had the greatest impact per dose. After 10 years of routine meningococcal conjugate vaccination, meningococcal disease could be reduced by 50% and deaths by 64%. CONCLUSIONS: Use of meningococcal conjugate vaccine could markedly reduce meningococcal disease incidence. Our data, along with vaccine formulation and vaccination program considerations, will be important in determining the optimal choice of vaccination strategy. PMID- 11483286 TI - Effect of acute and subchronic subcutaneous urocortin on blood pressure and food consumption in ob/ob mice. AB - Corticotropin-releasing factor and urocortin belong to a hypothalamic peptide family thought to be important in appetite regulation. The present study compared the appetite-suppressant effect of subcutaneous urocortin in obese mice to its cardiovascular effects. Acutely, urocortin (100 nmol/kg iv) reduced blood pressure and increased heart rate in urethane anesthetized nonobese mice; effects similar to those produced by subcutaneous urocortin (10 and 100 nmol/kg sc) in nonobese and ob/ob mice. Over this same dose range (10-100 nmol/kg sc), urocortin dramatically inhibited food consumption in the ob/ob mouse. To determine if the acute hypotensive effect of urocortin (10 nmol/kg sc) in the ob/ob mouse persisted after repeated urocortin administration, animals were pretreated for 3 days with urocortin (10 nmol/kg sc) or vehicle. Following urocortin pretreatment, urocortin-induced hypotension was similar to the effect in vehicle pretreated mice. However, urocortin-induced appetite suppression was reduced following 3 days of pretreatment with urocortin (10 nmol/kg sc) to ob/ob mice. These data suggest that the hypotensive and appetite-suppressant effects of urocortin are mediated by different mechanisms and tolerance to the hypotension did not readily occur in obese animals. PMID- 11483287 TI - Impaired relaxation to acetylcholine in 2K-1C hypertensive rat aortas involves changes in membrane hyperpolarization instead of an abnormal contribution of endothelial factors. AB - The contribution of endothelial factors and mechanisms underlying decreased acetylcholine-induced relaxation and endothelial inhibitory action on phenylephrine-induced contraction were evaluated in aortas of two-kidney, one clip hypertensive (2K-1C) and normotensive (2K) rats. Relaxation induced by acetylcholine in 2K-1C precontracted by phenylephrine was lower [Maximum Effect (ME): 71.33+/-3.36%; pD(2): 7.050+/-0.03] than in 2K (ME: 95.26+/-1.59%; pD(2): 7.31+/-0.07). This response was abolished by N(G)-nitro-L-arginine (L-NNA) in 2K 1C, but was only reduced in 2K (ME: 29.21+/-9.28%). Indomethacin had no effect in 2K-1C, and slightly attenuated acetylcholine-induced relaxation in 2K. The combination of L-NNA and indomethacin almost abolished acetylcholine-induced relaxation in 2K-1C, while in 2K, the inhibition (ME: 56.61+/-8.95%) was lower than the effect of L-NNA alone. During the KCl-induced precontraction, 2K and 2K 1C aortas showed similar acetylcholine-induced relaxation (43.50+/-5.64% vs. 41.60+/-4.36%), which was abolished by L-NNA. The levels of cGMP produced in response to acetylcholine were not different between 2K and 2K-1C. The sensitivity to sodium nitroprusside was lower in phenylephrine-precontracted aortas from 2K-1C than 2K, as showed by the pD(2) values (7.72+/-0.20 vs. 8.59+/ 0.17), and this difference was abolished in aortas precontracted by KCl. The membrane potential was less negative in 2K-1C than in 2K (-41.57+/-1.19 vs. 51.00+/-1.13 mV) and hyperpolarization induced by acetylcholine was lower in 2K 1C than in 2K aortas (6.00+/-0.66 vs. 13.27+/-1.61 mV). Phenylephrine-induced contraction in aortas with endothelium was similar in both groups, and increased by the endothelium removal. This increase was lower in 2K-1C (from 1.32+/-0.06 to 1.90+/-0.21 g) than 2K (from 1.49+/-0.07 to 2.83+/-0.18 g). L-NNA and the endothelium removal had similar effect in 2K-1C (1.85+/-0.18 g) and were lower in 2K (2.18+/-0.20 g). Indomethacin decreased phenylephrine-induced contraction only in 2K. In conclusion, our major finding was a selective defect in smooth muscle membrane hyperpolarization, which could explain the decreased relaxation to acetylcholine and the attenuated inhibitory effect of endothelium on the contractile function in 2K-1C aortas. PMID- 11483288 TI - Pharmacological effects of an aldehyde type alpha/beta-adrenoceptor blocking agent with vasodilating properties. AB - KMUP 880723 (0.5, 1.0, and 3.0 mg/kg, iv) produced dose-dependent hypotensive and bradycardia responses in pentobarbital-anesthetized Wistar rats. KMUP 880723 (1.0 mg/kg, iv) also markedly inhibited both the tachycardia effects induced by ( )isoproterenol and arterial pressor responses induced by phenylephrine. In the isolated Wistar rat right atria, left atria, and guinea pig tracheal strips, KMUP 880723 competitively antagonized the (-)isoproterenol-induced positive chronotropic effects, inotropic effects, and tracheal relaxation effects in a concentration-dependent manner. The parallel shift to the right of the concentration-response curve of (-)isoproterenol suggested that KMUP 880723 was a beta(1)/beta(2)-adrenoceptor competitive antagonist. The apparent pA(2) values were 6.89+/-0.10 in the right atria, 7.02+/-0.09 in the left atria, and 6.59+/ 0.11 in the trachea, indicating that KMUP 880723 was a nonselective beta adrenoceptor blocker. In thoracic aorta experiments, KMUP 880723 also produced a competitive antagonism of norepinephrine-induced contraction with a pA(2) value of 7.14+/-0.06. In isolated rat thoracic aorta, KMUP 880723 more potently relaxed the contractions induced by norepinephrine (3 x 10(-6) M) than those by high K(+) (75 mM). In the radioligand-binding assay, the pK(i) values of [3H]CGP-12177 binding to rat ventricle and lung membranes were 6.56 and 6.40, respectively, and the value of [3H]prazosin binding to rat brain membranes was 6.66. These results further confirmed the alpha/beta-adrenoceptor blocking activities of KMUP 880723 reported in the functional studies. We conclude that KMUP 880723 is a nonselective beta-adrenoceptor antagonist with alpha-adrenoceptor blocking associated vasorelaxant activity. PMID- 11483289 TI - Effect of nitrite on endothelial function in isolated lung. AB - Nitrated tyrosine, implicated in protein dysfunction, is increased in various tissues in association with diverse pathological processes. Angiotensin converting enzyme (ACE) is a luminal vascular endothelial enzyme whose dysfunction is an early sign of endothelial injury. ACE contains a tyrosine critical for its enzymatic activity. Others have shown that nitrite exacerbates the ACE dysfunction of cultured endothelial cells in contact with activated polymorphonuclear neutrophils (PMN). We hypothesized that exogenous nitrite would enhance endothelial ACE dysfunction associated with PMN activation in the isolated lung. Rats received lipopolysaccharide (LPS) 2 h prior to isolated lung perfusion with Ficoll containing buffer. Either formyl-Met-Leu-Phe (fMLP, 10(-7) M) or phorbol myristate acetate (PMA, 10(-7) M) was used to activate PMN in lungs treated or not treated with 300-microM nitrite. A first pass indicator dilution method and first order reaction kinetics were used to determine ACE activity, while lung Ficoll content served as an index of vascular permeability. Both fMLP and PMA decreased endothelial ACE activity and increased pulmonary artery pressure, edema and vascular permeability. Exogenous nitrate did not potentiate the decrease in ACE activity, the lung injury or nitrotyrosine immunoreactivity of lung homogenates. In contrast to observations in cultured endothelial cells, our findings in the whole lung are compatible with the speculation of others that the rat lung has an unidentified factor, which minimizes accumulation of nitrated proteins. PMID- 11483290 TI - Postglomerular vasoconstriction to angiotensin II and norepinephrine depends on intracellular calcium release. AB - The current study was performed to determine the effect of calcium store depletion with cyclopiazonic acid (CPA) on the pre- and postglomerular vasoconstrictor responses to angiotensin II (ANG II) and norepinephrine (NE). CPA treatment significantly attenuated the afferent arteriolar response to 10 nM ANG II by 51% and to 1000 nM NE by 19%. Efferent arteriolar responses to ANG II and NE were also greatly attenuated in the presence of CPA. These data demonstrate that afferent and efferent arteriolar responses to ANG II and NE depend on release of calcium from CPA-sensitive intracellular stores. Furthermore, the postglomerular response to these agents exhibits a greater dependency on calcium release from intracellular stores. PMID- 11483291 TI - Spiradoline, a kappa opioid receptor agonist, produces tonic- and use-dependent block of sodium channels expressed in Xenopus oocytes. AB - Spiradoline, an arylacetamide kappa (kappa) opioid receptor agonist, produced a potent tonic block of rat neuronal (EC(50)= 34+/-5 microM) and heart (EC(50)= 183+/-13 microM) sodium channels and also blocked IFMQ3 mutant neuronal sodium channels (EC(50)= 130+/-34 microM) that lack fast inactivation when expressed in Xenopus oocytes. Spiradoline produced a hyperpolarizing shift in the voltage dependence of sodium channel inactivation and exhibited a marked frequency dependent component to blockade of sodium channels. The onset of open channel block of the IFMQ3 channel by spiradoline was best fit with a first-order blocking scheme, yielding an affinity constant of 116 +/- 33 microM. Thus, spiradoline blocks sodium channels by interacting with the major states of the channel which could result in local anesthetic action in nerves and antiarrhythmic action in the heart. PMID- 11483292 TI - Changes in vascular reactivity induced by acute hyperthyroidism in isolated rat aortae. AB - Hyperthyroidism was induced by subcutaneous injections of L-thyroxine (T(4)) (500 mg/kg/day) for 3 days in order to study whether adrenergic and muscarinic receptor-mediated vascular responses alter at an early stage of the disease. T(4) treatment was sufficient to induce a significant degree of thyroid weight loss, tachycardia, cardiac hypertrophy, and an elevation in serum T(4) levels. The tension of aortic ring preparations isolated from rats was measured isometrically to investigate the influence of acute hyperthyroidism. The contractions induced by norepinephrine (NE) were significantly suppressed in aortic rings from rats treated with T(4) compared with control rats. N(G)-nitro-L-arginine (L-NOARG), an inhibitor of nitric oxide synthase (NOS), significantly enhanced NE-induced contraction in aortic rings from both control and T(4)-treated rats, and the enhancement was greater in rats treated with T(4) than control rats. The relaxations induced by either acetylcholine (ACh) or sodium nitroprusside (SNP) were also significantly enhanced by T(4) treatment. L-NOARG abolished the relaxation induced by ACh in aortic rings from both control and T(4)-treated rats. L-NOARG shifted SNP-induced relaxation curves of aortic rings from those of control rats to the left, but not with rats treated with T(4). T(4) treatment showed no influence on the amount of endothelial NOS (eNOS) protein. These results suggest that vascular responses alter at an early stage of hyperthyroidism and that it may be due to a modification in the NO system which is independent from the amount of eNOS protein. PMID- 11483293 TI - Relaxation induced by histamine is not endothelium dependent in tail arteries of L-NAME-treated rats. AB - The present study was carried out to evaluate the relaxation induced by histamine in tail arteries of rats after chronic inhibition of nitric oxide (NO) synthesis with the inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) compared to tail arteries of control rats. The maximum relaxation induced by histamine was greater in control (88.09% +/-5.50, n=6) than in L-NAME arteries (47.33% +/-6.40, n=6), although pD(2) values were not different between the two groups (control: 4.89+/ 0.08; L-NAME: 4.81+/-0.10). After incubation with 100 microM L-NAME in vitro, the maximum relaxation induced by histamine was only reduced in the control arteries (44.93% +/-2.35, n=6), whereas it had no effect on aortas of rats pretreated with this inhibitor. The incubation with 100 microM L-NAME had the same effect as endothelium removal in both arterial groups. Furthermore, the relaxation induced by histamine was unaffected by indomethacin. The combination of L-NAME and the histamine antagonist cimetidine completely abolished the relaxation induced by histamine in both arterial groups. These results show that when NO synthesis is impaired, the relaxation induced by histamine is endothelium independent, and when NO-synthase is active, the relaxation involves both NO released from endothelial cells and an endothelium-independent mechanism that is sensitive to cimetidine. PMID- 11483295 TI - Damage to the vestibular inner ear causes long-term changes in neuronal nitric oxide synthase expression in the rat hippocampus. AB - The vestibular inner ear detects head acceleration and initiates compensatory eye movement and postural reflexes that help keep the visual image of the world stable on the retina, and maintain balance, during unexpected head movement. The most primitive vestibular systems are estimated to have evolved more than 500 million years ago and in mammalian and submammalian species the vestibular reflexes are mediated by basic brainstem pathways (see Wilson and Melvill Jones, 1979 for review). Although the contributions of the vestibular system to higher cognitive function have generally received less attention than its reflexive roles, vestibular sensory information is transmitted to higher centres in the brain and humans with vestibular damage are known to experience debilitating perceptual illusions (see Curthoys and Halmagyi, 1995; Berthoz, 1996 for reviews). Increasing behavioural and neurophysiological evidence suggests that the hippocampus uses information from the vestibular inner ear in order to build up maps of space that can be used in the development of spatial memory during learning tasks (McNaughton et al., 1991; Chapuis et al., 1992; Wiener and Berthoz, 1993; O'Mara et al., 1994; Wiener et al., 1995; Gavrilov et al., 1995; Stackman and Taube, 1996; Vitte et al., 1996; Taube et al., 1996; Save et al., 1998; Peruch et al., 1999; Cuthbert et al., 2000; Russell et al., 2000). However, to date, there has been no indication of the long-term neurochemical effects of the loss of vestibular input on hippocampal function. Since nitric oxide has been implicated in the mechanisms of hippocampal synaptic plasticity associated with the development of short-term memory (e.g. Schuman and Madison, 1991; Schuman et al., 1994; Arancio et al., 1996; Wu et al., 1997; Lu et al., 1999), we examined whether changes occur in the activity and expression of the enzymes responsible for nitric oxide production (nitric oxide synthases) in subregions of the rat hippocampus at different times following unilateral peripheral vestibular lesions, using western blotting and radioenzymatic assays. We found a decreased expression of neuronal nitric oxide synthase in the ipsilateral dentate gyrus at 2 weeks following the vestibular damage and not before, that may be related to the long-term effects of the loss of vestibular input on hippocampal function. These results support the hypothesis that head movement and position information derived from the vestibular inner ear may be important for the normal function of the hippocampus. PMID- 11483296 TI - Translating developmental time across mammalian species. AB - Conservation of the order in which events occur in developing mammalian brains permits use of regression theory to model the timing of neural development. Following a small adjustment to account for a systematic variability in primate cortical and limbic systems, the model is used to generate a 95-event/nine species matrix that predicts aspects of neurogenesis and axonal outgrowth in the brains of developing mice, hamsters, rats, spiny mice, rabbits, ferrets, cats, monkeys, and humans. Although data are compiled from species in which the timing of birth and the rate of maturation vary widely, the model proves statistically accurate, with practical implications for improving estimation of milestones of neural development, particularly for humans. Using the three-factor model (species, neural events, and primate adjustments), we produce predictions for the timing of 493 neural occurrences in developing mammalian brains that either have not yet been, or cannot be, empirically derived. We also relate the timing of neural events across the nine species in the form of a reference table calibrated to the development of laboratory rats. This 'translation' table will assist in attempts to equate the neurodevelopmental literature across species with either large or small differences in gestation and maturation, and also permit studies done in a variety of mammals to be applied to better understand human development. The comparative data indicate that humans, although conventionally considered an altricial species, are neurally advanced at birth relative to the other species studied. PMID- 11483297 TI - High-resolution two-dimensional spatial mapping of cat striate cortex using a 100 microelectrode array. AB - Much of our understanding of the visuotopic organization of striate cortex results from single-electrode penetrations and serial recording of receptive field properties. However, the quality of these maps is limited by imprecision in quantifying electrode position, combining data from multiple laminae, and eye drift during the measurement of the receptive field properties. We have addressed these concerns by using an array of 100 closely spaced microelectrodes to investigate the two-dimensional visuotopic organization of layer IV in cat striate cortex. This array allowed simultaneous measurement of the receptive field properties of multiple single units on a regularly spaced grid. We found the relationship between cortical and visual space to be locally non-conformal: the receptive field locations associated with a closely spaced line of electrodes appeared randomly scattered in visual space. To quantify the scatter, we fitted a linear transformation of electrode sites onto the associated receptive field locations. We found that the distribution of the difference between the predicted receptive field location and the measured location had standard deviations of 0.59 degrees and 0.45 degrees in the horizontal and the vertical axes, respectively. Although individual receptive field positions differed from the predicted locations in a non-conformal sense, the trend across multiple receptive fields followed the maps described elsewhere. We found, on average, that the 13 mm2 of cortex sampled by the array mapped onto a 5.8-degrees) region of visual space. From the scaling of this map and a combination of the statistics of the receptive field size (2.7+/-1.5 degrees) and scatter, we have explored the impact of electrode spacing on the completeness and redundancy in coverage of visual space sampled by an array. The simulation indicated an array with 1.2-mm spacing would completely sample the region of visual space addressed by the array. These results have implications for neuroprosthetic applications. Assuming phosphene organization resembles the visuotopic organization, remapping of visual space may be necessary to accommodate the scatter in phosphene locations. PMID- 11483298 TI - Proliferation and differentiation of progenitor cells in the cortex and the subventricular zone in the adult rat after focal cerebral ischemia. AB - Progenitor cells in the subventricular zone of the lateral ventricle and in the dentate gyrus of the hippocampus can proliferate throughout the life of the animal. To examine the proliferation and fate of progenitor cells in the subventricular zone and dentate gyrus after focal cerebral ischemia, we measured the temporal and spatial profiles of proliferation of cells and the phenotypic fate of proliferating cells in ischemic brain in a model of embolic middle cerebral artery occlusion in the adult rat. Proliferating cells were labeled by injection of bromodeoxyuridine (BrdU) in a pulse or a cumulative protocol. To determine the temporal profile of proliferating cells, ischemic rats were injected with BrdU every 4 h for 12 h on the day preceding death. Rats were killed 2-14 days after ischemia. We observed significant increases in numbers of proliferating cells in the ipsilateral cortex and subventricular zone 2-14 days with a peak at 7 days after ischemia compared with the control group. To maximize labeling of proliferating cells, a single daily injection of BrdU was administered over a 14-day period starting the day after ischemia. Rats were killed either 2 h or 28 days after the last injection of BrdU. A significant increase in numbers of BrdU immunoreactive cells in the subventricular zone was coincident with a significant increase in numbers of BrdU immunoreactive cells in the olfactory bulb 14 days after ischemia and numbers of BrdU immunoreactive cells did not significantly increase in the dentate gyrus. However, 28 days after the last labeling, the number of BrdU labeled cells decreased by 90% compared with number at 14 days. Clusters of BrdU labeled cells were present in the cortex distal to the infarction. Numerous cells immunostained for the polysialylated form of the neuronal cell adhesion molecule were detected in the ipsilateral subventricular zone. Only 6% of BrdU labeled cells exhibited glial fibrillary acidic protein immunoreactivity in the cortex and subcortex and no BrdU labeled cells expressed neuronal protein markers (neural nuclear protein and microtubule associated protein-2). From these data we suggest that focal cerebral ischemia induces transient and regional specific increases in cell proliferation in the ipsilateral hemisphere and that proliferating progenitor cells may exist in the adult cortex. PMID- 11483299 TI - Dynamic induction of the long pentraxin PTX3 in the CNS after limbic seizures: evidence for a protective role in seizure-induced neurodegeneration. AB - Pentraxin 3, a prototypic long pentraxin, is induced by proinflammatory signals in the brain. Inflammatory cytokines are rapidly induced in glia by epileptic activity. We show that pentraxin 3 immunoreactivity and mRNA are enhanced in the rat forebrain above undetectable control levels by limbic seizures with a dual pattern of induction. Within 6 h from seizure onset, pentraxin 3 immunoreactivity was increased in astrocytes. Eighteen to 48 h later, specific neuronal populations and leucocytes were strongly immunoreactive only in areas of neurodegeneration. This staining was abolished when neuronal cell loss, but not seizures, was prevented by blocking N-methyl-D-aspartate receptors. Pentraxin 3 /- mice had a more widespread seizure-related neuronal damage in the forebrain than their wild-type littermates although both groups had similar epileptic activity. Our results provide evidence that pentraxin 3 is synthesized in brain after seizures and may exert a protective role in seizure-induced neurodegeneration. PMID- 11483300 TI - Adenosine preferentially suppresses serotonin2A receptor-enhanced excitatory postsynaptic currents in layer V neurons of the rat medial prefrontal cortex. AB - Serotonin induces 'spontaneous' (non-electrically evoked) excitatory postsynaptic currents in layer V pyramidal neurons in the prefrontal cortex. This is likely due to a serotonin2A receptor-mediated focal release of glutamate onto apical dendrites. In addition, activation of the serotonin2A receptor selectively enhances late components of electrically evoked excitatory postsynaptic currents. In this study, using in vitro intracellular and whole-cell recording in rat brain slices, we examined the role of adenosine in modulating serotonin2A-enhanced 'spontaneous' and electrically evoked excitatory postsynaptic currents in layer V pyramidal neurons in the medial prefrontal cortex. Adenosine and N6 cyclopentyladenosine, an A1 adenosine agonist, markedly suppressed the serotonin2A-induced ('spontaneous') excitatory postsynaptic currents. However, adenosine had no effect on spontaneous miniature (tetrodotoxin-insensitive) postsynaptic potentials. Adenosine also blocked the late excitatory postsynaptic currents induced by the serotonin2A/2C agonist R(-)-2,5-dimethoxy-4 iodoamphetamine hydrochloride. Surprisingly, in contrast to other regions, adenosine had a relatively small effect on electrically evoked fast excitatory postsynaptic currents. These findings represent a novel demonstration of adenosine's ability to preferentially modulate serotonin2A-mediated synaptic events in the medial prefrontal cortex. As the serotonin2A receptor is closely linked with the effects of atypical antipsychotics and hallucinogens, further understanding of the modulators of this receptor such as adenosine may provide useful therapeutic applications. PMID- 11483301 TI - Activation of presynaptic group III metabotropic glutamate receptors depresses spontaneous inhibition in layer V of the rat entorhinal cortex. AB - Whole cell voltage clamp recording was used to investigate neurotransmitter release onto neurones in deep and superficial layers of rat entorhinal cortex in vitro. Activation of metabotropic glutamate receptors with the agonist (1S,3R,4S) 1-aminocyclopentane-1,2,4-tricarboxylic acid depressed spontaneous release of the inhibitory neurotransmitter GABA in layer V, but not in layer II. Depression of transmitter release did not persist in the presence of the sodium channel blocker tetrodotoxin. It seems likely that activation of presynaptic glutamate heteroreceptors inhibits action potential dependent release of neurotransmitter via a direct action at the presynaptic terminal. We confirmed that depression of inhibitory neurotransmission in layer V was mediated by group III metabotropic glutamate receptors using a specific group III antagonist, (RS)-cyclopropyl-4 phosphonophenylglycine. Application of the antagonist alone did not alter the frequency of spontaneous neurotransmitter release, suggesting that the metabotropic glutamate receptor is not tonically active. In layer V of the entorhinal cortex, activation of presynaptic metabotropic glutamate receptors enhances spontaneous glutamate release, and inhibits spontaneous release of GABA. These effects may combine to increase random action potential firing in this layer, thereby reducing its capacity for synchrony generation. Our results are consistent with an anticonvulsant action for group III metabotropic glutamate receptors in the entorhinal cortex. PMID- 11483302 TI - Nitric oxide is involved in ischemia-induced apoptosis in brain: a study in neuronal nitric oxide synthase null mice. AB - Nitric oxide can promote or inhibit apoptosis depending on the cell type and coexisting metabolic or experimental conditions. We examined the impact of nitric oxide on development of apoptosis 6, 24, and 72 h after permanent middle cerebral artery occlusion in mutant mice that lack the ability to generate nitric oxide from neuronal nitric oxide synthase. Adjacent coronal sections passing through the anterior commissure were stained with hematoxylin and eosin or terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). Immunoblotting was used to identify changes in the anti- and proapoptotic proteins Bcl-2 and Bax, respectively. Activation of caspases was assessed by appearance of actin cleavage products using a novel antiserum directed against 32 kDa actin fragment (fractin). In the neuronal nitric oxide synthase mutant mouse, infarct size and TUNEL positive apoptotic neurons were reduced compared to the wild-type controls. At 6 h, Bcl-2 levels in the ischemic hemisphere were increased in mutants but decreased in the wild-type strain. Bax levels did not change significantly. Caspase-mediated actin cleavage appeared in the ischemic hemisphere at this time point, and was significantly less in mutant brains at 72 h compared to the wild-type. The reduction in the number of TUNEL and fractin positive apoptotic cells appears far greater than anticipated based on the smaller lesion size in mutant mice.Hence, from these data we suggest that a deficiency in neuronal nitric oxide production slows the development of apoptotic cell death after ischemic injury and is associated with preserved Bcl-2 levels and delayed activation of effector caspases. PMID- 11483303 TI - Distribution of the high-affinity choline transporter in the central nervous system of the rat. AB - In cholinergic nerve terminals, Na(+)- and Cl(-)-dependent, hemicholinium-3 sensitive, high-affinity choline uptake is thought to be the rate-limiting step in acetylcholine synthesis. The high-affinity choline transporter cDNA responsible for the activity was recently cloned. Here we report production of a highly specific antibody to the high-affinity choline transporter and distribution of the protein in the CNS of the rat. The antibody stained almost all known cholinergic neurons and their terminal fields. High-affinity choline transporter-immunoreactive cell bodies were demonstrated in the olfactory tubercle, basal forebrain complex, striatum, mesopontine complex, medial habenula, cranial nerve motor nuclei, and ventral horn and intermediate zone of the spinal cord. Noticeably, high densities of high-affinity choline transporter positive axonal fibers and puncta were encountered in many brain regions such as cerebral cortex, hippocampus, amygdala, striatum, several thalamic nuclei, and brainstem. Transection of the hypoglossal nerve resulted in a loss of high affinity choline transporter immunoreactivity in neurons within the ipsilateral hypoglossal motor nucleus, which paralleled a loss of immunoreactivity to choline acetyltransferase. The antibody also stained brain sections from human and mouse, suggesting cross-reactivity. These results confirm that the high-affinity choline transporter is uniquely expressed in cholinergic neurons and is efficiently transported to axon terminals. The antibody will be useful to investigate possible changes in cholinergic cell bodies and axon terminals in human and rodents under various pathological conditions. PMID- 11483304 TI - The morphological phenotype of beta-amyloid plaques and associated neuritic changes in Alzheimer's disease. AB - We have utilised laser confocal microscopy to categorise beta-amyloid plaque types that are associated with preclinical and end-stage Alzheimer's disease and to define the neurochemistry of dystrophic neurites associated with various forms of plaques. Plaques with a spherical profile were defined as either diffuse, fibrillar or dense-cored using Thioflavin S staining or immunolabelling for beta amyloid. Confocal analysis demonstrated that fibrillar plaques had a central mass of beta-amyloid with compact spoke-like extensions leading to a confluent outer rim. Dense-cored plaques had a compacted central mass surrounded by an outer sphere of beta-amyloid. Diffuse plaques lacked a morphologically identifiable substructure, resembling a ball of homogeneous labelling. The relative proportion of diffuse, fibrillar and dense-cored plaques was 53, 22 and 25% in preclinical and 31, 49 and 20% in end-stage Alzheimer's disease cases, respectively. Plaque associated dystrophic neurites in preclinical cases were immunolabeled for neurofilament proteins whereas, in end-stage cases, these abnormal neurites were variably labelled for tau and/or neurofilaments. Double labelling demonstrated that the proportion of diffuse, fibrillar and dense-cored plaques that were neuritic was 12, 47 and 82% and 24, 82 and 76% in preclinical and end-stage cases, respectively. Most dystrophic neurites in Alzheimer's disease cases were labelled for either neurofilaments or tau, however, confocal analysis determined that 30% of neurofilament-labelled bulb-like or elongated neurites had a core of tau immunoreactivity. These results indicate that all morphologically defined beta-amyloid plaque variants were present in both early and late stages of Alzheimer's disease. However, progression to clinical dementia was associated with both a shift to a higher proportion of fibrillar plaques that induced local neuritic alterations and a transformation of cytoskeletal proteins within associated abnormal neuronal processes. There data indicate key pathological changes that may be subject to therapeutic intervention to slow the progression of Alzheimer's disease. PMID- 11483305 TI - Sodium currents in isolated rat CA1 pyramidal and dentate granule neurones in the post-status epilepticus model of epilepsy. AB - Status epilepticus (SE) was induced in the rat by long-lasting electrical stimulation of the hippocampus. After a latent period of 1 week, spontaneous seizures occurred which increased in frequency and severity in the following weeks, finally culminating after 3 months in a chronic epileptic state. In these animals we determined the properties of voltage-dependent sodium currents in acutely isolated CA1 pyramidal neurones and dentate granule cells using the whole cell voltage-clamp technique. The conductance of the fast transient sodium current was larger in SE rats (84+/-7 nS versus 56+/-6 nS) but related to a difference in cell size so that the neurones had a similar specific sodium conductance (control: 7.8+/-0.8 nS/pF, SE: 6.7+/-0.8 nS/pF). Current activation and inactivation were characterised by a Boltzmann function. After SE the voltage dependence of activation was shifted to more negative potentials (control: 45.1+/-1.4 mV, SE: -51.5+/-2.9 mV, P<0.05). In combination with a small shift in the voltage dependence of inactivation to more depolarised potentials (control: 68.8+/-2.3 mV, SE: -66.3+/-2.3 mV), it resulted in a window current that was much increased in the SE neurones (median: 64 pA in control, 217 pA in SE, P<0.05). The peak of this window current shifted to more hyperpolarised potentials (control: -44 mV, SE: -50 mV, P<0.05). No differences were found in the sodium currents analysed in dentate granule cells of control and SE animals. The changes observed in CA1 neurones after SE contribute to enhanced excitability in particular when membrane potential is near firing threshold. They can, at least partly, explain the lower threshold for epileptic activity in SE animals. The comparison of CA1 with DG neurones in the same rats demonstrates a differential response in the two cell types that participated in very similar seizure activity. PMID- 11483306 TI - Action potential threshold of hippocampal pyramidal cells in vivo is increased by recent spiking activity. AB - Understanding the mechanisms that influence the initiation of action potentials in single neurons is an important step in determining the way information is processed by neural networks. Therefore, we have investigated the properties of action potential thresholds for hippocampal neurons using in vivo intracellular recording methods in Sprague-Dawley rats. The use of in vivo recording has the advantage of the presence of naturally occurring spatio-temporal patterns of synaptic activity which lead to action potential initiation. We have found there is a large variability in the threshold voltage (5.7+/-1.7 mV; n=22) of individual action potentials. We have identified two separate factors that contribute to this variation in threshold: (1) fast rates of membrane potential change prior to the action potential are associated with more hyperpolarized thresholds (increased excitability) and (2) the occurrence of other action potentials in the 1 s prior to any given action potential is associated with more depolarized thresholds (decreased excitability). We suggest that prior action potentials cause sodium channel inactivation that recovers with approximately a 1 s time constant and thus depresses action potential threshold during this period. PMID- 11483307 TI - Post-insult activity is a major cause of delayed neuronal death in organotypic hippocampal slices exposed to glutamate. AB - We investigated the pathophysiological mechanisms of glutamate-induced delayed neuronal damage in rat hippocampal slice cultures [Stoppini et al. (1991) J. Neurosci. Methods 37, 173-182], with propidium iodide as a marker of cell death. Exposure of the cultures to growth medium containing 10 mM glutamate for 30 min resulted in a slowly developing degeneration of hippocampal principal cells, starting from the medial end of the CA1 region and reaching the dentate gyrus by 48 h. By 24 h, most pyramidal cells in CA1 were damaged. An acute phase of degeneration preceded the delayed damage at 2-6 h, affecting cells in a spatially diffuse manner. When tetrodotoxin (0.5 microM) was present during the glutamate insult, a marked protection (mean 57%, P<0.001) of the CA1 damage was observed. Rather strikingly, when tetrodotoxin was applied immediately following or even with a delay of 30 min after the insult, a similar amount of protection was achieved. In field recordings carried out after the insult, the glutamate-treated slices exhibited spontaneously occurring negative shifts with a duration of 1-10 s and an amplitude of up to 400 microV in the CA3 region, whereas the control slices were always quiescent. Taken together, the results suggest that post insult neuronal network activity, rather than the direct action of exogenous glutamate, is a major cause of delayed CA1 pyramidal cell death in the organotypic slices. These observations may have implications in the design of neuroprotective strategies for the treatment of brain traumas which are accompanied by delayed and/or distal neuronal damage. PMID- 11483308 TI - Differential changes of synaptic transmission in spiny neurons of rat neostriatum following transient forebrain ischemia. AB - Spiny neurons in neostriatum are vulnerable to cerebral ischemia. To reveal the mechanisms underlying the postischemic neuronal damage, the spontaneous activities, evoked postsynaptic potentials and membrane properties of spiny neurons in rat neostriatum were compared before and after transient forebrain ischemia using intracellular recording and staining techniques in vivo. In control animals the membrane properties of spiny neurons were about the same between the left and right neostriatum but the inhibitory synaptic transmission was stronger in the left striatum. After severe ischemia, the spontaneous firing and membrane potential fluctuation of spiny neurons dramatically reduced. The cortically evoked initial excitatory postsynaptic potentials were suppressed after ischemia indicated by the increase of stimulus threshold and the rise time of these components. The paired-pulse facilitation test indicated that such suppression might involve presynaptic mechanisms. The inhibitory postsynaptic potentials in spiny neurons were completely abolished after ischemia and never returned to the control levels. A late depolarizing postsynaptic potential that was elicited from approximately 5% of the control neurons by cortical stimulation could be evoked from approximately 30% of the neurons in the left striatum and approximately 50% in the right striatum after ischemia. The late depolarizing postsynaptic potential could not be induced after acute thalamic transection. The intrinsic excitability of spiny neurons was suppressed after ischemia evidenced by the significant increase of spike threshold and rheobase as well as the decrease of repetitive firing rate following ischemia. The membrane input resistance and time constant increased within 6 h following ischemia and the amplitude of fast afterhyperpolarization significantly increased after ischemia. These results indicate the depression of excitatory monosynaptic transmission, inhibitory synaptic transmission and excitability of spiny neurons after transient forebrain ischemia whereas the excitatory polysynaptic transmission in neostriatum was potentiated. The facilitation of excitatory polysynaptic transmission is stronger in the right neostriatum than in the left neostriatum after ischemia. The suppression of inhibitory component and the facilitation of excitatory polysynaptic transmission may contribute to the pathogenesis of neuronal injury in neostriatum after transient cerebral ischemia. PMID- 11483309 TI - Behavioural correlates of striatal glial fibrillary acidic protein in the 3 nitropropionic acid rat model: disturbed walking pattern and spatial orientation. AB - The 3-nitropropionic acid animal model is a model where excitotoxicity, mitochondrial dysfunction and oxidative stress, mechanisms common to various neurodegenerative diseases, are involved. The present study investigated whether behavioural alterations in this model were related to striatal damage. Wistar and Lewis rats were exposed to 3-nitropropionic acid and their behavioural performance (open field, walking pattern and Morris Water Maze task) was tested after the injections and after a recovery period of 3 weeks. No changes in activity were found in the open field test. Altered walking pattern was observed in the footprint analysis, although a different response was observed in the Wistar rats compared to the Lewis rats. Initially increased latency times were observed during visual discrimination learning in the Morris Water Maze task in 3 nitropropionic acid-treated Wistar rats compared to Wistar controls. During spatial discrimination learning (invisible platform) in the Morris Water Maze task the swimming velocity was decreased in both rat strains as a result of 3 nitropropionic acid treatment. Increased striatal glial fibrillary acidic protein concentration in Wistar rats correlated with several parameters of the footprint analysis and with the latency and distance in visual as well as spatial discrimination learning in the Morris Water Maze. It is concluded that measurement of walking pattern and spatial orientation performance are sensitive indicators to monitor behavioural changes in relation to striatal degeneration in the 3-nitropropionic acid animal model. In addition, Lewis rats are less sensitive towards 3-nitropropionic acid treatment than Wistar rats. PMID- 11483310 TI - Hippocampal and cortical sensory gating in rats: effects of quinpirole microinjections in nucleus accumbens core and shell. AB - Sensory processing disturbances, as measured in the P50/sensory gating paradigm, have been linked to aberrant auditory information processing and sensory overload in schizophrenic patients. In this paradigm, the response to the second of paired click stimuli is attenuated by an inhibitory effect of the first stimulus. Sensory gating has been observed in most healthy human subjects and normal laboratory rats. Because mesolimbic dopamine has been implicated in other filtering disturbances such as prepulse inhibition of the acoustic startle response and given the fact that amphetamine and apomorphine have been shown to disrupt gating, this study was performed to investigate the role of mesolimbic dopamine in sensory gating. The dopamine D2 receptor agonist quinpirole (10 microg/0.5 microl) was injected bilaterally in nucleus accumbens core and shell and effects on cortical and hippocampal sensory gating were investigated. Also, effects of the dopamine D2 receptor antagonist haloperidol (0.1 mg/kg, subcutaneously) as pretreatment were studied. First, quinpirole significantly reduced both the amplitude to the first click and gating as measured in the cortex and in the hippocampus. There was a tendency for the quinpirole effects on hippocampal gating to be more pronounced in rats injected in the shell. Secondly, haloperidol did not antagonize effects of quinpirole on hippocampal parameters, whereas haloperidol pretreatment fully antagonized quinpirole effects on cortical parameters. In conclusion, gating can be significantly reduced when a dopamine agonist is specifically targeted at mesolimbic dopamine D2 receptors. However, an important consideration is that the dopaminergic effects in the present study on gating are predominantly mediated by the effects on the amplitude to the first click. This has also been suggested for systemic amphetamine injections in rats and schizophrenic patients. This casts doubt on whether dopamine receptor activation affects the putative inhibitory process between the first and the second stimulus. PMID- 11483311 TI - Medium-voltage 5-9-Hz oscillations give rise to spike-and-wave discharges in a genetic model of absence epilepsy: in vivo dual extracellular recording of thalamic relay and reticular neurons. AB - In humans with absence epilepsy, spike-and-wave discharges develop in the thalamocortical system during quiet immobile wakefulness or drowsiness. The present study examined the initial stage of the spontaneous development of spike and-wave discharges in Genetic Absence Epilepsy Rats from Strasbourg. Bilateral electrocorticograms were recorded in epileptic and non-epileptic rats under freely moving and undrugged conditions and under neuroleptanalgesia. Short lasting episodes of medium-voltage 5-9-Hz (mean=6-Hz) oscillations usually emerged spontaneously from a desynchronized electrocorticogram and in bilateral synchrony in both rat strains. These oscillations were distinguishable from sleep spindles regarding their internal frequency, duration, morphology, and moment of occurrence. Spontaneous spike-and-wave discharges developed from such synchronized medium-voltage oscillations, the spike-and-wave complex occurring at the same frequency as the 5-9-Hz wave. Because the thalamus is thought to play a significant role in the generation of spike-and-wave discharges, dual extracellular recording and juxtacellular labelling of relay and reticular neurons were conducted to study the thalamic cellular mechanisms associated with the generation of spike-and-wave discharges. During medium-voltage 5-9-Hz oscillations, discharges of relay and reticular cells had identical patterns in epileptic and non-epileptic rats, consisting of occasional single action potentials and/or bursts (interburst frequency of up to 6-8 Hz) in relay cells, and of rhythmic bursts (up to 12-15 Hz) in reticular neurons, these discharging in the burst mode almost always before relay neurons. The discharge frequency of reticular bursts decelerated to 6 Hz by the beginning of the spike-and-wave discharges. During these, relay and reticular neurons usually fired in synchrony a single action potential or a high-frequency burst of two or three action potentials and a high-frequency burst, respectively, about 12 ms before the spike component of the spike-and-wave complexes. The frequency of these corresponded to the maximal frequency of the thalamocortical burst discharges associated with 5-9 Hz oscillations. The patterns of relay and reticular phasic cellular firings associated with spike-and-wave discharges had temporal characteristics similar to those associated with medium-voltage 5-9-Hz oscillations, suggesting that these normal and epileptic oscillations are underlain by similar thalamic cellular mechanisms. In conclusion, medium-voltage 5-9-Hz oscillations in the thalamocortical loop give rise to spike-and-wave discharges. Such oscillations are not themselves sufficient to initiate spike-and-wave discharges, meaning that genetic factors render thalamocortical networks prone to generate epileptic electrical activity, possibly by decreasing the excitability threshold in reticular cells. While these GABAergic neurons play a key role in the synchronization of glutamatergic relay neurons during seizures, relay cells may participate significantly in the regulation of the recurrence of the spike-and wave complex. Furthermore, it is very likely that synchronization of relay and reticular cellular discharges during absence seizures is generated in part by corticothalamic inputs. PMID- 11483312 TI - Inhibition of the substantia nigra suppresses absences and clonic seizures in audiogenic rats, but not tonic seizures: evidence for seizure specificity of the nigral control. AB - GABAergic inhibition of the substantia nigra pars reticulata has been shown to suppress seizures in most models of epilepsy involving forebrain networks, such as absences or clonic seizures. No such antiepileptic effects were observed, however, in genetically audiogenic rats exhibiting tonic seizures generated in the brainstem. This suggests a constitutive dysfunction of the nigral GABAergic neurotransmission in this strain of rat or a selective action of the nigral control on specific networks. In the present study, we first confirmed that bilateral injection of muscimol (700 pmol/side) in the substantia nigra had no effect in Wistar rats with audiogenic seizures (Wistar AS). [3H]Muscimol autoradiography suggested a 40% reduced density of GABA(A) receptors in the substantia nigra of Wistar AS, whereas no change was observed in the cortex and the superior colliculus (superficial and intermediate layers), as compared to control animals. In Wistar AS where 40 repetitions of audiogenic stimulations progressively induced generalised convulsive seizures with both tonic and clonic components, bilateral injection of muscimol (350 pmol/side) in the substantia nigra suppressed the clonic component but had no effect on tonic seizures. In hybrid rats issued from cross-breeding between Wistar AS and rats with spontaneous absence seizures, bilateral injection of muscimol (18 pmol/side) in the substantia nigra abolished cortical spike-and-wave discharges, but had no effect on tonic audiogenic seizures at doses up to 700 pmol/side. These results show that despite a decreased number of GABA(A) receptors in the substantia nigra, inhibition of this structure in Wistar AS still leads to inhibition of seizures involving forebrain structures. These results confirm that GABAergic inhibition of the substantia nigra has antiepileptic effects through the control of forebrain circuits. They suggest that this control mechanism has no inhibitory effect on circuits underlying audiogenic tonic seizures. PMID- 11483313 TI - Different motivational effects induced by the endogenous mu-opioid receptor ligands endomorphin-1 and -2 in the mouse. AB - The present study was designed to investigate the motivational effects of the newly discovered endogenous mu-opioid receptor ligands, endomorphin-1 and endomorphin-2, using the conditioned place preference paradigm in mice. The binding properties of these peptides were first examined using an opioid binding assay. In membranes obtained from the mouse whole brain, the binding of [3H][D Ala2, NMePhe4, Gly(ol)5]enkephalin (DAMGO; mu), but not of [3H][D-Phe2, D Phe5]enkephalin (DPDPE; delta) or [3H]U69593 (kappa) selectively and concentration-dependently competed with that of endomorphin-1 and endomorphin-2, indicating that both endomorphin-1 and endomorphin-2 are specific ligands for mu opioid receptors in the brain. Endomorphin-1 (1-30 nmol/mouse) given i.c.v. produced a dose-related place preference. This effect was abolished by pre treatment with the mu-opioid receptor antagonist beta-funaltrexamine but not the delta-opioid receptor antagonist naltrindole or the kappa-opioid receptor antagonist nor-binaltorphimine. In contrast, endomorphin-2 (5.6 nmol/mouse) produced place aversion. This aversive effect was inhibited by nor binaltorphimine as well as beta-funaltrexamine, but not by naltrindole. The place aversion produced by endomorphin-2 was also attenuated by pre-treatment with antiserum against the endogenous kappa-opioid receptor ligand dynorphin A (1-17). These findings indicate that endomorphin-1 may produce its rewarding effect via mu-opioid receptors. On the other hand, the aversive effect induced by endomorphin-2 may be associated with the stimulation of endomorphin-1-insensitive mu-opioid receptors and the activation of dynorphinergic systems in the mouse brain. PMID- 11483314 TI - Cerebellar synaptic protein expression in schizophrenia. AB - A cortico-subcortico-cerebellar neural circuit has been postulated to be important in the pathophysiology of schizophrenia. This study investigated whether there are synaptic changes in the cerebellum to accompany its putative involvement in the disorder. We measured the expression of three synaptic proteins (synaptophysin, complexin I and complexin II) in the cerebellar cortex of 16 subjects with schizophrenia and 16 controls using in situ hybridisation histochemistry and immunoautoradiography. Complexin I and II are expressed predominantly by inhibitory and excitatory neurones respectively. In schizophrenia, synaptophysin mRNA was decreased, as was complexin II and its mRNA. Complexin I mRNA and protein levels were unaltered. Expression of the mRNAs in the rat cerebellum was unaffected by 2 weeks administration of antipsychotic drugs (haloperidol, chlorpromazine, risperidone, olanzapine or clozapine). We conclude that there is synaptic pathology in the cerebellum in schizophrenia. By disrupting neural circuits, the alterations may contribute to the cerebellar dysfunction thought to occur in the disorder. PMID- 11483315 TI - Properties of solitary tract neurones responding to peripheral arterial chemoreceptors. AB - Despite the highly integrated pattern of response evoked by peripheral chemoreceptor stimulation, limited information exists regarding the neurones within the nucleus of the solitary tract that mediate this reflex. Using a working heart-brainstem preparation, we describe evoked synaptic response patterns, some intrinsic membrane properties, location, morphology and axonal projections of physiologically characterised 'chemoreceptive' neurones located in the solitary tract nucleus in the rat. From 172 whole cell recordings, 56 neurones were identified as chemoreceptive since they responded to aortic injections of low doses of sodium cyanide (2-5 microg). Chemoreceptive neurones had a mean resting membrane potential of -52+/-1 mV and input resistance was 297+/-15 M(Omega) (n=56). Synaptic responses evoked included excitatory synaptic potentials alone, excitatory-inhibitory post-synaptic potential complexes, inhibitory synaptic potentials alone and central respiratory modulated synaptic potentials. Synaptic response latency data were obtained by stimulating electrically the solitary tract: the mean excitatory synaptic latency was 5.2+/ 0.4 ms (range 2.5-8.0 ms; n=17). Chemoreceptive neurones showed a heterogeneity in their intrinsic membrane properties: neurones displayed either steady state, augmenting or adapting firing responses to depolarising current injection and, in some neurones, either delayed excitation or rebound activity following hyperpolarising pulses. Eleven chemoreceptive neurones were labelled and provided the first morphological data of these cells. Labelled somata were detected dorsomedial or medial to the solitary tract spanning the obex. Neurones typically had three to eight primary dendrites which often entered the solitary tract as well as extending across the ipsilateral region of the nucleus of the solitary tract. Axons were mostly unmyelinated with boutons of the en passant variety and often ramified within the solitary tract nucleus as well as coursed towards the ipsilateral ventral medulla. In summary, this study provides new data on the neurophysiological, anatomical and morphological properties of nucleus of the solitary tract neurones responding to arterial chemoreceptors in the rat. PMID- 11483316 TI - Effect of peripheral nerve injury on dorsal root ganglion neurons in the C57 BL/6J mouse: marked changes both in cell numbers and neuropeptide expression. AB - Several types of changes have been reported to occur in dorsal root ganglia following peripheral nerve injury, including loss of neurons and increases and decreases in peptide expression. However, with regard to loss of neurons, results have not been consistent, presumably due to different quantitative methodologies employed and species analyzed. So far, most studies have been conducted on rats; however, with the fast development of the transgenic techniques, the mouse has become a standard model animal in primary sensory research. Therefore we used stereological methods to determine the number of neurons, as well as the expression of galanin message-associated peptide, a marker for galanin-expressing neurons, neuropeptide Y, and calcitonin gene-related peptide in lumbar 5 dorsal root ganglia of both control C57 BL/6J mice and in mice subjected to a 'mid thigh' sciatic nerve transection (axotomy). In control animals the total number of lumbar 5 dorsal root ganglion neurons was about 12000. Seven days after axotomy, 24% of the dorsal root ganglion neurons were lost (P<0.001), and 54% were lost 28 days after axotomy (P<0.001). With regard to the percentage of peptide-expressing neurons, the results obtained showed that both galanin message associated peptide (from <1% to about 21%) and neuropeptide Y (from <1% to about 16%) are upregulated, whereas calcitonin gene-related peptide is downregulated (from about 41% to about 14%) following axotomy. Results obtained with retrograde labeling of the axotomized dorsal root ganglion neurons indicate that the neuropeptide regulations may be even more pronounced, if the analysis is confined to the axotomized dorsal root ganglion neurons rather than including the entire neuron population. We also applied conventional profile-based counting methods to compare with the stereological data and, although the results were comparable considering the trends of changes following axotomy, the actual percentage obtained with the two methods differed markedly, both for neuropeptide Y- and, especially, for galanin message-associated peptide-positive neurons. These present results demonstrate that marked species differences exist with regard to the effect of nerve injury on dorsal root ganglion neurons. Thus, whereas no neuron loss is seen in rat up to 4 weeks after a 'mid-thigh' transection [Tandrup et al. (2000) J. Comp. Neurol. 422, 172-180], the present results indicate a dramatic loss already after 1 week in mouse. It is suggested that the proximity in physical distance of the lesion to the cell body is a critical factor for the survival of the target-deprived neurons. Finally, stereological methodology seems warranted when assessing the total number of neurons as well as changes in peptide regulations after axotomy in mouse. PMID- 11483317 TI - Hyperexcitability in sensory neurons of rats selected for high versus low neuropathic pain phenotype. AB - Selection line rats congenitally high or low for autotomy in the neuroma model of neuropathic pain (HA and LA rats) were found to be correspondingly high and low in a second type of neuropathic pain, the Chung model, which employs an alternative phenotypic endpoint, tactile allodynia. It has been proposed that both phenotypes reflect ectopic hyperexcitability in axotomized primary sensory neurons. To test this hypothesis we made in vitro recordings from sensory neurons in the L4 and 5 dorsal root ganglia. Baseline excitability was similar in HA and LA rats, and axotomy caused an increase in both lines. However, in the one neuronal subclass previously linked to neuropathic pain in these models the increase was significantly greater in HA than LA rats, and only at the time when pain scores in the two lines were diverging. Heritable differences in electrical response to axotomy in a specific afferent cell type appear to be a fundamental determinant of neuropathic pain. PMID- 11483318 TI - Is it time for a new formalism to categorize normal tissue radiation injury? PMID- 11483319 TI - Late radiation effects to the rectum and bladder in gynecologic cancer patients: the comparison of LENT/SOMA and RTOG/EORTC late-effects scoring systems. AB - PURPOSE: To test the correlation of LENT/SOMA and RTOG/EORTC late-effect scales for rectum and bladder, 116 cases with gynecologic malignancies that were treated with radiotherapy were assessed with both scales. METHODS AND MATERIALS: All cases had been treated at least 6 months before the date of assessment with external beam radiotherapy (50--54 Gy to midline) and 1--2 fractions of HDR brachytherapy (2 x 8.5 Gy to point-A for 32 inoperable cases; 1 x 9.25 Gy to 5--9 mm from the ovoid surface for 84 postoperative cases). The patients were questioned with both scales, and the correlation between the two scales was analyzed by Spearman's rho (rank correlation) test. RESULTS: There were 64 cases with uterine cervix carcinoma and 52 cases with endometrium carcinoma, The overall (external + brachy) doses to ICRU points were 57.8 +/- 3.8 Gy for rectum and 59.3 +/- 4.9 Gy for bladder. The statistical analysis of LENT/SOMA and RTOG/EORTC scales revealed a very good correlation for rectum (r = 0.81; p < 0.01) and a good correlation for bladder (r = 0.72; p < 0.01). CONCLUSION: The LENT/SOMA system is a further step on the reporting of late radiation effects. Some modifications will improve its precision, and multicentric randomized studies are needed to test its validity. PMID- 11483320 TI - Incorporating biologic measurements (SF(2), CFE) into a tumor control probability model increases their prognostic significance: a study in cervical carcinoma treated with radiation therapy. AB - PURPOSE: To assess whether incorporation of measurements of surviving fraction at 2 Gy (SF(2)) and colony-forming efficiency (CFE) into a tumor control probability (tcp) model increases their prognostic significance. METHODS AND MATERIALS: Measurements of SF(2) and CFE were available from a study on carcinoma of the cervix treated with radiation alone. These measurements, as well as tumor volume, dose, and treatment time, were incorporated into a Poisson tcp model (tcp(alpha,rho)). Regression analysis was performed to assess the prognostic power of tcp(alpha,rho) vs. the use of either tcp models with biologic parameters fixed to best-fit estimates (but incorporating individual dose, volume, and treatment time) or the use of SF(2) and CFE measurements alone. RESULTS: In a univariate regression analysis of 44 patients, tcp(alpha,rho) was a better prognostic factor for both local control and survival (p < 0.001 and p = 0.049, respectively) than SF(2) alone (p = 0.009 for local control, p = 0.29 for survival) or CFE alone (p = 0.015 for local control, p = 0.38 for survival). In multivariate analysis, tcp(alpha,rho) emerged as the most important prognostic factor for local control (p < 0.001, relative risk of 2.81). After allowing for tcp(alpha,rho), CFE was still a significant independent prognostic factor for local control, whereas SF(2) was not. The sensitivities of tcp(alpha,rho) and SF(2) as predictive tests for local control were 87% and 65%, respectively. Specificities were 70% and 77%, respectively. CONCLUSIONS: A Poisson tcp model incorporating individual SF(2), CFE, dose, tumor volume, and treatment time was found to be the best independent prognostic factor for local control and survival in cervical carcinoma patients. PMID- 11483321 TI - High-dose-rate brachytherapy in the treatment of uterine cervix cancer. Analysis of dose effectiveness and late complications. AB - PURPOSE: This retrospective analysis aims to report results of patients with cervix cancer treated by external beam radiotherapy (EBR) and high-dose-rate (HDR) brachytherapy. METHODS AND MATERIALS: From September 1992 to December 1996, 138 patients with FIGO Stages II and III and mean age of 56 years were treated. Median EBR to the whole pelvis was 45 Gy in 25 fractions. Parametrial boost was performed in 93% of patients, with a median dose of 14.4 Gy. Brachytherapy with HDR was performed during EBR or following its completion with a dose of 24 Gy in four weekly fractions of 6 Gy to point A. Median overall treatment time was of 60 days. Patient age, tumor stage, and overall treatment time were variables analyzed for survival and local control. Cumulative biologic effective dose (BED) at rectal and bladder reference points were correlated with late complications in these organs and dose of EBR at parametrium was correlated with small bowel complications. RESULTS: Median follow-up time was 38 months. Overall survival, disease-free survival, and local control at 5 years was 53.7%, 52.7%, and 62%, respectively. By multivariate and univariate analysis, overall treatment time up to 50 days was the only statistically significant adverse variable for overall survival (p = 0.003) and actuarial local control (p = 0.008). The 5-year actuarial incidence of rectal, bladder, and small bowel late complications was 16%, 11%, and 14%, respectively. Patients treated with cumulative BED at rectum points above 110 Gy(3) and at bladder point above 125 Gy(3) had a higher but not statistically significant 5-year actuarial rate of complications at these organs (18% vs. 12%, p = 0.49 and 17% vs. 9%, p = 0.20, respectively). Patients who received parametrial doses larger than 59 Gy had a higher 5-year actuarial rate of complications in the small bowel; however, this was not statistically significant (19% vs. 10%, p = 0.260). CONCLUSION: This series suggests that 45 Gy to the whole pelvis combined with four fractions of 6 Gy to point A with HDR brachytherapy is an effective and safe fractionation schedule in the treatment of Stages II and III cervix cancer if realized up to 50 days. To decrease the small bowel complications, we decreased the superior border of the parametrial fields to the S2-S3 level and the total dose to 54 Gy. PMID- 11483322 TI - Analysis of recurrence of squamous cell carcinoma of the uterine cervix after definitive radiation therapy alone: patterns of recurrence, latent periods, and prognosis. AB - PURPOSE: A retrospective analysis was performed with emphasis on the patterns of recurrence, latent period, and prognosis in patients with cervical squamous cell carcinoma of the uterus treated with definitive radiation therapy alone. Late recurrence, which was observed more than 5 years after the initial radiation therapy, was finally focused on and discussed. MATERIALS AND METHODS: Between 1976 and 1994, 256 patients with squamous cell carcinoma of the uterine cervix without hematogenous metastasis were treated with definitive radiation therapy alone. The patients were staged as follows according to the FIGO classification: 26 in Stage I, 56 in Stage II, 124 in Stage III, 28 in Stage IVa, and 22 in Stage IVb. All the patients were treated with external beam irradiation and low-dose rate intracavitary brachytherapy. RESULTS: A total of 74 patients had recurrence. The recurrence appeared in 67 cases (90.5%) within 5 years. Metastasis to para aortic and/or supraclavicular nodes developed later than other types of recurrence. Among patients with lymphogenous metastasis, there were more 5-year survivors after recurrence than with other types of recurrence. Patients with early recurrence, within 2 years of the initial therapy, had a worse prognosis than those with recurrence more than 2 years after treatment. Seven patients (2.7%) in all developed late recurrence more than 5 years after the treatment. The first site of recurrence was an abdominal para-aortic or supraclavicular node in all patients, excluding one patient who developed intrapelvic lymph node metastasis. Six patients had pelvic node metastasis detected with lymphangiography at the initial treatment. Median survival after late recurrence was 16.0 months. Two of 7 patients survived more than 3 years after secondary radiation therapy, and the remainder died of recurrent disease. CONCLUSION: Patients with para-aortic and/or supraclavicular node metastasis that developed late after the initial treatment are more likely to survive due to secondary radiation therapy. Careful follow-up is emphasized for long-term survivors. PMID- 11483323 TI - Significant pelvic recurrence in high-risk pathologic stage I--IV endometrial carcinoma patients after adjuvant chemotherapy alone: implications for adjuvant radiation therapy. AB - OBJECTIVE: To evaluate the risk of pelvic recurrence (PVR) in high-risk pathologic Stage I--IV endometrial carcinoma patients after adjuvant chemotherapy alone. METHODS: Between 1992 and 1998, 43 high-risk endometrial cancer patients received adjuvant chemotherapy. All patients underwent primary surgery consisting of total abdominal hysterectomy and bilateral salpingo-oophorectomy. No patients received preoperative radiation therapy (RT). Regional lymph nodes and peritoneal cytology were sampled in 62.8% and 83.7% of cases, respectively. Most patients had Stage III--IV disease (83.7%) or unfavorable histology tumors (74.4%). None had evidence of extra-abdominal disease. All patients received 4-6 cycles of chemotherapy as the sole adjuvant therapy, consisting primarily of cisplatin and doxorubicin. Recurrent disease sites were divided into pelvic (vaginal, nonvaginal) and extrapelvic (para-aortic, upper abdomen, liver, and extra abdominal). Median follow-up was 27 months (range, 2--96 months). RESULTS: Twenty nine women (67.4%) relapsed. Seventeen (39.5%) recurred in the pelvis and 23 (55.5%) in extrapelvic sites. The 3-year actuarial PVR rate was 46.5%. The most significant factors correlated with PVR were cervical involvement (CI) (p = 0.01) and adnexal (p = 0.05) involvement. Of the 17 women who developed a PVR, 8 relapsed in the vagina, 3 in the nonvaginal pelvis, and 6 in both. The 3-year vaginal and nonvaginal PVR rates were 37.8% and 26%, respectively. The most significant factor correlated with vaginal PVR was CI (p = 0.0007). Deep myometrial invasion (p = 0.02) and lymph nodal involvement (p = 0.03) were both correlated with nonvaginal PVR. Nine of the 29 relapsed patients (31%) developed PVR as their only (6) or first site (3) of recurrence. Factors associated with a higher rate of PVR (as the first or only site) were CI and Stage I--II disease. CONCLUSIONS: PVR is common in high-risk pathologic Stage I-IV endometrial cancer patients after adjuvant chemotherapy alone. These results support the continued use of locoregional RT in patients undergoing adjuvant chemotherapy. Further studies are needed to test the addition of chemotherapy to locoregional RT. PMID- 11483324 TI - Surgery and postoperative radiation therapy in FIGO Stage IIIC endometrial carcinoma. AB - OBJECTIVE: To determine the outcome, pattern(s) of failure, and optimal treatment volume in Stage IIIC endometrial carcinoma patients treated with surgery and postoperative radiation therapy (RT). METHODS: Between 1983 and 1998, 30 Stage IIIC endometrial carcinoma patients were treated with primary surgery and postoperative RT at the University of Chicago. All underwent total abdominal hysterectomy, bilateral salpingo-oophorectomy, sampling of pelvic lymph nodes (PLN), and peritoneal cytology. All were noted to have PLN involvement. Para aortic lymph nodes (PALN) were sampled in 26 cases, and were positive in 14 cases (54%). Twenty women received whole-pelvic RT (WPRT) and 10 (WPRT), plus paraortic RT (extended-field RT, EFRT). One EFRT patient also underwent concomitant whole abdominal RT (WART). Adjuvant vaginal brachytherapy (VB) was delivered in 10, chemotherapy in 5, and hormonal therapy in 7 patients. RESULTS: At a median follow-up of 32 months, the actuarial 5-year disease-free and cause-specific survivals of the entire group were 33.9% and 55.8%, respectively. Overall, 16 women (53%) relapsed. Sites of failure included the pelvis (23%), abdomen (13%), PALN (13%), and distant (40%). Of the 7 pelvic failures, 4 were vaginal (3 vaginal only). Patients treated with VB had a trend to a lower vaginal recurrence rate (0/10 vs. 4/20, p = 0.12) than those not receiving VB. All 4 PALN failures were in women treated with WPRT (2 negative, 1 unsampled, and 1 positive PALN). None of the 10 EFRT patients (2 negative, 8 positive PALN) recurred in the PALN. No patient developed an isolated abdominal recurrence. Two patients developed significant RT sequelae: chronic diarrhea in 1 patient treated with WPRT and VB, and small bowel obstruction in 1 patient treated with EFRT. CONCLUSION: FIGO Stage IIIC disease comprises a small percentage of endometrial carcinoma patients but carries a poor prognosis. Our failure pattern suggests that the optimal adjuvant RT volume is EFRT, even in women with negative PALN sampling. VB should also be administered to improve local control. The low rate of abdominal recurrence does not support the routine use of WART in these women. Given the predominance of failure in distant sites, attention should be focused on the development of systemic chemotherapy protocols. PMID- 11483325 TI - Intensified hyperfractionated accelerated radiotherapy limits the additional benefit of simultaneous chemotherapy--results of a multicentric randomized German trial in advanced head-and-neck cancer. AB - PURPOSE: To demonstrate the efficacy of radiochemotherapy (RCT) as the first choice of treatment for advanced unresectable head-and-neck cancer. To prove an expected benefit of simultaneously given chemotherapy, a two-arm randomized study with hyperfractionated accelerated radiochemotherapy (HF-ACC-RCT) vs. hyperfractionated accelerated radiotherapy (HF-ACC-RT) was initiated. The primary endpoint was 1-year survival with local control (SLC). METHODS AND MATERIALS: Patients with Stage III and IV (UICC) unresectable oro- and hypopharyngeal carcinomas were randomized for HF-ACC-RCT with 2 cycles of 5-FU (600 mg/m(2)/day)/carboplatinum (70 mg/m(2)) on days 1--5 and 29--33 (arm A) or HF-ACC RT alone (arm B). In both arms, there was a second randomization for testing the effect of prophylactically given G-CSF (263 microg, days 15--19) on mucosal toxicity. Total RT dose in both arms was 69.9 Gy in 38 days, with a concomitant boost regimen (weeks 1--3: 1.8 Gy/day, weeks 4 and 5: b.i.d. RT with 1.8 Gy/1.5 Gy). Between July 1995 and May 1999, 263 patients were randomized (median age 56 years; 96% Stage IV tumors, 4% Stage III tumors). RESULTS: This analysis is based on 240 patients: 113 patients with RCT and 127 patients with RT, qualified for protocol and starting treatment. There were 178 oropharyngeal and 62 hypopharyngeal carcinomas. Treatment was tolerable in both arms, with a higher mucosal toxicity after RCT. Restaging showed comparable nonsignificant different CR + PR rates of 92.4% after RCT and 87.9% after RT (p = 0.29). After a median observed time of 22.3 months, l- and 2-year local-regional control (LRC) rates were 69% and 51% after RCT and 58% and 45% after RT (p = 0.14). There was a significantly better 1-year SLC after RCT (58%) compared with RT (44%, p = 0.05). Patients with oropharyngeal carcinomas showed significantly better SLC after RCT (60%) vs. RT (40%, p = 0.01); the smaller group of hypopharyngeal carcinomas had no statistical benefit of RCT (p = 0.84). For both tumor locations, prophylactically given G-CSF was a poor prognostic factor (Cox regression), and resulted in reduced LRC (log-rank test: +/- G-CSF, p = 0.0072). CONCLUSION: With accelerated radiotherapy, the efficiency of simultaneously given chemotherapy may be not as high as expected when compared to standard fractionated RT. Oropharyngeal carcinomas showed better LRC after HF-ACC-RCT vs. HF-ACC-RT; hypopharyngeal carcinomas did not. Prophylactic G-CSF resulted in an unexpected reduced local control and should be given in radiotherapy regimen only with strong hematologic indication. PMID- 11483326 TI - Treatment results and prognostic factors of advanced T3--4 laryngeal carcinoma: the University of California, San Francisco (UCSF) and Stanford University Hospital (SUH) experience. AB - PURPOSE: To review the UCSF-SUH experience in the treatment of advanced T3--4 laryngeal carcinoma and to evaluate the different factors affecting locoregional control and survival. METHODS AND MATERIALS: We reviewed the records of 223 patients treated for T3--4 squamous cell carcinoma of the larynx between October 1, 1957, and December 1, 1999. There were 187 men and 36 women, with a median age of 60 years (range, 28--85 years). The primary site was glottic in 122 and supraglottic in 101 patients. We retrospectively staged the patients according to the 1997 AJCC staging system. One hundred and twenty-seven patients had T3 lesions, and 96 had T4 lesions; 132 had N0, 29 had N1, 45 had N2, and 17 had N3 disease. The overall stage was III in 93 and IV in 130 patients. Seventy-nine patients had cartilage involvement, and 144 did not. Surgery was the primary treatment modality in 161 patients, of which 134 had postoperative radiotherapy (RT), 11 had preoperative RT, 7 had surgery followed by RT and chemotherapy (CT), and 9 had surgery alone. Forty-one patients had RT alone, and 21 had CT with RT. Locoregional control (LRC) and overall survival (OS) were estimated using the Kaplan--Meier method. Log-rank statistics were employed to identify significant prognostic factors for OS and LRC. RESULTS: The median follow-up was 41 months (range, 2--367 months) for all patients and 78 months (range, 6--332 months) for alive patients. The LRC rate was 69% at 5 years and 68% at 10 years. Eighty-four patients relapsed, of which 53 were locoregional failures. Significant prognostic factors for LRC on univariate analysis were primary site, N stage, overall stage, the lowest hemoglobin (Hgb) level during RT, and treatment modality. Favorable prognostic factors for LRC on multivariate analysis were lower N stage and primary surgery. The overall survival rate was 48% at 5 years and 34% at 10 years. Significant prognostic factors for OS on univariate analysis were: primary site, age, overall stage, T stage, N stage, lowest Hgb level during RT, and treatment modality. Favorable prognostic factors for OS on multivariate analysis were lower N stage and higher Hgb level during RT. CONCLUSION: Lower N-stage was a favorable prognostic factor for LRC and OS. Hgb levels > or = 12.5 g/dL during RT was a favorable prognostic factor for OS. Surgery was a favorable prognostic factor for LRC but did not impact on OS. Correcting the Hbg level before and during treatment should be investigated in future clinical trials as a way of improving therapeutic outcome in patients with advanced laryngeal carcinomas. PMID- 11483327 TI - Is the 1997 AJCC staging system for nasopharyngeal carcinoma prognostically useful for Chinese patient populations? AB - PURPOSE: The 5th edition of the American Joint Committee on Cancer (AJCC) staging manual defines new rules for classifying nasopharyngeal carcinoma (NPC). The study was conducted to assess its effectiveness in predicting the prognosis for Chinese patient populations. METHODS AND MATERIALS: Between June 1993 and June 1994, 621 consecutively admitted patients with nondisseminated NPC were treated with definitive-intent radiation therapy alone. All had computed tomography of the nasopharynx, skull base, and the upper neck. A computer database containing all information for staging was formed on presentation. The extent of disease of each patient was restaged according to the 1997 AJCC system. RESULTS: Of the 621 patients, The 5-year overall survival (OS) rate was 60%. The 1997 AJCC system creates subgroups (Stages I to IV) that are assigned to 38 (6.1%), 270 (43.5%), 157 (25.3%), and 156 (25.1%) patients, respectively. The incidence of parapharyngeal extension was 74.1% (460/621). Of these patients (460) with parapharyngeal extension, 310 (67.4%) patients were classified as T2b disease. The 1997 AJCC system showed highly significant differences between the overall stages for both OS and relapse-free survival (RFS). The 1997 AJCC T classifications showed significant correlation with local failure, and N classification was accurate in predicting FDM. Multivariate analysis showed that paraoropharyngeal involvement was an independently significant prognostic factor for OS, freedom from local recurrence (FLR), and freedom form distant metastasis (FDM). CONCLUSION: The 1997 AJCC staging system for NPC is prognostically useful for Chinese patient populations. We proposed that subdivision of parapharyngeal extension should be included in future revisions of the staging system. PMID- 11483328 TI - The American Brachytherapy Society recommendations for high-dose-rate brachytherapy for head-and-neck carcinoma. AB - PURPOSE: To develop recommendations for use of high-dose-rate (HDR) brachytherapy in patients with head-and-neck cancer. METHODS: A panel consisting of members of the American Brachytherapy Society (ABS) performed a literature review, added information based upon their clinical experience, and formulated recommendations for head-and-neck HDR brachytherapy. RESULTS: The ABS recommends the use of brachytherapy as a component of the treatment of head-and-neck tumors. However, the ABS recognizes that some radiation oncologists are reluctant to employ brachytherapy in the head-and-neck region because of the complexity of the postoperative management and concerns about radiation safety. In this regard, HDR eliminates unwanted radiation exposure and thereby permits unrestricted delivery of clinical care to these brachytherapy patients. The ABS made specific recommendations for previously untreated and recurrent head-and-neck cancer patients on patient selection criteria, implant techniques, target volume definition, and HDR treatment parameters (such as time, dose, and fractionation schedules). Suggestions were provided for treatment with HDR alone and in combination with external beam radiation therapy. It should be recognized that only limited experiences exist with HDR brachytherapy in patients with head-and neck cancers. Therefore, some of these suggested doses have not been extensively tested in clinical practice. Hence, these guidelines will be updated as significant new outcome data are available. Any clinician following these guidelines is expected to use clinical judgment to determine an individual patient's treatment. CONCLUSIONS: Little has been published in the clinical literature on HDR brachytherapy in head-and-neck cancer. Based upon the available information and the clinical experience of the panel members, general and site specific recommendations were offered. Areas for further investigations were identified. PMID- 11483329 TI - Quality of life assessment in advanced non-small-cell lung cancer patients undergoing an accelerated radiotherapy regimen: report of ECOG study 4593. Eastern Cooperative Oncology Group. AB - PURPOSE: To prospectively evaluate the quality of life (QOL) before, at completion, and after therapy for patients receiving an accelerated fractionation schedule of radiotherapy for advanced, unresectable non-small-cell lung cancer in a Phase II multi-institutional trial. METHODS AND MATERIALS: The Functional Assessment of Cancer Therapy-Lung (FACT-L) patient questionnaire was used to score the QOL in patients enrolled in the Eastern Cooperative Oncology Group Phase II trial (ECOG 4593) of hyperfractionated accelerated radiotherapy in non small-cell lung cancer. Radiotherapy (total dose 57.6 Gy in 36 fractions) was delivered during 15 days, with three radiation fractions given each treatment day. The protocol was activated in 1993, and 30 patients had accrued by November 1995. The FACT-L questionnaire was administered at study entry (baseline), on the last day of radiotherapy (assessment 2), and 4 weeks after therapy (assessment 3). The FACT-L includes scores for physical, functional, emotional, and social well-being (33 items), and a subscale of lung cancer symptoms (10 additional items). The summation of the physical, functional, and lung cancer symptom subscales (21 items) constitutes the Trial Outcome Index (TOI), considered the most clinically relevant outcome measure in lung cancer treatment trials. RESULTS: The FACT-L completion rates at the designated study time points were as follows: baseline, 30 of 30 (100%); assessment 2, 29 (97%) of 30; and assessment 3, 24 (80%) of 30. At treatment completion, statistically significant declines in QOL scores were noted, compared with baseline for physical and functional well being. Emotional well-being scores improved at both assessment 2 and assessment 3. The physical and functional scores returned approximately to baseline values at assessment 3. The change in TOI score was evaluated as a function of the clinical response to treatment, toxicity grade, and survival; no clear association was noted. A trend for the largest decrease in QOL was noted for patient groups with shorter survival times. The mean change in the TOI score from baseline to assessment 3 was -8.96 for patients surviving < 52 weeks vs. -0.95 for those surviving > 52 weeks. CONCLUSIONS: The FACT-L questionnaire can be successfully administered to non-small-cell lung cancer patients enrolled in a prospective Phase II trial of accelerated radiation fractionation. The decrement in physical and functional QOL during treatment returned to baseline level at 4 weeks after treatment. Emotional well-being improved at all time points. A trend was noted for shorter survival times in patients with the largest negative change in TOI score. These data suggest that the clinical use of hyperfractionated accelerated radiotherapy did not cause a significant, long-term decrease in the QOL of the treated patients, and that it is feasible to perform a QOL study of patients enrolled in such a trial. PMID- 11483330 TI - Temporary PSA rises and repeat prostate biopsies after brachytherapy. AB - PURPOSE: The long-standing confusion regarding the clinical relevance of postimplant biopsies is complicated by the common occurrence of temporary PSA rises between 1 and 2 years after brachytherapy. We report here 4 patients with temporary, self-limited PSA rises and postimplant biopsies, for whom radical prostatectomy was strongly advised but for whom surgery would probably have been the wrong choice. MATERIALS AND METHODS: Transperineal I-125 or Pd-103 implants were performed as previously described. After implantation, patients were followed routinely, with repeat PSA and physical examination at approximately every 4 to 6 months. Timing of postimplant PSAs was at the discretion of the patient and his doctors. Postimplant biopsies were performed in all cases out of concern for a persistently elevated serum PSA. Sections of fixed and embedded tissue were stained with standard hematoxylin and eosin. RESULTS: All 4 patients presented here were advised to have a salvage prostatectomy based primarily on their PSA changes. However, all of the patients have subsequently had a dramatic PSA fall, consistent with long-term cancer control, despite the fact that 3 of the 4 had histologic evidence of persistent cancer on repeat prostate biopsy. CONCLUSIONS: It is crucial that clinicians be aware of the potential for the doubly confusing situation of temporary PSA rises and apparently positive rebiopsies and the pressure it puts on both patients and their physicians to go ahead with inappropriate salvage therapy. PMID- 11483331 TI - Definitions of biochemical failure in prostate cancer following radiation therapy. AB - PURPOSE: The American Society for Therapeutic Radiology and Oncology (ASTRO) published a consensus panel definition of biochemical failure following radiation therapy for prostate cancer. In this paper, we develop a series of alternative definitions of biochemical failure. Using data from 688 patients, we evaluated the sensitivity and specificity of the various definitions, with respect to a defined "clinically meaningful" outcome. METHODS AND MATERIALS: The ASTRO definition of biochemical failure requires 3 consecutive rises in prostate specific antigen (PSA). We considered several modifications to the standard definition: to require PSA rises of a certain magnitude, to consider 2 instead of 3 rises, to require the final PSA value to be greater than a fixed cutoff level, and to define biochemical failure based on the slope of PSA over 1, 1.5, or 2 years. A clinically meaningful failure is defined as local recurrence, distant metastases, initiation of unplanned hormonal therapy, unplanned radical prostatectomy, or a PSA > 25 later than 6 months after radiation. RESULTS: Requiring the final PSA in a series of consecutive rises to be larger than 1.5 ng/mL increased the specificity of biochemical failure. For a fixed specificity, defining biochemical failure based on 2 consecutive rises, or the slope over the last year, could increase the sensitivity by up to approximately 20%, compared to the ASTRO definition. Using a rule based on the slope over the previous year or 2 rises leads to a slightly earlier detection of biochemical failure than does the ASTRO definition. Even with the best rule, only approximately 20% of true failures are biochemically detected more than 1 year before the clinically meaningful event time. CONCLUSION: There is potential for improvement in the ASTRO consensus definition of biochemical failure. Further research is needed, in studies with long follow-up times, to evaluate the relationship between various definitions of biochemical failure and true clinical outcome. PMID- 11483332 TI - Serum PSA evaluations during salvage radiotherapy for post-prostatectomy biochemical failures as prognosticators for treatment outcomes. AB - INTRODUCTION: Serum prostate specific antigen (PSA) levels have proved to be sensitive markers for the diagnosis of prostate cancer. In addition, PSA levels are useful for detecting and monitoring prostate cancer progression after radiotherapy. Serum PSA evaluations during radiotherapy, however, have not been well documented. In this study, we investigate the prognostic value of PSA evaluations during salvage radiotherapy for prostatectomy failures. METHODS: Forty-one patients with biochemical failures after prostatectomy treated with salvage radiotherapy consented to have their serum PSA levels evaluated at 30 Gy and 45 Gy of irradiation. All 41 patients had negative metastatic workup and pathologically uninvolved pelvic lymph nodes at the time of referral for salvage radiotherapy. Radiation therapy was delivered with 10--25 MV photons, with doses of 59.4--66.6 Gy. No patients received hormonal ablation therapy before irradiation. RESULTS: The mean follow-up for all patients was 30.9 months. At last follow-up, 28/41 patients (68.3%) were free from biochemical failure, with 20 of 41 patients (48.8%) expressing undetectable PSA levels. Serum PSA evaluations at 30 Gy did not significantly predict for either biochemical (p = 0.0917) or clinical (p = 0.106) disease-free outcome. However, serum PSA evaluations at 45 Gy significantly predicted for both biochemical (p = 0.0043) and clinical (p = 0.0244) disease-free outcomes, with PSA elevations at 45 Gy significantly associated with poor outcomes. On univariate analysis of prognosticators for biochemical failures, the following were significant: an elevation in serum PSA levels at 45 Gy, detectable serum PSA immediately after prostatectomy, Gleason score 7--10, and serum PSA level >1 ng/ml before salvage radiotherapy. CONCLUSION: Evaluation of serum PSA level at 45 Gy of salvage radiotherapy for biochemical relapses after prostatectomy may serve as a significant prognosticator for both biochemical and clinical disease-free outcomes. PMID- 11483333 TI - Improvement in dose escalation using the process of adaptive radiotherapy combined with three-dimensional conformal or intensity-modulated beams for prostate cancer. AB - PURPOSE: Advances in technology allow the creation of complex treatment plans with tightly conforming doses. However, variations in positioning of the organ/patient with respect to treatment beams necessitate the use of an appreciable margin, potentially limiting dose escalation in many patients. To (1) reduce this margin and (2) test the hypothesis that the achievable level of dose escalation is patient dependent, a patient-specific, confidence-limited planning target volume (cl-PTV) was constructed using an adaptive radiotherapy (ART) process for prostate cancer treatment developed in-house. The potential dose escalation achievable with this ART process is quantified for both conformal radiotherapy (CRT) delivery and intensity-modulated radiotherapy (IMRT) delivery. MATERIAL AND METHODS: Patients with organ confined prostate cancer were entered prospectively into an ART process developed in-house. This ART process has been designed to improve accuracy and precision of dose delivery, consequently enhancing dose escalation. In this process, a cl-PTV is constructed for each patient in the second week of treatment based upon on-line portal and CT images acquired during the first week of treatment. The treatment prescription dose, defined as the minimum dose to the cl-PTV, is selected based on predefined dose volume constraints for rectum/bladder and derived from the pretreatment planning CT image. In addition, the treatment modality (CRT or IMRT) is determined based on the level of dose escalation achievable and the risk of inaccurate targeting. The potential for both dose escalation and the application of IMRT was evaluated by comparing the prescription doses delivered using the ART process, with the cl PTV, to those in the traditional treatment process, with a conventional generic PTV. In addition, the distributions of potential geometric target underdosing and normal tissue overdosing were also calculated to evaluate the quality of the conventional treatment plans. RESULTS: One hundred and fifty patients have been treated with the ART process. When compared to the treatment dose delivered with the conventional treatment process (generic PTV), an average 5% (2.5--10%) more dose could be delivered using the ART process with CRT, and 7.5% (2.5--15%) more dose could be delivered with IMRT. Of the 150 patients, 70% were treated to a minimum cl-PTV dose > or = 77.4 Gy (81.3 Gy ICRU isocenter dose). Dosimetric analysis revealed that 81 Gy to the cl-PTV (or 86.7 Gy ICRU) could be prescribed to at least 50% of patients if IMRT was applied using the ART process. In contrast, IMRT did not yield an obvious dose escalation gain if patients were treated using the generic PTV. Our results also demonstrate that the cl-PTV is significantly smaller than the conventional generic PTV for most patients, with a mean volume reduction of 24% (range, 5--43%). CONCLUSION: These results support our hypothesis that the achievable level of dose escalation using ART is patient dependent. By using the ART process to develop a cl-PTV, one can (1) optimize the dose level, (2) increase the applicability of IMRT, and (3) improve the quality of dose delivery. The ART process provides the foundation to identify a suitable option (CRT or IMRT) for the delivery of a safe treatment and dose escalation. It is now our standard of practice for prostate cancer treatment. PMID- 11483334 TI - Potency after permanent prostate brachytherapy for localized prostate cancer. AB - PURPOSE: The evaluation of potency preservation after treatment of localized prostate cancer with transperineal permanent prostate brachytherapy (PPB) and the efficacy of sildenafil were studied. METHODS AND MATERIALS: This study comprised 482 patients who were able to maintain an erection suitable for intercourse before treatment from a cohort of 1166 patients with clinically localized prostate cancer treated with PPB. All patients have been followed prospectively, and actuarial analysis was performed to assess potency preservation over time. Patients treated with sildenafil were evaluated as to its efficacy. RESULTS: The median follow-up of this cohort was 34 months (6--92), with a median age of 68 years (47--80). Potency was preserved in 311 of the 482 patients, with a 5-year actuarial potency rate of 52.7%. The 5-year actuarial potency rate for patients treated with PPB as monotherapy was 76%, and, for those treated with combination external beam radiotherapy (EBT) + PPB, 56% (p = 0.08). Patients treated with neoadjuvant androgen deprivation (NAAD) + PPB had a 5-year potency rate of 52%, whereas those with combination EBT + PPB + NAAD had a potency rate of 29% (p = 0.13). Cox regression analysis identified that pretreatment use of NAAD and patient age predicted for impotence (p = 0.0001 and 0.04, respectively). Of 84 patients treated with sildenafil, 52 had a successful outcome (62%). The response to sildenafil was significantly better in those patients not treated with NAAD (p = 0.04). CONCLUSIONS: The actuarial potency rates at 5 years for patients treated with PPB are lower than generally acknowledged, except for those patients treated with PPB as monotherapy. Patients who received sildenafil exhibited improved potency in a majority of cases. PMID- 11483335 TI - Phase III radiation therapy oncology group (RTOG) trial 86-10 of androgen deprivation adjuvant to definitive radiotherapy in locally advanced carcinoma of the prostate. AB - PURPOSE: To test the hypothesis that androgen ablation before and during radiotherapy for locally advanced carcinoma of the prostate may, by reducing tumor bulk and enhancing tumor cell kill, improve locoregional control and ultimately survival. METHODS AND MATERIALS: The study was conducted from 1987 to 1991. Eligible patients were those with bulky tumors (T2--T4) with or without pelvic lymph node involvement and without evidence of distant metastases. They were randomized to receive goserelin, 3.6 mg every 4 weeks; and flutamide, 250 mg t.i.d. for 2 months before radiation therapy and during radiation therapy (Arm I), or radiation therapy alone (Arm II). Of 471 randomized patients, 456 were evaluable: 226 on Arm I and 230 on Arm II. RESULTS: As of November 1999, the median follow-up has reached 6.7 years for all patients and 8.6 years for alive patients. At 8 years, androgen ablation has been associated with an improvement in local control (42% vs. 30%, p = 0.016), reduction in the incidence of distant metastases (34% vs. 45%, p = 0.04), disease-free survival (33% vs. 21%, p = 0.004), biochemical disease-free survival = PSA <1.5 (24% vs. 10%, p < 0.0001), and cause-specific mortality (23% vs. 31%, p = 0.05). However, subset analysis indicates that the beneficial effect of short-term androgen ablation appears preferentially in patients with Gleason score 2--6. In that population, there is a highly significant improvement in all endpoints, including survival (70% vs. 52%, p = 0.015). In patients with Gleason 7--10 tumors, the regimen has not resulted in a significant enhancement in either locoregional control or survival. CONCLUSION: In patients with Gleason score 2--6 carcinoma of the prostate, a short course of androgen ablation administered before and during radiotherapy has been associated with a highly significant improvement in local control, reduction in disease progression, and overall survival. PMID- 11483336 TI - Five-year biochemical outcome after prostate brachytherapy for hormone-naive men < or = 62 years of age. AB - PURPOSE: To evaluate 5-year biochemical disease-free outcome for hormone naive men 62 years of age or less who underwent transperineal ultrasound-guided permanent prostate brachytherapy. METHODS AND MATERIALS: 76 patients underwent transperineal ultrasound guided prostate brachytherapy using either (103)Pd or (125)I for clinical T1b--T2b NxM0 (1997 AJCC) adenocarcinoma of the prostate gland from April 1995 to October 1999. No patient was lost to follow-up, and no patient underwent pathologic lymph-node staging. 47 patients were implanted with either (103)Pd or (125)I monotherapy, and 29 patients received moderate-dose external-beam radiation therapy followed by a prostate brachytherapy boost. No patient received hormonal manipulation. The median patient age was 58 years (range, 48--62 years). The median follow-up was 37 months (range, 14--70 months). Follow-up was calculated from the day of implantation. Biochemical disease-free survival was defined by the American Society of Therapeutic Radiation and Oncology (ASTRO) consensus definition. RESULTS: The actuarial 5-year biochemical disease-free survival rate was 98.7%. For patients with low-, intermediate-, and high-risk disease, 97.7%, 100%, and 100%, respectively, were free of biochemical failure. The median posttreatment prostate-specific antigen (PSA) for the entire group was 0.2 ng/mL. When stratified by risk group, the median posttreatment PSA was 0.2, 0.15, and 0.1 for patients with low-, intermediate-, and high-risk disease, respectively. CONCLUSION: With a median follow-up of 37 months, hormone naive patients < or = 62 years of age have a high probability of 5-year biochemical disease-free survival following permanent prostate brachytherapy with an apparent plateau on the PSA curve. PMID- 11483337 TI - Stage I and II MALT lymphoma: results of treatment with radiotherapy. AB - PURPOSE: Mucosa-associated lymphoid tissue (MALT) lymphoma is a distinct disease with specific clinical and pathologic features that may affect diverse organs. We analyzed our recent experience with Stage I/II MALT lymphoma presenting in the stomach and other organs to assess the outcome following involved field radiation therapy (RT). PATIENTS AND METHODS: Seventy patients with Stage IE (62) and IIE (8) disease were treated between 1989 and 1998. Patients with transformed MALT were excluded. The median age was 62 years (range, 24--83 years), M:F ratio 1:2.2. Presenting sites included stomach, 15; orbital adnexa, 19; salivary glands, 15; thyroid, 8; lung, 5; upper airways, 3 (nasopharynx, 2; larynx, 1); urinary bladder, 3; breast, 1; and rectum, 1. Staging included site-specific imaging, CT abdomen in 66 patients (94%) and bone marrow biopsy in 54 (77%). Sixty-two patients received radiation therapy: 52 received RT alone, 7 received chemotherapy and RT, and 3 received antibiotics followed by RT. Median RT dose was 30 Gy (range, 17.5--35 Gy). Most frequently used RT prescriptions were 25 Gy (26 patients-18 orbit, 6 stomach, and 2 salivary glands), 30 Gy (23 patients), and 35 Gy (8 patients). Five patients had complete surgical excision of lymphoma and no other treatment (stomach 1, salivary 2, lung 2), whereas 2 patients with gastric lymphoma received antibiotics only. One patient refused treatment and was excluded from the analysis of treatment outcome, leaving 69 patients with a median follow-up of 4.2 years (range, 0.3-11.4 years). RESULTS: A complete response was achieved in 66/69 patients, and 3 patients had partial response (2 lung, 1 orbit). The 5-year disease-free survival (DFS) was 76%, and the overall survival was 96%. No relapses were observed in patients with stomach and thyroid lymphoma. The 5-year DFS for these patients was 93%, in contrast to 69% for patients presenting in other sites (p = 0.006). Among the 5 patients treated with surgery only, 2 relapsed locally (lung, and minor salivary gland). Among 62 patients who received RT, 8 relapsed (2 salivary, 3 orbit, 1 nasopharynx, 1 larynx, 1 breast). Three patients relapsed in the nonirradiated contralateral paired organ, 4 in distant sites, and 1 in both local and distant sites. The overall local control rate with radiation was 97% (60/62 patients). CONCLUSION: Localized MALT lymphomas have excellent prognosis following moderate-dose RT. Gastric and thyroid MALT lymphomas have better early outcome, as compared to the other sites where distant failure is more common. Relapses were observed in nonirradiated paired organs or distant sites. Further follow-up is required to assess the impact of failure on survival. PMID- 11483338 TI - Stereotactic radiosurgery and fractionated stereotactic radiotherapy for the treatment of acoustic schwannomas: comparative observations of 125 patients treated at one institution. AB - BACKGROUND: Stereotactic radiosurgery (SRS) and, more recently, fractionated stereotactic radiotherapy (SRT) have been recognized as noninvasive alternatives to surgery for the treatment of acoustic schwannomas. We review our experience of acoustic tumor treatments at one institution using a gamma knife for SRS and the first commercial world installation of a dedicated linac for SRT. METHODS: Patients were treated with SRS on the gamma knife or SRT on the linac from October 1994 through August 2000. Gamma knife technique involved a fixed-frame multiple shot/high conformality single treatment, whereas linac technique involved daily conventional fraction treatments involving a relocatable frame, fewer isocenters, and high conformality established by noncoplanar arc beam shaping and differential beam weighting. RESULTS: Sixty-nine patients were treated on the gamma knife, and 56 patients were treated on the linac, with 1 NF 2 patient common to both units. Three patients were lost to follow-up, and in the remaining 122 patients, mean follow-up was 119 +/- 67 weeks for SRS patients and 115 +/- 96 weeks for SRT patients. Tumor control rates were high (> or =97%) for sporadic tumors in both groups but lower for NF-2 tumors in the SRT group. Cranial nerve morbidities were comparably low in both groups, with the exception of functional hearing preservation, which was 2.5-fold higher in patients who received conventional fraction SRT. CONCLUSION: SRS and SRT represent comparable noninvasive treatments for acoustic schwannomas in both sporadic and NF-2 patient groups. At 1-year follow-up, a significantly higher rate of serviceable hearing preservation was achieved in SRT sporadic tumor patients and may therefore be preferable to alternatives including surgery, SRS, or possibly observation in patients with serviceable hearing. PMID- 11483339 TI - Fractionated stereotactically guided radiotherapy and radiosurgery in the treatment of functional and nonfunctional adenomas of the pituitary gland. AB - PURPOSE: We evaluated survival rates and side effects after fractionated stereotactically guided radiotherapy (SCRT) and radiosurgery in patients with pituitary adenoma. METHODS AND MATERIALS: Between 1989 and 1998, 68 patients were treated with FSRT (n = 63) or radiosurgery (n = 5) for pituitary adenomas. Twenty six had functional and 42 had nonfunctional adenomas. Follow-up included CT/MRI, endocrinologic, and ophthalmologic examinations. Mean follow-up was 38.7 months. Seven patients received radiotherapy as primary treatment and 39 patients received it postoperatively for residual disease. Twenty-two patients were treated for recurrent disease after surgery. Mean total dose was 52.2 Gy for SCRT, and 15 Gy for radiosurgery. RESULTS: Overall local tumor control was 93% (60/65 patients). Forty-three patients had stable disease based on CT/MRI, while 15 had a reduction of tumor volume. After FSRT, 26% with a functional adenoma had a complete remission and 19% had a reduction of hormonal overproduction after 34 months' mean. Two patients with STH-secreting adenomas had an endocrinologic recurrence, one with an ACTH-secreting adenoma radiologic recurrence, within 54 months. Reduction of visual acuity was seen in 4 patients and partial hypopituitarism in 3 patients. None of the patients developed brain radionecrosis or radiation-induced gliomas. CONCLUSION: Stereotactically guided radiotherapy is effective and safe in the treatment of pituitary adenomas to improve local control and reduce hormonal overproduction. PMID- 11483340 TI - Thyroid dysfunction as a late effect in childhood medulloblastoma: a comparison of hyperfractionated versus conventionally fractionated craniospinal radiotherapy. AB - PURPOSE: Primary hypothyroidism is a common sequela of craniospinal radiotherapy in the treatment of childhood medulloblastoma. Due to the strong radiobiologic rationale, hyperfractionation can reduce the delayed effects of radiation injury. METHODS AND MATERIALS: The authors compared the incidence of thyroid dysfunction after conventionally fractionated radiotherapy (Group A, n = 20 patients) vs. hyperfractionated radiotherapy (Group B, n = 12 patients) in a group of pediatric patients with posterior fossa primitive neuroectodermal tumor (PNET). RESULTS: The mean age at the time of tumor diagnosis was 7.4 years in Group A and 8.4 years in Group B. Thyroid function was evaluated yearly, with ultrasonographic examination every 2 years. The patients were followed after diagnosis for a mean of 10.8 years for Group A and 6.0 years for Group B. Approximately 80% of the Group A (16/20) and 33.3% of the Group B (4/12) patients developed primary hypothyroidism within a similar period after irradiation (4.2 vs. 3.5 years, respectively). Analysis by cumulative incidence function demonstrated a significant difference in the risk of developing thyroid dysfunction between these two groups of patients (p < 0.05). Ultrasonography showed reduced thyroid volume in 7 Group A patients and structural changes in 21 patients (17 Group A, 4 Group B cases); a thyroid benign nodule was detected in 2 Group A patients. CONCLUSIONS: The current study findings suggest that the use of hyperfractionated craniospinal radiotherapy in the treatment of childhood medulloblastoma is associated with a lower risk of these patients' developing late thyroid dysfunction. PMID- 11483341 TI - Medication for hearing loss after fractionated stereotactic radiotherapy (SRT) for vestibular schwannoma. AB - PURPOSE: To investigate the effectiveness of corticosteroid treatments for patients showing decreases in hearing levels after stereotactic radiotherapy for vestibular schwannoma. METHODS AND MATERIALS: Twenty-one patients experienced a hearing loss in pure-tone average at greater than 20 dB or less than 10 dB within 1 year after irradiation administration of 44 Gy/22 fractions followed by a 4 Gy boost. Eight received oral prednisone at a daily dose of 30 mg, which was gradually decreased (medicated group), and 13 received none (nonmedicated group). The average observation period was 26.7 +/- 16.6 (range: 6--69) months. RESULTS: Hearing recovery was seen after initial onset of the hearing loss in all 8 patients in the medicated group and in 2 of 13 patients in the nonmedicated group (p = 0.001). The hearing recovery, that is, the change in pure-tone average (dB) at the last follow-up from the onset of hearing loss, was 9.8 +/- 6.9 dB (recovery) in the medicated group and -9.4 +/- 12.8 dB (further loss) in the nonmedicated group (p = 0.0013). The hearing recovery rate, normalizing to the degree of the hearing loss before medication, was also significantly higher in the medicated group than in the nonmedicated group (p = 0.0014). CONCLUSIONS: Corticosteroidal intake is suggested to be effective in improving hearing loss after stereotactic radiotherapy, at least in young patients having a useful pretreatment hearing level, if the treatment for hearing loss is administered immediately after the hearing loss is first detected. PMID- 11483342 TI - A phase I/II trial of three-dimensionally planned concurrent boost radiotherapy and protracted venous infusion of 5-FU chemotherapy for locally advanced rectal carcinoma. AB - BACKGROUND: Improving the response to preoperative therapy may increase the likelihood of successful resection of locally advanced rectal cancers. Historically, the pathologic complete response (pCR) rate has been < approximately 10% with preoperative radiation therapy alone and < approximately 20% with concurrent chemotherapy and radiation therapy. METHODS AND MATERIALS: Thirty-seven patients were enrolled on a prospective Phase I/II protocol conducted jointly at Washington University, St. Louis and the Catholic University of the Sacred Heart, Rome evaluating a three-dimensionally (3D) planned boost as part of the preoperative treatment of patients with unresectable or recurrent rectal cancer. Preoperative treatment consisted of 4500 cGy in 25 fractions over 5 weeks to the pelvis, with a 3D planned 90 cGy per fraction boost delivered once or twice a week concurrently (no time delay) with the pelvic radiation. Thus, on days when the boost was treated, the tumor received a dose of 270 cGy in one fraction while the remainder of the pelvis received 180 cGy. When indicated, nonaxial beams were used for the boost. The boost treatment was twice a week (total boost dose 900 cGy) if small bowel could be excluded from the boost volume, otherwise the boost was delivered once a week (total boost dose 450 cGy). Patients also received continuous infusion of 5-fluorouracil (1500 mg/m(2)-week) concurrently with the radiation as well as postoperative 5-FU/leucovorin. RESULTS: All 37 patients completed preoperative radiotherapy as planned within 32 -39 elapsed days. Twenty-seven underwent proctectomy; reasons for unresectability included persistent locally advanced disease (6 cases) and progressive distant metastatic disease with stable or smaller local disease (4 cases). Actuarial 3 year survival was 82% for the group as a whole. Among resected cases the 3-year local control and freedom from disease relapse were 86% and 69%, respectively.Twenty-four of the lesions (65%) achieved an objective clinical response by size criteria, including 9 (24%) with pCR at the primary site (documented T0 at surgery). The most important factor for pCR was tumor volume: small lesions with planning target volume (PTV) < 200 cc showed a 50% pCR rate (p = 0.02). There were no treatment associated fatalities. Nine of the 37 patients (24%) experienced Grade 3 or 4 toxicities (usually proctitis) during preoperative treatment. There were an additional 7 perioperative and 2 late toxicities. The most important factors for small bowel toxicity (acute or late) were small bowel volume (> or = 150 cc at doses exceeding 4000 cGy) and large tumor (PTV > or = 800 cc). For rectal toxicity the threshold is PTV > or = 500 cc. CONCLUSION: 3D planned boost therapy is feasible. In addition to permitting the use of nonaxial beams for improved dose distributions, 3D planning provides tumor and normal tissue dose-volume information that is important in interpreting outcome. Every effort should be made to limit the treated small bowel to less than 150 cc. Tumor size is the most important predictor of response, with small lesions of PTV < 200 cc most likely to develop complete responses. PMID- 11483343 TI - Local excision and postoperative radiotherapy for distal rectal cancer. AB - PURPOSE: To assess the outcome following local excision and postoperative radiotherapy (RT) for distal rectal carcinoma. MATERIALS AND METHODS: Seventy three patients received postoperative radiotherapy following local surgery for primary rectal carcinoma at Princess Margaret Hospital from 1983 to 1998. Selection factors for postoperative RT were patient preference, poor operative risks, and "elective" where conservative therapy was regarded as optimal therapy. Median distance of the primary lesion from the anal verge was 4 cm (range, 1--8 cm). There were 24 T1, 36 T2, and 8 T3 lesions. The T category could not be determined in 5. Of 55 tumor specimens in which margins could be adequately assessed, they were positive in 18. RT was delivered using multiple fields by 6- to 25-MV photons. Median tumor dose was 50 Gy (range, 38--60 Gy), and 62 patients received 50 Gy in 2.5-Gy daily fractions. The tumor volume included the primary with 3--5 cm margins. No patients received adjuvant chemotherapy. Median follow up was 48 months (range, 10--165 months). RESULTS: Overall 5-year survival and disease-free survival were 67% and 55%, respectively. Tumor recurrence was observed in 23 patients. There were 14 isolated local relapses; 6 patients developed local and distant disease; and 3 relapsed distantly only. For patients with T1, T2, and T3 lesions, 5-year local relapse-free rates were 61%, 75%, and 78%, respectively, and 5-year survival rates were 76%, 58%, and 33%, respectively. The 5-year local relapse-free rate was lower in the presence of lymphovascular invasion (LVI) compared to no LVI, 52% vs. 89%, p = 0.03, or where tumor fragmentation occurred during local excision compared to no fragmentation, 51% vs. 76%, p = 0.02. Eleven of 14 patients with local relapse only underwent abdominoperineal resection, 8 achieved local control, and 4 remained cancer free. The ultimate local control, including salvage surgery, was 86% at 5 and 10 years. The 5-year colostomy-free rate was 82%. There were 2 patients who experienced RTOG Grade 3 late complications, and 1 with Grade 4 complication (bowel obstruction requiring surgery). CONCLUSION: The local relapse rate for patients with T1 disease was high compared to other series of local excision and postoperative RT. Patients with LVI or tumor fragmentation during excision have high local relapse rates and may not be good candidates for conservative surgery and postoperative RT. PMID- 11483344 TI - Phase I study of twice-weekly gemcitabine and concomitant external-beam radiotherapy in patients with adenocarcinoma of the pancreas. AB - PURPOSE: To determine the maximum tolerated dose and dose-limiting toxicity associated with twice-weekly gemcitabine and concomitant external-beam radiotherapy in patients with adenocarcinoma of the pancreas. METHODS AND MATERIALS: Twenty-one patients with biopsy-proven adenocarcinoma of the pancreas were treated with external-beam radiotherapy to a dose of 50.4 Gy in 28 fractions, concurrent with gemcitabine, infused over 30 min before irradiation on a Monday and Thursday schedule. The dose of gemcitabine was escalated in 5 cohorts of 3--6 patients each. Initial gemcitabine dose was 10 mg/m(2), with dose escalation until dose-limiting toxicity was observed. RESULTS: The maximum tolerated dose of gemcitabine was 50 mg/m(2), when given in a twice-weekly schedule with radiation. Dose-limiting toxicity was seen in 2 patients at 60 mg/m(2), and consisted of severe upper gastrointestinal bleeding approximately 1 month after completion of treatment. Six patients had radiographic evidence of response to treatment, and 5 of these underwent complete surgical resection. Three patients who underwent complete resection had been deemed to have unresectable tumors before enrollment on trial. Four patients are alive, including 2 without evidence of disease more than 1 year after resection. CONCLUSION: The combination of external-beam radiation and twice-weekly gemcitabine at a dose of 50 mg/m(2) is well tolerated and shows promising activity for the treatment of pancreatic cancer. Our data suggest a higher maximum tolerated dose and different dose-limiting toxicity than previously reported. Further investigation of this regimen is warranted. PMID- 11483345 TI - Novel DNA sequence variants in the hHR21 DNA repair gene in radiosensitive cancer patients. AB - PURPOSE: Radiation therapy is an important treatment modality for oncology patients. DNA sequence variants have so far been identified in only a few genes in radiosensitive cancer patients. Patients known to be clinically radiosensitive were tested for mutation of a gene involved in DNA double-strand break repair and sister chromatid cohesion--hHR21. METHODS AND MATERIALS: Clinically radiation sensitive patients were accrued to the study after giving informed consent. Blood samples were obtained and lymphoblastoid cell lines established. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to amplify the hHR21 gene, and the DNA product was sequenced to identify any genetic abnormalities. Northern blot analysis, cell survival, and growth assays were performed on control cells and cells with hHR21 variants, and a restriction digest assay was developed to screen for carriers of a detected gene variant. RESULTS: The DNA sequence of the hHR21 gene was determined in 19 radiation sensitive cancer patients. In 6 of the 19 patients, a thymidine (T) to cytosine (C) transition was detected at position 1440 of the hHR21 open reading frame (T1440C). This variant did not alter the amino acid sequence and was likely to be a polymorphism. One patient with a particularly severe radiation reaction had a second sequence variant immediately adjacent to the first. This was a guanine (G) to adenine (A) transition (G1441A), resulting in a change of the amino acid sequence (glycine --> arginine) in a portion of the protein conserved in evolution. This suggests that this DNA alteration may be biologically significant. Restriction digest with the HpaII enzyme confirmed the presence of both sequence variants on the same allele. CONCLUSIONS: We describe the first two DNA sequence variants ever found in the hHR21 gene, in patients with clinical radiation hypersensitivity. Although no direct evidence for the involvement of hHR21 alterations in the radiosensitivity of the cancer patients examined has been demonstrated, the possibility exists that homozygous mutations or other mutations of this gene could contribute to radiosensitivity. A simple test is described that could be applied to screening for these variants in relevant populations. PMID- 11483346 TI - Radiation-induced glomerular thrombus formation and nephropathy are not prevented by the ADP receptor antagonist clopidogrel. AB - PURPOSE: To assess the effects of kidney irradiation on glomerular adenosine diphosphatase (ADPase) activity and intraglomerular microthrombus formation, and their correlation to the development of renal functional impairment. METHODS AND MATERIALS: C3H/HenAf-nu(+) mice were given single-dose or fractionated kidney irradiations. Glomerular ADPase activity was measured using a cerium-based histochemical method. Microthrombus formation within the glomeruli was assessed by a semiquantitative immunohistochemical analysis of fibrinogen/fibrin deposits. Renal function was assessed by the [(51)Cr]EDTA retention assay. RESULTS: The ADPase activity was significantly reduced, to approximately 50% of pretreatment value, 4--40 weeks after 10--16 Gy single-dose irradiation and at 44 weeks after 20 x 2 Gy. No dose--effect relationship was found. An approximately fourfold increase in glomerular fibrinogen/fibrin staining was observed at 1 year after irradiation. This increase was not influenced by treating the mice with daily, oral clopidogrel, a platelet ADP receptor antagonist, which reduced platelet aggregation by more than 75%. Radiation-induced impairment of glomerular filtration was also not affected by the clopidogrel treatment. CONCLUSION: These data indicate that irradiation significantly reduced glomerular ADPase activity, which correlated with an increased glomerular fibrinogen/fibrin deposition. We were not able to reduce these prothrombotic changes, nor to protect against radiation nephropathy, by pharmacological intervention with an ADP-receptor antagonist. PMID- 11483347 TI - Monte Carlo evaluation of tissue inhomogeneity effects in the treatment of the head and neck. AB - PURPOSE: To use Monte Carlo dose calculation to assess the degree to which tissue inhomogeneities in the head and neck affect static field conformal, computed tomography (CT)-based 6-MV photon treatment plans. METHODS AND MATERIALS: We retrospectively studied the three-dimensional treatment plans that had been used for the treatment of 5 patients with tumors in the nasopharyngeal or paranasal sinus regions. Two patients had large surgical cavities. The plans were designed with a clinical treatment planning system that uses a measurement-based pencil beam dose-calculation algorithm with an equivalent path-length inhomogeneity correction. Each plan employs conformally-shaped 6-MV photon beams. Patient anatomy and electron densities were obtained from the treatment planning CT images. For each plan, the dose distribution was recalculated with the Monte Carlo method, utilizing the same beam geometry and CT images. The Monte Carlo method accurately accounts for the perturbation effects of local tissue heterogeneities. The Monte Carlo calculated dose distributions were compared with those from the clinical treatment planning system. RESULTS: The degree to which tissue inhomogeneity affects the dose distributions of individual fields varies with the specific anatomic geometry, especially the size and location of air cavities in relation to the beam orientation and field size. Most of the beam apertures completely enclose the air cavities within or adjacent to the gross tumor volume (GTV). Equivalent squares (including blocking) ranged from approximately 5 to 9.5 cm. A common feature observed for individual fields is that the Monte Carlo calculated doses to tissue directly behind and within an air cavity are lower. However, after combining the fields employed in each treatment plan, the overall dose distribution shows only small differences between the two methods. For all 5 patients, the Monte Carlo calculated treatment plans showed a slightly lower dose received by the 95% of target volume (D(95)) than the plans calculated with the pencil-beam algorithm. The average difference in the target volume encompassed by the prescription isodose line was less than 2.2%. The difference between the dose-volume histograms (DVHs) of the GTV was generally small. For the brainstem and chiasm, the DVHs of the two plans were similar. For the spinal cord, differences in the details of the DHV and the dose to 1 cc (D(1cc)) of the structure were observed, with Monte Carlo calculation generally predicting increased dose indices to the spinal cord. However, these changes are not expected to be clinically significant. CONCLUSION: For 6-MV photons, the effects of both normal tissue inhomogeneities and surgical air cavities on the target coverage were adequately accounted for by conventional pencil beam methods for all of the cases studied. Although differences in details of the DVHs of the normal structures were observed, depending on whether Monte Carlo or pencil-beam algorithm was used for calculation, these differences are not expected to be clinically significant. In general, the pencil-beam calculation corrected for primary attenuation by the equivalent pathlength is a sufficiently accurate method for head-and-neck treatment planning using 6-MV photons. PMID- 11483348 TI - A protocol for the reduction of systematic patient setup errors with minimal portal imaging workload. AB - PURPOSE: To evaluate a new off-line patient setup correction protocol that minimizes the required number of portal images and perform a comparison with currently applied protocols. METHODS AND MATERIALS: We compared two types of off line protocols: (a) the widely applied shrinking action level (SAL) protocol, in which the setup error, averaged over the measured treatment fractions, is compared with a threshold that decreases with the number of measurements, to decide if a correction is necessary; and (b) a new "no-action-level" (NAL) protocol, which simply calculates the mean setup error over a fixed number of fractions, and always corrects for it. The performance of the protocols was evaluated by applying them to (a) a database of measured setup errors from 600 prostate patients (with, on average, 10 imaged fractions/patient) and (b) Monte Carlo-generated setup error distributions for various values of the population systematic and random errors. RESULTS: The NAL protocol achieved a significantly higher accuracy than the SAL protocol for a similar workload in terms of image acquisition and analysis, as well as in setup corrections. The SAL protocol required approximately three times more images than the NAL protocol to obtain the same reduction of systematic errors. Application of the NAL protocol to measured setup errors confirmed its efficacy in systematic error reduction in a real patient population. CONCLUSION: The NAL protocol performed much more efficiently than the SAL protocol for both actually measured and simulated setup data. The resulting decrease in required portal images not only reduces workload, but also dose to healthy tissue, if dedicated large fields are required for portal imaging (double exposure). PMID- 11483349 TI - Variability in target volume delineation on CT scans of the breast. AB - PURPOSE: To determine the intra- and interobserver variation in delineation of the target volume of breast tumors on computed tomography (CT) scans in order to perform conformal radiotherapy. MATERIALS AND METHODS: The clinical target volume (CTV) of the breast was delineated in CT slices by four radiation oncologists on our clinically used delineation system. The palpable glandular breast tissue was marked with a lead wire on 6 patients before CT scanning, whereas 4 patients were scanned without a lead wire. The CTV was drawn by each observer on three separate occasions. Planning target volumes (PTVs) were constructed by expanding the CTV by 7 mm in each direction, except toward the skin. The deviation in the PTV extent from the average extent was quantified in each orthogonal direction for each patient to find a possible directional dependence in the observer variations. In addition, the standard deviation of the intra- and interobserver variation in the PTV volume was quantified. For each patient, the common volumes delineated by all observers and the smallest volume encompassing all PTVs were also calculated. RESULTS: The patient-averaged deviations in PTV extent were larger in the posterior (42 mm), cranial (28 mm), and medial (24 mm) directions than in the anterior (6 mm), caudal (15 mm), and lateral (8 mm) directions. The mean intraobserver variation in volume percentage (5.5%, 1 SD) was much smaller than the interobserver variation (17.5%, 1 SD). The average ratio between the common and encompassing volume for the four observers separately was 0.82, 0.74, 0.82, and 0.80. A much lower combined average ratio of 0.43 was found because of the large interobserver variations. For the observer who placed the lead wire, the intraobserver variation in volume was decreased by a factor of 4 on scans made with a lead wire in comparison to scans made without a lead wire. For the other observers, no improvement was seen. Based on these results, an improved delineation protocol was designed. CONCLUSIONS: Intra- and especially interobserver variation in the delineation of breast target volume on CT scans can be rather large. A detailed delineation protocol making use of CT scans with lead wires placed on the skin around the palpable breast by the delineating observer reduces the intraobserver variation. To reduce the interobserver variation, better imaging techniques and pathology studies relating glandular breast tissue to imaging may be needed to provide more information on the extent of the clinical target volume. PMID- 11483351 TI - Effects of litter size on maternal care, body weight and infant development in golden hamsters (Mesocricetus auratus). AB - The effects of litter size on maternal care, body weight and infant development of golden hamsters were investigated from a longitudinal perspective. Litters were culled to 1,3,6 and 9 pups, and the behavior and body weight of mothers and pups were recorded from the 5th to the 25th postpartum day. We noted that the time spent by mothers in bodily interactions with pups decreased as a function of litter size; maternal pup retrievals reached their maximum around the 13-15th day, which coincided with the increased locomotor activity of pups at this time; the total number of pup retrievals by the mother increased as a function of the litter size, but mothers of larger litters were more 'efficient' (i.e. they failed less frequently in exhibiting a full sequence of retrievals) and exhibited a low litter-size proportional mean number of retrievals. All mothers gradually lost body mass throughout lactation, and decrease in body weight was significantly related to litter size. The mean body weight gain (%) by pups decreased as a function of litter size, but we also noted that single and larger litter pups exhibited a decreased body mass (grams) by the 15th day, suggesting that infant development may be impaired at both extremes of experimental conditions. We concluded that the behavior of mothers and pups was affected by the litter size, and it appeared that the litter had an optimal size-not so large as to overlap the mother's physical capacity, and not so small as to fail to compensate for the parental investment. PMID- 11483352 TI - Responsiveness to brightness change in hooded rats: effects of sex and procedure. AB - In an investigation of recognition memory involving a preference test, hooded rats of both sexes were individually confined to the stem and choice area of a T- or Y-maze by means of clear Perspex barriers across each arm entrance that enabled the subjects to see into but not enter the arms. Following removal of the barriers and changing of one arm to opposite brightness, the first arm entered and the number of entries of and time spent in each arm were recorded. In the first experiment, rats entered first the arm that had been changed. During the first minute of observation, they also entered this novel arm more often and spent more time in it than the unchanged arm, irrespective of the type of change. In a second experiment, when the change was from one arm black and the other white to two black arms, more responsiveness occurred after 6-min prior exposure (without access) than after 3 min. In both experiments, the nature of the apparatus (T- or Y-maze) affected several outcomes, but the most significant influence was of the sex of the subjects. Females appeared less responsive to change than males as determined by entries of and time spent in the changed arm. Rather than inferiority of females in recognition or spatial memory, the sex effects were most likely due to their more rapid habituation to novelty possibly assisted by superior visual exploration capacities. PMID- 11483353 TI - Adult female prairie voles and meadow voles do not suppress reproduction in their daughters. AB - Reproductive suppression of young females by conspecific females has been reported from laboratory studies on several species of rodents, including the prairie vole, Microtus ochrogaster, but not meadow voles, M. pennsylvanicus. We exposed female prairie voles and meadow voles to two treatments: a mother and one 23-26-day-old daughter paired with a strange male and a 23-26-day-old daughter paired with a different strange male. We found no differences in the proportion of daughters breeding or the time to sexual maturation for daughters raised in the two treatments for either species. Thus, we have no indication that mothers had any adverse effect on reproductive efforts of their daughters. These results differ from previous studies that concluded young female prairie voles were reproductively suppressed by female relatives. The difference between our and previous studies on reproductive suppression is that we examined breeding in young females rather than proximate measures of growth and reproductive development. We question the evolutionary significance of reproductive suppression among related female microtine rodents, especially in that it has not been documented from field populations. PMID- 11483354 TI - Within-session changes in responding during concurrent variable interval variable ratio schedules. AB - Rats (Experiment 1) and pigeons (Experiment 2) responded on several concurrent variable interval (VI) variable ratio (VR) schedules. The rate of, but not the time spent, responding in each component usually changed within-sessions. The bias and sensitivity parameters of the generalized matching law (GML) did not change systematically within-sessions. The fit of the GML to the data did not change within-sessions for pigeons, but it was better in the middle than at the beginning or end of the session for some for rats. Both over- and under-matching occurred. These results imply that within-session changes in responding do not usually cause problems for assessing the validity of the GML when subjects respond on concurrent VI VR schedules. The results also suggest that under- and over-matching are not produced by different factors, but rather lie on a continuum. PMID- 11483355 TI - The molecular biology and nomenclature of the activating transcription factor/cAMP responsive element binding family of transcription factors: activating transcription factor proteins and homeostasis. AB - The mammalian ATF/CREB family of transcription factors represents a large group of basic region-leucine zipper (bZip) proteins which was originally defined in the late 1980s by their ability to bind to the consensus ATF/CRE site 'TGACGTCA'. Over the past decade, cDNA clones encoding identical or homologous proteins have been isolated by different laboratories and given different names. These proteins can be grouped into subgroups according to their amino acid similarity. In this review, we will briefly describe the classification of these proteins with a historical perspective of their nomenclature. We will then review three members of the ATF/CREB family of proteins: ATF3, ATF4 and ATF6. We will address four issues for each protein: (a) homologous proteins and alternative names, (b) dimer formation with other bZip proteins, (c) transcriptional activity, and (d) potential physiological functions. Although the name Activating Transcription Factor (ATF) implies that they are transcriptional activators, some of these proteins are transcriptional repressors. ATF3 homodimer is a transcriptional repressor and ATF4 has been reported to be either an activator or a repressor. We will review the reports on the transcriptional activities of ATF4, and propose potential explanations for the discrepancy. Although the physiological functions of these proteins are not well understood, some clues can be gained from studies with different approaches. When the data are available, we will address the following questions. (a) How is the expression (at the mRNA level or protein level) regulated? (b) How are the transcriptional activities regulated? (c) What are the interacting proteins (other than bZip partners)? (d) What are the consequences of ectopically expressing the gene (gain-of-function) or deleting the gene (loss-of-function)? Although answers to these questions are far from being complete, together they provide clues to the functions of these ATF proteins. Despite the diversity in the potential functions of these proteins, one common theme is their involvement in cellular responses to extracellular signals, indicating a role for these ATF proteins in homeostasis. PMID- 11483356 TI - Identification and comparative analysis of a second runx3 promoter. AB - The runx gene family encodes transcriptional regulators that are essential in several processes. Alternative transcription start sites (3' or P1 and 5' or P2) have been described both at runx1 and runx2. Functional domains have been mapped to P2 N-termini. Here, starting from an analysis of cDNA clones, we identify a P2 promoter at mouse runx3. In functional assays in COS-7 cells, this promoter is as active as runx1 P2. By nuclease protection, primer extension and deletion analysis we map a major start site and define a minimal promoter element. The activity of both promoters at all three mouse loci was systematically compared in a comprehensive panel of fresh tissues. The conservation of a two-promoter structure and their distinctive pattern of activity support the hypothesis that alternative 5' isoforms subserve unique functions. PMID- 11483357 TI - Cloning of peroxisomal ascorbate peroxidase gene from barley and enhanced thermotolerance by overexpressing in Arabidopsis thaliana. AB - A full-length cDNA clone (HvAPX1) encoding a peroxisomal type ascorbate peroxidase was isolated from barley (Hordeum vulgare cv. Haruna-nijyo) leaves by differential display. The deduced amino acid sequence of the HvAPX1 gene had 75.3% homology to that from the Gossypium hirsutum glyoxysomal APX gene and 72.1% homology to that from the Arabidopsis thaliana peroxisomal APX gene, APX3. Southern blot analysis indicated that a single-copy gene in the barley genome encoded HvAPX1. Northern blot analysis showed that the HvAPX1 transcript increased remarkably in response to heat, salt and abscisic acid treatment. Induction was not caused by treatment with hydrogen peroxide. The HvAPX1 gene was introduced into A. thaliana under control of the 35S RNA promoter of the cauliflower mosaic virus. The transgenic plants were significantly more tolerant to heat stress as compared with the wild-type. PMID- 11483358 TI - Differential expression and subcellular distribution of the mouse metastasis associated proteins Mta1 and Mta3. AB - The human metastasis-associated gene (MTA1) is overexpressed in cell lines and tissues representing metastatic tumors. Here we report cloning of the mouse Mta1 as well as a novel structurally related mouse gene, Mta3. The mouse Mta1 protein shares 94 and 59% homology to the human MTA1 and mouse Mta3 proteins, respectively. Northern blotting analysis using an Mta1 cDNA probe revealed a prevalent 3 kb hybridization signal in all mouse tissues except the skeletal muscle while a smaller approximately 1.0 kb mRNA product was also detected in the heart. Mta3 transcripts (approximately 2 kb) were detected in most tissues with an additional approximately 6.2 kb signal detected in the brain. In vitro transcription/translation of the full-length Mta1 and Mta3 cDNAs generated products of the expected molecular masses, i.e. 80 and 60 kDa, respectively. To assess subcellular localization, green fluorescence protein (GFP)-tagged expression constructs of Mta1 and Mta3 and various deletion constructs of GFP Mta1 were transiently expressed in Balb/MK keratinocytes. GFP-Mta1 was found exclusively in the nucleus while GFP-Mta3 was present in both the nucleus and cytoplasm. Compared to Mta3, the carboxy terminal end of Mta1 contains an additional nuclear localization signal (NLS) and a proline-rich Src homology 3 (SH3) ligand. The results of transient expression experiments of various Mta1 fragments containing these domains in different combinations indicated that nuclear localization of Mta1 depended on the presence of at least one NLS and one SH3 binding site. These SH3 ligands appeared to be functional as they facilitated interaction with the adaptor protein, Grb2, and the Src-family tyrosine kinase, Fyn. PMID- 11483359 TI - Two-domain hemoglobin gene of the water flea Moina macrocopa: duplication in the ancestral Cladocera, diversification, and loss of a bridge intron. AB - Two cDNAs encoding the two-domain hemoglobin (Hb) chains of a crustacean Cladocera, Moina macrocopa, were cloned and their nucleotide (nt) sequences were determined. The amino acid (aa) sequences of both the gene products deduced from the nt sequences consisted of 348 residues and showed 98% identity with each other. These sequences together with the NH(2)-terminal aa sequences of the Hb chains determined after separation by two-dimensional gel electrophoresis showed that the Hb chains are synthesized as a secretory precursor with a signal peptide of 17 aa residues. The aa sequences of M. macrocopa Hb chains shared the following features with those of Daphnia Hb chains. Firstly, the signal peptide is followed by an NH(2)-terminal extension containing a threonine-rich sequence that might play a role in the multimerization of subunit chains. Secondly, the identity between the aa sequences of the first and second domains is exceptionally low. These facts suggest that duplication of the cladoceran Hb gene occurred before the divergence of families Moinidae and Daphniidae. Analysis of genomic DNA showed that the M. macrocopa Hb genes consist of two large repeated regions, encoding the first and second domains of Hb chains, respectively. The intron-exon organization of the first region of the M. macrocopa Hb genes was similar to that found in the Daphnia Hb genes, having the three-exon, two-intron structure characteristic of animal Hb genes. However, the intron bridging the two regions and the most downstream intron in the second region were missing in the Moina genes, providing a new example of intron loss. The following elements in the 5' flanking region were conserved in the Moina and Daphnia genes: (1) TATAAA, a typical TATA box sequence accompanied by a downstream sequence, GAAXAGCATCAGTT (the fourth residue X was G or A in Daphnia and absent in Moina); (2) CCAAT boxes, located upstream of the TATA box; (3) the binding sites for HIF-1 and GATA 1, also located upstream of the TATA box, that may be responsible for up regulation of the cladoceran Hb genes under hypoxia. PMID- 11483360 TI - Full-sized HERV-K (HML-2) human endogenous retroviral LTR sequences on human chromosome 21: map locations and evolutionary history. AB - One of the evolutionary mechanisms for acquisition of novel functional sequences can be domestication of exogenous retroviruses that have been integrated into the germ line. The whole genome mapping of such elements in various species could reveal differences in positions of the retroviral integration and suggest possible roles of these differences in speciation. Here, we describe the number, locations and sequence features of the human endogenous retrovirus HERV-K (HML-2) long terminal repeat (LTR) sequences on human chromosome 21. We show that their distribution along the chromosome is not only non-random but also roughly correlated with the gene density. Amplification of orthologous LTR sites from a number of primate genomes produced patterns of presence and absence for each LTR sequence and allowed determination of the phylogenetic ages and evolutionary order of appearance of individual LTRs. The identity level and phylogenetic age of the LTRs did not correlate with their map locations. Thus, despite the non random distribution of LTRs, they have apparently been inserted randomly into the chromosome relative to each other. As evidenced in previous studies of chromosomes 19 and 22, this is a characteristic of HERV-K integration. PMID- 11483361 TI - Gene expressivity is the main factor in dictating the codon usage variation among the genes in Pseudomonas aeruginosa. AB - Codon usage biases of all DNA sequences (length greater than or equal to 300 bp) from the complete genome of Pseudomonas aeruginosa have been analyzed. As P. aeruginosa is a GC-rich organism, G and/or C are expected to predominate in their codons. Overall codon usage data analysis indicates that indeed codons ending in G and/or C are predominant in this organism. But multivariate statistical analysis indicates that there is a single major trend in the codon usage variation among the genes in this organism, which has a strong negative correlation with the expressivities of the genes. The majority of the lowly expressed genes are scattered towards the positive end of the major axis whereas the highly expressed genes are clustered towards the negative end. This is the first report where the prokaryotic organism having highly skewed base composition is dictated mainly by translational selection, though some other factors such as the lengths of the genes as well as the hydrophobicity of genes also influence the codon usage variation among the genes in this organism in a minor way. PMID- 11483362 TI - Structural organization of the rat mitogen-activated protein kinase phosphatase 2 gene. AB - Mitogen-activated protein kinase (MAPK) phosphatases (MKPs) are dual specificity protein phosphatases that specifically inactivate MAPKs. Regulated expression of MKPs plays a key role in determining their physiological function. However, little is known about the molecular mechanism of the activation of MKP genes. In this study, we cloned the rat MKP-2 gene and characterized its structure. The MKP 2 gene has four exons and three introns. The organization of exons of the MKP-2 gene is very similar to that of the MKP-1 gene, suggesting that MKP-1 and MKP-2 are derived from the same ancestral gene. We identified multiple transcription start sites (TSSs) for the MKP-2 gene. There is no functional TATA motif in the 5' proximal region of the TSSs. Instead, this region is highly GC-rich and has two putative Sp1 sites. A 1.8 kb 5' flanking region of the MKP-2 gene is sufficient to mediate transcriptional activation of the luciferase reporter gene by phorbol ester in GH3 cells. These results provide essential information about structural organization and regulatory sequences of the MKP-2 gene for further investigation of the molecular mechanisms of MKP-2 induction by extracellular stimuli. PMID- 11483363 TI - Molecular cloning and characterization of canine metalloproteinase-9 gene promoter. AB - This paper describes the cloning and characterization of the canine matrix metalloproteinase-9 (MMP-9) gene promoter. The 5' untranslated region was obtained by genome walking upstream of the canine MMP-9 translation start site using canine genomic DNA as template. A DNA fragment of 1894 bp was isolated and on analysis demonstrated regions of sequence homology with the MMP-9 promoter sequences already determined for other species. In general, conserved regions correlated with DNA binding motifs such as a TATA-like box, AP-1 sites, GC boxes and a nuclear factor-kappaB binding domain. The DNA promoter fragment was sufficient to drive basal expression of a luciferase reporter gene in Madin Darby canine kidney (MDCK) cells and to a lesser extent in feline embryonic fibroblast (FEA) cells. Activity of the promoter was enhanced by the treatment of transfected MDCK cells with phorbol 12-myristate 13-acetate but no effect was observed in the FEA cells. Promoter deletion studies revealed that regions of promoter were necessary for induction of reporter gene expression. PMID- 11483364 TI - Two new genes from the human ATP-binding cassette transporter superfamily, ABCC11 and ABCC12, tandemly duplicated on chromosome 16q12. AB - Several years ago, we initiated a long-term project of cloning new human ATP binding cassette (ABC) transporters and linking them to various disease phenotypes. As one of the results of this project, we present two new members of the human ABCC subfamily, ABCC11 and ABCC12. These two new human ABC transporters were fully characterized and mapped to the human chromosome 16q12. With the addition of these two genes, the complete human ABCC subfamily has 12 identified members (ABCC1-12), nine from the multidrug resistance-like subgroup, two from the sulfonylurea receptor subgroup, and the CFTR gene. Phylogenetic analysis determined that ABCC11 and ABCC12 are derived by duplication, and are most closely related to the ABCC5 gene. Genetic variation in some ABCC subfamily members is associated with human inherited diseases, including cystic fibrosis (CFTR/ABCC7), Dubin-Johnson syndrome (ABCC2), pseudoxanthoma elasticum (ABCC6) and familial persistent hyperinsulinemic hypoglycemia of infancy (ABCC8). Since ABCC11 and ABCC12 were mapped to a region harboring gene(s) for paroxysmal kinesigenic choreoathetosis, the two genes represent positional candidates for this disorder. PMID- 11483365 TI - The TGV transgenic vectors for single-copy gene expression from the Escherichia coli chromosome. AB - Plasmid-based cloning and expression of genes in Escherichia coli can have several problems: plasmid destabilization; toxicity of gene products; inability to achieve complete repression of gene expression; non-physiological overexpression of the cloned gene; titration of regulatory proteins; and the requirement for antibiotic selection. We describe a simple system for cloning and expression of genes in single copy in the E. coli chromosome, using a non antibiotic selection for transgene insertion. The transgene is inserted into a vector containing homology to the chromosomal region flanking the attachment site for phage lambda. This vector is then linearized and introduced into a recombination-proficient E. coli strain carrying a temperature-sensitive lambda prophage. Selection for replacement of the prophage with the transgene is performed at high temperature. Once in the chromosome, transgenes can be moved into other lysogenic E. coli strains using standard phage-mediated transduction techniques, selecting against a resident prophage. Additional vector constructs provide an arabinose-inducible promoter (P(BAD)), P(BAD) plus a translation initiation sequence, and optional chloramphenicol-, tetracycline-, or kanamycin resistance cassettes. These Transgenic E. coli Vectors (TGV) allow drug-free, single-copy expression of genes from the E. coli chromosome, and are useful for genetic studies of gene function. PMID- 11483366 TI - Tissue distribution of porcine peroxisome proliferator-activated receptor alpha: detection of an alternatively spliced mRNA. AB - Peroxisome proliferator-activated receptor alpha (PPAR alpha) plays a key role in regulating the catabolic pathway of lipids in response to a variety of compounds named peroxisome proliferators (PPs). The cellular responses to PPs differ among mouse/rat and other species and actualize the study in swine, which show close resemblance to human lipid physiology and metabolism. We have isolated the cDNA containing the open reading frame of porcine PPAR alpha whose deduced amino acid sequence revealed an evolutionary distance to mouse/rat that could be implicated in causing the species-dependent response to PPs. Interestingly, an alternatively spliced PPAR alpha mRNA, lacking exon 5, was detected by reverse transcriptase polymerase chain reaction in several porcine tissues. This deletion alters the reading frame and introduces a premature stop codon of PPAR alpha, presumably giving rise to a C-terminal truncated protein. We have also examined PPAR alpha expression by Northern blot analysis in tissues taken from pigs at three different stages of maturation, including two breeds that differ considerably in body composition and fat deposition. Porcine PPAR alpha was predominantly expressed in kidney and liver in mature individuals. When comparing piglets of a young age, a breed-specific tissue distribution of PPAR alpha mRNA was observed, particularly in liver and heart. PMID- 11483367 TI - The complete sequence of the human locus for NgCAM-related cell adhesion molecule reveals a novel alternative exon in chick and man and conserved genomic organization for the L1 subfamily. AB - NrCAM is one member of the L1 subfamily of cell surface recognition molecules implicated in nervous system development and function. Here we report the complete sequence of the human NRCAM locus. The gene comprises 34 exons and shows extensive conservation of exon/intron structure compared to L1, suggesting a common evolutionary ancestor. By human-chick sequence comparison we identified exons not previously found in mammalian NRCAM mRNAs. One of these encodes a premature stop codon that would give rise to an isoform of NRCAM lacking ankyrin binding capacity. The availability of the complete sequence will allow an investigation of the potential role of these splice variants, and examination of the regulatory elements controlling NRCAM expression as well as the relationship of NRCAM to disorders involving 7q. PMID- 11483368 TI - Cloning and characterization of a gene coding for a novel metallothionein in the Pacific oyster Crassostrea gigas (CgMT2): a case of adaptive response to metal induced stress? AB - Cases of heavy metal resistance acquisition have already been demonstrated in eukaryotes, which involve metallothionein (MT) gene duplication or amplification mechanisms. We characterized in a marine bivalve, Crassostrea gigas, a gene coding for an unusual MT, which has never been described in other species. Our results illustrate a unique case of exon duplication and rearrangement in the MT gene family. The particular organization of the third exon of this gene allows the synthesis of a MT that presents a higher metal ion binding capacity compared to previously described MTs. The formation of a supplementary third structural beta-domain is proposed to explain results obtained in in vitro experiments. Differences in the metal responsive element (MRE) copy number and MRE core sequence observed in the promoter of CgMT2 also suggest differential regulation of CgMT2 transcription and possible implication in the detoxification processes. PMID- 11483369 TI - Molecular cloning, expression and regulation of the avian tubby-like protein 1 (tulp1) gene. AB - The tubby-like protein 1 (tulp1) gene is a member of the tubby multigene family which includes tub, tulp1, tulp2 and tulp3. Human and mouse tulp1 genes were cloned and mutations in tulp1 have been implicated in retinitis pigmentosa in man. Here we report on the cDNA cloning of the chicken tulp1 homologue and its protein product deduced from the nucleotide sequence. The chicken Tulp1 protein comprises 358 amino acids with a calculated molecular mass of 40 kDa. The overall structure of Tub and Tulp proteins, exemplified by the highly conserved C terminal domain of 255 amino acids and the signature motif KLACE, is also preserved in chicken Tulp1. Phylogenetic analysis demonstrates that chicken tulp1 cDNA and protein are closely related to human and mouse tulp1. In addition, chicken tulp1 mRNA is abundantly expressed in retina similar to tulp1 expression in human and mouse. Two tulp1-specific transcripts of 2 and 3 kb in size were identified that showed differential regulation during embryonic and postnatal development. Finally, tulp1 mRNA was found to be expressed in chicken erythroid cells and upregulated by ligand-activated thyroid hormone receptor (TR alpha/c erbA). PMID- 11483370 TI - Gastric antiulcer effect of Rhizophora mangle L. AB - The effects of the freeze-dried aqueous extract from red mangrove bark on gastric ulceration induced by ethanol-hydrochloric acid were studied in rats. Mucosal damage was compared with that given with cimetidine. The effects of these agents on the quantity and quality of the gastric mucus were also determined. Oral treatment with red mangrove extract at 500 mg/kg body weight (b.w.) gave the highest level of gastric protection. Mucus content was increased and it was accompanied by a proportional increase in proteins. The group, which received cimetidine, showed no effect on the mucus secretion induced in this experimental model. PMID- 11483371 TI - An ethnobotanical study of plants used for the treatment of sexually transmitted diseases (njovhera) in Guruve District, Zimbabwe. AB - The use of medicinal plants in Guruve District, Zimbabwe, contributes significantly to primary health care of the people of the area. This paper presents findings of a survey of plants used for the treatment of sexually transmitted diseases (STDs) in the district. Ethnobotanical information obtained from traditional herbalists and other knowledgeable rural dwellers, has revealed 15 plant species belonging to 10 families as medicinal plants used for the treatment of these infections in the area. Six of these are the commonest and most frequently prescribed by the healers. Roots are the most frequently used parts of the plants constituting 53% of preparations while oral administration of extracts is the main method of prescription. Based on the information gathered from the traditional healers, Acacia nilotica (L.) Willd. ex Delile, Cassia abbreviata Oliv. Dichrostachys cinerea Wight and Arn, Solanum incanum L., Vernonia amygdalina Del. and Zanha africana (Radlk) Excell. are the most frequently used plants for the treatment of STDs. The methanol extracts of Cassia abbreviata, Zanha africana and Acacia nilotica showed significant inhibition against Gram-positive and Gram-negative bacteria, while acetone extracts of these plants inhibited most of the species. Generally the water extracts show less activity than acetone and methanol extracts. PMID- 11483372 TI - Effect of a herbal protein CI-1, purified from Cajanus indicus on the ultrastructural study of hepatocytes, in models of liver failure in mice. AB - Ultrastructural changes in acute liver damage models in swiss albino mice (male, 30 g +/-2) induced by CCl(4) (0.1 ml/100 g); beta-galactosamine (500 mg/kg); paracetamol (300-500 mg/kg) and 40% ethanol (2 ml/100 g) were studied. Electron microscopical studies of hepatocytes of treated (hepatotoxins) mice showed dilation of ER of both rough and smooth type with swollen mitochondria. Ethanol treated mouse hepatocytes showed giant mitochondria and presence of balloon cells. Nuclear changes showed increase in size and striking anisonucleosis, especially in CCl(4) and paracetamol treated mouse hepatocytes. Condensation of chromatin, nucleoli were fragmented and dispersed in beta-galactosamine induced hepatotoxic mice. These changes are remarkably striking in contrast to control animals. Treatment with CI-1, the herbal protein isolated from Cajanus indicus inhibited the pathogenesis of a majority of lesions produced by the hepatotoxins. Slender mitochondria, array of granular ER, presence of binucleated cells are the salient features of CI-1 treated hepatotoxic mice. Ultrastructurally, the hepatocytes of CI-1 treated mice were near normal. Thus, the herbal protein CI-1, may be a useful approach in the treatment of liver disorders for its potential in clinical medicine. PMID- 11483373 TI - Ethno-veterinary medicine: screening of Nigerian medicinal plants for trypanocidal properties. AB - Trypanosoma congolense and T. brucei bloodstream form parasites were propagated axenically in suitable standard media at 34 degrees C. The effects of 33 plant extracts, fractions and pure compounds were evaluated on two clones of T. brucei and drug-sensitive and multi-drug-resistant clones of T. congolense. The cytotoxic activity of the trypanocidal extracts was also evaluated on calf aorta endothelial cells in vitro. Of the extracts tested, 22% killed T. congolense IL 1180 at a concentration of 100 microg/ml while 18% killed 90-100% of T. brucei ILTat 1.4 at the same concentration. However, 6% of the active extracts killed 93% of a dyskinetoplastid form of T. brucei IL Tat 1.1, indicating that the intact kinetoplast is a target of some of the compounds tested. Of the 12 extracts that displayed activity against drug sensitive trypanosomes, 66.7% had trypanocidal activity on a multi-drug-resistant clone, T. congolense IL 3338. The extracts of Eugenia uniflora, Acacia artaxacantha, Terminalia ivorensis, T. superba and Alchornea cordifolia had median lethal concentrations of between 13 and 69 microg/ml on both the drug-sensitive, IL 1180 and multi-drug-resistant clone, IL 3338. The median lethal doses of the active plant extracts on the calf aorta endothelial cells varied between 112 and 13750 microg/ml while the calculated selective indices ranged between 0.71 and 246.8 indicating bright prospects for the development of some of these extracts as potential trypanocidal agents. PMID- 11483374 TI - Assessment of mutagenicity in Parthenium hysterophorus L. AB - The mutagenic potential of a crude extract of Parthenium hysterophorus L. was assessed in the Salmonella/microsome (Ames) assay and the mouse bone marrow micronucleus test. Results in the bacterial mutagenicity assay were negative for the five strains employed, e.g. TA 1535, TA1537, TA 98, TA 100 and TA 102, while cytotoxicity was evident in all cases at 5000 microg per plate, the highest concentration assayed. A decrease in toxicity was observed with exogenous mammalian metabolic activation (S9) or glutathione (5 micromol per plate). When mutagenicity was monitored after column chromatography fractionation of the crude, fraction 1 was mutagenic in strain TA 98 (+S9). Besides, cytotoxicity was found in fraction 5, where parthenin was eluted. The micronucleus test was negative in mice upon oral administration, at doses up to 96 mg of crude per kg. Bone marrow toxicity was not observed. The crude extract exhibited some in vitro pro-oxidant activity. It also inhibited lipid peroxidation (IC(50)=4.1 microg/ml) but failed to act as .OH scavenger. PMID- 11483375 TI - The in vitro effect of Helichrysi flos on microsomal lipid peroxidation. AB - Helichrysum arenarium (L.) Moench has long been known as a medicinal plant in Europe for its cholagogue, choleretic, hepatoprotective and detoxifying activities. Antioxidant properties of its main phenolics, flavonoids may be supposed to be responsible for these effects. The aim of this study was to verify the antioxidant properties of lyophilized water extracts with different polyphenol and flavonoid contents from inflorescences. The effects of natural extracts on microsomal fraction of rat liver were examined. Enzymatically induced lipid peroxidation and NADPH cytochrome c reductase activity in liver microsomes were measured by spectrophotometric methods. Results were compared with the activity of silibinin flavonoid, the main agent of well-known milk thistle (Silybum marianum L.). The natural plant extracts diminish the enzymatically induced lipid peroxidation in a concentration-dependent manner and reduce the cytochrome c dose dependently. The sample with higher polyphenol and flavonoid contents showed more stimulation of NADPH cytochrome c reductase. The lyophilized Helichrysi flos extracts proved to be more effective compared to silibinin in examined concentrations. PMID- 11483376 TI - Antimicrobial activity of Argentine plants used in the treatment of infectious diseases. Isolation of active compounds from Sebastiania brasiliensis. AB - Different extracts of Sebastiania brasiliensis, Sebastiania klotszchiana, Polygonum punctatum, Lithraea molleoides and Myrcianthes cisplatensis, all plants growing in Entre Rios Province and traditionally used as antiseptics, were tested against a set of Gram positive and Gram negative bacteria and fungi. All the species, with the exception of M. cisplatensis, presented activity against some of the microorganisms tested. A 50% hydroalcoholic extract of S. brasiliensis was selected for bioguided fractionation. Two antimicrobial compounds identified as methylgallate (MIC 128 microg/ml) and protocatechuic acid (MIC 128 microg/ml) were isolated apart from quercetin, kaempferol, quercitrin and gallic acid. PMID- 11483377 TI - Antiulcerogenic and analgesic effects of Maytenus aquifolium, Sorocea bomplandii and Zolernia ilicifolia. AB - Maytenus aquifolium (Celastraceae), Sorocea bomplandii (Moraceae) and Zolernia ilicifolia (Fabaceae) are native plants from the Tropical Atlantic Forest (Mata Atlantica, Brazil) known as "espinheira-santa". These plants are traditionally used as analgesic and antiulcerogenic medicine, with the same traditional uses of the true "espinheira-santa" (Maytenus ilicifolia, Celastraceae), an efficient antiulcerogenic agent. Pharmacological and toxicological studies with these plants have not been carried out. The purpose in this study was to evaluate the efficacy (analgesic and antiulcerogenic activities), safety (acute toxicity) and quality (phytochemical profile) of these three plants. The analgesic effect was analyzed by writhing and tail flick tests, while antiulcerogenic effect was performed through ulcer induction by ethanol and indomethacin/bethanecol assays. LD(50) and acute toxic effects, as well as phytochemical profiles of all plants also were carried. Surprisingly, the three plants showed analgesic and antiulcerogenic effects at dose of 1000 mg/kg, v.o. Maytenus aquifolium lowering all ulcerogenic parameters (ethanol test), but increased the ulcerogenic effects in the indomethacin/bethanecol test. Sorocea bomplandii produced antiulcerogenic effects in both experimental models used, while Zolernia ilicifolia showed significant effects only in indomethacin/bethanecol-induced gastric lesions. Pre treatment with Zolernia ilicifolia induced someone toxic effects. A phytochemical profile for each plant species was determined and its main chemical classes of compounds were described. PMID- 11483378 TI - Antimicrobial activity of Alstonia macrophylla: a folklore of bay islands. AB - The methanolic crude and methanol-aqueous extract of Alstonia macrophylla leaves and n-butanol part of the crude extract showed antimicrobial activity against various strains of Staphylococcus aureus, Staphylococcus saprophyticus, Streptococcus faecalis, Escherichia coli, Proteus mirabilis, Trichophyton rubrum, Trichophyton mentagrophytes var. mentagrophytes and Microsporum gypseum. The minimum inhibitory concentration (MIC) values ranges from 64 to 1000 microg/ml for bacteria and 32-128 mg/ml for dermatophytes. However, the strains of Pseudomonas aeruginosa, Klebsiella sp. and Vibrio cholerae showed resistance against in vitro treatment of the extracts up to 2000 microg/ml concentration, while the two yeast species were resistant even at 128 mg/ml concentration. The stem bark extract prepared similarly was found to be less active compared to the leaves. Phytochemical study indicates that the crude extract contains tannins, flavonoids, saponins, sterols, triterpene and reducing sugars. Further fractionation and purification of n-butanol part of the extract showed the presence of beta-sitosterol, ursolic acid, beta-sitosterol glucoside and a mixture of minor compounds only detected in TLC. PMID- 11483379 TI - Studies on pharmaceutical ethnobotany in the region of Pallars (Pyrenees, Catalonia, Iberian Peninsula). Part I. General results and new or very rare medicinal plants. AB - An ethnobotanical survey was carried out in the region called Pallars- constituted by two districts, Pallars Sobira and Pallars Jussa--situated in the Central Pyrenees, in North West Catalonia (Iberian Peninsula), with an approximate area of 2530 km(2) and a population of 19000. We obtained data on 437 plant species used for health care through interviews with 264 people. We detected 867 unreported or uncommon uses corresponding to 272 plant species, 52 of which had never or very rarely been cited as medicinal. To present the most important findings concerning the ethnopharmacology of the area studied, this first part includes the general results and the new or very scarcely reported medicinal vascular plants. Further papers will deal with unreported or very uncommon uses for known medicinal plant species and with medicinal non-vascular plants. PMID- 11483380 TI - Effects of Ferula gummosa Boiss. fractions on morphine dependence in mice. AB - Activity-guided fractionation of methanol-chloroform (1:1) extract of Ferula gummosa was carried out to investigate the isolation of the active component(s) responsible for the alleviation of morphine withdrawal syndrome induced by naloxone. Dependence was induced using subcutaneous (s.c.) injections of morphine daily for three days. On day 4, morphine was injected 0.5 h before the interaperitoneal (i.p.) injections of fractions or diazepam (5 mg/kg, i.p.) as positive control. Naloxone was injected (5 mg/kg, s.c.) 2 h after the final dose of morphine. The number of episodes of jumping during 30 min after the injection of naloxone was considered as the intensity of the withdrawal syndrome. The methanol-chloroform (1:1) extract of the aerial parts of plant was prepared and partitioned between water and chloroform. The active chloroform layer was concentrated and partitioned between methanol-water (9:1) and n-hexane. Activity was observed in the hydroalcoholic layer. This layer was concentrated and partitioned further between methanol-water (3:2) and chloroform. The chloroform layer showed a dose-dependent and significant activity. For all fractions the activity was observed at 470 mg/kg. Further purification on silica gel column chromatography gave a pure compound, which was 10 times as effective as the crude extract. The results of this study indicated that the plant extract contained component(s) that could be useful for the alleviation of morphine withdrawal syndrome. PMID- 11483381 TI - Biological screening of plants of the Venezuelan Amazons. AB - A total of 21 extracts derived from 17 different plant species collected in Venezuelan Amazons have been tested for the following biological activities: cardiovascular activity, brine shrimp lethality, and inhibitory effects on the hydrolysis of glucose-6-phosphate in intact and disrupted microsomes. Eight extracts diminished rat blood pressure with or without changes in heart rate. The fruit extract of Swartzia leptopetala and the leaf and twig extract of Connarus lambertii resulted in death of experimental animals. The majority of extracts (17 extracts) showed significant toxicity against Artemia salina. Concerning the hydrolysis of glucose-6-phosphate, better inhibitory effects were observed in intact microsomes than in disrupted ones for all the extracts, suggesting that these extracts intervene with variable potency in glucose-6-phosphate transport through the microsomal membrane. PMID- 11483382 TI - Immunomodulatory effect of concentrated lime juice extract on activated human mononuclear cells. AB - In this study, the in vitro immunomodulatory effect of concentrated juice of Citrus aurantifolia cv. swingle (Lime) was investigated. Clarified fresh lime juice was concentrated by freeze-drying. After dialysis against phosphate buffered saline and sterilization by a Millipore filter, it was used for further experiments. Immunogenic property of the CLJ extract was documented by production of specific polyclonal antibodies in rabbits. The immunomodulatory effect of the extract was tested in mitogen activated cultured mononuclear cells. The culture results indicated that proliferation of phytohemagglutinin (PHA) activated mononuclear cells were significantly inhibited by 250 and 500 microg/ml of CLJ extract, whereas only 500 microg/ml of the extract could inhibit proliferation of staphylococcal protein A (SPA) activated mononuclear cells (P<0.05). The abrogation of this inhibitory effect of the CLJ extract was noted by adding anti CLJ antibody to the lymphocyte culture. Considering these data, it can be concluded that the CLJ extract possesses immunomodulatory principles, which may mainly be due to the protein components of the extract. PMID- 11483383 TI - A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part V. Evaluation of the antimalarial activity of plants used by the Tacana Indians. AB - One hundred and twenty-five extracts of 122 different plant species traditionally used by the Tacana, a native community living in lowland forest at the base of the last foothills of the Cordillera Oriental of the Bolivian Andes, were screened for antimalarial activity in vitro on Plasmodium falciparum chloroquine resistant (D2) and sensitive strains (F32), and were evaluated in vivo on rodent malaria Plasmodium berghei. Five ethanolic stembark extracts showed marked activity either in vitro or in vivo, and only one of them, Bowdichia virgilioides being traditionally used against malaria, was active in vitro (IC50=1 microg/ml on both strains) and in vivo (51% at 100 mg/kg). Other active extracts were from Caesalpinia pluviosa bark displaying activity in vitro against chloroquine resistant strain (IC50 8.3 microg/ml), traditionally used against dysentery; two Lauraceae bark extracts, Nectandra aff. hihua and Licaria canella respectively used for construction purposes and against stomach ache, both displaying activity in vitro against P. falciparum sensible and resistant strains (IC50 around 4 microg/ml); finally, the bark of a strongly aromatic Burseraceae, Protium glabrescens exuding an anti-inflammatory and analgesic resin, was active in vivo only (61% at 100 mg/kg). Results are discussed in relation with Tacana traditional medicine. PMID- 11483384 TI - Hypoglycemic action of Cucurbita ficifolia on Type 2 diabetic patients with moderately high blood glucose levels. AB - Ten Type 2 diabetic patients with moderately hyperglycemia, received a raw extract of Cucurbita ficifolia or potable water in a single dose of 4 ml/Kg body weight, in two different sessions at least separated by 1 week. Blood glucose levels were followed in both parts of the study by a period of 5 h. The patients had stopped their pharmacologic medication 24 h prior to each part of the study. As expected, no significant changes on blood glucose were observed after giving the vehicle; however, the oral administration of C. ficifolia was followed by a significant decrease in blood glucose levels, from 12.07+/-1.69 mM (217.2+/-30.4 mg/dl) to 9.42+/-1.96 mM (169.6+/-35.3 mg/dl) 3 h after and to 8.37+/-1.74 mM (150.8+/-31.3 mg/dl) 5 h after the extract administration. The hypoglycemic action of Cucurbita ficifolia agrees with its effects previously observed in laboratory animals. The results of this preliminary study do not give indications for the mechanism of this hypoglycemic effect. PMID- 11483385 TI - Antimutagenicity and induction of anticarcinogenic phase II enzymes by basidiomycetes. AB - Extracts from Phellinus linteus, Phellinus igniarius, and Agrocybe cylindracea have been tested for their antimutagenic properties against direct-acting mutagens [4-nitro-o-phenylenediamine (NPD) and sodium azide (NaN(3))] and indirect-acting mutagens [2-aminofluorene (2-AF) and benzo[a]pyrene (B[a]P)], using the Salmonella typhimurium tester strains TA 98 and TA 100. In addition, the chemopreventive potentials of these extracts to induce NAD(P)H:quinone oxidoreductase (QR) and glutathione S-transferase (GST) activities and glutathione (GSH) level extracts from the filtrate of the cultured broth of P. linteus, polysaccharide extracts from the cultured broth (PI I) and mycelia (PI II) and water extract of fruiting bodies (PI II) of P. igniarius, and polysaccharide extracts from the cultured broth (AC I) and mycelia (AC II) of A. cylindracea showed inhibitory effects on the mutagenic activities induced by the direct-acting mutagens, NPD and NaN(3), and the indirect-acting mutagens, 2-AF and B[a]P. QR was induced with PI I, PI II, AC I, and AC II, and GST activity was induced with PL I, PL II, PI I, PI II, PI III and AC I in murine Hepa1c1c7 cell culture. In addition, PL I, PL II, PI I, PI II, PI III and AC II increased glutathione level. These results suggest that P. linteus, P. igniarius, and A. cylindracea have antimutagenic activities and may play a role in the prevention of cancer by inducing QR and GST activities and increasing GSH level. PMID- 11483386 TI - Anti-bacterial activity of Galega officinalis L. (Goat's Rue). AB - Alcoholic extracts of Goat's Rue (Galega officinalis L.; Papilionaceae) were tested on Gram +ve and Gram -ve bacteria as the plant was claimed to hasten skin healing after surgery. Ethanolic (60%) extract exhibited significant inhibition on growth of both Gram +ve and Gram -ve bacteria. PMID- 11483387 TI - Anti-HIV activity of medicinal plant extracts. AB - As part of our screening of anti-AIDS agents from natural sources, ethanolic and aqueous extracts of 15 medicinal plants widely used in the folk medicine of the Iberian Peninsula were evaluated in vitro. Most of the extracts tested were relatively nontoxic to human lymphocytic MT-2 cells, but only the extracts of Tuberaria lignosa and Sanguisorba minor magnolii exhibited anti-HIV activity in an in vitro MTT assay. The aqueous extracts of these plants showed inhibitory effects against HIV-1 induced infections in MT-2 cells at concentrations ranging from 12.5 to 50 microg/ml and 50 microg/ml, respectively. Both extracts showed no appreciable cytotoxicity at these concentrations. PMID- 11483388 TI - Effects of Inula viscosa leaf extracts on abortion and implantation in rats. AB - The anti-implantation and mid-term abortifacient effects exhibited by administration of Inula viscosa leaf extracts, were studied in rats. The aqueous extract administered i.p. on day 1-6 of gestation, totally diminished fetal implantation and caused a significant (P<0.05) reduction in the number of corpora lutea and blood progesterone levels. Meanwhile, administration on day 13-15 of gestation exhibited mid-term abortion. Furthermore, petroleum ether and dichloromethane, but not methanol, extracts exhibited pronounced abortifacient effects. The results of this study are in agreement with the traditional reputation of this plant as abortifacient. Further, the plant possesses anti implantational and luteolytic effects. PMID- 11483389 TI - Antibacterial activity of leaf essential oils and their constituents from Cinnamomum osmophloeum. AB - The antibacterial activities of the essential oils from leaves of two Cinnamomum osmophloeum clones (A and B) and their chemical constituents were investigated in this study. The nine strains of bacteria, including Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus (MRSA), Klebsiella pneumoniae, Salmonella sp., and Vibrio parahemolyticus, were used in the antibacterial tests. Results from the antibacterial tests demonstrated that the indigenous cinnamon B leaf essential oils had an excellent inhibitory effect. The MICs (minimum inhibitory concentrations) of the B leaf oil were 500 microg/ml against both K. pneumoniae and Salmonella sp. and 250 microg/ml against the other seven strains of bacteria. Cinnamaldehyde possessed the strongest antibacterial activity compared to the other constituents of the essential oils. The MICs of cinnamaldehyde against the E. coli, P. aeruginosa, E. faecalis, S. aureus, S. epidermidis, MRSA, K. pneumoniae, Salmonella sp., and V. parahemolyticus were 500, 1000, 250, 250, 250, 250, 1000, 500, and 250 microg/ml, respectively. These results suggest that C. osmophloeum leaf essential oil and cinnamaldehyde are beneficial to human health, having the potential to be used for medical purposes and to be utilized as anti-bacterial additives in making paper products. PMID- 11483390 TI - The cytotoxicity and chemical constituents of the hexane fraction of Typhonium flagelliforme (Araceace). AB - The plant, Typhonium flagelliforme (Araceae), commonly known as the "rodent tuber" in Malaysia, is often used as an essential ingredient of herbal remedies for alternative cancer therapies. The hexane extract of this plant was evaluated for cytotoxic activity against in vitro culture on P388 murine leukaemia cells and showed weak IC(50) of 15 microg/ml. The partial chemical constituents were identified as methyl esters of hexadecanoic acid, octadecanoic acid, 9 octadecenoic acid and 9,12-octadecadienoic acid. In addition, several common aliphatics were identified as dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane and eicosane. The unique methyl ester of 13-phenyltridecanoic acid was isolated and positively identified using spectroscopic methods. None of the identified compounds showed or are known to have cytotoxic behaviour. PMID- 11483391 TI - Mortality in pediatric epilepsy. AB - Mortality in pediatric epilepsy is the subject of this review. Epilepsy in both adults and children increases the risk of premature death. Conditions that are comorbid with epilepsy may carry an increased mortality risk. Patients with neurologic compromise may be at risk for aspiration and critical respiratory disease. Epilepsy per se enhances the risks of accidents, particularly drowning. Epilepsy may result in unwitnessed or, less frequently, witnessed sudden death. Witnessed sudden death frequently involves an observed seizure. Cardiac and respiratory mechanisms for epileptic sudden death have been proposed. Reducing the number of seizures should reduce the risk for sudden death. Awareness of the increased risk for premature death associated with epilepsy may enhance patients' and parents' cooperation with therapy. PMID- 11483392 TI - Lafora's disease: towards a clinical, pathologic, and molecular synthesis. AB - Lafora's disease is one of five inherited progressive myoclonus epilepsy syndromes. It is an autosomal-recessive disorder with onset in late childhood or adolescence. Characteristic seizures include myoclonic and occipital lobe seizures with visual hallucinations, scotomata, and photoconvulsions. The course of the disease consists of worsening seizures and an inexorable decline in mental and other neurologic functions that result in dementia and death within 10 years of onset. Pathology reveals pathognomonic polyglucosan inclusions that are not seen in any other progressive myoclonus epilepsy. Lafora's disease is one of several neurologic conditions associated with brain polyglucosan bodies. Why Lafora's polyglucosan bodies alone are associated with epilepsy is unknown and is discussed in this article. Up to 80% of patients with Lafora's disease have mutations in the EPM2A gene. Although common mutations are rare, simple genetic tests to identify most mutations have been established. At least one other still unknown gene causes Lafora's disease. The EPM2A gene codes for the protein laforin, which localizes at the plasma membrane and the rough endoplasmic reticulum and functions as a dual-specificity phosphatase. Work toward establishing the connection between laforin and Lafora's disease polyglucosans is underway, as are attempts to replace it into the central nervous system of patients with Lafora's disease. PMID- 11483393 TI - Diagnostic profile of neonatal hypotonia: an 11-year study. AB - The profile of disorders presenting with neonatal hypotonia to the neonatal intensive care unit has not been studied previously. An 11-year retrospective cohort study of neonates, who were identified through computer database records and were admitted to the Neonatal Intensive Care Unit from January 1989 to December 1999 at the Montreal Children's Hospital (Montreal, Quebec), is presented. The final diagnoses, tests obtained, and outcome were determined from a structured review of the subject's hospital record. The database search generated 95 records, of which 50 neonates met the inclusion criteria. The hypotonia was classified as central in 33 patients (66%) and peripheral in 17 (34%). Hypoxic-ischemic encephalopathy (n = 13), Prader-Willi syndrome (n = 6), myotonic dystrophy (n = 6), other muscle disorders (n = 6), chromosomal disorders (n = 4), and peripheral nerve disorders (n = 3) were the most common diagnoses. The genetic tests of highest yield were fluorescent in situ hybridization for Prader-Willi syndrome, DNA methylation studies for Prader-Willi syndrome, trinucleotide repeat testing for myotonic dystrophy, and karyotype analysis. A diagnostic approach is proposed based on the results. PMID- 11483394 TI - Incidence of epileptic discharge in various epileptic syndromes. AB - Three hundred eight patients with childhood and adolescent epilepsy were examined to clarify the incidence of epileptic discharges on initial and follow-up electroencephalograms. Epileptic discharges were found in 75.6% patients on the initial electroencephalogram, which is higher than figures previously reported for adults. The cumulative incidence of epileptic discharges was 92.3% by the third electroencephalogram recording. However, in 17.1% patients with nonspecific idiopathic generalized epilepsy, no epileptic discharges were found even after three electroencephalogram recordings. The incidence of epileptic discharges in patients with generalized epilepsy (84.3%) was significantly higher than in patients with localization-related epilepsy (71.6%). The incidence of epileptic discharges in patients with partial seizures was lower than those in patients with generalized seizures. The incidence of epileptic discharges was low in the 0 to 3-year-old and 15- to 20-year-old groups, and high in the 3- to 12-year-old groups. In the positive epileptic discharge patients, 38.8% of electroencephalograms were abnormal only during the waking or sleeping portion of the recordings. Knowing the incidence of epileptic discharges for each type of epilepsy will be useful in planning further electroencephalogram research and performing electroencephalograms in the clinical setting. PMID- 11483395 TI - Thyroid hormones in epileptic children receiving carbamazepine and valproic acid. AB - Carbamazepine and valproic acid are effective antiepileptic drugs for treating many types of epilepsy. Although they are well tolerated, many effects on endocrine function have been reported. Changes in serum thyroid hormones levels in 37 children with epilepsy during carbamazepine and valproic acid therapy were analyzed, and the thyroid hormone concentration after thyrotropin-releasing hormone test was evaluated. Serum thyroxine and free thyroxine levels were significantly lower in patients treated with carbamazepine and carbamazepine plus valproic acid than in the control subjects; serum thyroxine and free thyroxine concentrations were unaffected by valproic acid monotherapy. Serum triiodothyronine and free triiodothyronine concentrations were similar in the three groups of patients studied. Thyroid-stimulating hormone serum levels were normal in all patients, and the thyrotropin responses to the thyrotropin releasing hormone were similar to control group. Our data suggest that children treated with carbamazepine may have subclinical signs of hypothyroidism, and these changes are more evident if carbamazepine is given in association with valproic acid, while no alteration in thyroid hormones can be found with valproic acid monotherapy. Thyroid-stimulating hormone and thyrotropin-releasing hormone levels do not seem to be affected by these drugs, suggesting that hypothalamic function is not affected in these children. PMID- 11483396 TI - Traditional Chinese medicine (tongue acupuncture) in children with drooling problems. AB - Tongue acupuncture is an innovative technique in traditional Chinese medicine. We have demonstrated that specific tongue acupoints are related to various functional domains. This study aimed to assess the efficacy of tongue acupuncture in children with neurologic disability who had severe drooling problems. We conducted an intent-to-treat study in a cohort of 10 children. A continuous course of tongue acupuncture was performed daily to five acupoints in the tongue for a total of 30 sessions. Standardized outcome measures of drooling were evaluated by a blinded assessor to study the efficacy at baseline and after a course of treatment. Statistically significant improvement was noted in the following outcome measures: (1) mean visual analog scale (VAS) decreased from 6.6 (pre-TAC) to 4.67 (post-TAC) (P = 0.002); (2) mean drooling quotient (DQ) decreased from 14.016% (pre-TAC) to 8.335% (post-TAC) (P = 0.0078); and (3) mean drooling score (DS) decreased from 7.4 (pre-TAC) to 4.4 (post-TAC) (P = 0.002). This study demonstrated the efficacy of tongue acupuncture as an adjunctive or alternative treatment for patients with drooling problems and can be integrated as part of the oromotor stimulation program, drooling program, and behavioral modification program before subjecting the patient to invasive surgical procedures on the salivary glands. PMID- 11483397 TI - Age-dependent effects of trihexyphenidyl in extrapyramidal cerebral palsy. AB - Trihexyphenidyl (Artane) is a centrally active muscarinic antagonist commonly used to treat patients with generalized dystonia. In a retrospective survey of 22 consecutive children with extrapyramidal cerebral palsy, we evaluated trihexyphenidyl on upper extremity and lower extremity function, expressive language, and drooling. Functional changes were assessed using a parental questionnaire (rating scale 1-5: from 1 = little or no change to 5 = tremendous change, with scores in either a positive or negative direction). Improvements of +4 or +5 were reported in eight children for upper extremity function, in eight children for verbal expressive language, in five for drooling, and in none for lower extremity function. Using bivariate linear regression modeling to investigate variables associated with treatment effects, there was a significant inverse relationship between age at initiation of medication and therapeutic response. Furthermore, beneficial responses were specific to upper-extremity function and expressive language. These results suggest that younger children are more likely to respond to trihexyphenidyl and that primary functional benefits include improved fine motor abilities and expressive language. A prospective masked study with a standardized clinical instrument is needed to confirm these findings. PMID- 11483398 TI - Late-onset GM2 gangliosidosis presenting as burning dysesthesias. AB - Two brothers with a painful neuropathy as a component of late-onset GM2 gangliosidosis of the Sandhoff type are presented. A dramatic response of the severe dysesthesias to amitriptyline and gabapentin is described. Symptomatic sensory neuropathy may be a component of late-onset GM2 gangliosidosis. PMID- 11483399 TI - Congenital insensitivity to pain with anhidrosis. AB - Congenital insensitivity to pain with anhidrosis is an autosomal-recessive disorder resulting from defective neural crest differentiation with loss of the first-order afferent system, which is responsible for pain and temperature sensation. There is also a neuronal loss in the sympathetic ganglia. Lack of sweating, hyperthermia, and infections of bones are main features of the disorder; however, contradictory results have been published regarding eccrine sweat gland innervation. A 5-year-old male patient with typical clinical manifestations of congenital insensitivity to pain with anhidrosis is presented. Immunohistochemistry with antibodies against S100 protein and neuron-specific enolase failed to reveal nerve fibers in the vicinity of the eccrine sweat glands. The roles of the nerve growth factor and tyrosine kinase receptor gene mutations in the pathogenesis of the disease are also discussed. PMID- 11483400 TI - Mycoplasma pneumoniae infection, meningoencephalitis, and hemophagocytosis. AB - Central nervous system manifestations are a common extrapulmonary complication of Mycoplasma pneumoniae infection, of which encephalitis is a well-recognized abnormality in children. In this report the first description of M. pneumoniae infection simultaneously complicated by meningoencephalitis and hemophagocytosis is presented. PMID- 11483401 TI - Three patients with ophthalmoplegia associated with Campylobacter jejuni. AB - Cranial polyneuropathy is idiopathic in most patients. Idiopathic cranial polyneuropathy is an acute postinfectious syndrome, along with Guillain-Barre syndrome and Miller Fisher syndrome, in which the common preceding pathogen is Campylobacter jejuni. Serum anti-GQ1b antibodies are elevated in Miller Fisher syndrome and Guillain-Barre syndrome with ophthalmoplegia. Three patients with idiopathic cranial polyneuropathy with predominant ocular involvement are presented. C. jejuni isolated from stool specimens belonged to Penner serotypes O:4, O:23, and O:33. Serum anti-GQ1b antibodies were elevated in all patients but demonstrated rapid reduction concomitant with clinical recovery. All patients recovered completely. Because both preceding C. jejuni infection and elevated anti-GQ1b antibodies decreasing with time were seen in all patients, the pathogenesis of idiopathic cranial polyneuropathy with ophthalmoplegia may be similar to that of Miller Fisher syndrome. PMID- 11483402 TI - Moyamoya syndrome with spherocytosis: effect of splenectomy on strokes. AB - A 3-year-old male patient with hereditary spherocytosis who developed moyamoya syndrome, presenting hemiplegia, and slurred speech is reported. Transient ischemic attacks occurred repeatedly with hemolytic crises. Magnetic resonance imaging and angiography revealed bilateral occlusion of the internal carotid and middle cerebral arteries with the formation of moyamoya vessels and multiple infarctions in the basal ganglia. Although splenectomy can increase the risk of stroke, no stroke occurred after splenectomy. On aspirin and dipyridamole therapy the patient has been free of neurologic deficits and progression of the vasculopathy for 5 years. This rare observation suggests that anemic hypoxia more greatly contributes to the progression of moyamoya syndrome than postsplenectomy thrombocytosis or reduced deformability of spherocytes. PMID- 11483403 TI - Serial MRI findings in the Costa Rican variant of neuronal ceroid-lipofuscinosis. AB - Widespread cerebral atrophy and basal ganglia involvement are highly suggestive imaging features of the variants of late infantile type neuronal ceroid lipofuscinosis. In the presence of clinical findings indicative of neuronal ceroid-lipofuscinosis, neuroimaging procedures are highly recommended to differentiate the variants from classic late infantile neuronal ceroid lipofuscinosis. The clinical features and follow-up magnetic resonance imaging studies in a patient with the Costa Rican variant of late infantile neuronal ceroid-lipofuscinosis is presented. These procedures were of the utmost importance to observe the progression of the neurologic ailment and the extent of the cerebral and cerebellar abnormalities. PMID- 11483404 TI - Involuntary movements in infantile cobalamin deficiency appearing after treatment. AB - Involuntary movements may be a symptom in most infants who present with neurologic syndrome of infantile cobalamin (vitamin B12) deficiency. In this report, two infants with cobalamin deficiency are presented. These patients also developed a striking movement disorder that appeared a few days after treatment with intramuscular cobalamin. The movement disorder was characterized by severe involuntary movements, which were a combination of tremor and myoclonus particularly involving tongue, face, pharynx, and legs. The neurologic symptoms improved within a few days after the administration of clonazepam. In each patient the mother was also cobalamin deficient and the infant was solely breast fed. The cause of involuntary movements that can appear rarely after treatment in infantile cobalamin deficiency is not known. Besides initial neurologic presenting symptoms of cobalamin deficiency, the occurrence of involuntary movements after treatment should also receive attention. This movement disorder may disappear spontaneously, or an additional treatment may be an alternative approach if the symptoms are severe. PMID- 11483405 TI - Transcription factors p53 and HIF-1alpha as targets of nitric oxide. AB - It is widely recognized that the production of nitric oxide (NO) from L-arginine metabolism is an essential determinate of diverse signalling cascades throughout the body, with a major impact during nonspecific host defence. Biological actions of NO and derived species comprise physiological as well as pathological entities, with an impressive and steadily growing number of signalling pathways and/or protein targets being involved. It is now appreciated that NO not only acts as an effector molecule but also as an autocrine as well as paracrine modulator of rapid and delayed cellular responses. Among multiple targets the tumour suppressor p53 and the hypoxia inducible factor-1alpha (HIF-1alpha) emerged. Accumulation of p53 in response to NO delivery may account for an interference in cell cycle progression and/or initiation of apoptosis that is found in close correlation with inducible NO synthase (NOS) expression. Quite similarly, accumulation of HIF-1alpha not only occurs during hypoxia, but also under conditions of NO delivery, thus mimicking a situation of reduced oxygen availability. Interestingly, p53 and HIF-1alpha share regulatory elements that cause protein stabilization in part as a result of impaired ubiquitin-evoked protein degradation. Here, we summarize current knowledge on the impact of NO on p53- and HIF-1alpha-stabilization and we will discuss pathophysiological consequences. These examples may help to shape and refine current concepts of NO action with an emphasis on transcription factor regulation. PMID- 11483406 TI - Mechanism of prostaglandin D(2)-stimulated heat shock protein 27 induction in osteoblasts. AB - We previously showed that prostaglandin D(2) (PGD(2)) stimulates activation of protein kinase C (PKC). We investigated whether PGD(2) stimulates the induction of heat shock protein (HSP) 27 and HSP70 in osteoblast-like MC3T3-E1 cells and the mechanism underlying the induction. PGD(2) increased the levels of HSP27 while having little effect on HSP70 levels. PGD(2) stimulated the accumulation of HSP27 dose dependently in the range between 10 nM and 10 microM. PGD(2) induced an increase in the levels of mRNA for HSP27. The PGD(2)-stimulated accumulation of HSP27 was reduced by staurosporine or calphostin C, inhibitors of PKC. PGD(2) induced the phosphorylation of p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase. The HSP27 accumulation induced by PGD(2) was significantly suppressed by PD98059, an inhibitor of the upstream kinase of p44/p42 MAP kinase, or SB203580, an inhibitor of p38 MAP kinase. Calphostin C suppressed the PGD(2) induced phosphorylation of p44/p42 MAP kinase and p38 MAP kinase. PD98059 or SB203580 suppressed the PGD(2)-increased levels of mRNA for HSP27. These results strongly suggest that PGD(2) stimulates HSP27 induction through p44/p42 MAP kinase activation and p38 MAP kinase activation in osteoblasts and that PKC acts at a point upstream from both the MAP kinases. PMID- 11483407 TI - Tumor necrosis factor alpha-induced activation of downstream NF-kappaB site of the promoter mediates epithelial ICAM-1 expression and monocyte adhesion. Involvement of PKCalpha, tyrosine kinase, and IKK2, but not MAPKs, pathway. AB - TNF-alpha induced an increase in intercellular adhesion molecule-1 (ICAM-1) expression in human A549 epithelial cells and immunofluorescence staining confirmed this result. The enhanced ICAM-1 expression was shown to increase the adhesion of U937 cells to A549 cells. Tyrosine kinase inhibitors (genistein or tyrphostin 23) or phosphatidylcholine-specific phospholipase C (PC-PLC) inhibitor (D 609) attenuated TNF-alpha-induced ICAM-1 expression. TNF-alpha produced an increase in protein kinase C (PKC) activity and this effect was inhibited by D 609. PKC inhibitors (staurosporine, Ro 31-8220, calphostin C, or Go 6976) also inhibited TNF-alpha-induced response. 12-O-Tetradecanoylphorbol-13-acetate (TPA), a PKC activator, stimulated ICAM-1 expression, this effect was inhibited by genistein or tyrphostin 23. Treatment of cells with TNF-alpha resulted in stimulation of p44/42 MAPK, p38, and JNK. However, TNF-alpha-induced ICAM-1 expression was not affected by either MEK inhibitor, PD 98059, or p38 inhibitor, SB 203580. A cell-permeable ceramide analog, C(2) ceramide, also stimulated the activation of these three MAPKs, but had no effect on ICAM-1 expression. NF kappaB DNA-protein binding and ICAM-1 promoter activity were enhanced by TNF alpha and these effects were inhibited by D 609, calphostin C, or tyrphostin 23, but not by PD 98059 or SB 203580. TPA also stimulated NF-kappaB DNA-protein binding and ICAM-1 promoter activity, these effects being inhibited by genistein or tyrphostin 23. TNF-alpha- or TPA-induced ICAM-1 promoter activity was inhibited by dominant negative PKCalpha or IKK2, but not IKK1 mutant. IKK activity was stimulated by both TNF-alpha and TPA, and these effects were inhibited by Ro 31-8220 or tyrphostin 23. These data suggest that, in A549 cells, TNF-alpha activates PC-PLC to induce activation of PKCalpha and protein tyrosine kinase, resulting in the stimulation of IKK2, and NF-kappaB in the ICAM-1 promoter, then initiation of ICAM-1 expression and neutrophil adhesion. However, activation of p44/42 MAPK, p38, and JNK is not involved in this event. PMID- 11483408 TI - Uncoupling of ATP-induced inositol phosphate formation and Ca(2+) mobilization by phorbol ester in canine cultured tracheal epithelial cells. AB - The regulation of the increase in inositol phosphates (IPs) production and intracellular Ca(2+) concentration ([Ca(2+)](i)) by protein kinase C (PKC) was investigated in canine cultured tracheal epithelial cells (TECs). Pretreatment of TECs with phorbol 12-myristate 13-acetate (PMA, 1 microM) for 30 min attenuated the ATP- and UTP-induced IPs formation and Ca(2+) mobilization. The concentrations of PMA that gave half-maximal (EC(50)) inhibition of ATP- and UTP induced IPs accumulation and an increase in [Ca(2+)](i) were 5-10 and 4-12 nM, respectively. Prior treatment of TECs with staurosporine (1 microM), a PKC inhibitor, partially inhibited the ability of PMA to attenuate ATP- and UTP induced responses, suggesting that the inhibitory effect of PMA is mediated through the activation of PKC. Furthermore, analysis of cell extracts by Western blotting with antibodies against different PKC isozymes revealed that TECs expressed PKC-alpha, -betaI, -betaII, -gamma, -delta, -epsilon, -theta, and zeta. With PMA treatment of the cells for various times, translocation of PKC alpha, -betaI, -betaII, -gamma, -delta, -epsilon, and -theta from the cytosol to the membrane was seen after 5- and 30-min and 2- and 4-h treatment. However, 6-h treatment caused a partial down-regulation of these PKC isozymes. PKC-zeta was not significantly translocated and down-regulated at any of the times tested. In conclusion, these results suggest that activation of PKC may inhibit the phosphoinositide (PI) hydrolysis and consequently attenuate the [Ca(2+)](i) increase or inhibit independently both responses to ATP and UTP. The translocation of PKC-alpha, -betaI, -betaII, -delta, -epsilon, -gamma, and -theta induced by PMA caused an attenuation of ATP- and UTP-induced IPs accumulation and Ca(2+) mobilization in TECs. PMID- 11483409 TI - Mechanisms of bradykinin-mediated Ca(2+) signalling in canine cultured corneal epithelial cells. AB - Experiments were designed to differentiate the mechanisms of bradykinin receptors mediating the changes in intracellular Ca(2+) concentration ([Ca(2+)](i)) in canine cultured corneal epithelial cells (CECs). Bradykinin and Lys-bradykinin caused an initial transient peak of [Ca(2+)](i) in a concentration-dependent manner, with half-maximal stimulation (pEC(50)) obtained at 6.9 and 7.1, respectively. Pretreatment of CECs with pertussis toxin (PTX) or cholera toxin (CTX) for 24 h did not affect the bradykinin-induced [Ca(2+)](i) changes. Application of Ca(2+) channel blockers, diltiazem and Ni(2+), inhibited the bradykinin-induced Ca(2+) mobilization, indicating that Ca(2+) influx was required for the bradykinin-induced responses. Addition of thapsigargin (TG), which is known to deplete intracellular Ca(2+) stores, transiently increased [Ca(2+)](i) in Ca(2+)-free buffer, and subsequently induced Ca(2+) influx when Ca(2+) was readded to this buffer. Pretreatment of CECs with TG completely abolished bradykinin-induced initial transient [Ca(2+)](i), but had slight effect on bradykinin-induced Ca(2+) influx. Pretreatment of CECs with 1-[beta-[3-(4 methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole (SKF96365) and 1-(6 ((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122) inhibited the bradykinin-induced Ca(2+) release and Ca(2+) influx, consistent with the inhibition of receptor-gated Ca(2+) channels and phospholipase C (PLC) in CECs, respectively. These results demonstrate that bradykinin directly stimulates B(2) receptors and subsequently Ca(2+) mobilization via a PTX-insensitive G protein in canine CECs. These results suggest that bradykinin-induced Ca(2+) influx into the cells is not due to depletion of these Ca(2+) stores, as prior depletion of these pools by TG has no effect on the bradykinin-induced Ca(2+) influx that is dependent on extracellular Ca(2+) in CECs. PMID- 11483410 TI - Detection of calmodulin-binding proteins and calmodulin-dependent phosphorylation linked to calmodulin-dependent chemotaxis to folic and cAMP in Dictyostelium. AB - Calmodulin (CaM) antagonists, trifluoperazine (TFP) or calmidazolium (R24571), dose-dependently inhibited cAMP and folic acid (FA) chemotaxis in Dictyostelium. Developing, starved, and refed cells were compared to determine if certain CaM binding proteins (CaMBPs) and CaM-dependent phosphorylation events could be identified as potential downstream effectors. Recombinant CaM ([35S]VU-1-CaM) gel overlays coupled with cell fractionation revealed at least three dozen Ca(2+) dependent and around 12 Ca(2+)-independent CaMBPs in Dictyostelium. The CaMBPs associated with early development were also found in experimentally starved cells (cAMP chemotaxis), but were different for the CaMBP population linked to growth phase cells (FA chemotaxis). Probing Western blots with phosphoserine antibodies revealed several phosphoprotein bands that displayed increases when cAMP responsive cells were treated with TFP. In FA-responsive cells, several but distinct phosphoproteins decreased when treated with TFP. These data show that unique CaMBPs are present in growing, FA-chemosensitive cells vs. starved cAMP chemoresponsive cells that may be important for mediating CaM-dependent events during chemotaxis. PMID- 11483411 TI - Protein kinase C signalling pathway is involved in the regulation of vascular endothelial growth factor expression in human bladder transitional carcinoma cells. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic factor associated with the growth and metastasis of various cancers and plays a prominent role in vesical angiogenesis regulation. In this study, we investigated the effect of the phorbol 12-myristate 13-acetate (PMA) on the expression of VEGF in human bladder transitional carcinoma cells (RT4). RT4 cells expressed three VEGF isoforms (VEGF(189), VEGF(165), VEGF(121)). PMA increased VEGF mRNA expression time-dependently with a peak at 4 h. PMA increased the half-life of VEGF mRNA. The amount of VEGF protein in conditioned media was increased by PMA in a dose-dependent manner with a maximal effect at 10(-7) M. Staurosporine and calphostin C (PKC inhibitors) decreased PMA-induced VEGF mRNA expression as opposed to protein kinase A or cyclic nucleotide-dependent protein kinase inhibitors. Thus, in RT4 cells, VEGF expression is up-regulated by PMA via the PKC signalling pathway and according to a posttranscriptional mechanism. PMID- 11483412 TI - Dual regulation of sphingosine 1-phosphate-induced phospholipase D activity through RhoA and protein kinase C-alpha in C2C12 myoblasts. AB - Previous studies showed that in C2C12 cells, phospholipase D (PLD) and its known regulators, RhoA and protein kinase Calpha (PKCalpha), were downstream effectors in sphingosine 1-phosphate (SPP) signalling. Moreover, the role of PKC for SPP mediated PLD activation and the requirement of PKCalpha for RhoA translocation were reported. The present results demonstrated that inactivation of RhoA, by overexpression of RhoGDP dissociation inhibitor (RhoGDI) as well as treatment with C3 exotoxin, attenuated SPP-stimulated PLD activity, supporting the involvement of RhoA in the stimulation of PLD activity by the bioactive lipid in C2C12 myoblasts. In addition, the effect of PKCalpha inhibitor Go6976 on the SPP induced PLD activation in myoblasts, where RhoA function was inactivated, was consistent with a dual regulation of the enzyme through RhoA and PKCalpha. Interestingly, the subcellular distribution of PLD isoforms, RhoA and PKCalpha, in SPP-stimulated cells supported the view that the functional relationship between the two PLD regulators, demonstrated to occur in SPP signalling, represents a novel mechanism of regulation of specifically localized PLD. PMID- 11483413 TI - The diagnostic value of galactography in patients with nipple discharge. AB - To evaluate preoperative galactographic findings in the differentiation between the benign and malignant lesions in patients presenting spontaneous nipple discharge without mass. Of the 215 patients who have undergone the galactography, 181 cases with abnormal galactography had surgery performed. All galactrograms were reviewed and galactographic findings were correlated to the pathological results to determine diagnostic differentiation. Of the 181 cases we operated on, 112 cases were macroscopically bloody, with 30 cases having cancers (26.8%). Fifty-four cases with serous discharge had seven cancer cases (13.0%). No cancer cases with other color discharge were found. Of the 37 cancer cases, 11 cases had lesions located in the main mammary ducts (lactiferous duct and the segmental duct) (29.7%) and 26 cases had lesions in the peripheral ducts (the subsegmental duct and its branches) (70.3%) (P<.05). Of 113 cases with benign proliferative ductal lesions, 88 cases were located in the main mammary duct (77.9%) and 25 cases in the peripheral mammary duct (22.1%) (P<.05). Otherwise, 29 cancer cases (82.9%) had ductal obstructions and 28 cancer cases (75.7%) had irregular intraductal defects that appeared in the galactograms, which is different from the 113 benign proliferative ductal lesion cases that had 88 cases (71.7%) with ductal dilatation and 90 cases (79.6%) with lobular or smooth intraductal defects (P<.05). These results showed that the cancer cases had a higher rate of locating in the peripheral duct, irregular intraductal duct defects, and ductal obstruction, and a lower rate associated with ductal dilatation or torsion. The galactographic findings were evaluated using the tumor location, types of intraductal defects, ductal obstruction, and dilatation. Preoperative diagnostic galactography is useful in differentiating between the benign or malignant lesions in patients with spontaneous nipple discharge. PMID- 11483414 TI - Cerebral hemiatrophy caused by multiple developmental venous anomalies involving nearly the entire cerebral hemisphere. AB - The authors present a case of multiple developmental venous anomalies (DVAs) that involved nearly the entire cerebral hemisphere and that were diagnosed by magnetic resonance (MR) imaging and selective cerebral angiography. The ipsilateral cerebral hemisphere was mildly atrophic, and there were focal white matter lesions on the MR images. Angiography showed multiple DVAs involving nearly the entire right cerebral hemisphere. We suggest that longstanding venous congestion caused the atrophy and white matter lesions. PMID- 11483415 TI - The posterior inferior cerebellar artery arising from the extracranial segment of the internal carotid artery via the hypoglossal canal without an interposed segment of the basilar artery: a persistent primitive hypoglossal artery variant. AB - The authors present a rare case of a persistent primitive hypoglossal artery (PHA) variant ending in the posterior inferior cerebellar artery (PICA) without an interposed segment of the basilar artery and describe briefly the possible embryogenesis of this anomaly. PMID- 11483416 TI - Transcranial color Doppler sonography in the emergency diagnosis of middle cerebral artery occlusion in a patient after head injury. AB - The purpose of this report is to highlight the utility of transcranial color Doppler sonography (TCCDS) in the emergency diagnosis of an ischemic stroke in a 64-year-old patient after head injury. An emergency CT identified a subdural hematoma, subarachnoid hemorrhage, a brain contusion and edema. The patient's status deteriorated after admission, and a second CT revealed an intracerebral hematoma and marked mass effect. Transcranial color sonography revealed no flow in the ipsilateral middle and anterior cerebral arteries, which was consistent with ischemic stroke. This case demonstrates that transcranial color Doppler sonography performed early after head injury may be useful in detecting associated occlusion of cerebral vessels. PMID- 11483417 TI - Coronary artery stenoses: a phantom study using contrast enhanced three dimensional electron beam tomography. AB - This paper evaluated the accuracy of electron beam tomographic angiography (EBA) with conventional coronary arteriography (CCA) using four graded artificial stenoses in a postmortem swine coronary phantom model. The sensitivity, specificity, and accuracy of EBA for diagnosing significant stenosis (> or =50% stenosis) were 94.3%, 96.7%, and 95.8%, respectively. The diagnostic accuracy of EBA had no significant difference with CCA (chi(2)=0.0162; P>.05). EBA three dimensional (3D) procedures had high interobserver reproducibility (k=.92-.95, P>.05). Maximum intensity projection (MIP) was the most sensitive and curved planar reformation (CPR) was the most accurate 3D procedure for quantitatively identifying coronary stenosis. EBA yields promising results concerning the visualization of coronary artery stenosis with high accuracy for stenoses >50%. PMID- 11483418 TI - Duodenal hemangiopericytoma: CT and MRI findings. AB - Hemangiopericytoma (HPC) is a rare mesenchymal tumor generally occurring in adults and originating from the pericytes. The tumor more commonly affects the soft tissues of the extremities, the pelvis, and the retroperitoneum. We describe the computed tomographic and magnetic resonance appearance of a patient affected by HPC of the duodenum. PMID- 11483419 TI - Comparison of various techniques of iron oxide-enhanced breath-hold MR imaging of hepatocellular carcinoma. AB - The purpose of our study is to compare qualitatively and quantitatively the abilities of various superparamagnetic iron oxide (SPIO)-enhanced breath-hold magnetic resonance imaging (MRI) techniques to detect hepatocellular carcinoma (HCC). Eight patients with HCCs were imaged. The images were obtained with conventional T2-weighted spin-echo imaging (CSE), half-Fourier single-shot turbo spin-echo (HASTE), single-shot gradient-echo type echo planar imaging (GE-EPI), and single-shot spin-echo type echo planar imaging (SE-EPI) before and after SPIO administration. The liver signal-to-noise ratios (SNRs) in CSE and each EPI sequence were significantly decreased after SPIO administration. GE-EPI had the highest decrease ratio (DR) of liver SNR, followed by SE-EPI (TE=98), SE-EPI (TE=28), CSE, and HASTE in this order. The relative contrasts with GE-EPI and SE EPI (TE=98) were significantly higher than that with CSE after SPIO administration. On receiver operating characteristic (ROC) analysis, diagnostic accuracy did not differ significantly among the pulse sequences after SPIO administration. GE-EPI and SE-EPI (longer TE) were useful for SPIO-enhanced breath-hold MRI performed to detect HCC. PMID- 11483420 TI - MR diffusion-weighted imaging of kidney: differentiation between hydronephrosis and pyonephrosis. AB - The objective of the study was to evaluate the capability and reliability of the magnetic resonance (MR) diffusion-weighted imaging (DWI) in differentiation between hydronephrosis and pyonephrosis. Single-shot echoplanar MR diffusion weighted imaging was performed in 12 patients who had dilatation of the renal pelvis and calyces detected by ultrasonography (US). Microbiological tests confirmed that there were four cases of pyonephrosis and eight cases of hydronephrosis. Signal intensities of the collecting (pelvicalyceal) systems on the diffusion-weighted images and apparent diffusion coefficient (ADC) maps were noted. ADC values of the pelvicalyceal system in all patients were computed and compared using Student's t test. On diffusion-weighted images, the pelvicalyceal system of the hydronephrotic kidney was hypointense while the pelvicalyceal system of the pyonephrotic kidney was markedly hyperintense. The mean ADCs of the hydronephrotic and pyonephrotic renal pelvis were 2.98 +/- 0.65 x 10(-3) and 0.64 +/- 0.35 x 10(-3) mm(2)/s, respectively. The extremely low ADC of the renal pelvis of the pyonephrotic kidney accounted for its signal hyperintensity on diffusion-weighted images as well as signal hypointensity on ADC maps. In conclusion, the MR diffusion-weighted imaging may be a reliable tool to differentiate pyonephrosis from hydronephrosis. PMID- 11483421 TI - MRI isolation of infected renal cyst in autosomal dominant polycystic kidney disease. AB - The use of gadolinium-enhanced magnetic resonance (MR) to isolate an infected renal cyst in the setting of autosomal dominant polycystic kidney disease (ADPKD) has not been previously described. A case in which T1- and T2-weighted and gadolinium-enhanced MR images were used to identify a single purulent cyst in a patient with ADPKD is herein presented. We suggest that gadolinium-enhanced MR be considered useful in the evaluation of ADPKD patients with suspected infected cyst. PMID- 11483422 TI - US and CT findings of xanthogranulomatous pyelonephritis. AB - Ultrasonographic and computed tomographic findings of 21 cases of pathologically proven xanthogranulomatous pyelonephritis (XGP) in 20 patients were retrospectively evaluated. Seventeen (81%) were diffuse, and extrarenal extension occurred in 13 cases (62%). The kidney was enlarged diffusely in 12 cases (57%), and focally in 3 (14%). Urinary calculi were present in 16 cases (76%), with staghorn calculi in 4 of these, and hydronephrosis occurred in 17 (81%). In addition to typical features of XGP, the condition may also show variable imaging findings. PMID- 11483423 TI - Central lucency of pelvic phleboliths: comparison of radiographs and noncontrast helical CT. AB - To compare how often the central lucency of pelvic phleboliths is seen on plain radiographs and noncontrast helical computed tomography (CT), both images in 70 patients with renal colic were analyzed. Both images revealed the same number of phleboliths in all cases except one in which one of two phleboliths demonstrated by CT was not seen on radiographs. Radiography revealed central lucency in 95 of 149 pelvic phleboliths (63%), but CT failed to depict a hypodense center in any of 150 phleboliths. The presence or absence of central lucency on CT images cannot, therefore, be used to differentiate phleboliths from distal ureteral calculi. PMID- 11483424 TI - A true aneurysm of the femoral artery in acromegaly. AB - We report on an acromegalic patient who developed a true aneurysm in the right femoral artery. Plain computed tomography (CT) showed a soft tissue mass with low density adjacent to the right femoral artery. T1-weighted magnetic resonance (MR) images exhibited the lesion with moderate hypointensity peripherally, and hypointensity centrally. T2-weighted images showed the lesion to have areas of hypointensity, moderate hyperintensity, and marked hyperintensity. Surgery revealed an ectasia of the common femoral artery and a true aneurysm from the lateral femoral circumflex artery. PMID- 11483425 TI - Arteriovenous malformation in a patient with Bannayan--Zonana syndrome. AB - Bannayan-Zonana syndrome (BZS) is a genetic disorder with autosomal dominant inheritance characterized by macrocephaly and multiple hamartomas of mesodermal origin. Here we present a patient with BZS manifested by many of the classic features, as well as a high-flow upper extremity arteriovenous malformation (AVM). Although this rare syndrome was initially described in 1971, to our knowledge, this is the first report showing an association of AVM with BZS and the first report of this syndrome in the radiologic literature. PMID- 11483426 TI - Failure of Wallstents in the subclavian vein due to stent damage. AB - Subclavian vein stenosis and thrombosis are common problems in hemodialysis patients and in the Paget--von Schrotter syndrome. Besides surgery, several less invasive strategies as balloon angioplasty, drug-induced or mechanical thrombolysis and stenting are used in the treatment of this condition. Three cases of recurrent venous obstruction of the subclavian vein treated with placement Wallstents are described. In all three patients, rethrombosis occurred due to stent failure, two involving stent fragmentation and one, stent collapse. PMID- 11483427 TI - CT and MRI findings in sclerodermatous chronic graft vs. host disease. AB - Sclerodermatous chronic graft vs. host disease is a well-documented complication of bone marrow transplantation, however has not previously been described in the radiology literature. We present the imaging findings of this disease including the ability to identify active disease and aid in clinical management. PMID- 11483428 TI - Multiple myeloma: short T2 on MR imaging. AB - We describe a rare case of multiple myeloma with marked hypointense areas on both T1- and T2-weighted images. Amyloid deposition and calcification were not found histologically. Hypointensity on T2-weighted images may be caused by distributed free radicals produced by significant macrophages. PMID- 11483430 TI - Partial characterization of oligosaccharides expressed on midgut microvillar glycoproteins of the mosquito, Anopheles stephensi Liston. AB - Midguts of the malaria-transmitting mosquito, Anopheles stephensi, were homogenized and microvillar membranes prepared by calcium precipitation and differential centrifugation. Oligosaccharides present on the microvillar glycoproteins were identified by lectin blotting before and after in vitro and in situ treatments with endo- and exo-glycosidases. Twenty-eight glycoproteins expressed a structurally restricted range of terminal sugars and oligosaccharide linkages. Twenty-three glycoproteins expressed oligomannose and/or hybrid N linked oligosaccharides, some with alpha1-6 linked fucose as a core residue. Complex-type N-linked oligosaccharides on eight glycoproteins all possessed terminal N-acetylglucosamine, and alpha- and beta-linked N-acetylgalactosamine. Eight glycoproteins expressed O-linked oligosaccharides all containing N acetylgalactosamine with or without further substitutions of fucose and/or galactose. Galactosebeta1-3/4/6N-acetylglucosamine-, sialic acidalpha2 3/6galactose-, fucosealpha1-2galactose- and galactosealpha1-3galactose- were not detected. Terminal alpha-linked N-acetylgalactosamine residues on N-linked oligosaccharides are described for the first time in insects. The nature and function of these midgut glycoproteins have yet to be identified, but the oligosaccharide side chains are candidate receptors for ookinete binding and candidate targets for transmission blocking strategies. PMID- 11483431 TI - Structural and functional conservation and divergence among acyl-CoA desaturases of two noctuid species, the corn earworm, Helicoverpa zea, and the cabbage looper, Trichoplusia ni. AB - In this report, we describe the structural and functional analyses of four acyl CoA desaturase-encoding cDNAs that we isolated from RNA expressed in the pheromone gland of the corn earworm, Helicoverpa zea. We deduced the homology relationships of the encoded proteins, designated HzPGDs1, HzPGDs2, HzPGDs3 and HzFBDs, to each other and to previously described desaturases of the cabbage looper moth, Trichoplusia ni, the fly, Drosophila melanogaster, and other more distantly related organisms. We also isolated genomic DNA fragments of the four H. zea desaturase-encoding genes, determined the locations of introns present in them, and compared them to conserved intron positions in reported desaturase genes of other species. We measured the levels of the four desaturase mRNAs in H. zea pheromone glands and larval fat bodies by RT-PCR. We established the functional identities of the deduced proteins HzPGDs1 and HzPGDs2, encoded by the two desaturase mRNAs that are differentially and abundantly expressed in pheromone glands of sexually mature adult H. zea females, by functional expression of their encoding cDNAs in a desaturase-deficient mutant, ole1, of the yeast Saccharomyces cerevisiae. We compared the unique unsaturated fatty acid profiles of HzPGDs1- and HzPGDs2-expressing transformants to those of strains expressing previously described Delta11 and Delta9 desaturases of T. ni. PMID- 11483432 TI - Tentative identification of a resilin gene in Drosophila melanogaster. AB - A search of the Drosophila genome for gene products with similarities to the amino acid sequences of three tryptic peptides from locust (Schistocerca gregaria) resilin gave two positive results: gene products CG15920 and CG9036. In both conceptual translation products a 62-residue region is present, which is identical to the resilin peptides in 29 positions. Gene product CG15920 has an amino acid composition closely resembling that of resilins from various insect species, and it has an N-terminal signal peptide sequence indicating that it is an extracellular protein. The 62-residue region shows similarity to the RR-2 sequence, which is common for a number of matrix proteins from insect solid cuticle. The N- and C-terminal regions flanking the 62-residue in CG15920 are dominated by 18 repeats of a 15-residue sequence and 11 repeats of a 13-residue sequence, respectively. The structures of the repeats predict that the peptide chain will fold in an irregular, extended beta-spiral, resembling the structures suggested for mammalian elastin and spider flagelliform silk, two materials which, like resilin, possess long-range elasticity. Accordingly, we suggest that gene product CG15920 is a Drosophila resilin precursor. PMID- 11483433 TI - Identification of a receptor type guanylyl cyclase in the antennal lobe and antennal sensory neurons of the silkmoth, Bombyx mori. AB - Sensory neuron-specific guanylyl cyclases (GC), which were recently identified in rodents and nematodes, are thought to be a new family of odorant/pheromone receptors. In the antennae of the male silkmoth Bombyx mori, receptor type GC are supposed to mediate signaling of pheromone. Structure of receptor type GC expressed in insect sensory neurons has remained unidentified. Here we report the isolation of cDNA of the receptor type GC, designated BmGC-I, from the male silkmoth antennae. The deduced amino acid sequence indicates that BmGC-I appears to consist of four domains: an extracellular, single transmembrane, kinase-like and a guanylyl cyclase domain. BmGC-I is most closely related to the mammalian natriuretic peptide hormone receptor A (GC-A) and retains all the cysteine residues that are conserved within the extracellular domain of the mammalian GC As. Transcripts of the BmGC-I gene were detected in various tissues; the flight muscles, midgut, legs, ganglion, Malpighian tubules, testis and the head. Immunohistochemical study revealed that the BmGC-I protein localizes in the antennal-lobe glomerulus and in the soma and axon of sensory neurons. We thus suggest that BmGC-I plays functional roles in the odorant information processing and the modulation of excitability in the antennal sensory neurons. PMID- 11483434 TI - The receptor of Bacillus sphaericus binary toxin in Culex pipiens (Diptera: Culicidae) midgut: molecular cloning and expression. AB - Culex pipiens larval midgut is the primary target of the binary toxin (Bin) present in parasporal inclusions of Bacillus sphaericus. Cpm1, a 60-kDa protein purified from brush border membranes, has been proposed as the receptor of the Bin toxin in the midgut epithelial cells of mosquitoes. We have cloned and characterized the corresponding cDNA from midgut of Culex pipiens larvae. The open reading frame predicted a 580 amino-acid protein with a putative signal peptide at the N-terminus and a putative GPI-anchoring signal at the C-terminus. The amino acid sequence of the cloned Cpm1 exhibited 39-43% identities with insect maltases (alpha-glucosidases and alpha-amylases). Recombinant Cpm1 expressed in E. coli specifically bound to the Bin toxin and had a significant alpha-glucosidase activity but no alpha-amylase activity. These results support the view that Cpm1 is an alpha-glucosidase expressed in Culex midgut where it constitutes the receptor for the Bin toxin. To date, this is the first component involved in the mosquitocidal activity of the Bacillus sphaericus Bin toxin to be characterized. Its identification provides a key step to elucidate the mode of action of the Bin toxin and the mechanisms of resistance developed against it by some mosquito strains. PMID- 11483435 TI - Fenitroxon insensitive acetylcholinesterases of the housefly, Musca domestica associated with point mutations. AB - The cDNA of AChE in the housefly, Musca domestica, was sequenced and individual flies were genotyped by this gene in an inhibition assay of AChE activity with an organophaspate, fenitroxon. Mutations at Gly(342) and Tyr(407), which are reportedly conserved in resistant strains of Drosophila, were associated with the insensitivity to fenitroxon. Two other mutations, Ile(162) and Val(260), did not have an apparent effect on insensitivity. However, the four mutations are located in the active site of the enzyme, and therefore the non-neutral mutations in this gene are considered to cause the insensitivity of AChE in the development of insecticide resistance of the housefly. PMID- 11483437 TI - Purification and characterization of fibroin from the tropical Saturniid silkworm, Antheraea mylitta. AB - The fibroin protein isolated from the posterior silkgland of the tropical Saturniid silkworm Antheraea mylitta, was solubilized in lithium dodecyl sulfate and purified by gel filtration. The major fraction from gel filtration was analyzed by SDS-PAGE under non-reducing and reducing conditions. One major protein band of ca 395 kDa was obtained under non-reducing conditions and a doublet band of approximately 197 kDa under reducing conditions. The appearance of a single spot in two-dimensional electrophoresis confirmed the purity of the protein indicating that it may be a homodimeric protein of two similar sized polypeptides. Amino acid composition analysis showed that, like other Saturniid fibroins, it is rich in glycine, alanine and serine amino acids. N-terminal amino acid sequence shows significant homology with other Antheraea species. The enzymatic deglycosylation analysis indicates that the fibroin protein is glycosylated and the oligosaccharides are O-linked to the protein backbone by N acetylgalactoseamine moiety which conforms to a Core 1 mucin-type glycosylation pattern. PMID- 11483436 TI - Molecular analysis of multiple CYP6B genes from polyphagous Papilio species. AB - Papilio glaucus (eastern tiger swallowtail) and Papilio. canadensis (Canadian tiger swallowtail) are two closely related species with broad but overlapping hostplant ranges. P. glaucus encounters toxic furanocoumarins occasionally in its diet in its rutaceous hostplants, whereas P. canadensis rarely if ever encounters these compounds. Analysis of their furanocoumarin-metabolic profiles indicates that these species induce cytochrome P450 monooxygenases (P450s) capable of metabolizing linear and angular furanocoumarins to varying degrees in response to dietary supplementation with xanthotoxin (a linear furanocoumarin). In P. glaucus, metabolism is induced to a significantly higher level than in P. canadensis. Cloning of multiple P450 genes from each species has revealed that both species contain and express two groups of P450s, designated CYP6B4 and CYP6B17, that are related to the P. glaucus CYP6B4v1 enzyme known to metabolize an array of furanocoumarins. Expression patterns of the CYP6B4 and CYP6B17 group transcripts differ in these species in both their basal and furanocoumarin inducible levels. In P. glaucus, CYP6B4 transcripts, which are not detectable constitutively, are 311-fold induced by xanthotoxin and CYP6B17 transcripts, which are detectable constitutively, are 3-fold induced by xanthotoxin. In P. canadensis, CYP6B4 transcripts are only 8-fold induced and CYP6B17 transcripts are 13-fold induced. These findings are consistent with the postulated evolutionary history of these two species, according to which P. glaucus maintains its association with rutaceous hostplants and P. canadensis has differentiated to utilize hostplants in other families more extensively. PMID- 11483438 TI - Mass isolation of cuticle protein cDNAs from wing discs of Bombyx mori and their characterizations. AB - Multiple cloning of cuticle protein genes was performed by sequencing of cDNAs randomly selected from a cDNA library of wing discs just before pupation, and nine different cuticular protein genes were identified. Thirty-one clones of a cuticle protein gene were identified from the 1050 randomly sequenced clones; about 3% were cuticle protein genes in the W3-stage wing disc cDNA library. The sequence diversity of the deduced amino acid sequences of isolated Bombyx cuticle genes was examined along with the expression profiles. The deduced amino acid sequences of the nine cuticle protein genes contained a putative signal peptide at the N-terminal region and a very conserved hydrophilic region known as the R and R motif. The developmental expression of cuticle genes was classified into two types: pupation (five clones were expressed only around pupation) and pupation and mid-pupal (four clones were expressed around this stage). All the isolated genes were expressed in the head, thoracic, and abdominal regions of the epidermis at different levels around pupation, but no expression was observed in the epidermis at the fourth molting stage. PMID- 11483439 TI - The esophagus. AB - The intent of this article is to raise the clinician's awareness of manifestations of common esophageal abnormalities and disorders of swallowing. The specifics of esophageal disorders are reviewed. PMID- 11483440 TI - Peptic ulcer disease. AB - Peptic ulcer disease is a common gastrointestinal disease whose management and treatment has changed dramatically over the last 25 years. Treatment of peptic ulcer disease has evolved from dietary modifications and antacids to gastric acid suppression with H2-receptor antagonists and proton pump inhibitors to eradication of Helicobactor pylori infection. Treatment of patients infected with H pylori using antibiotics has changed the natural history of peptic ulcer disease. As a result of H pylori treatment and other unknown factors ulcer disease is declining and complications from ulcer disease have diminished significantly. PMID- 11483441 TI - Malabsorption and malnutrition. AB - In this article, disease of the small intestine will be discussed, with particular reference to those conditions leading to malabsorption and malnutrition. The work up of these entities will be emphasized, with focus on bacterial overgrowth, celiac sprue and nonsteroidal-induced enteropathy. PMID- 11483442 TI - Irritable bowel syndrome. AB - Irritable bowel syndrome is a common gastrointestinal disorder characterized by abdominal pain and a change in bowel habits. It is estimated to occur in 9% to 22% of the US population, and is more often diagnosed in women than in men. There are no diagnostic markers for irritable bowel syndrome and no evidence of organic disease. Moreover, the pathophysiology is not completely understood; however, abnormal gastrointestinal motility, altered sensory activity, central neural dysfunction, psychological disturbances, and luminal factors have been proposed. Treatment requires a multifaceted approach. Education, dietary measures, fiber supplementation, and pharmacological and psychological interventions have been used in the management of this disorder. PMID- 11483443 TI - Inflammatory bowel disease. AB - Idiopathic inflammatory bowel disease consists of Crohn's disease and ulcerative colitis. Crohn's disease can affect any part of the gastrointestinal tract, from the mouth to the anus, and is also known as regional enteritis, terminal ileitis, or granulomatous colitis. Ulcerative colitis is limited to the colon and rectal involvement is present 95% of the time. Ten percent to fifteen percent of patients with irritable bowel syndrome cannot be clearly defined as having either Crohn's disease or ulcerative colitis and are termed indeterminate colitis. PMID- 11483444 TI - Management principles of gastrointestinal bleeding. AB - There is now a multitude of technologic and pharmacologic options available to clinicians caring for patients with gastrointestinal (GI) bleeding; however, drugs and technology are no substitute for understanding and properly executing the basic management principles of GI bleeding. This article focuses on the most common causes of GI bleeding and emphasizes the importance of the primary care provider's role in the management of these patients. Also, by answering questions we are commonly asked as gastroenterology consultants, we hope to provide insight into current diagnostic and therapeutic options and the most appropriate use of these options. PMID- 11483445 TI - Constipation: evaluation and management. AB - Constipation is a common symptom that can result from several disorders. Most patients with chronic constipation do not have a medical disorder contributing to the constipation and, therefore, require limited evaluation. Initial intervention should include dietary measures and fiber supplements; however, if fiber supplementation is ineffective, other agents can be used. Surgery should be reserved for patients who meet specific clinical criteria. PMID- 11483446 TI - Gallstones and biliary disease. AB - Gallstones are common in the US and western countries. This article describes the pathogenesis of gallstone formation and the clinical manifestations and current approaches to diagnosis and treatment of the most common clinical conditions caused by gallstones: biliary colic, acute cholecystitis, choledocholithiasis, and acute gallstone pancreatitis. The role of widely used imaging techniques (transabdominal ultrasound, CT scan, MR imaging, and MRCP) and diagnostic and therapeutic endoscopy (endoscopic ultrasound, ERCP) is emphasized. This article is intended mainly for general practitioners, primary care physicians, and other specialists providing medical care to patients with gallstones and their complications. PMID- 11483447 TI - Acute and chronic pancreatitis. AB - Acute pancreatitis has multiple causes, an unpredictable course, and myriad complications. The diagnosis relies on a combination of history, physical examination, serologic markers, and radiologic findings. The mainstay of therapy includes aggressive hydration, maintenance of NPO, and adequate analgesia with narcotics. Antibiotic and nutritional support with total parenteral nutrition should be used when appropriate. PMID- 11483448 TI - Nutrition in gastroenterology and hepatology. AB - Nutritional issues, sometimes obvious and sometimes not so obvious, confront the primary care practitioner on a daily basis. Understanding the multi-disciplinary nature of nutrition science and clinical nutrition and having a basic understanding of gastrointestinal tract digestive and absorptive physiology can only help sort out the many issues involved. The general practitioner needs a core understanding of the principles of nutrition assessment. PMID- 11483449 TI - Gastrointestinal malignancies. AB - Gastrointestinal cancers are collectively the most common malignancies in the world. In the United States, colorectal cancer accounts for most of these malignancies. This is unfortunate, because colorectal cancer is highly preventable. Moreover, screening for colorectal cancer is cost effective and becomes increasingly important in an aging population. Individuals who are more at risk require earlier screening and intervention. For all gastrointestinal cancer, surgery remains the most important curative option. For patients who cannot be cured, quality of life is the yardstick by which palliative options should be evaluated. PMID- 11483450 TI - Colorectal cancer screening. AB - Colorectal cancer causes significant morbidity and mortality in the United States. The incidence of colorectal cancer increases at age 50, approximately. Risk factors that have been identified include a personal history of colorectal cancer or adenomas, a family history of colon cancer or adenomas, inherited colorectal cancer syndromes, and long standing inflammatory bowel disease. Several screening tests have been developed for colorectal cancer prevention. Surveillance strategy is based on an individual's colorectal cancer risk. This article reviews fecal occult blood testing, flexible sigmoidoscopy, colonoscopy, barium enema, and genetic testing. PMID- 11483451 TI - Effects of biphasic vs monophasic defibrillation on the scaling exponent in a swine model of prolonged ventricular fibrillation. AB - OBJECTIVE: Mathematical analyses of ventricular fibrillation (VF) have resulted in the derivation of a measure termed the scaling exponent (ScE) that characterizes the duration of VF and probability of defibrillation success. The purpose of this study was to compare the effects of biphasic defibrillation waveform (BDW) and monophasic defibrillation waveform (MDW) rescue shocks on ScE in a swine model of prolonged VF. METHODS: Utstein guidelines for the laboratory study of cardiopulmonary resuscitation were followed. Twenty mixed-breed domestic swine (mass range 20.5-26.8 kg) were instrumented and randomized to receive either MDW or BDW rescue shocks. Ventricular fibrillation was induced and untreated for a nonintervention interval of 8 minutes. Rescue shocks were delivered at 8, 10, and 12 minutes of elapsed VF time. The energy sequence for the three MDW shocks was 70, 100, and 150 J (approximately 3, 4, and 6 J/kg). All BDW shocks were delivered at 50 J (approximately 2.5 J/kg). Only VF was shocked. Chest compressions and drugs were not provided. Rhythm analysis and ScE calculation were performed offline. Continuous and discontinuous linear regression models were fit to plots of ScE vs time. Defibrillation success and progression of ScE slope were analyzed using Fisher's exact test, paired t-tests, and repeated-measures analysis of variance (ANOVA). RESULTS: Baseline characteristics were similar for both groups. Successful termination of VF occurred on the first rescue shock in 1 of 10 (10%) in the MDW group and 3 of 10 (30%) in the BDW group; this difference was not statistically significant (p = 0.58). No other defibrillation successes were observed. No animals achieved return of spontaneous circulation. The ScE values during the protocol progressed from 1.330 (95% CI = 1.287 to 1.373) to 1.724 (95% CI = 1.603 to 1.845) for MDW and 1.338 (95% CI = 1.261 to 1.415) to 1.639 (95% CI = 1.530 to 1.745) for BDW. Both groups showed a trend toward increasing ScE values with successive rescue shocks. Repeated-measures ANOVA using both continuous and discontinuous models demonstrated no difference in overall ScE slope progression between study groups. CONCLUSIONS: Mode of defibrillation waveform (BDW vs MDW) does not appear to impact ScE trends. Additional studies must be performed to better evaluate the clinical implications of this finding. PMID- 11483452 TI - Children with first-time simple febrile seizures are at low risk of serious bacterial illness. AB - OBJECTIVE: To describe the rates of serious bacterial illness (SBI) in children presenting to emergency departments (EDs) with first-time uncomplicated febrile seizures. METHODS: The ED visits from seven Chicago metropolitan area hospitals (two tertiary pediatric EDs, five community general EDs) for all pediatric patients seen between July 1995 and December 1997 with a discharge diagnosis including the term "seizure" were retrospectively identified. Records of patients who met criteria for simple, first-time febrile seizure were reviewed (age 6-60 months; temperature > or =38.0 degrees C; single, generalized, tonic-clonic seizure <20 minutes; absence of known central nervous system disease). Rates of bacteremia, urinary tract infection, bacterial meningitis, and pneumonia were determined. RESULTS: Four hundred fifty-five children were identified who had first-time simple febrile seizures. The study participants had a mean age of 21 months and a mean temperature of 39.6 degrees C, and 64% were male. Seventy-three percent were seen in a community hospital setting. Blood cultures were obtained for 315 children (69%). Four children (1.3% [95% CI = 0.1% to 2.5%]) were bacteremic, all with Streptococcus pneumoniae; the rate of bacteremia did not differ in the subset at highest risk for bacteremia (6-36 months, temperature >39 degrees C). No demographic or laboratory data distinguished the bacteremic children from those with negative blood cultures. One hundred seventy-one children (38%) had urine cultures obtained; 5.9% [95% CI = 2.4% to 9.4%] of the cultures grew >100,000 colony-forming units/mL of a single pathogenic organism. One hundred thirty-five children (30%) had cerebrospinal fluid cultures performed. None of these cultures grew a bacterial pathogen [95% CI = 0% to 2.2%]. Two hundred eight children (45.7%) had chest x-rays performed; 12.5% [95% CI = 10.2% to 14.8%] (n = 26) of the x-rays were read as consistent with pneumonia by the radiologist at the treating institution. None of the blood cultures performed on children with abnormal radiographs were positive (cultures drawn on 23 of 26 patients, 88%). Stool cultures were performed on 14 children (3.1%); two cultures (14.3% [95% CI = 0% to 32.6%]) grew a bacterial pathogen, both Shigella. CONCLUSIONS: Rates of SBI in this multi-institution population of children with first-time simple febrile seizures were low and are consistent with those published in the literature for febrile children without seizures. PMID- 11483453 TI - Predicting postconcussion syndrome after minor traumatic brain injury. AB - BACKGROUND: Up to 50% of patients with minor traumatic brain injury (mTBI) develop postconcussion syndrome (PCS). A decision rule to stratify risk for PCS is needed. OBJECTIVE: To identify mTBI patients at low and high risk of PCS by comparing the predictive values of variables generated by logistic regression (LR) and recursive partitioning (RP). METHODS: This was a prospective, observational study of 69 mTBI patients aged >16 years presenting to the emergency department of a university teaching hospital. Minor TBI was defined as loss of consciousness <10 minutes or amnesia, Glasgow Coma Scale score (GCS) of 15, no skull fracture on physical examination, nonfocal neurologic exam, and no brain injury on computed tomography if one was done. Clinical/demographic data and the results of a brief neurobehavioral test battery were collected for all patients. The presence of PCS was determined by a validated telephone questionnaire at one month after initial presentation. All variables were subjected to both LR and RP. RESULTS: Fifty-eight percent had PCS at one month after initial presentation. Low risk: PCS occurred in 9% of men scoring >24 on the Hopkins Verbal Learning A (HVLA) (by LR) and in 9% of those injured in sports scoring >22 on HVLA (RP). High risk: PCS occurred in 89% of women scoring <9 on the Digit Span test (LR) and in 92% of those injured via falls or motor vehicle collision scoring <11.5 on HVLB2 (RP). CONCLUSIONS: Despite the high incidence of PCS, we were able to identify a low-risk subgroup with an average PCS risk of <10% and a high-risk subgroup with a PCS risk of approximately 90%. Combining results from LR and RP expanded the number of patients able to be classified as high/low risk. Prospective validation is necessary. PMID- 11483454 TI - Domestic abuse in the emergency department: can a risk profile be defined? AB - OBJECTIVES: The main purpose of this study was to determine whether any clinical or demographic characteristics could identify adult female patients presenting to the emergency department (ED) with a history of domestic abuse. A second objective was to describe the frequency, types, and severity of this abuse. METHODS: This study was a crosssectional survey of 611 women conducted in an academically-affiliated, urban ED. Domestic abuse was described as "recent" (within the preceding 12 months) or "lifetime" (recent or past). This included emotional, physical, and sexual abuse. RESULTS: Recent (7.9%, n = 48) and lifetime (38%, n = 232) domestic abuse was reported. For recently abused women, violence had been severe (87.5%, n = 42) and was associated with 1) trauma (OR 5.4, 95% CI = 2.6 to 11.6), 2) obstetrical and gynecological syndromes (OR 5.6, 95% CI = 2.4 to 13.2), and 3) psychiatric symptoms and substance use (OR 7.3, 95% CI = 2.4 to 22.0). The sensitivities and positive predictive values of these risk factors individually (<27.1% and <25.0%, respectively) and in aggregate (56.3% and 20.9%, respectively) were low. These indicators predicted only 27 (56.3%) of recently abused women. Lifetime domestic violence was more likely in homeless women (OR 5.8, 95% CI = 2.2 to 15.0), although less likely in immigrants (OR 0.4, 95% CI = 0.3 to 0.7). CONCLUSIONS: Clinical presentations and demographic characteristics of women presenting to the ED may not be sensitive or predictive indicators of domestic abuse. In the absence of typical clinical or demographic findings, asking all women in the ED about domestic abuse remains a necessary priority. PMID- 11483455 TI - Impact of a resident strike on emergency department quality indicators at an urban teaching hospital. AB - OBJECTIVE: To evaluate the indicators of activity and quality within the emergency department (ED) during a resident physicians' strike. METHODS: This was an observational study comparing a strike period (SP) and a non-strike period (NSP) in the ED of a 1,000-bed tertiary care teaching hospital in Barcelona, Spain, with an annual census of 100,000 emergency visits. During a period of nine nonconsecutive days, the resident physicians were on strike. Emergency visits were handled by staff members. Data were compared between all patients treated in the ED during the SP and those treated during the NSP, matched by the weekday. The authors compared lengths of stay (LOSs), rates of use of laboratory tests and radiology procedures, numbers of patient walkouts, patient/physician ratios, emergency hospital admission rates, home discharge rates, unscheduled return rates, and mortality rates. RESULTS: The two groups (SP 2,610 patients and NSP 3,634 patients) were comparable in terms of average daily attendance rate (SP: 290 +/- 12 vs NSP: 302 +/- 21; p = 0.13), elective hospital admission rate, and severity of illness. Statistically significant differences were found in terms of mean total patients' LOS (SP: 206.75 +/- 12.27 vs NSP: 235.10 +/- 27.08 minutes; p < 0.001), number of laboratory tests per patient (SP: 0.30 +/- 0.05 vs NSP: 0.38 +/- 0.04; p < 0.001), and radiographs per patient (SP: 0.78 +/- 0.06 vs NSP: 0.88 +/- 0.09; p = 0.021). CONCLUSIONS: This study demonstrated that replacing residents with staff physicians resulted in fewer laboratory tests ordered, fewer radiographs ordered, and shorter lengths of stays in the ED. PMID- 11483457 TI - Report from the Council of Emergency Medicine Residency Directors subcommittee on graduate medical education funding: effects of decreased medicare support. AB - INTRODUCTION: Recent changes by the Health Care Financing Administration (HCFA) have resulted in decreased Medicare support for emergency medicine (EM) residencies. OBJECTIVE: To determine the effects of reduced graduate medical education (GME) funding support on residency size, resident rotations, and support for a fourth postgraduate year (PGY) of training and for residents with previous training. METHODS: A 36-question survey was developed by the Council of Emergency Medicine Residency Directors (CORD) committee on GME funding and sent to all 122 EM program directors (PDs). Responses were collected by the Society for Academic Emergency Medicine (SAEM) office and blinded with respect to the institution. RESULTS: Of 122 programs, 109 (89%) responded, of which 78 were PGY 1-3 programs, 19 were PGY 2-4, and 12 were PGY 1-4. The PDs were asked specifically whether there were changes in program size due to changes in Medicare reimbursement. Although few programs (12%) decreased their size or planned to decrease their size, 39% had discussions regarding decreasing their size. Thirty percent of the PDs responded that other programs at their institution had already decreased their size; 26% of the PDs had problems with financing outside rotations; and 24% had a decrease in off-service residents in their emergency departments (EDs). Only seven (6%) of programs paid residents from practice plan dollars, while most (82%) were fully supported by federal GME funding. Nearly all four-year programs (97%) received full resident salary support from their institutions and 77% of programs accept residents with previous training. CONCLUSIONS: Nearly all EM programs are fully supported by their institutions, including the fourth postgraduate year. Most programs take residents with previous training. Although few programs have reduced their size, many are discussing this. Many programs have had difficulty with funding off service rotations and many have had decreased numbers of off-service residents in their EDs. Recent GME funding changes have had adverse effects on EM residency programs. PMID- 11483458 TI - Initial experience of emergency physicians using the intubating laryngeal mask airway: a case series. AB - OBJECTIVE: Although the intubating laryngeal mask airway (ILMA) is widely available, its use by emergency physicians (EPs) has not been reported. The authors report the initial experience of EPs using the ILMA. A review of their experience and the relevant anesthesia literature provides a basis for EPs to use the ILMA more confidently and effectively. METHODS: Between January 2000 and January 2001, the ILMA was used on a convenience sample of emergency department (ED) patients undergoing "routine" intubations, and "rescue" situations, after failed rapid-sequence intubation (RSI). Patients were identified from the ED resuscitation case database. Chart review and intubating physician interviews focused on success of the device, complications encountered, and "pearls" of the device's use as perceived by the intubating physician. RESULTS: Ventilation with the appropriate-size ILMA occurred in less than 15 seconds in all "routine" intubations; tracheal intubation was subsequently accomplished in less than 1 minute. Eight of nine "routine" patients had blind tracheal intubation through the ILMA. One patient required fiberoptic bronchoscopy to guide the endotracheal tube into the trachea. Of the "rescue" intubations, all patients (n = 7) were successfully ventilated and five were successfully intubated using the ILMA. CONCLUSIONS: In this case series, the ILMA was easy to use in acute resuscitations, and proved to be invaluable in cases of failed RSI. PMID- 11483459 TI - Preventive care in the emergency department: should emergency departments institute a falls prevention program for elder patients? A systematic review. AB - OBJECTIVE: To perform a systematic review of the emergency medicine literature to assess the appropriateness of an intervention to identify, counsel, and refer emergency department (ED) patients >64 years old who are at high risk for falls. METHODS: The systematic review was facilitated through the use of a structured template, a companion explanatory piece, and a grading and methodological scoring system based on published criteria for critical appraisal. A reference librarian did two PubMed searches using the following: ED visits, patients >64 years old, falls, high risk, and effectiveness of intervention. Emergency Medical Abstracts, Science Citation Index, and the Cochrane Collaboration database were searched. Two team members reviewed the abstracts and selected pertinent articles. References were screened for additional pertinent articles. RESULTS: Twenty-six articles were reviewed. None were ED-based primary or secondary falls prevention in older patients. One randomized controlled trial of an intervention to decrease subsequent falls in elder community-dwelling patients who presented with a fall showed a structured interdisciplinary approach significantly reducing the number of falls. Two ED-based studies showed it was possible to identify ED patients at risk for falls. CONCLUSIONS: Based on one randomized controlled trial demonstrating a significant reduction in the risk of further falls, the burden of suffering caused by falls, and other studies demonstrating the value of interventions to reduce the risk of falling, the authors recommend that EDs conduct research to evaluate the effectiveness of clinical interventions to identify, counsel, and refer ED patients >64 years old who are at high risk for an unintentional fall. PMID- 11483460 TI - Clinical supervision in the emergency department: a costly inefficiency for academic medical centers. PMID- 11483461 TI - Myths and realities: the "difficult airway" and alternative airway devices in the emergency setting. PMID- 11483462 TI - The intubating laryngeal mask airway: suggestions for use in the emergency department. AB - With the increased use of rapid-sequence induction and its potential complications, emergency physicians need a rescue device for unexpected difficult intubations. The intubating laryngeal mask airway (ILMA) is an ideal rescue airway since it can be placed quickly and can provide adequate ventilation in nearly all patients. It can then be used as conduit for endotracheal intubation, while ventilation is ongoing. The authors review the current literature on the ILMA. In conjunction with their experience using the ILMA in the emergency department (ED), a modification of the American Society of Anesthesiologists difficult airway algorithm was derived for use in the ED. The ILMA appears to be valuable for managing difficult airways. PMID- 11483463 TI - Resuscitation research: future directions and ethical issues. AB - Cardiopulmonary resuscitation research is a vital area of research that has made significant contributions to medical care over the last several decades. Abundant information currently exists in the medical literature, as a result of research in the areas of cardiac arrest and outcomes, physiology of cardiac arrest, pharmacologic treatments, mechanical interventions, and societal and ethical issues. Despite numerous clinical trials demonstrating physiologic benefit of various interventions, few interventions have had as much impact on outcomes of cardiac arrest as have improvements in out-of-hospital response times and rapid availability of medical treatment. Although abundant information exists regarding physiologic aspects of resuscitation, relatively little information exists about ethical, psychological, and social aspects of resuscitation. In addition to attempts to improve outcomes of cardiac arrest, researchers should also strive to improve the experiences of patients and families involved. These realities provide future strategies and directions for the best use of resuscitation research resources; although physiologic and pharmacologic research will always have significant roles in the improvement of medical care, the rapid delivery of out-of-hospital care and ethical issues will be indispensable areas of research focus in the future. PMID- 11483464 TI - Research fundamentals: choosing an appropriate journal, manuscript preparation, and interactions with editors. AB - This article, the last in a series on research methodology prepared by the Research Committee of the Society for Academic Emergency Medicine (SAEM), describes the process of journal selection, manuscript preparation, manuscript submission, and responding to editorial and reviewer comments. Methods for completing each step of this process are described. Following these methods will increase the chance of publishing one's work in the highest-quality and most appropriate journal. PMID- 11483465 TI - Surrogate consent by family members for out-of-hospital cardiac arrest research. AB - OBJECTIVES: To evaluate the availability of family members of potential subjects to provide consent for participation in out-of-hospital cardiac arrest (OOHCA) research and to estimate the time required to contact a family member. METHODS: This study was a prospective observational study of adult patients (>18 years old) with nontraumatic OOHCA treated by an urban emergency medical service. Emergency medicine resident physicians responded to each scene and noted the presence of family members. A subsequent convenience sample of family members answered standardized questions about their ability to provide consent for research participation on behalf of the patient. RESULTS: Physicians were present at 100 of the 112 adult medical cardiac arrests during the study period. A family member was present at 57% of the scenes (95% CI = 47% to 67%). Patients with family present were older and were less likely to have bystander cardiopulmonary resuscitation (CPR) or live in a nursing home. The mean time (+/-SE) from emergency dispatch to family contact was 24.40 (+/-2.60) minutes and from physician arrival to family contact was 2.45 (+/-0.87) minutes (n = 20). Eight of 13 family members were willing to enroll the patient into a resuscitation study, but five family members were unable to understand the explanation of informed consent. CONCLUSIONS: Family members were present for an unrepresentative subset of OOHCA cases, and were contacted after the therapeutic window for many interventions. The emotional nature of the emergency situation also limited the reliability of surrogate consent for OOHCA research. PMID- 11483466 TI - An unexpected complication of central line placement. PMID- 11483467 TI - Causal processes in development and psychopathology. PMID- 11483468 TI - 'New' and 'old' antidepressants: all equal in the eyes of the lore? PMID- 11483469 TI - Specificity and heterogeneity in children's responses to profound institutional privation. AB - BACKGROUND: The sequelae of profound early privation are varied. AIMS: To delineate the behavioural patterns that are specifically associated with institutional privation. METHOD: A group of 165 children adopted from Romania before the age of 42 months were compared at 4 years and 6 years with 52 non deprived UK children adopted in infancy. Dysfunction was assessed for seven domains of functioning. The groups were compared on which, and how many, domains were impaired. RESULTS: Attachment problems, inattention/overactivity, quasi autistic features and cognitive impairment were associated with institutional privation, but emotional difficulties, poor peer relationships and conduct problems were not. Nevertheless, one-fifth of children who spent the longest time in institutions showed normal functioning. CONCLUSIONS: Attachment disorder behaviours, inattention/overactivity and quasi-autistic behaviour constitute institutional privation patterns. PMID- 11483470 TI - Child sexual abuse, poor parental care and adult depression: evidence for different mechanisms. AB - BACKGROUND: Child sexual abuse (CSA) and poor parental care (neglect and institutional care) are associated with depression in adult life. Little is known about possible mechanisms underlying these associations. AIMS: To examine the role of adult intimate-love relationships as differential mediators or moderators of the associations between CSA, poor parental care and adult depression. METHOD: Sampling was carried out in two phases. In the first, questionnaires were sent to women aged 25-36 years in five primary care practices. Second-phase subjects for interview (n=198) were drawn from three strata defined on the basis of childhood adversities. Recalled childhood experiences and recent adult relationships and depression were assessed and rated independently. Frequencies of predictor and response variables, effect estimates and their confidence intervals were weighted back to the general population questionnaire sample. RESULTS: The risk for depression associated with CSA was unaffected by quality of adult relationships, while the risk associated with poor parental care was substantially altered. CONCLUSIONS: There may be different pathways linking CSA and poor parental care to adult depression. PMID- 11483471 TI - Family settings and children's adjustment: differential adjustment within and across families. AB - BACKGROUND: Children in stepfamilies and single-parent families exhibit elevated levels of behavioural and emotional problems compared with children in intact (biological) families, but there is variation within and across these family types. AIMS: To examine the sources of variation in children's behavioural and emotional problems across diverse family settings. METHOD: Levels of behavioural and emotional problems in children from diverse stepfamilies and single-parent families were compared with children living with both biological parents. Psychosocial risks were measured at the individual child and family levels. RESULTS: Behavioural and emotional problems were elevated in children in stepmother/complex stepfamilies and single-parent families, but not in simple stepfather families, relative to 'biological' families. Psychopathology associated with family type was explained by compromised quality of the parent child relationship, parental depression and socio-economic adversity. Sibling similarity in behavioural and emotional problems was most pronounced in high-risk family settings. CONCLUSIONS: Family type is a proxy for exposure to psychosocial risks; the extent of family-wide influence on children's development may be strongest in high-stress settings. PMID- 11483472 TI - Genetic moderation of environmental risk for depression and anxiety in adolescent girls. AB - BACKGROUND: There is huge individual variation in people's response to negative life events. AIMS: To test the hypothesis that genetic factors moderate susceptibility to the environmentally mediated risks associated with negative life events. METHOD: The Virginia Twin Study of Adolescent Behavioral Development (VTSABD) was used to study the effects of independent life events (assessed from maternal interview) on depression/anxiety (assessed from child interview) in 184 same-gender female twin pairs, aged 14--7 years, measured on two occasions. RESULTS: There was no genetic effect on the independent negative life events studied. A significant gene-environment interaction was found using structural equation modelling. There was no effect of independent life events on adolescents' depression in the absence of parental emotional disorder, but a significant effect in its presence. CONCLUSIONS: There is an environmentally mediated effect of life events on depression/anxiety. Genetic factors play a significant role in individual differences in susceptibility to these environmentally mediated risks. PMID- 11483473 TI - Approaches to gene mapping in complex disorders and their application in child psychiatry and psychology. AB - BACKGROUND: Twin studies demonstrate the importance of genes and environment in the aetiology of childhood psychiatric and neurodevelopmental disorders. Advances in molecular genetics enable the identification of genes involved in complex disorders and enable the study of molecular mechanisms and gene--environment interactions. AIMS: To review the role of molecular genetics studies in childhood behavioural and developmental traits. METHOD: Molecular approaches to complex disorders are reviewed, with examples from autism, reading disability and attention-deficit hyperactivity disorder (ADHD). RESULTS: The most robust finding in ADHD is the association of a variable number tandem repeat polymorphism in exon 3 of the DRD4 gene. Other replicated associations with ADHD are outlined in the text. In autism, there is a replicated linkage finding on chromosome 7. Linkage studies in reading disability have confirmed a locus on chromosome 6 and strongly suggest one on chromosome 15. CONCLUSIONS: In the next 5--0 years susceptibility genes for these disorders will be established. Describing their relationship to biological and behavioural function will be a far greater challenge. PMID- 11483474 TI - Hyperphagic short stature and Prader--Willi syndrome: a comparison of behavioural phenotypes, genotypes and indices of stress. AB - BACKGROUND: The clinical features of hyperphagic short stature (HSS) include short stature secondary to growth hormone insufficiency, excessive appetite (hyperphagia) and mild learning disabilities. Affected children characteristically live in conditions of high psychosocial stress. Symptoms resolve when the child is removed from the stressful environment. Family studies indicate a genetic predisposition. AIMS: To compare the behavioural and stress profiles of HSS with those of Prader--Willi syndrome (PWS), and to test the hypothesis that the genetic locus that predisposes to HSS co-inherits with the PWS locus at 15q11--13. METHOD: Twenty-five children with HSS, mean age 9.1 (s.d. 3.8) years, 28% female, were compared with 30 children with PWS, mean age 8.8 (s.d. 2.8) years, 33% female. RESULTS: The clinical profiles were largely similar across the conditions, but no evidence was found in HSS of co-inheritance of the PWS critical region. CONCLUSIONS: Hyperphagic short stature is one of the very few behavioural diseases associated with a pathognomonic physiological abnormality. Investigations of the suggested genetic dysregulation, which is so sensitive to environmental influences, may well be of importance in a broader context. PMID- 11483475 TI - Neuropsychological analyses of impulsiveness in childhood hyperactivity. AB - BACKGROUND: Neuropsychological analyses of impulsiveness are needed to refine assessment of attention-deficit hyperactivity disorder (ADHD). AIMS: To investigate specific impairments in hyperactive children in a neuropsychological task battery of impulsiveness, the Maudsley Attention and Response Suppression (MARS) task battery, and to identify the neural substrates. METHOD: Impulsiveness was assessed using different tasks of inhibitory control and time management (MARS) in 55 children with ADHD, other diagnoses and controls. Functional magnetic resonance images were obtained from adolescents with and without ADHD during three of the tasks. RESULTS: Children with ADHD, but not psychiatric controls, were impaired on tests of response inhibition, but not of motor timing. Reduced right prefrontal activation was observed in hyperactive adolescents during higher level inhibition and delay management, but not during simple sensorimotor coordination. CONCLUSIONS: Attention-deficit hyperactivity disorder is characterised by specific deficits in tasks of motor response inhibition, but not motor timing, and by dysfunction of frontostriatal brain regions. PMID- 11483476 TI - Autonomic reactivity and psychopathology in middle childhood. AB - BACKGROUND: Better indicators are needed for identifying children with early signs of developmental psychopathology. AIMS: To identify measures of autonomic nervous system reactivity that discriminate children with internalising and externalising behavioural symptoms. METHOD: A cross-sectional study of 122 children aged 6--7 years examined sympathetic and parasympathetic reactivity to standardised field-laboratory stressors as predictors of parent- and teacher reported mental health symptoms. RESULTS: Measures of autonomic reactivity discriminated between children with internalising behaviour problems, externalising behaviour problems and neither. Internalisers showed high reactivity relative to low-symptom children, principally in the parasympathetic branch, while externalisers showed low reactivity, in both autonomic branches. CONCLUSIONS: School-age children with mental health symptoms showed a pattern of autonomic dimorphism in their reactivity to standardised challenges. This observation may be of use in early identification of children with presyndromal psychopathology. PMID- 11483477 TI - Timing of initial exposure to maternal major depression and children's mental health symptoms in kindergarten. AB - BACKGROUND: Little is known about the influence on child mental health symptoms of the timing of initial exposure to maternal major depression or whether the timing is associated with 'pure' or co-occurring internalising and externalising symptoms. AIMS: To address these issues, while also taking account of child gender and family socio-economic status. METHOD: In a prospective community-based study, 421 kindergarten teachers rated children's symptoms. Previous assessments of maternal major depression indicated whether children were first exposed during infancy, in the toddler/pre-school period, or never. RESULTS: Exposure during infancy was associated with high internalising symptoms, especially when co occurring with high externalising symptoms. Initial exposure in the toddler/pre school years increased the risk of 'pure' externalising symptoms among girls. CONCLUSIONS: The association of child mental health symptoms with the timing of initial exposure to maternal depression highlights the need for effective prevention and intervention strategies addressed to the developmental issues of each period. PMID- 11483478 TI - Influence of psychiatric disorder on the controlling behaviour of mothers with 1 year-old infants. A study of women with maternal eating disorder, postnatal depression and a healthy comparison group. AB - BACKGROUND: Certain styles of parental controlling behaviour influence child development. Work with mothers with eating disorders suggests that they may be particularly controlling of their infants. AIMS: To examine the nature and specificity of maternal controlling behaviour in mothers with eating disorders compared with mothers who had experienced postnatal depression and a healthy comparison group. METHOD: Mothers with eating disorders (n=34), postnatal depression (n=39) and a healthy comparison group (n=61) and their 12-month-old infants were observed during play and mealtimes, and blind ratings made of verbal and non-verbal control exerted by the mother. RESULTS: Mothers in the eating disorder group used more verbal control, especially strong control. There were no differences between the groups on gentle verbal control and physical contact. Maternal dietary restraint was the one feature of eating disorder psychopathology associated with the use of verbal control. Marital criticism was also associated with the extent of verbal controlling behaviour. CONCLUSIONS: Aspects of maternal control of infants were found to be specific to maternal eating disorder psychopathology. PMID- 11483479 TI - Use of sertraline, paroxetine and fluvoxamine by nursing women. AB - BACKGROUND: The pharmacological treatment of depression in nursing women requires information on the magnitude of medication exposure to the infant that may occur through breast milk. AIMS: To examine serum concentrations of antidepressants in infants exposed to these medications through breast-feeding. METHOD: Maternal and infant serum concentrations of sertraline, paroxetine and fluvoxamine were determined with high-performance liquid chromatography (limit of detection=1 ng/ml). RESULTS: No detectable medication was present in any infant exposed to paroxetine (n=16) or fluvoxamine (n=4). Among infants exposed to sertraline (n=30), detectable medication was present in 24% of serum samples. A significant negative correlation was found between infant age and infant serum concentration. Sertraline was significantly more likely to be detected in an infant if the mother's daily dose was 100 mg or higher. No adverse sequelae occurred in any infant. CONCLUSIONS: This study shows that paroxetine, fluvoxamine and sertraline produce minimal exposure to infants when taken by nursing mothers. PMID- 11483480 TI - Association of autonomic dysfunction and clozapine. Heart rate variability and risk for sudden death in patients with schizophrenia on long-term psychotropic medication. AB - BACKGROUND: Antipsychotic medications cause a wide range of adverse effects and have been associated with sudden death in psychiatric patients. AIMS: To supply power spectral analysis of heart rate variability as a tool to examine the arrythmogenic effects of neuroleptics. METHOD: Heart rate analysis was carried out in patients with schizophrenia on standard doses of neuroleptic monotherapy - 21 were on clozapine, 18 on haloperidol and 17 on olanzapine -- and in 53 healthy subjects. RESULTS: Patients with schizophrenia on clozapine had significantly higher heart rate, lower heart rate variability and lower high frequency and higher low-frequency components compared with patients on haloperidol or olanzapine and matched control subjects. Prolonged QTc intervals were more common in patients than controls. CONCLUSIONS: Patients treated with neuroleptic medications, especially clozapine, showed autonomic dysregulation and cardiac repolarisation changes. Physicians should be aware of this adverse reaction. PMID- 11483481 TI - Increased first-contact rates for very-late-onset schizophrenia-like psychosis in African- and Caribbean-born elders. AB - BACKGROUND: Studies have consistently shown a higher incidence of schizophrenia with onset in early adult life in African and Caribbean migrants to the UK. AIMS: To establish the incidence (first-contact rates) of very-late-onset (>60 years) schizophrenia-like psychosis (SLP) in south London and to test the hypothesis that this is higher among African- and Caribbean-born than indigenous elders. METHOD: We identified all new referrals of SLP to the Maudsley Hospital between 1995 and 2000. Demographic details, including age, ethnicity and electoral ward (address), were obtained from case notes. Incidence was estimated using 1997 census data to determine the denominator population for each ethnic group. RESULTS: The incidence of SLP was significantly higher in African- and Caribbean born than indigenous elders: 172.4 per 100,000 population (95% CI=57.9--286.8) in African- and Caribbean-born males and 323.5 per 100,000 population (95% CI 167.8- 479.1) in African- and Caribbean-born females. Rates also were increased in elders from other immigrant groups, but the numbers involved were too small to reach accepted levels of significance. CONCLUSIONS: Large-scale epidemiological studies are needed to determine both the incidence of and the coexistent risk factors for SLP among all elderly migrants, who may constitute a group with high service needs. PMID- 11483482 TI - Declarations of interest. PMID- 11483483 TI - Comparative cannabis use data. PMID- 11483485 TI - Prognosis of depression and generalised anxiety in primary care. PMID- 11483486 TI - Who is politicising psychiatry in China? PMID- 11483487 TI - The stigma of suicide. PMID- 11483488 TI - Lest we forget, again. PMID- 11483490 TI - An alternative to interruption of treatment in recurrent clozapine-induced severe neutropenia. PMID- 11483492 TI - Cell surface monoamine oxidases: enzymes in search of a function. AB - Ectoenzymes with a catalytically active domain outside the cell surface have the potential to regulate multiple biological processes. A distinct class of copper containing semicarbazide-sensitive monoamine oxidases, expressed on the cell surface and in soluble forms, oxidatively deaminate primary amines. Via transient covalent enzyme-substrate intermediates, this reaction results in production of aldehydes, hydrogen peroxide and ammonium, which are all biologically active substances. The physiological functions of these enzymes have remained unknown, although they have been suggested to be involved in the metabolism of biogenic amines. Recently, new roles have been proposed for these enzymes in regulation of glucose uptake and, even more surprisingly, in leukocyte-endothelial cell interactions. The emerging functions of ectoenzymes in signalling and cell-cell adhesion suggest a novel mode of molecular control of these complex processes. PMID- 11483493 TI - Mycobacterium tuberculosis hemoglobin N displays a protein tunnel suited for O2 diffusion to the heme. AB - Macrophage-generated oxygen- and nitrogen-reactive species control the development of Mycobacterium tuberculosis infection in the host. Mycobacterium tuberculosis 'truncated hemoglobin' N (trHbN) has been related to nitric oxide (NO) detoxification, in response to macrophage nitrosative stress, during the bacterium latent infection stage. The three-dimensional structure of oxygenated trHbN, solved at 1.9 A resolution, displays the two-over-two alpha-helical sandwich fold recently characterized in two homologous truncated hemoglobins, featuring an extra N-terminal alpha-helix and homodimeric assembly. In the absence of a polar distal E7 residue, the O2 heme ligand is stabilized by two hydrogen bonds to TyrB10(33). Strikingly, ligand diffusion to the heme in trHbN may occur via an apolar tunnel/cavity system extending for approximately 28 A through the protein matrix, connecting the heme distal cavity to two distinct protein surface sites. This unique structural feature appears to be conserved in several homologous truncated hemoglobins. It is proposed that in trHbN, heme Fe/O2 stereochemistry and the protein matrix tunnel may promote O2/NO chemistry in vivo, as a M.tuberculosis defense mechanism against macrophage nitrosative stress. PMID- 11483494 TI - Structure of human cystathionine beta-synthase: a unique pyridoxal 5'-phosphate dependent heme protein. AB - Cystathionine beta-synthase (CBS) is a unique heme- containing enzyme that catalyzes a pyridoxal 5'-phosphate (PLP)-dependent condensation of serine and homocysteine to give cystathionine. Deficiency of CBS leads to homocystinuria, an inherited disease of sulfur metabolism characterized by increased levels of the toxic metabolite homocysteine. Here we present the X-ray crystal structure of a truncated form of the enzyme. CBS shares the same fold with O-acetylserine sulfhydrylase but it contains an additional N-terminal heme binding site. This heme binding motif together with a spatially adjacent oxidoreductase active site motif could explain the regulation of its enzyme activity by redox changes. PMID- 11483495 TI - X-ray structure of HPr kinase: a bacterial protein kinase with a P-loop nucleotide-binding domain. AB - HPr kinase/phosphatase (HprK/P) is a key regulatory enzyme controlling carbon metabolism in Gram- positive bacteria. It catalyses the ATP-dependent phosphorylation of Ser46 in HPr, a protein of the phosphotransferase system, and also its dephosphorylation. HprK/P is unrelated to eukaryotic protein kinases, but contains the Walker motif A characteristic of nucleotide-binding proteins. We report here the X-ray structure of an active fragment of Lactobacillus casei HprK/P at 2.8 A resolution, solved by the multiwavelength anomalous dispersion method on a seleniated protein (PDB code 1jb1). The protein is a hexamer, with each subunit containing an ATP-binding domain similar to nucleoside/nucleotide kinases, and a putative HPr-binding domain unrelated to the substrate-binding domains of other kinases. The Walker motif A forms a typical P-loop which binds inorganic phosphate in the crystal. We modelled ATP binding by comparison with adenylate kinase, and designed a tentative model of the complex with HPr based on a docking simulation. The results confirm that HprK/P represents a new family of protein kinases, first identified in bacteria, but which may also have members in eukaryotes. PMID- 11483496 TI - Mutations lowering the phosphatase activity of HPr kinase/phosphatase switch off carbon metabolism. AB - The oligomeric bifunctional HPr kinase/P-Ser-HPr phosphatase (HprK/P) regulates many metabolic functions in Gram-positive bacteria by phosphorylating the phosphocarrier protein HPr at Ser46. We isolated Lactobacillus casei hprK alleles encoding mutant HprK/Ps exhibiting strongly reduced phosphatase, but almost normal kinase activity. Two mutations affected the Walker motif A of HprK/P and four a conserved C-terminal region in contact with the ATP-binding site of an adjacent subunit in the hexamer. Kinase and phosphatase activity appeared to be closely associated and linked to the Walker motif A, but dephosphorylation of seryl-phosphorylated HPr (P-Ser-HPr) is not simply a reversal of the kinase reaction. When the hprKV267F allele was expressed in Bacillus subtilis, the strongly reduced phosphatase activity of the mutant enzyme led to increased amounts of P-Ser-HPr. The hprKV267F mutant was unable to grow on carbohydrates transported by the phosphoenolpyruvate:glycose phosphotransferase system (PTS) and on most non-PTS carbohydrates. Disrupting ccpA relieved the growth defect only on non-PTS sugars, whereas replacing Ser46 in HPr with alanine also restored growth on PTS substrates. PMID- 11483497 TI - Novel modular domain PB1 recognizes PC motif to mediate functional protein protein interactions. AB - Modular domains mediating specific protein-protein interactions play central roles in the formation of complex regulatory networks to execute various cellular activities. Here we identify a novel domain PB1 in the budding yeast protein Bem1p, which functions in polarity establishment, and mammalian p67(phox), which activates the microbicidal phagocyte NADPH oxidase. Each of these specifically recognizes an evolutionarily conserved PC motif to interact directly with Cdc24p (an essential protein for cell polarization) and p40(phox) (a component of the signaling complex for the oxidase), respectively. Swapping the PB1 domain of Bem1p with that of p67(phox), which abolishes its interaction with Cdc24p, confers on cells temperature- sensitive growth and a bilateral mating defect. These phenotypes are suppressed by a mutant Cdc24p harboring the PC motif containing region of p40(phox), which restores the interaction with the altered Bem1p. This domain-swapping experiment demonstrates that Bem1p function requires interaction with Cdc24p, in which the PB1 domain and the PC motif participate as responsible modules. PMID- 11483498 TI - Structure and ligand recognition of the PB1 domain: a novel protein module binding to the PC motif. AB - PB1 domains are novel protein modules capable of binding to target proteins that contain PC motifs. We report here the NMR structure and ligand-binding site of the PB1 domain of the cell polarity establishment protein, Bem1p. In addition, we identify the topology of the PC motif-containing region of Cdc24p by NMR, another cell polarity establishment protein that interacts with Bem1p. The PC motif containing region is a structural domain offering a scaffold to the PC motif. The chemical shift perturbation experiment and the mutagenesis study show that the PC motif is a major structural element that binds to the PB1 domain. A structural database search reveals close similarity between the Bem1p PB1 domain and the c Raf1 Ras-binding domain. However, these domains are functionally distinct from each other. PMID- 11483499 TI - Intracellular re-routing of prion protein prevents propagation of PrP(Sc) and delays onset of prion disease. AB - Prion diseases are fatal and transmissible neurodegenerative disorders linked to an aberrant conformation of the cellular prion protein (PrP(c)). We show that the chemical compound Suramin induced aggregation of PrP in a post-ER/Golgi compartment and prevented further trafficking of PrP(c) to the outer leaflet of the plasma membrane. Instead, misfolded PrP was efficiently re-routed to acidic compartments for intracellular degradation. In contrast to PrP(Sc) in prion infected cells, PrP aggregates formed in the presence of Suramin did not accumulate, were entirely sensitive to proteolytic digestion, had distinct biophysical properties, and were not infectious. The prophylactic potential of Suramin-induced intracellular re-routing was tested in mice. After intraperitoneal infection with scrapie prions, peripheral application of Suramin around the time of inoculation significantly delayed onset of prion disease. Our data reveal a novel quality control mechanism for misfolded PrP isoforms and introduce a new molecular mechanism for anti-prion compounds. PMID- 11483500 TI - Altered behavioral rhythms and clock gene expression in mice with a targeted mutation in the Period1 gene. AB - A group of specialized genes has been defined to govern the molecular mechanisms controlling the circadian clock in mammals. Their expression and the interactions among their products dictate circadian rhythmicity. Three genes homologous to Drosophila period exist in the mouse and are thought to be major players in the biological clock. Here we present the generation of mice in which the founding member of the family, Per1, has been inactivated by homologous recombination. These mice present rhythmicity in locomotor activity, but with a period almost 1 h shorter than wild-type littermates. Moreover, the expression of clock genes in peripheral tissues appears to be delayed in Per1 mutant animals. Importantly, light-induced phase shifting appears conserved. The oscillatory expression of clock genes and the induction of immediate-early genes in response to light in the master clock structure, the suprachiasmatic nucleus, are unaffected. Altogether, these data demonstrate that Per1 plays a distinct role within the Per family, as it may be involved predominantly in peripheral clocks and/or in the output pathways of the circadian clock. PMID- 11483501 TI - P25 and P28 proteins of the malaria ookinete surface have multiple and partially redundant functions. AB - The ookinete surface proteins (P25 and P28) are proven antimalarial transmission blocking vaccine targets, yet their biological functions are unknown. By using single (Sko) and double gene knock-out (Dko) Plasmodium berghei parasites, we show that P25 and P28 share multiple functions during ookinete/oocyst development. In the midgut of mosquitoes, the formation of ookinetes lacking both proteins (Dko parasites) is significantly inhibited due to decreased protection against lethal factors, including protease attack. In addition, Dko ookinetes have a much reduced capacity to traverse the midgut epithelium and to transform into the oocyst stage. P25 and P28 are partially redundant in these functions, since the efficiency of ookinete/oocyst development is only mildly compromised in parasites lacking either P25 or P28 (Sko parasites) compared with that of Dko parasites. The fact that Sko parasites are efficiently transmitted by the mosquito is a compelling reason for including both target antigens in transmission-blocking vaccines. PMID- 11483502 TI - Roles for 147 embryonic lethal genes on C.elegans chromosome I identified by RNA interference and video microscopy. AB - Early embryonic development involves complex events such as the regulation of cell division and the establishment of embryonic polarity. To identify genes involved in these events, we collected four-dimensional time-lapse video recordings of the first three cell divisions and analysed terminal phenotypes after RNA interference of 147 embryonic lethal genes previously identified in a systematic screen of Caenorhabditis elegans chromosome I. Over half gave defects in early processes such as meiosis, the assembly or position of the first mitotic spindle, cytokinesis, and proper nuclear positioning. For some phenotypic classes, the majority of genes are involved in a shared biochemical process. In addition, we identified loss-of-function phenotypes for genes of unknown function, but for which homologues exist in other organisms, shedding light on the function of these uncharacterized genes. When applied to the whole genome, this approach should identify the vast majority of genes required for early cell processes, paving the way for a greatly improved understanding of these processes and their regulation at the molecular level. PMID- 11483503 TI - CHIF, a member of the FXYD protein family, is a regulator of Na,K-ATPase distinct from the gamma-subunit. AB - The biological role of small membrane proteins of the new FXYD family is largely unknown. The best characterized FXYD protein is the gamma-subunit of the Na,K ATPase (NKA) that modulates the Na,K-pump function in the kidney. Here, we report that, similarly to gamma(a) and gamma(b) splice variants, the FXYD protein CHIF (corticosteroid-induced factor) is a type I membrane protein which is associated with NKA in renal tissue, and modulates the Na,K-pump transport when expressed in Xenopus oocytes. In contrast to gamma(a) and gamma(b), which both decrease the apparent Na+ affinity of the Na,K-pump, CHIF significantly increases the Na+ affinity and decreases the apparent K+ affinity due to an increased Na+ competition at external binding sites. The extracytoplasmic FXYD motif is required for stable gamma-subunit and CHIF interaction with NKA, while cytoplasmic, positively charged residues are necessary for the gamma-subunit's association efficiency and for CHIF's functional effects. These data document that CHIF is a new tissue-specific regulator of NKA which probably plays a crucial role in aldosterone-responsive tissues responsible for the maintenance of body Na+ and K+ homeostasis. PMID- 11483504 TI - NEDD8 recruits E2-ubiquitin to SCF E3 ligase. AB - NEDD8/Rub1 is a ubiquitin (Ub)-like post-translational modifier that is covalently linked to cullin (Cul)-family proteins in a manner analogous to ubiquitylation. NEDD8 is known to enhance the ubiquitylating activity of the SCF complex (composed of Skp1, Cul-1, ROC1 and F-box protein), but the mechanistic role is largely unknown. Using an in vitro reconstituted system, we report here that NEDD8 modification of Cul-1 enhances recruitment of Ub-conjugating enzyme Ubc4 (E2) to the SCF complex (E3). This recruitment requires thioester linkage of Ub to Ubc4. Our findings indicate that the NEDD8-modifying system accelerates the formation of the E2-E3 complex, which stimulates protein polyubiquitylation. PMID- 11483505 TI - Dephosphorylation of beta2-syntrophin and Ca2+/mu-calpain-mediated cleavage of ICA512 upon stimulation of insulin secretion. AB - Islet cell autoantigen (ICA) 512 is a receptor-tyrosine phosphatase-like protein associated with the secretory granules of neuroendocrine cells, including pancreatic beta-cells. Binding of its cytoplasmic tail to beta2-syntrophin suggests that ICA512 connects secretory granules to the utrophin complex and the actin cytoskeleton. Here we show that stimulation of insulin secretion from INS-1 cells triggers the biosynthesis of pro-ICA512 and the degradation of its mature form. Inhibition of calpain, which is activated upon stimulation of insulin secretion, prevents the Ca2+-dependent proteolysis of ICA512. In vitro mu-calpain cleaves ICA512 between a putative PEST domain and the beta2-syntrophin binding site, whereas binding of ICA512 to beta2-syntrophin protects the former from cleavage. beta2-syntrophin and its F-actin-binding protein utrophin are enriched in subcellular fractions containing secretory granules. ICA512 preferentially binds phospho-beta2-syntrophin and stimulation of insulin secretion induces the Ca2+-dependent, okadaic acid-sensitive dephosphorylation of beta2-syntrophin. Similarly to calpeptin, okadaic acid inhibits ICA512 proteolysis and insulin secretion. Thus, stimulation of insulin secretion might promote the mobilization of secretory granules by inducing the dissociation of ICA512 from beta2 syntrophin-utrophin complexes and the cleavage of the ICA512 cytoplasmic tail by mu-calpain. PMID- 11483506 TI - Membrane fusion machines of paramyxoviruses: capture of intermediates of fusion. AB - Peptides derived from heptad repeat regions adjacent to the fusion peptide and transmembrane domains of many viral fusion proteins form stable helical bundles and inhibit fusion specifically. Paramyxovirus SV5 fusion (F) protein-mediated fusion and its inhibition by the peptides N-1 and C-1 were analyzed. The temperature dependence of fusion by F suggests that thermal energy, destabilizing proline residues and receptor binding by the hemagglutinin-neuraminidase (HN) protein collectively contribute to F activation from a metastable native state. F mediated fusion was reversibly arrested by low temperature or membrane incorporated lipids, and the resulting F intermediates were characterized. N-1 inhibited an earlier F intermediate than C-1. Co-expression of HN with F lowered the temperature required to attain the N-1-inhibited intermediate, consistent with HN binding to its receptor stimulating a conformational change in F. C-1 bound and inhibited an intermediate of F that could be detected until a point directly preceding membrane merger. The data are consistent with C-1 binding a pre-hairpin intermediate of F and with helical bundle formation being coupled directly to membrane fusion. PMID- 11483507 TI - Ergosterol is required for the Sec18/ATP-dependent priming step of homotypic vacuole fusion. AB - In vitro homotypic fusion of yeast vacuoles occurs in three stages: priming, the Sec18 (NSF)-mediated changes that precede vacuole association; docking, the Ypt7 and SNARE-mediated pairing of vacuoles; and fusion, mediated by calmodulin/V0/t SNARE interactions. Defects in catalysts of each stage result in fragmented (unfused) vacuoles. Strains with deletions in any of ERG genes 3-6, lacking normal ergosterol biosynthesis, have fragmented vacuoles. The ergosterol ligands filipin, nystatin and amphotericin B block the in vitro fusion of vacuoles from wild-type cells. Each of these inhibitors acts at the priming stage to inhibit Sec17p release from vacuoles. A reversible delay in Sec18p action prevents vacuoles from acquiring resistance to any of these three drugs, confirming that their action is on the normal fusion pathway. Ergosterol or cholesterol delivery to wild-type vacuoles stimulates their in vitro fusion, and the in vitro fusion of ergDelta vacuoles requires added sterol. The need for ergosterol for vacuole priming underscores the role of lipids in organizing the membrane elements of this complex reaction. PMID- 11483508 TI - Mammalian Golgi-associated Bicaudal-D2 functions in the dynein-dynactin pathway by interacting with these complexes. AB - Genetic analysis in Drosophila suggests that Bicaudal-D functions in an essential microtubule-based transport pathway, together with cytoplasmic dynein and dynactin. However, the molecular mechanism underlying interactions of these proteins has remained elusive. We show here that a mammalian homologue of Bicaudal-D, BICD2, binds to the dynamitin subunit of dynactin. This interaction is confirmed by mass spectrometry, immunoprecipitation studies and in vitro binding assays. In interphase cells, BICD2 mainly localizes to the Golgi complex and has properties of a peripheral coat protein, yet it also co-localizes with dynactin at microtubule plus ends. Overexpression studies using green fluorescent protein-tagged forms of BICD2 verify its intracellular distribution and co localization with dynactin, and indicate that the C-terminus of BICD2 is responsible for Golgi targeting. Overexpression of the N-terminal domain of BICD2 disrupts minus-end-directed organelle distribution and this portion of BICD2 co precipitates with cytoplasmic dynein. Nocodazole treatment of cells results in an extensive BICD2-dynactin-dynein co-localization. Taken together, these data suggest that mammalian BICD2 plays a role in the dynein- dynactin interaction on the surface of membranous organelles, by associating with these complexes. PMID- 11483509 TI - The p41 isoform of invariant chain is a chaperone for cathepsin L. AB - The p41 splice variant of major histocompatibility complex (MHC) class II associated invariant chain (Ii) contains a 65 aa segment that binds to the active site of cathepsin L (CatL), a lysosomal cysteine protease involved in MHC class II-restricted antigen presentation. This segment is absent from the predominant form of Ii, p31. Here we document the in vivo significance of the p41-CatL interaction. By biochemical means and electron microscopy, we demonstrate that the levels of active CatL are strongly reduced in bone marrow-derived antigen presenting cells that lack p41. This defect mainly concerns the mature two-chain forms of CatL, which depend on p41 to be expressed at wild-type levels. Indeed, pulse-chase analysis suggests that these mature forms of CatL are degraded by endocytic proteases when p41 is absent. We conclude that p41 is required for activity of CatL by stabilizing the mature forms of the enzyme. This suggests that p41 is not merely an inhibitor of CatL enzymatic activity, but serves as a chaperone to help maintain a pool of mature enzyme in late-endocytic compartments of antigen-presenting cells. PMID- 11483510 TI - The 'sequential allosteric ring' mechanism in the eukaryotic chaperonin-assisted folding of actin and tubulin. AB - Folding to completion of actin and tubulin in the eukaryotic cytosol requires their interaction with cytosolic chaperonin CCT [chaperonin containing tailless complex polypeptide 1 (TCP-1)]. Three-dimensional reconstructions of nucleotide free CCT complexed to either actin or tubulin show that CCT stabilizes both cytoskeletal proteins in open and quasi-folded conformations mediated through interactions that are both subunit specific and geometry dependent. Here we find that upon ATP binding, mimicked by the non-hydrolysable analog AMP-PNP (5' adenylyl-imido-diphosphate), to both CCT-alpha-actin and CCT- beta-tubulin complexes, the chaperonin component undergoes concerted movements of the apical domains, resulting in the cavity being closed off by the helical protrusions of the eight apical domains. However, in contrast to the GroE system, generation of this closed state does not induce the release of the substrate into the chaperonin cavity, and both cytoskeletal proteins remain bound to the chaperonin apical domains. Docking of the AMP-PNP-CCT-bound conformations of alpha-actin and beta-tubulin to their respective native atomic structures suggests that both proteins have progressed towards their native states. PMID- 11483511 TI - The Ras-like GTPase Gem is involved in cell shape remodelling and interacts with the novel kinesin-like protein KIF9. AB - Gem belongs to the Rad/Gem/Kir (RGK) subfamily of Ras-related GTPases, which also comprises Rem, Rem2 and Ges. The RGK family members Ges and Rem have been shown to produce endothelial cell sprouting and reorganization of the actin cytoskeleton upon overexpression. Here we show that high intracellular Gem levels promote profound changes in cell morphology and we investigate how this phenotype arises dynamically. We also show that this effect requires intact microtubules and microfilaments, and that Gem is associated with both cytoskeletal components. In order to investigate the mechanisms of Gem recruitment to the cytoskeleton, we performed a yeast two-hybrid screen and identified a novel kinesin-like protein, termed KIF9, as a new Gem interacting partner. We further show that Gem and KIF9 interact by co-immunoprecipitation. Furthermore, Gem and KIF9 display identical patterns of gene expression in different tissues and developmental stages. The Gem- KIF9 interaction reported here is the first molecular link between RGK family members and the microtubule cytoskeleton. PMID- 11483512 TI - PIG-S and PIG-T, essential for GPI anchor attachment to proteins, form a complex with GAA1 and GPI8. AB - Many eukaryotic cell surface proteins are anchored to the plasma membrane via glycosylphosphatidylinositol (GPI). The GPI transamidase mediates GPI anchoring in the endoplasmic reticulum, by replacing a protein's C-terminal GPI attachment signal peptide with a pre-assembled GPI. During this transamidation reaction, the GPI transamidase forms a carbonyl intermediate with a substrate protein. It was known that the GPI transamidase is a complex containing GAA1 and GPI8. Here, we report two new components of this enzyme: PIG-S and PIG-T. To determine roles for PIG-S and PIG-T, we disrupted these genes in mouse F9 cells by homologous recombination. PIG-S and PIG-T knockout cells were defective in transfer of GPI to proteins, particularly in formation of the carbonyl intermediates. We also demonstrate that PIG-S and PIG-T form a protein complex with GAA1 and GPI8, and that PIG-T maintains the complex by stabilizing the expression of GAA1 and GPI8. Saccharomyces cerevisiae Gpi16p (YHR188C) and Gpi17p (YDR434W) are orthologues of PIG-T and PIG-S, respectively. PMID- 11483513 TI - Functional reconstitution of the import of the yeast ADP/ATP carrier mediated by the TIM10 complex. AB - Import of the ADP/ATP carrier (AAC) into mitochondria requires the soluble TIM10 complex to cross the intermembrane space. We report here that Tim9 and Tim10 purified from Escherichia coli can form a complex of the same size as the endogenous complex from yeast mitochondria. This shows that no other mitochondrial protein is required for the formation of the TIM10 complex. Co expression of both proteins rendered Tim9 more soluble and allowed purification of the reconstituted complex in a single step. Urea/EDTA treatment of recombinant Tim10 allowed its import into tim10-ts mitochondria that lack endogenous Tim10 and cannot import AAC. In this way, we were able to (i) reconstitute the TIM10 complex in the intermembrane space and (ii) restore import of AAC to almost wild type levels. The reconstituted TIM10 complex not only facilitated passage of AAC across the outer membrane but also ensured its accurate membrane insertion. We conclude that the TIM10 complex can be formed exclusively from Tim9 and Tim10 and that the reconstituted complex efficiently restores AAC import in a strain lacking the TIM10 complex. PMID- 11483514 TI - Propagation of the apoptotic signal by mitochondrial waves. AB - Generation of mitochondrial signals is believed to be important in the commitment to apoptosis, but the mechanisms coordinating the output of individual mitochondria remain elusive. We show that in cardiac myotubes exposed to apoptotic agents, Ca2+ spikes initiate depolarization of mitochondria in discrete subcellular regions, and these mitochondria initiate slow waves of depolarization and Ca2+ release propagating through the cell. Traveling mitochondrial waves are prevented by Bcl-x(L), involve permeability transition pore (PTP) opening, and yield cytochrome c release, caspase activation and nuclear apoptosis. Mitochondrial Ca2+ uptake is critical for wave propagation, and mitochondria at the origin of waves take up Ca2+ particularly effectively, providing a mechanism that may underlie selection of the initiation sites. Thus, apoptotic agents transform the mitochondria into an excitable state by sensitizing PTP to Ca2+. Expansion of the local excitation by mitochondrial waves propagating through the whole cell can be especially important in activation of the apoptotic machinery in large cells. PMID- 11483515 TI - Protein phosphatase 2A and its B56 regulatory subunit inhibit Wnt signaling in Xenopus. AB - Wnt signaling increases beta-catenin abundance and transcription of Wnt responsive genes. Our previous work suggested that the B56 regulatory subunit of protein phosphatase 2A (PP2A) inhibits Wnt signaling. Okadaic acid (a phosphatase inhibitor) increases, while B56 expression reduces, beta-catenin abundance; B56 also reduces transcription of Wnt-responsive genes. Okadaic acid is a tumor promoter, and the structural A subunit of PP2A is mutated in multiple cancers. Taken together, the evidence suggests that PP2A is a tumor suppressor. However, other studies suggest that PP2A activates Wnt signaling. We now show that the B56, A and catalytic C subunits of PP2A each have ventralizing activity in Xenopus embryos. B56 was epistatically positioned downstream of GSK3beta and axin but upstream of beta-catenin, and axin co-immunoprecipitated B56, A and C subunits, suggesting that PP2A:B56 is in the beta-catenin degradation complex. PP2A appears to be essential for beta-catenin degradation, since beta-catenin degradation was reconstituted in phosphatase-depleted Xenopus egg extracts by PP2A, but not PP1. These results support the hypothesis that PP2A:B56 directly inhibits Wnt signaling and plays a role in development and carcinogenesis. PMID- 11483516 TI - Promoting bone morphogenetic protein signaling through negative regulation of inhibitory Smads. AB - Inhibitory Smads, i.e. Smad6 and Smad7, are potent antagonists of the BMP-Smad pathway by interacting with activated bone morphogenetic protein (BMP) type I receptors and thereby preventing the activation of receptor-regulated Smads, or by competing with activated R-Smads for heteromeric complex formation with Smad4. The molecular mechanisms that underlie the regulation of I-Smad activity have remained elusive. Here we report the identification of a cytoplasmic protein, previously termed associated molecule with the SH3 domain of STAM (AMSH), as a direct binding partner for Smad6. AMSH interacts with Smad6, but not with R- and Co-Smads, upon BMP receptor activation in cultured cells. Consistent with this finding, stimulation of cells with BMP induces a co-localization of Smad6 with AMSH in the cytoplasm. Ectopic expression of AMSH prolongs BMP-induced Smad1 phosphorylation, and potentiates BMP-induced activation of transcriptional reporter activity, growth arrest and apoptosis. The data strongly suggest that the molecular mechanism by which AMSH exerts its action is by inhibiting the binding of Smad6 to activated type I receptors or activated R-Smads. PMID- 11483517 TI - The RING finger protein Siah-1 regulates the level of the transcriptional coactivator OBF-1. AB - The transcriptional coactivator OBF-1, which interacts with Oct-1 and Oct-2 and the octamer site DNA, has been shown to be critical for development of a normal immune response and the formation of germinal centers in secondary lymphoid organs. Here we have identified the RING finger protein Siah-1 as a protein interacting specifically with OBF-1. This interaction is mediated by the C terminal part of Siah-1 and by residues in the N-terminus of OBF-1, partly distinct from the residues required for formation of a complex with the Oct POU domains and the DNA. Interaction between Siah-1 and OBF-1 leads to downregulation of OBF-1 protein level but not mRNA, and to a corresponding reduction in octamer site-dependent transcription activation. Inhibition of the ubiquitin-proteasome pathway in B cells leads to elevated levels of OBF-1 protein. Furthermore, in immunized mice, OBF-1 protein amounts are dramatically increased in primary activated B cells, without concomitant increase in OBF-1 mRNA. These data suggest that Siah-1 is part of a novel regulatory loop controlling the level of OBF-1 protein in B cells. PMID- 11483518 TI - Regulation of BOB.1/OBF.1 stability by SIAH. AB - The BOB.1/OBF.1 coactivator is critically involved in mediating octamer-dependent transcriptional activity in B lymphocytes. Mice lacking this coactivator show various defects in B-cell development, most notably they completely lack germinal centers. Consistent with this phenotype, BOB.1/OBF.1 levels are massively upregulated in germinal center B cells as compared with resting B cells. We have addressed the mechanism of upregulation and found that only a minor part of this regulation can be attributed to increased levels of BOB.1/OBF.1-specific mRNA. Apparently, BOB.1/OBF.1 is also regulated at the protein level. In support of this suggestion we have been able to identify two related BOB.1/OBF.1 interacting proteins, SIAH1 and SIAH2, in a yeast two-hybrid screen. SIAH1 and SIAH2 are known regulators of protein stability. Cotransfection experiments revealed that coexpression of SIAH results in a destabilization of BOB.1/OBF.1 protein without affecting mRNA levels. Further more, proteasome inhibitors block the degradation of BOB.1/OBF.1 protein. Finally, B-cell receptor cross-linking also resulted in the degradation of BOB.1/OBF.1 and consequently reduced transcriptional activation of BOB.1/OBF.1-dependent reporters. PMID- 11483519 TI - Integrity of the N-terminal transcription domain of p53 is required for mutant p53 interference with drug-induced apoptosis. AB - The present study examined whether the ability of mutant p53 to block apoptosis depended on its transcriptional activity. A core domain mutant p53 (143 Val to Ala), in which two N-terminal residues (22 and 23) essential for transactivation were also mutated (Leu to Glu and Trp to Ser, respectively), was examined. While p53 containing only the core mutation efficiently interfered with drug-induced apoptosis, further modification at the N-terminus abolished this blocking activity. Furthermore, expression of c-myc, a suggested target for core mutant p53 transactivation, was elevated in the core mutant p53-expressing cells, but was abolished in the presence of the transcription-deficient p53 core mutant. In addition, wild-type p53, mutated in the N-terminus (residues 22 and 23), was unable to induce apoptosis by itself. Nevertheless, it synergized with drugs in the induction of apoptosis. This suggests that the integrity of the N-terminus is essential for both the activity of wild-type p53 in apoptosis and for mutant p53 mediated block of drug-induced apoptosis. This supports the notion that core p53 mutants act via a gain of function mechanism. PMID- 11483520 TI - Polymerization of the SAM domain of TEL in leukemogenesis and transcriptional repression. AB - TEL is a transcriptional repressor that is a frequent target of chromosomal translocations in a large number of hematalogical malignancies. These rearrangements fuse a potent oligomerization module, the SAM domain of TEL, to a variety of tyrosine kinases or transcriptional regulatory proteins. The self associating property of TEL-SAM is essential for cell transformation in many, if not all of these diseases. Here we show that the TEL-SAM domain forms a helical, head-to-tail polymeric structure held together by strong intermolecular contacts, providing the first clear demonstration that SAM domains can polymerize. Our results also suggest a mechanism by which SAM domains could mediate the spreading of transcriptional repression complexes along the chromosome. PMID- 11483521 TI - An essential nuclear envelope integral membrane protein, Brr6p, required for nuclear transport. AB - Despite rapid advances in our understanding of the function of the nuclear pore complex in nuclear transport, little is known about the role the nuclear envelope itself may play in this critical process. A small number of integral membrane proteins specific to the envelope have been identified in budding yeast, however, none has been reported to affect transport. We have identified an essential gene, BRR6, whose product, Brr6p, behaves like a nuclear envelope integral membrane protein. Notably, the brr6-1 mutant specifically affects transport of mRNA and a protein reporter containing a nuclear export signal. In addition, Brr6p depletion alters nucleoporin distribution and nuclear envelope morphology, suggesting that the protein is required for the spatial organization of nuclear pores. BRR6 interacts genetically with a subset of nucleoporins, and Brr6-green fluorescent protein (GFP) localizes in a punctate nuclear rim pattern, suggesting location at or near the nuclear pore. However, Brr6-GFP fails to redistribute in a (Delta)nup133 mutant, distinguishing Brr6p from known proteins of the pore membrane domain. We hypothesize that Brr6p is located adjacent to the nuclear pore and interacts functionally with the pore and transport machinery. PMID- 11483522 TI - Regulation of alternative pre-mRNA splicing by the ERK MAP-kinase pathway. AB - Differential gene expression through alternative pre-mRNA splicing is crucial to various physiological and pathological conditions. Upon activation of B and T lymphocytes during an immune response, variant isoforms of the cell surface molecule CD44 are generated by alternative pre-mRNA splicing. We show here that in primary mouse T cells as well as in the murine LB-17 T-cell line upregulation of variant CD44 mRNA species upon T-cell activation requires activation of the MEK-ERK pathway. By employing mutant signaling molecules and a novel luciferase based splice reporter system we demonstrate that the Ras-Raf-MEK-ERK signaling cascade, but not the p38 MAP-kinase pathway, activates a mechanism that retains variant CD44 exon v5 sequence in mature mRNA. The findings demonstrate that a highly conserved pleiotropic signaling pathway links extracellular cues to splice regulation, providing an avenue for tissue-specific, developmental or pathology associated splicing decisions. PMID- 11483523 TI - Processing of 20S pre-rRNA to 18S ribosomal RNA in yeast requires Rrp10p, an essential non-ribosomal cytoplasmic protein. AB - Numerous non-ribosomal trans-acting factors involved in pre-ribosomal RNA processing have been characterized, but none of them is specifically required for the last cytoplasmic steps of 18S rRNA maturation. Here we demonstrate that Rio1p/Rrp10p is such a factor. Previous studies showed that the RIO1 gene is essential for cell viability and conserved from archaebacteria to man. We isolated a RIO1 mutant in a screen for mutations synthetically lethal with a mutant allele of GAR1, an essential gene required for 18S rRNA production and rRNA pseudouridylation. We show that RIO1 encodes a cytoplasmic non-ribosomal protein, and that depletion of Rio1p blocks 18S rRNA production leading to 20S pre-rRNA accumulation. In situ hybridization reveals that, in Rio1p depleted cells, 20S pre-rRNA localizes in the cytoplasm, demonstrating that its accumulation is not due to an export defect. This strongly suggests that Rio1p is involved in the cytoplasmic cleavage of 20S pre-rRNA at site D, producing mature 18S rRNA. Thus, Rio1p has been renamed Rrp10p (ribosomal RNA processing #10). Rio1p/Rrp10p is the first non-ribosomal factor characterized specifically required for 20S pre-rRNA processing. PMID- 11483524 TI - The kink-turn: a new RNA secondary structure motif. AB - Analysis of the Haloarcula marismortui large ribosomal subunit has revealed a common RNA structure that we call the kink-turn, or K-turn. The six K-turns in H.marismortui 23S rRNA superimpose with an r.m.s.d. of 1.7 A. There are two K turns in the structure of Thermus thermophilus 16S rRNA, and the structures of U4 snRNA and L30e mRNA fragments form K-turns. The structure has a kink in the phosphodiester backbone that causes a sharp turn in the RNA helix. Its asymmetric internal loop is flanked by C-G base pairs on one side and sheared G-A base pairs on the other, with an A-minor interaction between these two helical stems. A derived consensus secondary structure for the K-turn includes 10 consensus nucleotides out of 15, and predicts its presence in the 5'-UTR of L10 mRNA, helix 78 in Escherichia coli 23S rRNA and human RNase MRP. Five K-turns in 23S rRNA interact with nine proteins. While the observed K-turns interact with proteins of unrelated structures in different ways, they interact with L7Ae and two homologous proteins in the same way. PMID- 11483525 TI - Non-canonical mechanism for translational control in bacteria: synthesis of ribosomal protein S1. AB - Translation initiation region (TIR) of the rpsA mRNA encoding ribosomal protein S1 is one of the most efficient in Escherichia coli despite the absence of a canonical Shine-Dalgarno-element. Its high efficiency is under strong negative autogenous control, a puzzling phenomenon as S1 has no strict sequence specificity. To define sequence and structural elements responsible for translational efficiency and autoregulation of the rpsA mRNA, a series of rpsA' 'lacZ chromosomal fusions bearing various mutations in the rpsA TIR was created and tested for beta-galactosidase activity in the absence and presence of excess S1. These in vivo results, as well as data obtained by in vitro techniques and phylogenetic comparison, allow us to propose a model for the structural and functional organization of the rpsA TIR specific for proteobacteria related to E.coli. According to the model, the high efficiency of translation initiation is provided by a specific fold of the rpsA leader forming a non-contiguous ribosome entry site, which is destroyed upon binding of free S1 when it acts as an autogenous repressor. PMID- 11483526 TI - Truncated initiation factor eIF4G lacking an eIF4E binding site can support capped mRNA translation. AB - Picornavirus proteases cleave translation initiation factor eIF4G into a C terminal two-thirds fragment (hereafter named p100) and an N-terminal one-third fragment, which interacts with the cap-binding factor eIF4E. As the timing of this cleavage correlates broadly with the shut-off of host cell protein synthesis in infected cells, a very widespread presumption has been that p100 cannot support capped mRNA translation. Through the use of an eIF4G-depleted reticulocyte lysate system, we show that this presumption is incorrect. Moreover, recombinant p100 can also reverse the inhibition of capped mRNA translation caused either by m7GpppG cap analogue, by 4E-BP1, which sequesters eIF4E and thus blocks its association with eIF4G, or by cleavage of endogenous eIF4G by picornavirus proteases. The concentration of p100 required for maximum translation of capped mRNAs is approximately 4-fold higher than the endogenous eIF4G concentration in reticulocyte lysates. Our results imply that picornavirus induced shut-off is not due to an intrinsic inability of p100 to support capped mRNA translation, but to the viral RNA outcompeting host cell mRNA for the limiting concentration of p100. PMID- 11483527 TI - Specific cleavage of hyper-edited dsRNAs. AB - Extended double-stranded DNA (dsRNA) duplexes can be hyper-edited by adenosine deaminases that act on RNA (ADARs). Long uninterrupted dsRNA is relatively uncommon in cells, and is frequently associated with infection by DNA or RNA viruses. Moreover, extensive adenosine to inosine editing has been reported for various viruses. A number of cellular antiviral defence strategies are stimulated by dsRNA. An additional mechanism to remove dsRNA from cells may involve hyper editing of dsRNA by ADARs, followed by targeted cleavage. We describe here a cytoplasmic endonuclease activity that specifically cleaves hyper-edited dsRNA. Cleavage occurs at specific sites consisting of alternating IU and UI base pairs. In contrast, unmodified dsRNA and even deaminated dsRNAs that contain four consecutive IU base pairs are not cleaved. Moreover, dsRNAs in which alternating IU and UI base pairs are replaced by isomorphic GU and UG base pairs are not cleaved. Thus, the cleavage of deaminated dsRNA appears to require an RNA structure that is unique to hyper-edited RNA, providing a molecular target for the disposal of hyper-edited viral RNA. PMID- 11483528 TI - Hda, a novel DnaA-related protein, regulates the replication cycle in Escherichia coli. AB - The bacterial DnaA protein binds to the chromosomal origin of replication to trigger a series of initiation reactions, which leads to the loading of DNA polymerase III. In Escherichia coli, once this polymerase initiates DNA synthesis, ATP bound to DnaA is efficiently hydrolyzed to yield the ADP-bound inactivated form. This negative regulation of DnaA, which occurs through interaction with the beta-subunit sliding clamp configuration of the polymerase, functions in the temporal blocking of re-initiation. Here we show that the novel DnaA-related protein, Hda, from E.coli is essential for this regulatory inactivation of DnaA in vitro and in vivo. Our results indicate that the hda gene is required to prevent over-initiation of chromosomal replication and for cell viability. Hda belongs to the chaperone-like ATPase family, AAA(+), as do DnaA and certain eukaryotic proteins essential for the initiation of DNA replication. We propose that the once-per-cell-cycle rule of replication depends on the timely interaction of AAA(+) proteins that comprise the apparatus regulating the activity of the initiator of replication. PMID- 11483529 TI - Stability, chromatin association and functional activity of mammalian pre replication complex proteins during the cell cycle. AB - We have examined the behavior of pre-replication complex (pre-RC) proteins in relation to key cell cycle transitions in Chinese Hamster Ovary (CHO) cells. ORC1, ORC4 and Cdc6 were stable (T1/2 >2 h) and associated with a chromatin containing fraction throughout the cell cycle. Green fluorescent protein-tagged ORC1 associated with chromatin throughout mitosis in living cells and co localized with ORC4 in metaphase spreads. Association of Mcm proteins with chromatin took place during telophase, approximately 30 min after the destruction of geminin and cyclins A and B, and was coincident with the licensing of chromatin to replicate in geminin-supplemented Xenopus egg extracts. Neither Mcm recruitment nor licensing required protein synthesis throughout mitosis. Moreover, licensing could be uncoupled from origin specification in geminin supplemented extracts; site-specific initiation within the dihydrofolate reductase locus required nuclei from cells that had passed through the origin decision point (ODP). These results demonstrate that mammalian pre-RC assembly takes place during telophase, mediated by post-translational modifications of pre existing proteins, and is not sufficient to select specific origin sites. A subsequent, as yet undefined, step selects which pre-RCs will function as replication origins. PMID- 11483530 TI - Excision of deaminated cytosine from the vertebrate genome: role of the SMUG1 uracil-DNA glycosylase. AB - Gene-targeted mice deficient in the evolutionarily conserved uracil-DNA glycosylase encoded by the UNG gene surprisingly lack the mutator phenotype characteristic of bacterial and yeast ung(-) mutants. A complementary uracil-DNA glycosylase activity detected in ung(-/-) murine cells and tissues may be responsible for the repair of deaminated cytosine residues in vivo. Here, specific neutralizing antibodies were used to identify the SMUG1 enzyme as the major uracil-DNA glycosylase in UNG-deficient mice. SMUG1 is present at similar levels in cell nuclei of non-proliferating and proliferating tissues, indicating a replication- independent role in DNA repair. The SMUG1 enzyme is found in vertebrates and insects, whereas it is absent in nematodes, plants and fungi. We propose a model in which SMUG1 has evolved in higher eukaryotes as an anti mutator distinct from the UNG enzyme, the latter being largely localized to replication foci in mammalian cells to counteract de novo dUMP incorporation into DNA. PMID- 11483531 TI - Converting a DNA damage checkpoint effector (UmuD2C) into a lesion bypass polymerase (UmuD'2C). AB - During the SOS response of Escherichia coli to DNA damage, the umuDC operon is induced, producing the trimeric protein complexes UmuD2C, a DNA damage checkpoint effector, and UmuD'2C (DNA polymerase V), which carries out translesion synthesis, the basis of 'SOS mutagenesis'. UmuD'2, the homodimeric component of DNA pol V, is produced from UmuD by RecA-facilitated self-cleavage, which removes the 24 N-terminal residues of UmuD. We report the solution structure of UmuD'2 (PDB ID 1I4V) and interactions within UmuD'-UmuD, a heterodimer inactive in translesion synthesis. The overall shape of UmuD'2 in solution differs substantially from the previously reported crystal structure, even though the topologies of the two structures are quite similar. Most significantly, the active site residues S60 and K97 do not point directly at one another in solution as they do in the crystal, suggesting that self-cleavage of UmuD might require RecA to assemble the active site. Structural differences between UmuD'2 and UmuD' UmuD suggest that UmuD'2C and UmuD2C might achieve their different biological activities through distinct interactions with RecA and DNA pol III. PMID- 11483532 TI - G-overhang dynamics at Tetrahymena telomeres. AB - To learn more about the structure of the DNA terminus at Tetrahymena thermophila telomeres, we have devised a ligation-mediated primer extension protocol to accurately measure the length of the G-strand overhang. We show that overhang length and the identity of the 3'-terminal nucleotide are tightly regulated. The majority of overhangs terminate in the sequence 5'-TTGGGGT and >80% are either 14 15 or 20-21 nucleotides in length. No significant changes in overhang length were detected as cells traversed the cell cycle. However, changes in length distribution were observed when cells exited the cell cycle, indicating an altered balance between DNA synthesis and degradation or end protection. We also provide evidence that rDNA molecules have overhangs on both telomeres. Full length rDNA could be cloned by a strategy that depends on overhangs being present at both ends. Moreover, analysis of leading strand telomeres revealed that a significant fraction have overhangs > or =5 nucleotides. Our results indicate that generation of the terminal telomeric DNA structure is highly regulated and requires several distinct DNA-processing events. PMID- 11483536 TI - Point-counterpoint: should physicians accept gifts from their patients? Yes: If they are given out of beneficence or appreciation. PMID- 11483537 TI - Point-counterpoint: should physicians accept gifts from their patients? No: Gifts debase the true value of care. PMID- 11483533 TI - dim-2 encodes a DNA methyltransferase responsible for all known cytosine methylation in Neurospora. AB - To understand better the control of DNA methylation, we cloned and characterized the dim-2 gene of Neurospora crassa, the only eukaryotic gene currently known in which mutations appear to eliminate DNA methylation. The dim-2 gene is responsible for methylation in both symmetrical and asymmetrical sites. We mapped dim-2 between wc-1 and un-10 on linkage group (LG) VIIR and identified the gene by RFLP mapping and genetic complementation. Dim-2 encodes a 1454 amino acid protein including a C-terminal domain homologous to known DNA methyltransferases (MTases) and a novel N-terminal domain. Neither a deletion that removed the first 186 amino acids of the protein nor a mutation in a putative nucleotide binding site abolished function, but a single amino acid substitution in the predicted catalytic site did. Tests for repeat-induced point mutation (RIP) indicated that dim-2 does not play a role in this process, i.e. duplicated sequences are mutated in dim-2 strains, as usual, but the mutated sequences are not methylated, unlike the situation in dim-2+ strains. We conclude that dim-2 encodes an MTase that is responsible for all DNA methylation in vegetative tissues of NEUROSPORA: PMID- 11483538 TI - Ignorance about our health care workforce: a public health emergency. PMID- 11483539 TI - The evidence house: how to build an inclusive base for complementary medicine. PMID- 11483540 TI - Foreigners flock to Cuba for medical care. PMID- 11483541 TI - Time-release analgesic drug causes fatal overdoses in United States. PMID- 11483543 TI - The risks of being a sheepherder's wife. PMID- 11483545 TI - Spider mythology across the world. PMID- 11483546 TI - Hyponatremic encephalopathy induced by thiazides. PMID- 11483547 TI - Does the aggressive use of polyvalent antivenin for rattlesnake bites result in serious acute side effects? AB - OBJECTIVE: To determine the incidence and severity of acute side effects from the use of polyvalent antivenin in victims of rattlesnake bites. DESIGN: We retrospectively reviewed the records of all patients who presented with rattlesnake bites to a university teaching hospital during an 11-year period. From patient medical records, we extracted demographic data, clinical measurements, and outcomes during emergency department evaluation and subsequent hospitalization. Data regarding serum sickness were not collected. OUTCOME MEASURES: Primary outcome variables were the occurrence of immediate hypersensitivity reaction to antivenin, the type of reaction, permanent disability at hospital discharge, and mortality. RESULTS: We identified a total of 73 patients with rattlesnake bites during the study period. Bite envenomation was graded as nonenvenomated, 7 patients (10%); mild, 23 patients (32%); moderate, 32 patients (44%); and severe, 11 patients (15%). We identified 65 patients who received antivenin. Antivenin doses ranged from 1 to 30 vials per patient (mean, 12.0 +/- 6.0), for a total of 777 vials. In 43 patients (66%), 10 or more vials of antivenin were given. The mean number of vials of antivenin given to each snakebite grade were as follows: mild, 8.4 (+/-4.0); moderate, 11.8 (+/-5.7); and severe, 18.7 (+/-6.3). No deaths, amputations, or permanent disability from snakebite occurred in the patients receiving antivenin. Acute side effects of antivenin-occurring within the first 6 hours after administration were seen in 12 patients (18%; 95% confidence interval, 10%-30%). Acute side effects consisted solely of urticaria in all but 1 patient (2%; 95% confidence interval, 0%-8%). This patient had a history of previous antivenin reaction and required a short course of intravenous epinephrine for blood pressure support. No other complications occurred. CONCLUSION: The administration of polyvalent Crotalidae antivenin is safe. Acute hypersensitivity, when it occurs, consists solely in most cases of urticaria. Serious side effects are uncommon. PMID- 11483548 TI - The antivenin is safe, but its future is uncertain. PMID- 11483549 TI - Personal growth in medical faculty: a qualitative study. AB - BACKGROUND: A physician's effectiveness depends on good communication, and cognitive and technical skills used with wisdom, compassion, and integrity. Attaining the last attributes requires growth in awareness and management of one's feelings, attitudes, beliefs, and life experiences. Yet, little empiric research has been done on physicians' personal growth. OBJECTIVE: To use qualitative methods to understand personal growth in a selected group of medical faculty. DESIGN: Case study, using open-ended survey methods to elicit written descriptions of respondents' personal growth experiences. SETTING: United States and Great Britain. PARTICIPANTS: Facilitators, facilitators-in-training, and members of a personal growth interest group of the American Academy on Physician and Patient, chosen because of their interest, knowledge, and experience in the topic area and their accessibility. MEASUREMENTS: Qualitative analysis of submitted stories included initially identifying and sorting themes, placing themes into categories, applying the categories to the database for verification, and verifying findings by independent reviewers. RESULTS: Of 64 subjects, 32 returned questionnaires containing 42 stories. Respondents and nonrespondents were not significantly different in age, sex, or specialty. The analysis revealed 3 major processes that promoted personal growth: powerful experiences, helping relationships, and introspection. Usually personal growth stories began with a powerful experience or a helping relationship (or both), proceeded to introspection, and ended in a personal growth outcome. Personal growth outcomes included changes in values, goals, or direction; healthier behaviors; improved connectedness with others; improved sense of self; and increased productivity, energy, or creativity. CONCLUSIONS: Powerful experiences, helping relationships, and introspection preceded important personal growth. These findings are consistent with theoretic and empiric adult learning literature and could have implications for medical education and practice. They need to be confirmed in other physician populations. PMID- 11483550 TI - Becoming an emotionally intelligent physician. PMID- 11483551 TI - Seizure disorders: Part 1. Classification and diagnosis. PMID- 11483552 TI - Treating the patient or the population? Part 2. Judging the benefit of a treatment to society as a whole. PMID- 11483553 TI - Nocturnal enuresis. PMID- 11483555 TI - Nickel coinage in the United States: the history of a common contact allergen. PMID- 11483560 TI - Herbal medicine. PMID- 11483557 TI - Hypertensive therapy: attacking the renin-angiotensin system. PMID- 11483561 TI - Medicina Cubana: a fresh perspective. PMID- 11483562 TI - Recognizing animal suffering and death in medicine. PMID- 11483565 TI - Immigrant women's health: nutritional assessment and dietary intervention. PMID- 11483567 TI - Giving and receiving gifts: one perspective. PMID- 11483568 TI - Feudal lords of science and medicine. PMID- 11483570 TI - The search for meaning in noncoding DNA. PMID- 11483571 TI - Transposon-induced hotspots for genomic instability. PMID- 11483572 TI - SAGE thoughts on aging. PMID- 11483573 TI - Gene targeting of Desrt, a novel ARID class DNA-binding protein, causes growth retardation and abnormal development of reproductive organs. AB - We have cloned and characterized a novel murine DNA-binding protein Desrt, with a motif characteristic of the ARID (A-T rich interaction domain) family of transcription factors. The Desrt gene encodes an 83-kD protein that is shown to bind DNA and is widely expressed in adult tissues. To examine the in vivo function of Desrt, we have generated mice with a targeted mutation in the ARID domain of Desrt. Homozygous mutants have reduced viability, pronounced growth retardation, and a high incidence of abnormalities of the female and male reproductive organs including cryptorchidism. This may thus serve as a model to dissect the mechanisms involved in the development of the reproductive tract including testicular descent. Gene-targeted mice also display a reduction in the thickness of the zona reticularis of the adrenal gland and transient aberrations of the T and B cell compartments of primary lymphoid organs. These data show that this novel DNA-binding protein, Desrt, has a nonredundant function during growth and in the development of the reproductive system. PMID- 11483574 TI - Analysis of conserved noncoding DNA in Drosophila reveals similar constraints in intergenic and intronic sequences. AB - Comparative genomic approaches to gene and cis-regulatory prediction are based on the principle that differential DNA sequence conservation reflects variation in functional constraint. Using this principle, we analyze noncoding sequence conservation in Drosophila for 40 loci with known or suspected cis-regulatory function encompassing >100 kb of DNA. We estimate the fraction of noncoding DNA conserved in both intergenic and intronic regions and describe the length distribution of ungapped conserved noncoding blocks. On average, 22%-26% of noncoding sequences surveyed are conserved in Drosophila, with median block length approximately 19 bp. We show that point substitution in conserved noncoding blocks exhibits transition bias as well as lineage effects in base composition, and occurs more than an order of magnitude more frequently than insertion/deletion (indel) substitution. Overall, patterns of noncoding DNA structure and evolution differ remarkably little between intergenic and intronic conserved blocks, suggesting that the effects of transcription per se contribute minimally to the constraints operating on these sequences. The results of this study have implications for the development of alignment and prediction algorithms specific to noncoding DNA, as well as for models of cis-regulatory DNA sequence evolution. PMID- 11483575 TI - Changes in gene expression associated with developmental arrest and longevity in Caenorhabditis elegans. AB - Gene expression in a developmentally arrested, long-lived dauer population of Caenorhabditis elegans was compared with a nondauer (mixed-stage) population by using serial analysis of gene expression (SAGE). Dauer (152,314) and nondauer (148,324) SAGE tags identified 11,130 of the predicted 19,100 C. elegans genes. Genes implicated previously in longevity were expressed abundantly in the dauer library, and new genes potentially important in dauer biology were discovered. Two thousand six hundred eighteen genes were detected only in the nondauer population, whereas 2016 genes were detected only in the dauer, showing that dauer larvae show a surprisingly complex gene expression profile. Evidence for differentially expressed gene transcript isoforms was obtained for 162 genes. H1 histones were differentially expressed, raising the possibility of alternative chromatin packaging. The most abundant tag from dauer larvae (20-fold more abundant than in the nondauer profile) corresponds to a new, unpredicted gene we have named tts-1 (transcribed telomere-like sequence), which may interact with telomeres or telomere-associated proteins. Abundant antisense mitochondrial transcripts (2% of all tags), suggest the existence of an antisense-mediated regulatory mechanism in C. elegans mitochondria. In addition to providing a robust tool for gene expression studies, the SAGE approach already has provided the advantage of new gene/transcript discovery in a metazoan. PMID- 11483576 TI - Molecular characterization of two natural hotspots in the Drosophila buzzatii genome induced by transposon insertions. AB - Transposable elements (TEs) have been implicated in the generation of genetic rearrangements, but their potential to mediate changes in the organization and architecture of host genomes could be even greater than previously thought. Here, we describe the naturally occurring structural and nucleotide variation around two TE insertions in the genome of Drosophila buzzatii. The studied regions correspond to the breakpoints of a widespread chromosomal inversion generated by ectopic recombination between oppositely oriented copies of a TE named Galileo. A detailed molecular analysis by Southern hybridization, PCR amplification, and DNA sequencing of 7.1 kb surrounding the inversion breakpoints in 39 D. buzzatii lines revealed an unprecedented degree of restructuring, consisting of 22 insertions of ten previously undescribed TEs, 13 deletions, 1 duplication, and 1 small inversion. All of these alterations occurred exclusively in inverted chromosomes and appear to have accumulated after the insertion of the Galileo elements, within or close to them. The nucleotide variation at the studied regions is six times lower in inverted than in noninverted chromosomes, suggesting that most of the observed changes originated in only 84,000 years. Galileo elements thus seemed to promote the transformation of these, otherwise normal, chromosomal regions in genetically unstable hotspots and highly efficient traps for transposon insertions. The particular features of two new Galileo copies found indicate that this TE belongs to the Foldback family. Together, our results strengthen the importance of TEs, and especially DNA transposons, as inducers of genome plasticity in evolution. PMID- 11483577 TI - Prokaryotic homologs of the eukaryotic DNA-end-binding protein Ku, novel domains in the Ku protein and prediction of a prokaryotic double-strand break repair system. AB - Homologs of the eukaryotic DNA-end-binding protein Ku were identified in several bacterial and one archeal genome using iterative database searches with sequence profiles. Identification of prokaryotic Ku homologs allowed the dissection of the Ku protein sequences into three distinct domains, the Ku core that is conserved in eukaryotes and prokaryotes, a derived von Willebrand A domain that is fused to the amino terminus of the core in eukaryotic Ku proteins, and the newly recognized helix-extension-helix (HEH) domain that is fused to the carboxyl terminus of the core in eukaryotes and in one of the Ku homologs from the Actinomycete Streptomyces coelicolor. The version of the HEH domain present in eukaryotic Ku proteins represents the previously described DNA-binding domain called SAP. The Ku homolog from S. coelicolor contains a distinct version of the HEH domain that belongs to a previously unnoticed family of nucleic-acid-binding domains, which also includes HEH domains from the bacterial transcription termination factor Rho, bacterial and eukaryotic lysyl-tRNA synthetases, bacteriophage T4 endonuclease VII, and several uncharacterized proteins. The distribution of the Ku homologs in bacteria coincides with that of the archeal eukaryotic-type DNA primase and genes for prokaryotic Ku homologs form predicted operons with genes coding for an ATP-dependent DNA ligase and/or archeal eukaryotic-type DNA primase. Some of these operons additionally encode an uncharacterized protein that may function as nuclease or an Slx1p-like predicted nuclease containing a URI domain. A hypothesis is proposed that the Ku homolog, together with the associated gene products, comprise a previously unrecognized prokaryotic system for repair of double-strand breaks in DNA. PMID- 11483578 TI - Divergence of function in sequence-related groups of Escherichia coli proteins. AB - The most prominent mechanism of molecular evolution is believed to have been duplication and divergence of genes. Proteins that belong to sequence-related groups in any one organism are candidates to have emerged from such a process and to share a common ancestor. Groups of proteins in Escherichia coli having sequence similarity are mostly composed of proteins with closely related function, but some groups comprise proteins with unrelated functions. In order to understand how function can change while sequences remain similar, we have examined some of these groups in detail. The enzymes analyzed in this work include representatives of amidotransferases, phosphotransferases, decarboxylases, and others. Most sequence-related groups contain enzymes that are in the same classes of Enzyme Commission (EC) numbers. We have concentrated on groups that are heterogeneous in that respect, and also on groups containing more than one enzyme of any pathway. We find that although the EC number may differ, the reaction chemistry of these sequence-related proteins is the same or very similar. Some of these families illustrate how diversification has taken place in evolution, using common features of either reaction chemistry or ligand specificity, or both, to create catalysts for different kinds of biochemical reactions. This information has relevance to the area of functional genomics in which the activities of gene products of unknown reading frames are attributed by analogy to the functions of sequence-related proteins of known function. PMID- 11483579 TI - Sequence variation within the fragile X locus. AB - The human genome provides a reference sequence, which is a template for resequencing studies that aim to discover and interpret the record of common ancestry that exists in extant genomes. To understand the nature and pattern of variation and linkage disequilibrium comprising this history, we present a study of approximately 31 kb spanning an approximately 70 kb region of FMR1, sequenced in a sample of 20 humans (worldwide sample) and four great apes (chimp, bonobo, and gorilla). Twenty-five polymorphic sites and two insertion/deletions, distributed in 11 unique haplotypes, were identified among humans. Africans are the only geographic group that do not share any haplotypes with other groups. Parsimony analysis reveals two main clades and suggests that the four major human geographic groups are distributed throughout the phylogenetic tree and within each major clade. An African sample appears to be most closely related to the common ancestor shared with the three other geographic groups. Nucleotide diversity, pi, for this sample is 2.63 +/- 6.28 x 10(-4). The mutation rate, mu is 6.48 x 10(-10) per base pair per year, giving an ancestral population size of approximately 6200 and a time to the most recent common ancestor of approximately 320,000 +/- 72,000 per base pair per year. Linkage disequilibrium (LD) at the FMR1 locus, evaluated by conventional LD analysis and by the length of segment shared between any two chromosomes, is extensive across the region. PMID- 11483580 TI - Gene expression profiling in human fetal liver and identification of tissue- and developmental-stage-specific genes through compiled expression profiles and efficient cloning of full-length cDNAs. AB - Fetal liver intriguingly consists of hepatic parenchymal cells and hematopoietic stem/progenitor cells. Human fetal liver aged 22 wk of gestation (HFL22w) corresponds to the turning point between immigration and emigration of the hematopoietic system. To gain further molecular insight into its developmental and functional characteristics, HFL22w was studied by generating expressed sequence tags (ESTs) and by analyzing the compiled expression profiles of liver at different developmental stages. A total of 13,077 ESTs were sequenced from a 3'-directed cDNA library of HFL22w, and classified as follows: 5819 (44.5%) matched to known genes; 5460 (41.8%) exhibited no significant homology to known genes; and the remaining 1798 (13.7%) were genomic sequences of unknown function, mitochondrial genomic sequences, or repetitive sequences. Integration of ESTs of known human genes generated a profile including 1660 genes that could be divided into 15 gene categories according to their functions. Genes related to general housekeeping, ESTs associated with hematopoiesis, and liver-specific genes were highly expressed. Genes for signal transduction and those associated with diseases, abnormalities, or transcription regulation were also noticeably active. By comparing the expression profiles, we identified six gene groups that were associated with different developmental stages of human fetal liver, tumorigenesis, different physiological functions of Itoh cells against the other types of hepatic cells, and fetal hematopoiesis. The gene expression profile therefore reflected the unique functional characteristics of HFL22w remarkably. Meanwhile, 110 full-length cDNAs of novel genes were cloned and sequenced. These novel genes might contribute to our understanding of the unique functional characteristics of the human fetal liver at 22 wk. PMID- 11483581 TI - Capturing whole-genome characteristics in short sequences using a naive Bayesian classifier. AB - Bacterial genomes have diverged during evolution, resulting in clearcut differences in their nucleotide composition, such as their GC content. The analysis of complete sequences of bacterial genomes also reveals the presence of nonrandom sequence variation, manifest in the frequency profile of specific short oligonucleotides. These frequency profiles constitute highly specific genomic signatures. Based on these differences in oligonucleotide frequency between bacterial genomes, we investigated the possibility of predicting the genome of origin for a specific genomic sequence. To this end, we developed a naive Bayesian classifier and systematically analyzed 28 eubacterial and archaeal genomes. We found that sequences as short as 400 bases could be correctly classified with an accuracy of 85%. We then applied the classifier to the identification of horizontal gene transfer events in whole-genome sequences and demonstrated the validity of our approach by correctly predicting the transfer of both the superoxide dismutase (sodC) and the bioC gene from Haemophilus influenzae to Neisseria meningitis, correctly identifying both the donor and recipient species. We believe that this classification methodology could be a valuable tool in biodiversity studies. PMID- 11483582 TI - Prediction of protein functional domains from sequences using artificial neural networks. AB - An artificial neural network (ANN) solution is described for the recognition of domains in protein sequences. A query sequence is first compared to a reference database of domain sequences by use of and the output data, encoded in the form of six parameters, are forwarded to feed-forward artificial neural networks with six input and six hidden units with sigmoidal transfer function. The recognition is based on the distribution of scores precomputed for the known domain groups in a database versus database comparison. Applications to the prediction of function are discussed. PMID- 11483583 TI - Flexible use of high-density oligonucleotide arrays for single-nucleotide polymorphism discovery and validation. AB - A method for identifying and validating single nucleotide polymorphisms (SNPs) with high-density oligonucleotide arrays without the need for locus-specific polymerase chain reactions (PCR) is described in this report. Genomic DNAs were divided into subsets with complexity of ~10 Mb by restriction enzyme digestion and gel-based fragment size resolution, ligated to a common adaptor, and amplified with one primer in a single PCR reaction. As a demonstration of this approach, a total of 124 SNPs were located in 190 kb of genomic sequences distributed across the entire human genome by hybridizing to high-density variant detection arrays (VDA). A set of independent validation experiments was conducted for these SNPs employing bead-based affinity selection followed by hybridization of the affinity-selected SNP-containing fragments to the same VDA that was used to identify the SNPs. A total of 98.7% (74/75) of these SNPs were confirmed using both DNA dideoxynucleotide sequencing and the VDA methodologies. With flexible sample preparation, high-density oligonucleotide arrays can be tailored for even larger scale genome-wide SNP discovery as well as validation. PMID- 11483584 TI - Creating the gene ontology resource: design and implementation. AB - The exponential growth in the volume of accessible biological information has generated a confusion of voices surrounding the annotation of molecular information about genes and their products. The Gene Ontology (GO) project seeks to provide a set of structured vocabularies for specific biological domains that can be used to describe gene products in any organism. This work includes building three extensive ontologies to describe molecular function, biological process, and cellular component, and providing a community database resource that supports the use of these ontologies. The GO Consortium was initiated by scientists associated with three model organism databases: SGD, the Saccharomyces Genome database; FlyBase, the Drosophila genome database; and MGD/GXD, the Mouse Genome Informatics databases. Additional model organism database groups are joining the project. Each of these model organism information systems is annotating genes and gene products using GO vocabulary terms and incorporating these annotations into their respective model organism databases. Each database contributes its annotation files to a shared GO data resource accessible to the public at http://www.geneontology.org/. The GO site can be used by the community both to recover the GO vocabularies and to access the annotated gene product data sets from the model organism databases. The GO Consortium supports the development of the GO database resource and provides tools enabling curators and researchers to query and manipulate the vocabularies. We believe that the shared development of this molecular annotation resource will contribute to the unification of biological information. PMID- 11483585 TI - A marker-dense physical map of the Bradyrhizobium japonicum genome. AB - Bacterial artificial chromosome (BAC) clones are effective mapping and sequencing reagents for use with a wide variety of small and large genomes. This report describes the development of a physical framework for the genome of Bradyrhizobium japonicum, the nitrogen-fixing symbiont of soybean. A BAC library for B. japonicum was constructed that provides a 77-fold genome coverage based on an estimated genome size of 8.7 Mb. The library contains 4608 clones with an average insert size of 146 kb. To generate a physical map, the entire library was fingerprinted with HindIII, and the fingerprinted clones were assembled into contigs using the software (; Sanger Centre, UK). The analysis placed 3410 clones in six large contigs. The ends of 1152 BAC inserts were sequenced to generate a sequence-tagged connector (STC) framework. To join and orient the contigs, high density BAC colony filters were probed with 41 known gene probes and 17 end sequences from contig boundaries. STC sequences were searched against the public databases using and algorithms. Query results allowed the identification of 113 high probability matches with putative functional identities that were placed on the physical map. Combined with the hybridization data, a high-resolution physical map with 194 positioned markers represented in two large contigs was developed, providing a marker every 45 kb. Of these markers, 177 are known or putative B. japonicum genes. Additionally, 1338 significant results (E < 10(-4)) were manually sorted by function to produce a functionally categorized database of relevant B. japonicum STC sequences that can also be traced to specific locations in the physical map. PMID- 11483586 TI - Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): frequency, length variation, transposon associations, and genetic marker potential. AB - A total of 57.8 Mb of publicly available rice (Oryza sativa L.) DNA sequence was searched to determine the frequency and distribution of different simple sequence repeats (SSRs) in the genome. SSR loci were categorized into two groups based on the length of the repeat motif. Class I, or hypervariable markers, consisted of SSRs > or =20 bp, and Class II, or potentially variable markers, consisted of SSRs > or =12 bp <20 bp. The occurrence of Class I SSRs in end-sequences of EcoRI and HindIII-digested BAC clones was one SSR per 40 Kb, whereas in continuous genomic sequence (represented by 27 fully sequenced BAC and PAC clones), the frequency was one SSR every 16 kb. Class II SSRs were estimated to occur every 3.7 kb in BAC ends and every 1.9 kb in fully sequenced BAC and PAC clones. GC rich trinucleotide repeats (TNRs) were most abundant in protein-coding portions of ESTs and in fully sequenced BACs and PACs, whereas AT-rich TNRs showed no such preference, and di- and tetranucleotide repeats were most frequently found in noncoding, intergenic regions of the rice genome. Microsatellites with poly(AT)n repeats represented the most abundant and polymorphic class of SSRs but were frequently associated with the Micropon family of miniature inverted-repeat transposable elements (MITEs) and were difficult to amplify. A set of 200 Class I SSR markers was developed and integrated into the existing microsatellite map of rice, providing immediate links between the genetic, physical, and sequence-based maps. This contribution brings the number of microsatellite markers that have been rigorously evaluated for amplification, map position, and allelic diversity in Oryza spp. to a total of 500. PMID- 11483587 TI - Virtual genome scan: a tool for restriction landmark-based scanning of the human genome. AB - There is substantial interest in implementing technologies that allow comparisons of whole genomes of individuals and of tissues and cell populations. Restriction landmark genome scanning (RLGS) is a highly resolving gel-based technique in which several thousand fragments in genomic digests are visualized simultaneously and quantitatively analyzed. The widespread use of RLGS has been hampered by difficulty in deriving sequence information for displayed fragments and a lack of whole-genome sequence-based framework for interpreting RLGS patterns. We have developed informatics tools for comparisons of sample derived RLGS patterns with patterns predicted from the human genome sequence and displayed as Virtual Genome Scans (VGS). The tools developed allow sequence prediction of fragments in RLGS patterns obtained with different restriction enzyme combinations. The utility of VGS is demonstrated by the identification of restriction fragment length polymorphisms, and of amplifications, deletions, and methylation changes in tumor derived CpG islands and the characterization of an amplified region in a breast tumor that spanned <230 kb on 17q23. PMID- 11483588 TI - Distinct intramembrane cleavage of the beta-amyloid precursor protein family resembling gamma-secretase-like cleavage of Notch. AB - The intramembrane cleavage of beta-amyloid precursor protein by gamma-secretase is the final step in the generation of amyloid beta-protein. A 59- or 57-residue C-terminal fragment called CTFgamma is produced concomitantly. Putative CTFgamma generated in rat brain membrane preparations was purified and sequenced. Instead of CTFgamma, shorter 50- and 49-residue fragments were identified. In addition, we found similar C-terminal fragments of beta-amyloid precursor-like proteins 1 and 2; these were also cleaved at corresponding sites. This newly identified cleavage occurs at a site two to five residues inside the cytoplasmic membrane boundary, which is very similar to gamma-secretase-like cleavage of Notch 1. PMID- 11483589 TI - The epidermal growth factor receptor regulates interaction of the human DF3/MUC1 carcinoma antigen with c-Src and beta-catenin. AB - The DF3/MUC1 mucin-like, transmembrane glycoprotein is aberrantly overexpressed in most human carcinomas. The MUC1 cytoplasmic domain interacts with the c-Src tyrosine kinase and thereby increases binding of MUC1 and beta-catenin. In the present work, coimmunoprecipitation studies demonstrate that MUC1 associates constitutively with the epidermal growth factor receptor (EGF-R) in human ZR-75-1 breast carcinoma cells. Immunofluorescence studies show that EGF-R and MUC1 associate at the cell membrane. We also show that the activated EGF-R phosphorylates the MUC1 cytoplasmic tail on tyrosine at a YEKV motif that functions as a binding site for the c-Src SH2 domain. The results demonstrate that EGF-R-mediated phosphorylation of MUC1 induces binding of MUC1 to c-Src in cells. Moreover, in vitro and in vivo studies demonstrate that EGF-R increases binding of MUC1 and beta-catenin. These findings support a novel role for EGF-R in regulating interactions of MUC1 with c-Src and beta-catenin. PMID- 11483590 TI - Up-regulation of endothelial cyclooxygenase-2 and prostanoid synthesis by platelets. Role of thromboxane A2. AB - Platelet-vascular endothelial cell interactions are central to the maintenance of vascular homeostasis. Thromboxane A2 (TXA2) and prostacyclin (prostaglandin (PG)I2) are the major products of cyclooxygenase (COX) metabolism by platelets and the vascular endothelium, respectively. Here we report the effects of platelet-endothelial interactions on human umbilical vein endothelial cells (HUVECs) COX-2 expression and prostanoid synthesis. Co-incubation of platelets with HUVECs resulted in a dose-dependent induction in COX-2 expression. This was accompanied by a relatively small increase in thromboxane B2 synthesis (2 ng) by comparison to the production of 6-keto-PGF1alpha and PGE2, which increased by approximately 14 and 12 ng, respectively. Abrogation of platelet-HUVEC interactions excluded direct cell-cell contact as a required event. Preincubation of HUVECs with SQ29548, a TXA2 receptor antagonist, dose-dependently inhibited platelet-induced COX-2 expression and prostanoid synthesis. Similarly, if platelet TXA2 synthesis was inhibited no induction of COX-2 was observed. Furthermore, a TXA2 analog, carbocyclic TXA2, induced HUVEC COX-2 expression and the synthesis of 6-keto-PGF1alpha and PGE2. This was also associated with an increase in the expression and activity of PGI synthase and PGE synthase but not TX synthase. Platelet co-incubation (or TXA2) also selectively activated the p44/42 mitogen-activated protein kinase pathway to regulate HUVEC COX-2 expression. Thus it seems that platelet-derived TXA2 can act in a paracrine manner to up-regulate endothelial COX-2 expression and PGI2 synthesis. These observations are of particular importance given the recent observations regarding selective COX-2 inhibitors and the suppression of PGI2 synthesis. PMID- 11483591 TI - Opposing regulation of choline deficiency-induced apoptosis by p53 and nuclear factor kappaB. AB - We have previously shown that fetal rat brain cells, preneuronal (PC12), and hepatocyte (CWSV-1) cells undergo apoptosis during choline deficiency (CD). The PC12 and epithelial cell culture models were used to determine the molecular mechanism by which CD induces apoptosis. Our data indicate that CD leads to both growth arrest and apoptosis in a subpopulation of cells, which correlate with the up-regulation of the tumor suppressor protein p53 and concurrent up-regulation of the cyclin-dependent kinase-inhibitor p21(WAF1/CIP1). Additionally, CD induced both a G1/S and a G2/M arrest. Transient transfection of a dominant negative p53 (p53DN) construct into PC12 cells, which inhibited endogenous p53 activation, significantly reduced the induction of apoptosis associated with CD. Interestingly, CD also induced the persistent activation of the transcription factor NF-kappaB. Activation of NF-kappaB has been shown to promote cell survival and proposed to antagonize p53. Consistent with this, expression of a super repressor form of IkappaBalpha (SR-IkappaBalpha) that functions to strongly inhibit NF-kappaB activation, profoundly enhanced cell death during CD. In summary, these results suggest that the effects of CD on apoptosis and subsequent cell survival are mediated through two different signaling pathways, p53 and NF kappaB, respectively. Taken together, our data demonstrates the induction of opposing mechanisms associated with nutrient deficiency that may provide a molecular mechanism by which CD promotes carcinogenesis. PMID- 11483592 TI - Protein synthesis-dependent and -independent regulation of hippocampal synapses by brain-derived neurotrophic factor. AB - A fundamental difference between short-term and long-term forms of synaptic plasticity is the dependence on transcription and translation of new genes. Using organotypic cultures of hippocampal slices, we have investigated whether the modulation of synapses by brain-derived neurotrophic factor (BDNF) also requires protein synthesis. Long-term treatment of hippocampal slice cultures with BDNF increased the number of docked vesicles, but not that of reserve pool vesicles, at CA1 excitatory synapses. BDNF also increased the levels of the vesicle proteins synaptophysin, synaptobrevin, and synaptotagmin, without affecting the presynaptic membrane proteins syntaxin and SNAP-25, or the vesicle-binding protein synapsin-I. The increase in synaptophysin and synaptobrevin expression was moderate (2-fold) and occurred within 6 h after BDNF application. In contrast, synaptotagmin expression took 24 h to reach maximum levels (5-fold). The delayed increase in synaptotagmin was blocked by protein synthesis inhibitors, while the early increase in synaptophysin and synaptobrevin was not. Moreover, the BDNF-induced increase of synaptotagmin was blocked by inhibiting the cAMP/protein kinase A (PKA) pathway. However, BDNF did not activate PKA, and application of a PKA activator did not mimic the BDNF effect. Taken together, these results suggest a novel, protein synthesis-dependent form of BDNF modulation that requires cAMP gating. PMID- 11483593 TI - Chronic overexpression of the calcineurin inhibitory gene DSCR1 (Adapt78) is associated with Alzheimer's disease. AB - The DSCR1 (Adapt78) gene was independently discovered as a resident of the "Down syndrome candidate region"and as an "adaptive response"shock or stress gene that is transiently induced during oxidative stress. Recently the DSCR1 (Adapt78) gene product was discovered to be an inhibitor of the serine/threonine phosphatase, calcineurin, and its signaling pathways. We hypothesized that DSCR1 (Adapt78) might also be involved in the development of Alzheimer's disease. To address this question we first studied DSCR1 (Adapt78) in multiple human tissues and found significant expression in brain, spinal cord, kidney, liver, mammary gland, skeletal muscle, and heart. Within the brain DSCR1 (Adapt78) is predominantly expressed in neurons within the cerebral cortex, hippocampus, substantia nigra, thalamus, and medulla oblongata. When we compared DSCR1 (Adapt78) mRNA expression in post-mortem brain samples from Alzheimer's disease patients and individuals who had died with no Alzheimer's diagnosis, we found that DSCR1 (Adapt78) mRNA levels were about twice as high in age-matched Alzheimer's patients as in controls. DSCR1 (Adapt78) mRNA levels were actually three times higher in patients with extensive neurofibrillary tangles (a hallmark of Alzheimer's disease) than in controls. In comparison, post-mortem brain samples from Down syndrome patients (who suffer Alzheimer's symptoms) also exhibited DSCR1 (Adapt78) mRNA levels two to three times higher than controls. Using a cell culture model we discovered that the amyloid beta(1-42) peptide, which is a major component of senile plaques in Alzheimer's, can directly induce increased expression of DSCR1 (Adapt78). Our findings associate DSCR1 (Adapt78) with such major hallmarks of Alzheimer's disease as amyloid protein, senile plaques, and neurofibrillary tangles. PMID- 11483594 TI - Lipid association-induced N- and C-terminal domain reorganization in human apolipoprotein E3. AB - Apolipoprotein E (apoE) is a 299 amino acid, anti-atherogenic protein that plays a key role in regulating plasma lipoprotein metabolism. It is composed of an N terminal (NT) domain (residues 1-191) that is responsible for binding to members of the low density lipoprotein receptor family and a C-terminal (CT) domain (residues 216-299) that anchors the protein to lipoprotein particles by virtue of its high-affinity lipid binding characteristics. Isoform-specific differences in the NT domain that modulate the lipoprotein binding preference elicited by the CT domain suggest the existence and importance of domain interactions in this protein. Employing steady state fluorescence quenching and resonance energy transfer techniques, spatial proximity relationships between the N- and C terminal domains were investigated in recombinant human apoE3. ApoE3 containing a single Trp at position 264 and an N-iodoacetyl-N'-(5-sulfo-1-napthyl) ethylenediamine (AEDANS) moiety covalently attached to the lone Cys residue at position 112 was used (AEDANS-apoE3/W@264). Fluorescence quenching studies revealed a solvent-exposed location for Trp-264. In the lipid-free state, fluorescence resonance energy transfer (FRET) was noted between Trp-264 and AEDANS, with a calculated distance of 27 A between the two fluorophores. Control experiments established that FRET observed in this system is intramolecular. FRET was abolished upon proteolysis in the linker region connecting the NT and CT domains. Lowering the solution pH to 4 induced an increase in the efficiency of intramolecular energy transfer, with the two domains reorienting about 5 A closer to one another. Interdomain FRET was retained in the presence of 0.6-1.0 m guanidine hydrochloride but was lost at higher concentrations, a manifestation of unfolding of the domains and increased distance between the donor-acceptor pair. Interaction of AEDANS-apoE3/W@264 with lipid induced a loss of FRET, attributed to spatial repositioning of the domains by >80 A. The data provide biophysical evidence that, in addition to reported conformational changes in the four-helix bundle configuration induced by lipid association, lipid binding of apoE is accompanied by reorientation of the tertiary disposition of the NT and CT domains. PMID- 11483595 TI - Oxazolidinones mechanism of action: inhibition of the first peptide bond formation. AB - Oxazolidinones are potent inhibitors of bacterial protein biosynthesis. Previous studies have demonstrated that this new class of antimicrobial agent blocks translation by inhibiting initiation complex formation, while post-initiation translation by polysomes and poly(U)-dependent translation is not a target for these compounds. We found that oxazolidinones inhibit translation of natural mRNA templates but have no significant effect on poly(A)-dependent translation. Here we show that various oxazolidinones inhibit ribosomal peptidyltransferase activity in the simple reaction of 70 S ribosomes using initiator-tRNA or N protected CCA-Phe as a P-site substrate and puromycin as an A-site substrate. Steady-state kinetic analysis shows that oxazolidinones display a competitive inhibition pattern with respect to both the P-site and A-site substrates. This is consistent with a rapid equilibrium, ordered mechanism of the peptidyltransferase reaction, wherein binding of the A-site substrate can occur only after complex formation between peptidyltransferase and the P-site substrate. We propose that oxazolidinones inhibit bacterial protein biosynthesis by interfering with the binding of initiator fMet-tRNA(i)(Met) to the ribosomal peptidyltransferase P site, which is vacant only prior to the formation of the first peptide bond. PMID- 11483596 TI - A Ca(2+)-activated NADPH oxidase in testis, spleen, and lymph nodes. AB - Superoxide and its derivatives are increasingly implicated in the regulation of physiological functions from oxygen sensing and blood pressure regulation to lymphocyte activation and sperm-oocyte fusion. Here we describe a novel superoxide-generating NADPH oxidase referred to as NADPH oxidase 5 (NOX5). NOX5 is distantly related to the gp91(phox) subunit of the phagocyte NADPH oxidase with conserved regions crucial for the electron transport (NADPH, FAD and heme binding sites). However, NOX5 has a unique N-terminal extension that contains three EF hand motifs. The mRNA of NOX5 is expressed in pachytene spermatocytes of testis and in B- and T-lymphocyte-rich areas of spleen and lymph nodes. When heterologously expressed, NOX5 was quiescent in unstimulated cells. However, in response to elevations of the cytosolic Ca(2+) concentration it generated large amounts of superoxide. Upon Ca(2+) activation, NOX5 also displayed a second function: it became a proton channel, presumably to compensate charge and pH alterations due to electron export. In summary, we have identified a novel NADPH oxidase that generates superoxide and functions as a H(+) channel in a Ca(2+) dependent manner. NOX5 is likely to be involved in Ca(2+)-activated, redox dependent processes of spermatozoa and lymphocytes such as sperm-oocyte fusion, cell proliferation, and cytokine secretion. PMID- 11483597 TI - SHP1 protein-tyrosine phosphatase inhibits gp91PHOX and p67PHOX expression by inhibiting interaction of PU.1, IRF1, interferon consensus sequence-binding protein, and CREB-binding protein with homologous Cis elements in the CYBB and NCF2 genes. AB - The CYBB and NCF2 genes encode the phagocyte respiratory burst oxidase proteins, gp91PHOX and p67PHOX. Previously, we identified homologous CYBB and NCF2 cis elements that are necessary for lineage-specific transcription during late myeloid differentiation. We determined that these homologous cis elements are activated by PU.1, IRF1, interferon consensus sequence-binding protein (ICSBP), and the CREB-binding protein (CBP). Since expression of PU.1 and ICSBP is lineage restricted, our investigations identified a mechanism of lineage-specific CYBB and NCF2 transcription. Since PU.1, IRF1, ICSBP, and CBP are expressed in undifferentiated myeloid cells, our investigations did not determine the mechanism of differentiation stage-specific CYBB and NCF2 transcription. In the current investigations, we determine that SHP1 protein-tyrosine phosphatase (SHP1 PTP) inhibits gp91PHOX and p67PHOX expression, in undifferentiated myeloid cell lines, by decreasing interaction of PU.1, IRF1, ICSBP, and CBP with the CYBB and NCF2 genes. We also determine that IRF1 and ICSBP are tyrosine-phosphorylated during interferon gamma differentiation of myeloid cell lines, and we identify IRF1 and ICSBP tyrosine residues that are necessary for CYBB and NCF2 transcription. Therefore, these investigations identify a novel mechanism by which SHP1-PTP antagonizes myeloid differentiation and determine that tyrosine phosphorylation of IRF1 and ICSPB mediates stage-specific transcriptional activation in differentiating myeloid cells. PMID- 11483598 TI - The G(2) DNA damage checkpoint delays expression of genes encoding mitotic regulators. AB - Transcriptional control of gene expression contributes to the regulation of diverse cellular processes including cell cycle progression and the cellular response to DNA damage. Global gene expression profiling was performed using p53 deficient human cells to identify genes with G(2)/M-specific and DNA damage responsive expression. Numerous cell cycle-regulated genes were identified, but surprisingly the analysis failed to identify genes activated by ionizing radiation. Instead, significant delays in expression of G(2)/M-specific genes, including known mitotic regulators, were observed following DNA damage. Thus, in the absence of p53, gene induction does not contribute to the G(2) arrest following DNA damage. Rather, the DNA damage checkpoint elicits a G(2) cell cycle arrest, in part, by delaying accumulation of proteins required in mitosis. PMID- 11483599 TI - Mutant p53 cooperates with ETS and selectively up-regulates human MDR1 not MRP1. AB - The most frequently expressed drug resistance genes, MDR1 and MRP1, occur in human tumors with mutant p53. However, it was unknown if mutant p53 transcriptionally regulated both MDR1 and MRP1. We demonstrated that mutant p53 did not activate either the MRP1 promoter or the endogenous gene. In contrast, mutant p53 strongly up-regulated the MDR1 promoter and expression of the endogenous MDR1 gene. Notably, cells that expressed either a transcriptionally inactive mutant p53 or the empty vector showed no endogenous MDR1 up-regulation. Transcriptional activation of the MDR1 promoter by mutant p53 required an Ets binding site, and mutant p53 and Ets-1 synergistically activated MDR1 transcription. Biochemical analysis revealed that Ets-1 interacted exclusively with mutant p53s in vivo but not with wild-type p53. These findings are the first to demonstrate the induction of endogenous MDR1 by mutant p53 and provide insight into the mechanism. PMID- 11483600 TI - Progressive changes in adherens junction structure during intestinal adenoma formation in Apc mutant mice. AB - The C57BL/6J-Min/+ (Min/+) mouse bears a mutant Apc gene and therefore is an important in vivo model of intestinal tumorigenesis. Min/+ mice develop adenomas that exhibit loss of the wild-type Apc allele (Apc(Min/-)). Previously, we found that histologically normal enterocytes bearing a truncated Apc protein (Apc(Min/+)) migrated more slowly in vivo than enterocytes with either wild-type Apc (Apc(+/+)) or with heterozygous loss of Apc protein (Apc(1638N)). To study this phenotype further, we determined the effect of the Apc(Min) mutation upon cell-cell adhesion by examining the components of the adherens junction (AJ). We observed a reduced association between E-cadherin and beta-catenin in Apc(Min/+) enterocytes. Subcellular fractionation of proteins from Apc(+/+), Apc(Min/+), and Apc(Min/-) intestinal tissues revealed a cytoplasmic localization of intact E cadherin only in Apc(Min/+), suggesting E-cadherin internalization in these enterocytes. beta-Catenin tyrosine phosphorylation was also increased in Apc(Min/+) enterocytes, consistent with its dissociation from E-cadherin. Furthermore, Apc(Min/+) enterocytes showed a decreased association between beta catenin and receptor protein-tyrosine phosphatase beta/zeta (RPTPbeta/zeta), and Apc(Min/-) cells demonstrated an association between beta-catenin and receptor protein-tyrosine phosphatase gamma. In contrast to the Apc(Min/+) enterocytes, Apc(Min/-) adenomas displayed increased expression and association of E-cadherin, beta-catenin, and alpha-catenin relative to Apc(+/+) controls. These data show that Apc plays a role in regulating adherens junction structure and function in the intestine. In addition, discovery of these effects in initiated but histologically normal tissue (Apc(Min/+)) defines a pre-adenoma stage of tumorigenesis in the intestinal mucosa. PMID- 11483601 TI - eIF4G functionally differs from eIFiso4G in promoting internal initiation, cap independent translation, and translation of structured mRNAs. AB - Eukaryotic initiation factor (eIF) 4G plays an important role in assembling the initiation complex required for ribosome binding to an mRNA. Plants, animals, and yeast each express two eIF4G homologs, which share only 30, 46, and 53% identity, respectively. We have examined the functional differences between plant eIF4G proteins, referred to as eIF4G and eIFiso4G, when present as subunits of eIF4F and eIFiso4F, respectively. The degree to which a 5'-cap stimulated translation was inversely correlated with the concentration of eIF4F or eIFiso4F and required the poly(A)-binding protein for optimal function. Although eIF4F and eIFiso4F directed translation of unstructured mRNAs, eIF4F supported translation of an mRNA containing 5'-proximal secondary structure substantially better than did eIFiso4F. Moreover, eIF4F stimulated translation from uncapped monocistronic or dicistronic mRNAs to a greater extent than did eIFiso4F. These data suggest that at least some functions of plant eIFiso4F and eIF4F have diverged in that eIFiso4F promotes translation preferentially from unstructured mRNAs, whereas eIF4F can promote translation also from mRNAs that contain a structured 5'-leader and that are uncapped or contain multiple cistrons. This ability may also enable eIF4F to promote translation from standard mRNAs under cellular conditions in which cap-dependent translation is inhibited. PMID- 11483602 TI - AMY-1, a c-Myc-binding protein, is localized in the mitochondria of sperm by association with S-AKAP84, an anchor protein of cAMP-dependent protein kinase. AB - We have reported that a novel c-Myc-binding protein, AMY-1 (associate of Myc-1), stimulated the transcription activity of c-Myc. To access the molecular function of AMY-1, a two-hybrid screening of cDNAs encoding AMY-1-binding proteins was carried out with AMY-1 as a bait using a human HeLa cDNA library, and a clone encoding cAMP-dependent protein kinase anchor protein 149 (AKAP149), was obtained. AMY-1 was found to bind in vitro and in vivo to the regulatory subunit II binding region of AKAP149 and S-AKAP84, a splicing variant of AKAP149 expressed in the testis. AMY-1 was expressed postmeiotically in the testis, as S AKAP84 was expressed. Furthermore, S-AKAP84 and regulatory subunit II, a regulatory subunit of cAMP-dependent protein kinase, made a ternary complex in cells, and AMY-1 was localized in the mitochondria of HeLa and sperm in association with AKAP149 and S-AKAP84, respectively. These results suggest that AMY-1 plays a role in spermatogenesis. PMID- 11483603 TI - Characterization of the human OATP-C (SLC21A6) gene promoter and regulation of liver-specific OATP genes by hepatocyte nuclear factor 1 alpha. AB - OATP-C (SLC21A6) is the predominant Na(+)-independent uptake system for bile salts and bilirubin of human liver and is expressed exclusively at the basolateral (sinusoidal) hepatocyte membrane. To investigate the basis of liver specific expression of OATP-C, we studied promoter function in the two hepatocyte derived cell lines HepG2 and Huh7 and in nonhepatic HeLa cells. OATP-C promoter constructs containing from 66 to 950 nucleotides of 5'-regulatory sequence were active in HepG2 and Huh7 but not HeLa cells, indicating that determinants of hepatocyte-specific expression reside within the minimal promoter. Deoxyribonuclease I footprint analysis revealed a single region that was protected by HepG2 and Huh7 but not HeLa cell nuclear extracts. The liver enriched transcription factor hepatocyte nuclear factor 1 alpha (HNF1 alpha) was shown by mobility shift assays to bind within this footprint. Coexpression of HNF1 alpha stimulated OATP-C promoter activity 30-fold in HepG2 and 49-fold in HeLa cells. Mutation of the HNF1 site abolished promoter function, indicating that HNF1 alpha is critical for hepatocyte-specific OATP-C gene expression. The human OATP8 (SLC21A8) and mouse Oatp4 (Slc21a6) promoters were also responsive to HNF1 alpha coexpression in HepG2 cells. These data support a role for HNF1 alpha as a global regulator of liver-specific bile salt and organic anion transporter genes. PMID- 11483604 TI - A protein kinase associated with apoptosis and tumor suppression: structure, activity, and discovery of peptide substrates. AB - Death-associated protein kinase (DAPK) has been implicated in apoptosis and tumor suppression, depending on cellular conditions, and associated with mechanisms of disease. However, DAPK has not been characterized as an enzyme due to the lack of protein or peptide substrates. Therefore, we determined the structure of DAPK catalytic domain, used a homology model of docked peptide substrate, and synthesized positional scanning substrate libraries in order to discover peptide substrates with K(m) values in the desired 10 microm range and to obtain knowledge about the preferences of DAPK for phosphorylation site sequences. Mutagenesis of DAPK catalytic domain at amino acids conserved among protein kinases or unique to DAPK provided a link between structure and activity. An enzyme assay for DAPK was developed and used to measure activity in adult brain and monitor protein purification based on the physical and chemical properties of the open reading frame of the DAPK cDNA. The results allow insight into substrate preferences and regulation of DAPK, provide a foundation for proteomic investigations and inhibitor discovery, and demonstrate the utility of the experimental approach, which can be extended potentially to kinase open reading frames identified by genome sequencing projects or functional genetics screens and lacking a known substrate. PMID- 11483605 TI - Structure of pyruvate dehydrogenase kinase. Novel folding pattern for a serine protein kinase. AB - The structure of mitochondrial pyruvate dehydrogenase kinase isozyme 2 is of interest because it represents a family of serine-specific protein kinases that lack sequence similarity with all other eukaryotic protein kinases. Similarity exists instead with key motifs of prokaryotic histidine protein kinases and a family of eukaryotic ATPases. The 2.5-A crystal structure reported here reveals that pyruvate dehydrogenase kinase isozyme 2 has two domains of about the same size. The N-terminal half is dominated by a bundle of four amphipathic alpha helices, whereas the C-terminal half is folded into an alpha/beta sandwich that contains the nucleotide-binding site. Analysis of the structure reveals this C terminal domain to be very similar to the nucleotide-binding domain of bacterial histidine kinases, but the catalytic mechanism appears similar to that of the eukaryotic serine kinases and ATPases. PMID- 11483606 TI - Identification of the first trypanosome H/ACA RNA that guides pseudouridine formation on rRNA. AB - In trypanosomes small nucleolar RNA (snoRNA) genes are clustered, and the clusters encode for either single or multiple RNAs. We previously reported on a genomic locus in Leptomonas collosoma that encodes for multiple C/D snoRNAs whose expression is regulated at the processing level (Xu, Y., Liu, L., Lopez-Estrano, C., and Michaeli, S. (2001) J. Biol. Chem. 276, 14289-14298). In this study we have characterized, in the same genomic locus, the first trypanosome H/ACA RNA, which we termed h1. Having a length of 69 nucleotides, h1 has the potential to guide pseudouridylation on 28 S rRNA. The h1 is processed from a long polycistronic transcript that carries both the C/D and h1 snoRNAs. The h1/rRNA duplex obeys the rules for guiding pseudouridylation. Mapping of the pseudouridine site indicated that the predicted U is indeed modified. However, in contrast to all H/ACA RNAs, h1 consists of a single hairpin structure and is the shortest H/ACA RNA described so far. PMID- 11483607 TI - Scurfin (FOXP3) acts as a repressor of transcription and regulates T cell activation. AB - We have recently identified and cloned Foxp3, the gene defective in mice with the scurfy mutation. The immune dysregulation documented in these mice and in humans with mutations in the orthologous gene indicates that the foxp3 gene product, scurfin, is involved in the regulation of T cell activation and differentiation. The autoimmune state observed in these patients with the immune dysregulation polyendocrinopathy, enteropathy, X-linked syndrome, or X-linked autoimmunity allergic dysregulation syndrome also points to a critical role for scurfin in the regulation of T cell homeostasis. FOXP3 encodes a novel member of the forkhead family of transcription factors. Here we demonstrate that this structural domain is required for nuclear localization and DNA binding. Scurfin, transiently expressed in heterologous cells, represses transcription of a reporter containing a multimeric forkhead binding site. Upon overexpression in CD4 T cells, scurfin attenuates activation-induced cytokine production and proliferation. We have identified FKH binding sequences adjacent to critical NFAT regulatory sites in the promoters of several cytokine genes whose expression is sensitive to changes in SFN abundance. Our findings indicate that the ability of scurfin to bind DNA, and presumably repress transcription, plays a paramount role in determining the amplitude of the response of CD4 T cells to activation. PMID- 11483608 TI - BH3 death domain peptide induces cell type-selective mitochondrial outer membrane permeability. AB - The BH3 domain is essential for the release of cytochrome c from mitochondria by pro-apoptotic Bcl-2 family proteins during apoptosis. This study tested the hypothesis that a Bax peptide that includes the BH3 domain can permeabilize the mitochondrial outer membrane and release cytochrome c in the absence of a permeability transition at the mitochondrial inner membrane. BH3 peptide (0.1-60 microm) released cytochrome c from mitochondria in the presence of physiological concentrations of ions in a cell type-selective manner, whereas a BH3 peptide with a single amino acid substitution was ineffective. The release of cytochrome c by BH3 peptide correlated with the presence of endogenous Bax at the mitochondria and its integral membrane insertion. Cytochrome c release was accompanied by adenylate kinase release, was not associated with mitochondrial swelling or substantial loss of electrical potential across the inner membrane, and was unaffected by inhibitors of the permeability transition pore. Cytochrome c release was, however, inhibited by Bcl-2. Although energy-coupled respiration was inhibited after the release of cytochrome c, mitochondria maintained membrane potential in the presence of ATP due to the reversal of the ATP synthase. Overall, results support the hypothesis that BH3 peptide releases cytochrome c by a Bax-dependent process that is independent of the mitochondrial permeability transition pore but regulated by Bcl-2. PMID- 11483609 TI - Tumor suppression by a proapoptotic calcium-activated chloride channel in mammary epithelium. AB - Little is known of the roles played by ion channels in cancer. Here we describe a pair of closely related calcium-activated chloride channels whose differential regulation in normal, apoptotic, and transformed mouse cells suggests that channel function is proapoptotic and antineoplastic. While mCLCA1 predominates over mCLCA2 under normal physiological conditions, this relationship is reversed by apoptotic stress both in developing mammary gland and in cultured HC11 mammary epithelial cells. Consistent with an apoptosis-promoting role, splicing of mCLCA2 is disrupted in apoptosis-resistant tumor cell lines and in HC11 cells selected for resistance to detachment-induced apoptosis (anoikis). Unexpectedly, mCLCA1 message is also down-regulated in these cells by at least 30-fold. These results suggest that both genes antagonize survival of mammary tumor cells by sensitizing them to anoikis. When MCF7 or HEK293 tumor cells were transfected with plasmids encoding either mCLCA1 or mCLCA2, colony formation was greatly reduced relative to a vector-transfected control, demonstrating that calcium-sensitive chloride channel (CLCA) expression is deleterious to tumor cell survival. Furthermore, mammary epithelial cells overexpressing mCLCA2 had twice the rate of apoptosis of normal cells when subjected to serum starvation and formed multinuclear giants at a high frequency in normal culture, suggesting that mCLCA2 can promote either apoptosis or senescence. PMID- 11483610 TI - Ferredoxin:NADP+ oxidoreductase is a subunit of the chloroplast cytochrome b6f complex. AB - Purified detergent-soluble cytochrome b6f complex from chloroplast thylakoid membranes (spinach) and cyanobacteria (Mastigocladus laminosus) was highly active, transferring 300-350 electrons per cyt f/s. Visible absorbance spectra showed a red shift of the cytochrome f alpha-band and the Qy chlorophyll a band in the cyanobacterial complex and an absorbance band in the flavin 450-480-nm region of the chloroplast complex. An additional high molecular weight (M(r) approximately 35,000) polypeptide in the chloroplast complex was seen in SDS polyacrylamide gel electrophoresis at a stoichiometry of approximately 0.9 (cytochrome f)(-1). The extra polypeptide did not stain for heme and was much more accessible to protease than cytochrome f. Electrospray ionization mass spectrometry of CNBr fragments of the 35-kDa polypeptide was diagnostic for ferredoxin:NADP+ oxidoreductase (FNR), as were antibody reactivity to FNR and diaphorase activity. The absence of FNR in the cyanobacterial complex did not impair decyl-plastoquinol-ferricyanide activity. The activity of the FNR in the chloroplast b6f complex was also shown by NADPH reduction, in the presence of added ferredoxin, of 0.8 heme equivalents of the cytochrome b6 subunit. It was inferred that the b6f complex with bound FNR, one equivalent per monomer, provides the membrane protein connection to the main electron transfer chain for ferredoxin-dependent cyclic electron transport. PMID- 11483611 TI - Structure of the tetraspanin main extracellular domain. A partially conserved fold with a structurally variable domain insertion. AB - The tetraspanin family of membrane glycoproteins is involved in the regulation of cellular development, proliferation, activation, and mobility. We have attempted to predict the structural features of the large extracellular domain of tetraspanins (EC2), which is very important in determining their functional specificity. The tetraspanin EC2 is composed of two subdomains: a conserved three helix subdomain and a variable secondary structure subdomain inserted within the conserved subdomain. The occurrence of key disulphide bridges and other invariant residues leads to a conserved relative topology of both subdomains and also suggests a structural classification of tetraspanins. Using the CD81 EC2 structure as a template, the structures of two other EC2s were predicted by homology modeling and indicate a conserved shape, in which the variable subdomain is located at one side of the structure. The conserved and variable subdomains might contain sites that correspond, respectively, to common and specific interactions of tetraspanins. The tetraspanin EC2 seems to correspond to a new scheme of fold conservation/variability among proteins, namely the insertion of a structurally variable subdomain within an otherwise conserved fold. PMID- 11483612 TI - A single gene produces mitochondrial, cytoplasmic, and peroxisomal NADP-dependent isocitrate dehydrogenase in Aspergillus nidulans. AB - NADP-dependent isocitrate dehydrogenase enzymes catalyze the decarboxylation of isocitrate to 2-oxoglutarate accompanied by the production of NADPH. In mammals two different genes encode mitochondrial and cytoplasmic/peroxisomal located enzymes, whereas in Saccharomyces cerevisiae three separate genes specify compartment specific enzymes. We have identified a single gene, idpA, in the filamentous fungus Aspergillus nidulans that specifies a protein with a high degree of identity to mammalian and S. cerevisiae enzymes. Northern blot analysis and reverse transcription-polymerase chain reaction revealed the presence of two idpA transcripts and two transcription start points were identified by sequencing cDNA clones and by 5'-rapid amplification of cDNA ends. The shorter transcript was found to be inducible by acetate and by fatty acids while the longer transcript was present in higher amounts during growth in glucose containing media. The longer transcript is predicted to encode a polypeptide containing an N terminal mitochondrial targeting sequence as well as a C-terminal tripeptide (ARL) as a potential peroxisomal targeting signal. The shorter transcript is predicted to encode a polypeptide lacking the mitochondrial targeting signal but retaining the C-terminal sequence. Immunoblotting using antibody raised against S. cerevisiae Idp1p detected two polypeptides consistent with these predictions. The functions of the predicted targeting sequences were confirmed by microscopic analysis of transformants containing fluorescent protein fusion constructs. Using anti-Idp1p antibodies, protein localization to mitochondria and peroxisomes was observed during growth on glucose whereas cytoplasmic and peroxisomal localization was found upon acetate or fatty acid induction. Therefore, we have established that by the use of two transcription start points a single gene is sufficient to specify localization of NADP-dependent isocitrate dehydrogenase to three different cellular compartments in A. nidulans. PMID- 11483613 TI - Erythropoietin activates two distinct signaling pathways required for the initiation and the elongation of c-myc. AB - Erythropoietin (Epo) stimulation of erythroid cells results in the activation of several kinases and a rapid induction of c-myc expression. Protein kinase C is necessary for Epo up-regulation of c-myc by promoting elongation at the 3'-end of exon 1. PKCepsilon mediates this signal. We now show that Epo triggers two signaling pathways to c-myc. Epo rapidly up-regulated Myc protein in BaF3-EpoR cells. The phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 blocked Myc up regulation in a concentration-dependent manner but had no effect on the Epo induced phosphorylation of ERK1 and ERK2. LY294002 also had no effect on Epo up regulation of c-fos. MEK1 inhibitor PD98059 blocked both the c-myc and the c-fos responses to Epo. PD98059 and the PKC inhibitor H7 also blocked the phosphorylation of ERK1 and ERK2. PD98059 but not LY294002 inhibited Epo induction of ERK1 and ERK2 phosphorylation in normal erythroid cells. LY294002 blocked transcription of c-myc at exon 1. PD98059 had no effect on transcription from exon 1 but, rather, blocked Epo-induced c-myc elongation at the 3'-end of exon 1. These results identify two Epo signaling pathways to c-myc, one of which is PI3K-dependent operating on transcriptional initiation, whereas the other is mitogen-activated protein kinase-dependent operating on elongation. PMID- 11483614 TI - Transcriptional regulation of the p67phox gene: role of AP-1 in concert with myeloid-specific transcription factors. AB - We have investigated the myeloid-specific transcriptional regulation of p67(phox), an essential component of phagocyte respiratory burst NADPH oxidase. Analysis was carried out on the p67(phox) 5'-flanking region from -3669 to -4 (relative to ATG), including the first exon and intron and part of the second exon. The construct extending from -985 to -4 produced the highest luciferase activity in myeloid HL-60 cells but was not active in HeLa or Jurkat cells, indicating myeloid-specific expression. Four active elements were identified: Sp1/Sp3 at -694, PU.1 at -289, AP-1 at -210, and PU.1/HAF1 at -182, the latter three being in the first intron. These cis elements bound their cognate transacting factors both in vitro and in vivo. Mutation of the Sp1, PU.1, or PU.1/HAF1 site each decreased promoter activity by 35-50%. Mutations in all three sites reduced promoter activity by 90%. However, mutation of the AP-1 site alone nearly abolished promoter activity. The AP-1 site bound Jun and Fos proteins from HL-60 cell nuclear extract. Co-expression with Jun B in AP-1-deficient cells increased promoter activity by 3-fold. These data show that full p67(phox) promoter activity requires cooperation between myeloid-specific and nonmyeloid transcription factors, with AP-1 being the most critical for function. PMID- 11483615 TI - The ratio of oxidative phosphorylation complexes I-V in bovine heart mitochondria and the composition of respiratory chain supercomplexes. AB - The ratios of the oxidative phosphorylation complexes NADH:ubiquinone reductase (complex I), succinate:ubiquinone reductase (complex II), ubiquinol:cytochrome c reductase (complex III), cytochrome c oxidase (complex IV), and F1F0-ATP synthase (complex V) from bovine heart mitochondria were determined by applying three novel and independent approaches that gave consistent results: 1) a spectrophotometric-enzymatic assay making use of differential solubilization of complexes II and III and parallel assays of spectra and catalytic activities in the samples before and after ultracentrifugation were used for the determination of the ratios of complexes II, III, and IV; 2) an electrophoretic-densitometric approach using two-dimensional electrophoresis (blue native-polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis) and Coomassie blue staining indices of subunits of complexes was used for determining the ratios of complexes I, III, IV, and V; and 3) two electrophoretic-densitometric approaches that are independent of the use of staining indices were used for determining the ratio of complexes I and III. For complexes I, II, III, IV, and V in bovine heart mitochondria, a ratio 1.1 +/- 0.2:1.3 +/- 0.1:3:6.7 +/- 0.8:3.5 +/- 0.2 was determined. PMID- 11483616 TI - Identification of a class of small molecule inhibitors of the sirtuin family of NAD-dependent deacetylases by phenotypic screening. AB - The yeast transcriptional repressor Sir2p silences gene expression from the telomeric, rDNA, and silent mating-type loci and may play a role in higher order processes such as aging. Sir2p is the founding member of a large family of NAD dependent deacetylase enzymes, named the sirtuins. These proteins are conserved from prokaryotes to eukaryotes, but most remain uncharacterized, including all seven human sirtuins. A reverse chemical genetic approach would be useful in identifying the biological function of sirtuins in a wide variety of experimental systems, but no cell-permeable small molecule inhibitors of sirtuins have been reported previously. Herein we describe a high throughput, phenotypic screen in cells that led to the discovery of a class of sirtuin inhibitors. All three compounds inhibited yeast Sir2p transcriptional silencing activity in vivo, and yeast Sir2p and human SIRT2 deacetylase activity in vitro. Such specific results demonstrate the utility and robustness of this screening methodology. Structure activity relationship analysis of the compounds identified a key hydroxy napthaldehyde moiety that is necessary and sufficient for inhibitory activity. Preliminary studies using one of these compounds suggest that inhibition of sirtuins interferes with body axis formation in Arabidopsis. PMID- 11483617 TI - ABCA1. The gatekeeper for eliminating excess tissue cholesterol. AB - It is widely believed that HDL functions to transport cholesterol from peripheral cells to the liver by reverse cholesterol transport, a pathway that may protect against atherosclerosis by clearing excess cholesterol from arterial cells. A cellular ATP-binding cassette transporter (ABC) called ABCA1 mediates the first step of reverse cholesterol transport: the transfer of cellular cholesterol and phospholipids to lipid-poor apolipoproteins. Mutations in ABCA1 cause Tangier disease (TD), a severe HDL deficiency syndrome characterized by accumulation of cholesterol in tissue macrophages and prevalent atherosclerosis. Studies of TD heterozygotes revealed that ABCA1 activity is a major determinant of plasma HDL levels and susceptibility to CVD. Drugs that induce ABCA1 in mice increase clearance of cholesterol from tissues and inhibit intestinal absorption of dietary cholesterol. Multiple factors related to lipid metabolism and other processes modulate expression and tissue distribution of ABCA1.Therefore, as the primary gatekeeper for eliminating tissue cholesterol, ABCA1 has a major impact on cellular and whole body cholesterol metabolism and is likely to play an important role in protecting against cardiovascular disease. PMID- 11483618 TI - Novel site in lipoprotein lipase (LPL415;-438) essential for substrate interaction and dimer stability. AB - LPL, like other lipases, has the ability to hydrolyze water-insoluble lipid substrates, but the mechanism is incompletely understood. We previously demonstrated a 22-amino acid loop in the amino-terminal domain of LPL to be essential for interaction with lipid substrates (Dugi, K. A., H. L. Dichek, G. D. Talley, H. B. Brewer, Jr., and S. Santamarina-Fojo. 1992. J. Biol. Chem. 267: 25086-25091) and mediation of substrate specificity (Dugi, K. A., H. L. Dichek, and S. Santamarina-Fojo. 1995. J. Biol. Chem. 270: 25396-25401). The carboxy terminal domain, LPL415-438, contains two highly conserved hydrophobic stretches, and represents a candidate region for substrate interactions. Specific point mutations or deletion of the region between the hydrophobic stretches (LPL419 430) caused up to 90% selective loss of hydrolyzing activity against water insoluble triolein, but not against water-soluble tributyrin, implicating a crucial function for LPL419-430 in the interaction with lipid substrates. In contrast, mutations introduced into the hydrophobic regions led to concomitant changes in tributyrin and triolein activities. The presence of an additional positive charge at position 416 yielded a gain of function mutant with 3-fold increased activity. This mutant was about three times more stable at 37 degrees C than wild-type LPL, suggesting an important role for the hydrophobic regions in LPL dimer stability. In summary, our data demonstrate that the carboxy-terminal region LPL415-438 plays an important role in both the interaction of LPL with lipid substrates and the stability of the LPL homodimer. PMID- 11483619 TI - Immunochemical detection of a lipofuscin-like fluorophore derived from malondialdehyde and lysine. AB - The accumulation of fluorescent age pigment or lipofuscin is a frequently observed age-associated cellular alteration in a variety of postmitotic cells of many species. These pigments are observed within granules composed, in part, of damaged protein and lipid. Modification of various biomolecules by aldehyde products of lipid peroxidation is believed to contribute to lipofuscin and ceroid formation. In the present study, we raised a monoclonal antibody (MAb 1F83) directed to the malondialdehyde-modified protein and identified a lipofuscin-like dihydropyridine fluorophore as the major epitope. This antibody was used to conclusively demonstrate that the fluorophore forms on oxidatively modified low density lipoproteins. In addition, we demonstrated that the materials immunoreactive to MAb 1F83 indeed constituted the atherosclerotic lesions, in which intense positivity was associated primarily with macrophage-derived foam cells. The results of this study suggest that the reaction between the lipid peroxidation-derived aldehyde and primary amino groups of protein might represent a process common to the formation of the lipofuscin-like fluorophore during aging and its related diseases. PMID- 11483620 TI - Apoptosis of Neuro2a cells induced by lysosphingolipids with naturally occurring stereochemical configurations. AB - Lysosphingolipids, which lack the fatty acid moiety of sphingolipids, are known to be accumulated in some variants of sphingolipid storage diseases. Here, we report that lysosphingolipids with naturally occurring stereochemical configurations induce apoptosis in mouse neuroblastoma Neuro2a cells. The intracellular dehydrogenase activity and [3H]thymidine incorporation of Neuro2a cells were strongly suppressed by the addition of lysosphingolipids in a dose dependent manner, whereas the parental sphingolipids had no effect. Intranucleosomal DNA fragmentation, chromatin condensation, and phosphatidylserine externalization, which are typical features of apoptosis, were observed when the cells were cultured with 40-80 microM of lysosphingolipids for 24-48 h in the presence of 5% fetal calf serum. Activation of caspase-3-like enzyme occurred after addition of lysosphingolipids followed by incubation at 37 degrees C for 24 h. The addition of an inhibitor of caspases, ZVAD-fmk, to the Neuro2a cell culture completely inhibited the elevation of caspase-3 activity but not the DNA fragmentation. These results may indicate that a caspase-3 independent signaling pathway is involved in the lysosphingolipid-induced apoptosis and suggest that accumulation of lysosphingolipids, but not parental sphingolipids, triggers the apoptotic cascade in neuronal cells of patients with sphingolipidoses. PMID- 11483621 TI - The OSBP-related protein family in humans. AB - Oxysterols are oxygenated derivatives of cholesterol that have a number of biological effects and play a key role in the maintenance of the body cholesterol balance. In this study, we describe the cDNA sequences and genomic structures of the recently identified human oxysterol-binding protein (OSBP)-related protein (ORP) family (Laitinen, S. et al. 1999. J. Lipid Res. 40: 2204-2211). The family now includes 12 genes/proteins, which can be divided into six distinct subfamilies. The ORP have two major structural features: a highly conserved OSBP type sterol-binding domain in the C-terminal half and a pleckstrin homology domain present in the N-terminal region of most family members. Several ORP genes are present in S. cerevisiae, D. melanogaster, and C. elegans, suggesting that the protein family has functions of fundamental importance in the eukaryotic kingdom. Analysis of ORP mRNA levels in unloaded or acetylated LDL-loaded human macrophages revealed that the expression of ORP genes was not significantly affected by the loading, with the exception of ORP6, which was up-regulated 2 fold. The present study summarizes the basic characteristics of the OSBP-related gene/protein family in humans, and provides tools for functional analysis of the encoded proteins. PMID- 11483622 TI - Inhibitory action of gemfibrozil on cholesterol absorption in rat intestine. AB - This study was designed to determine whether gemfibrozil inhibits intestinal lipid absorption. Male Sprague-Dawley rats received an oral dose of 30 mg gemfibrozil/kg body weight for 14 days. Mesenteric lymph cannulation was performed, and a lipid infusion containing 40 micromol/h (35.4 mg/h) of radiolabeled triolein and 2.74 micromol/h (1.06 mg/h) of radiolabeled cholesterol with the addition of 1 mg/h of gemfibrozil was infused intraduodenally at a rate of 3 ml/h for 8 h. The lymph was collected, and the radioactivity levels of the lumen and gut mucosa were measured after the infusion. Lymph cholesterol transport was depressed in gemfibrozil-treated rats, in terms of mass measurements as well as radioactivity in a lesser degree. More radioactive cholesterol remained in the proximal portion of the intestinal lumen and mucosa in the treated rats than in the control rats. More radioactive triglycerides also remained in the proximal intestinal lumen of treated rats, although no difference in lymphatic triglyceride transport was observed between the groups. A significant portion of the radioactive cholesterol remained in the lumen in the gemfibrozil-treated rats. Gemfibrozil increased biliary cholesterol excretion. Thus, this study shows that gemfibrozil inhibits cholesterol absorption in rat intestine. PMID- 11483623 TI - Effects of arachidonic and docosahexaenoic acids on secretion and degradation of bile salt-dependent lipase in AR4-2J cells. AB - In this study we demonstrated that two polyunsaturated fatty acids, arachidonic acid (AA, n-6) and docosahexaenoic acid (DHA, n-3), modulate the secretion of bile salt-dependent lipase (BSDL) by pancreatic AR4-2J cells. The effects of AA and DHA were also compared with that of the monounsaturated fatty acid, oleic acid (OA). Our results showed that the chronic treatment of cells with AA or DHA, that did not affect the biosynthesis rate of BSDL, similarly decreased the amount of secreted BSDL and perturbed the intracellular partitioning of the enzyme, whereas OA had no effect. Particularly, AA and DHA induced the retention of the enzyme in microsomes and lowered its content in the cell cytosol. We have further shown that AA treatment decreased the ubiquitination of the protein, and consequently diminished its export toward the cytosol, a result that might explain the retention of BSDL in microsomes and correlated with membrane phospholipids alteration. The retained protein was further degraded by a nonproteasomal pathway that likely involves ATP-dependent endoplasmic reticulum proteases. These findings concerning the regulation of the pancreatic BSDL secretion by two polyunsaturated acids, AA and DHA, might be of physiological importance in the plasmatic and cellular cholesterol homeostasis. PMID- 11483624 TI - CD36 does not play a direct role in HDL or LDL metabolism. AB - CD36 and scavenger receptor class B, type I (SR-BI) are both class B scavenger receptors that recognize a broad variety of ligands, including oxidized low density lipoprotein (oxLDL), HDL, anionic phospholipids, and apoptotic cells. In this study we investigated the role of mouse CD36 (mCD36) as a physiological lipoprotein receptor. We compared the association of various lipoprotein particles with mCD36 and mSR-BI expressed in COS cells by adenovirus-mediated gene transfer. mCD36 bound human oxLDL and mouse HDL with high affinity. Human LDL bound poorly to mCD36, indicating that mCD36 is unlikely to play a significant role in LDL metabolism. The ability of mCD36 to mediate the selective uptake of cholesteryl esters (CE) from receptor-bound HDL was assessed. In comparison with mSR-BI, mCD36 inefficiently mediated the selective uptake of CE. Hepatic overexpression of mCD36 in C57BL/6 mice by adenovirus-mediated gene transfer did not result in significant alterations in plasma LDL and HDL levels. We conclude that mCD36, while able to bind HDL with high affinity, does not contribute significantly to HDL or LDL metabolism. PMID- 11483625 TI - Distinct patterns of lipoproteins with apoB defined by presence of apoE or apoC III in hypercholesterolemia and hypertriglyceridemia. AB - Apolipoprotein (apo) E and apoC-III concentrations in VLDL and LDL are associated with coronary heart disease. We studied the relationship between apoE and apoC III and the abnormal concentrations and distribution of apoB lipoproteins in 10 hypercholesterolemic and 13 hypertriglyceridemic patients compared with 12 normolipidemic subjects (mean age, 45 years). Sixteen distinct types of apoB lipoprotein particles were separated by first using anti-apoE and anti-apoC-III immunoaffinity chromatography in sequence and then ultracentrifugation [light VLDL, dense VLDL, IDL, and LDL, with apoE with or without apoC-III (E(+)C-III(+), E(+)C-III(-)) or without apoE with or without apoC-III (E(-)C-III(+), E(-)C-III( ))]. The concentrations of VLDL particles with apoC-III (E(+)C-III(+), E(-)C III(+)) were increased in the hypertriglyceridemic group compared with the hypercholesterolemic and normolipidemic groups. These particles were the most triglyceride rich of the particle types, and their triglyceride content was twice as high in hypertriglyceridemics compared with the other two groups. Hypertriglyceridemics had a similar concentration of total E(-)C-III(-) particles compared with normolipidemics, but the E(-)C-III(-) particles were distributed more to VLDL and IDL than to LDL. Hypercholesterolemics, in contrast, were distinguished from the normolipidemic group by 2-fold higher concentrations of apoB lipoproteins without apoE or apoC-III (E(-)C-III(-)), mainly LDL, which had high cholesterol content. Nonetheless, both normolipidemics and hypercholesterolemics had apoC-III-containing VLDL, which comprised 68% and 43% of their total VLDL particles. E(+)C-III(-) particles were a minor type, comprising <10% of particles in all lipoproteins and patient groups. Therefore, VLDL particles with apoC-III may play a central role in identifying the high risk of coronary heart disease in hypertriglyceridemia, but their substantial prevalence in normolipidemics may be of clinical significance as well. PMID- 11483626 TI - Hyodeoxycholic acid efficiently suppresses atherosclerosis formation and plasma cholesterol levels in mice. AB - We examined the effect of hyodeoxycholic acid (HDCA) on plasma cholesterol levels and atherosclerosis in mice. In wild-type C57BL/6 mice, feeding increasing amounts of HDCA resulted in i) progressive decrease in dietary cholesterol absorption, ii) increased concentrations of HDCA in the gallbladder bile, iii) decreased liver cholesterol content, iv) increased liver cholesterol synthesis, and v) increased plasma concentrations of HDCA. In C57BL/6 LDL-receptor knockouts (LDLR-KO) the addition of HDCA to chow and a 0.5% cholesterol diet decreased their total plasma cholesterol levels by 21% and 62%, respectively, because of a decrease in VLDL and LDL cholesterol. Turnover studies showed that HDCA has no effect on VLDL removal from plasma. Furthermore, the addition of HDCA to chow- and 0.5% cholesterol-fed LDLR-KO mice decreased the aortic root atherosclerosis lesion area by 50% and 80%, respectively. Finally, we tested the effect of HDCA on intestinal tumor formation. Feeding C57BL/6 ApcMin mice with HDCA did not affect the number of tumors but decreased the tumor volume in these animals. These results suggest that HDCA might have beneficial effects in the treatment of increased plasma cholesterol levels and atherosclerosis. PMID- 11483627 TI - Physiological compartmental analysis of alpha-linolenic acid metabolism in adult humans. AB - A physiological compartmental model of alpha-linolenic acid metabolism was derived from the plasma concentration-time curves for d5-18:3n-3, d5-20:5n-3, d5 22:5n-3, and d5-22:6n-3 in eight healthy subjects. Subjects received a 1-g oral dose of an isotope tracer of alpha-linolenate (d5-18:3n-3 ethyl ester) while subsisting on a rigorously controlled beef-based diet. By utilizing the Windows Simulation and Analysis Modeling program, kinetic parameters were determined for each subject. Half-lives and mean transit times of the n-3 fatty acids in the plasma were also determined. The model predicted plasma values for the n-3 fatty acids in good accordance with the measured steady state concentrations and also predicted dietary linolenic acid intake for each subject in accordance with values determined by lipid analysis of the diet. Only about 0.2% of the plasma 18:3n-3 was destined for synthesis of 20:5n-3, approximately 63% of the plasma 20:5n-3 was accessible for production of 22:5n-3, and 37% of 22:5n-3 was available for synthesis of 22:6n-3. The inefficiency of the conversion of 18:3n-3 to 20:5n-3 indicates that the biosynthesis of long-chain n-3 PUFA from alpha linolenic acid is limited in healthy individuals. In contrast, the much greater rate of transfer of mass from the plasma 20:5n-3 compartment to 22:5n-3 suggests that dietary eicosapentaenoic acid may be well utilized in the biosynthesis of 22:6n-3 in humans. PMID- 11483628 TI - CTP:phosphocholine cytidylyltransferase, a new sterol- and SREBP-responsive gene. AB - The CTP:phosphocholine cytidylyltransferase (CT) gene encodes the rate controlling enzyme in the phosphatidylcholine biosynthesis pathway. CTalpha mRNA levels, like farnesyl diphosphate synthase and the LDL receptor, are repressed when human or rodent cells are incubated with exogenous sterols and induced when cells are incubated in lipid-depleted medium. A putative sterol response element (SRE) was identified 156 bp upstream of the transcription start site of the CTalpha gene. Electrophoretic mobility shift assays demonstrate that recombinant SREBP-1a binds to the wild-type SRE identified in the CTalpha promoter but not to oligonucleotides containing two mutations in the SRE. In other studies, a luciferase reporter construct under the control of the murine CTalpha proximal promoter was transiently transfected into cells. The activity of the reporter was repressed after addition of sterols to the medium and induced when the cells were incubated in lipid-depleted medium. The activity of the CTalpha-luciferase reporter was also induced when cells were cotransfected with plasmids encoding either SREBP-1a or SREBP-2. In contrast, no induction was observed under the same conditions when the CTalpha promoter-reporter gene contained two mutations in the SRE. In addition, the induction of the wild-type CTalpha promoter-reporter gene that occurs in cells incubated in lipid-depleted medium is attenuated when dominant-negative SREBP is cotransfected into the cells. These studies demonstrate that transcription of the CTalpha gene is inhibited by sterols and activated by mature forms of SREBP. We conclude that SREBP-regulated genes are involved not only in the synthesis of cholesterol, fatty acids, triglycerides, and NADPH, but also, as shown here, in the synthesis of phospholipids. PMID- 11483629 TI - Cholesterol crystallization in model biles: effects of bile salt and phospholipid species composition. AB - Cholesterol in human bile is solubilized in micelles by (relatively hydrophobic) bile salts and phosphatidylcholine (unsaturated acyl chains at sn-2 position). Hydrophilic tauroursodeoxycholate, dipalmitoyl phosphatidylcholine, and sphingomyelin all decrease cholesterol crystal-containing zones in the equilibrium ternary phase diagram (van Erpecum, K. J., and M. C. Carey. 1997. Biochim. Biophys. Acta. 1345: 269-282) and thus could be valuable in gallstone prevention. We have now compared crystallization in cholesterol-supersaturated model systems (3.6 g/dl, 37 degrees C) composed of various bile salts as well as egg yolk phosphatidylcholine (unsaturated acyl chains at sn-2 position), dipalmitoyl phosphatidylcholine, or sphingomyelin throughout the phase diagram. At low phospholipid contents [left two-phase (micelle plus crystal-containing) zone], tauroursodeoxycholate, dipalmitoyl phosphatidylcholine, and sphingomyelin all enhanced crystallization. At pathophysiologically relevant intermediate phospholipid contents [central three-phase (micelle plus vesicle plus crystal containing) zone], tauroursodeoxycholate inhibited, but dipalmitoyl phosphatidylcholine and sphingomyelin enhanced, crystallization. Also, during 10 days of incubation, there was a strong decrease in vesicular cholesterol contents and vesicular cholesterol-to-phospholipid ratios (approximately 1 on day 10), coinciding with a strong increase in crystal mass. At high phospholipid contents [right two-phase (micelle plus vesicle-containing) zone], vesicles were always unsaturated and crystallization did not occur. Strategies aiming to increase amounts of hydrophilic bile salts may be preferable to increasing saturated phospholipids in bile, because the latter may enhance crystallization. PMID- 11483630 TI - Identification of potential substrate-binding sites in yeast and human acyl-CoA sterol acyltransferases by mutagenesis of conserved sequences. AB - In mammals, the esterification of sterols by ACAT plays a critical role in eukaryotic lipid homeostasis. Using the predominant isoform of the yeast ACAT related enzyme family, Are2p, as a model, we targeted phylogenetically conserved sequences for mutagenesis in order to identify functionally important motifs. Deletion, truncation, and missense mutations implicate a regulatory role for the amino-terminal domain of Are2p and identified two carboxyl-terminal motifs as required for catalytic activity. A serine-to-leucine mutation in the (H/Y)SF motif (residues 338-340), unique to sterol esterification enzymes, nullified the activity and stability of yeast Are2p. Similarly, a tyrosine-to-alanine change in the FYxDWWN motif of Are2p (residues 523-529) produced an enzyme with decreased activity and apparent affinity for oleoyl-CoA. Mutagenesis of the tryptophan residues in this motif completely abolished activity. In human ACAT1, mutagenesis of the corresponding motifs (residues 268-270, and 403-409, respectively) also nullified enzymatic activity. On the basis of their critical roles in enzymatic activity and their sequence conservation, we propose that these motifs mediate sterol and acyl-CoA binding by this class of enzymes. PMID- 11483631 TI - Amyloid beta-peptide1-40 increases neuronal membrane fluidity: role of cholesterol and brain region. AB - There is increasing evidence of an interaction between cholesterol dynamics and Alzheimer's disease (AD), and amyloid beta-peptide may play an important role in this interaction. Abeta destabilizes brain membranes and this action of Abeta may be dependent on the amount of membrane cholesterol. We tested this hypothesis by examining effects of Abeta1-40 on the annular fluidity (i.e., lipid environment adjacent to proteins) and bulk fluidity of rat synaptic plasma membranes (SPM) of the cerebral cortex, cerebellum, and hippocampus using the fluorescent probe pyrene and energy transfer. Amounts of cholesterol and phospholipid of SPM from each brain region were determined. SPM of the cerebellum were significantly more fluid as compared with SPM of the cerebral cortex and hippocampus. Abeta significantly increased (P < or = 0.01) annular and bulk fluidity in SPM of cerebral cortex and hippocampus. In contrast, Abeta had no effect on annular fluidity and bulk fluidity of SPM of cerebellum. The amounts of cholesterol in SPM of cerebral cortex and hippocampus were significantly higher (P < or = 0.05) than amount of cholesterol in SPM of cerebellum. There was significantly less (P < or = 0.05) total phospholipid in cerebellar SPM as compared with SPM of cerebral cortex. Neuronal membranes enriched in cholesterol may promote accumulation of Abeta by hydrophobic interaction, and such an interpretation is consistent with recent studies showing that soluble Abeta can act as a seed for fibrillogenesis in the presence of cholesterol. PMID- 11483632 TI - High prevalence of low HDL cholesterol concentrations and mixed hyperlipidemia in a Mexican nationwide survey. AB - The prevalence of lipid abnormalities revealed in a survey done in 417 Mexican cities is described. Information was obtained on 15,607 subjects, aged 20 to 69 years. In this report, only samples obtained after a 9- to 12-h fast were included (2,256 cases: 953 men and 1,303 women). The population is representative of Mexican urban adults. Mean lipid concentrations were: cholesterol, 4.80 mmol/l; triglycerides, 2.39 mmol/l; HDL cholesterol, 1.00 mmol/l; and LDL cholesterol, 3.06 mmol/l. The most prevalent abnormality was HDL cholesterol below 0.9 mmol/l (46.2% for men and 28.7% for women). Hypertriglyceridemia (>2.26 mmol/l) was the second most prevalent abnormality (24.3%). Severe hypertriglyceridemia (>11.2 mmol/l) was observed in 0.42% of the population. Increased LDL cholesterol (> or =4.21 mmol/l) was observed in 11.2% of the sample. Half of the hypertriglyceridemic subjects had a mixed dyslipidemia or low HDL cholesterol. More than 50% of the low HDL cholesterol cases were not related to hypertriglyceridemia. Insulin resistance was found in 59% of them. In conclusion, the prevalence of hypoalphalipoproteinemia and other forms of dyslipidemia in Mexican adults is very high and it is among the highest previously reported worldwide. PMID- 11483633 TI - A cell-free assay for detecting HDL that is dysfunctional in preventing the formation of or inactivating oxidized phospholipids. AB - We have developed a novel and rapid cell-free assay of the ability of HDL to prevent the formation of or inactivate oxidized phospholipids. HDL was tested for its ability to inhibit the oxidation of LDL, or inhibit the oxidation of l-alpha 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (PAPC) by hydroperoxyoctadecadienoic acid (HPODE), or inactivate oxidized PAPC (Ox-PAPC). In each case the fluorescent signal generated in the presence of the test substances and the test HDL was determined. As little as 2.5 microg of normal human HDL cholesterol significantly inhibited the fluorescent signal generated by Ox-PAPC; results did not differ regardless of whether the HDL was prepared by gel electrophoresis, fast protein liquid chromatography, or dextran sulfate precipitation. HDL from each of 27 patients with coronary atherosclerosis failed to inhibit the fluorescent signal generated by a control LDL, whereas HDL from each of 31 matched normal subjects with the same levels of HDL cholesterol significantly inhibited the signal. Results from an established cell-based assay (Navab, M., S. Hama, J. Cooke, G. M. Anantharamaiah, M. Chaddha, L. Jin, G. Subbanagounder, K. F. Faull, S. T. Reddy, N. E. Miller, and A. M. Fogelman. 2000. J. Lipid Res. 41: 1481-1494) were identical. HDL from the patients also failed to inhibit the fluorescent signal generated from PAPC plus HPODE (10 of 10 patients) whereas HDL from matched controls (8 of 8 patients) significantly inhibited the fluorescent signal. We conclude that this new assay has the potential to allow widespread testing of the hypothesis that HDL that is dysfunctional in preventing the formation or inactivating oxidized phospholipids may play an important role in the development of atherosclerosis. PMID- 11483634 TI - Preparation of deacetyl-, lyso-, and deacetyl-lyso-GM(3) by selective alkaline hydrolysis of GM3 ganglioside. AB - Three methods (using GM3 quantities ranging from a few milligrams to grams) have been developed to prepare, in high yield, the three derivatives of ganglioside GM3 [alpha-Neu5Ac-(2-3)-beta-Gal-(1-4)-beta-Glc-(1-1)-ceramide]: deacetyl-GM3 [alpha-Neu-(2-3)-beta-Gal-(1-4)-beta-Glc-(1-1)-ceramide], lyso-GM3 [alpha-Neu5Ac (2-3)-beta-Gal-(1-4)-beta-Glc-(1-1)-sphingosine], and deacetyl-lyso-GM3 [alpha Neu-(2-3)-beta-Gal-(1-4)-beta-Glc-(1-1)-sphingosine]. This is the first report of the preparation of lyso-GM3 by a one-pot reaction. We can now define the optimal conditions for the different preparations. Preparation of deacetyl-GM3: alkaline reagent, 2 M KOH in water; GM3 concentration, 33 mg/ml; reaction temperature, 90 degrees C; reaction time, 3.5 h; nitrogen atmosphere. Preparation of deacetyl lyso-GM3: alkaline reagent, 8 M KOH in water; GM3 concentration, 10 mg/ml; reaction temperature, 90 degrees C; reaction time, 18 h; nitrogen atmosphere. Preparation of lyso-GM(3): alkaline reagent, 1 M sodium tert-butoxide in methanol; GM3 concentration, 10 mg/ml; reaction temperature, 80 degrees C; reaction time, 18 h; anhydrous conditions. The percentage yield of deacetyl-GM3 was 70;-75%, that of deacetyl-lyso-GM3 100%, and of lyso-GM3 36;-40%.Deacetyl GM3, deacetyl-lyso-GM3, and lyso-GM3 were purified by column chromatography, and chemical structures were confirmed by electron spray-mass spectrometry. PMID- 11483635 TI - Long-chain fatty acids modify hypertrophic responses of cultured primary neonatal cardiomyocytes. AB - In vivo, the normal heart obtains at least 60% of its energy from lipids and the remainder from glucose. Several lines of evidence indicate that an increase in the utilization of glucose [at the expense of fatty acids (FA)] may play a role in the genesis of hypertrophy. Primary cultures of neonatal cardiomyocytes have been used extensively to study the phenotype of these cells as well as their responses to hormonal hypertrophic agents. Unfortunately, such cultures are most typically cultured in glucose-rich FA-free media, and thus might be hypertrophied to start with. We therefore tested the effects of FA-albumin complexes on three different surrogate end points of hypertrophy of cardiomyocytes. Oleate-albumin complexes decreased the baseline values of all three variables, and increased the relative response of these variables to administration of norepinephrine. Oleate:palmitate-albumin complexes also affected all three variables and their responses to norepinephrine, but the effects differed somewhat from that of oleate-albumin complexes. Our results suggest that addition of long-chain FA, by providing conditions that more closely resemble physiological situations, may optimize the expression of hypertrophic responses in such cells. However, the differences between the effects of oleate and oleate:palmitate also suggest that the precise composition of FA may affect the phenotype of cardiomyocytes and how these cells respond to hypertrophic agents. PMID- 11483636 TI - A new method for HDL particle sizing by polyacrylamide gradient gel electrophoresis using whole plasma. AB - Low plasma levels of HDL cholesterol have been associated with an increased risk of coronary heart disease. HDL particles are heterogeneous with respect to size and apolipoprotein content. The objective of the present study was to develop a method to generate lipid-stainable calibrators that would allow the assessment of HDL particle size from whole plasma, using polyacrylamide gradient gel electrophoresis (PAGGE). Lipid-stainable HDL calibrators were obtained by subjecting isolated red blood cells to hemolysis either by freezing at -20 or -80 degrees C overnight or by rapid exposure to liquid nitrogen and mixing of the hemolysis products with plasma aliquots. All three methods were highly reproducible in producing Sudan black lipid-stainable HDL calibrators ranging from 75 to 200 A. The assessment of HDL particle size with these lipid-stainable HDL calibrators was also highly reproducible, with a coefficient of variation below 5.5%. These lipid-stainable HDL calibrators simplify the assessment of HDL particle size by PAGGE using whole plasma, without the need for costly, time consuming ultracentrifugation procedures. PMID- 11483637 TI - Complex and segmental uniparental disomy (UPD): review and lessons from rare chromosomal complements. AB - OBJECTIVE: To review all cases with segmental and/or complex uniparental disomy (UPD), to study aetiology and mechanisms of formation, and to draw conclusions. DESIGN: Searching published reports in Medline. RESULTS: The survey found at least nine cases with segmental UPD and a normal karyotype, 22 cases with UPD of a whole chromosome and a simple or a non-homologous Robertsonian translocation, eight cases with UPD and two isochromosomes, one of the short arm and one of the long arm of a non-acrocentric chromosome, 39 cases with UPD and an isochromosome of the long arm of two homologous acrocentric chromosomes, one case of UPD and an isochromosome 8 associated with a homozygous del(8)(p23.3pter), and 21 cases with UPD of a whole or parts of a chromosome associated with a complex karyotype. Segmental UPD is formed by somatic recombination (isodisomy) or by trisomy rescue. In the latter mechanism, a meiosis I error is associated with meiotic recombination and an additional somatic exchange between two non-uniparental chromatids. Subsequently, the chromatid that originated from the disomic gamete is lost (iso- and heterodisomy). In cases of UPD associated with one isochromosome of the short arm and one isochromosome of the long arm of a non acrocentric chromosome and in cases of UPD associated with a true isochromosome of an acrocentric chromosome, mitotic complementation is assumed. This term describes the formation by misdivision at the centromere during an early mitosis of a monosomic zygote. In cases of UPD associated with an additional marker chromosome, either mitotic formation of the marker chromosome in a trisomic zygote or fertilisation of a gamete with a marker chromosome formed in meiosis by a disomic gamete or by a normal gamete and subsequent duplication are possible. CONCLUSIONS: Research in the field of segmental and/or complex UPD may help to explain undiagnosed non-Mendelian disorders, to recognise hotspots for meiotic and mitotic recombinations, and to show that chromosomal segregation is more complex than previously thought. It may also be helpful to map autosomal recessively inherited genes, genes/regions of genomic imprinting, and dysmorphic phenotypes. Last but not least it would improve genetic counselling. PMID- 11483638 TI - VHL c.505 T>C mutation confers a high age related penetrance but no increased overall mortality. AB - BACKGROUND: Germline mutations of the VHL gene cause von Hippel-Lindau syndrome (VHL). In southern Germany, a specific mutation in this gene, c.505 T>C, is one of the most frequent alterations owing to a founder effect. METHODS: This study was conducted to evaluate morbidity, specific clinical risk profile, and mortality among a series of VHL c.505 T/C mutation carriers. A total of 125 eligible subjects carrying VHL c.505 T/C underwent ophthalmoscopy and gadolinium enhanced magnetic resonance imaging of the brain, the spinal cord, and the abdomen. Age related penetrance, morbidity, and mortality were assessed. RESULTS: Frequently observed lesions were phaeochromocytoma (47%), retinal angiomas (36%), haemangioblastoma of the spine (36%), and haemangioblastoma of the brain (16%). Four patients developed renal cell carcinoma. VHL was symptomatic in 47% of subjects; 30% were asymptomatic despite the presence of at least one VHL related tumour and 23% of the carriers had no detectable VHL lesion. Of the 19 patients who had died (15%), 10 died of symptomatic VHL lesions. Overall penetrance by cumulative incidence functions is estimated at 48% by 35 years and 88% by 70 years. In contrast to the only existing published report based on patients with presumably unselected VHL germline mutations, the mortality rate for c.505 T/C mutation carriers is comparable to that of the general population of Germany. CONCLUSIONS: Our results are an important example that a specific genotype, at least in the case of VHL c.505 T/C, can favourably impact on mortality despite a high age related penetrance. Our study also indirectly provides objective data which might be useful to the life and health insurance industry; it would appear that c.505 T>C mutation positive subjects have similar disease specific mortality to that of the general population owing to a combination of phenotype and timely detection of mutation carrier status followed by aggressive clinical screening and, if necessary, treatment. PMID- 11483639 TI - A common founder for the 35delG GJB2 gene mutation in connexin 26 hearing impairment. AB - Fifty to eighty percent of autosomal recessive congenital severe to profound hearing impairment result from mutations in a single gene, GJB2, that encodes the protein connexin 26. One mutation of this gene, the 35delG allele, is particularly common in white populations. We report evidence that the high frequency of this allelic variant is the result of a founder effect rather than a mutational hot spot in GJB2, which was the prevailing hypothesis. Patients homozygous for the 35delG mutation and normal hearing controls originating from Belgium, the UK, and the USA were genotyped for different single nucleotide polymorphisms (SNPs). Four SNPs mapped in the immediate vicinity of GJB2, while two were positioned up to 76 kb from it. Significant differences between the genotypes of patients and controls for the five SNPs closest to GJB2 were found, with nearly complete association of one SNP allele with the 35delG mutation. For the most remote SNP, we could not detect any association. We conclude that the 35delG mutation is derived from a common, albeit ancient founder. PMID- 11483640 TI - Predictive genetic testing in children and adults: a study of emotional impact. AB - AIM: To determine whether, following predictive genetic testing for familial adenomatous polyposis (FAP), children or adults receiving positive results experience clinically significant levels of anxiety or depression, and whether children receiving positive results experience higher levels of anxiety or depression than adults receiving positive results. DESIGN: Two studies, one cross sectional and one prospective. SAMPLE: 208 unaffected subjects (148 adults and 60 children) at risk for FAP who have undergone genetic testing since 1990. MAIN MEASURES: DEPENDENT VARIABLES: anxiety, depression; independent variables: test results, demographic measures, psychological resources (optimism, self-esteem). RESULTS: Study 1. In children receiving positive results, mean scores for anxiety and depression were within the normal range. There was a trend for children receiving positive results to be more anxious and depressed than those receiving negative results. In adults, mean scores for anxiety were within the normal range for those receiving negative results, but were in the clinical range for those receiving positive results, with 43% (95% CI 23-65) of the latter having scores in this range. Regardless of test result, adults were more likely to be clinically anxious if they were low in optimism or self-esteem. Children receiving positive or negative results did not experience greater anxiety or depression than adults. Study 2. For children receiving a positive test result, mean scores for anxiety, depression, and self-esteem were unchanged over the year following the result, while mean anxiety scores decreased and self-esteem increased after receipt of a negative test result over the same period of time. CONCLUSION: Children, as a group, did not show clinically significant distress over the first year following predictive genetic testing. Adults were more likely to be clinically anxious if they received a positive result or were low in optimism or self-esteem, with interacting effects. The association between anxiety, self-esteem, and optimism suggests that counselling should be targeted, not only at those with positive test results, but also at those low in psychological resources. PMID- 11483641 TI - A novel mutation in a family with non-syndromic sensorineural hearing loss that disrupts the newly characterised OTOF long isoforms. PMID- 11483642 TI - MET mutation and familial gastric cancer. PMID- 11483643 TI - Agenesis of cruciate ligaments and menisci causing severe knee dysplasia in TAR syndrome. PMID- 11483644 TI - Changes in mACh, NMDA and GABA(A) receptor binding after lateral fluid-percussion injury: in vitro autoradiography of rat brain frozen sections. AB - Adult rats were subjected to a moderate lateral fluid percussion injury (FPI), followed by survival periods of 2 and 12 h. Regional NMDA subtype glutamate, muscarinic acetylcholine and GABA(A) receptor binding in various brain regions was analysed by quantitative in vitro autoradiography and short-lived positron emission tomography tracers [11C]cyano-dizocilpine, 4-N [11C]methylpiperidylbenzilate (4-N-[11C]MPB), and [11C]flumazenil, respectively. The binding potential (BP, Bmax/KD) was calculated. The data with [11C]cyano dizocilpine showed a significant decrease in BP bilaterally for the frontoparietal cortex and hippocampus at both time points, in comparison with that of the sham-operated controls. At 12 h the decrease was significantly more prominent for the ipsilateral cortex and hippocampus than for the contralateral side. The BP of 4-N-[11C]MPB was significantly decreased after 2 h for the trauma side hippocampus, and after 12 h it had decreased for the trauma-site cortex and the bilateral hippocampus. The [11C]flumazenil exhibited a significant decrease in BP for the trauma-site cortex and the underlying hippocampus by 2 h after the traumatic brain injury. After 12 h a significantly decreased BP was observed only for the trauma-site cortex. The finding of a decreased BP demonstrates the involvement of these receptor systems in the development of cellular dysfunction, which is widespread and not limited to the site of lateral FPI. PMID- 11483645 TI - GABA(A) receptor alpha-subunit proteins in human chronic alcoholics. AB - Antibodies were raised against specific peptides from N-terminal regions of the alpha1 and alpha3 isoforms of the GABA(A) receptor, and used to assess the relative expression of these proteins in the superior frontal and primary motor cortices of 10 control, nine uncomplicated alcoholic and six cirrhotic alcoholic cases were matched for age and post-mortem delay. The regression of expression on post-mortem delay was not statistically significant for either isoform in either region. In both cortical areas, the regression of alpha1 expression on age differed significantly between alcoholic cases, which showed a decrease, and normal controls, which did not. Age had no effect on alpha3 expression. The alpha1 and alpha3 isoforms were found to be expressed differentially across cortical regions and showed a tendency to be expressed differentially across case groups. In cirrhotic alcoholics, alpha1 expression was greater in superior frontal than in motor cortex, whereas this regional difference was not significant in controls or uncomplicated alcoholics. In uncomplicated alcoholics, alpha3 expression was significantly lower in superior frontal than in motor cortex. Expression of alpha1 was significantly different from that of alpha3 in the superior frontal cortex of alcoholics, but not in controls. In motor cortex, there were no significant differences in expression between the isoforms in any case group. PMID- 11483646 TI - Neuroprotection mediated by glial group-II metabotropic glutamate receptors requires the activation of the MAP kinase and the phosphatidylinositol-3-kinase pathways. AB - The mGlu2/3 receptor agonists 4-carboxy-3-hydroxyphenylglycine (4C3HPG) and LY379268 attenuated NMDA toxicity in primary cultures containing both neurons and astrocytes. Neuroprotection was abrogated by PD98059 and LY294002, which inhibit the mitogen activated protein kinase (MAPK) and the phosphatidylinositol-3-kinase (PI-3-K) pathways, respectively. Cultured astrocytes lost the ability to produce transforming growth factor-beta1 (TGF-beta1) in response to mGlu2/3 receptor agonists when co-incubated with PD98059 or LY294002. As a result, the glial medium was no longer protective against NMDA toxicity. Activation of the MAPK and PI-3-K pathways in cultured astrocytes treated with 4C3HPG or LY379268 was directly demonstrated by an increase in the phosphorylated forms of ERK-1/2 and Akt. Similarly to that observed in the culture, intracerebral or systemic injections of mGlu2/3 receptor agonists enhanced TGF-beta1 formation in the rat or mouse caudate nucleus, and this effect was reduced by PD98059. PD98059 also reduced the ability of LY379268 to protect striatal neurons against NMDA toxicity. These results suggest that activation of glial mGlu2/3 receptors induces neuroprotection through the activation of the MAPK and PI-3-K pathways leading to the induction of TGF-beta. PMID- 11483647 TI - Identification and functional characterization of thromboxane A2 receptors in Schwann cells. AB - Previous reports have demonstrated the presence of functional thromboxane A2 (TP) receptors in astrocytes and oligodendrocytes. In these experiments, the presence and function of TP receptors in primary rat Schwann cells (rSC) and a neurofibrosarcoma-derived human Schwann cell line (T265) was investigated. Immunocytochemical and immunoblot analyses using polyclonal anti-TP receptor antibodies demonstrate that both cell types express TP receptors. Treatment with the stable thromboxane A2 mimetic U46619 (10 microM) did not stimulate intracellular calcium mobilization in rSC, whereas T265 cells demonstrated a calcium response that was inhibited by prior treatment with TP receptor antagonists. U46619 also stimulated CREB phosphorylation on Ser133 in T265 cells and, to a lesser extent, in rSC. To identify potential mechanisms of CREB phosphorylation in rSC, we monitored intracellular cAMP levels following U46619 stimulation. Elevated levels of cAMP were detected in both rSC (20-fold) and T265 (15-fold) cells. These results demonstrate that TP receptor activation specifically stimulates CREB phosphorylation in T265 cells, possibly by a calcium and/or cAMP-dependent mechanism. In contrast, TP receptor activation in rSC stimulates increases in cAMP and CREB phosphorylation but does not elicit changes in intracellular calcium. PMID- 11483648 TI - HIV-1 Tat through phosphorylation of NMDA receptors potentiates glutamate excitotoxicity. AB - Toxic effects of HIV-1 proteins contribute to altered function and decreased survival of select populations of neurons in HIV-1-infected brain. One such HIV-1 protein, Tat, can activate calcium release from IP3-sensitive intracellular pools, induce calcium influx in neural cells, and, as a result, can increase neuronal cell death. Here, we provide evidence that Tat potentiates excitatory amino acid (glutamate and NMDA) triggered calcium flux, as well as glutamate- and staurosporine-mediated neurotoxicity. Calcium flux in cultured rat hippocampal neurons triggered by the transient application of glutamate or NMDA was facilitated by pre-exposure to Tat. Facilitation of glutamate-triggered calcium flux by Tat was prevented by inhibitors of ADP-ribosylation of G(i)/G(o) proteins (pertussis toxin), protein kinase C (H7 and bisindolymide), and IP3-mediated calcium release (xestospongin C), but was not prevented by an activator of G(s) (cholera toxin) or an inhibitor of protein kinase A (H89). Facilitation of NMDA triggered calcium flux by Tat was reversed by inhibitors of tyrosine kinase (genestein and herbimycin A) and by an inhibitor of NMDA receptor function (zinc). Tat increased 32P incorporation into NMDA receptor subunits NR2A and NR2B and this effect was blocked by genestein. Subtoxic concentrations of Tat combined with subtoxic concentrations of glutamate or staurosporine increased neuronal cell death significantly. Together, these findings suggest that NMDA receptors play an important role in Tat neurotoxicity and the mechanisms identified may provide additional therapeutic targets for the treatment of HIV-1 associated dementia. PMID- 11483649 TI - The inhibition of phosphatidylinositol-3-kinase induces neurite retraction and activates GSK3. AB - It has been extensively described that neuronal differentiation involves the signalling through neurotrophin receptors to a Ras-dependent mitogen-activated protein kinase (MAPK) cascade. However, signalling pathways from other neuritogenic factors have not been well established. It has been reported that cAMP may activate protein kinase (PKA), and it has been shown that PKA-mediated stimulation of MAPK pathway regulates not only neuritogenesis but also survival. However, extracellular regulated kinases (ERKs) mediated pathways are not sufficient to explain all the processes which occur in neuronal differentiation. Our present data show that: in cAMP-mediated neuritogenesis, using the SH-SY5Y human neuroblastoma cell line, there exists a link between the activation of PKA and stimulation of phosphatidylinositol 3-kinase (PI3K). Both kinase activities are essential to the initial elongation steps. Surprisingly, this neuritogenic process appears to be independent of ERKs. While the activity of PI3K is essential for elongation and maintenance of neurites, its inhibition causes retraction. In this neurite retraction process, GSK3 is activated. Using both a pharmacological approach and gene transfer of a dominant negative form of GSK3, we conclude that this induced retraction is a GSK3-dependent process which in turn appears to be a common target for transduction pathways involved in lysophosphatidic acid-mediated and PI3K-mediated neurite retraction. PMID- 11483650 TI - Semaphorin4F interacts with the synapse-associated protein SAP90/PSD-95. AB - Semaphorins are a family of secreted and membrane-associated proteins involved in growth cone guidance during development. Here, we describe the interaction of Semaphorin4F (Sema4F) with the post-synaptic density protein SAP90/PSD-95. Using the yeast two-hybrid system and coprecipitation assays we were able to show an interaction between the extreme C-terminus of Sema4F and the PDZ domains of SAP90/PSD-95. Heterologous coexpression of a chimeric EphrinB1/Semaphorin4F protein with SAP90/PSD-95 in COS cells leads to translocation of SAP90/PSD-95 from the cytosol to the membrane. Deletion analysis shows that this translocation activity of Sema4F is completely dependent on the presence of the last three C terminal amino acids. In addition, Sema4F immunoreactivity is present in synaptosome fractions and enriched in post-synaptic density fractions. Consistently, in cultured hippocampal neurons, we demonstrate punctate colocalization of Sema4F and SAP90/PSD-95 in dendrites, furthermore we found colocalization of Sema4F with synapsin1 suggesting a synaptic localization. Our data implicate a new functional context for semaphorins at glutamatergic synapses. PMID- 11483651 TI - Tyrosine hydroxylase phosphorylation in bovine adrenal chromaffin cells: the role of MAPKs after angiotensin II stimulation. AB - Angiotensin II (AII, 100 nM) stimulation of bovine adrenal chromaffin cells (BACCs) produced angiotensin II receptor subtype 1 (AT1)-mediated increases in extracellular regulated protein kinase 1/2 (ERK1/2) and stress-activated p38MAPK (p38 kinase) phosphorylation over a period of 10 min. ERK1/2 and p38 kinase phosphorylation preceded Ser31 phosphorylation on tyrosine hydroxylase (TOH). The inhibitors of mitogen-activated protein kinase kinase 1/2 (MEK1/2) activation, PD98059 (0.1-50 microM) and UO126 (0.1-10 microM), dose-dependently inhibited both ERK2 and Ser31 phosphorylation on TOH in response to AII, suggesting MEK1/2 involvement. The p38 kinase inhibitor SB203580 (20 microM, 30 min) abolished Ser31 and Ser19 phosphorylation on TOH and partially inhibited ERK2 phosphorylation produced by AII. In contrast, 1 microM SB203580 did not affect AII-stimulated TOH phosphorylation, but fully inhibited heat shock protein 27 (HSP27) phosphorylation produced by AII. Also, 1 microM SB203580 fully inhibited Ser19 phosphorylation on TOH and HSP27 phosphorylation in response to anisomycin (30 min, 10 microg/mL). The results suggest that ERKs mediate Ser31 phosphorylation on TOH in response to AII, but p38 kinase is not involved. Previous studies suggesting a role for p38 kinase in the phosphorylation of Ser31 are explained by the non-specific effects of 20 microM SB203580 in BACCs. The p38 kinase pathway is able to phosphorylate Ser19 on TOH in response to anisomycin, but does not do so in response to AII. PMID- 11483652 TI - Calcium and pituitary adenylate cyclase-activating polypeptide induced expression of circadian clock gene mPer1 in the mouse cerebellar granule cell culture. AB - Mammalian circadian clock genes Per1 and Per2 are rhythmically expressed not only in the suprachiasmatic nucleus where the mammalian circadian clock exists, but also in other brain regions and peripheral tissues. The induced circadian oscillation of Per genes after treatment with high concentrations of serum or various drugs in cultured cells suggests the ubiquitous existence of the oscillatory mechanism. These treatments also result in a rapid surge of expression of Per1. It has been shown that multiple signaling pathways are involved in Per1 gene induction in culture cells. We used a dispersed primary cell culture made up of mouse cerebellar granule cells to examine the stimuli inducing the mPer genes and their signaling pathways in neuronal tissues expressing mPer genes. We demonstrated that mPer1, but not mPer2, mRNA expression was dependent on the depolarization state controlled by extracellular KCl concentration in the granule cell culture. Nifedipine treatment reduced mPer1 induction, suggesting that mPer1 mRNA expression depends on intracellular calcium concentration regulated through a voltage-dependent Ca2+ channel. Transient mPer1 mRNA induction was observed after elevating KCl concentration in the medium from 5 mM to 25 mM. This increased expression was suppressed by a calmodulin antagonist, or CaMKII/IV inhibitor, but not by MEK inhibitors. Addition of pituitary adenylate cyclase-activating polypeptide-38 to the medium also induced transient Per1 gene expression. This induction was mimicked by dibutyryl-cAMP and suppressed by a protein kinase A (PKA) inhibitor, but not by MEK inhibitors. These results suggest that Ca2+/calmodulin-dependent protein kinase II/IV- and PKA-dependent pathways are involved in high-KCl and PACAP-induced mPer1 induction, respectively, and neural tissues use multiple signaling pathways for mPer1 induction similar to culture cells. PMID- 11483653 TI - 6-hydroxydopamine increases the hydroxylation and nitration of phenylalanine in vivo: implication of peroxynitrite formation. AB - In the present study, we investigated the effect of the dopaminergic neurotoxin 6 hydroxydopamine (6-OHDA) on hydroxyl free radical and peroxynitrite formation in vivo using D-phenylalanine as a novel mechanistic probe. In vivo microdialysis was carried out in the striatum of freely moving male Wistar rats. The microdialysis probes were perfused with artificial cerebrospinal fluid containing 5 mM D-phenylalanine (flow rate 2 microL/min). After obtaining a stable baseline 6-OHDA was delivered into the striatum via reverse microdialysis for 60 min. HPLC measurements of the effluent were performed using photodiode array detection for determination of phenylalanine derived o-tyrosine and m-tyrosine (as hydroxylation markers) as well as of nitrotyrosine and nitrophenylalanine (as nitration markers). The basal levels of the hydroxylation derived products of phenylalanine were approximately 100-fold higher than those of the nitration derived products. 6-OHDA (0.1, 1, 10 mM) significantly increased o- and m tyrosine up to nine- and 13-fold, respectively, whereas levels of 3-nitrotyrosine and 4-nitrophenylalanine were significantly increased up to 422- and 358-fold, respectively. The results demonstrate that phenylalanine is a sensitive in vivo marker for 6-OHDA-induced hydroxylation and nitration reactions which are clearly concentration dependent. We conclude that peroxynitrite formation is involved in 6-OHDA-induced neurochemical effects. PMID- 11483654 TI - The binding of zinc and copper ions to nerve growth factor is differentially affected by pH: implications for cerebral acidosis. AB - It has recently been shown that transition metal cations Zn2+ and Cu2+ bind to histidine residues of nerve growth factor (NGF) and other neurotrophins (a family of proteins important for neuronal survival) leading to their inactivation. Experimental data and theoretical considerations indicate that transition metal cations may destabilize the ionic form of histidine residues within proteins, thereby decreasing their pK(a) values. Because the release of transition metal cations and acidification of the local environment represent important events associated with brain injury, the ability of Zn2+ and Cu2+ to bind to neurotrophins in acidic conditions may alter neuronal death following stroke or as a result of traumatic injury. To test the hypothesis that metal ion binding to neurotrophins is influenced by pH, the effects of Zn2+ and Cu2+ on NGF conformation, receptor binding and NGF tyrosine kinase (trkA) receptor signal transduction were examined under conditions mimicking cerebral acidosis (pH range 5.5-7.4). The inhibitory effect of Zn2+ on biological activities of NGF is lost under acidic conditions. Conversely, the binding of Cu2+ to NGF is relatively independent of pH changes within the studied range. These data demonstrate that Cu2+ has greater binding affinity to NGF than Zn2+ at reduced pH, consistent with the higher affinity of Cu2+ for histidine residues. These findings suggest that cerebral acidosis associated with stroke or traumatic brain injury could neutralize the Zn2+-mediated inactivation of NGF, whereas corresponding pH changes would have little or no influence on the inhibitory effects of Cu2+. The importance of His84 of NGF for transition metal cation binding is demonstrated, confirming the involvement of this residue in metal ion coordination. PMID- 11483655 TI - Tyrosine phosphorylation of the N-methyl-D-aspartate receptor by exogenous and postsynaptic density-associated Src-family kinases. AB - Phosphorylation of the NMDA receptor by Src-family tyrosine kinases has been implicated in the regulation of receptor function. We have investigated the tyrosine phosphorylation of NMDA receptor subunits NR2A and NR2B by exogenous Src and Fyn and compared this to phosphorylation by tyrosine kinases associated with the postsynaptic density (PSD). Phosphorylation of the receptor by exogenous Src and Fyn was dependent upon initial binding of the kinases to PSDs via their SH2 domains. Src and Fyn phosphorylated similar sites in NR2A and NR2B, tryptic peptide mapping identifying seven and five major tyrosine-phosphorylated peptides derived from NR2A and NR2B, respectively. All five tyrosine phosphorylation sites on NR2B were localized to the C-terminal, cytoplasmic domain. Phosphorylation of NR2B by endogenous PSD tyrosine kinases yielded only three tyrosine phosphorylated tryptic peptides, two of which corresponded to Src phosphorylation sites, and one of which was novel. Phosphorylation-site specific antibodies identified NR2B Tyr1472 as a phosphorylation site for intrinsic PSD tyrosine kinases. Phosphorylation of this site was inhibited by the Src-family-specific inhibitor PP2. The results identify several potential phosphorylation sites for Src in the NMDA receptor, and indicate that not all of these sites are available for phosphorylation by kinases located within the structural framework of the PSD. PMID- 11483656 TI - L-DOPA and glia-conditioned medium have additive effects on tyrosine hydroxylase expression in human catecholamine-rich neuroblastoma NB69 cells. AB - The aim of this study was to investigate the effect of L-DOPA and glia conditioned medium (GCM) on cell viability, tyrosine hydroxylase (TH) expression, dopamine (DA) metabolism and glutathione (GSH) levels of NB69 cells. L-DOPA (200 microM) induced differentiation of NB69 cells of more than 4 weeks in vitro, as shown by phase-contrast microscopy and TH immunocytochemistry, and decreased replication, as shown by 5-bromodeoxyuridine immunostaining. L-DOPA did not increase the number of necrotic or apoptotic cells, as shown by morphological features, Trypan Blue, lactate dehydrogenase activity, bis-benzimide staining and TUNEL assay. Furthermore, L-DOPA (200 microM) increased Bcl-xL protein expression. Incubation of cells with L-DOPA (50, 100, 200 microM) for 24 h resulted in an increase in TH protein levels (174, 196 and 212% versus control). Neither carbidopa, an inhibitor of L-aromatic amino acid decarboxylase enzyme, nor L-buthionine sulfoximine, which inhibits GSH synthesis, or ascorbic acid, an antioxidant, blocked the L-DOPA-induced effect on TH protein expression. L-DOPA (0, 50, 100 and 200 microM) plus GCM further increased the amount of TH protein (346, 446, 472 and 424%). L-DOPA (200 microM) increased TH protein levels to 132, 191 and 245% of controls after incubation for 24, 48 and 72 h. DA metabolism in NB69 cells was increased in cultures treated with either L-DOPA (200-300 microM) or GCM and these two agents had a synergistic effect on DA metabolism. In addition, L-DOPA (200 microM) or/and GCM-treated cells increased their GSH extracellular levels (223, 257, 300% of controls) after 48 h of treatment. The L DOPA-induced increase of TH protein expression in NB69 cells was independent of DA production, free radicals and GSH up-regulation. PMID- 11483657 TI - Agonist-induced internalization of the metabotropic glutamate receptor 1a is arrestin- and dynamin-dependent. AB - At present, little is known regarding the mechanism of metabotropic glutamate receptor (mGluR) trafficking. To facilitate this characterization we inserted a haemagglutinin (HA) epitope tag in the extracellular N-terminal domain of the rat mGluR1a. In human embryonic kidney cells (HEK293), transiently transfected with HA-mGluR1a, the epitope-tagged receptor was primarily localized to the cell surface prior to agonist stimulation. Following stimulation with glutamate (10 microM; 30 min) the HA-mGluR1a underwent internalization to endosomes. Further quantification of receptor internalization was provided by ELISA experiments which showed rapid agonist-induced internalization of the HA-mGluR1a. To determine whether agonist-induced mGluR1a internalization is an arrestin- and dynamin-dependent process, cells were cotransfected with HA-mGluR1a and either of these dynamin-K44A or arrestin-2 (319-418). Expression of either dominant negative mutant constructs with receptor strongly inhibited glutamate-induced (10 microM; 30 min) HA-mGluR1a internalization. In addition, wild-type arrestin-2 green fluorescent protein (arrestin-2-GFP) or arrestin-3-GFP underwent agonist induced translocation from cytosol to membrane in HEK293 cells coexpressing HA mGluR1a. Taken together our observations demonstrate that agonist-induced internalization of mGluR1a is an arrestin- and dynamin-dependent process. PMID- 11483658 TI - Regulation of functional nitric oxide synthase-1 expression in cerebellar granule neurons by heregulin is post-transcriptional, and involves mitogen-activated protein kinase. AB - The neuregulins, and the ErbB family of tyrosine kinase receptors, play important roles in the development of the nervous system. Recently, the C-terminal region of the ErbB4 receptor has been reported to associate with domains of post synaptic density proteins. The latter are, in turn, known to assemble nitric oxide synthase (NOS)-1 at cell junctions. Previously, we showed that heregulin can up-regulate the expression of NOS-1 via the ErbB4 receptor in cerebellar granule cell cultures. We have now determined that this up-regulation is post transcriptional, and results in an associated increase in NOS activity in these neurons. Furthermore, we find that heregulin activates both MAP kinase and phosphoinositide 3 kinase (PI 3-K) in granule cells. While inhibition of MAP kinase reduces the ability of heregulin to up-regulate NOS-1 expression, a specific inhibitor of PI 3-K was without effect. Our results suggest that NO could mediate some of the downstream effects of heregulin in the nervous system. PMID- 11483659 TI - Selective down-regulation of the astrocyte glutamate transporters GLT-1 and GLAST within the medial thalamus in experimental Wernicke's encephalopathy. AB - Although earlier studies on thiamine deficiency have reported increases in extracellular glutamate concentration in the thalamus, a vulnerable region of the brain in this disorder, the mechanism by which this occurs has remained unresolved. Treatment with pyrithiamine, a central thiamine antagonist, resulted in a 71 and 55% decrease in protein levels of the astrocyte glutamate transporters GLT-1 and GLAST, respectively, by immunoblotting in the medial thalamus of day 14 symptomatic rats at loss of righting reflexes. These changes occurred prior to the onset of convulsions and pannecrosis. Loss of both GLT-1 and GLAST transporter sites was also confirmed in this region of the thalamus at the symptomatic stage using immunohistochemical methods. In contrast, no change in either transporter protein was detected in the non-vulnerable frontal parietal cortex. These effects are selective; protein levels of the astrocyte GABA transporter GAT-3 were unaffected in the medial thalamus. In addition, astrocyte specific glial fibrillary acidic protein (GFAP) content was unchanged in this brain region, suggesting that astrocytes are spared in this disorder. Loss of GLT 1 or GLAST protein was not observed on day 12 of treatment, indicating that down regulation of these transporters occurs within 48 h prior to loss of righting reflexes. Finally, GLT-1 content was positively correlated with levels of the neurofilament protein alpha-internexin, suggesting that early neuronal drop-out may contribute to the down-regulation of this glutamate transporter and subsequent pannecrosis. A selective, focal loss of GLT-1 and GLAST transporter proteins provides a rational explanation for the increase in interstitial glutamate levels, and may play a major role in the selective vulnerability of thalamic structures to thiamine deficiency-induced cell death. PMID- 11483660 TI - Amyloid beta proteins inhibit Cl(-)-ATPase activity in cultured rat hippocampal neurons. AB - Cl(-)-ATPase in the CNS is a candidate for an outwardly directed neuronal Cl(-) transporter requiring phosphatidylinositol-4-phosphate (PI4P) for its optimal activity. To test its pathophysiological changes in a phosphatidylinositol (PI) metabolism disorder, the effects of neurotoxic factors in Alzheimer's disease (AD), amyloid beta proteins (Abetas), on the Cl(-)-ATPase activity were examined using primary cultured rat hippocampal neurons. Amyloid beta proteins (1-40, 1-42 and 25-35) concentration-dependently (1-100 nM) and time-dependently (from 1 h to 6 day) decreased Cl(-)-ATPase activity and elevated intracellular Cl(-) concentrations ([Cl(-)]i), Abeta25-35 being the most potent. Addition of inositol or 8-Br-cyclic GMP completely reversed these Abeta-induced changes. The recoveries in enzyme activity were attenuated by an inhibitor of PI 4-kinase, 10 microM wortmannin or 20 microM quercetin, but not by a PI 3-kinase inhibitor, 50 nM wortmannin or 10 microM LY294002. The PI, PIP and PIP2 levels of the plasma membrane-rich fraction were lower in the Abeta-treated cells as compared with each control. In the Abeta-exposed culture, but not in control, stimulation by 10 microM glutamate for 10 min significantly increased fragmentation of DNA and decreased cell viability. Addition of inositol or 8-Br-cyclic GMP prevented the effect of Abeta-treatment on the neurotoxicity of glutamate. Thus, Abetas reduce neuronal Cl(-)-ATPase activity, resulting in an increase in [Cl(-)]i probably by lowering PI4P levels, and this may reflect a pre-apoptotic condition in early pathophysiological profiles of AD. PMID- 11483661 TI - Selective and biphasic effect of the membrane lipid peroxidation product 4 hydroxy-2,3-nonenal on N-methyl-D-aspartate channels. AB - Increased oxyradical production and membrane lipid peroxidation occur in neurons under physiological conditions and in neurodegenerative disorders. Lipid peroxidation can alter synaptic plasticity and may increase the vulnerability of neurons to excitotoxicity, but the underlying mechanisms are unknown. We report that 4-hydroxy-2,3-nonenal (4HN), an aldehyde product of lipid peroxidation, exerts a biphasic effect on NMDA-induced current in cultured rat hippocampal neurons with current being increased during the first 2 h and decreased after 6 h. Similarly, 4HN causes an early increase and a delayed decrease in NMDA-induced elevation of intracellular Ca2+ levels. In contrast, 4HN affects neither the ion current nor the Ca2+ response to alpha-amino-3-hydroxy-5-methylisoxazole-4 propionate (AMPA). The initial enhancement of NMDA-induced current is associated with increased phosphorylation of the NR1 receptor subunit, whereas the delayed suppression of current is associated with cellular ATP depletion and mitochondrial membrane depolarization. Cell death induced by 4HN is attenuated by an NMDA receptor antagonist, but not by an AMPA receptor antagonist. A secreted form of amyloid precursor protein, previously shown to protect neurons against oxidative and excitotoxic insults, prevented each of the effects of 4HN including the early and late changes in NMDA current, delayed ATP depletion, and cell death. These findings show that the membrane lipid peroxidation product 4HN can modulate NMDA channel activity, suggesting a role for this aldehyde in physiological and pathophysiological responses of neurons to oxidative stress. PMID- 11483662 TI - Inorganic lead stimulates DNA synthesis in human astrocytoma cells: role of protein kinase Calpha. AB - As lead has been shown to activate protein kinase C (PKC), and gliomas are reported to be highly dependent on PKC for their proliferation, this study was undertaken to investigate whether lead may act as a mitogen in human astrocytoma cells, and to determine the role of PKC in this effect. Lead acetate (from 100 nM to 100 microM) induced a concentration- and time-dependent increase in DNA synthesis, as measured by incorporation of [methyl-3H]thymidine into cell DNA, without causing any cytotoxicity. Flow cytometric analysis showed that lead was able to stimulate the cell cycle transition from the G0/G1 phase to the S/G2 phase, resulting in increased percentage of cells in the latter phase. Western blot analyses showed that lead induced translocation of PKCalpha, but not of PKCepsilon or PKCzeta, from the cytosolic to the particulate fraction, with a concomitant increase in PKC enzyme activity. Prolonged exposure to lead caused down-regulation of PKCalpha, but not of PKCepsilon. The effect of lead on DNA synthesis was mediated through PKC as evidenced by the finding that two PKC inhibitors, GF 109203X and staurosporine, as well as down-regulation of PKC through prolonged treatment with 12-O-tetradecanoylphorbol 13-acetate, blocked lead-induced DNA synthesis. Further experiments using a pseudosubstrate peptide targeting classical PKCs and selective down-regulation of specific PKC isoforms indicated that the effect of lead on DNA synthesis was mediated by PKCalpha. Altogether, these results suggest that lead stimulates DNA synthesis in human astrocytoma cells by a mechanism that involves activation of PKCalpha. PMID- 11483663 TI - Zn2+-induced ERK activation mediated by reactive oxygen species causes cell death in differentiated PC12 cells. AB - Recent studies have provided evidence that Zn2+ plays a crucial role in ischemia- and seizure-induced neuronal death. However, the intracellular signaling pathways involved in Zn2+-induced cell death are largely unknown. In the present study, we investigated the roles of mitogen-activated protein kinases (MAPKs), such as c Jun N-terminal kinase (JNK), p38 MAPK and extracellular signal-regulated kinase (ERK), and of reactive oxygen species (ROS) in Zn2+-induced cell death using differentiated PC12 cells. Intracellular accumulation of Zn2+ induced by the combined application of pyrithione (5 microM), a Zn2+ ionophore, and Zn2+ (10 microM) caused cell death and activated JNK and ERK, but not p38 MAPK. Preventing JNK activation by the expression of dominant negative SEK1 (SEKAL) did not attenuate Zn2+-induced cell death, whereas the inhibition of ERK with PD98059 and the expression of dominant negative Ras mutant (RasN17) significantly prevented cell death. Inhibition of protein kinase C (PKC) and phosphatidylinositol-3 kinase had little effect on Zn2+-induced ERK activation. Intracellular Zn2+ accumulation resulted in the generation of ROS, and antioxidants prevented both the ERK activation and the cell death induced by Zn2+. Therefore, we conclude that although Zn2+ activates JNK and ERK, only ERK contributes to Zn2+-induced cell death, and that ERK activation is mediated by ROS via the Ras/Raf/MEK/ERK signaling pathway. PMID- 11483664 TI - Evidence that increases of mitochondrial immunoreactive IL-1beta by HIV-1 gp120 implicate in situ cleavage of pro-IL-1beta in the neocortex of rat. AB - Immunoelectron microscopy analysis of brain tissue sections and rat-specific sandwich ELISA allowed the localization of interleukin-1beta (IL-1beta) immunoreactivity in the mitochondria and cytosol of neocortical tissue preparations from the brain of naive, untreated, rats and rats receiving a single daily injection into one lateral cerebral ventricle (i.c.v.) of bovine serum albumin (BSA; 100 ng/day) for seven consecutive days. Interestingly, seven days i.c.v. treatment with the HIV-1 coat protein gp120 (100 ng/day) enhances IL-1beta immunoreactivity in the cellular fractions studied. Elevation of mitochondrial immunoreactive IL-1beta levels seems to originate from the conversion operated by the interleukin converting enzyme (ICE) of mitochondrial pro-IL-1beta; in fact, IL-1beta increases reported in the ELISA experiments were paralleled by a decrease of the mitochondrial pro-IL-1beta 31-kDa band in conjunction with enhanced expression of the p20 component of activated ICE. In conclusion, the present results demonstrate that gp120-enhanced neocortical expression of IL 1beta originates, at least in part, from in situ cleavage of mitochondrial pro-IL 1beta and suggest that this, together with the central role of the mitochondrion in the expression of programmed cell death, may be important for apoptosis induced by the viral coat protein in the brain of rats. PMID- 11483665 TI - Regional changes in density of serotonin transporter in the brain of 5-HT1A and 5 HT1B knockout mice, and of serotonin innervation in the 5-HT1B knockout. AB - 5-HT1A knockout (KO) mice display an anxious-like phenotype, whereas 5-HT1B KOs are over-aggressive. To identify serotoninergic correlates of these altered behaviors, autoradiographic measurements of 5-HT1A and 5-HT1B serotonin (5-HT) receptors and transporter (5-HTT) were obtained using the radioligands [3H]8-OH DPAT, [125I]cyanopindolol and [3H]citalopram, respectively. By comparison to wild type, density of 5-HT1B receptors was unchanged throughout brain in 5-HT1A KOs, and that of 5-HT1A receptors in 5-HT1B KOs. In contrast, decreases in density of 5-HTT binding were measured in several brain regions of both genotypes. Moreover, 5-HTT binding density was significantly increased in the amygdalo-hippocampal nucleus and ventral hippocampus of the 5-HT1B KOs. Measurements of 5-HT axon length and number of axon varicosities by quantitative 5-HT immunocytochemistry revealed proportional increases in the density of 5-HT innervation in these two regions of 5-HT1B KOs, whereas none of the decreases in 5-HTT binding sites were associated with any such changes. Several conclusions could be drawn from these results: (i) 5-HT1B receptors do not adapt in 5-HT1A KOs, nor do 5-HT1A receptors in 5-HT1B KOs. (ii) 5-HTT is down-regulated in several brain regions of 5-HT1A and 5-HT1B KO mice. (iii) This down-regulation could contribute to the anxious like phenotype of the 5-HT1A KOs, by reducing 5-HT clearance in several territories of 5-HT innervation. (iv) The 5-HT hyperinnervation in the amygdalo hippocampal nucleus and ventral hippocampus of 5-HT1B KOs could play a role in their increased aggressiveness, and might also explain their better performance in some cognitive tests. (v) These increases in density of 5-HT innervation provide the first evidence for a negative control of 5-HT neuron growth mediated by 5-HT1B receptors. PMID- 11483666 TI - The Ras-ERK pathway is required for the induction of neuronal nitric oxide synthase in differentiating PC12 cells. AB - We have studied the role of MAP kinase pathways in neuronal nitric oxide synthase (nNOS) induction during the differentiation of PC12 cells. In nerve growth factor (NGF)-treated PC12 cells, we find nNOS induced at RNA and protein levels, resulting in increased NOS activity. We note that neither nNOS mRNA, nNOS protein nor NOS activity is induced by NGF treatment in cells that have been infected with a dominant negative Ras adenovirus. We have also used drugs that block MAP kinase pathways and assessed their ability to inhibit nNOS induction. Even though U0126 and PD98059 are both MEK inhibitors, we find that U0126, but not PD98059, blocks induction of nNOS protein and NOS activity in NGF-treated PC12 cells. Also, the p38 kinase inhibitor, SB203580, does not block nNOS induction in our clone of PC12 cells. Since the JNK pathway is not activated in NGF-treated PC12 cells, we conclude that the Ras-ERK pathway and not the p38 or JNK pathway is required for nNOS induction in NGF-treated PC12 cells. We find that U0126 is much more effective than PD98059 in blocking the Ras-ERK pathway, thereby explaining the discrepancy in nNOS inhibition. We conclude that the Ras-ERK pathway is required for nNOS induction. PMID- 11483667 TI - Interleukin-1beta up-regulates expression of neurofilament light in human neuronal cells. AB - Elevated expression of interleukin-1 (IL-1beta), a pro-inflammatory cytokine secreted by activated microglia, is a pathogenic marker of numerous neurodegenerative processes including Alzheimer's disease (AD). We have characterized a link between IL-1beta and the 68-kDa neurofilament light (NF-L) protein, which is a major component of the neuronal cytoskeleton. Using human brain aggregate cultures, we found that IL-1beta treatment significantly increased NF-L expression in primary neurons. Analysis of mRNA levels demonstrated elevated NF-L expression within 72 h while imaging of neurons by immunofluorescent staining for NF-L confirmed IL-1beta-induced NF-L protein expression. These observations suggest a potential inflammatory-induced mechanism for deregulation of an important cytoskeletal protein, NF-L, possibly leading to neuronal dysfunction. PMID- 11483668 TI - Evidence of a role for cyclic ADP-ribose in calcium signalling and neurotransmitter release in cultured astrocytes. AB - Astrocytes possess different, efficient ways to generate complex changes in intracellular calcium concentrations, which allow them to communicate with each other and to interact with adjacent neuronal cells. Here we show that cultured hippocampal astrocytes coexpress the ectoenzyme CD38, directly involved in the metabolism of the calcium mobilizer cyclic ADP-ribose, and the NAD+ transporter connexin 43. We also demonstrate that hippocampal astrocytes can release NAD+ and respond to extracellular NAD+ or cyclic ADP-ribose with intracellular calcium increases, suggesting the existence of an autocrine cyclic ADP-ribose-mediated signalling. Cyclic ADP-ribose-induced calcium changes are in turn responsible for an increased glutamate and GABA release, this effect being completely inhibited by the cyclic ADP-ribose specific antagonist 8-NH2-cADPR. Furthermore, addition of NAD+ to astrocyte-neuron co-cultures results in a delayed intracellular calcium transient in neuronal cells, which is strongly but not completely inhibited by glutamate receptor blockers. These data indicate that an astrocyte to-neuron calcium signalling can be triggered by the CD38/cADPR system, which, through the activation of intracellular calcium responses in astrocytes, is in turn responsible for the increased release of neuromodulators from glial cells. PMID- 11483669 TI - Cellular resistance to Evans blue toxicity involves an up-regulation of a phosphate transporter implicated in vesicular glutamate storage. AB - It has recently been suggested that the brain-specific Na+-dependent phosphate inorganic co-transporter (BNPI) is able to support glutamate transport and storage in synaptic vesicles. A procedure for measuring the vesicular pool of glutamate is described and was used to select cell lines according to their glutamate storage capacity. Two cell lines were selected: C6BU-1, with a large intracellular glutamate storage capacity, and NG108-15, devoid of it. Their contents in BNPI mRNA were compared by RT-PCR. We found that both cell lines had BNPI, but in addition C6BU-1 alone expresses the other isoform, DNPI. We also carried out a clonal selection of NG108-15 cells in the presence of the dye Evans blue, a competitive inhibitor of vesicular glutamate transport, very toxic for cells in culture. It was assumed that only those that sequester and eliminate the drug by overexpressing a vesicular glutamate transporter would survive. We found that the NG108-15 clones resistant to Evans blue had an increased storage capacity for glutamate. These cells also up-regulated the BNPI isoform of the phosphate transporter as shown by RT-PCR and northern blot. PMID- 11483670 TI - Regulation of MARCKS and MARCKS-related protein expression in BV-2 microglial cells in response to lipopolysaccharide. AB - Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP) have been implicated in membrane-cytoskeletal events underlying cell adhesion, migration, secretion, and phagocytosis. In BV-2 microglial cells, lipopolysaccharide (LPS) elicited a dose-dependent increase in mRNA of both MRP (sixfold) and MARCKS (threefold) with corresponding increases in [3H]myristoylated and immunoreactive protein levels. LPS also produced significant increases in protein kinase C (PKC)-beta twofold and PKC-epsilon (1.5 fold). Pro-inflammatory cytokines produced by activated microglia (IL-1beta, IL 6, TNF-alpha) did not mimic LPS effects on MARCKS or MRP expression when added individually or in combination. LPS and IFN-gamma produced a synergistic induction of iNOS but not MARCKS or MRP. Induction of MARCKS and MRP by LPS was completely blocked by inhibitors of NF-kappaB (PDTC) and protein tyrosine kinases (herbimycin A), partially blocked by the p38 kinase inhibitor SB203580, and unaffected by the MEK inhibitor PD98059. LPS induction of iNOS was considerably more sensitive to all these inhibitors. The Src kinase inhibitor PP2 had no effect, while the closely related inhibitor PP1 actually increased LPS induction of MARCKS and MRP. Our results suggest that MARCKS and MRP may play an important role in LPS-activated microglia, but are not part of the neuroinflammatory response produced by cytokines. PMID- 11483671 TI - Effect of whole-body (18)F-FDG PET imaging on clinical staging and management of patients with malignant lymphoma. AB - Correct staging is important in selecting the appropriate treatment for lymphoma patients. PET imaging with (18)F-FDG is useful for staging of lymphoma as well as for monitoring of therapy. However, to our knowledge, the clinical impact of PET on staging and management of lymphoma patients has not been reported. METHODS: Standardized questionnaires were mailed to referring physicians asking them whether and how the results of PET imaging had influenced clinical staging and management of the disease in their patients. Management changes, when present, were classified as intermodality (e.g., medical to surgical, surgical to radiation, medical to no treatment) or intramodality (e.g., altered medical, surgical, or radiotherapy approach). RESULTS: The referring physicians returned 52 of 108 questionnaires (48.1%). Physicians indicated that PET led to a change in the clinical stage in 44% of patients: 21% were upstaged and 23% were downstaged. Findings of the PET examination resulted in intermodality changes in management in 42% of patients, in intramodality changes in 10%, and in a combination of the management changes in 10%. Other, not further specified, treatment changes were reported in 6% of patients. PET did not result in any management changes in only 32% of patients. CONCLUSION: This survey-based study of referring physicians indicates that FDG PET has a major impact on the management of lymphoma patients, contributing to changes in clinical stage in 44% and changes in treatment in >60% of cases. PMID- 11483672 TI - Correlation between postoperative 3-[(123)I]iodo-L-alpha-methyltyrosine uptake and survival in patients with gliomas. AB - The aim of this study was to evaluate the prognostic value of SPECT imaging using the amino acid analog 3-[(123)I]iodo-L-alpha-methyltyrosine (IMT) in patients with gliomas. METHODS: One hundred fourteen consecutive patients with newly diagnosed gliomas were examined by IMT SPECT (low-grade glioma, n = 12; anaplastic astrocytoma or oligodendroglioma, n = 46; glioblastoma, n = 56). Seventy-one of these patients had undergone tumor resection 4-6 wk before SPECT imaging (group A). Forty-three patients with unresectable tumors were examined after stereotactic biopsy (group B). IMT uptake at the site of the tumor was assessed visually and quantified relative to a contralateral reference region (IMT uptake ratio). After IMT SPECT, all patients were treated with conformal radiotherapy. The median follow-up time was 27 mo. RESULTS: In group A, focal IMT uptake at the resection site was visible in 52 of 71 patients (73%). Median survival was only 13 mo in these patients, whereas median survival was reached in patients without focal IMT uptake (P = 0.02). Furthermore, the intensity of IMT uptake significantly correlated with survival: patients with an IMT uptake ratio > 1.7 were at a 4.6 times higher risk of death than were patients with a lower IMT uptake (P < 0.001). The IMT uptake ratio remained a significant prognostic factor when age and grading were included in a multivariate model. In contrast, IMT uptake did not correlate with survival in group B (P = 0.95). CONCLUSION: In patients with unresectable high-grade gliomas, IMT uptake appears not to correlate with the biologic aggressiveness of tumor cells. Nevertheless, the clear association between focal IMT uptake after tumor resection and poor survival suggests that IMT is a specific marker for residual tumor tissue. Therefore, IMT SPECT is expected to become a valuable tool for the planning and monitoring of local therapeutic modalities. PMID- 11483673 TI - Brain perfusion spect in juvenile neuro-Behcet's disease. AB - Regional cerebral blood flow was evaluated by (99m)Tc-hexamethylpropyleneamine oxime SPECT in 7 patients (age range, 7--18 y; mean age, 9.1 y) affected with Behcet's disease and signs or symptoms of central nervous system involvement at different times of their clinical history. METHODS: Three patients suffered from seizures, 3 patients were affected with severe persistent headache that was refractory to common analgesic and nonsteroidal antiinflammatory drugs, and 1 patient had recurrent episodes of acute intracranial hypertension. Electroencephalography was performed on all patients, MRI on 5 patients, and CT on 1 patient. Brain SPECT was performed using a high-resolution, brain-dedicated camera. After conventional visual analysis by 2 expert readers, 2 transaxial sections were drawn parallel to the bicommissural line: the first across the thalami and the second across the temporal lobe at the level of the mesiotemporal structures. Cortical regions of interest were drawn automatically on the cortical ribbon on the 2 sections, whereas other regions of interest were drawn by hand around the basal ganglia, the thalami, and the mesiotemporal structures. Asymmetry analysis was then applied, and hypoperfusion was considered when the asymmetry value was >10%. RESULTS: Hypoperfusion was observed in all patients by visual and asymmetry analyses; this finding was localized mainly in the basal ganglia, the thalami, and the temporal cortex, including its mesial portion. Temporal hypoperfusion was found primarily in patients with seizures, and hypoperfusion of deep gray nuclei was found mainly in the other patients. Electroencephalography disclosed brain functional impairment in 5 of 6 patients, where- as MRI showed multiple bilateral white matter lesions in 1 patient suffering from persistent headache. CONCLUSION: As in adults, perfusion SPECT seems to be very sensitive in disclosing brain abnormalities in children and adolescents with Behcet's disease and signs or symptoms of central nervous system involvement, even with negative findings on brain MRI. PMID- 11483674 TI - The utility of a 3-dimensional, large-field-of-view, sodium iodide crystal--based PET scanner in the presurgical evaluation of partial epilepsy. AB - (18)F-FDG PET is an accurate and reliable technique for localizing medically refractory temporal lobe epilepsy, but widespread use has been hindered by limited reimbursement in many countries because of the high cost of traditional PET equipment and radioisotopes. Additionally, the place of FDG PET as a cost effective tool for presurgical evaluation of epilepsy has been questioned because of limited data showing that FDG PET provides localization information incremental to that provided by more established techniques, particularly MRI and ictal electroencephalography (EEG). Three-dimensional (3D), large-field-of-view, sodium iodide crystal-based scanners have lower equipment and running costs and better multiplanar resolution than traditional 2-dimensional bismuth germinate (BGO) systems but have not yet been validated for evaluation of epilepsy. Our purpose was to investigate the localization rate, accuracy, and prognostic value of FDG PET images acquired on a 3D, large-field-of-view, sodium iodide crystal based PET scanner in the presurgical evaluation of intractable partial epilepsy. We also wanted to establish the incremental value of FDG PET over established MRI and ictal EEG techniques. METHODS: Fifty-five patients who were surgical candidates because of medically refractory partial epilepsy were examined. For most of these patients, the lesions had not been clearly localized on conventional assessment. The FDG PET scans were reviewed independently by 2 reviewers who were unaware of the patients' clinical details, ictal EEG findings, and volumetric MRI results, and the FDG PET results were correlated with those of MRI and EEG and with postsurgical outcome. RESULTS: Forty-two patients (76%) had localizing FDG PET images (37 temporal, 5 extratemporal). The ictal EEG recordings were localizing in 66%, and the MRI findings were localizing in 27% (which increased to 35% after the MRI findings were reviewed again after PET). Concordance between the site of the PET localizations and the site of the MRI or EEG localizations was 100%. The PET images were localizing in 63% and 69% of patients with nonlocalizing ictal EEG and MRI findings, respectively. Twenty-one of 24 patients who subsequently underwent epilepsy surgery had localizing FDG PET images; of these 21 patients, 18 (86%) had a class I outcome. Multiple regression analysis showed the FDG PET results to be predictive of postsurgical outcome independently of the MRI findings. CONCLUSION: For intractable partial epilepsy, FDG PET using a 3D, large-field-of-view, sodium iodide crystal-based scanner provided clinically useful localizing information that was at least as accurate as the results reported for traditional BGO-based scanners. The PET images provided prognostically significant localization information incremental to that provided by volumetric MRI and ictal EEG, particularly if 1 of these studies was nonlocalizing. PMID- 11483675 TI - Clinical outcome of patients with previous myocardial infarction and left ventricular dysfunction assessed with myocardial (99m)Tc-MIBI SPECT and (18)F-FDG PET. AB - Myocardial viability was assessed by (99m)Tc-methoxyisobutylisonitrile (MIBI) SPECT and (18)F-FDG PET to evaluate the prognosis and treatment strategy of patients with myocardial infarction (MI) and left ventricular (LV) dysfunction. METHODS: One hundred twenty-three consecutive patients with previous MI and LV dysfunction (LV ejection fraction [EF], 35% +/- 6% [mean +/- SD]) who underwent (99m)Tc-MIBI SPECT and FDG PET were followed-up for 26 +/- 10 mo (mean +/- SD). Distributions of the 2 radiotracers in myocardial segments were classified into 2 patterns: myocardial perfusion-metabolism mismatch (MM) and match (M). LV EF and LV end-diastolic diameter (EDD) were measured by echocardiography at baseline, 3 mo (Pos1), and 6 mo (Pos2) after revascularization. Cardiac death, acute MI, unstable angina, and late revascularization (>3 mo) experienced by the patients during follow-up were defined as cardiac events. RESULTS: Sixty-seven patients underwent revascularization and 56 patients were treated medically. Of the 72 patients with > or =2 MM segments, 42 underwent revascularization (group A1) and 30 were treated medically (group A2). Of the 51 patients with <2 MM segments, 25 underwent revascularization (group B1) and 26 were treated medically (group B2). The 4 groups had similar baseline characteristics and rest LV EF. After revascularization, EF (mean +/- SD) increased in group A1 from 36% +/- 5% to 44% +/- 8% (P < 0.0001) in Pos1 and to 51% +/- 9% (P < 0.0001) in Pos2. EDD (mean +/- SD) decreased from 62 +/- 8 mm to 56 +/- 5 mm (P < 0.001) in Pos1 and to 55 +/- 5 mm (P < 0.001) in Pos2. However, EF and EDD were unchanged in group B1 (P > 0.05). During the follow-up, 22 patients (17.9%) suffered from cardiac events, including 11 cardiac deaths, 4 acute MI, 6 late coronary artery bypass grafting, and 1 unstable angina pectoris. The cardiac event rate in group A2 (50%) was significantly higher than that of groups A1 (2.4%; chi(2) = 23.08; P < 0.0001), B1 (12%; chi(2) = 8.94; P = 0.003), and B2 (11.5%; chi(2) = 9.45; P = 0.002). CONCLUSION: Assessment of myocardial viability using hybrid (99m)Tc-MIBI SPECT and FDG PET can predict the clinical outcome and is helpful to decision making in the treatment strategy of patients with MI and LV dysfunction. Revascularization can improve the LV function and clinical outcome of patients with >2 viable myocardial segments. PMID- 11483676 TI - [1-(11)C]Acetate as a quantitative perfusion tracer in myocardial PET. AB - Our objective was to investigate the properties of [1-(11)C]acetate as a quantitative perfusion tracer for myocardial PET studies. METHODS: We determined the flow dependence of the effective acetate extraction by a comparison with [(13)N]ammonia in 24 patients at rest (n = 8) and under pharmacologic vasodilation (n = 16). Furthermore, we compared the statistical quality of the perfusion values derived with both tracers. Quantification was based on an irreversible 2-compartment model for [(13)N]ammonia and a reversible 1 compartment model for [1-(11)C]acetate. Area-conserving polar maps were used to determine the correlation between the unidirectional uptake parameters of both tracers on a pixel-by-pixel basis for the whole left ventricular myocardium. RESULTS: A fit of a generalized Renkin-Crone formula to the data yielded the unidirectional acetate extraction fraction E(f) = 1 - 0.64e(-1.20/f). An extraction correction based on this formula led to good quantitative agreement of perfusion values derived with [(13)N]ammonia and [1-(11)C]acetate over the whole observed flow range (average difference of flow values, 3%; correlation coefficient, 0.96). This agreement proved the applicability of acetate as a quantitative perfusion tracer even under stress conditions. An analysis of the statistical properties of the parameter estimates showed, moreover, that statistical errors were reduced by a factor of nearly 2 in comparison with ammonia. CONCLUSION: [1-(11)C]acetate allows accurate quantification of myocardial perfusion with PET at rest as well as under stress conditions. The use of acetate leads to distinctly improved statistical accuracy for the perfusion estimates in comparison with ammonia. This accuracy facilitates the generation of reliable parametric polar maps, which are especially useful for clinical application of myocardial perfusion quantification. PMID- 11483677 TI - What is the best approach to quantify myocardial blood flow with pet? PMID- 11483678 TI - Diagnostic performance of an expert system for the interpretation of myocardial perfusion SPECT studies. AB - An expert system (PERFEX) developed for the computer-assisted interpretation of myocardial perfusion SPECT studies is now becoming widely available. To date, a systematic validation of the diagnostic performance of this expert system for the interpretation of myocardial perfusion SPECT studies has not been reported. METHODS: To validate PERFEX's ability to detect and locate coronary artery disease (CAD), we analyzed 655 stress/rest myocardial perfusion prospective SPECT studies in patients who also underwent coronary angiography. The patient population comprised CAD patients (n = 480) and healthy volunteers (n = 175) (449 men, 206 women). Data from 461 other patient studies were used to implement and refine 253 heuristic rules that best correlated the presence and location of left ventricular myocardial perfusion defects on SPECT studies with angiographically detected CAD and with human expert visual interpretations. Myocardial perfusion defects were automatically identified as segments with counts below sex-matched normal limits. PERFEX uses the certainty of the location, size, shape, and reversibility of the perfusion defects to infer the certainty of the presence and location of CAD. The visual interpretations of tomograms and polar maps, vessel stenosis from coronary angiography, and PERFEX interpretations were all accessed automatically from databases and were used to automatically generate comparisons between diagnostic approaches. RESULTS: Using the physician's reading as a gold standard, PERFEX's sensitivity and specificity levels for detection and localization of disease were, respectively, 83% and 73% for CAD, 76% and 66% for the left anterior descending artery, 90% and 70% for the left circumflex artery, and 74% and 79% for the right coronary artery. These results were extracted from a receiver operating characteristic curve using the average optimal input certainty factor. CONCLUSION: This study shows that the diagnostic performance of PERFEX for interpreting myocardial perfusion SPECT studies is comparable with that of nuclear medicine experts in detecting and locating CAD. PMID- 11483679 TI - Use of artificial intelligence in cardiac imaging. PMID- 11483680 TI - Comparison of PET oxygen extraction fraction methods for the prediction of stroke risk. AB - PET measurement of increased oxygen extraction fraction (OEF) identifies patients at high risk for subsequent stroke. OEF methodology remains controversial. In this study we compare the sensitivity and specificity of absolute OEF measurements with ipsilateral-to-contralateral ratios of absolute OEF and count based OEF estimates. METHODS: Multivariate analyses of OEF methods were performed using data from patients with symptomatic carotid artery occlusion (n = 68). Outcome and receiver operating characteristic (ROC) curve analyses were performed. RESULTS: All 3 methods were predictive of stroke risk in univariate analysis. Only the count-based method remained significant in multivariate analysis. The area under the ROC curve was greatest for the count-based ratio: 0.815 versus 0.769 (absolute) and 0.737 (ratios of absolute). CONCLUSION: All 3 methods are predictive of stroke risk in patients with unilateral carotid artery occlusion. ROC curve analysis is useful for selecting optimal thresholds for maximal sensitivity and specificity. PMID- 11483681 TI - Radioguided sentinel lymph node biopsy in breast cancer surgery. AB - The concept of sentinel lymph node biopsy in breast cancer surgery relates to the fact that the tumor drains in a logical way through the lymphatic system, from the first to upper levels. Therefore, the first lymph node met (the sentinel node) will most likely be the first to be affected by metastasis, and a negative sentinel node makes it highly unlikely that other nodes are affected. Because axillary node dissection does not improve prognosis of patients with breast cancer (being important only to stage the axilla), sentinel lymph node biopsy might replace complete axillary dissection to stage the axilla in clinically N0 patients. Sentinel lymph node biopsy would represent a significant advantage as a minimally invasive procedure, considering that, after surgery, about 70% of patients are found to be free from metastatic disease, yet axillary node dissection can lead to significant morbidity. Furthermore, histologic sampling errors can be reduced if a single (sentinel) node is assessed extensively rather than few histologic sections in a high number of lymph nodes per patient. Although the pattern of lymph drainage from breast cancer can be variable, the mammary gland and the overlying skin can be considered as a biologic unit in which lymphatics tend to follow the vasculature. Therefore, considering that tumor lymphatics are disorganized and relatively ineffective, subdermal and peritumoral injection of small aliquots of radiotracer is preferred to intratumoral administration. (99m)Tc-labeled colloids with most of the particles in the 100- to 200-nm size range would be ideal for radioguided sentinel node biopsy in breast cancer. Lymphoscintigraphy is an essential part of radioguided sentinel lymph node biopsy because images are used to direct the surgeon to the site of the node. The sentinel lymph node should have a significantly higher count than that of background (at least 10:1 intraoperatively). After removal of the sentinel node, the axilla must be reexamined to ensure that all radioactive sites are identified and removed for analysis. The sentinel lymph node should be processed for intraoperative frozen section examination in its entirety, based on conventional histopathology and, when needed, immune staining with anticytokeratin antibody. The success rate of radioguidance in localizing the sentinel lymph node in breast cancer surgery is about 94%--97% in institutions where a high number of procedures are performed and approaches 99% when combined with the vital blue dye technique. At present, there is no definite evidence that negative sentinel lymph node biopsy is invariably correlated with negative axillary status, except perhaps for T1a-b breast cancers, with a size of < or =1 cm. Randomized clinical trials should elucidate the impact of avoiding axillary node dissection on patients with a negative sentinel lymph node on the long-term clinical outcome of patients. PMID- 11483682 TI - Cellular dose conversion factors for alpha-particle--emitting radionuclides of interest in radionuclide therapy. AB - alpha-Particle--emitting radionuclides are of increasing interest in radionuclide therapy. The decay scheme of alpha-emitting radionuclides typically includes a chain of unstable progeny. It is generally assumed that alpha-particle emission by the parent radionuclide will break the chemical bond with its carrier molecule and that the resulting daughter atom will no longer be associated with the carrier molecule. If the daughter is very short lived, it will not have enough time to be carried any significant distance from the site of parent decay and a cellular, absorbed dose estimate must consider the energy deposited by the daughter as well as the parent. Depending on the site of parent decay and the expected removal rate of daughter atoms from this site, the contribution of emissions from longer-lived daughters may also be warranted. In this study, dose conversion factors (DCFs) for cellular dimensions that incorporate the fate of daughter radionuclides were derived for (225)Ac, (213)Bi, (211)At, and (223)Ra, the alpha-particle--emitting radionuclides of interest in radionuclide therapy. METHODS: The dose contribution of daughter radionuclides at the site of parent decay was made dependent on a cutoff time parameter, which was used to estimate the fraction of daughter decays expected at the site of parent decay. Previously tabulated S values (cell-surface to nucleus and cell-surface to cell) for each daughter in the decay scheme were scaled by this fraction and a sum over all daughters was performed to yield a cutoff time--dependent set of corresponding DCF values for each radionuclide. RESULTS: DCF values for the absorbed dose to the nuclear or cellular volume from cell-surface decays are presented as a function of the cutoff time for 4 different cellular and nuclear dimensions. CONCLUSION: In contrast to the cellular S values that account only for parent decay, the DCF values provided in this study make it possible to easily include the contribution of daughter decays in cellular alpha-particle emitter dose calculations. PMID- 11483683 TI - alpha-Particle emitters in radioimmunotherapy: new and welcome challenges to medical internal dosimetry. PMID- 11483684 TI - Human pharmacokinetic and dosimetry studies of [(18)F]FHBG: a reporter probe for imaging herpes simplex virus type-1 thymidine kinase reporter gene expression. AB - 9-[4-[(18)F]fluoro-3-(hydroxymethyl)butyl]guanine ([(18)F]FHBG) has been used as a reporter probe to image expression of herpes simplex virus type-1 thymidine kinase (HSV1-tk) reporter gene in living animals. Our aim was to study the kinetics, biodistribution, stability, dosimetry, and safety of [(18)F]FHBG in healthy human volunteers, preparatory to imaging patients undergoing HSV1-tk gene therapy. METHODS: [(18)F]FHBG was synthesized with a specific activity of 37,000- 444,000 GBq/mmol and a radiochemical purity > 99%. Ten healthy volunteers consented to participate in the study. A transmission scan was obtained before bolus injection of 70.3--229.4 MBq [(18)F]FHBG into a hand vein, followed by dynamic PET imaging with 4 consecutive emission scans. Warmed hand-vein blood was withdrawn at various times after injection for blood time--activity measurements. Electrocardiography, blood pressure, and blood and urine pharmacologic parameters were measured before and after injection of the [(18)F]FHBG tracer (n = 5). The stability of [(18)F]FHBG in the urine was analyzed. Attenuation-corrected images were reconstructed using the ordered-subsets expectation maximization algorithm. Image region-of-interest time-activity data were used with the MIRD program to estimate absorbed radiation dosages. RESULTS: [(18)F]FHBG had rapid blood clearance; only 8.42% +/- 4.76% (mean +/- SD) of the peak blood activity remained at approximately 30 min. The average ratio of plasma activity to whole-blood activity during the study was 0.91 +/- 0.04. Penetration of [(18)F]FHBG across the blood-brain barrier was not observed. The primary routes of clearance were renal and hepatobiliary. High activities were observed in the bladder, gut, liver, and kidneys, but <0.0002% of the injected dose per gram was observed in other tissues. In the urine, 83% of activity 180 min after injection was stable [(18)F]FHBG. Blood and urine pharmacologic parameters did not change significantly after injection of the [(18)F]FHBG tracer. The bladder absorbed the highest radiation dose. CONCLUSION: [(18)F]FHBG has the desirable in vivo characteristics of stability, rapid blood clearance, low background signal, biosafety, and acceptable radiation dosimetry in humans. This study forms the foundation for using [(18)F]FHBG in applications to monitor HSV1-tk reporter gene expression. PMID- 11483685 TI - Progress in gene therapy: seeing is believing. PMID- 11483686 TI - Pattern of uptake and excretion of (18)F-FDG in the lactating breast. AB - Excretion of radiopharmaceuticals into breast milk poses a potential risk to infants and clear recommendations regarding interruption times are required. There are few data available regarding the impact of (18)F-FDG on this issue. With increasing use of PET for oncologic imaging and its potential advantages to nursing mothers because of its short physical half-life compared with other commonly used tumor imaging agents such as (67)Ga and (201)Tl, evaluation of the excretion pattern of this agent in breast milk is important. METHODS: We have evaluated the uptake of FDG in the breasts in 7 women, 6 of whom were lactating and 1 of whom was in early postpartum but had not commenced breast-feeding. Milk samples were obtained from 4 of the lactating women, including serial samples from 1. RESULTS: Significantly increased breast uptake was identified in all lactating breasts but not in 1 breast consistently refused by the nursing infant or in the woman who had not begun breast-feeding after delivery of her child. No qualitative change or semiquantitative estimate of radiotracer uptake in the breast was seen after expression of breast milk. Decay-corrected activity measurable in breast milk ranged from 5.54 to 19.3 Bq/mL/MBq injected. Using a standard model of breast-feeding, the calculated maximum cumulative dose to the infant, 0.085 mSv with no interruption of breast-feeding, is well below the recommended limit of 1 mSv. CONCLUSION: High uptake of FDG in the lactating breast appears to be related to suckling. There is, however, little secretion of activity into breast milk. Accordingly, a higher radiation dose is received by the infant from close contact with the breast than from ingestion of radioactive milk. PMID- 11483687 TI - Preferential labeling of glial and meningial brain tumors with [2-(14)C]acetate. AB - Acetate is preferentially transported into and metabolized by astrocytes, rather than synaptosomes or neurons, and labeled acetate is used as a glial reporter molecule to assess glial metabolism and glial-neuronal interactions. Because monocarboxylic acid transporter specificity might confer a phenotype to help localize, detect, and characterize brain tumors of glial origin, use of [2 (14)C]acetate and [(14)C]deoxyglucose (a glucose analog metabolized by all brain cells) was compared in rat and human brain tumors. METHODS: Cultured C6 glioma or U-373 glioblastoma/astrocytoma tumor cells were injected into the caudate nucleus of anesthetized CDF Fisher rats; 2--3 wk later, an intravenous pulse of [2 (14)C]acetate or [(14)C]deoxyglucose was given, and timed blood samples were drawn during the 5- or 45-min experiment, respectively. Local (14)C levels in the brain were assayed by quantitative autoradiography, and acetate uptake or glucose use was calculated. Uptake and metabolism of the [(14)C]acetate was also assayed in C6 glioma and human surgical tumor samples in vitro. RESULTS: [(14)C]Acetate uptake into rat brain C6 tumors was 9.9 +/- 2.1 mL/100 g/min, compared with 3.9 +/- 1.0 mL/100 g/min in contralateral tissue (n = 6; P < 0.001), and was much higher than that into other brain structures (e.g., 5:1 for white matter and 2:1 for cortical gray matter). Glucose use in C6 tumors was 111 +/- 34 micromol/100 g/min, versus 81 +/- 5 micromol/100 g/min in contralateral tissue (n = 6; P = 0.08); no left-right differences in glucose use or acetate uptake were seen in other brain structures. The tumor-to-contralateral-tissue ratio for acetate (2.3 +/- 0.3) exceeded that for deoxyglucose (1.4 +/- 0.5) (P < 0.05), indicating that acetate is a sensitive C6 glioma marker. [(14)C]Acetate uptake also demarcated a few 3-wk-old C6 tumors that had unlabeled necrotic cores. U-373 tumors were smaller than C6 tumors in rat brain and were detected equally well with [(14)C]acetate and [(14)C]deoxyglucose. In vitro uptake of [(14)C]acetate into human glioblastoma or meningioma tumors was higher than uptake into pituitary adenoma. Rat C6 and human tumors with high uptake metabolized acetate to acidic compounds and amino acids. CONCLUSION: Tumor imaging with radiolabeled acetate can help to localize and classify brain tumors. Transporter and metabolic substrate specificity are traits that can be exploited further for in vivo imaging of brain glial tumors. PMID- 11483688 TI - Relative therapeutic efficacy of (125)I- and (131)I-labeled monoclonal antibody A33 in a human colon cancer xenograft. AB - A33, a monoclonal antibody that targets colon carcinomas, was labeled with (125)I or (131)I and the relative therapeutic efficacy of the 2 radiolabeled species was compared in a human colon cancer xenograft system. METHODS: Nude mice bearing human SW1222 colon carcinoma xenografts were administered escalating activities of (125)I-A33 (9.25-148 MBq) or (131)I-A33 (0.925-18.5 MBq), (125)I- and (131)I labeled control antibodies, unlabeled antibody, or no antibody. The effects of treatment were assessed using the endpoints of tumor growth delay and cure. RESULTS: Tumor growth delay increased with administered activity for all radiolabeled antibodies. Approximately 4.5 times more activity was required for (125)I-A33 to produce therapeutic effects that were equivalent to those of (131)I A33. This ratio was approximately 7 for a nonspecific, noninternalizing isotype matched, radiolabeled control antibody. Unlabeled A33 antibody had no effect on tumor growth. Approximately 10 times more activity of (125)I-A33 produced toxicity similar to that of (131)I-A33, and this ratio fell to approximately 6 for radiolabeled control antibody. CONCLUSION: Treatment with (125)I-A33 resulted in a relative therapeutic gain of approximately 2 compared with (131)I-A33 in this experimental system. PMID- 11483689 TI - (99m)Tc-interleukin-8 for imaging acute osteomyelitis. AB - Early and accurate diagnosis of osteomyelitis remains a clinical problem. Acute osteomyelitis often occurs in infants and most often is located in the long bones. Radiologic images show changes only in advanced stages of disease. Scintigraphic imaging with (99m)Tc-methylene diphosphonate (MDP), or bone scanning, is much more sensitive in detecting acute osteomyelitis but lacks specificity. We evaluated the performance of (99m)Tc-interleukin-8 (IL-8) in an experimental model of acute osteomyelitis. METHODS: Acute pyogenic osteomyelitis was induced in 10 rabbits by inserting sodium morrhuate and Staphylococcus aureus into the medullary cavity of the right femur. The cavity was closed with liquid cement. A sham operation was performed on the left femur. Routine radiographs were obtained just before scintigraphy. Ten days after surgery, the rabbits were divided into 2 groups of 5 animals, received an injection of either 18.5 MBq (111)In-granulocytes or 18.5 MBq (67)Ga-citrate, and were imaged both 24 h after injection and 48 h after injection. On day 12, the rabbits received either 18.5 MBq (99m)Tc-MDP or 18.5 MBq (99m)Tc-IL-8, and serial images were acquired at 0, 1, 2, 4, 8, 12, and 24 h after injection. Uptake in the infected femur was determined by drawing regions of interest. Ratios of infected femur (target) to sham-operated femur (background) (T/Bs) were calculated. After the final images were obtained, the rabbits were killed and the right femur was dissected and analyzed for microbiologic and histopathologic evidence of osteomyelitis. RESULTS: Acute osteomyelitis developed in 8 of 10 rabbits. All imaging agents correctly detected the acute osteomyelitis in these animals. The extent of infection was optimally visualized with (67)Ga-citrate and delayed bone scanning, whereas diaphyseal photopenia was noted with both (99m)Tc-IL-8 and (111)In granulocytes. In 1 rabbit with osteomyelitis, imaging results were falsely negative with (111)In-granulocytes and falsely positive with (99m)Tc-MDP. Quantitative analysis of the images revealed that the uptake in the infected region was highest with (67)Ga-citrate (4.9 +/- 0.8 percentage injected dose [%ID]) and (99m)Tc-MDP (4.7 +/- 0.7 %ID), whereas the uptake in the infected area was significantly lower with (99m)Tc-IL-8 (2.2 +/- 0.2 %ID) and (111)In granulocytes (0.8 +/- 0.2 %ID) (P < 0.0042). In contrast, the T/Bs were significantly higher for (99m)Tc-IL-8 (T/B, 6.2 +/- 0.3 at 4 h after injection) than for (67)Ga-citrate, (99m)Tc-MDP, and (111)In-granulocytes, which had ratios of 1.5 +/- 0.4, 1.9 +/- 0.2, and 1.4 +/- 0.1, respectively (P < 0.0001). Radiography correctly revealed acute osteomyelitis in only 2 of 8 rabbits. CONCLUSION: In this rabbit model of osteomyelitis, (99m)Tc-IL-8 clearly revealed the osteomyelitic lesion. Although the absolute uptake in the osteomyelitic area was significantly lower than that obtained with (99m)Tc-MDP and (67)Ga-citrate, the T/Bs were significantly higher for (99m)Tc-IL-8 because of fast background clearance. The ease of preparation, good image quality, and lower radiation burden suggest that (99m)Tc-IL-8 may be a suitable imaging agent for the scintigraphic evaluation of acute osteomyelitis. PMID- 11483690 TI - Quantification of [(18)F]FDG uptake in the normal liver using dynamic PET: impact and modeling of the dual hepatic blood supply. AB - For quantification of hepatic [(18)F]FDG uptake, the dual blood supply to the liver must be considered. In contrast to the arterial input, however, the portal venous blood supply to the liver cannot be monitored directly by PET because of the inaccessibility of the portal vein on PET scans. In this study, we investigated whether the dual hepatic input can be predicted from the measurable arterial input. Moreover, we assessed the effect of different input models on the rate constants of the standard 3-compartment model describing regional uptake of FDG. METHODS: Dynamic FDG PET scanning was performed on 5 foxhounds. Activity concentrations in blood from the aorta and the portal vein were measured simultaneously using external circuits. After image reconstruction, time- activity courses were determined from the aorta and the liver. The venous input was approximated by convolving the arterial input with a notional system function describing the dispersion of the arterial input on its way through the gastrointestinal tract. On the basis of these data, 5 different hepatic input models, which pertain to a single-input as well as a dual-input scenario, were statistically compared with regard to the adequacy of the model fits to liver data and to differences in the estimated rate constants. RESULTS: Portal venous input to the liver could be approximated by convolving the arterial input function with a system function. From this function, a mean transit time of 25 s was computed for FDG to pass through the gastrointestinal tract. According to the statistical analysis, dual-input models were superior to their single-input counterparts. However, differences in the rate constants estimated for the 5 input models were in the same order as interindividual variations within the different model groups. For the dephosphorylation rate constant, a consistent value of 0.05 +/- 0.01 min(-1) was found. CONCLUSION: Dual-input models proved to be superior to single-input models with respect to the adequacy of FDG model fits to normal liver data. However, the hepatic blood supply may be approximated by the arterial input function as well, especially for the evaluation of liver lesions mainly fed by the hepatic artery. PMID- 11483691 TI - In vivo labeling of endothelin receptors with [(11)C]L-753,037: studies in mice and a dog. AB - Endothelin (ET) is a potent mammalian vasoconstrictive peptide and a pressor agent. Its 3 isoforms, ET-1, ET-2, and ET-3, mediate several physiologic actions in several organ systems, binding to 2 major receptor subtypes: ET(A) and ET(B). This study was undertaken to evaluate [(11)C]L-753,037 [(+)-(5S,6R,7R)-2-butyl-7 [2-((2S)-2-carboxy-propyl)-4-methoxyphenyl]-5-(3,4 methylenedioxyphenyl)cyclopenteno [1,2-beta]pyridine-6-carboxylate), a new mixed ET receptor A and B antagonist, as a tracer for in vivo labeling of ET receptors in mice and a dog. METHODS: [(11)C]L-753,037 was synthesized, purified, and formulated from a normethyl precursor, L-843,974, and [(11)C]H(3)I. The tracer was studied for its in vivo kinetics, biodistribution, and ET receptor binding characteristics in mice. In the dog, PET imaging was performed to evaluate binding of [(11)C]L-753,037 to ET receptors in the heart. Specificity of binding was studied in the heart with the selective ET(A) antagonist L-753,164. RESULTS: Kinetic studies in mice showed highest tracer uptake at 5 min after injection in liver (25.0 percentage injected dose per gram [%ID/g]), kidneys (18.7 %ID/g), lungs (15.2 %ID/g), and heart (5.6 %ID/g). Initial high uptake in liver, lungs, and kidneys was followed by rapid washout during the next 10 min and a very slow clearance during the time of observation (2 h after injection). By contrast, the radioactivity in the heart remained constant over 2 h. Administration of both ET(A) (L-753,164) and mixed ET(A)/ET(B) (L-753,137) receptor antagonists resulted in dose-dependent inhibition of [(11)C]L-753,037 binding in mouse heart, lungs, and kidneys but not in the liver. Radioactivity in the brain was very low, indicating that the tracer does not cross the blood-brain barrier. In the dog, a dynamic PET study of the heart showed high tracer accumulation at 55-95 min after injection. Injection of L-753,164 at 30 min before [(11)C]L-753,037 administration led to a significant reduction in tracer binding. [(11)C]methyl triphenyl phosphonium was used as a tracer for reference images of the dog heart muscle. CONCLUSION: The results suggest that [(11)C]L-753,037 binds to ET receptors in vivo and is, therefore, a promising candidate for investigation of these receptors and their occupancy by ET receptor antagonists using PET. PMID- 11483692 TI - PET imaging of (86)Y-labeled anti-Lewis Y monoclonal antibodies in a nude mouse model: comparison between (86)Y and (111)In radiolabels. AB - Absorbed doses in (90)Y radioimmunotherapy are usually estimated by extrapolating from (111)In imaging data. PET using (86)Y (beta(+) 33%; half-life, 14.7 h) as a surrogate radiolabel could be a more accurate alternative. The aim of this study was to evaluate an (86)Y-labeled monoclonal antibody (mAb) as a PET imaging agent and to compare the biodistribution of (86)Y- and (111)In-labeled mAb. METHODS: The humanized anti-Lewis Y mAb hu3S193 was labeled with (111)In or (86)Y through CHX-A"-diethylenetriaminepentaacetic acid chelation. In vitro cell binding and cellular retention of radiolabeled hu3S193 were evaluated using HCT-15 colon carcinoma cells, a cell line expressing Lewis Y. Nude mice bearing HCT-15 xenografts were injected with (86)Y-hu3S193 or (111)In-hu3S193. The biodistribution was studied by measurements of dissected tissues as well as by PET and planar imaging. RESULTS: The overall radiochemical yield in hu3S193 labeling and purification was 42% +/- 2% (n = 2) and 76% +/- 3% (n = 6) for (86)Y and (111)In, respectively. Both radioimmunoconjugates specifically bound to HCT 15 cells. When cellular retention of hu3S193 was studied using (111)In-hu3S193, 80% of initially cell-bound (111)In activity was released into the medium as high molecular-weight compounds within 8 h. When coadministered, in vivo tumor uptake of (86)Y-hu3S193 and (111)In-hu3S193 reached maximum values of 30 +/- 6 and 29 +/ 6 percentage injected dose per gram and tumor sites were easily identifiable by PET and planar imaging, respectively. CONCLUSION: At 2 d after injection of (111)In-hu3S193 and (86)Y-hu3S193 radioimmunoconjugates, the uptake of (111)In and (86)Y activity was generally similar in most tissues. After 4 d, however, the concentration of (86)Y activity was significantly higher in several tissues, including tumor and bone tissue. Accordingly, the quantitative information offered by PET, combined with the presumably identical biodistribution of (86)Y and (90)Y radiolabels, should enable more accurate absorbed dose estimates in (90)Y radioimmunotherapy. PMID- 11483693 TI - Comprehensive ventilation/perfusion SPECT. AB - Lung scintigraphy is the primary tool for diagnostics of pulmonary embolism. A perfusion study is often complemented by a ventilation study. Intermediate probability scans are frequent. Our goal was to develop a fast method for tomographic ventilation and perfusion scintigraphy to improve the diagnostic value of lung scintigraphy. METHODS: SPECT was performed with a dual-head gamma camera. Acquisition parameters were determined using a thorax phantom. Ventilation tomography after inhalation of 30 MBq (99m)Tc diethylenetriaminepentaacetate (DTPA) aerosol was, without patient movement, followed by perfusion tomography after an intravenous injection of 100 MBq (99m)Tc-labeled macroaggregated albumin (MAA). Total SPECT acquisition time was 20 min. (99m)Tc-DTPA clearance, calculated from initial and final SPECT projections, was used for correction of the ventilation projection set before iterative reconstruction of ventilation and perfusion. The ventilation background was subtracted from the perfusion tomograms. A normalized ventilation/perfusion quotient (V/P quotient) image set was calculated. The method was evaluated on a trial group of 15 patients. RESULTS: Ventilation and perfusion images had adequate quality and showed ventilation/perfusion (V/Q quotient) relationships more clearly than did planar images. Frontal and sagittal slices were superior to planar scintigraphy for characterization of embolized areas. The V/Q quotient was supportive, particularly in the patients with chronic obstructive pulmonary disease. CONCLUSION: Fast, high-quality, ventilation/perfusion SPECT with standard isotopes doses is feasible and may contribute to higher objectivity in evaluating lung embolism as well as other lung diseases. The costs for the procedure and patient care until diagnosis are low because of the comprehensive system for the study and, particularly, the short time for its completion. PMID- 11483694 TI - A tabulated summary of the FDG PET literature. PMID- 11483695 TI - Sensory-motor integration in human motor cortex at the pre-motoneurone level: beyond the age of simple MEP measurements. PMID- 11483696 TI - Compartmental neurodegeneration and synaptic plasticity in the Wld(s) mutant mouse. AB - This review focuses on recent developments in our understanding of neurodegeneration at the mammalian neuromuscular junction. We provide evidence to support a hypothesis of compartmental neurodegeneration, whereby synaptic degeneration occurs by a separate, distinct mechanism from cell body and axonal degeneration. Studies of the spontaneous mutant Wld(s) mouse, in which Wallerian degeneration is characteristically slow, provide key evidence in support of this hypothesis. Some features of synaptic degeneration in the absence of Wallerian degeneration resemble synapse elimination in neonatal muscle. This and other forms of synaptic plasticity may be accessible to further investigations, exploiting advantages afforded by the Wld(s) mutant, or transgenic mice that express the Wld(s) gene. PMID- 11483697 TI - The mechanism of phenylephrine-mediated [Ca(2+)](i) oscillations underlying tonic contraction in the rabbit inferior vena cava. AB - 1. We characterized the mechanisms in vascular smooth muscle cells (VSMCs) that produce asynchronous, wave-like Ca(2+) oscillations in response to phenylephrine (PE). Confocal imaging was used to observe [Ca(2+)](i) in individual VSMCs of intact inferior vena cava (IVC) from rabbits. 2. It was found that the Ca(2+) waves were initiated by Ca(2+) release from the sarcoplasmic reticulum (SR) via inositol 1,4,5-trisphosphate-sensitive SR Ca(2+) release channels (IP(3)R channels) and that refilling of the SR Ca(2+) store through the sarcoplasmic endoplasmic reticulum Ca(2+)-ATPase (SERCA) was required for maintained generation of the repetitive Ca(2+) waves. 3. Blockade of L-type voltage-gated Ca(2+) channels (L-type VGCCs) with nifedipine reduced the frequency of PE stimulated [Ca(2+)](i) oscillations, while additional blockade of receptor operated channels/store-operated channels (ROCs/SOCs) with SKF96365 abolished the remaining oscillations. Parallel force measurements showed that nifedipine inhibited PE-induced tonic contraction by 27 % while SKF96365 abolished it. This indicates that stimulated Ca(2+) entry refills the SR to support the recurrent waves of SR Ca(2+) release and that both L-type VGCCs and ROCs/SOCs contribute to this process. 4. Application of the Na(+)-Ca(2+) exchanger (NCX) inhibitors 2',4' dichlorobenzamil (forward- and reverse-mode inhibitor) and KB-R7943 (reverse-mode inhibitor) completely abolished the nifedipine-resistant component of [Ca(2+)](i) oscillations and markedly reduced PE-induced tone. 5. Thus, we conclude that each Ca(2+) wave depends on initial SR Ca(2+) release via IP(3)R channels followed by SR Ca(2+) refilling through SERCA. Na(+) entry through ROCs/SOCs facilitates Ca(2+) entry through the NCX operating in the reverse mode, which refills the SR and maintains PE-induced [Ca(2+)](i) oscillations. In addition some Ca(2+) entry through L-type VGCCs and ROCs/SOCs serves to modulate the frequency of the oscillations and the magnitude of force development. PMID- 11483698 TI - Dihydroxyeicosatrienoic acids are potent activators of Ca(2+)-activated K(+) channels in isolated rat coronary arterial myocytes. AB - 1. Dihydroxyeicosatrienoic acids (DHETs), which are metabolites of arachidonic acid (AA) and epoxyeicosatrienoic acids (EETs), have been identified as highly potent endogenous vasodilators, but the mechanisms by which DHETs induce relaxation of vascular smooth muscle are unknown. Using inside-out patch clamp techniques, we examined the effects of DHETs on the large conductance Ca(2+) activated K(+) (BK) channels in smooth muscle cells from rat small coronary arteries (150-300 microM diameter). 2. 11,12-DHET potently activated BK channels with an EC(50) of 1.87 +/- 0.57 nM (n = 5). Moreover, the three other regioisomers 5,6-, 8,9- and 14,15-DHET were equipotent with 11,12-DHET in activating BK channels. The efficacy of 11,12-DHET in opening BK channels was much greater than that of its immediate precursor 11,12-EET. In contrast, AA did not significantly affect BK channel activity. 3. The voltage dependence of BK channels was dramatically modulated by 11,12-DHET. With physiological concentrations of cytoplasmic Ca(2+) (200 nM), the voltage at which the channel open probability was half-maximal (V(1/2)) was shifted from a baseline of 115.6 +/- 6.5 mV to 95.0 +/- 10.1 mV with 5 nM 11,12-DHET, and to 60.0 +/- 8.4 mV with 50 nM 11,12-DHET. 4. 11,12-DHET also enhanced the sensitivity of BK channels to Ca(2+) but did not activate the channels in the absence of Ca(2+). 11,12-DHET (50 nM) reduced the Ca(2+) EC(50) of BK channels from a baseline of 1.02 +/- 0.07 microM to 0.42 +/- 0.11 microM. 5. Single channel kinetic analysis indicated that 11,12-DHET did not alter BK channel conductance but did reduce the first latency of BK channel openings in response to a voltage step. 11,12-DHET dose-dependently increased the open dwell times, abbreviated the closed dwell times, and decreased the transition rates from open to closed states. 6. We conclude that DHETs hyperpolarize vascular smooth muscle cells through modulation of the BK channel gating behaviour, and by enhancing the channel sensitivities to Ca(2+) and voltage. Hence, like EETs, DHETs may function as endothelium-derived hyperpolarizing factors. PMID- 11483700 TI - Expression and function of native potassium channel [K(V)alpha1] subunits in terminal arterioles of rabbit. AB - 1. In this study we investigated the expression and function of the K(V)alpha1 subfamily of voltage-gated K(+) channels in terminal arterioles from rabbit cerebral circulation. 2. K(+) current was measured from smooth muscle cells within intact freshly isolated arteriolar fragments. Current activated on depolarisation positive of about -45 mV and a large fraction of this current was blocked by 3,4-diaminopyridine (3,4-DAP) or 4-aminopyridine (4-AP), inhibitors of K(V) channels. Expression of cRNA encoding K(V)1.6 in Xenopus oocytes also generated a 4-AP-sensitive K(+) current with a threshold for activation near -45 mV. 3. Immunofluorescence labelling revealed K(V)1.2 to be specifically localised to endothelial cells, and K(V)1.5 and K(V)1.6 to plasma membranes of smooth muscle cells. 4. K(V) channel current in arteriolar fragments was blocked by correolide (which is specific for the K(V)alpha1 family of K(V) channels) but was resistant to recombinant agitoxin-2 (rAgTX2; which inhibits K(V)1.6 but not K(V)1.5). Heterologously expressed K(V)2.1 was resistant to correolide, and K(V)1.6 was blocked by rAgTX2. 5. Arterioles that were mildly preconstricted and depolarised by 0.1-0.3 nM endothelin-1 constricted further in response to 3,4 DAP, 4-AP or correolide, but not to rAgTX2. 6. We suggest that K(V)alpha1 channels are expressed in smooth muscle cells of terminal arterioles, underlie a major part of the voltage-dependent K(+) current, and have a physiological function to oppose vasoconstriction. K(V)alpha1 complexes without K(V)1.5 appear to be uncommon. PMID- 11483699 TI - Direct measurement of single-channel Ca(2+) currents in bullfrog hair cells reveals two distinct channel subtypes. AB - 1. To confer their acute sensitivity to mechanical stimuli, hair cells employ Ca(2+) ions to mediate sharp electrical tuning and neurotransmitter release. We examined the diversity and properties of voltage-gated Ca(2+) channels in bullfrog saccular hair cells by means of perforated and cell-attached patch-clamp techniques. Whole-cell Ca(2+) current records provided hints that hair cells express L-type as well as dihydropyridine-insensitive Ca(2+) currents. 2. Single Ca(2+) channel records confirmed the presence of L-type channels, and a distinct Ca(2+) channel, which has sensitivity towards omega-conotoxin GVIA. Despite its sensitivity towards omega-conotoxin GVIA, the non-L-type channel cannot necessarily be considered as an N-type channel because of its distinct voltage dependent gating properties. 3. Using 65 mM Ca(2+) as the charge carrier, the L type channels were recruited at about -40 mV and showed a single-channel conductance of 13 pS. Under similar recording conditions, the non-L-type channels were activated at approximately -60 mV and had a single-channel conductance of approximately 16 pS. 4. The non-L-type channel exhibited at least two fast open time constants (tau(o) = 0.2 and 5 ms). In contrast, the L-type channels showed long openings (tau(o) = approximately 23 ms) that were enhanced by Bay K 8644, in addition to the brief openings (tau(o) = 0.3 and 10 ms). 5. The number of functional channels observed in patches of similar sizes suggests that Ca(2+) channels are expressed singly, in low-density clusters (2-15 channels) and in high-density clusters (20-80 channels). Co-localization of the two channel subtypes was observed in patches containing low-density clusters, but was rare in patches containing high-density clusters. 6. Finally, we confirmed the existence of two distinct Ca(2+) channel subtypes by using immunoblot and immunohistochemical techniques. PMID- 11483701 TI - Ca(2+) influx in the endothelial cells is required for the bradykinin-induced endothelium-dependent contraction in the porcine interlobar renal artery. AB - 1. To determine the mechanism of bradykinin-induced production of endothelium derived contracting factors, we monitored the changes in cytosolic Ca(2+) concentration ([Ca(2+)](i)) in in situ endothelial cells in porcine aortic valvular strips and the changes in [Ca(2+)](i) of smooth muscle cells and force in porcine interlobar renal arterial strips using front-surface fluorometry of fura-2. 2. In the presence of N(omega)-nitro-L-arginine methyl ester, bradykinin caused an endothelium-dependent transient elevation of [Ca(2+)](i) and contraction in smooth muscle in the interlobar renal artery. This contraction was completely inhibited by a prostaglandin H(2)/thromboxane A(2) receptor antagonist. 3. In the absence of extracellular Ca(2+), bradykinin failed to induce contraction. However, replenishing extracellular Ca(2+) to 0.75 mM and higher induced an instantaneous contraction. However, replenishing Ca(2+) per se did not induce any contraction in the absence of bradykinin. Pretreatment with either 10(-5) M 1-(beta-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenethyl)-1H imidazole hydrochloride (SKF96365) or 0.2 mM Ni(2+) abolished the contraction induced by bradykinin in the presence of extracellular Ca(2+). 4. Treatment with 10(-5) M indomethacin completely inhibited the contractile response induced by Ca(2+) replenishment, regardless of the timing of its application, before or after the application of bradykinin. 5. In endothelial cells in the valvular strips, bradykinin caused a transient [Ca(2+)](i) elevation in the presence of 1.25 mM extracellular Ca(2+), but [Ca(2+)](i) returned to the resting level within 10 min. Neither 10(-5) M SKF96365 nor 0.2 mM Ni(2+) had any effect on the peak [Ca(2+)](i) elevation, but decreased [Ca(2+)](i) in the declining phase. In the absence of extracellular Ca(2+), bradykinin induced a transient [Ca(2+)](i) elevation to a level similar to that seen in the presence of 1.25 mM extracellular Ca(2+). However, [Ca(2+)](i) then rapidly returned to the prestimulation level within 5 min. Subsequent Ca(2+) replenishment to 0.75 mM and higher in the presence of bradykinin elevated [Ca(2+)](i) to significantly higher levels than the resting level seen in the media containing 1.25 mM Ca(2+). 6. In conclusion, Ca(2+) influx in the endothelial cells is essential for bradykinin to induce endothelium-dependent contraction in the porcine interlobar renal artery. PMID- 11483702 TI - An ATP-sensitive K(+) conductance in dissociated neurones from adult rat intracardiac ganglia. AB - 1. An ATP-sensitive K(+) (K(ATP)) conductance has been identified using the perforated patch recording configuration in a population (52%) of dissociated neurones from adult rat intracardiac ganglia. The presence of the sulphonylurea receptor in approximately half of the intracardiac neurones was confirmed by labelling with fluorescent glibenclamide-BODIPY. 2. Under current clamp conditions in physiological solutions, levcromakalim (10 microM) evoked a hyperpolarization, which was inhibited by the sulphonylurea drugs glibenclamide and tolbutamide. 3. Under voltage clamp conditions in symmetrical (140 mM) K(+) solutions, bath application of levcromakalim evoked an inward current with a density of 8 pA pF(-1) at -50 mV and a slope conductance of approximately 9 nS, which reversed close to the potassium equilibrium potential (E(K)). Cell dialysis with an ATP-free intracellular solution also evoked an inward current, which was inhibited by tolbutamide. 4. Bath application of either glibenclamide (10 microM) or tolbutamide (100 microM) depolarized adult intracardiac neurones by 3-5 mV, suggesting that a K(ATP) conductance is activated under resting conditions and contributes to the resting membrane potential. 5. Activation of a membrane current by levcromakalim was concentration dependent with an EC(50) of 1.6 microM. Inhibition of the levcromakalim-activated current by glibenclamide was also concentration dependent with an IC(50) of 55 nM. 6. Metabolic inhibition with 2,4-dinitrophenol and iodoacetic acid or superfusion with hypoxic solution (P(O2) approximately 16 mmHg) also activated a membrane current. These currents exhibited similar I-V characteristics to the levcromakalim-induced current and were inhibited by glibenclamide. 7. Activation of K(ATP) channels in mammalian intracardiac neurones may contribute to changes in neural regulation of the mature heart and cardiac function during ischaemia-reperfusion. PMID- 11483703 TI - Rate dependency of delayed rectifier currents during the guinea-pig ventricular action potential. AB - 1. The action potential clamp technique was exploited to evaluate the rate dependency of delayed rectifier currents (I(Kr) and I(Ks)) during physiological electrical activity. I(Kr) and I(Ks) were measured in guinea-pig ventricular myocytes at pacing cycle lengths (CL) of 1000 and 250 ms. 2. A shorter CL, with the attendant changes in action potential shape, was associated with earlier activation and increased magnitude of both I(Kr) and I(Ks). Nonetheless, the relative contributions of I(Kr) and I(Ks) to total transmembrane current were independent of CL. 3. Shortening of diastolic interval only (constant action potential shape) enhanced I(Ks), but not I(Kr). 4. I(Kr) was increased by a change in the action potential shape only (constant diastolic interval). 5. In ramp clamp experiments, I(Kr) amplitude was directly proportional to repolarization rate at values within the low physiological range (< 1.0 V s(-1)); at higher repolarization rates proportionality became shallower and finally reversed. 6. When action potential duration (APD) was modulated by constant current injection (I-clamp), repolarization rates > 1.0 V s(-1) were associated with a reduced effect of I(Kr) block on APD. The effect of changes in repolarization rate was independent of CL and occurred in the presence of I(Ks) blockade. 7. In spite of its complexity, the behaviour of I(Kr) was accurately predicted by a numerical model based entirely on known kinetic properties of the current. 8. Both I(Kr) and I(Ks) may be increased at fast heart rates, but this may occur through completely different mechanisms. The mechanisms identified are such as to contribute to abnormal rate dependency of repolarization in prolonged repolarization syndromes. PMID- 11483704 TI - Modulation of a presynaptic hyperpolarization-activated cationic current (I(h)) at an excitatory synaptic terminal in the rat auditory brainstem. AB - 1. A hyperpolarization-activated non-specific cation current, I(h), was examined in bushy cell bodies and their giant presynaptic terminals (calyx of Held). Whole cell patch clamp recordings were made using an in vitro brain slice preparation of the cochlear nucleus and the superior olivary complex. The aim was to characterise I(h) in identified cell bodies and synaptic terminals, to examine modulation by presynaptic cAMP and to test for modulatory effects of I(h) activation on synaptic transmission. 2. Presynaptic I(h) was activated by hyperpolarizing voltage-steps, with half-activation (V(1/2)) at -94 mV. Activation time constants were voltage dependent, showing an e-fold acceleration for hyperpolarizations of -32 mV (time constant of 78 ms at -130 mV). The reversal potential of I(h) was -29 mV. It was blocked by external perfusion of 1 mM CsCl but was unaffected by BaCl(2). 3. Application of internal cAMP shifted the activation curve to more positive potentials, giving a V(1/2) of -74 mV; hence around half of the current was activated at resting membrane potentials. This shift in half-activation was mimicked by external perfusion of a membrane permeant analogue, 8-bromo-cAMP. 4. The bushy cell body I(h) showed similar properties to those of the synaptic terminal; V(1/2) was -94 mV and the reversal potential was -33 mV. Somatic I(h) was blocked by CsCl (1 mM) and was partially sensitive to BaCl(2). Somatic I(h) current density increased with postnatal age from 5 to 16 days old, suggesting that I(h) is functionally relevant during maturation of the auditory pathway. 5. The function of I(h) in regulating presynaptic excitability is subtle. I(h) had little influence on EPSC amplitude at the calyx of Held, but may be associated with propagation of the action potential at branch points. Presynaptic I(h) shares properties with both HCN1 and HCN2 recombinant channel subunits, in that it gates relatively rapidly and is modulated by internal cAMP. PMID- 11483705 TI - Drastic reduction of the slow gate of human muscle chloride channel (ClC-1) by mutation C277S. AB - 1. Single channel measurements suggest that the human muscle chloride channel ClC 1 presumably has a double barrelled structure, with a fast single protopore gate and a slow common pore gate similar to that of ClC-0, the chloride channel from Torpedo. The single point mutation C212S has been shown to abolish the slow gating of ClC-0 locking the slow gate in the open state. In order to test the hypothesis that the slow gating process found in ClC-1 corresponds to the well characterised slow gate found in ClC-0 we investigated the gating effects in ClC 1 of the homologous mutation corresponding to C212S, C277S. 2. We found that the mutation C277S strongly reduced the slow component of macroscopic gating relaxations at negative and at positive voltages. 3. Time constants of the fast gating relaxations were not affected by the mutation but the minimal open probability of the fast gate at negative voltages was slightly reduced to 0.08 compared with the WT value of 0.22. 4. Additionally, we characterised the block of WT ClC-1 and mutant C277S by the S(-) enantiomer of CPB (2-(p-chlorophenoxy) butyric acid), and found that the block is practically unaffected by the mutation suggesting that CPB does not interact with the slow gate of ClC-1. 5. We conclude that the slow and fast gating processes of ClC-1, respectively, reflect the slow common pore gate and the single protopore gate of the double-barrelled ClC-1 channel. PMID- 11483706 TI - Measurement of chloride flux associated with the myogenic response in rat cerebral arteries. AB - 1. Self-referencing ion-selective (SERIS) electrodes were used to measure the temperature and pressure dependence of Cl(-) efflux, during myogenic contraction of pressurized rat cerebral resistance arteries. 2. At room temperature (18-21 degrees C), a small, pressure-independent Cl(-) efflux was measured. On warming to 37 degrees C, arteries developed pressure-dependent myogenic tone, and this was associated with a pressure-dependent increase in Cl(-) efflux (n = 5). 3. Both myogenic tone and the pressure- and temperature-dependent Cl(-) efflux were abolished on application of 10 microM tamoxifen, a Cl(-) channel blocker (IC(50) 3.75 +/- 0.2 microM). Tamoxifen (10 microM) also prevented contraction to 60 mM K(+), suggesting non-specific effects of tamoxifen (n = 5). 4. Myogenic tone was abolished by 2 microM nimodipine, but Cl(-) efflux was unaffected. In the presence of nimodipine, 10 microM tamoxifen still abolished pressure- and temperature-dependent Cl(-) efflux (n = 3). 5. In summary, a Cl(-) efflux can be measured from rat cerebral arteries, with a temperature dependence that is closely correlated with myogenic contraction. We conclude that Cl(-) efflux through Cl(-) channels contributes to the depolarization associated with myogenic contraction. PMID- 11483707 TI - Developmental expression of the novel voltage-gated sodium channel auxiliary subunit beta3, in rat CNS. AB - 1. We have compared the mRNA distribution of sodium channel alpha subunits known to be expressed during development with the known auxiliary subunits Nabeta1.1 and Nabeta2.1 and the novel, recently cloned subunit, beta3. 2. In situ hybridisation studies demonstrated high levels of Nav1.2, Nav1.3, Nav1.6 and beta3 mRNA at embryonic stages whilst Nabeta1.1 and Nabeta2.1 mRNA was absent throughout this period. 3. Nabeta1.1 and Nabeta2.1 expression occurred after postnatal day 3 (P3), increasing steadily in most brain regions until adulthood. beta3 expression differentially decreased after P3 in certain areas but remained high in the hippocampus and striatum. 4. Emulsion-dipped slides showed co localisation of beta3 with Nav1.3 mRNA in areas of the CNS suggesting that these subunits may be capable of functional interaction. 5. Co-expression in Xenopus oocytes revealed that beta3 could modify the properties of Nav1.3; beta3 changed the equilibrium of Nav1.3 between the fast and slow gating modes and caused a negative shift in the voltage dependence of activation and inactivation. 6. In conclusion, beta3 is shown to be the predominant beta subunit expressed during development and is capable of modulating the kinetic properties of the embryonic Nav1.3 subunit. These findings provide new information regarding the nature and properties of voltage-gated sodium channels during development. PMID- 11483708 TI - State-dependent action of grayanotoxin I on Na(+) channels in frog ventricular myocytes. AB - 1. Distinct properties of grayanotoxin (GTX) among other lipid-soluble toxins were elucidated by quantitative analysis made on the Na(+) channel in frog ventricular myocytes. 2. GTX-modified current (I(GTX)) was induced strictly in proportion to the open probability of Na(+) channels during preconditioning pulses irrespective of its duration, amplitude or partial removal of inactivation by chloramine-T. This confirms that GTX binds to the Na(+) channel exclusively in its open state, while batrachotoxin (BTX) was reported to be capable of modifying slow-inactivated Na(+) channels, and veratridine exhibited voltage-dependent modification. 3. The GTX-modified channel did not show any inactivation property, which is different from reported results with veratridine and BTX. 4. Estimated unbinding rates of GTX were in reverse proportion to the activation curve of GTX modified Na(+) channels. This was not the previously reported case with veratridine. 5. A model including unbinding kinetics of GTX and slow inactivation of unmodified Na(+) channels in which GTX was permitted to bind only to the open state of Na(+) channels indicated that unbinding reactions of GTX occur only in the closed state. PMID- 11483709 TI - Histaminergic modulation of GABAergic transmission in rat ventromedial hypothalamic neurones. AB - 1. The ventromedial nucleus of the hypothalamus (VMH) is a key nucleus in the homeostatic regulation of neuroendocrine and behavioural functions. In mechanically dissociated rat VMH neurones with attached native presynaptic nerve endings, GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) were recorded using the nystatin perforated patch recording mode under voltage-clamp conditions. 2. Histamine reversibly inhibited the sIPSC frequency in a concentration-dependent manner without affecting the mean current amplitude. The selective histamine receptor type 3 (H(3)) agonist imetit (100 nM) mimicked this effect and it was completely abolished by the selective H(3) receptor antagonists clobenpropit (3 microM) and thioperamide (10 microM). 3. The GTP-binding protein inhibitor N-ethylmaleimide (10 microM) removed the histaminergic inhibition of GABAergic sIPSCs. 4. Elimination of external Ca(2+) reduced the GABAergic sIPSC frequency without affecting the distribution of current amplitudes. In this condition, the inhibitory effect of imetit on the sIPSC frequency completely disappeared, suggesting that the histaminergic inhibition requires extracellular Ca(2+). 5. The P/Q-type Ca(2+) channel blocker omega-agatoxin IVA (300 nM) attenuated the histaminergic inhibition of the GABAergic sIPSC frequency, but neither the N-type Ca(2+) channel blocker omega-conotoxin GVIA (3 microM) nor the L-type Ca(2+) channel blocker nicardipine (3 microM) was effective. 6. Activation of adenylyl cyclase with forskolin (10 microM) had no effect on histaminergic inhibition of the sIPSCs. 7. In conclusion, histamine inhibits spontaneous GABA release from presynaptic nerve terminals projecting to VMH neurones by inhibiting presynaptic P/Q-type Ca(2+) channels via a G-protein coupled to H(3) receptors and this may modulate the excitability of VMH neurones. PMID- 11483710 TI - Cannabinoid actions on rat superficial medullary dorsal horn neurons in vitro. AB - 1. This study examined the cellular actions of cannabinoids on neurons in the substantia gelatinosa of the spinal trigeminal nucleus pars caudalis, using whole cell and perforated patch recording in brain slices. 2. The cannabinoid agonist WIN55,212-2 (3 microM) decreased the amplitude of both GABAergic and glycinergic electrically evoked inhibitory postsynaptic currents (IPSCs) by 35 and 41 %, respectively. This inhibition was completely reversed by the CB(1) receptor selective antagonist N-piperidino-5-(4-chlorophenyl)-l-(2,4-dichlorophenyl)-4 methyl-3-pyrazole-carboxamide) (SR141716A, 3 microM). WIN55,212-2 also produced relative facilitation of the second evoked IPSC to paired stimuli. 3. WIN55,212-2 decreased the rate of both GABAergic and glycinergic miniature IPSCs by 44 and 34 %, respectively, without changing their amplitude distributions or kinetics. 4. WIN55,212-2 did not affect the amplitude of electrically evoked non-NMDA glutamatergic excitatory postsynaptic currents (EPSCs). 5. WIN55,212-2 produced no postsynaptic membrane current and had no significant effect on membrane conductance over a range of membrane potentials (-60 to -130 mV). 6. These results suggest that, within the superficial medullary dorsal horn, cannabinoids presynaptically inhibit GABAergic and glycinergic neurotransmission. At the cellular level, the analgesic action of cannabinoids on these medullary dorsal horn neurons therefore differs from that of mu-opioids, which have both pre- and postsynaptic actions. PMID- 11483711 TI - Protein kinase C activation potentiates gating of the vanilloid receptor VR1 by capsaicin, protons, heat and anandamide. AB - 1. The effects of activation of protein kinase C (PKC) on membrane currents gated by capsaicin, protons, heat and anandamide were investigated in primary sensory neurones from neonatal rat dorsal root ganglia (DRG) and in HEK293 cells (human embryonic kidney cell line) transiently or stably expressing the human vanilloid receptor hVR1. 2. Maximal activation of PKC by a brief application of phorbol 12 myristate 13-acetate (PMA) increased the mean membrane current activated by a low concentration of capsaicin by 1.65-fold in DRG neurones and 2.18-fold in stably transfected HEK293 cells. Bradykinin, which activates PKC, also enhanced the response to capsaicin in DRG neurones. The specific PKC inhibitor RO31-8220 prevented the enhancement caused by PMA. 3. Activation of PKC did not enhance the membrane current at high concentrations of capsaicin, showing that PKC activation increases the probability of channel opening rather than unmasking channels. 4. Application of PMA alone activated an inward current in HEK293 cells transiently transfected with VR1. The current was suppressed by the VR1 antagonist capsazepine. PMA did not, however, activate a current in the large majority of DRG neurones nor in HEK293 cells stably transfected with VR1. 5. Removing external Ca(2+) enhanced the response to a low concentration of capsaicin 2.40 fold in DRG neurones and 3.42-fold in HEK293 cells. Activation of PKC in zero Ca(2+) produced no further enhancement of the response to capsaicin in either DRG neurones or HEK293 cells stably transfected with VR1. 6. The effects of PKC activation on the membrane current gated by heat, anandamide and low pH were qualitatively similar to those on the capsaicin-gated current. 7. The absence of a current activated by PMA in most DRG neurones or in stably transfected HEK293 cells suggests that activation of PKC does not directly open VR1 channels, but instead increases the probability that they will be activated by capsaicin, heat, low pH or anandamide. Removal of calcium also potentiates activation, and PKC activation then has no further effect. The results are consistent with a model in which phosphorylation of VR1 by PKC increases the probability of channel gating by agonists, and in which dephosphorylation occurs by a calcium-dependent process. PMID- 11483712 TI - Functional coupling between nitric oxide synthesis and VIP release within enteric nerve terminals of the rat: involvement of protein kinase G and phosphodiesterase 5. AB - 1. The subcellular mechanisms involved in the effect of nitric oxide (NO) on the release of vasoactive intestinal polypeptide (VIP) were examined in synaptosomes isolated from rat small intestine. 2. VIP release was stimulated by the NO donor SNAP (10(-7)-10(-4) M) in an oxyhaemoglobin-sensitive manner. The presence of the guanylate cyclase inhibitor ODQ (10(-5) M), or inhibition of protein kinase G (PKG) by KT 5823 (3 x 10(-6) M) or Rp-8Br-PET-cGMPS (5 x 10(-7) M), antagonized the SNAP-induced VIP release, suggesting a regulatory role of PKG, confirming previously published data from enteric ganglia. This finding was further supported by the fact that direct PKG activation by the stable cGMP analogue 8 pCPT-cGMP stimulated VIP secretion to the same extent as SNAP. 3. Basal VIP secretion was enhanced in the presence of zaprinast, an inhibitor of cGMP dependent phosphodiesterase 5 (PDE 5), suggesting a functional role of PDE 5 in NO-cGMP signalling. Supportive evidence for this finding was obtained by demonstration of the presence of PDE 5 using RT-PCR. 4. Stimulation of endogenous NO production by L-arginine was also effective in releasing VIP. The effect was abolished in the presence of KT 5823, but was insensitive to oxyhaemoglobin (10( 3) M), suggesting that an interaction between NO and VIP is likely to occur within the same nerve terminal rather than between terminals. 5. NO synthesis was not affected by VIP (10(-8)-10(-5) M), suggesting that there is no feedback regulation between the NO and the VIP pathways. 6. These findings support the notion that an anatomical and functional interrelationship exists between NO and VIP in enteric nerve terminals and that complex signalling mechanisms involving PKG and PDE 5 contribute to NO-induced VIP release. PMID- 11483713 TI - Differential expression and regulation of AE2 anion exchanger subtypes in rabbit parietal and mucous cells. AB - 1. The anion exchanger isoform 2 (AE2) gene encodes three subtypes (AE2a, b and c), which have different N-termini and tissue distributions. AE2 is expressed at high levels in the stomach, where it is thought to mediate basolateral base exit during acid production. The present study investigated if the three AE2 subtypes are differentially expressed and regulated in different cell types within the gastric mucosa. 2. The cloning strategy to obtain rabbit AE2a, b and c cDNAs combined genomic PCR and RT-PCR based on primers deduced from the rat sequences. Semiquantitative RT-PCR using homologous primers revealed much higher AE2 mRNA expression in rabbit parietal cells (PCs) than in mucous cells (MCs). The subtype expression pattern was AE2b >> AE2c > or = AE2a in PCs and AE2a >AE2b >> AE2c in MCs. Sequence analysis revealed the presence of a highly conserved protein kinase C (PKC) consensus sequence in the AE2a alternative N-terminus. 3. Maximal Cl(-) HCO(3)(-) exchange rates, measured fluorometrically in BCECF-loaded cultured gastric cells, were much higher in PCs than MCs. PKC activation by phorbol ester stimulated maximal Cl(-)-HCO(3)(-) exchange rates in MCs but not in PCs, whereas forskolin had no effect in each cell type. 4. In summary, rabbit PCs and MCs, which originate from the same gastric stem cell population, display a completely different AE2 subtype expression pattern. Therefore, AE2 subtype expression is not organ specific but cell type specific. The different regulation of anion exchange in parietal and mucous cells suggests that AE2 subtypes may be differentially regulated. PMID- 11483714 TI - Actions of nociceptin/orphanin FQ and other prepronociceptin products on rat rostral ventromedial medulla neurons in vitro. AB - 1. Whole-cell patch clamp recordings were made from rat rostral ventromedial medulla (RVM) neurons in vitro to investigate the cellular actions of the opioid like receptor ORL1 (NOP), ligand nociceptin/orphanin FQ and other putative prepronociceptin products. 2. Primary and secondary RVM neurons were identified as responding to the kappa-opioid receptor agonist U-69593 (300 nM to 1 microM) and the mu- and delta-opioid receptor agonist met-enkephalin (10 microM), respectively. Both primary and secondary RVM neurons responded to nociceptin (3 nM to 1 microM) with an outward current that reversed polarity at -115 mV in brain slices and with inhibition of Ca(2+) channel currents in acutely isolated cells. 3. The putative ORL1 antagonist J-113397 (1 microM) produced no change in membrane current and abolished the outward current produced by nociceptin (100 nM). In contrast, Phe(1)psi(CH(2)-NH)Gly(2)]-nociceptin-(1-13)NH(2) (300 nM to 1 microM) alone produced an outward current and partially reduced the outward current produced by nociceptin (300 nM) when co-applied. 4. In brain slices nociceptin (300 nM) reduced the amplitude of evoked GABA(A) receptor-mediated inhibitory postsynaptic currents (IPSCs) but not non-NMDA receptor-mediated excitatory postsynaptic currents (EPSCs). 5. Met-enkephalin (10 microM), but not nociceptin (300 nM), reduced the rate of spontaneous miniature IPSCs in normal external potassium solution (K(+) 2.5 mM). In high external potassium (K(+) 17.5 mM), nociceptin reduced the rate of miniature IPSCs in the presence (Ca(2+) 2.4 mM, Mg(2+) 1.2 mM) but not in the absence of external calcium (Ca(2+) 0 mM, Mg(2+) 10 mM, Cd(2+) 10 microM). Nociceptin and met-enkephalin had no effect on the amplitude of miniature IPSCs. 6. The putative nociceptin precursor products nocistatin (rat prepronociceptin(125-132)) and rat prepronociceptin(154-181) had no effect on membrane currents, evoked IPSCs and evoked EPSCs. 7. These results indicate that nociceptin acts via the ORL1 receptor to directly inhibit both primary and secondary RVM neurons by activating a potassium conductance and by inhibiting calcium conductances. In addition, nociceptin inhibits GABA release within the RVM via a presynaptic Ca(2+)-dependent mechanism. Thus, nociceptin has the potential to exert both disinhibitory and inhibitory effects on neuronal action potential firing within the RVM. PMID- 11483715 TI - Developmental regulation of transmitter release at the calyx of Held in rat auditory brainstem. AB - 1. Postnatal development of synaptic efficacy was studied at a single glutamatergic synapse formed by the calyx of Held in the rat brainstem. Throughout postnatal development from day 7 (P7) to day 14 (P14), both the excitatory postsynaptic currents (EPSCs) evoked at a basal frequency (0.05 Hz) and spontaneous miniature EPSCs (mEPSCs) remained similar in their mean amplitudes, but became faster in their decay times. 2. During repetitive stimulation at 1-100 Hz, EPSCs underwent a depression. The magnitude of the depression significantly decreased from P7 to P14, whereas the time course of recovery from depression (after 10 Hz stimulation) remained similar throughout development. 3. The size of the readily releasable pool (RRP) of synaptic vesicles (N) and the release probability (p) were estimated from the cumulative amplitude histogram of EPSCs during high frequency stimulation. From P7 to P14, N increased 2-fold, whereas p decreased to a similar extent. 4. The open channel blocker MK-801 caused an activity-dependent attenuation of NMDA receptor-mediated EPSCs. The blocking rate became slower from P7 to P14, further supporting the developmental decrease in p. 5. Given that the mean amplitudes of mEPSCs (q) and evoked EPSCs (Npq) remain constant throughout the developmental period, these results suggest that a developmental increase in N compensates for a concomitant decrease in p. We conclude that the developmental decrease in the release probability will establish a stable synapse at which only a small fraction of releasable synaptic vesicles is depleted during high frequency transmission. PMID- 11483716 TI - Contribution of spindle reflexes to post-inspiratory activity in the canine external intercostal muscles. AB - 1. The external intercostal muscles have greater post-inspiratory activity than the parasternal intercostal muscles and are more abundantly supplied with muscle spindles. In the present study, the hypothesis was tested that spindle afferent inputs play a major role in determining this activity. 2. The electrical activity of the external and parasternal intercostal muscles in the rostral interspaces was recorded in anaesthetized spontaneously breathing dogs, and the ribs were manipulated so as to alter their normal caudal displacement and the normal lengthening of the muscles in early expiration. 3. Post-inspiratory activity in the external intercostal muscles showed a reflex decrease when the caudal motion of the ribs and the lengthening of the muscles was impeded, and it showed a reflex increase when the rate of caudal rib motion and muscle lengthening was increased. In contrast, the small post-inspiratory activity in the parasternal intercostal muscles remained unchanged. 4. When the two ribs making up the interspace investigated were locked to keep muscle length constant, post inspiratory activity in the external intercostal muscles was reduced and no longer responded to cranial rib manipulation. 5. These observations confirm that afferent inputs from muscle receptors, presumably muscle spindles, are a primary determinant of post-inspiratory activity in the canine external intercostal muscles. In anaesthetized animals, the contribution of central control mechanisms to this activity is small. PMID- 11483717 TI - The ventilatory response to arousal from sleep is not fully explained by differences in CO(2) levels between sleep and wakefulness. AB - 1. Arousal from sleep is associated with transient stimulation of ventilation above normal waking levels that predisposes to subsequent breathing instability and central apnoea. The transient hyperpnoea at arousal is normally explained by differences in arterial partial pressure of CO(2) (P(a,CO2)) between sleep and wakefulness, with a higher P(a,CO2) in sleep leading to stimulation of ventilation at arousal according to the awake ventilatory response to CO(2). Surprisingly, however, the validity of this current model in fully explaining the increased ventilation at arousal from sleep has not been directly tested. 2. This study tests the hypothesis that the level of ventilation at arousal from non rapid eye movement (non-REM) sleep is greater than that produced by elevating P(a,CO2) in wakefulness to the sleeping level, i.e. the ventilation predicted by the current model. 3. Studies were performed in five dogs. Inspired CO(2) was used to increase end-tidal partial pressure of CO(2) (P(ET,CO2)) in wakefulness and measure the ventilatory response. The same P(ET,CO2) was then maintained in non-REM sleep. Ventilation was measured for 10 breaths before and after arousal from non-REM sleep induced by a 72 dB tone. 4. Arousal from sleep produced a transient surge in ventilation of 1.42 +/- 0.35 l min(-1) (P = 0.005). This increased ventilation was due to arousal from sleep per se as the tone alone produced no change in awake ventilation. In support of the hypothesis, ventilation at wake onset from sleep was greater by 0.83 +/- 0.28 l min(-1) (P = 0.031) than the ventilation elicited in wakefulness by raising P(ET,CO2) to the sleeping level. 5. The results show that > 50 % of the increase in ventilation at wake onset from sleep is not attributable to the awake ventilatory response to the elevated P(a,CO2) that was previously present in sleep. This result leads to important modifications of the physiological model currently used to explain the ventilatory consequences of arousal from sleep. PMID- 11483718 TI - Changes in intracortical excitability induced by stimulation of wrist afferents in man. AB - 1. Inhibitory and facilitatory neuronal circuits may be explored in the human motor cortex by double pulse transcranial magnetic stimulation (TMS). At short interstimulus intervals (2-5 ms), conditioned motor-evoked potentials (MEPs) are reduced (intracortical inhibition, ICI), whereas they are facilitated at longer interstimulus intervals (8-25 ms; intracortical facilitation, ICF). The aim of this study was to investigate the effects of homonymous and antagonist nerve stimulation on the intracortical inhibition and facilitation in the cortical areas that control the wrist extensor and flexor radialis muscles. 2. Sixteen subjects were asked to contract either their wrist extensor or flexor muscles. The MEP evoked by a test TMS (at 1.2 x MEP threshold) and recorded in the target muscle was then conditioned by subthreshold TMS (at 0.8 x MEP threshold) 2 and 14 ms before the test TMS. The median and radial nerves were stimulated at 0.8 x motor threshold (MT). 3. In both flexor and extensor muscles, antagonist nerve stimulation 40 ms before the test TMS decreased ICI and increased ICF. In contrast, homonymous nerve stimulation had no effect on ICI and ICF. 4. The intensity of the antagonist nerve stimulation required to alter ICI and ICF was as low as 0.6 x MT, which suggests that thick diameter afferents may be involved. The nerve stimulation had to be applied 35-45 ms prior to the test TMS to alter significantly the intracortical excitability. 5. Cutaneous afferents were probably not responsible for the alterations of intracortical excitability, since cutaneous stimulation had no effect on either ICI or ICF at the investigated intervals. 6. The present data suggest that antagonist muscular afferent inputs may evoke reciprocal facilitation or disinhibition at the cortical level. This pattern of antagonist sensory afferent effects may be of significance for control of the wrist extensor and flexor muscles when used as synergists during manipulatory finger movements and gripping tasks. PMID- 11483719 TI - Motor unit behaviour and contractile changes during fatigue in the human first dorsal interosseus. AB - 1. In 67 single motor units, the mechanical properties, the recruitment and derecruitment thresholds, and the discharge rates were recorded concurrently in the first dorsal interosseus (FDI) of human subjects during intermittent fatiguing contractions. The task consisted of isometric ramp-and-hold contractions performed at 50 % of the maximal voluntary contraction (MVC). The purpose of this study was to examine the influence of fatigue on the behaviour of motor units with a wide range of activation thresholds. 2. For low-threshold (< 25 % MVC) motor units, the mean twitch force increased with fatigue and the recruitment threshold either did not change or increased. In contrast, the twitch force and the activation threshold decreased for the high-threshold (> 25 % MVC) units. The observation that in low-threshold motor units a quick stretch of the muscle at the end of the test reset the unit force and recruitment threshold to the prefatigue value suggests a significant role for fatigue-related changes in muscle stiffness but not twitch potentiation or motor unit synchronization. 3. Although the central drive intensified during the fatigue test, as indicated by an increase in surface electromyogram (EMG), the discharge rate of the motor units during the hold phase of each contraction decreased progressively over the course of the task for motor units that were recruited at the beginning of the test, especially the low-threshold units. In contrast, the discharge rates of newly activated units first increased and then decreased. 4. Such divergent behaviour of low- and high-threshold motor units could not be individually controlled by the central drive to the motoneurone pool. Rather, the different behaviours must be the consequence of variable contributions from motoneurone adaptation and afferent feedback from the muscle during the fatiguing contraction. PMID- 11483720 TI - H-reflex modulation during passive lengthening and shortening of the human triceps surae. AB - 1. The present study investigated the effects of lengthening and shortening actions on H-reflex amplitude. H-reflexes were evoked in the soleus (SOL) and medial gastrocnemius (MG) of human subjects during passive isometric, lengthening and shortening actions performed at angular velocities of 0, +/-2, +/-5 and +/-15 deg s(-1). 2. H-reflex amplitudes in both SOL and MG were significantly depressed during passive lengthening actions and facilitated during passive shortening actions, when compared with the isometric H-reflex amplitude. 3. Four experiments were performed in which the latencies from the onset of movement to delivery of the stimulus were altered. Passive H-reflex modulation during lengthening actions was found to begin at latencies of less than 60 ms suggesting that this inhibition was due to peripheral and/or spinal mechanisms. 4. It is postulated that the H-reflex modulation seen in the present study is related to the tonic discharge of muscle spindle afferents and the consequent effects of transmission within the Ia pathway. Inhibition of the H-reflex at less than 60 ms after the onset of muscle lengthening may be attributed to several mechanisms, which cannot be distinguished using the current protocol. These may include the inability to evoke volleys in Ia fibres that are refractory following muscle spindle discharge during rapid muscle lengthening, a reduced probability of transmitter release from the presynaptic terminal (homosynaptic post-activation depression) and presynaptic inhibition of Ia afferents from plantar flexor agonists. Short latency facilitation of the H-reflex may be attributed to temporal summation of excitatory postsynaptic potentials arising from muscle spindle afferents during rapid muscle lengthening. At longer latencies, presynaptic inhibition of Ia afferents cannot be excluded as a potential inhibitory mechanism. PMID- 11483721 TI - Group II muscle afferents probably contribute to the medium latency soleus stretch reflex during walking in humans. AB - 1. The objective of this study was to determine which afferents contribute to the medium latency response of the soleus stretch reflex resulting from an unexpected perturbation during human walking. 2. Fourteen healthy subjects walked on a treadmill at approximately 3.5 km h(-1) with the left ankle attached to a portable stretching device. The soleus stretch reflex was elicited by applying small amplitude (approximately 8 deg) dorsiflexion perturbations 200 ms after heel contact. 3. Short and medium latency responses were observed with latencies of 55 +/- 5 and 78 +/- 6 ms, respectively. The short latency response was velocity sensitive (P < 0.001), while the medium latency response was not (P = 0.725). 4. Nerve cooling increased the delay of the medium latency component to a greater extent than that of the short latency component (P < 0.005). 5. Ischaemia strongly decreased the short latency component (P = 0.004), whereas the medium latency component was unchanged (P = 0.437). 6. Two hours after the ingestion of tizanidine, an alpha(2)-adrenergic receptor agonist known to selectively depress the transmission in the group II afferent pathway, the medium latency reflex was strongly depressed (P = 0.007), whereas the short latency component was unchanged (P = 0.653). 7. An ankle block with lidocaine hydrochloride was performed to suppress the cutaneous afferents of the foot and ankle. Neither the short (P = 0.453) nor medium (P = 0.310) latency reflexes were changed. 8. Our results support the hypothesis that, during walking the medium latency component of the stretch reflex resulting from an unexpected perturbation is contributed to by group II muscle afferents. PMID- 11483722 TI - Genetic analysis of the pestivirus nonstructural coding region: defects in the NS5A unit can be complemented in trans. AB - The functional analysis of molecular determinants which control the replication of pestiviruses was considerably facilitated by the finding that subgenomic forms of the positive-strand RNA genome of BVDV (bovine viral diarrhea virus) are capable of autonomous replication in transfected host cells. The prototype replicon, BVDV DI9c, consists of the genomic 5' and 3' untranslated regions and a truncated open reading frame (ORF) encoding mainly the nonstructural proteins NS3, NS4A, NS4B, NS5A, and NS5B. To gain insight into which of these proteins are essential for viral replication and whether they act in cis or in trans, we introduced a large spectrum of in-frame mutations into the DI9c ORF. Tests of the mutant RNAs in terms of their replication capacity and their ability to support translation and cleavage of the nonstructural polyprotein, and whether defects could be rescued in trans, yielded the following results. (i) RNA replication was found to be dependent on the expression of each of the DI9c-encoded mature proteins NS3 to NS5B (and the known associated enzymatic activities). In the same context, a finely balanced molar ratio of the diverse proteolytic processing products was indicated to be crucial for the formation of an active catalytic replication complex. (ii) Synthesis of negative-strand intermediate and progeny positive-strand RNA was observed to be strictly coupled with all functional DI9c ORF derivatives. NS3 to NS5B were hence suggested to play a pivotal role even during early steps of the viral replication pathway. (iii) Mutations in the NS3 and NS4B units which generated nonfunctional or less functional RNAs were determined to be cis dominant. Likewise, lethal alterations in the NS4A and NS5B regions were invariably noncomplementable. (iv) In surprising contrast, replication of functional and nonfunctional NS5A mutants could be clearly enhanced and restored, respectively. In summary, our data provide initial insights into the organization of the pestivirus replication machinery. PMID- 11483723 TI - Different levels of T-cell receptor triggering induce distinct functions in hepatitis B and hepatitis C virus-specific human CD4(+) T-cell clones. AB - CD4(+) T cells play a major role in the host defense against viruses and intracellular microbes. During the natural course of such an infection, specific CD4(+) T cells are exposed to a wide range of antigen concentrations depending on the body compartment and the stage of disease. While epitope variants trigger only subsets of T-cell effector functions, the response of virus-specific CD4(+) T cells to various concentrations of the wild-type antigen has not been systematically studied. We stimulated hepatitis B virus core- and hepatitis C virus NS3-specific CD4(+) T-cell clones which had been isolated from patients with acute hepatitis during viral clearance with a wide range of specific antigen concentrations and determined the phenotypic changes and the induction of T-cell effector functions in relation to T-cell receptor internalization. A low antigen concentration induced the expression of T-cell activation markers and adhesion molecules in CD4(+) T-cell clones in the absence of cytokine secretion and proliferation. The expression of CD25, HLA-DR, CD69, and intercellular cell adhesion molecule 1 increased as soon as T-cell receptor internalization became detectable. A 30- to 100-fold-higher antigen concentration, corresponding to the internalization of 20 to 30% of T-cell receptor molecules, however, was required for the induction of proliferation as well as for gamma interferon and interleukin-4 secretion. These data indicate that virus-specific CD4(+) T cells can respond to specific antigen in a graded manner depending on the antigen concentration, which may have implications for a coordinate regulation of specific CD4(+) T-cell responses. PMID- 11483724 TI - The leader protein of Theiler's virus inhibits immediate-early alpha/beta interferon production. AB - Theiler's virus is a picornavirus responsible for a persistent infection of the central nervous system of the mouse, leading to a chronic demyelinating disease considered to be a model for multiple sclerosis. The leader (L) protein encoded by Theiler's virus is a 76-amino-acid-long peptide containing a zinc-binding motif. This motif is conserved in the L proteins of all cardioviruses, including encephalomyocarditis virus. The L protein of Theiler's virus was suggested to interfere with the alpha/beta interferon (IFN-alpha/beta) response (W.-P. Kong, G. D. Ghadge, and R. P. Roos, Proc. Natl. Acad. Sci. USA 91:1796-1800, 1994). We show that expression of the L protein indeed inhibits the production of alpha/beta interferon by infected L929 cells. The L protein specifically inhibits the transcription of the IFN-alpha4 and IFN-beta genes, which are known to be activated early in response to viral infection. Mutation of the zinc finger was sufficient to block the anti-interferon activity, outlining the importance of this motif in the L protein function. In agreement with the anti-interferon role of the L protein, a virus bearing a mutation in the zinc-binding motif was dramatically impaired in its ability to persist in the central nervous system of SJL/J mice. PMID- 11483725 TI - Identification of a putative coreceptor on Vero cells that participates in dengue 4 virus infection. AB - Dengue virus infects target cells by attaching to a cell surface receptor through the envelope (E) glycoprotein, located on the surface of the viral membrane. On Vero and BHK cells, heparan sulfate (HS) moieties of proteoglycans are the receptors for dengue virus; however, additional proteins have also been described as putative dengue virus receptors on C6/36, HL60, and BM cells. HS can also act as a receptor for other types of viruses or as an attachment molecule for viruses that require additional host cell molecules to allow viral penetration. In this study we searched for molecules other than HS that could participate in dengue virus infection of Vero cells. Labeled dengue 4 virus bound with high affinity to two molecules of 74 and 44 kDa. Binding of dengue virus to the 74-kDa molecule was susceptible to protease and sodium periodate treatment and resistant to heparinase treatments. Lectins such as concanavalin A and wheat germ agglutinin prevented dengue virus binding to both the 74- and the 44-kDa protein in overlay assays, while phytohemagglutinin P did not affect binding, suggesting that carbohydrate residues (alpha-mannose or N-acetylglucosamine) are important in virus binding to host cells. Protease susceptibility, biotin labeling, and immunofluorescence with a polyclonal antibody raised against the 74-kDa protein consistently identified the protein on the surfaces of Vero cells. Moreover, the antibody against the 74-kDa protein was able to inhibit dengue virus infection. These data suggest that HS might serve as a primary receptor, probably concentrating virus particles on the surfaces of Vero cells, and then other molecules, such as the 74-kDa protein, might participate as coreceptors in viral penetration. The 74-kDa protein possibly constitutes part of a putative receptor complex for dengue virus infection of Vero cells. PMID- 11483726 TI - Salicylates inhibit flavivirus replication independently of blocking nuclear factor kappa B activation. AB - Flaviviruses comprise a positive-sense RNA genome that replicates exclusively in the cytoplasm of infected cells. Whether flaviviruses require an activated nuclear factor(s) to complete their life cycle and trigger apoptosis in infected cells remains elusive. Flavivirus infections quickly activate nuclear factor kappa B (NF-kappaB), and salicylates have been shown to inhibit NF-kappaB activation. In this study, we investigated whether salicylates suppress flavivirus replication and virus-induced apoptosis in cultured cells. In a dose dependent inhibition, we found salicylates within a range of 1 to 5 mM not only restricted flavivirus replication but also abrogated flavivirus-triggered apoptosis. However, flavivirus replication was not affected by a specific NF kappaB peptide inhibitor, SN50, and a proteosome inhibitor, lactacystin. Flaviviruses also replicated and triggered apoptosis in cells stably expressing IkappaBalpha-DeltaN, a dominant-negative mutant that antagonizes NF-kappaB activation, as readily as in wild-type BHK-21 cells, suggesting that NF-kappaB activation is not essential for either flavivirus replication or flavivirus induced apoptosis. Salicylates still diminished flavivirus replication and blocked apoptosis in the same IkappaBalpha-DeltaN cells. This inhibition of flaviviruses by salicylates could be partially reversed by a specific p38 mitogen activated protein (MAP) kinase inhibitor, SB203580. Together, these results show that the mechanism by which salicylates suppress flavivirus infection may involve p38 MAP kinase activity but is independent of blocking the NF-kappaB pathway. PMID- 11483727 TI - Ex vivo stimulation and expansion of both CD4(+) and CD8(+) T cells from peripheral blood mononuclear cells of human cytomegalovirus-seropositive blood donors by using a soluble recombinant chimeric protein, IE1-pp65. AB - The transfer of anti-human cytomegalovirus (HCMV) effector T cells to allogeneic bone marrow recipients results in protection from HCMV disease associated with transplantation, suggesting the direct control of CMV replication by T cells. IE1 and pp65 proteins, both targets of CD4(+) and CD8(+) T cells, are considered the best candidates for immunotherapy and vaccine design against HCMV. In this report, we describe the purification of a 165-kDa chimeric protein, IE1-pp65, and its use for in vitro stimulation and expansion of anti-HCMV CD4(+) and CD8(+) T cells from peripheral blood mononuclear cells (PBMC) of HCMV-seropositive donors. We demonstrate that an important proportion of anti-HCMV CD4(+) T cells was directed against IE1-pp65 in HCMV-seropositive donors and that the protein induced activation of HLA-DR3-restricted anti-IE1 CD4(+) T-cell clones, as assessed by gamma interferon (IFN-gamma) secretion and cytotoxicity. Moreover, soluble IE1-pp65 stimulated and expanded anti-pp65 CD8(+) T cells from PBMC of HLA-A2, HLA-B35, and HLA-B7 HCMV-seropositive blood donors, as demonstrated by cytotoxicity, intracellular IFN-gamma labeling, and quantitation of peptide specific CD8(+) cells using an HLA-A2-peptide tetramer and staining of intracellular IFN-gamma. These results suggest that soluble IE1-pp65 may provide an alternative to infectious viruses used in current adoptive strategies of immunotherapy. PMID- 11483728 TI - Immunization with a pentameric L1 fusion protein protects against papillomavirus infection. AB - The prophylactic papillomavirus vaccines currently in clinical trials are composed of viral L1 capsid protein that is synthesized in eukaryotic expression systems and purified in the form of virus-like particles (VLPs). To evaluate whether VLPs are necessary for effective vaccination, we expressed the L1 protein as a glutathione S-transferase (GST) fusion protein in Escherichia coli and assayed its immunogenic activity in an established canine oral papillomavirus (COPV) model that previously validated the efficacy of VLP vaccines. The GST-COPV L1 fusion protein formed pentamers, but these capsomere-like structures did not assemble into VLPs. Despite the lack of VLP formation, the GST-COPV L1 protein retained its native conformation as determined by reactivity with conformation specific anti-COPV antibodies. Most importantly, the GST-COPV L1 pentamers completely protected dogs from high-dose viral infection of their oral mucosa. L1 fusion proteins expressed in bacteria represent an economical alternative to VLPs as a human papillomavirus vaccine. PMID- 11483729 TI - Activity of the hepatitis A virus IRES requires association between the cap binding translation initiation factor (eIF4E) and eIF4G. AB - The question of whether translation initiation factor eIF4E and the complete eIF4G polypeptide are required for initiation dependent on the IRES (internal ribosome entry site) of hepatitis A virus (HAV) has been examined using in vitro translation in standard and eIF4G-depleted rabbit reticulocyte lysates. In agreement with previous publications, the HAV IRES is unique among all picornavirus IRESs in that it was inhibited if translation initiation factor eIF4G was cleaved by foot-and-mouth disease L-proteases. In addition, the HAV IRES was inhibited by addition of eIF4E-binding protein 1, which binds tightly to eIF4E and sequesters it, thus preventing its association with eIF4G. The HAV IRES was also inhibited by addition of m(7)GpppG cap analogue, irrespective of whether the RNA tested was capped or not. Thus, initiation on the HAV IRES requires that eIF4E be associated with eIF4G and that the cap-binding pocket of eIF4E be empty and unoccupied. This suggests two alternative models: (i) initiation requires a direct interaction between an internal site in the IRES and eIF4E/4G, an interaction which involves the cap-binding pocket of eIF4E in addition to any direct eIF4G-RNA interactions; or (ii) it requires eIF4G in a particular conformation which can be attained only if eIF4E is bound to it, with the cap binding pocket of the eIF4E unoccupied. PMID- 11483730 TI - Detailed analysis of the requirements of hepatitis A virus internal ribosome entry segment for the eukaryotic initiation factor complex eIF4F. AB - The hepatitis A virus (HAV) internal ribosome entry segment (IRES) is unique among the picornavirus IRESs in that it is inactive in the presence of either the entero- and rhinovirus 2A or aphthovirus Lb proteinases. Since these proteinases both cleave eukaryotic initiation factor 4G (eIF4G) and HAV IRES activity could be rescued in vitro by addition of eIF4F to proteinase-treated extracts, it was concluded that the HAV IRES requires eIF4F containing intact eIF4G. Here, we show that the inability of the HAV IRES to function with cleaved eIF4G cannot be attributed to inefficient binding of the cleaved form of eIF4G by the HAV IRES. Indeed, the binding of both intact eIF4F and the C-terminal cleavage product of eIF4G to the HAV IRES was virtually indistinguishable from their binding to the encephalomyocarditis virus IRES, as assessed by UV cross-linking and filter retention assays. Rather, we show that HAV IRES activity requires, either directly or indirectly, components of the eIF4F complex which interact with the N terminal fragment of eIF4G. Effectively, HAV IRES activity, but not that of the human rhinovirus IRES, was sensitive to the rotavirus nonstructural protein NSP3 [which displaces poly(A)-binding protein from the eIF4F complex], to recombinant eIF4E-binding protein (which prevents the association of the cap binding protein eIF4E with eIF4G), and to cap analogue. PMID- 11483731 TI - Copper binding to the PrP isoforms: a putative marker of their conformation and function. AB - We show here that PrP(C), the normal isoform of the prion protein (PrP(Sc)), could be retained by a Cu(2+)-loaded resin through two different binding sites. Contrarily, PrP(Sc) was not retained at all by such resin. This constitutes a new prion-specific property of PrP(Sc), which in addition to protease resistance and beta-sheet content, may result from its aberrant conformation. PMID- 11483732 TI - Influenza virus ns1 protein induces apoptosis in cultured cells. AB - The importance of influenza viruses as worldwide pathogens in humans, domestic animals, and poultry is well recognized. Discerning how influenza viruses interact with the host at a cellular level is crucial for a better understanding of viral pathogenesis. Influenza viruses induce apoptosis through mechanisms involving the interplay of cellular and viral factors that may depend on the cell type. However, it is unclear which viral genes induce apoptosis. In these studies, we show that the expression of the nonstructural (NS) gene of influenza A virus is sufficient to induce apoptosis in MDCK and HeLa cells. Further studies showed that the multimerization domain of the NS1 protein but not the effector domain is required for apoptosis. However, this mutation is not sufficient to inhibit apoptosis using whole virus. PMID- 11483733 TI - DNA binding and modulation of gene expression by the latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. The latency-associated nuclear antigen (LANA) is highly expressed in these malignancies and has been shown to play an important role in episomal maintenance, presumably by binding to a putative oriP. In addition, LANA modulates cellular and viral gene expression and interacts with the cellular tumor suppressors p53 and retinoblastoma suppressor protein. Many of these features are reminiscent of Epstein-Barr virus nuclear antigens (EBNAs), a family of six proteins expressed during latency. EBNA-1 is required for episome maintenance, binds to oriP, and strongly activates transcription from two promoters, including its own. We have previously shown that LANA can transactivate its own promoter and therefore asked whether LANA, like EBNA-1, activates transcription by direct binding to DNA. By using recombinant LANA expressed from vaccinia virus vectors for electrophoretic mobility shift assays, we found that LANA does not bind to its own promoter. In contrast, LANA binds specifically to sequences containing an imperfect 20-bp palindrome in the terminal repeat (TR) of KSHV. We further show that the C-terminal domain of LANA is sufficient for site-specific DNA binding. Unlike EBNA-1, which activates transcription through binding of oriP, we found that LANA inhibits transcription from a single TR binding site. A multimerized TR as found in the viral genome results in strong transcriptional suppression when linked to a heterologous promoter. These data suggest that LANA, although fulfilling functions similar to those of EBNA-1, does so by very different mechanisms. PMID- 11483734 TI - The envelope glycoprotein of friend spleen focus-forming virus covalently interacts with and constitutively activates a truncated form of the receptor tyrosine kinase Stk. AB - The Friend spleen focus-forming virus (SFFV) encodes a unique envelope glycoprotein, gp55, which allows erythroid cells to proliferate and differentiate in the absence of erythropoietin (Epo). SFFV gp55 has been shown to interact with the Epo receptor complex, causing constitutive activation of various signal transducing molecules. When injected into adult mice, SFFV induces a rapid erythroleukemia, with susceptibility being determined by the host gene Fv-2, which was recently shown to be identical to the gene encoding the receptor tyrosine kinase Stk/Ron. Susceptible, but not resistant, mice encode not only full-length Stk but also a truncated form of the kinase, sf-Stk, which may mediate the biological effects of SFFV infection. To determine whether expression of SFFV gp55 leads to the activation of sf-Stk, we expressed sf-Stk, with or without SFFV gp55, in hematopoietic cells expressing the Epo receptor. Our data indicate that sf-Stk interacts with SFFV gp55 as well as gp55(P), the biologically active form of the viral glycoprotein, forming disulfide-linked complexes. This covalent interaction, as well as noncovalent interactions with SFFV gp55, results in constitutive tyrosine phosphorylation of sf-Stk and its association with multiple tyrosine-phosphorylated signal-transducing molecules. In contrast, neither Epo stimulation in the absence of SFFV gp55 expression nor expression of a mutant of SFFV that cannot interact with sf-Stk was able to induce tyrosine phosphorylation of sf-Stk or its association with any signal transducing molecules. Covalent interaction of sf-Stk with SFFV gp55 and constitutive tyrosine phosphorylation of sf-Stk can also be detected in an erythroleukemia cell line derived from an SFFV-infected mouse. Our results suggest that SFFV gp55 may mediate its biological effects in vivo by interacting with and activating a truncated form of the receptor tyrosine kinase Stk. PMID- 11483735 TI - Posttranslational processing of infected cell proteins 0 and 4 of herpes simplex virus 1 is sequential and reflects the subcellular compartment in which the proteins localize. AB - The herpes simplex virus 1 (HSV-1) infected cell proteins 0 and 4 (ICP0 and ICP4) are multifunctional proteins extensively posttranscriptionally processed by both cellular and viral enzymes. We examined by two-dimensional separations the posttranslational forms of ICP0 and ICP4 in HEp-2 cells and in human embryonic lung (HEL) fibroblasts infected with wild-type virus, mutant R325, lacking the sequences encoding the U(S)1.5 protein and the overlapping carboxyl-terminal domain of ICP22, or R7914, in which the aspartic acid 199 of ICP0 was replaced by alanine. We report the following (i) Both ICP0 and ICP4 were sequentially posttranslationally modified at least until 12 h after infection. In HEL fibroblasts, the processing of ICP0 shifted from A+B forms at 4 h to D+G forms at 8 h and finally to G, E, and F forms at 12 h. The ICP4 progression was from the A' form noted at 2 h to B' and C' forms noted at 4 h to the additional D' and E' forms noted at 12 h. The progression tended to be toward more highly charged forms of the proteins. (ii) Although the overall patterns were similar, the mobility of proteins made in HEp-2 cells differed from those made in HEL fibroblasts. (iii) The processing of ICP0 forms E and F was blocked in HEL fibroblasts infected with R325 or with wild-type virus and treated with roscovitine, a specific inhibitor of cell cycle-dependent kinases cdc2, cdk2, and cdk5. R325-infected HEp-2 cells lacked the D' form of ICP4, and roscovitine blocked the appearance of the most highly charged E' form of ICP4. (iv) A characteristic of ICP0 is that it is translocated into the cytoplasm of HEL fibroblasts between 5 and 9 h after infection. Addition of MG132 to the cultures late in infection resulted in rapid relocation of cytoplasmic ICP0 back into the nucleus. Exposure of HEL fibroblasts to MG132 late in infection resulted in the disappearance of the highly charged ICP0 G isoform. The G form of ICP0 was also absent in cells infected with R7914 mutant. In cells infected with this mutant, ICP0 is not translocated to the cytoplasm. (v) Last, cdc2 was active in infected cells, and this activity was inhibited by roscovitine. In contrast, the activity of cdk2 exhibited by immunoprecipitated protein was reduced and resistant to roscovitine and may represent a contaminating kinase activity. We conclude from these results that the ICP0 G isoform is the cytoplasmic form, that it may be phosphorylated by cdc2, consistent with evidence published earlier (S. J., Advani, R. R. Weichselbaum, and B. Roizman, Proc. Natl. Acad. Sci. USA 96:10996 11001, 2000), and that the processing is reversed upon relocation of the G isoform from the cytoplasm into the nucleus. The processing of ICP4 is also affected by R325 and roscovitine. The latter result suggests that ICP4 may also be a substrate of cdc2 late in infection. Last, additional modifications are superimposed by cell-type-specific enzymes. PMID- 11483736 TI - Multimerization of human immunodeficiency virus type 1 Gag promotes its localization to barges, raft-like membrane microdomains. AB - The Gag polyprotein of human immunodeficiency virus type 1 (HIV-1) organizes the assembly of nascent virions at the plasma membrane of infected cells. Here we demonstrate that a population of Gag is present in distinct raft-like membrane microdomains that we have termed "barges." Barges have a higher density than standard rafts, most likely due to the presence of oligomeric Gag-Gag assembly complexes. The regions of the Gag protein responsible for barge targeting were mapped by examining the flotation behavior of wild-type and mutant proteins on Optiprep density gradients. N-myristoylation of Gag was necessary for association with barges. Removal of the NC and p6 domains shifted much of the Gag from barges into typical raft fractions. These data are consistent with a model in which multimerization of myristoylated Gag proteins drives association of Gag oligomers into raft-like barges. The functional significance of barge association was revealed by several lines of evidence. First, Gag isolated from virus-like particles was almost entirely localized in barges. Moreover, a comparison of wild type Gag with Fyn(10)Gag, a chimeric protein containing the N-terminal sequence of Fyn, revealed that Fyn(10)Gag exhibited increased affinity for barges and a two- to fourfold increase in particle production. These results imply that association of Gag with raft-like barge membrane microdomains plays an important role in the HIV-1 assembly process. PMID- 11483737 TI - Highly productive infection with pseudotyped human immunodeficiency virus type 1 (HIV-1) indicates no intracellular restrictions to HIV-1 replication in primary human astrocytes. AB - Human astrocytes can be infected with human immunodeficiency virus type 1 (HIV-1) in vitro and in vivo, but, in contrast to T lymphocytes and macrophages, virus expression is inefficient. To investigate the HIV-1 life cycle in human fetal astrocytes, we infected cells with HIV-1 pseudotyped with envelope glycoproteins of either amphotropic murine leukemia virus or vesicular stomatitis virus. Infection by both pseudotypes was productive and long lasting and reached a peak of 68% infected cells and 1.7 microg of viral p24 per ml of culture supernatant 7 days after virus inoculation and then continued with gradually declining levels of virus expression through 7 weeks of follow-up. This contrasted with less than 0.1% HIV-1 antigen-positive cells and 400 pg of extracellular p24 per ml at the peak of astrocyte infection with native HIV-1. Cell viability and growth kinetics were similar in infected and control cells. Northern blot analysis revealed the presence of major HIV-1 RNA species of 9, 4, and 2 kb in astrocytes exposed to pseudotyped (but not wild-type) HIV-1 at 2, 14, and 28 days after infection. Consistent with productive infection, the 9- and 4-kb viral transcripts in astrocytes infected by pseudotyped HIV-1 were as abundant as the 2-kb mRNA during 4 weeks of follow-up, and both structural and regulatory viral proteins were detected in infected cells by immunoblotting or cell staining. The progeny virus released by these cells was infectious. These results indicate that the major barrier to HIV-1 infection of primary astrocytes is at virus entry and that astrocytes have no intrinsic intracellular restriction to efficient HIV-1 replication. PMID- 11483738 TI - Mutations in the fusion peptide and adjacent heptad repeat inhibit folding or activity of the Newcastle disease virus fusion protein. AB - Paramyxovirus fusion proteins have two heptad repeat domains, HR1 and HR2, which have been implicated in the fusion activity of the protein. Peptides with sequences from these two domains form a six-stranded coiled coil, with the HR1 sequences forming a central trimer (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999; X. Zhao, M. Singh, V. N. Malashkevich, and P. S. Kim, Proc. Natl. Acad. Sci. USA 97:14172-14177, 2000). We have extended our previous mutational analysis of the HR1 domain of the Newcastle disease virus fusion protein, focusing on the role of the amino acids forming the hydrophobic core of the trimer, amino acids in the "a" and "d" positions of the helix from amino acids 123 to 182. Both conservative and nonconservative point mutations were characterized for their effects on synthesis, stability, proteolytic cleavage, and surface expression. Mutant proteins expressed on the cell surface were characterized for fusion activity by measuring syncytium formation, content mixing, and lipid mixing. We found that all mutations in the "a" position interfered with proteolytic cleavage and surface expression of the protein, implicating the HR1 domain in the folding of the F protein. However, mutation of five of seven "d" position residues had little or no effect on surface expression but, with one exception at residue 175, did interfere to various extents with the fusion activity of the protein. One of these "d" mutations, at position 154, interfered with proteolytic cleavage, while the rest of the mutants were cleaved normally. That most "d" position residues do affect fusion activity argues that a stable HR1 trimer is required for formation of the six-stranded coiled coil and, therefore, optimal fusion activity. That most of the "d" position mutations do not block folding suggests that formation of the core trimer may not be required for folding of the prefusion form of the protein. We also found that mutations within the fusion peptide, at residue 128, can interfere with folding of the protein, implicating this region in folding of the molecule. No characterized mutation enhanced fusion. PMID- 11483739 TI - Human immunodeficiency virus type 1 replication in the absence of integrase mediated dna recombination: definition of permissive and nonpermissive T-cell lines. AB - Functional retroviral integrase protein is thought to be essential for productive viral replication. Yet, previous studies differed on the extent to which integrase mutant viruses expressed human immunodeficiency virus type 1 (HIV-1) genes from unintegrated DNA. Although one reason for this difference was that class II integrase mutations pleiotropically affected the viral life cycle, another reason apparently depended on the identity of the infected cell. Here, we analyzed integrase mutant viral infectivities in a variety of cell types. Single round infectivity of class I integration-specific mutant HIV-1 ranged from <0.03 to 0.3% of that of the wild type (WT) across four different T-cell lines. Based on this approximately 10-fold influence of cell type on mutant gene expression, we examined class I and class II mutant replication kinetics in seven different cell lines and two primary cell types. Unexpectedly, some cell lines supported productive class I mutant viral replication under conditions that restricted class II mutant growth. Cells were defined as permissive, semipermissive, or nonpermissive based on their ability to support the continual passage of class I integration-defective HIV-1. Mutant infectivity in semipermissive and permissive cells as quantified by 50% tissue culture infectious doses, however, was only 0.0006 to 0.005% of that of WT. Since the frequencies of mutant DNA recombination in these lines ranged from 0.023 to <0.093% of the WT, we conclude that productive replication in the absence of integrase function most likely required the illegitimate integration of HIV-1 into host chromosomes by cellular DNA recombination enzymes. PMID- 11483740 TI - Serum and mucosal immune responses to an inactivated influenza virus vaccine induced by epidermal powder immunization. AB - Both circulating and mucosal antibodies are considered important for protection against infection by influenza virus in humans and animals. However, current inactivated vaccines administered by intramuscular injection using a syringe and needle elicit primarily circulating antibodies. In this study, we report that epidermal powder immunization (EPI) via a unique powder delivery system elicits both serum and mucosal antibodies to an inactivated influenza virus vaccine. Serum antibody responses to influenza vaccine following EPI were enhanced by codelivery of cholera toxin (CT), a synthetic oligodeoxynucleotide containing immunostimulatory CpG motifs (CpG DNA), or the combination of these two adjuvants. In addition, secretory immunoglobulin A (sIgA) antibodies were detected in the saliva and mucosal lavages of the small intestine, trachea, and vaginal tract, although the titers were much lower than the IgG titers. The local origin of the sIgA antibodies was further shown by measuring antibodies released from cultured tracheal and small intestinal fragments and by detecting antigen specific IgA-secreting cells in the lamina propria using ELISPOT assays. EPI with a single dose of influenza vaccine containing CT or CT and CpG DNA conferred complete protection against lethal challenges with an influenza virus isolated 30 years ago, whereas a prime and boost immunizations were required for protection in the absence of an adjuvant. The ability to elicit augmented circulating antibody and mucosal antibody responses makes EPI a promising alternative to needle injection for administering vaccines against influenza and other diseases. PMID- 11483741 TI - Simultaneous positive and purifying selection on overlapping reading frames of the tat and vpr genes of simian immunodeficiency virus. AB - Tat-specific cytotoxic T cells have previously been shown to exert positive Darwinian selection favoring amino acid replacements of an epitope of simian immunodeficiency virus (SIV). The region of the tat gene encoding this epitope falls within a region of overlap between the tat and vpr reading frames, and nonsynonymous nucleotide substitutions in the tat reading frame were found to occur disproportionately in such a way as to cause synonymous changes in the vpr reading frame. Comparison of published complete SIV genomes showed Tat to be the least conserved at the amino acid level of nine proteins encoded by the virus, while Vpr was one of the most conserved. Numerous parallel amino acid changes occurred within the Tat epitope independently in different monkeys, and purifying selection on the vpr reading frame, by limiting acceptable nonsynonymous substitutions in the tat reading frame, evidently has enhanced the probability of parallel evolution. PMID- 11483742 TI - Human immunodeficiency virus type 1 DNA sequences genetically damaged by hypermutation are often abundant in patient peripheral blood mononuclear cells and may be generated during near-simultaneous infection and activation of CD4(+) T cells. AB - G-to-A hypermutation has been sporadically observed in human immunodeficiency virus type 1 (HIV-1) proviral sequences from patient peripheral blood mononuclear cells (PBMC) and virus cultures but has not been systematically evaluated. PCR primers matched to normal and hypermutated sequences were used in conjunction with an agarose gel electrophoresis system incorporating an AT-binding dye to visualize, separate, clone, and sequence hypermutated and normal sequences in the 297-bp HIV-1 protease gene amplified from patient PBMC. Among 53 patients, including individuals infected with subtypes A through D and at different clinical stages, at least 43% of patients harbored abundant hypermutated, along with normal, protease genes. In 70 hypermutated sequences, saturation of G residues in the GA or GG dinucleotide context ranged from 20 to 94%. Levels of other mutants were not elevated, and G-to-A replacement was entirely restricted to GA or GG, and not GC or GT, dinucleotides. Sixty-nine of 70 hypermutated and 3 of 149 normal sequences had in-frame stop codons. To investigate the conditions under which hypermutation occurs in cell cultures, purified CD4(+) T cells from normal donors were infected with cloned NL4-3 virus stocks at various times before and after phytohemagglutinin (PHA) activation. Hypermutation was pronounced when HIV-1 infection occurred simultaneously with, or a few hours after, PHA activation, but after 12 h or more after PHA activation, most HIV-1 sequences were normal. Hypermutated sequences generated in culture corresponded exactly in all parameters to those obtained from patient PBMC. Near-simultaneous activation and infection of CD4(+) T cells may represent a window of susceptibility where the informational content of HIV-1 sequences is lost due to hypermutation. PMID- 11483743 TI - Chimeric nectin1-poliovirus receptor molecules identify a nectin1 region functional in herpes simplex virus entry. AB - Human nectin1 (hNectin1), an adhesion molecule belonging to the nectin family of the immunoglobulin superfamily, mediates entry of herpes simplex virus (HSV) into cells. The hNectin1 domain that mediates virus entry into cells and also binds glycoprotein D (gD) has been localized to the first N-terminal V-type domain. The poliovirus receptor (PVR) is a structural homolog to nectins, but it cannot function as an HSV entry receptor. hNectin1-PVR chimeras were constructed to functionally locate the site on hNectin1 involved in HSV entry (HSV entry site). The epitope recognized by monoclonal antibody (MAb) R1.302, which is able to block HSV entry, was also located. The chimeric receptors were designed to preserve the overall structure of the V domain. The HSV entry activity mapped entirely to the hNectin1 portion located between residues 64 and 94 (64-94), likely to encode the C, C', and C" beta-strands and intervening loops. In turn, this site consisted of two portions: one with low-level basal activity for HSV entry (77-94), and one immediately upstream (residues 64 to 76) which greatly enhanced the HSV entry activity of the downstream region. The gD-binding site mapped substantially to the same site, whereas the MAb R1.302 epitope also required a further downstream portion (95-102). The involvement of the 64-76 portion is at difference with previous indirect mapping results that were based on competitive binding studies (C. Krummenacher et al., J. Virol. 74:10863-10872, 2000). The A, A', B, D, E, F, and G beta-strands and intervening loops did not appear to play any role in HSV entry. According to the predicted three dimensional structure of PVR, the C C' C" site is located peripherally in the V domain and very likely represents an accessible portion at the cell surface. PMID- 11483744 TI - Specific interaction of a novel foamy virus Env leader protein with the N terminal Gag domain. AB - Cryoelectron micrographs of purified human foamy virus (HFV) and feline foamy virus (FFV) particles revealed distinct radial arrangements of Gag proteins. The capsids were surrounded by an internal Gag layer that in turn was surrounded by, and separated from, the viral membrane. The width of this layer was about 8 nm for HFV and 3.8 nm for FFV. This difference in width is assumed to reflect the different sizes of the HFV and FFV MA domains: the HFV MA domain is about 130 residues longer than that of FFV. The distances between the MA layer and the edge of the capsid were identical in different particle classes. In contrast, only particles with a distended envelope displayed an invariant, close spacing between the MA layer and the Env membrane which was absent in the majority of particles. This indicates a specific interaction between MA and Env at an unknown step of morphogenesis. This observation was supported by surface plasmon resonance studies. The purified N-terminal domain of FFV Gag specifically interacted with synthetic peptides and a defined protein domain derived from the N-terminal Env leader protein. The specificity of this interaction was demonstrated by using peptides varying in the conserved Trp residues that are known to be required for HFV budding. The interaction with Gag required residues within the novel virion associated FFV Env leader protein of about 16.5 kDa. PMID- 11483745 TI - Requirements for the induction of interleukin-6 by herpes simplex virus-infected leukocytes. AB - Cytokines play important roles in the clearance of herpes simplex virus (HSV) infections and in virus-induced immunopathology. One cytokine known to contribute to resistance against HSV is interleukin-6 (IL-6). Here we have investigated virus-cell interactions responsible for IL-6 induction by HSV in leukocytes. Both HSV type 1 and type 2 are potent inducers of IL-6, and this phenomenon is augmented in the presence of gamma interferon. The ability to induce IL-6 is dependent on de novo protein synthesis and is sensitive to UV irradiation of the virus. Virus mutants lacking the virion-transactivating protein VP16 or any of the immediate-early proteins ICP0, ICP4, or ICP27 displayed unaltered capacities to induce IL-6. However, wild-type virus was unable to induce IL-6 in a macrophage cell line overexpressing a mutant of double-stranded RNA-activated protein kinase (PKR). This suggests a role for PKR in HSV-induced IL-6 expression. HSV infection led to enhanced binding to the kappaB, CRE, and AP-1 sites of the IL-6 promoter, and inhibitors against NF-kappaB and the p38 kinase strongly reduced accumulation of IL-6 mRNA in infected cells. Moreover, macrophage cell lines expressing dominant negative mutants of IkappaBalpha and p38 responded to HSV-1 infection with reduced IL-6 expression compared to the control-vector-transfected cell line. The results show that induction of IL-6 by HSV in leukocytes is dependent on PKR and cellular signaling through NF-kappaB and a p38-dependent pathway. PMID- 11483746 TI - Antibody-directed targeting of retroviral vectors via cell surface antigens. AB - Targeted stable transduction of specific cells is a highly desirable goal for gene therapy applications. We report an efficient and broadly applicable approach for targeting retroviral vectors to specific cells. We find that the envelope of the alphavirus Sindbis virus can pseudotype human immunodeficiency virus type 1- and murine leukemia virus-based retroviral vectors. When modified to contain the Fc-binding domain of protein A, this envelope gives a significant enhancement in specificity in combination with antibodies specific for HLA and CD4 relative to that without antibody. Unlike previous targeting strategies for retroviral transduction, the virus titers are relatively high and stable and can be further increased by ultracentrifugation. This study provides proof of principle for a targeting strategy that would be generally useful for many gene therapy applications. PMID- 11483747 TI - Construction of an infectious cDNA clone of Aichi virus (a new member of the family Picornaviridae) and mutational analysis of a stem-loop structure at the 5' end of the genome. AB - Aichi virus is the type species of a new genus, Kobuvirus, of the family Picornaviridae. In this study, we constructed a full-length cDNA clone of Aichi virus whose in vitro transcripts were infectious to Vero cells. During construction of the infectious cDNA clone, a novel sequence of 32 nucleotides was identified at the 5' end of the genome. Computer-assisted prediction of the secondary structure of the 5' end of the genome, including the novel sequence, suggested the formation of a stable stem-loop structure consisting of 42 nucleotides. The function of this stem-loop in virus replication was investigated using various site-directed mutants derived from the infectious cDNA clone. Our data indicated that correct folding of the stem-loop at the 5' end of the positive strand, but not at the 3' end of the negative strand, is critical for viral RNA replication. The primary sequence in the lower part of the stem was also suggested to be crucial for RNA replication. In contrast, nucleotide changes in the loop segment did not so severely reduce the efficiency of virus replication. A double mutant, in which both nucleotide stretches of the middle part of the stem were replaced by their complementary nucleotides, had efficient RNA replication and translation abilities but was unable to produce viruses. These results indicate that the stem-loop at the 5' end of the Aichi virus genome is an element involved in both viral RNA replication and production of infectious virus particles. PMID- 11483748 TI - The arginine-1493 residue in QRRGRTGR1493G motif IV of the hepatitis C virus NS3 helicase domain is essential for NS3 protein methylation by the protein arginine methyltransferase 1. AB - The NS3 protein of hepatitis C virus (HCV) contains protease and RNA helicase activities, both of which are likely to be essential for HCV propagation. An arginine residue present in the arginine-glycine (RG)-rich region of many RNA binding proteins is posttranslationally methylated by protein arginine methyltransferases (PRMTs). Amino acid sequence analysis revealed that the NS3 protein contains seven RG motifs, including two potential RG motifs in the 1486 QRRGRTGRG-1494 motif IV of the RNA helicase domain, in which arginines are potentially methylated by PRMTs. Indeed, we found that the full-length NS3 protein is arginine methylated in vivo. The full-length NS3 protein and the NS3 RNA helicase domain were methylated by a crude human cell extract. The purified PRMT1 methylated the full-length NS3 and the RNA helicase domain, but not the NS3 protease domain. The NS3 helicase bound specifically and comigrated with PRMT1 in vitro. Mutational analyses indicate that the Arg(1493) in the QRR(1488)GRTGR(1493)G region of the NS3 RNA helicase is essential for NS3 protein methylation and that Arg(1488) is likely methylated. NS3 protein methylation by the PRMT1 was decreased in the presence of homoribopolymers, suggesting that the arginine-rich motif IV is involved in RNA binding. The results suggest that an arginine residue(s) in QRXGRXGR motif IV conserved in the virus-encoded RNA helicases can be posttranslationally methylated by the PRMT1. PMID- 11483749 TI - Conversion in the requirement of coat protein in cell-to-cell movement mediated by the cucumber mosaic virus movement protein. AB - Plant viruses have movement protein (MP) gene(s) essential for cell-to-cell movement in hosts. Cucumber mosaic virus (CMV) requires its own coat protein (CP) in addition to the MP for intercellular movement. Our present results using variants of both CMV and a chimeric Brome mosaic virus with the CMV MP gene revealed that CMV MP truncated in its C-terminal 33 amino acids has the ability to mediate viral movement independently of CP. Coexpression of the intact and truncated CMV MPs extremely reduced movement of the chimeric viruses, suggesting that these heterogeneous CMV MPs function antagonistically. Sequential deletion analyses of the CMV MP revealed that the dispensability of CP occurred when the C terminal deletion ranged between 31 and 36 amino acids and that shorter deletion impaired the ability of the MP to promote viral movement. This is the first report that a region of MP determines the requirement of CP in cell-to-cell movement of a plant virus. PMID- 11483750 TI - Genetic variation of Citrus tristeza virus isolates from California and Spain: evidence for mixed infections and recombination. AB - We examined the population structure and genetic variation of four genomic regions within and between 30 Citrus tristeza virus (CTV) isolates from Spain and California. Our analyses showed that most isolates contained a population of sequence variants, with one being predominant. Four isolates showed two major sequence variants in some genomic regions. The two major variants of three of these isolates showed very low nucleotide identity to each other but were very similar to those of other isolates, suggesting the possibility of mixed infections with two divergent isolates. Incongruencies of phylogenetic relationships in the different genomic regions and statistical analyses suggested that the genomes of some CTV sequence variants originated by recombination events between diverged sequence variants. No correlation was observed between geographic origin and nucleotide distance, and thus from a genetic view, the Spanish and Californian isolates analyzed here could be considered members of the same population. PMID- 11483751 TI - Susceptibility of rat-derived cells to replication by human immunodeficiency virus type 1. AB - Progress in developing a small animal model of human immunodeficiency virus type 1 (HIV-1) disease would greatly facilitate studies of transmission, pathogenesis, host immune responses, and antiviral strategies. In this study, we have explored the potential of rats as a susceptible host. In a single replication cycle, rat cell lines Rat2 and Nb2 produced infectious virus at levels 10- to 60-fold lower than those produced by human cells. Rat-derived cells supported substantial levels of early HIV-1 gene expression, which was further enhanced by overexpression of human cyclin T1. Rat cells displayed quantitative, qualitative, and cell-type-specific limitations in the late phase of the HIV-1 replication cycle including relative expression levels of HIV-1 Gag proteins, intracellular Gag processing, and viral egress. Nb2 cells were rendered permissive to HIV-1 R5 viruses by coexpression of human CD4 and CCR5, indicating that the major restriction on HIV-1 replication was at the level of cellular entry. We also found that primary rat lymphocytes, macrophages, and microglia expressed considerable levels of early HIV-1 gene products following infection with pseudotyped HIV-1. Importantly, primary rat macrophages and microglia, but not lymphocytes, also expressed substantial levels of HIV-1 p24 CA and produced infectious virions. Collectively, these results identify the rat as a promising candidate for a transgenic small animal model of HIV-1 infection and highlight pertinent cell-type-specific restrictions that are features of this species. PMID- 11483752 TI - Ribavirin induces error-prone replication of GB virus B in primary tamarin hepatocytes. AB - GB virus B (GBV-B) is the closest relative of hepatitis C virus (HCV) and is an attractive surrogate model for HCV antiviral studies. GBV-B induces an acute, resolving hepatitis in tamarins. Utilizing primary cultures of tamarin hepatocytes, we have previously developed a tissue culture system that exhibits high levels of GBV-B replication. In this report, we have extended the utility of this system for testing antiviral compounds. Treatment with human interferon provided only a marginal antiviral effect, while poly(I-C) yielded >3 and 4 log units of reduction of cell-associated and secreted viral RNA, respectively. Interestingly, treatment of GBV-B-infected hepatocytes with ribavirin resulted in an approximately 4-log decrease in viral RNA levels. Guanosine blocked the antiviral effect of ribavirin, suggesting that inhibition of IMP dehydrogenase (IMPDH) and reduction of intracellular GTP levels were essential for the antiviral effect. However, mycophenolic acid, another IMPDH inhibitor, had no antiviral effect. Virions harvested from ribavirin-treated cultures exhibited a dramatically reduced specific infectivity. These data suggest that incorporation of ribavirin triphosphate induces error-prone replication with concomitant reduction in infectivity and that reduction of GTP pools may be required for incorporation of ribavirin triphosphate. In contrast to the in vitro studies, no significant reduction in viremia was observed in vivo following treatment of tamarins with ribavirin during acute infection with GBV-B. These findings are consistent with the observation that ribavirin monotherapy for HCV infection decreases liver disease without a significant reduction in viremia. Our data suggest that nucleoside analogues that induce error-prone replication could be an attractive approach for the treatment of HCV infection if administered at sufficient levels to result in efficient incorporation by the viral polymerase. PMID- 11483753 TI - Role of the proline-rich motif of bovine leukemia virus transmembrane protein gp30 in viral load and pathogenicity in sheep. AB - The cytoplasmic tail of bovine leukemia virus (BLV) transmembrane protein gp30 has multiple amino acid motifs that mimic those present in signaling proteins associated with B-cell and T-cell receptors. The proline-rich motif of gp30, PX(2)PX(4-5)P, is analogous to the recognition site of Src homology 3 (SH3) domains of signaling molecules. Using site-directed mutagenesis of an infectious molecular clone of BLV, point mutations were introduced which changed three of the prolines of the motif to alanines. The influence of these mutations on the pathogenicity of BLV was studied in sheep which received either (i) plasmid DNA with provirus containing proline-to-alanine mutations (pppBLV), (ii) plasmid DNA with wild-type provirus (wtBLV), or (iii) transfection reagent alone. Although all of the BLV-injected animals seroconverted at approximately the same time, viral loads at later time points were high in five of five of the wtBLV group and two of five of the pppBLV group but low in three of five of the pppBLV group, as determined by semiquantitative PCR. Viral expression was lower in the pppBLV transfected sheep, as measured by p24 antigen enzyme-linked immunosorbent assay in cultured cells, and serologic titers were lower. Thirty-one months after transfection, four of four wtBLV-transfected sheep had died of leukemia and lymphoma, and all five of the pppBLV-transfected sheep were clinically healthy and had normal peripheral blood lymphocyte counts. These data indicate that the proline-rich motif of gp30 is not required for viral infectivity but is important for high viral load in vivo, suggesting that SH3-mediated gp30 interactions are critical for viral pathogenesis following infection. Absence of interactions with the proline-rich motif may prevent or delay tumorigenesis in sheep. PMID- 11483754 TI - In vitro activation of feline immunodeficiency virus in ramified microglial cells from asymptomatically infected cats. AB - Intravenous infection of cats with feline immunodeficiency virus was used as a model system to study activation of virus replication in brain-resident microglial cells in vitro. Virus release by ramified microglial cells isolated from subclinically infected animals was detectable in cell-free tissue culture supernatant only by reverse transcription and nested PCR of gag-specific RNA sequences and not by virion-associated reverse transcriptase activity. In contrast, cocultivation of in vivo-infected microglial cells with mitogen activated peripheral blood mononuclear cells (PBMC) regularly allows detection of high virus yields in cell-free tissue culture fluid. Besides uptake and multiplication of microglia-derived virus in PBMC, release of virus from microglia is stimulated by cell contact with PBMC. The data suggest that T lymphocytes patrolling the central nervous system could reactivate the semilatent state of lentiviruses in microglial cells in the course of clinically silent central nervous system infection. PMID- 11483755 TI - Host switching in Lyssavirus history from the Chiroptera to the Carnivora orders. AB - Lyssaviruses are unsegmented RNA viruses causing rabies. Their vectors belong to the Carnivora and Chiroptera orders. We studied 36 carnivoran and 17 chiropteran lyssaviruses representing the main genotypes and variants. We compared their genes encoding the surface glycoprotein, which is responsible for receptor recognition and membrane fusion. The glycoprotein is the main protecting antigen and bears virulence determinants. Point mutation is the main force in lyssavirus evolution, as Sawyer's test and phylogenetic analysis showed no evidence of recombination. Tests of neutrality indicated a neutral model of evolution, also supported by globally high ratios of synonymous substitutions (d(S)) to nonsynonymous substitutions (d(N)) (>7). Relative-rate tests suggested similar rates of evolution for all lyssavirus lineages. Therefore, the absence of recombination and similar evolutionary rates make phylogeny-based conclusions reliable. Phylogenetic reconstruction strongly supported the hypothesis that host switching occurred in the history of lyssaviruses. Indeed, lyssaviruses evolved in chiropters long before the emergence of carnivoran rabies, very likely following spillovers from bats. Using dated isolates, the average rate of evolution was estimated to be roughly 4.3 x 10(-4) d(S)/site/year. Consequently, the emergence of carnivoran rabies from chiropteran lyssaviruses was determined to have occurred 888 to 1,459 years ago. Glycoprotein segments accumulating more d(N) than d(S) were distinctly detected in carnivoran and chiropteran lyssaviruses. They may have contributed to the adaptation of the virus to the two distinct mammal orders. In carnivoran lyssaviruses they overlapped the main antigenic sites, II and III, whereas in chiropteran lyssaviruses they were located in regions of unknown functions. PMID- 11483756 TI - RNAs extracted from herpes simplex virus 1 virions: apparent selectivity of viral but not cellular RNAs packaged in virions. AB - Following the lead of recent studies on the presence of RNA in virions of human cytomegalovirus, we investigated the presence and identity of RNAs from purified virions of herpes simple virus 1. To facilitate these studies, we designed primers for all known open reading frames (ORFs) and also constructed cDNA arrays containing probes designed to detect all known transcripts. In the first series of experiments, labeled DNA made by reverse transcription of poly(A)(+) RNA extracted from infected HEp-2 or rabbit skin cells hybridized to all but two of the probes in the cDNA array. A similar analysis of the RNA extracted from purified extracellular virions derived from infected HEp-2 cells hybridized to probes representing 24 of the ORFs. In the second series of analyses, we reverse transcribed and amplified by PCR RNAs from purified intracellular or extracellular virions derived from infected HEp-2 or Vero cell lines. The positive RNAs were retested by PCR with and without prior reverse transcription to ensure that the samples tested were free of contaminating DNA. The results were as follows. (i) Only a fraction of viral ORF transcripts were represented in virion RNA, and only nine RNAs (U(L)10, U(L)34/U(L)35, U(L)36, U(L)42, U(L)48, U(L)51, U(S)1/U(S)1.5, U(S)8.5, and U(S)10/U(S)11) were positive in all RT PCR assays. Of these, seven were positive by hybridization to cDNA arrays. (ii) RNA extracted from cells infected with a mutant virus lacking the U(S)8 to U(S)12 genes yielded results similar to those described above, indicating that U(S)11, a known RNA binding protein, does not play a role in packaging RNA in virions. (iii) Cellular RNAs detected in virions were representative of the abundant cellular RNAs. Last, RNA extracted from virions was translated in vitro and the translation products were reacted with antibody to alphaTIF (VIP16). The immune precipitate contained a labeled protein with the apparent molecular weight of alphaTIF, indicating that at least one mRNA packaged in virions was intact and capable of being translated. The basis for the apparent selectivity in the packaging of the viral RNAs packaged in virions is unknown. PMID- 11483757 TI - Use of whole genome sequence data to infer baculovirus phylogeny. AB - Several phylogenetic methods based on whole genome sequence data were evaluated using data from nine complete baculovirus genomes. The utility of three independent character sets was assessed. The first data set comprised the sequences of the 63 genes common to these viruses. The second set of characters was based on gene order, and phylogenies were inferred using both breakpoint distance analysis and a novel method developed here, termed neighbor pair analysis. The third set recorded gene content by scoring gene presence or absence in each genome. All three data sets yielded phylogenies supporting the separation of the Nucleopolyhedrovirus (NPV) and Granulovirus (GV) genera, the division of the NPVs into groups I and II, and species relationships within group I NPVs. Generation of phylogenies based on the combined sequences of all 63 shared genes proved to be the most effective approach to resolving the relationships among the group II NPVs and the GVs. The history of gene acquisitions and losses that have accompanied baculovirus diversification was visualized by mapping the gene content data onto the phylogenetic tree. This analysis highlighted the fluid nature of baculovirus genomes, with evidence of frequent genome rearrangements and multiple gene content changes during their evolution. Of more than 416 genes identified in the genomes analyzed, only 63 are present in all nine genomes, and 200 genes are found only in a single genome. Despite this fluidity, the whole genome-based methods we describe are sufficiently powerful to recover the underlying phylogeny of the viruses. PMID- 11483758 TI - Functional analysis of PA binding by influenza a virus PB1: effects on polymerase activity and viral infectivity. AB - Influenza A virus expresses three viral polymerase (P) subunits-PB1, PB2, and PA all of which are essential for RNA and viral replication. The functions of P proteins in transcription and replication have been partially elucidated, yet some of these functions seem to be dependent on the formation of a heterotrimer for optimal viral RNA transcription and replication. Although it is conceivable that heterotrimer subunit interactions may allow a more efficient catalysis, direct evidence of their essentiality for viral replication is lacking. Biochemical studies addressing the molecular anatomy of the P complexes have revealed direct interactions between PB1 and PB2 as well as between PB1 and PA. Previous studies have shown that the N-terminal 48 amino acids of PB1, termed domain alpha, contain the residues required for binding PA. We report here the refined mapping of the amino acid sequences within this small region of PB1 that are indispensable for binding PA by deletion mutagenesis of PB1 in a two-hybrid assay. Subsequently, we used site-directed mutagenesis to identify the critical amino acid residues of PB1 for interaction with PA in vivo. The first 12 amino acids of PB1 were found to constitute the core of the interaction interface, thus narrowing the previous boundaries of domain alpha. The role of the minimal PB1 domain alpha in influenza virus gene expression and genome replication was subsequently analyzed by evaluating the activity of a set of PB1 mutants in a model reporter minigenome system. A strong correlation was observed between a functional PA binding site on PB1 and P activity. Influenza viruses bearing mutant PB1 genes were recovered using a plasmid-based influenza virus reverse genetics system. Interestingly, mutations that rendered PB1 unable to bind PA were either nonviable or severely growth impaired. These data are consistent with an essential role for the N terminus of PB1 in binding PA, P activity, and virus growth. PMID- 11483759 TI - Simian immunodeficiency virus in which nef and U3 sequences do not overlap replicates efficiently in vitro and in vivo in rhesus macaques. AB - The nef genes of human immunodeficiency virus and simian immunodeficiency virus (SIV) overlap about 80% of the U3 region of the 3' long terminal repeat (LTR) and contain several essential cis-acting elements (here referred to as the TPI region): a T-rich region, the polypurine tract, and attachment (att) sequences required for integration. We inactivated the TPI region in the nef reading frame of the pathogenic SIVmac239 clone (239wt) by 13 silent point mutations. To restore viral infectivity, intact cis-regulatory elements were inserted just downstream of the mutated nef gene. The resulting SIV genome contains U3 regions that are 384 bp shorter than the 517-bp 239wt U3 region. Overall, elimination of the duplicated Nef coding sequences truncates the proviral genome by 350 bp. Nonetheless, it contains all known coding sequences and cis-acting elements. The TPI mutant virus expressed functional Nef and replicated like 239wt in all cell culture assays and in vivo in rhesus macaques. Notably, these SIVmac constructs allow us to study Nef function in the context of replication-competent viruses without the restrictions of overlapping LTR sequences and important cis-acting elements. The genomes of all known primate lentiviruses contain a large overlap between nef and the U3 region. We demonstrate that this conserved genomic organization is not obligatory for efficient viral replication and pathogenicity. PMID- 11483760 TI - Early polyadenylation signals of human papillomavirus type 31 negatively regulate capsid gene expression. AB - The L1 and L2 capsid genes of human papillomavirus type 31 (HPV-31) are expressed upon keratinocyte differentiation from a promoter located in the E7 open reading frame (ORF) of the early region. Late transcripts must therefore pass through and ignore the early polyadenylation sequences to use the downstream late AAUAAA element located at the end of the L1 ORF. To identify sequences which modulate downstream capsid gene expression, a variety of substitution mutations were introduced into the early polyadenylation signal and studied first in the context of polycistronic luciferase reporter constructs. Removal of the G/U-rich cleavage stimulation factor (CstF) binding sites and the degenerate cleavage and polyadenylation specificity factor binding sites, UAUAUA, had minimal effect on downstream expression as defined by luciferase activities. This is in contrast to the deletion of the HPV-31 early AAUAAA element, which resulted in a dramatic increase in downstream expression. Additional sequences within the first 800 bp of the L2 ORF were also found to negatively regulate capsid expression in luciferase assays. To determine how these mutations influence gene expression in the context of the complete HPV-31 genome, recombinant genomes were constructed that contained a substitution in the AAUAAA sequence, an inserted strong CstF binding site, an inserted simian virus 40 (SV40) late poly(A) signal, or a substitution of the 5'-most 800 nucleotides of the L2 ORF. Reductions in both transient and stable replication were observed with the recombinant genomes containing the strong CstF site or the late SV40 signal, suggesting that alterations in the strength of the upstream poly(A) signal influence expression of viral replication factors. Similarly, disruption of the L2 ORF resulted in a significant reduction in genome replication and an inability to be maintained stably. In contrast, genomes containing a substitution of the AAUAAA sequence had increased levels of transient and stable replication. Quantitation of late transcripts following keratinocyte differentiation in methylcellulose also showed a reduction in downstream capsid gene expression in lines containing genomes with the strong CstF site or the late SV40 signal mutations, while a significant increase in expression was detected in the lines with genomes lacking the AAUAAA sequence. These studies demonstrate that capsid gene expression in HPV-31 requires an inefficient early poly(A) signal which is defined primarily by the AAUAAA element as well as a major negative regulatory element located within the L2 ORF. PMID- 11483761 TI - Poliovirus 3A protein limits interleukin-6 (IL-6), IL-8, and beta interferon secretion during viral infection. AB - During viral infections, the host secretory pathway is crucial for both innate and acquired immune responses. For example, the export of most proinflammatory and antiviral cytokines, which recruit lymphocytes and initiate antiviral defenses, requires traffic through the host secretory pathway. To investigate potential effects of the known inhibition of cellular protein secretion during poliovirus infection on pathogenesis, cytokine secretion from cells infected with wild-type virus and with 3A-2, a mutant virus carrying an insertion in viral protein 3A which renders the virus defective in the inhibition of protein secretion, was tested. We show here that cells infected with 3A-2 mutant virus secrete greater amounts of cytokines interleukin-6 (IL-6), IL-8, and beta interferon than cells infected with wild-type poliovirus. Increased cytokine secretion from the mutant-infected cells can be attributed to the reduced inhibition of host protein secretion, because no significant differences between 3A-2- and wild-type-infected cells were observed in the inhibition of viral growth, host cell translation, or the ability of wild-type- or 3A-2-infected cells to support the transcriptional induction of beta interferon mRNA. We surmise that the wild-type function of 3A in inhibiting ER-to-Golgi traffic is not required for viral replication in tissue culture but, by altering the amount of secreted cytokines, could have substantial effects on pathogenesis within an infected host. The global inhibition of protein secretion by poliovirus may reflect a general mechanism by which pathogens that do not require a functional protein secretory apparatus can reduce the native immune response and inflammation associated with infection. PMID- 11483762 TI - The 2.2-kilobase latency-associated transcript of herpes simplex virus type 2 does not modulate viral replication, reactivation, or establishment of latency in transgenic mice. AB - To better understand the mechanisms responsible for the observed effects of deletions in the promoter region of the latency-associated transcript (LAT) gene in impairing herpes simplex virus (HSV) reactivation, we generated mice transgenic for a 5.5-kb HSV type 2 (HSV-2) genomic fragment spanning the major LAT, along with the LAT promoter and flanking regions, in the C57BL/6 background. The mice expressed abundant 2.2-kb major LATs in trigeminal ganglia (TG) and other tissues. The effects of the transgene on HSV-2 infection, latency, and reactivation were assessed. When infected with wild-type (WT) HSV-2 or its LAT promoter deletion (LAT(-)) mutant, primary lung fibroblast lines established from normal C57BL/6 and transgenic mice supported virus growth equally well. The replication of these viruses in the mouse eye and their spread to TG and brains were similar. The quantities of latent viral DNA in TG of transgenic and normal mice, as determined by real-time PCR, were comparable. UV light-induced reactivation of the LAT(-) mutant from transgenic mice (0 to 7%) was no more frequent than that from normal mice (0 to 14%), while WT virus was reactivated from 13 to 54% of normal mice and 22 to 54% of transgenic mice. The cumulative data indicate that, when expressed transgenically, the HSV-2 major LAT cannot influence HSV-2 infection or latency and cannot complement the defect in reactivation of the LAT(-) mutant. These results imply that the phenotype of reduced reactivation associated with the LAT(-) mutant is related to a function encoded in the LAT promoter but not to the major LAT itself. PMID- 11483764 TI - Expression of immunoregulatory cytokines by recombinant coxsackievirus B3 variants confers protection against virus-caused myocarditis. AB - Clinical and laboratory investigations have demonstrated the involvement of viruses and bacteria as potential causative agents in cardiovascular disease and have specifically found coxsackievirus B3 (CVB3) to be a leading cause. Experimental data indicate that cytokines are involved in controlling CVB3 replication. Therefore, recombinant CVB3 (CVB3rec) variants expressing the T helper-1 (T(H)1)-specific gamma interferon (IFN-gamma) or the T(H)2-specific interleukin-10 (IL-10) as well as the control virus CVB3(muIL-10), which produce only biologically inactive IL-10, were established. Coding regions of murine cytokines were cloned into the 5' end of the CVB3 wild type (CVB3wt) open reading frame and were supplied with an artificial viral 3Cpro-specific Q-G cleavage site. Correct processing releases active cytokines, and the concentration of IFN gamma and IL-10 was analyzed by enzyme-linked immunosorbent assay and bioassays. In mice, CVB3wt was detectable in pancreas and heart tissue, causing massive destruction of the exocrine pancreas as well as myocardial inflammation and heart cell lysis. Most of the CVB3wt-infected mice revealed virus-associated symptoms, and some died within 28 days postinfection. In contrast, CVB3rec variants were present only in the pancreas of infected mice, causing local inflammation with subsequent healing. Four weeks after the first infection, surviving mice were challenged with the lethal CVB3H3 variant, causing casualties in the CVB3wt- and CVB3(muIL-10)-infected groups, whereas almost none of the CVB3(IFN-gamma)- and CVB3(IL-10)-infected mice died and no pathological disorders were detectable. This study demonstrates that expression of immunoregulatory cytokines during CVB3 replication simultaneously protects mice against a lethal disease and prevents virus-caused tissue destruction. PMID- 11483763 TI - Targeted disruption of the Ceacam1 (MHVR) gene leads to reduced susceptibility of mice to mouse hepatitis virus infection. AB - The CEACAM1 glycoproteins (formerly called biliary glycoproteins; BGP, C-CAM, CD66a, or MHVR) are members of the carcinoembryonic antigen family of cell adhesion molecules. In the mouse, splice variants of CEACAM1 have either two or four immunoglobulin (Ig) domains linked through a transmembrane domain to either a short or a long cytoplasmic tail. CEACAM1 has cell adhesion activity and acts as a signaling molecule, and long-tail isoforms inhibit the growth of colon and prostate tumor cells in rodents. CEACAM1 isoforms serve as receptors for several viral and bacterial pathogens, including the murine coronavirus mouse hepatitis virus (MHV) and Haemophilus influenzae, Neisseria gonorrhoeae, and Neisseria meningitidis in humans. To elucidate the mechanisms responsible for the many biological activities of CEACAM1, we modified the expression of the mouse Ceacam1 gene in vivo. Manipulation of the Ceacam1 gene in mouse embryonic stem cells that contained the Ceacam1a allele yielded a partial knockout. We obtained one line of mice in which the insert in the Ceacam1a gene had sustained a recombination event. This resulted in the markedly reduced expression of the two CEACAM1a isoforms with four Ig domains, whereas the expression of the two isoforms with two Ig domains was doubled relative to that in wild-type BALB/c (+/+) mice. Homozygous (p/p) Ceacam1a-targeted mice (Ceacam1aDelta4D) had no gross tissue abnormalities and were viable and fertile; however, they were more resistant to MHV A59 infection and death than normal (+/+) mice. Following intranasal inoculation with MHV A59, p/p mice developed markedly fewer and smaller lesions in the liver than +/+ or heterozygous (+/p) mice. The titers of virus produced in the livers were 50- to 100-fold lower in p/p mice than in +/p or +/+ mice. p/p mice survived a dose 100-fold higher than the lethal dose of virus for +/+ mice. +/p mice were intermediate between +/+ and p/p mice in susceptibility to liver damage, virus growth in liver, and susceptibility to killing by MHV. Ceacam1a targeted mice provide a new model to study the effects of modulation of receptor expression on susceptibility to MHV infection in vivo. PMID- 11483765 TI - Interleukin-8 stimulates human immunodeficiency virus type 1 replication and is a potential new target for antiretroviral therapy. AB - Production of the C-X-C chemokines interleukin-8 (IL-8) and growth-regulated oncogene alpha (GRO-alpha) in macrophages is stimulated by exposure to human immunodeficiency virus type 1 (HIV-1). We have demonstrated previously that GRO alpha then stimulates HIV-1 replication in both T lymphocytes and macrophages. Here we demonstrate that IL-8 also stimulates HIV-1 replication in macrophages and T lymphocytes. We further show that increased levels of IL-8 are present in the lymphoid tissue of patients with AIDS. In addition, we demonstrate that compounds which inhibit the actions of IL-8 and GRO-alpha via their receptors, CXCR1 and CXCR2, also inhibit HIV-1 replication in both T lymphocytes and macrophages, indicating potential therapeutic uses for these compounds in HIV-1 infection and AIDS. PMID- 11483766 TI - Inhibition of human immunodeficiency virus type 1 Rev function by a dominant negative mutant of Sam68 through sequestration of unspliced RNA at perinuclear bundles. AB - Human immunodeficiency virus (HIV) type 1 encodes an essential protein, Rev, which functions to transport unspliced and singly spliced viral transcripts from the nucleus to the cytoplasm to allow expression of the viral structural proteins. It has previously been reported that Sam68 synergistically stimulates Rev activity (T. Reddy et al., Nat. Med. 5:635-642, 1999). Here we report that the Sam68-like mammalian proteins SLM1 and SLM2 also stimulate Rev activity. Their stimulation ability cannot be attributed to a shuttling property, since Sam68, SLM1, and SLM2 do not display significant shuttling activity alone or in the presence of Rev. In addition, Sam68, SLM1, and SLM2 do not affect the equilibrium between unspliced and completely spliced HIV RNA. The C-terminally truncated Sam68 mutant (Sam68DeltaC) previously observed to inhibit the Sam68 mediated stimulation of Rev activity (Reddy et al., 1999) also inhibits SLM1- and SLM2-mediated stimulation of Rev activity. This suggests that the mechanism by which Sam68, SLM1, and SLM2 stimulate Rev activity may be common. Sam68DeltaC does not inhibit Rev activity by inhibiting Rev from shuttling between the nucleus and cytoplasm. Inhibition by Sam68DeltaC is a consequence of its mislocalization to the cytoplasm, as evidenced by the fact that addition of an exogenous nuclear localization signal to Sam68DeltaC restores nuclear localization and stimulation of Rev activity. We demonstrate that Sam68DeltaC causes perinuclear accumulation of unspliced HIV env RNA and propose that Sam68DeltaC inhibits Rev activity by sequestering Rev-responsive RNA away from the translation apparatus. PMID- 11483767 TI - Alpha/beta interferon promotes transcription and inhibits replication of borna disease virus in persistently infected cells. AB - Borna disease virus (BDV) is a noncytolytic RNA virus that can replicate in the central nervous system (CNS) of mice. This study shows that BDV multiplication was efficiently blocked in transgenic mice that express mouse alpha-1 interferon (IFN-alpha1) in astrocytes. To investigate whether endogenous virus-induced IFN might similarly restrict BDV, we used IFNAR(0/0) mice, which lack a functional alpha/beta IFN (IFN-alpha/beta) receptor. As would be expected if virus-induced IFN were important to control BDV infection, we found that cultured embryo cells of IFNAR(0/0) mice supported viral multiplication, whereas cells from wild-type mice did not. Unexpectedly, however, BDV spread through the CNSs of IFNAR(0/0) and wild-type mice with similar kinetics, suggesting that activation of endogenous IFN-alpha/beta genes in BDV-infected brains was too weak or occurred too late to be effective. Surprisingly, Northern blot analysis showed that the levels of the most abundant viral mRNAs in the brains of persistently infected IFNAR(0/0) mice were about 20-fold lower than those in wild-type mice. In contrast, genomic viral RNA was produced in about a 10-fold excess in the brains of IFNAR(0/0) mice. Human IFN-alpha2 similarly enhanced transcription and simultaneously repressed replication of the BDV genome in persistently infected Vero cells. Thus, in persistently infected neurons and cultured cells, IFN alpha/beta appears to freeze the BDV polymerase in the transcriptional mode, resulting in enhanced viral mRNA synthesis and suppressing viral genome replication. PMID- 11483768 TI - Mutational analysis of the repeated open reading frames, ORFs 63 and 70 and ORFs 64 and 69, of varicella-zoster virus. AB - Varicella-zoster virus (VZV) open reading frame 63 (ORF63), located between nucleotides 110581 and 111417 in the internal repeat region, encodes a nuclear phosphoprotein which is homologous to herpes simplex virus type 1 (HSV-1) ICP22 and is duplicated in the terminal repeat region as ORF70 (nucleotides 118480 to 119316). We evaluated the role of ORFs 63 and 70 in VZV replication, using recombinant VZV cosmids and PCR-based mutagenesis to make single and dual deletions of these ORFs. VZV was recovered within 8 to 10 days when cosmids with single deletions were transfected into melanoma cells along with the three intact VZV cosmids. In contrast, VZV was not detected in transfections carried out with a dual deletion cosmid. Infectious virus was recovered when ORF63 was cloned into a nonnative AvrII site in this cosmid, confirming that failure to generate virus was due to the dual ORF63/70 deletion and that replication required at least one gene copy. This requirement may be related to our observation that ORF63 interacts directly with ORF62, the major immediate-early transactivating protein of VZV. ORF64 is located within the inverted repeat region between nucleotides 111565 and 112107; it has some homology to the HSV-1 Us10 gene and is duplicated as ORF69 (nucleotides 117790 to 118332). ORF64 and ORF69 were deleted individually or simultaneously using the VZV cosmid system. Single deletions of ORF64 or ORF69 yielded viral plaques with the same kinetics and morphology as viruses generated with the parental cosmids. The dual deletion of ORF64 and ORF69 was associated with an abnormal plaque phenotype characterized by very large, multinucleated syncytia. Finally, all of the deletion mutants that yielded recombinants retained infectivity for human T cells in vitro and replicated efficiently in human skin in the SCIDhu mouse model of VZV pathogenesis. PMID- 11483769 TI - Nonenveloped nucleocapsids of hepatitis C virus in the serum of infected patients. AB - One of the characteristics of hepatitis C virus (HCV) is the high incidence of persistent infection. HCV core protein, in addition to forming the viral nucleocapsid, has multiple regulatory functions in host-cell transcription, apoptosis, cell transformation, and lipid metabolism and may play a role in suppressing host immune response. This protein is thought to be present in the bloodstream of the infected host as the nucleocapsid of infectious, enveloped virions. This study provides evidence that viral particles with the physicochemical, morphological, and antigenic properties of nonenveloped HCV nucleocapsids are present in the plasma of HCV-infected individuals. These particles have a buoyant density of 1.32 to 1.34 g/ml in CsCl, are heterogeneous in size (with predominance of particles 38 to 43 or 54 to 62 nm in diameter on electron microscopy), and express on their surface epitopes located in amino acids 24 to 68 of the core protein. Similar nucleocapsid-like particles are also produced in insect cells infected with recombinant baculovirus bearing cDNA for structural HCV proteins. HCV core particles isolated from plasma were used to generate anti-core monoclonal antibodies (MAbs). These MAbs stained HCV core in the cytoplasm of hepatocytes from experimentally infected chimpanzees in the acute phase of the infection. These chimpanzees had concomitantly HCV core antigen in serum. These findings suggest that overproduction of nonenveloped nucleocapsids and their release into the bloodstream are properties of HCV morphogenesis. The presence of circulating cores in serum and accumulation of the core protein in liver cells during the early phase of infection may contribute to the persistence of HCV and its many immunopathological effects in the infected host. PMID- 11483770 TI - Bovine herpesvirus 1 tegument protein VP22 interacts with histones, and the carboxyl terminus of VP22 is required for nuclear localization. AB - The bovine herpesvirus 1 (BHV-1) UL49 gene encodes a viral tegument protein termed VP22. UL49 homologs are conserved among alphaherpesviruses. Interestingly, the BHV-1 VP22 deletion mutant virus is asymptomatic and avirulent in infected cattle but produces only a slight reduction in titer in vitro. Attenuation of the BHV-1 VP22 deletion mutant virus in vivo suggests that VP22 plays a functional role in BHV-1 replication. In herpes simplex virus type 1, the VP22 homolog was previously shown to interact with another tegument protein,VP16, the alpha transinducing factor in vitro. In this report, we show that (i) the nuclear targeting of VP22 is independent of other viral factors, (ii) the carboxyl terminus of VP22 is required for its nuclear localization, (iii) VP22 associates with histones and nucleosomes, (iv) an antihistone monoclonal antibody cross reacts with VP22, and (v) acetylation of histone H4 is decreased in VP22 expressing cells as well as virus-infected cells. Our data suggest that VP22 may have a modulatory function during BHV-1 infection. PMID- 11483771 TI - Tick-borne Langat/mosquito-borne dengue flavivirus chimera, a candidate live attenuated vaccine for protection against disease caused by members of the tick borne encephalitis virus complex: evaluation in rhesus monkeys and in mosquitoes. AB - Langat virus (LGT), strain TP21, a naturally avirulent tick-borne flavivirus, was used to construct a chimeric candidate virus vaccine which contained LGT genes for premembrane (preM) and envelope (E) glycoprotein and all other sequences derived from dengue type 4 virus (DEN4). The live virus vaccine was developed to provide resistance to the highly virulent, closely related tick-borne flaviviruses that share protective E epitopes among themselves and with LGT. Toward that end the chimera, initially recovered in mosquito cells, was adapted to grow to high titer in qualified simian Vero cells. When inoculated intraperitoneally (i.p.), the Vero cell-adapted LGT TP21/DEN4 chimera remained completely attenuated for SCID mice. Significantly, the chimera protected immunocompetent mice against the most virulent tick-borne encephalitis virus (TBEV). Subsequently, rhesus monkeys were immunized in groups of 4 with 10(5) or 10(7) PFU of LGT strain TP21, with 10(5) PFU of DEN4, or with 10(3), 10(5), or 10(7) PFU of the chimera. Each of the monkeys inoculated with DEN4 or LGT TP21 became viremic, and the duration of viremia ranged from 1 to 5 days. In contrast, viremia was detected in only 1 of 12 monkeys inoculated with the LGT TP21/DEN4 chimera; in this instance the level of viremia was at the limit of detection. All monkeys immunized with the chimera or LGT TP21 virus developed a moderate to high level of neutralizing antibodies against LGT TP21 as well as TBEV and were completely protected against subsequent LGT TP21 challenge, whereas monkeys previously immunized with DEN4 virus became viremic when challenged with LGT TP21. These observations suggest that the chimera is attenuated, immunogenic, and able to induce a protective immune response. Furthermore, passive transfer of serum from monkeys immunized with chimera conferred significant protection to mice subsequently challenged with 100 i.p. 50% lethal doses of the highly virulent TBEV. The issue of transmissibility of the chimera by mosquitoes was addressed by inoculating a nonhematophagous mosquito, Toxorhynchites splendens, intrathoracically with the chimera or its DEN4 or LGT parent. Neither the LGT TP21/DEN4 vaccine candidate nor the wild-type LGT TP21 virus was able to infect this mosquito species, which is highly permissive for dengue viruses. Certain properties of the chimera, notably its attenuation for monkeys, its immunogenicity, and its failure to infect a highly permissive mosquito host, make it a promising vaccine candidate for use in immunization against severe disease caused by many tick-borne flaviviruses. PMID- 11483772 TI - Morbillivirus infection of the mouse central nervous system induces region specific upregulation of MMPs and TIMPs correlated to inflammatory cytokine expression. AB - Viral infection of the central nervous system (CNS) can result in perturbation of cell-to-cell communication involving the extracellular matrix (ECM). ECM integrity is maintained by a dynamic balance between the synthesis and proteolysis of its components, mainly as a result of the action of matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs). An MMP/TIMP imbalance may be critical in triggering neurological disorders, in particular in virally induced neural disorders. In the present study, a mouse model of brain infection using a neurotropic strain of canine distemper virus (CDV) was used to study the effect of CNS infection on the MMP/TIMP balance and cytokine expression. CDV replicates almost exclusively in neurons and has a unique pattern of expression (cortex, hypothalamus, monoaminergic nuclei, hippocampus, and spinal cord). Here we show that although several mouse brain structures were infected, they exhibited a differential pattern in terms of MMP, TIMP, and cytokine expression, exemplified by (i) a large increase in pro-MMP9 levels, in particular in the hippocampus, which occurred mainly in neurons and was associated with in situ gelatinolytic activity, (ii) specific and significant upregulation of MT1-MMP mRNA expression in the cortex and hypothalamus, (iii) an MMP/TIMP imbalance, suggested by the upregulation of TIMP-1 mRNA in the cortex, hippocampus, and hypothalamus and of TIMP-3 mRNA in the cortex, and (iv) a concomitant region-specific large increase in expression of Th1-like cytokines, such as gamma interferon, tumor necrosis factor alpha, and interleukin 6 (IL-6), contrasting with weaker induction of Th2 like cytokines, such as IL-4 and IL-10. These data indicate that an MMP/TIMP imbalance in specific brain structures, which is tightly associated with a local inflammatory process as shown by the presence of immune infiltrating cells, differentially impairs CNS integrity and may contribute to the multiplicity of late neurological disorders observed in this viral mouse model. PMID- 11483773 TI - Latent antigen vaccination in a model gammaherpesvirus infection. AB - Vaccines that can reduce the load of latent gammaherpesvirus infections are eagerly sought. One attractive strategy is vaccination against latency-associated proteins, which may increase the efficiency with which T cells recognize and eliminate latently infected cells. However, due to the lack of tractable animal model systems, the effect of latent-antigen vaccination on gammaherpesvirus latency is not known. Here we use the murine gammaherpesvirus model to investigate the impact of vaccination with the latency-associated M2 antigen. As expected, vaccination had no effect on the acute lung infection. However, there was a significant reduction in the load of latently infected cells in the initial stages of the latent infection, when M2 is expressed. These data show for the first time that latent-antigen vaccination can reduce the level of latency in vivo and suggest that vaccination strategies involving other latent antigens may ultimately be successfully used to reduce the long-term latent infection. PMID- 11483774 TI - Structure-based mutational analysis of the hepatitis C virus NS3 helicase. AB - The carboxyl terminus of the hepatitis C virus (HCV) nonstructural protein 3 (NS3) possesses ATP-dependent RNA helicase activity. Based on the conserved sequence motifs and the crystal structures of the helicase domain, 17 mutants of the HCV NS3 helicase were generated. The ATP hydrolysis, RNA binding, and RNA unwinding activities of the mutant proteins were examined in vitro to determine the functional role of the mutated residues. The data revealed that Lys-210 in the Walker A motif and Asp-290, Glu-291, and His-293 in the Walker B motif were crucial to ATPase activity and that Thr-322 and Thr-324 in motif III and Arg-461 in motif VI significantly influenced ATPase activity. When the pairing between His-293 and Gln-460, referred to as gatekeepers, was replaced with the Asp 293/His-460 pair, which makes the NS3 helicase more like the DEAD helicase subgroup, ATPase activity was not restored. It thus indicated that the whole microenvironment surrounding the gatekeepers, rather than the residues per se, was important to the enzymatic activities. Arg-461 and Trp-501 are important residues for RNA binding, while Val-432 may only play a coadjutant role. The data demonstrated that RNA helicase activity was possibly abolished by the loss of ATPase activity or by reduced RNA binding activity. Nevertheless, a low threshold level of ATPase activity was found sufficient for helicase activity. Results in this study provide a valuable reference for efforts under way to develop anti-HCV therapeutic drugs targeting NS3. PMID- 11483775 TI - Occurrence of genetic drift and founder effect during quasispecies evolution of the VP2 and NS3/NS3A genes of bluetongue virus upon passage between sheep, cattle, and Culicoides sonorensis. AB - Bluetongue virus (BTV) is the cause of an insect-transmitted virus infection of ruminants that occurs throughout much of the world. Individual gene segments differ between field strains of BTV; thus, we hypothesized that key viral genes undergo genetic drift during alternating passage of BTV in its ruminant and insect hosts. To test this hypothesis, variation in the consensus sequence and quasispecies heterogeneity of the VP2 and NS3/NS3A genes of a plaque-purified strain of BTV serotype 10 was determined during alternating infection of vector Culicoides sonorensis and a sheep and calf. Consensus sequences were determined after reverse transcriptase-nested PCR amplification of viral RNA directly from ruminant blood and homogenized insects, and quasispecies heterogeneity was determined by the sequencing of clones derived from directly amplified viral RNA. Comparison of these sequences to those of the original BTV inoculum used to initiate the cycle of BTV infection demonstrated, for the first time, that individual BTV gene segments evolve independently of one another by genetic drift in a host-specific fashion, generating quasispecies populations in both ruminant and insect hosts. Furthermore, a unique viral variant was randomly ingested by C. sonorensis insects that fed on a sheep with low-titer viremia, thereby fixing a novel genotype by founder effect. Thus, we conclude that genetic drift and founder effect contribute to diversification of individual gene segments of field strains of BTV. PMID- 11483776 TI - Noncytolytic inhibition of X4 virus by bulk CD8(+) cells from human immunodeficiency virus type 1 (HIV-1)-infected persons and HIV-1-specific cytotoxic T lymphocytes is not mediated by beta-chemokines. AB - Human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) mediate immunologic selection pressure by both cytolytic and noncytolytic mechanisms. Non cytolytic mechanisms include the release of beta-chemokines blocking entry of R5 HIV-1 strains. In addition, CD8(+) cells inhibit X4 virus isolates via release of as yet poorly characterized soluble factors. To further characterize these factors, we performed detailed analysis of CTL as well as bulk CD8(+) T lymphocytes from six HIV-1-infected individuals and from six HIV-1-seronegative individuals. Kinetic studies revealed that secreted suppressive activities of HIV 1-specific CTL and bulk CD8(+) T lymphocytes from all HIV-1-infected persons are significantly higher than that of supernatants from seronegative controls. The suppressive activity could be blocked by monensin and brefeldin A, was heat labile, and appeared in a pattern different from that of secretion of chemokines (MDC, I-309, MIP-1alpha, MIP-1beta, and RANTES), cytokines (gamma interferon, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor), and interleukins (interleukin-13 and interleukin-16). This suppression activity was characterized by molecular size exclusion centrifugation and involves a suppressive activity of >50 kDa which could be bound to heparin and a nonbinding inhibitory activity of <50 kDa. Our data provide a functional link between CD8(+) cells and CTL in the noncytolytic inhibition of HIV-1 and suggest that suppression of X4 virus is mediated through proteins. The sizes of the proteins, their affinity for heparin, and the pattern of release indicate that these molecules are not chemokines. PMID- 11483777 TI - Human T-cell leukemia virus type 1 receptor expression among syncytium-resistant cell lines revealed by a novel surface glycoprotein-immunoadhesin. AB - The envelope glycoproteins of human T-cell leukemia virus type 1 (HTLV-1) perform functions that are crucial for virus entry into cells. The surface glycoprotein (SU) is responsible for viral recognition of, and binding to, target cells through its interaction with an unknown cell surface receptor. To facilitate molecular analysis of the receptor-binding properties of SU and to characterize the cellular receptor employed by HTLV-1, we have expressed a recombinant SU fused to the Fc domain of human immunoglobulin G. Here, we demonstrate that this novel SU-immunoadhesin retains both the biochemical properties of Fc and the receptor-binding specificity of the HTLV-1 SU. We use this SU-immunoadhesin to demonstrate, by direct cell surface binding assays, that the receptor used by HTLV-1 has been conserved through vertebrate evolution. Moreover, using murine human somatic cell hybrids we provide data that do not support the previously assigned location for the HTLV-1 receptor on human chromosome 17. Most importantly, we show that many cell lines that are resistant to HTLV-1 envelope mediated infection and syncytium formation express functional receptors that are recognized by the HTLV-1 SU. Based on our results, we suggest that for some HTLV 1-resistant cell lines the block to viral entry occurs at a late post-receptor binding step of the entry process. Our findings will be of value in developing new strategies to identify the cellular receptor used by HTLV-1. PMID- 11483778 TI - In vivo generation and characterization of a soluble form of the Semliki forest virus fusion protein. AB - During infection of host cells, a number of enveloped animal viruses are known to produce soluble forms of viral membrane glycoproteins lacking the transmembrane domain. The roles of such soluble glycoproteins in viral life cycles are incompletely understood, but in several cases they are believed to modulate host immune response and viral pathogenesis. Semliki Forest virus (SFV) is an enveloped alphavirus that infects cells through low-pH-dependent fusion and buds from the plasma membrane. Fusion is mediated by the E1 subunit of the SFV spike protein. Previous studies described the in vivo generation of E1s, a truncated soluble form of E1, under conditions in which budding is inhibited in mammalian host cells. We have here examined the properties of E1s generation and the biological activity of E1s. E1s cleavage required spike protein transport out of the endoplasmic reticulum and was independent of virus infection. Cell surface E1 efficiently acted as a precursor for E1s. E1s generation was strongly pH dependent in BHK cells, with optimal cleavage at a pH of < or =7.0, conditions that inhibited the budding of SFV but not the budding of the rhabdovirus vesicular stomatitis virus. The pH dependence of E1s production and SFV budding was unaffected by the stability of the spike protein dimer but was a function of the host cell. Similar to the intact virus and in vitro-generated E1 ectodomain, treatment of E1s at low pH in the presence of target membranes triggered specific acid-dependent conformational changes. Thus, under a variety of conditions, SFV infected cells can produce a soluble form of E1 that is biologically active. PMID- 11483779 TI - Antibody protects macaques against vaginal challenge with a pathogenic R5 simian/human immunodeficiency virus at serum levels giving complete neutralization in vitro. AB - A major unknown in human immunodeficiency virus (HIV-1) vaccine design is the efficacy of antibodies in preventing mucosal transmission of R5 viruses. These viruses, which use CCR5 as a coreceptor, appear to have a selective advantage in transmission of HIV-1 in humans. Hence R5 viruses predominate during primary infection and persist throughout the course of disease in most infected people. Vaginal challenge of macaques with chimeric simian/human immunodeficiency viruses (SHIV) is perhaps one of the best available animal models for human HIV-1 infection. Passive transfer studies are widely used to establish the conditions for antibody protection against viral challenge. Here we show that passive intravenous transfer of the human neutralizing monoclonal antibody b12 provides dose-dependent protection to macaques vaginally challenged with the R5 virus SHIV(162P4). Four of four monkeys given 25 mg of b12 per kg of body weight 6 h prior to challenge showed no evidence of viral infection (sterile protection). Two of four monkeys given 5 mg of b12/kg were similarly protected, whereas the other two showed significantly reduced and delayed plasma viremia compared to control animals. In contrast, all four monkeys treated with a dose of 1 mg/kg became infected with viremia levels close to those for control animals. Antibody b12 serum concentrations at the time of virus challenge corresponded to approximately 400 (25 mg/kg), 80 (5 mg/kg), and 16 (1 mg/kg) times the in vitro (90%) neutralization titers. Therefore, complete protection against mucosal challenge with an R5 SHIV required essentially complete neutralization of the infecting virus. This suggests that a vaccine based on antibody alone would need to sustain serum neutralizing antibody titers (90%) of the order of 1:400 to achieve sterile protection but that lower titers, around 1:100, could provide a significant benefit. The significance of such substerilizing neutralizing antibody titers in the context of a potent cellular immune response is an important area for further study. PMID- 11483780 TI - Nuclear export of human immunodeficiency virus type 1 Vpr is not required for virion packaging. AB - The human immunodeficiency virus type 1 Vpr protein is both packaged into virions and efficiently localized to the nucleus. In this report, we show that a significant fraction of Vpr also accumulates in the cytoplasm of virus-producing cells. Although Vpr shuttles between the nucleus and the cytoplasm, studies with an export-deficient Vpr mutant reveal that nuclear export is not required for virion incorporation. PMID- 11483781 TI - Natural history of neonatal herpes simplex virus infections in the acyclovir era. AB - OBJECTIVE: During the 2 decades in which effective antiviral therapies have been available for neonatal herpes simplex virus (HSV) disease, changes have been documented not only in the outcomes of infected infants, but also in the natural history of the disease itself. Numerous studies previously have reported that early institution of antiviral therapy is beneficial to the outcome of the disease. The objective of this study was to provide an update of neonatal HSV disease to identify means by which future improvements in the management of HSV infected neonates can be made. DESIGN/METHODS: Neonates enrolled in 2 studies of parenteral acyclovir for the treatment of neonatal HSV disease provided the data source. The National Institute of Allergy and Infectious Diseases Collaborative Antiviral Study Group conducted the studies between 1981 and 1997. A total of 186 patients are summarized, all of whom were treated with acyclovir. Demographic and clinical characteristics of these patients are reported. RESULTS: Comparisons between patients treated in the periods between 1981-1988 and 1989-1997 according to extent of disease revealed that the mean time between the onset of disease symptoms and initiation of therapy has not changed significantly from the early 1980s to the late 1990s. Of all patients evaluated, 40% had fetal scalp monitors during the delivery process. A significant minority of patients did not have skin vesicles at the time of their presentation and did not develop them during the acute HSV disease (39% of patients with disseminated disease; 32% of patients with central nervous system [CNS] disease; and 17% of patients with skin, eye, and/or mouth disease). Among patients with CNS disease, mortality was associated with prematurity. Among patients with disseminated HSV disease treated with acyclovir at 30 mg/kg/d, mortality was associated with aspartate transaminase elevations of >/=10 times the upper limit of normal at the time of initiation of acyclovir therapy. Mortality was also associated with lethargy at initiation of antiviral therapy for patients with disseminated disease. Patients' morbidity status was associated with the extent of disease (skin, eye, and/or mouth disease vs CNS vs disseminated). For those patients with CNS disease, morbidity was also associated with seizures at initiation of antiviral therapy. CONCLUSION: Data presented in the current comparison of neonatal HSV disease over the 2 periods (1981-1988 vs 1989-1997) demonstrate that no progress has been made in decreasing the interval between onset of HSV symptoms and initiation of antiviral therapy. Additional strides in the improvement of disease outcome may occur only if the interval between onset of symptoms and initiation of therapy is shortened. The means by which this will be accomplished lie in increased consideration of neonatal HSV infections in acutely ill infants. Specific data and recommendations to facilitate this goal are contained within. PMID- 11483782 TI - Safety and efficacy of high-dose intravenous acyclovir in the management of neonatal herpes simplex virus infections. AB - OBJECTIVE: The objective of this investigation was to establish the safety of high-dose (HD) acyclovir for the treatment of neonatal herpes simplex virus (HSV) disease. In addition, an estimate of therapeutic efficacy was sought, both with respect to mortality and to morbidity. Virologic efficacy of HD acyclovir was also assessed. PARTICIPANTS: Infants who were 5 million episodes of acute otitis media (AOM) accounted for $3 billion in health care expenditures. OBJECTIVES: To synthesize the literature on the natural history of AOM, the effectiveness of antibiotic treatment in uncomplicated AOM, and the relative effectiveness of specific antibiotic regimens. DATA SOURCES: Seven electronic databases for articles published between 1966 and March 1999 and reference lists in proceedings, published articles, reports, and guidelines. STUDY SELECTION: Two physicians independently assessed each article. Studies addressing AOM in children 4 weeks to 18 years old were included; those addressing children with immunodeficiencies or craniofacial abnormalities were excluded. Randomized, controlled trials (RCTs) were used to assess antibiotic effectiveness, and RCTs and cohort studies were used to assess the natural history of AOM. Among the 3491 citations identified, 80 (2.3%) met our inclusion criteria. DATA EXTRACTION: Two physicians independently abstracted data and assessed the quality of studies using a validated scale for RCTs and 8 quality components for cohort studies. DATA SYNTHESIS: Random-effects estimates of pooled absolute rate differences of outcomes were derived, and heterogeneity of both the rates and rate differences was assessed. Children with AOM not treated with antibiotics experienced a 1- to 7-day clinical failure rate of 19% (95% confidence interval: 0.10-0.28) and few suppurative complications. When patients were treated with amoxicillin, the 2- to 7-day clinical failure rate was reduced to 7%, a 12% (95% confidence interval: 0.04-0.20) reduction. Adverse effects, primarily gastrointestinal, were more common among children on cefixime than among those on ampicillin or amoxicillin. They were also more common among children on amoxicillin-clavulanate than among those on azithromycin. CONCLUSIONS: The majority of uncomplicated cases of AOM resolve spontaneously without apparent suppurative complications. Ampicillin or amoxicillin confers a limited therapeutic benefit. There is no evidence to support any particular antibiotic regimens as more effective at relieving symptoms. Certain antibiotics are more likely than others to cause diarrhea and other adverse events. PMID- 11483784 TI - Evidence assessment of management of acute otitis media: II. Research gaps and priorities for future research. AB - OBJECTIVES: To report research gaps and priorities of future research identified during an evidence assessment process on the management of acute otitis media (AOM). METHODS: A conceptual framework for management of AOM was developed to guide the evidence assessment. An 11-member technical expert panel guided the selection of key questions, prioritization of influencing factors, development of scope, definition of AOM, and search strategy through polling processes and conference calls. Quality of clinical trials was evaluated using established scales. Outcome measures were abstracted from each study. RESULTS: A total of 3461 titles and abstracts were screened, and 760 full-length articles were reviewed. Of the 760 articles, 80 studies addressed the key questions. In defining AOM, 42 (52.5%) of the 80 studies included the middle-ear effusion component, only 2 (2.5%) included the rapid onset component, and 26 (32.5%) included the signs/symptoms of inflammation component. None of the 80 studies used all 3 components. Of the 74 controlled trials, 39 (53%) were of acceptable quality (Jadad score of 3 or higher). The technical experts did not agree in the ranking of the importance of the 41 influencing factors (Kendall's coefficient of concordance was 0.0022). Another poll also indicated diverse opinions of the experts on the importance of 7 key questions derived from the conceptual framework (Kendall coefficient of concordance is 0.21). Furthermore, our review found that the type and definition of outcome measure varied. CONCLUSIONS: Despite the large body of literature on AOM, its quality is uneven and its findings are not generalizable. Future research should try to answer all key questions and investigate all risk factors in well-designed, scientific studies. PMID- 11483785 TI - Children with headache suspected of having a brain tumor: a cost-effectiveness analysis of diagnostic strategies. AB - OBJECTIVE: To assess the clinical and economic consequences of 3 diagnostic strategies-magnetic resonance imaging (MRI), computed tomography followed by MRI for positive results (CT-MRI), and no neuroimaging with close clinical follow-up in the evaluation of children with headache suspected of having a brain tumor. Three risk groups based on clinical variables were evaluated. MATERIALS AND METHODS: A decision-analytic Markov model and cost-effectiveness analysis was performed incorporating the risk group prior probability, MRI and CT sensitivity and specificity, tumor survival, progression rates, and cost per strategy. Outcomes were based on quality-adjusted life year (QALY) gained and incremental cost per QALY gained. RESULTS: For low-risk children with chronic nonmigraine headaches of >6 months' duration as the sole symptom (prior probability of brain tumor 0.01%), no neuroimaging with close clinical follow-up was less costly and more effective than the 2 neuroimaging strategies. For the intermediate-risk children with migraine headache and normal neurologic examination (prior probability of brain tumor 0.4%), CT-MRI was the most effective strategy but cost >$1 million per QALY gained compared with no neuroimaging. For high-risk children with headache of <6 months' duration and other clinical predictors of a brain tumor such as an abnormal neurologic examination (prior probability of brain tumor 4%), the most effective strategy was MRI, with cost-effectiveness ratio of $113 800 per QALY gained compared with no imaging. CONCLUSION: Our analysis suggests that MRI maximizes QALY gained at a reasonable cost-effectiveness ratio in children with headache at high risk of having a brain tumor. Conversely, the strategy of no imaging with close clinical follow-up is cost saving in low-risk children. Although the CT-MRI strategy maximizes QALY gained in the intermediate risk patients, its additional cost per QALY gained is high. In children with headache, appropriate selection of patients and diagnostic strategy may maximize quality-adjusted life expectancy and decrease costs of medical workup. PMID- 11483786 TI - Parents' Perceptions of Primary Care: measuring parents' experiences of pediatric primary care quality. AB - OBJECTIVE: A measure of pediatric primary care quality that is brief, practical, reliable, and valid would be useful to patients and pediatricians, policymakers, and health system leaders. Parents have a unique perspective from which to report their experiences with their child's primary care, and these reports may be valid indicators of pediatric primary care quality. The research objective was to develop a brief parent report of their children's primary care, the Parent's Perceptions of Primary Care measure (P3C), and to test its reliability and validity as a measure of pediatric primary care quality. STUDY DESIGN: The P3C was based on the elements of primary care as defined by the Institute of Medicine. Pretesting of domain content and item clarity was accomplished via focus interviews. The P3C was developed in English and translated to Spanish, Vietnamese, and Tagalog. The 23-item P3C yields a total score, as well as subscale scores for continuity, access, contextual knowledge, communication, comprehensiveness, and coordination. The P3C was administered to 3371 parents of children in kindergarten through sixth grades in a large, urban school district. PRINCIPAL FINDINGS: The percentage of missing values for the overall sample was 1.88%, indicating acceptable feasibility. Range of measurement, assessed via floor and ceiling effects, was moderate to good. Cronbach's coefficient alpha, an indicator of scale internal consistency reliability, was 0.95 for the P3C total scale. Factor analysis supported the subscale structure, and P3C scores were higher for children with health insurance, whose parents completed the survey in English, and who had a regular physician. P3C scores were positively related to parent reports of the child's health-related quality of life. CONCLUSIONS: The P3C is a practical, reliable, and valid measure of parents' reports of pediatric primary care quality. This brief measure could be used alone, or in conjunction with other measures, to enhance outcomes and evaluate the impact of systems changes on the delivery of the main elements of primary care. PMID- 11483787 TI - Recent skin injuries in normal children. AB - OBJECTIVE: The objective of our study was to collect data on the totality of recent skin injuries in normal children and adolescents, and to determine the relationship between the number of injuries, the age of the child, and the time of year in a temperate climate. METHODS: The participants in this study were children and adolescents seen successively for a reason other than trauma over a period of 1 year, by the first author (J.L.), in a university medical center in Quebec City, Canada. The total body surface, with the exception of the anal genital area, was examined systematically. The characteristics and location of all recent injuries (bruises, abrasions, scratches, cuts, burns, etc) were recorded. Scars from old injuries were ignored. The statistical method used for comparison was the Fisher's exact test. RESULTS: Two thousand forty examinations were done on 1467 youngsters from 0 to 17 years of age. Nine hundred thirty-one examinations were done on boys and 1109 on girls. The majority of children 9 months and older (76.6%) had at least 1 recent skin injury, without a significant difference between the sexes. Seventeen percent of the total sample of children had at least 5 injuries, whereas 4% had 10 or more, <1% had 15 or more, and 0.2% had 20 or more. The sites involved were mostly the lower limbs. Less than 2% of the total sample of children had injuries to the thorax, abdomen, pelvis, or buttocks, and <1% to the chin, ears, or neck. The majority of injuries observed were bruises, regardless of the time of year. There were, however, more skin injuries during the summer and the proportion of abrasions was higher at this time of the year. The 0- to 8-month age group was unique from all points of view. Skin injuries were rare in this age group (11.4%); they did not vary with the season, and they were mainly on the head and the face. Their injuries were mostly scratches. Bruises were found in only 1.2% of this group. CONCLUSIONS: The majority of normal children (after the age of 9 months) and adolescents, who do not consult for trauma, had 1 or more recent skin injuries. These injuries, mostly bruises, are more prevalent in the summer in a region with a temperate climate and can be present on all parts of the body, although they are most frequently observed on the limbs, especially on the shins and knees. Even if there are no recognizable marks on the skin, physicians must pay particular attention to children who have injuries with other unusual characteristics (uncommon location, >/=15 injuries, bruises in a child <9 months of age, numerous injuries elsewhere than the lower limbs, numerous injuries in the cold seasons in a temperate climate, injuries other than bruises, abrasions or scratches) because they could be a sign of a bleeding disorder or physical abuse.bruising, child abuse, accidental injury. PMID- 11483788 TI - Asthma symptoms, morbidity, and antiinflammatory use in inner-city children. AB - BACKGROUND: Asthma is a major cause of morbidity that disproportionately affects inner-city children. For children with persistent asthma, defined as having asthma symptoms 3 or more days per week or 3 or more nights per month, national guidelines recommend the use of daily antiinflammatory agents. Despite these recommendations, antiinflammatory agents remain underused, particularly in inner city children with high asthma morbidity. OBJECTIVES: The objectives of our study were to determine: 1) whether persistent asthma symptoms in inner-city children are related to acute care utilization and to the frequency of acute exacerbations; 2) whether children with persistent asthma are receiving recommended daily antiinflammatory agents; and 3) whether antiinflammatory medication use relates to sociodemographic factors, caretaker self-efficacy, the frequency of primary care visits, and/or measures of quality asthma care. DESIGN AND METHODS: A 64-item telephone survey was administered between July 1996 and June 1997 to 219 parental caretakers of 2- to 12-year-old children who had been hospitalized with asthma at an inner-city medical center between January 1995 and February 1996. Persistent asthma symptoms were assessed by inquiring about the frequency of daily and nocturnal asthma symptoms over the last 4 weeks. Children's asthma severity was classified by applying the 1997 National Asthma Education and Prevention Program (NAEPP) Asthma Guidelines' severity classification scheme based on the frequency of asthma symptoms. Asthma morbidity was defined as the frequency of acute asthma exacerbations, emergency department visits and hospitalizations. Daily antiinflammatory medication use was compared by sociodemographic factors, caretaker self-efficacy, frequency of primary care visits, and measures of quality asthma home management. RESULTS: In this sample, quantifying persistent asthma symptoms and applying the NAEPP symptom criteria identified 17% of the children with mild intermittent asthma, 27% with mild persistent asthma and 56% with moderate to severe persistent asthma. There were no differences in the age of the children in the 3 groups (mean age: 6 years). Asthma morbidity, as measured by the number of asthma exacerbations in the last 6 months, was significantly higher in the children with moderate to severe persistent asthma compared with children with mild intermittent asthma (9.8 vs 3.5) or mild persistent asthma (9.8 vs 4.5). In addition, there were significantly more emergency department visits in the moderate to severe group than in the mild persistent (3.05 vs 1.69) or mild intermittent group (3.05 vs 1.76). Lastly, as asthma symptom frequency increased, there were trends toward more hospitalizations and more days hospitalized. Overall, 35% of the 219 families reported giving daily antiinflammatory medications to their child (mostly cromolyn sodium). Of the 181 children (83%) who met NAEPP symptom criteria for persistent asthma, only 39% were receiving daily antiinflammatory treatment. Of the children with symptoms of moderate to severe asthma, only 15% were receiving inhaled steroids in contrast to the guidelines' recommendations. Use of antiinflammatory agents was not related to caretaker sociodemographic factors or self-efficacy scores. Measures of quality asthma home management, which included use of mattress covers, written plans, and peak flow meters, correlated positively with use of antiinflammatory agents. Children whose families reported using daily antiinflammatory medications had more primary care visits in the last 6 months than those children not receiving antiinflammatory medications. CONCLUSION: Questioning parents about the frequency of their child's asthma symptoms is an important, inexpensive, and readily accessible bedside and office tool that may aid in the detection of persistent symptoms and help direct therapy. Our study suggests that classifying asthma severity by quantifying persistent asthma symptoms, as defined in the NAEPP Guidelines, is a clinically useful tool that relates to asthma morbidity. In our sample of previously hospitalized children with asthma, 83% met 1997 NAEPP symptom criteria for persistent asthma, and yet only 35% were receiving daily antiinflammatory agents. Use of antiinflammatory agents correlated positively with other indicators of quality asthma home management. Additional work is necessary to increase appropriate use of antiinflammatory agents in this population, and in particular, to increase inhaled steroid use for children with moderate or severe symptoms. PMID- 11483789 TI - Switching to gatekeeping: changes in expenditures and utilization for children. AB - BACKGROUND: Gatekeeping has been a central strategy in the cost-containment initiatives of managed care organizations. Little empirical research describes the impact of switching into a gatekeeping plan on health care expenditures and utilization for children. OBJECTIVE: To determine the likelihood of a parent with a chronically ill child enrolling in a health plan with gatekeeping, as well as the effects of gatekeeping on health care expenditures and utilization for children, especially those with chronic conditions. DESIGN: We followed a cohort of 1839 children who either voluntarily switched to a gatekeeping plan or remained in an indemnity plan from 1991 through 1994. Study participants were children of employees of a large hospital. The gatekeeping plan was virtually identical to the previous indemnity plan except for lower monthly employee contribution and the requirement for a primary care physician to preapprove subspecialty referrals. We determined the likelihood of a household containing a child with a chronic condition enrolling in the gatekeeping plan, as well as mean annual total, subspecialty, and primary care expenditures and utilization for all children and children with chronic conditions. RESULTS: Households switching to gatekeeping were less likely to have children with chronic illness (8% vs 15%). Total and subspecialty expenditures for all children decreased more in the gatekeeping group (53% and 59%, respectively) than in the indemnity group (11% and 6%, respectively). For children with chronic conditions, mean visits to subspecialists decreased 57% in the gatekeeping group but increased 31% in the indemnity group. Mean visits to primary care physicians decreased 23% in the gatekeeping group compared with 13% in indemnity group. CONCLUSION: Parents of children with a chronic condition were much less likely than other parents to switch to a gatekeeping plan. Switching to gatekeeping was associated with reduced visits to specialists but did not increase the involvement of primary care physicians in the management of children with chronic conditions. The implications of these findings for the health of children are unknown. PMID- 11483790 TI - The decision to breastfeed in the United States: does race matter? AB - OBJECTIVES: To estimate the effects of maternal and birth characteristics on the decision to breastfeed and to relate breastfeeding practices to racial differences in infant mortality. METHODS: Using a sample of women with young children from the National Survey of Family Growth (NSFG), Cycle V, 1995, the likelihood of breastfeeding was modeled using logistic regression techniques. In addition, single, live births from the NSFG 1988 and 1995 surveys were analyzed to model the effects of race and breastfeeding on infant mortality using Cox regression methods. RESULTS: After controlling for socioeconomic background and birth characteristics, race remained a strong predictor of breastfeeding. Black women were less likely to breastfeed than nonblack women were, and the primary reason indicated by black women for not breastfeeding was that they "preferred to bottle-feed." Analyses of infant mortality indicated that breastfeeding accounts for the race difference in infant mortality in the United States at least as well as low birth weight does. CONCLUSIONS: Race is an important predictor of breastfeeding, with most black women reporting that they "preferred bottle feeding." Efforts to increase breastfeeding of infants in the black community should help narrow the racial gap in infant mortality. PMID- 11483791 TI - Effect of the Vaccines for Children program on physician referral of children to public vaccine clinics: a pre-post comparison. AB - OBJECTIVE: Started in late 1994, the Vaccines for Children (VFC) program is a major entitlement program that provides states with free vaccines for disadvantaged children. Some evaluation studies have been conducted, but they do not include individually matched pre-post comparisons of physician responses. This project studied the effect of the VFC on the physician likelihood of referring children to public vaccine clinics for immunizations. DESIGN: In 1999, trained personnel conducted a survey of a cohort of physicians who previously participated in surveys on barriers to childhood vaccination conducted before VFC implementation. Responses were matched, and pre- versus post-VFC comparisons were made. SETTING AND PARTICIPANTS: Minnesota and Pennsylvania primary care physicians selected by stratified random sampling and initially studied in 1990 to 1991 and 1993, respectively. MAIN OUTCOME MEASURES: Likelihood of referral of a child to a public vaccine clinic. RESULTS: On a scale of 0 to 10, physician likelihood of referring an uninsured child decreased by a mean of 1.9 (95% confidence interval: 1.2-2.5) from pre- to post-VFC. Two fifths (45%) of physicians reported that the VFC decreased the number of referrals from their practice to public vaccine clinics and 50% gave intermediate responses. Among physicians who participate in VFC, only 9% were likely to refer a Medicaid insured child in contrast to 44% of those not participating. CONCLUSIONS: Physicians' reported referral and likelihood of referring Medicaid-insured and uninsured children has decreased because of VFC in Minnesota and Pennsylvania.vaccination/economics, vaccination/legislation and jurisprudence, immunization programs/economics, immunization programs/utilization, vaccines/economics, Medicaid/economics, national health programs United States, child health services. PMID- 11483792 TI - Risk factors for positive mantoux tuberculin skin tests in children in San Diego, California: evidence for boosting and possible foodborne transmission. AB - OBJECTIVES: Source case finding in San Diego, California, rarely detects the source for children with tuberculosis (TB) infection or disease. One third of all pediatric TB isolates in San Diego are Mycobacterium bovis, a strain associated with raw dairy products. This study was conducted to determine risk factors for TB infection in San Diego. DESIGN: Case-control study of children /=10 mm) Mantoux skin test (TST) were matched by age to 1 to 2 children with negative TST from the same clinic. We assessed risk factors for TB infection through parental interview and chart review. RESULTS: A total of 62 cases and 97 controls were enrolled. Eleven cases and 25 controls were excluded from analysis because of previous positive skin tests. Compared with controls, cases were more likely to have received BCG vaccine (73% vs 7%, odds ratio [OR] 44), to be foreign born (35% vs 11%, OR 4.3), and to have eaten raw milk or cheese (21% vs 8%, OR 3.76). The median time between the most recent previous TST and the current test was 12 months for cases and 25 months for controls. Other factors associated with a positive TST included foreign travel, staying in a home while out of the country, and having a relative with a positive TST. There was no association between contact with a known TB case. In a multivariable model, receipt of BCG, contact with a relative with a positive TST, and having a previous TST within the past year were independently associated with TB infection. CONCLUSIONS: We identified several new or reemerging associations with positive TST including cross border travel, staying in a foreign home, and eating raw dairy products. The strong associations with BCG receipt and more recent previous TST may represent falsely positive reactions, booster phenomena, or may be markers for a population that is truly at greater risk for TB infection. Unlike studies conducted in nonborder areas, we found no association between positive TB skin tests and contact with a TB case or a foreign visitor. Efforts to control pediatric TB in San Diego need to address local risk factors including consumption of unpasteurized dairy products and cross-border travel. The interpretation of a positive TST in a young child in San Diego who has received BCG is problematic. PMID- 11483793 TI - Predictive model for serious bacterial infections among infants younger than 3 months of age. AB - OBJECTIVE: To develop a data-derived model for predicting serious bacterial infection (SBI) among febrile infants <3 months old. METHODS: All infants /=38.0 degrees C seen in an urban emergency department (ED) were retrospectively identified. SBI was defined as a positive culture of urine, blood, or cerebrospinal fluid. Tree-structured analysis via recursive partitioning was used to develop the model. SBI or No-SBI was the dichotomous outcome variable, and age, temperature, urinalysis (UA), white blood cell (WBC) count, absolute neutrophil count, and cerebrospinal fluid WBC were entered as potential predictors. The model was tested by V-fold cross-validation. RESULTS: Of 5279 febrile infants studied, SBI was diagnosed in 373 patients (7%): 316 urinary tract infections (UTIs), 17 meningitis, and 59 bacteremia (8 with meningitis, 11 with UTIs). The model sequentially used 4 clinical parameters to define high-risk patients: positive UA, WBC count >/=20 000/mm(3) or /=39.6 degrees C, and age <13 days. The sensitivity of the model for SBI is 82% (95% confidence interval [CI]: 78%-86%) and the negative predictive value is 98.3% (95% CI: 97.8%-98.7%). The negative predictive value for bacteremia or meningitis is 99.6% (95% CI: 99.4%-99.8%). The relative risk between high- and low-risk groups is 12.1 (95% CI: 9.3-15.6). Sixty-six SBI patients (18%) were misclassified into the lower risk group: 51 UTIs, 14 with bacteremia, and 1 with meningitis. CONCLUSIONS: Decision-tree analysis using common clinical variables can reasonably predict febrile infants at high-risk for SBI. Sequential use of UA, WBC count, temperature, and age can identify infants who are at high risk of SBI with a relative risk of 12.1 compared with lower-risk infants. PMID- 11483794 TI - Economic evaluation of a 2-dose hepatitis B vaccination regimen for adolescents. AB - OBJECTIVE: To investigate the economic implications of a 2-dose hepatitis B virus vaccination regimen compared with the current 3-dose vaccination regimen for adolescents in 3 settings: public schools, public health clinics, and private sector settings in the United States. METHODS: To measure resource utilization and costs associated with the administration of the 3-dose regimen and to assess vaccination compliance rates with this regimen, primary data were collected with the use of questionnaires tailored for each setting. Conservative modeling assumptions were used to derive 2-dose compliance rates from 3-dose compliance data. The results were incorporated into a decision analytic model, which was used to examine short-term and lifetime scenarios for an adolescent cohort receiving the 2-dose versus the 3-dose regimen. In the short-term analysis, the vaccination program costs were compared for the 2 regimens. In the lifetime analysis, the model also incorporated long-term disease costs for those individuals who contract hepatitis B. RESULTS: Predicted increases in compliance with a 2-dose vaccination regimen contributed to a higher probability of seroprotection in each setting. In the lifetime analysis, this positive impact of improved compliance resulted in a lower infection rate and greater cost effectiveness for the 2-dose regimen in all settings, including private sector settings, where it cost an average of only $964 per year of life gained, and in public schools, costing an average of $1246 per year of life gained. In public health clinics, the 2-dose regimen had both lower expected lifetime costs and better clinical outcomes than the 3-dose regimen. In the short-term analysis, costs were higher for the 2-dose regimen, reflecting higher total vaccine acquisition costs without the long-term offset of cost savings from reduced infection. Sensitivity analyses identified cost per dose of vaccine and the probability of completing the regimens as the most sensitive model variables. CONCLUSIONS: Improved compliance with a 2-dose regimen would contribute to a higher probability of adolescents' achieving seroprotection. When the long-term consequences of hepatitis B virus infection are included, the 2-dose regimen would be cost-effective compared with the 3-dose regimen in all settings and cost saving in public health clinic settings. PMID- 11483795 TI - Driveway injuries in children: risk factors, morbidity, and mortality. AB - BACKGROUND/PURPOSE: Injuries that occur around the driveway are not typically regarded as reportable to the police and thus are often underrecognized. The aim of this study was to characterize the pattern and consequences of motor vehicle collisions that occur in the driveway. METHODS: Over the past 13 years, 64 patients admitted to the Children's Hospital of Pittsburgh sustained motor vehicle-related injuries in a driveway. These injuries resulted from a vehicle driven by an adult driver striking a child (group 1) or a child shifting an idle vehicle out of gear (group 2). We compared demographic variables and outcome measures between the 2 groups. RESULTS: There was no difference in gender, injury pattern, Injury Severity Score, length of stay, or operations performed between the groups. Patients in group 1 were younger, smaller, had a lower Glasgow Coma Scale, and had poorer outcomes. The majority of collisions (~65%) in group 1 resulted from a truck or sport-utility vehicle going in reverse. CONCLUSIONS: Younger children are more severely injured in driveway-related crashes, which are most likely to be caused by a truck or sport-utility vehicle going in reverse. These vehicles should be equipped with additional safety features such as extended mirrors to visualize small children. PMID- 11483796 TI - Association between serum macrophage colony-stimulating factor levels and monocyte and thrombocyte counts in healthy, hypoxic, and septic term neonates. AB - OBJECTIVE: Macrophage colony-stimulating factor (M-CSF) is a hematopoietic growth factor that mainly stimulates the growth, differentiation, and proliferation of cells of the monocyte-macrophage lineage. There are only limited numbers of studies about M-CSF levels in neonates, but high levels of serum M-CSF have been reported in septic and some thrombocytopenic adult patients. In this study, we investigated the serum M-CSF levels in healthy, septic, and hypoxic term neonates on the first day of life and examined the relationship of serum M-CSF levels and circulating monocyte and thrombocyte counts in these newborn infants. STUDY DESIGN: Three groups were defined in this prospective study: group 1, healthy neonates with no risk factors (n = 40); group 2, neonates who had severe hypoxia (n = 20); and group 3, neonates who fulfilled the criteria for early-onset sepsis (n = 18). Blood samples were collected for complete blood cell count and serum M CSF levels by peripheral venipuncture from each infant in the first 24 hours after birth before any medical therapy. RESULTS: The gestational ages and birth weights did not differ significantly between the groups. Serum M-CSF levels of the septic neonates were significantly higher than of both healthy and hypoxic neonates, but did not differ significantly between the healthy and hypoxic neonates. There was no significant correlation between serum M-CSF levels and circulating monocyte counts, but there was a significant inverse correlation between serum M-CSF levels and thrombocyte counts. When this correlation was analyzed according to groups, we determined that this inverse correlation between M-CSF levels and thrombocyte counts was especially significant in the septic neonate group, but not significant in the healthy and hypoxic neonate groups. CONCLUSIONS: Serum M-CSF levels are significantly higher in neonates with sepsis. High serum M-CSF levels may have a possible role in the pathogenesis of thrombocytopenia in neonates with sepsis. PMID- 11483797 TI - Intention to return for papanicolaou smears in adolescent girls and young women. AB - OBJECTIVE: Sexually active adolescent girls have high rates of abnormal cervical cytology. However, little is known about factors that influence intention to return for Papanicolaou screening or follow-up. The aim of this study was to determine whether a theory-based model that assessed knowledge, attitudes, and behaviors predicted intention to return. METHODS: The study design consisted of a self-administered, cross-sectional survey that assessed knowledge, beliefs, perceived control over follow-up, perceived risk, cues for Papanicolaou smears, impulsivity, risk behaviors, and past compliance with Papanicolaou smear follow up. Participants were recruited from a hospital-based adolescent clinic that provides primary and subspecialty care, and the study sample consisted of all sexually active girls and young women who were aged 12 to 24 years and had had previous Papanicolaou smears. The main outcome measure was intention to return for Papanicolaou smear screening or follow-up. RESULTS: The enrollment rate was 92% (N = 490), mean age was 18.2 years, 50% were black, and 22% were Hispanic. Eighty-two percent of participants intended to return. Variables that were independently associated with intention to return included positive beliefs about follow-up (odds ratio [OR]: 1.07; 95% confidence interval [CI]: 1.02-1.11), perception that important others believe that the participant should obtain a Papanicolaou smear (OR: 1.93; 95% CI: 1.38-2.74), perceived control over returning (OR: 1.24; 95% CI: 1.06-1.46), and having received cues to obtain a Papanicolaou smear (OR: 1.31; 95% CI: 1.08-1.60). CONCLUSIONS: Analysis of this novel theoretical framework demonstrated that knowledge and previous behaviors were not associated with intention to return for Papanicolaou smear screening and follow-up in this population of young women. However, modifiable attitudinal components, including personal beliefs, perception of others' beliefs, and cues to obtaining Papanicolaou smears, were associated with intention to return. PMID- 11483798 TI - Smoking during pregnancy and infantile colic. AB - OBJECTIVE: To evaluate the association between maternal smoking during pregnancy and infantile colic (IC). METHODS: A follow-up study of singleton infants delivered by Danish mothers at the Aarhus University Hospital from May 1991 to February 1992 and still living in the municipality of Aarhus at the age of 8 months was conducted. A total of 1820 mothers and their infants were included. Self-administered questionnaires were used to collect data on smoking, other lifestyle factors, and sociodemographic variables at 16 and 30 weeks of gestation and 8 months postpartum. IC was defined in the 8-month questionnaire and based on Wessel's criteria, except that we used only the crying criterion. RESULTS: IC was seen in 10.8% of all infants. We observed a twofold increased risk of IC among infants whose mothers smoked 15 or more cigarettes per day during their pregnancy(relative risk: 2.1; 95% confidence interval: 1.4-3.2) or in the postpartum period (relative risk: 2.0; 95% confidence interval: 1.3-3.1). Women who smoked continuously during pregnancy and the postpartum period had a relative risk of 1.5 (95% confidence interval: 1.1-2.0) of having a infant with IC compared with women who did not smoke during this period. Adjustment for maternal age, parity, marital status, alcohol intake, birth weight, gestational age, breastfeeding, caffeine intake postpartum, and paternal smoking did not change the effect measures. CONCLUSION: The results indicate that maternal smoking during pregnancy may increase the risk of IC. PMID- 11483799 TI - Earlier onset of puberty in girls: relation to increased body mass index and race. AB - OBJECTIVE: A recent study conducted by the Pediatric Research in Office Settings network provided evidence that girls in the United States, especially black girls, are starting puberty at a younger age than earlier studies had found, but the reasons for this are not known. Because nutritional status is known to affect timing of puberty and there is a clear trend for increasing obesity in US children during the past 25 years, it was hypothesized that the earlier onset of puberty could be attributable to the increasing prevalence of obesity in young girls. Therefore, the objective of this study was to reexamine the Pediatric Research in Office Settings puberty data by comparing the age-normalized body mass index (BMI-ZS; a crude estimate of fatness) of girls who had breast or pubic hair development versus those who were still prepubertal, looking at the effects of age and race. RESULTS: For white girls, the BMI-ZS were markedly higher in pubertal versus prepubertal 6- to 9-year-olds; for black girls, a smaller difference was seen, which was significant only for 9-year-olds. Higher BMI-ZS also were found in girls who had pubic hair but no breast development versus girls who had neither pubic hair nor breast development. A multivariate analysis confirms that obesity (as measured by BMI) is significantly associated with early puberty in white girls and is associated with early puberty in black girls as well, but to a lesser extent. CONCLUSIONS: The results are consistent with obesity's being an important contributing factor to the earlier onset of puberty in girls. Factors other than obesity, however, perhaps genetic and/or environmental ones, are needed to explain the higher prevalence of early puberty in black versus white girls. PMID- 11483800 TI - Comparative practice patterns of emergency medicine physicians and pediatric emergency medicine physicians managing fever in young children. AB - BACKGROUND/OBJECTIVE: The management of fever in young children is a controversial topic. This study seeks to compare the management approaches between general emergency medicine physicians (GEMPs) and pediatric emergency medicine physicians (PEMPs) and correlate them to existing practice guidelines. DESIGN/METHODS: All charts of children age 3 to 36 months presenting with the complaint of fever at both a children's hospital emergency department (ED) and a general ED from June 1, 1998 to September 1, 1998; December 1, 1998 to April 1, 1999; and June 1, 1999 to September 1, 1999 were retrospectively reviewed. Fever was defined as >/=39 degrees C. Patients with a history of immunodeficiency, chronic illness, ventriculoperitoneal shunt, antibiotic use in the past 48 hours, or focal infection noted on examination were excluded. Data collected included focal exam findings, laboratory tests, diagnosis, treatment, and disposition. Variances from the practice guidelines were tabulated and compared. RESULTS: One thousand three hundred twenty-three eligible children met exclusion criteria and were seen by PEMPs; 755 were eliminated because of exclusion criteria (526 because of focal infection). Twenty-two (4%) of 568 remaining patients were admitted to the hospital. Two hundred twenty-eight eligible children were seen by GEMPs; 147 were excluded (109 because of focal infection). No patients were admitted to the hospital. PEMPs ordered more complete blood counts (324/568 vs 27/81), more blood cultures (321/568 vs 27/81), and more urine cultures (208/568 vs 20/81) than GEMPs. GEMPs ordered more chest radiographs and cerebrospinal fluid analyses than PEMPs; GEMPs ordered less complete blood counts, blood cultures, and urine cultures than PEMPs. GEMPs diagnosed more focal infections (109/228 vs 526/1323), and conflicted more often with the practice guidelines (66/79 vs 225/498) than PEMPs. Patients spent an average of 2.26 +/- 0.16 hours in the pediatric ED versus 3.0 hours +/- 0.18 hours in the general ED. CONCLUSIONS: Significant differences in the management of the young child with fever and no source exist between these two groups of physicians. These variations affect both cost and standard of care. Future studies assessing whether these strategies affect patient outcomes would further elucidate their clinical implication. PMID- 11483801 TI - Growth and development in preterm infants fed long-chain polyunsaturated fatty acids: a prospective, randomized controlled trial. AB - OBJECTIVES: A randomized, masked, controlled trial was conducted to assess effects of supplementing premature infant formulas with oils containing the long chain polyunsaturated fatty acids, arachidonic acid (AA; 20:4 n6), and docosahexaenoic acid (DHA; 22:6 n3) on growth, visual acuity, and multiple indices of development. METHODS: Infants (N = 470) with birth weights 750 to 1800 g were assigned within 72 hours of the first enteral feeding to 1 of 3 formula groups with or without long-chain polyunsaturated fatty acids: 1) control (N = 144), 2) AA+DHA from fish/fungal oil (N = 140), and 3) AA+DHA from egg-derived triglyceride (egg-TG)/fish oil (N = 143). Infants were fed human milk and/or Similac Special Care with or without 0.42% AA and 0.26% DHA to term corrected age (CA), then fed human milk or NeoSure with or without 0.42% AA and 0.16% DHA to 12 months' CA. Infants fed exclusively human milk to term CA (EHM-T; N = 43) served as a reference. RESULTS: Visual acuity measured by acuity cards at 2, 4, and 6 months' CA was not different among groups. Visual acuity measured by swept parameter visual-evoked potentials in a subgroup from 3 sites (45 control, 50 AA+DHA [fish/fungal]; 39 AA+DHA [egg-TG/fish]; and 23 EHM-T) was better in both the AA+DHA (fish/fungal; least square [LS] means [cycle/degree] +/- standard error [SE; octaves] 11.4 +/- 0.1) and AA+DHA (egg-TG/fish; 12.5 +/- 0.1) than control (8.4 +/- 0.1) and closer to that of the EHM-T group (16.0 +/- 0.2) at 6 months' CA. Visual acuity improved from 4 to 6 months' CA in all but the control group. Scores on the Fagan test of novelty preference were greater in AA+DHA (egg TG/fish; LS means +/- SE, 59.4 +/- 7.7) than AA+DHA (fish/fungal; 57.0 +/- 7.5) and control (57.5 +/- 7.4) at 6 months' CA, but not at 9 months' CA. There were no differences in the Bayley Mental Development Index at 12 months' CA. However, the Bayley motor development index was higher for AA+DHA (fish/fungal; LS means +/- SE, 90.6 +/- 4.4) than control (81.8 +/- 4.3) for infants /=10 mm, or history of tuberculosis family contact. Patients with definitive or probable PTB were significantly older than patients without clinical PTB, and those with symptomatic disease were significantly older than those with asymptomatic disease. Patients with PTB diagnosed by culture were significantly more likely than those diagnosed using clinical criteria to have cough lasting >/=2 weeks, fever, and a PPD >/=10 mm. CONCLUSIONS: The typical presentation of PTB in Peruvian children includes symptoms of active pulmonary disease similar to those seen in adults. This presentation differs significantly from that reported in developed countries, where many children have minimal or no symptoms at the time of presentation. The diagnostic criteria for pediatric PTB must be modified in hyperendemic developing country environments where features may differ from those described in the United States. The triad of cough lasting >/=2 weeks, fever, and a PPD >/=10 mm was highly predictive for culture-positive PTB among children in this low-income Peruvian population. PMID- 11483815 TI - More research is needed to determine the effects of human milk on neonatal outcome. PMID- 11483816 TI - Balancing risks and benefits: Primum non nocere is too simplistic. PMID- 11483817 TI - Rescue use of DNase in critical lung atelectasis and mucus retention in premature neonates. AB - Excessive or thick pulmonary secretions are a common clinical challenge in the neonatal population. Mucus accumulation can cause many life-threatening complications, including plugging of the endotracheal tube and increasing the risk of pulmonary infections. We report 3 premature neonates who had critical pulmonary collapse secondary to mucous plugging. Different conventional methods to liquefy mucus and facilitate removal of secretions were exhausted to no avail. The rescue use of DNase was effective in reestablishing airway patency. Thus, this drug could be a valuable tool in treating atelectasis and mucus-plugging in mechanically ventilated, premature neonates. PMID- 11483818 TI - A 1000-fold overdose of clonidine caused by a compounding error in a 5-year-old child with attention-deficit/hyperactivity disorder. AB - A 5-year-old child who weighed 17.5 kg received 50 mg of clonidine. The amount ingested was confirmed by analysis of the suspension administered (clonidine HCl 9.78 mg/mL). To our knowledge, this represents the largest ingestion in a child and the largest ingestion on a milligram per kilogram basis in the medical literature. The child's initial presentation included hyperventilation, an unusual feature of clonidine toxicity. The child was discharged without sequela 42 hours after admission. A serum concentration of clonidine 17 hours postingestion was 64 ng/mL, the highest reported to date in a pediatric patient. The intoxication was traced to a pharmacy compounding error in which milligrams were substituted for micrograms. Increased prescribing of clonidine in young children coupled with the requirement to compound clonidine in a suspension and the narrow therapeutic index suggests that the frequency of severe ingestions in children will increase in the future. PMID- 11483819 TI - Methyl ethyl ketone peroxide ingestion: toxicity and outcome in a 6-year-old child. AB - A 6-year-old boy developed respiratory distress, metabolic acidosis, severe esophageal and gastric burns, and a coagulopathy after ingestion of an unknown volume of methyl ethyl ketone peroxide (MEKP) in dimethyl phthalate. He was discharged from the pediatric intensive care unit 19 days postingestion but subsequently developed a stricture of the gastroesophageal junction and complete fibrosis of the middle third of the stomach, necessitating gastric resection and reconstruction. He was discharged 93 days postingestion on a program of dilation for the residual esophageal stricture. MEKP acts by initiating lipid peroxidation via free radical production that results in cellular dysfunction and death. Acetylcysteine, a glutathione precursor and possible free radical scavenger, may be of use in severe MEKP poisoning. This case demonstrates the severe effects that some industrial chemicals can have both systemically and locally at the point of contact with the gastrointestinal tract, as well as the long-term management required to ensure good quality of life. PMID- 11483820 TI - Optic neuropathy in children with Lyme disease. AB - Involvement of the optic nerve, either because of inflammation or increased intracranial pressure, is a rare manifestation of Lyme disease. Of the 4 children reported here with optic nerve abnormalities, 2 had decreased vision months after disease onset attributable to optic neuritis, and 1 had headache and diplopia early in the infection because of increased intracranial pressure associated with Lyme meningitis. In these 3 children, optic nerve involvement responded well to intravenous ceftriaxone therapy. The fourth child had headache and visual loss attributable to increased intracranial pressure and perhaps also to optic neuritis. Despite treatment with ceftriaxone and steroids, he had persistent increased intracranial pressure leading to permanent bilateral blindness. Clinicians should be aware that neuro-ophthalmologic involvement of Lyme disease may have significant consequences. If increased intracranial pressure persists despite antibiotic therapy, measures must be taken quickly to reduce the pressure. PMID- 11483821 TI - Treatment of pain with gabapentin in a neonate. PMID- 11483822 TI - Drug-induced hypersensitivity syndrome in pediatric patients. AB - The antiepileptic hypersensitivity syndrome is a severe, multiorgan reaction to oral antiepileptics that manifests as fever, rash, lymphadenopathy, and hepatitis. This same reaction pattern also has been described following administration of a few unrelated medications. We report on 11 patients who had drug-induced hypersensitivity syndrome and were admitted to our pediatric service and review 94 cases of this syndrome in pediatric patients identified from the literature. We undertook this study to summarize the findings and alert clinicians to the severe internal organ involvement that can occur with this syndrome. PMID- 11483823 TI - Delayed separation of the umbilical cord attributable to urachal anomalies. PMID- 11483824 TI - Case report: liver glycogen synthase deficiency--a cause of ketotic hypoglycemia. AB - Glycogen synthase deficiency is a rare inborn error of metabolism, characterized by fasting hypoglycemia, hypoglycemic seizures, and ketonuria. Only 7 families with 14 affected children have been reported. Here, we report an additional patient with this deficiency. Findings in this patient were clinically and biochemically consistent with those reported in patients with ketotic hypoglycemia and may alert the clinician to consider glycogen synthase deficiency. PMID- 11483825 TI - American Academy of Pediatrics: Committee on Psychosocial Aspects of Child and Family Health and Committee on Adolescence. Sexuality education for children and adolescents. AB - Children and adolescents need accurate and comprehensive education about sexuality to practice healthy sexual behavior as adults. Early, exploitative, or risky sexual activity may lead to health and social problems, such as unintended pregnancy and sexually transmitted diseases, including human immunodeficiency virus infection and acquired immunodeficiency syndrome. This statement reviews the role of the pediatrician in providing sexuality education to children, adolescents, and their families. Pediatricians should integrate sexuality education into the confidential and longitudinal relationship they develop with children, adolescents, and families to complement the education children obtain at school and at home. Pediatricians must be aware of their own attitudes, beliefs, and values so their effectiveness in discussing sexuality in the clinical setting is not limited. PMID- 11483826 TI - American Academy of Pediatrics: Technical report: knee brace use in the young athlete. AB - This statement is a revision of a previous statement on prophylactic knee bracing and provides information for pediatricians regarding the use of various types of knee braces, indications for the use of knee braces, and the background knowledge necessary to prescribe the use of knee braces for children. PMID- 11483827 TI - American Academy of Pediatrics: Committee on Child Abuse and Neglect and Committee on Children With Disabilities. Assessment of maltreatment of children with disabilities. AB - Widespread efforts are continuously being made to increase awareness and provide education to pediatricians regarding risk factors of child abuse and neglect. The purpose of this statement is to ensure that children with disabilities are recognized as a population that is also at risk for maltreatment. The need for early recognition and intervention of child abuse and neglect in this population, as well as the ways that a medical home can facilitate the prevention and early detection of child maltreatment, should be acknowledged. PMID- 11483828 TI - American Academy of Pediatrics: Task Force on Medical Informatics. Special requirements for electronic medical record systems in pediatrics. AB - Electronic medical record (EMR) systems, which are usually designed for adult care, must perform certain functions to be useful in pediatric care. This statement outlines these functions (eg, immunization tracking and pediatric dosing calculations) to assist vendors and standards organizations with software design for pediatric systems. The description of these functions should also provide pediatricians with a set of requirements or desirable features to use when evaluating EMR systems. Particular attention is paid to special aspects of pediatric clinical care and privacy issues unique to pediatrics. PMID- 11483829 TI - American Academy of Pediatrics: Committee on Injury and Poison Prevention. School bus transportation of children with special health care needs. AB - School systems are responsible for ensuring that children with special needs are safely transported on all forms of federally approved transportation provided by the school system, and a plan should be developed to provide the most current and proper support to children with special transportation requirements. This statement provides current guidelines for the protection of child passengers with specific health care needs, including those with a tracheostomy, those requiring use of car seats, or those transported in wheelchairs. Guidelines that apply to general school transportation should be followed, including the training of staff, provision of nurses or aides if needed, establishment of a written emergency evacuation plan, and a comprehensive infection control program. Research provides the basis for recommendations concerning occupant securement for children in wheelchairs and children with other special needs who are transported on a school bus. Pediatricians can help their patients by being aware of guidelines for restraint systems for children with special needs and by remaining informed of new resources. Pediatricians can also play an important role at the state and local level in the development of school bus specifications. PMID- 11483830 TI - A prospective multicenter study of cervical spine injury in children. AB - OBJECTIVE: Pediatric victims of blunt trauma have developmental and anatomic characteristics that can make it difficult to assess their risk of cervical spine injury (CSI). Previous reports, all retrospective in nature, have not identified any cases of CSI in either children or adults in the absence of neck pain, neurologic symptoms, distracting injury, or altered mental status. The objective of this study was to examine the incidence and spectrum of spine injury in patients who are younger than 18 years and to evaluate the efficacy of the National Emergency X-Radiography Utilization Study (NEXUS) decision instrument for obtaining cervical spine radiography in pediatric trauma victims. METHODS: We performed a prospective, multicenter study to evaluate pediatric blunt trauma victims. All patients who presented to participating emergency departments underwent clinical evaluation before radiographic imaging. The presence or absence of the following criteria was noted: midline cervical tenderness, altered level of alertness, evidence of intoxication, neurologic abnormality, and presence of painful distracting injury. Presence or absence of each individual criterion was documented for each patient before radiographic imaging, unless the patient was judged to be too unstable to complete the clinical evaluation before radiographs. The decision to radiograph a patient was entirely at the physician's discretion and not driven by the NEXUS questionnaire. The presence or absence of CSI was based on the final interpretation of all radiographic studies. Data on all patients who were younger than 18 years were sequestered from the main database for separate analysis. RESULTS: There were 3065 patients (9.0% of all NEXUS patients) who were younger than 18 years in this cohort, 30 of whom (0.98%) sustained a CSI. Included in the study were 88 children who were younger than 2, 817 who were between 2 and 8, and 2160 who were 8 to 17. Fractures of the lower cervical vertebrae (C5-C7) accounted for 45.9% of pediatric CSIs. No case of spinal cord injury without radiographic abnormality was reported in any child in this study, although 22 cases were reported in adults. Only 4 of the 30 injured children were younger than 9 years, and none was younger than 2 years. Tenderness and distracting injury were the 2 most common abnormalities noted in patients with and without CSI. The decision rule correctly identified all pediatric CSI victims (sensitivity: 100.0%; 95% confidence interval: 87.8%-100.0%) and correctly designated 603 patients as low risk for CSI (negative predictive value: 100.0%; 95% confidence interval: 99.4%-100.0%). CONCLUSIONS: The lower cervical spine is the most common site of CSI in children, and fractures are the most common type of injury. CSI is rare among patients aged 8 years or younger. The NEXUS decision instrument performed well in children, and its use could reduce pediatric cervical spine imaging by nearly 20%. However, the small number of infants and toddlers in the study suggests caution in applying the NEXUS criteria to this particular age group. PMID- 11483831 TI - Brain white-matter lesions in celiac disease: a prospective study of 75 diet treated patients. AB - OBJECTIVE: Celiac disease (CD), or gluten sensitivity, is considered to be a state of heightened immunologic responsiveness to ingested gluten proteins in genetically predisposed individuals. The gastrointestinal manifestation suggests a severe enteropathy of the small intestine with malabsorption, steatorrhea, and weight loss because of a deranged mucosal immune response. Neurologic complications occur, especially epilepsy, possibly associated with occipital calcifications or folate deficiency and cerebellar ataxia. There have been reports of brain white-matter lesions as an extraintestinal manifestation in Crohn disease and ulcerative colitis but not in CD. METHODS: In this study, 75 diet-treated mainly pediatric patients with biopsy-proven CD underwent prospectively clinical neurologic examinations, laboratory investigations, electroencephalography, computed tomography, and magnetic resonance imaging. The age range was 2.8 to 24.2 years with a mean of 11.6 years. The mean period of gluten exposure was 2.4 years. RESULTS: Ten patients had neurologic findings such as febrile seizures, single generalized seizures, mild ataxia, and muscular hypotonia with retarded motor development. No folate deficiency was found. The hippocampal regions showed no abnormalities. Computed tomography did not reveal any cerebral calcifications, but magnetic resonance imaging detected unilateral and bilateral T2-hyperintensive white-matter lesions in 15 patients (20%). There was no correlation between these lesions and dietary compliance or neurologic or electroencephalographic abnormalities. The mean gluten exposure time of these patients was slightly increased (not significant). CONCLUSIONS: Focal white matter lesions in the brain may represent an extraintestinal manifestation of CD. They may be ischemic in origin as a result of a vasculitis or caused by inflammatory demyelination. They seem to be more typical of pediatric CD than cerebral calcifications. Their prognostic value is unclear and needs to be elucidated in additional studies. CD should be suggested as a differential diagnosis in children with unclear white-matter lesions even without intestinal symptoms. PMID- 11483832 TI - Survey of pediatrician practices in retrieving statewide authorized newborn screening results. AB - OBJECTIVE: Mandated state newborn screening programs for the approximately 4 million infants born each year in the United States involves the following 5 components: 1) initial screening, 2) immediate follow-up testing of the screen positive newborn, 3) diagnosis confirmation (true positive versus false positive), 4) immediate and long-term care, and 5) evaluation of all of the components of the system, including process and outcomes measures. Smooth functioning of this system requires pretest education of the parents as well as education and involvement of all health care providers who interact with the newborn screening system. Although extensive literature is available concerning public health aspects, technical standards/protocols, and discussion of the interfaces among the 5 components of the system, little information is available regarding physician awareness, involvement, and interactions with the system. The objective of this study was to determine, through a survey, primary care pediatricians' satisfaction with their state's newborn screening program. This was reflected in survey questions that asked how pediatricians were notified of the results of newborn screening tests that were performed on infants in their practice. METHODS: Two thousand questionnaires were sent to primary care pediatricians in all 50 states and the District of Columbia regarding their practices in retrieving statewide newborn screening results. Of the 2000 surveys, 574 (29%) responses from primary care pediatricians who care for at least 1 to 5 newborns each week form the basis of this report. Also reported are the commentaries of the physicians concerning their specific practices, overall assessment of the system, and ideas for improvement. RESULTS: Physicians reported their general satisfaction with the newborn screening system's ability to retrieve screen-positive infants for follow-up testing. However, communication and partnership with the primary care pediatrician regarding accessibility and timely retrieval of newborn screening test results was deemed less than optimal. Thirty-one percent of respondents indicated that notification for screen-positive test results was greater than 10 days, whereas 26% indicated that they do not receive the results of screen-negative tests and need to develop office procedures (contact birth hospital or state laboratory) to obtain results. Twenty eight percent indicated that they do not actively seek results of newborn screening for their patients and presume that "no news is good news." Barriers to retrieving test results included that infants were born at hospitals where the physician does not have privileges, there were new transfers to the practice, infants were born in other states, personnel time was needed to track results, and there was a lack of a cohesive communication/reporting system that includes the primary care physician as an integral partner in the newborn screening communication process. Ninety-two percent of physicians would welcome an enhanced state system with direct communication to the primary care pediatrician as well as the birth hospital. CONCLUSION: Pediatricians recognize and endorse the benefits of newborn screening and believe that they play an important role in the efficient functioning of the system. An enhanced physician partnership with the newborn screening program will enable the timely follow-up of the screen-positive newborn for confirmatory testing. All test results need to be communicated to the pediatrician in a timely and efficient manner: 7 days for screen-positive results and 10 to 14 days for all results. Newborn screening test results of new patients who enter the practice should be available at the time of the first well-infant visit, ideally by 2 weeks of age. The majority of primary care pediatricians acknowledge the need to establish office protocols for the retrieval of newborn screening test results and would welcome an enhanced direct communication system with the state newborn screening program. PMID- 11483833 TI - Serotype prevalence of occult pneumococcal bacteremia. AB - OBJECTIVE: The licensure and use of a pneumococcal conjugate vaccine that is immunogenic in children who are younger than 2 years may affect the epidemiology of occult bacteremia. This study was conducted to determine the serotype prevalence of Streptococcus pneumoniae isolates from children with occult bacteremia and to document the proportion that would be covered by the recently licensed heptavalent pneumococcal conjugate vaccine. METHODS: A cohort of 5901 children who were 2 to 24 months of age and had a temperature of >/=39.0 degrees C evaluated with a blood culture at an urban tertiary care children's hospital emergency department was studied to determine the prevalence of S pneumoniae serotypes. Patients were excluded if their immune system was suppressed, they had a diagnosis of a focal infection, they were evaluated by lumbar puncture, they were admitted to the hospital, or they died during initial evaluation. Blood cultures were inoculated into pediatric blood culture bottles and processed using an automated carbon dioxide monitoring system. All pneumococcal isolates were serotyped on the basis of capsular swelling with type-specific antisera (Quellung reaction). RESULTS: The study population consisted of 5901 patients. The overall rate of occult bacteremia was 1.9% (95% confidence interval [CI]: 1.5-2.3). S pneumoniae accounted for 92 of 111 isolates (82.9%; 95% CI: 74.6-89.4) in children with occult bacteremia. Eight pneumococcal serotypes were represented: 6A (2%), 9V (6%), 19F (6%), 18C (8%), 4 (9%), 6B (13%), 23F (15%), and 14 (42%). Serotypes 14, 6B, and 23F accounted for 69.3% (95% CI: 58.6-78.7) of typed isolates. In the cohort, 97.7% (95% CI: 92-99.7) of isolated serotypes are represented in the newly licensed heptavalent pneumococcal conjugate vaccine. The single isolated serotype that would not have been covered by the currently licensed heptavalent pneumococcal conjugate vaccine was 6A. CONCLUSIONS: S pneumoniae accounts for the vast majority of bacterial pathogens in children with occult bacteremia. As indicated by the results of this study, the heptavalent pneumococcal conjugate vaccine may prevent the majority of occult pneumococcal bacteremia episodes. The 2 cases of bacteremia with a serotype that would not have been included in the vaccine both were due to serotype 6A. It has been noted that there is potential nonvaccine serotype and subgroup cross-protection (6A from 6B) afforded to children who are immunized with the heptavalent vaccine. The high potential efficacy of the heptavalent pneumococcal conjugate vaccine for strains that cause occult bacteremia in our population may have a profound effect on the treatment of children with fever without a source. There has been an alarming and rapid emergence of antibiotic-resistant pneumococcal strains. Less pressure to use broad-spectrum antibiotics, which in turn causes further antibiotic resistance, should result. Laboratory testing and hospitalization also should be reduced. The prevalence rates determined by this study may be used as baseline data for comparison of serotype rates of occult pneumococcal bacteremia after widespread use of the heptavalent vaccine. PMID- 11483835 TI - Psychotropic medication in children: a study from the Netherlands. AB - OBJECTIVE: Although there is a global concern about the increased use of psychotropic agents in children, most research literature originates in the United States and is based on figures from the first half of the 1990s. Also, few studies document the use of various types of psychotropic agents. The objective of this study was to investigate the use of psychotropic medication in children in the Netherlands and to determine whether this corresponds with previously reported figures from the United States. METHODS: A drug utilization study based on computerized pharmacy dispensing records was conducted from 1995 to 1999 for children aged 0 to 19 years in the north of the Netherlands. RESULTS: Stimulants were the most widely used psychotropic agents among 0- to 19-year-olds (prevalence: 7.4/1000 in 1999), followed by hypnotics/anxiolytics (6.9/1000) and antidepressants (4.4/1000). Prevalence rates of stimulants increased from 1.5/1000 in 1995 to 7.4/1000 in 1999. Incidence rates, proportion of girls, and duration of stimulant treatment increased as well. Changes in prevalence rates of other psychotropic agents were much smaller than those of stimulants. Finally, the vast majority of children who were treated with psychotropic agents used only 1 agent at a time. CONCLUSION: The prevalence of stimulant use in the Netherlands is much lower than reported previously (28/1000 children in 1995) from the United States, and differences also existed with regard to the use of other psychotropic agents and combinations of psychotropic agents. However, the increase in Dutch stimulant use agrees with the previously reported 2.5-fold increase in the United States and shows that the increased use of stimulants is not limited to the United States. PMID- 11483834 TI - Cost-effectiveness analysis of an intranasal influenza vaccine for the prevention of influenza in healthy children. AB - OBJECTIVE: Intranasal influenza vaccine has proven clinical efficacy and may be better tolerated by young children and their families than an injectable vaccine. This study determined the potential cost-effectiveness (CE) of an intranasal influenza vaccine among healthy children. METHODS: We conducted a CE analysis of data collected between 1996 and 1998 during a prospective 2-year efficacy trial of intranasal influenza vaccine, supplemented with data from the literature. The CE analysis included both direct and indirect costs. We enrolled 1602 healthy children aged 15 to 71 months in year 1, 1358 of whom were enrolled in year 2. One or 2 doses of intranasal influenza vaccine or placebo were administered to measure the cost per febrile influenza-like illness (ILI) day avoided. RESULTS: During the 2-year study period, vaccinated children had an average of 1.2 fewer ILI fever days/child than unvaccinated children. In an individual-based vaccine delivery scenario with vaccine given twice in the first year and once each year thereafter at an assumed base case total cost of $20 for the vaccine and its administration (ie, per dose), CE was approximately $30/febrile ILI day avoided. CE ranged from $10 to $69/febrile ILI day avoided at $10 to $40/dose, respectively. In a group-based delivery scenario, vaccination was cost saving compared with placebo and remained so if vaccine cost was <$28 (the break-even price per dose). In the individual-based scenario, vaccination was cost saving if vaccine cost was <$5. In this scenario, nearly half of lost productivity in the vaccine group was attributable to vaccine visits, which overshadowed the relatively modest savings in ILI-associated costs averted. CONCLUSIONS: Routine use of intranasal influenza vaccine among healthy children may be cost-effective and may be maximized by using group-based vaccination approaches. cost effectiveness, influenza, vaccine, children. PMID- 11483836 TI - Chronic oxygen dependency in infants born at less than 32 weeks' gestation: incidence and risk factors. AB - OBJECTIVE: To assess incidence and clinical risk factors of chronic oxygen dependency (COD) among survivors who were born at or before 31 weeks' gestation. METHODS: This prospective, multicenter study enrolled 802 infants who were born at or before 31 weeks' gestation and admitted to 8 level III neonatal intensive care units in northern and eastern France from January 1 through December 31, 1997. Need for oxygen to maintain oxygen saturation between 92% and 96% was assessed at 28 days of life and at 36 and 42 weeks' postconceptional age (PCA). Stepwise logistic regression analysis was used to identify the incidence of COD and the risk factors related to its occurrence. RESULTS: The mortality rate was 14%. Antenatal corticotherapy was administered to 51% of patients, surfactant therapy to 76% of the ventilated patients, and high-frequency oscillatory ventilation at day 1 to 32%. At 28 days and 36 and 42 weeks' PCA, respectively, 25%, 15%, and 6% of survivors had COD. After adjustment for intercenter variations, we identified the significant risk factors for COD at these dates: a low gestational age, a high score on the Clinical Risk Index for Infants, intrauterine growth restriction, and surfactant treatment. CONCLUSION: COD incidence was high at 28 days of life but decreased dramatically by 42 weeks' PCA. This study confirmed previously reported risk factors and underlined the importance of intrauterine growth restriction and the Clinical Risk Index for Infants as significant risk factors. PMID- 11483837 TI - Infection with Sin Nombre hantavirus: clinical presentation and outcome in children and adolescents. AB - OBJECTIVE: Sin Nombre hantavirus (SNV) is the leading causative agent of hantavirus cardiopulmonary syndrome (HCPS) in the United States and Canada. Relatively few cases of HCPS have involved children. This report describes the clinical characteristics of a series of pediatric cases of SNV infection in the United States and Canada from 1993 through March 2000. METHODS: We analyzed clinical and laboratory data on 13 patients who were 85% of patients had elevated levels of serum aspartate aminotransferase, alanine aminotransferase, and hypoalbuminemia. Leukocytosis and hemoconcentration were seen in less than one third of patients at admission. HCPS developed in 12 of the 13 patients (92%), and 4 of those 12 died (33% case-fatality ratio). The majority of HCPS patients (8 of 12 [67%]) were critically ill and required mechanical ventilation. Extracorporeal membrane oxygenation was used in 2 patients, 1 of whom survived. An elevated prothrombin time (>/=14 seconds) at admission was predictive of mortality. CONCLUSIONS: Infection with SNV in children and adolescents causes HCPS with a clinical course and mortality rate similar to that described in adults. We believe that early recognition of HCPS in children and adolescents and appropriate referral to tertiary care centers that are experienced with HCPS are important in reducing mortality. PMID- 11483838 TI - Predictors of mother-adolescent discussions about condoms: implications for providers who serve youth. AB - OBJECTIVE: To examine predictors of mother-adolescent communication about condoms. METHODS: Interviews were conducted with 907 mothers of adolescents aged 14 to 17 years in the Bronx, New York; Montgomery, Alabama; and San Juan, Puerto Rico, to determine whether mothers had talked with their adolescent about condoms. RESULTS: By univariate analysis, mother-adolescent communication about condoms was associated with greater knowledge about sexuality and acquired immunodeficiency syndrome, perception of having enough information to discuss condoms, information from a health-related source, less conservative attitudes about adolescent sexuality, perception that the adolescent was at risk for human immunodeficiency virus, greater ability and comfort in discussing condoms, stronger belief that condoms prevent human immunodeficiency virus/acquired immunodeficiency syndrome, and a more favorable endorsement of condoms. In multivariate analyses, mother-adolescent communication about condoms was associated with a less conservative attitude about abstinence until marriage (odds ratio [OR]: 0.73; 95% confidence interval [CI]: 0.54-0.74), greater skill in communicating about sex (OR: 1.13; 95% CI: 1.06-1.20), greater comfort in communicating about sex (OR: 1.31; 95% CI: 1.01-1.69), a more favorable endorsement of condoms (OR: 1.85; 95% CI: 1.17-2.78), and the perception that the adolescent's friends were sexually active (OR: 3.53; 95% CI: 1.97-7.16). CONCLUSION: Parents who communicate effectively about sexuality and safer sex behaviors can influence their adolescents' risk-taking behavior. Health care providers, particularly physicians, can facilitate this communication by providing to parents information about the sexual behavior of adolescents, the risks that adolescents encounter, condom use, condom effectiveness, and how to discuss condoms. They also can make referrals to programs that teach communication skills. PMID- 11483839 TI - A statewide assessment of lead screening histories of preschool children enrolled in a Medicaid managed care program. AB - OBJECTIVES: Despite the prominence of lead poisoning as a public health problem, recent Government Accounting Office reports indicate that only one fifth of children who are covered by Medicaid have been screened for lead poisoning. The purpose of this study was to examine the lead screening histories of children who were enrolled in a statewide, Medicaid managed care plan to determine the relative impact of the type of primary care provider site and family sociodemographic characteristics on the likelihood of being screened. The study also examined the prevalence of and risk factors for lead poisoning of children who had been screened. METHODS: A random sample of 2000 preschool-age children was chosen from those who were enrolled continuously in the statewide, expanded, Medicaid managed care program for a 1-year period and between the ages of 19 and 35 months at the end of that year. Sociodemographic characteristics and lists of primary care providers were obtained from administrative data sets. Medical record audits at primary care provider sites were performed to obtain the lead screening histories of the children, including test dates and results. RESULTS: Data on 1988 children were used for study analyses, and 80% of these children had at least 1 documented blood lead level. Children whose primary care provider was an office-based physician were less likely to be screened as compared with patients of health centers, hospital-based clinics, and staff model health maintenance organizations (68%, 86%, 89%, and 91% respectively). Variation in screening rates persisted in a multivariate analysis controlling for family sociodemographic characteristics and practice level variation. Of the 1587 children who had a documented blood lead test, 467 children (29%) had a blood lead level of >/=10 mg/dL on at least 1 test. CONCLUSIONS: Blood lead screening rates in Rhode Island's Medicaid managed care program are dramatically higher than national estimates for children who are enrolled in Medicaid. Potential explanations for this finding are 1) a high sensitization to the problem of lead poisoning in Rhode Island, 2) the primary care focus of the Medicaid program in Rhode Island facilitates the delivery of preventive services, and 3) the medical record audit approach used in this study was more comprehensive in identifying blood lead screens than techniques used in national studies. The high prevalence of elevated blood lead levels found in this study emphasizes the importance of screening among children who are enrolled in Medicaid. PMID- 11483840 TI - Adoption as a risk factor for attempted suicide during adolescence. AB - OBJECTIVE: Depression, impulsivity, and aggression during adolescence have been associated with both adoption and suicidal behavior. Studies of adopted adults suggest that impulsivity, even more than depression, may be an inherited factor that mediates suicidal behavior. However, the association between adoption and adolescent suicide attempts and the mechanisms that might explain it remain unknown. The objective of this study was to determine the following: 1) whether suicide attempts are more common among adolescents who live with adoptive parents rather than biological parents; 2) whether the association is mediated by impulsivity, and 3) whether family connectedness decreases the risk of suicide attempt regardless of adoptive or biological status. METHODS: A secondary analysis of Wave I data from the National Longitudinal Study of Adolescent Health was conducted, which used a school-based, clustered sampling design to identify a nationally representative sample of 7th- to 12th-grade students, with oversampling of underrepresented groups. Of the 90 118 adolescents who completed the National Longitudinal Study of Adolescent Health in-school survey, 17 125 completed the in-home interview and had parents of identified gender who completed separate in-home questionnaire. The subset of adolescents for this study was drawn from the in-home sampling according to the following criteria: 1) adolescent living with adoptive or biological mother at the time of the interview, 2) adolescent had never been separated from mother for more than 6 months, 3) mother was in first marriage at the time of the interview, and 4) the adoptive mother had never been married to the adolescent's biological father. Of the 6577 adolescents in the final study sample, 214 (3.3%) were living with adoptive mothers and 6363 (96.7%) were living with biological mothers. Variables. The primary outcome measured was adolescent report of suicide attempt(s) in the past year. Other variables included in the analyses were sociodemographics characteristics (gender, age, race/ethnicity, family income, parental education), general health (self-rated health, routine examination in the past year, need for medical care in the past year that was not obtained), mental health (depressive symptoms, self-image, trouble relaxing in the past year, bad temper, psychological or emotional counseling in the past year), risk behaviors (cigarettes, alcohol, marijuana, sexual intercourse ever, delinquency, physical fighting in the past year, impulsive decision making), school-related characteristics (grade point average, school connectedness), and family interaction (family connectedness, parental presence, maternal satisfaction with parent-adolescent relationship). Data Analysis. Univariate analyses were used to compare adoptees versus nonadoptees, suicide attempters versus nonsuicide attempters, and adopted suicide attempters versus nonadopted suicide attempters on all variables. Variables that were associated with attempted suicide were entered into a forward stepwise logistic regression procedure, and variables that were associated with the log odds of attempt were retained in the model. The area under the model's receiver operating characteristic curve was calculated as a measure of its overall performance. After the association of adoption with attempted suicide was demonstrated, the potential mediating effect of impulsivity was explored by adding it to the model. The same procedure was followed for any variable that was associated with adoption in the full sample or the subsample of suicide attempters. To determine whether any variable in the model moderated the association between adoption and suicide attempt, the interaction term for that variable x adoption was forced into the model. RESULTS: Adoptees differed significantly from nonadoptees on 4 of 26 variables. They were more likely to have attempted suicide (7.6% vs 3.1%) and to have received psychological or emotional counseling in the past year (16.9% vs 8.2%), and their mothers reported higher parental education and family income. Attempters differed significantly from nonattempters on all variables except for age, race/ethnicity, parental education, family income, and routine examination in the past year. On logistic regression, 9 variables were independently associated with attempted suicide: depression (adjusted odds ratio [AOR]: 3.41), counseling (AOR: 2.83), female gender (AOR: 2.31), cigarette use (AOR: 2.31), delinquency (AOR: 2.17), adoption (AOR: 1.98), low self-image (AOR: 1.78), aggression (AOR: 1.48), and high family connectedness (AOR: 0.60). The receiver operating characteristic curve for the model had an area of 0.834, indicating performance significantly better than chance. The AOR for adoption did not change when parental education, family income, and impulsivity were forced into the model. (ABSTRACT TRUNCATED) PMID- 11483841 TI - Adolescents' perceptions of social status: development and evaluation of a new indicator. AB - OBJECTIVE: Eliminating health disparities, including those that are a result of socioeconomic status (SES), is one of the overarching goals of Healthy People 2010. This article reports on the development of a new, adolescent-specific measure of subjective social status (SSS) and on initial exploratory analyses of the relationship of SSS to adolescents' physical and psychological health. METHODS: A cross-sectional study of 10 843 adolescents and a subsample of 166 paired adolescent/mother dyads who participated in the Growing Up Today Study was conducted. The newly developed MacArthur Scale of Subjective Social Status (10 point scale) was used to measure SSS. Paternal education was the measure of SES. Indicators of psychological and physical health included depressive symptoms and obesity, respectively. Linear regression analyses determined the association of SSS to depressive symptoms, and logistic regression determined the association of SSS to overweight and obesity, controlling for sociodemographic factors and SES. RESULTS: Mean society ladder ranking, a subjective measure of SES, was 7.2 +/- 1.3. Mean community ladder ranking, a measure of perceived placement in the school community, was 7.6 +/- 1.7. Reliability of the instrument was excellent: the intraclass correlation coefficient was 0.73 for the society ladder and 0.79 for the community ladder. Adolescents had higher society ladder rankings than their mothers (micro(teen) = 7.2 +/- 1.3 vs micro(mom) = 6.8 +/- 1.2; P =.002). Older adolescents' perceptions of familial placement in society were more closely correlated with maternal subjective perceptions of placement than those of younger adolescents (Spearman's rho(teens <15 years) = 0.31 vs Spearman's rho(teens 15 years) = 0.45; P <.001 for both). SSS explained 9.9% of the variance in depressive symptoms and was independently associated with obesity (odds ratio(society) = 0.89, 95% confidence interval = 0.83, 0.95; odds ratio(community) = 0.91, 95% confidence interval = 0.87, 0.97). For both depressive symptoms and obesity, community ladder rankings were more strongly associated with health than were society ladder rankings in models that controlled for both domains of SSS. CONCLUSIONS: This new instrument can reliably measure SSS among adolescents. Social stratification as reflected by SSS is associated with adolescents' health. The findings suggest that as adolescents mature, SSS may undergo a developmental shift. Determining how these changes in SSS relate to health and how SSS functions prospectively with regard to health outcomes requires additional research. PMID- 11483842 TI - Syndromic ear anomalies and renal ultrasounds. AB - OBJECTIVE: Although many pediatricians pursue renal ultrasonography when patients are noted to have external ear malformations, there is much confusion over which specific ear malformations do and do not require imaging. The objective of this study was to delineate characteristics of a child with external ear malformations that suggest a greater risk of renal anomalies. We highlight several multiple congenital anomaly (MCA) syndromes that should be considered in a patient who has both ear and renal anomalies. METHODS: Charts of patients who had ear anomalies and were seen for clinical genetics evaluations between 1981 and 2000 at Cedars Sinai Medical Center in Los Angeles and Dartmouth-Hitchcock Medical Center in New Hampshire were reviewed retrospectively. Only patients who underwent renal ultrasound were included in the chart review. The literature was reviewed for the epidemiology of renal anomalies in the general population and in MCA syndromes with external ear anomalies. We defined a child as having an external ear anomaly when he or she had any of the following: preauricular pits and tags; microtia; anotia; or cup, lop, and other forms of dysplastic ears. A child was defined as having a renal anomaly if an ultrasound revealed any of the following: unilateral or bilateral renal agenesis; hypoplasia; crossed ectopia; horseshoe, pelvic, cystic kidney; hydronephrosis; duplicated ureters; megaureter; or vesicoureteric reflux. RESULTS: Because clinical genetics assessments were made by the same clinician at both sites (J.M.G.), data were combined. A total of 42 patients with ear anomalies received renal ultrasound; 12 (29%) of them displayed renal anomalies. Of the 12 patients with renal anomalies, 11 (92%) also received a diagnosis of MCA syndrome. Eleven of 33 patients (33%) with MCA syndromes had renal anomalies, whereas 1 of 9 patients (11%) with isolated ear anomalies had renal anomalies. Specific disorders seen were CHARGE association, Townes-Brocks syndrome, branchio-oto-renal syndrome, Nager syndrome, and diabetic embryopathy. CONCLUSIONS: We conclude that ear malformations are associated with an increased frequency of clinically significant structural renal anomalies compared with the general population. This is due to the observation that auricular malformations often are associated with specific MCA syndromes that have high incidences of renal anomalies. These include CHARGE association, Townes-Brocks syndrome, branchio-oto-renal syndrome, Nager syndrome, Miller syndrome, and diabetic embryopathy. Patients with auricular anomalies should be assessed carefully for accompanying dysmorphic features, including facial asymmetry; colobomas of the lid, iris, and retina; choanal atresia; jaw hypoplasia; branchial cysts or sinuses; cardiac murmurs; distal limb anomalies; and imperforate or anteriorly placed anus. If any of these features are present, then a renal ultrasound is useful not only in discovering renal anomalies but also in the diagnosis and management of MCA syndromes themselves. A renal ultrasound should be performed in patients with isolated preauricular pits, cup ears, or any other ear anomaly accompanied by 1 or more of the following: other malformations or dysmorphic features, a family history of deafness, auricular and/or renal malformations, or a maternal history of gestational diabetes. In the absence of these findings, renal ultrasonography is not indicated. PMID- 11483843 TI - Sensitization to common allergens and its association with allergic disorders at age 4 years: a whole population birth cohort study. AB - BACKGROUND: Atopy is defined as the genetic propensity to develop immunoglobulin E antibodies in response to exposure to allergens and assessed by skin prick test responses to common allergens. Although it is generally agreed that atopy is an important risk factor for allergic diseases such as asthma, rhinitis, and eczema, the extent to which atopy accounts for these diseases is controversial. OBJECTIVE: We aim to describe the prevalence of sensitization to common allergens and investigate the degree of association of atopy (as defined by positive skin prick test to 1 or more common allergens) to asthma, rhinitis, and eczema in a birth cohort at the age of 4 years. METHODS: A birth cohort of 1456 children was recruited over a 14-month period (1989-1990). These children have been seen previously at 1 and 2 years of age. At 4 years, 1218 children were reviewed and an interview was administered or postal questionnaire was completed for the presence of allergic diseases (asthma, rhinitis, and eczema). Additionally, in 981 children, skin prick tests with a battery of 12 common allergens were performed. Allergens were house dust mite (Dermatophagoides pteronyssimus), grass pollen mix, cat, dog, Alternaria alternata, Cladosporium herbarum, cow's milk, hen's egg, soya, cod, wheat, and peanut. A mean wheal diameter of at least 3 mm greater than the negative control was taken as positive. This analysis is confined to the 981 (67% of the original population) who also had skin prick tests to the standard battery. chi(2) tests were used to test the univariate association between each allergic disease and positive skin test. Multiple logistic regression analysis was performed to obtain the adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for the independent effect of sensitization to each allergen on allergic disease, adjusting for the effect of sensitization to other allergens. To ascertain how much of allergic disease is attributable to atopy, we estimated the population-attributable risk. This was calculated with the formula: P(R - 1) where R is the OR for the allergic disease under consideration and P is the proportion of atopy in children with that disease. RESULTS: Children who were skin prick-tested at 4 years were similar in most characteristics to the rest of the population, except that they had a higher prevalence of allergic disease. Allergic disorders (asthma, rhinitis, and eczema) were present in 276 (28.1%) of 981. One hundred ninety-two (19.6%) children were atopic (positive reaction to 1 or more allergens). Sensitization to inhalant allergens was relatively common (19.2%) as compared with food allergens (3.5%). House dust mite (11.9%), grass pollen (7.8%), and cat (5.8%) were the most common positive reactions. A test to the 4 most common allergens (house dust mite, grass pollen, cat, and A alternata) could detect 94% of the atopic children. Sensitization to the 4 most common allergens was strongly associated with the presence of allergic disorders. There was a graded effect with the potent allergens, such as house dust mite, having the greatest impact. For example, 50% of children sensitized to house dust mite had asthma as opposed to 44% sensitized to cat, 42% sensitized to grass pollen, and 32% sensitized to A alternata. Overall, 68.4% of children sensitized to house dust mite had asthma, eczema, and/or rhinitis. The respective figures for grass pollen, cat, and A alternata were 64.9%, 66.7%, and 57.4%. The proportion of children sensitized to cat was not higher in households with cat ownership (households with cats: 5.1% [19/374]; households without cats: 6.2% [36/580]; not significant [NS]). Similarly, no difference was seen in sensitization to dog in households with and without dogs (households with dogs: 1.8% [5/282]; households without dogs: 2.8% [19/673]; NS). Boys were atopic more often than girls at this age (male: 112 of 497 [22.5%] vs female: 80 of 484 [16.5%]; OR: 1.47, 95% CI: 1.07-2.02). Male preponderance was observed with most allergens, but this was statistically significant only for house dust mite (male: 75/497 [15.1%] vs female: 42/484 [8.7%]; OR: 1.87; CI: 1.25-2.79) and grass pollen (male: 51/497 [10.3%] vs female: 26/484 [5.4%]; OR: 2.01; CI: 1.23-3.29). An independent effect of allergen sensitization on asthma was observed only with house dust mite with an OR of 8.07 (CI: 4.60-14.14). The highest independent risk for rhinitis was sensitization to grass pollen (OR: 5.02; CI: 2.21-11.41), and for eczema, sensitization to peanut (OR: 4.65; CI: 1.02-21.34). The majority of children (98/192) were sensitized to >1 allergen. A graded effect was observed with the risk of allergic disease in the child increasing with the number of positive skin prick test reactions. This effect was consistent throughout the spectrum of allergic diseases (asthma, eczema, and rhinitis). (ABSTRACT TRUNCATED) PMID- 11483844 TI - Prenatal alcohol exposure and childhood behavior at age 6 to 7 years: I. dose response effect. AB - OBJECTIVE: Moderate to heavy levels of prenatal alcohol exposure have been associated with alterations in child behavior, but limited data are available on adverse effects after low levels of exposure. The objective of this study was to evaluate the dose-response effect of prenatal alcohol exposure for adverse child behavior outcomes at 6 to 7 years of age. METHODS: Beginning in 1986, women attending the urban university-based maternity clinic were routinely screened at their first prenatal visit for alcohol and drug use by trained research assistants from the Fetal Alcohol Research Center. All women reporting alcohol consumption at conception of at least 0.5 oz absolute alcohol/day and a 5% random sample of lower level drinkers and abstainers were invited to participate to be able to identify the associations between alcohol intake and child development. Maternal alcohol, cigarette, and illicit drug use were prospectively assessed during pregnancy and postnatally. The independent variable in this study, prenatal alcohol exposure, was computed as the average absolute alcohol intake (oz) per day across pregnancy. At each prenatal visit, mothers were interviewed about alcohol use during the previous 2 weeks. Quantities and types of alcohol consumed were converted to fluid ounces of absolute alcohol and averaged across visits to generate a summary measure of alcohol exposure throughout pregnancy. Alcohol was initially used as a dichotomous variable comparing children with no prenatal alcohol exposure to children with any exposure. To evaluate the effects of different levels of exposure, the average absolute alcohol intake was relatively arbitrarily categorized into no, low (>0 but <0.3 fl oz of absolute alcohol/day), and moderate/heavy (>/=0.3 fl oz of absolute alcohol/day) for the purpose of this study. Six years later, 665 families were contacted. Ninety-four percent agreed to testing. Exclusions included children who missed multiple test appointments, had major congenital malformations (other than fetal alcohol syndrome), possessed an IQ >2 standard deviations from the sample mean, or had incomplete data. The Achenbach Child Behavior Checklist (CBCL) was used to assess child behavior. The CBCL is a parent questionnaire applicable to children ages 4 to 16 years. It is widely used in the clinical assessment of children's behavior problems and has been extensively used in research. Eight syndrome scales are further grouped into Externalizing or undercontrolled (Aggressive and Delinquent) behavior and Internalizing or overcontrolled (Anxious/Depressed, Somatic Complaints, and Withdrawn) behaviors. Three syndromes (Social, Thought, and Attention Problems) fit neither group. Higher scores are associated with more problem behaviors. Research assistants who were trained and blinded to exposure status independently interviewed the child and caretaker. Data were collected on a broad range of control variables known to influence childhood behavior and/or to be associated with prenatal alcohol exposure. These included perinatal factors of maternal age, education, cigarette, cocaine, and other substances of abuse and the gestational age of the baby. Postnatal factors studied included maternal psychopathology, continuing alcohol and drug use, family structure, socioeconomic status, children's whole blood lead level, and exposure to violence. Data were collected only from black women as there was inadequate representation of other racial groups. STATISTICAL ANALYSES: Statistical analyses were performed using the SPSS statistical package. Frequency distribution, cross-tabulation, odds ratio, and chi(2) tests were used for analyzing categorical data. Continuous data were analyzed using t tests, analyses of variance (ANOVAs) with posthoc tests, and regression analysis. RESULTS: Testing was available for 501 parent-children dyads. Almost one fourth of the women denied alcohol use during pregnancy. Low levels of alcohol use were reported in 63.8% and moderate/heavy use in 13% of pregnancies. Increasing prenatal alcohol exposure was associated with lower birth weight and gestational age, higher lead levels, higher maternal age, and lower education level, prenatal exposure to cocaine and smoking, custody changes, lower socioeconomic status, and paternal drinking and drug use at the time of pregnancy. Children with any prenatal alcohol exposure were more likely to have higher CBCL scores on Externalizing (Aggressive and Delinquent) and Internalizing (Anxious/Depressed and Withdrawn) syndrome scales and the Total Problem Score. The odds ratio of scoring in the clinical range for Delinquent behavior was 3.2 (1.3-7.6) in children with any prenatal exposure to alcohol compared with nonexposed controls. The threshold dose was evaluated with the 3 prenatal alcohol exposure groups. One-way ANOVA revealed a significant between group difference for Externalizing (Aggressive and Delinquent) and the Total Problem Score. (ABSTRACT TRUNCATED) PMID- 11483845 TI - A new and improved population-based Canadian reference for birth weight for gestational age. AB - BACKGROUND: Existing fetal growth references all suffer from 1 or more major methodologic problems, including errors in reported gestational age, biologically implausible birth weight for gestational age, insufficient sample sizes at low gestational age, single-hospital or other non-population-based samples, and inadequate statistical modeling techniques. METHODS: We used the newly developed Canadian national linked file of singleton births and infant deaths for births between 1994 and 1996, for which gestational age is largely based on early ultrasound estimates. Assuming a normal distribution for birth weight at each gestational age, we used the expectation-maximization algorithm to exclude infants with gestational ages that were more consistent with 40-week births than with the observed gestational age. Distributions of birth weight at the corrected gestational ages were then statistically smoothed. RESULTS: The resulting male and female curves provide smooth and biologically plausible means, standard deviations, and percentile cutoffs for defining small- and large-for-gestational age births. Large-for-gestational age cutoffs (90th percentile) at low gestational ages are considerably lower than those of existing references, whereas small-for-gestational-age cutoffs (10th percentile) postterm are higher. For example, compared with the current World Health Organization reference from California (Williams et al, 1982) and a recently proposed US national reference (Alexander et al, 1996), the 90th percentiles for singleton males at 30 weeks are 1837 versus 2159 and 2710 g. The corresponding 10th percentiles at 42 weeks are 3233 versus 3086 and 2998 g. CONCLUSIONS: This new sex-specific, population-based reference should improve clinical assessment of growth in individual newborns, population-based surveillance of geographic and temporal trends in birth weight for gestational age, and evaluation of clinical or public health interventions to enhance fetal growth. fetal growth, birth weight, gestational age, preterm birth, postterm birth. PMID- 11483846 TI - Treatment of childhood asthma with anti-immunoglobulin E antibody (omalizumab). AB - BACKGROUND AND OBJECTIVE: There seems to be a strong causal relationship between allergy and the origins of asthma. Susceptibility to both is determined by a combination of genetics and environment acting through a complex network of cytokines. Nearly 90% of affected children have positive skin tests indicating the presence of specific immunoglobulin E (IgE), with sensitivity to house dust mite, Alternaria, cockroach, cat, and dog most closely linked to the disease. Greater exposure to house dust mite during infancy leads to earlier onset of wheezing, and elevated serum IgE levels correlate with the appearance of asthma symptoms. Specific IgE binds to high-affinity (FcepsilonRI) receptors on mast cells and basophils. The IgE-mediated reactions that follow exposure of sensitized mast cells to an allergen are designated early- and late-phase asthmatic responses (EAR and LAR). EAR is characterized by release of histamine and other preformed mediators within 1 hour of allergen exposure. It is often followed by LAR, an infiltration of the airways by inflammatory cells associated with an episode of more prolonged, and usually more severe airflow obstruction, 4 to 8 hours after antigen exposure. Chronic airway symptoms result from persistent LAR caused by continuous allergen exposure. IgE antibodies are capable of passive transfer of both EAR and LAR sensitivity. IgE-mediated mast cell activation contributes to chronic tissue eosinophilia and airway remodeling, with permanent loss in pulmonary function. Omalizumab (rhuMAb-E25) is a recombinant, humanized, monoclonal anti-IgE antibody of mouse origin developed for the treatment of IgE mediated diseases. Omalizumab binds to free IgE at the same site as the high affinity receptor. Although it attaches to free IgE, it does not bind to IgA, IgG, or cell-bound IgE. It therefore does not induce cross-linking of cell-bound IgE, which would lead to the release of allergic mediators. It has been reported to decrease serum IgE levels in a dose-dependent manner, inhibit EAR and LAR, and cause a down-regulation of FcepsilonRI receptors on basophils. Omalizumab has been reported to be safe and effective in improving asthma control and reducing the requirement for oral and inhaled corticosteroids. This double-blind, randomized, placebo-controlled study evaluated the safety, steroid-sparing effects, and impact on disease exacerbations of omalizumab in the treatment of childhood asthma. Methods. Participants were 334 males and premenarchal females aged 6 to 12 years, with moderate to severe allergic asthma requiring treatment with inhaled corticosteroids. During a run-in phase, all children were switched to equivalent doses of beclomethasone dipropionate (BDP), and the dose was adjusted to assure maintenance of asthma control achieved with previous corticosteroid treatment. Children were randomized to subcutaneously administered placebo (N = 109) or omalizumab (N = 225) at a dose based on body weight and initial serum IgE (0.016 mg/kg/IgE [IU/mL] per 4 weeks). BDP dose (initial range 168-420 microg/d) was kept stable for 16 weeks (stable-steroid phase), reduced over 8 weeks to the minimum effective dose (steroid-reduction phase), and maintained constant for the final 4 weeks. RESULTS: More participants in the omalizumab group decreased their BDP dose, and their reduction was greater than that of the placebo group (median reduction 100% vs 66.7%). BDP was withdrawn completely in 55% of the omalizumab group versus 39% of the placebo group. The incidence and the frequency of asthma exacerbations requiring treatment with doubling of BDP dose or systemic corticosteroids were lower in the omalizumab group. The treatment differences were statistically significant during the steroid-reduction phase, during which fewer participants in the omalizumab group had asthma exacerbation episodes (18.2% vs 38.5%), and the mean number of episodes per patient was smaller than with placebo (0.42 vs 2.72). Five asthma exacerbations requiring hospitalization all occurred in the placebo group. Participants' and investigators' global evaluations of treatment effectiveness were more favorable for omalizumab than placebo. Investigators rated effectiveness excellent for 31.5% of the omalizumab group versus 16.3% of the placebo group and good for 44.7% of the omalizumab group versus 32.7% of the placebo group. There was little change in asthma symptom scores or spirometry measurements during either the stable-steroid or steroid dose-reduction phase, with minimal differences between the treatment groups. The requirement for rescue medication in the omalizumab group during both the stable-steroid and steroid dose-reduction phases was consistently lower than at baseline. At week 28, the median number of puffs of rescue medication taken daily was 0 in the omalizumab group and 0.46 in the placebo group. The change from baseline was significant in favor of omalizumab. (ABSTRACT TRUNCATED) PMID- 11483847 TI - Intussusception, infection, and immunization: summary of a workshop on rotavirus. AB - This article summarizes the proceedings of a workshop sponsored by the National Institutes of Health and the National Vaccine Program Office, and held in Bethesda, Maryland, on January 21, 2000. The objective of the meeting was to focus research toward an understanding of the basis for the possible association between intussusception and the reassortant rhesus-human rotavirus vaccine tetravalent (RRV-TV). After numerous reports of intussusception after administration of RRV-TV, the manufacturers of this vaccine voluntarily withdrew it from the United States market. The American Academy of Pediatrics, the Advisory Committee on Immunization Practices, and the American Academy of Family Physicians also withdrew their original recommendations for administration of RRV TV to children at 2, 4, and 6 months of age. These actions will have global implications for the prevention of morbidity and mortality attributable to rotavirus infection. Benefit-cost ratios for the use of RRV-TV will be substantially different in developing countries compared with developed countries. Therefore, extensive research is needed in both of these settings, to further our understanding of the epidemiology, pathogenesis, and pathology of both rotavirus disease and intussusception to enable optimal prevention. The workshop reviewed the current understanding of the possible association between RRV-TV and intussusception, as well as the possible association between a variety of viral infections and intussusception. The workshop also identified critical areas of research regarding this possible association. This research will be essential not only for the development of safe and effective rotavirus vaccines, but for the development of other oral vaccines as well. PMID- 11483848 TI - Human immunodeficiency virus postexposure prophylaxis for adolescents and children. AB - Children and adolescents are at risk for human immunodeficiency virus (HIV) infection. Transmission occurs through perinatal exposures, injecting drug use, consensual and nonconsensual sex, needle-stick and sharp injuries, and possibly some unusual contacts. Youth engaging in high-risk sexual activities are especially endangered. Half of the estimated worldwide 5.3 million new HIV infections occur in adolescents and young adults aged 15 to 24. Of 20 000 known new adult and adolescent cases in the United States, 25% involve 13- to 21-year olds. More than 1.4 million children worldwide (aged 15 and younger) are believed to be infected, and >1640 new cases are diagnosed daily. Of the 432 000 people reported to be living with HIV or acquired immunodeficiency syndrome (AIDS) in the United States, 5575 are children under 13. HIV postexposure prophylaxis (PEP) is a form of secondary HIV prevention that may reduce the incidence of HIV infections. HIV PEP is commonly conceived of as 2 types: occupational and nonoccupational. Occupational HIV PEP is an accepted form of therapy for health care workers exposed to HIV through their jobs. A landmark study of healthcare workers concluded that occupational HIV PEP may be efficacious. Well-established US national guidelines for occupational HIV PEP exist for this at-risk population. Nonoccupational HIV PEP includes all other forms of HIV PEP, such as that given after sexual assault and consensual sex, injecting drug use, and needle-stick and sharp injuries in non-health care persons. Pediatric HIV PEP is typically the nonoccupational type. The efficacy of nonoccupational HIV PEP is unknown. The presumed efficacy is based on a collection of animal and human data concerning occupational, perinatal, and nonoccupational exposures to HIV. In contrast to occupational HIV PEP, there are no national US guidelines for nonoccupational HIV PEP, and few recommendations are available for its use for adolescents and children. Regardless of this absence, there is encouraging evidence supporting the value of HIV PEP in its various forms in pediatrics. Although unproven, the presumed mechanism for HIV PEP comes from animal and human work suggesting that shortly after an exposure to HIV, a window period exists during which the viral load is small enough to be controlled by the body's immune system. Antiretroviral medications given during this period may help to diminish or end viral replication, thereby reducing the viral inoculum to a more potentially manageable target for the host's defenses. HIV PEP is accepted practice in the perinatal setting and for health care workers with occupational injuries. The medical literature supports prescribing HIV PEP after community needle-stick and sharp injuries and after sexual assault from sources known or likely to be HIV-infected. HIV PEP after consensual unprotected intercourse between HIV sero-opposite partners has had growing use in the adult population, and can probably be utilized for children and adolescents. There is less documented experience and support for HIV PEP after consensual unprotected intercourse between partners of unknown HIV status, after prolonged or multiple episodes of sexual abuse from an assailant of unknown HIV status, after bites, and after the sharing of personal hygiene items or exposure to wounds of HIV infected individuals. There are no formal guidelines for HIV PEP in adolescents and children. A few groups have commented on its provision in pediatrics, and some preliminary studies have been released. Our article provides a discussion of the data available on HIV transmission and HIV PEP in pediatrics. In our article, we propose an HIV PEP approach for adolescents and children. We recommend a stratified regimen, based on the work of Gerberding and Katz and other authors, that attempts to match seroconversion risk with an appropriate number of medications, while taking into account adverse side-effects and the amount of information that is typically available upon initial presentation. Twice daily regimens should be used when possible, and may improve compliance. HIV PEP should be administered within 1 hour of exposure. We strongly recommend that physicians trained in this form of therapy review the indications for HIV PEP within 72 hours of its provision. We advocate that due diligence in determining level of risk and appropriateness of drug selection be conducted as soon as possible after an exposure has occurred. When such information is not immediately available, we recommend the rapid treatment using the maximum level of care followed by careful investigation and reconsideration in follow-up or whenever possible. HIV PEP may be initiated provisionally after an exposure and then discontinued if the exposure source is confirmed to not be HIV-infected. In most cases, consultations with the experts in HIV care can occur after the rapid start of therapy. (ABSTRACT TRUNCATED) PMID- 11483849 TI - Disseminated vaccine strain varicella as the acquired immunodeficiency syndrome defining illness in a previously undiagnosed child. AB - The Food and Drug Administration licensed a live-virus varicella vaccine (Varivax; Merck & Co Inc, West Point, PA) in March 1995. Prelicensure adverse events were minimal; however, since licensure and increased vaccine use, rare previously undetected risks have arisen. Presented here is the clinical course of a previously undiagnosed, human immunodeficiency virus-infected boy who developed dissemination of the vaccine strain of varicella zoster after immunization. chickenpox, human immunodeficiency virus, pneumonia, encephalopathy, varicella vaccine, adverse events, dissemination. PMID- 11483850 TI - Antibiotic treatment of pharyngitis. PMID- 11483851 TI - Cystic fibrosis. PMID- 11483852 TI - Ventricular and atrial septal defects. PMID- 11483853 TI - Index of suspicion. Case 1. Diagnosis: Recurrent or persistent rhinitis and bronchitis. PMID- 11483854 TI - Consultation with the specialist: renal tubular acidosis. PMID- 11483855 TI - Underphosphorylated BAD interacts with diverse antiapoptotic Bcl-2 family proteins to regulate apoptosis. AB - Survival factors activate kinases which, in turn, phosphorylate the proapoptotic Bcl-xl/Bcl-2-associated death promoter homolog (BAD) protein at key serine residues. Phosphorylated BAD interacts with 14-3-3 proteins, and overexpression of 14-3-3 attenuates BAD-mediated apoptosis. Although BAD is known to interact with Bcl-2, Bcl-w, and Bcl-xL, the exact relationship between BAD and anti- or proapoptotic Bcl-2 proteins has not been analyzed systematically. Using the yeast two-hybrid protein interaction assay, we found that BAD interacted negligibly with proapoptotic Bcl-2 proteins. Even though wild type BAD only interacted with selected numbers of antiapoptotic proteins, underphosphorylated mutant BAD interacted with all antiapoptotic Bcl-2 proteins tested (Bcl-2, Bcl-w, Bcl-xL, Bfl-1/A1, Mcl-1, Ced-9, and BHRF-1). Using nonphosphorylated recombinant BAD expressed in bacteria, direct interactions between BAD and diverse antiapoptotic Bcl-2 members were also observed. Furthermore, apoptosis induced by BAD was blocked by coexpression with Bcl-2, Bcl-w, and Bfl-1. Comparison of BAD orthologs from zebrafish to human indicated the conservation of a 14-3-3 binding site and the BH3 domain during evolution. Thus, highly conserved BAD interacts with diverse antiapoptotic Bcl-2 members to regulate apoptosis. PMID- 11483856 TI - Analysis of candidate antagonists of IAP-mediated caspase inhibition using yeast reconstituted with the mammalian Apaf-1-activated apoptosis mechanism. AB - We have reconstituted the Apaf-1-activated apoptosis mechanism in Sacchromyces cerevisiae such that the presence of a constitutively active form of Apaf-1 together with both Caspase-9 and Caspase-3 results in yeast death. This system is a good model of the Apaf-1-activated pathway in mammalian cells: MIHA (XIAP/hILP), and to a lesser degree MIHB (c-IAP1/HIAP2) and MIHC (c-IAP-2/HIAP1) can inhibit caspases in this system, and protection by IAPs (inhibitor of apoptosis) can be abrogated by coexpression of the Drosophila pro-apoptotic proteins HID and GRIM or the mammalian protein DIABLO/Smac. Using this system we demonstrate that unlike DIABLO/Smac, other proteins which interact with mammalian IAPs (TAB-1, Zap-1, Traf-1 and Traf-2) do not act to antagonise IAP- mediated caspase inhibition. PMID- 11483857 TI - Vascular endothelial cells express a functional fas-receptor due to lack of hemodynamic forces. AB - The fas system is present in atherosclerotic lesions. However, its role in the initiation and progression is still unclear. Here we show that in endothelial cells (EC) the expression of the fas receptor is regulated by flow conditions. The EC of the vascular system are regularly exposed to a range of hemodynamic forces with great impact on cellular structures and functions. Recently it was reported that in endothelial cells the lack of hemodynamic forces as well as irregular flow conditions trigger apoptosis by induction of a mechanosensitive autocrine loop of thrombospondin-1 and the alpha(V)beta(3) integrin/integrin associated protein complex. Here we show that EC cultivated under regular laminar flow conditions are devoid of the fas-receptor whereas cultivation under static conditions as well as under turbulence leads to its expression. Stimulation of the fas-receptor by its ligand increases the amount of apoptotic cells by twofold; the increase can be prevented by blocking the fas-receptor. The availability of the expressed fas receptor for stimulation by its ligand hints at a role as a tool for progression of atherosclerosis. PMID- 11483858 TI - Early decrease of survival signal-related proteins in spinal motor neurons of presymptomatic transgenic mice with a mutant SOD1 gene. AB - The mechanisms of motor neuronal death in amyotrophic lateral sclerosis (ALS) remain to be unclear. Phosphatidy-linositol 3-kinase (PI3-K) and its main downstream effector, Akt/protein kinase B (PKB) have been shown to play a central role in neuronal survival against apoptosis supported by neurotrophic factors. In order to investigate a possible impairment of survival signaling, we examined expressions of PI3-K and Akt in the spinal cord of the transgenic mice overexpressing a mutant Cu/Zn superoxide dismutase (SOD1) gene, a valuable model for human ALS. Immunoblotting and immunohistochemical analyses showed that the majority of spinal motor neurons lost the immunoreactivities for both PI3-K and Akt in the early and presymptomatic stage that preceded significant loss of the neurons. The present results suggest that an early decrease of survival signal proteins in the spinal motor neurons may account for the subsequent motor neuronal loss in this animal model of ALS. PMID- 11483859 TI - Down-regulation of protein kinase C activity by sorbitol rapidly induces apoptosis in human gastric cancer cell lines. AB - Although apoptosis-dependent involution of malignant tumors is associated with a number of non-surgical treatments including chemotherapy, most solid tumors, including gastric cancers, respond poorly to these therapies. In the hope of overcoming the resistance mechanism against non-surgical therapies, we studied the apoptosis-resistance mechanism in cells of gastric cancers as a model system. During the course of our study on apoptotic machinery in human gastric cancer cell lines, we previously found a rapid and efficient induction of apoptosis by sorbitol. In the present study, we demonstrated that the down-regulation of PKC activity in response to sorbitol is a major factor in the induction of apoptotic cell death in gastric cancer cells. PMID- 11483860 TI - Induction of cell death by adenoviruses. AB - Adenoviruses have proved to be excellent tools for gaining insight into the regulation, and deregulation, of the mammalian cell cycle. With the widespread clinical use of gene therapy fast approaching, there comes a need for a better understanding of how the cell death process is regulated. A greater understanding will allow the development of therapeutic approaches that both maximise transgene expression while minimising cytotoxicity to the target cell. Consequently, much adenovirus research has centered on understanding the mechanisms governing adenovirus induced cell death or apoptosis. This review discusses recent advances in the field of adenovirus cell death regulation and evaluates the roles of implicated gene products and their respective data. The data suggest the existence of multiple virus gene products involved in cell death regulation and point towards several distinct, yet related, cell death pathways. A discussion of the shortcomings of current adenoviral research, along with a proposed model based upon the data is also given. PMID- 11483861 TI - Caspase-independent apoptotic pathways in T lymphocytes: a minireview. AB - Cell death by apoptosis is involved in the maintenance of T cell receptor diversity, self tolerance, and T-cell number homeostasis. Until recently, apoptosis was thought to require caspase activation. Evidence is now accumulating that a caspase-independent pathway exists, shown by in vitro experiments with broad-range caspase inhibitors. Mature T lymphocytes readily undergo caspase independent apoptosis in vitro, and recent data suggest that this type of apoptosis may be involved in the negative selection of thymocytes. Mitochondria likely release death triggers specific for both caspase-dependent and caspase independent apoptotic pathways (cytochrome c and AIF respectively) in response to apoptotic stimuli. A caspase-independent pathway is triggered first in activated T lymphocytes subjected to apoptotic stimuli that do not rely on receptors with death domains. In this pathway, the early commitment phase to apoptosis involves cell shrinkage, peripheral DNA condensation and the translocation of mitochondrial AIF to the cytosol and nucleus. This process is reversible until mitochondrial cytochrome c is released and DeltaPsim dissipated. Only at this stage are caspases activated. PMID- 11483862 TI - Mechanism of fenretinide (4-HPR)-induced cell death. AB - 4-HPR (fenretinide) is a synthetic analog of retinoic acid (RA) whose potential as a chemopreventative agent has gained support from in vitro and animal experiments and in limited clinical trials. Comparative analyses of cellular, biochemical, and molecular properties of fenretinide with RA using various tissue culture cells reveal that a key distinction between these two retinoids lies in the ability of fenretinide to induce programmed cell death, also known as apoptosis. Here we review the composite evidence for induction of apoptosis in fenretinide-treated cells. Assays used to validate apoptosis in various cell types are also summarized. Apoptosis in response to fenretinide primarily occurs by a receptor-independent mechanism, which is accompanied by increases in signaling molecules, e.g., ceramide, and cysteine-dependent aspartate-directed proteases, termed caspases, including execution caspase-3. Both caspase-3 inhibitor DEVD-CHO and ceramide synthase inhibitor fumonisin B(1) (FB(1)) block fenretinide-induced apoptosis. Increase in caspase-3 appears to result from fenretinide-elicited stabilization of procaspase-3 zymogen. We also review apoptotic regulatory proteins such as inhibitor of apoptosis (IAPs) and second mitochondria-derived activator of caspase (SMACs) that participate in the coordinate control of caspase activities. The existence of a large number of proteins capable of modulating apoptosis via activation or inhibition of caspases, coupled with the fact that both the initiation and execution phases of apoptosis utilize pre-existing zymogens, which, once set in motion, culminates in an irreversible apoptotic cascade, raise the possibility that the on/off switch of apoptosis is linked to an intricate intracellular regulatory network, capable of responding to external stimuli such as fenretinide. This network functions to provide checks/balances of the need for apoptosis as well as to minimize and prevent untimely errors in apoptosis. We suggest that dynamic and coordinated regulation of apoptosis by such a hypothetical network in vivo may involve co localization of pro- and anti-apoptotic proteins and their respective activators/inhibitors in a macromolecular modular unit which we propose to be named caspasomes. Fenretinide also induces apoptosis by elevating reactive oxygen species (ROS), unrelated to changes in ceramide-caspases. Thus multiple, distinct pathways contribute to the induction of apoptosis by fenretinide. PMID- 11483863 TI - Cell death in the rat thymus: a minireview. AB - During the last decades, the literature has clearly established the fundamental role of the thymus in the development of an effective immune system. During thymocyte development and maturation, potentially autoreactive thymocytes are eliminated by a process known as apoptosis or programmed cell death responsible for the negative selection occurring within the thymus. This process is in sharp contrast to other types of cell death referred to as necrosis. Actually, three different types of cell death have been recently observed morphologically in the rat thymus, i.e. necrosis, apoptosis and clustered cell death. Moreover, among the numerous factors influencing thymocyte cell death, particular attention has been paid to hormones, chemicals, biological compounds and physical agents that may influence the type and/or the extent of cell death. Finally, a brief overview has been devoted to the contribution of mitochondria, nitric oxide, glutathione and intracellular levels of cations in addition to the activity of genes as cdk2, p53, Fas and members' of the Bcl2 family in modulating rat thymus cell death. PMID- 11483864 TI - Meeting report: mechanisms of cell death 2000. PMID- 11483865 TI - Action of an HMG CoA reductase inhibitor, lovastatin, on apoptosis of untransformed and ts-SV40 transformed human smooth muscle cells derived from saphenous vein. AB - The effect of lovastatin, an inhibitor of 3-hydroxymethyl-3-glutaryl coenzyme A (HMG CoA) reductase, was examined on human vascular smooth muscle cells (HVSMC). Untransformed HVSMC were obtained from saphenous vein and in addition an SV-40 transformed immortalized cell line (HVTs-SM1) derived from saphenous vein smooth muscle was also used. HVTs-SM1 cell proliferation and DNA synthesis were measured, and cell cycle analysis was performed by flow cytometry. Apoptosis in both cell types was assessed by a combination of flow cytometry, terminal deoxynucleotidyl transferase (TUNEL) reagent-based immunocytochemistry, DAPI staining, and DNA agarose gel electrophoresis. Lovastatin had no effect on apoptosis of HVSMC over 96 h in serum-free conditions or after stimulation with platelet-derived growth factor (PDGF-BB), although PDGF-BB increased apoptosis in HVSMC, and this was prevented by lovastatin. In HVTs-SM1 cells lovastatin inhibited cell proliferation and DNA synthesis and induced apoptosis in a time- and concentration-dependent manner. The effects of lovastatin on cell proliferation, DNA synthesis, and apoptosis were prevented by coincubation with mevalonate and geranylgeranyl pyrophosphate, but not by farnesyl pyrophosphate. Lovastatin does not induce apoptosis in saphenous vein HVSMC in culture and inhibits PDGF-BB-induced DNA synthesis and apoptosis. In contrast, in SV40 transformed immortalized HVTs-SM1 cells, lovastatin induces apoptosis and inhibits cell proliferation and DNA synthesis. The pro-apoptotic effects of lovastatin in SV40 transformed HVTs-SM1 cells may be related to the enhanced rate of proliferation or deregulation of the cell cycle in this cell line. PMID- 11483866 TI - Effect of cicletanine on the nitric oxide pathway in human umbilical vein endothelial cells. AB - The purpose of this study was to evaluate the effects of cicletanine, a slightly diuretic antihypertensive drug, on human vascular endothelial cells with regard to nitric oxide, intracellular calcium concentration ([Ca2+]i), cyclic nucleotide, inositol 1,4,5-trisphosphate (IP3), and prostacyclin generation. Primary cultured human umbilical vein endothelial cells were used in this study. [Ca2+]i was measured by fura-2/AM. Cyclic adenosine monophosphate (AMP), cyclic guanosine monophosphate (GMP), IP3, and prostacyclin were measured by radioimmunoassay. Nitric oxide was measured by the Griess method. Cicletanine had no effect on [Ca2+]i. Cicletanine (10(-6)-10(-4) M) increased cyclic GMP but decreased prostacyclin generation. Cicletanine had no stimulating effect on cyclic AMP or IP3 generation. IP3 increased 45Ca release from storage sites. Cicletanine decreased prostacyclin generation via increase in cyclic GMP. Cicletanine had no stimulating effect on nitrogen oxides for 2 h after incubation but increased it after 3-24 h. Pretreatment with L-N(G)-monomethyl-arginine (L NMMA) prevented this increase. The inhibitory effect of L-NMMA was prevented by pretreatment with L-arginine. These results indicate that nitric oxide and cyclic GMP may contribute to the antihypertensive action of cicletanine. PMID- 11483867 TI - Prohemorrhagic potential of dipyrone, ibuprofen, ketorolac, and aspirin: mechanisms associated with blood flow and erythrocyte deformability. AB - Dipyrone, ibuprofen, ketorolac, and aspirin were tested in a well-defined perfusion system (shear rates: 300/s, 800/s, and 1,800/s). Whole blood samples were treated with the drugs at analgesic doses and platelet interaction with damaged subendothelium was measured. All the drugs fully inhibited platelet cyclooxygenase, as assessed by classic aggregometry. Perfusion studies showed that there was a general tendency to reduce the percentage of large aggregates (thrombus; %T), to increase the percentage of adhered platelets (adhesion; %A), and to reduce the height of thrombi with respect to control. Aspirin significantly increased %A and reduced %T at all shear rates tested, whereas dipyrone had the same effect at 800/s, and ketorolac and ibuprofen at 1,800/s. In addition, aspirin significantly reduced erythrocyte deformability with respect to the other drugs. In conclusion, under our experimental conditions, aspirin showed the most remarkable effects on platelet function, closely followed by dipyrone. The effects of ketorolac were moderate, whereas ibuprofen had a minor impact on platelet function. PMID- 11483868 TI - Nebivolol induces calcium-independent signaling in endothelial cells by a possible beta-adrenergic pathway. AB - Nebivolol is a highly selective beta1-adrenoreceptor-blocking agent with a peculiar pharmacodynamic profile. It has peripheral acute vasodilating properties that are mediated by modulation of the endogenous production of nitric oxide. In this study we analyzed the different signaling pathways implicated in the response of human umbilical vein endothelial cells to nebivolol. Its effect on endothelial transduction pathways was determined by assaying phospholipase C and A2 activities and cyclic adenosine monophosphate (AMP) production. Variations in intracellular calcium concentration were also measured. Our results showed that nebivolol activates a calcium-independent transduction pathway that implicates an increase in adenylate cyclase and phospholipase A2 activity. Beta1- or beta2 Adrenoreceptor antagonists do not inhibit the action of nebivolol. However, its action on cyclic AMP production is inhibited by bupranolol, a beta1-3 adrenoreceptor antagonist, and S-(-)-cyanopindolol, a selective beta3 adrenoreceptor antagonist. Nebivolol also dose-dependently increased nitrite production. This effect was inhibited by bupranolol, suggesting that the possible action of nebivolol on beta3-adrenoreceptor is involved in its vasodilating properties. This study suggests that nebivolol could behave as a beta3 adrenoreceptor agonist and induce some calcium-independent pathways implicating phospholipase A2 and adenylate cyclase. This agonistic activity of nebivolol seems to be responsible for its endothelium-dependent vasodilating activity. PMID- 11483869 TI - Nicorandil enhances cardiac endothelial nitric oxide synthase expression via activation of adenosine triphosphate-sensitive K channel in rat. AB - In the heart, nitric oxide activates an adenosine triphosphate (ATP)-sensitive K (K(ATP)) channel that is constructed of two subunits, i.e., an ATP-binding cassette protein sulfonylurea receptor (SUR2) and a pore-forming inward rectifier (Kir6.1 or 6.2). However, whether this K(ATP) channel affects nitric oxide activation is unknown. Our aim was to assess whether pharmacologic activation of the K(ATP) channel by nicorandil contributes to endothelial nitric oxide synthase (eNOS) levels. A total of 21 7-week old male Sprague-Dawley rats were used. Seven were treated by intraperitoneal injection of nicorandil at 3 mg/kg/d; seven were treated with intraperitoneal nicorandil at 3 mg/kg/d after glibenclamide at 12 mg/kg/d twice a day p.o.; and seven were left untreated (controls). At 24 h after treatment, blood pressure and heart rate were measured, and eNOS, SUR2, Kir6.1, and Kir6.2 mRNA levels and eNOS protein levels in the left ventricle were determined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Nicorandil caused tachycardia without a change in blood pressure, whereas glibenclamide had no effect on the nicorandil-induced change in heart rate or on blood pressure. RT-PCR revealed that nicorandil increased the eNOS and SUR2 mRNA levels by 2.2- and 2.0-fold, respectively, (p < 0.01 versus control), and that these increases were completely inhibited by glibenclamide. A significant correlation was observed between eNOS and SUR2 mRNA levels in all experimental rats (r = 0.760, p < 0.001). However, Kir6.1 or 6.2 mRNA level was constant. Western blot analysis revealed that nicorandil caused a 1.6-fold increase in eNOS protein levels (p < 0.01 versus control). This increase was completely inhibited by glibenclamide. In conclusion, up-regulation of eNOS mRNA and protein levels by nicorandil, and inhibition of this upregulation by glibenclamide, were demonstrated in normotensive conscious rat hearts. Nicorandil appears to enhance cardiac eNOS expression via activation of a K(ATP) channel. PMID- 11483870 TI - Effects of volatile anesthetics on ryanodine-treated ferret cardiac muscle. AB - The volatile anesthetics halothane, isoflurane, and sevoflurane depress myocardial contractility by decreasing transsarcolemmal Ca2+ influx, Ca2+ release from the sarcoplasmic reticulum, Ca2+ sensitivity of the contractile proteins, and cross-bridge performance. The aim of this study is to assess and compare the effects of halothane, isoflurane, and sevoflurane on contractility in conditions in which sarcoplasmic reticulum Ca2+ release is abolished by pretreatment with ryanodine. Ferret right ventricular papillary muscles were exposed to ryanodine at 10(-6) M and then to incremental concentrations of halothane, isoflurane, or sevoflurane. In the presence of ryanodine, each anesthetic decreased isometric and isotonic contractility in a reversible, concentration-dependent manner with no differences between anesthetics and with little or no effect on time variables. It is likely that differences between anesthetic effects on contraction amplitude in isometric and isotonic twitches reside in their effects on the sarcoplasmic reticulum. PMID- 11483871 TI - Effects of red wine, alcohol, and quercetin on coronary resistance and conductance arteries. AB - Moderate consumption of alcoholic beverages is associated with a reduced risk of coronary heart disease (CHD). Some evidence suggests that red wine is particularly beneficial in this regard and may account in part for the French paradox, although the mechanism of this effect is unknown. We assessed the effects of red wine, ethanol, and quercetin, a major flavonoid constituent of red wine, in coronary resistance vessels (80-150 microm, i.d.) and conductance vessels (300-525 microm, i.d.) of the rabbit. Vessel wall tension was measured in isolated segments maintained in a wire-type myograph (37 degrees C) and preconstricted with 30 mM K+. At an alcohol concentration (14 mM) equivalent to moderate consumption, red wine evoked a small, transient constrictor effect in resistance and conductance vessels (9+/-4%, n = 5; 8+/-1%, n = 7, respectively; p < 0.05). Ethanol alone at this concentration was without effect. Quercetin (5.6, 8, and 30 microM) significantly relaxed resistance (-32+/-4%, n = 10; -47+/-2%, n = 7; -82+/-6%, n = 8, respectively) and conductance (-20+/-3%, n = 8; -32+/-4%, n = 8; -72+/-7%, n = 8, respectively) coronary arteries. Vasorelaxation by quercetin was endothelium-independent and was significantly greater in resistance than in conductance vessels. These data suggest that red wine and ethanol do not evoke relaxation in small coronary arteries at concentrations associated with moderate consumption. Quercetin elicits marked coronary vasorelaxation that is endothelium-independent. However, the concentrations of quercetin necessary to achieve this action are not attained with moderate red wine consumption. PMID- 11483872 TI - Formation of isoprostanes in children with type IIa hypercholesterolemia. AB - F2-isoprostanes are stable lipid peroxidation products of arachidonic acid and their quantification provides a novel approach to the assessment of oxidative stress in vivo. F2-isoprostanes are present in increased amounts in adult hypercholesterolemia, but no data exist concerning children. We investigated urinary isoprostaglandin F2, type III production as an index of lipid peroxidation in 15 children presenting with type IIa hypercholesterolemia (serum total cholesterol, 290 [SD +/- 70] mg/dl; low-density lipoprotein cholesterol, 210 [SD +/- 90] mg/dl) compared with 15 sex- and age-paired control children (serum total cholesterol, 160 [SD +/- 20] mg/dl). Urinary levels of isoprostaglandin F2alpha type III were measured by gas chromatography mass spectrometry. Urinary concentrations did not differ significantly in hypercholesterolemic children compared with control children (84.7 [SD +/- 37] vs. 96 [SD +/- 35] pmol/mmol creatinine, respectively). No significant correlation was found with total cholesterol, low-density-lipoprotein and high density-lipoprotein cholesterol, and apolipoprotein B and A1 serum levels. F2 isoprostane urinary levels in children with type IIa hypercholesterolemia do not differ from those of age- and sex-matched control children and are not correlated to blood lipid parameters, suggesting that hypercholesterolemia is not associated with increased lipid peroxidation in childhood. PMID- 11483873 TI - Kappa-opioid receptor agonist inhibits the cholera toxin-sensitive G protein in the heart. AB - To explore the signaling mechanisms of the negative modulation of beta adrenoceptors by kappa-Opioid receptors (kappa-OR) in the heart, the possibility of the interaction at the level of G protein and receptor was determined. Cholera toxin, an activator of the stimulatory G protein (Gs), elevated electrically induced intracellular Ca2+ ([Ca2+]i) transients and induced ribosylation of the alpha-subunit of Gs (Gsalpha) in rat ventricular myocytes. The effects were significantly attenuated by U50,488H, a specific agonist of kappa-OR, and were abolished by nor-binaltorphimine, a selective kappa-OR antagonist. The content of Gsalpha, however, was not affected by U50,488H. Receptor binding experiments showed that neither Bmax nor Kd of the binding of [3H]CGP-12177, a beta adrenoceptor antagonist, was affected by U50,488H. The current study provides the first evidence that kappa-OR stimulation inhibits the ribosylation of the alpha subunit of the Gs protein, thus inhibiting the action of cholera toxin on the protein. PMID- 11483874 TI - 17 Beta-estradiol increases nitric oxide and prostaglandin I2 production by cultured human uterine arteries only in histologically normal specimens. AB - These experiments were designed to investigate whether 17beta-estradiol (E2) modulates the endothelial function of perimenopausal human uterine arteries. After the artery specimen was cultured in the presence or absence of E2 at a physiologic concentration of 200 pg/ml, changes in isometric tension and cyclic nucleotide production were determined. Degree of intimal hyperplasia was assessed histologically and expressed as intima-to-media ratio. Acetylcholine produced an endothelium-dependent relaxation in six specimens (group I) of 12, which was inhibited by NG-nitro-L-arginine or indomethacin. However, the agonist failed to produce a definite relaxation in the remaining 6 (group II). The endothelium dependent relaxation was significantly augmented after incubating with E2 only in group I specimens. Cyclic nucleotide production was significantly increased after E2 incubation only in group I specimens, whereas it was inhibited by NG-nitro-L arginine or indomethacin. Histologic study revealed that the six specimens of group I had normal intima (intima-to-media ratio = 19.1+/-1.8%) and the remaining six of group II had intimal hyperplasia (intima-to-media ratio = 53.6+/-5.3%). Increased production of cyclic nucleotides occurred in uterine arteries with normal intima but not in arteries with intimal hyperplasia derived from perimenopausal women. PMID- 11483875 TI - Improvement in endothelial dysfunction in patients with hypoalphalipoproteinemia and coronary artery disease treated with bezafibrate. AB - Isolated low high-density lipoprotein cholesterol (HDLc) is a well-known risk factor for cardiovascular disease and is associated with arterial endothelium dysfunction. Several studies have shown that cholesterol lowering in patients with hypercholesterolemia improves endothelial function, but the effect of treating low HDLc levels remains unknown. We studied the effect of increasing HDLc on endothelial function in patients with coronary artery disease (CAD) and isolated low HDLc (HDLc) <0.91 mM, low-density lipoprotein cholesterol (LDLc) <4.1 mM, and triglycerides <2.8 mM. Flow-mediated endothelium-dependent dilatation (FMD) in response to reactive hyperemia was measured by brachial ultrasound, before and after bezafibrate treatment (400 mg daily for 6 months) in 16 patients with CAD and impaired FMD (<10%). After bezafibrate therapy, HDLc increased from 0.79-1.0 mM (p = 0.0008) at the expense of both HDL2 and HDL3 subfractions, apolipoprotein A-I increased from 1.04-1.19 g/l (p = 0.0012), and fibrinogen decreased from 4.45-3.39 g/l (p = 0.0007). The impaired FMD increased after bezafibrate treatment from a median of 2.5-12.3% (p = 0.0004). Endothelial function was normalized in eight patients (50%), improved in four (25%), and did not change in four (25%). These observations indicate that in patients with isolated low HDLc and CAD, bezafibrate treatment improves endothelial function of brachial arteries, increases HDLc and apolipoprotein A-I, and lowers fibrinogen concentrations. PMID- 11483876 TI - Myocardial expression of the endothelin system in endotoxin-treated rats. AB - Although circulating plasma levels of endothelin (ET)-1 are elevated in endotoxemia, little is known about the myocardial expression of the ET system in endotoxic shock. We assessed the temporal mRNA expression pattern of key components of the ET system (pre-pro ET (ppET) -1, -2, ET-converting enzyme-1, ET(A) and ET(B) receptors) by reverse transcription polymerase chain reaction in a rat model of early endotoxic shock. Lipopolysaccharide (5 mg/kg, i.p.) caused a transient increase (p < 0.05) in inducible nitric oxide synthase mRNA expression. ppET-1 mRNA expression was increased at 2 h (approximately 12-fold increase; p < 0.05) in the lipopolysaccharide compared with the saline group and ppET-2 mRNA expression was unaltered. ET-converting enzyme-1, ET(A), and ET(B) receptor mRNA expression was unaltered in the lipopolysaccharide compared with the saline group. While ppET-1 mRNA expression is selectively upregulated in ventricular myocardium of lipopolysaccharide-treated rats, an absence of alteration in ET converting enzyme-1 mRNA expression suggests an excess capacity of ET-converting enzyme-1 to cope with the increased expression of ET-1. At the level of the receptor, endotoxic shock did not affect the expression of either ET(A) or ET(B) receptor mRNA. These data are consistent with the increased expression of myocardial ET-1 as an acute-phase response due to hemodynamic instability associated with the early stages of endotoxic shock. PMID- 11483877 TI - Electropharmacologic effects of a new phosphodiesterase III inhibitor, toborinone (OPC-18790), assessed in an in vivo canine model. AB - Electropharmacologic effects of a new phosphodiesterase (PDE) III inhibitor toborinone (OPC-18790) were assessed in a halothane-anesthetized, closed-chest canine model. Toborinone, 0.03 mg/kg, increased ventricular contractility, decreased total peripheral resistance, and inhibited intraventricular conduction without changing other cardiovascular parameters. A clinically relevant dose of 0.3 mg/kg increased heart rate, systolic blood pressure, and cardiac output, decreased preload to the left ventricle, enhanced atrioventricular nodal conduction, and shortened repolarization and the vulnerable period of the ventricle, in addition to enhancing the effects observed after the low dose. A high dose of 3 mg/kg of toborinone decreased systolic, mean, and diastolic pressures and prolonged the effective refractory period (ERP) in addition to the effects observed after the middle dose. No further change was detected in ventricular repolarization. Most of the cardiohemodynamic effects can be explained by the PDE III inhibition by toborinone. With regard to electrophysiologic properties, the prolongation of intraventricular conduction time and ERP by toborinone suggests sodium channel inhibition. The lack of the prolongation of ventricular repolarization suggests that previously demonstrated inhibition of I(Kr) and I(K1) and increased I(Ca-L) by toborinone might be counteracted by factors such as the cyclic AMP-dependent outward currents, I(Ks) and I(C1). PMID- 11483878 TI - Delayed or second window preconditioning induced by adenosine A1 receptor activation is independent of early generation of nitric oxide or late induction of inducible nitric oxide synthase. AB - Transient adenosine A1 receptor (A1R) activation induces a second window or delayed preconditioning against myocardial infarction 24-72 h later. Early generation of nitric oxide and delayed induction of nitric oxide synthase have been implicated in mediating delayed cardioprotection after ischemic preconditioning in rabbits. Recent evidence indicates that some of the regulatory roles of adenosine in cardiac tissue may be mediated by A1R-induced generation of nitric oxide. This study examined the role of nitric oxide in the mediation of A1R-induced delayed preconditioning against infarction. Pharmacologic preconditioning of rabbits with the selective A1R agonist 2-chloro-N6 cyclopentyladenosine 100 microg/kg (CCPA) significantly reduced myocardial infarct size compared with control animals, after 30 min regional ischemia and 2 h reperfusion in vivo 24 h later (27.3+/-4.7 vs. 46.0+/-3.7%, respectively; p = 0.001). Nonselective inhibition of nitric oxide synthase with N(G)-nitro-L arginine methyl ester (10 mg/kg) before administration of CCPA did not affect this infarct limitation at 24 h. Selective inhibition of inducible nitric oxide synthase before the prolonged ischemic insult on day 2, with two structurally independent inducible nitric oxide synthase inhibitors, L-N(6)-(1-iminoethyl) lysine (10 mg/kg) or aminoguanidine (300 mg/kg), did not abrogate the reduction in infarction observed by pharmacologic preconditioning with CCPA 24 h earlier. These results suggest that the second window or delayed protection against myocardial infarction observed 24 h after pharmacologic preconditioning with an adenosine A1 agonist occurs independently of either early generation of nitric oxide or subacute induction of inducible nitric oxide synthase. PMID- 11483879 TI - Role of short-term inhibition of factor Xa by FXV673 in arterial passivation: a study in a chronic model of thrombosis in conscious dogs. AB - Factor Xa (fXa) plays a pivotal role in the activation of the coagulation system during thrombosis, but, unlike GPIIb/IIIa receptor antagonists, the role of fXa inhibition in arterial passivation is not well defined. We compared the long-term antithrombotic efficacy of a direct fXa inhibitor, FXV673, and heparin after short-term infusion in conscious dogs. Dogs were instrumented surgically to induce carotid artery thrombosis by electrolytic injury. On day 1, dogs received a 3-h infusion of placebo (n = 10), FXV673 (100 microg/kg + 10 microg/kg/min, n = 7), or heparin (60 U/kg + 0.7 U/kg/min, n 7). Injury (100 microA) was initiated concomitantly for 1 h. The procedure was repeated on day 2 with injury of 200 microA for 3 h. Carotid artery blood flow (CBF) and coagulation parameters were monitored continuously for 3 h on days 1 and 2 and for 30 min on days 3, 4, and 5. On day 1 at 3 h, CBF in the placebo-treated group was 26% of baseline with 70% incidence of occlusion. None of the vessels occluded in the heparin and FXV673 groups; however, the CBF was significantly higher in the FXV673 group (92+/-8 ml/min versus 39+/-12 ml/min). Before injury on day 2, CBF recovered in all groups to 71-89% of baseline. After the second injury, all vessels in the placebo treated group progressed to complete occlusion by 3 h. CBF was significantly higher in FXV673 group compared with heparin throughout the 3-h period. On days 3, 4, and 5 the placebo-treated vessels remained occluded, but the CBF in the heparin group was 33+/-20 ml/min, 55+/-11 ml/min and 68+/-12 ml/min, respectively, compared with 84+/-10 ml/min, 98+/-7 ml/min, and 99+/-10 ml/min in the FXV673 group. The arterial thrombus mass was significantly lower in FXV673 group (13+/-4 mg) compared with placebo (103+/-10 mg) and heparin (44+/-11 mg). In summary, these data demonstrate that short-term infusion of FXV673 was associated with long-term efficacy that was superior to standard heparin and underscore the role of direct fXa inhibition in arterial passivation. PMID- 11483880 TI - Pharmacologic modulation of the immune interaction between cytotoxic lymphocytes and ventricular myocytes. AB - Numerous studies have demonstrated that immune effector mechanisms cause serious heart diseases, among which are heart transplant rejection, myocarditis, and the resulting dilated cardiomyopathy, as well as Chagas' disease. Whereas different effectors of the immune system can affect cardiac function, this review primarily focuses on the immune damage caused by cytotoxic T lymphocytes. The immune attack staged by cytotoxic T lymphocytes is carried out by one of two distinct modes of lymphocytotoxicity: (a) secretion of lytic granules containing the pore-forming protein perforin and a family of serine proteases (i.e., granzymes) and (b) interaction between the lymphocyte Fas ligand and the target cell Fas receptor. Ventricular myocytes challenged by the immune system sustain diverse intracellular changes, among which the rise in intracellular calcium ([Ca2+]i) constitutes an important contributor to myocyte dysfunction. Hence, this [Ca2+]i rise, which does not necessarily result in apoptosis, can affect cardiac function directly and indirectly. Importantly, the final outcomes of these perturbations vary markedly and depend on intracellular circumstances such as the magnitude of the absolute rise in [Ca2+]i and its temporal and spatial determinants, the metabolic status of the myocyte, as well as a fine balance between pro-apoptotic and anti-apoptotic factors. In view of the central role of [Ca2+]i rise in immune mediated myocyte dysfunction and possibly cell death, this review addresses three topics related to the immune assault on the heart: (a) [Ca2+]i rise in affected myocytes; (b) the source for the [Ca2+]i rise; and (c) pharmacologic modification of the immune-mediated [Ca2+]i rise. PMID- 11483881 TI - Endothelin mediates the altered renal hemodynamics associated with experimental congestive heart failure. AB - Congestive heart failure (CHF) is commonly associated with renal dysfunction. The goal of the current study was to evaluate the role of endothelin in the renal dysfunction of experimental CHF by using tezosentan, a potent dual endothelin receptor antagonist. Rats were subjected to coronary artery ligation. Cardiac and renal hemodynamics were assessed after 3-5 weeks, when CHF had developed. Compared with control rats, CHF rats had significantly higher left ventricular end-diastolic pressure (LVEDP), lower mean arterial pressure, and reduced dP/dt(max). CHF rats had severe renal vasoconstriction, as assessed by increased renal vascular resistance (RVR, p < 0.001), decreased renal plasma flow (RPF, p < 0.001), and glomerular filtration rate (GFR, p < 0.001). Filtration fraction rose (p < 0.001). Urine flow rate and sodium excretion were markedly lower. Acute administration of tezosentan induced a marked decrease in LVEDP without change of dP/dt(max) and heart rate. Tezosentan decreased RVR (-43%, p < 0.001) and increased RPF and GFR. Filtration fraction decreased slightly. Tezosentan also increased urine flow rate and sodium excretion. These findings demonstrate that endothelin at least partly mediates the altered renal hemodynamics associated with experimental CHF. Dual endothelin receptor blockade could be useful for the improvement of both cardiac and renal function in CHF. PMID- 11483882 TI - Hypoxic vasoconstriction of rat main pulmonary artery: role of endogenous nitric oxide, potassium channels, and phosphodiesterase inhibition. AB - This study investigated the influence of NO, potassium (K+) channel blockade, and the phosphodiesterase inhibitors (PDEIs) theophylline (non-selective PDEI), siguazodan (PDE3I), rolipram (PDE4I), and zaprinast (PDE5I) on rat isolated main pulmonary artery hypoxic (95% N2 and 5% CO2) vasoconstriction. Hypoxic vasoconstriction increased by 27% (p < 0.01) in the presence of the NO synthase inhibitor L-NAME (10(-4) M), and by 15% (p < 0.05) in the presence of the K(ATP) channel blocker glibenclamide (10(-6) M), without potentiation by the combination of these two drugs. Hypoxic vasoconstriction decreased by 28% (p < 0.01) in presence of the Kv,-voltage-dependent channel blocker 4-aminopyridine (10(-3) M), whereas the other K+ channel blockers, charybdotoxin (BKCa, large-conductance Ca2+-sensitive K+ channels) and apamin (SKCa, small-conductance Ca2+-sensitive K+ channels) had no effect. The nonselective PDEI theophylline induced a concentration-dependent relaxation (pD2 = 4.05, Emax = 90% [expressed as a percentage of maximal relaxation induced by papaverine 10(-4) M]). Among the selective PDEIs, siguazodan was significantly (p < 0.01) more efficient than rolipram and zaprinast (Emax values were 84%, 67%, and 58%, respectively) and significantly (p < 0.05) more potent than zaprinast (pD2 values were 6.48, 6.34, and 6.16 for siguazodan, rolipram, and zaprinast). Glibenclamide and L-NAME significantly (p < 0.05) shifted the concentration-response curve (CRC) for zaprinast to the right, and L-NAME shifted the CRC significantly to the right for siguazodan. In the presence of L-NAME, glibenclamide had no effect on the CRC of zaprinast. We conclude that (a) NO exerts a permanent inhibitory effect against hypoxic vasoconstriction that might be mediated in part by an activation of K(ATP) channels; (b) a 4-aminopyridine-sensitive K+ channel is involved in vasoconstriction under hypoxic conditions; (c) PDEs 3 and 5 are the predominant PDE isoforms in rat pulmonary artery relaxation; and (d) NO and K(ATP), but neither BK(Ca), SK(Ca), nor Kv channels, are involved in the relaxant effect of PDEIs. PMID- 11483884 TI - Mediastinal staging of non-small-cell lung cancer. AB - BACKGROUND: The goal of preoperative staging of non-small-cell lung cancer (NSCLC) is to identify patients who will benefit from surgical resection. Various imaging and less invasive modalities are now available to improve therapy decision making. METHODS: The available staging methods are reviewed, including conventional methods, surgical staging, and less invasive means of pathologic staging. RESULTS: Computed tomography alone is not sufficiently accurate to stage the mediastinum, and further definitive testing is usually indicated. Positron emission tomography, along with mediastinal biopsy techniques using transbronchial needle aspiration or endoscopic ultrasound, has the potential to improve the accuracy of pretreatment staging. CONCLUSIONS: Every effort should be made to accurately discriminate between benign and malignant mediastinal disease. With further research on the proper roles of these new imaging modalities, they will become more widely used and will improve the accuracy of pretreatment staging of NSCLC. PMID- 11483885 TI - Treatment of stage I and II non-small-cell lung cancer. AB - BACKGROUND: The appropriate staging and treatment of patients with stage I and II non-small-cell lung cancer (NSCLC) are important in that the potential for a lost curative opportunity in this population is greater than for those presenting with advanced NSCLC. METHODS: Treatment options -- surgery, radiation therapy, and chemotherapy -- for stage I and II NSCLC are reviewed, and the impact of newer staging modalities on patient survival is discussed, including altering both the lead-time and clinicopathologic stage biases that exist in the diagnosis and treatment of NSCLC. Some predictions are also made regarding how that standard may change for clinicians in the near future, and methods for further improvements in posttreatment survival in this group are discussed. RESULTS: Whenever possible, patients with early-stage NSCLC should be treated with surgical resection. Patients for whom resection is not an option may benefit from radiation as definitive therapy. Positive results with neoadjuvant chemotherapy have led to an ongoing randomized trial (Intergroup S9900) to compare surgery alone to neoadjuvant chemotherapy plus surgery. CONCLUSIONS: Staging bias may affect the overall low survival of early-stage NSCLC. However, true stage specific survival may improve with newer imaging modalities. Future advances, including closed transthoracic radiation, thermal ablative therapy techniques, and gene therapy, may supplant the need to surgically resect these tumors to achieve local control. PMID- 11483886 TI - Treatment of advanced non-small-cell lung cancer: a review of current randomized clinical trials and an examination of emerging therapies. AB - BACKGROUND: Lung cancer continues to be the leading cause of cancer-related deaths for Americans. As most patients present with nonsurgically curable disease, major efforts have been made in the treatment of advanced non-small-cell lung cancer (NSCLC) with chemotherapy. Several new agents and new combinations of chemotherapy are available. METHODS: The author reviews randomized clinical trials investigating chemotherapy for advanced NSCLC in chemotherapy-naive patients, in patients who present with relapsed or progressive disease, and in elderly patients. Therapies that incorporate new biological agents to target specific aberrations in lung cancer are discussed. RESULTS: Several clinical trials demonstrate improvement in overall survival as well as quality of life with chemotherapy treatment of advanced NSCLC. Better options are available for patients who have relapsed after first-line chemotherapy, and treatment of elderly patients with chemotherapy has demonstrated benefit in survival and quality of life. New agents that target molecular pathways are being tested in patients with early-stage disease. CONCLUSIONS: Despite progress with newer agents for the treatment of advanced NSCLC, only 14% of patients with the disease are alive at 5 years after initial diagnosis. New therapies are needed. PMID- 11483887 TI - Endobronchial management of advanced lung cancer. AB - BACKGROUND: Patients with lung cancer often have bulky endobronchial disease, endobronchial extension, or airway compression. Many endobronchial treatment modalities are available to supplement traditional therapies for advanced lung cancer. METHODS: The author reviews the use of several endobronchial treatment modalities that can augment standard antitumor therapies for advanced lung cancer, including rigid and flexible bronchoscopy, laser therapy, endobronchial prosthesis, and photodynamic therapy. RESULTS: Since the early 1980s, technical advances in interventional techniques have enhanced symptom-free survival and quality of life for patients with lung cancer. Although interventional procedures are not definitive therapies, they often relieve the strangling sensation produced by airway occlusion. CONCLUSIONS: Endobronchial interventions are important adjuncts in the multimodality management of lung cancer and should become standard considerations in the management of patients with advanced lung cancer. For patients with respiratory symptoms associated with their disease, these interventions provide symptom palliation and improved quality of life. PMID- 11483888 TI - Modular endoprostheses for children with malignant bone tumors. AB - BACKGROUND: The treatment of malignant bone tumors in skeletally immature patients is difficult because the required surgery removes the growth plate of one extremity. To date, few techniques for limb salvage have been successful due to complications involving limb expansion and limb-length discrepancy. Newer technology is now available for prosthetic replacements for modular expansion. METHODS: The authors review current surgical treatment of malignant bone tumors in children, with emphasis on reconstruction with expandable modular prostheses, and they present their own experience. RESULTS: Thirty-seven children with malignant bone tumors underwent primary tumor resection and reconstruction with a modular prosthetic device. Fourteen had subsequent successful expansions with modular prostheses. A new prosthesis, in which lengthening is achieved by an external electromagnetic field rather than an open surgical procedure, is discussed. CONCLUSIONS: Limb-preserving surgery in children with malignant bone tumors can be accomplished successfully with modern prosthetic devices that have expandable modules within them. The technique of expandable prostheses using electromagnetic fields rather than surgical interventions shows promise. PMID- 11483889 TI - Histologic variants of squamous cell carcinoma of the skin. PMID- 11483890 TI - Facial cellulitis by Pseudomonas aeruginosa in a neutropenic patient. PMID- 11483891 TI - Spirituality and medicine: a proposal. PMID- 11483892 TI - Synthesis and evaluation of novel aldose reductase inhibitors: Effects on lens protein kinase Cgamma. AB - PURPOSE: To synthesize novel aldose reductase inhibitors (ARI) that will normalize losses in protein kinase Cgamma (PKCgamma) observed during diabetes and galactosemia. METHODS: ARI were synthesized as tricyclic pyrones 1-6 (HAR-1 through HAR-6) from 3-methyl-1H,7H-5a,6,8,9-tetrahydro-1-oxopyrano[4,3 b][1]benzopyran and (5aS,7S)-7-isopropenyl-3-methyl-1H,7H-5a,6,8,9-tetrahydro-1 oxopyrano[4,3-b][1]benzopyran and were tested by inhibition of aldose reductase enzyme activity in vitro and by inhibition of polyol formation in lens epithelial cells in culture. Identified compounds were further tested in galactosemic rat lens in vivo for (a) normalized PKCgamma levels by Western blot, (b) reduction of phosphorylation of the gap junction protein Cx46 by analyses of co immunoprecipitated proteins, and (c) by normalization of gap junction activity as measured by dye transfer. RESULTS: HAR-1 (1H,7H-5a,6,8,9-tetrahydro-1 oxopyrano[4,3-b][1]benzopyran-3-acetic acid) was identified as an ARI with IC50 for aldose reductase inhibition at 2 nM. Polyol accumulation in lens epithelial cells was reduced by 80% at 10 microM. Rats fed 40% galactose for 9 days had an 80% reduction in PKCgamma levels which were normalized by HAR-1 at 100 mg/kg/day, fed orally. Phosphorylation of Cx46 was increased by 50% and this was normalized in HAR-1 treated rats (6 day treatment). Gap junction activity of galactosemic rats was reduced by 55% and this was normalized by HAR-1 in six day-treated rats. CONCLUSIONS: HAR-1 is a novel ARI which normalized losses of PKCgamma, changes in Cx46 phosphorylation, and gap junction activity. PMID- 11483893 TI - Analysis of alpha-crystallin chaperone function using restriction enzymes and citrate synthase. AB - PURPOSE: To compare the abilities of [alpha]A-crystallin, [alpha]B-crystallin, and mini-[alpha]A-crystallin (a synthetic peptide chaperone representing the functional unit of [alpha]A-crystallin) to protect against heat-induced inactivation of citrate synthase (CS) and restriction enzymes, SmaI and NdeI. METHODS: Restriction enzymes, SmaI and NdeI were heated at different temperatures in the presence of various amounts of molecular chaperones and tested for their ability to cleave plasmid DNA. The aggregation of CS was measured at 43 degrees C while the loss in activity was monitored at 37 degrees C in the presence of various crystallins. RESULTS: Restriction enzyme activities were protected by the crystallin subunits up to 37 degrees C for SmaI and 43 degrees C for NdeI. However, the mini-[alpha]A-crystallin was unable to protect endonuclease activity. The crystallin subunits and the peptide chaperone were able to suppress thermal aggregation of CS at 43 degrees C, but failed to stabilize its activity at 37 degrees C. CONCLUSIONS: The ability of [alpha]-crystallin subunits to stabilize denaturing proteins varies from enzyme to enzyme as evidenced by the inactivation of CS and protection of SmaI and NdeI activity in the presence of [alpha]-crystallin subunits. Additionally, our results show that there could be more than one site in [alpha]A-crystallin responsible for its chaperone-like action. By addition of crystallin subunits to restriction enzymes prior to or during storage, transport, or assay would maintain or improve their activity thereby decreasing their cost. PMID- 11483894 TI - Spectroscopic analysis of lens recombinant betaB2- and gammaC-crystallin. AB - PURPOSE: To compare the spectroscopic and unfolding properties of human lens [beta]B2- and [gamma]C-crystallin with those of [alpha]A-crystallin. METHODS: Human lens [beta]B2- and [gamma]C-crystallin were cloned and measured spectroscopically. The unfolding curves in response to guanidine HCl (GdnHCl) and heat were also obtained by measuring Trp fluorescence emission intensity or emission maximum wavelength with increasing perturbation. RESULTS: Very similar spectroscopic and unfolding properties were seen with [beta]B2- and [gamma]C crystallin, but both demonstrated great differences compared with [alpha]A crystallin. Unlike [alpha]A-crystallin, [beta]B2- and [gamma]C-crystallin showed very little binding to Bis-ANS (4,4'-dianilino-1,1'-binaphthalene-5,5'-disulfonic acid), a hydrophobic fluorescence probe. Both [beta]B2- and [gamma]C-crystallin were more resistant than [alpha]A-crystallin to GdnHCl-induced unfolding, but [alpha]A-crystallin was more resistant than [beta]B2- and [gamma]C-crystallin to heat induced unfolding. CONCLUSIONS: It was observed that [beta]B2- and [gamma]C crystallin showed more similar spectroscopic and unfolding properties with each other than each of them showed with [alpha]A-crystallin. PMID- 11483895 TI - Early postoperative feeding after major gynecologic surgery: evidence-based scientific medicine. AB - Postoperative gastrointestinal care after major gynecologic surgery has evolved considerably over the last decade. According to evidence-based scientific medicine, the following conclusions can be drawn: (1) Postoperative colonic stasis occurs after major abdominal surgery and persists for approximately 3 days (classes I and IIA). (2) Elective postoperative nasogastric decompression after major abdominal surgery is unnecessary (class I). (3) Early feeding after major gynecologic surgery results in emesis but does not increase the incidence of aspiration pneumonia, dehiscence, or intestinal leaks and decreases hospital stay (class I). (4) Slow advancement of postoperative diet after major gynecologic surgery is probably unnecessary (class III). (5) After major abdominal gynecologic surgery, there appear to be minimal medical benefits (decreased infection rate) of early postoperative feeding (class III). (6) After radical hysterectomy, postoperative bowel stimulation decreases length of hospital stay (class IIA). PMID- 11483896 TI - Vitamins C and E: missing links in preventing preterm premature rupture of membranes? AB - We propose that generation of reactive oxygen species may be a potentially reversible pathophysiologic pathway leading to preterm premature rupture of the membranes. Reactive oxygen species generated by the body's response to diverse insults such as infection, cigarette smoking, bleeding, or cocaine use can activate collagenolytic enzymes and impair fetal membrane integrity. Vitamin E, a lipid-soluble antioxidant, inhibits membrane-damaging effects of reactive oxygen species-induced lipid peroxidation. Vitamin C, a water-soluble antioxidant in plasma, stimulates and protects collagen synthesis while recycling vitamin E. Prior evidence shows that (1) damage by reactive oxygen species can impair fetal membrane integrity, (2) reduced midgestation levels of vitamin C are associated with preterm premature rupture of membranes, and (3) these vitamins can be safely and effectively absorbed and delivered to gestational tissues. Current prenatal vitamin preparations contain vitamins C and E in concentrations that are less than 1/3 and 1/10, respectively; these levels have been suggested for effective antioxidant protection. We hypothesize that increased dietary consumption or supplementation of vitamins C and E during pregnancy may reduce physiologically the risks of that portion of preterm premature rupture of membranes that is mediated by excessive or undamped peroxidation of fetal membranes. This hypothesis, if confirmed, should stimulate initiation of therapeutic trials to test the efficacy of enhanced supplementation with vitamins C and E during pregnancy to prevent preterm premature rupture of membranes. PMID- 11483897 TI - Two- and 3-dimensional MRI comparison of levator ani structure, volume, and integrity in women with stress incontinence and prolapse. AB - OBJECTIVE: The aim of this study was to identify imaging markers for genuine stress incontinence and pelvic organ prolapse by using magnetic resonance imaging and reconstructed 3-dimensional models. STUDY DESIGN: Thirty women were studied, 10 with prolapse, 10 with genuine stress incontinence, and 10 asymptomatic volunteers. Axial and sagittal T1 and T2 weighted pelvic magnetic resonance scans were obtained with the patient in the supine position. Source images were measured to determine levator hiatus height, bladder neck to pubococcygeal line, levator plate angle, and perineal descent at rest and maximum Valsalva. Manual segmentation and surface modeling was applied to build 3-dimensional models of the organs. The 3-dimensional models were measured to determine levator muscle volume, shape and hiatus width, distance between symphysis and levator sling muscle, posterior urethrovesical angle, bladder neck descent, and levator plate angle. RESULTS: The 3 groups of subjects were comparable in age, parity, and body mass index. In the control, genuine stress incontinence, and prolapse groups, the menopausal rate was 40%, 60%, and 55% (P =.7). In the same order, significant mean 2-dimensional measures were: resting bladder neck descent of 24, 17, and 3 mm (P <.005), straining levator plate angle of -4.3, -11.5, and -31 degrees (P =.01), straining levator hiatus height of 48.5, 51.1, and 65.3 mm (P <.005), and straining perineal descent of 17.2, 22.5, 27.2 mm (P =.02). Similarly ordered mean 3-dimensional parameters showed levator volumes of 32.2, 23.3, and 18.4 cm(3) (P <.005); hiatus widths of 25.7, 34.7, and 40.3 mm (P <.005); left levator sling muscle gaps of 15.6, 20.3, and 23.8 mm (P =.03), right levator sling muscle gaps of 15.6, 22.5, and 30.8 mm, (P = 0.003), and levator shape (90%, 40%, and 20% dome shaped; P <.005). CONCLUSION: Both 2-dimensional magnetic resonance images and 3-dimensional models yield findings that differ among asymptomatic subjects compared with those with genuine stress incontinence and prolapse. Our 3 dimensional data demonstrate a statistically significant continuum in levator volume, shape, and integrity across groups of asymptomatic, genuine stress incontinence, and prolapse subjects. PMID- 11483898 TI - Responsiveness of quality of life measurements to change after reconstructive pelvic surgery. AB - OBJECTIVE: We sought to determine whether Incontinence Impact Questionnaire (IIQ 7) and Urogenital Distress Inventory (UDI-6) scores are sensitive to changes in clinical status after surgery for genuine stress incontinence (GSI) or pelvic organ prolapse (POP). STUDY DESIGN: Patients were included in this study if they had completed the IIQ-7 and UDI-6 scales and had undergone urodynamic testing before and 3 months after surgery for GSI or POP. Kruskal-Wallis tests and logistic regression were used to compare IIQ-7 and UDI-6 scores according to the outcome of surgery. RESULTS: Among 55 patients satisfying inclusion criteria for the study, 34 (62%) underwent Burch colposuspension, and 21 (38%) underwent suburethral sling procedures for GSI; 44 (80%) patients were subjectively continent after surgery. Thirty-four (62%) patients underwent surgical treatment of their POP; 31 (90%) were subjectively cured of their POP symptoms. Mean IIQ-7 and UDI-6 scores were lower in patients who were subjectively continent, and UDI 6 scores were lower in patients who were objectively cured of GSI. CONCLUSION: UDI-6 and IIQ-7 scores change after surgery for GSI and POP, with patients who are subjectively continent having lower postoperative scores on both scales. PMID- 11483899 TI - The Incontinence Impact Questionnaire and the Urogenital Distress Inventory: a revisit of their validity in women without a urodynamic diagnosis. AB - OBJECTIVES: We sought to assess validity of the Incontinence Impact Questionnaire (IIQ) and the Urogenital Distress Inventory (UDI) (long and short forms) in incontinent women previously urodynamically undiagnosed. STUDY DESIGN: Post hoc analysis of responses to the IIQ and UDI questionnaires were obtained from a trial on a urethral device in community-dwelling incontinent women. Internal consistency and validity were evaluated against the 1-hour pad test. RESULTS: Internal consistency (Cronbach's alpha) for the long forms was high for the IIQ and moderately high for the UDI, good for the IIQ-short, but poor for the UDI short. Correlations with 1h pad test were low and non-significant for both versions. Correlation of the short with the long forms was high. CONCLUSION: In the community-dwelling population, without a urodynamic diagnosis, neither long nor short versions of the questionnaires correlate with the severity of the urinary incontinence as shown by the pad test. The validity of the current questionnaires in women without urodynamic diagnosis is questionable. PMID- 11483900 TI - Mersilene mesh sling: short- and long-term clinical and urodynamic outcomes. AB - OBJECTIVE: We sought to determine the long-term efficacy, safety, and urodynamic effects of the Mersilene mesh suburethral sling in treating complicated forms of genuine stress incontinence. STUDY DESIGN: Two hundred women diagnosed with genuine stress incontinence, complicated by recurrence, intrinsic sphincter deficiency, or chronically increased intraabdominal pressure underwent a suburethral mesh sling procedure (Mersilene; Ethicon Inc, Somerville, NJ). They were monitored with yearly clinical examinations plus short- and long-term postoperative urodynamic evaluations; statistical analysis was carried out by use of the Friedman 2-way analysis by rank, Fischer-Freeman-Halton exact testing, analysis of variance for repeated measures, Wilcoxon, exact Mann-Whitney U test, and Bonferroni paired t test. Of 176 patients who were 5 months or more postop, 127 (72%) had preoperative and short-term postoperative urodynamic evaluations (range 5 to 23 months, mean 12.6 months). Fifty-two of 117 women who were more than 19 months postop (44%) completed preoperative and long-term postoperative urodynamic evaluations at a mean of 63 months (range 20 to 107). One hundred thirty-six of 176 patients (77%) who were more than 4 months postop had a short- and/or long- term postoperative urodynamic evaluation (range 5 to 107 months, mean 30 months). RESULTS: Objective cure rate by stress test was 93% (126 of 136 patients) at a mean of 30 months follow-up. The long-term objective cure rate was 94% (49 of 52). Subjectively, the short- and long-term cure rates were 95.3% and 90.4%, respectively. The cotton swab angle deflection decreased by a mean of 54 degrees at 1 year and 50 degrees at 5 years. Of the 10 failures, the mean preoperative cotton swab straining angle was 19.6 degrees, with 6 being < 30 degrees. Nineteen patients had a negative preoperative cotton swab angle test result (mean straining angle 15 degrees before operation, -6 degrees after operation) and a long-term cure rate of 67%. The objective cure rate in patients with positive cotton swab angle results monitored long term (mean 62 months) was 100% (41 of 41). The postvoid residual increased by a mean of 25 mL short term and 10 mL long term. Thirty-eight patients (19%) had a total of 43 complications. Seven patients (3.5%) had long-term retention. De novo detrusor instability occurred in 12 patients (8.8%), although it was cured in 6 (4.4%). Eight patients (4%) had vaginal or inguinal sling erosion and were healed after revision. Delayed healing at the vaginal sling site responded completely to estrogen cream in two (1%) patients. Five women had treatable vaginal stenosis, 5 had a local inguinal collection/infection unrelated to the mesh, and 3 required a 2-unit transfusion of packed red blood cells. One patient each had an entrapped nerve released, a cystotomy repaired, or experienced thigh numbness or groin pain. CONCLUSIONS: The suburethral Mersilene mesh sling has a very high long-term objective and subjective cure rate in the treatment of complicated forms of genuine stress incontinence. Frequent complications do occur but are remediable. The 33% failure rate among patients with a preoperative negative cotton swab angle test result and the very low cotton swab straining angle among the 7% who had sling failures further confirms the widely held belief that sling urethropexy in the absence of hypermobility lacks efficacy. PMID- 11483901 TI - Attachment of the rectovaginal septum to the pelvic sidewall. AB - OBJECTIVE: To describe the lateral attachment of the rectovaginal fascia to the pelvic sidewall. STUDY DESIGN: A descriptive study was performed with use of 10 embalmed female cadaveric pelves, each sectioned in the midsagittal plane. The lateral attachments of the pubocervical fascia and the rectovaginal fascia to the pelvic sidewall were examined. RESULTS: The rectovaginal fascia attaches to the pelvic sidewall along a well-defined line. It extends from the perineal body toward the arcus tendineus fasciae pelvis with which it converges approximately midway between the pubis and the ischial spine to form a y configuration. This point of convergence occurs an average of 4.8 cm from the ischial spine, 3.75 cm from the pubic symphysis, and 4.15 cm from the posterior fourchette. CONCLUSION: The rectovaginal fascia supports the posterior compartment analogous to the pubocervical fascia in the anterior compartment. Moreover, landmarks are identified that will aid suture placement during repair of posterior compartment defects. PMID- 11483902 TI - A structured system to evaluate urethral support anatomy in magnetic resonance images. AB - OBJECTIVE: To develop a systematic method for analyzing the normal magnetic resonance imaging location and appearance of structural features involved in urethral support. STUDY DESIGN: Multiplanar proton density magnetic resonance images of 50 nulliparous women were made at 0.5-cm intervals. The arcuate pubic ligament was the chosen reference point in all views. Structural features were located by identification of the magnetic resonance images in which they appeared. The presence or absence of urethral support structures in each scan level relative to the arcuate pubic ligament was evaluated and recorded as a graphic display. Support structures examined were the arcus tendineus fasciae pelvis, the perineal membrane, the pubococcygeal levator ani muscle and its vaginal and bony attachments, and the pubovesical muscle. Structural definitions were developed on the basis of established periurethral anatomy. Two examiners independently assessed 10 scans for interobserver concordance. This system was used in nulliparous women to quantify the location of magnetic resonance visible structures. RESULTS: Because all levels were 0.5 cm apart, slice identification represented the distance above the arcuate pubic ligament (eg, 2 images above the arcuate pubic ligament or 1.0 cm). Interobserver concordance was 88% in identification of structure location. In the axial plane, specific structural features and relationships were localized. The frequency at which a specific structure was observed at a specific image level in all women was depicted as a gray density within the graphic display. These frequencies indicated where we would expect each structure to be located in healthy women. Relationships between structures and their attachments became apparent. Consideration of expected structural locations proven by nulliparous scans will enable us to quantitatively define abnormal structures in parous women. CONCLUSIONS: This systematic magnetic resonance evaluation allows, for the first time, quantification of the normal anatomic location of urethral support structures. It can be used to help identify the difference between structural abnormalities and normal variation in parous women. PMID- 11483903 TI - Role of urethral electromyography in predicting outcome of Burch retropubic urethropexy. AB - OBJECTIVE: To establish preoperative urethral electromyographic parameters that predict which women are unlikely to be cured of genuine stress incontinence by Burch retropubic urethropexy procedures. STUDY DESIGN: Eighty-nine women who underwent preoperative urodynamic testing with urethral electromyography and retropubic urethropexy for genuine stress incontinence were prospectively studied. Raw electromyographic signals were processed by an electromyographic instrument equipped with automated motor unit analysis software programs. Quantitative electromyographic software was used to analyze the electrical activity of the urethral sphincter with use of mean rectified voltage (MRV)-mean amplitude calculated over the entire tracing after the waveform is rectified-with women at rest and during voluntary urethral squeezing, repetitive coughing, and bladder filling. Objective outcomes were determined 3 months after the operation with single-channel cystometrograms performed while subjects were standing. Nonparametric statistical analyses included the chi(2) test of association for nominal data and the Mann-Whitney test for comparison of population medians. RESULTS: All women had urethral hypermobility and met our standard clinical criteria for retropubic urethropexy. Fifteen women had incomplete follow-up data and were excluded from final analysis. Fifty-nine of 74 women (80%) were objectively cured, and 15 women had persistent genuine stress incontinence. Women who were cured did not differ from those who were not cured in age, parity, menopausal status, maximum urethral closure pressure, Valsalva leak point pressure, maximum cystometric capacity, detrusor instability, or prolapse stage. Women with persistent genuine stress incontinence were more likely to have had previous pelvic operations (P =.01). There were no differences in any electromyographic parameters at rest, with urethral squeezing, or during bladder filling between the groups. Women who were objectively cured had larger MRV values with repetitive coughing (P =.05) and larger increases from resting MRV values (DeltaMRV) with repetitive coughing (P =.04). Twenty-seven of 30 women with MRV values > or =25 microV with repetitive coughing were cured (positive predictive value [PPV] = 90%; negative predictive value [NPV] = 32%), and 22 of 24 women with more than a 10-microV increase in resting MRV values with repetitive coughing (DeltaMRV > 10 microV) were cured (PPV = 92%; NPV = 29%). If women had both an MRV value > or =25 microV and a DeltaMRV value >10 microV, the PPV was 100%; however, the NPV remained at 30%. CONCLUSION: Women who were cured of genuine stress incontinence with Burch retropubic urethropexy procedures had better motor unit action potential activation with repetitive coughing than women with persistent genuine stress incontinence. Urethral electromyography may be used to assess the neuromuscular integrity of the striated urethral sphincter and to help predict which women will have successful retropubic urethropexy procedures. PMID- 11483904 TI - Tolterodine versus oxybutynin in the treatment of urge urinary incontinence: a meta-analysis. AB - OBJECTIVE: To compare tolterodine with oxybutynin in treatment of urge incontinence. STUDY DESIGN: A systematic review, following Cochrane methods, was performed to retrieve results of randomized trials that compared tolterodine with oxybutynin in adults with urge incontinence. Composite point estimates of efficacy (episodes of incontinence per 24-hour period, frequency, and voided volume) and safety (dry mouth, withdrawal, and dose modification) were calculated. RESULTS: Four studies were included. Both drugs similarly decreased the number of micturitions in a 24-hour period. Oxybutynin was marginally superior to tolterodine in decreasing the number of incontinent episodes in a 24 hour period (weighted mean difference, 0.41; 95% confidence interval [CI], 0.04 to 0.77) and increasing the mean voided volume per micturition (8.24 mL; 95% CI, 14.19 to 3.38). Fewer patients had dry mouth (relative risk, 0.54; 95% CI, 0.48 to 0.61) and withdrew from the study because of side effects (relative risk, 0.63; 95% CI, 0.46 to 0.88) with tolterodine. CONCLUSIONS: Oxybutynin and tolterodine share a clinically similar efficacy profile (although oxybutynin is statistically superior), but tolterodine is better tolerated and leads to fewer withdrawals as a result of adverse events. PMID- 11483905 TI - A randomized, double-blind placebo-controlled trial of the effects of the 5 hydroxytriptamine(4) agonist cisapride on the female urinary bladder. AB - OBJECTIVE: Anecdotal reports have proposed the use of the 5-hydroxytriptamine(4) agonist cisapride as a treatment for female voiding dysfunction on the basis of the known prokinetic actions of the medication. The purpose of our study is to assess the effects of this agent on the normal bladder in vivo. STUDY DESIGN: In this randomized, double-blind placebo-controlled trial, patients were randomized to receive either 20 mg cisapride or an identical placebo. They then underwent urodynamic evaluation that included uroflowmetry, multichannel filling cystometry, pressure-flow studies, and a urethral pressure profile. After a washout period of at least 7 days, subjects were then crossed over to the other arm and the tests were repeated. RESULTS: Twenty women without significant urinary incontinence agreed to participate. There was a decrease in the maximum cystometric capacity from 556 mL for placebo to 496 mL for cisapride (P <.001). There was no difference in the detrusor pressure at maximum flow, the maximum detrusor pressure, the flow rate, or the percentage of maximum cystometric capacity voided. CONCLUSIONS: In healthy women, cisapride caused a significant decrease in the maximum cystometric capacity, which could account for the higher reported rates of urinary frequency and urgency with this medicine. There was no evidence that this prokinetic agent improved voiding function. PMID- 11483906 TI - On-screen vector-based ultrasound assessment of vesical neck movement. AB - OBJECTIVE: We sought to develop a vector-based assessment to determine the magnitude and direction of bladder neck movements, as well as to assess whether probe movement relative to the pubis needs to be taken into account. STUDY DESIGN: Ten nulliparous continent, 10 primiparous continent, and 10 primiparous stress-incontinent women were recruited. Perineal ultrasound scanning was performed in standing women while they were resting, performing the Valsalva maneuver, coughing, and performing Kegel exercises. A direct on-screen assessment of bladder neck displacement from rest to the peak of dynamic activity relative to the pubic axis was made. Transducer movement was assessed by measuring the displacement of the pubic bone. RESULTS: The method was feasible because measurements were possible in all 30 subjects. Vesical neck and pubic point movement in millimeters (+/- SD) and the percentage error if pubic point movement is not accounted for are as follow: strain, vesical neck 16.9 +/- 6.1 and pubic point 4.8 +/- 3.9, 28%; cough, vesical neck 10.2 +/- 5.4, pubic point 2.9 +/- 3.4, 33%; Kegel exercise, vesical neck 7.0 +/- 3.6 and pubic point 0.7 +/- 1.4, 37%. Similar discrepancies in angle were found and are presented. Uncorrected direction of vesical neck and pubic point movement in degrees and the percentage error if pubic point movement is not accounted for are as follow: strain, vesical neck 169.4 +/- 18.5 and pubic point 214.0 +/- 56.7, 18%; cough, vesical neck 162.0 +/- 12.8, pubic point 238.4 +/- 27.4, 22%; Kegel exercise, vesical neck 0.9 +/- 12.7 and pubic point -4.8 +/- 20.6, 87%. Test-retest reliability correlations were more than an r value of 0.7 in all measures and 86% of the measurements greater than 0.8. CONCLUSION: The vector-based system provides a simple method for quantifying distance and direction of vesical neck motion, as well as localizing the resting vesical neck position. PMID- 11483907 TI - Assessment and grading of pelvic organ prolapse by use of dynamic magnetic resonance imaging. AB - OBJECTIVE: Our purpose was to assess and compare a new technique of grading pelvic organ prolapse by using dynamic magnetic resonance imaging with the clinical staging proposed by the International Continence Society. STUDY DESIGN: In a cross-sectional study, 20 patients with pelvic organ prolapse underwent dynamic magnetic resonance imaging. Clinical staging (according to the International Continence Society) was compared with staging by magnetic resonance imaging. A new reference line, the midpubic line, was drawn on the magnetic resonance image to correspond to the hymenal ring marker used in the clinical staging. The levator-vaginal angle and the area of the genital hiatus were measured to indicate vaginal supports. Ten nulliparous, symptom-free women were studied as control subjects. RESULTS: The proposed staging by magnetic resonance imaging showed good correlation with the clinical staging (kappa = 0.61). Magnetic resonance imaging improved clinical assessment by its ability to measure the actual pelvic organ descent and to delineate prolapse of the pouch of Douglas accurately. The midpubic line was a useful reference line for grading prolapse on magnetic resonance imaging. The levator-vaginal angle and the area of the genital hiatus are useful in assessing vaginal support at different anatomic levels. CONCLUSIONS: This new method of grading by magnetic resonance imaging uses the same landmarks as the clinical grading, and this uniformity of approach allows an objective assessment of the results of surgical correction of pelvic organ prolapse. PMID- 11483908 TI - Altered immune response in adult women exposed to diethylstilbestrol in utero. AB - OBJECTIVE: Between 1940 and 1970, 1.5 million female fetuses were exposed to diethylstilbestrol in utero. Numerous deleterious effects on reproductive anatomic and physiologic characteristics have been documented in these women. However, the effects of this exposure on nonreproductive systems, which may have lifelong consequences as this cohort of women progresses beyond the childbearing years, have received little attention. On the basis of an earlier preliminary observation of altered immune reponse, we hypothesized that diethylstilbestrol exposed women may show abnormalities in T-cell-mediated immune response. STUDY DESIGN: Thirteen women exposed to diethylstilbestrol in utero were compared with 13 age- and menstrual cycle phase-matched control subjects with respect to the in vitro T-cell response to the mitogens phytohemagglutinin, concanavalin A, and interleukin 2. RESULTS: As compared with controls, tritiated thymidine incorporation by T cells harvested from diethylstilbestrol-exposed women was increased 3-fold over a range of concentrations in response to concanavalin A (P <.001), increased by 50% over a range of concentrations in response to phytohemagglutinin (P <.001), and increased 2-fold in response to the endogenous mitogen interleukin 2 (P <.05). CONCLUSIONS: In vitro evidence suggests that women exposed to diethylstilbestrol have alterations in T-cell-mediated immunity. These changes require further attention with regard to their characterization, their role in the pathogenesis of cancer and autoimmunity, and their presence in normal women exposed to diethylstilbestrol in utero. PMID- 11483909 TI - The risk of stress incontinence 5 years after first delivery. AB - OBJECTIVES: We aimed to evaluate the impact of a first pregnancy and delivery on the prevalence of stress incontinence 5 years afterward. STUDY DESIGN: This longitudinal cohort study included 278 women who were questioned after their first delivery and again 5 years later. RESULTS: The prevalence of stress incontinence 5 years after a first delivery was 30%, and the 5-year incidence was 19%. The risk of stress incontinence 5 years after a first delivery was related to the onset and duration of symptoms after the first pregnancy and delivery in a "dose-response-like" manner. The use of vacuum extraction or episiotomy during the first delivery increased the risk. CONCLUSIONS: First pregnancy and delivery may result in stress incontinence 5 years later. Women with incontinence 3 months after a first delivery have a particularly high risk of long-lasting symptoms. Obstetric risk factors are vacuum extraction and episiotomy. PMID- 11483910 TI - Vascular reactivity in men and women of reproductive age. AB - OBJECTIVE: This study was undertaken to document vascular reactivity in both phases of normal menstrual cycles and anovulatory cycles of women receiving oral contraceptives and to compare vascular reactivity of women in the follicular phase of the normal menstrual cycle with that of men of a similar age group. STUDY DESIGN: Forearm blood flow was measured with plethysmography in 15 women with ovulatory cycles during the follicular and luteal phases of the normal cycle, at equivalent times in 14 women receiving oral contraceptives, and in 15 age-matched men. Changes in forearm blood flow, an index of vascular reactivity, were measured during a randomly ordered brachial artery infusion schedule of norepinephrine, sodium nitroprusside, angiotensin II, and Nomega-monomethyl-L arginine and were analyzed with repeated measures analysis of variance. RESULTS: Forearm blood flow responses were similar in the two phases of the normal cycle. Responses were similar between women with ovulatory cycles and users of oral contraceptives, apart from a significantly greater response to angiotensin II in the oral contraceptive group during the medication phase than during the withdrawal phase (P =.027). Basal forearm blood flow was higher in men than in women (7 +/- 1.6 vs 5 +/- 1.4 mL/100 mL per minute; P =.015), and men demonstrated a significantly greater vasoconstrictor response to norepinephrine (P <.05) than did women. CONCLUSION: Vascular reactivity is similar in the two phases of the normal menstrual cycle, but women who use oral contraceptives have an exaggerated response to angiotensin II during the medication phase. Men demonstrate greater vascular reactivity to norepinephrine than do women, perhaps because of the lack of estrogen-mediated protective effects and increased adrenergic activity. PMID- 11483911 TI - Assessing symptoms before hysterectomy: is the medical record accurate? AB - OBJECTIVE: Our purpose was to evaluate the agreement between the documentation of symptoms leading to hysterectomy and the assessment of those symptoms by the patient. STUDY DESIGN: A retrospective study was performed of 497 women in southern California who had hysterectomies. Sensitivity, specificity, and kappa statistics were calculated for the medical records and were compared with patient interviews for the presence and severity of symptoms. RESULTS: The medical record was 93% sensitive and 61% specific for identifying bleeding and 79% sensitive and 55% specific for identifying pain. Overall agreement between physician records and patient interviews was moderate for bleeding (kappa, 0.55-0.58), fair for pain (kappa, 0.29-0.34), and poor for impairment as a result of bleeding or pain (kappa, 0.0-0.14). CONCLUSIONS: Physician overestimation of symptoms could lead to overuse of hysterectomy, whereas underestimation could result in underuse. Our results suggest that both underestimation and overestimation occur for patients with abnormal bleeding, pain, or both. If physicians accurately assess symptoms but fail to document them, examinations of appropriateness will be faulty unless patients are interviewed. PMID- 11483912 TI - A randomized controlled trial of fentanyl for abortion pain. AB - OBJECTIVE: Our aim was to find out whether intravenous fentanyl was effective in reducing the pain of first-trimester abortion. STUDY DESIGN: This randomized controlled trial included 825 women attending a nonhospital abortion facility. Some women chose standard care. Women who did not choose standard care were randomly assigned to receive either 50 to 100 microg of fentanyl, a placebo, or no intervention. With SAS software and a mixed effects analysis of variance model with covariates, we compared mean pain scores of the fentanyl and placebo groups to detect a difference of at least 1 point on an 11-point pain scale. RESULTS: The mean pain score of the fentanyl group was 1.0 point less than that of the placebo group (95% confidence interval, 3.7-4.3) and 0.9 point less than that of the observational group (95% confidence interval, 4.7-5.1). This pain reduction was statistically significant, but the women who were studied wanted a 2-point reduction from fentanyl. CONCLUSION: Fentanyl, when compared with the placebo, reduced abortion pain by 1.0 point on an 11-point scale. This reduction was of questionable clinical significance and was less than desired by the women included in the study. PMID- 11483913 TI - Microsatellite alterations in exfoliated cervical epithelia deoxyribonucleic acid as a marker for high-grade dysplasia. AB - OBJECTIVE: The purpose of this study was to evaluate the feasibility of the use of deoxyribonucleic acid microsatellite alterations in cervical epithelia in the prediction of high-grade dysplasia and to compare it with a strategy based on human papillomavirus testing. STUDY DESIGN: Our prospective study subjects were from a cohort of 498 women with minimally abnormal Papanicolaou test results including atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesion who had documented repeated Papanicolaou and human papillomavirus tests. Of these, 52 eligible patients having conizations or hysterectomies as their histologic outcomes were subjected to tests of loss of heterozygosity on a panel of 5 microsatellites (D3S1110, THRB, D3S1228, D6S291, D3S1289) within the deoxyribonucleic acid of exfoliated cervical epithelia. These genetic alterations were analyzed through fluorescence polymerase chain reaction by comparison of allele ratios of exfoliated cells with those of normal control tissue. Predictive values for high-grade cervical dysplasia and cancer between this deoxyribonucleic acid marker and human papillomavirus status were compared. RESULTS: With the use of loss of heterozygosity in at least one locus for predicting high-grade cervical neoplastic lesion, the sensitivity, specificity, positive predictive value, and negative predictive value were 96.7%, 59.1%, 76.3%, and 92.9%, which were superior to those of the human papillomavirus test (80%, 59.1%, 72.7%, and 92.9%). As a triage for atypical squamous cells of undetermined significance, its sensitivity and negative predictive value were up to 100%. CONCLUSION: The promising results on determining microsatellite alteration in dysplastic lesions might imply that it is possible to detect the earliest changes by potential molecular markers with exfoliated cervical epithelial cells. PMID- 11483914 TI - Race as a predictor of urinary incontinence and pelvic organ prolapse. AB - OBJECTIVE: The purpose of this study was to assess the effect of race on the results of a standardized evaluation of urinary incontinence and prolapse in African Americans and Caucasians. STUDY DESIGN: This was an analysis of 183 African Americans and 132 Caucasians referred consecutively for symptoms associated with urinary incontinence, pelvic organ prolapse, or both. RESULTS: Genuine stress incontinence, detrusor instability, mixed incontinence, or other condition was diagnosed in 22%, 30%, 16%, and 32% of the African-American subjects, respectively, compared with 46%, 13%, 11%, and 31% of the Caucasian subjects (P =.001). There was no significant racial difference in the presence or severity of pelvic organ prolapse. With the use of stepwise logistic regression to compare risk factors for incontinence, Caucasian race was the most significant predictor of genuine stress incontinence (odds ratio 2.21; 95% confidence interval 1.31-3.73), and African-American race was the only significant predictor of detrusor instability (odds ratio 2.6; 95% confidence interval 1.45-4.80). CONCLUSION: Risk factors for and conditions of urinary incontinence differed significantly between African-American and Caucasian women in this population. Racial differences in risk factors and in prevalence of incontinence subtypes may have importance for the diagnosis and prevention of urinary incontinence. PMID- 11483915 TI - Hormonal and barrier contraception and risk of upper genital tract disease in the PID Evaluation and Clinical Health (PEACH) study. AB - OBJECTIVE: Among women diagnosed with pelvic inflammatory disease, we examined the associations between hormonal or barrier methods of contraception and upper genital tract infection or inflammation. METHODS: Participants were 563 patients from a treatment trial for pelvic inflammatory disease. All had pelvic pain; pelvic organ tenderness; and leukorrhea, mucopurulent cervicitis, or untreated cervicitis. Contraceptive use within the prior 4 weeks was compared among women with baseline upper genital tract gonorrhea or chlamydia, women with endometritis without upper genital tract gonorrhea or chlamydia, and women with neither upper genital tract gonorrhea or chlamydia nor endometritis. RESULTS: Inconsistent condom use was significantly and independently associated with a 2 to 3 times elevated risk for upper genital tract infection. Upper genital tract gonorrhea or chlamydia was not significantly associated with use of oral contraceptives, use of medroxyprogesterone, condoms used consistently, nor other barrier methods. CONCLUSION: No hormonal or barrier contraceptive method was related to a reduction in upper genital tract disease among women with clinical pelvic inflammatory diseases. PMID- 11483916 TI - Does epidural analgesia prolong labor and increase risk of cesarean delivery? A natural experiment. AB - OBJECTIVE: More than 50% of pregnant women in the United States are using epidural analgesia for labor pain. However, whether epidural analgesia prolongs labor and increases the risk of cesarean delivery remains controversial. STUDY DESIGN: We examined this question in a community-based, tertiary military medical center where the rate of continuous epidural analgesia in labor increased from 1% to 84% in a 1-year period while other conditions remained unchanged-a natural experiment. We systematically selected 507 and 581 singleton, nulliparous, term pregnancies with spontaneous onset of labor and vertex presentation from the respective times before and after the times that epidural analgesia was available on request during labor. We compared duration of labor, rate of cesarean delivery, instrumental delivery, and oxytocin use between these two groups. RESULTS: Despite a rapid and dramatic increase in epidural analgesia during labor (from 1% to 84% in 1 year), rates of cesarean delivery overall and for dystocia remained the same (for overall cesarean delivery: adjusted relative risk, 0.8; 95% confidence interval, 0.6-1.2; for dystocia: adjusted relative risk, 1.0; 95% confidence interval, 0.7-1.6). Overall instrumental delivery did not increase (adjusted relative risk, 1.0; 95% confidence interval, 0.8-1.4), nor did the duration of the first stage and the active phase of labor (multivariate analysis; P >.1). However, the second stage of labor was significantly longer by about 25 minutes (P <.001). CONCLUSION: Epidural analgesia during labor does not increase the risk of cesarean delivery, nor does it necessarily increase oxytocin use or instrumental delivery caused by dystocia. The duration of the active phase of labor appears unchanged, but the second stage of labor is likely prolonged. (Am J Obstet Gynecol 2001;185:128-34). PMID- 11483917 TI - Utility of Doppler velocimetry in predicting outcome in twin reversed-arterial perfusion sequence. AB - OBJECTIVE: The aim of this study was to describe Doppler velocimetric findings in pregnancies complicated by the twin reversed-arterial perfusion sequence and to determine the association of these findings with pregnancy outcome. STUDY DESIGN: Six twin pregnancies complicated by twin reversed-arterial perfusion sequence had ultrasonographic and Doppler studies performed between 1990 and 1997. Pulsatile vessels in the umbilical cords of the pump and acardiac twins were insonated, and reversal of flow was confirmed in all cases. Resistive index values were calculated, and the difference in resistive index between the pump and acardiac twin in each pair was evaluated as a marker of pregnancy outcome. RESULTS: Five of 6 pump twins survived the immediate neonatal period. Although 5 of the acardiac twins had abnormally elevated Doppler index values, no ratio of systolic to diastolic velocity or resistive index value of the acardiac twin alone was associated with either a good or poor prognosis for the pump twin. Among the 3 pump twins with good outcomes, all had a resistive index difference >0.20. Among the 3 pump twins with poor outcomes, all had small resistive index differences (<0.05). CONCLUSION: We found larger differences in resistive index to be associated with improved outcome of the pump twin in pregnancies complicated by twin reversed-arterial perfusion sequence. Smaller resistive index differences were associated with poor outcome, including cardiac failure and central nervous system hypoperfusion. PMID- 11483918 TI - A population-based case-control teratologic study of ampicillin treatment during pregnancy. AB - OBJECTIVE: This was a study of the association between ampicillin treatment during pregnancy and prevalence of different congenital abnormalities. STUDY DESIGN: The paired analysis of case patients with congenital abnormalities and matched population control subjects was performed in the population-based Hungarian Case-Control Surveillance of Congenital Abnormalities between 1980 and 1996. Of 38,151 pregnant women who had babies without any defects (population control group), 2632 (6.9%) had been treated with ampicillin. Of 22,865 pregnant women who had offspring with congenital abnormalities (case patients), 1643 (7.2%) had been treated with ampicillin (crude odds ratio, 1.0; 95% confidence interval, 0.7-1.2). Of 812 mothers who were delivered of babies affected by Down syndrome (patient control subjects), 61 (7.5%) had ampicillin treatment, and these were also compared with the case group. RESULTS: The prevalence of ampicillin use during the second and third months of gestation, which is the critical period for most major congenital abnormalities, showed significant difference in the case-control pair analysis only for cleft palate (odds ratio, 4.2; 95% confidence interval, 1.4-16.3). This possible association was confirmed by the analysis of medically recorded ampicillin use and by the comparison of ampicillin treatment between the group with cleft palate and the patient control subjects. CONCLUSION: Treatment with ampicillin during pregnancy may pose little if any teratogenic risk in human beings. Only a higher prevalence of cleft palate was found after the ampicillin treatment during the second and third months of gestation. The lack of an experimental animal model and the lack of consistency with previous epidemiologic studies may indicate that even this apparent risk is not real and instead is a chance association; further investigation is therefore necessary. PMID- 11483919 TI - Heparin-induced thrombocytopenia is rare in pregnancy. AB - OBJECTIVE: The indications for heparin use during pregnancy are expanding; however, heparin is associated with serious adverse effects including heparin induced thrombocytopenia. Low-molecular-weight heparin is expensive but is associated with less frequent occurrences of heparin-induced thrombocytopenia in the nonpregnant population. However, the incidence of heparin-induced thrombocytopenia during pregnancy is unknown. The purpose of this study was to compare the incidence of heparin-induced thrombocytopenia in pregnant and nonpregnant women. STUDY DESIGN: This is a retrospective cohort comparison. Pregnant and nonpregnant women were identified by means of diagnosis related group and Current Procedural Terminology code searches at three medical centers in Utah; the incidence of heparin-induced thrombocytopenia in the two groups was compared. RESULTS: There were 10 (4%) cases of thrombocytopenia among 244 heparin treated pregnant patients and 26 (11%) cases among the 244 nonpregnant controls. There were no cases of heparin-induced thrombocytopenia in the pregnant group, but there were 10 (4%) cases in the control group (P =.0014). CONCLUSION: Heparin induced thrombocytopenia is extremely rare in pregnant women. PMID- 11483920 TI - Maternal and fetal inherited thrombophilias are not related to the development of severe preeclampsia. AB - OBJECTIVE: Thrombotic vascular disease may predispose patients to the development of preeclampsia. The purpose of this study was to determine whether maternal or fetal genotype frequencies of the inherited thrombophilic gene mutations (factor V Leiden, methylenetetrahydrofolate, and prothrombin) are altered in severe preeclampsia. STUDY DESIGN: We performed a prospective cross-sectional study to compare the maternal and fetal genotype frequencies of factor V Leiden, methylenetetrahydrofolate, and prothrombin. One hundred ten patients with severe preeclampsia were matched for gestational age to 97 normotensive pregnancies. Umbilical cord blood was obtained from 92 control patients and 75 patients with preeclampsia. Deoxyribonucleic acid was extracted from leukocytes and polymerase chain reaction was performed. Polymerase chain reaction products were digested with the appropriate restriction enzyme and fractionated by gel electrophoresis. Genotype frequencies were calculated. Statistical significance was determined by the chi(2) test. RESULTS: There were no significant differences between patients with severe preeclampsia and control patients regarding frequency of maternal factor V Leiden G/506/A mutation (4.4% vs 4.3%; P =.96), methylenetetrahydrofolate CC/667/TT mutation (9.6% vs 6.3%; P =.54), or prothrombin G/20210/A mutation (0% vs 1.1%; P =.92). In addition, no statistical difference could be found between fetal thrombophilias and the development of preeclampsia. Findings were similar in both white (n = 47) and African American (n = 63) preeclamptic subsets. Moreover, there was no association between any of the maternal or fetal genetic polymorphisms and the incidence of hemolysis, elevated liver enzymes, and low platelet count syndrome (n = 21); eclampsia (n = 12); or intrauterine growth restriction (n = 9). CONCLUSION: Inherited thrombophilias are not associated with severe preeclampsia. PMID- 11483921 TI - Contractility of late pregnant rat myometrium is refractory to activation of soluble but not particulate guanylate cyclase. AB - OBJECTIVE: Our purpose was to compare the effects of agents activating particulate or soluble guanylate cyclases on the spontaneous contractile activity of the isolated pregnant rat uterus. STUDY DESIGN: Uterine rings from midpregnant (14-day) and late pregnant (21-day) rats were suspended in organ chambers to record spontaneous contractile activity. Concentration-response curves were obtained for the following natriuretic peptides: atrial, brain, and C-type; concentration-response curves were also obtained for diethylamine nitric oxide, 3 morpholino-sydnominine, and authentic nitric oxide. RESULTS: All 3 natriuretic peptides inhibited spontaneous uterine contractions equally at midgestation and late gestation. The inhibitory effects of the nitric oxide donors diethylamine nitric oxide, 3-morpholino-sydnominine, and authentic nitric oxide were attenuated in uterine tissues from animals in late stages of pregnancy. CONCLUSION: Agents activating either soluble or particulate guanylate cyclase inhibit contractions of uterine rings from midgestation rats, whereas the effects of soluble guanylate cyclase are attenuated at late pregnancy. Thus spontaneous uterine contractions are under the control of both soluble and particulate guanylate cyclases; the former is dependent on gestational age but the latter is not. PMID- 11483922 TI - Midtrimester vaginal Mycoplasma genitalium in women with subsequent spontaneous preterm birth. AB - OBJECTIVE: We sought to determine the midtrimester prevalence of Mycoplasma genitalium in women who had subsequent spontaneous preterm birth. STUDY DESIGN: In a prospective study of lower genital tract infections, we identified 127 women who subsequently had spontaneous preterm birth. Vaginal samples were obtained between 21 and 25 weeks' gestation for pH, for bacterial vaginosis Gram stain, and cultures that yielded Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis. M genitalium was identified by using validated polymerase chain reaction (PCR) primers, and the results were compared to pregnancy outcomes. RESULTS: Of 124 women with spontaneous preterm births, only five (3.9%) had PCR assays positive forM genitalium. The mean +/- SD delivery gestational age was similar for women with a positive PCR (34.6 +/- 2.2 weeks) and a negative PCR (34.0 +/- 2.7 weeks) (P =.62). None of the women with positive PCR results tested positive for any other sexually transmitted disease, whereas 36 (30%) women with negative PCR results tested positive. CONCLUSIONS: The occurrence of M genitalium in the vagina at midtrimester is infrequent in women with subsequent spontaneous preterm birth. PMID- 11483923 TI - Lipid hydroperoxides and free radical scavenging enzyme activities in preeclampsia and HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome: no evidence for circulating primary products of lipid peroxidation. AB - OBJECTIVE: Preeclampsia is reportedly accompanied by increased plasma lipid peroxidation. However, to date the assays that have been applied to measure lipid peroxides have been relatively nonspecific. A highly specific high-performance lipid chromatographic-chemiluminescence technique was used here to measure lipid hydroperoxides in plasma and red blood cells of patients with preeclampsia and HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome. STUDY DESIGN: Lipid peroxidation products, both lipid hydroperoxides and thiobarbituric acid reactive substances (TBARS), and free radical scavenging enzyme activities were measured in either plasma or erythrocytes of patients with normal pregnancies (n = 36), preeclampsia (n = 28), or HELLP syndrome (n = 10). RESULTS: Whereas lipid hydroperoxides were undetectable (<0.02 micromol/L), plasma TBARS and glutathione peroxidase activity in plasma and erythrocytes were significantly increased in patients with HELLP syndrome. There were no differences in glutathione transferase and reductase activities or superoxide dismutase activity between the groups. CONCLUSIONS: We were unable to corroborate results of studies that suggested that increased TBARS are representative of circulating plasma lipid hydroperoxides in preeclampsia. However, elevated glutathione peroxidase activity and TBARS are in line with increased oxidative stress in preeclampsia. PMID- 11483924 TI - Chorioamnionitis and inflammation of the fetal lung. AB - OBJECTIVE: Fetal intrauterine exposure to proinflammatory cytokines present in amniotic fluid has been associated with an increased risk of chronic lung disease. However, the impact of histologically confirmed chorioamnionitis on the fetal lung has not yet been elucidated. We therefore investigated cellular immune response, cell proliferation, and messenger ribonucleic acid cytokine expression in fetal pulmonary tissue in the presence or absence of chorioamnionitis. STUDY DESIGN: Serial tissue sections were obtained from 27 fetuses at the time of autopsy. Three mothers had received antibiotics for treatment of clinical chorioamnionitis before abortion. Immunohistochemical staining of lung tissue comprised lineage-specific markers (CD68(+), CD3(+), neutrophil elastase). Positively stained cells were evaluated with a graticule, and cells per 5 mm(2) were counted. We undertook in situ hybridization to assess the expression of interleukin 8 messenger ribonucleic acid in the fetal lung. RESULTS: Seven of 27 fetuses had been exposed to chorioamnionitis. Fetal lungs showed a marked increase in the presence of histologically confirmed chorioamnionitis in densities of CD68(+) macrophages (68 vs 9.5 cells/5 mm(2), median group vs control group; P =.02) and lymphocytes (7 vs 2.5 cells/5 mm(2), median chorioamnionitis vs control group; P =.05) and a similar but lesser increase in neutrophil density (16 vs 4 cells/5 mm(2); difference not significant). Interleukin 8 messenger ribonucleic acid was expressed in 4 of 6 tissue specimens investigated in the chorioamnionitis group. Exposure to chorioamnionitis resulted in interleukin 8 messenger ribonucleic acid expression 7-fold higher than in the nonchorioamnionitis group; however, this difference did not achieve statistical significance. CONCLUSION: Chorioamnionitis was associated with an intrauterine inflammatory response of the fetal lung characterized by a severe infiltration of macrophages, neutrophils, and lymphocytes and also by an increased expression of interleukin 8 messenger ribonucleic acid. PMID- 11483925 TI - Maternal-fetal transfer of saquinavir studied in the ex vivo placental perfusion model. AB - OBJECTIVE: This study was undertaken to determine the placental transfer of the human immunodeficiency virus protease inhibitor saquinavir. STUDY DESIGN: An ex vivo perfused human placental cotyledon model was used. Ten placental perfusion studies were performed, with concentrations of saquinavir in the maternal compartment ranging from 322 to 2197 ng/mL (within reference therapeutic ranges). Drug concentrations were determined by high-performance liquid chromatography. RESULTS: The mean (+/- SD) fetal transfer rate of saquinavir was 1.8% +/- 1.6%, and the mean (+/- SD) clearance index was 0.05 +/- 0.05. A mean (+/- SD) of 1.6% +/- 3.1% of the perfused saquinavir was retained by the cotyledon. The small amount of saquinavir that crossed the placenta corresponded to the fraction not bound to human serum albumin. CONCLUSION: The low rate of placental transfer of saquinavir suggests that use of this antiretroviral drug by pregnant women may not lead to significant exposure of the fetus. PMID- 11483926 TI - Inhibition of cyclooxygenase but not nitric oxide synthase influences effects on the human omental artery of the thromboxane A2 mimetic U46619 and 17beta estradiol. AB - OBJECTIVE: These experiments were performed to study the influence of endothelial factors on the contractile effect of the thromboxane A(2) analog U46619 and on the relaxant action of 17beta-estradiol on isolated human omental arteries from nonpregnant women, women with normal pregnancies, and women with preeclampsia. STUDY DESIGN: Arterial rings (3 mm) with or without endothelium were suspended in organ chambers filled with Krebs buffer, 37 degrees C, aerated with 5% carbon dioxide in air, pH approximately 7.4, for isometric tension recording. Rings were incubated with indomethacin, N(omega)-nitro-L -arginine, or 17beta-estradiol, alone or in combination. The concentration that produced 50% of maximal effect, the area under the curve, and the maximal effect of U46619, normalized with respect to a reference contraction in response to potassium chloride, were compared. RESULTS: Neither indomethacin nor N(omega)-nitro-L -arginine changed the basal tone of omental artery rings. Neither N(omega)-nitro-L -arginine nor removal of the endothelium affected either the contractile action of U46619 or the relaxant action of 17beta-estradiol. In contrast, indomethacin potentiated the contractile effect of U46619 and abolished the inhibitory effect of 17beta estradiol in rings from all three groups. The effects of U46619 and 17beta estradiol were significantly less in rings from women with normal pregnancy than in those from women with preeclampsia. Tissues from women with preeclampsia demonstrated the highest contractile response to U46619. CONCLUSION: The inhibitory effect of 17beta-estradiol is not due to increased production of endothelial nitric oxide but rather involves inhibitory products of the cyclooxygenase pathway. The effect of indomethacin may result from inhibited production or release of eicosanoids or from sensitization of thromboxane A(2) receptors. PMID- 11483927 TI - The interactive effects of endotoxin with prenatal glucocorticoids on short-term lung function in sheep. AB - OBJECTIVE: Previously we have shown that neonatal lung function in sheep after preterm birth is profoundly enhanced by intra-amniotic injection of endotoxin, with a magnitude at least equal to that induced by maternal betamethasone administration. This study investigated the effects of betamethasone on lung maturation and growth in the presence of inflammation by treating sheep with both maternal intramuscular betamethasone and intra-amniotic endotoxin injections. STUDY DESIGN: Time-mated pregnant ewes at 118 days' gestation were allocated at random to receive maternal intramuscular or intra-amniotic saline solution injection (n = 10), maternal intramuscular betamethasone injection (0.5 mg/kg; n = 7), intra-amniotic endotoxin injection (20 mg Escherichia coli B055;B5; n = 11) by ultrasonographic guidance, or both betamethasone and endotoxin injections (n = 7). The lambs were delivered abdominally at 125 days' gestation (term is 150 days' gestation), and the neonates were ventilated for 40 minutes before postmortem examination. RESULTS: Combined treatment with betamethasone and endotoxin resulted in significantly greater improvements in neonatal lung function than occurred after treatment with either agent alone, and this effect was not accompanied by a further increase in surfactant levels. The reduction in birth weight that is seen after maternal betamethasone treatment was not seen when this treatment was combined with endotoxin. Endotoxin treatment resulted in inflammatory responses in cord blood and alveolar wash, and these responses were not inhibited by betamethasone treatment. There were no pregnancy losses. CONCLUSION: Both intra-amniotic endotoxin injection and maternal intramuscular betamethasone injection promoted fetal lung maturation. When these treatments were combined, there were additive effects on short-term postnatal lung function but not on surfactant levels. Endotoxin negated the growth restriction in sheep caused by maternal betamethasone treatment. These findings provide evidence that the lung maturation induced by glucocorticoids and that induced by endotoxin are mediated by different mechanisms. PMID- 11483928 TI - Microsatellite analysis reveals a high incidence of maternal cell contamination in 46,XX products of conception consisting of villi or a combination of villi and membranous material. AB - OBJECTIVE: With the use of microsatellite analysis, we sought to determine the incidence of maternal cell contamination in 46,XX products of conception consisting of villi or a combination of villi and membranous material. STUDY DESIGN: Deoxyribonucleic acid from cultured fibroblasts of 46,XX products of conception specimens and a corresponding maternal blood sample were obtained from 31 women. Maternal and fetal genotypes for several highly polymorphic microsatellite markers were compared. RESULTS: Maternal cell contamination was present in 26 (89.7%) of the 29 products of conception specimens from which conclusive results were obtained. The contamination appeared to completely obscure the fetal material in 24 of these specimens. CONCLUSIONS: A significant proportion of 46,XX karyotypes from products of conception represents maternal cell contamination. When maternal cells rather than fetal cells are karyotyped, no information is gained regarding the chromosome constitution of the abortus, and genetic counseling regarding recurrence risks for future pregnancies may be inaccurate. Thus laboratories should exercise caution when reporting normal female karyotypes on products of conception and should consider using microsatellite analysis to determine whether 46,XX results are truly representative of the fetal karyotype. PMID- 11483929 TI - Reversed end-diastolic flow in first-trimester umbilical artery: an ominous new sign for fetal outcome. AB - OBJECTIVE: The purpose of this study was to assess the fetal outcome of first trimester pregnancies with reversed end-diastolic flow in the umbilical artery. STUDY DESIGN: Doppler studies in the umbilical artery were carried out in 2970 consecutive pregnancies scanned at 10 to 14 weeks. RESULTS: We observed 11 (0.4%) cases of reversed end-diastolic flow. Of these, an autosomal trisomy was shown in 7 and a congenital heart defect in 2 additional fetuses. Fetal demise was observed in 5 pregnancies, neonatal death was observed in 1, and termination of pregnancy was carried out in 4. Only 1 fetus survived. CONCLUSIONS: Reversed end diastolic flow in first-trimester umbilical artery signals an ominous prognosis even with normal karyotype. PMID- 11483930 TI - Are risk factors the same for small for gestational age versus other preterm births? AB - OBJECTIVES: This article explores whether the impact of social and demographic risk factors for preterm birth differs for small for gestational age preterm births versus other preterm births. STUDY DESIGN: This was a European case control study of the determinants of preterm birth (4700 cases and 6460 controls). Small for gestational age and non-small for gestational age preterm births were compared with a control group of term births; relationships were explored further by stratifying preterm births into subgroups by mode of onset, the presence of hypertension, and gestational age. RESULTS: Of the social and demographic risk factors for preterm birth identified in this sample, high maternal age, smoking, and low and high maternal body mass index have a stronger effect on small for gestational age preterm births. In contrast, obstetric history, maternal education, and marital status have similar effects regardless of birth weight. Hypertension during pregnancy is strongly associated with small for gestational age preterm birth and contributes to an explanation of observed differences. CONCLUSIONS: These results underline the importance of considering fetal growth restriction in the analysis of risk factors for preterm birth. PMID- 11483931 TI - Preoperative planning with magnetic resonance imaging and computerized volume rendering in twin-to-twin transfusion syndrome. AB - Our purpose was to illustrate the feasibility of preoperative planning with magnetic resonance imaging, 3-dimensional reconstruction, and volume-rendering techniques in twin-to-twin transfusion syndrome treated by endoscopic laser ablation of communicating vessels. After ultrasonographic determination of the syndrome and the indications for intervention, 2 patients with an anterior placenta underwent magnetic resonance imaging without the need for maternal or fetal sedation. Raw image data were downloaded into a desktop computer and manipulated with 3-dimensional reconstruction, volume rendering, and surgical navigation software. In both patients a virtual rendering of the fetuses, placenta, and uterus could be manipulated to expose all sides, demonstrate the location of the intertwin membrane, and plan the point of entry and curve of the endoscopic instruments. Preoperative planning and virtual surgical navigation in fetal surgery are now possible, as a result of shorter magnetic resonance imaging acquisition times and volume-rendering software. In this manner an entire virtual endoscopic fetal operation can be performed and fine-tuned before the actual procedure is to take place. PMID- 11483932 TI - Survival, morbidity, and resource use of infants of 25 weeks' gestational age or less. AB - OBJECTIVE: The objective of this study was to examine survival, morbidity, and resource use in a large cohort of extremely preterm infants. STUDY DESIGN: We examined all (n = 754) neonatal intensive care unit admissions born at < or =25 weeks' gestation and inborn deliveries (n = 949) between 22 and 25 weeks' gestation at 17 Canadian neonatal intensive care units. RESULTS: The overall survival rate was 63%, with a range from 14% at 22 weeks' gestation to 76% at 25 weeks' gestation. There was a high incidence of chronic lung disease (33%-51%), > or =grade 3 intraventricular hemorrhage (0%-16%), necrotizing enterocolitis (0% 14%), > or =stage 3 retinopathy of prematurity (27%-55%), nosocomial infection (25%-39%), and multiple gestation (18%-46%). Extremely preterm infants comprise 4% of neonatal intensive care unit admissions but account for 22% of deaths, 20% 60% of major morbidities, 11% of patient days, and 10%-35% of major procedures. Outborn infants had a higher incidence of chronic lung disease, severe retinopathy of prematurity, and intraventricular hemorrhage. CONCLUSION: Extremely preterm infants have a high incidence of mortality and morbidity and consume disproportionate amounts of neonatal intensive care unit resources. PMID- 11483933 TI - Complementary/alternative therapies for premenstrual syndrome: a systematic review of randomized controlled trials. AB - Complementary/alternative therapies are popular with women who have premenstrual syndrome. This systematic review was designed to determine whether use of such therapies is supported by evidence of effectiveness from rigorous clinical trials. Trials were located through searching 7 databases and checking the reference lists of articles. Randomized controlled trials investigating a complementary/alternative therapy in women with premenstrual syndrome published in the peer-reviewed literature were included in the review. Twenty-seven trials were included investigating herbal medicine (7 trials), homeopathy (1), dietary supplements (13), relaxation (1), massage (1), reflexology (1) chiropractic (1), and biofeedback (2). Despite some positive findings, the evidence was not compelling for any of these therapies, with most trials suffering from various methodological limitations. On the basis of current evidence, no complementary/alternative therapy can be recommended as a treatment for premenstrual syndrome. PMID- 11483934 TI - Disuse osteoporosis as evidence of brachial plexus palsy due to intrauterine fetal maladaptation. AB - We report what may be overlooked evidence of the effects of intrauterine maladaptation as a cause of brachial plexus palsy. A case of total brachial plexus palsy in the posterior arm associated with Horner's syndrome and severe demineralization of the bones of the affected arm is analyzed. In this litigated case, a report of marked demineralization of the bones of the affected arm was analyzed by the plaintiff's radiology expert as diagnostic of disuse osteoporosis. The presence of clear-cut evidence of disuse osteoporosis during the early neonatal period is compelling evidence of an intrauterine onset of brachial plexus palsy. PMID- 11483935 TI - In utero erythrocyte transfusion for fetal xerocytosis associated with severe anemia and non-immune hydrops fetalis. AB - Hereditary xerocytosis is a rare hemolytic anemia occurring secondary to a defect in cell membrane potassium flux. We report a case of severe fetal anemia and non immune hydrops secondary to hereditary xerocytosis that was managed successfully with in utero erythrocyte and albumin transfusion. PMID- 11483936 TI - Influence of maternal and fetal glucokinase mutations in gestational diabetes. AB - We report 2 insulin-treated pregnancies in a mother with hyperglycemia resulting from a glucokinase gene mutation. The inheritance of a glucokinase mutation in 1 child reduced his intrauterine growth (birth weight less than first percentile) by reducing fetal insulin secretion. We discuss the implications for obstetric management of patients with glucokinase mutations. PMID- 11483937 TI - Successful planned pregnancy following endometrial ablation with the YAG laser. AB - In selected patients who previously have undergone YAG laser ablation of the endometrium and who have demonstrated normal intrauterine architecture, pregnancy as a planned event may be a reasonable and safe option. PMID- 11483938 TI - Biparental contribution to fetal thrombophilia in discordant twin intrauterine growth restriction. AB - Poor pregnancy outcomes have been reported to be associated with maternal thrombophilia. We present a case where 1 dizygotic twin inherited thrombophilic genes from both the father and mother, resulting in placental fetal thrombotic vasculopathy and intrauterine growth restriction, whereas its co-twin inherited only 1 such gene from its mother and was unaffected. PMID- 11483939 TI - Changes in biochemical parameters of bone turnover and bone mineral density in post-pregnancy osteoporosis. AB - A lactating 32-year-old woman in whom severe back pain developed 5 months after delivery is described. Cross-linked carboxyterminal telopeptide of type I collagen and pyridinoline, which are parameters of bone resorption, was markedly elevated with low bone mineral density. A high level of bone resorption was associated with post-pregnancy osteoporosis. PMID- 11483941 TI - Treatment of postoperative urinary retention by elongation of tension-free vaginal tape. AB - Tension-free vaginal tape elongation with a polypropylene intermediate piece was successfully carried out in a patient with urinary retention after an initial tension-free vaginal tape surgical procedure. This resulted in immediate residual free voiding and continence, unchanged at 6 months follow-up. Postoperative urinary retention may thus be treated by elongation instead of dissection of the tension-free vaginal tape. PMID- 11483940 TI - Leiomyoma in the space of Retzius: a report of 2 cases. AB - Leiomyomas most frequently occur in the genitourinary and gastrointestinal system. This report discusses clinical and pathologic findings of 2 leiomyomas in the pubovesical space, a location that has not been described before. Different operative approaches were used for excision. Immunohistochemical examination for the presence of estrogen and progesterone receptors was performed. PMID- 11483942 TI - Posthumous reproduction in a human immunodeficiency virus-discordant couple. AB - A human immunodeficiency virus-discordant couple failed to conceive through in vitro fertilization and intracytoplasmic sperm injection of cryopreserved semen banked by the human immunodeficiency virus-positive partner. The husband subsequently had acquired immunodeficiency syndrome and died. The subsequent transfer of cryopreserved embryos resulted in pregnancy. Both mother and child are human immunodeficiency virus negative. PMID- 11483943 TI - Severe vulvovaginitis associated with intravaginal nystatin therapy. AB - A 64-year-old woman had severe vulvovaginitis develop while she was receiving intravaginal nystatin therapy for Candida glabrata infection. Mucocutaneous adverse effects have rarely been reported with nystatin despite long years of use. This complication should be included in the differential diagnosis of clinical failure of intravaginal nystatin therapy. PMID- 11483944 TI - Twin birth weight discordance and the risk of preterm birth. PMID- 11483946 TI - Dubious validity of novel maternal-fetal outcome model. PMID- 11483947 TI - Disappointment with maternal-fetal outcome model. PMID- 11483949 TI - Maternal age and uterine dysfunction. PMID- 11483951 TI - Appendicitis during pregnancy. PMID- 11483953 TI - SV2 modulates the size of the readily releasable pool of secretory vesicles. AB - The exocytosis of neurotransmitters is regulated by calcium and is plastic - features that suggest specialized regulation of the basic membrane trafficking process. Here we show that Synaptic Vesicle Protein 2 (SV2), a protein specific to neurons and endocrine cells, is required to maintain a pool of vesicles available for calcium-stimulated exocytosis. Direct measures of exocytosis in adrenal chromaffin cells showed that the calcium-induced exocytotic burst, which operationally defines the readily releasable pool of vesicles, was significantly reduced in mice lacking SV2A. Burst kinetics were normal in cells from SV2A knockout animals, however, indicating that SV2 functions before the final events of fusion. Analyses of SDS-resistant SNARE (soluble NSF (N-ethylmaleimide sensitive fusion) attachment protein receptor) complexes in brain tissue showed that loss of SV2A was associated with fewer SDS-resistant complexes. Our observations indicate that SV2 may modulate the formation of protein complexes required for fusion and therefore the progression of vesicles to a fusion competent state. PMID- 11483954 TI - ActA and human zyxin harbour Arp2/3-independent actin-polymerization activity. AB - The actin cytoskeleton is a dynamic network that is composed of a variety of F actin structures. To understand how these structures are produced, we tested the capacity of proteins to direct actin polymerization in a bead assay in vitro and in a mitochondrial-targeting assay in cells. We found that human zyxin and the related protein ActA of Listeria monocytogenes can generate new actin structures in a vasodilator-stimulated phosphoprotein-dependent (VASP) manner, but independently of the Arp2/3 complex. These results are consistent with the concept that there are multiple actin-polymerization machines in cells. With these simple tests it is possible to probe the specific function of proteins or identify novel molecules that act upon cellular actin polymerization. PMID- 11483955 TI - TGF-beta-induced apoptosis is mediated by the adapter protein Daxx that facilitates JNK activation. AB - Transforming growth factor-beta (TGF-beta) is a multifunctional growth factor that has a principal role in growth control through both its cytostatic effect on many different epithelial cell types and its ability to induce programmed cell death in a variety of other cell types. Here we have used a screen for proteins that interact physically with the cytoplasmic domain of the type II TGF-beta receptor to isolate the gene encoding Daxx - a protein associated with the Fas receptor that mediates activation of Jun amino-terminal kinase (JNK) and programmed cell death induced by Fas. The carboxy-terminal portion of Daxx functions as a dominant-negative inhibitor of TGF-beta-induced apoptosis in B cell lymphomas, and antisense oligonucleotides to Daxx inhibit TGF-beta-induced apoptosis in mouse hepatocytes. Furthermore, Daxx is involved in mediating JNK activation by TGF-beta. Our findings associate Daxx directly with the TGF-beta apoptotic-signalling pathway, and make a biochemical connection between the receptors for TGF-beta and the apoptotic machinery. PMID- 11483956 TI - Triggering ubiquitination of a CDK inhibitor at origins of DNA replication. AB - To ensure proper timing of the G1-S transition in the cell cycle, the cyclin E Cdk2 complex, which is responsible for the initiation of DNA replication, is restrained by the p21(Cip1)/p27(Kip1)/p57(Kip2) family of CDK (cyclin-dependent kinase) inhibitors in humans and by the related p27(Xic1) protein in Xenopus. Activation of cyclin E-Cdk2 is linked to the ubiquitination of human p27(Kip1) or Xenopus p27(Xic1) by SCF (for Skp1-Cullin-F-box protein) ubiquitin ligases. For human p27(Kip1), ubiquitination requires direct phosphorylation by cyclin E-Cdk2. We show here that Xic1 ubiquitination does not require phosphorylation by cyclin E-Cdk2, but it does require nuclear accumulation of the Xic1-cyclin E-Cdk2 complex and recruitment of this complex to chromatin by the origin-recognition complex together with Cdc6 replication preinitiation factors; it also requires an activation step necessitating cyclin E-Cdk2-kinase and SCF ubiquitin-ligase activity, and additional factors associated with mini-chromosome maintenance proteins, including the inactivation of geminin. Components of the SCF ubiquitin ligase complex, including Skp1 and Cul1, are also recruited to chromatin through cyclin E-Cdk2 and the preinitiation complex. Thus, activation of the cyclin E Cdk2 kinase and ubiquitin-dependent destruction of its inhibitor are spatially constrained to the site of a properly assembled preinitiation complex. PMID- 11483957 TI - mDia mediates Rho-regulated formation and orientation of stable microtubules. AB - Rho-GTPase stabilizes microtubules that are oriented towards the leading edge in serum-starved 3T3 fibroblasts through an unknown mechanism. We used a Rho effector domain screen to identify mDia as a downstream Rho effector involved in microtubule stabilization. Constitutively active mDia or activation of endogenous mDia with the mDia-autoinhibitory domain stimulated the formation of stable microtubules that were capped and oriented towards the wound edge. mDia co localized with stable microtubules when overexpressed and associated with microtubules in vitro. Rho kinase was not necessary for the formation of stable microtubules. Our results show that mDia is sufficient to generate and orient stable microtubules, and indicate that Dia-related formins are part of a conserved pathway that regulates the dynamics of microtubule ends. PMID- 11483959 TI - Valosin-containing protein is a multi-ubiquitin chain-targeting factor required in ubiquitin-proteasome degradation. AB - The ubiquitin-proteasome (Ub-Pr) degradation pathway regulates many cellular activities, but how ubiquitinated substrates are targeted to the proteasome is not understood. We have shown previously that valosin-containing protein (VCP) physically and functionally targets the ubiquitinated nuclear factor kappaB inhibitor, IkappaBalpha to the proteasome for degradation. VCP is an abundant and a highly conserved member of the AAA (ATPases associated with a variety of cellular activities) family. Besides acting as a chaperone in membrane fusions, VCP has been shown to have a role in a number of seemingly unrelated cellular activities. Here we report that loss of VCP function results in an inhibition of Ub-Pr-mediated degradation and an accumulation of ubiquitinated proteins. VCP associates with ubiquitinated proteins through the direct binding of its amino terminal domain to the multi-ubiquitin chains of substrates. Furthermore, its N terminal domain is required in Ub-Pr-mediated degradation. We conclude that VCP is a multi-ubiquitin chain-targeting factor that is required in the degradation of many Ub-Pr pathway substrates, and provide a common mechanism that underlies many of the functions of VCP. PMID- 11483958 TI - The role of Drosophila CID in kinetochore formation, cell-cycle progression and heterochromatin interactions. AB - Centromere function requires the coordination of many processes including kinetochore assembly, sister chromatid cohesion, spindle attachment and chromosome movement. Here we show that CID, the Drosophila homologue of the CENP A centromere-specific H3-like proteins, colocalizes with molecular-genetically defined functional centromeres in minichromosomes. Injection of CID antibodies into early embryos, as well as RNA interference in tissue-culture cells, showed that CID is required for several mitotic processes. Deconvolution fluorescence microscopy showed that CID chromatin is physically separate from proteins involved in sister cohesion (MEI-S332), centric condensation (PROD), kinetochore function (ROD, ZW10 and BUB1) and heterochromatin structure (HP1). CID localization is unaffected by mutations in mei-S332, Su(var)2-5 (HP1), prod or polo. Furthermore, the localization of POLO, CENP-meta, ROD, BUB1 and MEI-S332, but not PROD or HP1, depends on the presence of functional CID. We conclude that the centromere and flanking heterochromatin are physically and functionally separable protein domains that are required for different inheritance functions, and that CID is required for normal kinetochore formation and function, as well as cell-cycle progression. PMID- 11483960 TI - Oncogenes in Ras signalling pathway dictate host-cell permissiveness to herpes simplex virus 1. AB - The importance of herpes simplex viruses (HSV) as human pathogens and the emerging prospect of using mutant derivatives of HSV-1 as potential anti-cancer therapeutics have necessitated a thorough investigation into the molecular basis of host-cell permissiveness to HSV. Here we show that NIH-3T3 cells transformed with the oncogenes v-erbB, activated sos or activated ras become significantly more permissive to HSV-1. Inhibitors of the Ras signalling pathway, such as farnesyl transferase inhibitor 1 and PD98059, effectively suppressed HSV-1 infection of ras-transformed cells. Enhanced permissiveness of the transformed cells was linked to the inhibition of virus-induced activation (phosphorylation) of the double-stranded RNA-activated protein kinase (PKR), thereby allowing viral transcripts to be translated in these cells. An HSV-1-derived oncolytic mutant, R3616, was also found to infect preferentially both transformed cells and PKR-/- (but not PKR+/+) mouse embryo fibroblasts. These observations suggest that HSV-1 specifically targets cells with an activated Ras signalling pathway, and have important ramifications in the use of engineered HSV in cancer therapy, the development of strategies against HSV infections, and the controversial role of HSV in human cancers. PMID- 11483961 TI - Nicastrin binds to membrane-tethered Notch. AB - The presenilins and nicastrin, a type 1 transmembrane glycoprotein, form high molecular weight complexes that are involved in cleaving the beta-amyloid precursor protein (betaAPP) and Notch in their transmembrane domains. The former process (termed gamma-secretase cleavage) generates amyloid beta-peptide (Abeta), which is involved in the pathogenesis of Alzheimer's disease. The latter process (termed S3-site cleavage) generates Notch intracellular domain (NICD), which is involved in intercellular signalling. Nicastrin binds both full-length betaAPP and the substrates of gamma-secretase (C99- and C83-betaAPP fragments), and modulates the activity of gamma-secretase. Although absence of the Caenorhabditis elegans nicastrin homologue (aph-2) is known to cause an embryonic-lethal glp-1 phenotype, the role of nicastrin in this process has not been explored. Here we report that nicastrin binds to membrane-tethered forms of Notch (substrates for S3-site cleavage of Notch), and that, although mutations in the conserved 312-369 domain of nicastrin strongly modulate gamma-secretase, they only weakly modulate the S3-site cleavage of Notch. Thus, nicastrin has a similar role in processing Notch and betaAPP, but the 312-369 domain may have differential effects on these activities. In addition, we report that the Notch and betaAPP pathways do not significantly compete with each other. PMID- 11483962 TI - The Eps15 C. elegans homologue EHS-1 is implicated in synaptic vesicle recycling. AB - Eps15 represents the prototype of a family of evolutionarily conserved proteins that are characterized by the presence of the EH domain, a protein-protein interaction module, and that are involved in many aspects of intracellular vesicular sorting. Although biochemical and functional studies have implicated Eps15 in endocytosis, its function in the endocytic machinery remains unclear. Here we show that the Caenorhabditis elegans gene, zk1248.3 (ehs-1), is the orthologue of Eps15 in nematodes, and that its product, EHS-1, localizes to synaptic-rich regions. ehs-1-impaired worms showed temperature-dependent depletion of synaptic vesicles and uncoordinated movement. These phenotypes could be correlated with a presynaptic defect in neurotransmission. Impairment of EHS-1 function in dyn-1(ky51) worms, which express a mutant form of dynamin and display a temperature-sensitive locomotion defect, resulted in a worsening of the dyn-1 phenotype and uncoordination at the permissive temperature. Thus, ehs-1 and dyn-1 interact genetically. Moreover, mammalian Eps15 and dynamin protein were shown to interact in vivo. Taken together, our results indicate that EHS-1 acts in synaptic vesicle recycling and that its function might be linked to that of dynamin. PMID- 11483963 TI - Mammalian STAG3 is a cohesin specific to sister chromatid arms in meiosis I. AB - Cohesins, which have been characterized in budding yeast and Xenopus, are multisubunit protein complexes involved in sister chromatid cohesion. Regulation of the interactions among different cohesin subunits and the assembly/disassembly of the cohesin complex to chromatin are key steps in chromosome segregation. We previously characterized the mammalian STAG3 protein as a component of the synaptonemal complex that is specifically expressed in germinal cells, although its function in meiosis remains unknown. Here we show that STAG3 has a role in sister chromatid arm cohesion during mammalian meiosis I. Immunofluorescence results in prophase I cells suggest that STAG3 is a component of the axial/lateral element of the synaptonemal complex. In metaphase I, STAG3 is located at the interchromatid domain and is absent from the chiasma region. In late anaphase I and the later stages of meiosis, STAG3 is not detected. STAG3 interacts with the structural maintenance chromosome proteins SMC1 and SMC3, which have been reported to be subunits of the mitotic cohesin complex. We propose that STAG3 is a sister chromatid arm cohesin that is specific to mammalian meiosis I. PMID- 11483964 TI - Reproducible but dynamic positioning of DNA in chromosomes during mitosis. AB - How DNA is folded into chromosomes is unknown. Mitotic chromosome banding shows reproducibility in longitudinal compaction at a resolution of several megabase pairs, but it is less clear whether DNA sequences are targeted laterally to specific locations. The in vitro chromosome assembly of prokaryotic DNA suggests that there is a lack of sequence requirements for chromosome condensation, implying an absence of DNA targeting. Protein extraction experiments indicate, however, that specific DNA sequences may bind to a chromosome scaffold. Chromosome banding patterns, using dyes with differential sequence specificity, have been interpreted to result from the alignment of AT-rich sequences in a partially helically folded chromosome scaffold. But fluorescence in situ hybridization experiments, perhaps owing to technical limitations, have shown at best only slight deviation from a random, lateral sequence distribution. Here we show that there is highly reproducible targeting of specific chromosome segments to the metaphase chromatid axis, but that these segments localize to the periphery of prophase and telophase chromosomes. Unfolding intermediates during anaphase and telophase suggest that sequence repositioning occurs through the global uncoiling of an underlying chromatid structure. PMID- 11483965 TI - Deep impact? PMID- 11483966 TI - Which Ras rides the raft? PMID- 11483968 TI - Does endo-symbiosis explain the origin of the nucleus? PMID- 11483969 TI - Does endo-symbiosis explain the origin of the nucleus? PMID- 11483971 TI - DNA microarrays and beyond: completing the journey from tissue to cell. AB - For the cell biologist, identifying changes in gene expression using DNA microarrays is just the start of a long journey from tissue to cell. We discuss how chip users can first filter noise (false-positives) from daunting microarray datasets. Combining laser capture microdissection with real-time polymerase chain reaction and reverse transcription is a helpful follow-up step that allows expression of selected genes to be quantified using sensitive new in situ hybridization and immunohistochemical methods based on tyramide signal amplification. PMID- 11483972 TI - PX domains: attracted by phosphoinositides. PMID- 11483973 TI - Getting activated with poly-ubiquitination. PMID- 11483974 TI - The origin of CDK regulation. PMID- 11483975 TI - All for one and one for all. PMID- 11483976 TI - Signalling and endocytosis: Wnt breaks down on back roads. PMID- 11483980 TI - Navigating gene expression using microarrays--a technology review. AB - Parallel quantification of large numbers of messenger RNA transcripts using microarray technology promises to provide detailed insight into cellular processes involved in the regulation of gene expression. This should allow new understanding of signalling networks that operate in the cell and of the molecular basis and classification of disease. But can the technology deliver such far-reaching promises? PMID- 11483981 TI - Greasing the wheels of secretory transport. AB - Ever since George Palade's pioneering studies of zymogen secretion from pancreatic acinar cells, the underlying molecular mechanisms of vesicle-mediated protein transport have captivated cell biologists and biochemists. A watershed meeting on "Phosphoinositides and the Golgi", held at the Fogarty International Center of the National Institutes of Health in Bethesda, Maryland (March 13-14, 2001), provided reinterpretation and striking new insights about the functions of this phospholipid class in intracellular protein trafficking. PMID- 11483982 TI - Strategies for the systematic sequencing of complex genomes. AB - Recent spectacular advances in the technologies and strategies for DNA sequencing have profoundly accelerated the detailed analysis of genomes from myriad organisms. The past few years alone have seen the publication of near-complete or draft versions of the genome sequence of several well-studied, multicellular organisms - most notably, the human. As well as providing data of fundamental biological significance, these landmark accomplishments have yielded important strategic insights that are guiding current and future genome-sequencing projects. PMID- 11483983 TI - Determining centromere identity: cyclical stories and forking paths. AB - The centromere is the genetic locus required for chromosome segregation. It is the site of spindle attachment to the chromosomes and is crucial for the transfer of genetic information between cell and organismal generations. Although the centromere was first recognized more than 120 years ago, little is known about what determines its site(s) of activity, and how it contributes to kinetochore formation and spindle attachment. Recent work in this field has supported the hypothesis that most eukaryotic centromeres are determined epigenetically rather than by primary DNA sequence. Here, we review recent studies that have elucidated the organization and functions of centromeric chromatin, and evaluate present-day models for how centromere identity and propagation are determined. PMID- 11483984 TI - The role of selfish genetic elements in eukaryotic evolution. AB - 'Selfish genetic elements', such as transposons, homing endonucleases, meiotic drive chromosomes and heritable microorganisms, are common features of eukaryotes. However, their importance in the evolution of eukaryotic genomes is still controversial. In this review, we discuss these diverse elements and their potential importance in the evolution of genetic systems, adaptation, and the extinction and birth of species. PMID- 11483985 TI - Plant evolution and development in a post-genomic context. AB - Large-scale gene-sequencing projects that have been undertaken in animals have involved organisms from contrasting taxonomic groups, such as worm, fly and mammal. By contrast, similar botanical projects have focused exclusively on flowering plants. This has made it difficult to carry out fundamental research on how plants have evolved from simple to complex forms - a task that has been very successful in animals. However, in the flowering plants, the many completely or partially sequenced genomes now becoming available will provide a powerful tool to investigate the details of evolution in one group of related organisms. PMID- 11483986 TI - Genetics of morphogen gradients. AB - The organization of cells and tissues is controlled by the action of 'form giving' signalling molecules, or morphogens, which pattern a developmental field in a concentration-dependent manner. As the fate of each cell in the field depends on the level of the morphogen signal, the concentration gradient of the morphogen prefigures the pattern of development. In recent years, molecular genetic studies in Drosophila melanogaster have allowed tremendous progress in understanding how morphogen gradients are formed and maintained, and the mechanism by which receiving cells respond to the gradient. PMID- 11483987 TI - Genetics of susceptibility to tuberculosis: Mengele's experiments in Auschwitz. AB - Great experiments will always be remembered. I highlight an experiment that was conducted during the Nazi regime in Germany. Not only did the experiment fail, it was also linked to fraud and crimes against humanity. This failed experiment will never be forgotten. PMID- 11483988 TI - Do bacteria have sex? AB - Do bacteria have genes for genetic exchange? The idea that the bacterial processes that cause genetic exchange exist because of natural selection for this process is shared by almost all microbiologists and population geneticists. However, this assumption has been perpetuated by generations of biology, microbiology and genetics textbooks without ever being critically examined. PMID- 11483989 TI - DNA replication origins: from sequence specificity to epigenetics. AB - Site-specific initiation of DNA replication is a conserved function in all organisms. In Escherichia coli and Saccharomyces cerevisiae, DNA replication origins are sequence specific, but in multicellular organisms, origins are not so clearly defined. In this article, I present a model of origin specification by epigenetic mechanisms that allows the establishment of stable chromatin domains, which are characterized by autonomous replication. According to this model, origins of DNA replication help to establish domains of gene expression for the generation of cell diversity. PMID- 11483990 TI - Mycobacterium tuberculosis: here today, and here tomorrow. AB - Mycobacterium tuberculosis is a highly successful pathogen that parasitizes the macrophages of its host. Its success can be attributed directly to its ability to manipulate the phagosome that it resides in and to prevent the normal maturation of this organelle into an acidic, hydrolytic compartment. As the macrophage is key to clearing the infection, the interplay between the pathogen and its host cell reflects a constant battle for control. PMID- 11483991 TI - Pathogenic trickery: deception of host cell processes. AB - Microbial pathogens cause a spectrum of diseases in humans. Although the disease mechanisms vary considerably, most pathogens have developed virulence factors that interact with host molecules, often usurping normal cellular processes, including cytoskeletal dynamics and vesicle targeting. These virulence factors often mimic host molecules, and mediate events as diverse as bacterial invasion, antiphagocytosis, and intracellular parastism. PMID- 11483992 TI - Four deaths and a funeral: from caspases to alternative mechanisms. AB - A single family of proteases, the caspases, has long been considered the pivotal executioner of all programmed cell death. However, recent findings of evolutionarily conserved, caspase-independent controlled death mechanisms have opened new perspectives on the biology of cell demise, with particular implications for neurobiology, cancer research and immunological processes. PMID- 11483993 TI - Transcriptional regulation by the phosphorylation-dependent factor CREB. AB - The transcription factor CREB -- for 'cyclic AMP response element-binding protein' -- functions in glucose homeostasis, growth-factor-dependent cell survival, and has been implicated in learning and memory. CREB is phosphorylated in response to various signals, but how is specificity achieved in these signalling pathways? PMID- 11483994 TI - The kamikaze approach to membrane transport. AB - Membrane transport proteins catalyse the movement of molecules into and out of cells and organelles, but their hydrophobic and metastable nature often makes them difficult to study by traditional means. Novel approaches that have been developed and applied to one membrane transport protein, the lactose permease from Escherichia coli, are now being used to study various other membrane proteins. PMID- 11483995 TI - A bouquet makes ends meet. AB - The 'chromosomal bouquet' is a polarized chromosomal arrangement that is highly conserved among eukaryotes. There have been many hypotheses about its role in the pairing of meiotic chromosomes, but until recently these have been difficult to test. PMID- 11483996 TI - The phosphatidylserine receptor: a crucial molecular switch? AB - The uptake and removal of necrotic or lysed cells involves inflammation and an immune response, due in part to processes that involve members of the collectin family, surface calreticulin and CD91. Clearance of apoptotic cells, by contrast, does not induce either inflammation or immunity. Could the phosphatidylserine receptor be the molecular switch that determines what the outcome will be? PMID- 11483998 TI - Ghrelin: an orexigenic and somatotrophic signal from the stomach. PMID- 11483999 TI - From the perception of action to the understanding of intention. PMID- 11483997 TI - Correlated neuronal activity and the flow of neural information. PMID- 11484000 TI - New insights into the functions of the superior temporal cortex. PMID- 11484001 TI - Pathophysiology of levodopa-induced dyskinesia: potential for new therapies. AB - Involuntary movements--or dyskinesias--are a debilitating complication of levodopa therapy for Parkinson's disease, and is experienced in most patients. Despite the importance of this problem, little was known about the cause of dyskinesia until recently; however, this situation has changed significantly in the past few years. Our increased understanding of levodopa-induced dyskinesia is not only valuable for improving patient care, but also in providing us with new insights into the functional organization of the basal ganglia and motor systems. PMID- 11484002 TI - Failure of the ubiquitin-proteasome system in Parkinson's disease. PMID- 11484004 TI - How not to make friends. PMID- 11484003 TI - Alzheimer's disease and Abeta toxicity: from top to bottom. PMID- 11484005 TI - Time for orbiting lab to find true purpose. PMID- 11484006 TI - Tough decisions loom as funding crisis hits space-station research. PMID- 11484008 TI - Johns Hopkins embroiled in fresh misconduct allegations. PMID- 11484009 TI - Review blames BSE outbreak on calf feed. PMID- 11484010 TI - Pressure grows over US blood ban. PMID- 11484011 TI - Hopes of biotech interest spur Latvian population genetics. PMID- 11484014 TI - Public library set to turn publisher as boycott looms. PMID- 11484017 TI - When the going gets tough... PMID- 11484018 TI - Measuring the immeasurable. PMID- 11484019 TI - Vets asked valuable questions about foot-and-mouth measures. PMID- 11484020 TI - Always a role for debate between disciplines. PMID- 11484021 TI - Beneficiaries should pay. PMID- 11484022 TI - Research doesn't denigrate humanity. PMID- 11484028 TI - At cross purposes. PMID- 11484029 TI - Climbing life's tree. PMID- 11484030 TI - Flexible electronic futures. PMID- 11484031 TI - Demography. Uncertain population forecasts. PMID- 11484032 TI - Nanotechnology. Boning up on biology. PMID- 11484035 TI - Superconductivity. Is kinky conventional? PMID- 11484034 TI - Gene regulation. Cycling silence. PMID- 11484037 TI - Obituary. Viktor Hamburger (1900-2001). PMID- 11484038 TI - Fossil molar from a Madagascan marsupial. PMID- 11484039 TI - Maternal investment. Sex differences in avian yolk hormone levels. AB - It has been suggested that female birds put more resources into eggs fathered by attractive males by laying larger eggs or by adding more testosterone, but this inference could be undermined if eggs of different sex are provisioned differently, as these studies did not control for sex differences. Here we compare hormone concentrations in the yolks of male and female eggs and find that these are significantly different. Our results indicate that it is premature to conclude that female birds invest more in eggs sired by a preferred male, and raise the possibility that yolk sex steroids may be part of the sex-determining process in birds. PMID- 11484041 TI - Ecology. Global amphibian population declines. AB - The decline and disappearance of relatively undisturbed populations of amphibians in several high-altitude regions since the 1970s suggests that they may have suffered a global decline, perhaps with a common cause or causes. Houlahan et al. examined means of trends for 936 amphibian populations and concluded that global declines began in the late 1950s, peaked in the 1960s, and have continued at a reduced rate since. Here we re-analyse their data using a method that accounts for the sampling of different populations over different time periods, and find evidence of a mean global decline in monitored populations only in the 1990s. However it is calculated, the global mean not only masks substantial spatial and temporal variation in population trends and sampling effort, but also fails to distinguish between a global decline with global causes and the cumulative effects of local declines with local causes. PMID- 11484043 TI - The Earth's mantle. AB - Seismological images of the Earth's mantle reveal three distinct changes in velocity structure, at depths of 410, 660 and 2,700 km. The first two are best explained by mineral phase transformations, whereas the third-the D" layer probably reflects a change in chemical composition and thermal structure. Tomographic images of cold slabs in the lower mantle, the displacements of the 410-km and 660-km discontinuities around subduction zones, and the occurrence of small-scale heterogeneities in the lower mantle all indicate that subducted material penetrates the deep mantle, implying whole-mantle convection. In contrast, geochemical analyses of the basaltic products of mantle melting are frequently used to infer that mantle convection is layered, with the deeper mantle largely isolated from the upper mantle. We show that geochemical, seismological and heat-flow data are all consistent with whole-mantle convection provided that the observed heterogeneities are remnants of recycled oceanic and continental crust that make up about 16 and 0.3 per cent, respectively, of mantle volume. PMID- 11484044 TI - Ground-based observation of emission lines from the corona of a red-dwarf star. AB - All 'solar-like' stars are surrounded by coronae, which contain magnetically confined plasma at temperatures above 106 K. (Until now, only the Sun's corona could be observed in the optical-as a shimmering envelope during a total solar eclipse.) As the underlying stellar 'surfaces'-the photospheres-are much cooler, some non-radiative process must be responsible for heating the coronae. The heating mechanism is generally thought to be magnetic in origin, but is not yet understood even for the case of the Sun. Ultraviolet emission lines first led to the discovery of the enormous temperature of the Sun's corona, but thermal emission from the coronae of other stars has hitherto been detectable only from space, at X-ray wavelengths. Here we report the detection of emission from highly ionized iron (Fe XIII at 3,388.1 A) in the corona of the red-dwarf star CN Leonis, using a ground-based telescope. The X-ray flux inferred from our data is consistent with previously measured X-ray fluxes, and the non-thermal line width of 18.4 km s-1 indicates great similarities between solar and stellar coronal heating mechanisms. The accessibility and spectral resolution (45,000) of the ground-based instrument are much better than those of X-ray satellites, so a new window to the study of stellar coronae has been opened. PMID- 11484045 TI - Evidence for ubiquitous strong electron-phonon coupling in high-temperature superconductors. AB - Coupling between electrons and phonons (lattice vibrations) drives the formation of the electron pairs responsible for conventional superconductivity. The lack of direct evidence for electron-phonon coupling in the electron dynamics of the high transition-temperature superconductors has driven an intensive search for an alternative mechanism. A coupling of an electron with a phonon would result in an abrupt change of its velocity and scattering rate near the phonon energy. Here we use angle-resolved photoemission spectroscopy to probe electron dynamics-velocity and scattering rate-for three different families of copper oxide superconductors. We see in all of these materials an abrupt change of electron velocity at 50-80 meV, which we cannot explain by any known process other than to invoke coupling with the phonons associated with the movement of the oxygen atoms. This suggests that electron-phonon coupling strongly influences the electron dynamics in the high-temperature superconductors, and must therefore be included in any microscopic theory of superconductivity. PMID- 11484046 TI - Fragile-to-strong transition and polyamorphism in the energy landscape of liquid silica. AB - Liquid silica is the archetypal glass former, and compounds based on silica are ubiquitous as natural and man-made amorphous materials. Liquid silica is also the extreme case of a 'strong' liquid, in that the variation of viscosity with temperature closely follows the Arrhenius law as the liquid is cooled toward its glass transition temperature. In contrast, most liquids are to some degree 'fragile', showing significantly faster increases in their viscosity as the glass transition temperature is approached. Recent studies have demonstrated the controlling influence of the potential energy hypersurface (or 'energy landscape') of the liquid on the transport properties near the glass transition. But the origin of strong liquid behaviour in terms of the energy landscape has not yet been resolved. Here we study the static and dynamic properties of liquid silica over a wide range of temperature and density using computer simulations. The results reveal a change in the energy landscape with decreasing temperature, which underlies a transition from a fragile liquid at high temperature to a strong liquid at low temperature. We also show that a specific heat anomaly is associated with this fragile-to-strong transition, and suggest that this anomaly is related to the polyamorphic behaviour of amorphous solid silica. PMID- 11484047 TI - Growth dynamics of pentacene thin films. AB - The recent demonstration of single-crystal organic optoelectronic devices has received widespread attention. But practical applications of such devices require the use of inexpensive organic films deposited on a wide variety of substrates. Unfortunately, the physical properties of these organic thin films do not compare favourably to those of single-crystal materials. Moreover, the basic physical principles governing organic thin-film growth and crystallization are not well understood. Here we report an in situ study of the evolution of pentacene thin films, utilizing the real-time imaging capabilities of photoelectron emission microscopy. By a combination of careful substrate preparation and surface energy control, we succeed in growing thin films with single-crystal grain sizes approaching 0.1 millimetre (a factor of 20-100 larger than previously achieved), which are large enough to fully contain a complete device. We find that organic thin-film growth closely mimics epitaxial growth of inorganic materials, and we expect that strategies and concepts developed for these inorganic systems will provide guidance for the further development and optimization of molecular thin film devices. PMID- 11484048 TI - Ionic conductivity in crystalline polymer electrolytes. AB - Polymer electrolytes are the subject of intensive study, in part because of their potential use as the electrolyte in all-solid-state rechargeable lithium batteries. These materials are formed by dissolving a salt (for example LiI) in a solid host polymer such as poly(ethylene oxide) (refs 2, 3, 4, 5, 6), and may be prepared as both crystalline and amorphous phases. Conductivity in polymer electrolytes has long been viewed as confined to the amorphous phase above the glass transition temperature, Tg, where polymer chain motion creates a dynamic, disordered environment that plays a critical role in facilitating ion transport. Here we show that, in contrast to this prevailing view, ionic conductivity in the static, ordered environment of the crystalline phase can be greater than that in the equivalent amorphous material above Tg. Moreover, we demonstrate that ion transport in crystalline polymer electrolytes can be dominated by the cations, whereas both ions are generally mobile in the amorphous phase. Restriction of mobility to the lithium cation is advantageous for battery applications. The realization that order can promote ion transport in polymers is interesting in the context of electronically conducting polymers, where crystallinity favours electron transport. PMID- 11484049 TI - Covariation of carbon dioxide and temperature from the Vostok ice core after deuterium-excess correction. AB - Ice-core measurements of carbon dioxide and the deuterium palaeothermometer reveal significant covariation of temperature and atmospheric CO2 concentrations throughout the climate cycles of the past ice ages. This covariation provides compelling evidence that CO2 is an important forcing factor for climate. But this interpretation is challenged by some substantial mismatches of the CO2 and deuterium records, especially during the onset of the last glaciation, about 120 kyr ago. Here we incorporate measurements of deuterium excess from Vostok in the temperature reconstruction and show that much of the mismatch is an artefact caused by variations of climate in the water vapour source regions. Using a model that corrects for this effect, we derive a new estimate for the covariation of CO2 and temperature, of r2 = 0.89 for the past 150 kyr and r2 = 0.84 for the period 350-150 kyr ago. Given the complexity of the biogeochemical systems involved, this close relationship strongly supports the importance of carbon dioxide as a forcing factor of climate. Our results also suggest that the mechanisms responsible for the drawdown of CO2 may be more responsive to temperature than previously thought. PMID- 11484050 TI - Chemical interaction of Fe and Al(2)O3 as a source of heterogeneity at the Earth's core-mantle boundary. AB - Seismological studies have revealed that a complex texture or heterogeneity exists in the Earth's inner core and at the boundary between core and mantle. These studies highlight the importance of understanding the properties of iron when modelling the composition and dynamics of the core and the interaction of the core with the lowermost mantle. One of the main problems in inferring the composition of the lowermost mantle is our lack of knowledge of the high-pressure and high-temperature chemical reactions that occur between iron and the complex Mg-Fe-Si-Al-oxides which are thought to form the bulk of the Earth's lower mantle. A number of studies have demonstrated that iron can react with MgSiO3 perovskite at high pressures and high temperatures, and it was proposed that the chemical nature of this process involves the reduction of silicon by the more electropositive iron. Here we present a study of the interaction between iron and corundum (Al(2)O3) in electrically- and laser-heated diamond anvil cells at 2,000 2,200 K and pressures up to 70 GPa, simulating conditions in the Earth's deep interior. We found that at pressures above 60 GPa and temperatures of 2,200 K, iron and corundum react to form iron oxide and an iron-aluminium alloy. Our results demonstrate that iron is able to reduce aluminium out of oxides at core mantle boundary conditions, which could provide an additional source of light elements in the Earth's core and produce significant heterogeneity at the core mantle boundary. PMID- 11484051 TI - The last of the dinosaur titans: a new sauropod from Madagascar. AB - The Titanosauria, the last surviving group of the giant sauropod dinosaurs, attained a near-global distribution by the close of the Cretaceous period (65 Myr ago). With the exception of a few new discoveries in Argentina, most titanosaurs are known only from fragmentary postcranial skeletons and rare, isolated skull elements. Here we describe the most complete titanosaur yet discovered. Rapetosaurus krausei gen. et sp. nov., from the Maevarano Formation of Madagascar, provides a view of titanosaur anatomy from head to tail. A total evidence phylogenetic analysis supports a close relationship between brachiosaurids and titanosaurs (Titanosauriformes). The inclusion of cranial data from Rapetosaurus also lays to rest questions concerning the phylogeny of the enigmatic Mongolian genera Nemegtosaurus and Quaesitosaurus. In spite of their elongated, diplodocoid-like skulls, all three taxa are now firmly nested within Titanosauria. PMID- 11484052 TI - Neanderthal cranial ontogeny and its implications for late hominid diversity. AB - Homo neanderthalensis has a unique combination of craniofacial features that are distinct from fossil and extant 'anatomically modern' Homo sapiens (modern humans). Morphological evidence, direct isotopic dates and fossil mitochondrial DNA from three Neanderthals indicate that the Neanderthals were a separate evolutionary lineage for at least 500,000 yr. However, it is unknown when and how Neanderthal craniofacial autapomorphies (unique, derived characters) emerged during ontogeny. Here we use computerized fossil reconstruction and geometric morphometrics to show that characteristic differences in cranial and mandibular shape between Neanderthals and modern humans arose very early during development, possibly prenatally, and were maintained throughout postnatal ontogeny. Postnatal differences in cranial ontogeny between the two taxa are characterized primarily by heterochronic modifications of a common spatial pattern of development. Evidence for early ontogenetic divergence together with evolutionary stasis of taxon-specific patterns of ontogeny is consistent with separation of Neanderthals and modern humans at the species level. PMID- 11484053 TI - Habitat structure and population persistence in an experimental community. AB - Understanding spatial population dynamics is fundamental for many questions in ecology and conservation. Many theoretical mechanisms have been proposed whereby spatial structure can promote population persistence, in particular for exploiter victim systems (host-parasite/pathogen, predator-prey) whose interactions are inherently oscillatory and therefore prone to extinction of local populations. Experiments have confirmed that spatial structure can extend persistence, but it has rarely been possible to identify the specific mechanisms involved. Here we use a model-based approach to identify the effects of spatial population processes in experimental systems of bean plants (Phaseolus lunatus), herbivorous mites (Tetranychus urticae) and predatory mites (Phytoseiulus persimilis). On isolated plants, and in a spatially undivided experimental system of 90 plants, prey and predator populations collapsed; however, introducing habitat structure allowed long-term persistence. Using mechanistic models, we determine that spatial population structure did not contribute to persistence, and spatially explicit models are not needed. Rather, habitat structure reduced the success of predators at locating prey outbreaks, allowing between-plant asynchrony of local population cycles due to random colonization events. PMID- 11484054 TI - The end of world population growth. AB - There has been enormous concern about the consequences of human population growth for the environment and for social and economic development. But this growth is likely to come to an end in the foreseeable future. Improving on earlier methods of probabilistic forecasting, here we show that there is around an 85 per cent chance that the world's population will stop growing before the end of the century. There is a 60 per cent probability that the world's population will not exceed 10 billion people before 2100, and around a 15 per cent probability that the world's population at the end of the century will be lower than it is today. For different regions, the date and size of the peak population will vary considerably. PMID- 11484055 TI - Retrospective and prospective coding for predicted reward in the sensory thalamus. AB - Reward is important for shaping goal-directed behaviour. After stimulus-reward associative learning, an organism can assess the motivational value of the incoming stimuli on the basis of past experience (retrospective processing), and predict forthcoming rewarding events (prospective processing). The traditional role of the sensory thalamus is to relay current sensory information to cortex. Here we find that non-primary thalamic neurons respond to reward-related events in two ways. The early, phasic responses occurred shortly after the onset of the stimuli and depended on the sensory modality. Their magnitudes resisted extinction and correlated with the learning experience. The late responses gradually increased during the cue and delay periods, and peaked just before delivery of the reward. These responses were independent of sensory modality and were modulated by the value and timing of the reward. These observations provide new evidence that single thalamic neurons can code for the acquired significance of sensory stimuli in the early responses (retrospective coding) and predict upcoming reward value in the late responses (prospective coding). PMID- 11484056 TI - Practising orientation identification improves orientation coding in V1 neurons. AB - The adult brain shows remarkable plasticity, as demonstrated by the improvement in fine sensorial discriminations after intensive practice. The behavioural aspects of such perceptual learning are well documented, especially in the visual system. Specificity for stimulus attributes clearly implicates an early cortical site, where receptive fields retain fine selectivity for these attributes; however, the neuronal correlates of a simple visual discrimination task remained unidentified. Here we report electrophysiological correlates in the primary visual cortex (V1) of monkeys for learning orientation identification. We link the behavioural improvement in this type of learning to an improved neuronal performance of trained compared to naive neurons. Improved long-term neuronal performance resulted from changes in the characteristics of orientation tuning of individual neurons. More particularly, the slope of the orientation tuning curve that was measured at the trained orientation increased only for the subgroup of trained neurons most likely to code the orientation identified by the monkey. No modifications of the tuning curve were observed for orientations for which the monkey had not been trained. Thus training induces a specific and efficient increase in neuronal sensitivity in V1. PMID- 11484057 TI - Regulation of DNA replication fork progression through damaged DNA by the Mec1/Rad53 checkpoint. AB - The checkpoint kinase proteins Mec1 and Rad53 are required in the budding yeast, Saccharomyces cerevisiae, to maintain cell viability in the presence of drugs causing damage to DNA or arrest of DNA replication forks. It is thought that they act by inhibiting cell cycle progression, allowing time for DNA repair to take place. Mec1 and Rad53 also slow S phase progression in response to DNA alkylation, although the mechanism for this and its relative importance in protecting cells from DNA damage have not been determined. Here we show that the DNA-alkylating agent methyl methanesulphonate (MMS) profoundly reduces the rate of DNA replication fork progression; however, this moderation does not require Rad53 or Mec1. The accelerated S phase in checkpoint mutants, therefore, is primarily a consequence of inappropriate initiation events. Wild-type cells ultimately complete DNA replication in the presence of MMS. In contrast, replication forks in checkpoint mutants collapse irreversibly at high rates. Moreover, the cytotoxicity of MMS in checkpoint mutants occurs specifically when cells are allowed to enter S phase with DNA damage. Thus, preventing damage induced DNA replication fork catastrophe seems to be a primary mechanism by which checkpoints preserve viability in the face of DNA alkylation. PMID- 11484058 TI - The DNA replication checkpoint response stabilizes stalled replication forks. AB - In response to DNA damage and blocks to replication, eukaryotes activate the checkpoint pathways that prevent genomic instability and cancer by coordinating cell cycle progression with DNA repair. In budding yeast, the checkpoint response requires the Mec1-dependent activation of the Rad53 protein kinase. Active Rad53 slows DNA synthesis when DNA is damaged and prevents firing of late origins of replication. Further, rad53 mutants are unable to recover from a replication block. Mec1 and Rad53 also modulate the phosphorylation state of different DNA replication and repair enzymes. Little is known of the mechanisms by which checkpoint pathways interact with the replication apparatus when DNA is damaged or replication blocked. We used the two-dimensional gel technique to examine replication intermediates in response to hydroxyurea-induced replication blocks. Here we show that hydroxyurea-treated rad53 mutants accumulate unusual DNA structures at replication forks. The persistence of these abnormal molecules during recovery from the hydroxyurea block correlates with the inability to dephosphorylate Rad53. Further, Rad53 is required to properly maintain stable replication forks during the block. We propose that Rad53 prevents collapse of the fork when replication pauses. PMID- 11484059 TI - Rb targets histone H3 methylation and HP1 to promoters. AB - In eukaryotic cells the histone methylase SUV39H1 and the methyl-lysine binding protein HP1 functionally interact to repress transcription at heterochromatic sites. Lysine 9 of histone H3 is methylated by SUV39H1 (ref. 2), creating a binding site for the chromo domain of HP1 (refs 3, 4). Here we show that SUV39H1 and HP1 are both involved in the repressive functions of the retinoblastoma (Rb) protein. Rb associates with SUV39H1 and HP1 in vivo by means of its pocket domain. SUV39H1 cooperates with Rb to repress the cyclin E promoter, and in fibroblasts that are disrupted for SUV39, the activity of the cyclin E and cyclin A2 genes are specifically elevated. Chromatin immunoprecipitations show that Rb is necessary to direct methylation of histone H3, and is necessary for binding of HP1 to the cyclin E promoter. These results indicate that the SUV39H1-HP1 complex is not only involved in heterochromatic silencing but also has a role in repression of euchromatic genes by Rb and perhaps other co-repressor proteins. PMID- 11484064 TI - Pharmacology of the thromboxane receptor antagonist and thromboxane synthase inhibitor BM-531. AB - BM-531 (N-tert-butyl-N'-[(2-cyclohexylamino-5-nitrobenzene)sulfonyl]urea), a torasemide derivative, is a novel noncarboxylic thromboxane receptor antagonist and thromboxane synthase inhibitor. Indeed, its affinity for human washed platelet TXA2 receptors labeled with [3H]SQ-29548 (IC50 = 0.0078 microM) is higher than sulotroban (IC50 = 0.93 microM) and SQ-29548 (IC50 = 0.021 microM). Moreover, BM-531 is characterized by a potent antiaggregatory property. Indeed, on one hand, in human citrated platelet-rich plasma BM-531 prevents platelet aggregation induced by arachidonic acid (600 microM) (ED100 = 0.125 microM), U 46619, a stable TXA2 agonist (1 microM) (ED50 = 0.482 microM) or collagen (1 microgram/mL) (percentage of inhibition: 42.9% at 10 microM) and inhibits the second wave of ADP (2 microM)-induced aggregation. On the other hand, when BM-531 is incubated in whole blood from healthy donors, the closure time measured by the recently developed platelet function analyser (PFA-100) is significantly prolonged. In addition, at the concentrations of 10 and 1 microM, BM-531 totally prevents the production of TXB2 by human platelets activated by arachidonic acid. Finally, at 10 microM, BM-531 significantly prevents rat fundus contractions induced by U-46619 but not by prostacyclin. These results suggest that BM-531, which is devoid of the diuretic property of torasemide, can be regarded as a promising antiplatelet agent. PMID- 11484065 TI - Ifetroban sodium: an effective TxA2/PGH2 receptor antagonist. AB - This review presents a comprehensive discussion on the chemistry, pharmacokinetics, and pharmacodynamics of ifetroban sodium, a new thomboxane A2/prostaglandin H2 receptor antagonist. Thromboxane A2 is an arachidonic acid product, formed by the enzyme cyclooxygenase. In contrast to other cyclooxygenase products, thromboxane A2 has been shown to be involved in vascular contraction and has been implicated in platelet activation. In general, results of clinical studies and animal experiments indicate that hypertension is associated with hyperaggregability of platelets and increased thomboxane A2 levels in blood, urine, and tissues. The precursors to thromboxane A2, prostaglandin G2, and prostaglandin H2, also bind and activate the same receptors. Thus, a receptor antagonist was thought to be an improved strategy for reversing the actions of thromboxane A2/prostaglandin H2, rather than a thromboxane synthesis inhibitor. This review describes new methods for the synthesis and analysis of ifetroban, its tissue distribution, and its actions in a variety of animal models and disease states. We describe studies on the mechanisms of how ifetroban relaxes experimentally contracted isolated vascular tissue, and on the effects of ifetroban on myocardial ischemia, hypertension, stroke, thrombosis, and its effects on platelets. These experiments were conducted on several animal models, including dog, ferret, and rat, as well as on humans. Clinical studies are also described. These investigations show that ifetroban sodium is effective at reversing the effects of thromboxane A2- and prostaglandin H2-mediated processes. PMID- 11484066 TI - Xemilofiban/orbofiban: insight into drug development. AB - A number of studies have reported on the successful use of intravenous glycoprotein IIb/IIIa receptor antagonists in patients with unstable angina or undergoing percutaneous interventions. The promise of interrupting the aggregation of platelets in the setting of unstable plaques on a chronic basis had led to the evaluation of several oral agents for longer-term administration. The development program of two of these agents, xemilofiban and orbofiban, will be reviewed and evaluated to understand the selection process of therapeutic targets for use based upon complex pharmacokinetic and pharmacodynamic responses. A review of the pivotal phase III trial results will also be provided along with insights into the potential reasons for the lack of significant benefit shown with these agents to date. PMID- 11484067 TI - Multilateral in vivo and in vitro protective effects of the novel heat shock protein coinducer, bimoclomol: results of preclinical studies. AB - Bimoclomol, the recently developed non-toxic heat shock protein (HSP) coinducer, was shown to display multilateral protective activities against various forms of stress or injuries at the level of the cell, tissue or organism. The compound enhanced the transcription, translation and expression of the 70 kD heat shock protein (HSP-70) in myogenic and HeLa cell lines exposed to heat stress, and increased cell survival on exposure to otherwise lethal thermal injury. Bimoclomol increased contractility of the working mammalian heart, this effect was associated with the increased intracellular calcium transients due to increased probability of opening of ryanodine receptors in the sarcoplasmic reticulum (SR). In healthy tissues these cardiac effects were evident only at relatively high concentrations of the drug, while in the ischemic myocardium bimoclomol exerted significant cardioprotective and antiarrhythmic effects at submicromolar concentrations. It decreased ischemia-induced reduction of contractility and of cardiac output, and dramatically decreased the elevation of the ST-segment during ischemia as well as the occurrence of ventricular fibrillation upon reperfusion. Bimoclomol was also active in various pathological animal models subjected to acute or chronic stress. In the spontaneously hypertensive rats chronic pretreatment with bimoclomol restored sensitivity of aortic rings to acetylcholine; this effect was accompanied by accumulation of HSP 70 in the tissues. Bimoclomol pretreatment significantly diminished the consequences of vascular disorders associated with diabetes mellitus. Diabetic neuropathy, retinopathy, and nephropathy were prevented or diminished, while wound healing was enhanced by bimoclomol. Enhancement of wound healing by bimoclomol was observed after thermal injury as well as following ultraviolet (UV) irradiation. In addition to the beneficial effects on peripheral angiopathies, bimoclomol antagonized the increase in permeability of blood-brain barrier induced by subarachnoid hemorrhager or arachidonic acid. A general and very important feature of the above effects of bimoclomol was that the drug failed to cause alterations under physiological conditions (except the enhanced calcium release from cardiac sarcoplasmic reticulum). Bimoclomol was effective only under conditions of stress. Consistent with its HSP-coinducer property, bimoclomol alone had very little effect on HSP production. Its protective activity became apparent only in the presence of cell damage. Currently, bimoclomol reached the end of the Phase II clinical trial in a group of 410 patients with diabetic complications. Results of this trial will answer the question, whether a compound with promising in vitro and in vivo preclinical findings will produce the anticipated beneficial effects in humans. In the event of a positive outcome of this trial, the indications for bimoclomol will be substantially extended. PMID- 11484068 TI - Carvedilol: molecular and cellular basis for its multifaceted therapeutic potential. AB - Carvedilol is a unique cardiovascular drug of multifaceted therapeutic potential. Its major molecular targets recognized to date are membrane adrenoceptors (beta 1, beta 2, and alpha 1), reactive oxygen species, and ion channels (K+ and Ca2+). Carvedilol provides prominent hemodynamic benefits mainly through a balanced adrenoceptor blockade, which causes a reduction in cardiac work in association with peripheral vasodilation. This drug assures remarkable cardiovascular protection through its antiproliferative/atherogenic, antiischemic, antihypertrophic, and antiarrhythmic actions. These actions are a consequence of its potent antioxidant effects, amelioration of glucose/lipid metabolism, modulation of neurohumoral factors, and modulation of cardiac electrophysiologic properties. The usefulness of carvedilol in the treatment of hypertension, ischemic heart disease, and congestive heart failure is based on a combination of hemodynamic benefits and cardiovascular protection. PMID- 11484069 TI - Sotalol: the mechanism of its antiarrhythmic-defibrillating effect. AB - This minireview deals with the role of intercellular communication and synchronization in the initiation and maintenance of ventricular fibrillation. It is proposed that myocardial cell junctions might represent a therapeutic substrate for the prevention of this fatal arrhythmia. This hypothesis is supported by the results of recent experimental studies involving elucidation of the mechanism of antiarrhythmic-defibrillating effects of sotalol. Enhancement of intercellular communication and myocardial synchronization are thought to play critical role in the mechanism of action of this drug. PMID- 11484070 TI - The role of impaired early insulin secretion in the pathogenesis of Type II diabetes mellitus. AB - Patients with Type II (non-insulin-dependent) diabetes mellitus manifest abnormalities in insulin action and insulin secretion. It is widely accepted that insulin resistance is an early finding, evident before the onset of hyperglycaemia and predictive of the subsequent development of diabetes. Whether abnormalities in insulin secretion also precede and predict diabetes has been debated. However, recent studies clearly indicate that early insulin secretion plays a critical role in maintaining normal glucose homeostasis. Cross-sectional analyses show that acute insulin secretory responses (AIR) to intravenous glucose are lower in subjects with impaired glucose tolerance and those at high risk for developing diabetes. Prospectively, a low AIR predicts the development of diabetes in several populations. In longitudinal studies, AIR declines dramatically as patients progress from normal to impaired glucose tolerance and ultimately to diabetes. Early insulin secretion is important for the rapid and efficient suppression of endogenous glucose production after a meal. Thus, loss of early insulin secretion initially leads to post-prandial hyperglycaemia which, as the disease progresses, worsens to clinical hyperglycaemia. Strategies that enhance early insulin secretion improve glucose tolerance and represent a novel and more physiologic approach to improving glycaemic control in patients with Type II diabetes mellitus. PMID- 11484071 TI - Uncoupling proteins: functional characteristics and role in the pathogenesis of obesity and Type II diabetes. AB - Uncoupling proteins are mitochondrial carrier proteins which are able to dissipate the proton gradient of the inner mitochondrial membrane. This uncoupling process reduces the amount of ATP generated through an oxidation of fuels. The hypothesis that uncoupling proteins (UCPs) are candidate genes for human obesity or Type II (non-insulin-dependent) diabetes mellitus is based on the finding that a chemical uncoupling of the mitochondrial membrane reduces body adiposity, and that lower metabolic rates predict weight gain. It is straightforward to hypothesize that common polymorphisms of UCP1, UCP2 and UCP3 genes lower metabolic rate by a more efficient energy coupling in the mitochondria. Furthermore, genetically engineered mice over expressing different UCP homologues are lean and resistant to diet-induced obesity. The three uncoupling protein homologue genes UCP1, UCP2, and UCP3 have been investigated for polymorphisms and mutations and their impact on Type II diabetes mellitus, obesity, and body weight gain or BMI. The main conclusion is that variation in the UCP1, UCP2 or UCP3 genes is not associated with major alterations of body weight gain. The contribution of UCP genes towards polygenic obesity and Type II diabetes is evaluated and discussed. PMID- 11484072 TI - alpha-Thalassaemia trait and gestational diabetes mellitus in Hong Kong. AB - AIMS/HYPOTHESIS: The purpose of this study was to examine the association between maternal alpha-thalassaemia trait and the occurrence of gestational diabetes mellitus in at-risk Chinese women in Hong Kong. METHODS: From 3320 pregnant women who had delivered in our hospital and undergone the oral glucose tolerance test for various risk factors over a three-year period, 163 with alpha-thalassaemia trait were identified (study group). The control group consisted of 163 women chosen from the next patient that was matched for maternal age and parity, following each index case. Comparison was made in the incidence of gestational diabetes mellitus defined by the World Health Organisation criteria, obstetric complications, and perinatal outcome. RESULTS: The incidence of gestational diabetes mellitus was higher in the study group (62.0 % vs 14.7 %, p < 0.0001) which had a higher pre-pregnancy body mass index and lower haemoglobin concentrations. Although more patients in the study group had risk factors (41.7 % vs 26.4 %, p = 0.003), there was no difference in the pregnancy outcome or perinatal complications. Among the gestational diabetic women, those with alpha thalassaemia trait were considerably younger and their infants had lower body mass index but there was no significant difference in the outcome. On multiple logistic regression analysis, the alpha-thalassaemia trait remained an important factor in the diagnosis of gestational diabetes (OR 11.74, 95 % CI 6.37-21.63). CONCLUSION/INTERPRETATION: Among women at risk of gestational diabetes, the presence of the alpha-thalassaemia trait is an additional risk factor for gestational diabetes mellitus. PMID- 11484073 TI - Dietary fat and gestational hyperglycaemia. AB - AIMS/HYPOTHESIS: The purpose of this study was to investigate the relation between life-style habits and glucose abnormalities in Caucasian women with and without conventional risk factors for gestational diabetes. METHODS: A total of 126 pregnant women with gestational diabetes, 84 with impaired glucose tolerance and 294 with normal glucose tolerance, identified by sequential screening, were interviewed to determine their usual weekly food pattern, amount of exercise, smoking habits and alcohol intake. RESULTS: Patients with glucose abnormalities were older and shorter in height and had significantly higher BMI before pregnancy, percentage of diabetic first-degree relatives and higher intake of saturated fat. Patients without known risk factors for gestational diabetes (i. e. younger than 35 years of age, BMI < 25 kg/m2, no first-degree diabetic relatives) included 40 with impaired glucose tolerance or gestational diabetes. In a multiple logistic regression model age, short stature, familial diabetes, BMI and percentages of saturated fat were associated with impaired glucose tolerance or gestational diabetes in all patients, after adjustment for gestational age. In patients without conventional risk factors only percentages of saturated fat (OR = 2.0; 95 %-CI = 1.2-3.2) and polyunsaturated fat (OR = 0.85; 95 %-CI = 0.77-0.92) were associated with gestational hyperglycaemia, after adjustment for age, gestational age and BMI. CONCLUSION/INTERPRETATION: Saturated fat has an independent role in the development of gestational glucose abnormalities. This role is more important in the absence of conventional risk factors suggesting that glucose abnormalities could be prevented during pregnancy, at least in some groups of women. PMID- 11484074 TI - Homocysteine-induced inhibition of nitric oxide production in platelets: a study on healthy and diabetic subjects. AB - AIMS/HYPOTHESIS: The molecular mechanisms involved in the platelet activation observed in hyperhomocysteinemia are not known. We aimed to discover if homocysteine concentrations are associated with abnormal platelet nitric oxide production in healthy and diabetic subjects. METHODS: The study cohort included 28 patients with Type I (insulin-dependent) diabetes mellitus, 30 patients with Type II (non-insulin-dependent) diabetes mellitus, and 34 healthy subjects. Homocysteine plasma concentrations were measured by high-performance liquid chromatography. Platelet nitric oxide production was measured using a nitric oxide meter before and after a 3-h incubation with 100 micromol/l homocysteine. Stimulation experiments were done in vitro by the addition of alpha-thrombin (0.2 U/ml). RESULTS: Basal platelet nitric oxide production was lower in diabetic patients than in healthy subjects. Nitric oxide release was reduced by in vitro homocysteine incubation, being lower in platelets from diabetic patients than in platelets from control subjects. Thrombin increased nitric oxide synthesis in platelets from healthy subjects both in the presence and absence of homocysteine. In diabetic subjects thrombin increased nitric oxide release in the absence of homocysteine. But in the presence of homocysteine the response was reduced. An inverse relation was found between plasma homocysteine levels and basal platelet nitric oxide release in diabetic and healthy subjects. CONCLUSION/INTERPRETATION: Homocysteine could exert its atherogenic action in healthy and diabetic subjects partly by inhibiting platelet nitric oxide production with the subsequent increased platelet activation and aggregation. PMID- 11484075 TI - Production and metabolic clearance of glucose under basal conditions in Type II (non-insulin-dependent) diabetes mellitus. AB - AIMS/HYPOTHESIS: The pathogenesis of fasting hyperglycaemia in Type II (non insulin-dependent) diabetes mellitus has yet to be clarified. Rates of glucose production (Ra), utilization and metabolic clearance rate were therefore measured during an extended fast, in control subjects and in Type II diabetic patients. METHODS: Nine subjects with newly-diagnosed or diet-treated diabetes and seven control subjects matched for age and weight (BMI 36.0 +/- 2.4 and 35.3 +/- 3.1 kg/m2 respectively) underwent an overnight fast followed by a 10-h unprimed infusion of [6-3H]glucose. Plasma tracer concentrations were fitted by a single compartment model. RESULTS: The metabolic clearance rate was near-constant [61.7 + 2.4 ml/(min-m2)] in diabetic patients and [75.5 +/- 3.3 ml/(min-m2)] in control subjects (p < 0.05). It was correlated to the glucose concentrations both at t = 0 (r = -0.752, p = 0.0008) and t = 10 h (r = -0.675, p = 0.004). The calculated volume of distribution was 17.3 +/- 1.4 l (18.2 % weight, diabetes), 19.6 +/- 2.4 l (18.4 % weight, control). Glycaemia fell from 10.7 +/- 0.8 mmol/l to 6.5 +/- 0.3 mmol/l by 10 h (p < 0.05) in diabetes and from 5.6 +/- 0.6 to 4.8 +/- 0.1 mmol/l in control subjects (p < 0.05). The rate of glucose production decreased in parallel, from 563 +/- 33 to 363 +/- 23 micromol/(min-m2) (p < 0.05) in diabetes from 419 +/- 20 to 347 +/- 32 micromol/(min-m2) in control subjects. Initial Ra was higher in diabetic patients than in control subjects (p < 0.05) and was highly correlated to glycaemia (r = 0.836, p = 0.0001). By 10 h, Ra had converged in diabetic patients and control subjects and all correlation with glycaemia was lost (r = 0.0017, p = 0.95). CONCLUSIONS/INTERPRETATION: In relatively early diabetes, the more "labile" portion of fasting hyperglycaemia, which subsequently decreased, was closely related to the simultaneously decreasing Ra. The 25 % increase in glucose concentrations which persisted as stabilized Ra, resulted from about a 20 % lower metabolic clearance rate. PMID- 11484076 TI - Insulin-independent and wortmannin-resistant targeting of IRS-3 to the plasma membrane via its pleckstrin homology domain mediates a different interaction with the insulin receptor from that of IRS-1. AB - AIMS/HYPOTHESIS: In primary adipocytes, although IRS-1 and IRS-3 are expressed in comparable amounts, these proteins manifest distinct distribution and significance in insulin signalling. We investigated the molecular basis of the difference between these two proteins. METHODS: In Cos-1 cells transiently expressing rat IRS-1, IRS-3, or chimeric proteins of these two proteins we examined the tyrosine phosphorylation via the wild-type or mutant insulin receptors and evaluated their targeting to the plasma membrane by immunostaining the membrane ghost. RESULTS: In contrast to IRS-1, IRS-3 was tyrosine phosphorylated by the insulin receptor altering Tyr960 to Phe (Y960F), which disrupts the binding site of the PTB domain of IRSs, to an extent comparable to the wild-type receptor. The tyrosine phosphorylation of IRS-3 with the PH domain replacement via the Y960F insulin receptor markedly decreased, whereas that of IRS-3 with the PTB domain alteration was mildly impaired. Insulin-stimulated translocation of IRS-1 to the plasma membrane, as well as that of IRS-3 with the PH domain replacement, was wortmannin-sensitive, although that of IRS-3 was insulin-independent and wortmannin-resistant. CONCLUSIONS/INTERPRETATION: The affinity of the PH domain for the phospholipids in the plasma membrane seems to influence the receptor-substrate interaction required for IRS tyrosine phosphorylation, indicating that the PH domain and the PTB domain of IRSs cooperatively function in insulin-stimulated tyrosine phosphorylation of these proteins. PMID- 11484077 TI - Titre and combination of ICA and autoantibodies to glutamic acid decarboxylase discriminate two clinically distinct types of latent autoimmune diabetes in adults (LADA). AB - ABSTRACT AIMS/HYPOTHESIS: This study aimed to define the immunological parameters which could be used to identify patients with the distinct metabolic features of adult latent autoimmune diabetes. METHODS: Sera of 312 patients with short-term diabetes (duration < 5 years) over 35 years of age at diagnosis were screened for ICA, GAD- and IA2-Ab by antibody assays validated in workshops. The antibody status was correlated with age, BMI, residual beta-cell function, measured by fasting C-peptide, onset of diabetes-related complications and markers of the metabolic syndrome (hypertension and hyperlipidaemia). RESULTS: A total of 51 antibody positive patients were identified. These patients had lower fasting C peptide and less neuropathy and hypertension compared with matched antibody negative patients. However, only patients with two or more antibodies had reduced residual beta-cell function compared with antibody-negative or single antibody positive (ICA or GAD-Ab only) patients. Patients with two or more antibodies were also leaner and had diabetes-related complications or hypertension less frequently than single antibody-positive or antibody negative-patients. IA2 antibody status did not substantially contribute to the diagnosis or differentiation of LADA patients. CONCLUSION/INTERPRETATION: We concluded that the combination of ICA and GAD antibodies and high titre of GAD antibodies are characteristic of patients with insulin deficiency with the clinical features of Type I (insulin-dependent) diabetes mellitus (LADA-type 1). Single antibody positivity and low titre antibodies are markers for LADA-type 2 associated with the clinical and metabolic phenotype of Type II (non-insulin-dependent) diabetes patients. PMID- 11484078 TI - Interleukin-6 and C-reactive protein as early markers of sepsis in patients with diabetic ketoacidosis or hyperosmosis. AB - AIMS/HYPOTHESIS: An early diagnosis of sepsis in patients with diabetic ketoacidosis and hyperosmolar non-ketotic coma is crucial and could save lives. We studied serum C-reactive protein and interleukin-6 to find out how useful these might be for identifying sepsis. METHODS: Sixty one diabetic patients with ketoacidosis or hyperosmolar non-ketotic coma were enrolled. Patients with signs and symptoms of systemic inflammatory response syndrome were identified. Acute phase reactants, including C-reactive protein and interleukin-6, the main cytokine responsible for the induction of acute-phase proteins, were measured on admission and when patients had clinically improved and were euglycaemic. RESULTS: A total of 49 out of 61 patients with diabetic ketoacidosis or hyperosmosis had signs of systemic inflammatory response syndrome. Another 27 patients had systemic inflammatory response syndrome and no signs of infection and 22 patients had systemic inflammatory response syndrome due to proven infection. We detected a significant increase in serum C-reactive protein and interleukin-6 values in patients infected compared with patients not infected with systemic inflammatory response syndrome SIRS. Patients who finally died had much higher levels of these proteins, while there was a prompt reduction of serum C-reactive protein and interleukin-6 early during remission. CONCLUSION/INTERPRETATION: Diabetic ketoacidosis and hyperosmolar non-ketotic coma can often cause a clinical syndrome resembling systemic inflammatory response syndrome. Determination of serum C-reactive protein and interleukin-6 levels is a useful way of excluding an underlying infection early on as well as confirming and monitoring sepsis. PMID- 11484079 TI - Islet amyloid polypeptide promotes beta-cell proliferation in neonatal rat pancreatic islets. AB - AIMS/HYPOTHESIS: We aimed to clarify the role of islet amyloid polypeptide, which is expressed at early embryonic onset, in the proliferation and cell death of neonatal islet cells. METHODS: Fetal islets were prepared from pregnant rats on gestational day 21. Islets were cultured in RPMI 1640 (11.1 mmol/l glucose) + 10 % fetal calf serum (FCS) for 48 h, followed by a 24-h culture period in RPMI 1640 (5.6 mmol/l glucose) + 1 % FCS. The islets were then exposed to rat islet amyloid polypeptide (1-10 nmol/l) for 24 h. RESULTS: Islet amyloid polypeptide increased islet DNA synthesis (dpm/microg of DNA. 6 h) (control 1 % FCS: 3634 +/- 662; 1 nmol/l 6347 +/- 1535; 10 nmol/l 5157 +/- 769; p < 0.05 islet amyloid polypeptide vs control). In accordance with this, a doubling of the autoradiographic labelling index was seen in immunocytochemically stained islet beta cells after exposure to 1 and 10 nmol/l islet amyloid polypeptide. Islet amyloid polypeptide at 1 nmol/l increased the islet insulin content (202 +/- 25 % of control; p < 0.01) and the 24-h medium insulin concentration (1 nmol/l islet amyloid polypeptide: 143 +/- 19 % of control; p < 0.05) but at 10 nmol/l islet amyloid polypeptide these changes did not attain statistical difference. Islet amyloid polypeptide did not have any marked effect on the islet cell death frequency, suggesting that islet amyloid polypeptide is a more potent promoter of proliferation than of programmed cell death. CONCLUSION/INTERPRETATION: Our data indicate islet amyloid polypeptide is a potential regulator of proliferation in neonatal pancreatic islet cells, an effect which can partly be attributed to the proliferation of beta cells. PMID- 11484080 TI - Gliclazide produces high-affinity block of KATP channels in mouse isolated pancreatic beta cells but not rat heart or arterial smooth muscle cells. AB - AIMS/HYPOTHESIS: Sulphonylureas stimulate insulin secretion by closing ATP sensitive potassium (KATP) channels in the pancreatic beta-cell membrane. KATP channels are also found in other tissues, including heart and smooth muscle, where they link cellular metabolism to electrical activity. The sulphonylurea gliclazide blocks recombinant beta-cell KATP channels (Kir6.2/SUR1) but not heart (Kir6.2/SUR2A) or smooth muscle (Kir6.2/SUR2B) KATP channels with high potency. In this study, we examined the specificity of gliclazide for the native (as opposed to recombinant) KATP channels in beta cells, heart and smooth muscle. METHODS: The action of the drug was studied by whole-cell current recordings of native KATP channels in isolated pancreatic beta-cells and myocytes from heart and smooth muscle. RESULTS: Gliclazide blocked whole-cell beta-cell KATP currents with an IC50 of 184 +/- 30 nmol/l (n = 6-10) but was much less effective in cardiac and smooth muscle (IC50s of 19.5 +/- 5.4 micromol/l (n = 6-12) and 37.9 +/- 1.0 micromol/l (n = 5-10), respectively). In all three tissues, the action of the drug on whole-cell KATP currents was rapidly reversible. In inside-out patches on beta-cells, gliclazide (1 micromol/l) produced a maximum of 66 +/- 13 % inhibition (n = 5), compared with more than 98 % block in the whole-cell configuration. CONCLUSION/INTERPRETATION: Gliclazide is a high-potency sulphonylurea which shows specificity for the pancreatic beta-cell KATP channel over heart and smooth muscle. In this respect, it differs from glibenclamide. The difference in the maximal block observed in the excised patch and whole-cell recordings from beta-cells, may be due to the absence of intracellular Mg nucleotides in the excised patch experiments. PMID- 11484081 TI - Increased islet DNA synthesis and glucose-derived lipid and amino acid production in association with beta-cell hyperproliferation in normoglycaemic 60 % pancreatectomy rats. AB - AIMS/HYPOTHESIS: Glycaemia does not change following a 60 % pancreatectomy in rats because of enhanced beta-cell function and proliferation (so-called beta cell adaptation). We previously studied these rats 4 weeks after surgery and showed hypersensitization of glucose-induced insulin secretion because of increased glucokinase activity. In this study of 60 % pancreatectomy rats 5 days after surgery, when beta-cell proliferation increased threefold, we investigated whether increases in glucose metabolism enhance the production of glucose-derived lipid, amino acids and DNA. METHODS: Isolated islets from 60 % pancreatectomy and sham-operated control rats 5 days or 4 weeks after surgery were studied. RESULTS: Five days after 60 % pancreatectomy surgery, islet glucose phosphorylation increased threefold, but overall glucose usage increased only twofold. The glucose-6-phosphate (G6P) concentration thus doubled, resulting in a sixfold increase in G6P metabolism through the pentose phosphate shunt (PPS). The pentose phosphate shunt generates ribose-5-phosphate for nucleotide synthesis, and DNA synthesis doubled in the partial pancreatectomy islets. In contrast, partial pancreatectomy rats 4 weeks after surgery had a smaller increase in glucokinase activity and their islet glucose-6-phosphate concentration and pentose phosphate shunt activity were equal to that of the control rats. DNA synthesis and beta cell proliferation, based on BrdU incorporation were close to normal. Another consequence of the heightened glucose metabolism in the 5-day partial pancreatectomy islets was twofold increase in production of glucose-derived lipid and the amino acids, alanine and glutamate. CONCLUSIONS/INTERPRETATION: The enhanced glucokinase activity in 60 % pancreatectomy rats supports the compensatory beta-cell hyperproliferation by increasing production of glucose derived DNA, lipids and amino acids. PMID- 11484082 TI - Inhibition of neointimal formation after balloon injury by cilostazol, accompanied by improvement of endothelial dysfunction and induction of hepatocyte growth factor in rat diabetes model. AB - AIMS/HYPOTHESIS: Cilostazol, a well-known phosphodiesterase type 3 (PDE3) inhibitor for the treatment of peripheral arterial disease, has vasodilator properties and an anti-proliferative action on the growth of vascular smooth muscle cells. In this study, we tested whether cilostazol inhibits neointimal formation and improves endothelial dysfunction after balloon injury in non diabetic and diabetic rats. METHODS: Cilostazol or vehicle was administered to non-diabetic and streptozotocin-induced diabetic rats from 7 days before to 14 days after balloon injury of the carotid artery. We focused on the expression of hepatocyte growth factor to explore how cilostazol improved endothelial dysfunction. Also, we studied the effects of cilostazol on hepatocyte growth factor production in in vitro experiments. RESULTS: At 14 days after injury, the ratio of neointimal to medial area was decreased in rats treated with cilostazol in non-diabetic and diabetic animals. The impaired response to acetylcholine in balloon injured vessels was improved by cilostazol in non-diabetic and diabetic rats (p < 0.05). Vascular hepatocyte growth factor concentration was decreased in injured vessels of non-diabetic rats compared to uninjured vessels. Moreover, hepatocyte growth factor was further decreased in injured vessels of diabetic rats as compared to those of non-diabetic rats (p < 0.05). Of note, administration of cilostazol attenuated the decrease in hepatocyte growth factor concentration in injured vessels of both non-diabetic and diabetic rats (p < 0.01). Increase in vascular hepatocyte growth factor by cilostazol was confirmed by in vitro experiments showing that cilostazol increased hepatocyte growth factor concentration in cultured human vascular smooth muscle cells, accompanied by cAMP accumulation. CONCLUSION/INTERPRETATION: Our study shows that the increase in vascular hepatocyte growth factor by cilostazol could improve abnormal growth of vascular smooth muscle cells and endothelial dysfunction through rapid regeneration of endothelial cells. PMID- 11484083 TI - Constitutive nitric oxide synthase is associated with retinal vascular permeability in early diabetic rats. AB - AIMS/HYPOTHESIS: We investigated the association between vascular permeability and constitutive nitric oxide synthase in rats with diabetes for a short duration (2 weeks). METHODS: Retinal vascular permeability was evaluated in rats with diabetes induced by streptozotocin using vitreous fluorophotometry and a small animal adapter. We carried out in situ hybridization and semi-quantitative reverse transcription-polymerase chain reaction to study the expression of endogenous constitutive nitric oxide synthase mRNA in diabetic retinas. We also examined changes in the protein expression of constitutive nitric oxide synthase in diabetic retinas using immunohistochemistry and Western blotting. RESULTS: Retinal vascular permeability was significantly higher in diabetic rats (median, 1.09 arbitrary unit) compared with control rats (median, 0.69 arbitrary unit) (p < 0.05). The expression of both neuronal nitric oxide synthase (NOS) and endothelial nitric oxide synthase mRNA was higher in diabetic retinas than in the retinas of control rats as determined by in situ hybridization and reverse transcription-polymerase chain reaction. Immunohistochemistry and Western blotting also showed that neuronal nitric oxide synthase increased in diabetic retinas. The immunohistochemistry of endothelial nitric oxide synthase indicated that non-vessel tissues increased in diabetic retinas while retinal vessels weakened. Western blotting showed that the amount of endothelial nitric oxide synthase increased. CONCLUSION/INTERPRETATION: These results suggest that increases in both constitutive NOSs (nNOS and eNOS) could be associated with retinal vascular permeability and that NOS is associated with clinical vascular dysfunction in the early stages of diabetes. PMID- 11484084 TI - Prevalence of coeliac disease in siblings of patients with Type I diabetes is related to the prevalence of DQB1*02 allele. AB - AIMS/HYPOTHESIS: Coeliac disease is more prevalent among patients with Type I (insulin-dependent) diabetes mellitus and coeliac disease-related antibodies have been reported to increase in frequency in their first-degree relatives. Our aim was to find out if coeliac disease is more common among siblings of children with Type I diabetes than in the normal population. METHODS: IgA endomysium antibodies were measured by indirect immunofluorescence in 550 subjects (mean age 11.8 years, range 3.1-26.9 years) with a sibling with Type I diabetes. We performed jejunal biopsy on as many subjects with positive antibodies as agreed. HLA-DQB1 genotyping was done in 427 subjects. RESULTS: Endomysium antibodies were positive in nine subjects (1.6 %). Jejunal biopsy was diagnostic for coeliac disease in five out of seven patients. An additional patient with coeliac disease, one already on a gluten-free diet, was identified by questionnaire. The prevalence of coeliac disease was 1.1 %. Five of six patients with coeliac disease had HLA DQB1*02 allele, compared with 118 of 421 of those without coeliac disease (p = 0.009). The sixth patient was positive for HLA-DQB1*0302 allele, which was also found in 241 of 421 of those without coeliac disease (p = 0.4). CONCLUSION/INTERPRETATION: We found the prevalence of coeliac disease among siblings of children with Type I diabetes to be similar to figures reported from recent population-based studies and to be correlated with the prevalence of coeliac disease associated HLA-DQB1 alleles. We propose that routine screening for coeliac disease among all first-degree relatives of patients with Type I diabetes is not warranted. PMID- 11484085 TI - Low rate of proliferation in immature thymocytes of the non-obese diabetic mouse maps to the Idd6 diabetes susceptibility region. AB - AIMS/HYPOTHESIS: The non-obese diabetic (NOD) mouse spontaneously develops T-cell dependent autoimmune diabetes. This mouse strain has a number of immune dysfunctions related to T-cell development but so far there are no available data on the proliferation of NOD immature thymocytes. We therefore studied the thymocyte proliferation in the NOD mouse in discrete stages of T-cell development. METHODS: We depleted thymocytes in vivo and analysed thymocyte proliferation during the thymus recovery from depletion. We used co-segregation analysis and quantitative loci trait analysis to investigate the genetic control of proliferation impairments in NOD thymocytes. RESULTS: Immature thymocytes of female NOD mice proliferate with a relatively low rate compared to non-autoimmune C57Bl/6 mice. This aberrant proliferation was most pronounced in CD4-/lo CD8+ cells differentiating from the CD4-CD8- to the CD4+CD8+ stage. A genetic mapping study using an F2 intercross between the NOD and the C57BL/6 strains showed that a major locus controlling this trait is linked to the insulin-dependent diabetes susceptibility locus Idd6. CONCLUSION/INTERPRETATION: Our results suggest that impairment of proliferation of immature thymocytes is one possible mechanism through which the Idd6 locus contributes to the pathogenesis of diabetes. PMID- 11484086 TI - Comment--to: Pinizzotto M, Castillo E, Fiaux M, Temler E, Gaillard RC, Ruiz J (2001) Paraoxonase 2 polymorphisms are associated with diabetic nephropathy in Type II diabetes. Diabetologia 44: 104-107. PMID- 11484088 TI - A novel mutation in islet amyloid polypeptide (IAPP) gene promoter is associated with Type II diabetes mellitus. PMID- 11484089 TI - The uncoupling protein 3 -55 C -->T variant is not associated with Type II diabetes mellitus in Danish subjects. PMID- 11484090 TI - Human arterial smooth muscle cells rapidly deplete cell culture media of glucose. PMID- 11484091 TI - Increased hospitalization with longer distance from treatment centre in diabetic paediatric patients in Germany. PMID- 11484092 TI - New challenges for orthopaedic advances in the twenty-first century. PMID- 11484093 TI - The role of Gd-enhanced three-dimensional MRI fast low-angle shot (FLASH) in the evaluation of symptomatic lumbosacral nerve roots. AB - In the field of lumbar spine disorders, three-dimensional (3-D) magnetic resonance imaging (MRI) can clearly depict a lumbar nerve root from the distal region to the dorsal root ganglion. In this study, we used a gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) enhanced-three-dimensional (3-D) fast low-angle shot (FLASH) sequence when examining lumbosacral disorders. The subjects were 33 patients (14 men and 19 women) in whom lumbosacral neural compression had been diagnosed clinically. Twenty-one patients had lumbar disc herniation, 11 had lumbar spinal stenosis, and 1 had lumbar radiculopathy caused by rheumatoid arthritis. Five subjects with low back pain were also studied as a control group. In all patients and in all 5 of the controls, the dorsal root ganglion of every root was enhanced clearly. There was no root enhancement in the 5 controls. Enhancement of the symptomatic nerve roots, caused by compression, was found in 11 of the 33 patients. All 11 patients had radiculopathy, and muscle weakness was more frequent in patients with enhanced nerve roots than in those without enhancement. There was no enhancement of the cauda equina, even in the patients with cauda syndrome. The enhancement effect may reflect some pathological condition of the compressed nerve root and needs to be studied further. PMID- 11484094 TI - Evaluation of the longitudinal arch of the foot with hallux valgus using a newly developed two-dimensional coordinate system. AB - The purposes of the present study were to establish a method for the evaluation of the longitudinal arch, using a two-dimensional coordinate system, and to clarify the main point of flattening of the foot in patients with hallux valgus. Measurements on a weight-bearing lateral radiograph of the foot were performed in 186 feet of 110 women with symptomatic hallux valgus and in 93 normal feet of 72 women. A two-dimensional coordinate system was used to perform the measurements. In this coordinate system, the line connecting the lowest point of the first metatarsal head and the lowest point (O) of the calcaneus is fixed as the x-axis, and the line that is perpendicular to the x-axis and intersects the x-axis at point O is fixed as the y-axis. The mean values of the y-coordinates at the points of the medial longitudinal arch of the foot in the patients with hallux valgus were significantly lower than those in the normal subjects (P < 0.001). The values were noticeably lower at points on the navicular. However, no change was observed in the posterior talus body. The new coordinate system indicated that the main points of flattening of the foot in patients with hallux valgus were the plantar flexion of the talus and the sinking of the navicular. PMID- 11484095 TI - Fibrin monomer could be a useful predictor of pulmonary embolism after total hip arthroplasty: preliminary report. AB - We examined 17 total hip arthroplasty patients in order to develop a method for the predictive diagnosis of pulmonary embolism (PE) after joint arthroplasty. Scintigraphy revealed the presence of PE in 4 patients. Prothrombin time (PT), activated partial thromboplastin time (aPTT), antithrombin III (ATIII), and thrombin-AT III complex (TAT) did not show significant differences between patients with and without PE. D-dimer 7 days after surgery showed significant differences between patients with and without PE. Fibrin monomer (FM) increased sharply after surgery, and it was significantly different between the patients with and without PE immediately after surgery and 2 days after surgery. Our findings suggest the importance of FM in the predictive diagnosis of pulmonary embolism after total hip arthroplasty, and 40 microg/ml or higher levels with our measurement method could represent a high-risk condition. PMID- 11484096 TI - Selective measurement of digital nerve conduction velocity. AB - We developed a new method to measure the nerve conduction velocity of a single digital nerve. In 27 volunteers (27 hands), we separately stimulated each digital nerve on the radial and ulnar sides of the middle and ring fingers. A double peaked potential was recorded above the median nerve at the wrist joint when either the radial-side nerve or the ulnar-side nerve of the middle finger was stimulated. The first peak of this potential had disappeared after the digital nerve was blocked under the stimulating electrodes, and the peak appeared again coinciding with the decrease of anesthesia. Shifting the stimulating electrodes on the digital nerve resulted in no significant difference in the peak conduction velocity. It is possible that each peak of the potential was attributable to conduction of an action potential along one of the two digital nerves. This new method allows the assessment of a single digital nerve, and may be clinically useful for assessing the rupture of a digital nerve and the sensory nerve action potentials in carpal tunnel syndrome. PMID- 11484097 TI - Effect of exercise training and detraining on bone mineral density in postmenopausal women with osteoporosis. AB - We examined the effect of exercise training and detraining on bone mineral density (BMD) in postmenopausal women with osteoporosis. Thirty-five postmenopausal women with osteoporosis, aged 53-77 years, were randomly assigned to three groups: a control group (n = 20), a 2-year exercise training group (n = 8), and an 1-year exercise training plus 1-year detraining group (n = 7). Exercise training consisted of daily brisk walking and gymnastic training. Calcium lactate, 2.0 g, and 1alpha-hydroxyvitamin D3, 1 microg were supplied daily to all subjects. No significant differences in initial lumbar BMD, measured by dual-energy X-ray absorptiometry (DXA) were found among the three groups. The mean percent change in BMD compared with the baseline was significantly higher at 1 and 2 years in the exercise training group and at 1 year in the detraining group than in the control group, and did not differ significantly at 2 years between the detraining and control groups. These findings indicate that our exercise training program led to a significant increase in lumbar BMD in postmenopausal women with osteoporosis compared with the control, but that the BMD reverted toward a level that was not significantly different from the control with detraining. Continued exercise training is needed to maintain the bone mass gained through exercise training. PMID- 11484098 TI - Effect of intermittent cyclical treatment with etidronate disodium (HEBP) and calcium plus alphacalcidol in postmenopausal osteoporosis. AB - We evaluated intermittent cyclical treatment with etidronate disodium (HEBP) and calcium plus alphacalcidol in postmenopausal osteoporosis, with special reference to bone mineral density (BMD) and prevention of spinal fracture. The patients were 40 women, over 50 years of age, with lumbo-dorsal pain and low BMD (less than 0.70 g/cm(2)), measured by dual-energy X-ray absorptiometry (DXA). The patients were randomly assigned to two groups. The first group (HEBP) received 200 mg of HEBP per day for 2 weeks, followed by 2 g calcium lactate and 0.5 microg alphacalcidol per day for the next 10 weeks. This 12-week cycle was repeated eight times for 2 years. The second group (Ca. D) received 2 g calcium lactate and 0.5 microg alphacalcidol per day for 2 years. Lumbar BMD was measured before the treatment and every 6 months during the treatment until 24 months, and changes were evaluated. The number of fractured vertebrae was counted on X-ray films before treatment and at the final assessment. After 6 months of treatment, a significant and continuous increase in BMD was observed in the HEBP group. Moreover, the percentage of patients with new vertebral compression fractures in the HEBP group was one-tenth of that in the Ca. D group. These results suggest that intermittent cyclical treatment with HEBP and calcium plus alphacalcidol may be effective for increasing BMD and preventing fractures in postmenopausal osteoporosis. PMID- 11484099 TI - Long-term results of rotational acetabular osteotomy in patients with slight narrowing of the joint space on preoperative radiographic findings. AB - Between 1975 and 1984, we performed rotational acetabular osteotomy in 22 female patients with painful hip dysplasia. At the time of surgery, the patients were in their twenties, and radiographs showed slight narrowing of the joint space. Of these patients, 15 were followed-up for 15 to 22 years (average, 19.8 years) after surgery. The preoperative severity of coxarthrosis in all 15 hips was graded as stage II, according to the classification of coxarthrosis advocated by the Japanese Orthopaedic Association. All 15 patients available for follow-up had had no additional operations on the operated side during the follow-up period. At the time of follow-up, the patients were aged 41 to 48 years (average, 44.3 years). Of the 15 patients, 12 had little or no pain and 14 could walk for more than 30 min without a cane; the severity of coxarthrosis was graded stage I in 3 hips, stage II in 4 hips, stage III in 5 hips, and stage IV in 3 hips. We conclude that rotational acetabular osteotomy is efficacious for patients who have preoperative radiographic findings of slight narrowing of the joint space. PMID- 11484100 TI - Flexible hinge toe implant arthroplasty for rheumatoid arthritis of the first metatarsophalangeal joint: long-term results. AB - We report the long-term clinical results and survival rate of the implant in flexible hinge toe implant arthroplasty of the first metatarsophalangeal joint, combined with a shortening oblique osteotomy of the metatarsal neck in the lateral toes, in patients with rheumatoid arthritis. Between 1983 and 1990, arthroplasty was performed on 97 feet in 66 patients. Twenty-seven patients died; follow-up information was available for 60 feet in the remaining 39 patients, who were followed for an average of 12 years. Twenty-nine patients (74%) were satisfied with the outcome after surgery, 7 were satisfied but had some pain or recurrent deformities, and 3 were unsatisfied. Radiologically, visible fracture was identified in nine implants. Four implants were removed because of infection (n = 2) or recurrent deformity (n = 2); no implant was removed because silicone synovitis developed. With revision as the endpoint, the implant survival rate was 93% at 10 years, and with radiographic implant fracture as the endpoint, the implant survival rate was 87% at 10 years. Shortening oblique osteotomy of the lateral toes appeared to decrease the rate of implant fracture and should be performed concomitantly with implantation when rheumatoid forefoot deformities are being reconstructed. PMID- 11484101 TI - The effect of energy restriction, walking, and exercise on lower extremity lean body mass in obese women with osteoarthritis of the knee. AB - Two hundred and twenty-eight Japanese women, aged 45-69 years, with osteoarthritis of the knee (knee OA) and a body mass index of greater than 26.4 self-selected one of the following interventions: (1) a nonsteroidal anti inflammatory drug (NSAID) alone (NA group; n = 52), (2) the NSAID with nonweight bearing exercises (NE; n = 49), (3) the NSAID combined with walking (NW; n = 35), (4) an energy restriction diet plus the NSAID (ND; n = 29), (5) the diet combined with the NSAID and strengthening exercises (NDE; n = 37), or (6) the diet combined with the NSAID and walking (NDW; n = 26). Body weight and lower extremity lean body mass (L-LBM), assessed with segmental bioelectrical impedance, were measured at the outset and at 8 weeks in the six therapy groups. Interval changes in L-LBM and L-LBM per body weight (L-LBM/weight) were compared among the six groups at the conclusion. The Lequesne index can index of severity of osteoarthritis of the knee at the final assessment, compared with that at the initial assessment, was significantly decreased in the NE group, but not in the NW or the NA groups (P = 0.023). There was a significant increase in the L LBM/weight in the NE group, but not in the NA or NW groups (P = 0.002 between NE and NA; P = 0.019 between NE and NW). There was a significant reduction in the Lequesne index in the NDE group in comparison with the ND and NDW groups (P = 0.0001 between NDE and ND; P = 0.0001 between NDE and NDW). There was a significant increase in L-LBM/weight in the NDE group, but not in the NDW or the ND groups (P = 0.011). Adjunctive therapy with nonweight-bearing lower extremity exercises appears to be more efficacious for prevention of lower extremity loss of muscle mass than adjunctive therapy with walking for obese women with knee OA. PMID- 11484102 TI - Is the proliferation of human chondrocytes regulated by ionic channels? AB - As in cells in all living cell systems, human chondrocytes are provided with a membrane potential. The existence of ion channels in the cell membrane is an essential prerequisite for the development of membrane potential. In nonhuman chondrocytes, different ion channels have already been identified. An association between potassium channel activity and cell proliferation has been detected in different human cell systems, whereas proof of an association between ion channel activity in human chondrocytes and their proliferation has yet to be established. In this study, we investigated the concentration-dependent influence of the ion channel modulators tetraethylammonium (TEA), 4-aminopyridine (4-AP), 4',4'diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS), 4-acetamido-4' isothiocyano-2,2'-disulfonic acid (SITS), and verapamil (vp) on the membrane potential and the proliferation of human chondrocytes, using flow cytometry. We found that the used ion channel modulators caused a change in the membrane potential of human chondrocytes. The membrane potential was decreased by 18% with 0.25 mmol/l vp (the maximal measurable effect of any of the ion channel modulators) compared with that in a control group. We measured DNA distribution in the human chondrocytes, and it was apparent that they were diploid cells with a very low proliferative tendency. These results allow us to conclude that ion channel modulators influence chondrocyte proliferation. Knowledge of the regulation of chondrocyte proliferation via ion channel modulators could serve in the research of new osteoarthritis treatment concepts. PMID- 11484103 TI - Comparison of absorbable poly-L-lactide and metallic intramedullary rods in the fixation of femoral shaft osteotomies: an experimental study in rabbits. AB - An osteotomy of the distal femoral diaphysis was fixed with an intramedullary self-reinforced poly-L-lactide (SR-PLLA) rod in 22 and with a metallic rod in 38 skeletally mature rabbits. Histomorphometric and quantitative computed tomography (QCT) were performed to assess time-related changes in the consolidation process of the osteotomy of cortical bone and development of the external callus. The follow-up times were 8, 16, 24, and 48 weeks for radiologic and 16, 24, and 48 weeks for histologic studies; 19/22 of the SR-PLLA and 34/38 of the metallic fixed osteotomies healed during the observation periods. On microscopic evaluation, no difference was found in histologic parameters such as external callus area and fraction, and endosteal callus area and fraction between SR-PLLA and metallic fixed osteotomies. The total external callus area increased up to 16 weeks and subsequently decreased linearly over time in both experimental groups. However, the mean endosteal bone area and fraction decreased after 16 weeks in metallic fixated femora, while in the SR-PLLA fixated femora, both these values increased, being greater in the SR-PLLA group. The mean external callus QCT density decreased after 8 weeks in both experimental groups, the decrease being greater in the SR-PLLA group. Nevertheless, no significant difference was detected between SR-PLLA and metallic fixed femora in any of the follow-ups. These results suggest that both SR-PLLA and metallic rods are suitable in the fixation of femoral shaft osteotomies in rabbits. Furthermore, SR-PLLA rod fixation seems to have a minor stress-shielding effect. PMID- 11484104 TI - Evaluation of long-term sequential changes in bone mass and strength following withdrawal of incadronate disodium (YM175) in ovariectomized rats. AB - We evaluated the long-term effects of withdrawal of a newer third-generation bisphosphonate, incadronate disodium (YM175), on both cancellous and cortical bone mass and strength in ovariectomized (OVX) rats. One hundred and sixty female SD rats at 13 weeks of age were randomized into four groups: sham-operated, OVX, and low- and high- YM (0.01 or 0.1 mg/kg s.c., three times a week after OVX). After 4 weeks of treatment with vehicle or incadronate disodium, rats from each group (n = 8) were killed at 0 (baseline), 3, 6, 9, and 12 months after the withdrawal of YM175. Histomorphometric studies of the proximal tibia revealed a dose-dependent decrease in OVX-induced bone turnover; cancellous bone volume was significantly higher in the YM groups compared with the OVX control group up to 6 months after withdrawal at low dose and up to 12 months after withdrawal at high dose. The low-dose group showed little effect on tibial diaphyseal cortical bone volume and width, while the high-dose group preserved both cortical parameters 12 months after withdrawal. Mechanical testing of femurs revealed that both metaphyseal and diaphyseal strengths were significantly higher at high dose compared with the OVX group until 12 months after withdrawal. These observations demonstrated that high-dose incadronate disodium preserved both cancellous and cortical bone mass and strength in OVX rats for 12 months after withdrawal of the agent. PMID- 11484105 TI - Effects of nicotine on the intervertebral disc: an experimental study in rabbits. AB - Clinically, we have observed that a large proportion of patients presenting with low back pain as the chief complaint are smokers. It was therefore postulated that smoking might affect the intervertebral disc. We investigated the histological and functional effects of nicotine on intervertebral discs in rabbits. Rabbits were implanted subcutaneously with minipumps for the delivery of 200 microg/ml nicotine for 4 or 8 weeks. The selected dose produced blood nicotine levels equivalent to those found in heavy smokers (30 cigarettes/day). Nicotine injection caused necrosis and fibrous tissue and vitreous formation in the nucleus pulposus of the intervertebral disc, as well as hypertrophy of the fibrous ring, with partial cracks and detachment. Measurement of collagen and proteoglycan production in intervertebral discs showed reduced synthesis of these proteins in nicotine-treated rabbits compared with the control findings. Our results indicated that the harmful effects of nicotine on the integrity of the intervertebral disc might be mediated by the direct effect of nicotine, or indirectly, by causing vasoconstriction of the vascular network surrounding the intervertebral discs. PMID- 11484106 TI - Involuted intraosseous lipoma of the sacrum showing high signal intensity on T1 weighted magnetic resonance imaging (MRI). AB - Involuted intraosseous lipoma with extensive fat necrosis resulting in cyst formation (Milgram stage III) is distinguishable from lesions without necrosis (stage I) or lesions with focal fat necrosis (stage II), based on differences in signal intensity on magnetic resonance imaging (MRI). Fat tissue has a high signal intensity on both T1- and T2-weighted MR images, whereas the extensive fat necrosis that results in cyst formation shows high signal intensity on T2 weighted images and low intensity on T1-weighted images. We report a patient in whom an intraosseous lipoma with high signal intensity on both T1- and T2 weighted MRI was found to be extensively involuted on histopathologic examination. Intraosseous lipoma appears to undergo spontaneous involution. In some patients, therefore, surgical excision may not be necessary. A correct preoperative diagnosis should reduce the necessity for a biopsy or surgery. Although lesions classified as stage I or II are easily identified by MRI, those of stage III are difficult to diagnose preoperatively by this method. PMID- 11484107 TI - Traumatic bilateral quadriceps tendon rupture. AB - Bilateral spontaneous quadriceps tendon rupture is a very rare event, with only an estimated 40 cases reported in the literature. We report a case of bilateral spontaneous quadriceps tendon rupture and review the literature. The reviewed literature recommends early repair; therefore, early diagnosis is crucial. Reportedly, up to 50% of spontaneous bilateral quadriceps ruptures are misdiagnosed at first, resulting in poorer outcome. As spontaneous bilateral quadriceps rupture has previously been reported in association with chronic disease, with, to date, one-third of cases reported to show this association, routine laboratory screening for rheumatologic, endocrine, and renal disease is recommended for all presenting patients. We also briefly discuss the need for antithrombotic prophylaxis, as our patient died, suddenly with the clinical symptoms of a pulmonary embolism, 7 days after surgery and immobilization. PMID- 11484108 TI - Multiple intramuscular metastases 15 years after radical nephrectomy in a patient with stage IV renal cell carcinoma. AB - We report an 81-year-old man with late recurrent multiple metastases in skeletal muscles 15 years after radical nephrectomy was carried out for stage IV renal cell carcinoma (RCC). The tumors were located in the left triceps muscle and the brachioradial muscle. We performed surgical resections of both tumors, and histological analysis revealed that they were both metastatic RCC of the clear cell type. In this case report, we discuss the characteristics and differential diagnosis of this tumor on magnetic resonance imaging, and we also refer to the rarity of muscle metastasis and the unpredictable tendency of this tumor to remain dormant for long periods. PMID- 11484109 TI - The association of neurofibromatosis 1 and spinal deformity with primary hyperparathyroidism and osteomalacia: might melatonin have a role? AB - A 35-year-old woman with neurofibromatosis 1 and thoracic kyphoscoliosis had incomplete paraplegia. She had a history of hyperparathyroidism due to a parathyroid adenoma which had been excised 4 years previously. Plain radiographs of the spine revealed kyphoscoliosis from the third to sixth thoracic vertebrae. Kyphosis and scoliosis angles were 86 degrees and 28 degrees, respectively. Radiographs of the skull and hands showed radiological changes suggestive of hyperparathyroidism. Laboratory tests showed low-normal serum calcium, hypophosphatemia, elevated serum alkaline phosphatase, and low serum 25 hydroxyvitamin D. Retrospective review of the patient's laboratory data showed that she had osteomalacia at the time of diagnosis of primary hyperparathyroidism. The patient had been treated by anterior and posterior decompression and fusion with posterior instrumentation through a single posterior approach. The postoperative kyphosis and scoliosis angles were 30 degrees and 12 degrees, respectively. Neurological recovery and spinal fusion had been achieved. Osteomalacia responded well to vitamin D therapy. This is the first case of coexisting neurofibromatosis 1, primary hyperparathyroidism due to parathyroid adenoma and osteomalacia to be reported in the literature. The osteomalacia in this patient could be related to primary hyperparathyroidism, and not to neurofibromatosis 1. A drop in melatonin level after parathyroidectomy may have been the cause of spinal curvature progression in this patient. PMID- 11484110 TI - Orthopedic applications of gene therapy. AB - A literature review of the applications of gene therapy for the treatment of orthopedic disorders was conducted, and showed that gene therapy provides us with new possibilities for the clinical management of orthopedic disorders. Most of these disorders, such as failure to obtain spinal fusion, disc degeneration, fracture and segmental bone defects, bone tumor, articular disorders, soft-tissue injury, genetic disorders, and nerve and muscular disorders, are difficult to treat by traditional surgical or medical means, and are under investigation for gene therapy. Many rapid advances have been made in the field of this novel approach. Although a number of obstacles remain to be solved before gene therapy can be applied for clinical use in humans, it is already apparent that gene therapy has the great potential of becoming a valuable clinical treatment for orthopedic disorders in the twenty-first century. With the development of gene transfer techniques, gene therapy will probably have numerous applications in orthopedic disorders. PMID- 11484111 TI - Research and publication in orthopaedics. PMID- 11484112 TI - Bone and soft-tissue tumors in postgraduate education. PMID- 11484113 TI - Morphological characteristics of the bony birth canal in patients with developmental dysplasia of the hip (DDH): investigation by three-dimensional CT. AB - We investigated the three-dimensional morphological characteristics of the pelvis in adult female patients with developmental dysplasia of the hip (DDH), using computerized tomography (CT) images. Forty-two subjects with normal hips and 40 DDH patients were recruited for the study. In the DDH group, the average transverse diameter of the pelvic inlet was significantly less and the average transverse diameter of the pelvic outlet was significantly greater than the measurements in the normal group. Further, the bony birth canal in DDH patients exhibited a higher incidence of anthropoid-type geometry, as defined by a longer sagittal diameter relative to the transverse diameter of the pelvic inlet. These findings indicate a characteristic pelvic geometry and suggest different development of the pelvis in the transverse direction in DDH patients. In addition, the obstetric conjugate length/transverse diameter of the pelvic inlet ratio was correlated to the degree of severity of acetabular dysplasia. This finding suggests that DDH is a manifestation of a developmental characteristic of the pelvis. PMID- 11484114 TI - Relationship between cervical curvature index (Ishihara) and cervical spine angle (C2--7). AB - The purpose of this study was to determine the relationship between the cervical curvature index (Ishihara) and the cervical spine angle (C2--7) from both cervical lateral radiographs and a geometrical model. The cervical lateral radiographs of 295 outpatients with no cervical symptoms were reviewed to measure the cervical curvature index (Ishihara) and the cervical spine angle (C2--7). The average cervical curvature index (Ishihara) was 10.9 (SD, 15.3) and the average cervical spine angle (C2--7) was 20.3 (SD, 14.3). A highly significant correlation (0.95; 95% confidence interval [CI], 0.94--0.96) was found between the cervical curvature index (Ishihara) and the cervical spine angle (C2--7). Their linear correlation was also validated in the geometrical model. Their correlation diminished, however, in subjects with an S-shaped cervical spine. The cervical curvature index (Ishihara) and the cervical spine angle (C2--7) were interchangeable, but this interchangeability was lost in the S-shaped cervical spines. PMID- 11484115 TI - Relationships between physical activity and metacarpal cortical bone mass and bone resorption in hemiplegic patients. AB - The aim of the present study was to investigate the relationships between physical activity and metacarpal cortical bone mineral density (BMD) and bone resorption in hemiplegic patients. Seventy-two male hemiplegic patients with cerebrovascular disease (mean age, 67.0 years; range 48-83 years) were recruited. Metacarpal cortical BMD in bilateral hands was measured by computed X-ray densitometry, and the levels of urinary cross-linked N-telopeptides of type I collagen (NTx), as a bone resorption marker, were measured by an enzyme-linked immunosorbent assay (ELISA). BMD (mean +/- SD) in the paralyzed hand was significantly lower than that in the nonparalyzed hand (2.52 +/- 0.46 and 2.64 +/ 0.45 mmAl; P < 0.05). On multiple regression analysis, BMD was significantly positively correlated with Brunnstrom stage (assessment of degree of paralysis) in the hand (P < 0.05), whereas urinary NTx levels were significantly negatively correlated with Brunnstrom stage in the hand, arm, and leg, and with walking ability (P < 0.05, P < 0.05, P < 0.01, and P < 0.01, respectively). These findings suggest that, in male hemiplegic patients, although metacarpal cortical BMD may be affected by only local physical activity, urinary NTx appears to be responsive to local and general physical activity. PMID- 11484116 TI - Acute nonspecific flexor tenosynovitis in the digits. AB - Eight patients, aged 53 to 86 years (average, 63 years), with typical symptoms of acute infectious digital flexor tenosynovitis after excessive use of the hand were treated. No bacterial or crystal association was proven. In the two patients treated earliest, antibiotics were administered because of leucocytosis and a positive C-reactive protein (CRP) test result, but there was no improvement with this treatment. Surgical treatment relieved the symptoms in both these patients. In the other six patients, hand elevation and rest, as well as treatment with anti-inflammatory agents, resulted in complete resolution of the symptoms. Considering that the structure of the digital flexor tendon and the tendon sheath is similar to that of the synovial joint, the author suggests that the nonspecific tenosynovitis in this series could have been caused by overload on a flexor tendon system that had degenerated with age. PMID- 11484117 TI - Treatment of humeral shaft nonunion by external fixation: a valuable option. AB - We report the treatment of six patients with nonunion of the humerus, using a unilateral fixator and bone grafting. Union was obtained in all patients, with an average time to union of 4.5 months. Superficial pin tract infection was seen in five patients, but resolved uneventfully. One patient had transient radial nerve palsy. The results, according to the Stewart and Hundley criteria, were excellent in one patient, good in three, fair in one, and poor in one. The main cause of the fair and poor results was marked limitation of shoulder and elbow motion. This method, however, seems to be therapeutically effective. PMID- 11484118 TI - Effect of administration of lipoprostaglandin E(1) on physical activity and bone resorption in patients with neurogenic intermittent claudication. AB - The effect of the systemic administration of prostaglandin (PG) E(1) on physical activity and bone resorption was examined in patients with intermittent claudication (gait disturbance). Twenty male patients (age, 67.2 +/- 7.8 years; mean +/- SD) with neurogenic intermittent claudication caused by lumbar spinal canal stenosis were included in the study. Lipo-PG E(1) was administered intravenously, at a daily dose of 10 microg, on 3 days a week, for 6 months in all patients. Clinical symptoms, levels of urinary cross-linked N-telopeptides of type I collagen (NTx), and metacarpal cortical bone mineral density (BMD) were assessed before and just after the 6 months of treatment. Subjective symptoms, including leg pain and/or tingling and gait disturbance, and restrictions of the activities of daily living were significantly improved. However, no significant changes were observed in either urinary NTx levels or metacarpal cortical BMD. These findings suggest that the systemic administration of PG E(1) appears to improve subjective symptoms and activities of daily living in elderly male patients with neurogenic intermittent claudication, but does not affect either bone resorption or metacarpal cortical BMD. Short-term systemic administration of PG E(1) and increased physical activity in elderly male patients with gait disturbance caused by lumbar spinal canal stenosis may not affect bone resorption. PMID- 11484119 TI - Factors affecting range of flexion after total knee arthroplasty. AB - Many factors affect postoperative range of flexion after total knee arthroplasty (TKA). The purpose of this study was to identify the most important factors that affect range of knee flexion after TKA. Sixty-five patients (73 knees) were treated with Genesis II knee replacements. Minimum follow-up was 2 years. Patient demographics (sex, age, body mass index, previous surgery, component type, patella resurfacing, preoperative Knee Society score preoperative range of motion) and radiographic measurements (preoperative tibiofemoral varus/valgus angle, height of the joint line, length of the patellar tendon, shift and tilt angle of the patella) were analyzed statistically. Among these factors, preoperative range of flexion, positively, and preoperative varus/valgus tibiofemoral angle, negatively, affected postoperative range of flexion. The tilt angle of the patella and the tilt angle of the patellar button approached statistical significance in revealing a negative relation with postoperative range of flexion. TKAs in which the patella was not resurfaced tended to lose range of flexion, whereas TKAs in which the patella was resurfaced tended to have no loss of flexion, although the number of TKAs in which the patella was not resurfaced was small. In conclusion, preoperative range of flexion and preoperative varus/valgus tibiofemoral angle affected postoperative range of flexion. The tilt angle of the patella and tilt angle of the patellar button may be factors that affect postoperative range of flexion. PMID- 11484120 TI - Effect of implant lengthening and mode of fixation on knee laxity after ACL reconstruction with an artificial ligament: a cadaveric study. AB - An "apparent" lengthening of the ligament implant, which causes an increase in knee laxity after the reconstruction of the anterior cruciate ligament (ACL) may be due to either slippage of the implant from under the fixation devices, or tunnel migration (due to bone resorption). These two mechanisms are related to the initial ligament placement, implant tensioning, and fixation modes. This cadaveric study simulates, in a controlled experimental situation, the postoperative lengthening of artificial ACL implants, and seeks to quantify the consequent increase in joint laxity. Eight cadaveric right knees, in which the Leeds-Keio artificial ligament was implanted, were tested in a specially constructed apparatus, which allowed the knee joint six degrees of freedom. In each of the tested joints the laxity was measured under several test conditions for two final fixation modes of the implant. The difference between the fixation modes was the application (as in mode B) or not (as in mode A), of a posteriorly directed force of 50 N on the tibia, at the moment of final fixation of the ligament. In both cases a tensile load of 50 N was maintained along the implant. All measurements were taken at flexion angles of 20 degrees and 90 degrees and with controlled implant lengthening of up to 3 mm in 0.5-mm increments. After implantation, adopting fixation mode B resulted in the knee exhibiting an anterior laxity considerably less than the original physiological laxity, compared with that measured after using fixation mode A. Thus at 20 degrees of knee flexion, under an anterior load of 100 N applied on the tibia, adopting fixation mode B, the joint laxity was 2.8 mm smaller than the natural laxity, whereas, for fixation mode A, it was 1.4 mm larger. At 90 degrees of knee flexion, the situation was similar, but with smaller differences. However, the situation was overturned as the implant length was increased. Thus, at 20 degrees of knee flexion, when the implant was lengthened in a range of 1--2 mm, the laxity observed with fixation mode B was similar to that recorded when the ACL was intact, whereas the laxity observed with fixation mode A was about 3--4 mm greater. Similar data were observed at 90 degrees of knee flexion. It appears that fixing the implant finally by applying a tensile load on it while simultaneously pushing the tibia posteriorly could be an effective measure against the possible return of joint laxity. PMID- 11484121 TI - Experimental study on slow-speed elongation injury of the peripheral nerve: electrophysiological and histological changes. AB - Slow-speed elongation of the sciatic nerves was induced in 41 rabbits by lengthening the osteotomized femur by 2.1 mm/day with an external fixator. Time courses were monitored. About 2 weeks after elongation began, the amplitude of the compound nerve action potentials decreased, and delayed latency began. Histological observations showed that the wavy structure of the nerve fibers was lost during the stage at which reduction in the amplitude of action potentials occurred. In some nerve fibers, narrowing of the axons was observed. Further elongation caused delay in the percent latency. Histologically, collagen fibers were noted in the nerve fascicles, as well as narrowing in all the nerve fibers. When elongation ceased at the stage at which a reduction in the amplitude of action potential was noted, recovery from nerve damage followed. When elongation did not cease until the stage at which latency was delayed, however, recovery was not apparent, either histologically or electrophysiologically. The critical level of percent amplitude for recovery was at 82.4% of the original level (the critical rate for bone lengthening was 20.5%). PMID- 11484122 TI - A PCR-ELISA assay for the detection of disseminated osteosarcoma cells in a mouse metastatic model. AB - Circulating tumor cells in the blood play a central role in the metastatic process. There have been no reports describing the relationship between lung metastasis and circulating osteosarcoma cells. We developed a system with a polymerase chain reaction assay based on an enzyme-linked immunosorbent assay (PCR-ELISA) to detect circulating osteosarcoma cells in a mouse metastatic model. Osf2/Cbfa1, hereafter called Osf2, a member of the runt family of transcription factors, was used as a target gene. One splicing variant of Osf2 mRNA was identified and its expression was restricted to the bones and osteosarcomas. The amount of the splicing variant of Osf2 mRNA was significantly higher in the blood of mice with metastasis than in the blood of the control group. The PCR-ELISA using Osf2 mRNA is a potential method to detect circulating osteosarcoma cells in peripheral blood. PMID- 11484123 TI - Time-sequential changes in biomechanical and morphological properties of articular cartilage in cryopreserved osteochondral allografting. AB - This study examined time-sequential changes in the biomechanical and morphological properties of articular cartilage that had received cryopreserved osteochondral allografting. Osteochondral blocks obtained from the femurs of 18 rabbits were cryopreserved with dimethylsulfoxide (DMSO), using a two-step freezing method, and allografted to the femurs of another 18 rabbits. Specimens for biomechanical and morphological examinations were prepared at the second, fourth, and twelfth weeks after allografting (n = 18). In 12 allografted rabbits, biomechanical features were examined with an indentation test apparatus, and histological changes were studied with a light microscope (second week, n = 4; fourth week, n = 4; twelfth week, n = 4). In the other 6 allografted rabbits, cartilage surfaces were studied with a scanning electron microscope (second week, n = 2; fourth week, n = 2; twelfth week, n = 2). For controls, fresh, DMSO treated, or DMSO-treated + cryopreserved specimens were examined biomechanically and morphologically. In the time-sequential examination of biomechanical features, both the parameter for elasticity (i.e., ratio of instant elastic strain to maximum strain) and the parameter for viscosity (i.e., average retardation time) significantly changed. Light microscopy showed chronological decreases in safranin-O staining intensity in the matrix, and progression of degeneration. On scanning electron microscopy, disruption of the cartilage surface was also recognized. Therefore, changes in biomechanical properties due to cryopreservation could cause irreversible changes in the cartilage in cryopreserved osteochondral allografting. PMID- 11484124 TI - Severe pelvic pain and extrinsic compression of the rectum: late complication of total hip arthroplasty. AB - Various pelvic and visceral complications have been reported resulting from total hip arthroplasty. Most of these complications are known to be related to the intrapelvic migration of the acetabular cup or the cement, or to the heat generated by polymerization of the methylmethacrylate. Complications involving almost every pelvic structure have been described. We report a case of progressive limb shortening and severe pelvic pain beginning 6 months after total hip arthroplasty. To close the acetabular defect after removal of the displaced acetabular component, we used a modified extraperitoneal approach adapted from Stoppa's technique, which is commonly used for inguinal, femoral, and obturator hernias. In this article, we aim to describe this modified extraperitoneal approach and to offer advice for the diagnosis and treatment of this serious complication. PMID- 11484125 TI - Multiple osteochondroses of bilateral knee joints. AB - We experienced a patient with a combination of multiple osteochondroses: Blount's disease, bipartite patella, and Sinding-Larsen-Johansson disease in the left knee, and a combination of bipartite patella and Osgood-Schlatter disease in the right knee. The patient was a healthy, active 12-year-old boy with bilateral knee pain. He had been diagnosed with Blount's disease of the left tibia at 2 years of age, and had been treated with open wedge osteotomy. He was diagnosed with bilateral bipartite patellae at the age of 9 years, and was diagnosed with Osgood Schlatter disease in the right knee and Sinding-Larsen-Johansson disease in the left knee at 10 years of age. The second growth spurt was observed during this period. At 11 years of age, he was diagnosed with an osteochondral fracture of the left lateral femoral condyle and was observed without surgery. This patient showed the sequential appearance of an ossification disorder, probably due to the abnormal response of enchondral ossification to mechanical stress. Overuse in this growth period may have played a role in the development of these osteochondroses. The osteochondral fracture was probably caused by a disruption at one of the weakest parts of the developing skeleton, between the ossification center and the overlying cartilage in the background of an ossification disorder. PMID- 11484126 TI - The role of imaging modalities in the diagnosis of primary dedifferentiated parosteal osteosarcoma. AB - Dedifferentiated parosteal osteosarcoma (dd-POS) is defined as high-grade sarcomatous components coexisting with low-grade POS components. With regard to the histological diagnosis of dd-POS, the sampling of a small area of dedifferentiation through the densely mineralized POS can be a problem. In this situation, imaging is important to identify the area with the highest possibility of dedifferentiation. We report a patient in whom dedifferentiation was shown by computed tomography (CT) and magnetic resonance imaging (MRI). CT revealed a radiolucency in a highly mineralized area. T2-Weighted MRI showed a relatively high signal intensity, corresponding to the radiolucency, surrounded by a very low signal intensity area. Furthermore, gadolinium-enhanced T1-weighted MRI showed marked enhancement. Based on these imaging techniques, the condition was diagnosed as most likely to be a dd-POS, although a representative sample was not accessible by incisional biopsy. Neoadjuvant chemotherapy was administered, followed by wide resection and adjuvant chemotherapy. Four years after the surgery, partial lobulectomy was required because of a pulmonary metastasis. Three years after the second surgery, the patient remained well without evidence of metastases. Based on the initial diagnosis and, consequently, the optimal treatment of combined chemotherapy and wide resection, our patient showed a good clinical outcome. PMID- 11484127 TI - Primary adult human bone cells do not respond to tissue (continuum) level strains. AB - Bone adapts to its mechanical environment, and, since the late 1800s, investigators have presumed that this adaptation relates to strain magnitude. Indeed, overwhelming evidence supports the view that either strain or some strain related quantity stimulates bone adaptation or remodeling. Virtually all investigators, implicitly or explicitly, assume that the level of strain magnitude responsible for bone adaptation is that measured by strain gauges in vivo (i.e., 100-2500 microstrain) and that bone cells are directly deformed by strained matrix. We present evidence that bone cell deformation in this range does not cause bone adaptation. First, bone cells in vitro typically do not respond to average (continuum) levels of strain magnitude. Second, bone cells in vitro do respond to fluid flow-induced shear stresses in these ostensible physiological ranges. Third, in vivo strain magnitudes presumed to stimulate remodeling reflect only averages, and not local peaks, which are 2-15 times higher. Thus, we hypothesize that sensing cells do not respond to levels of strain presumed to be physiological. PMID- 11484128 TI - Revision of failed metal-backed patellar component of Miller/Galante-I total knee prosthesis. PMID- 11484129 TI - Feminist ethics and the metaphor of AIDS. AB - This paper looks at a range of metaphors used within HIV/AIDS discussions and research in support of the claim that bioethicists should pay serious attention to metaphors. Metaphors shape the ways we think about problems and the types of solutions we investigate. HIV/AIDS is an especially rich field for the investigation of metaphor, since the struggles for dominance among different metaphorical options has been very evident. In the field of medical research as well as in the area of public policy, different metaphors support different strategies and, therefore, the choice of metaphor has ethical significance. I argue that feminists should engage in these debates since they have an interest in the metaphors selected. They can also learn to be more self-conscious in choosing politically liberating metaphors in other bioethics contexts. PMID- 11484130 TI - Understanding autonomy relationally: toward a reconfiguration of bioethical principles. AB - Principle-based formulations of bioethical theory have recently come under increasing scrutiny, particularly insofar as they give prominence to personal autonomy. This essay critiques the dominant conceptualization of autonomy and urges an alternative formulation freed from the individualistic assumptions that pervade the prevailing framework. Drawing on feminist perspectives, I discuss the need for a vision of patient autonomy that joins relational experiences to individuality and acknowledges the influence of patterns of power and authority on the exercise of patient agency. Deficiencies in the current models of science and social relations guiding medical practice are analyzed, particularly (1) the tendency to disregard the patient's self-knowledge and (2) failure to recognize limitations on the generalizability of medical knowledge. Models of social relations such as mothering and friendship are explored to advance a conception of autonomy better suited to the practical activities of medicine. In conclusion, I consider how acknowledgement of the specificity and complexity of social relations can contribute to reconfiguration of other principles comprising the standard framework of bioethics, particularly beneficence, justice, and equality. PMID- 11484131 TI - Some speculations on matters of touch. AB - In this essay, I examine the question of whether it is possible that the encounter with the other could be mediated such that the interval of distance would lose its determining power. I reflect on some instances of extraordinary corporeality, most particularly the phenomenon of conjoined twins, in order to problematize the relation between subjects as they are embodied. Where the normative body is supposedly marked out by the closed boundaries of the skin, the figuration of the anomalous body as potentially leaky and contaminatory positions it as being of prime concern to both medical and cultural discourse. Within those paradigms there are of course many pressing bioethical issues to debate, but my intention is to draw back from a problem-based scenario in order to consider the threshold of ethics in the self/other relation itself. PMID- 11484132 TI - The concept of a feminist bioethics. AB - Feminist bioethics poses a challenge to bioethics by exposing the masculine marking of its supposedly generic human subject, as well as the fact that the tradition does not view women's rights as human rights. This essay traces the way in which this invisible gendering of the universal renders the other gender invisible and silent. It shows how this attenuation of the human in 'man' is a source of sickness, both cultural and individual. Finally, it suggests several ways in which images drawn from women's experience and women's bodies might contribute to a constructive rethinking of basic ethical concepts. PMID- 11484133 TI - Shifting the autonomy debate to theory as ideology. AB - Some feminists have been critical about the dominant conception of autonomy, questioning, for example, its conception of persons and ideal of personhood. Tom Beauchamp and James Childress (B&C), the major proponents of the dominant conception of autonomy, believe that these feminists have misunderstood their theory and, moreover, that their theory is immune to feminist attack. Their response to feminist critics, however, has been dismissive and does nothing to assuage these critics' concerns. In this paper I briefly review the state of play in this debate about autonomy, showing that B&C are not without positive rejoinders to objections raised by feminist critics. These rejoinders rest on the notion that feminist concerns are a matter of what is logically entailed by B&C's theory of autonomy and attempt to show that feminist commitments are logically consistent with that theory. However, these rejoinders are less than convincing for reasons illuminated by Cheshire Calhoun. Calhoun reminds us that feminists are sensitive to ways in which the shape of discourse is influenced by non epistemic considerations. In particular, Calhoun draws our attention to the cumulative effect of a whole tradition of moral reasoning that focuses on too narrow a range of moral problems and too narrow an understanding of people and the human condition. B&C's conception of autonomy relies on and reinforces ideologies of the moral life created in just this way. Following Calhoun, I show that criticism of their theory as ideology is not criticism of its logical implications, but something far more damaging, something without available rejoinders. PMID- 11484134 TI - Feminist bioethics and psychiatry. AB - Feminist bioethics is a relatively new field, the major works in which only started to appear in the late 1980s. At first feminist bioethicists focused mainly on issues of particular concern to women such as reproduction. Recently, papers have begun to appear that show that a feminist analysis can be brought to bear on any subject traditional bioethics discusses. So far, however, feminist bioethics has not been brought to bear on psychiatry. There have been feminist critiques of psychiatry and feminist discussions of certain diagnostic categories that disproportionately affect women, but these are concerned with women's issues within psychiatry and how psychiatry has been used to oppress women. Certainly these are important, but what has been missing is a discussion of psychiatry in the sense that feminist bioethics suggests a general critique of psychiatry and the rethinking of the practice of psychiatry, regardless of whether the specific instances involved are women's issues. In this paper I look at what such a feminist bioethical intervention into psychiatry would look like. PMID- 11484135 TI - [Polymeric nanoparticles: new systems for improving ocular bioavailability of drugs]. PMID- 11484136 TI - [Presbyopic surgery]. PMID- 11484137 TI - [Health transfers]. PMID- 11484138 TI - [Topical drugs absorption in ocular surface diseases]. PMID- 11484139 TI - [Ocular changes associated with connective tissue disorders: role of the elastic and collagen components]. AB - Every day, the emerging fields of genomics and proteomics provide new information about both normal and pathological processes, among which, the extracellular matrix appears to play a significant role. Over the past decade, the classic fibrillar components of the matrix, collagen and elastin, have been the subject of extensive research leading to a wealth of information which is far form being fully interpreted. Although the basic composition and structure of the matrix components has been well established, today more than twenty five different varieties of collagen have been described, and elastic fibers are currently described as polymeric complexes, composed of at least 19 different proteins in their microfibrillar and amorphic portions. Mutations in three of the genes coding for some of the most abundant proteins in the elastic fibers give rise to a wide range of elastic tissue phenotypes, from skeletal or dermal anomalies to vascular or ocular defects. In this review, our aim was to gain insight into the fibrillar component of the extracellular matrix in an attempt to improve our understanding of certain ocular disorders. PMID- 11484140 TI - [Trifluoperazine's action on fibroblast proliferation in cell cultures. Comparison of different doses]. AB - PURPOSE: To evaluate the effect of different doses of trifluoperazine (TFP) on fibroblastic proliferation in cell cultures of sclera and conjunctiva. METHODS: 24 brown rabbits were operated on for non-protected sclerectomy and were divided into four groups. Group 1: Subconjunctival injection of 0.5 ml of balanced saline solution (BSS) 24 h before surgery. Group 2: Intraoperative TFP 10(-2)M for five minutes on the sclera in experimental filtrating surgery. Group 3: Subconjunctival injection of 0.5 ml of TFP 10(-3) M 24 h before surgery. Group 4: Subconjunctival injection of 0.5 ml of TFP 10-4M 24 h before surgery. Conjunctiva and sclera tissue samples were taken 1 hour after surgery in the treated area and at 90 degrees from it. All of the samples were cultured in DMEM with 20% fetal calf serum. The presence of atypical cells and fibroblastic growth was assessed at 3, 6, 9 and 12 days. RESULTS: There were statistically significant differences in all the groups and in the times compared to the control group. In the treated conjunctiva, differences were found between the subconjunctiva 10(-3) M and the intraoperative 10(-2) M on the 3rd and 6th days, between the preoperative 10-2 and subconjunctiva 10(-3) M and 10(-4) after the 6th day. In conjunctiva at 90 degrees, there were significant differences between preoperative 10-2 M and subconjunctiva 10-3 after day 6, between the preoperative 10-2 M and subconjunctiva 10(-4) on days 9 and 12. There were no significant differences among the different treatments in the sclera samples, but there were differences between all them and the control group. CONCLUSIONS: TFP at a dose of 10(-2) and 10(-4) TFP has an inhibitory effect on cellular proliferation in vitro in the rabbit sclera and conjunctiva. All the doses used produce a significant inhibition of cellular proliferation without producing significant disorders in the cellular morphology. PMID- 11484141 TI - [Analysis of aged-related nerve fiber layer thickness changes with laser polarimetry]. AB - PURPOSE: Scanning laser polarimetry allows a quantitative study of the retinal nerve fiber layer thickness. It was used to evaluate the effects of aging on thickness of nerve fiber layer in the retinal of normal subjects. METHODS: The retinal nerve fiber layer thickness of 48 eyes of 48 normal subjects was studied. Three consecutive polarimetric images were acquired for each eye and the mean image was calculated, the nerve fiber layer thickness measurements was obtained in this image using the standard 10 pixels and 1.75 disc diameter band centered on the optic nerve head. The standard GDx quadrants were used. Linear regression analysis was used to evaluate the effects of aging on the nerve fiber layer thickness. RESULTS: The mean nerve fiber layer thickness was 78.59 SD 9.81 microm. A significant reduction with age was found in the mean nerve fiber layer thickness (r2: 0.19, p=0.002), the mean superior-inferior quadrant thickness (r2: 0.239, p<0.001), the superior (r2: 0.212; p=0.001) and inferior quadrant thickness (r2: 0.196, p= 0.002), and the maximal superior quadrant (r2: 0.111, p= 0.019) and inferior quadrant thickness (r2: 0.081, p=0,048). CONCLUSIONS: The nerve fiber layer thickness decreases with increasing age in normal subjects. PMID- 11484142 TI - [Predictive value of the pattern-electroretinogram in glaucoma]. AB - PURPOSE: To determine the ability of pattern-electroretinogram (PERG) to discriminate individual evolution of glaucomatous visual field losses in glaucoma suspects. METHODS: We studied 151 eyes of 78 GS, classified depending on their risk to develop glaucomatous damage. The initial visual field was normal. Those eyes showing at least 2 reduced amplitudes in the PERG components (P50, N95 and NP) were considered pathologic. The mean follow-up was 30 months (range: 13-50 months); the VF was evaluated at the end of the study. RESULTS: At the beginning of the study the PERG was pathologic in 65 eyes (43%); in 45 of these eyes (69%) the visual field was pathologic at the end of the study. The positive predictive value of the PERG of visual field glaucomatous changes was 69% and the negative predictive value was 80%. The N95 amplitude showed the best predictive value, with 100% and 77% in the high and medium risk groups, respectively. CONCLUSIONS: The PERG study can discriminate the glaucoma suspects that will develop visual field glaucomatous losses. PMID- 11484143 TI - [Epidemic optic neuropathy in Cuba. Clinical and neurophysiological study]. AB - The Epidemic Optic Neuropathy (EON) that occurred in Cuba in 1993 affected more than 50,000 people. With the purpose of characterizing it from the clinical and neurophysiological points of view, and of determining the etiology of this illness, we studied 85 of this patients who were treated in the Ophthalmology Department of the Hermanos Ameijeiras Clinical Surgical Hospital, located in Centro Havana, Havana city. Cuba. METHODS: We examined 8 cases at the beginning of the epidemic, 27 in the middle period and 50 at the end. RESULTS: The bilateral and symmetrical affectation of the optic nerve was the most remarkable clinical feature in all cases studied. This affectation was more severe in the first group of studied patients with a very extended evolution of the illness. The amplitude and latency values of visual evoked potentials were affected in the first and second group of patients. CONCLUSIONS: This neuropathy is of a toxic nutritional-metabolic type and the severity of the clinical-neurophysiological picture is determined by the time of evolution of the symptoms, before the diagnosis of the illness. PMID- 11484144 TI - [Flavonoid drugs as antioxidant in intraocular irrigating solution]. AB - PURPOSE: To study the protective effect for vitrectomy of intraocular irrigating solutions containing different concentrations of two antioxidants: Meralop and Activadone and compare them with saline solutions and BSS plus. MATERIAL AND METHODS: Seventy-five pigmented rabbits were divided into 8 groups: I. Control (n=8); II. Gas-vitrectomy (n=15); III. Vitrectomy + saline solution (n=6); IV. Vitrectomy + BSS plus (n=9); V. Vitrectomy + Activadone 90 mg/l (n=8); VI. Vitrectomy + Activadone 180 mg/l (n=9); VII. Vitrectomy + Meralop 16 mg/l (n=10); VIII. Vitrectomy + Meralop 64 mg/l (n=10). Antioxidant activity in lens and retina was measured evaluating the levels of lipoperoxides malondyaldehide (MDA) and 4-hydroxynonenal (4-HNE) by spectrophotometry. RESULTS: MDA + 4-HNE concentrations in lens and retina of each group (mean and standard deviation in nmol/g) were: Group I= 3.806DE1.043, 38.787DE10.404. Group II= 4.353DE3.626, 41.554DE18.520. Group III= 297.306DE103.73, 156.69DE6.82. Group IV= 29.39DE15.38, 112.02DE50.12. Group V= 42.31DE20.82, 55.44DE42.21. Group VI= 27.66DE9.29, 181.061DE79.39. Group VII= 134.93DE133.52, 76.89DE36.02. Group VIII= 112.91DE60.30, 79.97DE38.005. CONCLUSIONS: All intraocular irrigation solutions containing antioxidant substances produced lower MDA + 4-HNE levels than saline solution. In the retina, Activadone 90 mg/l produces less MDA concentration than BSS plus. PMID- 11484145 TI - [Suture of skin lacerations using LAT gel (lidocaine, adrenaline, tetracaine)]. AB - PURPOSE: Our aim is to assess the effectiveness and possible use od a new gel formulation for a LAT solution in minor lacerations repair and to propose its use for daily practice in surgical repair of skin lacerations near ocular structures like eyelids, eyebrows or forhead. METHOD: Fifty patients presenting skin lacerations of various origins and locations complying with inclusion criteria were topically anesthesized using LAT gel previous to suturing. Concentrations in final gel solution were as follows: Tetracaine 1.0%. Lidocaine 1.5% and Adrenaline 0.1%. Effectiveness of anaesthesia was assessed by means of various pain evaluation scales filled by patients and physician. RESULTS: Lidocaine infiltration was performed if any sign of pain was observed. 277 sutures were performed. Lidocaine infiltration was needed in five patients (10%) all of those evidencing lacerations in hands or fingers; no laceration in any facial area nor scalp needed any supplementary lidocaine infiltration. Of the total 254 sutures not needing infiltration, 19 (7.49%) elicited a painful response. 97% of all sutures placed in face and scalp elicited a less painful response and this difference was statistically significant against locations other than head. An average dose of 0.82 ml. of LAT was used in our study (consisting in 12.3 mg of Lidocaine, 8.2 mg of Tetracaine an 0.82 mg of Adrenalin). CONCLUSION: LAT gel is an effective alternative for the repair of wounds needing minor surgical procedures, it provides anaesthesia for lacerations in face and scalp of children and adults. PMID- 11484146 TI - [Retinal hemorrhages following phacotrabeculectomy. Differential diagnosis]. AB - PURPOSE: We report a case of a patient who underwent uneventful facotrabeculectomy. Right after surgery patient she complained of very poor vision, showing hemorrhage spots scattered throughout the retina. Differential diagnosis is studied. CONCLUSION: An ocular decompression retinopathy should be the first diagnosis; however clinical details and diagnosis tests suggest the ocular ischemic syndrome as the reason of the problem. PMID- 11484147 TI - [Neuroretinitis in an HIV-infected patient]. AB - CASE REPORT: We present a 35 year-old man, suffering from HIV infection with a left neuroretinitis and bilateral multifocal inner retinitis. A systematic approach supported the presumed diagnosis of cat-scratch disease. The patient was treated with oral azytromicine following a full-clinical recovery. DISCUSSION: We discuss the differential diagnosis of this clinical entity, treatment and clinical evolution. PMID- 11484148 TI - [Vision and sex]. PMID- 11484150 TI - The Aberdeen Indian Health Service infant mortality study: design, methodology, and implementation. AB - Of all Indian Health Service areas, the Aberdeen Area has consistently had the highest infant mortality rate. Among some tribes in this area the rate has exceeded 30/ 1000 live birth and half the infant deaths have been attributed to Sudden Infant Death Syndrome,a rate four to five times higher than the national average. The Indian Health Service, Centers for Disease Control and Prevention, National Institute of Child Health and Human Development, and the Aberdeen Area Tribal Chairmen's Health Board collaborated to investigate these high rates with the goals of refining the ascertainment of the causes of death, improving cause specific infant mortality rates and identifying factors contributing to the high rates. Ten of the 19 tribes or tribal communities, representing 66%of the area population, participated in a 4-year prospective case-control study of infants who died after discharge from the hospital. Infant care practices and socio demographic, economic, medical, health care, and environmental factors were examined. The study included parental interviews, death scene investigations, autopsies, neuropathology studies, medical chart abstractions, blood cotinine assays, and a surveillance system for infant deaths. Controls were the previous and subsequent infants born on the case mother's reservation. From December 1,1992 until November 30,1996,72 infant deaths were investigated. This report describes the study methods and the model employed for involving the community and multiple agencies to study the problem of infant mortality among Northern Plains Indians. Data gathered during the investigations are being analyzed and will be published at a later date. PMID- 11484151 TI - Application of the Cross Battery Approach in the assessment of American Indian children: a viable alternative. AB - This article examines current psychometric and testing practices that appear to do a limited job of assessing the intelligence of American Indian individuals. For several reasons, contemporary approaches are found to be inadequate. Unfortunately, these practices are then employed in making educational decisions and placing these same children into Special Education programs. Alternative methods of testing, including the Gf-Gc Cross-Battery Approach, are discussed and evaluated in terms of usefulness in the evaluation of American Indian children and adolescents. The cross-battery method of testing was found to provide more in depth procedures for bypassing both language and cultural differences among American Indian individuals. PMID- 11484152 TI - Applying a cultural models approach to American Indian substance dependency research. AB - The cognitive anthropological method of deriving cultural models from ethnographic discourse analysis is illustrated in relation to case studies yielding nativistic insights regarding American Indian substance dependency and recovery. Discussion focuses on the broader applicability and local community relevance of incorporating cultural models directly into the design and implementation of prevention and intervention programs. Such an approach may benefit local community cultural revitalization efforts while enhancing the cultural relevance and effectiveness of substance abuse programs. PMID- 11484153 TI - The Dream Catcher Meditation: a therapeutic technique used with American Indian adolescents. AB - This article describes a short-term treatment insight-oriented model for American Indian adolescents, called Dream Catcher Meditation. It is aimed at helping clients' express unconscious conflicts and to facilitate differentiation and healthy mutuality. Though its duration can vary, twelve sessions are outlined here. Session descriptions include goals and sample questions. Also included are anecdotal material and reflections about cultural relevancy. PMID- 11484154 TI - Identification and functional consequences of a new mutation (E155G) in the gene for GCAP1 that causes autosomal dominant cone dystrophy. AB - Mutations in the gene for guanylate cyclase-activating protein-1 (GCAP1) (GUCA1A) have been associated with autosomal dominant cone dystrophy (COD3). In the present study, a severe disease phenotype in a large white family was initially shown to map to chromosome 6p21.1, the location of GUCA1A. Subsequent single stranded conformation polymorphism analysis and direct sequencing revealed an A464G transition, causing an E155G substitution within the EF4 domain of GCAP1. Modeling of the protein structure shows that the mutation eliminates a bidentate amino acid side chain essential for Ca2+ binding. This represents the first disease-associated mutation in GCAP1, or any neuron-specific calcium-binding protein within an EF-hand domain, that directly coordinates Ca2+. The functional consequences of this substitution were investigated in an in vitro assay of retinal guanylate cyclase activation. The mutant protein activates the cyclase at low Ca2+ concentrations but fails to inactivate at high Ca2+ concentrations. The overall effect of this would be the constitutive activation of guanylate cyclase in photoreceptors, even at the high Ca2+ concentrations of the dark-adapted state, which may explain the dominant disease phenotype. PMID- 11484156 TI - Female patient showing hypohidrotic ectodermal dysplasia and immunodeficiency (HED-ID). PMID- 11484158 TI - [Cytogenetic analysis on 1058 cases of acute nonlymphocytic leukemia]. AB - OBJECTIVE: To evaluate the karyotypic status in a large series of acute nonlymphocytic leukemia(ANLL) cases. METHODS: A total of 1058 cases of de novo ANLL were studied. Chromosome preparations were made on bone marrow cells using direct method and short-term culture. Karyotypes were analyzed by R-banding in all cases and G-banding in some cases in addition. RESULTS: Six hundred and thirty cases (60%) had clonal chromosomal abnormalities. 25 categories of major karyotypic abnormalities were found. Among them, 11 were the specific chromosomal rearrangements seen in 481 cases, accounting for 76% of the total number of patients with karyotypic abnormalities. Isolate trisomy 8 (21 cases) was the most common numerical abnormality. t(15;17)(211 cases) and t(8;21)(200 cases) were the most frequent structural abnormalities. The 1.1% of M(2), 72% of M(3), 71% of M(4E0), 50% of M(2), 6 % of M(5) and 1.4% of M(2) had t(7;11), t(15;17), inv(16), t(8;21), t/del(11q 23) and t/del(12p) abnormalities, however, the 100% of t(7;11), 100% of t(15;17 ), 100% of inv(16), 88.5% of t(8;21), 83% of t/del(11q23) and 62% of t/del(12p) were detected in patients with M(2), M(3), M(4E0), M(2), M(5) and M(2) subtypes, respectively. CONCLUSION: By conventional banding technique, including R- and G-bandings, 60% of patients with ANLL may be found to have clonal chromosomal abnormalities which are predominantly specific chromosomal rearrangements correlated with specific FAB subtypes. Thus, karyotype is an important indicator for diagnosis and classification of ANLL. PMID- 11484155 TI - A genomewide scan for loci predisposing to type 2 diabetes in a U.K. population (the Diabetes UK Warren 2 Repository): analysis of 573 pedigrees provides independent replication of a susceptibility locus on chromosome 1q. AB - Improved molecular understanding of the pathogenesis of type 2 diabetes is essential if current therapeutic and preventative options are to be extended. To identify diabetes-susceptibility genes, we have completed a primary (418-marker, 9-cM) autosomal-genome scan of 743 sib pairs (573 pedigrees) with type 2 diabetes who are from the Diabetes UK Warren 2 repository. Nonparametric linkage analysis of the entire data set identified seven regions showing evidence for linkage, with allele-sharing LOD scores > or =1.18 (P< or =.01). The strongest evidence was seen on chromosomes 8p21-22 (near D8S258 [LOD score 2.55]) and 10q23.3 (near D10S1765 [LOD score 1.99]), both coinciding with regions identified in previous scans in European subjects. This was also true of two lesser regions identified, on chromosomes 5q13 (D5S647 [LOD score 1.22] and 5q32 (D5S436 [LOD score 1.22]). Loci on 7p15.3 (LOD score 1.31) and 8q24.2 (LOD score 1.41) are novel. The final region showing evidence for linkage, on chromosome 1q24-25 (near D1S218 [LOD score 1.50]), colocalizes with evidence for linkage to diabetes found in Utah, French, and Pima families and in the GK rat. After dense-map genotyping (mean marker spacing 4.4 cM), evidence for linkage to this region increased to a LOD score of 1.98. Conditional analyses revealed nominally significant interactions between this locus and the regions on chromosomes 10q23.3 (P=.01) and 5q32 (P=.02). These data, derived from one of the largest genome scans undertaken in this condition, confirm that individual susceptibility-gene effects for type 2 diabetes are likely to be modest in size. Taken with genome scans in other populations, they provide both replication of previous evidence indicating the presence of a diabetes-susceptibility locus on chromosome 1q24-25 and support for the existence of additional loci on chromosomes 5, 8, and 10. These data should accelerate positional cloning efforts in these regions of interest. PMID- 11484160 TI - [Detection of monosomy 7 or 7q- in cases of myelodysplastic syndrome]. AB - OBJECTIVE: To investigate the value of interphase fluorescence in situ hybridization(FISH) in the detection of monosomy 7 (-7)or deletion of long arm of chromosome 7(7q-) in the cases of myelodysplastic syndrome (MDS). METHODS: Forty six cases of MDS and 10 normal controls were studied simultaneously by conventional karyotype analysis and interphase FISH technique using SpectrumRed directly labeled DNA specific probe for 7q32. Two hundred interphase cells were analyzed for each case and the cells with one red hybridization spot<7% were regarded as positive. RESULTS: Three cases displayed -7/7q- by conventional cytogenetics(CC) and were confirmed by interphase FISH. Six cases in 43 cases who did not show -7/7q- by CC displayed -7/7q- by interphase FISH. CONCLUSION: Interphase FISH is very useful for the detection of -7 or 7q- in MDS and it is more sensitive than CC. PMID- 11484159 TI - [The function of 5'flanking region of mouse neuronal CDC2-like kinase regulatory subunit p35]. AB - OBJECTIVE: To clarify the possible mechanisms which control p35(nck5a) neural specific expression. METHODS: Thirteen luciferase expression vectors which contain different parts of 5'flanking sequence were constructed. Transient expression assay, DNase 1 hypersensitive site(DHSS) assay and primer extension assay were made to reveal the possible mechanisms of p35(nck5a) expression control. RESULTS: It was found that 5'flanking sequence of p35(nck5a) contained 5455bp and there were a few possible promoters in the fragment. The result of transient expression assay in primary cerebrum cortex cell of rat (Neuron) and established non-neuron cell such as HeLa showed the loss of neural-specific expression of all the luciferase expression vectors. The result of DNase 1 hypersensitive site assay revealed a DHSS located at the -400bp upstream of p35(nck5a) in brain but not in liver. The primer extension assay found the putative transcription initiation site near the position of DHSS. CONCLUSION: The results above suggest that there is no cis-element to control the neural-specific expression in the 5'flanking sequence, and the control on the chromosome level may be critical to the neural-specific expression of p35(nck5a). PMID- 11484161 TI - [The genotype analysis of glucose-6-phosphate dehydrogenase deficiency in Yunnan province]. AB - OBJECTIVE: To identify glucose-6-phosphate dehydrogenase (G6PD) gene mutations in 23 patients with G6PD deficiency and to gain further understanding of the molecular and genetic background of G6PD gene in Yunnan province, China. METHODS: The mutations located in exons 2-12 and in parts of introns of G6PD gene were analyzed by amplification refractory mutation system(ARMS), natural and mis-match primer PCR/restrict enzyme, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP ) analysis and automatic DNA sequencing. RESULTS: Among these 23 samples, 5 different point mutations in G6PD gene were identified, and they constituted 5 genotypes. There were 7 Han and 3 Dai patients with G487A mutation, 7 cases with both intron 11 T93C and C1311T mutations, 4 cases with intron 5 636 or 637 T-->del mutation, 1 case with G871A mutation, and 1 case with G487A/T93C/C1311T mutation. Two haplotypes, 93C/1311T and 93C/1311T/487A were identified in Yunnan. A strong association was observed between C1311T and the Nla III restriction site produced by intron 11 T93C. The findings of the investigators on IVS-5 636 or 637T-->del in Chinese, on G871A in mainland of China, and on G487A in the Han people of Yunnan have not been reported previously. CONCLUSION: G6PD deficiency is very heterogenous in Yunnan; G487A is one of the common mutations in that province and may be of different origins. Possibly IVS-11 T93C mutation is of non-African origin. IVS-11 T93C and C1311T might jointly result in G6PD deficiency. The above data on G6PD gene mutation types could be useful for clinical diagnosis, prevention of G6PD deficiency, and researches in the origin and migration of minorities in Yunnan or other regions. PMID- 11484162 TI - [High polymorphism at the human P gene locus in Chinese Han and Tibetan populations]. AB - OBJECTIVE: The authors studied two synonymous sites polymorphism (A355A, G780G) and one intervening sequence polymorphism(IVS13-15) of human P gene in 181 and 171 individuals sampled from Chinese Han and Tibetan populations respectively. METHODS: Genomic DNA was extracted from the ACD-blood specimens collected from 352 healthy unrelated individuals by phenol-chloro methods. The amplification of three fragments was performed using PCR technique; the product was digested with restricted enzyme. Agarose electrophoresis was used for typing the PCR product. RESULTS: There were two alleles at each exon variety locus; the allele frequencies in both A355A and G780G were of significant differences between the two populations ( chi(2)=24.54,chi(2)=37.05,P>0.001). No significant difference was found in allele frequency in IVS13-15 locus between the two populations (chi(2)=2.06,P>0.05). And in each of the populations, no significant difference was noted in the genotype frequency between the male and the female. The test for Hardy-Weinberg equilibrium showed that the genotype distributions observed in the two populations were correspondent with the expected. CONCLUSION: There is a remarkable difference in the distribution of allele frequencies at P gene locus between Chinese Han and Tibetan populations, which is related to the variation in skin pigmentation of these two Chinese population. PMID- 11484163 TI - [Genotyping of HIV resistant alleles in indigenous Tibetan ethnic group of China: low frequency of CCR5triangle32 and high frequency of CCR2b-64I alleles]. AB - OBJECTIVE: To investigate the allelic polymorphism of CCR5triangle32, CCR5m303, CCR2b-64I and SDF1-3'A in Tibetan population in Lasa area of China. METHODS: The genomic DNA samples from 330 Tibetan subjects' whole blood samples were purified by use of QIAgen Blood Kit and identified by PCR or PCR-RFLP analyses. RESULTS: The mutation frequencies of CCR5triangle32 and CCR5m303 alleles were lower than 0.15%, and those of CCR2 b-64I and SDF1-3'A alleles were 29.42% and 19.24% respectively in the study samples. The allelic polymorphisms of the four alleles of Tibetan population were similar to those of Chinese Han population. Genotype distribution of the four alleles was in accordance with Hardy-Weinberg equilibrium. The above results suggest that Tibetan population may be relatively susceptive to HIV-1. CONCLUSION: The Chinese Tibetan may have a lower frequency of CCR5triangle32 and a higher frequency of CCR2b-64I allele, compared with Caucasian. PMID- 11484164 TI - [A study of association between atherothrombotic brain infarction and HLA-DQA1 peptide binding motifs]. AB - OBJECTIVE: To analyze the hereditary susceptibility for HLA-DQA1 alleles according to trait of multiple genes leading to disease in atherothrombotic brain infarction (ABI). METHODS: HLA-DQA1 alleles in ABI patients (n=81) and healthy controls (n=99) were detected by PCR-SSP techniques. RESULTS: (1)The frequency of HLA-DQA1*0301 was significantly higher in ABI patients. (2)The frequency of HLA DQA1*0301 was significantly higher in patients with family history of essential hypertension (EH). (3)The frequency of HLA-DQA1*0103 was significantly lower in ABI patients. CONCLUSION: HLA-DQA1*0301 allele might be a correlative gene with hereditary susceptibility of ABI and HLA-DQA1*0301 allele might be a correlative gene in ABI patients with the family history of essential hypertension, whereas HLA-DQA1*0103 allele might be a protective gene in the patients with ABI. PMID- 11484165 TI - [Relationship between methylenetetrahydrofolate reductase gene polymorphism and diabetic nephropathy]. AB - OBJECTIVE: To investigate the relationship between methylenetetrahydrofolate reductase(MTHFR) gene polymorphism and diabetic nephropathy(DN). METHODS: The technology of polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) was used to detect MTHFR gene C677T mutation in 85 healthy controls, 79 type-2 diabetics without DN, and 82 type-2 diabetics with DN confirmed by urine microalbuminous detection. RESULTS: The frequencies of MTHFR gene variant genotypes and alleles in the patients with DN patients were significantly higher than those in the patients without DN and those in healthy controls (P<0.01). CONCLUSION: MTHFR gene C677T mutation is probably one of the genetic risk factors for diabetic nephropathy in Chinese Hans. PMID- 11484166 TI - [A gene analysis of the low density lipoprotein receptor in Chinese with homozygous familial hyperchol-esterolemia]. AB - OBJECTIVE: To investigate low density lipoprotein receptor(LDLR) gene mutations in Chinese with familial hyperchol-esterolemia(FH). METHODS: Genomic DNA was extracted from five unrelated children with clinical diagnosis of homozygous FH, together with their family members. Promoter and all of the 18 exons of LDLR gene were amplified by polymerase chain reaction (PCR) and were analyzed by single strand conformation polymorphism (SSCP). The PCR products of abnormal patterns shown by SSCP were sequenced directly. RESULTS: Two point mutations (A606T, C263R) were found in two families respectively. The two mutations were only found in China, and C263R is a novel mutation. CONCLUSION: The two mutations of LDLR gene are the cause of FH and there may be LDLR gene mutations only carried by FH patients in China. PMID- 11484167 TI - [Genetic polymorphisms of cytochrome P450 1A1 and susceptibility of early-onset Parkinson's disease]. AB - OBJECTIVE: To assess the possible association between the Msp I polymorphisms of cytochrome P450 1A1(CYP1A1) and the susceptibility of early-onset Parkinson's disease among Hans in the northern part of China. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to analyze three genotypes A, B and C in 3'flanking region of CYP1A1 in 126 patients with idiopathic early-onset Parkinson's disease and 172 healthy controls. RESULTS: The frequencies of genotypes A, B and C in patients were 41.3%, 43.6% and 15.1% while those in the controls were 34.9%, 51.7% and 13.4%, respectively. No statistically significant difference in the frequencies of the three genotypes was observed between the two groups. The frequencies of two alleles were of no significant difference between the patients and controls. CONCLUSION: The above results suggest that the Msp I polymorphisms of cytochrome P450 1A1 itself might not be associated with idiopathic early-onset Parkinson's disease. PMID- 11484168 TI - [Relationship of vitamin D receptor polymorphism with breast cancer]. AB - OBJECTIVE: To detect the association between vitamin D receptor polymorphism and breast cancer. METHODS: Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method was used. Two restriction fragment length polymorphisms in the 3'region of vitamin D receptor (VDR) gene, Taq I and Apa I, were tested for association with breast cancer risk in 86 females with breast cancer and 134 healthy female controls. RESULTS: Allele frequencies of the 3'Taq I polymorphism showed a significant association (P=0.0004, OR:5.39,CI:1.81 17.20). The further study of genotype found the association of Tt, tt with breast cancer. The haplotype analysis of Apa I and Taq I showed a linkage disequilibrium between t-allele and A-allele. The frequency of tA haplotype was higher in breast cancer patients than in controls (P=0.001), indicating that tA haplotype is associated with breast cancer. The alleles and haplotype of the two loci had not any difference among the clinical subgroups. CONCLUSION: There is association between vitamin D receptor polymorphism and breast cancer. PMID- 11484169 TI - [The apolipoprotein E gene polymorphism in children with steroid-resistant idiopathic nephrotic syndrome]. AB - OBJECTIVE: To study apolipoprotein E gene polymorphism in children with steroid- resistant idiopathic nephrotic syndrome (SRINS). METHODS: Sixty children with SRINS (including 9 children with focalsclerosing glomerulonephritis) were observed in comparison with 80 age-sex-matched healthy children. Seven lipoprote in metabolism parameters including serum total-cholesterol(TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein AI(apoAI), apolipoprotein B (apoB), lipoprotein (a)[Lp (a)] were detected by enzyme method, and single-strand conformation polymorphism(PCR-SSCP) was used to determine the apoE genotypes in the two groups. RESULTS: Serum TC, TG, HDL-C, LDL-C, apoAI, apoB, Lp(a) in the observation group were significantly higher than those in the healthy controls (P<0.01). After 6 months, the levels of serum TC, TG, LDL-C,apoB,Lp(a) were 81.25%, 78.13%, 84.38%, 87.50% and 78.13% respectively in 32 SRINS cases of the observation group. The apo epsilon 2 allele gene frequence was higher in the observation group than in the controls (11.66% vs 5.00%, P<0.05). The apo apsilon 2 allele gene frequence in the cases of focalsclerosing glomerulonephritis was significantly higher than that in the controls (22.22% vs 5.00%, P<0.01). CONCLUSION: The above results indicate the obvious and long term abnormalities of serum lipoprotein metabolism in the children with SRINS. The apo apsilon 2 allele gene frequence is higher in the children with SRINS. There are risk factors for atherosclerosis and cardiovascular diseases in the children with SRINS. The antilipemic drugs should be considered in use for the children with SRINS. PMID- 11484170 TI - [Association between angiotensin system gene polymorphism and essential hypertension]. AB - OBJECTIVE: To investigate whether angiotensinogen(AGT) M235T, angiotensin II type I receptor(AT(1)R) gene A1166C and angiotensin converting enzyme(ACE) gene I/D polymorphism are implicated in human essential hypertension(HT) in Chinese. METHODS: Polymerase chain reaction(PCR) and PCR combined with restriction enzyme digestion were used to detect AGT gene M235T, AT(1)R gene A1166C variations and ACE gene I/D polymorphism in 161 hypertensive patients and 134 normotensive controls. RESULTS: No statistically significant differences were found in frequencies of the ACE D allele, AGT gene 235T and AT(1)R A1166C between hypertensive patients and normotensive controls, but in hypertensive patients aged <60 years the frequencies of the ACE D allele and AGT gene 235T were significantly higher than those of the normotensive controls (P<0.05). The analysis of combined genotypes of AGT gene and ACE gene showed that the combined genotypes of DD-TT and ID-TT were significantly higher in hypertensive patients than in normotensive controls. Significant relationships between the ACE genotype and serum ACE activity were found in both groups(P<0.05). CONCLUSION: The ACE D allele and AGT 235T polymorphism may be involved in the early occurrence of HT. The combined genotypes of DD-TT and ID-TT may be a dangerous genetic factor for HT in Chinese. PMID- 11484171 TI - [Studies on association between lipoprotein lipase gene polymorphisms of Pvu II site and hypertriglyceridemics in Chinese]. AB - OBJECTIVE: To explore whether Pvu II restriction fragment length polymorphisms (RFLP) in the lipoprotein lipase(LPL) gene are associated with hypertriglyceridemics (HTG). METHODS: Pvu II restriction fragment length polymorphisms in the lipoprotein lipase gene on a sample of 135 HTG patients and 193 age-matched healthy individuals in Chengdu area were detected with the method of PCR-RFLP. RESULTS: The P+ P+ genotype frequency and P+ allelic frequency of LPL gene for HTG cases are higher than those for control groups (0.460 vs 0.337,P<0.05; 0.689 vs 0.565,P<0.01). The serum levels of TGapoCII, apoCIII, apoE and TG/HDL-C in P+P+ genotype are higher than those in P-P- genotype(P<0.05). CONCLUSION: The results suggest that P+P+ genotype in the lipoprotein lipase gene is associated with susceptibility to hypertriglyceridemics in Chinese population. PMID- 11484172 TI - [No evidence for association between the alpha 2-macroglobulin polymorphism and Alzheimer's disease in the Han Chinese]. AB - OBJECTIVE: To determine the effect of the pentanucleotide deletion polymorphism adjacent to a consensus splice site in intron 17 of alpha 2-macroglobulin(A2M) gene on the development of late-onset Alzheimer's disease (LOAD) in the Han Chinese in Guangzhou area. METHODS: A2M and apolipoprotein E genotypes were examined by polymerase chain reaction(PCR) and restriction fragment length polymorphism (RFLP) techniques in the patients(32 men and 65 women) with LOAD and age-matched healthy elderly Chinese subjects (57 men and 54 women). RESULTS: The A2M gene deletion allele frequency was 2.6% in the cases with LOAD and 2.7% in the controls. In the LOAD cases, the frequency of apoE-epsilon4 increased significantly (Z=3.32, P<0.01). ApoE-epsilon4 was associated with LOAD (RR=2.67, chi(2)=10.71, P<0.01) while the A2M deletion polymorphism was not associated with LOAD even the samples were stratified according to the presence of the apoE epsilon4 allele. CONCLUSION: The above data demonstrate that there is no evidence for the association of A2M polymorphism with the development of LOAD in the Han Chinese population in Gaungzhou. PMID- 11484173 TI - [Effects of phenytoin on structural aberration of human sperm chromosomes in vitro]. AB - OBJECTIVE: To detect the mutagenic effects of phenytoin on human sperm chromosomes. METHODS: The mutagenic effects of phenytoin at 10, 20 and 40 microgram/ml were tested by an in vitro testing system of human sperm chromosomes, with leomycin A5 as a positive control and mutagen-free solution as the blank control. RESULTS: The frequencies of sperm with structural chromosomal aberrations and breakages of sperm chromosomes in the groups of 10 microgram/ml, 20 microgram/ml and 40 ug/ml of phenytoin were all higher than those in the blank control, but only in the group of 40 ug/ml, the elevated frequencies were statistically significant (P<0.005, P<0.05). CONCLUSION: Phenytoin has probably mutagenic potential effect on human sperm cells. PMID- 11484174 TI - [A simple and rapid new method for SNP typing by single-tube bi-directional allele specific amplification]. AB - OBJECTIVE: To establish a new method for single nucleotide polymorphism(SNP) typing based on allele specific PCR: single-tube bi-directional amplification (SB ASA), and study the influence on specific extension by introducing a mismatch at the third 3'terminal base of allele specific primers. METHODS: Two allele specific primers, with a mismatch introduced at the third 3'terminal base, were both included in PCR system; they extended in opposite directions and amplified two allele specific fragments different in size. The genotype was determined by observing the length of amplified fragments after agarose electrophoresis. The proper ranges of annealing temperature (Ta) under which primers can specifically extend were achieved by observing the amplification status at different temperatures. RESULTS: SB-ASA was successfully used to type 36 samples for four different kinds of SNPs. Typing results were completely consistent with those by directional sequencing. Proper Ta ranges of two primers were expanded respectively from 64-69 degrees centigrade to 46-66 degrees centigrade and from 60-62 degrees centigrade to 56-61 degrees centigrade by introducing a mismatch at the third 3'terminal base. CONCLUSION: SB-ASA is a simple, rapid and efficient new method for SNP typing. During allele specific PCR reaction, specific primers with a mismatch at the third 3'terminal base have more power to identify two alleles. PMID- 11484175 TI - [Phenotyping and genotyping studies in a family with the compound heterozygosity for a deletional delta beta-thalassemia and a beta-thalassemia]. AB - OBJECTIVE: To investigate the relationship between genotype and phenotype of a deletional delta beta-thalassemia and explore an approach to rapid prenatal diagnosis for compound heterozygotes of this defect and a beta-thalassemia. METHODS: A total of ten members in a Chinese family who had a 5-year-old propositus with thalassemia major and requested prenatal diagnosis for the second pregnancy were investigated. For genotyping analysis, the deletional delta beta thalassemia was characterized by gap-PCR method and beta-thalassemia mutations were defined by reverse dot blot(RDB). For phenotyping analysis, hematological data including the RBC indices, quantification of HbF and HbA2 by Hb electrophoresis and the ratio of globin chain Ggamma/(Ggamma+Agamma) analyzed by capillary electrophoresis (CE) were obtained. RESULTS: The propositus inherited her mother's delta beta-thalassemia gene mutation and her father's CD41-42(-CTTT) frameshift mutation. Of nine members in this family screened for this type of deletion, four were positive and the phenotype could be explained satisfactorily by genotype. The results of prenatal diagnosis showed that the fetus was normal and had no beta-globin gene defects in both chromosomes. CONCLUSION: This is the first time to have performed prenatal diagnosis in Chinese family at-risk of compound heterozygotes for beta-thalassemia and delta beta-thalassemia in mailand China. The strategy to analyze the disease presented here may be a valuable reference to the similar problem. PMID- 11484176 TI - [Cloning of the mouse Doc-1R gene by genomic walking]. AB - OBJECTIVE: To obtain the genomic sequences of the mouse Doc-1R gene. METHODS: Gene-specific primers were designed and synthesized based on the cDNA sequences of the mouse Doc-1R gene. With the use of genomic walking strategy, the mouse genomic walking library was amplified by the polymerase chain reaction(PCR). Mouse genomic library constructed with a special adaptor was utilized as a template to amplify the desired fragment by nested PCR. RESULTS: A desired fragment of 1.5 kb was obtained. Sequence analysis of the desired fragment confirmed that the genomic cloning of the Doc-1R gene was successful. This gene contains four exons and three introns. All of the splice donor/acceptor site sequences are in accordance with the consensus 'GT-AG' rule. CONCLUSION: The genomic walking strategy is simple, efficient and reliable; it is an ideal method of cloning genomic fragments. PMID- 11484177 TI - [A rapid and simple detection of DNA fragment with point mutation by capillary electrophoresis]. AB - OBJECTIVE: To establish a rapid and simple method with high efficiency in detecting point mutation of genomic DNA. METHODS: Four DNA fragments that were different from each other in only one based were amplified by using primers with artificial point mutation based on the sequence of exon 7 of p53 gene, and then were separated by capillary electrophoresis(CE). The neutral coated capillary and 4% linear polyacrylamide gel buffer were used, and the wave length of ultraviolet detector was 254 nm. RESULTS: A homozygous 196 bp DNA fragment can be separated into one dsDNA peak and two ssDNA peaks within 25 minutes, and the heterozygous 196 bp DNA fragments that were made with mixed wild type and mutated DNA can be separated into one dsDNA peak and three ssDNA peaks. The three ssDNA fragments that differ in one nucleotide can be easily separated with good resolution. CONCLUSION: CE technique is rapid, sensitive, accurate and well reproducible. It is an efficient and reliable method for rapidly screening point mutation. PMID- 11484178 TI - Whisker-reinforced bioactive composites containing calcium phosphate cement fillers: effects of filler ratio and surface treatments on mechanical properties. AB - Calcium phosphate cement (CPC) sets to form microporous solid hydroxyapatite with excellent osteoconductivity, but its brittleness and low strength prohibit use in stress-bearing locations. The aim of this study was to incorporate prehardened CPC particles and ceramic whiskers in a resin matrix to improve the strength and fracture resistance, and to investigate the effects of key microstructural variables on composite mechanical properties. Two types of whiskers were used: silicon nitride, and silicon carbide. The whiskers were surface-treated by fusing with silica and by silanization. The CPC particle fillers were either silanized or not silanized. Seven mass ratios of whisker-silica/CPC were mixed: 0:1 (no whisker-silica), 1:5, 1:2, 1:1, 2:1, 5:1, and 1:0 (no CPC). Each powder was blended with a bisphenol-a-glycidyl methacrylate-based resin to harden in 2 x 2 x 25 mm molds by two-part chemical curing. The specimens were tested in three-point flexure to measure strength, work-of-fracture (toughness), and elastic modulus. Two-way analysis of variance was used to analyze the data, and scanning electron microscopy was used to examine specimen fracture surfaces. The whisker-silica/CPC ratio had significant effects on composite properties (p < 0.001). When this ratio was increased from 0:1 to 1:0, the strength was increased by about three times, work-of-fracture by five times, and modulus by two times. Whisker surface treatments and CPC filler silanization also had significant effects (p < 0.001) on composite properties. Scanning electron microscopy revealed rough fracture surfaces for the whisker composites with steps and whisker pullout. Resin remnants were observed on the surfaces of the pulled-out whiskers, indicating strong whisker-matrix bonding. In conclusion, incorporating highly osteoconductive CPC fillers and ceramic whiskers yielded composites with substantially improved mechanical properties compared with composites filled with CPC particles without whiskers. The composite properties were determined by whisker-to-CPC ratio and filler surface treatments. PMID- 11484179 TI - Implantation of octacalcium phosphate combined with transforming growth factor beta1 enhances bone repair as well as resorption of the implant in rat skull defects. AB - In our previous study, we reported that synthetic octacalcium phosphate (OCP) enhances bone repair if implanted in rat skull defects. We hypothesized that OCP can be used as an effective carrier for transforming growth factor-beta1 (TGF beta1) to promote bone repair. We designed the present study to investigate histomorphometrically whether combination with recombinant human TGF-beta1 could promote bone repair caused by OCP per se (Control/OCP). A full-thickness standardized trephine defect was made in the rat parietal bone and OCP combined with recombinant human TGF-beta1 (TGF-beta1/OCP) or Control/OCP was implanted into the defect. Four rats from each group were fixed at 2, 4, and 8 weeks after implantation. Histomorphometrical analysis of the percentage of newly formed bone (n-Bone %) and remaining implants (r-Imp %) in the defect was performed. The statistical analysis showed the n-Bone % of TGF-beta1/OCP was significantly higher than that of the Control/OCP in week 4, whereas the r-Imp % of TGF beta1/OCP was significantly lower than that of the Control/OCP. The present study demonstrated that OCP can be used as an effective carrier for TGF-beta1 and their combination enhances bone repair as well as resorption of the carrier OCP in the early stage of bone formation. PMID- 11484180 TI - Cell behavior on laser surface-modified polyethylene terephthalate in vitro. AB - This work has been undertaken to study the cell behavior of L929 fibroblasts on the laser irradiated polyethylene terephthalate (PET) surface. To modify the surface properties of the PET, CO2 pulsed laser at the wavelength of 9.25 microm and KrF excimer laser at 248 nm with various number of pulses were used. Laser irradiation caused some changes in the chemical and physical properties of the laser-treated film surfaces, which were evaluated using different techniques. These changes may affect the cell adhesion and growth on the laser-treated PET. Therefore, cell attachment and spreading were investigated on the laser-treated PET in vitro. The data from in vitro assays showed the fibroblast cells were attached and proliferated extensively on the CO2 and KrF laser-treated films in comparison with the unmodified PET. The results obtained from the cell behavior studies revealed that surface morphology and wettability affected cell adhesion and spreading on the laser-treated PET. PMID- 11484181 TI - Effect of material geometry on cartilagenous tissue formation in vitro. AB - The effect of material geometry, as defined by average pore size, on chondrocyte phenotype and cartilagenous tissue formation in vitro was examined. Bovine articular chondrocytes were plated on porous titanium alloy (Ti6Al4V) discs of different average pore sizes (13, 43, and 68 microm) and grown in culture for 4 weeks. Chondrocyte phenotype was maintained as indicated by the synthesis of large proteoglycans (Kav +/- SD: 13 microm = 0.28 +/- 0.01; 43 microm = 0.29 +/- 0.01; 68 microm = 0.27 +/- 0.02) and type II collagen. Light microscopical examination of histological sections of the composites showed that cartilagenous tissue had formed on all discs. The cartilagenous tissue on the discs of the smallest average pore size (13 microm) was significantly thicker than the tissue on the discs of larger average pore sizes and also had greater amounts of proteoglycan [mean glycosaminoglycan content +/- SD microg/disc): 13 microm = 246.9 +/- 7.8; 43 microm = 190.4 +/- 10.2; 68 microm = 156.6 +/- 25.8, p = 0.002] and DNA [mean DNA content +/- SD microg/disc): 13 microm = 12.5 +/- 0.6; 43 microm = 8.3 +/- 0.2; 68 microm = 9.3 +/- 0.9, p = 0.0008]. However, the amount of proteoglycan accumulated per cell was similar in the tissues generated on the discs of different average pore sizes. In contrast, the amount of collagen in the cartilagenous tissues showed no significant differences between the different pore sizes, but the amount of collagen accumulated per cell was less in the tissue formed on the smallest pore size disc (13 microm) as compared with the tissue formed on the discs of the larger pore sizes [mean hydroxyproline content/DNA (microg/microg) +/- SD: 13 microm = 1.56 +/- 0.2; 43 microm = 2.19 +/ 0.2; 68 microm = 2.3 +/- 0.3]. These results suggest that material geometry, as defined by pore size, can affect the amount and composition of the cartilagenous tissue that forms. PMID- 11484182 TI - Growth inhibition of cultured smooth muscle cells by corrosion products of 316 L stainless steel wire. AB - The potential cytotoxicity on vascular smooth muscle cells of corrosion products from 316 L stainless steel, one of most popular biomaterials of intravascular stents, has not been highlighted. In this investigation, 316 L stainless steel wires were corroded in Dulbecco's modified eagle's medium with applied constant electrochemical breakdown voltage, and the supernatant and precipitates of corrosion products were prepared as culture media. The effects of different concentrations of corrosion products on the growth of rat aortic smooth muscle cells were conducted with the [3H]-thymidine uptake test and cell cycle sorter. Both the supernatant and precipitates of corrosion products were toxic to the primary culture of smooth muscle cells. The growth inhibition was correlated well with the increased nickel ions in the corrosion products when nickel concentration was above 11.7 ppm. The corrosion products also changed cell morphology and induced cell necrosis. The cell growth inhibition occurred at the G0/G1 to S transition phase. Similar to our recent study of nitinol stent wire, the present investigation also demonstrated the cytotoxicity of corrosion products of 316 L stainless steel stent wire on smooth muscle cells, which might affect the poststenting vascular response. PMID- 11484183 TI - In vivo evaluation of a novel porous hydroxyapatite to sustain osteogenesis of transplanted bone marrow-derived osteoblastic cells. AB - Biosynthetic bone grafts are considered to contain one or more of three critical components: osteoprogenitor cells, an osteoconductive matrix, and osteoinductive growth factors. The basic requirements of the scaffold material are biocompatibility, mechanical integrity, and osteoconductivity. A major design problem is satisfying these requirements with a single composite. In this study, we hypothesize that one composite that combines bone marrow-derived osteoblasts and a novel mechanical reinforced porous hydroxyapatite with good biocompatibility and osteoconductivity (HA/BMO) can reach these requirements. A novel sintered porous hydroxyapatite (HA) was prepared by the following procedures. The HA slurry was foamed by adding polyoxyethylenelaurylether (PEI) and mixing. The pores were fixed by crosslinking PEI with diepoxy compounds and the HA porous body was sintered at 1200 degrees C for 3 h. The HA sintered porous body had a high porosity (77%), and was completely interconnected. Average pore diameter was 500 microm and the interconnecting path 200 microm in diameter. The compressive (17 MPa) and three-point bending (7 MPa) strengths were high. For in vivo testing, the 2-week subcultured HA/BMO (+) composites were implanted into subcutaneous sites of syngeneic rats until 8 weeks after implantation. These implants were harvested at different time points and prepared for the biochemical analysis of alkaline phosphatase activity (ALP) and bone osteocalcin content (OCN), and histological analysis. ALP and OCN in the HA/BMO group were much higher than those in the HA without BMOs control group 1 week after implantation (p < 0.001). Light microscopy revealed mature bone formation in the HA/BMO composite 4 weeks after implantation. In the SEM study, mineralized collagenous extracellular matrix was noted in HA/BMO composite 2 weeks after implantation with numbers of active osteoblasts. We conclude that the composite of the novel HA and cultured BMOs has osteogenic ability in vivo. These results provide a basis for further studies on the use of this composite as an implant in orthopaedic surgery. PMID- 11484184 TI - Attachment of fibronectin to poly(vinyl alcohol) hydrogels promotes NIH3T3 cell adhesion, proliferation, and migration. AB - Hydrogels have been used in biology and medicine for many years, and they possess many properties that make them advantageous for tissue engineering applications. Their high water content and tissue-like elasticity are similar to the native extracellular matrix of many tissues. In this work, we investigated the potential of a modified poly(vinyl alcohol) (PVA) hydrogel as a biomaterial for tissue engineering applications. First, the ability of NIH3T3 fibroblast cells to attach to PVA hydrogels was evaluated. Because of PVA's extremely hydrophilic nature, important cell adhesion proteins do not adsorb to PVA hydrogels, and consequently, cells are unable to adhere to the hydrogel. By covalently attaching the important cell adhesion protein fibronectin onto the PVA hydrogel surface, the rate of fibroblast attachment and proliferation was dramatically improved, and promoted two-dimensional cell migration. These studies illustrate that a fibronectin-modified PVA hydrogel is a potential biomaterial for tissue engineering applications. PMID- 11484185 TI - Effect of RGD coating on osteocompatibility of PLGA-polymer disks in a rat tibial wound. AB - Osteocompatibility of porous polylactic-glycolic acid (PLGA) disks coated with synthetic peptides was assessed in 5-mm diameter unicortical tibial osseous wounds in rats. The coatings consisted of various ratios of peptides including the tripeptide arginine-glycine-aspartic acid (RGD) and the inactive arginine glycine-glutamic acid (RGE). When left empty, the tibial wounds healed spontaneously with proliferation of intramedullary woven bone within 1 week. The reactive bone was resorbed, and by 3 weeks, the cortical wound was healed with lamellar bone, and the medullary space was repopulated with marrow. When PLGA disks were implanted there was a delay in repair with reduced bone fill and no bone bridging at 3 weeks. When disks were coated with increasing amounts of RGD peptide, there was a biphasic effect on osteocompatibility and on osseous ingrowth. Evaluation at 10 days showed a dose-dependent increase, with 1.5-fold greater osteocompatibility (p < 0.05) and 1.6-fold more osseous ingrowth into the polymer (p < 0.01) than uncoated disks. With more RGD and with undiluted RGE, osteocompatibility and osseous ingrowth were the same as with uncoated disks. At 3 weeks, there were no significant differences among all the groups. These data indicate that RGD coating enhanced early stages of osteocompatibility and ingrowth. PMID- 11484186 TI - Cell response to herpes simplex virus type 1 infection mediated by biphasic calcium-phosphate ceramics: in vitro approach. AB - Based on well-documented data showing that bioactive ions (such as Ca2+, PO4-, etc.) released by BCPC induce various cell responses and on the significance of herpes outbreaks in human pathology, we investigated whether BCPC can modify cell response to HSV-1 infection. The roles of some physical and chemical properties of ceramics were evaluated using three BCPC samples--French commercial macro microporous (FR) and two Bulgarian microporous laboratory samples--one of which was modified with magnesium (BG and BG + Mg). Samples only washed in 0.9% NaCl were designated as nonconditioned while those resuspended in cell growth medium for 10 days after washing were designated as conditioned. Experiments were done on cells from the continuous MDBK line precultured on BCPC surfaces for different time intervals and thereafter HSV-1 infected. The yield of infectious virus progeny, measured as virus titers, was the parameter used to determine the cell response to HSV-1 infection mediated by BCPC as compared to that of the virus control, that is, virus yield in cells cultured without BCPC. The data obtained show that all three nonconditioned BCPC samples were able to modify cells to resist HSV-1 infection. The prolongation of the resistant state depended on the specific physical and chemical properties of the particular BCPC sample: as the data show that the conditioning procedure (1) increased the ability of BG + Mg to promote cell resistance, and (2) reduced the ability of FR samples to modify cells to resist HSV-1 infection. The data obtained show that apart from Ca2+ and PO4-, ions of biometals such as Mg2+ also are responsible for the induction and maintenance of cell resistance to HSV-1 infection. PMID- 11484187 TI - Respiratory burst response of peritoneal leukocytes adhering to titanium and stainless steel. AB - Titanium sheets, made hydrophilic by oxidative cleaning or hydrophobic by treatment with butanol, and stainless steel sheets with different patterns of pores (straight phi = 0.8 mm) were implanted into the peritoneal cavity of mice. The implants were removed after 2 h, and the surface-adhering leukocytes were stained with propidium iodide and fluorescein diacetate to quantitate cell adhesion and to indicate the presence of leaks in the cell membrane. The ability of the surface-adhering leukocytes to mount a respiratory burst response after stimulation with PMA or zymosan was measured by chemiluminiscence. The results show that stainless steel without pores induces membrane leakage in 80% of the surface-adhering leukocytes compared with 65% of cells adhering to porous steel. Hydrophilic titanium induces membrane leakage in 48% of the surface-adhering leukocytes compared with 19% of cells adhering to hydrophobic titanium. The respiratory burst response of the surface-adhering leukocytes stimulated with PMA was attenuated on stainless steel and hydrophilic titanium compared with hydrophobic titanium. Thus, butanol treatment of titanium and pores in stainless steel increase the biocompatibility of the materials. PMID- 11484188 TI - Poly(L-lactide)acid/alginate composite membranes for guided tissue regeneration. AB - The barrier membranes for guided tissue regeneration (GTR) to treat bone defects have to satisfy the criteria of biocompatibility, cell-occlusiveness, space making, tissue integration and clinical manageability. In this study a system constituted of a poly(L-lactide) acid (PLLA) asymmetric membrane combined with an alginate film was prepared. The PLLA membrane functions to both support the alginate film and separate the soft tissue; the alginate film is intended to act as potential vehicle for the growth factors to promote osteogenesis. The structural, morphological, and mechanical properties of the bilamellar membrane and its stability in culture medium were evaluated. Moreover, the feasibility of using the alginate membranes as controlled-release delivery vehicles of TGF-beta was monitored. Finally, the bacterial adhesion and permeability of Streptococcus mutans, selected for the high adhesive affinity, were monitored. The results showed that the surfaces of the alginate side, to be used in contact with the bone defect, were rougher than PLLA ones. When in contact with complete culture medium, the PLLA-alginate membrane retained its mechanical and structural properties for more than 100 days. Then, the degradation processes occurred but the membrane continued to be stable and manageable for 6 months. Growth factors such as TGF-beta can be incorporated into alginate membranes functioning as drug delivery vehicle, and retain the biological activity when tested in an in vitro model system. The obtained membrane acted as a barrier to the passage of S. mutans bacteria and showed to promote a lower bacterial adhesion with respect to commercial GTR membranes. PMID- 11484189 TI - Hydroxylapatite binds more serum proteins, purified integrins, and osteoblast precursor cells than titanium or steel. AB - The implant material hydroxylapatite (HA) has been shown in numerous studies to be highly biocompatible and to osseointegrate well with existing bone; however, the molecular mechanisms at work behind this osseointegration remain largely unexplored. One possibility is that the implant, exposed to the patient's blood during surgery, adsorbs known cell adhesive proteins such as fibronectin and vitronectin from the serum. Osteoblast precursors could then adhere to these proteins through integrin-mediated mechanisms. In the present study, we have used a quantitative ELISA assay to test the hypothesis that hydroxylapatite will adsorb more fibronectin and vitronectin from serum than two commonly used hard tissue materials, commercially pure titanium, and 316L stainless steel. We further used the ELISA, as well as a standard cell adhesion assay, to test the hypothesis that increased protein adsorption will lead to better binding of purified integrins alpha5beta1 and alpha(v)beta3 and osteoblast precursor cells to the HA than to the metals. Our results show that fibronectin, vitronectin, alpha5beta1, alpha(v)beta3, and osteoblast precursor cells do indeed bind better to HA than to the metals, suggesting that improved integrin-mediated cell binding may be one of the mechanisms leading to better clinical bone integration with HA coated implants. PMID- 11484190 TI - Sodium alginate sponges with or without sodium hyaluronate: in vitro engineering of cartilage. AB - Studies are underway to design biosystems containing embedded chondrocytes to fill osteochondral defects and to produce a tissue close to native cartilage. In the present report, a new alginate three-dimensional support for chondrocyte culture is described. A sodium alginate solution, with or without hyaluronic acid (HA), was freeze-dried to obtain large-porosity sponges. This formulation was compared with a hydrogel of the same composition. In the sponge formulation, macroscopic and microscopic studies demonstrated the formation of a macroporous network (average pore size, 174 microm) associated with a microporous one (average pore size, 250 nm). Histological and biochemical studies showed that, when loaded with HA, the sponge provides an adapted environment for proteoglycan and collagen synthesis by chondrocytes. Cytoskeleton organization was studied by three-dimensional fluorescence microscopy (CellScan EPR). Chondrocytes exhibit a marked spherical shape with a nonoriented and sparse actin microfilament network. Type II collagen was detected in both types of sponges (with or without HA) using immunohistochemistry. In conclusion, the sponge formulation affords new perspectives with respect to the in vitro production of "artificial" cartilage. Furthermore, the presence of hyaluronate within the alginate sponge mimics a functional environment, suitable for the production by embedded chondrocytes of an extracellular matrix. PMID- 11484191 TI - Studies on in vitro evaluation for the biocompatibility of various biomaterials: inhibitory activity of various kinds of polymer microspheres on metabolic cooperation. AB - Gap junctional intercellular communication is a function that plays an important role in maintaining cell and tissue homeostasis and in regulating cell growth, development, and differentiation. Change in this function when contacting fibroblasts with various polymer microspheres was estimated using the metabolic cooperation assay system. When the cells were in contact with the microspheres after their adhesion onto a substrate, the function did not alter. However, when they were in contact with precoated microspheres on test dishes, the function was inhibited as the quantity of microspheres increased. Moreover, the inhibition level increased as the diameters of polyethylene and polystyrene microspheres decreased. However, no inhibition was observed if precoated microspheres were composed from poly(L-lactic acid). These findings suggest that the size and the material of microspheres, and how cells recognize the microspheres, are factors affecting cell function of gap junctional intercellular communication. Therefore, estimating this function may provide valuable information about the biocompatibility of many kinds of materials even in the form of particles. PMID- 11484192 TI - Ellipsometric in vitro studies on blood plasma and serum adsorption to zirconium. AB - Ellipsometry/antibody techniques were used to study the adsorption of heparinized human blood plasma and serum onto spontaneously oxidized zirconium, and a colorimetric assay measured the formation of kallikrein by the surface in citrated plasma. After 10 min incubation in the blood plasma the protein film thickness was approximately 4.2 nm, and the film bound polyclonal antibodies mainly against high molecular weight kininogen (HMWK), immunoglobulin G (IgG) and fibrinogen. After 5 or 60 min of incubations in whole normal or EGTA sera, antibodies against complement factor 3 (C3) and complement factor 3d (C3d) deposited to the surface. Factor H and complement factor 1q (C1q) were detected similarly after 1 and 5 min of incubation in 1-10% normal serum in veronal buffer, respectively. The indications are that upon contact with blood plasma, zirconium activates the intrinsic pathway of coagulation and is opsonized with C3. The failure to detect properdin and transient presence of factor H at the surface suggest that complement binds to zirconium although the activation becomes quickly down-regulated. PMID- 11484193 TI - Bone bonding in bioactive glass ceramics combined with a new synthesized agent TAK-778. AB - We studied the stimulatory effects of TAK-778, a new synthetic 3-benzothiepin derivative that promotes osteoblast differentiation, in the bonding of bone to bioactive glass ceramic implants in rabbit tibiae. Smooth-surfaced, rectangular plates (15 x 10 x 2 mm) made of apatite-wollastonite-containing glass ceramic were implanted bilaterally into the proximal metaphyses of rabbit tibiae. Sustained-release microcapsules containing TAK-778 were packed into the medullary cavity in one limb and untreated microcapsules were packed into the contralateral limb to serve as a paired control. At 4, 8, and 16 weeks after implantation, bonding at the bone/implant interfaces was evaluated using a detaching test and histological examination of undecalcified specimens. The tensile failure load increased during weeks 4 to 16 in both groups; the tensile failure load in the TAK-778-treated group was significantly greater than that in the control group at each interval after implantation. Histologically, the TAK-778-treated specimens showed greater active new bone formation mainly in the medullary cavity and more extensive bonding between the implant and bone than the untreated specimens. The results of this study suggest that adding the bone formation-promoting TAK-778 to bioactive glass ceramic implants may significantly accelerate bone apposition to the implants and improve the bonding process at the interface. This would help to establish earlier and stronger bonding of orthopedic ceramic implants to the surrounding bone tissue. PMID- 11484194 TI - Effect of chitosan-polyvinyl pyrrolidone hydrogel on proliferation and cytokine expression of endothelial cells: implications in islet immunoisolation. AB - Earlier we have shown the suitability of chitosan-polyvinyl pyrrolidone (PVP) hydrogel for islet immunoisolation and its inability to activate macrophages. Biomaterials that support vascularization without activating immune competent endothelial cells are desirous in islet immunoisolation. The aim of the present study was to evaluate effect of chitosan-PVP hydrogel on proliferation and activation of endothelial cells. Hydrogel did not allow the majority of cells to adhere well but maintained their viability. Hydrogel leachouts were nontoxic to the cells, as confirmed by tetrazolium reduction (MTT) and Neutral red uptake assays. Exposure to leachouts also did not alter their functionality as seen from normal expression of von Willebrand factor. 3H-thymidine incorporation revealed that hydrogel leachouts did not induce endothelial cell proliferation. Cells cultured on hydrogel and polystyrene control showed comparable expression of interleukin (IL) 6, IL-10, and transforming growth factor beta, with higher expression of tumor necrosis factor alpha as determined by reverse transcription polymerase chain reaction. Taken together these results point out that hydrogel is compatible with endothelial cells and maintains their nonactivated status and hence is suitable as immunoisolation matrix. PMID- 11484195 TI - Comparative effects of nacre water-soluble matrix and dexamethasone on the alkaline phosphatase activity of MRC-5 fibroblasts. AB - In this study we demonstrate, for the first time, that dexamethasone and BMP-2 stimulated alkaline phosphatase (ALP) activity in MRC-5 fibroblasts, a cell line derived from human fetal lung. Previously we reported that the water-soluble matrix (WSM) of nacre obtained from the inner shell layer of the oyster Pinctada maxima, promoted an increase in ALP activity that was dose-dependent. In this work, we show that the effect of WSM is also time-dependent. As a comparison, the effect of WSM was also tested in bone marrow stromal cells because marrow and other bone surface-derived osteoblast stem cells have the inherent direct potential for osteogenesis. WSM promotes cell proliferation and ALP activity when tested with bone marrow cells in concentrations between 135 and 540 microg protein/mL. The effect of WSM on ALP activity of bone marrow stromal cells is similar to that obtained by dexamethasone. These results imply that MRC-5 fibroblasts respond to differentiating factors that promote osteoblastic phenotype in bone-derived cell cultures. PMID- 11484196 TI - Silica sol-gel for the controlled release of antibiotics. I. Synthesis, characterization, and in vitro release. AB - Room temperature processed silica sol-gel (xerogel) was investigated as a novel controlled release carrier of antibiotics (vancomycin). Xerogel characteristics, in vitro release properties, and bactericidal efficacy of the released antibiotic were determined. The xerogel/vancomycin composite showed a long-term sustained release (up to 6 weeks). In addition, bactericidal efficacy of released vancomycin was retained. The kinetics of release and the amount released were dose dependent. The initial, first-order release was followed by a near-zero order release. The time to transition from the first- to zero-order release increased with vancomycin load (from 2 to 3 weeks with load increase from 2.2 to 11.1 mg/g). Regardless of the load, about 70% of the original vancomycin content was released by the transitional point, and the cumulative release after 6 weeks of immersion was about 90%. This study, combined with other reports documenting biocompatibility and controlled resorbability of the xerogel/drug composite in vivo, suggests that silica xerogel is a promising controlled release material for the treatment of bone infections. PMID- 11484197 TI - Pierre Robin sequence and interstitial deletion 2q32.3-q33.2. AB - Pierre Robin sequence (PRS) consists of the nonrandom association of micrognathia, cleft palate (CP), and glossoptosis. It also includes respiratory and feeding difficulties that appear to be neurogenic rather than mechanical in causation. Genetic determinants are thought to underlie this functional and morphological entity, based on the existence of Mendelian syndromes with PRS, and the rare observations of familial nonsyndromic PRS, in which some of the affected individuals have isolated CP. We report the association of PRS with deletion 2q32.3-q33.2 due to an unbalanced reciprocal translocation 46,XX, t(2;21), del 2(q32.3q33.2), and we refine the deletion interval with regard to YAC probes and polymorphic DNA markers. The deletion was shown to be flanked by D2S369 (telomeric) and D2S315 (centromeric), thus it maps to a recently determined chromosomal region known to be nonrandomly associated with CP. This observation supports the hypothesis for the genetic bases of nonsyndromic PRS, strengthens its possible genetic association with isolated CP, and provides a candidate PRS locus, in chromosomal region 2q32.3-q33.2. PMID- 11484198 TI - Distal deletion, del(2)(q33.3q33.3), in a patient with severe growth deficiency and minor anomalies. AB - We report on an 18-month-old boy with a 2q33.3 deletion. The clinical findings observed in the propositus included minor anomalies of face and distal limbs, intrauterine and postnatal growth retardation, microcephaly and, so far, moderate developmental delay. Conventional GTG banded chromosome analysis indicated a small deletion in distal 2q. Subsequent analysis by fluorescent in situ hybridization (FISH) using different probes allowed us to narrow down the deletion to most or all of segment 2q33.3. This case shows the importance of the application of different YAC probes for a precise determination of breakpoints in small interstitial deletions. PMID- 11484199 TI - Hirschsprung disease in an infant with a contiguous gene syndrome of chromosome 13. AB - Hirschsprung disease is a developmental disorder resulting from the arrest of the craniocaudal migration of enteric neurons from the neural crest along gastrointestinal segments of variable length; see Behrman [Nelson textbook of pediatrics, 1992:954-956]. It is a heterogeneous disorder in which familial cases map to at least three loci whose function is necessary for normal neural crest derived cell development. Homozygous mutations in the endothelin-B receptor gene (EDNRB) on 13q22 have been identified in humans and mice with Hirschsprung disease type 2 (HSCR2). The auditory pigmentary disorder, Waardenburg-Shah syndrome, comprises Waardenburg syndrome and Hirschsprung disease and has also been mapped to the EDNRB locus. Hirschsprung disease, malrotation, isochromia, a profound sensorineural hearing loss, and several other anomalies were found in an infant with an interstitial deletion of 13q, suggesting the existence of a contiguous gene syndrome involving developmental genes necessary for the normal growth of the neural crest derivatives of the eye, inner ear, and colon. We report on an additional patient with a deletion in 13q and Hirschsprung disease. Congenital anomalies associated with deletions of the distal long arm of chromosome 13 are sufficiently consistent to suggest a clinical syndrome. PMID- 11484200 TI - Ritscher-Schinzel cranio-cerebello-cardiac (3C) syndrome: report of four new cases and review. AB - Ritscher-Schinzel syndrome, also known as the 3C syndrome, is a rare, autosomal recessive syndrome characterized by craniofacial, cerebellar, and cardiac anomalies. Cardiac manifestations include ventricular septal defect, atrial septal defect, tetralogy of Fallot, double outlet right ventricle, hypoplastic left heart, aortic stenosis, pulmonic stenosis and other valvular anomalies. Central nervous system anomalies include Dandy-Walker malformation, cerebellar vermis hypoplasia and enlargement of the cisterna magna. Craniofacial abnormalities seen are cleft palate, ocular coloboma, prominent occiput, low-set ears, hypertelorism, down-slanting palpebral fissures, depressed nasal bridge and micrognathia. Dandy-Walker malformation, posterior fossa cyst, hydrocephalus and congenital heart defect are common malformations that may occur in isolation or as a part of many syndromes. Accurate genetic diagnosis and counseling require detailed analysis of the external as well as the internal anatomy and knowledge of the relative frequencies of various malformations in syndromes that may have overlapping clinical signs. We have had the opportunity recently to study four cases of the Ritscher-Schinzel syndrome. A review of all reported cases is presented and an attempt made to define the minimum diagnostic criteria for the Ritscher-Schinzel syndrome. Of the nine craniofacial anomalies commonly reported as a part of the Ritscher-Schinzel syndrome, we consider two i.e., cleft palate and ocular coloboma, to be readily and objectively ascertainable. The other seven craniofacial traits, however, are somewhat subjective, require expert interpretation and are sometimes difficult to ascertain in a newborn or stillborn fetus. These are prominent forehead, prominent occiput, hypertelorism, down slanting palpebral fissures, low-set ears, depressed nasal bridge and micrognathia. At least four of these were present in all cases that had a secure diagnosis of the Ritscher-Schinzel syndrome. Thus, the criteria we propose to establish the diagnosis of the Ritscher-Schinzel syndrome in a chromosomally normal sporadic case are the presence of cardiac malformation other than isolated patent ductus arteriosus, cerebellar malformation, and cleft palate or ocular coloboma or four of the following seven findings: prominent forehead, prominent occiput, hypertelorism, down-slanting palpebral fissures, low-set ears, depressed nasal bridge, and micrognathia. PMID- 11484201 TI - Phenotype of five patients with Greig syndrome and microdeletion of 7p13. AB - Here we describe five patients with Greig cephalopolysyndactyly syndrome (GCPS), including one pair of monozygotic twin boys with a de novo microdeletion involving the chromosomal band 7p13, where various clinical manifestations, in addition to GCPS, were recognized. Besides the twin pair, all patients are unrelated. Since there is a considerable lack of well-defined clinical delineation of the few patients with microdeletions involving 7p13 with GCPS described so far, we focus on the symptoms that are not typically related to GCPS, such as moderate psychomotor retardation, seizures, muscle fiber anomalies, cardiac anomalies, hyperglycemia, and hirsutism. Our observations suggest that in all cases of atypical GCPS, the presence of a cytogenetically detectable microdeletion or a submicroscopic deletion of 7p13 should be suspected. PMID- 11484202 TI - Unique family with Townes-Brocks syndrome, SALL1 mutation, and cardiac defects. AB - Townes-Brocks syndrome (TBS) is a condition with imperforate anus, hand anomalies, and ear malformations with sensorineural hearing loss. Many cases are sporadic. Within and between families, the phenotype displays striking variability. Recently, the disease-causing gene for TBS was identified as SALL1, a zinc finger transcription factor. Here, we report a three-generation family with seven affected individuals who have a novel SALL1 mutation. Unique cardiac anomalies seen in this family include lethal truncus arteriosus in one patient and a lethal complicated defect, including pulmonary valve atresia, in a second patient. These severe cardiac anomalies have not previously been reported in a familial case of TBS. This family and a review of the literature indicate that cardiac evaluation is warranted in all individuals with this disorder. In addition, hypoplastic thumbs were seen in two individuals in this family and should, therefore, be considered a true feature of TBS. PMID- 11484203 TI - Semilobar holoprosencephaly, coronal craniosynostosis, and multiple congenital anomalies: a severe expression of the Genoa syndrome or a newly recognized syndrome? AB - We report on a female newborn with holoprosencephaly, craniosynostosis, and multiple congenital anomalies including cloverleaf skull, Dandy-Walker malformation, bilateral microphthalmia, cleft soft palate, congenital scoliosis, hypoplastic nails and coarctation of aorta. Some of these features are consistent with the diagnosis of the Genoa syndrome, (MIM 601370) a rare autosomal recessive disorder recently described. The findings of other serious and previously undescribed malformations, however, raises the possibility of a newly recognized disorder. PMID- 11484204 TI - Deletions at chromosome regions 7q11.23 and 7q36 in a patient with Williams syndrome. AB - We report on a patient with Williams syndrome and a complex de novo chromosome rearrangement, including microdeletions at 7q11.23 and 7q36 and additional chromosomal material at 7q36. The nature of this additional material was elucidated by spectral karyotyping and first assigned to chromosome 22. Subsequent fluorescence in situ hybridization (FISH) experiments showed that it consisted of satellite material only. Refinement of the 7q36 breakpoint was performed with several FISH probes, showing a deletion distal to the triphalangeal thumb (TPT) region. The phenotype of the patient principally results from the microdeletion of the 7q11.23; the small deletion at 7qter and the extra satellite material may not be of clinical significance. PMID- 11484205 TI - Molecular cytogenetic characterization of a derivative chromosome 8 with an inverted duplication of 8p21.3-->p23.3 and a rearranged duplication of 8q24.13- >qter. AB - A derivative chromosome 8 was observed in a newborn boy who presented with low birth weight, multiple congenital anomalies, and dysmorphic face. The der(8) was further characterized at age 18 months by a high resolution G-banding analysis, spectral karyotyping, and fluorescence in situ hybridization (FISH) with multiple DNA probes. The karyotype was described as 46,XY,der(8)(qter-->q24.13::p21.3- >p23.3::p23.3-->qter), representing an inverted duplication of region 8p21.3- >p23.3 and a duplication of region 8q24.13-->qter, which attaches to the duplicated short arm segment at 8p21.3. Different from previously reported patients with an inverted duplication (8p), no deletion was detected in the distal region of 8p in this case. This young child had manifested a broad nasal bridge, micrognathia, cleft lip, hydrocephalus, partial agenesis of the corpus callosum, Dandy-Walker malformation, congenital heart defects, dysplastic kidneys, hydronephrosis, marked hypotonia, and significant psychomotor retardation. These features are compared with those commonly seen in cases with an inverted duplication of 8p and cases with a partial trisomy of 8q. PMID- 11484206 TI - Common MEFV mutations among Jewish ethnic groups in Israel: high frequency of carrier and phenotype III states and absence of a perceptible biological advantage for the carrier state. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disease, characterized by recurrent attacks of fever and inflammation of serosal membranes and gradual development of nephropathic amyloidosis. The recent cloning of the FMF gene (MEFV) and identification of disease-associated mutations in most patients made the direct determination of FMF carrier frequency feasible. The aim of the present study was to investigate the carrier rate of the most common MEFV mutations among different Jewish ethnic groups in Israel. Further, an attempt was made to elucidate the possible biological advantage that the heterozygote state may confer. Three hundred Ashkenazi, 101 Iraqi, and 120 Moroccan Jews were screened for the E148Q, V726A, and M694V mutations (at least two most common mutations per group), with a resulting overall carrier frequency in the respective ethnic group of 14%, 29%, and 21%. No difference in morbidity between Ashkenazi carriers and non-carriers of MEFV mutations was discerned, although an excess of febrile episodes in carriers of the V726A and in carriers of either V726A or E148Q was evident (P < 0.02 and P < 0.05, respectively). The frequency of subjects with two MEFV mutations but not expressing FMF (phenotype III) was 1:300 in Ashkenazi Jews and 1:25 in Iraqi Jews, exceeding the reported rate of overt FMF in these ethnic groups by 40-240 fold. These results affirm the high carrier rate among the studied Jewish ethnic groups in Israel and suggest that most subjects with FMF mutations are unaffected. PMID- 11484207 TI - XV-2c/KM-19 haplotype analysis of cystic fibrosis mutations in Mexican patients. AB - We analyzed 97 unrelated Mexican cystic fibrosis (CF) patients and their first degree relatives to study the association of XV2C/TaqI/KM19/PstI haplotypes with CF mutations in this population. Haplotype phases could be established in 148 CF and 110 normal chromosomes, and haplotype distributions of normal and CF chromosomes differed significantly (P < 0.001). DeltaF508 and G542X mutations accounted for 56% of CF chromosomes and were found to be associated with haplotype B in 97.2% and 72.7% of chromosomes, respectively. The haplotype distribution of CF chromosomes carrying other rare and unknown mutations was similar to that of normal chromosomes (P > 0.05), haplotypes A and C being the most frequent. This is in accordance with the extensive heterogeneity and the spectrum of mutations reported in Mexican CF patients. We also report the haplotype distribution of all informative chromosomes bearing rare mutations; some were found to be associated with previously reported haplotypes, whereas others were found on different haplotypes. Recombination or recurrence of mutations may explain these different associations, although other intragenic markers must be used to better understand the origin and dispersion of CF mutations in our country. XK haplotype analysis allowed carrier detection among sibs in 24.3% of families, showing that this method may be useful for carrier detection in populations with high allelic heterogeneity. PMID- 11484208 TI - Novel SNP at the common primer site of exon IIIa of FGFR2 gene causes error in molecular diagnosis of craniosynostosis syndrome. AB - Most mutations in Crouzon, Pfeiffer, and Apert syndromes are in the extracellular, third immunoglobulin-like domain and adjacent linker regions (exons IIIa and IIIc) of the fibroblast growth factor receptor 2 (FGFR2) gene. Using the published primers for PCR, a patient with Crouzon syndrome was found to be homozygous for a mutation that results in a Q289P amino acid substitution in FGFR2. Two additional patients; one with Apert syndrome and P253R mutation, the other with Pfeiffer syndrome and S267P mutation, also appeared to be homozygous. Using a new primer located 146 bp 5' of exon IIIa for PCR followed by sequencing revealed an A to G polymorphism at -62 [corrected] position of exon IIIa. All three patients were heterozygous for both the mutation and the polymorphism. These results indicate that the polymorphism and the mutation are not on the same chromosome. The single nucleotide polymorphism is located at the second to the last base of the 3' end of the published primer. This primer mismatch caused the failure of amplification of the normal chromosome and thus, the apparent homozygosity. The frequency of this novel polymorphism was determined to be 0.03 by studying 326 chromosomes from the general population. We propose that a new primer should be used for mutational analysis of exon IIIa of FGFR2 to avoid misdiagnosis caused by primer mismatch. PMID- 11484210 TI - Megacystis-microcolon-intestinal hypoperistalsis syndrome and aganglionosis in trisomy 18. AB - Ultrasonography at 23 weeks of gestation documented the presence of megacystis with horseshoe kidney, microcolon, intestinal malrotation, and decreased amniotic fluid volume. After pregnancy termination, an autopsy was performed. The external phenotype was diagnostic of the trisomy 18 syndrome confirmed by chromosome examination. The fetus also had a massively distended bladder with parchment-thin wall, microcolon, intestinal malrotation but no urethral obstruction or hydronephrosis. No ganglion cells were present in the colon or bladder. This has not been mentioned in other reported cases and, therefore, suggests pathogenic heterogeneity. The megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS) is a rare autosomal recessive condition of unknown pathogenesis whose genes map to 15q24. Thus, its previously undescribed presence in trisomy 18 further suggests etiologic heterogeneity. PMID- 11484209 TI - Kabuki syndrome-like features associated with a small ring chromosome X and XIST gene expression. AB - Although clinical features in Kabuki syndrome (KS; Niikawa-Kuroki syndrome) have been well defined, the underlying genetic mechanism still remains unclear. We report a 9-year-old girl with typical KS-like facial appearance, skeletal and dermatoglyphic abnormalities, severe mental retardation, and growth deficiency. In 60 of 100 GTG-banded metaphases from peripheral blood lymphocytes, a ring chromosome smaller than a G group chromosome was found, which, according to reverse painting, consisted of Xq11.1q13. The proband's karyotype was described as mos45,X/46,X,+r(X). Several loci were analyzed with fluorescence in situ hybridization (FISH) and microsatellite markers revealing that one r(X) breakpoint mapped proximal to DXS422 (Xp11.21) and the second mapped distal to XIST gene, between loci DXS128E and DXS441 (Xq13.2). Uniparental disomy for X and r(X) was excluded and the paternal origin of r(X) was identified. XIST expression was demonstrated by nested reverse transcription polymerase chain reaction (RT PCR) using primers spanning exons 5, 6i, and 6 in RNA prepared from lymphocytes. The observation of XIST expression is in contrast to two other cases in which the XIST gene was either not present on r(X) or not expressed. To our knowledge, this is the first case of Kabuki-like syndrome manifestations with r(X) and XIST expression. PMID- 11484211 TI - Infant with congenital erosions of the skin of several fingers and gastroschisis. PMID- 11484212 TI - Partial trisomy of 2p and neuroblastoma. PMID- 11484214 TI - Diagnosis of FS should not be made until PKS is ruled out. PMID- 11484218 TI - What are the dielectric "constants" of proteins and how to validate electrostatic models? AB - Implicit models for evaluation of electrostatic energies in proteins include dielectric constants that represent effect of the protein environment. Unfortunately, the results obtained by such models are very sensitive to the value used for the dielectric constant. Furthermore, the factors that determine the optimal value of these constants are far from being obvious. This review considers the meaning of the protein dielectric constants and the ways to determine their optimal values. It is pointed out that typical benchmarks for validation of electrostatic models cannot discriminate between consistent and inconsistent models. In particular, the observed pK(a) values of surface groups can be reproduced correctly by models with entirely incorrect physical features. Thus, we introduce a discriminative benchmark that only includes residues whose pK(a) values are shifted significantly from their values in water. We also use the semimacroscopic version of the protein dipole Langevin dipole (PDLD/S) formulation to generate a series of models that move gradually from microscopic to fully macroscopic models. These include the linear response version of the PDLD/S models, Poisson Boltzmann (PB)-type models, and Tanford Kirkwwod (TK)-type models. Using our different models and the discriminative benchmark, we show that the protein dielectric constant, epsilon(p), is not a universal constant but simply a parameter that depends on the model used. It is also shown in agreement with our previous works that epsilon(p) represents the factors that are not considered explicitly. The use of a discriminative benchmark appears to help not only in identifying nonphysical models but also in analyzing effects that are not reproduced in an accurate way by consistent models. These include the effect of water penetration and the effect of the protein reorganization. Finally, we show that the optimal dielectric constant for self-energies is not the optimal constant for charge-charge interactions. PMID- 11484219 TI - Structured disorder and conformational selection. AB - Traditionally, molecular disorder has been viewed as local or global instability. Molecules or regions displaying disorder have been considered inherently unstructured. The term has been routinely applied to cases for which no atomic coordinates can be derived from crystallized molecules. Yet, even when it appears that the molecules are disordered, prevailing conformations exist, with population times higher than those of all alternate conformations. Disordered molecules are the outcome of rugged energy landscapes away from the native state around the bottom of the funnel. Ruggedness has a biological function, creating a distribution of structured conformers that bind via conformational selection, driving association and multimolecular complex formation, whether chain-linked in folding or unlinked in binding. We classify disordered molecules into two types. The first type possesses a hydrophobic core. Here, even if the native conformation is unstable, it still has a large enough population time, enabling its experimental detection. In the second type, no such hydrophobic core exists. Hence, the native conformations of molecules belonging to this category have shorter population times, hindering their experimental detection. Although there is a continuum of distribution of hydrophobic cores in proteins, an empirical, statistically based hydrophobicity function may be used as a guideline for distinguishing the two disordered molecule types. Furthermore, the two types relate to steps in the protein folding reaction. With respect to protein design, this leads us to propose that engineering-optimized specific electrostatic interactions to avoid electrostatic repulsion would reduce the type I disordered state, driving the molten globule (MG) --> native (N) state. In contrast, for overcoming the type II disordered state, in addition to specific interactions, a stronger hydrophobic core is also indicated, leading to the denatured --> MG --> N state. PMID- 11484220 TI - Structure of bovine alpha-1,3-galactosyltransferase and its complexes with UDP and DPGal inferred from molecular modeling. AB - A homology model of alpha-1,3-galactosyltransferase (alpha-1,3-GalT), the retaining enzyme responsible for the formation of alpha-galactosyl epitopes, has been developed by means of molecular modeling using the SpsA glycosyltransferase structure. A protein-ligand docking approach was used to model alpha-1,3-GalT complexed with UDP and UDP-Gal. The comparison of structural features found in the alpha-1,3-GalT homology model with available structural data on this class of enzymes revealed similarities in the UDP-binding pocket. In the predicted structure of the complexes, the pyrophosphate interacts with the DVD motif (Asp 225, Val-226, and Asp-227) of alpha-1,3-GalT through the Mn(2+) cation. The uridine part of the UDP binds into the well-defined cavity that consists of Phe 134, Tyr-139, Ile-140, Val-136, Arg-194, Arg-202, Lys-209, Asp-173, His-218, and Thr-137 in a conformation that is generally observed in the crystal structures of other glycosyltransferase complexes. PMID- 11484221 TI - Computational study of lipid-destabilizing protein fragments: towards a comprehensive view of tilted peptides. AB - Tilted peptides are short sequence fragments (10-20 residues long) that possess an asymmetric hydrophobicity gradient along their sequence when they are helical. Due to this gradient, they adopt a tilted orientation towards a single lipid/water interface and destabilize the lipids. We have detected those peptides in many different proteins with various functions. While being all tilted oriented at a single lipid/water interface, no consensus sequence can be evidenced. In order to better understand the relationships between their lipid destabilizing activity and their properties, we used IMPALA to classify the tilted peptides. This method allows the study of interactions between a peptide and a modeled lipid bilayer using simple restraint functions designed to mimic some of the membrane properties. We predict that tilted peptides have access to a wide conformational space in membranes, in contrast to transmembrane and amphipathic helices. In agreement with previous studies, we suggest that those metastable configurations could lead to the perturbation of the acyl chains organization and could be a general mechanism for lipid destabilization. Our results further suggest that tilted peptides fall into two classes: those from proteins acting on membrane behave differently than destabilizing fragments from interfacial proteins. While the former have equal access to the two layers of the membrane, the latter are confined within a single lipid layer. This could be in relation with the organization of lipid substrate on which the peptides physiologically act. PMID- 11484222 TI - Modeling of the metallo-beta-lactamase from B. fragilis: structural and dynamic effects of inhibitor binding. AB - The structure and dynamics of an inhibitor-bound complex of the metallo-beta lactamase from Bacteroides fragilis are studied by using molecular dynamics. A search of the conformational space was performed to obtain three distinct models of the complex, which were then subjected to solvated molecular dynamics. A solvated molecular dynamics study of the apo protein was performed to serve as a baseline for comparison with the bound simulations. We find loop conformation changes due to binding as well as a decrease in flexibility of the protein as a whole and especially in the major loop of the beta-lactamase. We report the structural and dynamical features of the inhibitor-bound and apo models, as well as experimentally measurable quantities, which should be capable of distinguishing the two binding modes we have determined. PMID- 11484223 TI - Molecular dynamics simulations of the conformational changes of the glutamate receptor ligand-binding core in the presence of glutamate and kainate. AB - Excitatory synaptic transmission is mediated by ionotropic glutamate receptors (iGluRs) through the induced transient opening of transmembrane ion channels. The three-dimensional structure of the extracellular ligand-binding core of iGluRs shares the overall features of bacterial periplasmic binding proteins (PBPs). In both families of proteins, the ligand-binding site is arranged in two domains separated by a cleft and connected by two peptide stretches. PBPs undergo a typical hinge motion of the two domains associated with ligand binding that leads to a conformational change from an open to a closed form. The common architecture suggests a similar closing mechanism in the ligand-binding core of iGluRs induced by the binding of specific agonists. Starting from the experimentally determined kainate-bound closed form of the S1S2 GluR2 construct, we have studied by means of molecular dynamics simulations the opening motion of the ligand-binding core in the presence and in the absence of both glutamate and kainate. Our results suggest that the opening/closing interdomain hinge motions are coupled to conformational changes in the insertion region of the transmembrane segments. These changes are triggered by the interaction of the agonists with the essential Glu 209 residue. A plausible mechanism for the coupling of agonist binding to channel gating is discussed. PMID- 11484224 TI - Structure of a legume lectin from the bark of Robinia pseudoacacia and its complex with N-acetylgalactosamine. AB - The structure of the bark lectin RPbAI (isoform A4) from Robinia pseudoacacia has been determined by protein crystallography both in the free form and complexed with N-acetylgalactosamine. The free form is refined at 1.80 A resolution to an R factor of 18.9% whereas the complexed structure has an R-factor of 19.7% at 2.05 A resolution. Both structures are compared to each other and to other available legume lectin structures. The polypeptide chains of the two structures exhibit the characteristic legume lectin tertiary fold. The quaternary structure resembles that of the Phaseolus vulgaris lectin, the soybean agglutinin, and the Dolichos biflorus lectin, but displays some unique features leading to the extreme stability of this lectin. PMID- 11484225 TI - A conserved helix-unfolding motif in the naturally unfolded proteins. AB - Among the naturally unfolded proteins there are many polypeptides that retain an extended conformation in the absence of any apparent signal. Using sequence alignment and secondary structure prediction tools, a conserved (LS/SL)(D/E)(D/E)(D/E)X(E/D) motif is uncovered in the vicinity of the N-terminus of their unfolded helices. A comparison of these data with published observations allows one to propose that the (LS/SL)(D/E)(D/E)(D/E)X(E/D) motif is a helix unfolding signal. Furthermore, the strong similarity between this motif and the STXXDE casein kinase II phosphorylation site suggests a regulatory mechanism for the naturally unfolded proteins within the cell. PMID- 11484226 TI - Quantum mechanics simulation of protein dynamics on long timescale. AB - Protein structure and dynamics are the keys to a wide range of problems in biology. In principle, both can be fully understood by using quantum mechanics as the ultimate tool to unveil the molecular interactions involved. Indeed, quantum mechanics of atoms and molecules have come to play a central role in chemistry and physics. In practice, however, direct application of quantum mechanics to protein systems has been prohibited by the large molecular size of proteins. As a consequence, there is no general quantum mechanical treatment that not only exceeds the accuracy of state-of-the-art empirical models for proteins but also maintains the efficiency needed for extensive sampling in the conformational space, a requirement mandated by the complexity of protein systems. Here we show that, given recent developments in methods, a general quantum mechanical-based treatment can be constructed. We report a molecular dynamics simulation of a protein, crambin, in solution for 350 ps in which we combine a semiempirical quantum-mechanical description of the entire protein with a description of the surrounding solvent, and solvent-protein interactions based on a molecular mechanics force field. Comparison with a recent very high-resolution crystal structure of crambin (Jelsch et al., Proc Natl Acad Sci USA 2000;102:2246-2251) shows that geometrical detail is better reproduced in this simulation than when several alternate molecular mechanics force fields are used to describe the entire system of protein and solvent, even though the structure is no less flexible. Individual atomic charges deviate in both directions from "canonical" values, and some charge transfer is found between the N and C-termini. The capability of simulating protein dynamics on and beyond the few hundred ps timescale with a demonstrably accurate quantum mechanical model will bring new opportunities to extend our understanding of a range of basic processes in biology such as molecular recognition and enzyme catalysis. PMID- 11484227 TI - Interactions of Streptomyces griseus aminopeptidase with amino acid reaction products and their implications toward a catalytic mechanism. AB - Streptomyces griseus aminopeptidase (SGAP) is a double-zinc exopeptidase with a high preference toward large hydrophobic amino-terminus residues. It is a monomer of a relatively low molecular weight (30 kDa), it is heat stable, it displays a high and efficient catalytic turnover, and its activity is modulated by calcium ions. The small size, high activity, and heat stability make SGAP a very attractive enzyme for various biotechnological applications, among which is the processing of recombinant DNA proteins and fusion protein products. Several free amino acids, such as phenylalanine, leucine, and methionine, were found to act as weak inhibitors of SGAP and hence were chosen for structural studies. These inhibitors can potentially be regarded as product analogs because one of the products obtained in a normal enzymatic reaction is the cleaved amino terminal amino acid of the substrate. The current study includes the X-ray crystallographic analysis of the SGAP complexes with methionine (1.53 A resolution), leucine (1.70 A resolution), and phenylalanine (1.80 A resolution). These three high-resolution structures have been used to fully characterize the SGAP active site and to identify some of the functional groups of the enzyme that are involved in enzyme-substrate and enzyme-product interactions. A unique binding site for the terminal amine group of the substrate (including the side chains of Glu131 and Asp160, as well as the carbonyl group of Arg202) is indicated to play an important role in the binding and orientation of both the substrate and the product of the catalytic reaction. These studies also suggest that Glu131 and Tyr246 are directly involved in the catalytic mechanism of the enzyme. Both of these residues seem to be important for substrate binding and orientation, as well as the stabilization of the tetrahedral transition state of the enzyme-substrate complex. Glu131 is specifically suggested to function as a general base during catalysis by promoting the nucleophilic attack of the zinc bound water/hydroxide on the substrate carbonyl carbon. The structures of the three SGAP complexes are compared with recent structures of three related aminopeptidases: Aeromonas proteolytica aminopeptidase (AAP), leucine aminopeptidase (LAP), and methionine aminopeptidase (MAP) and their complexes with corresponding inhibitors and analogs. These structural results have been used for the simulation of several species along the reaction coordinate and for the suggestion of a general scheme for the proteolytic reaction catalyzed by SGAP. PMID- 11484228 TI - One small step on privacy. HHS action addresses some provider concerns. PMID- 11484229 TI - Peaches and cream. Despite Medicare changes, rural hospital program likely to remain untouched. PMID- 11484230 TI - Another settlement for CHW. Troubled Sacramento, Calif., division pays $2.9 million in cost-report case. PMID- 11484231 TI - Hospital closure pace still high. PMID- 11484232 TI - Antikickback law yields results. Hundreds of docs may pay up in lab probe; criminal charges a possibility. PMID- 11484233 TI - Technology's Rip Van Winkles. Hospitals are waking up, slowly, to the need to embrace computers and automation. PMID- 11484234 TI - Tax-exempt bids get complicated. PMID- 11484235 TI - Raising awareness or ire? Insurer's ad campaign targets medical errors, prompts provider indignation. PMID- 11484236 TI - Where the money goes. Two big insurers try different tacks on patient safety. PMID- 11484237 TI - Patient dumping cases shoot up. PMID- 11484238 TI - Unrest amid the reformation. Bush offers eight principles to reform Medicare; hospital execs want details. PMID- 11484239 TI - Take a meeting. Drug firms hosting many more events for docs. PMID- 11484240 TI - [Omentoplasty in surgical management of postpulmonectomy pleural empyema]. AB - A method of treating postpulmonectomy pleural empyema, practically implemented in this country for the first time, is described. Three patients are subjected to operation. Right pulmonectomy for lung cancer and chronic inflammatory process is done twice, and left pleuropulmonectomy for tuberculosis--once. Postpulmonectomy empyema persists in all three cases regardless of the adequate continuous suction drainage and intrapleural antiseptic management. Anaerobic and gram-negative flora is isolated. The size and location of the intrapleural cavity being cured are assayed by CT, thoracic ultrasonography and fistulography. In none of the patients is clinical and FBS evidence of bronchopleural fistula established. The operative procedure consists in resection of a 4 cm segment from the underlying rib in the drainage zone, and further cavity treatment under thorascopic control. Minor median laparotomy and skeletization of the greater omentum are performed preserving a major nutrient vessel depending on the location of the cavity. The omentum is drived into the pleural cavity through a parietal opening of the diaphragm, measuring 3-4 fingerbreadths. Pleural cavity drainage is carried out according to Redon. Two patients run an uneventful postoperative course. One female patient develops pylorospasm successfully cured by spasmolytic therapy and H2 blockers. CT and thoracic echography do not show presence of residual cavities. The patients are free of any complaints over periods ranging from 6 to 16 months postoperatively. The inference is reached that omentoplasty is a new method promoting successful elimination of both postpulmonectomy empyema, and other residual pleural cavities as well, with or without bronchopleural fistula. PMID- 11484241 TI - [Continent derivation of urine (a Modo) Mainz II]. AB - The improved intestinal dissection technique and antibiotic agents, and the development of new resorptive suture materials have led to renewed surge of interest in ureterosigmoidostomy as a practicable procedure for continent derivation. Based on data from urodynamic assessment of diverse forms of urinary derivation, M Fisch and R Hohenfellner in 1991 propose a modification of the classical technique of ureterosigmoidostomy, coined with the term Sigmarectum pouch or Mainz pouch (M Fisch, R. Wammack, R Hohenfellner 1991). Over the period 1992 through 1999, 46 operations type "Mainz II" are performed in the Department of Urology--University Hospital "Alexandrovska". The series includes 41 men and 5 women with age range 42 to 80 years. All patients present urinary bladder tumors. The usual surgical technique is used. The severest postoperative complications include: urinary fistulae--3 (6.52%), local relapse--4 (8.69%) and distant metastases--2 (4.35%). In six patients the outcome is fatal (0.7%). This type of continent derivation following cystectomy is recommended since it is convenient, practically atraumatic and well tolerated by the patients. Complications encountered seldom necessitate operative intervention. PMID- 11484242 TI - [Kidney diseases most often considered as indications for nephrectomy]. AB - Nephrectomy is a radical operation successfully used over more than a century. It should be resorted to only in exceptional situations whenever an organ salvaging operation is precarious for the patient's health (T Patrashkov, 1980). The indications for nephrectomy depend on the type of disease, extent of renal damage, state of the second kidney and the patient's general condition, established by the basic examination methods in urology (T Patrashkov 1982). The study covers 388 nephrectomies in cases presenting diverse diseases of the kidney and ureter, diagnosed and treated in the Department of Urology--University Hospital "Alexandrovska" in the period 1990 to 1995. The commonest causes leading to nephrectomy comprise: 1. Neoplasms of kidney and ureter--134 (34.54%). 1.1. Parenchymal tumors--116 (29.90%)/ 1.2. Papillary tumors--18 (4.64%). 2. Pyonephrosis--88 (22.68%). 3. Nephrolithiasis (presence of renal calculi)--53 (13.66%). 4. Secondary operations of the kidney and ureter--46 (11.86%). 5. Hydronephrosis--38 (9.80%). 6. Anomalies (hypoplasia)--8 (2.06%). 7. Cystic diseases--7 (1.80%). 8. Tuberculosis of kidney--6 (1.55%). 9. Renovasal hypertension--4 (1.02%). 10. Nephrectomy for other diseases--4 (1.02%). As shown by the results the rate of nephrectomy undertaken for renal malignancy is still the highest which is by no means considered as a favourable diagnostic sign. PMID- 11484243 TI - [Microbiological study of material from upper airways and urogenital tract as a condition for effective treatment in urology]. PMID- 11484244 TI - [Tumors of the appendix]. AB - Neoplasms involving the vermiform outgrowth are among the rarely met with conditions, accounting for 0.1 to 0.5 per cent of all gastrointestinal tract tumors. This is a report on personal observation of three patients presenting tumors of the appendix, diagnosed over the period 1987 to 1995. As shown by the results, tumors of the appendix run a clinical course characterized by two distinct forms: clinical picture of acute appendicitis--two cases, and clinical picture of tumor of the colon--one case. It is underscored that intraoperative diagnosis is extremely difficult. The diagnosis is usually made by pathoanatomist, while the therapeutic approach and operative tactics are determined on the ground of histomorphological study results and local operative status. The scope of operative intervention includes appendectomy and right hemicolectomy. PMID- 11484245 TI - [Endorectal echography--diagnostic relevance in pararectal and perivesical region diseases]. AB - The diagnostic relevance of endorectal ultrasonography is assessed in a series of 56 patients presenting surgical, urological and obstetric-gynecological conditions, distributed as follows: carcinoma of the rectum (30 cases), prostate gland pathology (12 men) and uterine neck carcinoma (14 women). Endorectal echography contributes greatly to preoperative staging of the patients with rectal carcinoma. In prostate gland diseases it proves helpful in making the diagnosis before and after the treatment. In conclusion it is underscored that endorectal echography is a noninvasive procedure highly effective economically. PMID- 11484246 TI - [Tactics and operative methods in treating complicated colorectal cancer]. AB - Over the period 1993-1998, a total of 385 patients with complicated colorectal cancer are subjected to treatment in the clinic of emergency surgery--University Hospital "Queen Giovanna", Sofia. Obturation is the commonest form of complication--43.8 per cent, perforation within the tumor or diastasis noted in 27 cases (7.01%), paratumor abscesses and infiltrates--in 8.3 per cent and rectohemorrhage--in 4.2 per cent. The therapeutic and operative approach, and the scope of surgery are dependent on a multitude of factors which should be given due consideration by the surgeon. In each patient presenting complicated colorectal carcinoma it is mandatory to make a precise and individual choice of the extent of operative intervention, consistent with the patient's general condition and contributing to eliminate the life-endangering underlying cause. PMID- 11484247 TI - [Retinoic acid--a teratogenic factor in caudal dysgenesis. Experimental models and hypothesis for pathogenesis]. AB - Using teratogen--trans-retinoic acid (Sigma Chemical)--caudal malformations are experimentally induced in mice of the Spriche-Dolly breed. For the purpose the aforementioned reagent is injected into pregnant mice at 6-hour intervals dose 29 mg/kg. In stages between two and four paired somites, supravital staining with Nile blue sulfite (in 1:50,000 Ringer solution at 37 degrees C for 30 min) is done, followed by histological assessment of tissue sections. The malformation noted are summed up in an attempt to draw conclusions about their presumable pathogenesis. PMID- 11484248 TI - [Trauma to the major upper airways]. PMID- 11484249 TI - [Operative treatment of necrotic pancreatitis]. AB - Treatment of necrotic pancreatitis patients is still disputable, posing serious problems and a real challenge to surgeons. The surgeon's approach to the problem ranges from strictly conservative to extremely aggressive. In this paper are discussed the indications, timing and operative technique used in the treatment of necrotic pancreatitis (NP), with special attention focused on the method suggested by the authors--necrotomy and closed continuous lavage of the retroperitoneum. An algorithm of the therapeutic approach to patients presenting acute pancreatitis is likewise presented. All patients with necrotic pancreatitis are routinely referred for treatment in the intensive care unit (ICU), and mandatorily subjected to antibiotherapy. Only one-third of the cases developing infected pancreatic necrosis (IPN) within 3 weeks of admission undergo operation. Lethality among those operated on is reduced to 13 per cent, and among NP patients--4 per cent. The new concept for therapeutic approach to necrotic pancreatitis patients according to which only cases presenting evidence of infected pancreatic necrosis are liable to surgery, whereas those with SPN should be treated conservatively, is strongly supported. PMID- 11484250 TI - [A rare case of extrapleural hematoma]. PMID- 11484251 TI - [Plastic and reconstructive surgery of the bronchial tree]. AB - Bronchoplastic and reconstructive operations (BPRO) are a major issue in the broad methodological spectrum of thoracic surgery. It is the aim of the study to analyze the indications, operative technique and results of such operations on the basis of experience gained in the Clinic of Thoracic Surgery over a 5-year period. A total of 19 patients (14 men and 5 women) at mean age 50.7 y (range 16 to 70 y) are operated. By histological variant of the tumor operated on, the patients are distributed as follows: carcinoid--4 cases, fibromas--1, squamous cell carcinoma--10, adenocarcinoma--1, bronchoalveolar carcinoma--1, small-cell carcinoma--1 and leiomyosarcoma--one. The reconstructive operations performed include: isolated bronchus resection--2, right upper lobectomy with cuff resection--7, right upper bilobectomy with cuff resection--2, left upper lobectomy with cuff resection--7 (in two instances in conjunction with angioplasty), and left lower lobectomy with cuff resection and angioplasty--one. No intraoperative and perioperative lethality (within 30 days) is recorded. An overweight female patient with diabetes hardly lending itself to compensation develops severe suppuration. In two instances serious concurrent complications necessitate reoperation. Overall postoperative hospital stay--20 days; without the 3 severe complications--12.8 days. One patient dies of brain metastases within 6 months of the intervention. The survivorship term in the remainder varies from 1 year to 4 years 9 months, averaging 31 months. There are no stenoses or granulations of the anastomoses requiring endoscopic treatment. Presumably, BPRO are an adequate therapeutic approach to patients presenting centrally located malignant and benign tumors. The results of their application in the series being examined are estimated as very good. PMID- 11484252 TI - [Gelatinoform carcinoma of the cecum--a case report]. PMID- 11484253 TI - [A rare case of an adult male with relapsed esophageal carcinoma with a successful operation]. PMID- 11484254 TI - [An original operative approach in esophageal rupture (a report of a case)]. PMID- 11484255 TI - [A variant of blood supply to the upper limb in man with a special reference to the hazards of erroneous intra-arterial administration of drugs]. PMID- 11484256 TI - [Combined variation of the a. subclavia and a. thyroidea superior branches]. PMID- 11484257 TI - [Surgical tactics and surgical techniques in echinococcal cysts in the lung in children]. AB - The differentiated surgical approach to hydatid lung cysts and the operative techniques used in 343 children treated in the Department of Pediatric Surgery over 24 years (1975-1998) are presented. The left lung is involved in 143 children (41.7%), the right--in 155 (45.2%), whereas in 45 cases (13.1%) it is a matter of bilateral location. Infectious complications occur in 198 cases (57.7%). In compliance with the anatomical location, size, extent and accompanying complications two surgical procedures are mainly used: 1) Total removal of the cyst without disrupting its continuity (closed echinococcotomy) in 19 per cent, and 2) Evacuation of the cyst fluid, as well as the germinative membrane, followed by appropriate management of the residual fibrous cavity in 81 per cent of cysts. Echinococcotomy with suture obliteration of the residual cavity in conjunction with sparing lung resection, yielding superior functional results postoperatively, is the method of choice. PMID- 11484258 TI - [Preoperative staging on liver metastasis from colorectal carcinoma]. AB - Preoperative staging of metastatic liver cancer (MLC) from colorectal carcinoma (CRC) is conducted with the purpose to identify potentially resectable MLC cases. Fifty-two patients undergo laparoscopic study (4.1% all endoscopies), and three- laparoscopic ultrasonography. Laparoscope E Wolf and 7.5 MHz linear transducer obtained from the Aloka Company are employed. The study covers 24 women and 28 men with mean age 61.6 years (range 23-87 y). MLC is diagnosed in 35.1% (13/37) of colonic, and in 73.3% (11/15) of rectal carcinoma cases. Furthermore, 69.3% of the patients (36/52) are in M-stage, accounted for by peritoneal or hepatic metastases. The size of lesions measures less than 2 cm in 25.0%, and less than 1 cm in 12.5% of the cases. Solitary metastasis is recorded in 8.3 per cent. In the event of isolated involvement of the left lobe the location of MLC is in the region of II-III segment. In one patient a single metastasis in VII segment, measuring 0.5/1.0 cm, missed by the imaging methods and laparoscopy, is documented by laparoscopic ultrasonography. Echographic diagnosis proves inaccurate in 16.7%, and false negative--in 12.5 per cent. In further three patients (12.5%) the conclusion reached is uncertain, with primary liver cancer being also considered. In 23.1% MLC from colonic carcinoma is situated entirely intrahepatally, and is not accessible to endoscopic inspection without laparoscopic and/or intraoperative ultrasonography; the latter is indicated in all T3 and T4 patients. Potential resectability is established in 16.7% of cases with liver metastases from CRC. PMID- 11484259 TI - [Risk of drug-induced acute pancreatitis following propofol anesthesia used in abdominal operations]. AB - The postoperative pancreatitis is a well-known complication. More than eighty five drugs have been reported to have induced postoperative pancreatitis. Twenty five cases of Propofol-induced pancreatitis have been reported till now. The relation of pancreatitis to Propofol (Diprivan; Zeneca; London, UK) is considered as a possible one, but has been proved. The goal of the authors is to reveal the plasma lipid and amylase level dynamics in patients after Propofol anaesthesia. The clinicians have to consider this possible relation, because of the growing use of Propofol recently. PMID- 11484260 TI - [Post-gastrectomy reconstruction versus enteral alimentation--a one year follow up study]. AB - This is a report on various post-gastrectomy reconstructive procedures, proceeding from personal experience had with 14 patients undergoing gastrectomy for carcinoma of the stomach and five patients with operations of esophagus and stomach for various pathological conditions. An assessment is done of the reconstructive methods used on the ground of objective indicators. All patients are subjected to enteral alimentation through nutritive jejunostomy. In the pre- and postoperatived periods, the trophic state of patients is evaluated on basis of biochemical, anthropometric and immunologic study data. Postoperatively, in those with small-intestinal reservoir a lower degree negative symptomatology is established, along with life style improvement. Post-gastrectomy patients presenting I-II (UICC) stage of the oncological disease are indicated for small intestinal reservoir formation. In the overall therapeutic approach to post gastrectomy patients adequate clinical therapeutic feeding proves absolutely indispensable. PMID- 11484261 TI - [Echinococcosis of the spleen-notes on some diagnostic and therapeutic problems]. AB - Twenty patients with hydatid disease of the spleen--14 presenting isolated location within the spleen, five--combined with liver cyst, and one--polyorganic involvement--are operated in the Department of general and operative surgery- Sofia over the period January 1985 through December 1998. The currently used methods of echinococcosis diagnosing are described, and the indications for splenectomy and organ salvaging echinococcotomy in the aforementioned localization are discussed. PMID- 11484262 TI - [Residual cavity evacuation following echinococcotomy-methods and results]. AB - The results of operative management of liver hydatid disease are still estimated as unsatisfactory, with evacuation of the residual cavity after echinococcotomy a major problem faced. The study covers 100 patients, aged 15 to 75 years, presenting a total of 146 liver hydatid cysts, operated in the surgical diseases department of the Higher Medical institute--Pleven over the period 1992 through 1998. In 72 cases it is a matter of a solitary formation, and in the remainder (28) two or more cysts are present. A closed method is used in the operative management of 32 cysts, semiclosed (with or without autoplombage, capitonnage respectively)--in 98, and partial resection or pericystectomy--in sixteen. To monitor the residual cavity US imaging and/or CAT are employed. Surgical approaches and methods applied in the various locations, and the respective results obtained are discussed. Emphasis is laid on the safety afforded by the procedure including resection of the prominating fibrous wall (resection du dome saillant) and tight closure around the tubular drain. A maximum reduction of the residual cavity associated with the possibility of simultaneous evacuation of the exudate secreted reduce to a minimum the likelihood of persisting residual formation postoperatively. PMID- 11484263 TI - [Plastic surgical repair according to Lazar da Silva in large median eventrations]. AB - Lazaro da Silva's technique is postulated as a method free of relapses provided autologous endogenous tissues are made use of. It consists in the raising of three flaps cut from the hernial sac, anterior and posterior sheats of the rectus abdominis muscles. The suture lines lie in three different anatomical planes, thereby promoting restoration of the integrity and anatomical patterns of the anterior abdominal wall. The procedure is used in three patients presenting xiphopubic eventration of > 5 years longstanding. During the follow-up study (1.5 to 3 years) no relapses are registered. The technique described yields very encouraging results. Further comprehensive studies along this line will demonstrate whether or not the reputation of the procedure gained abroad will be confirmed. PMID- 11484264 TI - [Continent preternatural anus--histomorphological and functional aspects]. AB - Transplantation of a free smooth-muscular cuff around a terminal large-intestinal segment, brought out as preternatural anus, secures high-degree continence and controlled evacuatory rhythm of the intestinal content. The preserved structure of the muscular layers in the cuff wall and the myogenic nature of smooth muscle tone are preconditions for functional activity manifestation in conditions of denervation, isolation and even after block of neurons and intramural ganglia. Five preparations comprising the smooth muscle cuff and recipient large intestinal segment undergo histomorphologic and electromyographic study within a year of transplantation. In a series of 31 patients with drawn out continent preternatural anus the thickness of the abdominal wall together with the smooth muscle cuff graft is echographically registered at the end of the first postoperative year. The echographically documented abdominal wall thickness is compared with the histomorphologically established values of the indicator under study. Analysis of the functional activity of the smooth-muscle-cuff transplant is also done. PMID- 11484265 TI - [Iatrogenic lesions of hepatocholedocus in cholecystectomy]. AB - This is a report on nine patients presenting different intraoperative lesions to hepatocholedochus, some of them diagnosed in the course of operation, and others- after the intervention. A variety of plastic repairs of the hepatocholedochus, including simple suture, Saple's operation or hepatodigestive anastomosis after Longmayre are performed. The probability of inflicting lesions, the underlying causes and the therapeutic approach to concrete cases are analyzed. Abiding to the principles of surgery is strongly recommended, such as: sufficient and appropriate operative access, atraumatic manipulations, drainage of the gallbladder bed and the like, with a view to perclude serious and sometimes fatal complications' occurrence, as well as making early diagnosis and accordingly taking the most opportune surgical solution to the problem faced. PMID- 11484266 TI - [Late results from operative treatment of complicated chronic cholecystitis]. AB - Assessment is made of the long-term results in 75/232 patients with chronic cholecystitis, operated in the surgical clinic of the Military hospital--Plovdiv over the period 1991 through 1995. To analyze the questionnaire study results, the patients are divided up into three groups, as follows: Group one--61 cases (81/3%) where the complaints are completely eliminated. Group two--13 cases (17/1%) where the condition shows improvement, but during physical atress and nonobservance of the dietary regimen, heaviness and blunt pain in the right subcostal area occur. Group three--one patient with persisting complaints and lethal outcome. Evaluation of the long-term results shows that the positive effect attained is persistent and durable. Cholecystectomy is taken to be as the most rational and often used operative intervention. PMID- 11484267 TI - [Some etiologic factors in condom-induced allergic contact balanitis]. AB - Clinical cases presenting allergic contact balanitis from condoms are studied using a combination of patch testing and chemical analysis by gas chromatography and high-resolving liquid chromatography to identify the causative agents involved. It has been previously established that zinc ethylphenyl dithiocarbamate (ZEPC)--a dithiocarbamate-type accelerant (DTC)--is an etiological factor in the event of contact allergy to condoms. Later it is confirmed that DTCs, such as zinc dimethyl dithiocarbamate (ZDMC), zinc diethyl dithiocarbamate (ZDEC) and zinc dibutyl dithiocarbamate and amines, such as dimethylamine (DMA), diethylamine (DEA) and piperidine (PIP) are likewise implicated in condom-induced contact balanitis. As shown by the investigation along this line, although thiurams are believed to be more allergenic, as compared to the corresponding DTCs and amines, both DTCs (ZDMC, ZDEC, ZDBC, ZEPC) and amines (DMA, DEA and PIP) are noteworthy etiological factors of allergic contact balanitis from condoms. PMID- 11484268 TI - [Local recurrence after radical surgery for rectal carcinoma]. AB - The incidence rate of local recurrences after radical operative treatment of rectal carcinoma and the risk factors involved, methods of early diagnosis and surgical therapeutic approach are comprehensively analyzed. Reference is also made to 86 scientific publications on the issue. During the 20-year follow-up period, no tendency of the incidence of local rectal carcinoma relapses to decrease is noted. The risk factors contributing to their occurrence are distributed in two groups depending on the primary tumor characteristics (location, stage of development, locoregional spreading, differentiation degree, genetic features), and operative intervention used (type, distal resection line, application of total mesorectal excision, extensive minor-pelvis lymph dissection etc.). The early diagnosis is based on mandatorily performed procedures, such as rectoscopy (coloscopy), endo-ultrasonography with purposeful thin-needle biopsy, assessment of tumor markers and immunoscintigraphy. The surgical tactics in coping with a local recurrence of rectal carcinoma becomes increasingly aggressive and radical by resorting to re-resection, abdomino-perineal extirpation, en-bloc resections and even minor pelvic exenteration. Postoperative 5-year survivorship amounts to 54.5 per cent. PMID- 11484269 TI - [Antibiotic prophylaxis against serious wound infection in prosthetic arterial replacement]. AB - Wound infection in arterial prosthetic replacement is the most hazardous complication in vascular surgery, associated with a high incidence rate of invalidity and lethality. The choice of antibiotic and administration scheme play a major role in its prevention. Clinical experience with implantation of synthetic vascular prostheses in a series of 103 patients where Cefuroxim prophylaxis according to a standard scheme is used (described in detail) is comprehensively analyzed. Wound infection grading is done in compliance with Salzman's classification. Against the background of antibiotic prophylaxis deep wound infection with vascular prosthesis involvement is registered in 1.94 per cent of operated patients only which is consistent with literature data on the issue. The obtained results are estimated as very good, and justify Cefuroxim prophylaxis recommendation to the wide practice for maximal reduction of deep wound infection in prosthetic replacement of arteries. PMID- 11484270 TI - [Acute purulent peritonitis-classification and modern aspects]. PMID- 11484271 TI - [Bacteroides infection in oral surgery-a literature review and clinical cases]. PMID- 11484272 TI - [A case of infection of a dacron prosthesis of the subclavian artery, replaced by an autologous conduit from the iliac artery]. PMID- 11484273 TI - [A case of carcinoma of the cecum in a background of situs viscerum inversus totalis]. PMID- 11484274 TI - [A rare localization of extranodal malignant lymphoma (mammary gland)]. PMID- 11484275 TI - [Preoperative diagnosis of a rare combination of small intestinal invagination and Meckel's diverticulum]. PMID- 11484276 TI - [Pyoderma fistulans significa (PFS). A report of two cases]. PMID- 11484277 TI - [Widened right paramedial segmentectomy comprising IV hepatic segment in a patient with primary liver carcinoma]. AB - This is a report on a rare operative intervention undertaken for primary liver carcinoma, located in the central hepatic segments (after Couinaud). The operative technique, intraoperative carcinoma resectability and the tools of hemorrhage control are discussed. PMID- 11484278 TI - [Arterial reoperation of the aorto-femoral segment]. AB - Late rethromboses of aortofemoral prostheses following arterial reconstructions for aortoiliac occlusive disease (AIOD) are among the serious challenges faced by vascular surgeons. Hyperplasia of the intima, progressive atherosclerotic process, as well as regional hemodynamics impairment are taken to be the major factors promoting late rethrombosis development after arterial bypass for AIOD. It is the purpose of the study to assay the underlying causes and terms of rethrombosis in aortofemoral bypasses for AIOD, and analyze the results of the arterial reoperation procedures used. Over the period 1990-1997, 408 aortofemoral bypass procedures are performed in the University Hospital "St Catherine", with rethrombosis of the prostheses occurring in 19.45 per cent of the branches within 5 years of the initial intervention. Five methods of reoperation are used, with optimal results--100 per cent patency of prostheses--attained in aortobifemoral re-bypasses. Nevertheless, this particular operative procedure is considered as practicable in isolated, properly selected, relatively young patients free of associated diseases. In adults or patients with health problems preference is given to rather simple procedures, if possible performed under regional anesthesia, such as thrombectomy of the prosthesis (TE) using a deep femoral artery patch and/or sequence distal bypass towards arteria poplitea. The latter method accounts for limb salvage in 97.14 per cent of cases. A satisfactory outcome (88.58% preserved limbs) is likewise secured by resection of the failing distal anastomosis, and its substitution for neobifurcation towards the unobstructed AFS and APF. In elder and/or poor health patients the application of cross-over and axillofemoral bypasses are also advisable. PMID- 11484279 TI - [The role of low molecular weight heparin (fragmin) in the prophylaxis and treatment of venous thromboembolism in major abdominal operations]. AB - The goal of the authors was to prepare a thorough survey of the publications about the application of low molecular weight heparines in deep venous thromboses and pulmonary embolism after abdominal surgery. Several low molecular weight heparines have been introduced in clinical practice since 1980. Till then only standard heparin has been used for deep thromboses prevention. Both drugs have the same or similar anticoagulant or antithrombotic effect. Low molecular weight heparines are more widely used than standard heparin because of the possibility to be administered once daily subcutaneously. They don't require a strict laboratory control and can be used in outpatients or home conditions. PMID- 11484280 TI - [Recurrent nodular goiter. Characteristic features and surgical management]. AB - Purpose of the study was to investigate the frequency of the thyroid carcinoma in the recurrent nodular goiter, to analyse the applied operative methods, the observed complications and the obtained early and distant postoperative results. For the period 1985-1996 in the Clinic of Endocrine Surgery were performed 588 reoperations by 577 patients with nodular recurrent goiter, distributed as following: 539 females, aged 14-81 and 49 males, aged 11-69. In 431 cases (73%) was applied medial approach and in 157 cases (27%)--lateral operative approach by the mobilizing of the thyroid recurrence. In the early postoperative period were established the following complications: clinical hypoparathyroidism in 10 patients (1.7%) with calcium level from 1.44-2.01 mmol/l and recurrent nerve paralysis in 28 patients (4.76%), including: in 13 cases unilateral paralysis on the right side, in 9 cases unilateral--on the left side and in 6 cases--bilateral paralysis. The established in 28 patients (4.76%) thyroid carcinoma and the appeared in 59 cases (10%) new recurrence of the basic thyroid disease supported our tactic for radical operative treatment in all patients with nodular recurrent goiter. PMID- 11484281 TI - [Bacterial translocation from the gastrointestinal tract: catalyst of multiple organ dysfunction syndrome]. AB - The abnormal colonization of gastrointestinal tract (GIT), the loss of the intestinal barrier function, the bacterial translocation (BT) are signs of intestinal insufficiency which are supposed to be involved in the pathogenesis of MODS. This worsens the condition or leads to lethal outcome in patients after major abdominal surgery in ICU. The goal of this investigation was to consider the scientific and clinical evidence for the BT role in the pathogenesis of MODS and to present evidence about the advantages and the efficiency of antibiotic combination Amikacin plus Clindamycin as a new therapeutic strategy for the improvement of the outcome in patients with MODS and sepsis. To that purpose patients with diffuse peritonitis of different origin were analyzed. After surgery some patients were left with laparostomy. This gave the possibility for revisions and lavages of the abdominal cavity and for taking material for microbiological analyses. The patients were grouped into two subgroups according to antibiotic treatment: 1st group--combination of usually used antibiotics; 2nd group--Amikacin plus Clindamycin. The second group patients showed good tolerance to this antibiotic combination and good therapeutic effect. PMID- 11484282 TI - [A modification of Freyer's typical prostatectomy]. AB - Adenomectomy in its present-day modality has a millennium-long history. Chronologically, its development, justly or not, is marked by a great number of names among which it is worth noting Sir Peter Freyer (London) and his paper published in 1901, dealing with the radical treatment of the hypertrophied prostate by urinary bladder exposure. E. Fuller (USA) claims that the first practical implementation of the method six years previously is credited to him. Since then, there are no different opinions on the need of total gland enucleation, with some differences remaining in terms of operative access and hemostasis. Over the period 1998-1999, thirty-two adenotomies are performed according to Freyer's original method in the Department of Urology--University Hospital "Alexandrovska". An indwelling catheter with inflated balloon is inserted into the prostate bed for hemostasis without using packs or placing sutures. The obtained results are estimated as good. PMID- 11484283 TI - [Surgical reconstruction options in Poland's syndrome]. AB - We present two cases with Poland's anomaly in a 36 years old man and 27 years old woman and review the possibilities of surgical reconstruction. We performed a one stage chest-wall reconstruction with a prefabricated silicone implant and latissimus dorsi muscle flap in our male patient with chest-wall deformity and partial absence of the pectoral major muscle. Our female patient with absent pectoral major muscle and aplasia of the mammary gland and mammalia underwent a one-stage reconstruction with a prefabricated silicone implant and latissimus dorsi muscle flap and inflatable silicone prothesis for the mound of the breast. PMID- 11484284 TI - [Dermoid, epidermoid and teratoma cysts of the tongue and oral cavity floor]. AB - Most dermoid cysts in the oral cavity floor are derived from epithelial debris or rests, enclaved during midline closure of the bilateral first and second branchial arches. They may also result from ectodermal differentiation of multipotential cells, most likely pinched off at the time of anterior neuropore closure. Congenital epidermoid cysts and teratomata of the tongue may be explained in a similar manner. Only 6.9 per cent of dermoid cysts observed in humans involve the head and neck regions. Treatment consists in complete surgical removal, with excellent prognosis in cases free of complications. This is a report on a rare case of epidermoid cyst, described because of its unusual location and cell population variation of the epithelium lining the cystic cavity. PMID- 11484285 TI - [Laser use in various fields of surgery]. AB - A total of 236 patients presenting various pathological conditions are operated in the Department of General and Operative Surgery of the Medical University- Sofia over the period 1993 through 1998, using laser radiation--CO2-laser in 216 cases, ans Nd-YAG laser--in 20 cases. Both focused and unfocussed laser beam is applied. The superiorities of laser surgery in definite diseases and the results thus far obtained are analyzed. PMID- 11484286 TI - [Esophagojejunostomy after Roux following gastrectomy for cancer of the stomach]. AB - Not big number of cases (17) operated for cardia cancer are subject of retrospective analyzing. Ezophagoejunoanastomosis by termino-lateral plastic, type of Roux is realized in more of cases (9). Termino-terminal junction and this after proximal gastric resection performed on hand, are subject of discussion. PMID- 11484287 TI - [Surgical treatment outlook for gastric carcinoma (a 10-year retrospective study)]. AB - After analyzing clinical material of 159 patients with gastric cancer into Surgical department of the Regional hospital in Dobrich, the authors signified the tendency of later coming for surgery of predominant patients. The part of 66.8% patients coming in 3rd and 4th stage of disease is disturbing. The subtotal resection and gastrectomy like a method of choice are 40.4% with a something more true survival chances. PMID- 11484288 TI - [Surgery as an element of the complex therapeutic approach to myasthenia gravis and thymoma]. PMID- 11484289 TI - [Diagnosis and surgical treatment of adrenocortical carcinoma. A review of the literature and report of two cases]. AB - Adrenocortical carcinomas (ACC) are rare and represent only 0.05 to 0.2% of all cancers. They can be hormonally active, appearing clinically by one or more endocrine syndromes, or can be hormonally non-active. Commonly most of the cases with AC are diagnosed when the neoplastic process have spread out of the suprarenal gland (stage III or IV). Computed tomography and magnetic resonance imaging are the most useful diagnostic methods of ACC but the latter is more accurate, especially in estimation of the local invasion of the tumor. Surgery is the main and the most effective method for treatment of both primary and recurrent ACC, and in selective cases--of metastasis. The chemotherapy with mitotane has a limited role and is indicated in cases of inoperable ACC (primary or recurrent) and/or presence of metastasis. Two cases of ACC treated in our department are reported. Case I: a 26-years-old female with ACC in stage II, which was diagnosed incidentally by ultrasound investigation for other consideration. Although the urine levels of free cortisol were elevated, the woman had no endocrine symptoms. Case II: a 50-years-old female with a second recurrence ACC appeared 5 years after a resection of the first recurrence tumor and 6 years after an operation for the primary tumor. Problems in diagnose and surgical treatment of these cases are discussed. PMID- 11484290 TI - [Carcinoma of the stomach - our experience with surgical treatment]. AB - Carcinoma of the stomach is among the commonest malignancies of the gastrointestinal tract regardless of the permanent tendency of its diffusion to decrease, observed in the last 25-30 years. The readily accessible methods of diagnosing the disease contribute greatly to its early detection. However, owing to diverse causes, in over 70 per cent of cases the diagnosis is usually made as late as in the advanced III-IV stages. The latter circumstance preordains largely the unfavourable long-term results of the treatment undertaken where surgery plays a major role. It is the purpose of this study to analyze the surgical strategy and tactics currently used in the management of gastric carcinoma. Over a 5-year period (Jan 1995 through Dec 1999), in the Chair of General and Operative Surgery of the Medical University--Sofia a total of 184 gastric carcinoma patients, including 116 men (63.1%) and 68 women (36.9%) with age ranging from 23 to 80 years, undergo operation. Diagnosing is based on past history, physical, laboratory and x-ray data, but first and foremost on evidence from FGS and histological assessment of biopsy material (carried out in all patients). With a view to precise preoperative staging of the lesion, roentgenoscopy + roentgenography of lungs, USD and CAT of the abdominal organs are also done. The following intervention are performed: gastrectomy 18 (9.8%), upper pole resection 43 (23.4%), subtotal resection of stomach 4 (2.2%), prosthetic replacement of cardia 8 (4.3%), derivations 22 (11.9%), and explorative laparotomies. Combined subtotal gastric resections of gastrectomies are necessitated in 73 patients (39.7%) because of carcinomatous infiltration of contiguous organs and/or presence of liver metastases. Morbidity involves 29 patients (15.7%) with lethality amounting to 16 (8.7%). The long-term postoperative results are discussed under a separate heading. Operative treatment of gastric carcinoma patients is the only chance of survival. The scope of indication for more aggressive surgical interventions, including combined resections and gastrectomies, are broadened leading in turn to a considerable reduction of the proportion of explorative laparotomies. PMID- 11484291 TI - [Colorectal carcinoma and molecular genetics]. AB - In the last few years, problems relating to genetic determination of colorectal carcinoma are comprehensively discussed by a number of authors. The new elaborations concerning the familial polypoid and carcinoid syndromes discovered, as well as molecular researches along this line answer a great number of questions posed in connection with substantiating the old idea about many-staged cancerigenesis. As much as 94 per cent of colorectal carcinoma cases are assigned to the sporadic cancer diagnosis category. Familial polyposis accounts for 1 per cent of all neoplasms of the colon, while the remainder 5 per cent make part of diverse familial carcinoid syndromes. Most of the data on genetic etiology of colorectal carcinoma so far elucidated are presented in a summed up fashion. PMID- 11484292 TI - [Successful treatment of consecutive chest surgery complications]. PMID- 11484293 TI - [Case of an HIV-patient with advanced form of perianal hidradenitis]. PMID- 11484294 TI - Tips for passing the all-important exam. PMID- 11484295 TI - General dental practice in the hospital. Introduction. PMID- 11484296 TI - A focus on the institutionalized aged and special care patient for today's practice. AB - Dentists must understand the growing institutionalized-aged and special-needs population, the places wherein they reside, and the unique challenges of access that confront both the patient and dentist. This article discusses governmental regulation and legislation of long-term-care facilities and outlines professional duties and requirements of dentists who care for residents of such facilities. It will also cover the treatment needs of this population and the venues available to the hospital-trained dentist. PMID- 11484297 TI - Developmental disabilities and understanding the needs of patients with mental retardation and Down syndrome. AB - This article provides the dentist with a background on developmental disabilities, services, and issues related to statutory and regulatory requirements, with a focus on mental retardation and Down syndrome. Down syndrome has mental retardation as a component, yet requires additional emphasis because of its prevalence and associated craniofacial manifestations. Also discussed are treatment considerations. PMID- 11484298 TI - Developmental disabilities: epilepsy, cerebral palsy, and autism. AB - This article provides the dentist with a review of the three developmental disabilities that do not have mental retardation as a diagnostic component: epilepsy, cerebral palsy, and autism. Discussion focuses on diagnostic criteria and other dental and medical considerations. A greater understanding of developmental disabilities allows the dentist to offer care in the dental office when feasible or to understand and develop referral relationships with colleagues who utilize the hospital operating room to provide comprehensive care. PMID- 11484299 TI - Integrating hospital dentistry into the general dental practice. AB - Societal trends and medical advances have resulted in increased numbers of physically and/or psychologically challenged individuals living within our communities. For some of these individuals, hospital dentistry with general anesthesia provides the only means by which general dental services may be provided. This paper presents considerations of significance to the general dentist wishing to incorporate outpatient hospital dentistry into his or her private practice. PMID- 11484300 TI - Antibiotic supplementation of bone allografts: a review. PMID- 11484301 TI - Clinical and economic impact of low-molecular weight heparins in managed care. Introduction. PMID- 11484302 TI - Overview of enoxaparin in the treatment of deep vein thrombosis. AB - Standard medical treatment for patients with acute venous thrombosis is antithrombotic therapy. Following a historical overview of the evolution of heparin therapy to include low-molecular weight heparin (LMWH), the pharmacokinetic properties of enoxaparin are described. Therapeutic advantages of LMWH over unfractionated heparin (UFH) are also discussed, including once- or twice-daily subcutaneous dosing, reduced hospital stays, elimination of therapeutic monitoring for most patients, and possibly less bone density loss. Studies have demonstrated that enoxaparin is at least equivalent to UFH with regard to efficacy and safety. Opportunities for future study include evaluation of enoxaparin's efficacy for the prevention of deep vein thrombosis within high risk groups and for the treatment of thrombosis in such conditions as pregnancy, cancer, obesity, and renal insufficiency, and in children. PMID- 11484303 TI - Outpatient-based treatment protocols in the management of venous thromboembolic disease. AB - Clinical trial data are demonstrating that the safety and efficacy of treating deep vein thrombosis (DVT) on an outpatient basis with low-molecular weight heparin (LMWH) are equal to inpatient care with unfractionated heparin (UFH). Moreover, LMWH therapy offers multiple advantages, including improved bioavailability at lower doses, reduced heparin resistance, a longer half-life, and potentially, less bleeding. Although these potential advantages are acknowledged, patient selection and risk stratification criteria for patients being considered for LMWH therapy are discussed less frequently; nonetheless, they deserve careful consideration. The protocol utilized by Lovelace Health Systems, a staff model health maintenance organization, features an outpatient based DVT treatment program that includes exclusionary risk factors for outpatient treatment of venous thromboembolic disease (VTE). Evidence from clinical findings at Lovelace suggests that patients without absolute exclusionary risk factors can successfully be treated with LMWH as outpatients. Specific study results show that 61% of patients were eligible for outpatient therapy and had fewer recurrences of VTE (1.9%) than those comorbid matched patients who were hospitalized with traditional UFH therapy the previous year (4.1%). These findings suggest that LMWH is both safe and efficacious in the home treatment of VTE in a managed care setting when established patient selection criteria and risk stratification strategies are carefully maintained. PMID- 11484304 TI - Deep vein thrombosis and pulmonary embolism in the perioperative patient. AB - Approximately 500,000 cases of deep vein thrombosis (DVT) and pulmonary embolism (PE) occur in the United States each year. Of those patients who suffer a massive PE, 70% die within the first hour of symptom onset. Thus, early and aggressive intervention is essential. Clinical evaluation of patients is key in assessing clot risk and is aided by a variety of screening devices, with venography as the gold standard. Patients who undergo hip and knee arthroplasty are at highest risk for DVT and PE. However, appropriate prophylaxis can reduce the incidence significantly. Although standard low-dose heparin is considered to be ineffective, positive experience with the administration of the low-molecular weight heparin (LMWH) enoxaparin, because of the speed of its efficacy in postoperative patients at high risk for DVT, has been reported. The dosage of enoxaparin is weight-adjusted and is sometimes combined with warfarin. Tools for risk-factor assessment and suggested prophylactic regimens for patients undergoing total hip and knee replacement are presented. PMID- 11484305 TI - Economics of low-molecular weight heparins. AB - Low-molecular weight heparins (LMWHs) have become attractive alternatives to standard unfractionated heparin (UFH) for the treatment and prophylaxis of deep vein thrombosis (DVT), and for the management of acute coronary syndrome (ACS). The economic impact of the use of LMWHs has been studied in randomized controlled studies, in demonstration projects in managed care institutions, and in decision models. These studies provide valuable insight into the ways LMWHs can be economically attractive despite higher per unit costs compared with UFH. For the treatment of DVT, the cost benefit of using LMWHs results primarily from the cost shifting from inpatient to outpatient care, and might be limited by the eligibility of patients for outpatient management. In contrast, the attractiveness of LMWHs for ACS and DVT prophylaxis hinges on the increased effectiveness of LMWHs compared with standard UFH. PMID- 11484306 TI - Address to the House of Delegates May 5, 2001. PMID- 11484307 TI - . . . and things that go bump in the night. PMID- 11484308 TI - Skin cancer. PMID- 11484309 TI - Common sun-induced cutaneous lesions of the head and neck: what dentists should know. PMID- 11484310 TI - Malignant melanoma. PMID- 11484311 TI - A review and update of intraoral lichen planus. PMID- 11484312 TI - Oral and maxillofacial pathology case of the month. Pleomorphic adenoma (PA). PMID- 11484313 TI - Origins of Argentinean dentistry and development of teaching. AB - The history of the legal practice of dentistry in the Rio de la Plata region is given. The first diplomas were given in 1891 by the Medical Tribunal, before the creation of the dental school in Buenos Aires. The inauguration of teaching courses and the different teaching plans, as well as the biography of the teachers who initiated the teaching in Argentina are presented as are the creation of the School of Dentistry and the appearance of the three schools of the College of Medical Sciences in the monumental new building. Portraits of the teachers Etcheparborda, Pereira, Erausquin and Guardo, crafters of the old school are presented, together with those of the administrators of the university complex and the College, Profs. Dr. Jose Arce and Ricardo C. Guardo. PMID- 11484314 TI - Edgar Buchanan: dentist and popular character actor in movies and television. AB - Edgar Buchanan, D.D.S., pursued a diverse mix of careers during his lifetime: as he practiced dentistry, he also worked as a popular film and television actor. Although he eventually relinquished a full-time dental practice for acting, he continued his commitment to clinical dentistry. Acting in 100 films and four television series across a 35-year span (1939-1975). He personified a scheming, yet well-meaning rustic who specialized in "cracker-barrel" philosophy. Typically, he was cast in classic western movies as a bewhiskered character actor. In several films he played a frontier dentist who was always portrayed in a sympathetic and authentic manner. His unique gravelly voice, subtle facial expressions, folksy mannerisms and portly build enabled Buchanan to step into a wide variety of character roles. His most memorable television role was in the classic situation comedy, "Petticoat Junction," (1963-1970), where he played Uncle Joe, a folksy, lovable, free-loader whose many entertaining schemes created chaos. PMID- 11484315 TI - Gleanings about dentistry from the world of literature (twenty-four in a series). PMID- 11484316 TI - Connections--dentistry and medicine. PMID- 11484317 TI - How a dentist's name became a synonym for a life-saving device: the story of Dr. Charles Stent. AB - Stents have been used in numerous medical disciplines, as well as in oral surgical procedures. Uses range from rebuilding mandibles and constructing new ureters, to keeping coronary arteries patent after angioplasty. The earliest use of the word "stent" to describe this item was in 1916, when a Dutch plastic surgeon described how he used a dental impression compound as a matrix around which to form tissue in the process of rebuilding a shattered face. What is generally unknown is that the word "stent" derives from the name of an English dentist, who invented this impression compound in 1856. PMID- 11484318 TI - Charles G. Pease, DDS, MD: anti-tobacco crusader and clean life advocate. AB - Dr. Charles Giffin Pease, a colorful, feisty, New York City reformer practiced both dentistry and medicine and lectured in various dental and medical schools. At several New York dental institutions, he taught oral surgery, materia medica, therapeutics and pathology. From the age of 12 until his death (a 75 year span) Pease waged an unrelenting war against "everything that harms the human race." He wrote and lectured unceasingly railing against what he believed to be life damaging behaviors. These included: tobacco use; alcohol, coffee, tea and ginger ale consumption; corset wearing; vinegar, meat, cocoa, chocolate and condiment intake, and even the licking of artificially flavored lollipops. While he was adamantly opposed to all of these practices, he invested his strongest efforts campaigning against the manufacture, sale and use of tobacco products; and he emphatically supported clean living causes. His anti-tobacco stance was based chiefly on moral, ethical and religious precepts. Additionally, he expected community role models (public health officials, clergymen, physicians and dentists) to become involved in this vital health issue. When publicly presenting his cause, he used a moralistic, judgmental approach. Almost single handedly, Pease spear-headed legislation that prohibited smoking in subways, elevated trains, hotels and restaurants. PMID- 11484319 TI - Dentistry on stamps. PMID- 11484320 TI - The "Amex" cast aluminum denture of World War I. AB - In 1917-18, the U.S. Army revived a denture technique first introduced in 1866 by Dr. James Baxter Bean, the Confederate dental surgeon who established the first military maxillofacial hospital trauma ward in Atlanta, Georgia, during the American Civil War--the cast aluminum wartime denture. PMID- 11484321 TI - Bush's no-win choice. PMID- 11484322 TI - The great cell debate. PMID- 11484323 TI - One giant step for mankind. PMID- 11484324 TI - A quick dip in a dirty pool. PMID- 11484325 TI - Mounting the slippery slope. PMID- 11484326 TI - Trichinellosis in the Slovak Republic. AB - The occurrence of trichinellosis in farm or wildlife animals in some areas pose the permanent risk of infection in humans. In Slovakia, where only the sylvatic trichinellosis exists, the humans trichinellosis is sporadic. The most common cause of infection is wild boar meat. Unusual local eating customs may also favour an outbreak of disease in humans. The outbreak of trichinellosis in Slovakia in 1998, when 336 people were affected, was of the same kind. Trichinella britovi was the agent of the infection. PMID- 11484327 TI - Diagnosis and epidemiology of Trichinella infections in wildlife in The Netherlands. AB - Trichinella infections in foxes and wild boars were studied to determine the prevalence of infection in wildlife in the Netherlands. Muscles of 429 forelegs of foxes (Vulpes vulpes) and diaphragms of 11 wild boars (Sus scrofa) were artificially digested. Single larvae of Trichinella were identified at species level using random amplified polymorphic DNA (RAPD-PCR). In addition, an ELISA using ES antigen was used to test serum samples of 458 wild boars. The prevalence of Trichinella in foxes ranged from 3.9% in the the eastern part of the country, 13.1% in the central part of the country and 1.3% in the most western part of the country. Trichinella larvae of foxes were identified as T. britovi. In most samples, identification of larvae did not show reproducible results. The serological prevalence of Trichinella infections in wild boars was 6.8%. Wild boar populations are located in the central and in the southern part of the country. Trichinella larvae of wild boar were identified as T. spiralis. These results show that two Trichinella species are involved in the epidemiology of trichinellosis among wildlife. On the basis of previous reports, the present results suggest that the prevalence of Trichinella infection in wildlife is increasing in the last 20 years. PMID- 11484328 TI - Epidemiology of human and animal trichinellosis in Italy since its discovery in 1887. AB - The epidemiology of trichinellosis in Italy is characterised by a sylvatic cycle present only on the mainland. The domestic cycle probably never existed, though a domestic focus occurred on the island of Sicily between 1933 and 1946. The red fox is the main reservoir, with the prevalence of infection ranging from 0.0% in lowlands to 60% in the Alps. The main etiological agent is Trichinella britovi. Trichinella pseudospiralis has been detected in two birds. From 1948 to March 2000, trichinellosis was diagnosed in 1,347 persons, who acquired the infection in 21 outbreaks. PMID- 11484329 TI - Trichinellosis: now and forevermore? AB - The re-emergence of the zoonotic importance of trichinellosis reminds us that this parasite's unique biological features serve it well in helping it to persist in the face of 150 years of efforts to eliminate it from the food chain. Because of Trichinella's re-emergence as an important public health risk, and the consequent stimulation of research, our knowledge of the parasite has grown, especially on the complexity of its biology and epidemiology. This overview will highlight the major features of this parasite's adaptive traits and the human factors that account for its re-emergence in many regions, along with the actions of the International Trichinellosis Commission to address these issues. The attractiveness of Trichinella as a research subject for study of the parasitic lifestyle will also be emphasized, including areas for future research. PMID- 11484330 TI - Mucosal responses to infection with Trichinella spiralis in mice. AB - Infections with T. spiralis in mice elicit strong inflammatory responses. The nature and control of these responses, and their relationship to the process of worm expulsion, have been debated for many years. Many components of inflammation are, like worm expulsion, T cell-dependent, but some are not. The paper describes novel observations on Paneth cell responses to infection in immunologically normal mice and in a variety of T cell-deficient mice. Responses occurred normally in nu/nu and scid/scid mice but not in beta/delta knock out mice incapable of generating cells with functional TCR. However all of these mice failed to expel worms in the pattern seen in immunologically normal controls. These data are incorporated into a discussion of the causal relevance of intestinal inflammatory changes to the process of worm expulsion. PMID- 11484332 TI - A comparison of antigenic peptides in muscle larvae of several Trichinella species by two-dimensional western-blot analysis with monoclonal antibodies. AB - The antigens recognised by mAb US5 specific to 53 kDa glycoprotein (gp 53) in T. spiralis L-1 muscle larvae (TSL1) antigens, mAb US9 specific to gp 53 in TSL1 from all encapsulated species and mAb US4 specific to a tyvelose containing tetrasaccharide present in TSL1, were investigated in crude extracts from muscle larvae of T. spiralis, T. nativa and T. britovi by 2D-electrophoresis and western blot. At least four proteins of different p1 were recognised by mAb US5 on T. spiralis antigens. Recognition profile of mAb US9 on T. spiralis antigens exhibited some variation with regard to that of the US5. Polymorphism was apparent in gp 53. High reactivity was shown by the mAb US4 with the three species. PMID- 11484331 TI - Interleukin mRNA changes in mast cells stimulated by TSL-1 antigens. AB - In this work we analyzed by RT-PCR, the mRNA changes for IL-4, IL-10, TNF and IFN (induced by TSL-1 antigens in a rat mast cell line (HRMC) with mucosal characteristics. The data obtained showed an increase of 65 and 52% in mRNA expression for IL-4 and TNF respectively and a decrease of 59 and 55% in mRNAs for IFN gamma and IL-10. Our results suggest that TSL-1 antigens induce the release from MC of regulatory molecules, such as IL-4 by an IgE independent mechanism. Our data also provides important information related to the ability of MC to participate not only in the effector phase against the infectious agents, but also in the orchestration of the immune response by the host against parasites. PMID- 11484333 TI - Possible presence of common tyvelose-containing glycans in Trichinella L1 larvae and embryonated eggs of several nematodes. AB - A monoclonal antibody (mAb US4) recognising an epitope containing tyvelose within the T. spiralis L-1 muscle larvae (TSL-1) antigens was tested in western-blot against various antigenic preparations from different stages of the following nematodes: T. spiralis (L1, adult), T. muris (egg, L1, L3, adult), Ascaris suum (egg, adult), Toxocara canis (egg, adult), Anisakis simplex (L3) and Haemochus contortus (egg). Positive reaction was present in antigen preparations from L1 larvae of T. spiralis and T. muris and from embryonated eggs of T. muris, A. seum, T. canis and H. contortus. PMID- 11484334 TI - Detection of circulating and fecal Trichinella spiralis antigens during experimental infection using monoclonal antibodies against the new born larvae. AB - Different assays to detect antigens of Trichinella spiralis during current infection have been standardized, although sensitivity values have been the main limitation to use them as routine diagnostic test. We report the production and use of monoclonal antibodies (MAbs) against the new born larvae which recognize both somatic and metabolic antigens from adult and muscular larvae (ML). We used two IgG3 MAbs (4B1, 4B2) and two IgG2a (2D3, 2D4) to detect antigens during experimental infection. All MAbs detect fecal antigens starting the second to third week post infection (wpi), although it was less clear with MAbs 4B1 and 4B2, while circulating antigens were detected from third to fourth wpi. Thus, the recognition of shared antigens among T. spiralis developmental stages can be used for early diagnosis of trichinellosis. PMID- 11484335 TI - Assessment of efficacy and safety of various adjuvant formulations with a total soluble extract of Trichinella spiralis. AB - Trichinellosis, a re-emerging zoonosis in several countries and pig, is the main species responsible for its transmission to human. Vaccination of swine could be an alternative to prevent the risk of human contamination. In order to develop an efficient and safe inactivate vaccine, the choice of the adjuvant is an important issue. The aim of this study was to develop and select potent and safe adjuvants by screening them in an experimental model with a crude soluble antigen from L1 muscular larvae (ML) of Trichinella spiralis (Ts). The efficacy was checked by the quantification of specific antibody levels. Specific and non-specific IgE antibody levels were also assessed. Safety was checked by the assessment of the local reaction at the injection site. Various Montanide ISA adjuvant formulations including water in oil, oil in water and multiphasic emulsions, but also nanoparticles or microbeads were tested. The results clearly showed differences between the antibody responses induced by the adjuvants and demonstrated the necessity to use an adjuvant to obtain a specific IgG (IgG1 or IgG2a) response directed against the total soluble extract of Ts. All the formulations enhanced the humoral immune response. The origin of the oil contained in the emulsions played an important role on the efficacy. Indeed emulsions based on mineral oils were more efficient than those based on metabolisable oils. However it was linked with stronger local reactions. Multiphasic and oil in water emulsions but also nanoparticles failed to induce IgG2a antibody levels. Microbeads and water in oil formulations based on mineral oils were more efficient. This experimentation allowed then the selection of several adjuvants which efficacy will be further investigated by a challenge test and an analysis of the cellular populations involved in the mechanism of the immune response. PMID- 11484336 TI - Comparison of IgM, IgG1 and IgG2 responses to Trichinella spiralis and Trichinella britovi in swine. AB - Pigs infected with T. spiralis and T. britovi were followed by double (IgG) and triple antibody ELISA (IgG1, IgG2 and IgM) during a 12-week-period. Specific IgG and IgG1 responses were similar and showed a significant relation with the infecting doses and intensity of infection. Response to T. britovi was slightly lower than in groups infected with the same dose of T. spiralis. IgG2 response was weak and almost undetectable in the lowest infected pigs, but relationship with the intensity of infection was unclear. IgM antibodies showed rapid but transient increases, generally simultaneous to peaks of IgG response. PMID- 11484337 TI - Detection of coproantigen in early trichinellosis. AB - Trichinellosis has become undoubtedly worldwide in distribution. Its diagnosis relies largely on the serodiagnostic procedures which are of great value but unfortunately miss the enteric phase. This could be a serious diagnostic problem in the absence of corresponding epidemiological data and typical symptoms and signs of the disease. In this study the possibility of coproantigen detection, as an early diagnostic aid in trichinellosis, was investigated in mice experimentally infected with Trichinella spiralis. A modified double sandwich ELISA was developed using polyclonal antibodies raised in rabbits and guinea pigs against larval somatic antigens. The first detection of coproantigen was as early as the first day post infection, gradually increasing to reach its peak on the seventh day and then decreasing to disappear completely on the third week post infection. Another test, the coagglutination test (Co-A) was used, and this test confirmed the previous results. The finding of this study suggest that the coproantigen detection could be exploited to confirm ongoing early Trichinella spiralis infection. This fast and easy to use diagnostic method should improve the early infection in human. PMID- 11484338 TI - Remembrance of past images of Trichinella. AB - The development of achromatic microscopy and the invention of photography were contemporaneous with the earliest investigations on trichinellosis. The former was more important than the latter to 19th century studies on Trichinella. A selection of images, diverse but not comprehensive, is presented to illustrate the early history of trichinellosis. PMID- 11484339 TI - The use of a synthetic antigen for the serological diagnosis of human trichinellosis. AB - Hosts infected with Trichinella produce antibodies specific for an epitope common to the TSL-1 family antigens. This epitope contained uncommon terminal 3, 6 dideoxy-D-arabinohexose (so called tyvelose) residues. The disaccharide moiety was synthesized and an immunodiagnostic assay was developed, which was specific and sensitive in swine trichinellosis. We aimed to verify the specificity and sensitivity of this immunodiagnostic test in human trichinellosis. 15 sera from normal subjects, 12 from patients with other parasitic diseases and 50 from trichinellosis patients were tested. Indirect enzyme linked immunosorbent assay (ELISA) for specific IgG and an amplified ELISA for specific IgE were performed using beta-tyvelose-GalNAc-bovine serum albumin (BSA) disaccharide conjugate or T. spiralis muscle larvae excretory/secretory (E/S) products, as antigens. Neither control sera nor other parasitic infection sera resulted positive both for IgG and IgE when synthetic or E/S antigens were used. In trichinellosis patient sera, specific IgG were present in 100% of cases, irrespective of the antigen used, but whereas specific IgE were detected in 78% using E/S antigens, a 100% positivity rate was obtained, using the beta-tyvelose-BSA conjugate. PMID- 11484340 TI - Diagnosis of human trichinellosis: pitfalls in the use of a unique immunoserological technique. AB - Serum samples belonging to three outbreaks in Argentina (47 patients) taken at different times post-ingestion were analysed employing IIF and ELISA simultaneously. Results show that: a) the number of patients diagnosed by a unique technique, especially by ELISA (31 patients), was lower than the one obtained by the simultaneous use of both assays (38 patients); b) four patients out of the seven diagnosed by a unique technique were negative by the other assay over the period of time evaluated. Therefore, it can be concluded that the use of a sole immunoserological technique can not only lead to the delay in the detection but also to the misdiagnosis of this parasitic infection. PMID- 11484341 TI - Clinical appraisal of patients and detection of serum antibodies by ELISA and CIA tests in late periods of Trichinella sp. invasion. AB - Results of our studies using ELISA and competitive inhibition assay (CIA) tests fully confirmed the previously experienced trichinellosis and reflected persistent stimulation of antibody production due to the antigen release from Trichinella larvae, which had survived longer and undergone gradual destruction in the muscles. The studies proved that the tests complement each other, yielding concordant results in 86.7% of cases. Due to its higher specificity, the CIA test can help in interpreting pathological signs/symptoms and in evaluating humoral response activity at late and distant in time periods following the invasion. PMID- 11484342 TI - Class specific antibody responses to newborn larva antigens during Trichinella spiralis human infection. AB - A follow-up study of the class antibody responses to newborn larva (NBL) antigens in individuals involved in an outbreak of human trichinellosis was carried out by ELISA assays. The data showed that similar kinetics of antibody responses of different magnitude developed in trichinellosis patients; it was low by week 3, a peak raised by week 5 and decreased from week 7 up to the end of the study. The IgA-ELISA assay was the most sensitive and specific while the IgM was the least sensitive and specific. IgA antibodies to NBL antigens were detected in 80% of patients while IgE, IgG and IgM responses were observed in 44, 31 and 19% of the patients by week 3, respectively. From weeks 5 to 7, IgA antibodies were found in 89 to 100% of the patients while lower percentages (0-82%) were found for the other isotypes. Reactivity of IgA, IgE, IgG and IgM to NBL antigens decreased from week 37 to 57 after infection (0-38%). These results suggest that detection of IgA antibodies may be useful for early diagnosis and epidemiological studies in human trichinellosis. PMID- 11484343 TI - The stage-specificity of the IgA response to newborn larva and TSL-1 antigens of Trichinella spiralis in humans infected with the parasite. AB - Characterization of human IgA responses to newborn larva (NBL) and TSL-1 antigens was carried out by ELISA assays. Relevant and differential IgA antibody responses to these antigens were detected in humans infected with T. spiralis. The inhibition ELISA results showed that the IgA response to NBL antigens was inhibited significantly by both NBL and TSL-1 antigens and to a lesser extent when phosphorylcholine (PC) was used as inhibitor. In contrast, the IgA response to TSL-1 antigens was inhibited by the homologous antigen and to a lesser extent by the NBL and PC. Thus, the early IgA antibodies developed in trichinellosis patients contained a portion of IgA antibodies directed to PC which is present in TSL-1, A and NBL component. Another portion of antibodies to NBL are directed to other common non-defined epitopes present in TSL-1 and NBL antigens. All together these results suggest that the IgA response to common epitopes in antigens of both stages of the parasite may be useful for early diagnosis and epidemiological studies of human trichinellosis. PMID- 11484344 TI - History of trichinellosis surveillance. AB - The origin of trichinellosis, which existed in ancient times as testified by the discovery of parasite larvae on an Egyptian mummy, unfolded in several stages: discovery of encapsulated larvae (in the 1820s), identification and scientific description of these larvae (Paget & Owen, 1835), followed by experimental infestations of animals (dogs, pigs, rabbits, mice) or of humans as from 1850. The main occurrences of trichinellosis were followed with particular attention in Europe (Germany, Denmark, France, etc.) and in the United States of America at the end of the XIXth century. They affected numerous domestic animal species (pigs, horses, etc.) or wildlife and humans. Germany paid the heaviest toll with regard to the disease in humans, between 1860 and 1880, with several thousands of patients and more than 500 deaths. Different trichinellosis surveillance systems were set up in the relevant countries in the 1860s. In humans, this surveillance was carried out on affected living patients by a biopsy of the biceps muscles and subsequently by an analysis of eosinophilia (1895). In animals, surveillance was for a long time solely based on postmortem examination of the muscles of the affected animals. This method was used for the first time in 1863 in Germany, and from the 1890s, on several hundreds of thousands of pigs in Europe or in the United States of America. PMID- 11484345 TI - A follow-up study of the human class and subclass antibody response developed against the adult stage of Trichinella spiralis. AB - We report the analysis by ELISA of class and subclass antibody response against a total soluble extract from T. spiralis adult stage (TSE-A) during a year after the infection in 17 symptomatic trichinellosis patients (SI) and five asymptomatic individual (AI) involved in an outbreak of trichinellosis occurred in the State of Mexico. Serum samples from 20 healthy individuals (HI) and 24 patients with other parasitosis were included as control. All SI showed a polyisotypic antibody response against the TSE-A, during the infection. Higher response of IgA, IgE, IgM were detected in SI during the acute phase of the infection, but only IgE remained at high levels all along the infection. None or a lower reactivity against TSE-A was observed in sera from AI and from HI. Some patients with trichuriosis and ascariosis showed a higher cross-reactivity, against TSE-A when IgG and their subclasses were analyzed. PMID- 11484346 TI - Antigen recognition by IgG4 antibodies in human trichinellosis. AB - The antibody isotype response to Trichinella spiralis excretory/secretory (ES) products of muscle larva was examined using sera from patients with confirmed trichinellosis. Using Western blots we identify components of the ES antigen that are recognized by IgM and IgG antibodies. A 45 kDa component was strongly recognized by different antibody classes and subclasses. We observed a 45 kDa specific IgG4 response that was detected exclusively using sera of patients with trichinellosis and not of patients with echinococcosis, filariasis, cysticercosis, ascariasis, strongyloidiasis or toxocariasis. These results are relevant for the diagnosis of human trichinellosis. PMID- 11484347 TI - New aspects of clinical pathology and electro-physiological muscle disturbances in patients with history of trichinellosis. AB - Clinical studies performed in 44 patients, one, three, four, six or seven years after they had experienced trichinellosis demonstrated the persistence of various general and motor ailments in 88.7% of the patients. The persisting for many years antibodies against E/S antigen of Trichinella in 86.4% of examined patients seem to reflect chronic stimulation by the larvae which, as shown by parasitological and histopathological tests, survived longer and underwent gradual destruction in muscles. Bio-electric disturbances in muscles were most frequently of a mixed type, with prevalence of a neural type record. These observations illustrate a disturbed function of motor neurones and of impulse transmission at the myoneural junction in patients chronically infected with Trichinella. PMID- 11484348 TI - Trichinella murrelli: pathological features in human muscles at different delays after infection. AB - The authors describe the pathological aspects of muscles of three patients infected with Trichinella murrelli. Biopsies were carried out at various intervals. Six weeks after infection, the muscular larvae were not encapsulated whereas encapsulation was seen 10 weeks after infection. Six years after infection, the larvae were still alive in a nurse cell surrounded by a very thick capsule. Fourteen years after infection, cuticular larvae remnants were seen in degenerating nurse cells. The late encapsulation of Trichinella murrelli in human muscles could explain some clinical differences noticed during the outbreak during which these three patients were infected. PMID- 11484349 TI - Congenital trichinellosis? Case report. AB - A large trichinellosis outbreak in the Slovak Republic caused by the species Trichinella britovi and affecting 336 people also affected a pregnant woman. The mother was infected in the 10th week of pregnancy and was treated with mebendazole. On her own request abortion was performed in the 22nd week of pregnancy. Medium IgM and high IgG anti-Trichinella antibody titres were found. The placenta, body cavities liquid, tissues and organs of the foetus contained 0.02-30 larvae per gram of tissue, measuring 0.68 +/- 0.05-1.17 +/- 0.07 mm, with blurred inner structure. Immunocytochemical examination identified Trichinella larvae that infected the foetus in the early stage of development. PMID- 11484350 TI - Aspects of clinical features, diagnosis, notification and tracing back referring to Trichinella outbreaks in north Rhine-Westphalia, Germany, 1998. AB - 52 cases of human trichinellosis were notified from 11 towns in North Rhine Westphalia from November 1998 to March 1999. After non-typical symptoms in the enteral phase, fever, muscular ache, headache, oedema, disorder of vision and rash occurred in the parenteral phase. Trichinellosis was serologically confirmed by ELISA, IFAT or western blot. Raw sausage and minced meat produced from raw pork could be determined as probable source of infection with 44 and eight notified cases, respectively. Whereas questionable raw sausage was not available for examination, frozen minced meat from the second outbreak could be secured in households of infected people. Larvae were isolated from minced meat and were identified by PCR as Trichinella spiralis. Tracing back to the source of infection was difficult because of the long time between clinical symptoms, laboratory diagnosis and notification as well as complex trade routes for pork and its products. Trichinella cases emphasize the necessity to meet the prescribed slaughter inspection and to guarantee a reliable prove of origin for meat products especially in view of specific consumer habits, i.e. the consumption of raw meat. PMID- 11484351 TI - An expected outbreak of human trichinellosis for the consumption of horsemeat. AB - In late February 1998, an outbreak of human trichinellosis occurred in the town of Piacenza (northern Italy) among 92 persons who had eaten raw horsemeat. The source of infection was a horse imported to the abattoir of Brescia one month previously. Although the horse had been found to be positive for trichinellosis upon routine examination, the head of an uninfected horse was exchanged with the head of the infected animal, which was mistakenly placed on the market. PMID- 11484352 TI - Improving bioavailability and anthelmintic activity of albendazole by preparing albendazole-cyclodextrin complexes. AB - The bioavailability and anthelmintic activity of albendazole-cyclodextrin complexes (ABZ-CDC) compared to albendazole suspensions in carboxymethylcellulose (ABZ-CMC) was assessed in a mouse model for Trichinella infections. Swiss CD-1 mice experimentally infected with T. spiralis were treated with both formulations against enteral (adult worms) and parenteral (migrating and encysted larvae). Oral bioavailability was assessed in age matched mice treated with 50 mg/kg of both formulations. The anthelmintic effects and plasma concentration of the active metabolite albendazole-sulphoxide (ABZSO) enantiomer (-) were significantly increased following administration of ABZ-CDC in relation to ABZ CMC. PMID- 11484353 TI - The efficacy of flubendazole against Trichinella spiralis in swine. AB - A trial was undertaken to assess the efficacy of flubendazole against experimentally induced trichinellosis in pigs when given mixed with food at different dose rates. At the start of the experiment, 48 pigs were infected with approximately 35,000 larvae of Trichinella spiralis, afterwards the animals were distributed in six groups. The first three groups were used to test the activity of the drug against adult stages. Other three groups served to define the efficacy against muscle larvae. The dose rate of flubendazole varied according to the purpose of the experiment (from 8 mg/kg up to 62 mg/kg). At the end of the trial, the pigs were necropsied and the number of adults and larvae in predilection muscles were determined. At a dose rate of 8 mg/kg flubendazole supplied with food during eight days was 100% effective against adults. At a dose rate of 31 mg/kg flubendazole administered for 14 days was 72.35% effective. Treatment with higher dosages of flubendazole (62 mg/kg) resulted in increased efficacy (87.77%) against the parasites. The biological assay performed with larvae from the muscle samples originating from the pigs treated with 62 mg/kg showed that at least half of the larvae were not infective. PMID- 11484354 TI - Efficacy of flubendazole and albendazole against Trichinella spiralis in mice. AB - Efficacy of flubendazole and albendazole against Trichinella spiralis in mice were studied. ICR mice were experimentally infected with Trichinella spiralis and treated with either flubendazole (FBZ) or albendazole (ABZ) at four different stages of the parasite life-cycle. Oral administration of either FBZ or ABZ at 20 mg/kg and 50 mg/kg on 2 h, 8 h and 24 h (pre-adult stage) after infection eliminated 94.7-100% of adults as determined at necropsy on day 7 post infection (p.i.) and 96.9-100% of larvae on day 45 p.i. FBZ was more effective than ABZ against adult T. spiralis (at 2 to 6 days p.i.), when treated with a dosage of 20 mg/kg for 5 consecutive days (99.4% and 46.0% reduction with respect to the control group). Against migrating larval T. spiralis, FBZ was more effective than ABZ at 20 mg/kg for five consecutive days (on days 11-15 p.i.), and the reduction rate of recovered larvae were 99.6% (FBZ) and 80.8% (ABZ) respectively. FBZ was more effective against early encapsulated larval T. spiralis (at 21 to 25 days p.i.), than ABZ when both were given at 20 mg/kg for five consecutive days (99.8% and 45.4% reduction, respectively). In conclusion, flubendazole was more effective than albendazole against adult and parenteral stages of Trichinella spiralis in mice. PMID- 11484355 TI - Experimental trichinellosis in fallow-deer (Dama dama L.). AB - Herbivora can play a very important role in spreading trichinellosis, as showed by the massive epidemics in man, caused by the consumption of horse meat in the last years. In this context, the present study has been undertaken to verify, through an experimental infection, the susceptibility, together with other biological parameters, of fallow-deer to Trichinella infection. The four animals, 8-9 months of age and 18-25 Kg body weight, were orally infected with low doses of Trichinella britovi and T. pseudospiralis (2,000 larvae/animal). After day 30 p.i., the animals were necropsied and, using artificial digestion methods, larval burden of Trichinella in muscle tissues was determined. Histopathological, serological (IgG monoclonal blocking ELISA) and biochemical data were assessed during the experiment. The results showed the susceptibility of fallow-deer to T. britovi and T. pseudospiralis infection; under the same inoculum size, the number of larvae/g was higher in group infected with T. britovi. The animals showed a higher immunological response to T. pseudospiralis infection. The results are discussed. PMID- 11484356 TI - Trichinella spp. in ostrich meat: a public health risk? AB - In the present work the biological behaviour of T. spiralis and T. pseudospiralis in ostriches is reported. Oral infections were performed in eight ostriches with two infective doses (10,000 and 80,000 larvae) for each species of Trichinella. On day 0, 30 and 60 p.i. blood samples were collected to assay the serum changes concerning specific muscle enzyme activities and total proteins. The immunological study, to determine specific IgG in sera, was conducted employing a monoclonal blocking ELISA. From the carcasses of sacrificed animals, samples of various muscle tissues were examined by the digestion method and by standard histopathologic procedures. The study showed a low susceptibility of the ostriches to T. pseudospiralis; preferential sites of larval distribution were muscle tissues of the legs. T. spiralis could be found in muscle tissues only when a high number of larvae were inoculated. Immunological reactivity was found only in animals infected with higher doses of T. pseudospiralis. PMID- 11484357 TI - Susceptibility of nutria (Myocastor coypus) to Trichinella infection: biological aspects. AB - Experimental infections with three different species of Trichinella in nutria in order to evaluate the susceptibility and the role of these rodents in the spreading of parasitosis in nature were carried out. The nutria is present in many italian wet areas and its distribution is expanding. The nutria meat is utilized as food in different countries and is retained responsible for trichinellosis in man. Two groups of ten animals were infected per os with 500 and 5,000 (n. 10) infective larvae of T. britovi; an additional study was arranged with two groups of animals infected with 5,000 larvae of T. spiralis and T. pseudospiralis, respectively. After 45 days, all animals were slaughtered and samples of different muscles were processed by standard artificial digestion and by routine histological methods. Serological investigations (specific IgG) have been carried out on sera samples by employing a monoclonal blocking ELISA. The animals showed a significant susceptibility to the infection with all species of tested Trichinella and immunological reactivity. Data obtained are discussed. PMID- 11484358 TI - Infectivity of Trichinella spp. recovered from decaying mouse and fox muscle tissue. AB - The tolerance to degradation processes in meat of nine Trichinella genotypes was studied in mouse and fox tissue, respectively. Minced muscle tissue with Trichinella larvae of different age was stored at room temperature at 100% relative humidity. During storage weekly sub samples of the minced meat were digested and released larvae were inoculated in mice to evaluate the Reproductive Capacity Index (RCI). The RCI decreased with the length of storage, but the larvae from older infections appeared better adapted to tolerate the degradation processes. The African species T. nelsoni had a relative higher tolerance to elevated temperature during storage and the unencysted species T. pseudospiralis was the most vulnerable genotype. PMID- 11484359 TI - The relevance of evolutionary genetics for identification of Trichinella sp. and other pathogens at the strain, subspecies and species levels. AB - In the specific field of Trichinella research, the impact of evolutionary methods has been until now limited. This is all the more distressing, since this group of parasites is characterized by hard taxonomical problems. Moreover, for the epidemiological tracking of Trichinella populations, high-discriminating genetic typing, conveniently analyzed by population genetic methods, could be of considerable help. With no additional efforts, many molecular data are already available and could be used for retrospective analyses able to solve many taxonomical problems. It is desirable that coming studies are specifically designed according to evolutionary genetics principles and the question under study (sample size, selection of the appropriate marker). PMID- 11484360 TI - Infectivity of Trichinella spiralis larvae in pork buried in the ground. AB - Time of survival and infectivity of Trichinella spiralis larvae in pig muscle tissue, buried at various depths in the ground were assessed. In the pork pieces the number of infective larvae was 250 ML/g. Meat originated from pig halves was divided in 39 equal pieces, 0.7 kg each, disposed in three groups of 13, and buried in depths of 30, 50, and 100 centimeters respectively. The pork was dug up at 13 intervals, approximately every week, until 91st day of the experiment. After each time interval, infectivity of larvae was assessed by bioassay on rats. The artificially infected rats were sacrificed on 42nd day after the infection and meat was examined by the following methods--artificial digestion and trichinoscopy. It was found that the larvae during all 90 days preserved infectivity in each depth. PMID- 11484361 TI - Infectivity, persistence and serological response of nine Trichinella genotypes in rats. AB - Domestic and sylvatic Trichinella genotypes were evaluated for infectivity, muscle larvae persistence, and host antibody responses in rats. Groups of rats were inoculated with T. spiralis, T. nativa, T. britovi, three genotypes of T. pseudospiralis (from USSR, USA, and Australia), T. murrelli, Trichinella T6, and T. nelsoni, respectively. The muscle larvae intensity (larvae per gram), total larval burden (lpg x rat weight), and the antibody levels were determined at necropsy 5, 10, 20, and 40 weeks post inoculation. All Trichinella genotypes were established in the rats, but infectivity and persistence differed significantly: T. spiralis established and persisted in high numbers, the three T. pseudospiralis genotypes were also highly infective but differed significantly in persistence, T. britovi and T. nativa had limited infectivity and persistence, Trichinella T6 had low infectivity and very low persistence, and T. murrelli and T. nelsoni were almost non-infective. Except for T. spiralis, initial total muscle larval burdens declined significantly for other genotypes during the experiment. A high initial serological response was detected for all genotypes, but the antibody levels decreased rapidly in relation to decreasing larval burdens. After 20 w.p.i. the antibody levels remained high only in T. spiralis and T. pseudospiralis infected rats. The high infectivity and persistence of T. pseudospiralis in rats, suggests that in addition to T. spiralis, this species might be of significant importance in the domestic cycle of trichinellosis. PMID- 11484362 TI - Reduction of muscle larvae burden in rats experimentally infected with Trichinella spiralis. AB - In Wistar rats infected with 500 to 2,500 Trichinella spiralis larvae the muscle larvae intensity (larvae per gram-l.p.g.) was measured from 20 to 180 day post infection (d.p.i). The l.p.g. increased to day 40-50 p.i. and decreased thereafter. The highest reduction took place between 60 and 120 d.p.i. with intermediate inoculum of T. spiralis larvae. The mechanism of the reduction of T. spiralis larvae in muscles is suggested to depend on pericapsular-intercapsular host cells infiltrations attracted by parasite antigens. PMID- 11484363 TI - Effect of viable or dead Lactobacillus casei organisms administered orally to mice on resistance against Trichinella spiralis infection. AB - The capacity of viable, dead Lactobacillus casei and supernatant from L. casei culture, administered by oral route, to induce resistance in mice against Trichinella spiralis infection was evaluated. The percentage of adult worm reduction in the intestine five days after T. spiralis infection as compared with the worm burden in the control group fluctuated between between 53.1 and 58% in mice treated with viable L. casei, while reductions in animals treated with dead lactobacilli or supernatant from L. casei culture were of 44 and 32.5% respectively. The percentage of larvae per gram of muscle tissue reductions, as compared with controls, obtained 30 days after infection ranged from 48.4 to 70.7% in rodents which ingested viable L. casei compared with the percentage reductions of 65.9 and 24% obtained respectively in mice treated with dead lactobacilli or with L. casei supernatant. The protective response observed in the present study may be explained on the basis of 1) lactobacilli colonization of the intestine, 2) macrophage processing of dead lactobacilli in local immune tissues and presentation of L. casei antigens to Th1 cells which, in turn, produced IL-2 to activate B cells and other T cells. PMID- 11484364 TI - Early detection of Trichinella spiralis infection by the polymerase chain reaction in blood samples of experimentally infected mice. AB - Up to now, there is no useful method to diagnose trichinellosis at the early stages of infection. The aim of the present investigation was to know if the polymerase chain reaction (PCR), could detect DNA of migratory larvae in mice experimentally infected with T. spiralis. Thirty-three Balb/c female mice 4.6 weeks old, were infected with 300 larvae of T. spiralis/mouse. Blood samples of these animals were positive for T. spiralis at 3rd and 17th post-infection days. The control group was negative. PCR could detect from one to 200 larvae. Our results showed that it was possible to make early diagnosis of trichinellosis in blood of mice infected with T. spiralis. PMID- 11484365 TI - Verification of good production practices that reduce the risk of exposure of pigs to Trichinella. AB - Control of Trichinella infection in swine has traditionally been accomplished by inspection of individual carcasses or by post-slaughter processing to inactivate parasites. Recent declines in prevalence of this parasite in domestic swine, coupled with improvements in swine management systems, offer the opportunity to document pork safety during the production phase. We report here on a certification pilot study using an audit to document good production practices for swine relative to the risk of exposure to trichinae. Based on the results, improvements in the program have been made and further studies will be undertaken prior to launching a voluntary trichinae herd certification program in the United States. PMID- 11484367 TI - A single, multiplex PCR for differentiating all species of Trichinella. AB - The genus Trichinella is currently divided into seven species and at least three additional, unclassified genotypes, Trichinella T6, T8 and T9, where both T8 and T9 have been deemed very similar to T. britovi. Other than for the non encapsulated species, the absence of distinguishing morphological characters and the overlapping nature of the biological characters within this genus make these traits unsuitable for diagnosis. Consequently, we have developed a simple PCR test for the unequivocal differentiation of all currently recognized species of Trichinella including Trichinella T6. DNA sequence data from each Trichinella genotype were generated from internal transcribed spacers, ITS1 and ITS2, and from expansion segment V (ESV) of the rDNA repeat, from which five different PCR primer sets were chosen. When used simultaneously, this primer mix generates a simple and unique electrophoretic DNA banding pattern for each species and genotype. The ESV-derived primer set contributes at least one band to each agarose gel-derived genotypic pattern and therefore functions as an internal control for PCR integrity. Geographical isolates of each Trichinella genotype were used to verify the reliability and reproducibility of respective DNA banding patterns using single muscle larvae. PMID- 11484366 TI - Epidemiological surveillance and control of trichinellosis in Buenos Aires province. AB - The Ministries of Health and Agrarian Concerns created in 1994 the "Zoonosis Commission", a working group of professionals of both institutions, for improving the epidemiological surveillance and control of trichinellosis in Buenos Aires Province. The main purpose was the early detection and control of swine trichinellosis, for reducing human outbreaks, the strategy of action was to decentralize epidemiological surveillance and control activities at municipal level, integrating 101 counties of the endemic area. Since 1996 a high number of infected pigs were detected previously to human consumption (years 1996-1999: 1,073 infected pigs), and since 1997 the occurrence of human outbreaks and swine foci have decreased. PMID- 11484368 TI - The lateral flow card test: an alternative method for the detection of Trichinella infection in swine. AB - A novel lateral flow card (TS-Card pork) test was developed for the serological detection of Trichinella infected pigs. Based on extensive studies performed in Romania during 1999-2000 this test proved to be highly specific sensitive, rapid (3-12 minutes) and easy to use (no need for laboratory facilities). It can be used both for the detection of Trichinella infection in carcasses and for epizooliological studies using a variety of samples including whole or dried blood, serum, or tissue fluids. The TS-Card pork test, used as a screening test, can be the foundation of an on-farm or field based inspection system to significantly improve food safety in countries with a high prevalence of Trichinella in pigs or other food animal species. The results presented are also promising for application of the test in an on-line laboratory based inspection system since the speed of the test allows sufficient time to rail out suspected hog carcasses during the slaughter process. PMID- 11484369 TI - Trichinellosis in farmed wild boar: meat inspection findings and seroprevalence. AB - A reflection of highly prevalent endemic wildlife trichinellosis is seen in wild boar farming in Finland. During the last five years, 0.7% (15/2265) of wild boars undergoing official meat inspection have been determined to be Trichinella positive. These findings originate from six different farms. In Finland, T. spiralis and T. pseudospiralis have been discovered in meat inspection of wild boars. ELISA showed 11 out of 99 serum samples (11%) as having specific antibodies for T. spiralis crude antigen. Positive samples were from three out of the thirteen farms from which the sera were available. Most of the positive serum samples (8/11) originated from a farm where trichinellosis was also revealed in meat inspection, the other two seropositive farms were without previous Trichinella records. Over the last few decades, no reports have been made of human trichinellosis acquired in Finland. This indicates both efficient meat inspection as well as public awareness of high-risk foodstuff. PMID- 11484370 TI - Localization of Trichinella spiralis in muscles of commercial and parasitologic interest in pork. AB - Trichinellosis is widespread around the world with different representatives of the genus Trichinella found in almost every continent. In Argentina the main source of transmission for the disease to humans is pig meat infected with Trichinella spiralis. The object of this work was to determine the distribution of Trichinella larvae in fresh meat cuts which are sold for human consumption and in the muscles traditionally used for the disease diagnosis at meat-packing plants. Cranial muscles to the last rib showed more Trichinella spiralis larvae than those with a caudal location (p < 0.01). No significant differences were found (p > 0.05) between bilateral left and right muscles. Significant larval concentrations were found in the neck muscles, even in carcasses with a low parasitic load; these muscles are used to prepare cold meats (boston butt). Commercial cuts of meat had a substantial larval burdens in animals experimentally infected with 500 to 5,000 Trichinella spiralis larvae, with parasite burdens similar to infection levels in muscles evaluated at the meat packing plant. PMID- 11484371 TI - Swine trichinellosis in slaughterhouses of the metropolitan area of Toluca. AB - In order to determine the prevalence of Trichinella spiralis infections in abattoirs of the metropolitan area of Toluca where pigs from commercial farms as well as backyard pigs are slaughtered, 539 swine diaphragm tissue samples were collected and examined by trichinoscopy and artificial digestion. Serum samples from the same animals were analyzed by ELISA using somatic and excretory/secretory antigens, and by Western blot analysis. T. spiralis muscle larvae were not found by trichinoscopy or artificial digestion. However, specific antibodies were detected by ELISA and confirmed by Western blotting in 12.4% of the serum samples examined. Analysis of risk factors showed no association of seropositive results with sex. However, significant higher risk was observed in swine seven to 12 months old and in backyard pigs, compared with pigs from commercial farms. PMID- 11484372 TI - A challenge of veterinary public health in the European Union: human trichinellosis due to horse meat consumption. AB - Human trichinellosis in the European Union due to insufficiently cooked horse meat consumption has been reported in France and in Italy during the past 25 years. It occurred in several outbreaks totalling more than 3,000 patients during this period, with a low mortality and a high morbidity. Causative Trichinella species or phenotypes were determined by the International Reference Laboratory for Trichinellosis in Rome, Italy. They were: Trichinella spiralis, T. britovi and T. murelli. As the culinary habits and customs of populations cannot be changed by regulations, measures of protection of public health essentially depend on food inspection. Comprehensive studies having been conducted in the pathophysiology of Trichinella infection in horses; it was demonstrated that the localisation of larvae are quite different in horses and in pork. It resulted an instruction from the French Veterinary, Service recommending that: the sampling of muscles in horses carcasses has to be done at first in the tongue (apex), then in the diaphragm (pillars); at least 50 g have to be sampled in each site; examination for larvae has to be done with the digestion method. Such recommendations might be extended to other EU member countries then to the OIE Zoo-Sanitary Code. PMID- 11484373 TI - Detection of Trichinella infection in slaughter horses by artificial digestion, ELISA and PCR. AB - In this study we compared the sensitivity of molecular, serologic and parasitologic methods for diagnosis of equine trichinellosis in two abattoirs, one rural and one federal inspection type. Diaphragm muscle samples were obtained from 170 slaughter horses and examined by artificial digestion and PCR. Serum samples from these horses were also analyzed by ELISA. No Trichinella muscle larvae were detected by artificial digestion. However, specific antibodies against Trichinella were detected in 17% and 7% of the serum samples examined from the rural and the federal abattoirs respectively. By PCR, 15% and 2% of the samples from these two abattoirs were Trichinella positive. PMID- 11484374 TI - Immunodiagnosis of Trichinella infection in the horse. AB - From 1998 to 2000, 5,267 horse sera were collected from several Trichinella regions in Romania. Sera were initially screened in laboratories in Romania, Serbia and Italy with an ELISA and a Western blot (Wb) using an excretory/secretory (ES) antigen and several conjugates (protein A, protein G, and sheep or goat anti-horse). Differences in serology results were obtained among the different conjugates and also between ELISA and Wb. Depending on the test used, specific antibodies were found at a prevalence rate of 3-6% of horses. Serum samples classified as positive were tested again by ELISA using a synthetic tyvelose glycan-BSA antigen, in Italy. All serum samples tested using this antigen were negative; in contrast, serum samples from experimentally infected horses were positive with the glycan antigen. The negative results obtained with the glycan antigen are consistent with the low prevalence of horse trichinellosis reported in the literature. Based on these results, further studies are needed to validate immunodiagnostic tests to detect Trichinella infection in horses. PMID- 11484375 TI - Horse trichinellosis, an unresolved puzzle. AB - In spite of routine controls to detect Trichinella larvae in horse-meat, human infections due to horse-meat consumption continue to occur in France and Italy. The epidemiology of horse trichinellosis since its discovery in 1975 is outlined, addressing the possible modes of natural transmission to horses, the need to develop more sensitive methods for detecting Trichinella larvae in horses, and the economic impact of horse trichinellosis. Investigations of human outbreaks due to horse-meat consumption have implicated single cases of inadequate veterinary controls on horses imported from non-European Union countries. In particular, most cases of human infection have been attributed to horses imported from Eastern Europe, where pig trichinellosis is re-emerging and the main source of infection in horses. PMID- 11484376 TI - The detection of encapsulated and non-encapsulated species of Trichinella suggests the existence of two evolutive lines in the genus. AB - In recent years, the discovery of many non-encapsulated isolates of Trichinella, designated Trichinella pseudospiralis and the identification of a new non encapsulated species, Trichinella papuae, has revealed that the biomass of the genus Trichinella does not only include the well known encapsulated species (T. spiralis, T. nativa, T. britovi, T. murrelli, and T. nelsoni) but also includes geographically disseminated, non-encapsulated species that represent important biological entities in the genus. Larvae of the first stage (L1) of both non encapsulated and encapsulated species are able to penetrate the muscle cell and induce a dedifferentiation of this cell. But following this point in the parenteral cycle, non-encapsulated and encapsulated species diverge with respect to their developmental strategies where L1 of encapsulated species are able to induce the nurse cell to synthesize collagen, unlike non-encapsulated larvae which do not induce collagen production. The presence or absence of a collagen capsule is of great importance in the natural cycle of these parasites in that it allows the encapsulated larva to survive to substantially longer periods of time and therefore remain infective even within putrified muscle tissue. PMID- 11484377 TI - Estimating the genetic divergence and identification of three Trichinella species by isoenzyme analysis. AB - Isoenzyme-based approach was applied to compare Trichinella spiralis, T. britovi and T. pseudospiralis species. Among 13 enzyme systems examined, esterase (EST), malic enzyme (ME) and phosphoglucomutase (PGM) have been found as fully diagnostic, with no common allele in species studied. Adenosine deaminase (ADA), adenylate kinase (AK), hexokinase (HK), peptidase leucyl-alanine (PEP-C) and fructose-bis-phosphatase (FBP) have been capable of distinguishing the two species from resulting profiles. In addition, ADA, AK and PGM displayed the enzyme expression in the lowest amounts of muscle larvae in systems tested (100 larvae/100 microliters of extracts). Based on allozyme data, T. pseudospiralis has been found as the most distinct species within the group of taxa. Only a subtle genetic variability was recorded for T. pseudospiralis in which solely phosphoglucomutase exhibited variant patterns. In addition to the study of reference isolates, T. spiralis from lowland fox in Eastern Slovakia has been evidenced by use of genetic markers. This finding has proved that T. britovi is not the exclusive species parasitizing in the sylvatic ecosystem of the Slovak region. PMID- 11484378 TI - Biological and genetic characteristics of two Trichinella isolates in China; comparison with European species. AB - The most recent taxonomic revision in Trichinella genus included 10 taxa. Trichinella spiralis (Ts) is a thoroughly studied species but most of the isolates came from the Europe or American continents. Few information is available from China about the Trichinella isolates and their diversity. In this report two Chinese isolates were characterized and compared with European strains. The in vitro release of newborn larvae (NBL) was determined for the two species. NBL is observed in supernatant of cell culture after adults purification on day 4 post infection (pi) with Ts, and on day 5 pi with Trichinella nativa (Tna). A new parameter was thus proposed to characterize Trichinella strains. Restriction fragment length polymorphism (RFLP) of genomic DNA and random amplified polymorphic cDNA (RAPD) were used to define genetic variability among different isolates. Species specific pattern could be obtained with RAPD but it was far more difficult to get geographical markers for Trichinella using these methods. A low genetic variability in Trichinella species (i.e. a strong "clonality") is suggested. PMID- 11484379 TI - Evaluation of two PCR-based techniques for molecular epidemiology in Finland, a high-endemic area with four sympatric Trichinella species. AB - Trichinella larvae collected from wildlife, domestic and synanthropic animals in Finland were identified to species by two molecular techniques: Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) and the recently described multiplex PCR. The RAPD-PCR was very sensitive to the sub-optimal preservation muscle larvae and resulting in weak and smeared bands on the gels for such material. However, the same samples yielded easily recognizable bands in the multiplex PCR; this latter technique is then recommended for epidemiological studies, especially when the preservation of the samples is sub-optimal. For larvae in good condition the unequivocal bands obtained by multiplex was the easiest identifiable. Four species of Trichinella were identified in the material: T. spiralis, T. nativa, T. britovi, and T. pseudospiralis. Trichinella britovi is a new record for Finland, and T. pseudospiralis is a new record for Northern Europe. Mixed infections between T. britovi and T. spiralis, T. nativa and T. spiralis, and between T. britovi and T. nativa were detected; this is the first record of a mixed infection between T. spiralis and T. nativa in a naturally infected host. Raccoon dogs were the only host species from which all of the four Trichinella species were detected. Trichinella spiralis was found in both domestic animals and wildlife, but none of the sylvatic Trichinella species were detected in domestic pig. PMID- 11484380 TI - Twelve years of activity of the International Trichinella Reference Centre. AB - The ITRC is the official reference laboratory of both the International Commission on Trichinellosis (since 1988) and the International Office of Epizootics (since 1992). The ITRC was created as a repository for Trichinella strains and as a source of materials and information for international research in 1988. To date, about 900 isolates of human and animal origin from throughout the world have been examined and identified by new procedures developed at the ITRC or in collaboration with other institutions. Using material from this collection, the ITRC has provided a complete revision of the systematics of the genus Trichinella. The ITRC database can be consulted by accessing the web-site: www.simi.iss.it/trichinella/index.htm. PMID- 11484381 TI - Invasion of epithelial cells by Trichinella spiralis: in vitro observations. AB - It has been known for many years that Trichinella spiralis initiates infection by penetrating the columnar epithelium of the small intestine, however, the mechanisms used by the parasite in the establishment of its intramulticellular niche in the intestine are unknown. The recent demonstration that invasion also occurs in vitro when infective larvae of T. spiralis are inoculated onto cultures of epithelial cells provides a model that allows the direct observation of the process by which the parasite recognizes, invades and migrates within the epithelium. The finding that penetration of the cell membrane or induction of plasma membrane wounds by larvae do not always result in invasion argue in favor of some kind of host-parasite communication in successful invasion. In this sense, the in vitro model of invasion provides a readily manipulated and controlled system to investigate both parasite, and host cell requirements for invasion. PMID- 11484382 TI - Expression of MyoD and myogenin in muscles of mice experimentally infected with Trichinella spiralis or Trichinella pseudospiralis. AB - We developed a detection system for myogenic regulatory factors such as MyoD and myogenin. Adapting the method we performed a longitudinal analysis of such regulatory factors after infection with T. spiralis and T. pseudospiralis. MyoD and myogenin were expressed from the early phase of cystogenesis in T. spiralis infection. The expression returned to the normal level after 18 days from the infection when the cyst was complete. In T. pseudospiralis infection, they were also expressed from the early phase of cystogenesis, but continuously expressed at least up to 43 days post infection. PMID- 11484383 TI - Ultrastructural characteristics of nurse cell-larva complex of four species of Trichinella in several hosts. AB - The nurse cell-larva complex of nematodes of the genus Trichinella plays an important role in the survival of the larva in decaying muscles, frequently favouring the transmission of the parasite in extreme environmental conditions. The ultrastructure of the nurse cell-larva complex in muscles from different hosts infected with T. nativa (a walrus and a polar bear), T. spiralis (horses and humans), T. pseudospiralis (a laboratory mouse) and T. papuae (a laboratory mouse) were examined. Analysis with transmission electron microscope showed that the typical nurse cell structure was present in all examined samples, irrespective of the species of larva, of the presence of a collagen capsule, of the age of infection and of the host species, suggesting that there exists a molecular mechanism that in the first stage of larva invasion is similar for encapsulated and non-encapsulated species. PMID- 11484384 TI - Detection of tyrosine phosphorylated proteins in Trichinella spiralis L1 larvae. AB - Western-blotting analysis showed the presence of tyrosine phosphorylated proteins in crude extracts of T. spiralis larvae and these phosphorylated proteins were located by immunofluorescence on the striations of the larval cuticle. The patterns of phosphorylated proteins were modified when larvae were incubated with bile. PMID- 11484385 TI - The epidemiology of human trichinellosis in China during 1964-1999. AB - The large foci of trichinellosis are mainly located in the southeastern, the central and northeastern China. By the end of 1999, human cases with trichinellosis have been recorded in 17 out of 34 Provinces/Autonomous Regions/Municipals (P/A/M) of China. The seroepidemiological surveys of T. spiralis infection in humans were carried out in nine out of 34 P/A/M. The overall seroprevalence was 5.3%. The prevalence detected by muscle biopsy in Henan province was 2.5%. From 1964 to 1999, 548 outbreaks of trichinellosis, with 23,004 cases and 236 deaths, were recorded in 12 P/A/M of China. Pork is the predominant source of outbreaks of human trichinellosis in China, 525 (95.8%) out of 548 outbreaks were caused by eating pork. However, 14 outbreaks resulting from the consumption of mutton and game meat occurred in recent years, suggesting the increasing significance of herbivorous and wild animals as source for human trichinellosis. PMID- 11484386 TI - Epidemiology of swine trichinellosis in China. AB - Swine trichinellosis has been reported in 26 provinces, autonomous regions or municipalities (P/A/M) of China. The prevalence rate in swine varied from 0.12% to 34.2% in five P/A/M, from 0.01% to 0.0001% in other P/A/M. The seroepidemiological survey of swine trichinellosis was carried out by ELISA in seven P/A/M, the seroprevalence varied from 0.09% to 29.63%. The prevalence of Trichinella infection in pork sold at the market was from 0.29% to 5.6% in four provinces. The transmission of trichinellosis by garbage is the main features of epidemiology of swine trichinellosis in China. Rat is an important reservoir in the domestic cycle of trichinellosis. The prevalence rates of T. spiralis infection in rats were from 1.98% to 15.06% in six provinces or autonomous regions. The treatment prophylaxis with forage contained albendazole has been applied to the control of swine trichinellosis in Nanyang area of Henan province, the prevalence of swine trichinellosis had decreased from 32.2% before prophylaxis to 0.12% after prophylaxis. PMID- 11484387 TI - Proceedings of the Tenth International Conference on Trichinellosis. August 20 24, 2000, Fontainebleau, France. AB - The Tenth International Conference on trichinellosis, organized by the International Commission on Trichinellosis, gathered together 147 scientists from 34 countries. The 173 presentations ranged over most aspects of this parasitic zoonosis: history, epidemiology, speciation, biology, immunology, biochemistry, human and animal pathology, new diagnostic methods and control. PMID- 11484389 TI - Outbreaks of human trichinellosis caused by consumption of dog meat in China. AB - Dog meat has become an important source of Trichinella infection for humans in China. The first documented outbreak of human trichinellosis resulting from the consumption of dog meat occurred in China in 1974. Until 1999, the outbreaks with this source of infection occurred mostly in Northeast of China (81 cases in five outbreaks in Jilin and two in Liaoning), Beijing (six cases) and Henan provinces (two cases). The epidemiological surveys were performed in nine Provinces or Autonomous Regions of China among 19,662 dogs samples. Dogs trichinellosis prevalence ranged from 7% in Henan to 39.5% in Heilongjiang, with an overall prevalence of 21.1%. Based on random amplified polymorphic DNA fingerprint (RAPD), some dog isolates of Heilonjiang and Jilin provinces were recently identified as Trichinella nativa, suggesting that this parasite is widely distributed among dogs in Northeast of China, while Trichinella spiralis in swine appears to be a common parasite throughout China. Since the Trichinella larvae in dog meat is resistant to freezing, caution should be paid to the consumption of dog meat even if it had been frozen. PMID- 11484388 TI - Epidemiological survey of Trichinella infection in some areas of Henan province. AB - During 1996-1998 epidemiological survey of domestic Trichinella spiralis infection and serological survey on human beings were carried out in Yanling, Xuchang counties and Luohe city. By the artificial digestion method, the positive rates of pork in market and mutton were 1.6% and 0.6% respectively. Meat samples of pigs from pig-farms, cats and dogs were all negative for Trichinella larvae. Trichinellosis infection levels in rats captured in peasant households and places around abattoirs were 0.5% and 4.1% respectively (P < 0.05). By ELISA (double antibody) with excretion-secretion (ES) antigens of Trichinella larvae, the positive rates of antibodies against Trichinella in sera of peasants and professional drivers were 2.2% and 7.8% (P < 0.05). The positive rates of antibodies in pigs from pig-farms and from peasant households were 1% and 7% respectively (P < 0.05). The levels of anti-Trichinella antibodies in rats captured in peasant households and places close to abattoirs were 0% and 11.1% respectively. Trichinella spiralis larvae were found in the washing meat slop in abattoirs. PMID- 11484390 TI - Human trichinellosis in Argentina. Review of the casuistry registered from 1990 to 1999. AB - Trichinellosis is an endemic disease in Argentina. During the last decade (1990 to 1999) 5,217 human cases were notified to the National System of Epidemiological Surveillance from 18 provinces of 23 that integrate the country. However 91% of the cases notified were registered in only three provinces (Buenos Aires, Cordoba and Santa Fe) with a population of more than 50% of the inhabitants of Argentina. This data suggest the magnitude of trichinellosis as a public health problem, focalized in the central area of the country. PMID- 11484391 TI - Sylvatic trichinellosis in Texas. AB - There are no published reports of domestic or sylvatic trichinellosis in Texas. The aim of the present survey was to determine the presence of Trichinella species in selected representative species of potential wildlife reservoirs in southern Texas. In 1998-99, tongues of 211 wild mammals were collected in southern Texas: 154 coyotes (Canis latrans), three bobcats (Lynx rufus), 32 racoons (Procyon lotor), 13 opossum (Didelphis marsupialis), four ocelots (Leopardus pardalis) and five wild boars (Sus scrofa). Presence of Trichinella sp. larvae was investigated by artificial digestion and larvae of positive samples were identified at the species level by a multiple-polymerase chain reaction analysis. Nine (5.8%) coyotes had trichinellosis; in the muscles of seven of these coyotes, the larvae were identified as Trichinella murrelli. This is the first report of sylvatic trichinellosis in Texas. PMID- 11484392 TI - Trichinellosis in Greece: a review. AB - Trichinellosis which constitutes a public health problem in many countries seems to be of no importance on both pig industry and public health in Greece, where in spite of a law requiring mandatory use of trichinoscopy during meat inspection, muscle larvae have not been found in slaughtered pigs since 1957 in Thessaloniki and 1967 in Athens. Since its first recovery in 1946 and up to 1952, human trichinellosis has been found or suspected in 22 persons in the area of Athens and Thessaloniki. Moreover, in 1968, T. spiralis larvae were found incidentally in a human with laryngeal tumor and in 1971, living larvae were postmortem recovered in the diaphragm of a 70-year-old man. The average incidence of infection in pigs at that time was 0.02-2.2%. Since then, no other clinical case had been reported up to 1982-1984 when 15 people were found to harbor the parasite and fourteen of them were part of an outbreak which occurred in a small village in Northern Greece. Moreover, 1.07% of the pig serum samples which came from the same area, showed the presence of specific antibodies. Because of the above data, it is generally accepted that in Greece T. spiralis is only rarely spread in man. PMID- 11484393 TI - Epidemiological studies on animal and human trichinellosis in Estonia. AB - From 1992 to 1999, muscle samples from 814 sylvatic animals and 1,173 domestic and synanthropic animals were collected in 15 districts of Estonia; the prevalence of trichinellosis ranged from 1.0% to 79.4% for sylvatic animals and from 0.6% to 24.5% for domestic or synanthropic animals and for animals from fur bearing farms. The most important reservoirs of Trichinella in nature were the raccoon dog, the red fox, the lynx and the wolf. Three species of Trichinella (T. spiralis, T. nativa, and T. britovi) were identified by several types of PCR based analyses. Meat from sylvatic animals was the main source of Trichinella infection for humans. PMID- 11484394 TI - High prevalence of Trichinella nativa infection in wolf (Canis lupus) populations of Tvier and Smoliensk regions of European Russia. AB - Domestic and sylvatic trichinellosis have frequently been documented in European regions of Russia, with the highest prevalence reported in wolves (Canis lupus). From 1998 to 2000, 75 carcasses of wolves shot by hunters were tested for Trichinella larvae, and 73 (97.3%) of them were found to be positive. This very high prevalence of infection, the highest ever detected in a natural population of carnivores, could be explained by the human impact on the natural ecosystem. In fact, the diet of wolves living in the region under study mainly consists of carcasses of dogs and wolves, which are left in the forest or used as bait by hunters. PMID- 11484395 TI - Epidemiological studies on Trichinellosis among swine, wild boars and humans in Poland. AB - The introduction of the digestion method (the magnetic stirrer type) resulted directly in the reduction of Trichinella sp. infection among people in Poland. Pork and its products are still the main cause of human trichinellosis in Poland. However, epidemics caused by eating wild boar meat suggested that this way of the transmission of Trichinella sp. larvae to humans might be of considerable importance. PMID- 11484396 TI - Epidemiology of swine trichinellosis in the Republic of Croatia. AB - In order to study the epidemiology of trichinellosis with special emphasis on swine a detailed study was performed. The goals of the study were to define the prevalence of trichinellosis in swine and rats and to determine the etiological agents of the disease with special emphasis on sylvatic species, Trichinella britovi. Considering the obtained data it can be concluded that the risk of Trichinella infection for swine in endemic areas in Croatia is rather high today. On the contrary, a sharp decline in human cases is registered what is certainly a result of obligatory trichinelloscopy even for private purposes. The parasite was found to be prevalent in rats too. Studies on genotypes suggest that Trichinella spiralis is the most prevalent etiological agent of domestic trichinellosis in Croatia, even so the sylvatic species Trichinella britovi can be also present in the domestic habitat suggesting a link between the sylvatic and domestic cycle. PMID- 11484397 TI - Present status of trichinellosis in Yugoslavia: Serbia. AB - Trichinellosis was recognized almost one century ago as a health and animal husbandry problem in Serbia. In the last 10 years, trichinellosis has been expanding from three endemic regions (Srem, Macva and Negotinska Krajina) to neighbouring regions. The infection rate detected by veterinary inspection in 1999 year was 0.17% in slaughtered swines. Simultaneously, the number of infected humans increased three-five times in comparison with the period 1980-1990. For instance, 555 individuals were registered in 1999 as infected after the consummation of non-inspected pork from domestic swine or wild boar. Prevalence of trichinellosis in wild animals was examined more than a 20 years ago. The trichinellosis in horses has not been detected in the country, but infected horses imported from Serbia were detected in France and Italy. PMID- 11484398 TI - Trichinellosis in Romania: a short review over the past twenty years. AB - This paper emphasized the evolution of trichinellosis in man and animals during the last two decades in Romania. Between 1983 and 1993, human cases with Trichinella increased from 217 to 3,649. A similar evolution was observed in pigs. This situation was caused by the large spread of trichinellosis from big "industrial" farms. Complex chemoprophylaxis and treatment chemoprophylaxis gave some positive results to control trichinellosis in pig. PMID- 11484400 TI - [Evaluation of plain abdominal radiography prescriptions in a university hospital center]. AB - OBJECTIVES: The aim of this study was to evaluate the prescription, impact and diagnostic utility of plain abdominal radiography in a University Hospital. METHODS: Plain abdominal radiography series performed over 15 consecutive days were evaluated for quality and agreement with the patient's clinical status. Agreement between the final diagnosis and the radiographic findings were also compared. Contribution of plain abdominal radiography to final diagnosis was assessed on the basis of the conclusions on the hospital discharge report or from those drawn from a telephone survey. The same relationship was also assessed in terms of clinician status (medical student, resident physician, senior physician). RESULTS: The series included 175 radiography series (11.6 per day). In 86% of the cases, the choice of views was adapted to the clinical context. In 14% it was not. The quality of the x-rays was found to be unsatisfactory in 17%, good in 63% and excellent in 20%. The radiography was considered to be contributive to the final diagnosis in 13% of the cases and non-contributive in 87%. Clinician status was not correlated with the degree of usefulness for final diagnosis. Finally, the prescription was in agreement with standard guidelines in 28% of the cases and not in agreement in 72%. CONCLUSION: Plain abdominal radiographs are neither sensitive nor specific, frequently misleading, and costly per specific and correct diagnosis. Better physician awareness is required to limit the number of unnecessary examinations. PMID- 11484399 TI - [Procalcitonin in pyelonephritis and acute community-acquired pneumonia in adults]. AB - OBJECTIVES: Procalcitonin (PCT) is an acute-phase protein involved in the specific inflammatory reaction to severe bacterial or fungal infections. This protein does however lack sensitivity in focal infections. PATIENTS AND METHODS: In this study, we investigated PCT, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), leukocytosis, and fibrinogen levels at admission among adult patients hospitalized for community-acquired pneumonia (n = 33) or pyelonephritis (n = 30) and in a control population (n = 27) of patients with viral infections and non-infectious inflammatory diseases. RESULTS: Median serum PCT in the control group (0.21 ng/ml) was significantly lower than in the pyelonephritis group (0.46 ng/ml, p < 0.0005) or the pneumonia group (0.88 ng/ml, p < 0.0005). In the control group, median CRP was 51.4 ng/l reaching 220 mg/l in the pyelonephritis and 198 mg/l in the pneumonia group (p < 0.0005 in both cases). The other markers of inflammation investigated (leukocytosis, ESR, fibrinogen) did not show such differences between the control group and the sepsis groups. The sensitivity of PCT (threshold 0.5 ng/ml) was 61% for the diagnosis of pneumonia and 44% for the diagnosis of pyelonephritis. Specificity was 92% in both cases. In comparison, the sensitivity of CRP (threshold 50 mg/l) was 94% and 91% for pyelonephritis and pneumonia respectively with a 33% specificity in both cases. CONCLUSION: PCT is a specific but poorly sensitive marker of community-acquired pneumonia and pyelonephritis among adults hospitalized in medical wards. PMID- 11484402 TI - [Acute pancreatitis after high-dose interferon therapy in a patient with melanoma]. PMID- 11484401 TI - [Diaphragm-like strictures of the colon from diclofenac]. AB - BACKGROUND: Diaphragm-like strictures of the colon induced by nonsteroidal antiinflammatory drugs (NSAID) constitute a recently described uncommon clinical entity. CASE REPORT: An 86-year-old man consulted for abdominal pain and constipation. The physical examination revealed bloody stools and anemia in a patient on long-standing diclofenac therapy. Colonoscopy demonstrated two diaphragm-like strictures of the left and transverse colon with ulceration of the apex. Clinical outcome was favorable after discontinuing the NSAID. DISCUSSION: We compared this observation with 26 other case reports in the literature. The etiological mechanism appears to be related to systemic and local toxicity. Severe stenotic stricture may require surgical management. Endoscopic pneumatic dilatation may be an alternative to surgery. PMID- 11484403 TI - [Lymphedema of the upper limb, a complication of rheumatoid polyarthritis]. PMID- 11484404 TI - [SUNCT syndrome sensitive to lamotrigine]. PMID- 11484405 TI - [Prescribing home-care hospitalization elaborating a therapeutic project]. PMID- 11484406 TI - [Social security financing and hospital budgets]. PMID- 11484407 TI - [Classification of vasculitis]. PMID- 11484408 TI - [Treatment of vasculitis with antineutrophil cytoplasmic autoantibodies (ACPN)]. AB - DIAGNOSIS OF RENAL INVOLVEMENT: The diagnosis of renal involvement in patients with Wegener's disease and microscopic polyangeitis is facilitated with the search for antineutrophil cytoplasmic autoantibodies (ANCA) but still requires biopsy for confirmation. TWO THERAPEUTIC PHASES: Induction should be initiated immediately, and generally leads to initial remission. Long-term treatment is then needed to reduce the risk of recurrence and relapse. FOLLOW-UP: Close monitoring of ANCA levels is important in these patients. Several protocols are currently under evaluation with the aim of limiting immunosuppression toxicity. PMID- 11484409 TI - [Treatment of vasculitis without antineutrophil cytoplasmic autoantibodies]. AB - RENAL INVOLVEMENT: Several types of vascularides without antineutrophil cytoplasmic autoantibodies (ANCA) can affect the kidney. The most frequently encountered are small-vessel vascularides leading to glomerulonephritis: Goodpasture's disease, lupus, and, most importantly, rheumatoid purpura and type II mixed cryoglobulinemia secondary to hepatitis C. Medium-sized vessel vascularitis, such as occurs in polyarthritis nodosa, is exceptional and leads to renal ischemia. Renal involvement in large-vessel vascularities is also exceptional, e.g. temporal artery arteritis and Takayasu's syndrome. THERAPY: Treatment must be adapted to the type of vascularides and also its class (small, medium, or large-vessel disease) and depends on the gravity of each individual patient's situation. A large range of treatments have been proposed, but there is little solid evidence concerning efficacy. PMID- 11484410 TI - Calcium-activated potassium current in single Novikoff cell. AB - The electrophysiological properties of the hemichannels of the gap junction in a single Novikoff cell was studied using patch-clamp whole-cell recording techniques. During the experiments, a transient outward current (peak current, Ip) was found to be evoked by the increment of depolarized pulses. Ip was voltage dependent and could be inhibited partly by CsCl and tetraethylammonium (TEA), but was blocked completely by quinidine. When the intracellular free Ca2+ concentration ([Ca2+]i) was increased, the amplitude of Ip was enhanced. Adding 10 mmol/l EGTA in the pipette solution, Ip disappeared. When the cells were bathed in symmetrical high-K+ solution, Ip also completely disappeared. These results suggest that Ip is a calcium-activated potassium current, although it cannot be blocked by apamin. PMID- 11484411 TI - In vitro activity of sertaconazole, fluconazole, ketoconazole, fenticonazole, clotrimazole and itraconazole against pathogenic vaginal yeast isolates. AB - The in vitro activity of sertaconazole was compared with those of the most commonly used vaginal antimycotic agents--fluconazole, ketoconazole, fenticonazole, clotrimazole and itraconazole--against 94 strains of clinical isolates of Candida spp. using a macrodilution method in Casitone agar medium. The sertaconazole concentration (microgram/ml), at which 90% of the strains were inhibited, was 0.06 for C. albicans, 0.25 for C. glabrata and C. parapsilosis, 1 for C. krusei and 2 for C. tropicalis. These values show that sertaconazole is one of the most active products against yeasts causing vulvovaginal candidiasis, its activity against C. glabrata being particularly relevant. PMID- 11484412 TI - Radical degradation of high molecular weight hyaluronan: inhibition of the reaction by ibuprofen enantiomers. AB - The antioxidative and/or free-radical-scavenging activities of R-(-)- and S-(+) ibuprofen enantiomers, as well as of the drug racemate, were studied in vitro on measuring the kinetics of (uninhibited or drug-inhibited) degradation of high molecular weight hyaluronan by hydroxyl radicals. The continual flux of OH radicals at aerobic conditions was maintained by the H2O2 + Cu2+ system. The kinetics of hyaluronan degradation was monitored indirectly by capillary viscometry. Under experimental conditions, with no drug addition, the relative viscosity ([eta]rel) decreased continuously, reaching 13% of the initial [eta]rel. value in 4 h. Each drug tested exhibited a dose-dependent protective effect against hyaluronan degradation, however R-(-)-ibuprofen demonstrated a slightly greater activity than the drug S-(+)-enantiomer. PMID- 11484413 TI - ATP-induced mucin release from cultured airway goblet cell involves, in part, activation of phospholipase A2. AB - Adenosine triphosphate (ATP) has been shown to stimulate mucin release by activation of protein kinase C (PKC) following activation of phospholipase C (PLC) coupled to the P2 receptor via G-proteins. The aim of the present study was to investigate pathways downstream to the PKC activation in ATP-induced mucin release from primary hamster tracheal surface epithelial (HTSE) cells. The release of mucin was determined by chromatographic procedure after metabolic labeling of mucin with [3H]-glucosamine. The results were: i) ATP induced the release of arachidonic acid, which caused the release of mucin. Pretreatment with mepacrine (0.3 mM), a phospholipase A2 (PLA2) inhibitor, inhibited the ATP induced arachidonic acid and mucin release. Oleoyloxyethylphosphocholine, another PLA2 inhibitor, gave similar results. ii) An activator of PKC, 4 beta-phorbol-12 alpha-myristate-13-acetate (PMA, 1 microM) induced mucin release, which was inhibited by mepacrine pretreatment. iii) Downregulation of PKC by prolonged (16 h) PMA treatment caused inhibition of ATP-induced mucin release. Treatment of PKC downregulated HTSE cells with mepacrine did not further decrease the ATP-induced mucin release. These results suggest that PLA2 is involved in ATP-induced mucin release and its activation is sequential to the PLC-PKC pathway. PMID- 11484414 TI - Effect of astaxanthin on the hepatotoxicity, lipid peroxidation and antioxidative enzymes in the liver of CCl4-treated rats. AB - Astaxanthin is one of many carotenoids present in marine animals, vegetables and fruits. Since carotenoids are known to have antioxidant properties, we tested to determine if astaxanthin could have protective effects in the CCl4-treated rat liver by activating the antioxidant system. Astaxanthin blocked the increase of glutamate-oxalacetate transaminase (GOT) and glutamate-pyruvate transaminase (GTP) activity and thiobarbituric acid reactive substances (TBARS) in response to carbon tetrachloride (CCl4), while causing an increase in glutathione (GSH) levels and superoxide dismutase (SOD) activities in the CCl4-treated rat liver. These results suggest that astaxanthin protects liver damage induced by CCl4 by inhibiting lipid peroxidation and stimulating the cellular antioxidant system. PMID- 11484415 TI - Effect of duration of pilocarpine-induced status epilepticus on subsequent cognitive function in rats. AB - The aim of this study was to determine the effect of the duration of pilocarpine induced status epilepticus (SE) on subsequent cognitive function in rats. SE was induced by pilocarpine (320 mg/kg i.p.) and was terminated by injection of 1 mg/kg diazepam at 30, 60 and 90 min in 3 groups of 10 rats each. Cognitive function was tested by a passive avoidance task and was assessed at the baseline and on days 1, 7, 14 and 21 (post SE). It was found that cognitive function was disrupted on days 7, 14 and 21 post SE in rats who had SE for 60 and 90 min, whereas it was not affected in rats that had 30 min of SE. Hence, the duration of SE may affect future cognitive performance and mandates emergency treatment. PMID- 11484416 TI - Effects of bupivacaine and ropivacaine on hemodynamic parameters in rabbits. AB - In the present study the toxic effects of ropivacaine and bupivacaine on the cardiovascular and respiratory systems of rabbits were studied. Both drugs were administered intravenously at doses of 0.5, 1, 2.5, 5 and 10 mumol/kg. The effects of the two drugs on blood pressure, ECG and respiration rate were evaluated by considering the changes occurring 30 sec after intravenous bolus injection. High doses (5 and 10 mumol/kg) of bupivacaine and ropivacaine significantly reduced the heart rate and systolic pressure (p < 0.05). Five and 10 mumol/kg of bupivacaine significantly reduced diastolic pressure (p < 0.05), but only the 10 mumol/kg dose of ropivacaine had the same effect. Low doses of bupivacaine and high doses of ropivacaine significantly increased the PR interval, QRS duration and QT interval (p < 0.05). The 5 mumol/kg dose of bupivacaine caused ventricular tachycardia in 3 of 6 rabbits, whereas ropivacaine caused tachycardia in 1 of 5 rabbits. Neither drug had a significant effect on respiration rate or blood gas values (p > 0.05). The results indicate that ropivacaine is less cardiodepressive and arrhythmogenic than bupivacaine. PMID- 11484417 TI - Pharmacokinetic study of paclitaxel as a 3-hour infusion in an Indian population: 135 mg/m2 vs. 175 mg/m2. AB - The objective of this clinical study was to determine the pharmacokinetic parameters of paclitaxel at doses of 135 and 175 mg/m2 when given as a 3-hour infusion in an Indian population. Twelve cancer patients of both sexes participated in the study and a parallel experimental design was adopted for the assignment of doses to patients. A solid phase extraction technique was used for sample clean-up followed by a reversed phase HPLC assay for the analysis of paclitaxel in plasma samples. Pharmacokinetic parameters such as Cmax, AUC0 infinity, T1/2 beta, AUMC0-infinity, VSS, VZ and CLT were determined by a compartment model-independent method using a PCNONLIN package. Teff and AUCeff were also calculated and compared at the two doses by considering a plasma concentration of > or = 0.05 microM as threshold. The mean Cmax and AUC0-infinity values were 2.57 microM and 12.06 microM at the 135 mg/m2 dose level while at the 175 mg/m2 dose the values increased to 4.96 microM and 9.52 microM.h/l, respectively. It was found that the 135 mg/m2 dose resulted in greater mean CLT and VSS values than the 175 mg/m2 dose. The disposition of paclitaxel was found to be nonlinear and the pertinent pharmacokinetic parameters were comparable to those from previous clinical studies. It was concluded from the present study that further clinical trials of paclitaxel alone or in combination with other drugs should be undertaken cautiously, taking into consideration its nonlinear pharmacokinetics which necessitate proper adjustment of the infusion schedule and/or dose to avoid any adverse consequences to the patient. PMID- 11484418 TI - Effects of absorbed components of saiboku-to on the release of leukotrienes from polymorphonuclear leukocytes of patients with bronchial asthma. AB - Previous studies confirmed the efficacy of the Chinese herbal remedy Saiboku-to in patients with steroid-dependent asthma. We studied 8 phenolic compounds, isolated from the urine of patients receiving Saiboku-to, with respect to their effects on leukotriene (LT) release by polymorphonuclear leukocytes (PMN) obtained from 6 patients with atopic asthma and 8 healthy subjects. The compounds (0.01-10 micrograms/ml) were incubated with Ca2+ ionophore (A23187)-stimulated PMN, and concentrations of leukotriene B4 (LTB4) and leukotriene C4 (LTC4) in the supernatant were measured by EIA. Each compound suppressed the release of LTB4 and LTC4 by PMN obtained from healthy and asthmatic subjects. In particular, baicalein and magnolol were 5-10 times more potent than azelastine, an antiallergic drug, and significantly suppressed LTC4 release by PMN obtained from asthmatic patients, as compared with healthy subjects. Suppression of LTC4 release by these compounds may play an important role in the clinical efficacy of Saiboku-to. PMID- 11484419 TI - Lactic acidosis in the critically ill. PMID- 11484420 TI - Analysis of foodborne disease in Belgium in 1997. AB - Foodborne disease represents a major problem for public health in industrialized countries, albeit with a low lethality. Foodborne diseases are defined as a group of viral, bacterial or parasitic gastrointestinal infections transmitted by means of food. Proper food-hygiene practices and surveillance of individual diseases and in particular outbreaks are the first steps in targeting their prevention. The incidence of this illness is difficult to estimate. In the Netherlands a yearly incidence of gastrointestinal infections of 500 per 1,000 inhabitants is estimated, of which most are foodborne. To set up priorities in the actions to undertake, to establish the most frequent risks, to develop preventive efforts and to answer to international requirements, accurate data on foodborne disease from Belgium are required. In order to co-ordinate the initiatives in the Belgian context, a working group was set up in 1995. In 1997 a total of 2,013 persons with foodborne disease were identified as part of 140 outbreaks, 22 of which occurred with 10 cases or more. Salmonella Enteritidis (88 outbreaks) was identified as the main pathogen in foodborne disease, followed by S. Typhimurium (11), S. Hadar (4). Eggs and meat products were identified as the main food-items involved, although it remains difficult to obtain proper intervention studies allowing to identify the specific cause(s). In 1997, a total of 12,732 human Salmonella isolates and 5,617 Campylobacter isolates were identified by the respective national reference laboratories. Salmonella isolates from Belgium accounted in 1997 for more than a fifth of all Salmonella isolates in the EU. The final objective of the working group is the implementation of a surveillance system for all risk factors concerned with the development of food-related illness, including an early warning system and an efficient analysis of microbiological criteria relating to human health, food and food production, including livestock. An essential element of this surveillance is communication of the results, risks and measures for prevention between all the departments, institutions and public health authorities concerned. PMID- 11484421 TI - A Campylobacter coli foodborne outbreak in Belgium. AB - In May 1995, the Scientific Institute of Public Health was informed of an outbreak of gastrointestinal illness in a congregational school in the Brussels area. The field investigation identified 24 cases with mild to severe gastrointestinal and general symptoms of acute bacterial enterocolitis. Campylobacter coli was detected in the stools of 5 patients. A retrospective cohort study suggested that a mixed salad (containing ham and feta cheese) was the probable source of infection, but the route of contamination remained unknown. The rapid investigation of such episodes of collective foodborne infections is essential for the implementation of adequate control measures. PMID- 11484422 TI - In vitro study on the antimicrobial activity of various antibiotics against clinical isolates of Streptococcus pneumoniae from Belgium collected during winter 1998-1999. AB - A total of 205 isolates of Streptococcus pneumoniae obtained from 10 different centres were included in this study. The susceptibilities to penicillin, ampicillin, amoxicillin, amoxicillin/clavulanic acid, cefaclor, cefuroxime, cefotaxime, imipenem, ciprofloxacin, gemifloxacin, grepafloxacin, levofloxacin, trovafloxacin, erythromycin, clarithromycin, miocamycin, clindamycin and tetracycline were determined by a microdilution technique following NCCLS recommendations. Decreased susceptibility to penicillin was 16.1% [6.8% intermediate (0.12-1 microgram/mL) and 9.3% high-level (> or = 2 micrograms/mL)], cefotaxime insusceptibility (> or = 1 microgram/mL) 12.7%, ciprofloxacine insusceptibility (> or = 2 micrograms/mL) 15.6% with 1.5% of high level resistance (> or = 4 micrograms/mL), erythromycin insusceptibility (> or = 0.5 microgram/mL) 36.1% and tetracycline insusceptibility (> or = 4 micrograms/mL) 22.9%. Decreased susceptibility to cefotaxime was found in 78.8% of the penicillin-insusceptible isolates. No decreased susceptibility was found for gemifloxacin (> or = 0.5 microgram/mL) and trovafloxacin (> or = 1 microgram/mL). Compared to the 1996-1997 surveillance, penicillin, cefotaxime and erythromycin insusceptibility rose by 3.8%, 5.2% and 5.0% respectively, while tetracycline insusceptibility decreased with 8.2%. MICs of all beta-lactams rose with those of penicillin for penicillin-insusceptible isolates. Amoxicillin +/- clavulanate, cefotaxime and imipenem were generally 1, 1 and 5 doubling dilutions respectively more potent than penicillin on these isolates. Penicillin, ampicillin and cefuroxime were equally active while cefaclor was generally 5 dilutions less potent. Most penicillin-insusceptible isolates remained fully susceptible to amoxicillin +/- clavulanate and imipenem. The penicillin-insusceptible isolates were 36.4%, 27.3% and 3.0% co-insusceptible to erythromycin, erythromycin plus tetracycline and tetracycline respectively. A subpopulation of 52 isolates obtained from children aged < or = 3 years was also studied. Compared to the other isolates we found a statistically significant increase in insusceptibility for penicillin, cefaclor, cefuroxime, erythromycin, clarithromycin and tetracycline while a significant decrease was found for ciprofloxacin. PMID- 11484423 TI - Detection time of drugs of abuse in urine. AB - Estimating the detection time of a drug in urine is complex because of many different influencing factors and the lack of experimental data. Detection times vary depending on dose and route of administration, metabolism and characteristics of the screening and confirmation assays. Using a cut-off value of 1000 ng/mL, urinary samples can be positive for amphetamine for up to 5 days after intake of the drug. At the lower 300 ng/mL cut-off, amphetamine will be detectable one day longer. Very few data are available for designer amphetamines. After smoking one marijuana cigarette, THCCOOH (9-carboxy-delta 9 tetrahydrocannabinol) is detectable (using a screening cut-off of 50 ng/mL) for 2 4 days. More frequent use will be detectable for almost 1 month, exceptionally 3 months. Immunoassays to detect cocaine are targeted against the metabolite benzoylecgonine and use a cut-off of 300 ng/mL. An intravenous dose of 20 mg cocaine can be detected for 1.5 days. Street doses (administered via different routes) are detectable up to 1 week, and extremely high doses up to 3 weeks. Heroin rapidly metabolizes to 6-acetylmorphine and morphine. Immunoassays for heroin are calibrated with morphine but important cross-reactivity occurs and positive results must be confirmed by GC-MS. Experimental data for total morphine using a cut-off of 300 ng/mL suggest a detection time of 1 to 1.5 days for relatively low doses of heroin (3-12 mg) administered via i.v., IN or i.m. route. PMID- 11484424 TI - The hypereosinophilic syndrome after residence in a tropical country: report of 4 cases. AB - Severe eosinophilia may be complicated by acute or chronic visceral damage. The underlying origin of the hypereosinophilia may be infectious, allergic, toxic, malignant or systemic (the secondary or reactive hypereosinophilic syndrome), but in a number of cases no cause can be found (the idiopathic hypereosinophilic syndrome). We describe 4 cases with hypereosinophilia and secondary visceral damage after residence in a tropical region. In three cases a helminthic infection was the obvious cause, the brain and the heart were the target organs. After treatment of the infection both the hypereosinophilia and the neurological and cardiac lesions disappeared. The fourth patient died of multi-organ disease. No definite trigger of the hypereosinophilia could be found. We discuss clinical findings, necessary investigations and therapeutic strategies. PMID- 11484425 TI - Localised obliterative bronchitis due to non-occupational mineral dust inhalation. AB - We present a 56-year-old woman with an infiltrate in the right middle lobe secondary to obstruction of the right middle lobe bronchus by extensive submucosal fibrosis. This unique stenotic lesion of a central bronchus was caused by a domestically acquired localised central obstructive bronchitis due to the inhalation of mixed dust fibers, in the absence of advanced silicosis. This complication is a rare presentation of the myriad of pulmonary diseases associated with mineral dust inhalation. PMID- 11484426 TI - Report a case of Campylobacter coli bacteremia in a patient with gastro enteritis. PMID- 11484427 TI - [Psychiatric illness does not protect against a somatic disease]. AB - In four patients, two women aged 68 and 73 years and two men aged 65 and 57 years, serious diseases were not recognised because of more prominent psychiatric symptoms. Three of the patients had malignancies and one was suffering from dehydration with a urinary tract infection and de novo diabetes mellitus. The patient with a meningioma recovered after surgery, the other three patients died despite treatment. Somatic diseases may not be recognised in patients with psychiatric illness because doctors may be more likely to ascribe the symptoms to a psychiatric cause. Patients may also present their symptoms in an unusual way. It is important not only to diagnose the psychiatric disease but also to check the patient's physical condition. This applies particularly to elderly patients, because they are more likely to have somatic diseases. Good education of psychiatrists in terms of somatic diseases remains important. PMID- 11484428 TI - [Bisphosphonate treatment prevents hip fractures in 70-79 year old women with osteoporotic vertebral fractures]. AB - According to the data of a fracture intervention trial, in women aged 55-80 years with vertebral fractures or osteoporosis diagnosed by bone mineral density measurement, treatment with the bisphosphonate alendronate prevented hip fractures with numbers-needed-to-treat within 5 years of treatment of 46 and 66, respectively. In a large risedronate hip fracture study, this new bisphosphonate only showed a beneficial effect in women aged 70-79 years with moderately severe osteoporosis as judged by femoral neck T-score, when one or more vertebral fractures were present at the start of the treatment. The number-needed-to-treat was 29. However, in women aged over 80 years and who were selected predominantly on the basis of clinical risk factors for hip fracture, no effect was found with this drug on hip fracture rate, suggesting that most were not osteoporotic and/or that the clinical risk factors used did not have the clinical utility in identifying hip fracture risk. Other factors besides osteoporosis may play a more important role in causing hip fracture in this elderly group. Diagnosis of osteoporotic vertebral fractures in women aged 70-79 years is predictive of not only new vertebral fractures but also of hip fractures, and could therefore form an indication for drug treatment. PMID- 11484429 TI - [National hepatitis B vaccination closer to implementation, but not soon enough; recommendations from the Dutch Health Council]. AB - A Health Council Committee has advised the Minister of Health, Well-being and Sports not to implement universal hepatitis B vaccination in the Netherlands, as recommended by WHO and requested by members of the Dutch parliament. The Committee argued that the prevalence of hepatitis B carriers among the native Dutch population is so low (0.07%) that universal vaccination is not relevant. In contrast, vaccination was advised for children from parents who had immigrated from middle- and high-endemic countries to the Netherlands, because horizontal transmission is an important route of transmission among these people, as it is in their country of origin. This concerns an estimated 15% of the entire population. For the rest of the Dutch population the risk of horizontal transmission was considered negligible. Sexual transmission was considered more important. The optimal age of vaccination to prevent sexual transmission was considered to be shortly before puberty. The committee concludes that, as yet, no data is available to justify universal vaccination of this age group and highlights the need for additional studies, including those into the efficacy of existing preventive programs, before universal vaccination should be implemented. However, it can be argued that it is unlikely that sufficient information will be available for at least another five years, and it is certain that the risk of sexual transmission will increase during this time. Universal childhood vaccination is an investment in the youth which will pay out in the future, and which should therefore not be postponed. PMID- 11484430 TI - [Long-term use of benzodiazepines]. AB - Benzodiazepines are the most prescribed drugs in the Netherlands. There is scarcely an indication for long-term benzodiazepine use. Long-term use may lead to dependency and is associated with an increased risk of accidents/falls and cognitive function impairment. Therefore national and international guidelines advocate a conservative prescription policy, especially with respect to long-term prescription. It appears that these guidelines are not followed in practice. A standard sized general practice in the Netherlands contains, on average, 75 long term benzodiazepine users. There appear to be both patient and GP related factors which influence the maintenance of this long-term use of benzodiazepines. PMID- 11484431 TI - [Treatment methods for discontinuation of long-term benzodiazepine use]. AB - Treatment strategies for discontinuing long-term benzodiazepine usage can be divided into minimal interventions and gradual discontinuation programs. Minimal interventions invite patients to quit their long-term benzodiazepine usage on their own by making them aware of the adverse effects. This type of intervention is successful in about one fifth of patients. Gradual discontinuation programs are more extensive interventions in order to help those patients who are unable to discontinue benzodiazepine use on their own. These programs are successful in two-thirds of the patients and can be combined with additional pharmacological or psychological treatment. Once the withdrawal symptoms have diminished, most patients have an improved psychological functioning compared to when they were using benzodiazepines. Furthermore, no increase in medical consumption has been found. PMID- 11484432 TI - [Diagnostic image (46). Post-radiation angiosarcoma of the breast]. AB - A 71-year-old woman developed an angiosarcoma of the breast, five years after an adenocarcinoma had been removed and additional local radiation therapy with 70 Gy had been given. PMID- 11484433 TI - [From gene to disease; 'maturity-onset diabetes of the young' (MODY), monogenetic inheritable forms of diabetes mellitus]. AB - Maturity-onset diabetes of the young (MODY) exhibits an autosomal dominant pattern of inheritance and can be divided in at least five subtypes (MODY 1 to 5), each subtype being caused by mutations in a specific gene. The unambiguous molecular diagnosis of the specific MODY subtype facilitates an early diagnosis of diabetes and can help to reduce the development of diabetic complications. Furthermore, MODY2 patients generally have a milder clinical course and fewer complications than MODY3 patients, who consequently require a more aggressive therapeutic approach. PMID- 11484434 TI - [No effect of aircraft crash near South Limburg (the Netherlands) on self perceived health, but an increase in perceived environmental nuisance by the inhabitants]. AB - OBJECTIVE: To investigate the influence of an aircraft crash near the South of Limburg, the Netherlands, on the perceived physical and psychological health and annoyance of local residents. DESIGN: Controlled, blinded, follow-up questionnaire survey. METHOD: Three months before an aircraft crash occurred, randomly selected local residents and control subjects who lived approximately 20 km from the crash site were asked to complete a health questionnaire. The study was repeated one year after the crash. The aim of the survey was not mentioned in the questionnaire. The questionnaire asked about demographic features, physical health, psychological health and environmental nuisance. In the first survey, 544/926 (59%) questionnaires were successfully returned, and after the disaster this figure was 677/900 (75%). Linear regression modelling was used to check whether or not any differences in outcome were due to confounding demographic variables. RESULTS: There was no significant difference in the perceived physical and psychological health of local residents before and after the aircraft crash. A year after the crash, the nuisance level of aircraft noise reported by local residents had markedly increased. Aircraft noise was the major cause of disturbance. More than 1 in 3 residents perceived daily aircraft noise as a nuisance compared with 1 in 6 before the crash. The scores of local inhabitants for annoyance due to aircraft odour and dust doubled after the crash. Compared with control subjects, local inhabitants scored more than 20 times higher for annoyance due to aircraft odour and 40 times higher for annoyance due to dust. The reported nuisance caused by road traffic noise, odour and dust also increased, although to a lesser extent. CONCLUSION: Health was not influenced by the aircraft crash. A year after the crash, there was a strong drop in tolerance of aircraft nuisance. There was also a reduction in the tolerance of comparable nuisance due, for example, to road traffic, although this change was less marked. PMID- 11484436 TI - [Diagnostics and treatment of distal bile duct or pancreatic carcinoma; guidelines of the Amsterdam and Twente Comprehensive Cancer Centers]. AB - A project on the diagnostic work-up and treatment of distal bile duct and pancreatic carcinoma was carried out in the region covered by the Comprehensive Cancer Centre in Amsterdam, the Netherlands. It consisted of (a) describing the diagnostic work-up in the area, (b) developing the guidelines, (c) spreading the recommendations. The implementation and application of the guidelines are currently being evaluated. PMID- 11484435 TI - [Diagnosis and treatment of pancreatic carcinoma in the region of Amsterdam Comprehensive Cancer Care Center in 1997]. AB - OBJECTIVE: To describe the diagnostic work-up and treatment of patients with a pancreatic carcinoma in the Amsterdam area, the Netherlands, particularly in general hospitals. DESIGN: Retrospective, descriptive. METHOD: During 1997, 286 patients with a pancreatic carcinoma were diagnosed in 20 hospitals in the Amsterdam area. Diagnostic work-up and treatment data were collected from the medical records and analysed. RESULTS: Ninety percent of the patients presented in one of the 17 general hospitals (n = 252; 132 men and 154 women; mean age: 70 years). Thirty-five percent of them underwent diagnostic investigations which did not focus directly on pancreatic pathology. Ultrasound was performed in 97% of patients (4% in combination with Doppler) and CT in 60% (4% spiral CT). Endoscopic retrograde cholangiopancreatography (ERCP) was performed in 39% of these patients and endoprostheses were only inserted in half the cases. Thirty five percent of the patients who underwent both CT and ERCP underwent ERCP first. Ninety-nine patients (39%) were referred to a reference hospital for further investigation or treatment. The period between the first investigation and the histological diagnosis was 4 weeks. CONCLUSION: In the diagnostic work-up of patients with a pancreatic carcinoma, invasive diagnostic procedures were often performed before the non-invasive tests. Spiral CT was used minimally and ERCP was frequently performed without subsequent biliary drainage. The mean duration of diagnostic work-up was relatively long. PMID- 11484437 TI - [Initial experience with selection procedures for admission to medical school]. AB - Admittance to a medical school in the Netherlands has for decades been based on a grade point average weighted lottery system of secondary school leavers. Since 2000, the Dutch Higher Education and Scientific Research Act has given medical schools the option of selecting candidates. In 2000, two of the eight Dutch medical schools started selection experiments for 10 percent of their places. Leiden University Medical Center invited school leavers who had studied a more varied range of extra subjects to attend a 10-day summer school. All 54 candidates were ranked on the basis of assessments and tests; 24 of them were admitted. Utrecht University invited students with a higher education degree to a selection day. An application form, a structured interview and a questionnaire determined the ranking of 53 candidates; 24 of them were admitted. Both schools were satisfied with the manner in which the selection procedure worked. However, it is not yet possible to draw any definite conclusions about the effectiveness of the selection procedure. PMID- 11484438 TI - [Responsibility circumvented with a set time for medical record keeping]. PMID- 11484439 TI - Subjective outcomes after knee arthroplasty. PMID- 11484440 TI - [Ultrasound safety in ultrasound diagnosis--what should users know?]. PMID- 11484441 TI - [Ultrasound diagnosis of bilharziasis]. AB - AIM: Schistosomiasis (Bilharzia, Bilharziasis) is one of the most prevalent tropical diseases, with an increasing number of cases being imported into Europe. Sonography is among the most valuable diagnostic tools for schistosomiasis related organ lesions. This review outlines typical findings and their pathophysiological context. METHOD AND RESULTS: Bilharziasis of the urinary tract, usually due to Schistosoma (S.) haematobium, leads to diffuse or localized wall thickening of the bladder and the distal ureter with typical sonographic features. Upper urinary tract obstruction and--rarely--bladder carcinoma may complicate the course and can also be detected sonographically. The other species (S. mansoni, S. japonicum, S. mekongi, S. intercalatum) primarily cause (entero-) colitis; the value of sonography in this condition is yet undefined. In later stages, fibrotic liver involvement with portal hypertension may develop (hepatosplenic schistosomiasis), leading to typical ultrasound features which are nearly pathognomonic under endemic conditions: severe periportal echogenicity with S. mansoni, and a peculiar "network pattern" of echogenic septa with S. japonicum. Sonographic indicators of portal hypertension may be identified and graded. CONCLUSION: In endemic areas in the tropics, sonography with simple portable machines offers a unique opportunity to investigate morbidity on the community level non-invasively in large field surveys; it has thus become an important tool of clinical and epidemiological research. PMID- 11484442 TI - [Color duplex ultrasound imaging of renal arteries and detection of hemodynamically relevant renal artery stenoses]. AB - AIM: Although colour coded Doppler ultrasound (CCDU) is established as a non invasive diagnostic tool for detecting renal artery stenoses (RAS), no uniform criterion for defining a hemodynamically relevant stenosis (i.e. angiographic stenosis of > or = 70% of the diameter) exists. We therefore investigated the predictive value of the renal-aortic flow-velocity ratio (RAR) > 3.5 m/s and a difference of < 0.05 between the left and right side for the resistance index according to Pourcelot (dRI) in the detection of a relevant RAS. PATIENTS AND METHODS: We analysed 500 consecutive CCDU examinations of patients with hypertension retrospectively. An RAR > 3.5 and/or a lateral inequality of the RI < 0.05 were used as stenosis criterion. RESULTS: In 448 patients (90%) both renal arteries could be found, in 11 patients (2%) only the right artery, in 6 patients (1%) the left artery, and in 35 patients (7%) no renal artery was detectable. In 98 patients (19.6%), RAS was diagnosed, 69 (71%) of them underwent angiography. 38 patients presented an RAR > 3.5 plus dRI > 0.05. In 29 of these, angiography was performed. 96% of them presented with an RAS of > or = 70% and 4% showed an RAS of 40-69% (specificity 97%, sensitivity 76%). In 54 patients the RAR was > 3.5, but dRI < 0.05. 24% of the patients undergoing angiography (n = 37) presented with an RAS of > or = 70%, 68% with an RAS of 40-69%, and 8% with an RAS of < 40% (specificity 60%, sensitivity 100%). 44 hypertensive patients who underwent angiography after a CCDU examination not suggesting the presence of RAS were used as control group. CONCLUSION: An experienced physician using a high quality colour-coded duplex-machine can reliably detect the renal arteries. The presence of RAS can be diagnosed with certainty by CCDU applying the criterion of RAR > 3.5, but the diagnosis of a one-sided haemodynamically relevant RAS can only be certain if the criterion of dRI > 0.05 is used in addition. PMID- 11484443 TI - Impaired compliance of the intracranial vessels in complicated childhood migraine. Demonstration by transcranial Doppler-sonography--a vascular model approach. AB - AIM: To increase the diagnostic value of Doppler sonography a uniform lumped parameter model of pulsatile blood flow in human arteries was derived from the serial and/or parallel connection of parallel oscillatory circuits. METHODS AND PATIENTS: In this setting the current (I) in the ohmic resistor (R) represents the volume flow in the vessel, the voltage (V) the blood pressure. The vessel compliance corresponds to the capacitance (C), the inductivity (L) represents the accelerated mass. The flow velocity (v) which is assessable non-invasively is given by v = I/A, where A is the cross-sectional area of the vessel. Mathematically, the model represents a two-dimensional Fourier-series of the input signal by the segments. Thus it seems to be evident that almost all forms of flow and pressure pulses can be generated. The equations of the model were solved numerically. Doppler-sonographically measured flow velocities in the ascending aorta were used as input signal. The parameters for A, R, L and C were optimized to gain the measured flow spectrum in the target vessel as model response. RESULTS: Application in 14 children (aged 3-16 yrs.) with complicated migraine during a symptom-free interval revealed a significantly lowered cross sectional area of the middle cerebral artery on the side of the higher flow velocities. For both middle and posterior cerebral arteries, a decreased vessel compliance (C) (mean -30%) was found, whereas values for R and L were within normal limits. CONCLUSION: These findings suggest an altered vascular compliance even between attacks. This may contribute to the pathogenesis of migraine through increased cerebral impedance resulting in impaired cerebral perfusion. PMID- 11484444 TI - [Ultrasound characteristics of lateral cervical space-occupying lesions]. AB - Sonography of the head and neck mainly focuses on tumours of the lateral neck. Even though most of these masses are enlarged lymph nodes, other tumors have to be considered. AIM: To determine whether different pathological cervical masses display a characteristic sonographic morphology. METHOD: On the basis of 530 sonographical findings in the neck, typical ultrasound characteristics either in B-scan or in colour coded sonography were demonstrated. RESULTS: Apart from enlarged lymph nodes, it was possible to differentiate other occurrences such as branchial cysts, abscesses, laryngoceles, lipomas, neurinomas, glomus tumours, paragangliomas and also fibromas by their specific ultrasound appearance. CONCLUSION: Knowledge of characteristic ultrasound morphology allows the early detection of tumours and facilitates the distinction between benign and malignant lesions as well as malformations. Further diagnostic procedures and therapeutical strategies can therefore be planned more adequately. PMID- 11484446 TI - [Value of ultrasound in microlithiasis of the testis in andrological patients]. AB - PURPOSE: The objective of this retrospective study is to show the importance of sonography in andrological patients with testicular microlithiasis (TM). PATIENTS AND METHODS: 1314 male patients were seen to our andrological clinic in the course of one year. The age range of these patients was 25 to 39 years (mean age 32 years). All patients underwent testicular sonography as well as a sperm analysis. RESULTS: 284 patients showed normozoospermia without any evidence of TM. Of the remaining 1030 patients with a pathological spermiogram, 8 were shown to display more than 10 echogenic foci per transducer field in both tests. 1 patient suffering from an already palpable testicular tumor only showed, unilateral, unifocal calcification. Another patient who had suffered from a maldescensus testicle in his early childhood discharged only one unifocal calcification. Tumor markers including AFP and beta-HCG were normal in 9 patients, but elevated in 1 patient suffering from a testicular tumor (AFP: 73 kU/l; beta-HCG: 10.6 U/l). The hormonal status was normal in 6 patients and pathological 4 patients with the diagnosis of OAT-syndrome. CONCLUSION: TM is a rare condition even in andrological patients. Nevertheless, a thorough scrotal sonography is mandatory in order to rule out testicular malignancy. PMID- 11484445 TI - [Ultrasound endometrium follow-up during tamoxifen treatment: Really not reliable or useful after all?]. AB - AIM: To investigate whether an examination of the endometrium of women treated with tamoxifen (TAM) is useful or not. METHOD: 40 breast cancer patients who displayed a thickened endometrium of > 8 mm and/or vaginal bleeding were included in the study. They received daily TAM adjuvantly. Histologic clarification by hysteroscopy and D&C was recommended for patients with an endometrium of > 8 mm or vaginal bleeding. RESULTS: In our collective, the mean endometrial thickness was 13.7 +/- 5.6 mm (SD). 32 patients underwent a histological examination. Most had a benign lesion; in 2 cases we merely found a cystic atrophy (11 mm, 18 mm), 2 displayed atypical tissue (13 mm, 25 mm) and 2 an endometrial cancer (19 mm, 33 mm). All patients with atypical tissue or cancer had an endometrial thickness markedly above the norm, but 3 of them were not bleeding. No linear correlation between thickness of the endometrium and duration of TAM intake was found. CONCLUSION: To detect early premalignant or malignant changes of the endometrium, we recommend histological examination by hysteroscopy and dilatation and curettage when the endometrium is > 8 mm thick, even in the absence of symptoms. Therefore, these patients should have regular examinations by transvaginal ultrasound once or twice a year. Moreover, continuing regular screening of the endometrium for years after termination of tamoxifen-therapy is also to be recommended. PMID- 11484447 TI - [Thermal effects of ultrasound]. AB - AIM: Diagnostic ultrasound is a powerful, versatile and safe investigation tool in modern medicine. Methods are described, and conclusions are drawn and recommendations given for a responsible use of diagnostic ultrasound. METHOD AND RESULTS: Because of the increasing ultrasonic power penetrating the body, diagnostic techniques cannot always be applied without risk. The increase in temperature induced by ultrasound absorption should be considered a potential hazard in certain applications. The paper presents the physical basis for a quantitative estimation of the risk of a specific treatment plan. Mathematical models and the thermal index are described. CONCLUSION: Our findings and recommendations form a viable basis for the responsible use of diagnostic ultrasound. PMID- 11484448 TI - [Intercostal percutaneous ultrasound guided decompressive gastrostomy (PUG)--a case report]. AB - Percutaneous gastrostomies (PG) are nowadays usually placed endoscopically (PEG). Due to stenoses in the pharynx or the esophagus or in case of a lack of diaphany, other methods for the placement of PG such as computed tomography (CT-) guided or ultrasound-guided PG (PUG) are performed. We present a case of a 63-year old male patient with a retroperitoneal liposarcoma and multivisceral resection who presented with the symptoms and signs of an ileus due to metastatic occlusion of the small bowel. The placement of a decompression-PEG was impossible due to interposition of colon between the stomach and the abdominal wall. After filling the stomach with water via a naso-gastral decompression tube, the stomach could be visualized at the left thoracic wall, since the spleen and the left kidney had been removed at prior surgery. A transcostal decompression-PUG was inserted. The intervention described is the first published ultrasound guided transcostal decompression gastrostomy and is an example for the growing significance of interventional ultrasound. PMID- 11484449 TI - [Reception of parents of hospitalized children]. PMID- 11484450 TI - [Posterior fossa hemorrhage in newborns following vaccum extractor delivery]. AB - AIM: To emphasize the risk of posterior fossa hemorrhage in newborns following vacuum extraction. PATIENTS AND METHODS: Over a period of 26 months (September 1996-December 1998), seven patients who underwent delivery with the vacuum extractor had symptoms of brain stem compression, related to posterior fossa hemorrhage. They were referred to the neonatal intensive care unit. Some parameters had been analyzed: gestational period, delivery circumstances, gestational age, parameters of newborn, indications of vacuum extraction and other paraclinical investigations (biological and radiological). RESULTS: Ultrasound scan revealed intracranial hemorrhage in five cases of seven; mean time of diagnosis was 10 hours of age. All patients presented symptoms of brain stem compression. Mean period of follow-up was 22 months: six of seven patients had a normal neurodevelopment. One patient had a cerebellar ataxia, another one a palsy of the IIIrd cranial nerve. CONCLUSION: Analyses of posterior fossa by ultrasound scan should be made for newborns delivered by vacuum extractor, especially if they had symptoms of brain stem compression. PMID- 11484451 TI - [Treatment of acute group A beta-hemolytic streptococcal tonsillitis in children with a 5-day course of josamycin]. AB - MATERIAL AND METHODS: In this randomized open study, 325 children aged two to 15 years with acute tonsillitis and a positive test of GA beta H streptococcal antigen were treated with josamycin 50 mg.kg-1.day-1 b.i.d for 5 days, or penicillin 50,000 to 100,000 IU/day t.i.d for 10 days. Clinical assessments and throat cultures for GA beta HS isolation were performed at the inclusion visit (V1), at the end of treatment visit (V2: day 12 for all patients) and at the follow-up visit (V3: day 30). In case of positive GA beta HS culture, the bacterial DNA by RFLP was performed to differentiate between the persistence (presence of original strain at V2), relapse (eradication at V2 and acquisition of same strain at V3) and reinfection (eradication at V2 and acquisition of different strain at V3). RESULTS: Two hundred and twenty-three patients were included in the bacteriological and clinical criteria per protocol analysis. At V2, eradication rates were comparable: 82% in josamycin and 80% in penicillin patients; clinical cure rates were 90% and 89%. At V3, relapse of GAS assessed only on clinically and bacteriologically cured patients at V2 occurred in 12% of josamycin patients and 12.8% of penicillin patients. Tolerance was good; 14% and 10% of josamycin and penicillin patients respectively experienced an adverse event. CONCLUSION: In this non-inferiority study, the efficacy of a 5-day course of josamycin is comparable to reference treatment in GA beta HS tonsillitis in children. PMID- 11484452 TI - [Echocardiographic aspects in pediatric patients with sickle cell disease]. AB - BACKGROUND: Cardiovascular involvement is not well studied in children with sickle cell disease. The aim of this study was to evaluate the echocardiographic parameters of children with sickle cell disease. PATIENTS AND METHODS: We performed a transversal and case-control study including 80 subjects of six months to 16 years of age divided into four groups of 20 children each: heterozygous sickle cell disease, homozygous sickle cell anemia, anemia of other causes than sickle cell, and healthy children. All children had a complete physical examination, biological screening including hemogram and hemoglobin electrophoresis, chest x-ray, electrocardiogram, and Doppler echocardiogram. Data were compared using the chi 2 method and the Student's t-test. RESULTS: The mean age was 8.5 years and the sex-ratio was 1. Echocardiographic abnormalities were observed in seven patients with anemia unrelated to sickle cell, 15 children with heterozygous anemia and all the homozygous patients. The main abnormalities were: left ventricular enlargement (ten homozygous patients, one heterozygous subject, five of the patients with another cause of anemia), increased contractility of the heart (18 homozygous patients, seven heterozygous patients, five in the anemia group) and mild to moderate mitral or tricuspid regurgitation (12 homozygous, five heterozygous and five patients in the anemia group). One homozygous child had a dilated and hypokinetic cardiomyopathy with pulmonary hypertension. The parameters of left ventricular systolic function and left heart chamber dimensions were lower in the control group (P < 0.04). DISCUSSION: This study shows the frequency of heart chamber dilatation with left ventricular hyperkinesis usually described in the literature. The lack of case of chronic cor pulmonale may be due to the young age of our patients. One case of dilated and hypokinetic cardiomyopathy suggests that other causes than anemia should be considered, particularly myocardial ischemia, which could not be demonstrated because of our limited investigative methods. CONCLUSION: These results emphasize the frequency of the heart involvement in sickle cell disease, particularly in the homozygous type, and point out the importance of the cardiologic screening of these patients. PMID- 11484454 TI - [Post-traumatic hepatic artery pseudoaneurysm. A case report]. AB - CASE REPORT: We report a case of traumatic hepatic artery pseudoaneurysm in a 9 year-old child. The diagnosis was made by duplex sonography and helical CT angiography. Arteriography confirmed and managed the lesion with transcatheter embolization. CONCLUSION: The traumatic hepatic artery pseudoaneurysm is an uncommon complication of liver trauma in children whose diagnosis is made by duplex sonography and CT angiography. Endovascular embolotherapy is the treatment of choice. PMID- 11484453 TI - [Cardiovascular complications of Kawasaki syndrome: results of a French multicenter study]. AB - PATIENTS AND METHODS: Between March 1, 1995 and February 29, 1996, a multicentric prospective study was conducted in France in order to analyze the cardiovascular complications in Kawasaki syndrome, and to describe the echocardiographic features and the outcome of coronary lesions. Forty-nine cases of Kawasaki syndrome were observed. RESULTS: There were 32 boys and 17 girls (sex ratio: 1:9). The age at diagnosis was under one year in ten cases (20.4%), between one and five years in 27 cases (55.1%) and more than five years in 12 cases (24.5%). The complete diagnostic criteria were present in 42 cases (85.1%). Forty-five children (91.8%) were given intravenous immunoglobulin treatment but only 20 (40.8%) received this treatment within the seven days following the onset of the illness. Cardiovascular complications consisted of: pericardial effusion in 12 cases (24.5%), coronary dilation in seven cases (14.3%), coronary aneurysms of moderate size in seven cases (14.3%,) with hypokinetic left ventricle in two cases. No death was reported. All patients with coronary dilation and four patients with coronary aneurysms had a normal size of coronary arteries at echocardiography within the nine months of the follow-up. Among the three other patients, after a follow-up of two years, one still has a small coronary aneurysm and two have a normal size of coronary arteries. CONCLUSION: Despite a delayed administration of immunoglobulin therapy in the majority of patients in this study, outcome of coronary lesions was favorable and severe cardiac complications were rare in the acute phase of the Kawasaki syndrome. PMID- 11484455 TI - [Takayasu's disease: interest in methotrexate treatment]. AB - Takayasu's disease is a nonspecific aortic arteritis that affects mostly young women and sometimes children. Usual treatment consists of early and prolonged steroids. Some patients do not respond to this treatment, become steroid dependent or suffer from side effects. We report a case in which methotrexate proved to be effective. CASE REPORT: A 6-year-old girl presented with Takayasu's disease with elevated blood pressure of renovascular origin. Corticosteroids controlled the inflammatory syndrome but not the renal involvement and stopped the growth. Methotrexate (10 mg/m2/week) resulted in the control of the disease, the reduction of steroids and normal growth. CONCLUSION: In Takayasu's disease, steroids should be given as first-line therapy. In case of failure, side effects or steroid dependency, small doses of methotrexate may facilitate the disease's control and weaning from the steroids. PMID- 11484457 TI - [Hypernatremic dehydration and breastfeeding]. AB - Neonatal hypernatremic dehydration due to breast feeding was rarely reported in the French-language literature. CASE REPORT: The authors report hypernatremic dehydration in a 10-day-old exclusively breast-fed infant. The course was favorable. Insufficient breast milk production has been reported for 30 years. CONCLUSION: Early discharge from maternity units is frequent nowadays. Breast-fed newborns without a positive weight gain at discharge should be checked for their weight within the first ten days of life. PMID- 11484456 TI - [Bilateral vas deferens agenesis and inguinal hernia in a child. A rare, early presentation of cystic fibrosis]. AB - Epididymal and ductal anomalies can be discovered incidentally during inguinal herniorraphy in children. The congenital bilateral absence of vas deferens is frequently associated with cystic fibrosis. CASE REPORT: This agenesia of vas deferens was detected in a 5-month-old boy who underwent an inguinal herniorraphy. Although the child did not present any symptoms, he actually presented cystic fibrosis: the sudoral test showed high levels of chloride (95 mmol/L) and an isolated homozygous delta F 508 deletion on the gene CFTR was evidenced on genetic investigations. CONCLUSION: The congenital bilateral absence of vas deferens is the most frequent anomaly of the male genital tract discovered in adults investigated for azoospermia. Relations with cystic fibrosis are well established but congenital bilateral absence of vas deferens discovered during infancy is an exceptional situation that requires genetic investigations to show evidence of a likely underlying cystic fibrosis. PMID- 11484459 TI - [Quality of life in children: what is a good evaluation tool?]. AB - The tools most often used to measure subjective data in health, especially quality of life, are based on psychometry. The object of this article is to provide as simply as possible an explanation to the specific methodological aspects of this field. As an example, the child version of the Children Health Questionnaire (CHQ-CF87) is used to study how to demonstrate the qualities required for a good assessment tool: acceptability in practice, responsiveness, reliability and validity. These qualities must be reassessed when a given tool is translated into another language. PMID- 11484458 TI - [Immunization for children treated for solid tumors: what are the guidelines?]. AB - There is no agreement on immunization of children treated with chemotherapy (CT) for solid tumors. Based on a review of the literature, we have attempted to establish guidelines on this subject. Except for hepatitis B vaccine, there is no argument to support the use of vaccine during CT. After a standard CT, a 3-month washout period appears to be necessary before starting an immunization program for a child not previously vaccinated, or to proceed with the recommended booster injections for diphteria anatoxin, tetanus vaccine, poliomyelitis inactivated vaccine, pertussis vaccine, and haemophilus influenza type b vaccine if the child is less than 5 years old. For mumps, measles, and rubella live vaccines, a longer post-CT washout of 6 months is suggested for the initial immunization, or for a revaccination of a child proved to be negative for all three serologies. Following high-dose CT a minimal 12-months term and a normalization of the blood lymphocytes count is necessary before planning booster injections once having checked for antidiphteria, tetanic, polio, measles, mumps, rubella and +/- haemophilus antibody titles. We don't find any reason to recommend a systematic varicella immunization in pediatric oncology. Pneumococcal vaccine is recommended in case of asplenia. Any other vaccination (BCG, influenza, yellow fever) must be evaluated individually. PMID- 11484460 TI - [Children also die]. AB - Palliative care has recently been developed in adults. However, children die as well and quite a few articles have been published on this special issue. The object of this article is to summarize the available literature in order to call for the development of clinical policies and minimum standards adapted to French pediatrics. PMID- 11484461 TI - [Dental anomalies following anticancer chemotherapy]. AB - Chemotherapy administered during childhood may induce dental abnormalities, such as acquired amelogenesis imperfecta, microdontia, hypodontia and altered root morphology. The magnitude of the defect varies according to the cytotoxic agents, the duration of their use and the stage of tooth development at the time of chemotherapy. Patients who received high-dose chemotherapy before the age of 5 are particularly concerned. The dental supervision of these children is based upon three orthopantomograms: the first one has to be performed before starting chemotherapy and will be used as a reference; the second is done soon after the drug therapy in order to evaluate the first consequences; the third is performed after the eruption of all permanent teeth (age 12-13 in average) in order to determine the dental abnormalities. In case of hypodontia, orthodontic treatment must be considered, but it is necessary to take into account the fact that it may increase the risk of root resorption. Preventive dental care is important for these children. It involves meticulous oral hygiene and frequent dental visits to assess and maintain dental health. PMID- 11484462 TI - [Tracheomalacia and tracheal dyskinesias]. AB - Tracheomalacia is characterized by an expiratory tracheal collapse. The revealing clinical features usually occur after a symptom-free period during the first year of life (during the first three months in 60% of cases): mainly stridor, wheezing, chronic cough, apnoeic attacks, and difficult breathing. Tracheoscopy allows the differentiation of primary tracheomalacia and tracheomalacia secondary to pressure on the trachea from the vascular ring or mediastinal tumour. Tracheomalacia is also frequently associated with oesophageal atresia. Surgical treatment is indicated in severe and secondary forms. PMID- 11484463 TI - [A plea for modifying the term acute infantile hemorrhagic edema]. PMID- 11484464 TI - [Diagnosis of acute fetal distress and cesarean delivery in unfavorable conditions]. PMID- 11484465 TI - Caries prevalence in Belgian children: a review. AB - A review of epidemiological surveys on dental caries prevalence, published between 1980 and 1999 in Belgian children, was compiled through a literature search. The number of studies performed in Belgium to date is limited. Methodological differences and confounding factors, especially socio-demographic influences, limit national comparisons of caries prevalence data. Although exact comparisons are difficult, data suggests a decline in caries prevalence in 5, 7 and 12 year-old Belgian schoolchildren in the last 20 years. In the primary dentition dmft values have decreased from 2.66 (1981) to 1.38 (1994) in 5-year olds and from 4.1 (1983) to 2.24 (1996) in 7-year-olds. In 12-year-olds DMFT values in the permanent dentition have decreased from 3.9 (1983) to 1.93 (1994). WHO goals for the year 2000 appear to have been already reached in Flanders, with a recent estimate of 1.93 for DMFT in 12-year-olds and 56% of children being recorded as caries free at the age of 5. Continuing efforts are needed to screen the oral health of different age groups but standardised criteria and sampling procedures should be used if benefits are to be gained from national and international comparison. Data has often been limited to small selected areas and information representing the entire community of Flanders or Wallonia would be of particular value. PMID- 11484466 TI - Root resorption in retained deciduous canine and molar teeth without permanent successors in patients with severe hypodontia. AB - AIMS: The ability to predict the morbidity of retained deciduous teeth with no permanent successors, a characteristic of hypodontia, would be of considerable value in treatment planning, but is hampered by lack of data. METHODS: This problem was studied using 356 orthopantomogram radiographs (OPGs) from the records of 249 patients who had attended a specialist hypodontia clinic, and had retained deciduous teeth with no permanent successors. Due to their clinical importance, canine and molar teeth were chosen for examination. Resorption was assessed subjectively by three experienced clinicians. RESULTS: Un-weighted Kappa values for reproducibility were > 0.8, and for interobserver error 0.60-0.83. Gender related differences were minimal. Regardless of gender or radiographic age, the lower canines appear to show the least amount of resorption and the upper first molars the most. The upper and lower second molars have particularly unpredictable life spans. Whilst the lower first molars have a predictable life span that is poor, the life span for the upper first molars is slightly worse. CONCLUSIONS: Lower canines have a predictable life span that appears to be good, as do upper canines, but of lesser duration than lower canines. Molars have poorer and less predictable life spans. PMID- 11484467 TI - Celiac disease and mineralisation disturbances of permanent teeth. AB - In a study from Finland in 1986 it was shown that celiac disease was often associated with tooth enamel defects of permanent teeth. This study also showed a strong association between the time of gluten challenge in the diagnostic procedure and enamel defects. In the current study, dental examinations were carried out for a group of 40 children and adolescents suffering from celiac disease diagnosed according to the criteria of The European Society of Paediatric Gastroenterology and Nutrition (ESPGAN) at the Department of Paediatrics, Hospital of Uddevalla, Sweden. A control group made of 40 healthy children of the same age, sex and living area was examined in the same way. The results failed to show disturbances of the same type, degree of severity or frequency as was reported in Finland and no statistically significant differences concerning enamel defects were found between the patients with celiac disease and the controls. PMID- 11484468 TI - Vegetarian children and dental erosion. AB - BACKGROUND: There have been recent changes in teenage lifestyle and diet. The increasing consumption of soft drinks and foods containing significant acidic components may play a role in the development of dental erosion. OBJECTIVE: The aims of this investigation were firstly to assess the prevalence of vegetarian children in a cluster random sample of 14-year-old children in Birmingham, United Kingdom. Secondly, to determine the prevalence of dental erosion in these children, and thirdly, to see if there were any differences between vegetarian and non-vegetarian children in the prevalence of dental erosion and dietary intake. DESIGN: A cluster random sample of 418 14-year-old children (209 males and 209 females) were examined from 12 different schools in Birmingham, United Kingdom; a dietary questionnaire was completed and the levels of tooth wear were recorded using a modification of the (TWI) index. All data were analysed using SPSS with t-test and Chi-square analysis. Significance was accepted at the P < 0.05 level. RESULTS: The results showed that 10% of the children were vegetarian; 52% of them had low dental erosion and 48% moderate dental erosion. Statistically there were no significant differences between vegetarian and non-vegetarian children in the prevalence of erosion; however, there were significant differences in some food and drink consumption. CONCLUSIONS: It was concluded that dental erosion is common in teenage children, but there were no significant differences in prevalence between vegetarian and non-vegetarian children. PMID- 11484469 TI - A quantitative study of the effect of pumicing and etching on the remineralisation of enamel opacities. AB - AIMS: The aim of this study was to investigate the effects of pumicing and acid etching on the remineralisation of enamel opacities using quantitative measurement methods. METHODS: Teeth with diffuse opacities were subjected to a treatment sequence of: (a) pumicing for 30-35 seconds (b) etch with 37.5% phosphoric acid for 2 minutes, (c) pumicing for 30-35 seconds and (d) etch for 1 minute, or served as controls. Both groups were then placed in a remineralising solution containing 1.5 mmol/L Ca2+ for 3 weeks. Mineral content changes were measured using contact microradiography and x-ray microtomography. In addition, the reflectivity of the opacities was monitored with infrared reflectance. RESULT: The treatment sequence removed 34 +/- 4 microns of the surface enamel but no mineral loss was observed in the subsurface layer. The treatment sequence enhanced the formation of a new remineralised surface layer with a mean thickness of 22 +/- 3 microns. This new surface layer was less reflective. CONCLUSION: Pumicing and acid etching can be used as a simple technique for the conservative treatment of enamel opacities. PMID- 11484470 TI - Correction of anterior dental crossbite with composite as an inclined plane. AB - OBJECTIVE: The purpose of this study was to evaluate a passive method to correct anterior crossbite of only one incisor by constructing a composite inclined plane. SAMPLE AND METHOD: A total of 35 children aged 7-11 with a crossbite of only one incisor tooth were selected for this study. An inclined composite block 3-4 mm in size and 45 degrees to the longitudinal axis of the tooth, was constructed on the incisal edges of the related mandibular incisors. The occlusion was checked so that the only contact between both arches was at the level of these incisors. RESULTS: At the end of one week, 33 cases of crossbite were corrected by this method. Of the two cases which were not corrected one had a deep overbite and the second had a rotated incisor together with crossbite. CONCLUSION: The present results show that, a composite inclined plane is a useful technique to correct anterior crossbite of only one incisor tooth. PMID- 11484471 TI - Oral mucosal lichen planus in children. AB - Although lichen planus is a relatively common mucocutaneous disorder in adults, it has only rarely been described in children. Moreover, even less data has been published regarding mucosal lesions in children. Six case reports of childhood oral lichen planus are presented and the available literature reviewed. It is believed that this paper documents the largest series of cases of childhood oral mucosal lichen planus to be reported in the literature to date. Lichen planus should be considered in the differential diagnosis of oral mucosal white patches in children, particularly those of Asian origin. PMID- 11484472 TI - Facial features of widespread neuroblastoma: a case report. AB - A case report of stage IV neuroblastoma which presented with periorbital swelling and ecchymosis originally misdiagnosed as facial trauma. The child soon developed a sinister pancytopenia, which following extensive investigations was revealed to be due to an underlying neuroblastoma. Periorbital ecchymosis associated with neuroblastoma is termed 'raccoon eyes' and is a diagnostic trap for the unwary. PMID- 11484473 TI - Intranasal tooth as a complication of cleft lip and alveolus in a four year old child: case report and literature review. AB - Ectopic position of teeth is not rare. The presence of teeth have been reported in ovaries, testes, anterior mediastinum, and pre-sacral regions. In the maxillofacial region, teeth have been found in maxillary sinus, mandibular condyle, coronoid process, chin, nose, and even orbit. Approximately 50 cases of a tooth in the nasal cavity have been reported in literature. However, an intranasal tooth in cases of cleft lip and palate is comparatively rare. Intranasal teeth can cause problems such as nasal obstruction, chronic rhinorrhea and speech problems. Sometimes however, they are totally symptom-free. We present here an interesting case of an intranasal tooth in a four year-old-boy, who was operated on for cleft lip and alveolus at 6 months of age. The intranasal tooth did not cause any symptoms. The tooth was extracted under general anaesthesia when it was found to be very loosely attached to the nasal mucosa. The case is discussed in the light of relevant literature on intranasal teeth in cases of cleft lip and palate. PMID- 11484474 TI - Orofacial findings in the Klippel-Trenaunay syndrome. AB - The Klippel-Trenaunay syndrome is a triad of congenital anomalies characterised by haemangiomas, varicosities, and unilateral bony and soft tissue hypertrophy. Hypertrophy usually affects one distal limb, but trunk or face may be affected. Cutaneous haemangiomas (nevus flammeus) of varying extent and irregular contour are often present in the hypertrophic regions. Varicosities may also be part of the vascular lesions of the syndrome. Orofacial manifestations include facial asymmetry, jaw enlargement, and malocclusions as well as premature tooth eruption. Two cases of the Klippel-Trenaunay syndrome are presented here. Both of these show the typical hemifacial hypertrophy and premature eruption of teeth on the affected side. In the first case only the left mandibular region was affected. In contrast, in the second there was hypertrophy of the whole left side of the body including upper and lower jaws. This boy also suffers from congenital ideokinetic retardation, while the first was otherwise normal. Both cases differ from previously reported cases of the Klippel-Trenaunay syndrome in lacking any prominent facial nevus flammeus. In the first case there was also malformation of the crown of the first permanent molar on the affected side that has not been described previously. PMID- 11484475 TI - British Society of Paediatric Dentistry: a policy document on the use of amalgam in paediatric dentistry. PMID- 11484476 TI - Purification of archaeal chaperonin from Sulfolobus shibatae. PMID- 11484477 TI - Prevention of rhodanese aggregation by the chaperonin GroEL. PMID- 11484478 TI - Refolding of bovine mitochondrial rhodanese by chaperonins GroEL and GroES. PMID- 11484479 TI - Assay of malate dehydrogenase. A substrate for the E. coli chaperonins GroEL and GroES. PMID- 11484480 TI - Assay of chaperonin-assisted refolding of citrate synthase. PMID- 11484481 TI - Purification of yeast mitochondrial Hsp60. PMID- 11484482 TI - Preparation of recombinant human Hsp10. PMID- 11484483 TI - Purification of Hsp60 from Thermus thermophilus. PMID- 11484484 TI - Purification of the cytosolic chaperonin TRiC from bovine testis. PMID- 11484485 TI - Monitoring actin folding. Purification protocols for labeled proteins and binding to DNase I-sepharose beads. PMID- 11484486 TI - Folding assays. Assessing the native conformation of proteins. PMID- 11484487 TI - Purification of prefoldin. PMID- 11484488 TI - Purification of GimC from Saccharomyces cerevisiae. PMID- 11484489 TI - Analysis of eukaryotic molecular chaperone complexes involved in actin folding. PMID- 11484490 TI - Purification of GroEL from an overproducing E. coli strain. PMID- 11484491 TI - Purification of GroES from an overproducing E. coli strain. PMID- 11484492 TI - Purification of the Gp31 Co-chaperonin of bacteriophage T4. PMID- 11484493 TI - Removing trace fluorescent contaminants from GroEL preparations. PMID- 11484494 TI - Assembly and disassembly of GroEL and GroES complexes. PMID- 11484495 TI - GroEL/GroES interaction assayed by protease protection. PMID- 11484496 TI - Determination of chaperonin activity in vivo. PMID- 11484497 TI - Interaction of nonnative polypeptide substrates with the Escherichia coli chaperonin GroEL. PMID- 11484498 TI - Histologic classification of penile carcinoma and its relation to outcome in 61 patients with primary resection. AB - A retrospective review of the clinical and pathologic features of 61 cases of penile squamous cell carcinoma (SCC), all treated by primary surgical resection at the Memorial Sloan Kettering Cancer Center during the period 1949-1992, was undertaken. Inguinal lymph node dissection material was evaluated in 40 cases. All carcinomas were of squamous cell type and were classified as follows: usual type, 36 cases (59%); papillary, not otherwise specified (NOS), 9 cases (15%), basaloid, 6 cases (10%); warty (condylomatous), 6 cases (10%); verrucous, 2 cases (3%), and sarcomatoid, 2 cases (3%). A high rate of nodal metastasis and poor survival were found for the basaloid and sarcomatoid neoplasms (5 of 7 patients with metastasis, 71%, and 5 of 8 dead of disease, 63%). Only 1 patient with a verruciform tumor (defined as a tumor of nonspecific papillary, warty, or verrucous type) had inguinal node metastasis and none died from penile cancer. An intermediate rate of metastasis and mortality (14 of 26, 54%, and 13 of 36, 36%, respectively) was found for typical SCC. Penile carcinomas are morphologically heterogeneous, and there is a correlation of histologic type and biologic behavior. This mandates accurate histologic subtyping by the pathologist. PMID- 11484499 TI - Biovularity and "coalescence of primary follicles" in ovaries with mature teratomas. AB - Several theories have been postulated regarding the origin of ovarian teratomas, including incomplete twinning, neoplastic proliferation of sequestered totipotent blastomeres or primordial cells, derepression of totipotent genetic information in the nuclei of somatic cells, and parthenogenetic development of germ cells. At present parthenogenetic development of ova is the most widely accepted theory, primarily because of the presence of a 46 XX karyotype in almost all mature teratomas. However, some authors have raised the possibility of fusion of ova in the mechanism of formation of ovarian teratomas. We report the results of a study on ovarian tissue adjacent to 31 teratomas to assess the frequency of biovularity, which could provide evidence favoring the last theory. On the whole we found biovularity in 26 ovaries of young patients (mean age, 27 years) with variable numbers of biovular follicles ranging from 1 in 4 cases to more than 10 in 2 cases; the number of biovular follicles depended on the quantity of ovarian tissue examined as well as on the total number of ova in the tissue. In multiple occasions 2 ova were included within a single follicle; in 24 ovaries the biovularity was correlated with coalescence of primary follicles characterized morphologically by an ovoid or hourglass-like shape that resulted from cohesion of 2 follicles. As control cases, 30 ovaries of patients with an average age of 28 years were examined (12 removed for endometriosis, 8 for serous cystadenoma, 7 for tubal pregnancy, and 3 for acute salpingo-oophoritis). Only 1 ovary with endometriosis contained a single biovular follicle. The results suggest that ovarian teratoma development may result from fusion of ova in ovaries containing biovularity and phenomena of coalescence of primary follicles. PMID- 11484500 TI - Lobular carcinoma in situ and infiltrating ductal carcinoma: frequent presence of DCIS as a precursor lesion. AB - Infiltrating ductal carcinoma (IDC) occurs frequently in patients with lobular carcinoma in-situ (LCIS). LCIS is not thought to be the direct precursor of the invasive component. The authors analyzed 15 cases of coexisting LCIS and IDC and found ductal carcinoma in situ (DCIS) in 12. The DCIS and IDC were of similar grade and located in the same area. Selected cases stained with E-cadherin demonstrated a different immunophenotype for the lobular and ductal lesions. These results support the notion that DCIS is the direct precursor of IDC occurring in patients with LCIS. PMID- 11484501 TI - Diagnostic pitfalls in surgical pathology--discovered by a review of malpractice claims. PMID- 11484502 TI - From histiocytosis X to Langerhans cell histiocytosis: a personal account. PMID- 11484503 TI - Necrotizing angiitis of the small intestine related to AA-amyloidosis: a novel association. AB - A 71-year-old man with intestinal pseudo-obstruction was found to have a diffusely thickened adynamic small bowel with AA-amyloid in submucosal vessels and muscularis propria, foreign body giant cell reaction to amyloid, and necrotizing angiitis. The mucosa was unremarkable. Immunostains demonstrated numerous CD68+ monocyte/macrophages and CD8+ T cells associated with the amyloid deposits. The patient had no evidence of systemic vasculitis and no underlying cause for AA-amyloidosis was identified. Necrotizing angiitis coexistent with amyloid angiopathy has been reported in brain and temporal arteries, but not in the gastrointestinal tract and not with AA-amyloid. The inflammatory cell infiltrates in this case are consistent with a foreign-body and/or cell-mediated immunologic reaction to AA-amyloid, although a role for these cells in amyloid formation cannot be excluded. PMID- 11484504 TI - Association of lymphangioleiomyomatosis (LAM) with endosalpingiosis in the retroperitoneal lymph nodes: report of two cases. AB - We report 2 patients in whom pulmonary lymphangioleiomyomatosis (LAM) affected the retroperitoneal lymph nodes and was associated with endosalpingiosis. These lesions were large, encapsulated masses with multiple cysts containing chylous fluid. Both were characterized by proliferating LAM cells that formed fascicles separated by slit-like channels. Some cysts were lined by ciliated epithelium resembling that of Fallopian tubes. Other cysts were lined either by flattened endothelial cells or by a mixture of these cells and epithelial cells. Many LAM cells gave a positive reaction with HMB-45 antibody. Most LAM cells in fascicles were reactive for alpha-smooth muscle actin and desmin. In 1 patient, many of the epithelial cells and some of the subjacent LAM cells were positive for estrogen and progesterone receptors. In conclusion, immunostaining with HMB-45 antibody and markers for smooth muscle cells can be helpful in the evaluation of problems in the differential diagnosis of lesions of extrapulmonary LAM, particularly those involving the genital system. PMID- 11484505 TI - Primary angiocentric T-cell intestinal lymphoma with Epstein-Barr virus in a 5 year-old boy. AB - A case of peripheral T-cell lymphoma affecting the small bowel of a 5-year-old boy is reported. The cells did not form a tumoral mass but infiltrated diffusely, arranged in an angiocentric pattern and associated with numerous ulcers, one of which perforated. Immunohistochemistry proved them to be CD45RO-, CD3-, and CD8 positive. CD20, CD4, and CD56 markers were negative. The presence of EBV in the lymphomatous cells was demonstrated by in-situ hybridization. Polymerase chain reaction study revealed T-cell receptor (TCR) gene rearrangement. Notably the hemophagocytic syndrome present on admission reverted after surgery. The authors are not aware of a previous report of intestinal T-cell lymphoma in a child. PMID- 11484506 TI - Combined carcinoid tumor and ganglioneuroblastoma of the lung: a case report. AB - Carcinoid tumor is a well-differentiated epithelial neuroendocrine neoplasm which is common in the lung. Ganglioneuroblastoma is a moderately differentiated nonepithelial neuroendocrine neoplasm which is very rare in the lung. Neuroendocrine tumors with epithelial and nonepithelial elements are rare in any site and have not been reported in the lung. This case is an example of a primary neuroendocrine lung tumor combining epithelial and non-epithelial components: carcinoid tumor and ganglioneuroblastoma. PMID- 11484507 TI - An evolutionary view of neuroendocrine cells and their tumors. PMID- 11484508 TI - Intratubular germ cell neoplasia: associated infertility and review of the diagnostic modalities. AB - The incidence of testicular neoplasia has increased, and its early detection has become a pressing clinical issue. The strong association between male subfertility and risk of testicular neoplasia is consistent with the existence of common pathogenetic factors. Most forms of testicular germ tumors are believed to stem from a common precursor, intratubular germ cell neoplasia (ITGCN), also known as testicular carcinoma in situ. Identification of ITGCN cells in testicular biopsies, however, is a diagnostic challenge and markers are sorely needed to assist in the accurate identification of the lesion. PMID- 11484509 TI - Immunoreactivity of p53, mdm2, and p21WAF1 in dedifferentiated liposarcoma: special emphasis on the distinct immunophenotype of the well-differentiated component. AB - Alteration of the p53/mdm2 pathway has been reported in the well-differentiated liposarcoma (WDLS)/dedifferentiated liposarcoma (DDLS) group. We investigated the immunoreactivity of p53, mdm2, and p21WAF1, along with the MIB-1-labeling index (MIB-1-LI) in 21 WDLS and 21 DDLS cases, to clarify the association of these markers with the morphologic changes and the biological factors responsible for the aggressiveness of DDLS. Within DDLS, p53 and p21WAF1 expression and mdm2 overexpression were significantly more prevalent in the dedifferentiated (DD) components than in the well-differentiated (WD) components. The mdm2 overexpression and p21WAF1 expression was significantly associated with sclerosing liposarcomas in both WDLS and the WD components of DDLS. There was no significant difference in the immunoreactivity of p53, mdm2, or p21WAF1 or MIB-1 LI between WDLS and the WD components of DDLS. An association was found between p53 expression and mdm2 overexpression in the WD group (comprising WDLS and WD components of DDLS) and in the DD group, significantly so in the WD group. Notably, this correlation was found in the subtype of sclerosing liposarcoma but not in that of lipoma-like liposarcoma. Within DDLS, the clinical outcome of the nonaccessible soft tissue (non-AST: comprising retroperitoneum and mediastinum) group was significantly worse than that of the accessible soft tissue (AST: comprising extremities, buttocks, axilla, and scrotum) group; however, the immunophenotypes of p53, mdm2, and p21WAF1 and the MIB-1-LI showed no correlation with survival in the AST group alone, in the non-AST group alone, or in the 2 together. This study suggests that the immunoreactivity of p53, mdm2, and p21WAF1 is associated with the morphologic changes, but not with the biological factors responsible for the aggressiveness of DDLS. PMID- 11484510 TI - Update on the management of acute severe pancreatitis. AB - Acute severe pancreatitis is an aggressive disease with a mortality rate of up to 30 percent. In recent years therapy has shifted away from early surgery to intensive medical care. This article focuses on several issues of the management of acute severe pancreatitis emphasising evidence from recent clinical trials and recommendations from recent consensus conferences. Since a correct assessment of the severity of the disease is mandatory as early as possible in the treatment, several multiple scoring factor systems and individual risk factors are explained. The indications and the optimal timing of ERCP are discussed. Prophylactic administration of antibiotics, intravenously or by means of a selective digestive decontamination scheme, seems to be beneficial in decreasing morbidity but not mortality. Adequate nutritional support, preferably achieved by enteral feeding, is an important component in the supportive therapy. Protease inhibitors and anti-secretory drugs have not proven to be of benefit in improving outcome. Immunomodulating substances like platelet activating antagonists are promising but further studies are necessary to confirm the results of the early studies. Finally, indications for surgery are discussed. PMID- 11484511 TI - Monitoring of erythropoiesis by the serum transferrin receptor and erythropoietin. AB - Virtually all cells have transferrin receptors (a transmembrane glycoprotein) on their surface but in a normal adult, 80% of them are in the erythroid marrow. Some of them are lost into the circulation where they can be measured by immuno assays. A direct and highly significant correlation exists between serum transferrin receptor level and erythron transferrin uptake in humans. The measurement of serum transferrin receptor has wide clinical applications for the quantitation of erythropoiesis. It can be used to study erythropoiesis in situations in which ferrokinetics is not acceptable such as pregnancy. It is particularly useful for serial studies, e.i., for monitoring the recovery of erythropoiesis after stem cell transplantation or after treatment with erythropoietin. Combined with the determination of serum erythropoietin, both evaluated in relation to the degree of anemia, they provide a physiological approach to the diagnosis of anemia. Thus, the simultaneous determination of hematocrit, reticulocytes, serum transferrin receptor and serum erythropoietin has high discriminatory value in distinguishing between a defect in erythroid proliferation, maturation or red cell survival. It is also particularly useful for detecting the presence of multiple mechanisms of anemia in the same patient. PMID- 11484512 TI - New insights into insulin resistance pathophysiology: how it affects glucose and lipid metabolism. AB - Insulin resistance is a metabolic state encountered in many physiological and physiopathological conditions. Its pivotal effect on glucose and lipid metabolism is increasingly recognised. Over the last few years, numerous molecular/intracellular pathways affected by this condition have been elucidated. The author reviews some recent findings underlying several major metabolic abnormalities. An overview of current and future therapeutics is also provided. PMID- 11484513 TI - Registration of interstitial lung diseases by 20 centres of respiratory medicine in Flanders. AB - We report the results of a prospective registration of the prevalence of interstitial lung diseases by 20 centres of respiratory medicine in Flanders from January 1992 till July 1996. The 20 centres registered a total of 362 cases. Sarcoidosis (31% with inclusion of stage I, or 22% without stage I), idiopathic pulmonary fibrosis (20%), hypersensitivity pneumonitis (13%) and unclassified forms of interstitial lung diseases (9%) were the most often diagnosed diseases. The mean age of the patients was 52 +/- 17 (SD) years, but ranged from 32 +/- 10 years in histiocytosis X to 70 +/- 13 years in drug induced interstitial lung diseases. The male to female ratio was 1.3 but ranged from 2.3 in interstitial lung diseases due to inhalation of inorganic materials to 0.8 in interstitial lung diseases due to connective tissue disease. The diagnosis was considered certain in 60%, probable in 30% and possible in 10%. In 50% of the cases the diagnosis was proven by biopsy and the most often used biopsy technique was transbronchiolo-alveolar biopsy (40% of biopsies) followed by open lung biopsy (32%). At diagnosis vital capacity was 82 +/- 22% pred. (ranging from 92 +/- 19% in sarcoidosis to 71 +/- 24% in idiopathic pulmonary fibrosis) and CO-diffusing capacity was 77 +/- 19% pred. (ranging from 77 +/- 19% in sarcoidosis to 41 +/- 13% in drug induced pneumonitis). The regional frequency of interstitial lung diseases in Flanders varied widely and was not proportional to the regional distribution of the 20 contributing centres. In conclusion this registration provides interesting information on the occurrence and presentation of the different forms of interstitial lung diseases, although it may still underestimate the real prevalence and incidence of interstitial lung diseases. PMID- 11484514 TI - Serum leptin concentration in peritoneal dialysis patients: determinants, longitudinal evolution and circadian rhythm. AB - The serum concentration of leptin, an hormone secreted by adipocytes, is increased in obese and chronic renal failure patients. To determine the influence of peritoneal dialysis (PD) therapy on serum leptin levels, we analyzed its concentration in 23 patients on PD for an average of 26.8 +/- 7.1 months and compared it to that of 18 patients with chronic renal failure (creatinine clearance: 49.1 +/- 8.8 ml/min) and of 35 healthy control subjects. Leptin level was also reevaluated in 11 PD patients 9.3 +/- 1 months after the initial analysis. Finally, circadian leptin production was determined in 4 patients on automated PD (APD). Serum leptin was significantly higher in PD (31.9 +/- 7.8 ng/ml) than in chronic renal failure patients (15.2 +/- 5.9 ng/ml) and in healthy control subjects (9.6 +/- 1.1 ng/ml). Serum leptin level was significantly correlated with BMI in all three groups (except in PD males) and with the percentage of fat mass in both male and female PD patients. It did not correlate in PD patients with serum albumin concentration, free fat mass, residual diuresis, time on PD and characteristics of peritoneal permeability. At the second determination, serum leptin level had significantly increased in the PD patients although their respective BMI and serum creatinine concentration had remained virtually unchanged. Finally, in APD patients, the highest leptin level was observed at 08.00 a.m. These results demonstrate that serum leptin level is increased in PD patients and that it progressively rises under PD therapy. The circadian leptin production is delayed in APD patients probably suggesting a negative effect of the nocturnal glucose load on the regulation of its secretion. PMID- 11484515 TI - Antimicrobial susceptibility of nontyphoidal Salmonella isolated from humans in Belgium. AB - Human nontyphoidal Salmonella infections are the primary cause of foodborne disease in developed countries, resulting in considerable morbidity and occasionally death, especially in immunocompromised patients. Strains of Salmonella that are resistant to antimicrobial agents have become a world-wide health problem. Fluoroquinolones are drugs of choice for treatment of human invasive salmonellosis, and have been useful for the treatment of infections caused by multi-resistant strains. However, strains resistant to ciprofloxacin have been noted. A random sample of 378 Salmonella strains of human origin was collected during 1998. Their susceptibility to 11 antimicrobial agents was determined by the agar dilution method according to NCCLS standards. In total, 38 serotypes were represented of which S. Enteritidis (20.4%), S. Typhimurium (20.4%), S. Hadar (9.0%), S. Brandenburg (7.9%), S. Infantis (7.7%), and S. Virchow (5.3%) were the most common. All strains were susceptible to ceftriaxone and ciprofloxacin. For nalidixic acid the rate of resistance was 19.0%. Of the 72 strains resistant to nalidixic acid, 31 were S. Hadar, and thus 91.2% (31/34) of the S. Hadar isolates showed resistance to nalidixic acid. Most of the S. Hadar strains were also resistant to ampicillin, tetracycline and sulphamethoxazole, and an elevated MIC50 (0.25 microgram/ml) and MIC90 (1 microgram/ml) was observed for ciprofloxacin. The high rate of resistance to nalidixic acid can be a first step towards the development of resistance to ciprofloxacin. PMID- 11484516 TI - Streptobacillus moniliformis: case report and review of the literature. AB - Streptobacillus moniliformis is one of the causes of rat bite fever. The clinical and microbiological diagnosis of this rare infectious disease is often quite problematic. Penicillin, in sufficiently high dosis, is the treatment of choice and leads to a prompt resolution of the infection, whereas lack of treatment leads to a 13% mortality rate. As far as we know, this paper reports the first human isolate in Belgium. PMID- 11484517 TI - Transfusion of polymerized bovine haemoglobin in a patient with sickle cell anaemia and severe allo-immunization: a case report. PMID- 11484518 TI - Revascularisation of fresh compared with demineralised bone grafts in rats. AB - Revascularisation of bone grafts is influenced by both the anatomical origin and the pre-implantation processing of the graft. We investigated the revascularisation by entrapment of 141Ce (cerium)-labelled microspheres in large, fresh and demineralised syngeneic grafts of predominantly cancellous (iliac bone) or cortical (tibial diaphysis) bone three weeks after heterotopic implantation in rats. The mean (SD) 141Ce deposition index (counts per minute (cpm) of mg recovered implant/cpm of mg host iliac bone) was higher in fresh iliac bone grafts, 0.98 (0.46) compared to that of demineralised iliac bone, 0.32 (0.20), p < 0.001, and fresh tibial bone grafts, 0.51 (0.27), p = 0.007. We found no significant difference in the mean 141Ce deposition index between fresh tibial bone grafts and demineralised tibial bone grafts, 0.35 (0.42), p = 0.4, or between demineralised tibial grafts and demineralised iliac bone grafts, p = 0.8. The results suggest that whereas fresh cancellous grafts are revascularised more completely than fresh cortical grafts, there is no difference in the revascularisation of demineralised cancellous and cortical grafts. In addition, fresh cancellous bone is revascularised more completely than demineralised cancellous bone, whereas there is no difference between fresh and demineralised cortical bone. PMID- 11484519 TI - Effect of high dose and low dose aspirin on survival of random pattern flaps in rats. AB - This paper studies which of the physiological effects of aspirin is responsible for increasing the survival of random flaps in rats, found in an earlier experiment. We wished to confirm that the antiaggregating--antithrombotic effect was responsible for the increased survival of flaps without microvascular anastomosis. Three groups of rats with standardised random pattern flaps were used. The first two were given aspirin 200 mg/kg (high dose, n = 27) or 40 mg/kg (low dose, n = 21) and the third (n = 28) acted as controls. The beneficial effects of aspirin were restricted to the high dose group. Since the low dose group also showed antiaggregation of platelets, but without the anti-inflammatory or vasodilatory effects, the results indicate that the antiaggregating effect alone was not responsible for the increased survival of the flaps. PMID- 11484520 TI - Disappearance of catecholamine fluorescence from the adrenergic nerves in arterial grafts in rats: an experimental fluorescence histochemical study. AB - The disappearance of catecholamine fluorescence from the noradrenaline-containing sympathetic nerve fibres after arterial transplantation was studied using a femoral artery graft sutured to rat carotid artery. Glyoxylic acid-induced fluorescence was used to demonstrate adrenergic nerves histochemically. At six hours the network of fibres had started to degenerate, and catecholamine fluorescence from the adrenergic nerves had almost completely disappeared within 24 hours of grafting. Control specimens from normal femoral arteries showed a dense network of fluorescent adrenergic nerves. Based on observations of the relatively rapid liberation of catecholamines from the degenerating adrenergic nerves, we suggest that catecholamines liberated from degenerating adrenergic nerves may have an important role in early vasospasm in microvascular and coronary bypass surgery. PMID- 11484521 TI - Delayed surgery does not reduce the interface strength between the surface of a bone fracture and a self-curing injectable hydroxyapatite (Norian SRS). AB - The shear strength of the interface between bone and an injectable calcium phosphate bone substitute that cures to form a carbonated apatite (Norian SRS) was measured and related to the time that elapsed between injury and surgery. Eleven rabbits had a 3 mm drill hole made in the retropatellar aspect of the distal femur. After one week, the rabbits were reoperated on and the hole filled with Norian SRS after it had been cleaned gently with a sponge. During the same procedure a similar hole was burred in the opposite femur and filled with Norian SRS after a few minutes ("immediate injection"). Four other rabbits had only one side operated on with immediate injection of Norian SRS to compare with the opposite untreated side. The rabbits were killed four days after the injection of Norian SRS. The femurs were prepared and sawed perpendicularly to the burr channels to produce discs 3.5 mm thick. A push out test of the Norian SRS plug within the bone disc was done to measure the force at failure. In the immediately injected specimens the failure occurred at a mean of 28 N (range 5-57) compared with 42 N (range 25-65) in the specimens injected after a one week delay. There was no significant difference between delayed and immediate treatment (95% confidence interval -5 to 133). The histological examination showed that 9 of the 12 specimens had Norian SRS still adherent to half or more of the circumference of the hole. This indicates that the failure occurred in the Norian SRS rather than in the bone or at the interface. PMID- 11484522 TI - Neuronal loss after transsection of the facial nerve: a morphological and neurophysiological study in monkeys. AB - Functional recovery after nerve lesions seems to depend on peripheral as well as central factors. To investigate the central neuronal loss after transsection of a pure motor nerve, the middle branch of the facial nerve on one side was transsected and immediately repaired microsurgically by epineural suturing. After a period of 6-15 months, a quantitative neurophysiological recording was made to estimate muscle response. A nerve tracer was injected into the mimic muscles innervated by the nerve to label the surviving motor neurons within the facial nucleus. The opposite side was used as the control in all cases. After the regenerative period, a mean loss of 15% of the total cell number was observed within the facial nucleus compared with the opposite side. The cell loss comprised all types of neurons. This amount of neuronal loss was followed by an even greater loss of muscle response when a quantitative neurophysiological recording was made after nerve regeneration. The results are discussed in relation to loss of nerve elements after nerve lesions and its effect on functional recovery. PMID- 11484523 TI - Immunohistochemical study of clinical skin-penetrating titanium implants for orthopaedic prostheses compared with implants in the craniofacial area. AB - The technique of using osseointegrated bone-anchored percutaneous titanium implants in the rehabilitation of patients with amputated limbs has recently been introduced at the Branemark Osseointegration Centre in Goteborg. The new method is based on an implantation technique that is well-established for anchorage of bone-conductive hearing aids and craniofacial prostheses. The soft tissues around skin-penetrating titanium implants used to anchor orthopaedic prostheses were evaluated histochemically using quantitative analyses. The implants had been in clinical use for between 6 to 24 months. The number of inflammatory cells was higher in the area close to the interface than in the area distant from the skin penetrating site, and higher than in the corresponding controls. The current data was also compared with corresponding data from tissue around skin-penetrating craniofacial implants. The number of inflammatory cells was lower in the orthopaedic samples than in the craniofacial specimens. We conclude that skin penetration of orthopaedic implants is as safe as when titanium implants are used for craniofacial rehabilitation, which is a clinically well-established procedure. PMID- 11484524 TI - Expander elements in craniofacial surgery: an experimental study in rabbits. AB - With the inception of craniofacial surgery elaborate surgical procedures were introduced for the treatment of craniosynostosis. Recently the use of implantable springs to aid simple strip craniectomies for these conditions has been described. Having shown the feasibility of using springs for dynamic skull reshaping, several questions remain to be answered, all of them about how to control the spring action clinically. One of the most important questions concerns force. The purpose of this study was to evaluate experimentally the effect of two springs of different strengths on the growth of rabbit calvaria after strip craniectomy. Thirty-two 6-week-old rabbits were randomised into one of four groups: sham-operation where only amalgam markers were inserted after subperiostal dissection; strip craniectomy of the sagittal suture, no expansion; strip craniectomy of the sagittal suture and insertion of an expander element made of titanium molybdenum alloy (TMA); and strip craniectomy sagittal suture and insertion of an expander element made of stainless steel (SS). SS springs delivered a range of forces from 2.42-2.18 N, whereas the TMA expander elements delivered a range from 1.39-1.09 N. The parietal bone marker separation after 12 weeks was 4.9 mm in the TMA group and 7.4 mm in the SS group. This resulted in a significant increase of the calvarian height compared with control groups. Histological examination showed intramembranous bone formation in the bone gap in all groups. PMID- 11484525 TI - Long-term follow-up of dynamic cranioplasty for brachycephaly--non-syndromal bicoronal synostosis. AB - We followed up 10 patients whose non-syndromal bicoronal synostosis had been operated on with a dynamic cranioplasty technique developed by this craniofacial unit in 1992. With this technique, the growth of the brain is redirected in an anteroposterior direction as wire-mediated compression and restraint are exerted on the transverse and vertical dimensions of the skull. The mean operating time was 160 minutes (range 120-275) and mean stay in the intensive care unit was 36 hours (range 23-58). There was no operative mortality and few complications. The surgical results were assessed objectively by analysis of cephalometric tracings. The mean (SD) cephalic index was 87.6 (4.9) preoperatively and 77.7 (1.8) postoperatively (p = 0.001). The modified Whitaker scale was used as a subjective outcome measurement, and nine patients were classified as Whitaker grade 1 (no additional surgery). One patient required additional intracranial surgery. A questionnaire was sent to all families to obtain an additional subjective measurement of outcome. Parents' satisfaction was high. We conclude that dynamic cranioplasty is a safe and efficient operation for treatment of brachycephaly. PMID- 11484526 TI - Skeletal analysis of craniofacial deformities in brachycephaly: comparison with craniofacial deformities in plagiocephaly. AB - The deformities of craniofacial bones in brachycephaly (n = 2) and plagiocephaly (n = 2) were compared using three-dimensional skull replicas. In brachycephaly the bilateral deformities of the cranial base were similar to the unilateral deformities on the affected side in plagiocephaly. Shortening of the anterior middle cranial fossas and expansion of the middle cranial fossas suggest downward and anterior displacement of both temporomandibular (TM) joints resulting in the underdevelopment of the middle-inferior facial bones, though this is less conspicuous in brachycephaly for three reasons. The most important one is that the patients have an imposing facial morphology characterised by high, protruding facial bones, as found on the affected side in plagiocephaly. Secondly, the frontal bones are flattened and positioned posteriorly in the anteroposterior direction, which also helps to mask the underdevelopment of the middle-inferior facial bones. Finally, as bony deformities are symmetrical in brachycephaly, they are not as obvious as they would be if they were unilateral. PMID- 11484527 TI - Identification of children at high anaesthetic risk at the time of primary palatoplasty. AB - Closure of the palate is an operation with a higher risk of postoperative hypoxaemia than other plastic surgical procedures. An increase in anaesthetic complications was noticed among our children with a cleft palate who were operated on with a palatoplasty, and this was further investigated. The records of 154 patients operated on between 1979 and 1996 were eligible for the study. A total of 128 had isolated cleft palate (ICP), 19 had Pierre Robin syndrome (PRS), and seven had other identified syndromes (IS). The results showed that the risk of anaesthetic complications was four times greater when the operation was done when the child was less than 1 year old, and there was a sixfold increase when a more elaborate velopharyngoplasty technique was used. Children with Pierre Robin syndrome were further classified according to the degree of neonatal difficulties. The 12 children with most severe problems were at increased risk. When the diagnostic criteria as well as the type of operation were taken into account the incidence of anaesthetic complications was 7/11, which was the highest risk encountered in the present study. When the first operation on the palate in children with Pierre Robin syndrome is planned, it should be based on a classification of neonatal problems and an investigation of their clinical condition. In children at risk the palatoplasty procedure should be postponed until the age of 12-18 months. PMID- 11484528 TI - The lateral thoracodorsal flap in breast reconstruction: a long-term follow up study. AB - Fifty-four patients were studied a mean of five years after their breasts had been reconstructed between 1984 and 1990 using the lateral thoracodorsal (LTD) flap in combination with either a thin shell, non-low-bleed (n = 35) or a thick shell, low bleed (n = 19) silicone gel implant. The rate of capsular contracture (Baker III-IV) was 11% in the first group and 10% in the second according to a modified Baker classification. Open capsulotomy was common in both groups of patients (15/35 in the first group and 13/19 in the second). Investigation by applanation tonometry of the capsular contracture agreed with the modified Baker classification. The cosmetic results were evaluated clinically and from photographs. Best scores were recorded for scars and ptosis in both evaluations. There were no significant differences between the general cosmetic results in the two groups. The patients graded their estimations of the final outcome of their breast reconstruction on a 10-point visual analogue scale (VAS); the mean for the first group was 8.7 and for the second 9.2. None of the patients regretted her operation and they would all recommend the procedure to another patient. PMID- 11484529 TI - Use of tissue expansion in revision of unhealed below-knee amputation stumps. AB - When local tissue is insufficient for the revision of unhealed below-knee stumps tissue expansion offers an interesting alternative for local coverage. We used this method in seven patients (five men, two women; mean age 30 years) who had had below-knee amputations, six of them after injury to a healthy limb and one for purpura fulminans. Ten tissue expanders were inflated slowly and intermittently either weekly or twice weekly depending on the patients. The mean expansion period was 92 days. Mean hospital stay for the two operations was 5.8 (range 4-9) and 7.6 (range 6-10) days, respectively. Using subjective and objective criteria, functional outcome was excellent in five patients and good in one. Expansion failed in one because of infection. Expanded skin flaps allow good cover with a minimal scar area in appropriate cases, while preserving the skin sensitivity and length of the tibial shaft. PMID- 11484530 TI - The treatment of port wine stains with a dye laser: a study of 644 patients. AB - Between June 1989 and December 1996, the flashlamp pumped pulsed dye laser was used to treat port wine stains (PWS) in 644 patients, age range 3 months-93 years (mean 21). The efficacy of the treatment was assessed after more than one year of follow-up. Each factor that might affect the efficacy was then evaluated statistically. Broad lesions required more laser treatments than narrow lesions and clearing tended to start from the periphery of the lesion, indicating the three-dimensional depth of the broader PWS. Patients who had been given previous treatments such as argon laser required about two more laser treatments than those who had not, but there were no clear differences in the efficacy of dye laser treatment between the two groups. PMID- 11484531 TI - Augmentation mammaplasty in male-to-female trans-sexuals: facts and figures from Amsterdam. AB - In male-to-female trans-sexuals in whom the hormonal treatment has not resulted in sufficient growth of the breasts, augmentation mammaplasty may be considered. Mammaplasty is performed in two-thirds of our patients who have had vaginoplasties. We present our experience of 201 patients operated on at the Academisch Ziekenhuis Vrije Universiteit in Amsterdam up to 1997. The patients' mean age was 34 years (range 17-76) and the mean follow-up was four years (range 2 weeks to 16 years). In keeping with our protocol, most patients had had oestrogen therapy and a successful real-life experience for a minimum of 1.5 years. The mean size of implants has nearly doubled over the years. Male-to female trans-sexuals often request large breasts and repeated augmentation mammaplasties were required, mainly in cases where the result of initial augmentation was not outspoken. PMID- 11484532 TI - Axial bilobed flap based on the median and medial superficial sural arteries: a case report. AB - The anatomy of the sural artery has been adequately described, but its applications are constantly evolving. Based on pre-existing anatomical studies, an axial bilobed flap based on the median and medial superficial sural arteries was developed and used in one patient for the reconstruction of a contracture of the popliteal area. The main advantages of this flap are that the donor site is closed primarily, it provides an adequate amount of healthy and pliable skin with an excellent blood supply and no chance of contraction, and it is an easy technique. PMID- 11484533 TI - Pregnancy and tram-flap breast reconstruction after mastectomy: a case report. AB - A 38-year-old patient had a right radical (Patey) mastectomy for an infiltrating ductal carcinoma followed by chemotherapy and, one year later, a TRAM flap breast reconstruction. She was given clear and exhaustive information about the possible consequences of pregnancy, but despite this she became pregnant four months after the reconstruction. The pregnancy was taken to term despite the appearance at four months of a slow-growing abdominal hernia. In the light of our experience and that of others, we recommend an interval of at least 12 months between breast reconstruction with a TRAM flap and pregnancy. PMID- 11484534 TI - Radial nerve entrapment in the upper arm as a cause of lateral arm pain: a report of four cases. AB - Four patients with no history of trauma presented with lateral arm pain, local tenderness, and a tingling sensation at the distal end of the arm when the radial nerve was percussed in the mid-third of the upper arm (Tinel's sign), but no clinical or subjective signs of muscular weakness. They were treated by decompression of the radial nerve in the fibrous canal proximal to the lateral intermuscular septum. Three of the patients had a complete or pronounced reduction in pain, while the fourth had only a slight improvement. Non-traumatic radial nerve entrapment in the upper arm may be the cause of lateral arm pain without clinical signs of muscular weakness. PMID- 11484535 TI - Congenital defect in the abdominal wall in an adult: a case report. AB - We describe a rare case of a woman aged 38 who presented with an untreated omphalocele in association with a duplication of the vena cava and malposition of the right kidney. An aesthetic and functional reconstruction of the abdomen was achieved with local flaps and the creation of a neoumbilicus. PMID- 11484536 TI - Matters of life and death. PMID- 11484537 TI - And now, ethics for sale? PMID- 11484538 TI - Trials and errors: a hospital takes a hit. PMID- 11484539 TI - Are boys the weaker sex? PMID- 11484540 TI - Reports of thyroid drug's demise were exaggerated. PMID- 11484541 TI - Nutrient overload at breakfast. PMID- 11484542 TI - Drug firms have workers 'under siege'. PMID- 11484543 TI - 'A handy interviewing tool' fell short. PMID- 11484544 TI - No untrained workers may use our service. PMID- 11484545 TI - Turn it down! PMID- 11484546 TI - The road to process safety success. PMID- 11484547 TI - Wasting away. PMID- 11484548 TI - The dangers of one-dimensional RM. PMID- 11484549 TI - Disheartening losses in U.S. steel industry. PMID- 11484550 TI - Hand dominance and balance reactions in the workplace. PMID- 11484551 TI - Of comp & communication. PMID- 11484552 TI - Planning and organizing a safety program. PMID- 11484553 TI - Plain-vanilla compliance. PMID- 11484554 TI - Worker sent to the hospital after heat stress event. PMID- 11484555 TI - New software: did we get what we expected? PMID- 11484556 TI - For the record. PMID- 11484557 TI - OSHA's revised pathogens standard. PMID- 11484558 TI - KISS (keep it simple, stupid)--to save a life. PMID- 11484559 TI - Volatile protection. PMID- 11484560 TI - Falls under control. PMID- 11484561 TI - Anyone at any height.... PMID- 11484562 TI - Safety in numbers. Changing your work culture will benefit your bottom line. PMID- 11484563 TI - When the mercury soars. PMID- 11484564 TI - Winning with wellness. PMID- 11484565 TI - Continuing prosthodontic research. PMID- 11484566 TI - Clinical longevity of removable partial dentures retained by telescopic crowns: outcome of the double crown with clearance fit. AB - PURPOSE: It was the aim of this study to investigate the long-term success of a telescopic crown system that can be used for both rigid and resilient support and to evaluate by means of a literature review whether the use of resilient support may be advantageous compared to other double crown systems when the restoration is placed on only a few remaining teeth. MATERIALS AND METHODS: Patient records were used to evaluate 125 dentures (with 460 abutment teeth) based on the Marburg double crown system. The loss of abutment teeth, endodontic treatment, and fracture of the metallic framework were investigated with regard to the different types of denture support. RESULTS: The probability that a patient would have kept all abutment teeth was 84% after 5 years and 66% after 10 years. No significant differences were found for the two groups "resilient support" (< or = three abutment teeth) and "rigid support" (> or = four abutment teeth). For abutment teeth with a double crown with clearance fit, the risk of loss was 4% after 5 years and 15% after 10 years for rigid support, and 10% and 24%, respectively, for resilient support. The risk of endodontic treatment was 7% after 5 years and 9% after 10 years for rigid support, and 3% and 7%, respectively, for resilient support. None of the denture frameworks showed a fracture during the observation period. CONCLUSION: Removable partial dentures retained by double crowns with clearance fit and constructed without major or minor connectors provide good clinical longevity. The survival rates of abutment teeth were comparable to those reported in the literature for other double crown systems. There was no significant increase of the risk of abutment loss when the restoration was placed on three or fewer remaining teeth and the concept of resilient support was applied. PMID- 11484567 TI - Some mechanical properties of a highly cross-linked, microwave-polymerized, injection-molded denture base polymer. AB - PURPOSE: The impact strength and the flexural properties of denture base materials are of importance in predicting their clinical performance upon sudden loading. This study compares the impact and transverse strengths and the flexural modulus of three denture base polymers. MATERIALS AND METHODS: The investigation included a relatively new microwave-polymerized polyurethane-based denture material processed by an injection-molding technique, a conventional microwave polymerized denture material, and a heat-polymerized compression-molded poly(methyl methacrylate) (PMMA) denture material. Impact strength was determined using a Charpy-type impact tester. The transverse strength and the flexural modulus were assessed with a three-point bending test. The results were subjected to statistical analysis using a one-way analysis of variance and the Scheffe test for comparison. RESULTS: The impact strength of the microwave-polymerized injection-molded polymer was 6.3 kl/m2, while its flexural strength was 66.2 MPa. These values were lower than those shown by the two compression-molded PMMA-based polymers. The differences were statistically significant. The flexural modulus of the new denture material was 2,832 MPa, which was higher than the conventional heat-polymerized polymer but was comparable to the other microwave-polymerized PMMA-based polymer. The difference in the flexural modulus was statistically significant. CONCLUSION: In terms of the impact and flexural strengths, the new microwave-polymerized, injection-molded, polyurethane-based polymer offered no advantage over the existing heat- and microwave-polymerized PMMA-based denture base polymers. However, it has a rigidity comparable to that of the microwave polymerized PMMA polymer. PMID- 11484568 TI - Clinical survey of acrylic resin removable denture repairs with glass-fiber reinforcement. AB - PURPOSE: The aim of this study was to evaluate clinical usefulness and durability of continuous glass-fiber reinforcement in repair of acrylic resin removable dentures. MATERIALS AND METHODS: Fractured removable dentures without reinforcement, with conventional metal-wire reinforcement, or with mesh reinforcement were collected from two dental schools in Finland. The total number of dentures was 51 and the number of patients was 48. During the repair, the dentures were reinforced with a polymer-preimpregnated E-glass fiber at the region of the fracture. The fibers were used as partial fiber reinforcement, i.e., only the weakest part of the denture was reinforced. Follow-up time varied from 4 months to 4.1 years. After the follow-up period, possible fractures and discoloring were visually inspected. Possible irritation of oral mucosa by glass fibers and the general shape of the denture were also evaluated. RESULTS: In 88% of the cases, there was no need for adjustment at the region of partial fiber reinforcement, and the clinical condition of the dentures was good. Glass fibers did not irritate the oral mucosa. In the case of refracture or hairline fracture, positioning of the partial fiber reinforcement was incorrect or the reinforcement had been used incorrectly (the wetting of the reinforcement with denture base resin was inadequate). CONCLUSION: Polymer-preimpregnated partial fiber reinforcement seems to be useful in eliminating fractures of acrylic resin removable dentures. However, this study emphasizes the importance of correct positioning and accurate laboratory technique when partial fiber reinforcement is used. PMID- 11484569 TI - Effect of priming time on tensile bond strength to bovine teeth and morphologic structure of interfaces created by self-etching primers. AB - PURPOSE: The purpose of this study was to investigate the effect of priming time on bovine enamel and dentin adhesion promoted by self-etching primer systems. MATERIALS AND METHODS: Two commercial self-etching primer systems, Clearfil SE Bond (SE) and Unifil Bond (UB), were used. Bovine enamel and dentin were treated with each system with various priming times (5, 20, and 60 seconds), and the tensile bond strength was measured. Scanning electron microscopic (SEM) observation was also performed to examine the effect of priming time on the morphology of the resin-tooth interface. RESULTS: The tensile bond strength to enamel ranged from 10.6 to 14.3 MPa, and no statistically significant difference was detected among products or priming times. To dentin, UB showed the statistically significantly lowest tensile bond strength when primed for 5 seconds, 6.9 +/- 1.1 MPa, but there was no significant difference between the groups primed for 20 and 60 seconds, 13.5 +/- 4.1 MPa and 13.8 +/- 4.6 MPa, respectively. SE created tensile bond strengths ranging from 13.9 to 15.7 MPa and showed no effect of priming time on dentin adhesion. SEM observation revealed that resin penetrated into both enamel and dentin more deeply with extension of priming time. CONCLUSION: Priming times longer than those recommended by the manufacturers did not influence the tensile bond strength to enamel and dentin when using two commercial self-etching primers. In contrast, a shortened priming time, 5 seconds, carried a risk of decreasing dentin adhesion, although it had no adverse effect on enamel adhesion. PMID- 11484570 TI - Fracture resistance of lithium disilicate-, alumina-, and zirconia-based three unit fixed partial dentures: a laboratory study. AB - PURPOSE: The purpose of this study was to determine the fracture resistance of three-unit fixed partial dentures (FPD) made of new core ceramics. MATERIALS AND METHODS: A base metal three-unit master FPD model with a maxillary premolar and molar abutment was made. Tooth preparation showed 0.8-mm circumferential and 1.5 mm occlusal reduction and a chamfer margin design. FPDs were constructed with a uniform 0.8-mm-thick core ceramic and a porcelain veneer layer. In-Ceram Alumina, In-Ceram Zirconia, and DC-Zirkon core ceramics were machined by a computer-aided design/manufacturing system, whereas IPS Empress 2 core ceramic was indirectly built up using the fabrication technology of waxing and heat pressing. FPDs of IPS Empress were heat pressed as complete restorations without core material. To ensure standardized dimensions, the FPDs were controlled at different points. All FPDs were cemented with ZnPO4 on the master model and loaded on a universal testing machine until failure. The failure load and mode of failure were recorded. RESULTS: The highest failure loads, exceeding 2,000 N, were associated with FPDs of DC-Zirkon. FPDs of IPS Empress and In-Ceram Alumina showed the lowest failure loads, below 1,000 N, whereas intermediate values were observed for FPDs of IPS Empress 2 and In-Ceram Zirconia. Differences in mean values were statistically significant. CONCLUSION: The high fracture resistance evaluated for FPDs made of DC-Zirkon underscores the remarkable mechanical properties of high performance ceramic, which could be useful for highly loaded all-ceramic restorations, especially in the molar region. PMID- 11484571 TI - Effect of surface treatment on fatigue life of postceramic soldered joints. AB - PURPOSE: This research investigated the effect of surface treatment on the fatigue life of metal-ceramic postsoldering. MATERIALS AND METHODS: Twenty cylindric specimens were cast in a metal-ceramic alloy. All specimens received appropriate heat treatment simulating ceramic application, although no porcelain was applied. Each specimen was cut in half to form two half specimens. The 40 half specimens were randomly assigned to one of the four treatment groups, which differed in the type of surface treatment performed on one end of each half specimen (joint surface) prior to soldering: (1) aluminous oxide pink stone; (2) 50-micron aluminum oxide sandblasting; (3) brown rubber point; and (4) gray silicone wheel followed by pink silicone wheel. All surface treatments were performed for 30 seconds. The half specimens were then steam cleaned, aligned, indexed, and oven soldered with #650 postceramic solder. The soldering of two half specimens formed a complete test specimen, and a total of 20 postceramic soldered specimens were prepared. Following soldering, a 241.1 MPa fatigue stress was applied to each solder joint during specimen testing. The test variable was the number of stress cycles required to fail each specimen. RESULTS: All specimens failed adhesively at the joint interface between the solder and parent metal. There were significant differences in the number of stress cycles to failure between groups 1 and 2, groups 1 and 4, and between groups 2 and 3. CONCLUSION: The load cycle to failure for postceramic soldered joints was affected by the metal surface treatment. PMID- 11484572 TI - Restoring endodontically treated teeth: a survey of current opinions among board certified prosthodontists and general dental practitioners in Sweden. AB - PURPOSE: The main aim was to investigate the current opinions among general dental practitioners and board-certified prosthodontists in Sweden on how to restore root-filled teeth. MATERIALS AND METHODS: A questionnaire containing 31 multiple-choice questions was mailed to 892 general practitioners and 150 board certified prosthodontists in Sweden. The questions asked for information on when the clinicians used posts in endodontically treated teeth and what kind of clinical procedure they used. They were also asked if they believed that a post strengthened a root-filled tooth. RESULTS: Sixty percent of the general practitioners and 67% of the prosthodontists returned the questionnaire. Twenty nine percent of the responding general practitioners and 17% of the prosthodontists were of the opinion that a post reinforces a root-filled tooth. Only a few clinicians used posts "always" or "most of the time" when restoring endodontically treated teeth with fillings, while the vast majority used posts when restoring such teeth with crowns or fixed partial dentures. In both groups of dentists, cast posts were most commonly used. Despite the present knowledge that parallel-sided posts have a significantly higher success rate than tapered cast posts, only a minority of Swedish dentists use parallel-sided posts. CONCLUSION: In contrast to the results of several current studies, a high proportion of both general practitioners and prosthodontists believe that a post reinforces an endodontically treated tooth. This is one probable explanation for the almost ubiquitous application of posts when teeth are restored with crowns or fixed partial dentures. PMID- 11484573 TI - Wettability of visible light-curing denture lining materials. AB - PURPOSE: Wetting characteristics of denture lining materials indicate the degree of salivary lubricating effect, which promotes denture retention and patient comfort. This in vitro study investigated the wettability of ten commercially available visible light-cured denture lining materials. MATERIALS AND METHODS: Ten soft and hard visible light-curing materials, one autopolymerized hard lining material, and one autopolymerized denture base material were evaluated for wettability. Wettability was estimated by measuring the equilibrium and hysteresis contact angles using the dynamic contact angle analysis, or Wilhelmy, technique. RESULTS: The equilibrium contact angle ranged from 59.9 to 77.3 degrees, and contact angle hysteresis ranged from 14.7 to 30.6 degrees. CONCLUSION: Visible light-curing lining materials exhibit wetting properties similar to the conventional hard lining and denture base materials. PMID- 11484574 TI - Interchangeability of two semiadjustable articulators. AB - PURPOSE: Interchangeability between articulators allows mounted casts to be transferred accurately from one articulator to another. The purpose of this study was to evaluate the interchangeability of 109 new, unused interchangeable articulators in the centric position. MATERIALS AND METHODS: The interchangeability of 38 unused Whip Mix model #2240 and 71 unused model #3040 semiadjustable articulators was evaluated using the Whip Mix #2245 check system as the standard. Vertical discrepancies between the check system cylinders were also measured. RESULTS: Within the standard for interchangeability established by the Whip Mix #2245 check system (< 94 microns horizontal discrepancy), 101 (93%) of 109 articulators were interchangeable (34 were model #2240 and 67 were model #3040). Overall, the amount of vertical space between the check system cylinders was < or = 52 microns in 99.4% of the model #2240 articulators and 99.7% of the #3040 articulators. CONCLUSION: New, unused Whip Mix model #2240 and #3040 articulators are potentially interchangeable in the centric position. PMID- 11484575 TI - Fracture behavior of human mandibular incisors following endodontic treatment and porcelain veneer restoration. AB - PURPOSE: Because of existing controversy, the present study investigated the individual and combined effects of endodontic treatment and porcelain veneer restoration on the fracture behavior of human mandibular incisors. MATERIALS AND METHODS: Forty extracted intact human mandibular incisors were assigned to four groups of ten with a similar range of labiolingual widths at the cementoenamel junctions. Group A consisted of intact teeth; group B consisted of endodontically treated teeth; group C teeth were restored with labial porcelain veneers; and those of group D were endodontically treated and had labial porcelain veneers. All teeth were subjected to a slow continuous loading test at 30 degrees to the long axis of the teeth and 1 mm below the incisal edge on the labial side. RESULTS: Fracture forces were 415 +/- 220 N, 370 +/- 89 N, 420 +/- 128 N, and 448 +/- 156 N for groups A, B, C, and D, respectively. Root fracture was the most common mode of failure. There were no statistically significant differences between the groups in terms of fracture forces and modes of failure. CONCLUSION: Human mandibular incisors with endodontic treatment and/or porcelain veneer restorations were able to withstand the same magnitude of oblique loading as intact teeth. Endodontic treatment and/or porcelain veneer restoration did not affect the mode of failure of mandibular incisors. PMID- 11484576 TI - Interobserver variability in assessment of signs of TMD. AB - PURPOSE: The aim of this report was to study the ability of examiners to measure reliably the clinical signs of temporomandibular disorders (TMD). Four examiners participated in this study of 11 TMD patients and 25 nonpatients. MATERIALS AND METHODS: Vertical and lateral excursions of the jaw were measured using a millimeter ruler. Joint sounds during vertical jaw movements were assessed using digital palpation. The reliability of delivering appropriate degrees of digital pressure to assess masticatory muscle pain was assessed using a manometer after training examiners to exert specified pressures. RESULTS: Intraclass correlation coefficients for the measurement of vertical and protrusive jaw movements were > or = 0.87, which was considered excellent. The intraclass correlation coefficient for measurements of left and right lateral jaw excursions varied between 0.73 and 0.85, which was considered acceptable. The interobserver agreement for detecting the joint sounds showed overall agreement across examiners of 78%. Kappa for every possible pair of examiners varied between .52 and .86 (median .75, interquartile range .18). Reliability for diagnostic categories from the Helkimo index and Research Diagnostic Criteria for Temporomandibular Disorders involving joint noises showed modest reliability. CONCLUSION: Point estimates and measures of spread for reliability measures of single clinical TMD signs as well as combinations of signs into diagnostic categories from the Helkimo index and Research Diagnostic Criteria for Temporomandibular Disorders involving joint noises were sufficient in a group of four examiners. PMID- 11484577 TI - Shear bond strength of denture reline polymers to denture base polymers. AB - PURPOSE: This study was undertaken to characterize the shear bond strength established between four denture base polymers and four denture reline polymers. MATERIALS AND METHODS: Cylindric columns of denture reline polymers were bonded to columns of denture base polymers. Specimens were immersed in water for 4 months and then thermocycled. The strength at which the bond failed under shear stress was recorded. RESULTS: Significant differences in bond strength existed among the specimens because of the denture base polymer variable, the denture reline polymer variable, and their interaction (P < .05). A light-activated denture base polymer (Triad) bonded adequately with a light-activated reline polymer (Triad) but less well with the other reline polymers tested. The bond strength established between some denture base polymers and a different light activated reline polymer (Rebaron LC) was relatively low. CONCLUSION: The type of denture base polymer and denture reline polymer affected the shear bond strength between them. PMID- 11484578 TI - On a new method to assess the accuracy of a CAD program. AB - PURPOSE: This study was initiated with the purpose of developing and evaluating a system for measuring the magnitude of the variation between a computer-aided design (CAD) object created on the computer screen and a replicated object produced by computer-aided manufacturing (CAM). MATERIALS AND METHODS: Controlled geometric forms, a square and a cone, were designed in a CAD program, and measuring distances were selected. The CAD data were transmitted to CAM data, and objects were produced. The CAD/CAM process was the Procera system, and the holder system permitted the objects to be ground into cross sections, similar to the visualization in the CAD program. Five objects of each shape were produced and measured by two operators two times. Three operators measured one of the objects 30 times. RESULTS: Observed values were greater than the true value. For all objects, the systematic error was at most 15.5 microns. Interoperator difference was small. The variation because of measurement error was greater for the square object compared to the cone. However, the variation because of object was higher for the cone object than for the square. The total standard deviation was 7.7 microns. Thus, the total random error caused by object variation and measurement error was in approximately 95% of all measurements less than 15 microns. CONCLUSION: There are no differences in the measurement data derived from this method and actual measurement data from an object created by the computer-aided dental design program. The method has high validity and reliability, i.e., high accuracy. PMID- 11484579 TI - Resistance to staining, flexural strength, and chemical solubility of core porcelains for all-ceramic crowns. AB - PURPOSE: The increased demand for tooth-colored restorations has prompted the use of ceramics in areas that are subject to masticatory stresses. To maximize the strength of these restorations, manufacturers and clinicians advocate placement of core materials in lieu of veneering materials in areas that are more susceptible to fracture. The objectives of this study were to determine the: resistance to staining of three core porcelains used for all-ceramic restorations, Procera, IPS Empress, and In-Ceram, through the use of colorimetry and visual observation; flexural strength of these porcelains under a three-point bend test; and chemical solubility in a controlled environment. MATERIALS AND METHODS: L*a*b* values were obtained for each specimen before and after immersion in a saturated solution of methylene blue in ethanol for 24 hours. Visual observation was also performed to ascertain color differences before and after staining. A three-point bend test was used to determine flexural strength. A reflux-condenser type, three-piece extraction apparatus was used with 4% acetic acid solution for 16 hours to determine solubility. Each sample was weighed before and after the reflux procedure to ascertain percentage weight loss. RESULTS: The mean delta E obtained for IPS Empress was 14.5; In-Ceram was 9.2, and Procera was 9.0. Flexural strength obtained for IPS Empress, In-Ceram, and Procera was 176.9 MPa, 323.4 MPa, and 464.3 MPa, respectively. Weight loss of IPS Empress, In-Ceram, and Procera was 0.056%, 0.734%, and 0.003%, respectively. CONCLUSION: IPS Empress showed the least resistance to staining. IPS Empress had the lowest flexural strength, while Procera had the highest. In-Ceram demonstrated the highest chemical solubility. PMID- 11484580 TI - Periodontal conditions among the old elderly: five-year longitudinal study. AB - In the past three decades, there has been a significant rise in the number of old elderly (85+ years old) in Finland, and more of these individuals are retaining their natural teeth. Numerous cross-sectional studies have reported on the periodontal health of the elderly (aged 75+), but very few long-term follow-ups have been reported. This study forms a part of the population-based Helsinki Aging Study (HAS) and compares the periodontal health status and the treatment needs at baseline with those of the same population five years later. The baseline study, in 1990-91, examined the dentate elderly born in 1904, 1909, and 1914, living in Helsinki, Finland (n = 196). The follow-up study was completed in 1995-96 (n = 73). Periodontal status was recorded by means of the Community Periodontal Index of Treatment Needs (CPITN) in 175 dentate subjects (55 males and 120 females) who met the criteria at baseline, and in 57 dentate elderly (17 males and 40 females), aged 81, 86, and 91 years, who remained at follow-up. Among the 57 dentate elderly who participated in both baseline and follow-up examinations, the mean number of teeth decreased from 15.9 to 15.1, and the mean number of remaining sextants from 4.2 to 3.7. There were minor changes in the periodontal health status during this five-year period, with an increase in code 2 (from 43% to 58%) and a decrease in code 3 (from 38% to 25%). Nevertheless, the overall treatment needs remained unchanged. It can be concluded that the periodontal health of the elderly had remained stable for 5 years, and almost no change was observed in their treatment needs. Therefore, periodontal disease in the elderly who are relatively healthy is not caused by the aging process. PMID- 11484581 TI - Perception of dry mouth in a sample of community-dwelling older adults in Japan. AB - The purpose of this study was to examine the prevalence of perceived dry mouth among a group of independently-living elderly persons in Japan, and to determine its association with general disease, medication, and dental status, as well as its effect on oral function. The study population consisted of participants of the Senior Citizens' College. The subjective sensations of oral dryness on waking and while eating a meal were measured by a questionnaire. The number of usable questionnaires was 1003 or 77.9%. The mean age of the subjects was 66.3 +/- 4.2 years, and 53.0% were male. More than one-third (37.8%) of the subjects reported oral dryness on waking. Only 9.1% of them noticed a subjective feeling of dry mouth during eating. Persons who had at least one of these symptoms made up 41.0%. A multiple stepwise logistic regression analysis indicated the following results: Perception of dry mouth on waking was more frequent among males (p < 0.001), persons who had a low BMI (p < 0.05), and those taking two or more prescribed drugs (p < 0.01). Sensation of dry mouth when eating was more frequent among subjects with a low BMI (p < 0.001) and those who wore a denture in the maxillary arch (p < 0.05). Perception of dry mouth when eating was associated with self-assessed chewing ability (p < 0.01) and dissatisfaction with speaking clearly (p < 0.05), as well as dental status. However, dissatisfaction with tasting a meal had a significant relationship with the reports of mouth dryness on waking (p < 0.01). Our findings suggest that a substantially higher percentage of persons have the perception of dry mouth on waking than when eating, which was associated with medications, being male, and having a low BMI. This perception may influence oral function, especially the reported dissatisfaction with tasting foods. PMID- 11484582 TI - Amlodipine-induced gingival overgrowth: periodontal responses to stopping and restarting the drug. AB - A case history of a woman with gingival overgrowth (GO) induced by amlodipine is presented. A 49-year-old Japanese woman, who was taking amlodipine, had gingival overgrowth and swelling on examination. No specific periodontal treatment was provided to the patient for the GO; however, the amlodipine was replaced with an ACE inhibitor after consultation with her medical practitioner. Within two months, the suspension of amlodipine resulted in a significant improvement in her periodontal condition. Failure to control the hypertension caused the physician to re-prescribe amlodipine. After three months, the gingival overgrowth returned; however, its severity was less when compared with the original periodontal condition, due to reduction in drug dose and periodontal therapy. This experience suggests that temporary suspension of a drug which can induce GO can improve the periodontal condition without the aid of surgical treatment. PMID- 11484583 TI - The index of ADOH: concept of measuring oral self-care functioning in the elderly. AB - Measures of ability, or function, are used in the health field to determine the level of performance of a specific task, assess the loss of function, and to measure the return to function as a result of rehabilitation or interventions. While these measures are important to all age groups, it is the elderly and chronically ill who are at most risk for adverse changes in their health and functional status. The Index of the Activities of Daily Oral Hygiene (ADOH) is an instrument to assess and monitor functional dependency in manipulating aids used in oral self-care. It is part of the overall concept of oral self-care which subsumes function, education and training, and compliance in achieving and maintaining oral health. The purpose of the Index of ADOH is two-fold: to assess a subject for the progressive loss of the physical ability to manipulate the aids used in oral self-care, and to measure the return to function in response to care intervention and rehabilitative services. These manipulative tasks are associated with flossing, brushing, topical fluoride application, and the use of oral rinses. Aside from changes in function, the Index of ADOH can be used by health care providers and administrators to project the need for services or support for home care. PMID- 11484585 TI - A method of eliminating streak artifacts from metallic dental restorations in CTs of head and neck cancer patients. AB - The purpose of this report was to develop a method to reduce streak artifacts derived from dental restorations in CT-imaging of the head and neck region. We selected six patients who were being treated for head and neck cancer and who had metal-derived artifacts on their CT that limited pre- or post-surgical visualization. The metal restorations were removed and replaced with non-metallic composite resin restorations. The CT was repeated as required on completion of the procedures. The streak artifacts were completely eliminated in patients in whom radiolucent composite materials were used. Patients have been followed for 6 26 months with no complications or breakdown of the restorations. The substitution of radiolucent for metallic restorations in special cases should be considered as a solution to CT dental artifact problems. PMID- 11484584 TI - The Oral Care Link Nurse: a facilitator and educator for maintaining oral health for patients at the Royal Hospital for neuro-disability. AB - This paper describes an ongoing oral health education project developed and implemented at the Royal Hospital for Neuro-disability (RHN) by the Dental Team. The Oral Care Link Nurse (OCLN) project was developed to assist the dental team in the promotion of oral care within the hospital. The Dental Team identified that, in addition to providing knowledge and skills to perform effective oral care, it is important that nursing staff retain positive attitudes and behavior toward providing that care. In conjunction with the Quality Assurance department, oral care guidelines were introduced throughout the hospital, with further input being focused on the training of at least one unqualified nursing auxiliary from each ward. These became the OCLN who were trained by the Dental Team to assist with the promotion of good oral care practices on the wards and to aid communication regarding patient oral health within the multidisciplinary team. Formal evaluation, by clinical audit, indicated that the project continued to improve the overall standard of oral care throughout the hospital. To ensure that this high standard continues, it is essential that continued support, commitment, and enthusiasm be given from the dental team and that support is constant. PMID- 11484586 TI - Prevalence of numeric anomalies in the permanent dentition of patients with Down syndrome. AB - The purpose of this study was to evaluate the prevalence of numeric anomalies in the permanent dentition of patients with Down syndrome by means of panoramic radiographs. The sample consisted of panoramic radiographs from 70 subjects. We examined the radiographs to detect hypodontia as well as supernumerary teeth. Our results confirm the high prevalence of hypodontia among patients with Down syndrome (60%), mostly with mild expression. The teeth most often missing were the upper lateral incisors, usually bilaterally, followed by the lower second premolars and upper second premolars. Supernumerary teeth were seen in 6% of the subjects, and the concomitant occurrence of hypodontia and supernumerary teeth occurred in one patient. PMID- 11484587 TI - You've gotta be kidding. PMID- 11484588 TI - Try family planning. PMID- 11484590 TI - National Practitioner Data Bank reporting requirements. PMID- 11484589 TI - Who's a rat? PMID- 11484592 TI - Your new practice resource: smilemichigan.com. PMID- 11484591 TI - New MDA president Dr. Bill Chase: "my mission is member value". Interview by Michael Maihofer. PMID- 11484593 TI - Oral pathology quiz. Case 1. Necrotizing sialometaplasia. PMID- 11484594 TI - Oral pathology quiz. Case 2. Florid cemento-osseous dysplasia. PMID- 11484595 TI - Oral pathology quiz. Case 3. Periapical cemento-osseous dysplasia--no treatment recommended. PMID- 11484596 TI - Oral pathology quiz. Case 4. Squamous cell carcinoma. PMID- 11484597 TI - Don't rely too much on data bank as major source for credentialing info. PMID- 11484598 TI - Project targets medical waste left on food trays. PMID- 11484599 TI - Patients should receive pain management info. PMID- 11484600 TI - Plan and execute your projects more effectively. PMID- 11484601 TI - [Proceeding of the 1999 and 2000 Congresses of the French Dendritic Cell Club]. PMID- 11484602 TI - [Dendritic cells and the control of the immune response]. AB - Cells of the dendritic family are suited to perform two distinct functions at two discrete locations. In the peripheral tissues, DC act as sentinels for "dangerous" antigens. They then migrate into the lymphoid organ, where they initiate activation of T lymphocytes which are specific for these antigens. During their migration, DC shift from an antigen-capturing-mode to a T cell sensitizing mode. In addition to switching on the immune response, subtypes of DC appear to influence the character of T cell differentiation, i.e. the Th1/Th2 balance. PMID- 11484603 TI - [Langerin: a new lectin specific for Langerhans cells induces the formation of Birbeck granules]. AB - Generation of monoclonal antibodies restricted to human dendritic cells generated from CD34+ hematopoietic precursors has enabled the identification of Langerin, a Ca(++)-dependent type II lectin. Only expressed by Langerhans cells, Langerin is responsible for Birbeck granule formation by membrane superimposition and zippering. Furthermore, cell-surface Langerin is rapidly internalized into Birbeck granules, and does not colocalize with MHC class II rich compartments. Langerin gene transfected into mouse fibroblasts induces the formation of Birbeck granule-like structures, that would permit a better understanding of the function of Birbeck granules. PMID- 11484604 TI - [Human thymic dendritic cells]. AB - Human thymic dendritic cells (DC) were enriched from thymocyte suspension by low density fractionation and elimination of CD3, CD8, CD19, CD56 and CD34-positive cells. Flow cytometry analysis shows that they belong to two distinct populations. The prominent (2/3) one is CD4+ CD45RA+ CD33-CD11c-, mostly immature as it lacks CD80, CD83 and CD86, and is HLA-DRint. High expression of CD123 and expression of pre-TCR-alpha transcripts links this subset to lymphoid, interferon producing cells. The other DC are typically CD4+ CD33+ CD11c+ myeloid cells mostly mature CD83+ HLA-DRhi. Both subsets could be generated in vitro from thymic CD34+ progenitors. Finally although experimental evidence ascribed to thymic DC a major role in establishing central tolerance through deletion of self reactive thymocytes, the respective functions of human lymphoid and myeloid subsets in the human thymus remain to be established. PMID- 11484606 TI - Design of engineered vaccines for systemic and mucosal immunity to HIV. AB - To design vaccines for viruses such as HIV that do not elicit sufficient protective immunity, we first constructed cluster vaccines containing T helper, CTL and neutralizing antibody epitopes. For second generation vaccines, we increased responses by enhancing binding to Major Histocompatibility molecules or by incorporating cytokines. We found that high avidity CTL induce better viral clearance. We also induced anti-HIV mucosal T cell immunity by intrarectal administration. Such approaches may improve classic attenuated or killed pathogen vaccines. PMID- 11484605 TI - [Dendritic cells and hepatitis C virus]. AB - Hepatitis C virus (HCV) induces chronic persistent infection that can lead to the development of hepatocellular carcinoma. We have searched for the presence of HCV genomic RNA in cells from hematopoietic origin and have, among others, documented such sequences in B cells as well as dendritic cells (DC) derived from monocytes. The allostimulatory capacity of these latter cells was found altered in chronic patients while it appeared restored in long term responders to therapy. PMID- 11484607 TI - [Dendritic cells and interaction with other cell types. Immune tolerance]. AB - T cell tolerance to self antigen is mainly established in the thymus were self reactive T cells are deleted. Interdigitating dendritic cells and medulary epithelial cells are directly involved in the deletion process. Some self reactive T cells escape, however this thymic censorship and enter the peripheral pool of naive T cells. Multiple mechanisms are also at play in the periphery to control this potentially armfull T cells, this include deletion and immune deviation. PMID- 11484608 TI - [Role of dendritic cells in the modulation of innate effectors of immunity]. AB - CD8+ cytotoxic T lymphocytes (CTL), specifically directed against tumor associated antigens, can be used in immunotherapy as effector cells in order to induce antitumor immune response. However, natural killer (NK) cells, that belong to the innate immune system, might also play a role on the anti-tumoral immune response. Our data show that quiescent NK cells can be activated by direct cell contact with dendritic cells (CD). Such a NK cells activating ability places DC at the frontier between innate and cognate immunity and then may encourage their use in clinical trials designed to elicit both CTL and NK responses. PMID- 11484609 TI - Dendritic cells in models of tumor immunity. Role of Flt3 ligand. AB - Cancer immunotherapy might be based on the administration to cancer patients of dendritic cells (DC) 'pulsed' with tumor-specific antigens. An alternative approach is to directly expand and/or activate DC in vivo using the cytokine Flt3 Ligand (FL). In mice, FL can drive large expansion of both lymphoid-related DC that appears to selectively enhance Th1-like immune responses and myeloid-related DC that enhances a more mixed Th phenotype. Immunization of FL-treated mice with a protein antigen leads to increased production of antibodies specific for that protein as well as to antigen-specific helper T cell responses. Studies of mouse tumor models have demonstrated that FL administration leads to the generation of protective anti-tumor immune responses, these effects being mediated by CTL and/or NK cells. When, FL has only minor or short term effects, the anti-tumor response can be significantly amplified by adding other cytokines, known to act at the T cell level, such as CD40 ligand (CD40L) or 4-1BBL, a TNF family member. Thus, combination of FL + CD40L or FL + 4-1BBL have superior anti-tumor effects vs. either cytokine alone. In conclusion, cytokines offer a variety of novel approaches for the treatment of cancer, infectious or auto-immune diseases. PMID- 11484610 TI - [Proteases, natural protease inhibitors and activation of the machinery of antigen presentation in dendritic cells]. AB - Dendritic cells play a central role in the immune response due to their exceptionally strong capacity for presenting antigens to naive T lymphocytes. Recent evidences have demonstrated that dendritic cells (DCs) exhibit a remarkable pattern of differentiation (maturation) that is accompanied by striking changes in morphology, organization and function. The hallmark of DC maturation is the major reorganization of the MHC class II molecule intracellular transport which is in part regulated by endosomal proteases activation. The central role of the endosomal proteases in generating antigenic peptides and controlling MHC class II traffic clearly defines these enzymes as an important area of investigation. PMID- 11484611 TI - Dental unit waterlines: review and product evaluation. AB - Dental practitioners must be knowledgeable regarding microbial contamination and biofilm formation in dental unit waterlines. Education should stress the need for improvement in the quality of water delivered to patients during treatment. Manufacturers must also play an important role by providing training and education regarding the proper use and maintenance of their systems. Dental facilities, both public and private, need reliable methods to prevent the development of biofilms within DUWs. These methods must be economical and require minimal effort to use on the part of the dental staff. In order for the system to work efficiently, the effluent water that is produced must be compatible with dental materials and be potentially free from toxic or carcinogenic materials. There are numerous models of water filtration units and chemical flushes available to the dental practitioner. However, the Food and Drug Administration have not approved all products currently on the market. Our evaluation of Zerosil, a new waterline-cleaning product, indicates that it is very easy to use and is extremely effective in killing the commonly found microorganisms in dental unit waterlines, as well as eliminating existing biofilms. It is also economical and requires minimum staff time to keep the waterlines clean. Following the initial treatments during week one, the water emanating from the DUWs was free from any viable microorganisms. This effect was present the entire three weeks in which the waterlines were treated. The elimination of viable microorganisms continued into a fourth study week, even though no further treatment of the DUWs was performed. Although the manufacturer recommends weekly treatment of DUWs following the initial treatment regimen, this result indicates that the product has a longer lasting effect than previously thought. Finally, the product can be delivered through any of the commercially available reservoir/bottle water delivery systems. From our study, Zerosil appears to meet the demanding requirements of keeping dental unit waterlines clean. Based on the research that has been done thus far, no universal treatment protocol can be recommended. A combination of approaches may offer the best available assurance of high-quality dental treatment water. Independent water reservoir systems, when used with a periodic chemical treatment protocol, have demonstrated safety and efficacy. Until we reach a point when a recommendation based on thorough evaluations can be made, dental offices should follow current ADA, OSAP, and CDC guidelines: flush waterlines for two to three minutes at the beginning of each day and for 20 to 30 seconds between each patient, and anti-retraction valves should be installed to prevent oral fluids from being drawn into dental waterlines. It is expected that in the near future, the dental practitioner will have a choice of proven systems and products to deal with this issue. Until that time, one should carefully evaluate any product or system being considered to prevent the formation of biofilms in DUWs. PMID- 11484612 TI - Oral pathology quiz #32. Case 1. Malignant salivary gland neoplasm. PMID- 11484613 TI - Oral pathology quiz #32. Case 2. Benign salivary gland tumor. PMID- 11484614 TI - Oral pathology quiz #32. Case 3. Cemento-osseous dysplasia. PMID- 11484615 TI - Oral pathology quiz #32. Case 4. Radicular cyst. PMID- 11484616 TI - Healthcare providers and the public rate the issues. PMID- 11484617 TI - Shorter waiting times for hip and knee replacement on the horizon. PMID- 11484618 TI - Telehealth, technology and nursing practice: complementary futures. PMID- 11484619 TI - Is there a Canadian tragedy in the making? PMID- 11484620 TI - e-health consumer: a diminishing tolerance of hospitals. PMID- 11484621 TI - . . . the makings of a potent brew of clashing cultures. PMID- 11484622 TI - The changing dynamics of the patient-provider relationship. PMID- 11484623 TI - Development of the Resident Assessment Instrument--Mental Health (RAI-MH). PMID- 11484624 TI - Commentary: refinement required before implementation. PMID- 11484625 TI - Commentary: RAI-MH holds great future promise. PMID- 11484626 TI - Commentary: the promise of better care. PMID- 11484627 TI - The implications of the new federal privacy legislation for public hospitals. PMID- 11484628 TI - Chemical decontamination plan for an emergency department. PMID- 11484633 TI - IRS private letter ruling addresses effect of ancillary joint ventures on an entity's tax exempt status. PMID- 11484632 TI - The duty to disclose HMO physician incentives. PMID- 11484634 TI - Pharmaceutical industry pricing practices: impact of the Bayer settlement. PMID- 11484635 TI - Understanding and treating drug abuse and addiction. PMID- 11484636 TI - Targeting the other STD. PMID- 11484637 TI - The legal advantage of disability management. AB - Actively working with disabled workers to try to accommodate their needs and limitations--even if accommodation proves impossible--provides powerful protection from ADA lawsuits. PMID- 11484638 TI - Pharmacy benefit managers use the Internet for savings. PMID- 11484640 TI - Creatively managing continuous improvement. PMID- 11484639 TI - Motorola's HAP-iness. PMID- 11484641 TI - Junior doctors' views on clinical audit--has anything changed? AB - A postal questionnaire survey of junior doctors' views was conducted in a large acute hospital in the south-east of England, amongst 146 junior medical staff recorded as being employed by the Trust across 21 specialities. It profiled their level of participation in audit and the quality of current audit programmes within their specialities and assessed their knowledge and understanding of clinical governance. Our findings suggest: a high level of involvement in activities labelled audit, but that these activities did not necessarily conform to robust audit methodologies; that junior doctors' professional attitudes towards clinical audit are influenced by negative experience of undertaking audit within their specialities; and that there was a variety of understanding about the principles and meaning of clinical governance. It concluded that the conditions for coherent strategy aimed at promoting effective audit programmes which could support the use of clinical audit as a tool for continuous professional development are not yet in place across the Trust. PMID- 11484642 TI - Quality management in German health care--the EFQM Excellence Model. AB - A brief insight into the issues affecting German health care is given. Namely, upward pressure on costs leads to increasing expenditure within health care and downward pressure from economic recession and political unwillingness to increase taxes and budget spending. In the case of competing key stakeholder perspectives, a quality management framework is seen as a potential approach for addressing these issues. The framework is the European Foundation for Quality Management (EFQM) Excellence Model. The article briefly explains the pilot phase for utilising the EFQM Excellence Model within German health care organisations and the various implementation approaches recommended by EFQM. Specific details of an approach used by a health care organisation within Germany (providing social medicine, geriatric and elderly services) are described in detail. The benefits achieved from using the Model are a clear picture of where the gaps are in the organisation and a clear vision for concentrating future efforts towards total quality management. PMID- 11484643 TI - Addicted to quality--winning the Dutch Quality Award based on the EFQM Model. AB - The Jellinek Center is a treatment service for prevention, care and cure of clients with addiction problems. Ten years ago the first quality improvement program was started. In 1994 the management team did the first formal self assessment based on the European Foundation for Quality Management (EFQM) Model and three assessors from the "Instituut Nederlandse Kwaliteit" audited the Jellinek and delivered a feedback report which was used to make an elaborate two year work-plan. Five projects to improve the quality of the Jellinek Center were identified and carried out. After two years, the work-plan was evaluated and the improvements assessed through a self-assessment conducted by the management team and an external audit team made up of six assessors. The findings of the self assessment were presented to the Dutch jury for the EFQM Award and the Jellinek Center won the Dutch Quality Award. The Jellinek is not complacent, however, as it recognises that the next phase of the quality management journey is to carry out self-assessment in all teams, redesign the treatment processes and improve performance measurements. PMID- 11484644 TI - Quality system based on the standard SFS-EN ISO 9002 in Kuopio University Hospital. AB - Kuopio University Hospital, situated within middle-east Finland, adopted the ISO 9002 standard as its quality system and gained formal certification in March 1999. The rationale behind the decision to adopt ISO 9002 is given, along with the main elements of the journey. The experiences of the hospital, including the advantages and disadvantages, are explained. In particular, issues regarding the documentation process, control and calibration of 4,000 pieces of medical equipment and the impact on staffing levels for in-house trained personnel to undertake the audits are described. The impact on the service to date, including benefits and drawbacks, is covered, along with aspirations for the future. Kuopio Hospitals adopted some techniques during the implementation process which did not work as successfully as others. The article therefore includes these in an effort to pass on the learning acquired from implementing ISO 9002 within such a large hospital as Kuopio University Hospital. PMID- 11484645 TI - Comparing patients' experience of mental health services in England: a five-Trust survey. AB - The implementation of the Care Programme Approach (CPA) in English mental health services has been slow to proceed despite general support, both in England and in other countries, of its principles of good practice. This study set out to evaluate the implementation of the CPA directly from patients' experience using the "Your Treatment and Care" assessment tool. The results of a survey of 503 patients across five NHS Trusts in England showed that many patients did not have a copy of their care plan and had not been involved in the care planning procedure. Many reported shortcomings in their experience of their key worker and their psychiatrist. However, there was substantial variation in experience across services. "Your Treatment and Care" showed good internal reliability, was acceptable to users, and appeared to be able to access actual experiences better than a traditional "satisfaction" item. It appears to be very useful as a benchmarking tool and is now being used in services across the UK, the USA and Australia. PMID- 11484646 TI - Quality and the appraisal process. PMID- 11484647 TI - A study of patients' expectations and satisfaction in Singapore hospitals. AB - In today's highly competitive healthcare environment, hospitals increasingly realise the need to focus on service quality as a means to improve their competitive position. Customer-based determinants and perceptions of service quality therefore play an important role when choosing a hospital. This paper attempts to determine the expectations and perceptions of patients through the use of a generic, internationally used market research technique called SERVQUAL. An analysis covering 252 patients revealed that there was an overall service quality gap between patients' expectations and perceptions. Thus, improvements are required across all the six dimensions, namely, tangibility, reliability, responsiveness, assurance, empathy and accessibility and affordability. PMID- 11484648 TI - The assessment of quality in medical diagnostic tests: a comparison of ROC/Youden and Taguchi methods. AB - Diagnostic tests are widely used in many areas of modern technological society, but they are of particular importance in medicine, where early and accurate diagnosis can decrease morbidity and mortality rates of disease. How the quality of diagnostic information and decisions should be measured in a meaningful way has become increasingly important in recent years as an abundance of new diagnostic tests have been introduced. A number of seemingly independent indices are studied for evaluating diagnostic performance such as the receiver operating characteristic curves and signal-to-noise ratios. Designing robustness into diagnostic tests can only be achieved by minimizing the variation in the total number of false diagnosis. This article has undertaken a comparison of signal-to noise ratios developed by Taguchi in quality engineering and system performance in manufacturing industry. A hybrid is also computed and its relevance to physicians as an efficient assessment method is proposed and strongly encouraged. PMID- 11484649 TI - Responding to diversity: the Primary Care Group in the NHS. AB - The paper attempts to examine the development of Primary Care Groups in the NHS, utilising a conceptual framework taken from relationship marketing. In particular, it looks at a framework representing the complex relationships between the Primary Care Group and a diverse range of internal and external stakeholders and the implications of these relationships. It reports the preliminary findings of an ongoing, in-depth case study of two Primary Care Groups; with data collected from in-depth interviews with a small number of key stakeholders. The results, so far, suggest the importance of developing and maintaining longer-term relationships with a range of partners, both internal and external. It recommends that Primary Care Groups should develop strategies to work closely with these stakeholders, as an essential underpinning to developing continuous improvement in performance, quality and "user" retention. PMID- 11484650 TI - The theory and methodology of provider profiling. AB - Physician profiling is the comparison of physician practice patterns to determine the existence and effects of significant differences in outcomes. To optimize care quality, these comparisons can be used to influence provider behavior through awareness when outcomes are poor or do not warrant provider expenditures. To maximize the value of such a technique, the underlying bases for comparison and the uses of derived data must be understood. Several factors must be considered or controlled for when determining comparable providers. Additionally, the worth or significance of findings must also be understood. This paper seeks to provide information in these two areas for the benefit of those responsible for managing care at all levels. PMID- 11484651 TI - [Food habits and oral hygiene of students. Investigation of 150 students in university housing]. AB - The objective of this study is to describe the food intake and the oral hygiene habits of 150 Senegalese students living in a university campus. According to Miller, we realised a four days survey. We noted an increase of sugar drinks; tea is the most consumed drink. The results showed also that the toothbrush as means of oral hygiene supplanted the traditional means such as "cure dents" or water rinse with finger. PMID- 11484652 TI - [Oral hygiene habits and dental caries among students. Investigation of 150 students in university housing in Dakar, Senegal]. AB - The objective of this study is to describe the oral hygiene habits and the status of the teeth decay of 150 senegalese students living in a university campus. The results showed that the toothbrush as mean of oral hygiene supplanted the traditional means such as "cure dents" or water rinse with finger. Results showed also a high rate decay (60%) but the CAO index remained low (2.04%). We founded an improvement of dental decay status with an increasing daily number of tooth brushing: we noted a decreasing dental plaque deposition rate, a decreasing decay rate and a CAO index remaining low until adult age. PMID- 11484653 TI - Third molar impaction: evaluation of the symptoms and pattern of impaction of mandibular third molar teeth in Nigerians. AB - Three hundred and thirty eight patients between the ages of 16 and 54 years (mean 24.4, standard deviation 6.1 years) were seen. They presented with 473 impacted mandibular third molars. A total of 341 (72.09%) impaction were seen in patients between the ages of 16 to 25 years, while 420 (88.8%) impaction were seen between the ages of 16 to 30 years. Assessing the level of impaction using Pell and Gregory classification showed that 358 (54.55%) impaction were in position A, 151 (31.92%) were in position B while 64 (13.53%) were in position C. One hundred and seven (22.62%) were in position I, 288 (60.89%) were in position II, while 78 (16.49%) were in position III. Of the 473 impacted mandibular third molars, 323 (68.29%) had symptoms of pain. Two hundred and seventy two (57.51%) impacted teeth were associated with pathology. Out of these, 203 (42.92%) were pericoronitis and periodontal disease, 66 (13.95%) were caries, while 3 (0.63%) were associated with cysts. In conclusion, the level of impaction suggests that a remarkable number of impacted mandibular third molars should be removed under general anesthesia. There is need for further studies to determine the levels of impaction, the types of anesthesia used during extraction and the outcome. PMID- 11484654 TI - Evaluation of precautions adopted by dental surgeon using local anaesthesia. AB - There were 62 surgeons consisting of 38 (61.3%) males and 24 (38.7%) were females. Thirty-one (50%) were from private and state government owned clinics while the rest were from teaching hospitals. Thirty-three (53.2%) were dental house surgeons, senior dental house surgeons and dental officers, while 29 (46.8%) were within the rank of registrar and senior dental officer, senior registrar, principal dental officer and consultant. Six (9.7%) use aspirating syringe all the time. 11 (17.7%) use aspirating syringe occasionally and 45 (72.6%) use non aspirating syringe. All the surgeons wear facemasks and latex gloves. On the replacement of the needle guard after injection, 58 (93.5%) indicated that the needle is first inserted into the needle guard and then secure 4 while 4 (6.5%) pick-up the guard with their fingers, place it over the needle and secure the guard. Eight (12.9%) indicated that the maximum dose of 2% lignocaine with adrenaline 1:80,000 is 7 mg/kg body weight or less, 5 (8.1%) indicated 10 mg/kg body weight, while 49 (79.0%) did not complete this section. On the maximum number of 1.8 ml cartridges, all the surgeons indicated that the maximum is 12 or fewer cartridges. This study revealed that the risk of intravascular injection is high. Although of the most dental surgeons take necessary precautions to avoid complications arising from the use of local anaesthetics, there is a need for total compliance in view of fatal complications that may ensue. It also underscores the need for continuous dental education program to update practitioners. PMID- 11484655 TI - [Outcome of endodontic retreatments at the Institute of Ondonto-Stomatology of Dakar (from 1986 to 1997]. AB - The retrospective study on the endodontic retreatments done at the Odonto Stomatology Institute of Dakar from 1986 to 1997 shows a small rate of 1.94% (unreached canalar root filling) for 4325 endodontic treatments. The endodontic retreatments concern almost women (54.76%), ageing between 20 to 30 years (45.24%); maxillary central incisors (21.43%) and first molars (21.43%). The clinical reason having conducted to a retreatment is prosthetic (46.4%) and symptomatic (38.10%); the immediate success rate of these retreatment stays favourable at 59.52%. PMID- 11484656 TI - [Chronology and date of eruption of primary teeth in Senegal]. AB - Chronology and eruption dates of primary teeth are parameters of importance in different medical scientific fields. We carried a longitudinal clinical study to determine these parameters in a local Senegalese population. The study interests a sample of 573 Senegalese Negro children of the two sexes and its aims are to situate these parameters formally and in space in on hand, and to be able to justify; in comparison to these parameters, the disturbances which accompany often the phenomenon of dental eruption in the other hand. We have results relating to chronology and eruption date of the 20 primary teeth of each of the children. Our study strengthens data already known but it reveals also other data as well as important. Other studies are however necessary to set a good board of chronology and eruption date of primary teeth in the Senegalese population. PMID- 11484657 TI - [Clinical study of dental and periodontal infectious complications observed at the Central Hospital of Yaounde--apropos of 161 cases]. AB - The septic condition resulting from an apical infection, a peridontal infection, an accident of dental eruption or post-extraction can be disseminated and can lead to complications. A retrospective study of 161 cases admitted at the Central Hospital of Yaounde from 1995 to 1998 was done with the aim of establishing a distribution according to sex, age, profession, infectious stage and causes of late consultation as well as the relation between the infectious stage, aetiology, age and profession. It was observed that: the age group mostly affected were: 10-19 years (25.46%) and 20-29 years (30.43%), it was at the late stage of abscess (54.03%) and of phlegmon (32.91%) that the patients came for consultation and this was primarily due to poor socio-economic conditions (55.90%), abscess was formed mostly among the students of 10-19 years and 0-9 years while phlegmon was observed mostly among the students between 20-29 years. It was for these reasons that we suggest: a better prevention of infectious dental diseases in schools and universities, a sensibilisation of parents and educators on the economic interest of early treatment of these infections. PMID- 11484658 TI - Should we ever do sham operations? PMID- 11484659 TI - Outcome prediction in severe head injury: analyses of clinical prognostic factors. AB - Retrospective analysis of 272 patients with severe head injury was performed. Patient age, Glasgow Coma Scale (GCS) score, pupillary abnormalities, impaired oculocephalic response, presence of subarachnoid haemorrhage, and multiplicity of parenchymal lesions on computerised tomography (CT) were examined. The CT findings were divided into 2 groups, diffuse brain injury (DBI) and mass lesion, according to the classification of the Traumatic Coma Data Bank. The DBI, basically, has no high or mixed density lesion more than 25 ml on CT, and was classified into 4 subgroups: DBI I includes injuries where there is no visible pathology; DBI II includes all injuries in which the cisterns are present with a midline shift of less than 5 mm; DBI III includes injuries with swelling where the cisterns are compressed or absent and the midline shift is less than 5 mm; DBI IV includes injuries with a midline shift of more than 5 mm. The mass lesions were categorised into 3 subgroups: epidural haematoma; acute subdural haematoma; and intracerebral haematoma. Outcomes were determined at 6 months following trauma using the Glasgow Outcome Scale. All DBI I patients recovered well. In the DBI II group, age, GCS score and detection of multiple parenchymal lesions on CT were significantly correlated with outcome. For the DBI III and IV groups, the only significant prognostic factor was the GCS score. In patients with a mass lesion, the GCS score was the only significant prognostic factor in the epidural haematoma group, but the GCS score and the presence of subarachnoid haemorrhage were predictive factors in the acute subdural haematoma group. Outcomes were unfavourable in the majority of patients with intracerebral haematoma. GCS score could predict outcome in all groups. The confidence of the outcome prediction ranged from 75.8 to 92.1%, depending on logistic regression analysis. PMID- 11484660 TI - Outcome following anterior cervical discectomy in compensation patients. AB - This is a retrospective study aimed to analyse the clinical outcomes of patients following anterior cervical decompression and fusion for radiculopathy in worker's compensation, third party and non-compensable group. The outcome of 224 cases operated between 1991 to 1998 were analysed. Only patients with radiculopathy due to a cervical disc protrusion and spondylosis were included. There were 140 non-compensable patients, 58 worker's compensation and 26 third party. There was no statistical difference in radiological fusion between the three groups (P=0.46). The worker's compensation and third party claimant groups, had an 'excellent' outcome at 65% and 69% respectively, compared to the non compensation group at 79% (P=0.042). Rates of poor outcome were high in the worker's compensation group (9%) compared with third party (4%) and the non compensable group (5%). Financial incentives seem to significantly influence the outcome of cervical disc surgery in our patient population. PMID- 11484661 TI - Intracranial bleeding rates associated with two methods of external ventricular drainage. AB - We investigated the risk of intracranial haemorrhage with two frequently performed methods of external ventricular drainage (EVD). Haemorrhage is believed to be a rare complication of such procedures, although in most studies reported in the literature standardised evaluation of computed tomography (CT) scans was not performed in all cases. Data were analysed retrospectively for 82 patients who had undergone percutaneous needle trephination and 92 who had undergone classic ventriculostomy. We found an overall bleeding risk with EVD of 9.4%. Most haematomas were small, and only one caused neurological symptoms. Therefore, the risk of symptomatic haemorrhage in this study was low (0.5%). Percutaneous needle trephination was associated with a higher risk of haemorrhage (12.2%) than classic ventriculostomy (6.5%), although differences were not statistically significant. We conclude from our results that small asymptomatic haemorrhages occur far more often after EVD than has generally been suspected. To compare the true risk of bleeding associated with different drainage methods requires controlled studies in which CT scans are evaluated in a standardised way. PMID- 11484662 TI - Acute neurologic deterioration after surgical treatment for thoracic spinal stenosis. AB - Spondylotic changes in the thoracic spine resulting in a myelopathic syndrome is a rare condition. Surgical treatment consists of removal of the hypertrophied ligaments, facet joints and laminae (laminectomy). In this report, we review our experience in treating 12 cases of thoracic spinal stenosis (TSS). From 1989 1998, we identified 12 surgical cases treated by the Department of Neurosurgery at Temple University Hospital. For this report, we reviewed both inpatient and outpatient records and radiographic studies. We also reviewed the world's literature regarding this entity. After reviewing our own cases of TSS and the world's literature, we identified a higher than expected incidence of acute neurologic deterioration (AND) after surgical treatment of TSS (14.5%). This incidence is compared to an expected incidence of AND (0-5.5%) after laminectomy for cervical spinal stenosis (CSS). The seemingly higher incidence of AND after surgical treatment of TSS has implications for discussion of risks with patients undergoing surgical treatment. PMID- 11484663 TI - Cranio-spinal enterogenous cysts: clinico-radiological analysis in a series of ten cases. AB - Enterogenous cysts are rare congenital lesions resulting from dysgenesis during the third embryonic week at the time of notochord development and the transitory existence of the neurenteric canal. Enterogenous cysts are usually lined by mucus secreting epithelium resembling that of the gastrointestinal tract. The inclusion of the endodermal elements in the spinal canal may produce extradural, intradural extramedullary or intradural intramedullary cystic lesions. The endodermal inclusions producing cysts in the intracranial compartment are extremely rare, especially in the supratentorial region. Plain X-rays may show concomitant cranio spinal anomalies. However, CT and MRI scans show better definition and characterisation. Total excision is the procedure of choice in these benign cysts with favourable long term prognosis. Approximately 100 histologically proven cases have been described, with a few exceptions, as isolated case reports the authors report a detailed analysis of the clinico-radiological aspects of a series of ten patients with cranio-spinal enterogenous cysts with a review of the literature. PMID- 11484664 TI - Transcranial Doppler in carotid endarterectomy. AB - PURPOSE: To evaluate the role of transcranial Doppler monitoring in reducing the complications of carotid endarterectomy, by analysing: (1) perioperative intracerebral blood flow velocity; (2) embolic load; and (3) effect of Dextran-40 therapy on patient outcome. DESIGN: Retrospective case series. METHODS: The study cohort consisted of 30 consecutive patients undergoing 32 carotid endarterectomy procedures. Continuous transcranial Doppler (TCD) monitoring of the ipsilateral middle cerebral artery (MCA) was performed to obtain flow velocities and embolic count. Flow velocity changes were analysed in relation to electroencephalographic (EEG) changes. A Dextran-40 infusion was instituted for patients who experienced a post-arteriotomy embolic load of >50 counts/hour. RESULTS: The average middle cerebral artery velocity (MCAvel) drop on cross clamping was 46+/-12.1%, expressed as a percentage of the individual's 24-hour pre-operative value. Clamping ischaemia developed in six cases (18.8%) of which three (9.4%) demonstrated TCD changes only, and three demonstrated both TCD and EEG changes (9.4%). The average increase in MCAvel at 60-120 minutes postarteriotomy, was 18+/-17.5%; six cases developed hyperaemia. Postoperative emboli were seen in 88% of cases with 31% of patients demonstrating embolic loads of >50/hour. CONCLUSIONS: Perioperative transcranial Doppler monitoring (1) is a useful adjunct to EEG assessment of cross clamping ischaemia, providing information in real time, on MCA blood velocity deterioration that may antecede irreversible change (2) detects postoperative hyperaemic response, and (3) allows quantification of microembolic loads that, when high, have been shown to be a precursor to localised cerebral ischaemia and can be effectively treated with Dextran-40 infusion. PMID- 11484665 TI - Is the asterion a reliable landmark for the lateral approach to posterior fossa? AB - An anatomical study was conducted to gain orientation regarding the posterolateral approaches. The asterion is defined as the junction of the lambdoid, parietomastoid, and occipitomastoid sutures. This anatomical point has been widely used as a landmark in lateral approaches to posterior fossa. Although there are many common practices in posterolateral approaches, studies providing accurate anatomical knowledge as to what is the correct point to start a craniotomy are limited in number. Therefore, this study was conducted in an attempt to determine the reliability of the asterion for the posterolateral approaches as surgical landmark. PMID- 11484666 TI - Medulloblastoma/primitive neuroectodermal tumour studied as a Matrigel enhanced subcutaneous xenograft model. AB - An important role for pre-clinical models of medulloblastoma/primitive neuroectodermal tumour (MB/PNET) is inhibited by the limitations of conventional heterotransplantation. Nine cohorts of MB/PNET were studied for subcutaneous engraftment in nude mice by both conventional and Matrigel supplemented methods. While no subcutaneous tumours resulted from 63 conventional attempts, an aggregate 41 xenografts from 72 injections (57%) were produced when Matrigel was added to the cell suspension. In subsequent passage, engraftment rate approached 100%. To study the response to chemotherapeutic agents in the model, a total of 221 tumours in 3 cohorts were treated using one of the following: cisplatin, carboplatin, vincristine, cyclophosphamide, diaziquone, or saline control. While all agents demonstrated statistically significant activity, cyclophosphamide proved to be particularly effective. The potential applications of this xenograft model in the biologic as well as therapeutic study of MB/PNET deserve continuing investigation. PMID- 11484667 TI - The effect of magnitudes and duration of pressure on cerebral cortex in a rat model. AB - The aim of this biomechanical study was to investigate the pressure-time-damage relationship on the cerebral cortex using a rat model. During brain surgery, additional cortical injuries induced by traction have been an important clinical issue. A total of 84 rats underwent a unilateral craniectomy, and conduction by hydrostatic pressure loading through a modified central venous pressure device with various combinations of magnitudes ranging from 0.5 to 10 cm H2O and duration from 0.5 to 20 minutes was then performed. Histopathological examination has exhibited three patterns of clustered, spotted damaged neurons and undamaged neuron. Two best-fitted exponential curves were derived from the data to establish the damaged, critical and tolerable loadings responsible for the neuron viability. This research provides information to enhance understanding of the additional cortex injuries induced by traction. Furthermore, the results may have implications in providing clinical guidance and/or development of alarm systems for minimising cortical damage during surgery. PMID- 11484668 TI - An unprovoked tonic-clonic seizure should generally be treated. PMID- 11484669 TI - Abstracts of the 6th Meeting of the Italian Peripheral Nerve Study Group. Sondrio, 5-7 April 2001. PMID- 11484670 TI - 23rd Annual Carrell-Krusen Symposium. Dallas, Texas, February 22-23, 2001. Abstracts. PMID- 11484673 TI - Abstracts of the International Diabetes Federation Congress, 5th Diabetic Neuropathy Satellite Symposium. October 31-November 4, 2000. San Jose, Costa Rica. PMID- 11484674 TI - Brain Edema XI. Proceedings of the 11th International Symposium. Newcastle-upon Tyne, United Kingdom. June 6-10, 1999. PMID- 11484675 TI - Proceedings of The Biological Foundations of Music Conference. May 20-22, 2000. New York, New York, USA. PMID- 11484676 TI - Neuro-cardiovascular regulation: from molecules to man. Proceedings of the Baroreceptor and Cardiopulmonary Receptor Reflexes Conference. August 23-27, 2000. Iowa City, Iowa, USA. PMID- 11484677 TI - Antoine Bechamp: pere de la biologie. Oui ou non? AB - There is an alternative medicine lobby that, in conjunction with antivivisectionists, believes Louis Pasteur to have been a fraud [R. Bottomley's You Don't Have to Feel Unwell! (Newleaf, 1994) is a recent example]. They frame their accusations around a rivalry between Pasteur and a contemporary, Antoine Bechamp, from whom they suggest Pasteur stole his ideas and then distorted them for his own purposes. This article explores some aspects of the controversies between Bechamp and Pasteur. PMID- 11484678 TI - Peer led programme for asthma education in adolescents. Papers describing cluster randomised trials must be peer reviewed by statisticians. PMID- 11484679 TI - Peer led programme for asthma education in adolescents. Issues of design and analysis are crucial in cluster randomised trials. PMID- 11484680 TI - Alcohol and drug services have been patient centred for years. PMID- 11484681 TI - Minimal refereeing will lead to global equity of information. PMID- 11484682 TI - Study should have reported more data about associated diseases. PMID- 11484684 TI - Proceedings of the 43rd Annual Thomas L. Petty Aspen Lung Conference: Mechanisms of Pulmonary Fibrosis. PMID- 11484683 TI - Epidemiological surveillance of rubella must continue. PMID- 11484685 TI - Look before you leap. PMID- 11484686 TI - Efficiency after altitude acclimatization. PMID- 11484687 TI - A randomized, controlled trial of surgery for temporal-lobe epilepsy. AB - BACKGROUND: Randomized trials of surgery for epilepsy have not been conducted, because of the difficulties involved in designing and implementing feasible studies. The lack of data supporting the therapeutic usefulness of surgery precludes making strong recommendations for patients with epilepsy. We conducted a randomized, controlled trial to assess the efficacy and safety of surgery for temporal-lobe epilepsy. METHODS: Eighty patients with temporal-lobe epilepsy were randomly assigned to surgery (40 patients) or treatment with antiepileptic drugs for one year (40 patients). Optimal medical therapy and primary outcomes were assessed by epileptologists who were unaware of the patients' treatment assignments. The primary outcome was freedom from seizures that impair awareness of self and surroundings. Secondary outcomes were the frequency and severity of seizures, the quality of life, disability, and death. RESULTS: At one year, the cumulative proportion of patients who were free of seizures impairing awareness was 58 percent in the surgical group and 8 percent in the medical group (P<0.001). The patients in the surgical group had fewer seizures impairing awareness and a significantly better quality of life (P<0.001 for both comparisons) than the patients in the medical group. Four patients (10 percent) had adverse effects of surgery. One patient in the medical group died. CONCLUSIONS: In temporal-lobe epilepsy, surgery is superior to prolonged medical therapy. Randomized trials of surgery for epilepsy are feasible and appear to yield precise estimates of treatment effects. PMID- 11484688 TI - Mutation in the gene for bone morphogenetic protein receptor II as a cause of primary pulmonary hypertension in a large kindred. AB - BACKGROUND: Most patients with primary pulmonary hypertension are thought to have sporadic, not inherited, disease. Because clinical disease develops in only 10 to 20 percent of persons carrying the gene for familial primary pulmonary hypertension, we hypothesized that many patients with apparently sporadic primary pulmonary hypertension may actually have familial primary pulmonary hypertension. METHODS: In a study conducted over 20 years, we developed a registry of 67 families affected by familial primary pulmonary hypertension. Through patient referrals, extensive family histories, and correlation of family pedigrees, we discovered shared ancestry among five subfamilies. We established the diagnosis of primary pulmonary hypertension by direct evaluation of patients and review of autopsy material and medical records. We assessed some family members for mutations in the gene encoding bone morphogenetic protein receptor II (BMPR2), which has recently been found to cause familial primary pulmonary hypertension. RESULTS: We linked five separately identified subfamilies that included 394 known members spanning seven generations, which were traced back to a founding couple in the mid-1800s. Familial primary pulmonary hypertension has been diagnosed in 18 family members, 12 of whom were first thought to have sporadic disease. The conditions of 7 of the 18 were initially misdiagnosed as other cardiopulmonary diseases. Six members affected with familial primary pulmonary hypertension and 6 of 10 at risk for carriage have been undergone genotype analysis, and they have the same mutation in BMPR2, a transversion of thymine to guanine at position 354 in exon 3. CONCLUSIONS: Many cases of apparently sporadic primary pulmonary hypertension may be familial. Failure to detect familial primary pulmonary hypertension results from incomplete expression within families, skipped generations, and incomplete family pedigrees. The recent discovery of mutations in BMPR2 should make it possible to identify those with susceptibility to disease. PMID- 11484689 TI - Clinical and molecular genetic features of pulmonary hypertension in patients with hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Most patients with familial primary pulmonary hypertension have defects in the gene for bone morphogenetic protein receptor II (BMPR2), a member of the transforming growth factor beta (TGF-beta) superfamily of receptors. Because patients with hereditary hemorrhagic telangiectasia may have lung disease that is indistinguishable from primary pulmonary hypertension, we investigated the genetic basis of lung disease in these patients. METHODS: We evaluated members of five kindreds plus one individual patient with hereditary hemorrhagic telangiectasia and identified 10 cases of pulmonary hypertension. In the two largest families, we used microsatellite markers to test for linkage to genes encoding TGF-beta-receptor proteins, including endoglin and activin-receptor-like kinase 1 (ALK1), and BMPR2. In subjects with hereditary hemorrhagic telangiectasia and pulmonary hypertension, we also scanned ALK1 and BMPR2 for mutations. RESULTS: We identified suggestive linkage of pulmonary hypertension with hereditary hemorrhagic telangiectasia on chromosome 12q13, a region that includes ALK1. We identified amino acid changes in activin-receptor-like kinase 1 that were inherited in subjects who had a disorder with clinical and histologic features indistinguishable from those of primary pulmonary hypertension. Immunohistochemical analysis in four subjects and one control showed pulmonary vascular endothelial expression of activin-receptor-like kinase 1 in normal and diseased pulmonary arteries. CONCLUSIONS: Pulmonary hypertension in association with hereditary hemorrhagic telangiectasia can involve mutations in ALK1. These mutations are associated with diverse effects, including the vascular dilatation characteristic of hereditary hemorrhagic telangiectasia and the occlusion of small pulmonary arteries that is typical of primary pulmonary hypertension. PMID- 11484690 TI - The pseudomonas hot-foot syndrome. AB - BACKGROUND: Between March and May 1998, there was an outbreak of a clinically distinct skin eruption on the soles of the feet of children who used a community wading pool. METHODS: We reviewed the medical records of 40 children in whom this syndrome developed between March and May 1998. We treated 17 children and advised the attending physicians on the care of the other 23. Follow-up data were obtained for up to one year. RESULTS: Exquisitely painful erythematous plantar nodules developed in 40 children (age, 2 to 15 years) within 40 hours after they had used a wading pool whose floor was coated with abrasive grit. Culture of the plantar pustules from one child yielded Pseudomonas aeruginosa with a pattern on pulsed-field gel electrophoresis that was identical to that of a strain of P. aeruginosa cultured from the pool water. A skin-biopsy specimen from this patient showed a perivascular and perieccrine neutrophilic infiltrate, and a specimen from another patient showed a dermal microabscess. Thirty-seven patients were treated symptomatically; three others were treated with cephalexin. All patients recovered within 14 days, but three children had recurrences of the painful plantar nodules within 24 hours after using the pool again. Folliculitis developed in one patient. CONCLUSIONS: The "pseudomonas hot-foot syndrome" is characterized by the acute onset in children of exquisitely tender plantar nodules and a benign, self-limited course. This community outbreak developed after exposure to pool water containing high concentrations of P. aeruginosa. PMID- 11484691 TI - Images in clinical medicine. Nail injury to the skull. PMID- 11484692 TI - Autoimmune diseases. PMID- 11484693 TI - Cardiovascular complications of cocaine use. PMID- 11484694 TI - Clinical problem-solving. Of nicks and time. PMID- 11484695 TI - Finally, a randomized, controlled trial of epilepsy surgery. PMID- 11484696 TI - Genetic clues to the cause of primary pulmonary hypertension. PMID- 11484697 TI - Sacred secrets--the privacy of medical records. PMID- 11484698 TI - Letters of condolence. PMID- 11484699 TI - Letters of condolence. PMID- 11484700 TI - Letters of condolence. PMID- 11484701 TI - Letters of condolence. PMID- 11484702 TI - Letters of condolence. PMID- 11484703 TI - Letters of condolence. PMID- 11484704 TI - Paroxetine for the prevention of depression induced by interferon alfa. PMID- 11484705 TI - Human papillomavirus infection as a risk factor for squamous-cell carcinoma of the head and neck. PMID- 11484706 TI - Human papillomavirus infection as a risk factor for squamous-cell carcinoma of the head and neck. PMID- 11484707 TI - Human papillomavirus infection as a risk factor for squamous-cell carcinoma of the head and neck. PMID- 11484708 TI - Hematopoietic-cell transplantation for chronic granulomatous disease. PMID- 11484709 TI - Sepsis in a newborn due to Pseudomonas aeruginosa from a contaminated tub bath. PMID- 11484710 TI - Fulminant coxsackieviral myocarditis. PMID- 11484711 TI - The limits of state laws to protect genetic information. PMID- 11484712 TI - Images in clinical medicine. A hair-on-end skull. PMID- 11484713 TI - [Prevention of cardiovascular diseases:a medical-economic approach]. PMID- 11484714 TI - The role of digital fundus photography in diabetic retinopathy screening. Digital Diabetic Screening Group (DDSG). AB - PURPOSE: The role of digital monochromatic nonmydriatic fundus photography as an adjunct in the diagnosis of diabetic retinopathy is evaluated. METHODS: 197 patients were sequentially evaluated by three different techniques: dilated ophthalmoscopy by an experienced ophthalmologist, performance of 7 standard color mydriatic stereo fields, and a single digital monochromatic nonmydriatic image incorporating the disc and macula. Stereo color photographs served as the reference standard and were compared to either ophthalmoscopy performed by a physician, or a single digital photograph transmitted electronically to a reading site and evaluated by a trained non-physician grader. Sensitivity and specificity of the three methods were compared. The decision as to whether or not to refer to an ophthalmologist for potential treatment (Kaiser modified ETDRS level > 21) was then chosen for analysis. RESULTS: A single nonmydriatic monochromatic digital photograph appeared equivalent to standard color photography and more sensitive than mydriatic ophthalmoscopy in the detection of diabetic retinopathy in this patient population. Sensitivity of digital photography compared with color photography was 78%, and the specificity 86% contrasted with comparable ratios of 34% and 100% for ophthalmoscopy versus color photography. No patient identified by ophthalmoscopy alone for referral based on retinopathy level of > 21 would have been missed by a single digital monochromatic photographic image. CONCLUSION: A single nonmydriatic monochromatic wide field digital photograph of the disc and macula in diabetic patients is a sensitive and cost-effective means for detecting diabetic retinopathy in high-risk populations. PMID- 11484715 TI - [Diabetes, arterial hypertension and cerebrovascular accident]. PMID- 11484716 TI - Methods of applying manipulative technic. 1945. PMID- 11484717 TI - Soft tissues in areas of osteopathic lesion. 1947. PMID- 11484718 TI - The application of the respiratory principle to osteopathic manipulative procedures. 1949. PMID- 11484719 TI - New labeling and use changes for Rezulin. PMID- 11484720 TI - Measles, mumps, and rubella (MMR) vaccine and autism. MMR cannot be exonerated without explaining increased incidence of autism. PMID- 11484721 TI - Measles, mumps, and rubella (MMR) vaccine and autism. Argument is too simplistic. PMID- 11484722 TI - Using clinical evidence. Obstetricians seem to be overstating the evidence in major placenta praevia. PMID- 11484723 TI - Using clinical evidence. Clinical evidence is one of respondents' three favourite sources of information. PMID- 11484724 TI - Questionnaires for depression and anxiety. Routine screening entails additional pitfalls. PMID- 11484725 TI - NHS staff must be trained in how to prevent aggression. PMID- 11484726 TI - Questionnaires for depression and anxiety. Two screening questions may be helpful. PMID- 11484727 TI - Exposure of healthcare workers to bloodborne viruses. Eye and face protection should be used. PMID- 11484728 TI - [Foundations of the Monte Carlo method for dose calculation in radiotherapy]. AB - Monte Carlo (MC) methods applied in dose calculation are based on fundamental principles of radiation interaction with matter. In contrast to other methods, the accuracy of dose calculation achievable with MC depends only on the determination of the beam quality and the interaction coefficients. Using MC techniques it is possible to predict the dose for clinical photon and electron beams with an accuracy of > +/- 2%. Especially for inhomogeneous regions like head, neck, and lung, the MC technique can significantly improve the accuracy compared to conventional algorithms. Therefore, in the present paper the basic features of the MC method are reviewed in the context of treatment planning in radiation therapy. The main shortcoming in the past, i.e., that MC algorithms are too slow to be acceptable for clinical purposes, could be solved by using faster computers and by introducing new variance reduction (VR) techniques. These techniques decrease the statistical fluctuations without increasing the number of particle histories. Therefore, MC calculation times in the order of a few minutes are possible. A brief overview of VR methods is provided. PMID- 11484729 TI - From the Centers for Disease Control and Prevention. Evaluation of sexually transmitted disease control practices for male patients with urethritis at a large group practice affiliated with a managed care organization--Massachusetts, 1995-1997. PMID- 11484730 TI - From the Centers for Disease Control and Prevention. Measles incidence before and after supplementary vaccination activities--Lusaka, Zambia, 1996-2000. PMID- 11484731 TI - From the Centers for Disease Control and Prevention. Influenza and pneumococcal vaccination levels among persons aged >/= 65 years--United States, 1999. PMID- 11484732 TI - JAMA patient page. Schizophrenia. PMID- 11484733 TI - Economics of coronary artery bypass grafting. 1985. PMID- 11484736 TI - [One flew over the cardiologist's nest...]. PMID- 11484734 TI - Dyspnea and pulmonary infiltrates in a 53-year-old woman with common variable immunodeficiency. PMID- 11484737 TI - The Second Chance Club. PMID- 11484738 TI - Heritable bone disease and stroke due to vertebral artery dissection. PMID- 11484739 TI - Are all surgeons, patients and procedures created equal? PMID- 11484740 TI - Leak-point pressure: clinical application of values at two different volumes. AB - A prospective analysis of 306 consecutive patients with genuine stress incontinence was performed to evaluate the clinical usefulness of additional leak point pressure (LPP) determination at 200 ml. LPP values at both volumes were compared to maximal urethral closure pressure (MUCP) in an attempt to determine a critical cut-off value for the detection of a low MUCP (< or =20 cmH2O). A positive LPP at 150 ml was found in 157 patients. The mean LPP for patients with a low MUCP was 58.5 cmH2O compared to 71.6 for those with a normal MUCP, which was statistically significant (p = 0.01). The correlation coefficient between LPP and MUCP was 0.317. A negative LPP was found in 30% (24/79) of the total having a low MUCP. The addition of values for LPP at 200 ml resulted in an increase in the number who leaked to 191, a 50% increase in the detection rate of low MUCP and a statistically significant relationship between LPP < or =60 cmH2O and low MUCP. Various critical cut-off values for LPP demonstrated good specificity but poor sensitivity for the detection of a low MUCP. It was concluded that there was a statistically significant relationship between LPP and MUCP. Performing LPP at 200 ml provides additional clinically useful diagnostic information. PMID- 11484741 TI - Can uroflowmetry patterns in women be reliably interpreted? AB - Our aim was to investigate the proposition that uroflowmetry patterns can be reliably interpreted and correspond with specific urodynamic diagnoses. Uroflowmetry traces from 129 women with diagnoses of either genuine stress incontinence or detrusor instability were interpreted by four physicians with a minimum of 6 months experience in urogynecology. To test intraobserver variability, the traces were classified a second time 8 weeks later. Inter- and intraobserver variability was calculated by kappa analysis. There was marked intra- and interobserver variability in classification of traces, but no evidence of a correlation between urodynamic diagnosis and uroflowmetry pattern. Neither peak flow, total voided volume nor rate of acceleration of flow correlated with diagnosis. Although flow rates are important in predicting possible problems following surgery for stress incontinence, there is no evidence that flow patterns can be used as a screening test for specific urodynamic diagnoses. PMID- 11484742 TI - Power Doppler of the urethra in continent or incontinent, pre- and postmenopausal women. AB - Urethral pressure should exceed bladder pressure, both at rest and on stress, for urinary continence to occur. A decrease in urethral pressure is a major factor explaining the pathogenesis of urinary incontinence. A number of elements, such as smooth and striated periurethral muscles, and connective, vascular and elastic tissues, contribute to urethral pressure. The periurethral vessels are influenced by hormonal changes during the menstrual cycle, during pregnancy and postmenopause. We studied the periurethral vessels in 97 women, 57 of whom were incontinent and 40 continent, using power color Doppler velocimetry. The number of periurethral vessels, systolic peak, minimum diastolic values, pulsatility and resistance indexes, as well as systolic-diastolic ratio, were assessed. Statistically significant differences were found between incontinent women in the premenopausal period and those in the postmenopausal period, regarding the number of periurethral vessels, systolic peak, minimum diastolic values, pulsatility and resistance indexes. PMID- 11484743 TI - Bladder augmentation techniques in women. AB - Augmentation cystoplasty is performed in women for a variety of indications, including neurogenic bladder dysfunction, inflammatory diseases, small fibrosed bladders, idiopathic urge incontinence and enuresis. The preoperative evaluation, surgical techniques and postoperative management are reviewed. Complications of stone formation, urinary tract infections, mucus production, metabolic disturbances, hematuria and dysuria syndrome, tumors and perforations are addressed. In addition, alternative techniques such as ureterocystoplasty, autoaugmentation, seromuscular cystoplasty and the future of augmentation of the bladder utilizing techniques of tissue engineering are discussed. The management of pregnancy in women who have previously undergone augmentation cystoplasty is also reviewed. PMID- 11484744 TI - Perioperative complications of Burch colposuspension. AB - In a review of the literature on the perioperative complications of Burch colposuspension we found only sparse data associated with this problem. We describe our own experience (> 1800 procedures) and the literature, discussing bleeding/hematoma, injury to the bladder, kinking/injury to the ureters, voiding dysfunction, infection, and rare complications such as deep venous thrombosis, pulmonary embolism, ileus, sepsis, external iliac vein injury and mortality rate. Knowledge of the possible risks and complications of Burch colposuspension may minimize the intraoperative complications and increase postoperative surgical success and patient satisfaction. PMID- 11484745 TI - Use of the artificial urinary sphincter in women. AB - Persistent urinary incontinence after failed surgical repair can be successfully treated with the artificial urinary sphincter. The English literature was reviewed from 1985 to 1996. Eleven articles and abstracts addressing placement of the artificial urinary sphincter in women were identified. Discussion includes two operative techniques. Success rates were in the range of 91%-99%. Erosion rates were 7%-29%. The artificial urinary sphincter is an effective treatment for women failing other procedures. Appropriate work-up and diagnosis for type III stress urinary incontinence is crucial. PMID- 11484746 TI - Urinary diversion for incontinence in women. AB - Urinary incontinence is a common condition affecting up to 50% of the female population, but only a third seek medical help. Although the majority of these are satisfactorily managed with conservative or conventional surgical treatment, 10%-15% continue to remain incontinent despite intervention. Urinary diversion is a management option for this group. The different techniques of urinary diversion, their results, complications and long-term sequelae are discussed, with a review of the literature. PMID- 11484747 TI - A critical view on the value of urodynamics in non-neurogenic incontinence in women. AB - The requirements for reliable urodynamics are standardized techniques, including uniform pressure sensors, filling rates, position and posture during the investigation, and uniform diuresis. Physiological variations in flow and urethral pressure profile (UPP) (menstrual cycle, intensity of coughing, circadian variations) must be considered. Parameters of the UPP (maximum (closure) urethral pressure, pressure-transmission ratio and leak-point pressure) are useful if interpreted with caution. Uninhibited detrusor contractions are more frequently recorded in ambulatory urodynamics, and range from 'subthreshold' to very strong. No quantification formulae correlate with subjective symptoms or degree of urge (incontinence). Mixed incontinence can make the results of surgery worse, but do not so necessarily. Postoperative dysuria cannot be predicted from urodynamics, as surgical factors are more important. Electromyography is not useful in non-neurogenic female incontinence. For routine nonneurogenic incontinence extensive urodynamic testing can be reduced to one pressure measurement; more complicated cases must be tested by a physician with large practical experience and a theoretical background. PMID- 11484748 TI - Vaginal leiomyoma--an imitator of prolapse. AB - Vaginal leiomyoma is a rare tumor with a variable clinical presentation and broad differential diagnosis that can lead to preoperative misdiagnosis. We present a case of vaginal leiomyoma with a symptom complex of prolapse, urinary urgency and urge incontinence. A 50-year-old woman presented with a 4-year history of deteriorating sensation of prolapse, significant complex urinary complaints and prolonged vaginal bleeding. Clinical examination revealed a mobile 6 x 8 cm mass arising from the anterior vaginal wall. She underwent hysteroscopy, curettage, urethrocystoscopy (normal findings) and mass enucleation through a vertical incision. Histology showed a benign leiomyoma. Ultrasonography, MRI, positive pressure urethrography and urethrocystoscopy should be considered in the evaluation of an anterior wall vaginal mass. Surgical enucleation via a vaginal approach is the treatment of choice. If this surgical procedure results in skeletonization of the urethral and bladder support, a colporrhaphy/pubourethral ligament plication is required. PMID- 11484749 TI - Pregnancy following tension-free vaginal taping. AB - Genuine stress incontinence is usually managed conservatively in women desirous of further childbearing. Tension-free vaginal taping is a relatively new method of surgical treatment of genuine stress incontinence. We report a case of pregnancy following the procedure. The patient was delivered by elective cesarean section. She remained continent throughout pregnancy and on a 6-month postnatal follow-up. PMID- 11484750 TI - Percutaneous peripheral revascularisation with excimer laser: equipment, technique and results. AB - Laser angioplasty has been evaluated for coronary applications since the early 1980s. Early complications of dissection, perforation and thermal injury led to a loss of enthusiasm for this technique. Recent advances in catheter development, including optimally spaced laser fibres, athermic 308 nm wave-length catheters, and saline infusion techniques have produced larger laser channels, minimised thermal injury and significantly reduced vessel dissection. This improvement in equipment and technique has led to the growing use of lasers in the field of percutaneous treatment of peripheral vascular interventions. Convincing data supporting laser use in thrombus may lead to widespread use of laser in diffuse, thrombotic, long occlusions in the SFA (superficial femoral artery) and for infrapopliteal disease treatment in patients with non-healing ulcers. Little literature exists on the specific results of 308 nm Excimer laser catheter use for peripheral angioplasty. Significant research is still needed to prove the role of debulking in peripheral applications, but upcoming clinical trial data from the PELA (peripheral angioplasty vs. laser study in long SFA occlusions) and LACI (laser angioplasty in chronic ishaemia) may help to solve these questions in the near future. This article attempts to outline the technical issues of laser catheter use in percutaneous peripheral interventions, including access, sheath selection and wire techniques to cross even the most challenging obstructions in the peripheral circulation. PMID- 11484751 TI - Application of excimer laser coronary angioplasty (ELCA) in bifurcation lesions. AB - Excimer laser coronary angioplasty (ELCA) offers a unique approach to the treatment of bifurcation lesions that continues to present a challenge in percutaneous coronary intervention. Debulking plaque prior to stenting or balloon angioplasty has demonstrated significant improvement in the treatment of bifurcation lesions. Clot dissipation properties of excimer laser combined with its ability to debulk, makes this device unique when applied to thrombus-laden bifurcation lesions. ELCA is the only debulking technique that allows retention of two guide wires with resultant protection of the bifurcation vessels during the debulking procedure. We herein describe three patients with unstable angina who underwent a debulking procedure with ELCA involving application of single and double wire techniques. These three cases illustrate two different techniques used for debulking bifurcation lesions. Both single and double wire techniques are described. ELCA can be used safely and effectively in high-risk patients with bifurcation lesions, even in the presence of thrombus. PMID- 11484752 TI - Excimer laser for pacemaker and defibrillator lead extraction: techniques and clinical results. AB - Pacing and defibrillation leads may need to be removed for several reasons including infection, interference with others leads, lack of vascular access or redundancy. However, the removal of chronically implanted leads is a major technical challenge because of the extensive adhesions that develop along the course of the leads over time. The techniques to remove chronic leads have been greatly facilitated by the development of an excimer laser sheath. We undertook an analysis of our experience with laser extraction in the first 50 leads attempted. An excimer laser sheath system, developed by the Spectranetics Corporation, was used to extract 50 chronically implanted leads in 34 patients. The mean patient age was 64+/-12 years, all were male and the average duration that the leads had been implanted was 5.0+/-3.9 years. Two-thirds of the leads were pacemaker and one-third were defibrillator leads. There was a 100% clinical success rate and 48 of the 50 leads were completely removed. There were no major complications. There was one minor complication of subclavian vein thrombosis and two haemodynamically non-significant episodes of air embolism. The main limitation observed was failure of the excimer laser sheath to advance in 18% of cases, probably due to the presence of calcified adherences on leads. Two strategies were found useful to deal with this problem: under the clavicle stainless-steel sheaths were used to break up calcified adherences and within the venous system the laser sheath was upsized in order to advance over the calcification on the lead. It was concluded that excimer laser has greatly facilitated the removal of chronically implanted pacemaker and defibrillator leads. There is a high success rate and low complication rate in our experience. The main limitation of laser is the presence of calcified adherences. PMID- 11484753 TI - Current multicentre studies with the excimer laser: design and aims. AB - Excimer laser ablation to remove atherosclerotic plaque has been used for over a decade as a methodology to treat cardiovascular disease. Improvements in the technique and technology of excimer laser angioplasty, coupled with the recognition of new clinical opportunities for this treatment modality, have resulted in a resurgence of interest in the laser. Three clinical trials are now being conducted to explore potential applications, including the LARS trial of excimer laser versus balloon angioplasty to treat in-stent restenosis, the PELA trial of excimer laser angioplasty in occluded superficial femoral artery disease, and the LACI trial of excimer laser angioplasty in limb-threatening ischaemia. This article describes the rationale and objectives of these new approaches to some of the more challenging problems in cardiovascular disease. PMID- 11484754 TI - Excimer laser thrombolysis: an emerging option for acute ischaemic coronary syndromes. PMID- 11484755 TI - Next generation catheters for excimer laser coronary angioplasty. AB - In response to the need for maximising debulking in complex lesions, three new excimer laser coronary angioplasty catheter designs have been introduced. The eccentric laser catheter features a fibreoptic bundle disposed opposite the guide wire lumen at the catheter tip and a torque mechanism that allows the user to rotate the fibre bundle toward the lesion mass. Residual lumens 50% larger than the catheter tip diameter have been obtained when multiple passes were made, with each pass performed using a different tip rotation. A recent case series utilising this catheter in restenosed stents resulted in larger lumens and lower 6-month restenosis rates. The optimal spaced (OS) laser catheter features a fibre bundle placed concentrically around the guide-wire lumen. The 61 microm diameter core fibres are spaced at a nominal centre-to-centre distance of 90 microm, resulting in a 40% increase in ablative area as compared to previous concentric catheter designs. In vitro testing and clinical evaluation demonstrated OS catheters routinely achieve an ablated area > or =90% of the catheter tip size. The 0.9 mm catheter features a high-density fibre pack composed of 65 fibres. Peripheral dead space has been minimised to maximise penetration of calcified plaque. When combined with laser parameters of up to 80 mJ/mm2, and 80 Hz pulse repetition rate, the catheter demonstrated improved hard tissue and calcified tissue penetration in vitro. Clinical evaluation in Canada revealed a 94% lesion recanalisation rate in high-grade stenoses with angiographic evidence of calcification, chronic total occlusions, and lesions which have failed balloon angioplasty. PMID- 11484756 TI - A model for evaluation of arterial thrombosis following interventional procedures. AB - A model to assess thrombus formation following vascular injury was evaluated using various interventional systems. The model consisted of a 'stretchable' shunt box that served as an arteriovenous shunt between the carotid artery and jugular vein in dogs. Arterial homografts obtained from both carotid and femoral arteries were mounted between two plastic connectors attached to either side of the shunt box. The opposing walls of the shunt box were then stretched apart to achieve the original length of the arteries. The arterial side of the box was connected to the ipsilateral carotid artery and the venous side was connected to the contralateral jugular vein. Haemostasis valves were placed at the exit ports on the venous side of the shunt box. These were used as an access to the various interventional catheters into the lumens of the homografts. Interventions were performed prior to initiating blood flow. After the interventions, 111indium labelled platelets were injected on the arterial side of the shunt box and arterial blood flow initiated across the shunt. After one hour of circulation through the shunt box, the blood flow was interrupted, and the homografts were perfusion-fixed with glutaraldehyde and segments removed for radioactive counts and processed for histology. This shunt box was then used to compare platelet adhesion and thrombus formation after balloon angioplasty (BA) to direct laser (LA) and laser-thermal angioplasty (LTA). A total of 28 arteries were used from seven dogs. In each experiment, one homograft was used as control, and three other homografts were treated with either BA, LA or LTA. Following the interventions, 111indium-labelled platelets were injected and circulated for one hour using the dog's native circulation. Labelled platelet counts for BA (19102+/ 4869/cm2; mean +/- SE) were significantly greater than LA (7038+/-980/cm2), thermal LTA (5189+/-1961/cm2), and control (1575+/-541/cm2), respectively (p<0.05, ANOVA). Histology examination showed few platelets at LA, LTA and control sites whereas extensive platelet adhesion was noted at BA treated sites. The model provided a means to conduct simultaneous comparison of several interventions under similar conditions. In this case thermal treatment of the arterial homografts had the least amount of platelet adhesion. PMID- 11484757 TI - Excimer laser revascularisation: current indications, applications and techniques. AB - The ultraviolet pulsed excimer laser (308 nm wavelength) is currently the only laser approved by the FDA for percutaneous intervention in patients with ischemic coronary artery disease. The clinical presentation of the treated patients varies from stable and unstable angina to acute myocardial infarction. Potential advantages of excimer laser revascularisation in acute coronary syndromes and in ischaemic obstructive peripheral vascular disease include concomitant plaque debulking and thrombus removal; absence of systemic lytic state; shortened thrombus clearing time and facilitation of adjunct balloon angioplasty and stenting. Improved understanding of laser-tissue interactions and positive clinical outcomes through the use of safe lasing techniques have led to expansion of indications/applications for laser angioplasty. These include stent restenosis, complex lesions and thrombotic stenoses, bifurcation lesions, balloon failure, total occlusions, focal saphenous vein graft lesions and peripheral arterial obstructions. The excimer laser can be effectively utilised in patients with depressed left ventricular ejection fraction and does not require implantation of a temporary pacemaker as no-reflow phenomenon and severe arrhythmias are rarely encountered. Careful case selection, proper utilisation of equipment and incorporation of efficient lasing techniques play a crucial role in effective and safe cardiovascular laser applications. PMID- 11484758 TI - Excimer laser for revascularisation of saphenous vein grafts. AB - Coronary artery bypass grafting has been a major advance in cardiovascular medicine over the past 30 years. Saphenous venous bypass grafts, however, are prone to develop atherosclerotic disease within several years of the procedure. Unfortunately, percutaneous interventional techniques in saphenous venous bypass grafts are associated with significant risks of distal embolisation and resultant non-Q wave myocardial infarction. Preliminary results suggest excimer laser angioplasty may significantly reduce this complication. This article summarises the results of percutaneous interventions in spahenous venous bypass grafts, emphasising the potential role of excimer laser angioplasty in this group of patients. PMID- 11484759 TI - Excimer laser coronary angioplasty (ELCA) for diffuse in-stent restenosis: beneficial long-term results after sufficient debulking with a lesion-specific approach using various laser catheters. AB - Although brachytherapy became frequently used technique for treating in-stent restenosis (ISR) in 2000 (stenosis recurrence: 14-25%), removing the tissue (i.e., debulking) is still under evaluation. Regarding XeCl Excimer laser no randomised trials have been completed so far. The entirely attainable and published data originate from five one-centre and one multicentre registries which showed a high procedural success-(98%) and a very low complication rate (<2%), but heterogeneous results based on (I) manifold laser- and lasing technology due to evolving catheter and lasing technology during the studies and (II) inhomogeneous patient-populations, which had been documented only occasionally, although we now know that ISR-recurrence rates are significantly higher in patients with (1) high grade stenosis (>70%), (2) long lesions (>15mm in length), (3) recurrent ISR. (4) ISR in saphenous vein grafts, (5) early-ISR (<3 months after stent implantation), (6) multi-vessel disease and (7) low ejection fraction. Because all athero-ablative procedures are typically finalised by adjunctive balloon PCI, diameter stenosis before adjunctive PTCA was between 23% and 64%, reflecting the different lasing (and therefor debulking) intensity between the studies. The mean lesion-length, vessel diameter, stent-type and coronary differed significantly between the studies (i.e. mean lesion length between 14 and 32 mm) and although of major influence, the speed of lasing (=ablation) was not recorded in the various studies. Concerning eccentric ELCA the invention of the area-ablation technique provide a significant greater ablation effect and the new OS-technology provide 60% more ablating area. In summary, the feasibility, effectiveness, procedural success and presumably long term outcome of the Excimer laser for treating ISR depend on multiple outcome influencing aspects. The lack of recording variables make analysis of various laser-for-ISR trials difficult or even impossible. ELCA seems to have advantageous procedural and long term results in lesions, where aggressive debulking effect using eccentric ELCA catheters can be achieved. The OS-catheter can provide a maximum debulking in distal lesions and tortuous vessels, where the debulking abilities of the eccentric catheter are limited. If laser debulking is inadequate (DS 30%), the incidence of recurrent restenosis appear to be higher and brachytherapy is probably more suitable. Sufficient laser debulking can only be achieved through use of precise technique respecting the laser-specific abilities of plaque dissolution, appropriate energy levels and quantities, and possibly recovery time. PMID- 11484760 TI - Excimer laser (308 nm) recanalisation of in-stent restenosis: thermal considerations. AB - Excimer laser recanalisation of in-stent restenosis may be a viable modality for improving coronary patency. However, the presence of arterial stents modifies the thermal properties of the irradiated area and may alter temperature patterns generated during ablation. The goal of this study was to evaluate, in vitro, temperature changes during excimer laser ablation of stented vessels and compare them with those obtained from unstented (control) vessels. Six different stent types (AVE Microstent-II, AVE-GFX, ACS Multi-link, JJ Palmaz-Schatz, JJ Crown, and NIR) were deployed in freshly excised porcine coronary vessels. Three control unstented samples were also measured. Blood or saline was infused through the vessels, while the tissue environment was kept at approximately 37 degrees C. A 308 nm excimer laser (Spectranetics, CVX300) with an eccentric 2.0 mm laser catheter (Spectranetics, EII) delivered two trains of 200 pulses each, 10 s apart, at 60 mJ/mm2, and 40 Hz, simulating maximum clinical exposure. The catheter was positioned midway in the stent, first coaxially parallel to the vessel wall, and then at an angle against the stent and vessel wall. Temperature measurements (n= 168 for blood, n=96 for saline) were performed with a approximately 210 microm diameter, fast-response thermocouple with 0.1 degrees C resolution. The probe was positioned to within approximately 250 microm from the inner surface of the vessels. Tissue temperature was measured at the catheter tip and at the distal and proximal edges of the stents. Maximum recorded temperatures for coaxial and angular alignment, did not exceed 42.2 degrees C (approximately 6 degrees C above baseline) and 54.2 degrees C (approximately 18.1 degrees C above baseline) respectively, for all stents types tested, controls, and all probe locations. Both stented and unstented vessels exhibited comparable temperature gradients. The observed maximum temperatures, obtained under extreme lasing conditions, indicated that 308 nm ablation, in the presence of stents under blood or saline infusion, produces clinically acceptable temperatures. PMID- 11484761 TI - Protein kinase C and phosphatidylinositol 3-kinase independently contribute to P glycoprotein-mediated drug secretion in the mouse proximal tubule. AB - We have recently reported that the apical membrane of the proximal tubule S2 segment from wild-type mice (WT mice) has the capacity for P-glycoprotein- (P-gp ) mediated drug efflux, whereas mice in which both mdr1a and mdr1b genes are disrupted (KO mice) do not. To examine the intracellular regulatory mechanisms of drug-transporting P-gp activity, we isolated and perfused proximal tubule S2 segments from WT and KO mice, and measured luminal efflux of the intracellular fluorescence of rhodamine 123, a fluorescent substrate of P-gp. The decay half time of the intracellular fluorescence (t1/2) was regarded as an index of the drug-transporting P-gp activity. In the WT mice, the t1/2 was 36+/-5 s (n=35) during the basal period, and was significantly increased to 440+/-45 s by the luminal addition of verapamil (an inhibitor of P-gp). The addition of phorbol 12 myristate 13-acetate (PMA) [a protein kinase C (PKC) activator] to the bath increased t1/2, but 4alpha-phorbol (the inactive form of PMA) did not. The PMA induced increase in t1/2 was further increased by the luminal addition of verapamil, and was partially inhibited by co-treatment with staurosporine or H-7 (inhibitors of PKC). Pretreatment with wortmannin or LY294002 [inhibitors of phosphatidylinositol 3-kinase (PI 3-kinase)] added to the bath also increased t1/2. The wortmannin- and LY294002-induced increase in t1/2 was also further increased by the luminal addition of verapamil. The effects of PMA on t1/2 were additive after cotreatment with wortmannin or LY294002. In the KO mice, t1/2 was 440+/-25 s (n=18) during the basal period, and was no longer affected by the addition of verapamil, staurosporine, H-7, wortmannin, or LY294002. Based on the use of pharmacological agents, we conclude that in the proximal tubule from WT mice, P-gp-mediated drug secretion occurs independently via PKC- and PI 3-kinase dependent processes, whereas it is not present in KO mice, independently of PKC- and PI 3-kinase. PMID- 11484762 TI - Inhibition of cardiac delayed rectifier K+ currents by an antisense oligodeoxynucleotide against IsK (minK) and over-expression of IsK mutant D77N in neonatal mouse hearts. AB - The IsK (minK or KCNE1) protein is known to co-assemble with the KvLQT1 (KCNQ1) protein to form a channel underlying the slowly activating delayed rectifier K+ current (IKs). Controversy remains as to whether the IsK protein assembles with ERG (the ether-a-go-go-related gene) products to form or modulate the channel underlying the rapidly activating delayed rectifier K+ current (IKr). We investigated the effects of antisense oligodeoxynucleotides (AS-ODN) against IsK and its mutant D77N [which underlies a form of long QT syndrome (LQT5) in humans] on the delayed rectifier K+ current (IK) of neonatal mouse ventricular myocytes in primary culture. Patch-clamp experiments on these cells showed that IK consists of IKs and IKr. IK was not recorded from ventricular cells transfected with AS-ODN, while it was recorded from cells transfected with the corresponding sense oligodeoxynucleotides (S-ODN). IK was not recorded from cells transfected with the D77N mutant, and the action potential duration was much longer than in cells transfected with wild-type IsK. Furthermore, HERG could not induce currents in COS-1 cells co-expressed with the D77N mutant and HERG (the human form of ERG). These results indicate that the IsK protein associates with both KvLQT1 and ERG products to modulate IKr and IKs in cardiac myocytes. PMID- 11484763 TI - Platelet activating factor increases ciliary activity in the hamster oviduct through epithelial production of prostaglandin E2. AB - We investigated the signal transduction mechanisms associated with an increase in ciliary beat frequency (CBF) produced by platelet activating factor (PAF) in oviductal ciliated cell cultures. In the range of concentrations similar to that produced by preimplantation embryos, PAF increased the CBF in a dose-dependent manner. The addition of PAF and prostaglandin E2 (PGE2) to the cultures produced a synergic increase of ciliary beating, suggesting that PAF and PGE2 signal transduction pathways may be associated. To demonstrate this hypothesis, cyclooxygenase-2 (COX-2) was selectively blocked by a specific inhibitor, NS-398, and the PAF-induced CBF increase was abolished. Moreover, a phospholipase A2 (PLA2) inhibitor, AACOCF3, blocks the PAF-induced CBF increase. PGE2 production by oviductal epithelial cells is stimulated by PAF, and WEB-2086, a PAF-receptor blocker, specifically blocks the PAF-induced PGE2 production. Using the fluorescent indicator fura-2, we measured the effect of PAF on intracellular Ca2+ concentration ([Ca2+]i) in individual ciliated cells. PAF induced a transient increase of [Ca2+]i that was blocked by WEB-2086 or by removal of extracellular Ca2+. We propose a mechanism for PAF-mediated signal transduction in the ciliated cells of the oviductal epithelium. Minimal doses of PAF trigger Ca2+ mobilization in tandem with increased PLA2 activity and a COX-2-mediated increase in PGE2. Local PGE2 production by the oviductal mucosa suggests the presence of an autocrine loop controlling ciliary activity. PMID- 11484764 TI - Alpha2-adrenoceptors inhibit antidiuretic hormone-stimulated Na+ absorption across tight epithelia (Rana esculenta). AB - In the present study we examined the effect of alpha-adrenergic regulation of active transepithelial Na+ absorption across the isolated frog skin epithelium. alpha-Adrenergic stimulation was achieved by addition of the adrenergic agonist noradrenaline in the presence of the beta-adrenergic blocker propranolol. alpha Adrenergic stimulation inhibited basal as well as antidiuretic hormone (ADH) stimulated Na+ transport. The ADH-induced increase in Na+ transport was accompanied by a membrane depolarisation due to an increase in the apical Na+ permeability. The subsequent application of noradrenaline inhibited the Na+ transport and repolarised the membrane potential, suggesting that alpha adrenergic stimulation had reduced the apical Na+ permeability. The inhibition was abolished by the alpha2-adrenergic antagonist yohimbine whereas it was insensitive to the alpha1-adrenergic antagonist prazosin. alpha-Adrenergic stimulation had no effect on the cytosolic free [Ca2+] ([Ca2+]i). Incubation of the epithelium in the presence of ADH increased the cellular adenosine 3',5' cyclic monophosphate (cAMP) content, an increase which was abolished by alpha adrenergic activation. The effect of alpha-adrenergic stimulation on cAMP production was abolished by the alpha2-adrenergic antagonist yohimbine. We conclude that the noradrenaline-induced inhibition of the ADH-stimulated Na+ absorption and cAMP content is mediated by activation of alpha2-adrenoceptors. The data further indicate that the principal cells of the epithelium do not express alpha1-adrenoceptors. The noradrenaline-induced inhibition of the ADH stimulated Na+ transport was concentration dependent, with 0.24+/-0.03 microM eliciting a half-maximal response. This alpha2-adrenergic-mediated down regulation of Na+ absorption is achieved at a concentration of noradrenaline which begins to activate the NaCl secretion via the skin glands. The alpha2 adrenoceptors therefore appear to have considerable physiological importance. PMID- 11484765 TI - Compensatory role of CaMKII on ICa and SR function during acidosis in rat ventricular myocytes. AB - It has been suggested that the activity of Ca2+/calmodulin-dependent protein kinase II (CaMKII) increases during acidosis in cardiac muscle. Thus we have investigated the role of CaMKII during acidosis by monitoring intracellular Ca2+ (using fura-2) and ICa (using the perforated patch clamp technique) during acidosis, in the absence and presence of the CaMKII inhibitor KN-93, in rat isolated ventricular myocytes. In the absence of KN-93, acidosis (pH 6.5) increased the amplitude of the fura-2 transient and prolonged its decay, but in the presence of KN-93 acidosis did not alter the amplitude and prolonged the decay more. In the absence of KN-93, acidosis increased the amplitude of the caffeine-induced fura-2 transient but did not alter its amplitude in the presence of KN-93. ICa did not change significantly during acidosis in the absence of KN 93 but decreased during acidosis in the presence of KN-93. These results suggest that activation of CaMKII during acidosis helps to compensate for the direct inhibitory effects of acidosis on sarcoplasmic reticular Ca2+ uptake and ICa. PMID- 11484766 TI - Water drinking causes a biphasic change in blood composition in humans. AB - To investigate precisely the fluid shifts associated with water drinking in humans, we measured continuously blood density and plasma electrolyte concentrations using the mechanical oscillator technique and ion-selective electrodes, respectively, in healthy young volunteers before (10 min) and after (48 min) water drinking for a period of 2 min. Beat-by-beat blood pressure was also monitored throughout the experiment. Drinking 1 l tap water caused a transient increase in blood density immediately after the drinking episode (from 1051.1+/-0.5 g/l before drinking to 1051.8+/-0.5 g/l 4 min after the start of drinking, P<0.05), followed by a gradual reduction (1050.1+/-0.5 g/l at 31 min). This drinking-induced change paralleled those of haematocrit, plasma density and plasma volume. Plasma [Na+] and [Cl-] and osmolality decreased after drinking without transient increases and reached minima at about 30 min. A transient increase in mean arterial blood pressure was observed prior to the increase in blood density. These findings suggest that water drinking causes a biphasic change in plasma volume: initial haemoconcentration, probably due to sympathetic acceleration, followed by haemodilution due to the post-absorptive effect, and further suggest that the fluid shift associated with the initial haemoconcentration is isosmotic. PMID- 11484767 TI - Adult dystrophic (mdx) endplates exhibit reduced quantal size and enhanced quantal variation. AB - Examination of miniature endplate potential (MEPP) distributions indicated that the average quantal size is significantly reduced by approximately 44% in young adult mdx mice (5-7 weeks). The average quantum declined further to approximately 37% of non-dystrophic levels in mature (6- to 24-month) mdx mice. Young adult non dystrophic and mdx endplates and mature non-dystrophic endplates exhibited a linear relationship between the mean and variance of uniquantal MEPP amplitude distributions. Mature mdx endplates, however, exhibited a distinctly nonlinear relationship characterized by large increases in variance at larger mean MEPP amplitudes. These results indicate a reduced average density of functional acetylcholine (ACh) receptors (AChRs) at mdx endplates that, in mature preparations, is associated with greater temporal or spatial variability in the density of functional AChRs apposed to individual release sites. PMID- 11484768 TI - Intracellular calcium mobilization by muscarinic receptors is regulated by micromolar concentrations of external Ca2+. AB - Carbachol-induced contractions of rat stomach fundus strips, obtained in a nutrient solution containing 1.8 mM Ca2+, were resistant to Ca2+ withdrawal, even after 1 h of bathing the tissues in a nominal 0 Ca2+ solution. This was not observed when K+ was used to evoke contractions, which were rapidly inhibited after Ca2+ removal (t1/2=2 min). The effect of carbachol in 0 Ca2+ solution was reduced by using drugs that reduce intracellular pools of Ca2+, such as caffeine (1-3 mM), ryanodine (30 microM) or thapsigargin (1 microM), corroborating the involvement of intracellular Ca2+ stores. On the other hand, when the 0 Ca2+ solution contained EGTA, a complete decline of carbachol effects was observed within about 8 min, indicating the involvement of extracellular Ca2+. Atomic absorption spectrometry showed that our 0 Ca2+ solution still contained 45 microM Ca2+, which was drastically reduced to 5.9 nM in the presence of EGTA. Taken together, our results indicate that the effects of carbachol are due to the mobilization of caffeine-, ryanodine- and thapsigargin-sensitive intracellular Ca2+ stores, and that these stores are not inactivated or depleted if micromolar concentrations (45 microM), but not nanomolar concentrations (5.9 nM) of Ca2+ are maintained in the extracellular milieu. PMID- 11484769 TI - Ontogenesis of leptin expression in different adipose tissue depots in the rat. AB - Serum leptin levels and leptin mRNA expression by adipose tissue increase with age and are mainly associated with an increase in adiposity. Regional changes in both leptin production and fat distribution contribute to circulating leptin levels and may play a role in the regulation of body weight. a capacity that changes during development. Here, we have studied leptin mRNA expression in four different white adipose tissue depots (epididymal, retroperitoneal, mesenteric, inguinal; namely, EWAT, RWAT, MWAT, IWAT) and in interscapular brown adipose tissue (IBAT). We have also studied their relationship with lipid content and adiposity changes, together with serum leptin levels in male rats at different ages (18, 55, 93, 159, 212, 294 and 355 days). Serum leptin levels increased during development, reaching stable levels at the age of 7 months, and, as expected, were highly correlated with both the adiposity index (r=0.908, P<0.01) and body weight (r=0.906, P<0.01). Leptin mRNA expression also increased with age, following characteristic ontogenic patterns in every adipose tissue depot. The patterns were similar in EWAT and RWAT: leptin expression increased very rapidly during the first 55 days for EWAT and 3 months for RWAT, with a peak in the latter at 7 months, and high expression levels were retained for the rest of the study period. In IWAT and IBAT, leptin expression increased steadily during the 12-month period studied and was significantly lower than levels in EWAT and RWAT. Leptin expression in MWAT increased progressively with age to reach levels close to those of EWAT and RWAT in 10-month-old animals. The pattern of leptin expression in both EWAT and RWAT paralleled their lipid content, and leptin mRNA expression per unit of tissue lipid content was maintained high and constant from a very young age (about 2 and 3 months, respectively). However, the expression of mRNA for leptin (expressed per unit of tissue lipid concentration) in MWAT, IWAT and IBAT increased steadily during the whole period studied, without attaining the maximal levels observed in EWAT and RWAT. MWAT, IWAT and IBAT maintained their capacity to increase leptin mRNA expression in response to an additional accumulation of lipids. Our data demonstrate that there are regional-specific differences and different rates of increase of leptin gene expression within distinct depots of WAT and BAT. These changes cannot be uniquely explained by changes in adiposity or lipid content, implying that there are regional-specific regulatory mechanisms that may depend on the attenuation with age of the beta adrenergic inhibitory signalling pathway upon leptin expression or on other factors. PMID- 11484770 TI - Effects of phosphatidylinositol kinase inhibitors on the activation of the store operated calcium current ICRAC in RBL-1 cells. AB - In electrically non-excitable cells, Ca2+ entry is mediated predominantly through the store-operated Ca2+ influx pathway, which is activated by emptying the intracellular Ca2+ stores following an increase in the levels of the second messenger inositol 1,4,5-trisphophate (InsP3). InsP3 is generated from the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2). Recently, roles for other phosphoinositides (PIs) in store-operated Ca2+ influx have been suggested because inhibitors of PI kinases reduce Ca2+ influx when the latter is triggered independent of PIP2 hydrolysis. Using the whole-cell patch-clamp technique to record the store-operated Ca2+ current ICRAC in RBL-1 cells, we examined whether PIs are involved in linking store depletion to activation of CRAC channels. Of several structurally distinct PI kinase inhibitors, only one (LY294002) was able to reduce partially the extent of activation of ICRAC although this could not be reversed by exogenous phosphatidylinositol 3,4,5 trisphosphate (PIP3). Our findings suggest that, if a PI kinase is involved in activation of ICRAC in RBL-1 cells, it has a unique pharmacological profile. Alternative explanations for the results are discussed. PMID- 11484771 TI - Sexual dimorphism in the adrenergic control of rat brown adipose tissue response to overfeeding. AB - Gender-related differences in the brown adipose tissue (BAT) response to overfeeding rats on a cafeteria diet were studied by assessing the balance between the expression of beta-adrenoceptors (beta1-, beta2-, beta3-AR) and alpha2A-AR and their relation to the expression of uncoupling proteins (UCP1, UCP2, UCP3). Cafeteria diet feeding for 15 days, which involved a similar degree of hyperphagia in both sexes, led to a greater body weight excess in females than in males and a lower activation of thermogenesis. Gender-related differences were found for different adrenoceptor expression and protein levels, which might explain, in part, sex differences in the thermogenic parameters. The lower expression of alpha2A-AR in females than in males could be responsible for the higher expression of UCP1 and thermogenic capacity under non-hyperphagic conditions. However, in a situation of high adrenergic stimulation--as occurs with overfeeding--as there is a preferential recruitment of the beta3-AR by noradrenaline compared with other adrenergic receptors, the higher levels of beta3-AR in males rats than in females could be responsible for the greater thermogenic capacity and the lesser weight gain in males. Thus, the alpha2/beta3 balance in BAT could be a key in the thermogenic control. PMID- 11484772 TI - Evidence that glutamate acting on presynaptic type-II metabotropic glutamate receptors alone does not fully account for the phenomenon of depolarisation induced suppression of inhibition in cerebellar Purkinje cells. AB - Depolarisation-induced suppression of inhibition (DSI) is a form of short-term synaptic plasticity at gamma-aminobutyric-acid-(GABA)ergic synapses between principal neurons and interneurons in both the cerebellum and the hippocampus. The induction of DSI involves an intracellular calcium-dependent release of a retrograde messenger from the postsynaptic principal neuron (Purkinje cell/pyramidal cell in cerebellum/hippocampus) onto presynaptic interneurons, where it is thought to bind to guanine nucleotide-binding protein (G protein) coupled receptors and subsequently reducing GABA release from these interneurons onto the postsynaptic principal neuron. Pharmacological studies have indicated that glutamate might be a retrograde messenger in both cerebellum and hippocampus, where, in the former at least, it seems to activate type-II metabotropic glutamate receptors (mGluRs). Using LY-341495, a recently described, highly specific and potent antagonist of type-II mGluRs, to block these receptors reduced DSI slightly, but significantly, in spite of the fact that this antagonist completely suppressed the effects of stimulating type-II mGluRs with a specific agonist. Activation of type II mGluRs alone thus cannot account fully for DSI in cerebellum and hence other mechanisms are involved in its induction. Such mechanisms probably involve an additional retrograde signal. PMID- 11484773 TI - Diphenylamine-2-carboxylic acid (DPC), Usually an inhibitor of Cl- and non selective cation channels, inhibits Cl-/HCO3- exchange and opens Cl- and cation conductances in rabbit gallbladder epithelium. AB - In the apical plasma membrane of rabbit gallbladder epithelium various drugs (hydrochlorothiazide, phlorizin, phenylglyoxal) inhibit Cl-/HCO3- exchange and probably enhance the almost negligible intrinsic anion conductance of the exchanger. By radiochemical measurements of apical Cl- influx, the anion exchange is shown here to be directly and immediately inhibited by diphenylamine-2 carboxylic acid (DPC) too. Using conventional microelectrode techniques in intact tissue, DPC, with same dose/response curve, is shown to activate an apical anion conductance (GCl) that has similar properties and amplitude to the GCl activated by the other exchange inhibitors so far tested; the actions are not additive. Patch-clamp methods (cell-attached and excised inside-out patch configurations) reveal that GCl is due to anion channels that are non-rectifying, cytoplasm independent, sensitive to stilbene and dipyridamole and have conductance of a few picosiemens. All this strengthens the correlation between inhibition of anion exchange and the activation of GCl and channels with features similar to those of the almost negligible intrinsic anion conductance of the exchanger. Among the drugs tested, the effects of DPC and hydrochlorothiazide are even more similar, such that even their dose/response curves overlap. Moreover, both drugs also directly activate some verapamil-sensitive Ca2+ channels and consequently apamin sensitive, Ca2+-activated K+ channels. Thus DPC, usually an inhibitor of Cl- and non-selective cation channels, is shown here to be capable of activating Cl- and cation conductances. PMID- 11484774 TI - Effect of endurance training on oxidative and antioxidative function in human permeabilized muscle fibres. AB - The adaptation of muscle oxidative function to 6 weeks of endurance cycle training was investigated in eight untrained subjects. Peak oxygen consumption (VO2peak) increased by 24% (2.69+/-0.21 versus 3.34+/-0.30 l O2 min(-1), mean +/ SEM, P<0.01) and lactate threshold intensity increased by 53% (121+/-13 versus 185+/-15 W, P<0.01) following the training period. Muscle biopsy samples were taken from vastus lateralis before and after training, and respiration in permeabilized muscle fibres was measured. Following training, non-ADP-stimulated respiration (VO) of skinned fibres increased by 35% (0.17+/-0.01 versus 0.23+/ 0.01 mmol O2.min(-1).kg(-1) wet weight, P<0.05) and maximal ADP-stimulated respiration (VmaX) increased by 38% (1.17+/-0.07 versus 1.62+/-0.14 mmol O2.min( 1).kg(-1) wet weight, P<0.05). ADP sensitivity [i.e. the ratio between mitochondrial respiration (after correction for VO) at 0.1 mM ADP and Vmax] was reduced after training (0.40+/-0.05 versus 0.26+/-0.02; P<0.05). Mitochondrial resistance to oxidative stress was investigated by exposing skinned fibres to exogenous reactive oxygen species (ROS). ADP-stimulated respiration was reduced after ROS exposure and the relative decrease was similar before and after training. It is concluded that after endurance training: (1) the relative increase in maximal muscle fibre respiration exceeds that of whole-body oxygen uptake; (2) the sensitivity of mitochondrial respiration to ADP decreases; and (3) the impairment of oxidative function in skinned muscle fibres by ROS remains unchanged. PMID- 11484775 TI - Superoxide dismutase gene expression is activated by a single bout of exercise in rat skeletal muscle. AB - The goal of this experiment was to examine contraction-mediated activation of superoxide dismutase (SOD) gene expression in rat superficial vastus lateralis (SVL, type IIb) and deep vastus lateralis (DVL, type IIa) muscles. Female Sprague Dawley rats were randomly divided into exercise (E) and control (C) groups that were sacrificed at 0, 1, 2, 4, 10, 24, and 48 h (n=6) following an acute bout of treadmill exercise (25 m/min, 5% grade) to exhaustion (running time approximately equals 1 h). Nuclear factor-kappaB (NF-kappaB) in DVL and SVL showed maximal binding at 2 and 10 h respectively, and remained elevated. Activator protein-1 (AP-1) showed maximal binding at 1 h post-exercise, and returned to resting levels at 10 h in both muscles. Mn SOD mRNA abundance in the DVL was increased at 0 (P<0.01), 1, and 2 h (P<0.05) post-exercise, whereas Mn SOD protein was unchanged. In SVL, Mn SOD mRNA abundance was not altered by exercise, whereas Mn SOD protein content was increased at 10 (P<0.05) and 24 h (P<0.075) post exercise. CuZn SOD mRNA was unchanged with exercise in DVL and SVL, but CuZn SOD protein was elevated 48 h after exercise in both DVL and SVL (P<0.01). Activities of Mn SOD, CuZn SOD and total SOD showed no change with exercise in either muscle examined. These findings indicate that an acute bout of exercise can increase binding of NF-kappaB and AP-1 in both SVL and DVL, which may stimulate Mn SOD mRNA transcription in the more oxidative type DVL muscle. The increased CuZn SOD protein contents seen post-exercise, without increases in mRNA abundance in both DVL and SVL, suggest a translational mechanism in this SOD isoform. PMID- 11484776 TI - Staircase-like potentiation of calcium release in mouse myotubes during repetitive short-term application of threshold caffeine. AB - The release of Ca2+ in response to caffeine at threshold concentration (5 mM) was studied in mouse skeletal myotubes. Repeated 5-s applications of caffeine, each followed by a 30-s washout, caused Ca2+ releases of consecutively growing amplitude (staircase phenomenon). Each response declined rapidly and had a slow tail. Repeated applications of threshold caffeine lowered the threshold concentration. The interval between threshold applications could be increased to 30 min without loss of potentiation. When threshold caffeine was applied continuously for up to 10 min, the increase in Ca2+ concentration as seen with staircase potentiation did not occur. Depolarization by elevated [K+] or by voltage-clamp steps potentiated caffeine-induced Ca2+ release rapidly as compared to the slow exponential growth of staircase-like potentiation. Gd3+ prevented the depolarization-induced potentiation, but not the staircase phenomenon. Staircase like potentiation of Ca2+ release was evident even when the voltage sensors were clamped in their resting state; in contrast, potentiated Ca2+ release and its rapid termination apparently require conversion of the voltage sensors to an activated state. Staircase potentiation was blocked when Ca2+ was omitted from the bath, thus pinpointing the source of Ca2+. We suggest that staircase-like potentiation is conditioned by a caffeine-dependent Ca2+ influx across the plasma membrane. PMID- 11484777 TI - Oxygen delivery and oxygen return in humans exercising in acute normobaric hypoxia. AB - At a given steady O2 consumption (VO2) in normoxia, cardiac output (Q) is inversely proportional to arterial O2 concentration (CaO2), so that O2 delivery (QaO2=QCaO2) is kept constant and adapted to VO2. The matching between QaO2 and VO2 keeps O2 return (QvO2=QaO2-VO2) constant and independent of VO2 and haemoglobin concentration ([Hb]). This may not be so in hypoxia: in order for QvO2 to be independent of the inspired O2 fractions (FIO2), the slopes of the Q versus VO2 lines should be greater the lower the CaO2, which may not be the case. Thus, we tested the hypothesis of constant QvO2 by determining QaO2 and QvO2 in acute hypoxia. Thirteen subjects performed steady-state submaximal exercise on the cycle ergometer at 30, 60, 90 and 120 W breathing FIO2 of 0.21, 0.16, 0.13, 0.11 and 0.09. VO2 was measured by a metabolic cart, Q by CO2 rebreathing, [Hb] by a photometric technique and arterial O2, saturation (SaO2) by infrared oximetry. CaO2 was calculated from [Hb], SaO2 and the O2 binding coefficient of haemoglobin. The VO2 versus power relation was independent of FIO2. The relations between Q and VO2 were displaced upward and had higher slopes in hypoxia than in normoxia. However, the Q changes did not compensate for those in CaO2. The slopes of the QaO2 versus VO2, lines tended to decrease in hypoxia. QVO2 was lower the lower the FIO2. A significant relationship was found between QvO2 and SaO2 (QvO2= 1.442 SaO2+0.107, r=0.871, n=24, P<10(-7)), which confutes the hypothesis of constant QvO2 in hypoxia. PMID- 11484778 TI - Cholinergic input to the supraoptic nucleus increases Fos expression and body temperature in rats. AB - To examine the role played by cholinergic input and processes in the supraoptic nucleus (SON) in the control of body temperature and water intake in rats, we used microdialysis to stimulate and analyze SON without disturbing the behavior of unanesthetized rats. After microdialysis, we also investigated immunoreactivity for c-Fos protein in the brain as an index of neuronal activation. Stimulation with neostigmine, an acetylcholine esterase inhibitor, through the microdialysis probe increased the extracellular concentration of acetylcholine in the SON. This cholinergic stimulation dose-dependently increased body temperature but did not significantly change the water intake. The stimulation markedly increased c-Fos-like immunoreactivity (Fos-IR) in the SON and certain hypothalamic areas, including the paraventricular nucleus (PVN) and median preoptic nucleus (MnPO). Fos-IR was also evident in certain regions of the pons and brainstem, including the locus ceruleus (LC), area postrema (AP), and nucleus of the solitary tract (NTS). Addition of atropine, a muscarinic receptor antagonist, to the dialysis medium containing neostigmine attenuated the increase of Fos-IR and suppressed the neostigmine-induced responses in body temperature. These results suggest that cholinergic input and activation of the muscarinic cholinoceptive neurons in the SON contribute to the regulation of body temperature. Activation of noradrenergic pathways in the brainstem including LC and NTS may be involved in the thermoregulation mechanism. PMID- 11484779 TI - pH regulation and buffering power in gastric smooth muscle. AB - Intracellular pH can have profound effects on tissue function, but little is known about how pH is regulated, buffered or affects the function of gastric smooth muscle. As the pH of gastric myocytes may alter with pathophysiological disturbance of the gastric lining, or reduction in blood flow to the stomach, these parameters were investigated. Intracellular pH was measured in strips of corpus from rats and guinea-pigs and pH perturbed by the addition of Na butyrate. pH regulation was investigated using pharmacological inhibitors and ionic substitutions. Resting pH was found to be around 7.0, and buffering power relatively high, compared to other muscles in both species. In the guinea-pig amiloride, EIPA and HOE694 prevented pH regulation from an acid load, but amiloride- and EIPA-insensitive pH-regulating mechanisms were found in the rat. The pH-regulatory mechanism present in the rat was also insensitive to DIDS, SITS and removal of external Cl-, but inhibited by Na+ substitution and HOE694. Acidification reduced gastric tone in both species. We conclude that pH alteration will significantly affect gastric contractility, despite a high capacity of the tissue to buffer and regulate pH change. The sensitivity to NHE inhibitors differs between rat and guinea-pig, suggesting that Na+/H+ exchanger isoform expression differs between gastric tissue. PMID- 11484780 TI - Phosphocreatine and ATP content in human single muscle fibres before and after maximum dynamic exercise. AB - The recovery of high-energy phosphate levels in single human skeletal muscle fibres following short-term maximal (all-out) exercise was investigated. Three male volunteers exercised maximally for 25 s on an isokinetic cycling ergometer. Muscle biopsy samples from the vastus lateralis were collected at rest, immediately post-exercise and at 1.5 min of recovery. The subjects also performed a second exercise bout 1.5 min after the first, on a separate occasion. Single muscle fibres were dissected, characterized and assigned to one of four groups according to their myosin heavy chain (MyHC) isoform content; namely, type I, IIA, IIAx and IIXa (the latter two groups containing either less or more than 50% IIX MyHC). Fibres were analysed for adenosine 5'-triphosphate (ATP), inosine-5' monophosphate (IMP), phosphocreatine (PCr) and creatine (Cr) levels. Type I fibres had a lower Cr content than type II fibres (P<0.01). Within type II fibres resting [PCr] increased with increasing MyHC IIX isoform content (r=0.59, P<0.01). Post-exercise [PCr] was very low in all fibre groups (P<0.01 versus rest) while great reductions in ATP were also observed (P<0.01 versus rest), especially in the type II fibre groups. [PCr] at 1.5 min of recovery was still lower compared to rest for all fibre groups (P<0.01) especially in the IIAx and IIXa fibres. PMID- 11484781 TI - Identification of processes governing long-term accumulation of 137Cs by forest trees following the Chernobyl accident. AB - The regularities of 137Cs distribution in trees (Pinus sylvestris and Betula pendula) growing in different types of forest ecosystems were investigated. High levels of heterogeneity of 137Cs activity concentrations in different parts of the trees, resulting from their varied metabolism have been shown. The data obtained demonstrate a non-uniform character of 137Cs distribution along the trunks, which can be explained by radionuclide fixation by the xylem vessel walls and by geometry changes along the tree trunk. It has been found that the radial distribution of 137Cs in the tree trunk is dependent on the availability of 137Cs in soil, which governs the transfer of this radionuclide via xylem sap and on the properties of the xylem. The accumulation of 137Cs by trees was influenced by the vertical distribution and availability of 137Cs in the soil as well as by the root biomass distribution in different soil horizons. A bioavailability factor, which takes into account the vertical distribution of radiocesium in soil, bioavailability of this radionuclide and distribution of root biomass in different soil horizons is proposed for comparative analyses of 137Cs transfer from soil to trees in different types of forest ecosystems. PMID- 11484782 TI - Diurnal and seasonal variation of the equilibrium state between short-lived radon decay products and radon gas in ground-level air. AB - To study seasonal and diurnal variations and the effect of meteorological parameters, the equilibrium factor F (i.e. the ratio of equilibrium equivalent radon daughter concentration and radon gas concentration) was determined as a result of measurements on a test field at Munich-Neuherberg, Germany, continuously from October 1995 through March 1997. On average, F was found to be 0.62+/-0.09 (95% confidence level). The time series of F showed no distinct seasonal variations. Nevertheless, typical diurnal variations as well as seasonal variations of the diurnal behaviour were observed. Generally, F was found to be increased in the early afternoon, i.e. under conditions of enhanced vertical mixing in the atmosphere. The daily differences between high and low values of F depended on the season. On average, low F values were characteristic for days with precipitation and high wind speed, i.e. under turbulent atmospheric conditions. Therefore, taking daily mean values into account, F was found to be positively correlated with the aerosol concentration, although a relationship between the diurnal behaviour of the aerosol concentration and that of F was not detectable. PMID- 11484783 TI - Cytogenetic biomonitoring carried out in a village (Dolon) adjacent to the Semipalatinsk nuclear weapon test site. AB - The Semipalatinsk region (Kazakhstan Republic) has been affected by extensive radioactive contamination due to more than 450 nuclear tests of which almost 100 were exploded in the atmosphere. The present results refer to cytogenetic assessments in a study cohort of the population of Dolon, a settlement located on the NE boundary of the nuclear weapon test site, which was exposed to elevated doses of ionising radiation primarily due to the first Soviet nuclear test in 1949. Conventional cytogenetic analyses were carried out on 21 blood samples from individuals (more than 50 years old) living in Dolon since the very beginning of nuclear testing. A matched control group included 20 individuals living in non contaminated areas. Higher frequencies of chromosome aberrations were found in the Dolon cohort compared to the control group, even though they remain within the range of the background levels reported for large normal human population studies on elderly individuals. PMID- 11484784 TI - Age dependence of T-lymphocyte apoptosis induced by high-energy proton exposure. AB - Peripheral blood samples from three donors of different ages were exposed to 300 kVp x-rays or 138 MeV protons (0.2, and 9 Gy dose). After 48 h incubation, CD4 and CD8 T-lymphocytes were labelled with specific monoclonal antibodies and cellular DNA was stained by propidium iodine. Radiation-induced apoptosis was followed by flow cytometry and the data were processed by LYSIS II software. The data analysis revealed an age-dependent sensitivity to radiation-induced apoptosis by 300 kVp x-rays and 138 MeV protons, for both CD4 and CD8 T lymphocytes. Radiation-induced apoptosis was about 4 times greater in CD4 lymphocytes from the youngest donor than the oldest donor and was about 2 times greater in CD8 T-lymphocytes, both after x-ray and proton exposures. RBE values for CD4 T-lymphocytes ranged from 0.9 to 1.4 and for CD8-positive cells from 0.7 to 0.9. It is concluded that radiation-induced apoptosis of CD4 and CD8 T lymphocytes, which is already exploited to predict patient response in conventional radiotherapy, may also be used to predict response in proton treatment planning. PMID- 11484785 TI - Caffeine-inhibitable control of the radiation-induced G2 arrest in L5178Y-S cells deficient in non-homologous end-joining. AB - The two L5178Y (LY) sublines bear a heterozygous Tp53 mutation that affects its transactivation function. LY-S (radiation-sensitive) cells are deficient in double strand break (DSB) repair by non-homologous end-joining (NHEJ) and do not express p21WAF1 (Cdkna1) either constitutively or after x-irradiation, in contrast to their radiation-resistant counterpart LY-R cells, which express p21WAF1 constitutively. Radiation-induced G2 arrest in LY-S cells is very long (11 h/Gy) but 2 mM caffeine treatment shortens it, decreases the fraction of G2 cells and increases the fraction of apoptotic cells. The treatment also increases the DNA damage that is estimated with the comet assay 18 h after irradiation with 5 Gy (ca. 23% of the initial value for x-rays and ca. 47% for x-rays plus caffeine). This indicates that either the repair has not been completed or the apoptotic DNA fragmentation has been initiated (or both). The same treatment applied to x-irradiated (5 Gy) LY-R cells (G2 arrest, 4 h/Gy) has no radiosensitising effect, induces no apoptosis and does not alter the amount of DNA damage left unrepaired (ca. 28%). The results are compatible with the assumption that inhibition of the Atm-dependent homologous recombination repair by caffeine, brings differential effects in LY sublines because of the defect of the alternative DNA repair system (NHEJ) in LY-S cells. PMID- 11484786 TI - Photobiological activity of exogenous and endogenous porphyrin derivatives in Escherichia coli and Enterococcus hirae cells. AB - Photodynamic treatment, the combined application of a photosensitiser and visible light, represents a new and promising approach for the inactivation of microorganisms. The photosensitising potentials of exogenous zinc-phthalocyanine tetrasulphonate (ZnPsTS), tetraphenylporphyrins (TPPs) and endogenous porphyrin derivatives were tested and compared on Gram-negative and Gram-positive bacteria, Escherichia coli B. and Enterococcus hirae, respectively. The synthesis of endogenous porphyrins was induced by 5-aminolevulinic acid (delta-ALA). The porphyrin- or delta-ALA-treated cells were irradiated with white light. The photosensitising efficiency of endogenous derivatives on both types of bacteria is ZnPcTS < TP(4-OGluOH)3P < TP(4-OGluOH)4P. However, neither exogenous derivatives exhibit appreciable photosensitising activity for disinfection application. ALA-induced photodynamic treatment showed good potential for the inactivation of Escherichia coli cells, but not towards Enterococcus hirae cells. The failure of photosensitisation of the Enterococcus hirae strain selected indicates that apart from the Gram-positive character, other structural elements of the membrane can influence the result of photodynamic treatments. PMID- 11484787 TI - The adult male voxel model "Golem" segmented from whole-body CT patient data. AB - This paper describes the construction of an adult male voxel model named "Golem" intended to be used for Monte Carlo simulations to calculate dosimetric quantities for radiation protection considerations. The model was segmented from whole-body medical image data of a living person who was 38 years old and had external dimensions close to those of the ICRP Reference Man. The segmentation process using dedicated image processing hard- and software is described and the resulting model is characterised with respect to weight and height of the total body, organ and tissue masses and red bone marrow distribution. A comparison with the respective data for ICRP Reference Man and three further voxel models is presented. Golem was found to agree reasonably well with Reference Man, so that he can be used for the assessment of "representative" body doses. PMID- 11484788 TI - Bowman-Birk proteinase inhibitor-mediated radioprotection against UV irradiation is TP53-dependent and associated with stimulation of nucleotide excision repair. AB - The Bowman-Birk proteinase inhibitor (BBI) has previously been described as a radioprotective agent against ionising radiation. It was demonstrated that BBI is able to significantly increase the clonogenic cell survival of normal fibroblasts when applied before exposure to ultraviolet B (UVB) radiation. In transformed TP53-mutated cell lines, however, the BBI-mediated radioprotection was absent. At the molecular level, the radioprotective effect of BBI can be correlated with BBI mediated stabilisation of TP53 protein prior to irradiation. Following UVB irradiation, the BBI-treated cells present an accelerated removal of cyclobutane pyrimidine dimers. Thus, the cell and molecular biological data presented suggest that BBI is able to protect cells with functional TP53 from UVB-induced DNA damage. This protective effect is most likely achieved via the activation of the TP53 signalling cascade resulting in the activation of nucleotide excision repair. PMID- 11484789 TI - Is there a real risk of radiation-induced breast cancer for postmenopausal women? AB - The risk of radiation-induced breast cancer decreases with increasing age at exposure. Thus, for calculating the individual risk for a patient undergoing mammography, age-related risk coefficients need to be used. In this report, the results of epidemiological studies on risks of radiation-induced breast cancer are reviewed indicating that the available data do not show the risk to be enhanced for women exposed at the age of 55 years or older. This lack of evidence is reflected by the fact that the risk coefficients recommended by national and international advisory bodies differ by a factor of 10 or more for age at exposure of 50-60 years or older. A hypothesis is proposed indicating that the risk of radiation-induced breast cancer might decrease considerably at the time of menopause. The hypothesis is based on the following line of arguments: (1) evidence has accumulated from molecular genetic studies indicating that the development of colorectal cancer requires a cascade of subsequent mutations consisting of at least seven genetic events. (2) For colorectal cancer, the annual rates of incidence and mortality increase with age to the power of 5-6. Thus, the number of mutation steps (minus 1) is approximately reflected by the power of age dependence. (3) For western populations, the incidence and mortality of breast cancer up to the age of about 50 years increase with age to the power of about 6, indicating that a similar number of genetic events might be involved in development of breast cancer as has been identified for colorectal cancer. (4) For women aged 50 years or older, breast cancer occurs at an annual rate that is about proportional to age or age squared. This may mean that after menopause, the processes in the multistep mutation cascade leading to breast cancer are slowed down by a factor of about 4 or more. (5) The constant relative risk model of radiation carcinogenesis implies for solid cancers that radiation acts by inducing additional mutations in the earlier steps of the multistep cascade. It is suggested that the break-point in the age-specific annual rate of breast cancer incidence at menopause is associated with a corresponding drop in radiation sensitivity with respect to induction of breast cancer. PMID- 11484790 TI - Considerations on the behavior of long-lived radionuclides in the soil. AB - The migration of radionuclides from waste repositories to the biosphere potentially leads to a contamination of soil. Due to the importance of food production, the mobilisation and accumulation behaviour of long-lived radionuclides in the soil plays a key role in performance assessment studies. In this paper, the main features and processes that control radionuclide behaviour in soil, such as pH, redox potential and sorption to organic and inorganic soil components, are discussed for the radionuclides 36Cl, 79Se, 129I, 99Tc, 237Np and 238U, that are usually most relevant in long-term safety assessments of nuclear waste. The interaction of radionuclide behaviour in soil with environmental factors, such as temperature and humidity as well as farming practices are discussed. The possible impact of future soil development on long-term behaviour in soil are taken into consideration. Due to the physiological constraints of plant growth, appropriate soil conditions for growth will probably not be substantially different from current requirements, bearing in mind that sustainable agriculture strives for optimal plant growth. Against this background, present-day parameters may in general be considered appropriate for roughly estimating the behaviour of radionuclides in the soil-plant system. PMID- 11484791 TI - Digital imaging modalities for dental practice. AB - The introduction of the computed tomograph in the 1970s revolutionized medical diagnosis by initiating the transition from analogue to digital imaging. During this period, more specialized equipment for image processing was developed, such as cathode-ray tubes for image display, special sensors for image acquisition, and storage devices for image archiving. Digital imaging systems designed exclusively for use in dentistry were developed in the latter half of the 1980s. Some are now being clinically applied under conditions of close scrutiny to determine diagnostic accuracy, image quality, and radiation exposure to patients. This article reviews the enabling technologies of digital systems used in dentistry, and focuses upon intraoral digital imaging systems, concepts for digital image acquisition, and variations in radiation dose and their effects on diagnostic accuracy of caries detection. PMID- 11484793 TI - Application of sintered titanium alloys to metal denture bases: a study of titanium powder sheets for complete denture base. AB - The purpose of this study was the fabrication of titanium powder sheets to enable the application of sintered titanium alloys as metal denture bases. The effects of titanium particle shape and size, binder content, and plasticizer content on the surface smoothness, tensile strength and elongation of titanium powder sheets was investigated. To select a suitable ratio of powdered metal contents for application as a metal denture base, the effects of aluminum content in Ti sheets and various other powder metal contents in Ti-Al sheets on the density, sintering shrinkage, and bending strength were evaluated. Based on the results of the above experiments, we developed a mixed powder sheet composed of 83Ti-7Al-10Cr with TA45 titanium powder (atomized, -45 microm), and 8 mass% binder content. This titanium alloy sheet had good formability and ductility. Its sintered titanium alloy had a density of 3.2 g/cm3, sintering shrinkage of 3.8%, and bending strength of 403 MPa. The titanium alloy sheet is clinically acceptable for fabricating denture bases. PMID- 11484792 TI - The identification of the sympathetic neurons innervating the hamster submandibular gland and their electrophysiological membrane properties. AB - The neuron innervating the hamster submandibular (SM) gland was identified in the superior cervical ganglion (SCG) in vitro by recording the antidromic response using the intracellular recording technique. After the cellular response was recorded, methylene blue was injected iontophoretically into the neuron from the recording electrode, and the location of the cell soma was determined. The salivatory neurons of the SM gland were in the small- to medium-sized group of the entire cell population of the SCG. The cell size was 36.3 x 24.4 microm (mean, n=45). The postganglionic fibers were entirely unmyelinated (mean: 0.34 m/sec at 28-30 degrees C, n=141). Eighty-seven percent of the cells were distributed in the central one-third of area between the external carotid nerve origin and the caudal pole in the SCG. The resting membrane potential, membrane input resistance, membrane time constant and membrane input capacitance of the salivatory neuron were as follows: -49.2+/-7.6 mV (n=102), 52.9+/-23.6 Mohms (n=71), 8.0+/-3.4 msec (n=71) and 147+/-50 pF (n=71). Fast- and slow-excitatory postsynaptic potentials (EPSPs) were evoked, but not slow-inhibitory postsynaptic potentials (IPSPs). The fast EPSP was 13.1+/-5.7 mV in amplitude and 46.2+/-17.1 msec in duration (n=35). The slow EPSP (20 Hz, 5 sec) was 6.9+/-11 .9 mV in amplitude and 101+/-43 sec in duration (n=16). The directly-evoked spike was 63.0+/-11.9 mV in amplitude and 5.9+/-1.3 msec in duration (n=54). The spike after-hyperpolarization (AHP) was 12.5+/-3.5 mV in amplitude and 353+/-161 msec in duration. Na+ and Ca+ channels were involved in the spike generation. The voltage-dependent K+ channels (delayed rectifier), A channels and rapidly Ca2+ activated K+ channels (BK channels) regulated the spike-falling phase. The delayed rectifiers, A channels, and BK and SK (slowly Ca2+-activated) channels were involved in generation of spike-AHP. Muscarine suppressed the Ca2+ component of spike via muscarinic receptors. PMID- 11484794 TI - A case of amelogenesis imperfecta of deciduous and all permanent teeth. AB - We experienced a case with severe enamel defects of both the deciduous teeth and all the permanent teeth. In order to clarify the etiology of enamel defects in this patient, we performed a DNA analysis in addition to conventional examinations. Although we suspected a variety of systemic factors causing enamel defects, there was no evidence suggesting disturbances of amelogenesis. In the present case, we suspected a mutation in the amelogenin gene and performed nucleotide sequencing of the exons of the amelogenin gene, but we could not find any evidence of mutation. We suggest that a mutation of some other gene related to enamel formation or the adventitious factors contributed to the amelogenesis imperfecta in this case. PMID- 11484795 TI - A case of an ameloblastic fibro-odontoma arising from a calcifying odontogenic cyst. AB - This case report describes an ameloblastic fibro-odontoma arising from a calcifying odontogenic cyst (COC) in the mandible of a twenty-three-year old male. The patient was referred to the Department of Oral Surgery, Tokyo Dental College, on March 30th, 2000, complaining of a painful swelling, which had appeared three weeks earlier on his left mandibular molar region. In a pathological view, the lesion was a round cyst the size of a chicken-egg, dark red in color, and surrounded by a thick membrane. The cyst had an epithelium of varying thickness which included many ghost cells and an enamel-like structure on the inside, and a thick wall of connective tissue with an ameloblastic fibro odontoma on the outside. Enamel organ-like epithelial islands were structured radially in the form of strands with immature dentin. Cytokeratin 19 was strongly immunoreactive in the epithelium of the lesion; osteopontin and osteocalcin reacted in the mesenchymal cells and weakly in the epithelial element of this tumor. PMID- 11484796 TI - Stability and change in structural social relations as predictor of mortality among elderly women and men. AB - In a follow-up study of 70-95 years old women and men (n = 911) we studied the association between change and stability in three structural aspects of social relations (contact frequency, contact diversity, cohabitation status) from 1986 1990 and mortality after the next four years in 1994. Women aged 70-74 years who developed low contact frequency or developed small contact diversity showed significantly higher mortality, adjusted ORfreq: 3.78 (1.08-13.20), adjusted ORdiv: 3.79 (1.24-11.58). Women aged 70-74 years with continuously low contact frequency showed an increased mortality compared to women constantly experiencing high contact frequency, adjusted OR: 2.75 (1.04-7.26). A tendency in the same direction for sustained small contact diversity was found, adjusted OR: 1.98 (0.70-5.61). Among women aged 75+ years no impact of frequency and diversity was demonstrated, whereas continuously living alone was a significant predictor of mortality, when compared to women continuously living with somebody, adjusted OR: 2.57 (1.29-5.09). In men, we found a significantly increased mortality among those who developed high contact frequency and developed large contact diversity ORfreq: 3.91 (1.02-14.94) and ORdiv: 6.04 (1.30-28.03). In summary, we found rather larger age differences in the strength of the association between change in structural social relations and mortality. Furthermore, the associations seemed stronger among women than men, which may however mainly be explained by the small number of men in our cohort. PMID- 11484797 TI - Low prevalence of Chlamydia pneumoniae and Mycoplasma pneumoniae among patients with symptoms of respiratory tract infections in Dutch general practices. AB - Acute respiratory disease is one of the most common reasons to consult a general practitioner. A substantial part of these diseases cannot be explained by an infection with a virus or a common pathogenic bacterium. To study this diagnostic deficit, the prevalence of Chlamydia pneumoniae and Mycoplasma pneumoniae infections was determined in two groups of patients consulting a general practitioner. DNA of C. pneumoniae and M. pneumoniae was detected by a polymerase chain reaction (PCR) in nose/throat swabs from six (1.1%), and from seven (1.3%) patients, respectively, of 557 patients consulting a general practitioner for complaints suggestive for a virus infection during the 1994/1995 respiratory infections season. Two patients remained C. pneumoniae PCR-positive for at least 4 weeks. All others were negative within 3 weeks. Double infections of C. pneumoniae and influenza virus (3/6), and of M. pneumoniae and respiratory syncytial virus (1/7) or rhinovirus (1/7) were diagnosed. During the 1992/1993 season, attempts to isolate C. pneumoniae in cell culture or to detect C. pneumoniae DNA by PCR using throat swabs were all negative for 80 patients with a sore throat, although serological data suggested a C. pneumoniae infection in 13 (16%) patients. A specimen from another patient of this group was M. pneumoniae PCR-positive and the corresponding serum specimens showed a persistent high antibody titre. In summary, the prevalence of acute C. pneumoniae and M. pneumoniae infections was less than 2% in patients consulting a general practitioner. PMID- 11484798 TI - High frequency of the HIV-1 protective CCR5 delta32 deletion in native Estonians. AB - In previous studies, the highest frequencies (16%) of the CCR5 delta32 deletion have been found in populations of Finno-Ugric origin. We here report a high CCR5 delta32 frequency (15%) in another Finno-Ugric populations, the Estonians. The highest frequency (18%) was found on the geographically isolated Estonian island of Dago. We examined 504 healthy unrelated individuals of Estonian nationality, whose grandparents were born in Estonia. The polymerase chain reaction assay was performed and the amplified products were digested with EcoRI. PMID- 11484799 TI - Space-time clustering of childhood lymphatic leukaemias and non-Hodgkin's lymphomas in Sweden. AB - BACKGROUND: The discussion concerning clusters of childhood leukaemia has mainly been focused on their relation to the time and place of diagnosis. Recently some studies have indicated clustering not only at diagnosis, but also around time and place of birth. Space-time clustering at time of birth could be of special interest if the aetiological agent is of infectious origin and the induction of leukaemia either occurs pre- or perinatally or an infection at that time favours a poor subsequent immune response to the agent. METHODS: To identify possible space-time clustering we have used the close-pair method of Knox. One-thousand twenty recorded cases (0-14 years) of childhood acute lymphatic leukaemia and 293 cases (0-14 years) of malignant non-Hodgkin's lymphoma from Sweden between 1973 1996 were analysed. The records include date of birth and of diagnosis as well as addresses at birth and at diagnosis. RESULTS: A significant excess of case-pairs (25 observed, 14.9 expected, p = 0.01) was observed close in date and place of birth in the 4-14 year age group with acute lymphatic leukaemia (ALL). However there was no statistically significant clustering found around time of diagnosis. When the cases of leukaemia and the non-Hodgkin's lymphomas were combined no statistically significant clustering was obtained neither at birth nor at diagnosis. CONCLUSIONS: This study strengthens the evidence of space-time clustering around the birth date in children whom later developed ALL. This observation is in support of the hypothesis that pre- or perinatal infections can induce a process leading to ALL. PMID- 11484800 TI - Is there an association between passive smoking and breast cancer? AB - Recent epidemiological studies have suggested passive smoking as possible risk factors in breast cancer etiology. However, the evidence is not conclusive. We conducted a meta-analysis of the risk of breast cancer associated with passive smoking. Eleven studies were identified through Medline, Cancer Abstracts and abstract presented at related scientific meetings. Combined estimates of relative risks (RR) were calculated using fixed and random effect models. Significant heterogeneity was detected among the studies (chi2 = 34.6, p < 0.01). The combined estimate of RR for ever exposure to passive smoking was 1.41 (95% confidence interval (CI): 1.14-1.75). The combined estimate for published studies was 1.43 (95% CI: 1.10-1.85). Only two studies reported significant dose-response relation between level of passive smoking and premenopausal breast cancer. Our analysis suggests a possible weak association between passive smoking and breast cancer. More studies are needed to establish whether the observed association is causal. PMID- 11484801 TI - Comparison of balanced and random allocation in clinical trials: a simulation study. AB - BACKGROUND: Before analysing the results of a randomised controlled clinical trial in which 200 children were balanced over five prognostic factors, we wanted to know the efficiency of balanced allocation compared to simple randomisation as well as the efficiency of adjusted as compared to unadjusted statistical analysis in small and large sample sizes. METHODS: A simulation study with 1000 replications of each assignment was performed for both relatively large trials (n = 100) and for small trials (n = 20). Four options for the design and analysis were studied: (1) simple randomisation with simple univariate analysis, (2) simple randomisation with multivariate modelling, (3) balanced allocation with simple univariate analysis and (4) balanced allocation with multivariate modelling. In addition, we also considered the effect of an unmeasured covariable, which was either uncorrelated or correlated with another covariate. RESULTS/CONCLUSION: The simulations show that a combination of balanced allocation and multivariate analysis as compared to simple randomisation and multivariate analysis leads to more valid and precise treatment effects as well as to smaller confidence intervals, especially in small trials (n = 20). Multivariate analysis with all known prognostic factors produces on average smaller Type I errors and Type II errors in balanced allocation compared to simple randomisation. If an unmeasured covariate is strongly correlated with another covariate the treatment effect is estimated more precisely as compared to an unmeasured covariate that is not correlate or less strongly correlated. PMID- 11484802 TI - Moderate to severe iodine deficiency in three endemic goitre areas from the Black Sea region and the capital of Turkey. AB - Endemic goitre is still an important and underestimated health concern in Turkey. The overall prevalence had been calculated as 30.3% by palpation in a national survey conducted in 1995. However, direct evidence that iodine deficiency (ID) is the major cause of the endemic were lacking until now. We measured sonographic thyroid volumes (STV), urinary iodine concentrations (UIC) in 1226 school age children (SAC) (9-11 year old) from Ankara the capital of Turkey located in the central Anatolia, and three highly endemic goitre areas of the Black Sea region. A considerable number of school age children (SAC) were found to have STV exceeding the recommended upper normal limits for their age and gender obtained from iodine-replete European children (i.e. 26.7, 40.3, 44.8 and 51.7% of children from Ankara, Kastamonu, Bayburt and Trabzon respectively). UIC indicated moderate to severe ID in these areas with median concentrations of 25.5, 30.5, 16.0 and 14 microg/L respectively. This study showed severe to moderate ID as the primary etiological factor for the goitre endemic observed in Ankara and the Black Sea region of Turkey. PMID- 11484803 TI - A change-point model for reporting delays under change of AIDS case definition. AB - Accurate monitoring of disease incidence is of major public health concern. The time delay between diagnosis and the date of reporting creates bias in estimating disease incidence. Changes in case definition are expected to have an impact on the time lag of case reporting. We propose a change-point model for reporting delays in AIDS that takes into account recent changes in the AIDS definition in US and European countries. The model was applied to California AIDS surveillance data and the distribution of reporting delays before and after the recent change of definition in 1993 were analyzed in terms of contributing factors. The overall significance of the model with change-point as compared to the model without change-point indicates that the effect of the 1993 change in definition on the distribution of reporting delays was highly significant (p < 0.0001). Overall, reporting delay of cases initially diagnosed with AIDS-defining diseases before 1993 was shorter compared to after 1993; reporting delay of cases initially diagnosed meeting the 1993 immunologic case definition was shorter than of those initially diagnosed with AIDS-defining diseases. Region of residence, mode of exposure, race/ethnicity and time of diagnosis emerged as the main covariates in the models. The method introduced here applies to current and possible future changes of the AIDS case definition as well as changes in diagnostic criteria or case definition in diseases other than AIDS. We demonstrate that such changes may be accompanied by sizeable changes in the distribution of reporting delays, and thus adjustment for reporting delays must be recalibrated after a change in definition. PMID- 11484804 TI - Urbanisation and vitamin A deficiency in children: comparison between a traditional district and a new settlement in Mali. AB - OBJECTIVES: The purpose of this study was to assess the vitamin A status of pre school urban children and to compare the situation between a traditional district and a new settlement after two decades of intense urbanisation. SUBJECTS AND METHODS: A cross-sectional stratified survey was carried out in Bamako, Mali, on a representative sample of children (aged 4-6 years). 532 children were involved from a traditional district and 453 from a new settlement (NS). The vitamin A status was assessed by: clinical indicators (ophthalmic examination), subclinical indicators (questionnaire looking for night blindness, impression cytology with transfer test (ICT), modified relative dose response test (MRDR), and a diet inquiry about vitamin A intakes. Acute malnutrition was assessed by a weight/height measure. RESULTS: In both districts, all the clinical indicators were below the WHO criteria that define a public health problem. Both the ICT test, respectively 19 and 21% of abnormal tests, and the MRDR, 67.3 and 73.1% of tests > or = 0.06, indicated a subclinical vitamin A deficiency as defined by WHO thresholds. During the preceding week only four children in NS had consumed vitamin A-rich food less than seven times. No significant difference between the two districts was found either for clinical or subclinical indicators (p > 0.5). CONCLUSION: Despite a rapid urbanisation, the vitamin A status of the children seemed to be rather homogeneous among the different districts. The population remained vulnerable with peripheral depletion and low hepatic stores of vitamin A. The urban children should be carefully monitored regarding vitamin A status. PMID- 11484805 TI - Rodents and Leptospira transmission risk in Terceira island (Azores). AB - The role of rodents as Leptospira renal carriers in Terceira island was evaluated (1993-1995) through kidney culture and serology [microscopic aglutination test (MAT)] of 94 mice and rats. Fifty-nine animals were positive (n = 41 by serology + culturing; n = 11 serology; n = 7 culturing), presenting a wide distribution in man-made and natural areas. House mice had the highest bacteriological (82.9%) and serological (90.9%) rates, being strictly related to serovar arborea. Black rats were involved in the dispersion of all isolated L. interrogans sensu lato serovars (arborea, copenhageni and icterohaemorrhagiae). Logistic regression analysis and non-metric multi-dimensional scaling, relating Leptospira infection with biological and environmental variables, expressed that adult males Mus domesticus, sexually active and living in humid biotopes, mainly above 500 m, are the most likely reservoirs. This study emphasizes the role of house-mice in the epidemiology of leptospirosis in Terceira and the need of reducing the risk of Leptospira transmission through integrated control programmes, primarily focusing on adult house-mice in peri-domestic environments, before the breeding season. PMID- 11484806 TI - The prevalence of hepatitis C virus infection in patients with lichen planus in Gaziantep region of Turkey. AB - The purpose of this case-control study was to investigate the association between lichen planus (LP) and hepatitis C virus (HCV) infection in Gaziantep region of Turkey. Seventy-three patients with LP and a control group of patients (n: 73) with a dermatological disorder other than LP were detected for HCV infection using a third generation enzyme-linked immunosorbent assay (ELISA). A serological positivity for HCV was found in five of LP patients (6.84%), whereas it was positive for only one patient of the control group (1.36%). A statistically significant difference was found between LP and control groups (p < 0.05). We conclude that the coexistence of the two diseases is probably more than coincidental. PMID- 11484807 TI - Parental education, body mass index and prevalence of obesity among 14-year-old boys between 1987 and 1997 in Wroclaw, Poland. AB - The main aim of this study was to examine changes in relative weight and prevalence of obesity across a ten-year period among 14-year-old boys according to parental education level. Data from two surveys, carried out in 1987 and 1997, of boys attending the 7th grade of primary schools in Wroclaw were used in the analysis. The heights and weights of 3165 boys aged 14 years selected from cohort of 6969 7th and 8th grade boys from all primary schools of the city Wroclaw were used. The data of the second sample of 14-year-old boys (n = 1014) were obtained from a health examination study carried out in the Silesian Centre for Preventive Medicine, 'DOLMED', in Wroclaw in 1997. All boys attended the 7th grade of 34 randomly selected primary schools from a total of 129 schools in the city of Wroclaw. Social status was assessed on the basis of parental education level scored to four categories: university, secondary school, trade school, and elementary school. The prevalence of overweight and obesity was defined as the percentage of children above the 85th and 95th percentiles of the body mass index (BMI), the means of which were 21.27 and 23.75 kg/m2 respectively. Prevalence of overweight among boys is slightly lower in the 1997 sample, whereas the prevalence of obesity shows the opposite trend and is higher by more than one percent in comparison with the 1987 sample. Similar trends of declining medians and increasing variance are observed in all educational groups. The differences in medians between the two samples within educational groups did not achieve statistical significance for the groups with parents with education at elementary level and fathers with university education. There is a trend toward increasing prevalence of obesity across the decade considered, according to father's education level. With respect to mother's education levels, the most dramatic changes in BMI and obesity occurred in the elementary education group, where the percentage of obese subjects increased more than twofold. A significant increase is also observed in the group with parents attaining university education. PMID- 11484808 TI - Characterization of the smoking habit among high school students in Syria. AB - OBJECTIVES: To establish an accurate estimate of the prevalence of smoking among high school students in Aleppo-Syria and to characterize the smoking habit in this age group. DESIGN: A cross-sectional survey was performed, in which a self administered questionnaire was distributed to 10th, 11th and 12th graders. SUBJECTS: A sample of 1587 students (784 males; mean age 16 years, and 803 females; mean age 15.9 years) was studied. This sample was drawn from 16 schools randomly selected from a total of 73 schools in the city. RESULTS: The prevalence of current smoking was 15.9% for males and 6.6% for females. The prevalence of daily smoking was 2.4% for males and 0.5% for females. Almost all smoking categories increased with age and grade. Forty seven percent of male daily smokers were heavy smokers. Peer influence was evident in all stages of the smoking process, especially for males. Parental smoking was also an important associated factor. The combination of parental and siblings smoking was the strongest predictor of the smoking status of the surveyed (OR: 4.4; 95% CI: 2.7 7). Thirty eight percent of male smokers and 47.2% of female smokers smoke out of boredom. Fifty one percent of male smokers were introduced to smoking by a friend. Forty percent of male smokers buy their cigarettes, while 56.6% of female smokers obtain their cigarettes from home. Logistic regression analyses showed that parental and siblings smoking were the most important predictors of ever trying to smoke or current smoking among male and female students. CONCLUSIONS: Our study shows that smoking is more frequent in male than female students. Parental and sibling smoking are the most important factors associated with smoking in this population, and peer influence is important particularly in the smoking-initiation stage. Pattern of smoking and associated factors differed substantially between males and females in this study. PMID- 11484809 TI - Impact on immunization of seasonal cycle of chickenpox. AB - The objective of this work was to study the effect of incorporating a seasonal term in a compartmental age-structured theoretical transmission model, on the performance of vaccination strategies for chickenpox. The epidemiologic parameters are taken from data or are based on a review of literature. We used a realistic age-structure model (RAS): It is a model that incorporate both age and seasonal structure. The seasonality was introduced in the model with the addition of sinusoidal forcing term. This seasonal term varies between 0 and 1. From a susceptible/exposed/infectious/removed (SEIR) model with or without age structure, the pattern of time series is dramatically modified after introducing a seasonal term. This dynamic ends to the transition to chaos. Regarding the evaluation of vaccination strategies, the results of the model without seasonality may be disturbed when the seasonality is introduced, especially during the first 30 years of simulations. PMID- 11484810 TI - Prevalence of hepatitis C infection in Malawi and lack of association with sexually transmitted diseases. AB - To investigate the role of sexual transmission for Hepatitis C virus (HCV) we studied its prevalence in sub-Saharan Africa where sexually transmitted diseases (STDs) are prevalent. Overall, HCV prevalence was 3.9% and similar in 206 STD patients, 127 dermatology patients, and 100 blood donors. No association with HIV or syphilis was observed. Despite high prevalence of STDs, sexual transmission does not appear to significantly contribute to HCV transmission in Malawi. PMID- 11484811 TI - Is volcanic soil a cofactor for classic Kaposi's sarcoma? PMID- 11484812 TI - Marked increase of neuronal prion protein immunoreactivity in Alzheimer's disease and human prion diseases. AB - In neurodegenerative disorders including Alzheimer's disease (AD), free radical damage to lipids, carbohydrates, proteins and DNA has been demonstrated to play a key pathogenetic role. In vitro studies have suggested a function of the cellular prion protein (PrPc) in the defense against oxidative stress. Therefore, we investigated the distribution of PrPc immunoreactivity in hippocampus (sectors CA4-CA1), subiculum (Sub), entorhinal (EC), and temporal cortex (TC) in sections from AD, human transmissible spongiform encephalopathy (TSE) and control brains. Compared to control cases, AD brains revealed an increase in the proportion of PrPc-immunoreactive neurons, which was statistically significant in CA2, Sub, and TC. In TSEs, a statistically significant increase of PrPc-immunoreactive neurons was observed in CA2, CA1, Sub, EC, and TC. In conclusion, our data show a striking up-regulation of PrPc in neurodegeneration and provide additional support for the concept that PrPc may be involved in the defense against oxidative stress. PMID- 11484813 TI - Immunomorphological sequelae of severe brain injury induced by fluid-percussion in juvenile pigs--effects of mild hypothermia. AB - Severe traumatic brain injury (TBI) often leads to a bad outcome with considerable neurological deficits. Secondary brain injuries due to a rise of intracranial pressure (ICP) and global hypoxia-ischemia are critical and may be reduced in extent by mild hypothermia. A porcine animal model was used to study the effect of severe TBI, induced by fluid percussion (FP; 3.5+/-0.3 atm) in combination with a secondary insult, i.e., temporary blood loss with hypovolemic hypotension. Six-week-old juvenile pigs were subjected to this kind of severe TBI; one group was then submitted to moderate hypothermia at 32 degrees C for 6 h, starting 1 h after brain injury. Animals were killed after 24 h. TBI and hypothermia-associated alterations in the brains were investigated by immunohistochemistry with antibodies against microtubule-associated protein 2 (MAP-2) and beta-amyloid precursor protein (betaAPP). In addition, DNA fragmentation was investigated by the terminal deoxynucleotidyltransferase mediated dUTP-biotin nick end labeling (TUNEL) method. Seven of the 13 normothermic TBI animals developed a secondary increase in ICP (TBI-NT-ICP) after an interval of several hours. None of the animals in the hypothermic trauma (TBI HT) group exhibited a secondary ICP increase, indicating a protective effect of the treatment. TBI-HT animals showed significantly higher levels of MAP-2 immunoreactivity, lower levels of betaAPP immunoreactivity and less DNA fragmentation than the TBI-NT-ICP animals. Differences between the TBI-HT group and normothermic animals without an ICP increase (TBI-NT) were less marked. A considerable decrease in MAP-2 outside the site of TBI-FP administration was seen only in the TBI-NT-ICP animals. MAP-2 immunohistochemistry was thus a reliable marker of diffuse brain damage. Axonal injury was present in all TBI groups, indicating its special significance in neurotrauma. Thus, severe TBI caused by FP, combined with temporary blood loss, consistently produced traumatic axonal injury and focal brain damage. Mild hypothermia was able to prevent a secondary increase in ICP and its sequelae of diffuse hypoxic-ischemic brain injury. However, hypothermia did not afford protection from traumatic axonal injury. PMID- 11484814 TI - Ultrastructural findings at the satellite cell-myofiber border in normal and diseased human muscle biopsy specimens. AB - Satellite cells (SC) are mononuclear myoblasts located between the plasma membrane and the basement membrane of a myofiber. In normal adult muscle, SC are quiescent in the G0 phase of the cell cycle. The contact of the SC with the myofiber plasma membrane imposes a mitotic inhibition on the SC. Sarcolemmal molecules which might explain this membrane-imparted mitotic inhibition have not yet been identified. In this study we examined the border of the SC and the adjacent myofiber electron microscopically, assessing the number of SC showing encroachments of basement membrane (BM) material, secretion of cellular degradation products into the intercellular space, and caveolae. We studied normal and diseased muscle including Duchenne muscular dystrophy, Becker muscular dystrophy, idiopathic inflammatory myopathies, and neurogenic atrophies. Caveolae were present in SC from normal muscle, but they were more abundant in SC from diseased muscle, and they significantly prevailed at the outer surface of SC in all of the diseased muscle groups. Encroachments of BM material was only present in SC from diseased muscle, and mostly so in neurogenic atrophies. Secretion of cellular degradation products into the intercellular cleft occurred in normal and diseased muscle. We conclude that degradation products in the intercellular cleft do not disturb SC adhesion and that there is a neural influence on SC adhesion. The significance of the abundance of caveolae at the outer surface of the SC when compared with the inner surface in diseased muscle remains at present unknown. PMID- 11484815 TI - CD8+ phagocytes in focal ischemia of the rat brain: predominant origin from hematogenous macrophages and targeting to areas of pannecrosis. AB - We have recently described a novel population of CD8+ phagocytes that are strongly recruited to focal ischemic lesions of the rat brain but absent from axotomized central fiber tracts. To assess the relative contribution of infiltrating macrophages and resident microglia to the CD8+ phagocyte response, we selectively depleted peripheral macrophages by systemic administration of dichloromethylene diphosphonate-filled liposomes prior to the induction of permanent ischemia by photothrombosis of cortical microvessels. Macrophage depletion led to a dramatic reduction but not complete abolishment of CD8+ cells in the ensuing infarcts. Systemic administration of monoclonal antibody Ox-8 eliminated CD8+ cells from peripheral lymphoid organs but had no effect on CD8+ phagocytes in the ischemic brain lesions. To further characterize the lesion conditions inducing the recruitment of CD8+ phagocytes, we induced mild focal ischemia by transient occlusion of the middle cerebral artery that leads to a core infarction with ischemic pannecrosis surrounded by areas with selective neuronal cell death. Recruitment of CD8+ phagocytes was restricted to areas of ischemic pannecrosis. In areas undergoing selective neuronal loss microglia up regulated complement receptor-3, exhibited ED1 immunoreactivity (indicating phagocytic activity), and to some extent expressed CD4, but not CD8 antigens. In conclusion our present study shows that CD8+ phagocytes in focal brain ischemia are predominantly derived from hematogenous macrophages and selectively target to areas of ischemic pannecrosis. PMID- 11484816 TI - Expression of connexin 43 and connexin 32 gap-junction proteins in epilepsy associated brain tumors and in the perilesional epileptic cortex. AB - The expression of the gap-junction proteins connexin (CX) 43 and 32 was evaluated in surgical specimens of brain tumors and perilesional cortex from patients with chronic medically intractable epilepsy. In human normal brain CX32 was expressed in neurons and oligodendrocytes. CX32 immunoreactivity (IR) was observed in the neuronal component of glioneuronal tumors and in all oligodendrogliomas, 50% of which showed strong labeling, independent of the grade of differentiation. CX43, normally expressed in astrocytes, was also detected in most of the human astrocytomas and in the astroglial component of glioneuronal tumors. Whereas most of the low-grade gliomas (>60%) showed strong membranous staining, most high grade astrocytomas exhibited a reduction of the typical plasma membrane CX43-IR and intracytoplasmic localization. Immunoblot analysis showed different CX43 isoforms in control cortex and in low-grade gliomas. However, only one single isoform (corresponding to the non-phosphorylated form of CX43) appeared to be present in most high-grade gliomas. Increased expression of CX43 protein was present in reactive astrocytes in the epileptic cortex surrounding low-grade tumors as compared to control cortex, indicating the existence of a regulatory pathway involving CX43 in the reorganization of the astrocytic syncytium in regions undergoing reactive gliosis. The high expression of connexin proteins in low-grade tumors and in the peritumoral reactive astrocytes suggests that they could contribute to tumor-related seizures. PMID- 11484818 TI - Development of muscle pathology in canine X-linked muscular dystrophy. II. Quantitative characterization of histopathological progression during postnatal skeletal muscle development. AB - We have characterized the time course of muscle pathology development during the postnatal maturation of quadriceps and tibialis anterior muscle in dystrophic golden retriever dogs. We determined the percentages of degenerating, regenerating, calcium-positive, hypercontracted, albumin-positive, and C3 complement fraction-positive muscle fibers and the extent of connective tissue proliferation in animals from neonate to adult. Necrotic fibers increased from days 2 to 30, decreased at 60 days (to 0.8%) and increased in older animals to a stable level of around 2%. Hypercontracted fibers peaked at 15 days (19.1%) and declined to 3.7% in adults. Regenerating fibers were numerous at 15 and 30 days (10%), declined at 60 days to 4.7% and declined further in adults. Calcium- and albumin-positive fibers peaked at 30 days (6.5% and 13.8%, respectively) and then declined to around 3% and 5%, respectively, in older dogs. In dystrophic dogs, the extent of fibrosis was significantly greater on 15 days than in controls, but did not then increase with age. In carriers, calcium- and albumin-positive fibers always expressed dystrophin abnormally. Muscle damage occurs before completion of muscle maturation in dystrophic dogs. While necrosis and hypercontraction remain stable in adults, fiber regeneration declines to very low levels. In contrast to Duchenne muscular dystrophy, muscle fibrosis in the muscle studied does not increase with age. PMID- 11484817 TI - Kainate-induced epilepsy alters protein expression of AMPA receptor subunits GluR1, GluR2 and AMPA receptor binding protein in the rat hippocampus. AB - Kainic acid induces seizures with consecutive degeneration of highly vulnerable hippocampal CA3 neurons in adult rats. An abnormal influx of calcium through newly synthesized alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid (AMPA) receptors lacking the GluR2 subunit, which normally renders AMPA receptors calcium impermeable, is thought to play a pivotal role for postictal neuronal death (GluR2 hypothesis). Using a specific GluR2 antiserum, postictal hippocampal GluR2 protein expression was investigated and compared to GluR1 between 6 and 96 h after seizure induction. In addition, postictal protein expression of a recently cloned AMPA receptor binding protein (ABP), which anchors AMPA receptors in the plasma membrane was also analyzed, to address the question of whether its protein expression is associated with neuronal death or survival. At 6 h after seizure induction, GluR2 immunoreactivity (IR) in CA3 was more markedly reduced compared to GluR1, but at 24 h GluR2 IR reattained control levels. More importantly, GluR2 IR was also markedly, but transiently decreased between 6 and 48 h in hippocampal CA1 neurons, but no significant cell loss was observed. These findings modify the GluR2 hypothesis in so far as only a subset of, but not all, hippocampal CA1 and CA3 pyramidal neurons may die due to reduced GluR2 levels with consecutive calcium overload through calcium-permeable AMPA receptors. ABP was induced postictally in presumed CA2 and a subpopulation of CA3 neurons and seems not to be involved in mechanisms of delayed neuronal death. PMID- 11484819 TI - Mutational analysis of INI1 in sporadic human brain tumors. AB - The INI1/SMARCB1/hSNF5 gene on chromosome 22 is frequently mutated in rhabdoid tumors. An association of INI1 mutations with allelic losses on chromosome 22 supports a classical tumor suppressor mechanism. Several brain tumor entities including astrocytomas, glioblastomas and ependymomas are characterized by allelic losses on chromosome 22. In the present study we examined a series of 200 brain tumors by Single-strand conformation polymorphism analysis and direct sequencing for point mutations in INI1. In addition, all tumors were analyzed for homozygous deletions spanning both exons 3 and 8 of INI1. No mutations or homozygous deletions were detected in astrocytomas, glioblastomas, oligodendroglial tumors, neurinomas or medulloblastomas. However, a point mutation could be identified in the single case of plexus carcinoma. Our data suggest that INI1 mutations are involved in the pathogenesis of plexus carcinoma; however, INI1 alterations are not a frequent event in the majority of brain tumor entities. PMID- 11484820 TI - Nuclear shrinkage in live mouse hippocampal slices. AB - Brain slices are used extensively for biochemical, electrophysiological and molecular investigations. However, only the time frame for electrophysiological and biochemical investigations has as yet been defined. The goal of the present study was to investigate the time course of nuclear structure in live brain slices. Hippocampal slices (300 microm) were prepared from male CD1 mice (25-30 g), stained with Hoechst 33342 (10 microM), calcein-AM (2 microM) and ethidium homodimer (4 microM), and imaged with single- and dual-photon microscopy. The volume of CA1 pyramidal cell nuclei decreased from 759+/-229 microm3 in 40-50 microm depth 25 min after preparation to 453+/-169 microm3 (P<0.001) after 60 min, 315+/-112 microm3 (P<0.001) after 120 min and 128+/-71 microm3 (P<0.001) after 8 h. Similar results were obtained on a prolonged time scale in 70-80 microm depth and with an accelerated time scale in 20-30 microm depth. Live-dead staining showed that cell damage is progressing from the surface to deeper layers of the slices in a time-dependent fashion. We conclude that nuclei of CA1 hippocampal pyramidal cells show a time- and depth-dependent shrinkage converging 8 h after slice preparation to a volume of 90-130 microm; in any depth between 20 and 80 microm. The nucleus in the superficial 80 microm of each side appears dysfunctional even at times suitable for electrophysiological and biochemical experimentation in hippocampal slices. Molecular analysis of cell regulation in brain slices may, therefore, be time-dependently distorted by progressing cell death in at least half of the tissue under investigation. PMID- 11484821 TI - Morphological alterations in the inner ear of the arylsulfatase A-deficient mouse. AB - Metachromatic leukodystrophy of humans is an inherited sulfatide lipidosis due to deficiency of arylsulfatase A (ASA). As an animal model, ASA(-/-) mice have been generated. A previous study showed that the mice lose most of their spiral (acoustic) ganglion cells and develop deafness by the end of the first year of life. The present report describes the sulfatide histochemistry and ultrastructure of the inner ears of ASA(-/-) mice at 0.5-26 months of age. Lysosomal accumulation of sulfatides was observed in various cell types such as Schwann cells that maintain the myelin sheaths around the spiral and vestibular ganglion cells, periaxonal Schwann cells, macrophages, and spiral and vestibular ganglion cell perikarya. In the spiral ganglion, the only surviving neurons were those which are primarily non-myelinated (type 2 cells). However, the myelinated spiral neurons and their processes were rarely encountered within the process of dying, suggesting that this was a rather rapid process. Since the myelin sheaths around dying perikarya and axons appeared structurally normal, the primary cause of the neuronal cell death seems to reside in the neuron. In contrast to the spiral ganglion, the vestibular ganglion as a whole survived throughout the period of observation. The organ of Corti and the vestibular apparatus appeared preserved at the light microscopic level, despite massive sulfatide storage in the vestibular hair cells. PMID- 11484822 TI - A mouse model of Streptococcus pneumoniae meningitis mimicking several features of human disease. AB - The course of bacterial titers, meningeal inflammation, behavioral abnormalities, and neuronal damage was studied in a mouse model of Streptococcus pneumoniae meningitis. At 24 h after injection of 10(4) colony-forming units (CFU) S. pneumoniae into the right forebrain, infected mice became severely lethargic. Bacterial titers in cerebrospinal fluid and cerebellum rose to 10(9) CFU/ml, with strong granulocyte invasion into the meninges and neuronal necroses in the neocortex, striatum and hippocampal formation. Meningeal inflammation and neuronal damage in intercellular cell adhesion molecule-1- and macrophage colony stimulating factor-deficient mice was similar to that in wild-type littermates. Untreated, the infection was fatal. Wild-type mice treated earlier than 24 h after infection with ceftriaxone (2 mg every 12 h for 3 days) survived without apparent behavioral abnormalities. Delay of treatment beyond 30 h led to the death of more than 50% of the infected mice. This mouse model is suitable for therapeutic studies and for the investigation of inflammation in knockout mice. The neuronal damage resembles morphological abnormalities observed in humans. PMID- 11484823 TI - Tubular aggregates observed in spindle muscle fiber of horse lumbrical muscle. AB - Tubular aggregates (TAs) originate from the sarcoplasmic reticulum (SR) and form polymorphic double (or single) -walled structures in cross section. TAs are involved in various human skeletal muscle disorders including periodic paralysis, congenital myasthenic syndromes, inflammatory myopathies, and malignant hyperthermias. Horse lumbrical muscle (LM) is a slender fusiform muscle that shows varying degrees of regression due to its limited activity in the limb. Double-walled TAs were found in degenerating spindle fibers and with a range of 80-116 nm (average 92 nm, n=135) for outer layer and 50-78 nm (average 59 nm, n=135) for the inner layer. TAs exhibit degradation of myofibrillar proteins, disruption of mitochondria with cristae lost, glycogen accumulation, electron dense metabolic products, blebbing appearance of sarcolemma, and presence of various vacuoles. LM fibers also show a similarly degenerative state. The disassembly of the SR structure probably produces a large accumulation of SR proteins which remain as molecules without being further degraded and which could aggregate to form the orderly structure of TAs. We believe that TA formation may be an adaptation to store unbalanced extra proteins by forming ordered aggregates in degeneration caused by stress in cells. PMID- 11484824 TI - Tau isoform profile and phosphorylation state in dementia pugilistica recapitulate Alzheimer's disease. AB - Insights into mechanisms of familial Alzheimer's disease (AD) caused by genetic mutations have emerged rapidly compared to sporadic AD. Indeed, despite identification of several sporadic AD risk factors, it remains enigmatic how or why they predispose to neurodegenerative disease. For example, traumatic brain injury (TBI) predisposes to AD, and recurrent TBI in career boxers may cause a progressive memory disorder associated with AD-like brain pathology known as dementia pugilistica (DP). Although the reasons for this are unknown, repeated TBI may cause DP by mechanisms similar to those involved in AD. To investigate this possibility, we compared the molecular profile of tau pathologies in DP with those in AD and showed that the same tau epitopes map to filamentous tau inclusions in AD and DP brains, while the abnormal tau proteins isolated from DP brains are indistinguishable from the six abnormally phosphorylated brain tau isoforms in AD brains. Thus, these data suggest that recurrent TBI may cause DP by activating pathological mechanisms similar to those that cause brain degeneration due to accumulations of filamentous tau lesions in AD, and similar, albeit attenuated, activation of these processes by a single TBI may increase susceptibility to sporadic AD decades after the event. PMID- 11484825 TI - Rosetted glioneuronal tumor: a case with proliferating neuronal nodules. AB - Glioneuronal tumors with neuropil-like islands (rosetted glioneuronal tumors) were recently reported as a novel brain tumor entity with characteristic clinicopathological features (Am J Surg Pathol 23: 502, 1999). Here we describe the clinical, histological and genetic features of another case arising in the parietal lobe of a 43-year-old man suffering from focal motor epilepsy. Histologically, nodules of small neuronal tumor cells immunoreactive for synaptophysin and NeuN were embedded within a diffuse astrocytoma. Remarkably, highest proliferative activity was observed within the neuronal nodules. Comparative genomic hybridization revealed a gain of chromosome 7q and a loss on chromosome 9p. PMID- 11484826 TI - Chronic inflammatory demyelinating polyradiculoneuropathy with histologically proven optic neuritis. AB - Although patient series of clinical, electrophysiological, or magnetic resonance imaging evidence for involvement of the central nervous system in patients with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) have been published, the histological proof has never been reported. We present the case of a 46-year-old male patient who developed CIDP in his early 20s and who died of relapsing severe pneumonia. In late stages of the disease the patient presented visual loss and bilateral atrophy of the optic nerve. Neuropathological examination revealed severe peripheral neuropathy consistent with CIDP and central involvement with bilateral optic neuritis. This is the first case reporting CIDP and histologically proven optic neuritis. PMID- 11484827 TI - Effects of double UVB-irradiations with different intervals on the dorsal skin of wistar-derived hypotrichotic WBN/ILA-Ht rats. AB - The effects of double irradiations of UVB (10 kJ/m2) with different intervals (12, 24 and 48 hr) were examined on the dorsal skin of Wistar-derived hypotrichotic WBN/ILA-Ht rats as the first step for long-term repeated irradiation study. The dorsal skin responses to a single UVB-irradiation were similar to those previously reported by our research group (Exp Toxicol Pathol, in press). In the groups which were given the 2nd irradiation at 12 and 24 hr after the 1st one, erythema, epidermal damage and subsequent hyperplasia, intradermal inflammatory cell infiltration and edema developed earlier and were more prominent especially in 12 hr-group compared with a single irradiation. However, the sequence of sunburn cells, the most characteristic epidermal change, was not different from that after a single irradiation. On the other hand, the dorsal skin responses to the 2nd irradiation at 48 hr after the 1st one were almost similar to those observed in the single irradiation-group except for epidermal hyperplasia being more prominent in 48 hr-group. This suggests that the responsibility of the dorsal skin almost recovered at 48 hr after the 1st irradiation. PMID- 11484828 TI - Regression of large tumors expressing a suicide gene. AB - Using the HSV-tk/GCV system, complete remissions of tumors up to a size of 5 mm in diameter have been reported in mice and rats. In order to examine whether larger tumors of clinically relevant size can also be successfully treated, we established hepatic and peritoneal tumors consisting of retrovirally pretransduced tk-positive CC531 colon adenocarcinoma cells in syngenic Wag/Ola rats. In this model, we evaluated whether large tumors respond to therapy under the ideal condition of 100% gene transfer. Within 10 days, GCV treatment led to a marked regression of the adenocarcinomas. Tumors up to a size of 4000 mm3 could be completely eradicated. Passing through several stages of degeneration and cytolytic necrosis, a complete replacement by fibrous tissue was seen. There was no histological evidence for apoptosis and cell mediated immunity. These results suggest that complete regression of tumors of clinically relevant size can be achieved by direct toxic effects of the HSV-tk/GCV system if 100% of the cells are transduced. PMID- 11484829 TI - Immunohistochemical study on hypoxia in spontaneous polycystic liver and kidney disease in rats. AB - Hypoxia-inducible factor (HIF) mediates homeostatic responses to hypoxia and activates transcription of hypoxia-inducible genes including vascular endothelial growth factor (VEGF). The aim of this study was to examine the expressions of VEGF, HIF-1alpha and HIF-3alpha in spontaneously occurring hepatorenal polycystic lesions in two Sprague-Dawley (Crj:CD) rats. Hepatic multiple cysts were derived from the interlobular and large bile ducts, while renal cysts were from the collecting ducts and distal tubuli. These findings were confirmed by a lectin peanut agglutinin (PNA) histochemistry. In the polycystic liver, VEGF immunoreaction was strongly evident in the cytoplasm of hepatocytes, whereas expression of HIF-3alpha, but not HIF-1alpha, was found in a few nuclei of hepatocytes. In the polycystic kidney, VEGF immunoreaction was increased in the cytoplasm of collecting ducts and distal tubuli, whereas nuclear expression of HIF-1alpha and HIF-3alpha was evident in the proximal tubuli and thin loop of Henle, respectively. The results suggest that hypoxia-related molecules may be induced by cystic alterations in a heterogeneous appearance. PMID- 11484830 TI - Effects of atropine, trimedoxime and methylprednisolone on the development of organophosphate-induced delayed polyneuropathy in the hen. AB - In this study we have examined the effects of atropine, trimedoxime (TMB-4) and methylprednisolone (MP) on the development of organophosphate-induced delayed polyneuropathy (OPIDP) in the hen. The birds were treated with standard neuropathic dose of diisopropylfluorophosphate (DFP) (1.1 mg/kg, sc), which produced OPIDP that could be graded as 5 on the 8-point scale, and the development of OPIDP was observed for the next 22 days. The results obtained have shown that atropine (20 mg/kg, ip), TMB-4 (15 mg/kg, im) and MP (2 or 10 mg/kg, ip) either alone or in different combinations are able to improve the condition of the birds. The most potent effect was obtained with atropine, TMB-4 (given 20 min before DFP) and MP (2 mg/kg, sc, given 20 min before and at 48 hour intrevals after poisoning) since the signs of OPIDP could hardly be seen (grade 1 at the 8 point scale). When TMB-4 and MP were given 15 or 40 min after DFP the protective/therapeutic effects of these drugs appeared to be diminished since walking disorders were more serious and graded as 2 or 4, respectively. The possible mechanisms of the action of the drugs in respect to OPIDP are discussed. In conclusion, the results of this study have shown that it is possible to prevent the development of DFP-induced OPIDP in the hen by treatment with atropine, trimedoxime and methylprednisolone when they were given before or soon after DFP. PMID- 11484831 TI - In vitro effects of NSAIDS and paracetamol on oxidative stress-related parameters of human erythrocytes. AB - In vitro effects of widely used nonsteroidal antiinflammatory drugs (NSAIDs) and paracetamol were studied on oxidative stress-related parameters of human red blood cells (RBC). Membrane lipid integrity, activity of erythrocyte antioxidant enzymes; i.e. glutathione S-transferase (GST), selenium dependent-glutathione peroxidase (Se-GPx), and catalase (CAT), and hemolytic/stabilizing action of the drugs on erythrocyte membrane were assessed. Diclofenac, indomethacin and paracetamol at the therapeutic and higher concentrations, and dipyrone at the high concentration exerted a statistically significant inhibition on H2O2 forced erythrocytic membrane lipid peroxidation (EMLP). Increased hemolysis was observed by Na-salicylate, naproxen and ketorolac at therapeutic and higher concentrations, and by diclofenac and tiaprofenic acid at high concentrations, while the others seemed to stabilize the membrane at the same conditions. Na salicylate inhibited GST activity at the therapeutic dose, however activated the same enzyme at high concentrations. Naproxen, tiaprofenic acid and piroxicam caused a decrease in GST activity at therapeutic doses. Paracetamol caused an activation at a high dose. Tiaprofenic acid, ketorolac, naproxen and piroxicam caused a significant Se-GPx inhibition. Erythrocyte CAT activity was increased by Na-salicylate, acemetacin, and tenoxicam at the therapeutic, and by dipyrone at the high concentration. Our results suggest that NSAIDs and paracetamol may be involved in oxidative/antioxidative processes of human erythrocytes. Also, the in vitro EMLP method can be considered as a simple test for evaluating possible antioxidant potency of chemicals. PMID- 11484832 TI - A method for in vitro analysis of the biological activity of complex mixtures such as sidestream cigarette smoke. AB - Studies of the cytotoxicity of air contaminants such as gaseous or particulate compounds and complex mixtures have traditionally used in animal experiments because of the difficulties in exposing cell cultures directly to these substances. New cultivation and exposure techniques enhance the efficiency of in vitro methods, as demonstrated by a new system called CULTEX* which uses a transwell membrane technique for direct exposure of complex mixtures like sidestream cigarette smoke at the air/liquid interface. The factors influencing the susceptibility of human bronchial epithelial cells (e.g. gas flow rate or duration of exposure) were studied and the cells were finally exposed for one hour to clean air or different concentrations of sidestream smoke. The biological parameters estimated were number of cells, metabolic activity and glutathione concentration. After exposure of the cells to sidestream cigarette smoke, dose dependent effects were measured. Thus, the introduction of these cultivation and exposure techniques offers new testing strategies for the toxicological evaluation of a broad range of airborne and inhalable compounds. PMID- 11484833 TI - Hypertonic-hyperoncotic solutions decrease cardiac troponin I concentrations in peripheral blood in a porcine ischemia-reperfusion model. AB - In this study we addressed the question of whether the measurement of cardiac Troponin I (cTnI) is able to reflect beneficial effects of hypertonic hyperoncotic solutions after transient cardiac arrest. Ten pigs were anaesthetized and cardiac arrest was induced by electric fibrillation. After 5 minutes of global ischemia, cardiac arrest was reversed by electric defibrillation. Upon return of spontaneous circulation 5 animals received hypertonic-hyperoncotic solutions (10% Hydroxyethylstarch 200/0.5 and 7.2% NaCl). The other animals received equivalent volumes of physiological saline. We observed that cTnI serum levels of animals treated with hypertonic-hyperoncotic solutions were significantly lower than those treated with saline. We conclude that hypertonic-hyperoncotic solutions may have cardioprotective effects. PMID- 11484834 TI - Age-related incidence of spontaneous non-neoplastic lesions in a colony of Han:AURA hamsters. AB - In toxicologic testing or experimental studies using animals, an adequate knowledge of spontaneously occurring lesions is required. 144 male and 184 female untreated Syrian golden hamsters (strain Han:AURA) were kept for life under standard laboratory conditions and an investigation of non-neoplastic lesions in relationship to the lifespan was performed. The average lifespan of the males was 106 weeks and that of the female hamsters 97 weeks. While cartilage degeneration of the sternum and fatty degeneration of the femoral bone marrow occurred already in the first half of life with high incidence, the majority of lesions were observed in the second half. The most frequent non-neoplastic changes in various organs were fatty change, calcification, cystic change, hyperplasia and amyloidosis. Such spontaneous lesions were discussed in connection with the same alterations which can also be induced by chemical or hormonal agents. PMID- 11484835 TI - M-CSF transgenic mice: role of M-CSF in infection and autoimmunity. AB - In this study, transgenic CD2F1 mouse lines (C-1.1-C-1.11) bearing a transgene encoding the murine growth factor M-CSF under the control of the liver specific alpha-1-antitrypsin gene promoter were generated. Transgenic C-1.4 mice showed elevated expression of transgene-encoded M-CSF in the liver and displayed a 2-3 fold increase of M-CSF plasma levels and of macrophage numbers in the liver as compared with non-transgenic littermates. M-CSF transgenic mice showed increased resistance against sublethal i.v. infections with Listeria monocytogenes as compared with infected non-transgenic mice. To investigate the influence of M-CSF in murine systemic lupus erythematosus (SLE), the M-CSF transgenic mouse line C 1.4 was bred into the genetic background of SLE-prone MRL+/+ mice. The resulting C-1.4/MRL transgenic mice bearing increased endogenous M-CSF levels showed consistently lower levels of anti-ss-DNA autoantibodies as compared with non transgenic MRL+/+ mice. The life span of the C- 1.4/MRL transgenic mice and the severity of the disease in these mice remained unchanged as compared with their non-transgenic littermates. It is concluded that in addition to M-CSF further factors must be involved in the acceleration of the autoimmune disease in SLE prone MRL/lpr mice. PMID- 11484836 TI - Protective effect of mannitol, glucose-fructose-sucrose-maltose mixture, and natural honey hyperosmolar solutions against ethanol-induced gastric mucosal damage in rats. AB - BACKGROUND: We have previously shown that natural honey is able to protect the rat stomach against acute ethanol- and indomethacin-induced lesions. The present investigations were undertaken to examine the role of intraluminal osmolality in this protective effect. METHODS: Mannitol, glucose-fructose-sucrose-maltose mixture (GFSM) and natural honey (300, 600, 1800 mOsmol/kg water) were given orally to rats 30 min before administration of 70% ethanol for a further 15-min period. Lesions area of the excised stomachs were evaluated. Pylorus-ligated stomachs were filled with mannitol, GFSM mixture and honey (1800 mOsmol/kg water) to test the effect of the hyperosmolar solutions on gastric fluid content and acid secretion. The rate of gastric emptying of the three test solutions (1800 mOsmol/kg) was measured by the phenol red method. RESULTS: Intragastric administration of mannitol, GFSM mixture or honey prevented the formation of mucosal lesions in an osmolality-dependent manner. Using the pylorus-ligated stomach model, the test solutions led to a net increase of luminal fluid volume without affecting acid content. Hyperosmolar solutions presented a delayed gastric emptying if compared to a nonnutrient solution made of carboxymethyl cellulose. CONCLUSIONS: The observed results suggest that hyperosmolar solutions can prevent the formation of hemorrhagic lesions by luminal dilution of the necrotising agent and acid, an effect which may be potentiated by a lowered gastric emptying rate. PMID- 11484837 TI - Delayed-type hypersensitivity and humoral immunity modulation by dietary lipids in a murine model of pulmonary tumorigenesis induced by urethan. AB - Mice fed on semisynthetic formulas containing 15% of corn oil (CO), cod fish liver oil (FO), oleic acid (O) or a mixture of 46% of palmitic and 50% of stearic acids (PS) were treated with urethan during 18 weeks for lung tumor induction. Delayed-type hypersensitivity (DTH) assay, hemagglutination assay and the amount of lung nodes (alveolar adenocarcinomas) were recorded. Results showed significantly greater DTH in CO and FO with respect to O and PS feeding mice; the two last ones induced an essential fatty acid (EFA) deficiency (EFAD). In the O lot there was a non-significant diminution of the humoral response. EFAD animals exhibited a tendency to increase number of lung nodes in relation to CO and FO lots. Splenomegalia was recorded in FO lot. Confront between spleen weight and DTH showed a 72% correlation, suggesting an increase in cellular immunity as increasing unsaturation. It may be concluded that in this suitable model of tumorigenesis the manipulation of dietary lipids may be a strategy to modify the immune system response. PMID- 11484838 TI - Local proliferation contributes to lymphocyte numbers in normal lungs. AB - Lymphocytes in the bronchoalveolar lavage fluid (BAL) are increased in many lung diseases, which might be an indicator for protective reactions or pathology. Higher lymphocyte numbers at distinct organ sites may be due to a number of reasons such as increased entry, increased proliferation, reduced apoptosis or reduced exit. It is not known whether lymphocyte numbers are influenced by local proliferation in the healthy lung. Therefore, the proliferation of lymphocytes was studied in vivo in different lung compartments of healthy rats: the marginal vascular pool, the interstitial pool and the bronchoalveolar pool. Bromodeoxyuridine (BrdU) was used as an S-phase proliferation marker. The cells were obtained 1 h and 24 h after i.v. injection of BrdU. The labeled cells and their phenotypes were determined by immunocytochemistry, since it was not possible to use flow cytometry because of the low numbers. In the lung compartments 0.7-1.5% of all nucleated cells were found to be BrdU+, whereas 24 h later this increased to 3.2-5.7%. The frequency of BrdU+ T cells was significantly higher in the lung compartments compared to the blood, with the maximum in the marginal vascular pool. Local proliferation of lymphocytes involved mainly CD8+ T cells. Thus, local proliferation plays a role in the number and composition of lymphocytes in the healthy normal lung. PMID- 11484839 TI - The usefulness of antioxidative enzymes for the estimation of synthetic effects of PGE1 analogue. AB - The aim of our research was estimation of the activity of superoxide dysmutase (SOD) and glutathione peroxidase (GPx) in rabbits' blood after administration of single dose of carbon tetrachloride (CCl4) and synthetic analogue PGE1 - Cytotec. The research was carried out on 56 Chinchilla rabbits. By means of stomach tube rabbits were administered respectively: CCl4, Cytotec, CCl4 with Cytotec, indomethacin, Cytotec with indomethacin and CCl4 with Cytotec and indomethacin simultneously. In comparison with control group almost double increase of SOD activity was stated at rabbits which were administered CCl4 and CCl4 with indomethacin. At rabbits which were administered CCl4 with Cytotec and CCl4 with Cytotec and with indomethacin, the SOD activity increased by only 50% in comparison with control group. At rabbits in whose blood the SOD activity increased, the activity of GPx decreased on average by respectively 40% and 15%. The stated changes of SOD and GPx activity in rabbits' blood which were administered prostaglandin apart from CCl4 indicates prostaglandin's indirect interference into hepatocytes antioxidative system and thus its hepatoprotective activity. PMID- 11484840 TI - Pharmacological modification of endogenous antioxidant enzymes by ursolic acid on tetrachloride-induced liver damage in rats and primary cultures of rat hepatocytes. AB - The purpose of this study was to investigate possible protective effects of ursolic acid against CCl4-induced alterations of antioxidant defence enzymes in vivo as well as its effects against CCl4-intoxication in vitro. Pre-treatment of rats with ursolic acid significantly reduced serum levels of glutamate-oxalate transaminase and glutamate-pyruvate-transaminase previously increased by administration of CCl4. Treatment with ursolic acid also significantly reversed the decreased superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase activities and glutathione levels in the liver, as the concentration of reduced glutathione was increased and the content of oxidized glutathione decreased in ursolic acid treated groups. Levels of lipid peroxidation were higher in the CCl4 group but the increase was also reduced after drug treatment (p < 0.01 for 1, 2.5 and 5 mmol/kg). In vitro results indicated that addition to the culture medium of ursolic acid (p < 0.01 for 500 microM) resulted in a reduction of glutamate-oxalate-transaminase, lactate dehydrogenase activities and in a good survival rate for the CCl4-intoxicated hepatocytes. Ursolic acid also ameliorated lipid peroxidation in primary cultured rat hepatocytes exposed to CCl4, as demonstrated by a reduction in malondialdehyde production. Moreover, ursolic acid (50-500 microM) showed radical scavenging properties in terms of hydroxyl formation. The results obtained suggest that ursolic acid treatment can normalize the disturbed antioxidant status of rats intoxicated with CCl4 by maintaining the levels of glutathione and by inhibiting the production of malondialdehyde due to its radical scavenging properties. PMID- 11484841 TI - Effects of 28-day mechanical and chewing stress on content of bound and diffusible ions in muscles of mastication. AB - Type I and type II muscle fibres have different ion concentrations. Muscles adapt to chronic stress by changing of fibre types and remodelling of the myosin heavy chains in the muscle fibres. The present investigation on ionic change during muscular contraction was carried out on 10-week-old pigs (6 treated animals, 6 controls) over a 28-day period. Six pigs received acrylic build-ups to induce mechanical advancement of the lower jaw and chronic chewing stress. Muscle tissue was taken from the masseter (M1, M2, M3), temporal (TP1, TP2), medial pterygoid (PM) and geniohyoid (GH) muscles by a standardized method. Eighty-four muscle samples were used for histological fibre differentiation with mATPase. Energy dispersive X-ray microanalysis of muscles was carried out in an environmental scanning electron microscope. Endurance stress in the stressed muscles was seen as an increase of type I fibres (P < 0.001). This histological change and ionic alterations were measured in the anterior region of the masseter (M1 and M2) and in the posterior region of the temporal muscle (TP2). Smaller changes were found in the medial pterygoid muscle. We measured in this muscles increases in potassium, sulphur, chloride (P < 0.05) and even larger increases in phosphate (up to 1.5 mmol/g to 2.3 mmol/g, P < 0.001) and sodium (3-fold, P < 0.001). The results reveal the effects of chronic stress on muscle fibres and ion concentration in the muscle. Chronic stress resulted in an increase of type I fibres and increased ion concentration in the same muscle region. These are considered to be indicators of more efficient contraction. The changes in ion concentration are an important factor in muscle contraction. PMID- 11484842 TI - Species- and sex-specific renal cytotoxicity of ochratoxin A and B in vitro. AB - Four different cell models were chosen for comparison of OTA and OTB toxicity: primary porcine (PKC), rat (RPTC) and human renal proximal epithelial cells (HKC) from both sexes and a porcine renal cell line: LLC-PK1. Culture conditions were tested and optimized for each respective cell type (species/sex and origin). All cell types were characterized for epithelial origin and growth patterns and following optimization of dosing strategies and assay procedures, a strict study design was implemented to avoid systemic variations. Due to possible sensitivity differences, three simple endpoints were chosen to provide basic data for interspecies comparison: neutral red uptake, MTT reduction and cell number. Of the endpoints tested neutral red appeared the most sensitive, although all three parameters yielded comparable EC50's. Sex-differences were observed between male and female HKC cells following 96 h exposure to OTA, with HKC(m) being more sensitive than HKC(f). No sex-difference was observed in PKC cells, however, the PKC were approximately 3 and 10 times more sensitive than HKC(m) and HKC(f), respectively, to OTA and OTB. Interestingly, the CI95 of the EC50 values obtained for OTA (15.5-16.5 microM) and OTB (17.0-2 1.0 microM) were comparable in the PKC cells. In contrast, OTB had lower cytotoxicity than OTA in HKC and LLC-PK1 (approx. 2-fold) and no effects in RPTC. Overall, HKC(m) were nearly as sensitive as PKC towards OTA, followed by RPTC, LLC-PK1 and HKC(f), thus suggesting a sex specific sensitivity in humans towards OTA induced cytotoxicity. PMID- 11484843 TI - Effect of bile acids on the proliferative activity and apoptosis of rat hepatocytes. AB - Bile acids are known to have damaging as well as protective effects on liver cells. A likely candidate for bile acid-mediated hepatocellular injury during cholestasis is glycochenodeoxycholic acid (GCDCA), a hydrophobic bile acid with a direct cytotoxic effect on hepatocytes. In contrast, ursodeoxycholic acid was shown to exhibit protective effects. Our aim was to determine the effect of GCDCA on proliferation, synthesis and secretion of proteins and death processes in cultured rat hepatocytes. Furthermore, it should be studied whether the hydrophilic bile acid tauroursodeoxycholic acid (TUDCA) might be able to protect cells from the damaging effect of GCDCA. Our results demonstrate that GCDCA decreased dose-dependently hepatocellular proliferation, synthesis and secretion of newly synthesized proteins and, at low concentration, induced apoptosis or, at high doses, cytolysis of cultured hepatocytes. TUDCA did not exert cytotoxic effects on the isolated hepatocytes at a wide range of concentrations. However, TUDCA coincubated with GCDCA protected the cells from the damaging effect of GCDCA at all measured parameters except the secretion of newly synthesized protein. PMID- 11484844 TI - Hemolytic anemia, thrombosis, and infarction in male and female F344 rats following gavage exposure to 2-butoxyethanol. AB - 2-butoxyethanol (BE; ethylene glycol monobutyl ether) is used extensively in the manufacture of a wide range of domestic and industrial products which may result in human exposure and toxicity. BE causes severe hemolytic anemia in male and female rats and mice. In a recent report, female F344 rats exposed to 500 ppm BE by inhalation and sacrificed moribund on day 4 of treatment exhibited disseminated thrombosis associated with infarction in several organs. In contrast, no such lesions were observed in male rats similarly exposed to BE. Additional studies were therefore undertaken to compare the effects of BE in rats of both sexes. Rats received 250 mg BE/kg/day by gavage for 1, 2 or 3 days and were sacrificed 24 or 48 hr after the last dose. Control rats received 5 ml/kg water. Progressive time-dependent hemolytic anemia--macrocytic, hypochromic, and regenerative--was observed in both sexes of rats exposed to BE. Additionally, BE caused significant morphological changes in erythrocytes, first observed 24 hr after a single dose, including stomatocytosis, macrocytosis with moderate rouleaux formation, and spherocytosis. These morphological changes became progressively more severe as BE dosing continued and included the occasional occurrence of schistocytes and ghost cells, rouleaux formation in rats of both sexes, and an increased number of red blood cells with micronuclei in female rats. Overall, the progression of hemolytic anemia and morphological changes as a function of the number of days of exposure varied with gender and suggested a faster onset of hemolysis in female rats. The range of BE-related histopathological changes noted in both sexes was comparable; however, while these lesions were observed in female rats following a single dose, similar effects were first observed in males after 3 consecutive days of exposure to BE. Pathological changes involved disseminated thrombosis in the lungs, nasal submucosa, eyes, liver, heart, bones and teeth, with evidence of infarction in the heart, eyes, teeth and bones. Hemoglobinuric nephrosis and splenic extramedullary hematopoiesis were also noted. An apparent correlation between the severity of hemolytic anemia and subsequent disseminated thrombosis in BE-treated rats is proposed. Thrombosis may be related to intravascular hemolysis, which could be triggered by procoagulant release and/or alterations in erythrocyte morphology, as well as increased rigidity. PMID- 11484846 TI - Cell membrane labeling of Eimeria tenella sporozoites with the fluorescent dye PKH-67 GL for tracking parasite-host interactions. AB - The fluorescent cell linker dye PKH-67 GL was used as a vital stain for sporozoites of Eimeria tenella for tests on viability, invasion of cultured primary chick kidney cells, flow cytometric analysis and fluorescence microscopy. The effect of PKH-67 GL on sporozoites was tested at a range of concentrations of dye and sporozoites. In flow cytometric analysis, 0.5-40x10(-6) M of PKH-67 GL labeled sporozoites to some degree, with the percentage of labeled sporozoites increasing with higher dye concentrations. The optimum concentration was 2x10(-6) M, allowing easy observation by fluorescence microscopy. Morphological changes in the sporozoite at concentrations greater than 5x10(-6) M were accompanied by loss of viability according to a propidium iodide inclusion assay. Sporozoite penetration of primary chick kidney cells was unaffected by the optimal level of 2x10(-6) M, allowing observation of intracellular activities. Overall, the cell linker dye greatly facilitated observation of E. tenella in vitro and in flow cytometric analysis. PMID- 11484845 TI - Modulation induced by estradiol in the acute phase of Trypanosoma cruzi infection in mice. AB - We investigated the effect of 17beta-estradiol on mice resistant to infection by Trypanosoma cruzi. Infected Balb/C, C3H and C57BL/6 female mice had a longer survival time than males, C57BL/6 showing the highest difference (50% cumulative mortality in females versus 100% in males). This lineage was treated with estradiol (from 0.05 microg to 500 microg/mouse) 1 day before infection. Treatment with 50 microg or 500 microg estradiol/ mouse increased mortality and parasitaemia. Low doses had no effect or tended to reduce both parameters. Given that estradiol presented no in vitro effect on trypomastigotes or epimastigotes, the involvement of a direct hormonal effect on the parasite is improbable. Alterations in the humoral T. cruzi-specific response were also discarded, since the kinetics and concentration of anti-T. cruzi IgG were not affected by the treatment. Females infected during an estradiol-descending phase (meta-oestrus) survived longer than those infected during other phases of the oestrous cycle. We confirmed that estradiol interferes with T. cruzi infection. PMID- 11484847 TI - First description of the male and redescription of the female of Philometra salgadoi Vidal-Martinez et al., 1995 (Nematoda: Philometridae) from the ocular cavity of the marine fish Epinephelus morio in Mexico. AB - A redescription of the female and the first description of the male of the philometrid nematode Philomnetra salgadoi Vidal-Martinez, Aguirre-Macedo et Moravec, 1995 are presented on the basis of specimens collected from the type host, the red grouper. Epinephelus morio (Valenciennes; Pisces: Serranidae), in the Gulf of Mexico off the northern coast of Yucatan, Mexico. The morphology of this species is compared with that of related congeneric species. PMID- 11484848 TI - Plasmodium falciparum transmission intensity and infection rates in children in Gabon. AB - Several factors can determine the outcome of a malarial infection. Studies on susceptibility or resistance to malarial infection can be confounded by differences in transmission. In the present study, the relationship between vector abundance and Plasmodium falciparum infection rate of Gabonese children was studied. Indoor human bait catches were conducted in the houses of two groups of children, those who had been found earlier to be either frequently (> 3 infections per year) or rarely (< 0.5 infections per year) infected with P. falciparum. The human biting rate was 12 and 31 bites per person per night during the dry and the rainy season, with 3% and 16% Anopheles, respectively. Anopheles gambiae and A. moucheti were found to be the only vectors involved in the transmission of malaria in this area. No significant difference in the abundance and the rate of P. falciparum infection of the Anopheles mosquitoes was found among children rarely or frequently infected. Differences in transmission cannot account for differences in infection rates in our study group. Hereditary and immunological factors seem to be the primary determinants for the outcome of malarial infection. PMID- 11484849 TI - Exposure of Blastocystis species to a cytotoxic monoclonal antibody. AB - A recently described cytotoxic monoclonal antibody (mAb 1D5) raised against Blastocystis hominis isolate B, was tested for reactivity with 13 different isolates of Blastocystis. The isolates used were previously isolated from humans, rats and reptiles and were maintained as axenised cultures throughout the course of this study. Five B. hominis isolates (B, C, E, G and H) were found to react with mAb 1D5 in immunoblotting studies and the indirect fluorescence antibody test. The pattern of fluorescence observed for all five isolates was diffuse and patchy. Immunoblotting studies revealed that mAb 1D5 reacted with a 29-30-kDa protein found in all five isolates. Results of a cytotoxic assay showed that the mAb exhibited a complement-independent cytotoxic effect on all the exposed isolates. Microscopic observations showed differences in morphology between the Blastocystis cells exposed and unexposed to mAb. Acridine orange staining performed on both exposed and unexposed cells showed similar internal structures when viewed under fluorescence microscopy. PMID- 11484850 TI - Treatment with benznidazole and its immunomodulating effects on Trypanosoma cruzi infected rats. AB - Because benznidazole (BZL) was found to downregulate nitric oxide (NO) and cytokine synthesis by murine macrophages, we analyzed the potential immunological repercussions of BZL treatment in Trypanosoma cruzi-infected rats. To evaluate whether the effects of BZL were also observed in the presence of an immunostimulating cytokine, four groups of acutely infected rats were subjected to one of the following 20-day therapeutic schedules: (1) a curative BZL oral regimen, (2) recombinant interferon (IFN-gamma) injections, (3) a suboptimal BZL regimen (25% of curative dose), (4) the latter plus IFN-gamma. All BZL doses markedly reduced NO-derived metabolites either in the circulation or in cultured macrophage supernatants. This was observed in rats simultaneously treated with IFN-gamma, which contrasted with the augmented NO production seen in animals given this cytokine alone. The untreated rats, and groups receiving monotherapy with IFN-gamma or 25% BZL, had increased circulating interleukin (IL)-1beta and IL-2 levels, which were reduced in those given BZL plus IFN-gamma. Although combined treatment failed to cause the virtually undetectable blood parasite levels induced by optimal BZL doses, chronic myocardial lesions were reduced to the same extent as in those receiving the curative schedule. The beneficial effects of BZL in this trypanosomiasis may also depend on some immunomodulating influences. PMID- 11484851 TI - Eimeria adenoeides sporozoites: enhanced invasion of cultured cells previously inoculated with Eimeria acervulina sporozoites. AB - The effects of prior or concurrent administration of Eimeria acervulina on invasion of cultured cells by Eimeria adenoeides sporozoites and possible mechanisms of action were examined. Baby hamster kidney (BHK) cell cultures that were inoculated with E. acervulina sporozoites were significantly more permissive for invasion by E. adenoeides sporozoites than uninoculated cultures. Enhancement of invasion by E. adenoeides did not occur when the two species were inoculated into cultures concurrently, or within 30 h of each other. However, 48 and 72 h after inoculation of BHK cells with E. acervulina, invasion by E. adenoeides sporozoites was significantly greater than invasion in uninoculated cultures. At 96 h postinoculation with E. acervulina, the enhancing effect on invasion was variable. Culture media collected from E. acervulina-inoculated cultures also significantly enhanced invasion by E. adenoeides. Slight changes in proteins of E. acervulina-inoculated versus uninoculated cell cultures were detected by Western blots of biotinylated and nonbiotinylated cells. Biotinylated bands between 10 and 25 kDa increased in the inoculated cultures. In addition, when chicken anti-E. acervulina sporozoite serum was used as a probe, labeling of a 10 kDa antigen increased in the inoculated cultures. PMID- 11484852 TI - Plasmodium falciparum: in vitro growth inhibition by febrile temperatures. AB - Febrile episodes are the hallmark of malarial infection. We determined the inhibitory effect of febrile temperatures on the in vitro growth of Plasmodium falciparum. Parasites were cultured at various temperatures between 37 degrees C and 40 degrees C for 4 days. A logistic decrease in parasitaemia as a function of temperature was observed for continuous cultures. Incubation of synchronized cultures for different lengths of time during the parasite cycle showed a strong increase of growth inhibition with the maturing of parasites. Febrile temperatures inhibit parasite growth and long, high fevers during malaria may be beneficial for parasite clearance. PMID- 11484853 TI - Increasing the sensitivity of PCR detection in bovine preputial smegma spiked with Tritrichomonas foetus by the addition of agar and resin. AB - Methods for the extraction of DNA from the preputial smegma of cattle infected with Tritrichomonas foetus for the purposes of polymerase chain reaction (PCR) detection are usually time-consuming, relatively insensitive and require hazardous chemicals. In order to solve these problems, we have developed a rapid, sensitive and harmless method to extract quality DNA from preputial smegma spiked with T. foetus. Results indicate that the addition of 5% Chelex-100 resin and 0.05% agar solution to the spiked smegma before the process of DNA extraction by the boiling method can significantly increase the sensitivity of PCR detection. This improved method may be suitable for routine DNA extraction for the diagnosis of cattle and even human trichomoniasis by PCR. PMID- 11484854 TI - Blastocystis hominis: evidence for caspase-3-like activity in cells undergoing programmed cell death. AB - We have shown previously that the human intestinal protozoan, Blastocystis hominis, undergoes apoptosis-like programmed cell death (PCD) when exposed to a cytotoxic monoclonal antibody (mAb), 1D5. In the present study, ELISA and immunoblot assays employing chicken anti-CPP32 antibody suggest that caspase-3 like antigens exist in B. hominis. Using colorimetric and flow cytometric assays for caspase-3 activity, we also observed an increase in activity between 1 h and 6 h after exposure to mAb 1D5, with greatest activity at 6 h. These findings suggest that caspase-3-like proteases play an important role in B. hominis undergoing PCD, similar to the phenomenon in higher eukaryotic organisms. PMID- 11484855 TI - Structure-activity relationships for new organometallic complexes active against bloodstream forms of Trypanosoma brucei brucei. PMID- 11484856 TI - Comparative studies on the levels of serum IgG1 and IgG2a in susceptible B10.BR mice infected with different strains of the intestinal nematode parasite Trichuris muris. AB - Comparative studies were carried out on the levels of serum IgG1 and IgG2a in susceptible B10.BR mice infected with different strains of Trichuris muris (E-J and S strains). As infection proceeded, levels of IgG1 and IgG2a increased in mice infected with either strain until at least day 32 post-infection (p.i.). There were no differences in the IgG1 levels on days 14, 20, and 25 p.i. between mice infected with either E-J or S strain, whereas IgG2a levels on days 20, 25, and 32 p.i. were higher in S-infected mice than those in E-J-infected mice. Isotype switching to IgG2a is entirely dependent on interferon gamma (IFN-gamma) and, according to our previous results, the period of high IFN-gamma production in S-infected B10.BR mice is long compared to E-J-infected B10.BR mice. Thus, increased levels of IFN-gamma may sustain high levels of serum IgG2a in S infected mice. Taken together, levels of serum IgG2a are useful markers of IFN gamma production in T. muris infection. PMID- 11484857 TI - Further remarks on Hammondia hammondi and the taxonomic importance of obligate heteroxeny. AB - In this opinion-paper reasons are given why Hammondia hammondi cannot be considered as a separate species, but should be kept as a species of the genus Toxoplasma, if not a strain of Toxoplasma gondii. PMID- 11484858 TI - Global analysis of the Michaelis-Menten-type ratio-dependent predator-prey system. AB - The recent broad interest on ratio-dependent based predator functional response calls for detailed qualitative study on ratio-dependent predator-prey differential systems. A first such attempt is documented in the recent work of Kuang and Beretta(1998), where Michaelis-Menten-type ratio-dependent model is studied systematically. Their paper, while contains many new and significant results, is far from complete in answering the many subtle mathematical questions on the global qualitative behavior of solutions of the model. Indeed, many of such important open questions are mentioned in the discussion section of their paper. Through a simple change of variable, we transform the Michaelis-Menten type ratio-dependent model to a better studied Gause-type predator-prey system. As a result, we can obtain a complete classification of the asymptotic behavior of the solutions of the Michaelis-Menten-type ratio-dependent model. In some cases we can determine how the outcomes depend on the initial conditions. In particular, open questions on the global stability of all equilibria in various cases and the uniqueness of limit cycles are resolved. Biological implications of our results are also presented. PMID- 11484859 TI - Persistent solutions for age-dependent pair-formation models. AB - Conditions on the vital rates and the mating function are derived which imply existence or nonexistence of exponentially growing persistent age-distributions for age-dependent pair-formation models. PMID- 11484860 TI - Stochastic models of a parasitic infection, exhibiting three basic reproduction ratios. AB - Two closely related stochastic models of parasitic infection are investigated: a non-linear model, where density dependent constraints are included, and a linear model appropriate to the initial behaviour of an epidemic. Host-mortality is included in both models. These models are appropriate to transmission between homogeneously mixing hosts, where the amount of infection which is transferred from one host to another at a single contact depends on the number of parasites in the infecting host. In both models, the basic reproduction ratio R0 can be defined to be the lifetime expected number of offspring of an adult parasite under ideal conditions, but it does not necessarily contain the information needed to separate growth from extinction of infection. In fact we find three regions for a certain parameter where different combinations of parameters determine the behavior of the models. The proofs involve martingale and coupling methods. PMID- 11484861 TI - Approximating the long-term behaviour of a model for parasitic infection. AB - In a companion paper two stochastic models, useful for the initial behaviour of a parasitic infection, were introduced. Now we analyse the long term behaviour. First a law of large numbers is proved which allows us to analyse the deterministic analogues of the stochastic models. The behaviour of the deterministic models is analogous to the stochastic models in that again three basic reproduction ratios are necessary to fully describe the information needed to separate growth from extinction. The existence of stationary solutions is shown in the deterministic models, which can be used as a justification for simulation of quasi-equilibria in the stochastic models. Host-mortality is included in all models. The proofs involve martingale and coupling methods. PMID- 11484862 TI - Predictive factors for disability pension--an 11-year follow up of young persons on sick leave due to neck, shoulder, or back diagnoses. AB - AIMS: Although back diagnoses are recurrent and the main diagnoses behind sickness absence and disability pension surprisingly few longitudinal studies have been performed. This study identifies predictive factors for disability pension among young persons initially sick-listed with back diagnoses. METHODS: An 11-year prospective cohort study was conducted, including all individuals in a Swedish city who, in 1985, were aged 25-34 and sick-listed > or =28 days owing to neck, shoulder, or back diagnoses (n = 213). The following data was obtained: disability pension, emigration, and death for 1985-96, sickness absence for 1982 84, and demographics in 1985 regarding sex, income, occupation, marital status, diagnosis, socioeconomic group, and citizenship. Cox regression and life tables were used in the analyses. RESULTS: In 1996, i.e. within 11 years, 22% of the individuals (27% of the women and 14% of the men) had been granted disability pension. The relative risk for disability pension was higher for women (2.4; p = 0.010), persons with foreign citizenship (3.6; p=0.009), and those who had had >14 sick-leave days per spell during the three years before inclusion, compared to those with <7 days/spell (3.1; p=0.003). CONCLUSIONS: This cohort of young persons proved to be a high-risk group for disability pension. Some of the factors known to predict long-time sickness absence also predict disability pension in a cohort of already sick-listed persons. PMID- 11484863 TI - Caring for relatives with dementia--caregiver experiences of relatives of patients on the waiting list for admission to a psychogeriatric nursing home in The Netherlands. AB - AIMS: Institutionalising a relative is a difficult decision and often relatives have to feel heavily burdened before they take such a step. Then the following delay because of waiting lists can be too much. This paper examines the experiences of caregivers of demented patients at the moment of registration on the waiting list for nursing-home care. METHODS: Interviews were carried out with 93 informal caregivers and analysis was made of the files of patients who were registered on the waiting list for nursing-home admission in Amsterdam in 1997 and 1998. Data collection included the Interview for Deterioration in Daily living activities in Dementia, The Revised Memory and Behaviour Problem Checklist, the Social Support List-Interaction, and three subscales of the Caregiver Reaction Assessment (CRA) scale. RESULTS: At the moment of admission to the waiting list, half of the respondents were rather heavily burdened or worse. Less severe dementia, lower age of the patient and providing more hours of informal care especially accounted for more negative experiences of caregivers. The self-esteem derived from caregiving was higher for caregivers with lower income and for those who perceived the quality of the relationship with the demented person as better. CONCLUSIONS: The high burden levels at the moment the decision to institutionalise the patient is taken put a heavy claim on the energy needed to continue to care during the ensuing waiting period. More social support and formal home care may reduce the level of burden of caregiving. PMID- 11484864 TI - Shifts in B12 opinions in primary health care of Sweden. AB - AIMS: The diagnosis and management of vitamin B12 deficiency varies between countries and within countries. The aim of the study was to map current attitudes and values behind clinical decision-making in Swedish primary health care, which has a unique B12 tradition: two patients out of three are treated with oral high dose cyanocobalamin. Most patients with B12-associated problems are managed in primary health care by general practitioners (GPs). METHODS: The study was designed to elucidate possible opinion shifts among GPs during the period 1996 1998. GPs (n=499), stratified and randomized, received a questionnaire with 24 statements on B12-associated clinical and laboratory problems, to be evaluated by a visuo-analogue scale. RESULTS: The majority of GPs in primary health care in Sweden accepted homocysteine and methylmalonic acid (MMA) as markers for functional deficiency of vitamin B12. The evaluation of classical markers of B12 deficiency was wary and balanced. There was a consensus of the need for B12 therapy to risk groups such as patients with atrophic gastritis or previous gastric surgery. The answers also appeared to reflect an improvement of professional knowledge and competence concerning B12-associated problems among Swedish GPs between 1996 and 1998. CONCLUSIONS: The overriding conclusion was that B12-associated opinions of Swedish GPs were stable within the period studied, with marginal improvements of knowledge and competence. PMID- 11484865 TI - Overutilization of antenatal care in Norway. AB - BACKGROUND: It has long been a common belief in Norway that all pregnant women attend antenatal care, but no documentation has been provided. In 1984, official guidelines were issued recommending a reduction of the number of routine visits. However, no studies have been performed in order to monitor whether the recommendations are followed. AIMS: Utilization review of antenatal care in Norway. METHOD: A national cross-sectional study, comprising all deliveries in all obstetrical units in the country during a two-week registration period in June 1996. Information on onset of antenatal care, the number of visits, parity and gestational age at the time of delivery was collected. The study comprised 1,557 deliveries; 45 of the 60 obstetrical units in the country participated. RESULTS: The mean number of antenatal visits was 12.2. Only two of the 1,557 women (0.1%) delivered without any previous antenatal care. A total of 80% started antenatal care in the first trimester, 0.4% had their first antenatal visit in the third trimester. The mean number of antenatal visits was substantially higher than the recommended number. CONCLUSION: Antenatal care providers do not comply with the official guidelines. PMID- 11484866 TI - The use of analgesics and hypnotics in relation to self-rated health and disability pension--a prospective study of middle-aged men. AB - AIMS: This cohort study on urban middle-aged men investigates the association between the use of analgesics and hypnotics, self-rated health (SRH) and disability pension. METHODS: Five birth-year cohorts of middle-aged, urban, Swedish men were invited to a screening programme and were followed for approximately 11 years. RESULTS: Out of all the subjects (n = 5798), 12.4% received a disability pension during follow-up, 27.0% rated their health as less than perfect, 10.6% used analgesics and 2.9% used hypnotics. Compared with non users of analgesics and hypnotics, the adjusted hazard ratio of disability pension for the simultaneous use of both drugs was 7.0 (95% CI: 4.3, 11.6) and the adjusted odds ratio of poor SRH was 16.5 (6.3, 43.5). Thus, the use of analgesics and hypnotics was positively related to poor SRH and predicted award of a disability pension within an 11-year follow-up. This may reflect that the use of analgesics and hypnotics is a proxy of disease but an independent negative effect on health cannot be excluded. CONCLUSIONS: Information on the use of these drugs could be used to predict the award of a disability pension, such as in different geographical areas or population groups. PMID- 11484867 TI - Risk factors for coronary heart disease in different socioeconomic groups of Lithuania and Sweden--the LiVicordia Study. AB - AIMS: Lithuanian middle-aged men have a fourfold higher risk for coronary heart disease (CHD) mortality compared with Swedish men. In Sweden, CHD mortality is twice as high in blue- compared with white-collar workers. Whether the same risk factors that characterized Lithuanian men, compared with Swedish men, could be found in low socioeconomic groups within the cities was investigated. METHODS: The LiVicordia study compared both traditional and new possible risk factors for CHD among 150 50-year-old men in Linkoping, Sweden and Vilnius, Lithuania. A comparison was made of the prevalence of these risk factors in high and low socioeconomic groups within the cities and, after controlling for the city, variations across socioeconomic groups in the total sample. RESULTS: Small differences were found in traditional risk factors between cities. However, Vilnius men were shorter, had lower serum levels of antioxidant vitamins, more psychosocial strain, and lower cortisol response to a standardized laboratory stress test. These characteristics were also found among men in low social classes in both cities. In linear regression models, short stature, low serum beta-carotene, low social integration, coping and self-esteem, high vital exhaustion, high baseline and low cortisol response to stress were related to low social class. CONCLUSIONS: The same set of risk factors, mainly relating to oxidative and psychosocial stress, that characterized Vilnius men was also found in men in low social classes within the cities. The results suggest that a common set of risk factors may help to explain health differences both between and within countries. PMID- 11484869 TI - Challenges for public health in the global context--prevention and surveillance. PMID- 11484868 TI - Self-rated health as a predictor of disability pension and death--a prospective study of middle-aged men. AB - OBJECTIVE: Self-rated health (SRH) is increasingly attracting attention as a predictor of morbidity and mortality while its relation to impaired function has been given less momentum. The aim of this study was to assess the relation between SRH and the risk of being awarded a disability pension and premature death. METHODS: Five birth-year cohorts of middle-aged men were invited to a screening programme and were followed for approximately 11 years. Of the 718 (12%) men with a disability pension granted during follow-up, 46% had perceived their health as perfect on inclusion. The corresponding figure for the remaining 5,082 men was 77%. RESULTS: The crude relative risk (RR) of disability pension for men with SRH less than perfect was 3.7 (3.2-4.2). After adjustment for premorbidity/medication, the RR declined to 3.3 (2.8-3.8). The crude RR of death associated with SRH less than perfect was 1.6 (1.3-2.0), unchanged after adjustment. CONCLUSION: The results showed that SRH is a strong and independent predictor of disability and, to a lesser degree, of mortality. PMID- 11484870 TI - Person, place and time--but who, where, and when? PMID- 11484872 TI - Conditions during childhood and adolescence as explanations of social class differences in disability pension among young men. AB - AIMS: This study investigates whether conditions present or established in youth and adolescence among young men contribute to the differences in the risk of an early disability pension (DP) among social classes. METHODS: The study is based on data from a nationwide survey of the 49,285 Swedish males born between 1949 and 1951 who were conscripted into military service between 1969 and 1970. Data on socioeconomic groups were based on information of occupation and educational level reported in the census of 1975 held by Statistics Sweden. Potential psychosocial and behavioural risk factors were linked to records from the Swedish Social Insurance Board up until 1993. The analyses were based upon those 33,609 conscripts with information on all background variables who reported an occupation and who were not granted a DP in 1975. RESULTS: The strongest social class difference in the distribution of risk indicators was found for low ranking on the psychometric tests and for having been in a remedial class. In the univariate analyses, the highest odds ratios were noted for unskilled manual workers. In the multivariate model, with all the background variables included, the increased risk ratios for lower socioeconomic groups decreased considerably for a DP irrespective of diagnosis, and diminished for a DP with an alcohol related diagnosis. CONCLUSION: It is concluded that conditions present or established in youth and adolescence are of major importance to understand the strong social class gradient in disability pensions among young men. It is suggested that the increased risks for skilled and unskilled manual workers compared with non-manual employees might be interpreted according to the concept of unfavourable life careers. PMID- 11484871 TI - Juvenile and early adulthood smoking and adult educational achievements--a 31 year follow-up of the Northern Finland 1966 Birth Cohort. AB - AIMS: To investigate the association between juvenile and early adult cigarette smoking and educational achievements up to the age of 31 years. METHODS: As a part of the follow-up of the North Finland 1966 Birth Cohort (n = 10542) from 1966 to 1997, smoking was assessed at the ages of 14 and 31 by postal questionnaires. The highest level of educational attainment was obtained from the National Education Registry of Statistics Finland up to the age of 31 years. RESULTS: Adult smoking (at age 31) and prolonged smoking (at ages 14 and 31) were both associated with an approximately two- to sixfold, adjusted odds for educational underachievement. Smoking only at age 14 showed none of these associations. CONCLUSION: These results are unlikely to be causal, but may be explained by other characteristics associated with smoking such as personality or lifestyle factors, as well as the effect of knowledge related to smoking gained during higher education. PMID- 11484873 TI - Positive correlation between circulating leptin levels and lipid lipoproteins in postmenopausal women administered hormone replacement therapy. AB - Beneficial effects of hormone replacement therapy are reported on plasma concentrations of lipids and lipoproteins. Plasma leptin levels are reported to reflect lipid metabolism. We treated 40 healthy postmenopausal women with continuous combined HRT (0.625 mg conjugated equine estrogen and 2.5 mg medroxyprogesterone acetate orally) daily for 6 months and then investigated total cholesterol, triglyceride, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol, which are considered to be factors inducing cardiovascular disease (CVD). We measured the plasma levels of lipids and leptin, which are considered to be altered with HRT. Plasma leptin and lipid levels were measured at baseline and after 3 and 6 months of HRT. The plasma levels of leptin in PMW were not significantly reduced by HRT. Although the plasma levels of total cholesterol and LDL-cholesterol did not change by HRT, the administration of HRT significantly increased plasma levels of triglycerides and HDL-cholesterol in PMW. The correlation between leptin and total cholesterol or triglycerides was positive after HRT while these relations were not correlated at baseline. The correlation between lipid levels and leptin may explain the new role of leptin in plasma lipid levels in HRT. PMID- 11484874 TI - Gene expression of heme oxygenase-1 during glial activation by lipopolysaccharide. AB - The effect of bacterial lipopolysaccharide (LPS) on the expression of mRNA encoding heme oxygenase-1 (HO-1), which is the rate limiting enzyme in heme catabolism and is also known as heat shock protein 32 (HSP32), was examined in primarily cultured rat glial cells. Treatment of cells with LPS elicited an increase in HO-1 mRNA, accompanying down regulation of delta-aminolevulinate synthase, in a dose-dependent fashion. HO-1 mRNA increased markedly at 12 h after LPS treatment (10 microg/ml) and reached a maximum at 24 h. In contrast, HSP70, a major heat shock protein, slightly increased only at 6 h after LPS treatment and returned to the control level by 12 h. These results suggest that HSPs are induced under separate regulation during glial activation by LPS through oxidative stress, a part of which is likely mediated by intracellular free heme. PMID- 11484875 TI - In vivo imaging of increased oxidative stress in the liver by electron spin resonance-computed tomography. AB - The aim of this study was to investigate whether increased hepatic oxidative stress could be visualised in living animals before the onset of obvious liver injury. Acute hepatic injury was induced in mice by priming with heat-killed Corynebacterium parvum followed by injection of a low dose of lipopolysaccharide (LPS). Low frequency band electron spin resonance-computed tomography (ESR-CT) with 3-carbamoyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (carbamoyl-PROXYL) was used to visualize hepatic oxidative stress. Biochemical and histological investigations performed 3 h after injection of LPS revealed no obvious injury to the liver. Conversely, significant hepatic oxidative stress could be detected at this time. Nitroxides such as carbamoyl-PROXYL are rapidly reduced to the corresponding hydroxylamine in vivo. resulting in the disappearance of their ESR signals. The kinetic clearance of carbamoyl-PROXYL after intravenous administration was delayed significantly in mice that had received LPS, due to impairment of the reduction system by hepatic oxidative stress. ESR-CT of the murine abdomen revealed a high intensity area of carbamoyl-PROXYL which consisted mainly of the liver and enlarged spleen. Time-course observations with ESR-CT using carbamoyl-PROXYL showed that the high intensity area in the liver disappeared rapidly due to reduction of carbamoyl-PROXYL. Three hours after LPS injection into the same mouse, ESR-CT images were obtained again by intravenous injection of carbamoyl-PROXYL. The ESR-CT images of the mouse with hepatic oxidative stress clearly showed that the high intensity area of carbamoyl-PROXYL in the liver persisted for a long period of time. This study is the first report to describe the use of in vivo ESR-CT for visualizing the state of increased oxidative stress in the liver before the onset of obvious hepatic injury. PMID- 11484876 TI - Distinct mechanisms of transport of ascorbic acid and dehydroascorbic acid in intestinal epithelial cells (IEC-6). AB - Ascorbic acid (AsA) is an essential nutrient for humans as they lack its biosynthesizing key enzyme. Its absorption mechanism in small intestinal epithelial cells still remains to be resolved. In this study, the transport mechanisms functioning on the uptake of AsA and its oxidized form, dehydroascorbic acid (DHA), were investigated using rat small intestinal epithelial cell line IEC-6. Both AsA and DHA were accumulated in the cells in time- and concentration-dependent manners, but their absorption kinetics were apparently different. The saturability of AsA uptake was shown at a considerably lower concentration in IEC-6 cells as well as other mammalian cells, indicating that this absorption was mediated by a specific transporting carrier. The absorption efficiency of AsA was about 1/5-1/10 that of DHA at the same concentration range and, moreover, the uptake of DHA was almost comparable to that of 2-deoxy-D-glucose, an alternative of glucose. The uptake of AsA was diminished by the removal of sodium ion, but not by the addition of glucose, whereas that of DHA was sodium ion-independent and effectively inhibited by glucose. In addition, phlorizin and cytochalasin B, which are blockers of glucose transporters, interfered the uptake of DHA more efficiently than that of AsA. These results indicate that there are at least two distinct transport systems of vitamin C in rat small intestinal epithelial cells; AsA is transported by a specific transporter and DHA is mainly transported by glucose transporter(s). PMID- 11484877 TI - Circadian changes in the pharmacokinetics of triamcinolone acetonide in rabbits. AB - Circadian changes in the pharmacokinetics of triamcinolone acetonide were investigated in rabbits after a single intravenous administration of the drug, 1.0 mg/kg, at 08:00 and 20:00. At 20:00 hrs, the area under the plasma concentration-time curve from time zero to time infinity was significantly greater (46% increase), and this could be due to significantly slower total body clearance (31% decrease) than that at 08:00. The terminal half-life was significantly longer (82%) at 20:00. However, the apparent volume of distribution at steady state and total amount of unchanged drug excreted in 12 hr urine were not significantly different between two groups of rabbits. PMID- 11484878 TI - Induction of a novel gelatinolytic activity in synovial tissue of patients with rheumatoid arthritis. AB - Gelatinolytic activity induced by longtime incubation at 37 degrees C was found in extracts of rheumatoid synovial tissues. The enzyme activity was assessed by gelatin zymography and 3H-gelatin degradation assay. The observed enzyme had different characteristics from matrix metalloproteinases; it did not require metal ions for activity. However, metallocheltors blocked activation and addition of some metal ions restored the activation. The molecular size of the enzyme was changed time-dependently. The approximate molecular weight of the first enzyme produced by incubation was 65 kDa and it was converted to a broad size molecule with a molecular weight of 50 kDa after further incubation. Substrate specificity was detected for denatured collagen types I, II, III and IV. PMID- 11484879 TI - Scanning electron microscopic findings of cleaved unfertilized eggs ovulated by repeated administration of gonadotrophin. AB - Oocytes in the degenerative process are suggested to be rescued and to developed to ovulation by repeated induction of superovulation using gonadotrophin (GTH). In this study, the morphology of abnormally cleaved unfertilized eggs obtained by repeated induction of superovulation in mice using PMSG/hCG was observed by scanning electron microscope. As a result, loss of microvilli was noted on the cell surface of cleaved unfertilized eggs. On the other hand, ample microvilli were observed on the cell surface of fertilized eggs in the morula stage obtained 65 hours after spontaneous copulation used as controls. Loss of microvilli on the cell surface is a characteristic finding in apoptotic cells. Cleaved unfertilized eggs obtained by repeated induction of superovulation using GTH are considered to be in the degenerative process due to apoptosis. PMID- 11484880 TI - Na+/H+ exchange inhibitor reduces vascular injury caused by ischemia-reperfusion in the rat hind limb. AB - This study was designed to evaluate the effects of an Na+/H+ exchange inhibitor, SM-20550, on ischemia-reperfusion injury in the skeletal muscle. Male Sprague Dawley rats were exposed to ischemia and reperfusion by clamping and releasing clamps both at the abdominal aorta and the bilateral femoral arteries. Rats were divided into three groups; the sham, the SM-20550 treated (SM), and the untreated control (Control) groups. In the SM and control groups, rats were exposed to 5-hr ischemia and 5-hr reperfusion. In the sham group, vessel isolation only was performed. SM-20550 (2.8 mg/kg/hr) in the SM group or vehicle in the sham and control group was continuously administered during ischemia and reperfusion periods. The wall thickness of the vessels were significantly (p<0.01) decreased in the control group than any other group. The internal diameter was significantly (p<0.01) higher in the control and SM group than in the sham group. The wall thickness/internal diameter ratio was significantly (p<0.05) lower in the control group than in the SM group. Thus, an Na+/H+ exchange inhibitor, SM 20550, ameliorated the morphological change due to ischemia reperfusion. These findings provide a clue into the mechanism of ischemia reperfusion injury. PMID- 11484881 TI - Influence of diet and occlusal support on learning memory in rats behavioral and biochemical studies. AB - In order to verify the relationship between tooth loss and the learning memory in rat, male Wistar rats (25 weeks old) were divided into three separate groups: a control group (fed with a solid diet); a soft diet group (fed with a powder diet containing the same components as the solid one) and a molarless group (all molars were removed at 25 weeks and then fed with a powder diet). To evaluate both learning ability and memory, rats were tested with a one-way step through type of passive avoidance apparatus divided into light and dark chambers at 40 weeks. After the passive avoidance test, determination of acetylcholine (ACh) concentration of the cerebral cortex and hippocampus was performed. There was no significant difference between the molarless group and the control group in the response latency before the acquisition trails (non-stimulated period). At day 4 and 7 after the acquisition trials, the response latency of the molarless group was significantly shorter than that in the control group (p<0.05). The ACh levels of the molarless group in the cerebral cortex and hippocampus were significantly lower than that of the control group (p<0.05). It was apparent that tooth loss had an association with a loss of discriminative learning ability. This study suggested that the decrease of masticatory function caused by tooth loss leads to a decrease of ACh synthesis resulting in a learning memory disorder. PMID- 11484882 TI - No effect of water deprivation for 48 hours on the pharmacokiinetics of intravenous tacrolimus in rats. AB - Because the physiological changes that occur in patients with water deprivation could alter the pharmacokinetics of drugs, the pharmacokinetics of tacrolimus were investigated after 1-min intravenous administration of the drug (1 mg/kg) to control rats and rats with water deprivation for 48 h. In rats with dehydration, kidney function seemed to be impaired slightly. Kidney weight (0.800 versus 0.676% body weight) increased significantly and the renal tissue showed only total and mild tubular dilatation and flattening of tubular epithelial cells based on kidney microscopy. However, hepatic function seemed not to be impaired in rats with dehydration. After intravenous administration of tacrolimus, the pharmacokinetic parameters were not significantly different between two groups of rats and the results were expected since tacrolimus was almost completely metabolized in rats (impaired kidney function could not affect considerably the pharmacokinetics of tacrolimus) and hepatic function was not impaired in rats with dehydration. PMID- 11484883 TI - Serum extracellular superoxide dismutase in pediatric patients with various diseases as judged by an ELISA. AB - Extracellular superoxide dismutase (EC-SOD) concentration was measured in sera from 141 patients with 20 forms of infantile diseases including IDDM, SLE and epilepsy, 31 healthy children (controls), and 21 healthy young men by an enzyme linked immunosorbent assay using a polyclonal antibody against human lung EC-SOD. Serum from patients with IDDM and fever of unknown origin had a significantly (p<0.05) lower concentration of EC-SOD than control serum. Part of sera from patients with the seven forms of diseases (SLE, viral infections, epilepsy, nephrosis, hyperthyroidism, hepatic disease, and Reye syndrome), on the other hand, had a greatly high concentration of EC-SOD, albeit not statistically significant. This SOD isoenzyme profile appears to be specific to each pediatric disease. PMID- 11484884 TI - Modeling the transformation of exogenously supplied cAMP in the perfused rat liver. AB - A kinetic model describing the behavior of extracellularly supplied cAMP in the perfused rat liver was derived and compared with experimental data. The model was based on the following conditions and assumptions: a) labeled cAMP is being constantly infused (step input); b) permeation of the cell membrane is an essentially irreversible step (k(in) as transfer coefficient); c) the adenine moiety of cAMP incorporates into a nucleotide pool (km1 as transformation coefficient), which cannot permeate the cell membrane; d) the adenine moiety of cAMP can be transferred from the nucleotide pool to a nucleoside + free base pool (km2 as transformation coefficient), which is able to permeate the cell membrane (k(ef) as transfer coefficient). These events were described by a series of differential equations for which an analytical solution was obtained. Total cellular incorporation of label derived from [3H]cAMP was measured in the isolated perfused rat liver. The equations of the model were fitted to these experimental data by means of a least-squares procedure. In the fitting procedure the previously determined k(in) value (0.55 ml min(-1) ml cellular space(-1)) was used. The model is able to describe the experimental data (correlation coefficient = 0.993 +/- 0.008) with km1, km2 and k(ef) values of 17.11, 0.0948 and 1.385 min(-1), respectively. Simulations revealed the following sequence of decreasing intracellular pool sizes: nucleotide pool > nucleoside + free base pool > intracellular cAMP. The intracellular cAMP concentrations correspond to only 3.2% of the extracellular ones. This low proportion explains why it was generally difficult to detect cAMP in the cell space when this compound was added to an isolated cell system. The model and the parameters determined in the present work can be used to predict intracellular cAMP concentrations in the perfused liver for specific extracellular concentrations. PMID- 11484885 TI - Beta-adrenergic receptors and adenylate cyclase activity in the parotid acinar cells from acute streptozotocin-induced diabetic rats. AB - The changes in beta-adrenergic receptors and in adenylate cyclase (AC) activity were investigated in parotid glands from rats with acute diabetic mellitus (DM) induced by a single injection of streptozotocin (STZ, 80 mg/kg). The animals were divided into three groups: control rats, DM rats, and insulin-treated DM rats. Experiments were performed 7 days after the injection of STZ. Amylase and norepinephrine (NE) contents in parotid glands were markedly decreased in DM rats in comparison with control rats. The density of beta-adrenergic receptor decreased in DM rats, but its affinity for ligand was unaffected. The effect of GTP on isoprenaline (ISO)-stimulated adenylate cyclase (AC) activity significantly decreased in DM rats, but forskolin-stimulated AC activity was unaltered. In addition, diabetes induced the blunted response of AC activity to ISO. The changes in AC activity and in amylase content induced by diabetes were restored by insulin, but those in NE content and receptor density could not. These observations indicate that diabetes decreases NE and amylase contents, receptor density, and receptor-AC coupling in parotid gland, and that these changes would occur in the earlier stage of acute STZ-induced diabetic state. PMID- 11484886 TI - Enhancement of dopamine release by propylene glycol in PC12 cells. AB - Propylene glycol (PG) is widely used as a solvent for injections. However, there are a few reports describing severe toxic effects of PG on human nervous tissues. To elucidate its mechanism, the present study has been conducted to determine whether PG enhances the release of catecholamine in PC12 cells. When the incubation time was longer than 3 min, PG significantly facilitated the dopamine release. PG (0.2-20 %v/v) concentration-dependently increased the dopamine release and the effects of PG at the concentrations above 1% were significant. High K+ (50 mM) and carbamylcholine (50 microM) increased the dopamine release. High K+ and electrical stimulation augmented the action of PG. Tetrodotoxin (1 microM) had no effect on the PG action. In conclusion, PG enhances the dopamine release. It is suggested that the facilitation of the transmitter release from the motor nerve terminals may be related to the PG-evoked skin twitch. PMID- 11484887 TI - Effect of age on rat aortic responses to acetylcholine and nitric oxide donor (NOC-18). AB - The purpose of the present study was to examine the effects of age on vascular responses to acetylcholine (ACh) and NO-releasing compound (NOC-18). The studies were performed in young (4 months old, n = 8) and old (22 months old, n = 6) male rats. Responses to ACh and NOC-18 were examined in vitro by using isolated abdominal aortic rings. The maximum relaxation response to ACh, an endothelium dependent vasodilation, was lower in aortas from old rats. Sensitivity (mean effective concentration; EC50) of ACh in old rats was significantly less than in young rats. There were no differences in maximum NOC-18-induced relaxation, an endothelium-independent vasodilation, in aortas from young and old rats. On the other hand, the concentration-response curve for NOC-18 was shifted to the right and the sensitivity (EC50 to NOC-18) was lower in old rats. These results indicated that both endothelium-dependent vasodilation induced by ACh and endothelium-independent vasodilation induced by NOC-18 are impaired in aorta from old rats. PMID- 11484888 TI - Keep your teeth if you want to talk thyroid. PMID- 11484889 TI - Effects of thyroid hormones on cardiac structure: a tissue characterization study in patients with thyroid disorders before and after treatment. AB - Experimental evidence suggests an involvement of thyroid hormones in myocardial nonmyocyte component growth. We evaluated the possible role of thyroid hormones in myocardial remodeling by ultrasonic tissue characterization (videodensitometry) in 8 hyperthyroid patients, in 10 hypothyroid patients, and in 2 patients with thyroid hormone resistance syndrome (RTH), before, 60, and 120 days after treatment (T0, T60, T120), and in 10 age-matched euthyroids. According to a previously described procedure, the derived collagen volume fraction (dCVF%, an echocardiographic index estimating the collagen content) was predicted from the pixel-level frequency distribution width (broadband, Bb) of the selected echocardiographic images. Thyrotropin (TSH), free thyroxine (FT4), and free triiodothyronine (FT3) were assessed by immunometric method. QT interval dispersion (QTd) on basal electrocardiogram was measured as a marker of dyshomogeneous ventricular repolarization. At T0, Bb and dCVF% were normal in hyperthyroid and euthyroid patients, and slightly increased in RTH patients, whereas significantly higher values were found in hypothyroids. At T60, a significant reduction in Bb was observed in hypothyroids, with nearly normal dCVF% values. This trend was confirmed at T120 with complete normalization of echoreflectivity. No echoreflectivity changes were observed in hyperthyroid and RTH patients during treatment. QTd was significantly increased in hypothyroids at T0, while no significant differences were found among groups at T60 and T120. Because the different videodeonsitometric myocardial properties observed in hypothyroid versus hyperthyroid patients correspond to an increase of dCVF%, this study suggests that thyroid hormones exert an inhibitory effect on myocardial collagen synthesis in humans. PMID- 11484890 TI - Expression of CD30 ligand and CD30 receptor in normal thyroid and benign and malignant thyroid nodules. AB - Because the CD30 ligand (CD30L)/CD30 receptor (CD30) system is expressed in certain malignancies, but has not been studied in thyroid nodules, we investigated its immunohistochemical expression in 6 normal thyroids (NT) and 131 thyroid nodules: 28 colloid nodules (CN), 45 adenomas (15 oncocytic [OA], 30 follicular [FA]) and 58 carcinomas (15 follicular [FTC], 1 insular [ITC], 6 anaplastic [ATC], 30 papillary [PTC], and 6 medullary [MTC]). NT and CN expressed neither CD30L nor CD30 (CD30L-/CD30-). Forty percent of OA and 20% of FA showed epithelial coexpression of CD30L and CD30, and interstitial expression of CD30L, which was also observed in the surrounding normal tissue. Within malignancies, epithelial coexpression of CD30L and CD30 was observed in 7% of FTC, 33% of ATC, 67% of PTC, and 67% of MTC. Only PTC and MTC showed epithelial expression of CD30L in the perinodular tissue with similar frequency (80% PTC, 75% MTC). PTC and MTC had the highest proportion of CD30L+ or CD30+ cells, and together with OA, a thus far unreported nuclear location of CD30L. In PTC, the proportion of CD30L+ cells and the prevalence of nuclear location of CD30L correlated inversely and directly, respectively, with aggressiveness. In conclusion, CD30L/CD30 signaling is activated only past the colloid nodule stage, most frequently in an autocrine fashion. PMID- 11484891 TI - Transient activation of c-Jun NH2-terminal kinase by growth factors influences survival but not apoptosis of human thyrocytes. AB - Activation of c-Jun NH2-terminal kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family, is involved in apoptosis or cell proliferation. We have previously demonstrated that ionizing radiation or thyroid-stimulating hormone activated JNK without linking to thyroid cell apoptosis. To clarify the involvement of JNK activation in thyroid cell survival, we investigated the effects of various growth factors on induction of JNK activation in cultured human thyroid cells. JNK activation was observed at 30 minutes after fetal bovine serum (FBS) stimulation and returned to basal level at 240 minutes. Epidermal growth factor (EGF), transforming growth factor-beta (TGF-beta) and hepatocyte growth factor (HGF) also induced JNK activation, but did not trigger apoptotic cell death. Furthermore, we observed high basal activation of JNK in four of five human thyroid cancer cell lines. Overexpression of c-Met, an HGF receptor, was observed in two of the four cell lines with high basal JNK activity. Our results suggest that JNK activation does not induce apoptosis but is associated with survival or transformation of human thyroid cells. PMID- 11484892 TI - Prognostic factors in well-differentiated follicular-derived carcinoma and medullary thyroid carcinoma. AB - This paper reviews both clinicopathologic and biological factors in differentiated thyroid carcinoma of follicular and C-cell derivation. The value of specific histological parameters including tumor subtypes, vascular invasion as predictors of prognosis in these tumors is discussed and the pertinent literature reviewed. The utility of ploidy analysis and immunohistochemical assessment of hormonal and tumor markers are described. Current information on biological and molecular markers that may be helpful in predicting behavior is discussed. The commonly used staging systems for thyroid cancer are briefly described. PMID- 11484893 TI - Radionuclide-labeled somatostatin analogues for diagnostic and therapeutic purposes in nonmedullary thyroid cancer. AB - Despite the fact that several recent studies report an expression of somatostatin receptors in nonmedullary thyroid cancer (non-MTC), there is still no consensus concerning the diagnostic and therapeutic usefulness of radionuclide-labeled somatostatin analogues in non-MTC. We present the results of 50 scintigraphic studies with (111)In-Pentetreotide ((111)In-P) in 48 patients with metastasizing non-MTC (n = 9 papillary, n = 9 follicular, n = 29 Hurthle cell, n = 1 insular carcinoma). The findings were compared with histology and with other imaging modalities. (111)In-P provided unequivocally positive results in 37 of 50 (74%) of the patients (27% in the 11 patients with current thyroglobulin levels <10 ng/mL and 85% in the patients with thyroglobulin >10 ng/mL). Histopathology demonstrated that maximal uptake was observed in Hurthle cell carcinoma (95% positive examinations if thyroglobulin exceeds 10 ng/mL). We also describe for the first time dosimetric and clinical data from the courses of 90Y-DOTATOC therapy in three patients with progressive, somatostatin-receptor-positive non MTC (up to 9.3 GBq per 4 cycles). Tumor progression could not be stopped in any of the patients treated with 90Y-DOTATOC. We conclude that (111)In-P is a promising tool for whole-body diagnosis in nonradioiodine-accumulating non-MTC, especially in Hurthle cell cancer, and if 2-[18F]fluorodeoxyglucose-positron emission tomography (FDG-PET) is not available. Although the number of patients treated with 90Y-DOTATOC is still limited, our applied treatment protocol appears to be ineffective in metastasizing non-MTC. PMID- 11484894 TI - Searching for iodine deficiency disorders in schoolchildren from Brazil: the Thyromobil project. PMID- 11484895 TI - Transient stimulatory effects on pituitary-thyroid axis in patients treated with interleukin-2. AB - It has been shown that various cytokine therapies may influence thyroid hormone parameters that may lead to serious side effects including nonthyroidal illness. Interleukin-2 is effective in increasing CD4-T cell numbers in human immunodeficiency virus (HIV)-infected patients and it is used in the treatment of various malignant tumours. However, the association of interleukin-2 (IL-2) therapy and thyroid function is not clearly established as serial systematic measurements of thyroid parameters have not been performed with interleukin-2 as the sole therapeutic agent. Therefore, it was the aim of this study to examine prospectively the impact of a 5-day interleukin-2 therapy on thyroid parameters in asymptomatic HIV-infected patients. Twenty male euthyroid patients (mean age, 42.6 +/- 3.2 years; body weight, 73.4 +/- 3.0 kg) received 9,000,000 IU/d interleukin-2. Thyroid function was evaluated by measurements of serum thyrotropin (TSH), triiodothyronine (T3), thyroxine (T4), free thyroxine (FT4), reverse T3 (rT3), thyroglobulin (Tg), thyroxine-binding globulin (TBG), and anti thyroid-peroxidase (TPO)-antibodies from day 1-4 and on days 7, 14, 20, 40, 60, 80, and 100. All results are given as mean +/- SD. On day 4, we observed a significant increase that was still within normal range of T4 and T3 (p < 0.05). TSH increased from 1.33 +/- 0.57 to 4.53 +/- 1.39 mU/l (p = 0.0001) and FT4 from 18.1 +/- 4.2 to 48.9 +/- 10.9 pmol/L (p = 0.0001) on day 4 with a gradual decrease thereafter. Normalization to baseline levels for TSH (1.45 +/- 0.75 mU/L) and FT4 (18.1 +/- 3.0 pmol/L) was achieved only on day 14. The increase of FT4 was more pronounced (well in the hyperthyroid range) than the increase in total T4 in the presence of normal TBG and albumin concentrations whereas TBG was not affected. We did not observe changes in anti-TPO-antibody levels up to day 100. Our data clearly demonstrate that the administration of interleukin-2 has a stimulatory effect on the pituitary-thyroid axis. The increase of TSH suggests a central stimulation directed by the action of IL-2 as the major mechanism. PMID- 11484896 TI - Estimation of total body iodine content in normal young men. AB - Total body iodine content was estimated in six normal young men, who underwent 125I balance studies during 64-92 days of daily 125I administration. Total body retention of 125I was measured as the difference between total administered 125I and that collected in the urine and feces. Extrathyroidal 125I was the difference between total and thyroidal 125I content. The time-activity data for the ratio of extrathyroidal to total retained 125I were fitted to a growth (inverse exponential) function. Fits of this growth function to the individual data sets yielded asymptotes, the equilibrium extrathyroidal/total 125I ratios. The slopes of this function predicted the time that would have been required to achieve 125I/127I equilibration (approximately 10 months). Geometric mean for the asymptotic extrathyroidal/total 125I ratio was 0.34 (range, 0.19-0.63), if it was assumed that measured urine and feces represented all of the 125I lost to the body. If 90% measurement of 125I loss was assumed, the geometric mean ratio was 0.32 (range, 0.17-0.60). Assuming that 90% of total loss is reflected in measured excreta and that total iodine content of the thyroid gland is 10 mg, geometric mean for total body iodine in these subjects was 14.6 mg (range, 12.1-25.3 mg). PMID- 11484897 TI - Anatomical distribution and sclerotic activity of bone metastases from thyroid cancer assessed with F-18 sodium fluoride positron emission tomography. AB - Currently, bone scintigraphy (BS) is considered to lack sensitivity in detecting bone metastases (BM) from thyroid cancer. We evaluated the anatomical distribution and metabolic behavior of BM as well as the accuracy of BS with and without combination of whole-body iodine scintigraphy (WBI) in detecting metastatic bone disease in thyroid carcinoma. F-18 positron emission tomography (PET), x-ray, BS, and WBI were performed in 35 patients with known or suspected bone metastases from papillary (9 patients) or follicular (26 patients) thyroid carcinoma. Twenty-two metastases were previously known in 14 patients. The indication was staging in 21 patients with high risk for BM, elevated thyroglobulin (Tg)-levels or evaluation of exact extent of BM (14 patients). In addition, results of WBI (35 patients), X-ray (35 patients) F-18 PET (35 patients), MRI of the spine (13 patients), and FDG-PET (15 patients) as well as the clinical course (1.5-4 years) were correlated. BM were detected in 18 patients. Solitary, bifocal, or multiple lesions were present in 9, 2, and 7 patients, respectively. The anatomical distribution of BM (n = 43) was as follows: spine, 42%; skull, 2%; thorax, 16%; femur, 9%; pelvis, 26%; humerus and clavicle, 5%. Sensitivity of BS in interpreting patients as positive or negative for having BM was 64%-85% (specificity, 95%-81%). The combination of BS and WBI was 100% sensitive in detecting metastatic bone disease. One patient had a single BM that was positive at BS but negative on WBI. All metastases were osteolytic on x-ray and two-thirds presented a missing or very limited osteosclerotic bone reaction on F-18 PET. Our data confirm the limited sensitivity of planar BS in detecting BM from thyroid cancer. The combination of BS and WBI, however, was highly accurate. Compared to other malignancies, the distribution pattern of BM presented a lower percentage of vertebral metastases and more patients with single metastases. Those findings in combination with a missing or only slight osteosclerotic bone reaction explain the limited sensitivity of planar BS alone. PMID- 11484898 TI - Congenital goiter with hypothyroidism caused by a 5' splice site mutation in the thyroglobulin gene. AB - In this work we have extended our initial molecular studies of a consanguineous family with two affected goitrous siblings (H.S.N. and Ac.S.N.) with defective thyroglobulin (Tg) synthesis and secretion because of a homozygotic deletion of a fragment of 138 nucleotides (nt) in the central region of the Tg mRNA, identified previously in H.S.N. In order to identify the intron/exon boundaries and to analyze the regions responsible for pre-mRNA processing corresponding to a 138 nt deletion, we performed a screening of a human genomic library. The intron/exon junction sequences were determined from one positive clone by sequencing both strands of the DNA template. The results showed that the deletion mapped between positions 5549 and 5686 of the Tg mRNA and corresponded to exon 30. The positions of the exon limits differed by three nucleotides from the previously reported data obtained from direct sequencing of the deleted reverse transcriptase polymerase chain reaction fragment from H.S.N. These variations are because the intron/exon junctions in this region were not available at the time when the deletion was first described. The deletion does not affect the reading frame of the resulting mRNA and is potentially fully translatable into a Tg polypeptide chain that is shortened by 46 residues. The same 138 nt deletion was observed in reverse transcriptase-polymerase chain reaction studies performed in the thyroid tissues from Ac.S.N. Genomic DNA analysis showed that a G to T transversion was observed at position +1 in the donor site of intron 30. Both affected patients (H.S.N. and Ac.S.N.) are homozygous for the mutation whereas the normal sister (At.S.N.) had a normal allele pattern. The functional consequences of the deletion are related to structural changes in the protein molecule that either could modify the normal routing of the translation product through the membrane system of the cell or could impair the coupling reaction. Probably the mutant Tg polypeptide might be functionally active in the production of thyroid hormone, because in the presence of a normal iodine ingestion (approximately 150 microg/day), Ac.S.N. was able to maintain normal serum levels of total triiodothyronine (T3) associated with relatively low serum total thyroxine (T4) with normal somatic development without signs of brain damage. PMID- 11484899 TI - Atypical subacute thyroiditis: preliminary observations. AB - Nine patients with painless or minimally painful subacute thyroiditis were seen between late June and October 2000. Six had a history of antecedant viral symptoms. Thyroid peroxidase antibodies were negative in eight patients tested; none had a family history of autoimmune thyroid disease. It is possible that these patients represent examples of postviral painless subacute thyroiditis (atypical subacute thyroiditis). In order to establish the nature of the syndrome, cytological examination, HLA typing, and long-term follow-up are necessary. PMID- 11484900 TI - Langerhans cell histiocytosis of the thyroid: a report of two cases and review of the literature. AB - Langerhans cell histiocytosis (LCH) is a monoclonal disease of histiocytes that can involve many or very few organ systems. It is a relatively benign disorder with a 3% mortality in adults. LCH rarely involves the thyroid gland. We report two cases, both presenting in males with a goiter. Both patients were treated with subtotal thyroidectomy. The first patient also received radiotherapy to his thyroid bed and scalp. We summarize the prior reported cases of LCH involving the thyroid and review the current treatment modalities used for LCH. PMID- 11484901 TI - Images in thyroidology. Onycholysis associated with hypothyroidism. PMID- 11484902 TI - Cyclin D1 in thyroid carcinomas. PMID- 11484903 TI - Distinction between autoimmune and nonautoimmune hyperthyroidism by determination of thyrotropin-receptor antibodies in patients with the scintigraphic diagnosis of disseminated autonomy. PMID- 11484904 TI - Role of metabolism in arsenic toxicity. AB - In humans, as in most mammalian species, inorganic arsenic is methylated to methylarsonic acid (MMA) and dimethylarsinic acid (DMA) by alternating reduction of pentavalent arsenic to trivalent and addition of a methyl group from S adenosylmethionine. The methylation of inorganic arsenic may be considered a detoxification mechanism, as the end metabolites, MMA and DMA, are less reactive with tissue constituents, less toxic, and more readily excreted in the urine than is inorganic arsenic, especially the trivalent form (AsIII, arsenite). The latter is highly reactive with tissue components, due to its strong affinity for sulfhydryl groups. Thus, following exposure to AsV the first step in the biotransformation, i.e. the reduction to AsIII, may be considered a bioactivation. Also, reactive intermediate metabolites of high toxicity, mainly MMAIII, may be formed and distributed to tissues. Low levels of MMAIII and DMAIII have been detected in urine of individuals chronically exposed to inorganic arsenic via drinking water. However, the contribution of MMAIIIand DMAIII to the toxicity observed after intake of inorganic arsenic by humans remains to be elucidated. The major route of excretion of arsenic is via the kidneys. Evaluation of the methylation of arsenic is mainly based on the relative amounts of the different metabolites in urine. On average human urine contains 10-30% inorganic arsenic, 10-20% MMA and 60-80% DMA. PMID- 11484906 TI - 2-(Allylthio)pyrazine, a cancer chemopreventive agent, inhibits liver fibrosis induced by dimethylnitrosamine in rats: role of inhibition of transforming growth factor-beta1 expression. AB - Exposure to nitrosamines may be the occupational risk factor for liver cirrhosis. 2-(Allylthio)pyrazine, a chemopreventive agent, inhibits CYP2E1 and induces phase II enzymes. We examined the effects of 2-(allylthio)pyrazine on hepatic fibrosis, a prepathologic state of cirrhosis, and on the expression of transforming growth factor-beta1 induced by dimethylnitrosamine. Treatment of rats with dimethylnitrosamine for 4 weeks increased plasma alanine/aspartate amino transferase and y-glutamyl transpeptidase activities, and bilirubin content, whereas the total plasma protein and albumin levels were decreased. 2 (Allylthio)pyrazine inhibited dimethylnitrosamine-induced increases in the enzyme activities and bilirubin, and restored the plasma protein and albumin contents. Masson's trichrome staining showed that dimethylnitrosamine induced liver fibrosis, the extent of which was reduced by 2-(allylthio)pyrazine treatments. Reverse transcription-polymerase chain reaction analysis revealed that 2 (allylthio)pyrazine inhibited production of transforming growth factor-beta1 mRNA by dimethylnitrosamine. These results demonstrated that 2-(allylthio)pyrazine might inhibit dimethylnitrosamine-induced liver fibrosis due to suppression of CYP2E1 expression and transforming growth factor-beta1 production. PMID- 11484905 TI - Effect of tiagabine and topiramate on porphyrin metabolism in an in vivo model of porphyria. AB - Administration of many antiepileptic drugs to patients with porphyria can precipitate an acute porphyric crisis. Information on the porphyrogenic activity of new antiepileptic drugs is still limited. In the presented study, the effects of tiagabine and topiramate on porphyrin metabolism were evaluated in an in vivo model of porphyria. Administration of the protoporphyrinogen oxidase inhibitor oxadiazon (12.5 mg/kg/day) for four days to male Wistar rats caused a partial block of porphyrin biosynthesis, thus mimicking the condition of quiescent variegate porphyria. Administration of phenobarbital (75 mg/kg/day) to oxadiazon pretreated rats increased liver porphyrin content, liver porphobilinogen content (means 480 nmol/g, control less than 20 nmol/g) and urinary excretion of porphobilinogen (means 1,000 micromol/l, control less than 20 micromol/l). Tiagabine (75 mg/kg/day) and topiramate (75 mg/kg/day) increased liver porphobilinogen content (means 33 and 53 nmol/g respectively) and urinary porphobilinogen concentration (240 and 490 micromol/l respectively). Similar results were obtained in oxadiazon-treated BALB/c mice. In untreated rats, tiagabine and topiramate caused a moderate increase of hepatic pentoxyresorufin-O dealkylase activity (approximately 100 and 200 pmol/min./mg respectively, controls 15 pmol/min./mg). These data demonstrate that administration of tiagabine or topiramate to oxadiazon-treated animals can provoke a condition resembling an acute porphyric attack and suggest that administration of these drugs to patients with suspected porphyria should be avoided. However, 5-day administration of both tiagabine and topiramate (75 mg/kg) is considerably less porphyrogenic than phenobarbital administered at the same dose. PMID- 11484907 TI - Pharmacological coal tar induces G:C to T:A transversion mutations in the skin of muta mouse. AB - Coal tar is a by-product of the distillation of coal. It consists of a complex chemical mixture of aromatic and aliphatic hydrocabons, with high concentrations of polycyclic aromatic hydrocarbons such as benzo[a]pyrene. We have previously shown that single painting on skin of mice increases the mutation frequency 16 times in murine epidermis cells (Thein et al. 2000). Here, we have determined the mutations by DNA sequencing. Coal tar was found to primarily induce G:C to T:A transversions and one-base pair deletions of G:C base pairs. More than half of the mutations were at CpG sites. The mutational spectrum is in agreement with that of benzo[a]pyrene and other polycyclic aromatic hydrocarbon mixtures. PMID- 11484908 TI - Peripheral versus central muscarinic effects on blood pressure, cardiac contractility, heart rate, and body temperature in the rat monitored by radiotelemetry. AB - Our laboratory has found that the organophosphate pesticide chlorpyrifos elicits an elevation in blood pressure that persists for approximately 24 hr after exposure. Since organophosphate pesticides inhibit acetylcholinesterase activity and cause cholinergic stimulation in the central nervous system and peripheral tissues, we suspect that the hypertensive response from chlorpyrifos is elicited by activation of pressor areas in the brain stem, specifically muscarinic receptors which are known to mediate hypertensive responses. Oxotremorine, a muscarinic agonist, should elicit a blood pressure response similar to organophosphate pesticides. This study used radiotelemetry to assess the effects of oxotremorine on blood pressure, heart rate, core temperature, QA interval (a measure of cardiac contractility), and motor activity in the male, Long-Evans rat. Subcutaneous co-administration of 0.2 mg/kg oxotremorine with 1.0 mg/kg methyl scopolamine (i.e., to block oxotremorine's peripheral effects) caused a marked elevation in blood pressure that developed concomitantly with a 2 degrees decrease in core temperature, 60 beats/min. increase in heart rate, increase in cardiac contractility but no change in motor activity. Overall, blood pressure increased by 19 mmHg from baseline and the response persisted for approximately 12 hr after injection. Methyl scopolamine alone increased heart rate but had no effect on blood pressure, core temperature, and motor activity. Oxotremorine injected without methyl scopolamine led to a relatively minor increase in blood pressure and hypothermia. Overall, central muscarinic stimulation with oxotremorine and methyl scopolamine leads to a vigorous hypertensive response that is associated with increased cardiac contractility, suggesting an increase in cardiac output. Combined central and peripheral cholinergic stimulation following oxotremorine without methyl scopolamine, as would also occur with exposure to chlorpyrifos and other organophosphate pesticides, did not elicit as much of a hypertensive response. This would suggest pathways other than those controlled directly with muscarinic receptors are operative in the development of chlorpyrifos-induced hypertension. PMID- 11484909 TI - A possible mechanism of naproxen-induced lipid peroxidation in rat liver microsomes. AB - Previous papers from our laboratory report that naproxen and salicylic acid induced lipid peroxidation in rat liver microsomes, however, the mechanism is still unclear. In the present paper, ferrous iron release, nicotinamide-adenine dinucleotide phosphate reduced form (NADPH) oxidation and hydrogen peroxide (H2O2) formation have been measured to find out which mechanisms are involved in naproxen- and salicylic acid-induced lipid peroxidation. While the increase of ferrous iron release was observed with high concentrations of naproxen, salicylic acid did not stimulate ferrous iron release. Neither of these drugs stimulated NADPH oxidation and H2O2 formation. However hexobarbital and perfluorohexane, known as uncouplers of cytochrome P450, stimulated microsomal NADPH oxidation, O2 consumption, H2O2 formation and water (H2O) formation involving four-electron oxidase reaction. These results suggest that ferrous iron release contributes to naproxen-induced microsomal lipid peroxidation and that naproxen and salicylic acid are not uncouplers of cytochrome P450. Apparently H2O2 does not play an important role in naproxen- and salicylic acid-induced microsomal lipid peroxidation. PMID- 11484910 TI - Generation of hydroxyl radicals by the antiviral compound phosphonoformic acid (foscarnet). AB - Phosphonoformic acid (foscarnet) is an antiviral agent that is used to treat severe cytomegalovirus infections in AIDS patients. We demonstrate by using the ferrous iron indicator Ferrozine and ascorbic acid (vitamin C) that foscarnet can chelate ferric iron and form a redox-active iron complex. By using the hydroxyl radical indicator coumarin-3-carboxylic acid we found that the foscarnet-Fe3 complex formed can readily catalyze hydroxyl radical (.OH) generation by the Fenton reaction: (Fe2+ + H2O2-4Fe3+ + .OH + -OH) if hydrogen peroxide and ascorbic acid are present. Hydroxylation of coumarin-3-carboxylic acid could be blocked by addition of known hydroxyl radical scavengers such as mannitol, sucrose, glucose and dimethyl sulfoxide. Moreover, by using a DNA nicking assay, we found that foscarnet catalyzed hydroxyl radicals can induce single strand brakes in DNA. The potency of the hydroxyl radicals formed to induce damage could also be demonstrated in a phosphate-free buffer where the hydroxyl radicals formed attacked and liberated phosphate from the foscarnet molecule. Our results indicate that foscarnet catalyzed hydroxyl radical formation might take place during conditions where a peroxide generating system(s), vitamin C and transitions metals are present. PMID- 11484911 TI - Removal of xenobiotic dichlorostearic acid from phospholipids and neutral lipids in cultured human cell lines by B-oxidation and secretion of dichloromyristic acid. AB - Chlorinated fatty acids represent a recently discovered group of potentially hazardous organochlorine pollutants in the environment. The ability of human cells to incorporate and metabolise this type of fatty acids has never been investigated. The aim of the present study was, therefore, to investigate if two human cell lines, INT 407 and SH-SY5Y, incorporate and metabolise extracellular dichlorostearic acid. Cells were incubated with 9,10-dichlorostearic acid for 24 hr, and the amounts of chlorinated fatty acids in cells and culture medium analysed every two days for up to 6 or 10 days. Lipids were separated by solid phase extraction, transesterified to fatty acid methyl esters, and analysed by gas chromatography in combination with a halogen specific detector (GC/XSD). Dichlorostearic acid, dichloropalmitic acid and dichloromyristic acid were found in phospholipids and in neutral lipids of the INT 407 cells. Both cell lines secreted considerable amounts of dichloromyristic acid into the culture medium. Cellular or secreted metabolites shorter than dichloromyristic acid were not found. Taken together, the results suggest that human cells may (1) incorporate chlorinated fatty acids into membrane lipids and storage lipids, (2) metabolise cellular dichlorostearic acid to dichloropalmitic acid and dichloromyristic acid by B-oxidation; but that further metabolism is hindered, possibly because of the chlorine atoms, and (3) remove formed dichloromyristic acid by secretion. The removal of cellular dichloromyristic acid might represent an important cellular defence mechanism and deserves further investigations. PMID- 11484912 TI - Ototoxicity in rats exposed to ortho-, meta- and para-xylene vapours for 13 weeks. AB - Male Sprague-Dawley rats were exposed to ortho-, meta- or para-xylene by inhalation (450, 900 and 1,800 p.p.m., 6 hr/day, 6 days/week for 13 weeks) and sacrificed for morphological investigations 8 weeks after the end of exposure. Brainstem auditory-evoked responses were used to determine auditory thresholds at different frequencies. Among the three isomers studied, only para-xylene produced moderate to severe ototoxicity in rats exposed at 900 and 1,800 p.p.m. Increased auditory thresholds were observed at 2, 4, 8 and 16 kHz in rats exposed to 1800 p.p.m. para-xylene. The auditory threshold shifts (35 to 38 dB) did not reverse after 8 weeks of recovery Moderate and severe losses of outer hair cells of the organ of Corti occurred in animals exposed to 900 and 1800 p.p.m. para-xylene respectively. Thus, the no observed effect level of para-xylene was 450 p.p.m. based on the loss of outer hair cells observed by light and electron microscopy. PMID- 11484913 TI - Chronic ethanol consumption induces gene expression of pancreatic monitor peptide, but not SPINK1/PSTI-56, in rats. AB - The primary factors that predispose humans to the development of alcoholic pancreatitis are unknown. One of the earliest observations in humans in whom this disease develops is pancreatic hypersecretion caused by unknown mechanisms. Messenger RNA (mRNA) differential display was performed in a rat model to investigate the molecular mechanisms associated with ethanol-induced pancreatic hypersecretion. Male Wistar rats were pair-fed Lieber-DeCarli diets with or without ethanol for 7 days or 4 weeks. Total RNA was extracted from the pancreas and its neurohormonal control sites. Differentially expressed complementary DNA (cDNA) tags were isolated, cloned, and sequenced. One 248-bp cDNA was consistently and strongly induced in the pancreata of rats fed ethanol for 4 weeks. The sequence was highly homologous to both rat pancreatic monitor peptide (MP) and pancreatic secretory trypsin inhibitor (PSTI-56), also known as serine protease inhibitor, Kazal type 1 (SPINK1). Confirmatory reverse-transcription polymerase chain reaction showed that PSTI-56 expression remained unchanged, whereas MP mRNA levels were elevated more than four times in the pancreata of ethanol-fed rats. These results indicate that long-term ethanol ingestion increases MP mRNA levels in the rat pancreas. Because MP stimulates cholecystokinin release and cholecystokinin is an important stimulant of pancreatic secretion, the enhanced MP gene expression may contribute to pancreatic hypersecretion. PMID- 11484914 TI - Cholecystokinin-regulated exocytosis in rat pancreatic acinar cells is inhibited by a C-terminus truncated mutant of SNAP-23. AB - INTRODUCTION: Exocytosis is thought to result from the fusion of vesicle and plasma membranes caused by the formation of a trans-complex between proteins of the vesicle-associated membrane protein (VAMP) family on the vesicle with members of the syntaxin and synaptosomal-associated protein of 25 kd (SNAP-25) families on the plasma membrane. In the pancreatic acinar cell, synaptosomal-associated protein of 23 kd (SNAP-23) is the major SNAP-25 isoform expressed in pancreatic acinar cells, but its role in acinar cell exocytosis has not been determined. AIMS: To examine the role of SNAP-23 in regulated exocytosis in acinar cells, we subcloned into adenoviral vectors SNAP-23, SNAP-25, and dominant negative mutants in which the C-terminal domains corresponding to the botulinum neurotoxin A cleavage sites are deleted. METHODOLOGY AND RESULTS: High-efficiency infection of rat pancreatic acini in culture with these adenoviruses by subcellular fractionation showed that the overexpressed SNAP-23, SNAP-25, and their truncated mutant proteins were uniformly targeted to the zymogen granules and plasma membrane. To maximally stimulate apical exocytosis from these infected acini, we used the cholecystokinin-phenylethyl ester analog (CCK-OPE), which does not show inhibition of secretion from maximal levels at high doses. CCK-OPE-stimulated amylase release from adenovirus-cytomegalovirus (AdCMV)-SNAP-23 or AdCMV-SNAP-25 infected acini to the same extent as from acini infected with the empty vector. In contrast, CCK-OPE-evoked enzyme secretion from AdCMV-SNAP-23deltaC8- and AdCMV SNAP-25(1-197)-infected acini were inhibited by 60% and 40%, respectively. The identical targeting of the mutant SNAP-23 and SNAP-25 proteins to the same membrane compartments as SNAP-23 suggests that the inhibition of secretion was a result of their competition against endogenous SNAP-23. This is supported by the fact that this inhibition by the mutant proteins was partially reversed or rescued when the AdCMV-SNAP-25AC8- or AdCMV-SNAP-25(1-197)-infected acini were co infected with wild-type SNAP-23 or SNAP-25. CONCLUSION: From these results, we conclude that SNAP-23 plays a role in CCK-evoked regulated exocytosis in the acinar cells. PMID- 11484915 TI - Effect of camostat mesilate on the expression of pancreatitis-associated protein (PAP), p8, and cytokines in rat spontaneous chronic pancreatitis. AB - An oral protease inhibitor, camostat mesilate (CM) has been used clinically for chronic pancreatitis (CP) in Japan, but it lacks enough scientific evidence of its effectiveness. The aim of this study was to analyze the effect of CM on the gene expressions of pancreatitis-associated protein (PAP), p8, and cytokines such as interleukin-6 and transforming growth factor-beta1 in spontaneous CP model (WBN/Kob rats). CM (10 mg/100 g body weight), mixed in MB-3 diet, was administered orally and gene expressions were analyzed by reverse transcription polymerase chain reaction. In untreated WBN/Kob rats, the gene expressions of all the four factors peaked at 12 weeks, whereas they were significantly suppressed in the CM-treated rats. CM significantly increased the body weight and pancreatic wet weight, and it significantly inhibited inflammatory changes and fibrosis of the pancreas. These results suggest that CM inhibits pancreatic inflammation and fibrosis through the suppression of gene expressions of PAP, p8, and cytokines in CP. PMID- 11484916 TI - Wheat germ extract decreases glucose uptake and RNA ribose formation but increases fatty acid synthesis in MIA pancreatic adenocarcinoma cells. AB - The fermented wheat germ extract with standardized benzoquinone composition has potent tumor propagation inhibitory properties. The authors show that this extract induces profound metabolic changes in cultured MIA pancreatic adenocarcinoma cells when the [1,2-13C2]glucose isotope is used as the single tracer with biologic gas chromatography-mass spectrometry. MIA cells treated with 0.1, 1, and 10 mg/mL wheat germ extract showed a dose-dependent decrease in cell glucose consumption. uptake of isotope into ribosomal RNA (2.4%, 9.4%, and 28.0%), and release of 13CO2. Conversely, direct glucose oxidation and ribose recycling in the pentose cycle showed a dose-dependent increase of 1.2%, 20.7%, and 93.4%. The newly synthesized fraction of cell palmitate and the 13C enrichment of acetyl units were also significantly increased with all doses of wheat germ extract. The fermented wheat germ extract controls tumor propagation primarily by regulating glucose carbon redistribution between cell proliferation related and cell differentiation-related macromolecules. Wheat germ extract treatment is likely associated with the phosphorylation and transcriptional regulation of metabolic enzymes that are involved in glucose carbon redistribution between cell proliferation-related structural and functional macromolecules (RNA, DNA) and the direct oxidative degradation of glucose, which have devastating consequences for the proliferation and survival of pancreatic adenocarcinoma cells in culture. PMID- 11484917 TI - Beneficial effects of growth hormone on bacterial translocation during the course of acute necrotizing pancreatitis in rats. AB - Because bacterial translocation from the gut is one of the important sources of bacterial infection in acute necrotizing pancreatitis (ANP) and growth hormone (GH) has the ability to promote the intestinal epithelial proliferation, we investigated the effects of GH on bacterial translocation in a rat ANP model. ANP was induced in rats by injection of 5% sodium taurocholate into the biliopancreatic duct. The rats with ANP were treated with either human recombinant GH or placebo. Laparotomized animals without induction of ANP (sham operation [SO]) served as controls. At 24 hours after operation, blood was drawn for bacterial culture and determination of amylase, lipase, and endotoxin. Peritoneal fluid and specimens of mesenteric lymph nodes (MLN), liver, pancreas, and spleen were taken for bacterial culture by standard techniques. Intestinal mucosal permeability was assessed by measuring the movement of 125I-labeled albumin from blood to intestinal lumen. Insulin-like growth factor-1 (IGF-1) mRNA was detected in the liver and ileum by reverse transcriptase-polymerase chain reaction (RT-PCR). Morphologic changes of pancreas and ileum were also analyzed. Administration of GH significantly decreased the serum amylase, lipase activities, plasma endotoxin level, and incidence of bacterial translocation. Moreover, the survival rate of ANP rats was improved. The severity of inflammation in pancreas and ileum was alleviated by GH treatment. Ileal mucosal thickness, villus height, and crypt depth in GH treatment rats were obviously increased compared with those of ANP rats. The intestinal permeability was markedly improved in the GH group versus the ANP group. GH treatment resulted in up-regulation of IGF-1 mRNA expression in ileum, but not in liver. These results suggested that exogenous GH had beneficial effects in maintaining the integrity of intestinal mucosal barrier and reducing the incidence of bacterial translocation in rats with ANP. One of the mechanisms might be the up-regulation of IGF-1 mRNA in intestine by GH treatment. PMID- 11484918 TI - Transdifferentiation of human islet cells in a long-term culture. AB - It has been established that ductal cells or precursor cells within the ductal tree of the pancreas can differentiate into islet cells. Although islet cells can also form exocrine cells, it is unclear whether they arise from precursor (stem) cells or from mature endocrine cells by transdifferentiation. Using a defined culture medium and technique for islet purification, for the first time we were able to maintain human islets in culture for more than a year. Multilabeling immunohistochemical and immunoelectron microscopic examination of the islets at different days of culture using islet cell markers (antibodies to hormones, neuron-specific enolase, chromogranin A) and ductal cell markers (cytokeratins 7 and 19, carbonic anhydrase II, DU-PAN2, CA 19-9, and MUC1) revealed that endocrine cells gradually transdifferentiate to ductal, acinar, and intermediary cells. Although islet hormone secretion ceased after day 28 in culture, endocrine cells were still detectable at day 60. However, later, all endocrine and exocrine cells were replaced by undifferentiated cells that expressed neuron-specific enolase, chromogranin A, laminin, vimentin, cytokeratin 7 and 19, alpha-1 antitrypsin, transforming growth factor-alpha, and epidermal growth factor receptor. Our data thus show that, under proper conditions, human islets can be maintained in vitro over a long period and that, in the culture condition, islet cells seem to transdifferentiate to exocrine cells and undifferentiated cells, which may be considered pancreatic precursor (stem) cells. PMID- 11484919 TI - Immunocytochemical localization of prohormone convertase 1/3 and 2 in pancreatic islet cells and islet cell tumors. AB - Peptide hormones are synthesized as bigger prohormones, which are processed posttranslationally into smaller active hormones. Proinsulin and proglucagon are processed into insulin and glucagon by prohormone convertase (PC) 1/3 and 2. The current study was performed to test a hypothesis that there may be some difference in immunoreactive PC levels between normal islet cells and islet cell tumors, as the latter contain more prohormones than the former. All islet cell tumors, including insulinomas, gastrinomas, glucagonomas, pancreatic polypeptide omas (PP-omas), and nonfunctioning islet cell tumors, contain fewer PCs than normal islet cells. The smaller PC levels in islet cell tumors may be responsible for the higher levels of prohormones in islet cell tumors, and the smaller levels of PCs in islet cell tumors may be another distinguishing characteristic of islet cell tumors. PMID- 11484920 TI - Modulation of gap and adherens junctional proteins in cultured neonatal pancreatic islets. AB - Fetal and neonatal pancreatic islets have lower insulin secretory responses compared with adult islets. In culture conditions and after treatment with mammosomatotropic hormones, neonatal islets undergo maturation of the secretory machinery that might involve regulation of cell-cell contacts within the islet. This study is an investigation of the effect of prolonged culturing and in vitro treatment with prolactin on the expression of the gap junction-associated connexin 43 and the adherens junction-associated beta-catenin in cultured neonatal rat islets. Pancreatic islets from neonatal Wistar rats were cultured for 24 hours or 7 days, and the treated group was exposed to 2 microg/mL prolactin daily for 7 days. Connexin 43 and beta-catenin were barely detected at the cell-cell contacts in 24-hour-cultured islets, as revealed by immunocytochemical analysis. Nevertheless, both junctional proteins were well expressed at the junctional region in islet cells cultured for 7 days and showed even greater staining in islets after long-term prolactin treatment. In accordance with the morphologic data, neonatal islets cultured for 24 hours displayed a relatively low level of connexin 43, as determined by Western blot analysis. Culturing for 7 days or combined prolactin treatment induced a significant increase in connexin 43 expression; this was 40% greater in the prolactin-treated group than in the control group. Furthermore, an enhancement of the expression of beta-catenin and translocation of this protein to the cell-cell contact site was also observed in neonatal islets cultured for 7 days compared with those cultured for 24 hours. In vitro prolactin treatment induced even greater expression of beta-catenin in islet cells. A correlation was observed between the increased expression of these junctional proteins and an increase in insulin secretion in cultured neonatal islets. In conclusion, prolonged culturing and in vitro treatment with prolactin induce the modulation of gap and adherens junctional proteins in pancreatic islets, which may be an important event in the in vitro maturation process of neonatal islet cells. PMID- 11484921 TI - Recovery of pancreatic beta cells in response to long-term normoglycemia after pancreas or islet transplantation in severely streptozotocin diabetic adult rats. AB - In the well-established, high-dose streptozotocin diabetic rat model, it is unknown whether normoglycemia after pancreas or islet transplantation may induce the expression of the glucose recognition structures and stimulate the replication of the few surviving pancreatic beta cells. Therefore, the endocrine pancreatic tissue was examined immunocytochemically in streptozotocin-treated major histocompatibility complex congenic Lewis rats at 10 and 100 days after transplantation of whole pancreata or isolated islets implanted under the kidney capsule. In the diabetic state the pancreatic beta cells displayed a weak immunostaining for insulin and glucokinase together with a lack of GLUT2 glucose transporter immunoreactivity in the plasma membrane. Ten days after transplantation, the surviving beta cells had regained their normal immunostaining for insulin and for the glucose recognition structures glucokinase and the A single high dose of streptozotocin causes severe experimental insulin dependent diabetes mellitus in adult rats due to a selective destruction of the pancreatic beta cells in the islets of Langerhans. At doses between 50 and 60 mg/kg of body weight, only very few beta cells survive in the pancreas (1-3). The diabetic state is irreversible and insulin-dependent, thus representing an experimental animal model for type I diabetes (2). Because of the prevailing hyperglycemia, even the few residual beta cells in the pancreas do not function properly and therefore cannot contribute even to a basal supply of insulin to the organism (4). Pancreatic beta cells can function properly in a diabetic organism apparently only after restitution of normoglycemia (5). GLUT2 glucose transporter. One hundred days after transplantation, both of whole pancreas or isolated islets, the number of surviving beta cells in islets of the pancreata of the recipient animals had increased by two- to threefold. The regenerated beta cells were surrounded by a rim of other endocrine cells. The increase in the number of beta cells was not accompanied by signs of neogenesis from ductal structures in the pancreata. The authors' observations support the concept that strict long-term maintenance of normoglycemia through adequate supply of insulin from endocrine grafts is the ideal prerequisite for beta-cell recovery and restitution of the glucose recognition structures, as well as replication of beta cells in pancreata with end-stage diabetic beta-cell destruction after high-dose streptozotocin treatment. PMID- 11484922 TI - Insulin-leptin-visceral fat relation during weight loss. AB - INTRODUCTION: The relation between insulin-leptin-visceral fat axis during weight loss has not been studied previously. AIMS: To evaluate the insulin, leptin, and abdominal adiposity relation during weight loss in patients with upper body obesity. METHODOLOGY: Twenty volunteers (7 men, 13 women) with mean age 50.6+/ 6.3 (SD) and upper body obesity (weight 105.4+/-12.3 kg, BMI 35.9+/-2.5 kg/m2) were recruited. Participants were enrolled in a one-arm clinical study using a calorie-deficient diet and an escalating dose regimen of sibutramine, starting with 5 mg daily and increasing in 5-mg increments to 20 mg per day. Body weight, insulin, leptin, glucose, lipids, abdominal computed tomography (CT), and total body electrical conductance (TOBEC) were measured serially at weeks 0, 4, 8, 12, and 24. RESULTS: Eighteen patients completed the 6-month study: one man and one woman discontinued because of adverse events. With diet and sibutramine, body weight was significantly and continuously reduced throughout the 6-month study. There was a 16.0% (p = 0.0001) reduction in body weight (p < 0.001) and 22.5% (p = 0.0001) decrease in total body fat mass. Abdominal CT scans showed a 28.3% (p = 0.0001) reduction in total abdominal fat, a 26.0% (p = 0.0001) reduction in subcutaneous fat (p < 0.001), and a 31.0% (p = 0.0003) reduction in visceral fat (p < 0.001). There was a 32.0% (p = 0.0008) reduction in leptin levels and 37.9% (p = 0.0001) reduction in insulin levels between baseline and week 4, but no further significant reduction in leptin and insulin levels was observed for the duration of the study. There was a significant correlation between insulin and leptin concentrations throughout the study (p = 0.0001). Leptin was presented as a function of insulin measured at the same time. Significant associations between visceral abdominal fat, subcutaneous fat, and leptin were also observed. CONCLUSION: In this study, we found that leptin and insulin were related in weight loss. The data suggest that insulin may act as a strong regulator of leptin secretion during weight loss and that circulating leptin levels can be predicted by insulin level. Using sibutramine in conjunction with hypocaloric diet reduced body weight and decreased fat mass significantly. Visceral and subcutaneous abdominal fat depots were shown to decrease. Whether sibutramine exerts any selective reduction of visceral abdominal fat as opposed to total body fat mass will require further clinical investigation. PMID- 11484923 TI - Liver injury in acute pancreatitis and mitigation by continuous arterial infusion of an antibiotic via the superior mesenteric artery. AB - Macrophage-derived inflammatory mediators, such as tumor necrosis factor-alpha (TNF-alpha), have been shown to play a central role in aggravation of acute pancreatitis (AP), but little is known about their roles in liver injury. We investigated the pathogenesis of the liver injury in AP and assessed the efficacy of arterial infusion of an antibiotic via the superior mesenteric artery (SMA). Infusion of saline (group A) or imipenem (group B) was started 6 hours after induction of AP in dogs by intraductal administration of autologous gallbladder bile. The survival rate in group B was significantly improved compared with group A. Serum alanine aminotransferase levels in peripheral blood and endotoxin levels in portal vein blood were both significantly higher at 18 hours in group A than in group B. When hepatocytes and Kupffer cells were isolated at 18 hours and cultured for 24 hours thereafter, there was significant exacerbation of the hepatocyte injury and significantly greater production of TNF-alpha in the coculture of hepatocytes and Kupffer cells in group A, indicating that the Kupffer cells had been activated. By contrast, both of these manifestations were significantly mitigated in group B. These findings suggest that TNF-alpha secreted by endotoxin-activated Kupffer cells contributes to liver injury in AP, and that SMA infusion of an antibiotic mitigates the liver injury by controlling endotoxin translocation. PMID- 11484924 TI - Publication rates following pancreas meetings. AB - Publication rates and determinants of publication were studied based on abstracts presented at pancreatic meetings. All abstracts presented at the 1994 and 1995 annual meetings of the European Pancreatic Club (EPC) and the American Pancreatic Association (APA) were followed up by searching MEDLINE. Publication rates were compared using log-rank tests and multiple logistic regression. The prestige of the publishing journals was compared using Kruskal-Wallis tests on scientific impact factors (SIF). Overall, 340 abstracts were presented at the EPC, and 254 were presented at the APA. Of these, 203 (59.7%, EPC) and 138 (54.3%, APA) were later published in peer-reviewed journals. Publication rates did not differ by study type or country region of origin. In addition, median SIFs were similar by conference (APA vs. EPC) and research type (basic science vs. clinical studies) (overall, 1.7). However, North American and North/West European articles were published in higher impact journals as compared with those from other countries. Publication rates and median journal SIFs in pancreas research are similar to those reported from other medical specialty meetings. There is no difference by conference, type of research, or origin (North American vs. European). PMID- 11484925 TI - Feasibility of an endoscopic secretin test: preliminary results. PMID- 11484926 TI - Hydronephrosis associated with acute pancreatitis. PMID- 11484927 TI - Pathogenesis of sludge-related pancreatitis: sonographic implications. PMID- 11484928 TI - Emergency surgery in pancreatic disease. PMID- 11484929 TI - Regulation of the tip-high [Ca2+] gradient in growing hyphae of the fungus Neurospora crassa. AB - Previous work has shown that hyphal elongation in the fungus Neurospora crassa requires a tip-high cytosolic Ca2+ gradient. The source of the Ca2+ appears to be intracellular stores as there is no net transplasma membrane Ca2+ flux at the elongating hyphal tip and modification of ion fluxes across the plasma membrane using voltage clamp is without effect on tip growth. To decode the internal mechanisms which generate and maintain the tip-high Ca2+ gradient we first identified calcium regulators which affect hyphal growth and morphology, then determined how they modify cytosolic [Ca2+] and the actin cytoskeleton using fluorescent dyes and confocal microscopy. Cyclopiazonic acid (a known inhibitor of the endoplasmic reticulum calcium ATPase) inhibits growth and increases cytoplasmic [Ca2+] in the basal region 10-25 microm behind the hyphal tip. 2-APB (2-aminoethoxydiphenyl borate, an inhibitor of IP3-induced Ca2+ release) inhibits hyphal elongation and dissipates the tip-high Ca2 gradient 0-10 microm from the tip. Microinjections of the IP3 receptor agonists adenophostin A and IP3 (but not control microinjections of the biologically inactive L-IP3) transiently inhibited growth and induced subapical branches. IP3 microinjections, but not L-IP3, lowered tip-localized [Ca2+] and increased basal [Ca2+]. Even though their effect on [Ca2+] gradients was different, both cyclopiazonic acid and 2-APB disrupted similarly the normal actin pattern at the hyphal apex. Conversely, disruption of actin with latrunculin B dissipated tip-localized Ca2+. We conclude that the tip high Ca2+ gradient is generated internally by Ca2+ sequestration into endoplasmic reticulum behind the tip and Ca2+ release via an IP3 receptor from tip-localized vesicles whose location is maintained by the actin cytoskeleton. PMID- 11484930 TI - Dual function of rhoD in vesicular movement and cell motility. AB - The trafficking of intracellular membranes requires the coordination of membrane cytoskeletal interactions. Rab proteins are key players in the regulation of vesicular transport, while Rho family members control actin-dependent cell functions. We have previously identified a rho protein, rhoD, which is localized to the plasma membrane and early endosomes. When overexpressed, rhoD alters the actin cytoskeleton and plays an important role in endosome organization. We found that a rhoD mutant exerts its effect on early endosome dynamics through an inhibition in organelle motility. In these studies, the effect of rhoD on endosome dynamics was evaluated in the presence of a constitutively active, GTPase-deficient mutant of rab5, rab5Q79L. As rab5Q79L itself stimulates endosome motility, rhoD might counteract this stimulation, without itself exerting any effect in the absence of rab5 activation. We have now addressed this issue by investigating the effect of rhoD in the absence of co-expressed rab5. We find that rhoDG26V alone alters vesicular dynamics. Vesicular movement, in particular the endocytic/recycling circuit, is altered during processes such as cell motility. Due to the participation of vesicular motility and cytoskeletal rearrangements in cell movement and the involvement of rhoD in both, we have addressed the role of rhoD in this process and have found that rhoDG26V inhibits endothelial cell motility. PMID- 11484931 TI - Cytoskeletal alterations in Dictyostelium induced by expression of human cdc42. AB - The rho family of small G proteins has been shown to be involved in controlling actin filament dynamics in cells. To evaluate the functional overlap between human and Dictyostelium G proteins, we conditionally expressed constitutively active human cdc42 (V12-cdc42) in Dictyostelium cells. Upon induction, cells adopted a unique morphology: a flattened shape with wrinkles running from the cell edge toward the center. The appearance of these wrinkles is highly dynamic so that the cells cycle between the wrinkled and relatively normal morphologies. Phalloidin staining indicates that the stellate wrinkles contain dense actin structures and also that numerous filopods project vertically from the center of these cells. Consistent with the hypothesis that cdc42 induces actin polymerization in vivo, cells expressing V12-cdc42 show an increase in the amount of F-actin associated with the cytoskeleton. This is accompanied by an increase in the association of the actin-binding proteins 34-kDa bundler, ABP-120 and alpha-actinin with the cytoskeleton. In conclusion, human cdc42 has various effects on the Dictyostelium actin cytoskeleton consistent with a conserved role of small GTPases in control of the cytoskeleton. PMID- 11484932 TI - Two subcellular localizations of eIF3 p170 and its interaction with membrane bound microfilaments: implications for alternative functions of p170. AB - We previously identified a 170-kDa protein (p170) highly expressed in lung cancers as the major subunit of the eukaryotic translation initiation factor 3 (eIF3). p170 was recently cloned and little is known concerning its characteristics and subcellular localization. In this paper, we report our surprising findings that about 20% of p170 is associated with membranes while the remaining portion is located in the cytoplasm presumably in the eIF3 complex. We also find that p170 interacts with both endoplasmic reticulum and plasma membranes. The binding of p170 to membranes is through actin filaments, consistent with the fact that p170 contains a spectrin repeat motif that may be involved in actin binding. Furthermore, the cytoplasmic p170 is phosphorylated at serine and threonine residues and the phosphorylation is stimulated by serum. However, the membrane-actin-bound p170 is not phosphorylated. The results obtained in this study suggest that p170 may have other functions in addition to participating in translation initiation. Phosphorylation may play an important regulatory role in the function of p170 in translation initiation and other alternative functions. PMID- 11484933 TI - Reversible nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase upon serum depletion. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH; E.C. 1.2.1.12) functions as a glycolytic enzyme within the cytoplasm, but beside its metabolic function it is involved in early steps of apoptosis, which trigger the translocation of GAPDH into the nucleus. As apoptosis can be induced by serum withdrawal, which otherwise causes cell cycle arrest, the linkage between serum deprivation, cell cycle and nuclear transport of GAPDH has been investigated. The intracellular distribution of GAPDH was monitored by confocal laser scanning microscopy of either immuno-stained NIH 3T3 fibroblasts or of cells overexpressing GFP-tagged GAPDH. Serum withdrawal led to an accumulation of GAPDH in the nucleus. In contrast to investigations published so far, this nuclear translocation was a reversible process: cytoplasmic location of endogenous GAPDH or of GFP-GAPDH could be recovered upon serum addition to arrested cells and was not inhibited by cycloheximide treatment. In addition, the nuclear import upon serum depletion had no influence neither on the catalytic activity nor on the expression level of GAPDH. The nuclear export of GFP-GAPDH in serum-deprived cells could be stimulated by serum or directly by the growth factors EGF or PDGE The transport process is not regulated via an initiation of cell cycle arrest, as olomoucine, which causes G1-arrest neither stimulated nuclear accumulation nor prevented nuclear export after serum addition to serum-depleted cultures. Moreover, SV40 transformed 3T3 cells transported GAPDH into the nucleus upon serum deprivation, though the expression of the viral large T-antigen enabled growth factor independent cell proliferation in this cell line. The recruitment of GAPDH to the cytoplasm upon serum stimulation of arrested cells was not impaired by the inhibition of the MAPK signalling pathway with PD 098059. However, further analysis of the growth factor signalling pathway with specific inhibitors revealed that nuclear export was prevented by LY 294002, an inhibitor of the PI-3 kinase. PI3K links the growth factor signalling pathway with cell death via the repression of an apoptotic inducer. Thus, the nuclear accumulation of GAPDH upon growth factor depletion is a reversible process not related directly to cell cycle and likely triggered by survival signals. PMID- 11484934 TI - Inhibition of multicellular development switches cell death of Dictyostelium discoideum towards mammalian-like unicellular apoptosis. AB - The multicellular development of the single celled eukaryote Dictyostelium discoideum is induced by starvation and consists of initial aggregation of the isolated amoebae, followed by their differentiation into viable spores and dead stalk cells. These stalk cells retain their structural integrity inside a stalk tube that support the spores in the fruiting body. Terminal differentiation into stalk cells has been shown to share several features with programmed cell death (Cornillon et al. (1994), J. Cell Sci. 107, 2691-2704). Here we report that, in the absence of aggregation and differentiation, D. discoideum can undergo another form of programmed cell death that closely resembles apoptosis of most mammalian cells, involves loss of mitochondrial transmembrane potential, phosphatidylserine surface exposure, and engulfment of dying cells by neighboring D. discoideum cells. This death has been studied by various techniques (light microscopy and scanning or transmission electron microscopy, flow cytometry, DNA electrophoresis), in two different conditions inhibiting D. discoideum multicellular development. The first one, corresponding to an induced unicellular cell death, was obtained by starving the cells in a "conditioned" cell-free buffer, prepared by previous starvation of another D. discoideum cell population in potassium phosphate buffer (pH 6.8). The second one, corresponding to death of D. discoideum after axenic growth in suspension, was obtained by keeping stationary cells in their culture medium. In both cases of these unicellular specific cell deaths, microscopy revealed morphological features known as hallmarks of apoptosis for higher eukaryotic cells and apoptosis was further corroborated by flow cytometry. The occurrence in D. discoideum of programmed cell death with two different phenotypes, depending on its multicellular or unicellular status, is further discussed. PMID- 11484935 TI - Anticancer drugs induce necrosis of human endothelial cells involving both oncosis and apoptosis. AB - The endothelium is the first physiological barrier between blood and tissues and can be injured by physical or chemical stress, particularly by the drugs used in cancer therapy. We found that four anticancer agents: etoposide, doxorubicin, bleomycin and paclitaxel induced apoptosis in human umbilical vein endothelial cells (HUVECs) (as judged by DNA fragmentation) with a time- and concentration dependent decrease in bcl-2 protein but without the involvement of p53. As revealed by immunoblotting, bax protein was expressed in HUVECs treated with 1 mg/ml etoposide whereas bcl-2 protein disappeared. Oncosis occurred parallel to apoptosis with the release of lactate dehydrogenase into the supernatant, and, for doxorubicin and etoposide with the inversion of the distribution of angiotensin I-converting enzyme between supernatant and cells. Among the four tested anticancer drugs, only doxorubicin induced an oxidative stress, with significative malondialdehyde production. Thus, human endothelial cells in confluent cultures seem to be in an equilibrium of resistance to apoptosis related to bcl-2 expression; this equilibrium can be disrupted by a chemical stress, such as the antiproliferative drugs known as pro-apoptotic for tumour cells. For doxorubicin and bleomycin, this cellular toxicity can be related to their unwanted effects in human cancer therapy. Low doses of doxorubicin, paclitaxel or etoposide, however, could induce apoptosis of endothelial cells of new vessels surrounding the tumour, thus leading to specific vessel regression with minimal toxic effects for the endothelium of the other vessels. These findings provide evidence of relationships between endothelial toxicity of anticancer drugs and the key role of bcl-2 for resistance of endothelium cells toward apoptosis; moreover lack of p53 and bax in quiescent cells contributes to resistance of endothelial cells to DNA-damaging agents. PMID- 11484936 TI - Bioartificial polymeric materials based on polysaccharides. AB - Bioartificial polymeric materials, based on blends of polysaccharides with synthetic polymers such as poly(vinyl alcohol) (PVA) and poly(acrylic acid) (PAA), were prepared as films or hydrogels. The physico-chemical, mechanical, and biological properties of these materials were investigated by different techniques such as differential scanning calorimetry, dynamic mechanical thermal analysis, scanning electron microscopy, and in vitro release tests, with the aim of evaluating the miscibility of the polymer blends and to establish their potential applications. The results indicate that while dextran is perfectly miscible with PAA, dextran/PVA, chitosan/PVA, starch/PVA, and gellan/PVA blends behave mainly as two-phase systems, although interactions can occur between the components. Cross-linked starch/PVA films could be employed as dialysis membranes: they showed transport properties comparable to, and in some cases better than, those of currently used commercial membranes. Hydrogels based on dextran/PVA and chitosan/PVA blends could find applications as delivery systems. They appeared able to release physiological amounts of human growth hormone, offering the possibility to modulate the release of the drug by varying the content of the biological component. PMID- 11484937 TI - DNA-responsive uniform latex particles based on p-chloromethylstyrene. AB - In this study, DNA binding properties of poly(ethylenimine) (PEI)-attached uniform poly(p-chloromethystyrene) (PCMS) particles were investigated. Spherical PCMS latex particles with an average size of 1.75 microm were obtained by the dispersion polymerization of p-chloromethylstyrene (CMS). PEI was covalently attached onto the PCMS particles via a direct chemical reaction between amine and chloromethyl groups, with the equilibrium binding capacities up to 41 mg PEI/g PCMS. In aqueous media, PEI attached-uniform PCMS particles showed an irreversible aggregation behaviour in the presence of DNA. To predict unknown DNA concentration, the aggregation response of these particles to the presence of DNA was quantified by spectrophotometry. Plain PCMS and PEI attached-uniform PCMS particles were also utilized as sorbents in DNA adsorption experiments conducted at +4 degrees C in a phosphate buffer medium at pH 7.4. DNA immobilization capacities up to 45 mg DNA/g PCMS could be achieved with the PEI attached particles. PMID- 11484938 TI - Water-soluble L-alanine and related oligopeptide conjugates with poly[(R,S)-3 hydroxybutanoic acid] oligomers, synthesis and structural studies by means of electrospray ionization multistage mass spectrometry. AB - The reactions of (R,S) beta-butyrolactone with L-alanine and related oligopeptides (Ala-Ala-Ala) were investigated. The resulting water-soluble oligomers were composed of poly[(R,S)-3-hydroxybutanoic acid] (a-PHB) covalently conjugated to L-alanine and Ala-Ala-Ala oligopeptide. The other chain end was of the carboxylic acid type. The structure of the obtained oligomers was assessed by electrospray ionization multistage mass spectrometry (ESI-MSn) and the respective structural information was completed by IR, NMR, and GPC analyses. The molecular weight and structure of the products could be controlled through reaction conditions. Using this new synthetic approach. a-PHB oligomers with well-defined end groups, as well as respective block copolymers, can be prepared via regioselective ring-opening oligomerization of (R,S) beta-butyrolactone induced by amino acids under their zwitterionic form. PMID- 11484939 TI - Network formation of low molecular weight hyaluronic acid derivatives. AB - The oscillatory and steady shear rheological properties of the benzyl esters of hyaluronic acid (HA), partially esterified (Hyaff 11p50), at low molecular weight (150 kDa) were evaluated and compared to the properties of HA at the same molecular weight. At concentrations up to 40 mg cm(-3) both Hyaff 11p50 solutions and HA solutions, behaved as viscous fluids. At higher concentrations, HA ester solutions exhibited an elastic response typical of weak gels, whereas HA exhibited a viscous behaviour. A solid-like response was also observed by lowering the temperature. These results indicate that hyaluronic acid ester solutions can form a weak gel network. The rheological properties of HA derivatives changed significantly compared to HA solutions. The improved elasticity and residence times of these solutions expand the possible applications of hyaluronic acid in the biomedical field. PMID- 11484940 TI - Centrifugally-spun polyhydroxybutyrate fibres: effect of process solvent on structure, morphology and cell response. AB - The structure and morphology of a novel form of poly(beta-hydroxybutyrate) produced by gel-spinning is described. The entangled fibrous nature of the material, which resembles 'cotton wool' suggests possible functions in wound scaffolding devices. The surface structure and fibre diameter distribution of the fibres have been investigated using phase contrast and scanning electron microscopy. Fibres were found to possess a variety of surface irregularities, such as pores and indentations. with diameters mainly in the range 1-15 microm. Additionally, individual fibres were occasionally found to be fused or forked together with neighbours. The effects of blending with various polysaccharides and of altering the process solvent on fibre morphology were also investigated. Under hydrolytic degradation conditions (pH 10.6, 70 degrees C) the fibres degraded by gradual fragmentation and erosion to fibre fragments, particulate matter and eventually to monomer. Altering the production process influenced both the fibre diameter distributions and surface morphology of the constituent fibres. Mammalian and human epithelial cells were used to study the cellular interaction with the spun fibres. SEM studies show that there is little or no cell adhesion to the unmodified fibres, but surface treatment by means of acid and alkali washes promoted cell proliferation on the materials, probably as a result of the introduction of hydroxyl and carboxyl at the surface. Fabrication of non-woven mats, which were subsequently acid or alkali treated, provided a conventional way of forming a cell-adhesive matrix which may have potential value as a wound scaffold. Neither cell line exhibited any cytotoxic response to these polymers. PMID- 11484941 TI - Poly(epsilon-caprolactone-co-D,L-lactide) /silk fibroin composite materials: preparation and characterization. AB - Poly(epsilon-caprolactone-co-D,L-lactide) copolymers with 10, 30, and 50% by weight of silk particles (size range: 5-250 microm) derived from Bombyx mori were blended in acetone solution. After evaporation of the solvent, the morphology, thermal behavior, and mechanical properties of the composites were examined. The composites were transparent and the silk fibroin particles were homogeneously distributed within the composite structure. The particles appeared as bright reflected images under the optical microscope, suggesting that they were in a crystalline state. DSC thermograms of the composites revealed that the glass transition of the matrix was at ca. -18 degrees C. Degradation of the silk fibroin occurred beyond 270 degrees C. The decomposition temperatures and degradation rate decreased with increasing silk fibroin content as revealed by TGA analysis. FTIR spectra of the composites showed absorption bands at 1730 and 1088 cm(-1) for the copolymer and at 3273 and 1617 cm(-1) for the silk fibroin. Although the characteristic lines of poly(epsilon-caprolactone-co-D,L-lactide) were independent of filler concentration. the absorption bands of the beta-sheet form of the silk fibroin increased slightly due to the interaction of silk fibroin with the copolymer. PMID- 11484942 TI - Long term culture of epithelia in a continuous fluid gradient for biomaterial testing and tissue engineering. AB - Epithelia perform barrier functions being exposed to different fluids on the luminal and basal side. For long-term testing of new biomaterials as artificial basement membrane substitutes, it is important to simulate this fluid gradient. Individually-selected biomaterials can be placed in tissue carriers and in gradient containers, where different media are superfused. Epithelia growing on the tissue carriers form a physiological barrier during the whole culture period. Frequently however, pressure differences between the luminal and basal compartments occur. This is caused by a unilateral accumulation of gas bubbles in the container compartments resulting in tissue damage. Consequently, the occurence of gas bubbles has to be minimized. Air bubbles in the perfusion culture medium preferentially accumulate at sites where different materials come into contact. The first development is new screw caps for media bottles, specifically designed to allow fluid contact with only the tube and not the cap material. The second development is the separation of remaining gas bubbles from the liquid phase in the medium using newly-developed gas expander modules. By the application of these new tools, the yield of embryonic renal collecting duct epithelia with intact barrier function on a fragile natural support material can be significantly increased compared to earlier experiments. PMID- 11484943 TI - 'Electrosmog': what is really behind the fight between Italian Ministers and the Vatican? Smoke again? PMID- 11484944 TI - Europe and euthanasia. PMID- 11484945 TI - Not everyone is perfct: about off-patent drugs. PMID- 11484946 TI - Tobacco smoking and prostate cancer: time for an appraisal. PMID- 11484947 TI - Emerging therapies in non-small-cell lung cancer. AB - Lung cancer remains the single most devastating cause of cancer-related death with approximately 1.5 million cases of lung cancer expected worldwide and more than 1.3 million cancer-related deaths in 2001. In the United States alone, of 164,100 news cases expected in the year 2000, about 70,000 will be metastatic disease (stage IV), and another 70,000 will be locally advanced (stages IIIA and IIIB). Therefore, the five-year survival rate for lung cancer has improved only incrementally from 5% in the late 1950s to 14% by 1994. While advances in combined modality therapy have led to significant progress against locally advanced disease, it was only a decade ago that few believed that the treatment of stage IV non-small-cell lung cancer was justifiable. However, multiple randomized trials in the 1980s and 1990s have changed the role of chemotherapy in lung cancer, such that by the middle of the next decade, it may be that only patients with stage IA non-small-cell lung cancer are not considered as candidates for chemotherapy. PMID- 11484948 TI - The type III epidermal growth factor receptor mutation. Biological significance and potential target for anti-cancer therapy. AB - Mutations in the epidermal growth factor receptor occur frequently in a number of human tumours including gliomas, non-small-cell lung carcinomas, ovarian carcinomas and prostate carcinomas. The type III epidermal growth factor receptor mutation (variously named EGFRvIII, de2-7 EGFR or AEGFR), which lacks a portion of the extracellular ligand binding domain, is the most common. Here, we review the current status with regard to the role of EGFRvIII in human cancers. A detailed discussion of the formation of EGFRvIII and its structure at the protein level are likewise included along with a discussion of its more functional roles. The design and use (preclinical and clinical) of small molecule inhibitors, antibodies, and antisense oligonucleotides against wild-type EGFR are considered in detail as these strategies can be directly adapted to target EGFRvIII. Finally, the status of EGFRvIII targeted therapy is reviewed. PMID- 11484949 TI - Smoking and prostate cancer: findings from an Australian case-control study. AB - BACKGROUND: To examine the risk of smoking on histopathologically-confirmed moderate- and high-grade prostate cancer. MATERIALS AND METHODS: A population based case-control study was conducted in Melbourne, Sydney and Perth between 1994 and 1998 in men aged below 70 years. Cases were recruited from cancer registries and controls were selected from electoral registers. 1498 cases and 1434 controls were interviewed and a detailed smoking history obtained. Data were analyzed by unconditional logistic regression, adjusting for age, study center, year of recruitment, family history and country of birth. RESULTS: The odds ratios (OR) were 1.02 (0.85-1.22) for former smoking and 0.82 (0.65-1.05) for current smoking. The respective ORs were 0.95 (0.78-1.15) and 0.76 (0.59-0.99) for moderate grade tumors, and 1.28 (0.96-1.70) and 1.00 (0.67-1.47) for high grade tumors (P = 0.2 for test that ORs of the two grades were identical). There was no evidence of a dose-response effect for duration of smoking, amount smoked daily, pack-years of smoking and years since quitting and most ORs for these variables were close to unity. CONCLUSIONS: Smoking was not associated with the incidence of prostate cancer. The widths and upper limits of the confidence intervals for the effects of current and former smoking were consistent with weak effects at most. PMID- 11484950 TI - Cisplatin and vinorelbine in advanced and/or metastatic adenocarcinoma of the endometrium: a new highly active chemotherapeutic regimen. AB - PURPOSE: To date the systemic treatment of recurrent and/or metastatic adenocarcinoma of the endometrium (EAC), using both chemotherapy and hormonotherapy (HT), is far from satisfactory. The significant activity of vinorelbine (VNR), a relatively new semisynthetic vinca alkaloid, demonstrated in advanced breast cancer, bronchial adenocarcinoma, and in head and neck cancer, prompted us to carry out a phase II trial employing the combination of cisplatin and VNR in a pluri-institutional series of patients with recurrent and/or metastatic EAC. PATIENTS AND METHODS: Thirty-five patients affected by recurrent and/or metastatic EAC have been treated with CDDP 80 mg/m2 on day 1 plus VNR 25 mg/m2 i.v. bolus on days 1 + 8. This cycle was repeated every 21 days. After three cycles patients were restaged for objective response. Analysis of response rate and duration, overall survival, and toxicity pattern were the main aims of the study. RESULTS: Twenty out of thirty-five patients achieved a major objective response for an overall response rate of 57% (95% confidence limits (CL): 39% 74%). Four patients had a complete response (11%; 95% CL: 3%-27%) with a median progression-free survival (PFS) of eight hundred fourteen days, while sixteen patients had a partial response (46%; 95% CL: 29%-63%) with a median PFS of one hundred eighty-four days. Six patients had stable disease and nine progressed. All patients who achieved a clinical complete response had only a single site of disease at entry, but no association was noted between number of involved sites and likehood of achieving PR. Median overall survival was 240 days, while that of patients with complete and partial response was 855 and 300 days, respectively. Treatment was quite well tolerated with few cases of grade 3-4 myelosuppression. Alopecia was virtually absent and neurotoxicity was mild. One patient complained of an acute pain syndrome at the tumor site. CONCLUSIONS: The CDDP + VNR regimen is quite active against recurrent and/or metastatic endometrial adenocarcinoma, at least in terms of objective response rate which is among the highest ever reported in medical literature. However. duration of objective response and median overall survival are in the disappointing range reported for other regimens. In our opinion the CDDP plus VNR regimen is good enough to be compared to the anthracycline-based regimens and may represent the basis for future development of newer active polychemotherapeutic schedules. PMID- 11484951 TI - A retrospective analysis of the relationship between changes in serum PSA, palliative response and survival following systemic treatment in a Canadian randomized trial for symptomatic hormone-refractory prostate cancer. AB - BACKGROUND: To investigate the relationship between changes in serum PSA, palliative response and survival following systemic treatment for symptomatic hormone-refractory prostate cancer (HRPC). PATIENTS AND METHODS: A retrospective review of 161 patients, treated with mitoxantrone and prednisone (M + P) (n = 80), or prednisone alone (P) (n = 81) from a Canadian randomized phase III clinical trial. PSA response was defined by > or =50% decline compared to baseline. Palliative response was defined by the primary and secondary endpoints of the trial. All responses were required to be maintained on two visits at least three weeks apart. The Cox proportional hazards model and a landmark analysis (at nine weeks) were used to evaluate survival differences between PSA responders and non-responders. RESULTS: Using an intent-to-treat analysis in which patients with missing PSA data are considered non-responders, 34% of M + P and 11% of P patients achieved a PSA response (P = 0.0001). Nineteen of thirty-six (53%) patients with PSA response and twenty-six of ninety (29%) patients without PSA response achieved a palliative response (P = 0.001 Chi-square test, phi coefficient = 0.28). From the landmark analysis. PSA responders had longer survival than non-responders (P = 0.009). In multivariate analysis, better performance status, higher hemoglobin and PSA response (P < 0.001) predicted for survival, but palliative response did not (P = 0.11). CONCLUSIONS: There is significant but imperfect statistical association between PSA response and palliative response. PSA response was associated with longer survival. Patients treated with M + P were more likely to achieve a PSA response and a palliative response than those treated with P. PMID- 11484952 TI - Rb, mcl-1 and p53 expression correlate with clinical outcome in patients with liver metastases from colorectal cancer. AB - BACKGROUND: Thymidylate synthase (TS) has been associated with clinical outcome in disseminated colorectal cancer. However, many patients with low TS expression still fail to respond to treatment. Therefore, we studied the cell cycle proteins, Rb, E2F2, Ki67, p21 and p53 and the apoptotic proteins, mcl-1, hax, bcl xl, bcl-2, Fas receptor, Fas ligand, caspase-3, M30 and PARP as potential predictive factors. PATIENTS AND METHODS: In biopsy specimens of liver metastases from 31 colorectal cancer patients, protein expression was retrospectively determined by immunohistochemistry and related to response to hepatic arterial or intravenous (i.v.) 5-fluorouracil (5-FU) treatment, time to tumour progression (TTP) and overall survival. RESULTS: Expression of both p53 and Rb correlated with survival benefit after 5-FU treatment. A median survival time of 79 weeks was found in patients with high levels of p53 or Rb compared to 36 and 44 weeks for patients expressing low levels of p53 (P = 0.027) or Rb (P = 0.030), respectively. Multivariate analysis showed that p53 was the best predictor of survival independent of sex, age or prior treatment. Following 5-FU hepatic arterial infusion, patients with a high TS expression had a shorter survival time than those with a low expression (P = 0.025). The anti-apoptotic protein mcl-1 was the only factor, which correlated with response to 5-FU treatment. Thirty five percent of patients with a diffuse mcl-1 expression responded whereas ninety percent of patients with a peri-nuclear expression responded (P = 0.041). CONCLUSIONS: These results indicate that besides TS, also Rb, p53 and mcl-1 are correlated with clinical outcome in patients with liver metastases from colorectal cancer. PMID- 11484953 TI - Dose-finding and pharmacological study of ifosfamide in combination with paclitaxel and carboplatin in resistant small-cell lung cancer. AB - BACKGROUND: To find the maximum tolerated dose for ifosfamide in combination with paclitaxel and carboplatin in small-cell lung cancer patients (SCLC), who are resistant to cyclophosphamide, doxorubicin and etoposide (CDE). PATIENTS AND METHODS: Different dose schedules of ifosfamide were combined with fixed doses of paclitaxel 175 mg/m2 and carboplatin AUC 6 mg/ml min. Included were 30 patients, with a median age of 60 years, and median time off prior cytotoxic treatment of 8 weeks. All patients were previously treated with CDE and 11 had received re induction CDE. RESULTS: Dose limiting toxicity of our schedule was persistent thrombocytopenia. None of the patients developed neutropenic fever. Non haematological toxicity was mild, although two treatment-related deaths occurred. Fifty-four percent of patients had a partial response and median survival time was twenty-five weeks. CONCLUSIONS: The maximum tolerated dose of this combination for patients with resistant SCLC is ifosfamide 2000 mg/m2 in combination with paclitaxel 175 mg/m2 and carboplatin AUC 6 mg/ml min administered on the first day of a 21-day cycle. PMID- 11484954 TI - Docetaxel in combination with mitoxantrone and granulocyte colony-stimulating factor as front-line chemotherapy in metastatic breast cancer: a multicenter phase II study. AB - PURPOSE: To evaluate the activity and tolerance of docetaxel in combination with mitoxantrone and granulocyte colony-stimulating factor (G-CSF) as front-line treatment in patients with metastatic breast cancer (MBC). PATIENTS AND METHODS: Fifty-four previously untreated patients with MBC who had bidimensionally measurable disease were enrolled onto the study. Forty-eight (89%) patients had visceral metastases and nineteen (36%) had relapsed within twelve months following adjuvant chemotherapy. Docetaxel (100 mg/m2) was given on day 1 after appropriate premedication and mitoxantrone (20 mg/m2) on day 8. G-CSF (150 mcg/m2/d s.c.) was administered from day 2 to day 6 and from day 9 to day 15. The regimen was repeated every three weeks, on an outpatient basis. RESULTS: In an intention-to-treat analysis, 9 (17%) CRs, 24 (44%) PRs, (overall response rate 61%; 95% confidence interval (CI): 48.1%-74.1%), 12 (22%) SD and 9 (17%) PD were observed. The median duration of response and the median time to tumor progression was 12.5 and 14 months, respectively. The overall median survival was 16.5 months, whilst the probability for one- and three-year survival was 61% and 35%, respectively. Grade 3-4 neutropenia occurred in 37 (69%) patients, and febrile neutropenia in 16 (30%); there was one death due to sepsis. Grade 3-4 thrombocytopenia occurred in four (8%) patients. Grade 2-3 neurosensory toxicity was observed in 8 (15%) patients and grade 2-3 asthenia in 24 (45%). CONCLUSIONS: Docetaxel in combination with mitoxantrone and G-CSF support is an intensified and active front-line regimen for patients with MBC; despite its hematological toxicity, this regimen merits further comparison with other standard anthracycline- and/or taxane-based combinations. PMID- 11484955 TI - Assessment of IAP (inhibitor of apoptosis) proteins as predictors of response to chemotherapy in advanced non-small-cell lung cancer patients. AB - BACKGROUND: Expression of inhibitor of apoptosis family proteins (IAPs) has been shown in vitro to decrease chemosensitivity through caspase inhibition. However, the role of IAPs as predictors of response to chemotherapy in cancer patients remains to be determined. PATIENTS AND METHODS: Using immunohistochemistry, we assessed the expression of the IAP proteins c-IAP1, c-IAP2, and XIAP on tumors from 55 patients with advanced non-small-cell lung cancer (NSCLC) treated with chemotherapy, and correlated that with the observed response to chemotherapy, time to progression and overall survival. RESULTS: Differences were observed in the pattern of staining among the IAP proteins. The expression of c-IAP2 and XIAP was exclusively cytoplasmic. whereas c-IAP1 also displayed nuclear staining. The median expression of tumor cells for c-IAP1, c-IAP2, and XIAP was 70%, 45%, and 25%, respectively, and a correlation was observed between c-IAP1 and c-IAP2 (P = 0.004), and c-IAP1 and XIAP expression (P = 0.013). However, no association was seen between the expression of these proteins and sex, age, tumor size, stage, histology and grade of differentiation. Interestingly, expression of c-IAP1, c IAP2, and XIAP did not predict response to chemotherapy. In addition, the expression of IAPs had no impact on the time to progression or overall survival of this group of patients. CONCLUSIONS: Our results indicate that: 1) there are differences in the level of expression and in the subcellular distribution of c IAP1, c-IAP2, and XIAP in tumors derived from NSCLC patients. 2) The expression of c-IAP1, c-IAP2 and XIAP does not predict the response to chemotherapy in patients with advanced NSCLC. 3) The relation between expression of IAPs and chemosensitivity in cancer patients may be more complex than anticipated by in vitro data. PMID- 11484956 TI - Indium-111 octreotide scintigraphy of Merkel cell carcinomas and their metastases. AB - BACKGROUND: Somatostatin receptor scintigraphy (SRS) may be of interest for staging Merkel Cell Carcinoma (MCC). This study was undertaken to evaluate the sensitivity and specificity of SRS and to determine its role compared to conventional investigations. PATIENTS AND METHODS: From 1993 to December 2000, 20 patients (10 females and 10 males, aged from 38 to 88, mean 66 years) were included prospectively. At the time of SRS: 12 patients had been diagnosed as having stage I disease, 6 stage II and 4 stage III. Two patients had two SRS studies during the course of their disease. SRS was performed with Indium-111 pentetreotide (Octreoscan), a radiolabelled somatostatin analogue. Patients were treated according to the clinical stage. A regular follow-up was scheduled every three months. RESULTS: SRS depicted stage I and II MCC tumour sites with an overall sensitivity of 78% (95% confidence interval (CI): 40%-97%) and a specificity of 96% (81%-100%). The histopathological diagnosis was used as the gold standard. Sites visualised by SRS were compared to those detected with conventional modalities and to follow-up data for all stages: SRS visualised four out of five primary tumour sites, six out of eight lymph node sites, no skin metastases (14 sites in 2 patients), two out of three thoracic metastases and zero out of two hepatic metastases. SRS did not influence treatment decision making in any of the cases. CONCLUSIONS: Although SRS seems highly specific in MCC and could be of help in difficult cases, it cannot be recommended for routine evaluation. PMID- 11484957 TI - Familial microsatellite-stable non-polyposis colorectal cancer: incidence and characteristics in a clinic-based population. AB - BACKGROUND: About 15%-20% of colorectal cancers (CRCs) are familial. While a fraction of these arise in the context of hereditary syndromes, the causes underlying the majority of familial CRCs are not yet understood. PATIENTS AND METHODS: Family history of cancer, clinical characteristics, and microsatellite instability (MSI) in a series of 100 consecutive CRC patients were evaluated. RESULTS: Eighteen patients had a positive family history of CRC in a first-degree relative. Of these, two had a clinical diagnosis of familial adenomatous polyposis (FAP), and three were diagnosed with hereditary non-polyposis colorectal cancer (HNPCC) following results of MSI analysis. A diagnosis of HNPCC was also established in a fourth patient with early onset CRC, who had a second degree relative with CRC, and whose tumor was positive for MSI. The remaining 13 familial CRCs did not show MSI in tumor DNA. The mean age at tumor diagnosis in patients with familial microsatellite-stable (MSS) CRC was higher than in HNPCC and FAP patients and similar to that recorded in sporadic cases. The incidence of second primary malignancies was significantly higher in familial MSS CRC probands (n = 4) compared to patients who did not have a diagnosis of FAP or HNPCC and did not have first-degree relatives affected with CRC (n = 6, in a total of 81 probands with these characteristics). CONCLUSIONS: These results define the existence of a subset of familial CRCs characterized by relatively late age at onset, high incidence of second primary tumors, and absence of MSI in tumor DNA. PMID- 11484958 TI - Estrogen dependent expression of the receptor tyrosine kinase axl in normal and malignant human breast. AB - BACKGROUND: Axl, a member of a family of receptor tyrosine kinases characterized by an extracellular domain resembling cell adhesion molecules and an intracellular conserved tyrosine kinase domain has been reported to induce cell proliferation and transformation. In mice, axl is expressed in the normal mammary gland and over-expressed in aggressive mammary tumors. PATIENTS AND METHODS: We have investigated the expression of axl immunohistochemically in 23 normal human breast samples and in 111 consecutive breast carcinomas. Expression of axl was correlated with tumour characteristics (lymph node involvement, stage, grade) and immunohistochemical expression of ER, PR, Ki-67 and c-erbB-2. RESULTS: In normal tissue, axl localizes to the membrane of breast epithelial cells. Axl protein shows membrane associated staining in high correlation (P = 0.004) with the expression of the estrogen receptor (ER). Axl expression was found in a subset of breast carcinomas and was also correlated with high significance (P < 0.0001) with the presence of ER. CONCLUSION: Our results suggest that axl may serve as a mediator of estrogen stimulation preventing the completion of the breast epithelial life cycle and that estrogen induced axl expression may give a survival signal to cancerous cells, preventing them from dying through apoptosis. PMID- 11484959 TI - Positron emission tomography (PET) with 18F-fluorodeoxyglucose (18F-FDG) for the staging of low-grade non-Hodgkin's lymphoma (NHL). AB - BACKGROUND: Although PET has been shown to be highly sensitive in the primary staging of lymphoma, previous studies with small numbers of patients indicated that low-grade NHL may not always be adequately detected by PET. We undertook this study to determine factors influencing the detection of lesions by PET in low-grade NHL and to evaluate the utility of PET in this indication. PATIENTS AND METHODS: Forty-two patients underwent conventional staging procedures (clinical examination, oto-rhino-laryngologic examination, computed tomography of the chest, abdomen and pelvis, gastroscopy and bone marrow biopsy as well as whole body non-attenuation corrected 18F-FDG-PET RESULTS: PET detected 40% more abnormal lymph node areas than conventional staging in follicular lymphoma but was inappropriate for the staging of small lymphocytic lymphoma where it detected less than 58% of abnormal lymph node areas. PET showed more lesions than conventional staging for peripheral (34% more lymph node areas detected) and thoracic lymph node (39% more) areas but not for abdominal or pelvic lymph nodes (26% fewer areas detected). The sensitivity to detect bone marrow infiltration was unacceptably low for PET. In contrast, PET was as effective as standard procedures for the detection of other extranodal localizations, although a few localizations were detected only by PET and a few others only by conventional procedures. CONCLUSIONS: PET may contribute to the management of patients with low-grade follicular NHL. For the other low-grade lymphoma subtypes, the role of PET is less evident. Further studies using PET to evaluate the results of treatment or to diagnose disease recurrence are warranted in low-grade follicular NHL. PMID- 11484960 TI - Clinical and pathological features of posttransplant lymphoproliferative disorders: influence on survival and response to treatment. AB - PURPOSE: To review the clinical and pathological characteristics of patients with posttransplant lymphoproliferative disorders (PTLD) occurring after solid organ transplantation and determine the influence of these characteristics on response to treatment and survival. PATIENTS AND METHODS: Retrospective review of 32 patients. RESULTS: Overall five-year survival was 59%. Forty-five percent of patients diagnosed within the first year after transplant had advanced disease. Characteristics that were associated with poorer survival were diagnosis within the first year posttransplant, monoclonal tumors and presentation with an infectious mononucleosis-like syndrome. Six of eight patients treated with surgery are alive and disease-free. CONCLUSION: Patients with PTLD can achieve long-term survival. Surgery can play an important role in selected patients. Characteristics that may be associated with poorer survival are diagnosis within the first year after transplant, presence of a monoclonal tumor or an infectious mononucleosis-like presentation. PMID- 11484961 TI - Gemcitabine plus etoposide in chemonaive extensive disease small-cell lung cancer: a multi-centre phase II study. AB - BACKGROUND: Both gemcitabine and etoposide are active in the treatment of small cell lung cancer (SCLC), and are characterised by mild toxicity profiles. The combination of both drugs was found to be feasible and active in a phase I dose finding study in solid tumours. Therefore, a phase II trial was initiated to examine the activity and toxicity of this schedule in extensive disease SCLC. PATIENTS AND METHODS: Forty-two chemo-naive extensive disease SCLC patients were enrolled to receive gemcitabine 1000 mg/m2, days 1, 8 and 15, and etoposide 80 mg/m2, days 8, 9 and 10 of a 28-day cycle. RESULTS: Thirty-seven patients were evaluable for efficacy (five received less than one cycle). No complete responses were observed, but partial responses were seen in 17 patients, yielding an overall response rate of 46%. The median duration of response was 5.8 months. Disease stabilisation was obtained in another 10 patients (27%). The median survival of the 37 protocol-qualified patients was 10.5 months (95% confidence interval (CI): 7.5-12.0). The levels of WHO grade 3 and 4 toxicities were low and clinically manageable. CONCLUSION: In comparison with standard platinum-based regimens, this combination of gemcitabine and etoposide resulted in a somewhat lower response rate, but a similar median survival time. Haematological toxicity was more pronounced than expected from the toxicity data of each agent individually. However, because of its mild non-haematological toxicity, and its ability to be administered in an outpatient setting, this combination provides a reasonable palliative option for patients with extensive disease SCLC. PMID- 11484962 TI - Epidermal growth factor receptor expression in 780 breast cancer patients: a reappraisal of the prognostic value based on an eight-year median follow-up. AB - PURPOSE: Because new therapeutic approaches target tumors expressing epidermal growth factor receptor (EGFR), the aim was to undertake a thorough analysis of the expression profile of EGFR in breast cancer and to reassess its prognostic value. PATIENTS AND METHODS: Tumor EGFR levels were determined by a specific ligand binding assay in 780 consecutive breast cancer patients followed in our institute between 1980 and 1993. Mean age was 61 years (25-85 years). All patients had undergone tumor resection with axillary lymph node dissection: 373 patients (47.8%) underwent mastectomy, 37 (5%) subcutaneous mastectomy and 370 (47.2%) tumorectomy. RESULTS: EGFR levels ranged between non-detectable up to 789 fmol/mg protein. EGFR median value was 9 fmol/mg protein and only a small proportion of patients exhibited a relatively marked EGFR expression. There was no link between tumor size, grade, node status and EGFR tumoral levels. There was a constant and significant decrease in EGFR tumoral levels according to patient age. A significant inverse relationship was found between estradiol receptors (ER) and EGFR. Median follow-up was 97 months with a minimum at 4 months and a maximum at 228 months. From univariate analysis it was found that histological grade, tumor size, node status and ER status were all significant predictors of survival, considering metastasis-free as well as overall survival. Using multivariable analysis, only histological grade, tumor size and node status remained independent predictors of survival. CONCLUSION: EGFR determination is of limited value as a prognostic indicator in breast cancer. PMID- 11484963 TI - Phase II study of vinorelbine in patients with androgen-independent prostate cancer. AB - PURPOSE: To evaluate the efficacy and toxicity of vinorelbine in a phase II study in patients with progressive metastatic androgen-independent prostate cancer. PATIENTS AND METHODS: Forty-seven men with progressive metastatic prostate cancer refractory to first-line or second-line hormonal therapy were treated with vinorelbine, a semisynthetic vinca-alkaloid. Vinorelbine was given, on an outpatient schedule, at 25 mg/m2 weekly for at least eight weeks or until progression or excessive toxicity. RESULTS: Forty-seven patients were included in the study, 33 being evaluable for tumour response, 36 for response to PSA, 21 for clinical benefit and 45 for toxicity. Median actual weekly dose was 19 mg/m2 (range 12.0-26.2 mg/m2). Six of thirty-six patients (17%) demonstrated a biologic response with a 50% or more decline in serum PSA on two consecutive measurements taken at least two weeks apart. The median duration of biologic response was 2.7 months. Two of three patients with measurable disease obtained an objective response but remained unconfirmed. No change disease was reported in 23 patients (49%). On entry into the study, 30 patients had symptomatic bone pain and required narcotic or non-narcotic analgesics. Clinical benefit from vinorelbine was achieved in 15 patients out of 21 (32% of the intent to treat analysis population and 71% of the assessable patients). Due to the low number of questionnaires (QLQ-C30) filled in, it was insufficient to allow any statistical analysis. The median survival was 10.2 months. Toxicity was mainly haematologic with 51% of patients experiencing grade 3 or 4 granulocytopenia. Three patients developed deep vein thrombosis. Non-haematologic toxicity, mainly nausea and neurotoxicity, was mild. CONCLUSION: The administration of weekly vinorelbine appears to be a safe treatment for those patients with androgen-independent prostate cancer and poor prognosis features who require chemotherapy. These results provide data for future investigation of vinorelbine in combination regimens. PMID- 11484965 TI - Chondrosarcoma of bone: an oncological and functional follow-up study. AB - We retrospectively analysed the course of 42 out of 45 patients suffering from different chondrosarcomas which were treated surgically. We found a prospective 5 and 10-year survival of 64% for both time intervals. Follow-up examination was possible in 21 of 45 patients. Most of them were staged as NED, one as AWD, and 8 of 45 were lost to follow-up. From 16 dead patients 12 died of the disease and 4 of unknown but not to tumor related reasons. Survival was depending significantly on the histopathological grade, and the stage according to Enneking's surgical staging system. Eleven out of forty-five patients developed metastases within a mean period of twelve months after surgery. The survival of these patients was significantly reduced. Patients with centrally located tumors exhibited a distinct but non-significant worse survival than those suffering from peripheral tumors. In 18% (8 of 45) recurrence of the tumor was evident within a mean period of 24 months (5-85). Functional evaluation was performed in 21 out of 28 alive patients. After a mean follow-up time of 72 months the mean score was 64% (23 100). Females exhibited a distinct but non-significant better result, the same was observed for peripheral locations compared to centrally located tumors. Regarding age, grade, and line of resection no tendency of any dependence was detected, but worst results were seen in those with a stage III tumor and dedifferentiated CS. PMID- 11484964 TI - Among diffuse large B-cell lymphomas, T-cell-rich/histiocyte-rich BCL and CD30+ anaplastic B-cell subtypes exhibit distinct clinical features. AB - BACKGROUND: The EORTC clinical trial 20901, activated in 1990, was designed to treat non-Hodgkin's lymphomas (NHL) of intermediate/high-grade malignancy according to the Working Formulation. Established in 1994, the R.E.A.L. Classification on NHL has now replaced all former classifications. PATIENTS AND METHODS: We reanalysed all cases (n = 273) documented by material available for review according to the R.E.A.L. Classification. In addition, we subdivided cases recognised as diffuse large B-cell lymphoma (DLBCL) into three morphologically distinct categories, namely, large cleaved DLBCL (LC-DLBCL), T-cell rich/histiocyte-rich B-cell lymphoma (T-cell-rich/histiocyte-rich BCL) and CD30+ DLBCL with anaplastic cell features (CD30+ DLBCL). Finally, T/NULL anaplastic large-cell lymphoma (ALCL) cases were subdivided into ALK+ and ALK- lymphomas. Review was performed independently by two pathologists from two different centres. RESULTS: DLBCL (61%), T/NULL ALCL (15%) and mantle-cell lymphoma (MCL, 50%) were the main NHL categories represented in the study. Fifty-seven of one hundred sixty DLBCL cases were further subclassified as LC-DLBCL (33 cases), T cell-rich/histiocyte-rich BCL (13 cases) or CD30+ DLBCL (11 cases). The remaining cases were indicated as unspecified DLBCL. A clinico-pathological correlation confirmed the findings of previous studies suggesting that MCL, DLBCL and ALCL represent distinct entities with MCL being characterised by a short survival, in contrast with the longer survival and less frequent progression typical of ALK+ compared to ALK- ALCL. Within DLBCL, T-cell-rich/histiocyte-rich BCL showed distinctive features at presentation whereas CD30+ DLBCL showed a trend towards a more favourable prognosis, that might be comparable to that of ALK+ ALCL. CONCLUSIONS: Our data further support the usefulness of the R.E.A.L. Classification and illustrate the feasibility of DLBCL subtyping. Moreover, our results demonstrate the distinct clinical characteristics of T-cell rich/histiocyte-rich BCL and CD30+ DLBCL with anaplastic cell features suggesting that they may represent clinico-pathologic entities. PMID- 11484966 TI - Pancreatic metastasis of alveolar soft-part sarcoma: a case report and review of the literature. AB - Alveolar soft-part sarcoma is a rare tumour. Patients commonly present with distant metastases both at the time of diagnosis and late in the course of disease. We present a case of pancreatic metastasis, occurring more than six years after diagnosis. Treatment consisted in subtotal pancreatoduodenectomy with pylorus resection. Both specific patterns of relapse and treatment opportunities of this uncommon feature are discussed. PMID- 11484967 TI - Gastric linitis adenocarcinoma and carcinomatous meningitis: an infrequent but aggressive association--report of four cases. AB - Carcinomatous meningitis (CM) is a very rare complication of gastrointestinal malignancies and especially gastric adenocarcinoma. Linitis plastica (LP), which is a specific form of gastric neoplasia, locally penetrates through the gastric wall to reach the peritoneum. Lymph node involvement is frequent and metastatic sites are almost exclusively observed in the abdominal cavity. The meningeal localization is extremely rare with only a few cases described in the literature. We report here, over a five-year period, four cases of CM on a total of eighty linitis cases diagnosed and treated in our institution, which represent 5% of a non selected linitis population. The clinical manifestations were clearly poor, and characterized by aspecific neurological signs. The diagnosis was made by the discovery of signet cells in the cerebrospinal fluid. Invasive treatment, consisting of intrathecal infusion of chemotherapy, was undertaken with mixed clinical response and no cytological normalization of the cerebrospinal fluid (CSF). In conclusion, our observation which is based on a large series of successive gastric linitis, demonstrates a 5% frequency of developing CM with a predominance among metastatic patients. Furthermore, the diagnosis of CM must be done as soon as possible because of the clear effectiveness of a therapeutic approach on the improvement of symptoms and quality of life. PMID- 11484968 TI - Necrosis in melanoma metastases with treatment. PMID- 11484969 TI - Reaction after oxaliplatin--prevention with corticosteroids? PMID- 11484970 TI - Cost of de novo acute myeloid leukemia induction therapy in adults: analysis of EORTC-GIMEMA AML10 and FLANG regimens. AB - Since the social and financial impact of AML therapy is becoming more and more relevant we analyzed the cost of induction therapy of two different regimens. The first one is part of the widely employed EORTC-GIMEMA AML-10 and consists often days of therapy. The second (FLANG) is a short (three day), Fludarabine, Ara-C, mitoxantrone and G-CSF containing regimen. We first retrospectively analyzed the outcome of 77 consecutive AML patients with comparable clinical and haematological features receiving FLANG (25) or AML-10 (52), between June 1993 and October 1999, and observed equivalent CR rate, as well as DFS and overall survival duration. We then selected 9 non pretreated patients per group who reached CR after one course of therapy. Patients treated with FLANG had a statistically significant earlier platelet recovery compared to those treated with AML-10, fewer days of intravenous antibiotic therapy (14/22, respectively, p < 0.05), and a shorter hospitalization period (22/33 days, p < 0.01). FLANG was significantly more expensive than AML 10 as far as the cost of antiblastic drugs (p < 0.01) and G-CSF support (p < 0.05) are concerned. On the contrary, the expense for antiemetic drugs (p < 0.01) and the cost of personnel and other services ($5,906/$3,970, p < 0.05) were higher for AML-10 than for FLANG. Overall, the average costs of FLANG and AML10 were $9,269 and $12,424 respectively (p < 0.05; difference = -25%). Our study seems to indicate that, compared to AML-10, FLANG induction is as effective, less expensive and it allows for a decrease in the length of hospitalization and thus for better exploitation of the financial resources of Hematology-Oncology departments. PMID- 11484971 TI - Human dendritic cells pulsed with autologous Epstein-Barr virus transformed B cell lymphoblastoid cell (BCL) lysate elicit a BCL specific MHC-class II restricted T-cell response. AB - Epstein Barr Virus (EBV) associated lymphoproliferative disorders (LPD) express EBV latent antigens that are also expressed on normal B-cells transformed with EBV. This could potentially be exploited to develop immunotherapeutic strategies for LPD and other EBV associated malignancies. To this end we investigated the capacity of human monocyte derived dendritic cells (DC) pulsed with lysate from autologous EBV transformed B-cell lymphoblastoid cell (BCL) lysate to elicit an in vitro antitumor response. BCL lysate pulsed DC generate BCL specific cytotoxic lymphocytes, as lymphocytes primed with such DCs induce cytolysis of autologous (>60%) but not allogeneic BCL (<5%). In addition, lymphocytes primed with BCL lysate pulsed DC secrete gamma-IFN (3176 pg/ml). Whereas gamma-IFN production was markedly reduced (>99%) when BCL specific T-cells were stimulated by BCL lysate pulsed DC in the presence of blocking antibodies to HLA-DR, DP and DQ, use of antibodies to MHC class-I resulted in only a minimal reduction in gamma-IFN production (17%). These studies demonstrate that BCL lysate pulsed DC elicit a predominantly BCL specific, MHC class-Il restricted T cell response. This suggests that vaccination with autologous BCL lysate pulsed DC may represent a viable immunotherapeutic approach for the treatment of LPD. PMID- 11484973 TI - Unusual salivary glands adenocarcinomas: a retrospective study. AB - Malignant tumors of the salivary gland ducts exhibit variable morphological patterns and biological behavior. This group of neoplasms includes some uncommon lesions that originate from the intercalated ducts, such as polymorphous low grade adenocarcinoma, epithelial-myoepithelial carcinoma and basal cell adenocarcinoma, and other that arise from the excretory duct, such as the salivary duct carcinoma. One hundred and seventy-seven malignant neoplasms of the major and minor salivary glands were reviewed. There were 23 cases of uncommon types of ductal carcinomas. Age, sex, location, size of tumor were examined as well as their histopathological features. There were four polymorphous low-grade adenocarcinomas (17.3%), two of which were located on the palate and two on the cheek; seven epithelial-myoepithelial carcinomas (30.4%), most of them located in the parotid; six basal cell adenocarcinomas (26%) which predominated in the submandibular gland, and there were six salivary duct carcinomas (26%), four of which occurred in the parotid gland, and two in the submandibular gland. PMID- 11484972 TI - Hepatic arterial infusion of cisplatin diluted in hypotonic 25 g/l glucose solution administered in balloon-occluded hepatic artery: experimental rationale and clinical pilot study. AB - Reduced osmolarity in incubation medium was previously shown to increase in vitro cellular accumulation and cytotoxicity of cisplatin on cancer cells. We confirmed in the present work that cisplatin diluted in an hypotonic 25 g/l glucose solution (124 mOsm) was dramatically more cytotoxic in vitro than cisplatin diluted in normotonic 9 g/l NaCl (300 mOsm) on the human HT29 colon and MCF7 breast cancer cells. We conducted then a pilot clinical study on the administration of cisplatin diluted in hypotonic 25 g/l glucose solution given through the balloon-occluded hepatic artery for the treatment of liver metastases from colon or breast cancer tumors. Nine patients (5 men, 4 women; mean age 58, range: 44-71) with confirmed isolated, unresectable metastases from colorectal (7) or breast (2) tumors were included in this study and a total of 23 cycles were administered (2.55 per patient; range 1-5) with an average dose of 50 mg cisplatin (range: 12.5-100). Hepatic artery dissection due to balloon injury with partial or complete arterial obstruction were encountered in 2 patients. Pain in the liver and epigastric area was the main symptom which was constant and intense during the IAH cisplatin injection. Fever > 38 degrees C was observed in 15/23 cycles and increase of creatinine in 1/23 cycles. Transient increase of hepatic transaminases without change in prothrombin time was registered in all patients. However one patient who received the highest dose of 100 mg cisplatin developed a persistent but reversible clinical jaundice and a transient increase in prothrombin time. One patient achieved a partial response (12 weeks), 7 had stable disease (mean duration: 6 weeks) and one had a progressive disease. Hepatic arterial infusion of cisplatin diluted in hypotonic 25 g/l glucose solution and administered through the balloon-occluded hepatic artery is a feasible approach. Total dose of cisplatin in hypotonic glucose solution will not exceed 80 mg by cure in a further phase II study. PMID- 11484974 TI - Gemcitabine in pre-treated advanced renal carcinoma: a feasibility study. AB - Eigtheen patients affected by metastatic renal cell carcinoma, 16 which were assesable, were treated with 1 g/m2 of Gemcitabine (GCB) on days 1, 8 and 15 of a 28-day treatment cycle up to a maximun of ten cycles. All patients in neoplastic progression were treated with chemo- and immunotherapy (5 FU, IL-2, IFN alpha d 13-cis-retinoic acid.) Out of the 16 assessable patients, 5/16 (31%) showed overall response (ICR, 4 PR), 5 (31%) stable disease (SD) and 6 (38%) progression of disease (PD). Toxicity was limited to WHO grades I only, primarily hematological. PMID- 11484975 TI - Analysis of a follow-up program for anal canal carcinoma. AB - The ideal follow-up program for anal canal cancer remains unclear and controversial. We hereby describe an extensive follow-up program for anal canal carcinoma in order to evaluate which examinations and which diagnostic techniques really had impact on survival and management. We evaluated 25 patients with anal canal carcinoma. Local excision (LE) was performed in 5 patients, radiochemotherapy (RCT) in 13, radiochemotherapy and local excision (RCTE) in 7. Mean follow-up time was 6.3 years (range 20 months-11 years). The follow-up program included clinical examination, serum tumor markers evaluation, transrectal ultrasonography (TRUS), anoscopy with either mucosal or by Tru-cut needle multiple biopsies, standard chest X-ray and hepatic-inguinal ultrasonography, endoanal magnetic resonance imaging and in some cases total-body skeletal scintigraphy. A large multicentered randomized and prospective trial is surely lacking and should be undertaken as soon as possible. Our results suggest that an effective local control, rather than a higher survival is the reachable goal at present for anal canal carcinomas. However, further steps should be made to achieve better results. After this experience we propose a more semplified follow-up protocol which consists in performing only rectal examination, endoscopy, Tru-cut needle biopsies and TRUS for local control and inguinal ultrasound and TC to evidence distant metastases. PMID- 11484976 TI - Inhibition of peritoneal dissemination in human gastric cancer by MMP-7-specific antisense oligonucleotide. AB - MMP-7 is a matrix-degrading enzyme that is mainly produced from cancer cells, and has a great role in the invasion and metastasis of cancer. We have established a highly metastatic cell line (MKN-45-P) on the peritoneum of nude mice from MKN-45 by repeated intraperitoneal inoculation of intraperitoneal free cancer cells. By the precise screening of metastasis-related genes using reverse transcriptase polymerase chain reaction (RT-PCR), MKN-45-P characteristically expressed more MMP-7 than the original cell line of MKN-45. In this study, we studied the effects of antisense oligonucleotides complementary to exon 3 of MMP-7 mRNA on the expression of MMP-7 and metastatic potential of MKN-45-P by using in vitro and in vivo experiments. RT-PCR and western blot analysis demonstrated that 10 microM antisense oligonucleotides suppressed MMP-7 expression at both the mRNA level (84%) and protein level (56%). Antisense oligonucleotides, specific for MMP 7 suppressed invasion by MKN-45-P cells without influencing proliferation. On the other hand, scrambling sequence control oligonucleotides did not show any inhibitory effects. In addition, survival of MKN-45-P bearing mice, which had been treated for 48 hrs with antisense oligonucleotides before intraperitoneal injection, was significantly better than that of control mice. In contrast, control oligonucleotides did not influence the survival of mice with the peritoneal dissemination model. These results strongly suggest that MMP-7 may have a great role in the formation of peritoneal dissemination in gastric cancer, and the molecular control of MMP-7 using antisense oligonucleotides may be a hopeful treatment modality for peritoneal dissemination. PMID- 11484977 TI - Effect of a matrix metalloproteinase inhibitor on a lymph node metastatic model of gastric cancer cells passaged by orthotopic implantation. AB - Lymph node metastasis is the most frequent type of tumor recurrence and is known to be one reason for the poor prognosis in patients with digestive cancer. However, the mechanisms of lymph node metastasis are not clearly understood and a metastatic model will be useful for elucidating factors associated with lymph node metastases. In this study, we investigated the effect of R-94138, a matrix metalloproteinase (MMP) inhibitor, on the lymphnodal metastatic ability of gastric cancer cells using an in vivo orthotopic implantation model in nude mice. Injection of a gastric cancer cell line, MKN-45, into the gastric wall resulted in lymph node metastasis 8 weeks after inoculation. The number of lymph node metastases and the amount of body weight loss significantly decreased by intraperitoneal administration of R-94138. Histologically, lymphatic invasion of cancer cells was found in primary gastric tumors of control mice with lymph node metastasis. However, no lymphatic invasion was observed in the gastric wall of R 94138-treated mice without lymph node metastasis. These findings suggested that the MMP inhibitor, R-94138, could be used in adjunctive therapy for lymph node metastasis in gastric carcinoma. PMID- 11484978 TI - Effect of Brahma Rasayana on antioxidant systems and cytokine levels in mice during cyclophosphamide administration. AB - Intraperitoneal administration of cyclophosphamide (CTX) 25 mg/kg.b.wt. dose/mouse for 10 days was found to suppress the tissue and serum level of reduced glutathione (GSH), blood glutathione peroxidase (GPX) and tissue levels of superoxide dismutase (SOD) and catalase (CAT). Tissue levels of glutathione reductase (GR) and glutathione-S-transferase (GST) were unaltered by CTX treatment while serum and tissue lipid peroxide levels were significantly increased. Oral administration of Brahma Rasayana BR-50 mg/dose/mouse for 10 days and 30 days significantly enhanced the tissue levels of SOD, CAT, GST, GPX, serum and tissue GSH and significantly reduced the serum and tissue lipid peroxidation. BR treatment was also found to enhance the serum cytokine level of interferon gamma (IFN-gamma), interleukin-2 (IL-2) and granulocyte macrophage-colony stimulating factor (GM-CSF) in normal and CTX treated mice. The results are indicative of the use of BR to reduce the oxidant stress induced by CTX treatment and its effect in cellular function. PMID- 11484979 TI - Primary tumours neoangiogenesis and P53 expression in oral carcinoma patients. AB - Paraffin embebbed tumour tissues from 47 T1-2 N0-1 M0 primary oral squamous carcinoma have been utilized for immunohistochemical analysis of p53 expression (moab DO-7) and microvessel density (MVD) analysis (moab CD34). Fifty percent of cases showed p53 immunostaining with an average of 21% of p53 positive cells. A strong trend for a longer survival in patients with tumor p53- versus p53+ was evidenced (median survival: 12 months versus not reached, respectively; p=0.08 by log-rank test). A mean value of 27 MVD was found. The probability of overall survival did not result significantly different in the subgroups of tumours with high and low MVD (median survival: 6 months versus 6 months, respectively; p=0.24). Cox multivariate analysis confirmed that the only prognostic factor significantly related to the overall survival was clinical nodal status (O.R.=2.7; 95% C.I. 1.09-6.9), while p53 status only approached the statistical significance (O.R.=2.5; 95% C.I. 0.96-6.5; p=0.06). PMID- 11484980 TI - Membrane proteins in neoplasic and normal uterine cervix. AB - Tumor cells exhibit phenotypic and genotypic differences in comparison to normal cells. These differences can be used to identify proteins important for tumor growth and, therefore, potentially used in the diagnosis and follow-up of patients. The objective of this work was to investigate the electrophoretic pattern of cytoplasm membrane proteins from normal and malignant cervix using polyacrylamide-SDS gels. A highly reproducible protein pattern was found in the 29 samples of normal cervix whereas three well-defined patterns of protein bands were observed in the 48 tumor specimens (pattern I: 25%, pattern II: 29.2% and pattern III: 45.8%). A low concentration or absence of high molecular weight proteins was observed (p<0.5) in tumor samples. None of the tumor protein patterns correlated with the clinicopathologic characteristics of patients. Nine out of 11 patients (82%) showing the pattern III had a complete clinical response whereas only 55% (11 out of 20) of those with patterns I and II showed a complete response. However, this difference was non-significant (p=0.1247). In conclusion, we demonstrate that there is a gain and loss of cytoplasmic membrane proteins in tumors, shown as different protein band patterns. These findings could have clinical and biological significance that must be further evaluated. PMID- 11484981 TI - Expression and prognostic significance of urokinase and plasminogen activator inhibitor type-1 in endometrial hyperplasia and cancer. AB - Proteolytic enzymes, like urokinase (uPA) and plasminogen activator inhibitor type-1 (PAI-1), are involved in remodelling tissues during invasion and metastasis of tumor cells. The purpose of the study is to evaluate the expression and the prognostic significance of these enzymes in endometrial hyperplasia and cancer. We used immunohistochemical staining to localize uPA and PAI-1 antigens and evaluate their expression, and the enzyme-linked immunosorbent assay (ELISA) to measure their levels during the progression of endometrial carcinoma. The results show that the levels of uPA and PAI-1 detection are systematically weak in simplex hyperplasia and are moderate in complex hyperplasia. In the endometrial carcinoma a very strong reaction was observed in the most aggressive variant of epithelial tumors. A positive signal for uPA was found only in the cytoplasm of normal and hyperplastic cells while, in tumors, uPA was present also in the cellular areas surrounding the neoplastic glands and at the apex of the malignant cells. The PAI-1 immunoreactivity was weak to moderate in 95.4% of carcinomas, with a diffuse signal mostly distributed in the cytoplasm of neoplastic cells and tumor stroma. UPA antigen concentrations were significantly higher in endometrial carcinoma than in endometrial hyperplasia (p<0.05) and in normal endometrium (p<0.001). PAI-1 antigen concentrations in carcinoma samples were significantly higher than in normal endometrium (p=0.002), but the difference was not statistically significant with respect to that in endometrial hyperplasia. We did not find any correlation between uPA and PAI-1 concentrations and the standard prognostic parameters for evaluating endometrial carcinoma. In conclusion, this study demonstrates that in hyperplastic endometria and in endometrial carcinoma there is a progressive increase in expression of uPA and PAI-1 than in normal endometrial tissue. In carcinoma tissues, the high expression of uPA is unregulated in the surrounding stroma tissue, particularly in the most aggressive histopathologic variants. UPA and PAI-1 may be factors associated with invasive behavior in endometrial carcinoma independent of other clinicopathological parameters. PMID- 11484982 TI - Plasminogen activator inhibitor 1 (PAI-1) levels and gene promoter polymorphisms in subjects with colorectal cancer. AB - The high level of plasminogen activator inhibitor 1 (PAI-1) in colorectal cancer predicts poor prognosis for patients. The insertion (5G)/deletion (4G) polymorphism (the 4G/5G polymorphism) and G-->A single base substitution (the G/A polymorphism) located at promoter of PAI-1 gene may have functional significance in regulation of its expression. In the present work the level of PAI-1, distribution of genotypes and frequency of alleles of the 4G/5G and G/A polymorphisms in samples of cancer tissue and normal mucosa as well as in blood were investigated. Blood, tumor and normal tissues were obtained from 40 patients with colorectal cancer. The 4G/5G and G/A polymorphism were determined by PCR amplification using the allele specific primers. The PAI-1 level was measured by enzyme linked immunosorbent assay (ELISA). The distribution of the genotypes of both polymorphisms did not differ significantly (p > 0.05) from those predicted by the Hardy-Weinberg distribution. There were no differences in the genotype distributions and allele frequencies between blood, normal mucosa samples and cancer tissue. The 4G/5G and G/A polymorphisms were in linkage disequilibrium. The average level of PAI-1 in tumor samples was significantly (p < 0.05) higher than in normal tissue. The results obtained indicate that a higher level of PAI-1 can be associated with colorectal cancer. On the other hand, in colon cancer, the 4G/5G and G/A polymorphisms are not linked with elevated levels of PAI-1 and therefore may not be used to predict colon cancer prognosis. PMID- 11484983 TI - Serum cytokeratins determination in differentiated thyroid carcinoma. AB - To assess the potential value of cytokeratins (CK) 8,18,19 as tumor markers for thyroid diseases, a study was performed comparing serum CK 8,18,19 levels in patients affected from thyroid carcinoma, adenoma, other benign thyroid diseases and healthy volunteers as controls. One hundred cases (65 patients and 35 controls) were examined. Thirty patients had thyroid carcinoma (18 papillary- PTC, 8 follicular--FTC, 4 medullary--MTC), 19 non-toxic goiter, 10 thyroid adenoma, 6 chronic thyroiditis and 35 healthy volunteers as controls. These controls were matched by age and sex. The mean value of CK in benign thyroid diseases (46.1 U/L) was significantly higher (p<0.02) than that in healthy controls (29.6 U/L). The mean value of CK in carcinomas (68.1 U/L) was significantly higher than that in healthy controls (p<0.01) and benign thyroid diseases patients (p<0.05). The positive rate of CK in thyroid carcinomas was 28.1%, while in benign thyroid diseases was 17.8%. The CK sensitivity for thyroid carcinomas was 28.1%, with a specificity of 80% and accuracy of 70.4%. In PTC patients the mean CK value was not significantly higher than in the benign diseases' group and in healthy subjects. No evident correlation between CK levels and tumor mass was found. In FTC patients the mean value was significantly higher than in the benign diseases' group and in healthy subjects. Large tumors showed the highest levels, while small tumor values were similar to the control ones. In MTC patients the mean value was significantly higher than in the benign diseases' group and in healthy subjects, with the highest peaks in large tumors and metastatic tumors. The detection of increased values in thyroid carcinomas with high metastatic potential (FTC and MTC) seems to confirm the role of these antigens in predicting the malignancy's degree of the neoplasm. These findings, if confirmed in larger series, could play an important role in assessing the CK 8,18,19 serum level as a real prognostic factor. Further repeated serum determinations after total thyroidectomy might indicate the role of CK 8,18,19 as serum markers predicting the risk of metastases. PMID- 11484984 TI - Correlation between spontaneous apoptosis and the expression of angiogenic factors in advanced gastric adenocarcinoma. AB - The aim of this study was to investigate whether angiogenic factors influence the occurrence of spontaneous apoptosis in advanced gastric cancer. The apoptotic indices (AIs) of 97 tumors from 97 patients with advanced gastric cancer (pT3, pN0, pM0, Stage II) were analyzed by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) method. Intratumoral microvessel densities (IMVDs) of tumors stained with anti-CD34 monoclonal antibody were quantified under x 200 magnification using computer assisted image analysis. The expressions of angiogenic factors, such as vascular endothelial growth factor (VEGF), thymidine phosphorylase (dThdPase), transforming growth factor-alpha (TGF-alpha), and p53 were analyzed immunohistochemically and compared with IMVDs and AIs. The mean IMVD of the 97 tumors was 365/mm2 (range 147-990/mm2). The mean AI of tumors was 2.1% (range 0 11.3%). A significant inverse correlation between the AIs and the IMVDs was shown (p = -0.278, P = 0.0064). The mean IMVDs of tumors with high expressions of dThdPase, TGF-alpha, or p53 were significantly higher than those of tumors with low expressions of these factors. The mean AI of tumors with high expressions of dThdPase was significantly lower than that of tumors with low expressions of dThdPase (P = 0.023). However, no significant correlations were detected between AIs and the expression levels of VEGF, TGF-alpha, or p53. In gastric cancer, dThdPase may play an important role in tumor progression by increasing microvessels and by suppressing apoptosis of cancer cells. PMID- 11484985 TI - Tissue transglutaminase expression in breast carcinomas. AB - Tissue transglutaminase (tTG) is known to participate in multiple cellular processes, including apoptosis, cellular adhesiveness etc. Alterations of tTG expression could contribute to the development of several categories of diseases, including AIDS, cancer etc. The aim of the study was to test the pattern and relevance of tTG expression in a subset of breast carcinomas. RT-PCR has detected tTG-specific RNA message in 11 out of 25 (44%) breast cancer samples. tTG message was detected in 6/8 (75%) breast carcinomas with high apoptotic index, but only in 5/17 (29%) with the low one (p = 0.03). Immunohistochemical analysis revealed that only 15% of breast carcinomas displayed tTG protein in tumor cells, while the staining of the stromal components occurred in approximately one-half of the tumours tested. Surprisingly, there was no significant association between tTG RNA expression and protein positivity. Moreover, there was no evident relationships between tTG immunostaining and apoptotic index or clinical parameters of breast neoplasms. There are at least 2 alternative explanations for the poor concordance between RNA and protein data. It is likely that the sensitivity of immunohistochemistry is not sufficient to detect functionally relevant tTG enzyme in all breast cancer sections. Otherwise, tTG RNA expression does not always lead to accumulation of its product in the tumor cells, but reflects the transcriptional activation of other pro-apoptotic genes due to common triggering mechanisms. PMID- 11484986 TI - Studies on the expression of alpha(v)beta3 integrin receptors in non-malignant and malignant human cervical tumor tissues. AB - Integrins are group of cell surface receptors, which mediate adhesion between cell-cell and cell-extracellular matrix proteins of the basement membrane. Integrins play an important role in cellular growth, development, morphology, signalling and also in tumor development. Among the integrin group of cell surface receptors one of the most important member is alpha(v)beta3 integrin receptor. Evidences say that the expression of this integrin receptor is regulated during tumor development. Large numbers of studies have been done to establish the role of alpha(v)beta3 integrin receptor in human melanoma systems. Expression of this receptor in metastatic but not benign melanomas suggests a role for this integrin in the regulation of tumor proliferation. Alpha(v)beta3 integrin receptor also plays a significant role in tumor angiogenesis, apoptosis and signal transduction process. Recent studies show that collagenase MMP-2 binds directly to integrin alpha(v)beta3 on the surface of invasive tumor cells and facilitates tumor cell invasion. In this present communication we studied the expression of alpha(v)beta3 integrin receptor in malignant and non-malignant cervical tumor tissues. Because receptor ligand interaction is a cell surface phenomenon the membrane fraction of tumor tissues was separated and the expression of alpha(v)beta3 integrin receptor was assayed by ELISA and immunoprecipitation from membrane extracted protein fraction. Comparative ELISA and immunoprecipitation clearly demonstrates much higher expression of alpha(v)beta3 vitronectin integrin receptor in the membrane fraction of malignant human cervical tumor tissues than nonmalignant tissue membrane fractions. PMID- 11484987 TI - A pilot study on the relation between cisplatin neuropathy and vitamin E. AB - Peripheral sensory neuropathy is the main non-hematological side-effect related to cisplatin chemotherapy. The strong similarity between clinical and neuropathological aspects in peripheral neuropathy induced by cisplatin and neurologic syndromes due to vitamin E deficiency, prompted us to investigate the relationship between cisplatin neuropathy and plasmatic level of vitamin E (alpha tocopherol). We measured vitamin E in the plasma of 5 patients (Group 1) which developed severe neurotoxicity after cisplatin treatment and in another group of 5 patients (Group 2) we analyzed the plasmatic level of vitamin E before and after 2 or 4 cycles of cisplatin treatment. The results showed that the patients of group 1 presented low plasmatic levels of vitamin E and that the patients of group 2 presented significantly lower levels of vitamin E after 2 or 4 cycles of cisplatin than before treatment. Our preliminary data suggest that an inadequate amount of the antioxidant vitamin E due to cisplatin treatment could be responsible of the peripheral nerve damage induced by free-radicals. Given the lack of toxicity of vitamin E, we need to systematically assess the possible neuroprotective role of vitamin E supplementation in patients treated with cisplatin chemotherapy. PMID- 11484988 TI - Extraocular muscle ultrastructural pathology in the paraneoplastic phenomenon associated with retinoblastoma. AB - Extraocular muscle biopsies were obtained during enucleation because of advanced intraocular retinoblastoma in four patients admitted to the Service of Ophthalmology at the Caracas University Hospital. Slight limitations of ocular movements and strabismus were present in all cases. The electron microscopical analysis showed muscle fibres with slight to severe atrophy exhibiting myopathic structures as nemaline, filamentous and zebra bodies. Fibre necrosis was also observed characterized by sarcomeric hypercontraction, autophagia, sarcolemmal disruption, and mitochondrial swelling. Capillary alterations included endothelial proliferation with intraluminal infoldings and, in some cases, capillary degeneration and necrosis. A mononuclear cell infiltration formed by macrophages and scarce mast cells located next to atrophic fibres and altered capillaries was observed. Additionally, neutrophils were found around capillaries and in their wall. Cancer cells invading muscle tissue were not seen. Two different ethiopathogenic mechanisms for muscle damages seem to be present. Because of the similarity between the microvascular changes we observed and those found in the muscle compromise of several autoimmune diseases, an autoimmune component in the ethiopathogenesis of the observed capillary alterations is proposed. On the other hand, abnormalities observed in muscle fibres are very similar to those in neurogenic atrophy. This study represents the first report on an extraocular muscle paraneoplastic phenomenon associated with orbital tumours. PMID- 11484989 TI - Effect of Caffeine, a xanthine derivative, in the inhibition of experimental lung metastasis induced by B16F10 melanoma cells. AB - Caffeine, a methyl xanthine derivative, was studied to assess the effect on B16F10 melanoma induced experimental metastasis. Caffeine was administered at a dose of 100 and 50 mg/kg body weight by both routes, to tumour bearing animals. Solid tumour reduction studies with Caffeine showed a significant reduction in tumour volume for 100 mg/kg dose by both oral and i.p. routes. The Caffeine treated metastatic tumour bearing animals significantly (p<0.001) inhibited lung tumour nodules. Serum sialic acid levels and lung hydroxyproline contents in the treated groups were significantly (p<0.001) low, when compared with the untreated control animals. In the present study, our results suggest that Caffeine inhibits solid tumour development and pulmonary experimental metastasis induced by B16F10 melanoma cells, in murine model. PMID- 11484990 TI - Adenosquamous carcinoma of the colorectum: report of two cases. AB - Adenosquamous carcinomas of the colorectum are rare neoplasms. Our experience with two cases is presented in this paper. One patient, who complained of bloody stool, was found to have adenocarcinoma in the sigmoid colon. He received a laparoscopy-assisted sigmoidectomy. The histological examination revealed that the tumor was adenosquamous carcinoma. To date, he has survived six months post operatively without evidence of recurrence. The other patient, who complained of anal bleeding, was found to have rectal adenocarcinoma and received a low anterior resection. Histological examination revealed that the tumor was an adenosquamous carcinoma. He remains alive, with no evidence of recurrence, nine years post operatively. Both cases showed paracolic lymph node metastasis. Because of its very low incidence, the histogenesis, malignancy and prognosis of this disease remain unclear. Thus, further clinical and histological study of this disease entity is required. PMID- 11484991 TI - Cutaneous metastases from gastric adenocarcinoma treated with combination chemotherapy producing complete response with long survival. AB - A case of cutaneous metastases from gastric adenocarcinoma was presented. It was treated with combination chemotherapy producing complete response with prolongation of survival. Skin metastasis will indicate early fatal termination, therefore there has not yet been a reported case of cutaneous metastasis from gastric cancer to have complete response with long survival. PMID- 11484992 TI - Rare presentation of carcinosarcoma arising in bladder diverticulum. AB - An 87-year-old man presented with hematuria and dysuria. An endoscopic examination revealed a bladder mucosa which was almost entirely occupied by diverticula. On the left lateral wall of the bladder there was a huge diverticulum which contained a 12 cm mass extending beyond the bladder wall in the extraperitoneal tissues. Upon histological examination the mass proved to be a carcinosarcoma which was composed by a squamous carcinoma and a sarcoma resembling a malignant fibrous histiocytoma. The two components, i.e. carcinomatous and sarcomatous, were separated by a sharp collision border and no intermingling was ever noted. The epithelial component showed immunoreactivity for cytokeratin and EMA, while the mesenchymal component was diffusely reactive for vimentin, alpha-1-antitrypsin and lysozime. Both components were reactive for galectin-3, whereas S100, desmin and smooth muscle actin were negative. This is the fourth reported case of carcinosarcoma originating in a bladder diverticulum. PMID- 11484993 TI - Lack of HLA-DR expression in a patient with B-chronic lymphocytic leukemia. AB - We describe a rare case of a patient with chronic lymphocytic leukemia (CLL) whose immunophenotype analysis revealed lack of HLA-DR in B-cells. PMID- 11484994 TI - Ontogenetic changes in the neural mechanisms of eyeblink conditioning. AB - The rodent eyeblink conditioning paradigm is an ideal model system for examining the relationship between neural maturation and the ontogeny of associative learning. Elucidation of the neural mechanisms underlying the ontogeny of learning is tractable using eyeblink conditioning because the necessary neural circuitry (cerebellum and interconnected brainstem nuclei) underlying the acquisition and retention of the conditioned response (CR) has been identified in adult organisms. Moreover, the cerebellum exhibits substantial postnatal anatomical and physiological maturation in rats. The eyeblink CR emerges developmentally between postnatal day (PND) 17 and 24 in rats. A series of experiments found that the ontogenetic emergence of eyeblink conditioning is related to the development of associative learning and not related to changes in performance. More recent studies have examined the relationship between the development of eyeblink conditioning and the physiological maturation of the cerebellum, a brain structure that is necessary for eyeblink conditioning in adult organisms. Disrupting cerebellar development with lesions or antimitotic treatments impairs the ontogeny of eyeblink conditioning. Studies of the development of physiological processes within the cerebellum have revealed striking ontogenetic changes in stimulus-elicited and learning-related neuronal activity. Neurons in the interpositus nucleus and Purkinje cells in the cortex exhibit developmental increases in neuronal discharges following the unconditioned stimulus (US) and in neuronal discharges that model the amplitude and time-course of the eyeblink CR. The developmental changes in CR-related neuronal activity in the cerebellum suggest that the ontogeny of eyeblink conditioning depends on the development of mechanisms that establish cerebellar plasticity. Learning and the induction of neural plasticity depend on the magnitude of the US input to the cerebellum. The role of developmental changes in the efficacy of the US pathway has been investigated by monitoring neuronal activity in the inferior olive and with stimulation techniques. The results of these experiments indicate that the development of the conditioned eyeblink response may depend on dynamic interactions between multiple developmental processes within the eyeblink neural circuitry. PMID- 11484995 TI - Parallel neural systems for classical conditioning: support from computational modeling. AB - Classical conditioning has been explained by two main types of theories that postulate different learning mechanisms. Rescorla and Wagner (1972) put forth a theory in which conditioning is based on the ability of the US to drive learning through error correction. Alternatively, Mackintosh (1973) put forth a theory in which the ability of the CS to be associated with the unconditioned stimulus is modulated. We have proposed a reconciliation of these two mechanisms as working in parallel within different neural systems: a cerebellar system for US modulation and a hippocampal system for CS modulation. We developed a computational model of cerebellar function in eyeblink conditioning based on the error correction mechanism of the Rescorla-Wagner rule in which learning-related activity from the cerebellum inhibits the inferior olive, which is the US input pathway to the cerebellum (Gluck et al., 1994). We developed a computational model of the hippocampal region that forms altered representations of conditioned stimuli based on their behavioral outcomes (Gluck & Myers, 1993; Myers et al., 1995). Overall, computational modeling and empirical findings support the idea that, at least in the case of eyeblink conditioning, there may be two different neural systems: the cerebellum which mediates US-based error correction and hippocampus which alters representations of CSs. PMID- 11484996 TI - An epistemologically arrogant community of contending scholars: a pre-Socratic perspective on the past, present, and future of the Pavlovian Society. AB - The paper begins with a statement of the Society's purpose and its pre-Socratic roots. The Society differs from other contemporary scientific and scientific professional societies in that it is thoroughly apolitical, unusually open to discussion and debate, and has had a restricted scholarly written impact. I then suggest and interpret six phases in the Society's history: (1) the pre-Socratic roots; (2) Pavlov and the young Gantt; (3) the Society's Gantt score of years; (4) the Joe McGuigan decade; (5) the Stewart Wolf era; (6) reforming the Society. I conclude with the hope that even if the content of the Society's interests changes, it will preserve the pre-Socratic approach against the various forms of intellectual barbarism that continue to arise. KEYWORDS: Pre-Socratics, disinterested discussion, conflict of ideas, contending scholars, Pavlovian procedures PMID- 11484997 TI - Discrimination reversal conditioning of an eyeblink response is impaired by NMDA receptor blockade. AB - In the present study we examined the effects of the specific NMDA receptor antagonist CPP on discrimination reversal learning in rabbits. We report two primary findings. First, the institution of NMDA receptor blockade had no effect on a learned discrimination. Second, after stimulus reversal, CPP treatment impaired acquisition of the discrimination reversal. This impairment manifested itself early in training as a retardation in acquisition of a CR to the new CS+ and late in training as an inability to suppress responsiveness to the new CS-. Given the comparability of the present results with previously published results for phenytoin-treated rabbits, we suggest that the effects of phenytoin on learning in this paradigm is at least in part mediated by its effects on NMDA receptors. We further suggest that these findings emphasize the need to better define the role of NMDA receptor activation and hippocampally-mediated circuits in a variety of associative learning paradigms. PMID- 11484998 TI - Is there a role for psychology in ulcer disease? AB - The discovery of the importance of bacterial factors in the etiology of ulcer disease has led to a neglect of psychological factors. However, both earlier theoretical and empirical approaches implicating these factors are supported by more recent studies, both epidemiological and experimental. While not ignoring the unquestioned role of Helicobacter, it is important for future research to recognize the multi-factorial nature of ulcer disease by which several factors, including stress, bacteria and non-steroid antiinflammatory drugs, may interact to drive a pre-pathology (erosions or ulcerations) to a pathological state (ulcer). Calls for general eradication programs should be cautioned in the light of possible unwanted side effects. PMID- 11484999 TI - Putative regulatory molecules in plants: evaluating melatonin. AB - Numerous classes of chemicals have been considered as regulators of various aspects of plant growth and development. In evaluating these putative regulatory molecules, plant biologists have encountered a number of challenges, including: the problem of quantifying substances present at trace levels in extremely complex mixtures; difficulty in obtaining and interpreting phenotypic responses to exogenous applications; and, until recently, the inability to selectively alter endogenous levels of these substances. Melatonin (N-acetyl 5 methoxytryptamine), a methoxylated indoleamine, is a potential regulatory molecule found in plants. Although no specific phenotype is currently associated with melatonin or its analogs in higher plants, it has important and unique biological activity in many other taxa, from algae to primates. In these organisms, melatonin functions as a night signal, coordinating responses to diurnal and photoperiodic environmental cues. We assess the process by which melatonin has been evaluated in plants so far and find that many of the methods for melatonin analysis, which have been adopted from animal studies, are inappropriate for use with plant materials. Thus, despite some interesting preliminary reports, research supporting the case for melatonin as a plant regulator is still in its infancy. PMID- 11485000 TI - Circadian urinary 6-sulphatoxymelatonin, cortisol excretion and locomotor activity in airline pilots during transmeridian flights. AB - Airline pilots divided into two groups of age (over and under 50 years) were studied before, during and after westbound (Madrid-Mexico City-Madrid, n = 12) and eastbound (Madrid-Tokyo-Madrid, n = 21) flights. A group of 10 age-matched people staying in Madrid were submitted to the same tests and served as a control group. Changes in urinary 6-sulphatoxymelatonin (6-aMTs) and free cortisol excretion (determined in 6-hr intervals) were measured by radioimmunoassay. Using wrist actigraphy, the circadian locomotor activity rhythm (LAR) was also monitored. Maximal baseline excretion of 6-aMTs occurred between 00:00 and 12:00 hr and maximal excretion of cortisol took place between 6:00 and 12:00 hr in the control group. Analysed globally, older pilots exhibited significantly lower values of 6-aMTs than younger ones. In both flight directions, pilots maintained the pattern of excretion of 6-aMTs, corresponding to baseline. The return flight to Madrid from Mexico and Tokyo coincided with a maximum in 6-aMTs excretion. Pilots kept the cortisol pattern found in the control group, with those over 50 years of age exhibiting significantly lower cortisol values than the younger ones. A 7-hr delay in acrophase of LAR after 2 days in Mexico City was found after cosinor analysis, and similar pre-flight values were found after returning to Madrid. An 8-9-hr acrophase advance of LAR was observed after arriving in Tokyo, with acrophase on the post-return flight day still being advanced 3 4 hr as compared to pre-flight values. Decreases in the amplitude of LAR in older pilots were found at Mexico City, as well as at Tokyo stopover and on post-flight day. Data confirm the occurrence of internal desynchronization in airline crewmembers after transmeridian flights. PMID- 11485001 TI - Protective effect of melatonin against adriamycin toxicity in the rat. AB - Adriamycin, an anthracyclinic antibiotic frequently used in quimioterapeutic treatments is highly toxic; it inhibits protein synthesis and provokes prooxidant effects. Melatonin has recently been shown to have high antioxidative properties. We tested if melatonin is able to neutralize the oxidative damage induced by a single dose (20 mg/kg, i.p.) of adriamycin preceded (3 days) and followed (7 days) by a low pharmacological dose (50 microg/kg, i.p.) of melatonin. After the administration of a single dose of adriamycin (20 mg/kg i.p.) to male Wistar rats, the reduced to oxidized glutathione (GSH/GSSG) ratio and the glutathione peroxidase (GPx, E.C. 1.11.1.9.) activity in the brain, intestine, heart, kidney, and lung were significantly reduced. When the treatment of adriamycin was preceded and followed by low pharmacological doses of melatonin, the decrease in the GSH/GSSG ratio was significantly reduced but the reduction in GPx activity was not attenuated. A significant increase in lipid peroxidation products was observed in brain, heart, and kidney tissues after a single administration of adriamycin, which was attenuated by pre- and post-treatment with a low pharmacological dose of melatonin. Our results demonstrate that oxidative damage induced by the antitumor drug, adriamycin, can be reduced by low pharmacological doses of melatonin. PMID- 11485002 TI - Physiological concentrations of melatonin and corticosterone affect phagocytosis and oxidative metabolism of ring dove heterophils. AB - A functional connection between the neuroendocrine and the immune systems has been established. Of particular interest is the finding that hormones such as melatonin and corticosterone are able to exert modulating effects on the immune function. Therefore, after determining the circadian rhythms of melatonin and corticosterone, we evaluated the in vitro effect of physiological concentrations of melatonin and corticosterone, separately and together, on the phagocytic function and superoxide anion levels of heterophils in ring dove (Streptopelia risoria). Trials were performed with concentrations corresponding to the nocturnal and diurnal levels reached by each of the hormones (50:300 pg/mL and 100:10 ng/mL for melatonin and corticosterone, diurnal:nocturnal, respectively). The phagocytes were incubated with the hormones both alone and concurrently. At the highest (nocturnal) concentration, melatonin augmented phagocytic function and at the same time inducing a fall in superoxide anion levels. At the highest (diurnal) concentration, corticosterone also enhanced phagocytic function, but without modifying the phagocyte oxidative metabolism. In the presence of both hormones, however, whether with nocturnal or diurnal concentrations, there was a greater increase in phagocytic function and a decrease in superoxide anion levels than was produced by either of the hormones alone. In conclusion, our findings suggest that melatonin and corticosterone may have an additive effect in the modulation of phagocytic function. PMID- 11485003 TI - Melatonin reduces UV-induced reactive oxygen species in a dose-dependent manner in IL-3-stimulated leukocytes. AB - Reactive oxygen species (ROS) are presumed to be involved in inflammatory UV reactions of the skin. This in vitro study was performed to investigate the suppressive effect of melatonin in interleukin-3 (IL-3) stimulated leukocytes. Neutrophilic granulocytes were isolated from EDTA-treated whole blood and placed in a phosphate-buffered saline (PBS) containing IL-3. Cell suspensions were either treated with PBS (control) or with increasing doses of melatonin (0.1, 0.5, 1, 2, 3, 5, 7.5, 10 mmol). One PBS solution was left unirradiated and the other nine solutions (PBS and melatonin) were irradiated with 750 mJ/cm2 UVB light (280-360 nm, max: 310 nm). Radical formation was measured by the chemiluminescence technique. UV-irradiated leukocytes showed a 5-fold higher radical formation than unirradiated leukocytes. Melatonin, in increasing doses in powers of ten, led to a maximum suppression of free radicals at 10 nmol (P= 0.01) and 1 mmol melatonin (P= 0.001), showing a biphasic, non-linear, dose response relationship. Melatonin, given in amounts of 0.1-10 mmol, led to a direct dose dependent suppression of ROS. Radical formation was suppressed significantly in a range from 0.5 to 10 mmol (P= 0.001). Melatonin is known to function as a radical scavenger and antioxidant; some of these melatonin effects may be receptor independent, while others may be receptor dependent. PMID- 11485004 TI - Acute modulation of somatostatin receptor function by melatonin in the rat frontoparietal cortex. AB - Since melatonin (N-acetyl-5-methoxytryptamine) decreases locomotor activity and rearing and increases grooming behavior in a similar manner as somatostatin (SRIF), we examined if melatonin could induce these changes through somatostatinergic neurotransmission in the rat frontoparietal cortex. Male Wistar rats (200-250 g) received a single injection of melatonin (25 microg/kg per day) subcutaneously (s.c.) and were sacrificed 5 hr later. Melatonin treatment increased the number of 125I-Tyr11-SRIF receptors in frontoparietal cortical membranes without any changes in the dissociation constant (Kd). The capacity of SRIF to inhibit basal and forskolin (FK)-stimulated adenylyl cyclase (AC) activity was increased in melatonin-treated rats as compared to the control animals. Melatonin administration also induced a lower AC activity, both under basal conditions and after stimulation of the enzyme via stimulatory guanine nucleotide-binding proteins (Gs), or directly with FK. Functional inhibitory guanine nucleotide-binding protein (Gi) activity was increased in frontoparietal cortical membranes from melatonin-treated rats when compared to controls. Western blot analyzes showed that melatonin administration did not alter the amount of the Gialpha1, or Gialpha3 subunits, but reduced Gialpha2 levels in frontoparietal cortical membranes. No significant changes in SRIF-like immunoreactivity content and SRIF mRNA levels were detected in this brain area after melatonin treatment. Administration of the melatonin receptor antagonist luzindole (10 mg/kg, s.c.) 30 min before melatonin injection did not change the melatonin-induced effects on the SRIF receptor effector system. In conclusion, the present results show that acute melatonin administration increases the activity of the SRIF receptor effector system and decreases Gialpha2 levels in the rat frontoparietal cortex. In addition, the coupling of Gs to AC is disturbed by melatonin. PMID- 11485005 TI - Seasonal rhythms of 6-sulphatoxymelatonin (aMT6s) excretion in female rats are abolished by growth of malignant tumors. AB - The effect of development and growth of malignant tumors on pineal melatonin production was studied in two different hormone-dependent tumor systems in female rats. Urinary excretion of 6-sulphatoxymelatonin (aMT6s), the metabolic end product of melatonin, which parallels its production, was determined by radioimmunoassay at fortnightly or monthly intervals over the period of 1 year in female F344 Fischer rats bearing chemically-induced mammary carcinomas and in BDII/Han rats with spontaneous endometrial carcinomas. Untreated Fischer rats and BDII/Han rats in which tumor growth was suppressed by treatment with a progestin served as controls. Based on the cosinor analysis, animals without tumors showed significant seasonal rhythms of aMT6s excretion, with peaks in August (Fischer rats) and in May (BDII/Han rats). In contrast, such rhythms were absent in animals with developing and manifest tumors. It is concluded that animals kept under constant environmental conditions still show seasonal rhythms of pineal activity. Tumor development and growth affect pineal activity leading to disturbance of these rhythms. PMID- 11485006 TI - Melatonin, vitamin E, and estrogen reduce damage induced by kainic acid in the hippocampus: potassium-stimulated GABA release. AB - Melatonin, vitamin E and estrogen have been shown to exert neuroprotective effects against kainic acid (KA)-induced damage in the hippocampus. The aim of the present study was to examine the changes in potassium-evoked gamma aminobutyric acid (GABA) release in the hippocampus of KA-treated rats and to test the possible protective effects of melatonin, vitamin E or estrogen. Following the treatment of mice with KA, a marked reduction in potassium-evoked [3H]GABA release was observed. Melatonin or estrogen prevented the reduction in potassium-evoked GABA release due to kainate administration. Vitamin E also exhibited some protective effect, but it was less than that provided by melatonin or estrogen. Melatonin, estrogen and, to a lesser extent, vitamin E reduce the physiological toxicity of KA. Since KA is believed to cause neuronal alterations via oxidative processes, it is assumed that the free radical scavenging and oxidative properties of melatonin, estrogen and vitamin E account for the protective effects of these agents. PMID- 11485007 TI - Melatonin rhythms in European sea bass plasma and eye: influence of seasonal photoperiod and water temperature. AB - The transduction of seasonal information from the environment (i.e., photoperiod and water temperature) into melatonin rhythms was studied in sea bass. Plasma and ocular melatonin (N-acetyl-5-methoxytryptamine) was determined in autumn, winter, spring and summer (experiment 1) under natural culture conditions, and in the summer and winter solstices under both natural and "6-month out-of-phase" photoperiods (experiment 2). At each sampling, 48 sea bass were sacrificed at a rate of 6 fish every 3 hr and the level of melatonin was determined in plasma and eye cup samples by ELISA. In experiment 1, significant diel changes were observed in plasma melatonin, with nocturnal melatonin varying from 144 pg/mL (summer) to 23 pg/mL (autumn), while diurnal melatonin remained low, around 8 pg/mL throughout the year. In experiment 2, the photoperiod length was shown to control the duration of the nocturnal melatonin rise, while the water temperature determined the amplitude of the melatonin rhythm. Ocular melatonin peaked during daytime in autumn and winter, but no significant changes were detected in summer and spring. In conclusion, plasma melatonin rhythms in sea bass reflect the pineal capacity to integrate seasonal information and supply precise calendar information, which may synchronize different physiological processes such as annual reproduction and feeding rhythms. PMID- 11485008 TI - Effect of melatonin on cellular energy depletion mediated by peroxynitrite and poly (ADP-ribose) synthetase activation in an acute model of inflammation. AB - DNA single-strand breakage and activation of the nuclear enzyme poly (ADP-ribose) synthetase (PARS) triggers an energy-consuming, inefficient repair cycle, which contributes to peroxynitrite-induced cellular injury. Recently, we proposed that during an acute model (pleurisy), cellular injury is mediated by peroxynitrite formation and consequent PARS activation. Here, we investigated whether in vivo melatonin treatment inhibits cellular injury induced by peroxynitrite production and PARS activation in macrophages collected from rats subjected to carrageenan induced pleurisy. Macrophages harvested from the pleural cavity exhibited a significant production of peroxynitrite, as measured by the oxidation of the fluorescent dye dihydrorhodamine 123. Furthermore, carrageenan-induced pleurisy caused a suppression of macrophage mitochondrial respiration, DNA strand breakage, activation of PARS, and reduction of cellular levels of NAD+. In vivo treatment with melatonin [12.5 or 25 or 50 mg/kg, intraperitoneally (i.p.), 30 min before carrageenan] significantly reduced peroxynitrite formation in a dose dependent manner and prevented the appearance of DNA damage, decrease in mitochondrial respiration, loss of cellular levels of NAD+, and PARS activation. Our study supports the view that the antioxidant and anti-inflammatory effect of melatonin is also correlated with the inhibition of peroxynitrite production and PARS activation. In conclusion, melatonin may be a novel pharmacological approach to prevent cell injury in acute inflammation. PMID- 11485009 TI - Melatonin in plant organs. AB - The indoleamine melatonin, a well-known animal chemical, has been identified in extracts from several plant species. The function of melatonin in plants is unknown. Two major functions of melatonin in animals are dark signaling and antioxidant protection. Fruit ripening was used as a model physiological process that involves changes in the oxidative status of an organ. Tomato fruits at various stages of ripeness were sampled. Morning glory (Pharbitis nil Choisy, cv. Violet) and tomato (Lycopersicon esculentum Mill., cv. T5 and Castlemart) organs were collected throughout a light/dark cycle to determine whether melatonin levels increased during the night. No consistent evidence was found that melatonin increased significantly in organs of these plants during the night, as it does in many animals. The melatonin content of the fruits generally increased during ripening up to the mature ripe stage and thereafter as the fruit became over ripe. PMID- 11485010 TI - Melatonin and 6-OHMS in high-intensity magnetic fields. AB - We determined if all-night exposure to 60 Hz magnetic fields at an intensity well within the upper range of occupational exposures (resultant intensity = 127.3 microTesla [muT]) resulted in suppression of melatonin or its major metabolite 6 hydroxymelatonin-sulfate (6-OHMS) in the first-void morning urine of 24 healthy male volunteers. Measures collected after continuous and intermittent magnetic field exposure test conditions were compared to similar measures collected after an equivalent no-exposure control condition. Urinary concentrations of melatonin and 6-OHMS did not differ as a function of type of magnetic field exposure, nor did they differ between field exposure and control conditions. These results demonstrate that the nocturnal secretion and metabolism of melatonin are not altered in humans by field exposure at an intensity over 600 times higher than that typically encountered in the home. PMID- 11485011 TI - Aging-dependent changes in the effect of daily melatonin supplementation on rat metabolic and behavioral responses. AB - Pineal melatonin secretion has been reported to commonly decrease with aging, whereas intra-abdominal adiposity, plasma insulin and plasma leptin levels tend to increase. We recently demonstrated that daily melatonin administration starting at middle age suppressed male rat intra-abdominal fat, plasma leptin and plasma insulin to youthful levels, suggesting that aging-related changes in pineal melatonin secretion and in energy regulation may be functionally related. Accordingly, we have now investigated the effects of daily melatonin treatment on energy regulation in young versus middle-aged male Sprague Dawley rats. Addition of melatonin to the drinking water (0.2 microg/mL) produced nocturnal and diurnal plasma melatonin concentrations in middle-aged rats (12 months) equivalent to those of young adult (5 months) rats. Administration of this melatonin dosage every day for 10 wk starting at 10 months of age suppressed (P < 0.01) relative intra-abdominal fat, non-fasted plasma insulin and plasma leptin by 27, 39, and 51%, respectively (vs. vehicle-treated controls). In contrast, administration of melatonin for 10 wk starting at 3 months of age did not significantly alter (P> 0.10) any of these parameters. The melatonin administration stimulated (102%, P < 0.001) behavioral responsiveness of the middle-aged rats in a test of response to novelty, restoring youthful levels, but did not significantly alter behavioral responsiveness of the young rats. These results suggest that suppression of intra abdominal adiposity and plasma leptin and insulin levels and stimulation of behavioral responsiveness in response to daily exogenous melatonin begins at middle age, coincident with and likely dependent upon the aging-associated decline in endogenous pineal melatonin secretion. These results further suggest that appropriate melatonin supplementation may potentially provide therapy or prophylaxis not only for the insulin resistance, increased intra-abdominal fat and resulting pathologies that occur with aging, but also for some aging associated behavioral changes. PMID- 11485012 TI - Melatonin: an antioxidant at physiological concentrations. PMID- 11485013 TI - Maturity onset diabetes of the young as a model for complex disease. PMID- 11485014 TI - JMM, past and present. PMID- 11485015 TI - G protein coupled receptor dimerization: implications in modulating receptor function. AB - Protein-protein interactions are involved in the regulation of a large number of biological processes. It is well established that a variety of cell surface receptors interact with each other to form dimers, and that this is essential for their activation. Although the existence of G protein coupled receptor dimers was predicted from early pharmacological and biochemical analysis, solid evidence supporting dimerization has come within the past few years following the cloning of G protein coupled receptor cDNAs. Using differential epitope tagging and selective immunoisolation of receptor complexes, dimerization of a number of G protein coupled receptors including members of the rhodopsin, secretin, and metabotropic glutamate receptor families have been reported. More recently fluorescence or bioluminescence resonance energy transfer techniques have been used to examine dimerization of these receptors in live cells. These studies have found that whereas in some cases there is an agonist induced increase in the level of dimers, in others there is a decrease or no change in dimer levels. Several recent studies have also reported the ability of related members of G protein coupled receptors to heterodimerize. These heterodimers exhibit distinct physical and functional properties. Examination of possible sites of interactions between receptors has implicated a role for extracellular, transmembrane and/or C terminal region in dimerization. The functional consequences of dimerization, explored mainly using mutant receptors, have demonstrated a role in modulating agonist affinity, efficacy, and/or trafficking properties. Thus dimerization appears to be a universal phenomenon that provides an additional mechanism for modulation of receptor function as well as cross-talk between G protein coupled receptors. PMID- 11485016 TI - Role of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in bone cell biology. AB - Receptor activator of nuclear factor (NF-kappaB) ligand (RANKL), its cellular receptor, receptor activator of NF-kappaB (RANK), and the decoy receptor osteoprotegerin (OPG) constitute a novel cytokine system. RANKL produced by osteoblastic lineage cells and activated T lymphocytes is the essential factor for osteoclast formation, fusion, activation, and survival, thus resulting in bone resorption and bone loss. RANKL activates its specific receptor, RANK located on osteoclasts and dendritic cells, and its signaling cascade involves stimulation of the c-jun, NF-kappaB, and serine/threonine kinase PKB/Akt pathways. The effects of RANKL are counteracted by OPG which acts as a soluble neutralizing receptor. RANKL and OPG are regulated by various hormones (glucocorticoids, vitamin D, estrogen), cytokines (tumor necrosis factor alpha, interleukins 1, 4, 6, 11, and 17), and various mesenchymal transcription factors (such as cbfa-1, peroxisome proliferator-activated receptor gamma, and Indian hedgehog). Transgenic and knock-out mice with excessive or defective production of RANKL, RANK, and OPG display the extremes of skeletal phenotypes, osteoporosis and osteopetrosis. Abnormalities of the RANKL/OPG system have been implicated in the pathogenesis of postmenopausal osteoporosis, rheumatoid arthritis, Paget's disease, periodontal disease, benign and malignant bone tumors, bone metastases, and hypercalcemia of malignancy, while administration of OPG has been demonstrated to prevent or mitigate these disorders in animal models. RANKL and OPG are also important regulators of vascular biology and calcification and of the development of a lactating mammary gland during pregnancy, indicating a crucial role for this system in extraskeletal calcium handling. The discovery and characterization of RANKL, RANK, and OPG and subsequent studies have changed the concepts of bone and calcium metabolism, have led to a detailed understanding of the pathogenesis of metabolic bone diseases, and may form the basis of innovative therapeutic strategies. PMID- 11485017 TI - Pathophysiology of limb girdle muscular dystrophy type 2A: hypothesis and new insights into the IkappaBalpha/NF-kappaB survival pathway in skeletal muscle. AB - Limb girdle muscular dystrophies (LGMDs) are a group of clinically heterogeneous genetic diseases characterized by progressive weakness and atrophy of scapular and pelvic muscles, with either a dominant or recessive autosomic mode of inheritance. The first symptoms of the disorder appear during the first 20 years of life and progresses gradually, and a walking disability develops 10-20 years later. The gene responsible for LGMD2A has been identified and encodes calpain 3, a protease expressed mainly in skeletal muscle. Apoptotic myonuclei were recently detected in muscular biopsy specimens of LGMD2A patients, and apoptosis was found to be correlated with altered subcellular distribution of inhibitory protein kappaBalpha (IkappaBalpha) and nuclear factor kappaB (NF-kappaB), resulting in sarcoplasmic sequestration of NF-kappaB. Calpain 3 dependent IkappaBalpha degradation was reconstituted in vitro, supporting a possible in vivo sequence of events leading from calpain 3 deficiency to IkappaBkappa accumulation, prevention of nuclear translocation of NF-kappaB, and ultimately apoptosis. Therefore calpain 3, present in healthy muscle as sarcoplasmic and nuclear forms, may control IkappaBalpha turnover and indirectly regulate NF-kappaB dependent expression of survival genes. Recent data reported from a new model of LGMD2A in mice and from other muscular disorders strengthen understanding of the molecular links between calpain 3 and the Ikappaalpha/NF-kappaB pathway. Finally, in light of the lack of apoptosis observed in inflammatory myopathies, a unifying model for the control of cell survival in muscle is proposed and discussed PMID- 11485018 TI - A genome-wide scan suggests a locus on chromosome 1q21-q23 contributes to normal variation in plasma cholesterol concentration. AB - To identify genes that influence plasma cholesterol, triglyceride, and high density and low-density lipoproteins concentrations we conducted a genome-wide scan using 354 polymorphic markers spaced at 10-cM intervals in 75 obese but otherwise normal human families. The results of the genome scan using sibling pair analysis of quantitative phenotypes suggested that 1q21-q23 contains a locus that influences plasma cholesterol concentration. Chromosome 12 gave evidence of linkage to plasma triglyceride concentration (D12SPAH) and chromosomes 3, 6, 7, 10, 11, 17, and 20 yielded additional evidence of linkage for lipid phenotypes at lower levels of statistical significance. Allele sharing for markers near prominent candidate genes was either very weakly related or unrelated to sibling similarity for lipid concentrations. Together these results suggest that genes with important roles in regulating normal cholesterol and triglyceride concentrations do not coincide with the location of previously known candidate genes. PMID- 11485019 TI - Single-strand conformation polymorphism analysis of the glucose transporter gene GLUT1 in maturity-onset diabetes of the young. AB - Maturity-onset diabetes of the young (MODY), an autosomal dominant, early-onset form of type-2 diabetes, is caused by mutations in five different genes all leading to defect(s) in the pancreatic beta cell. However, some patients with this form of diabetes do not bear a mutation in any of the known (MODY1-MODY5) loci, a notion prompting the search for new MODY genes. Clinical and genetic data point toward a defect in beta cell function in the majority of patients with MODY, and partners of the glucose-sensing device are reasonable functional candidates. The high-capacity glucose transporter GLUT2 has the ideal kinetic features for performing this task. However, complete GLUT2 deficiency in humans leads to hepato-renal glycogenosis (Fanconi-Bickel syndrome), and heterozygous GLUT2 mutations apparently behave in a recessive manner. Furthermore, in the human beta cell GLUT1 mRNA is predominant when compared to GLUT2 and glucose influx appears to be largely mediated by this low-Km transporter. Thus, we looked for the presence of sequence variants by polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) within the GLUT1 gene in 90 Italian pedigrees negative at the search for mutations in glucokinase (MODY2) and hepatocyte nuclear factor-1alpha (MODY3), the two genes responsible for about 60% of MODY cases in Italian children. We found three already described silent mutations and a new single base deletion in position -173 of the 5' regulatory region. The -173de1A variant, which was detected in the heterozygous or homozygous state in 30.8% of MODY patients examined and is located in a Nuclear Factor Y binding sequence, is not associated with hyperglycemia in affected relatives of MODY probands. In conclusion, it appears from these results that the glucose transporter gene GLUT1 is unlikely to play a major role in the etiology of MODY diabetes. PMID- 11485020 TI - Expression of human beta-defensin-1 promotes differentiation of keratinocytes. AB - Epithelial cells have been shown to express the antibiotic peptides human beta defensins-1 and 2. While beta-defensin-2 is known to be up-regulated by bacterial factors and proinflammatory mediators, the expression of beta-defensin-1 does not appear to be affected by these mediators. To determine the regulation and function of beta-defensin-1 we analyzed its expression upon stimulation of inflammatory mediators in vitro and ex vivo. In immortalized human cell lines (HaCaT) and nasal polyps beta-defensin-1 was not induced upon incubation with bacteria or proinflammatory mediators, suggesting that the inertness of beta defensin-1 expression levels is not the result of the shortcoming of HaCaT cells. As proliferation and regeneration play an important role at sites of inflammation, we examined the expression level of beta-defensin-1 in relation to the differentiation and proliferation of HaCaT cells. beta-defensin-1 mRNA levels remained low during proliferation but were highly induced upon differentiation. In contrast, beta-defensin-2 expression was unaffected under these conditions. To examine the function of beta-defensin-1 in cellular proliferation and differentiation processes beta-defensin-1 was overexpressed in keratinocytes. Protein expression analysis of the differentiation marker keratin 10 revealed that its expression is highly induced in the presence of increased concentrations of beta-defensin-1. Hence our data indicate that high expression of beta-defensin 1 promotes cell differentiation processes of keratinocytes. PMID- 11485021 TI - p53 mutations are rare events in recurrent cervical cancer. AB - Mutations of the p53 gene have been shown to be associated with aggressive growth behavior and increased recurrence rates for certain tumors. Primary cervical cancers contain oncogenic human papillomaviruses (HPV) in more than 90% of cases and usually possess wild-type p53 alleles. Cervical cancer cells contain detectable levels of functional p53 protein despite of the expression of the HPV E6 protein, which can induce p53 degradation. Thus, inactivation of p53 by somatic mutation should have functional consequences in HPV-positive cancers. We investigated whether p53 mutations play a role in the recurrence of the disease by analyzing p53 status in 18 biopsy specimens from recurrent cervical cancers. Only one of these (5.6%) contained a p53 mutation, as assessed by a sensitive yeast functional assay that detects mutations of the p53 mRNA between codons 52 and 364. These results indicate that p53 mutations are rare events in recurrent cervical carcinomas, and that somatic mutations of p53 do not provide cervical cancer cells with a selective growth advantage for recurrence. PMID- 11485022 TI - The plasminogen activator inhibitor 1 4G/5G polymorphism is not associated with longevity: a study in octogenarians. AB - Accumulating evidence suggests that plasma levels of the plasminogen activator inhibitor 1 (PAI-1) may modulate the risk of coronary artery disease. The regulation of PAI-1 levels underlies not only environmental but also genetic influences. The 4G/5G polymorphism of the PAI-1 gene has recently gained additional relevance as a possible cardiovascular risk factor, as the 4G allele may be associated with enhanced expression of the PAI-1 gene. This retrospective cohort study examined the effect of the PAI-1 4G/5G genotype on longevity among 205 subjects aged 80 years and older. Such studies in larger cohorts have recently become available along with new methods for the rapid and easy determination of gene polymorphisms. We utilized a light-cycler assisted method which is a fast and flexible method of analyzing the PAI-1 4G/5G polymorphism on the gene level. In these 205 persons the 4G/5G allele was found in 96 persons (47%), the 4G/4G variant in 62 (30%), and the 5G/5G allele in 47 (23%). These data are similar to the allele distribution described in other large cohorts not restricted to old age. Thus the results of this study are not suggestive of an important contribution of the PAI-1 genotype on total mortality. PMID- 11485023 TI - Polymorphisms in human apolipoprotein(a) kringle IV-10 and coronary artery disease: relationship to allele size, plasma lipoprotein(a) concentration, and lysine binding site activity. AB - Elevated plasma levels of lipoprotein(a) [Lp(a)] represent a major independent risk factor for the development of atherosclerosis. The kringle IV type 10 of apolipoprotein(a) [apo(a)] is the primary lysine binding site (LBS) of Lp(a) and is associated with lesion formation in transgenic mice. The purpose of this study was to search for mutations in the apo(a) kringle IV type 10 which could alter the LBS activity of Lp(a) from patients with coronary artery disease. We found the DNA region of kringle IV type 10 of apo(a) to be mutable but relatively well preserved in the Spanish population. We identified a novel mutation which probably leads to a truncated form of apo(a) in a patient heterozygous for the mutation and with low lysine binding activity and low plasma Lp(a) concentration. Two other mutations have been previously identified in humans, the substitutions W81R and M75T. The W81R was not found in our sample, but the M75T mutation was present in 43% of patients with coronary artery disease and 23% of age-matched controls. The genotype TT conferred a significant risk for myocardial infarction (odds ratio 2.53). This association was not due to linkage disequilibrium with kringle IV repeats. The M75T polymorphism was not associated with the LBS function of apo(a), but it influenced plasma Lp(a) concentration. PMID- 11485024 TI - Interleukin-6 gene polymorphisms and susceptibility to myocardial infarction: the ECTIM study. Etude Cas-Temoin de l'Infarctus du Myocarde. AB - There is growing evidence that interleukin (IL) 6 plays an important role in the atherosclerotic process because of its role in mediating immune and inflammatory responses and inducing cell proliferation. The present study examined whether molecular variations at the IL-6 locus are involved in the predisposition to myocardial infarction. The entire coding region, 1,158 bp of the 5' flanking region and 237 bp of the 3' flanking region of the IL-6 gene were screened. We detected three nucleotide substitutions in the 5' region at positions -174 (G/C), -572 (G/C), and -596 (G/A) from the transcription start site, and one insertion/deletion in the 3' region at position +528 after the Stop codon. These polymorphisms were genotyped in the Etude Cas-Temoin de l'Infarctus du Myocarde study comparing male patients (n=640) and age-matched controls (n=719) from Northern Ireland and France. The IL-6/G-174C and IL-6/G-596A polymorphisms were in nearly complete association. Carriers of the IL-6/-174 C allele were more frequent in patients than in controls. The population-adjusted odds ratio for myocardial infarction associated with genotype CC+CG vs. GG was estimated as 1.34. In French patients the number of coronary arteries with greater than 50% stenosis was assessed by angiography. The IL-6/-174 C allele was more frequent in patients with two or fewer stenosed vessels than in patients with three-vessel lesions. These results suggest that genetic variation at the IL-6 locus is associated with susceptibility to myocardial infarction, especially events occurring on less extended lesions. These findings would be compatible with a lower IL-6 secretion associated with the IL-6/-174 C allele, itself or in combination with other promoter polymorphisms, leading to more unstable plaques. PMID- 11485025 TI - Shock waves: a novel method for cytoplasmic delivery of antisense oligonucleotides. AB - Intracytoplasmic delivery of oligonucleotides (ODN) can improve ODN-based strategies such as the antisense approach and the use of immunostimulatory CpG dinucleotide containing ODN. Shock waves are established for the treatment of nephrolithiasis and other diseases. Here we describe the use of shock waves as a new physical method for the direct transport of antisense ODN into the cytoplasm and the nucleus of cells. Human peripheral blood mononuclear cells together with antisense ODN were exposed to shock waves generated by an electrohydraulic lithotripter. ODN uptake was examined by flow cytometry and fluorescence microscopy. By optimization of physical parameters we achieved the transfer of high amounts of ODN which were detected within less than 5 min after shock wave exposure, with viability of cells higher than 95%. Transfection of human peripheral blood mononuclear cells with an antisense ODN directed against tumor necrosis factor (TNF) alpha resulted in a reduction in lipopolysaccharide-induced TNF production by 62% (n=5, P=0.006). Specificity of TNF suppression was confirmed with a four-mismatch oligonucleotide. Positive atmospheric pressure abolished antisense-mediated inhibition of TNF synthesis by blocking shock wave induced cavitation and formation of oscillating air bubbles. Electroporation was less effective. The use of shock waves is thus an efficient physical tool for ODN delivery to cells. Shock waves may allow the evaluation of target proteins in cell types difficult to transfect with other methods and thus may improve the antisense technique for the analysis of unknown genes. PMID- 11485026 TI - Expression pattern of AP-2 transcription factors in cervical cancer cells and analysis of their influence on human papillomavirus oncogene transcription. AB - The AP-2 family of transcription factors consists of three known members, namely AP-2alpha, AP-2beta, and AP-2gamma. In experimental systems AP-2 factors possess tumor suppressor-like activities, and alterations in the AP-2 expression pattern have been described for some tumor entities. In addition, AP-2 has been implicated in the transcriptional control of human papillomaviruses (HPVs). We investigated here the expression pattern of AP-2alpha, AP-2beta, and AP-2gamma, as well as that of the cellular AP-2 target gene c-erbB-2, in a series of cervical cancer cell lines. In addition, we analyzed the influence of AP-2 factors on the activity of the HPV16 and HPV18 E6/E7 oncogene promoter. We found that, with the exception of HPV-negative C33A cells, all investigated cervical cancer cell lines expressed all three AP-2 family members, although at varying levels. No linear correlation between AP-2 and c-erbB-2 levels was observed. Although AP-2alpha, AP-2beta, and AP-2gamma can activate the c-erbB-2 promoter in reporter gene assays, they do not stimulate the HPV16 or HPV18 E6/E7 promoter. These results indicate that, although a rare event, loss of AP-2 expression occurs in cervical cancer cells. Moreover, AP-2alpha, AP-2beta, and AP-2gamma are neither sufficient nor required to activate the viral E6/E7 promoter. PMID- 11485027 TI - The Tet-On system in transgenic mice: inhibition of the mouse pdx-1 gene activity by antisense RNA expression in pancreatic beta-cells. AB - To elucidate the function of pancreas duodenal homeobox 1 (PDX-1; insulin promoter factor 1/somatostatin transcription factor 1/islet duodenum homeobox 1/insulin upstream factor 1) in differentiated beta-cells of adult animals we generated transgenic mice using the Tet-On system. Inducible expression of an antisense RNA should down-regulate the PDX-1 protein. The selective and continuous inhibition of PDX-1 gene expression should impair the expression of PDX-1 dependent beta-cell specific genes. A gene switch such as the Tet-On system provides a powerful tool to analyze eukaryotic gene expression and function in transgenic mice. The original Tet system contained two transcriptional units, transactivator and target of control, on two plasmids. We combined the two transcriptional units on a single DNA molecule. The transactivator was placed under control of the mouse insulin promoter. The tet responsive element, driving the gene of interest, was inserted further down-stream into the same vector. The tet regulatory system in this approach permitted a tissue-specific and a doxycycline-inducible control of PDX-1 expression in transgenic mice. The expression of glucose transporter 2 and glucokinase was markedly reduced in dox treated transgenic mice. In contrast, the number of insulin- and amylin expressing cells was only slightly decreased, whereas the expression of glucagon was increased distinctly in islets of these mice. Furthermore, the exposure to doxycycline resulted in a progressive impairment of glucose tolerance. The characterization of our transgenic mouse model demonstrates the suitability of the Tet-On system for analyzing physiological consequences emerging from a stepwise decrease in a given protein. Using this system we confirmed the essential role of PDX-1 in pancreatic islets and demonstrated that an antisense mediated PDX-1 deficiency provokes a beta-cell dysfunction. PMID- 11485028 TI - Use of degenerate oligonucleotide primed PCR (DOP-PCR) for the genotyping of low concentration DNA samples. AB - Degenerate oligonucleotide primed amplification (DOP-PCR) is an efficient method for performing whole genome amplification. We analysed the yield of DNA using this technique starting with various quantities of material. We used DOP-PCR products to genotype single nucleotide polymorphisms and insertion/deletion polymorphisms. DOP-PCR also proved usable for SSCP analysis. PMID- 11485029 TI - Submerged gel electrophoresis on Spreadex gels--a new method for APC gene mutation detection. AB - Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease. Patients with FAP develop hundreds to thousands of adenomatous polyps in the colon and rectum during their 2nd or 3rd decades, and one or more of them progress to cancer if left without surgical treatment. The gene responsible for FAP was identified in 1991 and termed the APC (adenomatous polyposis coli) gene. Following identification of APC, a number of germ-line mutations responsible for the development of the disease were found. The purpose of this study was to determine the usefulness of a new method, submerged gel electrophoresis, in the detection of the most-frequent mutation of the APC gene [5-base pair (bp) deletion in codon 1309], especially in the presymptomatic diagnosis of FAP. Genomic DNAs were isolated from peripheral blood of patients and their relatives. We used two methods, electrophoresis on polyacrylamide gel and submerged gel electrophoresis, for the identification of APC gene codon 1309 mutation. After only 110 min PCR fragments of 91 bp and 86 bp (5-bp deletion) were completely resolved on a Spreadex EL300 gel. Our results showed that electrophoresis using Spreadex gels provides a simple and rapid non-radioactive method for determination of the most-frequent germ-line mutations in the APC gene. PMID- 11485030 TI - Absence of mutations in human ubiquitin fusion-degradation protein gene in tetralogy of Fallot. AB - Congenital defects in human chromosome 22q11 deletion syndromes are associated with the 3rd and 4th pharyngeal pouch during fetal development. In the cardiovascular system, these disorders are usually apparent as conotruncal heart defects and aortic arch anomalies. UFD1L, a gene that is downregulated in dHAND deficient mice, expressed in the mouse embryo at the branchial arch and mapped to human chromosome 22q11, has recently been strongly suspected to be responsible for the phenotypes expressed in 22q11 deletion syndromes. Its putative causal role in relevant congenital cardiovascular malformations was studied by gene dosage analysis, mutation screening and sequence analyses. Sixty cases of tetralogy of Fallot with no detectable chromosome deletion at 22q11 or 10p13 were examined, including 51 cases of simple tetralogy of Fallot, and 9 cases of tetralogy of Fallot with pulmonary atresia. None of these patients revealed deletion limited to a portion of the UFD1L gene. Although mobility shift was found by heteroduplex analysis in 24 cases at exon 4 and flanking sequences, further sequence analysis demonstrated only two silent nucleotide variations and a single nucleotide polymorphism in intron 4. Our data suggest that, although the UFD1L gene is mapped to 22q11 and is expressed during early murine development at both cardiac and cranial neural crests, it is not responsible for the majority of tetralogy of Fallot cases in humans. PMID- 11485031 TI - Priming of immune responses to hepatitis B surface antigen with minimal DNA expression constructs modified with a nuclear localization signal peptide. AB - Nuclear localization signal (NLS) peptides conjugated to DNA increase transfection efficiency in vitro. We tested in mice whether conjugation of NLS peptides to DNA vaccines enhances their immunogenicity after intramuscular injection or gene gun mediated intradermal delivery. We constructed the plasmid pMOK-HBsAY that contains a transcription unit encoding hepatitis B surface antigen (HBsAg) and bacterial sequences for amplification of plasmid DNA. From this plasmid we derived the minimal expression construct pMOK-HBsAY-MIDGE, a covalently closed linear DNA that contains only the HBsAg transcription unit. Both constructs stimulated similar (predominantly IgG1) antibody response to HBsAg after gene gun immunization. In contrast, pMOK-HBsAY plasmid DNA was more efficient than pMOK-HBsAY-MIDGE DNA in priming predominantly IgG2a antibody responses to HBsAg after intramuscular injection. Both constructs efficiently primed cytotoxic T lymphocyte responses after intramuscular immunization. When a NLS peptide was coupled to the pMOK-HBsAY-MIDGE DNA, HBsAg transfection efficiency in vitro and priming of antibody responses to HBsAg after intramuscular (but not gene gun mediated) injection was enhanced 10- to 15-fold. These data show: (a) MIDGE constructs can be used as DNA vaccines indicating that bacterial sequences are not essential cofactors; and (b) in intramuscular (but not gene gun mediated) delivery the immunogenicity of a MIDGE-based vaccine is enhanced by coupling NLS peptides to the vector DNA. PMID- 11485032 TI - Modulation of GABAA receptor-mediated currents by phenazepam and its metabolites. AB - The effects of 7-bromo-5-(2-chlorophenyl)-1,3-dihydro-2H-1,4-benzodiazepin-2-one (phenazepam, PNZ), a 1,4-benzodiazepine derivative, and its physiological metabolites on GABA-activated whole-cell currents were studied in enzymatically isolated rat Purkinje neurones. PNZ, its hydroxylated metabolite (HPNZ) and a reference benzodiazepine, diazepam, potently enhanced (up to 200% of control) peak amplitude of currents activated by 10 microM GABA with EC50s of 6.1 +/- 0.8, 10.3 +/- 1.4 and 13.5 +/- 1.9 nM respectively. Both PNZ and HPNZ caused a parallel leftwards shift of the concentration/effect relationship for GABA. Another metabolite, 6-bromo-(2-chlorophenyl) quinazoline-2-one (QNZ), augmented responses to 10 microM GABA with a maximal efficacy similar to that of the 1,4benzodiazepines tested, although its EC50 was 2.4 +/- 0.2 microM. A further metabolite, 5-bromo-(2-chlorophenyl)-2-aminobenzophenone (ABPH), had only minimal effects on the responses elicited by 10 microM GABA. Incubation with QNZ and ABPH had biphasic effects on the concentration/effect relationship for GABA. These compounds enhanced peak amplitudes of currents activated by low concentrations of GABA, but inhibited responses to saturating concentrations of the agonist. This effect could, in part, be explained by the acceleration of the desensitisation process by those substances. It is concluded that both PNZ and HPNZ can be referred to as full positive modulators of GABAA receptors and that they are primarily responsible for GABAergic effects of therapeutic doses of PNZ. PMID- 11485033 TI - Novel histaprodifen analogues as potent histamine H1-receptor agonists in the pithed and in the anaesthetized rat. AB - We have shown previously that histaprodifen and its Nalpha-substituted analogues methylhistaprodifen and dimethylhistaprodifen are highly potent H1-receptor agonists in vivo. The aim of the present study was to examine the influence of four newly synthesized histaprodifen analogues, 3-fluoro-methylhistaprodifen (1), Nalpha-imidazolylethylhistaprodifen (2), bis-histaprodifen (3) and Nalpha-methyl bis-histaprodifen (4), on the cardiovascular system in the pithed and in the anaesthetized rat. In pithed and vagotomized rats, diastolic blood pressure (which was increased to 80-85 mmHg by vasopressin infusion) was decreased dose dependently by methylhistaprodifen (the reference compound) and by compounds 1-4. The maximum decrease was about 47-50 mmHg for methylhistaprodifen and compounds 1, 2 and 3. Their potencies, expressed as pED50 (the negative logarithm of the dose in mole per kilogram body weight that decreased diastolic blood pressure by 25 mmHg), were 8.31, 8.23, 8.26 and 7.84, respectively. With compound 4 the maximal effect was not achieved at doses up to 1 micromol/kg (the latter dose decreased blood pressure by about 30 mmHg; pED50 approximately 6.5). The vasodepressor effect of the five compounds was attenuated by the H1-receptor antagonist dimetindene (1 micromol/kg) but was not changed by combined administration of the H2- and H3-receptor antagonists ranitidine and thioperamide (1 micromol/kg each), by combined administration of the alpha1- and alpha2 adrenoceptor antagonists prazosin and rauwolscine (1 micromol/kg each) or by the beta-adrenoceptor antagonist propranolol (3 micromol/kg). In anaesthetized rats methylhistaprodifen and compounds 1-4 induced almost the same fall in blood pressure as in pithed and vagotomized animals; the effects were sensitive to blockade by dimetindene (1 micromol/kg). Higher doses of compounds 1 and 2 (1 micromol/kg) increased heart rate in pithed and vagotomized rats in a manner sensitive to propranolol (3 micromol/kg) but insensitive to dimetindene (3 micromol/kg). The same dose of methylhistaprodifen and of compounds 3 and 4 failed to affect heart rate. We conclude that the agonistic potency of compounds 1 and 2 at H1-receptors in the cardiovascular system of the rat equals that of methylhistaprodifen, the most potent histamine H1-receptor agonist available until recently. Compounds 1 and 2 exhibit sympathomimetic activity at high doses. PMID- 11485034 TI - Characterization of antidepressant-like effects of p-synephrine stereoisomers. AB - We previously reported that p-synephrine has antidepressant-like activity in the murine models of forced swimming and tail suspension. In the present study, we characterized antidepressant-like effects of p-synephrine stereoisomers in both in vivo and in vitro systems. In the tail suspension test, S-(+)-p-synephrine (3 mg/kg, p.o.) reduced the duration of immobility, while R-(-)-p-synephrine (0.3-3 mg/kg, p.o.) had no effect. S-(+)-p-synephrine (0.3, 1 and 3 mg/kg, p.o.) and R-( )-p-synephrine (1 mg/ kg and 3 mg/kg, p.o.) significantly reversed the reserpine (2.5 mg/kg, i.p.)-induced hypothermia. S-(+)-p-synephrine was more effective than R-(-)-p-synephrine in inhibition of both [3H]noradrenaline uptake in rat cerebral cortical slices (maximal inhibition 85.7 +/- 7.8% vs. 59.8 +/- 4.3%; EC50 5.8 +/- 0.7 microM vs. 13.5 +/- 1.2 microM) and [3H]nisoxetine binding (Ki 4.5 +/- 0.5 microM vs. 8.2 +/- 0.7 microM). In contrast, R-(-)-p-synephrine was more effective than S-(+)-p-synephrine in stimulation of [3H]noradrenaline release from rat cerebral cortical slices (maximal stimulation 23.9 +/- 1.8% vs. 20.1 +/- 1.7%; EC50 8.2 +/- 0.6 microM vs. EC50 12.3 +/- 0.9 microM). The stimulatory effect of R-(-)-p-synephrine on [3H]noradrenaline release was inhibited by nisoxetine (100 nM), but tetrodotoxin (1 microM) and elimination of extracellular calcium had no effect. It is suggested that S-(+)-p-synephrine has more effective antidepressant-like activity than R-(-)-p-synephrine. PMID- 11485035 TI - Pharmacological profile of the mechanisms involved in the external carotid vascular effects of the antimigraine agent isometheptene in anaesthetised dogs. AB - The present study set out to investigate the external carotid vascular effects of isometheptene in vagosympathectomised dogs, anaesthetised with pentobarbital. One minute intracarotid (intra-arterial; i.a.) infusions of isometheptene (10, 30, 100 and 300 microg/min) produced dose-dependent decreases in external carotid blood flow, without affecting blood pressure or heart rate. The vasoconstrictor responses to 100 microg/min and 300 microg/min of isometheptene were clearly attenuated in animals pretreated with reserpine (5,000 microg/kg). Moreover, after prazosin (an alpha1-adrenoceptor antagonist; 100 microg/kg), the responses to isometheptene remained unaltered in untreated as well as reserpine-pretreated dogs. In contrast, the responses to isometheptene were attenuated by rauwolscine (an alpha2-adrenoceptor antagonist; 300 microg/kg) in untreated animals, and were practically abolished in reserpine-pretreated dogs. Further investigation into the specific alpha2-adrenoceptor subtypes, using selective antagonists, showed that BRL44408 (alpha2A) and MK912 (alpha2C) markedly attenuated this response, while imiloxan (alpha2B) was ineffective. The involvement of 5-HT1B and 5-HT1D receptors seems highly unlikely since antagonists at 5-HT1B (SB224289) and 5-HT1D (BRL15572) receptors (both at 300 microg/kg) were ineffective. On this basis, it is concluded that isometheptene-induced canine external carotid vasoconstriction is mediated by both indirect (a tyramine-like action) and direct (acting at receptors) mechanisms, which mainly involve alphaA- and alpha2C-adrenoceptors, while the involvement of alpha1-adrenoceptors seems rather limited. PMID- 11485036 TI - Effects of the cardioselective KATP channel blocker HMR 1098 on cardiac function in isolated perfused working rat hearts and in anesthetized rats during ischemia and reperfusion. AB - It has been argued that activation of KATP channels in the sarcolemmal membrane of heart muscle cells during ischemia provides an endogenous cardioprotective mechanism. In order to test whether the novel cardioselective KATP channel blocker HMR 1098 affects cardiac function during ischemia, experiments were performed in rat hearts during ischemia and reperfusion. Isolated perfused working rat hearts were subjected to 30 min of low-flow ischemia in which the coronary flow was reduced to 10% of its control value, followed by 30-min reperfusion. In the first set of experiments the hearts were electrically paced at 5 Hz throughout the entire protocol. At the end of the 30-min ischemic period the aortic flow had fallen to 44 +/- 2% (n=8) of its nonischemic value in vehicle treated hearts, whereas in the presence of 0.3 micromol/l and 3 micromol/l HMR 1098 it had fallen to 29 +/- 7% (n=5, not significant) and 8 +/- 2% (n=12, P<0.05), respectively. Glibenclamide (3 micromol/l) reduced the aortic flow to 9.5 +/- 7% (n=4, P<0.05). In control hearts the QT interval in the electrocardiogram shortened from 63 +/- 6 ms to 36 +/- 4 ms (n=10, P<0.05) within 4-6 min of low-flow ischemia. This shortening was completely prevented by 3 micromol/l HMR 1098 (60 +/- 5 ms before ischemia, 67 +/- 6 ms during ischemia, n=9, not significant). When rat hearts were not paced, the heart rate fell spontaneously during ischemia, and HMR 1,098 (3 micromol/l) caused only a slight, statistically non-significant reduction in aortic flow during the ischemic period. In order to investigate whether HMR 1098 shows cardiodepressant effects in a more pathophysiological model, the left descending coronary artery was occluded for 30 min followed by reperfusion for 60 min in anesthetized rats. Treatment with HMR 1098 (10 mg/kg i.v.) had no statistically significant effects on mean arterial blood pressure and heart rate during the control, ischemia and reperfusion periods. At the end of the reperfusion period, aortic blood flow was slightly reduced by HMR 1098, without reaching statistical significance (two-way analysis of ANOVA, P=0.15). Myocardial infarct size as a percentage of area at risk was not affected by HMR 1098 (vehicle: 75 +/- 3%, HMR 1098: 72 +/- 2%, n=7 in each group). In conclusion, cardiodepressant effects of HMR 1098 were observed only in isolated perfused working rat hearts which were continuously paced during global low-flow ischemia. In the model of anesthetized rats subjected to regional ischemia, HMR 1098 had no significant effect on cardiac function or infarct size. PMID- 11485037 TI - Failure of endothelin-1 to activate store-operated Ca2+ channels by lack of mobilization from intracellular Ca2+ stores in cultured bovine adrenal chromaffin cells. AB - We have recently shown that in addition to L-type voltage-operated Ca2+ channel (VOC), endothelin-1 (ET-1) stimulation opens two types of Ca2+-permeable nonselective cation channels [designated nonselective cation channel-1 (NSCC-1) and NSCC-2]. However, in this Ca2+ entry, the involvement of store-operated Ca2+ channel (SOCC), which is suggested to exist in chromaffin cells, was unclear. Those NSCCs as well as SOCC can be pharmacologically discriminated using Ca2+ channel blockers such as SK&F 96365 and LOE 908. To clarify whether SOCC should actually exist and play a role in Ca2+ entry in chromaffin cells stimulated with ET-1, we examined the effects of removal of extracellular Ca2+, thapsigargin (TG, an inhibitor of endoplasmic reticulum Ca2+-ATPase), LOE 908 and SK&F 96365 on cytosolic free Ca2+ concentrations ([Ca2+]i) in cultured bovine adrenal chromaffin cells. After the cells were exposed to Ca2+-free medium followed by exposure to TG to deplete Ca2+ from the intracellular Ca2+ store, restoration of extracellular Ca2+ caused a gradual increase in [Ca2+]i (to about 200% of control). The increase was unaffected by LOE 908, but completely abolished by SK&F 96365. In the Ca2+-free medium, no increase in [Ca2+]i by ET-1 was observed, but the subsequent restoration of extracellular Ca2+ induced a rapid increase in [Ca2+]i (to the same level of [Ca2+]i as that evoked by ET-1 in the normal medium (1.0 mM Ca2+)). Since SK&F 96365 is also a blocker of SOCC, these results indicate that in bovine adrenal chromaffin cells, Ca2+ entry through SOCC (Ca2+ influx through the capacitative Ca2+ entry system) occurs but is comparably weak, and that it virtually does not work on the stimulation of ET-1. PMID- 11485038 TI - Glucose-induced insulin secretion is potentiated by a new imidazoline compound. AB - Sulfonylureas stimulate insulin secretion independent of the blood glucose concentration. This can lead to hypoglycaemia in type 2 diabetic patients. Over the last years a number of imidazoline derivatives have been identified that stimulate insulin secretion in a more glucose-dependent way. In agreement with this, our aim was to generate imidazoline derivatives with a potential for the treatment of type 2 diabetic patients. We developed the compound 2-[4-(4 chlorophenyl)-3-(2-methoxyethoxy)-2-naphthalenyl]-4,5-dihydro-1-H-imidazole monohydrochloride (LY389382) with an imidazoline moiety and investigated its effects on glucose-dependent insulin secretion in a beta-cell line, isolated rat islets and in vivo. We could demonstrate that LY389382 induces insulin secretion in MIN6 cells and rat islets in a glucose-dependent manner (EC50=1.1 microM and 0.3 microM, respectively). Furthermore during hyperglycaemia LY389382 increased insulin secretion in a dose-dependent manner in healthy rats, whereas the compound had no effect at euglycemia in a tenfold higher dosage. After 7 days of treatment of Zucker Diabetic Fatty [ZDF/ (Gmi/fa)] rats with LY389382 with a dose of 15 mg/kg twice daily the blood glucose concentration was reduced from 22.7 +/- 1.7 mM to 16.6 +/- 2.3 mM. During the same time period the glucose concentration increased from 21.7+/-1.7 mM to 28.9 +/- 1.3 mM in the vehicle-treated group (P<0.05). The drop of the insulin level was also inhibited by LY389382 in ZDF rats. In contrast to other well-characterised imidazolines that have been shown to induce a glucose-dependent insulin secretion only within a limited range of concentrations, LY389382 stimulates insulin secretion over a concentration range of at least two log units in a glucose-dependent manner. These data suggest that this imidazoline compound has a potential for the treatment of type 2 diabetes. PMID- 11485039 TI - Bradykinin causes inhibition of methacholine-induced bronchoconstriction in vivo in mice. AB - In this study the influence of bradykinin on airway responses was investigated in anaesthetised and ventilated mice. Airway resistance in mice was monitored using whole body plethysmography. Intravenous (i.v.) administration of bradykinin (4-40 microg/kg) did not cause a direct effect on airway resistance. Also pretreatment with propranolol (1 mg/kg, i.v.), atropine (1 mg/kg, i.v.) or indomethacin (5 mg/kg, i.v.) did not result in any effect of intravenous bradykinin on baseline airway resistance. However, i.v. bradykinin (4-40 microg/kg) caused a dose dependent inhibition of the (0.5 mg/kg, i.v.) methacholine-induced bronchoconstriction, with an ED50 value of 3.4 +/- 0.4 microg/kg. The maximal inhibition of the bronchoconstrictor response to methacholine was 65.5 +/- 2.0%. The inhibition of the methacholine-induced bronchoconstriction by bradykinin could be prevented by treatment with the B2 receptor antagonist icatibant (Hoe 140, 0.13 mg/kg, i.v.). Also pretreatment with either propranolol (1 mg/kg, i.v.), L-NAME (30 mg/kg, i.v.) or indomethacin (5 mg/kg, i.v.) completely blocked the inhibition of the methacholine-induced bronchoconstriction by bradykinin. The inhibition of the methacholine-induced bronchoconstriction after bradykinin was not affected by the NK1 receptor antagonist RP 67580 (17.5 microg/kg, i.v.). In conclusion, the results of this study demonstrate that bradykinin causes a dose dependent inhibition of the methacholine-induced bronchoconstriction in vivo in mice. This response is B2 receptor-mediated and at least involves the activation of beta-adrenoceptors and the synthesis of nitric oxide and cyclo-oxygenase products. PMID- 11485040 TI - Nitric oxide induces hydroxyl radical generation in rat hearts via depolarization induced nitric oxide synthase activation. AB - We examined the effect of NG-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, on extracellular potassium ion concentration ([K+]o) and induced hydroxyl free radical (.OH) generation by an in vivo microdialysis technique. A flexibly mounted microdialysis technique was used to detect the generation of .OH in in-vivo rat hearts. The microdialysis probe was implanted in the left ventricular myocardium of anesthetized rats and tissue was perfused with Ringer's solution through the microdialysis probe at a rate of 1.0 microl/min. To measure the level of .OH, sodium salicylate in Ringer's solution (0.5 nmol/microl per min) was infused directly through a microdialysis probe to detect the generation of .OH as reflected by the nonenzymatic formation of 2,3-dihydroxybenzoic acid (2,3-DHBA). Induction of high-concentration [K+]o (20, 70 and 140 mM) significantly increased formation of .OH trapped as 2,3-DHBA in a concentration dependent manner. However, the application of L-NAME (50 mg/kg, i.v.) and allopurinol, a xanthine oxidase inhibitor, abolished the [K+]o depolarization induced .OH generation. Tyramine (1.0 mM) increased the level of 2,3-DHBA. However, the application of L-NAME did not change the level of 2,3-DHBA. On the other hand, pretreatment with allopurinol (10 mg/kg, i.v.) abolished the KCl- or tyramine-induced .OH generation. Moreover, when iron (II) was administered to [K+]o (70 mM)-pretreated animals, there was a marked increased in the level of 2,3-DHBA. However, the application of L-NAME was not related to a Fenton-type reaction via [K+]o depolarization-induced .OH generation. To examine the effect of L-NAME on ischemic/reperfused rat myocardium, the heart was subjected to myocardial ischemia for 15 min by occlusion by left anterior descending coronary artery branch (LAD). When the heart was reperfused, a marked elevation of the level of 2,3-DHBA was observed. However, L-NAME attenuated .OH generation by ischemic/reperfused rat heart. These results suggest that NOS inhibition is associated with a cardioprotective effect due to the suppression of [K+]o depolarization-induced .OH generation. PMID- 11485041 TI - Effects of the cannabimimetic fatty acid derivatives 2-arachidonoylglycerol, anandamide, palmitoylethanolamide and methanandamide upon IgE-dependent antigen induced beta-hexosaminidase, serotonin and TNF alpha release from rat RBL-2H3 basophilic leukaemic cells. AB - There are conflicting reports in the literature as to whether palmitoylethanolamide affects the function of mast cell-related cell lines in vitro, in contrast to the well-documented effects of this compound upon mast cell function in vivo. In the present study, we have reinvestigated the effects of palmitoylethanolamide upon antigen-induced release of [3H]serotonin and beta hexosaminidase from rat basophilic leukemia RBL-2H3 cells and compared these effects with those of 2-arachidonoylglycerol, anandamide and R1-methanandamide. RBL-2H3 cells were sensitized with a monoclonal anti-DNP IgE, after which they were stimulated with antigen (DNP-HSA). Palmitoylethanolamide produced a small, but significant reduction in antigen-stimulated [3H]serotonin release at high concentrations, whereas anandamide was without effect. In contrast, 2 arachidonoylglycerol and methanandamide increased the antigen-stimulated release of both [3H]serotonin and beta-hexosaminidase. It is concluded that in RBL-2H3 cells, these cannabimimetic fatty acid derivatives do not have potent stabilizing effects upon antigen-induced degranulation. PMID- 11485042 TI - Differential effects of trihexyphenidyl on place preference conditioning and locomotor stimulant activity of cocaine and methamphetamine. AB - Cocaine produces rewarding and locomotor stimulant effects by increasing extracellular dopamine (DA) levels in the terminal areas of the mesolimbic DA system. Our recent in vitro studies have shown that a muscarinic receptor antagonist, trihexyphenidyl (THP) inhibits the binding of a cocaine analogue to the DA transporter at concentrations that are ineffective in inhibiting 3H-DA uptake, suggesting that THP may attenuate the actions of cocaine selectively. The present study examined whether THP could affect conditioned place preference (CPP) for and locomotor stimulant activity of cocaine and methamphetamine (MAP) in mice. Mice were injected with cocaine (10 mg/kg) or MAP (1 mg/kg) in one compartment of the CPP chamber 4 times every second day. On alternate days the animals received saline in the other compartment of the CPP chamber. Pretreatment with THP was made 10 min before cocaine or MAP injection. The CPP score and locomotor activity were assessed using a novel activity monitor, SCANET. Cocaine and MAP produced CPP for the drug-paired compartment. Pretreatment with THP (0.05 5 mg/kg) had no influence on cocaine-induced CPP at any dose tested. In contrast, MAP-induced CPP was completely antagonized by THP at 5 mg/kg, which produced no CPP by itself. Another muscarinic receptor antagonist, scopolamine (SCP, 3 mg/kg) neither caused CPP by itself nor affected the development of cocaine- or MAP induced CPP. Both THP and SCP enhanced spontaneous, cocaine- or MAP-induced locomotor activity. Though the present conditioning treatments failed to develop locomotor sensitization to cocaine, THP, but not SCP, acted cooperatively with cocaine to develop locomotor sensitization. The development of locomotor sensitization to MAP was retarded by SCP but was not affected by THP. These results suggest that, contrary to our anticipation, THP has a unique characteristic of specifically counter-acting the rewarding properties of MAP via a non-cholinergic (muscarinic) mechanism. PMID- 11485044 TI - Dibenzylamine--a novel blocker of the voltage-dependent K+ current in myocardial mouse cells. AB - Ventricular myocytes of the mouse ventricle were voltage clamped with a patch clamp technique in the whole-cell configuration. At depolarizing voltage pulses, these myocytes develop a large voltage-dependent K+ outward current. Application of the drug dibenzylamine (DBA) to the bath solution blocked the voltage dependent K+ current. The concentration/response relationship for the peak current at +40 mV indicates a 1:1 binding of the drug to the receptor with a concentration of half maximum effect of 43.1 micromol/l. The block did not require activation of the channels by depolarizing pulses. At concentrations causing partial block (25 micromol/l), the block was independent of voltage. At the same concentration, DBA completely blocked the slow component of the recovery from inactivation (-80 mV) whereas steady-state inactivation was not altered. It is concluded that DBA is a novel blocker of the voltage-dependent K+ current in mouse cardiac myocytes which preferentially affects the current component generating the slow recovery from inactivation. PMID- 11485043 TI - Effects of a novel, selective, sigma1-ligand, MS-377, on phencyclidine-induced behaviour. AB - Phencyclidine (PCP)-induced head-weaving is inhibited by a novel selective sigma1 ligand, (R)-(+)-1-(4-chlorophenyl)-3-[4-(2-methoxyethyl)piperazin-1-yl]methyl-2 pyrrolidinone L-tartrate (MS-377), but not by dopamine D2 antagonists. In the present study, we examined the effects of two potent and selective sigma1 ligands, MS-377 and N,N-dipropyl-2-(4-methoxy-3-(2-phenylethoxy)phenyl) ethylamine (NE-100), on PCP-induced rearing behaviour, hyperlocomotion and ataxia in comparison with the currently available antipsychotic agents with affinity for D2 receptors, haloperidol, sultopride and risperidone. Male Wistar rats or ddY mice were administered MS-377, NE-100, haloperidol, sultopride or risperidone, and PCP was administered 60 min later (in the case of NE-100 10 min later). Rearing behaviour, hyperlocomotion and ataxia were examined 10 min after PCP administration. MS-377, haloperidol, sultopride and risperidone dose-dependently inhibited PCP-induced rearing and hyperlocomotion, but did not antagonize PCP induced ataxia. In contrast, the other selective sigma1-ligand, NE-100, did not affect any of the PCP-induced behaviour patterns in this study. These results suggest that there are at least two types of ligands for sigma1-receptors and that some sigma1-ligands, including MS-377, have more comprehensive effects against PCP-induced abnormal behaviour than other sigma1-ligands or D2 antagonists. PMID- 11485045 TI - Emerging drugs--molecular mechanisms of action of Asian medicines. PMID- 11485046 TI - Structural chemistry and biological activities of natural products from Chinese herbal medicines. AB - This is a brief account of recent developments (1978-1994) in natural product chemistry and bioactivity related to Chinese herbal medicine. In the period covered, more than thousand chemically new constituents were isolated and their structures elucidated. We have presented most of the structural formulae for the sake of clarity, along with passing mentions of pertinent biological activities. The isolates were grouped under different headings according to structure types. PMID- 11485047 TI - Regulation of gene expression by herbal medicines--a new paradigm of gene therapy for multifocal abnormalities of genes. AB - Japanese herbal medicine has long been considered as only supplementary therapy to Western medicine. However, we discovered that an herbal mixture, Saiko-keishi to-ka shakuyaku (SK, TJ-960), showed regulatory function of gene expression such as increased expression of seizure-related gene PTZ-17, proto-oncogene c-fos and heat shock protein HSP 72. These results provide a scientific basis for an important ancient concept and usage of herbal mixtures as a "therapy against diseases which will be suffered in the future". Our results also give an adequate provide break-throughs for therapy and even prevention of intractable epilepsy, Alzheimer's disease, developmental disorders during pregnancy and the postnatal period, and also probably for prevention of metastasis or relapse of various cancers. PMID- 11485048 TI - The effect of EGb 761 on the doxorubicin cardiomyopathy. AB - Doxorubicin (Dx) is used to treat a number of types of cancer. The drug produces many toxic reactions and cardiomyopathy. Many drugs have been used to prevent this myocardial damage caused by peroxidative alterations. EGb 761 is being used to prevent arrhythmias in ischemic myocardium. We decided to establish the tissue protective effect of EGb 761 against myocardial toxic effects of Dx in three groups of rats. Cardiotoxicity signs of Dx were found to be dose-related, beginning at 30 mg/kg dose and being apparent at 45 mg/kg dose. 48 hr after a single i.v. injection, myocardial tissues showed a marked edema, vacuolization and fragmentation. We compared the changes in heart tissue biochemically and histopathologically among the control, Dx treated and EGb 761 (100 mg/kg/d.x 4, i.p.) + Dx treated groups. Biochemical results of CK-MB and MDA values showed a significant decrease in Dx + EGb 761 group when compared with Dx treated group. Histopathologically, myocardial tissues of Dx + EGb 761 treated group were found to have diminished vacuolization and fragmentation. These results suggest that EGb 761 might have the same therapeutic potential in Dx related cardiomyopathy. PMID- 11485049 TI - Protective effect of aminoguanidine against cardiovascular toxicity of chronic doxorubicin treatment in rats. AB - The cardiotoxicity-induced by chronic treatment of doxorubicin have recently be attributed to free radical formation and/or release of nitric oxide. In the present study, an already established rat model of doxorubicin -induced cardiotoxicity was used. Doxorubicin in a total cumulative dose of 15 mgkg(-1) I.P. given in six equal injections over two week period was administered. After three weeks of doxorubicin administration, the blood pressure, serum lactate dehydrogenase, lipid peroxides, asites fluid and mortality rate were significantly increased. Doxorubicin-induced cardiotoxicity was further confirmed by examining the histopathology of heart sections. Myocardial fibres necrosis with prominent acute inflammatory cells were observed in rats hearts treated with doxorubicin. Aminoguanidine, an inhibitor of nitric oxide synthase, 100 mgkg(-1) injected every other day for two week was given concurrently with doxorubicin. Aminoguanidine given concurrently with doxorubicin return blood pressure, lactate dehydrogenase and lipid peroxides to normal control values. Furthermore, aminoguanidine reduces the mortality rate, ascites fluid formation- induced by doxorubicin and improved the histopathology of rats hearts treated with doxorubicin. In conclusion, inhibition of nitric oxide formation may be beneficial in protecting rat hearts against doxorubicin- induced cardiotoxicity. PMID- 11485050 TI - Relationship between hydrophobicity and beta-blocking potencies, affinities or dissociation of beta-blockers from beta-adrenoceptors. AB - The present study was performed to assess the relationship between the hydrophobicity of drugs and (1) inhibitory strength (pA2) on chronotropic or inotropic actions, (2) displacemental potencies of 3H-CGP12177 or 125I iodocyanopindolol binding to beta-adrenoceptors (beta-ARs) (pKi) or (3) dissociating potencies of these drugs from beta-ARs of atria strips pretreated with drugs. The beta-blockers used in the present study were bopindolol, active metabolite of bopindolol (18-502), atenolol, propranolol, pindolol, nadolol, alprenolol, oxprenolol, metoprolol, labetalol and acebutolol. The value of the partition coefficient of propranolol was the highest, and that of the beta1 selective blocker atenolol was the lowest. Although low correlation coefficients between hydrophobicity and inhibitory beta-blocking potencies determined by pharmacological experiments or displacemental potencies by the radioligand binding assay using 3H-CGP12177 and 125I-iodocyanopindolol were observed, significant relationships between hydrophobicities of these drugs and dissociating potencies from beta-ARs were observed. These results suggest that the hydrophobicity of drugs may be important for the slow dissociation from beta ARs, but not for the beta-blocking action. PMID- 11485051 TI - Cadmium accumulation augments contraction and phosphoinositide hydrolysis of vascular smooth muscles. AB - Effects of cadmium accumulation in blood vessels on vasoconstriction and hydrolysis of phosphoinositides were investigated using aortas isolated from rats. The aortic strips were incubated with cadmium chloride (10 microM) for 24 hr and then washed out (cadmium accumulation). In the cadmium-accumulated aortas, contractile responses to KCl and 5-hydroxytryptamine were augmented compared to the control incubated with a vehicle. The degree of the augmentation by cadmium was higher in 5-hydroxytryptamine contraction than in KCl contraction. The level of 5-hydroxytryptamine-induced contraction relative to KCl-induced contraction (expressed as percent of maximum KCl contraction in each strip) was also significantly increased in the cadmium-accumulated aortas as compared to the control. Cadmium pretreatment augmented 5-hydroxytryptamine-stimulated inositol monophosphate accumulation, while there was no difference in the basal level of inositol monophosphate between the cadmium-accumulated aortas and the control. These results suggest that cadmium accumulation increases vasocontractility, in part because of the facilitation of phosphoinositide hydrolysis. PMID- 11485052 TI - An investigation of the mechanisms responsible for acute fluoxetine-induced anxiogenic-like effects in mice. AB - Although selective 5-hydroxytryptamine (5-HT) reuptake inhibitors (SSRIs) are widely used in the chronic treatment of several anxiety disorders, increased anxiety has been observed in some patients at the beginning of treatment with these compounds. Similar increases in anxiety-related behaviors have been observed in animal studies following a single injection with SSRIs. The mechanism underlying this effect is unclear. The aim of the present study was to investigate the effects of a variety of psychoactive compounds on the anxiogenic like activity of fluoxetine. The drugs used included the benzodiazepine diazepam, the 5-HT1A receptor partial agonist buspirone, the 5-HT1A receptor antagonists pindolol and WAY-100635, the non-selective 5-HT2 receptor antagonists methiothepin, mianserin and ritanserin, the non-selective dopamine (DA) receptor antagonist haloperidol, the D1 antagonist SCH23390, the selective D2 antagonist raclopride, the D2/3 agonist quinelorane, the cholecystokininB (CCK(B)) receptor antagonist LY 288513, and the corticotropin-releasing factor1 (CRF1) receptor antagonist CP-154,526. Experiments were performed in the free-exploration test. This model is based on the strong neophobic reactions exhibited by BALB/c mice when confronted simultaneously with a familiar and a novel environment. When administered alone, diazepam (1 and 2 mg/kg), buspirone (1 mg/kg) and mianserin (0.3 mg/kg) produced anxiolytic-like effects as they significantly increased exploratory activity of the novel compartment. In contrast, fluoxetine (20 mg/kg) almost completely suppressed exploration of the novel area. Diazepam reversed the anxiogenic-like as well as the locomotor impairment induced by fluoxetine, while quinelorane blocked only the anxiogenic action of fluoxetine. None of the other compounds was able to counteract this effect. Taken together, these results suggest that dopaminergic mechanisms may underlie, at least in part, the behavioral effects of fluoxetine in the free-exploration test, whereas 5-HT1A 5 HT2, CCK(B) and CRF1 receptors may not be involved primarily in these effects. PMID- 11485053 TI - The role of medial prefrontal cortical dopamine in spontaneous flexibility in the rat. AB - In rat studies, both lesions in the medial prefrontal cortex (mPFC) and alterations of the level of mPFC dopamine (DA) have been found to induce disturbances in behavioural flexibility, as measured with switching tasks. It is not clear whether mPFC DA is also involved in spontaneous flexibility. Therefore, the aim of the present study was to investigate the role of mPFC DA in spontaneous flexibility. As a measure for spontaneous flexibility, the diversity in spatial distribution of exploration on a large open field was used. The rats received local injections into the mPFC with a D1 or D2 antagonist, or the dopamimetic, amphetamine. The results showed that both DA antagonists reduced spontaneous flexibility, due to increased stimulus-bound behaviour. Amphetamine had a similar effect to the DA antagonists. It is suggested that this is most likely due to an amphetamine-induced increase in extracellular DA, leading to a suboptimal level of mPFC DA. PMID- 11485054 TI - An investigation into the acute nootropic effects of Hypericum perforatum L. (St. John's Wort) in healthy human volunteers. AB - Hypericum perforatum L. (St. John's Wort) is a complex herb that has been used for centuries for its putative medicinal properties, and has current therapeutic relevance as a treatment of mild to moderate depression. Recently, two studies in rodents have suggested that hypericum may also have memory-enhancing effects. It has a complex pharmacology, in that acute administration modulates numerous neurotransmitter systems that have previously been observed to either augment or impair a variety of memory processes in humans. This study aimed to examine whether acute administration of standardized hypericum extract could exert a nootropic effect in normal human subjects. The study employed a double-blind, crossover, repeated-measures design. Twelve healthy young subjects completed the Cognitive Drug Research (CDR) memory battery, following administration of placebo, 900 mg and 1800 mg hypericum (Blackmore's Hyperiforte). The findings suggested that hypericum does not have an acute nootropic effect in healthy humans at these doses. However, there was some evidence for an impairing effect on accuracy of numeric working memory and delayed picture recognition at the higher dose. This observed impairment could be due to a sensitivity of these specific tasks to modulation by neurotransmitters that have been noted to have memory-impairing effects (e.g. y-aminobutyric acid (GABA), serotonin). PMID- 11485055 TI - Smoking expectancy mediates the conditioned responses to arbitrary smoking cues. AB - Smokers' responses to smoking cues may be the result of a classical conditioning process. There is evidence that classical conditioning may not proceed in human subjects unless subjects are consciously aware of the stimulus (CS)-reinforcer (UCS) contingencies. In two experiments the role of CS<-->UCS contingency awareness in the expression of conditioned responses (craving, salivation, and skin conductance) was studied. A discriminative classical conditioning paradigm was used during which subjects were presented with one stimulus (the CS +) always paired with cigarette smoking (the UCS) and another (the CS -) never paired with cigarette smoking. Half of the subjects were given instructions to discover the CS<-->UCS contingencies (group 'aware', AWR), whereas the other half were not (group 'unaware', UWR). In experiment 1, all subjects responded to the CS + with increased cigarette craving relative to the CS -; this effect was more pronounced in the AWR group compared to the UWR group. A lower amount of salivation in response to the CS + compared to the CS - was found in the UWR group. These between-group differences were interpreted as a consequence of the enhanced expectancies of smoking in the presence of CS + in group AWR compared to group UWR. In experiment 2, the observed craving responses to CS + and CS - were consistent with those seen in experiment 1, but no discriminative salivary response to the stimuli was found. When, after conditioning training, subjects' expectancies of smoking were removed by instructions, and their responses to CS + and CS - were again measured, the discriminative craving response to CS + and CS was eliminated and all subjects demonstrated a lower amount of salivation in response to the CS + compared to the CS -. These data suggest that presentation of arbitrary cues previously paired with cigarette smoking can elicit CRs and that facilitation of awareness of the CS<-->UCS contingency by instructions can potentiate craving CRs. In addition, these data suggest that craving CRs can be eliminated, whereas compensatory CRs can be facilitated, when cigarette expectancy is removed with instructions. PMID- 11485056 TI - Discrimination of pentobarbital doses and drug mixtures under fixed-ratio and fixed-interval reinforcement schedules. AB - Pigeons were trained to discriminate among 5 mg/kg pentobarbital, 10mg/kg pentobarbital, and saline, under either fixed-interval (FI) or fixed-ratio (FR) reinforcement schedules. When baseline responding stabilized, a higher percentage of responses occurred on the key that produced the reinforcer under the FR schedule than under the FI schedule. After low doses of pentobarbital, responding shifted from the saline key to the 5 mg/kg pentobarbital key; at higher doses of pentobarbital responding shifted to the 10mg/kg pentobarbital key under both schedules. After low doses of ethanol and chlordiazepoxide, responding shifted from the saline key to the 5 mg/kg pentobarbital key, but after high doses of these drugs, responding continued to occur on the 5 mg/kg pentobarbital key under both reinforcement schedules. A 5 mg/kg dose of pentobarbital increased responding on the 10 mg/kg pentobarbital key when it was combined with pentobarbital, ethanol or chlordiazepoxide. Phencyclidine and D-amphetamine produced responding largely on the saline key under both reinforcement schedules. Under the FR schedule, pentobarbital dose-response curves were usually quantal, whereas under the FI schedule the pentobarbital dose-response curves usually were graded. PMID- 11485057 TI - Excitatory mechanisms in neuroleptic-induced vacuous chewing movements (VCMs): possible involvement of calcium and nitric oxide. AB - Tardive dyskinesia (TD) is a serious motor side-effect of chronic neuroleptic therapy. Chronic treatment with neuroleptics leads to the development of oral abnormal movements in rats known as vacuous chewing movements (VCMs). Vacuous chewing movements in rats have been widely accepted as an animal model of tardive dyskinesia. Chronic blockade of D2 inhibitory dopamine (DA) receptors localized on glutamatergic terminals in the striatum leads to the persistent enhanced release of glutamate that kills the striatal output neurons. The object of the present study was to explore the role of glutamatergic modulation on the neuroleptic-induced VCMs. Rats were chronically (for 21 days) treated with haloperidol (1.5 mg/kg, i.p.) to produce VCMs. The neuroleptic-induced VCMs viz., vertical jaw movements, tongue protrusions and bursts of jaw tremors, were counted during a 5 min observation period. Dizocilpine, a non-competitive N methyl-D-aspartate (NMDA) receptor antagonist, dose dependently (0.02 and 0.05 mg/kg) reduced haloperidol-induced VCMs. Felodipine (5 and 10 mg/kg), an L-type calcium-channel blocker, also significantly reduced the VCM count. N-omega-nitro L-arginine methyl ester (L-NAME) (25 and 50 mg/kg), a nitric oxide synthase inhibitor, also reduced the VCM count in an L-arginine-sensitive manner. In conclusion, the findings of the present study indicated NMDA receptor involvement in haloperidol-induced VCMs, and also suggested the possible involvement of calcium and nitric oxide in haloperidol-induced VCMs. PMID- 11485058 TI - Training in the step-down inhibitory avoidance task time-dependently increases cAMP-dependent protein kinase activity in the entorhinal cortex. AB - The cAMP/cAMP-dependent protein kinase (PKA) signaling pathway has been implicated in synaptic plasticity changes and memory consolidation. Several cortical structures are involved in the consolidation of memory for inhibitory avoidance. The aim of the present work was to observe the effects of training in the inhibitory avoidance task on the levels of PKA activity in the entorhinal, parietal and posterior cingulate cortex (EC, PARIET and PC), and the medial precentral area (Fr2) of the rat, at different post-training times (0, 1.5, 3 and 6h). PKA activity, assayed using [gamma-32P]ATP and kemptide, a selective substrate, increased in the EC 3 h after training, but no changes were observed in PARIET, PC and Fr2. These results suggest that the late phase of memory consolidation of inhibitory avoidance requires a functional PKA signaling pathway in the EC in a way that a 'peak' of PKA activity is observed. PMID- 11485059 TI - Development of tolerance to the antiaggressive effects of morphine. AB - Many reports have demonstrated that there is a development of tolerance to many effects produced by morphine. This study was conducted with the aim of determining whether the antiaggressive actions of morphine develop tolerance after chronic administration. Acute morphine administration produced antiaggressive effects which disappeared after chronic (7 days) treatment in isolated mice. An increase in non-social exploration was observed, representing morphine-induced hyperactivity, after acute treatment, which was not present after chronic administration. In conclusion, there is a development of tolerance to the antiaggressive and motor effects of morphine. PMID- 11485060 TI - Processing of grammatical gender in normal and aphasic speakers of Russian. AB - Sensitivity to grammatical gender was investigated in 22 Russian-speaking aphasic patients, compared with young controls. Experiment 1 used a cued shadowing paradigm to assess gender priming (facilitation and/or inhibition of lexical access by a prenominal modifier with congruent, incongruent or neutral gender). Experiment 2 used a grammaticality judgment paradigm with similar stimuli. Normals showed significant interactions between gender and priming in Experiment 1 (facilitation for feminine and neuter nouns but not for masculines) and Experiment 2 (larger effects of context on feminine and neuter nouns) that we interpret as a Markedness Effect. Patients showed significant priming in Experiment 1 and above-chance accuracy in Experiment 2, but failed to show reduced effects for the least-marked masculine gender (the Markedness Effect) in either experiment. Context effects were not related to specific aphasic symptoms or subtypes in either experiment. However, canonical correlation revealed differential effects of specific aphasic symptoms on judgment accuracy (false alarms vs. misses). We conclude that knowledge of grammatical gender is spared in Russian aphasics, but gender processing is deviant. A possible model to account for these differences is discussed. PMID- 11485061 TI - The effect of compressed speech on the ability of right-hemisphere-damaged patients to use context. AB - The ability of RHD patients to use context under conditions of increased processing demands was examined. Subjects monitored for words in auditorily presented sentences of three context types-normal, semantically anomalous, and random, at three rates of speech normal, 70% compressed (Experiment 1) and 60% compressed (Experiment 2). Effects of semantics and syntax were found for the RHD and normal groups under the normal rate of speech condition. Using compressed rates of speech, the effect of syntax disappeared, but the effect of semantics remained. Importantly, and contrary to expectations, the RHD group was similar to normals in continuing to demonstrate an effect of semantic context under conditions of increased processing demands. Results are discussed relative to contemporary theories of laterality, based on studies with normals, that suggest that the involvement of the left versus right hemisphere in context use may depend upon the type of contextual information being processed. PMID- 11485062 TI - Gender differences in laterality on a dichotic task: the influence of report strategies. AB - Four experiments were conducted to investigate the influence of report strategies on gender differences in laterality. Ten males and 10 females completed a dichotic consonant-vowels pairs task under four conditions manipulating reporting strategies. Free recall allowed participants to report syllables in any order they chose, whereas order of report control required the report of a pre specified ear first. In focused attention, they reported only one response from a pre-specified ear. Finally, in ABX discrimination, participants were required to indicate whether a binaural probe was one of the stimuli presented dichotically on the same trial. It was hypothesized that improved control of report strategies would increase the likelihood of detecting significant gender differences in laterality. This was confirmed when results showed no significant gender differences in laterality for the free recall and order of report control conditions, whereas focused attention produced marginal gender differences and clearly significant differences were obtained in the ABX discrimination condition. These findings have implications for interpretations of gender differences in laterality based on strategy effects. Interpretations emphasize the role of attention deployment in the measurement of laterality. PMID- 11485063 TI - Selective impairments for addition, subtraction and multiplication. implications for the organisation of arithmetical facts. AB - This study reports for the first time a selective impairment for simple addition in patient FS. Moreover, patient VP presented with a selective impairment for simple multiplication and patient DT with a selective impairment for simple subtraction. These findings are discussed in the context of two of the most influential models for the organisation of arithmetical facts in memory (Dehaene and Cohen, 1995, 1997, and Dagenbach and McCloskey, 1992). Dehaene and Cohen (1995, 1997) have proposed that dissociation between arithmetical facts result from a selective impairment to two different types of processing: rote verbal memory for multiplication and simple addition vs. quantity processing for subtraction and division. Dagenbach and McCloskey (1992) suggest dissociation between arithmetical facts result from a selective damage to segregated memory networks specific for each operation. We will argue that our findings are problematic for Dehaene's model and in good accord with McCloskey's view. PMID- 11485064 TI - Influence of visual distractors on movement trajectory. AB - In the present study, we explored the influence of a visual distractor on the trajectory of movements made to join two dots. The two dots could be unconnected (D) or connected by a straight (L) or curved line, convex either to the left (LL) or to the right (RL). The connecting line constituted the visual distractor. In Experiment 1, subjects were asked to perform the joining movement as straight as possible. The results showed that hand trajectory moved to the left of the midsagittal axis in LL and to the right in RL, while it was almost straight both in L and D. In Experiment 2, subjects were explicitly required to follow the connecting line during their movements to verify whether, in the previous experiment, they had used curved lines as guide for their movements. An increase of movement time and different hand paths showed that this was not the case. In Experiment 3, subjects were asked to move as straight as possible without vision of their hand. Hand trajectories were shifted to the left in all experimental conditions, but the leftward shift was greater in LL than in all other conditions, and also greater in D and L than in RL. These findings suggest that the visual distractor influenced hand trajectory by attracting subject's attention and competing with target for motor response. It is hypothesized that the attracting influence operated by the distractor was sustained by involuntary attentional mechanisms. PMID- 11485065 TI - Naming of musical notes: a selective deficit in one musical clef. AB - We investigated the ability to perform solfeggio, i.e. oral reading of musical notes in MP, a 65 year-old female professional musician, who, following a left temporoparietal ischemia, showed a complex pattern of amusia. The deficit on which we focused was her inability to read orally the bass (F) clef, often substituting it with the violin (G) clef. This problem could not be attributed to a lack of comprehension. The patient could in fact correctly perform on the piano the same sequences she erroneously read aloud; she was also able to correctly judge whether two strings, one in bass clef and the other in violin clef, represented the same sequence of notes. The problem seems to lie in the inability to retrieve note names keeping into account the clef-rule. It is hypothesized that, in the production of note names, this function requires the identification and application of syntactic-like information, in analogy with what is thought to happen in the retrieval of other words. PMID- 11485066 TI - A longitudinal assessment of diverging verbal and non-verbal abilities in the Williams syndrome phenotype. AB - Jarrold et al. (1998) presented evidence to suggest that verbal and non-verbal abilities develop at different rates in individuals with the Williams syndrome phenotype. However, this evidence was derived from cross-sectional rather than longitudinal data. The current report presents data from a series of follow up assessments which examine the development of vocabulary and pattern construction abilities in 15 of the original sample of 16 individuals, over a 40 month period. The results confirm the original predictions, as mental age equivalent scores for vocabulary increase more rapidly than scores for the pattern construction test; a finding, which appears unlikely to be due to practice effects. PMID- 11485067 TI - Finger movement tracking control in 8- and 9-year-old males and females in stimulus-response compatible and non-compatible hand positions. AB - This study examined whether differences exist between 8-9 year-old males and females in two finger movement tracking tasks with different degrees of stimulus response (S-R) compatibility. Sixteen males and 20 females performed finger extension and flexion movements to track a computer screen cursor along a sine wave target in two different hand positions. Subjects had control of cursor movement only in the vertical plane as the cursor moved automatically from left to right across the target. In the compatible position, the hand was pronated so that finger motion occurred in the vertical plane. In the non-compatible position, the hand was neutral between pronation and supination so that finger motion occurred in the horizontal plane. The results showed that males tracked significantly more accurately than females in both the compatible and non compatible positions. Also, an S-R compatibility effect was found in females but not in males. These results are consistent with other reports of higher information processing performance in adult males compared to females during visual-spatial tasks. The results are discussed as a possible manifestation of hormonally induced brain organization differences between males and females. PMID- 11485068 TI - AIDS vaccines: money games and elusive strategies. PMID- 11485069 TI - Detection and molecular characterization of the zot gene in Vibrio cholerae and V. alginolyticus isolates. AB - A total of 11 Vibrio cholerae isolates from 1996-1998 outbreaks in Malaysia and 4 V. alginolyticus were analyzed. Isolates were characterized by polymerase chain reaction (PCR) and Southern hybridization for the presence of the gene encoding zonula occludens toxin (zot). Screening of zot gene by PCR revealed the presence of this gene in V. cholerae and V. alginolyticus. The zot gene from one V. cholerae Ogawa isolate that was cloned in a pCR 2.1 TOPO vector was sequenced. The sequences obtained were 99% homologous to the zot gene sequence from the Gene Bank. PMID- 11485070 TI - Development of duplex PCR assay for rapid detection of enterotoxigenic isolates of Clostridium perfringens. AB - A duplex PCR assay was developed for the rapid and specific amplification of the alpha-toxin (phospholipase C, plc) gene and the enterotoxin (cpe) gene from Clostridium perfringens. Two pairs of primers were newly designed for the species identification and also for the differentiation between enterotoxin-positive and enterotoxin-negative C. perfringens strains in a single reaction. The detection by agarose gel electrophoresis yielded 2 bands of 280-bp of plc and 420-bp of cpe for all four enterotoxin-positive reference strains tested without the need for further hybridization, and one band of 280-bp of plc for all seven enterotoxin negative reference strains. While 50 strains of other Clostridium species and other bacteria tested by PCR were negative for both genes. The detection limit, as measured with purified DNA was 10 fg or as few as 4 organisms could be detected. This assay was used to identify primary fecal spore isolates from 244 fecal specimens of patients with diarrhea. Of total 432 colonies from 144 positive growth cultures determined, 21 revealed both plc and cpe genes and 411 were positive for plc gene only. This suggested a prevalent of 5% of all C. perfringens strains that carry the enterotoxin gene. The results indicate the duplex PCR as a simple, sensitive, specific, cost-effective and time saving assay for detection of potentially enterotoxigenic isolates of C. perfringens, and has potential application for epidemiological investigations of food poisoning outbreaks and quality control of food products for humans and animal feeds. PMID- 11485071 TI - Detection of Mycobacterium tuberculosis from sputum specimens using one-tube nested PCR. AB - One-tube nested PCR was developed for diagnosis of pulmonary tuberculosis using sequences based on thel6SrRNA gene. The usage of primers 16SOL, 16SOR, 16SIL and 16SIR with optimized conditions could detect 555 bp DNA band from 21 species, 41 strains of mycobacteria and one isolate of Nocardia asteroides. It also revealed a specific 306 bp DNA band from 59 strains of M. tuberculosis complex. Cross amplification was observed in M. marinum, M. ulcerans and a few isolates of M. fortuitum complex. The developed method could detect as little as 100 fg of M. tuberculosis DNA. The PCR mixtures could be stored at 0 degrees C for 2 months or at -20 degrees C for at least 20 months without decrease in sensitivity. Using one-tube nested PCR for detection of M. tuberculosis compared with acid fast staining and culture results from 153 sputum specimens revealed 88.6% sensitivity and 89.2% specificity in smear positive specimens and 93.2% sensitivity and 85.0% specificity in culture positive specimens. PMID- 11485072 TI - Invasive pneumococcal infection in children. AB - Streptococcus pneumoniae infections can involve multiple organs and cause high mortality and morbidity. In this retrospective study, we reviewed patients with invasive pneumococcal infection in the pediatric department of a teaching hospital in Taipei. From 1984 to 1998, 81 children with invasive pneumococcal infection were hospitalized. Twenty-eight patients had meningitis, 27 had pneumonia with pleural effusion, 60 had sepsis, and 4 had arthritis. Thirty-eight patients had more than one site of infection. Most of our patients (81.7%) were below 5 years of age. Pneumococcal infections were more common from October to March. Eight patients had a history of trauma that correlated with the site of infection. Thirteen patients (16.0%) expired and 20 (24.7%) had severe sequelae. Multi-regression analysis found that meningitis and complications were independent variables that affected the outcome. The percentage of penicillin resistant strains increased beginning in 1990 and accounted for about four-fifths of the infections in the final 2 years of the study. Since invasive pneumococcal infections in children may have a poor prognosis and penicillin-resistant strains have become increasingly common, early and adequate antibiotic therapy should be given as soon as possible. PMID- 11485073 TI - Evaluation of a newly developed dipstick test for the rapid diagnosis of scrub typhus in febrile patients. AB - Scrub typhus is a potentially fatal, febrile disease prevalent in rural Asia. The etiological agent, Orientia tsutsugamushi, is transmitted to humans by the bite of a larval trombiculid mite. No current diagnostic test is sufficiently practical for use by physicians working in rural areas. A new dipstick test using a dot blot immunoassay format has been developed for the serodiagnosis of scrub typhus. We evaluated this test on 83 patients presenting with acute fever of unknown origin at Maharaj Hospital, a tertiary care medical center in Nakhon Ratchasima, Northeast Thailand. The diagnosis of scrub typhus was confirmed in 30 of these patients (36%) by the indirect immunoperoxidase test. The sensitivity of the test was 87% and its specificity was 94%. The dot blot immunoassay dipstick is accurate, rapid, easy to use, and relatively inexpensive. It appears to be the best currently available test for diagnosing scrub typhus in rural areas where this disease predominates. PMID- 11485074 TI - Retrospective analysis of epidemiological investigation of Japanese encephalitis outbreak occurred in Rourkela, Orissa, India. AB - There was an outbreak of Japanese encephalitis (JE) during July/August of 1993 in Rourkela City of Orissa, India. Among the serum samples, 40% of the sera showed antibodies against JE, while 17% of the sera showed recent infection to dengue virus. As many as 15 species of mosquitos were encountered in and around the affected areas. Species like Culex tritaeniorhynchus, Cx, vishnui, Cx. pseudovishnui and Cx. bitaeniorhynchus constituted 19% of the total collections. The above species were resistant to DDT and dieldrin, but susceptible to malathion. PMID- 11485075 TI - Impact of intestinal helminth infection on anemia and iron status during pregnancy: a community based study in Indonesia. AB - A cohort study was carried out in Purworejo District, Central Java, Indonesia to investigate prevalence of anemia and low iron stores during pregnancy in relation to intestinal helminth infection. 442 pregnant women were followed until 5-10 weeks postpartum, during the period of April 1996 - August 1998. Information on intestinal helminths, hemoglobin and serum ferritin was collected each trimester. Highest prevalence of anemia in pregnancy (37.1%) was found in the second trimester, while the highest prevalence of low iron stores (49.5%) was found in the third trimester. Most pregnant women (69.7%) were infected with at least one species of pathogenic intestinal helminths. The most common helminth detected was Trichuris trichiura followed by Necator americanus (hookworm) and Ascaris lumbricoides. A significant negative association was found between hookworm infection and serum ferritin at the first trimester (p = 0.010). It is concluded that hookworm infection can interfere iron stores. Other causes of anemia among pregnant women should be considered. Anthelminthic therapy could be given to infected women before conception as public health strategy to improve iron status. PMID- 11485076 TI - A comparative study of Phyllanthus amarus compound and interferon in the treatment of chronic viral hepatitis B. AB - Fifty-five patients with chronic viral hepatitis B were randomly divided into two groups. Thirty patients were treated with Phyllanthus amarus compound (PA Co) for three months in the treatment group, another 25 patients were treated with domestic recombinant human interferon alpha-1b (IFN-alpha 1b) for three months as controls. The total effective rate in the treatment group was 83.3%, showing no significant difference from the control (p>0.05). The normalization rates of ALT, A/G and SB in the treatment group were 73.3%, 80.0% and 78.2% respectively, which were significantly higher than that in the control (p<0.05). The negative conversion rates of HBeAg and HBV-DNA in the treatment group were 42.3% and 47.8%, showing no significant difference from the control (p>0.005). It is indicated that PA Co has remarkable effect for chronic viral hepatitis B in recovery of liver function and inhibition of the replication of HBV. PMID- 11485077 TI - Seroprevalence of anti-human herpes virus-6 IgG antibody in children of Bangkok, Thailand. AB - The prevalence of HHV-6 infection was surveyed by determining the presence of anti-human herpesvirus-6 IgG (Anti-HHV-6 IgG) using an ELISA method. Two hundred and ten sera collected from healthy Thai children aged between 0 to 12 years (mean +/- standard deviation = 3.35+/-3.33) indicated the prevalence of HHV-6 infection was 88.10% (185/210). Samples were classified into 7 groups, 30 samples each, according to their ages, ie, group 1: 0 - < 6 months; group 2: 6 - < 12 months; group 3: 12 - < 18 months; group 4; 18 - < 24 months; group 5: 2 - < 5 years; group 6: 5 - < 8 years and group 7: 8-12 years. The prevalence of HHV-6 infection was 63.33%, 70%, 96.67%, 93.33%, 100%, 100% and 93.33%, respectively. The mean level of anti-HHV-6 IgG among those positive for HHV-6 infection (185 samples) increased from 0 < 6 months old (17.47+/-6.32 units) to 27.57+/-8.42 units in 6 - < 12 months old, with the highest value found in the 18 - < 24 months old group (33.08+/-8.64 units). The level declined thereafter. A statistically significant difference of the mean level of anti-HHV-6 IgG among positive groups was found (p-value < 0.05). The important factor associated with HHV-6 infection was age (p = 0.002), while sex, socioeconomic status, number of children in the family and child rearing place did not show any association. PMID- 11485079 TI - Rapid diagnosis of cytomegalovirus infection in congenital neonates. AB - Immunostaining was compared with PCR for diagnosis of congenital CMV infection. IgM and IgG antibody assays were also performed in parallel. Immunostaining gave sensitivity and specificity of 60% and 97% respectively. Correlations among immunostaining, PCR and the presence of IgM antibody was reported. Immunostaining can be used for early diagnosis of congenital CMV infection in parallel with detection of IgM antibody. PMID- 11485078 TI - Prevalence of cytomegalovirus in Thai blood donors by monoclonal staining of blood leukocytes. AB - Four hundred and forty-one blood and serum samples were collected during August to October 1998 from the blood donors at the blood bank of Rajvithi Hospital, Bangkok, Thailand. Their ages were varied between 18-55 years. All specimens were tested by immunostaining and ELISA methods. Forty-seven specimens (10.66%) gave positive results by immunostaining. Among these, 20 cases were seropositive and 27 cases were seronegative. The age group between 41-50 years had a high percentage of CMV infection as judged by the immunostaining method, more than the other age groups. By ELISA, 231 cases (52.38%) had positive IgG antibody to CMV, 42 cases (9.52%) were IgM antibody positive and 39 cases (8.84%) were positive for both IgG and IgM antibodies. The age groups between 36-40 years had a higher percentage of IgM antibody positives than the other age groups. Since the immunostaining method can detect early CMV infection, screening for the presence of antibodies alone is not enough to rule out CMV infection. Immunostaining along with ELISA detection of antibodies was useful for determining a decrease in CMV infection. PMID- 11485080 TI - Bacterial causes of AIDS-associated diarrhea in Thailand. AB - The incidence of bacterial diarrhea in AIDS patients has increased steadily and has led to enormous medical and public health problems. In this study, the clinical data together with 350 rectal swab samples each from AIDS patients with diarrhea (APD) and non-AIDS patients with diarrhea (NAPD), were collected and examined for bacterial enteropathogens at the Bamrasnaradura Infectious Diseases Hospital (BIDH), Nonthaburi, Thailand from May to December 1996. Patients were matched by age and sex. The majority of these patients were male (79%, 554/700), aged between 15 and 34 years (70.9%). The study found that the isolation rates of bacterial enteropathogens causing diarrhea in APD (18%, 62/350) were considerably lower than those in NAPD (43%, 152/350) (p<0.05). The infection rate with Salmonella group B (19.7%, 12/61) in APD was found to be significantly higher than that in NAPD (14.3%, 2/14) (p<0.05). Vibrio parahaemolyticus (53.3%, 81/152), Plesiomonas shigelloides (27%, 41/152), Aeromonas spp (19.1%, 29/152) and V. cholerae O1 (15.1%, 23/152), were more frequently detected in NAPD than in APD (p<0.05). Only nine Escherichia coli strains were isolated from APD, of which six were enteroinvasive E. coli, two enterotoxigenic E. coli and one enterohemorrhagic E. coli (non O157) possessing both vtl and vt2. No V. cholerae strains were detected in APD. The least effective antibiotics were ampicillin, tetracycline and cotrimoxazole. Antibiotic resistant patterns of the isolated organisms were similar from both groups. The results from this study might be useful in Thailand in the diagnosis and management of clinical cases of bacterial diarrhea, especially APD. PMID- 11485081 TI - Cutaneous manifestations in HIV positive patients. AB - Cutaneous manifestations are common clinical findings among HIV positive patients. The causes may be bacteria, viruses, fungi and other non-infectious agents. This study was conducted at the Pramongkutklao Hospital skin clinic to determine the frequency distribution of cutaneous manifestations in HIV positive patients. A total of 147 patients with HIV seropositivity were recruited and divided into a retrospective group and a prospective study group. For the retrospective study, hospital records of 129 patients who attended from January 1995 to November 1998 were recruited. The prospective study was carried out from November 1998 to January 1999 and 18 patients were recruited. Cutaneous finding among patients in the two studies were evaluated. There were ten common cutaneous manifestations observed in the retrospective and prospective study including pruritic papular eruptions (PPE) (51.2%, 50%), oral candidiasis (16.7%, 21.7%), herpes zoster (10.9%, 5.6%), oral hairy leukoplakia (10%, 5.6%), unclassified eczema (9%, 11.1%), urticaria (5.6%, 3.1%), seborrheic dermatitis (4.7%, 16.7%), folliculitis (4.7%, 5.6%), prurigo simplex (4.7%, 5.6%), and Steven-Johnson syndrome (3.9%, 0%). However, the distribution of cutaneous manifestations in the two studies were not significantly different. These findings may be useful as baseline data for common cutaneous manifestations in HIV positive patients. PMID- 11485082 TI - Evaluation of a new fourth-generation microwell enzyme-linked immunosorbent assay for detection of HIV-1 subtype B and E antibodies. AB - The recent fourth-generation enzyme-immunoassays have been used to increase the sensitivity for detecting HIV-1 antibodies and reduce the window period of HIV infection. The HIV antigens utilized in those assays were prepared from HIV-1 clade B which is different from HIV-1 subtypes circulating in Thailand. We evaluated 323 HIV-1 seropositives either B or E subtype to determine whether they were detected with the new combined anti-HIV and the p24 Ag assay. Under evaluation we found that this enzyme immunoassay manufactured by Organon Teknika showed the high sensitivity and specificity with a greater delta (delta) value with B than E subtypes samples (+15.29 vs +5.73). PMID- 11485083 TI - Prenatal diagnosis of hemoglobin Bart's hydrops fetalis by HPLC analysis of hemoglobin in fetal blood samples. AB - Since HbF and HbA are not found in fetuses with Hb Bart's hydrops fetalis the feasibility of prenatal diagnosis of homozygous alpha-thalassemia 1 by fetal hemoglobin typing was examined. Blood samples were obtained from fetuses at 18 to 22 weeks of gestation by cordocentesis in 32 pregnant women at risk of having a child with homozygous alpha-thalassemia 1 (alpha-thal-1). The samples were analyzed by a PCR-based method for the diagnosis of alpha-thal-1 (SEA type) and the proportion of hemoglobin fractions were determined by automated HPLC. DNA analysis showed that 8 of the 32 fetuses were homozygotes for alpha-thal-1, 17 were heterozygous for alpha-thal-1 (alpha-thal-1 trait), and a normal complement of four a-globin genes was found in 7 cases. The Hb typing in fetuses with homozygous alpha-thal-1 showed a peak of unbound Hb (Hb Bart's and Hb Portland) and no HbF, HbA and HbA The alpha-thal-1 trait chromatograms showed unbound Hb, pre HbF, HbF and HbA peaks. The chromatogram of normal fetuses showed HbF and HbA peaks without HbA2. In these cases the HbA proportion is between 3% and 10% with no apparent differences between the 18h and 22nd week of gestation. As the analysis of fetal Hb types by HPLC is facile and speedy and the results correspond with those obtained by DNA analysis, fetal Hb typing by automated HPLC is a convenient prenatal diagnostic method for homozygous alpha-thal-1. The method is recommended for prenatal diagnosis in populations with a high frequency of alpha-thal-1. PMID- 11485084 TI - The prevalence and pattern of wife beating in the Trincomalee district in eastern Sri Lanka. AB - A descriptive cross-sectional study was carried out to determine the prevalence and to identify some socio-demographic factors associated with wife beating in the Medical Officer of Health (MOH) area of Kantale in the Trincomalee district of eastern Sri Lanka. A random sample of 417 women in the age category 18-49 years constituted the sample population. Data were obtained by focus group discussions followed by the administration of a structured questionnaire by trained interviewers. The prevalence of reported wife beating among ever-married women was 30% and the prevalence of wife beating in the year preceding the study was 22%. There was no significant association between wife beating and ethnicity of the study population or a particular age group of either the batterer or the victim. Moreover, wife beating was associated with an early age at marriage for women, low-income, a low standard of living index, large families and alcohol consumption by the batterer. A significant inverse relationship between domestic violence and the level of education of both the batterer and the victim was also identified. Contusions, typically distributed in the region of the head, face and neck were found to be the commonest type of injury suffered by battered women. A majority of women, irrespective of their level of education and employment status placed the welfare of their children as the prime reason for continuing to stay in an abusive relationship. The study concludes that wife beating is a serious health and social problem for the women population of Kantale. Intervention is recommended in relation to key issues identified by the study, including alcohol abuse by men, relative lack of education among the population, lack of family planning, societal influences promoting teenage marriages of the girl-child and absence of programs aimed at creating awareness on wife beating. PMID- 11485085 TI - Physical activity and risk factors for hip fractures in Thai men. AB - To test the hypothesis that hip fracture is associated with physical activity in Thai elderly men, a case-control study was conducted in Bangkok, Thailand. A total of 187 men aged 51 years over, resident in Bangkok, admitted consecutively with a radiologically confirmed first hip fracture were studied. 177 age-matched community controls were randomly recruited from the same neighborhood of the cases. Physical activity was independently associated with reduced risk of hip fracture after controlling for confounding factors. Very active and active past physical activity markedly reduced risk of hip fracture in comparison to subjects with inactive past physical activity. Recent active physical activity was also protective against hip fracture. This prompts a need to identify strategy to promote physical activity among the elderly and at an early age. PMID- 11485086 TI - A preliminary study of the distribution of blood group systems in Thai blood donors determined by the gel test. AB - Two hundred blood samples obtained from volunteer blood donors at the Blood Bank, Army Institute of Pathology were studied for red cell groupings in the ABO, Rh, MNSs, Duffy, Lewis. P. Kell, Lutheran and Kidd Systems. Each sample was tested by the gel test using five cards; the ABO-Rh card, Diaclon Rh sub groups + K card, Antigen profile I card (P, Le(a), Le(b), Lu(a), Lu(b)), Antigen profile II card (k, Kp(a), Kp(b), Jk(a), Jk(b)) and Antigen profile III card (M, N, S, s, Fy(a), Fy(b)). For the ABO System, group O is the most common (40.5%) followed by group B (30.5%), group A (20.5%) and group AB (8.5%). The most common Rh gene complex was CCDee (51.5%), which was similar to other studies. The incidence of MMss and MNss gene complexes were the most common in the MNSs System. Fy(a) is very common as in other Asians. In the Lewis System, the incidence of Le (a-b-) was 23.5%, which is consistent with other findings in the Thai population. Sixty (30%) were positive with anti-P1. For the Kell System, only kk and Kp(b) positive types were observed in this study, as well as Lu (a-b+) in the Lutheran System. Jk (a-b-) was not found, which is considered a rare phenotype among Thai people. This study reveals the blood group distribution in 200 Thai volunteers using the gel test. Because of its simplicity and efficacy, this test is practical in population studies. Moreover, it is useful for mass screening and application in emergency situations. PMID- 11485087 TI - Phase II clinical trial on effect of the long turmeric (Curcuma longa Linn) on healing of peptic ulcer. AB - The study examined patients who had symptoms indicating peptic ulcer. Forty-five patients, 24 males and 21 females, aged between 16-60 years were included in the study. Twenty-five patients, 18 males and 7 females, were endoscoped, their ulcers located in the duodenal bulb and gastric (angulus). The ulcer sizes varied between 0.5 to 1.5 cm in diameter. Capsule-filled turmeric was given orally in the dose of 2 capsules (300 mg each) five times daily, one half to an hour before meals, at 16.00 hours and at bedtime continuously. The result after 4 weeks of treatment showed that ulcers were absent in 48% or 12 cases (DU 9 and GU 3). Eighteen cases (DU 13 and GU 5) had absence of ulcer after 8 weeks of treatment. Nineteen cases (76%) (DU 14 and GU 5) did not have ulcers after 12 weeks of treatment. The rest, 20 cases were not found to have ulcers and some were not endoscoped. They appeared to have erosions, gastritis and dyspepsia. They received turmeric capsules for 4 weeks of treatment. The abdominal pain and discomfort satisfactorily subsided in the first and second week. They could take normal foods instead of soft meals. Blood chemistry and hematology of all 54 patients had no significant changes in hematological system, liver and renal functions both before and after treatment. PMID- 11485088 TI - Stability of ceftazidime in normal saline solution after exposure to light. AB - Bactericidal activity of ceftazidime is determined by the time that concentrations in tissue and serum are above the MIC for the pathogens during the dosing interval. Thus, the most effective mode of administration of ceftazidime is continuous infusion. However, this agent is light sensitive which may result in instability when administered by this method without protection from light. Until now we have had no data to demonstrate the stability of this drug during continuous infusion. Therefore, the objective of this study was to provide such data. One gram of ceftazidime was mixed with 1,000 ml normal saline and exposed to two 36 watt fluorescence lights for 24 hours. The distance between ceftazidime solution and light source was 1 meter. Twenty samples (1 g-ceftazidime in normal saline) solution were evaluated. The mean ceftazidime concentrations in normal saline solution were decreased by only 1.69%, 4.44% and 7.19% after 6, 12 and 24 hours after exposure to light, respectively. Therefore, we conclude that the reduction of drug concentration was not considered to be significantly high, and this agent can be administered by continuous infusion. PMID- 11485089 TI - Geo-helminth infections in a rural area of Sri Lanka. AB - School children carry the heaviest burden of morbidity due to intestinal helminth infection. The objective of this investigation was to study geo-helminth infections in 349 school children aged 6 to 13 years living in a rural area of Sri Lanka. Stool samples were examined by direct saline smear in an initial survey to determine the prevalence of intestinal parasitic infections and thereafter the children were followed up over a two year period with cross sectional surveys of stool samples being carried out at yearly intervals. Following collection of a stool sample, all the subjects were treated with mebendazole 500 mg as a single dose. Weights and heights were measured using standardized procedures. 2 ml of venous blood were collected from each subject under aseptic conditions to determine hematological indices. The prevalence of geo-helminth infections was low, and the prevalence declined during the two-year period from 5.4% in 1997 to 2.2% in 1998 and 2.0% in 1999 following yearly mass anti-helminth treatment. The incidence density was 0.021 cases per child year. The reduction in the prevalence from the baseline to the second survey is probably due to the reduction of the reservoir of infection among children as a result of mass treatment at baseline. The prevalence of infection during the second and third surveys were almost the same probably due to infections originating from other segments of the untreated population. PMID- 11485090 TI - Schistosomiasis malayensis-like infection among the Penan and other interior tribes (Orang Ulu) in upper Rejang River Basin Sarawak Malaysia. AB - A serosurvey of various indigenous interior tribes (Orang Ulu) in upper Rejang River Basin Sarawak Malaysia, the site of a multibillion Ringgit hydroelectric power project, found 6.8% of the individual surveyed were seropositive for schistosomiasis, as determined by ELISA method using the soluble egg antigen of Schistosoma malayensis Baling strain. In all age group, the seroprevalence rate is higher (9.5%) in males than in females (4.5%) except for the 31-40 age group. Seroprevalence of schistosomiasis was found to increase with age with the above 60 age group having the highest rate followed by the 31-40 age group. Seroprevalence rate among the tribes ranges from 4.1% among the Penan to 11.6% among the Kajang. There was no seroevidence of schistosomiasis among the Ukits. A snail survey found four snail species including Brotia species, the intermediate host of the lung fluke Paragonimus westermani, however no schistosome snail host was identified. Although schistosomiasis malayensis-like infection may be endemic in the area, its public health significance remains undetermined. PMID- 11485091 TI - Intestinal microsporidiosis in HIV-infected children with acute and chronic diarrhea. AB - A prospective study of intestinal microsporidiosis in HIV-positive children was conducted at the Queen Sirikit National Institute of Child Health and Phramongkutklao Hospital, Bangkok, Thailand. Hospitalized HIV-positive children with and without diarrhea were enrolled in this study. Microsporidial spores identified by calcofluor fluorescent and gram-chromotrope stain were confirmed by electron microscopy. As well as Cryptosporidium parvum, Microsporidia was the most common protozoa found in the present study, each was 7.1%. Microsporidia was significantly more common in those who had diarrhea. Intestinal microsporidiosis was found in HIV-positive children with both acute and chronic diarrhea. This study emphasizes the importance of Microsporidia in HIV-infected children. Early detection of microsporidia could be of benefit for the patients, since the infection is treatable. PMID- 11485092 TI - A survey of Toxoplasma gondii antibodies in pigs in Indonesia. AB - In August 1994, blood samples were collected from 110 pigs at a slaughter house in Bandar Lampung (Sumatra) and from 98 pigs in a slaughter house in Ujung Pandang (Sulawesi), 208 pigs in total. Anti-Toxoplasma gondii antibody titer in these samples was surveyed using the latex agglutination test. The positivity rate was 3.6% (4/110) in Bandar Lampung, and 9.2% (9/98) in Ujung Pandang. The antibody titer was 1:128 in two and 1:512 in two in Bandar Lampung, and 1:64 in six, 1:128 in two, and 1:256 in one in Ujung Pandang, showing that the titer was higher in Bandar Lampung. PMID- 11485093 TI - Relationship of intestinal parasites to the environment and to behavioral factors in children in the Bolikhamxay Province of Lao PDR. AB - From March to July 1998 the infection rates of 732 children aged below 15 years were assessed. The investigation was conducted in selected villages of the Bolikhamxai Province in Lao PDR. Socio-economic conditions and behavioral pattern were studied. The three soil-transmitted helminths, Ascaris lumbricoides, Trichuris trichiura and hookworm were found with prevalence rates of 67.14, 17.49 and 12.83%, respectively. Infection rates with other intestinal parasites were negligible. Of the children investigated, 56.7% harbored one and 20.45% more than one parasite. Except for hookworms, no statistically significant differences were found between genders. The probability of being infected with A. lumbricoides is associated with living in mountainous areas. For hookworms, infection is associated with staying in the plains. A river in the vicinity of the village is linked with the probability of being infested with Trichuris trichiura. Not to belong to a family with the ability to own expensive items increases the probability by almost two times of getting infested with A. lumbricoides. Unhygienic behavioral factors were important in increasing the probability of suffering from A. lumbricoides and T trichiura infection. Behavioral factors did not seem to be related to hookworm infections. It was concluded that after mass treatment, besides promoting the construction of toilets, it is also important to improve personal hygiene so that a lasting impact on the infection rate of the most prevalent parasites in Lao PDR could be achieved. Measures to control parasitic infections do not have to be postponed until a marked improvement of the economic situation has occurred. PMID- 11485094 TI - Drug resistant malaria on the Thai-Myanmar and Thai-Cambodian borders. AB - We describe the changing epidemiology of drug resistant malaria in Thailand over the past decade. Factors determining the characteristic patterns of the development and spread of resistance to anti-malarial drugs on the Thai-Cambodian border and the Thai-Myanmar border are explored, namely, population dynamics, drug usage and malaria control measures. The introduction of artesunate mefloquine combination in selected areas along the two borders in 1995 is believed to be one of the multiple factors responsible for stabilizing the multidrug resistance problems in Thailand today. Other control measures and inter governmental co-operation must continue to be strengthened in order to limit the spread of drug resistance malaria in the Southeast Asian region. PMID- 11485095 TI - Frequency of early rising parasitemia in falciparum malaria treated with artemisinin derivatives. AB - To define the frequency of the early rising of parasitemia in falciparum malaria patients treated with artemisinin derivatives, a retrospective chart review of 497 patients admitted to the Hospital for Tropical Diseases, Bangkok in 1996 was carried out. Early rising parasitemia, defined as an increase in the parasite count over the baseline pretreatment level during the first 24 hours of treatment, was found in 59/229 episodes (25.8%) of uncomplicated, and 111/268 episodes (41.3%) of complicated falciparum malaria. All uncomplicated cases were successfully treated without developing any complications. There were 2 deaths and 13 changes of drug regimen in the complicated group. Only one of these unfavorable responses was due to parasite response. Early rising parasitemia was very common in falciparum malaria treated with artemisinin derivatives, despite their ability to clear the parasitemia, and did not indicate failure of the drug used. PMID- 11485097 TI - Effects of hydroxypyridinone iron chelators in combination with antimalarial drugs on the in vitro growth of Plasmodium falciparum. AB - Using standard in vitro drug susceptibility methods, we assessed the antimalarial activity of 3 orally administered iron chelators (hydroxypyridinones) alone and in combination with conventional antimalarials drugs (quinine, mefloquine, artesunate, tetracycline, atovaquone) against a chloroquine-resistant Plasmodium falciparum isolate. When tested alone, all iron chelators and antimalarial compounds inhibited the growth of the parasites. IC50 values for iron chelators were 60-70 microM, whereas the IC50 values for antimalarial drugs were in nM ranges, with artesunate being the most potent. The derived isobolograms for the interaction of hydroxypyridinones and antimalarial drugs showed addition or mild antagonism, similar to desferroxamine (Sum of Fractional Inhibitory Concentration, sigma FIC < 0.5 or > 4.0). Despite the absence of synergy with conventional drugs, intrinsic antimalarial activity of hydroxypyridinones supports the continued assessment of these iron chelators as treatment adjuncts. PMID- 11485096 TI - Can treatment of P. vivax lead to a unexpected appearance of falciparum malaria? AB - Of 994 patients admitted to the Bangkok Hospital for Tropical Diseases for P. vivax malaria, 104 (10.5%) experienced appearance of Plasmodiumfalciparum following drug treatment for P. vivax . In all patients, P. falciparum parasites were not found by microscopic examination upon admission. The mean time for P. falciparum appearance was 12.6 days after the commencement of chloroquine treatment. Patients experiencing appearance of P. falciparum had significantly lower hematocrit, and greater initial P. vivax parasite counts. We use a mathematical model to explore the consequences of chloroquine treatment of such mixed infections. Both clinical results and features of the model suggest that such "hidden infections" may be quite common, and that the appearance of P. falciparum may be stimulated by treatment of P. vivax. PMID- 11485098 TI - Severe hepatic dysfunction associated with falciparum malaria. AB - We describe severe hepatic dysfunction associated with an attack of falciparum malaria in six Sri Lankan patients. Clinicians working in areas endemic for malaria should be made aware of this unusual complication. PMID- 11485099 TI - Tropical pulmonary eosinophilia and filariasis in Pakistan. AB - Filariasis is a major health problem in South Asia, particularly India, Sri Lanka and Bangladesh. Pakistan was presumed to be not affected. We report for the first time confirmed cases of tropical pulmonary eosinophilia (TPE) in indigenous patients as a result of infection with Wuchereria bancrofti. Following clinical examination, total leukocyte and eosinophil counts were recorded. Parasitological examinations included blood for microfilariae and stool and urine for eggs of intestinal parasites. Total immunoglobulin (Ig) E and specific antifilarial IgG were measured. Suspected cases of TPE were treated with diethylcarbamazine, 6 mg/kg for four weeks and were followed up to 2 and 4 weeks after treatment. Four persons fulfilled the criteria for TPE. Their response to treatment was marked with clinical improvement, reduction in eosinophil count and reduced titers of specific antifilarial antibodies. Two persons had W. bancrofti antigen in their sera confirmed by filariasis antigen detection test. Tropical pulmonary eosinophilia due to Wuchereria. bancrofti, although rare, is present in Pakistan. PMID- 11485100 TI - The application of ethanol-extracted Gloriosa superba for metaphase chromosome preparation in mosquitos. AB - The application of ethanol-extracted Gloriosa superba for metaphase chromosome preparation in adult and 4th larva Aedes aegypti revealed that 0.5-8% ethanol extracted Gl. superba solution could be used instead of 1% colchicine in Hanks' balanced salt solution. For adult mosquitos, the metaphase rates and average number of metaphase chromosomes per positive mosquito after intrathoracic inoculation with 1-2% ethanol-extracted Gl. superba solution were 100% and 11.8 (2-16) -12.6 (3-28) in females, and 80-90% and 16.5 (1-52) - 29.89 (1-72) in males, whereas the inoculation with 1% colchicine solution yielded 80% and 50% metaphase rates, and 18.25 (1-40) and 16.5 (2-53) average number of metaphase chromosomes per positive mosquito in females and males, respectively. For 4th stage larvae, the metaphase rates and average number of metaphase chromosomes per positive mosquito after incubation with 0.5-8% ethanol-extracted Gl. superba solution were 90-100% and 14.42 (1-65) - 64 (19-137), while incubation with 1% colchicine solution yield 100% metaphase rate and 10.9 (7-15) average number of metaphase chromosomes per positive mosquito. PMID- 11485101 TI - Preliminary studies of Anopheles mosquitos in eight provinces in Lao PDR. AB - Malaria vector surveys were carried out in 8 provinces in Lao PDR in 1999. The surveys were conducted in 4 provinces - Savannakhet, Champasak, Luang Perbang and Sayaboury in May and in another 4 provinces - Bolikhamsay, Sarvan, Sekong and Vientiane in December 1999. Bare leg collection were carried out indoors and outdoors from 6 pm to 5 am. All anopheline mosquitos were identified, dissected and the gut, gland and ovaries were examined. A total of 438 Anopheles mosquitos belonging to 19 species were obtained. Of these only 3 species were found to be infected with oocysts - An. maculatus, An. dirus and An. minimus. All these species were found biting both indoors and outdoors. An. aconitus was the predominant species obtained in the December collection but its vectorial status remains unknown. PMID- 11485102 TI - Inhibition of lymphocyte proliferative responses to Helicobacter pylori by plastic adherent cells. AB - A study was carried out on 49 H. pylori-positive and 11 H. pylori-negative patients to determine the reactivity of peripheral blood lymphocytes (PBL) to phytohemagglutinin (PHA) and acid glycine extract (AGE) of H. pylori, and to identify cells responsible for imunosuppression. Based on response to PHA stimulation, cell-mediated immunity of all patients were competent. In some patients, however, response to AGE of H. pylori was suppressed by plastic adherent cells. This study provided evidence of the presence of plastic adherent suppressor cells which suppressed PBL response to AGE of H. pylori but not to PHA suggesting that immunosuppression is antigen specific. There is also an indication that immunosuppression may be species-specific as PBL devoid of plastic adherent cells only responded to stimulation by AGE of H. pylori but not that to AGE of C. jejuni. PMID- 11485103 TI - Transition of drug susceptibilities of Vibrio cholerae O1 in Lao People's Democratic Republic. AB - The changes of drug susceptibilities of Vibrio cholerae O1 isolated during the past 7 years (1993-1999) in Lao PDR were investigated. The most noteworthy finding was the appearance of polymyxin B sensitive El Tor vibrios. Until 1996, the susceptibilities were almost as expected and cholera disappeared in 1997. When a cholera outbreak resurfaced in 1998, the susceptibilities of isolated V. cholerae O1 against tetracycline, sulfamethoxazol-trimethoprim, chloramphenicol and polymyxin B were quite different from those of previously isolated organisms. Minimum inhibitory concentrations (MICs) of tetracycline and chloramphenicol against the isolates in 1998 were about 16 times higher than those against the previous isolates, and the MICs of sulfamethoxazol-trimethoprim were about 256 times higher than those against the previous isolates, (trimethoprim 32 microg/ml: sulfamethoxazol 608 microg/ml). Eleven percent of the isolates (11/99) were as sensitive to polymyxin B as the classic cholera vibrios (MIC < 2 microg/ml). In 1999, the susceptibility pattern was almost the same as that in 1998 except for polymyxin B to which 58% of the isolates (21/36) became sensitive. PMID- 11485104 TI - Word use in the poetry of suicidal and nonsuicidal poets. AB - OBJECTIVE: The purpose of this study was to determine whether distinctive features of language could be discerned in the poems of poets who committed suicide and to test two suicide models by use of a text-analysis program. METHOD: Approximately 300 poems from the early, middle, and late periods of nine suicidal poets and nine nonsuicidal poets were compared by use of the computer text analysis program, Linguistic Inquiry and Word Count (LIWC). Language use within the poems was analyzed within the context of two suicide models. RESULTS: In line with a model of social integration, writings of suicidal poets contained more words pertaining to the individual self and fewer words pertaining to the collective than did those of nonsuicidal poets. In addition, the direction of effects for words pertaining to communication was consistent with the social integration model of suicide. CONCLUSIONS: The study found support for a model that suggests that suicidal individuals are detached from others and are preoccupied with self. Furthermore, the findings suggest that linguistic predictors of suicide can be discerned through text analysis. PMID- 11485105 TI - Religious coping, ethnicity, and ambulatory blood pressure. AB - OBJECTIVE: To investigate the relationship between religious coping, ethnicity, and ambulatory blood pressure (ABP) measured during daily life. METHODS: A 24 hour ABP was obtained from 155 men and women (78 African American and 77 white) on a typical workday. ABP was averaged over awake and sleep periods, and clinic BP was also assessed. Psychosocial measures of coping style, negative affect, social support, stress, and health behaviors were completed before ABP measurement. RESULTS: Multiple regression analyses, controlling for demographic variables, revealed a significant religious coping by ethnicity interaction for ABP (p < .01) and clinic BP (p < .05). Religious coping was not related to BP among whites. Among African Americans, however, higher levels of religious coping were associated with lower awake (p < .05) and sleep (p < .01) ABP. Social support satisfaction also was related to lower awake ABP among African Americans, but it did not mediate the relationship between religious coping and ABP. CONCLUSIONS: The results of this study extend previous findings by showing that, among African Americans, religious coping and BP are related during daily activities as well as in the clinic. Lower 24-hour BP load may be a pathway through which religiosity and cardiovascular health are related. PMID- 11485106 TI - Effects of psychological stress and psychiatric disorders on blood coagulation and fibrinolysis: a biobehavioral pathway to coronary artery disease? AB - OBJECTIVE: A hypercoagulable state before overt thrombosis resulting from an imbalance between the coagulation and fibrinolysis systems is related to cardiovascular disease progression and acute coronary syndromes. Psychological stressors and depressive and anxiety disorders also are associated with coronary artery disease. This review explores whether changes in blood coagulation, anticoagulant, and fibrinolytic activity may constitute psychobiological pathways that link psychological factors with coronary syndromes. METHODS: Literature on coagulation, anticoagulation, and fibrinolysis measures in conjunction with psychological factors (mental stress, psychosocial strain, and psychiatric disorders) was identified by MEDLINE search back to 1966 and through checking the bibliographies of these sources. Sixty-eight articles were critically reviewed. RESULTS: In healthy subjects, acute mental stress simultaneously activates coagulation (ie, fibrinogen or von Willebrand factor) and fibrinolysis (ie, tissue-type plasminogen activator) within a physiological range. In patients with atherosclerosis and impaired endothelial anticoagulant function, however, procoagulant responses to acute stressors may outweigh anticoagulant mechanisms and thereby promote a hypercoagulable state. Chronic psychosocial stressors (job strain or low socioeconomic status) are related to a hypercoagulable state reflected by increased procoagulant molecules (ie, fibrinogen or coagulation factor VII) and by reduced fibrinolytic capacity. There is also some evidence that points to hypercoagulability in depression. CONCLUSIONS: Different categories of psychological measures to varying extent are associated with characteristic patterns of coagulation and fibrinolysis activity. Associations between psychological factors and several coagulation and fibrinolysis variables related to atherosclerosis provide a plausible biobehavioral link to coronary artery disease. PMID- 11485107 TI - Sex, gender, and blood pressure: contributions to experimental pain report. AB - OBJECTIVE: The current study investigated whether the relationship between sex and experimental pain report was explained by systolic blood pressure (SBP) at rest or during pain task, by gender-role socialization as assessed by the Bem Sex Role Inventory, or both. The influence of gender-role socialization on pain report is often inferred but rarely studied. METHODS: Fifty female and 54 male healthy, young adults completed the Bem Sex Role Inventory and then underwent a cold pressor task. Blood pressure was assessed before and during pain testing. RESULTS: Univariate analyses indicated significant sex-related differences in pain threshold and pain tolerance. Baseline SBP was positively related to pain tolerance but did not explain sex differences, in accord with previous research. The Bem Sex Role Inventory demonstrated a relationship with pain, but did not explain sex differences. CONCLUSIONS: We suggest that context-specific measures of gender are needed to assess gender-related pain behaviors in specific situations. Results from the current study support our contention that gender is part of sex as commonly measured. Also, blood pressure does not appear to fully account for sex-related differences in pain. PMID- 11485108 TI - Depressed mood is a factor in glycemic control in type 1 diabetes. AB - OBJECTIVE: The diabetes literature contains conflicting evidence on the relationship between depression and glycemic control. This may be due, in part, to the fact that past studies failed to distinguish between patients with type 1 and type 2 diabetes. Because these are actually completely different diseases that are often treated differently and consequently make different demands on patients, the relationship between glycemic control and depressed mood in type 1 and type 2 diabetes was examined separately. METHODS: The relationship between Beck Depression Inventory (BDI) scores and HbA1c, as an index of long-term glycemic control, was measured in samples of 30 patients with type 1 and 34 patients with type 2 diabetes. RESULTS: Groups of patients with type 1 and type 2 diabetes did not differ in mean BDI score or HbA1c level. Correlation analysis revealed a significant positive relationship between BDI scores and HbA1c in the type 1 group (r = .44, p < .02) but not in the type 2 group (r = -0.06, p > .05). This relationship was evident throughout the entire range of BDI scores and was not restricted to scores indicative of clinical depression. Patients with type 1 diabetes who had higher HbA1c and BDI scores reported a lower frequency of home blood glucose monitoring. CONCLUSIONS: Variations in depressive mood, below the level of clinical depression, are associated with meaningful differences in glycemic control in type 1 but not type 2 diabetes. Preliminary data analysis suggests that this effect may be mediated, at least in part, by decreased self care behaviors in patients with more depressed mood. PMID- 11485109 TI - Attachment and psychosomatic medicine: developmental contributions to stress and disease. AB - OBJECTIVE: The object of this study was to evaluate the evidence linking attachment insecurity to illness. Attachment theory describes lifelong patterns of response to threat that are learned in the interaction between an infant and his or her primary caregiver. Despite its biopsychosocial domain, attachment theory has only recently been applied to psychosomatic medicine. METHOD: MEDLINE and PsychInfo databases were searched from 1966 to 2000 for English language papers with key words "attachment" and "object relations." Papers and their cited references were reviewed if they were directly related to physical illness, symptoms, or physiology. A hypothetical causal model was developed. RESULTS: Direct and indirect evidence from survey studies supports an association between attachment insecurity and disease. Animal studies and human experiments suggest that attachment contributes to individual differences in physiological stress response. There is also less robust support for insecure attachment leading to symptom reporting and to more frequent health risk behaviors, especially substance use and treatment nonadherence. Evidence supports the prediction from attachment theory that the benefits of social support derive more from attachment relationships than nonattachment relationships. CONCLUSIONS: Although the available data are suggestive rather than conclusive, the data can be organized into a model that describe attachment insecurity leading to disease risk through three mechanisms. These are increased susceptibility to stress, increased use of external regulators of affect, and altered help-seeking behavior. This model warrants further prospective investigation. PMID- 11485110 TI - Effects of sleep on endotoxin-induced host responses in healthy men. AB - OBJECTIVE: To examine whether increased sleep during viral or bacterial infections supports host defense mechanisms. METHODS: To test this assumption in humans, healthy male subjects were assigned either to sleep from 2300 to 0700 hours (n = 10) or to stay awake through the night (n = 10). In the sleeping subjects Salmonella abortus equi endotoxin (0.4 ng/kg) or placebo were intravenously injected in balanced order during the first SWS episode. The age matched, sleep-deprived subjects were injected at the same time point. RESULTS: As expected, endotoxin significantly increased rectal temperature, the plasma levels of cortisol, tumor necrosis factor-alpha (TNF-alpha), the soluble TNF receptors p55 and p75, Interleukin (IL)-6, the IL-1 receptor antagonist (RA), leukocyte, and granulocyte counts in both sleeping and sleep-deprived subjects, whereas lymphocyte and monocyte counts were transiently reduced. Time courses of endotoxin-induced host responses did not differ between the sleep and sleep deprivation groups. Endotoxin did not affect the amount of nocturnal wakefulness, nonrapid-eye-movement (NREM) sleep, or rapid-eye-movement (REM) sleep across the total night compared with placebo, but significantly increased electroencephalogram-arousals (EEG-arousals) in stage 2 and decreased arousals in SWS. In addition, the amount of SWS, spectral EEG-delta and -theta power was increased at the beginning and at the end of the sleep period, respectively, when the degree of immune activation was relatively low. CONCLUSION: The present results support the notion that short-term sleep deprivation is unlikely to harm the immune system as far as unspecific acute responses are concerned. The effects of endotoxin on sleep in this case support prior observations that in humans, enhanced SWS and intensified NREM sleep occur when host defense activation is subtle. PMID- 11485111 TI - Neuropsychological effects of one-week continuous positive airway pressure treatment in patients with obstructive sleep apnea: a placebo-controlled study. AB - OBJECTIVE: To determine whether 1-week continuous positive airway pressure (CPAP) treatment, compared with placebo CPAP, improves cognitive functioning in patients with obstructive sleep apnea (OSA). METHODS: 36 OSA patients (aged 32-60 years, respiratory disturbance index [RDI] > 15) were monitored 2 nights with polysomnography, then randomized for 1-week treatment to CPAP or placebo (CPAP at 2 cm H2O with holes in mask). Participants completed Wechsler Adult Intelligence Scale-Revised Digit Symbol and Digit Span, Trailmaking A/B, Digit Vigilance, Stroop Color-Word, Digit Ordering, and Word Fluency tests pre- and posttreatment. These produced 22 scores per participant, which were analyzed by use of repeated measures analysis of variance (ANOVA) and a rank-sum test. RESULTS: In ANOVA, only 1 of the 22 scores showed significant changes specific to CPAP treatment, a number that could be expected by chance alone: Digit Vigilance-Time (p = .035). The CPAP group improved their time (from 7.5 to 6.9 minutes. p = .013). The rank sum test revealed that the CPAP group had significantly better overall cognitive functioning posttreatment than the placebo group (mean ranks of 17.8 vs. 20.2, respectively; p = .022). CONCLUSIONS: Although results suggest overall cognitive improvement due to CPAP, no beneficial effects in any specific cognitive domain were found. Future studies of neuropsychological effects of CPAP treatment should include a placebo CPAP control group. Placebo studies that use longer-term treatment might demonstrate additional effects. It is also possible that, even at 2 cm H2O, CPAP conveys some beneficial neuropsychological effects. PMID- 11485112 TI - A meta-analytic examination of basal cardiovascular activity in posttraumatic stress disorder. AB - OBJECTIVE: The objective of this meta-analytic study was to determine whether individuals with posttraumatic stress disorder (PTSD) have higher levels of basal cardiovascular activity relative to comparable groups of individuals without PTSD. METHODS: Meta-analytic data methods were applied to 34 studies that gathered indicators of basal cardiovascular activity including: heart rate (HR), systolic blood pressure, and diastolic blood pressure on subjects diagnosed with PTSD and two types of comparison groups. In total, cardiovascular measures were analyzed for 2670 subjects across all studies. RESULTS: Results indicate that individuals with a current PTSD diagnosis have higher resting HR relative to both trauma-exposed individuals without a PTSD diagnosis and non-trauma-exposed individuals. The results also suggest that PTSD is associated with elevations in blood pressure; however, the effect sizes were smaller in magnitude than those obtained for heart rate. A subset analysis revealed that the effect sizes for comparisons on basal HR were greatest in studies with the most chronic PTSD samples. CONCLUSION: The meta-analysis supports previous qualitative reviews, finding a positive association between PTSD and basal cardiovascular activity. The discussion addresses possible mechanisms of action and the health-related implications of these findings. PMID- 11485113 TI - Somatization symptoms and hypochondriacal features in the general population. AB - OBJECTIVE: The principal goal of this study is to examine the base rates of somatoform symptoms and of hypochondriacal features in the general population. METHODS: A representative sample of 2050 persons in Germany was examined by use of screening for somatoform symptoms and the Whiteley Index. RESULTS: The most frequent somatoform symptoms were back pain, joint pain, pain in extremities, and headache, as well as abdominal symptoms (bloating or intolerance of several foods) and cardiovascular symptoms (palpitation). People reported a mean of two somatization symptoms of DSM-IV somatization disorder (SD) during the prior 2 years. Strong age and medium gender effects were found for most somatoform symptoms, as well as for composite indices. However, the sex ratio suggested in DSM-IV for SD seems to be an overestimation. Hypochondriacal features showed only small sex differences but, again, pronounced age effects. In contrast to low rates for SD, the base rates for somatization and hypochondriacal features were high and represented the health care relevance of subthreshold syndromes. CONCLUSION: We present base rates of hypochondriacal and somatization features that may be important facets in the development of classification criteria and in the interpretation of health care expenditure. PMID- 11485114 TI - Depressive symptoms, menopausal status, and climacteric symptoms in women at midlife. AB - OBJECTIVE: Previous studies have found increased rates of depression in women aged 45 to 54 years, but the factors that influence these rates are not understood. It was assessed whether higher rates of depressive symptoms were associated with menopausal status, climacteric symptoms, and use of hormone replacement therapy. DESIGN: Cross-sectional survey. SETTING: Community sample. METHODS: Data are from 581 women ages 45 to 54 years who were interviewed by telephone between October 1998 and February 1999. MEASURES: Depression was measured with the abbreviated CES-D, a depressive symptoms screening measure. Women's reported perception of menopausal stage, frequency of periods in the preceding 12 months, and history of oophorectomy were used to classify their menopausal status into four categories: (1) no indication of menopause; (2) close to menopause; (3) had begun menopause; and (4) had completed menopause. RESULTS: There were 168 women (28.9%) who reported a high level (> or = 10) of depressive symptoms when the abbreviated CES-D was used. In a logistic-regression analysis, significant factors associated with increased depressive symptoms included physical inactivity, inadequate income, use of estrogen/progesterone combination, and presence of climacteric symptoms (trouble sleeping, mood swings, or memory problems). Menopausal status was not associated with depressive symptoms. CONCLUSIONS: In this sample of women age 45 to 54 years, climacteric symptoms but not menopausal status were associated with higher rates of depressive symptoms. PMID- 11485115 TI - Social support and health behavior in hostile black and white men and women in CARDIA. Coronary Artery Risk Development in Young Adults. AB - OBJECTIVE: These cross-sectional analyses of the Coronary Artery Risk Development in Young Adults (CARDIA) data were stimulated by previous CARDIA analyses that showed an adverse association between hostility and several health behaviors: physical activity, cigarette smoking, alcohol consumption, and caloric intake, in both black and white men and women, such that the higher the hostility, the worse the health behavior profile. The current study investigated whether high social support was associated with better health behavior than low social support in individuals with high hostility scores. METHODS: The subjects were 5115 healthy black and white men and women ranging in age from 18 to 30 years. The hypothesis was that the association between hostility and certain adverse health behaviors would be diminished in the presence of high social support. Race-gender specific median cutpoints of the Cook-Medley Hostility scale and an index of social support defined levels of high and low hostility and social support. RESULTS: After controlling for age and body mass index (BMI), support was positively associated with more exercise in all groups except black women, but when coupled with high hostility, this positive association between support and exercise remained only in men. White women with high support were less often smokers but this association did not hold when examined only in the high-hostile group. Black men and white women with high support in the presence of high hostility consumed more alcohol, but the amount was moderate. CONCLUSIONS: We conclude that social support in the presence of high hostility only sometimes reduces the association of hostility to adverse health behaviors and that these effects are complex. Additional research investigating types of social support on health behavior in different race-gender groups is advocated. PMID- 11485116 TI - Association of depression and diabetes complications: a meta-analysis. AB - OBJECTIVE: The objective of this study was to examine the strength and consistency of the relationship between depression and diabetes complications in studies of type 1 and type 2 adult patients with diabetes. METHOD: MEDLINE and PsycINFO databases were searched for articles examining depression and diabetes complications in type 1 and type 2 diabetes samples published between 1975 and 1999. Meta-analytic procedures were used. Studies were reviewed for diabetes type, sample size, statistical tests, and measures of diabetes complications and depression. Significance values, weighted effect sizes r, 95% confidence intervals (CI), and tests of homogeneity of variance were calculated for the overall sample (k = 27) and for subsets of interest. RESULTS: A total of 27 studies (total combined N = 5374) met the inclusion criteria. A significant association was found between depression and complications of diabetes (p < .00001, z = 5.94). A moderate and significant weighted effect size (r = 0.25; 95% CI: 0.22-0.28) was calculated for all studies reporting sufficient data (k = 22). Depression was significantly associated with a variety of diabetes complications (diabetic retinopathy, nephropathy, neuropathy, macrovascular complications, and sexual dysfunction). Effect sizes were in the small to moderate range (r = 0.17 to 0.32). CONCLUSIONS: These findings demonstrate a significant and consistent association of diabetes complications and depressive symptoms. Prospective, longitudinal studies are needed to identify the pathways that mediate this association. PMID- 11485117 TI - Magnetic resonance spectroscopic and relaxometric determination of bone marrow changes in anorexia nervosa. AB - OBJECTIVE: The objective of this study was to assess and quantify bone marrow changes in patients with anorexia nervosa using 1H magnetic resonance spectroscopy and relaxometry. METHODS: The bone marrow fat fraction and the longitudinal and transverse relaxation times (T1 and T2, respectively) of water were measured in the lumbar and femoral marrow of 20 patients with anorexia nervosa and 19 healthy control subjects. RESULTS: Patients with anorexia nervosa showed significant hyperhydration and reduction of the fat fraction in their bone marrow, predominantly in the proximal femur. These changes were associated with hematological abnormalities. In a retest of seven patients after psychotherapy and gain of weight, the pathological changes in marrow proved to be largely reversible in correlation with the increase in body mass index. CONCLUSIONS: Fat depletion and excess of tissue water in the bone marrow in anorexia nervosa can be quantified by 1H magnetic resonance spectroscopy and relaxometry. The distribution of the pathological changes in the lumbar and femoral marrow follows the pattern of normal bone marrow conversion from hematopoietic to cellular during childhood. PMID- 11485118 TI - Slow recovery from voluntary hyperventilation in panic disorder. AB - OBJECTIVE: Because hyperventilation has figured prominently in theories of panic disorder (PD) but not of social phobia (SP), we compared predictions regarding diagnosis-specific differences in psychological and physiological measures before, during, and after voluntary hyperventilation. METHOD: Physiological responses were recorded in 14 patients with PD, 24 patients with SP, and 24 controls during six cycles of 1-minute of fast breathing alternating with 1 minute of recovery, followed by 3 minutes of fast breathing and 10 minutes of recovery. Speed of fast breathing was paced by a tone modulated at 18 cycles/minute, and depth by feedback aimed at achieving an end-tidal pCO2 of 20 mm Hg. These values were reached equally by all groups. RESULTS: During fast breathing, PD and SP patients reported more anxiety than controls, and their feelings of dyspnea and suffocation increased more from baseline. Skin conductance declined more slowly in PD over the six 1-minute fast breathing periods. At the end of the final 10-minute recovery, PD patients reported more awareness of breathing, dyspnea, and fear of being short of breath, and their pCO2s, heart rates, and skin conductance levels had returned less toward normal levels than in other groups. Their lower pCO2s were associated with a higher frequency of sigh breaths. CONCLUSIONS: PD and SP patients report more distress than controls to equal amounts of hypocapnia, but PD differ from SP patients and controls in having slower symptomatic and physiological recovery. This finding was not specifically predicted by hyperventilation, cognitive-behavioral, or suffocation alarm theories of PD. PMID- 11485119 TI - Vagal rebound and recovery from psychological stress. AB - OBJECTIVE: To characterize cardiovascular recovery and examine the possible relationship of vagal activity and reflexes to risk for heart disease. METHODS: Subjects performed cold pressor and mental arithmetic tasks. Heart rate, heart period variability, and pre-ejection period were obtained for 1 minute before, during, and after each task (Experiment 1). In the second experiment, subjects performed a Stroop color-word task and a mental arithmetic task. Heart rate, heart period variability, blood pressure, and baroreflex sensitivity were obtained during the 5-minute baseline, task, and recovery periods (Experiment 2). RESULTS: In Experiment 1, heart rate during recovery was lower than baseline despite continued pre-ejection period shortening, whereas recovery heart period variability was higher than baseline. In Experiment 2, blood pressure increased throughout the session. However, recovery heart rate after mental arithmetic was lower than baseline heart rate, and heart period variability was higher during both recovery periods than during baseline. Vagal rebound, a sharp increase in variability in the first minute of recovery, was reduced in men in Experiment 1 and in individuals with a family history of cardiovascular disease in Experiment 2 and was associated with degree of change in baroreflex sensitivity between task and rest. CONCLUSIONS: Cardiovascular recovery from stress is associated with increased vagal modulation despite residual sympathetic activation. Vagal rebound may be involved in mechanisms resetting the baroreflex sensitivity at the onset and offset of stress. Diminished vagal rebound during recovery from stress is associated with standard risk factors for cardiovascular disease. The results support an association between attenuated vagal reflexes and risk for cardiovascular disease. PMID- 11485120 TI - Intrusion, avoidance, and psychological distress among individuals with cancer. AB - OBJECTIVE: The goal of the study was to examine the utility of Creamer's cognitive processing theory of trauma in a sample of individuals undergoing treatment for cancer. This theory proposes that avoidance is a maladaptive strategy of dealing with intrusive thoughts about a traumatic experience and suggests that avoidance mediates the relation between intrusive thoughts and later psychological distress. The role of disease-related factors, specifically changes in physical impairment and disease stage, was also examined. METHODS: Patients (N = 189) undergoing treatment for cancer completed questionnaires at three time points, spaced 3 months apart. Intrusive thoughts, functional impairment, and psychological distress were assessed at Time 1, avoidance and functional impairment at Time 2, and psychological distress was assessed again at Time 3. The fit of the model was tested separately for patients with early-stage (stages 1 and 2) and late-stage (stages 3 and 4) disease. RESULTS: The mediational role for avoidance was supported among patients with advanced stages of cancer but not for patients with early-stage disease. Results were inconsistent with predictions about the role of physical impairment. Among individuals with late-stage cancer, changes in functional impairment were not predictive of greater avoidance, and impairment had a significant but weak effect on the change in distress. Among patients with early-stage cancer, a deterioration in physical impairment was associated with increases in avoidance, and deterioration in physical impairment increased distress. CONCLUSIONS: The results of this study were partially consistent with Creamer's cognitive processing theory. A moderating effect was found for disease stage on associations between intrusions, avoidance, physical impairment, and distress. PMID- 11485121 TI - Development of the stress response inventory and its application in clinical practice. AB - OBJECTIVE: The purpose of this study was to develop the Stress Response Inventory (SRI), which includes emotional, somatic, cognitive, and behavioral stress responses, and then to use the scale in clinical practice. METHODS: First, a preliminary survey was conducted using 109 healthy adults to obtain 75 response items. Second, the preliminary questionnaire was completed by 215 healthy subjects. Third, stress responses were compared among 242 patients (71 with anxiety disorder, 73 with depressive disorder, 47 with somatoform disorder, and 51 with psychosomatic disorder) and the 215 healthy subjects. RESULTS: Factor analysis yielded seven subscales: tension, aggression, somatization, anger, depression, fatigue, and frustration. Reliability was computed by administering the SRI to 62 healthy subjects during a two-week interval. Test-retest reliability for the seven subscale scores and the total score was high, ranging between 0.69 and 0.96. Internal consistency was computed, and Cronbach's alpha for the seven subscales ranged between 0.76-0.91 and 0.97 for the total score. Convergent validity was computed by correlating the seven subscales and the total score of the SRI with the total score of the Global Assessment of Recent Stress (GARS) scale, the Perceived Stress Questionnaire (PSQ), and the subscale scores of the Symptom Checklist-90-Revised (SCL-90-R). The correlations were all at significant levels. The sensitivity of the SRI was 0.57, specificity 0.74, and the predictive value positive (PVP) was 0.71. The patient group also scored significantly higher on the six subscale scores and the total score than the control group, with the exception being the aggression subscale. The depressive disorder group was highest in total scores on the SRI among the four patient groups, and showed significantly higher total scores than the anxiety disorder and psychosomatic disorder groups. In total scores on the SRI, female subjects scored significantly higher than males. CONCLUSIONS: These results indicate that the SRI is highly reliable and valid, and that it can be utilized as an effective measure of stress for research in stress-related fields. The depressive disorder group showed more prominent stress responses than the anxiety and psychosomatic disorder groups. PMID- 11485122 TI - Validation and utility of the patient health questionnaire in diagnosing mental disorders in 1003 general hospital Spanish inpatients. AB - OBJECTIVE: To determine whether the Spanish version of the patient health questionnaire (PHQ) has validity and utility for diagnosing mental disorders in general hospital inpatients. METHODS: Participants in the study were 1003 general hospital inpatients, randomly selected from all admissions over an 18-month period. All of them completed the PHQ, the Beck Depression Inventory (BDI), and measures of functional status, disability days, and health care use, including length of hospital stay. They also had a structured interview with a mental health professional. RESULTS: A total of 416 (42%) of the 1003 general hospital inpatients had a PHQ diagnosis. There was good agreement between PHQ diagnoses and those of an independent mental health professional (for the diagnosis of any PHQ disorder, kappa = 0.74; overall accuracy, 88%; sensitivity, 87%; specificity, 88%), similar to the original English version of the PHQ in primary care patients. Patients with PHQ diagnoses had more functional impairment, disability days, and health care use than did patients without PHQ diagnoses (group main effects for functional status measures and disability days, p < .001; group main effects for health care use, p < .01). The group main effect for hospital length of stay was not significant. An index of depression symptom severity calculated from the PHQ correlated significantly both with the number of depressive symptoms detected at interview and the total BDI score. PHQ administration was well accepted by patients. CONCLUSIONS: The Spanish version of the PHQ has diagnostic validity in general hospital inpatients comparable to the original English version in primary care. PMID- 11485123 TI - Depression and reactivity to stress in older women with rheumatoid arthritis and osteoarthritis. AB - OBJECTIVE: The purpose of this study was to examine the role of depressive symptoms in reactivity to stress and pain in older women with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: Participants were 188 older women with RA (N = 87) and OA (N = 101). They were initially assessed for depressive symptoms and interviewed weekly for 12 to 20 weeks regarding interpersonal stress, arthritis pain, and negative affect. RESULTS: Hierarchical linear modeling (HLM) revealed that depressive symptoms were related to weekly elevations in arthritis pain, negative events, perceived stress, and negative affect for RA respondents and elevations in arthritis pain and negative affect for OA respondents. HLM analyses also indicated that depressive symptoms were related to increased reactivity to perceived stress and arthritis pain in people with RA, but not those with OA. CONCLUSIONS: Depression may be related to elevations in pain for people with RA and OA and to elevations in stress and increased reactivity to stress and pain for those with RA. PMID- 11485124 TI - Sulfasalazine decreases acute gastrointestinal complications due to pelvic radiotherapy. AB - BACKGROUND: Radiation-induced gastrointestinal toxicity is a significant concern for patients who are treated with this modality for pelvic malignancies. Eicosanoids and free radicals are thought to be among the reasons for this effect. Sulfasalazine is an inhibitor of their synthesis in the mucosa. OBJECTlVE: To determine whether sulfasalazine can reduce the radiation-induced acute gastrointestinal complications. METHODS: In this prospective, double-blind study, 31 patients receiving pelvic radiotherapy were randomized to receive two sulfasalazine 500-mg tablets twice daily or placebo, administered orally from the first day of irradiation. Patients were evaluated weekly, and gastrointestinal toxicities were graded according to the Late Effect of Normal Tissue-Subjective Objective Management Analytic (LENT-SOMA) toxicity table during pelvic radiotherapy. On the last day of week 5, the subjects were graded endoscopically, and biopsies taken from the rectum were classified histopathologically. RESULTS: Groups did not differ in age, gender, tumor site, or irradiation procedure. During radiotherapy, grade 2 or higher gastrointestinal toxicity occurred in 20% (3/15) and 63% (10/16) of the sulfasalazine and placebo groups, respectively. This difference was significant (p = 0.017). No statistically significant differences were found in endoscopic and histopathologic evaluations. CONCLUSIONS: Sulfasalazine is effective in decreasing clinically acute gastrointestinal toxicities. Long-term follow-up with the subjects will help to determine the net effect of sulfasalazine on the radiation-induced gastrointestinal injuries. PMID- 11485125 TI - Use of antihypertensive and antithrombotic medications after stroke in community based care. AB - BACKGROUND: Secondary stroke prevention strategies include pharmacologic approaches to control hypertension and reduce thromboembolic risk. OBJECTIVE: To describe antithrombotic and antihypertensive medication use, and rates of blood pressure control in the Kansas City Stroke Study, a prospective stroke cohort receiving community-based care after primarily mild and moderate stroke. METHODS: Participants from 12 area hospitals provided information about medication use prior to stroke. Study personnel measured blood pressures at enrollment and at one, three, and six months, and collected medication data at six months during in home assessment. RESULTS: Complete data at six months were available for 355 subjects with ischemic stroke, among whom 13% had atrial fibrillation and 67% had prior hypertension. Prior to stroke, only 45% of the patients were receiving any antithrombotic (anticoagulant and/or antiplatelet) therapy; this figure rose to 77% at six months. Antithrombotic treatment rates among those with atrial fibrillation were 59% before stroke and 83% at six months, including warfarin in 64%. Approximately 70% of subjects had controlled blood pressures one, three, and six months after stroke, defined as systolic blood pressure < or = 140 mm Hg and diastolic blood pressure < or = 90 mm Hg. Use of multiple antihypertensive agents was common; calcium-channel blockers and angiotensin-converting enzyme inhibitors were used most frequently. However, 19% of subjects with uncontrolled blood pressure were untreated at six months. CONCLUSIONS: Although room for improvement remains, these data suggest improved rates of antithrombotic and antihypertensive medication use after stroke in community-based care in a midwestern metropolitan community, compared with previous reports. PMID- 11485126 TI - Blood pressure monitoring with home monitors versus mercury sphygmomanometer. AB - OBJECTIVE: To determine the accuracy of three automatic monitors (arm, wrist, finger) for blood pressure measurement manufactured by Omron compared with a standard mercury sphygmomanometer. PRIMRY END POINT: Difference in the mean blood pressure readings from each monitor; the secondary end point was difference in pulse readings. DESIGN: A single-visit, crossover trial tested each device twice on the left arm of each participant; the average of the two readings was recorded. The pulse readings from each monitor were also recorded. ANOVA was used to compare mean blood pressure readings and pulse readings from each device. RESULTS: A total of 55 persons (mean age 53 y; 36 women) met inclusion criteria and completed the study. The mean systolic and diastolic readings obtained from the electronic arm unit were comparable to the mercury readings (124.4/78.02 vs. 129.45/77.87 mm Hg, respectively; p > 0.05 for both readings). The mean results obtained from the wrist and finger monitors differed significantly from those of the mercury readings (145.44/89.58 and 113.94/69.07 mm Hg, respectively; p < 0.05 for both monitors compared with control). No difference was measured in the mean pulse readings between the comparisons (p = 0.72). The absolute difference in systolic and diastolic blood pressure readings from control varied the least wih the arm monitor. CONCLUSIONS: Compared with the mercury sphygmomanometer, the arm monitor was the most accurate in measuring blood pressure. The wrist and finger monitors resulted in statistically significant mean systolic and diastolic differences compared with the mercury sphygmomanometer. PMID- 11485127 TI - Use of infliximab in pediatric patients with inflammatory bowel disease. AB - BACKGROUND: The concentration of tumor necrosis factor, a proinflammatory cytokine, is increased in the gastrointestinal mucosa of patents with active Crohn's disease (CD) and ulcerative colitis (UC). Neutralization of tumor necrosis factor decreases the mucosal inflammatory response of adults with CD. Little information is available on the use of monoclonal antibody to tumor necrosis factor (infliximab) in children and adolescents with CD or UC. OBJECTIVE: To evaluate the clinical response and side effects of patients to infliximab. METHODS: A retrospective review of data regarding 18 pediatric and adolescent patients with active CD (n = 15) and UC (n = 3) poorly controlled with conventional therapy. All patients received one to six intravenous infusions of infliximab 5 mg/kg, while receiving their usual medications. RESULTS: All patients experienced clinical improvement, including decrease in the frequency of stooling and resolution of extraintestinal symptoms such as arthropathy, malaise, and skin manifestations after treatment with infliximab. All but one patient had a documented decrease in the erythrocyte sedimentation rate. Prednisone dosage was tapered in all but two patients, and discontinued in seven patients. Intravenous infusion of infliximab was well tolerated. One patient developed a rash several days after the infusion. A patient who received six infliximab infusions developed recurrent Staphylococcus aureus infections, as well as septic arthritis and chronic osteomyelitis during the follow-up period, raising the issue of the long-term safety of infliximab. CONCLUSIONS: Treatment of our patients with refractory CD and UC with infliximab was associated with remarkable clinical improvement. Although the drug may have an important role in their management, further assessment of long-term safety and efficacy is needed. PMID- 11485128 TI - Upper gastroduodenal ulceration in arthritis patients treated with celecoxib. AB - OBJECTIVE: To evaluate the comparative incidence of endoscopic gastroduodenal ulcers in patients with rheumatoid arthritis or osteoarthritis treated with celecoxib. DESIGN: Quantitative systematic review of randomized controlled trials. SUBJECTS: Patients (n = 4632) with rheumatoid arthritis or osteoarthritis reported in five trials. MAIN OUTCOME MEASURES: Rate ratios, rate differences, and the number needed to harm were calculated for the incidence of endoscopically documented gastroduodenal ulcers. RESULTS: Pooled rate ratios (RRs) relative to placebo for endoscopic ulcers at 12 weeks were 1.96 (95% CI 0.85 to 4.55) for celecoxib 100 mg twice daily and 2.35 (95% CI 1.02 to 5.38) for celecoxib 200 mg twice daily. There was no significant difference in gastroduodenal ulcers at 12 weeks between celecoxib 200 mg twice daily and celecoxib 100 mg twice daily; the corresponding pooled RR was 1.21 (95% CI 0.62 to 2.38). In contrast, celecoxib 200 mg twice daily was associated with a significantly lower rate of gastroduodenal ulcers than was naproxen 500 mg twice daily at 12 weeks (RR 0.24; 95% CI 0.17 to 0.33). On average, for every seven patients treated with naproxen, one more had an endoscopic ulcer than if they were treated with celecoxib. Celecoxib 200 mg twice daily also had a significantly lower risk of endoscopic ulcers than did either modified-release diclofenac 75 mg twice daily at 24 weeks (RR 0.24; 95% CI 0.11 to 0.52) or ibuprofen 800 mg three times daily at 12 weeks (RR 0.30; 95% CI 0.20 to 0.46). CONCLUSIONS: Endoscopic studies have shown that celecoxib, at a wide range of doses, is associated with a lower incidence of gastroduodenal ulcers than are diclofenac, ibuprofen, or naproxen. The incidence rates of gastroduodenal ulcers associated with celecoxib were similar, although not equivalent, to placebo. Head-to-head comparisons suggest that, at the wide range of doses studied (100-800 mg/d), there are no dose-related increases in endoscopic gastroduodenal ulcers with celecoxib. The results of longer term comparative trials of celecoxib based on clinical outcomes are needed to determine celecoxib's ultimate risk-benefit profile. PMID- 11485129 TI - Pharmacotherapy consultation on polypharmacy patients in ambulatory care. AB - OBJECTIVE: To investigate actual cost and adverse effect outcomes associated with a phamacotherapy consultation in ambulatory care patients receiving polypharmacy. METHODS: Patients receiving five or more chronic medications were randomized to receive pharmacotherapy consultation or usual medical care. Outcomes measured were changes in drug costs, medical costs, and drug-related symptoms six months after the consultation. Data were analyzed with unpaired Student's t-test for continuous data. Chi2 Analysis was used for categorical data. Patients and physicians were surveyed about their perceptions of the consultations after the study period. RESULTS: Drug and medical costs did not differ before and after the consultation. More patients in the consultation group had adverse symptom scores improve by two or more points, and fewer had symptom scores worsen by two or more points than in the control group. Seventy percent of patients and 76% of physicians believed that the consult was beneficial. CONCLUSIONS: Polypharmacy patients are the most likely to have drug-related problems and require intervention. Of all the interventions performed in this study, 73% of the original problems were recognized only through a patient interview, suggesting that an interpersonal relationship remains critical to the provision of pharmaceutical care. Although patients and physicians see intuitive value in pharmaceutical care, pharmacists need to exert more energy in the direction of marketing the profession. Finally, there are numerous difficulties in measuring the benefits of these interventions, possibly making broad-based interventions in complicated patients too difficult to assess accurately. Future studies should focus on patients with limited, specific problems or on interventions with narrow goals. PMID- 11485130 TI - Pharmacists' attitudes toward diabetes and their involvement in diabetes education. AB - OBJECTIVE: To determine the relationship between pharmacists' attitudes toward diabetes and their involvement in diabetes patient education in the community setting. METHODS: Registered pharmacists in Arizona were mailed surveys regarding their attitudes toward diabetes and their involvement in diabetes patient education. Attitudes were measured using the Diabetes Attitude Scale (DAS); the types of educational skills evaluated were based on those recommended by the American Diabetes Association's Standards of Medical Care. RESULTS: Pharmacists' attitudes were significantly positive toward the need for special training for diabetes care, the importance of tight glycemic control, the team approach to care, and the preference for diabetes education in an outpatient setting (p < 0.001). The majority of the time, pharmacists provided basic patient education (52%) rather than intermediate or advanced patient education (26% and 27%, respectively). There was a negative correlation between the attitude that diabetes is a difficult disease to treat and pharmacists' involvement in diabetes patient education (p < 0.05). This indicates that, although pharmacists believe that diabetes is a treatable disease, they infrequently provide diabetes patient education. CONCLUSIONS: Overall, pharmacists had positive attitudes toward diabetes. These attitudes did not correlate with the degree of their involvement in diabetes patent education. More diabetes patient education through community pharmacists is needed. PMID- 11485131 TI - Cost-effectiveness results from the US Carvedilol Heart Failure Trials Program. AB - OBJECTIVE: To evaluate the cost-effectiveness of carvedilol, a beta-blocker that is approved for use in the US for the treatment of heart failure, based on data from Phase III clinical trials. METHODS: We conducted an economic evaluation alongside the US Carvedilol Heart Failure Trials Program, which consisted of four concurrent, randomized, double-blind, placebo-controlled clinical trials; the mean duration of follow-up across these four trials was 6.5 months (the program was terminated prematurely based on a finding of a 65% mortality benefit). Using data from these trials, we examined the cost-effectiveness of carvedilol in terms of the estimated cost per death averted among patients randomized to such therapy versus those receiving placebo. Attention was focused on the cost of carvediol therapy plus the cost of cardiovascular-related inpatient care. Costs of care were estimated by combining infomation on healthcare utilization from the clinical trials with secondary sources of cost data. RESULTS: Patients randomized to receive carvedilol had lower mean +/- SD estimated costs of cardiovascular related inpatient care over 6.5 months compared with those receiving placebo ($1912 +/- $7595 vs. $4463 +/- $20,565, respectively). As mortality alsowas lower among carvedilol patients, the estimated cost per death averted was negative. The probability that carvedilol would both increase survival and decrease costs of cardiovascular-related care over a 6.5-month period was estimated to be 0.98. CONCLUSIONS: Data from the US Carvedilol Heart Failure Trials Program indicate that carvedilol reduces mortality in patients with heart failure; our study suggests that it also may be cost-saving over a period of approximately six months. PMID- 11485132 TI - Impact of a limited fluoroquinolone reimbursement policy on antimicrobial prescription claims. AB - OBJECTIVE: To examine the influence on administrative pharmacy claims of a policy that limited the reimbursement of the fluoroquinolones and other antimicrobials in the senior population within Nova Scotia, Canada. METHODS: The administrative claims database of the Nova Scotia Seniors' Pharmacare Program was used to identify all prescription claims for orally administered antibiotics and urinary antiinfectives. The number of beneficiaries receiving antimicrobials and the number, duration, and cost of prescriptions for antimicrobials were measured monthly. Descriptive time-series plots were used to compare antimicrobial use for two 12-month periods before the institution of the policy (December 1, 1994 November 30, 1995, and December 1, 1995-November 30, 1996) and the 12 months after the policy took effect (January 1, 1997-December 31, 1997). RESULTS: Following the implementation of the fluoroquinolone reimbursement policy, the number of patients using antimicrobials decreased by 2.2% and the number of prescriptions for antimicrobials decreased by 3.4%. Fluoroquinolone prescriptions decreased by 80.2%; prescriptions for sulfonamides and trimethoprim increased by 34.9%, cephalosporins by 17.0%, and macrolides and lincosamides by 16.5%. The only prescription duration to change was the fluoroquinolones, which increased by 25%. The average cost per antimicrobial user/year decreased from $35.24 during prepolicy period 2 to $27.51 during the postpolicy period. CONCLUSIONS: Prescription claims for fluoroquinolones in seniors decreased following the introduction of the policy. Total antimicrobial use also decreased, although this may be related to other factors. The effect of this policy change on patient outcomes requires further study. PMID- 11485133 TI - Fatal fetal outcome with the combined use of valsartan and atenolol. AB - OBJECTIVE: To report a case of anhydramnios, pulmonary hypoplasia, very small placenta, and fetal death in a pregnancy complicated by chronic hypertension and diabetes mellitus that had been treated through the first 24 weeks of gestation with valsartan and atenolol. CASE SUMMARY: A 40-year-old Hispanic woman with well controlled chronic hypertension and diet-controlled type 2 diabetes mellitus was treated with valsartan and atenolol until pregnancy was diagnosed at 24 weeks' gestation. An ultrasound examination revealed normal fetal growth and anatomy but anhydramnios (amniotic fluid index 0). Valsartan was discontinued, and amniotic fluid volume normalized within two weeks. Intrauterine fetal death was documented at 33 weeks' gestation. Labor was induced, with the delivery of a stillbom female fetus with small, hypoplastic lungs (weight 41% of expected) and an extremely small, 148-g placenta (weight 48% of the 10th percentile for gestational age). DISCUSSION: The use of valsartan, a selective angiotensin II receptor antagonist (ARA), in human pregnancy has not been reported, but this class of agents would be expected to cause fetal toxicity similar to that observed with angiotensin converting enzyme inhibitors. This toxicity includes reduced perfusion of the fetal kidneys, resulting in anuria, oligohydramnios, and subsequent pulmonary hypoplasia. The small hypoplastic lungs and very small placenta were probably a consequence of valsartan and atenolol combination therapy. CONCLUSIONS: Resolution of anhydramnios after discontinuing valsartan is evidence for ARA induced fetal toxicity. The pulmonary hypoplasia observed in the stillbom infant was a direct result of the severe oligohydramnios. The cause of fetal death nine weeks later is uncertain, but because the woman's chronic hypertension and diabetes were well controlled, we believe the primary cause was chronic placental insufficiency resulting from the previous combination of valsartan and atenolol. PMID- 11485134 TI - Sudden cardiac death with clozapine and sertraline combination. AB - OBJECTIVE: To report a case of sudden cardiac death in a patient receiving combination therapy with clozapine and sertraline. CASE SUMMARY: A 26-year-old white man was discovered dead at his residence. His medical history included chronic paranoid schizophrenia, obsessive-compulsive disorder, major depressive disorder, obstructive sleep apnea, and akathisia. He had no prior history of cardiovascular disease. His medication regimen included clozapine 100 mg twice daily (started 4 y prior to his death), risperidone 3 mg twice daily, sertraline 200 mg once daily, atenolol 50 mg twice daily, and lorazepam 0.5 mg four times daily. Autopsy and toxicology studies revealed cardiomegaly suggestive of idiopathic cardiomyopathy, single-vessel coronary artery disease, sertraline and clozapine blood concentrations in the expected range, undetectable lorazepam and risperidone blood concentrations, obesity, and moderate fatty changes to the liver. The most likely cause of death was sudden cardiac death due to acute cardiac arrhythmia. DISCUSSION: Clozapine is structurally similar to the tricyclic antidepressants, which have type 1 A antiarrhythmic properties. Case reports have described electrocardiographic abnomalities, cardiomyopathy, and fatal myocarditis associated with its use. Unexplained death in patients on clozapine therapy has also been reported. Sertraline appears to have less cardiac effect; however, one report has observed clinically significant QT prolongation during sertraline therapy. CONCLUSIONS: Clozapine-induced cardiomyopathy and cardiac arrhythmia from clozapine and/or sertraline use may have contributed to this man's death. PMID- 11485135 TI - Failure of tolterodine to treat clozapine-induced nocturnal enuresis. AB - OBJECTIVE: To report the use and subsequent failure of the bladder-selective agent tolterodine, to treat clozapine-induced nocturnal enuresis in an adolescent patient with psychotic illness. CASE SUMMARY: A 16-year-old Hispanic girl was admitted to the state psychiatric hospital with a diagnosis of bipolar disorder with psychotic features. Clozapine therapy was initiated, and after three months of treatment the patient began experiencing episodes of nocturnal enuresis. The bladder-selective agent tolterodine was tried and subsequently failed to resolve the enuresis episodes. Desmopressin was initiated, which resulted in amelioration of symptoms. DISCUSSION: This is the first published report of using tolterodine to treat clozapine-induced nocturnal enuresis. Several methods to decrease clozapine-induced urinary incontinence have been used and typically include the addition of agents with high anticholinergic properties. Tolterodine is a bladder selective anticholinergic agent indicated for the treatment of urinary urge incontinence and may be employed as a treatment for antipsychotic-induced incontinence. CONCLUSIONS: Nocturnal enuresis is an adverse effect that infrequently occurs with use of clozapine therapy. Although tolterodine was ineffective in our patient to treat clozapine-induced nocturnal enuresis, further trials are required to appropriately evaluate the effectiveness of tolterodine to treat this adverse drug reaction. PMID- 11485136 TI - Serotonin syndrome: early management with cyproheptadine. AB - OBJECTIVE: To report a psychiatric patient who developed serotonin syndrome after a medication overdose and whose marked mydriasis was quickly reversed by administration of cyproheptadine. This phenomenon was confirmed when other cases of serotonin syndrome were studied. METHOD: In the index patient as well as in three subsequent cases of serotonin syndrome, pupil diameter, muscle tone, mental status, and vital signs were monitored before and after a test dose of cyproheptadine as medications were discontinued and antiserotonergic therapy begun. RESULTS: In each patient, cyproheptadine produced rapid reversal of mydriasis within one hour of the initial dose. Other signs of serotonin syndrome remitted more slowly. As the signs and symptoms of serotonin syndrome remitted and pupils returned to normal size and reactiveness, cyproheptadine therapy seemed to produce mydriasis after each dose. Cessation of therapy after this point did not result in recurrence of symptoms. One patient developed serotonin syndrome twice. Two patients developed serotonin syndrome during treatment with medications that are partial serotonin antagonists (mirtazapine and nefazodone). CONCLUSIONS: Rapid reversal of mydriasis in serotonin syndrome by cyproheptadine may serve as a specific suppressive test for the condition, and possibly may add to our understanding of the syndrome. Treatment with cyproheptadine is not thought to abbreviate the illness, but provides symptomatic relief while symptoms persist. PMID- 11485137 TI - Burning mouth syndrome after taking clonazepam. AB - OBJECTIVE: To report the first published case of clonazepam-induced burning mouth syndrome (BMS). CASE SUMMARY: A 52-year-old white woman presented to the clinic with burning mouth symptoms. The patient was previously maintained on alprazolam therapy for anxiety, but was switched to clonazepam because of increased anxiety and panic. Clonazepam significantly relieved her symptoms, but after four weeks of therapy, she reported a constant, mild, oral burning sensation. An oral examination was negative for mucosal abnormalities, and laboratory tests were unremarkable. The clonazepam dose was reduced, and the symptoms decreased, but remained intolerable. Clonazepam was discontinued, and the burning mouth symptoms completely resolved. Since no other medications relieved the anxiety and panic symptoms, the patient requested clonazepam to be reinitiated, but she again developed intolerable burning mouth symptoms. As clonazepam was discontinued, the symptoms resolved. DISCUSSION: The clinical presentation of BMS includes burning and painful sensations of the mouth in the absence of mucosal abnormalities. Candidiasis, anemia, menopause, diabetes mellitus, medications, anxiety, and depression are some causes of this syndrome. Paradoxically, clonazepam has been studied for the treatment of BMS and has demonstrated mild to moderate improvement. In this patient, underlying causes of BMS were eliminated when possible. The association between clonazepam and BMS was highly probable according to the Naranjo probability scale. CONCLUSIONS: This is the first published report describing BMS with a benzodiazepine. Although uncommon, clinicians should be aware of this potential adverse effect due to the widespread use of benzodiazepines. PMID- 11485138 TI - Metformin in an HIV-infected patient with protease inhibitor-induced diabetic ketoacidosis. AB - OBJECTIVE: To describe a case of diabetes mellitus and diabetic ketoacidosis in a patient receiving protease inhibitor therapy and to describe the patient's response to treatment with metformin. CASE SUMMARY: A 49-year-old HIV-positive white man who was receiving indinavir, stavudine, and lamivudine for more than two years presented with shortness of breath and significant weight loss over the previous month. On admission, he had a pH of 7.11 and PaCO2 of 12.9 mm Hg. Laboratory investigations revealed glucose 420 mg/dL, a total carbon dioxide 5 mEq/L, and anion gap of 32. Beta-hydroxybutyrate was 5.9 mmol/L (normal value <0.4 mmol/L). Urine was highly positive for glucose and ketones. The patient was given intravenous fluids and an insulin infusion was started. Five days later, he was discharged on 60 units of insulin per day. Following discharge, efavirenz was substituted for indinavir. Metformin was added and six months following discharge the patient's blood glucose was well controlled with 36 units of insulin per day. DISCUSSION: New-onset diabetes mellitus has been reported in HIV-infected patients receiving protease inhibitors. To date, diabetic ketoacidosis has been an infrequent acute complication. The mechanism by which protease inhibitors cause diabetes is unclear; however, studies have noted insulin resistance and increased proinsulin. Metformin increases the sensitivity of peripheral tissues to insulin and appeared to be useful in this patient. However, further clinical research is needed. CONCLUSIONS: Monitoring glucose concentrations in HIV positive patients receiving protease inhibitors is important to prevent the development of acute complications, including diabetic ketoacidosis. We recommend that these patients have their fasting serum glucose concentration measured at baseline, with follow-up every three months. The role of metformin and the thiazolidinedione antidiabetic agents in the management of protease inhibitor induced diabetes requires further study. PMID- 11485139 TI - Recurrent ingrown toenails secondary to indinavir/ritonavir combination therapy. AB - OBJECTIVE: To report five cases of ingrown toenails (IGTN) associated with indinavir/ritonavir (IDV/RTV) combination therapy. CASE SUMMARY: The median onset of IGTN from initiation of IDV/RTV therapy was 18.4 weeks. Four patients previously received IDV, with one of these experiencing prior IGTN. All patients required surgical management of IGTN. All patients received virologic benefit from ongoing antiretroviral therapy, and the majority of patients elected to maintain IDV/RTV combination therapy. Two patients experienced recurrent IGTN while receiving ongoing IDV/RTV combination therapy. DISCUSSION: IGTN and paronychia have previously been reported with IDV and lamivudine. IGTN in patients with HIV infection is more likely to present acutely, involve more digits, and require surgical management IDV increases retinoic acid signaling and, based on elevated IDV concentrations from concomitant RTV therapy, the risk of IGTN may be increased in patients receiving IDV/RTV combination therapy. CONCLUSIONS: With the increasing popularity of IDV/RTV combination therapy, clinicians should be aware of the potential increase in frequency of dose-related toxicities including IGTN. Evaluation of hands and feet on physical examination should be recommended for all patients being treated with lamivudine and IDV, especially when used in combination with RTV. PMID- 11485140 TI - Use of lepirudin in patients with heparin-induced thrombocytopenia and renal failure requiring hemodialysis. AB - OBJECTIVE: To report two cases of successful lepirudin use in two patients with heparin-induced thrombocytopenia (HIT) and renal failure. CASE SUMMARY: Two patients with renal failure requiring hemodialysis developed HIT syndrome during intravenous heparin therapy. Anticoagulation was necessary to prevent recurrent, acute venous thrombosis in one patient and to prevent arterial thrombosis associated with the use of an intraaortic balloon pump in the second. Intravenous lepirudin was initiated at doses of 0.01 mg/kg/h and 0.005 mg/kg/h, respectively, and titrated based on the activated partial thromboplastin time (aPTT). Steady state doses were 0.015 mg/kg/h to maintain aPTT values of approximately 60 seconds in one patient, and 0.005-0.008 mg/kg/h to achieve an aPTT of approximately 45 seconds in the other patient. DISCUSSION: Lepirudin is one of few anticoagulants that can be safely used in patients with HIT. Because it is eliminated through the kidneys, great care must be taken when administering lepirudin to patients with renal failure; in fact, its use is currently not recommended in patients requiring hemodialysis. Lepirudin effectively prevented acute thrombosis in both of our patients with documented HIT, with no bleeding complications. We describe how we selected the initial doses and report results of aPTT monitoring. CONCLUSIONS: In patients with renal failure who develop HIT, lepirudin is one available alternative to heparin despite its poor renal elimination pattern and subsequently prolonged half-life. PMID- 11485141 TI - Linear immunoglobulin A bullous dermatosis induced by gemcitabine. AB - OBJECTIVE: To report a case of linear immunoglobulin (Ig) A bullous dermatosis (LABD) induced by gemcitabine. CASE SUMMARY: A 59-year-old man was diagnosed with squamous-cell carcinoma of the lung in T4N2M0 stage and treated with cisplatin, vinorelbine, and gemcitabine. Twenty-four hours after the administration of gemcitabine, a symmetric, bullous, herpetiform eruption appeared on his trunk and upper limbs. Histopathologic examination and direct immunofluorescence test were consistent with IgA bullous dermatosis. Cutaneous lesions resolved two weeks after the drug was withdrawn and topical steroid treatment was instituted. DISCUSSION: Drug-induced LABD is a variant of classic or idiopathic LABD. Vancomycin is the most frequently implicated drug, but other agents have been reported to cause LABD. According to the Naranjo probability scale, the relationship of gemcitabine treatment with cutaneous eruption in our patient is possible. CONCLUSIONS: We report the first case of gemcitabine-induced LABD. Clinicians should monitor patients receiving this drug for signs of LABD. PMID- 11485142 TI - Pulmonary fibrosis induced by cyclophosphamide. AB - OBJECTIVE: To report a case of pulmonary fibrosis resulting from use of cyclophosphamide as chemotherapy to treat a patient with breast cancer. CASE SUMMARY: We describe the case of a 52-year-old woman with breast cancer who developed pulmonary fibrosis after four cycles of chemotherapy that included cyclophosphamide. Pulmonary function tests revealed the presence of a severe ventilatory restriction. The open lung biopsy revealed pulmonary fibrosis with vascular sclerosis and signs of pulmonary hypertension. DISCUSSION: Cyclophosphamide is an alkylating agent that has been associated with interstitial pneumonia and pulmonary fibrosis. The frequency of these unwanted effects is <1%. The clinical picture consists of the progressive appearance of dyspnea and a non-productive cough that progresses to severe pulmonary insufficiency. The risk factors described for these complications have been the use of chemotherapy regimens that include other drugs with known pulmonary toxicities, the cumulative total dose, the addition of radiotherapy, and the use of high doses of cyclophosphamide. CONCLUSIONS: Even though the frequency of pulmonary fibrosis in patients treated with cyclophosphamide-based chemotherapy regimens is low, the presence of dyspnea and an interstitial pattern in a patient makes it necessary to consider that possible drug toxicity. The open lung biopsy is the most accurate diagnostic technique for these cases. The discontinuation of cyclophosphamide and treatment with corticosteroids is usually followed by clinical recovery in approximately 50% of patients and, in some cases, reversal of the lung injury. PMID- 11485143 TI - Colesevelam hydrochloride: a novel bile acid-binding resin. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, efficacy, and adverse effects of colesevelam hydrochloride, a bile acid-binding resin. METHODS: MEDLINE searches (1966-June 2000) and manufacturer prescribing literature were employed to find articles on colesevelam. Additional studies and abstracts were identified from the bibliographies of reviewed literature. STUDY SELECTION AND DATA EXTRACTION: All articles identified from data sources were evaluated, and all information deemed relevant was included in this review. Priority was given to randomized, double-blind, placebo-controlled studies. FINDINGS: Colesevelam HCl is a nonabsorbed hydrogel with bile acid sequestrant properties. Monotherapy using colesevelam in once-daily or two divided daily doses of 1.5-4.5 g has produced significant reductions in total cholesterol and low-density lipoprotein (LDL) cholesterol. Mean LDL cholesterol decreases to 20% have been noted when the patient is on 3.75-4.5 g/d. Increases in high-density lipoprotein (HDL) cholesterol have been observed (up to 9%), whereas triglycerides (TG) have increased significantly to 25% in some studies. In unpublished studies, combined use of colesevelam plus hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitor have produced greater reductions in LDL cholesterol than either the statin or colesevelam administered alone. The efficacy of colesevelam monotherapy is slightly less than or similar to cholestyramine or colestipol in decreasing LDL cholesterol, although colesevelam is more potent on a gram-to-gram basis. Adverse effects have been minimal with colesevelam in published studies; this suggests an advantage over cholestyramine or colestipol therapy. Colesevelam appears to be more cost-effective than the packet dosage form of the brand formulation of the older bile acid resins. Care in selection of an appropriate agent should be exercised when considering the issues of adverse effects and palatability. CONCLUSIONS: Colesevelam alone or combined with an HMG-CoA reductase inhibitor is effective in the reduction of total and LDL cholesterol. Since colesevelam is formulated as a tablet, problems with palatability such as with the powder formulation of the bile acid-binding resins are likely to be eliminated. PMID- 11485144 TI - Statin-fibrate combination therapy. AB - BACKGROUND: Precautionary warnings for severe myopathy and rhabdomyolysis from the coadministration of statins and fibrates have been well publicized. However, a recent cerivastatin labeling change made the combined use with fibric acid derivatives a contraindication. Practical recommendations for clinicians who care for patients with refractory mixed hyperlipidemia are needed. OBJECTIVE: To provide recommendations for clinicians in the treatment of refractory mixed hyperlipidemia. DATA SOURCES: A comprehensive MEDLINE (1966-July 2000) and bibliographic search was performed. DATA SYNTHESIS: Thirty-six published clinical trials and 29 case reports involving combination therapy with hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors and fibric acid derivatives regarding the occurrence of rhabdomyolysis or myopathy were reviewed. The literature review demonstrated that combination therapy with a statin and fibrate increases the risk of muscle damage, with an incidence of 0.12%. Risk factors that predispose patients to myopathy caused by combination statin-fibrate therapy include increased age, female gender, renal or liver disease, diabetes, hypothyroidism, debilitated status, surgery, trauma, excessive alcohol intake, and heavy exercise. CONCLUSIONS: Combination therapy with a statin and fibrate offers significant therapeutic advantage for the treatment of severe or refractory mixed hyperlipidemia. Although such a combination does increase the risk of myopathy, with an incidence of approximately 0.12%, this small risk of myopathy rarely outweighs the established morbidity and mortality benefits of achieving lipid goals. Nevertheless, a higher incidence of myopathy has been reported with statin monotherapy. When monotherapy with a statin fails to control mixed hyperlipidemia, combination therapy may be considered. Niacin may be added before a fibrate is considered, as it appears to have less risk of myopathy. Statin-fibrate combination therapy must be undertaken cautiously and only after careful risk-benefit analysis. Patient counseling on the risks and warning signs of myopathy is extremely important. PMID- 11485145 TI - Effect of common over-the-counter medications on blood alcohol levels. AB - OBJECTIVE: To assess the clinical and legal significance of the potential pharmacokinetic interaction between common over-the-counter (OTC) medications and alcohol that may result in increased blood alcohol levels (BALs). DATA SOURCES: A MEDLINE search (1966-February 2000) of English-language articles was performed using the terms aspirin, acetaminophen, histamine (H2)-receptor antagonist, ethanol, and blood alcohol level and then supplemented by a bibliographic review of relevant articles. STUDY SELECTION AND DATA EXTRACTION: Two H2-receptor antagonist studies using methodologies representative of other published trials and a meta-analysis of 24 H2-receptor antagonist trials were chosen for detailed review. All identified studies examining aspirin and acetaminophen were addressed. DATA SYNTHESIS: More than 30 studies have examined the potential interaction between OTC drugs and blood alcohol. Because this issue has important medical and legal implications for patients, prescribing physicians, and pharmaceutical manufacturers, a critical analysis of the literature addressing this potential interaction is presented. CONCLUSIONS: Numerous factors arguing against a clinically significant interaction were identified. First, data from the relevant studies cannot be extrapolated to the general population because of the multitude of variables that determine an individual's BAL. Also, a publication bias for small studies (< or = 10 subjects) finding a statistically significant increase in peak BAL was observed. In addition, study results supporting an increase in BAL were often irreproducible when these trials were repeated under similar conditions. Finally, although some studies detected statistically significant increases in peak BAL, these changes were often clinically irrelevant. PMID- 11485146 TI - Systemic hemostatic medications for reducing surgical blood loss. AB - OBJECTIVE: To review randomized trials involving the use of systemic hemostatic medications for reducing surgical blood loss. DATA SOURCES: Articles were obtained through searches of MEDLINE (1966-September 2000). The bibliographies of retrieved publications were reviewed for additional references. STUDY SELECTION: All randomized studies and pharmacoeconomic evaluations that involved medications used for systemic hemostasis in the perioperative period were included. DATA EXTRACTION: Randomized studies involving conjugated estrogens, aminocaproic acid, tranexamic acid, desmopressin, and aprotinin for systemic hemostasis were extracted. Studies of proton-pump inhibitors for upper gastrointestinal bleeding and octreotide for variceal bleeding were excluded, as were trials involving the use of any hemostatic agent for cardiovascular surgery. The primary outcome under review was a reduction in bleeding as defined by reduced transfusion requirements. DATA SYNTHESIS: There is limited efficacy and toxicity information concerning the use of conjugated estrogens for reducing surgery-related bleeding. Similarly, there are a limited number of randomized studies involving aminocaproic acid and tranexamic acid, and with the exception of tranexamic acid for reducing transfusion requirements with knee surgery, the study results are either conflicting or negative. For desmopressin, evidence from a substantial number of randomized trials documents its lack of efficacy. Aprotinin has reduced bleeding and transfusion requirements in a number of randomized studies involving patients undergoing orthopedic surgery, but cost-effectiveness studies are needed to better define its therapeutic role. Trials of aprotinin during hepatic surgery have yielded conflicting results. CONCLUSIONS: Most hemostatic medications used for reducing surgery-related bleeding have limited or contradictory evidence of efficacy. PMID- 11485147 TI - Prevention and treatment of veno-occlusive disease. AB - OBJECTIVE: To review the current options for prevention and treatment of veno occlusive disease in bone marrow transplant patients. DATA SOURCES: Articles were selected from a MEDLINE search (1966-October 1999) using the key terms veno occlusive disease and bone marrow transplantation. In addition, references of all articles were examined for articles not found in the computer-based search. DATA EXTRACTION: All clinical trials, case-control studies, and case reports were evaluated. RESULTS: Heparin, low-molecular-weight heparin, prostaglandin E1, ursodiol, and glutamine have been studied for prevention of veno-occlusive disease. Heparin has been studied most extensively; however, no preventive regimen has a defined role in therapy. For treatment, tissue plasminogen activator has been evaluated most thoroughly, yet its safety and efficacy have not been clearly established in patients with veno-occlusive disease. Other possible treatment options include antithrombin-III, defibrotide, glutamine plus vitamin E, and surgery. CONCLUSIONS: Based on the available data, the most promising agents are ursodiol for prevention and defibrotide or glutamineplus vitamin E for treatment of veno-occlusive disease. Further clinical trials are needed to establish the appropriate preventive and treatment options available for bone marrow transplant patients suffering from veno-occlusive disease. To date, such decisions depend largely on poorly designed trials, case reports, and clinical experience. PMID- 11485148 TI - Topical tacrolimus in the treatment of atopic dermatitis. AB - OBJECTIVE: To review the efficacy of topical tacrolimus in the treatment of atopic dermatitis (AD). DATA SOURCES: Searches of MEDLINE (1966-October 2000), International Pharmaceutical Abstracts (1970-October 2000), and ScienceDirect (1994-October 2000) were performed using the key search terms tacrolimus, FK506, and atopic dermatitis. DATA SYNTHESIS: Since patients with AD have defects in cell-mediated immunity, the immunosuppressant properties of the macrolides (cyclosporine and tacrolimus) may prove to be beneficial in the treatment of AD. Topical tacrolimus has been frequently studied in the treatment of AD because it was found to have better skin penetration and higher potentency than topically applied cyclosporine. Studies evaluating the use of topical tacrolimus are presented and provide evidence that topical tacrolimus is effective in the treatment of AD with no evidence thus far of systemic adverse effects. CONCLUSIONS: There is a fair amount of documentation of the efficacy and safety of topical tacrolimus. Further trials are needed to determine the optimal duration of therapy and its efficacy and safety in children less than seven years of age. PMID- 11485149 TI - Sucrose analgesia for minor procedures in newborn infants. AB - OBJECTIVE: To evaluate the role of intraoral sucrose and other sweet-tasting solutions for the management of pain associated with minor procedures in newborns. DATA SOURCES: A search of MEDLINE articles from 1966 to August 1999 and an extensive review of journals was conducted. MeSH headings included analgesia, sucrose, and neonate. DATA SYNTHESIS: Newborn infants regularly undergo minor invasive procedures for which analgesics are not routinely used. Intraoral sucrose and other sweet-tasting solutions appear to diminish surrogate biomarkers of pain response as evidenced by a reduction in crying time, smaller increases in heart rate, and lower pain scale ratings. CONCLUSIONS: These studies appear to suggest that sucrose solution 0.5 g provides effective analgesia with no reported adverse effects. PMID- 11485151 TI - Alternative, complementary, or integrated pharmacotherapy. PMID- 11485150 TI - Evaluation of a new integrated discharge prescription form. AB - OBJECTIVE: To determine whether a new discharge prescription form which integrates admission medications, in-hospital changes, and discharge medications could enhance the accuracy of information in patient profiles in community pharmacies after hospital discharge. DESIGN: Nonrandomized, prospective, multi site study. SETTINGS: Internal medicine wards of the three teaching hospitals (1200 beds) of the Centre Hospitalier de l'Universite de Montreal. SUBJECTS: Patients admitted to the internal medicine wards between January 4 and 31, 1999, at St.-Luc and Notre-Dame Hospitals formed the control group and received a usual discharge form (UD). Those admitted between February 1 and 28,1999, received the new discharge prescription form (DPF) capturing the list of admission medications and revisions during hospitalization; they served as the experimental group. METHODS: Patient profiles were reviewed to calculate conformity rates of community pharmacy patient profiles after discharge and the rate of overall conformity for each group in the study. Each drug in the patient profile was assessed according to six criteria. Healthcare providers' satisfaction with the DPF was assessed via a written questionnaire. RESULTS: Eighty-nine patients and 669 discharge medications were studied. The patient profiles had a higher conformity rate in the DPF group than in the UD group (82% vs. 40%; p < 0.001); improvement could be attributed to higher conformity rates, particularly for two criteria (medications stopped in hospital and dose changes in hospital). CONCLUSIONS: Integration of admission medications, in-hospital changes, and discharge medications on a single form increases the conformity rates of community pharmacy patient profiles after hospitalization. This tool is well accepted by both pharmacists and physicians and may lead to a major decrease in drug-related problems. PMID- 11485152 TI - Lithium-induced exacerbation of stutter. PMID- 11485153 TI - Severe acne with lithium. PMID- 11485154 TI - Homicidal ideation secondary to interferon. PMID- 11485155 TI - BCG therapy-related death and previous pelvic radiation. PMID- 11485156 TI - Polycythemia in a patient treated with isotretinoin. PMID- 11485157 TI - Plasma cholesteryl ester transfer protein and lipoprotein levels during treatment of growth hormone-deficient adult humans. AB - The incidence of atherosclerosis is increased in growth hormone (GH) deficient individuals. Nonetheless, the antiatherogenic benefits of GH replacement therapy remain uncertain. In this study the effect of human recombinant growth hormone (hrGH) replacement therapy administered to GH-deficient adults on the plasma cholesteryl ester transfer protein (CETP) concentration and activity was analyzed. These findings were related to changes in the concentrations of the plasma lipoproteins. The hrGH was administered for 12 mon to human GH-deficient patients (n = 13; 8 men, 5 women). During the study plasma lipoproteins were separated by ultracentrifugation, and plasma cholesterol esterification rate (CER), endogenous CETP activity, and CETP concentration were measured. GH replacement therapy transiently (at 3 mon) lowered plasma concentration of CETP and low density lipoprotein-cholesterol (LDL-C) and raised total triglycerides. Furthermore, hrGH permanently increased both the plasma lipoprotein(a) [Lp(a)] concentration, which is known as atherogenic, and the proportion of cholesteryl ester in the high density lipoprotein2 (HDL2) particles, which is potentially atheroprotective. The simultaneous decrease of the plasma CETP and LDL-C concentrations elicited by hrGH indicated a close relationship between LDL metabolism and the regulation of the CETP gene expression. Endogenous CETP activity and the CER were not modified because these parameters are regulated in opposite ways by plasma levels of triglycerides; that is, CER increased and CETP decreased. PMID- 11485158 TI - Influence of formulas with borage oil or borage oil plus fish oil on the arachidonic acid status in premature infants. AB - Several studies have reported that feeding gamma-linolenic acid (GLA) has resulted in no increase in arachidonic acid (AA) in newborns. This result was ascribed to the eicosapentaenoic acid (EPA)-rich fish oil used in these formulas. Docosahexaenoic acid (DHA) sources with only minor amounts of EPA are now available, thus the addition of GLA to infant formulas might be considered an alternative to AA supplementation. Sixty-six premature infants were randomized to feeding one of four formulas [ST: no GLA, no long-chain polyunsaturated fatty acids; BO: 0.6% GLA (borage oil); BO + FOLOW: 0.6% GLA, 0.3% DHA, 0.06% EPA; BO + FOHIGH: 0.6% GLA, 0.3% DHA, 0.2% EPA] or human milk (HM, nonrandomized) for 4 wk. Anthropometric measures and blood samples were obtained at study entry and after 14 and 28 d. There were no significant differences between groups in anthropometric measures, tocopherol, and retinol status at any of the studied time points. The AA content of plasma phospholipids was similar between groups at study start and decreased significantly until day 28 in all formulafed groups, but not in the breast-fed infants [ST: 6.6 +/- 0.2%, BO: 6.9 +/- 0.3%, BO + FOLOW: 6.9 +/- 0.4%, BO + FOHIGH: 6.7 +/- 0.2%, HM: 8.6 +/- 0.5%, where values are reported as mean +/- standard error; all formulas significantly different (P< 0.05) from HM]. There was no significant influence of GLA or fish oil addition to the diet. GLA had only a very limited effect on AA status which was too small to obtain satisfactory concentrations (concentrations similar to breast-fed babies) under the circumstances tested. The effect of GLA on AA is independent of the EPA and DHA content in the diet within the dose ranges studied. PMID- 11485159 TI - Effects of delta5 polyunsaturated fatty acids of maritime pine (Pinus pinaster) seed oil on the fatty acid profile of the developing brain of rats. AB - Conifer (pine) seeds are a potential source of dietary oils, but their safety and nutritional properties are not well established. Conifer seed oils differ from common edible vegetable oils in having a series of unusual polyunsaturated fatty acids (PUFA) with a polymethylene-interrupted (PMI) double bond system and a double bond at the delta5 position. A rat study was conducted to assess whether delta5 PMI-PUFA of conifer seeds could alter the levels of n-6 and n-3 long-chain polyunsaturated fatty acids (LC-PUFA) in mothers' milk and the developing brain of fetuses and pups. Feeding maritime pine (Pinus pinaster) seed oil (MPO) diet with a delta5 PMI-PUFA content of 1.4 g/100 g throughout pregnancy and lactation resulted in a large incorporation of delta5 PMI-PUFA in mothers' milk (5.1 +/- 0.5% of total fatty acids). The fetus (17 d old) and pup (22 d) brains, however, accumulated very little (0.6 and 0.4% of total fatty acids, respectively) delta5 PMI-PUFA. Mother's milk and pup's brain of the MPO group contained normal levels of 20:4n-6, 22:4n-6, and 20:5n-3 compared to a reference group of rats fed a fat blend of sunflower, high-oleic sunflower, and canola oils. The level of 22:6n-3, however, was slightly but significantly (P < 0.05) higher in milk and pup brain of the MPO group. These results show that delta5 PMI-PUFA of MPO exert no negative effect on the levels of n-6 and n-3 LC-PUFA in rat brain during its early development. PMID- 11485160 TI - The effect of conjugated linoleic acid isomers on fatty acid profiles of liver and adipose tissues and their conversion to isomers of 16:2 and 18:3 conjugated fatty acids in rats. AB - Conjugated linoleic acid (CLA) is a collective term that describes different isomers of linoleic acid with conjugated double bonds. Although the main dietary isomer is 9cis,11trans-18:2, which is present in dairy products and ruminant fat, the biological effects of CLA generally have been studied using mixtures in which the 9cis,11trans- and the 10trans,12cis-18:2 were present at similar levels. In the present work, we have studied the impact of each isomer (9cis,11 trans- and 10trans,12cis-18:2) given separately in the diet of rats for 6 wk. The 10trans,12cis-18:2 decreased the triacylglycerol content of the liver (-32%) and increased the 18:0 content at the expense of 18:1 n-9, suggesting an alteration of the delta9 desaturase activity, as was already demonstrated in vitro. This was not observed when the 9cis,11trans-18:2 was given in the diet. Moreover, the 10trans,12cis-18:2 induced an increase in the C22 polyunsaturated fatty acids in the liver lipids. The 10trans,12cis-18:2 was mainly metabolized into conjugated 16:2 and 18:3, which have been identified. The 9cis,11trans isomer was preferentially metabolized into a conjugated 20:3 isomer. Thus, the 9cis,11trans- and the 10trans,12cis-CLA isomers are metabolized differently and have distinct effects on the metabolism of polyunsaturated fatty acids in rat liver while altering liver triglyceride levels differentially. PMID- 11485161 TI - A single oral administration of conjugated linoleic acid enhanced energy metabolism in mice. AB - We investigated the effect of a single oral administration of conjugated linoleic acid (CLA) on energy metabolism in mice. Male Std ddY mice were orally administered CLA (5 mL/kg weight) or linoleic acid (5 mL/kg weight) (both solutions at concentrations of 73.5%) as a control. Oxygen consumption was significantly greater in the CLA-administered mice than in the control mice. Respiratory quotient was slightly lower in the CLA-adminis-tered mice than in the control mice. We calculated fat and carbohydrate oxidation from oxygen consumption and respiratory quotient. Fat oxidation in the CLA-administered mice was significantly higher than in the control mice, and there was no difference in carbohydrate oxidation. Serum concentrations of noradrenalin and adrenalin in the CLA administered mice were significantly higher than in the control mice. These results suggested that CLA enhanced sympathetic nervous activity and energy metabolism. PMID- 11485162 TI - Effect of dietary restriction on age-related increase of liver susceptibility to peroxidation in rats. AB - Dietary restriction (DR) increases life span and decreases age-related diseases in experimental animals. It has received a great deal of attention in connection with the relationship between aging, nutrition, and oxidative stress because oxidative injury in several organ systems is a prominent feature in aging. We investigated the possibility that DR can protect vulnerable liver lipids against age-related increases of peroxidation. Male Fischer 344 rats fed ad libitum (AL) or dietarily restricted (maintained on 60% of AL food intake) were killed by decapitation at 4 (young) or 12 mon (adult) of age. Phosphatidylcholine hydroperoxide (PCOOH) concentration of liver was determined using a chemiluminescent high-performance liquid chromatographic method. Liver PCOOH increased with age in adult rats, but less of an increase of PCOOH was seen in DR rats, which is consistent with results on production of thiobarbituric acid reactive substances and oxygen-derived free radicals. No significant differences were found in liver superoxide dismutase and catalase activity between AL and DR groups of young and adult rats. Liver triglyceride and cholesterol contents were lower in DR than AL rats at 12 mon. Fatty acid compositions of phosphatidylcholine and phosphatidylethanolamine indicated that the ratio of (20:3n-6 + 20:4n-6)/18:2n-6, an index of linoleic acid (18:2n-6) desaturation, was lower in DR than in AL rats. We concluded that DR suppresses age-related oxidative damage in liver by modulating the amount of lipid as well as fatty acid composition. PMID- 11485163 TI - Overexpression of acyl-coA binding protein and its effects on the flux of free fatty acids in McA-RH 7777 cells. AB - Overexpression of acyl-CoA binding protein (ACBP) was induced in a rat hepatoma cell line (McA-RH 7777) by stable integration of rat ACBP cDNA. The transfected cells (ACBP-27) had 3.5-fold higher concentrations of ACBP than control cells (14 vs. 4 ng/microg DNA). Both ACBP-27 and control cells were cultured in the presence of various concentrations of radiolabeled palmitic acid; and the effects of ACBP on lipogenesis and beta-oxidation were studied. Incubation of the cells with 100 microM palmitic acid resulted in 42% greater incorporation of the fatty acid in ACBP-27 cells as compared to that in the control cells. This increased incorporation of the fatty acid was observed predominantly in the triglyceride fraction. Higher concentrations of palmitic acid (200 to 400 microM) were associated with a significant decrease in the production of 14CO2 in the ACBP-27 cell line than in the control cells, while lower concentrations had no effect. Our data suggest a role for ACBP in the partitioning of fatty acids between esterification reactions leading to the formation of neutral lipids and beta oxidation. ACBP may play a regulatory role by influencing this important branch point in intermediary lipid metabolism. PMID- 11485164 TI - Metabolism of very long chain polyunsaturated fatty acids in isolated rat germ cells. AB - Which cell type is responsible for the high levels of very long chain polyunsaturated fatty acids in testis and whether this fatty acid pattern is a result of a local synthesis are not presently known. In this study, fatty acid conversion from 20:4n-6 to 22:5n-6 and from 20:5n-3 to 22:6n-3 was investigated in isolated rat germ cells incubated with [1-14C]-labeled fatty acids. The germ cells elongated the fatty acids from 20- to 22-carbon atoms and from 22- to 24 carbon atoms but had a low delta6 desaturation activity. Thus, little [14C]22:5n 6 and [14C]22:6n-3 were synthesized. When Sertoli cells were incubated with [1 14C]20:5n-3 for 24 h, an active fatty acid elongation and desaturation were observed. In vivo germ cells normally have a higher content of 22:5n-6 or 22:6n-3 than Sertoli cells. An eventual transport of essential fatty acids from Sertoli cells to germ cells was thus studied. Different co-culture systems were used in which germ cells were on one side of a filter and Sertoli cells on the opposite side. When isolated pachytene spermatocytes or round spermatids were added to the opposite side of a semipermeable filter, approximately 1 nmol [14C]22:6n-3 crossed the filter. Little of this was esterified in the germ cells. Similarly, in using [1-14C]20:4n-6 in identical experiments, very little [14C]22:5n-6 was esterified in germ cells on the opposite side of the filter. Although the very active synthesis of 22:5n-6 and 22:6n-3 observed in Sertoli cells suggests a transport of these compounds to germ cells, this was not experimentally determined. PMID- 11485165 TI - Oleate acutely stimulates the secretion of triacylglycerol by cultured rat hepatocytes by accelerating the emptying of the secretory compartment. AB - The acute effects of addition of oleate on the rate of triacylglycerol (TAG) secretion by cultured rat hepatocytes were studied by monitoring the use of endogenous (14C-prelabeled) acyl moieties and exogenous (3H-labeled) oleate for the synthesis of secreted TAG simultaneously. Inclusion of exogenous oleate in the medium stimulated the secretion of the endogenous 14C-labeled acyl moieties by 55-100%. To find out whether the stimulation was due to increased endogenous TAG mobilization or an increased rate of processing of TAG within the endoplasmic reticulum (ER) secretory machinery, use was made of the inhibition of apolipoprotein B (apoB) synthesis (but not degradation) by Ca2+ mobilization from the ER. Inhibition of apoB synthesis stopped entry of acyl moieties (from endogenous and exogenous sources) into the secretory pathway. However, even when entry of acyl moieties into the secretory pathway was totally inhibited, exogenous oleate was still able to stimulate (twofold) the secretion [14C]TAG, indicating that oleate stimulates the emptying of prelabeled TAG from the secretory compartment at a point distal to apoB synthesis and nascent particle formation. These data indicate that exogenous oleate, besides providing additional acyl moieties for incorporation into secreted TAG, stimulates the secretion of endogenous TAG in a manner (i) that is independent of effects on apoB synthesis and/or degradation and (ii) that involves the enhanced processing of TAG resident within the ER secretory pathway. PMID- 11485166 TI - Necrosis and apoptosis in lymphoma cell lines exposed to eicosapentaenoic acid and antioxidants. AB - The present study is focused on the role of oxidative stress in the induction of either necrosis or apoptosis by eicosapentaenoic acid (EPA) in the lymphoma cell lines Raji and Ramos, respectively. To investigate the different death modes induced by EPA, we assessed the importance of some antioxidants and reactive oxygen species in the two cell lines. We observed that different antioxidants counteracted the necrotic effect of EPA on Raji cells to a different extent, and that vitamin E counteracted EPA-induced accumulation of superoxide anion in this cell line. On the contrary, no effects of antioxidants were observed on development of apoptosis induced by EPA in Ramos cells, and vitamin E did not counteract EPA-induced accumulation of superoxide anions in Ramos cells. Moreover, apoptosis was partly inhibited by transcription inhibitors (actinomycin D) and protein synthesis inhibitors (cycloheximide), suggesting dependency upon new protein synthesis prior to apoptosis. Kinase inhibitors (staurosporin and calphostin C) did not alter the EPA-induced apoptosis. The observed cellular accumulation of superoxide anion following EPA incubation may be important for induction of necrosis in Raji cells. In contrast, none of the other investigated parameters indicated a role of oxidative stress promoted by EPA in the induction of apoptosis in Ramos cells. PMID- 11485167 TI - Effect of 4-hydroxy-2(E)-nonenal on soybean lipoxygenase-1. AB - The oxidation of linoleic acid by soybean lipoxygenase-1 (LOX-1) was inhibited in a time-dependent manner by 4-hydroxy-2(E)-nonenal (HNE). Kinetic analysis indicated the effect was due to slow-binding inhibition conforming to an affinity labeling mechanism-based inhibition. After 25 min of preincubation of LOX-1 with and without HNE, Lineweaver-Burk reciprocal plots indicated mixed noncompetitive/competitive inhibition. Low concentrations of HNE influenced the electron paramagnetic resonance (EPR) signal of 13(S)-hydroperoxy-9(Z), 11 (E) octadecadienoic acid (13-HPODE)-generated Fe3+-LOX-1 slightly, but higher concentrations completely eliminated the EPR signal indicating an active site hindered from access by 13-HPODE. HNE may compete for the active site of LOX-1 because its precursor, 4-hydroperoxy-(2E)-nonenal, is a product of LOX-1 oxidation of (3Z)-nonenal. Also, it was an attractive hypothesis to suggest that HNE may disrupt the active site by forming a Michael adduct with one or more of the three histidines that ligate the iron active site of LOX-1. PMID- 11485168 TI - Vinyl sulfide derivatives of truncated oxidosqualene as selective inhibitors of oxidosqualene and squalene-hopene cyclases. AB - Various vinyl sulfide and ketene dithioacetal derivatives of truncated 2,3 oxidosqualene were developed. These compounds, having the reactive functions at positions C-2, C-15 and C-19 of the squalene skeleton, were studied as inhibitors of pig liver and Saccharomyces cerevisiae oxidosqualene cyclases (OSC) (EC 5.4.99.7) and of Alicyclobacillus acidocaldarius squalene hopene cyclase (SHC) (EC 5.4.99.-). They contain one or two sulfur atoms in alpha-skeletal position to carbons considered to be cationic during enzymatic cyclization of the substrate and should strongly interact with enzyme nucleophiles of the active site. Most of the new compounds are inhibitors of the OSC and of SHC, with various degrees of selectivity. The methylthiovinyl derivative, having the reactive group at position 19, was the most potent and selective inhibitor of the series toward S. cerevisiae OSC, with a concentration inhibiting 500% of the activity of 50 nM, while toward the animal enzyme it was 20 times less potent. These results could offer new insight for the design of antifungal drugs. PMID- 11485169 TI - Composition of the silk lipids of the spider Nephila clavipes. AB - A detailed analysis of the lipids of spider silk is given for the first time. Extracts of the silk from the golden orb weaver, Nephila clavipes, were studied by gas chromatography, mass spectrometry, and chemical derivatizations. The major group of the lipids consisted of methyl-branched 1-methoxyalkanes (methyl ethers) with up to four methyl groups in the chain (chain length between C28 and C34), which are unique to spiders. The position of the methyl branches was determined by conversion into cyanides, which allowed easy location of methyl branches. The second-largest group included alkanes with a wide structural variety; 2-methyl branched, even-numbered hydrocarbons predominated. A general numerical method for the estimation of retention indices of alkanes and their derivatives is presented. Further components of the web included alkanols and alkanediols, fatty acids, and glyceryl ethers. Some comments on the biosynthesis of these compounds are also given. PMID- 11485170 TI - Fullerenoid lipids: first synthesis of structured triacylglycerols containing an Aza-[60]fullerene unit. AB - Some 1,2- and 1,3-diacyl glycerols (with acyl groups as stearyl, oleyl, linoleyl, or stearolyl) were synthesized by conventional methods. The diacyl glycerols were esterified with 6-bromo-hexanoic acid to give the corresponding bromo triacylglycerols (of the type AAB and ABA containing a bromo group at the distal part of the hexanoate chain). The bromo function was transformed to an azide group by reaction of the bromo-triacylglycerols with sodium azide. The resulting azido-triacylglycerols were then reacted with [60]fullerene to give the requisite aza-fullerenoid triacylglycerol of the type ABA or AAB (45-62% yield based on the amount of [60]fullerene reacted). The nitrogen atom attached to the carbon cage formed a "[5,6]open" type aza substructure, which was confirmed by the appearance of 31-32 signals in the region of deltaC 133-148 (carbon shifts of Sp2 carbons of the cage) in the 13C nuclear magnetic resonance spectra. The spectroscopic and mass spectrometric properties of these novel fullerenoid triacylglycerols are reported. PMID- 11485171 TI - Influence of chlorpheniramine maleate on topical hydroxypropylcellulose films produced by hot-melt extrusion. AB - The objective of this investigation is to study the influence of chlorpheniramine maleate (CPM) on the chemical and physical-mechanical properties of hydroxypropylcellulose (HPC) hot-melt extruded films without the use of a traditional plasticizer HPC films containing CPM in concentrations of 1, 5, and 10 wt% were prepared by hot-melt extrusion utilizing a Randcastle Microtruder (Model #RCP-0750) with a 6-in. flex-film die. The physical-mechanical properties including tensile strength and percent elongation were determined on an Instron according to the ASTM standards. Glass transition temperatures and thermal analysis of the extruded films were determined utilizing a DSC 2920 Modulated DSC and Thermal Analyst 2000 software. The crystalline properties of the drug, polymer, and extruded films were studied via wide angle X-ray diffraction (XRD) using a Philips Vertical Scanning Diffractometer (Type 42273, Philips Electronic Instrument, Mount Vernon, NY). Gel permeation chromatography was used to study the stability of the polymer matrix as a function of different concentrations of CPM and processing conditions. CPM functioned as an effective plasticizer, increasing percent elongation and decreasing tensile strength in a concentration dependent manner All three concentrations of extruded films exhibited a 10- to 12 fold decrease in tensile strength in contrast to a fourfold increase in percent elongation when testing was performed perpendicular to flow vs. in the direction of flow. The drug was also shown by XRD and DSC data to be in solution in the HPC matrix within the films up to the 10% level. In addition, CPM functioned as a processing aid in the extrusion of hot-melt films, stabilizing the weight-average molecular weight of HPC and allowing for film processing at lower temperatures. CPM could potentially be a candidate antihistamine for transdermal or transmucosal applications in film devices prepared by hot-melt extrusion technology. PMID- 11485172 TI - Enhancement of ibuprofen dissolution via wet granulation with beta-cyclodextrin. AB - The purpose was to investigate the effect of wet granulation with beta cyclodextrin (betaCD) on the enhancement of ibuprofen (IBU) dissolution. The effect of the granulation variables on the physical properties as well as the dissolution of tablets prepared from these granules was also examined. Granulation was performed using three granulating solvents: water, ethanol (95 vol%), and isopropanol. Granules were either oven-dried for 2 h or air-dried for 3 days. The granules or respective physical mixtures were compressed into tablets. Powder X-ray diffraction showed that oven-dried granulation resulted in less amorphous entities thatfacilitated IBU-betaCD complexation in solution and enhanced the dissolution of the corresponding tablets compared to the physical mixture with or without oven drying. In contrast, air-dried granulation did not cause any differences in the X-ray diffraction pattern (crystallinity) or the dissolution compared to the physical mixture without drying. Isopropanol and water, as granulating solvents, enhanced the dissolution of the oven-dried batches more than ethanol. The Differential scanning calorimetry (DSC) and Thermogravimetric analysis (TGA) data showed that tablets prepared from oven dried granules, but not air-dried granules, had lower AH values and percent loss in weight, respectively, than those prepared from the physical mixture as a result of the expulsion of the water molecules from the betaCD cavity and enhancement of the complexation in solution. These results showed that oven-dried granulation of IBU and betaCD provided faster IBU dissolution than the physical mixture; air-dried granulation did not substantially affect the dissolution of IBU. PMID- 11485173 TI - Elucidation of solution state complexation in wet-granulated oven-dried ibuprofen and beta-cyclodextrin: FT-IR and 1H-NMR studies. AB - The effect of oven-dried wet granulation on the complexation of beta-cyclodextrin with ibuprofen (IBU) in solution was investigated using Fourier transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance (1H NMR), and molecular modeling. Granulation was carried out using 5 mL of three different granulating solvents; water, ethanol (95% v/v), and isopropanol and the granules were oven-dried at 60 degrees C for 2 h. The granules were compared to oven-dried physical mixture and conventionally prepared complex. Phase solubility study was performed to investigate the stability of the granulation-formed complexes in solution. FT-IR was used to examine the complexation in the granules while 1H NMR, and molecular modeling studies were carried out to determine the mechanism of complexation in the water-prepared granules. The solubility studies suggested a 1:1 complex between IBU and betaCD. It also showed that the stability of the complex in solution was in the following order with respect to the granulating solvents: ethanol > water > isopropanol. The FT-IR study revealed a shift in the carboxylic acid stretching band and decrease in the intensities of the C-H bending bands of the isopropyl group and the out-of-plane aromatic ring, of IBU, in granules compared to the oven-dried physical mixture. This indicated that granules might have some extent of solid state complexation that could further enhance dissolution and the IBU-betaCD solution state complexation. 1H NMR showed that water prepared oven-dried granules had a different 1H NMR spectrum compared to similarly made oven-dried physical mixture, indicative of complexation in the former. The 1H NMR and the molecular modeling studies together revealed that solution state complexation from the granules occurred by inclusion of the isopropyl group together with part of the aromatic ring of IBU into the betaCD cavity probably through its wider side. These results indicate that granulation process induced faster complexation in solution which enhances the solubility and the dissolution rate of poorly soluble drugs. The extent of complexation in the granules was dependent on the type of solvent used. PMID- 11485174 TI - Influence of plasticization time, curing conditions, storage time, and core properties on the drug release from Aquacoat-coated pellets. AB - Theophylline or chlorpheniramine maleate pellets were coated with an aqueous ethylcellulose dispersion, Aquacoat. The influence of the plasticization time, curing conditions, storage time, and core properties on the drug release were investigated. The plasticization time (time between plasticizer addition to the polymer dispersion and the spraying process) did not affect the drug release, when the water-soluble plasticizer triethyl citrate, was used because of its rapid uptake by the colloidal polymer particles. In contrast, with the water insoluble plasticizer acetyltributyl citrate (ATBC), plasticization time (1/2 h vs 24 h) influenced the drug release, the longer plasticization time resulted in a slower drug release because of a more complete plasticizer uptake prior to the coating step. However a thermal aftertreatment of the coated pellets at eleylated temperatures (curing step) reduced/eliminated the effect of the plasticization time with ATBC. In general, curing reduced the drug release and resulted in stable drug release profiles. The time period between the coating and the curing step was not critical when the pellets were cured for a longer time. The structure of the pellet core (high dose matrix vs low dose layered pellet) strongly affected the drug release. A slow, zero-order drug release was obtained with high dose theophylline pellets, while a more rapid, first-order release pattern was obtained with low dose theophylline-layered nonpareil pellets. PMID- 11485175 TI - An evaluation of three-dimensional modeling of compaction cycles by analyzing the densification behavior of binary and ternary mixtures. AB - The aim of the study is to use the 3D modeling technique of compaction cycles for analysis of binary and ternary mixtures. Three materials with very different deformation and densification characteristics [cellulose acetate (CAC), dicalcium phosphate dihydrate (EM) and theophylline monohydrate (TM)] have been tableted at graded maximum relative densities (rhorel, max) on an eccentric tableting machine. Following that, graded binary mixtures from CAC and EM have been compacted. Finally, the same ratios of CAC and EM have been tableted in a ternary mixture with 20 vol% TM. All compaction cycles have been analyzed by using different data analysis methods. Three-dimensional modeling, conventional determination of the slope of the Heckel function, determination of the elastic recovery during decompression, and calculations according to the pressure-time function were the methods of choice. The results show that the 3D model technique is able to gain the information in one step instead of three different approaches, which is an advantage for formulation development. The results show that this model enables one to better distinguish the compaction properties of mixtures and the interaction of the components in the tablet than 2D models. Furthermore, the information by 3D modeling is more precise since in the slope K of the Heckel-plot (in die) elasticity is included, and in the parameters of the pressure-time function beta and gamma plastic deformation due to pressure is included. The influence of time and pressure on the displacement can now be differentiated. PMID- 11485176 TI - Divisability of diltiazem matrix sustained-release tablets. AB - The objective of this work was to study the possibility of a solid sustained release dosage form, like a tablet, be divided without changing its release characteristics. Diltiazem hydrochloride Sustained-Release (SR) tablets with a standard groove on one face, were tested and the following dissolution parameters were evaluated: t10%, t25%, and t50% dissolution time, and dissolution efficiency at t120, and at t360. To analyze the release mechanism, several release models were tested such as Higuchi, zero order, first order, Baker-Lonsdale, Hixson Crowell, Weibull, and Korsmeyer-Peppas. The similarities between two in vitro dissolution profiles were assessed by the difference factor (f1), the similarity factor (f2) and the Rescigno index (xi(i)). The in vitro release kinetics of diltiazem hydrochloride tablets were evaluated using USP apparatus 4. Using a one way ANOVA (a = 0.05), statistically significant differences were found for t10%, t25%, and t50% dissolution times with a constant and with a variable pH dissolution fluid. The variation coefficient for the divisibility assay (Portuguese Pharmacopoeia VI) was lower than the limit value of 10%. The diltiazem release rate from this pharmaceutical system was not constant, and diminished with the square root of time (Higuchi model) showing that the phenomenon controlling drug release was the diffusion occurring inside the swelled polymeric matrix. Diltiazem release rate was a function of the area in direct contact with the dissolution fluid and not of the pharmaceutical matrix volume. The results obtained permit us to conclude that the division, in this case, affects the drug release characteristics. PMID- 11485177 TI - Corn starches as film formers in aqueous-based film coating. AB - The purpose of this study was to evaluate the film formation ability and mechanical stress-strain properties of aqueous native corn starches, using free films and film coatings applied to tablets. Free films were prepared from high amylose corn (Hylon VII), corn and waxy corn starches, using sorbitol and glycerol as plasticizers. The tablets and pellets were film-coated using an air suspension coater, and characterized with respect to the film coating surface topography, cross-sectional structure and thickness (SEM), and dissolution in vitro. The amylose content of the starch film formers affected both the tensile strength and the elongation. The elongations were under 5% for even the plasticized starches, and in most cases, no plasticization effect was seen by either of the plasticizers. Dissolution of native corn starch film-coated tablets (weight gain 1%) did not differ from uncoated ones. A notable delay in dissolution of the drug was found by increasing Hylon VII film coating thickness, suggesting controlled-release characteristics. PMID- 11485178 TI - An in vitro pulmonary permeation system with simulation of respiratory dynamics. AB - To study the effect of respiration on transpulmonary permeation kinetics of drugs, an in vitro pulmonary permeation system, which consists of a setup for the simulation of respiratory dynamics, was developed. The system is composed offour major components: a pair of horizontal-type half-cells, a model air-blood barrier, an instrument for the application and regulation of respiratory pressure, and a pressure monitoring system. Calibration studies were performed and results showed that the primary respiration parameters (the peak inspiration pressure, respiratory frequency, and the percent inspiration time) can be controlled at a reproducible manner. This system appears to simulate very well the respiratory dynamics observed normally under physiologic conditions. After calibration, the system was utilized to characterize and quantitate the effect of respiration on the transpulmonary permeation of drugs using progesterone as the model drug. The results showed that progesterone permeability is increased as much as 1.8-5.6 folds by application of a respiratory pressure, depending on the combination of respiration parameters. Further studies demonstrated that the enhancement in pulmonary permeation triggered by respiratory pressure is resulted from the stretching of the lung tissue, not by the pressure gradient itself. The observations lead to the conclusion that the system developed in this investigation is a useful in vitro tool for studying the kinetics of pulmonary drug permeation under a physiologically simulating respiratory dynamics. The studies have provided scientific evidence for demonstrating that respiration is an important factor in determining the kinetics of transpulmonary drug permeation through possible alteration in the properties of the air-blood barrier. PMID- 11485179 TI - Release of hydrocortisone from a cream matrix: dependency of release on suspension concentration and measurement of solubility and diffusivity. AB - The principal object of the present research was to investigate the sensitivity of drug release from a semisolid system to the manner of its preparation and to the concentration of drug placed within it. Established theory indicated that release should be concentration dependent, with the specific dependency determined by whether the drug in the system is fully in solution or is present substantially as suspended matter. To purposefully explore these relatively untested performance expectations, the total amount of drug in the formula was varied from a low concentration of 0.25% to a high concentration of 3%. Conditions were established such that drug release conformed to release from a semi-infinite medium into a receptor sink. At every concentration, release profiles were well reproduced in replicate samples and, where multiple lots of a kind were employed, from lot to identical lot. In all cases square root of time release kinetics were observed. Moreover and without exception, the square root of time release rate from run to run was directly proportional to the square root of the total concentration of hydrocortisone placed in the formulations. The amount released per square root of time per square root of total concentration was nearly identical from run to run irrespective of total concentration. The overall behavior fit theoretical expectations for suspensions having only a small fraction of the drug they contain in solution. That this condition prevailed even at the lowest 0.25% hydrocortisone strength was proven by independently measuring hydrocortisone's solubility (0.02%). Measurement of solubility permitted estimation of the effective diffusivity of the drug through the cream (2 x 10(-7) cm2/s). PMID- 11485180 TI - Multiunit controlled-release diclofenac sodium capsules using complex of chitosan with sodium alginate or pectin. AB - This study explored the application of chitosan-alginate (CA) and chitosan-pectin (CP) complex films as drug release regulator for the preparation of multiunit controlled-release diclofenac sodium capsules. Pellets containing drug and microcrystalline cellulose, in a ratio of 3:5, were prepared in a fluidized rotary granulator. The pellets were coated with CA, CP, sodium alginate, pectin, and chitosan solutions. The pellets, equivalent to 75 mg drug, were filled into capsules. After 2 h of dissolution test in acidic medium, the amount of the drug released from any preparation was negligible. The pellets were further subject to pH 6.8 phosphate buffer More than 80% drug release at 12 h was observed with the uncoated pellets and those coated with sodium alginate, pectin or chitosan. Both 1% CA and 3% CP coated pellets exhibited drug release profiles similar to that of Voltaren SR75. It was found that approximately 60% and 85% of the drug were released at 12 and 24 h, respectively. Both Differential thermal analysis (DTA) and Fourier transform infrared spectroscopy (FTIR) analyses revealed complex formation between chitosan and these anionic polymers. It could be concluded that CA and CP complex film could be easily applied to diclofenac sodium pellets to control the release of the drug. PMID- 11485181 TI - Glass vials for small volume parenterals: influence of drug and manufacturing processes on glass delamination. AB - PURPOSE: Studies were initiated to examine the effect of formulation and process variables on the delamination process and also the influence of the glass manufacturing process, supplier, and glass surface treatment. METHODS: Stress testing was performed by exposing filled vials to multiple sterilization cycles followed by accelerated stability testing. Delamination incidence was determined by visual examination, light obscuration (HIAC), and microscopical methods. The inner surface of vials from each supplier and lot were also examined by scanning electron microscopy. RESULTS: Vials sourced from Supplier A had smooth surfaces as demonstrated by SEM examination, whereas vials sourced from Suppliers B and C displayed extensive surface imperfections such as pitting and/or deposits. These imperfections were localized to the vial wall, adjacent to the vial bottom, and increased with sulfate treatment. Delamination incidence increased in those vial lots with increased surface imperfections. Thus, vials sourced from Supplier A had the lowest frequency of delamination. Sulfate treatment and high pH increased delamination incidence to as high as 100%. CONCLUSION: These results demonstrate the importance of the surface morphology created during the vial forming process. Given the diferences observed, final vial selection should include extensive microscopical and product stress testing studies on multiple vial lots. PMID- 11485182 TI - Evaluation of bioadhesive glipizide spheres and compacts from spheres prepared by extruder/marumerizer technique. AB - The objective of this study was to attempt to deliver glipizide from spheres and compacts containing the natural polymer Carrageenan (Gelcarin, GP 812) and prepared by extruder/marumerizer technique. A second objective was to evaluate the mucoadhesive strength of the bioadhesive spheres onto the mucus membrane of rabbit. The effects of polymer, drug level, and type of spheronizing material were evaluated. All sphere formulations were compacted into tablets using a rotary Manesty B-3B machine equipped with 12/32 flat face tooling. Results show drug release from spheres and compacts decreased as the level of Carrageenan was increased. However as the level of drug was increased drug release also increased. Spheres containing Avicel PH-101 gave higher drug release than spheres of the same composition but prepared with Avicel RC-581. In general, the drug release from tablets was higher than its corresponding spheres and drug release from spheres and tablets containing Carrageenan was higher than control spheres and tablets of the same composition but without Carrageenan. Tablet formulations compacted from spheres containing Avicel RC-581 gave higher release rate constants than tablet formulations of the same composition but prepared with Avicel PH-101. The bioadhesion study showed that mucoadhesion strength between spheres and mucus membrane of the rabbit depends on the levels of polymer, drug, and type of spheronizing material. Developed bioadhesive spheres and tablets increase the solubility of glipizide which may increase its bioavailability and also increased the adherence of the bioadhesive systems to the mucous membrane so that once daily dose can be administered. PMID- 11485183 TI - Evaluation of the functional equivalence of crospovidone NF from different sources. II. Standard performance test. AB - Current NF monographs do not provide tests that reflect on the functionality of Crospovidone NF from multiple sources. Physical characterization studies such as particle size and distribution, surface area, porosity, and surface morphology revealed major differences among the crospovidones from different sources (Shah, U.; Augsburger, L.L. J. Pharm. Dev. Technol. 2001, 6 (1), 39-51). Differences in disintegration and dissolution were also observed for a model drug in an insoluble filler system (see Shah and Augsburger, 2001). The objective of this study was to determine the relationship between physical differences observed and disintegrant functionality and to develop standard performance test. Tests performed included settling volume studies, measurement of initial rate as well as extent of liquid uptake of the loose disintegrant powder, and simultaneous measurement of the axial and radial disintegrating forces along with the rate and extent of liquid uptake of the pure disintegrant compacts. Significant differences among the crospovidones were observed for all tests performed. Settling volume, liquid uptake, and disintegration force are recommended as standard performance tests to determine differences among crospovidones from different sources. PMID- 11485184 TI - Sorption of parabens by flexible tubings. AB - Flexible tubings are extensively used in pharmaceuticals, food industry, and in hospitals. This study was undertaken to compare various flexible tubings to determine their sorption characteristics, using methyl and propyl parabens. After 24 h, some tubings showed 100% sorption of propylparaben and over 40% for methylparaben. Significant losses were observed within a few hours using several tubings. For methylparaben, the losses were in the following decreasing order of sorption: Tygon, Clearflo, silicone, Nylotube, and Newtex. For propylparaben, the losses were in the following order: Tygon, Clearflo, silicone, Newtex, and Nylotube. Teflon, Zelite, and Vitube showed little to no losses of methyl and propylparaben over 120 h of study. The silicone tubing, refilled after 120 h with fresh methylparaben or propylparaben solutions, again showed significant losses within a few hours. The tubings show slow desorption when filled with the buffer vehicle. For Silastic tubing, increase in temperature from 25 to 40 degrees C, increase in pH from 3.5 to 6.5, tubing lot to lot variation, or curing with peroxide or platinum had little or no effect on paraben sorption. As expected, the sorption of parabens increased with increasing surface area of Silastic tubing. Results provided can be used to select the best tubings and to minimize paraben losses during production and filling of liquid pharmaceuticals andfood products containing these antimicrobial preservatives. PMID- 11485185 TI - Sustained in vivo activity of recombinant bovine granulocyte colony stimulating factor (rbG-CSF) using HEPES buffer. AB - The purpose of this study was to develop a long-acting injectable formulation of bG-CSF for veterinary use. However, in order to achieve sustained in vivo activity it was first necessary to stabilize the protein at the injection site. Preformulation studies, as well as literature, suggest that bG-CSF aggregates at neutral pH ranges (i.e., pH 6-8) and at temperatures of approximately 40 degrees C. Therefore, bG-CSF will not retain its activity for an extended period of time at the injection site. During this study we determined that HEPES buffer has a very significant impact on protein stability as well as on biological performance. Recombinant bovine granulocyte colony stimulating factor (rbG-CSF) was formulated in 1 M HEPES buffer for subcutaneous injection into cows. bG-CSF formulated in 1 M HEPES buffer resulted in sustained in vivo activity of bG-CSF compared to the "control" formulation (control formulation: 5% mannitol, 10 mM acetate buffer, 0.004% tween-80, pH 4). White blood cell (WBC) count was used as a marker to evaluate in vivo activity of the formulation. WBC numbers remained above a threshold value for only 24-30 h for the control formula. However, when bG-CSF was formulated in 1 M HEPES, the WBC remained above threshold for 3 days or 72 h. Formulating bG-CSF in 1 M HEPES at pH 7.5 also resulted in greater solution stability. This was surprising since bG-CSF is intrinsically not stable at neutral pH. The effect of 1 M HEPES on the T(M) (temperature at maximum heat flow on calorimetry scan) of bG-CSF was determined by microcalorimetry. In the absence of 1 M HEPES buffer the T(M) was 48 degrees C (onset approximately 40 degrees C), while bG-CSF formulated in 1 M HEPES buffer has a T(M) of 59 degrees C (onset approximately 50 degrees C). Similar organic buffers, such as MOPS, HEPPS, TES, and tricine, also resulted in improved solution stability as well as in sustained in vivo activity. The dramatic effect of these buffers on stability and biological performance of bG-CSF is not well understood. One hypothesis is that the electrostatic interaction between the zwitterionic form of these buffers and bG-CSF provides stabilization against denaturation. PMID- 11485186 TI - Preparation and characterization of spray-dried oxidized cellulose microparticles. AB - The goal of this study was to investigate the feasibility of spray drying to produce microparticles of oxidized cellulose (OC), a biocompatible and bioresorbable polymer. OCs containing 7, 13, and 20 wt% carboxylic groups were converted into stable aqueous dispersions and then spray dried using a Yamoto G 32 spray dryer equipped with a standard fluid nozzle with an orifice of 406 microm. The following operating conditions were investigated: inlet temperature 140, 170, and 190 degrees C; feed rate 3, 6, and 9 mL/min; and atomization airpressure 0.5, 1, and 1.5 kg f/cm2. The amounts of OC used in feed were 1, 2.5, and 5%. OC microparticles produced under these conditions were shrunken spheres, ranging in size between 0.98+/-0.47 and 2.05+/-0.98 microm. The different operating conditions used had no significant effect on the size and shape of particles. The use of a water-soluble plasticizer (glycerin, polyethylene glycol 400, or polyethylene glycol 6000) in the dispersion yielded microparticles with a good sphericity and a smooth surface morphology, whereas no change in the shape or size of microparticles was noted with water-insoluble plasticizers, Triacetin and dibutyl phthalate. Powder X-ray diffraction and Fourier transform infrared spectral analyses of spray-dried microparticles showed no change in the solid state structure of OC. In conclusion, results show that OC can be converted into stable aqueous dispersions and used to produce microparticles by spray drying. PMID- 11485187 TI - Improved dissolution and cytotoxicity of camptothecin incorporated into oxidized cellulose microspheres prepared by spray drying. AB - Oxidized celluloses (OC) containing 7, 13, and 20% carboxylic content (OC-7, OC 13, and OC-20, respectively) have been converted into aqueous colloidal dispersions and used to prepare microspheres of the antineoplastic agent camptothecin (CPT) by spray drying. Plasticizers used were glycerin, polyethylene glycol 400 (PEG-400), and polyethylene glycol 6000 (PEG-6000). Irrespective of the carboxyiic content of OC and the nature of plasticizer employed, the size of microspheres varied from 1.25+/-0.40 to 1.52+/-0.47 microm. The release studies in pH 7.4 buffer revealed the dissolution of CPT to be faster from the microsphere formulations than from physical mixtures and free CPT. The times to release 50% CPT (T-50%) from microspheres prepared using OC-7, OC-13, and OC-20 were about 31, 37, and 19 h, respectively. The in vitro cytotoxicity results indicated OC-20/CPT microspheres to be more effective than free CPT against human derived RPMI-8402 lymphoid and THP-1 myeloid leukemia cell lines. The ED50 values for the OC-20/CPT microspheres and free CPT were 1 x 10(-5) and 0.25 x 10(-1) microg/mL, respectively, against the RPMI-8402 line and 0.5 x 10(-2) and 0.75 microg/mL, respectively, against the THP-1 line. The higher activity of OC-20/CPT microspheres compared to that of the free drug is attributed to increased dissolution of CPT from microspheres. PMID- 11485189 TI - Effective graphical displays. AB - A graph is a visual display of data to achieve an understanding of the underlying patterns and interrelationships of the data. Such visual representations of data often form the basis for inferences and decisions. However, a poorly chosen graphical form can lead to erroneous inferences. In this paper previously published graphs depicting biopharmaceutical, drug metabolism, formulation, and pharmacokinetic data are recast using alternate display methods in an attempt to better clarify the data structure. The display methods used include dot plots and trellis plots. PMID- 11485188 TI - Investigation of the utility of an in vitro release test for optimizing semisolid dosage forms. AB - Current literature indicates that an in vitro release test (IVRT) can serve as a research tool during the course of developing topical formulations. The purpose of this study was therefore to investigate the ability of an IVRT to select the topical semisolid formulations with the most rapid release rate of the model drug ketoprofen from two closely related hydrogels in a simulated product development process. Two glycols with distinct differences in their physical-chemical properties, Transcutol P (ethoxydiglycol) and propylene glycol, were incorporated into Carbopol 980 and Poloxamer 407 formulations. The release rate of ketoprofen was determined utilizing different receptor media and conditions, i.e., phosphate buffer pH 7.4, isopropyl myristate (IPM), and a combination of an IPM soaked membrane and phosphate buffer (pH 7.4) as receptor fluid. The results indicated that the conditions chosen could affect greatly the conclusions concerning the formulations. The only observable trend was that Transcutol P-containing formulations tended to permit a faster ketoprofen release than propylene glycol containing formulations when utilizing IPM as a receptor component. This was attributed to the mutual miscibility of Transcutol P in IPM. It can be concluded that, for the purpose of formulation screening in the early phases of product development, an IVRT will only be useful for predicting the amount of drug available for absorption if the receptor medium has properties that closely mimic human skin. These results illustrate the importance of selecting suitable receptor components and indicate that it may be necessary to consider alternatives to the commonly used synthetic membranes. PMID- 11485190 TI - Spatial attention in early vision. AB - The present study addressed whether the allocation of attention to a particular region in space can prevent processing of distractor information from non attended regions. A cue indicated the area in visual space where the target singleton would be presented. Observers were required to detect this target singleton and ignore a distractor singleton presented within a non-attended region. The results indicate that the allocation of attention to a region in space cannot prevent the processing of unwanted information from elsewhere in the visual field. It is concluded that the function of the allocation of attention is not to enhance the processing capacity within the attended region but rather to attenuate interference from distractors in unattended regions. PMID- 11485191 TI - Directing spatial attention towards the illusory location of a ventriloquized sound. AB - In this study, we examined whether ventriloquism can rearrange external space on which spatial reflexive attention operates. The task was to judge the elevation (up vs down) of auditory targets delivered in the left or the right periphery, taking no account of side of presentation. Targets were preceded by either auditory, visual, or audiovisual cues to that side. Auditory, but not visual cues had an effect on the speed of auditory target discrimination. On the other hand, a ventriloquized cue, consisting of a tone in central location synchronized with a light flash in the periphery, facilitated responses to targets appearing on the same side as the flash. That effect presumably resulted from the attraction of the apparent location of the tone towards the flash, a well-known manifestation of ventriloquism. Ventriloquism thus can reorganize space in which reflexive attention operates. PMID- 11485192 TI - Attention and size in a global/local task. AB - Two-letter stimuli, consisting of one small letter inside a much larger one (in Experiments 1A, 1B, and 2) or inside a "blob" (in Experiment 3), were used to examine the role of size difference in global/local tasks. The small letter was placed at locations that avoided contour interactions. The results showed no identity interference, in that the specific identity of the large letter did not differentially affect identification of the small one. However, there was evidence of global advantage, in that the presence of a large letter hindered identification of the small one. The magnitude of the global advantage effect, as measured by the difference in performance between the small-single and small embedded conditions, was largest (about 200 ms reaction time (RT) difference) when the large letters were the same as the small ones, lower (a 63 ms difference in Experiment IB, and 89 ms in Experiment 2) when the large letters were unrelated to the small ones, and lowest (a 25 ms difference) when the large stimuli were blobs. It is proposed that the amount of interference depends on the overlap between the features of the large stimuli, as a set, and those of the small ones, also as a set. PMID- 11485193 TI - The one-of-us effect in decision evaluation. AB - Judgment-by-outcomes denotes basing retrospective evaluation of decisions on the valence of their outcomes (success versus failure). Although decisions are typically evaluated in social contexts, so far judgment-by-outcomes has been studied without regard to this context. This study examines the moderating effect of evaluator's identification with the decision maker (the one-of-us-effect) on the influence of outcome information on the evaluation of Arab and Jewish subjects were presented with two cases recounting operations by either Arab or Jewish underground directed against the British authorities in Palestine. One case was a success (from the underground's point of view) and one ended in failure. Consistent with the one-of-us effect, identification with the decision maker variably canceled the influence of outcome information altogether, accentuated or weakened its influence, or determined which outcome constituted successful and unsuccessful outcomes. The one-of-us effect exercised a differential influence over different facets of decision evaluation, influencing most strongly the assignment of sanctions (in-group decision makers were mostly rewarded, out-group decision-makers were mostly punished regardless of outcomes). Next, in order of potency, the effect influenced the evaluation of decision justification, the evaluation of the decision maker, and the evaluation of the quality of decision process. PMID- 11485194 TI - Negative priming in a gender decision task and in a semantic categorization task. AB - In three experiments, we tested the hypothesis that negative priming (NP) can occur without prime or target selection, when conflicting properties are associated with the prime and the target, and when the experimental conditions allow the encoding of the target as a separate episode from the prime. These predictions were confirmed in Experiment 1, using a gender decision task. Responses were slower when prime and target had the same gender than when they had different genders, with an inter-stimulus interval (ISI) of 600 ms but not with an ISI of 25 ms. Experiment 2 eliminated a possible explanation of the NP obtained in Experiment 1, in terms of response inhibition during the prime processing. Finally, Experiment 3 demonstrated the replicability and the generality of our NP, in a semantic categorization task. Empirical and theoretical consequences of our results for studies using the priming paradigm are discussed. PMID- 11485195 TI - Up-regulation of genes encoding novel extracellular proteins during fruit set in pea. AB - The transition from the carpel of the flower to a developing fruit is a poorly characterized process despite its agricultural importance. We have identified two genes, GIC19 and GIC4, which are expressed after induction of pea (Pisum sativum L.) fruit set either by exogenous gibberellins or by pollination. GIC19 expression is temporally and spatially regulated, with transcripts mainly found in growing carpels and young fruit. Similar to GIC19, GIC4 expression is developmentally regulated during carpel and fruit development. However, GIC4 transcripts are found in other growing tissues throughout the plant. Analysis of their sequences and localization of fusion proteins with GFP indicate that both GIC19 and GIC4 are extracellular proteins. While GIC19 is a small proline-rich protein with no overall homology to other reported proteins. GIC4 belongs to a novel family of proteins. Our results reinforce a model of gibberellin mode of action during pea fruit set and development involving enhanced synthesis of extracellular proteins and secretory activity to provide materials and energy for cell growth. PMID- 11485196 TI - The poplar bark storage protein gene (Bspa) promoter is responsive to photoperiod and nitrogen in transgenic poplar and active in floral tissues, immature seeds and germinating seeds of transgenic tobacco. AB - In Populus, seasonal nitrogen storage involves the accumulation of a 32 kDa bark storage protein (BSP) in the inner bark parenchyma and xylem rays. Poplar BSPs are encoded by a multigene family and one member, bspA, has been cloned and sequenced. The regulation of bspA was investigated by transforming either hybrid poplar or tobacco with a chimeric gene consisting of the 2.8 kb bspA promoter fused to the coding region of beta-glucuronidase (uidA). In transformed poplar, the bspA 2.8 kb promoter conferred both short-day (SD) and nitrogen (N) inducibility to GUS and activity was localized to the bark (primary and secondary phloem, and cortex) and xylem rays. Night-break treatments inhibited SD induction of GUS. Deletion of the 1.6 kb distal DNA sequences from the bspA promoter eliminated SD induction of GUS while some N induction was retained. These results indicate that although poplar BSP is encoded by a multigene family, transcriptional activation of bspA per se can account for bsp expression in bark and xylem rays in response to either SD or N treatment. These results also show that the elements responsible for SD or N induction are separable. Because of the long generation intervals associated with trees, the developmental regulation of bspA in flowers, developing seeds, and germinating seeds was investigated by transforming the 2.8 kb bspA-promoter::uidA chimeric gene into tobacco. The bspA promoter was active in developing tobacco floral tissues and in seeds during early stages of embryogenesis, decreased progressively during seed maturation and regained activity upon seed germination. Although seed storage proteins of poplar share some similarities to poplar BSP, the observed developmental expression patterns in tobacco are consistent with a role for bspA in vegetative rather than seed storage protein storage. PMID- 11485197 TI - The glutathione peroxidase homologous gene from Chlamydomonas reinhardtii is transcriptionally up-regulated by singlet oxygen. AB - The glutathione peroxidase homologous gene (Gpxh gene) in Chlamydomonas reinhardtii is up-regulated under oxidative stress conditions. The Gpxh gene showed a remarkably strong and fast induction by the singlet oxygen-generating photosensitizers neutral red, methylene blue and rose Bengal. The Gpxh mRNA levels strongly increased, albeit much more slowly, upon exposure to the organic hydroperoxides tert-butyl hydroperoxide (t-BOOH) and cumene hydroperoxide. In contrast, the Gpxh mRNA levels were only weakly induced by exposure to the superoxide-generating compound paraquat and by hydrogen peroxide. A comparison of the Gpxh mRNA levels with those of the heat shock protein HSP70A and the iron superoxide dismutase gene showed qualitative and quantitative differences for the three genes under oxidative stress conditions tested. The Gpxh gene is specifically induced by singlet-oxygen photosensitizers and the relative induction by other compounds is much weaker for Gpxh than for the other genes investigated. Using Gpxh promoter fusions with the arylsulfatase reporter gene, we have shown that the Gpxh was transcriptionally up-regulated by singlet-oxygen photosensitizers. It is also shown that the Gpxh promoter contains a region between 104 and 179 bp upstream of the transcription start that is responsible for the mRNA up-regulation upon exposure to 1O2 but not t-BOOH. Within this region a regulatory sequence homologous to the mammalian cAMP response element (CRE) and activator protein 1 (AP-1) binding site was identified within a 16 bp palindrome. PMID- 11485198 TI - Alternative splicing of prosystemin pre-mRNA produces two isoforms that are active as signals in the wound response pathway. AB - Systemin and its precursor protein, prosystemin, play an essential role in the systemic wound response pathway of tomato plants. We report here the isolation from tomato of a novel prosystemin cDNA (prosysB) that differs from the reported cDNA sequence (prosysA) by the addition of a CAG trinucleotide. Inspection of the prosystemin genomic sequence, which was mapped to the central region of chromosome 5, indicated that prosysA and prosysB transcripts are generated by an alternative splicing event that utilizes different 3' splice sites within intron 3. Quantitative RT-PCR analysis showed that prosysB transcripts accumulated to approximately twice the level of prosysA in all tissues that express the prosystemin gene. The relative abundance of the two mRNAs was unaffected by wounding or methyl jasmonate treatment, conditions that increase the level of total prosys mRNA. These findings indicate that alternative splicing of prosys pre-mRNA is a constitutive process. The amino acid sequence of prosysB is predicted to differ from that of prosysA by replacement of Arg-57 with Thr-Gly in the non-systemin portion of the protein. Over-expression of the prosysB cDNA in transgenic tomato plants conferred constitutive expression of defense genes that are regulated by wounding and systemin. We conclude that prosysB is the major prosystemin-encoding transcript in tomato, and that this isoform is active as a signal in the wound response pathway. PMID- 11485199 TI - Functional expression and subcellular localization of the Nectria haematococca Mak1 phytoalexin detoxification enzyme in transgenic tobacco. AB - Medicarpin and maackiain are antifungal pterocarpan phytoalexins produced by many legumes, and are thought to be important components of the defense response of these legumes to certain fungal pathogens. The Mak1 gene from the fungal pathogen Nectria haematococca encodes an FAD-dependent mono-oxygenase, known to specifically hydroxylate the phytoalexins medicarpin and maackiain, converting them to less fungitoxic derivatives. Two binary vector constructs were made containing the coding regions from two fungal clones, a Mak1 cDNA (intronless) and a genomic (including three fungal introns) clone, regulated by an enhanced cauliflower mosaic virus 35S promoter. The constructs were introduced into tobacco to check for expression of active fungal enzyme in plant cells and for splicing of fungal introns. Leaves of tobacco plants transformed with the Mak1 cDNA construct readily metabolized infiltrated medicarpin to 1a hydroxymedicarpin, indicating high levels of active enzyme. RT-PCR analysis of tobacco plants transformed with the Mak1 genomic construct indicated no processing of Mak1 introns, and no Mak1 activity was detected in these plants. When using plants containing the Mak1 cDNA construct, immunolocalization with a Mak1-specific antibody together with cellular fractionation indicated that Mak1 protein accumulated in the plant cytoplasm, associated with endoplasmic reticulum membranes; medicarpin biosynthetic enzymes have been localized to the same subcellular region. The Mak1 cDNA construct is therefore suitable for use in studies to selectively eliminate medicarpin accumulation to assess the relative importance of medicarpin in the antifungal defense mechanisms of alfalfa and other legumes. PMID- 11485200 TI - Transgene silencing of invertedly repeated transgenes is released upon deletion of one of the transgenes involved. AB - To analyse experimentally the correlation between transgene silencing and the presence of an inverted repeat in transgenic Arabidopsis thaliana plants, expression of the beta-glucuronidase (gus) gene was studied when present as a convergently transcribed inverted repeat or as a single copy in otherwise isogenic lines. In transformants containing two invertedly repeated gus genes separated by a 732 bp palindromic sequence, gus expression was low, as exemplified by the expression levels in the parental line KH15. The parental KH15 locus could induce efficiently in trans silencing of gus copies at allelic and non-allelic positions. In transformants containing two invertedly repeated gus genes separated by a 826 bp non-repetitive spacer region, gus expression was high or intermediate, especially in hemizygous state and at late developmental stages, as demonstrated in detail for line KHsb67. Removal of one of the gus copies by Cre recombinase resulted in all cases in constitutively high gus expression in hemizygous as well as in homozygous state. The derived deletion lines could no longer induce in trans silencing of homologous gus copies. The results show that convergent transcription of transgenes in an inverted repeat is an important parameter to trigger their silencing and that co-transformation of two T-DNAs with identical transgenes can be used to obtain inverted repeats and targeted co suppression of the homologous endogenes. Moreover, the data suggest that the spacer region in between the inverted genes plays a role in the efficiency of initiating and maintaining silencing. PMID- 11485201 TI - One of two tandem Arabidopsis genes homologous to monosaccharide transporters is senescence-associated. AB - A gene designated SFP1, which is similar to major facilitator superfamily monosaccharide transporters, is induced during leaf senescence. Genomic sequence analysis identified a second highly similar and closely linked gene, SFP2, suggesting that SFP1 and SFP2 may have arisen through a recent duplication event. However, RNA gel-blot analyses and histochemical localization of a reporter gene activity in transgenic plants show that SFP1 and SFP2 are differentially regulated and that only SFP1 is induced during leaf senescence. The increase in SFP1 gene expression during leaf senescence is paralleled by an accumulation of monosaccharides. Possible roles for SFP1 in sugar transport during leaf senescence are discussed. PMID- 11485202 TI - A tomato homologue of the human protein PIRIN is induced during programmed cell death. AB - Programmed cell death (PCD), with similarities to animal apoptosis, was induced in tomato suspension cells by the topoisomerase I inhibitor camptothecin. Previously, a differential display screening was performed to isolate genes differentially expressed during camptothecin-induced cell death. As a result, the new tomato gene Le-pirin was isolated, whose mRNA levels dramatically increase during camptothecin-induced PCD. Le-pirin mRNA accumulation is also observed when cell death is triggered by the mycotoxin fumonisin-B1, but not when the suspension cells are treated with stress-related compounds such as ethylene, methyl jasmonate or salicylic acid. The caspase inhibitor Z-Asp-CH2-DCB and the calcium channel blocker LaCl3 effectively delayed whereas ethylene greatly stimulated camptothecin-induced PCD and the accumulation of Le-pirin mRNA. The Le pirin encoded protein shows 56% identity with the human protein PIRIN, a nuclear factor reported to interact with the human oncogene Bcl-3. Human PIRIN stabilizes the formation of quaternary complexes between Bcl-3, the anti-apoptotic transcription factor NF-kappaB and its DNA target sequences in vitro. The isolation of Le-pirin and its implication in plant PCD provides new clues on the role of putative NF-kappaB-associated pathways in plant defence mechanisms. PMID- 11485203 TI - Analysis of a dehiscence zone endo-polygalacturonase in oilseed rape (Brassica napus) and Arabidopsis thaliana: evidence for roles in cell separation in dehiscence and abscission zones, and in stylar tissues during pollen tube growth. AB - The oilseed rape (Brassica napus) endo-polygalacturonase (endo-PG) RDPG1 is involved in middle lamella breakdown during silique opening. We investigated tissue-specific expression of RDPG1 in transgenic Arabidopsis thaliana. Cellular localization of endo-PG protein in Arabidopsis siliques was determined by immuno electron microscopy. An Arabidopsis orthologue, ADPG1, was isolated and aligned with the sequence of RDPG1. The proximal 5' sequences as well as introns are largely conserved. Analysis of the histological GUS-staining pattern of two RDPG1 promoter-GUS (beta-glucuronidase) constructs in transgenic Arabidopsis revealed that the conserved proximal part of the 5'-flanking region directs expression in dehiscence zones of siliques and anthers, floral abscission zones and stylar tissues during pollen tube growth, branch points between stems and pedicel and expression associated with the apical meristem of seedlings, while the distal part of the RDPG1 5'-flanking region contains elements involved in vascular associated expression in petals, cotyledons and roots. Subsequent RT-PCR analysis, on RNA from the corresponding rape tissues, confirms the staining pattern revealed in transgenic Arabidopsis, thereby justifying the use of Arabidopsis as a reliable model system for analysis of oilseed rape regulatory sequences. PMID- 11485204 TI - The protease and reverse transcriptase of the tobacco LTR retrotransposon Tnt1 are enzymatically active when expressed in Escherichia coli. AB - The open reading frame (ORF) of the tobacco retrotransposon Tnt1-94 was over expressed in Escherichia coli to assay its protease and reverse transcriptase (RT) enzymatic activities. In E. coli, Tnt1-94 polyprotein is cleaved off by the element-encoded protease to release a Gag protein with an apparent molecular mass of 37 kDa that forms high-density aggregates. The catalytic site of Tnt1-94 protease (D-T-A) as determined by deletion analysis differs from that of retroviruses and of well-characterized retrotransposons (D-T/S-G). The cleaved or uncleaved ORF of Tnt1-94 displays an exogenous RT activity. Over-expression of plant retrotransposons ORFs in E. coli provides a very useful strategy to assay the enzymatic activities of their proteins and to determine their catalytic sites. PMID- 11485205 TI - Differential expression of alpha- and beta-expansin genes in the elongating leaf of Festuca pratensis. AB - Grasses contain a number of genes encoding both alpha- and beta-expansins. These cell wall proteins are predicted to play a role in cell wall modifications, particularly during tissue elongation. We report here on the characterisation of five alpha- and three vegetative beta-expansins expressed in the leaf elongation zone (LEZ) of the forage grass, Festuca pratensis Huds. The expression of the predominant alpha-expansin (FpExp2) was localised to the vascular tissue, as was the beta-expansin FpExpB3. Expression of another beta-expansin (FpExpB2) was not localised to vascular tissue but was highly expressed in roots and initiating tillers. This is the first description of vegetative beta-expansin gene expression at the organ and tissue level and also the first evidence of differential expression between members of this gene family. In addition, an analysis of both alpha- and beta-expansin expression along the LEZ revealed no correlation with growth rate distribution, whereas we were able to identify a novel xyloglucan endotransglycosylase (FpXET1) whose expression profile closely mimicked leaf growth rate. These data suggest that alpha- and beta-expansin activities in the grass leaf are associated with tissue differentiation, that expansins involved in leaf growth may represent more minor components of the spectrum of expansin genes expressed in this tissue, and that XETs may be useful markers for the analysis of grass leaf growth. PMID- 11485206 TI - Type I interferons and herpes simplex virus infection: a naked DNA approach as a therapeutic option? AB - Herpes simplex virus (HSV) types 1 and 2 are highly successful human pathogens that can elicit blinding herpetic keratoconjunctivitis, fatal sporadic encephalitis, aseptic meningitis, and increase the risk of acquiring additional sexually transmitted diseases. Type I interferons (IFN) play a significant role in controlling HSV pathogenesis by antagonizing viral replication and spread. Taking advantage of the susceptibility of HSV to IFNs, a novel approach of employing plasmid DNA cassettes expressing type 1 IFNs to antagonize viral pathogenesis has been undertaken. This review will describe recent work in our lab and those of others using naked DNA encoding cytokines to antagonize HSV replication and virus trafficking or immune-mediated pathogenesis as a result of viral assault to ocular tissue. PMID- 11485207 TI - The HMG I proteins: dynamic roles in gene activation, development, and tumorigenesis. AB - The high mobility group I, Y, and I-C proteins are low-molecular-weight, nonhistone chromosomal proteins that play a general role modulating gene expression during development and the immune response. Consistent with their role in early development, all three proteins are expressed at high levels during embryogenesis, and their expression is markedly diminished in differentiated cells. Exceptions to the general repression of these genes in adult tissues involve (1) A burst of synthesis of the HMG I protein during the immune response (during lymphocyte activation and preceding cytokine/adhesion molecule gene expression), (2) A constitutive expression of the HMG I and Y proteins in photoreceptor cells, and (3) Derepression of HMG I, Y, and often I-C expression in neoplastic cells. Work from several laboratories has now uncovered how these proteins participate in gene activation: (1) By altering the chromatin structure around an inducible gene-and thus influencing accessibility of the locus to regulatory proteins-(2) By facilitating the loading of transcription factors onto the promoters, and (3) By bridging adjacent transcription factors on a promoter via protein/protein interactions. Despite the similar structures and biochemical properties of the three proteins, the work has also provided clues to a division of labor between these proteins. HMG I and Y have demonstrable roles in enhanceosome formation, whereas HMG I-C has a specific role in adipogenesis. C terminal truncations of HMG I-C and wild-type HMG Y appear to function in a manner analogous to oncogenes, as assessed by cellular transforation assays and transgenic mice. Future work should clearly define the similarities and differences in the biological roles of the three proteins, and should evolve to include attempts at pharmaceutical intervention in disease, based upon structural information concerning HMG I interactions with DNA and with regulatory proteins. PMID- 11485209 TI - Cellular immunity and memory to respiratory virus infections. AB - Respiratory virus infections, such as those caused by influenza and parainfluenza viruses, are a major cause of morbidity and mortality worldwide. Current vaccines against these pathogens rely on the induction of humoral immune responses that target viral coat proteins. Although this type of immunity provides solid protection against homologous virus strains, it is ineffective against heterologous virus strains that express serologically distinct coat proteins. In contrast, cellular immune responses can target internal antigens that are shared between heterologous viral strains. This form of immunity, sometimes referred to as heterosubtypic immunity, can mediate a substantial degree of protection. Thus, vaccines that emphasize cellular immune responses would be a valuable complement to available humoral vaccines. However, we only have a rudimentary understanding of which T cell subsets mediate protective immunity, how T cell memory is established and maintained, how that memory is recalled in a secondary infection, and why cellular immunity wanes rapidly with time. Here we review the role of CD4+ and CD8+ T cells in the recall response to influenza and parainfluenza viruses. In particular we focus on the recent observation that substantial numbers of memory T cells are established in the lung tissues and discuss the potential role of these cells in mediating a recall response. A thorough understanding of the cellular immune response to infection in the lungs is essential for future vaccine development. PMID- 11485210 TI - Regulatory role of CD4+ T cells during the development of contact hypersensitivity responses. AB - Contact hypersensitivity (CHS) is a T cell-mediated immune response to cutaneous sensitization and subsequent challenge with haptens such as dinitrofluorobenzene and oxazolone. Many aspects concerning the development and regulation of CHS remain unknown. Using CHS as a model of T cell-mediated immune responses to antigens deposited in the skin we have studied the development and function of effector and regulatory T cell components of this response. These studies have revealed the effector role of hapten-specific CD8+ T cells in this response. In contrast, hapten-specific CD4+ T cells negatively regulate the magnitude and duration of the response. In this article we propose a model in which the CD4+ T cell compartment regulates the development of effector CD8+ T cells during sensitization for CHS and discuss potential mechanisms that CD4+ T cells might utilize to mediate this regulation. PMID- 11485208 TI - T cell signal transduction and the role of CD7 in costimulation. AB - The complex cellular interactions that govern the mammalian immune response are now known to include specific receptor/ligand interactions, recruitment of intracellular signaling molecules, activation of both kinases and phosphatases, and redistribution of macromolecular complexes into specific subcellular membrane locations that, in aggregate, result in transcriptional activation. While the TCR CD3 signal is critical for activation of the resting T cell, it alone is not sufficient to initiate transcriptional activation or generate an effective immune response. A number of other coreceptor molecules, including CD4, CD8, and CD28, have now been characterized that also play important roles in initiating or amplifying the activation of the T cell. A 40 kDa member of the immunoglobulin superfamily, the CD7 molecule, has also been shown to have costimulatory activity and to induce tyrosine and lipid kinase activities. Here we will review the signaling pathways initiated by TCR, CD28, and CD7, as well as the functional consequences of signal transduction through these receptors. PMID- 11485211 TI - The complement system in central nervous system diseases. AB - The activation of complement system is an important factor participating in inflammatory, neurodegenerative, and cerebrovascular diseases. Astrocytes and neurons are able to synthesize complement components. Myelin and oligodendrocyte (OLG) activate the classical pathway of complement in vitro in the absence of antibodies. Sublytic C5b-9 in the absence of cell death induces proto-oncogenes, activates cell cycle, and enhances cell survival in OLG. In addition, C5b-9 reverses the differentiation phenotype in OLG and enhances cell survival. beta amyloid protein is an activator of the complement system and neurons are susceptible to bystander complement mediated damage. These findings indicate that complement activation and membrane assembly of C5b-9 play an important role in pathogenesis of central nervous system (CNS) disorders. PMID- 11485213 TI - Distribution and hydrodynamic model of the Keumdong oil spill in Kwangyang Bay, Korea. AB - Oil concentrations in the seawater, seabed sediments, and shoreline sediments were measured from the samples collected 165 days after the Keumdong oil spill accident, which had occurred in Kwangyang Bay of the South Sea, Korea in September 1993. Distribution patterns of the concentrations in the seabed and shoreline sediments have allowed us to hindcast the transport and fate of the spilled oil. Overall, the patterns agree better with the southward (seaward) surface water circulation than with the northward (landward) bottom water circulation over the region. Rapid, initial dispersal of the spilled oil to the south and its subsequent grounding onto the intertidal seabed sediments, as well as onto the shorelines, appear to have made ineffective the subsurface oil transport by the bottom current. PMID- 11485212 TI - Pathways and bottlenecks in the web of inflammatory adhesion molecules and chemoattractants. AB - Leukocyte recruitment requires capture, rolling, activation, adhesion, and transmigration. Adhesion molecules and chemoattractants have been identified that mediate each of these steps. Their functions often overlap, but the combined absence of some molecules can lead to severe spontaneous phenotypes, as seen in CD18-/- mice, or early lethality, as in CD18-/- E-selectin-/- mice. These groups of molecules define bottlenecks that restrict the inflammatory response. Adhesion molecules and activation mechanisms can also form groups of preferential usage, or pathways. Based on these findings, a web-like model may represent the inflammatory process better than the linear cascade model. Bottlenecks and pathways depend on the degree and nature of overlapping functions, the disease process, tissue site, and the inflammatory stimulus. PMID- 11485214 TI - Initial results of environmental monitoring in the Texas Rio Grande Valley. AB - Previous studies have suggested that a segment of human disease may be attributable to environmental exposures. These may include exposure to chemicals released from a broad range of natural and man-made sources. The purpose of this study was to develop the sampling methodology and prepare a preliminary database on the presence of various organic chemicals in environmental media in two South Texas counties bordered by the Rio Grande River. A third county, located approximately 150 miles north of the Rio Grande River, was also sampled. The South Texas counties were the focus of study due to an increased incidence of anencephalic births in recent years. The environmental media that was sampled included surface water and sediment from the Rio Grande River and irrigation canals, as well as soil from adjacent cropland and pastures. Samples were collected using United States Geological Survey (USGS) quadrangle maps (7.5'; 1:24,000 scale) to identify the area of interest. At least one sampling location was established in each quadrangle. A pond sampler was used for the collection of surface water samples, while soil was collected with a stainless steel trowel. Sediment samples were collected directly in a glass jar. Solid samples were extracted in a soxhlet extractor using methylene chloride. Organic chemicals were concentrated from water samples on a Sep-Pak cartridge and the organics eluted with methanol/acetonitrile. Extracts were analyzed using GC-MS. All of the surface water samples contained aliphatic hydrocarbons and plasticizers, while soil samples contained aliphatics, plasticizers, pesticides, and industrial estrogens. Specific chemicals detected in environmental samples included atrazine and benzene dicarboxylic acid. Contaminant levels in sediments were generally higher than were detected in other media. The results demonstrate the broad variability of contaminant types and concentrations in environmental samples. Although this study presents only a very preliminary characterization of a large area of South Texas, the data indicate a number of pesticides and xenobiotic estrogens that were identified in environmental samples. Additional data providing more details of spatial and temporal distribution of contaminants as well as wildlife studies are needed. PMID- 11485215 TI - Increased catecholamine levels in urine in subjects exposed to road traffic noise: the role of stress hormones in noise research. AB - The nocturnal excretion of catecholamines in urine was studied in 30-45-year-old women whose bedroom and/or living room were facing streets of varying traffic volume. The traffic volume of the streets was used as an indicator of noise exposure; adrenaline and noradrenaline concentrations were assessed as indicators of the outcome of the physiological stress. Significant associations between traffic volume and noradrenaline concentrations in urine were found with regard to the exposure of the bedroom (not the living room), indicating a higher chronic physiological arousal in noise-exposed subjects as compared to less exposed. Subjective measures of disturbance due to traffic noise were positively correlated with the noradrenaline level. However, this was only found in subjects where closing the window could not reduce the perceived disturbance, which points to the effectiveness of individual coping mechanisms. Stress hormones are useful indicators to study associations, mechanisms, and interactions between noise, health outcomes, and effect modifiers in epidemiological noise research. PMID- 11485216 TI - Impacts of atrazine in aquatic ecosystems. AB - A portion of all herbicides applied to forests, croplands, road sides, and gardens are inevitably lost to water bodies either directly through runoff or indirectly by leaching through groundwater into ephemeral streams and lakes. Once in the aquatic environment, herbicides may cause stress within aquatic communities and radically alter community structure. Atrazine is one of the most effective and inexpensive herbicides in the world and is consequently used more frequently than any other herbicide. Atrazine is frequently detected in aquatic waters, and has been known to affect reproduction of aquatic flora and fauna, which in turn impacts on the community structure as a whole. This paper presents a summary of the reported direct and indirect impacts of atrazine on aquatic organisms and community structure. The information can be used for developing improved management guidelines and legislation. It is concluded that a single universal maximum limit on the atrazine application in catchments, as suggested by many regulatory authorities, does not provide adequate protection of the aquatic environment. Rather, it is advocated that flexible limits on the application of atrazine be developed in line with the potential risk of contamination to surface and subsurface water and fragility of the aquatic environment. PMID- 11485217 TI - Effects of brine injection wells, dry holes, and plugged oil/gas wells on chloride, bromide, and barium concentrations in the Gulf Coast Aquifer, southeast Texas, USA. AB - Data from 1,122 brine injection wells, 24,515 dry holes, 20,877 plugged oil/gas wells, and 256 water wells were mapped with a geographic information system (GIS) and statistically analyzed. There were 9, 107, and 58 water wells within 750 m of a brine injection well, dry hole, or plugged oil/gas well, respectively. Computed median concentrations were 157 mg/l for chloride, 0.8 mg/l for bromide, and 169 microg/l for barium. The maximum chloride concentration was 2,384 mg/l, close to 10 times the secondary drinking water standard. Shallow water wells and water wells near plugged oil/gas wells had significantly higher chloride and bromide levels. PMID- 11485218 TI - Heavy metals in the aquatic environment of the Southern Adriatic Sea, Italy: macroalgae, sediments and benthic species. AB - Samples of sea urchins (Paracentrotus lividus), holothurians (Holothuria polii), green algae (Ulva lactuca. Codium vermilara and Enteromorpha prolifera) and sediments were collected from different coastal zones of the South Adriatic Sea (Italy). The occurrence of metals in macroalgae is poor especially if compared with that reported in other coastal areas affected by human activities, with the exception of Fe that showed high mean values (405 microg g(-1) dry wt.). Likewise, relationships between metal concentrations in holothurians and sediments were found, demonstrating that H. polii could serve as bioindicator for Hg and Cu. PMID- 11485219 TI - Chemical and biological evaluation of a reaction mixture of R-(+)-limonene/ozone: formation of strong airway irritants. AB - The airway irritation of a reaction mixture of R-(+)-limonene and ozone was evaluated by a mouse bioassay in which sensory irritation, bronchoconstriction and pulmonary irritation were measured. Significant sensory irritation (33% reduction of mean respiratory rate) was observed by dynamic exposure of the mice, during 30 min, to a ca. 16 s old reaction mixture of ozone and limonene. The initial concentrations were nominally 4 ppm O3 and 48 ppm limonene. After reaction, the residual O3 was <0.03 ppm. Conventional analytical chemical methods were used to measure the formation of readily identified and stable products. Besides the expected products, 1-methyl-4-acetylcyclohexene (AMCH), 3-isopropenyl 6-oxoheptanal (IPOH), formaldehyde and formic acid, autooxidation products of limonene and a series of compounds including acetone, acrolein and acetic acid, which may or may not be artefacts, were identified. Addition of the sensory irritation effects of the residual reactants and all the identified compounds could not explain the observed sensory irritation effect. This suggests that one or more strong airway irritants were formed. Since limonene is common in the indoor air, and ozone is infiltrated from outdoors and/or produced indoors (e.g., by photocopiers), such oxidation reactions may be relevant for indoor air quality. PMID- 11485220 TI - Assessing relocation strategies of urban air quality monitoring stations by GA based compromise programming. AB - This paper presents a GA-based compromise programming technique for assessing the relocation strategy of urban air quality monitoring network with respect to the multi-objective and multi-pollutant design criteria. While the impact of conservative, quasi-stable, and reactive pollutants are considered in the design principles via a simulation analysis, cost, effectiveness, and efficiency characteristics are postulated in the optimization process. Therefore, technical coverage for illustrating the needs of siting air quality monitoring stations (AQMS) includes both the air quality simulation and optimization modeling analyses in a two-stage analytical framework simultaneously. It starts from determining the spatial interrelationship among those candidate sites using various types of air quality simulation models as an integrated means. And the outputs drawn from the simulation models can then be used as the required inputs in the compromise programming model in order to screen all those siting alternatives that may satisfy the planning goals subject to the essential constraints throughout the multi-objective optimization process. For the illustrating purposes, a series of technical settings for finding the optimal relocation scenarios of AQMS were examined in the case study for the city of Kaohsiung in South Taiwan where the long-term violations of official standards of ozone and particulates turn out to be critical. It not only expresses the ideas of relocation strategy but also indicates how to utilize those alternatives in the decision-making process for improving the functionality of air quality monitoring in the urban environment. Experience gained in this study clearly indicates that the more the number of pollutants and objectives considered simultaneously, the higher the number of candidate sites to be selected in the relocation strategy. PMID- 11485221 TI - Trace elements determination in lichens and in the airborne particular matter for the evaluation of the atmospheric pollution in a region of northern Italy. AB - Lichens as biomonitors and neutron activation analysis as analytical technique have been employed to study the distribution of trace elements (TE) in a mountain region of north Italy (Biella) characterized by settlements of wool industry. Samples of airborne particulate matter collected onto filters, different species of lichens and samples of soils have been analyzed for the calculation of the enrichment factors (EFs) of more than 25 TE. By comparison of the corresponding EFs, the most suitable lichen species (Parmelia caperata) was selected as specific TE biomonitor of the area investigated. Samples of this lichen were collected and analyzed for the evaluation of the TE distribution in four different locations. The results obtained from the analysis of the lichens gave information about the predominant direction of pollutants transportation, while those related to the concentrations found in the air particulate allowed the evaluation of the degree of the local TE atmospheric pollution. PMID- 11485222 TI - Integrated measures for preservation, restoration and improvement of the environmental conditions of the Lagoon Olho d'Agua basin. AB - The Lagoon Olho d'Agua in Pernambuco State, Northeast Brazil has received increasing environmental concern due to significant stress from pollution in the catchment. The existing environmental problems are the result of great pressure from a broad range of human activities, especially in the last 10 years. Serious pollution exists mainly from some industrial and urban activities, which increased intensively after the eighties. There is a strong social and economical pressure for housing and construction near the lagoon, due to the available land nearby beaches and estuarine zone, and recently by growing tourism activities. Uncontrolled land use by low-income communities and the pressure for construction by developers have led to landfilling and to deterioration of water quality in the lagoon catchment. Improvement of the environmental conditions in the catchment needs integrated measures. Guidelines and some specific actions involving several institutions have been established and refer to sanitation and urban infrastructure as the main priorities. A main target is the construction of low-cost sewage system with smaller and decentralised treatment plants. PMID- 11485223 TI - Concentrations of natural radionuclides in imported mineral substances. AB - Levels of natural radionuclides, such as thorium-232 (232Th), radium-226 (226Ra) and potassium-40 (40K), were determined in about 2,000 samples of imported mineral substances using gamma spectrometry. The maximum concentration levels of 232Th, 226Ra and 40K were 1,260 +/- 54, 13,400 +/- 94 and 1,256 +/- 260 Bq kg( 1), respectively, in zirconium compounds. Due to relatively high concentrations of the above-mentioned radionuclides in some imported mineral substances, the National Radiation Protection Department (NRPD), as the competent authority, has established national limits (900 Bq kg(-1) for 232Th, 1800 Bq kg(-1) for 226Ra and 11,000 Bq kg(-1) for 40K), based on the external exposures, for general importation permits of these materials. PMID- 11485224 TI - The study of Central Taiwan particles concentration variations during earthquake period. AB - Ambient particle concentration was taken on the traffic sampling site over the Chung-Chi Road over the bridge (CCROB) in front of Hungkuang Institute of Technology (HKIT). The sampling time was from August 1999 to December 1999. During the sampling period, Taiwan's biggest earthquake in more than a century registered 7.3 on the Richter scale (Taiwan Chi-Chi Earthquake). Besides, there are more than 20,000 aftershocks following the Taiwan Chi-Chi Earthquake within 3 months. Thus, the mass concentration of particles with aerodynamic diameters smaller than 2.5 microm (PM2.5) and PM2.5-10 was also collected then compared with the total mass concentration of suspended particles (TSP) in this study. The average TSP, PM2.5-10, and PM2.5 concentrations are 106, 24.6, and 58.0 microg/m3, respectively, after the Taiwan Chi-Chi Earthquake. The average TSP concentrations before and after Taiwan Chi-Chi Earthquake were 69.6 and 127 microg/ m3, respectively. In addition, statistical analysis of the PM10 data from this study and EPA in 1999 yielded a Tstatistic of 0.147, which is smaller than t(0.975,18) = 2.101. It is indicated that there was no significant difference. So, the PM10 concentrations measured after Taiwan Chi-Chi Earthquake in this study were also greater than those data previously obtained from Taiwan EPA in the same region of this area. The relationships between TSP, PM10, PM2.5-10, and PM2.5 particle concentrations and wind speed (R2) are .77, .59, .58, .58, respectively. And the ratios of PM2.5/PM25-10, PM2.5/PM10, and PM10/TSP are 221%, 67.2%, 58.0%, respectively. The average ratios of PM2.5/PM2.5-10 and PM2.5/PM10 increase by about 120% and 17%. It indicated that the fine-particles concentration increases compared to the coarse-particles concentration after 921 Taiwan Chi-Chi Earthquake. And the proposed reasons are that local motor vehicle emissions combined the fine particles transported from the Chi-Chi epicenter. More importantly, the wind direction was mainly blown from southeastern part. These two main factors enhance the fine-particles concentration in this area. PMID- 11485225 TI - Assessment of groundwater quality impacts due to use of coal combustion byproducts to control subsidence from underground mines. AB - Coal combustion byproducts are to be placed in an underground coal mine to control subsidence. The materials were characterized to determine potential groundwater impacts. No problems were found with respect to heavy or toxic metals. Coal combustion byproduct leachates are high in dissolved solids and sulfates. Chloride and boron from fly ash may also leach in initially high concentrations. Because the demonstration site is located beneath deep tight brine-bearing aquifers, no problems are anticipated at the demonstration site. PMID- 11485226 TI - Setting up the criteria and credit-awarding scheme for building interior material selection to achieve better indoor air quality. AB - Methods, standards, and regulations that are aimed to reduce indoor air pollution from building materials are critically reviewed. These are classified as content control and emission control. Methods and standards can be found in both of these two classes. In the regulation domain, only content control is enforced in some countries and some regions, and asbestos is the only building material that is banned for building use. The controlled pollutants include heavy metals, radon, formaldehyde, and volatile organic compounds (VOCs). Emission rate control based upon environment chamber testing is very much in the nature of voluntary product labeling and ranking, and this mainly targets formaldehyde and VOC emissions. It is suggested that radon emission from building materials should be subject to similar emission rate control. A comprehensive set criteria and credit-awarding scheme that encourages the use of low-emission building material is synthesized, and how this scheme can be practiced in building design is proposed and discussed. PMID- 11485227 TI - Antimicrobial resistance among Pseudomonas spp. and the Bacillus cereus group isolated from Danish agricultural soil. AB - From four Danish pig farms, bacteria of Pseudomonas spp. and the Bacillus cereus group were isolated from soil and susceptibility towards selected antimicrobials was tested. From each farm, soil samples representing soil just before and after spread of animal waste and undisturbed agricultural soil, when possible, were collected. Soil from a well-characterized Danish farm soil (Hojbakkegaard) was collected for comparison. The Pseudomonas spp. and B. cereus were chosen as representative for Gram-negative and Gram-positive indigenous soil bacteria to test the effect of spread of animal waste on selection of resistance among soil bacteria. No variations in resistance levels were observed between farms; but when the four differently treated soils were compared, resistance was seen for carbadox, chloramphenicol, nalidixan (nalidixic acid), nitrofurantoin, streptomycin and tetracycline for Pseudomonas spp., and for bacitracin, erythromycin, penicillin and streptomycin for the B. cereus group. Variations in resistance levels were observed when soil before and after spread of animal waste was compared, indicating an effect from spread of animal waste. PMID- 11485228 TI - Determination of 2-butoxyethanol emissions from selected consumer products and its application in assessment of inhalation exposure associated with cleaning tasks. AB - Consumer products are important sources of human exposure to certain chemicals. Recent regulatory requirements for assessing human exposure to three glycol ethers, namely 2-methoxyethanol (ME), 2-ethoxyethanol (EE) and 2-butoxyethanol (BE), have prompted the investigation of these chemicals in consumer products and their emission characteristics. Thirteen products were selected for investigation based on their potential of containing the chemicals. Headspace results indicated that ME and EE were not present in any of the 13 selected products, while BE was detected in the headspace samples of seven products, of which five were household cleaning agents. Other related compounds such as 2-hexyloxyethanol (HE) and 2-(2 butoxyethoxy)ethanol (BEE) were also detected in the headspace samples of some products. BE emissions from five cleaning related products were measured using a field and laboratory emission cell (FLEC) with its subunit to provide emission data for inhalation exposure assessment purposes. These products had initial emission factors ranging from 145 to 938 mg m(-2) h(-1) under the experimental conditions. It was found that the emission factor of BE was inversely proportional to the dilution factor of the products. A good relationship was established between the emission factor of BE and its concentrations in water based products. Based on product use scenarios developed by US EPA and an assumed "standard room," average daily inhalation exposure levels of a resident as a result of performing cleaning tasks were estimated to be 0.075 and 0.186 mg (kg b.w.)(-1) day(-1) for two all-purpose spray cleaners, and 0.004 and 0.006 mg (kg b.w.)(-1) day(-1) for two-spray glass cleaners, respectively. PMID- 11485229 TI - Bovine pericardium for duraplasty: clinical results in 32 patients. AB - Bovine pericardium has been widely used for grafts in cardiac surgery and seems to have suitable properties for use as a dural graft. We report on the use of solvent-preserved, gamma-sterilized Tutoplast bovine pericardium for dural grafts in 32 patients undergoing cranial and spinal operations with the objective of clinically assessing this material and technique by a retrospective analysis. All available records were reviewed and information regarding the indication for grafting, complications, and outcome were collected and analyzed for all patients. Indications for grafting included tethered cord myelolysis, closure of lumbosacral myeloceles, Chiari decompression, posterior fossa craniotomy, supratentorial craniotomy, and trauma. Outcomes were excellent in 31 patients; the one poor outcome was unrelated to surgical closure. The dural graft was not intended for outcome in any patient. Bovine pericardium was found to be a flexible and easily suturable, safe and cost-effective material for duraplasty. These results confirm the excellent suitability of Tutoplast bovine pericardium for dural substitution. PMID- 11485230 TI - Surgical treatment of thalamic hematomas via the contralateral transcallosal approach. AB - Acute management of deep-seated hematomas remains controversial. Since patients with these hematoma later tend to develop severe edema and necrosis around the lesion, when surgery is indicated it should be done as early as possible. The purpose of this study was to compare whether early surgical removal and conservative treatment of primary thalamic hematoma correlated with improved neurological outcome. Last year, 61 patients with primary thalamic hematomas were admitted to our institution. Of these, 21 underwent surgery via contralateral transcallosal approach during the ultraearly stage (within 6 hours) after the apoplectic attack, and 24 patients were treated conservatively. Another 16 patients were excluded from the study due to systemic disease, mild hematoma (<40 cc), and deep coma associated with absence of brain stem reflexes. Initial Glasgow coma scores (GCS) at admission were similar for operated and nonoperated patients (8.64 +/- 1.93 versus 9.50 +/- 2.10, P>0.05). In the operated group, two patients had good recoveries and returned to normal life (Glasgow Outcome Score, or GOS, I), four had moderate disability and needed partial care (GOS II), six had severe disability and needed nursing care (GOS III), and six had a vegetative state (GOS IV). However, in the nonoperated group, one patient had good recovery and returned to normal life (GOS I), two had moderate disability and needed partial home care (GOS II), three had severe disability and needed nursing care (GOS III), and six had a vegetative state (GOS IV). In this group, the 30-day mortality rate was 50%. Mortality was markedly lower in the operated group (14.3%) than the nonoperated group. and this difference was statistically significant (chi2=3.33, P<0.05). From this study, we believe that evacuation of primary thalamic hematoma via the contralateral transcallosal microsurgical approach may be useful for deciding on the indication and predicting the functional prognosis. PMID- 11485231 TI - Intracerebral ganglioglioma: clinical and radiological study of eleven surgically treated cases with follow-up. AB - BACKGROUND: Gangliogliomas are rare benign tumors of the CNS consisting of differentiated neural elements and low-grade glial cells. METHODS: We reviewed our experience of 11 patients with histologically proven ganglioglioma who were surgically treated since 1986 at Cukurova University Medical Center. These patients presented at 18 to 45 years of age. Five were women and six were men. The most common initial symptom was seizures (in nine of 11 patients), which had sometimes persisted over long periods of time. At the time of diagnosis, four patients had focal neurological deficits and three had signs or symptoms of increased intracranial pressure. The cystic and well-circumscribed characteristics of these lesions were detected on computed tomography (CT). Despite their appearance on CT, all but one of the lesions were found to be mostly solid at operation. Magnetic resonance imaging (MRI) in six patients revealed abnormally high signal intensity on T2-imaging. The temporal lobe was the main tumor location (seven patients). All cases were diagnosed according to the Russel and Rubinstein histological criteria for ganglioglioma. RESULTS: Ten patients had radical total resection and one had subtotal resection. No patient underwent postoperative radiation or chemotherapy. Except for one, all are still alive and free of progressive disease 1 to 11 years (mean 6.2) after operation. Six are seizure-free and three have improved seizure control under anticonvulsant therapy. CONCLUSIONS: We conclude that ganglioglioma is a distinct histological phenomenon with mildly predictable clinical symptoms (seizures), mildly characteristic radiological features, and long-term survival after surgical resection without the need of adjuvant treatment such as radiotherapy. PMID- 11485232 TI - Use of an autologous cortical bone graft sandwiched between two intervertebral spacers in posterior lumbar interbody fusion. AB - Various intervertebral spacers with or without posterior instrumentation use pedicle screw fixation in posterior lumbar interbody fusion (PLIF). Recently we harvested an autologous cortical bone graft from a spinous process by en bloc resection and inserted it between two intervertebral spacers during PLIF surgery. Due to better balance, this procedure provides greater mechanical strength, larger contact area, and better bilateral restoration of disc height than PLIF using intervertebral spacers only, and there is no need to take a bone graft from the iliac crest. This technique appears to result in effective spinal fusion in PLIF surgery. PMID- 11485233 TI - Intraoperative ultrasonographically guided direct ethanol injection for a brain metastasis from renal cell carcinoma: case report. AB - A 60-year-old man presented with a brain metastasis from renal cell carcinoma (RCC). Computer tomography revealed a homogeneously enhanced tumor, 3.0 cm in maximum diameter, in the right medial temporal lobe. Cerebral angiography revealed that the tumor was mainly fed by the right posterior cerebral artery. Surgery was performed via right temporal craniotomy. After craniotomy, under ultrasonography (US) guidance, a total of 3.0 ml of ethanol was injected into the tumor to diminish the intratumoral vascular flow. Ultrasonographic guidance was very useful in monitoring the intratumoral vascular flow. After a marked decrease in the vascular flow, the tumor was totally removed using piecemeal technique. During surgery, only minimal bleeding from the tumor was noted. The postoperative course was uneventful, with no evidence of injected ethanol-related complications. In conclusion, this can be a safe, easy, and effective therapeutic technique for diminishing vascular flow within brain tumors rich in vascularity, such as brain metastases from RCC. PMID- 11485234 TI - Nimodipine attenuates lipid peroxidation during the acute phase of head trauma in rats. AB - Oxygen free radical-mediated lipid peroxidation is one of the major mechanisms of secondary damage in traumatic brain injury. We assessed the effects of nimodipine on lipid peroxidation 1 h after head trauma in rats. Nimodipine (1.5 microg/kg IV bolus injection) was given immediately after head trauma by either the carotid artery or the jugular vein. Placebo treated rats received saline by the same routes. Control rats received head trauma only. Sham-operated rats were the group without head trauma. Malondialdehyde (MDA), which is the end product of lipid peroxidation, was measured as an indicator of oxygen free radical formation in the brain tissue. The mean values for MDA in sham operated rats were 92.4 +/- 4.9 nanomoles/gram wet weight (nmol/gww) of brain tissue. In the control group, MDA content of the brain tissue was 120.8 +/- 9.4 nmol/gww. In placebo treated rats, the results were similar. In the groups receiving nimodipine via carotid artery or jugular vein, the mean values were 101.1 +/- 6.9 and 106.5 +/- 6.0 nmol/gww, respectively. These results indicate that nimodipine caused a significant decrease in lipid peroxidation when given in the acute phase of head trauma in rats. This occurred regardless of the route of injection. PMID- 11485235 TI - The effect of allopurinol on focal cerebral ischaemia: an experimental study in rabbits. AB - In this experimental study, the neuroprotective effect of the xanthine oxidase inhibitor allopurinol on focal cerebral ischaemia created by permanent middle cerebral artery occlusion (MCAO) was investigated. Using high performance liquid chromatography (HPLC), we measured hypoxanthine, xanthine, and uric acid (UA) levels in rabbit brains following focal cerebral ischaemia. Rabbits were randomly and blindly assigned into four groups of eight animals each. The control groups received 2% carboxymethylcellulose solution, while 10% allopurinol 150 mg/kg was given to the treatment group 1 h before ischaemia. Each group was subdivided into two groups which were sacrificed 4 h or 24 h after ischaemia, respectively. UA and xanthine values of the rabbits in the control groups were quite high at both times and highest after 24 h, particularly in the centre of the ischaemia. A significant decrease in UA and xanthine values was observed in rabbits that were given allopurinol (P<0.05). According to our results, it was concluded that allopurinol pretreatment protects neural tissue in the early period after arterial occlusion and prevents cerebral injury in the late period, especially in the perifocal area, possibly by preventing the formation of free radicals with xanthine oxidase inhibition. PMID- 11485236 TI - Effects of trapidil on ATPase, lipid peroxidation, and correlation with ultrastructure in experimental spinal cord injury. AB - The present study was performed to investigate the effect of trapidil on ischemic damage of cells after spinal cord injury. The injury was produced by extradural clip compression of the exposed spinal cord in rats according to Rivlin and Tator. The ten rats in group 1 were used to determine normal findings without any surgery or medication. On the 15 rats in group 2, only six-level laminectomy was performed to determine the influence of the total laminectomy on the biochemical factors measured and the, light and ultrastructural findings. The 15 rats each in groups 3 and 4 were used as trauma and trapidil (40 mg/kg) treatment groups, respectively. The injury actually produced a significant decrease in Na+-K+/Mg+2 ATPase activity of the injured segments as early as 10 min after trauma. Trapidil attenuated Na+-K+/Mg+2 ATPase inactivation in the traumatized rats for 120 min after treatment (P<0.05) and significantly reduced the malone dialdehyde content below that in the traumatized group at all determined times (P<0.05). Light and electron microscopic findings supported the biochemical results. PMID- 11485237 TI - Posterolaterally displaced type IIA odontoid fractures. AB - A case of type IIA odontoid fracture with posterolateral dislocation accompanied by spinal cord injury is presented. Cervical traction was employed but reduction could not be achieved with up to 8 kg of traction. The patient was treated with intraoperative reduction and C1-2 posterior transarticular screw fixation with supplemental bone-wire fusion, and rigid fixation was obtained without any complication. PMID- 11485238 TI - A modified burr-hole method in galeoduroencephalosynangiosis for an adult patient with probable moyamoya disease--case report and review of the literature. AB - A 49-year-old woman with probable moyamoya disease was surgically treated by combined direct and indirect methods, superficial temporal to middle cerebral artery anastomosis and galeoduroencephalosynangiosis by a burr-hole method developed by Kawamoto et al. Transient ischemic attacks and motor weakness of bilateral lower extremities disappeared completely within 1 month and never recurred during 1-year follow-up. Galeoduroencephalosynangiosis by a burr-hole method appears useful for preventing ischemic damage of the territory of the anterior cerebral artery in adult patients with probable moyamoya disease. PMID- 11485239 TI - Giant pericallosal artery aneurysm: case report and review of the literature. AB - Pericallosal artery aneurysms comprise 5% of all intracranial saccular aneurysms and are usually small. Giant cerebral aneurysms mostly occur in major arteries. To date, 12 cases of giant pericallosal artery aneurysm have been reported in the literature. An unusual giant thrombosed pericallosal artery aneurysm is reported here. A 65-year-old female presented with headache and personality changes. Computed tomography, magnetic resonance imaging, and cerebral angiography revealed a right-sided giant thrombosed pericallosal artery aneurysm. The patient was operated via an anterior interhemispheric approach and the neck of the aneurysm was successfully clipped. The postoperative period was uneventful. This rare lesion is one of few cases presented in the literature in which neuroradiologic and neuropathologic evaluation was completely performed and the neck of the aneurysm was clipped. PMID- 11485240 TI - Spinal epidural abscess: a meta-analysis of 915 patients. PMID- 11485241 TI - Central nervous system cavernomas in the pediatric age group. AB - Pediatric CNS cavernomas still are diagnostically and therapeutically challenging lesions. With the help of magnetic resonance imaging, the natural history of cavernomas now guiding therapeutic strategies is well documented in adults but remains poorly known in the pediatric age group, since most previous studies dealt with adult and pediatric patients together. This paper focuses on clinical, imaging, and therapeutic features and differential diagnosis of CNS cavernomas with an emphasis on their specificities in the pediatric age group. It is based upon a critical review of the literature and our single-center experience with 36 children (35 with cerebral cavernomas and one with spinal cord cavernoma) operated on during the period of 1985-1999 as well as with seven additional unoperated pediatric cases. Our experience resembles that of other authors regarding the high hemorrhagic risk in children compared to adults. These angiographically occult vascular malformations are often revealed by the sudden onset of intracerebral hematoma with acute focal neurologic deficits, concomitant manifestations, and/or signs of raised intracranial pressure. True epilepsy is less common and may be related to chronic or recurrent microbleeding. Evocative imaging findings are also somewhat different in the two age groups, and we propose here an imaging classification of cerebral cavernomas based on both morphological and signal characteristics that is applicable to the pediatric age group. A sharply demarcated spherical intracerebral hematoma or heterogeneous lesion should always make one consider the hypothesis of a cavernoma. For symptomatic lesions and most rapidly growing asymptomatic lesions, the treatment of choice is complete microsurgical excision preceded by careful anatomical and functional evaluation. Improvements in surgical techniques and anesthesiology over recent years have brought good results in most operated children. The limited role of radiosurgery in the management of pediatric cerebral cavernomas is discussed. There is still a need for well-conducted specific evaluation of the natural history of these lesions in the pediatric age group to aid in systematic research, follow-up, and therapeutic strategies for asymptomatic cavernomas. PMID- 11485242 TI - In vivo experiences with frameless stereotactically guided screw placement in the spine--results from 75 consecutive cases. AB - Whereas cranial neuronavigation is widely accepted as a helpful tool, larger series of the in vivo application of spinal neuronavigation do not exist. In the following we report our 4-year experience with spinal navigation in 75 consecutive cases for dorsal transpedicular screw placement. Seventy-five patients were planned for operation employing anatomical reference points defined on a 2-mm high resolution CT. We used single vertebra registration and surface matching. With the above methods, the mean registration deviation ranged from 0.18 mm (cervical spine) to 0.31 mm (lumbar spine). All our screws in the upper cervical spine were navigated correctly (17 patients), thus improving markedly the surgical outcome. The results were not as promising in the lumbar area. In only 84% was navigation reliable. The reason was the lack of a practicable tracking tool. Spinal neuronavigation based on anatomical reference points is able to improve the results in transpedicular screwing, especially in the cervical spine. The lack of a practicable tracking tool still hinders its use in routine clinical application. PMID- 11485243 TI - Surgical therapy for lumbar spinal stenosis: evaluation of 300 cases. AB - Although lumbar spinal stenosis syndrome is extremely common, considerable controversy continues to surround its classification, diagnosis, and treatment. In this article, we retrospectively analyzed the clinical, radiological, and surgical characteristics of 300 patients operated on in our clinics with the diagnosis of lumbar spinal stenosis. Surgical therapy and outcome are discussed with respect to the known facts. After clinical and radiological evaluation, 300 patients were diagnosed with lumbar spinal stenosis. All patients underwent laminectomy and bilateral foraminotomy at the stenotic level. Surgical outcomes were evaluated as very good, good, same, and poor according to the clinical findings. Ninety percent of the surgically treated patients demonstrated very good and good outcomes. Our conclusion in lumbar spinal stenosis diagnosed by objective clinical and radiological findings is that decompressive laminectomy and extensive foraminotomy without instrumentation allow a good outcome. PMID- 11485244 TI - A new method for surgical repair of impression fractures of the cranial vault and frontal sinus with rivet-like titanium clamps. AB - Conventional methods of stabilizing fragments in cranial impression fracture include the use of threads, wires, plates, etc. A new approach to facilitate this surgery is the use of titanium clamps, presented here in five cases, including one with frontal sinus fracture. Surgery was performed on admission day, with the exception of the sinus fracture. Compared with the use of mini- and microplates, the procedure was simple and short. The fixation was very rigid and the esthetic result excellent in all five cases. Skin irritation was minimal, compared with that from miniplates. Computed tomography and MRI compatibility due to artifacts are identical to those with miniplates. PMID- 11485245 TI - Spinal intramedullary metastases. Report of two cases and review of the literature. AB - Two cases of intramedullary metastasis were observed in our department within a short period of time that change the picture of uncommon locations of spinal metastases as described in literature. The patients presented primarily with rapidly progressing flaccid paraparesis of the lower extremities. Both additionally described diffusely located hypesthesia and pallhypesthesia of the limbs. One complained about sphincter disturbance. Both were treated by laminectomy and microsurgical extirpation of the tumor. In the first case reported here, an intramedullary metastasis of an epithelioid sarcoma in the conus medullaris region was ascertained which may be regarded as the first such reported case. In the second case report, histologic findings confirmed a thoracic intramedullary metastasis of a mastocarcinoma. At follow-up 6 and 8 months postoperatively, we observed full recovery of neurologic function in one patient and partial recovery in the other. Intramedullary metastases are extremely rare. Clinical presentation may show rapid onset and outcome is related to rapid resection and adjuvant therapy. In contrast to intracerebral metastases, this is a remarkable incidence of sarcomatous intramedullary metastatic disease. PMID- 11485246 TI - Trigeminal root recording in normal trigeminal function. AB - Despite many investigations on the detection of trigeminal evoked potentials (TEP), there is still no consensus on a technique or interpretation of its components. In order to obtain clear TEP waveforms and analyze the origin of early components, we recorded TEP by the conventional far-field technique at the ipsilateral auricula as well as by near-field technique with a bipolar electrode at the trigeminal root just distal to the entry zone of its root. Trigeminal recordings were performed in nine patients with intrameatal vestibular schwannomas and in three with trigeminal neuralgia without trigeminal nerve deficits. In near-field recording at the trigeminal root, a triphasic response was recorded, with the first component occurring at 3.16 ms, 2.98 ms, and 4.16 ms following supraorbital, infraorbital, and mandibular nerve stimulation, respectively. Using conventional farfield TEP, the first component recorded at the trigeminal root occurred later than the first component in all other recordings. This suggests that the N2.0 component of far-field TEP is of extra axial origin and the N5 component is of brainstem origin. The N13.0 component may be of cortical origin. We conclude that the N5.0 component and N5.0-N13.0 interpeak latency may be used as electrophysiological parameters of brainstem function during posterior fossa surgery. PMID- 11485247 TI - Is exon 5 of the PTEN/MMAC1 gene a prognostic marker in anaplastic glioma? AB - Chromosome 10 deletions are among the most common genetic changes in highly malignant glial tumors. It has been noted that loss of heterozygosity (LOH) at 10q23 is a frequent alteration in a variety of human tumors and occurs in approximately 70% of all glioblastomas. By mapping of homozygous deletions on 10q23, a candidate tumor suppressor gene has been isolated, called PTEN for "phosphatase and tensin homolog deleted on chromosome 10" and MMAC1 for "mutated in multiple advanced cancers-1." Mutations of this tumor suppressor gene PTEN/MMAC1 have been reported in anaplastic glial tumors. The objective of this paper was to individuate a prognostic marker in exons 5, 6, 7, and 8 of the PTEN/MMAC1 gene for the high-grade malignant glioma with the most aggressive clinical behavior. In this study, we undertook sequence analysis of these exons in six selected patients with high-grade malignant gliomas who underwent radical aggressive tumor resection followed by radiotherapy within 3 weeks after surgery and subsequent chemotherapy. In them, the exon 5 sequence of the PTEN/MMAC1 gene is suggestive of a genetic survival marker in gliomas with high-grade malignancy. PMID- 11485249 TI - Detoxification of endotoxin by endodontic irrigants and calcium hydroxide. AB - The effects of endodontic irrigants and calcium hydroxide on lipopolysaccharide (LPS; endotoxin) were analyzed using the highly selective technique of mass spectrometry/gas chromatography with selected ion monitoring. An aqueous solution of LPS was mixed with one of a variety of endodontic irrigants for 30 min. Because it is a commonly used interappointment dressing, calcium hydroxide was also applied to LPS for 1, 2, or 5 days. LPS inactivation was measured by quantitation of free fatty acid release. Water, EDTA, ethanol, 0.12% chlorhexidine, chlorhexidine + sodium hypochlorite, and sodium hypochlorite alone showed little breakdown of LPS. Long-term calcium hydroxide--as well as 30-min exposure to an alkaline mixture of chlorhexidine, ethanol, and sodium hypochlorite--did detoxify LPS molecules by hydrolysis of ester bonds in the fatty acid chains of the lipid A moiety. PMID- 11485248 TI - Evaluation of regional bond strength of resin cement to endodontic surfaces. AB - The purpose of this study was to test the feasibility of adapting a new microtensile testing technique to measure resin cement bond strengths to the cervical, middle, and apical thirds of root canals. Post spaces were created in extracted human teeth, and the roots were ground flat on one side to expose the canal and permit ideal placement of one of two resin cements (Panavia 21 or C&B Metabond). After 48 h of storage, serial 1-mm-thick cross-sections were cut to create 6-10 specimens per root. The first three specimens were from the cervical third, the next three were from the middle third, and the last three were from the apical third of the root. Each 1 x 1 x 8 mm specimen was pulled to failure in a miniature testing machine. The results indicated that both resin cements produced high bond strengths (12-23 MPa), and that bond strengths to the apical third were significantly higher (p < 0.05) than to the cervical or middle third with either cement. This new method shows promise for evaluating resin bond strengths within root canals. PMID- 11485250 TI - Antimicrobial activity of Ca(OH)2 containing pastes with Enterococcus faecalis in vitro. AB - Sixty-eight standardized human root specimens were infected with Enterococcus faecalis for 3 wk after removal of the smear layer. After 3 wk of infection, the smear layer was reformed and in half of the specimens, the smear layer was again removed. Aqueous Ca(OH)2 paste and silicone oil based Ca(OH)2 paste were used as the test medications. The specimens were divided into four groups (i.e. (a) nonsmeared aqueous calcium hydroxide group, (b) nonsmeared silicon oil-based calcium hydroxide group, (c) smeared aqueous calcium hydroxide group, and (d) smeared silicon oil-based calcium hydroxide group. Medications were placed in the canals for 7 days. After removal of medications dentin chips were collected and incubated. The quantity of bacteria present was assessed. All calcium hydroxide pastes were effective in the elimination of bacteria in the dentinal tubules, except in the smeared group with silicone oil-based calcium hydroxide. PMID- 11485251 TI - Impact of torsional and bending inertia on root canal instruments. AB - The laws of applied mechanics can be used to analyze and predict the behavior of mechanical parts subjected to strain and distortion, such as endodontic instruments. Simple mathematical formulas can be used to represent the impact of torsional and bending inertias on stresses in endodontic instruments being used in root canals. The boundary integral method was applied to two theoretical models with the same diameters but different cross-sections to evaluate quantitatively and qualitatively the stresses produced in orthogonal cross sections. The results confirmed the mathematical deduction concerning the impact of torsional and bending stresses. The results indicate that clinical protocols based on the torsional and bending properties of endodontic instruments must be strictly followed, and endodontic instrument manufacturers must adopt standardized criteria for describing these properties. PMID- 11485252 TI - Ultrastructural analysis of MHC class II molecule-expressing cells in experimentally induced periapical lesions in the rat. AB - Periapical lesions were induced by making 28 days of unsealed pulp exposures in the lower first molars of Wistar rats. Major histocompatibility complex class II molecule-expressing cells were then demonstrated by means of immunoperoxidase staining using a monoclonal antibody OX6, and the ultrastructure of these cells was analyzed under electron microscopy. OX6+ cells were classified into two major populations, (i.e. macrophages and dendritic cell (DC)-like cells. DC-like cells had elongated cytoplasmic processes, contained a few lysosomal structures, lacked distinct phagosomes, and were the most predominant cell type in the established lesion. Some of lymphocytes and plasma cells also showed a positive immunoreactivity. Both OX6+ macrophages and DC-like cells often showed a cell-to cell attachment with lymphocytes. These findings suggested that major histocompatibility complex class 11 molecule-expressing macrophages and DC-like cells may play a crucial role in periapical lesion development by acting as antigen-presenting cells to memory T lymphocytes. PMID- 11485253 TI - Calcium concentration and pH of the periapical environment after applying calcium hydroxide into root canals in vitro. AB - The purpose of the present study was to determine both calcium concentration and pH in the periapical region after application of 1 of 4 different calcium hydroxide preparations into experimental root canals. Fifty root canal models were divided into five groups: group 1--calcium hydroxide was mixed with distilled water at a powder/water weight ratio of 38%; group 2--calcium hydroxide was mixed with distilled water at 44%; group 3--calcium hydroxide was mixed with distilled water at 50%; group 4--calcium hydroxide powder only was used; and group 5-the control group, in which nothing was applied to the canals. All samples were immersed in distilled water maintained at 37 degrees C. Calcium concentration and pH of the distilled water were measured after 3 days, 7 days, and then at weekly intervals up to 15 wk, during which time the storage medium was renewed after each measurement. Calcium concentration and the change in pH of the distilled water were statistically quicker and greater in groups 1 to 3 (mixture groups) than group 4 (powder only) (p < 0.05). The highest calcium concentration (peak Ca2+ release) was observed after 3 days for the mixture groups, whereas that for the powder only group was found at 7 days. Peak pH change was found after 14 days for the mixture groups, whereas that for the powder only group was found at 49 days. After peaking, all groups showed a decline of the pH over time. These results suggest that the time required for optimum intracanal activity when using calcium hydroxide mixtures is at least 2 wk. PMID- 11485254 TI - Lack of correlation among three methods for evaluation of apical leakage. AB - The purpose of this study was to compare three methods of evaluation of the apical seal. Thirty-six teeth were prepared with a ProFile device and filled according to three filling methods: Thermafil (n = 12), warm vertical condensation (n = 12), and the single cone technique (n = 2). The apical seal was recorded with three methods, each successively used on the same teeth: a fluid filtration method, an electrochemical method, and a dye penetration study. The three methods of evaluation did not rank the apical leakage of the three filling techniques in the same order. The fluid filtration method showed that the vertical condensation was superior to Thermafil, which was in turn superior to the single cone technique (p = 0.04). The dye penetration study showed that the Thermafil was a better technique than the vertical condensation and the single cone technique (p = 0.005). The electrochemical method showed no statistical difference among the three filling techniques. No correlation was found among the results obtained with the three methods of evaluation. This study shows that several studies are necessary before comparing the apical seal obtained with various filling techniques. PMID- 11485255 TI - Effect of removal of the smear layer on apical microleakage. AB - It has been shown previously that the smear layer created during instrumentation of the root canal promotes apical microleakage. The purpose of the present study was to investigate the effect of removal of the smear layer on apical microleakage, using the fluid filtration method. Thirty-six single-rooted human premolar teeth were used. Six of the teeth were selected randomly as controls, and the remaining 30 teeth were randomly divided into two groups of 15 teeth each. For the first group (group 1) teeth were irrigated with NaOCl only (smear layer not removed). In the second group (group 2) the teeth were irrigated with 15% EDTA, followed by NaOCl to remove the smear layer. Both groups were filled with thermoplasticized gutta-percha (Obtura II) using glass-ionomer cement as sealer. All teeth were stored at 37 degrees C and 100% humidity for 2 days. Apical microleakage of the filled root canal was measured by the fluid filtration technique. Hydraulic conductance was measured by the movement of an air bubble in a capillary tube after a pressure of 200 mm Hg was applied to the system. The mean microleakage rate of group 1 and group 2 were 13.0 (SD +/- 5.1) and 16.9 (SD +/- 15.3) nl/s, respectively. Removal of the smear layer caused significantly more apical microleakage (p < 0.05, Student's t test) than when the smear layer was left intact. PMID- 11485256 TI - In vitro bacterial penetration of endodontically treated teeth coronally sealed with a dentin bonding agent. AB - This study evaluated the effectiveness of a dentin bonding agent as a barrier to prevent coronal microleakage and examined the effect of a eugenol-based sealer on the sealing ability of this resin adhesive. Fifty-one extracted human mandibular molars were incorporated in a model system using an oral streptococci as a microbial marker. Group 1 consisted of 15 teeth that were obturated with only gutta-percha and received a coronal barrier of Clearfil Liner Bond 2V. Group 2 was identical to group 1, but included the use of a eugenol-based sealer in the obturation. Group 3 consisted of 15 teeth that were obturated with gutta-percha and sealer, but did not receive a coronal barrier. Six teeth served as controls. Bacterial penetration was monitored for 90 days. Results were analyzed after 30, 60, and 90 days with Fisher's exact test (p < 0.05). All controls behaved as expected. Neither group 1 nor group 2 exhibited any bacterial leakage. Eleven of the 15 specimens in group 3 leaked between 15 and 76 days. The coronal barriers in group 1 and group 2 were significantly better in preventing coronal microleakage at 60 days (p = 0.002) and 90 days (p = 0.00005). The presence of eugenol in the sealer had no significant effect on the sealing ability of Clearfil Liner Bond 2V (p = 1). PMID- 11485257 TI - Effect of spreader and accessory cone size on density of obturation using conventional or mechanical lateral condensation. AB - A simulated curved root canal in a resin block was enlarged to size 40 and used to compare the depth of accessory cone penetration and weight of obturation occurring with the use of different obturation techniques and spreader-accessory cone combinations. Twelve groups, each consisting of 10 obturations, were created. A conventional lateral condensation technique was used in six groups and a mechanical lateral condensation (MLC) technique was used in six matched groups. The six spreader-accessory cone combinations were either Fine-Medium or Fine nickel-titanium finger spreaders with either Fine, Medium-Fine, or size 25 accessory cones. Seven accessory cones were placed in every obturation. The depth of each accessory cone penetration into the canal was measured. After each obturation the gutta-percha was removed, sectioned, and the resulting mass was weighed. The means for each variable were determined and compared. MLC fills were significantly heavier and had greater depth of penetration on average than conventional lateral condensation. The best combination for heavy fills was MLC, Fine-Medium spreaders, and Fine accessory cones. The greatest mean accessory cone depth occurred with MLC, Fine-Medium spreaders, and size 25 accessory cones. PMID- 11485258 TI - Effectiveness of different obturation techniques in the filling of simulated lateral canals. AB - The purpose of this in vitro study was to evaluate the ability of lateral compaction of gutta-percha and of five thermoplasticized gutta-percha filling techniques to obturate simulated lateral canals. Sixty extracted single-rooted human teeth were instrumented at the working length to a #35 file before creating three simulated lateral canals on the mesial and distal surfaces of the root, one in each third, using a #15 engine reamer. After enlarging root canals to a #45 file, the teeth were randomly divided into six equal groups of 10 and obturated according to the following techniques: lateral compaction of gutta-percha (group A), hybrid technique (group B), Ultrafil (group C), Obtura II (group D), System B + Obtura II (group E), and Thermafil (group F). AH26 was used as the sealer. A greater number of simulated lateral canals were obturated when Ultrafil, Thermafil, and System B + Obtura II were used, in comparison with canals obturated with the hybrid technique, Obtura II, or lateral compaction of gutta percha. This difference was statistically significant (p < 0.05). No statistically significant differences were found between results obtained in the obturation of simulated lateral canals in the different thirds of the root (p > 0.05). PMID- 11485259 TI - Elimination of intracanal metallic obstructions by abrasion using an operational microscope and ultrasonics. AB - Intracanal metallic obstructions that block any further treatment often prevent the clinician from achieving total cleanliness of the root canal system. For the retrieval of these obstructions, most of the methods described suggest creating and enlarging a space around the obstruction to loosen it before its retrieval. This article describes a technique that consists of mechanical abrasion of the obstruction without altering canal walls by using an operational microscope and ultrasonic spreader. PMID- 11485261 TI - Ask a friend case #10. PMID- 11485262 TI - Effects of pressure and measurement time on the fluid filtration method in endodontics. AB - The purpose of this study was to evaluate the influence of the measurement time and the pressure on the outcome of a fluid filtration test. Thirty-six roots were prepared with a Profile device and filled with Thermafil. The leakage along the filling material was recorded under a 15 cm H2O pressure (n = 18) or a 150 cm H2O pressure (n = 18) at 3 min, 1 h, and 24 h. The fluid flow expressed as microl min(-1) decreased with an increased measurement time (p = 0.001) and was higher in the high pressure group (p = 0.001). The fluid flow of this study, where all of the teeth were filled with the same technique, ranged from 3.8 x 10(-5) microl min(-1) cm H2O(-1) to 43 x 10(-5) microl min(-1) cm H2O(-1), depending on the pressure and measurement time. This shows the need for standardization to compare the results from various studies. PMID- 11485263 TI - Cytotoxicity testing with three-dimensional cultures of transfected pulp-derived cells. AB - SV40 large T-antigen-transfected bovine pulp-derived cells were grown three dimensionally on polyamide meshes. For optimal cell growth, various cell numbers and mesh coatings were tested. Next the three-dimensional cultures were used in a dentin barrier test device, and the system was evaluated by testing a set of dental filling materials. After 24 hr exposure with or without perfusion of the pulpal part of the test device, cell survival was evaluated using an MTT assay. In all experiments pulp-derived cells transfected with SV40 large T-antigen grew three-dimensionally on polyamide meshes and showed growth kinetics similar to those on cell culture plates with lag, log, and plateau phases (reached after about 14 days of incubation). Cross-sections of the three-dimensional cell cultures revealed about 15 to 20 cell layers. In vitro cytotoxicity tests resulted in cell survival rates which are in good agreement with in vivo data and with results obtained from cytotoxicity tests with three-dimensional cultures of human foreskin fibroblasts. PMID- 11485264 TI - Glycol methacrylate: an alternative method for embedding subcutaneous implants. AB - The aim of this investigation was to study the use of glycol methacrylate (GMA) as an embedding material for rat subcutaneous tissue, which received implants of tubes with endodontic sealer. After fixation, the specimens were dehydrated in a growing alcohol series up to 95%, immersed in infiltration GMA solution and then in embedding solution. The blocks were cut into 3.0 microm sections and stained with hematoxylin-eosin. The quality of cell definition and staining allowed a quantitative analysis of the cells infiltrated in the end of the tubes. It was also possible to identify each type of inflammatory cell. Moreover, it was possible to distinguish clearly chronic from acute inflammatory cells. The GMA technique is easy to execute and reproducible, and provides a better definition of tissue cells, thus permitting definition of the degree of the inflammatory process. Therefore, it is an excellent alternative for the evaluation of the biocompatibility of endodontic sealers. PMID- 11485265 TI - Immunohistochemical localization of osteopontin in human pulp stones. AB - The organic matrix component of human pulp stones was investigated by immunohistochemistry. Two pulp stones were extracted from the upper molar teeth of two patients suffering from irreversible pulpitis. Both were formed in the center of the pulp cavity and located apart from the dentin walls. After demineralization, serial sections of the stones were prepared and subjected to immunohistochemical procedures using specific antibodies to type I collagen and noncollagenous proteins (osteopontin, osteonectin, and osteocalcin), which are reported to be involved in calcified matrix formation. Type I collagen was localized evenly in the stones, indicating that it is a major matrix component of pulp stones. Strong immunostaining of osteopontin appeared in the peripheral area of the stones, whereas osteonectin and osteocalcin were not detected. We previously reported that dental pulp cells produced osteopontin in vitro. Osteopontin has been commonly found in other pathological calcification, such as urinary stones, atherosclerotic plaques, and dental calculus. Taken together, the present findings suggest that osteopontin produced by dental pulp cells is possibly associated with calcification of the pulp stone matrix. PMID- 11485266 TI - Regulation of interleukin-6 expression in human dental pulp cell cultures stimulated with Prevotella intermedia lipopolysaccharide. AB - Interleukin (IL)-6 expression in human dental pulp cell cultures after stimulation with Prevotella intermedia lipopolysaccharide (LPS) was investigated by Northern blot analysis, enzyme immunoassay, and bioassay. The IL-6 mRNA expression began to increase after 1 hr and continued after up to 8 hr of exposure on stimulation with 10 microg/ml of P. intermedia LPS. The bioactivity was dose-dependent on the concentration of P. intermedia LPS (0 to 100 microg/ml). The IL-6 mRNA expression was inhibited by actinomysin D and super induced by cycloheximide. Anti-CD14 monoclonal antibody (MY4) inhibited the IL-6 mRNA expression when administered at a 0.5 microg/ml concentration before stimulation with P. intermedia LPS at 1 microg/ml. The immunoregulatory cytokines (interferon-gamma, IL-10, and IL-4) inhibited LPS-induced IL-6 production with a combined treatment. These results suggest the IL-6 expression by pulp cell cultures is CD14-dependent and regulated at the transcriptional level, and a combined treatment with immunoregulatory cytokines may be effective for control of pulpal inflammation due to P. intermedia LPS. PMID- 11485267 TI - Bactericidal and cytotoxic effects of sodium hypochlorite and sodium dichloroisocyanurate solutions in vitro. AB - The antimicrobial and cytotoxic effects of sodium hypochlorite (NaOCl) and sodium dichloroisocyanurate (NaDCC) were evaluated and compared in vitro. The minimal inhibitory concentration and minimal bactericidal concentration of NaOCl and NaDCC were tested for Streptococcus sobrinus, Streptococcus salivarius, Enterococcus faecalis, and Streptococcus mutans. The cytotoxic effect was assessed by using human fibroblast tissue culture. Survival rate was assessed by a protein determination method. Results showed that the minimal inhibitory concentration and minimal bactericidal concentration values of NaOCl and NaDCC for the tested bacteria were in a similar range. NaDCC in concentrations higher than 0.02%, and NaOCl in concentrations higher than 0.01% were lethal to fibroblasts. In conclusion it seems that both agents were very effective in killing bacteria, and their cytotoxicity to fibroblasts in tissue culture was similar. PMID- 11485268 TI - Mineral trioxide aggregate repair of lateral root perforations. AB - This study was conducted to observe the healing process of intentional lateral root perforation repaired with mineral trioxide aggregate (MTA). Forty-eight root canals of dogs' teeth were instrumented and filled. After partial removal of the filling, an intentional perforation was made with a bur in the lateral area of the root. The perforations were repaired with MTA or Sealapex (control group). Histological analysis occurred 30 and 180 days after treatment. Results showed no inflammation and deposition of cementum over MTA in the majority of the specimens. In the 180-day period, Sealapex exhibited chronic inflammation in all the specimens and slight deposition of cementum over the material in only three cases. In conclusion, MTA exhibited better results than the control group. PMID- 11485269 TI - Analysis of arylsulfatases A and B, acid phosphatase, lactate dehydrogenase, and aspartate transaminase in chronic periapical lesions of endodontic origin. AB - Attempts were made to detect and measure the activities of arylsulfatases. A&B acid phosphatase, lactate dehydrogenase, and glutamate oxaloacetate transaminase (aspartate transaminase) enzymes in human chronic lesions of endodontic origin. Thirteen periapical lesions of endodontic origin and 11 noninflamed control periapical tissues were obtained. The specimens were carried to the laboratory on liquid nitrogen and kept at -70 degrees C. Samples were thawed, homogenized, and then assayed for enzyme activities. The specific activities of arylsulfatase A (nmol/hr/mg protein) were 55.0+/-10.7 (chronic lesions) vs. 3.4+/-2.2 (controls) (p < 0.01). Arylsulfatase B specific activities (nmol/hr/mg protein) were 50.3+/ 6.4 (chronic lesions) vs 91.8+/-18.4 (controls). Total acid phosphatase activities (mU/mg protein) were 45.8+/-6.6 (chronic lesions) vs. 26.8+/-3.1 (controls). Lactate dehydrogenase activities (Berger-Broida units/mg protein) of the chronic periapical lesions were significantly higher than the control group (362+/-63.2) vs. (140+/-46.0) (p < 0.05). There was no significant difference between the specific activities of aspartate transaminase in chronic lesions and the control group (68.0+/-14.5) vs. (53.0+/-10.4) mU/mg protein). PMID- 11485270 TI - Improving the seal of amalgam cores with cemented dowels: a comparative in vitro radioactive tracer study. AB - Amalgam cores with cemented dowels are commonly used to restore endodontically treated teeth. These restorations were widely studied for their mechanical properties; however, less is known about their sealing ability. The effect of the cement type used for the dowel and that of a bonding agent application were studied in a radioactive tracer coronal leakage study. The sealing ability of posts and cores was also compared with that of 5 mm remaining root canal fillings. Composite luting cement provided a better seal than zinc phosphate cement. The seal provided by 5 mm root canal fillings was poorer than that provided by posts and cores. High variability in seal quality was found among all types of posts and cores studied. This may indicate that the seal provided by cemented dowels with amalgam cores is unpredictable. Because neither the remaining root canal filling nor the post and core may be trusted alone for a seal, each should be performed with the greatest care and both covered with a crown as soon as possible. PMID- 11485271 TI - The unpredictability of seal after post space preparation: a fluid transport study. AB - A root canal filling remaining after post space preparation is commonly expected to provide adequate seal. Coronal leakage of 30 endodontically treated teeth was measured before post space preparation using a fluid transport assay. In 10 of these teeth post space was prepared, using a two-step procedure, first to a remaining filling of 6 mm and then to 3 mm, with the leakage studied after each step. In 10 teeth the removal was done in one step to a remaining length of 3 mm. The other 10 teeth, with intact root canal fillings, served as controls and were tested twice for leakage. A significant difference was found between the sealing ability of intact fillings and that of partially removed ones (p < 0.05). The difference between the sealing ability of 3 and 6 mm remaining length group was not statistically significant. The lack of statistical differences between the 6 mm and 3 mm fillings was due to a great variability which existed among the 3 mm remaining fillings. These results suggest that 3 to 6 mm fillings provided a seal inferior to that of intact root canal fillings. Reduction of the fillings to 3 mm resulted in an unpredictable seal. PMID- 11485272 TI - Radiographic measurement of residual root thickness in premolars with post preparation. AB - A minimum thickness of 1 mm has been suggested for the residual root wall in post preparation. The objective of this paper is to evaluate the reliability of radiography when used to measure root thickness. In 106 upper first premolars with one and two root canals, post preparations were made with Peeso reamers to a depth in the root equal to the crown length. Bucco-palatal radiographs were taken of each tooth. The films were processed and projected with a magnification of x20. The smallest thickness of the mesial and distal walls of each root at the apical end of the preparation was measured and recorded. The teeth were then cut at the level of the measurement, and the smallest thickness of the proximal walls was measured using a microscope with a micrometric eyepiece. Fourteen percent of the teeth with one root canal and 27% of the teeth with two root canals could not be measured due to blurred radiographic contours. Variance analysis showed a highly significant difference (p < 0.001) when radiographic and anatomical measurements were compared. The radiograph showed greater thicknesses than were actually present and should not therefore be considered to be a reliable method for measuring residual thickness of tooth walls after post preparation. PMID- 11485273 TI - Relationship between design features of endodontic instruments and their properties. Part 3. Resistance to bending and fracture. AB - Stainless-steel prototypes characterized by five different cross-sectional shapes (square, triangular, rhomboidal, "S"-shaped, and the cross-sectional design of H type files) and three different number of flutes (16, 24, and 32) were used for investigation of the relationship between design features and the resistance to bending and fracture of root canal instruments. Both resistance to bending (bending moment) and resistance to fracture (angular deflection and torque) were determined in accordance to ISO 3630-1. Numbers 15, 25, and 35 prototypes were tested with a sample size of 10 instruments each. Whereas prototypes with the rhombus-shaped cross-sectional design displayed the lowest resistance to bending, those of the square cross-section showed the greatest. In general, S-shaped prototypes and those with the H-type cross-section achieved lower angular deflection than all other prototypes, whereas those with the triangular cross sectional shape and 32 flutes reached the greatest angular deflection. Overall, rhombus-shaped prototypes showed lower torque values than all other prototypes. Results indicate that bending and torsional properties of endodontic instruments are mainly influenced by their cross-sectional design. Moreover, these properties are also affected by the number of flutes and the manufacturing process (twisted or milled) of endodontic instruments. PMID- 11485274 TI - Stafne's bone cavity in the anterior mandible: a possible diagnostic challenge. AB - Stafne was the first to report the presence of "bone cavities" in the angle of 35 mandibles. Such cavities generally appear in the area between the mandibular first molar and the mandibular angle, and are not considered rare. One of their primary radiological diagnostic features is the characteristic location below the mandibular canal. Stafne's bone cavity is relatively rare in the anterior mandible. The mandibular canal is not present in the anterior mandible. As a result, diagnosis in the anterior mandible may be missed. Needless treatment modalities such as endodontic treatment, bone trephining, and bone exploration may be conducted. The purpose of the present report is to describe a new case of Stafne's bone cavity in the anterior mandible and discuss the differential diagnosis process. The dental computerized tomography scan is suggested as the most suitable noninvasive diagnostic and follow-up modality for this bony configuration in the anterior mandible. PMID- 11485275 TI - Heart rate variability in chronic heart failure. AB - The analysis of heart rate variability (HRV) provides information about autonomic cardiovascular control in healthy subjects. In the past 15 years, several articles have been published regarding HRV and chronic heart failure (CHF). The results of these papers substantially demonstrated that HRV is significantly different in CHF patients compared to controls. Moreover, some variables derived from HRV analysis showed significant independent prognostic capacity. In particular, the reduction of variance (expressed as SDNN) and low-frequency spectral component of HRV (ranging from 0.03 to 0.15 Hz) seem related to an increased mortality in CHF. Nevertheless, these variables are not yet considered in clinical practice. A better understanding of the physiopathological basis of the reported alterations of HRV in CHF patients is required in order to permit its use as a clinical tool for prognosis and tailored therapy in individual CHF patients. PMID- 11485276 TI - Non-linear dynamics of cardiovascular system in humans exposed to repetitive apneas modeling obstructive sleep apnea: aggregated time series data analysis. AB - OBJECTIVE: Testing the hypothesis that alterations in non-linear dynamics of the cardiovascular variability appear in healthy, awake subjects during voluntary apnea. SUBJECTS AND METHODS: Ten young subjects performed 20 apneas 60 s each separated by 1 min free breathing. Inter-beat interval (IBI) measured as RR interval in ECG, systolic (SYS) and diastolic (DIAS) arterial blood pressure. stroke volume (SV), cardiac output (CO) and total peripheral resistance (TPR) were non-invasively recorded and computed by Portapress 2 system. Correlation dimension according to Grassberger-Proccacia algorithm (CD) and ratio of % determinism versus % recurrence (RDR) computed from recurrence plot according to Webber and Zbiluth [J. Appl. Physiol. 76 (1994) 965] were used as the indices of chaos and complexity. Sequential time series of cardiovascular variables in consecutive 60-s apneas and inter-apnea free breathings were separately windowed and aggregated. CD and RDR of aggregated apneic time series were compared with CD and RDR values of aggregated time series of inter-apnea free breathing, 10 min at rest and 10 min recovery. Reliability of the aggregation method of sequential time series was tested on transformed simulated data generated by Lorenz model. Error in CD and RDR estimation did not exceed 5% and 1%. respectively. RESULTS: CD of cardiovascular variables computed from aggregated apneas was significantly reduced and RDR augmented versus control and recovery periods. CD and RDR values of inter-apnea phases were in between those in control and during apneas. Time domain linear variance (SD) was increased during aggregated apneas. CONCLUSIONS: Signal dynamics dependent windowing and data aggregation could be a useful tool for non-linear analysis of short repeatable sequential time series; time domain linear variability of the cardiovascular dynamics is augmented while complexity reduced during apneic chemoreceptor stimulation; alterations in non-linear dynamics of cardiovascular variables during apneas in healthy normotensive subjects may suggest an early sign of a mechanism implicated in arterial hypertension in OSAS. PMID- 11485277 TI - Effects of drugs on the autonomic control of short-term heart rate variability. AB - The autonomic nervous system links the brain and the heart. Efferent links in the neural control of the heart consist of sympathetic and parasympathetic (vagal) fibers innervating the sinus node. Because sympathetic and vagal firing alter spontaneous sinus node depolarization, cardiac rate and rhythm convey information about autonomic influences on the heart. The easy availability of ECG rendered possible the assessment of sinus rhythm as an index of autonomic outflow. The frequency-domain approach uses non-invasive recordings and appears to provide a quantitative evaluation of the autonomic modulation of cardiovascular function. Spectral profiles resulting from vagal or sympathetic blockades at the cardiac (or vascular) level might be used as references to unravel the mechanism of action of the drug under examination. A more comprehensive assessment will be obtained if spectral analysis is used as a complement to existing techniques applied for describing the neurohumoral status of patients (microneurographic recordings, norepinephrine spillover). This review also reports some pitfalls encountered in variability studies. PMID- 11485278 TI - Genetic factors contribute to the variance in frequency domain measures of heart rate variability. AB - Power spectral analysis of heart rate variability (HRV) provides quantitative phenotypic markers of autonomic nervous system activity. Reported determinants of HRV only partially explain its variability in the population. The purpose of this study was to estimate the contribution of genetic factors to the variance in HRV measures and assess the heritability of HRV. Subjects who underwent Holter recordings at a routine examination were eligible, excluding subjects with congestive heart failure, coronary artery disease, diabetes mellitus and those taking cardioactive medications. We analyzed the low-frequency power (LF), high frequency power (HF), LF/HF ratio, very low-frequency power (VLF) and total power (TP). Heritability analysis was done by studying correlations between siblings (n = 682, in 291 sibships, 517 pairs) and between spouse pairs (n = 206 pairs). Adjustments were made for sex, age, systolic and diastolic blood pressure, heart rate, coffee and alcohol intake. SAS procedure MIXED was used to estimate and test significance of correlation within sibling pairs and within spouse pairs. Results from separate models were combined to estimate the components of variance of each phenotype, i.e. variance attributable to measured covariates, additive genetic effects (heritability) and household effects. After adjusting for covariates, the correlations were consistently higher among siblings (0.21-0.26) compared to spouses (0.01-0.19). The measured covariates in general accounted for 13-40% of the total phenotypic variance, whereas genes accounted for 13-23% of the variation among HRV measures. Genetic factors contribute towards a substantial proportion of the variance in heart rate and HRV. Recognition of the genetic determinants of HRV may provide additional insight into the pathophysiology of the autonomic nervous system and offer clues toward its modulation. PMID- 11485279 TI - Absence of slowest oscillations in short term heart rate variability of post myocardial infarction patients. GISSI-3 arrhythmias substudy. GISSI-3 Arrhythmias Substudy Investigators. AB - Despite the widely demonstrated association of reduced heart rate variability (HRV) to bad prognosis after myocardial infarction (MI), reference values for HRV parameters are still not available. The GISSI-3 Arrhythmias Substudy studied short-term HRV in a relatively unselected population of patients (324) with recent MI (13 +/- 7 days) providing the statistical description of the main time and frequency domain parameters. All HRV indices, except for the RR interval, showed a non-normal distribution generally skewed around the lowest values. Particularly, no LF power was detected in 75 patients (23%) by power spectral analysis. The absence of LF oscillation in RR spectra was associated to the lower standard deviation of normal RR intervals (SD), aging (> 65 years) and blood pressure hypertension. This result seems to indicate a paradoxical effect of sympathetic overactivity in post-MI patients. PMID- 11485280 TI - Estimation of autonomic response based on individually determined time axis. AB - The analysis of the time-dependence of autonomic response requires: 1. A reliable procedure for the quantification of autonomic activity under nonsteady conditions, such as an algorithm for time-frequency decomposition (ex. SDA. Wigner-Ville, or others). 2. The choice of an adequate time scale for focusing on the data: (a) the regular, universal time scale, independent of the unsteady physiological conditions, or (b) a time axis defined by specific events related to an applied perturbation, as the indicators of specific experimental or physiological conditions, so that each individual is considered according to his own intrinsic time scale. The alignment of the various subjects according to their intrinsic time scale, reflecting their individual response mechanisms, may help to disclose a common pattern of autonomic function. Using an absolute time scale to align and average results for different subjects may obscure the underlying mechanisms. Several examples of autonomic challenges are presented, in which the use of an individual time scale contributes to unveil a typical response pattern: tilt test in vasovagal syncope, the autonomic effect of active standing on hypertension, and the autonomic response to acute hypoxia. PMID- 11485281 TI - Bimodal dose-dependent effect on autonomic, cardiac control after oral administration of Atropa belladonna. AB - This single-blind placebo-controlled study was designed to investigate the dose dependent vagolytic and vagotonic effects after a single oral administration of Atropa belladonna tincture (ABT, 0.1 mg/ml alkaloid concentration, atropine/scopolamine = 20:1). In eight healthy young subjects, heart rate and noninvasive arterial finger blood pressure were recorded simultaneously over 4 h after oral application of four different doses of ABT (day 1: 2 ml, day 2: placebo, day 3: 5 ml, day 4: 1 ml). On each day, 14 20-min sequences under controlled experimental conditions were performed. Among others, mean RR interval (RR), high-frequency spectral power of heart rate variability (HF), and noninvasive baroreflex sensitivity (BRS) were calculated during metronome breathing in supine position. These parameters were robust markers of vagal activity. One hour after 5ml ABT, RR, HF and BRS decreased clearly in six of eight subjects. This effect was interpreted as vagolytic response. After 1 and 2 ml ABT, and after placebo, RR and HF increased markedly. The increase after ABT was much higher than the increase solely due to adaptation after placebo administration, and it could be clearly identified as an augmentation of vagal cardiac activity caused by low-dose ABT. In conclusion, low doses of orally administered ABT can be effectively used to stimulate parasympathetic activity in man. The mode of vagal activation changes between 2 and 5 ml ABT from vagotonic to vagolytic. ABT has no or very little effect on blood pressure control. PMID- 11485282 TI - Wavelet decomposition analysis of heart rate variability in aerobic athletes. AB - Heart rate variability (HRV) can be quantified, among others, in the frequency domain using digital signal processing (DSP) techniques. The wavelet transform is an alternative tool for the analysis of non-stationary signals. The implementation of perfect reconstruction digital filter banks leads to multi resolution wavelet analysis. Software was developed in LabVIEW. In this study, the average power was compared at each decomposition level of a tachogram, containing the consecutive RR-intervals of two groups of subjects: aerobic athletes and a control group. Compared to the controls, the aerobic athletes showed an increased power in all frequency bands. These results are in accordance with values obtained by spectral analysis using the Fourier transform, suggesting that wavelet analysis could be an appropriate tool to evaluate oscillating components in HRV, but in addition to classic methods, it also gives a time resolution. PMID- 11485283 TI - Analysis of heart rate variability with correlation dimension method in a normal population and in heart transplant patients. AB - INTRODUCTION: The correlation dimension (CD) method is related to chaos theory and is used to quantify heart rate variability (HRV). The CD is a measure for the amount of correlations present in the signal. METHODS: The algorithm used to calculate the CD is based on the method of Grassberger and Proccacia. The method was first validated on signals with known CD and then applied to HRV-signals of heart transplants and an age-matched control group of healthy subjects. The CD of the corresponding surrogate time series was calculated to investigate non-linear correlations in the HRV-signal. Circadian variations of the CD were studied in 20 healthy subjects, including men and women. RESULTS: The value of the CD for healthy subjects ranged from 2.12 to 5.53 with a mean value of 4.32. For heart transplants, only a few time series showed a finite value of the CD that varied between 2.10 and 5.60. Also, a significant difference was found between the CD of the original and the surrogate time series in healthy subjects. The CD of women is higher than the CD of men, and this difference was more pronounced during the night than during the day. CONCLUSIONS: This limited study shows that the CD alone cannot be used to make a distinction between HRV-signals of healthy subjects and of heart transplants. However, there is evidence that there are non linear correlations present in the HRV-signal and that there are significant gender and circadian differences in the CD. PMID- 11485284 TI - Phentolamine suppresses the increase in arteriolar vasomotion frequency due to systemic hypoxia in hamster skeletal muscle microcirculation. AB - Systemic hypoxia (8%, 11% and 15% oxygen gas mixture inspiration) has been shown to increase the frequency of arteriolar rhythmic diameter changes in hamster skeletal muscle microcirculation. The effects of phentolamine on vasomotion frequency during systemic hypoxia were studied in Syrian hamsters implanted with a plastic chamber in the dorsum skin. Phentolamine (50 microg/100 g body wt.) was injected intravenously before the 20-min exposure to 11% oxygen gas mixture. The microvessels were studied with a fluorescent microscopy technique, using fluorescein isothiocyanate bound to dextran (mol. wt. 150,000) as a tracer. Vessel diameters were measured with a shearing method. Fourier transform and autoregressive modeling were used to assess the time variant features of diameter changes. Under baseline conditions, the arterioles were characterized by rhythmic diameter changes with fundamental frequency related to vessel size. The terminal branchings were dominated by order 3 vessel activity (frequency: 0.08-0.16 Hz) spreading downstream to all daughter arterioles. Systemic hypoxia caused an increase in vasomotion frequency of order 3 arterioles up to 0.3-0.5 Hz (average: 0.40 +/- 0.06 Hz) and a significant decrease in mean diameter (-28 +/- 5%). Phentolamine completely suppressed the rhythmic changes in diameter of order 3 arterioles that dilated significantly (+ 30 +/- 4%). Therefore, the effects of systemic hypoxia on arteriolar vasomotion appear to be triggered by an increase in sympathetic nervous discharge that induces a rise in frequency up to 0.3-0.5 Hz. PMID- 11485285 TI - Loss of nocturnal dipping of blood pressure and heart rate in obesity-induced hypertension in rabbits. AB - We have investigated in rabbits whether overfeeding and weight gain, which lead to hypertension, are associated with changes in circadian rhythm of blood pressure (BP) and heart rate, and whether the sympathetic nervous system is involved in these changes. In adult male rabbits, mean arterial pressure (MAP) and heart rate (HR) were monitored by telemetry 22 h a day. Daily MAP and HR records were divided into four equal intervals and used to calculate day-night differences. After a 1-week control period, animals were switched to a high-fat (HFD) ad libitum diet for 8 weeks. HFD increased whole day MAP and HR, and rapidly abolished the normal diurnal rhythm of MAP and HR. Since HFD abolished the nocturnal dip in MAP, but had little effect on daytime values, the loss of dipping appears to account for most of the hypertension in this model of obesity. In a separate set of rabbits, alpha- and beta-adrenergic blockade (terazosin + propranolol) prevented HFD-induced hypertension and attenuated the increase in HR by more than half. Adrenergic blockade alone abolished the diurnal rhythm of MAP, chiefly by preventing daytime elevation of MAP. The addition of HFD ad libitum did not further modify daily MAP or its circadian pattern. The diurnal rhythm of HR was relatively unaffected by alpha + beta blockade alone, but was abolished after switching to HFD. In conclusion, rabbits fed an HFD ad libitum develop hypertension and tachycardia associated with a loss of the normal diurnal rhythm of MAP and HR. The hypertension appears to be sympathetically mediated. PMID- 11485286 TI - Nonlinear parameters estimation from sequential short time data series. AB - Procedures of nonlinear parameter estimation require large samples of data. In stationary physiological situations, usually short time series are available. The method of dynamics-dependent windowing and data aggregation procedure are proposed. This technique was tested on chaotic signal generated by Lorenz model and applied to investigate beat-to-beat control of the cardiovascular system in 10 healthy volunteers. Nonivasively recorded blood pressure, respiratory activity and blood oxygen saturation were digitized and saved for further off-line analysis. The experimental procedure consisted of 10 min control--C, 20 voluntary apneas 1 min each-A, interapnea 20 periods of 1 min spontaneous breathing--B, and 10 min free-breathing recovery--R. Respiration signal served as a reference for apnea and interapnea free-breathing identification period. Correlation dimension CD, according to Grassberger and Procaccia, and recurrence plot strategy, according to Webber and Zbilut, were applied to check dynamical properties of the signals. Results of numerical experiment on Lorenz model, original and transformed by segmentation and aggregation, support our assumption of similarity of their dynamics. Error in CD and recurrence parameters estimation strongly depended on segment length and was about 5% for 600 to 1,200 data points. However, even for segments of 75 to 100 samples, it did not exceed 10% for all, but one, periodic testing signal. Segmentation and aggregation applied to interbeat interval (IBI) and total peripheral resistance (TPR) data showed that CD and recurrence variables estimated separately for apneic and interapneic period and those calculated for mixed (apneic and interapneic) intervals were different. Average CD and recurrence parameters of IBI and TPR for 10 subjects during apnea and interapnea intervals were significantly different than during control and recovery. The lowest CD (mean +/- S.D.) of 6.38 +/- 0.4, 5.62 +/- 0.2 and %recurrence 10.35 +/- 0.8, 6.62 +/- 0.6 (highest ratio 4.95 +/- 0.2, 5.13 +/- 0.3) were observed in apnea for IBI and TPR, respectively. Low values of the estimates computed for mixed periods may suggest the influence of slowly varying, quasiperiodic driving force due to experimental procedure regime. Signal dynamics dependent windowing and data aggregation regardless of the sequence of data could be a practical solution for nonlinear analysis of very short repeatable time series. PMID- 11485287 TI - Use of time frequency analysis to follow transitory modulation of the cardiac autonomic system in clinical studies. AB - Heart rate variability (HRV) can be assessed through a time frequency analysis, the smoothed pseudo Wigner-Ville transformation (SPWVT). Such an analysis has allowed the calculation of ICF, a frequential index, extremely sensitive to the modulation of the sympatho-vagal balance. The use of ICF has been applied in the present study to two clinical situations for which a role of this balance was probable, brain death and atrial fibrillation (AF). The use of the time frequency analysis immediately identified the timing of brain death. No difference could be found in the analysis of the 30 min preceding an atrial fibrillation episode. It is suggested that a time frequency analysis should be used in clinical situations in which transitory fluctuations of the sympatho-vagal balance are expected and crucial to the well being of the patients. PMID- 11485288 TI - Sympathetic rhythms and cardiovascular oscillations. AB - Spectral analysis of heart rate and arterial pressure variabilities is a powerful noninvasive tool, which is increasingly used to infer alterations of cardiovascular autonomic regulation in a variety of physiological and pathophysiological conditions, such as hypertension, myocardial infarction and congestive heart failure. A most important methodological issue to properly interpret the results obtained by the spectral analysis of cardiovascular variability signals is represented by the attribution of neurophysiological correlates to these spectral components. In this regard, recent applications of spectral techniques to the evaluation of the oscillatory properties of sympathetic efferent activity in animals, as well as in humans, offer a new approach to a better understanding of the relationship between cardiovascular oscillations and autonomic regulation. PMID- 11485289 TI - Advanced spectral methods for detecting dynamic behaviour. AB - The traditional analysis in the frequency domain of cardiovascular variability signals requires stationarity along the considered temporal window, in order to obtain reliable indicators of the sympatho-vagal balance (low frequency (LF) and high frequency (HF) power and frequency, and LF/HF ratio). Through proper advanced algorithms of signal processing, it is possible to implement methods that allow the enhancement of important parameters about the behaviour of the system under investigation in the time and frequency domain. Both non-parametric and parametric time-frequency methods are generally employed at this purpose. Among them, Wigner-Ville Distribution and Time-Variant Autoregressive models are here described. Through such advanced methods of signal processing, it is possible to investigate the dynamic properties of the spectral parameters during transient physiological or pathological episodes, after a proper validation using simulated signals. The methods are used in various applicative areas of interest where the spectral parameters present a significant change in time and where the classical spectral analysis cannot be correctly applied. A few significant cases will be discussed such as tilting manoeuvre, vaso-vagal syncope onset and progression, and acute ischemic episodes. Further, multivariate analysis can be applied in which the focus is on squared coherence function and phase relationships, in order to estimate some possible causal effects in different experimental conditions. It is believed that such advanced methods of time variant or time-frequency approaches are capable of overcoming the problem of stationarity in classical spectral analysis and to make applicable frequency domain techniques in the study of transient episodes which generally characterise various physiological and clinical conditions. PMID- 11485290 TI - Afferent vagal modulation. Clinical studies of visceral sensory input. AB - The frequency composition of a continuous time series of R-R intervals may be viewed as the phasic output of a central processing system intimately dependent on sensory input from a variety of afferent sources. While different measures of heart rate variability permit a glimpse into the autonomic efferent limb of this complex system, direct access of afferent fibers in humans has remained elusive. Using a specially designed esophageal catheter/manometer probe, we have been able to gain access to vagal afferent fibers in the distal esophagus. Our studies on the effect of vagal afferent electrostimulation on both cerebral evoked potentials (EvP) and the power spectrum of heart rate variability have yielded the following observations: 1. Stimulation of esophageal vagal afferents dramatically and reproducibly increases the high frequency (HF) vagal power and reduces the low frequency (LF) power of the heart rate autospectrum. 2. This effect is constant across stimulation frequencies from 0.1 to 1.0 Hz and across stimulation intensities from 2.5 to 20 mA. 3. Regardless of the stimulation parameters, there are only minimal changes in heart rate (2-6 bpm) and no change in respiratory frequency. 4. There is a linear correlation between electrical stimulation intensity and the amplitude of cerebral evoked potentials, whereas there is a non-linear relationship with all short-term power spectral indices. 5. While cerebral evoked potentials are only elicited at stimulation intensities above perception threshold, there is already a significant shift to increased vagal efferent modulation well below perception threshold. CONCLUSION: These studies support the concept that power spectral indices of heart rate variability represent phasic output responses to tonic afferent viscerosensory signals in humans. These studies also demonstrate the feasibility of accessing vagal afferents in humans. PMID- 11485291 TI - Rhythms, rhymes, and reasons--spectral oscillations in neural cardiovascular control. AB - Cardiovascular neural regulation is an integrated response to a continuous interaction of inhibitory and excitatory stimuli. Neural control of the circulation appears to be coded simultaneously in different modalities as amplitude (strength of signal or tonic activity) and frequency (oscillatory or phasic activity). Changes in tonic activity appear to be accompanied by tightly linked modulations in oscillatory characteristics. This is true within a narrow range of physiologic conditions, and the relationship is eliminated in extreme cardiovascular pathophysiology. Nevertheless, the oscillatory patterns in cardiovascular neural control appear to be widespread so that low and high frequency oscillatory patterns are evident even in sympathetic traffic to skin (Cogliati et al., 2000). Thus, it is likely that there is a functional significance to these oscillations. Recent data from Nafz et al. (1999) suggest that the presence of LF oscillatory characteristics in renal perfusion may attenuate renin-angiotensin activation during renal hypotension. These findings may have direct relevance to poorer outcomes observed in heart failure patients in whom an absence of LF oscillatory power was observed in RR interval and sympathetic traffic (Van de Borne et al., 1997a). PMID- 11485292 TI - Modulatory effects of respiration. AB - Respiration is a powerful modulator of heart rate variability, and of baro- and chemoreflex sensitivity. Abnormal respiratory modulation of heart rate is often an early sign of autonomic dysfunction in a number of diseases. In addition, increase in venous return due to respiration may help in maintaining blood pressure during standing in critical situations. This review examines the possibility that manipulation of breathing pattern may provide beneficial effects in terms not only of ventilatory efficiency, but also of cardiovascular and respiratory control in physiologic and pathologic conditions, such as chronic heart failure. This opens a new area of future research in the better management of patients with cardiovascular autonomic dysfunction. PMID- 11485293 TI - RR-arterial pressure variability relationships. AB - Methodological aspects of a causal black-box model of heart period/arterial pressure interaction, arterial pressure closed-loop regulation and respiration effects on both heart period and arterial pressure are revisited in the "time" (more exactly heart beat count) domain. Parameters are estimated from experimental data (model identification) by means of multiple linear regressions of actual samples over the past ones. The elements composing either heart period or systolic arterial pressure variability are visualised as beat-by-beat series. Indexes describing the signal interactions, the loop properties and the spectral components of the variability series are consequently summarised. In 17 normal young volunteers, the analysis was carried out during active standing, rest, mild clinostatic pedalling exercise at 10%, 20%, and 30% of the maximum effort, and recovery. A negative effect of heart period changes on systolic arterial pressure of - 13.3 mm Hg/s was found at rest. This effect, though augmented by exercise, appeared insignificant in explaining arterial pressure variability. Arterial baroreflex was assessed by alphaart index which had a value of 5.18 mm Hg/ms at rest, 3.78 mm Hg/ms during active standing, and decreased progressively with exercise down to 0.55 mmHg/ms. The pressure regulation loop displayed a tendency to amplify disturbances at low frequency (around 0.1 Hz) 5.94 times at rest, augmented to 8.88 times during standing, 7.55 at 30% exercise. The first parameter of the pressure auto-regression was slightly higher than 1 at rest and even more augmented during standing, thus, indicating a tendency of arterial pressure perturbations to persist from one beat to the next. These mechanisms appear important in the genesis of low-frequency pressure waves. Nonetheless, the trace of different sources was evident in the regression residuals. Noticeably, during exercise it explained 10.16% of total heart period variability compared to 12.49% related to the low-frequency oscillations of closed-loops. The origin of high-frequency waves synchronous with respiration appeared miscellaneous as well. Arterial pressure appeared negligibly affected by heart period changes. Conversely, a limited effect of arterial pressure waves was found on heart period superimposed to a large effect of cardiopulmonary reflexes directly modulating the sinus node. In conclusion, both high-frequency and low-frequency waves are composite phenomena and a multi-channel analysis comparing heart period and arterial pressure variability yields a variety of figures assessing cardiovascular regulation and cardiorespiratory coupling. PMID- 11485294 TI - Neural mechanisms of cardiovascular regulation during exercise. AB - This brief review addresses current hypotheses concerning the reflex control of circulation during exercise in humans. In particular, the specific objective of this review is to describe how time and frequency domain analysis of blood pressure and heart rate variability signals permitted to gain new insights onto reflex mechanisms of cardiovascular regulation during exercise, without the need of perturbing the cardiovascular system from the outside, utilizing fully noninvasive approaches and avoiding artificially isolating the influence of the different neural pathways involved in the control of the cardiovascular system. Throughout the article, particular emphasis is given to the complexity and plasticity of the neural control of the circulation during exercise, by presenting data that show how the reflex mechanisms involved in cardiovascular regulation, namely, the arterial baroreflex and the muscle metaboreflex, may be differently modulated in relation to the muscular activity being performed, such as the type of exercise, the intensity of exercise and the size of active muscle masses. PMID- 11485295 TI - Autonomic dysregulation in essential hypertension: insight from heart rate and arterial pressure variability. AB - Essential hypertension is the most prevalent cardiovascular disorder, affecting more than 50 million people in the USA. Hypertension-related mortality and morbidity figures have been greatly improved over the last three decades by major advances in prevention. Detection and operative suggestions for practicing physicians are available from several guideline treatments derived from grouped data obtained in numerous well-conducted studies on large numbers of patients. However, the disappointing results of some forms of antihypertensive therapies, particularly in preventing coronary artery disease, has shed some doubts on traditional approaches to managing hypertensive patients. At present, in spite of extensive investigations, the exact causal mechanism(s) are far from being fully understood, and consequently, essential hypertension is still managed using a heuristic approach. Persistent elevations in arterial pressure imply some disturbances in the complex and multifactorial cardiovascular control mechanisms. In this context, neurohumoral disturbances might play a special role, in view of the demonstration that an elevated sympathetic drive seems essential in hypertensive patients. In this review, we follow the hypothesis that other allied methods capable of quantitatively assessing some aspects of the regulatory system might support and integrate the usual dichotomous diagnostic procedure based on arterial pressure determinations. In prior studies, we reported that parameters obtained by spectral analysis of heart rate variability (HRV) might furnish useful information on autonomic normal and abnormal nervous system regulation. In the foregoing, we summarize our experience using this approach in the clinical management of hypertensive patients. It is our tenet that spectral analysis of mono or multivariate cardiovascular beat-by-beat variabilities provides potentially important information on alterations in neural control of the circulation accompanying essential hypertension. In spite of an ongoing debate on the interpretation of specific aspects of HRV spectral components, overall, it appears that the available evidence supports the hypothesis that in essential hypertension, there is an increased sympathetic and reduced vagal cardiac drive coupled with an enhancement of vasomotor sympathetic modulation. Prospective studies on large populations, rendered more easy to perform, thanks to improvements in technology and telemedicine applications, might provide an answer to the still open question of how to apply spectral analysis of HRV to a better mechanistic understanding of essential hypertension, and to more satisfactory individually tailored antihypertensive treatments. PMID- 11485296 TI - Orthostatic intolerance: different abnormalities in the neural sympathetic response to a gravitational stimulus. AB - In this paper we shall focus on the different abnormalities in the neural sympathetic response to a gravitational stimulus, characterising syndromes with symptoms of orthostatic intolerance. In Vaso vagal Syncope, an increase or a reduction of cardiac and vascular sympathetic modulation have been described in occasional and habitual fainters, respectively. Pure Autonomic Failure (PAF) is characterized by a global cardiovascular denervation. Accordingly, the spectral markers of cardiac and vascular sympathetic modulation are absent or reduced. However, a concomitant vagal diminished activity is present. In Chronic Orthostatic Intolerance (COI), the most common form of dysautonomia in young female, an abnormal regional distribution of sympathetic discharge has been hypothesized during standing. Indeed, an overall increased sympathetic activity is present in recumbent position; during tilt a blunted vascular sympathetic discharge, with a concomitant exaggerated cardiac sympathetic modulation, is evident. Baroreflex Failure is a syndrome that may result from neck surgery or irradiation due to different forms of regional cancer. It is characterized by a volatility of blood pressure and heart rate, without habitual orthostatic hypotension. In the present paper, we describe a case of Baroreflex Failure with marked orthostatic hypotension in spite of a huge muscle sympathetic nerve activity (MSNA) and high levels of plasma cathecolamines. The most relevant finding was the absence of any coordinate rythmicity in blood pressure, heart rate and MSNA, both at rest and during tilt, particularly in the frequency band likely to be related with sympathetic modulation, i.e. at 0.1 Hz. We hypothesize that the absence of 0.1 Hz spontaneous fluctuations might play a role in sustaining orthostatic hypotension. PMID- 11485297 TI - Cardiovascular variability characteristics in obstructive sleep apnea. AB - Patients with obstructive sleep apnea (OSA) are at increased risk for cardiovascular disease. Altered cardiovascular variability is a prognostic indicator for cardiovascular events. This review examines the evidence that OSA is accompanied by alterations in cardiovascular variability. This alteration is evident even in the absence of hypertension, heart failure or other disease states, and may be linked to the severity of OSA. The presence of clear-cut abnormalities in time and frequency-domain measures of blood-pressure and heart rate variability in normotensive OSA patients provides intriguing evidence for the concept of an etiologic interaction between sleep apnea and cardiovascular disease. Mechanisms that could contribute to altered cardiovascular variability in patients with sleep apnea include abnormalities in chemoreflex, baroreflex and endothelial function. PMID- 11485298 TI - Heart rate variability in ischemic heart disease. AB - Major untoward events, such as life-threatening arrhythmias and acute coronary events, have been suggested to be triggered by the activation of the autonomic nervous system in patients with coronary artery disease. Analysis of heart rate variability by conventional time and frequency domain methods, as well as by newer methods derived from nonlinear system theory, has offered a novel approach for studying the abnormalities in cardiovascular neural regulation in ischemic heart disease. Heart rate variability has been shown to be altered among patients with ischemic heart disease as compared to their age-matched controls without the evidence of ischemic heart disease. There are also obvious differences in various measures of heart rate variability between patients with uncomplicated coronary artery disease and those with coronary artery disease with complicated myocardial infarction. Impaired high-frequency oscillations of heart rate is the most prominent feature in patients with uncomplicated coronary artery disease, suggesting mainly an impairment in vagal autonomic regulation. Patients with prior myocardial infarction have a reduced overall heart rate variability, and a specific spectral pattern with a reduced low-frequency spectral component has been observed in patients with prior myocardial infarction and impaired left ventricular function. Recent studies have shown that the new nonlinear measures, particularly fractal analysis methods of heart rate dynamics, can detect subtle changes in heart rate behavior that are not easily detected by traditional analysis methods from ambulatory recordings. Patients with prior myocardial infarction have steeper power-law slope analyzed from the ultra and very low frequency spectral bands, and they also have more random short-term heart rate dynamics analyzed by the detrended fluctuation method. A large body of data indicate that reduced overall heart rate variability is associated with an increased risk of mortality and nonfatal cardiac events in patient with ischemic heart disease. Of particular note, recent studies indicate that fractal analysis methods perform even better than the traditional analysis methods of heart rate variability as predictors of death and the onset of life-threatening arrhythmic events in post-infarction populations. These findings support the notion that heart rate variability analysis methods, such as fractal and complexity measures as well as conventional techniques, give valuable clinical information among patients with ischemic heart disease. PMID- 11485299 TI - Circadian rhythm biochemistry: from protein degradation to sleep and mating. PMID- 11485300 TI - Rapid purification of recombinant anthrax-protective antigen under nondenaturing conditions. AB - Anthrax-protective antigen is the central moiety of the anthrax toxin complex that mediates the entry of the other two toxin components, lethal factor and edema factor into the cells. It is also the main immunogen of the cell-free vaccine against anthrax. However, in addition to PA, the vaccine contains trace amounts of other culture-derived proteins that contribute to the side effects of the vaccine like pain, edema, erythrema, etc. Thus there is a need to develop high-resolution purification methods to purify PA to homogeneity. In this study we have presented a purification strategy for rapid purification of recombinant protective antigen under nondenaturing conditions, which ensures that not only biological activity but also the conformational integrity of immunological epitopes is well-preserved. The protein was purified to homogeneity in a two-step purification procedure that takes just 6 h for completion. Three milligrams of recombinant protective antigen obtained from 1-liter culture was comparable to B. anthracis protective antigen in terms of functional and biological activity. Moreover, the immunogenicity elicited by the purified protein in mice was also studied. The studies reported here are part of continuing research that aims to provide a safe and efficacious alternative to the current vaccine against anthrax. PMID- 11485302 TI - Colon cancer cell adhesion to endothelial E-selectin inhibits detachment of endothelial cells through activation of beta(1)-integrin. AB - Previous studies have implicated a role for E-selectin in carcinoma cell adhesion to vascular endothelium. We examined the role of colon cancer cell adhesion to vascular endothelium via E-selectin using adenoviral vector-mediated transfection in human umbilical vein endothelial cells (HUVECs). We found that the amount of HUVEC detachment from the gelatin matrix 24 h after LS-180 cell adhesion was inhibited only when the HUVECs were transduced with wild-type E-selectin, but not with a cytoplasmic domain truncated mutant E-selectin or the control Lac-Z vector. We also found that the adhesion of LS-180 cells to wild-type E-selectin transduced HUVEC-induced activation of beta(1)-integrin receptors without affecting MMP activity. These results indicate that colon cancer cell adhesion via E-selectin inhibits HUVEC detachment from the monolayer, at least in part by modulating beta(1)-integrin activity in HUVECs. In addition, they indicate the importance of the cytoplasmic domain of E-selectin with this phenomenon. PMID- 11485301 TI - Inhibition of retinal guanylyl cyclase by the RGS9-1 N-terminus. AB - Cyclic GMP plays a key role in retinal phototransduction and its photoreceptor concentration is precisely controlled by the cooperative action of cGMP phosphodiesterase (PDE) and retinal guanylyl cyclase (retGC). However, studies of the relationship between these two systems have focused only on a Ca(2+) mediated, indirect connection. Using a retinal "regulator of G-protein signaling" (RGS9-1) and its fragments, we show that the N-terminus of RGS9-1 inhibits retGC activity. We also indicate that the GGL domain and/or the RGS domain function as an internal suppressor against the N-terminus, suggesting that proteins bound to these domains regulate the inhibitory activity of the N-terminus. Direct interaction of retGC with RGS9-1 and its N-terminus is also proved by immunoprecipitation and an overlay technique. Since RGS9-1 also controls the lifetime of transducin-activated PDE through regulating GTPase activity of transducin, this study strongly suggests that RGS9-1 mediates the direct interaction between PDE and retGC systems, and that this ingenious mechanism plays an important role in tuning of cGMP concentration in photoreceptors. PMID- 11485303 TI - The homolytic and heterolytic fatty acid hydroperoxide lyase-like activities of hematin. AB - Pentenols and pentene dimers are biosynthetized in plants by homolytic fatty acid hydroperoxide lyase (HPL) or HPL-like enzymes. It has been found that these compounds can modify the flavor of olive oil. Reactions between hematin and 13 hydroperoxyoctadecatrienoic acid resulted in the formation of the same compounds via a free radical reaction in which an alkoxyl radical derived from linolenic acid hydroperoxide undergoes a beta-scission. (Z)-3-Hexenal has also been detected as a minor product of the reaction. It is bioconversed from the same substrate in plants by heterolytic HPL. Thanks to the redox cycle of its central iron, hematin has both homolytic and heterolytic HPL-like activities. PMID- 11485304 TI - Interleukin-1 regulates FGF-2 mRNA and localization of FGF-2 protein in human osteoblasts. AB - Interleukin-1 (IL-1) and basic fibroblast growth factor (FGF-2) are potent stimulators of osteoclast formation. However, the role of FGF-2 in the responses to IL-1 in bone has not been reported. We examined the effect of IL-1 on FGF-2 mRNA and protein expression in human osteosarcoma MG-63 osteoblasts, normal human osteoblasts (NHOB), and osteoblasts from osteoarthritic patients (F2 and F13). IL 1 increased FGF-2 mRNA expression in osteoblasts within 1.5 to 3 h. Multiple FGF 2 protein isoforms were expressed in human osteoblasts. Twenty-four hours of treatment of MG-63 and NHOB cells with IL-1 increased the high-molecular weight(HMW, 22/24 kDa) and low-molecular-weight (LMW, 18 kDa) FGF-2 proteins intracellularly. In contrast, IL-1 preferentially increased the LMW protein signal intracellularly as well as on the cell surface of F2 and F13 osteoblasts. We conclude that IL-1 is a major stimulator of FGF-2 expression in human osteoblasts. Furthermore, selective increases in the exportable LMW protein in osteoblasts from osteoarthritic patients may be of clinical relevance. PMID- 11485305 TI - Allosteric regulation of CCR5 by guanine nucleotides and HIV-1 envelope. AB - The chemokine receptor CCR5 is the principal coreceptor for R5 (macrophage tropic) strains of HIV-1. CCR5 uses G-proteins as transducing elements. Here we report the biochemical consequences of the interaction between CCR5 and G proteins. Macrophage inflammatory protein-1beta (MIP-1beta) binding to CCR5 was potently and specifically inhibited by guanine nucleotides. The molecular mechanism of this inhibitory effect was shown to be a dose-dependent reduction in MIP-1beta receptors. We also show that the MIP-1beta binding site is allosterically regulated by monovalent cations and that binding of this endogenous agonist is highly temperature sensitive and dependent on divalent cations, characteristic of a G-protein-coupled receptor(GPCR). HIV-1 envelope glycoprotein decreased the affinity of CCR5 for MIP-1beta but also altered the kinetics of MIP-1beta binding to CCR5, proving that it interacts with a distinct, but allosterically coupled binding site. The findings described herein contribute to our understanding of how CCR5 interacts with chemokines and HIV-1 envelope. PMID- 11485306 TI - Activities and kinetic mechanisms of native and soluble NADPH-cytochrome P450 reductase. AB - Native yeast NADPH-cytochrome P450 oxidoreductase (CPR; EC 1.6.2.4) and a soluble derivative lacking 33 amino acids of the NH(2)-terminus have been overexpressed as recombinant proteins in Escherichia coli. The presence of a hexahistidine sequence at the N-terminus allowed protein purification in a single step using nickel-chelating affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis confirmed the predicted molecular weights of the proteins and indicated a purity of >95%. Protein functionality was demonstrated by cytochrome c reduction and reconstitution of CYP61-mediated sterol Delta(22)-desaturation. Steady-state kinetics of cytochrome c reductase activity revealed a random Bi-Bi mechanism with NADPH donating electrons directly to CPR to produce a reduced intermediary form of the enzyme. The kinetic mechanism studies showed no difference between the two yeast CPRs in mechanism or after reconstitution with CYP61-mediated 22-desaturation, confirming that the retention of the NH(2) terminable membrane anchor is functionally dispensable. PMID- 11485307 TI - Cooperative regulation of CYP3A5 gene transcription by NF-Y and Sp family members. AB - The purpose of this study was to clarify the mechanism(s) responsible for the transcriptional regulation of the human CYP3A5 gene. It was found that a region from nucleotides -90 to -40 was involved in the transcriptional activity of the CYP3A5 gene by transfection of a series of 5'-truncated promoter-luciferase chimeric genes into human hepatoma HepG2 cells. A gel shift assay using nuclear extracts prepared from HepG2 cells showed that nuclear factor-Y (NF-Y) and specificity protein (Sp) 1 and Sp3 bound to CCAAT box (-78/-68) and a basic transcription element (BTE) (-67/-46) in the CYP3A5 gene. Furthermore, introduction of mutations in the CCAAT box, the BTE, or both elements decreased the transcriptional activity to 10, 21, or 4% of that seen with the intact gene. Thus, we conclude that the transcription of the CYP3A5 gene is cooperatively regulated by NF-Y, Sp1, and Sp3 in HepG2 cells. PMID- 11485308 TI - C3G, a guanine nucleotide exchange factor bound to adapter molecule c-Crk, has two alternative splicing forms. AB - Two types of C3G cDNA were isolated from mouse 3T3-L1 adipocyte cDNA library. A 114-bp sequence in the middle of C3G cDNA is deleted in the short type cDNA. By RT-PCR analysis, it was found that these two types of C3G mRNA existed in all the mouse tissues. Sequence comparison revealed 88% nucleotide sequence identity between mouse and human C3G cDNA. Comparison of mouse C3G cDNA with the human genome database suggested that this 114-bp sequence comprised an entire exon, and it is confirmed by PCR analysis using mouse genomic DNA and cDNA template. These results indicate that two C3G mRNAs and proteins result from alternative RNA splicing. PMID- 11485309 TI - Molecular cloning of silkworm paralytic peptide and its developmental regulation. AB - The silkworm paralytic peptide (PP) is a member of the ENF peptide family that exerts multiple biological activities involved in defense reaction and growth regulation. We isolated its cDNA and examined mRNA expression profiles. cDNA encoded 131 amino acids from which the 23-residue PP sequence was found at the C terminal portion. Immunoblot analysis and paralytic activity assay indicated that inactive pro-protein in larval hemolymph was processed into active peptide immediately after bleeding. In the last larval instar, 0.6-kb PP mRNA was expressed in various tissues, of which the fat body was predominant. Its expression in the fat body decreased during the feeding period and then increased during metamorphic process. Juvenile hormone and 20-hydroxyecdysone upregulated its expression. At the embryonic stage, 1.5-kb mRNA, in addition to 0.6-kb mRNA, was expressed from 1 day after oviposition to hatching. PP was thus expressed stage-specifically under hormonal control. PMID- 11485310 TI - Analysis of type-I and type-II RUNX2 protein expression in osteoblasts. AB - Runt-related transcription factor-2 (RUNX2) is expressed as two isoforms (type-I and type-II) differing only in their amino terminal sequences. The amino terminus of type-I contains MRIPV instead of MASNSLFSAVTPCQQSFFW in type-II. Although type II mRNA has been considered osteoblast specific, the RUNX2 protein isoforms expressed in osteoblasts have not yet been identified. Using antisera generated against the two different amino terminal sequences of type-I and type-II RUNX2, we show the expression of both isoforms in cells with the mature osteoblast phenotype (fetal rat calvarial cells, and ROS 17/2.8, SaOS-2 and U2OS osteosarcoma cell lines), but only type-I in partially differentiated osteoblast like cells (the UMR-106 osteosarcoma cell line). Since UMR-106 cells express both type-I and type-II mRNAs, our results suggest that the translation of type-II mRNA is repressed in these cells. No RUNX1 and RUNX3 proteins are detected in any of the osteoblastic cells tested. The antisera we have generated will be useful for studies relating expression of RUNX2 isoforms to control of osteoblast differentiation. PMID- 11485311 TI - Exon/intron organization and transcription units of the human acyl-CoA synthetase 4 gene. AB - Acyl-CoA synthetase 4 (ACS4) is an arachidonate-preferring isozyme of ACS family predominantly expressed in steroidogenic tissues. Isolation and characterization of genomic clones encoding human ACS4 revealed that the genomic organization of the gene. The human ACS4 gene spans approximately 90 kb and consists of 16 exons. Sequence inspection of the 5'-flanking region revealed potential DNA elements including GATAs, p300, AP-4, SRY, CREB and MyoD. A minimal promoter region required for the expression of ACS4 in HeLa S3 cells was determined. The human ACS4 gene was mapped between the STS markers, WI-17685 and CHLC.GATA81B07 on Xq22 23 region. PMID- 11485312 TI - Regulation of protein kinase CKII by direct interaction with the C-terminal region of p47(phox). AB - Protein kinase CKII is a Ser/Thr kinase which is involved in many proliferation related processes in the cell. p47(phox) is a component of the leukocyte NADPH oxidase, which is an important element of host defense against microbial infection. In this study, we demonstrate that a truncated form of the p47(phox) lacking its N-terminal region (p47(phox)/SH3-C), but not a truncated form of the p47(phox) lacking its C-terminal region (p47(phox)/N-SH3), undergoes better phosphorylation by CKII in the presence of arachidonic acid. The yeast two-hybrid test and the glutathione S-transferase (GST) pull-down assay showed that p47(phox) interacts specifically with the regulatory beta subunit (CKIIbeta), but not with the catalytic alpha subunit (CKIIalpha) of the tetrameric CKII holoenzyme. The binding of p47(phox) to CKIIbeta requires the C-terminal region of p47(phox) and is completely abolished by addition of spermine, indicating that a highly basic region in the C-terminal region of p47(phox) contributes to binding to CKIIbeta. In addition, p47(phox) stimulates the catalytic activity of CKII holoenzyme; this stimulation also requires the C-terminal region of p47(phox). Coimmunoprecipitation experiments showed that CKII holoenzyme interacts with p47(phox) in human neutrophils. Taken together, the present data indicate that the C-terminal region of p47(phox) plays a significant role in the arachidonic acid-dependent phosphorylation of p47(phox) by CKII and that the same region of p47(phox) associates directly with CKIIbeta and can modulate the catalytic activity of CKII holoenzyme. PMID- 11485313 TI - Transcriptional activity of the promoter region of rat frizzled-related protein gene. AB - Frizzled-related protein (Frp) is a new family of secreted proteins involved in tumorigenesis and Wnt-signaling pathway. Previous study has shown that rat Frp (rFrp) gene was found to be differentially expressed in Rat 6 fibroblast cell line overexpressing p53(val135) (R6#13-8). The rFrp gene was otherwise silent in normal parental Rat 6 cells. To elucidate the molecular basis of the transcriptional activation of rFrp, we have isolated and analyzed a 2-kilobase pair promoter region of the rFrp gene. Mapping of transcription initiation sites of rFrp showed the existence of multiple initiation sites. Transfection studies of serial deletion constructs in both Rat 6 and CHOK1 cell lines revealed that the region from -202 to -144 contains cis-acting elements essential for the efficient transcription of rFrp. This work provides a transcriptional regulation basis for Frp and gives insight into its implication in tumorigenesis. PMID- 11485314 TI - Autoregulation of mouse BMP-2 gene transcription is directed by the proximal promoter element. AB - Bone morphogenetic protein-2 (BMP-2) stimulates the commitment and differentiation of precursor mesenchymal cells to mature bone. We have isolated and sequenced 2712 base pairs (bp) of the 5' flanking region of mouse BMP-2 gene. Using RNase protection assay we identified two transcription initiation sites within this 2712 bp region of the BMP-2 gene. The distal start site was mapped to -736 bp in relation to the proximal start site (+1). Recombinant BMP-2 preferentially stimulated transcription initiation from the proximal start site. To investigate the mechanism of transcription initiation from these two start sites, we identified two promoter elements upstream of the proximal and distal transcription initiation sites. Transfection of promoter-luciferase reporter constructs into cells of different organs demonstrated differential transcriptional activity of proximal and distal promoters, with highest activity in the osteoblast cell lineage. In osteoblasts, BMP-2 stimulated transcription from the proximal promoter only. Together our data provide the first evidence for the presence of two transcription initiation sites with two upstream promoter elements in mouse BMP-2 gene. Furthermore, we demonstrate for the first time that BMP-2 autoregulates its expression in osteoblasts through the proximal promoter dependent transcriptional mechanism. PMID- 11485315 TI - Involvement of plasmid in degradation of pentachlorophenol by Pseudomonas sp. from a chemostat. AB - Pseudomonas sp. strain IST103 obtained from a stable bacterial consortium was capable of utilizing pentachlorophenol (PCP) as sole carbon and energy source. The consortium was developed by continuous enrichment in a chemostat. The degradation of PCP by bacterial strain proceeded through an oxidative route as indicated by accumulation of tetrachloro-p-hydroquinone and chlorohydroquinone determined by high performance liquid chromatography (HPLC), and chloride molecules released in culture medium. Two different molecular size plasmids, of approximately 80 and 4 kilobase, were found to be responsible for carrying genes for degradation of PCP. This was evidenced by mutants produced by curing of plasmid by treatment of ethidium bromide. The derivatives were not able to utilize PCP, however, transformation of low molecular size plasmid of Pseudomonas sp. strain 103 into E. coli JM109 utilized PCP, indicated a possible involvement of plasmid in degradation of pentachlorophenol. PMID- 11485316 TI - Accumulation of plant galactolipid affects cell morphology of Escherichia coli. AB - Monogalactosyldiacylglycerol (MGDG) is a major constituent of thylakoid membrane in chloroplasts. Therefore, it is considered to have an important role in the maintenance of the complicated structure of the thylakoid membrane. We have succeeded in cloning the enzyme for MGDG synthesis and overexpressed it in Escherichia coli. In this study we analyzed the morphology of the E. coli harboring the gene. The fatty acid composition of its membrane lipids did not differ between the wild type and transformant, except for the appearance of MGDG. However, transformant cells appeared to be elongated. DAPI staining revealed the entire intracellular region of filamentous cells to be stained; therefore, the elongation of the cells is probably due to a defect in cell division. Atomic force microscopy revealed that the transformant had a smooth but scratched surface. It was concluded that the excessive accumulation of a non-bilayer lipid, MGDG, interfered with the translocation of proteins across the plasma membrane, including those for cell division. PMID- 11485317 TI - Identification of a novel spliceoform of inositol polyphosphate 4-phosphatase type Ialpha expressed in human platelets: structure of human inositol polyphosphate 4-phosphatase type I gene. AB - Inositol polyphosphate 4-phosphatases (IP4Ps) are enzymes involved in the regulation of phosphoinositide 3-kinase (PI3K) signaling. IP4Ps catalyze the hydrolysis of the D-4 position phosphoester of the PI3K generated lipid second messenger, phosphatidylinositol 3,4-bisphosphate. Western blot analysis detected the expression of a novel 110 kDa form of IP4P type Ialpha in mouse spleen, heart, lung, and uterus. In addition, the 110 kDa form of IP4P type Ialpha was found to be the major form of this enzyme expressed in human platelets, MEG-01 megakaryocytes and Jurkat T-cells. RT-PCR analysis of MEG-01 megakaryocytes and Jurkat T-cells indicates that the 110-kDa form of IP4P Ialpha is derived from an alternatively spliced mRNA that encodes an additional internal domain of 40 amino acids not present in the two previously described brain IP4P Ialpha spliceoforms. The predicted molecular mass of this spliceoform is 109,968 Da, consistent with its apparent molecular mass estimated by Western blot analysis. The novel domain is proline rich and contains a PEST sequence characteristic of proteins that are rapidly degraded by the calpain family of proteases. Analysis of genomic DNA sequence indicates that the IP4P type I gene consists of 25 exons and that this novel spliceoform is obtained as a result of an unusual type of differential splicing involving the use of an alternative 5'-GU donor splice site during the excision of intron 15. In addition, we show that all three known spliceoforms of IP4P Ialpha result from alternative splicing involving exon 15 and 16 indicating that structural variability in this region of the enzyme may be important for its function. PMID- 11485318 TI - Changing the substrate specificity of cytochrome c peroxidase using directed evolution. AB - Cytochrome c peroxidase (CCP) from Saccharomyces cerevisiae was subjected to directed molecular evolution to generate mutants with increased activity against 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Using a combination of DNA shuffling and saturation mutagenesis, mutants were isolated which possessed more than 20-fold increased activity against ABTS and a 70-fold increased specificity toward ABTS compared to the natural substrate. In contrast, activities against another small organic molecule, guaiacol, were not significantly affected. Mutations at residues Asp224 and Asp217 were responsible for this increase in activity. These two residues are located on the surface of the protein and not in the direct vicinity of the distal cavity of the peroxidase, where small organic substrates are believed to be oxidized. Mutations at position Asp224 also lead to an increased amount of the active holoenzyme expressed in Escherichia coli, favoring the selection of these mutants in the employed colony screen. Possible explanations for the effect of the mutations on the in vitro activity of CCP as well as the increased amount of holoenzyme are discussed. PMID- 11485319 TI - Isolation and characterization of Vsx1, a novel mouse CVC paired-like homeobox gene expressed during embryogenesis and in the retina. AB - Gastrula stage mouse embryo RNA was screened by degenerate RT-PCR to yield a novel paired-like homeobox gene. The open reading frame encoded by the cDNA was most similar to human VSX1. Mouse Vsx1 encodes a protein of 363 amino acid residues that contains a CVC domain that was originally identified as a conserved motif among mouse CHX10, goldfish VSX-1 and C. elegans CEH-10. Linkage analysis showed that mouse Vsx1 mapped to the distal region of chromosome 2. RT-PCR analysis detected mouse Vsx1 transcripts from gastrulation and post-gastrulation stage mouse embryos, suggesting a role for Vsx1 during mouse embryogenesis. Analysis of the eyes of mouse chimeras generated with embryonic stem cells in which a lacZ reporter was targeted to the Vsx1 locus suggested that Vsx1 is expressed in the inner nuclear layer of the retina. PMID- 11485320 TI - Proatherogenic flow conditions initiate endothelial apoptosis via thrombospondin 1 and the integrin-associated protein. AB - Recently it has been shown that vascular endothelial cells (EC) are completely devoid of apoptosis if cultivated under a steady laminar flow and that apoptosis is induced by turning off the flow. An autocrine loop of thrombospondin-1 (TSP-1) and the alpha(v)beta(3) integrin/integrin-associated protein (IAP) complex has been identified as the molecular coupling device between flow and apoptosis. Lack of blood flow is a rare and mostly transient phenomenon whereas irregular flow conditions are permanently present at arterial bifurcations and sites of abnormal vessel morphology. Irregular flow conditions are established here either by the action of a cone-and-plate type flow apparatus generating a uniform turbulent flow or in a flow chamber by insertion of a local hindrance creating a zone of unsteady laminar flow with vortex formation and lowered shear stress. In both cases apoptosis is induced either throughout the entire monolayer or restricted to the locally defined area. Flow disturbance and apoptosis are coupled by the described autocrine loop of TSP-1 and the integrin/IAP receptor complex. In vivo atherosclerotic lesions occur predominantly at sites of flow irregularities, which are thought to be pro-atherogenic. Thus we propose a key role of the identified mechanosensitive apoptosis induction for the initiation of atherosclerosis. PMID- 11485321 TI - Automated mode-of-action detection by metabolic profiling. AB - Rapid classification and identification of the mode-of-action of bioactive compounds applied to plants can be achieved by a robust and easy-to-use metabolic profiling method. This method uses artificial neural network analysis of one dimensional proton NMR spectra of aqueous plant extracts to rapidly classify changes in the total metabolic profile caused by application of crop protection chemicals. PMID- 11485322 TI - Cytoplasmic gamma actin as a Z-disc protein. AB - To investigate the precise localization of cytoplasmic gamma actin in skeletal muscle and the relationship to dystrophin molecules, we designed an antibody against the N-terminal peptide of cytoplasmic gamma actin. Western blot analysis using SDS-PAGE and isoelectric focusing (IEF) gel revealed that the antibody reacted only with the actin isoforms having gamma motility, confirming that the antibody is specific to the cytoplasmic (nonmuscle) gamma actin. Immunohistochemical analysis of the skeletal muscle of the adult mouse revealed a dot-like staining pattern of the antibody in transverse sections and a striated staining pattern in longitudinal sections. The double immunostaining technique revealed the colocalization of cytoplasmic gamma actin with alpha-actinin, implying the localization of the actin on the Z-disc. Contrary to previous findings (1), we did not detect the colocalization of cytochrome oxidase, a mitochondria marker, with this actin. PMID- 11485323 TI - ApoE(-/-) mice develop atherosclerosis in the absence of complement component C5. AB - Previous studies have suggested that the terminal complex of complement may contribute to the pathogenesis of atherosclerosis. C5b-9 complexes colocalize with the extracellular lipid in the aortic intima of hypercholesterolemic rabbits, and C6-deficient rabbits develop less atherosclerosis than controls. To test the role of complement in atherosclerosis in a different animal model, C5 deficient (C5def) mice were cross-bred with atherosclerosis susceptible apoE(-/-) mice, generating mice deficient in both apoE and C5 and control apoE(-/-) mice. Progeny were typed for C5 titer and serum cholesterol levels. Both male and female mice were fed a high fat diet from weaning until 22 weeks of age. At that time there were no significant differences in plasma cholesterol or triglycerides between apoE(-/-) control and apoE(-/-)/C5def groups. Morphometric analysis of the aortic root lesions gave mean (+/-SEM) lesion areas for male apoE(-/-) and apoE(-/-)/C5def mice of 468,176 +/- 21,982 and 375,182 +/- 53,089 microm(2), respectively (n = 10 each, P value = 0.123). In female apoE(-/-) mice (n = 5), the mean lesion area was 591,981 +/- 53,242 microm(2), compared to 618,578 +/- 83,457 microm(2) for female apoE(-/-)/C5def mice (n = 10) (P value = 0.835). Thus neither male nor female mice showed a significant change in lesion area when C5 was not present. In contrast to the case in the hypercholesterolemic rabbit, activation of the terminal complex of complement does not play a major role in the development of atherosclerosis in apoE(-/-) mice. PMID- 11485324 TI - Assessment of substrate specificity of hepatitis G virus NS3 protease by a genetic method. AB - The RNA genome of hepatitis G virus (HGV) encodes a large polyprotein that is processed to mature proteins by viral-encoded proteases. The HGV NS3 protease is responsible for the cleavage of the HGV polyprotein at four different locations. No conserved sequence motif has been identified for the cleavage sites of the NS3 protease. To determine the substrate specificity of the NS3 protease, amino acid sequences cleaved by the NS3 protease were obtained from randomized sequence libraries by using a screening method referred to as GASP (Genetic Assay for Site specific Proteolysis). Based on statistical analyses of the obtained cleavable sequences, a consensus substrate sequence was deduced: Gln-Glu-Thr-Leu-Val downward arrow Ser, with the scissile bond located between Val and Ser. The relevance of this peptide as a cleavable substrate was further supported by molecular modeling of the NS3 protease. Our result would provide an insight on the molecular activity of the NS3 protease and may be useful for the design of substrate-based inhibitors. PMID- 11485326 TI - Functional abnormalities in protein tyrosine phosphatase epsilon-deficient macrophages. AB - Protein tyrosine phosphatase epsilon (PTP epsilon)-deficient mice were generated by targeted deletion of exons 3, 4, and 5 of the Ptpre gene. Mice homozygous for this deletion (Ptpre(Delta3-5)) were fertile, bred and developed normally and exhibited no overt phenotype. However, closer examination of the function of macrophages from these mice revealed a defect in the regulation of the respiratory burst. While bacterial lipopolysaccharide (LPS) or tumour necrosis factor alpha (TNFalpha) were able to prime bone marrow-derived macrophages (BMM) from wild type (Ptpre(+)) macrophages for an enhanced respiratory burst, they were unable to do so in macrophages from PTP epsilon-deficient mice. PTP epsilon deficient BMM also had abnormalities in cytokine production with a reduced ability to produce TNFalpha and enhanced IL-10 production in response to challenge with LPS. These findings suggest an important role for PTP epsilon in the control of macrophage function. PMID- 11485325 TI - Sterol regulatory element-binding proteins induce an entire pathway of cholesterol synthesis. AB - To evaluate the effects of sterol regulatory element-binding proteins (SREBPs) on the expression of the individual enzymes in the cholesterol synthetic pathway, we examined expression of these genes in the livers from wild-type and transgenic mice overexpressing nuclear SREBP-1a or -2. As estimated by a Northern blot analysis, overexpression of nuclear SREBP-1a or -2 caused marked increases in mRNA levels of the whole battery of cholesterogenic genes. This SREBP activation covers not only rate-limiting enzymes such as HMG CoA synthase and reductase that have been well established as SREBP targets, but also all the enzyme genes in the cholesterol synthetic pathway tested here. The activated genes include mevalonate kinase, mevalonate pyrophosphate decarboxylase, isopentenyl phosphate isomerase, geranylgeranyl pyrophosphate synthase, farnesyl pyrophosphate synthase, squalene synthase, squalene epoxidase, lanosterol synthase, lanosterol demethylase, and 7 dehydro-cholesterol reductase. These results demonstrate that SREBPs activate every step of cholesterol synthetic pathway, contributing to an efficient cholesterol synthesis. PMID- 11485327 TI - Kinetics study of a selenium-containing ScFv catalytic antibody that mimics glutathione peroxidase. AB - The steady-state kinetics study and some enzymatic characterization of a selenium containing scFv catalytic antibody (Se-scFv2F3) were carried out. A novel reaction formula of this abzyme-catalyzed reaction was proposed and a rate equation was obtained according to the formula. The constants in the equation were compared with Dalziel's parameters and the exact meanings of these constants were analyzed. The obtained kinetics parameters from the kinetics study of Se scFv2F3 were analyzed and compared with those of native glutathione peroxidase. PMID- 11485328 TI - A critical role for Syk in endothelial cell proliferation and migration. AB - Syk is a protein-tyrosine kinase that is widely expressed in haematopoietic cells and involved in coupling activated immunoreceptors to downstream signaling. On the other hand, Syk-deficient mice showed severe petechiae in utero and died shortly after birth. Recently we have shown the expression of Syk in endothelial cells and morphological defects of these cells in embryonic Syk-deficient mice. Here we report that both proliferation and migration of human umbilical vein endothelial cells were severely impaired by adenovirus-mediated expression of Syk dominant negative mutants. Furthermore, a close relationship between Syk kinase activity and extracellular signal-regulated kinase activation was suggested. Our results indicate that Syk plays a critical role in endothelial cell functions, including morphogenesis, cell growth, migration, and survival, and contributes to maintaining vascular integrity in vivo. PMID- 11485329 TI - Mice lacking serum amyloid P component do not necessarily develop severe autoimmune disease. AB - Serum amyloid P component (SAP) is a major acute-phase reactant in mice. Recently, it was reported that SAP-deficient mice spontaneously developed anti nuclear antibodies and severe glomerulonephritis. Because the SAP-deficient mice we generated display no obvious phenotypic abnormalities, we investigated whether our SAP-deficient mice would also spontaneously develop autoimmune responses. In accordance with the report, our mice produced high titers of anti-nuclear antibody but did not develop severe glomerulonephritis. On the other hand, it was recently reported that SAP bound to gram-negative bacteria via lipopolysaccharide (LPS) prevented LPS-mediated activation of a classical complement pathway. Thus, we asked if SAP-deficient mice would show altered responses to an intraperitoneal injection of LPS from Salmonella typhimirium. SAP-deficiency did afford resistance to lethality induced by high-dose LPS. Our experiments clearly showed that contrary to documented data, SAP-deficient mice do not develop serious autoimmune disease and we suggest that SAP has a critical role in LPS toxicity. PMID- 11485330 TI - Increased antiviral and opsonic activity of a highly multimerized collectin chimera. AB - Altering the carbohydrate binding properties of surfactant protein D (SP-D) [e.g., by replacing its carbohydrate recognition domain (CRD) with that of either mannose binding lectin (MBL) or conglutinin] can increase its activity against influenza A virus (IAV). The current study demonstrates that the degree of multimerization of SP-D is another independent determinant of antiviral activity. A chimeric collectin containing the N-terminus and collagen domain of human SP-D and the CRD of MBL formed high-molecular-weight multimers similar to those previously described for human SP-D. Using several complementary assays, and diverse viral strains, the chimeric multimers showed greater anti-IAV activity than similarly multimerized preparations of SP-D or incompletely oligomerized preparations of the chimera. More highly multimerized preparations of the chimera also caused greater increases in uptake of IAV by neutrophils. These studies may have implications for development of collectins as therapeutic agents and understanding of natural variations in susceptibility to IAV infection. PMID- 11485331 TI - The full-length and N-terminal deletion of ORF2 protein of hepatitis E virus can dimerize. AB - Hepatitis E virus is a human RNA virus containing three open reading frames. Of these ORF2 encodes, the major capsid protein (pORF2), may possess regulatory functions, in addition to a structural one. In this study, we have shown using the yeast two-hybrid system and in vitro immobilization experiments that full length pORF2 is capable of self-association, thus forming a homodimer. Using mutational analysis we have studied dimerization of various truncated versions of the ORF2 capsid protein using the yeast two-hybrid system and supported our findings with in vitro immobilization experiments. Deletions of pORF2 reveal a loss of the dimerization potential for all deletions except an N-terminal 127 amino-acid deletion. Our studies suggest that the dimerization property of pORF2 may not be amino-acid sequence-dependent but instead a complex formation of a specific tertiary structure that imparts pORF2 its property to self-associate. PMID- 11485332 TI - Novel amidohydrolytic reactions in oxidative pyrimidine metabolism: analysis of the barbiturase reaction and discovery of a novel enzyme, ureidomalonase. AB - Amidohydrolytic reactions in oxidative pyrimidine metabolism were investigated in detail. Barbiturase has been reported to catalyze the amidohydrolysis of barbituric acid to urea and malonic acid. However, purification of the enzyme revealed that it catalyzes the ring-opening of barbituric acid to ureidomalonic acid. The existence of a consecutive enzyme named ureidomalonase, which hydrolyzes ureidomalonic acid to urea and malonic acid, was also discovered during the purification of barbiturase. PMID- 11485333 TI - Epidemiology of osteoporosis. AB - The epidemiology of osteoporosis is reviewed in this article. Attempts were made to answer the following questions: How should osteoporosis be defined? How can risk factors and bone mineral density (BMD) measurements be applied to diagnose osteoporosis? How do the rates for osteoporotic fractures vary by country, sex, age and time? What are the costs for osteoporosis in terms of direct and indirect cost, morbidity and mortality? According to the WHO criteria, osteoporosis can be defined as a BMD of 2.5 standard deviations or more below the young normal mean. BMD measurements are predictive of fracture risks. Hip fracture is by far the most costly of osteoporotic fractures, and the rates are highest in Caucasians, intermediate in Asians and lowest in Blacks. Risk factors could be used to assist in the decision to measure BMD. PMID- 11485334 TI - How to diagnose the presence of osteoporosis and assess the risk of fracture. AB - The approach to the diagnosis of osteoporosis has undergone a radical change in recent years. Osteoporosis is defined as a state of microarchitectural deterioration of the skeleton predisposing that skeleton to fragility fracture. In this sense it is a risk factor for disease in the same way as hypertension is a risk factor. Previously it was considered that bone density measurements using X-ray technology was the best non-invasive method of measuring the microarchitectural deterioration which thus came to define the disease. With the advent of new technology and a more logical approach to risk evaluation this simple approach is no longer appropriate. This chapter takes a stepwise approach to assessing the likelihood of osteoporosis on the basis of clinical factors and then suggests a method of integrating this information with a bone density measure to achieve a prediction of future fracture probability. Methods of diagnosing the cause of the osteoporosis are also outlined. Finally, application of these techniques in clinical care is discussed. PMID- 11485335 TI - Future methods in the assessment of bone mass and structure. AB - There have been major advances in the diagnosis of osteoporosis over the last few decades not only in the definitions that are now used but also in the technology that is available. The future will see further development of the techniques currently in common clinical use, such us dual energy X-ray absorptiometry and quantitative ultrasound. In addition new techniques for assessing bone structure, including MRI and fractal analysis of X-rays, may add significantly to our understanding of the pathophysiology of osteoporosis and to the prediction of fracture risk. PMID- 11485336 TI - Role of biochemical markers in the management of osteoporosis. AB - Several serum and urine biochemical markers of bone resorption and formation have been developed. Biochemical bone markers have been used as intermediate end points in all major studies of anti-osteoporotic therapies. Bone resorption markers, in particular, may add an independent, predictive value to the assessment of bone loss and fracture risk. There are also potential advantages in monitoring anti-osteoporotic treatment in the short-term in addition to bone densitometry, to rapidly identify non-responders to therapy, or non-compliance. Despite these recent advances, until now bone markers have simply been very useful research tools, with their clinical utility being limited by intra individual and diurnal variability. However, the probability of the true bone mineral density response to hormone replacement therapy for the individual patient may be predicted using algorithms based on a spectrum of cut-off bone marker levels with varying false positive and negative rates. Thus, the transition of biochemical bone markers into everyday clinical practice may be rapidly approaching. PMID- 11485337 TI - Corticosteroid osteoporosis. AB - Corticosteroids are widely used and effective agents for the control of many inflammatory diseases, but corticosteroid osteoporosis is a common problem associated with their long term high dose use. Prevention of corticosteroid osteoporosis is preferable to treatment of established corticosteroid bone loss. Several large double-blind controlled clinical trials in patients with corticosteroid osteoporosis have recently been published that provide new insights into its treatment. Based upon available evidence, the rank order of choice for prophylaxis would be a bisphosphonate followed by a vitamin D metabolite or an oestrogen type medication. Calcium alone appears to be unable to prevent rapid bone loss in patients starting corticosteroids, especially with prednisolone doses at 10 mg a day or greater. If an active vitamin D metabolite is used, calcium supplementation should be avoided unless dietary calcium intake is low. Hormone replacement therapy should be considered if hypogonadism is present. Since vertebral fracture is a common and important complication of high dose corticosteroid therapy, these findings suggest that rapid bone loss and hence fractures, can be prevented by prophylactic treatment. Although the follow up data is limited, it is likely that such therapy needs to be continued beyond 12 months whilst patients continue significant doses of corticosteroid therapy. PMID- 11485338 TI - Osteoporosis in men. AB - Osteoporosis is characterized by a reduction in bone density, associated with skeletal fragility and an increased risk of fracture after minimal trauma. Although osteoporosis is generally considered to be a condition affecting post menopausal women, it is now clear that substantial bone loss occurs with advancing age in men, such that up to 20% of symptomatic vertebral fractures and 30% of hip fractures occur in men. This chapter highlights the incidence and prevalence of osteoporotic fractures in men and reviews the associated morbidity, excess mortality and health and social service expenditure. The determinants of peak bone mass and bone loss in men are discussed, as is the pathogenesis of osteoporosis and vertebral and hip fractures. The criteria for the diagnosis of osteoporosis in men are reviewed, together with the most appropriate investigations for secondary osteoporosis. The management of osteoporosis in men is also discussed, highlighting the most appropriate treatment options. PMID- 11485339 TI - The evidence that exercise during growth or adulthood reduces the risk of fragility fractures is weak. AB - There has never been, and will never be, a randomized double-blind placebo controlled trial demonstrating that exercise in youth, adulthood or old age reduces fragility or osteoporosis-related fractures in old age. The next level of evidence, a randomized, controlled but unblinded study with fractures as an end point is feasible but has never been done. The basis for the belief that exercise reduces fractures is derived from lower levels of 'evidence', namely, retrospective and prospective observation cohort studies and case-control studies. These studies are at best hypothesis generating, never hypothesis testing. They are all subject to many systematic biases and should be interpreted with extreme scepticism. Surrogate measures of anti-fracture efficacy are the next level of evidence, such as the demonstration of a reduction in risk factors for falls, a reduction in falls, a reduction in fractures due to falls, an increase in peak bone size and mass, prevention of bone loss in midlife and restoration of bone mass and structure in old age. PMID- 11485340 TI - Oestrogen and selective oestrogen receptor modulators (SERMs): current roles in the prevention and treatment of osteoporosis. AB - Post-menopausal oestrogen replacement therapy (HRT) is the agent of choice for restraint of post-menopausal bone loss. In numerous observational, and in a few intervention studies, it has been shown to reduce osteoporotic fractures. However, in order to be effective treatment has to be carried on long term. This is a task not easily achieved due to poor patient adherence. The majority of patients stop treatment after a few months. Fear of breast cancer and refusal to tolerate resumption of cyclic withdrawal bleeding are the principal objections. Newer and more user-friendly forms of treatment are needed. Ideally, these should have an efficacy similar to oestrogen on the central nervous system, cardiovascular system and bone, without adverse effects on the breast and uterus. With this in mind, Selective oestrogen Receptor Modulators (SERMs) have been developed. In this chapter we review how effective modern pharmacology has been in achieving an improved design of hormonal replacement for the post-menopausal women, and the place that it has obtained today alongside both older and newer forms of treatment. PMID- 11485341 TI - Role of bisphosphonates and calcitonin in the prevention and treatment of osteoporosis. AB - Bisphosphonates have been shown to increase bone mineral density in patients with established osteoporosis as well as those with osteopenia. The evidence conclusively shows a reduction in fracture rates in patients on the more potent nitrogen containing bisphosphonates. Indeed, significant vertebral fracture rate reduction has been demonstrated after only 1 year of therapy. Alendronate, a second-generation bisphosphonate, and risedronate, a third-generation bisphosphonate, are first line medications for the treatment of osteoporosis given their efficacy in preventing both vertebral and non-vertebral fractures. There is evidence that vertebral fractures may be prevented by intermittent cyclic therapy with etidronate. All three have been shown to increase bone mineral density in the spine, with alendronate and risedronate producing significant increases in hip bone density. Calcitonin has demonstrated the ability to reduce vertebral fracture rates with minimal changes in bone density. Calcitonin is also beneficial in reducing the bone pain associated with fractures. PMID- 11485342 TI - Emerging therapies in osteoporosis. AB - The approval of alendronate in 1994 marked a watershed in the treatment of osteoporosis. Before that time there was no therapy for which unequivocal proof of efficacy existed. Since then several more agents, all from the anti-resorptive class, have also been approved for use in the treatment of this disease and the range of indications for alendronate has been extended to include the prevention of osteoporosis in women with lesser degrees of bone loss, the treatment of glucocorticoid-induced osteoporosis and, most recently, the treatment of male osteoporosis. Despite this there are still several areas of unmet medical need in this disease, including the availability of well tolerated and convenient therapies and treatments that will go beyond the levels of efficacy offered by current therapies. An intense effort is now being directed towards meeting these unmet needs with the improvement of existing therapies and the development of novel agents that will provide superior long term benefit. Important and exciting drug targets are yielding novel compounds with anti-resorptive activity or anabolic effects to complement current anti-resorptives. Despite this effort considerable obstacles to the successful development of these compounds remain, not least the stringent safety requirements needed to provide an acceptable risk to-benefit profile and the increasing difficulties of conducting placebo controlled studies in patients at high risk of fracture. PMID- 11485343 TI - How to prevent fractures in the individual with osteoporosis. AB - The high rate of osteoporotic fracture in Western populations has resulted in a significant burden in terms of morbidity, mortality and health care costs. The use of DXA has made the diagnosis of osteoporosis easier and identified a subgroup of individuals who are at a higher risk of fracture. It is a useful tool in determining therapy in those at greatest risk of fracture. However, widespread use of such treatments is low and greater uptake remains an elusive goal. There are now many different treatments that reduce fracture rate, and can accompany lifestyle measures such as smoking cessation, diet and exercise. Dietary supplementation with calcium has been shown to reduce the risk of vertebral fracture, and the combination of calcium with vitamin D has been shown to reduce fracture at non-vertebral sites, including the hip. Although ERT, SERMs and tibolone all retard bone loss, prospective fracture prevention has only been shown for SERMs and then only at the spine. Bisphosphonates represent a class of potent anti-resorptive agents, which have been shown to reduce fracture rate at vertebral and non-vertebral sites. Other agents such as calcitonin, PTH and fluoride are of less certain benefit in preventing fracture. PMID- 11485346 TI - Severe impairment of primary but not memory responses to influenza viral antigens in aged mice: costimulation in vivo partially reverses impaired primary immune responses. AB - Profound alterations in humoral and cellular immune responses are a hallmark of aging, and understanding the immunobiology of aging is key to the success of preventive vaccination strategies. With aging, while recall or memory responses to influenza viral antigens for the most part remained unaltered, primary immune responses are severely impaired. The impaired primary responses are partly due to a lack of costimulation, as providing costimulation at the time of induction of primary immune responses against influenza virus vaccine partially reversed aged related immune dysfunction and conferred enhanced protection. Inclusion of immunomodulators that up-regulate the expression of costimulatory molecules must be considered to improve the efficacy of vaccination in the elderly, particularly to novel immunogens. PMID- 11485347 TI - Bromelain activates murine macrophages and natural killer cells in vitro. AB - The innate immune response is critical for effective immunity against most pathogens. In this study, we show that bromelain, a mixture of cysteine proteases, can enhance IFN-gamma-mediated nitric oxide and TNFalpha production by macrophages. Bromelain's effect was independent of endotoxin receptor activation and was not caused by direct modulation of IFN-gamma receptors. Instead, bromelain either enhanced or acted synergistically with IFN-gamma receptor mediated signals. These effects were seen in both RAW 264.7, a macrophage cell line, and primary macrophage populations. Bromelain also increased IL-2- and IL 12-mediated IFN-gamma production by NK cells. These results indicate a potential role for bromelain in the activation of inflammatory responses in situations where they may be deficient, such as may occur in immunocompromised individuals. PMID- 11485348 TI - Monocyte differentiation and HIV replication after prolonged culture of peripheral blood mononuclear cells from HIV-infected individuals. AB - Primary cultures of peripheral blood mononuclear cells (PBMC) from 51 HIV+ hemophiliac patients (HIV+ PBMC) were set up, allowing undisturbed cellular interaction in the absence of any exogenous stimuli. The optimum time for p24 detection was between 12 and 25 days. Infective virus was recovered from the culture supernatants (HIV+ SN) and the amount of p24 released ranged from 25 to 5300 pg/ml. Cells of the monocyte/macrophage (M/M) lineage were the main source of HIV in the HIV+ SN, as judged by intracellular staining of permeabilized cells with anti-p24 (KC57 monoclonal antibody) and flow cytometry analysis. M/M activation, differentiation, and proliferation occurred along the culture before the peak of in vitro HIV replication. Release of HIV p24 was highest in patients with >200 CD4+ T lymphocytes/mm3 who did not receive highly active antiretroviral therapy (HAART), but it was still detectable in 60-90% of patients who had responded to 1-2 years of HAART, reducing their plasma viral load to undetectable levels. It is proposed that this simple experimental system can be used to assess ongoing HIV infection of M/M with the patient's own viral variants. PMID- 11485349 TI - The granzyme B inhibitor, PI-9, is present in endothelial and mesothelial cells, suggesting that it protects bystander cells during immune responses. AB - Proteinase inhibitor 9 (PI-9) is a 42-kDa human intracellular serpin present in cytotoxic lymphocytes (CLs). PI-9 is an extremely efficient inhibitor of the pro apoptotic CL granule proteinase granzyme B and is thought to function in the cytosol of CLs to protect against apoptosis induced by endogenously expressed or released granzyme B, particularly during target cell killing. Here we show by immunohistochemistry that PI-9 is also present in endothelial cells, in every tissue examined. Cultured endothelial cells express functional PI-9 (as assessed by binding to recombinant granzyme B) localized to the cytoplasm and nucleus. Immunohistochemistry also showed PI-9 in mesothelial cells, and this was confirmed by analysis of primary cells cultured from pleural and serous effusions. Granzyme B expression was not detected in either endothelial or mesothelial cells. In both cell types, PI-9 is up-regulated at the mRNA and protein level by exposure to the phorbol ester PMA, consistent with a response to inflammatory stimuli. We postulate that PI-9 is present in these lining cell types to protect against misdirected, free granzyme B released during a local immune response. PMID- 11485350 TI - Costimulation by extracellular matrix proteins determines the response to TCR ligation. AB - Although ligation of the T-cell antigen receptor (TCR) is central to the responsiveness and antigen specificity of T-cells, it is insufficient to elicit a response. To determine whether the need for costimulation reflects inadequate strength of signal transduction through the TCR or an absolute block of signaling in the absence of a coligand, we studied T-cell activation under serum-free conditions eliminating costimulation by various extracellular matrix proteins which otherwise have an omnipresent and frequently overlooked effect. Engagement of the TCR leads to induction of Fas, but not to measurable IL-2 secretion or apoptosis. Those activation parameters are induced by costimulation through integrin alphaVbeta3. Furthermore, T-cell survival or elimination is determined by the type of ligand binding to this coreceptor with vitronectin, fibronectin, and fibrinogen efficiently inducing apoptosis and IL-2 production while osteopontin and entactin mediate IL-2 secretion comparably without causing programmed cell death. Consistent with the cytokine properties of these ligands, differential costimulation depends on their presentation in soluble rather than immobilized form. The determination of elimination versus survival of activated T cells by coligation of beta3-integrins may have bearing on the fundamental postthymic mechanisms that shape the T-cell repertoire. PMID- 11485351 TI - Rat hepatic natural killer cells (pit cells) express mRNA and protein similar to in vitro interleukin-2 activated spleen natural killer cells. AB - Pit cells are liver-specific natural killer (NK) cells that can be divided into high- (HD) and low-density (LD) subpopulations. The characteristics of pit cells were further investigated in this report. LD and HD pit cells express the specific NK-activation markers gp42, CD25, and ANK44 antigen. LD cells and IL-2 activated NK cells have a high mRNA expression of perforin, granzymes, interferon gamma, and tumor necrosis factor-alpha. LD pit cells, unlike spleen NK cells, have a weak response to IL-2 with regard to proliferation, cytotoxicity, and production of NK-related molecules. The characteristics of HD cells are intermediate between LD and spleen NK cells. These results show that pit cells, especially LD cells, possess characteristics similar to IL-2-activated NK cells. This is the first evidence on a molecular level that pit cells could be considered in vivo activated NK cells. PMID- 11485352 TI - Liposome mediated antigen delivery leads to induction of CD8+ T lymphocyte and antibody responses against the V3 loop region of HIV gp120. AB - There is general consensus that the use of whole viruses for the development of a vaccine against human immunodeficiency virus (HIV) would be unsafe. While currently available nonreplicating vaccines, composed of synthetic peptides or purified subunit antigens, can help in tricking the humoral immune responses, they fail to incite the other major arm of the immune defense system, i.e., cell mediated immunity (CMI). To overcome the difficulty in generating CMI, we have entrapped an immunodominant HIV envelope glycoprotein peptide in liposomes made up of fusogenic lipids isolated from Escherichia coli. We have established the role of fusogenic liposomes in stimulation of HIV-specific CD8+ cytotoxic T lymphocytes. Interestingly, the same liposomes elicit strong HIV-specific antibody production as well. Moreover, untoward manifestations such as skin damage or antibody production against lipid components were also not observed. Thus, E. coli lipid liposomes (escheriosomes) could prove to be a potent candidate vaccine, capable of eliciting both humoral and cell mediated immune responses against HIV infection. PMID- 11485353 TI - 4-1BBL enhances anti-tumor responses in the presence or absence of CD28 but CD28 is required for protective immunity against parental tumors. AB - A20 is an aggressive BALB/c B cell lymphoma that, despite its expression of B7-2, rapidly forms tumors in syngeneic mice. We have generated A20 transfectants expressing elevated levels of B7-2 (A20/B7-2high) or 4-1BBL (A20/4 1BBL(low,mod,high)) and found that mice which were able to reject the A20/B7-2 or A20/4-1BBL transfectants were also resistant to subsequent systemic challenge with the parental cell line. To assess whether the effectiveness of 4-1BBL in enhancing anti-tumor immunogenicity was dependent on additional signals from B7 CD28 interaction, we injected the A20 variants into BALB/c CD28(-/-) mice. We found that CD28(-/-) mice were able to reject the A20/4-1BBL variants while A20/B7-2 cells formed tumors. However, when the A20/4-1BBL resistant CD28(-/-) mice were systemically challenged with the A20 parental line, tumors formed rapidly. Upon restimulation in vitro, splenocytes from A20/4-1BBL immunized CD28(+/+) mice were able to kill parental tumors whereas splenocytes from CD28(-/ ) mice showed a reduction in CTL activity against A20 or A20/4-1BBL targets. Examination of cytokine production by the immunized animals indicated that the CD28(-/-) splenocytes secreted substantially less IL-2 as well as reduced levels of IFN-gamma compared with their CD28(+/+) counterparts. Thus, 4-1BBL expressing tumors are capable of priming CTL responses against 4-1BBL transfected as well as parental tumors in the absence of CD28. However, in the absence of CD28 signaling, the production of cytokines and particularly IL-2 was lower, resulting in a weaker CTL recall response and reduced ability to survive challenge with parental tumor. PMID- 11485354 TI - Bromelain modulates T cell and B cell immune responses in vitro and in vivo. AB - The ability to modulate immune responses is a major aim of many vaccine and immunotherapeutic development programs. Bromelain, a mixture of cysteine proteases, modulates immunological responses and has been proposed to be of clinical use. However, the identity of the immune cells affected by bromelain and the specific cellular functions that are altered remain poorly understood. To address these shortcomings in our knowledge, we have used both in vitro and in vivo immunological assays to study the effects of bromelain. We found that bromelain enhanced T cell receptor (TCR) and anti-CD28-mediated T cell proliferation in splenocyte cultures by increasing the costimulatory activity of accessory cell populations. However, despite increased T cell proliferation, bromelain concomitantly decreased IL-2 production in splenocyte cultures. Additionally, bromelain did not affect TCR and CD28-induced proliferation of highly purified CD4+ T cells, but did inhibit IL-2 production by these cells. In vivo, bromelain enhanced T-cell-dependent, Ag-specific, B cell antibody responses. Again, bromelain induced a concomitant decrease in splenic IL-2 mRNA accumulation in immunized mice. Together, these data show that bromelain can simultaneously enhance and inhibit T cell responses in vitro and in vivo via a stimulatory action on accessory cells and a direct inhibitory action on T cells. This work provides important insights into the immunomodulatory activity of bromelain and has important implications for the use of exogenous cysteine proteases as vaccine adjuvants or immunomodulatory agents. PMID- 11485355 TI - Static biomechanical evaluation of the foot and lower limb: the podiatrist's perspective. AB - SUMMARY. A biomechanical assessment is one aspect of podiatry. It involves two main modes of examination: dynamic and static. Aspects of the static mode are described with particular emphasis on the foot. The static mode includes both open and closed kinetic chain examinations. Each part of the lower extremity is evaluated with reference to the planes of the body. In open chain examination specific techniques have been developed to evaluate the quality and range of motion of the foot joints. The foot is held in a standardised position which simulates the midstance period of the gait cycle. The position and function of the subtalar joint is considered particularly influential in the foot. For this reason the method of manipulating the patient's subtalar joint into the 'neutral position' is important. In closed chain examination, posture is evaluated in both 'relaxed' and 'neural' stance positions. Dynamic and static biomechanical examination data has to be interpreted with reference to the primary complaint and a full medical history, together with specific information concerning footwear and the habitual locomotor functional needs of the patient. Copyright 1997 Harcourt Publishers Ltd. PMID- 11485356 TI - Outcomes assessment in musculoskeletal medicine. AB - SUMMARY. Outcomes management is important for quality assurance purposes. Issues of reliability and validity are central to determining appropriate outcomes tools. Outcomes can be used to measure progress in three critical areas of patient management: pain relief, physical capacity (impairment), and disability. Outcomes can be obtained inexpensively and in a time-efficient manner. This article reviews the most valuable outcomes tools for a small, private practice and how they can be implemented into patient care. Copyright 1997 Harcourt Publishers Ltd. PMID- 11485357 TI - Instigators of activity intolerance. AB - SUMMARY. During the course of this century several Western societies have experienced an insufficiently explained exponential increase in disability due to back pain. Recently the International Association for the Study of Pain's Task Force on Back Pain in the Work Place summed up the present problem and made recommendations for its future management. Management, and possibly prevention, are likely to be assisted by understanding the reasons responsible for this unsustainably costly phenomenon. It is proposed that one such reason involves maladaptive fear-avoidance beliefs and deconditioning behaviour arising out of convictions as to some structural-anatomical-biomechanical (SAB) basis for the pain. Also, these beliefs and convictions may receive neurophysiological endorsement from peripheral afferent input that is normally not registered as being 'pathologically'/clinically painful.The Task Force's proposed 'demedicalization' of non-specific low back pain (NSLBP) together with the endorsement of (chiropractic) 'manipulation' will probably help reduce the immediate costs of invasive treatments and/or hospitalization as well as the deconditioning consequences of prolonged inactivity. However, it is contended that, in the longer term, it would also be necessary to deal effectively with maladaptive fear-avoidance beliefs and SAB-based convictions as to the cause, meaning and management of back pain. In some Western societies (manipulative) physiotherapists may be given the opportunity to be a part of this 'demedicalization' process. Therefore, it would now be appropriate to consider the issues involved, some of which are raised here, in order to help decide what direction the physiotherapy profession might take. Copyright 1997 Harcourt Publishers Ltd. PMID- 11485358 TI - Inter-therapist reliability in locating latent myofascial trigger points using palpation. AB - SUMMARY. Myofascial trigger points (TPs) are sites in muscle or fascia which are tender to palpate and are located in a taut band of tissue. The significance of TPs is underlined by claims that treatment applied to TPs can be effective in treating pain. Before such claims can be validated, the ability of clinicians to identify these points reliably using palpation needs to be established. The location of such sites is also important for studies which try to determine the relationship of these points to other measures of pathology, e.g. EMG and thermographic abnormalities. Intertester reliability in the localization of these points, therefore, appears to be of primary importance. This study examined the concordance between two experienced clinicians in being able to identify TPs in the upper trapezius muscle. The results indicated that the agreement between these clinicians in identifying TPs in asymptomatic subjects was poor. There was a lack of concordance not only in the location of sites of TPs but also in the numbers of TPs identified. This outcome challenges claims that TPs can be reliably identified using palpation. Copyright 1997 Harcourt Publishers Ltd. PMID- 11485359 TI - Upper cervical instability: are clinical tests reliable? AB - SUMMARY. The aim of this preliminary study was to investigate the reliability of a selection of manual tests used for the examination of instability of the upper cervical region. Eleven children with Down's syndrome were examined by four independent examiners with different levels of experience in manual therapy. Three tests as described by Van der El (1992) were used: the lateral displacement test, the Sharp-Purser test, and the upper cervical flexion test. Scores of tests and retests were statistically analysed by calculating the percentage of agreement, and Brennan and Prediger's modified kappa, and with the binomial test. The results showed a significant agreement between test and retest for the upper cervical flexion test in three out of four investigators. Agreement between investigators was significant in four out of six combinations between two investigators, and near to significant for this test (P(bin) =.07) in the two remaining combinations. The other two investigated tests and the total score showed no tendancy towards a consistent level of significant intra- or interobserver reliability. Copyright 1997 Harcourt Publishers Ltd. PMID- 11485360 TI - Relationship between the cervical component of the slump test and change in hamstring muscle tension. AB - SUMMARY. The slump test has been used routinely to differentiate low back pain due to involvement of neural structures from low back pain attributable to other factors. It is also said to differentiate between posterior thigh pain due to neural involvement from that due to hamstring injury. If changes in cervical position affect the hamstring muscles, differential diagnosis is confounded. Posterior thigh pain caused by the cervical component of the slump could then be caused either by increased tension on neural structures or increased tension in the hamstrings themselves. The aim of this study was to determine whether changing the cervical position during slump altered posterior thigh pain and/or the tension in the hamstring muscle. Asymptomatic subjects aged between 18 and 30 years were tested. A special fixation device was engineered to fix the trunk, pelvis and lower limb. Pain levels in cervical flexion and extension were assessed by visual analogue scale. Fixation was successful in that there were no significant differences in position of the pelvis or knee during changes in cervical position. Averaged over the group, there was a 40% decrease (P < 0.05) in posterior thigh pain with cervical extension. There were no significant differences in hamstring electromyographic readings during the cervical movements. This indicated that: (1) cervical movement did not change hamstring muscle tension, and (2) the change in experimentally induced pain during cervical flexion was not due to changes in the hamstring muscle. This conclusion supports the view that posterior thigh pain caused by the slump test and relieved by cervical extension arises from neural structures rather than the hamstring muscle. Copyright 1997 Harcourt Publishers Ltd. PMID- 11485367 TI - Nitric oxide inhibits the cochaperone activity of the RING finger-like protein DnaJ. AB - As a consequence of bacterial infection and the ensuing inflammation, expression of the inducible NO synthase results in prolonged synthesis of NO in high concentrations, which among other functions, contributes to the innate defense against the infectious agent. Here we show that NO inhibits the ability of the bacterial cochaperone DnaJ containing a RING finger-like domain to cooperate with the Hsp70 chaperone DnaK in mediating correct folding of denatured rhodanese. This inhibition is accompanied by S-nitrosation of DnaJ as well as by Zn2+ release from the protein. In contrast, NO has no effect on the activity of GroEL, a bacterial chaperone without zinc sulfur clusters. Escherichia coli cells lacking the chaperone trigger factor and thus relying on the DnaJ/DnaK system are more susceptible toward NO-mediated cytostasis than are wild-type bacteria. Our studies identify the cochaperone DnaJ as a molecular target for NO. Thus, an encounter of bacterial cells with NO can impair the protein folding activity of the bacterial chaperone system, thereby increasing bacterial susceptibility toward the defensive attack by the host. PMID- 11485368 TI - The role of nitric oxide in mediating nonadrenergic, noncholinergic relaxation in rat pulmonary artery. AB - Experiments were undertaken to investigate the existence of inhibitory nonadrenergic, noncholinergic (i-NANC) nerve activity by using in vitro functional and immunohistochemical techniques in rat main pulmonary arterial rings. Vessels precontracted with phenylephrine (3 microM) relaxed in response to electrical field stimulation (EFS) (50 V, 0.2 ms, 0.1-10 Hz for 5 s) in the presence of atropine (1 microM) and guanethidine (1 microM). Tetrodotoxin (0.3 microM) abolished this response, indicating that it is neuronal in origin. l-NAME (30 microM), methylene blue (10 microM), and removal of endothelium significantly reduced the EFS-induced relaxations. The inhibitory action of l-NAME was completely reversed by l-arginine (1 mM) but not by d-arginine (1 mM). Moreover l arginine alone potentiated the magnitude of the relaxations elicited by EFS. On the other hand, immunohistochemical work clearly demonstrated the existence of neuronal nitric oxide synthase in the pulmonary artery vessel wall. All these results are consistent with the suggestion that nitric oxide is the likely mediator of this vasodilatation. However, the incomplete blockade of the responses by l-NAME gives evidence of an additional inhibitory NANC neurotransmitter(s) mediating the residual relaxation, which requires further experiments to clarify its nature. PMID- 11485369 TI - Histochemical and immunohistochemical localization of neuronal nitric oxide synthase in the olfactory epithelium of the axolotl, Ambystoma mexicanum. AB - The aim of this study was to describe the anatomic distribution of neuronal nitric oxide synthase immunoreactivity (nNOS-IR) and nicotinamide-adenine dinucleotide phosphate-diaphorase (NADPH-d) staining in the olfactory epithelium of the axolotl, juvenile, and neotenic adult, Ambystoma mexicanum. Nitric oxide (NO, nitrogen monoxide) is a widespread molecule that has been identified both as a neuromodulator and as an intracellular messenger. In the olfactory system, NO has been proposed to play a role in olfactory transduction. Nitric oxide synthase (NOS) can be detected by histochemical (NADPH-d) and immunohistochemical techniques. NADPH-d staining has been described in olfactory receptor neurons (ORN) of several species; however, nNOS-IR has not always been found at ORN. Present results show intense NADPH-d staining and nNOS-IR in the dendrites and cell bodies of ORN in both the nasal cavity and the vomeronasal organ of axolotls. Unilateral olfactory axotomy was conducted to confirm that labels were at ORN. Two weeks after this procedure an important decrease in NADPH-d staining and nNOS-IR was observed. The remaining labels were mostly in basal cells. By 5 weeks postaxotomy both labels were almost totally absent. Thus, both NADPH-d staining and nNOS-IR were mainly localized in ORN. NADPH-d staining and nNOS-IR were also found in nerve fibers surrounding arterioles, as well as in secretory and duct cells of the Bowman's glands. This last anatomical localization suggests that in the A. mexicanum NO might be involved in functions other than only olfactory transduction, such as regulation of local blood flow, glandular secretion, and ORN development. PMID- 11485370 TI - Ischemia increases detectable endothelial nitric oxide synthase in rat and human myocardium. AB - The aims of the present study were to establish if myocardial ischemia/reperfusion is associated with altered eNOS activity and if myocardial eNOS detection depends on its activity. We determined detectable eNOS in (1) myocardium of isolated perfused rat hearts subjected to either global or regional ischemia and (2) in left ventricular biopsies from patients undergoing two different methods of myocardial protection (i.e., intermittent cold blood cardioplegia and continuous coronary perfusion with warm, beta-blocker-enriched blood) during coronary artery surgery. NOS detection was performed by NADPH-d staining and three eNOS-antibodies against different eNOS epitopes. In addition, activity dependent alteration of detectable eNOS was proofed by bradykinin treatment for 2 to 10 min. Ischemic and receptor mediated eNOS activation increased NADPH-d reactivity and eNOS immunoreaction as measured by antibodies against either amino acids of a central bovine eNOS domain or the human eNOS N terminal end. In contrast, the antibody against the human eNOS C-terminal end exhibited no alteration of eNOS immunoreaction. The transient eNOS activation was associated with increased cGMP content. In human myocardium subjected to ischemia during cardiac surgery we found that early reperfusion increases eNOS activity. These data demonstrate a strong association between myocardial ischemia/reperfusion and increased eNOS activity as measured by immunocytochemical staining against specific eNOS epitopes. It appears that eNOS activation and subsequent NO release may act as a regulatory system to counter balance the potentially deleterious effects of myocardial ischemia/reperfusion. PMID- 11485371 TI - Effects of nitric oxide synthase inhibitors on vascular hyperpermeability with thermal injury in mice. AB - The role of nitric oxide and related synthase in thermal injury was investigated by using models of experimental burn to evaluate severity from the aspect of vascular permeability. Thermal injuries were produced in the murine right ear by pinching with a pair of preheated tweezers. Immediately thereafter, Evans blue dye was intravenously administered, and the mice injured with burns were sacrificed at various times. The burned ears were collected and hydrolyzed, and the level of extracted dye was measured as an indicator of inflammation. Vascular hyperpermeability was suppressed by the administration of nitric oxide synthase inhibitors. LNAME not only suppressed vascular hyperpermeability in thermal injuries in a dose-dependent manner but was also effective with either prophylactic or therapeutic administration. Although aminoguanidine also suppressed the inflammatory response, it had no effect on the early inflammatory phase. Nitric oxide synthase is well known to have two types of isozymes. Aminoguanidine, an inhibitor specific to inducible nitric oxide synthase, suppressed the late phase 6 h after injury, suggesting that inducible nitric oxide synthase is involved in inflammatory responses of thermal injuries. These results also demonstrated that inducible nitric oxide synthase-like protein stained the burned region immunohistochemically. Therefore, both types of enzymes mediating nitric oxide affect inflammatory responses, i.e., vascular hyperpermeability, and their regulation may lead to the development of new therapy for thermal injuries. PMID- 11485372 TI - Association between the NOS3 (-786 T/C) and the ACE (I/D) DNA genotypes and early coronary artery disease. AB - DNA polymorphisms at the endothelium constitutive nitric oxide synthase gene (NOS3) have been linked to the risk of developing coronary artery disease (CAD). In vitro, a polymorphism in the 5' region of the NOS3 gene (-786 T/C) influences promoter activity. This polymorphism has been associated with coronary spasms among Japanese. The genetic variation at the angiotensin-converting enzyme (ACE) is associated with plasma ACE activities and has also been linked with susceptibility to cardiovascular disease. Our objective was to determine if DNA polymorphisms in the NOS3 and ACE genes were associated with early CAD. We analyzed the -786 T/C polymorphism in the 5' flanking region and the 27-bp repeat polymorphism in NOS3 intron 4, as well as the ACE-I/D polymorphism. A total of 170 male smokers (CAD patients) younger than 50 years and 300 male smokers (healthy controls) were genotyped. Frequencies were compared by the chi(2) test, and odds ratios (ORs) and their 95% confidence intervals (CI) were also calculated. Only the -786 T/C polymorphism in the 5' flanking region of the NOS3 gene was significantly associated with early CAD in our population. The frequency of the CC genotype was significantly increased (P = 0.039) in patients compared to controls (OR = 1.67; 95% CI = 1.01, 2.72). We found a synergistic effect between the NOS3-CC and the ACE-DD genotypes in the risk of developing early CAD. The frequency of CC + DD was significantly increased among patients (P = 0.002). Thus, those with a NOS3-CC and an ACE-DD genotype would have a significantly increased risk of suffering an early episode of coronary artery disease (OR = 2.82; 95% CI = 1.40, 5.70). Although based on a limited number of patients, our work suggests that individuals who are NOS3-CC + ACE-DD are at a higher risk for early CAD, probably as a consequence of increased endothelial dysfunction. PMID- 11485373 TI - N-acetylcysteine inhibits in vivo nitric oxide production by inducible nitric oxide synthase. AB - This in vivo study evaluates the effect of N-acetylcysteine (NAC) administration on nitric oxide (NO) production by the inducible form of nitric oxide synthase (iNOS). NO production was induced in the rat by the ip administration of 2 mg/100 g lipopolysaccharide (LPS). This treatment caused: (1) a decrease in body temperature within 90 min, followed by a slow return to normal levels; (2) an increase in plasma levels of urea, nitrite/nitrate, and citrulline; (3) the appearance in blood of nitrosyl-hemoglobin (NO-Hb) and in liver of dinitrosyl iron-dithiolate complexes (DNIC); and (4) increased expression of iNOS mRNA in peripheral blood mononuclear cells (PBMC). Rat treatment with 15 mg/100 g NAC ip, 30 min before LPS, resulted in a significant decrease in blood NO-Hb levels, plasma nitrite/nitrate and citrulline concentrations, and liver DNIC complexes. PBMC also showed a decreased expression of iNOS mRNA. NAC pretreatment did not modify the increased levels of plasma urea or the hypothermic effect induced by the endotoxin. The administration of NAC following LPS intoxication (15 min prior to sacrifice) did not affect NO-Hb levels. These results demonstrate that NAC administration can modulate the massive NO production induced by LPS. This can be attributed mostly to the inhibitory effect of NAC on one of the events leading to iNOS protein expression. This hypothesis is also supported by the lack of effect of late NAC administration. PMID- 11485374 TI - Nitric oxide activates voltage-dependent potassium currents of crustacean skeletal muscle. AB - Nitric oxide (NO), a radical gas, acts as a multifunctional intra- and intercellular messenger. In the present study we investigated the effects of NO on muscle membrane potassium currents of isolated single muscle fibers from the marine isopods, Idotea baltica, using two-electrode voltage clamp recording techniques. Voltage-activated potassium currents consist of an outward current with fast activation and inactivation kinetics and a delayed, persistent outward current. Both currents were blocked by extracellular 4-aminopyridine and tetraethylammonium; the currents were not blocked by charybdotoxin or apamin. Application of the NO donors S-nitroso-N-acetylpenicillamine (SNAP) or hydroxylamine increased both the early and the delayed outward current in a dose- and time-dependent manner. PTIO, a NO scavenger, suppressed the effect of SNAP. N Acetyl-dl-penicillamine, a related control compound which does not liberate NO, had no significant effect on outward currents. Methylene blue, a guanylyl cyclase inhibitor, prevented the increase of the outward current while 8-bromo-cGMP increased the current. Our experiments show that potassium currents of Idotea muscle are increased by NO donors. They suggest that NO by stimulating cGMP production mediates the effects on membrane currents involved in regulation of invertebrate muscle excitability. PMID- 11485375 TI - The renal and systemic hemodynamic effects of a nitric oxide-synthase inhibitor are reversed by a selective endothelin(a) receptor antagonist in men. AB - There is evidence for an interaction between nitric oxide (NO) and endothelin (ET) at the level of the renal vasculature. We hypothesized that acute renal effects of systemic NO synthase inhibition (NG-monomethyl-l-arginine, L-NMMA) may be blunted by coadministration of a specific ET(A) receptor antagonist (BQ-123) in healthy humans. Fifteen healthy young male subjects participated in this randomized, double-blind, placebo-controlled 3-way crossover study. These sodium repleted volunteers received L-NMMA alone, or BQ-123 alone, or L-NMMA with a subsequent coinfusion of BQ-123. Renal plasma flow (RPF) and glomerular filtration rate (GFR) were determined with the PAH and inulin clearance method, respectively. Mean arterial pressure (MAP) and pulse rate were measured noninvasively at baseline and every 15 min after the start of the study period. L NMMA alone reduced RPF (-22%, P < 0.001) and GFR (-8%, P < 0.009) and increased MAP (+10%, P < 0.001). BQ-123 alone did not affect these parameters. However, coinfusion of BQ-123 blunted the effects of L-NMMA on RPF (P < 0.001), GFR (P < 0.001), and MAP (P = 0.006). Peripheral and renal hemodynamic effects of acute systemic NO synthase inhibition are at least partially reversed by ET(A) receptor blockade with BQ-123. This indicates a functional antagonism between specific ET(A) receptor antagonist and NO synthase inhibitors at the level of the renal vasculature. PMID- 11485376 TI - Chemistry of the diazeniumdiolates. I. Structural and spectral characteristics of the [N(O)NO]- functional group. AB - Ions of structure X[N(O)NO]-, examples of which have seen increasing use as probes for studying the biology of nitric oxide (NO) over the past decade, have a varied chemical history spanning nearly two centuries. Nevertheless, they have not been widely appreciated for their physicochemical similarities. Here we begin a series of systematic inquiries into the fundamental chemistry of such compounds aimed at identifying both the characteristics that justify considering them as a group and the factors that contribute to observed differences in their physicochemical properties. In the present paper, X-ray structures in which X is SO3- (1), O- (2), Ph (3), and Et2N (5), as well as that of the gem-disubstituted carbon derivative CH2[N(O)NO]2-(2) (4), are compared. All their O-N-N-O systems are essentially planar, with cis oxygens and an N-N linkage exhibiting considerable double-bond character. The ultraviolet spectrum of the isolated chromophore consists of a relatively intense ( approximately 6-10 mM(-1) x cm(-1) per [N(O)NO]- group) absorption at 248-250 nm (for 2 and 5) that is red shifted by through-space Stark interactions (e.g., by approximately 10 nm in 1 and 4) as well as by conjugative interaction with X (lambda(max) = 284 nm for 3). Infrared and Raman spectra for the widely used pharmacological probe 5 were determined, with analysis of vibrational modes being aided by comparison with the spectra of the [15N(O)15NO]- isotopomer and density functional theory calculations at the B3LYP/6-311++G** level. To address confusion that has arisen in the literature resulting from rather widespread use of differing trivial designations for this class of compounds, a unifying nomenclature system is recommended in which compounds containing the [N(O)NO]- moiety are named as diazeniumdiolates. It is hoped that these and other efforts to understand and predict the physicochemical similarities and differences among different members of the diazeniumdiolate class will aid in reaping their full potential in the area of rational drug design. PMID- 11485377 TI - Reaction mechanism between nitric oxide and glutathione mediated by Fe(III) myoglobin. AB - Ferrimyoglobin at pH 7.4 binds nitric oxide to yield nitric oxide adducts. In the presence of glutathione (GSH), nitrosoadducts of Mb(III) react with it to give nitrosoglutathione, whose concentration has been determined with an apparatus based on a specific and sensitive solid-state amperometric gas sensor. The reaction constant between the adduct and glutathione, kGSH = (47 +/- 1) M(-1) x s(-1), obtained by UV-Vis spectroscopy kinetic measurements, is about one-eighth of the constant with OH- determined by other authors. We can explain this fact with the higher nucleophilicity of OH- compared to GSH, due to the bulkiness and charge of the species. It is known that the formation of nitrosothiols starting from nitrite or NO (nitrogen monoxide) and glutathione, in the absence of oxygen, is impossible. Thus, from a biological point of view, it is important to point out that GSH reacts with NO in the presence of ferrimyoglobin, even at physiological pH, to form nitrosoglutathione. PMID- 11485378 TI - Effects of S-nitroso-N-acetyl-penicillamine administration on glucose tolerance and plasma levels of insulin and glucagon in the dog. AB - It has been suggested that nitric oxide (NO, nitrogen monoxide) is a regulator of carbohydrate metabolism in skeletal muscle. The present study was undertaken to investigate the acute effects of the nitric oxide donor S-nitroso-N acetylpenicillamine (SNAP) on blood glucose levels and on the gluco-regulatory hormones insulin and glucagon in healthy dogs. The acute effects of SNAP on mean arterial pressure and heart rate were also investigated. The drug was administered intravenously and the pre- and postprandial blood glucose, plasma insulin, and glucagon concentrations were determined at half-hour time intervals postadministration after a glucose challenge. The plasma nitrate and nitrite concentrations were measured and taken as the biochemical markers of in vivo NO formation. The oral glucose tolerance test revealed an impaired glucose tolerance in SNAP-treated dogs as reflected by the area under the glucose curve, 1150.50 +/ 63.00 mmol x 150 min and 1355.25 +/- 102.01 mmol/L x 150 min in dogs treated with 10 and 20 mg/kg of SNAP, respectively, compared with 860.25 +/- 60.68 mmol/L x 150 min in captopril-treated controls (P < 0.05). The 2-h blood glucose concentration in dogs treated with 20 mg/kg body wt of SNAP was 9.17 +/- 1.10 mmol/L compared with 5.59 +/- 0.26 mmol/L for captopril-treated controls (P = 0.015). The oral glucose tolerance test also confirmed an impaired insulin secretion in the SNAP-treated dogs. While the plasma insulin concentration increased gradually in the captopril-treated controls to a peak value of 39.50 +/ 2.55 microIU/ml, 1.5 h after a glucose challenge there was a decrease in the plasma insulin concentration in SNAP-treated dogs to a low value of 20.67 +/- 0.88 microIU/ml (P = 0.006). In contrast, there were no significant differences in plasma glucagon concentration in SNAP-treated dogs and captopril-treated dogs at any time points. Using the Griess reaction, we found that there was a 27-95% increase in plasma nitrate/nitrite concentration on administration of SNAP. The sustained hyperglycemic effect observed in SNAP-treated dogs was accompanied by a marginal decrease in the mean arterial blood pressure and a significant increase in heart rate (P < 0.05). We conclude that acute administration of SNAP in the oral glucose tolerance test releases NO that modulates the parameters of carbohydrate metabolism. PMID- 11485379 TI - Oxidation of the ketoxime acetoxime to nitric oxide by oxygen radical-generating systems. AB - Ketoximes undergo a cytochrome P450-catalyzed oxidation to nitric oxide and ketones in liver microsomes. In addition, nitric oxide synthase (NOS) can catalyze the oxidative denitration of the >C=N-OH group of amidoximes. The objective of this work was to characterize the oxidation of a ketoxime (acetoxime) and to assess the ability of NOS to catalyze the generation of nitric oxide/nitrogen monoxide (*NO) from acetoxime. Acetoxime was oxidized to NO2- (and NO3-) by microsomes enriched with several P450 isoforms, including CYP2E1, CYP1A1, and CYP2B1. Nitric oxide was identified as an intermediate in the overall reaction. Superoxide dismutase and catalase significantly inhibited the reaction. Exogenous iron increased the microsomal generation of NO2- from acetoxime, while metal chelators (desferrioxamine, EDTA, DTPA) inhibited it. A Fenton-like system (Fe2+ plus H2O2, pH 7.4) consumed acetoxime with production of NO2- and NO3-, whereas oxidation by superoxide or by H2O2 was inefficient. The results presented suggest a role for hydroxyl radical-like oxidants in the oxidation of acetoxime to nitric oxide. O-Acetylacetoxime and O-tert-butylacetoxime were not oxidized by a Fenton system or by liver microsomes to any significant extent. Formation of the 5,5'-dimethyl-1-pyrroline-N-oxide/. OH adduct by a Fenton system was significantly inhibited by acetoxime, while O-acetylacetoxime and O-tert butylacetoxime were inactive. These results suggest that the. OH-dependent oxidation of acetoxime initially proceeds via abstraction of a hydrogen atom from its hydroxyl group, as opposed to the oxidation of its >C=N- function. HepG2 cells with low levels of expression of P450 did not significantly produce NO2- from acetoxime, while HepG2 cells expressing CYP2E1 did, and this generation was blocked by a CYP2E1 inhibitor. Acetoxime was inactive either as a substrate or as an inhibitor of iNOS activity. These results indicate that reactive oxygen species play a key role in the oxidation of acetoxime to. NO by liver microsomes by a mechanism involving H abstraction from the OH moiety by hydroxyl radical like oxidants and suggest the possibility that acetoxime may be an effective producer of. NO primarily in the liver by a pathway independent of NOS. PMID- 11485380 TI - Hard metal-induced disease: effects of metal cations in vitro on guinea pig isolated airways. AB - Inhalation of dust from hard metal (HM), a mixture of tungsten carbide, cobalt, and other metals, can cause interstitial alveolitis, fibrosis, and asthma in the workplace. Some effects of HM could occur after the metals dissolve in the lung. We examined whether chloride salts of metals in HM alloys can elicit responses or modify reactivity to methacholine (MCh) or responses to electric field stimulation (EFS) in guinea pig tracheal strips. In unstimulated strips, Co(2+), Cd(2+), and Ni(2+) evoked contractions (>3 x 10(-6) M), while Ta(5+), Zn(2+), Cr(2+), and Cr(3+) caused weak relaxations (>10(-5) M). In strips contracted with MCh (3 x 10(-7) M), Co(2+) and Ni(2+) also caused relaxation in lower concentrations while the other metals caused weak relaxation only in high concentrations (>10(-4) M). The metals were generally without effect on reactivity to MCh, except that Cd(2+) inhibited and Ni(2+) potentiated some responses. The effects of selected metals (10(-6) M; Cr(3+), Ni(2+), Cd(2+), and Co(2+)) on EFS-induced contractile and relaxant responses were examined (+/-MCh; +/-10(-6) M indomethacin (Indo), 30 min). No metal had any effect on the excitatory nonadrenergic, noncholinergic-mediated contraction phase. Cd(2+) and Ni(2+) inhibited cholinergically mediated contractions of unstimulated strips (+Indo), whereas Cr(3+) both inhibited (-MCh, -Indo) and potentiated (-Indo,+MCh; +Indo, +MCh) contractile responses. Cr(3+) was the only metal to inhibit the inhibitory nonadrenergic, noncholinergic-mediated relaxation phase (+/-MCh; Indo). Co(2+) had no effect at all. The results suggest that smooth muscle tone and nerves in the airways could be targets of cationic metals after they dissolve in the lung. PMID- 11485381 TI - Intracellular glutathione regulates taurocholate transport in HepG2 cells. AB - The hepatic organic anion transporter 1, Oatp1, was recently demonstrated to function as a GSH exchanger, indicating that hepatic uptake of drugs and xenobiotics may be sensitive to intracellular GSH levels. The present study characterized taurocholate uptake and efflux mechanisms in HepG2 cells and the effects of intracellular GSH on these transport processes. Taurocholate uptake into HepG2 cells was Na(+)-independent, saturable ( K(m) = 82 +/- 16 microM), and was cis-inhibited by bromosulfophthalein and some bile acids. Intracellular GSH depletion inhibited 3H-taurocholate uptake, and, conversely, the release of GSH from HepG2 cells was stimulated in the presence of extracellular taurocholate and other bile acids, consistent with a role for intracellular GSH in stimulating organic anion uptake. Interestingly, efflux of 3H-taurocholate from HepG2 cells was also sensitive to intracellular GSH concentration: efflux was inhibited in cells with lower intracellular GSH and stimulated in cells with higher GSH. RT PCR analysis revealed that OATP-A, OATP-D, OATP-E, OATP-8, MRP1, MRP2, and MRP3 are expressed in HepG2 cells but that their expression is not altered by the maneuvers used to lower or raise intracellular GSH. These results provide direct evidence that intracellular GSH levels modulate both uptake and efflux of taurocholate and suggest that GSH plays a regulatory role in the hepatobiliary transport of potentially toxic organic compounds. PMID- 11485382 TI - In utero/lactational 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure impairs molar tooth development in rats. AB - Developmental defects caused by dioxins are causing increasing concern since they occur at low dose levels and are usually permanent. In this study we examined the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at low in utero/lactational exposure levels on rat tooth development in three rat lines, denoted A, B, and C, that differ in their TCDD sensitivity and aryl hydrocarbon receptor structure. These rat lines are derived from TCDD-resistant Han/Wistar (Kuopio) and TCDD sensitive Long-Evans (Turku/AB) rats by selective breeding. The main target teeth were the third molars, since their development spans from the perinatal period to about 6 weeks after birth. Pregnant dams were exposed to 0.03-1 microg/kg TCDD on gestation day 15. Pups exposed in utero and lactationally were euthanized at the age of 5 or 10 weeks and the jaws were examined. The eruption of the third molar was observed by stereomicroscopy and the jaws were further radiographed. TCDD at 1 microg/kg completely prevented the development of the third lower molars in 60% of males and 50% of females in the most sensitive rat line, C, while only 6% or less of the pups in the more resistant lines A and B were lacking this target tooth. TCDD exposure also dose-dependently diminished the proportion of third molars erupted at the age of 5 weeks. The size of molars was dose-dependently reduced in all rat lines. The third lower molars were most severely affected, and the reduction was significant already at 0.03 microg/kg in line A and at 0.1 microg/kg in lines B and C. The results indicate that impaired tooth development is one of the most sensitive endpoints of TCDD-induced toxicity. PMID- 11485383 TI - Dexamethasone prevents acute cadmium-induced hepatic injury but exacerbates kidney dysfunction in rabbits. AB - Cadmium is a potent hepatotoxicant for which neither effective preventive methods nor the mechanism of toxicity has been established. We investigated the preventive effect of dexamethasone against cadmium toxicity on cadmium-induced liver injury in rabbits. Pretreatment with dexamethasone at 1 mg/kg increased the rate of survival in rabbits administered 2.5 mg/kg iv cadmium. Cadmium induced acute severe liver injury characterized by hepatocellular necrosis, infiltration by inflammatory cells, and increases of plasma GOT, GPT, LDH, and LDH5. Dexamethasone mitigated the acute hepatotoxic effect of cadmium, but exacerbated cadmium-induced kidney dysfunction, with destruction of renal tubular cells and increases in excretion of protein, glucose, and amino acids into urine. The cadmium concentration in liver and kidney of rabbits administered cadmium was not changed by dexamethasone pretreatment. Although metallothionein mRNA expression induced by cadmium was not affected by dexamethasone in liver or kidney, cadmium induced metallothionein protein production was augmented at the early phase in liver and decreased at the later phase in kidney. Neutrophilia observed after cadmium administration was enhanced initially by dexamethasone pretreatment. These results indicate that dexamethasone pretreatment potently prevented cadmium induced liver injury, but exacerbated renal tubular dysfunction. PMID- 11485384 TI - Lead exposure delays the differentiation of oligodendroglial progenitors in vitro. AB - Lead (Pb) is an environmental neurotoxicant that can cause hypo- and demyelination. Oligodendrocytes (OLs), the myelin-forming cells in the central nervous system, may be a possible target for Pb toxicity. The present study describes the effect of Pb on the maturation of rat OL progenitor (OP) cells and the developmental expression of myelin-specific galactolipids. Dose-response studies showed that OP cultures were more sensitive to Pb than mature OLs. Pb delayed the differentiation of OL progenitors, as demonstrated by cell morphology and immunostaining with a panel of stage-specific differentiation markers. Pb given prior to and during differentiation caused a decrease in the biosynthesis of galactolipids in both undifferentiated and differentiated OLs, as detected by metabolic radiolabeling with 3H-D-galactose. While the ratios of galacto/gluco cerebrosides, hydroxy fatty acid/nonhydroxy fatty acid galactolipids, and galactocerebrosides/sulfatides increased in control cultures during cell differentiation, Pb treatment prevented these changes. The results suggest that chronic Pb exposure may impact brain development by interfering with the timely developmental maturation of OL progenitors. PMID- 11485385 TI - Differential induction of apoptosis and MAP kinase signaling by mitochondrial toxicants in drug-sensitive compared to drug-resistant B-lineage lymphoid cell lines. AB - A panel of human B-lineage lymphoma cell lines differing in cancer drug resistance status and Bcl-2/Bax expression were used to study the contribution of mitochondrial-based perturbations and regulation in differential induction of apoptosis. Mitochondrial dysfunction was induced in cells by the uncoupler carbonyl cyanide m-chlorophenylhydrazone (mClCCP) and the respiratory chain inhibitor antimycin A. Cells were then assayed for early changes in MAP kinase signaling and subsequent induction of apoptosis. The cancer drug-resistant cell lines EW36 and CA46, overexpressing Bcl-2 and deficient in Bax, respectively, were both resistant to mitochondrial toxicant-induced cleavage of poly(ADP ribose) polymerase (PARP) and morphologically detectable apoptotic cell death. In contrast, cancer drug-sensitive ST486 cell line, with low Bcl-2 expression, was sensitive to PARP cleavage and apoptosis engagement. Interestingly, mClCCP induced twofold more apoptosis than antimycin A in the ST486 cells. Exposure to the mitochondrial toxicants resulted in the early and preferential activation of the ERK and p38 MAP kinase pathways in only the drug-sensitive ST486 cell line, with mClCCP more potent than antimycin A. Specific inhibition of the p38 pathway augmented baseline and mClCCP-induced apoptosis. These results show that multi drug-resistant and -sensitive B-lineage cells are also resistant and sensitive to compounds inducing mitochondrial dysfunction. The differential sensitivity to mitochondrial toxicant effects involved regulation by MAP kinases, since ERK and p38 were found to be preferentially activated only in the drug-sensitive B lineage cells. Modulation of the p38 signaling pathway altered the sensitivity of cells to mitochondrial stress and may play a more general role in regulating the sensitivity of B-lineage cells to drugs and environmental toxicants. PMID- 11485386 TI - Mercuric chloride enhances immunoglobulin E-dependent mediator release from human basophils. AB - Mercuric chloride (HgCl2) is an industrial agent known to cause autoimmune disorders and induce IgE synthesis, which plays a crucial role in the manifestation of allergic diseases. In rodents, the immunomodulatory effects of HgCl2 have been shown to involve the enhancement of mast cell-derived IL-4 secretion, which facilitates both Th2-lymphocyte development and IgE production. In humans, rapid allergen-dependent release of IL-4 and the related cytokine IL 13 from histamine-containing cells occurs primarily in basophils, along with other proinflammatory mediators such as histamine and LTC4. In this study, we therefore investigated the effects of HgCl2 on the release of the above basophil mediators, either due to the compound alone or in conjunction with IgE-dependent stimulation. HgCl2 (10(-9) to 10(-6) M) did not induce mediator secretion alone but significantly enhanced the release of histamine, LTC4, IL-4, and IL-13 caused by anti-IgE. Higher concentrations of HgCl2 (10(-5) to 10(-3) M) strikingly reduced cell viability; however, toxicity varied depending on cell density and incubation time. Removal of HgCl2 following a short incubation with basophils did not reverse the potentiating effects on basophil mediator secretion to anti-IgE and the concentration of free mercury in the supernatants significantly diminished by up to 20% after incubation with the cells, indicating irreversible Hg binding to cells. By upregulating IgE-dependent human basophil mediator release, our results clearly indicate that HgCl2 potentially exacerbates allergic disorders and promotes a Th2-cytokine profile. PMID- 11485387 TI - Marijuana smoke and Delta(9)-tetrahydrocannabinol promote necrotic cell death but inhibit Fas-mediated apoptosis. AB - Marijuana smoke shares many components in common with tobacco smoke except for the presence of Delta(9)-tetrahydrocannabinol (Delta(9)-THC), the psychotropic compound found only in Cannibis sativa. Delta(9)-THC has been shown to potentiate smoke-induced oxidative stress and necrotic cell death. In the present study, our objective was to determine the effects of Delta(9)-THC on the balance between Fas induced apoptosis and necrosis in A549 lung tumor cells. We found that Fas induced activation of caspase-3 was inhibited by whole smoke from both tobacco and marijuana cigarettes. Gas-phase smoke, which generates high levels of intracellular reactive oxygen species, had no effect on caspase-3 activity. However, particulate-phase smoke (tar) was a potent inhibitor of Fas-induced caspase-3 activity, with marijuana tar being more potent than either tobacco or placebo marijuana tar (lacking Delta(9)-THC). Delta(9)-THC also inhibited Fas induced caspase-3 activity in A549 cells. In contrast, no inhibition was observed when Delta(9)-THC was incubated with activated caspase-3 enzyme, suggesting that Delta(9)-THC acts on the cell pathway(s) leading to caspase-3 activation and not directly on enzyme function. Flow cytometry was used to measure the percentage of cells undergoing apoptosis (staining for annexin V) versus necrosis (staining for propidium iodide) and confirmed that both marijuana tar extract and synthetic Delta(9)-THC inhibit Fas-induced apoptosis while promoting necrosis. These observations suggest that the Delta(9)-THC contained in marijuana smoke disrupts elements of the apoptotic pathway, thereby shifting the balance between apoptotic and necrotic cell death. This shift may affect both the carcinogenic and immunologic consequences of marijuana smoke exposure. PMID- 11485388 TI - Proteolytic loss of bcl-x(L) in FL5.12 Cells undergoing apoptosis induced by MK886. AB - Apoptosis induced in the IL3-dependent murine pro-B lymphocytic (FL5.12) cell line by the 5-lipoxygenase activating protein inhibitor MK886 is accompanied by the rapid loss of the anti-apoptotic bcl-x(L) and bcl-2, but not the proapoptotic bax proteins (Datta et al., J. Biol. Chem. 273, 28163-28169, 1998). Since several reports indicate important roles for noncaspase proteases in apoptosis, the participation of lysosomes, as well as serine, cysteine, or aspartic acid proteases, in the effects of MK886 were investigated. Consistent with the involvement of various proteases, lysosomal degranulation was evident, as observed by a decrease in acridine orange fluorescence at 2 h and an increase in cytosolic beta-hexosaminidase activity at 4 h after treating FL5.12 cells with 10 microM MK886. The disappearance of bcl-x(L) from FL5.12 cells upon MK886 treatment was prevented in a dose-dependent manner by pretreatment with leupeptin, pepstatin, phenylmethylsulfonyl fluoride, or the broad-spectrum caspase inhibitor Boc-D-FMK. Each of the noncaspase protease inhibitors partially inhibited MK886-induced apoptosis as measured by phosphatidylserine externalization and DNA fragmentation. The noncaspase inhibitors also blocked about half of the increase in caspase-3-like activity. Boc-D-FMK completely inhibited this enzyme and prevented apoptosis. None of the inhibitors were able to directly inhibit activated caspase-3 in cell lysates, suggesting their effects were upstream of caspase activation. These observations suggest the involvement of various proteases, possibly originating from lysosomes, upstream of active caspase-3, in the loss of bcl-x(L) protein and in the signaling pathway of MK886 induced apoptosis in FL5.12 cells. This pathway may be unique to MK886 since these same protease inhibitors had only minimal effects on etoposide-induced apoptosis and the accompanying moderate loss of bcl-x(L) in FL5.12 cells. PMID- 11485389 TI - Determination of trivalent methylated arsenicals in biological matrices. AB - The enzymatically catalyzed oxidative methylation of As yields methylated arsenicals that contain pentavalent As (As(V)). Because trivalent As (As(III)) is the favored substrate for this methyltransferase, methylated arsenicals containing As(V) are reduced to trivalency in cells. Methylated arsenicals that contain As(III) are extremely potent inhibitors of NADPH-dependent flavoprotein oxidoreductases and potent cytotoxins in many cell types. Therefore, the formation of methylated arsenicals that contain As(III) may be properly regarded as an activation step, rather than a means of detoxification. Recognition of the role of methylated arsenicals that contain As(III) in the toxicity and metabolism of As emphasizes the need for analytical methods to detect and quantify these species in biological samples. Hence, a method was developed to exploit pH dependent differences in the generation of arsines from inorganic and methylated arsenicals that contain either As(V) or As(III). Reduction with borohydride at pH 6 generated arsines from inorganic As(III), methyl As(III), and dimethyl As(III), but not from inorganic As(V), methyl As(V), and dimethyl As(V). Reduction with borohydride at pH 2 or lower generated arsines from arsenicals that contained either As(V) or As(III). Arsines are trapped in a liquid nitrogen-cooled gas chromatographic trap, which is subsequently warmed to allow separation of the hydrides by their boiling points. Atomic absorption spectrophotometry is used to detect and quantify the arsines. The detection limits (ng As ml(-1)) for inorganic As(III), methyl As(III), and dimethyl As(III) are 1.1, 1.2, and 6.5, respectively. This method has been applied to the analysis of arsenicals in water, human urine, and cultured cells. Both methyl As(III) and dimethyl As(III) are detected in urine samples from individuals who chronically consumed inorganic As-contaminated water and in human cells exposed in vitro to inorganic As(III). The reliable quantitation of inorganic and methylated arsenicals that contain As(III) in biological samples will aid the study of the toxicity of these species and may provide a new biomarker of the effects of chronic exposure to As. PMID- 11485390 TI - Activation of the activator protein-1 by the peroxisome proliferator clofibric acid in rat H4IIEC3 hepatoma cells. AB - Clofibric acid (CA), a potent peroxisome proliferator (PP), has been shown to cause tumor formation in rat liver. The precise mechanism of action of PPs remains largely unknown. However, it has been proposed that they act by increasing reactive oxygen species (ROS), leading to a cellular oxidative stress. In the present study, we have investigated the effect of CA on the activator protein-1 (AP-1) expression in PP-responsive H4IIEC3 rat hepatoma cells. Electrophoretic mobility shift assays demonstrated that AP-1 activation was induced in cells treated with CA for 24 h at all concentrations of the fibrate. This activation was prolonged up to 48 h. Using transfection experiments with H4IIEC3 cells, we found that CA induced the expression of a reporter gene driven by AP-1 and that of the glutathione S-transferase P target gene. By supershift experiments, jun and fos proteins were identified as components of the CA activated AP-1 complexes. Western blot analyses revealed that the induction of the AP-1 activity was not dependent to an increase in the levels of jun and fos proteins. Cotreatment of H4IIEC3 cells with CA and the antioxidant N acetylcysteine or calphostin C, a specific inhibitor of protein kinase C (PKC), blocked the AP-1 activation and the expression of the AP-1-driven luciferase reporter gene. These results demonstrate that CA activates AP-1 in H4IIEC3 cells and that this induction is mediated via ROS and PKC. PMID- 11485391 TI - Keratinocyte differentiation marker suppression by arsenic: mediation by AP1 response elements and antagonism by tetradecanoylphorbol acetate. AB - Culture models of target cells are anticipated to help elucidate the mechanism by which inorganic arsenic acts as a carcinogen in humans. Present work characterizes the response of human keratinocytes, a target cell type, to arsenic suppression of their differentiation program. Four representative differentiation marker mRNAs (involucrin, keratinocyte transglutaminase, small proline-rich protein 1, and filaggrin) were suppressed by both arsenate and arsenite in normal, spontaneously immortalized (premalignant), and malignant keratinocytes with EC50 values in the low micromolar range. The suppression was almost completely reversed 9 days after removal of arsenate from the culture medium. In the case of the involucrin gene, suppression was mediated primarily by two functional AP1 response elements in the gene promoter. Both glucocorticoid and serum stimulation of differentiation occurred to a similar extent in the presence and absence of arsenic, indicating neither stimulation was a specific target of arsenic action and neither agent could overcome arsenic suppression. In contrast, 12-O-tetradecanoylphorbol-13-acetate prevented the suppression of keratinocyte transglutaminase, suggesting that arsenic acts upstream of protein kinase C. PMID- 11485394 TI - Beet necrotic yellow vein virus particles localize to mitochondria during infection. AB - Fluorescent beet necrotic yellow vein virus (BNYVV) particles were produced by replacing part of the readthrough domain of the minor coat protein P75 with the green fluorescent protein (GFP). The recombinant virus was functional in plants and P75-GFP was incorporated at one end of the rod-shaped virions. Laser scanning confocal microscopy and transmission electron microscopy showed that virus-like particles, almost certainly authentic BNYVV virions, localized to the cytoplasmic surface of mitochondria at early times postinfection but relocated at later times to semiordered clusters in the cytoplasm. This is the first report of specific targeting of plant virus particles to the mitochondria in vivo. PMID- 11485393 TI - The interferon-inducible 204 gene is transcriptionally activated by mouse cytomegalovirus and is required for its replication. AB - Infection of cells with viable or UV-inactivated murine cytomegalovirus (MCMV) increased the IFN-inducible 204 gene at both the mRNA and the protein levels. The activity of a reporter gene driven by the mouse Ifi204 promoter induced following virus infection showed that this increase was due to transcriptional activation. Moreover, FACS analysis of infected mouse embryo fibroblasts (MEF) stably transfected with a p204-dominant-negative mutant (p204dmMEF) revealed that they do not accumulate at the G1/S border in the same way as infected MEF transfected with the empty vector (neoMEF). MCMV DNA synthesis is significantly delayed (144 h in p204dmMEF vs 72 h in neoMEF), due to retarded expression of viral genes, namely, IE1 and DNA polymerase, as shown by Western blot comparison of p204dmMEF and neoMEF extracts. These results demonstrate that MCMV may exploit the Ifi204 gene to regulate the cell cycle and enhance its DNA synthesis. PMID- 11485395 TI - Characterization of pseudotype VSV possessing HCV envelope proteins. AB - The genome of hepatitis C virus (HCV) encodes two envelope glycoproteins (E1 and E2), which are thought to be responsible for receptor binding and membrane fusion resulting in virus penetration. To investigate cell surface determinants important for HCV infection, we used a recombinant vesicular stomatitis virus (VSV) in which the glycoprotein gene was replaced with a reporter gene encoding green fluorescent protein (GFP) and produced HCV-VSV pseudotypes possessing chimeric HCV E1 or E2 glycoproteins, either individually or together. The infectivity of the pseudotypes was determined by quantifying the number of cells expressing the GFP reporter gene. Pseudotypes that contained both of the chimeric E1 and E2 proteins exhibited 10--20 times higher infectivity on HepG2 cells than the viruses possessing either of the glycoproteins individually. These results indicated that both E1 and E2 envelope proteins are required for maximal infection by HCV. The infectivity of the pseudotype virus was not neutralized by anti-VSV polyclonal antibodies. Bovine lactoferrin specifically inhibited the infection of the pseudotype virus. Treatment of HepG2 cells with Pronase, heparinase, and heparitinase but not with phospholipase C and sodium periodate reduced the infectivity. Therefore, cell surface proteins and some glycosaminoglycans play an important role in binding or entry of HCV into susceptible cells. The pseudotype VSV possessing the chimeric HCV glycoproteins might offer an efficient tool for future research on cellular receptors for HCV and for the development of prophylactics and therapeutics for hepatitis C. PMID- 11485396 TI - Influence of the small leader exons 2 and 3 on human immunodeficiency virus type 1 gene expression. AB - The human immunodeficiency virus type 1 (HIV-1) uses an elaborate alternative splicing pattern for the generation of both the 1.8-kb as well as the 4-kb classes of mRNA. An additional diversity of transcripts in both classes is created by the optional inclusion of the small exons 2 and 3 in the leader sequence. To analyze a possible influence of these leader exons on HIV-1 gene expression, several series of expression vectors with different leaders were constructed, expressing either Rev and Env or a heterologous coding sequence, i.e., the chloramphenicol acetyl transferase (CAT) ORF. Transfection experiments of HeLa-T4(+) cells revealed for all series of constructs that mRNA as well as protein expression was stimulated by the presence of exon 2 and reduced by exon 3. The function of the leader exons 2 and 3 is neither dependent on the regulatory proteins Tat or Rev nor on viral coding sequences. Neither transcription rates nor stability of polyadenylated RNAs were found to be responsible for the different levels of steady-state mRNA. When either exon 2 or 3 was inserted into a heterologous intron, processing of the primary transcripts generated identical mRNA species while maintaining the differences in exon 2/3 dependent mRNA steady-state levels. These results may be explained by exon specific nuclear RNA degradation rates, as also indicated by results from an in vitro degradation assay using a HeLa nuclear extract. PMID- 11485397 TI - Crystallization of Brome mosaic virus and T = 1 Brome mosaic virus particles following a structural transition. AB - Brome mosaic virus (BMV), a T = 3 icosahedral plant virus, can be dissociated into coat protein subunits and subunit oligomers at pH 7.5 in the presence of concentrated salts. We have found that during the course of this treatment the coat protein subunits are cleaved, presumably by plant cell proteases still present in the preparation, between amino acids 35 and 36. The truncated protein subunits will then reorganize into T = 1 icosahedral particles and can be crystallized from sodium malonate. Quasi elastic light scattering and atomic force microscopy results suggest that the transition from T = 3 to T = 1 particles can occur by separate pathways, dissociation into coat protein subunits and oligomers and reassembly into T = 1 particles, or direct condensation of the T = 3 virions to T = 1 particles with the shedding of hexameric capsomeres. The latter process has been directly visualized using atomic force microscopy. Native T = 3 virions have been crystallized in several different crystal forms, but neither a rhombohedral form nor either of two orthorhombic forms diffract beyond about 3.4 A. Tetragonal crystals of the T = 1 particles, however, diffract to at least 2.5 A resolution. Evidence suggests that the T = 1 particles are more structurally uniform and ordered than are native T = 3 virions. A variety of anomalous virus particles having diverse sizes have been visualized in preparations of BMV used for crystallization. In some cases these aberrant particles are incorporated into growing crystals where they are frequently responsible for defect formation. PMID- 11485398 TI - Paired DNA three-way junctions as scaffolds for assembling integrase complexes. AB - Early steps of retroviral replication involve reverse transcription of the viral RNA genome and integration of the resulting cDNA copy into a chromosome of the host cell. The initial DNA breaking and joining steps of integration are carried out by the virus-encoded integrase enzyme. Integrases bind specifically to the ends of the unintegrated viral cDNA but nonspecifically to target DNA. Conventional assays in vitro reveal primarily the nonspecific DNA binding mode, complicating studies of integrase--DNA complexes. Here, we report an investigation of unconventional DNA structures useful for positioning integrase at predetermined sites. We find that paired DNA three-way junctions can be used to mimic branched DNAs normally formed as reaction intermediates. The three-way junctions differ from authentic intermediates in the connectivity of the DNAs, which, in contrast to the authentic intermediate, allow formation of stable DNA structures under physiological conditions. Assays in vitro showed that integrase can direct hydrolysis at sequences resembling the viral cDNA ends within the three-way junction, but not on junctions with mutant sequences. Changing the spacing between the paired three-way junctions disrupted the cleavage pattern, emphasizing the importance of the correct DNA scaffold. DNase I footprinting studies revealed protection of specific bases at the terminus of the LTR in the three-way junction complex, but not on control linear DNA, specifying the locations of tight interactions between integrase and DNA. Paired DNA three-way junctions are attractive reagents for structural studies of integrase-DNA complexes. PMID- 11485399 TI - In vitro analysis of an RNA binding site within the N-terminal 30 amino acids of the southern cowpea mosaic virus coat protein. AB - Southern cowpea mosaic virus (SCPMV) is a positive-sense RNA virus with T = 3 icosahedral symmetry. The coat protein (CP) has two domains, the random (R) domain and the shell (S) domain. The R domain is formed by the N-terminal 64 amino acids (aa) and is localized to the interior of the particle where it is expected to interact with the viral RNA. The R domain (aa 1--57) was expressed in Escherichia coli as a recombinant protein (rWTR) containing a nonviral C-terminal extension with two histidine tags. The RNA binding site of the R domain was identified by Northwestern blotting and electrophoretic mobility shift assay (EMSA) using recombinant wild-type and mutant R domain proteins. Deletions within the R domain revealed that the RNA binding site is localized to its N-terminal 30 aa. RNA binding by this element was found to be nonspecific with regard to RNA sequence and was sensitive to high salt concentrations, suggesting that electrostatic interactions are important for RNA binding by the R domain. The RNA binding site includes 11 basic residues, eight of which are located in the arginine-rich region between aa 22 and 30. It was demonstrated using alanine substitution mutants that the basic residues of the arginine-rich region but not those present at positions 3, 4, and 7 are necessary for RNA binding. None of the basic residues within the arginine-rich region are specifically required for RNA binding, but the overall charge of the N-terminal 30 aa is important. Proline substitution mutations within the N-terminal 30 aa, and alanine substitutions for prolines at positions 18, 20, and 21, did not affect the RNA binding activity of the R domain. However, it was demonstrated by circular dichroism (CD) that the conformation of the N-terminal 30 aa of the R domain changes from a random coil to an alpha-helix in the presence of 50% trifluoroethanol (TFE). The possible role for this structural change in RNA binding by the R domain is discussed. PMID- 11485400 TI - Infection with Langat Flavivirus or expression of the envelope protein induces apoptotic cell death. AB - Langat (LGT) flavivirus, derived from infectious full-length cDNA clone 636, was investigated for its apoptotic activities in mouse neuroblastoma (Neuro-2a) and simian kidney (Vero and LLC-MK(2)) cells. The hallmark of apoptosis, cleavage of cellular DNA, was observed 48 h after infection of Vero, LLC-MK(2), and Neuro-2a cells by electrophoresis analysis. Apoptosis in infected cells was also confirmed by TUNEL assay. LGT-infected Neuro-2a cells showed an increase in caspase-3-like protease (DEVDase) activity. Expression of the major envelope glycoprotein (E) alone reduced cell viability in both Vero and Neuro-2a cells, and the baculovirus P35 protein, which inhibits multiple caspases, completely blocked this effect. Cleavage of cellular DNA was observed in E gene-transfected Vero cells by TUNEL assay. Expression of E protein or caspase-9 resulted in activation of caspase-3 like proteases in Neuro-2a cells. The caspase-3-like protease specific inhibitor, Ac-DEVD-CHO peptide, partially inhibited E protein- or caspase-9-induced apoptosis in Neuro-2a cells. These observations indicate that infection of cells with LGT virus or expression of LGT virus E protein induces apoptosis through a caspase-3-like protease pathway. PMID- 11485401 TI - UDP-galactose transporter is required for Theiler's virus entry into mammalian cells. AB - Theiler's murine encephalomyelitis viruses (TMEV) are divided into two groups: high-neurovirulence strains, such as GDVII, cause fatal encephalitis, while low neurovirulence strains, such as BeAn and DA, cause persistent infection and demyelination in mice. Cell surface sialic acid is bound by the low neurovirulence DA and BeAn viruses, but not by the high-neurovirulence GDVII virus. We have identified a clone from a BHK-21 cell cDNA library that mediates TMEV entry and infection by viruses of both TMEV groups in a receptor-negative BHK-21 cell variant (R26). The sequence of this clone is 96.4% identical to the human UDP-galactose transporter (UGT), which belongs to a family of nucleotide sugar transporter proteins involved in the biosynthesis of complex carbohydrate structures in the trans-Golgi network. UGT mRNA from R26 cells was found to have a 490-nucleotide deletion involving the C-terminal amino acids 255 to 392 and 81 nucleotides of the 3' noncoding region. These results suggest two possibilities by which UGT may mediate TMEV entry and infection. The most likely one relates to the transporter function of adding galactose to another receptor protein. This possibility suggests the requirement for a specific glycoprotein interaction for GDVII virus cell binding and entry, e.g., galactose for GDVII and sialic acid for BeAn. Alternatively, UGT might be a TMEV receptor itself, acting via UGT cycling to the cell surface. PMID- 11485402 TI - Genetic identification and characterization of limestone canyon virus, a unique Peromyscus-borne hantavirus. AB - Hantaviruses, family Bunyaviridae, are rodent-borne RNA viruses that can cause hantavirus pulmonary syndrome (HPS) in various regions of the Americas. A coevolutionary relationship exists between hantaviruses and their specific rodent reservoir hosts; the phylogeny of the viruses generally matches that of the rodents. There are several Peromyscus-borne hantaviruses, including Sin Nombre virus, the most common cause of HPS in North America. This report describes the genetic detection and characterization of a newly discovered Peromyscus boylii borne virus, Limestone Canyon (LSC) virus, the most divergent member of the Peromyscus-borne hantaviruses to date. Analysis of a 1209-nucleotide region of the S segment of LSC virus showed it to be more closely related to hantaviruses found in harvest mice (Reithrodontomys megalotis and R. mexicanus) than to other Peromyscus-associated hantaviruses (Sin Nombre, New York, and Monongahela). Phylogenetic analysis of virtually the entire M genome segment (3489 nucleotides) of LSC virus revealed a similar picture in which LSC virus was found to be very distinct from other Peromyscus-associated viruses, but its exact relationship to the other Peromyscus-borne and the Reithrodontomys-borne viruses was not resolved. These results indicate that hantavirus host species-jumping events can occur by which a hantavirus may switch to, and become established in, a rodent host belonging to a different genus. P. boylii are present throughout the southwestern United States and central Mexico. More extensive screening of HPS patients by using RT-PCR assays will be necessary to determine if LSC virus can cause human disease. PMID- 11485403 TI - Genome sequence determinations and analyses of novel circoviruses from goose and pigeon. AB - The genomes of novel circoviruses from goose and pigeon, which were isolated using degenerate primer and inverse primer PCR methods, were cloned and sequenced. Comparative nucleotide (nt) sequence analyses showed that the goose circovirus (GCV) and pigeon circovirus (PiCV) possessed genomes which were 1821 and 2037 or 2036 nt, respectively, and which had features in common with the genomes of porcine circoviruses types 1 and 2 (PCV1, PCV2) and psittacine beak and feather disease virus (BFDV), such that they can now be assigned to the genus Circovirus of the family Circoviridae. Common features include the possession of (i) a potential stem-loop/nonanucleotide motif with which the initiation of rolling circle replication of the virus DNA is associated; (ii) two major ORFs, located on the virus (V1 ORF) and complementary (C1 ORF) strands, which encode the replication-associated protein (Rep) and capsid protein, respectively; (iii) high levels of amino acid identity (41.2--58.2%) shared with other circovirus Rep proteins; and (iv) direct/inverted repeat sequences within the putative intergenic region. On the basis of nt and amino acid sequence identities, GCV is substantially less closely related to BFDV than PiCV is to BFDV. PMID- 11485404 TI - Umbravirus gene expression helps potato leafroll virus to invade mesophyll tissues and to be transmitted mechanically between plants. AB - Potato leafroll virus (PLRV) was mechanically transmissible when inocula also contained the umbravirus Pea enation mosaic virus-2 (PEMV-2). In plants infected with PLRV and PEMV-2, PLRV accumulated in clusters of mesophyll cells in both inoculated and systemically infected leaves. No transmissions were obtained by coinoculation with Potato virus Y, Potato virus X (PVX), Tobacco mosaic virus, or Cucumber mosaic virus (CMV), although PLRV was transmissible from mixtures with CMV(ORF4) (a recombinant that contained the movement protein (MP) gene of the umbravirus Groundnut rosette virus (GRV) in place of the CMV MP gene). In contrast, neither a recombinant PVX that expressed GRV MP nor a mutant of CMV(ORF4), in which the CMV 2b gene was untranslatable, was able to help PLRV transmission. Possibly both a cell-to-cell movement function and counterdefense mechanisms such as those that block posttranscriptional gene silencing are involved in movement of PLRV within plants and its mechanical transmission between plants. PMID- 11485405 TI - Subcellular targeting of the coat protein of African cassava mosaic geminivirus. AB - The coat protein (CP) of geminiviruses is involved in a number of processes during the life cycle of the virus. The predominant function is encapsidation of single-stranded DNA and formation of the virus particle to protect viral DNA during transmission. The CP of monopartite geminiviruses is absolutely essential for virus movement, whereas CP mutants of bipartite geminiviruses are able to infect some host plants systemically, indicating an involvement of the CP in host specificity. During the life cycle of geminiviruses, the viral DNA enters the nucleus of the infected cell where virus replication, transcription, and encapsidation occur. For systemic infection, the virus moves cell-to-cell from the site of inoculation to vascular tissue and via phloem to other plant tissues. To move, viral DNA has to shuttle in and out of the nucleus and through plasmodesmata. Parts of the bipartite African cassava mosaic virus (ACMV) CP were fused with green fluorescent protein (GFP) or beta-glucuronidase (GUS). CP domains were identified that mediate both nuclear import and export, as well as targeting of CP-fusion proteins to the cell periphery. These results indicate that domains of the CP facilitate several aspects of geminivirus movement, including nuclear import and export and transport of the viral genome to the cell periphery. PMID- 11485406 TI - Vaccine potential of Ebola virus VP24, VP30, VP35, and VP40 proteins. AB - Previous vaccine efforts with Ebola virus Zaire (EBOV-Z) emphasized the potential protective efficacies of immune responses to the surface glycoprotein and the nucleoprotein. To determine whether the VP24, VP30, VP35, and VP40 proteins are also capable of eliciting protective immune responses, these genes were expressed from alphavirus replicons and used to vaccinate BALB/c and C57BL/6 mice. Although all of the VP proteins were capable of inducing protective immune responses, no single VP protein protected both strains of mice tested. VP24, VP30, and VP40 induced protective immune responses in BALB/c mice, whereas C57BL/6 mice survived challenge only after vaccination with VP35. Passive transfer of immune sera to the VP proteins did not protect unvaccinated mice from lethal disease. The demonstration that the VP proteins are capable of eliciting protective immune responses to EBOV-Z indicates that they may be important components of a vaccine designed to protect humans from Ebola hemorrhagic fever. PMID- 11485407 TI - Cytoplasmic localization is important for transcription factor nuclear factor kappa B activation by hepatitis C virus core protein through its amino terminal region. AB - We previously reported that hepatitis C virus core protein (core) activates the transcription factor nuclear factor-kappa B (NF-kappa B) when expressed transiently. In the present study, we investigated the relationship between the NF-kappa B activation capacity and subcellular localization of the core. By changing the subcellular localization of the C-terminally truncated core from the nucleus to the cytoplasm, NF-kappa B was activated. In addition, NF-kappa B activity was augmented by forcing the mutated core to move to the endoplasmic reticulum. It was also suggested that the region from aa 21 to 80 of the core is involved in the activation of NF-kappa B. PMID- 11485408 TI - Viral neuraminidase treatment of dendritic cells enhances antigen-specific CD8(+) T cell proliferation, but does not account for the CD4(+) T cell independence of the CD8(+) T cell response during influenza virus infection. AB - In vitro studies demonstrate that the increased alloreactive T cell response to dendritic cells (DC) that are treated with either live or inactivated influenza virus A/PR/8/34 is due to viral neuraminidase (NA) activity. Since virus-specific cytotoxic T lymphocytes (CTL) play an important role in immunity to heterologous influenza strains, we compared the activation of CD8(+) T cells by untreated and NA-treated DC. Increased CTL activity was induced by NA-treated DC both in vitro and in vivo. Since the generation of CTL in response to influenza virus infection does not require prior "activation" of DC by CD4(+) T cells (as is the case for many antigens), we asked whether NA activity contributed to this unconditional CD8(+) T cell response. This was not the case. Future studies will determine the factors that are responsible for the CD4(+) T-cell-independent influenza virus specific CTL response. PMID- 11485409 TI - HIV-1 NL4-3, but not IIIB, inhibits JAK3/STAT5 activation in CD4(+) T cells. AB - HIV-1 infection leads to T cell dysfunction and apoptosis in vivo and in vitro. The shared common gamma chain of IL-2R and its associated Janus kinase, JAK3, are indispensable for normal T cell function and survival. We have reported that CD4 ligation with HIV gp120 inhibits T cell receptor-induced activation and expression of JAK3. We have also shown that while some strains of HIV-1, such as NL4-3, induce apoptosis of infected CD4(+) T cells, other strains, such as HIV-1 IIIB, do not. Interestingly, we show here that infection of CD4(+) T cells with HIV-1 NL4-3, but not IIIB, inhibited activation and expression of JAK3. NL4-3 infected T cells were unable to upregulate JAK3 expression following stimulation through TCR/CD3. In addition, NL4-3, but not IIIB, inhibited tyrosine phosphorylation and expression of STAT5, a downstream target of JAK3. These data suggest a correlation between apoptosis of HIV-1-infected T cells and inhibition of the JAK3/STAT5 activation pathway. PMID- 11485411 TI - Caprine arthritis encephalitis virus: evidence for a B/D-type assembly pathway in a C-type lentivirus replication. AB - Lentiviruses, among which is caprine arthritis encephalitis virus (CAEV), are known to concomitantly assemble and bud at the plasma membrane of infected cells, in a C-type defined pathway. Electron microscopy analysis of CAEV-infected cells demonstrated viral particles budding at the plasma membrane and into intracellular membrane-surrounded vesicles. Furthermore, nonenveloped immature virus-like particles, resembling intracytoplasmic type-A particles (ICAPs), accumulated within the cytoplasm of those cells. Fractionation on sucrose density gradients of cytoplasmic lysates from CAEV-infected cells revealed that enveloped immature or mature viral particles had a density of 1.16--1.17 g/ml, whereas ICAPs sedimented at a density of 1.2--1.27 g/ml. Endogenous reverse transcriptase activity was only associated with the 1.16--1.17 g/ml density particles despite the presence of viral RNA in both populations. The intracellular enveloped particles were found to be infectious. The CAEV Gag precursor by itself was shown to direct assembly, budding, and release of immature virus-like particles when expressed in goat primary synovial membrane cells using the same pathways of assembly and budding as observed in CAEV-infected cells. These data suggest that CAEV assembly, driven by the Gag precursor, could unusually proceed via two simultaneous pathways characteristic of type-C and type-B/D retroviruses. PMID- 11485410 TI - Disruption of NF-kappa B signaling and chemokine gene activation by retroviral mediated expression of IKK gamma/NEMO mutants. AB - Phosphorylation of I kappa Bs--the cytoplasmic inhibitors of the NF-kappa B transcription factors--is the key event which triggers activation of the NF-kappa B cascade. Signal-mediated phosphorylation of I kappa B alpha is mediated by a multiprotein complex, the I kappa B kinase (IKK) complex, which is composed of at least three identified subunits. Two of these polypeptides, IKK alpha and IKK beta, also known as IKK1 and IKK2, are the catalytic subunits of the kinase complex and phosphorylate I kappa B alpha and I kappa B beta. The third component, NEMO/IKK gamma, does not exhibit kinase activity, but rather constitutes a regulatory subunit. In the present study, C-terminal truncated forms of IKK gamma--Delta C-IKK gamma 306 and Delta C-IKK gamma 261--were stably expressed in the myeloid cell line U937 by retroviral-mediated gene transfer. Overexpression of Delta C-IKK gamma resulted in a reduction in IKK kinase activity in vitro, a subsequent decrease in NF-kappa B DNA binding activity, and inhibition of chemokine gene induction in response to TNFalpha stimulation or paramyxovirus infection. This study demonstrates the efficacy of Delta C-IKK gamma as a repressor of IKK signaling and NF-kappa B activation and suggests a potential gene therapy approach to limit chronic inflammation due to chemokine hyperactivation. PMID- 11485412 TI - Human-herpesvirus-8-encoded K8 protein colocalizes with the promyelocytic leukemia protein (PML) bodies and recruits p53 to the PML bodies. AB - Promyelocytic leukemia protein (PML) bodies are nuclear sites for both input viral genome deposition and immediate-early (IE) gene transcription during infection with certain human DNA viruses, such as human cytomegalovirus (HCMV), herpes simplex virus type 1, and adenovirus. In this study, we showed that the K8 (K-bZIP) protein, an early protein encoded by the human herpesvirus 8 (HHV-8), colocalized with the PML bodies in HHV-8-infected primary effusion lymphoma cells. Cotransfection of two plasmids expressing the K8 protein and green fluorescence protein (GFP)-PML fusion protein into 293T cells revealed that the K8 protein colocalized with PML in cells with high PML expression. Overexpression of the K8 protein in Chinese hamster ovary (CHO) cells with stable GFP-PML expression did not induce the dispersion of the PML bodies, unlike the IE1 protein of HCMV. Transfection of a truncated K8 gene revealed that the leucine zipper domain of the K8 protein was required for the colocalization with PML. We also demonstrated that the K8 protein bound to p53 in vivo and in vitro, and that high expression of the K8 protein caused the accumulation of p53 to the PML bodies in CHO cells, suggesting that the K8 protein functions in the recruitment of p53 to the PML bodies. These data suggest that the K8 protein may be associated with the functional modulation of p53 in the nucleus during the lytic phase of HHV-8. PMID- 11485413 TI - Evidence for the presence of two novel pestivirus species. AB - The genus Pestivirus of the family Flaviviridae comprises four species, namely Bovine viral diarrhea virus-1 (BVDV-1), BVDV-2, Border disease virus (BDV), and Classical swine fever virus (CSFV). Comparative analyses of partial sequences have suggested that pestivirus isolates from giraffe (Giraffe-1) and reindeer (Reindeer-1) are distinct from the established species (Becher et al., Virology 262, 64--71, 1999). In this study, we report the complete genomic sequences of pestivirus strains Giraffe-1 and Reindeer-1. Comparative sequence analyses revealed considerable differences among Giraffe-1, Reindeer-1, and the currently recognized pestivirus species. Phylogenetic analysis of the complete coding sequences of these two strains, along with 13 other sequences representing the four established species, indicated that CSFV, BDV, and Reindeer-1 have bifurcated from one common branch and BVDV-1 and BVDV-2 from another. In the former branch BDV and the pestivirus from reindeer are more similar to each other than to CSFV. The giraffe pestivirus is equally distinct from both major branches. In addition, the antigenic relatedness of pestivirus isolates covering the observed major genetic groups was studied by cross-neutralization assays. A clustering procedure on the basis of antigenic differences indicated the presence of six major groups corresponding to the genetically defined groups. Taken together, the results of our analyses addressing both nucleotide sequence relatedness and serological relatedness argue for the inclusion of Giraffe-1 and Reindeer-1 as the first members of two separate novel species within the genus Pestivirus. PMID- 11485414 TI - Translational activation of encapsidated potato virus X RNA by coat protein phosphorylation. AB - Previously we showed that encapsidated potato virus X (PVX) RNA is nontranslatable in vitro, but can be converted into a translatable form after binding to PVX particles of PVX-coded movement protein, the product of the first gene of triple gene block (TGBp1). Here we report that a similar effect occurs via in situ phosphorylation of the PVX coat protein (CP) by Ser/Thr protein kinase (PK) C, the mixture of casein kinases I and II or by cytoplasmic PK(s) from Nicotiana glutinosa leaves. Immunochemical analyses indicated that phosphorylation induced conformational changes in PVX CP. The N-terminal region of the PVX CP, rich in Ser and Thr residues, is exposed at the virion surface and can be removed by treatment with trypsin. We showed that (i) trypsin treatment removed the bulk of (32)P-radioactivity from in situ phosphorylated PVX CP, (ii) PVX containing N-terminally truncated CP (PVX-Ptd) failed to be translationally activated by phosphorylation, and (iii) the specific infectivity of PVX-Ptd was reduced. However, the PVX-Ptd RNA remained intact and PVX-Ptd could be translationally activated by the PVX MP TGBp1. We hypothesize that phosphorylation of the parental PVX by cytoplasmic PK(s) in vivo renders PVX RNA translatable in primary inoculated cells, whereas translational activation of the progeny virions destined for plasmodesmata trafficking is triggered by TGBp1. PMID- 11485415 TI - Frequency of direct repeat deletion in a human immunodeficiency virus type 1 vector during reverse transcription in human cells. AB - Retroviral genetic rearrangements can result from reverse transcriptase template switching. Most published data suggest that errors such as base misincorporation occur at similar frequencies for HIV-1 and for simple retroviruses such as spleen necrosis virus (SNV) and murine leukemia virus (MuLV). However, previous reports have suggested that template switch-mediated recombination is much more frequent for HIV-1 than for simple retroviruses. In this report, direct repeat deletion vectors similar to those previously used for measuring template switching events for SNV and MuLV were developed for HIV-1. Forward mutation rates and the frequency of template switching during a single cycle of HIV-1 replication were determined. The frequency of HIV-1-mediated repeat deletion was measured for three separate internal repeats in lacZ and was compared to rates observed with identical repeats for MuLV. The results indicated that the error rate and the frequency of repeat deletion of HIV-1 were similar to those of MuLV. PMID- 11485416 TI - High efficiencies of gene transfer with immobilized recombinant retrovirus: kinetics and optimization. AB - We used a combination of mathematical modeling and experiments to investigate the rate-limiting steps of retroviral transduction on surface-bound fibronectin (FN) and identify the conditions that maximize the efficiency of gene transfer. Our results show that fibronectin-assisted gene transfer (FAGT) is a strong function of the time and temperature of virus incubation in FN-coated plates. Gene transfer increases sharply at short times, reaches a maximum at intermediate times, and eventually declines as a result of loss of retroviral activity. The maximum transduction efficiency and the time at which this is attained increase with decreasing temperature of virus incubation. Depending on the temperature and the type of target cells, the initial rate of gene transfer increases by 3- to 10 fold and the maximum transduction efficiency increases by 2- to 4-fold as compared to traditional transduction (TT). Interestingly, Polybrene (PB) inhibits FAGT in a dose-dependent manner by inhibiting binding of retrovirus to FN. In contrast to traditional transduction, FAGT yields higher than 10-fold transduction efficiencies with concentrated retrovirus stocks. Gene transfer is directly proportional to the concentration of the virus-containing medium with no sign of saturation for the range of concentrations tested. These results suggest that immobilization of recombinant retrovirus can be rationally optimized to yield high efficiency of gene transfer to primary cells and improve the prospect of gene therapy for the treatment of human disease. PMID- 11485417 TI - Classification of static behavior of a class of unstructured models of continuous bioprocesses. AB - The stability characteristics of a class of unstructured models of continuous bioreactors are analyzed using elementary concepts of singularity theory and continuation techniques. The class consists of models for which the non-biomass product formation rate is linearly proportional to the utilization rate of limiting substrate. The kinetics expressions of cell growth and product synthesis are allowed to assume general forms of substrate and product. Global analytical conditions are derived that allow the construction of a practical picture in the multidimensional parameter space delineating the different static behavior these models can predict, including unique steady states, coexistence of non-trivial steady states with wash-out conditions, and multistability resulting from hysteresis. These general results are applied to specific examples of bioprocesses and allow the study of the effect of kinetic and operating parameters on the stability characteristics of these models. PMID- 11485418 TI - Effects of in situ cobalt ion addition on the activity of a gfp-oph fusion protein: the fermentation kinetics. AB - The effects of cobalt ion addition and inducer concentration were studied in the fermentation of E. coli BL21 expressing a GFP (green fluorescent protein)-OPH (organophosphorus hydrolase) fusion protein. It was found that cobalt ion addition improved the OPH activity significantly. When 2 mM of CoCl(2) was supplied during the IPTG-induction phase, OPH activity was enhanced approximately 10-fold compared to the case without cobalt or by the addition of cobalt to the cell extracts. Results indicate, therefore, that incorporation of the cobalt during synthesis is needed for enhanced activity. Also, the maximum OPH activity was not linearly related to inducer concentration. A mathematical model was then constructed to simulate these phenomena. Model parameters were determined by constrained least-squares and optimal IPTG and cobalt addition concentrations were obtained, pinpointing the conditions for the maximum productivity. Finally, the GFP fluorescence intensity was found linear to the OPH activity in each fermentation, demonstrating the function of GFP for monitoring its fusion partner's quantity in the bioreactor. PMID- 11485419 TI - Enhancing the atom economy of polyketide biosynthetic processes through metabolic engineering. AB - Polyketides, a large family of bioactive natural products, are synthesized from building blocks derived from alpha-carboxylated Coenzyme A thioesters such as malonyl-CoA and (2S)-methylmalonyl-CoA. The productivity of polyketide fermentation processes in natural and heterologous hosts is frequently limited by the availability of these precursors in vivo. We describe a metabolic engineering strategy to enhance both the yield and volumetric productivity of polyketide biosynthesis. The genes matB and matC from Rhizobium trifolii encode a malonyl CoA synthetase and a putative dicarboxylate transport protein, respectively. These proteins can directly convert exogenous malonate and methylmalonate into their corresponding CoA thioesters with an ATP requirement of 2 mol per mol of acyl-CoA produced. Heterologous expression of matBC in a recombinant strain of Streptomyces coelicolor that produces the macrolactone 6-deoxyerythronolide B results in a 300% enhancement of macrolactone titers. The unusual efficiency of the bioconversion is illustrated by the fact that approximately one-third of the methylmalonate units added to the fermentation medium are converted into macrolactones. The direct conversion of inexpensive feedstocks such as malonate and methylmalonate into polyketides represents the most carbon- and energy efficient route to these high value natural products and has implications for cost-effective fermentation of numerous commercial and development-stage small molecules. PMID- 11485420 TI - Factors affecting the production of a single-chain antibody fragment by Aspergillus awamori in a stirred tank reactor. AB - A recombinant strain of Aspergillus awamori expressing anti-lysozyme single chain antibody fragments (scFv), under the control of a xylanase promoter, was studied in order to investigate the impact of medium, induction regime and protease production on the expression of the product. Experiments with the time of induction showed that the optimum results are achieved when induction is started in the late exponential phase (21 h after inoculation) improving the titer of the product from 14.5 mg L(-1), obtained in the early exponential phase (7 h after inoculation), to 16.2 mg L(-1). A 100% increase of the carbon (fructose) and nitrogen (ammonium sulfate) sources in the growth medium resulted in an increase in product concentration from 16.2 to 108.9 mg L(-1) and an increase in maximum dry cell weight from 7.5 to 11.5 g L(-1). A 50% reduction in the concentration of the inducer resulted in an increase in the product yield from 10 mg g(-1) dry cell weight to 12 mg g(-1). Proteolytic enzymes were produced during the fermentation up to concentrations equivalent to 1.4 g L(-1) trypsin, but they had no detrimental effect on the concentration of the antibody fragment. PMID- 11485421 TI - Cell growth and by-product formation in a pyruvate kinase mutant of E. coli. AB - In this paper, we report on the analysis of acid formation in an E. coli pyk mutant. The results demonstrate that acid formation is insignificant for both the wild-type and the mutant at low glucose concentrations. However, at relatively high glucose concentrations, acid formation remains very low for the mutant but is significant for the wild-type. This substantial reduction in acids is accompanied by an increase in CO(2) production. Moreover, unlike the B. subtilis pyk mutant, the E. coli pyk mutant did not show a substantial increase in the PEP pool. PMID- 11485422 TI - Scale-up of a high cell density continuous culture with Pichia pastoris X-33 for the constitutive expression of rh-chitinase. AB - The feasibility of large-scale production of recombinant human chitinase using a constitutive Pichia pastoris expression system was demonstrated in a 21-L continuous stirred tank reactor. A steady-state recombinant protein concentration of 250 mg/L in the supernatant was sustained for 1 month at a dilution rate of 0.042 h(-1) (equivalent to one volume exchange per day), enabling a volumetric productivity of 144 mg/L d (240 U/L d). The steady-state dry cell weight concentration in this high cell density culture reached 110 g/L. Considering safety and economical aspects, all large-scale cultivations were conducted without molecular oxygen supplementation. Conventional air sparging was used instead. The oxygen demand of the process was determined by off-gas analysis (OUR = 4.8 g O(2) L(-1) h(-1) with k(L)a = 846 h(-1)) and evaluated with regard to further reactor scale-up. PMID- 11485423 TI - Identification and control of dissolved oxygen in hybridoma cell culture in a shear sensitive environment. AB - The productivity of mammalian cells can be enhanced by facilitating adequate oxygen transfer into the cultivation medium. However, current methods of controlling dissolved oxygen (DO) fail to account for alterations in medium composition during the course of the fermentation. These changes, which directly affect gas solubility and overall mass transfer coefficient, may be significant and deteriorate controller's performance in the long run. In this paper, the applications of Generalized Predictive Controllers (GPC) to DO control were investigated in a shear sensitive environment and compared to PID and Model Predictive Controllers (MPC). Input and output data for system identification were initially generated by varying the composition of oxygen fed into the bioreactor from 0 to 0.21 mol % while keeping the total inlet gas flow rate at 8.75 vvm. The process was identified using an AutoRegressive model with eXogeneous inputs (ARX) model and tested on different data sets. The model parameters were then correlated with the overall mass transfer coefficients. In simulation tests, the output of the PID controller switched from minimum to maximum values while more continuous control signals were obtained with the MPC and GPC controllers. When tested in a cell-free medium, all three controllers were able to track setpoint changes with some chattering observed in the control signals. The GPC outperformed the MPC and PID controllers when applied to the cultivation of hybridoma cells. PMID- 11485424 TI - Caseinolytic activity of fruit extract from Opuntia ficus-indica on bovine, caprine, and ovine sodium caseinates. AB - The rates and extents of hydrolysis of alpha(S)- and beta-caseins from bovine, caprine, and ovine sodium caseinates produced by an enzymatic extract of the fruit of Opuntia ficus-indica, (L.) Miller were evaluated and compared with those produced by a commercial animal rennet. A mechanistic model based on a pseudo first-order enzymatic reaction, in the presence of first-order deactivation of the enzyme, was postulated and successfully fitted to the experimental data. The animal rennet exhibited higher enzymatic efficiency than the fruit extract, irrespective of the source (i.e., bovine, caprine, or ovine) and the type (i.e., alpha(S)- or beta-casein) of substrate. The enzymatic efficiency (k(cat)/K(m)) for alpha(S)-casein ranged from 72 to 220 and from 43 to 65 L g(-1) h(-1), and for beta-casein from 242 to 742 and from 55 to 164 L g(-1) h(-1), for the animal rennet and the enzymatic extract of O. ficus-indica, respectively. Finally, it was observed that beta-casein from caprine and ovine caseinates was degraded by O. ficus-indica faster than its alpha(S) counterpart, but the reverse was observed for bovine caseinate. PMID- 11485425 TI - Bifurcation analysis of continuous biochemical reactor models. AB - The validity of a biochemical reactor model often is evaluated by comparing transient responses to experimental data. Dynamic simulation can be a rather inefficient and ineffective tool for analyzing bioreactor models that exhibit complex nonlinear behavior. Bifurcation analysis is a powerful tool for obtaining a more efficient and complete characterization of the model behavior. To illustrate the power of bifurcation analysis, the steady-state and transient behavior of three continuous bioreactor models consisting of a small number of ordinary differential equations are investigated. Several important features, as well as potential limitations, that are difficult to ascertain via dynamic simulation are disclosed through the bifurcation analysis. The results motivate the use of dynamic simulation and bifurcation analysis as complementary tools for analyzing the nonlinear behavior of bioreactor models. PMID- 11485426 TI - Hairy roots of Brugmansia candida that grow without agitation: biotechnological implications. AB - Hairy roots of Brugmansia candida that grew without agitation were obtained. Kinetics of growth and production of the tropane alkaloids scopolamine and hyoscyamine, with and without agitation, were studied. The exponential growth rate was higher in the roots that were exposed to shaking (0.13 d(-1)) than in the nonagitated ones (0.09 d(-1)). The specific production and the levels per flask of both alkaloids were enhanced without shaking. The use of these roots in large-scale productions could be economically advantageous. It remains to be seen if the data obtained in shake flasks can be extrapolated to large-scale bioreactors. PMID- 11485427 TI - Adsorption step in the biological degradation of a textile dye. AB - This research documents the removal of the dye Gris Lanaset G from aqueous solutions by fungal pellets. Adsorption of the dye by dead biomass pellets of Trametes versicolor was determined and compared with dye removal by enzymatic degradation. Six kinetic equations were fitted to the experimental adsorption data obtained. The results indicate that kinetics such as the Elovich equation, which considers that the rate-controlling step is the diffusion of the dye molecules, show the best fit. Nonlinear Langmuir and Freundlich equations were also fitted into the adsorption data, and it can be concluded that the adsorption equilibrium can be interpreted by the Langmuir isotherm. Adsorption plays an important role in the process of the elimination of color from textile wastewater, although not all of the elimination is due to this physical process when the microorganism is active. The removal of color (around 90%) with active microorganisms is greater than that obtained with the adsorption process. PMID- 11485428 TI - Study and mathematical modeling of the production of propionic acid by Propionibacterium acidipropionici immobilized in a stirred tank fermentor. AB - A mathematical model was developed that describes production of propionic acid by fermentation of sweet whey with Propionibacterium acidipropionici immobilized in calcium polygalacturonate beads in a fermentor-type stirred tank. This mathematical model is constituted by a partial differential equations system, which fits consumption, production, growth and internal diffusion rates in the support. Fermentation was experimentally studied with free cells and immobilized cells, effective diffusivities of lactose and propionic acid were estimated in the support, and typical parameters of the model were obtained by nonlinear regression of the experimental data. The variance analysis shows that the combination of micro(max) and K(d) parameters is the source of variation most significative, also they were found to be the most sensitive parameters of the model. Finally, an effectiveness factor was calculated in order to assess the effect of mass transfer on the overall reaction rate observed. PMID- 11485430 TI - Optimal synthesis of protein purification processes. AB - There has been an increasing interest in the development of systematic methods for the synthesis of purification steps for biotechnological products, which are often the most difficult and costly stages in a biochemical process. Chromatographic processes are extensively used in the purification of multicomponent biotechnological systems. One of the main challenges in the synthesis of purification processes is the appropriate selection and sequencing of chromatographic steps that are capable of producing the desired product at an acceptable cost and quality. This paper describes mathematical models and solution strategies based on mixed integer linear programming (MILP) for the synthesis of multistep purification processes. First, an optimization model is proposed that uses physicochemical data on a protein mixture, which contains the desired product, to select a sequence of operations with the minimum number of steps from a set of candidate chromatographic techniques that must achieve a specified purity level. Since several sequences that have the minimum number of steps may satisfy the purity level, it is possible to obtain the one that maximizes final purity. Then, a second model that may use the total number of steps obtained in the first model generates a solution with the maximum purity of the product. Whenever the sequence does not affect the final purity or more generally does not impact the objective function, alternative models that are of smaller size are developed for the optimal selection of steps. The models are tested in several examples, containing up to 13 contaminants and a set of 22 candidate high-resolution steps, generating sequences of six operations, and are compared to the current synthesis approaches. PMID- 11485429 TI - Comparison of growth and recombinant protein expression in two different insect cell lines in attached and suspension culture. AB - Culture conditions required for obtaining maximum recombinant protein concentrations from two cell lines, Spodoptera frugiperda (IPLBeta-Sf21-AE) and Trichoplusia ni (Tn 5Beta-1-4), were determined in this work. Conditions studied include mode of culture (suspended vs attached), agitation rates, inoculum sizes, cell concentration at the time of infection, and various serum-free media (SFM). Results were compared with the performance of attached cultures in TnM-FH with 10% fetal bovine serum. Growth rates in the different culture media tested were similar, but the cell numbers achieved (i.e., yield) improved 2 to 2.7-fold in SFM over cultures in TnM-FH. Agitation rates of 150-160 rpm were necessary for maximum growth of suspended Tn 5Beta-1-4 cells compared to 125-150 rpm for Sf-21 cells. An inoculum size of 5 x 10(5) cells/mL gave good growth rates and optimum biomass yields for both cell lines. Cultures of both cell lines were infected with viruses encoding for beta-galactosidase or human secreted alkaline phosphatase (seAP). Protein expression in TnM-FH in attached culture showed that Tn 5Beta-1-4 cells are 2-4.5 times more productive on a per cell basis than Sf-21 cells grown under similar conditions. Production of beta-galactosidase in Sf-21 cells increased 50% in suspension cultures with SFM compared to attached cultures in TnM-FH, but seAP expression was essentially unchanged by culture techniques. The Tn 5Beta-1-4 cells produced 2.6-4.4 and 2.7-3 times more beta-galactosidase and seAP, respectively, in SFM in suspension compared to Sf-21 cells. EX-CELL 401 and Sf900-II were formulated as optimized SFM for Sf cell lines. However, in Sf 21 cultures EX-CELL 400 performed better than the other two media, as it increased the beta-galactosidase yield up to 25%. Surprisingly, EX-CELL 401 was the best medium for the production of beta-galactosidase by Tn 5Beta-1-4 cells, resulting in 25% and 69% higher volumetric and specific yields, respectively, compared to EX-CELL 405 which was formulated for this specific cell line. These results show that even when culture media are designed for maximal growth of a specific cell line, other media may provide the best conditions for protein production. PMID- 11485431 TI - Assembly and mixing of two-stage aqueous two-phase protein extractions. AB - The effect of the order of system assembly was studied in two-stage aqueous two phase systems loaded with whole bovine blood and BSA. Recovery in the bottom phase of the back extraction varied by up to 40% depending on the manner in which the forward extraction was assembled, significantly impacting upon the efficiency of the process. The effect of mixing intensity was investigated using a simple shear device. As expected, higher shear rates were found to facilitate mass transfer and shorten the time taken to reach equilibrium. Mixing at lower shear rates for extended periods resulted in a lower recovery of protein. PMID- 11485432 TI - Water activity, pH and density of aqueous amino acids solutions. AB - The water activity, pH and density of some aqueous amino acid solutions were determined at 25 degrees C in three different types of solvents. Previous published experimental data on water activity and solubility of amino acids in aqueous solutions were used together with data from this work to test the applicability of a group contribution model. The activity coefficients were estimated by the UNIFAC-Larsen model combined with the Debye-Huckel equation, taking also into account the partial dissociation phenomena of species in solution. Interaction energies between the charged species Na(+) and Cl(-) and the specific groups of amino acids (COOH and NH(2)) were adjusted using experimental solubility data. PMID- 11485433 TI - Comparison of energy-minimized structures of [Pd(II)(N-methyliminodiacetate)] complexes of X(1)-His-X(3)-His-His peptides as an analysis of steric and specific interactions with synthetic binding tags for IMAC separations. AB - [Pd(II)(mida)(peptide)] complexes for the series of peptides of sequence X(1)-His X(3)-His-His were studied by molecular mechanics methods using Spartan, MMFF94, and SYBYL programs with X(1) = X(3) = glycine (G), phenylalanine (F), tyrosine (Y), tryptophan (W), and with X(1) = glycine (G) and X(3) = proline (P). For comparison purposes, data were also obtained for the Ser-Pro-His-His-Gly (SPHHG) and the (His)(5) peptides.The latter two peptides and GHPHH are tags in current use for IMAC separations. These provide calibration points as to the binding affinities that have been determined for the entire series. The energies of the complexes, as an average trend found from the composite behavior of the three methods, were found to be SPHHG (205 kcal/mol) (most stable; are values obtained by MMFF94 methods) < HH(#)HH(#)H(#) (222; where # implies the site of attachment to match the other X(1)-His-X(3)-His-His peptides) < YHYHH (249) < GHGHH (265) < WHWHH (284) approximately GHPHH (286) < FHFHH (311) (least stable), implying that FHFHH might be a useful chromatographic tag for IMAC protein separations that would elute more readily than GHPHH from IMAC sites that are of square-planar structure, such as Cu(II)(ida-supported) IMAC columns. Specific H-bonded interactions are observed between the tyrosine X(1) and pendant carboxylates and between X(3) and the N-terminal amine of [Pd(mida)(YHYHH)]. Face-to-pi-face ring stacking occurs between phenylalanine X(1) and X(3) units in [Pd(mida)(FHFHH)], whereas edge C-H to pi H-bonding or pi stacking occurs between the X(1) and X(3) tryptophans of [Pd(mida)(WHWHH)]. Two energy minima were found with tryptophan. The more stable form has the aromatic rings more parallel, similar to the stacked form of phenylalanine, rather than the edge C-H to pi H-bonding, and virtually the same overall energy as for [Pd(mida)(GHPHH)]. The "perpendicular" structure was found as an initial local energy minimum, but additional MMFF94 calculations found the pi -stacked arrangement at energy ca. 39 kcal/mol lower than that of the nearly "perpendicular" arrangement of the tryptophan rings, a composite effect of relaxation of the peptide, together with differences in stabilities imparted by the differing geometries. The use of the terms "pi-stacked " and "perpendicular" forms represent the limiting cases available to the tryptophan side chain groups. A twist of about 15 degrees to 20 degrees in dihedral angle is all that is necessary to change between structures that are nearly described as one form or the other. PMID- 11485434 TI - Protein purification by fusion with an environmentally responsive elastin-like polypeptide: effect of polypeptide length on the purification of thioredoxin. AB - Elastin-like polypeptides (ELPs) undergo a reversible, soluble-to-insoluble phase transition in aqueous solution upon heating through a characteristic transition temperature (T(t)). Incorporating a terminal ELP expression tag into the gene of a protein of interest allows ELP fusion proteins to be purified from cell lysate by cycles of environmentally triggered aggregation, separation from solution by centrifugation, and resolubilization in buffer. In this study, we examine the effect of ELP length on the expression and purification of a thioredoxin-ELP fusion protein and show that reducing the size of the ELP tag from 36 to 9 kDa increases the expression yield of thioredoxin by 4-fold, to a level comparable to that of free thioredoxin expressed without an ELP tag, while still allowing efficient purification. However, truncation of the ELP tag also results in a more complex transition behavior than is observed with larger tags. For both the 36 kDa and the 9 kDa ELP tag fused to thioredoxin, dynamic light scattering showed that large aggregates with hydrodynamic radii of approximately 2 microm form as the temperature is raised to above the T(t). These aggregates persist at all temperatures above the T(t) for the thioredoxin fusion with the 36 kDa ELP tag. With the 9 kDa tag, however, smaller particles with hydrodynamic radii of approximately 12 nm begin to form at the expense of the larger, micron-size aggregates as the temperature is further raised above the T(t). Because only large aggregates can be effectively retrieved by centrifugation, efficient purification of fusion proteins with short ELP tags requires selection of solution conditions that favor the formation of the micron-size aggregates. Despite this additional complexity, our results show that the ELP tag can be successfully truncated to enhance the yield of a target protein without compromising its purification. PMID- 11485435 TI - Selectivity of IMAC columns in trypsin inhibitor purification. AB - The properties of an adsorbent and the parameters in an adsorption process affect the resolution of chromatographic purifications. This is reflected in the elution profile, which shows the relative affinity of different proteins for a specific adsorbent. In the work presented here, elution profiles for trypsin inhibitor were used to study the effects of the concentration of trypsin inhibitor, ionic strength of the protein solution, slope of the elution gradient, and the regeneration treatment of the chromatography column on the selectivity of the adsorbent Cellufine Chelate-Cu(II)(ida). Cytochrome c was used as a reference protein. Variations in the concentrations of trypsin inhibitor and in the ionic strength of the buffered solution did not have any effects on the elution profile. On the other hand, changes in the slope of the pH gradient used for elution caused shifting of the elution peaks toward lower values of the elution volume, resulting in the best strategy to modify the elution profile of the system. Finally, using a constant slope pH gradient of elution, the variation of the selectivity of the adsorbent for trypsin inhibitor when subjected to cleaning treatments with 0.5 N NaOH was studied. Appropriate cleaning practices used in industry were followed. The adsorbent showed only a slight tendency for resolution loss in the order of 2 x 10(-4) days(-1). The results presented here show a good stability of the adsorbent when compared to other biospecific adsorbents commonly used. PMID- 11485436 TI - Lipase immobilization into porous chitoxan beads: activities in aqueous and organic media and lipase localization. AB - Lipases were noncovalently immobilized in Chitoxan, a polyionic hydrogel obtained by complexation between chitosan and xanthan. The properties of free and immobilized lipases have been compared. In the aqueous medium, the activity was twice as high for immobilized lipases as for free lipases. Immobilized lipases in chitoxan were able to hydrolyze triacylglycerols in three distinct organic solvent media. At the microstructural level, lipases were not distributed uniformly in the chitoxan beads. Higher concentrations of lipase were found in the outer membrane-like layer of the beads, as compared with lower concentrations in the inner part of the beads. PMID- 11485437 TI - Agar-based magnetic affinity support for protein adsorption. AB - Magnetic colloidal particles were prepared by a coprecipitation method. The particles were composed of nanometer-sized superparamagnetic Fe(3)O(4) particles stabilized by lauric acid. Then, magnetic agar gel beads were produced by a water in-oil emulsification method using a mixture of agar solution and the magnetic colloidal particles as the aqueous phase. A reactive triazine dye, Cibacron blue 3GA (CB), was coupled to the gel to prepare an agar-based magnetic affinity support (MAS) for protein adsorption. The support showed good magnetic responsiveness in a magnetic field. Bovine serum albumin (BSA) was used as a model protein to test adsorption equilibrium and kinetic behavior of the MAS. The adsorption equilibrium of BSA to the MAS was described by the Langmuir-type isotherm. Adsorption capacity of the MAS for BSA was up to 25 mg/mL at a CB coupling density of 1.6 micromol/mL. The effect of ionic strength on BSA adsorption was complex, exhibiting a maximum capacity at an ionic strength of 0.06 mol/L. The adsorption of BSA to the MAS was also influenced by pH. Uptake rate of BSA to the MAS was analyzed using a pore diffusion model. The pore diffusion coefficient was estimated to be 1.75 x 10(-11) m(2)/s. Finally, recycled use of the MAS demonstrated the stability of the MAS in protein adsorption and magnetic responsiveness. PMID- 11485438 TI - Pressure-flow relationships for packed beds of compressible chromatography media at laboratory and production scale. AB - Pressure drop across chromatography beds employing soft or semirigid media can be a significant problem in the operation of large-scale preparative chromatography columns. The shape or aspect ratio (length/diameter) of a packed bed has a significant effect on column pressure drop due to wall effects, which can result in unexpectedly high pressures in manufacturing. Two types of agarose-based media were packed in chromatography columns at various column aspect ratios, during which pressure drop, bed height, and flow rate were carefully monitored. Compression of the packed beds with increasing flow velocities was observed. An empirical model was developed to correlate pressure drop with the aspect ratio of the packed beds and the superficial velocity. Modeling employed the Blake-Kozeny equation in which empirical relationships were used to predict bed porosity as a function of aspect ratio and flow velocity. Model predictions were in good agreement with observed pressure drops of industrial scale chromatography columns. A protocol was developed to predict compression in industrial chromatography applications by a few laboratory experiments. The protocol is shown to be useful in the development of chromatographic methods and sizing of preparative columns. PMID- 11485439 TI - Hyperaccumulation of nickel by hairy roots of alyssum species: comparison with whole regenerated plants. AB - Hairy roots were used to investigate nickel uptake by the hyperaccumulator species, Alyssum bertolonii, A. tenium, and A. troodii. The Ni biosorption capacity of A. tenium hairy roots was lower than for other types of biomass such as bacteria and algae; in short-term (9-h) equilibrium studies, the highest Ni content measured in the roots was 17 500 microg g(-1) dry weight at a liquid concentration of about 4000 ppm. Using long-term hairy root cultures, it was demonstrated that Ni tolerance and hyperaccumulation do not necessarily depend on the presence of shoots or root-shoot translocation. A. bertolonii hairy roots remained healthy in appearance and continued to grow in the presence of 20-100 ppm Ni, accumulating up to 7200 microg g(-1) dry weight Ni. In contrast, hairy roots of Nicotiana tabacum turned dark brown at 20 ppm Ni and growth was negligible. The ability to grow at high external Ni concentrations allowed hyperaccumulator hairy roots to remove much greater amounts of heavy metals from the culture liquid than nonhyperaccumulator hairy roots, even though biomass Ni concentrations were similar. Although hairy roots proved to be a useful tool for investigating Ni hyperaccumulation, there were significant differences in the Ni uptake capacity of hairy roots and whole plants. Regenerated plants of A. tenium were much more tolerant of Ni and capable of accumulating higher Ni concentrations than hairy roots of this species. PMID- 11485440 TI - Role of viscoelastic properties of differentiated SH-SY5Y human neuroblastoma cells in cyclic shear stress injury. AB - Shear stress and strain lead to neurodegeneration in vivo during head injury, glaucoma, and certain repetitive motion disorders. In vitro, shear stress and strain have been shown to lead to cell injury in a number of models using neurons and neuron-like cells. In the present study we examined the relationship between shear stress, strain, and the extent of cell injury in a cyclic shear stress induced model of cell injury using differentiated SH-SY5Y (human neuroblastoma) cells. Shear stress led to cell strain that increased with increasing stress and diminished upon cessation of shear. Strain rate during cyclic application of shear stress increased by over an order of magnitude from the first to all subsequent cycles, suggesting that the cell and/or its polymer network became more elastic upon cyclic shear stress application. To support this conclusion we measured the degree of cytoskeletal polymerization before and after exposure of cells to cyclic shear stress and found that the fraction of polymerized tubulin in the cell relative to total tubulin decreased by a factor of 2 after six cycles of shear stress. The extent of injury, as indicated by the fraction of cells with fragmented DNA, was three times higher for cyclic shear stress than for steady shear stress and may be related to the change in strain rate and/or cytoskeletal reorganization associated with cyclic stress. These findings may aid in understanding the mechanism by which neurons and neuron-like cells respond to cyclic shear stress and strain and lead to new treatments for disease or injury arising from the exposure of neurons to abnormal cyclic or repetitive stress and strain. PMID- 11485441 TI - Analysis of a microbial community oxidizing inorganic sulfide and mercaptans. AB - Successful treatment of refinery spent-sulfidic caustic (which results from the addition of sodium hydroxide solutions to petroleum refinery waste streams) was achieved in a bioreactor containing an enrichment culture immobilized in organic polymer beads with embedded powdered activated carbon (Bio-Sep). The aerobic enrichment culture had previously been selected using a gas mixture of hydrogen sulfide and methyl mercaptan (MeSH) as the sole carbon and energy sources. The starting cultures for the enrichment consisted of several different Thiobacilli spp. (T. thioparus, T. denitrificans, T. thiooxidans, and T. neopolitanus), as well as activated sludge from a refinery aerobic wastewater treatment system and sludge from an industrial anaerobic digester. Microscopic examination (light and SEM) of the beads and of microbial growth on the walls of the bioreactor revealed a great diversity of microorganisms. Further characterization was undertaken starting with culturable aerobic heterotrophic microorganisms (sequencing of PCR amplified DNA coding for 16S rRNA, Gram staining) and by PCR amplification of DNA coding for 16S rRNA extracted directly from the cell mass, followed by the separation of the PCR products by DGGE (denaturing gradient gel electrophoresis). Eight prominent bands from the DGGE gel were sequenced and found to be closest to sequences of uncultured Cytophagales (3 bands), Gram-positive cocci (Micrococcineae), alpha proteobacteria (3 bands), and an unidentified beta proteobacterium. Culturable microbes included several genera of fungi as well as various Gram-positive and Gram-negative heterotrophic bacteria not seen in techniques using direct DNA extraction. PMID- 11485442 TI - Calorimetric studies on dry pectinlyase preparations: impact of glass transition on inactivation kinetics. AB - The glass transition temperature (T(g)) of a dry ultrafiltrated pectinlyase (PL) preparation decreased from 56 to 24 degrees C when water content increased to 20%. The thermal transition temperature (T(p)) for protein denaturation decreased greatly up to 40% moisture; above 40% no further changes in T(p) were observed. In the glassy state, a lag period of approximately 7 days with no PL activity loss was observed; after that, PL activity was lost. Above T(g), the rates of PL inactivation greatly increased. In the glassy state E(a) was 16.6 kJ/mol. When the system was in a higher mobility state (rubbery), E(a) increased to 66.5 kJ/mol. PMID- 11485443 TI - A droplet medium selection for cultivation of Bacillus thuringiensis. AB - For an industrial fermentation system, design of a suitable medium for cultivation of cells is an important step. Medium selection is commonly carried out with a lot of experimental works. In the present study, a simple method was proposed for medium selection in cultivation of Bacillus thuringiensis for production of thuringiensin. A droplet of medium was employed to test the suitability of the medium. PMID- 11485444 TI - Effect of addition of water-soluble polysaccharides on bacterial cellulose production in a 50-L airlift reactor. AB - Bacterial cellulose (BC) production was carried out in a batch cultivation of Acetobacter xylinum in a 50-L internal loop airlift reactor by addition of water soluble polysaccharides into the medium. When 0.1% (w/w) agar was added, BC production reached 8.7 g/L compared with 6.3 g/L in the control, and duration of the cultivation period to reach the maximum concentration of BC was almost half of that without addition of polysaccharides. During cultivation, BC was formed into pellets whose size was smaller when the productivity of BC was higher, indicating that increase in the relative viscosity by addition of polysaccharides hindered formation of large clumps of BC and increase in the volumetric oxygen transfer coefficient at high flow rate led to increase in BC productivity. PMID- 11485445 TI - Physicochemical characteristics of LR3-IGF1 protein inclusion bodies: electrophoretic mobility studies. AB - A knowledge of the physicochemical properties of inclusion bodies is important for the rational design of potential recovery processes such as flotation and precipitation. In this study, measurement of the size and electrophoretic mobility of protein inclusion bodies and cell debris was undertaken. SDS-PAGE analysis of protein inclusion bodies subjected to different cleaning regimes suggested that electrophoretic mobility provides a qualitative measure of protein inclusion body purity. Electrophoretic mobility as a function of electrolyte type and ionic strength was investigated. The presence of divalent ions produced a stronger effect on electrophoretic mobility compared with monovalent ions. The isoelectric point of cell debris was significantly lower than that for the inclusion bodies. Hence, the contaminating cell debris may be separated from inclusion bodies using flotation by exploiting this difference in isoelectric points. Separation by this method is simple, convenient, and a possible alternative to the conventional route of centrifugation. PMID- 11485446 TI - The cis-trans isomerization of 1,2,5,6-tetrasilacycloocta-3,7-dienes: analysis by mechanistic probes and density functional theory. AB - A series of alkyl- and aryl-substituted derivatives of cis,cis-1,2,5,6 tetrasilacycloocta-3,7-diene were prepared. Isomerization of these compounds to the corresponding trans,trans-1,2,5,6-tetrasilacycloocta-3,7-dienes by exposure to Ru and Zr hydride complexes was explored. Experimental probes of the isomerization were consistent with a stepwise mechanism involving metal hydride addition/elimination rather than one involving radical intermediates. Analysis of the low energy conformers of the various cis and trans isomers of 1,1,2,2,5,5,6,6 octamethyl-1,2,5,6-tetrasilacycloocta-3,7-diene using density functional theory suggested the following trend in stability: trans,trans > cis,trans > cis,cis. The calculated trend in stability was consistent with the experimentally observed unidirectional isomerization of the carbon-carbon double bonds from all cis to all trans and supports a cis,trans isomer as a tenable intermediate. PMID- 11485447 TI - Kinetics of the cis,cis to trans,trans isomerization of 1,1,2,2,5,5,6,6 octamethyl-1,2,5,6-tetrasilacycloocta-3,7-diene. AB - The kinetics of the ruthenium-promoted cis,cis to trans,trans isomerization of 1,1,2,2,5,5,6,6-octamethyl-1,2,5,6-tetrasilacycloocta-3,7-diene were investigated. Incubation of a ruthenium alkylidene complex, (Cy(3)P)RuCl(2)(==CHPh)Ru(p-cymene)Cl(2), in CD(2)Cl(2) for 5 days at 40 degrees C afforded a catalytically active ruthenium species that was shown to be responsible for promoting the isomerization. The isomerization was observed to proceed in two steps: (1) conversion of the starting cis,cis isomer to a proposed cis,trans intermediate and (2) subsequent conversion of the intermediate to the product trans,trans isomer. Kinetic studies demonstrated that the two steps are first-order with respect to the concentrations of the cis,cis isomer, the intermediate, and the ruthenium alkylidene complex. The data were further consistent with a mechanism involving bimolecular hydride addition-elimination during the two isomerization steps. PMID- 11485448 TI - Ring-closing metathesis of olefinic peptides: design, synthesis, and structural characterization of macrocyclic helical peptides. AB - Heptapeptides containing residues with terminal olefin-derivatized side chains (3 and 4) have been treated with ruthenium alkylidene 1 and undergone facile ring closing olefin metathesis (RCM) to give 21- and 23-membered macrocyclic peptides (5 and 6). The primary structures of peptides 3 and 4 were based upon a previously studied heptapeptide (2), which was shown to adopt a predominantly 3(10)-helical conformation in CDCl(3) solution and an alpha-helical conformation in the solid state. Circular dichroism, IR, and solution-phase (1)H NMR studies strongly suggested that acyclic precursors 3 and 4 and the fully saturated macrocyclic products 7 and 8 also adopted helical conformations in apolar organic solvents. Single-crystal X-ray diffraction of cyclic peptide 8 showed it to exist as a right-handed 3(10)-helix up to the fifth residue. Solution-phase NMR structures of both acyclic peptide 4 and cyclic peptide 8 in CD(2)Cl(2) indicated that the acyclic diene assumes a loosely 3(10)-helical conformation, which is considerably rigidified upon macrocyclization. The relative ease of introducing carbon-carbon bonds into peptide secondary structures by RCM and the predicted metabolic stability of these bonds renders olefin metathesis an exceptional methodology for the synthesis of rigidified peptide architectures. PMID- 11485449 TI - Syntheses of 5a'-homo-vinblastine and congeners designed to establish structural determinants for isolation of atropisomers. AB - The syntheses of 5a'-homo-vinblastine (3a) and its C-20' methyl congener 62a were achieved. In contrast to vinblastine, these compounds did not allow isolation of atropisomers because of their lower conformational inversion barrier. However, annelation of a six-membered ring to the conformationally mobile D'-piperidine ring provided an isolated atropisomer 81a, which could be converted to its lower energy conformation 65a on heating. The 5a'-homo-vinblastine congeners 3a, 62a, and 65a showed vinblastine-like inhibition of tubulin polymerization and cytotoxicity to L1210 leukemia cells, albeit at lower potency for the latter activity, than that found with the corresponding compounds in the vinblastine series. PMID- 11485450 TI - The syntheses of 16a'-homo-leurosidine and 16a'-homo-vinblastine. Generation of atropisomers. AB - The synthesis of 16a'-homo-leurosidine was achieved through enantioselective generation of a ring D'-seco-precursor 33 (without requirement of a chiral auxiliary). Its cyclization provided the N(b')-quaternary salt 35 with a configuration corresponding to the atropisomeric form 8a rather than 8b of the target product. On debenzylation, the amine 8a was obtained and found not to isomerize thermally to the anticipated atropisomer 8b (in contrast to its lower homologue, with its formation of natural leurosidine). However, on protonation, a 1:1 mixture of atropisomers of 16a'-homo-leurosidine was obtained. A synthesis of 16a'-homo-vinblastine provided two atropisomers 5a and 5b for the free base at equilibrium (1:2.3 at room temperature in CDCl(3)), with a shift to the major conformer 5b with increasing solvent acidity or decreasing temperature. The synthesis was achieved through a stereoselective inversion of the tertiary hydroxyl function in the enantioselectively generated C-20' progenitor 39. PMID- 11485452 TI - Carbanion-induced base-catalyzed synthesis of 1H-isothiochromenes, benzo[c]thiochromenes through ring-transformation reactions of 6-aryl-2H-pyran-2 ones. AB - An innovative approach to the one-pot synthesis of highly functionalized 3,4 dihydro-1H-isothiochromenes (3), 6H-benzo[c]thiochromenes (5, 6), 6H benzo[c]chromenes (8), and 2,3-dihydro-1-benzothiophenes (10, 11) is delineated from the reaction of a suitably functionalized 6-aryl-3-carbomethoxy-4-methylthio 2H-pyran-2-one (1) and a carbanion generated from tetrahydrothiopyran-4-one, 4 thiochromanone, 4-chromanone, and tetrahydrothiophene-3-one through ring transformation reactions. PMID- 11485451 TI - Novel annulene dications from methylated [2.2]metacyclophane monoenes and [e] ring benzannelated dimethyldihydropyrene. AB - Tetramethyl- and hexamethyl-substituted [2.2]metacyclophane monoenes (10 and 11) are transformed into their corresponding trans dimethyldihydroethanophenanthrenium dications (14(2+) and 15(2+)) in FSO(3)H x SbF(5) (4:1) and FSO(3)H x SbF(5) (1:1) with SO(2)ClF or SO(2) as the solvent; these 10 pi-dications are equivalent to the C-4/C-5 diprotonated dications of the 2,7-dimethyl derivative of trans-DMDHP, 3a. The trans-12c,12d-dimethyl-12c,12d dihydrobenzo[e]pyrene (6) reacts with FSO(3)H/SO(2)ClF under surprisingly mild conditions to give initially a persistent diprotonated dication (6H(2)(2+)) and, subsequently, the oxidation dication (6(2+)); the 6(2+):6H(2)(2+) ratio reaches 4:1 after 1 week at low temperature. Protonation of the anti-metacyclophane (13) was also examined. Charge delocalization mode and tropicity in the resulting dications are gauged via detailed NMR studies at 500 MHz. PMID- 11485454 TI - Calorimetric and computational study of 1,3,5-trithiane. AB - To understand the differences in conformational behavior and reactivity of oxygen and sulfur-containing 1,3,5-heterocyclohexanes, the enthalpies of formation and sublimation of 1,3,5-trithiane, 1, have been measured. The numerical value of the enthalpy of formation for this compound in the solid state is -8.6 +/- 2.6 kJ mol(-1), while the corresponding value in the gaseous state is 84.6 +/- 2.6 kJ mol(-1). The value for the enthalpy of sublimation is 93.2 +/- 0.2 kJ mol(-1). Standard ab initio molecular orbital calculations at the G2(MP2), G2, and G3 levels were performed, and the calculated enthalpies of formation are compared with the experimental data. These experimental and theoretical studies support the relevance of through-space lone pair-lone pair electronic repulsion in the sulfur heterocycle. PMID- 11485453 TI - Role of singlet diradicals in reactions of 2-carbenabicyclo[3.2.1]octa-3,6-diene. AB - The generation of 2-carbenabicyclo[3.2.1]octa-3,6-diene (1) results in the formation of C(8)H(8) hydrocarbons endo-6-ethynylbicyclo[3.1.0]hex-2-ene (4), semibullvalene (5), and 5-ethynyl-1,3-cyclohexadiene (6), and C(8)H(10) hydrocarbons bicyclo[3.2.1]octa-2,6-diene (7), tricyclo[3.2.1.0(4,6)]oct-2-ene (8), and tetracyclo[3.3.0.0(2,8)0(4,6)]octane (9). Focus is placed on three mechanistic pathways for the formation of the C(8)H(10) hydrocarbon fraction: (a) abstraction of hydrogen by triplet carbene 1T to produce an equilibrating set of monoradicals, (b) interconversion of triplet carbene 1T into tricyclic triplet diradical 19T and tetracyclic triplet diradical 20T, and (c) interconversion of singlet 1S with analogous singlet diradical 19S and 20S. Ab initio calculations at the (U)B3LYP/6-311+G(3df,2p)//(U)B3LYP/6-31G(d,p) and broken spin symmetry UBS B3LYP/6-311+G(3df,2p)//B3LYP/6-31G(d,p) levels rule out choices (a) and (b) and are consistent with the singlet diradical process. PMID- 11485455 TI - Diels--Alder bioconjugation of diene-modified oligonucleotides. AB - In an effort to offer complementary technology for covalent biomolecule modification (bioconjugation), we have developed a method that exploits the aqueous acceleration of Diels--Alder reactions for this purpose. Three different diene phosphoramidite reagents have been synthesized that enable diene modification of synthetic oligonucleotides prepared by the phosphoramidite method. Clean and efficient Diels--Alder cycloaddition of these diene oligonucleotides with maleimide dieneophiles was carried out, and the labeled oligonucleotide bioconjugates were characterized by HPLC and electrospray mass spectrometry. Dieneophile stoichiometry, temperature, and pH are all parameters that were shown to influence the efficiency of the process. PMID- 11485456 TI - Hydroboration. 97. Synthesis of new exceptional chloroborane--Lewis base adducts for hydroboration. Dioxane--monochloroborane as a superior reagent for the selective hydroboration of terminal alkenes. AB - Several less volatile oxygen-containing Lewis bases, such as tert-butyl methyl ether, dioxane, anisole, ethyl acetate, beta-chloroethyl ether, and monoglyme, were examined as prospective mono- and dichloroborane carriers. Dioxane, ethyl acetate, and beta-chloroethyl ether form relatively stable boron trichloride adducts, but the boron trichloride adduct of monoglyme is not very stable and must be used immediately. On the other hand, tert-butyl methyl ether and anisole fail to form stable boron trichloride adducts and the corresponding ether-cleaved products are obtained. Among the selected oxygen-containing Lewis bases, only dioxane forms stable and reactive mono- and dichloroborane adducts. Monoglyme and beta-chloroethyl ether give stable dichloroborane adducts requiring excess of diborane. Convenient methods for the preparation of mono- and dichloroborane adducts of dioxane from dioxane-BCl(3) and NaBH(4) in the presence of catalytic amounts of tri- or tetraglyme were developed. The dioxane--monochloroborane adduct hydroborates representative olefins cleanly and rapidly. The corresponding alcohols were obtained in quantitative yields after oxidation. Also, the hydroboration of several terminal olefins with dioxane--monochloroborane were highly regioselective and the primary alcohols were obtained almost exclusively (>99.5%), after oxidation. Accordingly, dioxane-monochloroborane should serve as a reagent of choice for such hydroborations. The dioxane--dichloroborane adduct showed remarkable selectivity toward 2-substituted terminal olefins, such as 2 methyl-1-butene and beta-pinene, when compared to simple terminal and hindered olefins, giving a unique tool for selective hydroborations. Dichloroborane adducts of monoglyme and beta-chloroethyl ether also showed high reactivity, even at room temperature, toward simple unhindered olefins. However, hydroboration of hindered olefins is slow and requires either higher temperatures or the addition of 1 equiv of boron trichloride to liberate free dichloroborane, as in the case of the previously known dichloroborane adducts of methyl sulfide and diethyl ether. PMID- 11485457 TI - Synthesis of several halobisnoradamantane derivatives and their reactivity through the S(RN)1 mechanism. AB - Several bridgehead halobisnoradamantane derivatives (5, 7, 10, and 17) were synthesized from tricyclic diester 1 in good yields using standard methods. The reactivity through the S(RN)1 mechanism of the above compounds and the known halobisethano derivatives 24 and 25a-c was studied. Iodo derivatives 7, 10, and 25a reacted with diphenylphosphide ions in DMSO under irradiation to give the corresponding substitution and reduction products by the S(RN)1 mechanism, while iodo ketone 17 gave a mixture of the rearranged substitution product 36 and the reduction product 18. Formation of 36 takes place through a 1,5-hydrogen migration of the initially formed radical, a kind of process that has been observed for the first time in the S(RN)1 propagation steps. The diiodo derivative 24 reacted with diphenylphosphide ions under similar reaction conditions to give the substitution and/or reduction products 32, 31, 27, 25a, and 26. The intramolecular ET reaction in the monosubstitution radical anion 32*( ) seems to be faster than the intermolecular ET to the substrate, and the monoiodo derivative 25a is a reaction intermediate. PMID- 11485458 TI - New methodologies for the preparation of porphodimethenes and their conversion to trans-porphyrins with functionalized naphthyl spacers. AB - The MacDonald [2 + 2]-type condensation of readily available 5-aryl-substituted dipyrromethanes with acenaphthenequinone leads to the trans-syn- and anti porphodimethenes, which in turn can be converted to the alpha,alpha- and alpha,beta-porphyrin atropisomers, respectively. Treatment of the metalated or unmetalated porphodimethenes with KOH or NaOMe in THF followed by protonation with HCl results in a ring opening of the acenaphthenone and formation of the trans-8-carboxynaphthylporphyrins or their esters (NaOMe) after oxidation. Alternatively, the porphyrin formation can be accomplished by reaction of the porphodimethenes with acids in the presence of water or methanol. Reaction with NaBH(4) in a THF--methanol mixture yields the corresponding dialcohols in nearly quantitative yields. Sixteen different building blocks were prepared in order to evaluate the generality of this new synthetic approach, with Ar = 2,4,6 Me(3)C(6)H(2); 2,6-Cl(2)C(6)H(3); 2,6-F(2)C(6)H(3); 3,4-tBu(2)C(6)H(3); 3,4,5 (MeO)(3)C(6)H(2); 4-BrC(6)H(4); 4-MeC(6)H(4); and 4-MeOOCC(6)H(4) at the meso positions. The synthesized porphodimethenes and porphyrins have been fully characterized, and the X-ray structure analyses of three representative derivatives are presented. PMID- 11485459 TI - Resolution of an iridoid synthon, gastrolactol, by means of dynamic acetylation and lipase-catalyzed alcoholysis. AB - A short synthetic route to asymmetric iridoids was developed. The three key steps were an intramolecular [4 + 2] cycloaddition reaction of an enamine derivative of 8-oxocitral (2), a dynamic acetylation, and an enzymatic resolution of the gastrolactyl acetates 5a and 5b, iridoids with three stereocenters. Some regio- and stereoselective heterogeneous catalytic hydrogenations of double bonds in iridoid aglucones were discussed. PMID- 11485460 TI - Stereospecific alkyl and alkynyl substitution reactions of epoxy sulfides with organoaluminums with double inversion of the configuration. AB - A regioselective and stereospecific substitution reaction of 1-(phenylthio)-2,3 epoxyalkanes was achieved by using organoaluminum reagents as a nucleophile. Under the influence of trimethyl- or triethylaluminum, a 1-(phenylthio)-2,3 epoxyalkane underwent substitution at the C2 position to give a product with retention of the configuration. The reaction proceeds through an episulfonium ion intermediate, which gives rise to the C2-substitution products with double inversion of the configuration. Introduction of an alkynyl group was also accomplished by the reaction with dimethyl[2-(trimethylsilyl)ethynyl]aluminum in dichloromethane. PMID- 11485461 TI - Reaction of vinylcarbenoids with benzaldehydes: formation of vinylcarbonyl ylides followed by ring closure to oxiranes and dihydrofurans. AB - Rh(2)(OAc)(4)-catalyzed reaction of vinyldiazo compound 1a in the presence of p methoxybenz- 2a, mesit- 2b, and p-chlorobenzaldehyde 2c gave a mixture of isomeric vinyloxiranes 3a-c and 4a-c, and sterically unstable (E)-dihydrofurans 5a-c, but not stable (Z)-dihydrofurans 6. However, the reactions with p-nitro- 2d and 2,4-dinitrobenzaldehyde 2e gave (Z)-dihydrofurans 6d,e along with 3d, 4d, and 5d,e. The reaction in the presence of maleic anhydride and dimethyl fumarate gave single 1,3-dipolar cycloadducts 11 and 13, respectively, indicating that a single conformer, the sterically unstable endo-aryl-endo-cyanostyryl carbonyl ylide 14 (15), is initially formed in the reaction of 1 with 2. It was concluded that the endo-vinyl-exo-aryl vinylcarbonyl ylides 17a-c arising from 2a-c undergo disrotatory cylization to exclusively produce 5, whereas the ylides 17d,e arising from 2d,e undergo partly symmetry-forbidden conrotaory cyclization to sterically stable trans-diaryldihydrofurans 6d,e as well as the symmetry-allowed process to 5d,e. PMID- 11485462 TI - Water-soluble adamantane-terminated dendrimers possessing a rhenium core. AB - A novel type of radiometal-containing dendrimer with potential radiotherapeutical applications is described. Different generations of this adamantane-terminated, Frechet-type dendrimer (28, 29, 30), each consisting of two dendritic wedge ligands around a rhenium core, have been synthesized in organic solvent via reaction with ReO(PPh(3))(2)Cl(3). Through complexation of their adamantane groups by beta-cyclodextrins (beta-CDs), these dendrimers were made water soluble (9.6, 0.4, and 0.2 mM, respectively). beta-CD-induced solubilization of the wedges in water allowed the complexes to be made under aqueous conditions, via reaction with rhenium gluconate. Not only does this strategy enable the facile synthesis of the radioactive analogue, the yields for these complex-formation reactions in water also turned out to be far higher than those observed for the reactions in organic solvents. PMID- 11485463 TI - Efficient Baylis--Hillman reaction using stoichiometric base catalyst and an aqueous medium. AB - A practical and efficient set of conditions were developed using stoichiometric base catalyst, 1,4-diazabicyclo[2,2,2]octane (DABCO), and an aqueous medium to overcome problems commonly associated with the Baylis--Hillman reaction, such as low reaction yields and long reaction time. These simple modifications to the classical conditions, using more base catalyst and an aqueous medium, proved to be successful in converting a variety of aliphatic and aromatic aldehydes to their corresponding Baylis--Hillman products. The inclusion of environmentally friendly water in the reaction solvent was critical for achieving the high yield of Baylis--Hillman adducts. Our deuterium-exchange experiments suggest that the Michael addition adduct formed between DABCO and methyl acrylate is the active intermediate for the Baylis--Hillman reaction in aqueous conditions, and its hydrolysis, a nonproductive side reaction facilitated by the quaternary ammonium ion, leading to the formation of a stable betaine product, consumes both the catalyst and methyl acrylate, making it necessary to add more base catalyst and methyl acrylate. PMID- 11485464 TI - Synthesis of fluorescent diarylethenes having a 2,4,5-triphenylimidazole chromophore. AB - Diarylethenes 1a-4a, having a fluorescent 2,4,5-triphenylimidazole chromophore in the aryl group, were synthesized. Upon excitation of the triphenylimidazole chromophore with 366 nm, 1a-4a underwent photocyclization reactions, and the solutions containing 1a-4a changed color from colorless to red-purple or to blue. The colors disappeared by irradiation with visible (lambda > 480 nm) light. The fluorescence intensity of the solutions also reversibly changed with the photochromic reactions. The fluorescence quantum yields of 1a, 2a, 3a, and 4a were determined to be 4.6, 7.7, 9.1, and 8.4%, respectively. The fluorescence quantum yields decreased with the increase in photocyclization quantum yields. PMID- 11485465 TI - Synthesis of beta-lactones by the regioselective, cobalt and Lewis acid catalyzed carbonylation of simple and functionalized epoxides. AB - The PPNCo(CO)(4) and BF(3) x Et(2)O catalyzed carbonylation of simple and functionalized epoxides in DME gives the corresponding beta-lactones regioselectively in good to high yields. The carbonylation occurred selectively at the unsubstituted C-O bond of the epoxide ring, and this reaction tolerates various functional groups such as alkenyl, halide, hydroxy, and alkyl ether. PMID- 11485466 TI - Cyclofunctionalization and free-radical-based hydrogen-transfer reactions. An iterative reaction sequence applied to the synthesis of the C(7)-C(16) subunit of zincophorin. AB - The strategy considered herein features an iodocyclofunctionalization/hydrogen transfer reaction sequence for the elaboration of propionate motifs. Proceeding with excellent yield and diastereoselectivity, the synthetic sequence proposed gives access to the anti-anti dipropionate motif when the reduction step is performed under the control of the exocyclic effect. The tandem sequence is applied successfully to the synthesis of the C(7)-C(16) subunit of zincophorin, and iteration of the process gives the desired anti-anti-anti-anti polypropionate stereopentad. Modifications of the reaction sequence--including phenylselenocyclofunctionalization, carbonate hydrolysis, and chelation controlled radical reduction reactions--lead to the formation of the anti-syn dipropionate motif with remarkable diastereocontrol. PMID- 11485467 TI - Enantiocontrolled preparation of indolizidines: synthesis of (-)-2 epilentiginosine and (+)-lentiginosine. AB - A highly stereoselective approach to (-)-2-epilentiginosine and (+)-lentiginosine has been developed based on a diastereofacially selective cycloaddition of dichloroketene with a chiral dienol ether. The two naturally occurring indolizidines are each obtained enantioselectively (> or = 99:1) in ca. 8.5% overall yield. PMID- 11485468 TI - Theoretical prediction of a base-catalyzed bicyclic Boulton--Katritzky rearrangement. AB - On the basis of quantum chemical calculations, the bicyclic Boulton--Katritzky rearrangement (BKR) has been classified as a pseudopericyclic reaction. Theoretical investigations extend the applicability of the BKR to classes of molecules other than those known from experiments. PMID- 11485469 TI - Synthetic approaches to a variety of covalently linked porphyrin--fullerene hybrids. AB - There is substantial interest in dyads in which C(60) is covalently linked to electron donors, such as porphyrins, which absorb light strongly in the visible region. We present here the details of the syntheses of such compounds, which can be broadly organized into categories depending upon the nature of the linker joining the two chromophores. The structural aspects of intramolecular electronic interaction that we have sought to explore have dictated the synthetic strategies employed to generate these classes of molecules. Flexible glycol linkers were used to allow close approach between the fullerene and porphyrin, facilitating through-space interactions. These linkers also allowed studies of the effects of metal cation complexation. Naphthalene and alkyne linkers were used to examine the possible effects a conjugated or aromatic linker might have on photophysical properties. Finally, steroids were used as linkers in dyads expected to possess a large distance between the two chromophores, in which only through-bond interactions between the fullerene and porphyrin should be possible. PMID- 11485470 TI - Thermochemical and kinetic studies of a bisphenol antioxidant. AB - The results of a thermodynamic and kinetic investigation on the homolytic reactivity of 3,3'-di-tert-butyl-5,5'-dimethyl(1,1'-biphenyl)-2,2'-diol (1) are reported. EPR studies of the equilibration between 1, 2,4,6-trimethylphenol, and the corresponding radicals obtained by abstraction of a hydroxylic hydrogen allowed us to determine the OH bond dissociation energy (BDE) of investigated bisphenol as 83.10 kcal/mol. This value is considerably larger than that reported for the structurally related 2,6-di-tert-butyl-4-methylphenol (BHT), i.e., 81.02 kcal/mol. Absolute rate constants for the reaction of 1 with alkyl, alkoxyl, and peroxyl radicals, at or nearly to room temperature, were also determined by competition kinetics in the first two cases and by autoxidation studies under controlled conditions in the last one. The experimental data indicate that this bisphenol is a moderately efficient antioxidant and polymerization inhibitor. PMID- 11485471 TI - ((15)N(5))-labeled adenine derivatives: synthesis and studies of tautomerism by (15)N NMR spectroscopy and theoretical calculations. AB - Since the nitrogens of nucleosides and nucleotides play an important role in the molecular recognition of these compounds, (15)N NMR became a method of choice in this field. Fully (15)N-labeled adenine, required in the latter studies, was obtained in four synthetic steps, in a good yield. Likewise, ((15)N(5))-2 hexylthioether-adenine and ((15)N(5))-8-Br-adenine were obtained in five synthetic steps from the relatively inexpensive (15)N sources: (15)N-NH(4)Cl, (15)N-NH(4)OH, (15)N-NaNO(2). Full (15)N labeling of these adenine prototypes enabled to obtain high-resolution (15)N NMR spectra of these bases at 60.8 MHz. Furthermore, the spectra suggested the existence of the N3-H species in the tautomeric mixtures of these compounds in solution, in addition to the well reported N9-H (major) and N7-H (minor) tautomers. These observations were also supported by quantum mechanical calculations of the tautomeric equilibria in the gas phase and in solution of the above-mentioned adenine compounds. The gas-phase tautomeric equilibria were estimated using density functional theory and second order perturbation theory methods. Solvent effects were included by means of both continuum and discrete solvation models. The observation of the existence of the N3-H tautomer has a clear impact on the possible H-bonding patterns of these adenine prototypes and on their molecular recognition by various biological macromolecules. The above(15)N-labeled analogues are expected to find use as (15)N NMR probes for numerous biochemical studies. PMID- 11485472 TI - Intramolecular AR--O--AR bond formation in calixarenes. AB - The formal dehydration of two vicinal phenol moieties of p-tert butylcalix[6]arene was achieved in two steps by mild oxidation of the calixarene followed by treatment of the resulting monospirodienone derivative (9c) with an ionic hydrogenation mixture (Et(3)SiH/CF(3)COOH). Reaction of 9c yielded the unsubstituted xanthenocalix[6]arene 11d, while treatment of the monospirodienone derivative of a spherand-type calixarene (13) with Et(3)SiH/CF(3)COOH afforded the dibenzofuran derivative 15. The formation of the latter product indicates that, at least for 13, the rings forming the Ar--O--Ar bond in the product are not those connected by the spiro bond in the starting material. Methylation of the phenolic hydroxyl groups of 11d with methyl p-toluenesulfonate/K(2)CO(3) or dimethyl sulfate/base afforded its dimethyl and tetramethyl ether derivatives. The parent xanthone calix[6]arene derivative 17b was prepared by O-methylation of the phenol groups followed by CrO(3) oxidation of the xanthene methylene group and deprotection of the OH groups. McMurry coupling of calixanthone 17a afforded the dixanthylene 18. Calixarenes 11d and 15 (which possess a xanthene and dibenzofuran group, respectively) were structurally characterized by X-ray crystallography. PMID- 11485473 TI - A stereoselective approach for the synthesis of alpha-sialosides. AB - A highly efficient synthesis of the human melanoma associated antigen GD(3) derivative has been described. A key feature of the synthetic approach was the use of sialyl donors that were protected with a C-5 trifluoroacetamide moiety. These sialyl donors gave high yields and excellent alpha-anomeric selectivities in direct glycosylations with a wide variety of glycosyl acceptors ranging from C 8 hydroxyls of sialic acids and C-3 hydroxyls of galactosides to reactive primary alcohols. PMID- 11485474 TI - Novel bicyclic nucleoside analogue (1S,5S,6S)-6- hydroxy-5-hydroxymethyl-1 (uracil-1-yl)-3,8-dioxabicyclo[3.2.1]octane: synthesis and incorporation into oligodeoxynucleotides. AB - The novel bicyclic nucleoside (1S,5S,6S)-6-hydroxy-5-hydroxymethyl-1-(uracil-1 yl)-3,8-dioxabicyclo[3.2.1]octane [2'-deoxy-1'-C,4'-C-(2-oxapropano)uridine] (15), expected to be restricted into an O4'-endo furanose conformation, was synthesized from the known 1-(3'-deoxy-beta-D-psicofuranosyl)uracil 5. The phosphoramidite derivative of 15 was successfully incorporated into oligodeoxynucleotides using standard methods, and thermal denaturation studies showed moderate decreases in duplex stabilities of -2.1 and -1.5 degrees C per modification toward complementary DNA and RNA, respectively. PMID- 11485475 TI - Synthesis of natural flutimide and analogous fully substituted pyrazine-2,6 diones, endonuclease inhibitors of influenza virus. AB - Flutimide, a fully substituted 1-hydroxy-3H-pyrazine-2,6-dione, is a fungal metabolite isolated from a new species of Delitschia cofertaspora. It has been shown to selectively inhibit cap-dependent endonuclease activity of influenza virus A. The inhibition of this activity is a target for the potential development of a therapeutic agent to treat influenza infections. A convergent total synthesis of flutimide starting from L-leucine has been described. The synthetic methodology has been extended to include the synthesis of specifically designed aromatic analogues of flutimide, some of which exhibited greater than 7 fold improvement in activity. The most potent compounds were those with p fluorobenzylidene or p-methoxybenzylidene substitutions at C-5 of 3H-pyrazine-2,6 dione and showed IC(50) values of 0.9 and 0.8 microM, respectively. The details of the rationale for the synthetic design, syntheses, and biological activities of these analogues are described. PMID- 11485476 TI - Polyacene and cyclacene geometries and electronic structures: bond equalization, vanishing band gaps, and triplet ground states contrast with polyacetylene. AB - The ground-state geometries and excited singlet and lowest triplet energies of polyacenes from benzene through nonacene are predicted with B3LYP/6-31G* calculations and compared to experimental data where available. The results are compared to these data for cyclacenes and polyenes. The polyacenes and cyclacenes have geometries consisting of two fully delocalized nonalternating ribbons joined by relatively long bonds. Polyacenes are predicted to have smaller band gaps than the corresponding polyenes and triplet ground states for nine or more benzene rings. The fully delocalized nonalternating nature of polyacenes differs from the bond alternation resulting from Peierls distortion in polyenes. The differences are rationalized in terms of a simple MO model, and the results are compared to extensive prior theoretical work in the literature. Predictions about the electronic structure of analogues containing polyacene units are made. PMID- 11485477 TI - Fluorescence-based screening of asymmetric acylation catalysts through parallel enantiomer analysis. Identification of a catalyst for tertiary alcohol resolution. AB - A technique for high-throughput screening of kinetic resolution catalysts is reported. The method relies on carrying simultaneous kinetic resolutions in a multiwell plate format wherein each well contains a unique catalyst and a small amount of a pH-activated fluorescent sensor (3). By conducting experiments such that each catalyst is evaluated in parallel in the presence of each isolated enantiomer, an indication of catalyst activity is obtained on a per enantiomer basis. Catalysts that are highly active for one enantiomer but modestly active for another are then reevaluated in conventional kinetic resolutions. From these screens, a highly selective (k(rel) = 46) pentapeptide (4) was obtained for a model secondary alcohol (1). In addition, peptide 10 was found to afford excellent selectivities (k(rel) > 20) for a number of alcohol substrates (9a-9f) in the traditionally challenging tertiary class. PMID- 11485478 TI - Vinyl sulfones in solid-phase synthesis: preparation of 4,5,6,7 tetrahydroisoindole derivatives. AB - The preparation of functionalized 4,5,6,7-tetrahydroisoindole via a traceless solid-phase sulfone linker strategy is described. Thermolytic extrusion of SO(2) from polymer-bound 3-(phenylsulfonyl)-3-sulfolene (7) generated polymer-bound 2 (phenylsulfonyl)-1,3-butadiene (9) in situ which underwent Diels--Alder cycloaddition with various dienophiles to furnish vinyl sulfone resins 10-14. To complete a traceless linker cleavage strategy, (p-tolysulfonyl)methyl isocyanide or ethyl isocyanoacetate was employed to react with the vinyl sulfone moiety to liberate functionalized 4,5,6,7-tetrahydroisoindole products from the resin. Using this chemistry, nine tetrahydroisoindole derivatives (6, 15-22) were prepared in 32-41% overall yields from polystyrene/divinylbenzene sulfinate 1. PMID- 11485479 TI - Nitrogen kinetic isotope effects on the decarboxylation of 4-pyridylacetic acid. AB - Nitrogen kinetic isotope effects on the decarboxylation of 4-pyridylacetic acid have been measured in solvents of different polarity and have been found to vary from the inverse value of 0.994 to the normal value of 1.002 upon increase of water content of the binary dioxane--water solvent from 25% to 75% (v/v), respectively. These changes were successfully modeled theoretically and shown to originate from the large inverse nitrogen isotope effect on the equilibrium between acidic and zwitterionic forms. PMID- 11485480 TI - Analysis of amide bond formation with an alpha-hydroxy-beta-amino acid derivative, 3-amino-2-hydroxy-4-phenylbutanoic acid, as an acyl component: byproduction of homobislactone. AB - In the synthesis of peptidomimetics containing alpha-hydroxy-beta-amino acid, the coupling of this N(beta)-protected beta-amino acid with amine components was generally performed without the protection of its alpha-hydroxyl group. However, the formation of dipeptides in low yield was often observed when sterically hindered amine components were used. Boc-Apns-OH [Apns: (2S,3S)-3-amino-2-hydroxy 4-phenylbutanoic acid, allophenylnorstatine] (6), which is one of such beta-amino acid derivatives, is intensively employed as a core structure in the development of HIV-1 protease inhibitors. There have been no precise studies, to date, that have examined amide bond formation with alpha-hydroxy-beta-amino acid derivatives as an acyl component. To determine the cause of this low-yield reaction, we studied the amide bond formation focusing on the activation step of N(beta) protected alpha-hydroxy-beta-amino acid by using a model coupling reaction between 6 and H-Dmt-OR [Dmt: (R)-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid] (7). A significant amount of homobislactone 9 was formed through the activation of the carboxyl group of 6 to the benzotriazole-type active esters such as OBt and OAt. In addition, this homobislactone formation was markedly increased in the presence of a catalytic amount of a base, which exhibited good correlation with the low yield of the amide bond formation, suggesting that homobislactone formation is one major reason for the low yield of the amide bond formation. Moreover, homobislactones were also formed in other derivatives of the N(beta) protected alpha-hydroxy-beta-amino acid, suggesting a common feature of this type of amino acids. The use of a strong activation method like EDC--HOAt without base addition enhanced amide bond formation, although a small amount of homobislactone may be formed during the coupling reaction. PMID- 11485481 TI - An asymmetric approach to spirocylic systems: a formal synthesis of zizaene. AB - A general route to enantiopure spirocarbocycles is described. The use of various chiral bicyclic lactams 1 that have been doubly alkylated with olefinic halides gives good yields of alpha,alpha-disubstituted chiral lactams 2 which were cyclized to spiro-olefins using ring closure metathesis methodology (Grubbs' catalyst). These spirolactams 3, formed in generally excellent yields, were shown to be smoothly transformed into spirocyclopentenone 6, spirocyclohexenone, 7, and spirolactams 8. Further demonstration of this spirocyclization methodology was featured in a formal synthesis of zizaene, by preparing in enantiomeric form the Coates' intermediate 21. This synthetic effort provided additional examples of the synthetic versatility of chiral bicyclic lactams 2a,b. PMID- 11485482 TI - Stereoselective reactions of acyclic allylic phosphates with organocopper reagents. AB - A series of acyclic allylic alcohols of general structure R(1)CH==CHCH(OH)R(2) were resolved by Sharpless kinetic resolution. The hydroxyl groups of these enantiomerically enriched alcohols were derivatized to diethyl phosphates, and the derivatives were reacted with organocopper reagents. Cleanest substitution reactions were observed with reagents R(3)(2)CuCNLi(2). With R(1) = Me and R(3) = n-Bu, the size of R(2) affected both the regioselectivity and stereoselectivity of the displacement. Larger R(2) groups gave higher regio- and stereoselectivities: with R(2) = 3-pentyl, >98% S(N)2' regioselectivity and >98% anti stereoselectivity were observed. Bn(2)CuCNLi(2) gave stereoselectivities comparable to those observed with n-Bu(2)CuCNLi(2) but t-Bu(2)CuCNLi(2) exhibited much lower diastereofacial preference. PMID- 11485483 TI - Vinyl carbocations: solution studies of alkenyl(aryl)iodonium triflate fragmentations. AB - Generation of vinyl cations is facile by fragmentation of alkenyl(aryl)iodonium trifluoromethanesulfonates. Kinetics and electronic effects were probed by (1)H NMR spectroscopy in CDCl(3). Products of fragmentation include six enol triflate isomers in addition to iodoarenes. The enol triflates arise from direct reaction of a triflate anion with the starting iodonium salts as well as triflate reaction with rearranged secondary cations derived from those salts. G2 calculations of the theoretical isodesmic hydride-transfer reaction between secondary vinyl cation 7 and primary vinyl cation 6 reveal that cation 6 is 17.8 kcal/mol higher in energy. Activation parameters for fragmentation of (Z)-2-ethyl-1-hexenyl(3,5 bis-trifluoromethylphenyl)iodonium triflate, 17e, were calculated using the Arrhenius equation: E(a) = 26.8 kcal/mol, Delta H(++) = 26.2 kcal/mol, and Delta S(++) = 11.9 cal/mol x K. Added triflate increases the rate of fragmentation slightly, and it is likely that for most beta,beta-dialkyl- substituted vinylic iodonium triflates enol triflate fragmentation products are derived from three competing mechanisms: (a) vinylic S(N)()2 substitution; (b) ligand coupling (LC); and (c) concerted aryliodonio departure and 1,2-alkyl shift leading to secondary rather than primary vinyl cations. PMID- 11485484 TI - Synthesis of alpha-substituted beta-amidophosphines by diastereoselective alkylation. A new access to chiral ligands for asymmetric catalysis. AB - Chiral beta-amidophosphine boranes 7a-f can be diastereoselectively alkylated, using O-protected amino-alcohols as chiral inducers, to furnish alpha-substituted beta-amidophosphine boranes 8a-f and 9-12 with up to 72% diastereoisomeric excess. Selective deprotection afforded optically pure carboxylic derivative 13 which is a key intermediate for the synthesis of various potential chiral ligands for asymmetric catalysis. PMID- 11485485 TI - Effect of para substituents on the rate of bond shift in arylcyclooctatetraenes. AB - The rate constants for bond shift (k(BS)) in phenylcyclooctatetraene (1b) and its p-nitro and p-methoxy analogues (1a and 1c, respectively) in THF-d(8) were determined by dynamic NMR spectrometry to be identical, but k(BS) is eight times greater at 280 K relative to 1b when the para substituent is cyclooctatetraenyldipotassium (2(2-)/2K(+)). These results are discussed in the context of (a) possible intrinsically small substituent effects (as determined by (13)C chemical shifts in the ground state (GS)) for 1a-c and (b) differences in steric interactions and resonance stabilization between the ground and BS transition state (TS). The latter factor was modeled by employing HF/3-21G(*) ab initio molecular orbital calculations of the GS and ring inversion TS. It is concluded that k(BS) is unchanged in 1a-c because the potentially greater pi interaction in the BS TS is counterbalanced by a greater degree of twist between the aryl and COT rings resulting from increased steric hindrance relative to the GS. However, pi interaction assumes a greater importance in the TS of 2(2-)/2K(+) owing to a decreased HOMO-LUMO energy gap compared to 1a-c, particularly when the counterions are solvated. This causes a decrease in the inter-ring twist angle and, together, these changes are responsible for the observed increase in k(BS) in 2(2-)/2K(+). The effect of substituents on a possible contribution of heavy atom tunneling to the reaction mechanism is also discussed. PMID- 11485486 TI - Convergent total synthesis of khafrefungin and its inhibitory activity of fungal sphingolipid syntheses. AB - A convergent total synthesis of khafrefungin, a novel inhibitor of fungal sphingolipid syntheses isolated from the fermentation culture MF6020, has been developed. Alkenylboronic acid 5 and alkenyliodide 6, key fragments for the total synthesis, were prepared from the corresponding achiral aldehydes using tin(II) catalyzed and Zr(IV)-catalyzed asymmetric aldol reactions, respectively. The Suzuki coupling reaction of these two fragments was successfully performed to give 17 in good yield. Through the total synthesis, epimerization of the C4 position having a rather highly acidic proton did not occur, indicating that khafrefungin was under strict conformational constraints to prevent the epimerization process. This characteristic stability of khafrefungin has also been discussed using semiempirical calculation and synthesis. Finally, khafrefungin derivatives have also been synthesized, and their antifungal activities have been measured to obtain information on the structure--activity relationships. PMID- 11485487 TI - Conversions by dimethyldioxirane of 1-alkylbenzotriazoles into their N-oxides and of 2-alkylbenzotriazoles into 2-alkyl-trans-4,5,6,7-diepoxy-4,5,6,7 tetrahydrobenzotriazoles. AB - Dimethyldioxirane converted 1-alkylbenzotriazoles 4 to the corresponding 3 alkylbenzotriazole 1-oxides 5 in good yields, but transformed 2 alkylbenzotriazoles 12 into 2-alkyl-trans-4,5,6,7-diepoxy-4,5,6,7 tetrahydrobenzotriazoles 13. PMID- 11485488 TI - Syntheses of 1,4-benzothiazepines and 1,4-benzoxazepines via cyclizations of 1-[2 arylthio(oxy)ethyl]-5-benzotriazolyl-2-pyrrolidinones and 3-benzotriazolyl-2-[2 arylthio(oxy)ethyl]-1-isoindolinones. AB - 1-[2-Arylthio(oxy)ethyl]-5-benzotriazolyl-2-pyrrolidinones 6a-e, 12 and 3 benzotriazolyl-2-[2-arylthio(oxy)ethyl]-1-isoindolinones 9a-f, 14 are readily available from reactions of benzotriazole (4), 2-(arylsulfanyl)ethylamines 3, or 2-phenoxyethylamine (11) with 2,5-dimethoxy-2,5-dihydrofuran (5) or 2 formylbenzoic acid (8). Lewis acid mediated cyclizations of 6 and 9 produced novel 1,4-benzothiazepines 7a-e and 10a-f, respectively. Cyclizations of 12 and 14 gave 1,4-benzoxazepines 13 and 15, respectively. PMID- 11485489 TI - Efficient syntheses of thiochromans via cationic cycloadditions. AB - alpha-(Benzotriazolyl)methyl thioethers 1a-e reacted with styrenes under Lewis acid catalysis to give novel polysubstituted thiochromans (3,4-dihydro-2H-1 benzothiopyrans) 3-14 and 16-20 in generally high yields. Most thiochromans were isolated as one diastereomer following recrystallization. The configuration and conformation of the products are predicted on the basis of their NMR data. A stepwise reaction, proceeding via a [4(+) + 2] cationic polar cycloaddition mechanism, is proposed. PMID- 11485490 TI - Dibenzo[b,h][1,4,7]thiadiazonines: examples of a novel ring system. AB - Acetanilides 1a-e react with 1,1' sulfinylbis(benzotriazole)/trimethylchlorosilane at 45-65 degrees C to form 1,2 di(benzotriazol-1-yl)-2-arylimino-1-ethanethiones 3a-e, while heating the same reagents at 110 degrees C results in dibenzo[b,h][1,4,7]thiadiazonines 5a,c,d, and 6. X-ray crystal structures are reported for three representative examples. PMID- 11485491 TI - One-carbon homologation of carboxylic acids via BtCH(2)TMS: a safe alternative to the Arndt--Eistert reaction. AB - Carboxylic acids are converted into the corresponding homologated acids or esters, using easily available 1-(trimethylsilylmethyl)benzotriazole (1) as a one carbon synthon. The effectiveness of the reaction has been investigated on six aryl and seven alkyl carboxylic acids. PMID- 11485492 TI - Novel syntheses of 2,3-disubstituted benzofurans. PMID- 11485493 TI - Bromination studies of the 2,3-dimethylnaphthazarin core allowing easy access to naphthazarin derivatives. PMID- 11485494 TI - A catalytic cycle for the asymmetric synthesis of epoxides using sulfur ylides. PMID- 11485495 TI - Selective reduction of terminal alkynes to alkenes by indium metal. PMID- 11485496 TI - A concise synthesis of rac-staurosporine glycon. PMID- 11485497 TI - An efficient route to either enantiomer of trans-2-aminocyclopentanecarboxylic acid. PMID- 11485498 TI - Electrolytic partial fluorination of organic compounds. 47. Highly regioselective anodic monofluorination of 2-thiadiazolyl, 2-oxadiazolyl, and 2-triazolyl sulfides. PMID- 11485499 TI - Synthesis of (+)- and (-)-statine via chiral sulfoxide chemistry. PMID- 11485500 TI - Intramolecular 4 + 3 cycloadditions. Theoretical and experimental evaluation of endo/exo preferences of a cyclopentenyl cation. PMID- 11485501 TI - A sequential application of kinetic resolution and polymer-supported scavenging for the isolation of chiral secondary alcohols. PMID- 11485502 TI - A prospective study of physical activity and cognitive decline in elderly women: women who walk. AB - BACKGROUND: Several studies have suggested that physical activity is positively associated with cognitive function in elderly persons. Evidence about this association has been limited by the cross-sectional design of most studies and by the frequent lack of adjustment for potential confounding variables. We determined whether physical activity is associated with cognitive decline in a prospective study of older women. METHODS: We studied 5925 predominantly white community-dwelling women (aged > or =65 years) who were recruited at 4 clinical centers and were without baseline cognitive impairment or physical limitations. We measured cognitive performance using a modified Mini-Mental State Examination at baseline and 6 to 8 years later. Physical activity was measured by self reported blocks (1 block approximately 160 m) walked per week and by total kilocalories (energy) expended per week in recreation, blocks walked, and stairs climbed. Cognitive decline was defined as a 3-point decline or greater on repeated modified Mini-Mental State Examination. RESULTS: Women with a greater physical activity level at baseline were less likely to experience cognitive decline during the 6 to 8 years of follow-up: cognitive decline occurred in 17%, 18%, 22%, and 24% of those in the highest, third, second, and lowest quartile of blocks walked per week (P< .001 for trend). Almost identical results were obtained by quartile of total kilocalories expended per week. After adjustment for age, educational level, comorbid conditions, smoking status, estrogen use, and functional limitation, women in the highest quartile remained less likely than women in the lowest quartile to develop cognitive decline (for blocks walked: odds ratio, 0.66 [95% confidence interval, 0.54-0.82]; for total kilocalories: odds ratio, 0.74 [95% confidence interval, 0.60-0.90]). CONCLUSIONS: Women with higher levels of baseline physical activity were less likely to develop cognitive decline. This association was not explained by differences in baseline function or health status. This finding supports the hypothesis that physical activity prevents cognitive decline in older community dwelling women. PMID- 11485503 TI - Risk of recurrent coronary events in relation to use and recent initiation of postmenopausal hormone therapy. AB - BACKGROUND: The finding from the Heart and Estrogen/Progestin Replacement Study (HERS) of increased coronary risk restricted to the first year after starting postmenopausal hormone therapy raises new questions about the role of hormone therapy in women with coronary heart disease. We assessed the risk of recurrent myocardial infarction or coronary heart disease death associated with the use and recent initiation of hormone therapy in women who survived a first myocardial infarction. METHODS: The setting for this population-based inception cohort study was Group Health Cooperative, a health maintenance organization. We studied 981 postmenopausal women who survived to hospital discharge after their first myocardial infarction between July 1, 1986, and December 31, 1996. We obtained information on hormone use from the Group Health Cooperative computerized pharmacy database and identified recurrent coronary events by medical record review. RESULTS: During median follow-up of 3.5 years, there were 186 recurrent coronary events. There was no difference in the risk of recurrent coronary events between current users of hormone therapy and other women (adjusted relative hazard [RH], 0.96; 95% confidence interval [CI], 0.62-1.50). Relative to the risk in women not currently using hormones, there was a suggestion of increased risk during the first 60 days after starting hormone therapy (RH, 2.16; 95% CI, 0.94 4.95) and reduced risk with current hormone use for longer than 1 year (RH, 0.76; 95% CI, 0.42-1.36). CONCLUSION: These results are consistent with the findings from the HERS, suggesting a transitory increase in coronary risk after starting hormone therapy in women with established coronary heart disease and a decreased risk thereafter. PMID- 11485504 TI - The impact of diabetes mellitus on mortality from all causes and coronary heart disease in women: 20 years of follow-up. AB - BACKGROUND: Few data are available on the long-term impact of type 2 diabetes mellitus on total mortality and fatal coronary heart disease (CHD) in women. METHODS: We examined prospectively the impact of type 2 diabetes and history of prior CHD on mortality from all causes and CHD among 121 046 women aged 30 to 55 years with type 2 diabetes in the Nurses' Health Study who were followed up for 20 years from 1976 to 1996. RESULTS: During 20 years of follow-up, we documented 8464 deaths from all causes, including 1239 fatal CHD events. Compared with women with no diabetes or CHD at baseline, age-adjusted relative risks (RRs) of overall mortality were 3.39 (95% confidence interval [CI], 3.08-3.73) for women with a history of diabetes and no CHD at baseline, 3.00 (95% CI, 2.50-3.60) for women with a history of CHD and no diabetes at baseline, and 6.84 (95% CI, 4.71-9.95) for women with both conditions at baseline. The corresponding age-adjusted RRs of fatal CHD across these 4 groups were 1.0, 8.70, 10.6, and 25.8, respectively. Multivariate adjustment for body mass index and other coronary risk factors only modestly attenuated the RRs. Compared with nondiabetic persons, the multivariate RRs of fatal CHD across categories of diabetes duration (< or =5, 6-10, 11-15, 16 25, >25 years) were 2.75, 3.63, 5.51, 6.38, and 11.9 (P< .001 for trend), respectively. The combination of prior CHD and a long duration of clinical diabetes (ie, >15 years) was associated with a 30-fold (95% CI, 20.7-43.5) increased risk of fatal CHD. CONCLUSIONS: Our data indicate that among women, history of diabetes is associated with dramatically increased risks of death from all causes and fatal CHD. The combination of diabetes and prior CHD identifies particularly high-risk women. PMID- 11485505 TI - Depression and risk of heart failure among older persons with isolated systolic hypertension. AB - BACKGROUND: Investigators have shown that depression is associated with an increased risk of coronary heart disease in general and myocardial infarction in particular. However, it is unknown whether depression, independent of its association with myocardial infarction, is a risk factor for heart failure. METHODS: This study examined whether depression was a predictor of incident heart failure among 4538 persons aged 60 years and older with isolated systolic hypertension who were enrolled in the Systolic Hypertension in the Elderly Program (SHEP). Depression was defined as a score of 16 or more at baseline on the Center for Epidemiological Studies Depression Scale (CES-D). The relationship between depression and heart failure was assessed using Cox proportional hazards regression. RESULTS: The average follow-up was 4.5 years. Heart failure developed in 138 (3.2%) of 4317 nondepressed persons and in 18 (8.1%) of 221 depressed persons. After controlling for age; sex; race; history of myocardial infarction, diabetes, or angina; blood pressure; cholesterol levels; electrocardiographic abnormalities; smoking; disability; and SHEP treatment group, depressed persons had more than a 2-fold higher risk of developing heart failure compared with nondepressed persons (hazard ratio, 2.59; 95% confidence interval, 1.57-4.27; P<.001). After additional adjustment for the occurrence of myocardial infarction during follow-up, depressed persons remained at elevated risk of heart failure (hazard ratio, 2.82; 95% confidence interval, 1.71-4.67; P<.001). CONCLUSIONS: Depression is independently associated with a substantial increase in the risk of heart failure among older persons with isolated systolic hypertension. This association does not appear to be mediated by myocardial infarction. PMID- 11485506 TI - A randomized controlled clinical trial of the serotonin type 3 receptor antagonist alosetron in women with diarrhea-predominant irritable bowel syndrome. AB - BACKGROUND: Irritable bowel syndrome (IBS) is a common gastrointestinal disorder seen in primary care practice. The symptoms of IBS, including abdominal pain, discomfort, and abnormal bowel function, may be modulated by activity of the serotonin type 3 receptor (5-HT(3)). The efficacy and tolerability of the 5-HT(3) receptor antagonist alosetron hydrochloride in nonconstipated female patients with IBS were evaluated in a double-blind, randomized, placebo-controlled trial. METHODS: Patients received either 1 mg of alosetron hydrochloride (n = 309) or placebo (n = 317) twice daily for 12 weeks, followed by a 4-week posttreatment period. Adequate relief of IBS pain and discomfort was the primary end point. Secondary end points included improvements in urgency, stool frequency, stool consistency, incomplete evacuation, and bloating. RESULTS: Seventy-one percent of patients were classified as having diarrhea-predominant IBS. Forty-three percent of alosetron-treated patients with diarrhea-predominant IBS reported adequate relief for all 3 months compared with 26% of placebo-treated patients (P<.001; percentage point difference = 17; 95% confidence interval, 8.0-25.4). Improvement with alosetron compared with placebo was observed by the end of the fourth week of treatment and persisted throughout the remainder of treatment. Alosetron significantly decreased urgency and stool frequency and caused firmer stools within 1 week of starting treatment. Effects were sustained throughout treatment and symptoms returned following treatment cessation. No significant improvement in the percentage of days with sense of incomplete evacuation or bloating was observed compared with placebo during the first month of treatment. Constipation was the most commonly reported adverse event. CONCLUSION: Alosetron hydrochloride, 1 mg twice daily for 12 weeks, is effective in relieving pain and some bowel-related symptoms in diarrhea-predominant female patients with IBS. PMID- 11485507 TI - Pneumococcal peritonitis in adult patients: report of 64 cases with special reference to emergence of antibiotic resistance. AB - BACKGROUND: Few data are available regarding pneumococcal peritonitis. We studied the clinical characteristics of intra-abdominal infections caused by Streptococcus pneumoniae and its prognosis in relation to antibiotic resistance. METHODS: We reviewed all cases of culture-proved pneumococcal peritonitis. Patients with liver cirrhosis and primary pneumococcal peritonitis were compared with patients with Escherichia coli peritonitis. RESULTS: Between January 1, 1979, and December 31, 1998, we identified 45 cases of primary pneumococcal peritonitis in patients with cirrhosis and 19 cases of secondary (or tertiary) pneumococcal peritonitis. Patients with cirrhosis and primary pneumococcal peritonitis vs those with primary E coli peritonitis had more frequent community acquired infection, 73% vs 47%; pneumonia, 36% vs 2%; and bacteremia, 76% vs 33%; and higher attributable mortality (early mortality), 27% vs 9% (P<.05 for all). Secondary (or tertiary) pneumococcal peritonitis was associated with upper or lower gastrointestinal tract diseases; in most cases, the infection appeared after surgery. A hematogenous spread of S pneumoniae from a respiratory tract infection might be the most important origin of peritonitis; also, S pneumoniae might directly reach the gastrointestinal tract favored by endoscopic procedures or hypochlorhydria. There was an increased prevalence of penicillin and cephalosporin resistance up to 30.7% and 17.0%, respectively, although it was not associated with increased mortality rates. CONCLUSIONS: Primary pneumococcal peritonitis in patients with cirrhosis more often spread hematogenously from the respiratory tract and was associated with early mortality. In secondary (and tertiary) pneumococcal peritonitis, a transient gastrointestinal tract colonization and inoculation during surgery might be the most important mechanisms. Current levels of resistance were not associated with increased mortality rates. PMID- 11485508 TI - Outcome of adult cardiopulmonary resuscitations at a tertiary referral center including results of "limited" resuscitations. AB - BACKGROUND: The results of in-hospital resuscitations may depend on a variety of factors related to the patient, the environment, and the extent of resuscitation efforts. We studied these factors in a large tertiary referral hospital with a dedicated certified resuscitation team responding to all cardiac arrests. METHODS: Statistical analysis of 445 prospectively recorded resuscitation records of patients who experienced cardiac arrest and received advanced cardiac life support resuscitation. We also report the outcomes of an additional 37 patients who received limited resuscitation efforts because of advance directives prohibiting tracheal intubation, chest compressions, or both. MAIN OUTCOME MEASURES: Survival immediately after resuscitation, at 24 hours, at 48 hours, and until hospital discharge. RESULTS: Overall, 104 (23%) of 445 patients who received full advanced cardiac life support survived to hospital discharge. Survival was highest for patients with primary cardiac disease (30%), followed by those with infectious diseases (15%), with only 8% of patients with end-stage diseases surviving to hospital discharge. Neither sex nor age affected survival. Longer resuscitations, increased epinephrine and atropine administration, multiple defibrillations, and multiple arrhythmias were all associated with poor survival. Patients who experienced arrests on a nursing unit or intensive care unit had better survival rates than those in other hospital locations. Survival for witnessed arrests (25%) was significantly better than for nonwitnessed arrests (7%) (P =.005). There was a disproportionately high incidence of nonwitnessed arrests during the night (12 AM to 6 AM) in unmonitored beds, resulting in uniformly poor survival to hospital discharge (0%). None of the patients whose advance directives limited resuscitation survived. CONCLUSIONS: Very ill patients in unmonitored beds are at increased risk for a nonwitnessed cardiac arrest and poor resuscitation outcome during the night. Closer vigilance of these patients at night is warranted. The outcome of limited resuscitation efforts is very poor. PMID- 11485509 TI - The effect of BCG vaccination on tuberculin reactivity and the booster effect among hospital employees. AB - BACKGROUND: We estimated the effect of remote BCG vaccination on tuberculin reactivity and the booster effect among hospital employees. METHODS: Cross sectional survey at a university hospital. All personnel employed during a 24 month period were included in the study. Employees were administered 2-step tuberculin testing, and BCG vaccination scars were verified. RESULTS: Of 665 hospital employees studied, 239 (36%) had been vaccinated with BCG in childhood. Significant tuberculin reactions (> or =5 mm) were more frequent among BCG vaccinated (60%) than among nonvaccinated (29%) employees (odds ratio [OR], 3.6; 95% confidence interval [CI], 2.6-5.2). The predictive value of tuberculosis infection increased with increasing reaction size and greater age (from 37% in subjects 30 years or younger with indurations > or =5 mm to 100% in subjects 50 years or older with indurations > or =15 mm). Among 374 employees with a negative tuberculin test reaction who underwent a second test, 39 (43%) of 91 vaccinated subjects had a positive booster reaction in contrast to 51 (22%) of 232 nonvaccinated subjects (OR, 3.4; 95% CI, 2-5.7). Neither different size criteria nor different definitions of the booster effect had an impact on the predictive value of tuberculosis infection. CONCLUSIONS: Remote BCG vaccination largely influences the tuberculin reaction and the boosting phenomenon among hospital employees. The interpretation of the results of 2-step tuberculin testing in a BCG-vaccinated subject must take into account age, size of the reaction, and local prevalence of tuberculosis infection. No single criterion, however, can accurately separate reactions caused by true infection from those caused by BCG vaccination. PMID- 11485510 TI - Diabetes and depot medroxyprogesterone contraception in Navajo women. AB - BACKGROUND: Depot medroxyprogesterone acetate contraception is widely used in Navajo women, a high-risk population for diabetes mellitus. However, depot medroxyprogesterone may lead to weight gain and independently decrease insulin sensitivity. We studied the association between depot medroxyprogesterone and development of diabetes in Navajo women. METHODS: We studied Navajo women aged 18 to 50 years who had seen a health care provider at a Navajo Area Indian Health Service clinic at least once in 1998. Diabetic cases (n = 284) and nondiabetic controls (n = 570) were matched by age. Medical records were reviewed to determine contraception use before the diagnosis date of diabetes. RESULTS: Users of depot medroxyprogesterone were more likely to develop diabetes than patients who had used combination estrogen-progestin oral contraception only (odds ratio [OR], 3.8; 95% confidence interval [CI], 1.8-7.9). The excess risk persisted after adjustment for body mass index (OR, 3.6; 95% CI, 1.6-7.9). Longer use was associated with greater risk of diabetes. Users of depot medroxyprogesterone were also more likely to develop diabetes than patients who had never used hormonal contraception, although excess risk was smaller (OR, 2.4; 95% CI, 1.4-3.6). CONCLUSIONS: Depot medroxyprogesterone contraception was associated with a greater risk of diabetes compared with combination oral contraceptive use only. Risk was associated with length of use and persisted after adjustment for body mass index. Additional research is needed for confirmation, but this risk should be considered in contraceptive choice for women at high risk for diabetes. PMID- 11485512 TI - What is proper treatment for Wegener granulomatosis? PMID- 11485511 TI - The effect of hormone replacement therapy on cardiovascular risk factors in type 2 diabetes: a randomized controlled trial. AB - BACKGROUND: Postmenopausal women with diabetes are at high risk for cardiovascular disease, compared with their nondiabetic counterparts. Combined continuous hormone replacement therapy (HRT) is associated with improvements in serum lipoprotein levels in nondiabetic women; however, the effect in women with diabetes has not been determined. We evaluated the effect of combined continuous HRT on lipoprotein and coagulation factor concentrations and glycemic control in postmenopausal women with type 2 diabetes mellitus. METHODS: In a randomized controlled crossover study, 61 subjects received combined continuous HRT or placebo. Each treatment phase was of 6 months' duration, with an 8-week washout phase between treatment phases. RESULTS: Total cholesterol concentration decreased by 7% (95% confidence interval [CI], 4%-11%) during HRT. For low density lipoprotein concentration, the mean decrease with HRT was 12% (95% CI, 6% 17%). Apolipoprotein B levels decreased in keeping with the reduction in low density lipoprotein cholesterol concentrations. There were no significant changes in concentrations of high-density lipoprotein, its subfractions, or triglycerides. Lipoprotein(a) and fibrinogen concentrations were reduced by 21% (95% CI, 10%-31%) and 8% (95% CI, 2%-13%), respectively, with HRT. Fructosamine concentrations declined by 5% (95% CI, 2%-9%) during HRT. CONCLUSIONS: In postmenopausal women with type 2 diabetes mellitus, combined continuous HRT has beneficial effects on lipoprotein concentrations and improves some markers of coagulation and glycemic control. PMID- 11485513 TI - Acute thrombocytopenic purpura in a patient treated with chlordiazepoxide and clidinium. PMID- 11485514 TI - Severe bradycardia after a methylprednisolone "minipulse" treatment. PMID- 11485515 TI - Considerations in diagnosis of Cushing syndrome. PMID- 11485517 TI - Osteoporosis and fracture risk prevention in long-term glucocorticoid therapy. PMID- 11485519 TI - The surgeon as professional: a challenge to our educators. PMID- 11485520 TI - Safety and efficacy of radiofrequency thermal ablation in advanced liver tumors. AB - HYPOTHESIS: Radiofrequency thermal ablation (RFA) can be performed safely and effectively to control local disease in patients with advanced, unresectable liver tumors. DESIGN, SETTING, AND PATIENTS: Prospective study of 76 patients with unresectable liver tumors who underwent RFA at a private tertiary referral hospital. INTERVENTIONS: Ninety-nine RFA operations were performed to ablate 328 tumors. MAIN OUTCOME MEASURES: Complications and local recurrence. RESULTS: There was 1 death (1%), major complications occurred in 7 operations (7%), and minor complications occurred in 10 operations (10%). Local recurrence was identified in 30 tumors (9%) at a mean follow-up of 15 months. Size (P<.001), vascular invasion (P<.001), and total volume ablated (P<.001) were associated with recurrence but the number of tumors was not (P =.39). CONCLUSION: Radiofrequency thermal ablation provides local control of advanced liver tumors with low recurrence and acceptable morbidity. PMID- 11485521 TI - Laparoscopic Heller myotomy and Dor fundoplication for achalasia: analysis of successes and failures. AB - BACKGROUND: In the treatment of achalasia, surgery has been traditionally reserved for patients with residual dysphagia after pneumatic dilatation. The results of laparoscopic Heller myotomy have proven to be so good, however, that most experts now consider surgery the primary treatment. HYPOTHESIS: The outcome of laparoscopic myotomy and fundoplication for achalasia is dictated by technical factors. SETTING: University hospital tertiary care center. DESIGN: Retrospective study. PATIENTS AND METHODS: One hundred two patients with esophageal achalasia underwent laparoscopic Heller myotomy and Dor fundoplication. Fifty-seven patients had been previously treated by pneumatic dilatation or botulinum toxin. The design of the operation involved a 7-cm myotomy, which extended 1.5 cm onto the gastric wall, and a Dor fundoplication. Esophagrams, esophageal manometric findings, and video records of the procedure were analyzed to determine the technical factors that contributed to the clinical success or failure of the operation. MAIN OUTCOME MEASURE: Swallowing status. RESULTS: In 91 (89%) of the 102 patients, good or excellent results were obtained after the first operation. A second operation was performed in 5 patients to either lengthen the myotomy (3 patients) or take down the fundoplication (2 patients). Dysphagia resolved in 4 of these patients. The remaining 6 patients were treated by pneumatic dilatation, but dysphagia improved in only 1. At the conclusion of treatment, excellent or good results had been obtained in 96 (94%) of the 102 patients. CONCLUSIONS: These data show that a Heller myotomy was unsuccessful in patients with an esophageal stricture; a short myotomy and a constricting Dor fundoplication were the avoidable causes of residual dysphagia; a second operation, but not pneumatic dilatation, was able to correct most failures; and that the identified technical flaws were eliminated from the last half of the patients in the series. PMID- 11485522 TI - Localization and reoperation results for persistent and recurrent parathyroid carcinoma. AB - HYPOTHESIS: Reoperation is safe and benefits patients with persistent and recurrent parathyroid carcinoma. DESIGN: Retrospective study. The mean follow-up time was 8.1 years (median, 7 years; range, 1-23 years). SETTING: A university tertiary referral center. PATIENTS: Eighteen patients treated for parathyroid carcinoma from 1966 to 1999. RESULTS: The mean serum calcium level was 13.7 mg/dL (3.43 mmol/L), and the parathyroid hormone (PTH) level was 1.6 to 20 times the upper limit of normal. Fourteen of 18 patients had persistent or recurrent parathyroid carcinoma and underwent 54 reoperations (28 at our institution). Mean time to recurrence was 4.8 years (range, 1-20 years). Symptoms of hyperparathyroidism were relieved in 86% of patients who had reoperation (P<.05). Reoperation for parathyroid carcinoma (25 locoregional and 3 distant) significantly reduced and normalized the serum calcium and PTH levels in 75% and 62% of the cases, respectively (P<.001). The preoperative serum calcium level was a significant predictor of postreoperative normalization of the serum calcium level but not extent of initial resection, PTH level, time to recurrence, concordance of localization studies, or patient age and sex (P<.01). Surgical complications consisted of 5 unilateral and 1 bilateral permanent recurrent laryngeal nerve palsies (2 intentionally resected en bloc), 1 transient hypoparathyroidism, 1 wound seroma, and 1 tracheoesophageal fistula. The sensitivity rates of sestamibi scan (n = 14), magnetic resonance imaging (n = 15), computed tomographic scan (n = 6), ultrasound (n = 13), and selective venous catheterization with PTH measurement (n = 6) were 79%, 93%, 67%, 69%, and 83%, respectively. CONCLUSIONS: Recurrence is common in patients with parathyroid carcinoma. Patients with this disease should have frequent, lifelong follow-up to ensure early detection of recurrence. Although reoperation for persistent or recurrent parathyroid carcinoma provides significant symptomatic relief and normalizes serum calcium and PTH levels in most patients, it is associated with some morbidity. Localizing studies of parathyroid carcinoma are helpful but do not detect all tumor foci. PMID- 11485523 TI - The evolving treatment of anal cancer: How are we doing? AB - HYPOTHESIS: The adaptation of new techniques in treatment of epidermoid carcinoma of the anal canal during the past 3 decades has improved clinical outcomes. DESIGN: Retrospective consecutive case review. SETTING: A university hospital and Veterans Affairs medical center. PATIENTS: Medical records of 76 consecutive patients treated for invasive epidermoid cancer of the anal canal between 1970 and 1999 were reviewed. Twenty-one patients were excluded because of inadequate staging information and/or follow-up of less than 12 months. MAIN OUTCOME MEASURES: Locoregional recurrence, survival, colostomy-free survival, and morbidity. RESULTS: Fifty-five patients composed the study population. Ten were treated during decade 1 (1970-1979), 16 in decade 2 (1980-1989), and 29 in decade 3 (1990-1999). Mean age and sex distributions were similar. The prevailing primary treatment modality changed during the course of the study from sequential treatment (chemotherapy then radiation therapy then radical surgery) to concurrent chemoradiation (70% and 0% of cases, respectively, in decade 1 to 7% and 76% of cases, respectively, in decade 3). Locoregional control (50%, 81%, and 93%; P =.01), crude survival (median, 28, 30, and 76 months), and colostomy-free survival (mean, 13, 90, and 80 months) improved during the 3 decades. There were no differences in major complications during the 3 decades (40%, 56%, and 41%). CONCLUSION: Primary treatment with concurrent chemoradiation has improved the local recurrence, survival, and colostomy-free survival rates in patients with invasive epidermoid carcinoma of the anal canal without increasing major morbidity. PMID- 11485524 TI - Laparoscopic Heller myotomy improves esophageal emptying and the symptoms of achalasia. AB - HYPOTHESIS: Laparoscopic Heller esophageal myotomy improves esophageal clearance and symptoms of achalasia in the early and late postoperative periods. DESIGN: We followed up 98 consecutive patients attending a referral center between February 1, 1994, and July 1, 2000, who underwent laparoscopic myotomy. Operative time, complications, and length of stay were recorded. Postoperative outcomes were assessed using Van Trappen symptom scores (1 indicates no symptoms; 2, symptoms occurring less than once a week; 3, symptoms occurring more than once weekly; and 4, persistent symptoms) and scintigraphic esophageal transit studies. RESULTS: Of 98 patients, 91 underwent anterior fundoplication. There were no open conversions and 1 mucosal perforation, which was closed laparoscopically without complications. Mean operative times and postoperative days were 3.2 hours and 4.3 days, respectively, in the first 32 patients and 1.7 hours and 2.3 days, respectively, in the last 32 patients (P<.001). Postoperative complications included pneumothorax (4% of patients), atelectasis (5%), and delayed gastric emptying (1%). Seventy-five percent of patients gained weight after surgery. At longest follow-up, 91% of patients were satisfied with the outcome of the procedure. Mean Van Trappen scores for dysphagia improved from 4.0 in the preoperative period to 1.2 at early and late follow-up (P<.001). Fluid retention at 10 minutes in the upright position was 47% in the preoperative period and improved at early and late follow-up to 21% and 20%, respectively (P<.001). CONCLUSIONS: Laparoscopic Heller myotomy can safely reverse the symptoms of achalasia and improve esophageal transit. These benefits, realized during the early postoperative period, were maintained at longest follow-up. PMID- 11485525 TI - Increased transplantation of kidneys with multiple renal arteries in the laparoscopic live donor nephrectomy era: surgical technique and surgical and nonsurgical donor and recipient outcomes. AB - BACKGROUND: For anatomical and technical reasons, many transplant centers restrict laparoscopic live donor nephrectomy (in contrast with open live donor nephrectomy) to left kidneys. HYPOTHESIS: This change in surgical practice increases procurement and transplantation rates of live donor kidneys with multiple renal arteries (RAs), without affecting donor and recipient outcomes. DESIGN AND SETTING: Retrospective review at an academic tertiary care referral center comparing laparoscopically procured single vs multiple-RA kidney grafts (April 1997 to October 2000). PATIENTS: Seventy-nine consecutive left laparoscopic live kidney donors and 78 transplant recipients. MAIN OUTCOME MEASURES: Donor and recipient complications and postoperative length of stay; cold and warm ischemia time; operating time; short-term and long-term graft function; and survival. RESULTS: We noted multiple RAs in 21 (27%) of all kidneys. The proportion of donors with 1 or more perioperative complications was 19% in the single-RA group vs 10% in the multiple-RA group (P was not significant). For the recipients, we noted no significant differences between groups with respect to surgical complications, quality of early and late graft function, rejection rates, graft losses (all immunologic), and graft survival. Cold and warm ischemia time and length of stay were similar for donors and recipients in both groups. Median operating times were significantly longer for the multiple-RA vs single-RA group (difference, 41 minutes for donors and 45 minutes for recipients; P<.02). CONCLUSIONS: While the introduction of laparoscopic live donor nephrectomy has significantly increased the number of grafts with multiple RAs (compared with historical open controls), this change in practice is safe for both donors and recipients from a patient outcome-based perspective. However, from an economic perspective, the longer operating time associated with multiple-RA grafts provides strong added rationale for optimization of surgical instruments and techniques to make right-sided laparoscopic nephrectomy a routine intervention. PMID- 11485526 TI - Systemic coagulation and fibrinolysis after laparoscopic and open gastric bypass. AB - HYPOTHESIS: Laparoscopic gastric bypass (GBP) induces a postoperative hypercoagulable state that is similar or reduced compared with open GBP. SETTING: University hospital. PATIENTS: Between May 1999 and June 2000, 70 patients were randomly assigned to laparoscopic (n = 36) or open (n = 34) GBP. Deep venous thrombosis (DVT) prophylaxis consisted of antiembolism stockings and sequential pneumatic compression devices. MAIN OUTCOME MEASURES: Plasminogen, thrombin antithrombin complex (TAT), prothrombin fragment 1.2 (F1.2), fibrinogen, D-dimer, antithrombin III (AT), and protein C levels were measured at baseline and at 1, 24, 48, and 72 hours postoperatively. A venous duplex examination of both lower extremities was performed preoperatively and between the third and fifth day postoperatively. RESULTS: The 2 groups were similar in age, weight, and body mass index. Plasminogen levels decreased, and TAT, F1.2, and fibrinogen levels increased after laparoscopic and open GBP. There was no significant difference in these levels between groups. D-dimer levels increased in both groups, but the levels were significantly higher after open GBP than after laparoscopic GBP (P<.01). Antithrombin III and protein C levels decreased in both groups. The reduction of AT (at 1 hour) and protein C (at 72 hours) was significantly less after laparoscopic GBP than after open GBP (P<.05). Postoperative venous duplex examination revealed DVT in 1 (2.9%) of 34 patients after open GBP but in none of 36 patients after laparoscopic GBP. One patient developed pulmonary embolism after open GBP. CONCLUSIONS: Laparoscopic GBP induces a hypercoagulable state similar to that of open GBP. Our findings suggest that DVT prophylaxis should be used during laparoscopic GBP as in open GBP. PMID- 11485527 TI - Factors determining convalescence after uncomplicated laparoscopic cholecystectomy. AB - HYPOTHESIS: Detailed information on duration and limiting factors for convalescence after uncomplicated laparoscopic cholecystectomy is lacking. Duration of convalescence may be associated with patients' expectations, given recommendations, and postoperative complaints such as pain and fatigue. DESIGN: Prospective, descriptive study. SETTING: A university hospital. PATIENTS: Two hundred consecutive patients who underwent uncomplicated elective laparoscopic cholecystectomy. INTERVENTION: For sedentary, light, or moderate workload or main recreational activity, we recommended 2 days of postoperative convalescence; for strenuous workload or recreational activity, we recommended 1 week. MAIN OUTCOME MEASURES: Duration and reasons for absence from work. RESULTS: Convalescence from work (n = 85) and recreational activity (n = 198) was 6 days (range, 0-28 days) and 2 days (range, 0-24 days), respectively, in patients recommended for 2 days' convalescence. In patients recommended for 1 week of convalescence, convalescence from work (n = 25) was 10 days (range, 0-52 days), and convalescence from main recreational activity (n = 2), 8 days (range, 5-11 days). Among 87 patients who resumed work or activity later than recommended, pain was a contributory cause in 41 patients, fatigue in 35 patients, and convalescent period falling on a weekend in 26 patients, while 29 patients had arranged vacation or sick leave preoperatively. Preoperative expectation of convalescence and pain were independent contributory factors (P<.01) for convalescence from work for longer than 2 days in patients recommended for 2 days' convalescence. CONCLUSIONS: The period of convalescence after uncomplicated laparoscopic cholecystectomy is about 1 week from work and 2 days from recreational activity when 2 days of convalescence is recommended. Improved pain relief and patient information may further reduce convalescence. PMID- 11485528 TI - Randomized comparison of ultrasonic vs clamp transection of the liver. AB - HYPOTHESIS: Hepatic parenchymal transection is a technical priority in liver surgery. The use of an ultrasonic dissector for hepatectomy may result in less blood loss than conventional clamp crushing. DESIGN: Randomized controlled trial. SETTING: University teaching hospital. PATIENTS: The 132 patients scheduled to undergo partial hepatectomies were randomly assigned to receive hepatic transection by ultrasonic dissector or by clamp crushing (66 patients by each method). INTERVENTIONS: All resections were performed with inflow occlusion and were guided ultrasonographically. Hepatectomies were graded according to a predefined system based on 6 criteria (blood loss, transection time, technical error, surgical margin, landmark appearance, and postoperative morbidity), each with 3 scores (lower scores indicating higher quality). MAIN OUTCOME MEASURES: Blood loss and hepatectomy grade. RESULTS: No difference was found between the ultrasonic and clamp groups in median blood loss (515 mL [range, 15-2527 mL] vs 452 mL [range, 17-1912 mL]; P =.63), transection time (61 minutes [range, 16-177 minutes] vs 54 minutes [range, 7-205 minutes]; P =.58), or transection speed (1.1 cm(2)/min [range, 0.4-4.0 cm(2)/min] vs 1.0 cm(2)/min [range, 0.4-3.0 cm(2)/min]; P =.90). Ultrasonic dissection caused more frequent histologically proven tumor exposure at the surgical margin (9 vs 3 patients; P =.09), incomplete appearance of landmark hepatic veins on the cut surface after anatomical resection (12 vs 4 patients; P =.03), and postoperative morbidity (20 vs 14 patients; P =.32) than did clamp crushing. The hepatectomies with clamp crushing had significantly higher grades than those with ultrasonic dissection (P =.05), as indicated by the lower median sum score (4.0 [range, 0-12] vs 5.0 [range, 0-19]; 95% confidence interval for difference, -2.0 to 0; P =.03). The transection method independently influenced hepatectomy grade (adjusted odds ratio = 3.06; 95% confidence interval, 1.35-6.92; P =.01). CONCLUSIONS: Ultrasonic dissection offers no reduction in blood loss compared with clamp crushing for transection of the liver. Clamp crushing results in a higher quality of hepatectomy and is therefore the option of choice. PMID- 11485530 TI - Delayed superficialization of brachiobasilic fistula: technique and initial experience. AB - HYPOTHESIS: Angioaccess procedures for dialysis have varied patency rates with frequent need for intervention. A superficialized arteriovenous brachiobasilic fistula created as a 2-step procedure will have good long-term patency with minimal complication. DESIGN: Retrospective medical record review and patient interview. SETTING: Tertiary referral university hospital. PATIENTS: Twelve patients who underwent delayed superficialization of brachiobasilic fistula from September 1994 to April 2000. MAIN OUTCOME MEASURES: Patency of fistula for dialysis, and major and minor complications, including revisions. RESULTS: Delayed superficialization of brachiobasilic fistula was performed in 12 patients. Fistulas have been used for a mean duration of 22.4 months (range, 10 59 months). Two patients required alternate access owing to thrombosis of brachiobasilic fistula. CONCLUSIONS: The delayed superficialized brachiobasilic arteriovenous fistula has a good initial patency rate with minimal complications. It should be considered early in patients if radiocephalic fistula is unavailable. PMID- 11485531 TI - Enteral vs parenteral nutrition after major abdominal surgery: an even match. AB - HYPOTHESIS: Immediate enteral feeding following major abdominal surgery reduces postoperative complications and mortality when compared with parenteral nutrition. DESIGN: A prospective multicenter randomized trial. SETTING: A university hospital department of digestive surgery. PATIENTS AND INTERVENTIONS: Two hundred forty-one malnourished patients undergoing major elective abdominal surgery were randomly assigned to receive, after surgery, either enteral (enteral nutrition group: 119 patients) or parenteral nutrition (total parenteral nutrition group: 122 patients). The patients were monitored for postoperative complications and mortality. RESULTS: The rate of major postoperative complications was similar in the enteral and parenteral groups (enteral nutrition group: 37.8%; total parenteral nutrition group: 39.3%; P was not significant), as were the overall postoperative mortality rates (5.9% and 2.5%, respectively; P was not significant). CONCLUSION: The present study failed to demonstrate that enteral feeding following major abdominal surgery reduces postoperative complications and mortality when compared with parenteral nutrition. PMID- 11485532 TI - Serum levels of transforming growth factor beta1 in patients with breast cancer. AB - HYPOTHESIS: Transforming growth factor beta1 (TGF-beta1) may be related to breast cancer progression. DESIGN: Prospective study. SETTING: University hospital. PATIENTS: Sixty consecutive patients with invasive breast cancer undergoing surgery were prospectively included and evaluated. The control group consisted of 14 patients with benign breast tumors (7 with fibrocystic disease and 7 with fibroadenoma). INTERVENTION: Venous blood samples were collected before the surgery. Sera were obtained by centrifugation and stored at -70 degrees C until assayed. Serum concentrations of TGF-beta1 were measured by quantitative sandwich enzyme immunoassay. Data on primary tumor stage, age, estrogen receptor status, lymph node status, distant metastases, and TNM staging (according to the Union Internationale Contre le Cancer) were reviewed and recorded. MAIN OUTCOME MEASURES: Measurements of preoperative serum TGF-beta1 levels in patients with breast cancer. RESULTS: The mean +/- SD value of serum TGF-beta1 in patients with invasive breast cancer was 498.7 +/- 249.7 pg/mL and in the control group was 495.2 +/- 225.5 pg/mL (P =.96). However, there were significantly higher serum levels of TGF-beta1 in patients with more advanced lymph node status (P =.04), more advanced TNM stage (P =.005), and poorer histological grade (P =.02). In multivariate analysis, TNM staging (P =.02) was demonstrated to be the independent factor related to significantly higher serum levels of TGF-beta1. CONCLUSIONS: Patients with more advanced TNM stages were shown to have higher serum TGF-beta1 levels. Thus, serum TGF-beta1 levels may reflect the severity of invasive breast cancer. PMID- 11485534 TI - Multivariate analysis of clinicopathologic parameters for the insular subtype of differentiated thyroid carcinoma. AB - HYPOTHESIS: Insular carcinoma represents a more aggressive subtype of differentiated thyroid cancer on multivariate analysis after controlling for various clinicopathologic parameters. DESIGN: Retrospective analysis. SETTING: Tertiary referral center at a university hospital. PATIENTS: One hundred twenty seven consecutive patients having a histological diagnosis of the follicular variant of papillary thyroid carcinoma or follicular thyroid carcinoma. MAIN OUTCOME MEASURE: A logistic regression model was used to examine the relationship between various clinicopathologic parameters and the insular subtype. RESULTS: The insular subtype involved 14 of 127 tumors. Unlike extrathyroidal extension and nodal metastasis, primary tumor diameter (> 40 mm vs < or = 40 mm; P = .008) and distant metastasis (P = .003) correlated with the insular subtype. Both parameters were interrelated since tumors greater than 40 mm displayed distant metastasis more often (30% vs 8%; P = .008) than tumors measuring 40 mm or less. CONCLUSIONS: These findings suggest that an unidentified somatic event may induce an accelerated proliferation of the transformed thyrocytes, which may ultimately result in enhanced rates of distant metastasis with increasing tumor volume. PMID- 11485536 TI - A spike in parathyroid hormone during neck exploration may cause a false-negative intraoperative assay result. AB - HYPOTHESIS: We hypothesize that false-negative results using the rapid intraoperative parathyroid hormone (IOPTH) assay can be caused by spikes in the level of parathyroid hormone that occur during mobilization of the adenoma. DESIGN: Retrospective analysis of a case series. SETTING: University tertiary care center. PATIENTS: Ten consecutive patients with primary hyperparathyroidism. INTERVENTIONS: All patients underwent neck exploration with IOPTH monitoring. Using a sampling protocol described in the literature, IOPTH values were checked at the time of incision, during mobilization of the adenoma, and 10 minutes after resection of the adenoma. MAIN OUTCOME MEASURES: Patients were evaluated for adequate parathyroid tissue excision as determined by IOPTH levels and examination of ipsilateral glands. All patients had normal serum calcium values documented postoperatively. Parathyroid hormone half-life was calculated assuming first-order kinetic decay. RESULTS: Nine patients had an appropriate decline in IOPTH with a mean +/- SD parathyroid hormone half-life of 3.9 +/- 1.08 minutes. Mobilization of the adenoma resulted in a spike in the IOPTH value, with 1 patient's value increasing from a baseline of 95.5 pg/mL (10.1 pmol/L) to 751 pg/mL (79.1 pmol/L). Another patient who was confirmed to have a solitary adenoma had a false-negative postexcision value. A spike in IOPTH that occurred during neck dissection was not detected by the sampling protocol and explains the false negative value. A literature review revealed that most protocols check baseline values early in the operation and are at risk for false-negative results due to a spike from mobilization of the adenoma. CONCLUSIONS: These data demonstrate that false-negative IOPTH assay findings can result from a spike in parathyroid hormone level during exploration, which may go unrecognized if baseline values are measured during the early stages of mobilization of the adenoma. We have altered our assay protocol and have begun measuring IOPTH at the time of neck incision, at the time the adenoma is completely removed (time zero [t(0)]), and 10 minutes after excision. PMID- 11485537 TI - Surgical resection for metastatic melanoma to the liver: the John Wayne Cancer Institute and Sydney Melanoma Unit experience. AB - HYPOTHESIS: Metastatic melanoma to the liver is not incurable; complete surgical resection can achieve long-term survival in selected patients. BACKGROUND: Metastases to the liver are diagnosed in 10% to 20% of patients with American Joint Committee on Cancer stage IV melanoma. Surgical resection has not been generally accepted as a therapeutic option, as most patients will have other sites of disease that limit their survival to a median of only 4 to 6 months. However, there is little information on outcomes following resection in those patients with disease limited to the liver. PATIENTS AND METHODS: Review of the prospective melanoma databases at the John Wayne Cancer Institute, Santa Monica, Calif, and the Sydney Melanoma Unit, Sydney, Australia, identified 1750 patients with hepatic metastases, of whom 34 (2%) underwent exploration with intent to resect the metastases. Prognostic factors within the group of patients who underwent resection were examined by univariate and multivariate analysis, and median disease-free survival (DFS) and overall survival (OS) were calculated. RESULTS: Of 34 patients undergoing exploratory celiotomy, 24 (71%) underwent hepatic resection and 10 (29%) underwent exploration but not resection. Eighteen patients (75%) underwent complete surgical resection, while the remaining 6 underwent palliative or debulking procedures with incomplete resection. The operative resections included lobectomy (n=14), segmentectomy (4), nonanatomic resection (5), and extended lobectomy (1). The median number of resected lesions was 1, and median lesion size was 5 cm (range, 0.7-22 cm). The median disease free interval between initial diagnosis of melanoma and development of hepatic metastases was 58 months (range, 0-264 months). Median DFS and OS estimates in the 24 patients who underwent surgical resection were 12 months (range, 0-147 months) and 28 months (range, 2-147 months), respectively. Five-year DFS and OS in this group were 12% and 29%. Macroscopically, complete resection of disease (P =.001) and histologically negative resection margins (P =.03) significantly improved DFS by univariate analysis. Patients rendered surgically free of disease also tended to have improved OS (P =.06). Median OS was 28 months for patients who underwent surgical resection compared with 4 months for patients who underwent exploration only (P<.001). CONCLUSIONS: Resection of metastatic melanoma to the liver may improve DFS and OS in selected patients, similar to resection of other metastatic sites. Therefore, patients with limited metastatic sites, including the liver, who can be rendered free of disease should be considered for complete surgical resection, as their prognosis is otherwise dismal. PMID- 11485539 TI - Ex situ splitting of the liver: the versatile Paul Brousse technique. AB - Split liver transplantation was introduced to alleviate the organ shortage. Comparable morbidity, graft, and patient survival rates have been reported with the use of either split or whole liver for transplantation. This article gives a detailed illustrated report of our splitting technique. Alternative techniques are also discussed. Both techniques of in situ vs ex situ splitting should be developed according to the logistical possibilities. Depending on the desired volume of the left graft, the cutting line may be shifted, particularly to the right of the middle hepatic vein, the whole of segment 4 being then included in the left graft (when the recipients are 2 adults). Split liver transplantation increases graft availability by 15% to 28%. At present, the goal of split liver transplantation clearly has to be its clinical application in 2 adults to tackle the problem of organ shortage in this population, which accounts for more than 90% of deaths in those who are on the waiting list. PMID- 11485540 TI - Image of the month: Achalasia. PMID- 11485541 TI - Military damage control. PMID- 11485543 TI - Managing hepatic bleeding with autologous plasma/collagen-based fibrin sealant. PMID- 11485544 TI - Moments in surgical history: Philip Syng Physick (1768-1837). PMID- 11485545 TI - Stimulation of ATP secretion in the liver by therapeutic bile acids. AB - ATP receptors are ubiquitously expressed and are potential targets for the therapy of a number of disorders. However, delivery of ATP or other nucleotides to specific tissues is problematic, and no pharmacological means to stimulate the release of endogenous ATP has been described. We examined the effects of the bile acid ursodeoxycholic acid (UDCA) on ATP release into bile, since this bile acid is the only agent known to be of therapeutic benefit in secretory disorders of the liver, and since its mechanism of action is not established. Both UDCA and its taurine conjugate stimulated secretion of ATP by isolated rat hepatocytes, and produced measurable increases in ATP in bile of isolated rat liver. Perfusion of ATP into microdissected bile-duct segments induced Ca(2+) signalling in bile duct epithelia, while perfusion of bile acid did not. Thus UDCA may promote bile flow by inducing hepatocytes to release ATP into bile, which then stimulates fluid and electrolyte secretion by bile-duct epithelia downstream via changes in cytosolic Ca(2+). Moreover, these findings demonstrate the feasibility of using pharmacological means to induce secretion of endogenous ATP. Since the liver and other epithelial organs express luminal ATP receptors, these findings more generally suggest that a mechanism exists for pharmacological activation of this paracrine signalling pathway. This strategy may be useful for treatment of cystic fibrosis and other secretory disorders of the liver and other epithelial tissues. PMID- 11485546 TI - A large family of endosome-localized proteins related to sorting nexin 1. AB - Sorting nexin 1 (SNX1), a peripheral membrane protein, has previously been shown to regulate the cell-surface expression of the human epidermal growth factor receptor [Kurten, Cadena and Gill (1996) Science 272, 1008-1010]. Searches of human expressed sequence tag databases with SNX1 revealed eleven related human cDNA sequences, termed SNX2 to SNX12, eight of them novel. Analysis of SNX1 related sequences in the Saccharomyces cerevisiae genome clearly shows a greatly expanded SNX family in humans in comparison with yeast. On the basis of the predicted protein sequences, all members of this family of hydrophilic molecules contain a conserved 70-110-residue Phox homology (PX) domain, referred to as the SNX-PX domain. Within the SNX family, subgroups were identified on the basis of the sequence similarities of the SNX-PX domain and the overall domain structure of each protein. The members of one subgroup, which includes human SNX1, SNX2, SNX4, SNX5 and SNX6 and the yeast Vps5p and YJL036W, all contain coiled-coil regions within their large C-terminal domains and are found distributed in both membrane and cytosolic fractions, typical of hydrophilic peripheral membrane proteins. Localization of the human SNX1 subgroup members in HeLa cells transfected with the full-length cDNA species revealed a similar intracellular distribution that in all cases overlapped substantially with the early endosome marker, early endosome autoantigen 1. The intracellular localization of deletion mutants and fusions with green fluorescent protein showed that the C-terminal regions of SNX1 and SNX5 are responsible for their endosomal localization. On the basis of these results, the functions of these SNX molecules are likely to be unique to endosomes, mediated in part by interactions with SNX-specific C terminal sequences and membrane-associated determinants. PMID- 11485547 TI - Cartilage oligomeric matrix protein (COMP) and collagen IX are sensitive markers for the differentiation state of articular primary chondrocytes. AB - Primary chondrocytes dedifferentiate in serial monolayer with respect to their morphological and biosynthetic phenotype. They change from a round to a flattened fibroblast-like shape, and collagen I is secreted instead of the cartilage specific collagen II. We analysed in detail the time course of dedifferentiation of mature bovine articular chondrocytes in monolayer for up to 32 weeks. Assessment of RNA expression by reverse transcription-PCR led to the identification of two novel phenotypical markers, the cartilage oligomeric matrix protein (COMP) and collagen IX, which are down-regulated faster than the widely accepted marker, collagen II. The different kinetics of COMP and collagen expression suggest differential regulation at the level of transcription. Immunostaining and metabolic labelling experiments confirmed the switch in the collagen expression pattern and the rapid down-regulation of de novo synthesis of COMP and collagen IX. Culture of chondrocytes in a three-dimensional matrix is known to stabilize the chondrocytic phenotype. We maintained cells for up to 28 weeks in an alginate bead system, which prevented dedifferentiation and led to a stabilization of collagen and COMP expression. Immunohistochemical analysis of the alginate beads revealed a similar distribution of matrix proteins to that found in vivo. Chondrocytes were transferred after a variable length of monolayer culture into the alginate matrix and the potential for redifferentiation was investigated. The re-expression of COMP and collagen IX was differentially regulated. The expression of COMP was re-induced within days after transfer into the three-dimensional matrix, while the expression of collagen IX was irreversibly down-regulated. In summary, these results demonstrate that the potential for redifferentiation decreases with increasing length of monolayer culture and show that the alginate bead system represents an attractive in vitro model to study the chondrocyte de- and re-differentiation processes, as well as extracellular matrix assembly. PMID- 11485548 TI - Targeted overexpression of ornithine decarboxylase enhances beta-adrenergic agonist-induced cardiac hypertrophy. AB - These studies were designed to determine the consequences of constitutive overexpression of ornithine decarboxylase (ODC) in the heart. Induction of ODC is known to occur in response to agents that induce cardiac hypertrophy. However, it is not known whether high ODC levels are sufficient for the development of a hypertrophic phenotype. Transgenic mice were generated with cardiac-specific expression of a stable ODC protein using the alpha-myosin heavy-chain promoter. Founder lines with >1000-fold overexpression of ODC in the heart were established, resulting in a 50-fold overaccumulation of putrescine, 4-fold elevation in spermidine, a slight increase in spermine and accumulation of large amounts of cadaverine compared with littermate controls. Despite these significant alterations in polyamines, myocardial hypertrophy, as measured by ratio of heart to body weight, did not develop, although atrial natriuretic factor RNA was slightly elevated in transgenic ventricles. However, stimulation of beta-adrenergic signalling by isoproterenol resulted in severe hypertrophy and even death in ODC-overexpressing mice without further altering polyamine levels, compared with only a mild hypertrophy in littermates. When beta1-adrenergic stimulation was blocked by simultaneous treatment with isoproterenol and the beta1 antagonist atenolol, a significant, although reduced, hypertrophy was still present in the hearts of transgenic mice, suggesting that both beta1 and beta2 adrenergic receptors contribute to the hypertrophic phenotype. Therefore these mice provide a model to study the in vivo co-operativity between high ODC activity and activation of other pathways leading to hypertrophy in the heart. PMID- 11485549 TI - Steady-state localization of a medial-Golgi glycosyltransferase involves transit through the trans-Golgi network. AB - The steady-state localization of medial-Golgi enzymes is likely to involve retrograde transport pathways; however, the trafficking of these resident enzymes through the Golgi stack is unclear. To investigate if the medial-Golgi enzyme beta-1,2-N-acetylglucosaminyltransferase I (GlcNAc-TI) is transported to the late Golgi, a modified GlcNAc-TI bearing an N-glycan site on the C-terminus was constructed. The modified GlcNAc-TI was demonstrated to be functionally active in vivo, and was localized to the Golgi stack of transfected cells. In stable Chinese-hamster ovary (CHO) cell clones, the N-glycosylated GlcNAc-TI carried sialylated complex N-glycan chains. Pulse-chase studies showed that the majority of GlcNAc-TI was sialylated within 60 min of synthesis. Treatment of transfected CHO cells with Brefeldin A resulted in the glycosylated GlcNAc-TI bearing endo beta-N-acetylglucosaminidase H resistant chains; however, the sialylation of glycosylated GlcNAc-TI was dramatically reduced. These data imply that, in CHO cells, newly synthesized GlcNAc-TI is transported rapidly through the Golgi stack to the trans-Golgi network, suggesting that GlcNAc-TI continuously recycles from the late Golgi. Furthermore, this data suggests that retrograde transport pathways play an important role in establishing the asymmetric distribution of GlcNAc-TI within the Golgi stack. PMID- 11485550 TI - Overexpression of matrix metalloproteinase-2 mediates phenotypic transformation of lens epithelial cells. AB - Transforming growth factor-beta (TGF-beta) is known to be a causative factor in pathological fibrosis and the metastasis of cancer cells, through effects on molecules of the extracellular matrix (ECM). We evaluated the influence of TGF beta(1) on the gene expression of matrix metalloproteinase-2 (MMP-2) in lens epithelial cells (LECs). The results showed that TGF-beta(1) induced the expression of mRNA for MMP-2 in LECs. Subsequently, in order to examine the role of MMP-2, we overexpressed MMP-2 in LECs by stable transfection. The MMP-2 overexpressing LECs showed typical indicators of a myofibroblast-like cell phenotype, such as multiple layers of cells, elongated morphology, and expression of alpha-smooth muscle actin. We also showed that an MMP inhibitor blocked the TGF-beta(1)-induced morphological change in LECs. These results demonstrate that MMP-2 plays a role in the transformation of LECs, which has implications for the pathological fibrosis of these cells. PMID- 11485551 TI - The characterization of ligand-specific maize (Zea mays) profilin mutants. AB - Profilins are low-molecular-mass (12-15 kDa) cytosolic proteins that are major regulators of actin assembly in all eukaryotic cells. In general, profilins from evolutionarily diverse organisms share the ability to bind to G-actin, poly-(L proline) (PLP) and proline-rich proteins, and polyphosphoinositides. However, the functional importance of each of these interactions remains unclear and might differ between organisms. We investigated the importance of profilin's interaction with its various ligands in plant cells by characterizing four maize (Zea mays) profilin 5 (ZmPRO5) mutants that had single amino acid substitutions in the presumed sites of ligand interaction. Comparisons in vitro with wild-type ZmPRO5 showed that these mutations altered ligand association specifically. ZmPRO5-Y6F had a 3-fold increased affinity for PLP, ZmPRO5-Y6Q had a 5-fold decreased affinity for PLP, ZmPRO5-D8A had a 2-fold increased affinity for PtdIns(4,5)P(2) and ZmPRO5-K86A had a 35-fold decreased affinity for G-actin. When the profilins were microinjected into Tradescantia stamen hair cells, ZmPRO5 Y6F increased the rate of nuclear displacement in stamen hairs, whereas ZmPRO5 K86A decreased the rate. Mutants with a decreased affinity for PLP (ZmPRO5-Y6Q) or an enhanced affinity for PtdIns(4,5)P(2) (ZmPRO5-D8A) were not significantly different from wild-type ZmPRO5 in affecting nuclear position. These results indicate that plant profilin's association with G-actin is extremely important and further substantiate the simple model that profilin acts primarily as a G actin-sequestering protein in plant cells. Furthermore, interaction with proline rich binding partners might also contribute to regulating profilin's effect on actin assembly in plant cells. PMID- 11485552 TI - Multiple adaptive mechanisms affect asparagine synthetase substrate availability in asparaginase-resistant MOLT-4 human leukaemia cells. AB - Childhood acute lymphoblastic leukaemia is treated by combination chemotherapy with a number of drugs, almost always including the enzyme L-asparaginase (ASNase). Although the initial remission rate is quite high, relapse and associated drug resistance remain a problem. In vitro studies have demonstrated an adaptive increase in asparagine synthetase (AS) expression in ASNase-resistant cells, which is believed to permit ASNase-resistant human leukaemia cells to survive in vivo. The present results, obtained with ASNase-sensitive and resistant human MOLT-4 leukaemia cell lines, illustrate that several other adaptive processes occur to provide sufficient amounts of the AS substrates, aspartate and glutamine, required to support this increased enzymic activity. In both cell populations, aspartate is derived almost exclusively from intracellular sources, whereas the necessary glutamine arises from both intracellular and extracellular sources. Transport of glutamine into ASNase-resistant cells is significantly enhanced compared with the parental cells, whereas amino acid efflux (e.g. asparagine) is reduced. Most of the adaptive change for the amino acid transporters, Systems A, ASC and L, is rapidly (12 h) reversed following ASNase removal. The enzymic activity of glutamine synthetase is also enhanced in ASNase-resistant cells by a post-transcriptional mechanism. The results demonstrate that there are several sites of metabolic adaptation in ASNase treated leukaemia cells that serve to promote the replenishment of both glutamine and asparagine. PMID- 11485553 TI - Regulation of pyruvate dehydrogenase activity through phosphorylation at multiple sites. AB - The enzymic activity of the mammalian pyruvate dehydrogenase complex is regulated by the phosphorylation of three serine residues (sites 1, 2 and 3) located on the E1 component of the complex. Here we report that the four isoenzymes of protein kinase responsible for the phosphorylation and inactivation of pyruvate dehydrogenase (PDK1, PDK2, PDK3 and PDK4) differ in their abilities to phosphorylate the enzyme. PDK1 can phosphorylate all three sites, whereas PDK2, PDK3 and PDK4 each phosphorylate only site 1 and site 2. Although PDK2 phosphorylates site 1 and 2, it incorporates less phosphate in site 2 than PDK3 or PDK4. As a result, the amount of phosphate incorporated by each isoenzyme decreases in the order PDK1>PDK3>or=PDK4>PDK2. Significantly, binding of the coenzyme thiamin pyrophosphate to pyruvate dehydrogenase alters the rates and stoichiometries of phosphorylation of the individual sites. First, the rate of phosphorylation of site 1 by all isoenzymes of kinase is decreased. Secondly, thiamin pyrophosphate markedly decreases the amount of phosphate that PDK1 incorporates in sites 2 and 3 and that PDK2 incorporates in site 2. In contrast, the coenzyme does not significantly affect the total amount of phosphate incorporated in site 2 by PDK3 and PDK4, but instead decreases the rate of phosphorylation of this site. Furthermore, pyruvate dehydrogenase complex phosphorylated by the individual isoenzymes of kinase is reactivated at different rates by pyruvate dehydrogenase phosphatase. Both isoenzymes of phosphatase (PDP1 and PDP2) readily reactivate the complex phosphorylated by PDK2. When pyruvate dehydrogenase is phosphorylated by other isoenzymes, the rates of reactivation decrease in the order PDK4>or=PDK3>PDK1. Taken together, results reported here strongly suggest that the major determinants of the activity state of pyruvate dehydrogenase in mammalian tissues include the phosphorylation site specificity of isoenzymes of kinase in addition to the absolute amounts of kinase and phosphatase protein expressed in mitochondria. PMID- 11485554 TI - Apolipoprotein A-I stimulates the transport of intracellular cholesterol to cell surface cholesterol-rich domains (caveolae). AB - We have studied the effect of lipid-free human plasma apolipoprotein A-I (apoA-I) on the transport of newly synthesized cholesterol to cell-surface cholesterol rich domains, which in human skin fibroblasts are mainly represented by caveolae. Changes in transport of newly synthesized cholesterol were assessed after labelling cells with [(14)C]acetate at 15 degrees C and warming cells to permit the transfer of cholesterol, followed by the selective oxidation of cholesterol in cholesterol-rich domains (caveolae) in the plasma membrane before their partial purification. ApoA-I, but not BSA added in an equimolar concentration, enhanced the transport of cholesterol to the caveolae up to 5-fold in a dose- and time-dependent manner. The effect of apoA-I on cholesterol transport exceeded its effect on cholesterol efflux, resulting in an accumulation of intracellular cholesterol in caveolae. Methyl-beta-cyclodextrin, added at a concentration promoting cholesterol efflux to the same extent as apoA-I, also stimulated cholesterol trafficking, but was 3-fold less effective than apoA-I. Progesterone inhibited the transport of newly synthesized cholesterol to the caveolae. Treatment of cells with apoA-I stimulated the expression of caveolin, increasing the amount of caveolin protein and mRNA by approx. 2-fold. We conclude that apoA I induces the transport of intracellular cholesterol to cell-surface caveolae, possibly in part through the stimulation of caveolin expression. PMID- 11485555 TI - Assembly of protein kinase CK2: investigation of complex formation between catalytic and regulatory subunits using a zinc-finger-deficient mutant of CK2beta. AB - Protein kinase CK2 is a tetrameric enzyme comprised of two regulatory subunits (CK2beta) and two catalytic subunits (CK2alpha and/or CK2alpha'). The crystal structure of dimeric CK2beta demonstrated that a zinc finger mediates CK2beta dimerization, therefore we constructed a mutant in which cysteine residues 109 and 114 were mutated to serine. Our objectives were to examine the effects of disrupting the zinc finger of the regulatory CK2beta subunit on CK2 tetramer assembly. Examination of this zinc-finger-deficient mutant of CK2beta using a yeast two-hybrid assay demonstrates that the mutant fails to form CK2beta homodimers. In order to extend these studies, we co-transfected COS-7 cells with epitope-tagged constructs and performed co-immunoprecipitation assays. The results from these studies demonstrate that the mutant fails to form CK2beta homodimers and fails to interact with catalytic CK2 subunits. Furthermore, we demonstrate that the mutant CK2beta is not appreciably phosphorylated in cells. Using in vitro binding assays, we demonstrated that the mutant CK2beta protein fails to interact with glutathione S-transferase-CK2alpha'. Finally, we demonstrate that the mutant is translated at an equivalent rate to wild-type CK2beta, but is degraded much more rapidly. Overall, our results are consistent with the model that beta-beta dimerization precedes incorporation of catalytic subunits into tetrameric CK2 complexes, and that beta-beta dimerization is a prerequisite for the stable incorporation of catalytic subunits into CK2 complexes. PMID- 11485556 TI - A maize insulin-like growth factor signals to a transduction pathway that regulates protein synthesis in maize. AB - Insulin and insulin-like growth factors (IGFs) are well-characterized regulators in higher eukaryotic cells that control biological processes such as cell growth and survival, and selective translation of mRNAs. This research presents the purification of a 20 kDa protein, isolated from maize tissue, with IGF activity. The protein was purified from 48 h-germinated maize embryonic axes by G-50 Sephadex fractionation followed by affinity chromatography through a bovine insulin antibody-Sepharose column. This protein proved to significantly speed up maize germination and seedling growth. At the molecular level, Zea mays IGF (ZmIGF) enhanced phosphorylation of S6 ribosomal protein (rp) on the 40 S ribosomal subunit, in a similar way as observed when bovine insulin is applied to maize axes during germination. Rapamycin, a specific inhibitor of the insulin stimulated signal transduction pathway, prevented S6 rp phosphorylation in maize axes. Moreover, ZmIGF stimulated [(35)S]methionine incorporation into rps, above the level of overall cytoplasmic proteins. Either incubation with anti-insulin antibody, heat treatment (60 degrees C) or trypsin digestion abolished this ZmIGF effect. It is proposed that ZmIGF is an endogenous maize growth factor that regulates the synthesis of specific proteins through a pathway similar to that of insulin or IGFs in animal tissues. PMID- 11485557 TI - Functional and structural roles of the glutathione-binding residues in maize (Zea mays) glutathione S-transferase I. AB - The isoenzyme glutathione S-transferase (GST) I from maize (Zea mays) was cloned and expressed in Escherichia coli, and its catalytic mechanism was investigated by site-directed mutagenesis and dynamic studies. The results showed that the enzyme promotes proton dissociation from the GSH thiol and creates a thiolate anion with high nucleophilic reactivity by lowering the pK(a) of the thiol from 8.7 to 6.2. Steady-state kinetics fit well to a rapid equilibrium, random sequential Bi Bi mechanism, with intrasubunit modulation between the GSH binding site (G-site) and the electrophile binding site (H-site). The rate-limiting step of the reaction is viscosity-dependent, and thermodynamic data suggest that product release is rate-limiting. Five residues of GST I (Ser(11), His(40), Lys(41), Gln(53) and Ser(67)), which are located in the G-site, were individually replaced with alanine and their structural and functional roles in the 1-chloro 2,4-dinitrobenzene (CDNB) conjugation reaction were investigated. On the basis of steady-state kinetics, difference spectroscopy and limited proteolysis studies it is concluded that these residues: (1) contribute to the affinity of the G-site for GSH, as they are involved in side-chain interaction with GSH; (2) influence GSH thiol ionization, and thus its reactivity; (3) participate in k(cat) regulation by affecting the rate-limiting step of the reaction; and (4) in the cases of His(40), Lys(41) and Gln(53) play an important role in the structural integrity of, and probably in the flexibility of, the highly mobile short 3(10) helical segment of alpha-helix 2 (residues 35-46), as shown by limited proteolysis experiments. These structural perturbations are probably transmitted to the H-site through changes in Phe(35) conformation. This accounts for the modulation of K(CDNB)(m) by His(40), Lys(41) and Gln(53), and also for the intrasubunit communication between the G- and H-sites. Computer simulations using CONCOORD were applied to maize GST I monomer and dimer structures, each with bound lactoylglutathione, and the results were analysed by the essential dynamics technique. Differences in dynamics were found between the monomer and the dimer simulations showing the importance of using the whole structure in dynamic analysis. The results obtained confirm that the short 3(10)-helical segment of alpha-helix 2 (residues 35-46) undergoes the most significant structural rearrangements. These rearrangements are discussed in terms of enzyme catalytic mechanism. PMID- 11485558 TI - Structural and kinetic modifications of aldose reductase by S-nitrosothiols. AB - Modification of aldose reductase (AR) by the nitrosothiols S-nitroso-N-acetyl penicillamine (SNAP) and N-(beta-glucopyranosyl)-N(2)-acetyl-S nitrosopenicillamide (glyco-SNAP) resulted in a 3-7-fold increase in its k(cat) and a 25-40-fold increase in its K(m) for glyceraldehyde. In comparison with the native protein, the modified enzyme was less sensitive to inhibition by sorbinil and was not activated by SO(2-)(4) anions. The active-site residue, Cys-298, was identified as the main site of modification, because the site-directed mutant in which Cys-298 was replaced by serine was insensitive to glyco-SNAP. The extent of modification was not affected by P(i) or O(2), indicating that it was not due to spontaneous release of nitric oxide (NO) by the nitrosothiols. Electrospray ionization MS revealed that the modification reaction proceeds via the formation of an N-hydroxysulphenamide-like adduct between glyco-SNAP and AR. In time, the adduct dissociates into either nitrosated AR (AR-NO) or a mixed disulphide between AR and glyco-N-acetylpenicillamine (AR-S-S-X). Removal of the mixed disulphide form of the protein by lectin-column chromatography enriched the preparation in the high-K(m)-high-k(cat) form of the enzyme, suggesting that the kinetic changes are due to the formation of AR-NO, and that the AR-S-S-X form of the enzyme is catalytically inactive. Modification of AR by the non-thiol NO donor diethylamine NONOate (DEANO) increased enzyme activity and resulted in the formation of AR-NO. However, no adducts between AR and DEANO were formed. These results show that nitrosothiols cause multiple structural and functional changes in AR. Our observations also suggest the general possibility that transnitrosation reactions can generate both nitrosated and thiolated products, leading to non-unique changes in protein structure and function. PMID- 11485560 TI - Genomic distribution of three promoters of the bovine gene encoding acetyl-CoA carboxylase alpha and evidence that the nutritionally regulated promoter I contains a repressive element different from that in rat. AB - The enzyme acetyl-CoA carboxylase alpha (ACC-alpha) is rate-limiting for the synthesis of long-chain fatty acids de novo. As a first characterization of the bovine gene encoding this enzyme, we established the entire bovine ACC-alpha cDNA sequence (7041 bp) and used experiments with 5' rapid amplification of cDNA ends to determine the heterogeneous composition of 5' untranslated regions, as expressed from three different promoters (PI, PII and PIII). The individual locations of these promoters have been defined within an area comprising 35 kbp on Bos taurus chromosome 19 ('BTA19'), together with the segmentation of the first 14 exons. Primer extension analyses reveal that the nutritionally regulated PI initiates transcription from at least four sites. PI transcripts are much more abundant in adipose and mammary-gland tissues than in liver or lung. A 2.6 kb promoter fragment drives the expression of reporter genes only weakly in different model cells, irrespective of stimulation with insulin or dexamethasone. Thus bovine PI is basically repressed, like its analogue from rat. Finely graded deletions of PI map two separate elements, which have to be present together in cis to repress bovine PI. The distal component resides within a well-preserved Art2 retroposon element. Thus sequence, structure and evolutionary origin of the main repressor of PI in bovines are entirely different from its functional counterpart in rat, which had been identified as a (CA)(28) microsatellite. We show that, in different mammalian species, unrelated genome segments of different origins have been recruited to express as functionally homologous PI the ancient and otherwise highly conserved ACC-alpha-encoding gene. PMID- 11485559 TI - Selective induction of phospholipase D1 in pathogen-activated human monocytes. AB - Phospholipase D (PLD) activation is part of the complex signalling cascade induced during phagocyte activation. Two PLD isoforms have been cloned, but their role in phagocyte functions is still poorly defined. We report that resting fresh circulating human monocytes expressed PLD1. PLD1 protein expression was rapidly down-regulated during cell culture. Lipopolysaccharide and pathogen-derived agonists (Candida albicans, arabinoside-terminated lipoarabinomannan and Gram positive bacteria, but not mannose-capped lipoarabinomannan or double-stranded RNA) strongly induced PLD1 expression at both the mRNA and protein levels. Pro inflammatory cytokines [interleukin (IL)-1beta and tumour necrosis factor alpha] had only a weak effect, whereas immune cytokines (IL-6 and interferon gamma), anti-inflammatory cytokines (IL-13 and IL-10) and chemoattractants (fMet-Leu-Phe and macrophage chemoattractant protein 1) were inactive. None of the agonists tested induced significant changes in the basal expression of PLD2 mRNA. Consistent with PLD1 up-regulation was the observation that PLD enzymic activity was higher in monocytes treated with active-pathogen-derived agonists than in control cells, when stimulated with PMA or with chemotactic agonists (fMet-Leu Phe and C5a). Thus PLD2 seems to be a constitutive enzyme in circulating monocytes. Conversely, PLD1 is an inducible protein, rapidly regulated during culture conditions and selectively induced during cell activation. Therefore PLD1 might have a relevant role in immune responses against pathogens and in chronic inflammation. PMID- 11485561 TI - Polyamine analogues inhibit the ubiquitination of spermidine/spermine N1 acetyltransferase and prevent its targeting to the proteasome for degradation. AB - Spermidine/spermine N(1)-acetyltransferase (SSAT), a key enzyme in mammalian polyamine catabolism, undergoes rapid turnover (half-life approx. 30 min) and is highly inducible in response to polyamine analogues such as bis(ethyl)spermine (BE-3-4-3), which greatly stabilize the enzyme. Rapid degradation of SSAT in reticulocyte lysates was preceded by formation of a ladder of ubiquitinated forms, and required the production of high-molecular-mass complexes with ubiquitin (HMM-SSAT-Ubs). Mutation of all 11 lysines in SSAT separately to arginine demonstrated that no single lysine residue is critical for its degradation in vitro, but mutant K87R had a significantly longer half-life, suggesting that lysine-87 may be the preferred site for ubiquitination. Mutations at the C-terminus of SSAT, such as E171Q, resulted in marked stabilization of the protein, due to the lack of formation of the HMM-SSAT-Ubs. Addition of BE-3-4-3 prevented the accumulation of ubiquitin conjugates and the proteasomal degradation of wild-type SSAT. These results indicate that conformational changes brought about by the binding of polyamine analogues prevent the efficient polyubiquitination of SSAT, leading to a major increase in the amount of SSAT protein, and that alteration of the C-terminal end of the protein has a similar effect in preventing the productive interaction with an E2 or E3 component of the ubiquitin pathway. PMID- 11485562 TI - Calcium binding and translocation by the voltage-dependent anion channel: a possible regulatory mechanism in mitochondrial function. AB - Mitochondria play a central role in energy metabolism, Ca(2+) signalling, aging and cell death. To control cytosolic or mitochondrial Ca(2+) concentration, mitochondria possess several Ca(2+)-transport systems across the inner membrane. However, the pathway for Ca(2+) crossing the outer membrane has not been directly addressed. We report that purified voltage-dependent anion channel (VDAC) reconstituted into lipid bilayers or liposomes is highly permeable to Ca(2+). VDAC contains Ca(2+)-binding sites that bind Ruthenium Red (RuR), La(3+) and that RuR completely closed VDACs in single or multichannel experiments. Energized, freshly prepared mitochondria accumulate Ca(2+) (500-700 nmol/mg of protein), and subsequently released it. The release of Ca(2+) is accompanied by cyclosporin A inhibited swelling, suggesting activation of permeability transition pore (PTP). RuR and ruthenium amine binuclear complex, when added to mitochondria after Ca(2+) accumulation has reached a maximal level and before PTP is activated, prevented the release of Ca(2+) and the accompanied mitochondrial swelling. RuR also prevented PTP opening promoted by atractyloside, an adenine nucleotide translocase inhibitor. These results suggest that VDAC, located in the mitochondrial outer membrane, controls Ca(2+) transport into and from the mitochondria, and that the inhibition of Ca(2+) uptake by RuR and La(3+) may result from their interaction with VDAC Ca(2+)-binding sites. Inhibition of PTP opening or assembly by RuR and ruthenium amine binuclear complex suggest the involvement of VDAC in PTP activity and/or regulation. The permeability of VDAC to Ca(2+) and its binding of Ca(2+), suggest that VDAC has a role in regulation of the mitochondrial Ca(2+) homoeostasis. PMID- 11485563 TI - Elongated dermatan sulphate in post-inflammatory healing skin distributes among collagen fibrils separated by enlarged interfibrillar gaps. AB - It has been reported that the disaccharide composition of dermatan sulphate shows transient changes after epicutaneous application of the hapten 2,4 dinitrofluorobenzene to mouse skin, and that these changes are most conspicuous in healing skin on day 15 after chemical insult [Kuwaba, Nomura, Irie and Koyama (1999) J. Dermatol. Sci. 19, 23-30]. In the present study it was found that the molecular size of dermatan sulphate was increased on day 15 after hapten application. The molecular size of decorin increased in healing skin, whereas the size of dermatan-sulphate-depleted core protein did not increase. The length and localization of decorin dermatan sulphate were investigated by electron microscopy. Dermatan sulphate filaments oriented orthogonally to collagen fibrils were longer in healing skin than in control skin. In control skin, dermatan sulphate filaments were found among tightly packed collagen fibrils. In contrast, the interfibrillar gaps between each collagen fibril were enlarged in healing skin; elongated dermatan sulphate filaments extended from the surface of collagen fibrils across the enlarged gap. These results suggest that the increase in molecular size of decorin dermatan sulphate is important in organizing collagen fibrils separated by enlarged interfibrillar gaps in healing skin. PMID- 11485564 TI - A fluorescence temperature-jump study of conformational transitions in myosin subfragment 1. AB - The tryptophan fluorescence of unmodified myosin subfragment 1 (S1) from rabbit and chicken skeletal muscle with various nucleotides and phosphate analogues bound was measured after rapid temperature jumps. The fluorescence decreased during the temperature rise. Under some conditions, this decrease was followed by an increase, reflecting structural transitions within the protein. With adenosine 5'-[beta,gamma-imido]triphosphate (p[NH]ppA) or with ADP and BeF(x) bound, this rise was very rapid (reciprocal time constant approx. 2000 s(-1)) and varied only slightly with starting temperature, suggesting that, with these ligands, two different protein conformations were present in rapid equilibrium over a large temperature range. In the presence of ATP, the transient included several relaxation processes. Overall, the results suggest that complexes of S1 with ATP or with a number of other ligands exist as a mixture of two forms in temperature dependent equilibrium. The results throw light on the finding of different forms of S1 in recent crystallographic studies and indicate a surprising lack of strong coupling between myosin's structural state and the nature of the nucleotide bound. PMID- 11485566 TI - Roles of the juxtamembrane and extracellular domains of angiotensin-converting enzyme in ectodomain shedding. AB - Angiotensin-converting enzyme (ACE) is one of a growing number of integral membrane proteins that is shed from the cell surface through proteolytic cleavage by a secretase. To investigate the requirements for ectodomain shedding, we replaced the glycosylphosphatidylinositol addition sequence in membrane dipeptidase (MDP) - a membrane protein that is not shed - with the juxtamembrane stalk, transmembrane (TM) and cytosolic domains of ACE. The resulting construct, MDP-STM(ACE), was targeted to the cell surface in a glycosylated and enzymically active form, and was shed into the medium. The site of cleavage in MDP-STM(ACE) was identified by MS as the Arg(374)-Ser(375) bond, corresponding to the Arg(1203)-Ser(1204) secretase cleavage site in somatic ACE. The release of MDP STM(ACE) and ACE from the cells was inhibited in an identical manner by batimastat and two other hydroxamic acid-based zinc metallosecretase inhibitors. In contrast, a construct lacking the juxtamembrane stalk, MDP-TM(ACE), although expressed at the cell surface in an enzymically active form, was not shed, implying that the juxtamembrane stalk is the critical determinant of shedding. However, an additional construct, ACEDeltaC, in which the N-terminal domain of somatic ACE was fused to the stalk, TM and cytosolic domains, was also not shed, despite the presence of a cleavable stalk, implying that in contrast with the C terminal domain, the N-terminal domain lacks a signal required for shedding. These data are discussed in the context of two classes of secretases that differ in their requirements for recognition of substrate proteins. PMID- 11485565 TI - Spot 14 protein interacts and co-operates with chicken ovalbumin upstream promoter-transcription factor 1 in the transcription of the L-type pyruvate kinase gene through a specificity protein 1 (Sp1) binding site. AB - In hepatocytes, the amount of the Spot 14 (S14) protein is closely related to the full expression of enzymes involved in the glycolytic and lipogenic pathways. In the present study we address the role played by this protein in the control of transcription of the L-type pyruvate kinase (L-PK) gene in primary hepatocytes. We show that human S14, which by itself does not bind to the L-PK promoter, physically interacts with the human chicken ovalbumin upstream promoter transcription factor 1 (COUP-TF1) and induces the switch of this factor from a repressor to an activator. However, the enhancing activity of S14 and COUP-TF1 depends on the presence of a proximal GC-rich box (the L0 element) that specifically binds nuclear proteins from the livers of rats fed a glucose-rich diet. Moreover, the L0 element, which strongly binds dephosphorylated specificity protein 1 (Sp1), loses all affinity when this factor is phosphorylated by cAMP dependent protein kinase. Mutations that affect binding of Sp1 and nuclear proteins to the L0 box also decrease basal transcription and impair glucose responsiveness of the promoter. These results therefore shed light on the mechanism by which the S14 protein, whose concentration rapidly rises after glucose intake, contributes to the full activity of the L-PK promoter. PMID- 11485567 TI - Protein kinase C delta and eta isoenzymes control the shedding of the interleukin 6 receptor alpha in myeloma cells. AB - The soluble interleukin 6 receptor alpha is an agonistic molecule of interleukin 6 (IL-6) and is important in the biology of multiple myeloma. More precisely, it potentiates the deleterious effects of IL-6 during tumour progression, facilitating angiogenesis and bone resorption. Because the mechanisms involved in the shedding of the interleukin 6 receptor alpha (IL-6Ralpha) in multiple myeloma are not known, we have investigated them in the XG-6 human myeloma cell line. Here we provide evidence that PMA-induced IL-6Ralpha shedding is controlled by a metalloproteinase and by protein kinase C (PKC) isoenzymes that do not require Ca(2+) for their activation. We show that XG-6 cells express PKC-delta, -eta and zeta isoenzymes. However, after stimulation with PMA, only PKC-delta and PKC-eta are activated, as shown by their translocation to the membrane. Treatment with PMA induces an increase in PKC-delta phosphorylation in its active loop. In addition, by using rottlerin, a specific inhibitor of PKC-delta, we demonstrate that PKC-delta is involved in the PMA-induced shedding of IL-6Ralpha. With the use of UO126, a specific inhibitor of the mitogen-activated protein kinase (MAPK) pathway, we show that the PMA-induced IL-6Ralpha shedding is mediated in part by the MAPK pathway. Finally, whereas GF109203X, a general PKC inhibitor, inhibits the activation of ERK1/2 (extracellular signal-regulated protein kinase 1/2), rottlerin has no inhibitory effect, indicating that the Ras/MAPK activation is PKC-dependent but PKC-delta-independent. Taken together, these results suggest that the PMA-induced shedding of IL-6Ralpha is mediated by a PKC isoenzyme network. PMID- 11485568 TI - Kinetics of CO binding to the haem domain of murine inducible nitric oxide synthase: differential effects of haem domain ligands. AB - The binding of CO to the murine inducible nitric oxide synthase (iNOS) oxygenase domain has been studied by laser flash photolysis. The effect of the (6R)-5,6,7,8 tetrahydro-L-biopterin (BH(4)) cofactor L-arginine and several Type I L-arginine analogues/ligands on the rates of CO rebinding has been evaluated. The presence of BH(4) in the iNOS active site has little effect on the rebinding of protein caged haem-CO pairs (geminate recombination), but decreases the bimolecular association rates 2-fold. Addition of L-arginine to the BH(4)-bound complex completely abolishes geminate recombination and results in a further 80-fold decrease in the overall rate of bimolecular association. Three of the Type I ligands, S-ethylisothiourea, L-canavanine and 2,5-lutidine, displaced the CO from the haem iron upon addition to the iNOS oxygenase domain. The Type I ligands significantly decreased the rate of bimolecular binding of CO to the haem iron after photolysis. Most of these ligands also completely abolished geminate recombination. These results are consistent with a relatively open distal pocket that allows CO to bind unhindered in the active site of murine iNOS in the absence of L-arginine or BH(4). In the presence of BH(4) and L-arginine, however, the enzyme adopts a more closed structure that can greatly reduce ligand access to the haem iron. These observations are discussed in terms of the known structure of iNOS haem domain and solution studies of ligand binding in iNOS and neuronal NOS isoenzymes. PMID- 11485569 TI - Differential effects of glycosphingolipids on the detergent-insolubility of the glycosylphosphatidylinositol-anchored membrane dipeptidase. AB - The insolubility of glycosylphosphatidylinositol (GPI)-anchored proteins in certain detergents appears to be an intrinsic property of their association with sphingolipids and cholesterol in lipid rafts. We show that the GPI-anchored protein membrane dipeptidase is localized in detergent-insoluble lipid rafts isolated from porcine kidney microvillar membranes, and that these rafts, which lack caveolin, are enriched not only in sphingomyelin and cholesterol, but also in the glycosphingolipid lactosylceramide (LacCer). Dipeptidase purified from porcine kidney was reconstituted into artificial liposomes in order to investigate the relationship between glycosphingolipids and GPI-anchored protein detergent-insolubility. Dipeptidase was insoluble in liposomes containing extremely low concentrations of LacCer. In contrast, identical concentrations of glucosylceramide or galactosylceramide failed to promote significant detergent insolubility. Cholesterol was shown to enhance the detergent-insoluble effect of LacCer. GC-MS analysis revealed dramatic differences between the fatty acyl compositions of LacCer and those of the other glycosphingolipids. However, despite these differences, we show that the unusually marked effect of LacCer to promote the detergent-insolubility of dipeptidase cannot be singularly attributed to the fatty acyl composition of this glycosphingolipid molecule. Instead, we suggest that the ability of LacCer to confer detergent-insolubility on this GPI anchored protein is dependent on the structure of the lipid molecule in its entirety, and that this glycosphingolipid may have an important role to play in the stabilization of lipid rafts, particularly the caveolin-free glycosphingolipid signalling domains. PMID- 11485570 TI - Endoplasmic reticulum Ca2+-ATPase pump is up-regulated in calcium-transporting dental enamel cells: a non-housekeeping role for SERCA2b. AB - Dental enamel-forming cells face a major challenge to avoid the cytotoxic effects of excess calcium. We have characterized sarcoplasmic/endoplasmic reticulum calcium-ATPase pumps (SERCA) in rat enamel cells to address the proposal that non mitochondrial calcium stores play a dominant role in transcellular calcium transport. A single major isoform, SERCA2b, was detected during the protein secretory and calcium-transport stages of enamel formation using reverse transcriptase PCR, cDNA cloning, Northern analysis and immunoblotting. Most importantly, SERCA2b exhibited a specific 3-fold up-regulation to high expression levels during calcium transport, as determined by quantitative immunoblotting and ATPase assays. Sensitivity of the calcium-dependent ATPase to thapsigargin and three other SERCA inhibitors was characterized. These findings indicate that enamel cells are well-equipped to sequester calcium in endoplasmic reticulum stores and so protect against calcium toxicity, associate SERCA with transcellular calcium transport for the first time, and establish SERCA2b as a molecular and pharmacological target for future investigations of calcium transcytosis. The observed physiological regulation in enamel cells contradicts the widespread perception that SERCA2b is restricted to general housekeeping duties. PMID- 11485571 TI - Structure and expression of human mitochondrial adenylate kinase targeted to the mitochondrial matrix. AB - The previously isolated cDNA encoding human adenylate kinase (AK) isozyme 3 was recently renamed AK4. Consequently, human AK3 cDNA remains to be identified and we have little information about the functional relationship between human AK3 and AK4. In pursuit of the physiological roles of both the AK3 and AK4 proteins, we first isolated an authentic human AK3 cDNA and compared their expression. Nucleotide sequencing revealed that the cDNA encoded a 227-amino-acid protein, with a deduced molecular mass of 25.6 kDa, that shares greater homology with the AK3 cDNAs isolated from bovine and rat than that from human. We named the isolated cDNA AK3. Northern-blot analysis revealed that AK3 mRNA was present in all tissues examined, and was highly expressed in heart, skeletal muscle and liver, moderately expressed in pancreas and kidney, and weakly expressed in placenta, brain and lung. On the other hand, we found that human AK4 mRNA was highly expressed in kidney, moderately expressed in heart and liver and weakly expressed in brain. Western-blot analysis demonstrated expression profiles of AK3 and AK4 that were similar to their mRNA expression patterns in each tissue. Over expression of AK3, but not AK4, in both Escherichia coli CV2, a temperature sensitive AK mutant, and a human embryonic kidney-derived cell line, HEK-293, not only produced significant GTP:AMP phosphotransferase (AK3) activity, but also complemented the CV2 cells at 42 degrees C. Subcellular and submitochondrial fractionation analysis demonstrated that both AK3 and AK4 are localized in the mitochondrial matrix. PMID- 11485572 TI - Substrates and products of eosinophil peroxidase. AB - Eosinophil peroxidase has been implicated in promoting oxidative tissue damage in a variety of inflammatory conditions, including asthma. It uses H(2)O(2) to oxidize chloride, bromide and thiocyanate to their respective hypohalous acids. The aim of this study was to establish which oxidants eosinophil peroxidase produces under physiological conditions. By measuring rates of H(2)O(2) utilization by the enzyme at neutral pH, we determined the catalytic rate constants for bromide and thiocyanate as 248 and 223 s(-1) and the Michaelis constants as 0.5 and 0.15 mM respectively. On the basis of these values thiocyanate is preferred 2.8-fold over bromide as a substrate for eosinophil peroxidase. Eosinophil peroxidase catalysed substantive oxidation of chloride only below pH 6.5. We found that when eosinophil peroxidase or myeloperoxidase oxidized thiocyanate, another product besides hypothiocyanite was formed; it also converted methionine into methionine sulphoxide. During the oxidation of thiocyanate, the peroxidases were present as their compound II forms. Compound II did not form when GSH was included to scavenge hypothiocyanite. We propose that the unidentified oxidant was derived from a radical species produced by the one electron oxidation of hypothiocyanite. We conclude that at plasma concentrations of bromide (20-120 microM) and thiocyanate (20-100 microM), hypobromous acid and oxidation products of thiocyanate are produced by eosinophil peroxidase. Hypochlorous acid is likely to be produced only when substrates preferred over chloride are depleted. Thiocyanate should be considered to augment peroxidase mediated toxicity because these enzymes can convert relatively benign hypothiocyanite into a stronger oxidant. PMID- 11485573 TI - Second generation antitumour human RNase: significance of its structural and functional features for the mechanism of antitumour action. AB - A second generation mutant of dimeric human pancreas RNase (HHP2-RNase), was obtained by a single residue mutation (Glu(111)-->Gly) of the previously described dimeric human pancreas RNase variant (HHP-RNase). HHP2-RNase was found to be a highly specific antitumour agent, with an enhanced cytotoxic activity compared with HHP-RNase. The structural and functional requisites of the antitumour action of HHP2-RNase were investigated and compared with those of other dimeric antitumour RNases. The stability of the dimeric structure, i.e. the resistance of human dimeric RNase variants to reductive cleavage of the two intersubunit disulphide bonds that bridge the subunits, was determined to be an essential feature of antitumour dimeric RNases. The stability of the dimeric structure is in turn responsible for the resistance to inhibition by the cytosolic RNase inhibitor (cRI). Both the stability of the dimeric structure and the resistance to cRI inhibition appeared to be highly enhanced by an RNase substrate. This suggests a possible role for RNA in the amplification of the antitumour potential of dimeric RNases. PMID- 11485575 TI - Purification and characterization of a glutathione S-transferase Omega in pig: evidence for two distinct organ-specific transcripts. AB - A cytosolic glutathione S-transferase (GST, EC 2.5.1.18) from the recently characterized Omega class [GSTO; Board et al. 2000, J. Biol. Chem. 275, 24798 24806] has been identified in pig organs. It was found widely distributed in the different tissues investigated and especially abundant in liver and muscle. The hepatic enzyme has been purified to homogeneity by using its selective affinity for S-hexylglutathione over GSH, thus providing a simple method to isolate mammalian GSTO. The dimeric protein has a subunit molecular mass of 27328 Da as measured by electrospray ionization MS. Internal peptide sequencing and complete cDNA sequencing revealed strong similarities with its human recombinant orthologue and two rodent GST-like proteins with the ability to catalyse the GSH dependent reduction of dehydroascorbate. Additional similarities, including the presence of a specific N-terminal extension and of immunological cross reactivity, support the results. Moreover, this gene encoding GSTO generates two organ-specific transcripts, suggesting transcriptional mechanisms with a significance that is as yet uncharacterized. PMID- 11485574 TI - Palmitoylethanolamide inhibits the expression of fatty acid amide hydrolase and enhances the anti-proliferative effect of anandamide in human breast cancer cells. AB - Palmitoylethanolamide (PEA) has been shown to act in synergy with anandamide (arachidonoylethanolamide; AEA), an endogenous agonist of cannabinoid receptor type 1 (CB(1)). This synergistic effect was reduced by the CB(2) cannabinoid receptor antagonist SR144528, although PEA does not activate either CB(1) or CB(2) receptors. Here we show that PEA potently enhances the anti-proliferative effects of AEA on human breast cancer cells (HBCCs), in part by inhibiting the expression of fatty acid amide hydrolase (FAAH), the major enzyme catalysing AEA degradation. PEA (1-10 microM) enhanced in a dose-related manner the inhibitory effect of AEA on both basal and nerve growth factor (NGF)-induced HBCC proliferation, without inducing any cytostatic effect by itself. PEA (5 microM) decreased the IC(50) values for AEA inhibitory effects by 3-6-fold. This effect was not blocked by the CB(2) receptor antagonist SR144528, and was not mimicked by a selective agonist of CB(2) receptors. PEA enhanced AEA-evoked inhibition of the expression of NGF Trk receptors, which underlies the anti-proliferative effect of the endocannabinoid on NGF-stimulated MCF-7 cells. The effect of PEA was due in part to inhibition of AEA degradation, since treatment of MCF-7 cells with 5 microM PEA caused a approximately 30-40% down-regulation of FAAH expression and activity. However, PEA also enhanced the cytostatic effect of the cannabinoid receptor agonist HU-210, although less potently than with AEA. PEA did not modify the affinity of ligands for CB(1) or CB(2) receptors, and neither did it alter the CB(1)/CB(2)-mediated inhibitory effect of AEA on adenylate cyclase type V, nor the expression of CB(1) and CB(2) receptors in MCF-7 cells. We suggest that long-term PEA treatment of cells may positively affect the pharmacological activity of AEA, in part by inhibiting FAAH expression. PMID- 11485576 TI - Matrix-assisted in vitro refolding of Pseudomonas aeruginosa class II polyhydroxyalkanoate synthase from inclusion bodies produced in recombinant Escherichia coli. AB - In order to facilitate the large-scale preparation of active class II polyhydroxyalkanoate (PHA) synthase, we constructed a vector pT7-7 derivative that contains a modified phaC1 gene encoding a PHA synthase from Pseudomonas aeruginosa possessing six N-terminally fused histidine residues. Overexpression of this phaC1 gene under control of the strong O10 promoter was achieved in Escherichia coli BL21(DE3). The fusion protein was deposited as inactive inclusion bodies in recombinant E. coli, and contributed approx. 30% of total protein. The inclusion bodies were purified by selective solubilization, resulting in approx. 70-80% pure PHA synthase, then dissolved and denatured by 6 M guanidine hydrochloride. The denatured PHA synthase was reversibly immobilized on a Ni(2+)-nitrilotriacetate-agarose matrix. The matrix-bound fusion protein was refolded by gradual removal of the chaotropic reagent. This procedure avoided the aggregation of folding intermediates which often decreases the efficiency of refolding experiments. Finally, the refolded fusion protein was eluted with imidazole. The purified and refolded PHA synthase protein showed a specific enzyme activity of 10.8 m-units/mg employing (R/S)-3-hydroxydecanoyl-CoA as substrate, which corresponds to 27% of the maximum specific activity of the native enzyme. The refolding of the enzyme was confirmed by CD spectroscopy. Deconvolution of the spectrum resulted in the following secondary structure prediction: 10% alpha-helix, 50% beta-sheet and 40% random coil. Gel filtration chromatography indicated an apparent molecular mass of 69 kDa for the refolded PHA synthase. However, light-scattering analysis of a 10-fold concentrated sample indicated a molecular mass of 128 kDa. These data suggest that the class II PHA synthase is present in an equilibrium of monomer and dimer. PMID- 11485577 TI - Mutation of yeast Eug1p CXXS active sites to CXXC results in a dramatic increase in protein disulphide isomerase activity. AB - Protein disulphide isomerase (PDI) is an essential protein which is localized to the endoplasmic reticulum of eukaryotic cells. It catalyses the formation and isomerization of disulphide bonds during the folding of secretory proteins. PDI is composed of domains with structural homology to thioredoxin and with CXXC catalytic motifs. EUG1 encodes a yeast protein, Eug1p, that is highly homologous to PDI. However, Eug1p contains CXXS motifs instead of CXXC. In the current model for PDI function both cysteines in this motif are required for PDI-catalysed oxidase activity. To gain more insight into the biochemical properties of this unusual variant of PDI we have purified and characterized the protein. We have furthermore generated a number of mutant forms of Eug1p in which either or both of the active sites have been mutated to a CXXC sequence. To determine the catalytic capacity of the wild-type and mutant forms we assayed activity in oxidative refolding of reduced and denatured procarboxypeptidase Y as well as refolding of bovine pancreatic trypsin inhibitor. The wild-type protein showed very little activity, not only in oxidative refolding but also in assays where only isomerase activity was required. This was surprising, in particular since mutant forms of Eug1p containing a CXXC motif displayed activity close to that of genuine PDI. These results lead us to propose that general disulphide isomerization is not the main function of Eug1p in vivo. PMID- 11485579 TI - Ear deformity in children following high ear-piercing: current practice, consent issues and legislation. AB - In this presentation we examine the practice of high ear-piercing in children, the issue of informed consent and current legislation. We sampled current practice and consent policy by visiting nine establishments in Sheffield providing this service. There were two high street department stores, two fashion accessory outlets and five body-piercing studios. Enquiries were made as to the technique used, knowledge of complications, customer counselling and consent policy. A photograph of an ear with a cosmetic deformity following high ear piercing was shown and awareness of this possible outcome was noted. Two ear piercing techniques were identified, either a spring-loaded gun firing a blunt stud or the use of a body-piercing needle. The fashion accessory outlets were prepared to pierce any part of the ear using a spring-loaded gun in children under 16 years of age. There was a general lack of knowledge about possible serious complications. Two of the body piercers would not perform high ear piercing on clients under the age of 16 years. The body piercers use a disposable needle and were of the opinion that using a spring-loaded gun shatters the cartilage and increases the risk of infection. The best technique is open to debate and it may be that the perceived unsavoury environment of the body piercing studio represents a safer option than the more respectable or cheaper alternatives. The practice of body piercing in the UK remains uncontrolled. PMID- 11485580 TI - Labyrinthine involvement in Behcet's syndrome. AB - We report the neuro-otological findings in 26 consecutive patients with definite and probable Behcet's syndrome unselected for audiovestibular complaints. Auditory and/or vestibular abnormalities were found in 19 (73 per cent) patients, with auditory involvement in 14 (54 per cent) and vestibular in 10 (38.5 per cent) of patients. Peripheral involvement was more common than central involvement for both auditory and vestibular lesions. Bilateral cochlear hearing impairment was the most common audiological finding, whereas unilateral peripheral dysfunction was the prevailing vestibular abnormality. No correlation has been found between audiovestibular lesions and other organ lesions, disease duration or age or sex of the patients. Moreover, there was a lack of interdependence between cochlear and vestibular labyrinthine lesions. We conclude that a full neuro-otological assessment in patients under investigation for Behcet's syndrome may reveal labyrinth involvement in a substantial proportion of patients. In view of the absence of a specific diagnostic test for Behcet's syndrome, audiovestibular lesions may provide further diagnostic support for this disorder. PMID- 11485578 TI - In vivo perimenstrual activation of progelatinase B (proMMP-9) in the human endometrium and its dependence on stromelysin 1 (MMP-3) ex vivo. AB - Most matrix metalloproteinases (MMPs) are secreted as inactive proenzymes. Their expression is well documented in several human tissues, but their activators in vivo are still unknown. To address this question, the activation of progelatinase B (proMMP-9) in the human endometrium was selected as a model system. ProMMP-9 was detected by gelatin zymography in homogenates of fresh endometrial tissue sampled during all phases of the menstrual cycle, whereas its active form was observed only during the late secretory and menstrual phases. Furthermore, proMMP 9 was expressed and activated in endometrial explants sampled outside the perimenstrual phase and cultured in the absence of both progesterone and oestradiol, mimicking the menstrual condition in vivo. Analysis of such tissue cultures by gelatin zymography and Western blotting showed that activation of proMMP-9 depended on a secreted factor and was selectively inhibited by either a synthetic inhibitor of stromelysin 1 (MMP-3) or a monoclonal antibody that specifically blocks MMP-3, thus providing strong evidence for the activation of proMMP-9 in vivo by MMP-3. The activation of proMMP-3 was itself inhibited by a broad-range MMP inhibitor in most cultures, but seemed to involve multiple pathways, implying both serine proteinases and metalloproteinases, which could operate in parallel or sequentially. PMID- 11485581 TI - An isolated and sporadic auditory neuropathy (auditory nerve disease): report of five patients. AB - Five patients of various ages with difficulty in speech discrimination were evaluated. All showed evidence of abnormal auditory brainstem responses (ABRs) beginning with the VIIIth cranial nerve. Broad summating potentials were evoked on their electrocochleograms (EcochGs) and they all exhibited almost normal cochlear outer hair cell function by otoacoustic emissions (OAEs) recordings. Behavioural audiometric testing revealed a mild to moderate elevation of pure tone threshold in all patients. The shape of their pure-tone losses varied, being predominantly low-frequency in four patients (rising slope pattern) and flat across all frequencies in one patient. Speech intelligibility scores of all patients were poor and out of proportion to what would have been expected if threshold elevation of pure-tone was of cochlear origin (i.e. markedly poor scores on the speech audiogram with good scores on the auditory comprehension test). Patients were neurologically normal when the hearing impairment was first manifested. We suggest that this type of hearing impairment is due to an isolated and sporadic disorder of auditory nerve function. It occurs in isolation and does not seem to be part of a generalized neuropathological process. PMID- 11485582 TI - Butylcyanoacrylate tissue adhesive for columellar incision closure. AB - Cosmetic outcome of the columellar incision closure in external rhinoplasty patients has been a subject of discussion. This study was conducted to assess whether tissue adhesives provide an alternative option for sutureless closure of columellar skin incisions for cases utilizing open technique rhinoplastic surgery. One hundred and one patients undergoing external rhinoplasty were randomized to either topical application of butylcyanoacrylate or polypropylene sutures for columellar skin closure. The majority of tension on the wound edges was taken up using 5-0 chromic catgut. Cosmetic outcomes were evaluated by two otolaryngologists independently using visual analogue and Hollander wound evaluation scales in a blinded manner. There was no statistically significant difference in cosmesis between the surgeons' evaluation scores for either type or repair of the columellar incision. Since the tissue adhesive forms its own protective barrier, post-operative care is simplified. Closure with adhesives eliminates the need for post-operative suture removal requiring an extra visit that should lead to more efficient use of physician and patient time. Butylcyanoacrylate performs cosmetically as well as standard suture closure of columellar skin incision used for external rhinoplasty. PMID- 11485584 TI - Endoscopic management of basal encephaloceles. AB - Encephaloceles have been traditionally treated by a transcranial or external approach. The advent of endoscopic sinus surgery has allowed a new intranasal approach for the treatment of basal encephaloceles, minimising patient morbidity. We discuss the classification and presentation of encephaloceles, and present our experience in the repair of six cases. We believe transnasal endoscopic repair is the method of choice in dealing with selected basal encephaloceles. PMID- 11485583 TI - The investigation of nasal septal perforations and ulcers. AB - This is a retrospective review of 74 consecutive patients who presented with a nasal septal perforation or ulcer. The aim of this study was to evaluate the contribution of the investigations used to identify the pathology underlying their septal perforation or ulcer. In the majority of cases there was an anterior (85 per cent) smooth, well-circumscribed septal lesion with an occasional bleeding point. Seventy-seven per cent were less than two cm in diameter. The primary presenting symptom was intermittent epistaxis, with the exception of patients with granulomatous disease, who presented more frequently with nasal obstruction. Thirty-five patients (47 per cent) were idiopathic, 29 (39 per cent) traumatic, eight (11 per cent) inflammatory and two (three per cent) infectious. The nasal septum was biopsied in 71 cases, and histology showed non-specific or non-diagnostic findings in 39 (55 per cent), chronic inflammation in 12 (17 per cent), acute inflammation in six (eight per cent), acute to chronic inflammation in four (six per cent) and histological changes consistent with trauma in nine (13 per cent). The anti-neutrophil cytoplasmic antibody test (ANCA) successfully identified six patients with Wegener's granulomatosis, and the angiotensin converting enzyme test (ACE) supported a clinical diagnosis of sarcoidosis in one patient. The remaining investigation methods (CRP, ESR, chest radiograph, biochemistry and coagulation screen) did not contribute to the diagnosis. In this study, the appearance of a nasal septal perforation or ulcer or its location did not predict the clinical diagnosis. A nasal septal bopsy did not significantly contribute toward the clinical diagnosis. On the basis of these findings, a biopsy of a septal perforation or ulcer is unlikely to contribute to the management of patients unless the supportive tests of ANCA or ACE are abnormal or unless malignancy is suspected. This conclusion is restricted to the management of septal perforations as we have found nasal biopsies of polyps of other lesions to be of value. PMID- 11485585 TI - Mandibular advancement prosthesis: first-line alternative to surgery in snoring. AB - Mandibular advancement prosthesis (MAP) is infrequently used in the UK at present for snoring. First-line measures include dietary and weight modification for those that require it. Where such measures are unlikely to be useful or have already failed, surgery is sometimes utilized as a second-line treatment modality. We evaluate the use of MAP as an adjunct to first-line measures, with emphasis on efficacy, side-effects and patient compliance. Case notes of 30 snorers were reviewed and followed up with a questionnaire. Despite being useful in alleviating snoring, the prosthesis was poorly tolerated. Side-effects include increased salivation, temporomandibular joint pain, intra-oral and myofacial discomfort. Patients who persevered with the prosthesis found the early side effects resolved after a few weeks and snoring reduced. MAP can be used in the initial management of snorers but patients need to be educated and encouraged, especially in the first few weeks. PMID- 11485586 TI - Is reflux noted at diagnostic rigid oesophagoscopy clinically significant? AB - This study assessed the ability of otolaryngologists to diagnose and grade reflux disease at rigid endoscopy. Twenty-one out of 25 senior otolaryngologists who were questioned by means of a telephone survey said that if they find evidence of reflux disease at rigid endoscopy of the oesophagus and larynx, their practice is to place the patient on a proton pump inhibitor for six weeks without requesting pH and manometry studies, and without referral to a gastroenterologist. Over a two year period, 21 patients were diagnosed as having reflux disease at rigid endoscopy. This was based on the finding of fluid and erythema in the aerodigestive tract and upper oesophagus. Subsequent oesophageal pH and manometry was performed. Nine out of 21 patients were confirmed as refluxers. This demonstrated an accuracy of less than 50 per cent when using these findings to diagnose gastro-oesophageal reflux at rigid endoscopy. PMID- 11485587 TI - Epidemiological considerations and clinical features of ENT tuberculosis. AB - Between 1 April 1996 and 30 June 1997, 1003 ear, nose and throat (ENT) outpatients and 340 inpatients diagnosed as having pulmonary tuberculosis were analysed for ENT manifestations of tuberculosis to determine the relationship to sputum positivity, whether any high risk factors exist for the ENT manifestations as compared to other pulmonary tuberculosis patients, and the response to anti tubercular treatment. The commonest ENT manifestation was found to be laryngitis (seven cases), which was more common in pulmonary tuberculosis patients (five out of seven), all except one of whom were sputum negative. All of these patients were defaulters from anti-tuberculosis treatment or relapse cases, and vocal cords were the commonest site of involvement. One case of tuberculous tonsillitis and one case of tuberculous mastoiditis were also noted. The practical implications of an awareness of ENT tuberculosis is a benefit of anti-tubercular therapy and hence conservative management usually suffices. PMID- 11485588 TI - Transtemporal facial nerve schwannoma without facial nerve paralysis. AB - Facial schwannoma is a relatively rare but well documented lesion, presenting either as a mass or with facial nerve symptoms. In this report, an extensive facial schwannoma, extending from the brain stem to the periphery with minimal facial nerve symptoms and normal facial function is presented. PMID- 11485589 TI - Bilateral antrochoanal polyps. AB - An extremely rare case of bilateral antrochoanal polyp in an otherwise fit 12 year-old girl is reported. The antrochoanal polyps were removed by bilateral Caldwell-Luc operation. Histopathological examination showed them to be of benign inflammatory nature. PMID- 11485590 TI - Inflammatory pseudotumour of the nasal cavity and sinuses. AB - The term inflammatory pseudotumour encompasses a very diverse group of conditions, with different sites, whose only common feature is the absence of a specific histological diagnosis. The histopathological and aetiopathogenic data have been derived from experience in the thoracic, orbital and abdominal forms, which are much more frequent than disease affecting the nasal cavities and sinuses. However, there is no evidence that these data are applicable to sino nasal pseudotumour. We present a case of inflammatory pseudotumour of the nasal cavity and a review of the literature (19 cases in 30 years) with respect to the clinical and radiological findings, the natural history and the specific management of disease at these sites. Surgery seems to be the best option for this localization. Corticosteroids and radiotherapy may be of value in cases of residual tumour after the histological confirmation of the diagnosis and the exclusion of neoplasia. PMID- 11485591 TI - Malignant melanoma of the eustachian tube. AB - The case report describes the diagnosis and the management of malignant melanoma of the eustachian tube. This is an exceedingly rare condition that was treated with endoscopic surgery and post-operative radiotherapy. During a 15-month follow up, there was no evidence of recurrence. PMID- 11485592 TI - Leiomyosarcoma of the larynx: emergency tracheostomy. AB - Leiomyosarcoma of the larynx is extremely rare. Histological diagnosis for excluding other tumours remains extremely difficult. A rare case of leiomyosarcoma of the larynx, treated by surgery and radiotherapy, is reported. We discuss the clinical presentation, diagnosis, and treatment of leiomysarcoma with a review of the literature. PMID- 11485593 TI - Malignant triton tumour of the parapharyngeal space. AB - We report the clinical course in a 25-year-old male patient of neurofibromatosis with malignant triton tumour (MTT). Triton tumour is a peripheral nerve sheath tumour with rhabdomyoblastic differentiation. This is relatively rare tumour of head and neck region and only 26 cases have been reported to date. The present case is the first reported MTT of parapharyngeal space, one of the most aggressive course among all head and neck MTTs, resulting in the death of the patient within three months following surgical resection. The aggressive nature of this tumour necessitates adjuvant therapeutic measures in addition to radical surgery in order to obtain better survival rates. PMID- 11485594 TI - A recurrent aphthous stomatitis case due to paediatric Behcet's disease. AB - Behcet's disease is a rare condition in children and characterized by a triad of recurrent aphthous stomatitis, genital ulceration and uveitis. The aetiology is unknown, but systemic vasculitis is the main pathology. The oral lesions resemble Sutton disease, which is another form of recurrent aphthous stomatitis (RAS) that composes approximately 10 per cent of cases and is known as major aphthous stomatitis, can easily be overlooked if the other symptoms are not questioned. We presented a 12-year-old girl with the initial complaint of recurrent aphthous stomatitis. During the follow-up period, we observed the other cardinal signs of the Behcet's disease. PMID- 11485595 TI - Malakoplakia of the oropharynx. AB - We present a case of malakoplakia presenting as a lesion in the oropharynx of an elderly smoker. In this case a clinical diagnosis of carcinoma of the pharynx was made, biopsy revealed the nature of the lesion and management was radically altered. PMID- 11485596 TI - Inflammatory pseudotumour involving the skull base and cervical spine. AB - Inflammatory pseudotumour (IPT) is an idiopathic condition characterized by sclerosing inflammation, which mimics a neoplastic process. IPT involving the skull base and cervical spine is distinctly rare and usually indistinguishable from aggressive neoplasms or infection. We report a case of IPT involving the skull base and cervical spine. Initially the patient complained of headache and hearing loss without other neurological dysfunction. Two cycles of oral systemic steroid therapy resulted in only partial responses. Low dose radiotherapy was followed by quadriparesis as the lesion infiltrated into the cervical vertebral bodies. Subsequently a second course of radiation was administered to the whole cervical spine. Marked improvement was observed clinically and radiologically; however, the patient gradually deteriorated and died of sepsis. Given the aggressive nature of disease and the complications related to the long-term treatment, we suggest that a more aggressive therapeutic approach is suitable in extensive IPT of the skull base. PMID- 11485597 TI - Aspiration pneumonitis following papain enzyme treatment for oesophageal meat impaction. AB - We report a case of aspiration pneumonitis following the use of papain enzyme in a patient with chronic obstructive pulmonary disease with meat impacted in his oesophagus. PMID- 11485598 TI - Isolated hypoglossal nerve palsy due to internal carotid artery dissection. AB - A case of an isolated hypoglossal nerve palsy is reported. The differential diagnosis is discussed, in the context of the requirement for careful scrutiny of the entire course of the hypoglossal nerve on imaging, to detect underlying pathology remote from the tongue, and to avoid unnecessary invasive diagnostic procedures prompted by the appearance of a 'pseudomass' of the weak tongue both clinically and radiologically. PMID- 11485599 TI - Laryngocele and squamous cell carcinoma of the larynx. AB - We present a case report of a lady with a laryngocele and a squamous cell carcinoma of the larynx. The pathogenesis of the relationship between these two entities is discussed and the literature reviewed. This association means a carcinoma must be outruled if a laryngocele is detected clinically or radiologically. PMID- 11485600 TI - Well-differentiated liposarcoma of the epiglottis. AB - Liposarcomas of the larynx are very rare. A review of the English literature revealed only 28 published reports of tumours in this anatomical location. Diagnosis requires a high index of suspicion and careful histologic analysis. We present a case of a well-differentiated liposarcoma of the epiglottis, the tenth reported case at this laryngeal subsite. Initial biopsy specimens showed histological characteristics of a liposarcoma, which facilitated provision of optimal surgical treatment after careful analysis of published literature. PMID- 11485601 TI - Primary non-Hodgkin's lymphoma of the larynx (Stage IE) diagnosed by gene rearrangement. AB - An extremely rare case of primary non-Hodgkin's lymphoma of the larynx (Stage IE) diagnosed by gene rearrangement is reported. The patient was a 76-year-old man with a chief complaint of pharyngeal discomfort. Remission was obtained by excision of the tumour and radiotherapy. Surface phenotypic studies of the laryngeal lesion demonstrated a main population of B-cells expressing L-26, some of the atypical lymphocytes positive with UCHL-1. Genotypic analysis of the specimen disclosed a clonal rearrangement of the immunoglobulin heavy chain with the same rearrangement pattern. These data indicate that this patient had non Hodgkin's lymphoma with diffuse large B-cell type. Gene rearrangement analysis was useful for diagnosis. Diagnostic and therapeutic options are discussed in light of the current literature. PMID- 11485603 TI - A visit to Dr Philippe Contencin, at Hopital Saint-Vincent-de-Paul and Hopital Robert Debre, Paris. PMID- 11485606 TI - Thyroid papillary carcinoma in lateral neck cyst: missed primary tumour or ectopic thyroid carcinoma within a branchial cyst? PMID- 11485607 TI - Family carers and the adult head-injured: a critical review of carers' needs. AB - PRIMARY OBJECTIVE: To appraise recent literature on the topic of perceived needs of family carers of head-injured adult relatives. MEDLINE (1966-2000), EMBASE PSYCHIATRY (1987-1999), CINAHL (1982-January 2000) and PSYCHLIT (1967-1999) databases were searched. MAIN OUTCOMES AND RESULTS: A total of 13 papers were reviewed, of which six employed standardized research methods. The remainder used researcher developed questionnaires. Needs described by carers as important were predominantly related to information. Most unmet needs were linked to the provision of emotional support. There were significant differences in the perceived needs and their fulfilment between parental and spousal carers. In addition, unfulfilled needs could be predicted by behavioural problems in patients. CONCLUSIONS: The design and variation in methodology of most studies reviewed limits the generalization of reported data. Further research is required to fully identify both common and individual needs of family carers of the head injured. PMID- 11485608 TI - Neuropsychological assessment difficulties associated with "hard-to-assess" individuals: a retrospective review. AB - This paper presents the first phase of a wider research programme examining neuropsychological assessment as it applies to individuals with physical and sensory disabilities to such a degree as to make them "hard-to-assess" (HTA). The aim of this paper was to investigate the extent to which physical and sensory disabilities interfered with the neuropsychological assessments of individuals referred following traumatic brain injury. A retrospective review of 288 individuals with brain injury revealed 22 who met the criteria for being HTA on the basis of physical or sensory disabilities. The modifications necessary to standard techniques for assessment of these individuals, and the domains in which assessment was difficult or impossible are discussed. Most HTA were individuals who had impairment in two or three of the visual, motor and speech domains. PMID- 11485609 TI - Psychosocial functioning during the year following severe traumatic brain injury. AB - The psychosocial functioning of a group of 65 adults with severe traumatic brain injury was assessed at 6 months and 1 year post-injury. Aspects of emotional, behavioural, and social functioning were investigated. The prevalence of depression remained constant (24%) over time, although there was some individual variation in the reporting of symptoms. Impatience was the most frequently reported behavioural problem at both assessments. Whilst there was a slight increase in the number of behavioural problems and level of distress reported over time, the most obvious change was in the type of behavioural problems that caused distress. At 1 year post-injury, problems with emotional control were found to be most distressing for the patients. A comparison with pre-morbid social functioning showed the loss of employment to be 70%, 30% returned to live with their parents, and relationship breakdown occurred for 38%. There was also a significant and ongoing decrease in all five aspects of social and leisure activities. PMID- 11485610 TI - Biased responding: a case series demonstrating a relationship between somatic symptoms and impaired recognition memory performance for traumatic brain injured individuals. AB - Biased responding on the Sternberg Recognition Memory Test was observed in four patients with traumatic brain injury. None of these individuals met the Diagnostic and Statistical Manual's (DSM-IV) criteria for malingering. Individual recognition memory scores were high shortly after injury, declined to chance or below at the 6- and 12-month evaluations, and then showed substantial recovery by the 24-month evaluation. Recall memory performance actually declined slightly across this same 2-year period. Recognition memory scores were related to the extent to which the patients endorsed somatic items on the Hamilton Rating Scale for Depression (HAM-D). Poor performance was associated with high somatic scores. The relationship between memory and somatic scores on the HAM-D in this case series suggests that unconscious processes can influence memory performance and, because of this, that clinicians should not use such performance as a primary indicator of malingering. More importantly, biased responding and actual memory deficits may coexist. This is indicated in the current cases by the failure of recall memory to improve during the 2 years these patients were followed. PMID- 11485611 TI - The reliability and validity of the SF-36 health survey questionnaire for use with individuals with traumatic brain injury. AB - In order to examine the reliability and validity of the SF-36 for use with individuals with TBI, the SF-36 and three measures of health-related problems in persons with TBI (BDI-II, TIRR Symptom Checklist, Health Problems List) were administered to 271 individuals without a disability, 98 individuals with mild TBI, and 228 individuals with moderate-severe TBI. Internal consistency (reliability) was demonstrated for all SF-36 scales. Significant correlations were found between the SF-36 scales and the other measures, with stronger correlations emerging in the TBI groups. The TBI groups obtained significantly lower SF-36 scores than the comparison group, and the mild TBI group scored lower than the moderate-severe group. For the most part, the differences between the TBI groups disappeared when BDI-II scores were controlled for. These findings suggest that the SF-36 is a reliable and valid measure for use with persons with TBI. PMID- 11485612 TI - The community integration questionnaire with new scoring guidelines: concurrent validity and need for appropriate norms. AB - OBJECTIVE: To investigate the influence of demographic variables and concurrent validity when new scoring guidelines are used with the Community Integration Questionnaire (CIQ). METHOD: Thirty-three adult outpatients with biopsy confirmed malignant brain tumours were evaluated prior to chemotherapy. In contrast to TBI subjects, the brain tumour sample was older (x age= 46.3 years), better educated (x = 13.7 years); and gender was more evenly represented (16 male, 17 female). RESULTS: Demographic variables mediated CIQ scores. Gender effects were still seen, with women significantly higher on Home Integration (r = 0.40, p < 0.05). Older subjects had lower CIQ scores: CIQ total (r = -0.54, p < 0.005); Home Integration (r = -0.42, p < 0.01); Social Integration (r = -0.44, p < -0.005); and Productivity (r = -0.45, p < 0.005). More education was related to higher CIQ total (r = 0.31, p < 0.05); Social Integration (r = 0.30, p < 0.05); and Productivity (r = 0.35, p < 0.05). Significant relationships were seen between CIQ scores and both the Social Activity and Inactivity subscales of the Chronic Illness Problem Inventory (r = -0.43, p < 0.005 and r = -0.68, p < 0.005, respectively). CONCLUSIONS: CIQ norms for age, education, sex, and marital status are strongly recommended. Moderate concurrent validity remained with new scoring guidelines. PMID- 11485613 TI - Clinical and functional outcome after alcohol neurolysis of the tibial nerve for ankle-foot spasticity. AB - PURPOSE: To report one's experience of using 50-100% alcohol for neurolysis of the tibial nerve in chronic ankle-foot spasticity. METHODS: The records of patients who received alcohol neurolysis of the tibial nerve were retrospectively reviewed. Repetitive monopolar nerve stimulation was used to localize the tibial nerve. Outcome measures included muscle tone as measured by the Modified Ashworth Score (MAS), passive ankle range of motion (PROM), effect on clonus, plantar flexor motor strength, visual gait analysis and use of orthoses. RESULTS: A total of 21 tibial nerves were neurolysed in 18 patients (mean age 38.9 +/- 15.8 years, 12 males, six females). Mean duration post-event was 14.8 +/- 3.9 months. The mean pre-neurolysis MAS was 2.50 +/- 0.77 and this improved to 0.97 +/- 0.88 (p < 0.001) and 0.93 +/- 0.85 (p < 0.001) at 1 and 6 months post-procedure, respectively. Average duration of effect was 10.5 +/- 8.9 months. Eleven out of 12 patients (91.7%) with sustained ankle clonus had complete abolishment lasting 6 months. Mean gain in PROM was 24.6 +/- 16.1 degrees and 32.6 +/- 19.0 degrees at 1 and 3 months post-neurolysis, respectively (p < 0.001, < 0.02). No decrease in motor strength was seen post-neurolysis. All 13 ambulant patients had visible improvements in gait. Complications were transient and included dysesthetic pain (4), sensory loss (1) and distal limb oedema (1). CONCLUSION: Alcohol neurolysis (50-100%) of the tibial nerves is an effective and safe method of managing ankle foot spasticity. PMID- 11485614 TI - Vital capacity following traumatic brain injury. AB - This study was designed to describe lung volume subdivisions in individuals with traumatic brain injury. Records were reviewed for 114 consecutive referrals for motor speech evaluations in a community re-entry residential rehabilitation programme. Obtained measures were vital capacity, inspiratory capacity, and expiratory reserve volume at the time of evaluation. Seventy-nine per cent of subjects evidenced at least moderate deficits in expiratory reserve volume, with 60% evidencing severe deficits. Inspiratory capacity was the least affected, with 61% of subjects evidencing values within normal limits. Possible contributors to reduced expiratory reserve volumes include expiratory muscle weakness due to physical inactivity, inability to voluntarily perform the task, or perceived effort. PMID- 11485615 TI - Polymorphism of CCR5 affecting HIV disease progression in the Japanese population. AB - Among several factors associated with HIV-1 disease progression, genetic polymorphism of CCR2, CCR5, and CXCR4 in HIV-1 infection has been found. Single nucleotide polymorphisms (SNPs) in the CCR2, CCR5, and CXCR4 genes as well as a 32-base pair deletion in the open reading frame of the CCR5 gene are associated with HIV disease progression among Caucasians and African-Americans in North America and Europe. However, in populations other than Caucasians and African Americans, SNPs have not been fully examined. In our study SNPs in CCR2 coding and CCR5 regulatory regions have been examined in 98 Japanese HIV-positive individuals. The alleles of CCR5 regulatory regions at -2135T and -2086G are associated with late onset of AIDS (p < 0.05; odds ratio for the early onset of AIDS, 0.502 and 0.404, respectively). In contrast to this, the allele of CCR5 at 2086A is associated with the early onset of AIDS (p < 0.05; odds ratio for the early onset of AIDS, 2.133). A haplotype including two alleles at -2135G and 2086G is associated with the late onset of AIDS (p < 0.05; odds ratio for the early onset of AIDS, 0.372). Thus we found that a CCR5 SNP and haplotype polymorphism affect HIV disease progression even in the Japanese population. This indicates that the CCR5 genetic polymorphism affecting disease progression should be studied in a wider range of population. PMID- 11485616 TI - Practical prevention of vaginal and rectal transmission of HIV by adapting the oral defense: use of commercial lubricants. AB - HIV is transmitted to 6.4 million human beings per year and the majority of these transmissions are sexual. Condoms are highly effective and are recommended as the primary preventive. However, the fact that there are millions of sexual transmissions each year indicates that many people do not use condoms and that additional preventives are needed. The mechanisms of natural prevention of oral transmission by saliva may be adaptable to the susceptible vagina and rectum. The objective of our study was to reduce the sexual transmission of HIV by mimicking saliva's targeting of the transmitting infected leukocytes and any cell-free HIV in seminal fluid. The previously recommended anti-HIV topical microbicide, nonoxynol-9, has not prevented HIV transmission in humans, probably because it causes mucosal irritation and attracts CD4(+) cells. To identify effective preparations that are nonirritating, we studied the anti-HIV activity of commercially available, over-the-counter (OTC) lubricants and vaginal preparations that are judged safest by the U.S. Food and Drug Administration (FDA), and are nonirritating. The effect of OTC preparations on both the production of HIV by infected leukocytes and cell-free HIV suspended in seminal fluid was measured under simulated in vivo conditions. We surveyed 22 OTC vaginal preparations and excluded those with low inhibitory activity and those that were inhibitory but likely to be irritating. Three included preparations are highly active against both HIV-infected leukocytes suspended in seminal fluid and active against cell-free HIV, under in vitro conditions that simulate in vivo conditions. Since the preparations identified here as anti-HIV substances have the advantages of being widely available, inexpensive, acceptable, in the safest U.S. FDA category, and may be used by recipient women or men, they should be tested in clinical trials to help prevent sexual transmission of HIV. PMID- 11485617 TI - Potent antiretroviral therapy initiates normalization of hypergammaglobulinemia and a decline in HIV type 1-specific antibody responses. AB - Next to a profound T cell immunodeficiency, HIV-1 infection induces activation and dysfunction of B cells, resulting in hypergammaglobulinemia. Whereas T cell immune reconstitution with potent antiretroviral therapy has been extensively documented, limited data are available on B cell immune reconstitution. We studied the effect of potent antiretroviral therapy on antibody titers to the viral proteins gp120 and p24 and on total IgG concentrations. Three retrospectively chosen groups were studied: a successfully treated group, untreated controls, and subjects with virological failure after several months of successful therapy. In the successfully treated group, the median total IgG concentrations normalized, whereas they remained elevated in the untreated group and rebounded after an initial decline in the therapy failure group. The HIV-1 specific antibody titers declined in the successfully treated group and followed the rebound of the HIV RNA levels in the therapy failure group. With potent antiretroviral therapy the hypergammaglobulinemia normalized whereas HIV-1 specific immune responses were weakened. The weakening of antiviral immunity with therapy may be relevant for current attempts to gain immunological control over the virus through structured treatment interruptions or therapeutic vaccinations. PMID- 11485618 TI - Lipid rafts and HIV pathogenesis: host membrane cholesterol is required for infection by HIV type 1. AB - In a previous study we showed that budding of HIV-1 particles occurs at highly specialized membrane microdomains known as lipid rafts. These microdomains are characterized by a distinct lipid composition that includes high concentrations of cholesterol, sphingolipids, and glycolipids. Since cholesterol is known to play a key role in the entry of some other viruses, our observation of HIV budding from lipid rafts led us to investigate the role in HIV-1 entry of cholesterol and lipid rafts in the plasma membrane of susceptible cells. We have used 2-OH-propyl-beta-cyclodextrin (beta-cyclodextrin) to deplete cellular cholesterol and disperse lipid rafts. Our results show that removal of cellular cholesterol rendered primary cells and cell lines highly resistant to HIV-1 mediated syncytium formation and to infection by both CXCR4- and CCR5-specific viruses. beta-Cyclodextrin treatment of cells partially reduced HIV-1 binding, while rendering chemokine receptors highly sensitive to antibody-mediated internalization. There was no effect on CD4 expression. All of the above described effects were readily reversed by incubating cholesterol-depleted cells with low concentrations of cholesterol-loaded beta-cyclodextrin to restore cholesterol levels. Cholesterol depletion made cells resistant to SDF-1-induced binding to ICAM-1 through LFA-1. Since LFA-1 contributes significantly to cell binding by HIV-1, this latter effect may have contributed to the observed reduction in HIV-1 binding to cells after treatment with beta-cyclodextrin. Our results indicate that cholesterol may be critical to the HIV-1 coreceptor function of chemokine receptors and is required for infection of cells by HIV-1. PMID- 11485619 TI - Selective increases in HIV-specific neutralizing antibody and partial reconstitution of cellular immune responses during prolonged, successful drug therapy of HIV infection. AB - Because the immune response to HIV depends on viral gene expression, we examined the HIV-specific immune responses in persons whose viral load after highly active antiretroviral therapy (HAART) was <400 on at least 3 occasions over a 12-month interval. Eleven patients were identified. While there was little change in mean HIV-binding antibody (Ab) titers in this group, two persons mounted increases in HIV envelope-specific binding antibody. Neutralizing antibody (NAb) titers against a panel of HIV-1 primary isolates (BZ167, US1, and CM237) increased post HAART (80% neutralization titer against US1, p = 0.06; against CM237, p = 0.04). The two persons with large increases in binding antibody also had increases in primary isolate NAb. Roughly half of HAART recipients had significant increases in neutralizing antibody to the primary isolates US1 and CM237. Compared with CD4 matched, non-HAART controls, there were significant increases in NAb against the subtype B primary isolate US1 (p < 0.0009); no increases were seen against more easily neutralized primary isolate BZ167. There were no differences after HAART in antibody-directed cellular cytotoxicity (ADCC). HAART resulted in a partial restoration of lymphoproliferative responses to recall antigens (tetanus and diphtheria). New responses developed to HIV Gag p24. No patient responded to HIV Env gp160 or gp120 either before or after HAART. The data underscore the lack of functional reconstitution of HIV-specific, CD4-mediated responses despite durable suppression of viral replication. In the setting of stable anti-HIV Ab levels, the development of increased NAb in certain individuals suggests that control of the virus by HAART may assist in immune control of HIV. PMID- 11485620 TI - Serum concentrations of fibroblast growth factor 2 are increased in HIV type 1 infected patients and inversely related to survival probability. AB - HIV-1-infected patients develop a generalized vasculopathy that is clinically most evident as Kaposi's sarcoma (KS), a multifocally appearing endothelial cell derived tumor. Fibroblast growth factor 2 (FGF-2) is a potent autocrine and paracrine mitogen of endothelial cells and has been implicated in the cell proliferation and angiogenesis observed in KS. Here we determined by ELISA the FGF-2 serum concentrations in different clinical groups of HIV-1-infected patients. AIDS-KS patients (n = 53) and HIV-1-infected patients without KS (n = 39) revealed significantly increased FGF-2 serum concentrations (median, 4.5 and 4.6 pg/ml, respectively), as compared with the healthy control group (n = 22; median, 2.2 pg/ml; p < 0.01). FGF-2 concentrations were highest in untreated HIV 1-infected patients (median, 8.6 pg/ml) and were significantly decreased in patients undergoing antiretroviral therapy (AZT-median, 4.5 pg/ml; HAART-median, 2.5 pg/ml; p < 0.01). In addition, FGF-2 serum concentrations above 5.2 pg/ml were associated with a statistically significant higher risk of death in HIV-1 infected patients. Multivariate analysis showed that this effect is independent of CD4 levels, localization of KS (cutaneous or visceral), AIDS-defining opportunistic diseases, and therapy. Circulating FGF-2 may contribute to AIDS associated vasculopathy and may be a sensitive and easily accessible surrogate marker to determine the survival time of HIV-1-infected patients and the efficacy of antiretroviral therapy. PMID- 11485621 TI - Differential pattern in circulating nitrogen derivatives, lactoferrin, and anti lactoferrin antibodies in HIV type 1 and HIV type 2 infections. AB - HIV-1 infection is associated with a dramatic reduction in antioxidative molecules both at the cellular level and in the circulation. This is particularly so for lactoferrin, an iron-binding protein involved in natural defenses (antimicrobial and antiviral activities, etc.) and found in whole secretions, including milk and mucus. In addition to its ability to chelate iron ions, lactoferrin inhibits hydroxy radical formation and interacts with nitric oxide (NO). Levels of plasma lactoferrin decreased in HIV-1-infected patients in correlation with progression of the disease, and highly specific anti-lactoferrin autoantibodies increased. This profile was specific to HIV-1 infection; it was not found in HIV-2-infected patients. In parallel with the drop in lactoferrin, a marked increase in circulating nitrogen derivatives was observed in HIV-1 infected patients, whereas low levels were found in normal donors and in HIV-2 infected patients. These data suggested hyperstimulation of the NO pathway throughout HIV-1 but not HIV-2 infection. This overproduction of NO could play an important role in the development of AIDS symptoms and signs. PMID- 11485622 TI - HLA-A*26, HLA-B*4002, HLA-B*4006, and HLA-B*4801 alleles predispose to adult T cell leukemia: the limited recognition of HTLV type 1 tax peptide anchor motifs and epitopes to generate anti-HTLV type 1 tax CD8(+) cytotoxic T lymphocytes. AB - Genetic risk for adult T cell leukemia (ATL) has been implicated by ethnic and familial segregation of ATL patients from HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). To clarify the genetic risk for ATL, we characterized HLA class I alleles of ATL patients and analyzed the anchor motifs of HTLV-1 peptides binding to HLA class I molecules, using 291 lines of anti-HTLV 1 CD8(+) cytotoxic T lymphocytes (CTLs) generated in vitro with a total of 165 synthetic peptides for HTLV-1 Tax and Env proteins. Allele frequencies of HLA A*26, B*4002, B*4006, and B*4801 were significantly higher in ATL patients than in HAM/TSP patients and asymptomatic HTLV-1 carriers in southern Japan. CD8(+) CTL analysis revealed the HTLV-1 Tax peptide sequence to completely lack anchor motifs of peptides binding to HLA-A*26,B*4002, and B*4006 molecules but to possess one anchor for HLA-B*4801, while the HTLV-1 Env peptide sequence had many anchor motifs for HLA-A*26, B*4002, B*4006, and B*4801 molecules. Most ATL patients featured heterozygous HLA class I alleles composed of HLA-A*26, B*4002, B*4006, and B*4801, with a lower number of HTLV-1 Tax peptide anchor motifs and epitopes generating anti-HTLV-1 Tax CD8(+) CTLs than individuals possessing other HLA alleles. The relationship between Tax epitope and ATL incidence was verified by the significantly decreased number of HTLV-1 Tax epitopes in ATL patients compared with asymptomatic HTLV-1 carriers (p < 0.01) as well as late onset ATL patients (p < 0.001). These results indicate that HLA-A*26, B*4002, B*4006, and B*4801 alleles predispose to ATL because of the limited recognition of HTLV-1 Tax peptide anchor motifs and epitopes capable of generating anti-HTLV-1 Tax CD8(+) CTLs. PMID- 11485623 TI - Effects of retroviral protease inhibitors on proteasome function and processing of HIV-derived MHC class I-restricted cytotoxic T lymphocyte epitopes. PMID- 11485624 TI - Protection of neutralization epitopes in the V3 loop of oligomeric human immunodeficiency virus type 1 glycoprotein 120 by N-linked oligosaccharides in the V1 region. AB - The V3 region of the human immunodeficiency virus type 1 envelope protein gp120 constitutes a potential neutralization target, but the oligosaccharide of one conserved N-glycosylation site in this region protects it from neutralizing antibodies. Here, we determined whether N-linked glycans of other gp120 domains were also involved in protection of V3 neutralization epitopes. Two molecular clones of HIV-1, one lacking three N-linked glycans of the V1 region (HIV 1(3N/V1)) and another lacking three N-linked glycans of the C2 region (HIV 1(3N/C2)), were created and characterized. gp120 from both mutated viral clones had higher electrophoretic mobilities than gp120 from wild-type virus, confirming loss of N-linked glycans. Wild-type virus and both mutant clones replicated equally well in established T cell lines and all three viruses were able to utilize CXCR4 but not CCR5 as a coreceptor. The induced mutations increased gp120 affinity for CXCR4 but caused no corresponding increase in viral ability to replicate in T cell lines. HIV-1(3N/V1) was neutralized at about 25 times lower concentrations of an antibody to the V3 region than were wild-type virus and HIV 1(3N/C2). Soluble, monomeric gp120 from HIV-1(3N/V1) and wild type virus had identical avidity for the V3 antibody, indicating that the V1 glycans were able to shield V3 only in oligomeric but not monomeric gp120. In conclusion, one or more N-linked glycans of gp120 V1 is engaged in protection of the V3 region from potential neutralizing antibodies, and this effect is dependent on the oligomeric organization of gp120/gp41. PMID- 11485625 TI - Survey of reverse transcriptase from the heterosexual epidemic of human immunodeficiency virus type 1 CRF01_AE in Thailand from 1990 to 2000. AB - Genetic diversity of the HIV-1 envelope gene has shown a steady increase over time in the Thai and other regional epidemics. A serial survey of subtype CRF01_AE polymerase gene (RT) diversity in Thailand was performed, using 48 novel and 15 reported sequences covering the period 1990--2000. These sequences were gathered from individuals whose sole risk factor for infection was heterosexual contact. By contrast to envelope, diversity was low and, despite a 40% increase early in the epidemic, has remained static since 1996. These results indicate that epidemic HIV-1 may be constrained within defined limits of genetic diversity at least in some genomic regions. PMID- 11485626 TI - Infectious DNA clone of HIV type 1 A/G recombinant (CRF02_AG) replicable in peripheral blood mononuclear cells. AB - We constructed an infectious DNA clone of the HIV-1 A/G recombinant 97GH-AG2, which was isolated in Ghana in 1997 and was classified originally as subtype A. By phylogenetic and recombination breakpoint analyses, p97GH-AG2 was grouped in the circulating form of A/G recombinants (CRF02_AG) and was found to contain the least amount of subtype G-derived region among the known CRF02_AG HIV-1 DNAs. This result suggests that CRF02_AG may be a predominant form in Ghana. Virions produced by transfection of p97GH-AG2 into 293T cells grew in phytohemagglutinin stimulated peripheral blood mononuclear cells (PBMCs). 97GH-AG2 also replicated efficiently in CCR5-expressing HeLa cells, MAGIC5, but only weakly in the parent MAGI cells, indicating that 97GH-AG2 uses mostly CCR5 as a coreceptor. Isolation of the first HIV-1 (CRF02_AG) DNA clone that replicates in PBMCs will accelerate the molecular analysis of this subtype. PMID- 11485627 TI - Molecular characterization of HIV type 1 vpu genes from mothers and infants after perinatal transmission. AB - We compared 162 vpu sequences of human immunodeficiency virus type 1 (HIV-1) in peripheral blood mononuclear cell DNA of 6 infected mother-infant pairs after perinatal transmission, and found a 90.12% frequency of intact vpu open reading frames. The heterogeneity of vpu genes between epidemiologically linked mother infant pairs was lower compared with epidemiologically unlinked individuals. However, the variability of vpu genes was higher than that seen for other HIV-1 genes, including vif, vpr, tat, and gag p17 from the same mother-infant pairs. Moreover, the infants' sequences displayed patterns similar to those seen in their mothers. The functional domains essential for Vpu activity, including efficient release of virus particles from infected cells and CD4 degradation, were conserved in most of the sequences. In a phylogenetic analysis, the 162 sequences from 6 mother-infant pairs formed distinct clusters for each mother infant pair sequences and grouped with subtype B sequences. These data support the importance of vpu in HIV-1 replication of mother-infant isolates that are involved in perinatal transmission. PMID- 11485629 TI - Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. I. Methods, safety, and clinical implications. AB - Cystic fibrosis (CF), an autosomal recessive disorder resulting from mutations in the cystic fibrosis trans-membrane conductance regulator (CFTR) gene, is the most common lethal genetic illness in the Caucasian population. Gene transfer to airway epithelium, using adenoviruses containing normal CFTR cDNA, leads to transient production of CFTR mRNA and, in some studies, to correction of the airway epithelial ion transport defect caused by dysfunctional CFTR. Inflammatory responses to the adenoviral vector have been reported, particularly at high viral titers. We evaluated the effects of adenovirus-mediated CFTR gene transfer to airway epithelium in 36 subjects with CF (34 individuals, 2 of whom received two separate doses of vector), 20 by lobar instillation and 16 by aerosol administration. Doses ranged from 8 x 10(6) to 2.5 x 10(10) infective units (IU), in 0.5-log increments. After lobar administration of low doses there were occasional reports of cough, low-grade temperature, and myalgias. At the highest lobar dose (2.5 x 10(9) IU) two of three patients had transient myalgias, fever, and increased sputum production with obvious infiltrates on CT scan. After aerosol administration there were no significant systemic symptoms until the 2.5 x 10(10) IU dose, when both patients experienced myalgias and fever that resolved within 24 hr. There were no infiltrates seen on chest CT scans in any of the patients in the aerosol administration group. There were no consistent changes in pulmonary function tests or any significant rise in serum IgG or neutralizing antibodies in patients from either group. Serum, sputum, and nasal cytokines, measured before and after vector administration, showed no correlation with adenoviral dose. Gene transfer to lung cells was inefficient and expression was transient. Cells infected with the vector included mononuclear inflammatory cells as well as cuboidal and columnar epithelial cells. In summary, we found no consistent immune response, no evidence of viral shedding, and no consistent change in pulmonary function in response to adenovirus-mediated CFTR gene transfer. At higher doses there was a mild, nonspecific inflammatory response, as evidenced by fevers and myalgias. Overall, vector administration was tolerated but transfer of CFTR cDNA was inefficient and transgene expression was transient for the doses and method of administration used here. PMID- 11485630 TI - Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium. AB - A phase I clinical trial was conducted in which recombinant adenovirus containing the cystic fibrosis trans-membrane regulator (CFTR) (Ad2/CFTR) was administered by bronchoscopic instillation or aerosolization to the lungs of cystic fibrosis (CF) patients. In this paper, we evaluate the efficiency of Ad2/CFTR-mediated transduction of bronchial airway cells. The ability of an Ad2/CFTR vector to transduce airway cells was first evaluated in patients to whom the vector was administered by bronchoscopic instillation. Cells at the administration site were collected 2 days after treatment by bronchoscopic brushing. Ad2-specific CFTR DNA was detected in four of five individuals by PCR, and Ad2-specific CFTR RNA was detected in three of five individuals by RT-PCR. Ad2/CFTR-mediated transduction of airway epithelial cells was then determined in CF individuals receiving this vector by aerosol inhalation. Ad2-specific CFTR DNA was detected in 13 of 13 individuals 2 days after aerosolization, and in 3 of 5 individuals 7 days after aerosolization. Ad2-specific RNA was detected in 4 of 13 individuals on day 2, but was not detected in the 5 individuals tested on day 7. The percentage of airway epithelial cells containing nuclear-localized vector DNA was < or =2.4% as determined by fluorescence in situ hybridization (FISH). However, in some cases, a high percentage of nonepithelial mononuclear cells or squamous metaplastic epithelial cells was infected with the adenoviral vector. In conclusion, aerosol administration is a feasible means to distribute adenoviral vectors throughout the conducting airways, but improvements in adenovirus-mediated transduction of airway epithelial cells are necessary before gene therapy for CF will be effective. PMID- 11485631 TI - Regulation of adenovirus-mediated elafin transgene expression by bacterial lipopolysaccharide. AB - Lipopolysaccharide (LPS) is a mediator of inflammatory lung injury. Selective augmentation of host defense molecules such as elafin (an elastase inhibitor with antimicrobial activity) at the onset of pulmonary inflammation is an attractive potential therapeutic strategy. The aim of this study was to determine whether elafin expression could be induced by LPS administered after transfection with adenovirus (Ad) encoding human elafin downstream of the murine cytomegalovirus (CMV) promoter (known to be potentially responsive to LPS). In addition, we aimed to determine the effect of local elafin augmentation on neutrophil migration to the lung. LPS significantly up-regulated elafin expression from pulmonary epithelial cells transfected with Ad-elafin in vitro. In murine airways expression of human elafin was achieved using doses low enough (3 x 10(7) plaque forming units) to circumvent overt vector-induced inflammation. LPS significantly up-regulated human elafin secretion in murine airways treated with Ad-elafin [117 ng/ml in bronchoalveolar lavage fluid (BALF) after LPS administration, 5.9 ng/ml after PBS, p < 0.01)]. Over-expression of elafin significantly augmented LPS mediated neutrophil migration into the airways in vivo (1.30 x 10(6) neutrophils in BALF after Ad-elafin/LPS treatment, 0.54 x 10(6) after Ad-lacZ/LPS (p < 0.05), 0.63 x 10(6) after PBS/LPS (p < 0.05)) and significantly enhanced human neutrophil migration in vitro. These data suggest novel functions for elafin in neutrophil migration, and that judicious selection of promoters may allow single, low-dose adenoviral administration to effect inflammation-specific expression of potentially therapeutic transgenes. PMID- 11485632 TI - Gene transfer of manganese superoxide dismutase reverses vascular dysfunction in the absence but not in the presence of atherosclerotic plaque. AB - Impaired endothelium-dependent vasorelaxation (EDVR) is observed in hypercholesterolemia both in the presence and absence of morphological abnormalities and may be due to superoxide anions. Our aim was to assess the effect of gene transfer of manganese superoxide dismutase (MnSOD) to blood vessels from hypercholesterolemic animals with and without atherosclerotic plaque and to compare the effects of endothelial nitric oxide synthase (eNOS) and MnSOD over-expression on vascular dysfunction in the setting of atherosclerosis. Rabbits received a high-cholesterol diet for 10 weeks, resulting in abnormal EDVR in the absence of plaque in the carotids and the presence of plaque in the aorta. In Group 1, adenoviral vectors encoding MnSOD (AdMnSOD) or beta-galactosidase (Ad(beta)gal) were delivered to the carotid arteries in vivo. Four days later, transgene expression and vascular reactivity were assessed. In Group 2, segments of the aorta were transduced ex vivo with AdMnSOD, AdeNOS or both. Transgene expression and vascular reactivity were assessed 24 hr later. In Group 1, MnSOD expression was detected in AdMnSOD-ransduced vessels and impaired EDVR was reversed in the absence of atherosclerotic plaque. In Group 2 (with atherosclerotic plaque present), MnSOD and eNOS expression were detected by western analysis, and eNOS, but not MnSOD over-expression, improved EDVR whereas simultaneous over-expression of eNOS and MnSOD was no better than eNOS alone. Adenovirus-mediated gene transfer of MnSOD to nonatherosclerotic carotid arteries, but not atherosclerotic aorta, normalizes EDVR. eNOS gene transfer improves EDVR, even in the presence of plaque. PMID- 11485633 TI - Stable integration of transgenes delivered by a retrotransposon-adenovirus hybrid vector. AB - Helper-dependent adenoviruses show great promise as gene delivery vectors. However, because they do not integrate into the host chromosome, transgene expression cannot be maintained indefinitely. To overcome these limitations, we have inserted an L1 retrotransposon/transgene element into a helper-dependent adenovirus to create a novel chimeric gene delivery vector. Efficient adenovirus mediated delivery of the L1 element into cultured human cells results in subsequent retrotransposition and stable integration of the transgene. L1 retrotransposition frequency was found to correlate with increasing multiplicity of infection by the chimeric vector, and further retrotransposition from newly integrated elements was not observed on prolonged culture. Therefore, this vector, which utilizes components of low immunogenic potential, represents a novel two-stage gene delivery system capable of achieving high titers via the initial helper-dependent adenovirus stage and permanent transgene integration via the retrotransposition stage. PMID- 11485635 TI - Impact of laminin 5 beta3 gene versus protein replacement on gene expression patterns in junctional epidermolysis bullosa. AB - Molecular therapy studies to date have examined only a limited number of corrective parameters. To assess more global impacts on cellular gene expression for two major molecular therapeutic approaches, we compared gene versus protein delivery in the human genetic disease junctional epidermolysis bullosa (JEB). Both gene and protein replacement of the laminin 5 beta3 (beta3) adhesion molecule restored normal growth and adhesion to poorly viable JEB cells. Gene expression profiling was then performed using cDNA microarrays. The expression of more genes was normalized after beta3 gene transfer than after protein transfer. As anticipated for beta3 delivery, many of the genes whose expression was restored to the normal range were those encoding adhesion molecules and hemidesmosome components. Although gene transfer normalized the expression of a higher percentage of genes than did protein transfer, neither approach fully normalized expression of all genes examined. In addition, both approaches disrupted the expression of some genes, but protein transfer altered expression of a larger proportion of the genes studied. Our findings suggest that therapeutic gene and protein delivery may exert different effects on gene expression and thus may have implications for the development and analysis of molecular therapies for the treatment of genetic disorders. PMID- 11485634 TI - Promiscuity of pre-mRNA spliceosome-mediated trans splicing: a problem for gene therapy? AB - Trans splicing of messenger RNA has been used in experimental settings to replace mutant RNA sequences. We investigated the feasibility of utilizing trans splicing to replace a mutant RET protooncogene sequence known to inappropriately activate this tyrosine kinase receptor. We constructed a pre-trans-splicing molecule (PTM) consisting of a binding domain complementary to the target intron, the 3' splicing signal sequence (3'ss), derived from adenovirus major late transcript intron 1 and a molecular tag sequence. Accurately targeted trans splicing between the human RET exons and the PTM was demonstrated in NIH 3T3 cells cotransfected with the human RET minigene and the PTM. The efficiency of specific trans splicing was estimated to be no more than 15% in the cotransfection experiment. However, in addition to the targeted trans splicing, nontargeted trans splicing to RET exons was observed. Furthermore, the rapid amplification of 5' cDNA ends (5' RACE) analysis demonstrated that nontargeted trans splicing occurred with endogenously expressed pre-mRNAs in TT cells and that specific trans splicing to RET was a rare event. Attempts to reduce nonspecificity by the addition of a stem loop to the trans-splicing construct designed to suppress nonspecific splicing failed to have the desired effect. These observations suggest that overexpression of a trans-splicing construct containing a 3'ss results in promiscuous trans splicing and raise significant questions about the specificity and usefulness of currently used trans-splicing approaches. In addition, these findings raise the possibility that nonspecific spliced products may be produced by a variety of gene therapy constructs. PMID- 11485636 TI - Human germline gene therapy reconsidered. AB - This paper reevaluates the notion of human germline gene therapy (HGLGT) in light of developments in biomedicine, biotechnology, and ethical and policy analysis. The essay makes the following key points. First, because the distinction among "therapy," "prevention," and "enhancement" is not clear in human genetics, "gene therapy" is an inadequate descriptor of the process and goals of germline genetic alterations. The alternate use of the phrase "human germline genome modification" (HGLGM) could avoid a misleading label. Second, procedures that could be construed as genetic "enhancement" may not be as morally problematic as some have supposed, once one understands that the boundaries between therapy, prevention, and enhancement are not obvious in genetic medicine. Third, HGLGM might be the medically and morally most appropriate way of avoiding the birth of a child with a genetic disease in only a small range of cases. Fourth, there are still many ethical and scientific problems relating to the safety and efficacy of HGLGM. PMID- 11485637 TI - A phase I trial of intralesional rV-Tricom vaccine in the treatment of malignant melanoma. PMID- 11485639 TI - Proceeding of the International Conference on Emerging Infectious Diseases 2000. Atlanta, Georgia, USA. PMID- 11485640 TI - Emerging infectious diseases: a CDC perspective. PMID- 11485641 TI - Emerging viral diseases of Southeast Asia and the Western Pacific. AB - Over the past 6 years, a number of zoonotic and vectorborne viral diseases have emerged in Southeast Asia and the Western Pacific. Vectorborne disease agents discussed in this article include Japanese encephalitis, Barmah Forest, Ross River, and Chikungunya viruses. However, most emerging viruses have been zoonotic, with fruit bats, including flying fox species as the probable wildlife hosts, and these will be discussed as well. The first of these disease agents to emerge was Hendra virus, formerly called equine morbillivirus. This was followed by outbreaks caused by a rabies-related virus, Australian bat lyssavirus, and a virus associated with porcine stillbirths and malformations, Menangle virus. Nipah virus caused an outbreak of fatal pneumonia in pigs and encephalitis in humans in the Malay Peninsula. Most recently, Tioman virus has been isolated from flying foxes, but it has not yet been associated with animal or human disease. Of nonzoonotic viruses, the most important regionally have been enterovirus 71 and HIV. PMID- 11485642 TI - Epidemiology, evolution, and future of the HIV/AIDS pandemic. AB - We used mathematical models to address several questions concerning the epidemiologic and evolutionary future of HIV/AIDS in human populations. Our analysis suggests that 1) when HIV first enters a human population, and for many subsequent years, the epidemic is driven by early transmissions, possibly occurring before donors have seroconverted to HIV-positive status; 2) new HIV infections in a subpopulation (risk group) may decline or level off due to the saturation of the susceptible hosts rather than to evolution of the virus or to the efficacy of intervention, education, and public health measures; 3) evolution in humans for resistance to HIV infection or for the infection to engender a lower death rate will require thousands of years and will be achieved only after vast numbers of persons die of AIDS; 4) evolution is unlikely to increase the virulence of HIV; and 5) if HIV chemotherapy reduces the transmissibility of the virus, treating individual patients can reduce the frequency of HIV infections and AIDS deaths in the general population. PMID- 11485643 TI - Antibacterial household products: cause for concern. AB - The recent entry of products containing antibacterial agents into healthy households has escalated from a few dozen products in the mid-1990s to more than 700 today. Antibacterial products were developed and have been successfully used to prevent transmission of disease-causing microorganisms among patients, particularly in hospitals. They are now being added to products used in healthy households, even though an added health benefit has not been demonstrated. Scientists are concerned that the antibacterial agents will select bacteria resistant to them and cross-resistant to antibiotics. Moreover, if they alter a person's microflora, they may negatively affect the normal maturation of the T helper cell response of the immune system to commensal flora antigens; this change could lead to a greater chance of allergies in children. As with antibiotics, prudent use of these products is urged. Their designated purpose is to protect vulnerable patients. PMID- 11485644 TI - Food safety and irradiation: protecting the public from foodborne infections. PMID- 11485645 TI - Public/private sector partnership for emerging infections. AB - This paper gives examples of public/private partnerships that support research, support drug development and that advance policy development, suggesting that such partnerships can advance our understanding and control of emerging infections. The investment in emerging infectious diseases from government and from industry is currently much larger than that from philanthropy. Nevertheless philanthropy, even with limited dollars, is able to play a catalytic function and provide risk capitol for innovative partnerships and could in the future play an even larger role if the value of such investment is better defined and argued to recruit additional dollars to this area. PMID- 11485646 TI - Adaptation of Bordetella pertussis to vaccination: a cause for its reemergence? AB - In the Netherlands, as in many other western countries, pertussis vaccines have been used extensively for more than 40 years. Therefore, it is conceivable that vaccine-induced immunity has affected the evolution of Bordetella pertussis. Consistent with this notion, pertussis has reemerged in the Netherlands, despite high vaccination coverage. Further, a notable change in the population structure of B. pertussis was observed in the Netherlands subsequent to the introduction of vaccination in the 1950s. Finally, we observed antigenic divergence between clinical isolates and vaccine strains, in particular with respect to the surface associated proteins pertactin and pertussis toxin. Adaptation may have allowed B. pertussis to remain endemic despite widespread vaccination and may have contributed to the reemergence of pertussis in the Netherlands. PMID- 11485648 TI - Toward a national laboratory system for public health. PMID- 11485647 TI - Strengthening capability for malaria research in Africa. PMID- 11485649 TI - Early opportunities for prevention: infections of pregnant women and young infants. PMID- 11485650 TI - Teaming up to prevent foodborne disease. PMID- 11485651 TI - Ethical and legal issues in infectious disease research and control. PMID- 11485652 TI - Injection safety. PMID- 11485653 TI - West Nile virus: a newly emergent epidemic disease. PMID- 11485654 TI - Emerging zoonotic diseases. PMID- 11485655 TI - Electronic laboratory-based reporting: opportunities and challenges for surveillance. PMID- 11485656 TI - Plague in the Americas. PMID- 11485657 TI - Intercontinental transmission of West Nile virus by migrating white storks. PMID- 11485658 TI - Opportunistic infections in persons with HIV or other immunocompromising conditions. PMID- 11485659 TI - International partnerships in infectious diseases research, training, and control. PMID- 11485660 TI - Emerging infectious diseases and the law. PMID- 11485661 TI - Waterborne diseases. PMID- 11485662 TI - Xenotransplantation: benefits and risks. PMID- 11485663 TI - Malaria. PMID- 11485664 TI - Institutional review boards: consideration in developing countries. PMID- 11485665 TI - Antimicrobial resistance. PMID- 11485666 TI - Preventing polio from becoming a reemerging disease. PMID- 11485667 TI - GIDEON: a computer program for diagnosis, simulation, and informatics in the fields of geographic medicine and emerging diseases. PMID- 11485668 TI - Migrating populations--a closer view of who, why, and so what. PMID- 11485669 TI - Emerging infectious disease issues in blood safety. AB - Improvements in donor screening and testing and viral inactivation of plasma derivatives together have resulted in substantial declines in transfusion transmitted infections over the last two decades. Most recently, nucleic acid testing techniques have been developed to screen blood and plasma donations for evidence of very recent viral infections that could be missed by conventional serologic tests. Nonetheless, the blood supply remains vulnerable to new and reemerging infections. In recent years, numerous infectious agents found worldwide have been identified as potential threats to the blood supply. Several newly discovered hepatitis viruses and agents of transmissible spongiform encephalopathies present unique challenges in assessing possible risks they may pose to the safety of blood and plasma products. PMID- 11485670 TI - Emerging infectious diseases among indigenous peoples. PMID- 11485671 TI - Migration, refugees, and health risks. AB - Migration both voluntary and forced is increasing all over the world. People are moving in larger numbers faster and further than at any other time in history. This is happening at a time when many countries are ill-prepared to deal with a changing demography and when policies and attitudes to population movement and immigration are hardening. The health implications of this are many, and, in some cases, illness and death rates associated with migration are exacerbated by a lack of policies needed to make migration a healthy and socially productive process. From a public health point of view, this is having and will continue to have serious ramifications for the people that move, the family they leave behind, and the communities that host the newcomers. PMID- 11485672 TI - Penicillium marneffei infection in patients with AIDS. PMID- 11485673 TI - Adventitious agents and vaccines. PMID- 11485674 TI - ProMED-mail: background and purpose. PMID- 11485675 TI - The Wellcome Trust/Burroughs Wellcome Fund joint program in infectious diseases of the tropical developing world. PMID- 11485676 TI - Yellow fever in Para State, Amazon region of Brazil, 1998-1999: entomologic and epidemiologic findings. AB - Yellow fever (YF) is frequently associated with high severity and death rates in the Amazon region of Brazil. During the rainy seasons of 1998 and 1999, 23 (eight deaths) and 34 (eight deaths) human cases of YF were reported, respectively, in different geographic areas of Para State; most cases were on Marajo Island. Patients were 1 to 46 years of age. Epidemiologic and ecological studies were conducted in Afua and Breves on Marajo Island; captured insects yielded isolates of 4 and 11 YF strains, respectively, from Haemagogus janthinomys pooled mosquitoes. The cases on Marajo Island in 1999 resulted from lack of vaccination near the focus of the disease and intense migration, which brought many nonimmune people to areas where infected vectors were present. We hypothesize that YF virus remains in an area after an outbreak by vertical transmission among Haemagogus mosquitoes. PMID- 11485677 TI - Tuberculosis in North Carolina: trends across two decades, 1980-1999. AB - In North Carolina, we analyzed cumulative data for tuberculosis (TB) from 1980 through 1999 to determine trends in incidence, population subgroups at risk, and implications for health policy- makers. The overall incidence rates declined significantly over the study period (p = 0.0001). This decline correlates strongly with an increase in TB patients receiving directly observed therapy. Males have approximately twice the risk for disease, and persons >65 years of age are at the highest risk. For every Caucasian with TB, six blacks, six Hispanics, and eight Asians have the disease. TB incidence rates are declining in all other population subgroups but increasing in foreign-born and Hispanic persons. PMID- 11485678 TI - Waterborne outbreak of tularemia associated with crayfish fishing. AB - In 1997, an outbreak of human tularemia associated with hare-hunting in central Spain affected 585 patients. We describe the identification of Francisella tularensis biovar palaearctica in a second outbreak of ulceroglandular tularemia associated with crayfish (Procambarus clarkii) fishing in a contaminated freshwater stream distant from the hare-associated outbreak. The second outbreak occurred 1 year after the first. PMID- 11485679 TI - Cholera outbreak in southern Tanzania: risk factors and patterns of transmission. AB - To identify risk factors and describe the pattern of spread of the 1997 cholera epidemic in a rural area (Ifakara) in southern Tanzania, we conducted a prospective hospital-based, matched case- control study, with analysis based on the first 180 cases and 360 matched controls. Bathing in the river, long distance to water source, and eating dried fish were significantly associated with risk for cholera. Toxigenic Vibrio cholerae O1, biotype El Tor, serotype Ogawa, was isolated in samples from Ifakara's main water source and patients' stools. DNA molecular analyses showed identical patterns for all isolates. PMID- 11485681 TI - Measles outbreak in a community with very low vaccine coverage, the Netherlands. AB - A 1999-2000 measles epidemic in the Netherlands started with an outbreak in an orthodox reformed elementary school with 7% vaccine coverage. The overall attack rate was 37%: 213 clinical cases among the 255 participating pupils (response 62%) and 327 household members. The attack rate ranged from 0% for the oldest groups of pupils to 88% for the youngest, who had not been exposed in previous measles epidemics. None of 25 vaccinated pupils had clinical symptoms. Among pupils with clinical symptoms, the self-reported complication rate was 25%. These data confirm that measles infection causes severe disease and that vaccination is the most effective means of preventing the disease and its complications. The data also show that clusters of persons refraining from vaccination interfere with measles elimination even in populations with very high overall vaccine coverage (96%). PMID- 11485680 TI - A one-year study of foodborne illnesses in the municipality of Uppsala, Sweden. AB - Surveillance was enhanced and a retrospective interview study performed in 1998 99 to determine incidence, causes, and costs of foodborne illnesses in Uppsala, Sweden. Sixty-eight percent of the detected foodborne illness incidents were single cases, and 32% were outbreaks. Most (85%) of the incidents came to the attention of the municipal authorities through telephone calls from affected persons. Calicivirus, Campylobacter spp., and Staphyloccocus aureus were the most common etiological agents; meat, meat products, and mixed dishes were the most implicated food categories. The incidence of foodborne illness was estimated to be 38 cases per 1,000 inhabitants per year. The estimated average costs per illness were 2,164 Swedish Krona (SEK) ($246) to society and 500 SEK ($57) to the patient. The annual cost of foodborne illnesses in Sweden was estimated to be 1,082 million SEK ($123 million). PMID- 11485682 TI - Afterthoughts about bovine spongiform encephalopathy and variant Creutzfeldt Jakob disease. PMID- 11485683 TI - Visceral leishmaniasis (kala-azar) outbreak in Somali refugees and Kenyan shepherds, Kenya. PMID- 11485684 TI - Doxycycline and eradication of microfilaremia in patients with loiasis. PMID- 11485685 TI - Bovine spongiform encephalopathy and variant Creutzfeldt-Jakob disease. PMID- 11485686 TI - Mad cow disease. PMID- 11485687 TI - Coronary Artery Disease: Vascular Endothelial Growth Factor and Fibroblast Growth Factor. AB - In the last few years, there have been rapid advances in the application of angiogenic growth factor therapy to clinical settings. In particular, vascular endothelial growth factor and fibroblast growth factor were evaluated in a number of different trials that explored various forms of delivery (systemic vs local, gene vs protein) in various patient populations. Although all Phase I trials reported unqualified successes, the results of Phase II blinded trials were decidedly mixed, calling into question the entire concept of therapeutic angiogenesis. However, with continuing advances in the basic knowledge of new vessel growth in mature tissues and a better understanding of the various issues involved in patient selection, evaluation, and drug delivery, there is hope that growth factor therapy will take its place among other therapeutic modalities available to the clinician. PMID- 11485688 TI - Percutaneous Approach to Achieve Therapeutic Myocardial Angiogenesis. AB - Myocardial therapeutic angiogenesis is a novel approach for treating the large number of patients with advanced symptomatic coronary artery disease not amenable to conventional revascularization approaches. Proof of concept has been demonstrated in animal models of myocardial ischemia, and preliminary results suggest possible benefit in clinical trials of direct surgical (transepicardial) delivery of proteins and recombinant genes encoding angiogenic growth factors. Catheter-based intramyocardial gene delivery strategies have been tested in parallel, with the assumption that the less invasive nature and the potential for multiple treatment sessions would make this technique more applicable. Currently, most of the data relating to the catheter-based transendocardial delivery of angiogensis factors are derived from the experience with a single device (Biosense; Johnson & Johnson, Skillman, NJ). Using this injection platform, catheter-based gene delivery was demonstrated in animals to be safe and feasible, with a transfection yield similar to that achieved by the direct transepicardial approach. At the time of this review, more than 45 patients have been enrolled in Phase I clinical trials that evaluate the safety and feasibility of transendocardial delivery of adenovirus and plasmid-mediated gene transfection as well as cellular elements. PMID- 11485689 TI - Pathology of Direct Myocardial Revascularization. AB - Since the last decade, there has been great interest in creating myocardial channels using energy sources and mechanical devices to enhance tissue perfusion as adjunctive therapy for refractory ischemic coronary artery disease. These studies are predicated on the fact that a portion of myocardium in the failing heart is in a state of hibernation. The creation of these channels may lead to neovascularization capable of reviving hibernating myocytes, thus improving cardiac function. This review discusses the pathology after direct myocardial revascularization in animal models and in humans. The fate of the myocardial channels and the potential mechanisms associated with vascular development are emphasized. PMID- 11485690 TI - Cardiac Magnetic Resonance Imaging for the Assessment of Myocardial Angiogenesis. AB - Research in biology and applications of growth factors in coronary artery disease (CAD) has progressed considerably over recent years. Vascular endothelial growth factor and fibroblast growth factor-2 have been more successful in animal models of myocardial ischemia and Phase I studies than in placebo-controlled trials. However, cardiac magnetic resonance (CMR), with its higher sensitivity and specificity indices for identification of CAD, has not been extensively used in trials of angiogenic therapies. Data in animals and in patients suggest that CMR can reliably identify collateral vessels. Therefore, we hypothesize that CMR may depict collateralization induced by angiogenic therapy better than currently used nuclear perfusion imaging modalities. Versatility of the assessment of myocardial function and perfusion in one imaging session, combined with the noninvasive nature of the test, may considerably lower the cost of clinical trials. Use of CMR-derived surrogate end points may provide better risk stratification and assessment of efficacy in patients receiving growth factor therapy. PMID- 11485691 TI - The Fibroblast Growth Factors and Angiogenesis: Basic Considerations. AB - The fibroblast growth factors (FGFs) are a family of potent heparin-binding polypeptides that influence the growth, differentiation, and migration of many cell types. The FGFs are angiogenic and can induce new vessel growth. Their ability to induce new vessel growth is important in normal embryonic development, physiologic processes, pathologic processes such as malignancy, and many disease states. This article focuses on the basic aspects of FGFs in angiogenesis and discusses early clinical applications in this field. PMID- 11485692 TI - Mechanical Myocardial Injury and Angiogenesis: An Association with Therapeutic Potential for Advanced Ischemic Coronary Artery Disease. AB - The healing response of tissue after mechanical injury is a highly evolved complex response that serves as a natural defense mechanism. Tissue wounds typically heal in a temporal sequence of stages. A vital phase of wound healing is the generation of loose reparative tissue that is neovascularized and rich in angiogenic substrates--that of granulation tissue formation. A therapeutic strategy that has emerged for the treatment of patients with advanced atherosclerotic ischemic coronary disease is to therapeutically manipulate the wound healing process and induce injury in the myocardium to stimulate islands of neovascularization. This paper reviews the response of tissue, particularly the myocardium, to various forms of injury. Also discussed is the emerging hypothesis of a threshold of injury (balancing adequate injury to induce neovascularizatioin versus excessive injury resulting in adjacent myocardial damage with contractile dysfunction without additional angiogenic benefit). Initial animal and human studies from our laboratory and that of collaborators, with a new method of injury-induced angiogenesis referred to as mechanical myocardial channeling, are reviewed. PMID- 11485693 TI - Antiplatelet Therapy in AMI: Combining GP IIb/IIIa Inhibition with Reduced-dose Fibrinolytic Therapy. AB - In the therapy of ST-segment elevation myocardial infarction (MI), the close relationship between early reperfusion of the infarct-related artery and improved outcomes has focused research on improving the speed and efficacy of pharmacologic reperfusion therapy. Recently, it has become appreciated that even among patients who achieve normal epicardial reperfusion after reperfusion therapy, myocardial and microvascular perfusion may be inadequate. Such patients are at increased risk for the development of death and congestive heart failure. The addition of glycoprotein (GP) IIb/IIIa antagonists to standard fibrinolytic therapy has been shown to improve both epicardial and myocardial reperfusion. This article focuses on emerging data regarding the combination of GP IIb/IIa antagonists and standard fibrinolytic agents for the treatment of acute ST segment elevation MI. PMID- 11485694 TI - Improving Endothelial Function with ACE Inhibitors: An Important Therapeutic Target After Coronary Interventions. AB - Adjuvant medical therapy plays a central role in the ongoing care of patients with coronary artery disease after percutaneous revascularization. The cardiac and vascular protective effects of angiotensin converting enzyme (ACE) inhibitors make these agents an attractive choice in this setting, especially in light of the HOPE study results. Mounting evidence suggests that ACE inhibitors exert their favorable effects at the cellular level by antagonizing angiotensin II and enhancing nitric oxide bioavailability and, in the process, restoring normal endothelial function. PMID- 11485695 TI - Moving Up the Coagulation Cascade: Potential Factor X Inhibition for Percutaneous Coronary Intervention. AB - Contrary to recent thinking in platelet biology, which had taken a strong stance on final common pathway concepts, the pharmacologic approach to tempering vascular coagulation has moved upstream to earlier steps in the fundamental process. Because coagulation represents a series of bioamplification sequences, the inhibition of early autocatalytic reactions minimizes the production of downstream substrate. The neutralization of factor X, a pivotal coagulation protease in thrombin generation, represents a particularly attractive target in arterial thrombosis following vessel wall trauma in percutaneous coronary interventions. PMID- 11485696 TI - Transcatheter Closure of Patent Ductus Arteriosus Using the Amplatzer Duct Occluder. AB - The Amplatzer duct occluder (ADO) (AGA Medical Corp., Golden Valley, MN) is designed to meet criteria for an optimal transcatheter technique for closure of a patent ductus arteriosus. It was developed as an alternative to surgical closure and other interventional techniques. It is not technically complex, allows controlled deployment, is retrievable and repositionable, and has a low risk of embolization. The ADO also has very high complete closure rate at 1-month follow up comparable to surgical ligation and division. PMID- 11485697 TI - Summary and Comparison of Patent Ductus Arteriosus Closure Devices. AB - A number of patent ductus arteriosus (PDA) occluding devices have been studied in an attempt to develop a transcatheter method of closure of PDA. Some devices were tested in only animal models, whereas others progressed to clinical trials in human subjects. Some devices have been discontinued, some received approval for general clinical use, and many have not yet received approval by regulatory authorities. No prospective randomized clinical trials have been undertaken and, therefore, data on separate clinical trials are used to determine relative efficacy of the devices. Selection of a method of PDA closure depends largely on its minimal diameter and to some degree on its shape. Silent PDAs do not need occlusion. Very small to small PDAs may be occluded by free or detachable Gianturco coils (Cook Cardiology, Bloomington, IN). Moderate-to-large PDAs require closure by devices, conventional surgery, and videothoracoscopic interruption. The choice in the latter group depends largely on the availability of a given method at a given institution at that particular time. Approval of several of the devices by regulatory authorities may result in conduct of prospective randomized clinical trials and use of a device or method most appropriate to the size and shape of the PDA. PMID- 11485698 TI - How useful is cognitive behavioral therapy (CBT) for the treatment of chronic insomnia? PMID- 11485699 TI - What is the optimal protocol for diagnosis of ectopic pregnancy? PMID- 11485700 TI - Is extended-release oxybutynin (Ditropan XL) or tolterodine (Detrol) more effective in the treatment of an overactive bladder? PMID- 11485701 TI - Can early treatment with atorvastatin (Lipitor) improve the outcome of patients with acute coronary syndromes? PMID- 11485702 TI - Clinical inquiries. At what age do patients no longer need colorectal cancer screening? PMID- 11485703 TI - Clinical inquiries. Should we change formula for a formula-fed infant with persistent spitting up, but with adequate weight gain? PMID- 11485704 TI - The future of family medicine: research. PMID- 11485705 TI - The future of family medicine: clinical practice. PMID- 11485706 TI - Reinventing family practice again. PMID- 11485707 TI - A randomized controlled trial of telephone management of suspected urinary tract infections in women. AB - OBJECTIVE: Although urinary tract infections (UTIs) in otherwise healthy ambulatory women are often managed over the telephone, there has been no systematic evaluation of this approach. Our objective was to compare the outcomes of uncomplicated UTIs in healthy women managed over the telephone with those managed in the office. STUDY DESIGN: We randomly assigned women calling their usual provider with a suspected UTI to receive care over the telephone (n=36) or usual office-based care (n=36). All women had urinalyses and urine cultures. All were treated with 7 days of antibiotics. We compared symptom scores at baseline and at day 3 and day 10 after therapy. We also compared patient satisfaction at the end of the study. The settings were family practices in Michigan. POPULATION: We included healthy nonpregnant women older than 18 years. RESULTS: A total of 201 women with suspected UTIs called their physician. Of these, 99 were ineligible, and 30 declined to participate. The women were young (mean age=36.6 years) and predominantly white (86%). Sixty-four percent of the urine cultures had significant growth of a single organism. We observed no difference in symptom scores or satisfaction. Overall, satisfaction was high. CONCLUSIONS: Short-term outcomes of managing suspected UTIs by telephone appear to be comparable with usual office care. PMID- 11485708 TI - Fluticasone propionate compared with zafirlukast in controlling persistent asthma: a randomized double-blind, placebo-controlled trial. AB - OBJECTIVE: The objective of our study was to compare the efficacy and safety of fluticasone propionate (an inhaled corticosteroid) with zafirlukast (a leukotriene modifier) for persistent asthma. STUDY DESIGN: In this randomized placebo-controlled, parallel-group, double-blind, double-dummy trial, patients underwent an 8- to 14-day run-in period followed by 12 weeks of treatment with inhaled fluticasone propionate (88 mg twice daily by metered-dose inhaler), oral zafirlukast (20 mg twice daily), or placebo. POPULATION: We included a total of 338 persistent asthma patients, 12 years of age or older, using short-acting b2 agonists alone. OUTCOMES: measured Efficacy outcomes included changes in pulmonary function, asthma symptoms, rescue albuterol use, nighttime awakenings due to asthma, and quality of life. Safety outcomes included asthma exacerbations, adverse events, and clinically significant laboratory test results. RESULTS: After 12 weeks of treatment, patients taking fluticasone propionate experienced significantly greater improvements in all clinical parameters (symptom scores, percentages of symptom-free and albuterol-free days, albuterol use, and nighttime awakenings) compared with patients taking zafirlukast (P <.05) or placebo (P <.05). Treatment with fluticasone propionate resulted in significantly greater improvements in pulmonary function compared with zafirlukast (P <.05) or placebo (P <.05). Fewer fluticasone propionate patients (4%) had an exacerbation requiring oral corticosteroids compared with those taking zafirlukast (12%) or placebo (10%). CONCLUSIONS: Inhaled fluticasone propionate is more effective than zafirlukast in controlling asthma symptoms, improving pulmonary function, and improving quality of life for patients who are symptomatic with the use of short-acting b2-agonists alone. PMID- 11485709 TI - Adolescent girls' attitudes toward pregnancy: the importance of asking what the boyfriend wants. AB - OBJECTIVE: We evaluated the factors associated with attitudes toward pregnancy among girls presenting to an adolescent health clinic to better predict which girls are at the highest risk of pregnancy. STUDY DESIGN: We used a cross sectional provider-administered survey design. POPULATION: A total 202 girls aged 13 to 18 years presenting consecutively for reproductive health services to an adolescent care clinic were interviewed about their desire for pregnancy. Girls found to be already pregnant at the initial visit (n=54) were removed from analysis. OUTCOMES: measured The main outcome measured was desire for pregnancy. Subjects were grouped by those desiring pregnancy (n=16), those desiring to avoid pregnancy (n=107), and those ambivalent about pregnancy (n=25). RESULTS: The girls who were ambivalent about pregnancy were not significantly different from the girls desiring pregnancy. In unadjusted analysis, girls desiring pregnancy or who were ambivalent about it were more likely to be Hispanic, unemployed, to not attend school, to live with neither natural parent, and to have lived away from home for more than 2 weeks. In adjusted analysis, the reported attitude of the boyfriend toward having a child was the only significant predictor of adolescent girls&rsquo attitude toward pregnancy. CONCLUSIONS: The best predictor of an adolescent girl&rsquos attitude toward pregnancy is her perception of her boyfriends&rsquo desire for a baby. Primary care providers should include boyfriends in any efforts to delay pregnancy in at-risk adolescent girls. Teenagers who are ambivalent about whether they want to be pregnant do not differ significantly from those desiring pregnancy, and should be considered just as high risk. PMID- 11485710 TI - Predictors of follow-up of atypical and ASCUS papanicolaou tests in a high-risk population. AB - OBJECTIVE: Our goals were to assess adherence to the National Cancer Institute clinical practice guideline for the management of atypical squamous cells of uncertain significance (ASCUS) Papanicolaou (Pap) test results in a community at high risk of cervical cancer. We also hoped to identify predictors of adherence to the guideline. STUDY DESIGN: We used an historical cohort and collected data by chart abstraction. POPULATION: Our study included women receiving care in 7 urban community health centers who had an initial ASCUS or atypical Pap test result in 1996. We excluded women with a history of cervical dysplasia or human immunodeficiency virus infection, yielding a final sample of 387 women. OUTCOMES: measured The outcome measured was the level of adherence to the guideline, defined as falling within 1 of 3 mutually exclusive categories (complete, moderate, or low). RESULTS: Care providers recommended colposcopy after an initial atypical Pap test result in 12% of cases and repeat cytology in 67%. Failure to document a plan for management was found in 19% of cases. Complete adherence was achieved for 27% of subjects, moderate adherence for 28%, and low adherence for 45%. The factors associated with complete versus moderate or low adherence included site of care, description of the abnormality (ASCUS vs atypia), availability of on-site colposcopy, and discussing the plan at a visit. CONCLUSIONS: Adherence with the National Cancer Institute clinical practice guideline in this setting was disappointing and varied substantially by site. Factors amenable to change that may improve follow-up include good communication of results with patients and providing colposcopy at the site of primary care. PMID- 11485711 TI - How physicians diagnose urinary tract infections: the potential influence of laboratory regulations on test availability and use. AB - OBJECTIVE: Physician office laboratory regulations may decrease test availability. We examined the potential effects of regulations on test availability and whether the use of tests in diagnosing uncomplicated urinary tract infections is related to availability. STUDY DESIGN: We performed an analysis of a cross-sectional survey conducted in 1994 and 1995. Test availability and use were determined by physicians' reports. POPULATION: The survey respondents included practicing physicians in 3 specialties (family medicine, general internal medicine, and obstetrics and gynecology) from 4 states: Pennsylvania (which had longstanding office laboratory regulations), and Alabama, Minnesota, and Nebraska (states that were not regulated until the implementation of the Clinical Laboratory Improvement Amendment of 1988). OUTCOMES MEASURED: We determined whether 4 specific tests were available in the office and how the tests were used to diagnose uncomplicated urinary tract infections. RESULTS: Our analysis was based on the responses from the 1898 respondents to the survey. All tests were less commonly available in Pennsylvania; this included the dipstick, microscopic urinalysis, wet prep, and urine culture (odds ratio [OR]=0.20-0.34; all P values < .05). The availability of the microscopic urinalysis and culture increased their use (OR = 4.37 and 2.03, respectively; P=.001). The availability of microscopic urinalysis was associated with a decrease in ordering urine cultures (OR=0.42; P=.001), and the availability of the dipstick was associated with a decrease in the use of both the microscopic urinalysis (OR=0.36; P=.02) and the culture (OR=0.48; P=.05). CONCLUSIONS: We found lower test availability in the state with office laboratory regulations and a decrease in testing when availability is reduced, suggesting that laboratory regulations may influence physicians&rsquo diagnostic approach to urinary tract infections. Further study will be required to determine the level of testing that maximizes patient welfare. PMID- 11485712 TI - Treatment of peptic ulcer disease and nonulcer dyspepsia. PMID- 11485714 TI - Is St. John's wort an effective treatment for major depression? PMID- 11485713 TI - Is ticlopidine more effective than aspirin in preventing adverse cardiovascular events after myocardial infarction (MI)? PMID- 11485715 TI - Is delayed antibiotic prescribing a good strategy for managing acute cough? PMID- 11485716 TI - Do antibiotics improve neonatal outcomes in preterm premature rupture of membranes (PPROM)? PMID- 11485717 TI - Surveillance porn. PMID- 11485718 TI - Work outside the office visit. PMID- 11485720 TI - [Antiganglioside antibodies in peripheral neuropathies]. PMID- 11485722 TI - [Pallidal and subthalamic stimulation in Parkinson's disease: lessons from the unsatisfactory results]. AB - BACKGROUND: Deep brain stimulation is being widely used in advanced Parkinson's disease (PD). Outcome determinants are not known. OBJECTIVE: Some relevant data about outcome may be obtained from the study of patients with unsatisfactory results. PATIENTS AND METHODS: We have retrospectively analyzed the number and causes of unsatisfactory results (insufficient improvement 6 months after the intervention) in 211 patients. RESULTS: Forty patients (18.9%) experienced an unsatisfactory result. Inadequate clinical selection by advanced age, abnormal MRI and levodopa unresponsive symptoms was the main cause in 28 cases. In 11 patients a misplacement of the electrode was reported. In 2 cases no cause was identified. Thirty-four out of the 40 patients corresponded to the early experience of the teams involved in the study. CONCLUSION: We conclude that the main factors responsible for a negative outcome are related to the inadequate clinical selection of the patients and the incorrect placement of the electrode. The experience of the team may also be a key factor. PMID- 11485721 TI - [Antiganglioside antibodies: when, which and for what]. AB - BACKGROUND: We report our experience in the study of antiganglioside antibodies and define their clinical value establishing associations between clinical syndromes and immunological findings. METHODS: We analysed 275 sera: Guillain Barre syndrome (GBS) (78), Miller-Fisher syndrome (MFS) (37), chronic inflammatory demyelinating polyneuroapthy (CIDP) (17), multifocal motor neuropathy (NMM) (42), chronic axonal mixed polyneuropathy (PNP) (54), amyotrophic lateral sclerosis (ALS) (28) and lower motor neuron disease (LMND) (17). We have studied the presence of IgG and IgM antibodies to 9 gangliosides using ELISA and TLC. RESULTS: We have detected anti-GQ1b antibodies in 36/37 (97,3%) of patients with MFS, being undetectable after 4 weeks in 83%. A 34 % (26/78) of patients with GBS were positive for several antiganglioside specificities being GalGalNAc the most frequent (54%). Two out of three sera positive for GD1a corresponded to axonal Guillain-Barre. IgM class anti-GM1 antibodies were positive in 10/12 patients with MMN, while only a 3-9% of patients with ALS, CIDP, PNP and LMND presented antiganglioside antibodies. CONCLUSIONS: Analysis of anti-GQ1b antibodies confirms the diagnosis of MFS, excluding other acute brainstem pathologies and, in this study, detection of anti GD1a antibodies indicates axonal damage in GBS and suggest a worse prognosis. IgM anti-GM1 antibodies are only found in MMN. These findings confirm a disease specific correlation between specific neuropathies and antiganglioside antibodies clinically useful. PMID- 11485723 TI - [Intracranial pressure monitoring and CSF dynamics in patients with neurological disorders: indications and practical considerations]. AB - The study of cerebrospinal fluid (CSF) dynamics is central to the diagnosis of adult chronic hydrocephalus (ACH). At present, many neurology and neurosurgery departments use one or more tests to guide diagnosis of this syndrome and to predict patient response to shunting. In specialised centres, the study of CSF dynamics is combined with continuous intracranial pressure (ICP) monitoring. Determination of several variables of CSF dynamics and definitions of qualitative and quantitative characteristics of ICP can be used to establish whether the hydrocephalus is active, compensated or arrested. CSF dynamics and ICP monitoring can also be used to check the correct functioning of the shunt and can be of use in the clinical management of patients with pseudotumor cerebri. Moreover, ICP monitoring is used to guide the treatment of several acute neurological processes. The aim of this review is to describe the fundamentals of CSF dynamics studies and the bases of continuous ICP monitoring. The advantages and disadvantages of several hydrodynamic tests that can be performed by lumbar puncture, as well as the normal and abnormal characteristics of an ICP recording, are discussed. PMID- 11485724 TI - [Premotor potential (Bereischaftspotential) in writer's cramp]. PMID- 11485725 TI - [Crossed buccofacial apraxia]. AB - Praxis and language are considered right hemisphere functions. Buccofacial apraxia, except few occasions, are seen in right-handed patients with left hemisphere lesions with aphasia. We present a right-handed patient with buccofacial apraxia due to an ischaemic lesion of the right hemisphere seen by MRI. Language was normal and there was not limb apraxia in our patient. It is possible that in some individuals the control of voluntary movements of facial muscles is not associated neither to the control of language neither to the learning of complex movements of limbs. PMID- 11485726 TI - [Primary orthostatic tremor: slow harmonic component as responsible of inestability]. AB - BACKGROUND: Orthostatic tremor (OT) is clinically defined as a tremor of the lower limbs and trunk on walking. It bears a significant functional impairement. Although the term orthostatic tremor was first used by Heilman in 1984, Pazzaglia et al had previously described some cases in 1970. Despite the fact that the pathophysiology of this entity is not fully known, the presence of a central oscillator is generally accepted as being responsible. A high frecuency tremor, between 13 and 18 Hz, constitutes an almost patognomonic finding, and treatment with clonazepam usually improves the symptoms. PATIENT AND METHOD: We present a patient who described his symptoms as "cramps" in lower limbs and trunk on standing up, which were relieved on walking or resting. RESULT: This clinical presentation together with a neurophysiological recording of the tremor showing an activity of lower frequency (8 Hz) combined with the usual higher frequency (16 Hz) and above all the clear amelioration of symptoms when treated with gabapentin, i.e. resolution of the low frequency tremor without changes in the 16 Hz tremor, were the peculiar features of this case which merits discussion. CONCLUSIONS: The slow component of the orthostatic tremor is crucial in this case. The improvement with gabapentin is explained by the disappearance of this slow c PMID- 11485727 TI - [Partial benign crises in adolescence]. AB - The aim of this paper is to present the case of teenage patient with partial seizures fulfilling the criteria of benign partial seizures of adolescence. A 16 year-old male patient had two seizures with a sensory-motor "march" that evolved into a secondarily generalized tonic-clonic seizure on the same day. Several weeks before this event he had had several simple partial sensory seizures. The patient had no previous history of seizures and there was no family history of epilepsy. The neurological examination, EEG and magnetic resonance imaging were normal. The patient was treated with antiepileptic monotherapy during two years. The treatment was gradually tapered and withdrawn over the following six months. He has had no recurrences during the five years of follow-up. The early diagnosis of this entity has the significant prognostic and therapeutic repercussions. PMID- 11485728 TI - [Peripheral neuropathy in the POEMS syndrome]. PMID- 11485730 TI - [Sinusal bradycardia associated with the intracranial expansive process and hypothyroidism]. PMID- 11485732 TI - Gene therapy for human severe combined immunodeficiencies. PMID- 11485733 TI - Autoimmunity and apoptosis: the Crohn's connection. PMID- 11485734 TI - Combination of gene delivery and DNA vaccination to protect from and reverse Th1 autoimmune disease via deviation to the Th2 pathway. AB - Using a combination of local gene delivery and tolerizing DNA vaccination, we demonstrate that codelivery of the interleukin-4 (IL-4) gene and a DNA vaccine encoding the self-peptide proteolipid protein 139-151 (PLP139-151) provides protective immunity against experimental autoimmune encephalomyelitis (EAE). We provide evidence for a mechanism whereby IL-4 expressed from the naked DNA is secreted and acts locally on autoreactive T cells via activation of STAT6 to shift their cytokine profile to T helper 2. We also show that DNA vaccines can be used to reverse established EAE by covaccination with the genes for myelin oligodendrocyte glycoprotein and IL-4. This treatment strategy combines the antigen-specific effects of DNA vaccination and the beneficial effects of local gene delivery. PMID- 11485736 TI - Crystal structure of the IL-10/IL-10R1 complex reveals a shared receptor binding site. AB - Interleukin 10 (IL-10) is a dimeric cytokine that plays a central role in suppressing inflammatory responses. These activities are dependent on the interaction of IL-10 with its high-affinity receptor (IL-10R1). This intermediate complex must subsequently recruit the low-affinity IL-10R2 chain before cell signaling can occur. Here we report the 2.9 A crystal structure of IL-10 bound to a soluble form of IL-10R1 (sIL-10R1). The complex consists of two IL-10s and four sIL-10R1 molecules. Several residues in the IL-10/sIL-10R1 interface are conserved in all IL-10 homologs and their receptors. The data suggests that formation of the active IL-10 signaling complex occurs by a novel molecular recognition paradigm where IL-10R1 and IL-10R2 both recognize the same binding site on IL-10. PMID- 11485735 TI - Enhanced CD4+ T cell proliferation and Th2 cytokine production in DR6-deficient mice. AB - We have found that DR6, a member of the TNF receptor family, is highly expressed in resting T cells and downregulated in activated T cells. DR6-targeted mutant mice were generated and showed normal development. However, DR6(-/-) CD4(+) T cells hyperproliferated in response to TCR-mediated stimulation and protein antigen challenge. Activated DR6(-/-) CD4(+) T cells exhibited upregulated CD25 expression and enhanced proliferation in response to exogenous IL-2 stimulation. In addition, increased CD28 and reduced CTLA-4 expression were observed in these cells. Enhanced Th2 cytokine production by activated DR6(-/-) CD4(+) T cells was associated with the increased transcription factor NF-ATc in nuclei. DR6, therefore, functions as a regulatory receptor for mediating CD4(+) T cell activation and maintaining proper immune responses. PMID- 11485737 TI - Bridging the NFAT and NF-kappaB families: NFAT5 dimerization regulates cytokine gene transcription in response to osmotic stress. AB - The transcription factor NFAT5/TonEBP is evolutionarily the oldest member of the NFAT/Rel family of transcription factors. We show that NFAT5 is uniquely related to NF-kappaB and is the only member of the Rel/NFAT family to be activated by osmotic stress. Like Rel/NF-kappaB proteins but unlike the calcium-regulated NFAT proteins, NFAT5 is constitutively dimeric, and dimerization is essential for DNA binding and transcriptional activity. Using dominant-negative proteins that inhibit NFAT5 dimerization, we show that NFAT5 regulates expression of the TNFalpha and lymphotoxin-beta genes in osmotically stressed T cells. Chromatin immunoprecipitation experiments confirm that NFAT5 binds to the TNFalpha promoter in vivo. We suggest that NFAT5 participates in specific aspects of host defense by upregulating TNF family genes and other target genes in T cells. PMID- 11485738 TI - The impact of duration versus extent of TCR occupancy on T cell activation: a revision of the kinetic proofreading model. AB - The widely accepted kinetic proofreading theory proposes that rapid TCR dissociation from a peptide/MHC ligand allows for stimulation of early but not late T cell activation events, explaining why low-affinity TCR ligands are poor agonists. We identified a low-affinity TCR ligand which stimulated late T cell responses but, contrary to predictions from kinetic proofreading, inefficiently induced early activation events. Furthermore, responses induced by this ligand were kinetically delayed compared to its high-affinity counterpart. Using peptide/MHC tetramers, we showed that activation characteristics could be dissociated from TCR occupancy by the peptide/MHC ligands. Our data argue that T cell responses are triggered by a cumulative signal which is reached at different time points for different TCR ligands. PMID- 11485739 TI - How much TCR does a T cell need? AB - Kinetic features of TCR:MHC/peptide interactions dictate their outcome in vitro, some important parameters of which include the number of molecules engaged and the duration of engagement. We explored the in vivo significance of these findings in transgenic mice expressing TCRs in a quantitatively and temporally controlled manner. As anticipated, reduced TCR levels resulted in attenuated reactivity, but response thresholds were substantially lower than expected-at as low as 1/20th the normal TCR numbers and with no indication of phenotypic skewing at suboptimal levels. We also studied survival of T lymphocytes stripped of their TCRs. Unlike B cells, T cells lacking antigen receptors did not die precipitously; instead, populations decayed gradually, just as previously reported in the absence of MHC molecules. PMID- 11485740 TI - MICA engagement by human Vgamma2Vdelta2 T cells enhances their antigen-dependent effector function. AB - Vgamma2Vdelta2 T cells comprise 2%-5% of human peripheral blood T cells, recognize ubiquitous nonpeptide antigens, and expand up to 50-fold during microbial infection. It is not clear why these Vgamma2Vdelta2 T cells expand only after microbial infection. We show here that the stress-inducible molecule, MICA, is induced on the surface of dendritic and epithelial cells by infection with M. tuberculosis in vitro and in vivo. MICA engagement by the activating receptor, NKG2D, present on Vgamma2Vdelta2 T cells, resulted in a substantial enhancement of the TCR-dependent Vgamma2Vdelta2 T cell response to nonpeptide antigens and protein superantigens alike. Thus, a MICA-NKG2D interaction may be necessary for an effective innate immune response to microbe-associated antigens that also are constitutively present in vivo. PMID- 11485741 TI - MHC class I molecules can direct proteolytic cleavage of antigenic precursors in the endoplasmic reticulum. AB - The large set of peptides presented by MHC (major histocompatibility complex) class I molecules are generated by proteolysis of diverse precursors in the cytoplasm and possibly in the endoplasmic reticulum (ER). To define the potential peptide trimming events in the ER, we analyzed proteolytic products generated in isolated microsomes. The residues flanking the N terminus of the final antigenic peptide were rapidly removed within the microsomes but only in the presence of appropriate MHC molecules. Remarkably, the precursor peptide was bound to the MHC molecules in a distinct conformation and required an aminopeptidase activity to generate the optimal peptide. The MHC molecules are therefore not only the final repositories of antigenic peptides, but they can also direct their excision from longer precursors. PMID- 11485742 TI - Unassembled Ig heavy chains do not cycle from BiP in vivo but require light chains to trigger their release. AB - Unassembled Ig heavy chains are retained in the ER via the binding of BiP to the C(H)1 domain, which remains unoxidized. Interestingly, this domain folds rapidly, albeit nonproductively, when heavy chains are released from BiP in vitro with ATP. The in vivo cycling of BiP from heavy chains was monitored using BiP ATPase mutants as kinetic traps. Our data suggest that BiP does not cycle from the C(H)1 domain of free heavy chains. However, heavy and light chain assembly occurs rapidly and requires the ATP-dependent release of BiP. We propose that BiP's ATPase cycle is stalled or nonproductive when it is bound to free heavy chains. The binding of light chains to the complex reactivates the cycle and releases BiP. PMID- 11485743 TI - The alpha(1,3)fucosyltransferases FucT-IV and FucT-VII exert collaborative control over selectin-dependent leukocyte recruitment and lymphocyte homing. AB - E-, P-, and L-selectin counterreceptor activities, leukocyte trafficking, and lymphocyte homing are controlled prominently but incompletely by alpha(1,3)fucosyltransferase FucT-VII-dependent fucosylation. Molecular determinants for FucT-VII-independent leukocyte trafficking are not defined, and evidence for contributions by or requirements for other FucTs in leukocyte recruitment is contradictory and incomplete. We show here that inflammation dependent leukocyte recruitment retained in FucT-VII deficiency is extinguished in FucT-IV(-/-)/FucT-VII(-/-) mice. Double deficiency yields an extreme leukocytosis characterized by decreased neutrophil turnover and increased neutrophil production. FucT-IV also contributes to HEV-born L-selectin ligands, since lymphocyte homing retained in FucT-VII(-/-) mice is revoked in FucT-IV(-/ )/FucT-VII(-/-) mice. These observations reveal essential FucT-IV-dependent contributions to E-, P-, and L-selectin ligand synthesis and to the control of leukocyte recruitment and lymphocyte homing. PMID- 11485744 TI - MASP-3 and its association with distinct complexes of the mannan-binding lectin complement activation pathway. AB - The mannan-binding lectin (MBL) pathway of complement activation is part of the innate immune defense. The binding of MBL to microbial carbohydrates activates the MBL-associated serine proteases (MASPs), which recruit the complement factors, C4 and C2, to generate the C3 convertase or directly activate C3. We present a phylogenetically highly conserved member of the MBL complex, MASP-3, which is generated through alternative splicing of the MASP-1/3 gene. The designation of MASP-3 as a protease is based on homology to known MASPs. Different MBL oligomers were found to have distinct MASP composition and biological activities. MASP-1, MAp19, and direct C3-cleaving activity are associated with smaller oligomers whereas MASP-3 is found together with MASP-2 on larger oligomers. MASP-3 downregulate the C4 and C2 cleaving activity of MASP-2. PMID- 11485745 TI - Analysis of the relationship between viral infection and autoimmune disease. AB - The clinical association between viral infection and onset or exacerbation of autoimmune disorders remains poorly understood. Here, we examine the relative roles of molecular mimicry and nonspecific inflammatory stimuli in progression from infection to autoimmune disease. Murine herpes virus 1 (HSV-1 KOS) infection triggers T cell-dependent autoimmune reactions to corneal tissue. We generated an HSV-1 KOS point mutant containing a single amino acid exchange within the putative mimicry epitope as well as mice expressing a TCR transgene specific for the self-peptide mimic to allow dissection of two pathogenic mechanisms in disease induction. These experiments indicate that viral mimicry is essential for disease induction after low-level viral infection of animals containing limited numbers of autoreactive T cells, while innate immune mechanisms become sufficient to provoke disease in animals containing relatively high numbers of autoreactive T cells. PMID- 11485746 TI - The Crohn's disease-associated bacterial protein I2 is a novel enteric t cell superantigen. AB - An aberrant T cell response to enteric bacteria is important in inflammatory bowel disease. However, the identity of relevant microbial antigens is unknown. Here, we report the presence of I2, a Crohn's disease-associated microbial gene, in the murine intestine. The I2 protein induced a proliferative and IL-10 response by CD4(+) T cells from unimmunized mice. The I2 response was dependent on MHC class II-mediated recognition but did not require antigen processing. Selective activation was observed for the TCR-Vbeta5 subpopulation. These findings indicate that the I2 protein is a new class of T cell superantigen and suggest that colonization by the I2 microorganism in susceptible hosts may provide a superantigenic stimulus pertinent to Crohn's disease pathogenesis. PMID- 11485747 TI - The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements. AB - The interleukin-2 receptor alpha (IL-2Ralpha) chain is an essential component of high-affinity IL-2 receptors. Accordingly, IL-2Ralpha expression helps to regulate T cell growth and other lymphoid functions. Lineage-restricted and activation-dependent IL-2Ralpha transcription is controlled by three upstream positive regulatory regions (PRRs). We now describe an additional IL-2 response element, PRRIV, within intron 1, in humans and mice. PRRIV activity requires GAS motifs that bind Stat5 proteins and additional upstream HMG-I(Y) binding sites. Moreover, IL-2 induces the binding of HMG-I(Y), Stat5a, and Stat5b in vivo to PRRIV and PRRIII, which also functions as an IL-2 response element. Thus, the IL 2 inducibility of the IL-2Ralpha gene is unexpectedly mediated by two widely separated regulatory Stat5-dependent elements, located both upstream and downstream of the transcription initiation sites. PMID- 11485749 TI - Factors influencing Islet of Langerhans graft function and monitoring. AB - Transplantation of islet of Langerhans represents a viable therapeutic option for insulin-dependent diabetes mellitus. Dramatic progress has been recently reported with the introduction of a glucocorticoid-free immunosuppressive regimen that improved success rate, namely, insulin independence for 1 year or more, from 8% to 100%. The fate of islet grafts is determined by many concurrent phenomena, some of which are common to organ grafts (i.e. rejection), while others are unique to nonvascularized cell transplants, including transplant cell mass and viability, as well as nonspecific inflammation at the site of implant. Moreover, islet grafts lack clinical markers of early rejection, making it difficult to recognize imminent rejection and to implement intervention with graft-saving immunosuppressive regimens. In the present review, we will address the problems influencing islet graft success and the monitoring of islet cell graft function. PMID- 11485750 TI - Functional and morphological graft monitoring after liver transplantation. AB - The development of effective immunosuppressive drugs and the refinement of surgical procedures have led to remarkable improvements in the long-term success of liver transplantation. This procedure is now widely recognised as an effective, preferable therapeutic option for the treatment of end-stage liver disease. The early diagnosis of dysfunction is an indispensable tool for the successful management of the hepatic allograft recipient. Liver function is usually assessed by biochemical and morphological examinations, usually based on coagulation factors (fibrinogen, fibrinogen degradation peptide, factor V, prothrombin time and prolonged thromboplastin time), transaminases, gamma-GT, ALP, bilirubin and lactic acid, and histology. Liver biopsy is usually performed before the implantation of the graft to assess the viability of the liver and following liver transplantation, whenever clinical events warrant it or as part of a routine biopsy schedule. PMID- 11485751 TI - Monitoring "de novo" autoimmune hepatitis (LKM positive) by serum type-IV collagen after liver transplant: a paediatric case. PMID- 11485752 TI - GH/IGF system, cirrhosis and liver transplantation. AB - The GH-related effects are primarily mediated by insulin-like growth factor I (IGF-I), a peptide hormone almost completely produced by the liver. Liver cirrhosis is usually accompanied by a fall in protein turnover. Furthermore, an important consequence of chronic liver disease (CLD) is growth hormone/insulin like growth factor (GH/IGF) axis modification and growth failure. Nutritional status also suffers in this condition, and IGF-I has been proposed as a marker of hepatocellular dysfunction, malnutrition and survival. CLD is characterised by alterations of various clinical biochemistry laboratory parameters. Aminotransferases, bilirubin, plasma proteins, together with prothrombin time and gamma globulins, are usually examined for laboratory diagnostic and/or monitoring purposes. These traditional parameters are also used in the perioperative liver transplantation, but an early signal of graft functioning has still not been established. The aim of the present work is a review of the possibility offered by the clinical biochemistry laboratory GH/IGF investigation in the outcome of liver transplantation. PMID- 11485753 TI - Physiological and clinical implications of proANP(1-98) circulating levels in the perioperative phase of liver transplantation. AB - BACKGROUND: ProANP(1-126), the prohormone synthesized and secreted by atrial myocites, generates an ANP peptide family, the main forms of which are proANP(1 30), proANP(31-67), proANP(1-98) and proANP(99-126). These molecular circulating forms are involved in hemodynamic and electrolyte homeostasis. In cirrhotic patients, volume homeostasis is almost impaired due to abnormal sodium retention, which results in ascites formation and hemodynamic changes, including high cardiac output and low systemic vascular resistance. During liver transplantation, in the anhepatic phase, hemodynamic instability may occur because of decreased venous return due to surgical manipulation of inferior vena cava, considerable blood loss or cross-clamping. Moreover, marked hemodynamic instability is often observed at the reperfusion of the graft. AIMS: The aims of present study are to investigate the changes of ANP during the perioperative phases of Orthotopic Liver Transplantation (OLTx) in end-stage cirrhotic patients. PATIENTS AND METHODS: From July to September 1999, 11 cirrhotic patients undergoing to OLTx were included in the study: seven males and four females (average age 46+/-10.4 years) affected by post-alcoholic cirrhosis [Hypertension 15 (1990) 9], post-hepatitis cirrhosis [D.G. Gardner, M.C. Lapointe, B. Kovacic-Milivojevic, C.F. Deschepper, Molecular analisys and regulation of the atrial natriuretic factor gene, in: A.D. Struphers (Ed.), Frontiers in Farmacology and Therapeutics: Atrial Natriuretic Factor, Blackwell, Oxford, England, 1991, pp. 1-22], Wilson disease [Life Sci. 28 (1981) 89] and polycystic disease [Life Sci. 28 (1981) 89], autoimmune cirrhosis [Life Sci. 28 (1981) 89]. In each patient, a hemodynamic assessment was achieved using a Swan Ganz catheter. Periferical venous samples were performed during and immediately after OLTx for the determination of ANP(1-98) and other biohumoral parameters. RESULTS: Mean ANP(1-98) (pmol/ml mean+/-SD) basal levels resulted higher than that recorded in the group of healthy subjects. A significant correlation between 24-h post-reperfusion ANP and intra-operative RBC and RIS requirement was found (p<0.05). The basal values resulted significantly higher than that observed at phase II degrees (p<0.04) and lower than that at phase VI degrees (p<0.05); the anesthetic induction values were significantly lower than that observed at phase VI degrees (p<0.03). CONCLUSIONS: ANP(1-98) values may represent a useful marker of hemodynamic derangements during and after OLTx. Further clinical correlations will need a larger patient basis. PMID- 11485754 TI - Plasma atrial natriuretic peptide (ANP) fragments proANP (1-30) and proANP (31 67) measurements in chronic heart failure: a useful index for heart transplantation? AB - The family of the atrial natriuretic peptides, proANP fragments and the active alphaANP, is strongly related to heart disease. The aim was to study in CHF subjects the relation of mdANP and NtANP with brain natriuretic peptide (BNP) and with other traditional medical parameters. Sixteen CHF patients (aged 51.9+/-13.7 years) and 16 healthy subjects age matched (50.8+/-5.9 years) were selected. Both NtANP and mdANP were higher in CHF patients than in healthy subjects (1436+/-288 vs. 288+/-22 pmol/l p<0.001 and 2305+/-383 vs. 423+/-65 pmol/l p<0.0001, respectively). BNP in CHF patients was 28.0+/-9 pmol/l (reference values 1.7+/ 1.8 pmol/l). Both NtANP and mdANP demonstrated positive correlation with BNP, p<0.0001 and with left atrial end-systolic volume, p<0.05. BNP correlated with left ventricular mass, p<0.03. In conclusion, plasma NtANP and mdANP analyses are useful laboratory markers in CHF patient investigation and follow up. In particular, they could be employed as non-invasive parameters to follow up worsening of systolic dysfunction until heart transplantation is required. PMID- 11485755 TI - Serum cystatin C in renal transplant patients. AB - Assessment of renal function in clinical medicine is of great importance especially in patients with renal transplants. Cystatin C has the characteristics of an ideal marker to assess renal glomerular filtration rate. Forty patients with renal transplants under steady-state post-transplant conditions were included in the study. Steady-state was defined as lack of acute rejection periods during the last 6 months and stable cyclosporin A medication during the past 4 weeks. Gender was balanced with 20 male and 20 female patients, the mean age was 51+/-14 years, time since transplantation was 5+/-3.5 years. Fifteen percent of the patients suffered from diabetes mellitus. Immunosuppression consisted of cyclosporin A, imuran, and prednisolon. To assess renal function cystatin C, creatinine clearance, serum creatinine, and serum beta2-microglobulin were tested. Creatinine was analysed in serum and urine to calculate the creatinine clearance related to 1.73 m(2) body surface. Cystatin C and beta2 microglobulin were determined by using a particle-enhanced turbidimetric assay. Cystatin C correlated best with creatinine clearance (r=0.66), beta2 microglobulin (0.57), and serum creatinine (0.56). The diagnostic accuracy of cystatin C was significantly better than serum creatinine (p<0.05), but did not differ significantly from creatinine clearance (p=0.73), and beta2-microglobulin (p=0.46). Our data show that patients with renal transplants, cystatin C has a similar diagnostic value as creatinine clearance. However, it is superior to serum determination of creatinine and beta2-microglobulin. Cystatin C allows for rapid and accurate assessment of renal function in patients with renal transplants and is clearly superior to the commonly used serum creatinine. PMID- 11485756 TI - Do serum C-reactive protein measurements help to discriminate episodes of renal dysfunction in patients after renal transplantation? AB - INTRODUCTION: This study investigated whether serial daily measurements of serum C-reactive protein (sCRP) could help differentiate episodes of transplant dysfunction due to rejection, infection, cyclosporine A (CsA) nephrotoxicity, or acute tubular necrosis (ATN) in renal-allograft recipients. MATERIALS AND METHODS: Morning serum was obtained daily from 134 patients during the first 30 days after renal transplantation. All episodes of graft dysfunction were recorded and differentiated with transplant biopsies. CRP concentrations were correlated with post-operative graft function and the various causes of graft dysfunction. RESULTS: All patients showed an increase in sCRP in response to surgery, with a maximum on day 2 after transplantation. The sCRP concentration was significantly higher in patients with delayed graft function (mean 61.50 microg/ml) than in patients with primary graft function (mean 38.01 microg/ml) (p=0.001). Bacterial infections other than asymptomatic bacteriuria (mean sCRP 33.98 microg/ml), interstitial graft rejection (mean sCRP 16.43 microg/ml), and ATN (mean sCRP 30.50 microg/ml) were accompanied by significant increases in sCRP compared with uneventful courses. sCRP was unchanged in the presence of viral infections or CsA toxicity. CONCLUSION: Serial sCRP measurements help to identify renal-transplant dysfunction of different origins. However, rejection, infection and ATN show similar patterns of sCRP increase. Thus, sCRP is unable to discriminate the causes of renal-graft dysfunction. Biopsy remains the gold standard for the differential diagnosis of renal-allograft dysfunction. PMID- 11485757 TI - Assessment of cytomegalovirus infections using neopterin and a new immunoblot. AB - Human cytomegalovirus (HCMV) infections are a major cause of morbidity and mortality in immunocompromised patients despite advances in diagnostic tests and antiviral therapies. The underlying study investigates the diagnostic value of the immune marker neopterin and a recently developed HCMV-specific western blot to detect HCMV infections and to differentiate them into either syndromes or diseases. The mean period of observation was 1428 days. Thirteen HCMV diseases and nine syndromes were diagnosed retrospectively. The first appearance of clinical signs or symptoms was always associated with a marked increase of serum and urine neopterin. The HCMV-specific IgM response followed in the mean 9 days later. Median values and the course of the neopterin levels were significantly higher during the HCMV diseases. In addition, the strength of the humoral immune response was related to the severity of the HCMV infection. Patients with HCMV diseases developed antibodies against a higher number of epitopes. The anti-HCMV IgM response persisted in more than 80% of the patients for longer than 3 years. In conclusion, combining the HCMV-specific western blot and neopterin permit detection of the immune response against HCMV, reflect the severity of the infection and might guide the anti-viral therapy. PMID- 11485758 TI - Differential diagnostic use of interleukin patterns in patients being monitored after transplantation. AB - BACKGROUND AND AIM: We investigated how far the determination of selected interleukins in bodily fluids of patients who had received kidney allografts can help to confirm the diagnosis of complications after transplantations. MATERIALS AND METHODS: Levels of soluble interleukin-2-receptor, interleukin 6 and interleukin 8 were determined in serum and urine of 79 patients. According to the type of diagnosis obtained with histological, serological and microbiological methods and to the clinical course, the groups "stable graft function without complication", "allograft rejection", "cytomegalovirus infection", "systemic extrarenal bacterial infection", "urinary tract infection" and "pyelonephritis" were created. RESULTS AND CONCLUSIONS: The activation of the immune system in different ways depending on the trigger substance (alloantigen, virus, bacterium) and the possibility to differentiate systemic and local processes cause typical patterns of interleukin levels in serum and urine in conjunction with the above mentioned complications after kidney transplantation. Cytomegalovirus infections and systemic extrarenal bacterial infections differ from rejection by the unchanged urine interleukins IL 6 and IL 8, the local urinary tract infections differ from rejection by the unchanged serum interleukins. Acute pyelonephritis differs from rejection by the significantly higher serum IL 6 level. During our daily clinical work, the practical interleukin determinations were proven to be an important tool for early and differential diagnosis of complications after kidney transplantation. PMID- 11485759 TI - Determination of the effects of mycophenolic acid on the nucleotide pool of human peripheral blood mononuclear cells in vitro by high-performance liquid chromatography. AB - Immunosuppressive drugs are needed to prevent the rejection of transplanted organs by the immune system. Immunosuppressive antimetabolites act by interrupting cell metabolism. Their mechanism of action can be studied in vitro by measuring the inhibition of biochemical activities which is reflected by changes in the nucleotide content. In our experiments, human peripheral blood mononuclear cells (PBMC) isolated from healthy volunteers were used. After PBMC stimulation with phytohaemagglutinin (PHA) to mimic activation occurring at a rejection crisis, cells were exposed to varying concentrations of different immunosuppressants (i.e., mycophenolic acid, cyclosporin A and prednisolone) for 68 h at 37 degrees C. Changes in nucleotide content were observed by determining the concentrations of 15 nucleotides using a newly developed HPLC method. The results obtained for mycophenolic acid (MPA; final concentrations in a range between 0.1 and 5 micromol/l), cyclosporin A (CsA; final concentrations between 100 ng/ml and 1 microg/ml) and prednisolone (final concentrations between 0.5 and 10 micromol/l) are given as percentage changes in nucleotide content versus controls and are expressed as mean +/- confidence interval. The possibility of synergistic effects was investigated by incubating the cells with mixtures of all three immunosuppressive drugs varying the amount of mycophenolic acid. In addition, we have shown the effects of MPA/guanosine co-incubation on the intracellular nucleotide levels. Stimulation of peripheral blood mononuclear cells with phytohaemagglutinin led to a significant increase of pyrimidine and purine nucleotides versus control values (100%). Pyrimidine (CTP, UDP, UTP) and purine nucleotides (GDP, GTP, ADP, ATP) were elevated up to 153+/-14% and 142+/ 17%, respectively. Under co-incubation of cells with MPA, the GTP level decreased in a dose-related manner to 56+/-3% of control at a MPA final concentration of 5 micromol/l. Concomitantly, an increase of UTP values to 203+/-18% versus control was observed under co-incubation with 1 micromol/l MPA. Co-incubation of mononuclear cells with guanosine (50 micromol/l) compensated for the effects of MPA on intracellular GTP levels. Combination of MPA, CsA and prednisolone did not alter intracellular nucleotide profiles of PBMC compared to those under MPA incubation alone. The depletion of the guanine nucleotide pool and concomitant increase of uridine nucleotides under the influence of the immunosuppressive drug mycophenolic acid is caused by its inhibitory effects on the key enzyme of de novo purine biosynthesis, inosine 5'-monophosphate dehydrogenase (IMPDH). PMID- 11485760 TI - In vitro effects of mycophenolic acid on the nucleotide pool and on the expression of adhesion molecules of human umbilical vein endothelial cells. AB - The immunosuppressive drug mycophenolate mofetil (MMF) and its active metabolite mycophenolic acid (MPA) selectively inhibit inosine 5'-monophosphate dehydrogenase (IMPDH), and therefore interfere with cellular guanine nucleotide biosynthesis. IMPDH is additionally involved in the synthesis of membrane glycoproteins, some of which are adhesion receptors known to play an active part in the regulation of cell-cell contacts, which are crucial in the process of recruitment and transendothelial infiltration of activated leucocytes in the transplanted organ. As a consequence, MPA leads to a reduction of cellular infiltrates in the course of transplant rejection. In the present study, the effects of MPA on human umbilical vein endothelial cells (HUVEC) are investigated at both molecular and cellular levels. In our experiments, HUVECs are treated with tumor necrosis factor-alpha (TNF-alpha; 10 ng/ml) in order to mimic activation occurring at a rejection crisis. The dose-dependent influence of concomitant incubation with MPA (5-20 micromol/l; 48 h, 37 degrees C, 5% CO2) on their intracellular nucleotide profile is observed by determining the concentrations of purine and pyrimidine nucleotides, using a HPLC method based on solvent generated ion-exchange. The possibility of synergistic effects is investigated by incubating endothelial cells with mixtures of three different immunosuppressants (mycophenolic acid; cyclosporin A, 100 ng/ml; prednisolone, 1 micromol/l)--a combination commonly used after transplantation--varying the amount of MPA (5-20 micromol/l). Stimulation with TNFalpha does not significantly modulate the intracellular levels of nucleotides quantitated. In the presence of MPA concentrations of at least 5 micromol/l, GTP levels (68+/-12%) are significantly decreased compared to controls (100%). At a concentration of 20 micromol/l MPA, the GTP amount is reduced to 58+/-7%. In contrast to these observations, the levels of UDP and UTP are increasing significantly under coincubation with MPA concentrations greater than 5 micromol/l. At 20 micromol/l MPA, UDP and UTP are increased to 147+/-19% and 114+/-11%, respectively. All other nucleotides (CTP, ADP, ATP) reveal no significant alterations in their intracellular concentrations under the conditions applied. Incubation of TNFalpha treated HUVEC monolayers, with a mixture of three immunosuppressive drugs varying the amount of MPA, show no significant differences compared with the data observed after incubation with MPA alone. In addition, the influence of MPA (10 micromol/l) on a cellular level is observed by measuring the cell surface expression of adhesion molecules on cytokine-stimulated HUVECs, using TNFalpha (10 ng/ml), interferon-gamma (100 ng/ml), interleukin-1beta (10 ng/ml) and interleukin-8 (20 ng/ml). Expression of the intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), endothelial leucocyte adhesion molecule-1 (ELAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1) was assessed by flow cytometry. Activation of endothelial cell monolayers with TNFalpha significantly increases the mean fluorescence intensity of VCAM-1 (361+/-14%) and ICAM-1 (429+/-47%) surface expression, compared to controls, and additionally induces E-selectin expression (2919+/-134%). The same tendencies, but in a lesser degree, are observed under stimulation of cells with either IFNgamma or IL-1beta. Incubation with a combination of TNFalpha and MPA leads to a significant reduction in VCAM-1 (329+/-13%) and E-selectin (2613+/ 167%) expression, compared to the values obtained for HUVEC incubated with the cytokine alone. Treatment of the cells with IL-1beta/MPA also reduces the expression of VCAM-1 to a level significantly lower than the level observed after stimulation with IL-1beta. Incubation with MPA alone reveals no significant modulation in the expression of all surface molecules tested compared to the values of unstimulated HUVECs. The experiments show that the immunosuppressive action of MPA not only inhibits lymphocyte proliferation but also decreases the expression of adhesion molecules on endothelial cells, which are the first target of the cellular rejection process. PMID- 11485761 TI - Sialic acid in normal human tear fluid. AB - PURPOSE: We measured the concentration of sialic acid, the terminal component of mucin, in normal diluted human tears. METHODS: Twenty-microliter tear samples were collected from 31 healthy volunteers (average age = 50.7 years) using micropipette after 50 microL instillation of saline. We investigated the correlation of concentration between glycoprotein and sialic acid and the difference between the right and the left eyes, as well as the reproducibility of the sampling procedure. RESULTS: There was significant correlation of the concentration between glycoprotein and sialic acid (right eye: r = 0.952, P <.0001; left eye: r = 0.976, P <.001). There was no significant difference in concentration between the right and the left eyes. Also, the reproducibility was considered acceptable in three measurements of sialic acid using the present procedure. The sialic acid concentration in normal diluted tears was 37.1 microg/mL on average. CONCLUSION: Our data indicates that sialic acid concentration is an indicator for the concentration of glycoprotein and that this method of measurement is applicable to the analysis of mucin-deficient disorders. PMID- 11485762 TI - Immunosuppressive effect of cholera toxin B on allergic conjunctivitis model in guinea pig. AB - PURPOSE: To investigate the new method of immunotherapy using cholera toxin B (CTB) in experimental allergic conjunctivitis. METHODS: We used 21 white Hartley guinea pigs. The animals were sensitized by intraperitoneal injection of ovalbumin (100 microg/mL) and albumin hydroxide (5 mg/mL) repeated after an interval of 2 weeks. One week after the second injection, conjunctivitis was induced by topical instillation of ovalbumin (5 mg/mL). The animals were divided into two groups, CTB group and control group. The CTB group underwent pretreatment of topical instillation of CTB (4 microg/30 mL) and ovalbumin (10 microg/30 mL), three times a day for 3 days, 1 week before the intraperitoneal injection. The control group did not undergo the pretreatment. Clinical examination was performed at 0.5, 6, and 24 hours after the development of conjunctivitis. Histological examination was performed at 6 and 24 hours. RESULTS: Both groups developed palpebral and bulbar edema with hyperemia 30 minutes after instillation of ovalbumin. The allergic reaction score was significantly less in the CTB group than in the control group (Mann-Whitney U test: P <.01). The clinical reactions subsided after 6 hours. The CTB group showed less eosinophilic infiltration in the conjunctiva and the limbus, particularly in the conjunctival epithelium, than the control group at 6 and 24 hours. CONCLUSION: Pretreatment with topical CTB and antigen suppresses clinical and histological findings in experimentally induced allergic conjunctivitis. PMID- 11485763 TI - The optimal molecular weight of dispersive type sodium hyaluronate for the reduction of corneal endothelial damage induced by sonication, irrigation, and aspiration. AB - PURPOSE: The purpose of this study was to investigate the optimal molecular weight of dispersive sodium hyaluronate (Na-HA) for the reduction of corneal endothelial damage induced by sonication, irrigation, and aspiration, using enucleated pig eyes. METHOD: The phaco-needle of phacoemulsification and aspiration (PEA) equipment was inserted into the anterior chamber after aqueous humor replacement with a 1% Na-HA solution of various molecular weights (2420 x 10(3), 1460 x 10(3), 1100 x 10(3), 520 x 10(3) or 290 x 10(3)). Then sonication, irrigation, and aspiration were conducted for 60 seconds. The residual rate of Na HA in the anterior chamber and the damaged area of corneal endothelium were determined using an image analyzer. RESULTS: Na-HA with a molecular weight of 1100 x 10(3) gradually disappeared from the anterior chamber after mixing with the irrigating solution, and the damaged area in only the 1100 x 10(3) group was significantly smaller compared with that in the control group. CONCLUSION: These results suggest that an optimal molecular weight exists for dispersive hyaluronate applied for the protection of intraocular tissues during PEA. Under the conditions of this study, Na-HA with a molecular weight of 1100 x 10(3) displayed the highest protective efficacy. PMID- 11485764 TI - The effect of polymethylmethacrylate and acrysof intraocular lenses on the posterior capsule in patients with a large capsulorrhexis. AB - PURPOSE: We have previously shown that patients who have a capsulorrhexis larger than the diameter of a polymethylmethacrylate (PMMA) intraocular lens (IOL) rapidly develop increased posterior capsule opacification (PCO), in effect, producing an example of enhanced PCO. This study focuses on the influence of AcrySof IOLs on this process. METHODS: Phacoemulsification was performed on two groups of patients. The first consisted of 38 patients with a large capsulorrhexis of 6-7 mm who received a 5.5-mm PMMA IOL. The second group of 32 patients had identical surgery and a 5.5-mm MA30 AcrySof IOL was implanted. On days 1,14, 28, 90, 180, and 360, high resolution digitized retroillumination images were taken of the posterior capsule. The PCO area was measured by image analysis at 90, 180, and 360 days. Wrinkling of the posterior capsule was determined at 90 days, and the progression or regression of lens epithelial cell (LEC) proliferation was established by examination of serial images at 28 and 180 days. RESULTS: At 90 days, 79% of the patients with PMMA IOLs had moderate to severe wrinkling of the posterior capsule, whereas the patients with AcrySof IOLs had none (P <.001). The percentage of PCO area was 69% for the PMMA IOLs and 24% for the AcrySof IOL group at 360 days (P <.0001). In the PMMA group, LEC progression occurred in 77%, LEC growth was stable in 15%, and LEC regression occurred in only 8%, compared to 69% of patients with AcrySof IOLs (P <.0001). CONCLUSIONS: In patients with a rhexis larger than the IOL, AcrySof IOLs potentially can prevent capsular wrinkling and cause less PCO than a PMMA IOL with a similar rhexis size. The LEC regression occurs with AcrySof between 28 and 180 days. The reasons for this are discussed. PMID- 11485765 TI - Peripherin/RDS gene mutation (Pro210Leu) and polymorphisms in Japanese patients with retinal dystrophies. AB - PURPOSE: To determine the frequency of peripherin/RDS (retinal degeneration slow) gene mutations in Japanese patients with retinal dystrophies. METHODS: We analyzed the peripherin/RDS gene in 54 unrelated Japanese patients with retinal dystrophies. Genomic DNA was amplified by polymerase chain reaction (PCR) and the PCR products were sequenced. We also examined 100 healthy subjects, seeking mutations or variations of the peripherin/RDS gene. RESULTS: Of the 54 Japanese patients, one with retinitis pigmentosa had a heterozygous C to T change at the second nucleotide at codon 210 of exon 2 (CCT to CTT/Pro210Leu) of the peripherin/RDS gene. None of the 100 individuals with normal fundi had the Pro210Leu mutation of the peripherin/RDS gene. Three variants of the peripherin/RDS gene (GTC to GTT/Val106Val, Glu304Gln, and Gly338Asp) were also found. The first variation (GTC to GTT/Val106Val) was silent. Two concurrent missense variations (Glu304Gln and Gly338Asp) were seen in 25.9% of the affected patients and in 29% of the healthy individuals. CONCLUSION: A novel mutation (Pro210Leu) of the peripherin/RDS gene has been found in one Japanese patient with retinitis pigmentosa. The alterations of Val106Val, Glu304Gln, and Gly338Asp may be polymorphic variants in the Japanese population. PMID- 11485766 TI - Antithrombotic effect of ticlopidine in an experimental model of retinal vein occlusion. AB - PURPOSE: Ticlopidine inhibits adenosine diphosphate (ADP)-induced platelet aggregation and may be effective in patients with retinal vein occlusions (RVO). This study tests the efficacy of ticlopidine in an animal model of RVO. METHODS: Rose bengal-mediated argon laser photothrombosis of retinal veins was created in rabbits pretreated with oral ticlopidine, aspirin, or saline. The number of laser spots necessary to produce a partial or complete RVO was recorded and tabulated. RESULTS: Pretreatment with ticlopidine significantly increased the number of laser spots needed to produce a partial (P =.02), or a complete (P =.002) RVO as compared to the control group. Pretreatment with ticlopidine significantly increased the number of laser spots needed to produce a partial RVO (P =.02). Pretreatment with aspirin significantly increased the number of laser spots needed to produce a complete RVO (P =.002). CONCLUSION: Ticlopidine may be a useful antiplatelet agent for the treatment of patients with RVO. Patients treated with ticlopidine should be monitored for the possible development of hematologic disorders. PMID- 11485767 TI - Cell adhesion glycoproteins in the human lamina cribrosa. AB - PURPOSE: The distribution of the cell adhesion glycoproteins, laminin, fibronectin, tenascin, vitronectin, thrombospondin, and entactin/nidogen, was examined in the human lamina cribrosa. METHODS: Frozen sections of the optic nerve head from 7 normal human elderly donors were stained by immunohistochemistry. RESULTS: All six glycoproteins were detected in this tissue. While laminin and entactin/nidogen were observed linearly, reflecting the localization of basement membranes, fibronectin was identified diffusely. Marked tenascin immunoreactivity was apparent in the lamina cribrosa, but little or no tenascin staining was detected in the sclera. Vitronectin showed a fine fibrillar staining pattern in the lamina cribrosa, and, to a lesser extent, in the sclera and pial septa. Thrombospondin staining was apparent only in the sclera and the lamina cribrosa, which traversed the optic nerve. CONCLUSIONS: These results indicate that extracellular matrix components in the lamina cribrosa differ from those in the sclera or pial septa. This study is the first report that the human lamina cribrosa includes vitronectin and thrombospondin. PMID- 11485768 TI - Indocyanine green angiograms of choroidal nevi. comparison between confocal and nonconfocal scanning laser ophthalmoscope and fundus video camera. AB - PURPOSE: A fundus video camera and a nonconfocal scanning laser ophthalmoscope (SLO) detect direct light and indirect light, whereas a confocal SLO detects mostly direct light. Differences in confocal and nonconfocal SLO images and fundus video camera images are most likely due to their different optical systems. These differences were examined in indocyanine green (ICG) angiograms of a choroidal nevus. METHODS: A confocal SLO, a nonconfocal SLO, and a high resolution digital fundus video camera were used to obtain ICG angiograms of pigmented choroidal nevi in 4 patients for 30 minutes following dye injection. RESULTS: All the angiograms showed a hypofluorescent region in the nevus until 10 14 minutes after dye injection, except in 1 patient in whom no hypofluorescent region was seen in an early confocal-SLO angiogram. From 20 minutes to 30 minutes postinjection, the hypofluorescent regions were still visible in all fundus video camera angiograms and nonconfocal SLO angiograms but not in confocal SLO angiograms. CONCLUSIONS: Early angiograms taken with the three angiography systems showed a similar appearance of the choroidal nevus. However, late ICG angiograms with a confocal SLO showed different images from those taken with a nonconfocal SLO or a fundus video camera. It is suggested that the angiography system and the aperture size of an SLO should be selected according to the aspect of the pigmented choroidal nevus that is of interest in late-phase ICG angiography. PMID- 11485769 TI - Ligneous conjunctivitis: a case report. AB - BACKGROUND: Ligneous conjunctivitis is a rare condition characterized by chronic, recurrent conjunctivitis associated with pseudomembrane, and it may involve other mucous membranes in the mouth, nasopharynx, trachea, and vagina. We examined and treated a case of presumed ligneous conjunctivitis. CASE: The patient was a 10 year-old boy. His chief complaints were visual impairment, discomfort, and discharge, but no itching in his eyes. His upper eyelids appeared thick without swelling. He had a past history of surgery for lid entropion. His two siblings had similar follicular conjunctivitis. OBSERVATIONS: This case exhibited several characteristics of ligneous conjunctivitis, such as large follicles, recurrent pseudomembrane and normal level IgE in the serum. Indispensable characteristics of vernal keratoconjunctivitis, strong itching, and extensive papillary formation, were not found. In spite of the lack of woody hardness of the conjunctiva, other clinical findings led to the diagnosis of ligneous conjunctivitis. Definite histological diagnosis was not obtained, because of the lack of common histological characteristics among previously reported cases with ligneous conjunctivitis. The boy had developed corticosteroid glaucoma after instillation of dexamethasone 0.1% for 7 months at a previous time. We successfully treated this case with combined instillation of fluorometholon and cyclosporin after trabeculotomy. CONCLUSIONS: Ligneous conjunctivitis must be considered as one type of differential diagnosis of vernal keratoconjunctivitis. Cyclosporin is an effective alternative for the treatment of ligneous conjunctivitis, especially in a case with a possible history of corticosteroid glaucoma. PMID- 11485770 TI - Long-term outcome of systemic cyclosporine treatment following penetrating keratoplasty. AB - PURPOSE: To perform a retrospective study to evaluate the long-term outcome of systemic cyclosporine treatment as an adjunct to topical corticosteroid treatment after penetrating keratoplasty (PKP). METHODS: Twenty-six high-risk patients (27 eyes) who received systemic cyclosporine following PKP for an average of 5.4 months were compared with another series of 57 patients (57 eyes) who did not receive cyclosporine after PKP. RESULTS: Endothelial rejection developed in 2 cases during cyclosporine treatment and in 6 cases after discontinuation. The rate of rejection-free graft survival was similar between the treated and the control groups. The control group showed a significantly higher rate of graft survival than the treated group. As side effects in the treatment group, transient elevation in blood urea nitrogen or creatine developed in 7 cases. Increase in glutamate oxaloacetate transaminase (GOT) or glutamate pyruvate transaminase (GPT) developed in 4 cases. Severe side effects were absent throughout the series in both groups of patients. CONCLUSION: Systemic cyclosporine treatment for several months did not reduce the incidence of rejection nor improve the rate of graft clarity in the long term in high-risk patients after PKP. PMID- 11485771 TI - Current trends in cataract and refractive surgery in Japan: 1999 survey. AB - PURPOSE: The eighth annual survey was carried out by mail in February 2000 to investigate the current trends in cataract and refractive surgery in Japan. RESPONDENTS: Questionnaires were sent to 930 ophthalmologist members of the Japanese Society of Cataract and Refractive Surgery. Data received from 457 (49.1%) of the recipients were cross-analyzed and compared with those from the previous surveys. RESULTS: In cataract surgery, 17% of respondents were doing 51 or more cases per month, 94% preferred phacoemulsification, 58% employed the self sealing wound closure technique, and 26% used topical anesthesia for phacoemulsification. In refractive surgery, excimer laser surgery and astigmatic keratotomy attracted notably high interest, while less attention was directed toward radial keratotomy and intrastromal corneal ring. Laser in situ keratomileusis, photorefractive keratectomy, phakic intraocular lens, intrastromal corneal ring, and radial keratotomy were judged to be useful refractive surgical procedures by 69.0%, 40.0%, 24.6%, 14.2%, and 8.0% of the respondents, respectively. CONCLUSION: There are trends toward more surgical procedures performed by a surgeon, shorter period of hospitalization, and increasing preference for small incision cataract surgery. Refractive surgery is not yet widely performed, but laser in situ keratomileusis is viewed as the most promising procedure. PMID- 11485772 TI - Systemic circulatory parameters. comparison between patients with normal tension glaucoma and normal subjects using ambulatory monitoring. AB - PURPOSE: To compare circadian changes of systemic circulation in patients with normal tension glaucoma (NTG) and normal subjects. METHODS: Forty-three patients with NTG and 226 normal subjects were enrolled in this study. Circulatory parameters, including blood pressure (BP) and pulse rate (PR), were measured in all subjects for 49 hours using an ambulatory monitoring system. In addition to a comparison between NTG patients and normal controls, the same parameters were compared between NTG patients who had progressive field defects and NTG patients who had stable field defects. RESULTS: The BP in NTG patients was significantly higher than in normal subjects. The nocturnal dip of BP in NTG patients was similar to the dip in normal subjects. However, the BP dip in NTG patients showing progressive visual field defects was significantly smaller than in patients with NTG showing stable visual field defects. Blood pressure fluctuation in sleep in the "progressive" patients was significantly greater than in the "stable" patients. Patients with NTG whose random BP was in a normal range showed a higher BP than normal subjects. The dip in PR in NTG patients was significantly lower than in normal subjects. CONCLUSIONS: An insufficient physiological nocturnal BP dip or a greater nocturnal fluctuation in BP may disturb the microcirculation of, and/or may directly damage, the optic nerve, resulting in increasing field loss in NTG. PMID- 11485773 TI - Studies on vitrectomy cases associated with complicated branch retinal vein occlusion. AB - PURPOSE: To study the preoperative factors leading to vitrectomy, and to demonstrate the postoperative conditions causing the decreased visual acuity that results from branch retinal vein occlusion (BRVO). METHODS: In 113 patients (114 eyes) with BRVO, the following data were analyzed: age, general complications, distribution of occluding vessels, location of retinal breaks, classification of vitreoretinal pathology, and the number of cases, period from onset of BRVO to vitreous hemorrhage and from vitreous hemorrhage to vitrectomy, number of operations, relationship between posterior vitreous detachment (PVD) and number of operations, preoperative photocoagulations, pre- and postoperative visual acuity, and cases with poor visual outcome. RESULTS: The visual prognosis was much better in cases with vitreous hemorrhage only than in those with proliferative membrane and retinal detachment (P =.0023). Repeated surgeries were needed in the cases where there was only partial PVD (P =.0029). Macular disorders and optic nerve atrophy were the main causes of postoperative visual acuity < 0.1. CONCLUSIONS: Early vitrectomy before development of vitreo retinal proliferation and retinal detachment, especially in cases where there is only partial PVD, seems to be essential for case management and treatment to attain better visual acuity for the patient. PMID- 11485774 TI - Scleral infolding combined with vitrectomy and gas tamponade for retinal detachment with macular holes in highly myopic eyes. AB - PURPOSE: To describe the effectiveness of a surgical procedure, scleral infolding combined with vitrectomy and gas tamponade, for retinal detachment caused by macular holes in highly myopic eyes. METHODS: In a pilot study, scleral infolding was performed in 5 patients with macular holes, who were selected from 10 consecutive highly myopic patients with retinal detachment caused by macular holes (8 patients) or extramacular posterior-pole holes (2 patients), treated during 1 year at Okayama University Hospital. The patients were all women, 48-77 years of age (mean = 63.4 years), who had been followed-up for 1-2 years. Selection criteria for scleral infolding were either second surgeries for reopening of macular holes (2 patients) or residual retinal detachment around macular holes after complete fluid-air exchange with drainage of subretinal fluid at the initial surgery (3 patients). Following vitrectomy and complete epiretinal membrane removal in the posterior pole, the sclera was shortened by infolding on the temporal side. Three mattress sutures with 5-0 Dacron in each quadrant, 6 sutures in total, were placed at a 7-mm anteroposterior interval with posterior sutures located as deep as possible, near vortex veins. Fluid-gas exchange was then done, with or without endophotocoagulation applied around macular holes. RESULTS: After scleral infolding, macular holes were closed, and the retina was totally attached in all 5 patients. The final visual acuity ranged from 20/2000 to 20/70. CONCLUSION: Scleral infolding is a simple and effective procedure for treating retinal detachment with macular holes in highly myopic eyes and could be used as an optional procedure of reoperation for a failed initial vitrectomy. PMID- 11485775 TI - Surgical outcomes in juvenile retinal detachment. AB - PURPOSE: To evaluate retrospectively clinical features and surgical outcomes of rhegmatogenous retinal detachment in juvenile patients. METHODS: Between 1991 and 1996, 28 patients younger than 15 years of age with rhegmatogenous retinal detachment (32 eyes) underwent the first surgical procedure, scleral buckling and/or pars plana vitrectomy, at our hospital. RESULTS: The major types of juvenile detachment, in order of frequency, were idiopathic, familial exudative vitreoretinopathy, trauma, and high myopia. Proliferative vitreoretinopathy (PVR) of grade C or D was involved in 12 cases (37.5%). Among the 12 eyes with PVR, 7 attained retinal reattachment after the first surgery with scleral buckling. The overall reattachment rate was 28/32 (87.5%) after the first operation and 30/32 (93.8%) after the second operation. CONCLUSION: These findings indicate that the reattachment rate and visual prognosis can be as good in juvenile retinal detachment as in adult cases, when appropriate surgical procedures are used. PMID- 11485776 TI - A case of orbital solitary fibrous tumor. AB - BACKGROUND: Solitary fibrous tumor is a spindle cell neoplasm that most commonly arises in the pleura and very rarely involves the orbit. CASE: A 38-year-old woman presented with slowly progressive proptosis of 3 months duration and optic nerve head edema in her right eye. Magnetic resonance imaging revealed a well circumscribed, round mass lesion, which showed isointensity to the gray matter in a T1-weighted image, and variegated intensity in a T2-weighted image and contact with the optic nerve in her right orbit. The tumor was successfully removed by anterior orbitotomy. OBSERVATIONS: The tumor showed a "patternless pattern" of tumor cell arrangement, alternating hypercellular and hypocellular areas, a hemangiopericytoma-like pattern, and thickened strands of collagen. Immunohistochemically, the tumor cells were positive for CD34 and vimentin, and all were negative for other markers of epithelial, neural, muscular, histiocytic, and vascular endothelial cell elements. The tumor was diagnosed as a solitary fibrous tumor, and the patient was doing well with no evidence of recurrence 15 months after surgery. CONCLUSIONS: This case was the 19th reported case of solitary fibrous tumor in the orbital region. CD34 is a highly sensitive marker for solitary fibrous tumor. PMID- 11485777 TI - The Ishikawa classification of cavernous sinus lesions by clinico-anatomical findings. AB - PURPOSE: The Jefferson classification has been used to localize cavernous sinus lesions. However, this classification occasionally showed dissociation between identified localization and clinical findings. We investigated the clinical applicability of the newly proposed Ishikawa classification based on serial topographic sections of human cavernous sinus and the clinical findings. METHODS: In the Ishikawa classification, the cavernous sinus is divided into three portions, that is, anterior, middle, and posterior, demarcated by the location of the intracranial orifice of the optic canal and the entry of the maxillary nerve into the cavernous sinus. A total of 162 patients with cavernous sinus lesions were classified using both the Jefferson and the Ishikawa classifications and the clinical applicability of these two classifications was studied. Characteristics of the localization of lesions were also examined in each etiological type. RESULTS: By the Jefferson classification, 11% of the 162 patients had the anterior type of lesion, 12% the middle, 8% the posterior type, and 69% the unclassifiable type. However, by the Ishikawa classification, 35% had the anterior type, 10% the middle type, 22% the posterior type, 5% the whole type, and 28% the unclassifiable type of lesion. Furthermore, the Ishikawa classification revealed that the etiology of the anterior type was mainly inflammation, and that the etiology of the posterior and whole types was tumors. CONCLUSION: The Ishikawa classification is clinically useful to identify and classify the localization of cavernous sinus lesions. PMID- 11485778 TI - Optic neuropathy and central retinal vascular obstruction as initial manifestations of acute retinal necrosis. AB - BACKGROUND: The purpose of this brief communication is to alert ophthalmologists that optic neuropathy may herald acute retinal necrosis (ARN). CASE: A previously healthy 54-year-old man exhibited optic neuropathy as an initial presentation of ARN, 8 weeks after varicella-zoster dermatitis. OBSERVATIONS: Central retinal vascular obstruction developed subsequently in his left eye. Later, the classic presentation of ARN appeared in his contralateral eye. Systemic acyclovir therapy stopped the progression of retinitis and resulted in healing of retinal lesions in his right eye. CONCLUSIONS: This case suggests that optic neuropathy, especially with preceding herpetic dermatitis, should be suspected as the prodrome of ARN. PMID- 11485797 TI - Foot-and-mouth disease virus: cause of the recent crisis for the UK livestock industry. AB - The recent outbreak of foot-and-mouth disease (FMD) in the United Kingdom is a stark reminder of the economic devastation that this disease can wreak. Tracing the origin of such an outbreak is an essential part of disease control. Modern molecular methods have been in place for a number of years to enable scientists to identify unambiguously the strain of virus responsible. However, tracing the precise origin of such a strain is not so straightforward because the virus can move rapidly around the world with legal and illegal trade in animals and animal products. This short review describes the virus, its control and epidemiology. PMID- 11485798 TI - On the total number of genes and their length distribution in complete microbial genomes. AB - In sequenced microbial genomes, some of the annotated genes are actually not protein-coding genes, but rather open reading frames that occur by chance. Therefore, the number of annotated genes is higher than the actual number of genes for most of these microbes. Comparison of the length distribution of the annotated genes with the length distribution of those matching a known protein reveals that too many short genes are annotated in many genomes. Here we estimate the true number of protein-coding genes for sequenced genomes. Although it is often claimed that Escherichia coli has about 4300 genes, we show that it probably has only approximately 3800 genes, and that a similar discrepancy exists for almost all published genomes. PMID- 11485799 TI - Intrinsic errors in genome annotation. AB - Genome sequencing is usually followed by routine annotation of protein function based on the assumption that similar sequences will have similar functions. Here, we introduce a simple calculation to estimate the magnitude of any possible annotation errors. We counted the number of discrepancies in the annotation of well-established sets of similar proteins and extrapolated these values to the pairs of similar sequences used for the annotation of different microbial genomes. We conclude that the number of potential errors in the prediction of detailed functions is higher than is usually believed. PMID- 11485800 TI - Origin of an animal mitochondrial DNA polymerase subunit via lineage-specific acquisition of a glycyl-tRNA synthetase from bacteria of the Thermus-Deinococcus group. AB - Phylogenetic tree analysis shows that the accessory subunit animal mitochondrial DNA polymerase emerges as a result of horizontal transfer of the gene encoding glycyl-tRNA synthetase from a bacterium of the Thermus-Deinococcus group into the animal nuclear genome. This acquisition by a distinct eukaryotic lineage of a gene encoding a mitochondrial protein from a nonmitochondrial bacterial source underscores the contribution of different types of horizontal transfer event to the evolution of eukaryotes. PMID- 11485803 TI - RNAi surges on: application to cultured mammalian cells. PMID- 11485804 TI - Cytoplasmic incompatibility and maternal-haploid. PMID- 11485805 TI - When is a clone not a clone? PMID- 11485806 TI - Reversible regulation of mouse genes. PMID- 11485808 TI - Timing HIV infection. PMID- 11485809 TI - Never mind the quality... PMID- 11485810 TI - Common standards for genomics and proteomics. PMID- 11485811 TI - Plants to make silk. PMID- 11485812 TI - Copper transport meets development. PMID- 11485813 TI - Immunophenotyping leukemia by microarray. PMID- 11485814 TI - Genetic diversity and conservation efforts. PMID- 11485815 TI - Reawakening the telomeres. PMID- 11485816 TI - Transcriptional enhancers--on/off gene regulation as an adaptation to silencing in higher eukaryotic nuclei. AB - Transcriptional enhancers are DNA elements that act at a distance from a gene to regulate its expression. Principally, they seem to antagonize gene silencing, but they can also influence transcription rate. The relative importance and functional relationship of these two processes remains unclear. Cell differentiation requires a stable program of gene expression, with some genes active and others silent. Enhancers could function in this process by preventing incorporation of genes into heterochromatin, thus allowing the expression of genes characteristic of the cell lineage. This hypothesis takes into account the ability of enhancers to antagonize silencing, the association of silenced genes with heterochromatin in differentiated cells, and disruption of that association by enhancers. Comparison with yeast suggests that in higher eukaryotes, stronger and more numerous activating elements have been made necessary by the formation of a powerfully repressive nuclear compartment during cell differentiation. PMID- 11485817 TI - RNA silencing as a plant immune system against viruses. AB - "RNA silencing" refers to related processes of post-trancriptional control of gene expression found in plants, animals and fungi. A unifying feature of RNA silencing is that it mediates sequence-specific degradation of target transcripts, recruiting RNA molecules of 21-23 nucleotides as specificity determinants. In higher plants, RNA silencing serves as an adaptive, antiviral defence system, which is transmitted systemically in response to localized virus challenge. Plant viruses have elaborated a variety of counter-defensive measures to overcome the host silencing response. One of these strategies is to produce proteins that target the cell autonomous or signalling steps of RNA silencing. It is not known whether a similar antiviral mechanism also operates in animal cells. PMID- 11485819 TI - Intein spread and extinction in evolution. AB - Inteins are selfish DNA elements found within coding regions. They are translated with their host protein, but then catalyze their own excision and the formation of a peptide bond between their flanking protein regions. Understanding what drives and selects inteins is relevant for assessing whether they have unidentified biological functions and whether they can invade and become established in new genes and organisms. Inteins are suggested to have been present and more common in the progenitors of eukaryotes and prokaryotes. In these cells, inteins had some beneficial function or had evolved from an unknown beneficial protein. Since then, this putative benefit has been lost and inteins are gradually becoming extinct. The proteins in which inteins are currently found are proposed to be proteins vital for the survival of the organism, where intein removal is most difficult. PMID- 11485818 TI - MDM2: life without p53. AB - The MDM2 protein suppresses the ability of p53 to inhibit cellular proliferation or to induce cell death. This property underlies the oncogenic potential of MDM2, which is overexpressed in various human tumours. However, MDM2 also has p53 independent activities, which we focus on here. Similar to other oncogenes, surveillance pathways might counteract the deleterious effects of deregulated MDM2 expression. These pathways need to be inactivated for MDM2 oncogenic activity, which targets p53 but also other proteins. PMID- 11485820 TI - Preimplantation genetic diagnosis: the German situation. AB - Preimplantation genetic diagnosis makes it possible to detect some genetic disorders in embryos in vitro before they are transferred to the uterus. Using this technique, there is an opportunity for couples who have an increased risk of transmitting severe genetic disorders to their offspring to reduce this risk by >95%. By doing PGD, abortions at a later stage can be avoided. PMID- 11485821 TI - From controlling elements to transposons: Barbara McClintock and the Nobel Prize. PMID- 11485823 TI - Resveratrol and quercetin inhibit angiogenesis in vitro. AB - Resveratrol and quercetin are polyphenolic compounds found in grapes, red wine and other food products. In this study, we examined the effect of resveratrol and quercetin on the inhibition of angiogenesis in vitro. Resveratrol and quercetin inhibited the growth of bovine aorta endothelial (BAE) cells in a concentration dependent manner (6-100 microM).The migration of BAE was obviously inhibited by resveratrol and weakly inhibited by quercetin. When the lengths of all tubes constructed in the 3-D culture system with or without resveratrol or quercetin were measured, resveratrol was found to inhibit the tube formation of BAE cells in a concentration-dependent manner (6-100 microM). On the other hand, quercetin at concentrations above 100 microM significantly inhibited the tube formation of vascular endothelial cells. From these results, we suggest that resveratrol and quercetin may prove useful in the development of useful therapeutic agents or preventive food factors for tumor angiogenesis. PMID- 11485822 TI - Ras and Rho regulation of the cell cycle and oncogenesis. AB - The important contribution of aberrant Ras activation in oncogenesis is well established. Our knowledge of the signaling pathways that are regulated by Ras is considerable. However, the number of downstream effectors of Ras continues to increase and our understanding of the role of these effector signaling pathways in mediating oncogenesis is far from complete and continues to evolve. Similarly, our understanding of the components that control mitogen-stimulated cell cycle progression is also very advanced. Where our understanding has lagged has been the delineation of the mechanism by which Ras causes a deregulation of cell cycle progression to promote the uncontrolled proliferation of the cancer cell. In this review, we summarize our current knowledge of how deregulated Ras activation alters the function of cyclin D1, p21(Cip1), and p27(Kip1). The two themes that we have emphasized are the involvement of Rho small GTPases in cell cycle regulation and the cell-type differences in how Ras signaling interfaces with the cell cycle machinery. PMID- 11485824 TI - Pronounced inhibition by a natural anthocyanin, purple corn color, of 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-associated colorectal carcinogenesis in male F344 rats pretreated with 1,2-dimethylhydrazine. AB - The potential of purple corn color (PCC), a natural anthocyanin, to modify colorectal carcinogenesis was investigated in male F344/DuCrj rats, initially treated with 1,2-dimethylhydrazine (DMH), receiving 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP) in the diet. After DMH initiation, PCC was given at a dietary level of 5.0% in combination with 0.02% PhIP until week 36. No PCC-treatment-related changes in clinical signs, body weight and food consumption were found. Incidences and multiplicities of colorectal adenomas and carcinomas in rats initiated with DMH were clearly increased by PhIP. In contrast, lesion development was suppressed by PCC administration. Furthermore, in the non-DMH initiation groups, induction of aberrant crypt foci by PhIP tended to be decreased by the PCC supplementation. The results thus demonstrate that while PhIP clearly exerts promoting effects on DMH-induced colorectal carcinogenesis, these can be reduced by 5.0% PCC in the diet, under the present experimental conditions. PMID- 11485825 TI - Delayed effects of tamoxifen in hepatocarcinogenesis-resistant Fischer 344 rats as compared with susceptible strains. AB - The anti-oestrogenic drug tamoxifen has been under investigation as a breast cancer chemopreventive agent for at least a decade. However, its use for this purpose is still debatable since it is able to induce liver tumours in rats via a mechanism involving metabolic activation to a DNA adduct-forming electrophilic intermediate. The metabolic activation and adduct-forming properties of tamoxifen are now well characterized but less is known about its ability to induce hepatic cell proliferation, which is also essential for the carcinogenic process. The effects of tamoxifen on liver weight and cell proliferation were compared in female Fischer 344 (F344), Wistar and Lewis rats given the drug in the diet for up to 26 weeks. The onset and duration of hepatic cell proliferation varied between the strains of rat. In Wistar and Lewis but not F344 rats there was a marked increase in hepatocellular proliferation during the first 4 weeks of tamoxifen administration. In the Wistar strain this was associated with an increase in DNA adduct levels; no such increase was observed in the F344 strain. The onset of the proliferative response was delayed until the 13 week time point in the F344 strain. By the 13 and 26 week time points, cell proliferation in tamoxifen-treated Wistar and Lewis rat liver had returned to normal, but the amount of apoptotic activity in these livers was elevated. This suggests that excess cells generated during the proliferative phase of tamoxifen treatment were being eliminated by apoptosis. In the F344 strain, however, increased proliferative activity was associated with relatively low apoptotic activity at the 26 week time point, suggesting that the delayed proliferative response had yet to be balanced by apoptotic deletion. This is consistent with the fact that tamoxifen-induced hepatocellular tumours develop very late, towards the end of the lifespan, in this strain. The cell proliferative activity of tamoxifen in the Wistar rat liver was compared with that of a non-mutagenic analogue, toremifene. Tamoxifen induced increased cell cycle activity in the livers of rats following gavage dosing at all sampling times (1-12 weeks), whereas toremifene had no effect on the incidence of cycling in hepatic cells, demonstrating that the hepatic cell proliferation is not a general response to anti-oestrogen treatment. These observations suggest that the rate of promotion of liver tumours by tamoxifen is a function of the rate, time of onset and duration of increased cell replication. The susceptibility of rat strains to the hepatocarcinogenic effects of tamoxifen appears to depend upon the balance between initiation via DNA adduct formation, promotion via increased cell proliferation and cell deletion via apoptosis. Our findings suggest that an early proliferative response to tamoxifen is important in this process. PMID- 11485826 TI - Inhibition of growth of ES-2 human ovarian cancers by bombesin antagonist RC 3095, and luteinizing hormone-releasing hormone antagonist Cetrorelix. AB - We evaluated the effects of the bombesin/gastrin-releasing peptide (GRP) antagonist RC-3095, and the luteinizing hormone-releasing hormone (LH-RH) antagonist Cetrorelix, administered singly or in combination, on the growth of human ovarian carcinoma cell line ES-2, xenografted into nude mice. RC-3095 at a dose of 20 microg/day and Cetrorelix (100 microg/day), significantly reduced the volume of ES-2 tumors by 63.0% (P<0.01) and 38.0% (P<0.05) respectively, after 44 days of treatment, as compared with controls. The combination of RC-3095 with Cetrorelix inhibited the growth of ES-2 tumors by 66.2% (P<0.01). Serum levels of LH were significantly decreased in the groups treated with Cetrorelix alone and/or in combination with RC-3095. RT-PCR analyses revealed that the expression of mRNA for receptors of GRP (GRPR/BRS-1) and Neuromedin B (NMBR/BRS-2) on tumors was significantly decreased in all the treated groups. The expression of mRNA for epidermal growth factor receptors (EGFR) on tumors was reduced by 36.5 % (P<0.05) in the animals treated with Cetrorelix and by 72.5% (P<0.05) in the group that received the combination of RC-3095 with Cetrorelix. Our results indicate that the bombesin antagonist RC-3095 and the LH-RH antagonist Cetrorelix inhibit effectively the growth of ES-2 ovarian cancers in nude mice. These antagonists and their combination could be considered for the therapy of patients with ovarian cancer. PMID- 11485827 TI - tetra-O-methylnordihydroguaiaretic acid inhibits melanoma in vivo. AB - tetra-O-methylnordihydroguaiaretic acid is a derivative of a naturally-occurring lignan, nordihydroguaiaretic acid, that has previously been shown to inhibit various cancer types in vitro and in vivo. Additionally, nordihydroguaiaretic acid has been shown to have nephrotoxic effects in the rat. Here we show that tetra-O-methylnordihydroguaiaretic acid inhibits the growth of a number of tumor cell lines in vitro by inducing apoptosis in a non-schedule-dependent manner. Further, this compound inhibits the synthesis of DNA by melanoma cells and causes cell cycle arrest in G0/G1 and G2/M phases of the cell cycle. tetra-O Methylnordihydroguaiaretic acid also inhibits the growth of both murine and human melanomas and human colon cancer in vivo without apparent hepatic or renal toxicity. PMID- 11485828 TI - Tributyrin, an oral butyrate analogue, induces apoptosis through the activation of caspase-3. AB - The purpose of this study was to investigate the anti-proliferative and pro apoptotic effects of the butyrate analogues, tributyrin (TB) and phenylbutyrate (PB), in a colon cancer model. We demonstrate that HT-29 colon cancer cells exposed to PB and TB result in growth inhibition associated with an induction of apoptosis mediated through the activation of caspase-3 activity. A block in the G1/S cell cycle traverse associated with a decrease in CDK2 (cyclin dependent kinase) protein levels and retinoblastoma protein hypophosphorylation was also noted after PB and TB exposure. Importantly, TB proved to be the most potent agent in its ability to induce these phenotypic changes, and potentially may represent a novel therapy for patients with advanced colorectal cancer. PMID- 11485829 TI - Expression of multiple larger-sized transcripts for several genes in oligodendrogliomas: potential markers for glioma subtype. AB - Astrocytomas and oligodendrogliomas are two brain tumors that follow different clinical courses. Although many of these tumors can be identified based on standard histopathological criteria, a significant percentage present notable problems in diagnosis. To identify markers that might prove useful in distinguishing glioma subtypes, we prepared and analyzed cDNA libraries for differential expression of genes in an astrocytoma (grade II), an oligodendroglioma (grade II), and a meningioma (benign). The tumor libraries were compared by sequencing randomly selected clones and tabulating the expression frequency of each gene. In addition to identifying several genes previously reported or expected to be differentially expressed among these tumors, several potential new brain tumor markers were identified and confirmed by Northern blot analysis of a panel of brain tumors. A surprising result of this analysis was the observation that several larger-sized transcripts for various genes were predominantly expressed in the oligodendroglioma tumors, when compared to the other brain tumors or in non-tumor gray matter. These findings are consistent with different pre-mRNA splicing patterns observed between oligodendrogliomas and astrocytomas. In support of this hypothesis, our screen revealed significantly higher levels of two hnRNP A1 transcripts in oligodendrogliomas. hnRNP A1 is a component of the spliceosome whose expression levels affect splice site selection in vivo. The preferential expression of larger-sized transcripts for several genes in oligodendrogliomas may be useful for distinguishing astrocytic and oligodendroglial gliomas. PMID- 11485830 TI - The activation of PI 3-K and PKC zeta in PMA-induced differentiation of HL-60 cells. AB - The human myelocytic leukemia cell line HL-60 is a useful model for the study of cellular differentiation. Phorbol 12-myristate 13-acetate (PMA) induces the monocyte/macrophage-like differentiation of HL-60 cells and results in growth arrest, increasing adherence. In PMA-induced differentiation of HL-60 cells, phosphoinositide 3-kinase (PI 3-K) activity was measured as phosphatidylinositol3P recovery from phosphatidylinositol by in vitro kinase assay. PI 3-K activity was increased in HL-60 cells that were stimulated by 20 nM PMA and the activity was inhibited by pretreatment with 20 microM LY294002, a specific inhibitor of PI 3-K. Members of the protein kinase C (PKC) family have been suggested to be one of the downstream targets of PI 3-K. PKC zeta is one of the atypical PKCs, non-diacylglycerol-responsive PKCs, and the activity was measured by the ability of phosphorylation onto myelin basic protein. PMA also induced the activation of PKC zeta during monocytic differentiation of HL-60 cells, and LY294002-pretreated cells failed to induce PKC zeta activation. The activity of PI 3-K is essential for PKC zeta activation, and LY294002 blocks both monocytic differentiation of HL-60 cells and activation of PKC zeta during PMA induced cell differentiation. This implies that activated PI 3-K subsequently stimulates the PKC zeta in the process of PMA-induced monocytic differentiation. PMID- 11485831 TI - B-myb rescues ras-induced premature senescence, which requires its transactivation domain. AB - B-myb, a ubiquitously expressed member of the myb gene family, is highly regulated throughout the cell cycle and appears to be required for cell cycle progression. In contrast to its relatives A-myb, c-myb, and v-myb, no transforming activity of B-myb has been reported thus far. We report here that B myb can rescue senescence induced by an activated ras oncogene in rodent cells in vitro. We show that transformation by B-Myb involves its ability to activate transcription. Similar to other oncogenic transcription factors, such as c-Myc and E2F, we show that B-Myb also has repression activity. We demonstrate that the C-terminus of B-Myb can function as a repressor of transcription, that B-Myb interacts with the repressor molecules BS69 and N-CoR and that the repression function, like the transactivation domain, contributes to B-myb transformation. PMID- 11485832 TI - Association of maspin expression with the malignancy grade and tumor vascularization in breast cancer tissues. AB - Maspin belongs to a tumor suppressing protein which is usually expressed highly in myoepithelial cells, and is significantly downregulated in breast cancer cells. We focused on identifying the correlation between maspin expression and the clinicopathological features of human breast cancer tissues using immunohistochemistry. There was a significant correlation in that maspin-positive tumor specimens showed a low pathological grade of malignancy, such as a lower infiltration of the tumor into the surrounding tissue and a downregulation of c erbB2 expression. Moreover, maspin-positive cases showed a significant decrease in tumor vessels positively stained with anti-factor VIII-related antigen antibody. These results suggest that maspin production in myoepithelial cells could downregulate the malignant phenotype of breast cancer. PMID- 11485833 TI - CYP1A2 is expressed along with CYP1A1 in the human lung. AB - The expression and activity of CYP1A1 were examined in fresh, small-sized lung biopsy specimens from nine human subjects. CYP1A1 transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR) analysis of total lung RNA. CYP1A2 transcripts were detected in the RNA samples as well, and bioactivation of 2-aminofluorene (2-AF) or 2-amino-3,4-dimethylimidazo[4,5 f]quinoline (MeIQ), a CYP1A2-preferential activity, was catalyzed by the lung S(9) fractions also. Two major bands were detected in the whole homogenate by western blot analysis using CD3, a mouse anti rat CYP1A1 monoclonal that cross reacts with rat CYP1A2 as well as with human CYP1A1 and CYP1A2. S(9) fractions from the tissues catalyzed the bioactivation of benzo[a]pyrene (B[a]P), a CYP1A1 preferential activity, to mutagens in the Ames assay. Our findings are in agreement with the known presence of CYP1A1 in the human lung, and provide strong evidence for the expression of catalytically functional CYP1A2 in the tissue. PMID- 11485834 TI - Neurobehavioral deficits associated with PCB in 7-year-old children prenatally exposed to seafood neurotoxicants. AB - Prenatal exposure to polychlorinated biphenyls (PCBs) was examined by analysis of cord tissue from 435 children from a Faroese birth cohort. Analysis of 50 paired cord blood samples showed excellent correlation with the cord tissue concentration (r=.90). Among 17 neuropsychological outcomes determined at age 7 years, the cord PCB concentration was associated with deficits on the Boston Naming Test (without cues, two-tailed P=.09 not adjusted for mercury; with cues, P=.03), the Continuous Performance Test reaction time (P=.03), and, possibly, on long-term recall on the California Verbal Learning Test (P=.15). The association between cord PCB and cord-blood mercury (r=.42) suggested possible confounding. While no PCB effects were apparent in children with low mercury exposure, PCB associated deficits within the highest tertile of mercury exposure indicated a possible interaction between the two neurotoxicants. PCB-associated increased thresholds were seen at two of eight frequencies on audiometry, but only on the left side, and no deficits occurred on evoked potentials or contrast sensitivity. The limited PCB-related neurotoxicity in this cohort appears to be affected by concomitant methylmercury exposure. PMID- 11485835 TI - Differential effects of two NMDA receptor antagonists on cognitive-behavioral development in nonhuman primates I. AB - The present experiment examined effects of chronic exposure to remacemide (an N methyl-D-aspartate [NMDA] antagonist which also blocks fast sodium channels) or MK-801 (which blocks NMDA receptors, exclusively) on learning and motivation in young rhesus monkeys. Remacemide (20 or 50 mg/kg/day) or MK-801 (0.1 or 1.0 mg/kg/day) was administered every day to separate groups of animals via orogastric gavage for up to 2 years. Immediately prior to dosing, 5 days per week (M--F), throughout the 2-year dosing period, an incremental repeated acquisition (IRA) task was used to assess learning and a progressive ratio (PR) task was used to assess motivation. The results indicate an effect of 50 mg/kg/day remacemide to impair learning (IRA) which persisted even after drug treatment was discontinued. MK-801 had no effect on learning but transiently increased motivation. Because the effects of remacemide occurred independently of changes in motivation or response rates, they are likely due to specific cognitive impairments and are not due to an inability of subjects to fulfill the motoric requirements of the task. The fact that MK-801 did not alter learning suggests that NMDA antagonism alone may be insufficient to produce learning deficits in young monkeys and that such deficits may rely on the ancillary blockade of fast sodium channels. PMID- 11485837 TI - Methamphetamine concentrations in fetal and maternal brain following prenatal exposure. AB - Levels of methamphetamine in maternal striatum and whole fetal mouse brain were assessed at 0.5, 1, 2, and 4 h postinjection on gestational day 14 (GD14) following a single, subcutaneous injection of 40 mg/kg (+)-methamphetamine hydrochloride to pregnant mice. In the dams, striatal concentrations of methamphetamine peaked at 1 h postinjection, reaching levels of approximately 510 ng/mg protein. Amphetamine, the primary metabolite of methamphetamine, increased to 77 ng/mg protein at 2 h and remained elevated by 4 h postinjection. In the fetal brain, peak methamphetamine concentrations of approximately 122 ng/mg protein were attained at 1 h. Amphetamine was only detectable in fetal brain at 2 and 4 h postinjection. Regional analysis of methamphetamine levels in fetal striatum, cortex, and brainstem revealed that the drug was not uniformly distributed. Maternal administration of methamphetamine results in fetal brain drug concentrations, which approximate those reported in human infants whose mother abused methamphetamine. This dosage regimen, therefore, serves as an appropriate animal model for assessing the potential risks to human offspring exposed to methamphetamine in utero. PMID- 11485836 TI - Differential effects of two NMDA receptor antagonists on cognitive--behavioral performance in young nonhuman primates II. AB - The present experiment examined the effects of chronic exposure to remacemide (an NMDA antagonist that also blocks fast sodium channels) or MK-801 (which blocks NMDA receptors more selectively) on the acquisition of color and position discrimination and short-term memory behavior in juvenile rhesus monkeys. Throughout the 2-year dosing period, a conditioned position responding (CPR) task was used to assess color and position discrimination and a delayed matching-to sample (DMTS) task was used to assess memory. Chronic exposure to high doses of either drug delayed the acquisition of accurate color and position discrimination without altering response rates. In the case of MK-801, these effects abated within 6 months of the start of treatment. In the case of remacemide, the effects persisted for 17 months of dosing. Neither compound significantly altered performance of the short-term memory task at any time point or at any dose tested. The fact that the effects of remacemide on behavioral performance were more persistent than those seen for MK-801 suggests that tolerance may develop to the behavioral effects of MK-801, which does not develop to the effects of remacemide. Alternatively, these results may suggest that the concurrent antagonism of NMDA receptors and fast sodium channels may have more profound consequences for behavior than does the antagonism of NMDA receptors alone. PMID- 11485838 TI - Effect of prenatal ethanol exposure during the brain growth spurt of the guinea pig. AB - This study tested the hypothesis that prenatal ethanol exposure during the last third of gestation, including the brain growth spurt (BGS), in the guinea pig produces neurobehavioural teratogenicity, manifesting as brain growth restriction and hyperactivity. Pregnant guinea pigs (term, about gestational day (GD) 68) received oral administration of ethanol (2 g/kg maternal body weight per day on GD 43 and/or GD 44 and then 4 g/kg maternal body weight per day from GD 45 to GD 62), isocaloric-sucrose/pair-feeding, or water. Maternal blood ethanol concentration (BEC) on GD 57 or 58, at 1 h after the daily dose, was 340+/-76 mg/dl (n=8). Ethanol treatment decreased brain, cerebral cortical, hippocampal, and cerebellar weights at GD 63 (P<0.05), and decreased brain and cerebral cortical weights at postnatal day 10 (P<0.05), with no effect on body weight and no apparent effect on spontaneous locomotor activity. The data demonstrate that, in the guinea pig, prenatal ethanol exposure during the last third of gestation, including the BGS, decreases brain weight that persists into postnatal life, which is associated with growth restriction of the cerebral cortex. However, this prenatal ethanol exposure regimen, including the BGS, does not increase spontaneous locomotor activity in contrast to the persistent hyperactivity that occurs after chronic ethanol exposure throughout gestation. PMID- 11485839 TI - Long-term consequences of developmental exposure to aluminum in a suboptimal diet for growth and behavior of Swiss Webster mice. AB - Swiss Webster mice received diets containing 7 (control), 100, 500, or 1000 microg aluminum (Al)/g throughout development (conception to 35 days of age) and were tested behaviorally as adults (>90 days of age). The basal diet contained the same percent of recommended dietary amounts of phosphate, calcium, iron, magnesium, and zinc as young women usually consume. These "realistic" dietary conditions led to 12--15% growth retardation in the Al1000 group at the time of testing. Females were evaluated in a cognitive task (Morris water maze) at 3 months of age and males were evaluated in a motor test battery at 5 months of age. Al1000 females (n=16) were slower than controls in learning the Morris maze, as suggested by fewer mice with low latencies during the first three sessions of the four-session learning series. Influences of Al on cue utilization were also found in probe sessions eliminating salient or nonsalient cues. With motor testing, the Al1000 males (n=20) had significantly lower hindlimb grip strength than controls, an effect that was eliminated by covariance analysis with body weight. Subtle influences of Al on rotarod and wire suspension tests were also noted. The data suggest that developmental Al exposure under normal, but less than optimal, dietary conditions can lead to subtle but long-term effects on growth and brain function in adulthood. PMID- 11485840 TI - Maternal urinary tract infection alters water maze performance in the offspring. AB - The effects of maternal urinary tract infection (UTI) or endotoxin exposure on fetal outcome in rats were investigated. Prior to conception, dams of the UTI group were water-deprived and anesthetized. The urinary tract was then catheterized and injected with 0.2 of 1 x 10(9) Escherichia coli. The endotoxin group was injected with 0.03 mg/kg lipopolysaccharide on the fourth day of gestation and then every third day thereafter. The control groups were treated in the same manner, with the exception that the infection control was not catheterized or injected with E. coli, and the endotoxin control was not exposed to lipopolysaccharide. A nontreated control group was weighed daily. Beginning on postnatal day (PD) 19, offspring were tested daily in a water maze spatial navigation task. The retention latencies (Sessions 7--10) revealed deficits in the infection and endotoxin groups. In the rat model, these findings suggest that exposure during gestation to a maternal immune challenge may result in adverse fetal outcome. PMID- 11485841 TI - Evaluation of acute sensory--motor effects and test sensitivity using termiticide workers exposed to chlorpyrifos. AB - Sensory and motor testing was performed on a group of termiticide workers primarily using chlorpyrifos-containing products to evaluate both the acute effects from current exposure and sensitivity of the measures to detect effects. The study group comprised 106 applicators and 52 nonexposed participants. Current exposure was measured by urinary concentrations of 3,5,6-trichloro-2-pyridinol (TCP) collected the morning of testing. The mean TCP value for the 106 applicators was 200 microg/g creatinine. Participants received 4--5 h of testing and were evaluated using a sensory--motor test battery recommended by a National Institute for Occupational Safety and Health (NIOSH)-sponsored advisory panel to be appropriate for testing effects from pesticide exposures. Measurements testing olfactory dysfunction, visual acuity, contrast sensitivity, color vision, vibrotactile sensitivity, tremor, manual dexterity, eye--hand coordination, and postural stability were analyzed. Study results indicated limited acute effects from exposure to chlorpyrifos using urinary TCP as a measure of current exposure. The effects occurred primarily on measures of postural sway in the eyes closed and soft-surface conditions, which suggests a possible subclinical effect involving the proprioceptive and vestibular systems. Several other tests of motor and sensory functions did not show any evidence of acute exposure effects, although statistically significant effects of urinary TCP on the Lanthony color vision test scores and one contrast sensitivity test score were found. The visual measures, however, were not significant when a step-down Bonferroni correction was applied. Information also is presented on the sensitivity of the measures to detect effects in an occupationally exposed population using standard error of the parameter estimates. PMID- 11485842 TI - Localization of tetra(m-hydroxyphenyl)chlorin (Foscan) in human healthy tissues and squamous cell carcinomas of the upper aero-digestive tract, the esophagus and the bronchi: a fluorescence microscopy study. AB - To date, little is known about precise time-dependent distribution and histological localization of tetra(m-hydroxyphenyl)chlorin (mTHPC) in human healthy tissues and squamous cell malignancies in the upper aero-digestive tract. A fluorescence microscopy study was performed on 50 healthy tissue biopsies and on 13 tumors (graded from Tis to T1 SCC) from 30 patients. Tissue samples were taken between 4 h and 11 days following injection of 0.15 mg/kg mTHPC. A fairly comparable distribution pattern in various tissues was observed over time in different patients. Vascular localization of mTHPC fluorescence predominates at a short delay, whereas the dye is essentially located in the tumoral and healthy mucosa after longer delays. A much lower uptake and retention of mTHPC fluorescence was noted in striated muscle and cartilage as compared to neoplastic lesions. No significant selectivity was found between healthy and tumoral mucosa. The obtained data are important to confirm drug-light interval that have been selected for effective PDT for early SCC malignancies while minimizing the risks of over- or under-treatment. The low fluorescence level in striated muscle provides the opportunity to develop interstitial PDT as a treatment modality for invasive SCC of unfavorable locations in the oral cavity or pharynx, such as the base of the tongue. PMID- 11485843 TI - Photosensitizer initiated attacks on DNA under dry conditions and their inhibition: a DNA archiving issue. AB - Long-term aging of dry DNA is thought to be due to the attack of diverse cascades of reactive species with probably, no one single initiator of the cascades explaining all circumstances. Photosensitizer-initiated reactions from methylene blue and riboflavin were used to generate two model systems of reactive species around dry DNA in order to understand such systems and how to block them. Damage was assessed using plasmid DNA as a substrate with an in-situ microgel electrophoretic technique. Photodynamic methylene blue damage to DNA was very oxygen dependent but not that of riboflavin. This indicates that indirect type II pathways, probably via singlet oxygen were important for methylene blue but not for riboflavin. In both the absence and presence of oxygen, the DNA protection offered by dry caffeine and urate to both photodynamic agents indicated that most DNA attack was via electrophilic species. Overall, protection of dry archived DNA from spontaneously reactive species such as free radicals appears to be a real issue and, as expected, the predominant species in air appear to involve oxygen but not exclusively or necessarily so. PMID- 11485844 TI - In vivo pharmacokinetics of protoporphyrin IX accumulation following intracutaneous injection of 5-aminolevulinic acid. AB - Photodynamic therapy with 5-aminolevulinic acid (ALA) derived protoporphyrin IX (PpIX) as photosensitizer is a promising treatment for basal cell carcinomas. Until now ALA has been administered topically as an oil-in-water cream in most investigations. The disadvantage of this administration route is insufficient penetration in deeper, nodular tumours. Therefore we investigated intracutaneous injection of ALA as an alternative administration route. ALA was administered in 6-fold in the normal skin of three 6-week-old female Dutch pigs by intracutaneous injection of an aqueous solution of ALA (pH 5.0) in volumes of 0.1-0.5 ml and concentrations of 0.5-2% and by topical administration of a 20% ALA cream. During 8 h fluorescence of ALA derived PpIX was measured under 405 nm excitation. For the injection the measured fluorescence was shown to be dose dependent. All injected doses of 3 mg ALA or more lead to a faster initial increase rate of PpIX synthesis and significantly greater fluorescence than that measured after topical administration of ALA. Irradiation (60 Jcm(-2) for 10 min) of the spots was performed at 3.5 h after ALA administration. After 48 and 96 h visual damage scores were evaluated and biopsies were taken for histopathological examination. After injection of 2 mg ALA or more the PDT damage after illumination was shown to be significantly greater than after topical application of 20% ALA. An injected dose of 10 mg ALA (0.5 ml of a 2% solution) resulted in significantly more tissue damage after illumination than all other injected doses. PMID- 11485845 TI - Photodynamic therapy with topical delta-aminolaevulinic acid for the treatment of plantar warts. AB - Treatments currently employed for plantar warts are often painful (electrosurgery, cryotherapy) and not always effective (keratolytic agents). In this paper we investigate the effect of photodynamic therapy (PDT) with topical delta-aminolaevulinic acid (ALA) on plantar warts. In order to remove the superficial hyperkeratotic layer of the warts an ointment containing 10% urea and 10% salicylic acid was applied for 7 days. After gentle curettage, a cream containing 20% ALA was applied under an occlusive dressing for 5 h on 64 warts, while 57 warts (controls) received only the vehicle. Both the ALA-treated warts and the controls were irradiated using a visible light lamp (with a range of 400 700 nm, peaking at 630 nm). The light dose was 50 J/cm(2). Patients were followed up for 22 months. Two months after the last irradiation session 48 (75.0%) out of 64 ALA-PDT treated warts had resolved. By contrast only 13 (22.8%) of the 57 control warts had done so. During the treatment a few patients complained of a mild burning sensation. The absorption of ALA by the verrucous tissue was demonstrated by in vivo fluorescence spectroscopy. This study shows that topical ALA-PDT can be an alternative treatment for plantar warts. Further studies will be necessary in order to optimize the concentration of ALA and duration of treatment. PMID- 11485846 TI - Photodegradation of natural organic matter exposed to fluctuating levels of solar radiation. AB - Irradiation of natural water samples with natural or artificial UVR typically results in a progressive loss of color and decreased absorbance; a process often referred to as photobleaching. In a typical photobleaching experiment, samples are exposed to a relatively constant level of artificial or natural UVR. However, under most natural situations, the vertical mixing of the water within the upper mixed layer results in strong and periodic fluctuations in UV irradiance. In this paper, we present the results of an experiment in which natural lake water was exposed to solar radiation in quartz tubes that were incubated either at fixed depths or rotating within the water column. We found differences between rotating and fixed samples in (i) photobleaching, (ii) nutrient release, and (iii) subsequent use by algae and bacteria. The evidence presented in this study demonstrated that photochemical processes might be affected by vertical water motion. The reasons for such differences remain largely unknown. Although we offer a potential explanation for such differences, our proposed mechanism is based on a post-hoc analysis of the data and should be taken solely as a working hypothesis for future research. PMID- 11485847 TI - Galactolipids not associated with the photosynthetic apparatus in phosphate deprived plants. AB - The galactolipid digalactosyldiacylglycerol (DGGD) is one of the major constituents of thylakoids, accounting for about 25% of polar lipids found in these membranes. Although the presence of DGDG has frequently been correlated with the structural and functional integrity of the photosynthetic apparatus, it is still a matter of debate of what the in-vivo function of DGDG actually might be. To further the understanding of the role of DGDG within the photosynthetic apparatus, experiments were conducted on different Arabidopsis thaliana lines with altered DGDG content. The dgd1 mutant is characterized by a 90% reduction in the DGDG content, resulting in a severe dwarfism during growth. Complementation of the dgd1 mutant with a DGD1 cDNA completely restored the wild-type characteristics, while photosynthesis-related parameters were intermediate in transgenic plants with a partial reduction in DGD1 activity caused by post transcription gene silencing due to over-expression of a DGD1 cDNA in wild-type plants. These data provide clear evidence for a causal relationship between the DGDG content, and the structure and function of the photosynthetic apparatus. However, a significant DGDG accumulation in the dgd1/pho1 double mutant was without any detectable effect on photosynthetic activity, indicating that the molecular DGDG species synthesized upon phosphate deprivation in leaves cannot substitute for the DGDG species present under normal nutrient supply of plants. It is suggested that depending on the environmental growth conditions different pools of DGDG species exist in plants of which one is not associated with the photosynthetic apparatus. PMID- 11485848 TI - New vegetation indices for remote measurement of chlorophylls based on leaf directional reflectance spectra. AB - Directional reflectance (R) spectra from 380 to 780 nm for nadir illuminated leaves of four different plants (croton, Codiaeum variegatum; spotted eleagnus, Eleagnus pungens Maculata; Japanese pittosporum, Pittosporum tobira and Benjamin fig, Ficus benjamina Starlight) were acquired at a viewing angle of 30 degrees from the nadir direction. Chlorophyll-a and -b content of leaves covered a range of 1-60 and 0.5-21 microg/cm(2), respectively. In contrast with previous results from hemispherical reflectance measurements, directional reflectance data does not correlate well with chlorophyll concentration. This is mainly due to the external reflectance (R(E)) at the leaf epidermis, caused by the mismatch of the refractive index at the air-epidermis and epidermis-inner layer boundary. The external reflectance can be identified with the blue flat reflectance between 380 and 480 nm. The inner reflectance (R(I)), obtained by subtracting the external reflectance from the measured spectra, was found to be linearly related to the logarithm of the chlorophyll content. Good fitting of the log (Chl) versus R(I)(lambda) curves were obtained for R(I) in the green band (around 550 nm) and close to the inflection point in the red edge (around 700 nm). The coefficient of determination, r(2), of curve fitting improved (up to 0.97) when the normalised inner reflectance NR(I)(lambda)=R(I)(lambda)/R(I)(lambda(0)), with lambda(0)>or=750 nm, was used instead of the absolute reflectance. The best indices for Chl, Chl-a and Chl-b determination were R(I)(542)/R(I)(750), R(I)(706)/R(I)(750) and R(I)(556)/R(I)(750), respectively. However, since the content of Chl-a relative to Chl-b was almost constant for the plants investigated, the two last indices must be further validated on leaves with a high variability in the Chl-a:Chl-b ratio. The error in the determination of chlorophyll content was found to be of the order of 10%. This value was lower than those obtained by applying the vegetation indices previously suggested. Therefore, the normalised inner reflectance in the green and in the red edge represents a more suitable index for the chlorophyll determination than those up to now used. PMID- 11485849 TI - Scavenging of acetylperoxyl radicals and quenching of triplet diacetyl by beta carotene: mechanisms and kinetics. AB - Beta-carotene scavenges triplet diacetyl generated by laser flash photolysis with a second-order rate constant of 9.1+/-0.9 x 10(9) M(-1) s(-1) in deaerated benzene at 20 degrees C. In the presence of oxygen diacetyl dissociates to generate acetylperoxyl radicals. It is demonstrated that diacetyl does not dissociate to any appreciable extent in the absence of oxygen. The acetylperoxyl radical is scavenged by beta-carotene with second-order rate constant 9.2+/-0.6 x 10(8) M(-1) s(-1) in aerated benzene at 20 degrees C to give an adduct between the acetylperoxyl radical and beta-carotene, whereas no evidence of oxidation of beta-carotene by the strongly oxidizing acetylperoxyl radical to give the beta carotene radical cation is found. This adduct decays with first-order rate constant 1.35+/-0.16 x 10(3) s(-1) to give (presumably) a beta-carotene epoxide and the acetyloxyl radical. PMID- 11485850 TI - Long-lived reactive intermediate photogenerated from N-(5-azido-2-nitrobenzoyl) N'-(D-biotinyl)-1,2-diaminoethane as an affinity reagent to streptavidin. AB - Irradiation of a complex between N-(5-azido-2-nitrobenzoyl)-N'-(D-biotinyl)-1,2 diaminoethane (I) and streptavidin with light of 313 nm led to the covalent attachment of the photobiotin analogue I to the protein. Streptavidin could also be labelled in the dark with prephotolyzed I. These results indicate that a long lived reactive intermediate was formed upon irradiation. Moreover, after cleavage of labelled streptavidin with proteinase K this intermediate appears to be covalently attached to the same peptide as the one obtained by direct photoaffinity labelling. An iminosulfurane II derived from the reaction of biotin sulfur atom with aryl nitrene is responsible for the dark-labelling reaction. The photoproduct II converts in an aqueous solution almost completely into N-(5-amino 2-nitrobenzoyl)-N'-(D-(S-oxo)biotinyl)-1,2-diaminoethane (the half-life of II is 10 days). PMID- 11485851 TI - Fluorescence spectroscopy of normal mouse skin exposed to 5-aminolaevulinic acid and red light. AB - Photobleaching and phototransformation of protoporphyrin IX (PpIX) was investigated in normal mouse skin. The PpIX was induced by topical application of 5-aminolaevulinic acid (ALA). Exposure to laser light (635 nm) caused photobleaching of PpIX fluorescence and formation of fluorescent products. Analysis of the fluorescence spectra revealed appearance of new fluorescent photoproducts during light exposure. The main photoproduct, supposedly chlorin type photoprotoporphyrin (PPp), exhibited fluorescence with an emission maximum at 675 nm. The other products exhibited main fluorescence peaks at around 588 and 623 nm that can presumably be attributed to an endogenous metallo-porphyrin and water-soluble porphyrin(s), respectively. Our results indicate that light exposure causes alterations in the enzymatic pathway of PpIX synthesis from ALA and leads to accumulation of intermediate water-soluble porphyrins. ALA-induced porphyrins are transported away from the treated area and partly deposited in remote skin sites. PMID- 11485853 TI - Ramzi S. Cotran, M.D., 1932-2000. PMID- 11485854 TI - Arrhythmogenic right ventricular cardiomyopathy: An update. PMID- 11485855 TI - Cell proliferation in the growing human heart: MIB-1 immunostaining in preterm and term infants at autopsy. AB - Few studies of human cardiac myocyte proliferation in the perinatal period have been conducted. We measured the proliferative activity of left ventricular myocytes in tissue obtained at autopsy in three surgically induced abortuses, 20 preterm infants with gestational ages ranging from 12 to 35 weeks, eight term infants with ages ranging from 1 day to 11 months, and five adults. The preterm infants lived less than 24 h, thus simulating the in utero condition of developing hearts. To assess the proliferative activity of the myocytes, we measured immunoreactivity using the monoclonal antibody MIB-1 against the recombinant Ki-67 nuclear antigen. Immunostained sections were examined by light microscopy, and the results expressed as a staining index (SI) of 0-3, according to the percentage of positively stained myocyte nuclei. Myocyte proliferative activity remained constant during the early preterm period and decreased in the late preterm and early postterm periods. Adult myocytes, regardless of cardiac weight, did not reveal proliferative activity as assessed by immunostaining. This proliferation pattern is consistent with findings in most earlier studies in animal models. PMID- 11485856 TI - Interaction between sarcomere and mitochondrial length in normoxic and hypoxic rat ventricular papillary muscles. AB - We hypothesized that the mitochondrial length may be altered according to changes in the sarcomere length, and that this relationship may be affected by exposure to hypoxia. Rat ventricular papillary muscles were isolated and immersed in normoxic or hypoxic solutions for 10 min. Sarcomeres of various lengths were obtained by fixing the papillary muscles in a slack or stretched state, or after exposure to a contracture solution containing saponin and CaCl(2). The mitochondrial length measured using electron microscopy significantly correlated to the length of the adjacent sarcomere in both the normoxic (n=767) and hypoxic (n=1145) groups (P<.0001). The slope of the regression line, however, was significantly less steep, and its intercept was significantly larger in the hypoxic group than in the normoxic group (analysis of covariance). When we analyzed the mitochondrial lengths among the three sarcomere-length subgroups (<1.5, 1.5-2.0, and >2.0 microm), the mitochondrial length was significantly shorter in the hypoxic condition than in the normoxic condition at sarcomere lengths greater than 2.0 microm. Staining for desmin, the major muscle-type intermediate filament, the longitudinal system of which connects the mitochondria with the Z bands of sarcomeres, showed a clear cross-striation pattern in both papillary muscles with and without the exposure to hypoxia, suggesting that desmin was preserved after the exposure to hypoxia. These data indicate that the mitochondrial length changes according to changes in the sarcomere length, suggesting the possible role of mitochondria as an internal load against myocyte contraction. It is also suggested that mitochondria exposed to hypoxia may be more resistive to both compression and stretch in a longitudinal direction than those in the normoxic condition. PMID- 11485857 TI - Smooth muscle cell depletion and collagen types in progeric arteries. AB - Review of two autopsy cases of progeria confirms severe smooth muscle cell (SMC) depletion in the atherosclerotic aortic media and the presence of collagen types I, III, IV, V, and VI in the aorta and renal vessels as is consistent with atherosclerotic disease. PMID- 11485858 TI - Primary extramedullary plasmacytoma in the atria of the heart. AB - A rare case of primary extramedullary plasmacytoma of the heart is described. The large space-occupying tumor in the atrial wall and its associated significant pericardial effusion caused marked impairment of cardiovascular function. The diagnosis was made via intravenous biopsy. Immunohistochemistry demonstrated CD45 and kappa light chain expression. Other B-cell- and T-cell-specific lymphoid markers were negative. Elevated kappa light chain was detected in serum by immunofixation electrophoresis. The differential diagnosis of plasmacytoma should be considered in cardiac tumors when routine screening is negative with lymphoid (CD20, CD3) and soft tissue immunohistochemical markers. PMID- 11485859 TI - Spontaneous coronary artery dissection associated with cocaine use: a case report and brief review. AB - Isolated, spontaneous dissection of the coronary arteries in the absence of trauma is an unusual but well-documented occurrence. Fewer than 50 cases have been reported in males in the English language literature, and only one case, nonfatal, was associated with cocaine use. We present the second overall and the first fatal case of cocaine-associated spontaneous coronary artery dissection and a brief review of the literature on coronary dissection and the cardiovascular effects of cocaine use. The mechanism of cocaine's toxicity on the heart and vasculature is complex, multifactorial, and predominantly related to cocaine's adrenergic properties. The increased arterial blood pressure from cocaine's inotropic and chronotropic effects combined with its direct vasoconstrictive effect leads to increased shear forces on the coronary endothelium. This elevated stress may be responsible for the formation of an intimal tear and the subsequent dissection of the coronary artery. If the dissected portion of the arterial wall is displaced enough to significantly occlude the true lumen, infarction can result. In light of this possibility, coronary artery dissection must be considered in young patients presenting with symptoms of cardiac ischemia and a history of cocaine use. PMID- 11485860 TI - Primary pericardial mesothelioma. AB - Primary pericardial mesothelioma is an extremely rare tumor. This case illustrates the typical late presentation of primary pericardial mesothelioma with symptoms and signs of constrictive pericarditis. An unusual feature was the complete encasement of the heart by the tumor. No satisfactory treatment was available. PMID- 11485861 TI - Marine swimming-related illness: implications for monitoring and environmental policy. AB - There is increasing evidence that environmental degradation may be contributing to an increase in marine-related diseases across a wide range of taxonomic groups. This includes a growing number of reports of both recreational and occupational users of marine waters developing gastrointestinal, respiratory, dermatologic, and ear, nose, and throat infections. The duration and type of exposure, concentration of pathogens, and host immunity determine the risk of infection. Public health authorities may not be able to accurately predict the risk of waterborne disease from marine waters due to the limitations of conventional monitoring, as well as erroneous perceptions of pathogen life span in marine systems. Pathogens undetectable by conventional methods may remain viable in marine waters, and both plankton and marine sediments may serve as reservoirs for pathogenic organisms, which can emerge to become infective when conditions are favorable. In this paper we address the environmental factors that may contribute to illness, the types of associated economic costs, the issues of water quality monitoring and the policy implications raised by the apparent rise in incidence of marine water-related illnesses. PMID- 11485862 TI - An invertebrate model of the developmental neurotoxicity of insecticides: effects of chlorpyrifos and dieldrin in sea urchin embryos and larvae. AB - Chlorpyrifos targets mammalian brain development through a combination of effects directed at cholinergic receptors and intracellular signaling cascades that are involved in cell differentiation. We used sea urchin embryos as an invertebrate model system to explore the cellular mechanisms underlying the actions of chlorpyrifos and to delineate the critical period of developmental vulnerability. Sea urchin embryos and larvae were exposed to chlorpyrifos at different stages of development ranging from early cell cleavages through the prism stage. Although early cleavages were unaffected even at high chlorpyrifos concentrations, micromolar concentrations added at the mid-blastula stage evoked a prominent change in cell phenotype and overall larval structure, with appearance of pigmented cells followed by their accumulation in an extralarval cap that was extruded from the animal pole. At higher concentrations (20-40 microM), these abnormal cells constituted over 90% of the total cell number. Studies with cholinergic receptor blocking agents and protein kinase C inhibitors indicated two distinct types of effects, one mediated through stimulation of nicotinic cholinergic receptors and the other targeting intracellular signaling. The effects of chlorpyrifos were not mimicked by chlorpyrifos oxon, the active metabolite that inhibits cholinesterase, nor by nonorganophosphate cholinesterase inhibitors. Dieldrin, an organochlorine that targets GABA(A )receptors, was similarly ineffective. The effects of chlorpyrifos and its underlying cholinergic and signaling-related mechanisms parallel prior findings in mammalian embryonic central nervous system. Invertebrate test systems may thus provide both a screening procedure for potential neuroteratogenesis by organophosphate-related compounds, as well as a system with which to uncover novel mechanisms underlying developmental vulnerability. PMID- 11485863 TI - Human fatalities from cyanobacteria: chemical and biological evidence for cyanotoxins. AB - An outbreak of acute liver failure occurred at a dialysis center in Caruaru, Brazil (8 degrees 17' S, 35 degrees 58' W), 134 km from Recife, the state capital of Pernambuco. At the clinic, 116 (89%) of 131 patients experienced visual disturbances, nausea, and vomiting after routine hemodialysis treatment on 13-20 February 1996. Subsequently, 100 patients developed acute liver failure, and of these 76 died. As of December 1996, 52 of the deaths could be attributed to a common syndrome now called Caruaru syndrome. Examination of phytoplankton from the dialysis clinic's water source, analyses of the clinic's water treatment system, plus serum and liver tissue of clinic patients led to the identification of two groups of cyanobacterial toxins, the hepatotoxic cyclic peptide microcystins and the hepatotoxic alkaloid cylindrospermopsin. Comparison of victims' symptoms and pathology using animal studies of these two cyanotoxins leads us to conclude that the major contributing factor to death of the dialyses patients was intravenous exposure to microcystins, specifically microcystin-YR, LR, and -AR. From liver concentrations and exposure volumes, it was estimated that 19.5 microg/L microcystin was in the water used for dialysis treatments. This is 19.5 times the level set as a guideline for safe drinking water supplies by the World Health Organization. PMID- 11485864 TI - Endosulfan exposure disrupts pheromonal systems in the red-spotted newt: a mechanism for subtle effects of environmental chemicals. AB - Because chemicals introduced into the environment by humans can affect both long term survivorship and reproduction of amphibians, discovering the specific mechanisms through which these chemicals act may facilitate the development of plans for amphibian conservation. We investigated the amphibian pheromonal system as a potential target of common environmental chemicals. By treating female red spotted newts, Notophthalmus viridescens, to a commonly used insecticide, endosulfan, we found that the pheromonal system is highly susceptible to low concentration exposure. The impairment of the pheromonal system directly led to disrupted mate choice and lowered mating success. There were no other notable physiologic or behavioral changes demonstrated by the animals at the insecticide concentrations administered. Our findings suggest that the amphibian pheromonal system is one of the systems subject to subtle negative effects of environmental chemicals. PMID- 11485865 TI - Perinatal exposure to low doses of bisphenol A affects body weight, patterns of estrous cyclicity, and plasma LH levels. AB - The nonsteroidal estrogenic compound bisphenol A (BPA) is a monomer used in the manufacture of polycarbonate plastics and resins. BPA may be ingested by humans as it reportedly leaches from the lining of tin cans into foods, from dental sealants into saliva, and from polycarbonate bottles into their contents. Because BPA is weakly estrogenic--approximately 10,000-fold less potent than 17beta estradiol--current environmental exposure levels have been considered orders of magnitude below the dose required for adverse effects on health. Herein we demonstrate measurable effects on the offspring of Sprague-Dawley female rats that were exposed, via their drinking water, to approximately 0.1 mg BPA/kg body weight (bw)/day (low dose) or 1.2 mg BPA/kg bw/day (high dose) from day 6 of pregnancy through the period of lactation. Offspring exposed to BPA exhibited an increase in body weight that was apparent soon after birth and continued into adulthood. In addition, female offspring exposed perinatally to the high dose of BPA exhibited altered patterns of estrous cyclicity and decreased levels of plasma luteinizing hormone (LH) in adulthood. Administration of neither the doses of BPA that caused effects during perinatal exposure nor a 10-fold higher dose was able to evoke a uterotropic response in ovariectomized postpubertal females. These data indicate an increased sensitivity to BPA during the perinatal period and suggest the need for careful evaluation of the current levels of exposure to this compound. PMID- 11485866 TI - Experimental evaluation of vitellogenin as a predictive biomarker for reproductive disruption. AB - Vitellogenin (VTG) synthesis in male oviparous vertebrates is used as an indicator of environmental estrogen exposure, but the relationship between elevated VTG levels and the effects of environmental estrogens on reproductive success are poorly understood. To examine whether altered VTG expression predicts reproductive impairment, we exposed medaka (Oryzias latipes) for 2 or 8 weeks posthatch to 0, 0.5, 1.0, 2.5, and 7.5 ppb of the environmental estrogen o,p' DDT. Fish were sampled 2, 4, and 8 weeks after hatch to examine VTG expression and gonad development. After exposure, fish were transferred to clean water, grown to sexual maturity, and placed in mating pairs. We collected eggs for 7 days and scored them for fecundity (number of eggs), fertility (percent fertilized), and hatching success (percent hatched). DDT had no effect on VTG expression after a 2-week exposure, whereas all doses induced VTG after 8 weeks. At both exposure durations, the highest doses of DDT caused a female-skewed sex ratio in adults. Gonadal feminization appeared to be progressive: some ovotestes were observed after 2- or 4-week exposure to the two highest doses, but the proportion of ovaries increased after 8 weeks. Both 2- and 8-week exposures significantly reduced fertility and hatching success at all doses, with lower doses having a greater effect after longer exposure. Fertility and hatching success were more sensitive to estrogenic disruption than were gonad differentiation and vitellogenin expression. We suggest that VTG expression may be interpreted as a warning of reproductive consequences, but absence of expression cannot be interpreted as absence of consequences. PMID- 11485868 TI - Certain styrene oligomers have proliferative activity on MCF-7 human breast tumor cells and binding affinity for human estrogen receptor. AB - To examine the estrogenic activities of styrene oligomers, we carried out cell proliferation assays with estrogen-sensitive MCF-7 cells and competitive binding assays to human estrogen receptor [alpha] (hER[alpha]). The styrene oligomers tested were 1,3-diphenyl propane (SD-1), 2,4-diphenyl-1-butene (SD-2), cis-1,2 diphenyl cyclobutane (SD-3), trans-1,2-diphenyl cyclobutane (SD-4), 2,4,6 triphenyl-1-hexene (ST-1), 1a-phenyl-4a-(1'-phenylethyl)tetralin (ST-2), 1a phenyl-4e-(1'-phenylethyl)tetralin (ST-3), 1e-phenyl-4a-(1'-phenylethyl)tetralin (ST-4), 1e-phenyl-4e-(1'-phenylethyl)tetralin (ST-5), 1e,3e,5a triphenylcyclohexane (ST-6), and 1e,3e,5e-triphenylcyclohexane (ST-7). In the MCF 7 cell proliferation assay, styrene trimers (ST-1, ST-3, ST-4, and ST-5) had the highest proliferative activities of the compounds tested. The relative potency of these chemicals was 0.0002-0.0015%, which was comparable with that of bisphenol A (0.0001-0.0025%), and their relative proliferative effect was 51-104%. Styrene dimers (SD-3 and SD-4) also significantly increased the cell yields. However, SD 1, SD-2, ST-2, ST-6, and ST-7 had insignificant proliferative activities. The competitive binding assay revealed the binding affinity of some styrene oligomers for hER[alpha]. The order of their binding potency for hER[alpha] was as follows: ST-4 > ST-2 > ST-3 > ST-5 > ST-1 > SD-3 > SD-4 > SD-2 > SD-1. ST-6 and ST-7 did not appear to bind to hER[alpha]. The present studies indicate that styrene dimers SD-3 and SD-4 and styrene trimers ST-1, ST-3, ST-4, and ST-5 have estrogenic activity on MCF-7 cells and binding affinity for hER[alpha]. These compounds might be endocrine disrupters. PMID- 11485867 TI - Optimization of a yeast estrogen screen and its applicability to study the release of estrogenic isoflavones from a soygerm powder. AB - Here we describe a redesigned protocol of the yeast estrogen screen developed by Routledge and Sumpter. The redesigned test comprises two steps. First, a large amount of yeast with estrogenic compounds is incubated for 24 hr. Subsequently, a mixture of cycloheximide and the chromogenic substrate chlorophenol red-beta-d galactopyranoside (CPRG) is added. The cycloheximide stops protein synthesis and allows for an end-point measurement of beta-galactosidase activity generated during the first 24 hr. CPRG is converted to chlorophenol red and reflects beta galactosidase activity, which is indicative of the estrogenic activity. The modifications shorten the duration of the assay at least 1 day and avoid interference of the estrogenic CPRG or chlorophenol red. The redesigned and the original protocol were used to study the estrogenic activity of bisphenol A, methoxychlor, p,p'-DDT, and isoflavones (genistein, daidzein, and glycitein). Bisphenol A, methoxychlor, and genistein triggered higher levels of beta galactosidase activity in the redesigned protocol. Estrogenic activity of p,p' DDT could only be demonstrated with the redesigned protocol. Glycitein and daidzein failed to give a response with both protocols. We also studied deconjugation of beta-glycosidic isoflavones present in soygerm powder. Treatment of the soygerm powder with beta-glycosidase released isoflavones. The estrogenic response of the samples was confirmed with the redesigned protocol and correlated with the amount of genistein present. The release of isoflavones under conditions prevailing in the intestines was studied. Bacterial beta-glycosidase present in the large intestine released isoflavones, and moderate estrogenic activity could be demonstrated. PMID- 11485869 TI - The sex difference in tumor incidence is related to the female condition: models for Europe and Italy. AB - A remarkable aspect of cancer distribution in Europe is the large spatial variability of the male-female incidence ratio, from no difference up to 50%. Given the evidence of the predominantly environmental origin of cancer, we studied the ability of a set of socioeconomic indicators of the female condition to model the spatial variability of the sex difference in tumor incidence at two different scales: between countries (Europe) and between provinces (Italy). The sex difference in tumor incidence correlated with female socioeconomic condition indicators at the same extent (r = 0.73) in both situations, but in opposite directions. In the European study the higher the sexual social equality the lower the differential tumor incidence, whereas the opposite result was shown by the between-provinces Italian study. We also investigated the relation of the female condition indicator with other social and cultural descriptors of the same populations, and we suggest explanatory models linking female condition and pathology at the continental and local scales. Overall, our analysis supports the predominantly environmental origin of cancer and stresses the importance of relating cancer patterns to societal determinants. Our analysis also suggests that the sex difference in tumor incidence is a very useful probe for exploring the social-economic cultural correlates of cancer in human populations. We emphasize the need for a thorough analysis of the empirical correlations highlighted in ecologic studies. PMID- 11485870 TI - Acute exposure to environmental tobacco smoke and heart rate variability. AB - Environmental tobacco smoke (ETS) has been associated with cardiovascular mortality. Pathophysiologic pathways leading from ETS exposure to cardiopulmonary disease are still being explored. Reduced cardiac autonomic function, as measured by heart rate variability (HRV), has been associated with cardiac vulnerability and may represent an important pathophysiologic mechanism linking ETS and risk of cardiac mortality. In this study we evaluated acute ETS exposure in a commercial airport with changes in HRV in 16 adult nonsmokers. We conducted ambulatory electrocardiographic (ECG) monitoring for 8-hr periods while participants alternated 2 hr in nonsmoking and smoking areas. Nicotine and respirable suspended particle concentrations and participants' blood oxygen saturation were also monitored. We calculated time and frequency domain measures of HRV for periods in and out of the smoking area, and we evaluated associations with ETS using comparative statistics and regression modeling. ETS exposure was negatively associated with all measures of HRV. During exposure periods, we observed an average decrement of approximately 12% in the standard deviation of all normal-to normal heart beat intervals (an estimate of overall HRV). ETS exposures were not associated with mean heart rate or blood oxygen saturation. Altered cardiac autonomic function, assessed by decrements in HRV, is associated with acute exposure to ETS and may be part of the pathophysiologic mechanisms linking ETS exposure and increased cardiac vulnerability. PMID- 11485871 TI - Biomonitoring brevetoxin exposure in mammals using blood collection cards. AB - A method has been tested in laboratory mice to monitor for the presence of brevetoxins in blood after exposure. The use of blood collection cards is an adaptation of a method employed for routine diagnostic and genetic testing of newborns. Blood is collected and applied to a 0.5-inch diameter circle on a specially prepared blood collection card and allowed to dry. The blood spots are then extracted and the presence of toxin activity is first screened using a high throughput receptor binding assay. Positive samples are then examined for specific brevetoxin congeners by liquid chromatography-tandem mass spectrometry. Preliminary experiments tested the efficiency and linearity of toxin extraction from blood spiked with brevetoxin-3 (PbTx-3). Blood from treated mice was tested for the presence of brevetoxin at different times following exposure to a sublethal dose (180 microg/kg PbTx-3). Brevetoxin activity determined by receptor assay increased to 25 +/- 7.4 nM PbTx-3 equivalents within 4 hr after exposure and was still detectable in three of four animals 24 hr after exposure. Tandem mass spectrometry provided confirmation of PbTx-3, which also increased for the time points between 0.5 and 4.0 hr exposure. However, PbTx-3 was not detected at 24 hr, which suggested the formation of a biologically active metabolite. We anticipate that this approach will provide a method to biomonitor brevetoxins in living marine resources (e.g., finfish), protected species, and humans. PMID- 11485872 TI - Solid-tumor mortality in the vicinity of uranium cycle facilities and nuclear power plants in Spain. AB - To ascertain solid tumor mortality in towns near Spain's four nuclear power plants and four nuclear fuel facilities from 1975 to 1993, we conducted a mortality study based on 12,245 cancer deaths in 283 towns situated within a 30 km radius of the above installations. As nonexposed areas, we used 275 towns lying within a 50- to 100-km radius of each installation, matched by population size and sociodemographic characteristics (income level, proportion of active population engaged in farming, proportion of unemployed, percentage of illiteracy, and province). Using log-linear models, we examined relative risk for each area and trends in risk with increasing proximity to an installation. The results reveal a pattern of solid-tumor mortality in the vicinity of uranium cycle facilities, basically characterized by excess lung [relative risk (RR) 1.12, 95% confidence interval (CI), 1.02-1.25] and renal cancer mortality (RR 1.37, 95% CI, 1.07-1.76). Besides the effects of natural radiation, these results could well be evincing the influence on public health exerted by the environmental impact of mining. No such well-defined pattern appeared in the vicinity of nuclear power plants. Monitoring of cancer incidence and mortality is recommended in areas surrounding nuclear fuel facilities and nuclear power plants, and more specific studies are called for in areas adjacent to installations that have been fully operational for longer periods. In this regard, it is important to use dosimetric information in all future studies. PMID- 11485873 TI - Availability of polychlorinated biphenyls (PCBs) and lindane for uptake by intestinal Caco-2 cells. AB - Children may ingest contaminated soil from hand to mouth. To assess this exposure route, we need to know the oral bioavailability of the contaminants. Two determining steps in bioavailability of soil-borne contaminants are mobilization from soil during digestion, which is followed by intestinal absorption. The first step has been investigated in previous studies that showed that a substantial fraction of PCBs and lindane is mobilized from soil during artificial digestion. Furthermore, almost all contaminants are sorbed to constituents of artificial human small intestinal fluid (i.e., chyme), whereas only a small fraction is freely dissolved. In this study, we examine the second step using intestinal epithelial Caco-2 cells. The composition of the apical exposure medium was varied by addition of artificial chyme, bile, or oleic acid at similar or increasing total contaminant concentrations. The uptake curves were described by rate constants. The uptake flux seemed to be dose-dependent. Furthermore, different exposure media with similar total contaminant concentrations resulted in various uptake rates. This can be attributed to different freely dissolved concentrations and carrier effects. In addition, the large fractions of contaminants in the cells indicate that PCBs and lindane sorbed to bile, oleic acid, and digestive proteins contributed to the uptake flux toward the cells. These results can be extrapolated qualitatively to in vivo conditions. Because the sorbed contaminants should be considered available for absorption, the first step of mobilization from soil is the most important step for oral bioavailability of the presently investigated soil-borne contaminants. PMID- 11485875 TI - The secular trends in male:female ratio at birth in postwar industrialized countries. AB - Finnish investigators [Vartiainen et al. Environmental Chemicals and Changes in Sex Ratio: Analysis Over 250 Years in Finland. Environ Health Perspect 107:813 815 (1999)] presented the sex ratio of all newborn babies from 1751 to 1997 in order to evaluate whether Finnish long-term data are compatible with the hypothesis that the decrease in the ratio of male to female births after World War I and World War II in industrial countries is caused by environmental factors. They found an increase in the proportion of males from 1751 to 1920, which was interrupted by peaks in male births during World War I and World War II and followed by a decrease thereafter, similar to the trends in many other countries. The turning point of male proportion, however, preceded the period of industrialization and introduction of pesticides and hormonal drugs. Thus, a causal association between these environmental exposures and this decrease is unlikely. In addition, none of the various family parameters (e.g., paternal age, maternal age, age difference in parents, birth order) could explain the historical time trends. Vartiainen et al. concluded that at present it is unknown how these historical trends could be mediated. The postwar secular decline of the male:female ratio at birth is not an isolated phenomenon and parallels the decline of perinatal morbidity and mortality, congenital anomalies, and various constitutional diseases. This parallelism indicates a common etiology and may be caused by reduction of conceptopathology, as a correlate to increasing socioeconomic development. An inverted dose response or the dose-response fallacy due to vanishing male conceptuses explains the low sex ratios before World War I and World War II in newborns from black parents and from the lowest socioeconomic classes. PMID- 11485876 TI - Reduction of sperm motility in a male laboratory worker exposed to solvents: a case study. AB - A 34-year-old male laboratory worker suffered from asthenospermia and fertility problems. He was suspected of having been exposed to solvents used at work due to a malfunction of the ventilation system in his laboratory from August 1996 to April 1997. A laboratory walk-through and air and bulk sample collection were performed to determine the possible exposure levels of chemical hazards in his job. The scenario was reconstructed to simulate the worker's previous exposure during the ventilation shutdown period. It was found that the worker was possibly exposed to chloroform at levels of 10 or 50 times higher than the permissible exposure limit or the threshold limit value of 2 hr/day, 5.5 days/week, and 4.25 weeks/month for 8 months. Because chloroform is known to be spermatotoxic, the possibility of chloroform causing the worker's asthenospermia cannot be ruled out. Further study on spermatotoxicity of chloroform is warranted. PMID- 11485874 TI - Modulatory effects of neonatal exposure to TCDD, or a mixture of PCBs, p,p'-DDT, and p-p'-DDE, on methylnitrosourea-induced mammary tumor development in the rat. AB - The role of organochlorine (OC) exposure in the etiology of breast cancer remains controversial. Thus, our objective was to determine whether the most abundant and toxic OCs found in human milk could, when ingested during the neonatal period, modulate the development of mammary tumors in the rat. We prepared a mixture composed of p,p'-dichlorodiphenyltrichloroethane (DDT), its major metabolite, p,p'-dichlorodiphenyldichloroethene (DDE), and 19 polychlorinated biphenyls (PCB) based on their concentrations found in the milk of Canadian women. Neonate rats at 1, 5, 10, 15, and 20 days of age were gavaged with this mixture, at 10, 100, and 1,000 times the amount that a human baby would consume. An additional group received 2.5 microg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)/kg body weight (bw) by gavage at 18 days of age, instead of the mixture. On day 21, all treatment groups, except for a control group and a 1,000-mix group, received a single intraperitoneal injection of methylnitrosourea (MNU, 30 mg/kg bw), the initiator of the carcinogenic process. The average number of rats per treatment group was 33. Rats were sacrificed when their tumors reached 1 cm in size, or at 308 days of age. We prepared mammary tumors and mammary gland whole mounts for histologic analysis. There were no significant effects when only the malignant or only the benign tumors were considered. After all benign and malignant lesions were pooled, the number of mammary tumors differed among all MNU-treated groups (p = 0.02) with more lesions developing in the MNU-1,000[times] (median = 4.5; p = 0.05) and MNU-TCDD (median = 5.5; p = 0.07) compared to the MNU-0 rats (median = 2). Compared to the MNU-0 group, the percentage of rats that developed palpable tumors (benign plus malignant) was slightly higher (p = 0.06) in the MNU-TCDD group, but not in the MNU-1,000[times] group. The percentage of palpable tumors that were malignant was higher (p = 0.02) in the MNU-100[times] group (15/16, 94%) than in the MNU-0 group (10/18, 56%). The highest dose of the mixture delayed (p = 0.03) the development of tumors, but this was not observed with the MNU-TCDD treatment. These results suggest that neonatal exposure to high doses of organochlorines could favor the development of MNU-induced mammary lesions, but also delays the development of palpable tumors in the rat. PMID- 11485877 TI - Environmental health in space. PMID- 11485878 TI - DDT risk assessments. PMID- 11485879 TI - Bringing new life to the dead zone. PMID- 11485880 TI - HACCP hassles. PMID- 11485881 TI - African trade agreement--not entirely free? PMID- 11485882 TI - Human DNA polymerase iota: wrong for the right reasons. PMID- 11485883 TI - New research group to target autoimmune disorders. PMID- 11485884 TI - Livestock farming: eating up the environment? PMID- 11485885 TI - Aquaculture: satisfying the global appetite. PMID- 11485886 TI - Serving up food safety: who wants a piece of the pie? AB - A total of 12 federal agencies, plus their state counterparts, contribute to the regulatory snarl that governs the safety of the American food supply. With so much federal oversight, one might expect U.S. foods to be virtually risk-free. But this is hardly the case; contaminated food is responsible for 75 million illnesses, 325,000 hospitalizations, and 5,000 deaths in the United States each year. Recent reports from the General Accounting Office and the National Research Council claim that creation of a single agency with centralized authority is the best solution to U.S. food safety problems. Some experts agree that regulatory gaps in food safety highlight the need for centralized leadership, and that more money is necessary to fund the number of inspectors needed to adequately inspect the food supply before it reaches consumers. The single-agency concept has garnered congressional, industry, and scientific support, but the idea isn't without its skeptics, who believe that consolidating food safety under a single agency eliminates checks and balances offered by the current system and, more importantly, runs the risk of politicizing the agency. PMID- 11485887 TI - From pigsties to hog heaven? AB - In the continuing transformation of U.S. agriculture, North Carolina finds itself on the front edge of change. Between 1989 and 1998, the number of hogs in the state's pork industry quintupled---and so has the amount of hog waste that must be disposed of. Now the state has engaged private and public resources in a rapid search for better ways for handling hog waste. A technology review panel has approved the first round of proposals for a number of novel technologies to be developed through funds from a government-industry agreement. A second batch of proposals is expected to be approved by late summer. PMID- 11485889 TI - Oral health status of kindergarten children in a medium-sized German town near the Czech border. AB - A cohort examination was performed to investigate the oral health status of 215 kindergarten children in a medium-sized German town near the Czech border. Despite an extensive campaign involving information events and discussions, only 130 of these children could be recruited to participate in the study. They showed a mean caries rate of 50%, which was up to 60% among the 6-year-olds. These findings clearly fall short of the WHO goals for 2000 and indicate that the geographic area covered by the study has a significant oral health deficit. The study also showed a particularly high prevalence of carious defects in a small number of children. The posterior occlusal relations were normal in 90.8 percent of cases. Deformations of the os maxillare (open bite due to finger sucking) were found in 20% of the 4 year group, in 2.3% of the 5 year group, and in 13% of the 6 year group. PMID- 11485888 TI - Antibiotic therapy in intra-abdominal infections--a review on randomised clinical trials. AB - There have been 79 randomised antibiotic studies in intra-abdominal infections retrieved. The overall success rate of the studied antibiotics ranges from 70 100%. Unfortunately only about one fourth of the studies have used a disease severity classification, e.g., APACHE II score, despite clear recommendations by the Surgical Infections Society of North America. The mortality rate in the published antibiotic studies is still rather low (approximately 4%) and does not correspond to the average mortality in peritonitis (30-40%). Failure analysis is not uniform and only performed in about 1/5 of retrieved studies. Failure analysis included data on diagnosis, type of operation, pathogen isolated at first operation, susceptibility and persistence of pathogen, re-operation or change of antibiotic regimen, and follow-up (ICU duration, death or survival, hospitalisation). Only one study has performed an analysis of the adequacy of the surgical treatment (source control). The clinical success rate of the antibiotics studied in a larger population is comparable for gentamicin + clindamycin (80%), tobramycin + clindamycin (83%), meropenem (89%), imipenem (85%), aztreonam + clindamycin (89%), cefoxitin (88%), cefotetan (92%), moxalactam (83%), cefotaxime + metronidazole (87%), ampicillin/sulbactam (87%). Piperacillin/tazobactam has in most studies a success rate of approximately 90%. The aggregated data on adverse events and clinical failure rate do not show a major advantage for any of these antibiotics. It is striking that the adverse event rate reported for ticarcillin/clavulanic acid is low when compared to all other antibiotics, which is in contrast to severe adverse events reported for clavulanic acid. The data of quinolone studies in intra-abdominal infections do not yet allow a recommendation, even when it is acknowledged that two studies were performed with good results and a good study plan. In conclusion, the comparability of antibiotic studies in intra-abdominal infections is limited due to a lack of disease severity stratification and a relatively small study population for most antibiotics. The clinical success rate of the best-studied antibiotics is similar and the choice which antibiotic is used depends on the expected pathogens and the resistance rate in a clinical setting. PMID- 11485890 TI - New surface biopolymers for oxygenators: an in vitro hemocompatibility test of poly(2-methoxyethylacrylate). AB - BACKGROUND/OBJECTIVE: The continuous interaction of blood with artificial contact surfaces under cardiopulmonary bypass can lead to a substantial damage of blood cells and plasma factors. Surface biopolymers in oxygenation systems can help increasing the hemocompatibility, often combined with anticoagulative agents such as heparin. The poly(2-methoxyethylacrylate) [PMEA] is a new heparin-free polymer. The objective of this experimental study was to evaluate the hemocompatibility of a PMEA-coated oxygenator (Terumo Capiox RX-25) (PTX) under standardized in vitro conditions compared to two ionic-bound and one covalent bound heparin-coated models. METHOD: Each oxygenator was mounted in a separate standardized closed circulation system. Heparinized (5 IE/mL) fresh human blood from the same donor (hemodilution: Ringer's solution) was used. Circulation time: 120 - 180 min with a flow rate of 4.0 L/min. Blood samplings: at the beginning, 5. min and every 30 min of the circulation. PARAMETERS: platelets, granulocytes, plasma factors (p-selectin, alpha-granulomeres expression, and TAT(III)-complex). After the experiment, oxygenators were dismantled and examined by scanning electron microscopy. RESULTS: All of the oxygenators led to an initial reduction of platelets and granulocytes. PTX had the lowest platelet and granulocyte reduction rates. With a lower p-selectin release compared to covalent-bound heparin-coated oxygenator and higher expression of alpha-granulomeres compared to ionic-bound heparin-coated oxygenators, the results of PTX indicated that a high number of circulating platelets were intact on the PMEA surface. TAT(III)-complex showed a steady increase in all of the oxygenators during the tests, more remarkably in PTX. In contrast to ionic-bound heparin-coated oxygenators, the electron microscopy displayed virtually no cellular accumulation on hollow fiber and housing surfaces of PTX and covalent-bound heparin-coated oxygenator. CONCLUSIONS: 1. The hemocompatibility characteristics of PTX were remarkably better than ionic-bound heparin-coated oxygenators and slightly better than the covalent-bound heparin-coated model under in vitro conditions. 2. The PMEA coating can be a useful alternative for patients with heparin-associated disorders. 3. The clinical feasibility of PTX should be evaluated under in vivo conditions. PMID- 11485891 TI - Therapeutic and prophylactic effects of crude honey on chronic seborrheic dermatitis and dandruff. AB - Honey has antibacterial, antifungal and antioxidants activities and has high nutrient value. In this study we investigated the potential use of topical application of crude honey in the management of seborrheic dermatitis and dandruff. Thirty patients with chronic seborrheic dermatitis of scalp, face and front of chest were entered for study. Twenty patients were males and 10 were females, their ages ranged between 15 and 60 years. The patients had scaling, itching and hair loss. The lesions were scaling macules, papules and dry white plaques with crust and fissures. The patients were asked to apply diluted crude honey (90% honey diluted in warm water) every other day on the lesions with gentle rubbing for 2-3 mins. Honey was left for 3 hr before gentle rinsing with warm water. The patients were followed daily for itching, scaling, hair loss and the lesions were examined. Treatment was continued for 4 weeks. The improved patients were included in a prophylactic phase, lasting six months. Half patients were treated with the topical honey once weekly and the other half served as control. All the patients responded markedly with application of honey. Itching was relieved and scaling was disappeared within one week. Skin lesions were healed and disappeared completely within 2 weeks. In addition, patients showed subjective improvement in hair loss. None of the patients ( 15 patients) treated with honey application once weekly for six months showed relapse while the 12/15 patients who had no prophylactic treatment with honey experienced a relapse of the lesions 2-4 months after stopping treatment. It might be concluded that crude honey could markedly improve seborrheic dermatitis and associated hair loss and prevent relapse when applied weekly. PMID- 11485892 TI - Plastic surgical management in tissue extravasation of cytotoxic agents in the upper extremity. AB - Extravasation injuries in subcutaneous tissues during peripheral intravenous administration can result in extensive soft-tissue defects. Early treatment (surgical removal of the extravasated material within 24 h) and late treatment (debridement and coverage) are two possible approaches for this kind of injury. Eighteen patients who suffered a significant extravasation injury were treated surgically between 1/1996 and 5/2001. All patients were referred late (mean 22 days after the event) to our clinic with soft-tissue defects or skin necrosis and were accordingly only amenable to late treatment. The patients with defects over the dorsum of the hand, forearm or cubital fossa area underwent debridement, temporary wound coverage and skin grafting or coverage with a local flap. Spotted infiltrating lesions and necrosis are typical histologic features of a cytotoxic agent extravasation. A safe margin of resection is only achievable with a wide three dimensional excision. The healing time of defects was a mean of 15 days after either skin grafting or flap coverage with no significant difference between the two differing treatment groups. Early referral of patients with extravasation injuries to a specialized department for plastic and hand surgery may in future enable earlier surgical treatment, reduce time of illness and costs. PMID- 11485893 TI - S680N substitution of the follicle-stimulating hormone receptor is a common polymorphism not associated with spontaneous human twinning. AB - OBJECTIVE: To determine the influence of the A307/S680 and T307/N680 isoforms of the follicle-stimulating hormone receptor (FSHR) gene on the incidence of spontaneous human twinning. DESIGN: Case-control study. SETTING: Departments of Obstetrics and Gynecology and of Clinical Chemistry-Grosshadern, University Hospital Munich, Germany. PATIENT(S): Fifty-four mothers with dichorionic twin pregnancies and 92 singleton mothers as controls, who had conceived without assisted reproduction. INTERVENTION(S): Exon 10 of the FSHR gene was screened for the G2105A/S680N mutation. MAIN OUTCOME MEASURE(S): Amplification of genomic DNA by the polymerase chain reaction followed by restriction fragment length polymorphism analysis. RESULT(S): Allele frequencies for the G2105A/ S680N substitution of the FSHR in twin mothers were not different from those of controls (genotype (isoform) [twins vs. controls]: G/G (S/S) [24.1% vs. 22.3%]; A/G (N/S) [57.4% vs. 55.4%]; A/A (N/N) [18.5% vs. 22.3%]). Subgroup analysis of women with three or more successful pregnancies gave a similar result (G/G (S/S) [17.7% vs. 13.7%]; A/G (N/S) [64.7% vs. 63.6%]; A/A (N/N) [17.6% vs. 22.7%]). There was no correlation between FSHR isoform and twinning. CONCLUSION(S): The S680N substitution of the follicle-stimulating hormone receptor is a common polymorphism not associated with spontaneous human twinning. PMID- 11485895 TI - New insights into the role of nuclear factor-kappaB in cell growth regulation. AB - The nuclear factor (NF)-kappaB family of eukaryotic transcription factors plays an important role in the regulation of immune response, embryo and cell lineage development, cell apoptosis, cell-cycle progression, inflammation, and oncogenesis. A wide range of stimuli, including cytokines, mitogens, environmental particles, toxic metals, and viral or bacterial products, activate NF-kappaB, mostly through IkappaB kinase (IKK)-dependent phosphorylation and subsequent degradation of its inhibitor, the IkappaB family of proteins. Activated NF-kappaB translocates into the nucleus where it modulates the expression of a variety of genes, including those encoding cytokines, growth factors, acute phase response proteins, cell adhesion molecules, other transcription factors, and several cell apoptosis regulators. During the past few years, tremendous progress has been achieved in our understanding on how intracellular signaling pathways are transmitted in either a linear or a network manner leading to the activation of NF-kappaB and subsequent cell growth control. However, a detailed molecular mechanism of NF-kappaB regulating cell growth has yet to be determined. Elucidation of the relationships between NF-kappaB activation and cell growth will be important in developing new strategies for the treatment of various human diseases, such as chronic autoimmune disorder and cancer. PMID- 11485896 TI - Inflammation in nonhealing diabetic wounds: the space-time continuum does matter. PMID- 11485897 TI - Complex regulation of telomerase activity: implications for cancer therapy. PMID- 11485898 TI - Fusion of the ALK gene to the clathrin heavy chain gene, CLTC, in inflammatory myofibroblastic tumor. AB - Inflammatory myofibroblastic tumor (IMT) is a rare, but distinctive mesenchymal neoplasm composed of fascicles of bland myofibroblasts admixed with a prominent inflammatory component. Genetic studies of IMTs have demonstrated chromosomal abnormalities of 2p23 and rearrangement of the anaplastic lymphoma kinase (ALK) gene locus. In a subset of IMTs, the ALK C-terminal kinase domain is fused with a tropomyosin N-terminal coiled-coil domain. In the current study, fusion of ALK with the clathrin heavy chain (CTLC) gene localized to 17q23 was detected in two cases of IMT. One of these cases exhibited a 2;17 translocation in addition to other karyotypic anomalies [46,XX,t(2;17)(p23;q23),add(16)(q24)]. PMID- 11485899 TI - Expression of interleukin-15 in mouse and human atherosclerotic lesions. AB - Atherosclerotic lesions are characterized by prominent macrophage and T-cell infiltration and atherosclerosis is widely recognized as an inflammatory disease. The cytokine interleukin-15 (IL-15) has T-cell chemotactic and pro-inflammatory properties and promotes the recruitment of T cells to sites of inflammation. We have therefore examined IL-15 expression in the atherosclerotic ApoE-deficient mouse model as well as in human atherosclerotic lesions. In gene expression arrays, a transcript corresponding to IL-15 mRNA was elevated in atherosclerotic aortas of ApoE-deficient mice fed a Western diet for 10 and 20 weeks, corresponding to lesions of the fatty streak and fibrofatty plaque stage, respectively. Immunostaining for IL-15 localized to aortic smooth muscle cells in nonatherosclerotic C57BL/6 mice, whereas both macrophages and smooth muscle cells stained positive for IL-15 in atherosclerotic lesions of ApoE-deficient mice. Finally, advanced atherosclerotic lesions of human carotid arteries were immunostained to determine whether IL-15 is involved in human disease. IL-15 protein was present also in the human lesions with a distribution primarily overlapping that of macrophages. In conclusion, IL-15 is up-regulated in both human and animal atherosclerotic lesions and may contribute to the recruitment of T cells and their activation during atherogenesis. PMID- 11485900 TI - Detection of cyclin D1 overexpression by real-time reverse-transcriptase-mediated quantitative polymerase chain reaction for the diagnosis of mantle cell lymphoma. AB - The diagnosis of mantle cell lymphoma (MCL) is particularly important for clinical management because of a remarkable prognostic difference between MCL and other types of B-cell lymphoma. In addition to immunohistochemical analysis, we have established a 5' exonuclease-based real-time reverse transcriptase-mediated quantitative polymerase chain reaction (RQ-PCR) method to detect cyclin D1 overexpression for the diagnosis of MCL. The RQ-PCR could detect cyclin D1 overexpression in all nine examined MCL cases, in contrast genomic PCR detected t(11;14) in only two of nine cases. By RQ-PCR the expression of G6PDH was significantly higher in myeloid leukemias than those in B-cell lymphomas (P = 0.018). As a result, cyclin D1/G6PDH ratio ranged from 0.78 to 12.4 (mean, 1.83) in MCL, exclusively higher than those in other B-cell lymphoma (0.00009 approximately 0.16) and myeloid leukemia (0.00011 approximately 0.085). The high expression of cyclin D1 in certain myeloid leukemias was identified to reflect their proliferative activity and not to represent the oncogenic overexpression. The 95% confidence interval of the cyclin D1/G6PDH ratio was 0.29 approximately 11.1 for MCL, 0.014 approximately 0.25 for other B-cell lymphomas and 0.000014 approximately 0.083 for myeloid leukemia, suggesting that a cutoff value can be set at 0.25. The RQ-PCR of cyclin D1 is convenient and especially useful for the diagnosis of MCL. PMID- 11485901 TI - AKT1/PKBalpha kinase is frequently elevated in human cancers and its constitutive activation is required for oncogenic transformation in NIH3T3 cells. AB - Extensive studies have demonstrated that the Akt/AKT1 pathway is essential for cell survival and inhibition of apoptosis; however, alterations of Akt/AKT1 in human primary tumors have not been well documented. In this report, significantly increased AKT1 kinase activity was detected in primary carcinomas of prostate (16 of 30), breast (19 of 50), and ovary (11 of 28). The results were confirmed by Western blot and immunohistochemical staining analyses with phospho-Ser473 Akt antibody. The majority of AKT1-activated tumors are high grade and stage III/lV (13 of 16 prostate, 15 of 19 breast, and 8 of 11 ovarian carcinomas). Previous studies showed that wild-type AKT1 was unable to transform NIH3T3 cells. To demonstrate the biological significance of AKT1 activation in human cancer, constitutively activated AKT1 (Myr-Akt) was introduced into NIH3T3 cells. Overexpression of Myr-Akt in the stably transfected cells resulted in malignant phenotype, as determined by growth in soft agar and tumor formation in nude mice. These data indicate that AKT1 kinase, which is frequently activated in human cancer, is a determinant in oncogenesis and a potential target for cancer intervention. PMID- 11485902 TI - Immunization with a nontoxic/nonfibrillar amyloid-beta homologous peptide reduces Alzheimer's disease-associated pathology in transgenic mice. AB - Transgenic mice with brain amyloid-beta (Abeta) plaques immunized with aggregated Abeta1-42 have reduced cerebral amyloid burden. However, the use of Abeta1-42 in humans may not be appropriate because it crosses the blood brain barrier, forms toxic fibrils, and can seed fibril formation. We report that immunization in transgenic APP mice (Tg2576) for 7 months with a soluble nonamyloidogenic, nontoxic Abeta homologous peptide reduced cortical and hippocampal brain amyloid burden by 89% (P = 0.0002) and 81% (P = 0.0001), respectively. Concurrently, brain levels of soluble Abeta1-42 were reduced by 57% (P = 0.0019). Ramified microglia expressing interleukin-1beta associated with the Abeta plaques were absent in the immunized mice indicating reduced inflammation in these animals. These promising findings suggest that immunization with nonamyloidogenic Abeta derivatives represents a potentially safer therapeutic approach to reduce amyloid burden in Alzheimer's disease, instead of using toxic Abeta fibrils. PMID- 11485903 TI - Expression of lysosome-associated membrane proteins in human colorectal neoplasms and inflammatory diseases. AB - The lysosome-associated membrane proteins (LAMPs)-1 and -2 are major constituents of the lysosomal membrane. These molecules are known to be among the most glycosylated proteins of several types of cells and cancer cells, and their expression in cancer cells is marked by a distinct difference in the structures of the oligosaccharides as compared to nonmalignant cells. We analyzed by immunohistochemistry the intensity and distribution of LAMP-1 and LAMP-2 in 9 human colorectal cancer cases and in 16 control cases, including inflammatory diseases (diverticulitis, ulcerative colitis, and Crohn's disease). LAMP proteins were expressed more intensely in the epithelium of colorectal neoplasms than in normal mucosa (P < 0.05), and no significant differences were found between adenoma and cancer cells (P > 0.05) in the same tissue section. Further, in sites of inactive inflammatory diseases and nonneoplastic areas in cancer specimens, no significant increases in epithelial LAMP proteins were observed, even in the proliferative zone of the lower crypt epithelium. Northern blot analysis showed increased expression of LAMP-1 and LAMP-2A in two of three colorectal cancers examined and increased LAMP-2B in all three cancers. Our findings suggest that LAMPs are related to neoplastic progression, but there is no direct association between the expression of LAMP molecules and cell proliferation. PMID- 11485905 TI - Hypomethylation of chromosome 1 heterochromatin DNA correlates with q-arm copy gain in human hepatocellular carcinoma. AB - Using comparative genomic hybridization (CGH) analysis, we, and others, have shown that there is a high and consistent incidence of chromosome 1q copy gain in human hepatocellular carcinoma (HCC). Chromosome 1 rearrangements, that involved peri-centromeric breakpoints, have also been frequently reported in karyotypic studies of HCC. Satellite DNA hypomethylation has been postulated as the mechanism underlying the induction of chromosome 1 peri-centromeric instability in many human cancers and in individuals with the rare recessive disorder ICF (immunodeficiency, centromeric heterochromatin instability, facial anomalies). In this study, we have investigated the role of DNA hypomethylation in 1q copy gain in HCC by examining the methylation status of chromosome 1 heterochromatin DNA (band 1q12). Thirty-six histologically confirmed samples of HCC were studied (24 paired tumor and adjacent nontumorous liver tissues, and 12 tumor only). Hypomethylation of satellite 2 (Sat2) DNA in 1q12 was analyzed by Southern blotting using methyl-sensitive enzyme digestion. In parallel, all cases were analyzed by CGH. A strong correlation between hypomethylated Sat2 sequences and 1q copy gain with a 1q12 breakpoint was found (P < 0.001). We postulate that such hypomethylation alters the interaction between the CpG-rich satellite DNA and chromatin proteins, resulting in heterochromatin decondensation, breakage and aberrant 1q formation. Spectral karyotyping further supported the presence of fragile 1q12 in HCC. Of particular interest was the finding of Sat2 DNA hypomethylation in 5 of 24 adjacent nontumorous liver tissues examined. These tissues showed no evidence of malignancy on histological examination nor did they display any CGH abnormalities. Our findings suggest a role for Sat2 demethylation in the early stages of the stepwise progression of liver carcinogenesis. PMID- 11485904 TI - Wound healing in MIP-1alpha(-/-) and MCP-1(-/-) mice. AB - A salient feature of normal wound healing is the development and resolution of an acute inflammatory response. Although much is known about the function of inflammatory cells within wounds, little is known about the chemotactic and activation signals that influence this response. As the CC chemokines macrophage inflammatory protein-1alpha (MIP-1alpha) and monocyte chemotactic protein-1 (MCP 1) are abundant in acute wounds, wound repair was examined in MIP-1alpha(-/-) and MCP-1(-/-) mice. Surprisingly, wound re-epithelialization, angiogenesis, and collagen synthesis in MIP-1alpha(-/-) mice was nearly identical to wild-type controls. In contrast, MCP-1(-/-) mice displayed significantly delayed wound re epithelialization, with the greatest delay at day 3 after injury (28 +/- 5% versus 79 +/- 14% re-epithelialization, P < 0.005). Wound angiogenesis was also delayed in MCP-1(-/-) mice, with a 48% reduction in capillary density at day 5 after injury. Collagen synthesis was impeded as well, with the wounds of MCP-1(-/ ) mice containing significantly less hydroxyproline than those of control mice (25 +/- 3 versus 50 +/- 8 microg/wound at day 5, P < 0.0001). No change in the number of wound macrophages was observed in MCP-1(-/-) mice, suggesting that monocyte recruitment into wounds is independent of this chemokine. The data suggest that MCP-1 plays a critical role in healing wounds, most likely by influencing the effector state of macrophages and other cell types. PMID- 11485906 TI - A novel method for isolation of neutrophils from murine blood using negative immunomagnetic separation. AB - Inappropriate neutrophil activation has been implicated in the pathology of several clinically important inflammatory conditions. Although murine models are extensively used in the investigation of such pathological processes, a reliable method by which viable, quiescent neutrophils can be isolated from murine blood has not been developed. Here we describe a novel method based on negative immunomagnetic separation, which yields highly pure populations of murine neutrophils. Blood is incubated with a cocktail of antibodies against specific cell markers on unwanted cells, and then with secondary antibody-coated magnetic beads. After running the preparation through a column within a magnetic field, labeled cells are retained, and a neutrophil-rich effluent is collected. This method yields a >95% pure suspension of >97% viable neutrophils, recovering approximately 70% of neutrophils from whole blood. Flow cytometric analysis shows little difference in surface L-selectin and CD18 expression on isolated neutrophils compared with neutrophils in whole blood, indicating that neutrophils are minimally activated bythe isolation process. Stimulation with phorbol 12 myristate 13-acetate (PMA) reduced L-selectin andincreased CD18 expression. Isolated neutrophilsmigrate under agarose in response to fMLP, and fluorescently labeled neutrophils transfused into recipient mice interact with postcapillary venules in a manner comparable to endogenous leukocytes. These findings show that neutrophils isolated using this method can be used for inflammatory studies in vitro and in vivo. PMID- 11485907 TI - The generation and characterization of a cell line derived from a sporadic renal angiomyolipoma: use of telomerase to obtain stable populations of cells from benign neoplasms. AB - Angiomyolipomas are benign tumors of the kidney derived from putative perivascular epithelioid cells, that may undergo differentiation into cells with features of melanocytes, smooth muscle, and fat. To gain further insight into angiomyolipomas, we have generated the first human angiomyolipoma cell line by sequential introduction of SV40 large T antigen and human telomerase into human angiomyolipoma cells. These cells show phenotypic characteristics of angiomyolipomas, namely differentiation markers of smooth muscle (smooth muscle actin), adipose tissue (peroxisome proliferator-activator receptor gamma, PPARgamma), and melanocytes (microophthalmia, MITF), thus demonstrating that a single cell type can exhibit all of these phenotypes. These cells should serve as a valuable tool to elucidate signal transduction pathways underlying renal angiomyolipomas. PMID- 11485908 TI - Ganglioside GM2/GD2 synthetase mRNA is a marker for detection of infrequent neuroblastoma cells in bone marrow. AB - GalNAcbeta1-4(NeuAcalpha2-3)Galbeta1-4Glcbeta1-Cer (GM2)/GalNAcbeta1 4(NeuAcalpha2-8NeuAcalpha2-3)Galbeta1-4Glcbeta1-1Cer (GD2) synthetase [beta-1,4-N acetyl-galactosaminyl transferase (GalNAc-T)] mRNA, which encodes a key glycosyltransferase for ganglioside GD2 synthesis, was assessed as a molecular marker for detecting metastatic neuroblastoma cells in bone marrow (BM). GalNAc-T mRNA expression by neuroblastoma cell lines (n = 15), primary untreated neuroblastoma tumors (n = 29), morphologically normal BM (n = 22), peripheral blood stem cells (n = 10) from patients with cancers other than neuroblastoma, and blood mononuclear cells from normal donors (n = 17) was assessed by using reverse transcriptase-polymerase chain reaction (RT-PCR) and electrochemiluminescence detection assay (RT-PCR/ECL). BM harvested from 15 neuroblastoma patients was tested before and after ex vivo immunomagnetic bead purging, and results were compared to immunocytological analysis of the same specimens. All neuroblastoma cell lines (mean, 653 x 10(3) ECL units) and primary tumors (mean, 683 x 10(3) ECL units) were positive for significant expression of GalNAc-T mRNA compared to normal blood and BM cells. The RT-PCR/ECL assay could detect GalNAc-T mRNA in 100 pg of total RNA, and in a mixture of one neuroblastoma cell among 10(7) normal BM or blood cells. Eight of 15 autologous BM cells harvested from patients with neuroblastoma had tumor cells detectable by immunocytology, and all 15 were positive for GalNAc-T mRNA. After ex vivo purging, none of the BM cells was immunocytology-positive, but six remained positive by the RT-PCR/ECL assay. GalNAc-T mRNA provides a specific and sensitive molecular marker for RT-PCR/ECL detection of infrequent neuroblastoma cells in BM. PMID- 11485909 TI - The perifollicular and marginal zones of the human splenic white pulp : do fibroblasts guide lymphocyte immigration? AB - We investigate the white pulp compartments of 73 human spleens and demonstrate that there are several microanatomical peculiarities in man that do not occur in rats or mice. Humans lack a marginal sinus separating the marginal zone (MZ) from the follicles or the follicular mantle zone. The MZ is divided into an inner and an outer compartment by a special type of fibroblasts. An additional compartment, termed the perifollicular zone, is present between the follicular MZ and the red pulp. The perifollicular zone contains sheathed capillaries and blood-filled spaces without endothelial lining. In the perifollicular zone, in the outer MZ, and in the T cell zone fibroblasts of an unusual phenotype occur. These cells stain for the adhesion molecules MAdCAM-1, VCAM-1 (CD106), and VAP-1; the Thy-1 (CD90) molecule; smooth muscle alpha-actin and smooth muscle myosin; cytokeratin 18; and thrombomodulin (CD141). They are, however, negative for the peripheral node addressin, the cutaneous lymphocyte antigen, CD34, PECAM-1 (CD31), and P- and E-selectin (CD62P and CD62E). In the MZ the fibroblasts are often tightly associated with CD4-positive T lymphocytes, whereas CD8-positive cells are almost absent. Our findings lead to the hypothesis, that recirculating CD4-positive T lymphocytes enter the human splenic white pulp from the open circulation of the perifollicular zone without crossing an endothelium. Specialized fibroblasts may attract these T cells and guide them into the periarteriolar T cell area. PMID- 11485910 TI - Blockade of receptor for advanced glycation end-products restores effective wound healing in diabetic mice. AB - Receptor for advanced glycation end-products (RAGE), and two of its ligands, AGE and EN-RAGEs (members of the S100/calgranulin family of pro-inflammatory cytokines), display enhanced expression in slowly resolving full-thickness excisional wounds developed in genetically diabetic db+/db+ mice. We tested the concept that blockade of RAGE, using soluble(s) RAGE, the extracellular ligand binding domain of the receptor, would enhance wound closure in these animals. Administration of sRAGE accelerated the development of appropriately limited inflammatory cell infiltration and activation in wound foci. In parallel with accelerated wound closure at later times, blockade of RAGE suppressed levels of cytokines; tumor necrosis factor-alpha; interleukin-6; and matrix metalloproteinases-2, -3, and -9. In addition, generation of thick, well vascularized granulation tissue was enhanced, in parallel with increased levels of platelet-derived growth factor-B and vascular endothelial growth factor. These findings identify a central role for RAGE in disordered wound healing associated with diabetes, and suggest that blockade of this receptor might represent a targeted strategy to restore effective wound repair in this disorder. PMID- 11485911 TI - Differential expression of BCL-2 family proteins in ALK-positive and ALK-negative anaplastic large cell lymphoma of T/null-cell lineage. AB - Anaplastic large-cell lymphoma (ALCL) of T- or null-cell lineage, as defined in the revised European-American lymphoma classification, includes a subset of tumors that carry the t(2;5)(p23;q35) resulting in overexpression of anaplastic lymphoma kinase (ALK). Patients with ALK+ ALCL are reported to have a better prognosis than patients with ALK- ALCL. Because the mechanisms for this survival difference are unknown, we investigated the hypothesis that apoptotic pathways may be involved. We therefore assessed expression levels of the anti-apoptotic proteins BCL-2 and BCL-XL and the pro-apoptotic proteins BAX and BCL-XS in T/null cell ALCL using immunohistochemical methods and correlated the findings with ALK expression and apoptotic rate (AR), the latter assessed by a modified Tdt mediated dUTP nick-end labeling assay. ALK was detected in 21 of 66 (31.8%) ALCLs. BCL-2 was not detected in 21 ALK+ ALCLs but was present in 26 of 45 (57.8%) ALK- ALCLs (P < 0.0001). ALK+ and ALK- ALCLs also showed significant differences in expression of BCL-XL, BAX, and BCL-XS. ALK+ tumors less commonly had a high level of BCL-XL (1 of 17 versus 14 of 35, P = 0.01), and more commonly had high levels of BAX (13 of 18 versus 15 of 36, P = 0.05), and BCL-XS (11 of 16 versus 12 of 31, P = 0.05) compared with ALK- tumors. ALK+ tumors also had a higher mean AR than ALK- tumors (3.4% versus 1.1%, P = 0.0002). Differential expression of BCL-2 family proteins may be responsible for the higher AR observed in ALK+ ALCL and provides a possible biological explanation for the better prognosis reported for patients with ALK+ ALCL. PMID- 11485912 TI - cis Interaction of the cell adhesion molecule CEACAM1 with integrin beta(3). AB - CEACAM1 is a cell adhesion molecule that has been implicated in a number of physiological processes (eg, tumor suppressor in epithelial tissues, potent angiogenic factor in microvessel formation, microbial receptor in human granulocytes and epithelial cells). The mechanism of CEACAM1 action is still largely unresolved but recent findings demonstrated that the cytoplasmic CEACAM1 domain is linked indirectly to the actin-based cytoskeleton. We have isolated integrin beta(3) as an associated protein using CEACAM1 tail affinity purification. This association depends on phosphorylation of Tyr-488 in the CEACAM1 cytoplasmic domain. Confocal laser scanning microscopy confirmed in vivo colocalization of both molecules in human granulocytes and epithelial cells. Furthermore, the concentrated colocalization at the tumor-stroma interface of invading melanoma masses suggests a functional role of CEACAM1-integrin beta(3) interaction in melanoma invasion. Moreover, colocalization of the two adhesion molecules is also found at the apical surface of glandular cells of pregnancy endometrium. Colocalization of CEACAM1 and integrin beta(3) at the transitional zone from proliferative to invasive extravillous trophoblast of the maternal fetal interface supports a role for CEACAM1/integrin beta(3) complexes in cell invasion. PMID- 11485913 TI - Proliferation and remodeling of the peritubular microcirculation after nephron reduction: association with the progression of renal lesions. AB - Little is known about the serial changes that might occur in renal capillaries after reduction of renal mass. In the current study, our aim was to document potential alterations in the morphology and proliferation of the renal cortical peritubular microcirculation at specific time points (7 and 60 days) after experimental 75% surgical nephron reduction using two strains of mice that we here demonstrate react differently to the same initial insult: one strain (C57BL6xDBA2/F1 mice) undergoes compensatory growth alone, whereas the other (FVB/N mice) additionally develops severe tubulo-interstitial lesions. Our data demonstrate that significant remodeling and proliferation occur in renal cortical peritubular capillaries after experimental nephron reduction, as assessed by microangiography using infusion of fluorescein isothiocyanate-labeled dextran, expression of the endothelial markers CD34 and Tie-2, and co-expression of CD34 and proliferating cell nuclear antigen, a surrogate marker of cell proliferation. This was accompanied by an increase of renal vascular endothelial growth factor protein levels and a change in distribution of this protein within the kidney itself. Moreover, most of these responses were accentuated in FVB/N mice in the presence of progressive renal disease and positively correlated with tubular epithelial cell proliferation. Hence, we have made three significant novel observations that illuminate the complex pathophysiology of chronic kidney damage after nephron reduction: 1) cortical peritubular capillaries grow by proliferation and remodeling, 2) vascular endothelial growth factor expression is altered, and 3) the development of tubulo-interstitial disease is genetically determined. PMID- 11485914 TI - Proteolysis of AA amyloid fibril proteins by matrix metalloproteinases-1, -2, and -3. AB - We recently demonstrated the presence of matrix metalloproteinases (MMPs)-1, -2, and -3 in AA amyloid deposits, which lead us to speculate that MMPs may participate in amyloidogenesis by either processing the precursor protein, or by degrading the amyloid deposits. Here we investigated this theory by determining the ability of MMP-1, -2, and -3 to degrade human acute-phase serum amyloid A (SAA) and human AA amyloid fibril proteins (AFPs). The following in vitro degradation experiments were performed: using either recombinant MMP-1, -2, or -3 and SAA as a substrate; using either recombinant MMP-1, -2, or -3 and AFP as a substrate; and using THP-1 cells as the protease source and AFP as the substrate. All three MMPs were able to cleave SAA and AFP within the region spanning residues 51 to 57. The following cleavage sites were identified: at 57 to 58 for MMP-1; at 7 to 8 and 51 to 52 for MMP-2; at 7 to 8, 16 to 17, 23 to 24, 51 to 52, 55 to 56, 56 to 57, and 57 to 58 for MMP-3. Cell culture experiments showed that THP-1 cells were able to degrade AFPs. Degradation was significantly delayed after addition of a general metalloproteinase inhibitor (o-phenanthroline) to dextran sulfate-stimulated cells. This is the first study to show that human SAAs and AFPs are susceptible to proteolytic cleavage by MMPs. Immunocytochemistry and electron microscopy showed that degradation takes place in the pericellular or extracellular compartment. PMID- 11485915 TI - No significant association of Epstein-Barr virus infection with invasive breast carcinoma. AB - We studied 48 cases of invasive breast carcinoma for evidence of Epstein-Barr virus (EBV), which is associated with many human malignancies. In situ hybridization studies to detect the presence of EBV-encoded small nonpolyadenylated RNA (EBER)-1 were performed in paraffin sections. Immunohistochemical studies to detect EBV nuclear antigen (EBNA)-1, latent membrane protein (LMP)-1, and the transactivating immediate-early BZLF1 (ZEBRA) protein were also performed in paraffin sections. The presence of EBV genomic DNA was studied by polymerase chain reaction (PCR) amplification using sets of primers flanking the EBNA-4 and the EBV-LMP-1 genes in frozen tissues. Southern blot analysis using a probe flanking the EBV terminal repeat region was then attempted in cases that were PCR-positive. Five of 48 cases (10%) of breast carcinoma showed focal EBER-positive tumor cells. Twelve cases (25%) were positive for EBNA-1 by immunohistochemistry, all but one different from the EBER positive cases. None of the cases were positive for LMP-1 or ZEBRA protein by immunohistochemistry. PCR studies for EBNA-4 and LMP-1 were each positive in five cases (including three cases in common). However, Southern blot studies successfully performed in all but one of the PCR-positive cases were completely negative. The identification of EBV by any methodology was not correlated with tumor size, grade, or lymph node status. This study demonstrated evidence of EBV infection in tissues involved by invasive breast carcinomas in a significant subset of cases. However, the lack of localization of EBV infection to a significant population of the tumor cells in any case, the negativity by Southern blot hybridization, and the lack of expression of multiple antigens in any case strongly argue against a significant role for EBV in the pathogenesis of breast carcinoma. PMID- 11485916 TI - Normal and malignant prostate epithelial cells differ in their response to hepatocyte growth factor/scatter factor. AB - Hepatocyte growth factor/scatter factor (HGF/SF) promotes the proliferation, differentiation, motility, and invasion of epithelial cells by binding to its cell surface receptor, the Met tyrosine kinase. In the prostate, Met is expressed predominantly by prostate epithelial cells (PrEC), whereas HGF/SF is synthesized by prostate stromal cells (PrSC). Met is also expressed in localized and metastatic prostate cancers. Our results show that PrECs in in vitro culture maintain expression of Met at a level comparable to DU145 cancer cell expression. HGF/SF secreted by PrSC stimulates tyrosine phosphorylation of the Met receptor. In normal PrEC, HGF/SF causes growth inhibition, sustained phosphorylation of mitogen-activated protein kinase, and increased CK18 expression consistent with cell differentiation. In contrast, HGF/SF significantly stimulates the proliferation of DU145 prostate cancer cells. HGF/SF in the conditioned medium of PrSC specifically induces migration of both normal and malignant prostate epithelial cells through MatriGel-coated Transwell filters. HGF/SF depletion reduces cell migration by approximately 50%. The response of PrEC is specific for HGF/SF since the other growth factors tested do not significantly affect growth or migration of PrECs. These results support the in vivo importance of the prostate stroma and specifically of HGF/SF as a unique stromal derived factor in the development and progression of prostate cancer. PMID- 11485917 TI - Role of peroxisome proliferator-activated receptor gamma and its ligands in non neoplastic and neoplastic human urothelial cells. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily of ligand-activated transcription factors and is expressed in several types of tissue. Although PPARgamma reportedly is expressed in normal urothelium, its function is unknown. We examined the expression of PPARgamma in normal urothelium and bladder cancer in an attempt to assess its functional role. Immunohistochemical staining revealed normal urothelium to express PPARgamma uniformly. All low-grade carcinomas were positive either diffusely or focally, whereas staining was primarily focal or absent in high grade carcinomas. A nonneoplastic urothelial cell line (1T-1), a low-grade (RT4) carcinoma cell line, and two high-grade (T24 and 253J) carcinoma cell lines in culture expressed PPARgamma mRNA and protein. Luciferase assay indicated that PPARgamma was functional. PPARgamma ligands (15-deoxy-Delta(12,14)-prostaglandin J(2), troglitazone and pioglitazone) suppressed the growth of nonneoplastic and neoplastic urothelial cells in a dose-dependent manner. However, neoplastic cells were more resistant than were nonneoplastic cells. Failure to express PPARgamma or ineffective transcriptional activity may be some of the mechanisms responsible for resistance to the inhibitory action of PPARgamma ligands. PMID- 11485918 TI - Vascular endothelial growth factor enhances glomerular capillary repair and accelerates resolution of experimentally induced glomerulonephritis. AB - Vascular endothelial growth factor (VEGF) regulates angiogenesis through endothelial cell proliferation and plays an important role in capillary repair in damaged glomeruli. We tested the hypothesis that VEGF might be beneficial in rats with severe glomerular injury in glomerulonephritis (GN) based on its angiogenic and vascular remodeling properties. Acute GN with severe glomerular destruction was induced in rats by injection of anti-Thy-1.1 antibody (day 0) and Habu-snake venom (day 1). Rats were intraperitoneally injected with recombinant human VEGF(165) (10 microg/100 g body wt/day) or vehicle from day 2 to day 9, and monitored changes in glomerular capillaries, development of glomerular inflammation, and progression to glomerular sclerosis after acute glomerular destruction in both groups. Rats that received anti-Thy-1.1 antibody and Habu snake venom showed severe mesangiolysis and marked destruction of capillary network on day 2. VEGF was expressed on glomerular epithelial cells, proliferating mesangial cells, and some infiltrating leukocytes, and VEGF(165) protein levels increased in damaged glomeruli during day 5 to day 7. Normal, damaged, and regenerating glomerular endothelial cells expressed VEGF receptor flk-1. However, endothelial cell proliferation and capillary repair was rare in vehicle-treated rats with severe glomerular damage, which progressed to global sclerosis and chronic renal failure by week 8. In contrast, in the VEGF-treated group, VEGF(165) significantly enhanced endothelial cell proliferation and capillary repair in glomeruli by day 9 (proliferating endothelial cells: VEGF(165), 4.3 +/- 1.1; control, 2.2 +/- 0.9 cells on day 7, P < 0.001; and glomerular capillaries: VEGF(165), 24.6 +/- 4.8; control, 16.9 +/- 3.4 capillaries on day 7, P < 0.01). Thereafter, damaged glomeruli gradually recovered after development of capillary network by week 8, and significant improvement of renal function was evident in the VEGF-treated group during week 8 (creatinine: VEGF(165), 0.3 +/- 0.1; control, 2.6 +/- 0.9 mg/dl, P < 0.001; proteinuria: VEGF(165), 54 +/- 15; control, 318 +/- 60 mg/day, P < 0.001). We conclude that the beneficial effect of VEGF(165) in severe glomerular injury in GN emphasizes the importance of capillary repair in the resolution of GN, and may allow the design of new therapeutic strategies against severe GN. PMID- 11485919 TI - SPARC (secreted protein acidic and rich in cysteine) induces apoptosis in ovarian cancer cells. AB - Secreted protein acidic and rich in cysteine (SPARC) is an extracellular Ca(2+) binding matricellular glycoprotein that associates with cell populations undergoing migration, morphogenesis, and differentiation. Studies on endothelial cells have established that its principal functions in vitro are counteradhesion and antiproliferation. The mechanism(s) underlying these antitumor effects is unknown. In this study, we showed that SPARC expression in ovarian cancer cells is inversely correlated with the degree of malignancy. The immunohistochemical data presented here confirmed the importance of diminished SPARC expression in ovarian cancer development. Treating human ovarian surface epithelial cells and ovarian cancer cells with SPARC revealed that as SPARC inhibits the proliferation of both normal and cancer cells, it induces apoptosis only in cancer cells. This observation indicates that down-regulation of SPARC is essential for ovarian carcinogenesis as cancer cells become sensitized to the apoptotic activity of SPARC during malignant transformation. We also showed here the first direct evidence that putative SPARC receptors are present on ovarian epithelial cells. Their levels are higher in human ovarian surface epithelial cells than cancer cells. Binding of SPARC to its receptor is likely to trigger tissue-specific signaling pathways that mediate its tumor suppressing functions. Decrease in ligand-receptor interaction by the down-regulation of SPARC and/or its receptor is essential for ovarian carcinogenesis. PMID- 11485920 TI - Adenomyosis--a result of disordered stromal differentiation. AB - Adenomyosis is a fairly frequent disorder in adult women characterized by the haphazard location of endometrial glands and stroma deep within the myometrium of the uterus. This study compared the effects on uterine development of the selective estrogen receptor modulators, tamoxifen, toremifene, and raloxifene with estradiol when given orally to female mice on days 2 to 5 after birth. Uterine adenomyosis was found in all (14 of 14) mice dosed with tamoxifen and most mice (12 of 14) treated with toremifene, but in none of the vehicle-dosed controls, in only one animal treated with raloxifene at 42 and 90 days after dosing and in none of the mice treated with estradiol at 42 days. At 6 days, the uterus in the groups that developed a high incidence of adenomyosis showed histological evidence of disturbed differentiation of the myometrium. Gene expression XY-scatterplots using Clontech mouse 1.2 Atlas mouse cDNA expression arrays analyzing total uterine RNA showed nerve growth factor-alpha, preadipocyte factor-1, and insulin-like growth factor-2 were key genes differentially modified by tamoxifen or toremifene treatment, relative to the controls. As these genes may play an important role in regulating differentiation and development of the myometrium, these data suggest that adenomyosis may be caused primarily by defects in the formation of the myometrium. PMID- 11485921 TI - Role of interleukin-12 and stat-4 in the regulation of airway inflammation and hyperreactivity in respiratory syncytial virus infection. AB - Respiratory syncytial virus (RSV) is a respiratory pathogen that can cause significant morbidity in infants and young children. Interestingly, the majority of children who acquire a RSV infection do not exhibit severe symptoms. Development of a Th1 response has been associated with resolution of symptoms in viral infections and may explain mild RSV illness. The current study investigated the cytokine response observed in mild disease in C57BL/6 mice that had low airway resistance and mucus production with little pulmonary inflammation. RSV infection in these mice was accompanied by a fourfold increase in interleukin 12(IL-12). Treatment of RSV-infected mice with anti-IL-12 resulted in an increase in airway hyperreactivity, mucus production, and airway inflammation (eosinophilia). Since IL-12 activation is dependent on Stat-4-mediated intracellular signal transduction, similar experiments were performed in Stat-4 deficient mice and demonstrated similar results to those obtained from anti-IL-12 treated mice. Again, there was an increase in airway hyperreactivity and mucus production, and goblet cell hypertrophy. These studies support the importance of IL-12 in the immune response to RSV infection resulting in resolution of disease and protection from inappropriate inflammatory responses. PMID- 11485923 TI - Vascular endothelial growth factor-mediated autocrine stimulation of prostate tumor cells coincides with progression to a malignant phenotype. AB - Vascular endothelial growth factor (VEGF), which is often produced at high levels by tumor cells, is a well-known mediator of tumor angiogenesis. VEGF receptor tyrosine kinases, KDR/Flk-1 and Flt-1, have been thought to be expressed exclusively by endothelial cells. In this study, we have used a prostate tumor progression series comprised of a differentiated rat prostate epithelial cell line, NbE-1, and its highly motile clonal derivative, FB2. Injection of NbE-1 cells into the inferior vena cava of syngeneic rats indicated that these cells are nontumorigenic. Using the same model, FB2 cells generated rapidly growing and well-vascularized tumors in the lungs. NbE-1 expressed marginal levels of VEGF, whereas high levels of VEGF protein were detected in FB2-conditioned medium and in FB2 tumors in vivo. Analysis of (125)I-VEGF(165) binding to NbE-1 and FB2 cells indicated that only motile FB2 cells expressed the VEGF receptor Flt-1. Consistent with this finding, physiological concentrations of VEGF induced chemotactic migration in FB2 but not in NbE-1 cells. This is the first documentation of a functional Flt-1 receptor in prostate tumor cells. Our results suggest two roles for VEGF in tumor progression: a paracrine role as an angiogenic factor and a previously undescribed role as an autocrine mediator of tumor cell motility. PMID- 11485922 TI - Transgenic mice with cardiac-specific expression of activating transcription factor 3, a stress-inducible gene, have conduction abnormalities and contractile dysfunction. AB - Activating transcription factor 3 (ATF3) is a member of the CREB/ATF family of transcription factors. Previously, we demonstrated that the expression of the ATF3 gene is induced by many stress signals. In this report, we demonstrate that expression of ATF3 is induced by cardiac ischemia coupled with reperfusion (ischemia-reperfusion) in both cultured cells and an animal model. Transgenic mice expressing ATF3 under the control of the alpha-myosin heavy chain promoter have atrial enlargement, and atrial and ventricular hypertrophy. Microscopic examination showed myocyte degeneration and fibrosis. Functionally, the transgenic heart has reduced contractility and aberrant conduction. Interestingly, expression of sorcin, a gene whose product inhibits the release of calcium from sarcoplasmic reticulum, is increased in these transgenic hearts. Taken together, our results indicate that expression of ATF3, a stress-inducible gene, in the heart leads to altered gene expression and impaired cardiac function. PMID- 11485924 TI - Alterations of the tumor suppressor genes CDKN2A (p16(INK4a)), p14(ARF), CDKN2B (p15(INK4b)), and CDKN2C (p18(INK4c)) in atypical and anaplastic meningiomas. AB - We investigated 67 meningothelial tumors (20 benign meningiomas, 34 atypical meningiomas, and 13 anaplastic meningiomas) for losses of genetic information from chromosome arms 1p and 9p, as well as for deletion, mutation, and expression of the tumor suppressor genes CDKN2A (p16(INKa)/MTS1), p14(ARF), CDKN2B (p15(INK4b)/MTS2) (all located at 9p21) and CDKN2C (1p32). Comparative genomic hybridization and microsatellite analysis showed losses on 1p in 11 anaplastic meningiomas (85%), 23 atypical meningiomas (68%), and 5 benign meningiomas (25%). One atypical meningioma with loss of heterozygosity on 1p carried a somatic CDKN2C mutation (c.202C>T: R68X). Losses on 9p were found in five anaplastic meningiomas (38%), six atypical meningiomas (18%), and one benign meningioma (5%). Six anaplastic meningiomas (46%) and one atypical meningioma (3%) showed homozygous deletions of the CDKN2A, p14(ARF), and CDKN2B genes. Two anaplastic meningiomas carried somatic point mutations in CDKN2A (c.262G>T: E88X and c.262G>A: E88K) and p14(ARF) (c.305G>T: G102V and c.305G>A: G102E). One anaplastic meningioma, three atypical meningiomas, and one benign meningioma without a demonstrated homozygous deletion or mutation of CDKN2A, p14(ARF), or CDKN2B lacked detectable transcripts from at least one of these genes. Hypermethylation of CDKN2A, p14(ARF), and CDKN2B could be demonstrated in one of these cases. Taken together, our results indicate that CDKN2C is rarely altered in meningiomas. However, the majority of anaplastic meningiomas either show homozygous deletions of CDKN2A, p14(ARF), and CDKN2B, mutations in CDKN2A and p14(ARF), or lack of expression of one or more of these genes. Thus, inactivation of the G(1)/S-phase cell-cycle checkpoint is an important aberration in anaplastic meningiomas. PMID- 11485925 TI - Lymphocyte migration to inflamed lacrimal glands is mediated by vascular cell adhesion molecule-1/alpha(4)beta(1) integrin, peripheral node addressin/l selectin, and lymphocyte function-associated antigen-1 adhesion pathways. AB - Sjogren's syndrome is an autoimmune disease characterized by inflammation and destruction of lacrimal and salivary glands. The development of the inflammation requires the migration of lymphocytes from the blood into these tissues. This migration involves multistep cascades with binding of lacrimal gland endothelial adhesion molecules to their ligands on circulating lymphocytes. We used nonobese diabetic mice, which develop autoimmune-mediated lacrimal gland inflammation, as an experimental model to define the adhesion molecules that control lymphocyte migration into inflamed lacrimal glands. We found that vascular endothelia in inflamed areas of lacrimal gland expressed vascular cell adhesion molecule (VCAM) 1 and the peripheral node addressin (PNAd), but not mucosal addressin cell adhesion molecule-1. Most lymphocytes in the inflamed glands expressed alpha(4) integrin, L-selectin, and lymphocyte function-associated antigen (LFA)-1. In vivo studies revealed that antibodies against VCAM-1, alpha(4) integrin, PNAd, L selectin, or LFA-1 almost completely blocked lymphocyte migration from blood into inflamed lacrimal glands. There was no inhibition of migration by antibodies against mucosal addressin cell adhesion molecule-1 or alpha(4)beta(7) integrin. These results indicate that endothelial/lymphocyte adhesion cascades involving VCAM-1/alpha(4)beta(1) integrin, PNAd/L-selectin, and LFA-1 control the migration of lymphocytes into inflamed lacrimal gland. These adhesion molecules offer potential therapeutic targets to block the development of lacrimal gland inflammation and destruction. PMID- 11485926 TI - Dynamic process of apoptosis in adult rat cardiomyocytes analyzed using 48-hour videomicroscopy and electron microscopy: beating and rate are associated with the apoptotic process. AB - Dynamic process of apoptosis has not been elucidated in adult rat cardiomyocytes. Soluble Fas ligand (0.1 microg/ml) in the presence of actinomycin D (0.05 microg/ml) induced apoptosis in cultured adult rat cardiomyocytes, as documented by activated caspase-3, DNA fragmentation, and apoptotic ultrastructure. In the present model, we observed 60 adult cardiomyocytes with a normal rod shape under a real-time videomicroscope continuously for 48 hours. Seventeen cells (28%) were unchanged and 7 cells (12%) showed oncosis (so-called necrosis) in which no beating was evident. In the remaining 36 cells (apoptosis, 60%), a slow beating (17 +/- 3/min) was initiated 16 +/- 1 hours later. Approximately 1 hour later, the rod cells showed long-axial shortening as bone- or club-like, or square shaped, accompanied with faster beating rates (35 +/- 7/min). In 29 cells (type A1 and A2), marked shrinkage occurred; the cellular shape became almost completely round with a smooth surface and the beating ceased 3.0 +/- 0.4 hours later. Then, smooth budding appeared 0.6 +/- 0.2 hours later. Apoptotic bodies were found in 8 cells 10 +/- 4 hours later (type A1, 13%) but not in 21 cells (type A2, 35%). In the other 7 cells (type A3, 12%), the cell surface became rough 8 +/- 3 hours later and the beating ceased. Maximal beating rate was greatest in type A1 (72 +/- 26/min) and greater in type A2 (29 +/- 5/min) than in type A3 (10 +/- 2/min). Electron microscopy confirmed apoptotic ultrastructure even in the cardiomyocytes with bone-, club-like, or square shapes, suggesting that type A3 as well as A1 and A2 is also under apoptotic process. A caspase inhibitor, zVAD.fmk, blocked beating, apoptotic morphology, and DNA fragmentation, indicating these depended on caspase activation. In the caspase dependent apoptotic process of cultured adult cardiomyocytes, beating and the following deformity of the cellular edges were the initial signs and the rate of beating was related with the subsequent three different processes of apoptosis. PMID- 11485927 TI - Induction of disseminated Mycobacterium avium in simian AIDS is dependent upon simian immunodeficiency virus strain and defective granuloma formation. AB - Mycobacterium avium complex (MAC) is the most common disseminated bacterial disease in patients infected by the human immunodeficiency virus. Although murine models of disseminated MAC exist, they are primarily based on underlying genetic susceptibilities and cannot adequately address the complex interactions that occur between host, mycobacteria, and immunosuppressive lentivirus. To address this problem we have developed an experimental system to co-inoculate rhesus macaques with the simian immunodeficiency virus (SIV) and a clinical M. avium isolate that results in a disease virtually identical to that observed in human cases. Using this experimental system we have found that the development of disseminated MAC is dependent on viral strain. Animals co-infected with SIVmac251 and M. avium developed progressive disease, whereas control animals and animals inoculated with closely related viruses (SIVmac239 and SIVmac239MER) developed self-limiting infections. The ability of animals infected with SIVmac239 or SIVmac239MER to eliminate mycobacterial disease was independent of viral load and CD4 T-cell number but was correlated with the size and composition of microgranulomas. This work establishes a novel primate model of disseminated MAC in the context of immunosuppressive lentiviral infection and advances our understanding of why human immunodeficiency virus-infected patients are remarkably sensitive to the development of mycobacterial disease. PMID- 11485928 TI - Modulation of chagasic cardiomyopathy by interleukin-4: dissociation between inflammation and tissue parasitism. AB - Chronic chagasic cardiomyopathy (CChC) is characterized by an inflammatory reaction which may eventually lead to heart enlargement, arrythmia, and death. As described herein, interleukin-4-deficient mice mount increased specific T helper (Th) 1 immune responses when infected with Trypanosoma cruzi, as compared to wild type mice. Interestingly, these mice had reduced parasitism and mortality and exacerbated inflammation in their hearts, demonstrating a clear dissociation between inflammation and parasite load. The modulation of these phenomena so as to maximize host and parasite survivals may depend on a fine balance between Th responses, in which a Th1 response will, on one hand, control parasitism and, on the other hand, enhance heart inflammation throughout the course of the infection. PMID- 11485929 TI - Down-regulation of telomerase activity in malignant lymphomas by radiation and chemotherapeutic agents. AB - The effects of radiation and cytotoxic agents on telomerase activity in lymphoma cells were analyzed by a polymerase chain reaction-based telomeric repeat amplification protocol coupled with an enzyme-linked immunosorbent assay, reverse transcriptase-polymerase chain reaction for the expression of the catalytic subunit of telomerase (hTERT), and by Western blot analysis in three lymphoma cell lines (Jurkat, Raji, CEM-6). Telomeric repeat amplification protocol-enzyme linked immunosorbent assay demonstrated high basal levels of telomerase activity in all cell lines compared to normal and activated peripheral blood lymphocytes. A significant decrease in telomerase activity was observed in all cell lines after exposure to vincristine for 24 hours. The decrease in telomerase activity paralleled the decrease in cell viability in Jurkat and CEM-6 cells but not in Raji cells. Radiation exposure inhibited the telomerase activity of Jurkat and CEM-6 cells whereas Raji cells were unaffected. Cell cycle analysis demonstrated a significant G(2)/M arrest by cisplatin, VP-16, and vincristine. In contrast to the decline in telomerase activity, the level of hTERT RNA and protein increased. Furthermore, the induction of hTERT was preceded by increased expression of the cyclin-dependent kinase inhibitor, p27/Kip1 protein, and p53. These results indicate that telomerase activity is down-regulated by anti-neoplastic agents in lymphoma cells, however expression of hTERT may not be correlated with telomerase activity. We also show that p27/Kip1 may be involved in the G(2)/M growth arrest induced by anti-neoplastic agents. PMID- 11485930 TI - Methionine aminopeptidase-2 regulates human mesothelioma cell survival: role of Bcl-2 expression and telomerase activity. AB - Methionine aminopeptidase-2 (MetAP2) is the molecular target of the angiogenesis inhibitors, fumagillin and ovalacin. Fumagillin can also inhibit cancer cell proliferation, implying that MetAP2 may play a quite complex role in tumor progression. Here, we examined the expression and function of MetAP2 in an in vitro model of human mesothelioma. We found that mesothelioma cells expressed higher MetAP2 mRNA levels than primary normal mesothelial cells. Consistently, fumagillin induced apoptosis, owing to early mitochondrial damage, in malignant, but not in normal mesothelial cells. Transfection of mesothelioma cells with a MetAP2 anti-sense oligonucleotide determined a time-dependent inhibition of cell survival and induced nucleosome formation. Interestingly, mRNA and protein levels of the anti-apoptotic gene bcl-2 as well as telomerase activity were selectively reduced after MetAP2 inhibition in mesothelioma cells, whereas bcl-2 overexpression counteracted the effect of MetAP2 inhibition on telomerase activity and apoptosis. MetAP2 inhibition also increased caspase activity and the caspase inhibitor, zVAD-fmk, prevented fumagillin-induced apoptosis, but it did not alter telomerase activity. These results indicate that MetAP2 is a main regulator of proliferative and apoptotic pathways in mesothelioma cells and suggest that MetAP2 inhibition may represent a potential target for therapeutic intervention in human mesothelioma. PMID- 11485931 TI - Role of CD40-CVD40L in mouse severe malaria. AB - We explored the role of CD40-CD40L (CD154) in the severe malaria elicited by Plasmodium berghei anka infection in mice. Mortality was >90% by day 8 after infection in +/+ mice, but markedly decreased in CD40-/- or in CD40L-/- mice, as well as in +/+ mice treated with anti-CD40L monoclonal antibody. Parasitemia was similar in the different conditions. Breakdown of the blood-brain barrier was evident in infected +/+, but not in CD40-/- mice. Thrombocytopenia was less severe in CD40-/- mice than in the +/+ controls. Sequestration of macrophages in brain venules and alveolar capillaries was reduced in CD40-/- or in CD40L-/- mice, whereas sequestration of parasitized red blood cells or polymorphonuclear leukocytes in alveolar capillaries was CD40-CD40L-independent. CD40 mRNA was increased in the brain and lung of infected mice whereas CD40L was increased in the lung. Tumor necrosis factor plasma levels were similarly increased in infected +/+ or CD40-/- mice. Expression of CD54 and its mRNA levels in the brain were moderately decreased in CD40-deficient mice. Thus the mortality associated with severe malaria requires CD40-CD40L interaction that contributes to the breakdown of the blood-brain barrier, macrophage sequestration, and platelet consumption. PMID- 11485932 TI - Renal cholesterol accumulation: a durable response after acute and subacute renal insults. AB - Proximal tubular cholesterol levels rise within 18 hours of diverse forms of acute renal tubular injury (eg, myoglobinuria, ischemia/reperfusion, urinary tract obstruction). These increments serve to protect against further bouts of tubular attack (so-called "acquired cytoresistance"). Whether these cholesterol increments are merely transitory, or persist into the maintenance phase of acute renal failure (ARF), has not been previously defined. Furthermore, whether subacute/insidious tubular injury [eg, cyclosporine A (CSA), tacrolimus toxicity], nontubular injury (eg, acute glomerulonephritis), or physiological stress (eg, mild dehydration) impact renal cholesterol homeostasis have not been addressed. This study sought to resolve these issues. Male CD-1 mice were subjected to glycerol-induced ARF. Renal cortical-free cholesterol (FC) and cholesterol ester (CE) levels were determined 3, 5, 7, or 14 days later, and the values contrasted to prevailing blood-urea nitrogen concentrations. The impact of 40 minutes of unilateral renal ischemia plus reflow (3 to 6 days) on mouse cortical FC/CE content was also assessed. Additionally, FC/CE levels were measured in rat renal cortex either 10 days after CSA or tacrolimus therapy, or 48 hours after induction of nephrotoxic serum nephritis. Finally, the impact of overnight dehydration on mouse renal cortical/medullary FC/CE profiles was determined. Compared to sham-treated animals, glycerol, CSA, tacrolimus, ischemia reperfusion, and nephrotoxic serum each induced dramatic CE +/- FC elevations, rising as much as 10x control values. In the glycerol model, striking correlations (r 500 microg/day were limited. The dose response curve for the raw data began to reach a plateau at around 100-200 microg/day and peaked by 500 microg/day. A negative exponential model for the data, without meta-analysis, indicated that 80% of the benefit at 1000 microg/day was achieved at doses of 70-170 microg/day and 90% by 100-250 microg/day. A quadratic meta-regression showed that the maximum achievable efficacy was obtained by doses of around 500 microg/day. The odds ratio for patients remaining in a study at a dose of 200 microg/day, compared with higher doses, was 0.73 (95% confidence interval 0.49 to 1.08). Comparison of the standardised difference in FEV(1 )for an inhaled dose of 200 microg/day against higher doses showed a difference in FEV(1) of 0.13 of a standard deviation (-0.02 to 0.29). CONCLUSIONS: In adolescent and adult patients with asthma, most of the therapeutic benefit of inhaled fluticasone is achieved with a total daily dose of 100-250 microg, and the maximum effect is achieved with a dose of around 500 microg/day. However, these findings were limited by the lack of data on individual patients and by the paucity of dose-response studies that included doses of >500 microg/day. PMID- 11485953 TI - Cohort study of depressed mood during pregnancy and after childbirth. AB - OBJECTIVE: To follow mothers' mood through pregnancy and after childbirth and compare reported symptoms of depression at each stage. DESIGN: Longitudinal cohort study. SETTING: Avon. PARTICIPANTS: Pregnant women resident within Avon with an expected date of delivery between 1 April 1991 and 31 December 1992. MAIN OUTCOME MEASURES: Symptom scores from the Edinburgh postnatal depression scale at 18 and 32 weeks of pregnancy and 8 weeks and 8 months postpartum. Proportion of women above a threshold indicating probable depressive disorder. RESULTS: Depression scores were higher at 32 weeks of pregnancy than 8 weeks postpartum (difference in means 0.88, 95% confidence interval 0.79 to 0.97). There was no difference in the distribution of total scores or scores for individual items at the four time points. 1222 (13.5%) women scored above threshold for probable depression at 32 weeks of pregnancy, 821 (9.1%) at 8 weeks postpartum, and 147 (1.6%) throughout. More mothers moved above the threshold for depression between 18 weeks and 32 weeks of pregnancy than between 32 weeks of pregnancy and 8 weeks postpartum. CONCLUSIONS: Symptoms of depression are not more common or severe after childbirth than during pregnancy. Research and clinical efforts need to be moved towards understanding, recognising, and treating antenatal depression. PMID- 11485954 TI - Impact of published clinical outcomes data: case study in NHS hospital trusts. AB - OBJECTIVE: To examine the impact of the publication of clinical outcomes data on NHS Trusts in Scotland to inform the development of similar schemes elsewhere. DESIGN: Case studies including semistructured interviews and a review of background statistics. SETTING: Eight Scottish NHS acute trusts. PARTICIPANTS: 48 trust staff comprising chief executives, medical directors, stroke consultants, breast cancer consultants, nurse managers, and junior doctors. MAIN OUTCOME MEASURES: Staff views on the benefits and drawbacks of clinical outcome indicators provided by the clinical resource and audit group (CRAG) and perceptions of the impact of these data on clinical practice and continuous improvement of quality. RESULTS: The CRAG indicators had a low profile in the trusts and were rarely cited as informing internal quality improvement or used externally to identify best practice. The indicators were mainly used to support applications for further funding and service development. The poor effect was attributable to a lack of professional belief in the indicators, arising from perceived problems around quality of data and time lag between collection and presentation of data; limited dissemination; weak incentives to take action; a predilection for process rather than outcome indicators; and a belief that informal information is often more useful than quantitative data in the assessment of clinical performance. CONCLUSIONS: Those responsible for developing clinical indicator programmes should develop robust datasets. They should also encourage a working environment and incentives such that these data are used to improve continuously. PMID- 11485955 TI - Declaring financial competing interests: survey of five general medical journals. PMID- 11485956 TI - Effect of Helicobacter pylori infection on blood pressure: a community based cross sectional study. PMID- 11485957 TI - How should hamsters run? Some observations about sufficient patient time in primary care. PMID- 11485958 TI - General practice workload implications of the national service framework for coronary heart disease: cross sectional survey. PMID- 11485960 TI - Developing strategies for long term follow up of survivors of childhood cancer. PMID- 11485961 TI - Evidence based policy: proceed with care. PMID- 11485968 TI - Drilling for oxygen: angiogenesis involves proteolysis of the extracellular matrix. PMID- 11485969 TI - Oxidative stress and apoptosis in heart failure progression. PMID- 11485970 TI - Matrix metalloproteinase inhibition after myocardial infarction: a new approach to prevent heart failure? AB - Increased activity of matrix metalloproteinases (MMPs) has been implicated in numerous disease processes, including tumor growth and metastasis, arthritis, and periodontal disease. It is now becoming increasingly clear that extracellular matrix degradation by MMPs is also involved in the pathogenesis of cardiovascular disease, including atherosclerosis, restenosis, dilated cardiomyopathy, and myocardial infarction. Administration of synthetic MMP inhibitors in experimental animal models of these cardiovascular diseases significantly inhibits the progression of, respectively, atherosclerotic lesion formation, neointima formation, left ventricular remodeling, pump dysfunction, and infarct healing. This review focuses on the role of MMPs in cardiovascular disease, in particular myocardial infarction and the subsequent progression to heart failure. MMPs, which are present in the myocardium and capable of degrading all the matrix components of the heart, are the driving force behind myocardial matrix remodeling. The recent finding that acute pharmacological inhibition of MMPs or deficiency in MMP-9 attenuates left ventricular dilatation in the infarcted mouse heart led to the proposal that MMP inhibitors could be used as a potential therapy for patients at risk for the development of heart failure after myocardial infarction. Although these promising results encourage the design of clinical trials with MMP inhibitors, there are still several unresolved issues. This review describes the biology of MMPs and discusses new insights into the role of MMPs in several cardiovascular diseases. Attention will be paid to the central role of the plasminogen system as an important activator of MMPs in the remodeling process after myocardial infarction. Finally, we speculate on the use of MMP inhibitors as potential therapy for heart failure. PMID- 11485971 TI - Functional consequences of integrin gene mutations in mice. AB - Integrins are cell-surface receptors responsible for cell attachment to extracellular matrices and to other cells. The application of mouse genetics has significantly increased our understanding of integrin function in vivo. In this review, we summarize the phenotypes of mice carrying mutant integrin genes and compare them with phenotypes of mice lacking the integrin ligands. PMID- 11485972 TI - Methods of detection of vascular reactive species: nitric oxide, superoxide, hydrogen peroxide, and peroxynitrite. AB - The evanescent nature of reactive oxygen and nitrogen species, the multiple cellular mechanisms evolved to maintain these substances at low (submicromolar) concentrations within the vascular system, and the often multifaceted nature of their reactivities have made measurement of these compounds within the vasculature problematic. This review attempts to provide a critical description of some of the most common approaches to quantification of nitric oxide, superoxide, hydrogen peroxide, and peroxynitrite, with attention to key issues that may influence the utility of a particular assay when adapted for use in vascular cells and tissues. PMID- 11485973 TI - Mechanisms of telomerase induction during vascular smooth muscle cell proliferation. AB - Telomeres are primarily controlled by a highly specialized DNA polymerase termed telomerase. Recent studies have demonstrated that introduction of the telomerase catalytic component (TERT) into telomerase-negative cells activates telomerase and extends cell life span, whereas mice lacking telomerase activity revealed impaired cell proliferation in some organs as well as reduced tumorigenesis. These reports suggest that telomerase plays an important role in long-term cell viability and cell proliferation. However, the mechanism or mechanisms by which telomerase is induced or regulated remains to be elucidated. We report here that primary vascular smooth muscle cells (VSMCs) express telomerase and that increased telomerase activity correlates with cell proliferation. Inhibition of telomerase diminished growth of VSMCs, which suggests a crucial role for telomerase activation in the regulation of VSMC proliferation. We propose a novel model whereby telomerase is first activated in the cytoplasm before cell proliferation, followed by accumulation of activity in the nucleus during the logarithmic phase of cell growth. Activation of telomerase in VSMCs was linked to phosphorylation of TERT. The protein kinase inhibitor H7 suppressed the activation of telomerase in the cytoplasm and also inhibited the accumulation of TERT as well as telomerase activity in the nucleus. These data suggest that posttranslational modification of TERT by phosphorylation is important for activation and accumulation of telomerase into the nucleus in the process of VSMC proliferation. PMID- 11485974 TI - Chlamydia pneumoniae and chlamydial heat shock protein 60 stimulate proliferation of human vascular smooth muscle cells via toll-like receptor 4 and p44/p42 mitogen-activated protein kinase activation. AB - An early component of atherogenesis is abnormal vascular smooth muscle cell (VSMC) proliferation. The presence of Chlamydia pneumoniae in many atherosclerotic lesions raises the possibility that this organism plays a causal role in atherogenesis. In this study, C pneumoniae elementary bodies (EBs) rapidly activated p44/p42 mitogen-activated protein kinases (MAPKs) and stimulated proliferation of VSMCs in vitro. Exposure of VSMCs derived from human saphenous vein to C pneumoniae EBs (3x10(7) inclusion forming units/mL) enhanced bromodeoxyuridine (BrdU) incorporation 12+/-3-fold. UV- and heat-inactivated C pneumoniae EBs also stimulated VSMC proliferation, indicating a role of direct stimulation by chlamydial antigens. However, the mitogenic activity of C pneumoniae was heat-labile, thus excluding a role of lipopolysaccharide. Chlamydial hsp60 (25 microg/mL) replicated the effect of C pneumoniae, stimulating BrdU incorporation 7+/-3-fold. Exposure to C pneumoniae or chlamydial hsp60 rapidly activated p44/p42 MAPK, within 5 to 10 minutes of exposure. In addition, PD98059 and U0126, which are two distinct inhibitors of upstream MAPK kinase 1/2 (MEK1/2), abolished the mitogenic effect of C pneumoniae and chlamydial hsp60. Toll-like receptors (TLRs) act as sensors for microbial antigens and can signal via the p44/p42 MAPK pathway. Human VSMCs were shown to express TLR4 mRNA and protein, and a TLR4 antagonist abolished chlamydial hsp60 induced VSMC proliferation and attenuated C pneumoniae-induced MAPK activation and VSMC proliferation. Together these results indicate that C pneumoniae and chlamydial hsp60 are potent inducers of human VSMC proliferation and that these effects are mediated, at least in part, by rapid TLR4-mediated activation of p44/p42 MAPK. PMID- 11485975 TI - Phenotypic modulation of vascular smooth muscle cells induced by unsaturated lysophosphatidic acids. AB - The phenotypic modulation of vascular smooth muscle cells (VSMCs) from the differentiated state to the dedifferentiated one is critically involved in the development and progression of atherosclerosis. Although many cytokines and growth factors have been reported as atherogenic factors, the critical pathogens for inducing atherosclerosis remain unknown, largely because proper examining systems of them have not been developed. We recently established primary culture systems for visceral SMCs and VSMCs in which both SMCs, when cultured on laminin with insulin-like growth factor-I, show a differentiated phenotype, as indicated by a spindle-like shape, ligand-induced contractility, and a high level of SMC differentiation marker gene expression. In this study, we searched for critical dedifferentiation factors for these SMCs using our culture system. We found that polar lipids extracted from human serum markedly induced VSMC dedifferentiation, and this activity was solely present in the lysophosphatidic acid (LPA) fraction. Among several LPA species detected in human serum lipids, unsaturated LPAs were identified as major contributors to the induction of VSMC dedifferentiation. Signaling and phenotype analyses revealed that unsaturated LPA-induced VSMC dedifferentiation is mediated through the coordinated activation of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase. Thus, this report demonstrates the first finding that unsaturated LPAs, but not saturated LPAs, specifically induce VSMC phenotypic modulation, suggesting that these molecules could function as atherogenic factors. PMID- 11485976 TI - Regulation of matrix metalloproteinase activity in ischemic tissue by interleukin 10: role in ischemia-induced angiogenesis. AB - We have previously shown that deficiency in the anti-inflammatory cytokine interleukin-10 (IL-10) is responsible for enhanced angiogenesis after hindlimb ischemia. This study examined the putative involvement of matrix metalloproteinase (MMP) activation in this process. Ischemia was produced by artery femoral occlusion in both C57BL6 IL-10(+/+) and IL-10(-/-) mice. Angiographic vessel density and laser Doppler perfusion data at day 28 showed significant improvement in ischemic/nonischemic leg ratio by, respectively, 1.8 fold and 1.4-fold in IL-10(-/-) mice compared with IL-10(+/+) mice. This was associated with an increase in vascular endothelial growth factor (VEGF) protein content in the ischemic hindlimb. Three days after ischemia, gelatin zymography showed a significant increase in both pro- and active forms of MMP-2 and MMP-9 in ischemic hindlimbs of IL-10(-/-) mice compared with IL-10(+/+) mice (P<0.01). This increase in MMP activity in IL-10(-/-) mice was completely inhibited by treatment with BB-94 (5 mg/kg IP), a specific MMP inhibitor. Furthermore, increases in both vessel density and blood perfusion indexes at day 28 in IL-10( /-) mice were abolished after treatment with BB-94 (0.78+/-0.06 versus 1.17+/ 0.09 and 0.62+/-0.02 versus 0.88+/-0.04, for vessel density and blood perfusion ratio, respectively, in IL-10(-/-) mice treated with BB-94 versus untreated IL 10(-/-) mice, P<0.05). In contrast, BB-94 treatment did not affect the rise in VEGF protein content. These findings in IL-10(-/-) mice underscore the critical role of MMP activation, in a context of increased VEGF expression, in promoting ischemia-induced angiogenesis. PMID- 11485977 TI - Gene expression and in situ localization of diacylglycerol kinase isozymes in normal and infarcted rat hearts: effects of captopril treatment. AB - Diacylglycerol (DG) kinase (DGK) terminates signaling from DG, which serves as an activator of protein kinase C (PKC), by converting DG to phosphatidic acid. DGK is thus regarded as an attenuator of the PKC activity. In rats, five DGK isozymes have been cloned, but little is known about their role in the heart. In this study, the spatiotemporal expression of DGK isozymes was investigated in rat hearts under a normal condition and after myocardial infarction (MI) by in situ hybridization histochemistry and immunohistochemistry. In normal left ventricular myocardium, DGKalpha, DGKepsilon, and DGKzeta mRNAs were expressed evenly throughout the myocardium, although the DGKalpha expression was very low. In infarcted hearts, the expression of DGKzeta was enhanced in the peripheral zone of the necrotic area and at the border zone 3 and 7 days after MI, and to a lesser extent in the middle layer of the granulation tissue 21 days after MI. The enhanced DGKzeta expression in the infarcted and border areas could be attributed to granulocytes and macrophages. In contrast, the expression of DGKepsilon in the infarcted and border areas was lower than that in the viable left ventricle (LV) throughout the postoperation period. Furthermore, DGKepsilon expression in the viable myocardium 21 days after MI decreased significantly compared with left ventricular myocardium in the sham-operated rats and was completely restored by treatment with captopril. Our results demonstrate that three DGK isozymes are expressed in the heart and that each isozyme might have different functional characteristics in the healing and LV remodeling after MI. PMID- 11485978 TI - Acetylcholine, bradykinin, opioids, and phenylephrine, but not adenosine, trigger preconditioning by generating free radicals and opening mitochondrial K(ATP) channels. AB - It has been assumed that all G(i)-coupled receptors trigger the protective action of preconditioning by means of an identical intracellular signaling pathway. To test this assumption, rabbit hearts were isolated and perfused with Krebs buffer. All hearts were subjected to a 30-minute coronary artery occlusion followed by 120 minutes of reperfusion. Risk area was measured with fluorescent particles and infarct size with triphenyltetrazolium chloride staining. Control hearts showed 29.1+/-2.8% infarction of the risk zone. A 5-minute infusion of acetylcholine (0.55 mmol/L) beginning 15 minutes before the 30-minute occlusion resulted in significant protection (9.2+/-2.7% infarction). This protection could be blocked by administration of 300 micromol/L N-2-mercaptopropionyl glycine (MPG), a free radical scavenger, or by 200 micromol/L 5-hydroxydecanoate (5-HD), a mitochondrial K(ATP) antagonist, for 15 minutes beginning 5 minutes before the acetylcholine infusion (35.2+/-3.9% and 27.8+/-2.4% infarction, respectively). Similar protection was observed with other known triggers, ie, bradykinin (0.4 micromol/L), morphine (0.3 micromol/L), and phenylephrine (0.1 micromol/L), and in each case protection was completely abrogated by either MPG or 5-HD. In contrast, protection by adenosine or its analog N(6)-(2-phenylisopropyl) adenosine could not be blocked by either MPG or 5-HD. Therefore, whereas most of the tested agonists trigger protection by a pathway that requires opening of mitochondrial K(ATP) channels and production of free radicals, the protective action of adenosine is not dependent on either of these steps. Hence, it cannot be assumed that all G(i)-coupled receptors use the same signal transduction pathways to trigger preconditioning. PMID- 11485979 TI - Oxidative stress-mediated cardiac cell death is a major determinant of ventricular dysfunction and failure in dog dilated cardiomyopathy. AB - Cell death has been questioned as a mechanism of ventricular failure. In this report, we tested the hypothesis that apoptotic death of myocytes, endothelial cells, and fibroblasts is implicated in the development of the dilated myopathy induced by ventricular pacing. Accumulation of reactive oxygen products such as nitrotyrosine, potentiation of the oxidative stress response by p66(shc) expression, formation of p53 fragments, release of cytochrome c, and caspase activation were examined to establish whether these events were coupled with apoptotic cell death in the paced dog heart. Myocyte, endothelial cell, and fibroblast apoptosis was detected before indices of severe impairment of cardiac function became apparent. Cell death increased with the duration of pacing, and myocyte death exceeded endothelial cell and fibroblast death throughout. Nitrotyrosine formation and p66(shc) levels progressively increased with pacing and were associated with cell apoptosis. Similarly, p50 (DeltaN) fragments augmented paralleling the degree of cell death in the failing heart. Moreover, cytochrome c release and activation of caspase-9 and -3 increased from 1 to 4 weeks of pacing. In conclusion, cardiac cell death precedes ventricular decompensation and correlates with the time-dependent deterioration of function in this model. Oxidative stress may be critical for activation of apoptosis in the overloaded heart. PMID- 11485980 TI - What is the organization of waves in ventricular fibrillation? PMID- 11485981 TI - Acid prohormone sequence determines size, shape, and docking of secretory vesicles in atrial myocytes. AB - How vesicles are born in the trans-Golgi network and reach their docking sites at the plasma membrane is still largely unknown and is investigated in the present study on live, primary cultured atrial cardiomyocytes. Secretory vesicles (n=422) are visualized by expressing fusion proteins of proatrial natriuretic peptide (proANP) and green fluorescent protein. Myocytes expressing fusion proteins with intact proANP display two populations of fluorescent vesicles with apparent diameters of 120 and 175 nm, moving at a top velocity of 0.3 microm/s. The number of docked vesicles is significantly correlated with the number of mobile vesicles (r=0.71, P<0.0005). The deletion of the acidic N-terminal proANP[1-44] or point mutations (glu(23,24)-->gln(23,24)) change size and shape-but not velocity-of the vesicles, and, strikingly, abolish their docking at the plasma membrane. The shapes thus change from spheres to larger, irregular floppy bags or vesicle trains. Deletion of the C-terminal proANP[45-127], where the ANP and its disulfide bond reside, does not change size, shape, docking, or velocity of the mobile vesicles. The N-terminal acid calcium-binding sequence of proANP is known to cause protein aggregation at the high calcium concentration prevailing in the trans-Golgi network. Therefore, these results indicate that amino acid residues favoring cargo aggregation are critically important in shaping the secretory vesicles and determining their fate-docking or not docking-at the plasma membrane. The full text of this article is available at http://www.circresaha.org. PMID- 11485982 TI - CREating a SCAP-less liver keeps SREBPs pinned in the ER membrane and prevents increased lipid synthesis in response to low cholesterol and high insulin. PMID- 11485983 TI - BMPs on the road to hepatogenesis. PMID- 11485984 TI - Nrarp is a novel intracellular component of the Notch signaling pathway. AB - The Lin12/Notch receptors regulate cell fate during embryogenesis by activating the expression of downstream target genes. These receptors signal via their intracellular domain (ICD), which is released from the plasma membrane by proteolytic processing and associates in the nucleus with the CSL family of DNA binding proteins to form a transcriptional activator. How the CSL/ICD complex activates transcription and how this complex is regulated during development remains poorly understood. Here we describe Nrarp as a new intracellular component of the Notch signaling pathway in Xenopus embryos. Nrarp is a member of the Delta-Notch synexpression group and encodes a small protein containing two ankyrin repeats. Nrarp expression is activated in Xenopus embryos by the CSL dependent Notch pathway. Conversely, overexpression of Nrarp in embryos blocks Notch signaling and inhibits the activation of Notch target genes by ICD. We show that Nrarp forms a ternary complex with the ICD of XNotch1 and the CSL protein XSu(H) and that in embryos Nrarp promotes the loss of ICD. By down-regulating ICD levels, Nrarp could function as a negative feedback regulator of Notch signaling that attenuates ICD-mediated transcription. PMID- 11485985 TI - Mutations in the Drosophila dTAK1 gene reveal a conserved function for MAPKKKs in the control of rel/NF-kappaB-dependent innate immune responses. AB - In mammals, TAK1, a MAPKKK kinase, is implicated in multiple signaling processes, including the regulation of NF-kappaB activity via the IL1-R/TLR pathways. TAK1 function has largely been studied in cultured cells, and its in vivo function is not fully understood. We have isolated null mutations in the Drosophila dTAK1 gene that encodes dTAK1, a homolog of TAK1. dTAK1 mutant flies are viable and fertile, but they do not produce antibacterial peptides and are highly susceptible to Gram-negative bacterial infection. This phenotype is similar to the phenotypes generated by mutations in components of the Drosophila Imd pathway. Our genetic studies also indicate that dTAK1 functions downstream of the Imd protein and upstream of the IKK complex in the Imd pathway that controls the Rel/NF-kappaB like transactivator Relish. In addition, our epistatic analysis places the caspase, Dredd, downstream of the IKK complex, which supports the idea that Relish is processed and activated by a caspase activity. Our genetic demonstration of dTAK1's role in the regulation of Drosophila antimicrobial peptide gene expression suggests an evolutionary conserved role for TAK1 in the activation of Rel/NF-kappaB-mediated host defense reactions. PMID- 11485986 TI - PDGF autocrine stimulation dedifferentiates cultured astrocytes and induces oligodendrogliomas and oligoastrocytomas from neural progenitors and astrocytes in vivo. AB - We present evidence that some low-grade oligodendrogliomas may be comprised of proliferating glial progenitor cells that are blocked in their ability to differentiate, whereas malignant gliomas have additionally acquired other mutations such as disruption of cell cycle arrest pathways by loss of Ink4a-Arf. We have modeled these effects in cell culture and in mice by generating autocrine stimulation of glia through the platelet-derived growth factor receptor (PDGFR). In cell culture, PDGF signaling induces proliferation of glial precursors and blocks their differentiation into oligodendrocytes and astrocytes. In addition, coexpression of PDGF and PDGF receptors has been demonstrated in human gliomas, implying that autocrine stimulation may be involved in glioma formation. In this study, using somatic cell type-specific gene transfer we investigated the functions of PDGF autocrine signaling in gliomagenesis by transferring the overexpression of PDGF-B into either nestin-expressing neural progenitors or glial fibrillary acidic protein (GFAP)-expressing astrocytes both in cell culture and in vivo. In cultured astrocytes, overexpression of PDGF-B caused significant increase in proliferation rate of both astrocytes and neural progenitors. Furthermore, PDGF gene transfer converted cultured astrocytes into cells with morphologic and gene expression characteristics of glial precursors. In vivo, gene transfer of PDGF to neural progenitors induced the formation of oligodendrogliomas in about 60% of mice by 12 wk of age; PDGF transfer to astrocytes induced the formation of either oligodendrogliomas or mixed oligoastrocytomas in about 40% of mice in the same time period. Loss of Ink4a Arf, a mutation frequently found in high-grade human gliomas, resulted in shortened latency and enhanced malignancy of gliomas. The highest percentage of PDGF-induced malignant gliomas arose from of Ink4a-Arf null progenitor cells. These data suggest that chronic autocrine PDGF signaling can promote a proliferating population of glial precursors and is potentially sufficient to induce gliomagenesis. Loss of Ink4a-Arf is not required for PDGF-induced glioma formation but promotes tumor progression toward a more malignant phenotype. PMID- 11485987 TI - Functional inactivation of the IGF-I and insulin receptors in skeletal muscle causes type 2 diabetes. AB - Peripheral insulin resistance and impaired insulin action are the primary characteristics of type 2 diabetes. The first observable defect in this major disorder occurs in muscle, where glucose disposal in response to insulin is impaired. We have developed a transgenic mouse with a dominant-negative insulin like growth factor-I receptor (KR-IGF-IR) specifically targeted to the skeletal muscle. Expression of KR-IGF-IR resulted in the formation of hybrid receptors between the mutant and the endogenous IGF-I and insulin receptors, thereby abrogating the normal function of these receptors and leading to insulin resistance. Pancreatic beta-cell dysfunction developed at a relative early age, resulting in diabetes. These mice provide an excellent model to study the molecular mechanisms underlying the development of human type 2 diabetes. PMID- 11485988 TI - SAGA is an essential in vivo target of the yeast acidic activator Gal4p. AB - Despite major advances in characterizing the eukaryotic transcriptional machinery, the function of promoter-specific transcriptional activators (activators) is still not understood. For example, in no case have the direct in vivo targets of a transcriptional activator been unambiguously identified, nor has it been resolved whether activators have a single essential target or multiple redundant targets. Here we address these issues for the prototype acidic activator yeast Gal4p. Gal4p binds to the upstream activating sequence (UAS) of GAL1 and several other GAL genes and stimulates transcription in the presence of galactose. Previous studies have shown that GAL1 transcription is dependent on the yeast SAGA (Spt/Ada/GCN5/acetyltransferase) complex. Using formaldehyde-based in vivo cross-linking, we show that the Gal4p activation domain recruits SAGA to the GAL1 UAS. If SAGA is not recruited to the UAS, the preinitiation complex (PIC) fails to assemble at the GAL1 core promoter, and transcription does not occur. SAGA, but not other transcription components, is also recruited by the Gal4p activation domain to a plasmid containing minimal Gal4p-binding sites. Recruitment of SAGA by Gal4p and stimulation of PIC assembly is dependent on several SAGA subunits but not the SAGA histone acetyl-transferase (HAT) GCN5. Based on these and other results, we conclude that SAGA is an essential target of Gal4p that, following recruitment to the UAS, facilitates PIC assembly and transcription. PMID- 11485989 TI - The S. cerevisiae SAGA complex functions in vivo as a coactivator for transcriptional activation by Gal4. AB - Previous studies demonstrated that the SAGA (Spt-Ada-Gcn5-Acetyltransferase) complex facilitates the binding of TATA-binding protein (TBP) during transcriptional activation of the GAL1 gene of Saccharomyces cerevisiae. TBP binding was shown to require the SAGA components Spt3 and Spt20/Ada5, but not the SAGA component Gcn5. We have now examined whether SAGA is directly required as a coactivator in vivo by using chromatin immunoprecipitation analysis. Our results demonstrate that SAGA is physically recruited in vivo to the upstream activation sequence (UAS) regions of the galactose-inducible GAL genes. This recruitment is dependent on both induction by galactose and the Gal4 activation domain. Furthermore, we demonstrate that another well-characterized activator, Gal4-VP16, also recruits SAGA in vivo. Finally, we provide evidence that a specific interaction between Spt3 and TBP in vivo is important for Gal4 transcriptional activation at a step after SAGA recruitment. These results, taken together with previous studies, demonstrate a dependent pathway for the recruitment of TBP to GAL gene promoters consisting of the recruitment of SAGA by Gal4 and the subsequent recruitment of TBP by SAGA. PMID- 11485990 TI - Characterization of U4 and U6 interactions with the 5' splice site using a S. cerevisiae in vitro trans-splicing system. AB - Spliceosome assembly has been characterized as the ordered association of the snRNP particles U1, U2, and U4/U6.U5 onto pre-mRNA. We have used an in vitro trans-splicing/cross-linking system in Saccharomyces cerevisiae nuclear extracts to examine the first step of this process, 5' splice site recognition. This trans splicing reaction has ATP, Mg(2+), and splice-site sequence requirements similar to those of cis-splicing reactions. Using this system, we identified and characterized a novel U4-5' splice site interaction that is ATP-dependent, but does not require the branch point, the 3' splice site, or the 5' end of the U1 snRNA. Additionally, we identified several ATP-dependent U6 cross-links at the 5' splice site, indicating that different regions of U6 sample it before a U6-5' splice site interaction is stabilized that persists through the first step of splicing. This work provides evidence for ATP-dependent U4/U6 association with the 5' splice site independent of ATP-mediated U2 association with the branch point. Furthermore, it defines specific nucleotides in U4 and U6 that interact with the 5' splice site at this early stage, even in the absence of base-pairing with the U1 snRNA. PMID- 11485991 TI - FIERY1 encoding an inositol polyphosphate 1-phosphatase is a negative regulator of abscisic acid and stress signaling in Arabidopsis. AB - The plant hormone abscisic acid (ABA) plays a wide range of important roles in plant growth and development, including embryogenesis, seed dormancy, root and shoot growth, transpiration, and stress tolerance. ABA and various abiotic stresses also activate the expression of numerous plant genes through undefined signaling pathways. To gain insight into ABA and stress signal transduction, we conducted a genetic screen based on ABA- and stress-inducible gene transcription. Here we report the identification of an Arabidopsis mutation, fiery1 (fry1), which results in super-induction of ABA- and stress-responsive genes. Seed germination and postembryonic development of fry1 are more sensitive to ABA or stress inhibition. The mutant plants are also compromised in tolerance to freezing, drought, and salt stresses. Map-based cloning revealed that FRY1 encodes an inositol polyphosphate 1-phosphatase, which functions in the catabolism of inositol 1, 4, 5-trisphosphate (IP(3)). Upon ABA treatment, fry1 mutant plants accumulated more IP(3) than did the wild-type plants. These results provide the first genetic evidence indicating that phosphoinositols mediate ABA and stress signal transduction in plants and their turnover is critical for attenuating ABA and stress signaling. PMID- 11485992 TI - BIG: a calossin-like protein required for polar auxin transport in Arabidopsis. AB - Polar auxin transport is crucial for the regulation of auxin action and required for some light-regulated responses during plant development. We have found that two mutants of Arabidopsis-doc1, which displays altered expression of light regulated genes, and tir3, known for its reduced auxin transport-have similar defects and define mutations in a single gene that we have renamed BIG. BIG is very similar to the Drosophila gene Calossin/Pushover, a member of a gene family also present in Caenorhabditis elegans and human genomes. The protein encoded by BIG is extraordinary in size, 560 kD, and contains several putative Zn-finger domains. Expression-profiling experiments indicate that altered expression of multiple light-regulated genes in doc1 mutants can be suppressed by elevated levels of auxin caused by overexpression of an auxin biosynthetic gene, suggesting that normal auxin distribution is required to maintain low-level expression of these genes in the dark. Double mutants of tir3 with the auxin mutants pin1, pid, and axr1 display severe defects in auxin-dependent growth of the inflorescence. Chemical inhibitors of auxin transport change the intracellular localization of the auxin efflux carrier PIN1 in doc1/tir3 mutants, supporting the idea that BIG is required for normal auxin efflux. PMID- 11485993 TI - Distinct mesodermal signals, including BMPs from the septum transversum mesenchyme, are required in combination for hepatogenesis from the endoderm. AB - Mesodermal signaling is critical for patterning the embryonic endoderm into different tissue domains. Classical tissue transplant experiments in the chick and recent studies in the mouse indicated that interactions with the cardiogenic mesoderm are necessary and sufficient to induce the liver in the ventral foregut endoderm. Using molecular markers and functional assays, we now show that septum transversum mesenchyme cells, a distinct mesoderm cell type, are closely apposed to the ventral endoderm and contribute to hepatic induction. Specifically, using a mouse Bmp4 null mutation and an inhibitor of BMPs, we find that BMP signaling from the septum transversum mesenchyme is necessary to induce liver genes in the endoderm and to exclude a pancreatic fate. BMPs apparently function, in part, by affecting the levels of the GATA4 transcription factor, and work in parallel to FGF signaling from the cardiac mesoderm. BMP signaling also appears critical for morphogenetic growth of the hepatic endoderm into a liver bud. Thus, the endodermal domain for the liver is specified by simultaneous signaling from distinct mesodermal sources. PMID- 11485995 TI - DNA topoisomerase VI generates ATP-dependent double-strand breaks with two nucleotide overhangs. AB - A key step in the DNA transport by type II DNA topoisomerase is the formation of a double-strand break with the enzyme being covalently linked to the broken DNA ends (referred to as the cleavage complex). In the present study, we have analyzed the formation and structure of the cleavage complex catalyzed by Sufolobus shibatae DNA topoisomerase VI (topoVI), a member of the recently described type IIB DNA topoisomerase family. A purification procedure of a fully soluble recombinant topoVI was developed by expressing both subunits simultaneously in Escherichia coli. Using this recombinant enzyme, we observed that the formation of the double-strand breaks on supercoiled or linear DNA is strictly dependent on the presence of ATP or AMP-PNP. This result suggests that ATP binding is required to stabilize an enzyme conformation able to cleave the DNA backbone. The structure of cleavage complexes on a linear DNA fragment have been analyzed at the nucleotide level. Similarly to other type II DNA topoisomerases, topoVI is covalently attached to the 5'-ends of the broken DNA. However, sequence analysis of the double-strand breaks revealed that they are all characterized by staggered two-nucleotide long 5' overhangs, contrasting with the four-base staggered double-strand breaks catalyzed by type IIA DNA topoisomerases. While no clear consensus sequences surrounding the cleavage sites could be described, interestingly A and T nucleotides are highly represented on the 5' extensions, giving a first insight on the preferred sequences recognized by this type II DNA topoisomerase. PMID- 11485994 TI - The orphan receptor ALK7 and the Activin receptor ALK4 mediate signaling by Nodal proteins during vertebrate development. AB - Nodal proteins have crucial roles in mesendoderm formation and left-right patterning during vertebrate development. The molecular mechanisms of signal transduction by Nodal and related ligands, however, are not fully understood. In this paper, we present biochemical and functional evidence that the orphan type I serine/threonine kinase receptor ALK7 acts as a receptor for mouse Nodal and Xenopus Nodal-related 1 (Xnr1). Receptor reconstitution experiments indicate that ALK7 collaborates with ActRIIB to confer responsiveness to Xnr1 and Nodal. Both receptors can independently bind Xnr1. In addition, Cripto, an extracellular protein genetically implicated in Nodal signaling, can independently interact with both Xnr1 and ALK7, and its expression greatly enhances the ability of ALK7 and ActRIIB to respond to Nodal ligands. The Activin receptor ALK4 is also able to mediate Nodal signaling but only in the presence of Cripto, with which it can also interact directly. A constitutively activated form of ALK7 mimics the mesendoderm-inducing activity of Xnr1 in Xenopus embryos, whereas a dominant negative ALK7 specifically blocks the activities of Nodal and Xnr1 but has little effect on other related ligands. In contrast, a dominant-negative ALK4 blocks all mesoderm-inducing ligands tested, including Nodal, Xnr1, Xnr2, Xnr4, and Activin. In agreement with a role in Nodal signaling, ALK7 mRNA is localized to the ectodermal and organizer regions of Xenopus gastrula embryos and is expressed during early stages of mouse embryonic development. Therefore, our results indicate that both ALK4 and ALK7 can mediate signal transduction by Nodal proteins, although ALK7 appears to be a receptor more specifically dedicated to Nodal signaling. PMID- 11485996 TI - Identification of a new form of death-associated protein kinase that promotes cell survival. AB - In this study, two alternatively spliced forms of the mouse death-associated protein kinase (DAPK) have been identified and their roles in apoptosis examined. The mouse DAPK-alpha sequence is 95% identical to the previously described human DAPK, and it has a kinase domain and calmodulin-binding region closely related to the 130-150 kDa myosin light chain kinases. A 12-residue extension of the carboxyl terminus of DAPK-beta distinguishes it from the human and mouse DAPK alpha. DAPK phosphorylates at least one substrate in vitro and in vivo, the myosin II regulatory light chain. This phosphorylation occurs preferentially at Ser-19 and is stimulated by calcium and calmodulin. The mRNA encoding DAPK is widely distributed and detected in mouse embryos and most adult tissues, although the expression of the encoded 160-kDa DAPK protein is more restricted. Overexpression of DAPK-alpha, the mouse homolog of human DAPK has a negligible effect on tumor necrosis factor (TNF)-induced apoptosis. Overexpression of DAPK beta has a strong cytoprotective effect on TNF-treated cells. Biochemical analysis of TNF-treated cell lines expressing mouse DAPK-beta suggests that the cytoprotective effect of DAPK is mediated through both intrinsic and extrinsic apoptotic signaling pathways and results in the inhibition of cytochrome c release from the mitochondria as well as inhibition of caspase-3 and caspase-9 activity. These results suggest that the mouse DAPK-beta is a negative regulator of TNF-induced apoptosis. PMID- 11485997 TI - Age-dependent degradation of calpastatin in kidney of hypertensive rats. AB - Hypertensive rats from the Milan strain show a significant decrease in calpastatin activity as compared with normotensive control animals. Calpastatin deficiency is age-related and highly relevant in kidney, heart, and erythrocytes and of minor entity in brain tissue. In normotensives the changes during aging in the levels of calpastatin activity and mRNA are consistent with an increase of calpastatin protein. In hypertensive rats such a relationship during aging is not observed, because a progressive accumulation of mRNA is accompanied by a lower amount of calpastatin protein as compared with control rats. Together with the low level of calpastatin in kidney of hypertensive rats, a progressive accumulation of an active 15-kDa calpastatin fragment, previously shown to represent a typical product of calpain-mediated calpastatin degradation, is also observed. Evidence for such intracellular proteolysis by Ca(2+)-activated calpain is provided by the normalization of the calpastatin level, up to that of control animals, in hypertensive rats treated with drugs known to reduce both blood pressure and intracellular Ca(2+) influx. Further evidence is provided by the disappearance, in these conditions, of the 15-kDa calpastatin fragment. These data allow the conclusion that calpastatin degradation is a relevant part of the overall mechanism for regulating calpain activity. PMID- 11485998 TI - Interactions in the error-prone postreplication repair proteins hREV1, hREV3, and hREV7. AB - Most mutations after DNA damage in yeast Saccharomyces cerevisiae are induced by error-prone translesion DNA synthesis employing scRev1 and DNA polymerase zeta that consists of scRev3 and scRev7 proteins. Recently, the human REV1 (hREV1) and REV3 (hREV3) genes were identified, and their products were revealed to be involved in UV-induced mutagenesis, as observed for their yeast counterparts. Human REV7 (hREV7) was also cloned, and its product was found to interact with hREV3, but the biological function of hREV7 remained unknown. We report here the analyses of precise interactions in the human REV proteins. The interaction between hREV1 and hREV7 was identified by the yeast two-hybrid library screening using a bait of hREV7, which was confirmed by in vitro and in vivo binding assays. The homodimerization of hREV7 was also detected in the two-hybrid analysis. In addition, the precise domains for interaction between hREV7 and hREV1 or hREV3 and for hREV7 homodimerization were determined. Although hREV7 interacts with both hREV1 and hREV3, a stable complex formation of the three proteins was undetectable in vitro. These findings suggest the possibility that hREV7 might play an important role in regulating the enzymatic activities of hREV1 and hREV3 for mutagenesis in response to DNA damage. PMID- 11485999 TI - alpha-Lactalbumin (LA) stimulates milk beta-1,4-galactosyltransferase I (beta 4Gal-T1) to transfer glucose from UDP-glucose to N-acetylglucosamine. Crystal structure of beta 4Gal-T1 x LA complex with UDP-Glc. AB - beta-1,4-Galactosyltransferase 1 (Gal-T1) transfers galactose (Gal) from UDP-Gal to N-acetylglucosamine (GlcNAc), which constitutes its normal galactosyltransferase (Gal-T) activity. In the presence of alpha-lactalbumin (LA), it transfers Gal to Glc, which is its lactose synthase (LS) activity. It also transfers glucose (Glc) from UDP-Glc to GlcNAc, constituting the glucosyltransferase (Glc-T) activity, albeit at an efficiency of only 0.3-0.4% of Gal-T activity. In the present study, we show that LA increases this activity almost 30-fold. It also enhances the Glc-T activity toward various N-acyl substituted glucosamine acceptors. Steady state kinetic studies of Glc-T reaction show that the K(m) for the donor and acceptor substrates are high in the absence of LA. In the presence of LA, the K(m) for the acceptor substrate is reduced 30 fold, whereas for UDP-Glc it is reduced only 5-fold. In order to understand this property, we have determined the crystal structures of the Gal-T1.LA complex with UDP-Glc x Mn(2+) and with N-butanoyl-glucosamine (N-butanoyl-GlcN), a preferred sugar acceptor in the Glc-T activity. The crystal structures reveal that although the binding of UDP-Glc is quite similar to UDP-Gal, there are few significant differences observed in the hydrogen bonding interactions between UDP-Glc and Gal T1. Based on the present kinetic and crystal structural studies, a possible explanation for the role of LA in the Glc-T activity has been proposed. PMID- 11486000 TI - Site specificity of four pyruvate dehydrogenase kinase isoenzymes toward the three phosphorylation sites of human pyruvate dehydrogenase. AB - Activity of the mammalian pyruvate dehydrogenase complex is regulated by phosphorylation-dephosphorylation of three specific serine residues (site 1, Ser 264; site 2, Ser-271; site 3, Ser-203) of the alpha subunit of the pyruvate dehydrogenase (E1) component. Phosphorylation is carried out by four pyruvate dehydrogenase kinase (PDK) isoenzymes. Specificity of the four mammalian PDKs toward the three phosphorylation sites of E1 was investigated using the recombinant E1 mutant proteins with only one functional phosphorylation site present. All four PDKs phosphorylated site 1 and site 2, however, with different rates in phosphate buffer (for site 1, PDK2 > PDK4 approximately PDK1 > PDK3; for site 2, PDK3 > PDK4 > PDK2 > PDK1). Site 3 was phosphorylated by PDK1 only. The maximum activation by dihydrolipoamide acetyltransferase was demonstrated by PDK3. In the free form, all PDKs phosphorylated site 1, and PDK4 had the highest activity toward site 2. The activity of the four PDKs was stimulated to a different extent by the reduction and acetylation state of the lipoyl moieties of dihydrolipoamide acetyltransferase with the maximum stimulation of PDK2. Substitution of the site 1 serine with glutamate, which mimics phosphorylation dependent inactivation of E1, did not affect phosphorylation of site 2 by four PDKs and of site 3 by PDK1. Site specificity for phosphorylation of four PDKs with unique tissue distribution could contribute to the tissue-specific regulation of the pyruvate dehydrogenase complex in normal and pathophysiological states. PMID- 11486001 TI - Identification of inhibitors of TRAIL-induced death (ITIDs) in the TRAIL sensitive colon carcinoma cell line SW480 using a genetic approach. AB - The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent inducer of apoptosis in tumor cell lines, whereas normal cells appear to be protected from its cytotoxic effects. Therefore TRAIL holds promise as a potential therapeutic agent against cancer. To elucidate some of the critical factors that contribute to TRAIL resistance, we performed a genetic screen in the human colon carcinoma cell line SW480 by infecting this TRAIL-sensitive cell line with a human placental cDNA retroviral library and isolating TRAIL-resistant clones. Characterization of the resulting clones for inhibitors of TRAIL-induced death (ITIDs) led to the isolation of c-FLIP(S), Bax inhibitor 1, and Bcl-XL as candidate suppressors of TRAIL signaling. We have demonstrated that c-FLIP(S) and Bcl-XL are sufficient when overexpressed to convey resistance to TRAIL treatment in previously sensitive cell lines. Furthermore both c-FLIP(S) and Bcl-XL protected against overexpression of the TRAIL receptors DR4 and KILLER/DR5. When c-FLIP(S) and Bcl-XL were overexpressed together in SW480 and HCT 116, an additive inhibitory effect was observed after TRAIL treatment suggesting that these two molecules function in the same pathway in the cell lines tested. Furthermore, we have demonstrated for the first time that a proapoptotic member of the Bcl-2 family, Bax, is required for TRAIL-mediated apoptosis in HCT 116 cells. Surprisingly, we have found that the serine/threonine protein kinase Akt, which is an upstream regulator of both c-FLIP(S) and Bcl-XL, is not sufficient when overexpressed to protect against TRAIL in the cell lines tested. These results suggest a key role for c-FLIP(S), Bcl-XL, and Bax in determining tumor cell sensitivity to TRAIL. PMID- 11486002 TI - The structure of human beta-defensin-1: new insights into structural properties of beta-defensins. AB - Defensins are a class of small cationic peptides found in higher organisms that serve as both antimicrobial and cell signaling molecules. The exact mechanism of the antimicrobial activity of defensins is not known, but two models have been postulated, one involving pore formation and the other involving nonspecific electrostatic interaction with the bacterial membrane. Here we report the high resolution structures of human beta-defensin-1 (hBD1) in two crystallographic space groups. The structure of a single molecule is very similar to that of human beta-defensin-2 (hBD2), confirming the presence of an N-terminal alpha-helix. However, while the packing of hBD1 is conserved across both space groups, there is no evidence for any larger quaternary structure similar to octameric hBD2. Furthermore, the topology of hBD1 dimers that are formed between monomers in the asymmetric unit is distinct from both hBD2 and other mammalian alpha-defensins. The structures of hBD1 and hBD2 provide a first step toward understanding the structural basis of antimicrobial and chemotactic properties of human beta defensins. PMID- 11486003 TI - Deletion of specific glycan chains affects differentially the stability, local structures, and activity of lecithin-cholesterol acyltransferase. AB - The enzymatic and interfacial binding activity of lecithin-cholesterol acyltransferase (LCAT) is affected differentially by the location and extent of its glycosylation. Two LCAT glycosylation-deficient mutants, N84Q and N384Q, were constructed, permanently expressed in Chinese hamster ovary cells, and purified to determine the effects of deleting individual glycan chains on its stability, structure, and function. These purified mutants were studied by spectroscopic structural methods and enzymatic and binding assays to develop a molecular rationale for the relationship between LCAT glycosylation and activity. The N84Q LCAT mutant did not possess measurable enzymatic activity or interfacial binding affinity for reconstituted high-density lipoproteins. In addition, in thermal and chemical denaturation studies, N84Q LCAT was found to be significantly less stable than wild-type LCAT. The N384Q variant was initially more enzymatically active than wild-type LCAT, but gradually lost activity within months; however, it retained full interfacial binding activity. Significant changes were detected over time by circular dichroism in the alpha-helical content of N384Q LCAT and in the beta-sheet content of N84Q LCAT, compared with wild-type LCAT. Fluorescence measurements with the probe 1-anilinonapthalene-8-sulfonate suggested an alteration of the active site cavity in both mutants. In conclusion, both mutants lost catalytic activity, N84Q shortly after purification and N384Q more gradually, and were destabilized, probably because the deletion of the glycan chains altered local structural elements near the active site cavity and/or the interfacial binding regions. PMID- 11486004 TI - Remodeling of the major pig xenoantigen by N-acetylglucosaminyltransferase III in transgenic pig. AB - We have been successful in generating several lines of transgenic mice and pigs that contain the human beta-d-mannoside beta-1,4-N-acetylglucosaminyltransferase III (GnT-III) gene. The overexpression of the GnT-III gene in mice and pigs reduced their antigenicity to human natural antibodies, especially the Galalpha1 3Galbeta1-4GlcNAc-R, as evidenced by immunohistochemical analysis. Endothelial cell studies from the GnT-III transgenic pigs also revealed a significant down regulation in antigenicity, including Hanganutziu-Deicher antigen, and dramatic reductions in both the complement- and natural killer cell-mediated pig cell lyses. Changes in the enzymatic activities of other glycosyltransferases, such as alpha1,3-galactosyltransferase, GnT-IV, and GnT-V, did not support cross-talk between GnT-III and these enzymes in the transgenic animals. In addition, we demonstrated the effect of GnT-III in down-regulating the xenoantigen of pig heart grafts, using a pig to cynomolgus monkey transplantation model, suggesting that this approach may be useful in clinical xenotransplantation in the future. PMID- 11486005 TI - Regulation of Snf1 kinase. Activation requires phosphorylation of threonine 210 by an upstream kinase as well as a distinct step mediated by the Snf4 subunit. AB - The yeast Snf1 kinase and its metazoan orthologues, the AMP-activated protein kinases, are activated in response to nutrient limitation. Activation requires the phosphorylation of a conserved threonine residue in the activation loop of the catalytic subunit. A phosphopeptide antibody was generated that specifically recognizes Snf1 protein that is phosphorylated in its activation loop on threonine 210. Using this reagent, we show that phosphorylation of threonine 210 correlates with Snf1 activity, since it is detected in cells subjected to glucose limitation but not in cells grown in abundant glucose. A Snf1 mutant completely lacking kinase activity was phosphorylated normally on threonine 210 in glucose starved cells, eliminating the possibility that the threonine 210 modification is due to an autophosphorylation event. Cells lacking the Reg1 protein, a regulatory subunit for the Glc7 phosphatase, showed constitutive phosphorylation of Snf1 threonine 210. Exposure of cells to high concentrations of sodium chloride also induced phosphorylation of Snf1. Interestingly, Mig1, a downstream target of Snf1 kinase, is phosphorylated in glucose-stressed but not sodium-stressed cells. Finally, cells lacking the gamma subunit of the Snf1 kinase complex encoded by the SNF4 gene exhibited normal regulation of threonine 210 phosphorylation in response to glucose limitation but are unable to phosphorylate Mig1 efficiently. Our data indicate that activation of the Snf1 kinase complex involves two steps, one that requires a distinct upstream kinase and one that is mediated by the gamma subunit of the kinase itself. PMID- 11486007 TI - Differential usage of nuclear export sequences regulates intracellular localization of the dioxin (aryl hydrocarbon) receptor. AB - The dioxin receptor belongs to the basic helix-loop helix/Per-Arnt-Sim (bHLH)/PAS family of proteins and functions as a ligand-dependent transcription factor to activate target genes. The function of the PAS domain of the dioxin receptor is only partially understood. Whereas the C-terminal half of the PAS domain has been shown to harbor ligand binding activity and to function as an accessory dimerization interface, the precise functional role of the N-terminal half of the PAS domain remains unclear. We have previously shown that this domain confers dimerization specificity to the dioxin receptor. Here we report the identification and characterization of a novel nuclear export sequence (NES) motif, located in the N-terminal portion of the PAS domain, in addition to the previously identified NES in the bHLH domain. By point mutagenesis, we have generated a dominant positive form of the PAS domain NES motif that inhibits accumulation of the dioxin receptor in the nuclear compartment of the cell. This mutant form of the receptor was furthermore unable to sustain reporter gene activation. Importantly, we demonstrate that the ligand-free and ligand-occupied forms of the dioxin receptor differentially employ the two NES motifs. In the absence of ligand, nuclear export is sustained via the PAS domain NES, whereas following ligand-dependent activation nuclear export of the receptor is mediated by the NES in the bHLH domain. PMID- 11486006 TI - Synergistic cooperation between hypoxia and transforming growth factor-beta pathways on human vascular endothelial growth factor gene expression. AB - Signaling by transforming growth factor (TGF)-beta family members is mediated by Smad proteins that regulate gene transcription through functional cooperativity and association with other DNA-binding proteins. The hypoxia-inducible factor (HIF)-1 is a transcriptional complex that plays a key role in oxygen-regulated gene expression. We demonstrate that hypoxia and TGF-beta cooperate in the induction of the promoter activity of vascular endothelial growth factor (VEGF), which is a major stimulus in the promotion of angiogenesis. This cooperation has been mapped on the human VEGF promoter within a region at -1006 to -954 that contains functional DNA-binding sequences for HIF-1 and Smads. Optimal HIF-1alpha dependent induction of the VEGF promoter was obtained in the presence of Smad3, suggesting an interaction between these proteins. Consistent with this, co immunoprecipitation experiments revealed that HIF-1alpha physically associates with Smad3. These results demonstrate that both TGF-beta and hypoxia signaling pathways can synergize in the regulation of VEGF gene expression at the transcriptional level. PMID- 11486008 TI - Role of myosin phosphatase isoforms in cGMP-mediated smooth muscle relaxation. AB - In vitro experiments showing the activation of the myosin phosphatase via heterophilic leucine zipper interactions between its targeting subunit (MYPT1) and cGMP-dependent protein kinase I suggested a pathway for smooth muscle relaxation (Surks, H. K., Mochizuki, N., Kasai, Y., Georgescu, S. P., Tang, K. M., Ito, M., Lincoln, T. M., and Mendelsohn, M. E. (1999) Science 286, 1583 1587). The relationship between MYPT1 isoform expression and smooth muscle responses to cGMP signaling in vivo has not been explored. MYPT1 isoforms that contain or lack a C-terminal leucine zipper are generated in birds and mammals by cassette-type alternative splicing of a 31-nucleotide exon. The avian and mammalian C-terminal isoforms are highly conserved and expressed in a tissue specific fashion. In the mature chicken the tonic contracting aorta and phasic contracting gizzard exclusively express the leucine zipper positive and negative MYPT1 isoforms, respectively. Expression of the MYPT1 isoforms is also developmentally regulated in the gizzard, which switches from leucine zipper positive to negative isoforms around the time of hatching. This switch coincides with the development in the gizzard of a cGMP-resistant phenotype, i.e. inability to dephosphorylate myosin and relax in response to 8-bromo-cGMP after calcium activation. Furthermore, association of cGMP-dependent protein kinase I with MYPT1 is detected by immunoprecipitation only in the tissue that expresses the leucine zipper positive isoform of MYPT1. These results suggest that the regulated splicing of MYPT1 is an important determinant of smooth muscle phenotypic diversity and the variability in the response of smooth muscles to the calcium desensitizing effect of cGMP signaling. PMID- 11486009 TI - The high molecular weight urinary matrix metalloproteinase (MMP) activity is a complex of gelatinase B/MMP-9 and neutrophil gelatinase-associated lipocalin (NGAL). Modulation of MMP-9 activity by NGAL. AB - Detection of matrix metalloproteinase (MMP) activities in the urine from patients with a variety of cancers has been closely correlated to disease status. Among these activities, the presence of a group of high molecular weight (HMW) MMPs independently serves as a multivariate predictor of the metastatic phenotype (). The identity of these HMW MMP activities has remained unknown despite their novelty and their potentially important applications in non-invasive cancer diagnosis and/or prognosis. Here, we report the identification of one of these HMW urinary MMPs of approximately 125-kDa as being a complex of gelatinase B (MMP 9) and neutrophil gelatinase-associated lipocalin (NGAL). Multiple biochemical approaches verified this identity. Analysis using substrate gel electrophoresis demonstrated that the 125-kDa urinary MMP activity co-migrates with purified human neutrophil MMP-9 x NGAL complex. The 125-kDa urinary MMP-9 x NGAL complex was recognized by a purified antibody against human NGAL as well as by a monospecific anti-human MMP-9 antibody. Furthermore, these same two antibodies were independently capable of specifically immunoprecipitating the 125-kDa urinary MMP activity in a dose-dependent manner. In addition, the complex of MMP 9 x NGAL could be reconstituted in vitro by mixing MMP-9 and NGAL in gelatinase buffers with pH values in the range of urine and in normal urine as well. Finally, the biochemical consequences of the NGAL and MMP-9 interaction were investigated both in vitro using recombinant human NGAL and MMP-9 and in cell culture by overexpressing NGAL in human breast carcinoma cells. Our data demonstrate that NGAL is capable of protecting MMP-9 from degradation in a dose dependent manner and thereby preserving MMP-9 enzymatic activity. In summary, this study identifies the 125-kDa urinary gelatinase as being a complex of MMP-9 and NGAL and provides evidence that NGAL modulates MMP-9 activity by protecting it from degradation. PMID- 11486010 TI - Expression of the three human major histocompatibility complex class II isotypes exhibits a differential dependence on the transcription factor RFXAP. AB - Major histocompatibility complex class II (MHCII) molecules play a pivotal role in the immune system because they direct the development and activation of CD4(+) T cells. There are three human MHCII isotypes, HLA-DR, HLA-DQ, and HLA-DP. Key transcription factors controlling MHCII genes have been identified by virtue of the fact that they are mutated in a hereditary immunodeficiency resulting from a lack of MHCII expression. RFXAP-one of the factors affected in this disease-is a subunit of RFX, a DNA-binding complex that recognizes the X box present in all MHCII promoters. To facilitate identification of conserved regions in RFXAP, we isolated the mouse gene. We then delimited conserved domains required to restore endogenous MHCII expression in cell lines lacking a functional RFXAP gene. Surprisingly, we found that 80% of RFXAP is dispensable for the reactivation of DR expression. Only a short C-terminal segment of the protein is essential for this isotype. In contrast, optimal expression of DQ and DP requires a larger C terminal segment. These results define an RFXAP domain with an MHCII isotype specific function. Expression of the three MHCII isotypes exhibits a differential requirement for this domain. We show that this is due to a differential dependence on this domain for promoter occupation and recruitment of the coactivator CIITA in vivo. PMID- 11486011 TI - Inhibition of acetyl coenzyme A carboxylase activity restores expression of the INO1 gene in a snf1 mutant strain of Saccharomyces cerevisiae. AB - Mutations in the Saccharomyces cerevisiae SNF1 gene affect a number of cellular processes, including the expression of genes involved in carbon source utilization and phospholipid biosynthesis. To identify targets of the Snf1 kinase that modulate expression of INO1, a gene required for an early, rate-limiting step in phospholipid biosynthesis, we performed a genetic selection for suppressors of the inositol auxotrophy of snf1Delta strains. We identified mutations in ACC1 and FAS1, two genes important for fatty acid biosynthesis in yeast; ACC1 encodes acetyl coenzyme A carboxylase (Acc1), and FAS1 encodes the beta subunit of fatty acid synthase. Acc1 was shown previously to be phosphorylated and inactivated by Snf1. Here we show that snf1Delta strains with increased Acc1 activity exhibit decreased INO1 transcription. Strains carrying the ACC1 suppressor mutation have reduced Acc1 activity in vitro and in vivo, as revealed by enzymatic assays and increased sensitivity to the Acc1-specific inhibitor soraphen A. Moreover, a reduction in Acc1 activity, caused by addition of soraphen A, provision of exogenous fatty acid, or conditional expression of ACC1, suppresses the inositol auxotrophy of snf1Delta strains. Together, these findings indicate that the inositol auxotrophy of snf1Delta strains arises in part from elevated Acc1 activity and that a reduction in this activity restores INO1 expression in these strains. These results reveal a Snf1-dependent connection between fatty acid production and phospholipid biosynthesis, identify Acc1 as a Snf1 target important for INO1 transcription, and suggest models in which metabolites that are generated or utilized during fatty acid biosynthesis can significantly influence gene expression in yeast. PMID- 11486012 TI - HMG box transcriptional repressor HBP1 maintains a proliferation barrier in differentiated liver tissue. AB - We previously isolated HBP1 as a target of the retinoblastoma (RB) and p130 family members and as the first of the HMG box transcriptional repressors. Our subsequent work demonstrated that HBP1 coordinates differentiation in cell culture models. In the present study, we show that HBP1 regulates proliferation in a differentiated tissue of an animal. Using transgenic mice in which HBP1 expression was specifically increased in hepatocytes under control of the transthyretin promoter, we determined the impact of HBP1 on synchronous cell cycle reentry following partial hepatectomy. Modest overexpression of HBP1 yielded a detectable cell cycle phenotype. Following a mitogenic stimulus induced by two-thirds partial hepatectomy, mice expressing the HBP1 transgene showed a 10 to 12-h delay in progression through G(1) to the peak of S phase. There was a concomitant delay in mid-G(1) events, such as the induction of cyclin E. While the delay in G(1) and S phases correlated with the slight overexpression of transgenic HBP1, the level of the endogenous HBP1 protein itself declined in S phase. In contrast, the onset of the immediate-early response following partial hepatectomy was unchanged in HBP1 transgenic mice. This observation indicated that the observed delay in S phase did not result from changes in signaling pathways leading into the G(0)-to-G(1) transition. Finally, transgenic mice expressing a mutant HBP1 lacking the N-terminal RB interacting domain showed a stronger S-phase response following partial hepatectomy. These results provide the first evidence that HBP1 can regulate cell cycle progression in differentiated tissues. PMID- 11486013 TI - Evidence that fungal MEP proteins mediate diffusion of the uncharged species NH(3) across the cytoplasmic membrane. AB - Methylammonium and ammonium (MEP) permeases of Saccharomyces cerevisiae belong to a ubiquitous family of cytoplasmic membrane proteins that transport only ammonium (NH(4)(+) + NH(3)). Transport and accumulation of the ammonium analog [(14)C]methylammonium, a weak base, led to the proposal that members of this family were capable of energy-dependent concentration of the ammonium ion, NH(4)(+). In bacteria, however, ATP-dependent conversion of methylammonium to gamma-N-methylglutamine by glutamine synthetase precludes its use in assessing concentrative transport across the cytoplasmic membrane. We have confirmed that methylammonium is not metabolized in the yeast S. cerevisiae and have shown that it is little metabolized in the filamentous fungus Neurospora crassa. However, its accumulation depends on the energy-dependent acidification of vacuoles. A Deltavph1 mutant of S. cerevisiae and a Deltavma1 mutant, which lack vacuolar H(+)-ATPase activity, had large (fivefold or greater) defects in the accumulation of methylammonium, with little accompanying defect in the initial rate of transport. A vma-1 mutant of N. crassa largely metabolized methylammonium to methylglutamine. Thus, in fungi as in bacteria, subsequent energy-dependent utilization of methylammonium precludes its use in assessing active transport across the cytoplasmic membrane. The requirement for a proton gradient to sequester the charged species CH(3)NH(3)(+) in acidic vacuoles provides evidence that the substrate for MEP proteins is the uncharged species CH(3)NH(2). By inference, their natural substrate is NH(3), a gas. We postulate that MEP proteins facilitate diffusion of NH(3) across the cytoplasmic membrane and speculate that human Rhesus proteins, which lie in the same domain family as MEP proteins, facilitate diffusion of CO(2). PMID- 11486014 TI - Antagonistic controls of autophagy and glycogen accumulation by Snf1p, the yeast homolog of AMP-activated protein kinase, and the cyclin-dependent kinase Pho85p. AB - In the yeast Saccharomyces cerevisiae, glycogen is accumulated as a carbohydrate reserve when cells are deprived of nutrients. Yeast mutated in SNF1, a gene encoding a protein kinase required for glucose derepression, has diminished glycogen accumulation and concomitant inactivation of glycogen synthase. Restoration of synthesis in an snf1 strain results only in transient glycogen accumulation, implying the existence of other SNF1-dependent controls of glycogen storage. A genetic screen revealed that two genes involved in autophagy, APG1 and APG13, may be regulated by SNF1. Increased autophagic activity was observed in wild-type cells entering the stationary phase, but this induction was impaired in an snf1 strain. Mutants defective for autophagy were able to synthesize glycogen upon approaching the stationary phase, but were unable to maintain their glycogen stores, because subsequent synthesis was impaired and degradation by phosphorylase, Gph1p, was enhanced. Thus, deletion of GPH1 partially reversed the loss of glycogen accumulation in autophagy mutants. Loss of the vacuolar glucosidase, SGA1, also protected glycogen stores, but only very late in the stationary phase. Gph1p and Sga1p may therefore degrade physically distinct pools of glycogen. Pho85p is a cyclin-dependent protein kinase that antagonizes SNF1 control of glycogen synthesis. Induction of autophagy in pho85 mutants entering the stationary phase was exaggerated compared to the level in wild-type cells, but was blocked in apg1 pho85 mutants. We propose that Snf1p and Pho85p are, respectively, positive and negative regulators of autophagy, probably via Apg1 and/or Apg13. Defective glycogen storage in snf1 cells can be attributed to both defective synthesis upon entry into stationary phase and impaired maintenance of glycogen levels caused by the lack of autophagy. PMID- 11486015 TI - In vitro reconstitution of the end replication problem. AB - The end replication problem hypothesis proposes that the ends of linear DNA cannot be replicated completely during lagging strand DNA synthesis. Although the idea has been widely accepted for explaining telomere attrition during cell proliferation, it has never been directly demonstrated. In order to take a biochemical approach to understand how linear DNA ends are replicated, we have established a novel in vitro linear simian virus 40 DNA replication system. In this system, terminally biotin-labeled linear DNAs are conjugated to avidin coated beads and subjected to replication reactions. Linear DNA was efficiently replicated under optimized conditions, and replication products that had replicated using the original DNA templates were specifically analyzed by purifying bead-bound replication products. By exploiting this system, we showed that while the leading strand is completely synthesized to the end, lagging strand synthesis is gradually halted in the terminal approximately 500-bp region, leaving 3' overhangs. This result is consistent with observations in telomerase negative mammalian cells and formally demonstrates the end replication problem. This study provides a basis for studying the details of telomere replication. PMID- 11486016 TI - Control of DNA rereplication via Cdc2 phosphorylation sites in the origin recognition complex. AB - Cdc2 kinase is a master regulator of cell cycle progression in the fission yeast Schizosaccharomyces pombe. Our data indicate that Cdc2 phosphorylates replication factor Orp2, a subunit of the origin recognition complex (ORC). Cdc2 phosphorylation of Orp2 appears to be one of multiple mechanisms by which Cdc2 prevents DNA rereplication in a single cell cycle. Cdc2 phosphorylation of Orp2 is not required for Cdc2 to activate DNA replication initiation. Phosphorylation of Orp2 appears first in S phase and becomes maximal in G(2) and M when Cdc2 kinase activity is required to prevent reinitiation of DNA replication. A mutant lacking Cdc2 phosphorylation sites in Orp2 (orp2-T4A) allowed greater rereplication of DNA than congenic orp2 wild-type strains when the limiting replication initiation factor Cdc18 was deregulated. Thus, Cdc2 phosphorylation of Orp2 may be redundant with regulation of Cdc18 for preventing reinitiation of DNA synthesis. Since Cdc2 phosphorylation sites are present in Orp2 (also known as Orc2) from yeasts to metazoans, we propose that cell cycle-regulated phosphorylation of the ORC provides a safety net to prevent DNA rereplication and resulting genetic instability. PMID- 11486017 TI - Parallel and independent regulation of interleukin-3 mRNA turnover by phosphatidylinositol 3-kinase and p38 mitogen-activated protein kinase. AB - AU-rich elements (ARE) present in the 3' untranslated regions of many cytokines and immediate-early genes are responsible for targeting the transcripts for rapid decay. We present evidence from cotransfection experiments in NIH 3T3 cells that two signaling pathways, one involving phosphatidylinositol 3-kinase (PI3-K), and one involving the p38 mitogen-activated protein kinase (MAPK), lead to stabilization of interleukin-3 mRNA in parallel. Stabilization mediated by either of the two pathways was antagonized by tristetraprolin (TTP), an AU-binding protein known to promote constitutive decay of ARE-containing transcripts. Remarkably, the stabilizing AU-binding protein HuR, in collaboration with p38 MAPK but not with PI3-K, could overcome the destabilizing effect of TTP. These data argue that the stabilizing kinases PI3-K and p38 MAPK do not act through direct inactivation of TTP but via activating pathway-specific stabilizing AU binding proteins. Our data suggest an integrated model of mRNA turnover control, where stabilizing (HuR) and destabilizing (TTP) AU-binding proteins compete and where the former are under the positive control of independent phosphokinase signaling pathways. PMID- 11486018 TI - Interaction of the Srb10 kinase with Sip4, a transcriptional activator of gluconeogenic genes in Saccharomyces cerevisiae. AB - Sip4 is a Zn(2)Cys(6) transcriptional activator that binds to the carbon source responsive elements of gluconeogenic genes in Saccharomyces cerevisiae. The Snf1 protein kinase interacts with Sip4 and regulates its phosphorylation and activator function in response to glucose limitation; however, evidence suggested that another kinase also regulates Sip4. Here we examine the role of the Srb10 kinase, a component of the RNA polymerase II holoenzyme that has been primarily implicated in transcriptional repression but also positively regulates Gal4. We show that Srb10 is required for phosphorylation of Sip4 during growth in nonfermentable carbon sources and that the catalytic activity of Srb10 stimulates the ability of LexA-Sip4 to activate transcription of a reporter. Srb10 and Sip4 coimmunoprecipitate from cell extracts and interact in two-hybrid assays, suggesting that Srb10 regulates Sip4 directly. We also present evidence that the Srb10 and Snf1 kinases interact with different regions of Sip4. These findings support the view that the Srb10 kinase not only plays negative roles in transcriptional control but also has broad positive roles during growth in carbon sources other than glucose. PMID- 11486019 TI - c-Myb transcription is activated by protein kinase B (PKB) following interleukin 2 stimulation of Tcells and is required for PKB-mediated protection from apoptosis. AB - During T-cell activation, c-Myb is induced upon interleukin 2 (IL-2) stimulation and is required for correct proliferation of cells. In this paper, we provide evidence that IL-2-mediated induction of the c-myb gene occurs via the phosphoinositide 3-kinase (PI3K) signaling pathway, that protein kinase B (PKB) is the principal transducer of this signal, and that activation of the c-myb promoter can be abolished by deletion of conserved E2F and NF-kappaB binding sites. We show that Myb is required to protect activated peripheral T cells from bcl-2-independent apoptosis and that overexpression of oncogenic v-Myb is antiapoptotic. Overexpression of a Myb dominant-negative transgene abrogates PKB mediated protection from apoptosis. Taken together, these results suggest that induction of c-myb transcription is an important downstream event for PKB mediated protection of T cells from programmed cell death. PMID- 11486020 TI - Overlapping functions of the pRb family in the regulation of rRNA synthesis. AB - The "pocket" proteins pRb, p107, and p130 are a family of negative growth regulators. Previous studies have demonstrated that overexpression of pRb can repress transcription by RNA polymerase (Pol) I. To assess whether pRb performs this role under physiological conditions, we have examined pre-rRNA levels in cells from mice lacking either pRb alone or combinations of the three pocket proteins. Pol I transcription was unaffected in pRb-knockout fibroblasts, but specific disruption of the entire pRb family deregulated rRNA synthesis. Further analysis showed that p130 shares with pRb the ability to repress Pol I transcription, whereas p107 is ineffective in this system. Production of rRNA is abnormally elevated in Rb(-/-) p130(-/-) fibroblasts. Furthermore, overexpression of p130 can inhibit an rRNA promoter both in vitro and in vivo. This reflects an ability of p130 to bind and inactivate the upstream binding factor, UBF. The data imply that rRNA synthesis in living cells is subject to redundant control by endogenous pRb and p130. PMID- 11486021 TI - Promoter clearance by RNA polymerase II is an extended, multistep process strongly affected by sequence. AB - We have characterized RNA polymerase II complexes halted from +16 to +49 on two templates which differ in the initial 20 nucleotides (nt) of the transcribed region. On a template with a purine-rich initial transcript, most complexes halted between +20 and +32 become arrested and cannot resume RNA synthesis without the SII elongation factor. These arrested complexes all translocate upstream to the same location, such that about 12 to 13 bases of RNA remain in each of the complexes after SII-mediated transcript cleavage. Much less arrest is observed over this same region with a second template in which the initially transcribed region is pyrimidine rich, but those complexes which do arrest on the second template also translocate upstream to the same location observed with the first template. Complexes stalled at +16 to +18 on either template do not become arrested. Complexes stalled at several locations downstream of +35 become partially arrested, but these more promoter-distal arrested complexes translocate upstream by less than 10 nt; that is, they do not translocate to a common, far upstream location. Kinetic studies with nonlimiting levels of nucleoside triphosphates reveal strong pausing between +20 and +30 on both templates. These results indicate that promoter clearance by RNA polymerase II is at least a two step process: a preclearance escape phase extending up to about +18 followed by an unstable clearance phase which extends over the formation of 9 to 17 more bonds. Polymerases halted during the clearance phase translocate upstream to the preclearance location and arrest in at least one sequence context. PMID- 11486022 TI - Transcriptional activation domains of human heat shock factor 1 recruit human SWI/SNF. AB - Chromatin remodeling complexes such as SWI/SNF use the energy of ATP hydrolysis to remodel nucleosomal DNA and increase transcription of nucleosomal templates. Human heat shock factor one (hHSF1) is a tightly regulated activator that stimulates transcriptional initiation and elongation using different portions of its activation domains. Here we demonstrate that hHSF1 associates with BRG1, the ATPase subunit of human SWI/SNF (hSWI/SNF) at endogenous protein concentrations. We also show that hHSF1 activation domains recruit hSWI/SNF to a chromatin template in a purified system. Mutation of hHSF1 residues responsible for activation of transcriptional elongation has the most severe effect on recruitment of SWI/SNF and association of hHSF1 with BRG1, suggesting that recruitment of chromatin remodeling activity might play a role in stimulation of elongation. PMID- 11486023 TI - Chl12 (Ctf18) forms a novel replication factor C-related complex and functions redundantly with Rad24 in the DNA replication checkpoint pathway. AB - RAD24 has been identified as a gene essential for the DNA damage checkpoint in budding yeast. Rad24 is structurally related to subunits of the replication factor C (RFC) complex, and forms an RFC-related complex with Rfc2, Rfc3, Rfc4, and Rfc5. The rad24Delta mutation enhances the defect of rfc5-1 in the DNA replication block checkpoint, implicating RAD24 in this checkpoint. CHL12 (also called CTF18) encodes a protein that is structurally related to the Rad24 and RFC proteins. We show here that although neither chl12Delta nor rad24Delta single mutants are defective, chl12Delta rad24Delta double mutants become defective in the replication block checkpoint. We also show that Chl12 interacts physically with Rfc2, Rfc3, Rfc4, and Rfc5 and forms an RFC-related complex which is distinct from the RFC and RAD24 complexes. Our results suggest that Chl12 forms a novel RFC-related complex and functions redundantly with Rad24 in the DNA replication block checkpoint. PMID- 11486024 TI - Role of phosphoinositide 3-kinase in the aggressive tumor growth of HT1080 human fibrosarcoma cells. AB - We have developed a model system of human fibrosarcoma cell lines that do or do not possess and express an oncogenic mutant allele of N-ras. HT1080 cells contain an endogenous mutant allele of N-ras, whereas the derivative MCH603 cell line contains only wild-type N-ras. In an earlier study (S. Gupta et al., Mol. Cell. Biol. 20:9294-9306, 2000), we had shown that HT1080 cells produce rapidly growing, aggressive tumors in athymic nude mice, whereas MCH603 cells produced more slowly growing tumors and was termed weakly tumorigenic. An extensive analysis of the Ras signaling pathways (Raf, Rac1, and RhoA) provided evidence for a potential novel pathway that was critical for the aggressive tumorigenic phenotype and could be activated by elevated levels of constitutively active MEK. In this study we examined the role of phosphoinositide 3-kinase (PI 3-kinase) in the regulation of the transformed and aggressive tumorigenic phenotypes expressed in HT1080 cells. Both HT1080 (mutant N-ras) and MCH603 (wild-type N-ras) have similar levels of constitutively active Akt, a downstream target of activated PI 3-kinase. We find that both cell lines constitutively express platelet-derived growth factor (PDGF) and PDGF receptors. Transfection with tumor suppressor PTEN cDNA into HT1080 and constitutively active PI 3-kinase-CAAX cDNA into MCH603 cells, respectively, resulted in several interesting and novel observations. Activation of the PI 3-kinase/Akt pathway, including NF-kappaB, is not required for the aggressive tumorigenic phenotype in HT1080 cells. Activation of NF-kappaB is complex: in MCH603 cells it is mediated by Akt, whereas in HT1080 cells activation also involves other pathway(s) that are activated by mutant Ras. A threshold level of activation of PI 3-kinase is required in MCH603 cells before stimulatory cross talk to the RhoA, Rac1, and Raf pathways occurs, without a corresponding activation of Ras. The increased levels of activation seen were similar to those observed in HT1080 cells, except for Raf and MEK, which were more active than HT1080 levels. This cross talk results in conversion to the aggressive tumorigenic phenotype. This latter observation is consistent with our previous observation that overstimulation of the activity of endogenous members of Ras signaling pathways, activated MEK in particular, is a prerequisite for aggressive tumorigenic growth. PMID- 11486025 TI - Oncogenic mutants of RON and MET receptor tyrosine kinases cause activation of the beta-catenin pathway. AB - beta-Catenin is an oncogenic protein involved in regulation of cell-cell adhesion and gene expression. Accumulation of cellular beta-catenin occurs in many types of human cancers. Four mechanisms are known to cause increases in beta-catenin: mutations of beta-catenin, adenomatous polyposis coli, or axin genes and activation of Wnt signaling. We report a new cause of beta-catenin accumulation involving oncogenic mutants of RON and MET receptor tyrosine kinases (RTKs). Cells transfected with oncogenic RON or MET were characterized by beta-catenin tyrosine phosphorylation and accumulation; constitutive activation of a Tcf transcriptional factor; and increased levels of beta-catenin/Tcf target oncogene proteins c-myc and cyclin D1. Interference with the beta-catenin pathway reduced the transforming potential of mutated RON and MET. Activation of beta-catenin by oncogenic RON and MET constitutes a new pathway, which might lead to cell transformation by these and other mutant growth factor RTKs. PMID- 11486026 TI - c-Abl regulates p53 levels under normal and stress conditions by preventing its nuclear export and ubiquitination. AB - The p53 protein is subject to Mdm2-mediated degradation by the ubiquitin proteasome pathway. This degradation requires interaction between p53 and Mdm2 and the subsequent ubiquitination and nuclear export of p53. Exposure of cells to DNA damage results in the stabilization of the p53 protein in the nucleus. However, the underlying mechanism of this effect is poorly defined. Here we demonstrate a key role for c-Abl in the nuclear accumulation of endogenous p53 in cells exposed to DNA damage. This effect of c-Abl is achieved by preventing the ubiquitination and nuclear export of p53 by Mdm2, or by human papillomavirus E6. c-Abl null cells fail to accumulate p53 efficiently following DNA damage. Reconstitution of these cells with physiological levels of c-Abl is sufficient to promote the normal response of p53 to DNA damage via nuclear retention. Our results help to explain how p53 is accumulated in the nucleus in response to DNA damage. PMID- 11486027 TI - Structural and functional analysis of an mRNP complex that mediates the high stability of human beta-globin mRNA. AB - Human globins are encoded by mRNAs exhibiting high stabilities in transcriptionally silenced erythrocyte progenitors. Unlike alpha-globin mRNA, whose stability is enhanced by assembly of a specific messenger RNP (mRNP) alpha complex on its 3' untranslated region (UTR), neither the structure(s) nor the mechanism(s) that effects the high-level stability of human beta-globin mRNA has been identified. The present work describes an mRNP complex assembling on the 3' UTR of the beta-globin mRNA that exhibits many of the properties of the stability enhancing alpha complex. The beta-globin mRNP complex is shown to contain one or more factors homologous to alphaCP, a 39-kDa RNA-binding protein that is integral to alpha-complex assembly. Sequence analysis implicates a specific 14-nucleotide pyrimidine-rich track within its 3' UTR as the site of beta-globin mRNP assembly. The importance of this track to mRNA stability is subsequently verified in vivo using mice expressing human beta-globin transgenes that contain informative mutations in this region. In combination, the in vitro and in vivo analyses indicate that the high stabilities of the alpha- and beta-globin mRNAs are maintained through related mRNP complexes that may share a common regulatory pathway. PMID- 11486028 TI - Loss of HuR is linked to reduced expression of proliferative genes during replicative senescence. AB - Cellular aging is accompanied by alterations in gene expression patterns. Here, using two models of replicative senescence, we describe the influence of the RNA binding protein HuR in regulating the expression of several genes whose expression decreases during senescence. We demonstrate that HuR levels, HuR binding to target mRNAs encoding proliferative genes, and the half-lives of such mRNAs are lower in senescent cells. Importantly, overexpression of HuR in senescent cells restored a "younger" phenotype, while a reduction in HuR expression accentuated the senescent phenotype. Our studies highlight a critical role for HuR during the process of replicative senescence. PMID- 11486029 TI - Growth factors can influence cell growth and survival through effects on glucose metabolism. AB - Cells from multicellular organisms are dependent upon exogenous signals for survival, growth, and proliferation. The relationship among these three processes was examined using an interleukin-3 (IL-3)-dependent cell line. No fixed dose of IL-3 determined the threshold below which cells underwent apoptosis. Instead, increasing growth factor concentrations resulted in progressive shortening of the G(1) phase of the cell cycle and more rapid proliferative expansion. Increased growth factor concentrations also resulted in proportional increases in glycolytic rates. Paradoxically, cells growing in high concentrations of growth factor had an increased susceptibility to cell death upon growth factor withdrawal. This susceptibility correlated with the magnitude of the change in the glycolytic rate following growth factor withdrawal. To investigate whether changes in the availability of glycolytic products influence mitochondrion initiated apoptosis, we artificially limited glycolysis by manipulating the glucose levels in the medium. Like growth factor withdrawal, glucose limitation resulted in Bax translocation, a decrease in mitochondrial membrane potential, and cytochrome c redistribution to the cytosol. In contrast, increasing cell autonomous glucose uptake by overexpression of Glut1 significantly delayed apoptosis following growth factor withdrawal. These data suggest that a primary function of growth factors is to regulate glucose uptake and metabolism and thus maintain mitochondrial homeostasis and enable anabolic pathways required for cell growth. Consistent with this hypothesis, expression of the three genes involved in glucose uptake and glycolytic commitment, those for Glut1, hexokinase 2, and phosphofructokinase 1, was found to rapidly decline to nearly undetectable levels following growth factor withdrawal. PMID- 11486030 TI - AMF1 (GPS2) modulates p53 transactivation. AB - We have reported that the papillomavirus E2 protein binds the nuclear factor AMF1 (also called G-protein pathway suppressor 2 or GPS2) and that their interaction is necessary for transcriptional activation by E2. It has also been shown that AMF1 can influence the activity of cellular transcription factors. These observations led us to test whether AMF1 regulates the functions of p53, a critical transcriptional activator that integrates stress signals and regulates cell cycle and programmed cell death. We report that AMF1 associates with p53 in vivo and in vitro and facilitates the p53 response by augmenting p53-dependent transcription. Overexpression of AMF1 in U2OS cells increases basal level p21(WAF1/CIP1) expression and causes a G(1) arrest. U2OS cells stably overexpressing AMF1 show increased apoptosis upon exposure to UV irradiation. These data demonstrate that AMF1 modulates p53 activities. PMID- 11486031 TI - Induction of cyclin D1 transcription and CDK2 activity by Notch(ic): implication for cell cycle disruption in transformation by Notch(ic). AB - Notch genes encode a family of transmembrane proteins that are involved in many cellular processes such as differentiation, proliferation, and apoptosis. Although it is well established that all four Notch genes can act as oncogenes, the mechanism by which Notch proteins transform cells remains unknown. Previously, we have shown that transformation of RKE cells can be conditionally induced by hormone activation of Notch(ic)-estrogen receptor (ER) chimeras. Using this inducible system, we show that Notch(ic) activates transcription of the cyclin D1 gene with rapid kinetics. Transcriptional activation of cyclin D1 is independent from serum-derived growth factors and de novo synthesis of secondary transcriptional activators. Moreover, hormone activation of Notch(ic)-ER proteins induces CDK2 activity in the absence of serum. Upregulation of cyclin D1 and activation of CDK2 by Notch(ic) result in the promotion of S-phase entry. These data demonstrate the first evidence that Notch(ic) proteins can directly regulate factors involved in cell cycle control and affect cellular proliferation. Furthermore, nontransforming Notch(ic) proteins do not induce cyclin D1 expression, indicating that the mechanism of transformation involves cell cycle deregulation through constitutive expression of cyclin D1. Finally, we have identified a CSL [stands for CBF1, Su(H), and Lag-1] binding site within the human and rat cyclin D1 promoters, suggesting that Notch(ic) proteins activate cyclin D1 transcription through a CSL-dependent pathway. PMID- 11486032 TI - The E2A-HLF oncoprotein activates Groucho-related genes and suppresses Runx1. AB - The E2A-HLF fusion gene, formed by the t(17;19)(q22;p13) chromosomal translocation in leukemic pro-B cells, encodes a chimeric transcription factor consisting of the transactivation domain of E2A linked to the bZIP DNA-binding and protein dimerization domain of hepatic leukemia factor (HLF). This oncoprotein blocks apoptosis induced by growth factor deprivation or irradiation, but the mechanism for this effect remains unclear. We therefore performed representational difference analysis (RDA) to identify downstream genetic targets of E2A-HLF, using a murine FL5.12 pro-B cell line that had been stably transfected with E2A-HLF cDNA under the control of a zinc-regulated metallothionein promoter. Two RDA clones, designated RDA1 and RDA3, were differentially upregulated in E2A-HLF-positive cells after zinc induction. The corresponding cDNAs encoded two WD40 repeat-containing proteins, Grg2 and Grg6. Both are related to the Drosophila protein Groucho, a transcriptional corepressor that lacks DNA-binding activity on its own but can act in concert with other proteins to regulate embryologic development of the fly. Expression of both Grg2 and Grg6 was upregulated 10- to 50-fold by E2A-HLF. Immunoblot analysis detected increased amounts of two additional Groucho-related proteins, Grg1 and Grg4, in cells expressing E2A-HLF. A mutant E2A-HLF protein with a disabled DNA-binding region also mediated pro-B cell survival and activated Groucho-related genes. Among the transcription factors known to interact with Groucho-related protein, only RUNX1 was appreciably downregulated by E2A-HLF. Our results identify a highly conserved family of transcriptional corepressors that are activated by E2A HLF, and they suggest that downregulation of RUNX1 may contribute to E2A-HLF mediated leukemogenesis. PMID- 11486033 TI - Identification of receptor and heparin binding sites in fibroblast growth factor 4 by structure-based mutagenesis. AB - Fibroblast growth factors (FGFs) comprise a large family of multifunctional, heparin-binding polypeptides that show diverse patterns of interaction with a family of receptors (FGFR1 to -4) that are subject to alternative splicing. FGFR binding specificity is an essential mechanism in the regulation of FGF signaling and is achieved through primary sequence differences among FGFs and FGFRs and through usage of two alternative exons, IIIc and IIIb, for the second half of immunoglobulin-like domain 3 (D3) in FGFRs. While FGF4 binds and activates the IIIc splice forms of FGFR1 to -3 at comparable levels, it shows little activity towards the IIIb splice forms of FGFR1 to -3 as well as towards FGFR4. To begin to explore the structural determinants for this differential affinity, we determined the crystal structure of FGF4 at a 1.8-A resolution. FGF4 adopts a beta-trefoil fold similar to other FGFs. To identify potential receptor and heparin binding sites in FGF4, a ternary FGF4-FGFR1-heparin model was constructed by superimposing the FGF4 structure onto FGF2 in the FGF2-FGFR1-heparin structure. Mutation of several key residues in FGF4, observed to interact with FGFR1 or with heparin in the model, produced ligands with reduced receptor binding and concomitant low mitogenic potential. Based on the modeling and mutational data, we propose that FGF4, like FGF2, but unlike FGF1, engages the betaC'-betaE loop in D3 and thus can differentiate between the IIIc and IIIb splice isoforms of FGFRs for binding. Moreover, we show that FGF4 needs to interact with both the 2-O- and 6-O-sulfates in heparin to exert its optimal biological activity. PMID- 11486034 TI - Signal pathways which promote invasion and metastasis: critical and distinct contributions of extracellular signal-regulated kinase and Ral-specific guanine exchange factor pathways. AB - Approximately 50% of metastatic tumors contain Ras mutations. Ras proteins can activate at least three downstream signaling cascades mediated by the Raf-MEK extracellular signal-regulated kinase family, phosphatidylinositol-3 (PI3) kinase, and Ral-specific guanine nucleotide exchange factors (RalGEFs). Here we investigated the contribution of RalGEF and ERK activation to the development of experimental metastasis in vivo and associated invasive properties in vitro. Each pathway contributes distinct properties to the metastatic phenotype. Following lateral tail vein injection, 3T3 cells transformed by constitutively active Raf or MEK produced lung metastasis that displayed circumscribed, noninfiltrating borders. In contrast, 3T3 cells transformed by Ras(12V,37G), a Ras effector mutant that activates RalGEF but not Raf or P13 kinase, formed aggressive, infiltrative metastasis. Dominant negative RalB inhibited Ras(12V,37G)-activated invasion and metastasis, demonstrating the necessity of the RalGEF pathway for a fully transformed phenotype. Moreover, 3T3 cells constitutively expressing a membrane-associated form of RalGEF (RalGDS-CAAX) formed invasive tumors as well, demonstrating that activation of a RalGEF pathway is sufficient to initiate the invasive phenotype. Despite the fact that Ras(12V,37G) expression does not elevate ERK activity, inhibition of this kinase by a conditionally expressed ERK phosphatase demonstrated that ERK activity was necessary for Ras(12V,37G) transformed cells to express matrix-degrading activity in vitro and tissue invasiveness in vivo. Therefore, these experiments have revealed a hitherto unknown but essential interaction of the RalGEF and ERK pathways to produce a malignant phenotype. The generality of the role of the RalGEF pathway in metastasis is supported by the finding that Ras(12V,37G) increased the invasiveness of epithelial cells as well as fibroblasts. PMID- 11486035 TI - Neurocan is dispensable for brain development. AB - Neurocan is a component of the extracellular matrix in brain. Due to its inhibition of neuronal adhesion and outgrowth in vitro and its expression pattern in vivo it was suggested to play an important role in axon guidance and neurite growth. To study the role of neurocan in brain development we generated neurocan deficient mice by targeted disruption of the neurocan gene. These mice are viable and fertile and have no obvious deficits in reproduction and general performance. Brain anatomy, morphology, and ultrastructure are similar to those of wild-type mice. Perineuronal nets surrounding neurons appear largely normal. Mild deficits in synaptic plasticity may exist, as maintenance of late-phase hippocampal long term potentiation is reduced. These data indicate that neurocan has either a redundant or a more subtle function in the development of the brain. PMID- 11486036 TI - Regulation of transcription factor YY1 by acetylation and deacetylation. AB - YY1 is a sequence-specific DNA-binding transcription factor that has many important biological roles. It activates or represses many genes during cell growth and differentiation and is also required for the normal development of mammalian embryos. Previous studies have established that YY1 interacts with histone acetyltransferases p300 and CREB-binding protein (CBP) and histone deacetylase 1 (HDAC1), HDAC2, and HDAC3. Here, we present evidence that the activity of YY1 is regulated through acetylation by p300 and PCAF and through deacetylation by HDACs. YY1 was acetylated in two regions: both p300 and PCAF acetylated the central glycine-lysine-rich domain of residues 170 to 200, and PCAF also acetylated YY1 at the C-terminal DNA-binding zinc finger domain. Acetylation of the central region was required for the full transcriptional repressor activity of YY1 and targeted YY1 for active deacetylation by HDACs. However, the C-terminal region of YY1 could not be deacetylated. Rather, the acetylated C-terminal region interacted with HDACs, which resulted in stable HDAC activity associated with the YY1 protein. Finally, acetylation of the C-terminal zinc finger domain decreased the DNA-binding activity of YY1. Our findings suggest that in the natural context, YY1 activity is regulated through intricate mechanisms involving negative feedback loops, histone deacetylation, and recognition of the cognate DNA sequence affected by acetylation and deacetylation of the YY1 protein. PMID- 11486037 TI - Histone deacetylase 4 possesses intrinsic nuclear import and export signals. AB - Nucleocytoplasmic trafficking of histone deacetylase 4 (HDAC4) plays an important role in regulating its function, and binding of 14-3-3 proteins is necessary for its cytoplasmic retention. Here, we report the identification of nuclear import and export sequences of HDAC4. While its N-terminal 118 residues modulate the nuclear localization, residues 244 to 279 constitute an authentic, strong nuclear localization signal. Mutational analysis of this signal revealed that three arginine-lysine clusters are necessary for its nuclear import activity. As for nuclear export, leucine-rich sequences located in the middle part of HDAC4 do not function as nuclear export signals. By contrast, a hydrophobic motif (MXXLXVXV) located at the C-terminal end serves as a nuclear export signal that is necessary for cytoplasmic retention of HDAC4. This motif is required for CRM1-mediated nuclear export of HDAC4. Furthermore, binding of 14-3-3 proteins promotes cytoplasmic localization of HDAC4 by both inhibiting its nuclear import and stimulating its nuclear export. Unlike wild-type HDAC4, a point mutant with abrogated MEF2-binding ability remains cytoplasmic upon exogenous expression of MEF2C, supporting the notion that direct MEF2 binding targets HDAC4 to the nucleus. Therefore, HDAC4 possesses intrinsic nuclear import and export signals for its dynamic nucleocytoplasmic shuttling, and association with 14-3-3 and MEF2 proteins affects such shuttling and thus directs HDAC4 to the cytoplasm and the nucleus, respectively. PMID- 11486038 TI - Mre11 complex and DNA replication: linkage to E2F and sites of DNA synthesis. AB - We show that the Mre11 complex associates with E2F family members via the Nbs1 N terminus. This association and Nbs1 phosphorylation are correlated with S-phase checkpoint proficiency, whereas neither is sufficient individually for checkpoint activation. The Nbs1 E2F interaction occurred near the Epstein-Barr virus origin of replication as well as near a chromosomal replication origin in the c-myc promoter region and was restricted to S-phase cells. The Mre11 complex colocalized with PCNA at replication forks throughout S phase, both prior to and coincident with the appearance of nascent DNA. These data suggest that the Mre11 complex suppresses genomic instability through its influence on both the regulation and progression of DNA replication. PMID- 11486039 TI - Selective inactivation of p53 facilitates mouse epithelial tumor progression without chromosomal instability. AB - We examined the selective pressure for, and the impact of, p53 inactivation during epithelial tumor evolution in a transgenic brain tumor model. In TgT(121) mice, cell-specific inactivation of the pRb pathway in brain choroid plexus epithelium initiates tumorigenesis and induces p53-dependent apoptosis. We previously showed that p53 deficiency accelerates tumor growth due to diminished apoptosis. Here we show that in a p53(+/-) background, slow-growing dysplastic tissue undergoes clonal progression to solid angiogenic tumors in all animals. p53 is inactivated in all progressed tumors, with loss of the wild-type allele occurring in 90% of tumors. Moreover, similar progression occurs in 38% of TgT(121)p53(+/+) mice, also with loss of at least one p53 allele and inactivation of p53. Thus, the selective pressure for p53 inactivation, likely based on its apoptotic function, is high. Yet, in all cases, p53 inactivation correlates with progression beyond apoptosis reduction, from dysplasia to solid vascularized tumors. Hence, p53 suppresses tumor progression in this tissue by multiple mechanisms. Previous studies of fibroblasts and hematopoietic cells show that p53 deficiency can be associated with chromosomal instability, a mechanism that may drive tumor progression. To determine whether genomic gains or losses are present in tumors that progress in the absence of p53, we performed comparative genomic hybridization analysis. Surprisingly, the only detectable chromosomal imbalance was partial or complete loss of chromosome 11, which harbors the p53 gene and is thus the selected event. Flow cytometry confirmed that the majority of tumor cells were diploid. These studies indicate that loss of p53 function is frequent under natural selective pressures and furthermore that p53 loss can facilitate epithelial tumor progression by a mechanism in addition to apoptosis reduction and distinct from chromosomal instability. PMID- 11486040 TI - Impaired motor coordination in mice that lack punc. AB - The punc gene, encoding a member of the neural cell adhesion molecule family expressed in the developing central nervous system, limbs, and inner ear, was identified. To extend studies of the normal expression pattern of punc and to determine its function, a mouse strain bearing a lacZ/neo insertion in a 5' coding exon was created. The complex pattern of punc expression in embryos from embryonic day 9.5 (E9.5) to E11.5 was mimicked accurately by beta-galactosidase (beta-Gal) activity. As development proceeded, the distribution of beta-Gal activity was increasingly restricted, finally becoming confined to the brain and inner ear by E15.5. In the adult, beta-Gal activity was detected in several regions of the inner ear and brain and was particularly strong in the cerebellar Bergmann glia. Genetic analysis of this null allele demonstrated that punc is not required for normal embryogenesis. Interestingly, comparisons of beta-Gal activity and punc transcripts in heterozygous and homozygous mutant individuals demonstrated that punc is negatively autoregulated in some tissues. Adult punc deficient mice were overtly normal and had normal hearing. Compared with control littermates, however, homozygous mutants had significantly reduced retention times on the Rotarod, suggesting a role for Bergmann glia-expressed Punc in the cerebellar control of motor coordination. PMID- 11486041 TI - Intact lysosome transport and phagosome function despite kinectin deficiency. AB - The mechanism of cargo coupling to kinesin motor proteins is a fundamental issue in organelle transport along microtubules. Kinectin has been postulated to function as a membrane anchor protein that attaches various organelles to the prototype motor protein kinesin. To verify the biological relevance of kinectin in vivo, the murine kinectin gene was disrupted by homologous recombination. Unexpectedly, kinectin-deficient mice were viable and fertile, and no gross abnormalities were observed up to 1 year of age. The assembly of the endoplasmic reticulum was essentially unaffected in kinectin-deficient cells. Mitochondria appeared to be correctly distributed throughout the cytoplasm along the microtubules. Furthermore, the stationary distribution and the bidirectional movement of lysosomes did not depend on kinectin. Kinectin-deficient phagocytes internalized and cleared bacteria, indicating that phagosome trafficking and maturation are functional without kinectin. Thus, these data unequivocally indicate that kinectin is not essential for trafficking of lysosomes, phagosomes, and mitochondria in vivo. PMID- 11486042 TI - Partners of Rpb8p, a small subunit shared by yeast RNA polymerases I, II and III. AB - Rpb8p, a subunit common to the three yeast RNA polymerases, is conserved among eukaryotes and absent from noneukaryotes. Defective mutants were found at an invariant GGLLM motif and at two other highly conserved amino acids. With one exception, they are clustered on the Rpb8p structure. They all impair a two hybrid interaction with a fragment conserved in the largest subunits of RNA polymerases I (Rpa190p), II (Rpb1p), and III (Rpc160p). This fragment corresponds to the pore 1 module of the RNA polymerase II crystal structure and bears a highly conserved motif (P.I.KP.LW.GKQ) facing the GGLLM motif of Rpb8p. An RNA polymerase I mutant (rpa190-G728D) at the invariant glycyl of P.I.KP.LW.GKQ provokes a temperature-sensitive defect. Increasing the gene dosage of another common subunit, Rpb6p, suppresses this phenotype. It also suppresses a conditional growth defect observed when replacing Rpb8p by its human counterpart. Hence, Rpb6p and Rpb8p functionally interact in vivo. These two subunits are spatially separated by the pore 1 module and may also be possibly connected by the disorganized N half of Rpb6p, not included in the present structure data. Human Rpb6p is phosphorylated at its N-terminal Ser2, but an alanyl replacement at this position still complements an rpb6-Delta null allele. A two-hybrid interaction also occurs between Rpb8p and the product of orphan gene YGR089w. A ygr089-Delta null mutant has no detectable growth defect but aggravates the conditional growth defect of rpb8 mutants, suggesting that the interaction with Rpb8p may be physiologically relevant. PMID- 11486043 TI - Spermatogenesis and the regulation of Ca(2+)-calmodulin-dependent protein kinase IV localization are not dependent on calspermin. AB - Calspermin and Ca(2+)-calmodulin-dependent protein kinase IV (CaMKIV) are two proteins encoded by the Camk4 gene. CaMKIV is found in multiple tissues, including brain, thymus, and testis, while calspermin is restricted to the testis. In the mouse testis, both proteins are expressed within elongating spermatids. We have recently shown that deletion of CaMKIV has no effect on calspermin expression but does impair spermiogenesis by disrupting the exchange of sperm basic nuclear proteins. The function of calspermin within the testis is unclear, although it has been speculated to play a role in binding and sequestering calmodulin during the development of the germ cell. To investigate the contribution of calspermin to spermatogenesis, we have used Cre/lox technology to specifically delete calspermin, while leaving kinase expression intact. We unexpectedly found that calspermin is not required for male fertility. We further demonstrate that CaMKIV expression and localization are unaffected by the absence of calspermin and that calspermin does not colocalize to the nuclear matrix with CaMKIV. PMID- 11486044 TI - HERP, a new primary target of Notch regulated by ligand binding. AB - Notch signaling dictates cell fate and critically influences cell proliferation, differentiation, and apoptosis in metazoans. Ligand binding initiates the signal through regulated intramembrane proteolysis of a transmembrane Notch receptor which releases the signal-transducing Notch intracellular domain (NICD). The HES/E(spl) gene family is a primary target of Notch and thus far the only known Notch effector. A newly isolated HERP family, a HES-related basic helix-loop helix protein family, has been proposed as a potential target of Notch, based on its induction following NICD overexpression. However, NICD is physiologically maintained at an extremely low level that typically escapes detection, and therefore, nonregulated overexpression of NICD-as in transient transfection-has the potential of generating cellular responses of little physiological relevance. Indeed, a constitutively active NICD indiscriminately up-regulates expression of both HERP1 and HERP2 mRNAs. However, physiological Notch stimulation through ligand binding results in the selective induction of HERP2 but not HERP1 mRNA and causes only marginal up-regulation of HES1 mRNA. Importantly, HERP2 is an immediate target gene of Notch signaling since HERP2 mRNA expression is induced even in the absence of de novo protein synthesis. HERP2 mRNA induction is accompanied by specific expression of HERP2 protein in the nucleus. Furthermore, using RBP-Jk-deficient cells, we show that an RBP-Jk protein, a transcription factor that directly activates HES/E(spl) transcription, also is essential for HERP2 mRNA expression and that expression of exogenous RBP-Jk is sufficient to rescue HERP2 mRNA expression. These data establish that HERP2 is a novel primary target gene of Notch that, together with HES, may effect diverse biological activities of Notch. PMID- 11486046 TI - Oversight of genetic testing: an update. PMID- 11486045 TI - HERP, a novel heterodimer partner of HES/E(spl) in Notch signaling. AB - HERP1 and -2 are members of a new basic helix-loop-helix (bHLH) protein family closely related to HES/E(spl), the only previously known Notch effector. Like that of HES, HERP mRNA expression is directly up-regulated by Notch ligand binding without de novo protein synthesis. HES and HERP are individually expressed in certain cells, but they are also coexpressed within single cells after Notch stimulation. Here, we show that HERP has intrinsic transcriptional repression activity. Transcriptional repression by HES/E(spl) entails the recruitment of the corepressor TLE/Groucho via a conserved WRPW motif, whereas unexpectedly the corresponding-but modified-tetrapeptide motif in HERP confers marginal repression. Rather, HERP uses its bHLH domain to recruit the mSin3 complex containing histone deacetylase HDAC1 and an additional corepressor, N CoR, to mediate repression. HES and HERP homodimers bind similar DNA sequences, but with distinct sequence preferences, and they repress transcription from specific DNA binding sites. Importantly, HES and HERP associate with each other in solution and form a stable HES-HERP heterodimer upon DNA binding. HES-HERP heterodimers have both a greater DNA binding activity and a stronger repression activity than do the respective homodimers. Thus, Notch signaling relies on cooperation between HES and HERP, two transcriptional repressors with distinctive repression mechanisms which, either as homo- or as heterodimers, regulate target gene expression. PMID- 11486047 TI - Expression profiling of human tumors: the end of surgical pathology? PMID- 11486048 TI - Human leukocyte antigen gene polymorphism and the histocompatibility laboratory. AB - The human leukocyte antigens (HLA) encoded by genes within the major histocompatibility complex display an impressive degree of polymorphism. This variability is apparently maintained in human populations through the need to successfully display a wide range of processed foreign peptides to the T cell antigen receptor. The large number of alleles at the Class I and Class II loci pose a significant problem for molecular diagnosis. Knowledge of allele groups and specific alleles present in individuals has important implications in organ and stem cell transplantation and in disease association studies. Histocompatibility laboratories have transformed themselves during the past decade as they have adapted the techniques of molecular diagnostics to the challenge of identifying HLA alleles. PMID- 11486049 TI - Laser-assisted microdissection of membrane-mounted paraffin sections for polymerase chain reaction analysis: identification of cell populations using immunohistochemistry and in situ hybridization. AB - Laser microbeam microdissection (LMM) is an increasingly important method for obtaining pure cell samples for genetic and proteomic analysis. Immunohistochemistry (IHC) and in situ hybridization (ISH) are useful techniques for targeting specific cell populations for microdissection but are difficult to apply with the tissue support membranes often used during LMM. Using detection of cytokeratins and Epstein-Barr virus gene products in head and neck carcinoma as a model, we describe optimized protocols for membrane and section preparation and for low temperature antigen retrieval that allow IHC and ISH to be used reliably on membrane mounted paraffin tissue sections. Visualization of cellular targets was markedly improved by staining and this could be further improved using a variety of optical media before microdissection. Tissue fragments thus stained were suitable for subsequent polymerase chain reaction analysis of extracted DNA using standard techniques. These IHC and ISH procedures are generally applicable and will be useful for detecting a wide range of antigens and nucleic acids in paraffin sections in conjunction with LMM. PMID- 11486051 TI - t(15;17) Reverse transcription-polymerase chain reaction with alternative splicing. PMID- 11486050 TI - Simultaneous quantification of human glandular kallikrein 2 and prostate-specific antigen mRNAs in peripheral blood from prostate cancer patients. AB - We present a multiplexed and internally calibrated quantitative reverse transcription-PCR (QRT-PCR) assay to detect human glandular kallikrein 2 (hK2) and prostate-specific antigen (PSA) transcripts in blood samples from healthy subjects and prostate cancer (PC) patients. The assay detected 50 copies of hK2 and PSA mRNA, and 1 PSA- and 10 hK2-expressing LNCaP cells in the presence of 2.5 x 10(6) PSA- and hK2-negative cells. In PC patients, 20 of 25 and 19 of 25 gave detectable PSA and hK2 mRNAs, respectively. Number of hK2 mRNA copies was significantly higher than that of PSA mRNA copies in patients with biochemically progressive (P = 0.02) PC, and with locally advanced and metastasized (P = 0.004) PC. Patients with rapidly progressive and hormone refractory PC gave detectable hK2 mRNA only in 2 of 8 and PSA mRNA in 3 of 8 patients. Neither PSA nor hK2 mRNAs were detected in 16 healthy subjects. PSA and hK2 discriminated PC patients with biochemically progressive and advanced disease from the controls and from the aggressive distant metastatic disease. The assay provides a reliable quantification of the number of hK2 and PSA mRNA copies, allows to discriminate PC cases from healthy subjects, and offers a tool for further studies on molecular staging of PC. PMID- 11486052 TI - Highlights of the 10th Annual Beaumont Symposium on Molecular Pathology: "DNA Technology in the Clinical Laboratory". March 8-10, 2001. PMID- 11486053 TI - A role for the RNase III enzyme DCR-1 in RNA interference and germ line development in Caenorhabditis elegans. AB - An early event in RNA interference (RNAi) is the cleavage of the initiating double-stranded RNA (dsRNA) to short pieces, 21 to 23 nucleotides in length. Here we describe a null mutation in dicer-1 (dcr-1), a gene proposed to encode the enzyme that generates these short RNAs. We find that dcr-1(-/-) animals have defects in RNAi under some, but not all, conditions. Mutant animals have germ line defects that lead to sterility, suggesting that cleavage of dsRNA to short pieces is a requisite event in normal development. PMID- 11486054 TI - Patterns of gene expression during Drosophila mesoderm development. AB - The transcription factor Twist initiates Drosophila mesoderm development, resulting in the formation of heart, somatic muscle, and other cell types. Using a Drosophila embryo sorter, we isolated enough homozygous twist mutant embryos to perform DNA microarray experiments. Transcription profiles of twist loss-of function embryos, embryos with ubiquitous twist expression, and wild-type embryos were compared at different developmental stages. The results implicate hundreds of genes, many with vertebrate homologs, in stage-specific processes in mesoderm development. One such gene, gleeful, related to the vertebrate Gli genes, is essential for somatic muscle development and sufficient to cause neural cells to express a muscle marker. PMID- 11486056 TI - Memory consolidation and NMDA receptors: discrepancy between genetic and pharmacological approaches. PMID- 11486055 TI - Skinny hedgehog, an acyltransferase required for palmitoylation and activity of the hedgehog signal. AB - One of the most dominant influences in the patterning of multicellular embryos is exerted by the Hedgehog (Hh) family of secreted signaling proteins. Here, we identify a segment polarity gene in Drosophila melanogaster, skinny hedgehog (ski), and show that its product is required in Hh-expressing cells for production of appropriate signaling activity in embryos and in the imaginal precursors of adult tissues. The ski gene encodes an apparent acyltransferase, and we provide genetic and biochemical evidence that Hh proteins from ski mutant cells retain carboxyl-terminal cholesterol modification but lack amino-terminal palmitate modification. Our results suggest that ski encodes an enzyme that acts within the secretory pathway to catalyze amino-terminal palmitoylation of Hh, and further demonstrate that this lipid modification is required for the embryonic and larval patterning activities of the Hh signal. PMID- 11486057 TI - Good news, bad news. PMID- 11486058 TI - Volcanology. Etna eruption puts volcano monitoring to the test. PMID- 11486059 TI - Stem cells. Japan readies rules that allow research. PMID- 11486060 TI - Earth sciences. Satellite shutdown stirs controversy. PMID- 11486061 TI - Heavy-ion physics. Berkeley crew unbags element 118. PMID- 11486062 TI - Entomology. First light on genetic roots of Bt resistance. PMID- 11486063 TI - Paleontology. Dinosaur nostrils get a hole new look. PMID- 11486064 TI - Genetic engineering. Imperial College fined over hybrid virus risk. PMID- 11486065 TI - National security. MIT military critic rejects secrecy claims. PMID- 11486066 TI - Nanocomputing. Assembling nanocircuits from the bottom up. PMID- 11486067 TI - Nanocomputing. Yet another role for DNA? PMID- 11486068 TI - Nanocomputing. Optical lithography goes to extremes--and beyond. PMID- 11486069 TI - Nanocomputing. World's smallest transistor. PMID- 11486070 TI - Nanocomputing. The end--not here yet, but coming soon. PMID- 11486071 TI - International Congress of Developmental Biology. Developmental progress fills the air in Kyoto. PMID- 11486073 TI - Mathematics. Randomly distributed slices of pi. PMID- 11486072 TI - Peter Brewer. Fathoming the chemistry of the deep blue sea. PMID- 11486074 TI - Essays on science and society. Defining disease in the genomics era. PMID- 11486076 TI - Geophysics. Deep diamond mysteries. PMID- 11486075 TI - Development. Dicing up RNAs. PMID- 11486077 TI - Astronomy. The complex story of H2. PMID- 11486078 TI - Biomedicine. Reconstructing myotonic dystrophy. PMID- 11486079 TI - Protein synthesis. The perks of balancing glucose. AB - What do the regulation of translation initiation and glucose metabolism have to do with each other? Quite a lot, it seems, according to Sonenberg and Newgard in their Perspective. They discuss new findings that identify the kinase responsible for inactivating eIF2--a factor that is required for translation initiation (and hence protein synthesis)--when the endoplasmic reticulum is under stress. Loss of this kinase results in destruction of insulin-producing b cells in the pancreas and dysregulation of glucose homeostasis. PMID- 11486080 TI - Earth history. The rise of atmospheric oxygen. PMID- 11486082 TI - Biogenic methane, hydrogen escape, and the irreversible oxidation of early Earth. AB - The low O2 content of the Archean atmosphere implies that methane should have been present at levels approximately 10(2) to 10(3) parts per million volume (ppmv) (compared with 1.7 ppmv today) given a plausible biogenic source. CH4 is favored as the greenhouse gas that countered the lower luminosity of the early Sun. But abundant CH4 implies that hydrogen escapes to space (upward arrow space) orders of magnitude faster than today. Such reductant loss oxidizes the Earth. Photosynthesis splits water into O2 and H, and methanogenesis transfers the H into CH4. Hydrogen escape after CH4 photolysis, therefore, causes a net gain of oxygen [CO2 + 2H2O --> CH4 + 2O2 --> CO2 + O2 + 4H(upward arrow space)]. Expected irreversible oxidation (approximately 10(12) to 10(13) moles oxygen per year) may help explain how Earth's surface environment became irreversibly oxidized. PMID- 11486081 TI - Induction of apoptosis by a secreted lipocalin that is transcriptionally regulated by IL-3 deprivation. AB - Many hematopoietic cells undergo apoptosis when deprived of specific cytokines, and this process requires de novo RNA/protein synthesis. Using DNA microarrays to analyze interleukin-3 (IL-3)-dependent murine FL5.12 pro-B cells, we found that the gene undergoing maximal transcriptional induction after cytokine withdrawal is 24p3, which encodes a secreted lipocalin. Conditioned medium from IL-3 deprived FL5.12 cells contained 24p3 and induced apoptosis in naive FL5.12 cells even when IL-3 was present. 24p3 also induced apoptosis in a wide variety of leukocytes but not other cell types. Apoptotic sensitivity correlated with the presence of a putative 24p3 cell surface receptor. We conclude that IL-3 deprivation activates 24p3 transcription, leading to synthesis and secretion of 24p3, which induces apoptosis through an autocrine pathway. PMID- 11486083 TI - Josephson junction arrays with Bose-Einstein condensates. AB - We report on the direct observation of an oscillating atomic current in a one dimensional array of Josephson junctions realized with an atomic Bose-Einstein condensate. The array is created by a laser standing wave, with the condensates trapped in the valleys of the periodic potential and weakly coupled by the interwell barriers. The coherence of multiple tunneling between adjacent wells is continuously probed by atomic interference. The square of the small-amplitude oscillation frequency is proportional to the microscopic tunneling rate of each condensate through the barriers and provides a direct measurement of the Josephson critical current as a function of the intermediate barrier heights. Our superfluid array may allow investigation of phenomena so far inaccessible to superconducting Josephson junctions and lays a bridge between the condensate dynamics and the physics of discrete nonlinear media. PMID- 11486084 TI - Soluble and colloidal iron in the oligotrophic North Atlantic and North Pacific. AB - In the oligotrophic North Atlantic and North Pacific, ultrafiltration studies show that concentrations of soluble iron and soluble iron-binding organic ligands are much lower than previously presumed "dissolved" concentrations, which were operationally defined as that passing through a 0.4-micrometer pore filter. Our studies indicate that substantial portions of the previously presumed "dissolved" iron (and probably also iron-binding ligands) are present in colloidal size range. The soluble iron and iron-binding organic ligands are depleted at the surface and enriched at depth, similar to distributions of major nutrients. By contrast, colloidal iron shows a maximum at the surface and a minimum in the upper nutricline. Our results suggest that "dissolved" iron may be less bioavailable to phytoplankton than previously thought and that iron removal through colloid aggregation and settling should be considered in models of the oceanic iron cycle. PMID- 11486085 TI - Nostril position in dinosaurs and other vertebrates and its significance for nasal function. AB - Many dinosaurs have enormous and complicated bony nasal apertures. Functional interpretation requires knowledge of the location of the external opening in the skin. Traditionally, the fleshy nostril of dinosaurs has been placed in the back of the bony opening, but studies of extant dinosaur relatives suggest that it is located far forward. Narial blood supply and cavernous tissue corroborate the rostral position in dinosaurs. A rostral nostril was, and remains, a virtually invariant rule of construction among Amniota, which has consequences for (i) nasal airstreaming, and hence various physiological parameters, and (ii) the collection of behaviorally relevant circumoral odorants. PMID- 11486086 TI - Identification of a gene associated with Bt resistance in Heliothis virescens. AB - Transgenic crops producing insecticidal toxins from Bacillus thuringiensis (Bt) are widely used for pest control. Bt-resistant insect strains have been studied, but the molecular basis of resistance has remained elusive. Here, we show that disruption of a cadherin-superfamily gene by retrotransposon-mediated insertion was linked to high levels of resistance to the Bt toxin Cry1Ac in the cotton pest Heliothis virescens. Monitoring the early phases of Bt resistance evolution in the field has been viewed as crucial but extremely difficult, especially when resistance is recessive. Our findings enable efficient DNA-based screening for resistant heterozygotes by directly detecting the recessive allele. PMID- 11486087 TI - Bt toxin resistance from loss of a putative carbohydrate-modifying enzyme. AB - The development of resistance is the main threat to the long-term use of toxins from Bacillus thuringiensis (Bt) in transgenic plants. Here we report the cloning of a Bt toxin resistance gene, Caenorhabditis elegans bre-5, which encodes a putative beta-1,3-galactosyltransferase. Lack of bre-5 in the intestine led to resistance to the Bt toxin Cry5B. Wild-type but not bre-5 mutant animals were found to uptake toxin into their gut cells, consistent with bre-5 mutants lacking toxin-binding sites on their apical gut. bre-5 mutants displayed resistance to Cry14A, a Bt toxin lethal to both nematodes and insects; this indicates that resistance by loss of carbohydrate modification is relevant to multiple Bt toxins. PMID- 11486088 TI - Myotonic dystrophy type 2 caused by a CCTG expansion in intron 1 of ZNF9. AB - Myotonic dystrophy (DM), the most common form of muscular dystrophy in adults, can be caused by a mutation on either chromosome 19q13 (DM1) or 3q21 (DM2/PROMM). DM1 is caused by a CTG expansion in the 3' untranslated region of the dystrophia myotonica-protein kinase gene (DMPK). Several mechanisms have been invoked to explain how this mutation, which does not alter the protein-coding portion of a gene, causes the specific constellation of clinical features characteristic of DM. We now report that DM2 is caused by a CCTG expansion (mean approximately 5000 repeats) located in intron 1 of the zinc finger protein 9 (ZNF9) gene. Parallels between these mutations indicate that microsatellite expansions in RNA can be pathogenic and cause the multisystemic features of DM1 and DM2. PMID- 11486089 TI - Stereotyped position of local synaptic targets in neocortex. AB - The microcircuitry of the mammalian neocortex remains largely unknown. Although the neocortex could be composed of scores of precise circuits, an alternative possibility is that local connectivity is probabilistic or even random. To examine the precision and degree of determinism in the neocortical microcircuitry, we used optical probing to reconstruct microcircuits in layer 5 from mouse primary visual cortex. We stimulated "trigger" cells, isolated from a homogenous population of corticotectal pyramidal neurons, while optically detecting "follower" neurons directly driven by the triggers. Followers belonged to a few selective anatomical classes with stereotyped physiological and synaptic responses. Moreover, even the position of the followers appeared determined across animals. Our data reveal precisely organized cortical microcircuits. PMID- 11486090 TI - Sorting of striatal and cortical interneurons regulated by semaphorin-neuropilin interactions. AB - Most striatal and cortical interneurons arise from the basal telencephalon, later segregating to their respective targets. Here, we show that migrating cortical interneurons avoid entering the striatum because of a chemorepulsive signal composed at least in part of semaphorin 3A and semaphorin 3F. Migrating interneurons expressing neuropilins, receptors for semaphorins, are directed to the cortex; those lacking them go to the striatum. Loss of neuropilin function increases the number of interneurons that migrate into the striatum. These observations reveal a mechanism by which neuropilins mediate sorting of distinct neuronal populations into different brain structures, and provide evidence that, in addition to guiding axons, these receptors also control neuronal migration in the central nervous system. PMID- 11486091 TI - Reciprocal regulation between TOC1 and LHY/CCA1 within the Arabidopsis circadian clock. AB - The interactive regulation between clock genes is central for oscillator function. Here, we show interactions between the Arabidopsis clock genes LATE ELONGATED HYPOCOTYL (LHY), CIRCADIAN CLOCK ASSOCIATED 1 (CCA1), and TIMING OF CAB EXPRESSION 1 (TOC1). The MYB transcription factors LHY and CCA1 negatively regulate TOC1 expression. We show that both proteins bind to a region in the TOC1 promoter that is critical for its clock regulation. Conversely, TOC1 appears to participate in the positive regulation of LHY and CCA1 expression. Our results indicate that these interactions form a loop critical for clock function in Arabidopsis. PMID- 11486092 TI - 2001: A prospective, seasonal odyssey into antiphospholipid protein antibodies. PMID- 11486093 TI - beta(2)-Glycoprotein 1-dependent anticardiolipin antibodies and risk of ischemic stroke and myocardial infarction: the honolulu heart program. AB - BACKGROUND: It has been hypothesized that immunoreactivity to beta(2) glycoprotein 1 (beta2GP1)-dependent anticardiolipin antibody (aCL), but not beta2GP1-independent aCL, is associated with increased risk of ischemic stroke and myocardial infarction (MI). METHODS: We performed a nested case-control study examining aCL as a risk factor for ischemic stroke and MI by using stored frozen sera obtained from subjects enrolled in the Honolulu Heart Program and followed for up for 20 years. We measured beta2GP1-dependent and beta2GP1-independent aCL and anti-beta2GP1 immunoreactivity in 259 men who developed an ischemic stroke, in 374 men who developed an MI, and in a control group of 1360 men who remained free of both conditions. RESULTS: Only beta2GP1-dependent aCL of the IgG class was significantly associated with both incident ischemic stroke and MI. This association was attenuated in the last 5 years of the 20-year follow-up. For stroke, the risk factor-adjusted relative odds for men with a positive versus a negative beta2GP1-dependent aCL of the IgG class were 2.2 (95% CI 1.5 to 3.4) at 15 years and 1.5 (95% CI 1.0 to 2.3) at 20 years. For MI, the adjusted relative odds were 1.8 (95% CI 1.2 to 2.6) at 15 years and 1.5 (95% CI 1.1 to 2.1) at 20 years. CONCLUSIONS: These data suggest that aCL IgG, particularly the beta2GP1 dependent variety, is an important predictor of future stroke and MI in men. PMID- 11486094 TI - Seasonal distribution of antiphospholipid antibodies. AB - BACKGROUND AND PURPOSE: The purpose of this study was to determine if there was a seasonal variation in antiphospholipid antibody (aPL) titers and whether this variation differed between stroke cases and control subjects. METHODS: IgG and IgM anticardiolipin and antiphosphatidyl serine antibody titers were obtained on serum samples from 884 stroke patients and 1024 control subjects over a 7-year period. Temporal distributions by month of blood draw were evaluated. RESULTS: Marked seasonal differences in the proportion of positive titers were found for control subjects, but no seasonal variability among patients was noted. In control subjects, positive titers occurred less frequently in the summer months, mirroring the seasonal trends seen in respiratory track infections and rheumatic fever. CONCLUSIONS: Our data suggest some aPL antibodies arise from different origins in patients and control subjects. The seasonality observed in the apparently normal population may be related to antibodies of infectious origin and is consistent with the reported lack of association with thrombosis of infection-related antibodies. PMID- 11486095 TI - Adaptive behavior in stressful situations and stroke incidence in hypertensive men: results from prospective cohort study "men born in 1914" in Malmo, Sweden. AB - BACKGROUND AND PURPOSE: Although hypertension is a major risk factor for stroke, many hypertensive persons remain healthy. The aim of the present study was to analyze whether adaptation in a stressful situation was associated with the incidence of stroke in hypertensive men. METHODS: Two hundred thirty-eight hypertensive men were followed from baseline in 1982/1983 until first stroke, death, or December 31, 1996. Adaptation to stress was studied with the serial Color-Word Test. In the Regression dimension, 4 patterns of adaptation could be distinguished according to mastering of the test. Successful mastering of the test was shown in stabilized patterns, increasing difficulty in cumulative patterns, fluctuating difficulty in dissociative patterns, and fluctuating difficulty that increased during testing in cumulative-dissociative patterns. The patterns were compared regarding stroke incidence. RESULTS: Forty-three men experienced a stroke during follow-up. Stroke rates per 1000 person-years were 12.6 for men with stabilized patterns, 14.3 for men with cumulative patterns, 16.2 for men with dissociative patterns, and 31.2 for men with cumulative dissociative patterns. Multivariate analysis, adjusted for relevant cerebrovascular risk factors, showed that the cumulative-dissociative pattern of the Regression dimension was associated with an increased risk of stroke during follow-up (relative risk 3.00, 95% CI 1.32 to 6.81). CONCLUSIONS: The specific behavior pattern, characterized by the greatest difficulties in managing the test, was associated with incidence of stroke in hypertensive men. One interpretation is that hypertensive men who chronically fail to find successful strategies in stressful situations are vulnerable to the damaging effects of stress and thereby at an increased risk of a future stroke. PMID- 11486096 TI - Ankle-brachial index and 7-year ischemic stroke incidence: the ARIC study. AB - BACKGROUND AND PURPOSE: Low ankle-brachial index (ABI), which is the ratio of tibial artery systolic blood pressure to brachial systolic artery pressure, is known to be a measure of lower limb peripheral artery disease as well as a marker for other cardiovascular disease events. The ability of ABI to predict incident ischemic stroke, however, is not established in population-based studies. METHODS: ABI was measured in a cohort of 14 839 black and white men and women aged 45 to 64 years. Stroke incidence was calculated during approximately 7 years of follow-up. RESULTS: A total of 206 incident strokes occurred. Adjusted stroke incidence rates were markedly higher for those in the lowest versus the highest categories of ABI for men, women, blacks, and whites. The proportional hazards regression model, adjusted for age, race, gender, and field center, showed an inverse linear trend between ABI and ischemic stroke incidence (P<0.0001). The lowest group (ABI <0.80) had a hazard ratio of 5.68 (95% CI 2.77 to 11.66). After adjustment for major risk factors in a multivariate model, the hazard ratio in the lowest group was elevated (1.93) but no longer statistically significant (95% CI 0.78 to 4.78). There was, however, still an indication of an overall inverse linear trend between ABI and incident stroke (P=0.03). CONCLUSIONS: Low ABI was strongly associated with increased incidence of ischemic stroke, but the relationship was substantially reduced after adjustment for major cardiovascular risk factors. PMID- 11486097 TI - Race-ethnic disparities in the impact of stroke risk factors: the northern Manhattan stroke study. AB - BACKGROUND AND PURPOSE: Stroke risk factors have been determined in large part through epidemiological studies in white cohorts; as a result, race-ethnic disparities in stroke incidence and mortality rates remained unexplained. The aim in the present study was to compare the prevalence, OR, and etiological fraction (EF) of stroke risk factors among white, blacks, and Caribbean Hispanics living in the same urban community of northern Manhattan. METHODS: In this population based incident case-control study, cases (n=688) of first ischemic stroke were prospectively matched 1:2 by age, sex, and race-ethnicity with community controls (n=1156). Risk factors were determined through in-person assessment. Conditional logistic regression was used to calculate adjusted ORs in each race-ethnic group. Prevalence and multivariate EFs were determined in each race-ethnic group. RESULTS: Hypertension was an independent risk factor for whites (OR 1.8, EF 25%), blacks (OR 2.0, EF 37%), and Caribbean Hispanics (OR 2.1, EF 32%), but greater prevalence led to elevated EFs among blacks and Caribbean Hispanics. Greater prevalence rates of diabetes increased stroke risk in blacks (OR 1.8, EF 14%) and Caribbean Hispanics (OR 2.1 P<0.05, EF 10%) compared with whites (OR 1.0, EF 0%), whereas atrial fibrillation had a greater prevalence and EF for whites (OR 4.4, EF 20%) compared with blacks (OR 1.7, EF 3%) and Caribbean Hispanics (OR 3.0, EF 2%). Coronary artery disease was most important for whites (OR 1.3, EF 16%), followed by Caribbean Hispanics (OR 1.5, EF 6%) and then blacks (OR 1.1, EF 2%). Prevalence of physical inactivity was greater in Caribbean Hispanics, but an elevated EF was found in all groups. CONCLUSIONS: The prevalence, OR, and EF for stroke risk factors vary by race-ethnicity. These differences are crucial to the etiology of stroke, as well as to the design and implementation of stroke prevention programs. PMID- 11486098 TI - Incidence of the major stroke subtypes: initial findings from the North East Melbourne stroke incidence study (NEMESIS). AB - BACKGROUND AND PURPOSE: Population-based stroke incidence studies are the only accurate way to determine the number of strokes that occur in a given society. Because the major stroke subtypes have different patterns of incidence and outcome, information on the natural history of stroke subtypes is essential. The purpose of the present study was to determine the incidence and case-fatality rate of the major stroke subtypes in a geographically defined region of Melbourne, Australia. METHODS: All suspected strokes that occurred among 133 816 residents of suburbs north and east of Melbourne, Australia, during a 12-month period of 1996 and 1997 were identified and assessed. Multiple overlapping sources were used to ascertain cases, and standard criteria for stroke and case fatality were used. Stroke subtypes were defined by CT, MRI, and autopsy. RESULTS: Three hundred eighty-one strokes occurred among 353 persons during the study period, with 276 (72%) being first-ever-in-a-lifetime strokes. Of these, 72.5% (95% CI 67.2% to 77.7%) were cerebral infarction, 14.5% (95% CI 10.3% to 18.6%) were intracerebral hemorrhage, 4.3% (95% CI 1.9% to 6.8%) were subarachnoid hemorrhage, and 8.7% (95% CI 5.4% to 12.0%) were stroke of undetermined type. The 28-day case-fatality rate was 12% (95% CI 7% to 16%) for cerebral infarction, 45% (95% CI 30% to 60%) for intracerebral hemorrhage, 50% (95% CI 22% to 78%) for subarachnoid hemorrhage, and 38% (95% CI 18% to 57%) for stroke of undetermined type. CONCLUSIONS: The overall distribution of stroke subtypes and 28-day case-fatality rates are not significantly different from those of most European countries or the United States. There may, however, be some differences in the incidence of subtypes within Australia. PMID- 11486099 TI - Life expectancies among survivors of acute cerebrovascular disease. AB - BACKGROUND AND PURPOSE: Stroke survivors represent a large group of persons for whom age-differentiated life expectancy tables do not exist. Such tables are vital for many purposes. The aim of the present study was to estimate age- and sex-specific life expectancies among individuals who have survived the acute phase (1 month) of a cerebrovascular disease (CVD). METHODS: All patients who were registered with the Swedish National Hospital Discharge Registry with an admission for CVD (ICD codes 430 to 438) between January 1, 1989, and November 30, 1993, and were alive at the end of 1993 (N=103 591) were followed for mortality rates in 1994. The same was done for 1983. Actuarial analyses were used to convert death rates into life expectancies. RESULTS: Life expectancy among CVD survivors increased with time (1983 versus 1994): 22.9% for men (95% CI 18.3% to 27.6%) and 12.9% for women (95% CI 9.1% to 16.6%). The life expectancy ratio in 1983 between CVD survivors and the general population was 0.571 (95% CI 0.533 to 0.590) for men and 0.578 (95% CI 0.562 to 0.592) for women. In 1994, the corresponding ratios were 0.641 (95% CI 0.629 to 0.654) and 0.611 (95% CI 0.601 to 0.622). The life expectancy ratios between female and male survivors were 1.28 (95% CI 1.23 to 1.34) in 1983 and 1.18 (95% CI 1.15 to 1.21) in 1994. The prognosis for survivors who experienced occlusion and stenosis of the precerebral arteries was better than that for survivors of an intracerebral hemorrhage (P=4.4E-4) or occlusion of cerebral arteries (P=3.8E-8). CONCLUSIONS: Although the prognosis has improved for all ages, stroke survivors still constitute a large group of persons with a low life expectancy compared with the general population. PMID- 11486100 TI - Changes in stroke mortality rates for 1950 to 1997: a great slowdown of decline trend in Japan. AB - BACKGROUND AND PURPOSE: Stroke is the second leading cause of death in Japan. We sought to examine changes in stroke mortality rates by age, sex, and birth cohorts from 1950 to 1997. METHODS: The numbers of deaths from stroke and the population by age and sex in subjects aged 55 to 79 years between 1950 and 1997 were obtained from national vital statistics. Poisson regression analyses for annual percentage changes were used to explore these trends. RESULTS: Stroke mortality rates decreased since the mid-1960s for both men and women. Four periods of the decline are identified. In the first period (1950 to 1964), the age-adjusted annual percent changes averaged 0.9% for men (P<0.05) and -0.6% for women (P>0.05). In the second period (1965 to 1974), the annual change averaged 5.0% for men and -5.2% for women (both P<0.01). In the third period (1975 to 1989), the annual change averaged -8.6% for men and -8.7% for women (both P<0.01). In the fourth period (1990 to 1997), the annual change averaged -1.2% for men (P>0.05) and -3.0% for women (P<0.01). In the fourth period, the slowdown of the decline in stroke mortality was most evident in the older age groups of men. CONCLUSIONS: The findings suggested that along with an increasing aging population, the slowdown in the decline of stroke mortality rates, especially for men, is of considerable concern. Efforts to control stroke should be vigorously continued in the 2000s. PMID- 11486101 TI - Evolution of cerebral tumor necrosis factor-alpha production during human ischemic stroke. AB - BACKGROUND AND PURPOSE: Tumor necrosis factor-alpha (TNF-alpha) is detected in ischemic brain cells in experimental animal models and is believed to play an important role in apoptosis. However, the natural expression of TNF-alpha during human stroke is not known. METHODS: We examined TNF-alpha immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) in brain samples of stroke victims (n=16) after variable survival (15 hours to 18 days). Systemic TNF-alpha content from a separate cohort including severe or lethal stroke cases (n=26) was also assayed. RESULTS: Neuronal TNF-alpha was demonstrated from 0.6 to 5.4 days after the onset of stroke symptoms, peaking bilaterally during days 2 and 3. Bilateral glial TNF-alpha immunoreactivity was detected during the acute phase, with the astrocytic TNF-alpha expression dominating in later phases and persisting contralaterally to the infarct in more matured phases (17 to 18 days). Invading inflammatory cells were TNF-alpha immunopositive beginning on the third day. Besides, vascular wall structures showed immunoreactivity sporadically. TNF-alpha levels were mostly nondetectable in peripheral blood. TUNEL labeling and TNF-alpha staining overlapped, although not completely, during the first days. CONCLUSIONS: The data support the hypothesis that TNF-alpha may be involved both in the acute propagation of inflammatory processes and cell death and possibly in the more delayed reconstitutive processes of human ischemic stroke. PMID- 11486102 TI - Matrix metalloproteinase expression after human cardioembolic stroke: temporal profile and relation to neurological impairment. AB - BACKGROUND AND PURPOSE: Uncontrolled expression of matrix metalloproteinases (MMPs) can result in tissue injury and inflammation. In animal models of cerebral ischemia, the expression of MMP-2 and MMP-9 was significantly increased. However, their role in human stroke in vivo remains unknown. Therefore, we sought to determine the temporal profile of MMP expression in patients with acute ischemic stroke and to investigate its relationship to stroke severity, location of arterial occlusion, and total infarct volume. METHODS: Serial MMP-2 and MMP-9 determinations were made in 39 patients with cardioembolic strokes that involved the middle cerebral artery territory by means of enzyme-linked immunosorbent assay. Blood samples, transcranial Doppler recordings, and National Institutes of Health Stroke Scale (NIHSS) scores were obtained at baseline and at 12, 24, and 48 hours after stroke onset. Infarct volume was measured with CT scanning at 48 hours. RESULTS: No correlation was found between MMP-2 and NIHSS score at any time point, although a close relation appeared between mean MMP-9 and final NIHSS score (r=0.486, P=0.002). MMP-9 value was the only factor associated with the final NIHSS score in the multiple logistic regression model (OR 4.54, 95% CI 1.5 to 13.75). A cut-point of MMP-9 142.18 ng/mL had a positive predictive value of 94.4% to assess a patient's NIHSS (<8 or >/=8) by the end of the study. Final MMP 2 and MMP-9 levels were significantly lower when recanalization occurred (528+/ 144.3 versus 681.4+/-239.2 ng/mL, P=0.031 for MMP-2; 110.2+/-100.9 versus 244.8+/ 130 ng/mL, P=0.004 for MMP-9). A positive correlation was found between mean MMP 9 and infarct volume (r=0.385, P=0.022). CONCLUSIONS: MMPs are involved in the acute phase of human ischemic stroke. MMP-9 levels are associated with neurological deficit, middle cerebral artery occlusion, and infarct volume. PMID- 11486103 TI - Phenotype of a homozygous CADASIL patient in comparison to 9 age-matched heterozygous patients with the same R133C Notch3 mutation. AB - BACKGROUND AND PURPOSE: CADASIL is an autosomal dominant arteriopathy, characterized by multiple brain infarcts, cognitive decline, and finally dementia, which is caused by mutations in Notch3 gene encoding a Notch3 receptor protein. We describe the clinical, neuropsychological, imaging, genetic, and skin biopsy findings in a CADASIL patient homozygous for the C475T mutation resulting in R133C amino acid substitution, in comparison to 9 age-matched heterozygous patients with the same mutation. METHODS: The patients were examined clinically and neuropsychologically and with MRI and positron emission tomography for assessment of cerebral blood flow. The gene defect was analyzed by sequencing the products of polymerase chain reaction of exons 3 and 4 of the Notch3 gene. Dermal arteries were analyzed electron microscopically. RESULTS: The homozygous patient had his first-ever stroke at age 28 years. This is markedly earlier than the average, but the patient's heterozygous son had his first transient ischemic attack-like episode at the same age and another heterozygous patient had his first-ever stroke when only 2 years older. He was neuropsychologically more severely deteriorated than all but 1 of the heterozygous patients. These 2 patients had the most severe (confluent grade D) white matter MRI changes. Positron emission tomography showed markedly reduced cerebral blood flow. Skin biopsy revealed profuse deposits of granular osmiophilic material. The progression of disease in the homozygous case was, however, slower than in the most severely affected heterozygous patient. CONCLUSIONS: Our homozygous patient's phenotype is within the clinical spectrum of CADASIL, although at its severe end. Thus, CADASIL may follow the classic definition of a dominant disease, according to which the heterozygous and homozygous patients are clinically indistinguishable. PMID- 11486104 TI - Difference in carotid artery wall structure between Swedish and French men at low and high coronary risk. AB - BACKGROUND AND PURPOSE: We attempted to detect a group-specific north-south difference in carotid artery intima-media thickness (IMT), a marker of subsequent cardiovascular complication, by means of a case (high risk)-control (low risk) study in French and Swedish men. METHODS: The selection of high-risk and low-risk subjects was performed within the lower and upper percentiles of the Framingham risk distribution of 2 samples of 1000 white, male auto workers (45 to 50 years of age) in France (Renault) and Sweden (Volvo). In total, 299 men at low risk (79 French, 76 Swedish) and high risk (61 French, 83 Swedish), free from sustained hypertension, definite hypercholesterolemia, and cardiovascular disease, were included. Both common carotid arteries, by ultrasonography and central off-line computerized analysis, provided measurements of far wall media thickness, lumen diameter, and cross-sectional area IMT (CSA-IMT). RESULTS: As compared with low risk status, high-risk status was associated with higher IMT (P<0.001), diameter (P<0.01), and CSA-IMT (P<0.001) in French men and higher CSA-IMT (P<0.05) in Swedish men. IMT, diameter, and CSA-IMT were higher in Swedish than in French men in the low-risk group (P<0.001) and in the high-risk group (P<0.01, P<0.001, P<0.001). The multivariate analysis of the whole population showed that IMT, diameter, and CSA-IMT were associated with risk status (P<0.01, P<0.01, P<0.001) and geographic status (P<0.001). CONCLUSIONS: These findings show that the geographic status influences carotid artery structure independent of traditional cardiovascular risk factors and that this may affect the mortality and morbidity gradient between Northern and Southern Europe. PMID- 11486105 TI - Significance of earlier carotid atherosclerosis for stroke subtypes. AB - BACKGROUND AND PURPOSE: In addition to advanced stenosis, earlier stages of carotid atherosclerosis are associated with the risk for stroke. However, the significance has not been established for specific stroke subtypes. This study examines the association of earlier carotid atherosclerosis with stroke subtypes. METHODS: The subjects comprised 1059 patients (mean+/-SD age, 62+/-11 years) with <60% carotid stenosis. With the use of ultrasound, carotid atherosclerosis was evaluated by the plaque score, as defined by the sum of all plaque heights in bilateral carotid arteries. On the basis of neurological signs and symptoms, medical history, and brain MRI, we diagnosed stroke and its subtypes as follows: no stroke (n=738), atherothrombotic infarction (AI) (n=56), lacunar infarction (LI) (n=117), cardioembolic infarction (n=65), cerebral hemorrhage (n=26), and other or unclassified stroke (n=57). RESULTS: The plaque score was higher in AI (10.5+/-5.9) and LI (6.0+/-5.1) groups than in the no-stroke group (4.3+/-4.9) (both P<0.05), although it was similar between other stroke groups and the no stroke group. Each 1 SD greater plaque score was associated with 2.5-fold (95% CI, 2.0 to 3.2) higher risk for AI and 1.4-fold (95% CI, 1.2 to 1.7) higher risk for LI compared with the no-stroke group. When we adjusted for cardiovascular risk factors, plaque score remained significantly associated with AI but not with LI. By receiver operating characteristic curve analyses, the receiver operating characteristic area for AI (0.81 to 0.86) was greater than that for LI (0.62 to 0.67) when we used plaque score either alone or in combination with cardiovascular risk factors. CONCLUSIONS: Although evaluation of carotid atherosclerosis may aid in the risk assessment for AI and LI, the benefit appears to be greater for AI. PMID- 11486106 TI - Prothrombotic disorders in children with moyamoya syndrome. AB - BACKGROUND AND PURPOSE: Moyamoya syndrome is an uncommon chronic occlusive cerebrovascular disease in children. The origin of moyamoya syndrome remains undetermined. The role of the prothrombotic disorders contributing to its pathogenesis has not been completely elucidated. The purpose of this study was to determine the frequency of prothrombotic disorders in a pediatric population with moyamoya syndrome. METHODS: From May 1992 to April 2000, a prospective study of 10 consecutive children with moyamoya syndrome was carried out at a single center. Evaluation included the following assays: protein C, protein S, antithrombin, plasminogen, activated protein C resistance, factor V Leiden, and prothrombin gene mutations. Lupus anticoagulant, anticardiolipin antibodies, and anti-beta(2)-glycoprotein I antibodies assays were also performed. The clinical characteristics, underlying diseases, family history of thrombosis, radiological findings, treatment, and outcome were also recorded. RESULTS: In our series, prothrombotic disorders were detected in 4 patients (40%). Inherited protein S deficiency was found in 1 patient; lupus anticoagulant and anticardiolipin antibodies were detected in the remaining 3 patients. One presented persistent lupus anticoagulant for 2.7 years until his death. In the case of the other 2 patients, 1 has maintained lupus anticoagulant for 9 months, whereas the other has kept anticardiolipin/anti-beta(2)-glycoprotein I antibodies for 10 months. CONCLUSIONS: We report the hemostatic data of the largest prospective pediatric study carried out at a single center in the western hemisphere. In 4 patients (40%), a prothrombotic disorder was detected. It is tempting to speculate that these hemostatic abnormalities may contribute to the pathogenesis of moyamoya syndrome in some of our patients. PMID- 11486107 TI - Inherited thrombophilia in ischemic stroke and its pathogenic subtypes. AB - BACKGROUND AND PURPOSE: One or more of the inherited thrombophilias may be causal risk factor for a proportion of ischemic strokes, but few studies have addressed this association or the association between thrombophilia and pathogenic subtypes of stroke. METHODS: We conducted a case-control study of 219 hospital cases with a first-ever ischemic stroke and 205 randomly selected community control subjects stratified by age, sex, and postal code. With the use of established criteria, cases of stroke were classified by pathogenic subtype in a blinded fashion. The prevalence of conventional vascular risk factors; fasting plasma levels of protein C, protein S, antithrombin III; and genetic tests for the factor V Leiden and the prothrombin 20210A mutation were determined in cases and control subjects. RESULTS: The prevalence of any thrombophilia was 14.7% (95% CI, 9.9% to 19.5%) among cases and 11.7% (95% CI, 7.4% to 17.0%) among control subjects (OR, 1.3; 95% CI, 0.7% to 2.3%). The prevalence of individual thrombophilias among cases ranged from 0.9% (95% CI, 0.1% to 3.4%) for protein S deficiency to 5.2% (95% CI, 0.3% to 9.1%) for antithrombin III deficiency; among control subjects, the prevalence ranged from 1.0% (95% CI, 0.1% to 3.6%) for protein S deficiency to 4.1% (95% CI, 0.2% to 7.8%) for antithrombin III deficiency. There were no significant differences in the prevalence of thrombophilia between cases and control subjects or between pathogenic subtypes of ischemic stroke. CONCLUSIONS: One in 7 patients with first-ever acute ischemic stroke will test positive for one of the inherited thrombophilias, but the relation is likely to be coincidental rather than causal in almost all cases, irrespective of the pathogenic subtype of the ischemic stroke. These results suggest that routine testing for thrombophilia in most patients with acute ischemic stroke may be unnecessary. Whether the thrombophilias may still be important in younger patients with ischemic stroke or in predicting complications (eg, venous thrombosis) and stroke outcome remains uncertain. PMID- 11486108 TI - Development of a novel, weighted, quantifiable stroke scale: Japan stroke scale. AB - BACKGROUND AND PURPOSE: Several stroke scales are available for estimation of the severity of stroke, but none of them provides information regarding the relative weights of the observed variables. To define an integrated severity of stroke, we developed a quantifiable stroke scale with weighted variables that apply conjoint analysis to calculate the relative weight of each item. METHODS: We selected 10 variables (consciousness, language, neglect, hemianopsia, gaze, pupillary abnormality, facial palsy, plantar reflex, sensation, and weakness) based on the multivariate analysis of the Keio Stroke Patient Database Battery. The variables were categorized and evaluated for their distribution and sensitivity. The categorizations were then modified and rechecked. The procedure was repeated until the appropriate categorization was obtained from 198 patients. A temporary stroke scale without weight was then formulated, and the reliability of the scale was examined and revised with 80 new stroke patients. As a next step, 150 neurologists were asked to rank a set of 27 virtual patients, each with a different combination of variables, according to severity. From these rankings, conjoint analysis was used to derive utility scores (weights) for each factor level. RESULTS: The relative weights of each of the factors were as follows: consciousness 49.8%, language 9.9%, weakness of lower extremity 7.3%, pupillary abnormality 6.8%, gaze palsy 5.6%, weakness of arm 4.3%, weakness of hand 3.7%, neglect 3.7%, facial palsy 2.4%, plantar reflex 2.2%, hemianopsia 2.2%, and sensory impairment 2.1%. The total score for a patient could be calculated from the sum of the scores for each of the variables ranging from -0.38 to 27.86. Scoring of 100 patients with acute stroke was carried out, and the changes in scores were followed for validation. Longitudinal clinical monitoring of the patients correlated well with the scores in each patient. The interrater and intrarater reliabilities of the scale were excellent (weighted kappa 0.83; Cronbach's alpha 0.998). CONCLUSIONS: The Japan Stroke Scale is a parametric stroke scale that provides a quantitative measure of the severity of stroke. Each of the variables of the scale has a relative weight according to the severity of stroke. Reliability and responsiveness were proved to be excellent. The present data revealed a potentiality for the Japan Stroke Scale to be a universally accepted and reliable standardized system from the clinimetrical point of view. PMID- 11486109 TI - Effect of hemiparetic stroke on pulse oximetry readings on the affected side. AB - BACKGROUND AND PURPOSE: Hypoxia is common after stroke, and monitoring by pulse oximetry is suggested in the acute phase. Physical changes on the affected side or intravenous infusions may affect oximeter readings. This study was designed to test whether pulse oximetry recordings are the same on the affected and nonaffected sides in stroke patients. METHODS: Oxygen saturation (SpO(2)) and heart rate (HR) were assessed simultaneously in the left and right hands in patients with hemiparetic stroke over a 3-hour period with 2 Minolta Pulsox-3i oximeters attached to the index fingers. RESULTS: Fifteen patients (53% men; 67% left hemiparesis; mean age, 73 years [SD, 7.5 years]) were recruited. HR and SpO(2) (12 measurements per minute) were monitored. The maximum difference between simultaneous left and right arm readings was 2% SpO(2). HR fluctuated more, but no affected/nonaffected side pattern was seen. Means for each patient of HR and SpO(2) for the affected and nonaffected sides were compared by t tests. Mean SpO(2) was 96% (SD, 1%) on both sides. Mean HR was 81 bpm (SD, 11 bpm) on the affected side and 80 bpm (SD, 10 bpm) on the nonaffected side. There was no significant difference between the 2 sides for either parameter (n=15; P=0.86 for SpO(2) and P=0.91 for HR). CONCLUSIONS: Oximeters can be attached to either the affected or nonaffected side in hemiparetic stroke. PMID- 11486110 TI - Cerebrovascular reserve in patients with carotid occlusive disease assessed by stable xenon-enhanced ct cerebral blood flow and transcranial Doppler. AB - BACKGROUND AND PURPOSE: Cerebrovascular reserve (CVR) by both transcranial Doppler ultrasonography (TCD) and quantitative cerebral blood flow (CBF) can identify subgroups of patients at increased risk for stroke. A direct comparison of CVR measurements obtained with both technologies in patients with cerebrovascular occlusive disease is lacking. METHODS: CVRs before and after acetazolamide administration (1 g IV) were measured by TCD insonation of the middle cerebral artery (MCA) and CBF obtained with stable xenon CT (Xe/CT) in 38 patients with carotid occlusive disease. Sensitivity/specificity calculations were based on 2 Xe/CT MCA values: an average over 4 levels and the level with the lowest percent change in CBF. Compromised CVR was defined as no reactivity or a decrease in reactivity. RESULTS: Using the analysis of the systolic TCD, we found that velocity changes compared with the average Xe/CT MCA CVR showed a sensitivity of 33%, specificity of 90.6%, positive predictive value of 54.5%, and negative predictive value of 80%. The sensitivity of TCD compared with the lowest Xe/CT CBF CVR was 35.5%, specificity and positive predictive values were 100%, and negative predictive value was 66.7%. The index of validity was between 72% and 76%. CONCLUSIONS: TCD is much less sensitive than Xe/CT CBF in identifying patients with compromised CVR. This may be a result of the inability of TCD to identify patients with compromised reserves when their MCA blood flow comes from collateral sources. The lack of correlation between TCD and Xe/CT CBF for identifying patients with compromised CVR should be considered when stroke risk assessments are made by TCD. PMID- 11486111 TI - Vasospasm after subarachnoid hemorrhage: interest in diffusion-weighted MR imaging. AB - BACKGROUND AND PURPOSE: Vasospasm secondary to subarachnoid hemorrhage (SAH) is responsible for severe ischemic complications. Although effective, angioplasty must be performed at a very early stage to produce any clinical recovery. Diagnostic investigations to assess arterial narrowing (transcranial Doppler, angiography) or cerebral perfusion (xenon CT, single-photon emission CT) do not provide evidence of the extent of parenchymal ischemia. In stroke, diffusion weighted MR imaging (DWI) appears to be the most sensitive procedure to detect cerebral ischemia. We studied asymptomatic vasospasm in patients with aneurysmal SAH to assess whether DWI provides predictive markers of silent ischemic lesions and/or progression toward symptomatic ischemia. METHODS: Seven asymptomatic vasospasm patients (average blood velocity rates >120 cm/s), 3 patients with symptomatic vasospasm, and 4 patients with SAH but without vasospasm were studied at regular intervals by DWI, and their apparent diffusion coefficients (ADCs) were calculated. RESULTS: All patients with vasospasm including those without symptoms presented abnormalities on DWI with a reduction of the ADC prevalently in the white matter. No such abnormalities were observed in patients without vasospasm. The abnormalities on DWI resolved completely in 4 of the 7 patients, with no parenchymal lesion. Resolution was partial in 3 patients whose white matter still presented residual round, focal ischemic lesions. CONCLUSIONS: Being able to correlate abnormalities on DWI with parenchymal involvement in asymptomatic patients would be of considerable clinical significance. It is hoped that larger studies will be undertaken to determine whether the ADC has a reversibility threshold, because this would facilitate patient management. PMID- 11486112 TI - High-resolution turbo magnetic resonance angiography for diagnosis of Moyamoya disease. AB - BACKGROUND AND PURPOSE: High-resolution turbo MR angiography with zero-filling interpolation (ZFI) technique is a new vascular imaging method with reduced scan time. The purpose of the present study was to evaluate high-resolution turbo MR angiography for the diagnosis and assessment of moyamoya disease. METHODS: Forty six patients suspected of having moyamoya disease were examined with high resolution turbo MR angiography with the ZFI technique, MRI, and conventional angiography. Moyamoya disease was diagnosed in 42 of these patients. Blind, separate interpretation of the images was performed. RESULTS: High-resolution turbo MR angiography and MRI accurately evaluated 349 (95%) and 325 (88%) of 368 arteries, respectively, but the degree of stenosis was overestimated in the other arteries. MR angiography and MRI depicted basal cerebral moyamoya vessels in 82 (98%) and 82 (98%) of 84 hemispheres, respectively. MR angiography also depicted leptomeningeal and transdural collateral vessels in 51 (100%) of 51 hemispheres and in 38 (88%) of 43 hemispheres, respectively. The sensitivity and specificity of high-resolution turbo MR angiography for the diagnosis of moyamoya disease were 98% and 100%, respectively. CONCLUSIONS: High-resolution turbo MR angiography in reduced scan time is highly accurate in the assessment of both steno-occlusive lesions and collateral vessels in moyamoya disease, thus providing a highly accurate (98%) diagnosis and assessment of moyamoya disease. PMID- 11486113 TI - Treatment with tissue plasminogen activator and inpatient mortality rates for patients with ischemic stroke treated in community hospitals. AB - BACKGROUND AND PURPOSE: Most analyses of intravenous tissue plasminogen activator (IV tPA) use for acute stroke in routine practice have been limited by sample size and generally restricted to patients treated in large academic medical facilities. In the present study, we sought to estimate among community hospitals the use of IV tPA and to identify factors associated with the use of IV tPA and inpatient mortality. METHODS: We evaluated a retrospective cohort of 23 058 patients with ischemic stroke from 137 community hospitals. RESULTS: Three hundred sixty-two (1.6%) patients were treated with IV tPA, and 9.9% of those patients died during the hospitalization period. In 35.0% of the hospitals, no patients were treated with IV tPA, whereas 14.6% of hospitals treated approximately 3.0% with IV tPA. After control for multiple factors, younger patients, more severely ill patients (OR 2.02, 95% CI 1.36 to 3.01), and patients treated in rural hospitals (OR 1.80, 95% CI 0.99 to 3.26) were more likely to receive IV tPA, whereas black patients were less likely (OR 0.54, 95% CI 0.31 to 0.95). There also was a trend showing that women were less likely to receive IV tPA (OR 0.84, 95% CI 0.69 to 1.03). Factors associated with an increased odds of inpatient mortality included receipt of IV tPA among men (OR 2.81, 95% CI 1.72 to 4.58) and increased age. Black patients were 27% less likely to die during hospitalization (95% CI 0.60 to 0.90). CONCLUSIONS: In this large, retrospective evaluation of community hospital practice, the use IV tPA and inpatient mortality rates among IV tPA-treated patients were consistent with those of other studies. The likelihood of receiving IV tPA varies by race, age, disease severity, and possibly gender. These factors may influence mortality rates. PMID- 11486114 TI - Open-label dose-titration safety and efficacy study of tizanidine hydrochloride in the treatment of spasticity associated with chronic stroke. AB - BACKGROUND AND PURPOSE: Spasticity is a frequently observed motor impairment that develops after stroke; it can cause pain and disability in those affected. The objective of the present study was to evaluate the safety and efficacy of tizanidine, a centrally acting alpha(2)-adrenergic agonist, in the treatment of stroke-related spasticity. METHODS: Forty-seven patients, who were a minimum of 6 months poststroke and had significant spasticity, were studied at 4 centers. Tizanidine was administered in an open-label manner for 16 weeks, beginning at 2 mg/d and slowly titrated to a maximum of 36 mg/d. The Modified Ashworth Scale, muscle strength testing, functional assessments, and Pain and Functional Spasticity Questionnaires were administered at baseline and at 4, 8, 16, and 18 weeks (after 1 week off tizanidine). RESULTS: Spasticity was significantly improved between baseline and week 16, with a decrease in total upper extremity Modified Ashworth Scale score of 2.80+/-0.47 (P<0.0001). No decline in strength was noted. Treatment with tizanidine resulted in a significant improvement in pain intensity (P=0.0375), quality of life (P=0.0001), and physician assessment of disability (P=0.0001). The most frequent side effects were somnolence (62%) and dizziness (32%). No serious adverse events were considered to be drug related. Ten of 47 patients (21%) were able to reach the maximum daily dosage of 36 mg. CONCLUSIONS: Overall, the data suggest that tizanidine is safe and efficacious in the treatment of stroke-related spasticity, preserving muscle strength while reducing muscle tone and painful spasms in affected patients. PMID- 11486115 TI - Cooling for acute ischemic brain damage (cool aid): an open pilot study of induced hypothermia in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Hypothermia is effective in improving outcome in experimental models of brain infarction. We studied the feasibility and safety of hypothermia in patients with acute ischemic stroke treated with thrombolysis. METHODS: An open study design was used. All patients presented with major ischemic stroke (National Institutes of Health Stroke Scale [NIHSS] score >15) within 6 hours of onset. After informed consent, patients with a persistent NIHSS score of >8 were treated with hypothermia to 32+/-1 degrees C for 12 to 72 hours depending on vessel patency. All patients were monitored in the neurocritical care unit for complications. A modified Rankin Scale was measured at 90 days and compared with concurrent controls. RESULTS: Ten patients with a mean age of 71.1+/-14.3 years and an NIHSS score of 19.8+/-3.3 were treated with hypothermia. Nine patients served as concurrent controls. The mean time from symptom onset to thrombolysis was 3.1+/-1.4 hours and from symptom onset to initiation of hypothermia was 6.2+/-1.3 hours. The mean duration of hypothermia was 47.4+/-20.4 hours. Target temperature was achieved in 3.5+/-1.5 hours. Noncritical complications in hypothermia patients included bradycardia (n=5), ventricular ectopy (n=3), hypotension (n=3), melena (n=2), fever after rewarming (n=3), and infections (n=4). Four patients with chronic atrial fibrillation developed rapid ventricular rate, which was noncritical in 2 and critical in 2 patients. Three patients had myocardial infarctions without sequelae. There were 3 deaths in patients undergoing hypothermia. The mean modified Rankin Scale score at 3 months in hypothermia patients was 3.1+/-2.3. CONCLUSION: Induced hypothermia appears feasible and safe in patients with acute ischemic stroke even after thrombolysis. Refinements of the cooling process, optimal target temperature, duration of therapy, and, most important, clinical efficacy, require further study. PMID- 11486116 TI - Endovascular treatment of symptomatic carotid stenosis using stent placement: long-term follow-up of patients with a balanced surgical risk/benefit ratio. AB - BACKGROUND AND PURPOSE: Carotid endarterectomy (CEA) is not necessarily beneficial in all patients with symptomatic high-grade (>/=70%) internal carotid artery (ICA) stenosis. Independent risk factors modulate both the individual stroke risk under medical treatment and the combined stroke and death risk after CEA. Endovascular stenting of symptomatic ICA stenosis may be an alternative to CEA in patients with a balanced surgical risk/benefit ratio. METHODS: We included 43 patients (71% men; median age, 67 years) with a recently symptomatic ICA stenosis with >/=70% luminal narrowing in whom the individual sum of medical and surgical risk factors suggested a balanced surgical risk/benefit ratio (risk modeling appraisal derived from the European Carotid Surgery Trial). After stenting of the stenosed ICA with distal balloon protection, the mean+/-SD follow up, including clinical and ultrasonographic examinations, was 20+/-11.8 months, with a median number of examinations of 5 per patient. RESULTS: Recanalization of ICA stenoses was technically successful in 40 of 43 procedures (93%). Within the 30-day postinterventional period 1 death occurred (2.5%), and the combined stroke and death rate within follow-up was 5%. Except for 1 asymptomatic ICA occlusion, no restenosis >/=70% occurred during follow-up. CONCLUSIONS: ICA stenting in symptomatic patients with a balanced surgical risk/benefit ratio is technically feasible, with a low periprocedural risk of stroke or death. Furthermore, the risk of future stroke and rate of significant restenosis during long-term follow up appears to be low, suggesting that ICA stenting may be useful in carotid revascularization and stroke prevention. PMID- 11486117 TI - Nimodipine increases fibrinolytic activity in patients with aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: The purpose of this study was first, to investigate which factor in the fibrinolytic cascade is responsible for the recently observed increase of fibrinolytic activity in patients with aneurysmal subarachnoid hemorrhage (SAH), and second, the cause of this increase. METHODS: Fibrinolytic activity and the main regulators of endogenous fibrinolytic activity, tissue plasminogen activator, and plasminogen activator inhibitor 1 (PAI-1) were measured in patients treated with and without nimodipine. RESULTS: In patients with aneurysmal SAH, fibrinolytic activity significantly increases from 2.7 IU/mL on admission to 4.2 IU/mL in week 3 (P<0.01, paired-sample t test), caused by a 1.6-fold decrease in plasma levels of PAI-1. The results also show that increased fibrinolytic activity is seen only in patients treated with nimodipine and that plasminogen activity and PAI-1 returned to baseline levels after treatment with nimodipine had been discontinued. CONCLUSIONS: The mechanism of increased fibrinolytic activity in patients with aneurysmal SAH is a decrease in the level of PAI-1, which is most likely caused by treatment with nimodipine. PMID- 11486118 TI - Predictive value of neurochemical monitoring in large middle cerebral artery infarction. AB - BACKGROUND AND PURPOSE: Space-occupying brain edema is a life-threatening complication in patients with large hemispheric stroke. Early identification of patients at risk is necessary to decide on invasive therapies such as decompressive hemicraniectomy or hypothermia. To assess potential predictors of malignant brain edema by measurement of intracranial pressure (ICP) and microdialysis in patients with large hemispheric stroke and different clinical course. METHODS: In an ongoing prospective clinical study, an ICP and microdialysis probe were placed into the parenchyma of the ipsilateral frontal lobe of 10 patients. Extracellular concentrations of glutamate, lactate, pyruvate, and glycerol were measured continuously. Repeated cranial CT scans were scrutinized for size of infarction and presence of mass effect. RESULTS: The dynamics of the different substances varied in accordance with the clinical course, size of infarction, and local brain edema: Increase in ICP and in glutamate concentration and lactate-pyruvate ratio was followed by massive edema and large infarcts; generally low and stable ICP and substrate concentrations were found in patients without progressive space-occupying infarcts. CONCLUSIONS: In patients with large hemispheric infarction, bedside monitoring with microdialysis is feasible and might be helpful together with ICP recording to follow the development of malignant brain edema. PMID- 11486119 TI - Vasospasm in monkeys resolves because of loss of and encasement of subarachnoid blood clot. AB - BACKGROUND AND PURPOSE: We studied in monkeys why vasospasm resolves after subarachnoid hemorrhage (SAH). METHODS: Monkeys underwent angiography and right (n=17) or bilateral (n=8) SAH. Animals with bilateral SAH underwent angiography 1, 3, 5, and 7 days later. Animals with right SAH underwent angiography 7 days later. The clot was then not removed (n=5), removed and replaced with fresh clot (n=7), or removed and not replaced (n=5). At the same time on day 7, the removed clot (n=12) or fresh clot (n=5) was placed on the left side. Angiography was repeated every 2 days until day 14. RESULTS: SAH caused significant vasospasm on day 7 that resolved by day 14. Removal of clot on day 7 resulted in more rapid resolution of vasospasm. Placement of fresh clot onto arteries that had already been exposed to clot for 7 days produced vasospasm that persisted without resolving for an additional 7 days. Placement of 7-day-old clot from the right onto previously unexposed left arteries or of clot from blood removed from an animal 7 days after SAH caused significantly more rapid onset of vasospasm compared with de novo vasospasm. Microscopic examination of the clots showed they were surrounded by macrophages 7 days after SAH. Arterial compliance and contractility were reduced in relation to duration of the exposure of arteries to clot. CONCLUSIONS: Vasospasm resolves because of loss of subarachnoid blood clot. We hypothesize that reduced spasmogen release from the clot contributes to resolution of vasospasm. There was no response in the cerebral arteries that rendered them less responsive to the subarachnoid clot. PMID- 11486120 TI - Effect of trunk restraint on the recovery of reaching movements in hemiparetic patients. AB - BACKGROUND AND PURPOSE: Reaching movements made with the affected arm in hemiparetic patients are often accompanied by compensatory trunk or shoulder girdle movements, which extend the reach of the arm. We investigated the effects of the suppression of these compensatory movements on reaching ability in hemiparetic individuals. METHODS: Eleven healthy and 11 hemiparetic individuals participated. Three-dimensional kinematic analysis was used to quantify reaches made to a close and a distant target (near the limit of arm's length). Unrestrained reaches were compared with those in which shoulder girdle and trunk movements were restrained by a harness. RESULTS: During unrestrained reaching, abnormal trunk recruitment and limitations in elbow and shoulder movements were correlated with the degree of clinical stroke severity (r=-0.91 to -0.96) in hemiparetic patients. During trunk restraint, ranges of elbow and shoulder joint movement increased in both groups. In addition, elbow and shoulder interjoint coordination improved. This was caused by increases in the range of joint motion as well as by a better dynamic temporal relation between joints. CONCLUSIONS: Trunk restraint allowed patients with hemiparetic stroke to make use of arm joint ranges that are present but not normally recruited during unrestrained arm reaching tasks. Thus, the underlying "normal" patterns of movement coordination may not be entirely lost after stroke. Appropriate treatments, such as trunk restraint, may be effective in uncovering latent movement patterns to maximize arm recovery in hemiparetic patients. PMID- 11486121 TI - Compliance with secondary prevention of ischemic stroke: a prospective evaluation. AB - BACKGROUND AND PURPOSE: Compliance with pharmacological therapy is essential for the efficiency of secondary prevention of ischemic stroke. Few data exist regarding patient compliance with antithrombotic and risk factor treatment outside of controlled clinical trials. The aim of the present study was to assess the rate of and predictors for compliance with secondary stroke prevention 1 year after cerebral ischemia and to identify reasons for noncompliance. METHODS: Patients with a diagnosis of ischemic stroke or TIA and antithrombotic discharge medication were prospectively recruited. At 1 year, the proportion of patients compliant with antithrombotic treatment and with medication for risk factors (eg, hypertension, diabetes, hyperlipidemia) was evaluated through structured telephone interviews. In addition, the reasons for nontreatment with antithrombotic and risk factor medication were determined. Independent predictors for compliance were analyzed by logistic regression analyses. RESULTS: Of 588 consecutive patients admitted to our stroke unit, 470 had a discharge diagnosis of cerebral ischemia (TIA 26.2%, cerebral infarct 73.8%) and recommendations for antithrombotic therapy. At 1 year, 63 patients (13.4%) had died and 21 (4.5%) were lost to follow-up, thus, 386 could finally be evaluated. Of the patients, 87.6% were still on antithrombotic medication, and 70.2% were treated with the same agent prescribed on discharge. Of the patients with hypertension, diabetes, and hyperlipidemia, 90.8%, 84.9%, and 70.2% were still treated for their respective risk factors. Logistic regression analyses revealed age (OR 1.03, 95% CI 1.00 to 1.06), stroke severity on admission (OR 1.09, 95% CI 1.00 to 1.20), and cardioembolic cause (OR 4.13, 95% CI 1.23 to 13.83) as independent predictors of compliance. CONCLUSIONS: Compliance with secondary prevention in patients with ischemic stroke is rather good in the setting of our study. Higher age, a more severe neurological deficit on admission, and cardioembolic stroke cause are associated with better long-term compliance. Knowledge of these determinants may help to further improve the quality of stroke prevention. PMID- 11486122 TI - Neurogenesis by progenitor cells in the ischemic adult rat hippocampus. AB - BACKGROUND AND PURPOSE: Recently, there has been great interest in adult neurogenesis. We investigated whether transient forebrain ischemia could influence the proliferation of neuronal progenitor in the subgranular zone (SGZ) of the rat hippocampus and whether aging could influence the neurogenesis after ischemia. METHODS: Male Wistar rats were subjected to 4-vessel occlusion model. We used a bromodeoxyuridine (BrdU) labeling method to identify the postproliferation cells and double-immunostaining with confocal microscopy to determine the cell phenotype. RESULTS: The number of BrdU-positive cells in the SGZ increased approximately 5.7-fold 8 days after ischemia, compared with the control. BrdU-positive cells formed clusters, which suggested that these cells had divided from an original progenitor cell, and expressed Musashi1 (Msi1), a marker of neural stem/progenitor cells. Although astrocytes also expressed Msi1 in the adult brain, Msi1-positive cells that formed clusters in the SGZ did not express glial fibrillary acidic protein, an astrocyte marker. About 70% of all BrdU-positive cells in the SGZ represented the neuronal phenotype 4 weeks after the BrdU injection. Although proliferation of progenitor cells was stimulated in both young and older animals, aging accelerated the reduction in newborn cells after ischemia. CONCLUSIONS: Our results indicate that ischemic stress stimulated the proliferation of neuronal progenitor cells in the SGZ of both young and old rats but resulted in increased neurogenesis only in young animals. Our findings will be important in developing therapeutic intervention to enhance endogenous neurogenesis after brain injury. PMID- 11486123 TI - Regional variations in the apparent diffusion coefficient and the intracellular distribution of water in rat brain during acute focal ischemia. AB - BACKGROUND AND PURPOSE: The apparent diffusion coefficient of water (ADC) rapidly drops in ischemic tissue after cerebral artery occlusion. This acute drop is thought to be caused by the loss of extracellular fluid and the gain of intracellular fluid. To test the latter possibility, changes in ADC and the size of several cellular compartments were assessed in 3 regions of rat brain at the end of 90 minutes of focal cerebral ischemia. METHODS: One middle cerebral artery was permanently occluded in 8 Sprague-Dawley rats; sham occlusions were performed in 2 other rats. ADC maps were generated 90 minutes later, and the brains were immediately perfusion fixed. Three regions of interest (ROIs) were defined on the basis of ADC range. Various neuronal, astrocytic, and capillary compartments in each ROI were quantified with light and electron microscopy. RESULTS: At the end of 90 minutes of ischemia, mean ADC was normal in the cortex of sham-operated rats and the contralateral cortex of ischemic rats (ROI-a), 25% lower in the ipsilateral frontoparietal cortex (ROI-b), and 45% lower in the ischemic lateral caudoputamen (ROI-c). At this time, the frequency of swollen astrocytic cell bodies and volume of swollen dendrites and astrocytic processes in neuropil were ROI-a/=1 established stroke risk factor, but only 409 (49.8%) respondents correctly listed >/=1 warning sign. CONCLUSIONS: The level of knowledge in the community of established stroke risk factors, warning signs, and treatment as indicated by this survey suggests that a community-based education program to increase public knowledge of stroke may contribute to reducing the risk of stroke and to increasing the speed of hospital presentation after the onset of stroke. PMID- 11486128 TI - Atrial fibrillation and stroke: more concepts and controversies. PMID- 11486129 TI - Does antibody to the alpha4 integrin inhibit the function of lymphocytes and monocytes? PMID- 11486130 TI - Risk of subarachnoid hemorrhage from a de novo aneurysm. PMID- 11486131 TI - Acupuncture and stroke rehabilitation. PMID- 11486132 TI - More complications of spinal manipulation. PMID- 11486133 TI - Helicobacter pylori and cerebrovascular disease. PMID- 11486134 TI - The volume of lacunes. PMID- 11486135 TI - Best evidence medical education and the perversity of humans as subjects. PMID- 11486136 TI - The effect of examinee and patient ethnicity in clinical-skills assessment with standardized patients. AB - CONTEXT: Ethnicity has been a continuing concern for the valid assessment of clinical performance with standardized patients (SPs). The concern is that examinee ethnicity and SP ethnicity might interact, such that examinees might score higher in encounters with SPs of the same ethnicity. OBJECTIVE: To test for an interaction of examinee ethnicity and SP ethnicity on clinical performance in an SP examination. MAIN OUTCOME MEASURES: History-taking and physical-examination scores and interpersonal-and communication-skills scores, both based on checklists completed by SPs. Poststation scores for answers to case-related questions concerning pathophysiology, diagnosis, test selection, and test interpretation. SETTING AND PARTICIPANTS: Two graduating classes of over 1,000 fourth-year medical students each in the New York City Consortium were tested on the SP assessment administered at The Morchand Center of Mount Sinai School of Medicine. DESIGN AND ANALYSIS: The primary analyses were two-way (2 x 2) analyses, to test the main and interaction effects of examinee ethnicity and SP ethnicity. Effect-size measures (standardized mean differences, d) were computed to provide a sharper picture of the effects. RESULTS: Of the 24 interaction analyses, only three were statistically significant (not significantly more than expected by chance) and the results were mixed: one analysis showed better examinee performance in encounters with SPs of the same ethnic background and the other two showed the opposite. For all 24 interactions, significant or not, the results showed weak effects and no consistent pattern. White examinees scored on average 0.12 standard deviations above black examinees in encounters with white SPs, and 0.11 standard deviations higher in encounters with black SPs. CONCLUSIONS: These initial results are encouraging and should dispel some of the concern about ethnicity in SP assessment, at least about the operation of an examinee-by-SP-ethnicity interaction that would pose a serious threat to the validity of the examination scores. PMID- 11486137 TI - Medication calculation skills of graduating nursing students in Finland. AB - The aim of this study was to describe the basic mathematical proficiency and the medication calculation skills of graduating nursing students in Finland. A further concern was with how students experienced the teaching of medication calculation. We wanted to find out whether these experiences were associated with various background factors and the students' medication calculation skills. In spring 1997 the population of graduating nursing students in Finland numbered around 1280; the figure for the whole year was 2640. A convenience sample of 204 students completed a questionnaire specially developed for this study. The instrument included structured questions, statements and a medication calculation test. The response rate was 88%. Data analysis was based on descriptive statistics. The students found it hard to learn mathematics and medication calculation skills. Those who evaluated their mathematical and medication calculation skills as sufficient successfully solved the problems included in the questionnaire. It was felt that the introductory course on medication calculation was uninteresting and poorly organised. Overall the students' mathematical skills were inadequate. One-fifth of the students failed to pass the medication calculation test. A positive correlation was shown between the student's grade in mathematics (Sixth Form College) and her skills in medication calculation. PMID- 11486138 TI - Interdisciplinary health professional education: a historical review. AB - A historical review was conducted to examine the advances made, nationally and internationally, in interdisciplinary health professional education since the mid 1960s. One hundred and nineteen articles were reviewed and divided by decade into the following subheadings: models, courses, communication/group process issues, and international perspectives. Twenty-seven articles, categorized as models, defined the conceptual field, described curriculum and program development, or provided a framework for evaluation. Thirty-two articles dealt with interdisciplinary courses, focusing on objectives, content areas, or innovative methods. Nine articles contributed content on interdisciplinary communication and group process issues, from which guidelines were abstracted. Fifty-one articles contributed an international perspective, leading to the recognition that interdisciplinary health professional education, practice, and research is a global movement. PMID- 11486139 TI - Philosophical dialogue among peers: a study of manifestations of critical thinking in pre-service teachers. AB - In this paper, we study manifestations of critical thinking in pre-service physical education (PE) teachers when exposed to regular practice of philosophical dialogue among peers. A 12-week research project involving 13 pre service teachers in PE was conducted. Analysis of two (pre- and post-project) transcripts from pre-service teachers' discussions reveals greater use of higher order thinking (HOT) skills in the post-project transcript than in the pre project transcript, and reveals that the nature of their exchanges evolved from monologue to dialogue. The latter transcript reveals some characteristics of critical thinking among participants, which thinking appears to be critical, creative and caring; it also reveals that the participants' thinking was oriented toward social and ethical dimensions. PMID- 11486140 TI - The Liverpool brief assessment system for communication skills in the making of doctors. AB - The teaching of clinical communication skills is gaining importance in medical schools. There is a need to design feasible assessments that are credible to faculty, students and the profession. AIM: To design, and assess the reliability and validity of a new communication skills assessment system (the Liverpool Communication Skills Assessment Scale-LCSAS, and the Global Simulated Patient Rating Scale-GSPRS) for employment in OSCEs. PARTICIPANTS: The first three annual intakes of students to the new medical curriculum at Liverpool Medical School (n = 600). Two further sub samples of 1st year (n = 60) and 2nd year students (n = 80) were included for further validation purposes in separate studies. STATISTICAL ANALYSIS: Intra class correlation coefficients and generalisability coefficients were employed to assess reliability of the LCSAS and GSPRS. Validity was tested by examining predicted relationships using pearson product moment correlation coefficients. RESULTS AND CONCLUSIONS: The LCSAS and GSPRS showed reasonable evidence for their reliability and validity although further work is recommended. PMID- 11486141 TI - What happens to the student? The neglected variable in educational outcome research. AB - Disputes about the superiority of teaching methods often remain unresolved. The essential question we continuously want to answer is: Which teaching methods yield the best knowledge and skills in students? Abundant literature, in medical education and in education in general, on research with educational methods as independent variables and measures of outcome (e.g., test scores) as the dependent variable often point at "no significant difference" or only small differences between methods. Many factors do influence the educational outcome in students and large statistical power (such as meta analysis) should be helpful to eliminate many sources of error. However, one source we cannot tackle this way. That is, students will usually adapt quantity and quality of studying to meet testing requirements. In doing so, they may compensate for teaching quality. Some teaching may generate more effort in students than other teaching. Since test scores reflect primarily student activities, it is their efforts that may bring differences in teaching methods close to equality in test scores. Therefore, knowledge and skills should not be considered the primary outcome of teaching but the outcome of learning activities. If we want to discriminate between teaching methods, we must at least consider what happens to students. PMID- 11486142 TI - [Comparative analysis of Brazil, Mercosul, Great Britain and European Union of food labeling legislation]. AB - OBJECTIVE: To analyze the Brazilian/Mercosul and the British/European Union food labeling legislation. METHODS: The analysis was carried out using a set of 19 questions considered important to describe and compare the different sets of legislation. RESULTS: The results showed that all sets of legislation were very similar. Regarding health claims, the legislation differentiates content from effect claims. However, it is shown to be difficult to do so because both of them express the same message, that a product is good for the health. Concerning nutrition label, the problem is that in all sets of legislation it is only compulsory when a health claim is made. Another problem is that the nutritional description is required to be by weight, while international nutritional recommendations are made as percentage of energy. CONCLUSIONS: Health claims should not be allowed, they are potentially misleading and do not convey more information than nutrition label. Nutrition label should be compulsory, and the macronutrients presented as percentage of energy together with the international nutritional recommendations. PMID- 11486143 TI - A mathematical model for malaria transmission relating global warming and local socioeconomic conditions. AB - OBJECTIVE: Sensitivity analysis was applied to a mathematical model describing malaria transmission relating global warming and local socioeconomic conditions. METHODS: A previous compartment model was proposed to describe the overall transmission of malaria. This model was built up on several parameters and the prevalence of malaria in a community was characterized by the values assigned to them. To assess the control efforts, the model parameters can vary on broad intervals. RESULTS: By performing the sensitivity analysis on equilibrium points, which represent the level of malaria infection in a community, the different possible scenarios are obtained when the parameters are changed. CONCLUSIONS: Depending on malaria risk, the efforts to control its transmission can be guided by a subset of parameters used in the mathematical model. PMID- 11486144 TI - [Larval susceptibility to chemical insecticides of two Aedes aegypti populations]. AB - OBJECTIVE: Insect susceptibility has been one of the most important aspects to be monitored in public health programs for vector control. The purpose of the study is to assess the susceptibility to chemical insecticides of Aedes aegypti larvae in both areas under vector control and no vector control. METHODS: World Health Organization standard bioassays for diagnostic concentration and multiple concentrations were performed in mosquito larvae collected in an area under no vector control (Campinas, SP) and an area under vector chemical control (Campo Grande, MS), in Brazil. RESULTS: Potential resistance to a diagnostic concentration of temephos (DC=0.04 ppm) was registered for an Ae. aegypti larval population collected in Campinas. Multiple concentration tests confirmed the larvae resistance, with 24.5% of them surviving at the 0.0125 ppm concentration. Bioassays with the organophosphate fenitrothion (DC=0.08 ppm) and pyrethroid cypermethrin (DC=0.01 ppm) in the same population revealed their susceptibility to these agents. Bioassays carried out in an Ae. aegypti larval population collected in Campo Grande showed their susceptibility to temephos (DC=0.04 ppm) and cypermethrin (DC=0.01 ppm). LC50 and LC95 for cypermethrin (CE25), cyfluthrin (CE5), betacyfluthrin (SC1.25) and propoxur (CE20) were determined for Ae. aegypti. Using the Rockefeller standard strain values, ratios of resistance were estimated: 2.9, 2.2, 2.4 and 1.3 for LC50 and 3.5, 2.6, 3.9 and 1.3 for LC95, respectively. CONCLUSION: The findings reinforce the need for routinely monitoring pesticide efficacy as a very important step in vector control management programs. PMID- 11486145 TI - [Aedes aegypti (l.) population in an endemic area of dengue in Southeast Brazil]. AB - OBJECTIVE: A Aedes aegypti population from an area of dengue transmission was studied to determine mosquito's frequency, spatial distribution, parity, its ovarian development and midgut contents. METHODS: The study carried out in Sao Jose do Rio Preto country, Sao Paulo, SP, Brazil. Low and intermediate socioeconomic level regions were chosen to the study, which was carried out 1996 and 1997. Using a manual electric aspirator, captures were performed inside and outside the houses. Dissections were conducted to allow classify female mosquitoes according to their physiologic state. RESULTS: One hundred and eighty eight males and 189 females were captured in both regions. A general index of 0.46 Ae. aegypti females per house was determined. Of the total sample, 82.4% of males and 87.3% of females were collected indoors. Greater proportions of females were found in the region of low socioeconomic level, and higher population density. One hundred and forty-eight female mosquitoes were analyzed: 27.0% were nulliparous and 10.1% parous. The rest were classified as Christophers and Mer's (C & M) phases III to V, 28.0% of which had red blood in their midgut. Of the total female population, 87.9% had bloodmeals. CONCLUSIONS: The mosquito species revealed a high degree of endophily. There was a higher proportion of nulliparous mosquitoes, although the majority of the females were classified as C & M's phases III to V. These results highlight both the great number of females who had had bloodmeals, and the lack of gonotrophic concordance. PMID- 11486146 TI - [Aedes albopictus in bromeliads of anthropic environment in Sao Paulo State, Brazil]. AB - OBJECTIVE: Aedes albopictus populations can breed in several kinds of containers, and its presence has also been reported in Bromeliaceae. The purpose of this study was to evaluate the epidemiological importance of the Bromeliaceae plants as potential breeding sites of Aedes albopictus and to document the associated Culicidae entomofauna found in this micro-habitat. METHODS: Collections of Culicidae larvae were carried out fortnightly in aquatic content of bromeliads during 1998 and 1999. Collections took place in urban and periurban areas of Ilhabela island and periurban area of Ilha Comprida island, Brazil. RESULTS: A total of 26,647 Culicidae larvae were collected at both study sites, 14,575 in the urban area and 10,987 in the periurban area of Ilhabela and the remaining 1,085 in the periurban area of Ilha Comprida. There was no statistical difference between the amount of larvae collected in urban and periurban areas of Ilhabela. Regarding the Ae. albopictus, there was found a higher frequency and amount of larvae in the urban area of Ilhabela, followed by the periurban area in the same site, whereas in the periurban area of Ilha Comprida, its presence was considered accidental, since it was reported only in the first three months. CONCLUSIONS: The results show that bromeliads, largely used in landscaping, may contribute to the spread of Ae. albopictus. The presence of this species in domesticated and domiciled bromeliads allows us to suggest that the Culicidae larva is part of the regional fauna and facilitate the contact between humans and etiological agents of the natural ecosystem. PMID- 11486147 TI - Epidemiology of bacterial meningitis among children in Brazil, 1997-1998. AB - OBJECTIVE: To document the incidence and the descriptive epidemiology of bacterial meningitis among individuals under age 20 in a geographically defined region in Brazil during the two-year period immediately preceding the introduction of Haemophilus influenzae type b (Hib) vaccines into the national immunization program of Brazil. METHODS: Population-based epidemiological study of all cases of bacterial meningitis reported among residents of Campinas, Brazil, under age 20 (n=316,570) during the period of 1997-98, using comprehensive surveillance records compiled by the Campinas Health Department from cases reported among hospital inpatients, outpatients, emergency room visits, death certificates, and autopsy reports. RESULTS: The incidence of bacterial meningitis (n=274) was 334.9, 115 and 43.5 cases/10(5) person-years (pys) for residents of Campinas under age 1, 5 and 20, respectively. All cases were hospitalized, with an average length of stay of 12 days. Documented prior antibiotic use was 4.0%. The case-fatality rate of bacterial meningitis in individuals under age 20 was 9% (24/274) with 75% of deaths occurring in children under the age of five. The incidence of Hib meningitis (n=26) was 62.8 and 17 cases/10(5) pys in children age <1 and <5, respectively. CONCLUSIONS: The incidence of Hib meningitis in children under the age of 5 in Campinas during 1997-98 was similar to that reported in the US, Western Europe, and Israel prior to widespread Hib vaccine use in those regions. This study provides a baseline for later studies to evaluate changes in the etiology and incidence of bacterial meningitis in children after introduction of routine Hib vaccination in Brazil. PMID- 11486148 TI - Infant mortality rates according to socioeconomic status in a Brazilian city. AB - OBJECTIVE: Data from municipal databases can be used to plan interventions aimed at reducing inequities in health care. The objective of the study was to determine the distribution of infant mortality according to an urban geoeconomic classification using routinely collected municipal data. METHODS: All live births (total of 42,381) and infant deaths (total of 731) that occurred between 1994 and 1998 in Ribeirao Preto, Brazil, were considered. Four different geoeconomic areas were defined according to the family head's income in each administrative urban zone. RESULTS: The trends for infant mortality rate and its different components, neonatal mortality rate and post-neonatal mortality rate, decreased in Ribeirao Preto from 1994 to 1998 (chi-square for trend, p<0.05). These rates were inversely correlated with the distribution of lower salaries in the geoeconomic areas (less than 5 minimum wages per family head), in particular the post neonatal mortality rate (chi-square for trend, p<0.05). Finally, the poor area showed a steady increase in excess infant mortality. CONCLUSIONS: The results indicate that infant mortality rates are associated with social inequality and can be monitored using municipal databases. The findings also suggest an increase in the impact of social inequality on infant health in Ribeirao Preto, especially in the poor area. The monitoring of health inequalities using municipal databases may be an increasingly more useful tool given the continuous decentralization of health management at the municipal level in Brazil. PMID- 11486149 TI - [Mortality rate associated to traffic accidents and registered motor vehicles]. AB - OBJECTIVE: The mortality rate due to traffic accidents is an information often used while making public health policies. In order to measure traffic violence, a study was carried to analyze the number of death by traffic accidents per registered motor vehicle. METHODS: Based on the number of registered vehicles, population and traffic accident deaths, obtained from the Statistical Yearbook (1999), Demographic Yearbook (1997), Denatran (1999), Ministerio da Saude (2000) and Fundacao IBGE (2000) 61 countries and 51 areas in Brazil were studied. Traffic accident mortality rate was broken down into registered motor vehicles per capita and the number of deaths per motor vehicle. To assess this relationship, the samples (international and Brazilian) were subdivided into three groups according to the number of vehicles per capita. To test the statistical significance of this relationship, log-linear regression was used. RESULTS: The number of deaths per motor vehicles is associated to the number of registered motor vehicles per capita. The statistical relationship is -1.067 for international data and -0.515 for the Brazilian data. CONCLUSIONS: To prevent traffic accidents, there is a need of further studies to explain the high numbers of deaths due to motor vehicles associated with the low rate of registered motor vehicles per capita. PMID- 11486150 TI - [Prevalence and associated factors to rural occupational accidents, Brazil]. AB - OBJECTIVE: To investigate the frequency of rural occupational accidents and its association with potential risk factors. METHODS: The study was carried out in a urban area of Pelotas, RS, Brazil. A cross-sectional study was conducted. A representative sample of rural workers was selected using a multi-stage sampling. From January to April 1996, a total of 258 rural families were visited and all the 580 rural workers identified in those households answered a standardized questionnaire. RESULTS AND CONCLUSIONS: The prevalence of rural occupational accidents was 11%. After allowing for potential confounders, the main occupational hazards, as shown by logistic regression, were lower socioeconomic status (OR=1.81), non-white skin color (OR=3.50), and poor work satisfaction (OR=2.77). PMID- 11486151 TI - [Dental fluorosis in upper permanent incisors of public schoolchildren in Rio de Janeiro, Brazil]. AB - OBJECTIVE: To assess the prevalence and severity of dental fluorosis in children aged 7 to 12. METHODS: The study population comprised 266 children aged 7 to 12 years, enrolled in a public school in the city of Rio de Janeiro, Brazil, selected using a simple random sample method. All examinations were carried out between August and December 1999 by a single trained and calibrated examiner (Kappa = 0.92). After obtaining their parents' consent the children had their upper permanent incisors examined under natural light. Their teeth were previously cleaned and dried with cotton rolls. The differential diagnosis between milder forms of dental fluorosis and nonfluoride enamel opacities was made according to Russel's diagnostic criteria. The severity of fluorosis was assessed by the Thylstrup and Fejerskov Index (TF). RESULTS: The prevalence of dental fluorosis was 7.9% (CI 95%, 5.0-11.8). The severity ranged from 1 to 3 and 77% of the affected teeth had a TF score of 1. CONCLUSION: These results suggest that dental fluorosis does not represent a public health problem in this population. PMID- 11486152 TI - [Prevalence and severity of dental caries among schoolchildren aged six and twelve]. AB - OBJECTIVE: To determine the prevalence and severity of dental caries and to assess treatment needs among schoolchildren aged 6 and 12 in Blumenau, Brazil, and to compare the results found for private and public schools. METHODS: A schoolchildren population of Blumenau, SC, Brazil, was studied. A random sample of 1,473 schoolchildren from Blumenau, Brazil, was obtained. Four strata were studied: 6 and 12 years old schoolchildren attending public and private schools. WHO (1997) criteria for caries diagnosis and treatment needs were used. RESULTS: The prevalence of caries in the primary dentition of 6-years-old children was 60.9% and 34.9% in public and private schools (p<0.0001), respectively. The mean DMF-T for children aged 6 was 2.98 and 1.32 for public and private schools, respectively, (p<0.0001), and 2.42 for the whole sample. The prevalence of caries in the permanent dentition of 12-years-old children was 54.7% and the mean DMF-T was 1.46 for public schools. The response rate for private schools was low and the results could not be analyzed. Forty-two percent and 77.0% of children aged six of public and private schools, respectively, had no treatment needs. For those aged 12, the percentage was 53.0% for public schools. CONCLUSIONS: The prevalence of caries and the treatment needs among schoolchildren in Blumenau was low, but there was a statistically significant higher prevalence and higher severity in public school children aged six. The prevalence of caries and treatment needs at the age of 12 in public schools were also low. PMID- 11486153 TI - [Gingival and periodontal conditions associated with socioeconomic factors]. AB - OBJECTIVE: To assess the prevalence of gingival bleeding, calculus and periodontal pockets among 18-year-old males. In addition, to verify whether these conditions are associated with years of education and family income. METHODS: A cross sectional survey was carried out. A representative sample of 18-year-old males living in Florianopolis, Brazil, was selected from the Brazilian Army applicant list. Clinical and socioeconomic data were collected through dental examinations and interviews, respectively. Socioeconomic data included the subjects and their parents' years of education, and family income. A single dentist carried out the dental examinations looking for gingival bleedings after probing, calculus and shallow and deep periodontal pockets (3.5 mm to 5.5 mm and 5.5 mm and over). The associations between periodontal conditions and socioeconomic variables were analyzed using the qui-square test. RESULTS: A total of 286 people participated in the study. Intra-examiner agreement was high. All kappa values were above 0.7. The prevalence of gingival bleeding, calculus, shallow and deep periodontal pockets were 86%, 50.7%, 7.7% and 0.3% respectively. There were associations between gingival bleeding and all socioeconomic variables (p<0.001). Calculus were associated with the subject's and their father's low educational level (p<0.05) and with their mother's low educational level (p<0.01). Periodontal pockets were associated with their father's low educational level (p<0.05). CONCLUSIONS: The prevalence of periodontal pockets was very low, while gingival bleeding and calculus were high. General periodontal health of the studied population was good. PMID- 11486154 TI - [Generalized anxiety disorder in a population aged 80 years and older]. AB - OBJECTIVE: To describe the prevalence of generalized anxiety disorder (GAD) in a community-dwelling population aged 80 years and older. Also to compare sleep patterns, cognitive function and prevalence of other psychiatric conditions between healthy controls and GAD subjects. METHODS: DSM-IV criteria were used to diagnose GAD. A representative random sample of 77 community-dwelling subjects (35%), aged 80 years or more from the rural southern county of Veranopolis, Brazil, was selected. Sleep patterns were assessed using the Pittsburgh Sleep Quality Index and a 2-week sleep/wake diary. Five neuropsychological tests (Buschke-Fuld Selective Reminding Test, CERAD word list, verbal fluency test and two sub-tests of the Wechsler memory scale) were used for cognitive evaluation. RESULTS: The estimated GAD prevalence was 10.6 percent. GAD was associated with major depression, with a significant greater number of depressive symptoms as measured by the Geriatric Depression Scale and with a significant more frequent occurrence of minor depression. Sleeping patterns and cognition function among GAD subjects were not compromised. The severity of physical diseases was similar in both GAD subjects and healthy controls. GAD was associated with a significantly lower health-related quality of life. CONCLUSIONS: In comparison with previous studies, the prevalence of GAD is higher in the studied population. This disorder is frequently associated with both depressive symptoms and a lower health-related quality of life. PMID- 11486155 TI - Adverse drug reaction reporting. PMID- 11486156 TI - Effect of beta-blockade on the premature ventricular beats/heart rate relation and heart rate variability in patients with coronary heart disease and severe ventricular arrhythmias. AB - We examined the effects of beta-blockers on the associations between heart rate and number of premature ventricular beats (PVBs) and on heart rate variability and myocardial ischemia in patients with coronary heart disease. After 2 weeks of run-in placebo treatment, 18 patients with coronary artery disease were randomized to a 7-day treatment with either propranolol (40 mg) three times a day or placebo. During run-in and after 7 days of treatment, patients underwent 24 hour Holter monitoring and exercise tests. We analyzed the 24-hour Holter recordings with customized software that computes the correlation between heart rate and occurrence of PVBs. We also computed spectral measures of heart rate variability on the same recordings. Propranolol caused a significant decrease in the log-transformed total number of PVBs recorded over 24 hours and during the day. The number of PVBs was much lower during the night than during the day both after placebo and after propranolol. There were no differences between the two treatments. During the day, there was a positive correlation between heart rate and the number of PVBs in all 18 patients. The mean correlation coefficients between heart rate and number of PVBs increased significantly after propranolol treatment both during the 24-hour monitoring (p < 0.05) and during the day (p < 0.05). The night-recorded correlation coefficients between heart rate and number of PVBs were not significantly different in the placebo versus propranolol group. Propranolol significantly increased the total power during the day. Placebo caused a significant decrease in the low-frequency band (LF) and a significant increase in the high-frequency band (HF) during the night compared with the day. During the day, propranolol significantly reduced LF power and increased HF power, with respect to placebo. After propranolol treatment, the values of LF and HF power during the day were comparable to those recorded at night. The LF/HF ratio decreased significantly after propranolol treatment with respect to placebo in the day and became similar to that recorded during sleep. Propranolol significantly reduced heart rate and systolic blood pressure at rest and at peak exercise and reduced signs of myocardial ischemia. Propranolol administration reduces PVBs in patients with coronary artery disease and severe ventricular arrhythmias possibly through an improvement of cardiac autonomic regulation and through anti-ischemic effects, antiarrhythmic effects, or both. PMID- 11486157 TI - Abnormal gene expression of cardiac fibroblasts in experimental aortic regurgitation. AB - Extensive primary fibrosis precedes heart failure and death in experimental chronic aortic regurgitation. To seek the molecular basis for this observation, this study analyzed the RNA pool for genes that are up-or downregulated in aortic regurgitation fibroblasts. Differential display reverse transcriptase polymerase chain reaction was used to compare RNA extracted from cardiac fibroblasts isolated from three healthy New Zealand white rabbits and from three with aortic regurgitation. Using two base anchoring oligo d(T) primers (T11VN) together with arbitrary upstream primers, numerous differences in normal versus aortic regurgitation gene expression were apparent on differential display reverse transcriptase polymerase chain reaction. The aortic regurgitation cell cultures showed numerous differentially up-and downregulated genes compared with cell cultures of normal cardiac fibroblasts. The results showed that pathologic fibrosis in chronic experimental aortic regurgitation is associated with abnormal cardiac fibroblast gene expression, which may be pathogenic for the fibrous lesion. PMID- 11486158 TI - Efficacy of low-dose amiodarone in the prevention of paroxysmal atrial fibrillation resistant to type IA antiarrhythmic drugs. AB - The efficacy and safety of low-dose amiodarone (Cordarone; Wyeth-Ayerst, Philadelphia, PA) was assessed in 62 symptomatic patients with paroxysmal atrial fibrillation who were resistant to at least two types of IA drugs. The beneficial response to this treatment was defined as a reduction in paroxysmal atrial fibrillation of greater than or equal to 50% within 1 month. Of the 42 patients (67.7%) who were responders, 39 (62.9%) were completely free of episodes. Intolerable side effects were seen in 12 patients (19.3%). Tolerable side effects were encountered by 73% of patients. Most of the adverse effects were transient and responded to a reduction in the dose. In conclusion, (1) low-dose amiodarone produces a beneficial response in the prevention of paroxysmal atrial fibrillation, and (2) low-dose amiodarone is well tolerated. PMID- 11486159 TI - A prospective study of pain control in the emergency department. AB - The most common complaint in the emergency department is pain. The management of acute pain, however, has not been well studied. This prospective study was designed to assess pain intensity and relief along with satisfaction in the emergency department. Adult patients with a primary complaint of acute pain were asked to complete a two-part questionnaire administered by a research assistant. The first part was completed on arrival and the second part on discharge from the emergency department. The respondents were not permitted to see the first part of the questionnaire while completing the second. The questionnaire used an unmarked, horizontal 10-cm visual analog scale along with short answer questions to measure pain, relief, and satisfaction. Choice of drug therapy was decided by the physician according to usual treatment methods. Fifty-seven people presented with the chief complaint of pain. Of those, 30 (53%) were treated with medications. The mean level of pain on admission for treated patients was 6.64 compared with a mean level of pain on discharge of 4.02 (P =.0001). Untreated patients had a mean admission visual analog scale score of 4.19. Compared with treated patients, this difference was statistically significant (P =.001). A mean visual analog scale score of 5.43, representing the mean amount of pain relief, was reported among treated patients. Treated patients also reported a visual analog scale score of 6.46 in overall satisfaction with pain management. The results of this study indicate that there is a significant and clinical difference in levels of pain and satisfaction between admission and discharge in these patients in the emergency department. PMID- 11486160 TI - FDA censoring of manufacturers' postmarketing commitments in HIV/AIDS drug approval letters. AB - CONTEXT: Changing evidentiary standards and partial shift of the investigational phase of drug approval process to the postmarketing phase. OBJECTIVE: To determine the availability of information for independent researchers needed to examine accelerated drug approvals to determine how they differ from traditional drug approvals in the HIV/AIDS domain. DESIGN: Identification of all approved HIV/AIDS and AIDS-related conditions drugs between 1987 and 1999. Follow-up of postmarketing study requirements in the approval letters addressed to the manufacturers. SETTING: Accelerated approval has been expanded to other disease conditions in the past decade. INTERVENTION: Request of approval letters from the U.S. Food and Drug Administration for 76 regulatory actions including expanded access and accelerated and traditional approvals for 42 drugs under the Freedom of Information Act (FOIA) between September 1998 and October 1999. MAIN OUTCOME MEASURE(S): Obtainability of approval letters and uncensored postmarketing study requirements. RESULTS: Fifty-five approval letters were received. Postmarketing study commitments of manufacturers were censored in 25 letters received. We were unable to obtain uncensored copies of those approval letters as of May 2000. Censoring was associated with whether (1) the Prescription Drug User Fee Act of 1992 was applicable to the drug (odds ratio (OR) = 5.7, 95% confidence interval (CI) = 1.4- 23.7) and (2) the new drug application was for a new molecular entity or new drug formulation (OR = 4.2, 95% CI = 1.3 -13.6). CONCLUSIONS: Continued secrecy may stifle independent research and hinder health care providers and patients in making informed decisions. PMID- 11486161 TI - University of Miami Division of Clinical Pharmacology therapeutic rounds: irritable bowel syndrome-pathophysiology, diagnosis, and treatment. AB - Irritable bowel syndrome, or IBS, is a common disorder of the bowels that causes significant patient morbidity, work-absenteeism, physician visits, and, frequently, extensive evaluations. The syndrome has been classified as a functional disorder because of the lack of clearly elucidated pathophysiology, although altered visceral sensitivity, intestinal motility, and psychosocial factors all likely play a role. Treatment includes fostering a strong, trusting relationship with the patient, with pharmacotherapy directed at the predominant symptoms. The authors review the pathophysiology, diagnosis, and treatment of this difficult disorder. PMID- 11486162 TI - Treatment of secondary lymphedema of the arm with physical decongestive therapy and sodium selenite: a review. AB - Secondary lymphedema (LE) in the proximal extremities develop with relatively high frequency in cancer patients after tumor resection, lymph-node obliteration, and/or postoperative irradiation. Physical therapy combined with manual or mechanical lymph drainage and compression bandaging provides symptomatic relief but does prevent the progression of degenerative changes in the affected tissues. As biochemical studies have linked these changes significantly to the excessive generation of oxygen radicals in the affected tissues, LE therapy should aim to eliminate oxygen radical production. Because selenium is a functional component of antioxidant enzymes, has anti-inflammatory properties, and reduces the expression of endothelial cell adhesion molecules, its effect was investigated in postmastectomy patients with LE of the arm. Sodium selenite administered orally in isotonic solution (selenase) at oral dosages of 800 microg Se/day on days 1 through 4 and 500 microg Se/day on days 5 through 28 produced a spontaneous reduction in LE volume and normalized blood parameters in a manner consistent with diminished oxygen radical production. In a randomized, placebo-controlled, double-blind study with postmastectomy LE patients undergoing combined physical decongestion therapy (CPDT), selenite at similar dosages increased the efficacy of CPDT and improved the mobility and heat tolerance of the affected extremity. The patients in this study received 1000 microg of Se/day orally during the first week, 300 microg Se/day during the second and third weeks, and a maintenance dose of 100 microg Se/day during 3 months of follow-up. All patients remained erysipelas-free during the 3 weeks of CPDT and the 3-month follow-up period. Based on the available evidence, supplementation with sodium selenite in isotonic solution is judged to be a valuable and safe extension of the physical decongestive therapy of LE. PMID- 11486163 TI - Pathology manpower: a few rays of sunshine. PMID- 11486164 TI - Trends in pathology graduate medical education. AB - Comprehensive data show trends in graduate medical education in pathology with regard to the numbers of accredited programs, persons certified from those programs, and demographics of the population of first year-trainees in pathology. Experience with US seniors and foreign-trained physicians in the PGY match process for pathology from 1991 through 2000 is presented, along with data on the types of medical schools generating pathology trainees for the PGY-1 year and the top medical schools of origin of US medical graduates who completed the program and became certified in pathology between 1995 and 1999. The impact of reimbursement of the credentialing year is also addressed through data collected from the PRODS Survey 2000, and those results are reviewed. Finally, turnover rates among pathology program directors of combined AP/CP programs and subspecialty programs since 1994 are presented. An analysis of these trends is provided, along with suggestions to improve both the perception of careers in pathology and the actual choice of a career in pathology. PMID- 11486165 TI - A tale of two systems: Pathology resident recruitment in and out of the National Resident Matching Program. AB - The recruitment of most first-year pathology residents currently occurs via 2 systems: the National Resident Matching Program (NRMP), known as the Match, and an underground Out-of-Match process. Out-of-Match recruitment is not a sanctioned option for American medical school seniors. Recent declines in American seniors choosing pathology has intensified pressures to recruit good international medical graduates (IMGs). Pressures by programs on IMGs and by IMGs on programs result in the Out-of-Match recruitment of many IMGs who initially enroll in the NRMP. IMGs may be offered unsolicited Out-of-Match positions with a decision deadline even though they are still interviewing. Some are told if they decline an offer, they will not be ranked by that program in the NRMP process. To enhance apparent recruitment success, programs also feel pressured to participate in the Out-of-Match process, as well as offer positions before they have interviewed all applicants. Coercion of applicants and programs contravenes the spirit of the NRMP, compromises the ability of applicants and programs to best match needs, taints constructive interactions among program directors, and does not enhance pathology's image among applicants or in the broader medical community. PMID- 11486166 TI - Primary neuroendocrine (Merkel cell) carcinoma of the skin: morphologic diversity and implications thereof. AB - A significant proportion of primary neuroendocrine cell carcinomas of the skin (Merkel cell carcinomas [MCCs]) have been reported to occur in intimate association with malignant epithelial neoplasms, mainly squamous cell carcinomas. In addition, divergent differentiation within these tumors, particularly of squamous and eccrine types, is not infrequent. This expanded morphologic spectrum of MCC calls for evaluation of potential biologic implications of the phenotypic diversity and begs reconsideration of the histogenesis of the lesion. The current retrospective review of 29 cases of primary cutaneous neuroendocrine cell carcinoma aims to address these issues by integrating new information with that which is extant. Eleven tumors were associated with evolving or established cutaneous carcinomas: 2 actinic keratoses, 5 Bowen's disease, 3 superficial squamous cell carcinomas, and 1 basal cell carcinoma. Two combined squamous neuroendocrine tumors occurred in recipients of solid organ transplants, and another developed in a Marjolin's ulcer at the site of a previous burn. Squamous and/or adnexal differentiation within the dermal component of the tumor was observed in 4 instances and was significantly associated with MCCs in intimate association with another cutaneous carcinoma. The outcome of the group as a whole is similar to that recorded in previous series of MCC, with local recurrence in 32% of cases and death caused by the neoplasm in 28%. Only 52% of the patients were alive with no history of metastasis at follow-up. No significant difference in outcome was observed between the patients with pure MCCs and those with MCCs in combination with another cutaneous carcinoma. PMID- 11486167 TI - E-cadherin expression in prostate cancer: a broad survey using high-density tissue microarray technology. AB - E-cadherin is a calcium 2+-dependent cell-adhesion molecule that determines epithelial development in the embryo and maintains adult differentiated epithelium and homeostasis. Aberrant or decreased expression has been reported to be associated with prostate carcinoma progression. The degree of E-cadherin expression in prostate cancer remains controversial. Some studies have reported decreased expression of E-cadherin as tumors advance and metastasize. Other studies have not demonstrated this relationship. To address these variations, we undertook a study to systematically evaluate E-cadherin expression in a broad range of prostate tissue. Benign prostate, clinically localized prostate cancer, and hormone-refractory metastatic prostate cancer were analyzed under uniform conditions using high-density tissue microarrays (TMA). Formalin-fixed, paraffin embedded prostate carcinoma from men with clinically localized prostate carcinoma and autopsy material from men who died of widely metastatic, hormone-refractory prostate carcinoma were arrayed into 6 high-density TMA blocks. Benign and atrophic prostate tissue and high-grade prostatic intraepithelial neoplasia (PIN) were also included from the clinically localized cases. Immunohistochemistry was performed using the immunoglobulin G1 mouse monoclonal antibody (HECD-1; Zymed, San Francisco, CA). Membranous staining was recorded as low (aberrant) or high (normal). E-cadherin expression was considered aberrant if less than 70% of the cells had strong membranous staining. A total of 1,220 prostate TMA samples were analyzed. High (normal) E-cadherin expression was seen in 87% of 757 benign, 80% of 41 high-grade PIN, 82% of 325 prostate carcinoma and 90% of 97 hormone refractory prostate carcinoma TMA samples. Mean E-cadherin expression was determined for each of the 128 clinically localized prostate cancer cases. Aberrant E-cadherin expression showed a statistical trend toward an association with positive surgical margins (P =.012), higher Gleason score (P =.18), and prostate-specific antigen (PSA) failure (Kaplan-Meier analysis, log-rank P =.09). There was a statistically significant association between aberrant E-cadherin expression and larger tumor size (P =.01). No significant associations were seen with extraprostatic extension and seminal vesicle invasion. The current study shows a broad-spectrum approach to evaluating E-cadherin protein expression in prostate carcinoma. Clinically localized prostate tumors, treated with surgery alone, show a high level of E-cadherin expression. Aberrant expression was identified in tumors with positive surgical margins, higher Gleason score, and a higher rate of PSA failure. However, these trends were not statistically significant. A statically significant association between aberrant E-cadherin expression and larger tumor size was identified. In the metastatic hormone refractory prostate tumors, E-cadherin expression was vastly expressed, and only rare cases had aberrant expression. Therefore, the findings of this study are most consistent with a transient down-regulation of E-cadherin in localized prostate cancer. Metastatic prostate cancer shows strong E-cadherin expression as determined by anti-E-cadherin antibody HECD-1. PMID- 11486168 TI - Genomic instability in chronic viral hepatitis and hepatocellular carcinoma. AB - Chronic hepatitis may progress to cirrhosis and hepatocellular carcinoma (HCC). Progressive accumulation of mutations and genomic instability in chronic viral hepatitis might flag an increased risk of HCC development. Genomic instability at dinucleotide microsatellite loci in chromosomes 2, 13, and 17 and at 2 mononucleotide repeat loci was examined in liver tissues from 41 patients, including 30 without HCC (18 patients with chronic hepatitis and 12 with cirrhosis) and 11 with HCC. Genomic instability was detected in 51% of the 41 cases. Allelic imbalance at informative dinucleotide loci occurred in 37% of the cases. In 14 cases (34%), allelic imbalance was detected in chronic hepatitis or cirrhosis without HCC. Allelic imbalance at the chromosome 13 locus was detected in 50% of the cases of chronic hepatitis C. Allelic imbalance at the TP53 chromosome locus and/or at the chromosome 13 locus was significantly more frequent than alterations at the chromosome 2 locus (P =.026). Low-level microsatellite instability was found in 20% of all cases examined and high-level microsatellite instability in 3 patients (7.5%), including 2 cases of chronic hepatitis and 1 case of cirrhosis. Our results show that allelic imbalance occurs frequently in hepatitis-related HCC as well as in chronic hepatitis in patients without HCC. Allelic imbalance at the D13S170 chromosome 13 locus (13q31.2) occurs frequently in chronic hepatitis, suggesting that genomic alterations affecting the long arm of chromosome 13 might be used to monitor the natural progression of chronic hepatitis-associated liver carcinogenesis. PMID- 11486169 TI - Superficial acral fibromyxoma: a clinicopathologic and immunohistochemical analysis of 37 cases of a distinctive soft tissue tumor with a predilection for the fingers and toes. AB - This report describes the clinicopathologic features and immunohistochemical findings identified in 37 cases of a distinctive soft tissue tumor that has a predilection for the hands and feet. The study group included 25 male and 12 female subjects ranging in age from 14 to 72 (mean, 43; median, 46) years. The patients presented with solitary masses 0.6 to 5.0 cm (mean, 1.75 cm) that were present from 3 months to 30 years (median duration, approximately 3 years) before surgical intervention and involved the toes (n = 20), fingers (n = 13), and palm (n = 4). Twenty of the cases were documented to involve the nail region. Histologically, the tumors were typically located in the dermis or subcutis and composed of spindled and stellate-shaped cells with random, loose storiform, and fascicular growth patterns. The lesional cells were embedded in myxoid or collagenous matrix, often with mildly to moderately accentuated vasculature and increased numbers of mast cells. There was generally slight to mild nuclear atypia; only 3 cases had more substantial atypia. Mitotic figures were infrequent. Occasional multinucleated stromal cells were noted in 19 cases. The process showed immunoreactivity for CD34 (21 of 23 cases), epithelial membrane antigen (18 of 25 cases), and CD99 (11 of 13 cases). No immunoreactivity was detected for actins, desmin, keratins, or HMB-45, and only 1 of 23 tumors had weak reactivity for S100 protein. The surgical specimens consisted of biopsy or partial resection specimens (n = 4), local excisions (n = 29), and amputated or partially amputated digits (n = 4). Detailed follow-up, available for 18 patients (mean follow-up interval, 10.1 years), revealed 1 recurrence after local excision and 2 instances of persistent or progressive disease after partial excision. A differential diagnosis of fibrous histiocytoma, dermatofibrosarcoma protuberans, acquired (digital) fibrokeratoma, sclerosing perineurioma, cutaneous myxoma (superficial angiomyxoma), and acral myxoinflammatory fibroblastic sarcoma is discussed. PMID- 11486170 TI - Superficial cervicovaginal myofibroblastoma: fourteen cases of a distinctive mesenchymal tumor arising from the specialized subepithelial stroma of the lower female genital tract. AB - The clinicopathologic features and immunohistochemical profiles of 14 cases of a distinctive mesenchymal tumor that arises in the superficial lamina propria of the cervix and vagina and is histologically distinguishable from mesodermal (fibroepithelial) stromal polyp, including the cellular (pseudosarcomatous) variant, angiomyofibroblastoma, aggressive angiomyxoma, and other well-recognized lesions that occur in this location, are described. The lesions presented as a polypoid (n = 10) or nodular (n = 4) mass in the vagina (n = 12) or cervix (n = 2) of women ranging in age from 40 to 74 years (median, 58 years). The tumors were subepithelial in location, were well circumscribed, and ranged in size from 1 to 6.5 cm. (mean, 2.7 cm). Microscopically, the process was moderately to highly cellular and composed of relatively bland spindled and stellate-shaped mesenchymal cells embedded in a finely collagenous stroma that was punctuated by myxoid and edematous foci in 9 cases. The lesions characteristically had a multipatterned architecture with tumor cells focally assuming a lacelike/sievelike growth pattern in the more stroma-rich areas of the tumor and a vague fascicular growth pattern in the more cellular foci. Mitotic activity was minimal, and no atypical mitotic figures were identified. The tumors were immunoreactive (in decreasing order of relative strength) for vimentin (5 of 5 cases), estrogen (10 of 10 cases), and progesterone (10 of 10 cases) receptors, desmin (13 of 13 cases), CD34 (11 of 13 cases), alpha-smooth muscle actin (5 of 11 cases), and muscle-specific actin (2 of 8 cases). The desmin and CD34 antibodies highlighted the interconnecting, dendritic processes associated with many of the tumor cells. No immunoreactivity was detected for S100 protein, epithelial membrane antigen, or keratins. Follow-up data for 11 patients (range, 1 to 20 years; median, 4 years) showed no recurrence or metastasis after local excision. The term "superficial cervicovaginal myofibroblastoma" is proposed because it reflects the distinguishing features of this benign, relatively site specific mesenchymal tumor. The process probably arises as a neoplastic proliferation of hormonally responsive mesenchymal cells native to the unique subepithelial stromal layer normally found through the endocervix and vulva of adult women. PMID- 11486171 TI - Rosette-forming epithelioid osteosarcoma: a histologic subtype with highly aggressive clinical behavior. AB - Osteosarcoma shows a variety of histologic patterns. Rarely, this tumor may appear epithelioid, including a rosettelike configuration simulating glands. We retrospectively reviewed 16 cases of osteosarcoma with rosettelike structures treated at the National Cancer Center and Akita University, Japan, from 1972 to 1999. The 16 patients were under 30 years of age, and males were predominant in the sex distribution. The tumors arose primarily in the metaphysis of long tubular bones, and the most common symptom was pain. Roentgenographically, the lesions showed a highly destructive appearance with varying degrees of mineralization. Twelve patients (75%) died of multiple lung metastases in spite of surgery with wide surgical margins and systemic chemotherapy. The estimated cumulative 5-year survival rate was 15%, significantly worse than the rate of 55% in 70 cases of conventional osteoblastic osteosarcoma without rosettelike structures arising in long tubular bones. All of the 16 tumors, originally classified as conventional osteoblastic osteosarcoma, predominantly displayed a small multinodular growth pattern with lacelike osteoid deposits in the center between dilated blood vessels showing a hemangiopericytoma-like appearance. Ten tumors (63%) showed immunoreactivity for epithelial membrane antigen. We believe rosette formation in osteosarcomas of long tubular bones is an ominous sign; therefore, those tumors should be distinguished from osteosarcomas with conventional morphotypes. PMID- 11486172 TI - Adefovir nephrotoxicity: possible role of mitochondrial DNA depletion. AB - This report investigates the pathomechanism of acute renal failure caused by toxic acute tubular necrosis after treatment with the antiretroviral agent adefovir. A 38-year-old white homosexual man with human immunodeficiency virus infection and no history of opportunistic infections was maintained on highly active antiretroviral therapy (HAART), including hydroxyurea, stavudine, indinavir, ritonavir, and adefovir dipivoxil. Histologic examination of the renal biopsy showed severe acute tubular degenerative changes primarily affecting the proximal tubules. On ultrastructural examination, proximal tubular mitochondria were extremely enlarged and dysmorphic with loss and disorientation of their cristae. Functional histochemical stains for mitochondrial enzymes revealed focal tubular deficiency of cytochrome C oxidase (COX), a respiratory chain enzyme partially encoded by mitochondrial DNA (mtDNA), with preservation of succinate dehydrogenase, a respiratory chain enzyme entirely encoded by nuclear DNA (nDNA). Immunoreactivity for COX subunit I (encoded by mtDNA) was weak to undetectable in most tubular epithelial cells, although immunoreactivities for COX subunit IV and iron sulfur subunit of respiratory complex III (both encoded by nDNA) were well preserved in all renal tubular cells. Single-renal tubule polymerase chain reaction revealed marked reduction of mtDNA in COX-immunodeficient renal tubules. We conclude that adefovir-induced nephrotoxicity is mediated by depletion of mtDNA from proximal tubular cells through inhibition of mtDNA replication. This novel form of nephrotoxicity may serve as a prototype for other forms of renal toxicity caused by reverse transcriptase inhibitors. PMID- 11486173 TI - Angiocentric T-cell lymphoma of the pancreas presenting as late-onset diabetes mellitus with diabetic retinopathy. AB - Pancreatic lymphoma presenting with clinical diabetes mellitus (DM) is rare. We report angiocentric T-cell lymphoma of the pancreas in a 65-year-old Thai woman who presented with progressive deterioration of visual acuity of both eyes. She had diabetic retinopathy (DR) diagnosed only 20 months after the diagnosis of DM at the age of 63. She later developed erythematous rashes, fever, and deterioration of consciousness; she eventually died of shock. A skin biopsy and bone marrow examination revealed angiocentric T-cell lymphoma. At autopsy, the pancreas and both eyes were extensively infiltrated by lymphoma. Widespread involvement of nearly all organs but superficial lymphadenopathy was detected. In contrast to other typical cases of long-standing DM, only mild atherosclerosis was noted, and no DR was found. To the best of our knowledge, this is the first case of lymphoma involving the pancreas and both eyes producing clinical DM and DR. PMID- 11486175 TI - Leptospirosis mimicking acute cholecystitis among athletes participating in a triathlon. AB - Leptospirosis, a disease acquired by exposure to contaminated water, is characterized by fever accompanied by various symptoms, including abdominal pain. An acute febrile illness occurred in athletes who participated in an Illinois triathlon in which the swimming event took place in a freshwater lake. Of 876 athletes, 120 sought medical care and 22 were hospitalized. Two of the athletes had their gallbladders removed because of abdominal pain and clinical suspicion of acute cholecystitis. We applied an immunohistochemical test for leptospirosis to these gallbladders and demonstrated bacterial antigens staining (granular and filamentous patterns) around blood vessels of the serosa and muscle layer. Rare intact bacteria were seen in 1 case. These results show that leptospirosis can mimic the clinical symptoms of acute cholecystitis. If a cholecystectomy is performed in febrile patients with suspicious environmental or animal exposure, pathologic studies for leptospirosis on formalin-fixed, paraffin-embedded tissues may be of great value. PMID- 11486174 TI - Follicular dendritic cell tumor presenting in the lung: a case report. AB - An example of extranodal follicular dendritic cell sarcoma (FDCS) presenting in the lung, a heretofore unreported site, is described. Macroscopically, a 9.5-cm, tan-white, dominant mass and multiple smaller parenchymal and pleural nodules were identified. Microscopically, the tumor was composed of spindled cells with uniform cytologic features arranged in short, intersecting fascicles and intermixed small lymphocytes and plasma cells. One of 4 peribronchial and hilar lymph nodes evaluated microscopically was focally involved by the process. Immunohistochemically, the neoplastic spindled cells expressed complement receptors CD21 and CD35 and low-affinity nerve growth factor receptor but did not express keratin (AE1/AE3 and CAM5.2), CD45 (leukocyte common antigen), CD20 (L26), S-100 protein, muscle-specific actin, or gp100 protein (HMB45). Ultrastructurally, the tumor cells have complex interdigitating cell surface processes and desmosomes. Epstein-Barr virus (EBV) was not detected in the tumor cells by in situ hybridization for EBV-encoded RNA or by polymerase chain reaction for viral DNA. FDCS should be considered in the differential diagnosis of any spindled-cell tumor with interspersed chronic inflammatory cells occurring in the lung. An immunohistochemical panel, including anti-CD21 and -CD35, can assist in its diagnosis, especially with small bronchial biopsy specimens. 2001 by W.B. Saunders Company. PMID- 11486176 TI - Primary mammary small-cell carcinoma: a molecular analysis of 2 cases. AB - Primary small-cell carcinoma of the breast is an exceedingly rare variant of breast carcinoma whose genetic profile has not been previously investigated. We report the molecular features of 2 cases of small-cell carcinoma of the breast: 1 with an adjacent intraductal carcinoma, and 1 with prior pleomorphic lobular carcinoma in situ. Laser capture microdissection followed by loss of heterozygosity (LOH) analysis revealed identical molecular alterations at multiple chromosomal regions, including BRCA-1, BRCA-2, p53, and retinoblastoma gene loci, in 1 case of small-cell carcinoma and its adjacent intraductal component. Additionally, LOH in 1 or both small-cell carcinomas was detected at 3p, 4q31.2-qter, 8p21-24, 11q13 (MEN-1 locus), 11q23.3, 11q24.1-25, 16q24.1 (H cadherin locus), and 17q25. The results of our molecular analysis suggest that genetic changes in mammary small-cell carcinoma resembled those seen in both invasive ductal carcinomas and pulmonary small-cell carcinoma. Second, mammary small-cell carcinoma is clonally related to ductal carcinoma in situ and might represent an example of divergent differentiation occurring in a multipotential neoplastic stem cell. PMID- 11486177 TI - Severe hepatitis caused by Epstein-Barr virus without infection of hepatocytes. AB - Although hepatitis is a common feature of primary Epstein-Barr virus (EBV) infection, severe liver injury is rare and its pathogenesis is unclear. A previously healthy girl developed severe hepatitis with prolonged jaundice. Serologic examination showed that she had primary infection with EBV. An extremely high Epstein-Barr viral load was observed in her peripheral blood. The viral load decreased in parallel with symptomatic improvement. Histologic examinations showed spotty necrosis of the liver parenchyma and infiltration by CD8(+) T cells. The CD8(+) T cells, not hepatocytes, were positive for EBV. Possible mechanisms of viral hepatitis without infection of hepatocytes are discussed. PMID- 11486180 TI - Preface. PMID- 11486178 TI - The origin of striated muscle cells in non-neoplastic lung tissue. PMID- 11486181 TI - Introduction. PMID- 11486182 TI - Double-outlet right ventricle: Pathology and angiocardiography. AB - Double-outlet right ventricle is but one form of abnormal ventriculoarterial connection. The definition that more than half of each great artery originates above the morphologically right ventricle is arbitrary. As pointed out by Lecompte, those features that should be defined in hearts with the ventriculoarterial connection of double-outlet right ventricle (and indeed other forms of abnormal ventriculoarterial connection) include the nature of the infundibular septum, ventriculoinfundibular fold, trabeculoseptomargin-alis, attachments of infundibular septum to anterior or posterior limb of trabeculoseptomargin-alis, the size and position of the ventricular septal defect, the spatial relation of great artery(s) to the ventricular septal defect, the spatial relationship between the great, and the distance between the tricuspid and pulmonary valves and the semilunar valves. Copyright 2000 by W.B. Saunders Company PMID- 11486183 TI - Double-outlet right ventricle: An echocardiographic approach. AB - Using echocardiography it is possible to obtain detailed information about the basic morphology, subtypes, and associated abnormalities in double outlet right ventricle. Armed with this information, surgical management can be planned in the majority of cases utilizing echocardiography alone. This article addresses the basic echocardiographic approach, provides an in-depth understanding of the various subtypes, as well as outlining the additional features that often complicate complete repair. Copyright 2000 by W.B. Saunders Company PMID- 11486184 TI - Surgical management of double-outlet right ventricle with subaortic ventricular septal defect. AB - Double-outlet right ventricle (DORV) and subaortic ventricular septal defect (VSD) is defined anatomically as a defect where the entire pulmonary trunk and at least half of the aorta arises from the right ventricle (RV) and the VSD is most closely aligned with the aorta. The surgical management of DORV and subaortic VSD usually results in a 2 ventricle repair where the left ventricular outflow is diverted via the VSD to the aorta. Pulmonary and/or subpulmonary obstruction is found in a large portion of these patients and requires relief at the time of repair. Resection of subpulmonary muscular obstruction with right ventricular outflow augmentation with a patch is required in most to eliminate the obstruction. Important anatomic features to consider in constructing the left ventricular outflow tunnel include the prominence of the conal septum, tricuspid valve attachments to conal septum, and the distance between the pulmonary and tricuspid valves. Operative mortality is low and long-term complications and need for reoperation is uncommon. This report describes the surgically relevant anatomic features and operative techniques. Copyright 2000 by W.B. Saunders Company PMID- 11486185 TI - Biventricular repair for double-outlet right ventricle. AB - Double-outlet right ventricle (DORV) is a heart malformation that describes an anomalous ventriculoarterial connection which can be associated with considerable variant of associated lesions. When this malformation is present with two ventricles, biventricular repair is feasible in the vast majority of cases. This report describes the surgical techniques for biventricular repair in all forms of encountered DORV, as well as the surgical strategy employed at our institution. Copyright 2000 by W.B. Saunders Company PMID- 11486186 TI - The role of the Fontan procedure in the surgical treatment of congenital heart malformations with double-outlet right ventricle. AB - The Fontan procedure in one of its many modifications is applicable to the surgical treatment of those patients with double-outlet right ventricle who are not amenable to bi-ventricular correction. Included in this group are patients with significant hypoplasia of one of the ventricles, patients with distant (non committed) ventricular septal defect, patients with significant straddling of the atrioventricular valves, patients with abnormal atrioventricular connections (such as absence of one of the valves), or common atrioventricular valve connection. The presence of complex associated lesions such as those seen in the heterotaxia syndromes may, by virtue of the surgical risk of attempts at bi ventricular repair, make the Fontan approach a suitable option for these patients. The presence of a subaortic conus and the potential for subaortic obstruction, a frequent findings in patients with double outlet right ventricle, plays an important role in the preparation and conduct of the Fontan operation. Copyright 2000 by W.B. Saunders Company PMID- 11486187 TI - Pathobiology of pulmonary hypertension: Impact on clinical management. AB - Our previous studies showed how analysis of pulmonary vascular changes on lung biopsy tissue and on angiography added to the hemodynamic assessment of pulmonary vascular resistance in predicting the success of a surgical repair. Both the potential for heightened vasoreactivity in the early postoperative period and for reversibility of pulmonary vascular disease at later follow-up were correlated with qualitative and quantitative evaluation of arterial changes. The ability of continuous intravenous prostacylin to arrest progression and even induce regression of structurally advanced pulmonary vascular disease in some cases has led to rethinking how pathological material can be useful in clinical decision making. The presence of occlusive changes and particularly plexiform lesions was thought to represent irreversible disease, but the observation that ongoing cellular proliferation and connective tissue synthesis occurs even in advanced lesions thought to represent end stage 'burnt-out' lesions, led to re-evaluation of the potential of biologically reversing the disease process. Our laboratory has used clinical material, cultured cells, and studies in experimental animals to gain new insights into some of the mechanisms which lead to the progression of vascular changes, and has used this information in strategies aimed at arresting progression and, more recently, inducing regression of pulmonary hypertension and associated vascular lesions. Specifically, we have focused on the increased activity of an endogenous vascular elastase (EVE) and expression of the glycoproteins tenascin and fibronectin in the pathobiology of pulmonary hypertension. This report will first review our studies in children with congenital heart defects, assessment of reversibility of pulmonary hypertension, and then discuss more recent work addressing cellular and molecular mechanisms aimed at developing newer therapeutic strategies. Copyright 2000 by W.B. Saunders Company PMID- 11486188 TI - Cardiopulmonary bypass in infants and children. AB - Cardiopulmonary bypass (CPB) systems have evolved from futuristic visions of surgical pioneers to a safe and efficient tool in the therapy of treatment of cardiac disorders. There are many significant differences in the physiology between neonates and adult patients. There are currently very few congenital cardiac malformations that cannot be addressed effectively with surgical therapy. Yet, the necessity of CPB in the repair of these patients can still result in significant morbidity. A clearer understanding of the effects of CPB, hypothermia, and circulatory arrest is evolving and there is a considerable amount of research in these areas. It seems likely that modification of current CPB systems, minimization of exposure, and surgical techniques to avoid or limit the adverse effects may reduce mortality and morbidity in the future. The problems faced in these complex patients and procedures require that infant and neonatal cardiac surgery be performed in specialized centers with a multidisciplinary approach and specialized personnel. Future improvements in technology will likely result in improved long term outcome for children with congenital cardiac disease. Copyright 2000 by W.B. Saunders Company PMID- 11486189 TI - Databases for congenital heart disease: A beginner's guide. AB - We discuss the reasons for collecting data on operations for congenital heart defects. Computerized databases facilitate the collection, storage, and retrieval of such data. We outline the basic criteria for selecting a suitable database to cater to the particular needs of a surgeon or a department and stress the importance of the accuracy of the data held in a database. We explain the relative advantage of simple versus complex database systems. We conclude with a summary of the steps required for a practicing congenital heart surgeon to set up and operate a simple database. Copyright 2000 by W.B. Saunders Company PMID- 11486190 TI - Modified Rastelli using an autograft: A new concept for correction of transposition of the great arteries with ventricular septal defect and left ventricular outflow tract obstruction (with an extension to other congenital heart defects). AB - We describe in this article a new approach in transposition of the great arteries (TGA), ventricular septal defect (VSD), and left ventricular outflow tract obstruction (LVOTO). It consists of an operation combining the intracardiac infundibular resection of the Lecompte procedure to allow a direct VSD-aorta connection and the right ventricle-pulmonary artery (RV-PA) connection with a segment of tubular aortic autograft, (with or without an additional monocusp patch). Thus, a viable conduit, capable of growth, is interposed between RV and PA. We have performed this procedure in 19 patients (age, 2 months to 11 years), 6 being under 1 year. All patients are doing well after a maximum follow-up of 72 months, exhibiting growth of the conduit and satisfactory RV pressures. This new approach allows primary correction even in the young and can be extended, we think, for other defects with pulmonary stenosis or atresia. It shares, however, with many other procedures, the valveless nature of the repair with the potential late need for valve replacement surgery. Copyright 2000 by W.B. Saunders Company PMID- 11486191 TI - Coarctation of the aorta. AB - Controversy still exists in the literature regarding definitive therapy for repair of coarctation of the aorta. Major factors involve not only the timing of repair, but also the method of repair, whether surgical or by percutaneous transluminal balloon dilatation. Results and complications of coarctation repair using various methods of classification present a diversity of results. This report will focus on these issues and attempt to dispel the statement that either one method or the other is the "choice method" of repair for any and all types of coarctation. Also presented is a proposed classification we believe will assist in clarifying the choice of therapy and perhaps improve not only the reporting of results, but also the results themselves. Methods of repair are discussed to provide the surgeon with a complete armamentarium of operations that the surgeon would tailor to the individual anatomicopathological patterns of the patient who presents at the time of surgery. Copyright 2000 by W.B. Saunders Company PMID- 11486192 TI - Total anomalous pulmonary venous drainage associated with complex cardiac anatomy. AB - Patients with total anomalous pulmonary venous drainage (TAPVD) presenting with associated complex cardiac anatomy have a markedly less favorable prognosis when compared to patients with isolated TAPVD. The impact of associated cardiac lesions on the pathophysiology of TAPVD and the wide array of operative strategies for surgical palliation is discussed. Copyright 2000 by W.B. Saunders Company PMID- 11486193 TI - Aortic allografts for left ventricular outflow tract replacement in children. AB - Aortic allografts provide many advantages in children requiring left ventricular outflow tract (LVOT) reconstruction. The low risk of thromboembolic events and freedom from the requirement for anticoagulation are primary benefits. Additionally, excellent hemodynamic results are possible even in the presence of multilevel obstruction. The pulmonary autograft has become the favored approach in most pediatric centers, as the limited longevity of the aortic allograft has now become apparent. However, some children are not candidates for the pulmonary autograft. Thus, the aortic allograft remains a useful aortic valve substitute in children. Using standard aortic root replacement (ARR) or extended aortic root replacement (EARR) techniques, aortic allografts can be used in any circumstance. Young age and small size are predictive of shortened valve longevity and higher operative mortality compared with older children. Reoperation to replace a degenerated aortic allograft can be accomplished safely. Copyright 2000 by W.B. Saunders Company PMID- 11486194 TI - Anomalous origin of the left coronary artery from the pulmonary artery: Successful surgical strategy without assist devices. AB - The objective of this review was to analyze a surgical strategy for children with anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) where aortic implantation was successful without the use of extracorporeal membrane oxygenation (ECMO) or left ventricular assist device (LVAD). From 1989 through 1999, 16 consecutive children underwent aortic implantation for ALCAPA. Median age was 0.45 years. Surgical strategy included bicaval venous cannulation, LV vent, moderate systemic hypothermia (28 degrees C), antegrade cardioplegia with pulmonary artery (PA) occlusion, PA transection, aortic implantation with a large "button" of PA, PA reconstruction with pericardium, intraoperative transesophageal echocardiography, and inotropic support with dopamine, dobutamine, and milrinone. All patients survived. No patient was placed on ECMO or LVAD. Mean cardiopulmonary bypass (CPB) time was 152 +/- 41 minutes. Mean aortic cross-clamp time was 46 +/- 13 minutes. Mean time from cross-clamp removal to CPB off ("weaning" time) was 58 +/- 20 minutes. One patient had delayed sternal closure. Only 2 patients required more than 10 &mgr;g/kg/minute of dopamine and dobutamine. Mean hospital stay was 29 +/- 25 days. One patient required outpatient inotropic support for 5 months. Although ECMO or LVAD may be necessary for some ALCAPA patients, we describe a surgical strategy of aortic implantation that was successful without the use of assist devices. Copyright 2000 by W.B. Saunders Company PMID- 11486195 TI - One and one half ventricle repairs. AB - Patients with functionally or anatomically borderline pulmonary ventricles can be managed by a biventricular repair, a Fontan procedure, or by an intracardiac repair in association with a cavopulmonary anastomosis. The latter repair is known as a one and one half ventricle repair. Extending the limits of a biventricular repair can be associated with a high early mortality. The Fontan is associated with late failure. One and one half ventricle repairs are an attempt to reduce early risk and late failure while still achieving separate pulmonary and systemic circulations. The two most common reasons for a one and one half ventricle repair are a small pulmonary ventricle and a dilated poorly functioning pulmonary ventricle. Estimated ventricular volumes of 30% to 80% and z values as small as -10 have been successfully repaired with the one and one half ventricle approach. Ebstein's anomaly is the most common situation where dilated and poorly functioning pulmonary ventricles are treated with a one and one half ventricle repair. There are also special situations where the one and one half ventricle repair is logical. Mortality has ranged from 0% to 12%. No intermediate term sequelae of protein loosing enteropathy, atrial arrhythmias, or pulmonary arteriovenous fistula have been identified. Successful one and one half ventricle repairs have primarily been performed for small pulmonary ventricles and poorly functioning pulmonary ventricles such as those seen with Ebstein's anomaly. Intermediate term follow up has been favorable when compared to the Fontan circulation. Copyright 2000 by W.B. Saunders Company PMID- 11486196 TI - Mechanical support of the pediatric cardiac patient. AB - Increased understanding of the anatomical nuances of congenital heart defects, improved methods of myocardial preservation, and advances in surgical techniques have led to improved results and need for postoperative mechanical support in patients undergoing congenital heart surgery. However, there remains a small portion of patients with myocardial or pulmonary failure that can can be rescued by intelligent use of mechanical support. The most widely used form of mechanical support is extracorporeal membrane oxygenation (ECMO), which has been adapted for use in congenital heart patients as well as patients with respiratory failure. Recent reports have suggested that ventricular support devices may also be useful in this patient population. A promising new application of mechanical support is for rescue during cardiac arrest; surprisingly good results have been obtained in this obviously moribund group of patients. Future developments in mechanical support include more accurate identification of patients who will benefit from this therapy and technological advances that will provide greater biocompatibility and simplification of the support circuit. Copyright 2000 by W.B. Saunders Company PMID- 11486197 TI - Intermediate outcome after Mustard and Senning procedures: A study by the Congenital Heart Surgeons Society. AB - Although atrial switch is rarely performed today except as part of a "double switch" operation, there continues to be interest in the long-term outcome of the procedure because of the many Mustard and Senning survivors who are in follow-up. In contrast to most other reported series on atrial switch, this study by the Congenital Heart Surgeons Society (CHSS) is a prospective multiinstitutional study of patients encountered in a relatively short time frame during the late 1980s. Between 1985 to 1989, 20 centers with surgeons belonging to the CHSS prospectively registered patients <15 days of age presenting with simple or complex transposition of the great arteries (TGA). The patients were assigned to protocol groups based on intent to treat (arterial switch, Mustard, or Senning). Data were abstracted in the Data and Analysis Center, which in most cases, conducted the annual follow-up. Among the 281 patients who had an atrial switch, there were 108 Mustard and 173 Senning procedures. For the combined atrial switch population, the survival at 1 month, 5 years and 10 years was 90%, 85%, and 84%, respectively. Results for the Mustard were better than for the Senning with survival at 1 month, 5 years, and 10 years being 96% versus 86%, 95% versus 80%, 93% versus 78%; (P <.001) for Mustard versus Senning. While the major mortality risk occurred in the first postoperative month for both groups, thereafter, the late rate of death from 1 to 10 years after operation was 0.78%/year Senning versus 0.23%/year Mustard (P <.05). TGA/ventricular septal defect (VSD), lower weight, younger age, cardiac positional anomalies, and procedures on the left ventricle (LV) outflow all correlated significantly with poor outcome. There were 19 reoperations including 2 for right ventricle (RV) failure, 12 for pathway obstruction, and 5 for baffle leak with a 36% overall mortality. Freedom from any pathway obstruction at 10 years was 95%. A permanent pacemaker was required in 21 patients with the significant risk factors including TGA/VSD and a Senning, and previous atrial septectomy. Freedom from a pacemaker insertion at 10 years was 91%. For both Mustard and Senning, 60% of patients were in New York Heart Association (NYHA) functional Class I with the remaining in Class II. This study of atrial switch outcomes by the CHSS suggests that the subgroup undergoing a Mustard procedure had better early and late survival than those undergoing a Senning operation. The overall incidence of baffle pathway obstruction was low though reoperation for this complication carried high risk. Serious arrhythmia requiring medication and the need for a permanent pacemaker were both relatively uncommon. The incidence of severe right heart failure was very low, although only 60% of patients are unrestricted in their activity. Copyright 2000 by W.B. Saunders Company PMID- 11486198 TI - Left ventricular reconditioning and anatomical correction for systemic right ventricular dysfunction. AB - The morphologically right ventricle (mRV) fails after managing systemic workload for a variable period of time in some patients with transposition of the great arteries (TGA) who have undergone an atrial switch operation and in patients with congenitally corrected transposition of the great arteries (cc-TGA). Conventional therapy for progressive mRV failure, including tricuspid valve replacement and cardiac transplantation, has been disappointing. Anatomical correction, reincorporating the morphologically left ventricle (mLV) into systemic circulation, was performed in a total of 84 patients (39 TGA, 45 cc-TGA) in 2 institutions (Royal Children Hospital 1981-1993; Cleveland Clinic Foundation 1993 1999). The mVL was retrained to generate systemic pressure by means of pulmonary artery band in 43 patients. The overall mortality for patients entering this program is 15.4% (8 early and 5 late operative deaths). All of the operative deaths were patients with TGA and a prior atrial switch operation. Four patients (5%) have been transplanted or are on a transplantation waiting list. In patients with TGA and previous atrial switch operation, older age and abnormal coronary anatomy was associated with a higher operative mortality. None of the measured parameters of the mLV and mRV were predictors of failure of mLV reconditioning. Follow-up echocardiographic evaluations of the 46 survivors of anatomical correction showed normal mRV function in 41 (89%) and normal mLV function in 42 patients (91%). mLV retraining and anatomical correction produces good results in prepubescent patients. The response of older patients is less predictable and associated with a higher early and late mortality. Copyright 2000 by W.B. Saunders Company PMID- 11486199 TI - Tetralogy of Fallot: Late outcome after repair and surgical implications. AB - Cardiac surgery has transformed the outcome for patients with tetralogy of Fallot. Repair has conveyed excellent long-term results with most patients remaining well and leading normal lives. However, there are problems with late morbidity and mortality primarily due to right ventricular dysfunction, exercise intolerance, arrythmia, and sudden cardiac death. There has been a dynamic shift in our surgical approach to managing patients with tetralogy over the past 5 decades. This in part accounts for persisting difficulties in predicting late outcome for evry single patient with repaired tetralogy of Fallot. There are, however, several confounding variables, influencing long-term outcome for these patients, namely the underlying anatomical substrate, age at repair, surgical approach to repair, and residual hemodynamic abnormalities. It is gratifying to see that recent knowledge accumulated from long-term follow-up studies is influencing contemporary surgical practice. Individualized strategies aiming to minimize the potential for free pulmonary regurgitation, and the long-term detrimental effects associated with it, need to continue to develop. Preservation of right ventricular and pulmonary valve function combined with early restoration of normal pulmonary blood flow are likely to convey an even better long-term outlook for these patients. Further follow-up studies with assessment of bi venticular function, however, are needed in both our older and contemporary cohorts with repaired tetralogy of Fallot. Copyright 2000 by W.B. Saunders Company PMID- 11486200 TI - The emerging adult population with congenital heart disease. AB - The successes in managing infants and children with congenital heart disease have led to an emerging population of adult patients. As we enter this new century, the majority of patients with congenital heart disease will be adults, not children. It is important to maintain our commitment for continuing care to the emerging adult population. Psycho-social issues, including employment and pregnancy counseling, are required as well as the ongoing need for medical and occasionally surgical intervention. The health care system needs to develop supra regional tertiary referral centers for care of these patients and provide information sharing and support for community-based physicians interested in the welfare of the adult with congenital heart disease. Copyright 2000 by W.B. Saunders Company PMID- 11486201 TI - Late outcome after the Fontan procedure. AB - Relatively little data exist on intermediate to late functional status and survival of patients after Fontan procedures. The first 500 patients undergoing a Fontan procedure at Children's Hospital (Boston, MA) were reviewed and then followed. There were 31 late failures among 410 patients contacted, and probability of survival at 10 years was 71.4%. Most patients were in functional class I or II, and only rare patients developed protein-losing enteropathy. Atrial flutter had developed in 16% of patients. Continued follow-up of Fontan patients will be necessary indefinitely. Copyright 1998 by W.B. Saunders Company PMID- 11486202 TI - Fenestration of the Fontan baffle: Benefits and complications. AB - Using a baffle fenestration or adjustable atrial septal defect in a modified Fontan operation is now a common practice. Hemodynamic benefits include increased cardiac index and systemic oxygen transport, as well as lower systemic venous pressure. The incidence and duration of pleural effusions is also reduced by this approach. Potential complications include those associated with the closure mechanism (snare or umbrella) as well as the possibility of paradoxical embolism. This review addresses our current understanding of the risks and benefits of this procedure. Copyright 1998 by W.B. Saunders Company PMID- 11486203 TI - Management of protein-losing enteropathy after the Fontan procedure. AB - Enteric loss of protein with the sequelae of edema, immunodeficiency, and hypercoagulability is being diagnosed with increasing frequency after Fontan procedure. The precise pathophysiological mechanism is unknown; however, protein losing enteropathy (PLE) after Fontan procedure is likely related to a hemodynamic derangement that is not easily detectable via standard hemodynamic measures presently obtainable in the cardiac catheterization laboratory. Treatment options include (1) symptomatic relief via diuretics, supplemental albumin infusion, and dietary change to high protein/high medium-chain triglyceride intake, (2) hemodynamic improvement via afterload reduction (angiotensin-converting enzyme inhibitors), repair of branch pulmonary artery stenoses, or coil embolization of aortopulmonary collaterals, (3) intestinal cell membrane stabilization via high-dose steroids or heparin infusion, and (4) attempt at alteration of the primary hemodynamic derangement via fenestration of the systemic venous baffle or via heart transplantation. Further understanding of the cause of PLE after Fontan procedure is needed before more effective treatment options can be used. Copyright 1998 by W.B. Saunders Company PMID- 11486204 TI - Conversion of the atriopulmonary Fontan connection to a total cavopulmonary connection. AB - Right atrial-to-pulmonary artery anastomosis (modified Fontan-Kreutzer operation) often results in substantial enlargement of the right atrium over time. Massive right atrial dilatation can cause loss of laminar flow, stasis, compression of the right pulmonary veins posteriorly, and leftward shift of the atrial septum. Some patients may develop arrhythmias related to atrial distension and multiple suture lines. Chronic elevation of the central venous pressure may cause liver dysfunction and/or protein-losing enteropathy in some patients. Some patients with atriopulmonary Fontan connections who develop a decline in functional status may benefit from conversion to a total cavopulmonary connection. Patients with significant ventricular dysfunction can usually only be treated by cardiac transplantation. However, other problems may be successfully managed by revision of the atriopulmonary Fontan connection to a total cavopulmonary anastomosis. Conversion to a total cavopulmonary connection should improve cardiac output by restoring laminar flow, by relieving compression of the right pulmonary veins, and by alleviating the leftward displacement of the atrial septum. Copyright 1998 by W.B. Saunders Company PMID- 11486205 TI - The extracardiac Fontan procedure. AB - The extracardiac modification of the Fontan procedure for complex single ventricle congenital cardiac disease is the latest attempt at improving on both the short-term and long-term results of this operation. This procedure differs from other modifications in that the inferior vena cava is detached from the heart and connected via a conduit to the pulmonary artery. It uses the fluid dynamic advantages of the cavopulmonary connection type of Fontan procedure but has other potential advantages as well. It is not necessary to cross-clamp the aorta to perform this procedure. Although quite a bit of prosthetic material is necessary to make the appropriate connections, none of this is exposed to the systemic circulation. Atrial flutter, a common early and late complication of other modifications of the Fontan procedure, appears to be less common with the extracardiac technique. The major disadvantages of this modification are the greater exposure of the venous pathway to potentially thrombogenic surfaces and the larger amount of prosthetic material involved in constructing the connection of the inferior vena cava to the pulmonary artery. Although the intermediate-term results with this procedure have been very favorable, issues that remain unresolved include the growth potential of the pathway, risk of late obstruction, and risk of atrial arrhythmias occurring late. Copyright 1998 by W.B. Saunders Company PMID- 11486206 TI - Sinus node dysfunction and atrial tachycardia after the Fontan procedure: The scope of the problem. AB - Similar to other atrial baffling procedures, the Fontan procedure exposes patients to ongoing morbidity and mortality. The development of the bradycardia tachycardia syndrome can have adverse effects on already-marginal hemodynamics and ventricular function. Patients with Fontan physiology and sinus node dysfunction can be managed with antibradycardic pacemakers. Atrial arrhythmias after "completion Fontan" are difficult to treat and usually require either antiarrhythmic agents, antitachycardic pacemakers, or radiofrequency catheter ablation of the re-entrant circuit. Successful treatment of atrial flutter occurs in only 50% to 70% of patients. There is a high recurrence rate of atrial flutter with any of the accepted management strategies. Copyright 1998 by W.B. Saunders Company PMID- 11486207 TI - Surgical variations and flow dynamics in cavopulmonary connections: A historical review. AB - Cavopulmonary connections (CPC) have been extensively used in the palliation of complex forms of congenital heart disease requiring some form of right heart bypass. The concept has evolved from unilateral CPC to total CPC (TCPC) in an attempt to prolong the palliated state, but the physiological implications remain only partially understood. We summarize some of the modifications and experimental data and their impact on flow distribution after the Fontan procedure. Copyright 1998 by W.B. Saunders Company PMID- 11486208 TI - Video-assisted thoracic surgical procedures in children. AB - The general principles and current applications of pediatric video-assisted cardiothoracic surgery (PVACTS) are reviewed. The purpose of PVACTS is to improve surgical quality and precision in selected operations. In the 1990s PVACTS has expanded to include the management of a variety of pulmonary, mediastinal, and cardiac lesions. Currently, PVACTS is carried out using a video camera connected to a low-profile scope and a specialized set of surgical instruments. PVACTS is an accepted modality for the diagnosis (by biopsy) of pleuropulmonary and mediastinal disease, and the treatment of pediatric empyema, spontaneous pneumothorax, and mediastinal cysts. Diaphragmatic plication, repair of chylous leak, and ligation of collateral vessels have all been done using PVACTS. PVACTS patent ductus arteriosus (PDA) ligation and vascular ring repair are being successfully carried out in several institutions. The technique at The Children's Hospital of Philadelphia is described. Indications and techniques for PVACTS lobectomy and pneumonectomy are less well established. Suggested anecdotal methods are described. Cardioscopy carries the hope of improving intracardiac repair, and has been applied to several lesions. The future of PVACTS depends on the surgeon's willingness to master it, industry's willingness to customize instruments for pediatric use, and developments in the fields of virtual imaging and augmented reality. Copyright 1998 by W.B. Saunders Company PMID- 11486209 TI - Minimal-access surgery for congenital heart defects. AB - Improved results with lower operative mortality and morbidity for corrective surgery for many congenital cardiac defects has stimulated a renewed interest in the use of surgical approaches other than a full midline sternotomy. In an effort to decrease pain and discomfort, shorten the recovery period, and improve the cosmetic result, several alternative approaches have been proposed and implemented, with varying results. Anterior thoracotomy in the inframammary area has been the most widely used incision and is most applicable to females patients past puberty, in whom the extent of breast tissue can be assessed more accurately. Complications with this approach including phrenic nerve injury and breast and chest wall deformities have been reported, although most reports describe satisfactory cosmetic results. We have used a midline approach limiting the incision over the xyphoid process either without a sternal incision (infants) or with division of the lower segment (patients younger than 3 to 4 years) with cephalad retraction to expose the heart and great vessels. From May 1996 to June 1997, 54 children had repair of a secundum-type atrial septal defect using a transxyphoid or ministernotomy approach. In 29, arterial cannulation was performed through the ascending aorta, and in 25 via the femoral artery. There were no instances in which conversion to full sternotomy was required, and complete repair with comparable ischemic and bypass time to full sternotomy was achieved in all patients. We have also used the same technique for repair of other congenital cardiac lesions, including ventricular septal defect and partial or complete atrioventricular canal defects, and in selected infants with tetralogy of Fallot. With this approach, cardioplegia for myocardial protection and left ventricular venting to prevent distention and to remove air from the heart can be used routinely. The adaptability of this technique to various cardiac defects and the ability to extend the incision if necessary make it an attractive alternative to other approaches for minimal-access cardiac surgery for congenital defects. Copyright 1998 by W.B. Saunders Company PMID- 11486210 TI - Use of modified ultrafiltration after repair of congenital heart defects. AB - The use of cardiopulmonary bypass (CPB) for repair of congenital heart defects exposes children to extremes of hemodilution and hypothermia. Exposure of blood to the foreign surfaces of the oxygenator and bypass circuit initiates a systemic inflammatory response. Adverse effects of CPB include increased capillary permeability and increased total body water (TBW), which often results in tissue edema and multiple organ dysfunction. A variety of techniques have been developed for reversing tissue edema and hemodilution after CPB, including ultrafiltration during CPB, postoperative peritoneal dialysis, postoperative continuous arterial venous hemofiltration, and aggressive use of diuretics. A technique termed modified ultrafiltration (MUF) has been developed at the Hospital for Sick Children in London. Unlike conventional ultrafiltration, MUF is performed in the immediate post-CPB period and removes excess water from the patient as well as provides a method of salvaging blood from the circuit. MUF has been shown to modulate the inflammatory response to CPB by removing inflammatory mediators including interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF). A prospective randomized trial of MUF showed improved hemodynamics with a reduction in TBW and decreased need for blood transfusion when compared with nonfiltered controls. MUF has been shown to improve left ventricular systolic function after CPB, resulting in increased systolic blood pressure and cardiac index. In a recent study, use of MUF significantly reduced the incidence of pleural effusions after cavopulmonary connection and the Fontan procedure. MUF is a useful adjunct to CPB in children and significantly decreases perioperative morbidity. Copyright 1998 by W.B. Saunders Company PMID- 11486211 TI - Management of postoperative arrhythmias and junctional ectopic tachycardia. AB - The critical postoperative period in children who have undergone palliative or corrective surgery for congenital heart disease is often complicated with cardiac arrhythmias. In spite of improved myocardial preservation techniques and better understanding of the surgical anatomy of the conduction system, postoperative arrhythmias are inevitable. Although most arrhythmias are transient, they can be associated with high mortality and morbidity if vigorous appropriate management is not instituted. This is especially true for postoperative junctional ectopic tachycardia. Lack of atrioventricular synchrony and reduced diastolic time secondary to a fast heart rate lead to decreased cardiac output, and the patient's condition tends to deteriorate rather rapidly. Recent advances in the management of postoperative arrhythmias have been gratifying. The most encouraging response has been that of junctional ectopic tachycardia to intravenous amiodarone. Also, the practice of routinely placing temporary epicardial wires in all patients undergoing surgery for congenital heart disease has provided a very useful diagnostic and therapeutic tool. Early recognition, precise diagnosis, and timely therapy can be very effective in avoiding adverse hemodynamic consequences of postoperative arrhythmias. Copyright 1998 by W.B. Saunders Company PMID- 11486212 TI - Early extubation of neonates and young infants after cardiac surgery. AB - Surgical intervention for many patients born with congenital heart disease has become routine in the first few months of life. Early extubation after cardiac surgery in neonates and young infants is feasible for many of these patients. This approach simplifies postoperative management, eliminates some of the complications related to postoperative care, shortens intensive care and hospital stays, reduces cost, and provides for a more efficient and positive convalescence for these young patients and their families. Management protocols for early extubation are described, and the outcomes reviewed. Copyright 1998 by W.B. Saunders Company PMID- 11486213 TI - Postoperative management after the Norwood procedure. AB - Staged reconstruction has become the preferred approach to hypoplastic left heart syndrome at many centers in the United States. The overall results of this strategy are most adversely affected by a high mortality at the initial stage, the Norwood procedure. The hemodynamic instability of a single ventricle providing blood flow in parallel to the systemic and pulmonary circulations combined with the stresses of cardiopulmonary bypass and circulatory arrest result in a precarious postoperative condition. Diligent perioperative management at this stage is essential to survival. To help simplify the complexity of single ventricle physiology, this article describes a mathematical model that identifies the key elements that affect systemic oxygen delivery. The importance of balancing the circulation is underscored. The value of monitoring both systemic arterial and venous oxygen saturations to assess systemic-to-pulmonary blood flow ratio is derived from this mathematical model and confirmed experimentally and clinically. Recent research using animal models of single-ventricle physiology is also described. Using these concepts and information, techniques for achieving adequate systemic oxygen delivery are discussed. Copyright 1998 by W.B. Saunders Company PMID- 11486214 TI - Reoperation after repair of atrioventricular canal defects. AB - Late events requiring reoperation after atrioventricular canal defect are most commonly related to failure of the repair of the mitral component of the common atrioventricular valve. The need for reoperation also occurs with new developments in the left ventricular outflow tract in the form of subaortic stenosis, particularly in patients with partial atrioventricular canal. Reoperations for other reasons in the population of operated patients with atrioventricular canal defects are rare events. Copyright 1998 by W.B. Saunders Company PMID- 11486215 TI - Long-term results of arterial switch repair of transposition of the great vessels. AB - The arterial switch operation has become the preferred surgical procedure for transposition of the great arteries worldwide. The low operative mortality at "low-risk" institutions has been well documented. The advantages of the arterial switch compared with atrial-level repairs include a lower incidence of arrhythmias and the likelihood of normal systemic ventricular function over the long term. However, the long-term sequelae of this operation must be continually evaluated, including the fate of the supravalvular pulmonary and aortic anastomoses, growth of the aortic root, competency of the neoaortic valve, patency of the coronary arteries, effects on the conduction system, and adequacy of ventricular function. These anatomic results, as well as the neurodevelopmental outcomes of these patients, are summarized in this review. Copyright 1998 by W.B. Saunders Company PMID- 11486216 TI - Late results of repair of truncus arteriosus. AB - Over the last three decades, management of truncus arteriosus has evolved with improving outcomes. Surgical repair is currently performed primarily during the neonatal period. This has prevented the severe morbidity and mortality resulting from congestive heart failure and pulmonary vascular obstructive disease. Although it has been 30 years since successful surgical repair of this lesion, the long-term follow-up studies have been limited to children who underwent truncus repair beyond the infancy period. In this article, we review the literature and also summarize the long-term results of truncus arteriosus at the University of California, San Francisco, where a repair in early infancy has been routine since 1975. A retrospective review was performed to assess long-term outcomes among 165 patients (81% of patients were < 1 year of age) who survived the initial hospital stay following complete repair of truncus arteriosus since 1975. There have been 23 late deaths, eight of which occurred within 6 months of repair and 13 of which occurred within 1 year. Ten of the late deaths were related to reoperations. The actuarial survival rate among all hospital survivors was 90% at 5 years, 85% at 10 years, and 83% at 15 years, and was essentially identical for infants alone. Significant independent risk factors for poorer long term survival were truncus with moderate to severe truncal valve insufficiency before repair. During the follow-up period, 107 patients underwent 133 conduit reoperations at a median of 5.5 years after the initial repair. In addition, 26 patients underwent 30 truncal valve replacements. Actuarial freedom from truncal valve replacement was 63% at 10 years among patients with truncal insufficiency before initial repair. Ten- to 20-year survival and functional status were excellent among infants undergoing complete repair of the truncus arteriosus. Copyright 1998 by W.B. Saunders Company PMID- 11486217 TI - Aortic insufficiency: Indications for surgery in children. AB - The goals of surgery in children with chronic aortic insufficiency are to prevent irreversible left ventricular dysfunction and to provide for long-term survival. In the past, surgical options included placement of a mechanical valve, a porcine bioprosthesis, or an aortic valve homograft. Complications from these options include thromboembolism, prosthetic valve endocarditis, limited durability, and lack of growth potential. The increasing utilization of the Ross procedure to treat chronic aortic insufficiency has led to new interest in the question of when to operate on a regurgitant aortic valve. This review focuses on the pathophysiology of aortic insufficiency and the invasive and noninvasive preoperative indices that may indicate the optimal time for aortic valve surgery in the pediatric population. Copyright 1998 by W.B. Saunders Company PMID- 11486218 TI - Anatomy of the aortic and pulmonary roots. AB - The use of the pulmonary autograft for aortic root replacement has renewed interest in the morphology of the arterial roots. In this article the basic construction of the roots, their anchorage, and their support from surrounding structures are reviewed. The arterial roots manifest a complex anatomy, with an intricate relation between the anulus and its adjacent structures, which span the transition from ventricle to the great vessel. The pulmonary root is anchored over its entire circumference to the thin myocardium of the free-standing pulmonary infundibulum. The anchorage of the aortic root is more extensive, being partly inserted into the thick left ventricular and septal myocardium and partly continuous with fibrous structures such as the membranous septum and the mitral valve. The pulmonary root is supported only by a slight collar of myocardium. The aortic root is better encased, supported by the more pronounced bulging of ventricular myocardium as well as the adjacent atrial myocardium and atrial septum. When the pulmonary autograft is used for aortic root replacement it will obtain maximum support from the surrounding tissues by implanting the autograft as proximal as possible. Copyright 1998 by W.B. Saunders Company PMID- 11486219 TI - Technique of pulmonary autograft aortic valve replacement in children (the Ross procedure). AB - Pulmonary autograft replacement has become a popular technique for relieving left ventricular outflow tract obstruction in children with aortic valvular or subvalvular disease. The advantages of the autograft are the avoidance of prosthetic valves that require anticoagulation and the potential for growth of a viable autograft aortic valve. Although many techniques for implantation of the autograft in the aortic outflow tract have been described, the most commonly preferred technique is the use of the autograft as a root replacement with reimplantation of the coronary ostia. This technique provides the optimal geometry of the autograft commissural attachments and also permits adjustment of the aortic annulus to enlarge the subaortic region in patients with complex left ventricular outflow tract obstruction or to narrow the aortic annulus in patients with primary aortic regurgitation and annular dilatation. Use of the autograft technique in children and young adults has resulted in excellent relief of outflow tract obstruction with low operative morbidity and mortality, and the intermediate results have been encouraging. The variations in surgical technique that have evolved have the potential to decrease the risk of late redilatation and regurgitation of the autograft valve and may provide better longevity. Copyright 1998 by W.B. Saunders Company PMID- 11486220 TI - The molecular basis of congenital cardiac disease. AB - Today, congenital cardiovascular disease is virtually always amenable to corrective or palliative surgical interventions. However, the mechanisms causing developmental anomalies of the heart and vessels have remained obscure until recently. This review presents genetic defects causing the pediatric vasculopathies; Marfan's syndrome, inherited supravalvar aortic stenosis, and Williams' syndrome. A synopsis of known mutations causing human cardiomyopathies in nuclear genes encoding contractile proteins, cardiomyocyte structural proteins, and mitochondrial proteins essential for cardiac energy production is provided. The molecular genetic evidence implicating single gene mutations in the pathogenesis of conotruncal anomalies (the 22q11 monosomy or "cardiac defects, abnormal facies, thymic hypoplasia, cleft palate, and hypocalcemia with deletions on chromosome 22" [CATCH-22] syndrome), heterotaxy syndromes, trisomies and atrioventricular canal defects, and secundum atrial septal defects is presented. The consequences of these genetic causes for diagnostic evaluation and perioperative care are emphasized. Single gene defects are a common cause of congenital cardiac disease. Copyright 1998 by W.B. Saunders Company PMID- 11486221 TI - Introduction. PMID- 11486223 TI - Tricuspid valvuloplasty in hypoplastic left heart syndrome. AB - Tricuspid valve regurgitation is a common finding in patients with hypoplastic left heart syndrome undergoing staged surgical reconstruction and can result from either abnormal valve morphology or incomplete leaflet coaptation due to annular dilatation. Significant tricuspid insufficiency imposes an additional volume load on the right ventricle and may have an important effect on survival. The spectrum of tricuspid valve anatomy found in hypoplastic left heart syndrome and surgical techniques available for the repair of atrioventricular valves are discussed. Tricuspid valvuloplasty during either the hemi-Fontan or Fontan stages of reconstruction carries a high success rate and is associated with improved right ventricular function. Copyright 1999 by W.B. Saunders Company PMID- 11486222 TI - Valvular repair for atrioventricular regurgitation in complex anomalies in modified Fontan procedure with reference to a single ventricle associated with a common atrioventricular valve. AB - Between January 1985 and October 1998, 169 of 372 patients who underwent a modified Fontan procedure had atrioventricular valve regurgitation (AVVR) that ranged in degrees from 1 to 4. Concomitant repair for AVVR was performed in the majority of cases. All but 12 patients had the AV valve repaired, mainly by circular annuloplasty; none had valve replacement. Although mortality was significantly higher in the AVVR cases (21 of 169 [12%]) than in the cases without AVVR (eight of 203 [3.9%]; P <.007, chi-square), actuarial survival in the AVVR cases was 83% at 5 years, 81% at 10 years, and 73% at 12 years. The degree of AVVR before surgery was 1.62 +/- 0.73 on average; 82 cases had more than grade 2 regurgitation. There was a significant decrease to 0.54 +/- 0.61 (P <.0001) after surgery in long-term survivors. Cases with AVVR can be treated with reasonable risk provided proper repair of the valve is performed. Circular annuloplasty is a simple and uniformly effective method with which to control regurgitation, even for the common AV valve. Copyright 1999 by W.B. Saunders Company PMID- 11486224 TI - Ebstein's anomaly: Indications for repair and surgical technique. AB - Ebstein's anomaly of the tricuspid valve is characterized by a regurgitant tricuspid valve with a large anterior leaflet and septal and tricuspid leaflets displaced into the right ventricle. Associated anomalies are common, especially atrial septal communication and right ventricular outflow tract obstruction. Clinical presentation includes a spectrum from minimal hemodynamic derangement and mild exercise intolerance to severe cardiopulmonary abnormalities and grave illness. The natural history of the disease is extremely variable; the worst prognosis is in the youngest patients, who often have associated cardiac abnormalities. Operative procedures range from ligation of a patent ductus arteriosus to cardiac transplantation. Tricuspid valvuloplasty has been advanced by several investigators and can be considered in nearly all cases. Repairs are based on the creation of a monocuspid or bicuspid valve with reduction of the enlarged annulus. While tricuspid valve repair or replacement is indicated for symptomatic children and adults, the management of the symptomatic neonate is difficult. Patients are stratified according to clinical status and morphology, then treated with observation, valve repair, conversion to single ventricle, or transplantation. We review the anatomy and pathophysiology of Ebstein's anomaly, the indications for surgery, and the appropriate timing of surgical intervention. Current techniques of tricuspid valvuloplasty are described and unresolved issues are discussed, including the role for superior cavopulmonary anastomosis, the management of symptomatic infants, and the necessity of ventricular plication in valve repair. Copyright 1999 by W.B. Saunders Company PMID- 11486225 TI - Valves in development for autogenous tissue valve replacement. AB - Currently available valve and conduit artery substitutes have one or more significant disadvantages including limited durability, thrombogenicity, susceptibility to infection, and a lack of growth potential. Prior attempts to use autologous tissues in the construction of valve or arterial substitutes to overcome some of these limitations have not been successful. The use of tissue engineering techniques to construct valve and arterial substitutes from individual autologous cell lines and biodegradable polymer scaffolds are now under investigation in the laboratory, and the initial short term results in animals have been encouraging. These tissue engineering techniques offer the possibility of creating structures for replacement of valves and conduit arteries which are viable and have the capacity for self-repair and therefore greater durability. In addition, these structures should be non-thrombogenic and less susceptible to infection, and will have growth potential. Copyright 1999 by W.B. Saunders Company PMID- 11486226 TI - Early and late results of fresh autologous pericardial valved conduits. AB - The objective of this study was to evaluate the early and late results of an autologous pericardial valved conduit in the pulmonary circulation. Between 1983 and 1997, 86 autologous pericardial valved conduits were used to achieve venous ventricle-pulmonary artery continuity. The mean patient age at the time of implantation was 4.16 +/- 4.10 years (15 days to 24 years). All patients had two dimensional postoperative and yearly Doppler echocardiograms in which the valve function and the presence of distal, valvar, and proximal stenosis were evaluated. There were 13 early deaths (15%). Twenty-one patients (24.4%) showed trivial, 54 (62.8%) mild, nine (10.4%) moderate, and two (2.3%) severe pulmonary regurgitation in the early postoperative period. The 73 survivors were monitored from 1 to 15 years (mean, 6.5 years; median, 7.1 years). There were five late deaths. The mean conduit diameter at the time of implantation was 15.8 mm, increasing to 18.21 mm at last evaluation (P <.0001). There were nine reoperations, with only one conduit replacement. Freedom from reintervention at 5 and 10 years was 89% and 80%, respectively. In conclusion, autologous pericardial valved conduits provide good early and excellent long-term results that compare favorably with those of other conduits. Copyright 1999 by W.B. Saunders Company PMID- 11486227 TI - Pericardial and synthetic monocusp valves: Indication and results. AB - Construction of a monocusp valve is a simple, expeditious, and inexpensive method to prevent the known sequelae of pulmonary insufficiency in the immediate post operative period whenever a transannular patch is used to establish right ventricular-pulmonary artery continuity. A detailed description of monocusp materials, construction and indications as well as late results are included. Copyright 1999 by W.B. Saunders Company PMID- 11486228 TI - Pericardial leaflet extension for aortic valve repair: Techniques and late results. AB - A modified technique to repair aortic valve is described. The technique consists of tailored extension of the old valve leaflets with gluteraldehyde treated autologous pericardium. The technique has excellent intermediate results and is an alternative to valve replacement in pediatric patients. Copyright 1999 by W.B. Saunders Company PMID- 11486229 TI - Allograft mitral valve replacement. AB - Allograft valves have been used in cardiac valve replacement for over 40 years. Early failures of allograft mitral valve replacement (AMVR) were attributed to inadequate allograft preservation or unsuccessful implantation techniques. Due to an improved understanding of mitral valve function gained from experience in mitral valve reconstruction, improved imaging techniques for preoperative assessment, and improved methods of allograft preservation, there has been a resurgence of interest in AMVR. The use of bioprosthetic valves in children and young adults is characterized by reduced durability and early valve failure. With the use of mechanical valves, there is a significant long-term risk of morbidity and mortality resulting from thrombosis or hemorrhagic complications from anticoagulation. Additionally, anticoagulation is often difficult to manage in children. The advantages of using allograft mitral valves include the avoidance of anticoagulation and the preservation of the subvalvar apparatus and its role in ventricular function. At our institution, successful early results with AMVR have been obtained in eight children and young adults with previous atrioventricular septal defects, previous placement of mitral valve prostheses, rheumatic valvular disease, and bacterial endocarditis. Our early results and those of others are encouraging for the continued use of AMVR in children and young adults. Long-term follow-up will be necessary to assess the durability and function of these allograft mitral valves. Copyright 1999 by W.B. Saunders Company PMID- 11486230 TI - Mitral valve repair in children using Carpentier's techniques. AB - Controversy remains regarding whether valve repair is preferable to valve replacement in children suffering from rheumatic mitral valve disease. To answer this question, 130 children aged 3 to 15 years (mean age, 11.8 +/- 2.8 years) undergoing surgery between January 1992 and December 1997 using Carpentier's techniques were reviewed. There were 111 cases of rheumatic valve diseases (85%), 17 cases of congenital mitral valve malformations (13%), one case of Barlow's syndrome (1%), and one case of bacterial endocarditis (1%). Valve dysfunction was classified into three types according to Carpentier's classification. There were 14 type II (leaflet prolapse), 78 type III (restricted leaflet motion), and 38 associated type II (anterior leaflet) and III (posterior leaflet) cases. There was one early (in-hospital) death (0.7%); the remaining 129 children were examined every 6 months. At the last examination, 99.2% of the patients were still alive, 96.8% were free of reoperation, and 89% showed no sign of significant residual stenosis or insufficiency on echocardiography. There have been no thromboembolisms. We conclude that mitral valve repair using Carpentier's techniques is the preferred procedure in the surgical treatment of mitral valve incompetence in children, even in those with rheumatic valvular disease. Copyright 1999 by W.B. Saunders Company PMID- 11486231 TI - Alternative nonvalved techniques for repair of truncus arteriosus: Long-term results. AB - Primary early repair has been advocated as the preferred surgical approach to truncus arteriosus in infancy. This approach usually includes the reconstruction of the right ventricle outflow tract using a valved extracardiac conduit. However, the longevity of these conduits and the risk of their replacement have been a subject of major concern. When used in early infancy, these conduits require repeated replacement because of internal obstruction or because the patient has outgrown the conduit. Our group first described the possibility of correction without an extracardiac conduit in 1990, although it has been performed at our institution since 1987. The ideal candidates for this type of correction include patients up to 5 months of age who have truncus arteriosus type I or II (Colllet-Edwards classification), no anomalous trajectory of the coronary artery crossing anteriorly to the truncus, and no significant pulmonary vascular resistance (pulmonary vascular resistance <7 Wood units). This surgical technique is described and the outcomes reviewed. Copyright 1999 by W.B. Saunders Company PMID- 11486232 TI - Late results of extracardiac Fontan repair. AB - The Fontan procedure by means of an extracardiac conduit was initially proposed for patients presenting with anomalies of intra-atrial anatomy. We extended this technique to all patients with functional anatomic single ventricle. Between 1988 and 1998, 206 patients with complex cardiacanomalies underwent a total extracardiac cavopuolmonary connection. In 202 patients we used a conduit, in 4 patients we performed an IVC to pulmonary artery direct anastomosis. Ten patients underwent Conversion of a filing atriopulmonary Fontan procedure to a total extracardiac cavopulmonary connection. Early deaths occurred in 10% of patients and the extracardiac conduit was taken down in 3 additional patients. The cause of death was myocardial failure in 13 patients. Pulmonary distortion or hypoplasia in 6. No deaths have occurred in our last 45 patients. We observed no cases of conduit obstruction and thrombosis. Arrthymias were present in 16 patients. Ten patients underwent conversion of a formed modified Fontan. There were no immediate postoperative deaths. These results demonstrate that the total extracardiac cavopulmonary connection provide good early and mildterm results and is the technique of choice for a Fontan type repair. Copyright 1999 by W.B. Saunders Company PMID- 11486233 TI - The favorable impact of arrhythmia surgery on total cavopulmonary artery Fontan conversion. AB - Patients who have undergone atriopulmonary Fontan procedures are at risk for significant atrial arrhythmias and obstructive connections, which have been related to intra-atrial suture lines, atrial enlargement, and somatic growth. Twenty patients (mean age, 17.3 +/- 6.8 years) had conversion to total cavopulmonary artery connection 8.9 +/- 2.1 years after the previous Fontan procedure (for atrial arrhythmias in 19 patients and for obstructive lesions in one). Arrhythmia ablative surgery evolved over the study period from "arrhythmia circuit cryoablation" (cryoablation lesions completing lines of block) to the more standard approaches of modified right-sided Maze and Maze-Cox III procedures. Preoperative functional New York Heart Association class was IV in nine patients, III in nine, and II in two. All patients survived. Two patients had prolonged chest drainage (there was pericardial effusion in one). The average length of hospital stay was 11.3 +/- 5.4 days; chest tubes were removed on day 8.5 +/- 5.4. There were no long-term deaths (mean follow-up period, 20.3 +/- 14.9 months; range, 2 months to 4 years). Late postoperative arrhythmias occurred in two patients who are receiving long-term antiarrhythmic medications. All patients have improved to New York Heart Association class I or II. Total cavopulmonary artery Fontan conversion in association with modified right-sided Maze or Maze Cox III procedures and pacemaker placement can be accomplished with low morbidity and mortality, and results in functional class improvement and control of life threatening arrhythmias. Copyright 1999 by W.B. Saunders Company PMID- 11486234 TI - Pulsatile pediatric ventricular assist devices: Current results for bridge to transplantation. AB - While pulsatile ventricular assist devices have gained widespread use in adult patients awaiting heart transplantation, only very limited experience with these devices exists in the pediatric population. In the past mostly non-pulsatile systems such as ECMO have been used to support pediatric patients with heart failure for very limited periods of time. Only recently have miniaturized pulsatile devices became available. This article describes the technical characteristics of these devices which have been implanted at the Deutsches Herzzentrum Berlin since 1992. The Berlin Heart assist device has since been used in 34 children between the ages of 6 days and 16 years. We report our strategy in patient selection, perioperative care and subsequent heart transplantation. The current literature on mechanical circulatory support in children is reviewed. Copyright 1999 by W.B. Saunders Company PMID- 11486235 TI - Surgical management of congenital and acquired pulmonary vein stenosis. AB - Pulmonary vein stenosis describes a variety of entities with a variable clinical course. The basic pathologic process appears to be fibrous intimal thickening, which can gradually cause obliteration of the lumen of the pulmonary veins at the atrial junction. In its most severe form, congenital pulmonary vein stenosis is a progressive disease with rapid pulmonary hypertension and rare survival beyond the first year of life. Surgical intervention has not been successful in this group. Other forms of pulmonary vein stenosis, including unilateral types associated with congenital heart disease, may be managed by various surgical treatments or pneumonectomy in some cases. Although multiple surgical techniques have been described for repair of congenital vein stenosis, the "sutureless" techniques recently described may have a place in the surgical treatment of this condition. In the most severe forms, lung transplantation appears to be an important option that should be considered early in the course of the disease. Copyright 1999 by W.B. Saunders Company PMID- 11486236 TI - The small left ventricle: How small is too small for biventricular repair? AB - Left ventricular hypoplasia is associated with a variety of congenital heart defects, including critical aortic stenosis, unbalanced atrioventricular canal, and total anomalous pulmonary venous connection, and is almost uniformly fatal without surgical or catheter-directed intervention. Accurately determining whether the left ventricle can adequately support the systemic circulation can be challenging and may be approached in a variety of ways, depending on the cardiac defect. The decision is more difficult in the present era of pediatric cardiology and cardiothoracic surgery because other options, such as the Norwood procedure and cardiac transplantation, are available to infants with left ventricular hypoplasia with improving survival. This report is a review of the present understanding of left ventricular hypoplasia and gives suggestions about how to stratify these complex patients to single versus two-ventricle repair. Copyright 1999 by W.B. Saunders Company PMID- 11486237 TI - An attempt at cloning I1 imidazoline receptors. PMID- 11486238 TI - Evaluation of cardiovascular parameters of a selenium-based antihypertensive using pulsed Doppler ultrasound. AB - The pharmacology of selenium is of much interest because selenium deficiency has been linked to cardiovascular diseases, cancer, and arthritis, and selenoenzymes are critical cellular antioxidants. We have previously reported that phenyl-2 aminoethylselenide (PAESe) and its derivatives represent a novel class of selenium-based antihypertensive agents that exhibit unique biochemical and pharmacologic properties. We now report on experiments designed to probe the hemodynamic mechanism of action of these compounds in spontaneously hypertensive rats (SHR). A noninvasive pulsed Doppler ultrasound probe was used to measure peak blood flow velocity in the aortic arch from the right second intercostal space. PAESe was found to increase peak aortic blood flow velocity (+44%), heart rate (+16%), and blood flow acceleration (+105%), while decreasing left ventricular ejection time (LVET) (-37%) concomitant with a decrease in mean arterial pressure (-54%). These results were compared with the known vasodilator hydralazine, which had similar effects on mean arterial pressure (MAP) and peak velocity but caused an increase in LVET (+42%) and a decrease in heart rate ( 18%). Taken together, our results suggest that PAESe decreases blood pressure via a decrease in peripheral resistance, which overcomes the initial increase in heart rate and acceleration to give a net decrease in MAP. PMID- 11486239 TI - Role of Ca2+-sensitive protein kinase C in phenylephrine enhancement of Ca2+ sensitivity in rat tail artery. AB - We investigated the role of protein kinase C (PKC) isoforms on changes in sensitivity of contractile mechanisms to intracellular Ca(2+) (force /[Ca(2+)]i) by phenylephrine (0.1-100 microM) in rat tail arterial helical strips using simultaneous measurements of force and [Ca(2+)]i. Force/[Ca(2+)]Ii induced by phenylephrine was greater than that induced by 80 mM K+. Force/[Ca(2+)]i induced by phenylephrine in physiologic saline solution or low Ca(2+) solution was dependent on the agonist concentration. Removal of Ca(2+) completely abolished the phenylephrine-induced contraction. The PKC inhibitors staurosporine and calphostin C inhibited the increase in force/[Ca(2+)]i induced by phenylephrine to a much greater extent than that induced by 80 mM K+. LY379196, a specific PKCbeta inhibitor, did not inhibit the increase of calcium sensitivity due to phenylephrine. The classic PKC isoforms, alpha, betaI, and II not gamma were demonstrated in the artery by immunohistochemistry. These results suggest that in rat tail arterial smooth muscle, PKCalpha, and not beta or gamma, mediates the increase of changes in sensitivity of contractile mechanisms to intracellular Ca(2+) to high dose of alpha1 receptor stimulation (phenylephrine 100 microM) on nonphysiologic conditions. PMID- 11486240 TI - A randomized, double-blind comparison of the efficacy and tolerability of once daily modified-release diltiazem capsules with once-daily amlodipine tablets in patients with stable angina. AB - A randomized, double-blind, parallel-group study comparing the efficacy and tolerability of once-daily diltiazem capsules with amlodipine tablets in patients with stable angina. After a run-in period of 1 to 3 weeks, 34 patients received once-daily diltiazem and 33 patients received amlodipine. Patients received either diltiazem, 240 mg/day, or amlodipine, 5 mg/day, for 2 weeks followed by diltiazem, 360 mg/day, or amlodipine, 10 mg/day, for 2 weeks. Standard treadmill exercise testing was the primary efficacy assessment. Patients also recorded incidence of angina attacks and use of glyceryl trinitrate spray. Both treatments gave significant improvement in time to onset of angina and time to maximal exercise. With the exception of amlodipine, 5 mg/day, both treatments gave significant increases in time to 1-mm ST segment depression. Diltiazem, 360 mg/day, gave a significant decrease in rate pressure product. There were no significant treatment differences in any of the exercise test parameters. Both treatments reduced incidence of angina attacks and use of glyceryl trinitrate spray. The incidence of edema was significantly less in patients receiving diltiazem. In conclusion, both treatments were effective in controlling patients' angina, but diltiazem was better tolerated, with a lower incidence of edema. PMID- 11486241 TI - Systemic uric acid administration increases serum antioxidant capacity in healthy volunteers. AB - Oxidative stress plays an important role in the development of atherosclerosis and contributes to tissue damage that occurs as a consequence, particularly in myocardial infarction and acute stroke. Antioxidant properties of uric acid have long been recognized and, as a result of its comparatively high serum concentrations, it is the most abundant scavenger of free radicals in humans. Elevation of serum uric acid concentration occurs as a physiologic response to increased oxidative stress-for example, during acute exercise-thus providing a counter-regulatory increase in antioxidant defenses. In view of its antioxidant properties, uric acid may have potentially important and beneficial effects within the cardiovascular system. We wished to investigate whether administration of uric acid was feasible and if it could have an impact on antioxidant function in vivo. We have, therefore, performed a randomized, placebo-controlled double blind study of the effects of systemic administration of uric acid, 1,000 mg, in healthy volunteers, compared with vitamin C, 1,000 mg. We observed a significant increase in serum free-radical scavenging capacity from baseline during uric acid and vitamin C infusion, using two methodologically distinct antioxidant assays. The effect of uric acid was substantially greater than that of vitamin C. PMID- 11486242 TI - Anti-ischemic action of estrogen-progestogen continuous combined hormone replacement therapy in postmenopausal women with established angina pectoris: a randomized, placebo-controlled, double-blind, parallel-group trial. AB - The benefit of treating postmenopausal women with established cardiovascular disease with combined estrogen-progestogen hormone replacement therapy (HRT) is controversial. This study investigated the effect of treatment with estradiol and norethisterone acetate on exercise tolerance and on the frequency and severity of ischemic attacks in postmenopausal women with stable angina pectoris. A total of 74 Chinese women were recruited for this 16-week double-blind, placebo-controlled trial. They were randomly allocated into two groups; one group received placebo/placebo/placebo and the other group received placebo/estrogen progestogen/placebo. Estrogen-progestogen continuous combined HRT increased both time to 1-mm ST depression (99.1 s, p < 0.05) compared with a mean decrease of 22.9 s with placebo (p < 0.05), and total exercise duration also showed a significant increase (32.7 s, p < 0.05) after treatment compared with placebo (2.5 s, p < 0.05). In addition, the total number of ischemic events/24 h during ambulatory electrocardiographic monitoring decreased by 0.82 events after treatment (p < 0.05) compared with an increase in the placebo group (0.94), a highly significant difference (p = 0.006). These results suggest that the administration of this particular combined hormone replacement preparation may have a beneficial effect on myocardial ischemia in postmenopausal women with established coronary disease. PMID- 11486243 TI - Adenosine A1 receptor antagonist prolongs survival in the hypoxic rat. AB - The hypothesis that adenosine A1 receptor (A1AdoR) selective antagonism limits cardiac depression and prolongs survival during acute global hypoxia was tested in a postinsult treatment model using KW-3902 ([8-(noradamantan-3-yl)-1,3 dipropylxanthine]), an A1AdoR selective antagonist. Rats were anesthetized, paralyzed, then ventilated with 8% O2 (hypoxia). In protocol I, 5 min after hypoxia, rats were treated with saline, drug vehicle, or KW-3902 (0.1 mg/kg i.v.). In protocol II, KW-3902 treatment occurred 2.5, 5, or 7.5 min after hypoxia. In protocol I, after hypoxia, left ventricular contractility, heart rate, and systemic mean arterial blood pressure decreased rapidly in saline-and vehicle-treated groups. In contrast, KW-3902 significantly attenuated the decline in these variables. Survival time (the time from the commencement of hypoxia until death) was more prolonged with KW-3902 (109.5 +/- 9.1 min) than with saline (37.6 +/- 5.0 min) or vehicle (35.0 +/- 4.2 min) (p < 0.001). In protocol II, survival time increased from 29.2 +/- 5.5 min in the 7.5-min treatment group to 109.5 +/- 9.5 min (5-min group) and 245.9 +/- 26.1 min (2.5-min group; p < 0.001). KW-3902 prolongs survival in this model, presumably by antagonizing A1AdoR-mediated inhibition of cardiac function. Also, treatment efficacy is highly time dependent. PMID- 11486244 TI - Angiotensin AT1 receptor antagonist irbesartan decreases lesion size, chemokine expression, and macrophage accumulation in apolipoprotein E-deficient mice. AB - Recent data suggest that angiotensin II AT1 receptor antagonists may be beneficial in the treatment of atherosclerosis. To clarify how AT1 receptor antagonists reduce atherosclerosis, the effect of irbesartan on atherosclerotic lesion development was determined in low-fat, chow-fed apolipoprotein (Apo) E deficient mice. Irbesartan (50 mg/kg per day) strongly decreased lesion development after a 12-week treatment period (lesion size: irbesartan treated, 20,524 +/- 4,200 microm(2) vs. control, 99,600 +/- 14,500; 79.4% inhibition, p < 0.001). This effect was not due to an effect of irbesartan on lipoprotein levels because irbesartan slightly increased total cholesterol levels and decreased the ratio of Apo A-I relative to Apo B levels. Immunochemical analysis of the atherosclerotic lesions using the mac3 monoclonal antibody showed the presence of macrophages in the lesions of control mice, whereas sections from irbesartan treated animals only showed occasional labeling in the lesion area. These data suggest that irbesartan inhibits monocyte/macrophage influx into the vessel wall. Therefore, expression levels of monocyte chemoattractant protein-1 (MCP-1), as well as other chemokines involved in macrophage infiltration into the lesion area, were measured in the aortic sinus of control and irbesartan-treated animals. Irbesartan treatment strongly decreased MCP-1 mRNA levels as well as MCP 1 immunostaining in the lesion area. This effect of irbesartan on MCP-1 occurred without an effect on CCR2, the receptor of MCP-1. Expression of macrophage inflammatory protein (MIP)-1alpha, another CC chemokine expressed in atherosclerotic lesions, was also reduced after irbesartan treatment, without effect on CCR3 and CCR5, the receptors of MIP-1alpha. Concomitantly, the expression of the angiogenic chemokines KC and MIP-2, which are functionally related to interleukin-8, were downregulated, whereas their shared receptor CXCR2 was upregulated. These data suggest that inhibition of the inflammatory component of lesion progression plays an important role in the inhibitory effect of AT1 receptor antagonists on atherosclerotic lesion formation. PMID- 11486245 TI - K(ATP) channel opening during ischemia: effects on myocardial noradrenaline release and ventricular arrhythmias. AB - Cardioprotection by K(ATP) channel openers during ischemia is well documented although ill understood. Proarrhythmic effects may be an important drawback. K(ATP) channel modulation influences neurotransmitter release during ischemia in brain synaptosomes. Therefore, we studied the effects of K(ATP) channel modulation on myocardial noradrenaline release and arrhythmias in ischemic rabbit hearts. Isolated rabbit hearts were perfused according to Langendorff and stimulated. Local electrograms were recorded and K+-selective electrodes were inserted in the left ventricular free wall. Cromakalim (3 microM) or glibenclamide (3 microM) was added 20 min prior to induction of global ischemia. After 15, 20, or 30 min of ischemia, hearts were reperfused and noradrenaline content of the first 100 ml of reperfusate was measured. Cromakalim (n = 16) prevented the second rise of extracellular [K(+)] in accordance with its cardioprotective effect. Cromakalim significantly reduced noradrenaline release after 15 min (mean, 169 +/- SEM 97 pmol/gr dry weight vs. control 941 +/- 278; p < 0.05) and 20 min of ischemia (230 +/- 125 pmol/gr dry wt vs. control 1,460 +/- 433; p < 0.05), but after 30 min of ischemia, the difference in noradrenaline release was no longer significant (cromakalim 2,703 +/- 1,195 pmol/gr dry wt vs. control 5,413 +/- 1,310; p = 0.08). Ventricular fibrillation or ventricular tachycardia occurred in 10 of 13 control hearts (77%) (n = 19), in six of 10 glibenclamide-treated hearts (60%) (n = 15), and in six of 14 cromakalim-treated hearts (43%) (p = NS). Cromakalim significantly accelerated onset of ventricular tachycardia or fibrillation (mean +/- SEM onset after 12.5 +/- 1.6 min ischemia vs. control 16.2 +/- 0.7 min; p < 0.05). Noradrenaline release occurred only in cromakalim-treated hearts with early-onset arrhythmias whereas no noradrenaline release was observed in cromakalim-treated hearts without ventricular tachycardia or fibrillation. Our results show that activation of the K(ATP) channel by cromakalim during ischemia reduces myocardial noradrenaline release and postpones the onset of irreversible damage, contributing to the cardioprotective potential of K(ATP) openers during myocardial ischemia. PMID- 11486246 TI - Evidence for the involvement of central I1 imidazoline receptor in ethanol counteraction of clonidine hypotension in spontaneously hypertensive rats. AB - Our previous studies have shown that ethanol counteracts centrally mediated hypotensive responses to clonidine. In this study, we investigated the relative roles of central alpha2-adrenergic and I1 imidazoline receptors in the antagonistic ethanol-clonidine hemodynamic interaction. The effects of selective blockade of alpha2- or I1 receptor by 2-methoxyidazoxan and efaroxan, respectively, on the blood pressure and heart rate responses to clonidine and subsequent ethanol administration were evaluated in conscious spontaneously hypertensive rats. Intracisternal administration of clonidine (1.5 microg/kg) produced significant (30 mm Hg; p < 0.05) and sustained (at least 60 min) decreases in blood pressure and heart rate. Systemic ethanol (1 g/kg), administered 10 min after clonidine, counteracted the hypotensive response and restored blood pressure to the preclonidine levels. Treatment with 2 methoxyidazoxan (0.16 microg/kg, intracisternal) or efaroxan (0.45 microg/kg, intracisternal) produced similar attenuation of the hypotensive and bradycardic responses to clonidine. The ability of ethanol to counteract the hypotensive action of clonidine was significantly (p < 0.05) attenuated in rats pretreated with efaroxan. The pressor response to ethanol lasted only 10 min compared with at least 60 min in the absence of efaroxan. In contrast, ethanol counteraction of clonidine-evoked hypotension was not altered when alpha2-adrenoceptors were blocked by 2-methoxyidazoxan. These findings suggest that centrally mediated hypotensive and bradycardic effects of clonidine in conscious spontaneously hypertensive rats involve activation of both alpha2-adrenergic and I1 imidazoline receptors. Furthermore, the findings suggest the dependence of a fully expressed ethanol counteraction of the hypotensive action of clonidine on functional I1 receptor within the central nervous system. PMID- 11486247 TI - Arterial baroreflex deficit induced organ damage in sinoaortic denervated rats. AB - To verify the independent role of the arterial baroreceptor dysfunction involved in target-organ damage in hypertension, sinoaortic denervated (SAD) rats were used as a model of arterial baroreflex (ABR) deficit. SAD, isolated aortic denervated (AD), and isolated sinus-denervated (SD) rats were instrumented to record blood pressure (BP), heart rate (HR), BP variability (BPV), HR variability (HRV), ABR function control of heart period (ABR-HP), and BP (ABR-BP). Vascular maximum contractile/relaxant function was determined and organ damage was estimated by observation of morphologic changes. Short-term (postoperative 1 week) SAD caused hypertension and tachycardia in rats. Eighteen weeks after operation, BP and HR values in SAD and SD rats were not different from those in sham-operated rats, but AD rats were hypertensive compared with control group. Although 24-h mean BP values of long-term SAD rats were not different from those of sham-operated rats, 24-h BPV of SAD rats was significantly higher than that of sham-operated rats. Arterial baroreflex function in short-term SAD rats was significantly less than in sham-operated rats, whereas in long-term SAD rats, ABR HP and ABR-BP were higher than those in short-term SAD rats, but were still significantly lower than those in control groups. At postoperative 18 weeks, baroreflex function in SAD and AD rats was significantly less than function in SD and control groups. SBPmax after phenylephrine and DBPmin after nitroprusside were significantly higher in SAD, AD, and SD rats than in control rats. Baroreflex function was negatively correlated to DBPmin and SBPmax in all denervated rats (n = 44). Some morphologic changes were found 18 weeks after denervation in heart, kidney, and small artery in SAD, AD, and SD rats. Baroreflex function in all denervated rats was negatively related to 24-h BPV values. In contrast, 24-h BPV values in SAD, AD, and SD rats were positively related to organ-damage score. A negative correlation between ABR function and end-organ damage score was found. Arterial baroreflex deficit played an independent and important role in organ-damage in SAD rats with significantly elevated 24-h BPV. PMID- 11486248 TI - Three thiadiazinone derivatives, EMD 60417, EMD 66430, and EMD 66398, with class III antiarrhythmic activity but different electrophysiologic profiles. AB - The thiadiazinone derivatives EMD 60417, EMD 66430, and EMD 66398 were developed as class III antiarrhythmic agents. Their chemical structure is closely related to that of their calcium-sensitizing congener [+]-EMD 60263, and EMD 66398 possesses the methylsulfonylaminobenzoyl moiety present in the prototypical IKr blocker E-4031. We compared the electrophysiologic effects of these compounds with standard drugs (almokalant, E-4031, quinidine) in cardiac myocytes from guinea-pig ventricle and human atrium by whole-cell patch-clamp technique. The test compounds' class III action, which is related to impairment of K+ channel function, was confirmed by action potential measurements. EMD 60417, EMD 66430, EMD 66398, and almokalant (1 microM each) reversibly prolonged the action potential duration in guinea-pig myocytes. In the same cells, the rapidly activating component IKr of the delayed rectifier K+ current, which has been defined by its sensitivity to E-4031, was reduced by EMD 60417, EMD 66430, EMD 66398, and almokalant. Inhibition of IKr was concentration-dependent as determined by attenuation of tail currents. The slowly activating component IKs of the delayed rectifier K+ current was not affected. The inward rectifier K+ current IK1 was not influenced at potentials close to the reversal potential. Transient and sustained outward K+ currents (Ito, Iso) measured in human atrial myocytes were not altered by any EMD compound. L-type Ca2+ current was hardly affected at concentrations of 1-10 microM, but sodium current was decreased. Action potential prolongation by EMD 60417, EMD 66430, and EMD 66398 is due to block of IKr. INa is inhibited at higher concentrations by EMD 66430 and EMD 60417. EMD 66398 is more potent and selective for IKr than EMD 60417 and EMD 66430, and thus resembles E-4031 in structure and function. PMID- 11486249 TI - Effects of the L- and N-type calcium channel blocker cilnidipine on growth of vascular smooth muscle cells from spontaneously hypertensive rats. AB - Cultured vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) show exaggerated growth compared with cells from Wistar-Kyoto (WKY) rats. Calcium antagonists have recently been reported to have an in vivo antiproliferative effect on hypertensive cardiovascular organs. We investigated the effects of the calcium antagonist cilnidipine that blocks both L- and N-type calcium channels on the growth of VSMC from SHR. Cilnidipine (1 and 10 microM) significantly inhibited basal DNA synthesis in VSMC from both rat strains; the inhibition was significantly larger in VSMC from SHR than in cells from WKY rats, and was significantly greater than effects of nifedipine. Cilnidipine (1 microM) significantly inhibited serum-stimulated DNA synthesis in VSMC from both rat strains. The inhibition was more marked in VSMC from SHR than in cells from WKY rats. Angiotensin II, platelet-derived growth factor (PDGF)-AA, and phorbol-12 myristate-13-acetate dose-dependently increased DNA synthesis in VSMC from SHR but not in cells from WKY rats. Cilnidipine (1 microM) significantly suppressed this increase in DNA synthesis in VSMC from SHR. Expression of basic fibroblast growth factor (bFGF), transforming growth factor-beta1, and PDGF A-chain mRNAs was markedly greater in VSMC from SHR than in cells from WKY rats. Cilnidipine (1 microM) significantly inhibited the expression of TGF-beta1 mRNA in VSMC from SHR but not in cells from WKY rats. These findings suggest that cilnidipine exerts its antiproliferative effects through the inhibition of DNA synthesis induced by growth-promoting factors and by inhibiting the expression of TGF-beta1 mRNA in VSMC from SHR. PMID- 11486250 TI - Transient outward current inhibition by propafenone and 5-hydroxypropafenone in cultured neonatal rat ventricular myocytes. AB - The antiarrhythmic agent propafenone and its primary electropharmacologically active metabolite, 5-hydroxypropafenone, are known inhibitors of cardiac myocyte repolarizing currents. We recently documented potent propafenone inhibition of the transient outward potassium current (Ito) in human atrial myocytes from patients in the newborn and infant age range. In the current study we characterized ventricular Ito inhibition by propafenone and 5-hydroxypropafenone in neonatal myocytes enzymatically isolated from 2-day-old Sprague-Dawley rat pups. Using the whole-cell patch-clamp technique in ventricular myocytes kept in primary culture for 1-4 days, we observed comparably potent Ito inhibition by both agents, yielding 50% maximal inhibitory concentration (IC50) values of 2.1 +/- 0.5 and 1.5 +/- 0.2 microM for propafenone and 5-hydroxypropafenone, respectively. Ito blockade by both of these agents was time, concentration, and voltage dependent, but use independent. There was no drug effect on steady-state voltage dependence of Ito inactivation, or on the time course of Ito recovery from inactivation. These findings are consistent with an open channel-blocking mechanism as suggested by other models. We conclude that both propafenone and 5 hydroxypropafenone are potent Ito inhibitors in neonatal rat ventricular myocytes, with potencies exceeding those demonstrated for propafenone in adult rat ventricular myocytes or in human atrial myocytes from patients of all ages. PMID- 11486251 TI - Incremental importance of peak-exercise plasma levels of endothelin-1 and natriuretic peptides in chronic heart failure. AB - Chronic heart failure (CHF) studies investigating the clinical, hemodynamic, and therapeutic importance of endothelin-1 (ET-1), atrial natriuretic peptide (ANP), and brain natriuretic peptide (BNP) are largely based on resting plasma levels, which may vary with prior exertion and postprandial status. This study investigated the importance of peak-exercise plasma levels of ET-1, ANP, and BNP in the assessment of left ventricular (LV) systolic function. Thirty-six male patients ages 58 +/- 10 (mean +/- SD ) with NYHA class I-IV CHF due to coronary artery disease or idiopathic dilated cardiomyopathy were enrolled. LV systolic function was assessed by echocardiography and radionuclide ventriculography. Resting and peak cardiopulmonary exercise venous blood sampling and treadmill exercise testing were performed in the fasting state. Resting plasma levels of ET 1, ANP, and BNP were elevated compared with reference laboratory normal values. Exercise induced significant (p < 0.0001) increase in plasma levels of ET-1, ANP, and BNP. On univariate analysis peak-exercise plasma levels of ET-1, ANP, and BNP were more closely related to echocardiographically determined LV end-diastolic diameter and end-systolic diameter than their resting values. Multiple step-wise regression models identified resting and peak-exercise plasma levels of ET-1 and ANP but only the resting BNP as independent predictors of LV dimensions and systolic function. Peak exercise plasma levels of ANP and ET-1 are potentially more reliable and important than their resting levels as markers of LV systolic dysfunction and LV dimensions in patients with heart failure. PMID- 11486252 TI - Amp 579 reduces contracture and limits infarction in rabbit heart by activating adenosine A2 receptors. AB - To determine the mechanism by which AMP 579, an adenosine A1/A2 agonist, administered at reperfusion protects ischemic myocardium, buffer-perfused rabbit hearts were subjected to 30 min of global ischemia and 2 h of reperfusion. AMP 579 (500 nM) was included in the reperfusate for the first 70 min. Average left ventricular diastolic pressure during reperfusion in hearts receiving AMP 579 was lower than that in control hearts (17.9 +/- 2.4 vs. 39.0 +/- 6.5 mm Hg, p < 0.05), indicating attenuation of contracture. Left ventricular developed pressure and coronary flow during reperfusion were also significantly improved with AMP 579 treatment. AMP 579's anti-contracture effect was blocked by the adenosine A2 receptor antagonist 8-(3-chlorostyryl)caffeine (CSC), but not by the A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). CSC, but not DPCPX, also blocked AMP 579's ability to preserve developed pressure and coronary flow in these hearts. AMP 579 significantly reduced infarction in isolated hearts subjected to regional ischemia. The anti-infarct effect again was abolished by CSC but not by DPCPX. Finally, we tested whether 5'-(N-ethylcarboxamido)adenosine (NECA), another A1/A2 agonist, also administered for the initial 70 min of reperfusion, could duplicate the anti-infarct effect of AMP 579. One-hundred nanomolar NECA duplicated the protection, but neither 50 nM CGS21680, a selective A2 agonist, nor 100 microM adenosine was protective. Therefore, AMP 579 given at reperfusion reduces contracture and infarction. Anti-contracture and anti-infarct effects require the adenosine A2, but not the A1, receptor suggesting that prevention of contracture and tissue salvage are mechanistically related. Not all A2 agonists were able to duplicate the anti-infarct effect, suggesting something unique about AMP579. PMID- 11486253 TI - Irbesartan effects on renal function in patients with renal impairment and hypertension: a drug-withdrawal study. AB - The blood-pressure lowering activity, tolerability, and safety of irbesartan was evaluated in 52 hypertensive patients with chronic renal insufficiency. After a 3 week placebo period, once-daily irbesartan was administered for 12 weeks at a daily dose of 150 mg titrated to 300 mg. A second, non-angiotensin-converting enzyme inhibitor, antihypertensive drug was added after 8 weeks as needed. Twenty four-hour creatinine clearance was determined and renal clearance studies of inulin and para-aminohippurate were done in a subset of 11 patients. Trough sitting blood pressures were reduced at the end of the first week in all groups. At weeks 4, 8, and 12 the reductions in systolic blood pressure/diastolic blood pressure averaged -11.9/-8.7, -10.8/-9.4, and -14.7/-12.1 mm Hg in patients with mild renal insufficiency and -7.7/-6.3, -13.1/-11.8, and -14.1/-10.6 mm Hg in patients with moderate-to-severe renal insufficiency. Creatinine clearance, glomerular filtration rate, and effective renal plasma flow were stable. Irbesartan was withdrawn in only five patients because of adverse clinical or laboratory experience. Hyperkalemia (>6 mEq/l) requiring discontinuation of irbesartan occurred in only one patient. Once-daily irbesartan given as monotherapy at dose of 150-300 mg or in combination with other antihypertensive drugs is effective in reducing blood pressure in hypertensive patients with chronic renal disease. Irbesartan regimens are well tolerated in all groups. In addition, the blood pressure-lowering effect of irbesartan is accompanied by a significant reduction in proteinuria in patients with chronic renal insufficiency. PMID- 11486254 TI - Personal hygiene and life expectancy improvements since 1850: historic and epidemiologic associations. PMID- 11486255 TI - Preventing infectious disease in the domestic setting: a risk-based approach. PMID- 11486257 TI - Transmission of gastrointestinal diseases: hygiene as the final barrier. PMID- 11486256 TI - The correlation between sustainable development and home hygiene. AB - Hygiene refers to the science of the establishment and maintenance of (human) health. In everyday life, hygiene is closely associated with good housekeeping. This article will focus on home hygiene in relation to cleaning, on microorganisms, and on sustainable development of domestic technology. In domestic cleaning there are two beneficial effects: the aesthetic aspects of cleanness and the removal of microorganisms. In cleaning science substantial attention is paid to the interrelation between cleaning and removal of microorganisms. It appears that the parameters of the cleaning process and the detergent properties play a significant role in this interrelation. Changing technology to reduce the environmental impact of household cleaning not only influences the household activities and the functional performance of the cleaning processes but also has an impact on the level of hygiene. Results are presented of research in which the hygiene has been evaluated in relation to such changes. One option recently studied to reduce the environmental impact of households is the reduction of water consumption by water reuse. In such concepts water is used in successive steps for various processes before it is drained. The potential impact of such systems and of the use of rain water on the level of home hygiene is discussed. PMID- 11486258 TI - Pandora's box: old and new emerging infections in at-risk populations. PMID- 11486259 TI - Listeria and listeriosis: risk assessment as a new tool to unravel a multifaceted problem. PMID- 11486260 TI - Hygiene and health - the need for a holistic approach. AB - The holistic principles of hygiene and public health have contributed substantially to an increase in life expectancy by more than 30 years and in life quality since the beginning of the 20th century. Frank, Pettenkofer, Nightingale, Pasteur, Lister, and Koch have been pioneering protagonists of the holistic approach to hygiene and public health. Socioeconomic development and related factors such as nutrition status and food hygiene, housing conditions, water supply and sewage systems, and education (including motivation for personal hygiene) have obviously been of more importance for life expectancy and life quality than progress in curative medicine, such as availability of microbial diagnosis, vaccination, and antibiotics. Today, new risk factors for infectious diseases arise, even in developed countries. These risk factors arise from emerging pathogens, antibiotic-resistant microorganisms, changing demographic patterns, an increasing amount of ambulatory and home care, socioeconomic and environmental changes, technical environments, worldwide distribution of food, and changing human behavior with a decreased awareness of microbial threats. These new challenges worldwide make a renewal of the holistic approach of hygiene and public health both urgent and necessary. On the basis of historic experience, policies that focus on surveillance and control, diagnosis, and therapy only can be assumed to be both insufficient and inefficient in controlling the new challenges in infectious diseases. Experiences in Germany with a holistic hospital hygiene strategy since 1976 provide encouragement for the promotion of holistic health concepts. Risk assessment, risk management, and risk communication are basic steps of a modern holistic strategy. Hygiene has the potential to act as a moderator of diverging positions of different disciplines within this renewed approach. PMID- 11486261 TI - Hygiene and health: an epidemiologic link? PMID- 11486262 TI - The role of handwashing in improving hygiene and health in low-income countries. PMID- 11486263 TI - Infection control practices in institutional settings. PMID- 11486264 TI - Prevention and control of viral hospital infections. PMID- 11486265 TI - The potential benefits of infection control measures in the home. PMID- 11486266 TI - Biofilm formation: attachment, growth, and detachment of microbes from surfaces. PMID- 11486267 TI - Biofilms: their impact on health and their recalcitrance toward biocides. PMID- 11486268 TI - The future control of bacterial resistance to antimicrobial agents. PMID- 11486269 TI - Mechanisms of bacterial insusceptibility to biocides. AB - Bacterial insusceptibility to biocides is of two types, intrinsic and acquired. Intrinsic insusceptibility is a natural property of an organism and is shown by bacterial spores, mycobacteria, and gram-negative bacilli. Cellular impermeability is a major factor, and in some cases active efflux pumps play an important role. A special example is that of phenotypic (physiological) adaptation to intrinsic resistance found in bacteria present in biofilms. Acquired resistance arises through mutation or via the acquisition of plasmids or transposons; efflux of biocide is a major mechanism, although plasmid-mediated inactivation has also been shown to occur. An additional aspect that must be considered is the stringent response elicited in bacteria on exposure to inimical agencies. There is a possible linkage between certain biocides and antibiotic resistance under experimental conditions. PMID- 11486270 TI - Variation in administrators' and clinicians' attitudes toward critical elements of an infection control program and the role of the infection control practitioner in New South Wales, Australia. AB - BACKGROUND: Debate remains over the core activities of infection control (IC) programs. Differences in stakeholder opinions must be considered if consensus panel guidelines and recommendations are to be broadly applied. This article describes a survey of administrators and clinicians employed in hospitals in New South Wales, Australia. Respondents self-reported their levels of agreement with affirmative statements regarding the role of the infection control practitioner (ICP) and the essential requirements and infrastructure of IC programs. METHOD: The study population included administrators and clinicians in each public, private, and freestanding day hospital in New South Wales. Respondents reported the intensity of their agreement with 16 affirmative statements relating to IC program infrastructure and resources and the ICP's role and responsibilities. RESULTS: The overall response rate was 62.1% (587/945). Clinicians (349/587) and administrators (238/587) accounted for 59.5% and 40.5% of the response rate, respectively. Overall, administrators and clinicians reported greatest levels of agreement for those elements not requiring additional resources. CONCLUSION: The extent of divergence between administrators and clinicians is not so great that it can not be resolved. Our findings demonstrate the degree of administrator support that clinicians can expect for each element. We advocate better communication between clinicians and administrators in conjunction with objective strategic planning. Our findings provide a guide for ICPs to either establish or negotiate the core components of their IC program. PMID- 11486271 TI - Body piercing as a risk factor for viral hepatitis: an integrative research review. AB - The relationship of viral hepatitis and body piercing has been investigated in 12 research studies published between 1974 and 1997. Because there are often a large number of unknowns regarding the cause of viral hepatitis, most of the studies were conducted to identify multiple risk factors for hepatitis. Eight of the 12 studies identified percutaneous exposure, including body piercing and ear piercing, as a risk factor for viral hepatitis. Six studies found that hepatitis seropositivity was significantly associated with ear piercing. Conclusions indicate that evidence is sufficient to institute public health education along with regulation of the body-piercing industry. Recent research has found a significant increase in legislative efforts to regulate body piercing. Little research has focused directly on body piercing as a specific risk factor for viral hepatitis. Further research in this area is necessary to increase the understanding of hepatitis transmission by body piercing. PMID- 11486272 TI - Descriptive study of nosocomial infections in a short-term inpatient behavioral health setting. AB - BACKGROUND: Surveillance methods for nosocomial infections, with the use of a sound epidemiologic foundation, have not been published for behavioral healthcare settings. METHODS: Nosocomial infection rates in a behavioral health care setting were quantified by integrating the following: (1) rounds/review of selected charts, (2) microbiology reports, and (3) records of antimicrobials dispensed. Incidence density rates were calculated with 1000 patient-days as the denominator. RESULTS: The incidence of nosocomial infections of all types among all patients was 3.9 infections per 1000 patient-days. Rates among specific groups of behavioral patients ranged from children (6.4 infections per 1000 patient-days) to adults (3.9 infections per 1000 patient-days) to adolescents (3.2 infections per 1000 patient-days). Adult substance-abuse patients had a rate of 1.9 infections per 1000 patient-days. The most frequently diagnosed nosocomial infection among adult substance-abuse and behavioral patients was bronchitis at 0.5 and 0.6 infections per 1000 patient-days, respectively. The most frequently diagnosed nosocomial infection among adolescents and children was cellulitis at 0.6 and 1.3 infections per 1000 patient-days, respectively. Female patients experienced significantly more nosocomial infections than male patients, except among adult substance-abuse patients. Nosocomial infections usually occurred in patients who were hospitalized longer than the average length of stay, except among children. CONCLUSION: Nosocomial infections occur in behavioral health settings. Surveillance methods, with a sound epidemiologic foundation, must be developed. PMID- 11486273 TI - Structural organization of the human MS4A gene cluster on Chromosome 11q12. AB - CD20, the high-affinity IgE receptor beta chain (FcepsilonRIbeta), and HTm4 are structurally related cell surface proteins expressed by hematopoietic cells. Recently, 16 novel human and mouse genes were identified that encode new members of this nascent protein family that we have named the membrane-spanning 4A gene family, with at least 12 subgroups (MS4A1-MS4A12). In the current study, we identified three additional human MS4A genes: MS4A4E, MS4A6E, and MS4A10. All family members have at least four potential transmembrane domains and N- and C terminal cytoplasmic domains encoded by distinct exons, except MS4A6E which contains two transmembrane domains. Otherwise, the 12 currently identified MS4A genes share common structural features and similar intron/exon splice boundaries, and are clustered along an approximately 600-kb region of Chromosome 11q12. In contrast to other MS4A genes, MS4A4E, MS4A6E, and MS4A10 transcripts were rare and not detected among hematopoietic cells and most nonlymphoid tissues. Sequence polymorphisms were identified in the MS4A6E gene and common splice variants were observed for the MS4A4A, MS4A5, MS4A6A, and MS4A7 genes. Thus, the MS4A family currently includes 24 distinct human and mouse genes. Like CD20 and FcepsilonRIbeta, the 10 other human MS4A family members are likely to be components of oligomeric cell surface complexes involved in signal transduction in diverse cell lineages. PMID- 11486274 TI - Single-nucleotide polymorphism detection by denaturing high-performance liquid chromatography and direct sequencing in genes in the MHC class III region encoding novel cell surface molecules. AB - The class III region of the human major histocompatibility complex (MHC) contains approximately 59 genes, many of which encode polypeptides with a variety of different functions. Eight of these genes are of particular interest because they encode novel surface molecules that could be involved in immune and/or inflammatory responses and are excellent candidates as disease susceptibility loci. These molecules are members of two different superfamilies, the immunoglobulin superfamily (1C7, G6B, and G6F genes) and the leucocyte antigen-6 superfamily (G6C, G6D, G6E, G5C, and G5B genes). Some level of variation was found when overlapping genomic DNAs from different haplotypes were compared. The present work describes a systematic search for single-nucleotide polymorphisms (SNPs) in these genes using direct sequencing and denaturing high-performance liquid chromatography (DHPLC) in 24 unrelated healthy individuals. We validated the DHPLC methodology by first studying the 1C7 gene. This gene was directly sequenced in all 24 samples, and DHPLC was found to resolve all the polymorphic sites present in the heterozygote samples tested. We screened the rest of the genes by DHPLC only, and only those chromatograms that revealed a polymorphic profile were sequenced. We detected one SNP every 489 bp in the 18 kb of DNA studied, corresponding to theta = 4.61x10-4. The diversity in noncoding regions is 1 SNP/560 bp, but a higher frequency was detected in coding regions with 1 SNP/423 bp corresponding to theta =5.33x10-4. Of the coding SNPs, 63.6% caused amino acid substitutions. The power of this study is emphasized by the fact that of the 37 SNPs/indels detected, only 6 can be found in the SNP database at the NCBI. PMID- 11486276 TI - Identification of quantitative trait loci controlling activation of TRBV4 CD8+ T cells during murine gamma-herpesvirus-induced infectious mononucleosis. AB - The murine gamma-herpesvirus, MHV-68, shares important biological and genetic features with the human gamma-herpesvirus, Epstein-Barr virus. Following intranasal infection, mice develop an infectious mononucleosis-like syndrome accompanied by increased numbers of activated CD8+ T cells in the blood. A consistent feature of the CD8+ T-cell activation is a marked increase in the frequency of cells expressing a TRBV4+ T-cell receptor. Previous studies suggested that the magnitude of TRBV4 expansion varied significantly among mouse strains, and was influenced by both MHC and non-MHC genes. Detailed analysis of strains with high (C57BL/6) or low (DBA/2) TRBV4 CD8+ T-cell expansion showed that differences in the degree of expansion were not a consequence of variation in genetic susceptibility to the viral infection. Rather, the magnitude of the TRBV4 CD8+ T-cell expansion correlated with differences in expression of the unidentified stimulatory ligand on activated, latently infected B cells. In the present study, analysis of TRBV4 expansion in C57BL/6, DBA/2, B6D2 F1 mice, BXD recombinant inbred strains, and the progeny of C57BL/6xDBA/2 F1 hybrids backcrossed to C57BL/6 demonstrated strong cumulative dominance of the low DBA/2 trait and moderately high heritability (h2 approximately 0.5). Two quantitative trait loci (QTLs) strongly associated with variance in TRBV4 expansion were identified using simple and composite mapping procedures. The first QTL is located on Chromosome (Chr) 17, near or proximal to H2. The second QTL is located on Chr 6 in a region spanning the Tcrb and Cd8a loci. PMID- 11486275 TI - CD150 is a member of a family of genes that encode glycoproteins on the surface of hematopoietic cells. AB - Human CD150 (SLAM) is a glycoprotein expressed on the surface of T, B, natural killer, and dendritic cells. The extracellular domain of CD150 is the receptor for measles virus and CD150 acts as a co-activator on T and B cells. We characterized the mouse and human CD150 genes, each of which comprises seven exons spanning approximately 32 kb. Mouse CD150 mRNA was detected in T cells and in most thymocyte subsets, except CD4-8- cells. Surprisingly, the CD4-8- thymocytes of CD3gammadeltanull mice, but not of Ragnull or severe combined immunodeficiency mice, expressed CD150. Whereas high levels of CD150 were found in Th1 cells, only small amounts were detectable in Th2 cells. CD150 expression was up-regulated upon in vitro activation of mouse T cells by anti-CD3. The complete mouse CD150 gene is highly homologous to its human orthologue in terms of nucleotide sequences and intron/exon organization. The human genomic sequences indicate that all isoforms detected so far have arisen from alternative splicing events. As judged by fluorescence in situ hybridization, mouse CD150 mapped to Chromosome (Chr) 1, band 1H2.2-2.3, and human CD150 was found on Chr 1q22. Human and mouse CD150 share sequence homologies with six other genes, five of which - CD84, CD229 (Ly-9), CD244 (2B4), CD48, and 19A - are localized in a 250-kb segment in close proximity to the human gene. Their location and their sequence similarities strongly suggest that the CD150 family of cell surface receptors arose via successive duplications of a common ancestral gene. PMID- 11486277 TI - The cell surface-expressed HSC70-like molecule preferentially reacts with the rat T-cell receptor Vdelta6 family. AB - We previously showed that the cell surface-expressed Mr 70,000 heat shock cognate (hsc70, a constitutively expressed member of the hsp70 family) protein-like molecule (#067 molecule) interacts with rat CD3+, CD4-, CD8-, T-cell receptor (TCR)alphabeta-, natural killer recetor-P1- T cells. This 70hsc-like molecule was also suggested to present cellular peptide antigens to these T cells. In the present study, we identified the genetic structure of the TCR by establishing T cell hybridomas between these T cells and mouse BW5147 cells. Our data indicated that these T cells preferentially used TCRs with the Vdelta6 family. Analysis of the nucleotide sequence of the CDR3 junctional portion showed that there are substantial diversities, with insertion of seven to nine amino acid residues. These data provide indirect evidences for our hypothesis that an hsc70-like molecule could be presented together with cellular peptide antigens to particular T cells with TCR gammadelta chains. Since the expression of this hsc70-like #067 antigen on the cell surface is usually induced along with cell transformation by activated oncogenes, T cells with the TCR Vdelta6 family are likely to contribute to host resistance to tumor cells. PMID- 11486278 TI - Congenic mapping confirms a locus on rat chromosome 10 conferring strong protection against myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis. AB - Myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE) in rats closely mimics the human disease multiple sclerosis (MS). As in MS, genetic predisposition to MOG-EAE is regulated by both MHC and non-MHC genes. Based on disease regulatory influences on MOG-EAE on chromosome 10 in an F2 cross between susceptible DA and resistant ACI rats, we have now isolated this locus in a congenic rat strain to enable further dissection of disease mechanisms. This region is of particular interest, since it is homologous to human 17q for which human whole-genome scans have indicated harbors genes regulating susceptibility to MS. Phenotypic comparison between DA and the congenic DA.ACI-D10Rat2-D10Rat29 strain confirms that the chromosomal segment harbors gene(s) conferring strong protection against MOG-EAE. Furthermore, resistance to EAE in this congenic strain is associated with absence or a low level of inflammation and demyelination in the central nervous system. Levels of anti-MOG antibody isotypes did not differ between parental and congenic rats, thus an action on Th1/Th2 differentiation is unlikely. In conclusion, this is the first example of an EAE-regulating locus isolated in a congenic rat strain with retained phenotype. The mechanism by which gene(s) in the region act is still unclear and will require further studies with this congenic rat strain as a tool. PMID- 11486279 TI - Exon-intron organization of TRGC genes in sheep. AB - A series of genomic clones derived from a sheep library were used to determine the germline configuration and the exon-intron organization of TRGC2, TRGC3, and TRGC4 genes. Based on the outcomes of molecular analysis, we compared and aligned the genomic sequences with the known complete cDNA sequences of sheep and deduced the exon-intron organization of TRGC genes in this ruminant animal, EX1, corresponding to the disulfide-linked constant domain, and EX3, corresponding to the transmembrane and cytoplasmatic domains, are similar in length in all genes. Conversely, the hinge-encoding EX2A, EX2B, and EX2C exons differ in number and length between genes, and EX2A contains the TTKPP motif irrespective of whether it occurs in single or triplicate form. The molecular data also indicate that at least one additional gene is present in sheep. Phylogenetic analysis grouped the ruminant TRGC genes in two clusters that could have emerged from two ancestral forms that underwent a series of duplications giving rise to the new sequences that were selected and then fixed in the ruminant lineages. A correlation between the cluster distribution in the phylogenetic tree of TRGC genes and their expression during fetal development is discussed. PMID- 11486280 TI - A common rhesus macaque MHC class I molecule which binds a cytotoxic T-lymphocyte epitope in Nef of simian immunodeficiency virus. PMID- 11486281 TI - Polymorphisms of the tumor necrosis factor receptor type 1 locus among autoimmune susceptible and resistant inbred rat strains. PMID- 11486282 TI - An Mhc class I gene in the Australian brushtail possum (Trichosurus vulpecula). PMID- 11486283 TI - Fever in returned travelers: review of hospital admissions for a 3-year period. AB - We reviewed 232 consecutive patients admitted to a tertiary-care hospital under the care of an infectious diseases unit for management of febrile illness acquired overseas. A total of 53% presented to hospital within 1 week of return and 96% within 6 months. Malaria was the most common diagnosis (27% of patients), followed by respiratory tract infection (24%), gastroenteritis (14%), dengue fever (8%), and bacterial pneumonia (6%). Pretravel vaccination may have prevented a number of admissions, including influenza (n=11), typhoid fever (n=8) and hepatitis A (n=6). Compared to those who had not traveled to Africa, those who had were 6 times more likely to present with falciparum than nonfalciparum malaria. An itinerary that included Asia was associated with a 13-fold increased risk of dengue, but a lower risk of malaria. Palpable splenomegaly was associated with an 8-fold risk of malaria and hepatomegaly with a 4-fold risk of malaria. As a cause of fever, bacterial pneumonia was > or =5 times more likely in those who were aged >40 years. PMID- 11486284 TI - Empirical therapy for uncomplicated urinary tract infections in an era of increasing antimicrobial resistance: a decision and cost analysis. AB - Infectious Diseases Society of America guidelines state that uncomplicated urinary tract infections (UTIs) should be treated empirically with trimethoprim sulfamethoxazole (TMP-SMZ), unless the community resistance among uropathogens exceeds 10%-20%, in which case a fluoroquinolone (FQ) should be used. However, the data to support this threshold are limited. We performed a cost-minimization and sensitivity analysis to determine what level of TMP-SMZ resistance in a community should trigger FQ use. The mean cost of empirical treatment with TMP SMZ was US$92 when the proportion of resistant Escherichia coli was 0%, $106 when it was 20%, and $120 when it was 40%. The mean cost of empirical FQ treatment was $107 at current levels of FQ resistance. When >22% of E. coli in a community are TMP-SMZ-resistant, empirical FQ therapy becomes less costly than TMP-SMZ therapy. Treatment guidelines for empirical treatment of UTIs may need modification, and the threshold trigger for empirical FQ use should be raised to >20% TMP-SMZ resistance. PMID- 11486285 TI - Infectious complications among 620 consecutive heart transplant patients at Stanford University Medical Center. AB - A total of 1073 infectious episodes (IEs) that occurred in 620 consecutive heart transplantation patients at Stanford Medical Center between 16 December 1980 and 30 June 1996 were reviewed. Infectious complications were a major cause of morbidity and mortality, second only to rejection as the cause of early deaths and the most common cause of late deaths. Of the IEs, 468 (43.6%) were caused by bacteria, 447 (41.7%) by viruses, 109 (10.2%) by fungi, 43 (4.0%) by Pneumocystis carinii, and 6 (0.6%) by protozoa. The largest number of IEs occurred in the lungs (301 [28.1%]). A significant reduction in the incidence of IEs and a delay in presentation after transplantation were observed; these were most likely related to the introduction of new chemoprophylactic regimens during the study period and prevention of significant disease caused by cytomegalovirus. PMID- 11486286 TI - Trends in mortality due to invasive mycotic diseases in the United States, 1980 1997. AB - To determine national trends in mortality due to invasive mycoses, we analyzed National Center for Health Statistics multiple-cause-of-death record tapes for the years 1980 through 1997, with use of their specific codes in the International Classification of Diseases, Ninth Revision (ICD-9 codes 112.4-118 and 136.3). In the United States, of deaths in which an infectious disease was the underlying cause, those due to mycoses increased from the tenth most common in 1980 to the seventh most common in 1997. From 1980 through 1997, the annual number of deaths in which an invasive mycosis was listed on the death certificate (multiple-cause [MC] mortality) increased from 1557 to 6534. In addition, rates of MC mortality for the different mycoses varied markedly according to human immunodeficiency virus (HIV) status but were consistently higher among males, blacks, and persons > or =65 years of age. These data highlight the public health importance of mycotic diseases and emphasize the need for continuing surveillance. PMID- 11486287 TI - What are today's orphaned vaccines? AB - Development costs for new biological agents are increasing, and the time span from laboratory research to introduction of a product on the world market is becoming ever longer. Complex regulatory requirements add barriers and additional costs to early introduction abroad. This results in reluctance by manufacturers to undertake development of a vaccine that will be used for a tropical disease in only the public sector of a poor country. The chances of recovery of huge investment costs before patents expire are not good, unless such a new vaccine can also be sold at high cost in North America and Europe. These are some of the reasons that we still do not have a modern Japanese encephalitis vaccine or products against malaria and dengue fever. Many tropical countries must find a way to develop their own vaccine production facilities. Innovative help for technology transfer will have to be forthcoming, or many new life-saving products will never bridge the gap between research unit and production. PMID- 11486288 TI - Malaria on the World Wide Web. AB - The Internet is enabling scientists and clinicians in areas with endemic malaria to transfer information to scientists and clinicians in other countries. This should allow changes in therapy to follow the rapid changes in the disease that have posed such difficulties in the past. This article reviews Internet resources that focus on malaria. This includes 90 Web sites in 12 sections. Authoritative multinational organizational sites and regional sites, such as those in Africa, Asia (including Thailand and India), and South America (in Venezuela and Brazil), are described. Basic research-oriented databases, such as those that deal with plasmodia genomics, biochemistry, and vaccine development, as well as vector information and geographic satellite information systems, are reviewed. There is a section about malaria research-funding organizations that offer online applications. Useful teaching resources and journals, including those with full online access, are detailed. PMID- 11486289 TI - Rethinking recommendations for use of pneumococcal vaccines in adults. AB - Streptococcus pneumoniae remains a major cause of disease worldwide; the emergence of antibiotic-resistant strains emphasizes the importance of disease prevention by use of vaccines. Recent studies have provided information that is useful for the evaluation of current vaccine recommendations. Recommendations target most people who are at high risk for invasive pneumococcal disease. However, higher risk has also been identified for African Americans and smokers, but these groups are not specifically targeted by current recommendations. The vaccine is effective against invasive disease in immunocompetent people, although studies in immunocompromised subjects have found few subgroups in which the vaccine appears to be effective. Questions with regard to optimal timing and indications for revaccination remain a challenge, because the duration of protection and effectiveness of revaccination remain unknown. New pneumococcal vaccines appear promising but will need to be tested against the performance of the polysaccharide vaccine. Improving delivery of the currently available pneumococcal polysaccharide vaccine to adults who will benefit should be a high priority. PMID- 11486290 TI - Coinfecting deer-associated zoonoses: Lyme disease, babesiosis, and ehrlichiosis. AB - The heightened worldwide recognition of the health burden of tickborne infection derives largely from the increasing incidence of Lyme disease, human babesiosis, and human granulocytic ehrlichiosis, both individually and in concert. Because these infections share the same rodent reservoir and tick vector hosts, they can be cotransmitted to human hosts. Indeed, human coinfections involving various combinations of these pathogens are common, and some tend to be particularly severe. Diagnostic procedures and clinical management of the resulting disease syndrome is rendered complex by the diversity of pathogens involved and by the unusual diversity and duration of symptoms. PMID- 11486291 TI - The malevolent use of microbes and the rule of law: legal challenges presented by bioterrorism. PMID- 11486292 TI - Adherence to highly active antiretroviral therapy predicts virologic outcome at an inner-city human immunodeficiency virus clinic. AB - This study's hypothesis is that human immunodeficiency virus-infected patients in the inner city (predominantly injection drug users and ethnic minorities) do not take highly active antiretroviral therapy (HAART) as prescribed and that nonadherence leads to virologic failure. A prospective, observational, 3-month study of adherence to HAART was undertaken at an inner-city clinic. There were 40 subjects [110 subject-months]; 30 were male, 10 were female, 75% were Hispanic, 23% were African American, 68% were injection drug users, and 68% were receiving triple therapy. At 3 months, adherence, which was determined by use of the Medication Event Monitoring System (Aprex) was significantly associated with virologic success: lower virus loads were associated with a rate of adherence of >80% (P<.05). Although nonadherence predicted virologic failure, virologic success was not always predicted by adherence: 11 (27.5%) of 40 subjects with suboptimal adherence rates (<90%) had complete virologic suppression. PMID- 11486293 TI - Extent of human immunodeficiency virus type 1 drug resistance as a predictor of virological failure after genotype-guided treatment switch. AB - Little is known about factors involved in virological response to treatment changes guided by genotyping in patients whose highly active antiretroviral therapy (HAART) fails. A 12-month observational study was conducted of 45 patients infected with human immunodeficiency virus (HIV)-1, who underwent a new genotype-guided HAART regimen following virological treatment failure. Logistic regression models were used to define factors predictive of virological response to genotype-assisted treatment switches. Virological response was defined as achievement of a level of plasma HIV-1 RNA <1000 copies/mL at the end of the follow-up. Drug-resistance mutations were detected at baseline in 30 patients (66.7%). A sustained virological response to new treatment occurred in 13 (43.3%) of these, as opposed to 11 (73.3%) of the 15 patients harboring drug-susceptible virus at baseline (P=.07). In multivariate logistic regression analysis, the number of drug classes where there was resistance at baseline was the only independent predictor of virological failure (P=.0313). Lack of virological response to genotype-guided treatment changes is primarily due to complex baseline resistance patterns. Benefits of antiretroviral resistance testing may be seriously limited by the lack of subsequent treatment options for heavily pretreated patients. PMID- 11486294 TI - Modifiable dietary habits and their relation to metabolic abnormalities in men and women with human immunodeficiency virus infection and fat redistribution. AB - We assessed the relationship between dietary intake, body composition, and metabolic parameters in 85 consecutive human immunodeficiency virus (HIV) infected patients with fat redistribution. Dietary history and values for fasting glucose, insulin, lipids, and oral glucose tolerance were obtained for 62 men and 23 women with HIV infection and fat redistribution (mean age +/- standard error of the mean [SEM], 43.5+/-0.9 years; mean body mass index [BMI] +/- SEM, 26.3+/ 0.5 kg/m2). A multivariate regression analysis was used to predict insulin area under the curve (AUC) following the oral glucose tolerance test; this included age, sex, BMI, waist-to-hip ratio, kilocalories, duration of protease inhibitor (PI) use, fat redistribution pattern, alcohol intake, dietary fiber intake, and polyunsaturated-to-saturated (P:S) fat ratio. Only age (P=.004), PI use duration (P=.02), and P:S fat ratio (P=.003) were positively associated with insulin AUC. Dietary fiber intake was inversely associated with the insulin AUC (P=.001). In a similar analysis, alcohol consumption was a significant positive predictor of low density lipoprotein cholesterol. Polyunsaturated fats, fiber, and alcohol are strongly associated with insulin resistance and hyperlipidemia in this population and may be important targets for dietary modification. PMID- 11486295 TI - Acute endogenous endophthalmitis due to Bartonella henselae. AB - A 45-year-old man presented with progressively worsening vitreitis of 1 week's duration. Treatment for cat-scratch disease 3 years prior to presentation and persistent vitreitis led to vitrectomy, and analysis of the vitrectomy specimen revealed inflammatory cells and necrotic debris; polymerase-chain-reaction analysis of the vitreous fluid sample, done by use of a novel heminested protocol, demonstrated the presence of Bartonella henselae DNA. Treatment with doxycycline led to improvement in the intraocular inflammation but resulted in a poor visual outcome. PMID- 11486296 TI - Systemic inflammatory response syndrome in adult patients with nosocomial bloodstream infection due to Staphylococcus aureus. AB - To determine the impact of methicillin resistance on clinical course and outcome, we evaluated nosocomial bloodstream infections (BSIs) due to Staphylococcus aureus that were diagnosed in 82 adult patients at the Medical College of Virginia Hospitals from December 1995 through May 1997. Patients with BSI due to methicillin-resistant S. aureus were compared with patients with BSI due to methicillin-susceptible S. aureus; the groups did not differ with regard to inflammatory response or outcome. Mortality was predicted by systemic inflammatory response and Acute Physiology and Chronic Health Evaluation II score but did not correlate with bacterial resistance to methicillin. PMID- 11486297 TI - Necrotizing fasciitis due to Streptococcus pneumoniae after intramuscular injection of nonsteroidal anti-inflammatory drugs: report of 2 cases and review. AB - Two cases of pneumococcal necrotizing fasciitis (NF) occurred after intramuscular injections of nonsteroidal anti-inflammatory drugs; another 5 cases reported in the literature fulfilled the criteria for NF involving Streptococcus pneumoniae. Conditions associated with alterations of immune function could be identified in 6 of the 7 cases; 2 patients died despite surgical and antimicrobial treatment. PMID- 11486298 TI - Failure of treatment for chronic Mycobacterium abscessus meningitis despite adequate clarithromycin levels in cerebrospinal fluid. AB - We report a case of posttraumatic meningitis due to Mycobacterium abscessus, treated initially with oral clarithromycin and intravenous amikacin plus intrathecal amikacin. Despite cerebrospinal fluid (CSF) levels of clarithromycin and amikacin in excess of their in vitro minimum inhibitory concentrations for the organism, the CSF cultures remained continuously positive for M. abscessus. To our knowledge, this is the first documented case of M. abscessus meningitis and the first report of measured CSF levels of clarithromycin in a patient with meningitis, showing that even therapeutic CSF levels of clarithromycin and amikacin might not be successful in eradicating M. abscessus meningitis. PMID- 11486299 TI - Polyclonal proliferation of large granular lymphocytes during cytomegalovirus primary infection in a human immunodeficiency virus-infected patient receiving antiretroviral therapy. AB - We report the first case of a patient infected with HIV in whom polyclonal CD8+/CD57- T lymphocyte large granular lymphocyte (LGL) proliferation was observed in association with cytomegalovirus primary infection. Because the differential diagnosis of an increased number of LGLs includes both monoclonal LGL leukemia and polyclonal proliferation of LGL, patients in whom LGL proliferation is detected always need close hematological and clinical observation to determine whether therapeutic intervention is necessary. PMID- 11486300 TI - Pseudomembranous tracheobronchitis due to Bacillus cereus. AB - We present a case of a rapidly progressive pseudomembranous tracheobronchitis and pneumonia in a 52-year-old woman with severe aplastic anemia. Bacillus cereus was isolated from bronchoalveolar lavage fluids, blood cultures, and pseudomembrane biopsy specimens; despite intensive antibiotic treatment, the patient's condition deteriorated rapidly. To our knowledge, this is the first report of a B. cereus infection that has caused pseudomembranous tracheobronchitis, possibly because of the production of bacterial toxins. PMID- 11486301 TI - Recognition, prevention, surveillance, and management of surgical site infections: introduction to the problem and symposium overview. PMID- 11486302 TI - Surgical site infection (SSI) rates in the United States, 1992-1998: the National Nosocomial Infections Surveillance System basic SSI risk index. AB - By use of the National Nosocomial Infections Surveillance (NNIS) System's surgical patient surveillance component protocol, the NNIS basic risk index was examined to predict the risk of a surgical site infection (SSI). The NNIS basic SSI risk index is composed of the following criteria: American Society of Anesthesiologists score of 3, 4, or 5; wound class; and duration of surgery. The effect when a laparoscope was used was also determined. Overall, for 34 of the 44 NNIS procedure categories, SSI rates increased significantly (P< .05) with the number of risk factors present. With regard to cholecystectomy and colon surgery, the SSI rate was significantly lower when the procedure was done laparoscopically within each risk index category. With regard to appendectomy and gastric surgery, use of a laparoscope affected SSI rates only when no other risk factors were present. The NNIS basic SSI index is useful for risk adjustment for a wide variety of procedures. For 4 operations, the use of a laparoscope lowered SSI risk, requiring modification of the NNIS basic SSI risk index. PMID- 11486303 TI - Maximizing appropriate antibiotic prophylaxis for surgical patients: an update from LDS Hospital, Salt Lake City. AB - Errors in antimicrobial prophylaxis for surgical patients remain one of the most frequent types of medication errors in hospitals. Failure to administer the first dose of antimicrobial prophylaxis within the 2-h window of time before incision is associated with 2- to 6-fold increases in rates of surgical site infection. Optimal use of antimicrobial prophylaxis includes proper case selection; use of appropriate agents; proper dosing, route of administration, timing, and duration; and intraoperative dosing when appropriate. Effective use of antimicrobial prophylaxis also requires monitoring of and feedback on patterns of use. Programs to improve antimicrobial prophylaxis should be multidisciplinary and should aim to improve use of medications, not simply to change physician practice patterns. The LDS Hospital experience demonstrates the clinical and financial benefits of such a program and also shows the pitfalls of and great difficulties associated with changing systems of care. PMID- 11486304 TI - Clinical presentations of soft-tissue infections and surgical site infections. AB - Skin and soft-tissue infections that usually follow minor traumatic events or surgical procedures are caused by a wide spectrum of bacteria. Less frequently, the infections occur spontaneously, which often is clinically confusing and leads to delays in diagnosis. Most of the infections are self-limited and easily treated with local measures and/or antibiotics. Others are life-threatening, requiring prompt diagnosis and aggressive surgical debridement in addition to the wise choice of antibiotic agents to limit tissue loss and preserve life. Many survivors experience critical tissue losses that may require changes in lifestyle as well as major reconstructive cosmetic surgery. Involvement of antibiotic resistant gram-positive microorganisms in these infections only increases the difficulty of their treatment and may have a significant influence on the ultimate outcome. PMID- 11486305 TI - New developments in diagnosis and treatment of infection in orthopedic implants. AB - Orthopedic implants have revolutionized treatment of bone fractures and noninfectious joint arthritis. Today, the risk for orthopedic device-related infection (ODRI) is <1%-2%. However, the absolute number of patients with infection continuously increases as the number of patients requiring such implants grows. Treatment of ODRIs most frequently includes long-term antimicrobial treatment and removal of the implant. Recent evidence from observational trials and 1 randomized clinical trial indicate that a subset of patients can be successfully treated with retention of the implant. Patients eligible for such a treatment must meet the following criteria: acute infection defined as signs and symptoms lasting <14-28 days, an unambiguous diagnosis based on histopathology and microbiology, a stable implant, and susceptibility of the microorganism to an effective orally available antimicrobial agent. PMID- 11486306 TI - Introduction to therapies in multiple myeloma. PMID- 11486307 TI - Standard-dose therapy for multiple myeloma: The Southwest Oncology Group experience. AB - We summarize the Southwest Oncology Group (SWOG) experience with standard therapy for multiple myeloma by reviewing and updating data from seven consecutive SWOG trials. Some modest progress has been made since the introduction of melphalan and prednisone (MP) for induction therapy, using regimens that involve vincristine and doxorubicin, and which save alkylating agents for possible later high-dose therapy. For maintenance, it appears that prednisone plays a useful role. We demonstrate the use of the data collected in these trials with a proposed new staging system. PMID- 11486308 TI - Some early phase II trials in previously untreated multiple myeloma: The Royal Marsden experience. AB - Multiple myeloma is at a stage today where acute leukemia was in the 1960s and 1970s when the aim was to attain complete remission (CR), which would translate into prolonged overall survival (OS) and a cure. The Royal Marsden group was the first to establish a dose-response effect for melphalan leading to CR in patients with myeloma. Since then, different strategies have been developed to improve the results of high-dose therapy, including alternating conditioning regimens, peripheral blood stem cells (PBSC) as a source of stem cells, purging techniques to decrease graft contamination by the myeloma cell, increased dose intensity by means of tandem transplants, and finally, allogeneic matched or unmatched stem cell transplantation. Components of treatment offered at Royal Marsden since 1980 are upfront high-dose melphalan (HDM) 140 mg/m(2), induction infusional chemotherapy followed by consolidation with HDM with autotransplantation, and maintenance with interferon-alpha2b (IFN) (collectively termed sequential therapy). Each of these components has shown benefits in selected studies and together they have dramatically improved the outlook for patients with myeloma. PMID- 11486309 TI - European Group for Blood and Marrow Transplantation Registry studies in multiple myeloma. AB - The European Group for Blood and Marrow Transplantation (EBMT) Myeloma Registry, established in 1987, contains data on 1,368 allogeneic and more than 8,000 autologous stem cell transplants performed since 1983. Among autologous transplant patients, the median survival after transplantation is 50 months, and the actuarial survival at 10 years is 30%, with a plateau appearing at about 8 years. Factors of importance for a more favorable prognosis are lower age, response to chemotherapy, only one course of primary chemotherapy, stage I or II disease, and low beta(2)-microglobulin at diagnosis. Beneficial procedural factors associated with better outcome are a preparative regimen without total body irradiation (TBI), posttransplant interferon alfa maintenance treatment, and possibly tandem transplantation. In vitro graft purging, using CD34(+) selection, does not have any impact on survival. A case-matched analysis comparing autologous and allogeneic transplantation demonstrated significantly better survival in the former group, with median posttransplant survival times of 36 months and 18 months in the autologous and allogeneic groups, respectively. This result was in turn due to a markedly lower incidence of transplant-related death among the autotransplant patients: 13%, versus 41% for the allogeneic group. However, recent data on allogeneic transplants performed from 1994 to 1998 has demonstrated a decrease in treatment-mortality to 30%, and this has resulted in a prolongation of survival; in this analysis, the results are similar irrespective of the type of graft used, allogeneic bone marrow or blood stem cells. In a small case-matched analysis, transplantation with an identical twin donor was superior to both allogeneic and autologous transplantation with respect to survival and freedom from progression. PMID- 11486310 TI - Randomized trial experience of the Intergroupe Francophone du Myelome. AB - This article summarizes clinical results of the Intergroupe Francophone du Myelome (IFM) trials: high-dose therapy (HDT) supported with autologous stem cells improves survival, melphalan 200 mg/m(2) is the best preparative regimen, unpurged peripheral blood stem cells (PBSC) are the recommended source of stem cells to support HDT, and tandem transplants significantly improve survival. Although these results are encouraging, the current IFM 99 protocol will evaluate innovative strategies with the goal to improve long-term survival. PMID- 11486311 TI - Results of high-dose treatment with autologous stem cell support in patients with multiple myeloma. AB - The introduction of high-dose treatment (HDT) with autologous hematopoietic stem cell support in patients with multiple myeloma (MM), although curative in only a minority of patients, has led to significantly longer event-free (EFS) and overall survival (OS) and likely improved quality of life compared with conventional chemotherapy. However, several issues remain unsettled. We review the current knowledge with respect to transplantation-related controversies, such as intensity of cytoreductive therapy (single v tandem transplants), optimal timing of the procedure, and potential benefit of graft purging and posttransplant consolidation chemotherapy. Special emphasis is placed on the safety and efficacy of transplant in elderly patients and in those with impaired renal function, who are usually excluded from the most effective therapy. PMID- 11486312 TI - Allogeneic hematopoietic cell transplantation for multiple myeloma. AB - Some patients with multiple myeloma (MM) who have undergone allogeneic hematopoietic stem cell transplants remain free of disease 5 to 13 years later-a major accomplishment for a malignancy that had been resistant to all investigational therapies. Although it will require longer follow-up to determine how many are truly cured, results for patients with MM transplanted from identical twins suggest that long-term progression-free survival is possible. While 3- to 5-year survival is similar after allogeneic or autologous stem cell transplant for MM, only allograft recipients appear to enjoy long-term disease free survival, most likely due to an allogeneic graft-versus-myeloma (GVM) effect. The very high transplant-related mortality associated with standard allogeneic stem cell transplantation is currently the major limitation to wider use of this potentially curative treatment. The challenge for clinical investigators will be to reduce the incidence of posttransplant complications. Strategies include the use of nonablative conditioning for allografts, the administration of peripheral blood stem cells (PBSC) rather than bone marrow, and the application of more focused, targeted conditioning therapies such as bone seeking radioisotopes. PMID- 11486313 TI - Thalidomide in the management of multiple myeloma. AB - Thalidomide has recently been shown to have significant activity in refractory multiple myeloma (MM). A follow-up of the original phase II trial, expanded to 169 patients, shows 2-year survival of 60%; patients receiving > or =42 g over 3 months had a higher response rate and superior survival than those receiving lower doses. The addition of thalidomide to dexamethasone and chemotherapy for the management of post-transplant relapses results in higher response rates. The early results of the Total Therapy II trial for newly diagnosed MM patients show an unprecedented complete remission (CR) and near-CR rate of 69% after two melphalan-based transplants (whether or not receiving thalidomide). In addition, available clinical trial information involving at least 20 patients confirms that thalidomide is active in one third of patients in single-agent trials for refractory disease, with response rates increasing to 50% to 60% in combination with dexamethasone and to as high as 80% in combination with dexamethasone and chemotherapy. When applied as primary therapy in smoldering myeloma, one third of patients experienced 50% paraprotein reduction (PPR); in combination with dexamethasone pulsing, 70% to 80% of symptomatic patients responded. Thus, thalidomide is a major new tool in the treatment armamentarium of MM. The virtual lack of myelosuppression makes it an ideal agent for combination with cytotoxic chemotherapy. Newer, more potent, and less toxic derivatives of thalidomide are being evaluated. PMID- 11486314 TI - Immunotherapy of multiple myeloma. AB - The failure of chemotherapy to cure a significant proportion of multiple myeloma (MM) patients and increasing knowledge of tumor immunology and MM biology have generated considerable interest in immunotherapy for this lethal disease. Immunotherapy for MM can be divided into three broad categories: passive antibody mediated immunotherapy, active specific immunization (vaccination), and adoptive T-cell immunotherapy. Early clinical trials using anti-CD20 monoclonal antibodies (mAbs) have met with limited success so far but have also suggested that selected patient subgroups may benefit from this treatment. The availability of a truly tumor-specific antigen such as immunoglobulin idiotype, the recent demonstration that MM cells process and present idiotype to T lymphocytes, and formal evidence of an antitumor effect of idiotypic vaccination in follicular lymphoma provide the framework for applying idiotypic vaccination in MM. The ability to generate ex vivo functional dendritic cells has made it possible to fuse them with patients' MM cells, thus producing a different type of customized vaccine. Dendritic cells are also a pivotal reagent to generate ex vivo MM-specific cytotoxic T lymphocytes (CTLs) to be reinfused into the patient for adoptive immunotherapy. This review summarizes achievements in MM immunotherapy based on data reported since 1998. PMID- 11486315 TI - Gene therapy approaches for multiple myeloma. AB - Several different myeloma gene therapy approaches are currently being explored, seeking to impact on the disease process in diverse ways. Therapeutic benefit may result from destroying the myeloma cells directly, provoking an antimyeloma cell immune response, interfering with the paracrine growth signaling pathways between osteoclasts and myeloma cells, or genetically manipulating hematopoietic progenitors or mature T cells in a stem cell transplantation setting. Encouraging progress in each of these areas is being fueled by the development of improved viral and nonviral gene transfer vectors. PMID- 11486316 TI - Myeloma bone disease. AB - Bone destruction is a hallmark of myeloma, with 70% to 80% of patients manifesting bone involvement. Destruction is mediated through normal osteoclasts (OCLs), which respond to local osteoclast-activating factors (OAFs) produced by myeloma cells or by other cells in the local microenvironment. OAFs implicated in myeloma bone disease include tumor necrosis factor-beta (TNFbeta), RANK ligand (RANKL), interleukin-1 (IL-1), parathyroid hormone-related protein (PTHrP), hepatocyte growth factor (HGH), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNFalpha), and macrophage inflammatory protein-1-alpha (MIP-1alpha). To date, the leading candidates for OAFs are MIP-1alpha and RANKL. Adhesive interactions between marrow stromal cells and myeloma cells induce marrow stromal cells to secrete IL-6, a potent myeloma growth/survival factor that may contribute to the bone disease. Evaluation of myeloma bone disease includes plain radiographs, and newer methods, such as magnetic resonance imaging (MRI), positron emission tomography (PET) scans, technetium-99m-sestamibi (Mibi) scanning, and dual-energy x-ray absorptiometry (DEXA) scanning, may provide more complete information. In addition, biochemical markers of bone resorption are being evaluated, although the limited availability of these assays and lack of extensive testing in patients make their routine use premature. Treatment of myeloma bone disease includes radiation therapy, vertebroplasty, surgery, and bisphosphonates. New developments on the pathogenesis and treatment of myeloma bone disease present great opportunities to combat bone disease. PMID- 11486317 TI - Targeted therapy for multiple myeloma. AB - Multiple myeloma (MM) remains incurable with conventional treatment approaches, and novel biologically based therapies are therefore urgently needed. Targeted therapies are either under development or already undergoing clinical evaluation predicated upon: identifying genetic abnormalities in myeloma cells to enhance chemoradiosensitivity; interrupting growth or triggering apoptotic signaling cascades in tumor cells; treating both the tumor cell and its microenvironment; enhancing allogeneic and autologous antimyeloma immunity; and characterizing new myeloma antigens for serotherapy. These therapies, alone or in combination with conventional treatments, offer great promise to improve the outcome for patients with MM. PMID- 11486318 TI - Developmental therapeutics for multiple myeloma--a road map for the future. PMID- 11486319 TI - Outcome of pregnancies in women with uterine leiomyomas identified by sonography in the first trimester. AB - PURPOSE: We assessed the outcome of pregnancies in women with uterine leiomyomas (fibroids) documented by sonography in the first trimester of pregnancy. METHODS: We collected cases of women who had undergone first-trimester sonography and had uterine fibroids and singleton pregnancies with documented fetal heartbeats. We compared pregnancy loss rates and modes of delivery in these cases to a maternal age-matched and gestational-age-matched control group of women who had normal uteruses and first-trimester pregnancies with documented fetal heartbeats. Sonograms in patients with fibroids were reviewed to determine the number of fibroids, their sizes, and their locations. Within the group of patients with fibroids, the pregnancy loss rate was also compared based on the number of fibroids and fibroid size and location. RESULTS: Our study population consisted of 143 women with leiomyomas, and our control group comprised 715 patients with a normal uterus. Among patients with fibroids, 14.7% of pregnancies resulted from assisted conception; in the control group, 6.4% of pregnancies resulted from assisted conception. The rate of spontaneous pregnancy loss in women with fibroids was almost twice the rate in women with normal uteruses (14.0% versus 7.6%; p < 0.05), and the loss rate was higher in women with multiple fibroids than in women with a single leiomyoma (23.6% versus 8.0%, p < 0.05). The loss rate was not significantly associated with fibroid size or location. The rate of cesarean-section delivery was higher in patients with fibroids than in patients with normal uteruses (38% versus 28%, p < 0.05). CONCLUSIONS: Uterine fibroids are associated with an elevated risk of spontaneous pregnancy loss. The loss rate is higher in patients with multiple fibroids than with a single fibroid. The cesarean-section rate is also higher in patients with fibroids than in patients with a normal uterus. PMID- 11486320 TI - Impact of intraoperative sonography on resection and cryoablation of liver tumors. AB - PURPOSE: We retrospectively analyzed the impact of intraoperative sonography (IOUS) on the management of patients referred for resection of liver tumors. METHODS: Forty patients underwent IOUS with a 7-MHz curved-array sector transducer; in selected cases, a 5-MHz linear-array transducer attached to a color Doppler unit was also used. The number, size, and location of tumors on IOUS, including tumor proximity to or invasion of major vessels or invasion of the diaphragm, were compared to findings on preoperative imaging studies. The effect of these findings on surgical management was assessed. Unresectable lesions were treated by cryoablation under ultrasound guidance. RESULTS: IOUS detected preoperatively unsuspected lesions in 7 patients (18%). Metastases suspected on CT arterial portography were ruled out in 2 patients (5%), and indeterminate lesions were diagnosed as cysts by IOUS in 2 other patients (5%). Vascular proximity or vascular or diaphragmatic invasion detected by IOUS rendered lesions unresectable in 4 patients (10%). Cryoablation under IOUS guidance and monitoring was attempted in 11 patients (28%) and performed successfully in 10. CONCLUSIONS: IOUS changed the management in 38% of patients and guided cryoablation in 28% of patients. IOUS performed by an experienced sonologist is invaluable for the accurate assessment of liver tumor resectability; the detection of additional, preoperatively unknown lesions; and the guidance of cryoablation of unresectable tumors. PMID- 11486321 TI - Doppler sonography of the penile cavernosal artery: comparison of intraurethral instillation and intracorporeal injection of prostaglandin E1. AB - PURPOSE: We compared the Doppler sonographic findings in the penile cavernosal artery (arteria profunda penis) after intraurethral instillation and intracorporeal injection of prostaglandin E1 (PGE1) to evaluate the hemodynamic changes during drug-induced erection. METHODS: Twenty healthy male volunteers were enrolled in the study. Ten subjects (intraurethral group) were examined with Doppler sonography of the penile cavernosal artery after intraurethral administration of 1 mg of PGE1. The remaining 10 subjects (intracorporeal group) underwent Doppler sonography of the cavernosal artery after intracorporeal injection of 5 microg of PGE1. The peak systolic velocity, minimal diastolic velocity, and resistance index were determined at 5-minute intervals for 30 minutes following administration of PGE1 in both groups. The results were compared between the 2 groups. RESULTS: The peak systolic velocity in the intraurethral group increased progressively from a mean of 31.1 cm/second at 5 minutes to 65.6 cm/second at 30 minutes after intraurethral administration of PGE1. In the intracorporeal group, the mean peak systolic velocity ranged from 44.1 to 83.2 cm/second, reached a maximum at 10 minutes, and then decreased continuously through 30 minutes after intracorporeal injection of PGE1. The mean peak systolic velocities were significantly higher in the intracorporeal group at 10 and 15 minutes (p < or = 0.05); the mean minimal diastolic velocities were significantly lower in the intracorporeal group at 15, 20, and 25 minutes (p < or = 0.05); and the mean resistance indices were significantly higher in the intracorporeal group at all time points except 5 minutes (p < or = 0.05). CONCLUSIONS: The intracorporeal injection of PGE1 produced a greater vasoactive response in the cavernosal artery than did intraurethral instillation. PMID- 11486322 TI - Sonographic features of xanthogranulomatous pyelonephritis. AB - PURPOSE: The purpose of this study was to describe the various sonographic features of xanthogranulomatous pyelonephritis (XGP). METHODS: We retrospectively reviewed the CT, sonographic, and medical records of patients diagnosed with XGP from January 1981 to December 1998. Twenty-seven patients for whom XGP was histopathologically confirmed were included in the study. There were 12 men and 15 women, with an age range of 21-86 years (mean, 57 years). All patients had undergone sonography of the kidneys. The renal size, shape, and outline were recorded. The presence of perinephric fluid accumulation, of obstructive uropathy, or of internal echoes in the dilated collecting system and the echotexture of the renal parenchyma were documented. RESULTS: We categorized the XGP into 4 groups on the basis of the sonographic features: (1) diffuse hydronephrotic, 12 patients (44%); (2) diffuse parenchymal, 9 patients (33%); (3) diffuse contracted, 4 patients (15%); and (4) segmental or focal, 2 patients (7%). A localized perinephric fluid collection was present in 4 patients (15%). The preoperative sonographic diagnoses were pyonephrosis (n = 14, 52%), renal pelvic tumor with possible associated infection (n = 5, 19%), renal parenchymal mass (n = 2, 7%), hydronephrosis (n = 2, 7%), and chronic pyelonephritis with renal atrophy (n = 4, 15%). XGP was considered a possible diagnosis in only 11 patients (41%). CONCLUSIONS: XGP has no specific sonographic features but is suggested by parenchymal thinning and hydronephrosis, sonographic signs of chronic obstructive uropathy caused by stones; echoes in the dilated collecting system; and a perinephric fluid collection. CT, needle biopsy, or both are recommended to further evaluate and confirm sonographically suspected XGP. PMID- 11486323 TI - Sonographic evaluation of gallbladder-wall thickening in hemorrhagic fever with renal syndrome: prediction of disease severity. AB - PURPOSE: Gallbladder-wall thickening (GBWT) frequently occurs in patients with hemorrhagic fever with renal syndrome (HFRS), an acute infectious disease caused by hantaviruses. HFRS is manifested by fever, hemorrhage, renal failure, and in many cases gastrointestinal symptoms, such as abdominal pain and tenderness. The clinical significance of GBWT in HFRS has not been reported. The purpose of this study was to investigate the incidence of GBWT and the relationship between GBWT and the severity of HFRS. METHODS: We retrospectively reviewed the medical records and sonograms of 68 patients with HFRS (47 males and 21 females, with an age range of 10-76 years) who underwent abdominal sonography in the acute stage of the disease. We measured the gallbladder-wall thickness on the sonograms and reviewed other sonographic and radiographic findings. Clinical factors that reflect the severity of HFRS were compared between the patients with GBWT (defined as thickness of 4 mm or more) and those without GBWT. RESULTS: Of the 68 patients, 29 (43%) had GBWT, which was even and diffuse in all cases. The patients with GBWT had a significantly lower mean platelet count and serum albumin level and significantly higher serum aspartate aminotransferase and serum lactate dehydrogenase levels than did the patients without GBWT. In addition, the incidence of renal failure requiring hemodialysis and the incidences of ascites and pleural effusion were higher in the patients with GBWT than in those without GBWT. Five patients died of HFRS; all 5 had GBWT (p = 0.011 for comparison with patients without GBWT). CONCLUSIONS: Our results suggest that the sonographic measurement of gallbladder-wall thickness during the acute stage of HFRS is useful for determining the severity of HFRS. PMID- 11486324 TI - Power Doppler sonography of hepatocellular carcinomas with portal-vein blood supply. AB - We report the power Doppler imaging (PDI) findings in 2 cases of hepatocellular carcinoma (HCC) with a portal-vein blood supply. Gray-scale sonography in both cases showed a well-circumscribed nodule, hypoechoic in case 1 and hyperechoic in case 2. PDI revealed an afferent tumor vessel with constant flow in both nodules, and CT during arterial portography demonstrated a portal-vein supply to both nodules. The nodules were diagnosed by percutaneous core biopsies as highly differentiated HCC. We also examined with PDI another 64 patients with 76 HCC nodules, and none of the nodules showed a constant-flow afferent tumor vessel. The presence of a constant-flow afferent tumor vessel indicates a supplying portal vein; but this is not diagnostic of HCC, and biopsy remains necessary to establish a final diagnosis. PMID- 11486325 TI - Partially reversed intrasplenic venous blood flow detected by color Doppler sonography in two patients with hematologic diseases and splenomegaly. AB - We report on 2 patients with hematologic diseases (1 follicular lymphoma and 1 myeloproliferative syndrome) and splenomegaly who had partial intrasplenic portosystemic shunting demonstrated by color Doppler sonography. Intrasplenic venous blood flow was in the normal direction at the hilum of the spleen but in a reversed direction at the periphery of the spleen. This type of reversed intrasplenic flow pattern results in portosystemic shunting and might be detected more frequently when careful color Doppler mapping of the entire splenic parenchyma is performed in patients with portal hypertension. The clinical significance of this phenomenon, however, is still unclear. PMID- 11486326 TI - Sonographic findings in intussusception caused by a lipoma in the muscular layer of the colon. AB - Lipomas of the gastrointestinal tract are uncommon tumors. Almost all gastrointestinal lipomas are submucosal or subserosal, and most are asymptomatic, although they may cause abdominal pain, bowel obstruction, and gastrointestinal bleeding. The diagnosis of gastrointestinal lipoma is usually not made before surgery. We present a case of colonic intussusception caused by a lipoma that was located in the muscular layer of the colon and was diagnosed preoperatively by sonography. The descending colon appeared edematous and thick. There was layering within the lumen of the descending colon, mimicking the target sign. At the distal end of the intussusception, there was a 4.7-cm, hyperechoic, rounded lesion with a smooth margin. Surgery revealed a polypoid mass originating from the splenic flexure and causing intussusception of the colon, and pathologic analysis confirmed the diagnosis of lipoma of the muscular layer of the colon. PMID- 11486327 TI - Intraoral sonographic evaluation of parapharyngeal space tumors. AB - We performed transcutaneous (external) sonography and then intraoral sonography to evaluate 3 parapharyngeal space tumors (1 vagal paraganglioma, 1 pleomorphic adenoma, and 1 carcinoma arising in a pleomorphic adenoma). All 3 patients had medial displacement of a tonsil. In all cases, we could measure the size and see the borders of the tumors and their relation to other structures on intraoral sonography. Intraoral sonography is useful for differentiating between unilaterally enlarged and medially displaced tonsils. PMID- 11486328 TI - Rectus sheath hematoma in pregnancy. AB - Although spontaneous hemorrhage into the sheath of the rectus abdominis muscle is uncommon in pregnancy, rectus sheath hematomas (RSHs) should be considered in patients who present with an acute onset of abdominal pain in the latter half of pregnancy or the immediate postpartum period. Both sonography and CT are useful in diagnosing RSHs. We report a case of pregnancy-associated RSH initially suspected of being a degenerating leiomyoma or torsed ovary. Sonography showed a large mass of mixed echogenicity with no internal vascularity. CT confirmed that the lesion was suprafacial. PMID- 11486329 TI - Morphological and immunophenotypic features of chronic lymphocytic leukemia. AB - In this review, we summarize the morphological features and immunophenotypic profile of chronic lymphocytic leukemia (CLL) cells, discuss the value of these investigations as front line diagnostic tests, and emphasize their correlation with the clinical features, disease progression, molecular genetics and pathogenesis of CLL. In CLL, the morphology of the circulating cells is characteristic and typical in the majority of cases. However, 15% of patients, either at diagnosis or during the course of the disease, show atypical morphology reflected by either (1) an increased (> 10%) number of circulating prolymphocytes, designated CLL/PL, or (2) an increased (> 15%) number of circulating lymphoplasmacytic and cleaved cells, designated 'atypical' CLL. There is strong evidence of a close association between atypical morphology (CLL/PL) and atypical (CLL) and clinical features, e.g. disease progression, advanced stage and survival, molecular genetics, particularly trisomy 12, but also the rare cases with t(11;14) or t(14;19), p53 abnormalities, unmutated immunoglobulin (Ig) VH genes and origin of the cell (naive, pregerminal center cell). CLL cells have a distinct immunological repertoire different from that of other lymphoproliferative disorders. The typical CLL phenotype is CD5+, CD23+, FMC7-, weak expression of surface Ig (sIg) and weak or absent expression of membrane CD22 and CD79b. The latter marker identifies an extracellular epitope of the B cell receptor (BCR) beta chain and its weak or absent expression in CLL may derive from the expression of a truncated form. This, together with the low expression of CD22, might explain the abnormal signal transduction of CLL cells similar to that of anergic B lymphocytes. Because no single marker is specific for CLL, a composite phenotype considering this set of 5 or 6 markers compounded into a scoring system helps to distinguish CLL from the other B-cell malignancies. Immunophenotypic analysis has also been shown to be useful for minimal residual disease detection and adds valuable prognostic information because the expression of certain markers, such as FMC7 or CD38, seems to be associated with a poor outcome. In addition, CLL cells express a variety of Bcl-2 family proteins with a profile that favors inhibition of apoptosis which, together with the interaction with microenvironmental (e.g. stromal) cells and the release of cytokines, explains the long life span and subsequent accumulation of CLL cells in various organs. Despite controversies relating to the expression of adhesion molecules (selectins and integrins) in CLL cells, it appears that some of these molecules do play a role in the pathogenesis, biology and clinical patterns of the disease. In conclusion, morphology and immunophenotype are the two essential investigations, which must be carried out in all cases of CLL. Both provide relevant information in terms of diagnosis, course of the disease, prognosis and pathogenesis. PMID- 11486330 TI - Genetic features of B-cell chronic lymphocytic leukemia. AB - The genetic features of B-cell chronic lymphocytic leukemia (CLL) are currently being reassessed by molecular cytogenetic techniques such as fluorescence in situ hybridization (FISH). Conventional cytogenetic studies by chromosome banding are difficult in CLL mainly because of the low in vitro mitotic activity of the tumor cells, which leads to poor quantity and quality of metaphase spreads. Molecular genetic analyses are limited because candidate genes are known for only a few chromosomal aberrations that are observed in CLL. FISH was found to be a powerful tool for the genetic analysis of CLL as it overcomes both the low mitotic activity of the CLL cells and the lack of suitable candidate genes for analysis. Using FISH, the detection of chromosomal aberrations can be performed at the single cell level in both dividing and non-dividing cells, thus circumventing the need of metaphase preparations from tumor cells. Probes for the detection of trisomies, deletions and translocation breakpoints can be applied to the regions of interest with the growing number of clones available from genome-wide libraries. Using the interphase cytogenetic FISH approach with a disease specific set of probes, chromosome aberrations can be found in more than 80% of CLL cases. The most frequently observed abnormalities are losses of chromosomal material, with deletions in band 13q14 being the most common, followed by deletions in 11q22-q23, deletions in 17p13 and deletions in 6q21. The most common gains of chromosomal material are trisomies 12q, 8q and 3q. Translocation breakpoints, in particular involving the immunoglobulin heavy chain locus at 14q32, which are frequently observed in other types of non-Hodgkin's lymphoma, are rare events in CLL. Genes affected by common chromosome aberrations in CLL appear to be p53 in cases with 17p deletion and ataxia telangiectasia mutated (ATM), which is mutated in a subset of cases with 11q22-q23 aberrations. However, for the other frequently affected genomic regions, the search for candidate genes is ongoing. In parallel, the accurate evaluation of the incidence of chromosome aberrations in CLL by FISH allows the correlation of genetic abnormalities with clinical disease manifestations and outcome. In particular, 17p abnormalities and deletions in 11q22-q23 have already been shown to be among the most important independent prognostic factors identifying subgroups of patients with rapid disease progression and short survival. In addition, deletion 17p has been associated with resistance to treatment with purine analogs. Therefore, genetic abnormalities may allow a risk assessment for individual patients at the time of diagnosis, thus giving the opportunity for a risk-adapted management. PMID- 11486331 TI - Accessory cells, cytokine loops and cell-to-cell interactions in chronic lymphocytic leukemia. AB - In addition to the extensive work that has been conducted in order to understand better the biological features of the leukemic population in B-cell chronic lymphocytic leukemia (CLL), over the years considerable interest has been directed towards other related studies that may have important implications for the accumulation of the leukemic clone and for the immunoparesis typical of this disease. In the present review article, we discuss some of these areas of investigation and, in particular, we focus on: (1) the multiple abnormalities recorded within the T and cytotoxic compartment of patients with CLL; (2) cytokine loops occurring in this disease, with particular emphasis on the cytokines that appear to play a more critical role; and (3) the cell-to-cell cross talk that may be actively operational in CLL. These findings will be discussed in relation with the possible implications that each of them have in the expansion and clinical behavior of a disease that is increasingly proving its heterogeneity. PMID- 11486332 TI - Photodynamically induced changes of acetylcholinesterase activity from human erythrocytes. AB - The light and photodynamic actions on acetylcholinesterase activity from human erythrocytes were studied. After light irradiation (670 nm, semiconducting low power laser) the maximum reaction rate increased from 13.3 to 14.8 mumol per ml packed cells per min for an energy dose 9 J, and above that dose it decreased (10.8 mumol per ml packed cells per min for an energy dose of 15 J). The Michaelis-Menten constant changed in the opposite direction. After irradiation of erythrocyte suspension in the presence of zinc phthalocyanine the reaction rate increased, reaching the maximum for energy dose of 0.75 J (16.85 in comparison to the control value of 14.7 mumol per ml packed cells per min. Similarly, the Michaelis-Menten constant decreased reaching a minimum for an energy dose of 0.75 J (0.04 mM compared to 0.07 mM for control). Incubation of erythrocytes with the dye in the dark increased the reaction rate from 13.3 to 14.7 mumol per ml packed cells per min. Neither the incubation in the dark nor irradiation with laser light caused changes of enzyme activity in the presence of chloroaluminium or metal-free phthalocyanine. PMID- 11486333 TI - Fractionated illumination for oesophageal ALA-PDT: effect on blood flow and PpIX formation. AB - The effect of fractionating the 633 nm illumination of 5-aminolaevulinic (ALA) based photodynamic therapy (PDT) of the normal rat oesophagus was studied. Fractionation of the illumination could enhance the PDT effect in two ways: (a) delay of the vascular shutdown or relaxation of the vasoconstriction induced by ALA-PDT and (b) use of newly formed protoporphyrin IX (PpIX), produced during the dark interval. Forty rats were randomly allocated to two groups of 20 animals each. To study vascular effects, in group 1 illumination with 633 nm (100 mW/cm) was performed at 3 h after oral ALA administration (200 mg/kg) either continuously with 20 J/cm diffuser length (n = 5) or fractionated 2 x 10 J/cm with a 150 s interval (n = 5), five animals served as controls. Blood flow was measured with a laser Doppler flowmeter. To study the effect of renewed PpIX forming, animals in group 2 were illuminated continuously at 3 h after ALA with 20 J/cm (n = 5) or 40 J/cm (n = 5) or fractionated 2 x 20 J/cm with a 3 h interval (n = 5), five animals served as controls. In all animals the in vivo fluence rate and PpIX fluorescence were measured during illuminations and animals were killed at 48 h after PDT. ALA-PDT did not cause any significant vasoconstriction. Fluorescence measurements and dosimetric results in group 1 did not differ between animals illuminated continuously or fractionated with a 150 s interval. In group 2, during a 3 h dark interval, PpIX fluorescence increased and was bleached during the second illumination. The tissue optical properties changed during the 3 h dark interval, resulting in a lower in vivo fluence rate (p < or = 0.001). Fractionation did not result in more oesophageal damage. It was concluded that a 150 s interval during illumination in ALA-PDT does not increase oesophageal blood flow. During an interval of 3 h new PpIX is formed. In the present study, fractionated illumination using short or long time intervals did not result in more damage. Thus, this study shows no evidence for improved PDT effect with fractionated light delivery. PMID- 11486334 TI - Analysis of near-infrared Raman spectroscopy as a new technique for a transcutaneous non-invasive diagnosis of blood components. AB - Near-infrared Raman spectroscopy can be a new technique for physical evaluations, allowing the measurement of lactic acid concentrations, in blood or muscles, during the physical activity in a transcutaneous non-invasive way. Lactic acid accumulation in the human body is one of the factors that leads to fatigue and therefore it should be continually monitored during physical training. Our proposal is to use Raman spectroscopy to monitor the lactic acid present in an athlete without interrupting his exercise for sample collection. The experimental set-up for Raman spectroscopy comprised a near infrared laser at 830 nm, a Kaiser f/1.8 spectrometer and a liquid nitrogen cooled CCD detector. The radiation from the exciting laser is blocked in the collecting system by Kaiser holographic filters. A personal computer controls the entire system, saving and processing the Raman spectra. Experiments were undertaken to verify the presence of lactic acid in the Raman spectra of solutions of lactic acid in human serum and in blood from a Wistar rat. After these two experiments, another was developed in vivo in a Wistar rat, injecting intraperitoneally 1 ml of a 0.12 mol/l lactic acid aqueous solution. An optical fibre catheter touching the skin of the rat groin, over the ileac vein collected the Raman signal. The presence of lactic acid was detected inside a live organism, in a transcutaneous non-invasive way. The minimum lactic acid concentration that the equipment can detect was also studied. An experiment was undertaken for that purpose, in which the laser illuminated directly a quartz cuvette containing solutions with decreasing lactic acid concentrations up to values near to the physiological level in the human body. The results indicated that the technique can be suitable for the physical evaluation of athletes. PMID- 11486335 TI - Interstitial laser photocoagulation and interstitial photodynamic therapy of normal lung parenchyma in the pig. AB - Interstitial laser photocoagulation (ILP) and interstitial photodynamic therapy (PDT) involve delivery of light to lesions in solid organs using thin fibres passed through needles inserted percutaneously under image guidance. In ILP, the laser energy heats the tissue, whereas in PDT it activates a previously administered photosensitising agent. This study looks at their potential for treating localised, small, peripheral lung cancers in patients unsuitable for surgery. Experiments were undertaken on nine normal pigs, up to four fibres being inserted into the lung parenchyma percutaneously under X-ray guidance (ILP: 2-3 W, 1000 q/fibre, from 805 nm diode laser, PDT, 100-200 J/fibre from 652 nm diode laser at 50-100 W, 3 days after 0.15 mg/kg mTHPC). Animals were killed from 3 days to 3 months later and the treated areas examined macroscopically and microscopically. Both techniques were well tolerated, producing well-defined, localised lesions, typically 3.5 x 2 x 2 cm using four fibres. Histology showed thermal coagulative necrosis after ILP and haemorrhagic necrosis after PDT. Early small haematomas and late cavitation were sometimes seen after ILP, but not after PDT. PDT lesions healed with preservation of larger arteries and bronchi in the treated area. A few small pneumothoraces were seen which resolved spontaneously, probably related to the chest wall puncture. It was concluded that ILP and PDT lesions of a size large enough to cover a small tumour can be made safely in the lung parenchyma, although healing was better after PDT. Pilot clinical studies with both techniques are now justified on carefully selected patients. PMID- 11486336 TI - Visual observation of infrared laser speckle patterns at half their fundamental wavelength. AB - In the course of work aimed to examine second harmonic generation in biological macromolecules, it was discovered that 1064 nm pulsed nanosecond infrared laser radiation, after scattering on the optically rough sample surface, was perceived as double the fundamental frequency green speckle pattern by the unaided eye. The threshold of this arresting phenomenon was found to be around 60 mJ/cm2 on hyaluronic acid potassium salt in dark ambient conditions. This effect is of a second order non-linear nature. After experiments with nearsighted, farsighted and normal vision observers in combination with second harmonic generation experiments on various lamb eye tissues, the effect was mainly attributed to second harmonic generation in the cornea arising from phase interference of the incident infrared radiation. The current work could possibly have implications in the fields of speckle interferometry and biomedicine. PMID- 11486337 TI - Er:YAG laser ablation of cerebellar and cerebral tissue. AB - With the availability of suitable fibres, the Er:YAG laser has become an indispensable tool for invasive neurosurgical applications as a source of precise ablation. The aim of this study was to investigate the ablative effects of the Er:YAG laser on brain tissue. The response of neuronal tissue to 2.94 microns Er:YAG laser irradiation was investigated on excised rat brain specimens. Ablation craters were created in cerebral and cerebellar tissues using 0.3, 0.5 and 1.0 J single pulses of 150 microseconds duration. The corresponding average irradiances were 37.7 J/cm2, 62.9 J/cm2 and 125.8 J/cm2, respectively. Craters were checked qualitatively, crater dimensions were measured and compared, and volume of ablated tissue was estimated. Laser-induced crater dimensions were found to be significantly different at different energy levels applied. Moreover, dimensions of craters on cerebral and cerebellar tissues were significantly different in terms of dimensions. We observed that with the Er:YAG laser ablation craters were created with practically no thermal damage to adjacent tissues. The differences observed in the response of cerebral and cerebellar cortical tissues were dependent on the anatomical and chemical differences. PMID- 11486338 TI - Dose-dependency of low-energy HeNe laser effect in regeneration of skeletal muscle in mice. AB - We evaluated the effect on mice skeletal muscle regeneration of different doses (2.6, 8.4, and 25 J/cm2) of HeNe laser (lambda 632.8 nm; power, 2.6 mW; spot size, 0.007 cm2) applied directly to intact skin of injured muscle. Muscle injury was induced in both right and left Tibialis anterior (TA) muscles by ACL myotoxin (5 mg/kg). Right TA muscles were irradiated daily for 5 days while contralateral muscles received a sham treatment. Only the 2.6 J/cm2 dose resulted in changes such as increased mitochondrial density and muscle fibre in the TA muscles as compared to sham groups (3280 +/- 704 microns 2 versus 2110 +/- 657 microns 2, p = 0.02). We concluded that the HeNe effect on mouse muscle regeneration is dose specific: only 2.6 J/cm2 increased muscle fibre area and mitochondrial density. PMID- 11486339 TI - Improvements in colour fundus imaging using scanning laser ophthalmoscopy. AB - The objective of this study was to examine the properties of two laser lines 514 nm and 532 nm when used to image the retina with a scanning laser ophthalmoscope (SLO), and to compare the images taken with a simultaneous multiple wavelength SLO, to those taken with a fundus camera. From this we concluded that the 514 nm line is the preferred line for visualising the nerve fibre layer whereas the 532 nm line is preferred for visualising retinal vessels. Based on these results the 532 nm laser light source was selected as the green line for imaging with the simultaneous colour SLO. Cases are presented where the colour SLO images contain more information than traditional digitised fundus photographs. PMID- 11486340 TI - Neurotrophic factors: important regulators of nociceptive function. AB - Neurotrophic factors have an established developmental role in regulating the survival and specification of sensory neurons. However, these factors continue to exert an important influence on sensory neurons throughout the postnatal period and into adult life. In adulthood, approximately one-half of nociceptors are dependent on nerve growth factor (NGF) for trophic support, whereas the other half are sensitive to glial cell line-derived neurotrophic factor (GDNF). It is now known that many chronic pain states are maintained by widespread changes in the anatomy, neurochemistry, and function of the sensory nervous system both at the level of the primary sensory neuron and the dorsal horn of the spinal cord. Trophic factors appear to orchestrate many of these dynamic changes. This review highlights some of the key roles played by these molecules and in particular the role of NGF in the peripheral sensitization of nociceptors and brain-derived neurotrophic factor (BDNF) as a central pain modulator. PMID- 11486341 TI - The challenges and rewards of integrating diverse neuroscience information. AB - The design of database models and schemas for storing, cross-referencing, and retrieving neuroscience information faces issues that are similar but more complex than most of the other biomedical disciplines, such as genomics and proteonomics. Specifically, the visualization and manipulation of very large and diverse image data, such as digital brain atlases and functional magnetic resonance images, play a unique role in neuroscience while much of the associated information is textually recorded. Nongraphical information can include the annotation of large brain structures ranging from anatomical regions to intracellular structures, the description of cellular functional properties, and their various interrelationships, such as fiber connections. It is necessary that the heterogeneous and distributed types of data be cross-referenced to each other so that this diverse information can be efficiently retrieved, shared, and exchanged among the different neuroscientific disciplines. Continued advances in computers and Internet technologies appear to indicate that increasingly large data sets will be maintained on local or regional file servers and that informational interoperability will be achieved using a networked information system infrastructure. The authors and others have proposed and implemented models of semantically organized information systems that utilize centrally stored and highly structured archival information to index, cross-reference, and retrieve diverse, Web-based data sets. PMID- 11486342 TI - Transplantation and gene therapy: combined approaches for repair of spinal cord injury. AB - Motor and sensory functions are lost after spinal cord injury because neurons die or atrophy and axons fail to regenerate. Until fairly recently, it was believed that damaged neurons could not be replaced and injured axons could not regenerate, and, therefore, functions dependent on injured neurons could not be recovered. We now know that damaged neurons can be rescued by providing therapeutic factors or replaced by grafting. In addition, the adult CNS contains a population of precursor cells with a potential to generate new neural cells, whose numbers and composition can be modified by extrinsic factors. The pioneering studies of Aguayo demonstrated that CNS axons could regenerate in the right environment. Subsequent studies have revealed the identity of some of the inhibitory molecules in myelin and scar tissue, and we now have a better understanding of how the CNS environment can be modified to become more permissive to regeneration. Axons that regenerate must find an appropriate target, but it may not be essential to reestablish the precise topography for some functions to be restored. There are now new and promising strategies for delivery of therapeutic genes to protect neurons and to stimulate regeneration. The ability to engineer cells by gene therapy combines the therapeutic values of cell transplantation and gene delivery. These remarkable developments from many disciplines have generated a new level of optimism in the search for a cure for CNS injury and in particular spinal cord injury. In this review, the authors summarize recent progress in these strategies and some of the challenges that remain in elucidating the most efficacious protocols for rescuing injured neurons, encouraging regeneration of their axons, and promoting recovery of function. PMID- 11486343 TI - Sodium channel beta subunits: anything but auxiliary. AB - Voltage-gated sodium channels are glycoprotein complexes responsible for initiation and propagation of action potentials in excitable cells such as central and peripheral neurons, cardiac and skeletal muscle myocytes, and neuroendocrine cells. Mammalian sodium channels are heterotrimers, composed of a central, pore-forming alpha subunit and two auxiliary beta subunits. The alpha subunits form a gene family with at least 10 members. Mutations in alpha subunit genes have been linked to paroxysmal disorders such as epilepsy, long QT syndrome, and hyperkalemic periodic paralysis in humans, and motor endplate disease and cerebellar ataxia in mice. Three genes encode sodium channel beta subunits with at least one alternative splice product. A mutation in the beta 1 subunit gene has been linked to generalized epilepsy with febrile seizures plus type 1 (GEFS + 1) in a human family with this disease. Sodium channel beta subunits are multifunctional. They modulate channel gating and regulate the level of channel expression at the plasma membrane. More recently, they have been shown to function as cell adhesion molecules in terms of interaction with extracellular matrix, regulation of cell migration, cellular aggregation, and interaction with the cytoskeleton. Structure-function studies have resulted in the preliminary assignment of functional domains in the beta 1 subunit. A sodium channel signaling complex is proposed that involves beta subunits as channel modulators as well as cell adhesion molecules, other cell adhesion molecules such as neurofascin and contactin, RPTP beta, and extracellular matrix molecules such as tenascin. PMID- 11486344 TI - Mechanisms of axonal plasticity: lessons from the olfactory pathway. AB - The olfactory pathway has emerged recently as an effective model for studying general principles of axon extension and regeneration. A variety of both trophic as well as repulsive molecules are found in the olfactory pathway and are being characterized for their roles in promoting the high capacity for plasticity and growth in olfactory receptor cell axons. In addition, olfactory ensheathing cells, which line the olfactory nerve, have been shown to promote axon extension not only in the olfactory pathway but also in the injured spinal cord. This review summarizes some of our current knowledge of these mechanisms and how they may function collectively to promote axon plasticity. PMID- 11486345 TI - MALDI-TOF-MS analysis of protein and DNA. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has become a powerful and widespread analytical tool in all fields of life science. The wide mass range (1-300 kDa), high accuracy, and sensitivity make it a superior method for analysis of all kinds of biomolecules including proteins, nucleic acids, and carbohydrates. In combination with 2D gelelectrophoresis, MALDI-TOF-MS is particularly suitable for the identification of protein spots via mass fingerprint or microsequencing. Furthermore, the method allows a detailed analysis of posttranslational protein modifications. Recently, the method was also successfully applied to DNA sequencing as well as screening for mutations. Thus, high-throughput genotyping of single nucleotide polymorphisms has the potential to become a routine method for both laboratory and clinical applications. PMID- 11486347 TI - The inherited episodic ataxias: how well do we understand the disease mechanisms? AB - The past few years have seen the elucidation of several neurological diseases caused by inherited mutations of ion channels. In contrast to many other types of genetic disorders, the "channelopathies" can be studied with high precision by applying electrophysiological methods. This review evaluates the success of this approach in explaining the mechanisms of two forms of episodic ataxia that are known to be caused by mutations of ion channels: episodic ataxia type 1 (EA1, caused by K+ channel mutations) and episodic ataxia type 2 (EA2, caused by Ca2+ channel mutations). Although both of these disorders are rare, they raise many important questions about the roles of identified channels in brain function. Indeed, a resolution of the mechanisms by which both diseases occur will represent a major milestone in understanding diseases of the CNS, in addition to opening the way to novel possible treatments. PMID- 11486346 TI - Functional MRI and its applications to the clinical neurosciences. AB - Functional magnetic resonance imaging (fMRI) is an emerging methodology for studying regional brain function in vivo at relatively high spatial and temporal resolution. Because MRI methods are comparatively inexpensive and entirely noninvasive, fMRI has rapidly become one of the most popular approaches for brain mapping in cognitive and systems neuroscience. There has also been great interest in using fMRI to assist in clinical diagnosis and management, with promising demonstrations of feasibility in a number of applications. Both resting and task specific regional brain activity can be measured, primarily utilizing alterations in regional cerebral blood flow (CBF) as a surrogate marker for neural function. This article reviews the biophysical and physiological bases of fMRI and its applications to the clinical neurosciences, with particular attention to potential challenges of fMRI under pathophysiological conditions. Carefully controlled prospective evaluation of clinical fMRI in its various potential applications will be required for fMRI to be validated as a clinically useful tool. Because the technology for fMRI is widely available, its impact could be substantial. PMID- 11486348 TI - The role of fine needle aspiration biopsy in the diagnosis and management of osteosarcoma. AB - We retrospectively reviewed our experience with fine needle aspiration biopsy (FNAB) in the diagnosis and management of skeletal osteosarcoma. The bi institutional study sample involved 30 consecutive aspirates from 29 patients (28 primary tumors, 1 pulmonary metastasis, 1 local recurrence). There were 17 children and 12 adults. Two aspirates were unsatisfactory for diagnosis. Of the adequate primary osteosarcoma cases analyzed by FNAB, 24 of 26 were diagnosed as osteosarcoma. All pediatric cases were correctly interpreted as osteosarcoma and treated appropriately. There were 2 incomplete diagnoses. A secondary osteosarcoma arising within an otherwise clinically, radiologically, and histologically typical giant cell tumor (malignant giant cell tumor) was not diagnosed preoperatively on FNAB due to nonrepresentative sampling. Chronologically, the first patient with osteosarcoma analyzed by FNAB was diagnosed simply as "spindle cell neoplasm." No complications resulted from the procedure. With adequate clinical and radiologic correlation, FNAB represents a technically, easily performed, cost-effective, and accurate procedure for establishing the diagnosis of skeletal osteosarcoma. Immediate interpretation of aspirated material allows for therapy planning and oncologic consultation at the initial clinic visit. PMID- 11486349 TI - CD8+ T-lymphocytes infiltrate the myocardium in fulminant herpes virus myocarditis. AB - We report two cases of fulminant viral myocarditis in previously healthy children. They were caused by herpes simplex virus (HSV)-1 (in a boy aged 3 years) and Epstein-Barr virus (EBV) (in a boy aged 12 months). We obtained the diagnosis of HSV-1 myocarditis by immunohistochemistry and the diagnosis of EBV myocarditis by in situ hybridization. Histologic examination of heart tissue from the two boys revealed mononuclear cell infiltration of the myocardium. Immunohistochemical staining identified these cells as CD8+ T-lymphocytes. CD8+ T lymphocytes induced by herpes virus infections may play an important role in the damage to heart muscle fibers seen in fulminant myocarditis in previously healthy children. To our knowledge, this is the first report of HSV-1 or EBV myocarditis (at least in children) in which viral infection has been demonstrated in the myocardium. PMID- 11486350 TI - Extraskeletical malformations in the Jarcho-Levin syndrome: postmortem study of three cases. AB - We report on pathologic examination of three autopsied newborns with Jarcho-Levin syndrome. The most important abnormalities noted were multiple extraskeletical defects such as cardiovascular, urogenital, respiratory, and central nervous system malformations. These cases add new information to the multiple vertebral segmentation defects found in this syndrome. PMID- 11486351 TI - Association between multiple intestinal atresia and omphalocele: a case report. AB - Multiple intestinal atresia is a rare disorder with vascular or hereditary etiology. So far, the occurrence of this malformation along with omphalocele has not been reported. We describe a boy born from a nonconsanguineous gypsy couple with intrauterine growth retardation, omphalocele, and multiple intestinal atresia from the pylorus to the rectum. The microscopic examination of the intestine shows multiple small lumina with a sieve-like appearance. This is characteristic of the hereditary atresias and suggests development of a defect in (re)canalization during embryogenesis. The association with omphalocele indicates a common developmental defect may be present. PMID- 11486352 TI - Clinical pathology case: hyperimmunoglobulin E syndrome (Job's syndrome). PMID- 11486353 TI - Pathology teach and tell: perinatal recessive polycystic kidney disease with congenital hepatic fibrosis. PMID- 11486354 TI - Pathology teach and tell: neonatal Marfan syndrome. PMID- 11486355 TI - Pathology teach and tell: chronic-onset hereditary tyrosinemia type I. PMID- 11486356 TI - Pathology teach and tell: congenital hepatic fibrosis and autosomal dominant polycystic kidney disease. PMID- 11486357 TI - X-ray birefringence and dichroism obtained from magnetic materials. AB - In the past decade, synchrotron radiation has triggered a surge in studies of the polarization dependence of X-ray beams passing through non-isotropic materials. A vast range of experimental results concerning polarization-dependent absorption (dichroism) and dispersion (birefringence, for example) are available from materials which are either magnetic or exhibit preferred directions due to the local atomic environment. This article aims to bring together the diversity of modern experiments in this field with established methods of optical calculus, in a way that highlights the simplicity of the underlying physics. A useful framework is formed when observable quantities, in the X-ray case, are related to atomic variables of the sample material. Atomic descriptions of absorption spectra with various levels of complexity are considered, and some well documented sum-rules are encountered. The framework is the most general allowed within the electric dipole approximation. By way of illustration, dichroic X-ray absorption by two materials with highly anisotropic properties and magnetic ions with different valence shells are considered; namely, a 3d-transition ion in ferrous niobate, and a lanthanide ion in dysprosium borocarbide. Both materials display interesting magnetic properties that are challenging to interpret at an atomic level of detail, and it is shown how absorption experiments can contribute to resolving some issues. PMID- 11486358 TI - Use of highly energetic (116 keV) synchrotron radiation for X-ray fluorescence analysis of trace rare-earth and heavy elements. AB - This study has revealed the advantages of the use of 116 keV X-rays as an excitation source of X-ray fluorescence (XRF) analyses. This technique is suitable for nondestructive multielemental analyses of heavy elements such as rare-earth elements. The lowest MDL value evaluated for the bulk analysis of a JG 1 standard reference sample (granite rock) was 0.1 ppm for W for a 500 s measurement. The spectrum of standard glass samples of SRM612 demonstrated clearly resolved K-line peaks of more than 30 elements, including all the existing rare-earth elements, at 50 ppm levels. The calibration curve for the determination of a rare-earth element shows a linear relation between the XRF intensity and concentrations from 10 to 0.03 ng. This powerful technique should be useful for nondestructive analyses of rare-earth and heavy elements in geological, geochemical and archaeological samples as well as industrial materials. PMID- 11486359 TI - Crystal monochromator with a resolution beyond 10(8). AB - Monochromatization with crystal diffraction has been achieved to a resolution (lambda/delta lambda) beyond 10(8). The monchromator is specifically designed for 23.880 keV synchrotron radiation (lambda = 51.9 pm) for applications involving nuclear resonant scattering from 119Sn. The design uses asymmetrically cut silicon (12 12 12) crystal reflections from two single-crystalline monoliths oriented in a dispersive geometry. A transmitted energy bandwidth of 140 +/- 20 mu eV was measured, corresponding to a resolution of 1.7 x 10(8). Methods of improving efficiency, wavelength stability and resolution are discussed. PMID- 11486360 TI - Geometrical fitting of experimental XANES spectra by a full multiple-scattering procedure. AB - In this paper a new software procedure is presented, named MXAN, able to fit the XANES part (from the edge to about 200 eV) of experimental X-ray absorption data. The method is based on the fitting between the experimental spectrum and several theoretical calculations generated by changing the relevant geometrical parameters of the site around the absorbing atom. The X-ray photo-absorption cross section is calculated using the full multiple-scattering scheme; different choices of the exchange correlation part of the potential can be utilised. To show the potentialities of the method the analysis of the nickel K-edge of the nickel ion in aqueous solution is presented. The procedure is able to recover the correct information on the symmetry and atomic distances from the experimental Ni K-edge XANES spectrum. The recovered structure is found to be independent of the starting conditions, showing the theoretical uniqueness of the structural solution. PMID- 11486361 TI - An investigation of the use of the Hedin-Lundqvist exchange and correlation potential in EXAFS data analysis. AB - In real systems, inelastic processes remove photoelectrons from the elastic scattering channel. This reduces the amplitude of the EXAFS. Traditionally the discrepancies between experimental and theoretical amplitudes were treated by including two semi-empirical reduction factors in the data analysis. Some inelastic effects may, however, be modelled more rigorously using a complex exchange and correlation potential, for example the Hedin-Lundqvist (HL) potential used in most EXAFS data-analysis programs. In this paper a systematic study of the effects of the HL potential on the calculated EXAFS amplitudes is presented. Expressions are derived whereby the EXAFS amplitudes may be examined in the presence of an arbitrary complex potential independently to the rest of the EXAFS signal. These results are used to study the effects of the HL potential on EXAFS data analysis in detail. PMID- 11486362 TI - Multiple-electron excitation in X-ray absorption: a simple generic model. AB - The probability of multiple-electron excitation in X-ray absorption is calculated using a simple generic model. The model permits calculations to be made for all atoms with little input data or computing effort. The high-energy limit of this probability, which gives the usual EXAFS amplitude reduction factor, is calculated in the 'sudden approximation' using Slater orbitals. Good agreement with experiment is found. The energy dependence of this probability is also calculated using a simple model form of perturbing potential and found to agree well with experiment for rare gas atoms. The effect on the X-ray absorption coefficient of including multiple-electron excitations is also determined and is found to be small, again in agreement with observation. PMID- 11486363 TI - CRYSTALFF--from crystallography to EXAFS multiple-scattering calculations with FEFF. AB - CRYSTALFF is an alternative utility to ATOMS containing most of the features of this standard program for converting crystallographic data to FEFF input. In addition, it offers an interface with molecular modelling programs via the PDB format and new coordination sphere analysis options. PMID- 11486364 TI - Overview of the tunable beamlines for protein crystallography at the EMBL Hamburg Outstation; an analysis of current and future usage and developments. AB - The EMBL Hamburg Outstation currently operates two tunable protein crystallography beamlines suitable for single and multiple anomalous diffraction (SAD/MAD) experiments. The first beamline, designated X31, is located on a bending magnet of the DORIS III storage ring whereas the second beamline, BW7A, is positioned at a multipole wiggler at the same storage ring. X31 is equipped with an energy stabilization device to ensure constant wavelength during longer data-collection periods. The in-house built crystallographic end-station is now equipped with a Mar345 imaging-plate scanner as a detector. The wiggler beamline BW7A features a novel sagitally focusing monochromator. The end-station used here has also been developed and built in-house. The beamline is currently operated with a Mar 165 CCD detector. In this paper the hardware and software developments of the last years will be summarized and the outlook for substantial upgrades will be given. The future plans include the design and construction of a third tunable beamline, designated X12, for protein crystallography. The development of automated beamlines for protein crystallography is of particular importance with respect to structural genomics initiatives. The analysis of the projects of the last years shows the wide range of anomalous scatterer used on the tunable beamlines thus demonstrating the need of a wide range of accessible energies and fast and reliable energy changes. PMID- 11486365 TI - NEXAFS study of 1-butanethiol adsorbed on Cu(111) and square root of 7 x square root of 7 R19.1 degrees S/Cu(111). AB - The adsorption of 1-butanethiol on Cu(111) and square root of 7 x square root of 7 R19.1 degrees S/Cu(111) surfaces has been studied by S K-edge near edge X-ray absorption fine structure (NEXAFS) spectroscopy and thermal desorption spectroscopy. Upon adsorption on clean Cu(111) surface at room temperature, butanethiolate as well as atomic sulfur is formed. For the butanethiolate, the S C bond is found predominately perpendicular to the surface as revealed by polarization analysis. In contrast, on square root of 7 x square root of 7 R19.1 degrees S/Cu(111) surface, the S-H and S-C bonds of the butanethiol stay intact, resulting in a weakly chemisorbed butanethiol. PMID- 11486366 TI - Detailed tabulation of atomic form factors, photoelectric absorption and scattering cross section, and mass attenuation coefficients in the vicinity of absorption edges in the soft X-ray (Z = 30-36, Z = 60-89, E = 0.1-10 keV)- addressing convergence issues of earlier work. AB - A new tabulation of atomic form factors is discussed briefly, extending the validity of the isolated atom approximation and serving as a baseline for near edge solid-state and XAFS investigations. This is detailed by Chantler [J. Phys. Chem. Ref. Data, (2000), 29, 597-1048] and is the latest component of the FFAST tabulation of NIST. PMID- 11486367 TI - [An ABC transporter mediating the membrane detachment of lipid-modified proteins]. PMID- 11486368 TI - [Blue-light receptors in plants]. PMID- 11486369 TI - [Refolding of proteins from inclusion bodies]. PMID- 11486370 TI - [Topology and recognition function of glycolipids]. PMID- 11486371 TI - [Structural biology of heparan sulfate sulfotransferase]. PMID- 11486372 TI - [A local structure assisting the enzyme catalysis from behind the catalytic center]. PMID- 11486373 TI - [Crystal structure of the large ribosomal subunit]. PMID- 11486374 TI - [The present situation and significance in mouse genome sequencing]. PMID- 11486375 TI - [Degradation of thiometon in ethyl acetate]. AB - When performing multiresidue analysis of pesticides, the recovery of thiometon was less than 20% from carrots and eggplants, but about 100% from garlic chives and welsh onions. The recovery of thiometon was found to depend on the lot of ethyl acetate. A 2-year-old lot of ethyl acetate caused degradation of thiometon, but a fresh lot of ethyl acetate did not. Analysis showed that ethyl acetate stored for 2 years contained about 5 microL/mL of acetaldehyde. Thiometon was also degraded by acetone or acetonitrile, when acetaldehyde was added to them, in the same manner as by aged ethyl acetate. The fact that the recovery of thiometon from welsh onions was about 100% indicated that some of the mercaptans in allium vegetables may prevent thiometon degradation. Mercaptans such as L-cysteine and 3 mercaptoproionic acid were confirmed to prevent the degradation of thiometon and disulfoton. These findings show that mercaptans may be useful additives for analyzing thiometon and disulfoton. PMID- 11486376 TI - [Analysis of main pigments and other ingredients in lac color product]. AB - The contents of the main pigments and other ingredients in commercial lac color products were determined by HPLC using an RP-18 column with 0.1 mol/L citric acid buffer solution-methanol (16:5) as the mobile phase, and a photodiode array (PDA) detector set at 280 nm and 490 nm. The main pigments were confirmed by PDA and electrospray ionization mass spectrometry. Laccaic acids A, B, C and E were detected in all lac color products, and the ratio of content of laccaic acid A in all products was over 50%. The total contents of laccaic acids A, B and C in lac color food additive products and reagent products were 775-858, 797 and 779 g/kg, respectively. As for the contents of ingredients except pigments in commercial food additive products, the maximum moisture content was about 10%, and ether soluble substances amounted to 0.5-3.6%. PMID- 11486377 TI - [Modification of HPLC conditions for the determinations of raw materials, intermediates and subsidiary colors in 5 kinds of food azo colors]. AB - Modification of HPLC conditions for the determinations of raw materials, intermediates and subsidiary colors (organic impurities) in 5 kinds of food azo colors were studied in order to analyze them simply and rapidly. The organic impurities were determined by HPLC using L-column ODS and a gradient system (0.02 mol/L ammonium acetate solution for 10 min, and a gradient with a mixture of acetonitrile and water (7:3)). The organic impurities in 163 samples of azo colors subjected to inspection in fiscal year 1999 were determined under the proposed HPLC conditions. It was found that the contents of organic impurities in 163 samples were less than the limit of Japan's Specifications and Standards for Food Additives, 7th Edition. PMID- 11486378 TI - [Daily intake of isoflavones based on the market basket method]. AB - Daily intake of isoflavones (daidzin, glycitin, genistin, daidzein, glycitein, and genistein) was determined quantitatively, based on the market basket method. Acid hydrolysis during extraction of foods was chosen to convert phytoestrogenes into the respective aglycons, facilitating HPLC analysis and allowing quantitation of total isoflavones as aglycones including both originally present glycosides and "free" aglycones. The isoflavones were extracted from samples with methanol and determined by reversed-phase HPLC analysis using a linear gradient of methanol-water as the eluent. From the results of hydrolysis, the daily intake of total isoflavon was 38.1 mg/adult Japanese. The values obtained by the market basket method and the National Nutrition Survey method were similar. PMID- 11486379 TI - [Contents of eleven phthalates and di(2-ethylhexyl) adipate in retail packed lunches after prohibition of DEHP-containing PVC gloves for cooking purposes]. AB - Ten samples of retail packed lunches purchased from convenience stores were determined for 11 phthalates and di(2-ethylhexyl) adipate (DEHA) in August 2000, 2 months after the prohibition of DEHP-containing PVC gloves in Japan. Each homogenized sample was extracted with acetonitrile, partitioned with n-hexane, and cleaned up using Florisil and PSA columns. Phthalates in the extract were determined by GC/MS (SIM). The limits of detection were 14.9 ng/g for di(2 ethylhexyl) phthalate (DEHP) and 18.6 ng/g for dibutyl phthalate (DBP). Levels of phthalates in packed lunch samples were 45 to 517 ng DEHP/g (198 ng/g, average), ND to 90 ng DEHA/g, and ND to 10.0 ng BBP/g. Diisononyl phthalate (DINP) was detected in one sample at 76 ng/g. Average DEHP level in ten samples was 4% of that in 1999. The contents of other phthalates were also reduced. DBP was not detected in any sample. Recovery of deuterated isomers added as surrogates was 27.9% for DNP-d4, and 40.6 to 101.5% for the other phthalates. PMID- 11486380 TI - [Determination of sucralose in foods by anion-exchange chromatography and reverse phase chromatography]. AB - Simple and rapid methods for the determination of sucralose in foods were developed using anion-exchange chromatography (AEC) with pulsed amperometric detection and reverse-phase HPLC with refractive index detection. Sucralose was extracted with water or methanol, and the extracted solution was cleaned up on a Sep-Pak C18 cartridge and a Sep-Pak Alumina N cartridge. The AEC separation was performed on a CarboPac PA1 column (4.0 mm i.d x 250 mm) using 100 mmol/L sodium hydroxide-50 mmol/L sodium acetate solution as the mobile phase at a flow rate of 1.0 mL/min. The recoveries of sucralose from foods were 80.6-102.0%, and quantitation limits from foods except chewing gum were 0.5 microgram/g (2 micrograms/g from chewing gum). The reverse-phase HPLC separation was performed on an Inertsil ODS-3V column (4.6 mm i.d x 150 mm) using methanol-water (25:75) as the mobile phase at a flow rate of 1.0 mL/min. The recoveries of sucralose from foods were 80.2-121.2%, and quantitation limits from foods except chewing gum were 5 micrograms/g (20 micrograms/g from chewing gum). PMID- 11486381 TI - [Determination of sucralose in foods by HPLC]. AB - A method for the determination of sucralose in various foods by RI-HPLC and ion chromatography with a pulsed amperometric detector (PAD-IC) was developed. Chopped or homogenized samples were packed into cellulose tubing with 0.01 mol/L hydrochloric acid containing 10% sodium chloride and were dialyzed against 0.01 mol/L hydrochloric acid for 24 hours. The dialyzate was passed through a Bond Elut ENV cartridge, and the cartridge was washed with 0.2 mol/L NaOH and water. Sucralose was eluted from the cartridge with methanol. The extract was taken to dryness in an evaporator and the residue was re-dissolved in water. Sucralose was separated on an Inertsil ODS-3V column with a mobile phase of acetonitrile-water (15:85) and an RI detector. It was also determined on a CarboPak PA1 column with a mobile phase of 100 mmol/L NaOH-75 mmol/L CH3COONa, using a PAD detector. The recoveries of sucralose from various kinds of foods spiked at 50 micrograms/g and 200 micrograms/g ranged from 88-105%. The detection limit in samples was 10 micrograms/g for RI-HPLC and 1 microgram/g for PAD-IC. PMID- 11486382 TI - [Antigenicity and phototoxicity of water-soluble extract from Salacia reticulata (Celastraceae)]. AB - The antigenicity and phototoxicity of water-soluble extract from Salacia reticulata (SRE) were examined in guinea pigs. In a study of active systemic anaphylaxis reaction, neither the oral administration group (64 or 320 mg/kg, 5 times/week, 3 weeks) nor the subcutaneous administration group (64 mg/kg, 1 time/week, 3 weeks) exhibited any anaphylactic reaction. Moreover, sensitization with serum obtained from these animals did not induce passive cutaneous anaphylaxis reaction in normal animals. In a phototoxicity study, oral administration of SRE (320 mg/kg) induced neither erythema nor edema. These results suggest that SRE is not antigenic or phototoxic. PMID- 11486383 TI - [Drying ability of anhydrous sodium sulfate on wet organic solvents after liquid liquid partition]. AB - Water concentration in organic solvents after liquid-liquid partition was determined by the Karl Fischer titration method. n-Hexane and petroleum ether showed quite low levels of water, such as 0.1 mg/mL. The water concentration in wet ethyl acetate was about 20-30 mg/mL and that in diethyl ether was about 8-10 mg/mL. Anhydrous sodium sulfate absorbed about 20-25% of the water after vigorous mixing with wet ethyl acetate or diethyl ether. Wet acetonitrile extract from wet food, which contained about 60 mg/mL water after salting out with sodium chloride, was not dried at all with anhyfrous sodium sulfate treatment. Spiking n hexane into wet ethyl acetate or wet diethyl ether was effective to exclude water. Spiking toluene into salted acetonitrile drove out water and dissolved sodium chloride. It can be concluded that the drying ability of anhydrous sodium sulfate towards wet organic solvents is poor, but it is effective in removing suspended water in solvents. PMID- 11486384 TI - [The fate of dioxins during green tea manufacture]. AB - Dioxin concentrations and homologue profiles were examined in plucked new shoots, crude tea and its hot water extracts, soils and atmosphere in tea orchards. The rate of dry provisions in crude tea has increased 4 times as that of the plucked new shoots. However, dioxin concentrations except O8CDD in crude tea have increased only 2 to 3 times as those of the plucked new shoots. O8CDD concentration increased remarkably during processing. Little dioxins were detected in hot water extracts of crude tea leaves. PMID- 11486385 TI - [Antioxidative compounds in spices and herbs]. PMID- 11486386 TI - [Determination of bisphenol A in foods using GC/MS]. AB - An analytical method using GC/MS was developed for bisphenol A (BPA) in foods and BPA was determined in canned foods and fresh foods such as vegetables, fruit and meat. BPA was extracted with acetone from the samples and the extract was concentrated at under 40 degrees C in vacuo to afford an aqueous solution, which was washed with hexane after alkalization and extracted with 50% diethyl ether hexane after acidification. Extracts were cleaned up on a PSA and/or a C18 cartridge column, and BPA was derivatized with heptafluorobutyric anhydride and determined by GC/MS (SIM). This method was applicable to the detection and determination of BPA residues in food samples at the level of 1 ng/g. Among canned foods, BPA was found in 6 corned beef, 1 chicken, 9 sweet corn and 3 bean samples at the levels of 17-602 ng/g, 212 ng/g, 2.3-75 ng/g and 3.5-26 ng/g, respectively. BPA was also detected in 1 retort soup and 1 retort pack product at the levels of 11 ng/g and 86 ng/g, respectively. As for dairy products, BPA was not detected in butter and milk. Among fresh foods, BPA was detected in 2 fish and 3 liver samples at the levels of trace (tr)-6.2 ng/g and tr-2.2 ng/g, respectively. In vegetables, fruits and chocolates, a trace level of BPA was detected in only 1 chocolate. Traces of BPA were also detected in 3 samples of 6 boxed lunches. PMID- 11486387 TI - Methods for determination of milt protein and epsilon-polylysine in food additive preparations and processed foods by capillary zone electrophoresis. AB - A simple and rapid method using capillary zone electrophoresis (CZE) for the determination of milt protein (MP), which contains mainly protamine, and polylysine (PL) in food additive preparations and processed foods was developed. CZE separation was performed on poly(vinyl alcohol)-coated capillaries at a column temperature of 20 degrees C with 120 mmol/L phosphate buffer (pH 2.5) as the running buffer. The influence of various components in food additive preparations on CZE analysis of MP and PL was examined. Egg white lysozyme, glycine, sodium acetate, glycerol, fumaric acid, calcium carbonate, dextrin, emulsifiers and sodium polyphosphate and pyrophosphate had no effect. No peak of protamine was detected in preparations containing metaphosphate. The analysis method for processed foods was composed of extraction with 4% formic acid, precipitation of macromolecular compounds with ethanol, concentration in a water bath and determination by CZE. The average recoveries were 108.4% for protamine sulfate (PS) in red bean sticky rice, and 81.3% for PL in white rice, 118% in egg sandwiches, and 115% in shiraae. The limits of detection of PS in red bean sticky rice and PL in white rice were both 50 ppm. PMID- 11486388 TI - Analytical method for Buddleja colorants in foods. AB - Buddleja yellow colorant derived from Buddleja officinalis Maxim. has recently been approved for use as a new kind of natural colorant for food additives in China. In order to distinguish Buddleja yellow colorant from other yellow colorants, two known phenylpropanoid glycosides, acteoside (= verbascoside) and poliumoside, were isolated from the colorant as marker substances for Buddleja yellow colorant. Poliumoside has not been detected in B. officinalis Maxim. previously. These phenylpropanoid glycosides were not detected in the fruits of Gardenia jasminoides Ellis or in the stamens of the flowers of Crocus sativus L., which also contain crocetin derivatives as coloring components, using a photodiode array and mass chromatograms. Thus, an analytical HPLC method was developed to distinguish foods that have been colored with yellow colorants containing crocetin derivatives, using phenylpropanoid glycosides as markers. PMID- 11486389 TI - [Determination method of polysorbates in powdered soup by HPLC]. AB - A method for qualitative and quantitative analyses of polysorbates in powdered soup by HPLC was studied. Polysorbates in samples were extracted with acetonitrile after rinsing with n-hexane to remove fats and oils. The extract was cleaned up using a Bond Elut silica gel cartridge (500 mg). The cartridge was washed with ethyl acetate and polysorbates were eluted with a small amount of acetonitrile-methanol (1:2) mixture. The eluate was treated with cobalt thiocyanate solution to form a blue complex with polysorbate. In order to determine polysorbate, the complex was subjected to HPLC with a GPC column, using a mixture of acetonitrile-water (95:5) as a mobile phase, with a detection wavelength of 620 nm. The recoveries of polysorbate 80 added to powdered soups were more than 75% and the determination limit was 0.04 mg/g. When the proposed method was applied to the determination of polysorbates in 16 commercial samples of powdered soup for instant noodles and seasoning consomme, no polysorbates were detected in any sample. PMID- 11486390 TI - [Fukuronori (Gloiopeltis furcata) extract: 90-day dietary toxicity study in F344 rats]. AB - Fukuronori extract (FE), which is mainly composed of polysaccharides, and is an extract of the seaweed Gloiopeltis furcata, is permitted for use as a food thickening agent by the Ministry of Health and Welfare, Japan. In order to study the subchronic toxicity of FE, F344 rats of both genders were administered FE at concentrations of 0% (basal diet, control group), 0.5%, 1.5% and 5.0% in basal powder diet for 90 days, and observation of general condition, recording of body weight and food consumption, examination of hematology and blood chemistry, measurement of organ weight, and pathological examination were performed. Food consumption tended to increase in both sexes given FE at 1.5% and 5.0% throughout most of the experimental period. This was, however, considered not to be a toxic effect because the differences in body weight were small. Total cholesterol and triglycerides in serum decreased significantly (p < 0.05) and not significantly, respectively, in males of the 5.0% group. These changes were considered to be related to the intake of FE, but the differences were slight and within physiological ranges. Hematological and pathological examination revealed neither any particular adverse effect nor any significant difference from the control. Hence, dietary intake of 5.0% of FE, 3,362 mg/kg/day for males and 3,594 mg/kg/day for females as mean daily intake, for 90 days was considered to be a no observable adverse effect level in rats. PMID- 11486391 TI - [Unique identification system for the safety assessment of genetically modified organisms; unit and transparency]. PMID- 11486392 TI - [Principles of PCR technologies]. PMID- 11486393 TI - [An introduction to enantiomeric purity determination]. PMID- 11486394 TI - The role of ifosfamide in cytoreduction, stem cell mobilization and high-dose therapy in relapsed/refractory malignant lymphomas. PMID- 11486395 TI - Combination chemotherapy containing mitoguazone, ifosfamide, methotrexate, etoposide (MIME) and G-CSF efficiently mobilize peripheral blood progenitor cells in heavily pre-treated relapsed lymphoma patients. AB - In this study we explored whether a standard chemotherapy regimen consisting of mitoguazone, ifosfamide, methotrexate and etoposide (MIME) combined with 5 micrograms/kg or 10 micrograms/kg G-CSF was capable of mobilizing peripheral blood progenitor cells (PBPC) in lymphoma patients. Thirty-three patients with Hodgkin's disease (HD) and 108 patients with non-Hodgkin's lymphoma (NHL) were mobilized with MIME/G-CSF. Most patients were heavily treated with different chemotherapy regimens receiving a median of 11 cycles (range 3-40) of chemotherapy prior to mobilization. Eight of 141 patients failed to mobilize PBPC and bone marrow was harvested. In addition, 10 patients obtained a harvest of < 2.0 x 10(6) CD34+ cells/kg. More than 2.0 x 10(6) CD34+ cells/kg were achieved in all HD patients and in 83% of the NHL patients. Fifty-eight per cent of the patients harvested > or = 5 x 10(6) CD34+ cells/kg. Eleven per cent of the patients developed neutropenic fever during the mobilization and 3% had nadir platelet values below 20 x 10(9)/L. An inverse correlation was observed in high grade NHL (H-NHL) patients between the number of chemotherapy cycles given before mobilization and yield of CD34+ cells. Such an association was not seen among patients with HD, transformed and low-grade NHL. When G-CSF 10 micrograms/kg was used in combination with MIME, this correlation was no longer seen in patients with H-NHL. There was also association between CD34+ cell yield and prior radiotherapy in patients with HD or transformed NHL or low-grade NHL. These results demonstrate that an ordinary salvage chemotherapy regimen, such as MIME combined with G-CSF, can be successfully used to mobilize PBPC. Although no significant effect of increased dose of G-CSF was found, our data suggest that MIME/G-CSF 10 micrograms/kg should preferentially be used to mobilize PBPC in H NHL patients pre-treated with > or = 12 cycles of chemotherapy, in irradiated HD patients and in all low-grade and transformed lymphomas. PMID- 11486396 TI - Ifosfamide- and etoposide-based chemotherapy as salvage and mobilizing regimens for poor prognosis lymphoma. AB - Treatment of early relapsing or resistant non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD) remains problematic. High-dose chemotherapy followed by autologous peripheral blood stem cell (PBSC) transplantation improves the prognosis for patients in response following standard dose regimens. We adopted the strategy of using salvage chemotherapy to debulk disease and simultaneously mobilize stem cells. We used regimens based on ifosfamide and etoposide because these drugs are not usually used as the front-line treatment. Twenty-seven patients with NHL received MINE chemotherapy (mesna and ifosfamide 1330 mg/m2 and etoposide 65 mg/m2 i.v. days 1-3, and mitoxantrone 8 mg/m2 i.v. day 1). The same schedule, but higher doses were used for PBSC stimulation (mesna, ifosfamide 1700 mg/m2, etoposide 175 mg/m2, mitoxantrone 10 mg/m2). Forty-six patients with HD received VIM chemotherapy (mesna, ifosfamide 1200 mg/m2 i.v. days 1-5, etoposide 90 mg/m2 i.v. days 1, 3, and 5, methotrexate 30 mg/m2 i.v. days 1 and 5). After both VIM and high dose MINE, chemotherapy for mobilization was followed by G-CSF administered at a dose 5-16 micrograms/kg/day depending on the clinicians judgement of the patient's pretreatment. Complete response after VIM and MINE were 39% and 38%, respectively; partial response (PR) rates were 17% and 29%, and stable disease (SD) 25% and 4%, respectively. In both groups, patients with relapsing disease had better responses than did those with primary progressive disease. Both regimens exhibited excellent mobilizing capacity. We performed 213 aphereses with a median 3 per patient starting on either day 13 as a median for VIM, or on day 12 as a median for MINE. In the majority of patients, the collection started in the time interval median +/- 1 day (n = 62, 85%). The median yields were 10.6 x 10(6) CD34+ cells/kg and 53.1 x 10(4) CFU-GM/kg for VIM, and 13.3 x 10(6) CD34+ cells/kg and 54.5 x 10(4) CFU-GM/kg for MINE. We collected at least 2.5 x 10(6) CD34+ cells/kg in all but six patients (8%), and the harvested amount of CD34+ cells was less than 1.0 x 10(6)/kg in only two patients (3%). The toxicity of VIM and MINE was minimal and chemotherapy-induced trombocytopenia did not occur with PBSC collection. PMID- 11486397 TI - High-dose ifosfamide in combination with etoposide and epirubicin (IVE) in the treatment of relapsed/refractory Hodgkin's disease and non-Hodgkin's lymphoma: a report on toxicity and efficacy. AB - One hundred and seven patients (61 with diffuse large B-cell non-Hodgkin's lymphomas and 46 with Hodgkin's disease) in relapse or following of primary therapy received ifosfamide 3 g/m2 i.v. daily for 3 days in combination with epirubicin 50 mg/m2 i.v. day 1 and etoposide 200 mg/m2 i.v. days 1-3. Of the 46 patients with Hodgkin's disease (28 male, 18 female, and a median age of 28 years) 85% of patients had a response to treatment, with 17 achieving complete remission and 11 good partial remission. Twenty-eight proceeded to autologous bone marrow or peripheral blood stem cell transplantation. Twenty-three patients remain alive in continuous remission with a follow-up of 12-61 months. The median overall survival time for all patients in this group is 36 months. Haematological toxicity, particularly WHO Grade IV neutropenia, occurred in all patients but improved over the three courses of treatment. There was no major non haematological toxicity. Further trials of this regimen in this clinical situation are indicated. The patients with non-Hodgkin's lymphomas in this study had diffuse large B-cell lymphomas and had only received first-line treatment. Twenty had primarily refractory disease, 15 had only achieved partial remissions (PR), and 26 had developed relapse following primary treatment. The overall response rate was 43%; it was 60% for those who had achieved initial PR, 58% for those in relapse after an initial CR or very good PR following initial therapy, but only 10% for those with primarily refractory disease. Tolerance to the regimen was similar to that observed in treatment of the patients with Hodgkin's disease and many were able to undergo stem cell collection, following mobilization with this regimen. The 2-year overall survival result was 22% for patients with some response to first-line treatment but 0% for primary refractory patients. PMID- 11486398 TI - High-dose treatment with autologous stem cell transplantation versus sequential chemotherapy: the GELA experience. AB - Autologous stem-cell transplantation (ASCT) has permitted to deliver high-dose therapy (HDT). In aggressive lymphomas, the GELA group conducted prospective and retrospective studies comparing HDT + ASCT to conventional sequential chemotherapy. In relapsing patients and in partial remission, retrospective studies showed a survival advantage for HDT + ASCT over sequential chemotherapy. In complete response, advantage for HDT + ASCT was demonstrated in a prospective trial only for patients with high intermediate or high risk in the IPI score. The attainment of a maximal reduction of the tumoral mass before going HDT is very important either in first line or in relapsing patients. PMID- 11486399 TI - Effective progenitor cell mobilization in lymphoproliferative disorders using ifosfamide, epirubicin and etoposide (IEV). AB - The combination of ifosfamide, epirubicin and etoposide (IEV) is an effective salvage regimen for lymphoproliferative disease. We report our experience with this combination in mobilization of peripheral blood stem cells (PBSC) in patients with relapsed or refractory/high-risk lymphoma. The median time to leukapheresis was 14 days, with 85% of patients commencing PBSC collection in the range of 13-15 days. Mobilization was successful in 26 of 28 patients (93%), who achieved the minimum transplant dose of 2 x 10(6)/kg CD34+ cells in a median of 2 leukaphereses. Overall, the median CD34+ cell yield was 6.94 x 10(6)/kg (range 0.73-27.4). In 15 of 27 patients (54%), the yield was sufficient (> 6 x 10(6)/kg) to permit CD34+ cell selection and/or a second autograft. IEV was given as an inpatient in all cases. Patients were scheduled for discharge after chemotherapy. This was achieved in 71%, with readmission 1 week later for harvest. Therapy was complicated by neutropenic fever in 13 patients and mild nausea. In autografts carried out using IEV-mobilized PBSC (n = 20), the median time to neutrophils > 0.5 x 10(9)/L was 10 days (range 7-13 days), and to platelets > 20 x 10(9)/L was 13 days (range 11-18 days). There was no mobilization- or transplant-related mortality. We conclude that IEV is a safe, predictable and highly effective mobilization regimen in patients with lymphoma. PMID- 11486400 TI - The role of ifosfamide in the treatment of relapsed and refractory lymphoma. AB - Ifosfamide is an alkylating agent with proven efficacy in the treatment of solid tumours and malignant lymphomas. Because it causes only mild to moderate myelosuppression, ifosfamide is often used in combination regimens with other agents. Ifosfamide has been mainly used in therapy of lymphoma as a component of salvage regimens, but high-dose ifosfamide is also effective in the mobilization of peripheral stem cells for treatment of patients with relapsed or refractory lymphoma with regimens containing autologous stem cell transplantation. Based on promising data with a new combination regimen containing idarubicin, etoposide and ifosfamide (IIVP-16) in patients with poor-risk non-Hodgkin's lymphoma, we have performed a phase II study using DIZE (dexamethasone 20 mg i.v. days 1-4, idarubicin 8 mg/m2 i.v. days 1 + 2, ifosfamide 1.0 g/m2 continuous infusion (c.i.) days 1-4, and etoposide 160 mg/m2 c.i. days 1-4) in patients with relapsed or refractory Hodgkin's and non-Hodgkin's lymphoma. In 43 evaluable patients, the response rate was 58%, including 11 complete remissions (CR) and 14 partial remissions (PR). The mean duration of response was 8 months (1-30). Myelosuppression was generally mild with mean duration of neutropenia < 1000/microL of 2.5 days (range 0-18) and thrombocytopenia < 25,000/microL of 1.5 days (0-17). Thus, DIZE is an effective and safe regimen for pretreated patients with aggressive lymphoma. These results appear to compare favourably with other salvage regimens such as IMVP-16 or DHAP. In conclusion, salvage regimens containing ifosfamide can play an important role in patients who are not eligible for high-dose chemotherapies. Moreover, ifosfamide might also have a role in reducing tumour burden and selecting those patients who qualify for HDCT. PMID- 11486401 TI - DICE (dexamethasone, ifosfamide, cisplatin, etoposide) infusional chemotherapy for refractory or relapsed non-Hodgkin's lymphoma (NHL). AB - Continuous exposure to naturally-derived chemotherapy agents such as etoposide may theoretically override drug resistance due to overexpression of the multidrug resistance gene product, p-glycoprotein. Dexamethasone in high dose may have a similar overriding effect. Data also suggest that ifosfamide both by continuous infusion and when combined with a platinum compound may be more effective. Forty four chemotherapy-refractory/relapsed lymphoma patients received the DICE infusional regimen. The programme consisted of dexamethasone 40 mg i.v. daily for 4 days, ifosfamide 1500 mg mixed with equal dosing of mesna continuously infused i.v. daily for 4 days, cisplatin 25 mg i.v. each day for 4 days and etoposide 150 mg continuously infused i.v. daily for 4 days, all administered every 3 weeks. Doses of ifosfamide and etoposide were escalated by 250 mg and 25 mg, respectively, based on patient tolerance, usually lack of myelosuppression. Special hydration was not required. G-CSF support was provided to patients as required. All patients had disease that had relapsed from or was resistant to CHOP or a similar anthracycline-containing combination regimen. A majority had previously received at least two regimens and 27% had received three or more. Of 44 patients, 32 (73%) achieved a significant response consisting of 18 complete remissions (CR) (41%) and 14 (32%) partial remissions (PR). There were 81% objective responses in large cell lymphoma comprised of 50% CR and 31% PR. Previous response to chemotherapy predicted response to DICE: 83% (25/30) of prior responders vs 50% (7/14) of non-responders had a response to the treatment regimen (p = 0.031, Fisher's exact test). Patients not undergoing transplantation had a median time of 8 months on therapy and a mean of 10 months. Toxicity was haematological (36% developing grade III-IV toxicity) and neurological (9%). There were only three episodes of clinical cystitis or gross haematuria. Infusional DICE is an easily administered and well tolerated programme with significant activity in refractory or relapsed NHL and may be useful as a tumour reductive therapy prior to high-dose chemotherapy and autologous stem cell transplantation. PMID- 11486402 TI - ICE--an efficient drug combination for stem cell mobilization and high-dose treatment of malignant lymphoma. AB - Between 1989 and 1999 we studied the ICE regimen in sequential trials in 290 patients with malignant lymphoma and germ-cell tumours. For patients with relapsed or refractory lymphoma we could demonstrate a comparable efficacy of ICE to other high-dose chemotherapy (HDCT) regimens but with a toxicity profile in favour of ICE. From a retrospective comparative analysis of ICE as HDCT regimen in patients with malignant lymphoma and germ-cell tumours we conclude that the characteristic toxicity profile of ICE varies depending on prior drug exposure of individual patients. Further dose intensification of ICE may be achieved with acceptable toxicity by adding further drugs (e.g. anthracyclines) or by treatment with sequential cycles of ICE (Tandem-HDCT). More convenient drug formulations (e.g. etoposide phosphate) might further improve the therapeutic index of ICE. PMID- 11486403 TI - High-dose ifosfamide and vinorelbine as salvage therapy for relapsed or refractory Hodgkin's disease. AB - In an effort to improve results in patients with relapsed or refractory Hodgkin's disease (HD), an intensive regimen combining vinorelbine (25 mg/m2 i.v. days 1 and 5) and high-doses of ifosfamide (3000 mg/m2/d, days 1-4 by continuous infusion) with mesna uroprotection and G-CSF support was designed. Forty-seven patients were treated; 14 had failure to initial induction therapy and 33 had disease relapsed from an initial response. The response rate was 83%, with 21 complete (45%, CR) and 18 partial remissions (38%, PR). Partial response was achieved after a median of two cycles (range 1-3) and CR after a median of six cycles (range 2-10). At the end of ifosfamide and vinorelbine, 10 patients in CR, one in PR, and one with stable disease also received radiotherapy to nodal sites of relapse. Eleven patients who had undergone peripheral blood stem cell (PBSC) harvesting following ifosfamide-vinorelbine proceeded to receive high-dose chemotherapy (HDCT) and PBSC transplantation. The main toxic effect was grade III IV neutropenia, documented in 65% of cycles with a median duration of 4 days, and non-haematological toxicity was mild. The combination of high-doses of ifosfamide and vinorelbine was well tolerated and an active regimen in treatment of patients with relapsed and refractory HD. It was not only useful as salvage therapy with or without consolidative radiotherapy but it also was a valuable induction regimen before high-dose intensification therapy followed by PBSC reinfusion in patients eligible for this approach. PMID- 11486404 TI - Rituximab and ifosfamide, mitoxantrone, etoposide (RIME) with Neupogen support for B-cell non-Hodgkin's lymphoma prior to high-dose chemotherapy with autologous haematopoietic transplant. AB - A phase I/II study was performed to analyse the ability of ifosfamide-based chemotherapy with rituximab to produce a turmour-free graft as well as the safety of retuximab prior to stem cell harvest and post high-dose chemotherapy. Twenty two patients with B-cell non-Hodgkin's lymphoma were enrolled either having aggressive large-cell disease in relapse or at high/high-intermediate risk of relapse, or refractory lymphoma or mantle cell lymphoma, or indolent lymphoma. Chemotherapy consisted of ifosfamide 2 g/m2, days 1-3 with mesna, etoposide 100 mg/m2, days 1-3, and mitoxantrone 8 mg/m2 day 1, with figrastim. Rituximab was given at 375 mg/m2 for 4 doses. An encouraging overall response rate of 90%, including 11 CRs was achieved. CD34+ cells were successfully mobilized in 18 or 19 patients analysed so far with a median number of 3.4 x 10(6) cells/kg. The combination of ifosfamide-based chemotherapy with rituximab significantly reduced the number of contaminating B-cells in the stem cell product and so far there has only been a single relapse post high-dose chemotherapy with autologous haematopoietic transplant. The RIME regimen was generally well tolerated with minimal non-haematological toxicity and most of the treatment was done completely on an outpatient basis. Haematological toxicity was manageable with filgrastim, there were some infectious complications. PMID- 11486405 TI - Ifosfamide-based chemotherapy regimens in treatment of lymphoma: the M.D. Anderson experience. AB - Investigators at M.D. Anderson Cancer Center in Houston first began investigations with the drug ifosfamide in the early 1970s. This alkylating agent has proven to be a valuable drug with easily manageable side effects in therapy of lymphomas. They have described prognostic factors for relapsed lymphomas using this drug in various combinations and have found it of major value in both cytoreduction and stem cell mobilization prior to transplant. The drug has subsequently become part of many established regimens for treatment of lymphoma, and newer combinations containing this drug should be further studied. PMID- 11486406 TI - Backscattering X-ray standing waves in the XUV region. AB - It is demonstrated that Bragg reflection of XUV radiation can be used to study structural properties of crystalline materials with large unit cells. A standing wave field is formed in a layered TiSe2 single crystal for a near-backscattering geometry (theta = 88.5 degrees). The partial electron yield is measured as a function of photon energy across the (001) Bragg reflection condition (hv approximately equal to 1033 eV) and its characteristic modulation is compared with the results derived from dynamical diffraction theory in the two-wave approximation. The data reveal a large amount of disorder along the c-axis. PMID- 11486407 TI - X-ray magnetic circular dichroism imaging with hard X-rays. AB - X-ray polarization-contrast images resulting from X-ray magnetic circular dichroism (XMCD) in the hard X-ray region have been successfully recorded for the first time. The apparatus used consisted of an X-ray polarizer, double X-ray phase retarders, and a high-spatial-resolution X-ray charge-coupled-device detector. The sample used was a hexagonal-close-packed cobalt polycrystal foil having a thickness of about 4 microns. The X-ray polarization-contrast image resulting from XMCD was observed at a photon energy of 10 eV above the cobalt K absorption edge (7709 eV). The observed contrast in the image was reversed by inversion of the magnetic field. Furthermore, the contrast was reversed again at a photon energy of 32 eV above the cobalt K-absorption edge. PMID- 11486408 TI - Synchrotron radiation circular dichroism spectroscopy: vacuum ultraviolet irradiation does not damage protein integrity. AB - Synchrotron radiation circular dichroism (SRCD) spectroscopy is an emerging technique for sensitive determination of protein secondary structures and for monitoring of conformational changes. An important issue for its adoption as a useful technique is whether the high-intensity low-wavelength vacuum ultraviolet radiation in the SRCD chemically damages proteins. In this paper, using horse myoglobin as a test sample, it is shown that extensive irradiation in the SRCD does not produce any change in the chemical nature of the protein as detected by either SDS gel electrophoresis or mass spectrometry. In addition, no changes in the protein secondary structure are detectable from the SRCD spectra after extensive exposure to the SRCD beam. PMID- 11486409 TI - Preliminary experiment of fluorescent X-ray computed tomography to detect dual agents for biological study. AB - The simultaneous observation of various information, such as blood flow, tissue metabolism and distribution of receptors, is quite important in order to understand the functional state of biomedical objects. The simultaneous detectability of contrast agents by fluorescent X-ray computed tomography (FXCT) with synchrotron radiation is examined in this study. The system consisted of a silicon (111) double-crystal monochromator, an X-ray slit system, a scanning table, a PIN diode, a highly purified germanium detector and an X-ray charge coupled device (CCD) camera. The monochromatic X-ray beam energy was adjusted to 37.0 keV and collimated into a pencil beam of 1 x 1 mm. The fluorescent spectra of the K alpha lines for iodine and xenon were detected simultaneously. FXCT could image the distribution of both iodine and xenon agents in a phantom clearly and the contrast ratio was significantly better than that of transmission X-ray computed tomography images. PMID- 11486410 TI - Phase determination by wavelength-modulated diffraction. I. Centrosymmetric case. AB - Wavelength-modulated diffraction was developed by Iwasaki, Yurugi & Yoshimura [Acta Cryst. (1999), A55, 864-870] as a method for phase determination, in which the intensity of Bragg reflections is recorded using radiation whose wavelength is changing continually over a range in the vicinity of the absorption edge of an atom in the crystal. Using a ferrocene derivative crystal (chemical formula C36H32O7Fe, space group P2(1)/a) with the Fe atoms chosen as anomalous scatterers, measurements were made of the intensity gradient dI/d lambda of the reflections with an imaging plate as a detector on a synchrotron radiation source at Ritsumeikan University. In the case of a centrosymmetric crystal, the phase of the structure factor could be derived by measuring only the sign of dI/d lambda at one wavelength in the range. Of 104 reflections measured, the correct phase was assigned to 101 reflections. A discussion is given on the errors involved and on the limits of application of the method. PMID- 11486411 TI - A 2.3 to 25 keV XAS beamline at LNLS. AB - The LNLS XAS beamline has been operating for external users since July 1997. Many facilities and improvements have been progressively added to it, extending the range of applications. Here, a technical description of the main beamline components is given, and results concerning important points, such as available flux at low and high energies, harmonic contamination, energy resolution and stability, are presented. Some key results are given to demonstrate the beamline performance and limitations. It is shown that the beamline can cover a large energy range, starting from the rather low energy of 2.3 keV up to 25 keV. PMID- 11486412 TI - A new ray-tracing program RIGTRACE for X-ray optical systems. AB - A new ray-tracing simulation program, RIGTRACE, has been developed specifically for the X-ray optics range. It traces consecutively each ray, following its path from the source to the observation plane, and treats diffraction by a monochromator crystal by adopting the Darwin-Prins theory so that it may be seen how rays of slightly different energies and incident angles reach the plane. It is also possible to treat the case of the laboratory system, in which the optical elements are disposed close to each other. Examples of the application are provided. PMID- 11486413 TI - A new implementation of the molecular replacement method using a six-dimensional Patterson vector search. AB - The current molecular replacement programs are primarily implemented in reciprocal space. In this paper a new implementation in direct (real) space is proposed by matching the model atomic vectors with the vectors in the Patterson vector space using a six-dimensional exhaustive search method. It is shown that this implementation can find the correct rotations and translations of alpha helices in a myoglobin crystal structure using experimental diffraction data at 2 A resolution. A comparison with previous Patterson vector search methods is discussed. PMID- 11486414 TI - Wet-etched diffractive lenses for hard X-rays. AB - A method for the fabrication of linear transmission Fresnel zone plates for X rays in the 8-15 keV photon energy range is presented. The diffractive elements are generated by electron-beam lithography and chemical wet etching of (110) oriented silicon membrane substrates. Diffractive structures with aspect ratios of more than 30 for 300 nm-wide structures were obtained. The diffraction efficiency of such a lens was measured for 13.3 keV radiation to be 20%. PMID- 11486416 TI - Non-reciprocal X-ray linear dichroism. AB - Recently, a new optical effect was observed by Goulon, Rogalev, Goulon-Ginet, Benayoun, Paolasini, Brouder, Malgrange & Metcalf [Phys. Rev. Lett. (2000), 85, 4385-4388] using linearly polarized synchrotron radiation. The authors have correctly analysed the effect using the mathematical framework of Buckingham and Barron. A brief review is given with the underlying simplicity discussed from a symmetry point of view. PMID- 11486415 TI - Counting the number of disulfides and thiol groups in proteins and a novel approach for determining the local pKa for cysteine groups in proteins in vivo. AB - X-ray Absorption Spectroscopy (XAS) is a powerful tool to investigate sulfur in biological molecules. The spectral features are sensitive to the local electronic and geometric environment of the atom; thus, they constitute a fingerprint of the different chemical forms in which the sulfur is present. This allows straightforward detection of the ratio between free thiols and disulfides. Intra- or inter-molecular disulfide bond formation between residues plays an important role in structural and conformational changes in proteins, and such changes can be investigated using sulfur XAS. Also, a thiolate-disulfide equilibrium is involved in the regulation of the redox potential in the cells by means of modulating the concentrations of the reduced (thiolate) and oxidized (disulfide) form of the tripeptide glutathione. Thus, we can monitor the redox state of a cell by means of sulfur XAS. Thiols also exhibit an acid-base equilibrium, and sulfur XAS can be used to determine the local pKa of the -SH group. Here we report examples of how sulfur XAS has been used for these applications. PMID- 11486417 TI - [Neutropenia in childhood]. AB - The discovery of a neutropenia is a common problem in pediatric population. Some children with neutropenia have an uncomplicated clinical course and require no treatment. However, some patients are more severely affected and present a management challenge for the pediatrician and for the hematologist. The clinical approach to these children is directed primarily to understand the etiology of neutropenia in order to define a correct therapeutic behaviour, such as glucocorticoids, intravenous immunoglobulins or, more recently, rhG-CSF. In our article we have reviewed the etiology, the clinical course and the therapeutic options of constitutional and acquired neutropenias. PMID- 11486418 TI - [Anti-Pseudomonas aeruginosa antibodies, circulating immune complexes, and anticytoplasm antibodies of neutrophils in patients with cystic fibrosis with and without Pseudomonas aeruginosa colonization]. AB - Chronic lung infections, mainly due to Pseudomonas aeruginosa (Pa), account for the most of the morbidity and mortality in CF patients. The pathogenic factors predisposing to airway colonization are still nuclear. Host's immune response is not only poorly protective but can also act as a damaging factor in the development of the disease. Moreover, clinical manifestations of an overactive immune response, including vasculitis and arthropathy, have been recently described in the CF population. To deepen factors involved in the pathogenesis of lung injury, we evaluated the presence of anti-Pa precipitating antibodies, circulating immune complexes (CIC) and antineutrophil cytoplasmic antibodies (ANCA) in sera from 50 CF patients colonized and not by Pa. Number of anti-Pa precipitins was significantly different in chronically and in not colonized patients (p < 0.001; t = 7.75). Anti-Pa antibodies were positively correlated to age (p = 0.002, r = 0.42) and inversely correlated to lung function parameters (p = 0.031 r = -0.35 with respect to FVC). Mean C3-CIC levels in the sera were statistically higher in chronically colonized patients (p = 0.013; t = 2.57); while there was not a significant difference with respect to C1q-CIC values. Four patients, all chronically colonized by Pa, were ANCA-positive at indirect immunofluorescence, showing a cytoplasmic pattern. All 50 patients were found to be negative when tested for anti-mpo and anti-pr3 antibodies with ELISA. In conclusion, our data demonstrate that persistence of Pa provides a stimulus for chronic inflammation and the immune response in CF patients, leading to anti-Pa antibodies and CIC production. Even in the face of further research, we speculate that c-ANCA production is secondary to neutrophil activation with a consequent release of cytoplasmic enzymes that perpetuate and increase the inflammatory process. Since anti-mpo and anti-pr3 antibodies rarely occur in CF patients we suggest that other antigenic specificities may be involved in ANCA production. PMID- 11486419 TI - Fetal vesicoureteral reflux: neonatal findings and follow-up study. AB - To examine features of primary vesicoureteric reflux (VUR) at birth following prenatal hydronephrosis and to describe the course of congenital damage in the first two years of life. We reviewed the records of 382 patients with prenatally diagnosed dilated urinary tracts seen during a 15-years period. When fetal hydronephrosis was confirmed at birth, voiding cystography was performed. If a VUR was diagnosed dimercaptosuccinic acid (DMSA) renography, at 1 month of age, was also performed. Cystography and DMSA renography were repeated and 1 and 2 years of age. Of 382 patients with prenatal hydronephrosis, 68 (17.8%) had primary VUR (50 male and 18 female, M:F = 3:1). VUR was unilateral in 24 patients and bilateral in 44 for a total of 112 refluxing renal units (RRUs). High-grade VUR (grade > or = 4) was found in 70 (62.5%) RRUs, and 60 (85.7%) were boys. Grade V reflux was observed in 96.5% of males. At birth, 22 (20%) RRUs had moderate impairment of renal function. In 12 (10%) RRUs (grade > or = 4) was demonstrated by a severe decrease in renal function. In all cases no focal scarring was noted. Eight infants (all males), with grade 5 degrees VUR, in the first six months of life had surgical treatment. The majority of the RRUs (110/112 = 89%) were followed medically. Mild reflux (grade I-III) had resolved in 81% RRUs, severe reflux (grade > or = 4) had resolved in 38% RRUs. Serial renal scans during the follow-up showed no progression of renal damage. VUR diagnosed at birth on prenatal ultrasonography is associated with congenital damage, with males affected more often than females. In patients without infection, after two years, modification of renal function or renal scars are very rarely observed. PMID- 11486420 TI - [Bone metabolism markers in thalassemia]. AB - The A.A. performed a screening on 113 patients affected by beta-thalassemia major ranging in age between 2 and 40 years, randomized among those which come to the Microcitemic Center of our Institute, and in a control group. In everybody, serum levels of calcium, phosphate, parathyroid hormone (PTH), calcitonin and 25-OH vitamin D were measured. Average serum levels of PTH were significantly (P < 0.001) lower in patients than controls and 12.4% of them were clearly under normal range, especially in the group above 16 years of age. Also serum levels of 25-OH vitamin D were lower in thalassemic subjects than in controls, because of the presence of 32 patients with values under normal limit. Our results are in agreement with current literature that underline the increasing incidence of endocrine complications in thalassemic patients which undergo to high transfusion regimens, owing to the increase of emosiderosis due to the low compliance to iron chelation therapy. Controversial is the pathogenesis of the absence of hypocalcemia in many patients with hypoparathyroidism and the determinism of the deficit of vitamin D. PMID- 11486421 TI - [Approach to constipation in children]. AB - This article presents a diagnostic and therapeutic approach to children with constipation. Causes of constipation can be organic or non-organic (functional). Functional constipation may lead to functional fecal retention. The history and physical examination are most important. Laboratory and radiologic examinations are not warranted in the majority of children. PMID- 11486422 TI - [Sedation in pediatric digestive endoscopy]. AB - Sedation for children doing diagnostic or operative pediatric gastrointestinal endoscopy (PE) procedures is performed differently over the world and no consensus is yet agreed on the best paediatric endoscopy sedation (PES). Some centres do not use any sedation, especially in infants, most centre use some form of sedation: conscious sedation, deep sedation and general anaesthesia. We review sedation drugs and describe our centre protocol on 188 consecutive PE: oral premedication with flunitrazepam (0.05 mg/kg/dose) at least 30 min before procedure, petidine (1 mg/kg) followed by increasing boluses of midazolam (0.05 mg/kg up to a maximal 0.2 mg/kg or 5 mg) were given i.v. to obtain a conscious sedation. All PE could be performed and ended safely, PES resulted satisfactory in approximately 65% of patient having conscious sedation. SaO2 < 90% was observed in 2% of cases, one child had a respiratory depression after PE that resolved with flumanezil. Endoscopy and sedation was always performed by the PE team in the immediate vicinity of anaesthesiologists at work. PE can be safely performed with conscious sedation. Basic and advanced resuscitation skills are needed for the PE team who wish to perform both endoscopic and sedation procedures. PMID- 11486424 TI - [Cardiologic study of 10 patients with Duchenne muscular dystrophy(DMD): personal experience]. AB - Duchenne (DMD) and Becker (BMD) muscular dystrophies are an heterogeneous group of diseases characterized by a progressive muscular degeneration. The locus involved is localized on short arm of chromosome X (2.1) and the gene product has been called dystrophin. The total dystrophin deficiency leads to DMD while the reduction of dystrophin expression to BMD. The dystrophin has an elevated molecular weight (427 Kd) with different isoforms expressed in skeletal muscular system, central nervous system and myocardium tissue. The cardiovascular involvement is up to the muscular dystrophy: in the DMD patients it is secondary to the progressive course and it usually seen in wheel-chair patients. We have retrospectively evaluated through electrophysiology (EKG) and Echo (M-mode and 2 dimensional) studies 10 DMD patients to define the heart involvement. PMID- 11486423 TI - [Inhibin B immunocytochemistry for the prognosis assessment of undescended testis damage in children]. AB - Inhibin B immunocytochemistry of both the alpha- and beta-subunits was studied in testicular biopsies from 18 prepubertal and postpubertal patients, with unilateral or bilateral cryptorchism. The present investigation was carried out to seek any prognostic significance for the expected fertility of such subjects in adulthood. All samples were also evaluated by histological and morphometric assessments, according a 1-6 grading sy-stem. In this way, the individual testicular changes were scored by quantitating tubular and germ cell hypoplasia, Sertoli cell hyperplasia, and peritubular fibrosis, where present. The results showed that in bilateral maldescended testes an unexpected expression often occurred of inhibin B beta-subunit in Sertoli cells, while inhibin B alpha subunit there did not, denoting an early developmental arrest of the testis. It co-related with the high grade testicular damages, as a poor predictor of spermatogenesis. Unlike, unilateral retained testes mainly expressed inhibin B alpha-subunit, irrespective of tubular changes. In the latter instance, different pathogenetic factors of imbalanced testicular regulation can be perspected, other than the Inhibin-Activin system. PMID- 11486425 TI - [Type 1 Arnold Chiari malformation in a subject with Noonan syndrome]. AB - We report a case of Noonan syndrome with characteristic facies, short stature, defect cardiac, epilepsy in a 15 months old girl with Arnold Chiari malformation type 1. We discuss our observations in relation to Noonan syndrome and ACM I. PMID- 11486426 TI - [Severe and unusual clinical manifestations of intolerance to cow's milk protein in 3 patients under 12 months of age]. AB - Cow's Milk Protein Allergy/Intolerance (CMPA/CMPI), due to immunologic hypersensitivity to one or more milk proteins, includes several clinical presentations. We report three unusual cases of CMPA/CMPI, illustrating uncommon and misinterpreted clinical features, with the purpose to provide further evidence of the variety of CMPA/CMPI presentations in infancy. Our study shows that extreme care in assessing the clinical and feeding history of the patient is extremely important for the prompt and correct diagnosis of CMPA/CMPI in infants. PMID- 11486427 TI - [Charge syndrome: long-term survival. Report of a case]. AB - Charge association is a relatively rare pattern of congenital anomalies whose incidence is approximately 1:10.000. We describe our experience on a young boy (Cariotype 46-XY) with associated oesophageal atresia survived up to 12 years. We analyse the most relevant difficulties occurred in the treatment of this patient and we underline the importance of performing an early tracheostomy in order to improve the respiratory impairment. The management of this multiple complex malformations requests a multidisciplinary approach during the life. PMID- 11486428 TI - [Pseudoallergies]. AB - Pseudo-allergic-reactions (PAR) are clinical manifestations including urticaria, angioedema, conjunctivitis, rhinitis, asthma, and anaphylaxis. The prevalence of PAR ranges from 0.1% to 75% according to various studies. The pathogenetic mechanism of these diseases is not immunologically mediated. Food, additives, and drugs are the main responsibilities for PAR. The diagnosis of PAR is characterized by the absence of specific IgE for the suspected products. The absence of immunological mechanisms is confirmed by in vitro and in vivo tests. The treatment of PAR is similar to that of allergic diseases (antihistamine drugs, steroids, B2 agonists, epinephrine). PMID- 11486429 TI - Healthy-living centres. Nipped in the bid. AB - Applicants for healthy living centre funding can find the process very time consuming. Fewer than 100 bids have been approved so far, from more than 1,000 applications. There are concerns about whether the projects will be taken on by primary care trusts once the lottery funding runs out. PMID- 11486430 TI - Public health. How to get promoted. AB - Public health professionals are uncertain about the future of the specialty. Organisational changes will require developing the public health capacity of those outside the profession. Public health professionals will need to develop influencing skills. PMID- 11486431 TI - Primary care. The view from above. AB - The early days of a large PCT found many staff felt ambiguous about the new organisation. It is valuable to define roles and responsibilities early on. PCTs need to take a strategic approach to developing services. PMID- 11486432 TI - First person. From our own correspondents. PMID- 11486433 TI - The effect of laser assisted uvulopalatoplasty on the sense of smell and taste. AB - AIM: To assess the effect, if any, of Laser Assisted Uvulo Palatoplasty (LAUP) on the sense of smell and taste. MATERIALS AND METHODS: A prospective study examined 33 patients undergoing LAUP for snoring. Significant sinonasal pathology was excluded in all patients by a complete examination including rigid nasal endoscopy. None of the cases had any systemic illness or drug history that was likely to affect the sense of smell or taste. Twenty-eight patients were male and five female. The mean age was 44 years (range, 25-67). Twelve patients (36%) were regular smokers, and eight patients (24%) were ex-smokers all of whom reported a change in smell and taste after stopping. Preoperative assessment utilised a visual analogue score of smell and taste perception, the University of Pennsylvania Smell Identification Test (UPSIT) for the assessment of olfaction (maximum score of 40), and whole mouth testing for solutions of NaCl, sucrose, quinine, citric acid and acetic acid. LAUP technique was standardised for each case and reassessment was performed 3 months postoperatively. RESULTS: Three patients failed to complete the study and were excluded. The mean values for visual analogue score pre and post surgery were 9.1 and 8.9 respectively, which was not significant using Wilcoxon Rank test (P = 0.257). The mean value of the UPSIT scores before surgery were 32.3 (+4.0), range, 24-39, and after surgery 31.2 (+4.3), range 20-37. There was no significant difference in pre and post UPSIT scores (p = 0.068). Whole mouth taste testing had a preoperative mean value of 4.1 and a postoperative value of 4.2. The difference was not statistically significant, Wilcoxon Rank test (p = 0.317). CONCLUSION: In this study there was no significant change in the patients' perception of smell and taste or in their objective measurement following a standardised Laser Assisted Uvulopalatoplasty. PMID- 11486434 TI - Breast cancer metastasis presenting as ethmoiditis. AB - Metastasis from primary tumours to the paranasal sinuses is infrequent. We report an unusual case of breast cancer metastasis presenting as ethmoiditis in MRI. MRI changes are unspecific and sometimes inflammatory lesions can not be distinguished from neoplastic lesions. Inflammatory changes in the paranasal sinuses are also frequently noted on MRI even in normal persons without disease. A high index of metastasis suspicion in any patient with breast cancer must be kept in mind. PMID- 11486435 TI - Pneumatization of the concha inferior as a cause of nasal obstruction. AB - Chronic nasal obstruction is a common disorder. Hypertrophy of the inferior turbinates is responsible for nasal obstruction more frequently than it is commonly thought. A pneumatized inferior turbinate has recently been described as a cause for nasal obstruction and only two cases have been reported until now. Inferior nasal turbinate develops by endochondral ossification of components of the mesethmoid and ectethmoid. The chondral framework of the inferior turbinate consists of a double lamella and two separate ossification centers that develop between the fifth and seventh month of fetal life. The separate ossification centers meet by the eighth fetal month. During ossification, the inferior turbinate detaches from the ectethmoid and becomes an independent bony structure. During that time the epithelium may misinvaginate into double lamellas and such double lamellas formed by the inferior turbinate may become persistent. A patient was referred to our clinic with headaches and nasal obstruction. A CT scan was performed which showed that the right lower concha was pneumatized. The headache of the patient disappeared after partial resection of the lower and middle turbinate. PMID- 11486436 TI - Nasopharyngeal epignathus causing nasal airway blocking in a 4-year old child. AB - Congenital oral teratoma or epignathus is a very rare lesion, especially in older children. We report on a 4-year old female infant presenting with nasal blocking, little watery discharge and recurrent otitis. First adenoids were suspected. An endoscopic examination of the nose and nasopharynx demonstrated an irregular mass in the nasopharynx, covered partly by mucosa. A MRI-scan (T1-weighted) showed fat intensity. Histological examination revealed a teratomatous malformation. Although rare, teratoma should be included in the differential diagnosis of a nasopharyngeal mass in the paediatric age group. PMID- 11486437 TI - Leiomyosarcoma of the ethmoidal cells. AB - Leiomyosarcomas of the paranasal sinuses are rare malignant tumors. A case of a 68-year-old female with leiomyosarcoma of the ethmoidal cells is presented. Since half a year she had a stuffed nose on both sides. Preoperatively, several attacks of epistaxis on the right side occurred. CT scans showed a tumor of the ethmoidal cells on the right side. The tumor was completely removed via a functional endoscopic endonasal approach and right sphenoethmoidectomy and maxillary sinus surgery. The operation was followed by a radiotherapy with 72 Gy. Up to 29 months after the operation local recurrence could not be observed. Endonasal tumor resection followed by radiotherapy in a case of leiomyosarcoma without invasion of orbit and skull base can allow tumor control. PMID- 11486438 TI - Inferior concha bullosa--a radiological and clinical rarity. PMID- 11486439 TI - Nasal inflammation and anti-inflammatory treatment. Semantics or clinical reality. AB - In recent years there has been a tremendous development in molecular biology and with that an improved understanding of the immunological and inflammatory background for rhinitis. However, this progress has not yet had any influence on diagnosis or choice of treatment. Today it is emphasized that allergic rhinitis is an inflammatory disease. However, the majority of allergic rhinitis symptoms are caused by histamine, which can be released from a non-inflamed mucous membrane. Thus, the role of inflammation may be overestimated as a cause of rhinitis symptoms. It is often claimed that the 2nd generation antihistamines have non-H1 mediated anti-inflammatory effects of clinical significance. However, the large majority of published clinical data speaks against this hypothesis. Corticosteroids do not, as often believed have a general anti-inflammatory effect in the nose. They are highly effective in a disease associated with eosinophil dominated inflammation (e.g. allergic rhinitis), but not in a disease associated with neutrophil-dominated inflammation (e.g. the common cold). It is recommended that drugs are used merely based on a thorough cost-risk-benefit-patient compliance analysis in the single patient and disease entity with little attention being paid to the assumed mode of action of the drug, which may or may not be of clinical relevance. PMID- 11486440 TI - The role of endonasal surgery in the management of frontoethmoidal osteomas. AB - Recently endonasal surgery has been considered to be a valuable contribution in the management of paranasal sinus osteoma. A retrospective evaluation study of 34 frontoethmoidal osteomas (23 frontal and 11 ethmoidal osteomas) treated at a tertiary care facility from 1990 to 1999 is presented. Twenty three osteomas (68%) were resected endonasally. Eleven osteomas (32%) were removed using an osteoplastic frontal sinus approach with coronal incision. In 5 cases of huge osteomas originating at the anterior frontal sinus wall, reconstruction of the resected anterior-frontal sinus wall was achieved by autologous outer table grafts harvested from the parietal region. Endoscopic and radiological follow-up ranging from 1 to 32 months showed three incomplete endonasal osteoma resections. Complete osteoma removal was achieved via endonasal revision surgery in two of these cases, while the third small residual osteoma remains under observation. There was no case of osteoplastic osteoma removal where incomplete osteoma resection became obvious during follow-up. Ethmoidal osteomas without extrasinusal extension can be resected endonasally. The endonasal approach should be considered also for frontal sinus osteomas if (1) sufficient frontal sinus access can be achieved endonasally, (2) the osteoma is placed medially to a virtual sagittal plane through the lamina papyracea, and (3) the tumour base is at the inferior part of the posterior frontal sinus wall. We favour the osteoplastic frontal sinus approach with coronal incision if an external approach is required to achieve tumour resection with the best aesthetic results. PMID- 11486441 TI - Endoscopic optic nerve decompression for the treatment of traumatic optic neuropathy. AB - Optic nerve decompression has been accomplished by a variety of procedures. Since 1995, endoscopic optic nerve decompression (EOND) has been used to treat traumatic optic neuropathy (TON) patients in our hospital after medical treatment failed. To date, 17 TON patients have received EOND in our hospital and have been followed up for more than half a year. After decompression, vision improved in 9 patients, remained the same in 6, and became worse in 2. The dura was incidentally exposed during the operation in 1 patient. We conclude that it is easier and more precise to perform optic nerve decompression by EOND than by other optic nerve decompression procedures. However, its efficacy still needs further investigation. PMID- 11486442 TI - Topical corticosteroids in chronic rhinosinusitis: a randomized, double-blind, placebo-controlled trial using fluticasone propionate aqueous nasal spray. AB - Chronic rhinosinusitis (CRS) is a recalcitrant inflammatory process which has a marked detrimental impact on quality of life. At the present there is no cure for this condition, measures are taken to stop progression, and provide symptomatic relief. Topical corticosteroids are commonly prescribed in the management of CRS, but few trials show effectiveness in clinical settings. We set up a randomized, double-blind, placebo-controlled trial to study the effectiveness of a topical corticosteroid agent--fluticasone propionate aqueous nasal spray (FPANS) in patients with CRS. We measured symptoms, diary card, and rigid endoscopy scores, acoustic rhinometry, middle meatal swabs, blood tests--CRP, ESR, WBC, and eosinophil count. Measurements were done at the start of the trial, at 8 weeks, and 16 weeks where possible. Twenty-two patients completed the trial, 9 received FPANS, and 13 had placebo. There was no difference between the 2 groups on all counts. When patients were considered as one group, there was an improvement in the diary card scores (p = 0.054), comparing baseline to 8 or 16 weeks. There was no evidence that the regular use of topical corticosteroid increased the risk of developing an infection. An important observation was that the topical corticosteroid did not precipitate acute sinusitis. There is compelling evidence that topical corticosteroids down-regulate cytokine expression, and it is likely that a larger, and longer multi-centre trial may prove their efficacy in CRS. PMID- 11486443 TI - Effect of nasal dilators on pleasantness, intensity and sampling behaviors of foods in the oral cavity. AB - The present study assessed the effect of nasal dilators on ratings of food intensity and pleasantness. Participants wearing the dilators rated foods in the oral cavity as less pleasant and more intense than did those participants wearing a placebo strip. In addition, they consumed less of the test stimuli. Significant interactions were noted between food stimuli and placebo vs. nasal dilator strip conditions, indicating certain food qualities, particularly initial pleasantness, combine to produce enhanced changes in perception. Changes in the intensity and pleasantness of foods was most pronounced in foods characterized as initially pleasant. A review of studies to date indicate that both foods and odors are perceived as more intense and less pleasant during nasal dilation, suggesting an impact of the dilators on both retronasal and orthonasal air flow. PMID- 11486444 TI - Effect of nasal dilators on nasal structures, sniffing strategies, and olfactory ability. AB - This paper describes the effects that nasal dilators have on olfactory ability. Experimental results demonstrate that nasal dilators increase odorant identification, lower odorant threshold, and increase perceptual odorant intensity. In other experiments, magnetic resonance imaging (MRI) data demonstrates that the size of the nasal cavity especially around the region of the nasal valve is increased when nasal dilators are worn. Additionally, pneumotachograph data demonstrates that during a sniff, the peak flow, maximum flow rate, volume, and duration are all increased when nasal dilators are worn. Taken together, the increase in olfactory ability can most easily be explained by an increase in both the amount and the proportion of inspired odorant molecules that are directed to the olfactory mucosa and are, therefore, available for odorant perception. PMID- 11486445 TI - Basic fibroblast growth factor in nasal polyps immunohistochemical and quantitative findings. AB - Basic fibroblast growth factor (bFGF) is a mediator with potent mitogenic properties. Increased amounts of this mediator have been demonstrated in damaged lung tissue, and it has been suggested to increase the healing of gastro-duodenal ulcers. In order to quantify the amounts and document the localization of bFGF in nasal polyps, polyp tissue from 12 patients undergoing polypectomy was analyzed by ELISA and immunohistochemistry. Mucosa from the inferior turbinate was analyzed in the same manner for comparison. The amount of bFGF detected in polyp tissue was significantly higher than that in turbinate mucosa. The amount of bFGF was also significantly higher in the group of patients with high degree of inflammation. The immunohistochemical findings demonstrated abundant bFGF activity mainly in the glandular acini, in the epithelium, in infiltrating inflammatory cells and in the vessel walls. We propose that bFGF may contribute in a significant way to the formation of nasal polyps. PMID- 11486446 TI - Evaluation of the importance of head and probe stabilisation in acoustic rhinometry. AB - As yet there is no established procedure to ensure the repeatability of acoustic rhinometry measurements although anecdotal evidence suggests that instrument fixation improves repeatability. The aim of this study is to validate the methodology of acoustic rhinometry and determine whether instrument fixation and head stabilisation is necessary. Four methods were compared in fifteen healthy volunteers, after nasal decongestion: A) Patient holding the probe (patient held), B) Probe fixed in a probe stand (probe-stand), C) Probe fixed in stand and head stabilised in head rest (head-rest), D) Examiner holding the probe (examiner performed). The two minimum cross-sectional areas and volume between 0 and 5 cm were recorded. The examiner-performed and probe-stand methods were consistently less variable than the other methods. With examiner-performed method, this was significant (p < 0.05) versus head-rest and patient-held methods for both measures of minimum cross-sectional area. For nasal volume the examiner-performed method was significantly (p < 0.05) less variable than the head-rest method. In conclusion, examiner-performed acoustic rhinometry is more repeatable than combined head stabilisation and instrument fixation and therefore the use of a head-rest may be unnecessary. Instrument fixation or examiner performed test is also preferable to allowing the patient to position the probe. The repeatability of the probe-stand method was similar to the examiner-performed method. PMID- 11486447 TI - Herpesvirus type 1-8 in sinus aspirates from HIV-infected patients and immunocompetent individuals. AB - Sinusitis is frequently occurring in HIV-infected patients, but in a substantial number of cases the etiology is unknown. The purpose of this study was by PCR 1) to determine the prevalence of the eight human herpesviruses in sinus aspirates from 24 HIV-positive/AIDS patients with sinusitis 2) to relate the presence of herpesvirus DNA to clinical and immunological parameters and 3) to compare the prevalence of herpesvirus DNA in sinus aspirates from HIV-infected patients with the prevalence observed in 50 immunocompetent patients with sinusitis. DNA from HSV-1, EBV, CMV and HHV-8 was detected in 8 (33%) of the sinus aspirates from HIV infected patients. In the immunocompetent patients, one of the herpesviruses, HHV 6, was found in one sinus aspirate. These data indicate that herpesviruses are frequently found in sinus aspirates from HIV-infected patients with sinusitis, whereas they do not seem to be related to clinical signs of sinusitis in immunocompetent individuals. The cause of these discrepancies may be due to uncontrolled reactivation of herpesviruses, which is known to occur in immunocompromised individuals. It remains to be established whether the herpesviruses play a pathogenic role in the development of sinusitis in HIV infected patients. PMID- 11486448 TI - Fluconazole vs. amphotericin B for the management of candidaemia in adults: a meta-analysis. AB - The incidence of bloodstream infections caused by Candida species is rising. Few published studies have compared the efficacy of fluconazole with that of amphotericin B. We performed a meta-analysis of the prospective studies that compared fluconazole and amphotericin B for the treatment of candidaemia in adults. Data on total mortality, candidaemia-attributable mortality, efficacy, microbiological failure, and toxicity were extracted from eligible studies. All studies appeared homogeneous with respect to the outcome measures. Most patients were at relatively low risk for death as evidenced by the low average physiologic score and the lack of intense immunosuppression. The odds ratios (OR) of treatment with amphotericin B versus fluconazole and 95% confidence intervals (CI) were as follows: total mortality (OR, 1.06; CI, 0.89-1.25), candidaemia attributable mortality (OR, 1.0; CI, 0.70-1.45), clinical response (OR, 1.14; CI, 0.93-1.39) and microbiological failure according to all Candida species (OR, 0.99; CI, 0.78-1.26). A trend favouring amphotericin B was seen in mycological eradication of non-albicans Candida species (OR, 0.70; CI, 0.47-1.06). Finally, amphotericin B was more toxic than fluconazole (OR, 2.94; CI, 2.14-4.4). In conclusion, fluconazole is as efficacious and less toxic than amphotericin B in stable, not severely immunosuppressed candidaemic patients at low risk for death. However, fluconazole may be less effective than amphotericin B in candidaemias caused by some non-albicans Candida species. PMID- 11486449 TI - First isolation of Cryptococcus neoformans var. gattii from a native jungle tree in the Brazilian Amazon rainforest. AB - Cryptococcus neoformans var. gattii was isolated for the first time from decaying wood in a hollow of a native jungle tree Guettarda acreana, in a wild area of an Amazon rainforest island, in Brazil. The presence of this variety in a virgin environment without either anthropic action or introduced vegetation is discussed with regard to the common knowledge of Cr. neoformans ecology. PMID- 11486450 TI - Sinocranial aspergillosis: a form of central nervous system aspergillosis in south India. AB - Of the 21 patients with aspergillosis of central nervous system seen during the years 1990-1997, 16 (76%) had aspergillosis of sino-cranial origin. The occupation in patients with sino-cranial aspergillosis was either agricultural or manual work and predisposing risk factors were present in only two (12.5%) patients. Skull-base syndromes were the presenting features in 13 patients and three patients presented with features of intracranial space-occupying lesion. Paranasal sinus mass lesions were seen in all the 16 patients. Computerized tomography showed intracranial extradural-enhancing mass lesions in the anterior, middle or posterior cranial fossa in nine (68%) patients, intracranial and orbital lesions in four and orbital lesions in three. Well-formed granuloma with dense fibrosis was the histological feature. Survival rates were not good even after surgical and antifungal chemotherapy. Surgical treatment was subradical in our series. The majority of cases of sinocranial aspergillosis are reported from countries with temperate climates and the high incidence in these regions is probably related to constant exposure to the high spore content of pathogenic Aspergillus species in the 'mouldy' work environment. PMID- 11486451 TI - Yeast infection of burns. AB - In a prospective study, 132 patients were investigated for yeast infection of burn wounds. Ten patients (7.6%) were infected with Candida species. All patients with yeast infections were also infected with bacteria with the exception of one patient who was infected with Candida tropicalis alone. The predominant yeast recovered was Candida krusei. Yeast infection was found to be more common in the younger age group. The isolation of a Candida species alone from one patient and Candida isolation from patients with sepsis in burn wounds indicate a significant role for yeasts in the production of infection in burn wounds. Therefore, special cultures for yeasts are recommended for all cases of burn wound infection. PMID- 11486452 TI - Faecal fungal flora in healthy volunteers and inpatients. AB - The faecal fungal flora was analysed in healthy volunteers and inpatients. Self obtained stool swabs from volunteers (n = 228) and inpatient stool-samples (n = 34) were cultured on Inhibitory-Mould-Agar plates. All yeast isolates were identified. Fungi were detected in 51.8% of volunteers; the majority (88.1%) had single species. The prevalence increased steadily with age. Candida albicans was detected in 62.7%, non-albicans Candida species in 22.0%, yeasts--other than Candida in 20.3% and moulds in 8.5% of volunteers with fungi. No gender-related differences were noted in the prevalence or types of yeast. Candida glabrata and C. krusei were detected in adults only. Intra-household species-similarity (excluding C. albicans) was noted in seven of 31 (22.6%) households with fungi in two or more members. Inpatients had higher prevalence of yeast (88.2%) with a single species in the majority (73.3%). Yeasts other than Candida were less common in inpatients (3.3%; P = 0.013) whereas C. glabrata was significantly more prevalent (33.3 versus 2.5%; P < 0.001). This study delineates the faecal fungal flora in volunteers and inpatients. Most subjects harbour a single species that may be shared with other households. The prevalence is somewhat higher in adults and the types of yeast may vary with age. Finally, C. glabrata appears to be acquired nosocomially. PMID- 11486453 TI - Identification and subtyping of Trichophyton mentagrophytes by random amplified polymorphic DNA. AB - Trichophyton mentagrophytes is one of the most common pathogens for human and animal dermatophytoses and known as a complex species with variable morphology, and ecologic and genetic backgrounds. We performed random amplified polymorphic DNA (RAPD) analysis on 43 human and 18 animal isolates of T. mentagrophytes along with other 10 anamorphic species of dermatophytes and three teleomorphic species of T. mentagrophytes. Using RAPD analysis with primer 5'-ATGGATC(G,C)(G,C)C-3' (ATGS), all T. mentagrophytes strains produced identical band patterns with those of Arthroderma vanbreuseghemii, one of the teleomorphs of T. mentagrophytes, regardless of their phenotypes. Therefore, T. mentagrophytes could be identified by RAPD analysis with primer ATGS. Using RAPD analysis with primer 5'-ATGGATCGGC 3' (ATG) on T. mentagrophytes, human isolates yielded two distinct subgroups related by their colony morphologies at the time of primary isolation from patients. Three morphologic types--cottony, powdery and persicolor--revealed identical bands whereas the granular type lacked one minor band (0.74 kbp). Animal isolates of T. mentagrophytes produced five band patterns and some of them were identical with those of human isolates. With primer 5'-GAAGGCTCCC-3' (OPAO 15), animal isolates of T. mentagrophytes showed diverse band patterns in contrast to the uniform band pattern of human isolates. These results suggest that RAPD analysis may be a useful tool to identify and subtype T. mentagrophytes complex. PMID- 11486454 TI - The definition of Trichophyton rubrum syndrome. AB - Although chronic widespread dermatophyte infection is reported widely in the literature, neither a uniform nomenclature, nor even a clear definition of this syndrome have been established so far. Thus, we suggest Trichophyton rubrum syndrome (TRS) for denomination and define the following obligatory clinical and mycological criteria for TRS. (A) Skin lesions at the following four sites: (1) feet, often involving soles; (2) hands, often involving palms; (3) nails; and (4) at least one lesion in another location than (1) (2) or (3), except for groins. (B) Positive microscopic analyses of potassium hydroxide preparations of skin scrapings in all four locations. (C) Identification of Trichophyton rubrum by cell culture at three of the four locations at least. For diagnosis of TRS the criteria (A) and (B) and (C) have to be fulfilled. This standardization is a prerequisite for further investigations of underlying mechanisms of this disease. The typical clinical pattern of TRS is illustrated by the presentation of two paradigmatic cases. PMID- 11486455 TI - A double-blind randomized comparative trial: eberconazole 1% cream versus clotrimazole 1% cream twice daily in Candida and dermatophyte skin infections. AB - In a double-blind phase III study the efficacy and tolerance of eberconazole 1% cream was compared with clotrimazole 1% cream, applied twice daily for four consecutive weeks in 157 patients, with mycologically proven cutaneous candidosis (24 patients, 25 infected sites) and in 133 patients with dermatophyte skin infections (141 treated sites). The analysis was carried out by number of infected and treated sites. The four groups of sites and the distribution of target lesions were similar. The distribution of the sum of clinical scores and infecting organisms were also similar. There were no differences between the groups in terms of the range and mean duration of infection. At the end of therapy and on follow-up in skin candidosis the proportion of patients with effective treatment was 73 and 50% for clotrimazole and eberconazole, respectively. Premature termination due to adverse effects was 26.7% for clotrimazole and 20% for eberconazole. At the end of treatment of dermatophyte infections the proportion of patients with effective treatment was 46% for clotrimazole and 61% for eberconazole. At the overall assessment (6 weeks after the end of treatment) eberconazole was judged effective in 72% of treated sites, whereas clotrimazole was effective in 61%, this difference was not statistically significant (P = 0.15). There were three relapses (4%) in the clotrimazole treated group and one relapse (1%) in the eberconazole group. One patient treated with eberconazole cream (1%) had a premature termination due to adverse events attributable to the medication. PMID- 11486456 TI - Usefulness of lanoconazole (Astat) cream in the treatment of hyperkeratotic type tinea pedis. Comparative study of monotherapy and combination therapy with 10% urea ointment (Pastaron). AB - Hyperkeratotic type tinea pedis is a refractory type of superficial dermatomycosis. Treatment for hyperkeratotic type tinea pedis is mainly with oral antimycotics, such as griseofulvin, and healing is generally considered to be difficult with only topical antimycotics. In this randomized comparative study, the usefulness of a topical application of 1% lanoconazole cream (Astat) monotherapy (group I) was compared with that of combination therapy with 1% lanoconazole cream and 10% urea ointment (Pastaron) (group II) in a series of patients with hyperkeratotic type tinea pedis. The clinical improvement rates (percentage of 'marked improvement' plus 'moderate improvement') was 70.0% in group I and 95.7% in group II. The fungal eradication rate was 5.0% in group I and 43.5% in group II after 4 weeks of treatment, and was 70.0% and 95.7% after 12 weeks of treatment, respectively. The usefulness rate (percentage of 'very useful' plus 'useful') was 70% in group I and 95.7% in group II. Both lanoconazole monotherapy and the combination therapy with 10% urea ointment were highly effective and safe. Both treatments should be recommended for patients with hyperkeratotic type tinea pedis for whom an oral treatment is not appropriate or for whom a sufficient improvement with oral medications cannot be expected. PMID- 11486457 TI - Age as limiting factor of the efficacy of itraconazole for treatment of onychomycosis. AB - A clinical study was carried out involving 24 patients between 18 and 64 years of age with disto-lateral onychomycosis of finger and/or toenail in whom intermittent therapy with itraconazole was orally administered for 4 months. A follow-up period of 9 months was instituted to monitor the efficacy of the treatment. Clinical and mycological outcome at the end of the study was correlated with a checklist that included age and sex of the patient, site of infection and species of the causative agent. Nails that were free of disease in both gross and mycological examinations were achieved in 58% of our patients. Age was shown to be the only parameter, among those taken into consideration, that was correlated with the cure rate. The association was found to be statistically significant. PMID- 11486458 TI - Case report. Successful outcome of invasive nasal sinus zygomycosis in a child with relapsed acute lymphoblastic leukaemia due to liposomal amphotericin B. AB - We report a case of life-threatening nasal sinus zygomycosis that developed during remission induction therapy for a relapsed acute lymphoblastic leukaemia. The patient was successfully treated with liposomal amphotericin B and granulocyte-colony stimulating factor followed by surgical reconstruction of the resultant cutaneous defect. PMID- 11486459 TI - Case reports. Secondary prophylaxis with liposomal amphotericin B after invasive aspergillosis following treatment for haematological malignancy. AB - We report our recent experience with two cases of invasive pulmonary aspergillosis in patients who were both undergoing chemotherapy, one for acute myeloid leukaemia and the other for primary amyloidosis. Both patients had bad prognostic factors and were in very poor clinical condition, but both recovered from infection after a prolonged therapy with liposomal amphotericin B (AmBisome) without signs of toxicity. PMID- 11486460 TI - Eligibility criteria for Nd-YAG laser treatment of highly symptomatic vitreous floaters. AB - Ten eyes of nine patients were treated for very disturbing vitreous floaters with the technique of Nd-YAG laser vitreolysis. The Scanning Laser Ophthalmoscope (SLO) was used to objectivate the position, the size and the motility of the vitreous floaters with respect to the patient's visual axis, which can be precisely located with the SLO. With this technique it was possible to define more precisely some eligibility criteria for Nd-YAG laser treatment of vitreous floaters and to classify the vitreous floaters in ill-suspended and well suspended floaters in the vitreous body, the well-suspended floaters responding better to treatment compared to the ill-suspended vitreous floaters. The treatment was performed using the Q-Switched Nd-YAG Laser type Nanolas 15S of Alcon. PMID- 11486461 TI - Non penetrating deep sclerectomy (NPDS) with SKGEL implant and/or 5-fluorouracile (5-FU). AB - PURPOSE: To assess effectiveness and safety of DS with reticulated hyaluronic acid implant (SKGEL) and/or 5-Fluorouracile (5-FU). MATERIAL-METHODS: 61 eyes (48 patients) (mean age: 64.5 +/- 10.5 years) with medically uncontrolled open angle glaucoma and comparable surgical risk failure were included in this retrospective non randomized study and were categorized into DS + SKGEL (10 eyes), DS + 5-FU (25 eyes) and DS + SKGEL + 5-FU simultaneously (26 eyes). The 3 groups were comparable in respect of age of patients, diagnosis, severity of VF defects, and bleb failure risk factors. All procedures were performed according to the Kozlov's and Mermoud's technique. Complete ocular examination was carried out preoperatively and at day 1, 7, at 1, 2 and 3 months and every three months thereafter. RESULTS: Mean follow up was 11 +/- 4.9 months and was significantly shorter in group C. For all 61 eyes, mean IOP was significantly decreased from 27.8 +/- 8.6 mm Hg to 15.1 +/- 3.5 mm Hg. Complete (target IOP reached without medication or YAG laser goniopuncture) and qualified (target IOP reached with medication and/or YAG laser goniopuncture) final success rates were respectively of 54% and 90%. Complete success probability was 95% at 6 months and 72% at 12 months. Qualified success probability increased to 89.5% at 12 months. The IOP results, the distribution of success rates and complications were similar within the first postoperative year whatever using SKGEL or intra-operative 5-FU application or both. The need for additional glaucoma medication and the percentage of YAG goniopuncture appeared to be lower in the SKGEL group. Success probability appeared to be improved in SKGEL group comparatively with the other 2 groups. CONCLUSIONS: NPDS is a valid, effective and relatively safe alternative to trabeculectomy although adjunctive medications and/or Nd:YAG goniopuncture are frequently needed. Considering the limitations of our study, we can conclude that the IOP results, success rates and complications are similar within the 1st postoperative year whatever using peroperative 5-FU or SKGEL implant. A longer follow up is needed to confirm that SKGEL implant seems to improve the IOP control and the long term patency of the sclerectomy site by comparison with intra-operative application of 5-FU. PMID- 11486462 TI - [Comparison of preoperative target ametropia and postoperative refraction for three types of lenses and intra-ocular differences (one pliable acrylic, one pliable PMMA-copolymer and one non-pliable PMMA) ]. AB - In this retrospective study we compared the preoperative target ametropia, calculated with the SRK-T formula, with the postoperative refraction after extracapsular extraction by phacoemulsification and implantation of different posterior chamber lenses. The results show, for all 3 types of lenses, a postoperative refraction stability after the first month. According to our criteria, after 6 month, we have respectively for the non foldable PMMA lens, the PMMA-copolymere foldable and the acrylic foldable lens: 78%, 72% and 78% of good results; 13%, 28% and 9% of mild results and 9%, 0% and 13% of bad results. According to our criteria, the acrylic foldable lens has a tendency to move slightly backwards and the foldable PMMA-copolymere lens to move slightly frontwards. The non foldable lens gives pretty good results, however it is difficult to compare this lens with the 2 other lenses because of the large size of the incision. PMID- 11486463 TI - The array multifocal intraocular lens: a new horizon. AB - A retrospective review was conducted of 50 patients with bilateral Array multifocal intraocular lens implantation. The purpose was to assess their visual performance and adaptability to the visual world of multifocality. On binocular visual acuity examination all patients obtained, without any correction, Snellen 20/25 or more for distance and Snellen 20/40 (distant equivalent) or more for near. Contrast sensitivity for far and near was slightly reduced. Occasionally, 44% of the patients use spectacles especially for small print. All patients were very pleased with their functional vision without spectacles and in general had only minor problems with optical aberrations. Factors contributing to success and patient satisfaction are refractive cataract surgery and careful selection of patients through meticulous pre- and postoperative counselling on the new visual status. PMID- 11486464 TI - [Integration of visually impaired children into normal social structures]. AB - The author explains how works a centre for functional rehabilitation and the role of the ophthalmologist. She gives the reasons for an early care of visually impaired children, the aims of the visual stimulations and the conditions for the integration in a normal school. PMID- 11486465 TI - Reconstruction of orbital floor blow-out fractures with silicone implant. AB - AIM: To evaluate the outcome of orbital floor blow-out fractures treated surgically with silicone implant. METHODS: The patients were examined in a prospective study from October 1993 to December 1999. Over this period 11 patients were diagnosed as having orbital floor blow-out fractures. The study was restricted to those who were both treated surgically and followed-up at least three months after the discharge from hospital. Only six patients (five males and one female), ranging in age from 6 to 30 years, fulfilled the inclusion criteria. The fractures occurred during brawls in 3 patients, car accidents in 2 patients and child's play in 1 patient. The diagnosis was based on history, clinical grounds and coronal computed tomography. The surgical procedure was the same in all patients, using silicone implant to reconstruct the defects. The mean follow up time after surgery was 4.8 months. RESULTS: The interval between injury and diagnosis ranged from 6 to 85 days (mean: 34.8 days). Limitation of vertical eye movements was present in all 6 patients, diplopia in 4 patients, enophthalmos in 3 patients and hypoesthesia in the distribution of the infraorbital nerve in 1 patient. The mean interval time between injury and surgery was 55.3 days and the majority of the patients (83%) had late repairs (> 14 days) after injury. Postoperatively, satisfactory results were obtained with regard to limitation of vertical eye movements, diplopia, enophthalmos and hypoesthesia. Only one patient had a persistent and partially reduced enophthalmos. Silicone implant was well tolerated in all 6 cases since complications such as infection, tissue reaction and extrusion were not observed. CONCLUSION: Satisfactory results may be obtained after late repair of orbital floor blow-out fractures. Silicone implant has the potential to be used successfully in orbital floor fractures. PMID- 11486466 TI - [Grave complications of local anesthesia for cataract extraction: reports of 8 cases]. AB - Cataract extraction performed under local anesthesia is not so safe. During a 2 years period, 8 patients have presented a complication: 3 patients are presented with diplopia, 4 patients are presented with eyeball perforation and 1 patient developed a central complication. All those complications are caused by the superior peribulbar injections of anesthetics. A survey of the different etiologies of the superior rectus muscle blow and of eyeball perforation is given. PMID- 11486467 TI - [Ocular Behcet's disease: procedural aspects]. AB - The diagnosis of ocular Behcet is clinical. This affection is characterised by a bilateral anterior and/or posterior recurrent non granulomatous intraocular inflammation. The treatment consists in the use of one or several associated immunomodulators and immunosuppressors. In order to decrease the drug toxicity we prefer to increase the number of associated drugs than to increase the doses in severe resistant cases. The prognosis can be significantly improved by a very strict control of chronic inflammation and of each exacerbation. Interferon alpha could be a good new treatment in the future. PMID- 11486468 TI - Immunology of trachomatous conjunctivitis. AB - Trachoma, a chronic follicular conjunctivitis caused by infection with Chlamydia trachomatis, is a leading cause of preventable blindness. The blinding complications are associated with progressive conjunctival scarring. Our immunohistochemical studies of conjunctival biopsies from children with active trachoma demonstrated the presence of both humoral and cell-mediated immune responses. Antichlamydial antibodies can neutralize Chlamydiae, block attachment and internalization of the organism, and can produce partial immunity. Our observations suggest a role for T-lymphocytes and cell mediated immunity in the genesis of conjunctival scarring. Conjunctival epithelial cells expressed major histocompatibility complex (MHC) class II antigens which might allow conjunctival epithelial cells to present Chlamydial antigens to T-cells enhancing the immune response. The epithelial cells expressing MHC class II antigens might present autoantigens to T-cells leading to induction of an autoimmune reaction. We have demonstrated that the conjunctival epithelial cells from patients with trachoma expressed interleukin (IL)-1 alpha and IL-1 beta. In addition, we have detected cytoplasmic expression of IL-1 alpha, IL-1 beta, tumor necrosis factor alpha and platelet-derived growth factor by macrophages. These cytokines have the potential to influence the remodeling and fibrosis observed in trachoma. Alterations of extracellular matrix components and collagen metabolism occur in the conjunctival tissue from patients with trachoma. New collagen type V formation was noted in active trachoma and scarred trachoma. The conjunctival tissue from patients with active trachoma contained increased amounts of collagen types I, III and IV. Scarred trachoma is characterized by marked increase in basement membrane- collagen IV and marked decrease in collagen types I and III. In addition, we demonstrated increased activity of gelatinase B and numbers of inflammatory cells containing gelatinase B in trachoma patients suggesting that this enzyme might be involved in matrix degradation and promotion of conjunctival scarring in trachoma. PMID- 11486469 TI - Intraocular pressure elevation in a child due to the use of inhalation steroids- a case report. AB - Inhalation steroid therapy can cause ocular hypertension or open angle glaucoma. The authors describe the case of a young girl who presented with raised intraocular pressure and headaches due to the prolonged administration of nasal and inhalation steroids. The ophthalmologist should monitor the intraocular pressure in patients who use inhalation or nasal steroid therapy on a regular base. The physician or paediatrician should be aware of this complication in children with headaches or diminished visual acuity. PMID- 11486470 TI - Gastro-intestinal stromal tumours (GISTs): a six year Zimbabwean experience. Cellular differentiation with immunocytochemistry. AB - OBJECTIVES: To determine the prevalence of GISTs in general, the terminology in use, biological behaviour in terms of benign, borderline and malignant variants and assessment of the different lines of differentiation using immunochemical techniques. DESIGN: A retrospective study. SETTING: Department of Histopathology, University of Zimbabwe Medical School at Parirenyatwa and Harare Central teaching Hospitals. SUBJECTS: Patients with a surgical pathology diagnosis of gastro intestinal related stromal tumours including leiomyosarcomas, leiomyomas and GISTs over a six year period. MAIN OUTCOME MEASURE: Evaluation for different lines of differentiation and malignant potential using immunocytochemical techniques. RESULTS: A total of 24 cases with a primary diagnosis of GISTs over a period of six years were retrieved from the records and of these 22 had material available for evaluation. Four of these were reported as pure GISTs, two of them benign and the other two malignant. Seven cases were reported as leiomyosarcomas. Six of these had evidence of neural differentiation on immunocytochemistry and therefore were strictly not leiomyosarcomas. Of the 11 cases reported as leiomyomas, four (36.4%) had neural differentiation on immunohistochemistry implying a low malignant potential. CONCLUSION: GISTs are not uncommon in this patient population considering that 24 cases were reported over a six year period in a department that averages 11,000 surgical pathology cases per year. Minimal use of immunostains, in addition to general morphological features, can be utilized to determine the line of cellular differentiation and malignant potential of the seemingly benign tumours. PMID- 11486471 TI - Urinary diversion in children with mainly exstrophy and epispadias: alternative to primary bladder closure. AB - OBJECTIVE: To review the long term outcome of children who underwent urinary diversion for various non-malignant conditions and to assess the suitability of urinary diversion as an alternative treatment to primary bladder closure in cases of bladder exstrophy epispadias complex. DESIGN: A retrospective study. SETTING: Institute of Urology, Moshi, Tanzania. SUBJECTS: Fifteen children who underwent urinary diversion for various non-malignant conditions between 1985 and 1997. RESULTS: Fifteen children underwent urinary diversion for: exstrophy epispadias complex (n = 12) neurogenic bladder (n = 2) and trauma (n = 1). Seven children underwent Mainz pouch II diversion, six had classical ureterosigmoidostomy and two had appendicovesicostomy. One patient with a solitary kidney developed ureteral stenosis at the implantation site and was undiverted. Metabolic acidosis was well compensated with none of the patients requiring sodium bicarbonate supplements. All the patients except one were continent. CONCLUSION: We conclude that continent urinary diversion in children offers a viable alternative method for children with bladder exstrophy epispadias complex. PMID- 11486472 TI - Gangrenous bowel in Nigerians. AB - OBJECTIVE: This study is to critically examine gangrenous bowel as seen in a developing country to identify factors associated with mortality and to reappraise the problems in our environment. We also looked at mortality associated factors not considered by other studies from the subregion. DESIGN: We retrospectively reviewed the records of all patients who had gangrenous bowel over a 10 year period (1986 to 1995) at the University of Ilorin Teaching Hospital, Ilorin, Nigeria. SETTING: University of Ilorin Teaching Hospital, Ilorin, Nigeria. SUBJECTS: All patients with gangrenous bowel seen at the University of Ilorin Teaching Hospital, Ilorin Nigeria from January 1986 to December 1995 were included is this study. RESULTS: Ninety five patients were seen with a male to female ratio of 2:1. A high prevalence at age zero to five (17.9%) was noted due to childhood intussusception. The average duration of symptoms before surgery was 5.6 days. External hernia were responsible for 42.1%, intussusception for 22.1% and adhesions for 12.9% of cases. Other rarer and unusual causes of gangrene are also presented. Ileal gangrene was seen in 80.9% of cases. The overall mortality was 35.8% with no significant age and sex differences. Mortality varied directly with the length of gangrenous bowel irrespective of the cause and with the delay in operative intervention up to eight days. Peritonitis and involvement of the colon were also associated with high mortality. CONCLUSION: As a result of late presentation allowing for high diagnostic accuracy, elaborate investigations to determine bowel viability have little role in our setting at present. Improvement in socio-economic status, health education, elective repair of hernia and early operative intervention are recommended to bring down the mortality from gangrenous bowel. PMID- 11486473 TI - Self reported sexual and reproductive behaviour of male and female student nurses and midwives in Bulawayo. AB - OBJECTIVE: To find out more about sexual and reproductive behaviour in male and female student nurses and midwives in Bulawayo. DESIGN: Anonymous self administered questionnaire. SETTING: Schools of Nursing Mpilo and Bulawayo Central Hospitals. SUBJECTS: Three hundred and twenty four students, student nurses (n = 99) and student midwives (n = 225). MAIN OUTCOME MEASURES: Preferred family size, enjoyment and frequency of sexual behaviour, induced abortion, masturbation, pre- and extramarital sex and HIV risk. RESULTS: Male student nurses have far more pre- and extramarital sexual contacts than their female counterparts. Of the married females 67% had only ever had one sexual partner, (this figure was 15% in married males) and 33%, suspect or know that their husband has another sexual partner sometimes. CONCLUSION: Promiscuity is rare in female student nurses. Their risk factors, if any, related to HIV infection are mainly caused by their partners' pre- and extramarital behaviour. HIV testing of these partners is their only realistic hope of reducing their risk. The enjoyment of heterosexual activity is like that in Western Countries although it starts at a later age. PMID- 11486474 TI - Results of intra-operative 0.5mg/ml mitomycin C with 20mg depo steroid in the treatment of primary pterygium. AB - OBJECTIVE: The effectiveness of intra-operative mitomycin C on the recurrence rate of pterygium in patients undergoing primary pterygium excision was assessed. METHOD: 51 consecutive primary pterygia of 46 patients were excised between April and December 1998. 0.5 mg/ml mitomycin C was applied on bare sclera for one minute and 20 mg depot steroid injected subconjunctivally. Post operatively patients were placed on Gutt Dexamethasone 0.1% with antazoline and chloramphenicol 1.0%. RESULTS: Follow up ranged from four to 14 months. There was no recurrence of pterygium. Side effects of treatment included: delayed wound healing, avascularised sclera and pyogenic granuloma. CONCLUSION: This study suggests that intraoperative application of 0.5 mg/ml mitomyin C with 20 mg depot steroid following primary pterygium excision is effective and safe in preventing recurrence. PMID- 11486475 TI - Neuropathic pain--from mice to men. AB - The past decade has seen great progress in understanding the syndrome of neuropathic pain, its causes and in finding new drugs that promise great benefit. For example, an early outcome of the research has been the observation that the new drugs do not blunt normal pain sensation--a pattern beginning to find explanation through the realisation that neural pain circuits rewire themselves, both anatomically and biochemically, after nerve injury. In this article, we discuss a case of a known diabetic patient with intractable pain and the course of management provided by the use of novel tools and devices coming to the fore in this rapidly expanding specialty. PMID- 11486476 TI - [Leading symptoms of chest pain in the emergency room. Using cardiac markers for risk stratification]. AB - BACKGROUND AND OBJECTIVE: The acute coronary syndrome (ACS)--acute infarction or unstable angina pectoris--requires special monitoring and differentiated treatment. A prospective trial was undertaken to determine (1) clinical characteristics of patients with chest pain; (2) value of cardiac markers troponin T, myoglobin and CK-MB mass in differentiating cardiac and noncardiac chest pain; (3) the proportion of patients with ACS in whom these markers provided helpful additional information on admission and afterwards. PATIENTS AND METHODS: 233 consecutive patients (134 men, 99 women; aged 18-98 years), presenting in 1998 with nontraumatic chest pain at the emergency unit of the Lippe-Detmold Hospital were prospectively enrolled in the study. Levels of troponin T, myoglobin, CK-MB mass and activity and creatine kinase were measured on admission and after 2, 4 and 12-24 hours. RESULTS: ACS was ultimately diagnosed in 73 patients (31%). These were significantly older (p = 0.0015) and more often male (p = 0.046). 91% of patients with ACS but only 68% of those without described the chest pain as a sense of pressure or stabbing (p = 0.0002). Measurement of troponin T and myoglobin on admission was helpful in 39% of those with ACS, compared with 15% of those without. When the admission ECG was normal or nonspecific in patients with ACS, troponin T was elevated in 16%, CK-MB mass in 24%. In patients with unstable angina no single marker was associated with an increased risk (high rate of special supervision and intervention). CONCLUSION: In the assessment of patients with acute chest pain clinical judgement plays the predominant role. In the mostly elderly and male patients with ACS (31% of the cohort) feeling of pressure or stabbing chest pain were most prominent (91%). Cardiac markers troponin T, CK-MB mass and myoglobin were helpful in the differential diagnosis of chest pain, even when the ECG was unremarkable or nonspecific. At the time of admission myoglobin was the most significant marker for acute myocardial infarction or unstable angina. PMID- 11486477 TI - [A rare cause of malignant ascites--peritoneal mesothelioma]. AB - HISTORY AND CLINICAL FINDINGS: A 56-year-old patient (case 1) with recurrent haemorrhagic ascites for one year was admitted to our hospital for further investigation. Besides massive ascites he did not show abnormal physical signs. In addition, two 45-year-old patients were admitted (case 2 and 3) with clinical signs of acute abdomen--one having muscular guarding in the epigastric angle, the other in the right lower quadrant. All 3 patients did not have serious illnesses in the past; the first 2 patients had occupational asbestos exposure. INVESTIGATIONS: In patient 1 the ultrasound did not reveal abnormal findings besides ascites. Patients 2 and 3 underwent explorative laparotomy. DIAGNOSIS, TREATMENT AND COURSE: In the first case a diagnostic laparoscopy revealed diffuse tumor proliferations with nodular formations over the entire peritoneum- histologically a malignant peritoneal mesothelioma of the epithelial subtype. Patient 2 showed intraoperatively metastatic spread of tumour formations with infiltration of the peritoneum and transverse mesocolon. The histologic finding was similar to that in the first case. Patient 3 had a perforated sigma diverticulitis which was treated by resection of the sigmoid. Incidentally a well differentiated papillary peritoneal mesothelioma was found in the resected specimen. The first two patients were treated with alpha-interferon subcutaneously resulting in a decrease of ascites production. Because patient 3 showed neither ascites nor evidence for malignancy no interferon was administered. CONCLUSION: In case of haemorrhagic ascites of unknown cause a histological clarification by either laparoscopy or laparotomy is mandatory. Immunomodulation with interferon may be a promising approach. PMID- 11486478 TI - [Chronic hepatitis. 1: Diagnosis]. PMID- 11486479 TI - [Chronic hepatitis. 2: Treatment]. PMID- 11486480 TI - [The prevention of osteoporosis]. PMID- 11486481 TI - [Osteodensitometry in osteoporosis: the limits are wrong!]. PMID- 11486482 TI - [Osteodensitometry in osteoporosis: inadequate for diagnosis]. PMID- 11486483 TI - Neonatal 5,7-DHT lesions cause sex-specific changes in mouse cortical morphogenesis. AB - Both monoaminergic and cholinergic afferent projections to the neocortex putatively modulate cortical morphogenesis and plasticity. Previously we showed that neonatal electrolytic lesions of the cholinergic nucleus basalis magnocellularis (nBM) projections to the neocortex result in significant decreases of cortical layer width that correlate with cognitive alterations. Such electrolytic lesions, performed for lack of a selective neurotoxin in mice, may affect mono-aminergic fibers of passage. Here, we investigate the effects of neonatal 5,7 dihydroxytryptamine (5,7-DHT) focal injections into the nBM region on cortical laminar morphology in adult male and female mice. 5,7-DHT lesions on the first postnatal day resulted in significant cortical depletion of both serotonin and norepinephrine that attenuated with age. Generally, cortical layer widths increased in response to the lesion; the effects were layer, region, and sex specific. Previous reports from our laboratories described long-term behavioral alterations after comparable focal, neonatal 5,7-DHT lesions. The studies described here provide an anatomical basis for such behavioral alterations. Our data suggest that monoaminergic and cholinergic projections to the cortex may have opposite effects on the developing cortical neuropil. Jointly, our morphological and behavioral findings may have important implications for a variety of developmental disorders in humans and provide some insights into sex differences in the penetrance of these disorders. PMID- 11486484 TI - Parental deprivation induces N-methyl-D-aspartate-receptor upregulation in limbic brain areas of Octodon degus: protective role of the maternal call. AB - An optimal coordination between parents and their offspring involves a sequence of reciprocal behaviors to ensure the adequate care of the young, which is critical for a healthy physical, emotional, and social development. Parental deprivation, particularly an impaired child-mother attachment, induces lasting changes in emotional as well as in cognitive capacities in later life. We assessed in the South American precocial species, Octodon degus, whether alterations of glutamatergic function of the limbic system induced by parental deprivation may be a neural correlate for such behavioral changes. Further, we analyzed whether the mother's voice can protect from separation-induced changes of brain function. Changes of N-methyl-D-aspartate (NMDA) receptor expression were examined in the following three groups of 2-week-old Octodon degus pups: (I) control animals who remained undisturbed with the family; (II) animals who were repeatedly separated from the family and individually placed in an unfamiliar environment for behavioral analysis (open field); and (III) animals who were treated like the group described under (II) but were presented with maternal vocalizations during separation. Relative to those in the control group I, the animals in group II showed an upregulation of NMDA receptor density in the (a) anterior cingulate, prelimbic, infralimbic, and anterior insular cortices; (b) CA1/stratum radiatum; (c) CA1/stratum lacunosum moleculare and CA3/stratum radiatum; and (d) in the basomedial amygdaloid nucleus. Presentation of the maternal call during the separation period (group III) suppressed the separation induced NMDA receptor upregulation in all regions. The results demonstrate that early life events can influence the expression of transmitter receptors and that maternal behavior, acting to control the pup's socio-emotional environment, is a key factor for regulating such developmental events. PMID- 11486485 TI - Improvement of impaired memory in mice by taurine. AB - Taurine was extracted from Pegasus laternarius Cuvier to study its effects on learning and memory in mice. Mice were treated with different doses of taurine (10 mg/kg, 20 mg/kg, 40 mg/kg). The mice were treated with various chemical agents (pentobarbital, cycloheximide, sodium nitrite, alcohol) to disrupt the normal memory process. We measured the effect of taurine on step-down latency (SDL) and escape latency (EL) in a passive avoidance task after 10 or 30 days. Treatment with taurine alone did not change either SDL or EL. Taurine protected mice from the memory disruption induced by alcohol, pentobarbital, sodium nitrite, and cycloheximide but had no obvious effect on motor coordination, exploratory activity, or locomotor activity as measured using the rota-rod test and the hole board test. We conclude that taurine can be effective in attenuating the amnesia produced by alcohol, pentobarbital, cycloheximide, and sodium nitrite without compromising the behavioral aspects of the animals tested. PMID- 11486486 TI - Enhanced behavioral recovery from sensorimotor cortex lesions after pyramidotomy in adult rats. AB - Unilateral transection of the bulbar pyramid, performed before the ablation of the ipsilateral sensorimotor cortex, has been shown to facilitate the recovery of operantly conditioned reflexes and compensatory processes in rats. Such enhanced behavioral recovery was absent when only the sensorimotor cortex was ablated. This phenomenon is explained by the switching of motor activity under the control of the cortico-rubrospinal system. Switching of the descending influences is accomplished through the following loop: cortico-rubral projection-red nucleus inferior olive-cerebellum-thalamus-cerebral cortex. This suggests that a preliminary lesion of the peripheral part of the system, represented by a descending spinal projection, facilitates the recovery processes to develop during the subsequent destruction of its central part. PMID- 11486487 TI - CREB phosphorylation coincides with transient synapse formation in the rat hippocampal dentate gyrus following avoidance learning. AB - Spine density change in the hippocampal dentate gyrus accompanies memory consolidation and coincides with the increased expression of ribosome-rich, hyperchromatic granule cells. Although this suggests increased protein synthesis to be required for synaptic growth in the 5 to 7 h post-training period, little temporal mapping of the associated molecular mechanisms has been done. Here, we demonstrate a similar frequency of hyperchromatic cells in naive animals and in those sacrificed 6 h post-training, suggesting a transient repression of protein synthesis in the early post-training period. Immunoblot analysis of CREB phosphorylation in the dentate gyrus supported this view, with downregulation from basal levels observed at 2 to 3 h and at 12 h post-training. Protein synthesis reactivation appears to be specific for de novo spine production as no change in spine frequency accompanies the immediate post-training period of depressed protein synthesis. These findings support the view that CREB-mediated gene transcription is a requirement for long-term memory consolidation and may be directly implicated in the process of synaptic growth. PMID- 11486488 TI - ACTH4-10, substance P, and dizolcipine (MK-801) accelerate functional recovery after hemilabyrinthectomy in goldfish. AB - In this study, we evaluated the goldfish model of hemilabyrinthectomy for investigating potential recovery-promoting drugs. In this lesion model, the unilateral removal of the labyrinth induces a postural imbalance in response to light (Dorsal Light Reflex), from which the animals can recover over time. The behavioral effects of two neuropeptides were tested--namely, of substance P and ACTH4-10, both of which are known to promote functional recovery in several other lesion models. Furthermore, the effect of MK-801, an antagonist of the glutamatergic NMDA-receptor subtype, was tested because this substance has also been shown to exert a neuroprotective effect. After lesion of the right labyrinth, the animals (n = 12) were treated intraperitoneally daily either with vehicle (n = 12), substance P (n = 11), ACTH4-10 (n = 12), or MK-801 (n = 12). Another group (n = 11), which served as a non-lesion control, did not receive hemilabyrinthectomy or systemic injections. The lesion group, treated post operatively with vehicle, did not recover from the postural deviation over the 24 d testing period. In contrast, all three test substances accelerated the functional recovery after unilateral labyrinthectomy. The decrease of the dorsal light reflex persisted even after cessation of drug treatment after 20 d. The results indicate that using the dorsal light reflex in the model of hemilabyrinthectomy in goldfish provides a useful approach to studying the ability of potential new neurotrophic or neuroprotective drugs to promote functional recovery. PMID- 11486489 TI - Multiple-beam X-ray interferometry for phase-contrast microtomography. AB - The first successful operation of an X-ray interferometer under conditions of non planar three-beam diffraction for phase-contrast X-ray microtomography is reported. Intrinsic phase differences of the reflections used cancel from the three-dimensional phase image of the specimen. With simultaneous hkl and hkl reflections of a synchrotron radiation beam in a side-by-side geometry, the size of the usable field of view is doubled and the investigated specimen volume is increased by a factor of four. As an example, the reconstructed slice of a mouse kidney is shown in phase contrast at 71 keV. Optimized choices of three-beam reflections and matching interferometer geometries useful for applications are presented. PMID- 11486490 TI - High-pressure single-crystal X-ray diffraction facilities on station 9.8 at the SRS Daresbury Laboratory--hydrogen location in the high-pressure structure of ethanol. AB - A new high-pressure single-crystal diffraction facility has been constructed on station 9.8 at the Synchrotron Radiation Source, Daresbury Laboratory, for a range of studies on a variety of systems of relevance to physics, chemistry and materials science that would otherwise prove intractable with conventional laboratory-based methods. The station has been equipped with a modified Enraf Nonius CAD4 four-circle diffractometer for high-pressure studies which can be conveniently, and rapidly, interchanged with the Bruker SMART CCD area-detector system when more routine ambient-pressure diffraction work is to be undertaken. This rapid change-over has been achieved by permanently mounting the CAD4 on its own jacking table, formerly used for the station's white-beam diffraction mode, which allows the alignment of the SMART diffractometer to remain undisturbed when the CAD4 is in use. Early results on the test low-melting-point compound ethanol (CH3CH2OH) reveal that excellent refined structures can be obtained, including the location and refinement of the H atoms, demonstrating that one of the original, and major, objectives of the station has been accomplished. PMID- 11486491 TI - Fixed-height exit bender of synchrotron X-rays above 40 keV. AB - A crystal bender for sagittal focusing has been designed for standard monochromators at SPring-8. The bender does not move the position of the crystal center when the bending radius is changed. Sagittal focusing from 40 keV to 60 keV was achieved by using Si(311) double crystals. The flux density of the focused beam measured at 40 keV was 15 times higher than that of the unfocused beam. The height deviation of the focused beam throughout the measured energy range was within +/- 0.15 mm. PMID- 11486492 TI - Performance limits of indirectly cryogenically cooled silicon monochromators- experimental results at the APS. AB - The results of high-heat-load tests of indirectly cryogenically cooled silicon monochromators are presented. The measurements show that, provided that the total power absorbed by the crystal is less than approximately 150 W, indirect cryogenically cooled silicon monochromators will perform well, with thermal induced slope errors of less than 2 arcsec. At the Advanced Photon Source, this corresponds to the undulator closed-gap (11 mm) condition at 100 mA with white beam slit sizes slightly larger than the full width at half-maximum of the radiation central cones. The dependence of the slope errors on the thermomechanical properties of silicon are discussed and clearly demonstrated. PMID- 11486493 TI - A new X-ray spectrometer for high-resolution Compton profile measurements at SPring-8. AB - An X-ray spectrometer for high-resolution Compton profile measurements using 90 120 keV X-rays has been designed and constructed at SPring-8. A Cauchois-type triply layered bent-crystal analyzer was employed for the energy analysis. A novel use of a solid-state detector with a large active area was devised as a position-sensitive detector. A resolution of 0.10 atomic units in electron momentum has been achieved at an incident X-ray energy of 115 keV. A Compton profile of a single crystal of Nb was measured with a counting rate of 30 counts s-1 at the Compton peak, which demonstrates that the spectrometer is capable of measuring Compton profiles of heavy-element materials. PMID- 11486494 TI - X-ray double phase retarders to compensate for off-axis aberration. AB - An X-ray double phase retarder system composed of two transmission-type phase retarders is proposed and developed in order to compensate for off-axis aberration (phase-shift inhomogeneity due to angular divergence of incident X rays). The scattering planes of the two phase retarders are set to be inclined by 45 degrees with respect to the plane of incident polarization, but the two phase retarders give Bragg reflections in opposite directions. By using this X-ray optical system, vertically polarized X-rays with a 0.99 degree of linear polarization were obtained from horizontally polarized synchrotron radiation with a horizontal beam divergence of 20 arcsec (0.1 mrad). This value is favorably compared with the value of 0.87 which was obtained using a conventional single phase retarder of identical total thickness, 627 microns. The comparison was made at the nickel K-absorption edge (8333 eV) with the condition that 47% of incident X-rays were transmitted through the two phase retarder crystals. The crystals were (100)-oriented diamond plates giving asymmetric 111 Laue reflections. PMID- 11486495 TI - Piezo-QEXAFS with fluorescence detection: fast time-resolved investigations of dilute specimens. AB - First X-ray absorption spectroscopy experiments with a vibrating piezo-driven double-crystal monochromator (piezo-QEXAFS) and fluorescence detection are reported. It is shown that high-quality XANES spectra can be recorded on a time scale of about 50 ms per spectrum, even for very low concentrations of < 10 mmol l-1 using fluorescence detection. The quality of the spectra, possible applications, as well as present limits of the technique will be discussed. PMID- 11486496 TI - [Perspectives in the treatment of hypertension in primary care]. AB - Hypertension control by primary care practitioners has improved but is still not satisfactory. Four dilemmata seem to prevent further progress: pharmacological pseudo-innovations, discordance with regard to treatment objectives between patients and doctors, professional heteronomy and limited scope for behavioural change. With ample evidence from epidemiological and intervention studies being available, primary care practitioners are now in a position to counsel their patients more effectively. Models of shared decision-making and motivational interviewing will help to establish a new paradigm of care. However, high-risk approaches aiming at individual risk factor modification are not sufficient. Causes of high blood pressure and cardiovascular morbidity that operate at the population level and limit individual prevention have to be explored and modified. PMID- 11486497 TI - [Guideline-adequate knowledge in internists and general practitioners about the diagnosis and treatment of arterial hypertension]. AB - Only a small proportion of patients with arterial hypertension are adequately treated. Although a possible cause for this fact may be the deficient knowledge of physicians about diagnosis and treatment of arterial hypertension, to date no studies have addressed this important problem in Germany. Therefore, we have reviewed the knowledge of internists and general practitioners about diagnosis and treatment of arterial hypertension using a questionnaire. The guidelines of the German Hypertension Society were used as the reference standard. The questionnaire was sent out in December 1999 to all internists (n = 15,952) and to a random sample of general practitioners (n = 8947) who work as statutory health insurance physicians. A total of 11,547 questionnaires were sent back and could be analysed. Adequate guideline knowledge was assumed if five out of eight questions were correctly answered; the correct answers had to include the correct definition of arterial hypertension (> 140/90 mmHg). The correct definition of arterial hypertension was known by 4103/11,547 participants (36%). An adequate guideline knowledge was found in 18.8% of the general practitioners and in 26.6% of the internists. There were no relevant regional differences. The level of awareness about diagnosis and treatment of arterial hypertension is insufficient among internists and general practitioners. This insufficient knowledge may in part explain the inadequate care for patients with arterial hypertension. Thus, the implementation and evaluation of new information and training strategies are mandatory to improve the care for patients with arterial hypertension. PMID- 11486498 TI - [Pharmacotherapy of hypertension]. AB - A large variety of drugs is available for treatment of hypertension. Moreover, many randomised controlled trials with clinically relevant endpoints (morbidity, mortality, quality of life) do exist in the cardiovascular field, providing for sufficient evidence to choose the appropriate agent in most circumstances. For diuretics and betablockers a large body of evidence in terms of beneficial effects on outcome does exist, for ACE-inhibitors in some special indications only. These drugs are therefore recommended as first-line treatments. For calcium channel blockers (with the exception of isolated systolic hypertension in the elderly) and AT1-receptor-antagonists the results of endpoint-studies are still awaited. These results will have to be considered for revised versions of currently available guidelines. PMID- 11486499 TI - [Optimized therapy for hypertension]. AB - The benefit of good blood pressure control in patients with arterial hypertension has been repeatedly demonstrated. In high-risk patients, a good blood pressure control is one of the main prerequisites for the reduction of morbidity and mortality. However, as has been shown in population-based studies, the quality of blood pressure control both in patients with essential hypertension and in hypertensive diabetic patients is still unsatisfactory. Only a minority of patients achieve target blood pressure values below 140/90 mm Hg. This situation has not changed within the recent years, although the prescriptions of antihypertensive drugs have continuously increased. This paper describes a structured hypertension treatment and teaching program (HTTP) which was developed and evaluated during the last 16 years and which aims at intensification of antihypertensive therapy by active involvement of the patients in their own treatment. This programme primarily focuses on the improvement of the patients' long-term treatment compliance by means of information about non-pharmacological and pharmacological therapies and instructions to regular blood pressure self measurements. The efficacy and feasibility of the HTTP has been evaluated in several studies which unanimously show considerable benefits not only concerning blood pressure control but, most importantly, regarding reduction in the incidence and progression of hypertension-associated diseases as cerebro- and cardiovascular events and diabetic nephropathy. Therefore, it seems essential to implement the HTTP in the German health care system in order to finally achieve the desired quality of care for patients with arterial hypertension. PMID- 11486500 TI - [Age-related macular degeneration--a public health problem]. AB - Verteporfin for the treatment of age related macular degeneration was identified as a potential breakthrough technology by the Working Group on Medical Procedures of the German Standing Committee of Physicians and Sickness Funds in spring 2000. Consequently, a formal review was undertaken to assess the benefits and risks of this treatment in order to evaluate its future status in German statutory ambulatory health care. The treatment effect of verteporfin was established in a high quality randomized controlled trial, for the predominantly wet, classic type of choroidal neovascularization secondary to age related macular degeneration. For this patient group, verteporfin is regarded as a reasonable and necessary procedure as there is no effective alternative therapy for this type of macular degeneration. However, the treatment stabilizes the patient's vision rather than improves it and the long term effects must still be evaluated in further clinical trials. After a thorough review of the scientific literature and the statements of various stakeholders the Standing Committee decided to introduce this technology into the statutory healthcare benefits package. For the most effective use of this treatment it must be assured that, prior to treatment, the AMD type (wet vs. dry, classic vs. occult) is correctly diagnosed (using fluorescein angiography) and documented by specially trained physicians. For this reason a stringent quality assurance guideline was developed to prevent the use of this costly technology for non-approved indications. Further indications for this therapy are currently under evaluation in randomized controlled trials. Hence, this decision will need to be updated in future. PMID- 11486501 TI - [Selection, problems and perspective of quality indicators]. AB - The advances in information technology offer new possibilities to gather and analyse health-related information in order to get an inside view into the quality of medical care. Applied medical interventions and the related outcome for the patient can be evaluated if information of a certain degree of complexity is available. Performance measures are based on collection and aggregation of data. They try to abstract medical performances in order to get objective and comparable variables. These variables are useful in evaluating and monitoring the different dimensions of patient care. Performance measurement can inform people about the outcomes they can expect from certain treatments. Beside formal criteria such as validity and reliability, the specific objectives determine the selection of a relevant performance measure. This article describes actual problems and solutions in the data collection and the analysis of performance measures for the domain of oncology. This way, we want to encourage the meaningful, but critical use of performance measures in medicine. The provision of such information can build the basis for the process of medical decision making by physicians, the selection of a suitable medical institution by patients, and the allocation of resources by providers. PMID- 11486502 TI - [Development of quality assurance criteria for continuing education in evidence based medicine]. AB - OBJECTIVE: To develop a set of core contents for teaching evidence-based medicine (EBM). METHOD: Description of basic principles from clinical epidemiology, medical statistics and literature search essential for health care practitioners to perform critical appraisal of the medical literature. Based on textbooks for EBM and clinical epidemiology and guided by the EBM courses at McMaster university and the Oxford Centre for EBM, core contents were defined. Decision on inclusion was made by consensus. RESULTS: The core contents for the Berlin curriculum in EBM covers 3 areas: 1. General information on the methods of experimental and observational clinical studies, principles of EMB, basic statistical concepts, and literature search. 2. Tools for critical appraisal of primary studies required for medical decision making: therapy, diagnosis, prognosis, and harm. 3. Evidence-based summary reports: meta-analysis and guidelines. CONCLUSIONS: The contents of EBM can be precisely defined. They are transparent, explicit and communicable and present a reproducible basis for the different forms of teaching EBM. PMID- 11486503 TI - [Health consequences of passive smoking]. AB - Environmental tobacco smoking (ETS) represents a main risk factor for the generation of diseases of the respiratory tract and of the cardiovascular system in spite of statements to the contrary. ETS enhances the risk of lung cancer by a factor of 2-3. Newborn and small children (< 2 years of life) are at high risk if they live within this period of time in a household exposed to maternal more than fraternal smoking. Endothelial cells of the blood vessels are damaged as early as during the first month of life of passive smoking children, and these defects can be detected during the first decade of life. ETS over a period of more than ten years changes the intima/media ratio by enhancing the thickness of the vessel wall. Additionally, poor health behaviour is seen in households of smokers because the behaviour of the parents is transferred to that of their children, and this behaviour is the starting point of further health risks and damages. The presented data should cause a call for primary smoking prevention preferably among children and young persons on the one hand, and the organisation of programs against ETS at the workplace and in public buildings, as well as in the private house on the other hand. Non-smokers must be informed about the risks and dangers of ETS more than it is the case up to now. PMID- 11486504 TI - Molecular diversity, structure and domestication of grasses. PMID- 11486505 TI - Imprinting of R-r, paramutation of B-I and Pl, and epigenetic silencing of MuDR/Mu transposons in Zea mays L. are coordinately affected by inbred background. AB - The extent of imprinting at R-r, frequency of paramutation at B-Intense and Pl, and epigenetic silencing of Mu transposons were evaluated in the W23 and A188 inbred lines of maize. All types of epigenetic phenomena affecting these loci of the anthocyanin pathway occurred more frequently in the W23 inbred line. Absence of down-regulation was dominant in F1 hybrid progeny. Identical alleles programme lower anthocyanin accumulation in A188 than in W23, and A188 plants develop more rapidly than W23. The possibilities that specific genetic factors, intrinsic gene expression levels and/or the rapidity of the life cycle modulate epigenetic gene controls are discussed. PMID- 11486506 TI - Chromosomal distribution of the transposable elements Osvaldo and blanco in original and colonizer populations of Drosophila buzzatii. AB - Chromosomal distribution of transposable elements (TEs) Osvaldo and blanco in D. buzzatii was studied in three original natural populations from Argentina (Berna, Puerto Tirol and La Nostalgia) and a colonizer population from the Iberian Peninsula (Carboneras). The Spanish population showed significant differences for Osvaldo and blanco copy numbers when we compared the X chromosome and the autosomes; but it is mainly the accumulation of copies in chromosome 2, where most sites with high insertion frequency were located, that causes the discrepancy with the negative selection model. We found no significant differences in TE frequency between chromosomal regions with different exchange rates, and no evident accumulation of TE was detected within chromosomal inversions where recombination rate is reduced. The Carboneras population shows euchromatic sites of Osvaldo and blanco with high occupancy and others with low copy number. On the contrary, Argentinian populations show only a generalized low occupancy per insertion site. Moreover, the mean copy number of both elements is higher in Spain than in Argentina. All these results suggest an important role of the colonization process in the distribution of TEs. The increase in the copy number of the TEs analysed and their elevated frequency in some chromosomal sites in Carboneras is, most probably, a sequel of the founder event and drift that took place at the time of the colonization of the Old World by D. buzzatii from the New World some 300 years ago. PMID- 11486507 TI - PGal4 excision reveals the pleiotropic effects of Voila, a Drosophila locus that affects development and courtship behaviour. AB - In Drosophila melanogaster, the PGal4 transposon inserted at the chromosomal site 86E1-2 is associated with the Voila1 allele that causes multiple phenotypes. Homozygous Voila1/1 flies rarely reach adulthood and heterozygous Voila1/+ adult males display strong homosexual courtship behaviour. Both normal behavioural and developmental phenotypes were rescued by remobilizing the PGal4 element. Yet, the rescue of heterosexual courtship and of adult viability did not occur in the same strains, indicating that these defects have different genetic origins. Furthermore, many strains showed a partial rescue of both characters. Molecular analysis revealed that the PGal4 transposon is inserted upstream of the 5'UTR of the prospero gene. The excision strains with no detectable fragment of the PGal4 transposon remaining showed a rescued viability for homozygote adults. Moreover, the developmental period with the highest homozygote lethality was correlated with the size of PGal4 element that remained inserted at the Voila locus. This suggests a relationship between developmental viability and the amount of DNA inserted within the promoter of prospero. PMID- 11486508 TI - Microsatellite isolation, linkage group identification and determination of recombination frequency in the peach-potato aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae). AB - Fifteen polymorphic microsatellite markers were used to establish linkage groups and relative rates of recombination in male and female Myzus persicae (Sulzer) (Hemiptera: Aphididae) (peach-potato aphid). We cloned nine markers from M. persicae and for these we report primer sequences and levels of allelic diversity and heterozygosity in four Australian M. persicae populations. Of the remaining six loci, four loci, also cloned from M. persicae, were obtained from G. Malarky (Natural History Museum, London) and two loci from Sitobion miscanthi were used. Additionally, the primer sequences of locus M77, a locus monomorphic in M. persicae but polymorphic in the closely related Myzus antirrhinii, are presented. Eleven of the 15 polymorphic markers were autosomal and four were X-linked. A linkage analysis was performed on a European pedigree of aphids containing five families with between seven and 11 offspring each. There was no linkage between any loci in females. In males, several pairwise comparisons yielded no recombinant offspring. With the exception of locus M40, these observations were supported in a linkage analysis performed on larger families produced from Australian M. persicae crosses. Locus M40 showed segregation consistent with involvement in a translocation between autosomes 1 and 3 in European samples but not in the Australian samples. From the Australian crosses we report an absence of recombination in males but high recombination rates in females. One X chromosome and four autosomal linkage groups were identified and tentatively assigned to chromosomes. The relevance of achiasmate meiosis to the evolution of sex is discussed. PMID- 11486509 TI - Transposable element number in mixed mating populations. AB - Theoretical population genetic studies of transposable elements focus almost exclusively on random mating species, whereas many plants reproduce through partial or substantial self-fertilization. Here I develop computer simulation and analytic approximations of simplified element dynamics (transposition balanced by selective elimination) in partially self-fertilizing populations, using Ty1-copia elements for biological inspiration. Under the most plausible models and parameter values, element numbers decrease with self-fertilization when element insertions are deleterious, but may increase when ectopic exchange regulates element number. Conclusions for models of ectopic exchange depend in part on parameters for which little firm empirical evidence is available. Small changes in selfing rate can lead to abrupt changes in element number when homozygous and heterozygous elements have markedly different fitness effects. Equilibrium element numbers can be sensitive to population size, especially at high selfing rates. Elements are frequently lost in small highly selfing populations under the deleterious insertion model. In contrast, small highly selfing populations can accumulate very large numbers of elements under ectopic exchange. Empirical data on element number and localization in plants with different mating systems suggests that deleterious insertion, rather than ectopic exchange, may regulate element number. Limitations to available empirical data, especially the lack of comparison between closely related species differing in mating system, mean that this conclusion is tentative. PMID- 11486510 TI - Reproductive compensation and human genetic disease. AB - The effects of reproductive compensation on the population genetics of sex-linked recessive lethal mutations are investigated. Simple equations are presented which describe these effects, and so complement existing population genetic theory. More importantly, this type of mutation is responsible for several severe human genetic diseases such as Duchenne muscular dystrophy. It is argued that the applications of three modern reproductive technologies--effective family planning, in utero diagnosis with termination, and embryo sexing--will lead to reproductive compensation. The adoption of any of these technologies may rapidly elevate the frequencies of those mutations which are lethal in childhood. This increase is large, in the order of 33% upwards, and occurs rapidly over two to five generations. It also depends on the source of mutations, the effect being larger if most mutations are paternal. In utero diagnosis and/or embryo sexing increase the frequency of the mutation, but simultaneously decrease disease incidence by preventing the birth of affected offspring. In contrast, effective family planning may rapidly increase both mutation frequency and disease incidence. PMID- 11486511 TI - Minimization of rate of inbreeding for small populations with overlapping generations. AB - We propose a method that minimizes the rate of inbreeding (delta F) for small unselected populations with overlapping generations and several reproductive age classes. It minimizes the increase in coancestry of parents and optimizes the contribution of each selection candidate. The carrying capacity of the population is limited to a fixed number of animals per year. When survival rate equalled 100%, only animals from the oldest age class were selected, which maximized the number of parents per generation, slowed down the turnover of generations and minimized the increase of coancestry across sublines. However, the population became split into sublines separated by age classes, which substantially increased inbreeding within sublines. Sublines were prevented by a restriction of selecting at least one sire and one dam from the second-oldest age class, which resulted in an L times lower delta F, where L equals the average generation interval of sires and dams. Minimum coancestry mating resulted in lower levels of inbreeding than random mating, but delta F was approximately the same. For schemes where the oldest animals were selected, delta F increased by 18-52% compared with the proposed method. PMID- 11486512 TI - [Repair and revision 5. Failures and repair of resin-bonded bridges]. AB - This article describes a number of aspects dealing with debonding, recementation and patients' satisfaction in the treatment with resin-bonded bridges. Experiences from two large clinical trials are reported. It was found that rebonded bridges were less successful than the originally bonded bridges. However, since the functional survival rate was significantly higher than the original survival rate, together with the relative low cost of recementation, it is advised to recement debonded bridges. After multiple debonding recementation can be questioned. Multiple debonding is considered to point to an inadequate indication of this type of fixed prostheses. Debonding of bridges did not alter the patients' satisfaction. This might be caused by the fact that the possibility of failure was explained to the patient when informed consent was obtained. PMID- 11486513 TI - [Meta-analysis of anterior veneer restorations in clinical studies]. AB - A systematic review was used to make an inventory of clinical studies on veneer restorations. Based on a protocol articles were selected that were subsequently assessed for their quality. With 'failure of the restoration' as a study outcome, survival data of the restorations were generated from the selected studies and a mean survival result was composed. Porcelain veneer restorations showed an acceptable 3-year survival, which was favourable to the 3-year survival of preformed acrylic veneers. The quality assessment expressed that reports on clinical studies should be more standardised in order to make it easier to compare the results of different studies. PMID- 11486514 TI - [Meta-analysis and 'evidence-based practice' in dentistry]. AB - The dental practitioner may sometimes search for certainty in dental decisions from the dental literature. In this 'Evidence Based Dentistry' situation a meta analysis, as a structured and statistically supported literature review, may be helpful. The papers that are included in the meta-analysis have to be carefully screened and selected for a correct clinical decision. PMID- 11486515 TI - [Sterilization of dental instruments]. AB - Instruments used by dentists are often sterilized with steam sterilization. At this moment three types of processes are described in the European norms: Type N, S and B. According to the norms type N can be used for non-wrapped non-hollow instruments and non-porous instruments, type S can be used for instruments specified by the manufacturer and type B can be used for wrapped, hollow and porous instruments. The principles on which the sterilizors are based are described. It is concluded that before purchasing the sterilizor the method of working and instruments to be sterilized must specified. In most cases a type B process is preferable in order to gaurantee to be certain to have an effective and reproducible process. PMID- 11486516 TI - [Validity of a new caries indicator dye in detection of carious, infected dentine]. AB - During cavity excavation diagnostic test are used to prevent any unnecessary removal of sound dentine. In this study the validity of a newly introduced blue dye (Quadrant CariTest) was investigated. Twenty-five carious teeth were collected immediately after extraction. The caries lesions were stepwise excavated. From four excavation steps per lesion a digital and an ordinal blue measurement was obtained. From each excavation phase a little amount of excavated dentine was cultured on blood plates. The correlation coefficients between the ordinal and the digital blue measurements and the counts of micro-organisms, expressed in log(CFU)/ml, were respectively 0.56 and 0.45. The sensitivity and the specificity of the ordinal blue measurements were 0.69 and 0.83, those of the digital blue measurements 0.76 and 0.65. The area under ROC of the ordinal blue measurements was 0.77, and that of digital blue measurements 0.70. The areas under ROC were not significantly different (p = 0.14). It was concluded that the new caries indicator is well applicable during the cavity preparation to demonstrate infected dentine. PMID- 11486517 TI - [Chemo-mechanical caries removal: a review of the techniques and latest developments]. AB - Chemo-mechanical caries removal involves the chemical softening of carious dentine followed by its removal by gentle excavation. The reagent involved is generated by mixing amino acids with sodium hypochlorite; N-monochloroamino acids are formed which selectively degrade demineralised collagen in carious dentine. The procedure requires 5-15 minutes but avoids the painful removal of sound dentine thereby reducing the need for local anaesthesia. It is well suited to the treatment of deciduous teeth, dental phobics and medically compromised patients. The dentine surface formed is highly irregular and well suited to bonding with composite resin or glass ionomer. When complete caries removal is achieved, the dentine remaining is sound and properly mineralised. The system was originally marketed in the USA in the 1980's as Caridex. Large volumes of solution and a special applicator system were required. A new system, Carisolv, has recently been launched on to the market. This comes as a gel, requires volumes of 0.2-1.0 ml and is accompanied by specially designed instruments. PMID- 11486519 TI - [Reactions to the seventh national practice guideline for pediatric dentistry]. PMID- 11486520 TI - [Extraction in the anterior mandible: should the patient be referred to oral surgeon?]. PMID- 11486521 TI - [Migraine and cluster headache 1]. PMID- 11486522 TI - [Problem-based dentistry on internet]. PMID- 11486523 TI - [Optimal sealing of fissures]. PMID- 11486524 TI - [What is your diagnosis. Diffuse idiopathic skeletal hyperostosis (DISH) with chronic, post-traumatic lesion of the 6th left rib with hyperostotic reaction within the scope of DISH]. PMID- 11486525 TI - [Illness, sick role and literature]. PMID- 11486526 TI - [Do physicians need fictional literature? Dialogue between an enthusiast and a skeptic]. PMID- 11486527 TI - ["Why does it hit me?" Illness as an existential question]. PMID- 11486528 TI - [Thoughts on the meaning of suffering]. PMID- 11486529 TI - [Goethe--medical awareness]. PMID- 11486530 TI - [Dostojewskij and epilepsy--a literary and epileptological observation]. PMID- 11486531 TI - [Somatoform pain disorder and work disability]. PMID- 11486532 TI - [Laparoscopic radical prostatectomy]. PMID- 11486533 TI - [Physiopathologic consequences of laparoscopic surgery]. AB - The aim of this present review was to study the effects of the intraperitoneal carbon dioxide insufflation, on the different physiological functions in order to prevent or to treat the side effects or complications which may occur in laparoscopic surgery. The major data from literature are confronted with the practical knowledge from a group of anesthetists with large experience in gynecological laparoscopic surgery. PMID- 11486534 TI - [Pancreas and kidney transplantation: long-term metabolic results]. AB - STUDY AIM: Pancreas and kidney transplantation (PKTx) is indicated in uremic patients with insulin-dependent diabetes mellitus (IDDM). The aim of this study was to determine its long-term effect on metabolic control in order to establish the real efficacy of this treatment in diabetic patients. PATIENTS AND METHOD: Among a total experience of 191 pancreas and kidney transplantations, a metabolic control was performed in 80 patients who underwent PKTx in our center, with both grafts functioning for more than one year. Immunological markers of diabetes mellitus were also evaluated (ICA and GADab) in 50 patients. RESULTS: Basal glycemia and glycosylated hemoglobin (HbA1c) levels throughout follow-up were within the normal range. Hyperinsulinemia was present throughout follow-up till the fourth year. The oral glucose tolerance test (OGTT) was normal in 82.5% of the patients beyond one year after the graft. Over time, no differences were detected on basal glucose and insulin levels and areas under the curve (AUC) of glycemia and insulinemia. During the evolution, no differences were found in the fasting insulin resistance index (FIRI), in spite of increasing body weight. ICA were + in 2 patients before graft and + in 7 after graft (14%). GADab were + in 10 patients before graft and + in 11 after graft (22%). CONCLUSION: Pancreas and kidney transplantation provides without any insulin treatment and diet long-term normalization of glycemic control, assessed by HbA1c and OGTT, despite the existence of sustained hyperinsulinemia. Our results strongly suggest that pancreas and kidney transplantation is the most efficient treatment for uremic patients with insulin-dependent diabetes mellitus from a metabolic point of view. PMID- 11486535 TI - [Esophagectomy for squamous cell carcinoma of the esophagus isolated or associated with head and neck cancer: long-term survival]. AB - STUDY AIM: Esophageal squamous cell carcinomas are frequently associated with head and neck cancers. This retrospective study was conducted to compare the long term outcome of the patients with a double cancer and of the patients with a solitary esophageal cancer after curative management. PATIENTS AND METHODS: From 1989 to 1999, 114 patients with an esophageal carcinoma were included in the study. Among them, 52 had an associated head and neck cancer (metachronous: n = 17 and synchronous: n = 35). Curative treatment was achieved in all patients. The patients were divided in "solitary" and "associated" group. RESULTS: Age, sex distribution, tumor location and histological findings were similar in the two groups. The esophageal resection was an esopharyngolaryngectomy (n = 13), a subtotal esophagectomy with cervical anastomosis (n = 92) and a Lewis-Santy esophagectomy with thoracic anastomosis (n = 9). Operative mortality (8 versus 7.7%), anastomotic leaks rate (14.5 versus 21%) and pneumonia rate (21 versus 9.6%) were not significantly different in the two groups. The mean hospital stay was 27 days. The mean follow-up was 85 +/- 50 months. Five-year survival rates were not significantly different in the two groups (p = 0.6411). In univariate survival analysis the only significant predictive factors were the depth of esophageal tumor invasion (p = 0.0002) and node involvement (p = 0.0373). The presence of head and neck cancer did not affect survival after esophagectomy. CONCLUSION: With an aggressive therapeutic plan, the survival of patients with an esophageal cancer associated to head and neck cancer was similar to the survival of patients with an isolated esophageal carcinoma. Long term esophageal survey seems to be useful to detect more superficial esophageal carcinomas in patients with head and neck cancer. PMID- 11486536 TI - [Radioguided surgery for primary hyperparathyroidism. Experience in 75 cases]. AB - STUDY AIM: To report feasibility and efficacy of radioguided mini invasive hyperparathyroidism surgery. PATIENTS AND METHOD: From November 1998 to August 2000, 75 patients with primary hyperparathyroidism have been operated on by radioguided surgery within 90 to 180 minutes following i.v. injection of a diagnostic dose of MIBI and after parathyroid scanning with planar and oblique views. Exclusion criteria were thyroid pathology requiring surgery and suspicion of multiple endocrine neoplasia. A 20% step-up between the background noise and tissue uptake was the diagnostic threshold for parathyroid hyperfunctioning tissue. RESULTS: A 20% step-up was observed in only 17% of cases overall; all cured, but two. Various angles of application of the probe can result in significant discrepancies of recorded uptake for the same spot. These results demonstrate a physiological step-up between the ipsilateral unaffected upper and lower quadrants of the neck (range: -17 to -8%), because of the proximity of supraortic vessels (upper neck-upper mediastinal gradient: -44 to -30%). Therefore, significant ratios are meaningful only between either the symmetrical left and right controlateral quadrants respectively, and not between the upper and the lower ipsilateral quadrants. Eleven per cent (8/75) of preoperative scannings were non-contributory, and probe detection was contributory in 3/8 cases only. CONCLUSION: Benefit of the technique is limited in routine, but it can be helpful in redo cases if the offending gland is not located in close surroundings of tissues physiologically taking up the radiopharmaceutical (salivary glands, great vessels and heart). PMID- 11486537 TI - [Abdominoperineal resection for locally recurrent rectal cancers following anterior resection]. AB - STUDY AIM: The aim of this retrospective study was to evaluate the short and long term results of abdominoperineal resection for local recurrence following low anterior resection of a rectal adenocarcinoma and to determine the prognostic factors. PATIENTS AND METHODS: From January 1978 to December 1996, 35 patients (17 women, 18 men) with a mean age of 59.4 years, underwent an abdominoperineal resection for local recurrence after low anterior resection of a rectal adenocarcinoma. The primary tumor was below the peritoneum in 29 cases, and the mean security margin was 3 cm under the tumor. Tumor staging at the time of primary surgery included 23 Dukes B, 11 Dukes C, and 1 Dukes D. The mean time elapsed between low anterior resection and local recurrence was 16.4 months. The histological diagnosis of recurrence was obtained preoperatively in 29 cases (82.8%). RESULTS: Resection was curative in 12 patients and palliative only in 23 patients. The recurrence was intramural in 3 cases, extramural in 10 cases, and mixed in 22 cases. Ten patients had an extended "en bloc" resection including one or several adjacent organs, and a synchronous metastasis was resected in 2 cases. The mortality rate was 2.8% (n = 1) and the morbidity rate was 23% (n = 8). The 1 year and 5-year survival rates were respectively 77 and 30.2% with the univariate analysis of prognosis factors of survival, there were four pretherapeutic factors (age, staging of the primary tumor, delay of the recurrence, CEA rate) and four therapeutic factors (curative resection, extramural recurrence, staging of the recurrence, postoperative radiotherapy). The curative or not curative type of resection was the only independent predictor of survival with multivariate analysis. CONCLUSION: The results of this study seem to justify an abdominoperineal resection for local recurrence after low anterior resection whenever possible. Long-term results may possibly be improved by using adjuvant treatment. PMID- 11486538 TI - [Composite, non-resorbable parietal prosthesis with polyethylene terephtalate polyurethane (HI-TEX PARP NT): prevention of intraperitoneal adhesions. Experimental study in rabbits]. AB - The authors report an experimental study in the rabbit with a new composite non absorbable mesh in Polyethylene Terephtalate-Polyurethane used for incisional hernia repair in intraperitoneal positioning. This new mesh has one permeable side in polyethylene terephtalate for rapid tissue fixation and another side in polyruethane, hydrophob in order to avoid cell penetration. Eighteen rabbits were operated. A wound was created in aponeurose, muscle and peritoneal abdominal wall. The mesh was placed in intraperitoneal positioning and was taken off at 4, 9 and 13 months for histologic examination and electronic microscopical examination. Tolerance, adhesion, tissular reaction and neoperitoneum formation have been studied. All the meshes were well integrated and without sepsis. In 18% of cases small and monocclusive intraperitoneal adhesions were found. This new composite mesh in intraperitoneal positioning gave good results at medium-term in the rabbit. It's an attractive alternative for incision hernias repair with intraperitoneal mesh. PMID- 11486539 TI - [Experimental use of laparoscopic material: report of a case of spondylodiscitis after laparoscopic sacropexy with Taker]. PMID- 11486541 TI - [Primary fibromatosis of the breast]. AB - Primary fibromatosis of the breast is a very rare pathology which looks like a malignant tumor, clinically and radiologically. Fibromatosis of the breast is characterised by a local invasion and a risk of recurrence. Positive diagnosis is provided by histology and its treatment is surgical. The aim of this study was to report one case and to emphasize the difficulties encountered in the diagnosis and the treatment. PMID- 11486540 TI - [Primary malignant melanoma of the esophagus]. AB - The primary malignant melanoma of the esophagus is a rare tumor. The study aim was to report two cases, one treated by esophagectomy without thoracotomy and the other one by Lewis-Santy type esophagectomy. Both patients had recurrence. One died at the 24th month with liver metastases. The other one who had a cervical invaded lymph node, treated by radio-chemotherapy, is actually in complete remission 9 years after the diagnosis. PMID- 11486542 TI - [A rare complication of ileal pouch anal anastomosis: superior mesenteric artery syndrome]. AB - The aim of this study was to report a new case of superior mesenteric artery syndrome following total proctocolectomy and ileal pouch-anal anastomosis for chronic ulcerative colitis. Diagnosis was made following a prolonged gastric stasis and was based on upper gastrointestinal X-ray series. Medical treatment was unefficient and gastro-jejunostomy was performed. The early outcome was good. PMID- 11486543 TI - [Revision of the Huriet-Serusclat law: position of medical and patient associations]. PMID- 11486544 TI - [Eugene Koeberle (1828-1915), founder of modern surgery]. AB - Eugene Koeberle, surgeon and anatomist gained international renown due to his outstanding ability in resection of ovary cysts and in hysterectomy. He was one of the first to perform these operations successfully. He was a pioneer in asepsis and pre- and postoperative care. He also invented many surgical instruments, among them an efficient hemostatic forceps. PMID- 11486545 TI - [Eulogy for George Cerbonnet (1922-2000)]. PMID- 11486546 TI - [Intestinal lipoma resulting in intestinal intussusception and anemia: ultrasonographic diagnosis and video-assisted treatment]. PMID- 11486547 TI - [Jejunal tuberculosis revealed by a perforation]. PMID- 11486548 TI - [Giant colonic diverticulum]. PMID- 11486549 TI - [Intestinal obstruction by migration of wire mesh into the intestinal lumen following hernia repair]. PMID- 11486550 TI - [Extremely large bursa of the knee]. PMID- 11486551 TI - [Communication of an hydatid cyst of the liver with the skin and the biliary tract and bronchi]. PMID- 11486552 TI - [Ethical bases for clinical studies]. PMID- 11486553 TI - [Fluoroquinolones--a current review]. PMID- 11486554 TI - [Headache: is there a connection with diet?]. PMID- 11486555 TI - [Anaphylactic shock: diagnosis and therapy]. PMID- 11486556 TI - Bilateral sixth-nerve palsy associated with dural arteriovenous malformation. AB - A fifty-two year old postmenopausal woman was admitted to the hospital with complaints of diplopia, headache, humming over the head, and pain over the left eye. Neurological examination showed right abducens nerve palsy. In a few days, she also developed left abducens nerve palsy and chemosis, exophthalmos, and proptosis of both eyes. There was pulsation over the left eye. Intraocular pressure was found to be elevated bilaterally. Selective carotid angiograms showed the presence of bilateral dural arteriovenous malformations (AVM) supplied by the external carotid arteries. Two months after the embolisation of the AVMs, eye movements improved. Repeat angiograms showed the absence of flow into the previously embolised AVMs. PMID- 11486557 TI - Symptomatic palatal myoclonus: an unusual cause of respiratory difficulty. AB - A 67-year-old man presented with dysphagia and difficulty breathing. Physical examination revealed palatal myoclonus. In this patient, the respiratory difficulty was caused by the fragmentation of breathing. Electromyographic examination of the cricothyroid muscle demonstrated rhythmic myoclonic jerks. Magnetic Resonance Imaging (MRI) yielded a pontine midline and right sided tegmental infarct. The patient responded to sodium valproate. PMID- 11486558 TI - Meningeal inflammatory pseudotumour: a case report. AB - We report the case of a meningeal inflammatory pseudotumour occurring in a 23 year-old male presenting with focal seizures and headaches. Brain imaging techniques showed a 3.5 cm left parietal meningeal tumour. Histology of the surgical specimen showed a dense lymphoid infiltrate permeating the dura mater and leptomeninges, consisting of a predominant polyclonal B cell population as confirmed by immunophenotyping and genotyping. Cultures of serum, CSF, and surgical specimen were negative and there was no serological evidence of a systemic dysimmune disease. The postoperative course was complicated by an episode of brain oedema resolving under steroid therapy. The patient, free from all medication, is asymptomatic at 3 years of follow-up. We discuss previously published cases and the nosology of intracranial inflammatory pseudotumours. PMID- 11486559 TI - Axillary injection of botulinum A toxin in a patient with muscle cramps associated with severe axillary hyperhidrosis. AB - Muscle cramps may be caused by fluid and salt loss induced by diffuse or focal hyperhidrosis. Recent reports have described the efficacy of botulinum, toxin in the treatment of primary focal hyperhidrosis. Botulinum toxin inhibits sweating by blocking exocytosis of acetylcholine from presynaptic cholinergic nerve terminals. We report the case of a patient who complained of frequent muscle cramps associated with unusually severe axillary hyperhidrosis. We used botulinum toxin to treat the excessive focal sweating presuming that it would also reduce the muscle cramps. A total dose of 200 MU of botulinum A toxin (Dysport) per axilla markedly reduced sweating and cramps. The beneficial effect started four days after the injection and it was still present five months later. Treatment was repeated in the sixth month with analogous results. No side-effects were observed and no compensatory sweating occurred. PMID- 11486560 TI - Cyclosporine-A induced neurotoxicity after renal transplantation. AB - Cyclosporine-A is a highly potent immunosuppressive agent for solid organ transplantation, but has many side effects including nephrotoxicity, hypertension, gum hyperplasia, hepatotoxicity, and neurotoxicity. Neurotoxicity is a less known toxic effect. The pathogenesis of this effect is unclear. However, it has been postulated that hypomagnesemia, hypocholesterolemia, corticosteroids, and/or neurotoxic substances can induce this syndrome. Also, it has been suggested that the endothelial damage caused by Cyclosporine-A may contribute to neuropeptide-mediated ischemia in the brain and lead to the development of neurological symptoms. In this report, we present a case with reversible neurologic deficits whose symptoms returned to normal after the cessation of cyclosporine-A. PMID- 11486561 TI - Subacute sclerosing panencephalitis in a vaccinated, internationally adopted child. PMID- 11486562 TI - The many faces of human prion diseases in Belgium and the world. AB - Prion diseases are rare neurodegenerative disorders that always lead to death and that can be transmissible under certain conditions. Although sporadic Creutzfeldt Jakob's disease (CJD) is the best known human variant of these transmissible spongiform encephalopathies with an incidence of about 1 in 106 inhabitants, several other types of human prion disease have been described (e.g. Familial CJD, Gerstmann-Straussler-Scheinker syndrome, Fatal Familial Insomnia,...). In 1996, a variant of CJD has been linked to the epidemic of bovine spongiform encephalopathy (BSE). Therefore, vigilance concerning prion diseases was increased throughout the whole of Europe. In Belgium, a comprehensive, nation wide study has been conducted both retrospectively (1960-1997) and prospectively (1998-...) to identify prion disease patients. In 1998, a surveillance system has also been created to monitor the incidence of CJD and other prion diseases. Using data from both studies and the surveillance program, the occurrence and phenotype of all types of prion diseases in Belgium was investigated. The sporadic type of CJD was identified in 116 patients, while 4 suffered from a hereditary form. In our series, we could find no evidence for variant or iatrogenic CJD, neither for the more rare types of prion diseases. PMID- 11486563 TI - Intravenous rtPA thrombolysis in acute ischemic stroke. AB - Early intravenous thrombolysis within the first three hours has been considered in the United States as the first proven treatment in acute ischemic stroke. However, not all patients will respond to this therapy which is also associated with a risk of symptomatic, including fatal, intracranial hemorrhage. This overview addresses the issue of efficacy and safety of intravenous alteplase (tPA) in acute cerebral ischemia. The rationale for thrombolytic therapy and its limits are described. The controlled studies show that intravenous tPA is effective and safe when given under restrictive conditions within 3 hours after stroke onset, but the data for a larger therapeutic window between 3 and 6 hours remain controversial. The expected functional improvement and the risk of intracranial hemorrhage greatly depend on selective clinical and imaging criteria. For this purpose, MRI, using the diffusion- and perfusion-weighted sequences combined with MR- angiography, should be preferred to CT scan in the next future. Applicability of tPA thrombolysis in current neurological practice in Belgium is discussed. Before its generalization, this therapy should be restricted to specialized stroke centers and all treated patients should be recorded in a central data bank to guarantee continued surveillance. PMID- 11486564 TI - Cerebellar neurocognition: a new avenue. AB - The functional role of the cerebellum is traditionally defined as a mere coordinator of automatic and somatic motor functions. This view determined most of the clinical and experimental investigations of the functions of the cerebellum and overshadowed many attempts to expand its role to non-motor functions. However, during the past decades the collaboration across contemporary neuroscience disciplines has brought about a substantial modification of traditional thinking about the cerebellum. Converging evidence from different neuroscientific angles established the view that the human cerebellum participates in a much wider range of functions than conventionally accepted. Studies have brought to the fore that the cerebellum also modulates neurocognitive functions of at least these parts of the brain to which it is reciprocally connected. In less than two decades, the concept of 'cerebellar neurocognition' evolved from a mere afterthought to an exciting new multifaced area of contemporary neuroscientific investigations. Within this area, one of the major avenues of current research is the role of the cerebellum in non-motor language processing. This paper reviews the recently acknowledged role of the cerebellum in neurocognition and linguistic processes. PMID- 11486565 TI - Surgical management of ulcerative colitis. AB - As long as our understanding of ulcerative colitis is too limited to allow a more specific, disease-targeted treatment, surgery will play an important role in the management of these patients. Careful interdisciplinary evaluation and counselling of patients with ulcerative colitis will permit to achieve the goals on an individual basis with maximum safety. Restorative proctocolectomy with an IPAA has evolved as the procedure of choice among four basic surgical options because it appears to be safe and carries a low mortality. Although the associated morbidity is not negligible, functional results are generally good and patient satisfaction is high. PMID- 11486566 TI - Treatment of primary and secondary liver malignancy. AB - The management of hepatic malignancy is one of the most controversial areas in medicine. It is a continuously evolving field which requires a multimodal approach and the inclusion of a medical oncologist, a hepatobiliary surgeon, a radiotherapist, an interventional radiologist and in some cases a transplant surgeon. In addition, the recent development of novel approaches such as neoadjuvant tumour therapy, cryosurgery, thermal ablative techniques as well as biological and immunological manipulation of malignant cells has added to the complexity of this field. Today, the availability of such innovative modalities, in the absence of an extrahepatic localisation of disease, allows the potential to cure many large primary and secondary hepatic tumours. However, the appropriate use of these various treatment modalities should be limited to centers with experience in treating patients with advanced disease where innovative study protocols are available. Experience of the surgeon with difficult hepatectomies is of paramount importance. Currently, mortality following major hepatectomies in non-cirrhotic patients is below 5% in centers of reference. For example, in the series from Duke University Medical Center there were no fatalities in more than 150 liver resections in patients with benign diseases and a 2% mortality in more than 300 patients with malignant diseases. Mortality, however, reached 5% in cirrhotic patients undergoing a resection involving more than 2 segments. This review will attempt to focus on an overview of the diagnosis and treatment of a selection of more common hepatic malignancies. PMID- 11486567 TI - Severe pulmonary hypertension: data from the Swiss Registry. AB - BACKGROUND: Severe pulmonary hypertension (PH) is a rare disease with a dismal prognosis if untreated. Progress in diagnosis and in the development of effective therapeutic options has created new interest in this pathology. There are, however, only limited data on the prevalence of severe PH unrelated to chronic left ventricular failure or COPD, on the associated conditions and on the parameters with a prognostic impact. With the aid of a retrospective registry we have collected data from 5 centres in Switzerland and attempted to answer the above questions. METHODS: Data on patients with PH from 4 university facilities (Zurich, Basle, Geneva and Lausanne) and one well-defined geographical area (Ticino) were retrospectively collected and analysed up to December 1999. Clinical and haemodynamic parameters and associated diseases were noted. We were also interested in the age distribution of the patients and the year of diagnosis of PH. RESULTS: We found 106 patients with severe PH (43 men, 63 women, median age 43 years); 79% were in NYHA class III or IV. There was a steep rise in diagnosis of PH after 1995. In 74% PH was either primary or associated with collagen vascular disease or thromboembolic disease. By the end of the observation period 30% of the patients had died. The best distinguishing parameters between surviving patients and those who eventually died were the 6 minute walking test (363 vs. 235 metres, p = 0.002), the NYHA class (II vs III/IV, p = 0.015), and mixed venous saturation (66.5 vs. 57.9%, p = 0.006). Therapy consisted of calcium antagonists in 18% and of (inhaled) prostanoids, chiefly iloprost, in 33%. Seven patients underwent lung transplantation. CONCLUSIONS: We conclude that PH is diagnosed more often as diagnostic and therapeutic options improve; that primary forms, and those associated with collagen vascular disease and with chronic venous thromboembolism, make up three quarters of the aetiologies; and that the 6-minute walking test, the functional class and mixed venous saturation are the best prognostic parameters. PMID- 11486568 TI - Sympathetic stimulation using the cold pressor test increases coronary collateral flow. AB - BACKGROUND: Little is known about the vasomotor function of human coronary collateral vessels. The purpose of this study was to examine collateral flow under a strong sympathetic stimulus (cold pressor test, CPT). METHODS: In 30 patients (62 +/- 12 years) with coronary artery disease, two subsequent coronary artery occlusions were performed with random CPT during one of them. Two minutes before and during the 1 minute-occlusion, the patient's hand was immerged in ice water. For the calculation of a perfusion pressure-independent collateral flow index (CFI), the aortic (Pao), the central venous (CVP) and the coronary wedge pressure (Poccl) were measured: CFI = (Poccl - CVP)/(Pao - CVP). RESULTS: CPT lead to an increase in Pao from 98 +/- 14 to 105 +/- 15 mm Hg (p = 0.002). Without and with CPT, CFI increased during occlusion from 14% +/- 10% to 16% +/- 10% (p = 0.03) and from 17% +/- 9% to 19% +/- 9% (p = 0.006), respectively, relative to normal flow. During CPT, CFI was significantly higher at the beginning as well as at the end of the occlusion compared to identical instants without CPT. CFI at the end of the control occlusion did not differ significantly from the CFI at the beginning of occlusion with CPT. CONCLUSIONS: During balloon occlusion, collateral flow increased due to collateral recruitment independent of external sympathetic stimulation. Sympathetic stimulation using CPT additionally augmented collateral flow. The collateral-flow-increasing effect of CPT is comparable to the recruitment effect of the occlusion itself. This may reflect a coronary collateral vasodilation mediated by the sympathetic nervous system. PMID- 11486569 TI - Cryptosporidiosis and travelers. PMID- 11486570 TI - Spinal epidural lipomatosis. PMID- 11486571 TI - [Atherothrombosis and inflammation]. AB - Alterations of coagulation, fibrinolysis, platelets and low grade inflammation are causal pathophysiological factors in atherosclerosis. Considerable activation of several involved pathways occurs during the acute progression of atherosclerotic lesions, which is characterized by an occluding thrombus, and local and systemic inflammatory reactions as in patients with acute coronary syndromes. These patients become clinically compromised due to the reduction in coronary flow. Furthermore, a frequent occurrence of non-occluding thrombi may be assumed as a progression factor in atherosclerotic diseases. Both the extrinsic and the intrinsic pathway of coagulation are involved, resulting in a hypercoagulative state. Furthermore, an inflammatory acute phase reaction occurs in addition to the activation of several other inflammatory pathways in patients with unstable angina pectoris or acute myocardial infarction. Exposure of tissue factor by the ruptured plaque together with a systemic hypercoagulative state, local and systemic inflammation as well as stimulated platelets and endothelial dysfunction are involved in the pathophysiology of acute coronary syndromes. In the following paper the current knowledge on activation of these pathways and on the various complex interactions is discussed. PMID- 11486572 TI - [Current status of endocarditis prevention]. AB - There are currently no randomized and carefully controlled human trials to definitively prove that endocarditis prophylaxis is efficient. Furthermore, most cases of endocarditis are not attributable to a medical procedure. Thus, even with a high level of application of endocarditis prophylaxis only a minority of cases could be prevented. Endocarditis is a rare disease. On the other hand, its morbidity is increasing! In addition, infective endocarditis remains still a major medical concern because of its mortality between 5% and 76%. In addition, in up to 40% of all patients suffering from endocarditis one or more heart valves have to be replaced in the following 5 to 8 years. Without treatment endocarditis has a lethality of 100%. Therefore, there is worldwide agreement that endocarditis prophylaxis is necessary. Combining the recommendations of the German and the American Heart Association, as well as the results of the European consensus conferences, with newer insights into the pathophysiology of endocarditis the following aspects are elucidated: depending on their risk of endocarditis patients are allocated into 3 groups. In the first group there are patients with prosthetic cardiac valves, patients who suffered from previous endocarditis and patients with complex cyanotic congenital heart disease and surgically constructed shunts or conduits of the aorta and/or pulmonary circulation. In these high-risk patients the prophylactic regimen for dental, oral, respiratory tract procedures is oral amoxycillin. In genitourinary and gastrointestinal procedures ampicillin and gentamicin i.v. is recommended. In patients with mostly congenital cardiac malformations, acquired valvular dysfunction, hypertrophic obstructive cardiomyopathy and mitral valve prolapse or thickened leaflets and valvular regurgitation oral amoxycillin is recommended for all medical procedures (second group). The third group consists of patients with isolated secundum atrial defect, previous coronary bypass graft surgery, patients with cardiac pacemakers or defibrillators. In this patient cohort the individual risk of endocarditis is not higher than in the general population. Therefore, endocardits prophylaxis is not recommended. PMID- 11486573 TI - [Current prescription practice of CSE inhibitors at clinic discharge after acute myocardial infarct]. AB - We investigated the use of statins in clinical practice in patients with acute myocardial infarction in Germany in 17,732 consecutively included patients of the registries MIR-1 and MITRA-1. A clinical follow-up has been performed in the MITRA-1 study after a mean period of 18 months. In total 30% of all patients with acute myocardial infarction received statins at discharge. From 1994 to 1998 the use of statins increased from 6% to 44%; however in 1998 still less than half of the patients with acute myocardial infarction received statins at discharge. In a logistic regression model, concomittant diseases as renal failure (OR 0.7), heart failure (OR 0.7) and diabetes mellitus (OR 0.9) were associated with a lower use of statins. Age > 70 years (OR 0.5) was also associated with a lower use of statins at hospital discharge. Patients with statins at discharge had a lower long-term mortality of 5.8% versus 12.9% in patients without statins. After adjustment to age and comorbidity, use of statins at discharge was associated with a borderline significant reduction of long-term mortality (multivariate OR 0.7, 95% CI 0.4-1.0). In a subgroup analysis of therapeutic benefit, measured by the "number needed to treat" (NNT), the number of patients to treat with statins to save one life, patients with cardiovascular risk factors, as heart failure (NNT 7.5), diabetes mellitus (NNT 7.8) and age > 70 years (NNT 13.8) had a larger therapeutic benefit as patients without these risk factors (NNT 345). However, these high-risk patients received less often statins than patients without risk factors (use of statins 11.8% versus 19.8%). PMID- 11486574 TI - Troponin T elevation after successful directional atherectomy. AB - BACKGROUND: The incidence and correlates of postinterventional cardiac troponin T (cTnT) elevation have not been evaluated in patients with successful directional atherectomy (DCA). METHODS: Total creatine kinase (CK) activity, CK-MB mass concentrations and cTnT levels were measured in 36 patients before, and serially 4, 24, 48 and 72 hours after successful DCA. Patients were followed for death, Q wave infarction, need for emergency bypass surgery, periprocedural vascular complications (transient in-lab vessel closure, side branch compromise, large dissection), and non-Q-wave infarction on ECG. Patients were followed for clinical outcomes and angiographic restenosis for 6 months. Vascular correlates were studied by coronary angiography and intravascular ultrasound before, immediately after and 4 hours after elective and successful DCA. RESULTS: 25 patients (69.4%) had elevated levels of cTnT, whereas CK-MB mass > or = 6 ng/ml and mild elevations of total CK activity were present in only 36.1 and 5.6%, respectively. Elevated cTnT was related to vascular complications in 44% of cases. Inapparent microembolization of platelets or plaque debris was considered responsible for most of the remaining micronecroses. During 6 month follow-up rates of clinical restenosis (44 vs 9%, p = 0.059) tended to be higher in patients with as compared to patients without cTnT release after DCA. CONCLUSION: Successful DCA is associated with postprocedural elevations of cTnT which relate to minor vascular complications and presumably microembolization of platelets or plaque debris. The superior diagnostic performance of cTnT compared to CK-MB mass may relate to more sensitive identification of microinfarction. Further studies are mandatory to confirm the association between elevation of cTnT and risk of restenosis and TVR. PMID- 11486575 TI - [Cardiac troponin I after heart surgery corrective operation in infancy and childhood]. AB - BACKGROUND: Perioperative myocardial damage is an important determinant for postoperative cardiac function and recovery. Cardiac troponin I (cTNI) is a specific marker for myocardial damage. The aim of our study was to evaluate pre- and postoperative cTNI levels, the pattern of elevation in the first four postoperative days and the prognostic value after pediatric cardiac operation. METHODS: Cardiac troponin I levels were measured in 115 children mean age 36 +/- 45 months (range 4 days to 189 months) undergoing elective operation of a congenital heart defect. Routine measurements were made preoperatively, immediately after cardiopulmonary bypass and serially 8, 18, 42, 90, 138 hours thereafter. Data from 13 patients undergoing surgery without cardiopulmonary bypass served as controls. Postoperative cTNI levels were correlated with intra- and postoperative parameters (such as duration of aortic crossclamping, cardiopulmonary bypass time and need for postoperative inotropic support). RESULTS: All preoperative cTNI levels were in the normal range. Postoperatively, the highest median cTNI levels were found in patients after repair of tetralogy of Fallot (TOF), atrioventricular septal defect (AVSD) and implantation of a homo or xenograft. Postoperative cTNI levels correlated significantly with duration of cardiopulmonary bypass and aortic crossclamping, operative approach (ventriculotomy versus atriotomy) and inotropic support (p < 0.0001). Peak cTNI levels were found immediately after surgery in 77.4% of our patients, 8 hours postoperative in 13.9% and at 18 hours after the surgery in 5.2% of the patients. In three children cTNI continued to increase; a secondary increase was found in one patient. Two of these children died, two had a prolonged postoperative recovery. CONCLUSION: The postoperative level of cardiac troponin I could be used as a marker of perioperative myocardial injury caused by ischemia and operative trauma. Peak levels usually could be obtained immediately after surgery, but a further increase of cTNI during the following 18 hours may occur and is not necessarily related to impaired recovery. However still increasing cTNI levels after 18 hours postoperatively and a secondary increase as well may be used as indicators of poor outcome. PMID- 11486576 TI - [A new combined surgical interventional method for univentricular palliation of congenital heart defects]. AB - Total cavo-pulmonary anastomosis is frequently performed to palliate patients with a broad variety of congenital heart defects with functionally univentricular hearts precluding biventricular circulation. In patients with risk factors for primary repair a stepwise approach is frequently chosen with initial creation of an aorto-pulmonary shunt followed by a Glenn anastomosis or hemifontan procedure. Finally a total cavo-pulmonary connection is completed surgically. The aim of this feasibility study was to develop a combined surgical-interventional approach for creation of a total cavo-pulmonary anastomosis which reduces the number of surgical interventions, precludes extracorporeal circulation and intracardiac sutures, putting the patients at risk for subsequent rhythm disturbances. METHODS: A Glenn anastomosis was created in 10 sheep without cardio-pulmonary bypass. The superior vena cava was banded superior to the cavo-atrial junction leaving a minimal lumen for subsequent interventional balloon dilatation and implantation of a stent graft. A 15 mm Gore-Tex tube was sutured around the inferior vena cava intrapericardially to provide resistance for subsequent interventional stent implantation. Total cavo-pulmonary anastomosis was completed interventionally by dilating the banded vena cava superior and connection of the inferior vena cava to the superior vena cava by implanting an Aneurx covered stent graft. RESULTS: All animals survived the combined surgical-interventional procedure. Stent deployment was possible without causing obstruction to the hepatic veins. Neither stent dislocation nor rhythm disturbances were encountered. CONCLUSIONS: Total cavo-pulmonary anastomosis can be achieved without intracardiac sutures and cardio-pulmonary bypass by use of a combined surgical-interventional approach using the Aneurx covered stent graft. PMID- 11486577 TI - [Safety and efficacy of LK565--a new polymer ultrasound contrast medium]. AB - We tested a newly developed ultrasound contrast agent (LK565) from poly-aspartic acid (PAA; particle size 3 microns; particle content: air) in 15 healthy male probands (20-38 years) in doses of 10, 30 and 100 mg intravenously. One day and immediately before the study a routine laboratory test, an ECG and an EEG were performed. The EEG was continued through the complete time period that the ultrasound contrast lasted, i.e., up to one hour after the injection. All probands were followed clinically for 24 hours when the routine laboratory and the ECG were repeated. All subjects tolerated the contrast agent well. There were no changes in either the EEG or in the ECGs performed throughout the study. There were no significant laboratory changes except for a small and transient increase in the neutrophil count in five probands receiving the highest dose. All injections with 10 mg led to a significant improvement in the color Doppler signal. All injections with 30 and 100 mg led to a very strong echo contrast lasting for 5 to 12 minutes in the harmonic B-mode. Using the latter, fragments of intramyocardial coronaries could be visualized. The tested ultrasound polymer contrast agent was safe, well tolerated and efficient in this acute study. PMID- 11486579 TI - [Circular hemorrhagic subendocardial necrosis after heart arrest and resuscitation]. AB - A 19 year old boy, previously healthy suffered a cardiac arrest by getting up. Ten minutes afterwards the general practitioner commenced cardiopulmonary resuscitation, 15 minutes later ventricular fibrillation developed and the emergency physician carried out intubation and repeated defibrillation. During the next few hours a stabilization of the circulatory system was achieved. Five days after the cardiac arrest brain death occurred. The postmortem findings on the heart (only a heart section was performed) showed extensive circular hemorrhagic subendocardial necrosis with initial organization of the left ventricular wall and the septum with a well-preserved subendocardial area. In the right ventricular wall only a few small areas of organization were observed. All lesions were consistent with the cardiac arrest suffered 5 days previously. The morphological changes differ from those of a usual hemorrhagic infarction and of the sequences of a cardiopulmonary resuscitation. The circular subendocardial necroses occur after a cardiac arrest which exceeds the resuscitation time of the heart. They do not respect the area of coronary distribution and their hemorrhagic component develops after successful reanimation within the necrotic myocardium. PMID- 11486578 TI - [Left ventricular diastolic function in pregnancy in patients with arterial hypertension. A prospective study with M-mode echocardiography and Doppler echocardiography]. AB - INTRODUCTION: During pregnancy eminent cardiovascular changes occur. The aim of the following study was to investigate the course of hemodynamic parameters under increased volume load during pregnancy in women suffering from mild arterial hypertension. METHODS: Altogether 47 women (age: 25 +/- 4 years) with mild arterial hypertension detected during pregnancy underwent echocardiography at the 9th, 24th and 33rd week of gestation. Furthermore echocardiography was performed postpartum at weeks 1 and 8. The control group comprised 45 healthy pregnant women. In all patients left ventricular muscle mass index and systolic shortening fraction were measured. The following Doppler echocardiographic parameters were ascertained: peak early diastolic and peak late diastolic flow, VE/VA ratio, acceleration time, deceleration time and isovolumetric relaxation time. RESULTS: During pregnancy all patients had an increase of left ventricular muscle mass index and a decrease of fractional shortening. All patients developed a relevant diastolic dysfunction. While the control group developed signs of disturbed relaxation as reduction of peak early diastolic flow (0.89 +/- 0.07 versus 0.82 +/- 0.08 m/s*), VE/VA ratio and an increase of isovolumetric relaxation time (72 +/- 12 versus 123 +/- 7*) at the 33rd week of gestation (* p < 0.01), all pregnant women with mild arterial hypertension developed a diastolic dysfunction with signs of delayed relaxation already at the beginning of gestation. 26 pregnant women with arterial hypertension developed a restrictive diastolic filling pattern at 24 weeks of gestation. The other 21 pregnant women only showed restriction for a short time at the end of gestation. In healthy pregnant women, volume load results in a reversible physiologic left ventricular hypertrophia, a significant alteration of diastolic left ventricular function in terms of a disturbed relaxation pattern and a temporary decrease of systolic function. In comparison hypertensive pregnant women show a delayed relaxation at the beginning of pregnancy and 50% developed early signs of restrictive cardiomyopathy. These changes may predispose to critical complications during pregnancy. PMID- 11486580 TI - [Recommendations for qualification in cardiovascular magnetic resonance tomography]. PMID- 11486581 TI - Coronary interventions in the elderly--the German ALKK Study Group experience. PMID- 11486582 TI - [Has tongue base reduction with radiofrequency energy in sleep apnea syndrome been adequately evaluated?]. PMID- 11486583 TI - [Robotics. A new dimension in otorhinolaryngology?]. AB - Robots have entered into many aspects of human life, including operative medicine. However, current medical robots have nothing in common with anthropomorphic robots as known in science fiction novels. We distinguish manipulators, working on a master-slave principle, from robots. Robots can be defined as "automatically controlled multitask manipulators, which are freely programmable in three or more axes." The success of robots is based on their precision, lack of fatigue, and speed of action. Potential fields of application for manipulators lie in endonasal surgery and for robots in lateral skull base surgery, including mastoidectomy and drilling a cavity for implantable hearing systems and cochlear implants. We performed a number of experiments at the department of anatomy with respect to robotic lateral skull base surgery. This paper reviews the current use of manipulators and robots in operative medicine and their potential applications in otorhinolaryngology. PMID- 11486584 TI - [Applications for a robot in the lateral skull base. Evaluation of robot-assisted mastoidectomy in an anatomic specimen]. AB - In the past decade, a great variety of robot systems have been applied in numerous areas of life. In the public health system, robots are increasingly used in the operating theater. The potential for reproducibility and predictability was one of the main arguments for the use of robots in orthopedic treatment of bones, especially the implantation of a cementless total hip replacement. In otorhinolaryngological surgery (ENT), different hearing aids were developed: the cochlea implant for the deaf or the totally implantable hearing aid for the hearing impaired. Their site of implantation is localized in the lateral skull base. Removal of the bone mass with the reamer requires both great precision and considerable physical effort on the part of the surgeon, which does not result in an ergonomic operating technique. The following project describes the evaluation of processing parameters for a robot-assisted mastoidectomy to expose an implantation bed. The goal was to establish different parameters for robot controlled reaming in the calotte or mastoid. In addition, several parameters were tested for their influence on surface structure, procedure reliability, and quality as well as the ability of the Mayfield clip to stabilize the head during the operation. PMID- 11486585 TI - [The NEN (Nicolet Electronic Naviation System) ENT navigation system. Initial clinical application]. AB - BACKGROUND: Intraoperative computer navigation will soon play an important role in procedures performed on the anterior and lateral skull base. Electromagnetic systems compared to optical systems offer some advantages such as small hardware dimension and free unimpaired mobility of the surgeon. PATIENTS AND METHODS: The NEN navigational system (Nicolet Electromagnetic Navigation System) was used in 22 patients who underwent microscopic/endoscopic surgery for polypoid rhinosinusitis. It is the prototype of a new system, which localizes the surgical probe via the measurement of pulsed electromagnetic DC fields. This system was modified for the needs of head and neck surgery. Preoperative imaging data consisted of an axially oriented spiral computed tomography (CT) resulting in a slice thickness of 1 mm with a radiation dose slightly higher than a routine coronary CT scan of the paranasal sinuses. RESULTS: The data acquired during clinical application was used to optimize navigation accuracy. The number and positions of the markers were gradually modified and improved. Six markers including the volume of the paranasal sinus system (three non-coplanar frontal markers, one on each of the mastoid processes and one median marker on parietooccipital junction) offered the best navigation precision. Additionally, all surgical instruments measured as ferromagnetic, i.e., nasal specula, were replaced by titanium instruments. An adapter affixed to the maxilla-designed to hold a second sensor used to track the patient's head was developed. With these modifications, it was possible to improve navigation accuracy to 1.5 mm in the plane of the sphenoid bone while retaining free head movement of the patient. CONCLUSION: For routine procedures such as microscopic/endoscopic endonasal sinus surgery, the systems have to be cost effective and easy to operate. PMID- 11486587 TI - [Permeability of the round window membrane for prednisolone-21-hydrogen succinate. Prednisolone content of the perilymph after local administration vs. systemic injection]. AB - BACKGROUND AND OBJECTIVE: Prednisolone is the drug of first choice for the treatment of cochleovestibular disorders, such as sudden hearing loss. Because of the known side effects, the efficient drug levels to be achieved within inner ear fluids are limited by intravenous administration. The aim of the study was to determine the concentration in the perilymph of prednisolone-21-hydrogen succinate applied into the round window niche in comparison to the concentration after intraperitoneal application. METHODS: Application of prednisolone-21 hydrogen succinate (5 mg in 0.1 ml) on the round window membrane was performed after sedation under microscopic view directly into the round window niche of the guinea pig. In order to compare the results, perilymph samples after systemic application of 60 mg/kg body weight prednisolone were used. The time between application and taking specimens of perilymph from the cochlea varied. Specimens of perilymph were obtained after 15, 20, 80, 180, 330, and 960 min (10 specimens in each group, n = 60) by dissecting the cochlea and opening the apex cochleae. Levels of prednisolone-21-hydrogen succinate in perilymph were measured by isocratic high-pressure liquid chromatography (HPLC). RESULTS: The highest levels of prednisolone-21-hydrogen succinate were found after 180 min: 952.3 mg/l (95% confidence interval: 382.7). After 960 min the level was 18.72 mg/l (95% confidence interval: 16.9). In the group with systemic application, the levels measured were below 14.71 mg/l (95% confidence interval: 7.05). CONCLUSION: The results demonstrate that high levels of prednisolone-21-hydrogen succinate in perilymph are achievable by local application of a single dose into the round window niche. After application of 5 mg, the levels of prednisolone are measurable up to 16 h. PMID- 11486586 TI - [Tongue base reduction with radiofrequency energy in sleep apnea]. AB - BACKGROUND AND OBJECTIVE: Tongue base reduction with temperature-controlled radiofrequency for the treatment of obstructive sleep apnea syndrome is a minimally invasive technique. Repeated application leads to a progressive shrinking of the tissue. PATIENTS AND METHODS: In our study, we summarize the experiences gained from 100 tongue base reductions and compare them with the pilot study that was recently published. An intensified treatment scheme was used with higher amounts of energy applied per treatment session. Visual analogue scales were used for the assessment of postoperative pain and functional parameters. Regular follow-up visits were scheduled to evaluate postoperative complications. RESULTS: Postoperative pain was mostly mild or moderate. Paraoperative complications were not observed. The overall rate for postoperative complications was 8%, with 2% mild and 5% moderate complications. One severe complication--a tongue base abscedation--was observed. Using para- and postoperative antibiotic prophylaxis reduced the rate of complications. Functional parameters such as taste or swallowing were not affected. CONCLUSIONS: Our results underline the safety of the procedure and demonstrate the minimal para- and postoperative morbidity. The increased amount of energy applied per session has not led to an increase in postoperative morbidity. PMID- 11486588 TI - [Is recurrent loss of low frequency tone perception--without vertigo--a precursor of Meniere disease?]. AB - PURPOSE: Recurrent low-frequency sensory hearing loss is one characteristic sign of Meniere's disease. It is therefore often assumed to be a prodromal sign of Meniere's disease. METHODS: We report on 81 patients with recurrent low-frequency sensory hearing loss who did not previously suffer from vertigo. All patients underwent a follow-up examination after at least 1 year up to more than 10 years after the onset of the disease [average: 64.65 months (SD 56.33)]. CONCLUSIONS: Only 3.7% of these patients developed the typical signs of Meniere's disease with labyrinthine vertigo, whereas 25.9% suffered from recurrent hearing loss and an unspecific vertigo, which could be diagnosed and treated as psychogenic vertigo; 3.7% showed a typical benign positional vertigo. We can thus conclude that although almost every patient with Meniere's disease suffers from recurrent hearing loss, only a few patients with recurrent hearing loss will develop Meniere's disease. However, many patients with low-frequency sensory hearing loss develop anxiety and psychogenic dizziness in expectation of "imminent" Meniere's disease. Therefore, it is important to counsel the patients carefully that the probability of this occurring is not very high. PMID- 11486589 TI - [Botulinum toxin type A-induced "rebalancing" in bilateral vocal cord paralysis?]. AB - BACKGROUND: Upper airway obstruction due to bilateral vocal cord paralysis in an 80-year-old female patient was successfully relieved by injection of botulinum toxin A (BTA) into the laryngeal adductor muscles. The patient achieved satisfactory airway ventilation. Spirograms obtained preoperatively and postoperatively documented improved peak flow rates and 1-s forced expiratory volume values. Voice quality was breathy after the injection; however, neither aspiration nor dysphagia developed. Surprisingly, the maximum phonation time increased. PATIENTS AND METHODS: During a follow-up check 4 months later, the patient still reported less dyspnea although the vocal cords were closer together than initially after the injection. The decrease in dyspnea as reported by the patient lasted approximately 2 years. RESULTS: The improvement in breathing following injection of BTA can be interpreted as a paralysis or weakening of the laryngeal adductors. However, it remains unclear why the maximum phonation time increased. Comparable findings, i.e., improvement in overall laryngeal function, are described in the literature as BTA-mediated laryngeal rebalancing. PMID- 11486590 TI - [Adult multilocular rhabdomyoma as etiology of a space-occupying lesion in the area of the neck]. AB - Real rhabdomyomas belong to the rarest tumors in the human body. The origin of these benign neoplasms is in the cardiac or skeletal muscle, in the beginning, there are no significant symptoms and the tumor grows very slowly. Extracardiac rhabdomyomas seldom occur and are mostly located in the head and neck region. A multilocular occurrence represents a rarity. We describe a 77-year-old female patient with a multilocular adult rhabdomyoma in the parapharyngeal and paratracheal space. The tumor was accidentally found; the patient had not complained of any specific symptoms. After histological examination, complete resection was carried out. On discharge the patient showed no functional deficits. Long-term follow-up is necessary because local recurrences have been described. PMID- 11486591 TI - [Hypertrophy of the salivary glands in bulimia]. AB - We report a case of bulimia nervosa with bilateral swelling of the parotid and submandibular glands. It was the only symptom of the behavioral disorder. Sjogren's disease was excluded histologically. In our case, the parotomegaly as a symptom of sialadenosis was found at the same time as the eating disorder. MRI or ultrasound are important diagnostic tools to confirm the diagnosis. PMID- 11486592 TI - [Isolated uncertain tympanic membrane findings. Central site of tympanic membrane cholesteatomas]. PMID- 11486593 TI - [Flexible versus rigid laryngoscopy and stroboscopy. Differential findings in voice disorders]. PMID- 11486594 TI - [Conservative treatment of non-tumor laryngeal diseases with hoarseness as the leading symptom]. PMID- 11486595 TI - Medicare reform: a Democrat's view. PMID- 11486596 TI - Rescue of retinal function by macular translocation surgery in age-related macular degeneration and other diseases with subfoveal choroidal neovascularization. AB - Age-related macular degeneration (AMD) is the leading cause of legal blindness among the elderly. In AMD and some other macular diseases, subfoveal choroidal neovascularization (CNV) damages the underlying retinal pigment epithelium (RPE), and because retinal function is dependent on a healthy RPE, vision is markedly reduced by a subfoveal CNV. To treat such CNVs, macular translocation surgery has been performed to move the sensory retina from the damaged RPE to healthier RPE. At present, this surgery is the only possible treatment to improve the visual acuity of patients with subfoveal CNV. Macular translocation surgery involves the detachment of the entire retina from the RPE by a subretinal infusion of fluid and creating a 360 degrees circumferential retinotomy followed by the rotation of the retina. Severe postoperative complications such as recurrent retinal detachment have been reported in about 30% of the cases after macular translocation. To determine the efficacy of this surgery, it is necessary to demonstrate an improvement in macular and overall retinal function objectively as well as subjectively. To this end, we have assessed the changes in visual function by measuring the visual acuity subjectively, and the macular function objectively by focal macular ERGs (FERGs). We shall show that there is an improvement in the FERGs in most patients after retinal translocation surgery but the full-field ERGs were reduced by about 30%. Thus, macular translocation surgery with 360-degree retinotomy may be feasible for macular function, although some degree of peripheral retinal function is lost. PMID- 11486597 TI - Human herpesvirus 6 infection in transplantation. AB - Human herpesvirus 6 (HHV-6) is ubiquitous in the human population and causes exanthem subitum, a benign disease seen in infancy. The virus remains latent in the body after primary infection, and reactivates in immunocompromised patients. Infection occurs in nearly half of all bone marrow or solid organ transplant recipients 2-3 weeks following the procedure. It has been suggested that the viral infection and activation result in clinical symptoms including fever, skin rash, pneumonia, bone marrow suppression, encephalitis, and rejection. In order to control the viral infection, several studies investigating the route of viral transmission and diagnostic procedures have been carried out. PMID- 11486598 TI - [Phenomena of intermittent rhythmic photostimulation in neuronal plasticity]. PMID- 11486599 TI - Quantitative estimation of dietary energy deficiency and effects of its supplementation on protein nutritional status of nondiabetic uremic patients undergoing protein restricted dietary regimens. AB - In chronic renal failure (CRF) patients with a reduced protein intake, if the patients' energy intake could be estimated on the basis of biochemical data together with protein intake, it would be easier to provide them with adequate dietary treatment. Thus, from the relationship among the normalized protein catabolic rate (nPCR) and the intrinsic creatinine generation rate (%GCr) both calculated on the basis of 24-hr urine creatinine, as well as the daily dietary energy intake evaluated by a skilled nutritionist, we devised the following equation to estimate the amount of dietary energy deficiency (delta E) whose supplementation increases the %GCr of patients on protein-restricted dietary regimens to the target level (i.e., the dietary energy deficient amount). This was done by taking the %GCr of average nondiabetic hemodialysis patients of the same age and sex as a temporal target level: delta E = [31.22 - 1.97 (%GCr)0.6]/(nPCR)0.15. In order to examine the clinical usefulness of this equation, the daily dietary energy deficient amount calculated by the equation was supplemented with protein-free jelly. As a result, the %GCr increased from approximately three-fourths of the target level to the target level within 4 months. PMID- 11486600 TI - Spatial image of school environment in children with cleft lip and/or palate. AB - To evaluate the adaptability of children with cleft lip and/or palate to school, their mental images of the school environment were investigated in a semantic differential method survey using 23 pairs of adjectives. The following eight places on and off the school grounds were chosen as environmental factors; classroom, gymnasium, school nurse's office, teachers' office, playground, hallway, road leading to school and road leading back home. 50 children with cleft lip and/or palate (ages 10-11 years) were enrolled in this study. These children had a feeling of liberation in the teachers' office and school nurse's office. However, they did not have an established feeling of relaxation or affinity in the classroom, gymnasium, playground, hallway, or on the road to and from school, the very locales that were the main areas of their school activity. This suggested that self-expression and adaptation were difficult for these children. PMID- 11486601 TI - Body iron stores and iron restoration rate in Japanese patients with chronic hepatitis C as measured during therapeutic iron removal revealed neither increased body iron stores nor effects of C282Y and H63D mutations on iron indices. AB - Information on the level of iron stores in chronic hepatitis C is clinically important because its reduction is technically simple and therapeutically effective. This study was performed to measure the levels of iron stores from the total amounts of hemoglobin removed during iron reduction therapy. The C282Y and H63D mutations of HFE gene were analyzed in 94 patients. All of the patients were negative for C282Y mutation. One patient was homozygous, and 4 patients were heterozygous for H63D mutation. The body iron stores and iron restoration rate were measured in 59 patients in serial courses of iron reduction therapy. Mean values of body iron stores in the two groups with and without H63D mutation were 890 and 606 mg, while those of iron restoration rate were 1.85 and 1.52 mg/day, respectively. None of the indices of iron metabolism were different from the reference values measured similarly in healthy subjects, suggesting that the iron deposition in chronic hepatitis C is limited to the liver, probably due to changes in the iron distribution in tissues. PMID- 11486602 TI - Simple synthesis of condensed pyran containing compounds and their antimicrobial properties. AB - Reaction of various fused pyran compounds with formic acid was studied. Thus, refluxing 6-aminopyrano[2,3-c]pyrazole-5-carbonitriles 3 with formic acid afforded the corresponding 3-aryl-3-(5-hydroxy-3-methyl-1H-pyrazole-4 yl)propanoic acids 4. Whereas, reaction of formic acid with 2-amino-4H-1 benzopyran-3-carbonitriles 6 gave the corresponding quinoline-2,5(1H,6H)-diones 7. The study was also extended towards many spiro compounds possessing pyran residue. The antimicrobial properties of the prepared compounds was screened. PMID- 11486603 TI - Synthesis and antimicrobial activity of novel pyrazole, pyrazoline, pyrazolinone and pyrazolidinedione derivatives of benzimidazole. AB - Four novel series of pyrazolylbenzimidazole derivatives have been prepared, namely 2-[(1-substituted phenyl-3,5-dimethyl-4-pyrazolyl)methyl]benzimidazole 5a d 2-[(1-substituted phenyl-3-methyl-5-oxo-4,5-dihydro-4-pyrazolyl-4 yl)methyl]benzimidazoles 6a-d; 2-[(1-substituted phenyl-3,5-dioxopyrazolidin-4 yl)methyl]benzimidazoles 7a-d and 2-[(4-(1-phenyl-5-aryl-4,5-dihydro-3 pyrazolyl)phenylaminoacetyl]thio- methyl)-benzimidazoles 12a-e. The antimicrobial testing of the prepared compounds was performed using Escherichia Coli (NCTC 5933) as Gram-negative bacteria, Staphylococcus aureus (NCTC 4163) as gram positive bacteria and Candida albicans (NCTC 5310) as yeast like fungi. The most potent compound was the pyrazolone 6a which exhibits interesting antibacterial activity against the gram-negative bacteria E. coli. PMID- 11486604 TI - Synthesis of some pyridone derivatives. AB - Chlorination of 6-aryl-3-cyano-2-pyridone-4-carboxylic acid (1) afforded the corresponding acid chloride (2) and the 2-chloro derivative (3). Esterification of (2) gave the corresponding esters (4a; b). Hydrazinolysis of (4a) afforded the respective pyridazinone derivative (5). Treatment of 6-aryl-2-chloro-3-cyano-4 pyridine carboxylic acid (3) with acetyl hydrazine, gave the triazinopyridine derivative (6), while treatment of 3 with sodium azide in DMF afforded the tetrazinopyridine derivative (7). Treatment of the N-acetyl derivative (1b) with thiosemicarbazide and/or hydroxylamine hydrochloride, yielded the correspoding semicarbazone and oxine derivatives (8) and (10), respectively. The reaction of 6 aryl-3-cyano-1,2-dihydro-2-thioxo-4-pyridinecarboxylic acid (1c) with ethylchloro acetate and/or thiourea yielded the mercapto ester derivative (11) and the corresponding pyrido [2,3-d] pyrimidine thione derivative (12). Condensation reaction of (1d) with anthranilic acid, afforded the quinazolone derivative (11). The reactivity of 11 towards hydrazine hydrate was investigated. The structural assignment of the new derivatives were based on IR, 1H NMR and mass data. Some of the new compounds were screened, in vitro, for antimicrobial activity and the results were encouraging. PMID- 11486605 TI - Cytotoxic bicyclic diterpene from the brown alga Sargassum crispum. AB - Study of the brown alga Sargassum crispum collected from Red Sea resulted in the isolation of new diterpene with hydroazulene skeleton, Sargassinone (6), some fatty acids ethyl ester andsome fatty acids. The identification of the isolated metabolites was established mainly by spectral methods and chemical transformation of sargassinone (6) to its acetate (7). The two diterpens (6, 7) exhibited substantial cytotoxic activities, as indicated by their IC50 values at the dose of 10 micrograms/ml or less. PMID- 11486606 TI - Use of sephacryl high resolution (HR) in tetanus anatoxin purification. AB - The tetanus purified anatoxin is used in the preparation of the tetanus toxoid and multiple vaccines (dT, DT and DTP), all of them strictly following specifications established by the WHO with a minimum antigenic purity equal to 1,000 Lf/mgPN. Aiming to establish more sensitive and accurate methods for purification, samples from four different lots of tetanus anatoxin were submitted to gel filtration in twenty independent trials using the Sephacryl S-100 HR and S 200 HR resins. The Authors were careful to optimize their parameters of performance as to sample volume, elution and selectivity flow for tetanus anatoxin purification, allowing their use in industrial scale. The Sephacryl S 100 HR resin presented the best selectivity, that is, the best separation, allowing a greater linear-flow and, consequently, the best purity index. Satisfactory results were also achieved with the Sephacryl S-200 HR resin after optimization of chromatographic parameters for elution flow and volume of the sample applied. The good results of purification obtained, as well as the high chemical stability, have pointed out both the Sephacryl S-100 HR and S-200 HR resins as equally efficient for industrial production. PMID- 11486607 TI - Antioxidant activity of the microalga Chlorella vulgaris cultered on special conditions. AB - The chemical composition of Chlorella vulgaris indicates that it has a high nutritional value to a wide range of essential nutrients, such as vitamins, minerals and proteins. Moreover, it contains other compounds such as n-3 and n-6 polynsaturated fatty acids, provitamins and phenolic compounds. In addition, this alga can be produced in large-scale systems. The objective of the present study was to evaluate the antioxidant capacity of a Chlorella cultured on three differents temperatures (15 degrees C, 20 degrees C and 30 degrees C) in 3 Klux. Chlorella cultured samples were submitted to sequential extration using as solvents: ether, methanol and water. The antioxidant activity in the extracts was measured by b-carotene/linoleic acid system, at 50 degrees C and absorbances reading at 470 nm. One control with BHT, 100 ppm was used in this determination. The total phenolic compounds was determined with Folin-Ciocalteu reagent using the spectrophotometric measured at 780 nm with catechin as standard. The phenolic acid analysis were carried out using gas chromatograph equipped with a capillary column and flame ionization detector. Non conjugated and total phenolic acids were identified on the basis of the relative retention time of their derivatives compared with the standard phenolic acids. The methanolic extract from Chlorella cultured at 30 degrees C showed higher antioxidant activity (85%) quite similar of BHT (86%). By the Rancimat test (lipidic medium) two fractions from methanolic extracts showed too higher antioxidant activity with induction times > 37.50 h at 60 degrees C and 11.5 h at 100 degrees C. The total phenolic compounds were 24.95 mg in 1 g of dry alga matter from methanolic extract and five phenolic acids were identified. The phenolic compounds salicylic, trans cinnamic, synaptic, chlorogenic, chimic and caffeic acids found in the methanolic Chlorella extract may be responsible for its higher antioxidant activity. PMID- 11486608 TI - Initial studies on the administration route of prolactin. AB - The influence of administration route of prolactin on its biological availability in rats was determined. Prolactin was administered intraperitoneally, intramuscularly and intranasally (intranasal inhalation) in single doses of 2.5 mg/kg body weight. Within 5 hours after the administration, there were noticed evident differences in prolactin concentrations in blood dependening on the route of administration. The maximal prolactin concentration after its intraperitoneal administration was 1.5 times as high as the maximal concentration observed for intramuscular or intranasal administration. The area under the curves: concentration: intraperitoneal administration time and concentration: intramuscular administration time was 132.99 ng/cm3/h and 136.28 ng/cm3/h, respetively. The area under the curve (AUC0-&) for the intranasal administration of prolactin was not much different than for the intraperitoneal and intramuscular ones, and was 111.30 ng/cm3/h. PMID- 11486609 TI - Studies of the complexation of chloranilic acid with proguanil. AB - Studies on the complexation between chloranilic acid and proguanil has been carried out. The chloranilic acid acted as a pi-electron donor. The stoichiometric relationship in the complex was determined by employing the Job's method of continuous variation. The results indicate that the complex exhibits a purple colour and absorbs maximally at 520 nm. Spectrophotometric studies show that the method could be useful in the analysis of proguanil. PMID- 11486610 TI - Influence of surfactants additions on the absorption process of nifedipine in vitro. AB - The absorption process of nifedipine in aqueous NaCl solutions with additions of the following surfactants: egg lecithin, sodium lauryl sulphate and Tween-80 was studied in vitro. A flow-through apparatus containing a piece of small intestine (ileum) of a rat was used for the absorption rate measurements. The surfactants were added in two concentrations, i.e., below and above their critical micellar concentrations respectively. The small additions of the studied surfactants to the aqueous nifedipine solutions (i.e. in the concentrations below the critical micellar point) notably increased the absorption rate of nifedipine. However, when the mentioned surfactants were added in higher concentrations (i.e., above their critical micellar points), a important decrease of the absorption of nifedipine was observed. PMID- 11486611 TI - Effect of activated charcoal on isoniazid absorption in rabbits. AB - The effect of activated charcoal on gastrointestinal absorption of isoniazid (INH) was determined quantitatively in rabbits. The presence of activated charcoal resulted to about 40% reduction of INH bioavailability as was indicated by reduction in the AUC0-24 hour. The plasma half live (T1/2) and peak plasma concentration (Cmax) of the drug were reduced, while the apparent volume of distribution (Vd) and total clearance (Cl) of the drug were increased in the presence of activated charcoal. The results present an assessment of adsorptive capacity of activated charcoal for INH in vivo and as such could serve as a cheap, effective and readily available means of emergency treatment of INH poisoning. PMID- 11486612 TI - Prediction of in vivo bioavailability of six brands of ciprofloxacin film coated tablets using the concept dissolution efficiency (DE). AB - A comparative in vitro dissolution efficiencies (DE) of six commercial brands of ciprofloxacin tablets were evaluated in acetic acid and phosphate buffer (pH 7.4) and results obtained were used in ranking of their probable in vivo bioavailability. The dissolution efficiencies (DE) of the six brands varied widely in the two media. This was attributed to differences in solubility of the drug in the two media. The dissolution efficiencies of five out of the six brands (Citrovenot, Ciproxin, Cipoxin, Ciproflox, and Quflox), in 0.1 N acetic acid fell within 60-75% at 30 minutes, and therefore, could be considered bioequivalent. The dissolution efficiency of a brand, Cipro, fell below 40% in the same medium and at the same sampling time and it was considered to be most likely less bioavailable in vivo. There was absence of correlation between the hardness and disintegration time of the brands with their dissolution efficiencies. PMID- 11486613 TI - [Quality control changes in release controller system]. PMID- 11486614 TI - Polycyclic 4(3h)-quinazolinones survey of biological properties (Part II). AB - The authors present examples of polycyclic 4(3H)-quinazolinones of natural origin. Structures and syntheses of pharmacologically active (antibacterial, antifungal, antihypertensive and cardiovascular, antiasthmatic, anti-inflamatory, CNS-depresant, secretion of gastric acid inhibitors) compounds are described. PMID- 11486615 TI - Cholinergic mechanism in imipramine and morphine antinoception. AB - Antinociceptive effect of imipramine and morphine was studied in rats using tail flick method. Morphine and imipramine both increased the latency of tail flick response. Pretreatment with naloxone (0.5 mg kg-1) decreased the antinociceptive effect of morphine (1 mg kg-1) and imipramine (10 mg kg-1). In order to see if antinociceptive effect of morphine or imipramine involve cholinergic mechanisms, mecamylamine (1 mg kg-1) or atropine (1 mg kg-1) were administered 30 minutes before morphine (1 mg kg-1) or imipramine (10 mg kg-1) administration. Mecamylamine decreased the antinociceptive effect of morphine and imipramine (P < 0.001), whereas atropine had no effect. Thus, opiodergic and nicotinic systems appear to be involved in morphine and imipramine induced antinociception. PMID- 11486616 TI - Cis- and trans-platinum and palladium complexes: a comparative study review as antitumour agents. AB - A large body of novel platinum and palladium complexes, in both the cis- and trans-forms, with various donor ligands, e.g. beta-carboline alkaloids, pyrazoles, DMSO, ferrocenylphosphines,...... have been tested for their antitumour activity against number of fluid suspension (P388, L1210, K562, and Raji) and solid tumour (KB, T47D, SW948, HeLa, A549, L929, Hep-2, RD,...) cell lines. Remarkable cytotoxic effects against these cell lines were observed by some of these complexes. The preliminary results indicated that most of the trans palladium complexes showed a better activity than the cis-platinum isomers and superior activity than that of the cis-palladium isomers. More importantly they showed activities equal to (or superior than) those of cisplatin, carboplatin and oxaliplatin (the anti-cancer drugs) in vitro. Although these results are preliminary, however, encouraging since they are in a disagreement with the previous studies that cis-isomers are more active than trans-ones; the complexes which have not received the required attention from the vast number of researchers in this field. PMID- 11486617 TI - [100 years of the German Orthopedic Society: from DGOC to DOG and DGOT to DGOOC]. PMID- 11486618 TI - [Ablative and extremity salvage tumor surgery of the lower extremity--a 10 year comparison]. AB - AIM: The treatment of choice for local tumor control was amputation in the 1970's. Nowadays, limb salvage procedures have become the new standard, implicating that limb salvage surgery results in a better quality of life. This study attempts to prove this hypothesis. METHOD: In total, 102 patients who survived longer than ten years after tumor treatment of the lower extremities were investigated, of these, 71 patients underwent ablative procedures compared to 31 patients with limb salvage surgery. Operative revisions, education level, and occupational situation were evaluated in both groups. To analyze the outcome of every patient regarding functional results, quality of life, life contentment, and social parameters, the functional evaluation system of the Muskulo-Skeletal Tumor Society (MSTS), the Freiburger Life-Contentment-Questionnaire (FLZ) and the Quality of Life Questionnaire (QLQ-C-30) of the European Organization of Research and treatment of Cancer (EORTC) were used. RESULTS: Patients treated with a limb salvage procedure underwent more surgical revisions (p < 0.000). Educational level and occupational situation showed no difference in both groups. Functional results reached similar levels in both groups (74.6% vs. 73.8%). Life contentment and Quality of Life measurements showed good results in both groups. The FLZ questionnaire showed significantly better results for the ablative group in some items. CONCLUSION: The type of surgical local therapy of lower extremity tumors has no measurable effect on quality of life according to long-term follow-up in lower extremity tumors. In cases with a risk of inadequate margins when performing limb salvage surgery, an ablative procedure should be preferred. PMID- 11486619 TI - [Characteristics of the learning curve in total hip endoprosthesis exemplified by the BiContact prosthesis]. AB - AIM: Total hip arthroplasty is a common operation but there are few data describing the learning curve of this operation. The aim of the presented study is to prove and describe the learning curve in total hip arthroplasty. METHOD: Between 1991 and 1993, 168 patients were included in this prospective study, operated by three surgeons having different experiences in operating total hip arthroplasty. Perioperative complications and postoperative X-rays were analysed. Patients were re-examined clinically and radiologically and by a questionnaire 5 years after operation. RESULTS: Radiological complications of the first 84 operated patients were statistically significantly higher than those of the following 84 patients, except for minimal bone fractures and leg lengthening. The learning curve of all surgeons was finished after 20 operations independent of their experience. Clinical results were not influenced by the learning curve at follow-up. CONCLUSIONS: Regardless of the experience of the surgeon, the learning curve in total hip arthroplasty is finished after about 20 operations for each surgeon. Intensive preoperative planning and exchange of experience inside and outside every clinic will shorten the length of the learning curve. PMID- 11486620 TI - [Risk factors for dislocation of a cementless total hip endoprosthesis--a statistical analysis]. AB - AIM: This retrospective study aimed at the statistical identification of risk factors for dislocation for a certain cementless hip endoprothesis. MATERIAL AND METHOD: At our department 2605 primary total hip arthroplasties were performed between 1987 and 1997. In 40 patients (1.5%) a dislocation occurred. These patients were compared with a control group matched in number. RESULTS: No difference was found in both groups with respect to age, height, weight, body mass index, and diagnosis. A tendency was found that more males were involved in the dislocation group. There were more previous operations found in this group (p = 0.005). Also significant was the fact, that patients in the dislocation group more often had epidural anaesthesia (p = 0.02), more often the implantation of the smaller 28-mm head (vs. 32 mm) (p = 0.02), and a higher inclination angle of the acetabular component (p = 0.02). No difference was found in terms of the surgeons experience, the postoperative leg length, femoral offset, medialisation, cranialisation and antetorsion of the acetabular component. CONCLUSION: Dislocation after total hip arthroplasty is a multifactorial complication. With the use of the Alloclassic/Zweymuller total hip endoprothesis, a higher risk for dislocation was found for patients with previous hip surgery, the use of the smaller endoprosthesis head, a high acetabular inclination angle, and in the use of epidural anaesthesia. PMID- 11486621 TI - [Long-term results with Zweymuller cement-free Alloclassic stem]. AB - AIM OF STUDY: The aim of our study is to show the long-term results using the Zweymuller cementless Alloclassic stem. METHOD: In our retrospective study 133 from 206 cases (64.4%) were examined after 10.6 years. The examination was done clinically (Scheme after Merie d'Aubigne) and radiologically. The average age was 72 years (48-92 years) at the time of examination. RESULTS: Intraoperative complications occurred in 3 cases (1.5%), 2 were temporary sciatic-nerve paresis and 1 was a stem fissure. Nine cases had been revised (6.7%). In four of these cases the inlay and the head had to be changed, in two cases mechanical loosening of the cup was found; furthermore, there were one removal of heterotopical ossification and one early infection. Stem problems appeared in only one case with deep infection and had been revised twice. There was neither stem sinking nor mechanical loosening in our series. The average polyethylene wear using a ceramic head with 32 mm diameter was 1 mm after 10.6 years. Clinically excellent results were found in 90% of the cases. CONCLUSION: The long-term results using the cementless Zweymuller stem with a rectangular transversal section and easy operation technique over 20 years are excellent Because of the cortical anchorage and the biocompatible titanium alloy the Zweymuller stem can be used in all femur forms and in different age groups. PMID- 11486622 TI - [Long-term results with cement-free total hip prostheses (Zweymuller)]. AB - AIM OF THE STUDY: The purpose of the study is the evaluation of subjective, clinical and radiological long-term results of cementless total hip replacements with "Zweymuller" stems and various cups. METHOD: 113 of 165 consecutively implanted "Zweymuller" stems have been examined clinically and radiologically as well as assessed for subjective satisfaction by questionnaire, 26 additional patients were analysed by questionnaire only. Thus, the follow-up rate is 85.5% with a mean follow-up period of 8.9 years (range 5-12.2 years) as far as evaluation of subjective information including data on failures is concerned. The follow-up results were rated by the scores of Merle d'Aubigne and Harris and radiolucent lines on the X-rays were analysed. RESULTS: 96% of the analyzed patients stated subjective satisfaction. 101 hips caused no or only little pain. The average Harris Hip Score rose from 47.1 preoperative to 86.8 postoperative, the functional value of Merle d'Aubigne rose from 4.9 to 10.1 (maximum 12 points). The survival rate of the stems after 10 years is 96%, the complication rate is very low. 81% of the stems showed a proximal radiolucent line. 3 different kinds of cups were used, 79% of the polyethylene cups and 8% of the titanium cups, respectively, were loosened or had been explanted at the time of examination. CONCLUSIONS: During an observation period of 12 years, the implantation of the "Zweymuller" stem has very successful results. The proximal radiolucent lines should nevertheless cause concerns about the predominantly distal anchoring of the stem. The long-term survival of the stems is much better than that of the titanium cups. PMID- 11486623 TI - [10 years results with a Monobloc hip endoprosthesis cup with multilayer titanium mesh coating for cement-free implantation]. AB - PURPOSE: Can a cementless hemispheric acetabular component which is made of ultra high-density polyethylene (UHMW-PE) and a heat-bounded porous titanium mesh coating be recommended 10 years after implantation? METHODS: Between 1986 and 1988, 279 total hip arthroplasties in which the cementless socket had been used were performed in 261 patients. The results of 145 patients with 159 hip sockets (55.5%) were reviewed clinically and radiologically for comparison after a minimum of 10 years. The clinical and radiological re-examination was documented in a standardized questionnaire in which several scores were integrated. All X rays were stored and analysed by a special hardware and software computer system. RESULTS: The mean age of the female patients (n = 85) was 63.1 years and of the male patients (n = 60) 60.9 years. The mean follow-up period was 10 years and 3 months. The Merle d'Aubigne hip score increased from 10.9 to 16.5 points at the follow-up evaluation. Radiolucent lines and cysts according to the zones of DeLee/Charnley were seen only in 2% of all cases. The mean polyethylene wear (head disarrangement) was measured with 1.79 mm after 10 years and 1.92 mm after 11 years. In total, 16 revisions (5.7%), 13 (4.6%) aseptic and 3 (1.1%) septic, out of 279 implanted sokets had to be performed. In particular, the acetabular cup with a diameter of 48 mm demonstrated a high loosening rate (4 of 9). The 10 year survival analysis (Kaplan-Meler) was measured with 92.8%. CONCLUSION: Our results with a cementless socket with titanium mesh coating show insignificant radiolucent lines and an exceptional high 10-year survivorship in which the particular implant philosophy is of great importance for the long-term success. In conclusion, the acetabular component can be recommended for further implantation. PMID- 11486624 TI - [Survival rate of a cement-free cup of titanium with laser structured surface]. AB - PURPOSE: So far no clinical results concerning the survival rates of cementless pure titanium cups with laser-structured surfaces exist. This study was performed to evaluate our results for the Alphasegment acetabular component (Alphanorm). METHOD: Retrospective analysis of 50 cups in 45 patients was performed (operations performed between November 1995 and January 1998). The average age at the time of the implantation was 65.1 years. Dysplastic hips and revisions were excluded. The survival rate was calculated according to the Kaplan-Meier analysis with the end point defined as the revision surgery of the cup. RESULTS: The average follow-up period was 2.9 (0.6-4.0) years. Two patients (i.e., two implants) died during the observation period, none were lost for follow-up. Six of the cups had to be revised due to aseptic loosening. The survival rate at 3.2 years after implantation was 87%. None of the stems had to be revised. CONCLUSIONS: Further use of the cementless hemispherical pure titanium cup with laser-structured surface cannot be recommended. The observed prevalence of loosening for the Alphasegment acetabular components was higher than for other cups. Close follow-up of patients with this type of implant is necessary. PMID- 11486625 TI - [Thromboembolic complications related to the use of bone cement in hip arthroplasty--pathogenesis and prophylaxis]. AB - INTRODUCTION: Cemented total hip arthroplasty (THA) has to be considered a high risk procedure with respect to cardiovascular complications. The insertion of cement and prosthesis may lead to intravasation of fat and bone marrow into the circulation. METHODS: This article represents a review on the relevant literature about thromboembolic complications associated with the use of bone cement in THA. RESULTS/DISCUSSION: The method of canal preparation and lavage is of paramount importance to reduce the potential embolic load. Additionally, thorough cleaning of the intramedullary cavity (using jet lavage) improves the cement penetration into the bone and increases the shear strength of the bone-cement interface. Modern cementing techniques include the use of high pulsatile lavage, a cement restrictor and cement pressurization. With the application of these techniques a sufficient drainage of the medullary canal should be guaranteed to minimise the risk for thromboembolic complications. During the insertion of cementless implants the intravasation of embolic material seems to be less, but it is likewise possible to have a thromboembolic complication during cementless THA. However, it is not justified as a consequence to use the thromboembolic risk as the main indication for implant choice, in particular as the overall risk of a serious fatal complication is low. PMID- 11486626 TI - [Cement-free diaphyseal fixation principle for hip shaft exchange in large bone defects--analysis of 12 years experience with the Wagner revision shaft]. AB - PURPOSE: Considering the increasing incidence of revision for failed total hip arthroplasty, we evaluated if the principle of uncemented diaphyseal fixation which is realised with the Wagner SL revision stem, can solve the technical problems of significant bone loss of the proximal femur and can offer good preconditions for bony restoration in the medium term. METHODS: A consecutive series of 129 femoral revisions with the Wagner SL revisions stem was evaluated. In 87 cases, the acetabular component was revised simultaneously. The revision was indicated in 97 cases of aseptic loosening, in 13 periprosthetic fractures (12 aseptic and 1 septic hip), in 16 cases of septic loosening and in 3 Girdlestone-hips after chronic deep infection. At the time of revision, the mean age of the patients was 64.9 (36.7-86.3) years. For classification of the preoperative defects, both the classification system proposed by Pak et al. and our own classification system were used. RESULTS: The mean period of time between operation and latest follow-up examination, rerevision, or death of the patient was 5.4 years (0.13-11.7 years). Six revision stems required rerevision because of malpositioning (1), significant subsidence (1), periprosthetic fracture (1), and deep infection (3). The mean Merle d'Aubigne score improved from preoperatively 7.7 points to 14.8 at follow-up. Cumulative survival (end point removal of the stem for any cause) was 94.4 percent at 11.7 years. A clear, good, or excellent bony restoration of the proximal femur was seen in 87.6 percent of the cases. The cumulative survival of the revised acetabular components was 97.7 percent at 11.7 years and for the non-revised cups it was 86.9 percent at 10.9 years. CONCLUSION: Due to the encouraging results with the Wagner revision stem, the principle of uncemented diaphyseal fixation seems to us to be able to solve most of the technical problems in cases of significant bone loss and obviously offers good preconditions for bony restoration. Our own classification system was effective and useful for preoperative planning. PMID- 11486627 TI - [Urinary concentration of collagen metabolites in endoprosthesis loosening]. AB - Aseptic loosening is the most common long-term complication in arthroplasty. Loosening is in every case associated with bone resorption at the interface that leads to bone defects and complicates the revision. The diagnosis of aseptic loosening is based on clinical and radiological evaluation. Especially in clinically asymptomatic cases an early diagnosis with these methods is difficult. In our study we wanted to evaluate the diagnostic value of biochemical markers of the bone resorption in aseptic loosening. We compared 58 patients with proven implant loosening during surgery with 67 patients without clinical or radiological signs of loosening. We measured the crosslinks pyridinoline and hydroxypyridinoline in urine samples. In contrast to Schneider et al. [increased urinary crosslink levels in aseptic loosening of total hip arthroplasty, J. Arthroplasty 1995; 13 (6): 687-692] we found no significant differences between loose and asymptomatic hip or knee prosthesis. Also no correlation between the size of the acetabular defects of loose hip implants and the urinary crosslink excretion was measurable. Our results show no or only little diagnostic value of the urinary crosslinks pyridinoline and deoxypyridinoline in aseptic loosening of total hip and knee arthroplasty. PMID- 11486628 TI - [Epiphyseolysis capitis femoris (ECF) in monozygotic twins]. AB - AIM: The frequency of bilateral slipped capital femoral epiphysis (SCFE) is 5 80%. A set of boy twins developed left-sided SCFE within 2 years. We tried to find a relationship between SCFE and HLA typing. METHOD: We obtained the HLA typing and compared it to previously reported cases. RESULTS: By comparing 14 gene loci we made sure that they are identical twins. In our patients as well as in may of previously reported cases of sets of boy twins HLA phenotype A2 were found, while in the sets of girl twins HLA phenotype A11 and B12 were found. CONCLUSIONS: HLA phenotyping is needed in a larger number of SCFE twin cases to evaluate phenotypic patterns for coincidence to provide a basis for the genetic expression of this condition. PMID- 11486629 TI - [Spondylothoracic dysplasia--a case report of 2 affected sisters with 20 year follow-up]. AB - This is a case report and review of the literature in spondylothoracic dysplasia. Spondylothoracic dysplasia is a genetically transmitted, rare entity characterized by multiple vertebral abnormalities and rib anomalies. Two siblings from one family of normal parents who have four other normal children were affected. There is no history of congenital anomalies in the family. Due to progressing deformity of the spine, a convex epiphyseodesis had to be performed in the oldest girl. The second girl, who has a healthy twin sister, was borne with an inperforated anus and a recto-vaginal fistula. An operation was performed at an early stage. Today, the mature patients are painfree and fully integrated in social life. Spondylothoracic dysplasia is discussed with regard to the hereditary nature, course, and prognosis on the basis of the pertinent literature. PMID- 11486630 TI - [Epidemiology and chrono-epidemiology of distal radius fractures]. AB - The frequency of distal radius fractures of patients treated in the outpatient clinic of the Herz-Jesu-Krankenhaus between Jan. 1997 and Dec. 1999 were evaluated retrospectively. Records of 366 patients (m: 132, f: 234) were assessed. The frequency of distal radius fractures was fitted to a cosine model using non-linear regression in order to detect a circadian or circannual rhythm. The goodness of fit was assessed by means of the F statistics and the coefficient of determination r2. Moreover, spectral analysis was performed. In 1997 135, in 1998 119, and in 1999 112 distal radius fractures were treated. The mean age of the patients was 40.3 ys. (women: 48.8; men: 25.3). The extended wrist was the most common position during Trauma, Accidents at home (41%), during spare time activities (23.8%), and during physical activities (21.6%) were more frequent than accidents during working hours (12.5%) or traffic accidents (1.1%). The latency for definite surgical treatment in our clinic was 8.5 hours (minimum: 15 min; maximum: 7 days and 14 hours). A significant cosine model could be fitted for a period of 24 hours (r2 = 0.82) with a maximum at 15.00 hours. Spectral analysis revealed an ultradian rhythm of approximately 12 hours in addition to the circadian periodicity. No significant circannual model could be fitted (r2 = 0.2), although a slight increase of frequency was found during May and June. In conclusion, the frequency of distal radius fractures shows a clear circadian but no circannual periodicity. PMID- 11486631 TI - [Fracture behavior of AO 3.5 mm cortical titanium screws Synthes screws) combined with LC-DCP plates]. AB - AIM: It was investigated if AO 3.5-mm titanium cortical screws used in combination with LC-DCP plates for osteosynthesis are likely to break during surgery. METHOD: The moments of torque that lead to breakage of the screw were determined experimentally in a hardwood model. The insertion angle was variable: 90 degrees in centric (n = 30) and excentric (n = 30) positioning, 70 degrees (n = 30) and 50 degrees (n = 30). Minimal moments that led to screw breakage were compared to insertion moments that were measured intraoperatively in 7 fracture operations (radial, ulnar, tibial, fibular; 32 screws). RESULTS: Minimal moments that led to screw breakage were 2.6 Nm (90 degrees centric), 2.8 Nm (90 degrees eccentric), 2.7 Nm (70 degrees) and 2.4 Nm (50 degrees). The maximal intraoperatively measured insertion moment was 2.25 Nm (radius). CONCLUSIONS: In this investigation, minimal moments that led to screw breakage in an experimental setting were higher than maximal insertion moments that were recorded during surgery. It is concluded that screw breakage related to the given implant combination is unlikely if correct surgical technique is performed. PMID- 11486632 TI - [Orthopedic aspects of osseous echinococcosis--radiologic diagnosis, current surgery and drug therapy aspects]. AB - PURPOSE: Skeletal manifestation of alveolar (E. multilocularis) or cystic (E. granulosus) echinococcosis is seldom, with an incidence of less than 2% of all cases. By reporting on a rare case of bony manifestation of E. granulosus, we intend to emphasize current standards in surgical and drug management of the disease. Typical radiographic and blood-chemistry findings are highlighted on a 62-year-old Arabiqu female suffering from a cystic echinococcosis with skeletal manifestation in the left ileum and lower lumbar spine. RESULTS: Radiographically typical are fine zones of osteolysis which, at a later stage, show confluence. Due to the rigid structure of cortical bone, cysts tend to grow slowly, and seldom exceed 2 cm in diameter. Only after having broken through the cortical bone, may a massive increase in size be observed. By means of serological tests, it is postoperatively possible to evaluate how successful radical surgery with removal of larvae was. Furthermore, serological tests can be used as a screening method, offering a diagnostic tool that can also utilized in detecting a recurrence. CONCLUSION: The surgical aim in the treatment of osseous echinococcosis should be in accordance to curative tumor surgery. Even after radical removal of the parasites, the WHO suggests an adjuvant chemotherapy with mebendazole or albendazole for at least two years after surgery. In cases where only a palliative treatment is possible, the antihelminthic drug administration can be continuous. PMID- 11486633 TI - [PDS (postdiscectomy syndrome and SPD (postdiscotomy syndrome) in orthopedics]. PMID- 11486634 TI - [Can shoes prevent diabetic foot damage?]. PMID- 11486635 TI - [Experimental study of the effect of acetylsalicylic acid combined with caffeine regarding possible abuse potential]. AB - Caffeine is suspected to initiate the abuse of caffeine-containing analgesics. To clarify this matter the study investigated the effects of aspirin alone and in combination with caffeine on mood and cardiovascular parameters before, during, and after pain induction through mechanical stimulation. 96 healthy middle-aged women--all habitual coffee drinkers--were randomly designed to receive 0 or 500 mg aspirin together with 0, 50, or 100 mg caffeine administered in a double-blind design. Aspirin did not change ratings of mood scales. Caffeine decreased scores on negative mood scales, especially on the anxiety/sadness scale. The combination of aspirin and caffeine failed to increase any score on scales measuring positive moods. Only a few interactions are noticed. The results do not support the suggestion that caffeine increases the potential for abuse of aspirin. PMID- 11486636 TI - [Psychological satiation: a new experimental study of an old construct]. AB - The present experiment on the phenomenon of psychic satiation expands on the work of Karsten (1928) and Lewin (1928). Psychic satiation denotes a loss of intrinsic motivation when the same action is performed repeatedly. Although many studies have shown the high theoretical and practical relevance of this phenomenon, the conditions which lead to or reinforce psychic satiation have rarely been investigated empirically. Based on the concept of psychic satiation as formulated by Karsten and Lewin and refined by Schulz-Hardt, Rott, Meinken, and Frey (in press), we predicted that psychic satiation will increase if the task does not lend itself to being carried out "peripherally" (i.e., as an almost unconscious incidental action) and if it has high personal relevance. These predictions were investigated in an experiment with 66 high-school students who performed different versions of the "Konzentrations-Leistungs-Test" (concentration performance-test, KLT). The results are largely in line with our predictions. In addition, the results indicate that the relation between satiation and performance is moderated by personal relevance and the induced task characteristic. PMID- 11486637 TI - [Response force in probe reaction time tasks: reduction in capacity, response inhibition or bottle neck?]. AB - The probe reaction time (probe-RT) paradigm suggested by Posner and Boies (1971) requires subjects to perform simultaneously a choice reaction task with two fingers of one hand (primary task) and a simple reaction task to randomly presented probes with the other hand (secondary task). The classical interpretation, according to which the RT of the secondary task indexes the capacity demands of the primary task, has been challenged, however. Active inhibition of the response and delay by a central bottleneck have been suggested as alternatives to the capacity account. The present study measured response force in addition to RT to distinguish among the three alternatives. Responses to the probe were clearly more forceful when the probe was presented while the subjects were engaged in the primary task. This result is most compatible with the active inhibition account by Neumann (1987, 1992) combined with an extended motor-readiness model. PMID- 11486638 TI - [Do dissociations between implicit and explicit memory disappear, when reliability of the tests is comparable? An example]. AB - Implicit memory measures tend to be less reliable than explicit memory measures. Consequently dissociations can arise as a consequence of differential reliabilities of the tasks. In the present study the effect of a manipulation of levels of processing in an implicit and an explicit memory task were investigated in a sample of 200 students after establishing comparable reliabilities of the memory tasks. A fragmented object-naming test was used as an indirect test and a recognition task was used as a direct test. The reliabilities of these measures were comparable in all conditions. A main effect of levels of processing, but no interaction between type of test and levels of processing, was found. These findings reinforce the necessity of taking into account the reliability of memory measures for the interpretation of dissociations between explicit and implicit memory. PMID- 11486639 TI - [Success in problem solving as a condition for acquisition of control knowledge in system control]. AB - In an experiment the hypothesis was examined wether successful problem solving facilitates the acquisition of control knowledge in system control. Subjects had to control a complex dynamic system. Success in problem solving was manipulated by imparting structural knowledge. Structural knowledge was assessed by presenting a "pair-task" and by analysing transfer effects in system control. Control knowledge was diagnosed by a recognition task. Subjects taught in structural knowledge learned more about the system and performed better. Effects of instruction on control knowledge did not occur. The results suggest assuming a "look-up table" which stores successful problem solving episodes as well as unsuccessful situations (Berry & Broadbent, 1988). PMID- 11486640 TI - [Association norms for 68 German homonyms]. AB - Norms for 68 German homophonic homographs were collected in a normal population comprising 100 subjects. Frequency and polarity as well as U-scores (index of ambiguity) are presented. 49 of the words revealed a clear dominance for only one meaning. 7 homophonic homographs were balanced, and for 12 words less than 5% of all associations were given for each of the inferior meanings. Studies 2a and 2b are a replication of previous findings on the resolution of semantic ambiguity. Inhibition effects were revealed for discordant triplet conditions which contained a long interstimulus interval between homograph and target, whereas an (insignificant) facilitation effect occurred for trials with a shorter interval. Results of study 2 are in line with context-sensitive models which claim that after the initial activation of all meanings of a homograph a context-guided process leads to a suppression of context-irrelevant homograph information. PMID- 11486641 TI - [Confirmatory information search in simultaneous vs. sequential information presentation]. AB - After having made a preliminary or final decision, people prefer information that supports their chosen alternative to information that conflicts with their choice. Jonas, Schulz-Hardt, Frey, and Thelen (in press) found that sequential presentation of information leads to an even stronger preference for supporting information than the traditional form of simultaneous presentation. Their proposed explanation for this effect was that sequential presentation induces a focus on the prior decision, thereby increasing commitment to this decision. The present experiment was designed to rule out an alternative explanation: Being repeatedly confronted with pieces of information to select from could induce the participants to search for more information than they consider to be necessary, and because less effort is required to process supporting information the additional information requests are predominantly for these supporting pieces of information. To test this alternative explanation, in the present experiment--as in the Jonas et al. (in press) experiments--simultaneous vs. sequential information presentation following a preliminary decision was manipulated. In contrast to the former experiments, this time the number of information requests was fixed: Participants in both conditions had to choose 8 out of 16 pieces of information. The results show that once again a stronger preference for supporting information arises when the information is presented sequentially compared to simultaneously. The alternative explanation mentioned above could thus be ruled out. PMID- 11486642 TI - [Automatic social behavior: how does activation of the concepts of egoism and altruism influence helping behavior?]. AB - An experimental study investigated the influence of priming on helping behavior. A scrambled sentence test was used to prime groups with the concepts of altruism or egoism. In a third group a neutral concept was activated. The results indicated that there is no difference in helping behavior between the altruism and the neutral group. However, in the egoism priming condition participants showed higher latencies and helped less often than participants in both other conditions. PMID- 11486643 TI - The growth effects of gamma-ray irradiation on third-stage larvae of Angiostrongylus cantonensis in snail. AB - In order to clarify growth effects of gamma-ray irradiation on the third-stage larvae of Angiostrongylus cantonensis, snails naturally infected with larvae of A. cantonensis were irradiated with a cobalt-60 source, and the total dosage of exposure was 0.1 kGy. Third-stage larvae were collected from each group of irradiated and non-irradiated snails and then administered to rats. A significant reduction was observed in the number of recovery worms from rats sacrificed at 45 days after infection compared with the control group. Moreover, there was an increase in the percentage of female worms, and the body length of the irradiated group's worms was significantly shorter than that of the control group. In ultrastructural observations by transmission electron microscope, the cuticle was thickened and the subcuticle showed partial vacuolization and separation from muscle in the irradiated group. In addition, the muscles myofibril became shorter, the microvilli of intestinal mucosa were shorter and irregular, the terminal web disappeared, and the phagocytic vesicles, mitochondria, lysosomes and secretion granules decreased in number, respectively. Focusing on the mitochondria of the epithelial cells of the intestine, there were degenerative changes in the irradiated group showing vacuolization. The gonads of female worms recovered from the irradiated group revealed degenerative vacuolization, and the gonads of male worms could not be found. PMID- 11486644 TI - Intrafamilial transmission and risk assessment of HTLV-I among blood donors in southern Taiwan. AB - The human T-lymphotropic virus type I (HTLV-I) is one of the important etiological agents of adult T-cell leukemia/lymphoma and of HTLV-I associated myelopathy or tropical spastic paraparesis. There is still a lack of data concerning HTLV-I transmission by seropositive carriers in Taiwan. We investigated the patterns of HTLV-I intrafamilial transmission in HTLV-I seropositive blood donors and assessed the risk factors of HTLV-I transmission in relatives of HTLV-I carriers in Taiwan. A total of twenty HTLV-I seropositive donors and their 103 relatives were enrolled. Among those 103 relatives, 40 (38.8%) were seropositive for HTLV-I. Their ages ranged from one to 70 years old with a mean age of 31.0 +/- 1.65 year-old. Three of the ten wives of male carriers were HTLV-I seropositive. However, none of the six husbands of female carriers were HTLV-I seropositive. Mother-to-child vertical transmission was found in nine of 48 (18.8%) tested. Significant risk factors of HTLV-I transmission among relatives of HTLV-I carriers were hospital admission, previous transfusion, breast feeding, anti-HCV seropositivity and female relatives of age >/= 30 with odds ratio (OR) of 9.73, 8.64, 4.36, 8.86 and 4.91, respectively (all p < 0.05). Nonsignificant risk factors of HTLV-I transmission were sharing needles, operation history, HBsAg seropositivity and male relatives of age >/= 30. Our findings suggest that mother-to-child and husband-to-wife transmissions are the important forms of intrafamilial transmission of HTLV-I in Taiwan. Screening for HTLV-I in family members of HTLV-I seropositive blood donors may be warranted. PMID- 11486645 TI - Expression of transforming growth factor-beta 1 and alpha-smooth muscle actin of myofibroblast in the pathogenesis of nasal polyps. AB - The pathophysiology of nasal polyps remains unclear, but recent work suggests that many cytokines are produced in nasal polyps (NPs) and that they may play various important roles in the pathogenesis of NPs. Transforming growth factor beta 1 (TGF-beta 1), secreted by many inflammatory cells, is a potent inducer of myofibroblasts. Myofibroblasts express alpha-smooth muscle actin (alpha-SMA) and a source of extracellular matrix (ECM). In this study, we investigated a potential link between inflammation and the growth process in human NPs. Sixteen patients who were affected by NPs and who had undergone functional endoscopic sinus surgery were included in this study. Nasal mucosa of inferior turbinate (NM) of 10 patients who had received rhinoplasty or turbinectomy for other disease was used as the control. alpha-SMA and TGF-beta 1 were detected using immunohistochemistry and the number of labeled cells were counted (alpha-SMA and TGF-beta 1 indices). The expression of alpha-SMA and TGF-beta 1 indices found in NPs and NM was compared using Student's t-test. In our study, alpha-SMA and TGF beta 1 indices were found to be significantly higher in nasal polyps than in nasal mucosa. TGF-beta 1 produced by inflammatory cells can influence the development of myofibroblasts which in turn can induce extracellular matrix accumulation and, therefore, TGF-beta 1 plays a important role in the formation of nasal polyps. PMID- 11486646 TI - Image evaluation of suprazygomatic masticator space lesions. AB - In order to understand lesions involving the suprazygomatic masticator space (SZMS) demonstrated on computered tomography (CT) or magnetic resonance (MR) images, we collected 45 cases of lesions in SZMS. There were hematomas in 26 cases, tumors in 16 cases, and abscesses in 3 cases. The location of these lesions in compartments of the SZMS and adjacent structures was analyzed. Most commonly hematoma was found in the deep loose connective tissue (80.8%), followed by the superficial dense connective tissue (61.5%). Only 2 SZMS hematoma involved the superficial fat pad. Five patients had air accumulation in the SZMS, and all were in the deep fat pad and with maxillary sinus fracture. Of the sixteen cases which were tumors, 12 were extended from adjacent structures, and four were primary tumor. In cases of tumor, compartments of SZMS involved most frequently were temporal muscles (15 of 16 cases), followed by deep fat pad (8 of 16 cases). The connective tissue layer of SZMS is a communication pathway for superficial spread of infection, hematoma, or tumor invasion to or from the scalp and face. Compartments of deep fat pad and temporal muscle are also pathways for the spread of disease to or from the face, intracranium and orbit. The primary tumors in infratemporal fossa and SZMS are rare. Malignant tumors in SZMS usually come from surrounding spaces. PMID- 11486647 TI - Acute leukemia presenting as diabetes insipidus and bilateral exudative retinal detachment--a case report. AB - To report an unusual case of leukemia presenting as both bilateral exudative retinal detachment (ERD) and central diabetes insipidus (DI), we evaluate the clinical hematological records including bone marrow aspirations and CSF tapping, both osmolarity and electrolytes concentration of the serum and urine, brain MRI, fundus photographs and fluorescein angiographs in this 25-year-old female patient. Examinations of peripheral blood and bone marrow aspiration confirmed the diagnosis of acute myelogenous leukemia (AML-M0). Fluorescein angiography (FA) revealed bilateral ERD, dense choroidal leukemia cell infiltration with overlying retinal pigment epithelium (RPE) dysfunction and focal areas of choroidal infarction. Changes in both osmolarity and electrolytes concentration of the serum and urine from vasopressin test supported the diagnosis of central DI. Central DI and ERD may be presenting signs of acute leukemia and both may represent CNS involvement. In our case, dense choroidal leukemic cell infiltration results in devitalization of RPE and choroidal infarction. Leukemic disruption of hypothalamic pituitary area may lead to complete or partial deficiency of antidiuretic hormone (ADH). Rapid improvement in visual acuity and partial symptom relief of DI may ensue from appropriate chemotherapy and nasal DDAVP (1-desamino-8-D-arginine vasopressin) supply. PMID- 11486648 TI - Adrenal cyst--a case report. AB - Adrenal cysts are rare and mostly silent clinically. Herein we report a case of adrenal cyst. A 55-year-old female was incidentally found to have a left suprarenal cystic lesion with a calcified wall by abdominal sonography during a work-up for her epigastralgia and left flank pain. Then, computed tomography (CT) revealed a left adrenal cystic mass with wall calcification, magnetic resonance imaging (MRI) showed left retroperitoneal cystic mass with fluid content, and angiography demonstrated an avascular lesion. Surgical exploration was performed via a flank incision and a calcified cystic adrenal mass was excised. The pathologic diagnosis was adrenal pseudocyst with calcified wall. We discuss the diagnosis and management of adrenal cyst and briefly review the literature. PMID- 11486649 TI - Sequential MR studies of a patient with white matter disease presenting psychotic symptoms: ADEM versus single-episode MS. AB - Acute disseminated encephalomyelitis (ADEM) and multiple sclerosis (MS) are both demyelinating white matter disorders. It is difficult to differentiate ADEM from single episode MS because of the similar clinical presentation, cerebral spinal fluid (CSF) analysis, histological finding and magnetic resonance image (MRI) appearance. We report an ADEM case with unusual clinical presentations of predominant psychiatric symptoms, and relatively long disease course. Initially, we were not able to distinguish it from single episode MS. By means of sequential MR images followed up 2 years and spectroscopy studies, and the dramatic clinical improvement after corticosteroid therapy, ADEM was diagnosed as the disease entity of this patient. In this case report we will present the MR findings of this patient and discuss the differentiation between the ADEM and MS. PMID- 11486650 TI - Percutaneous transluminal angioplasty as an adjunct to treat the carotid cavernous fistula. AB - We report a case of the direct carotid-cavernous fistula (CCF) with high-grade stenosis of the internal carotid artery (ICA) and treatment via the transarterial route after percutaneous transluminal angioplasty (PTA). A 46-year-old man was found to have a CCF after a motorcycle accident. Transarterial embolization was attempted, but it failed due to stenosis of the left cervical ICA. After the procedure of PTA for dilatation of the stenotic ICA, the fistula was successfully obliterated by detachable balloon subsequently. There were no complications or recurrence of the fistula at the three-year clinical follow up. PMID- 11486651 TI - Unrelated donor hematopoietic transplantation. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice for a range of malignant and non-malignant diseases. Unfortunately, fewer than 30% of patients have a human leukocyte antigen (HLA)-matched sibling. Advances in our understanding of the HLA system and the development of large international donor registries are supporting the increasing use of unrelated donors as an alternative source of stem cells. Unrelated donor transplantation, however, is still associated with higher complication rates than in HLA-identical sibling donor transplants. Improvements in graft-vs.-host disease prevention and treatment, new conditioning regimens and better donor selection will likely expand the indications of unrelated donor HSCT in the next decade. PMID- 11486652 TI - Non-myeloablative stem cell transplantation and donor lymphocyte infusion for the treatment of cancer and life-threatening non-malignant disorders. AB - Allogeneic bone marrow or blood stem call transplantation (BMT) represents an important therapeutic tool for the treatment of otherwise incurable malignant and non-malignant diseases. Until recently, autologous and allogeneic bone marrow and mobilized blood stem cell transplantations were used primarily to replace malignant, genetically abnormal or deficient immunohematopoietic compartments, and therefore highly toxic myeloablative regimens were considered to be mandatory for the effective eradication of all undesirable host-derived hematopoietic elements. Our preclinical and ongoing clinical studies have indicated that much more effective eradication of the host immunohematopoietic system cells can be achieved by adoptive allogeneic cell therapy with donor lymphocyte infusion following BMT. Thus, eradication of blood cancer cells, especially in patients with chronic myeloid leukemia and, less frequently, in patients with other hematologic malignancies, can frequently be accomplished despite the complete resistance of such tumor cells to maximally tolerated doses of chemoradiotherapy. Our cumulative experience has suggested that graft-vs.-leukemia (GVL) effects might be a useful tool for the eradication of otherwise resistant tumor cells of host origin. Based on the cumulative clinical experience and experimental data in animal models of human diseases, it appears that the induction of host-vs.-graft tolerance as an initial step may allow the durable engraftment of donor immunocompetent lymphocytes, which may be used for the induction of effective biologic warfare against host-type immunohematopoietic cells that need to be replaced, including malignant, genetically abnormal or self-reactive cells. Based on the aforementioned rationale, we speculated that the therapeutic benefit of BMT may be improved by using safer conditioning as part of the transplant procedure, with the goal being to induce host-vs.-graft tolerance to enable subsequent induction of GVL, possibly graft-vs.-tumor or even graft-vs. autoimmunity effects, rather than attempting to eliminate host cells with hazardous myeloablative chemoradiotherapy. This hypothesis suggested that effective BMT procedures could be accomplished without lethal conditioning of the host, using new well-tolerated non-myeloablative regimens, thus possibly minimizing immediate and late side-effects related to the myeloablative procedures until recently considered to be mandatory for the conditioning of BMT recipients. Recent clinical data presented in this review suggest that effective BMT procedures may be accomplished with well-tolerated non-myeloablative stem cell transplantation (NST) regimens, with no major toxicity. Thus, new NST approaches may offer the feasibility of safer BMT procedures for a large spectrum of clinical indications in children and elderly individuals, without lower or upper age limits, while minimizing procedure-related toxicity and mortality. Taken together, our data suggest that high-dose chemotherapy and radiation therapy may be successfully replaced by a more effective biologic tool, alloreactive donor lymphocytes, thus setting the stage for innovative therapeutic procedures for safer and more effective treatment of patients in need of BMT. PMID- 11486653 TI - Allogeneic transplantation across the HLA barriers. AB - In high-risk acute leukemia patients, a 10-fold increase in the dose of extensively T-cell-depleted hematopoietic stem cells ensures sustained full-donor engraftment of one-haplotype-mismatched transplants without graft-vs.-host disease. Since our first successful pilot study, which exploited the principle of a megadose stem cell transplant, our efforts have concentrated on developing new conditioning regimens, optimizing graft processing and improving the post transplant immunologic recovery. The results so far achieved in more than 100 high-risk acute leukemia patients show that haploidentical transplantation is now a clinical reality. Because virtually all patients in need of a hematopoietic stem cell transplant have a full-haplotype-mismatched family donor, a T-cell depleted mismatched transplant can be offered with curative intent, thus extending allogeneic transplantation procedures to virtually all candidates. PMID- 11486654 TI - Developments in hematopoietic stem cell transplantation. PMID- 11486655 TI - Allogeneic peripheral blood stem cell transplantation. AB - Granulocyte colony stimulating factor (G-CSF)-mobilized peripheral blood stem cells (PBSC) are now widely used instead of bone marrow for autologous transplantation due to earlier hematopoietic recovery after transplant. The low toxicity of G-CSF has prompted phase I and II studies to evaluate PBSC for allogeneic transplantation; these studies have demonstrated that engraftment of neutrophils, red blood cells and platelets is faster with peripheral blood cells compared to marrow. In randomized studies comparing mobilized PBSC and marrow for allogeneic transplantation, most trials have confirmed significantly earlier engraftment with PBSC and similar risks of acute graft-vs.-host disease (GVHD). In some trials, an increase of 10-15% in grade II-IV GVHD has been noted with PBSC. All studies showed a trend towards more chronic GVHD with PBSC. Some randomized studies have shown improved survival and disease-free survival with the use of PBSC due to lowered transplant-related mortality and fewer relapses in recipients of PBSC as a result of improved immune reconstitution and a graft-vs. leukemia (GVL) effect. This survival benefit is most apparent in patients with more advanced hematologic malignancies, but further studies are needed to define the relative benefits of PBSC for patients with less advanced disease. The GVL effect of PBSC is currently being exploited with the use of non-ablative allografts. PMID- 11486656 TI - Results of unrelated umbilical cord blood hematopoietic stem cell transplantation. AB - The number of umbilical cord blood transplants is increasing worldwide. The purpose of Eurocord is to evaluate the results and to compare the outcome of umbilical cord blood transplants with allogeneic bone marrow transplants. Data have been reported to Eurocord by multiple transplant centers. Close links have been established with the cord blood banks through Netcord. Bone marrow transplant data have been provided by transplant centers and also through the European Group for Blood and Marrow Transplantation (EBMT) and International Bone Marrow Transplant Registries (IBMTR). Eurocord has analyzed the outcome of unrelated umbilical cord blood transplants from 121 transplant centers and 29 countries. The results showed that survival with unrelated mismatched umbilical cord blood transplants was comparable to that with unrelated bone marrow transplants. Engraftment with cord blood was delayed, resulting in an increased incidence of early transplant complications. The incidence of acute and chronic graft-vs.-host disease was reduced with cord blood grafts even in human leukocyte antigen (HLA)-mismatched transplants and in adults. In patients with leukemia, the rate of relapse was similar to the rate of relapse after bone marrow transplant. The overall event-free survival with umbilical cord blood transplantation was not statistically different when compared to bone marrow transplants. This large registry study confirms the potential benefit of using umbilical cord blood hematopoietic stem cells for allogeneic transplants. PMID- 11486657 TI - Evolution of women's oral health. AB - The current focus on women's oral health emerged from emphasis on the women's health movement. Provision of high quality oral health care throughout a woman's life span requires the enhancement of multiple factors. Future progress will depend on partnering among various constituencies; education at the predoctoral, postdoctoral, and continuing education levels; expansion of the scope of gender specific research; and development of innovative modalities for the diagnosis, prevention, treatment, and utilization of clinical dental services for women. PMID- 11486658 TI - Genetic influences in women's oral health. AB - Variations in the genetic code have been described in an ever increasing array of conditions. This article focuses on a select group of disorders chosen for their relevance both to women and the dental practitioners who care for them. Many of these disorders also illustrate important advances in understanding the complex interaction between an individual's genetic code make-up and the surrounding environment. PMID- 11486659 TI - Maternal oral health. AB - Maternity changes a woman's body and can have an effect on fetal oral health. Prenatal counseling can improve oral health of both mother and child. Maternal diet, self-care, and lifestyle can lead to permanent changes in the offspring's oral health. Transmission of maternal oral microflora and use of fluorides are important in a baby's oral health early in life. PMID- 11486660 TI - Psychosocial issues in women's oral health. AB - Oral health is a significant issue for women because of its association with overall health and quality of life. There is growing evidence that the casual factors and consequences of oral disease are not only physical, but are also social and psychologic. As the importance and implications of the psychosocial factors become better understood, a more comprehensive model for treatment is developing. Accepting this role can lead dentists to a more multidisciplinary approach to successful treatment for their female patients. PMID- 11486661 TI - Eating disorders in women's oral health. AB - Eating disorders such as anorexia nervosa, bulimia nervosa, and binge-eating disorder are a serious concern in women's oral health and a clinical challenge to dental professionals. Each of these eating disorders presents with unique patterns of psychologic, medical, and dental characteristics. Appropriate dental treatment is based in the multidisciplinary facets of these conditions. The dental team should be mindful that individuals who suffer from these disorders may relapse into previous negative eating behaviors. The knowledgeable dental professional may be able to intercept these habits through regular recall intervals and thorough examination. PMID- 11486662 TI - Women's oral health across the lifespan. AB - To address women's oral health conditions, it is important that oral health care providers understand basic female physiology and reproductive biology and consider the complex interaction between the environment, biology, and psychosocial development of women. Most oral health issues are not gender specific but rather are aspects of conditions that may affect women differently. Oral health care providers should recognize that using this information to provide competent care requires knowledge, attitudes, and behaviors that are culturally and gender-sensitive. Recognizing the potential for gender-based differences in oral health and disease processes is a first step to becoming gender-competent. PMID- 11486663 TI - Oral health issues for women athletes. AB - More women are participating in sports at all levels. This article presents information for dental professionals to enhance awareness of emerging issues in women's oral health, with specific emphasis on female athletes. These issues include the prevalence, prediction, and prevention of sports-related traumatic orofacial injuries as well as fads and habits such as tongue piercing, smokeless tobacco, eating disorders, and performance-enhancing drugs. PMID- 11486664 TI - Gender differences in special needs populations. AB - The August 2000 Surgeon General's report confirmed that neglected, vulnerable populations in the United States are not adequately receiving oral health services. The silent epidemic of dental and oral diseases in population groups such as persons with MRDD and the geriatric population results in a diminished quality of life. This article presents a study and research design intended to gather data and report the results as related to the needs of a population with MRDD. The Surgeon General stated that oral health issues in relation to gender have not been explored adequately. The data that were collected add information on health, [table: see text] disease, and health practices in a neglected population, helping in part to address this important concern. The analysis of study data statistically confirmed certain differences between women and men in this special needs population. Women and men experienced a similar number of filled surfaces and missing teeth. Reports of oral pain and daily tooth cleaning were also similar. The rates of fluorosis and the need for urgent treatment differed by gender, as did the number of persons missing a combination of anterior and posterior teeth. In this population men exhibited a greater frequency of past oral injury than women and a greater burden of untreated caries. Men also exhibited a greater frequency of gingival signs, indicating a possible lack of attention to proper oral hygiene; this finding was coupled with their greater frequency of need for urgent dental treatment. Although some reasons for these findings can be postulated, more research into the causes is warranted. Those findings can then form the basis for sound policy decisions that will improve the oral health of both men and women with special needs. Barriers to improved oral health, whether they result from problems with access to providers, from attitudinal problems inherent in the providers themselves, the patient, or the patient's caretakers, or from a possibly gender-biased database, must be effectively addressed. Such barriers can be removed through increased knowledge that takes into account any physiologic and psychosocial differences in men and women, particularly in the long-neglected segments of the population--the elderly and the developmentally disabled. PMID- 11486665 TI - Esthetic facial surgery for women. AB - Women of all ages are increasingly seeking facial cosmetic surgery for personal and professional reasons. A wide array of surgical procedures is available to address each woman's specific request. Esthetic facial surgery and cosmetic dentistry may complement each other to provide dramatic results. PMID- 11486666 TI - Maintaining women's oral health. AB - Women must adopt health-promoting strategies for both general health and the oral cavity, because the health of a woman's body and oral cavity are bidirectional. For general health-maintenance strategies, dental practitioners should actively advise women to minimize alcohol use, abstain from or cease smoking, stay physically active, and choose the right foods to nourish both the body and mind. For oral health-maintenance strategies, dental practitioners should advise women on how to prevent or control oral infections, particularly dental caries and periodontal diseases. Specifically, women need to know how to remove plaque from the teeth mechanically, use appropriate chemotherapeutic agents and dentifrices, use oral irrigation, and control halitosis. Dental practitioners also need to stress the importance of regular maintenance visits for disease prevention. Adolescent women are more prone to gingivitis and aphthous ulcers when they begin their menstrual cycles and need advice about cessation of tobacco use, mouth protection during athletic activities, cleaning orthodontic appliances, developing good dietary habits, and avoiding eating disorders. Women in early to middle adulthood may be pregnant or using oral contraceptives with concomitant changes in oral tissues. Dental practitioners need to advise them how to take care of the oral cavity during these changes and how to promote the health of their infants, including good nutrition. Older women experience the onset of menopause and increased vulnerability to osteoporosis. They may also experience xerostomia and burning mouth syndrome. Dental practitioners need to help women alleviate these symptoms and encourage them to continue good infection control and diet practices. PMID- 11486667 TI - Women's health and oral health implications of the curriculum study. AB - This article discusses the effect of medical school and dental school curriculum surveys, which allowed interdisciplinary analysis of the status of women's issues in the health profession. With this documentation of the status of women's health and oral issues, changes in the curriculum can now occur to close the gaps in education and training exposed in the surveys. Changes in the curriculum are aimed at improving clinical practice by practitioners and lowering barriers to care experienced by women. These changes must be incorporated into not only the medical school and dental school curriculums, but also into the practices of the current health care practitioners to be effective. PMID- 11486668 TI - The patient care development programme: organisational development through user and staff involvement. AB - A number of approaches have been developed in recent years to try effectively to engage service users in the process of planning and delivering health-care services. The consumerist methodology for the strategy described in this paper was designed to maximise staff involvement in capturing user views, in order to develop services at a district general hospital. This strategy--the Patient Care Development Programme (PCDP)--provides a framework for both staff and patient involvement in shaping and influencing the development of health-care services. Uses the findings from applying the strategy to modify care packages, roles, skills, layouts, protocols and procedures, in response to both the "shortfalls" and the service strengths that the patient's view uncovers. Discusses the results of an evaluation of the programme which has been replicated in another part of the UK. The PCDP now forms part of a clinical governance framework and is being used to develop multi-agency integrated care pathways. PMID- 11486669 TI - Causes of cost inefficiency in HMOs. AB - Estimates a total effects cost function using a national 1994 health maintenance organization (HMO) data set to examine and update findings related to HMO efficiency. The cost function controls for ownership characteristics (profit status and ownership), size, enrollment diversity, regional location, product diversity, model type, payment characteristics, and years of operation. While not explicitly controlling for quality or acuity, measures of plan and enrollee diversity help control for aculty and quality. Results show that most of the difference in cost efficiency between HMOs is explained by factors specific to the HMO, including efficiencies of scale and scope, lower levels of Medicare patients, and efficient levels of capital. The study also shows that for-profits are more efficient than non-profits because they rely less on withhold pools to control costs. Limitations of the study include weak controls for quality of care, and limited data related to payment characteristics. PMID- 11486670 TI - Clinical governance is "ACE"--using the EFQM excellence model to support baseline assessment. AB - The introduction of clinical governance in the "new NHS" means that National Health Service (NHS) organisations are now accountable for the quality of the services they provide to their local communities. As part of the implementation of clinical governance in the NHS, Trusts and health authorities had to complete a baseline assessment of their capability and capacity by September 1999. Describes one Trust's approach to developing and implementing its baseline assessment tool, based upon its existing use of the European Foundation for Quality Management (EFQM) Excellence Model. An initial review of the process suggests that the model provides an adaptable framework for the development of a comprehensive and practical assessment tool and that self-assessment ensures ownership of action plans at service level. PMID- 11486671 TI - Patient participation requires a change of attitude in health care. AB - A partnership is founded on equality and mutual respect. In health care there is no balance of power between the patient and the health-care professional. To be able to create healthy partnerships we can aim for a balance by focusing on the interpersonal relationship between partners. To understand the system and be able to change it we have to see it from the patient's point of view. This implies a change of attitude in health care. Each of us have to select, plan and execute our own behavioural changes. We also have to create a system based on the premises of the consumers. Establishing a patient/relative panel is an example of creating an arena for building partnerships with patients. By combining professional knowledge with systematic input from experienced consumers the hospital organization can get access to valuable knowledge and insight to improve the care for the patients. PMID- 11486673 TI - The customer chain. PMID- 11486672 TI - Achieving clinical governance in Women's Services through the use of the EFQM Excellence Model. AB - Following a brief explanation of the concepts inherent within the European Foundation for Quality Management Excellence Model, the experience of using the framework as a mechanism for delivering clinical governance is described. The framework was utilised by a Women's Services Directorate of an acute National Health Service Trust in the UK, who concluded that the Model was an ideal tool for supporting the delivery of clinical governance. However, this was only the case when a number of factors were taken into consideration. For instance, the Directorate found that the change programme required a phased implementation process, sound leadership, expert facilitation, good information systems, numerous training and development opportunities for managers, teamwork and the application of best practice in relation to project improvement teams. Moreover, the absence of all the aforementioned ingredients had the potential to compromise any successful outcome. PMID- 11486674 TI - The economics of quality--a practical approach. AB - How much time and money should we devote to quality activities? Will the results be worth the resources which we invest? How can we calculate the return on our quality investments? These questions are raised by managers, clinicians and policy-makers, but they are rarely answered, which may explain the loss of credibility of some quality activities. This paper gives examples of the cost of poor quality, describes and illustrates a simple method for quality costing, and discusses the economics of quality. It considers why there is little research and teaching about the subject, and proposes how managers and clinicians can take a more economically-informed approach to quality in public healthcare. PMID- 11486675 TI - Re-engineering pharmaceutical care: towards a patient-focused care approach. AB - As healthcare reform takes shape, many challenges face hospital pharmacists. An opportunity exists to combine the principles of patient-focused care and pharmaceutical care to redesign the role of pharmacy. To achieve this objective, pharmacy departments should adopt business concepts such as process re engineering. Process re-engineering is a change management tool which aims to produce dramatic improvement in performance measures by re-designing the process. The goal of restructuring is to increase the amount of time pharmacists spend providing pharmaceutical care to patients. The pharmaceutical care concept is a method of delivering pharmaceutical care services that match individual patient needs with the services provided. This article describes many hospital pharmacy department transitions to a patient focused care environment by adopting the patient focused care concept and the process re-engineering to improve the quality of patient care through systems improvement. PMID- 11486676 TI - Inpatient care quality: analyzing Swedish hospitals with stroke as a tracer. AB - Mortality statistics are an important source of information concerning variations in time and place, identification of risk factors and the evaluation of treatment programs. In this study, a new death certificate was completed "blind" on the basis of hospital records from the last episode of care, across a random sample of 1,376 cases. The results showed that the overlap between the official register's underlying cause of death and that of a panel was 72 per cent at the three-digit level. The official underlying cause of death from cerebrovascular diseases (CVD) was 72 cases in this sample, while 93 were deemed to have CVD by a panel. Additionally, of the 1,233 cases originally reported as non-CVD, the panel deemed non-CVD to be the true underlying cause in 1,176 cases. The paper concludes that CVD was most often correctly reported as the underlying cause of death in the investigated ages up to 75 years but plain differences were found between specialities and in different hospital size. PMID- 11486677 TI - Quality improvement in the healthcare industry: some evidence from Singapore. AB - Presents an exploratory field research on all hospitals in Singapore, highlighting the different routes hospitals have adopted in pursuing their corporate quality journey for the new millennium. In general, both continuous improvement and innovation-based approaches have generated cost and time savings and helped to streamline work processes. However, the initial survey results show that innovation-based programs require a longer time frame for implementation, are more prone to resistance to change and suffer from program failure. Also, large hospitals and public hospitals are more inclined to implement innovation based approaches while medium-sized hospitals tend to use continuous improvement as a medium for quality improvement. PMID- 11486678 TI - Severe anaemia during pregnancy in Kisumu District of Kenya: prevalence and risk factors. AB - Anaemia during pregnancy is an important contributor to maternal mortality and morbidity in Kenya. A prospective study was conducted in Kisumu District, a region characterized by high incidences of maternal and infant mortality, to determine the levels and prevalence of maternal complications. Four health facilities were purposely selected to act as sentinel centres from urban and rural clusters. All the obstetric-related cases were recorded between January and July 1997, and the outcomes of the pregnancies were recorded. Details on the respondents socioeconomic, demographic, biomedical and environment characteristics were also recorded. A total of 1,455 cases were recorded, of which 59 percent experienced obstetric-related complications. Of those with complications, 22 percent were suffering from severe anaemia. Among other things, the study reveals that anaemia prevalence is determined by maternal and environmental factors. These include poor pregnancy care, illness during pregnancy, socioeconomic conditions of the mother and the sanitary conditions of the household. Policy measures aimed at managing anaemia should seek to address all these factors. PMID- 11486679 TI - [Clinical and epidemiological aspects of age-related hearing loss in the Veneto region]. PMID- 11486680 TI - Synovial osteochondromatosis of the hip joint. Etiology, diagnostic investigation and therapy. AB - Synovial osteochondromatosis (Reichel's syndrome) of the hip joint is an unusual entity that is difficult to diagnose in the initial stages. In the beginning the condition is characterized by recurrent pain in the affected joint with negative radiographic and MR findings. The first stage shows only synovial reaction without loose bodies. In the second stage radiography is negative because of cartilaginous bodies, and even in the third stage radiography is negative in one third of cases although osseous bodies are present. After diagnosis surgery is indicated to prevent damage to the joint. We review the etiology, clinical findings and differential diagnosis especially types of secondary synovial chondromatosis and discuss ultrasound, radiologic, MR and scintigraphic features and laboratory findings. We describe the surgical treatment and postoperative treatment of patients with synovial osteochondromatosis with moderate signs of secondary osteoarthritis. PMID- 11486681 TI - Fractures of the distal radius. Current concepts for treatment. AB - The authors review the treatment of fractures of the distal radius, based on their experience and from data in the literature. The choice of a treatment for any given fracture must take into account first of all the stability of the fracture. The best results are achieved in stable fractures. Only minimally displaced distal radius fractures can be treated functionally. However, a plaster cast for one week is indicated for the comfort of the patient. In displaced but stable fractures both closed reduction and percutaneous fixation are indicated. In case of closed reduction, the plaster cast should be applied for 5 to 6 weeks with an above-elbow cast for 3 weeks. Percutaneous fixation gives the best results in extraarticular fractures in younger patients. Because of its simplicity however, it should not be ignored in the elderly osteoporotic patients. In the authors' experience, both techniques were only used for extraarticular fractures. Good and excellent results were found in the closed reduction and plaster cast group in 74% of the patients; the Kapandji technique gave 75% good and excellent results. These results are in line with other findings which show that, for simple fracture types, the Kapandji technique and closed reduction seem to give similar results. External fixation is widely used for intra-articular comminuted fractures. Dynamic external fixation does not show any advantage over static devices. Additional K-wires or bone grafting may be necessary. External fixation gives superior results to plate and screw fixation. Internal fixation should be reserved for fractures with ventral comminution or severe displacement with unacceptable reduction by closed or minimally invasive techniques. PMID- 11486682 TI - Biomechanical evaluation of Hackethal's intramedullary bundle pin fixation of humeral neck fractures. AB - Humeral neck fractures can be stabilized using a bundle of intramedullary pins as described by Hackethal. In order to decrease the risk of pin migration, packing of the medullary cavity with as many pins as possible is sometimes recommended, but others believe that stability can be decreased by destruction of cancellous bone in the humeral head by a large bundle of pins. A surgical neck fracture was created with a saw in 30 frozen cadaveric humeri. Bone quality was evaluated by radiography and densitometry. Fractures were stabilized using Hackethal's technique of retrograde intramedullary pinning with varying numbers of 2.5-mm diameter pins; increasing torsion or bending moments of force were then applied to the bones studied. Stability was found to improve with an increasing number of pins and with higher humeral head density. Based upon these findings, the use of a large number of pins is recommended to reduce the risk of pin migration. Up to eight pins, the risk of destruction of cancellous bone in the humeral head appears very low. PMID- 11486683 TI - Surgical treatment of three-part proximal humeral fractures. AB - The majority of proximal humeral fractures may be managed with nonsurgical treatment. However, three-part displaced fractures require surgery with two goals: obtaining an anatomic reduction and providing enough stability to allow an early rehabilitation program. This article describes a simple technique of internal fixation with Kirschner wires that can be applied in fracture dislocations and three-part fractures of the proximal humerus. A retrospective study of 29 consecutive cases of three-part fractures of the proximal humerus treated with this procedure is presented. Excellent or satisfactory results were achieved in 79.3% of cases using Neer's criteria at minimum 12 months follow-up. Eighteen patients (62.1%) had no pain or had only mild pain not interfering with daily activities. Twenty-four shoulders (75.9%) had more than 130 degrees active elevation, and 15 patients (51.7%) had at least 130 degrees active abduction. The complications were one avascular necrosis, one osteitis and one early loss of reduction that required a new operation. In conclusion we recommend this technique for operative treatment of three-part fractures of the humerus because of its simplicity and satisfactory results. PMID- 11486684 TI - Long-term results of the Boytchev procedure for the treatment of recurrent dislocation of the shoulder. AB - The authors present the long-term results obtained with the Boytchev technique in the treatment of recurrent anterior dislocation of the shoulder. Recurrence, clinical outcome and functional limitations were evaluated. A total of 27 shoulders in 25 patients were studied. The average follow-up period was 13.3 years. There were no immediate neurovascular complications and no residual pain. The average recovery time was 4.3 months. Dislocation recurred in five shoulders (18%) and 18.5% of the patients were left with restricted mobility. All patients resumed their previous employment although sporting capacity recovered to pre injury levels in only 9 of 15 patients. The results were classified as good in 18.5% and as excellent in 40.7% of the patients, but the remaining 40.7% had a fair or poor result. In conclusion, the recurrence rate is high and nearly 20% of the patients are left with limited shoulder mobility. Furthermore, although all the patients were able to return to their job, only 60% of those engaged in sports regained their previous level. This technique, although attractive, is better avoided. PMID- 11486685 TI - Modifications to the mechanical behavior of the wrist after fracture of the scaphoid. Modeling by finite element analysis. AB - Although based on their long-term clinical evolution there appeared to be no doubt that fractures of the scaphoid modify the mechanical behavior of the carpus, the mechanisms of these modifications have not yet been investigated. This study based on finite element analysis provides insight into the sequence behind the onset of arthritis of the wrist, highlighting the existence of pressure peaks at the nonunion and at the midcarpal interface (scaphoid-capitate and lunate-capitate). This evidence explains the clinical evolution of nonunions of the scaphoid. Our study indirectly demonstrates the role played by the scaphoid within the wrist as a force transmission column. The use of finite element analysis for the modeling of simple or complex osteoarticular systems may prove to be a highly useful tool for the understanding of these mechanisms. PMID- 11486686 TI - Metacarpal lengthening by distraction osteogenesis in childhood brachydactyly. AB - Metacarpal lengthening was performed by callus distraction in six children with brachydactyly. Four of the patients were girls and two were boys, with ages ranging from 10 to 14 years (mean age: 12 years). Four of the patients had brachydactyly resulting from injury to the hand and damage to the epiphysis related to infection, while the other two had congenital brachymetacarpia. Two patients had short thumbs, two had short index fingers, and the remaining two had short ring fingers. General anesthesia with a pneumatic tourniquet was used during surgery for all patients. Distraction was initiated on the sixth day following osteotomy, and distractions of 0.25 mm were carried out at 12-hour intervals. Adequate length was achieved in an average of 37 days (32-44), and adequate union of the bone in an average of 12.2 (11.5-15) weeks. No bone grafting was done. In one patient with congenital brachymetacarpia, it was not possible to achieve the desired length because of bending in the Kirshner wires, but union of the bone occurred without difficulty, and the outcome was satisfactory. Although single-stage metacarpal lengthening by callus distraction is time-consuming, it is a simple procedure without complications, which can be used in children with brachydactyly. PMID- 11486687 TI - Neck-shaft angle remodelling after derotation varus osteotomy for severe Perthes disease. AB - Sixteen patients with a severe form of Perthes disease who underwent containment derotation varus osteotomy (DVO) of the proximal femur were studied retrospectively for the neck-shaft angle (NSA) remodelling. Analysis of results after a mean follow-up of 7 years (minimum of 5 years and maximum of 8.7 years) showed mean preoperative, immediate postoperative and the latest follow-up neck shaft angles (NSA) as 135 degrees, 110 degrees (none had varus less than 90 degrees) and 125 degrees, respectively. A statistically significant relationship between NSA remodelling and the immediate post-op NSA (P = 0.0035) was established. Patients with smaller postoperative NSA showed better remodelling compared with those who had higher degrees of NSA immediately after the surgery. No significant relationship was found between the degree of NSA remodelling and the patient's age at diagnosis, age at surgery, severity of Perthes or the Stulberg grading at maturity. The authors conclude that up to 90 degrees or more varization after DVO for Perthes disease can remodel with time, and the remodelling process is independent of the patient's age at diagnosis, age at surgery, or severity of Perthes or Stulberg grading at maturity. PMID- 11486688 TI - Repair of chronic ruptures of the anterior cruciate ligament using allograft reconstruction and a ligament augmentation device. AB - A prospective study was performed to determine the effect of a ligament augmentation device (LAD) on the replacement of the anterior cruciate ligament (ACL) using tendon allografts. Twenty-five patients were followed for 66 to 98 months after tendon allograft replacement with LAD reinforcement for ACL rupture. The evaluation was done using the form of the International Knee Documentation Committee (IKDC), the Lysholm score and the Tegner scale. Two patients sustained a rerupture after major injury. Three other multiply-injured patients scored poorly because of associated injuries and fractures (IKDC grade D). Four patients scored normal (grade A), 12 patients nearly normal (grade B), and 5 patients abnormal (grade C). The Lysholm score showed 14 excellent (average 96), 5 good (average 86) and 4 fair results (average 76). Three patients with excellent results were IKDC grade C, solely because the xrays showed slight narrowing of the medial joint line, which might indeed indicate future problems. On the Tegner scale, the sports level decreased by an average of 1.4 points (from 7.25 to 5.83). Only five patients showed an anteroposterior displacement of more than 3 mm, of which only one was in the grade C group. PMID- 11486689 TI - Popliteal artery injuries associated with fractures and dislocations about the knee. AB - The authors present the results of a retrospective review of popliteal artery injuries associated with fractures and dislocations about the knee. They treated 41 patients with popliteal artery injuries associated with either fractures about the knee or knee dislocations. Thirty-five of the patients were males, 6 females; the mean age was 23 years. The delay before accessing the hospital was 17 hours (range: 3 hours to 10 days). Thirty-two fractures were open. Together with the vascular injury, 12 femoral fractures, 20 tibial and fibular fractures, 5 knee dislocations, 4 femoral + tibial fractures were identified. Twenty-three patients underwent external fixation, 8 internal fixation, 6 plaster cast immobilization, 4 minimal osteosynthesis and plaster cast immobilization. The arterial injury was treated by end-to-end anastomosis in 5 cases, saphenous vein anastomosis in 29 cases and thrombectomy in 7 cases. Nine patients were amputated. Delay in surgery, blunt trauma, extensive soft tissue defect and bone fracture or dislocation, are associated with high amputation rate following popliteal artery injury. The influence of each of these factors alone on the amputation rate could not be evaluated in this study, as no statistically significant correlation could be demonstrated. PMID- 11486691 TI - Short amputation stump lengthening with the Ilizarov method: risks versus benefits. AB - The authors point out the indications for lengthening of short amputation stumps. Referring to their own experience they discuss the problems and pitfalls related to this surgical procedure. Sufficient soft tissue coverage is the most important factor for a good outcome whereas limitation of adjacent joint function may have a detrimental effect on the clinical result. PMID- 11486690 TI - Use of gracilis muscle free flap for reconstruction of chronic osteomyelitis of foot and ankle. AB - Despite low donor-site morbidity and a straightforward dissection, the gracilis muscle flap is still for many surgeons a second choice in microsurgical reconstruction of the lower extremity in cases of osteomyelitis. They underscore the difficulty of the procedure, and the problems of insufficient muscle volume and a small sized vascular pedicle. The aim of this study was to assess the reliability of the gracilis muscle free flap in the treatment of osteomyelitis of the foot and ankle. Between 1992 and 1999, 12 consecutive cases (age 9 to 71 years) of osteomyelitis of the foot and ankle were treated using a skin-grafted gracilis free muscle flap. Criteria for osteomyelitis were the presence of exposed bone, positive cultures and bone scans. The wound defect surface ranged from 9 to 90 cm2 (mean 50.5 cm2). Six flaps were applied on the weight bearing area of the foot. Flap harvesting time never exceeded 30 minutes. The mean follow up is 15 months (range 2 to 60 months). All flaps survived completely. Secondary skin grafts were needed in two cases. One hematoma was noted at the flap donor site. Two patients (18%) had persistent osteomyelitis due to insufficient debridement in the presence of what appeared to be extensive bone involvement. Attempt to salvage the extremity was first performed but ultimately led to amputation. No patients complained of any donor site morbidity. Failure to cure the osteomyelitis was never caused by inadequate flap coverage. Gracilis muscle flap reliability in terms of vascular supply and ease of dissection made it our first choice in osteomyelitis of the foot and ankle. In the presence of extensive bone involvement, complex bone reconstruction is necessary to avoid amputation. PMID- 11486692 TI - Injuries from palm tree thorn simulating tumoral or pseudotumoral bone lesions. AB - Three cases of bone changes caused by foreign bodies that appeared to be tumoral lesions or pseudotumors, were observed in young male patients who presented with pain, localized inflammation, and radiographic and bone scan findings suggestive of tumoral or pseudotumoral lesions. Accurate diagnosis was made at surgery when the foreign body was retrieved. Following removal of the foreign body, postoperative recovery was satisfactory. The common causative agent in all these cases was a palm tree thorn. PMID- 11486693 TI - Isolated fracture of the capitate with proximal pole dorsal dislocation. A case report. AB - Isolated fractures of the capitate are uncommon. We report a rare case of isolated fracture of the capitate with dorsal dislocation of the proximal pole. After open reduction and K-wire fixation the fracture united, and a full range of wrist motion was achieved. No signs of avascular necrosis were observed after 3 years. PMID- 11486694 TI - Stress fracture of the femoral neck in a child. Case report and review of the literature. AB - Fatigue fractures generally appear in normal bone following increased repetitive activity or strenuous exercise, when imbalance occurs between applied force and elastic resistance. Femoral neck fatigue fractures are most common in athletes and young military recruits; such fractures in children with open physes are very rare. This paper reports a fatigue fracture of the femoral neck in an 8-year-old girl presenting with pain in the anterior aspect of the right thigh, of one month duration and without evidence of previous injury. Bed rest and relief of weight bearing are indicated for nondisplaced compression fractures. Surgical treatment is indicated for tension fractures. In most cases, the outcome is favorable, and complications are rarely reported. PMID- 11486695 TI - Chondroblastoma of the patella associated with an aneurysmal bone cyst. AB - Chondroblastoma is a rare, benign tumor of bone, accounting for about 1% of all bone tumor cases. It tends to affect the epiphyseal ends of long bones, most often in males during the first and second decades of life. It has well characterized radiographic and histologic features but despite its histologically benign appearance a few cases of metastases have been reported. Local recurrences after curettage and bone grafting occur in 11% to 25% of cases. The features of a patellar chondroblastoma are the same as for other locations. In reviewing the literature we found an unusually high male-to-female ratio. It is interesting that the usual treatment of the patellar chondroblastoma has been patellectomy, whereas curettage and bone grafting has predominated in the other locations. We present a computer tomography and magnetic resonance imaging study of a case of chondroblastoma of the patella associated with an aneurysmal bone cyst. To our knowledge, it is the seventh case reported and the second with computer tomography and magnetic resonance imaging studies. We also review and discuss in detail all the cases of patellar chondroblastoma that we found in the literature. PMID- 11486696 TI - Subungual glomus tumor of the hallux. A case report. AB - The authors report a rare case of subungual glomus tumor in the right hallux, which was excised with complete relief of symptoms. They provide a brief description of histological features and review the literature. PMID- 11486697 TI - Pneumoencephalomeningitis secondary to infected lumbar arthrodesis with a fistula: a case report. AB - Pneumocephalus associated with spinal problems is very rare. Association with encephalomeningitis secondary to a fistula after an infected elective lumbar spine fusion has not been previously reported. The authors report a case in which the clinical onset of pneumoencephalomeningitis occurred after an airplane flight. CT-scan and lumbar puncture were used to make diagnosis; the treatment was based on parenteral antibiotics. The symptoms and signs of infection and neurological deficit resolved but the fistula remained. Diagnosis in such cases must be based upon CT-scan and lumbar puncture. Treatment should consist of systemic antibiotic therapy. Surgical management of infection and fistula is desirable, should the status of the patient allow such a treatment. In any case, as airplane flights in such cases may predispose to pneumocephalus, patients with an infected CSF fistula should avoid airplane flights until the problem is solved. PMID- 11486698 TI - Continuous intra-articular patient-controlled analgesia in a cancer patient with a pathological hip fracture. A case report. AB - Pain management is one of the major goals to achieve in treating terminal patients. Managing severe pain with drug therapy by using a combination of narcotics, non-narcotics, and adjuvant drugs provides the best results for most patients. In patients with fractures due to bone metastases there is a relative indication for operative treatment if the patient is expected to survive the operation and to recover in the postoperative period. In this case report a terminal 48 year-old female with a pathological femoral neck fracture is presented. No operative treatment was offered because of her bad general status. Pain control was maintained by continuous administration of local analgesics directly into the intra-articular femoral space, through a system of patient controlled analgesia (PCA). The visual-analogue pain scale (VAS), which was maximal before beginning of this treatment, was markedly reduced during the treatment, and was again elevated to maximum after the intra-articular catheter was removed. Although continuous regional analgesia to a fractured bone which involves the joint is not a generally acceptable method and carries some risks it has a role in a selected group of patients. PMID- 11486699 TI - Effect of treatment on health-related quality of life in acquired immunodeficiency syndrome (AIDS)-related Kaposi's sarcoma: a randomized trial of pegylated-liposomal doxorubicin versus doxorubicin, bleomycin, and vincristine. AB - The purpose of this study was to determine whether health-related quality of life (HRQL) would be improved in patients with acquired immunodeficiency syndrome (AIDS)-related Kaposi's sarcoma treated by pegylated-liposomal doxorubicin (PLD) as compared to those treated by a conventional combination of doxorubicin, bleomycin, and vincristine (ABV). One hundred thirty-three patients received PLD and 125 patients received ABV every 2 weeks with a planned total of 6 cycles. Patients completed a 30-item AIDS-related HRQL questionnaire before beginning treatment (baseline), every 2 weeks while on treatment, and about 21 days after the end of treatment. Twenty-two items, involving nine domains, were analyzable. While on treatment, PLD-treated patients with partial clinical responses achieved statistically significant greater improvement (compared to baseline) in general health than did ABV-treated patients with partial clinical responses (rho = 0.008). By the end of treatment, the overall group of patients receiving PLD showed statistically significant greater improvement in pain and energy/fatigue than did the group receiving ABV (rho = 0.01-0.002). In addition, duration of clinically significant improvement in global QL was longer in the PLD arm. PMID- 11486700 TI - Long-term effects on the intelligence of children treated for acute lymphoblastic leukemia. AB - The purpose of this study was to investigate whether the intelligence quotient (IQ) in children treated for leukemia decreases in the years following whole brain irradiation. Twenty-seven leukemic children were assessed following a mean time lapse between radiotherapy and IQ measurement of 9 years. The IQ test used was the Hamburg Weschsler Intelligence Test for Adults. The IQ results did not differ significantly, p > 0.05, from the IQs of the general population. It was found that age and dose were not predictors of a decrease in IQ. The only predictor was time lapse between irradiation and IQ measurement, which we found to be indicative of an IQ decrease even after 9 years. Time lapse between irradiation is a useful predictor of IQ. PMID- 11486701 TI - Assessment of proliferative activity in colorectal carcinomas by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). AB - The monoclonal antibody Ki-67 and the isospecific monoclonal antibody MIB-1 are routinely used in oncology to assess the proliferation index of tumor cells. A more objective and sensitive method is the determination of the of Ki-67 protein specific mRNA by quantitative reverse transcriptase-polymerase chain reaction (RT PCR). In 25 resected colorectal adenocarcinomas of different stages and grades we determined between 0.2 and 4.4 amol (10(-18) mol) Ki-67 protein-specific mRNA per microgram total RNA (median = 0.88 amol). The corresponding Ki-67 indices (expressing the percentage of Ki-67/MIB-I positive tumor cells) ranged from 41 to 81% (median = 61%). We found a good correlation between Ki-67 index and mRNA expression (r = 0.75), a significant correlation between both data and tumor stage (primary tumor, regional nodes, metastasis [pTNM] staging classification) (p < 0.001), but not between both data and tumor grade. Both Ki-67 indices (p = 0.05) and mRNA levels (p = 0.014) correlated significantly to the patients' survival. These results demonstrate that the Ki-67 protein-specific quantitative RT-PCR is a useful method for the characterization of tumor cell proliferation. PMID- 11486702 TI - Comparison of conventional dose and double dose carboplatin in patients receiving cyclophosphamide plus carboplatin for advanced ovarian carcinoma: a North Central Cancer Treatment Group Study. AB - Between March 1992 and November 1994, 91 patients with stage III and IV ovarian carcinoma were enrolled in a randomized comparative study of cyclophosphamide 600 mg/m2 plus carboplatin 300 mg/m2 vs. cyclophosphamide 600 mg/m2 plus carboplatin 600 mg/m2, each regimen given monthly for six cycles. Patients on the intensive regimen also received 10 micrograms/kg of granulocyte macrophage colony stimulating factor (GM-CSF) (molgramostim) daily for 14 days following each chemotherapy treatment. The study was closed prematurely because of very poor case accrual following the preliminary announcement (in May 1993) that paclitaxel appeared superior to cyclophosphamide in the platinum-based treatment of ovarian cancer. More than 4 years after our last case entry, we analyzed the survival results for the 44 eligible patients who received the conventional dose of carboplatin and the 43 eligible patients receiving our intensified dose of carboplatin. More than 90% of the treated patients receiving the conventional dose regimen received at least 75% of the planned doses at each of the six treatment intervals, whereas the percentage of treated patients able to receive at least 75% of the assigned intensive dose regimen had declined from 95% in cycle 2 to 53% by cycle 6. Furthermore, although 32 patients received all six planned cycles of treatment in the conventional regimen group, only 15 received all six cycles of the intensified regimen. Patients receiving the intensive regimen had more fever, dermatitis, lethargy, musculoskeletal pain, and pulmonary complications than did the conventional dose patients. Median survival times for the two treatment groups were very similar (38.5 and 38.1 months, respectively, for the conventional and intensive regimens), and we saw no evidence that the distribution of survival times differed between the treatment regimens (p = 0.95). PMID- 11486704 TI - Development of quantitative structure-activity relationship (QSAR) models for predicting risk of exposure from carcinogens in animals. AB - Quantitative structure-activity relationship (QSAR) models capable of predicting acute toxicity and carcinogen potency of polychlorinated dibenzo-p-dioxin (PCDD), polychlorinated hydrocarbons, and chlorinated insecticides have been formulated. Median lethal dose (LD50) for PCDD-exposed mice correlated negatively with polarity and positively with (H acceptor x 2 chi), whereas LD50 for PCDD-exposed guinea pigs correlated with (H acceptor x density). Both (H acceptor x 2 chi) and (H acceptor x density) exhibited parabolic relationship with log P (partition coefficient). Carcinogenic potency, determined from order of magnitude (OM) values, correlated negatively with log P and positively with (length x width). Thus, a hydrophobic mechanism plays a key role in the lethal effects of PCDD in mice, whereas both hydrophobic and electronic mechanisms are involved in the lethal effects of PCDD in guinea pigs. However, the molecule's lipophilicity, length, and width may play important roles in the carcinogenic effects of chlorinated compounds. PMID- 11486703 TI - Economic analysis of granulocyte colony stimulating factor as adjunct therapy for older patients with acute myelogenous leukemia (AML): estimates from a Southwest Oncology Group clinical trial. AB - Considerable morbidity, mortality, and economic costs result during remission induction therapy for elderly patients with acute myeloid leukemia (AML). In this study, the economic costs of adjunct granulocyte colony stimulating factor (G CSF) are estimated for AML patients > 55 years of age who received induction chemotherapy on a recently completed Southwest Oncology Group study (SWOG). Clinical data were based on Phase III trial information from 207 AML patients who were randomized to receive either placebo or G-CSF post-induction therapy. Analyses were conducted using a decision analytic model with the primary source of clinical event probabilities based on in-hospital care with or without an active infection requiring intravenous antibiotics. Estimates of average daily costs of care with and without an infection were imputed from a previously reported economic model of a similar population. When compared to AML patients who received placebo, patients who received G-CSF had significantly fewer days on intravenous antibiotics (median 22 vs. 26, p = 0.05), whereas overall duration of hospitalization did not differ (median 29 days). The median cost per day with an active infection that required intravenous antibiotics was estimated to be $1742, whereas the median cost per day without an active infection was estimated to be $1467. Overall, costs were $49,693 for the placebo group and $50,593 for the G CSF patients. G-CSF during induction chemotherapy for elderly patients with AML had some clinical benefits, but it did not reduce the duration of hospitalization, prolong survival, or reduce the overall cost of supportive care. Whether the benefits of G-CSF therapy justify its use in individual patients with acute leukemia for the present remains a matter of clinical judgment. PMID- 11486705 TI - The molecular biology of cervical cancer. AB - Cervical cancer remains a major worldwide health problem, especially in developing countries. Over the last few decades many advancements have been made in determining the molecular genetics of the development of cancer. This paper attempts to summarize the major disturbances in cellular function known to date to play a role in the development of cervical cancer. The role of human papillomavirus (HPV) infection in the development of cervical cancer is a major player in the genetic abnormalities described thus far. The effects of HPV E6 and E7 on important cell cycle genes are discussed. As oncogenes and tumor suppressor genes have been described in the different types of cancer, their possible role in cervical cancer has been investigated. The possible role of angiogenesis and angiogenic factors is described. Because of the importance of HPV infection in the development of cervical cancer, the role of the body's immune function in this cancer is also under study, and the results of these findings are summarized. Although a complete paradigm of the development of cervical cancer from normal cervical epithelium is not yet known, continued study in this area will hopefully lead to a defined progression of molecular and immunologic abnormalities that cause the disease. The goal would be to use this information to help prevent and/or treat cervical cancer in the future. PMID- 11486706 TI - Clues to the etiology of childhood brain cancer: N-nitroso compounds, polyomaviruses, and other factors of interest. PMID- 11486707 TI - Chemotherapy-induced menopause: a literature review. PMID- 11486708 TI - Selective estrogen receptor modulation: the search for an ideal hormonal therapy for breast cancer. AB - Female hormones, especially estrogens, play an important role in the pathogenesis of breast neoplasms and are a principal determinant of their biological behavior. Endocrine manipulation through medical or surgical means can often lead to objective shrinkage of breast tumors. Tamoxifen, a triphenylethylene estrogen receptor modulator, is currently the most widely used hormonal treatment for breast cancer. It has been conclusively demonstrated to reduce the risk of relapse following definitive local therapy (and systemic chemotherapy, when indicated) of invasive or noninvasive breast cancer. Recently, it has also been shown to reduce the incidence of breast cancer in healthy women who are at high risk of developing the disease. In addition, it can prevent osteoporosis and reduce the risk of fractures in postmenopausal women. However, its use is also complicated by an increased incidence of endometrial hyperplasia/carcinoma, venous thromboembolism, cataracts, and in some cases, emergence of tamoxifen dependent clones of breast cancer. These side effects (except cataracts) are believed to be related to estrogen-agonist effects of tamoxifen. Newer drugs, which are "pure antiestrogens" or inhibitors of estrogen biosynthesis, are devoid of such estrogen-agonist activity and may not have the liability of many of these side effects. However, these agents would also be expected to lack the potentially beneficial effects of tamoxifen on lipids and skeletal system. The ability of tamoxifen to act as an estrogen-agonist or estrogen-antagonist in a tissue-specific fashion has led to the concept of selective estrogen-receptor modulation. Selective estrogen receptor modulators (SERMs), which are devoid of estrogen-agonist effects on the uterus or breast cancer cells but retain potentially beneficial effects on bones and lipids, have been described as "ideal" SERMs. A number of such compounds are currently being tested. Raloxifene is already approved for prevention of osteoporosis and has potential efficacy for prevention and treatment of breast cancer. An analogue of raloxifene, LY353381, is currently in Phase II clinical trials for treatment of breast cancer, with promising early results. EM800 and CP336156 are other promising ideal SERMs in clinical trials. These compounds may provide better treatment and chemoprevention alternatives for breast cancer as compared to tamoxifen, aromatase inhibitors, and pure antiestrogens. In addition, they may also prove to be useful for the treatment and prevention of prostate cancer as well as for treating benign gynecological diseases such as fibroids and endometriosis. Future laboratory efforts should focus on further broadening the efficacy profile of SERMs (e.g., prevention of Alzheimer's disease and elevation of high-density lipoproteins to improve the likelihood of cardiovascular benefit) and narrowing their side-effect profile (e.g., risk of thromboembolism and hot flashes). PMID- 11486709 TI - Thrombopoietin. PMID- 11486710 TI - Personality-based typology of adolescent male sexual offenders: differences in recidivism rates, victim-selection characteristics, and personal victimization histories. AB - California Psychological Inventory scores from 112 adolescent male sexual offenders aged 12-19 (M = 15.59, SD = 1.46) were examined. A cluster analysis of factor-derived scores revealed four personality-based subgroups: Antisocial/Impulsive, Unusual/Isolated, Overcontrolled/Reserved, and Confident/Aggressive. Significant differences were observed between groups regarding history of physical abuse, parental marital status, residence of the offenders, and whether or not offenders received criminal charges for their index sexual assaults. Subgroup membership was unrelated to victim age, victim gender, and offenders' history of sexual victimization. Recidivism data (criminal charges) were collected for a period ranging from 2 to 10 years (M = 6.23, SD = 2.02). Offenders in the two more pathological groups (Antisocial/Impulsive and Unusual/Isolated) were most likely to be charged with a subsequent violent (sexual or nonsexual) or nonviolent offense. The four-group typology based solely on personality functioning is remarkably similar to that found by W. R. Smith, C. Monastersky, and R. M. Deisher in 1987 from their cluster analysis of MMPI scores. In addition to implications for risk prediction, the present typology is suggestive of differential etiological pathways and treatment needs. PMID- 11486711 TI - Multimodal assessment of dissociation in adolescents: inpatients and juvenile sex offenders. AB - Two samples of adolescents were studied to assess the validity of several measures of dissociation. The first sample included 70 males from a residential treatment program for adolescent sex offenders (ASO). The second were 47 psychiatric inpatients. The measures included the DSM-IV field trial questionnaire for Dissociative Disorders of Childhood, Child Dissociative Checklist, Adolescent-Dissociative Experiences Schedule, Trauma Symptom Checklist for Children, and the Dissociation Research scale. Ten of the 70 adolescent sex offender subjects (14.3%) were identified as meeting DSM-IV criteria for a Dissociation-spectrum disorder diagnosis and two (4.2%) of the psychiatric patients. Between-groups analyses with the ASO revealed significant differences on the Child Dissociative Checklist and the Dissociation Research Scale. Cumulative trauma did not differ significantly between the dissociative and nondissociative groups of ASO, but physical abuse was related to a diagnosis of dissociation in this sample. The dissociation measures were highly intercorrelated for both groups, and suggest that they measure a similar construct. PMID- 11486712 TI - Examining the substance use patterns and treatment needs of incarcerated sex offenders. AB - Using data from a Bureau of Justice Statistics' national prison inmate survey, this paper analyzes alcohol and drug use and abuse patterns among men incarcerated in state prison for sex crimes. Of the 13,986 inmates in the sample, 11.5% were incarcerated for a sex offense. Two thirds were substance-involved, meaning that they were under the influence of alcohol or drugs at the time of their crime, had committed a crime to get money for drugs, had histories of regular illegal drug use, had received treatment for alcoholism, or shared some combination of these characteristics. The level and type of substance-involvement was related to age and race, to history of victimization, and to victim characteristics. We discuss the implications of these findings for correctional program interventions, including assessing drug and alcohol problems, availability of substance abuse treatment for sex offenders, and the conjunction of such treatment with other programs. PMID- 11486713 TI - Brief report: dimensions of burnout in professionals working with sex offenders. AB - This report describes a brief questionnaire to investigate burnout potential in professionals working with sex offenders in clinical, forensic, and human services settings. The Professional Impact Questionnaire, a 31-item inventory with subscales assessing professional performance, coping strategies, and emotional reactions was tested initially with 17 clinicians who provide sex offender evaluations. In an altered format, the questionnaire was administered to 70 clinicians, jurists, and frontline caseworkers and their supervisors. The satisfactory psychometric properties of the original questionnaire were retained in the modified version with this new sample of respondents. Caseworkers reported significantly greater emotional distress than attorneys and judges. For all participants, negative emotional reactions were associated with a sense of impaired work performance. Using positive coping strategies was correlated with reporting relatively few adverse effects on professional functioning. PMID- 11486714 TI - Treatment of sexual offenders who are in categorical denial: a pilot project. AB - This paper describes an approach to treatment for sexual offenders who are in categorical denial. Other efforts to have them, at least partially, admit responsibility had failed and they were to be released from prison without any treatment. Evidence that suggests denial is not predictive of risk and that treatment may reduce the risk of these offenders is reviewed. Essentially, this paper offers a possible approach to dealing with these intractable deniers which, it is suggested, is better than not trying to modify their risk, and that may prove to be effective. PMID- 11486715 TI - A revision of the Abel and Becker Cognition Scale for intellectually disabled sexual offenders. AB - The Abel and Becker Cognition Scale (ABCS) measures cognitive distortions supportive of sexually assaultive behavior by child molesters. Research has shown that ABCS items may be too complex to be comprehended by offenders with intellectual disabilities. A modification of the ABCS to increase its readability may be one way to facilitate the valid assessment of the cognitive distortions of intellectually disabled offenders. In addition, a dichotomous scoring system was found to be helpful in the reduction of extremity bias by such offenders. The revised items, explanation of the scoring system, and supporting data are available from the first author. PMID- 11486716 TI - In defense of utilizing the construct of cognitive distortions: a commentary. PMID- 11486717 TI - Women and HIV: across class, cultures, and countries. PMID- 11486718 TI - Negotiation of HIV preventive behaviors in divorced and separated women reentering the sexual arena. AB - The purpose of this study was to understand divorced and separated women's experience of negotiating human immunodeficiency virus (HIV) preventive behaviors when resuming sexual activity following long-term monogamous relationships. A little-studied but large group, these women are at risk for sexually transmitted diseases, have low rates of protective behaviors, and have not been targeted for prevention education. Seven women entering a sexual milieu altered by the presence of HIV after at least a 10-year period of monogamy comprised the sample. A descriptive phenomenologic methodology incorporating Colaizzi's method for data analysis revealed four themes: (a) keeping the magic, (b) Am I an adolescent or an adult? (c) holding on to self, and (d) gender perspectives. These women's words help to illuminate their issues regarding HIV prevention, guiding further nursing research and HIV preventive educational interventions. PMID- 11486719 TI - Physical and mental health in African American mothers with HIV. AB - This article examines the physical and mental health of African American mothers during a 2-year period following the birth of an infant seropositive for human immunodeficiency virus (HIV). Participants were 34 African American mothers enrolled when the infants were approximately 3 months of age and reinterviewed when the infants were 6, 12, 18, and 24 months. Three self-report questionnaires were used to assess physical health (perception of health, activity limitation, and physical symptoms) and mental health (depressive symptoms and stigma). Health symptoms most often reported were infections, problems thinking and remembering, low energy, and gynecologic problems. Moderate levels of perceived stigma were reported. Depressive symptoms were high; a large number of women at each data point had depressive symptom scores above the cutoff, indicating risk for depression. There were significant correlations between depressive symptoms and health, suggesting a link between mental and physical health. These findings have significance for the health of the mother and the parenting of her infant. Attention should be paid to the mental and physical health of mothers with HIV, especially during the first 2 years after the birth of a child. PMID- 11486720 TI - Evaluation of two self-care treatments for prevention of vaginal candidiasis in women with HIV. AB - Vaginal candidiasis (VC) is a common concern for women living with HIV infection. The authors evaluated the effectiveness of two self-care approaches to prophylaxis of VC among HIV-infected women, weekly intravaginal application of Lactobacillus acidophilus or weekly intravaginal application of clotrimazole tablets, in a randomized, double-blind, placebo-controlled trial. VC was defined as a vaginal swab positive for Candida species in the presence of signs/symptoms of vaginitis and the absence of a diagnosis of Trichomonas vaginalis or bacterial vaginosis. Thirty-four episodes of VC occurred among 164 women followed for a median of 21 months. The relative risk of experiencing an episode of VC was 0.4 (95% CI = 0.2, 0.9) in the clotrimazole arm and 0.5 (95% CI = 0.2, 1.1) in the Lactobacillus acidophilus arm. The estimated median time to first episode VC was longer for clotrimazole (p = .03, log rank test) and Lactobacillus acidophilus (p = .09, log rank test) compared with placebo. Vaginal yeast infections can be prevented with local therapy. Education about self-care for prophylaxis of VC should be offered to HIV-infected women. PMID- 11486721 TI - HIV treatment adherence in women living with HIV/AIDS: research based on the Information-Motivation-Behavioral Skills model of health behavior. AB - Close and consistent adherence to anti-HIV medication regimens is necessary to achieve the maximum benefit of these potentially effective treatments. The authors examined cognitive and behavioral factors associated with HIV treatment adherence in a convenience sample of 112 women, 72 of whom were currently taking HIV treatments at the time of the study. Women completed confidential surveys and interviews to assess HIV-related health status, treatment regimens, and cognitive behavioral characteristics derived from the Information-Motivation-Behavioral Skills model of health promotion behaviors. Results showed that women who had missed at least one dose of their HIV medications in the past week reported lower intentions (motivation) to remain adherent and lower adherence self-efficacy (skills). Structural equation modeling showed that motivational and skills building factors significantly predicted the number of medication doses missed. However, treatment-related information did not predict treatment adherence. In addition, women who had missed a dose of medication in the past week were more likely to have ever used devices and strategies to remind them of doses, but were no more likely to currently use such strategies. Interventions that enhance treatment adherence motivation and build adherence skills may help improve HIV treatment adherence in women receiving anti-HIV therapies. PMID- 11486722 TI - Social interactions, perceived support, and level of distress in HIV-positive women. AB - Although powerful pharmacological therapies are helping women with HIV infection live longer, women continue to experience the stressors of chronic illness. This study used a person-environment systems framework to describe social interactions, perceived social support, and psychological distress in HIV positive women. A convenience sample of 104 HIV-positive women living in the San Francisco Bay Area completed a questionnaire on social interactions, perceived social support, and distress. Women reported limited social interactions with family and friends and a low level of perceived social support. Social support and level of distress did not differ by ethnicity. Limited perceived social support was a significant predictor of distress in this sample of women. Supportive interactions from health care providers can be useful in mediating the relationship between the stressor of HIV disease and distress in HIV-positive women. Community-based nurses can enhance HIV-positive women's support network by providing positive and supportive interactions as they intervene with women in symptom management, case management, and other health care services. PMID- 11486723 TI - Content validity of a Scale to measure silencing and affectivity among women living with HIV/AIDS. AB - This study measured quantitatively and explored qualitatively silencing behaviors and affectivity (mood) in women living with HIV/AIDS and confirmed the validity of the Silencing the Self Scale and the Positive and Negative Affect Scale. Silencing behaviors are interpersonal communication styles that suppress personal needs and feelings to preserve relationships with others. Silencing behaviors serve as protective strategies that allow one to divorce oneself from an overbearing culture. Affectivity is a way of measuring one's personal mood state by a positive to negative continuum. The results indicate that the women silenced themselves profoundly, especially when it came to putting the needs of their children or dependents before their own. The women also had high levels of negative affectivity. The research findings from this study extend nursing knowledge by addressing the unique social processes of women living with HIV/AIDS within health care service structures and significant social groups. Further exploration of "silencing" as a phenomenon of this group through measurement and experience will help define specific interventions that are meaningful to and for women living with HIV/AIDS. PMID- 11486724 TI - Focus group methodology: when and why? PMID- 11486725 TI - Serono initiates clinical trial of Serostim for AIDS-related metabolic complication. PMID- 11486726 TI - Federal Drug Administration panel recommends approval of Cancidas, Merck's investigational antifungal drug. PMID- 11486727 TI - HIV resistance survey shows majority of physicians choose single protease inhibitor as first-line combination therapy. PMID- 11486728 TI - HIV drug resistance fact sheet. PMID- 11486729 TI - Stem cell dogmas in the genomics era. AB - Recently, much excitement has been generated by strong suggestions that stem cells isolated from diverse somatic tissues may have a previously unsuspected degree of developmental or differentiation plasticity. For example, a hematopoietic stem cell may be capable of producing mature liver cells, muscle tissue or even neurons. Similarly, central nervous system stem cells or muscle stem cells may be capable of producing mature blood cell populations. These observations have called into question several fundamental dogmas of developmental biology. In addition, these observations offer extraordinary promise in the clinical setting. It is of paramount importance to rigorously assess the suggested plasticity phenomena using precise clonal analysis. In order to explore the plasticity phenomena in more direct ways, it is necessary to develop in vitro systems where such behavior can be recapitulated in a well defined setting. Finally, stem cell plasticity will be governed, at least in part, by cell-autonomous mechanisms: that is, those mediated by the panel of gene products expressed in stem cells. Therefore, it is necessary to identify the complete gene expression profile that defines the stem cell. PMID- 11486730 TI - Adult stem cell plasticity and methods of detection. AB - The ability to selectively produce one or more differentiated cell types at will from totipotent stem cells would be of profound clinical importance, as it would enable the specific replacement of damaged/dysfunctional cell types within the body, potentially curing numerous diseases. Until recently, it was thought that the only cells that possessed sufficient immaturity to be capable of giving rise to more than one tissue type in vitro and in vivo were the embryonic stem cells. However, recent studies have now provided compelling evidence that the adult bone marrow, brain and skeletal muscle contain stem cells that possess the remarkable ability to trans-differentiate and give rise to progeny of alternate embryologic derivations. These recent findings have shattered the existing dogma that the stages of embryologic development are irreversible. In this review, we present a brief summary of the most significant findings in the field of stem cell plasticity, emphasizing studies involving the hematopoietic system, discussing the models used thus far, and finishing with our findings on human stem cell plasticity using the fetal sheep model. PMID- 11486731 TI - Purification and functional assay of pluripotent hematopoietic stem cells. AB - Hematolymphopoietic stem cells (HSC) have the capacity for extensive self-renewal and pluripotent myelolymphoid differentiation. Recent studies have emphasized the heterogeneity of human HSC subsets in terms of proliferative and self-renewal capacity. In the NOD-SCID (nonobese diabetic-severe combined immunodeficient) mouse xenograft assay, most CD34+38- stem cell clones proliferate at early times, but then disappear, whereas only few clones persist: possibly, the latter ones consist of long-term engrafting CD34+38- HSC expressing the KDR receptor (i.e. the vascular endothelial growth factor receptor II). In this regard, isolation of the small KDR+ subset from the CD34+ hematopoietic progenitors (and possibly from the CD34-lin- population) may provide a novel and effective approach for the purification of long-term proliferating HSC. More importantly, KDR+ HSC isolation will pave the way to cellular/molecular characterization and improved functional manipulation of HSC/HSC subsets, as well as to innovative approaches for HSC clinical utilization, specifically transplantation, transfusion medicine and gene therapy. PMID- 11486732 TI - Normal and leukemic CD34-negative human hematopoietic stem cells. AB - Considerable progress has been made in recent years in purifying human and murine hematopoietic stem cells. The essential marker identified is the sialomucin CD34, which is expressed on primitive cells and downregulated as they differentiate into more abundant mature cells. CD34 is not unique to stem cells, however, as it is also expressed on clonogenic progenitors and some endothelial cells. Nevertheless, all clinical and experimental protocols are targeted to CD34+ cells enriched by a variety of selection methods. Recent studies in both the murine and human systems have indicated that some stem cells capable of multilineage repopulation do not express detectable levels of cell surface CD34. These studies challenge the dogma that all human repopulating cells are found in the CD34+ subset. However, the precise relationship between CD34- and CD34+ stem cells is still not well understood. In this review, the results on the discovery of the CD34- repopulating cell are summarized and the impacts this discovery may have, both clinically and in our understanding of the organization of the human hematopoietic system, are examined. PMID- 11486733 TI - Role of surfactant protein A (SP-A)/lipid interactions for SP-A functions in the lung. AB - Surfactant protein A (SP-A), an oligomeric glycoprotein, is a member of a group of proteins named collectins that contain collagen-like and Ca(2+)-dependent carbohydrate recognition domains. SP-A interacts with a broad range of amphipathic lipids (glycerophospholipids, sphingophospholipids, glycosphingolipids, lipid A, and lipoglycans) that are present in surfactant or microbial membranes. This review summarizes SP-A/lipid interaction studies regarding the lipid system used (i.e., phospholipid vesicles, phospholipid monolayers, and lipids immobilized on silica or adsorbed on a solid support). The effect of calcium, ionic strength, and pH on the binding of SP-A to lipids and the subsequent lipid aggregation process is discussed. Current evidence suggests that hydrophobic-binding forces are involved in the peripherical association of SP-A to membranes. It is also proposed that fluid and liquid-ordered phase coexistence in surfactant membranes might favor partition of SP-A into those membranes. The binding of SP-A to surfactant membranes containing hydrophobic surfactant peptides makes possible the formation of a membrane reservoir in the alveolar fluid that is protected by SP-A against inactivation and improves the rate of surfactant film formation. In addition, the interaction of SP-A with membranes might enhance the affinity of SP-A for terminal carbohydrates of glycolipids or glycoproteins on the surface of invading microorganisms. PMID- 11486734 TI - Surfactant regulation of host defense function in the lung: a question of balance. AB - Although the lung is protected by classic innate and adaptive immune mechanisms, another unique local immunoregulatory system involving pulmonary surfactant is described in this review. Normal surfactant inhibits many immune cell functions including proliferation resulting from various stimuli and production of reactive oxidants, inflammatory mediators, and some cell surface markers. The predominant surfactant lipids appear to be responsible for these suppressive effects. Conversely, surfactant proteins SP-A and SP-D stimulate many aspects of immune cell behavior. These proteins are collagenous lectins or collectins that bind to glycoconjugates on many pathogens, enhancing phagocytosis and killing in some cases. SP-A and SP-D stimulate chemotaxis and reactive oxidant generation, particularly in macrophages, although other cells are probably affected as well. In some cases, SP-A also stimulates the expression of cell surface markers and is involved in the stimulation of inflammatory mediators. Under normal conditions, the inhibitory effects of the lipid prevail, but the collectins may provide focal activation and stimulate immune cells at sites where they are needed. However, in some types of lung disease or after certain insults or exposures, the balance between these inhibitory and stimulatory influences may be disrupted and result in inflammatory injury. PMID- 11486735 TI - Functional mapping of surfactant protein A. AB - Surfactant protein A (SP-A) is a highly ordered, oligomeric glycoprotein that is secreted into the airspaces of the lung by alveolar type II cells and Clara cells of the pulmonary epithelium. Although research has shown that SP-A is both a calcium-dependent phospholipid-binding protein that affects surfactant structure and function and a lectin that opsonizes diverse microbial species, our understanding of the physiologically relevant roles of SP-A in the lung remains incomplete. My review focuses on the putative biological functions of SP-A that are supported by experiments in mammals and on the structural basis of SP-A function. PMID- 11486736 TI - Localization and functions of SP-A and SP-D at mucosal surfaces. AB - Pulmonary surfactant protein A (SP-A) and D (SP-D), members of the collectin family, are implicated in innate host defense of the lung. Collectins consist of a collagen-like domain and a carbohydrate recognition domain. SP-A and SP-D recognize and interact with glycoconjugates on the surface of microorganisms. They protect the lung by interacting with a wide variety of potential pathogens, including viruses, bacteria, and fungi. This may result in enhanced killing and/or clearance by phagocytes. Although most extensively studied in the lung, both SP-A and SP-D, or proteins closely resembling SP-A and SP-D, are found in a number of other sites in the body and therefore may play a protective role at other sites than the lung. SP-A and SP-D protein and/or mRNA have been detected at various sites of the body: the respiratory tract, the gastrointestinal tract, the middle ear, and in the peritoneal cavity. The presence of SP-A and SP-D at these mucosal surfaces, in close contact with numerous potentially harmful microorganisms, supports a role for these "lung"-collectins in innate mucosal defense. SP-A and SP-D may be important molecules in a threefold innate defense, particularly in the neonatal period between maternally acquired immunity and a fully developed adaptive immune system; the time interval between first exposure to a pathogen and generation of specific antibodies; and states of impaired immune function. PMID- 11486738 TI - Pathology quiz: small cell osteosarcoma. AB - A 15-year-old black male presented with shortness of breath, leg weakness, and pain in his back and rib cage. Four years previously he had noticed a lump in his upper back and complained of pain when playing basketball, especially on contact to that area. Recently, the pain had become more constant and increased in intensity. This was associated with loss of control in his legs, weakness, and paraesthesia. General physical examination revealed a palpable mass in the right midline upper back. Laboratory results were within normal limits. Radiographic scans demonstrated a destructive soft tissue mass at T6 vertebral body with scattered stippled calcification (Figure 1). The patient underwent a biopsy followed by excision of the mass (Figure 2) and decompressive laminectomy with reconstruction. PMID- 11486737 TI - Pathology teach and tell: vaginal mesonephric papilloma. PMID- 11486739 TI - Structure, function, and expression of pulmonary surfactant proteins; considerations for use in artificial surfactants. Part I. Introduction. PMID- 11486740 TI - [Morphological characteristics of peripheral blood erythrocytes in patients with disseminated atherosclerosis]. AB - Studies of surface architectonics of erythrocytes in 20 patients with atherosclerotic involvement of the lower limb arteries by scanning electron microscopy showed clear-cut redistribution of the erythrocyte population, which depended on the cell morphology, with a tendency to a decrease in the count of discocytes and an increase in the counts of echinocytes, spherocytes, stomatocytes, and degeneratively changed erythrocytes. PMID- 11486741 TI - [Isolation of blood eosinophils in multi-step density gradient]. AB - The density and morphology of blood eosinophils from 60 children with atopic dermatitis were investigated. The proportion of hypodense eosinophils is higher during the acute stage of disease than during remission (59.78 +/- 0.20% vs. 21.40 +/- 0.04%). PMID- 11486742 TI - [Indices of proteolysis in evaluation of resorption in bone tumors]. AB - Activities of acid and alkaline phosphatases, collagenase, cathepsin D, trypsin like proteinases, alpha(1)-proteinase inhibitor (alpha(1)-PI), alpha(2) macroglobulin (alpha(2)-MG) were measured in blood plasma and tumor tissue of patients with giant-cell tumor of the bone (GCTB) and bone chondrosarcoma. These tumors differed by enzymatic activities. GCTB is characterized by increased activity of alkaline phosphatase, while in chondrosarcoma tissue the activities of collagenase and cathepsin D were the highest. Activities of acid phosphatase, collagenase, trypsin-like proteinases were increased in the plasma of patients with both tumors; alpha(1)-PI/alpha(2)-MG ratio was increased. Bone resorption parameters correlated with proteolysis inhibitors. Activities of collagenase and acid phosphatase were increased in tumor tissue and plasma in the presence of low activities of alpha(2)-MG and increased alpha(1)-PI/alpha(2)-MG index, which seems to require special attention during the postoperative period. PMID- 11486743 TI - [Very low density and low density lipoproteins: function, transport of fatty acids and diagnostic role (lecture)]. PMID- 11486744 TI - [Quantitative evaluation of the activity of nuclear Ca2+/Mg2+-dependent endonucleases in hyperplasia and cancer of the endometrium]. AB - Endometrial carcinoma is the most incident cancer of human reproductive system. There are unequivocal evidences of relationship between complex and atypical hyperplasia and development of cancer. Apoptosis plays a significant role in the maintenance of equilibrium between cell death and proliferation and contributes to prevention of tumorigenesis. Internucleosomal DNA fragmentation known as one of the most important criteria of apoptosis cannot be used for evaluating the risk of cancer development because it reflects the current level of apoptosis but is useless for evaluating the real limits of apoptosis intensity in certain types of tissue. For estimating the possibility of apoptosis development in endometrial tissues, a new method of quantitation of nuclear Ca(2+)/Mg(2+)-dependent endonuclease (NCME) activity has been developed. Fifteen samples of normal endometrial tissues at middle proliferative, secretory, premenstrual, and menstrual phases, 43 samples of hyperplastic endometrial tissues, 13 samples of endometrial polyps, and 17 samples of endometrial adenocarcinoma were collected by diagnostic curettage of the uterine cavity and by hysterectomy (carcinomas). The material was examined by 1) TUNEL method and 2) agarose gel electrophoresis of DNA cleaved by nuclear CME in isolated cell nuclei in the presence of Ca(2+) and Mg(2+) ions, followed by quantitation of CME activity. The activity of NCME was found to decrease from normal endometrium (1.1 +/- 0.12 U, without significant changes throughout the menstrual cycle) through polyps (0.9 +/- 0.15 U), cystic hyperplasia (0.45 +/- 0.06 U, p < 0.01), and adenomatous hyperplasia (0.32 +/- 0.08 U, p < 0.01) to adenocarcinoma (0.37 +/- 0.11 U, p < 0.01 for well differentiated, 0.16 +/- 0.08 U, p < 0.01 for moderately differentiated, and 0.03 +/- 0.02 U, p < 0.01 for poorly differentiated samples). The TUNEL-specific staining pattern in normal endometrium varied in a wide range during the menstrual cycle (from poorly stained individual cells in the proliferative phase to intensely stained cell clusters in the premenstrual phase). At the same time the difference between the normal endometrium in the proliferative phase and pathologically changed endometrium (hyperplasia or cancer) could not be detected by the TUNEL technique. Hence, TUNEL is useless for predicting cancer development in hyperplasia and precancer. By contrast, evaluation of NCME activity helps detect the early disorders in the proliferative processes coursing in endometrial tissues and thus prevent tumor development. PMID- 11486746 TI - [Clinico-laboratory parallels during celiac disease in children]. AB - Forty-five patients with celiac disease and 73 with the celiakia syndrome were observed. The clinical picture was identical and was characterized by disordered intestinal absorption. Depression of T and B lymphocytes was typical of both forms of the disease. High values of antigliadin IgA and IgG correlated with the severity of atrophic processes in the small intestinal mucosa. Intestinal microflora was characterized by a decrease in the main defense flora and growth of hemolytical and lactonegative enterobacteria and Proteus. Hypocholesterolemia was characteristic of congenital and acquired celiakia. Hemoglobin and albumin levels were in direct correlation while growth deficiency and increment of glycemia were in inverse correlation in patients with celiac disease. The celiakia syndrome was characterized by an inverse correlation between the number of defecations and content of full-value E. coli, body weight deficit, and glucose tolerance test. PMID- 11486745 TI - [Optimization of the anti-Helicobacter therapy of peptic ulcer on the basis of modern methods of pharmacokinetic analysis]. PMID- 11486747 TI - [Lipid peroxidation and antioxidant system in inhalation injury]. AB - Thirty-five patients (aged 45 +/- 2.0 years) with isolated inhalation injury were examined on days 1, 3, and 7 after the injury, Serum concentrations of lipid peroxidation (LPO) products (dienic conjugates, malonic dialdehyde, and degree of lipid oxidation) were increased in all terms of investigation. Studies of the antioxidant system (AOS) showed a decrease in endogenous alpha-tocopherol during all terms, while ceruloplasmin content gradually increased. LPO/AOS imbalance was the most expressed on day 7 after the injury, which indicates intensification of LPO processes in the victims. The results pathogenetically validate the use of antioxidants in combined therapy of patients with inhalation injuries. LPO/AOS imbalance should be evaluated in these patients. PMID- 11486748 TI - [Relationship between the quality of results and the use of analytical systems in routine clinical chemistry]. PMID- 11486749 TI - An unprovoked tonic-clinic seizure should generally not be treated. PMID- 11486750 TI - Set size and repetition matter: comment on Caramazza and Costa (2000). AB - Caramazza and Costa, 2000 (Cognition 75, B51--B64) report three picture-word interference experiments testing the response set mechanism of the WEAVER+ + model of spoken word production. They argue that their findings are problematic for WEAVER+ + and that the model's architecture needs to be changed. I show that there is no need to fundamentally modify the model. Instead, the findings of Caramazza and Costa, and all previous findings, are explained by assuming that only a limited number of responses can be kept in short-term memory and that memory improves with response repetition. PMID- 11486751 TI - Retraction notice for the brief article "Goal attribution in chimpanzees" by Claudia Uller and Shaun Nichols. Published in Cognition volume 76 (2000) pp. B27 B34. PMID- 11486752 TI - [Is the dental wholesale trade in Netherlands dependent on the Internet?]. PMID- 11486753 TI - Thromboembolism in pregnancy: a new temporary caval filter. AB - BACKGROUND: Our aim was to check out the advantages of temporary caval filtration in pregnancy with a retrievable filter: Tempofilter (tCF). METHODS: During a period of 2 years we implanted tCF in four patients at the third pregnancy trimester with serious acute deep venous thrombosis (DVT) of the lower limbs for prophylaxis of pulmonary thromboembolic disease (TED) in the pre-partum, during Caesarian section (CS), and in the post-partum period. tCF has been placed percutaneously in local anaesthesia using the Seldinger technique and under image intensifier control. All procedures were performed in a polyvalent ICU. RESULTS: tCF positioning was always easy. No complications linked to the adopted technique occurred. No incidence of pulmonary embolism was reported (PE). The permanence period of tCF in situ was 14 to 29 days (mean+/-SD: 21+/-6). The CS always took place without complications either for the patient or for the foetus. CONCLUSIONS: The combined use of tCF and an anti-coagulating therapy resulted in an effective prophylaxis of TED for the pregnant woman without any interferences with CS and with no consequence as for permanent endovascular prosthesis for the young patient herself. PMID- 11486754 TI - Alternative technique for subclavian vein cannulation. AB - BACKGROUND: Subclavian vein catheterism, a commonly used methodology in ICU, is not devoid of complications, even some serious ones (Pneumothorax 0.5%). METHODS: The authors propose an alternative technique based on the use of an introducer needle bent on its own axis to form an arch with its concavity turned towards the Quincke point. On searching the blood vessel, once the subclavian plane has been reached, the needle advancement towards the jugular fosse happens by following the needle curvature. In this way the operator can keep the needle on a plane parallel to the cutaneous one, and all the complications due to involuntary exceeding of the costal plane are avoided. The response of this new technique in 110 patients (study group) is compared through a retrospective study, to 100 patients with whom the standard technique has been used (control group). RESULTS: The study group has presented a significant reduction both of total complications (p<0.05) and of total failures (p< 0.01). CONCLUSIONS: Such results, although pertinent to a still limited study, testify to an improvement in terms of security and efficacy, with the new alternative technique without an increase in costs. PMID- 11486756 TI - Ezzo et al.: Is acupuncture effective for the treatment of chronic pain? A systematic review, PAIN 86(2000)217-225. PMID- 11486755 TI - Re.: A pair of publications. PMID- 11486760 TI - Re.: Clinical, radiological and pathological findings in an autosomal dominant leukodystrophy. PMID- 11486761 TI - The UK burn unit experience of the treatment of non-burn conditions. PMID- 11486762 TI - Recent references. PMID- 11486763 TI - Achieving compliance for biologics. PMID- 11486764 TI - Do'fail-clean' ecostrategies deliver 'fail-safe' outcomes? PMID- 11486765 TI - In response to the February 2001 article entitled "Improving access to mammography through community-based influenza clinics: a quasi-experimental study". What exactly is a quasi-experimental study? PMID- 11486766 TI - Effect of different methods of administration of ivermectin on its efficacy against the shedding of gastrointestinal nematode eggs by gazelles. AB - Seven different methods of administering ivermectin to gazelles were compared: subcutaneous injection, direct oral administration, administration in individual feeds, administration in a herd feed, direct oral administration of a second ivermectin formulation, administration in individual water supplies, and administration in the herd's water supply. The first five treatments were effective, as monitored by faecal egg count reduction tests, and administration in individual feeds or in a herd feed avoided the need to capture the animals, with the attendant risk of mortality. Of the factors associated with the recipients (species, sex, age and inbreeding coefficient) age was the only significant factor for the efficacy of the treatment. Oral or subcutaneous, individual or collective, and direct or indirect administrations were equally satisfactory for the treatment of all the parasite groups studied. Only when parasitic problems were due to Nematodirus species did direct administration to individual animals appear to be preferable. PMID- 11486767 TI - Treatment of irreducible caecocolic intussusception in horses by jejuno(ileo)colostomy. AB - Six of eight horses with caecocolic intussusception were treated successfully by jejuno- or ileocolostomy. The other two horses were euthanased during surgery. Four of the six horses survived long term, but two died within two-and-a-half months, of problems related to the surgery. Compared with other techniques for treating caecocolic intussusception, jejuno- or ileocolostomy reduces surgical time and decreases the risk of abdominal contamination. PMID- 11486768 TI - Isolation of Borrelia afzelii from a dog. PMID- 11486769 TI - Veterinary surgeon to retrain in surgery and anaesthesia after admitting misconduct. PMID- 11486770 TI - Aerophaia and gastric dilation following tiletamine/zolazepam anaesthesia in a dog. PMID- 11486771 TI - Immunohistochemical detection of ophidian paramyxovirus in snakes in the Canary Islands. PMID- 11486772 TI - Dermatosis in talapoin monkeys (Miopithecus talapoin) with response to zinc and animal protein. PMID- 11486773 TI - Body temperature patterns during natural fevers in a herd of free-ranging impala (Aepyceros melampus). PMID- 11486774 TI - Anaesthesia of a pregnant alpaca (Lama pacos). PMID- 11486775 TI - Preparing for epizootics. PMID- 11486776 TI - Idiopathic mouth ulcers in sheep. PMID- 11486777 TI - Role of the RCVS. PMID- 11486778 TI - PDSA review. PMID- 11486780 TI - Ear tip necrosis in standard poodles. PMID- 11486781 TI - Piroplasmosis in southern France. PMID- 11486782 TI - Treatment of fungal infections in dogs and cats. PMID- 11486783 TI - Effects of stone chewing by outdoor sows on their teeth and stomachs. AB - Stone chewing is a common behaviour in outdoor sows, but its effects on their teeth and stomachs have not been investigated. Tooth wear and damage was assessed by examining the heads of 58 sows culled from outdoor units and 23 culled from indoor units. Tooth damage was found in 28 per cent of outdoor and 30 per cent of the indoor sows, and tooth wear affected 88 per cent of the outdoor and 91 per cent of the indoor sows. The outdoor sows were more prone to wear on the lower molars and premolars, a pattern of wear associated with stone chewing. The stomachs of 152 outdoor sows and 47 indoor sows were examined. Stones were found in 59 of the outdoor sows but in none of those kept indoors. There was no evidence that the presence of stones damaged the stomach, or that stone chewing affected the health of the sows. The teeth of the outdoor sows were worn, but probably not sufficiently to affect their ability to eat during their relatively short productive lives. PMID- 11486784 TI - [Analysis of national maternal mortality rate surveillance. Maternal Death Investigation Cooperative Group of China]. AB - OBJECTIVE: To analyze the results of maternal mortality rate (MMR) surveillance in 247 monitoring areas. METHODS: Based on the demographic characteristics of different areas, stratum sampling method was used, and 247 monitoring points from 30 provinces, autonomous regions and cities covered about 100 million population were selected in China in 1989-1995. RESULTS: In the 7 years of monitoring there were 8,709,220 cases of live births with 5,984 cases of maternal deaths in these monitoring points. The average MMR was 77.4/100,000 with 45.5/100,000 in urban areas and 95.4/100,000 in rural areas. There were significant differences of MMR from area to area from 26.0/100,000 to 308.0/100,000. The causes of maternal deaths in succession were obstetric hemorrhage, pregnancy induced hypertension, heart diseases, amniotic fluid embolism, puerperal infection and hepatic diseases, and these 6 causes constituted 81.6% of the total. According to the audit of mortality cases demanded by WHO, the results showed that there was 73.0% avoidable deaths and 27.0% unavoidable deaths. The main problems were: 49.7% among the medical and health care system, chiefly lack of knowledge and skill of the medical personnels; 44.5% of the pregnant women and their family members lack the basic health knowledge. CONCLUSION: To improve the living condition of both mother and children, to increase hospital delivery rate, training medical personnel's skill, to provide better maternal health care, and to strengthen health education are effective measures to decrease MMR. PMID- 11486785 TI - How reliable is the ultrasound densitometer for community screening to diagnose osteoporosis in spine, femur, and forearm? AB - Dual X-ray absorptiometry (DXA) is the most widely used noninvasive, quantitative diagnostic method of measuring bone density, but it exhibits several limitations for the purpose of general screening and community-based study. Thus, we evaluated the feasibility of the ultrasound densitometer SoundScan Compact for screening the osteoporosis community. We compared the reliability of the SoundScan Compact's measurement to predict both osteopenia and osteoporosis at the spine, femur, and forearm measured by the XR-36. World Health Organisation criteria and Korean reference were used for the diagnosis of osteopenia and osteoporosis. Of the 1827 subjects screened in the community, we randomly selected and invited 106 subjects to our university hospital center for clinical epidemiology for further evaluation of their bone health using DXA. The site specific relationships among speed of sound and bone mineral density at L2-L4 spine, femoral neck, trochanter, Ward's triangle, distal radius + ulna, proximal radius + ulna, and proximal radius were highly correlated with r values of 0.633, 0.632, 0.674, 0.632, 0.668, 0.78, and 0.777 respectively (all p < 0.0001). Of the 42 subjects defined as normal in the spine by SoundScan Compact's, 27 (64.3%) were also in agreement as the normal by the XR-36. However, an agreement of 43.5% (10/23) in osteopenia and 53.7% (22/41) in osteoporosis was less sensitive. Reliability tested by kappa statistics (kappa = 0.35; t = 5.3; p < 0.0001) revealed that the degree of agreement was not coincidental. On the femur, 54.8% (23/42) agreed as normal, 73.9% (17/23) as osteopenia, and 39% (16/41) as osteoporosis (kappa = 0.33; t = 5.5; p < 0.0001). Higher reliability was found in the forearm, with a kappa value of 0.48, t = 7.2, and p < 0.0001. A magnitude of agreement was also greater in the forearm, with 83.3% (35/42) agreement in normal, 52.2% (12/23) in osteopenia, and 64% (16/25) in osteoporosis. Although the SoundScan Compact's measures different bone properties and measurements are taken at different sites, it is a very useful instrument with a high correlation and high degree of variation and reliability to predict bone health at other sites such as the spine, femur, and forearm. The ultrasound densitometer such as SoundScan Compact's merits its use for community screening and in the primary care setting to detect osteopenia and osteoporosis for the purpose of early detection and prevention. PMID- 11486786 TI - Building knowledge in nursing and midwifery--European Academy of Nursing Science. PMID- 11486787 TI - Puberty and prognosis of asthma and bronchial hyper-reactivity. AB - It is a commonly held view that pediatric asthma frequently abates during puberty. However, little data are available that associate the stage of puberty with the prognosis of asthma and bronchial hyper-reactivity (BHR). In this study, 155 children with active asthma at 10 years of age (60 girls [38.70%], 95 boys [61.3%]) were followed-up until they reached 14 years of age. The stage of puberty was assessed by parental questionnaire; in addition, serum 3-alpha androstanediolglucuronide, as an endocrinological marker for peripheral androgen status, was measured in 107 subjects. Persistence of asthma was determined via questionnaire, lung function testing, and bronchial provocation (hyperventilation of cold, dry air). At 14 years of age, 73.3% of girls were reported to have had menarche and 40.8% of boys a voice change, and only 35.5% of the subjects had experienced acute asthma symptoms during the last 12 months, with an almost unchanged gender ratio (19 girls [34.5%], 36 boys [65.5%]) vs. that recorded at 10 years of age. The level of androstanediolglucuronide was higher in the children who reported puberty (mean+/-SD): 3.03+/-2.13 nmol/l vs. 1.89+/-1.26 nmol/l, p = 0.003. No statistically significant relationship was found between the reported signs of late puberty and loss of asthma or BHR. Likewise, no significant association was found between asthma persistence and the level of androstanediolglucuronide (2.39+/-1.75 nmol/l vs. 2.44+/-1.82 nmol/l, p = 0.84), or BHR and the level of androstanediolglucuronide (3.02+/-1.97 nmol/l vs. 2.28+/ 1.67 nmol/l, p = 0.13), at 14 years of age, in girls or boys. At 14 years of age, no change in the gender ratio of children with active asthma had occurred. These results may indicate that the change in gender predominance of asthma through the second decade of life is not caused by increased loss of established asthma in boys between 10 and 14 years of age. PMID- 11486788 TI - Application of fast field cycling NMR relaxometer to porous media. PMID- 11486789 TI - Some aspects of the fluid flow in a porous media in the microgravity conditions for the space plant production systems. PMID- 11486790 TI - Influence of orlistat on bone turnover and body composition. AB - OBJECTIVE: To investigate the influence of the pancreas lipase inhibitor orlistat (OLS) on calcium metabolism, bone turnover, bone mass, bone density and body composition when given for obesity as adjuvant to an energy- and fat-restricted diet. DESIGN: Randomized controlled double-blinded trial of treatment with OLS 120 mg three times daily or placebo for 1 y. SUBJECTS: Thirty obese subjects with a mean body mass index (BMI) of 36.9+/-3.7 kg/m(2) and a mean age of 41+/-11 y. Sixteen patients were assigned to OLS and 14 to placebo. MEASUREMENTS: Dual energy X-ray absorptiometry (DXA) measurements of bone mineral and body composition included total bone mineral content (TBMC), total bone mineral density (TBMD), lumbar spine BMC and BMD, forearm BMC and BMD, fat mass (FM), fat free-mass (FFM), percentage fat mass (FM%) as well as a DXA estimate of the body weight. Body composition (FM, FFM and FM%) was estimated by total body potassium (TBK). Indices of calcium metabolism and bone turnover included serum values of ionized calcium (Ca(++)), iPTH (parathyroid hormone), alkaline phosphatase, 25(OH)-vitamin D, 1,25(OH)(2) vitamin D and osteocalcin as well as fasting urinary ratios of hydroxyproline/creatinine and Ca/creatinine (fU-OHpr/creat, fUCa/creat). RESULTS: There were no significant differences between OLS and placebo groups as to any of the body composition variables (FFM, FM, FM%) at baseline or after 1 y treatment. Weight loss was of 11.2+/-7.5 kg in the OLS group and 8.1+/-7.5 kg in the placebo group (NS). The changes in FM and FM% were significant in both groups determined by DXA as well as by TBK, but the group differences between these changes were not significant. The composition of the weight loss was approximately 80% fat in both groups. FFM only changed significantly by DXA in the OLS group (-1.3 kg), but the difference from the placebo group was not significant. Forearm BMD in both groups, forearm BMC in the OLS group and TBMD in the placebo group fell discretely but significantly, but there were no significant group differences between the OLS and the placebo treated group. All biochemical variables except s-osteocalcin changed significantly after 1 y in the OLS group, disclosing a pattern of an incipient negative vitamin D balance, a secondary increase in PTH-secretion, and an increase in bone turnover with the emphasis on an increase in resorption parameters (fU-OHpr/creat, fUCa/creat). In the placebo group, only s 25(OH)vitamin D and fU-OHpr/creat changed significantly, but the pattern was also that of a deteriorated vitamin D status and an increase in PTH levels and bone turnover. The only biochemical variable which was significantly different between OLS and placebo groups after one year was the fU-OHpr/creat ratio, which increased from 12.0 to 20.1 in the OLS group but only from 10.9 to 1 3.2 in the placebo group. CONCLUSION: One year's treatment with OLS induces a lipid malabsorption which enhances a dietary weight loss without any significant deleterious effects on body composition. OLS induces a relative increase in bone turnover in favour of resorption, possibly due to malabsorption of vitamin D and/or calcium. However, no changes in bone mass or density are seen after 1 y of OLS treatment apart from those explained by the weight loss itself. Thus 1 y of OLS treatment seems safe from a 'bone preserving' point of view. A vitamin D and calcium supplement should be taken during the treatment. PMID- 11486791 TI - Surgery for orbital lesions. PMID- 11486792 TI - [Fear of dying after intra-keloid injection of triamcinolone acetonide and lidocaine: Hoigne's syndrome]. PMID- 11486793 TI - [Vulvodynia: a problem of definition]. PMID- 11486794 TI - [Formaldehyde related textile allergy in atopic patient]. PMID- 11486795 TI - A gene required for the specification of dorsal-ventral pattern in Drosophila appears to encode a serine protease. AB - The maternal effect gene snake is required for the establishment of the dorsal ventral axis during the embryonic development of Drosophila. The molecular cloning of the gene and analysis of a complementary DNA sequence suggest that the gene encodes a serine protease which is structurally similar to proteases involved in blood clotting, peptide processing, and complement fixation pathways. PMID- 11486796 TI - Reducing the progression of myopia with atropine: a long term cohort study of Olmsted County students. AB - BACKGROUND: Myopia is an important public health problem because it is common and is associated with increased risk for chorioretinal degeneration, retinal detachment, and other vision- threatening abnormalities. In animals, ocular elongation and myopia progression can be lessened with atropine treatment. This study provides information about progression of myopia and atropine therapy for myopia in humans. METHODS: A total of 214 residents of Olmsted County, Minnesota (118 girls and 96 boys, median age, 11 years; range 6 to 15 years) received atropine for myopia from 1967 through 1974. Control subjects were matched by age, sex, refractive error, and date of baseline examination to 194 of those receiving atropine. Duration of treatment with atropine ranged from 18 weeks to 11.5 years (median 3.5 years). RESULTS: Median followup from initial to last refraction in the atropine group (11.7 years) was similar to that in the control group (12.4 years). Photophobia and blurred vision were frequently reported, but no serious adverse effects were associated with atropine therapy. Mean myopia progression during atropine treatment adjusted for age and refractive error (0.05 diopters per year) was significantly less than that among control subjects (0.36 diopters per year)(P<.001). Final refractions standardized to the age of 20 years showed a greater mean level of myopia in the control group (3.78 diopters) than in the atropine group (2.79 diopters) (P<.001). CONCLUSIONS: The data support the view that atropine therapy is associated with decreased progression of myopia and that beneficial effects remain after treatment has been discontinued. PMID- 11486797 TI - Comparison of transcranial color Doppler imaging (TCDI) and transcranial Doppler (TCD) in children with sickle-cell anemia. AB - BACKGROUND: Transcranial Doppler (TCD) has been demonstrated to identify those at highest risk of stroke among children with sickle-cell disease. Based on a randomized clinical trial [Stroke Prevention in Sickle-Cell Anemia Trial (STOP)], which ended in 1997, the National Heart Lung and Blood Division of NIH has recommended TCD screening and chronic blood transfusion based on Nicolet TC 2000 dedicated Doppler (TCD). Studies performed using TCD imaging modalities need to be correlated to that used in the clinical trial to provide information for treatment decisions when screening with TCDI. OBJECTIVE: To correlate transcranial arterial time-averaged mean velocities obtained from an Acuson Transcranial Doppler Imaging to those obtained using the TCD as the gold standard for treatment decisions based on STOP. MATERIALS AND METHODS: A total of 29 children with sickle-cell disease, age 3-16 years, were studied at one of two scanning sessions using both techniques and a scanning protocol based on that used in STOP performed and read independently. The average difference in the measured velocities for each arterial segment was tested to determine difference from zero. Differences were compared before and after modifications to the TCDI technique were made to mimic the STOP protocol more closely. RESULTS: TCDI velocities were generally lower than TCD velocities for the same segment, but the difference was reduced (from 15 % to 10% for the middle cerebral artery) by modifications to the TCDI protocol. CONCLUSIONS: Measurements using the Acuson system are modestly lower than those obtained with dedicated Doppler using the Nicolet TCD. PMID- 11486798 TI - Infantile null cell acute lymphoblastic leukaemia (ALL) presenting with cerebellar tonsillar herniation. AB - We report a child with ALL who presented with acute neurological collapse and coning of the cerebellar tonsils as a result of massive brain infiltration by leukaemic cells. Such severe involvement of cerebral parenchyma by leukaemic cells is rare and cerebellar tonsillar herniation has not been reported. CNS parenchymal involvement with leukaemia is demonstrated using MRI and, for the first time, cranial US. PMID- 11486799 TI - Thymic ultrasound. I. Intrathymic anatomy in infants. AB - BACKGROUND: The gross appearance and echogenicity of the normal thymus have been described, but specific intrathymic anatomy has not been evaluated with imaging. SUBJECTS AND METHODS: The thymus from a 34-week-gestation stillborn male was examined with ultrasound, and the images were correlated with microscopic findings. Thymic ultrasound was performed in 12 infants without any evidence of thymic abnormality, severe infection, or chronic illness. Images were evaluated focusing on intrathymic parenchymal anatomy and compared to the liver, spleen, and thyroid gland. RESULTS: In the specimen thymus, ultrasound demonstrated connective-tissue septa as echogenic linear structures. The cortex was relatively hypoechoic, whereas the medulla was echogenic. The blood vessels within the septa were seen as discrete echoes. In vivo ultrasound demonstrated multiple branching echogenic linear structures and foci throughout the parenchyma, representing connective-tissue septa or blood vessels within the septa. The normal thymus was easily differentiated from the liver, spleen, and thyroid glands. CONCLUSION: Ultrasound is capable of demonstrating intrathymic anatomy, including the medulla, cortex, septa, and blood vessels in a fresh specimen. In vivo connective tissue septa and blood vessels in the thymic parenchyma produce a unique echo pattern. PMID- 11486800 TI - Thymic ultrasound. II. Diagnosis of aberrant cervical thymus. AB - BACKGROUND: The aberrant cervical thymus is an uncommon entity to be considered in the differential diagnosis of neck masses in children, so a preoperative diagnosis has rarely been made. OBJECTIVE: The purpose of this study was to assess the ultrasound features of aberrant cervical thymus and determine if recognition of the previously described unique intrathymic echo pattern allows us to make a specific diagnosis. MATERIALS AND METHODS: We present four patients with an aberrant cervical thymus and one patient with a cervical thymic cyst. All five patients had ultrasound, which was evaluated with special attention to the detailed echo pattern of the mass. In two patients, the mass was surgically removed. Four patients had MRI, which was evaluated. Medical records were reviewed. RESULTS: In all cases, the mass was painless and nontender. All occurred in the expected path of the thymopharyngeal duct. In four patients, ultrasound demonstrated multiple echogenic linear structures and foci previously described as characteristic echo pattern of normal thymic tissue. In one patient, ultrasound showed a large cystic mass and echogenic solid component superiorly with a characteristic echo pattern of normal thymus. CONCLUSIONS: The ultrasound appearance of thymic tissue is unique, allowing a specific diagnosis of aberrant cervical thymus. Biopsy or other imaging is probably not indicated when it presents with typical clinical features. PMID- 11486801 TI - Mycotic aneurysm of the thoracic aorta presenting as pneumonia. AB - Mycotic aneurysms of the thoracic aorta rarely occur in children. We report an unusual case of a mycotic aneurysm of the descending aorta in a 4-year-old boy presenting with respiratory tract infection, which was rapidly complicated by atelectasis of the left lung. The patient's mycotic aortic aneurysm was diagnosed by contrast-enhanced spiral CT, whereas conventional chest radiographs did not detect its presence. An unsuspected mild aortic coarctation was also diagnosed at the time of admission. This case demonstrates that an aortic aneurysm may clinically and radiologically manifest itself with respiratory tract infection and atelectasis and that contrast-enhanced spiral CT is a fast and powerful tool for establishing the diagnosis. PMID- 11486802 TI - Ahaustral colonic segment following trauma: a possible sign of seromuscular tear. AB - Although traumatic seromuscular tears of the colon are often managed conservatively, it is pertinent to detect the tear since it poses a risk for acute or delayed perforation. Unfortunately, clinical findings are inadequate for detecting and monitoring this lesion. Therefore, this complication has not been recognized prior to laporatomy. We have noted that an ahaustral colonic segment following trauma may be a potential marker for seromuscular tear. PMID- 11486803 TI - Colo-colic intussusception in an infant with pneumatosis. AB - The finding of a colo-colic intussusception with pneumatosis intestinalis is described with imaging. This was confirmed at surgery together with pathological confirmation of pneumatosis intestinalis. This rare association occurred in an infant with Peters' anomaly who was taking oral steroids. PMID- 11486804 TI - The screening pelvic radiograph in pediatric trauma. AB - BACKGROUND: Pelvic radiographs are routinely obtained in adult trauma to optimise early management. In adults, pelvic fractures are associated with high early transfusion requirement, high injury severity scores and an increased incidence of other abdominal and thoracic injuries. It is unclear whether this holds true in children. OBJECTIVE: To determine whether the screening pelvic radiograph is necessary in paediatric trauma. MATERIALS AND METHODS: The notes of all patients who presented after trauma to the Starship Children's Hospital and were triaged to the resuscitation room during 1997 were reviewed. Results of initial radiography were obtained and correlated with later imaging. RESULTS: Our review of 444 injured children seen over a period of 1 year revealed that of 347 children who had screening pelvic radiographs, only 1 had a pelvic fracture. The fracture in this child was clinically apparent and required no specific treatment. CONCLUSIONS: The presence of a pelvic fracture is rare in injured children. By omitting screening pelvic radiographs there are potential benefits, including reduced radiation exposure to children and cost savings. Uninterpretable or abnormal clinical examination or haematuria requires further investigation, but routine screening for pelvic fracture is unnecessary. PMID- 11486805 TI - Imaging features of pancreatoblastoma. AB - BACKGROUND: Pancreatoblastoma is a rare tumour of childhood. Reports of the imaging appearances are limited. OBJECTIVE: To define the imaging features of pancreatoblastoma by analysis of four previously unreported cases and review of the literature. MATERIALS AND METHODS: Findings at CT (n = 4), US (n = 3) and MRI (n = 2) were retrospectively reviewed in four patients with pancreatoblastoma. A Medline search was performed to identify relevant literature. RESULTS: Pancreatoblastoma arises most frequently in the body and/or tail, or involves the entire pancreas. Ultrasonography, CT and MRI show variable imaging features, but should in most cases permit preoperative distinction of pancreatoblastoma from other tumours that occur in this region in infancy and childhood. Detection of metastases in the liver, lymph nodes and peritoneal cavity is not significantly better with any one of these three modalities. CONCLUSION: Preoperative imaging with US, CT and/or MRI will usually suggest a correct diagnosis of pancreatoblastoma. Contrary to previous reports, the tumour arises in the pancreatic head in a minority of cases. PMID- 11486806 TI - Diffuse venous malformations of the upper limb: morphologic characterization by MRI and venography. AB - OBJECTIVES: To define the morphologic abnormalities in patients presenting with diffuse pure venous malformations (VM) of the upper extremity. SUBJECTS AND METHODS: A retrospective review of MRI and venography was performed on five patients, aged 6 months to 20 years, with extensive VM of the upper limbs. Abnormalities of major conducting veins were categorized as varicosities, stenoses, and asymmetrical pouches; anomalous venous spaces were classified into confluent lakes, interconnecting channels and spongelike plexiform networks. MRI and venographic data were reviewed separately and then simultaneously in order to establish correlation between types, location, and extent of lesions. RESULTS: In all patients, the percentage of replacement of normal tissues by VM was shown by MRI to be significantly higher in the distal limb than in the proximal limb. Involvement of multiple tissue layers was seen in all cases, including, with a decreasing rate, muscles, tendons, interosseous membrane of the forearm, and bone. Venography showed superficial varicosities, frequently associated with stenoses and assymetric pouches in all patients. Interconnecting channels and venous lakes were noted in half of the segments, typically in muscle and other deep locations, and subcutaneous spongelike lesions were seen in two patients. MRI provided a more accurate evaluation of tissue extent. Venograms better demonstrated morphological details and provided more information about the venous drainage. Direct comparison of MR images with venograms helped to identify and characterize venous lesions on cross-sectional MR data. CONCLUSION: Diffuse VM of the upper extremity are most extensive distally, and all tissues layers can be involved, each with a characteristic morphologic appearance. The morphology of different components of the VM is related to the nature of the surrounding tissue. PMID- 11486807 TI - Percutaneous transfemoral testicular vein embolisation in the treatment of childhood varicocoele. AB - BACKGROUND: Controversy surrounds the early treatment of childhood varicocoele and its role in the prevention of testicular atrophy and male infertility. Various techniques exist, all with varying degrees of success. OBJECTIVES: To show that percutaneous transfemoral testicular vein embolisation is an effective alternative when compared to the conventional open surgical approach. MATERIALS AND METHODS: A retrospective review examining 48 boys (aged 9-18 years; mean 13.2 years) who were treated with transcatheter testicular vein embolisation between 1985 and 1999. Follow-up took the form of out-patient clinical assessment and a telephone questionnaire. Patients were graded as 'good', 'moderate' or 'poor', according to various criteria. RESULTS: Of the 48 patients, 43 (90%) had satisfactory embolisations. Thirty-eight (88 %) had a 'good' clinical outcome at follow-up. There were five technical failures due to a combination of abnormal venous anatomy and severe venospasm. CONCLUSIONS: We believe that where the expertise necessary for testicular embolisation is available, it should be offered as the intervention of first choice. Surgery should be reserved for the rare cases where embolisation is not possible or when recurrence has occurred. PMID- 11486808 TI - Tumor volume or dynamic contrast-enhanced MRI for prediction of clinical outcome of Ewing sarcoma family of tumors. AB - BACKGROUND: The identification of risk factors that predict poor clinical outcome at the time of diagnosis could lead to intensified early therapy and improved outcome for pediatric patients with Ewing sarcoma family of tumors (ESFT). OBJECTIVE: To compare the effectiveness of static magnetic resonance (MR) imaging measurements of tumor volume with variables obtained by dynamic contrast-enhanced MR imaging (DEMRI) in predicting ESFT outcome. METHODS: MR examinations that included DEMRI were retrospectively reviewed. The analyses included 45 examinations of 21 patients with ESFT (performed from 1992 to 1996). Tumor volumes were measured on the static MR images, and the regions of interest were selected for DEMRI analysis. The relationships of static MR imaging and DEMRI variables with the probability of progression-free survival (PFS) and disease free survival (DFS) were determined. RESULTS: Larger tumor volume at the time of diagnosis predicted poorer PFS and DFS estimates. No DEMRI variable predicted outcome. CONCLUSION: Determination of tumor volume by static MR imaging at the time of diagnosis is a simple and reliable method of predicting the clinical outcome of patients with ESFT. DEMRI is not as reliable a technique as static MR imaging for predicting the outcome of these patients. PMID- 11486809 TI - Juvenile rheumatoid arthritis of the knee: evaluation with contrast-enhanced color Doppler ultrasound. AB - BACKGROUND: Contrast-enhanced color Doppler ultrasonography is a non-radiation bearing tool that can be of value for assessment of inflammatory and vascular synovial changes in juvenile rheumatoid arthritis (JRA). OBJECTIVES: To evaluate the effect of contrast-enhanced color Doppler ultrasound (US) in the evaluation of synovial changes in the knees of children with JRA. MATERIALS AND METHODS: Sagittal color Doppler sonograms of 31 knees in 22 patients with JRA and of 10 knees in 5 control subjects were obtained before (at baseline) and after (at peak contrast phase) intravenous injection of SHU 508. Images were assessed for overall mean pixel intensity within the synovial tissue and for peak enhancement ratios [[(mean pixel intensity values at maximum contrast enhancement-unenhanced mean pixel intensity values)/unenhanced mean pixel intensity values] x 100]. The joints were classified into three groups by clinical/laboratory criteria: group A (active disease in the knee), n = 9; group B (quiescent disease with serum chemistry levels of active disease), n = 12 and group C (remission disease), n = 10. RESULTS: Mean color pixel intensity values were markedly increased by the use of US contrast agents in groups A (P = 0.004) and B (P = 0.0001), did not reach statistical significance in group C (P = 0.06) and remained essentially unchanged in the control group (P = 0.25). Enhancement ratios for the three groups of JRA patients were not different (P = 0.38) (mean +/- SD, 720% +/- 402 for group A, 731% +/- 703 for group B and 314% +/- 263 for group C). CONCLUSION: Contrast enhanced color Doppler imaging holds promise for the detection of active synovial inflammatory disease in subclinical cases of JRA, thereby allowing earlier treatment and improved clinical outcome. PMID- 11486810 TI - An influence of stress protein CSP 310 and antiserum against this protein on lipid peroxidation in cereal mitochondria. AB - It is determined that an addition of an anti-CSP 310 antiserum to isolated winter wheat and maize mitochondria caused more significant increasing of spontaneous lipid peroxidation than the addition of stress protein CSP 310. It is shown that, at function of different mitochondrial respiratory chain complexes, the lipid peroxidation in winter wheat and maize mitochondria take place with different intensities. Under the functioning of mitochondrial respiratory chain complex IV, the maximum output of lipid peroxidation products, dienic conjugates is detected. The presence of antiserum against CSP 310 in incubation media induces lipid peroxidation more than the presence of CSP 310 in mitochondria isolated from stressed plants under these conditions. Based on data obtained, it is possible to conclude that in vivo endogenous CSP 310, during a cold stress, has an antioxidant activity the same as other known uncoupling proteins. PMID- 11486811 TI - Immunochemical control of the species origin of intestinal mucosa used for heparin purification. AB - Species specific antisera against bovine, ovine, and porcine serum albumin were produced in order to control the absence of bovine, ovine, or caprine tissues in the porcine intestinal mucosa used for heparin production. Two immunoassays were developed. An enzyme linked immunosorbent assay (ELISA) was very sensitive down to 1 ng/mL bovine albumin or 10 ppm bovine intestinal mucosa in porcine intestinal mucosa. For routine control, a more convenient single radial immunodiffusion assay (SRID) was found suitable to detect 2 microg/mL albumin or 3 p 1000 bovine, ovine, or caprine intestinal mucosa in porcine intestinal mucosa. Conditions of extraction of albumin from intestinal mucosa were optimized and a CV % of 4.1 was obtained for its quantitation. Due to higher albumin concentrations, detection of bovine hashed gut and lung was more sensitive (1.5 and 0.9 p 1000, respectively). Using antisera raised against porcine albumin the SRID can be applied to certify the porcine origin of the intestinal mucosa used for heparin purification and to control its adequate conservation before analysis. PMID- 11486812 TI - Validity and use of a non-parallel insulin assay for pharmacokinetic studies of the rapid-acting insulin analogue, insulin aspart. AB - A radioimmunoassay (RIA) for insulin was validated for reliable measurement of the human insulin analogue, insulin aspart, by correction of non-linear measurements. Specificity was equivalent for several species of insulin, except insulin aspart. A non-linear hyperbolic model fitted insulin aspart with a correction formula for non-linearity of: z = 1,503y/ (1,398 - y), where y denotes measured concentration and z denotes true concentration. Matrix-effects were insignificant for human, porcine, and canine heparin-plasma and for human and porcine serum. The coefficient of variation was below 15% for 80-800 pmol/L human and porcine insulin and for 80-600 pmol/L insulin aspart. The limit of detection for insulin aspart was 11.5 pmol/L with a lower limit of quantification of 17.5 pmol/ L. Dilution of serum with Pharmacia dilution media introduced no significant error. In conclusion, this paper demonstrates that a non-parallel radioimmunoassay can be used to estimate accurate concentrations of insulin aspart. PMID- 11486813 TI - Quantitation of Leishmania infantum in tissues of infected BALB/c mouse by sandwich ELISA. AB - In this report, a sandwhich ELISA was developed to quantify spleen and liver burdens from L. infantum-infected BALB/c mice. Amastigote antigens obtained following Nonidet P40 extraction of parasite-harbouring tissues were captured by anti-L. infantum human IgG insolubilized onto microtiter plate and subsequently revealed with anti-L. infantum F(ab)' fragments labelled with peroxidase. The method was easy to perform, precise and capable to specifically and accurately detect 5 x 10(4) amastigotes/100 mg tissue. Parasite burdens from infected BALB/c mice, in various conditions, were measured by ELISA and Giemsa-stained touch imprint reference methods, and compared. Both techniques agreed well with close values for liver burdens, but the spleen loads measured by the ELISA were, on average, 10.7 times higher than those calculated from imprints. This difference was attributed partly to the underestimation brought by Stauber's formula. However, it did not preclude the usefulness of this newly developed test, since results obtained in kinetics studies and evaluation of the efficiency of leishmanicidal drugs allowed us to draw identical conclusions. PMID- 11486814 TI - Comparison of some methods for determing cutoff values for serological assays: a retrospective study using the fluorescence polarization assay. AB - Different methods for determining cutoff values between positive and negative results for serological assays have been developed over time. Comparisons of some these methods show that five (Receiver Operating Characteristics, Frequency Distribution, and the mean, median and mode of the 100th percentile of a disease free group of data) resulted in similar sensitivity (99.11%) and specificity values (99.21% to 99.58%). However, the Receiver Operating Characteristic analysis was considered more suitable, due to the ease of data manipulation and computer analysis, providing flexibility for different field applications. PMID- 11486815 TI - A specific and ultrasensitive chemiluminescent sandwich ELISA test for the detection and quantitation of pneumolysin. AB - A chemiluminescent sandwich ELISA test has been developed for the detection and quantitation of pneumolysin. The test is based on a mouse monoclonal as the capture antibody and on rabbit polyclonal IgGs as detection antibodies, in combination with an anti-rabbit IgG alkaline phosphatase conjugate. The estimated detection limit of the purified recombinant toxin in phosphate-buffered saline with 0.05% Triton X-100 is around 5 pg ml(-1), with averaged intra- and inter assay variation coefficients of 7% and 13.5%, respectively. The assay has been applied to the quantitation of pneumolysin in pneumococcal isolates, providing, for the first time, a direct measurement of the amount of the toxin produced by different strains, a variation has been found in their pneumolysin content. The test is highly specific as no other purified toxins or human pneumonia- or meningitis-associated bacteria yielded false-positive results. This specific and highly sensitive method could help in the diagnosis of human infections. PMID- 11486816 TI - Enzyme immunoassay for conjugated 7alpha-hydroxy-3-oxo-4-cholenoic acid in human urine. AB - A microplate enzyme immunoassay (EIA) was developed for the measurement of glycine- and taurine-conjugated 7alpha-hydroxy-3-oxo-4-cholenoic acids (CDCA delta4-3-one) in human urine. The antiserum was prepared by immunizing rabbits with N-(7alpha-hydroxy-3-oxo-4-cholen-24-oyl)-3-aminopropionic acid--bovine serum albumin conjugate. A colorimetric EIA was established using horseradish peroxidase-labeled antigen having a shorter bridge length than that of the immunogen, and 3, 3', 5, 5'-tetramethylbenzidine /hydrogen peroxide for the measurement of the enzyme activity. The reactivities of the antiserum for glycine and taurine conjugates of CDCA-delta4-3-one was almost the same. The specificity of the antiserum was investigated by determining the cross-reactivities of various bile acids and related compounds. An appropriate dose-response curve for conjugated CDCA-delta4-3-one was obtained in the range of 0.05-10 ng/well. This method was used for direct analysis of conjugated CDCA-delta4-3-one in urine of healthy infants and patients with liver diseases. PMID- 11486817 TI - Development of an enzyme immunoassay for a stable amidated analog of the hemoregulatory peptide acetyl-Ser-Asp-Lys-Pro. AB - The tetrapeptide Acetyl-Ser-Asp-Lys-Pro (AcSDKP) has been shown to protect hematopoietic stem cells from the toxicity of anticancer chemotherapies. Since its pharmacological efficacy is limited by a rapid degradation by Angiotensin-I Converting Enzyme (ACE), AcSDKP analogs resistant to ACE have been synthesized. One of these compounds (AcSDKP-NH,) differs from the native AcSDKP by amidation of the C-terminus. Further evaluations of this molecule require an analytical method in order to characterize its pharmacokinetic profile. We report, here, the development of a highly specific and sensitive enzyme immunoassay (EIA) for AcSDKP-NH, thatdoes not cross-react with endogenous or exogenous AcSDKP. Using AcSDKP-NH2-acetylcholinesterase conjugate as a tracer, rabbit specific antiserum and microtiter plates coated with goat anti-rabbit immunoglobulins, this EIA allows the determination of AcSDKP-NH2 with limits of quantitation of 1 nM in mouse plasma and 100 pmol/g in tissues. Intra-day and inter-day coefficients of variations were less than 20%. The method was successfully applied to a pharmacokinetic study in order to compare plasma and tissue profiles of AcSDKP NH2 and AcSDKP. Plasma AcSDKP-NH2 levels were found higher than those of AcSDKP, with AUCinf and Cmax values, respectively, 26- and 10-fold higher than that of AcSDKP. PMID- 11486818 TI - Anti-HBc IgG subclasses in different populations by comparing a variety of ELISA plates. AB - The number of IgG subclasses for hepatitis B virus (HBV) core antigen (anti-HBc), demonstrated for HBV-infected individuals, was measured by enzyme-linked immunosorbent assay (ELISA). Four commercially available hepatitis B core antigen (HBcAg) plates and one prepared plate were tested for ELISA sensitivity by the detection of 14 serum samples drawn from HBV chronic carriers, cured patients, vaccinees, and non-infected individuals. Differences in optical density (OD) values were obtained by comparing data gathered from the five plate types, suggesting that different plates may have different binding capabilities for each anti-HBc IgG subclass and, thus, contribute to the different ELISA sensitivities. Of these plates, the GB plate showed the most obvious absorbance changes for anti HBc subclasses in different populations. These data also indicated different patterns for IgG-specific subclasses for various populations. For HBsAg+ carriers, the OD for IgG1 was greater than for IgG3. By contrast, the OD for IgG3 was higher than that for IgG1 in those subjects who were negative for HBsAg. PMID- 11486819 TI - Studies on the mechanism of multiple drug allergies. Structural basis of drug recognition. AB - The multiple drug allergy syndrome, that is, allergic recognition of a variety of drugs that may be both pharmacologically and structurally different, has been little studied and, consequently, the underlying mechanism(s) is unknown. The molecular basis of drug recognition by IgE antibodies found in the sera of subjects exhibiting multiple allergic drug sensitivities was studied by direct binding and quantitative hapten inhibition assays in experiments employing a wide range of carefully selected drugs and other chemicals. Drug recognition was shown to be related to the presence of tertiary and quaternary mono-, di- and trialkyl amino groups, but only if the alkyl groups were 'small' viz., methyl or, perhaps, ethyl. Primary, secondary, and tertiary (with R = 'large' alkyl) groups showed no direct antibody binding or antibody inhibitory activities. Near-neighbour effects of amide and hydroxyl groups appeared to promote weaker antigenic recognition. Results indicate that the antibody recognition and clinical drug allergy spectra of at least some subjects with multiple drug allergies are due to wide ranging immunological cross-reactivities with drugs containing tertiary amino and quaternary ammonium groups which are present in many different pharmacologically active agents. Separate populations of antibodies to other non-cross reacting drugs, for example, beta-lactam antibiotics, may also be present in the sera of such subjects. PMID- 11486820 TI - An influence of antiserum against winter wheat stress uncoupling protein, CSP 310, on energetic activity of some plant species mitochondria. AB - It is determined that addition of an anti-CSP 310 antiserum to isolated mitochondria of cereals (winter rye, winter wheat, and maize) caused an increasing of mitochondrial respiratory control. In a similar manner, addition of this antiserum to isolated pea mitochondria did not cause this effect. It is shown that coupling effect of antiserum is not dependent upon the presence of bovine serum albumin in mitochondria incubation media. Therefore, these results show that the mechanism of oxidation and phosphorylation uncoupling, and participation of immunochemically related CSP 310 proteins that can be precipitated by anti-CSP 310 antiserum is specific for cereals. PMID- 11486821 TI - High-performance liquid chromatographic assay to determine the plasma levels of HIV-protease inhibitors (amprenavir, indinavir, nelfinavir, ritonavir and saquinavir) and the non-nucleoside reverse transcriptase inhibitor (nevirapine) after liquid-liquid extraction. AB - A single HPLC assay was developed for therapeutic drug monitoring of 5 HIV protease inhibitors (indinavir, amprenavir, saquinavir, ritonavir, nelfinavir) and a non-nucleoside reverse transcriptase inhibitor (nevirapine) in human plasma. After liquid-liquid extraction in a mixture ethyl acetate-hexane, compounds are separated on a C18 column with a gradient elution of solvent A [acetonitrile and 0.025 M tetramethylammonium perchlorate in 0.2% aqueous trifluoroacetic acid (55:45 (v/v))] and solvent B [methanol and 0.025 M tetramethylammonium perchlorate in 0.2% aqueous trifluoroacetic acid (55:45 (v/v))]. The compounds are detected at various wavelengths: 320 nm (nevirapine), 259 nm (indinavir), 254 nm (amprenavir, nelfinavir, saquinavir) and 239 nm (ritonavir). The intra-day and inter-day precision and accuracy are lower than 15%. The limits of quantitation are 0.05 mg/l (amprenavir), 0.2 mg/l (indinavir, saquinavir, nelfinavir) and 0.4 mg/l (ritonavir, nevirapine). This method which allows to estimate simultaneously plasma levels of protease inhibitors and nevirapine can be used for therapeutic drug monitoring. PMID- 11486822 TI - Automated solid-phase extraction method for the determination of atovaquone in capillary blood applied onto sampling paper by rapid high-performance liquid chromatography. AB - A bioanalytical method for the determination of atovaquone in 100 microl blood spots by solid-phase extraction and high-performance liquid chromatography has been developed and validated. Atovaquone was extracted from the sampling paper in 0.2 M phosphoric acid and a structurally similar internal standard was added with acetonitrile before being loaded onto a C8 end-capped solid-phase extraction column. Atovaquone and internal standard were analysed by high-performance liquid chromatography on a C18 J'Sphere ODS-M80 (150 x 4.0 mm) column with mobile phase acetonitrile-phosphate buffer, 0.01 M, pH 7.0 (65:35, v/v) and UV detection at 277 nm. The intra-assay precision was 2.7% at 12.00 microM and 13.5% at 1.00 microM. The inter-assay precision was 3.3% at 12.00 microM and 15.6% at 1.00 microM. The lower limit of quantification was 1.00 microM. The limit of detection was 0.50 microM. PMID- 11486823 TI - Gas chromatographic determination of catecholestrogens following isolation by solid-phase extraction. AB - A sensitive and specific assay for the determination of the catecholestrogens 2 hydroxyestradiol (2-OHE2) and 4-hydroxyestradiol (4-OHE2) using gas chromatography with electron-capture detection (GC-ECD) is described. The formation of 2- and 4-OHE2 was assessed following activation of 17beta-estradiol in the microsomal fraction of female rat livers. The analytes were isolated by solid-phase extraction, derivatized to their heptafluorobutyryl esters with heptafluorobutyric acid anhydride, and subjected to solvent exchange prior to analysis; this resulted in minimal chromatographic interference, long column life, and stable derivatized analytes. Derivatized catechols were separated and confirmed with dual column chromatography (DB-5 and DB-608) and quantitated using GC-ECD. The DB-608 column was preferred for quantitation as it provided better 4 OHE2 resolution from interference. Key validation parameters for the assay include sensitivity, intra- and inter-assay precision, and accuracy. Instrument sensitivity and limits of detection (LOD) and quantitation (LOQ) were determined statistically from fortification data approaching expected limits. For 2-OHE2 and 4-OHE2, respective values for these parameters were; instrument sensitivities of 0.4 and 0.7 pg, LODs of 0.8 and 1.3 ng/mg, and LOQs of 2.6 and 4.3 ng/mg. PMID- 11486824 TI - Enantiomeric separation of metoprolol and alpha-hydroxymetoprolol by liquid chromatography and fluorescence detection using a chiral stationary phase. AB - A sensitive and stereoselective high-performance liquid chromatographic assay for the determination of the enantiomers of metoprolol (R- and S-) and the diastereoisomers of alpha-hydroxymetoprolol (IIA, IIB) in plasma is reported. Chromatography involved direct separation of enantiomers using a Chirobiotic T bonded phase column (250 x 4.6 mm) and a mobile phase consisting of acetonitrile methanol-methylene chloride-glacial acetic acid-triethylamine (56:30:14:2:2, v/v). Solid-phase extraction using silica bonded with ethyl group (C2) was used to extract the compounds of interest from plasma and atenolol was used as the internal standard. The column effluent was monitored using fluorescence detection with excitation and emission wavelengths of 225 and 310 nm, respectively. S Metoprolol, R-metoprolol, IIB and IIA eluted at about 5.9, 6.7, 7.3 and 8.2 min without any interfering peaks. The calibration curve was linear over the range of 0.5 to 100 ng/ml for each isomer of metoprolol and 1 to 100 ng/ml for each isomer of alpha-hydroxymetoprolol (IIA & IIB). The mean intra-run accuracies were in the range of 96.2 to 114% for R-metoprolol, 94.0 to 111% for S-metoprolol, 90.2 to 110% for IIA, and 94.6 to 106% for IIB. The mean intra-run precisions were all in the range of 2.2 to 12.0% for R-metoprolol, 2.1 to 11.1% for S-metoprolol, 1.9 to 14.5% for IIA, and 3.2 to 11.0% for IIB. The lowest level of quantitation for the enantiomers of metoprolol was 0.5 ng/ml and 1.0 ng/ml for alpha-hydroxymetoprolol (IIA and IIB). The absolute recoveries for each analyte was > or = 95%. The validated method accurately quantitated the enantiomers of parent drug and metabolite after a single dose of an extended release metoprolol formulation. PMID- 11486825 TI - Protein adsorption on histidyl-aminohexyl-Sepharose 4B. I. Study of the mechanistic aspects of adsorption for the separation of human serum albumin from its non-enzymatic glycated isoforms (advanced glycosylated end products). AB - The characteristics of albumin adsorption on histidyl-aminohexyl-Sepharose 4B were investigated. In particular, the adsorption capacity of the gel was studied as a function of conductivity and pH of the running buffer. The adsorption was maximum at low salt concentration around neutral pH, involving electrostatic and hydrophobic interactions. Kinetic aspects were also investigated. Dissociation constant (KD) and maximum capacity (Qx) were, respectively, estimated to be 4.5 x 10(-5) M (medium affinity) and 93.3 mg (high capacity) of human serum albumin per ml of adsorbent. According to these preliminary results, separation of HSA and its non-enzymatically glycated isoforms (conventionally named advanced glycated end products: AGEs) was achieved. Chromatographic potential of this separation tool is discussed. PMID- 11486826 TI - Protein adsorption on histidyl-aminohexyl-Sepharose 4B. II. Application to the negative one-step affinity purification of human beta2-microglobulin and immunoglobulin G. AB - The adsorption of two human proteins, beta2-microglobulin and Immunoglobulin G, from uremic patient's blood ultrafiltrate and plasma, respectively, was investigated on the histidyl-aminohexyl-Sepharose 4B adsorbent. Both target proteins could be adsorbed on the gel through a low affinity for immobilized histidine ligand. However, a fine adjustment of the operating conditions (ionic strength, buffer, pH) prevented their adsorption and thus allowed their "negative affinity" purification (purity estimated by silver nitrate SDS-PAGE) by the removal of the contaminating proteins. This simple and efficient method provides purification under gentle chromatographic conditions and a further characterization of both molecules. PMID- 11486827 TI - Rapid and simple high-performance liquid chromatographic determination of nimesulide in human plasma. AB - A high-performance liquid chromatographic method for the quantitation of nimesulide in human plasma is presented. The method is based on protein precipitation with methanol and reversed-phase chromatography with spectrophotometric detection at 404 nm. The separation was performed on a Nucleosil 120-5 C18, 50 x 4-mm I.D. column and the mobile phase consisted of acetonitrile-methanol-15 mM potassium dihydrogenphosphate buffer, pH 7.3 (30:5:65, v/v). Only 250 microl of plasma are used for sample preparation and no internal standard is necessary. The limit of quantitation is 80 ng/ml and the calibration curve is linear up to 10,000 ng/ml. More than 20 samples can be analysed within 1 h. Within-day and between-day precision expressed by relative standard deviation is less than 5% and inaccuracy does not exceed 8%. The assay was used for pharmacokinetic studies. PMID- 11486828 TI - Simplified in-line sample preparation for amino acid analysis in carbohydrate containing samples. AB - This report describes a new, automated chromatographic procedure eliminating carbohydrates from amino acid samples prior to their analysis by anion-exchange chromatography and integrated amperometric detection. In the first step, a sample is brought onto a short cation-exchange column (trap column) in hydrogen form. Carbohydrates are passing through this column, while only amino acids are retained. Subsequently, the cation-exchange column, holding the amino acid fraction, is switched in-line with the gradient pump and separator column. The mobile phase used at the beginning of the separation (NaOH; pH 12.7) transfers amino acids from the trap column onto the anion-exchange column and the amino acid separation is completed without any interference by carbohydrates. All common amino acids are recovered following the carbohydrate removal step. The average value of their recovery is 88.1%. The calibration plots were tested between 12.5 and 500 pmol (amounts injected). The mean value of correlation coefficients of calibration plots was calculated as 0.99. The mean value of relative standard deviations from five replicates was 3.9%. The usefulness of the method is illustrated with two chromatograms of a carrot juice sample obtained before and after the in-line removal of carbohydrates. PMID- 11486829 TI - Lytic enzyme complex of an antagonistic Bacillus sp. X-b: isolation and purification of components. AB - Bacillus sp. X-b, a biocontrol agent against certain plant pathogenic fungi, secretes a complex of hydrolytic enzymes, composed of chitinase, chitosanase, laminarinase, lipase and protease. Homogenized mycelium of basidiomycete Macrolepiota procera induced activities of these enzymes more effectively than colloidal chitin or partially purified cell walls of another basidiomycete Polyporus squamosus. Subjected to a multi-step purification, the specific activity of chitinase increased 36-fold, chitosanase 69-fold, lipase 44-fold and laminarinase 15-fold. Partially purified chitinase showed two major bands with molecular masses of 46,000 and 35,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis while chitosanase and lipase appeared as single bands with molecular masses of 27,000 and 62,000, respectively. PMID- 11486830 TI - Pre-column derivatization high-performance liquid chromatographic method for determination of cysteine, cysteinyl-glycine, homocysteine and glutathione in plasma and cell extracts. AB - A sensitive high-performance liquid chromatographic method for quantification of sulphydryl and disulfide amino acids in human plasma using ultra violet spectrophotometric detection was developed. Precolumn derivatization with 5,5' dithio-bis-nitrobenzoic acid (DTNB) and an optional pre-derivatization reaction with dithiothreitol allowed both quantitative reduction of disulfides for measurement of total amino acid levels and the measurement of the reduced forms. A dynamic range of 500 nmol/l-750 micromol/l allowed the major analytes of interest to be quantified in plasma without sample dilution. The assay is a sensitive and precise method for the determination of sulphydryl and disulfide amino acids in plasma and cell extracts. PMID- 11486831 TI - Simultaneous analysis of the di(2-ethylhexyl)phthalate metabolites 2 ethylhexanoic acid, 2-ethyl-3-hydroxyhexanoic acid and 2-ethyl-3-oxohexanoic acid in urine by gas chromatography-mass spectrometry. AB - A gas chromatographic-mass spectrometric method was developed for the quantitative analysis of the three Di(2-ethylhexyl)phthalate (DEHP) metabolites, 2-ethylhexanoic acid, 2-ethyl-3-hydroxyhexanoic acid and 2-ethyl-3-oxohexanoic acid in urine. After oximation with O-(2,3,4,5,6-pentafluorobenzyl)-hydroxylamine hydrochloride and sample clean-up with Chromosorb P filled glass tubes, all three organic acids were converted to their tert.-butyldimethylsilyl derivatives. Quantitation was done with trans-cinnamic acid as internal standard and GC-MS analysis in the selected ion monitoring mode (SIM). Calibration curves for all three acids in the range from 20 to 1,000 microg/l showed correlation coefficients from 0.9972 to 0.9986. The relative standard deviation (RSD) values determined in the observed concentration range were between 1.3 and 8.9% for all three acids. Here we report for the first time the identification of 2-ethyl-3 hydroxyhexanoic acid and 2-ethyl-3-oxohexanoic acid in human urine next to the known DEHP metabolite 2-ethylhexanoic acid. In 28 urine samples from healthy persons we found all three acids with mean concentrations of 56.1 +/- 13.5 microg/l for 2-ethylhexanoic acid, 104.8 +/- 80.6 microg/l for 2-ethyl-3 hydroxyhexanoic acid and 482.2 +/- 389.5 microg/l for 2-ethyl-3-oxohexanoic acid. PMID- 11486832 TI - Quantification of daunorubicin and daunorubicinol in plasma by capillary electrophoresis. AB - Capillary electrophoresis (CE) with laser-induced fluorescence detection was applied to quantify daunorubicin and daunorubicinol in plasma. Separation was carried out in a 47 cm x 50 microm I.D. fused-silica capillary, with a running buffer. pH 5 containing 60 microM spermine and 70% acetonitrile. Sample preparation was done either by protein precipitation with acetonitrile or by liquid-liquid extraction. The assay can be applied in a concentration range from 40 mg/l down to 2 microg/l for daunorubicin and from 1 mg/l to 2 microg/l for daunorubicinol. Precision and accuracy were between 2.9 and 14.5% (n=6) on 1 day and between 1.0 and 14.7% from day to day (n=6) for both analytes. Thus, the CE method enables precise and accurate quantification of daunorubicin and daunorubicinol in small sample volumes over a wide concentration range. PMID- 11486833 TI - Improvement of ethyl glucuronide determination in human urine and serum samples by solid-phase extraction. AB - An improved method for the determination of ethyl glucuronide (EtG) in human serum and urine was developed using solid-phase extraction (SPE) and gas chromatography (GC) with mass spectrometric detection (MS). EtG was isolated from serum and urine using aminopropyl SPE columns after deproteination with perchloric acid and hydrochloric acid, respectively. The chromatographic separation was performed on a DB 1701 fused-silica column. At a signal-to-noise ratio of 3:1, a quantification limit of 173 and 560 ng/ml and a detection limit of 37 and 168 ng/ml could be determined for serum and urine, respectively. This indicates high specificity and sensitivity of the described method. The mean absolute recovery was approximately 85%, while intra- and inter-day precision of the assay were all less than 7.5%. The linearity of the calibration curves was satisfying as indicated by correlation coefficients of >0.993. The presented method provides the basis for determination and identification of EtG in human serum and urine samples in a low-concentration range for monitoring alcohol consumption during treatment for alcohol dependence and comorbid alcohol abuse of psychotherapy patients. PMID- 11486834 TI - Systematic analysis of acid, neutral and basic drugs in horse plasma by combination of solid-phase extraction, non-aqueous partitioning and gas chromatography-mass spectrometry. AB - A sample preparation method for mass chromatographic detection of doping drugs from horse plasma is described. Bond Elut Certify (1 g/6 ml) is used for the extraction of 4 ml of horse plasma. Fractionation is performed with 6 ml of CHCl3 Me2CO (8:2) and 5 ml of 1% TEA-MeOH according to its property. Simple and effective clean-up based on non-aqueous partitioning is adopted to remove co eluted contaminants in both acid and basic fractions. Two kinds of 1-(N,N diisopropylamino)-n-alkanes are co-injected with the sample into the GC-MS system for the calculation of the retention index. Total recoveries of 107 drugs are examined. Some data of post administration plasma are presented. This procedure achieves sufficient recoveries and clean extracts for GC-MS analysis. The method is able to detect ng/ml drug levels in horse plasma. PMID- 11486835 TI - High-performance liquid chromatography method for the quantification of non radiolabelled cinnamic compounds in analytes derived from human skin absorption and metabolism experiments. AB - An isocratic high-performance liquid chromatography method has been developed for the quantification of the skin sensitisers trans-cinnamaldehyde and trans cinnamic alcohol, and their cinnamic metabolites. The relative standard deviations (RSDs) between the gradients of eight sets of standard curves were 2.8, 3.1 and 1.9% for cinnamic alcohol, cinnamaldehyde and cinnamic acid, respectively. Sample analytes were derived from two series of experiments: in vitro full-thickness human skin absorption and metabolism studies and metabolism studies using human skin homogenates, with non-radiolabelled cinnamic compounds. Skin absorption and metabolism experiments were performed in the absence and presence of the alcohol dehydrogenase inhibitor, pyrazole. Samples from full thickness skin absorption studies were analysed without extraction; cinnamic compounds from within skin were extracted into methanolic solutions using newly developed methods. The intra-assay RSDs ranged from 0.17 to 2.52% for cinnamic alcohol, 0.24 to 9.14% for cinnamaldehyde and 0.26 to 6.43% for cinnamic acid. The inter-assay RSDs for cinnamic alcohol, cinnamaldehyde and cinnamic acid, respectively, as determined from n=20 HPLC runs, were 2.10, 4.16 and 2.26%. PMID- 11486836 TI - Analysis of phospholipid species in human blood using normal-phase liquid chromatography coupled with electrospray ionization ion-trap tandem mass spectrometry. AB - A narrow-bore normal-phase high-performance liquid chromatography (HPLC) method was developed for separation of phospholipid classes in human blood. The separation was obtained using an HPLC diol column and a gradient of chloroform and methanol with 0.1% formic acid, titrated to pH 5.3 with ammonia and added 0.05% triethylamine. The HPLC system was coupled on-line with an electrospray ionisation ion-trap mass spectrometer. Chromatographic baseline separation was obtained between phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, lyso-phosphatidylcholine, phosphatidylinositol and phosphatidylserine, eluting in that order. The total run time was 30 min. Plasmalogen phosphatidylethanolamine and sphingomyelin, which both are substances with structural similarities to the glycerophospholipids, had similar retention time as phosphatidylethanolamine, but were well separated from the other glycerophospholipid classes. The species from each class were identified using MS2 or MS3, which forms characteristic lyso-fragments. The combination of lyso fragment mass, molecular ion and chromatographic retention time was used to identify each species, including 20 species of phosphatidylglycerol. The mass spectra obtained for the phospholipid classes are presented. Using this system 17 disaturated phospholipid species not earlier described to be present in blood were identified. The limit of detection varied between different phospholipid classes and was in the range 0.1-5 ng of injected substance. PMID- 11486837 TI - Analysis of trace amino acid neurotransmitters in hypothalamus of rats after exhausting exercise using microdialysis. AB - A simple but effective coupling of microdialysis and capillary electrophoresis with laser induced fluorescence detection technique was applied to analysis of amino acid neurotransmitters in the hypothalamus of rats after acute exhausting exercise. The separation of amino acids was achieved using an uncoated fused silica capillary (57 cm x 75 microm I.D.) with a buffer of 10 mM disodium tetraborate at pH 10 and an applied voltage of 12.5 kV. The detection limit was 10(-10) M for each amino acid. It is sufficiently sensitive and rapid for the determination of amino acids in a 5-microl Microdialysate. In comparison to pre exercise, a significant increase in the levels of six hypothalamic amino acids (arginine, glycine, lysine, glutamic acid, alanine, gamma-amino-n-butyric acid) was found after exercise. These results demonstrate that the increase of metabolic amino acids in the hypothalamus of rats can be induced by exhausting exercise and suggests that amino acid neurotransmitters may play functional roles in the central effects of exercise. PMID- 11486838 TI - Simultaneous determination of methionine and total homocysteine in human plasma by gas chromatography-mass spectrometry. AB - A gas chromatographic-mass spectrometric method for the simultaneous determination of methionine and total homocysteine in human plasma is described. DL-[2H4]Methionine and DL-[2H8]homocystine were used as internal standards. The method involved reduction of the disulfide bond with dithiothreitol, purification by cation-exchange chromatography using a BondElut SCX cartridge and derivatization with isobutyl chlorocarbonate in water-ethanol-pyridine. Quantitation was performed by selected-ion monitoring of the quasi-molecular ions of N(O,S)-isobutyloxycarbonyl ethyl ester (IBC-OEt) derivatives for methionine and [2H4]methionine, respectively, and the fragment ions ([M+H-COOisoBu-COOEt]+) for TBC-OEt derivatives for homocysteine and [2H4]homocysteine, respectively. The sensitivity, specificity, accuracy and precision of the method were demonstrated to be satisfactory for measuring concentrations of methionine and total homocysteine in human plasma. PMID- 11486839 TI - Determination of acrylamide and glycidamide in rat plasma by reversed-phase high performance liquid chromatography. AB - Acrylamide is a widely used monomer that produces peripheral neuropathy. It is metabolized to the epoxide, glycidamide, which is also considered to be neurotoxic. A new reversed-phase high-performance liquid chromatography (HPLC) method is described that permits simultaneous determination of acrylamide and glycidamide in rat plasma. Samples were deproteinized with acetonitrile and chromatography was performed using isocratic elution and UV absorption detection. The limits of detection for acrylamide and glycidamide were 0.05 and 0.25 microg/ml in plasma, respectively, and recovery of both analytes was greater than 90%. The assay was linear from 0.1 to 100 microg/ml for acrylamide and from 0.5 to 100 microg/ml for glycidamide. Variation over the range of the standard curve was less than 15%. The method was used to determine the concentration-time profiles of acrylamide and glycidamide in the plasma of acrylamide-treated rats. PMID- 11486840 TI - Direct analysis of urinary trans,trans-muconic acid by coupled column liquid chromatography and spectrophotometric ultraviolet detection: method applicability to human urine. AB - A coupled column liquid chromatographic (LC-LC) method for the direct analysis in human urine of the ring opened benzene metabolite, trans,trans-muconic acid (t,t MA) is described. The method was tested using urine samples collected from five refinery workers exposed to concentrations of airborne benzene (0.2-0.5 ppm), and from non-exposed volunteers. The analytical columns used were of 50 x 4.6 mm I.D. packed with 3 microm p.s. Microspher C18 material as the first column (C-1), and a 100 x 4.6 mm I.D. column packed with 3 microm p.s. Hypersil ODS material as the second one (C-2). The mobile phases applied consisted, respectively, of methanol 0.074% trifluoroacetic acid (TFA) in water (4:96, v/v) on C-1, and of methanol 0.074% TFA in water (10:90, v/v) on C-2. Under these conditions t,t-MA eluted 15 min after injection. The present method, coupling the LC-LC technique with UV detection at 264 nm, permits the quantitation of t,t-MA directly in urine at levels as low as 0.05 mg/l. The determination is performed with a sample throughput of 2 h(-1) requiring only pH adjustment and centrifugation of the sample. Calibration plots of standard additions of t,t-MA to pooled urine taken from five non-exposed subjects were linear (r>0.999) over a wide concentration range (0.05, 0.1, 0.5, 1.0, 2.0 mg/l). The precision of the method (RSD) was in the range of 0.5 to 3.8%, and the within-session repeatability on workers urine samples (levels 0.06, 0.1, 0.2, 1.0 mg/l) was in the range of 3 to 8%. The present method improves the applicability of routine t,t-MA analysis, where it is most desirable that a large number of biological samples can be processed automatically or with minimal human labour, at low cost, and with a convenient turn-around time. PMID- 11486841 TI - Determination of amlodipine in human plasma by high-performance liquid chromatography with fluorescence detection. AB - A sensitive and specific HPLC method has been developed for the assay of amlodipine in human plasma. The assay involves derivatization with 4-chloro-7 nitrobenzofurazan (NBD-Cl), solid-phase extraction on a silica column and isocratic reversed-phase chromatography with fluorescence detection. Nortriptyline hydrochloride was used as an internal standard. The assay was linear over the concentration range of 0.25-18.00 ng/ml. Both of the within-day and day-to-day reproducibility and accuracy were less than 11.80% and 12.00%, respectively. The plasma profile following a single administration of 10 mg amlodipine to a healthy volunteer was presented. PMID- 11486842 TI - Determination of ephedrines in urine by gas chromatography-mass spectrometry. AB - A selective gas-liquid chromatographic method with mass spectrometry (GC-MS) for the simultaneous confirmation and quantification of ephedrine, pseudo-ephedrine, nor-ephedrine, nor-pseudoephedrine, which are pairs of diastereoisomeric sympathomimetic amines, and methyl-ephedrine was developed for doping control analysis in urine samples. O-Trimethylsilylated and N-mono-trifluoroacetylated derivatives of ephedrines--one derivative was formed for each ephedrine--were prepared and analyzed by GC-MS, after alkaline extraction of urine and evaporation of the organic phase, using d3-ephedrine as internal standard. Calibration curves, with r2>0.98, ranged from 3.0 to 50 microg/ml depending on the analyte. Validation data (specificity, % RSD, accuracy, and recovery) are also presented. PMID- 11486843 TI - Rapid high-performance liquid chromatographic method for the simultaneous determination of retinol, alpha-tocopherol and beta-carotene in human plasma and low-density lipoproteins. AB - A reversed-phase HPLC method with diode-array detection was used to simultaneously determine retinol, alpha-tocopherol and beta-carotene in human plasma and low-density lipoproteins. An aliquot of sample was de-proteinized with ethanol containing beta-tocopherol acetate as internal standard, and the analytes were extracted twice with hexane. The solvent was evaporated to dryness under a stream of nitrogen and the residue was redissolved in methanol to be injected directly into the HPLC system. A multiple solvent system based on methanol, butanol and water at a flow-rate of 2 ml/min and held at 45 degrees C provided clear separation of these compounds in only 8 min. The method showed good linearity, precision and accuracy for all compounds. Owing to its simplicity, this method may be useful in routine clinical and epidemiological work. PMID- 11486844 TI - Fast analysis of antibacterial isothiazolones by capillary electrophoresis. AB - Some technical aspects influencing the total time of CE analysis are discussed. A high throughput electrophoretic system based on micellar electrokinetic chromatography (MEKC) is demonstrated as an example. A short capillary, strong electric field, alkaline buffer (pH 9.5) generating strong electroosmotic flow, and parallel hydrodynamic pressure allow the separation of two uncharged isothiazolone derivatives within 45 s. PMID- 11486845 TI - Oxygenator thrombosis: worst case after development of an abnormal pressure gradient--incidence and pathway. AB - The development of an abnormally high pressure gradient (APG) before the membrane oxygenator (MO) is a complication that occurs during some extracorporeal circulation (ECC) procedures. The present study deals with the incidence of an APG and discusses a probable causative pathway by comparing surface-coated and uncoated oxygenation systems. Five thousand six hundred and seventeen adult ECCs were carried out (2,581 without and 3,036 with surface coatings). The incidence of an APG, therefore, amounted to 0.03% in the group with coated systems and 4.3% in the uncoated group. In addition, an in vitro study demonstrated significantly reduced adhesion and activation of platelets and leucocytes when the surfaces of the MOs were coated with heparin or polypeptides. The advantages of coating surfaces of ECC devices possibly depend on the selective adsorption of particular plasma proteins. These will presumably form a biocompatible membrane on the surface, and minimize pathological deposit of fibrin, platelets and other blood cells, and, therefore, implicate the prevention of an oxygenator failure. PMID- 11486846 TI - Effect of Carmeda BioActive Surface coating versus Trillium Biopassive Surface coating of the oxygenator on circulating platelet count drop during cardiopulmonary bypass. AB - An investigation was conducted to evaluate the effect that surface coating of the hollow-fiber membrane oxygenator had on circulating platelet count drop during cardiopulmonary bypass (CPB). Sixty patients undergoing non-emergency myocardial revascularization for coronary artery disease were randomly divided into two groups. Group one (n = 32) received the Carmeda-coated Maxima-Plus PRF oxygenator while the patients in Group two (n=28) received the Trillium-coated Affinity oxygenator during CPB. The net platelet count drops for the pump specimen (15-20 min after the initiation of bypass) for the Carmeda and the Trillium groups were 3.6 +/- 15.8% and 6.2 +/- 10.2%, respectively. The net platelet count drop for the warming specimen for the Carmeda and the Trillium groups were 2.9 +/- 19.4% and 0.5 +/- 11.0%, respectively. There were no statistically significant differences between the groups. The authors conclude that using either the Carmeda-coated Maxima-Plus PRF oxygenator or the Trillium-coated Affinity oxygenator afford similar benefits in regards to preserving circulating platelet counts during bypass. PMID- 11486847 TI - Veno-venous bypass in liver transplantation: heparin-coated perfusion circuits reduce the activation of humoral defense systems in an in vitro model. AB - We studied the effects of bypass circuit surface heparinization on kallikrein kinin, coagulation, fibrinolytic and complement activation in a closed model system for simulating veno-venous bypass (WBP) in orthotopic liver transplantation (OLT). The circuits were identical to those in routine use during clinical OLT in our institution. Fresh whole human blood diluted 1:2 with Ringer's acetate was circulated at a non-pulsatile flow (2 l/min) and at a constant temperature (37.5 degrees C) for 12 h. In 10 experiments, the entire inner surface of the circuits was coated with end-point attached heparin (HC). In the remaining 10, non-treated PVC tubing was used (NC). Components of the plasma kallikrein-kinin, coagulation, fibrinolytic and complement systems were analyzed using functional techniques (chromogenic peptide substrate assays) and enzyme immunoassays at baseline, 3 and 12 h. Significant activation of the initial (C3bc) and terminal (TCC) components of the complement system were found in both the NC and HC groups after 3 and 12 h: C3bc: NC: baseline = 4 (3.5-7.7), 3 h = 17.3* (12.5-27), 12h = 31* (17.7-63.6), HC: baseline = 4.9 (3.2-6.8), 3h = 9* (6 14.4), 12h = 13.7* (7.4-18.1). TCC: NC: baseline = 0.4 (0.2-0.6), 3h = 5*(0.8 11.9), 12 h: 13.1* (4.2-25.7). HC: baseline = 0.5 (0.1-0.6), 3 h = 0.6* (0.1 0.8), 12 h = 1.2* (0.3-2) AU/ml; median and range (*: p < 0.05). The C3bc and TCC concentrations were significantly higher in the NC group at 3 and 12 h, compared to the HC group: C3bc (NC vs. HC group): 3 h, p < 0.001; 12 h, p < 0.001. TCC (NC vs. HC group): 3h, p < 0.001; 12 h, p < 0.001. Significant increases in the values of thrombin-antithrombin complexes (p = 0.003), prothrombin fragment 1 + 2 (p = 0.006) and plasmin-alpha2-antiplasmin complexes (p = 0.016) were found in the non-coated group, but not in the heparin-coated group during the observation period, showing that the coagulation and fibrinolytic systems were activated in the non-coated circuits. We conclude that heparin-coating of the internal surface of the extracorporeal perfusion circuit used for WBP reduces activation of the plasma cascade systems in a closed venous system in vitro. PMID- 11486848 TI - An in vitro study of the effects of isoflurane on oxygen transfer. AB - A common anesthetic technique utilized during cardiopulmonary bypass (CPB) includes the use of various inhalation agents, such as isoflurane. The purpose of this study was to evaluate the effects of this agent on oxygen transfer during CPB. An in vitro model was designed using bovine blood. Blood flow was held constant at 2 l/min, while gas flow was manipulated at 1 and 3 l/min. The percentage of inspired oxygen (FiO2) was set at 50 and 100%, and isoflurane was manipulated to 1.0, 3.0 and 5.0%. Blood gas analysis, oxygen transfer, and inlet and outlet isoflurane concentrations were measured at each of the given conditions. A total of 12 trials with four oxygenators were conducted. In the four oxygenators used in our study, no significant differences in oxygenator performance were found. At conditions of 1 I/min gas flow, 50% FiO2 and 1% isoflurane, there were no significant changes in O2 transfer between baseline and measurements taken during isoflurane administration (100.18 +/- 12.49 vs 102.35 +/- 10.99 ml O2/min, p=0.8031). At 3 I/min gas flow, 100% FiO2 and 5% isoflurane, no significant differences were found (142.35 +/- 10.76 vs 154.04 +/- 8.95 ml O2/min, p=0.1459). The only significant differences found for oxygen transfer were between 50 and 100% FiO2, all other conditions being set equal (102.35 +/- 10.99 vs 137.68 +/- 8.62 ml O2/min, p=0.0023). In conclusion, increasing concentrations of isoflurane up to 5% does not affect the efficiency of oxygen transfer in an in vitro circuit. Further studies are necessary to evaluate the effects in an in vivo setting. PMID- 11486849 TI - Is postoperative blood loss, loss of blood? A pilot study in cardiac surgical patients. AB - The effect of estimating the blood balance using changes in erythrocyte volumes (EVs) instead of the routinely used changes in haematocrit values was studied in 20 patients scheduled for cardiac surgery. We determined the mean haematocrit of the effluent from the postoperative thoracic drainage system at various time intervals. These data were used to more accurately calculate the blood balance. From 8h after surgery onwards, the haematocrit in the thoracic effluent was less than 10%. Total loss of thoracic effluent until 24h after removal of the aortic crossclamp (ACC) was 1,735 +/- 803 ml. Calculated blood loss until 24 h after ACC was only 58% of the total thoracic effluent. Plasma volumes in these patients increased from preoperative values of 2,505 +/- 499ml at admission to the hospital to maximum levels of 4,969 +/- 1,027 ml at 12 h after ACC (p < 0.05). Blood volume rose to 159% of the preoperative value at 12 h after ACC, whereas the EV remained relatively stable, decreasing to 95% of the preoperative value at 4 h after ACC and increasing to 107% of the baseline value at 24 h after ACC. In the meantime, patient haematocrit decreased to 78% of the reference value at the time of induction of anaesthesia at 4 h after ACC and then increased to 84% at 24 h after ACC. Thus, the use of patient haematocrit considerably overestimates blood loss. The EV appears to be a more appropiate variable than haematocrit in monitoring the blood balance in cardiac surgical patients. Future studies should reveal whether the EV is practicable in daily clinical practice. PMID- 11486850 TI - Centrifugation of salvaged pump blood is useful in reducing the amount of heparin re-infused into the patient. AB - Conservation of blood and blood products are of major concern in clinical practice today. All blood salvaged from the extracorporeal circuit at the end of cardiopulmonary bypass should be returned to the patient. We propose the use of a simple and inexpensive centrifuge machine to reduce the amount of heparin from the residual pump blood re-infused to the patient. At the same time, the re infused blood is concentrated. Sixteen patients were prospectively included in this study. The residual pump blood was collected from the bypass circuit and spun at 4,000 rpm for 7 min on the Jouan GR 4.22 machine. The clear upper supernatant was discarded and the remaining concentrated red cells re-infused back to the patient. The heparin concentration and full blood counts of both layers were measured. We found that this method was useful in removing excessive heparin and avoiding the need for additional protamine. PMID- 11486852 TI - The impact of normothermia on the outcome of aortic valve surgery. AB - The purpose of this study was to examine the effects of systemic perfusion temperature on the clinical outcome after aortic valve surgery. In this study, we examined 323 patients who underwent aortic valve surgery between January 1994 and April 1996. Forty-six patients were perfused in moderate hypothermia (28 degrees C) and 277 patients in normothermia. Age and sex distribution of the patients were similar. There were no statistically significant differences between the groups regarding neurological, renal or cardiac complications. Patients in hypothermia required less catecholamine at the end of the operation (p = 0.00001), but there was no significant difference in the length of the stay in the intensive care unit between the groups. Cardiopulmonary bypass temperature did not influence early outcome after aortic valve surgery. PMID- 11486851 TI - Haemolysis due to active venous drainage during cardiopulmonary bypass: comparison of two different techniques. AB - To facilitate mini-access for cardiac surgery, two different methods of active venous drainage are used: vacuum assisted drainage and centrifugal pump aspiration on the venous line. The aim of this study was to compare the haemolysis produced using these two techniques. From June to December 1999, 50 consecutive patients were operated on using a ministernotomy. All of these patients had valvular surgery for either valve repair or valve replacement (9 MVRepair, 11 MVR, 29 AVR, 1 AVR + MVR). They were randomized into two groups: Group A, 25 patients who underwent surgery where vacuum assisted drainage was used, and Group B, 25 patients where kinetic asssisted venous drainage with centrifugal pump venous aspiration was used. Patient characteristics of both groups were similar for age, gender, body weight, body surface area, height, cardiopulmonary bypass (CPB) time, aortic crossclamp time, priming volume, cardioplegia volume, haemoglobin concentration, haematocrit, serum creatinine, bilirubin, lactate dehydrogenase (LDH), serum glutamic oxaloacetic transaminase (sGOT), serum glutamic pyruvic transaminase (sGPT), aptoglobin, reticulocytes, and platelet count. We checked all these laboratory parameters preoperatively, at the end of CPB, and 2 and 24 h after operation. We also checked haemoglobinuria at these same time points. We assessed blood loss at 6, 12, and 24 h after the operation and calculated total postoperative bleeding. There was a tendency towards a greater increase in LDH, sGOT and sGPT in Group A more than in Group B, but these data did not reach statistical significance. Platelet count was always lower in Group A and aptoglobin increased in Group A more than in Group B. More patients in Group A had haemoglobinuria. These findings indicate that haemolysis is increased more in patients treated with vacuum assisted drainage, when compared to the rise in haemolysis in those treated with centrifugal pump venous drainage. Total bleeding is also greater in Group A. PMID- 11486853 TI - Circuit and protocol for hypoxic hyperthermic isolated limb perfusion to treat malignant melanoma. AB - In the treatment of cancer, isolated limb perfusion (ILP) allows what would be a lethal systemic dose of cytotoxic drugs to be administered directly to a tumour site of an extremity. Unfortunately, ILP is a complex, expensive, time-consuming treatment that requires general anaesthesia, vascular surgery and expertise with extracorporeal circuits that may not be available outside a cardiac centre. By streamlining the traditional ILP protocols and eliminating the oxygenator from the circuit, an equally safe and effective technique of hypoxic hyperthermic isolated limb perfusion has been developed. PMID- 11486854 TI - Computer simulation of ion chromatography separation: an algorithm enabling continuous monitoring of anion distribution on an ion-exchange chromatography column. AB - A computer algorithm for the calculation of ion chromatography separation is presented. It is based on the calculation of equilibrium concentrations of present analyte in discrete column segments. The continuous column is treated as a number of discrete cells or segments where the equilibration process between the stationary phase and the eluent is simulated. The ion-exchange equilibration process is supposed to be instantaneous and quantitative. The continuous flow of the eluent is rendered by discrete transfers. The size of each transfer of the eluent corresponds to a portion of the volume contained in one column segment. The equilibrium calculations in all column segments are repeated for each transfer of the eluent, through all the stages of the chromatographic process. The distribution of the analytes between the stationary phase and the eluent can be monitored at any step and in any column segment which means that the described algorithm provides the spatial and time concentration profiles. The simulated chromatogram is acquired as a time-concentration profile in the last column segment. The obtained chromatograms are in good agreement with the experimental ones. The distribution of ions between the stationary phase and the eluent in the early stages of the ion chromatographic process can thus be studied with confidence. PMID- 11486855 TI - Determination of microcystins in blue-green algae, fish and water using liquid chromatography with ultraviolet detection after sample clean-up employing immunoaffinity chromatography. AB - Anti-microcystin LR immunnoaffinity cartridges were evaluated for their ability to selectively remove microcystins from extracts of blue-green algae, fish and water samples for subsequent analysis by liquid chromatography with UV absorbance detection at 238 nm. Blue-green algae and fish samples were extracted with 75% methanol in water. A portion of the extract was diluted and passed through an immunoaffinity cartridge. Water samples were applied directly to the cartridge. The cartridge was rinsed with water and 25% methanol in water. The microcystins were eluted with 80% methanol in water containing 4% acetic acid. It was found that the cartridges were effective in isolating the microcystins from blue-green algae, fish and water samples, resulting in extracts that were clean enough to enable direct LC-UV detection down to approximately 0.03 microg/g in the blue green algae and fish samples, and as low as 0.02 ng/ml for water samples. The cartridges were found to have a capacity of approximately 200 ng each for a mixture of microcystins RR, YR, LR and LA, or as much as 525-800 ng for individual compounds. Recoveries trough the complete analytical procedure ranged from 64 to 115% (all values) with an overall average of approximately 80% at spiking levels of 0.5-4.0 microg/g for the microcystins in blue-green algae. The average recoveries (n=8) from spiked (0.1-0.5 microg/g) fish samples were 73% for RR, 79% for YR, 81% for LR and 77% for LA, while from the spiked (2.0-0.04 ng/g) tap and river water samples (n=6), recoveries were 78% for RR, 86% for YR, 94% for LR and 89% for LA. PMID- 11486856 TI - Note on the use of reciprocity of chiral recognition in designing liquid chromatographic chiral stationary phases. AB - A new high-performance liquid chromatographic chiral stationary phase (CSP) was prepared from (S)-N-(3,5-dimethylbenzoyl)leucine N-phenyl N-allyl amide. The new CSP was applied for the resolution of N-(3,5-dinitrobenzoyl)-alpha-amino amides and esters and the chromatographic resolution results were compared with those on another CSP derived from (S)-N-(3,5-dimethoxylbenzoyl)leucine N-phenyl N-allyl amide. The new CSP was found to exert greater enantioselectivity than the other one. These results are contrary to what was expected from the reciprocity of chiral recognition. From these results it was concluded that the reciprocity of chiral recognition should be used with some degree of care in developing effective CSPs or in predicting chromatographic resolution behaviors. PMID- 11486857 TI - Enantioseparation of selected chiral sulfoxides using polysaccharide-type chiral stationary phases and polar organic, polar aqueous-organic and normal-phase eluents. AB - HPLC enantioseparation of selected chiral sulfoxides was studied using cellulose and amylose phenylcarbamate derivatives as chiral stationary phases (CSPs). The contributions of various functional groups of a chiral analyte as well as the polysaccharide derivatives in the analyte retention and chiral recognition were evaluated. A very high enantioseparation factor exceeding 110 was observed in the enantioseparation of 2-(benzylsulfinyl)benzamide (BSBA) on cellulose tris(3,5 dichlorophenylcarbamate) (CDCPC) CSP by using 2-propanol as a mobile phase. The enantiomer elution order was opposite on cellulose and amylose phenylcarbamates. For the polysaccharide-type CSPs, pure alcohols such as methanol, ethanol and 2 propanol represent a valuable alternative to more common alcohol-hydrocarbon and reversed-phase eluents. PMID- 11486858 TI - Quantitative structure-enantioselective retention relationships for chromatographic separation of arylalkylcarbinols on Pirkle type chiral stationary phases. AB - Quantitative structure-retention (QSRR, retention factors log k1 and log k2 for the first and second eluted enantiomer) as well as enantioselective retention relationships (QSERR, separation factor log a) for a series of 42 chiral arylalkylcarbinols on four brush-type chiral stationary phases are derived by multiple linear regression analyses and artificial neuronal network calculations using 2D and 3D molecular descriptors including those obtained by quantum chemical calculations. Separation factors are in addition modeled by the 3D-QSAR method of comparative molecular field analysis (CoMFA). For the retention factors the LUMO energy turns out to be the most important descriptor, whereas for log a it is the hydrophobicity of the analytes. With CoMFA both the steric and electrostatic field are found to be of almost comparable significance. PMID- 11486859 TI - Union of capillary high-performance liquid chromatography and microcoil nuclear magnetic resonance spectroscopy applied to the separation and identification of terpenoids. AB - This paper describes the first coupling of a commercial capillary HPLC system with a diode array spectrophotometric detector and a custom-built nuclear magnetic resonance (NMR) flow microprobe. The eluent from a 3-microm diameter C18 HPLC column is linked to a 500 MHz 1H-NMR microcoil probe with an observe volume of 1.1 microl. The separation and structurally-rich detection of a mixture of terpenoids under both isocratic and gradient solvent elution conditions is presented. The lowest limits of detection yet reported for capillary HPLC on-line measurement (i.e., 37 ng for alpha-pinene) are achieved with this system. The complementary nature of diode array and NMR detection allows stopped-flow data collection from analytes which would otherwise go unnoticed in continuous-flow NMR. Moreover, stopped-flow NMR data is presented for the detection of a trace (sub-nmol) impurity in the sample mixture. Since NMR signals degrade and shift during solvent gradients, flow injection analysis studies are conducted with injected solvent plugs differing in mobile phase composition. The NMR signal degradation accompanying these injections is largely due to the variance in chemical shift with the solvent composition rather than to changes in magnetic susceptibility of the solvent. Characterization of such effects enables the development of improved NMR probes for the coupling of capillary HPLC and NMR. PMID- 11486860 TI - Chiral separation of amino acids by copper(II) complexes of tetradentate diaminodiamido-type ligands added to the eluent in reversed-phase high performance liquid chromatography: a ligand exchange mechanism. AB - In this paper we report a study on the mechanism of the enantiomeric separation of unmodified D,L-amino acids in RP-HPLC by copper(II) complexes of two tetradentate diaminodiamido ligands, (S,S)-N,N'-bis(phenylalanyl)ethanediamine (PheNN-2) and (S,S)-N,N'-bis(methylphenylalanyl)ethanediamine (Me2PheNN-2), added to the eluent. The aim is to investigate whether and how a copper(II) complex with no free equatorial positions can perform chiral discrimination of bidentate analytes such as unmodified amino acids. The problem is approached in a systematic way by: (a) varying the different chromatographic parameters (pH, selector concentration, eluent polarity); (b) performing chiral separation with the selector adsorbed on the stationary phase; (c) studying the ternary complex formation of these ligands with D- and L-amino acids in solution by glass electrode potentiometry and electrospray ionization MS. All the experimental data are consistent with a mechanism of chiral recognition, based on ligand exchange, which involves as selectors the species [Cu2L2H(-2)]2+ and [CuLH(-2)] and proceeds by displacement of two binding sites from the equatorial positions, giving rise to the ternary species [CuLA]+ and [CuLH(-1) A]. The most important factor responsible for chiral discrimination seems to be the affinity of the diastereomeric ternary complexes for the stationary phase since no enantioselectivity is observed in solution. PMID- 11486861 TI - Ni(II)-based immobilized metal ion affinity chromatography of recombinant human prolactin from periplasmic Escherichia coli extracts. AB - A novel, two-step preparative technique is described for the purification of authentic recombinant human prolactin (rhPRL) secreted into the periplasm of transformed Escherichia coli cells. The first step is based on immobilized metal ion affinity chromatography of periplasmic extract, using Ni(II) as a relatively specific ligand for hPRL in this system. It gives superior resolution and yield than established ion-exchange chromatography. Size-exclusion chromatography is used for further purification to >99.5% purity. The methodology is reproducible, leading to 77% recovery. Identity and purity of the rhPRL were demonstrated using sodium dodecylsulphate-polyacrylamide electrophoresis, isoelectric focusing, mass spectrometry (matrix-assisted laser desorption ionization time-of-flight), radioimmunoassay, RP-HPLC and high-performance size-exclusion chromatography. In the Nb2 bioassay, the hormone showed a bioactivity of 40.9 IU/mg. PMID- 11486862 TI - Recognition and strand displacement of DNA oligonucleotides by peptide nucleic acids (PNAs). High-performance ion-exchange chromatographic analysis. AB - Peptide nucleic acids (PNAs) are oligonucleotide mimics containing a pseudopeptide chain, which are able to bind complementary DNA tracts with high affinity and selectivity. Two mixed-sequence PNA undecamers (1 and 2) were synthesized and their double-stranded adducts with the complementary oligonucleotides (3 and 4) were revealed by the appearance of the corresponding peak in anion-exchange HPLC. A DEAE column was used and elution was performed with aqueous Tris buffer (pH 8) and an ionic strength gradient (0-0.5 M NaCl). The same effect was not observed with non-complementary oligonucleotides. The stability of the PNA-DNA adducts under the conditions used in the chromatographic system was studied as a function of temperature. Furthermore, in competition experiments double-stranded oligonucleotides were challenged by a PNA complementary to one strand: the formation of the PNA-DNA hybrid and the displacement of the non-complementary strand were observed with high specificity. The results suggest a possible use of ion-exchange HPLC for studying PNA-DNA interactions, and indicate the efficiency of PNA probes in the chromatographic analysis of DNA. PMID- 11486863 TI - Application of mixed partition-adsorption systems in high-performance liquid chromatography of purines and pyrimidines. AB - Separation of the test mixtures of some purine and pyrimidine derivatives on silicas (types A and B) in adsorption normal-phase (A-NP) and mixed partition adsorption normal-phase (MPA-NP) mode has been studied. When the A-NP mode is used the peak shapes are unsatisfactory (especially on type A silica). At the same time MPA-NP systems show a good peak symmetry on all silica types. The findings have demonstrated that the MPA-NP mode offers a specific selectivity. This allows MPA-NP systems to supersede reversed-phase systems in some application areas. PMID- 11486864 TI - Liquid exclusion adsorption chromatography, a new technique for isocratic separation of nonionic surfactants. III. Two-dimensional separation of fatty alcohol ethoxylates. AB - A quantitatively accurate mapping of lower fatty alcohol ethoxylates can be achieved using a combination of liquid chromatography under critical conditions as the first dimension and liquid exclusion-adsorption chromatography as the second dimension. With coupled density and refractive index detection in both dimensions, the contribution of preferential solvation can also be estimated. In most cases, however, the use of refractive index detection alone also yields satisfactory results. PMID- 11486865 TI - Evaluation of tetraglyme for the enrichment and analysis of volatile organic compounds in air. AB - A recently developed method for the sampling and analysis of volatile organic compounds in air has been evaluated. The system is based on the enrichment of analytes in tetraethylene glycol dimethyl ether or tetraglyme, a water-soluble organic liquid. The subsequent analysis consists of dispersion of a sample aliquot in water followed by purge-and-trap and gas chromatographic separation. Physico-chemical data were investigated for 10 volatile organic compounds, providing information on the possibilities and limitations of the tetraglyme method. The target analytes included chlorinated alkanes and alkenes, and monocyclic aromatic hydrocarbons. Air/tetraglyme partition coefficients Kat were determined over an environmental relevant temperature range of 2-25 degrees C to evaluate sorption efficiencies and estimate breakthrough volumes at the sampling stage. At 2 degrees C breakthrough volumes (allowing 5% of breakthrough) ranged from 5.8 (1,1-dichloroethane) to 312 l (1,1,2-trichloroethane) for 20 ml of tetraglyme. With regard to the desorption stage, the effect of tetraglyme on the air/water partition of organic compounds was investigated through the measurement of air/tetraglyme-water partition coefficients Kat-w for 2-31% (v/v) tetraglyme in water. Finally a clean-up procedure for tetraglyme was evaluated. Analysis of a blank tetraglyme-water (17:83, v:v) mixture by gas chromatography-flame ionization detection/mass spectrometry showed minor background signals. None of the target compounds were detected. PMID- 11486866 TI - Chromatographic method for diaminopimelic acid detection in calcareous rocks. Presence of a bacterial biomarker in stromatolites. AB - The presence in the environment of diaminopimelic acid (DAP), a specific eubacterial marker, can be attributed to that of bacteria. We report a reliable and highly sensitive method for the quantification of DAP in calcareous rocks. It consists of acid hydrolysis of rock powder, purification of DAP by chromatography on Dowex 50W and Spherogel AA-NA+ columns, and quantitative analysis by high performance liquid chromatography. Addition of tritiated DAP, the internal standard, allows one to follow the relevant fractions throughout the purification procedure and to determine their yield. The analytical step consists in pre column derivatization with ortho-phthaldialdehyde of purified samples, and separation through a reversed-phase C18 column. Chemical controls, i.e., oxidation of samples to rule out the presence of co-eluting lanthionine and cystathionine, as well as mass spectrometry, confirm the presence of DAP in analyzed samples. Our method allows the separation of meso- from L- and/or D stereoisomers of DAP, and reveals their presence in the examined rocks, two stromatolites of different age and geographic origin. PMID- 11486867 TI - Gas chromatography-mass spectrometry method for the simultaneous determination of wood extractive compounds in quaking aspen. AB - We have developed a rapid gas chromatography-mass spectrometry (GC-MS) method for the detailed compositional analysis of 70 underivatized wood extractive components present in quaking aspen (Populus tremuloides Michx.). Forty-four compounds were unequivocally identified by retention time and mass spectral comparison with standards. An additional 26 chromatographic peaks were assigned to broad chemical classes using retention time and mass spectra features. The results were compared to the respective tert.-butyldimethylsilyl derivatized wood extractives profile, and it was determined that derivatization was unnecessary for the GC-MS analysis of the target compounds. PMID- 11486868 TI - Gas chromatographic-mass spectrometric analysis of products arising from pyrolysis of amino acids in the presence of hexamethyldisilazane. AB - Alpha-amino acids were pyrolysed at 600 degrees C in the presence of hexamethyldisilazane (HMDS) and the formed volatile products were analysed on line by gas chromatography-mass spectrometry (GC-MS). Glycine, alanine, valine, leucine, isoleucine, norleucine, methionine, phenylalanine yielded principally the trimethylsilyl (TMS) ester of the parent amino acid. TMS esters of carboxylic acids arising from reductive deamination were observed for serine, threonine and aspartic acid. Decarboxylation resulted in the formation of amines which represented abundant products released from tyrosine, cysteine and methionine. Cyclic compounds arising from the condensation of two amino acids were revealed as characteristic products of glycine, alanine, serine, proline and hydroxyproline. Degradation products of the side chain were released at relatively high levels from tryptophane, tyrosine and hystidine. Since each amino acid produced a characteristic distribution of TMS products, in-situ pyrolysis/silylation with HMDS may find application as a screening technique for the detection of amino acids and related materials in complex matrices. The potentiality of the procedure was tested on a dipeptide (Tyr-Leu). PMID- 11486869 TI - Optimization of headspace solid-phase microextraction conditions for the determination of organophosphorus insecticides in natural waters. AB - Headspace solid-phase microextraction (HS-SPME) has been developed for the analysis of seven organophosphorus insecticides, i.e. diazinon, fenitrothion, fenthion, ethyl parathion, methyl bromophos, ethyl bromophos and ethion in natural waters. Their determination was carried out using gas chromatography with flame thermionic and mass spectrometric detection. To perform the HS-SPME, two types of fibre have been assayed and compared: polyacrylate (PA 85 microm), and polydimethylsiloxane (PDMS 100 microm). The main parameters affecting the HS-SPME process such as temperature, salt additives, memory effect, stirring rate and adsorption-time profile were studied. The method was developed using spiked natural waters such as ground, sea, river and lake water in a concentration range of 0.05-1 microg/l. The HS-SPME conditions were optimized in order to obtain the maximum sensitivity. Detection limits varied from 0.01 to 0.04 microg/l and relative standard deviations (RSD <17%) were obtained showing that the precision of the method is reliable. The method showed also good linearity for the tested concentration range with regression coefficients ranging between 0.985 and 0.999. Recoveries were in relatively high levels for all the analytes and ranged from 80 to 120%. Water samples collected from different stations along the flow of Kalamas river (NW Greece) were analyzed using the optimized conditions in order to evaluate the potential of the proposed method to the trace-level screening determination of organophosphorus insecticides. The analysis with HS-SPME has less background interference and the advantage of its non-destructive nature reveal the possibility of the repetitive use of the SPME fibre. PMID- 11486870 TI - Estimation of width of narrow molecular-weight distributions by size-exclusion chromatography with concentration and light scattering detection. AB - A new method for the estimation of the weight-to-number-average molecular-weight ratio, Mw/Mn of polymers with a narrow molecular-weight distribution, approximated by log-normal distribution, is proposed using size-exclusion chromatography (SEC) with concentration and light-scattering detectors. From experimental data, the Mw/Mn ratios are calculated by two procedures: one using the concentration and light-scattering elution curve for the polymer measured, and the other based on the concentration elution curve and calibration line for a wide range of molecular masses. An iteration method has been developed making the two Mw/Mn ratios converge. The method was applied to a series of narrow molecular weight distribution polystyrene standards. PMID- 11486871 TI - Determination of organophosphorus pesticides in honeybees after solid-phase microextraction. AB - A method based on solid-phase microextraction (SPME) followed by gas chromatography with nitrogen-phosphorus detection was developed for the purpose of determining 18 organophosphorus pesticide residues in honeybee samples (Apis mellifera). The extraction capacities of polyacrylate and poly(dimethylsiloxane) fibers were compared. The main factors affecting the SPME process, such as the absorption time profile, salt, and temperature, were optimized. The method involved honeybee sample homogenization, elution with an acetone:water solution (1:1) and dilution in water prior to fiber extraction. Moreover, the matrix effect on the extraction was evaluated. In samples spiked at the 0.2 mg kg(-1) level, the coefficient variation was between 1 and 13% and the detection limits were below 10 microg kg(-1). The SPME procedure was found to be quicker and more cost-effective than the solvent extraction method commonly used. The method was applied successfully to environmental screening. Parathion methyl was detected and confirmed in the real samples analyzed. PMID- 11486872 TI - Polydimethylsiloxane solid-phase microextraction-gas chromatography method for the analysis of volatile compounds in wines. Its application to the characterization of varietal wines. AB - A study was made of the validity of the solid-phase microextraction method, using a polydimethylsiloxane coated fused-silica fiber, for the extraction-desorption of the minor volatile compounds from wine before their gas chromatographic analysis. The aspects considered were the influence of ethanol on extraction, repeatability, limits of detection, linearity and recovery of compounds. This method, together with the direct injection of the major volatile compounds, was applied to 16 varietal wines. The findings indicate that the method is a highly suitable technique for the analysis of wines and that the volatile composition of wines depends, at least partly, on the grapes with which they have been made. PMID- 11486873 TI - Migration of neutral solutes by double stepwise gradient elution in capillary electrochromatography. AB - Characteristics of electroosmotic flow (EOF) and the migration of neutral solutes under double stepwise gradient elution in capillary electrochromatography were studied systematically. EOF velocity proved to be the function of operation time changing with the introduction of the second mobile phase. Accordingly, the retention of components also changed. The migration of neutral solutes was studied under the following three situations; A, components eluted when the column was filled only with the first kind of mobile phase; B, solutes eluted still in the first kind of mobile phase while at that time two kinds of mobile phase coexisted in the column and C, samples eluted in the second kind of mobile phase. Equations to describe the retention times of components under these three kinds of conditions were deduced and applied to predict the retention times of 12 aromatic compounds. Relative errors between experimental and calculated values were below 5.0%, which proved the reliability of the equations. In addition, parameters that might affect the retention time of solutes, such as the transferring time of mobile phase vials, the capacity factors of components and EOF velocities two steps were studied systematically. PMID- 11486874 TI - Experimental investigation on moving chemical reaction boundary theory for weak acid-strong-base system with background electrolyte KCl in large concentration. AB - In this report, the moving chemical reaction boundary (MCRB) was formed with the weak acid of acetic acid (HAc) and the strong alkali of NaOH, coupled with the excess of background electrolyte KCl. The experiments were compared with the predictions by the moving chemical reaction boundary equation (MCRBE). It is very interesting that (1) the experimental results are in good agreement with the predictions with the original MCRBE if the MCRB is an anodic moving boundary, (2) however, the experiments are extremely far away from the predictions with the original MCRBE if a cathodic moving boundary. Hence, the original MCRBE must be corrected under the later situation of cathodic moving MCRB. The corrected MCRBE was well quantitatively proved to be valid for the cathodic moving MNRB formed with the same electrolytes of HAc, NaOH and KCl. PMID- 11486875 TI - Separation of hydrophobic polymer additives by microemulsion electrokinetic chromatography. AB - Microemulsion electrokinetic chromatography (MEEKC) has been applied to the separation of some phenolic antioxidants [Irganox 1024, Irganox 1035, Irganox 1076, Irganox 1010, Irganox 1330, Irgafos 138, Irganox 168 and 2,6-di-tert.-butyl 4-methylphenol (BHT)]. Due to the extremely hydrophobic nature of these analytes, they could not be separated using standard MEEKC conditions and two alternative approaches were investigated. Using an acidic buffer (phosphate, pH 2.5) to effectively suppress the electroosmotic flow, the addition of 2-propanol to the aqueous phase of the microemulsion buffer to improve partitioning of the analytes, and a negative separation voltage, separation of five of the analytes in under 10 min was possible. The second approach, using a basic buffer (borate, pH 9.2) and a positive separation voltage resulted in complete resolution of all eight analytes. A mixed surfactant system comprising the anionic sodium dodecyl sulfate (SDS) and neutral Brij 35 was used to reduce the overall charge and with it the mobility of the droplets, and hence the separation time. Using an optimised MEEKC buffer consisting of 2.25% (w/w) SDS, 0.75% (w/w) Brij 35, 0.8% (w/w) n-octane, 6.6% (w/w) 1-butanol, 25% (w/w) 2-propanol and 64.6% (w/w) 10 mM borate buffer (pH 9.2) the eight target analytes were baseline separated in under 25 min. For these analytes, MEEKC was found to be superior to micellar electrokinetic chromatography in every respect. Specifically, the solubility of the analytes was better, the selectivity was more favourable, the analysis time was shorter and the separation efficiency was up to 72% higher when using the MEEKC method. Detection limits from 5.4 to 26 microg/ml were obtained and the calibration plot was linear over more than one order of magnitude. The optimised method could be applied to the determination of Irganox 1330 and Irganox 1010 in polypropylene. PMID- 11486876 TI - Non-aqueous capillary electrophoretic separation of enantiomeric amines with (-) 2,3:4,6-di-O-isopropylidene-2-keto-L-gulonic acid as chiral counter ion. AB - (-)-2,3:4,6-Di-O-isopropylidene-2-keto-L-gulonic acid [(-)-DIKGA] has been introduced as a chiral counter ion in non-aqueous capillary electrophoresis. High enantioresolutions (R(s)> or =3) were obtained for amines, e.g., pronethalol, labetalol and bambuterol. Methanol containing NaOH and (-)-DIKGA was used as the background electrolyte. The counter ion concentration and the nature of the injection medium were found to affect the chiral separation. Covalent coating of the fused-silica capillary reduced the electro-osmotic flow resulting in improved enantioresolutions. PMID- 11486877 TI - Capillary electrophoretic separation of protease inhibitors used in human immunodeficiency virus therapy. AB - The scope of this work was to investigate the migration behavior of the currently used protease inhibitors for antiretroviral therapy of people infected with the human immunodeficiency virus and to develop a method for their capillary electrophoretic separation and determination. All of the protease inhibitors (indinavir, saquinavir, nelfinavir, amprenavir, and ritonavir) contain at least one basic amino functional group. As a consequence, they can be separated by capillary zone electrophoresis using acidic buffer electrolytes. A fast electroosmotic flow is established in order to increase separation speed, by adding a cationic electroosmotic flow modifier to the electrolyte. After using conventional serum pretreatment procedures it is possible to separate all five protease inhibitors within less than 5 min. In addition, a non-aqueous CE method is also presented which enables the separation of three protease inhibitor compounds within less than 3 min. PMID- 11486878 TI - Non-aqueous capillary electrophoresis with diode array and electrospray mass spectrometric detection for the analysis of selected steroidal alkaloids in plant extracts. AB - Nonaqueous capillary electrophoresis coupled to UV detection is described for the separation and determination of steroidal alkaloids. After optimization of electrophoretic parameters, including the electrolyte nature and the organic solvent composition, a reliable separation of solasodine and solanidine was achieved in a methanol-acetonitrile (20:80, v/v) mixture containing 25 mM ammonium acetate and 1 M acetic acid. For quantitative purposes, a fused-silica capillary with a bubble cell was used and detection was performed at low wavelength (195 nm). Method performances, including migration time and peak area reproducibility, linearity, sensitivity and accuracy, were also evaluated. The method was applied to determine solasodine in Solanum elaeagnifolium berries and Solanum sodomaeum leaves and seeds. To further improve sensitivity in the analysis of solasodine-related compounds, solanidine, demissidine and tomatidine, the developed method was interfaced with electrospray ionization mass spectrometry. In the case of solasodine, the detection limit was estimated at 3 microg/ml for NACE-UV and at 0.05 microg/ml for NACE-MS, in the selected ion monitoring mode. PMID- 11486879 TI - Comparison of monomeric and polymeric amino acid based surfactants for chiral separations. AB - To better understand chiral recognition with polymeric amino acid based surfactants, the chromatographic performance of 18 monomeric and polymeric surfactants were compared for chiral analytes with various charge states and hydrophobicities. In this study, four amino acids (glycine, L-alanine, L-valine, and L-leucine) were chosen, and all possible combinations of the chiral single amino acid and dipeptide surfactants were synthesized. The results indicate that polymeric surfactants usually provide better chiral resolution for enantiomers of lorazepam, temazepam, 1,1'-bi-2-naphthol, and propranolol as compared to monomeric surfactants. In contrast, monomers perform better for chiral recognition of the 1,1'-bi-2-naphthyl-2,2'-diyl hydrogenphosphate enantiomers. PMID- 11486880 TI - Dialysis-solid-phase extraction combined on-line with non-aqueous capillary electrophoresis for improved detectability of tricyclic antidepressants in biological samples. AB - Dialysis-solid-phase extraction (SPE) sample pretreatment is combined on-line with non-aqueous capillary electrophoresis for the determination of tricyclic antidepressants in urine and serum. After clean-up and enrichment, the water is removed from the sample matrix and the analytes are eluted from the cartridge by means of an organic solvent. Next, the eluate is transported to the capillary and the injection is performed electrokinetically. This injection, which does not suffer from an adverse sample matrix effect because of the SPE step, results in further analyte concentration. The detection limits are in the 0.02-0.1 microg/ml range and the day-to-day repeatabilities are between 2.5 and 9.5%, which is quite satisfactory. PMID- 11486881 TI - Separation of plant membrane lipids by multiple solid-phase extraction. AB - Plant membrane lipids were separated by multiple solid-phase extraction (SPE) in a single run. Elution was performed continuously through the modulated stationary phase employing only non-aqueous solvent systems. At the different stages of the glycerolipid separation the SPE manifold combined arninopropyl, arninopropyl/silica gel and silica gel/aminopropyl weak anion exchanger columns. The glycerolipid extract of pigment-containing plant tissues was cleared from the pigments onto the aminopropyl column. The aminopropyl column with the glycerolipid extract was then connected to a silica gel column from which monogalactosyldiacylglycerol, phosphatidylethanolamine, phosphatidylglycerol and digalactosyldiacylglycerol were eluted as individual fractions. The elution was performed under polarity, pH and temperature gradient conditions. To continue the separation, the aminopropyl column was discarded and the silica gel column containing the remaining glycerolipid extract was connected to an aminopropyl anion exchanger column. Individual fractions of sulfoquinovosyldiacylglycerol, phosphatidylcholine and phosphatidylinositol were now eluted. The separation process was supported by ammonium counter ions and by the polarity gradient of the elution systems used. The membrane lipids were isolated from pigment containing (rice and maize leaves and rice leafy stems) and pigment-free (rice roots) tissues. The repeatability for a standard glycerolipid mixture was 2-6% (n=7), and for rice leaf lipid extracts, 3-7% (n=5). Glycerolipid recovery was 87 95%. PMID- 11486882 TI - Evaluation of different tubing geometries for high-speed counter-current chromatography. AB - Different tubing geometries were evaluated for use in high-speed counter-current chromatography. Standard round tubing was compared to rectangular and twisted rectangular tubing. The number of theoretical plates for a standard anthocyanin mixture from black currant was determined for each experiment. The results of twisted rectangular tubing were superior to a standard setup. PMID- 11486883 TI - Analysis of phenolic constituents of biological interest in red wines by high performance liquid chromatography. AB - We describe a reversed-phase HPLC method that uses gradient elution and diode array detection to determine four biologically active phenolic constituents of red wines: gallic acid, trans-resveratrol, quercetin and rutin. The method permits direct injection without sample pre-treatment. ODS Hypersil served as the stationary phase; the gradient was formed by acetic acid, methanol, and water. Each analysis required an equilibration period of 10 min and a run time of 50 min for completion. Previously, total phenols were analysed according to the Folin Ciocalteu method, using gallic acid as the standard, and the results are given as gallic acid equivalent. PMID- 11486884 TI - Separation of chondroitin sulfate and hyaluronic acid fragments by centrifugal precipitation chromatography. AB - Centrifugal precipitation chromatography (CPC) was applied for the first time to the separation of fragments of chondroitin sulfate (ChS) and hyaluronic acid (HA). The separation was performed using a gradient elution system between ethanol and water since solubility of these biopolymers highly depends on the concentration of ethanol in aqueous solution. ChS and HA were each eluted into several peaks through a flow-through UV detector at 275 nm, despite they have almost no absorbance at this wavelength in an aqueous solution. The separation was also confirmed by redissolving the dried fraction in water and measuring the absorbance at 210 nm. These results suggest that the CPC system can detect small precipitates of these biopolymers by light scattering at 275 nm. The separated fragments of biopolymers are not easily characterized because no suitable analytical method is available for identification of these compounds. However, the overall results demonstrate that CPC may be a useful separation of biopolymers such as glycosaminoglycans which quantitatively produce precipitates in an organic solvent mixture. PMID- 11486885 TI - Liquid crystalline polymers as stationary phases. IV. Chemical bonding and immobilization of the polymer on silica characterization by solid-state nuclear magnetic resonance spectroscopy. AB - Chemical bonding reaction and immobilization through low energy radiation (heating) have been investigated to fix a side-chain liquid crystalline polymer (SC-LCP) on silica particles in order to use the resulting modified silica in normal-phase HPLC. Highly stable chromatographic stationary phases are observed under excellent polymer solvent flow conditions (THF) for both methods and better column efficiencies are also exhibited towards PAHs' separation compared to the classical coated stationary phase. The characterization of these new stationary phases and the rationale for improved column stability have been investigated by solid state 13C and 29Si CP/MAS NMR spectroscopy. It is clearly shown that the chemical bonding is achieved by the classical hydrosilylation reaction between PHMS chains and vinyl modified silica. The bonded polymer is likely a copolymer than a homopolymer. The immobilization of the SC-LCP by heating results in the breaking of Si-O-Si bonds of the polysiloxane chain after the attack of the silica surface silanols. Applications to fullerenes and carotenes separation of these bonded stationary phases are compared to the separation power of a classical monomeric C18 stationary phase in NP-HPLC as n-hexane-toluene or methyl tertiobutyl ether-methanol mixtures. PMID- 11486886 TI - Microwave-assisted extraction of tanshinones from Salvia miltiorrhiza bunge with analysis by high-performance liquid chromatography. AB - A novel microwave-assisted extraction (MAE) method has been developed for the extraction and determination of tanshinones (tanshinone IIA, cryptotanshinone and tanshinone I) from the root of Salvia miltiorrhiza bunge with analysis by HPLC. Various experimental conditions were investigated to optimize the percentage extraction. Under appropriate MAE conditions, such as ethanol concentrations of 95% (v/v), MAE for 2 min, liquid/solid ratio of 10:1 (ml/g), the percentage extraction can reach high in a short time. The percentage extraction (tanshinone IIA: 0.29%; cryptotanshinone: 0.23%; tanshinone I: 0.11%) by MAE was the same or even higher than conventional extraction methods. MAE only needs 2 min, but extraction at room temperature, heat reflux extraction, ultrasonic extraction and Soxhlet extraction need 24 h, 45 min, 75 min and 90 min, respectively. MAE was also available in pilot plant form for larger scale extraction. PMID- 11486887 TI - Comparison of different fibers for the solid-phase microextraction of phthalate esters from water. AB - Solid-phase microextraction (SPME) coupled to gas chromatography-mass spectrometry (GC-MS) has been applied to determine six phthalate esters and one adipate ester in water. The SPME parameters were optimized for several commercially available fibers. A 65-microm polydimethylsiloxane-divinylbenzene (PDMS-DVB) was the fiber selected and was applied to analysis of water from the Ebro river and the industrial port of Tarragona. The studied compounds were found at concentrations ranging from 0.4 microg l(-1) for di-n-butyl phthalate ester (DnBP) to 3.2 microg l(-1) for bis(2-ethylhexyl) phthalate ester (DEHP). The linear range for real samples was from 0.1 to 10 microg l(-1) for most phthalates, and the limits of detection of the method were between 3 and 30 ng l( 1). Repeatability and reproducibility between days (n = 5) for 1 microg l(-1) samples were below 13 and 18%, respectively. PMID- 11486888 TI - Identification of lipid binders in paintings by gas chromatography. Influence of the pigments. AB - The influence of the presence and the type of pigments in the lipid binding media of paintings were studied by gas chromatography with flame ionization detector. The drying oils were linseed stand oil, poppy oil and sunflower oil, and the pigments studied were cadmium red, cobalt blue, tin white, lead white, chalk and plaster of Paris, commonly used in paintings. The results indicate that the stearic/palmitic ratio and the presence of pigments are quite stable during ageing. However, some differences in the oleic acid/palmitic acid ratio were found, depending on the type of pigment present in the lipid binding media. These variations are related to the drying effect of the pigments. The proposed method has been applied to the identification of drying oils in two samples from baroque paintings in the "Basilica de la Virgen de los Desamparados" of Valencia, Spain. PMID- 11486889 TI - Automated determination of fatty acid methyl ester and cis/trans methyl ester composition of fats and oils. AB - The determination of the fatty acid composition (as methyl esters, FAMEs) of fats and oils and their cis/trans (CTME) distribution requires a simple, but manual and time-consuming sample preparation. The so-called BF3 method is often the preferred procedure. Because FAME/CTME analyses are encountered very frequently in the food industry, an automated, robot-based alternative is proposed which uses the sodium methylate procedure. After sample weighing and the (manual) addition of heptane (2 min), a XYZ robotic autosampler is used for all remaining work, which includes reagent addition, agitation, sample settling and the final injection into the gas chromatograph (10 min). The performance of the sodium methylate and BF3 methods are compared by analysing some 30 oil and fat samples. The novel procedure is much faster (less than 15 min versus ca. 1 h) and manual sample handling is drastically decreased. The experimental results obtained with the two methods frequently are the same, while small differences can be explained by (known) differences of the two methods in the conversion of minor oil/fat constituents, such as free fatty acids, wax esters and sterol esters. In case of FAME analyses, a hot injection is to be preferred over a cold injection. The RSDs of the peak areas were 1.5% for the major fatty acids to 11% for peaks that were just above the noise level. The detection limit were approximately 0.03%. PMID- 11486890 TI - Cationic diazacrown ether for the separation of positional isomers by capillary electrophoresis. AB - The use of a positively charged diazacrown ether, N,N,N',N'-tetramethyl-1,4,10,13 tetraoxa-7,16-diazacyclooctadecane dichloride (1), in capillary electrophoresis is reported, for the first time. The addition of the cationic diazacrown 1 to the running buffer permitted the successful separation of positional isomers of aromatic anions such as naphthalenedisulfonate, naphthalenedicarboxylate and phthalate. At a concentration of 1 of 2 mM, all the analyte anions were completely separated in less than 5 min. The ion association constants of the solute anions with diazacrown 1 were evaluated and the findings are discussed. PMID- 11486891 TI - Laterally attached liquid crystalline polymers as stationary phases in reversed phase high-performance liquid chromatography. V. Study of retention mechanism using linear solvation energy relationships. AB - A linear solvation energy relationship model was used to characterize the retention behavior of a stationary phase based upon a nematic side-on liquid crystalline polymer (SOLCP) in reversed-phase liquid chromatography. The set of solutes was constituted of a high variety of compounds whose molecular sizes were considerably smaller than the mesogenic unit size. The results showed good statistical fits for these retention data in 65:35, 75:25 and 85:15 (v/v) methanol-water mobile phases. Both the cavity term and excess molar refraction are the most important favorable retention-governing parameters, whereas the solute hydrogen bond acceptor basicity is the most unfavorable retention parameter. Hydrophobicity and pi-pi interactions decrease strongly when the percentage of methanol increases, leading to an important retention decrease despite the fact that the hydrogen bond interaction weakens as the organic solvent is added. The shape recognition ability of this side-on liquid crystalline stationary phase on polycyclic aromatic hydrocarbon solutes is partly explained by the solutes' high polarizability due to the presence of pi electrons. However, the solute polarizability is not sufficient and a stationary phase's "structure effect" must to be taken into account for the shape discrimination observed. The strong interaction between liquid crystal molecules caused likely a adsorption retention mechanism rather than a partition mechanism. PMID- 11486892 TI - Macroporous copolymer matrix. IV. Expanded bed adsorption application. AB - Macroporous crosslinked hydroxyethyl methacrylate-ethylene dimethacrylate copolymeric beads (HEG beads) were synthesized by suspension polymerization in the presence of a pore generating agent. These beads were coupled to alpha cyclodextrin through a urethane spacer. These modified copolymer beads (affinity HEG beads) so prepared were evaluated for their suitability in expanded bed chromatography. The optimum thickness of the distributor plate for stable expanded bed for use in expanded bed adsorption (EBA) was established. The affinity-HEG beads are comparable in density to Streamline diethyl amino ethane (DEAE) and exhibit better mechanical stability at higher superficial velocity under fluidization. The affinity-BEG beads were used as affinity chromatography matrices for the purification of cyclodextrin glycosyltransferase. Feeding of 5 fold diluted fermented broth to the column containing affinity-HEG beads of settled bed height 7.5 cm (I.D. 26 mm and length 42 cm) at double bed expansion resulted in a sharp breakthrough curve of alpha-cyclodextrin glycosyltransferase (CGTase). The adsorbed enzyme was eluted from the bed in 50 mM Tris-HCl buffer containing 10 mM CaCl2 at 25 degrees C in packed bed configuration. PMID- 11486893 TI - Cooperation of solid granule and solvent as porogenic agents. Novel porogenic mode of biporous media for protein chromatography. AB - A novel porogenic mode, cooperation of solid granule and solvent, has been introduced to prepare a biporous medium for protein chromatography. The matrix, a ternary copolymer of glycidyl methacrylate, triallylisocyanurate and divinylbenzene, was produced by a simple in situ polymerization with granules of sodium sulfate and cyclohexanol and dodecanol as porogenic agents. Functionalized with diethylamine, the resin (denoted as Resin C) was used as an anion exchanger. The pore structure, specific surface area and chromatographic properties of Resin C were determined and compared with those of the resin with only the solvents as porogen (Resin A) and the resin with only the salt granules as porogen (Resin B). The results indicated that Resin C contained regions of micropores with a maximum at approximately 55 nm and regions of macropores with a distinct maximum near 340 nm, which swelled to about 1 microm in aqueous solution. Compared with Resins A and B, the biporous medium Resin C simultaneously possessed a high specific surface area of 37.2 m2/g and a low back-pressure at mobile phase flow velocity up to 720 cm/h. The result of dynamic porosity showed that mobile phase was able to convectively flow through the macropores in Resin C. The dynamic adsorption capacity of Resin C for bovine serum albumin was as high as 57.0 mg/ml column volume (95.0 mg/g wet resin), basically identical to its static capacity, while that of Resin A was only 1.95 mg/ml column volume (3.12 mg/g wet resin), about 3% that of its static capacity. In addition, the column efficiency of Resin C was comparable to that of Resin B, but much higher than that of Resin A, indicating that the mass transfer behavior of proteins in the column was greatly improved by convective flow through the macropores. PMID- 11486894 TI - Affinity screening by packed capillary high performance liquid chromatography using molecular imprinted sorbents. II. Covalent imprinted polymers. AB - This study concentrates on the production of covalent molecular imprint polymers (MIPs) as highly selective sorbents for nortriptyline (NOR), a representative tricyclic antidepressant (TCA). The functionalized template contains a polymerizable 4-vinylphenyl carbamate moiety used to bind the template molecule to the polymer matrix. Polymerization with a cross-linker followed by hydrolytic cleavage of the labile carbamate functionality leaves an MIP with selective binding sites capable of binding template through hydrogen bonding interactions. Demonstrated chromatographically through a "selection index", these MIPs showed high selectivity for the template molecule (NOR) among a library of structurally similar compounds. The recognition was found to correlate with structural similarity to the template compound. A direct comparison between covalent and non covalent molecular imprinting strategies reveals a great deal of improvement in the peak shape of the retained compound resulting from covalent imprinting (evidenced by peak asymmetry factors A.). PMID- 11486895 TI - Development of a simple gradient LC-IR interface for the detection of terpenoids from the alpha-pinene-ozone reaction. AB - The development of a simple interface between liquid chromatography and infrared spectroscopy (LC-IR) using a coaxial sprayer is described for less volatile analytes. The system consists of a transfer capillary, in which the analytes are transported from the separation column of the gradient-LC to the outlet of the sprayer. This transfer capillary is coaxially surrounded at the outlet by a stainless steel sprayer capillary, which is resistively heated and flushed with nitrogen gas. The samples are sprayed in the manner that the eluent is vaporized by the heated nitrogen when exiting the capillary, while the analytes are deposited on a moving slide made of infrared transparent material (ZnSe or CaF2). Afterwards the deposited compounds are analyzed with an infrared microscope in transmission. First results from reaction products of the gas phase reaction of alpha-pinene with ozone are presented. PMID- 11486896 TI - Management of neuronopathic Gaucher disease: a European consensus. PMID- 11486897 TI - Clinical course and biochemistry of sialuria. AB - Sialuria is a rare inborn error of metabolism in which excessive free sialic acid (N-acetylneuraminic acid, NeuAc) is synthesized. A defect in the feedback inhibition of UDP-N-acetylglucosamine (UDP-GlcNAc) 2-epimerase by the end-product of the sialic acid synthetic pathway, CMP-NeuAc, is the mechanism underlying this overproduction. Recent evidence suggests that sialuria is an autosomal dominant disorder. Only five patients have been documented to have such an enzymatic defect. We report a longitudinal study of one of the original sialuria patients, to age 11 years. Although he has coarse features and massive hepatomegaly, he has shown normal growth and relatively normal development. Pulmonary function testing showed minimal small airway obstruction. At 11 years, he developed intermittent abdominal pain and transient transaminase elevation above his baseline. Sialuria should be considered in the differential diagnosis of a patient with a phenotype suggestive of a mucopolysaccharidosis or oligosaccharidosis in the absence of developmental regression or prominent dysostosis multiplex. We recommend close monitoring of liver and pulmonary function in sialuria patients. PMID- 11486898 TI - Intermediates of unsaturated fatty acid oxidation are incorporated in triglycerides but not in phospholipids in tissues from patients with mitochondrial beta-oxidation defects. AB - The fatty acid composition was determined of liver, skeletal muscle and heart obtained post mortem from patients with medium-chain acyl-CoA dehydrogenase deficiency (MCADD), multiple acyl-CoA dehydrogenase deficiency (MADD) and very long-chain acyl-CoA dehydrogenase deficiency (VLCADD). Increased amounts of 4 decenoic acid 10:1(n-6), 5-dodecenoic acid 12:1(n-7), 5-tetradecenoic acid 14:1(n 9), 5,8-tetradecadienoic acid 14:2(n-6) and 7,10-hexadecadienoic acid 16:2(n-6)- intermediates of unsaturated fatty acid oxidation--were found. Fractionation into different lipid classes showed that these fatty acids were exclusively present in the triglyceride fraction. They could not be detected in the free fatty acid fraction or in the phospholipid fraction. Our results suggest that intermediates of unsaturated fatty acid oxidation that accumulate as a consequence of MCADD, MADD and VLCADD are transported to the endoplasmic reticulum for esterification into neutral glycerolipids. The pattern of accumulation is characteristic for each disease, which makes fatty acid analysis of total lipid of post-mortem tissues a useful tool in the detection of mitochondrial fatty acid oxidation defects in patients who died unexpected, for example with sudden infant death syndrome. PMID- 11486899 TI - A splice mutation in the GTP cyclohydrolase I gene causes dopa-responsive dystonia by exon skipping. AB - Four different mutations in the GTP cyclohydrolase I gene were found (P199L, M211V, IVS5+1G>A, G203R) in 6 out of 33 families with dopa-responsive dystonia. A splice mutation (IVS5+1G>A) located at the border of exon 5 to intron 5 was found in one of these families. Three members of the family carry the IVS5+1G>A mutation on one allele, inherited from the father to the daughter and son. Examination of the mRNA showed an exon 5 skipping that results in a reduction of enzyme activity in cultured fibroblasts to 4-17% compared to controls. The father and daughter never had clinical symptoms of dopa-responsive dystonia. The son was symptomatic at the age of 3 years and was treated successfully with L dopa/carbidopa. After 20 years this therapy was terminated and for the next 6 years he was free of symptoms. With increased motoric activity, symptoms reappeared and the therapy was reintroduced. PMID- 11486901 TI - Plasma phenylalanine is associated with decreased serum ubiquinone-10 concentrations in phenylketonuria. AB - Decreased serum ubiquinone-10 concentrations is a common condition in patients with phenylketonuria (PKU) under dietary treatment. Our aim was to investigate the implication of the metabolic abnormalities of PKU (low concentrations of tyrosine and high concentrations of phenylalanine) and the effect of treatment with phenylalanine-restricted diets in decreased ubiquinone-10 concentrations in PKU patients. We studied 30 PKU patients (age range 5 months to 35 years; median age 7 years) under dietary treatment. Correlation between plasma tyrosine or phenylalanine and serum ubiquinone-10 concentrations was investigated. Daily cholesterol intake was calculated from the data obtained through a dietary questionnaire. The index of dietary control (IDC) was calculated as the average of the medians of plasma phenylalanine concentrations obtained every 6 months in the metabolic control of patients. Negative correlations were observed between serum ubiquinone and the IDC (r=-0.46; p<0.01) in PKU patients. No correlation was observed between tyrosine or daily cholesterol intake and serum ubiquinone concentrations. After adjustment for daily cholesterol intake by multiple linear regression analysis, for each 113 units of increase in IDC values serum ubiquinone decreased 0.1 micromol/L. According to our results, the main factor associated with the decreased serum ubiquinone concentrations was high plasma phenylalanine values. Although daily cholesterol intake seems to be associated with ubiquinone concentrations, it may not be relevant if we take into account the low intake of cholesterol in treated PKU patients. PMID- 11486900 TI - Tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency in Dutch neonates. AB - Four neonates with a positive phenylalanine screening test (Phe concentrations between 258 and 1250 micromol/L) were investigated further to differentiate between phenylalanine hydroxylase (PAH) deficiency and variant hyperphenylalaninaemia (HPA) forms. In patients 1 and 2 a tetrahydrobiopterin (BH4) load caused a significant decrease of the plasma Phe levels. A combined phenylalanine/BH4 loading test was performed in patients 2, 3 and 4. In the latter two patients, plasma Phe concentrations completely normalized within 8 h after the BH4 load (20 mg/kg). Basal urinary pterins were normal in all four patients. The activity of dihydropteridine reductase (DHPR) was normal in patients 1, 2 and 3 and 50% of control values in patient 4 (not in the range of DHPR-deficient patients). In patient 3 a subsequent phenylalanine loading test with concomitant analysis of plasma biopterins revealed a normal increase of biopterin, excluding a BH4 biosynthesis defect. Pterins and neurotransmitter metabolites in CSF of patients 1, 3 and 4 were normal. DNA mutations detected in the PAH gene of patients 1-4 were A313T, and L367fsinsC; V190A and R243X; A300S and A403V; R241C and A403V. The results are suggestive for mutant PAH enzymes with decreased affinity for the cofactor BH4. PMID- 11486902 TI - Factor V Leiden mutation in Turkish patients with homozygous cystathionine beta synthase deficiency. AB - Venous and arterial thromboembolism can occur in patients with homocystinuria. Resistance to activated protein C, which is caused by a single point mutation in the gene for factor V, renders an individual at risk for thrombosis. It has been suggested that coexistence of hereditary homocystinuria and factor V Leiden mutation might jointly play a role in the development of thrombosis. We analysed six patients with homocystinuria due to cystathionine beta-synthase deficiency for factor V Leiden and prothrombin G20210A mutations. Only one patient was found to have the factor V Leiden mutation in homozygous form and this patient had suffered from severe thrombosis. One patient was found to be heterozygous with no documented thrombosis. None of the patients had prothrombin G20210A mutation. We stress the necessity for screening for known thrombophilic risk factors in patients with cystathonine beta-synthase deficiency. The coexistence of the factor V Leiden mutation can cause severe thrombotic events in patients with homocystinuria. PMID- 11486903 TI - Detection of neonatal argininosuccinate lyase deficiency by serum tandem mass spectrometry. AB - Argininosuccinate lyase (ASL) deficiency (McKusick 207900) is an inborn error of the urea cycle. The leading symptom is progressive hyperammonaemia, which is a life-threatening condition, particularly in patients with a neonatal onset. Early diagnosis and treatment of the hyperammonaemia are necessary to improve survival and the long-term outcome of ASL-deficient patients. Currently, the diagnosis of ASL deficiency is based on the measurement of urea cycle intermediates and amino acids by automated quantitative ion exchange chromatography in plasma and urine. Here, we report a newborn presenting with coma and severe hyperammonaemia. ASL deficiency was suspected on the basis of an adapted tandem mass spectrometric (MS MS) procedure which allows determination of argininosuccinate in addition to the amino acids in serum samples. MS-MS measurements revealed a characteristic increase of argininosuccinate, a moderate increase of citrulline, and lowered levels of arginine and ornithine in the serum of the patient. The diagnosis was confirmed by the detection of a novel homozygous frameshift mutation in exon 14 of the argininosuccinate lyase gene. We propose MS-MS as a diagnostic tool suitable for the rapid detection of specific alterations in the amino acid spectra caused by ASL deficiency. PMID- 11486904 TI - Frontal lobe dementia with abnormal cholesterol metabolism and heterozygous mutation in sterol 27-hydroxylase gene (CYP27). AB - Of the primary dementing disorders that cause frontotemporal dementia, the best known is Pick disease. We report on a 44-year-old woman with progressive frontal lobe dementia and spastic paraplegia. Examination revealed increased serum levels of cholestanol with abnormal cholesterol metabolism and a heterozygous mutation of the sterol 27-hydroxylase gene (CYP27). Biochemical findings were compatible with cerebrotendinous xanthomatosis (CTX); however, the clinical manifestations were very dissimilar. To our knowledge, a symptomatic carrier of this mutation among CTX patients has not been reported. We speculate that the present patient has a previously undescribed neurodegenerative disease related to abnormal cholesterol metabolism with this heterozygous mutation. PMID- 11486905 TI - Carrier detection and rapid newborn diagnostic test for the common Y393N maple syrup urine disease allele by PCR-RFLP: culturally permissible testing in the Mennonite community. AB - The turnaround time for diagnosis of maple syrup urine disease (MSUD) by classic serum amino acid analyses often requires 3-4 days. This is due to the need for branched-chain amino acids (BCAA) to accumulate in the serum of the newborn before testing. The accumulation of BCAAs in infants with MSUD during this time increases the risk of the infant becoming clinically symptomatic. We have developed a noninvasive DNA-based mismatch PCR-RFLP assay for the Y393N BCKDHA allele (E1alpha gene of the branched chain alpha-keto acid dehydrogenase complex), the primary cause of MSUD in Old Order Mennonite communities. The homozygosity and high frequency of this mutation in the Mennonite community and its prevalence in compound heterozygote non-Mennonite MSUD patients is of significance. We describe carrier testing, present the results of nine newborns diagnostically evaluated for the Y393N BCKDHA allele, and demonstrate the efficacy of this PCR-RFLP assay for determining clinical status within 24 h after birth. Analyses within the first 24 h of life allow for immediate diagnosis and treatment of infants homozygous for the Y393N MSUD defect. This is a significant improvement over the time required by current serum amino acid analysis methods. PMID- 11486906 TI - Massive 5-oxoprolinuria with normal 5-oxoprolinase and glutathione synthetase activities. PMID- 11486907 TI - Sudden death in a Korean infant with very long-chain acyl-CoA dehydrogenase deficiency. PMID- 11486908 TI - Ketoacidosis: an unusual presentation of MELAS. PMID- 11486909 TI - Mevalonate kinase deficiency in a child with periodic fever and without hyperimmunoglobulinaemia D. PMID- 11486910 TI - Mevalonic aciduria and hyper-IgD syndrome: two sides of the same coin? PMID- 11486911 TI - Gabapentin interference with urine histidine as measured by the Beckman amino acid analyser. PMID- 11486912 TI - Compound heterozygosity in four asymptomatic siblings with medium-chain acyl-CoA dehydrogenase deficiency. PMID- 11486913 TI - Splenectomy in two siblings with G-CSF-dependent glycogen storage disease type Ib. PMID- 11486914 TI - Carnitine-responsive carnitine insufficiency in a case of mtDNA 8993T>C mutation associated Leigh syndrome. PMID- 11486915 TI - Sustained oral lysine supplementation in ornithine delta-aminotransferase deficiency. AB - Oral lysine administration to three patients with B6-nonresponsive gyrate atrophy reduced plasma ornithine concentrations by 21-31% within 1-2 days. No further reduction was noted with time. PMID- 11486916 TI - Overconfidence effects in category learning: a comparison of connectionist and exemplar memory models. AB - Exemplar and connectionist models were compared on their ability to predict overconfidence effects in category learning data. In the standard task, participants learned to classify hypothetical patients with particular symptom patterns into disease categories and reported confidence judgments in the form of probabilities. The connectionist model asserts that classifications and confidence are based on the strength of learned associations between symptoms and diseases. The exemplar retrieval model (ERM) proposes that people learn by storing examples and that their judgments are often based on the first example they happen to retrieve. Experiments 1 and 2 established that overconfidence increases when the classification step of the process is bypassed. Experiments 2 and 3 showed that a direct instruction to retrieve many exemplars reduces overconfidence. Only the ERM predicted the major qualitative phenomena exhibited in these experiments. PMID- 11486917 TI - Exemplar accounts of blending and distinctiveness effects in perceptual old-new recognition. AB - Recent findings from the perceptual old-new recognition literature indicate that observers have extremely high false-alarm rates to new items that are "blends" of old ones. In addition, evidence suggests that "distinctive" old items--that is, those located in isolated regions of the similarity space--are recognized with higher probability than are typical old items. Both types of phenomena challenge the predictions of global-familiarity exemplar models of perceptual old-new recognition, which posit that the probability that an observer judges an item as old is based on its summed similarity to previously presented exemplars. In the present research the authors pursued these blending and distinctiveness effects by testing paradigms in which similarity relations among objects are highly controlled and in which the variables of blending and distinctiveness are not confounded with other properties associated with the individual objects themselves. In contrast to previous results, the authors found effects of blending and distinctiveness that are compatible with the predictions of a pure summed-similarity exemplar model. PMID- 11486918 TI - The effect of shared structure and content on reading nonwords: evidence for a CV skeleton. AB - Four experiments examined the effect of shared skeletal structure versus content overlap on naming printed nonwords. Experiments 1-2 compared priming among nonwords sharing either skeletal structure and content (e.g., dus-DUS) or structure alone (e.g., pid-BAF) with controls that differed from the target in the number of skeleton slots (e.g., pid-BAF vs. plid-BAF). Conversely, in Experiments 3-4, same-versus different-structure primes contrasted only in the ordering of CV skeletal slots (e.g., fap-DUS vs. ift-DUS). Priming effects were modulated by shared content and skeletal similarity. The sensitivity of skeletal priming to the abstract arrangement of consonants and vowels suggests that skeletal representations assign distinct slots for consonants and vowels. Readers' sensitivity to skeletal structure in nonword identification indicates that assembled phonological representations are constrained by linguistic knowledge. PMID- 11486919 TI - Semantic and phonological activation in noun and pronoun production. AB - Speakers can refer to objects and other entities by nouns or pronouns. The present article investigated the production of gender-marked pronouns in German. Four picture-word interference experiments are reported, addressing 2 questions. First, is the lemma of a referent noun (i.e., the representation of the referent noun's semantic and syntactic properties) accessed when producing a pronoun? Second, if so, is this access confined to the lemma, or will the referent noun's phonological form be activated, too? The results suggest that in generating pronouns, speakers accessed the lemma of the referent noun, whereas its phonological form was not substantially activated. The results are discussed in the context of other recent experimental studies of pronoun and noun production. PMID- 11486920 TI - Rowed to recovery: the use of phonological and orthographic information in reading Chinese and English. AB - To examine how readers of Chinese and English take advantage of orthographic and phonological features in reading, the authors investigated the effects of spelling errors on reading text in Chinese and English using the error disruption paradigm of M. Daneman and E. Reingold (1993). Skilled readers in China and the United States read passages in their native language that contained occasional spelling errors. Results showed that under some circumstances very early phonological activation can be identified in English, but no evidence for early phonology was found in Chinese. In both languages, homophone errors showed a benefit in measures of later processing, suggesting that phonology helps readers recover from the disruptive effects of errors. These results suggest that skilled readers take advantage of the special features of particular orthographies but that these orthographic effects may be most pronounced in the early stages of lexical access. PMID- 11486921 TI - Visual short-term memory is influenced by haptic perception. AB - The present study investigated whether and how visual memory and haptic perception are related. Participants were required to compare a visual reference velocity with a visual test velocity separated by a 4-s interval. During the retention interval, a fast or slow hand movement was performed. Although the hand movement was not visible, effects of the speed of the distracting body movement occurred. Slow movements resulted in a lowering of the represented visual velocity, whereas fast movements heightened the represented velocity. Subsequent experiments extended the effect to body movements that differed from the visual motion and ruled out the possibility that the effect was due to changes in visual perception or interference from semantic, verbal, and acoustic memory codes. Perhaps haptic velocity information and visual velocity information stored in short-term memory are blended. PMID- 11486923 TI - Metacognition in motor learning. AB - Research on judgments of verbal learning has demonstrated that participants' judgments are unreliable and often overconfident. The authors studied judgments of perceptual-motor learning. Participants learned 3 keystroke patterns on the number pad of a computer, each requiring that a different sequence of keys be struck in a different total movement time. Practice trials on each pattern were either blocked or randomly interleaved with trials on the other patterns, and each participant was asked, periodically, to predict his or her performance on a 24-hr test. Consistent with earlier findings, blocked practice enhanced acquisition but harmed retention. Participants, though, predicted better performance given blocked practice. These results augment research on judgments of verbal learning and suggest that humans, at their peril, interpret current ease of access to a perceptual-motor skill as a valid index of learning. PMID- 11486922 TI - Source ROCs are (typically) curvilinear: comment on Yonelinas (1999). AB - On the basis of his assumption that recollection is a threshold process, A. P. Yonelinas (1999) predicted linear source-identification receiver operating characteristics (ROCs) and recently reported data that were consistent with this prediction. In this article, the authors present data showing curvilinear source identification ROCs across various encoding and test conditions. On the basis of the source-monitoring framework (e.g., M. K. Johnson, S. Hashtroudi, & D. S. Lindsay, 1993), the authors argue that curvilinearity of source-identification ROCs is a result of differences in the qualitative characteristics of memories rather than simply the influence of undifferentiated familiarity as the dual process model might suggest. PMID- 11486924 TI - Presentation modality and mode of recall in verbal false memory. AB - False memories were investigated for aurally and visually presented lists of semantically associated words. In Experiment 1, false written recall of critical intrusions was reliably lower following visual presentation compared with aural presentation. This presentation modality effect was attributed to the use of orthographic features during written recall to edit critical intrusions from visually presented lists. As predicted by this hypothesis, the modality effect was eliminated when the mode of recall was spoken rather than written. In Experiment 2, the modality effect in written recall was again replicated and then eliminated with an orienting task that ensured orthographic encoding even of aurally presented words. Thus, the modality effect appears to depend on using orthographic information to distinguish true from false verbal memories. PMID- 11486925 TI - Comparing recollective experience in true and false autobiographical memories. AB - This study investigated whether true autobiographical memories are qualitatively distinct from false autobiographical memories using a variation of the interview method originally reported by E. F. Loftus and J. Pickrell (1995). Participants recalled events provided by parents on 3 separate occasions and were asked to imagine true and false unremembered events. True memories were rated by both participants and observers as more rich in recollective experience and were rated by participants as more important, more emotionally intense, as having clearer imagery, and as less typical than false memories. Rehearsal frequency was used as a covariate, eliminating these effects. Imagery in true memories was most often viewed from the field perspective, whereas imagery in false memories was most often viewed from the observer perspective. More information was communicated in true memories, and true memories contained more information concerning the consequences of described events. Results suggest repeated remembering can make false memories more rich in recollective experience and more like true memories. Differences between true and false memories suggest some potentially distinct characteristics of false memories and provide insight into the process of false memory creation. PMID- 11486926 TI - A demonstration and comparison of two types of inference-based memory errors. AB - Participants viewed slides depicting ordinary routines (e.g., going grocery shopping) and later received a recognition test. In Experiment 1, there was higher recognition confidence to high-schema-relevant than to low-schema-relevant items. In Experiment 2, participants viewed slide sequences that sometimes contained a cause (e.g., woman taking orange from bottom of pile) but not an effect scene (oranges on floor), or an effect but not a cause scene. Participants mistook new cause scenes as old when they viewed the effect; false alarms to cause scenes and high-schema-relevant items increased with retention interval. Experiment 3 showed that the backward inference effect was accompanied by false explicit recollection, whereas false alarms to schema-high foils were based on familiarity. This suggests that the 2 types of inferential errors are produced by different underlying mechanisms. PMID- 11486927 TI - On the dual effects of repetition on false recognition. AB - The effects of study-list repetition on false recognition of semantic associates were examined using aging (Experiment 1) and recognition time pressure (Experiment 2). Participants studied word lists, each of which was composed of high associates to a single, unstudied word (the critical lure). Under normal testing circumstances, young adult participants (ages 19-26) falsely endorsed fewer critical lures associated with lists that had been presented multiple times than lists presented only once. However, young participants tested under time pressure and older participants (ages 67-85) endorsed a greater number of critical items associated with lists presented thrice than with lists presented once. The results suggest dual bases for the recognition decision, one of which is based on the rapid spread of activation within domains of semantic similarity and the other of which functions to attribute that activation to likely sources and set appropriate decision criteria. The latter capacity is compromised both under conditions of time pressure and in the elderly. PMID- 11486928 TI - Removing irrelevant information from working memory: a cognitive aging study with the modified Sternberg task. AB - Two experiments with a modified Sternberg recognition task explored the ability of young and old adults to remove irrelevant information from working memory. The task involved 2 memory sets, 1 of which was later cued as irrelevant. The recognition probe was presented at a variable time after the cue. Two indicators of inhibition, the setsize effect of the irrelevant set and the reaction time cost of intrusion probes (i.e., negative probes present in the irrelevant list), were dissociated. Irrelevant setsize effects lasted less than 1 s after the cue and did not differ between old and young adults. Intrusion costs lasted up to 5 s and were disproportionally large for old adults. With the additional requirement to remember both lists until after the probe, young adults' intrusion costs in Experiment 2 were equivalent to those of old adults in Experiment 1, but the setsize effects of the irrelevant set was larger. The results are compatible with a dual-process model of recognition in combination with a working-memory model distinguishing the focus of attention from the activated portion of long-term memory. PMID- 11486929 TI - Measuring automatic retrieval. AB - A variety of procedures have been used to assess automatic retrieval effects on memory, including implicit memory tests and the process dissociation approach. Theoretical concerns with each are summarized prior to describing a procedure for evaluating automatic retrieval that is based on retrieval speed. Specifically, in a speeded implicit task, participants were encouraged to complete word stems using strictly automatic retrieval by presenting several practice test trials that did not allow responding based on previously studied items and by encouraging speed of responding. This speeded implicit task was compared with a condition in which conscious retrieval of studied information was not possible and a condition in which conscious retrieval was required, providing converging evidence to support the hypothesis that the speeded implicit procedure can yield pure estimates of automatic retrieval. Furthermore, evidence from a standard implicit memory task yielded comparable data that suggests that participants engaged automatic retrieval processes on this task also. PMID- 11486930 TI - Implicit learning of sequences of tasks. AB - Task sets can be configured in advance of performing a new task. However, the degree to which advance information is actually used for advance configuration depends on the nature of the available information. The role of implicit learning was explored in 2 experiments by means of a modified serial reaction time task with repeated sequences of 4 dimensionally organized tasks. Although there was clear evidence for implicit learning of the sequence (of length 8), the learning was not associated with a reduction of shift costs, either with a short (200 ms) or with a long (1,200 ms) response-stimulus interval. In contrast, a reduction of shift costs was observed when external precues were introduced in a 3rd experiment. According to these results, the sequences of stimulus features that serve as cues for the tasks to perform on the stimuli are learned, but the representation of the features is void of their task-associated meanings. PMID- 11486931 TI - Journey to the center of the category: the dissociation in amnesia between categorization and recognition. AB - The authors' theoretical analysis of the dissociation in amnesia between categorization and recognition suggests these conclusions: (a) Comparing to-be categorized items to a category center or prototype produces strong prototype advantages and steep typicality gradients, whereas comparing to-be-categorized items to the training exemplars that surround the prototype produces weak prototype advantages and flat typicality gradients; (b) participants often show the former pattern, suggesting their use of prototypes; (c) exemplar models account poorly for these categorization data, but prototype models account well for them; and (d) the recognition data suggest that controls use a single comparison exemplar-memorization process more powerfully than amnesics. By pairing categorization based in prototypes with recognition based in exemplar memorization, the authors support and extend other recent accounts of cognitive performance that intermix prototypes and exemplars, and the authors reinforce traditional interpretations of the categorization-recognition dissociation in amnesia. PMID- 11486932 TI - Arranging conditions for learning. PMID- 11486933 TI - U.S. health care system. PMID- 11486934 TI - U.S. health care system. PMID- 11486935 TI - Other perspectives. PMID- 11486936 TI - Aphakic hydrogel contact lens fitting on a monocular canine: a case report. AB - BACKGROUND: Though plano bandage contact lenses used for therapeutic purposes are not uncommon for dogs, no literature regarding contact lenses to correct aphakic canines currently exists. CASE REPORT: Oliver, a 7-year-old terrier mix, was aphakic in his left eye and essentially blind in his right eye as the result of a large retinal detachment. Surgical complications and endothelium damage contraindicated an intraocular lens implant in his left eye. While co-managed with his veterinary ophthalmologist, Oliver was prescribed an aphakic hydrogel contact lens to improve his monocular vision. CONCLUSION: Oliver was successful (by clinical criteria) with his contact lens. Although it is difficult to quantify his vision without a visual evoked potential, Oliver appeared more attentive and confident with the contact lens. This case report demonstrates a successful canine aphakic contact lens fit through the efforts of his owners, co managing veterinary ophthalmologist, and optometrist. PMID- 11486937 TI - Eye conditions among 5- to 7-year-old Asian-Pacific Islander schoolchildren in Southern California. AB - BACKGROUND: The prevalence rates and specific risk factors for development of eye conditions among Asian-Pacific Islander schoolchildren are unknown. This information is necessary to understand their eye care needs and to improve access to eye care for this rapidly growing population. METHODS: Data obtained from a computer database of vision screening examinations conducted by the UCLA Mobile Eye Clinic from October 1987 through December 1996 were analyzed for 2,687 Asian Pacific Islander schoolchildren between 5 and 7 years of age. RESULTS: Myopia (8.9%) and astigmatism (15.8%) represented the majority of visual disorders. Mean myopic refractive error was -1.21 +/- 0.83 D among bilateral myopes. Color-vision deficiency was prevalent among 2.8% of male children, extraocular muscle imbalance among 3.0% of children, and amblyopia among 1.0% of children. All other conditions were present in less than 4% of children screened. CONCLUSIONS: Asian Pacific Islander schoolchildren may be at higher risk for development of juvenile myopia than white, Hispanic, and African-American children. Prevalence rates of other disorders are comparable to those obtained for other ethnic groups. This information can be used in planning to meet the eye care needs of Asian-Pacific Islander schoolchildren. PMID- 11486938 TI - Sorsby's fundus dystrophy. AB - BACKGROUND: Sorsby's fundus dystrophy describes the condition in which an autosomal-dominant inherited macular dystrophy with bilateral central visual loss is accompanied by progressive atrophy of the peripheral choroid and, subsequently, the outer retina. The first symptom of this condition is nyctalopia, whereas (typically) the first sign is the appearance of white-yellow deposits in the posterior pole. CASE REPORT: We report a case of Sorsby's fundus dystrophy, manifested with central visual loss in the fifth decade of life. A battery of tests--including ophthalmoscopy, electroretinography, color vision, Pelli-Robson contrast sensitivity, and fluorescein angiography--were used to evaluate the condition. CONCLUSIONS: While advances in understanding the etiology of Sorsby's fundus dystrophy have been made, the condition remains difficult to treat and the outcome is often devastating. The benefits of various methods of treatment, such as laser photocoagulation and dietary supplementation are unclear Patients affected by this condition should be managed in a manner similar to that for end-stage sufferers of age-related macular degeneration. PMID- 11486939 TI - A comparison of the visual symptoms between ADD/ADHD and normal children. AB - BACKGROUND: Attention Deficit Disorder (ADD) and Attention Deficit Hyperactivity Disorder (ADHD) are commonly diagnosed conditions that affect the lives of many individuals Diagnosis of ADD/ADHD is based primarily on observation, although more-objective tests are being developed. Ritalin is the most-popular treatment for ADD/ADHD. Literature indicates the ADD/ADHD patient may experience visual system dysfunctions. The purpose of this article is to document visual system symptoms that may coexist with treated ADD/ADHD. METHODS: Forty-three subjects were separated into two groups, which were matched for age and gender The experimental group had previously been diagnosed as ADD/ADHD and was under pharmacological treatment. The control group was comprised of non-ADD/ADHD children. A modified College of Optometrists in Vision Development (COVD) Quality of Life Outcomes Assessment was given to the parents of each child, which the child and parent completed together. The results between the experimental and control group were then compared. RESULTS: Results show that ADD/ADHD subjects report and/or experience more symptoms of visual system dysfunction than age matched norms. Fourteen of the 33 symptoms were found to be significantly more severe in the ADD/ADHD group than in the control group. These symptoms were relatively evenly divided between four major symptom groups. CONCLUSIONS: ADD/ADHD children, even with current medical treatment, exhibit more visual and quality of life symptoms than do a similar group of non-ADD/ADHD children. PMID- 11486941 TI - What exit strategy is right for you? PMID- 11486940 TI - Microcysts: clinical significance and differential diagnosis. AB - BACKGROUND: Microcysts are the most-distinctive and easily detectable indicator of contact lens-induced hypoxia. They should not be confused with cyst-like inclusions that occur in conditions such as Meesmann's dystrophy, bullous keratopathy, and Cogan's microcystic dystrophy, or with mucin balls, vacuoles, microcystic edema, and infiltrates. METHODS: Data from published literature and recent data from The Cornea and Contact Lens Research Unit (CCLRU) clinical trials involving low Dk and prototype high Dk soft contact lenses (SCLs) were examined. RESULTS: Extended wear with low Dk SCLs induces significant numbers of microcysts, whereas extended wear with high Dk SCLs does not. Subjects who transfer from low Dk to high Dk lenses have an initial increase in the number of microcysts after seven days that declines to normal levels over 1 to 3 months. Microcysts can be differentiated from other ocular conditions as they show reversed illumination, are 10- to 50-microm irregularly shaped dots, and are often associated with lens-induced hypoxia. CONCLUSIONS: Microcysts are easily observed and differentiated from other conditions in clinical practice. On average, fewer than ten microcysts per eye occur in nonhypoxic lens wear and, on average, more than 20 microcysts per eye are an indication of chronic hypoxia. The increase in microcyst numbers after transfer from low to high Dk lens wear is transitory and does not necessitate a period of no lens wear. PMID- 11486943 TI - Driving the sunglass market with technology. PMID- 11486942 TI - Good patient records: your keys to successful practice. PMID- 11486944 TI - Why optometrists need to market. PMID- 11486945 TI - Get on board with AOA Back-in-School Program. PMID- 11486946 TI - New avenues in dinitrogen fixation research. AB - The present article focuses on a recent finding concerning dinitrogen fixation by using titanium oxide/conducting polymer composite systems and its comparison with an earlier fixation method (Schrauzer process) that makes use of a powdered titanium oxide. Both processes work under the stimulus of light at room temperature and pressure, but dinitrogen is fixed to a solid ammonium salt crystal in the former and to a gaseous ammonia molecule in the latter process. Differences in the physicochemical concepts between the two processes are discussed. PMID- 11486947 TI - Point-dipole approximation of the exciton coupling model versus type of bonding and of excitons in porphyrin supramolecular structures. AB - The application of the exciton coupling model to interacting porphyrin chromophores is discussed. Covalently bonded systems and ionic or electrostatically bonded homoassociates require different orientations of the transition dipole moments in order to explain the experimental results: according to the symmetry of the assembly for covalently bonded porphyrins, and assuming isolated chromophores for ionic bonded porphyrins. Further, for covalently bonded systems, an extended exciton coupling has been demonstrated, but the ionic systems are in agreement with non-extended couplings. The relation of these facts to a molecular description of solid-state Wannier-Mott or Frenkel excitons is briefly discussed. PMID- 11486948 TI - The rearrangement of the cubane radical cation in solution. AB - The rearrangement of the cubane radical cation (1*+) was examined both experimentally (anodic as well as (photo)chemical oxidation of cubane 1 in acetonitrile) and computationally at coupled cluster, DFT, and MP2 [BCCD(T)/cc pVDZ//B3LYP/6-31G* ZPVE as well as BCCD(T)/cc-pVDZ//MP2/6-31G* + ZPVE] levels of theory. The interconversion of the twelve C2v degenerate structures of 1*+ is associated with a sizable activation energy of 1.6 kcalmol(-1). The barriers for the isomerization of 1*- to the cuneane radical cation (2*+) and for the C-C bond fragmentation to the secocubane-4,7-diyl radical cation (10*+) are virtually identical (deltaH0++ = 7.8 and 7.9 kcalmol(-1), respectively). The low-barrier rearrangement of 10*+ to the more stable syn-tricyclooctadiene radical cation 3*+ favors the fragmentation pathway that terminates with the cyclooctatetraene radical cation 6*+. Experimental single-electron transfer (SET) oxidation of cubane in acetonitrile with photoexcited 1,2,4,5-tetracyanobenzene, in combination with back electron transfer to the transient radical cation, also shows that 1*+ preferentially follows a multistep rearrangement to 6*+ through 10*+ and 3*+ rather than through 2*+. This was confirmed by the oxidation of syn tricyclooctadiene (3), which, like 1, also forms 6 in the SET oxidation/rearrangement/electron-recapture process. In contrast, cuneane (2) is oxidized exclusively to semibullvalene (9) under analogous conditions. The rearrangement of 1*+ to 6*+ via 3*+, which was recently observed spectroscopically upon ionization in a hydrocarbon glass matrix, is also favored in solution. PMID- 11486949 TI - Electrochromics by intramolecular redox switching of single bonds. AB - Two moieties of mono- and trimethincyanines as well as those of styryl dyes were connected by a saturated alkyl tether made from compounds 3a-c, 5, 7, and 9a,b. In most cases, cyclic voltammetry and spectroelectrochemistry for these dyes together with the data for their monomeric models 4, 6, 8, and 10 reveal electrochemically irreversible transfer of two electrons but chemically reversible reaction and discoloration both on reduction and oxidation. Discoloration is interpreted as intramolecular formation of a single bond, which on redox breaking regenerates the starting colored species. Therefore, the investigated dyes exemplify a new general principle for electrochromics. PMID- 11486950 TI - A liquid-crystalline silsesquioxane dendrimer exhibiting chiral nematic and columnar mesophases. AB - A hexadecamer, first-generation, octasilsesquioxane liquid-crystalline dendrimer was synthesized by a platinum-catalyzed hydrosilylation reaction of the parent first-generation vinyl octasilsesquioxane dendrimer with a modified, laterally substituted mesogen. The structure and purity of the octasilsesquioxane substrate was confirmed by 1H, 13C, and 29 Si NMR spectroscopy, microanalysis, and size exclusion chromatography (SEC). The mesogenic substrate was found to exhibit only a chiral nematic phase, whereas the resulting hexadecamer dendrimer displays enantiotropic chiral nematic, disordered hexagonal columnar, and disordered rectangular columnar phases, with a glass transition below room temperature. The lateral or side-on attachment of the mesogen to the dendritic core was found to be a key design feature in the formation of the mesophases. PMID- 11486951 TI - Ligand-to-metal ratio controlled assembly of tetra- and hexanuclear clusters towards single-molecule magnets. AB - A simple template-mediated route, starting from triethalolamine 1, sodium hydride or caesium carbonate, and iron(III) chloride led to the six- and eight-membered iron coronates [Na c [Fe6[N(CH2CH2O)3]6]]+ (2) and [Cs c (Fe8[N(CH2CH2O)3]8]]+ (3). In the reaction of N-methyldiethanolamine 4 (H2L1) or N-(2,5 dimethylbenzyl)iminodiethanol 6 (H2L2) with calcium hydride followed by addition of a solution of iron(III) chloride, the neutral unoccupied coronands [Fe6Cl6(L1)6] (5) and [Fe6Cl6(L2)6] (7) were formed. Subsequent exchange of the chloride ions of 7 by bromide or thiocyanate ions afforded the ferric wheels [Fe6Br6(L2)6] (8) or [Fe6(NCS)6(L2)6] (9), respectively. Titration experiments of solutions of dianion (L1)2- with iron(III) chloride in THF revealed interesting mechanistic details about the self-assembling process leading to 5. At an iron/ligand ratio of 1:1.5 star-shaped tetranuclear [Fe[Fe(L1)2]3] (11) was isolated. However, at an iron/ligand ratio of 1:2, complex 11 was transformed into the ferric wheel 5. It was shown, that the interconversion of 5 and 11 is reversible. Based on the mechanistic studies, a procedure was developed which works for both the synthesis of homonuclear 11 and the star-shaped heteronuclear clusters [Cr[Fe(L1)2]3] (12) and [Al[Fe(L1)2]3] (13). The structures of all new compounds were determined unequivocally by single-crystal X-ray analyses. PMID- 11486952 TI - Self-assembly and crystal structure of a novel octadecametallic square box composed of 52 single components. AB - Reaction of divalent hexacoordinate metal ions with either tetradentate N-(2 piridylmethyl)iminodiethanol (3; H2L4) or tridentate N-(2 chlorobenzyl)iminodiethanol (5; H2L5) resulted in the formation of two completely different products. Starting from nickel(II) acetate and tetradentate 3, linear trinuclear complex [[Ni(HL4)]2-Ni(OAc)2(HCO2)2] (4) was formed. However, when tridentate 5 was allowed to react with cobalt(II) acetate, the novel octadecametallic square box [Na2-(Co4(HL5)3(OAc)5]4(HCO2)2] (6) was isolated. Chiral 6 has D2-molecule symmetry and crystalizes with both enantiomers in the unit cell. The structures of all new compounds were determined unequivocally by single-crystal X-ray analyses. PMID- 11486953 TI - DFT study of the active intermediate in the Fenton reaction. AB - Density functional theory has been used to investigate the nature of the oxidizing agent in the Fenton reaction. Starting from the primary intermediate [FeII(H2O)5H2O2]2+, we show that the oxygen-oxygen bond breaking mechanism has a small activation energy and could therefore demonstrate the catalytic effect of the metal complex. The O-O bond cleavage of the coordinated H2O2, however, does not lead to a free hydroxyl radical. Instead, the leaving hydroxyl radical abstracts a hydrogen from an adjacent coordinated water leading to the formation of a second Fe-OH bond and of a water molecule. Along this reaction path the primary intermediate transforms into the [FeIV(H2O)4(OH)2]2+ complex and in a second step into a more stable high valent ferryl-oxo complex [FeIV(H2O)5O]2+. We show that the energy profile along the reaction path is strongly affected by the presence of an extra water molecule located near the iron complex. The alternative intermediate [FeII(H2O)4(OOH-)(H3O+)]2+ suggested in the literature has been also investigated, but it is found to be unstable against the primary intermediate. Our results support a picture in which an FeIV-oxo complex is the most likely candidate as the active intermediate in the Fenton reaction, as indeed first proposed by Bray and Gorin already in 1932. PMID- 11486954 TI - Allylindation of cyclopropenes in organic and aqueous media: switching the regio- and stereoselectivity based on the chelation with a hydroxyl group and the crystal structure of the cyclopropylindium product. AB - Hydroxy-bearing cyclopropenes react with allylindium reagents to undergo clean allylindation both in organic and aqueous media, in which the chelation of the hydroxyl group to indium plays the central role. The regio- and stereoselectivity have been regulated both by the location of the hydroxyl group in the molecules and the reaction solvents. In particular, the allylindation in water shows marked differences from that in organic solvents; the regio- and stereoselectivity have totally been reversed compared with those in organic solvents. Unusually stable cyclopropyl-indium compounds have been isolated from the reaction of 1-(omega hydroxyalkyl)cyclopropenes and the structure has fully been established by X-ray crystallography. PMID- 11486955 TI - Orthonitridoborate ions [BN3]6- in oxonitridosilicate cages: synthesis, crystal structure, and magnetic properties of Ba4Pr7[Si12N23O][BN3], Ba4Nd7[Si12N23O][BN3], and Ba4Sm7[Si12N23O][BN3]. AB - The isotypic title compounds Ba4Pr7[Si12N23O][BN3], Ba4Nd7[Si12N23O][BN3], and Ba4Sm7[Si12N23O][BN3] were prepared by reaction of Pr, Nd, or Sm, with barium, BaCO3, Si(NH)2, and poly(boron amide imide) in nitrogen atmosphere in tungsten crucibles using a radiofrequency furnace at temperatures up to 1650 C. They were obtained as main products (approximately 70%) embedded in a very hard glass matrix in the form of intense dark green (Pr), orange-brown (Sm), or dark red (Nd) large single crystals, respectively. The stoichiometric composition of Ba4Sm7[Si12N23O][BN3] was verified by a quantitative elemental analysis. According to the single-crystal X-ray structure determinations (Ba4Ln7[Si12N23][BN3], Z= , P6 with Ln = Pr: a = 1225.7(1), c = 544.83(9) pm, R1 = 0.013, wR2 = 0.030; Ln = Nd: a = 1222.6(1), c = 544.6(1) pm, R1 = 0.017, wR2 = .039; Ln = Sm: a = 1215.97(5), c = 542.80(5) pm, R1 = 0.047, wR2 = 0.099) all three compounds are built up by a framework structure [Si12N23O]23- of corner sharing SiX4 tetrahedrons (X = O, N). The oxygen atoms are randomly distributed over the X positions. The trigonal-planar orthonitridoborate ions [BN3]6- and also the Ln(3)3+ are situated in hexagonal cages of the framework (bond lengths Si-(N/O) 169-179 pm for Ln=Pr). The remaining Ba2+ and Ln3- ions are positioned in channels of the large-pored network. The trigonal-planar [BN3]6- ions have a B N distance of 147.1(6) pm (for Ln = Pr). Temperature-dependent susceptibility measurements for Ba4Nd7[Si12N23O][BN3] revealed Curie-Weiss behavior above 60 K with an experimental magnetic moment of muexp = 3.36(5) microB/Nd. The deviation from Curie-Weiss behavior below 60 K may be attributed to crystal field splitting of the J = 9/2 ground state of the Nd3+ ions. No magnetic ordering is evident down to 4.2 K. PMID- 11486956 TI - Helical molecular programming: folding of oligopyridine-dicarboxamides into molecular single helices. AB - Molecular strands composed of alternating 2,6-diaminopyridine and 2,6 pyridinedicarbonyl units have been designed to self-organize into single stranded helical structures upon forming intramolecular hydrogen bonds. Pentameric strands 11, 12, and 14, heptameric strands 1 and 20, and undecameric strand 15 have been synthesized using stepwise convergent strategies. Single helical conformations have been characterized in the solid state by single crystal X-ray diffraction analysis for four of these compounds. Helices from pentameric strands 12 and 14 extend over one turn, and helices from heptameric 20 and undecameric 15 species extend to one and a half and two and a half turns, respectively. Intramolecular hydrogen bonds are responsible for the strong bending of the strands. 1H NMR shifts both in polar and nonpolar organic solvents indicate intramolecular overlap between the peripheral aromatic groups. Thus, helical conformations also predominate in solution. Molecular stochastic dynamic simulations of strand folding starting from a high energy extended linear conformer show a rapid (600 ps at 300 K) conversion into a stable helical conformation. PMID- 11486957 TI - Helical molecular programming: supramolecular double helices by dimerization of helical oligopyridine-dicarboxamide strands. AB - Helically preorganized oligopyridine-dicarboxamide strands are found to undergo dimerization into double helical supramolecular architectures. Dimerization of single helical strands with five or seven pyridine rings has been characterized by NMR and mass spectrometry in various solvent/ temperature conditions. Solution studies and stochastic dynamic simulations consistently show an increasing duplex stability with increasing strand length. The double helical structures of three different dimers was characterized in the solid phase by X-ray diffraction analysis. Both aromatic stacking and hydrogen bonding contribute the double helical arrangement of the oligopyridinedicarboxamide strand. Inter-strand interactions involve extensive face-to-face overlap between aromatic rings, which is not possible in the single helical monomers. Most hydrogen bonds occur within each strand of the duplex and stabilize its helical shape. Some inter-strand hydrogen bonds are found in the crystal structures. Dynamic studies by NMR as well as by molecular modeling computations yield structural and kinetic information on the double helices and on monomer-dimer interconversion. In addition, they reveal the presence of a spring-like extension/compression as well as rotational displacement motions. PMID- 11486958 TI - Ab initio calculations on bismuth cluster polycations. AB - Ab initio calculations on bismuth polycationic species of the types Bi(n(n-2))+, Bi(n(n-4))+, and Bi(n(n-6))+ (n = 3 - 12) were performed at the Hartree-Fock and density functional theory levels in order to investigate their general properties and the applicability of Wade's rules on bismuth polycations. Some exceptions to Wade's rules were encountered, and, moreover, several predicted and calculated minima show only meta-stable behavior. The bonding in bismuth polycations is characterized by a high degree of electron delocalization and "three-dimensional aromaticity". PMID- 11486959 TI - Is "frank" DNA-strand breakage via the guanine radical thermodynamically and sterically possible? AB - Using the reduction potential of one-electron oxidized guanosine in water and the pKa values of the radical and of the parent, the N1-H bond energy of the 2' deoxyguanosine moiety is determined to be (94.3+/-0.5) kcal mol(-1). Using the DFT method, the energy of the N1-centered guanosine radical is calculated and compared with those of the C1'- and C4'-radicals formed by H-abstraction from the 2'-deoxyribose moiety of the molecule. The result is that these deoxyribose centered radicals appear to be more stable than the N1-centered one by up to 3 kcalmol(-1). Therefore, H-abstraction from a 2'-deoxyribose C-H bond by an isolated guanosine radical should be thermodynamically feasible. However, if the stabilization of a guanine radical by intrastrand pi-pi interaction with adjacent guanines and the likely lowering of the oxidation potential of guanine by interstrand proton transfer to the complementary cytosine base are taken into account, there is no more thermodynamic driving force for H-abstraction from a deoxyribose unit. As a further criterion for judging the probability of occurrence of such a reaction in DNA, the stereochemical situation that a DNA guanosine radical faces was investigated utilizing X-ray data for relevant model oligonucleotides. The result is that the closest H-atoms from the neighboring 2' deoxyribose units are at distances too large for efficient reaction. As a consequence, H-abstraction from 2'-deoxyribose by the DNA guanine radical leading subsequently to a "frank" DNA strand break is very unlikely. The competing reaction of the guanine radical cation with a water molecule which eventually yields 8-oxo-2'-deoxyguanosine (leading to "alkali-inducible" strand breaks) has thus a chance to proceed. PMID- 11486960 TI - Pi-donation and stabilizing effects of pnicogens in carbenium and silicenium ions: a theoretical study of [C(XH2)3]+ and [Si(XH2)3]+ (X = N, P, As, Sb, Bi). AB - Quantum chemical calculations at the MP2 and CCSD(T) levels of theory are reported for cations of the general type [A(XH2)3]+ with A = C, Si and X = N, P, As, Sb, Bi. Population analysis, methyl stabilization energies (MSEs), and structural criteria were used to predict the p(pi)-donor ability of and the pi stabilization energy exerted by this series of pnicogens. All of the substituents XH2 considered in these studies invariably stabilize the triply substituted carbenium as well as the silicenium ions. The calculated data show that the intrinsic p(pi)-donation of the group 15 atoms follows the order N < P < As < Sb < Bi. However, the trend of the stabilization energies is fully reversed. The intrinsic stabilization energies of the planar carbenium ions decrease monotonically from 161.2 kcal mol(-1) for X = NH2 to 98.0 kcal mol(-1) for X = BiH2. The effective stabilization of the pnicogens in the equilibrium structures, which also includes the energy-demanding pyramidalization of the XH2 substituents, follows the same trend, although the absolute numbers are reduced to 145.6 kcalmol(-1) for X = NH2 and 53.2 kcalmol(-1) for X = BiH2. This seemingly contrasting behavior of increasing p(pi) charge donation and decreasing stabilization has already been found for other substituents. Previous studies have shown that carbenium ions substituted by chalcogens up to the fourth row also stabilize C+ less effectively with respect to heavier substituents. Of the ions investigated in this study, only the silicenium ions that are stabilized by pnicogens from the third to the sixth row of the periodic system yield increased stabilizing energies that follow the corresponding intrinsic p(pi)-donor abilities of the respective substituent. PMID- 11486961 TI - Novel ruthenium(II)2 carboxylates as catalysts for alkene metathesis. AB - The reactions of [Ru-(=CHR)Cl2(PCy3)2] (1: R = Ph; 1a: R = -CH=CPh2) with silver salts of carboxylic acids afforded new dimeric complexes of the general formula [Ru2(=CHR)2-(R'CO2)2(mu-R'CO2)2(PCy3)2(mu-H2O)] (2: R = Ph, R' = CF3; 3: R = Ph, R' = C2F5; 4: R = -CH=CPh2, R' = CF3; 5: R = Ph, R' = C6F5; 6: R = -CH=CPh2, R' = C6F5; 7: R = -CH=CPh2, R'=CCl3) in good yields. With R' = CF3, C2F5 or CCl3 these complexes are active catalysts for metathesis of acyclic alkenes, including unsaturated fatty acid esters, as well as for ring closing metathesis. The reactivity of these complexes with bases and weak donor solvents has been studied and their half-life times in several media were determined. PMID- 11486962 TI - Ferrocene-cyclam: a redox-active macrocycle for the complexation of transition metal ions and a study on the influence of the relative permittivity on the coulombic interaction between metal cations. AB - The reaction of 1,1'-ferrocene-bis(methylenepyridinium) salt with 1,4,8,11 tetraazacyclotetradecane-5,12-dione, followed by LiAlH4 reduction results in the formation of FcCyclam. Metal complexes of FcCyclam with M2+ = Co2+, Ni2+, Cu2+, and Zn2+ were synthesized from FcCyclam and the respective metal triflates. The complexation of Cu2+ and FcCyclam in CH3CN is preceeded by a rapid electron transfer, followed by a slower complex formation reaction and a reverse electron transfer. The protonation constants of FcCyclam and the stability constants for the Cu2+ complex of FcCyclam (logK = 9.26(4) for the formation of the [Cu(FcCyclam)]2+ complex) were determined in 1,4-dioxane/water 70:30 v/v, 0.1 moldm(-3), KNO3, 25 degrees C. By using FcCyclam one can selectively sense the presence of Cu2+ ions in the presence of Ni2+, Zn2+, Cd2+, Hg2+, and Pb2+ with a very large deltaE approximately 200 mV, depending on pH. The X-ray crystal structures of FcCyclam and of complexes with Co2+, Ni2+, Cu2+, and Zn2+ were determined and Fe-M2+ distances obtained: Fe-Co2+ 395.9, Fe-Ni2+ 385.4, Fe-Cu2+ 377.7, and Fe-Zn2+ 369.0 pm. The redox potential of FcCyclam is influenced in a characteristic manner by the complexation of M2+. A linear correlation of 1/r approximately/= deltaE [r = distance Fe-M2+ from crystal data, deltaE= E1/2([M(FcCyclam)]2+) - E1/2(FcCyclam)] was found; this is indicative of a mainly Coulomb type interaction between the two metal centers. The nature of the Fe...M2+ interaction was also investigated by determining deltaE in several solvents (mixtures) of different dielectric constants epsilon. The expected relation of deltaE approximately/= 1/epsilon was only found at very high values of epsilon. At epsilon < 40 increased ion-pairing appears to reduce the effective positive charge at M2+ leading to progessively smaller values of deltaE with lowered epsilon. The dependence of deltaE and epsilon can be calculated semiquantitatively by combining the Fuoss ion-pairing theory with the Coulomb model. PMID- 11486963 TI - Properties of a binaphthyl-bridged porphyrin-iron complex bearing hydroxy groups inside its cavity. AB - Hydrogen-bond formation with the terminal oxygen atom is considered to be the basis of dioxygen molecule activation by cytochrome P450. In order to verify the effect of this hydrogen bond, we have undertaken the synthesis of a model complex: a binaphthyl-bridged porphyrin bearing hydroxy groups at suitable positions ("single coronet" or "SC"). The reactivity of the iron complex of the synthesized compound towards basic ligands, dioxygen and carbon monoxide has been studied. When a bulky axial ligand such as 1-methyladamantyl-2-methylimidazole is used, only the pentacoordinated complex is obtained, and, as expected, dioxygen binds as the sixth ligand only in the cavity of the compound. Under unusually low dioxygen partial pressures and in rigorously anhydrous toluene, the pentacoordinated iron complex is completely transformed into a new species which absorbs in the visible region at 420 and 559 nm, and which we have identified as the oxygenated complex. Surprisingly, this reaction seems to occur irreversibly, based on the fact that the initial complex is not recovered after bubbling nitrogen through the solution for several hours. On the other hand, saturation of the solution with carbon monoxide transforms the complex slowly but completely into the Fe(II)-CO complex which is stable in a dioxygen-saturated toluene solution at 0 degrees C. However, by raising the temperature, it is spontaneously transformed back into the dioxygen complex; this verifies the reversibility of the dioxygen binding process. Compared with its affinity towards carbon monoxide, the SC iron complex has a much stronger affinity towards dioxygen. This remarkable property may be partly explained by hydrogen bonding between the terminal atom of the dioxygen molecule and the hydroxy groups attached to the binaphthyl bridges of the porphyrin, and also by polar neighbouring-group effects. Ligand binding and debinding constants have been determined by laser flash photolysis. Ligand-iron bond strength and hydrogen bonding have been investigated by IR and Raman spectroscopy. The role of the hydroxy groups has also been emphasized by comparing the properties of SC with those of a binaphthyl bridged porphyrin bearing methoxy groups instead of hydroxy groups. PMID- 11486964 TI - Preparation of dendritic and non-dendritic styryl-substituted Salens for cross linking suspension copolymerization with styrene and multiple use of the corresponding Mn and Cr complexes in enantioselective epoxidations and hetero Diels-Alder reactions. AB - Following work with TADDOLs and BINOLs, we have now prepared Salen derivatives (2, 3, 14, 15, 18, 19, 20, 21) carrying two to eight styryl groups for cross linking copolymerization with styrene. The Salen cores are either derived from (R,R)-diphenyl ethylene diamine (3, 15, 19, 21) or from (R,R)-cyclohexane diamine (2, 14, 18, 20). The styryl groups are attached to the salicylic aldehyde moieties, using Suzuki (cf. 1) or Sonogashira cross-coupling (cf. 11), and/or phenolic etherification (cf. 5, 7) with dendritic styryl-substituted Frechet-type benzylic branch bromides. Subsequent condensation with the diamines provides the chiral Salens. Corresponding Salens lacking the peripheral vinyl groups (cf. 12, 13, 16, 17) were also prepared for comparison of catalytic activities in homogeneous solution with those in polystyrene. Cross-linking radical suspension copolymerization of styrene and the styryl Salens, following a procedure by Itsuno and Frechet, gave beads (ca. 400 microm diameter) which were loaded with Mn or Cr (ca. 0.2 mmol of complex per g of polymer), with more than 95% of the Salen incorporated being actually accessible for complexation (by elemental analysis). The polymer-bound Mn and Cr complexes were used as catalysts for epoxidations of six phenyl-substituted olefins (m-CPBA/NMO; products 22a-f), and for dihydropyranone formation from the Danishefsky diene and aldehydes (PhCHO, C5H11CHO, C6H11CHO, products 23a-c). There are several remarkable features of the novel immobilized Salens: i) The dendritic branches do not slow down the catalytic activity of the complexes in solution; ii) the reactions with Salen catalysts incorporated in polystyrene give products of essentially the same enantiopurity as those observed in homogeneous solution with the dendritically substituted or with the original Jacobsen - Katsuki complexes; iii) some Mn loaded beads have been stored for a year, without loss of activity; iv) especially the biphenyl- and the acetylene-linked Salen polymers (p-2, -3, -20, 21, Figure 2, 3) give Mn complexes of excellent performance: after ten uses (without re-charging with Mn!) there is no loss of enantioselectivity or degree of conversion under the standard conditions. PMID- 11486965 TI - Carbon nanotube bags: catalytic formation, physical properties, two-dimensional alignment and geometric structuring of densely filled carbon tubes. AB - The catalytic CVD synthesis, using propyne as carbon precursor and Fe(NO3)3 as catalyst precursor inside porous alumina, gives carbon nanotube (CNT) bags in a well-arranged two-dimensional order. The tubes have the morphology of bags or fibers, since they are completely filled with smaller helicoidal CNTs. This morphology has so far not been reported for CNTs. Owing to the dense filling of the outer mother CNTs with small helicoidal CNTs, the resulting CNT fibers appear to be stiff and show no sign of inflation, as sometimes observed with hollow CNTs. The fiber morphology was observed by raster electron microscopy (REM), transmission electron microscopy (TEM), and atomic force microscopy (AFM). The carbon material is graphitic as deduced from spectroscopic studies (X-ray diffraction, Raman and electron energy-loss spectroscopy (EELS)). From Mossbauer studies, the presence of two different oxidation states (Fe0 and FeIII) of the catalyst is proven. Geometric structuring of the template by two different methods has been studied. Inkjet catalyst printing shows that the tubes can be arranged in defined areas by a simple and easily applied technique. Laser structuring creates grooves of nanotube fibers embedded in the alumina host. This allows the formation of defined architectures in the microm range. Results on hydrogen absorption and field emission properties of the CNT fibers are reported. PMID- 11486966 TI - Synthesis of functionalized pyrrole and indole derivatives through carbometallation of lithiated double bonds. AB - Bis(2-lithioallyl)amines derived from bis(2-bromoallyl)amines undergo intramolecular carbometallation of a lithiated double bond, giving dilithiated dihydropyrroles. The cyclizations are promoted by N,N,N',N' tetramethylethylenediamine (TMEDA). Reaction of these intermediates with electrophiles allows the preparation of some new fused and nonfused five-membered functionalized heterocycles. Although 2-lithioallylamines do not suffer intermolecular carbometallation, dimerization products are obtained with their copper or zirconium derivatives. Finally, the application of this new reaction to 2-lithio-N-(2-lithioallyl)anilines leads to 3-lithiomethylindole derivatives, which are transformed to functionalized indole derivatives by reaction with electrophiles. PMID- 11486967 TI - Palladium catalyst systems for cross-coupling reactions of aryl chlorides and olefins. AB - A detailed investigation into the influence of phosphines, additives, bases and solvents on the Heck coupling reaction of 4-trifluoromethyl-1-chlorobenzene (2) is presented. It is shown that a number of catalyst systems exist for efficient cross coupling of electron-deficient aryl chlorides with various olefins. Basicity and steric demand of the ligand are two factors which determine the success of the reaction. In addition the phosphine/palladium ratio, the correct type and amount of additive, and finally the use of an appropriate base and solvent are also important. The optimised reaction conditions are applied for the arylation of styrene, 2-ethylhexyl acrylate and N,N-dimethyl acrylic amide with various aryl chlorides. PMID- 11486968 TI - The protonation of gaseous cyclopropane. AB - The protonation of cyclopropane by gaseous Bronsted acids of varying strength in radiolytic experiments at atmospheric pressure leads to two distinct C3H7- isomers that have been sampled by their reaction with benzene. The neutral end products, nC3H7-C6H5 and iC3H7-C6H5, arise from the electrophilic aromatic substitution reaction with the cC3H7+ and iC3H7+ ions, respectively. Their relative abundance was studied as a function of pressure, temperature, and the presence of additives in the gaseous systems; the results indicate a large extent of isomerization to the thermodynamically favored iC3H7+ from the protonation by strong acids. The presence of a kinetic barrier prevents any thermal isomerization from taking place in the time frame of 10(-8) s. In the peculiar case in which protonated benzene is the Bronsted acid, C3H7+ ions are formed in the presence of neutral benzene within the same ion - molecule complex. The ensuing reaction shows that cC3H7+ ions are formed exclusively and react in the 10(-10) s(-1) estimated lifetime of the complex. Still, such cC3H7+ ions undergo complete randomization of their hydrogen atoms; this points to a low kinetic barrier for the process. Agreement is found between the reported experimental results and updated computations of the relevant species in the C3H7+ potential energy surface. PMID- 11486969 TI - Solid-phase organometallic synthesis. AB - A solid-phase synthesis approach for a class of molybdenum carbonyl complexes has been developed. The system can be used to perform metal-complexation, ligand substitution reactions and oxidative eliminations on the solid phase and to cleave the final complexes under mild and selective conditions. Comparison is made to corresponding soluble complexes and liquid-phase reactions. PMID- 11486970 TI - Mapping concentration profiles within the diffusion layer of an electrode: application to redox catalysis. AB - Catalytic reductions of some aromatic halides were performed at a millimetric electrode with several redox mediators. The resulting concentration profiles were monitored amperometrically by placing an ultramicroelectrode inside the diffusion layer produced at the former electrode. The features of redox catalysis and the subsequent structuring of the diffusion layer were investigated experimentally under steady-state conditions imposed by the spontaneous convection of the solution. The concentration profiles established from the probe measurements were in agreement with our theoretical predictions, based on fast kinetics of redox catalysis. Under these conditions, very similar to preparative electrosynthesis, the diffusion layer separates into two domains where pure diffusion takes place and the concentration profiles therein are mainly linear. We demonstrate that the limit between these two zones does not depend on kinetics, but is rather fixed by the product of the ratio of the bulk concentrations of each species and the ratio of their diffusion coefficients. PMID- 11486971 TI - Synthesis of lipidated eNOS peptides by combining enzymatic, noble metal- and acid-mediated protecting group techniques with solid phase peptide synthesis and fragment condensation in solution. AB - Lipid-modified proteins play decisive roles in important biological processes such as signal transduction, organization of the cytoskeleton, and vesicular transport. Lipidated peptides embodying the characteristic partial structures of their parent lipidated proteins and semisynthetic proteins synthesized from such peptides are valuable tools for the study of these biological phenomena. We have developed an efficient synthesis strategy that allows for the synthesis of long multiply lipidated peptides embodying various side chain functional groups. The strategy was successfully applied in the synthesis of the N-terminal undetrigintapeptide of endothelial NO-synthase and related lipopeptides. Key elements of the synthesis strategy are the combined use of the enzyme-labile para phenylacetoxybenzyloxycarbonyl (PhAcOZ) urethane as N-terminal blocking group, the Pd0-sensitive allyl ester as C-terminal protecting function and acid-labile side chain protecting groups for solution-phase synthesis of labile S palmitoylated building blocks under the mildest conditions with solid-phase techniques and solution-phase fragment condensations. The successful synthesis of the triply lipidated 29-mer eNOS peptide convincingly demonstrates the full capacity of the protecting group methods. PMID- 11486972 TI - HIV/AIDS: twenty years later and the battle still rages. PMID- 11486973 TI - Increasing awareness of health care concerns during Ramadan. PMID- 11486974 TI - Guiding principles for drug use at the patient and population levels. PMID- 11486975 TI - To link or not to link: pharmaceutical care and medication dispensing. PMID- 11486976 TI - Perspectives on generic pharmaceuticals: a series in JAPhA. PMID- 11486977 TI - American pharmaceutical education, 1902-1952. PMID- 11486978 TI - Informed consent: what every pharmacist should know. PMID- 11486979 TI - Using the Internet to enhance pharmacy-based patient care services. PMID- 11486980 TI - Improving drug therapy for patients with asthma--part 1: Patient outcomes. AB - OBJECTIVE: To evaluate the effects of a therapeutic outcomes monitoring (TOM) program on selected process and outcome measures. DESIGN: Prospective, controlled, multicenter study. SETTING: Community pharmacies throughout Denmark (16 intervention, 15 control). PATIENTS: Five hundred patients with asthma aged 16 to 60 years and treated in primary care. INTERVENTION: TOM is a community based program for pharmaceutical care. Using a structured, seven-step, cyclical outcome improvement process, TOM pharmacists identify and resolve (or refer) problems with drug therapy that, if not addressed, might result in therapeutic failure or adverse effects. Equal emphasis is placed on the patient's perspective (e.g., coping, control, and empowerment) and the professional's perspective (e.g., adherence, patient knowledge, and therapeutic problems). TOM requires cooperation among pharmacists, patients, and physicians. MAIN OUTCOME MEASURES: Asthma symptom status, days of sickness, health-related and asthma-specific quality of life, use of health care services and resources, and satisfaction with health care and pharmacy. INTERMEDIATE OUTCOME AND PROCESS MEASURES: Peak expiratory flow rate (PEFR), knowledge of asthma and asthma medications, inhalation errors, and drug therapy problems in the TOM group. RESULTS: The mean individual differences for TOM and control patients were tested. Beneficial effects were found for the following outcome measures: asthma symptom status, days of sickness, and health-related and asthma-related quality of life. Satisfaction with health care and pharmacy varied throughout the course of the project, with no significant difference between groups at the final evaluation. Although not statistically significant, differences in use of services were considered to be clinically significant and encouraging. Beneficial effects were found for knowledge of asthma and medications, inhalation errors, drug use and drug therapy problems. No significant differences were found for PEFR. CONCLUSION: The project demonstrated that therapeutic outcomes monitoring by community pharmacists is an effective strategy for improving the quality of drug therapy for asthma patients in primary health care. PMID- 11486981 TI - Improving drug therapy for patients with asthma-part 2: Use of antiasthma medications. AB - OBJECTIVES: To describe the use of antiasthma drugs among the study patients and to evaluate whether therapeutic outcomes monitoring (TOM) is associated with improved quality of drug therapy. DESIGN: Prospective, controlled, multicenter study. Consumption of antiasthma medications was measured as the number of defined daily doses (DDDs) purchased. Data were collected from the pharmacies' computer systems for a period beginning 6 months before the start of the study (period 1) and during its first and second half-years (periods 2 and 3). Treatment changes for TOM patients were classified on the basis of drug regimens at periods 1 and 3. SETTING: Community pharmacies in Denmark (16 intervention, 15 control). PATIENTS: Five hundred patients with asthma aged 16 to 60 years who were being treated in primary health care; this study used data from 350 patients from this sample. INTERVENTION: TOM. MAIN OUTCOME MEASURES: Changes in the use of individual drugs and changes in therapeutic patterns--distribution of purchased drugs; proportion of corticosteroid users; frequency of drug regimens used; treatment changes for TOM patients. RESULTS: TOM patients' consumption of beta2 agonists decreased by 12% overall from period 1 through period 3, while control patients' consumption of these medications decreased by only 1%. TOM patients' use of inhaled corticosteroids increased by more than 50% compared with 9% among controls. In both groups, about one-half of all purchased DDDs were for inhaled beta2-agonists. The proportion of inhaled corticosteroids increased from 27% to 42% of total DDDs for the TOM group and remained constant for controls. Of patients using beta2-agonists, 68% also used inhaled steroids initially in both the TOM and control groups. The proportion of inhaled steroid users in the TOM group increased to 84%, and to 70% among controls. The most common regimen was inhaled short-acting beta2-agonists and corticosteroids in combination, and the second most common regimen was monotherapy with short-acting beta2-agonists. With time, the regimens changed more toward consensus guidelines among TOM patients. Changes in drug therapy totaled 451, averaging 2.4 changes per TOM patient. The largest number of changes (49%) involved inhaled corticosteroids. CONCLUSION: Changes in medication use among TOM patients were toward improved asthma treatment. Our results show that community pharmacists, physicians, and patients, working together, can improve prescribing, solve drug therapy problems, and improve outcomes for patients with moderate-to-severe asthma. PMID- 11486982 TI - Comparison of German St. John's wort products according to hyperforin and total hypericin content. AB - OBJECTIVES: To compare the hyperforin and hypericin content of currently available St. John's wort products and to determine their batch-to-batch reproducibility. DESIGN: Representative products were obtained either directly from the manufacturer or purchased from pharmacies in and around Frankfurt, Germany. For five batches from each of the eight manufacturers, 10 individual dosage forms (tablets or capsules) were analyzed for both hyperforin and hypericin content. SETTING: Laboratories of the Institute of Pharmaceutical Chemistry at Johann Wolfgang Goethe University, Frankfurt, Germany. PRODUCTS: Eight German St. John's wort products containing from 250 mg to 612 mg dry extract were studied. Three of these products are capsules, four are film-coated tablets, and one is a sugar-coated tablet. Two of the products (Jarsin 300 and Neuroplant 300) are also available in the United States. METHODS: Hyperforin concentrations were analyzed by high-performance liquid chromatography. Total hypericin concentrations were determined by polarography, an electrochemical method. Concentrations were compared among different batches of the same product and among products from different manufacturers. RESULTS: The products contained widely differing amounts of hypericin and hyperforin, even after correcting for differences in the amount of extract per dose. Some products demonstrated consistent concentrations of hyperforin and hypericin from batch to batch, others exhibited pronounced interbatch variability. CONCLUSION: The St. John's wort preparations studied exhibited large differences in hypericin and hyperforin content and are not interchangeable for the treatment of mild-to-moderate depression. Pharmacists should take this variability into account when counseling patients on the use of St. John's wort products. PMID- 11486983 TI - Historical overview of generic medication policy. AB - OBJECTIVE: To provide a historical perspective on controversies surrounding the use of generic drugs. DATA SOURCES: Articles were indexed initially using terms such as generic medications, generic drugs, multisource medications, and multisource drugs. These terms were used to search indexing services such as MEDLINE, International Pharmaceutical Abstracts, CINAHL (a database of nursing and allied health literature), Science Citation Index, Psychological Abstracts, and Wilson Indexes to Journal Articles. STUDY SELECTION: Performed by the authors, with preference given to events from 1951 to the present. DATA EXTRACTION: Not applicable. DATA SYNTHESIS: The history of generic drug use is a history of conflict from a variety of perspectives. The primary conflict is economic, in which manufacturers of brandname pharmaceuticals aggressively seek to protect their patents from a variety of groups (e.g., the federal government, managed care organizations, consumer groups) that want access to less expensive medications. Another conflict is professional, especially for the members of the pharmacy profession who view drug product selection as an important opportunity for pharmacists to use their professional judgment. The most confusing conflict is the scientific discussion of bioequivalence and product quality. The brand manufacturers suggest that not all products are bioequivalent and of the same quality. This position has been opposed by the pharmacy profession, generic drug manufacturers, health care institutions, and the Food and Drug Administration. CONCLUSION: Generic drug use has increased dramatically during the past 50 years and is an accepted part of health care. However, the economic consequences of generic drug use are sufficiently high for this activity to continue as a source of controversy in the future. PMID- 11486984 TI - Economics and structure of the generic pharmaceutical industry. AB - OBJECTIVE: To review key economic trends of the generic medication market and analyze the changing structure of the generic medication industry. DATA SOURCES: Articles were indexed initially using terms such as generic medications, generic drugs, multisource medications, and multisource drugs. These terms were used to search indexing services such as MEDLINE, International Pharmaceutical Abstracts, CINAHL (a database of nursing and allied health literature), Science Citation Index, Psychological Abstracts, and Wilson Indexes to Journal Articles. STUDY SELECTION: Performed by the authors. DATA EXTRACTION: Not applicable. DATA SYNTHESIS: The generic pharmaceutical market has the potential to play an important role in containing drug costs, although the amount that could be saved through the use of generic medications is not easy to measure. If estimates for the future growth in the use of generic products prove correct, the proportion of pharmaceutical sales attributable to generic products will remain in the 9% to 10% range through the first decade of the 21st century. The generic pharmaceutical industry includes several categories of companies based on business strategies. Further consolidation is expected as more resources are needed to address patent challenges stimulated by the Drug Price Competition and Patent Restoration Act. Companies are also entering into partnerships with research manufacturers to share profits from proprietary products. Future growth of the generic market will require more than increased use of generic products. Therapeutic interchange, involving switching from single-source, patent-protected products to products within the therapeutic class for which there are generic equivalents, represents a major growth opportunity for generic drug manufacturers because the savings potential is more significant than that for straightforward substitution of generic for brandname products. CONCLUSION: As it responds to challenges and opportunities, the generic pharmaceutical industry will continue to be a major force shaping the economics of medication use. PMID- 11486985 TI - Prevention and management of colorectal cancer in women. AB - OBJECTIVE: To review prevention and management strategies for colorectal cancer, with an emphasis on studies pertaining to women. DATA SOURCES: Articles published from January 1990 through February 2001 identified through a MEDLINE search using the term colorectal cancer and the additional terms screening, prevention, and treatment. Additional references were identified from the bibliographies of the retrieved articles. DATA SYNTHESIS: Colorectal cancer is the third most common non-skin cancer in women, after breast and lung cancers. Many women underestimate their risk of colorectal cancer, which may lead them to underuse screening measures that have been proven to reduce disease morbidity and mortality. For average-risk women and men > or = 50 years of age, pharmacists should recommend regular screening for early detection and prevention of colorectal cancer. In its earliest, most curable stages, colorectal cancer is often asymptomatic. However, patients who report signs and symptoms, such as blood in the stool, abdominal pain, changes in bowel habits, unexplained weight loss, or iron deficiency anemia, should be referred for medical evaluation. The use of chemopreventive agents for colorectal cancer, such as nonsteroidal anti-inflammatory drugs, hormone replacement therapy, and dietary calcium, holds significant promise, but further studies are needed before these agents can be recommended for cancer prevention in the general population. Surgical resection is the primary treatment modality for colorectal cancer, and adjuvant chemotherapy is recommended in patients with stage III disease and some high-risk patients with stage II disease. Pharmacists should be aware that women are more susceptible to dose related toxicity effects of fluorouracil and leucovorin combination chemotherapy, the first-line regimen for adjuvant chemotherapy. CONCLUSION: Although often perceived as a disease that primarily affects men, colorectal cancer is an equally important health concern for women. By providing education and counseling, pharmacists can help raise women's awareness of this disease and encourage them to take steps to significantly reduce their risk. PMID- 11486986 TI - New guidelines for managing hypercholesterolemia. AB - OBJECTIVE: To summarize for pharmacists the Adult Treatment Panel III (ATP III), recently issued guidelines for managing hypercholesterolemia, from the National Cholesterol Education Program (NCEP). DATA SOURCES: Executive summary of ATP III, and other pertinent literature as determined by the author. STUDY SELECTION: Not applicable. DATA EXTRACTION: By the author. DATA SYNTHESIS: Like previous guidelines issued by NCEP, ATP III focuses on lowering of low-density lipoprotein cholesterol (LDL-C) as a primary focus and using exercise, diet, and pharmacotherapy as a primary means of lowering patients' coronary heart disease (CHD) risks. The new guidelines recognize LDL-C levels of less than 100 mg/dL as optimal for all patients, and increase attention on high triglyceride levels (above 200 mg/dL). ATP III places more emphasis on identifying patients at risk for CHD and CHD events (e.g., myocardial infarctions, revascularization procedures). To apply the recommendations of ATP III in pharmaceutical care practice, pharmacists should follow a six-step process: (1) Assess the patient's lipid profile (full panel, not just total cholesterol); (2) assess and categorize the patient's CHD risk (using a point system reflecting the levels of risk inherent in certain factors); (3) establish treatment goals and approaches (the greater the risk, the more aggressive the management); (4) initiate therapeutic lifestyle changes (including new recommendations for low intake of saturated fats and dietary cholesterol); (5) initiate LDL-C lowering drug therapy (often with combination therapy); and (6) consider other lipid factors (particularly hypertriglyceridemia and the metabolic syndrome). CONCLUSION: Most patients who begin lipid-lowering therapy stop it within 1 year, and only about one-third of patients reach treatment goals. The release of the ATP III guidelines provides pharmacists a great opportunity to enhance pharmaceutical care services directed specifically at patients with hyperlipidemia. PMID- 11486987 TI - An organizational tool to describe and evaluate group performance in the health care setting. AB - OBJECTIVE: To develop and test a tool for evaluating the performance of working groups in the institutional health care setting and to discuss its utility for pharmacists and other health care providers. DESIGN: A model for understanding how groups make decisions in health care settings was developed based on theories derived from organizational research. Our general group decision-making model was based on data collected from a national survey of members of hospital pharmacy and therapeutics (P&T) committees. Members were asked to describe how they interacted when making decisions regarding whether to add or delete drugs from their hospital's formulary. The relationships were examined using structural equation modeling. SETTING: Large teaching hospitals. PARTICIPANTS: Key members of P&T committees. MAIN OUTCOME MEASURES: Committee members' responses about committee structure, organizational environment, quality of their interactions with each other, and the committee's decision-making abilities. Individual responses were aggregated into group measures to improve reliability and validity. RESULTS: Responses were received from 114 of the 222 hospitals surveyed (51.3%), and analysis was conducted on 95 hospitals (42.8%) for which three or more group members responded. Positive interactions among group members had the greatest influence on the group's decision-making abilities (beta = 0.92). These interactions were facilitated by a well-designed group (gamma = 0.71) and the support of the environment within which the committee operates (gamma = 0.29). CONCLUSION: The general group decision-making model tested in this study may be useful in understanding how the internal processes of health care groups such as P&T committees influence group performance and, hence, group outputs. Health care providers who apply a systematic approach to assessing group performance will find this model useful because it provides a starting point for examining the process, a way to show interrelationships among various group activities, and a guide to which strategies may be most effective for improving the decision-making process. PMID- 11486988 TI - Pharmacy through the ages. Taraxacum officinale. PMID- 11486989 TI - Osteoporosis: helping your patients avoid a bad break. PMID- 11486990 TI - Compliance action program: an opportunity to fight the nation's leading cause of death. PMID- 11486991 TI - Prioritizing vaccines: whom to immunize first? PMID- 11486992 TI - Pharmacogenomics: opening new vistas in pharmacotherapy. PMID- 11486993 TI - Drug information centers on the Internet. PMID- 11486994 TI - Antithrombotic strategy in stroke. AB - Numerous randomised controlled trials have been devoted to antithrombotic strategy in stroke, thus making evidence-based recommendations possible. The use of antithrombotic drugs is crucial in the treatment of ischemic stroke though often limited by the inherent risk of intra-cerebral bleeding. In the prevention of stroke, the strategy depends on the underlying etiology: (i) antiplatelet drugs (with aspirin as first choice) in atherothrombotic stroke, and (ii) oral anticoagulants in cardioembolic stroke. In the acute treatment, the strategy depends on whether IV rt-PA can be performed; if rt-PA is available and approved, its use is recommended within 3 h of the onset of symptoms provided there is strict adherence to the inclusion and exclusion criteria. In all other cases, aspirin is the treatment of choice, associated with low dose LMWH in the event of restricted mobility. There is no evidence for efficacy of high dose heparin (or LMWH) in stroke, except in cerebral venous thrombosis. PMID- 11486995 TI - Thrombophilia, thrombosis and pregnancy. AB - The risk of venous thromboembolism (VTE) in pregnancy is 0.05-1.8%, six times greater than in the non-pregnant state, and pulmonary embolism remains the most common cause of maternal death. Maternal age, previous history of VTE, Caesarean section and the presence of thrombophilia, significantly increase the risk of VTE. Acquired or hereditary thrombophilia occur in almost two-thirds of women presenting with recurrent miscarriages, pre-eclampsia, intrauterine growth restriction, abruptio placentae, or stillbirth, which are associated with microvascular thrombosis in placental blood vessels. Women with VTE during pregnancy and especially those with thrombophilia require individualized management, based on the type of defect, the family history and the presence of additional risk factors. These factors are important in determining the dose and duration of antithrombotic therapy during pregnancy and the puerperium, and the thromboprophylactic strategy for future pregnancies. Oral anticoagulants are now seldom used during pregnancy because of their significant side effects. Low molecular-weight heparins (LMWHs) are increasingly replacing unfractionated heparin in the prevention and treatment of VTE during pregnancy. LMWHs have also been shown to be effective in improving the outcome of pregnancy in women with previous obstetric complications. PMID- 11486996 TI - Oral contraceptives, hormone replacement therapy and thrombosis. AB - Oral contraceptives and hormone replacement therapy are used by hundreds of millions of women worldwide. Since the early 1960s it is known that female hormones increase the risk of venous thrombosis, myocardial infarction and stroke. This risk is still present with current low-dose oral contraceptives and, even though in absolute terms the risk is small, oral contraceptives form the major cause of thrombotic disease in young women. The risk is higher during the first year of use (up to I per 1000 per year), with the use of desogestrel- or gestodene-containing oral contraceptives ("third generation progestogens") and among women with a prothrombotic predispositon. Hormone replacement therapy increases the risk of venous thrombosis, while results of randomised trials so far do not substantiate the expectation of a beneficial effect on the risk of arterial cardiovascular disease. First results are emerging that specific subgroups of women, with prothrombotic or other abnormalities, may be at risk, especially during the first years of use of hormone substitution. PMID- 11486997 TI - Protease crosstalk with integrins: the urokinase receptor paradigm. AB - Migratory cells use both adhesion receptors and proteolytic enzymes to regulate their interaction with and response to extracellular matrices. Cooperation between integrins and proteases operates at several levels: integrin signaling induces proteases, proteases co-localize with integrins, and proteases regulate the interface between integrins and the intracellular cytoskeleton. One protease system intimately connected to integrins is the urokinase/urokinase receptor(uPAR)/plasmin system. Recent studies indicate urokinase promotes the ligand-like binding of its receptor to a set of beta1 and beta2 integrins, this binding in turn affecting integrin signaling and cell migration. The glycolipid anchor of uPAR associates with cholesterol-rich membrane rafts. Binding of uPAR to integrins may enrich integrin clusters with signaling molecules such as src family kinases that localize to rafts and are important to integrin function. Signals derived from integrin/uPAR complexes promote the function of other integrins. Thus the urokinase/plasmin system coordinates with integrins to regulate cell: matrix interactions. PMID- 11486998 TI - Genetic interactions between the coagulation and fibrinolytic systems. AB - Nearly all of the genes encoding the established coagulation and fibrinolytic factors have been successfully altered or disrupted in transgenic mice. Although comprehensive studies of each of these gene-targeted mouse lines are still ongoing, the initial findings have significantly refined our understanding of the roles of selected hemostatic factors in vivo, and occasionally altered long standing concepts. This review summarizes some of the progress that has been made in the generation and phenotypic characterization of mice lacking key hemostatic factors, including coagulation, fibrinolytic, platelet and endothelial cell associated factors. New insights regarding the role(s) and interplay of hemostatic factors that have emerged from detailed studies of mice carrying multiple deficits in coagulation and fibrinolytic system components are highlighted. PMID- 11486999 TI - Tissue plasminogen activator in nervous system function and dysfunction. AB - The extracellular protease tissue plasminogen activator (tPA) has been implicated in various normal and pathological situations in the mammalian nervous system. The availability of (i) transgenic and knock-out mice in which the expression level of tPA can be widely varied, (ii) in vivo models for studying function and disease, and (iii) culture models for examining cell behavior, has allowed a detailed evaluation of many of these proposed functions. This chapter summarizes the current state of knowledge of possible roles for the tPA/plasminogen system in neuronal function and dysfunction. PMID- 11487000 TI - The conformational basis of thrombosis. AB - Antithrombin readily undergoes a spontaneous transition from its active five stranded form to a six-stranded inactive latent form. The recognition of this change in plasma has been obscured by the immediate linkage of newly formed latent antithrombin to a molecule of active antithrombin to give a dimer with an electrophoretic mobility readily confused with that of native active antithrombin. A new micromethod now allows unequivocal identification of latent antithrombin in whole plasma. This shows that at 37 degrees C some 10% of plasma antithrombin is converted to the latent form in 24 h. The rate of conversion is greatly accelerated at increased temperatures, as occurs in the pasteurisation of plasma concentrates that should now be checked for efficacy. But increased transition also occurs in the plasma at the slightly increased temperatures that accompany incidental infections. This is of particular significance if there is a conformationally unstable variant of antithrombin; here fever can provoke a sudden transition with the onset of a characteristically severe episode of thromboembolism. Such variants are not rare and include those previously classified as pleiotropic. The precise structural pathway, now known with antithrombin, provides a model of the changes occurring in other conformational diseases, including Alzheimer's and the prion dementias. PMID- 11487001 TI - Getting at the variable expressivity of von Willebrand disease. AB - Von Willebrand disease (VWD) is a heterogeneous bleeding disorder caused by abnormalities of von Willebrand factor (VWF). VWF levels vary widely in the general population, and this variation is likely to be a major factor accounting for the incomplete penetrance and variable expressivity of VWD. In addition, variation in VWF level may play an important role in determining the risk of venous thrombosis. A large component of the variation in VWF level in the general population has been shown to be attributable to genetic factors. This review will focus on the current understanding of the genetic causes for variation in VWF level, and will highlight future directions for getting at the variable expressivity of von Willebrand disease. PMID- 11487002 TI - Treatment of von Willebrand disease. AB - In von Willebrand disease, there are two main options for the treatment of spontaneous bleeding episodes and for bleeding prophylaxis: desmopressin and transfusional therapy with plasma products. Desmopressin is the treatment of choice for most patients with type 1, who account for approximately 70 to 80 per cent of all cases with the disease. This non-transfusional hemostatic agent raises endogenous factor VIII and von Willebrand factor three- to fivefold and thereby transiently corrects both the intrinsic coagulation and primary hemostasis defects. In patients with the more severe type 3 and in the majority of those with type 2 desmopressin is not effective or is contraindicated, so that it is usually necessary to resort to plasma concentrates containing factor VIII and von Willebrand factor. Concentrates treated with virus inactivation methods should be preferred to cryoprecipitate because they are equally effective and perceived as safer. PMID- 11487003 TI - The molecular basis of inherited afibrinogenaemia. AB - This article reviews the substantial progress made in understanding the molecular basis of inherited afibrinogenaemia (or congenital afibrinogenaemia), an autosomal recessive disorder characterised by the complete absence of detectable fibrinogen. The identification in 1999 of the first genetic defect, recurrent homozygous deletions of approximately 11 kb of the fibrinogen alpha-chain (FGA) gene, revealed that the disease was caused by defective fibrinogen synthesis, and led to the subsequent analysis of the three fibrinogen genes in other affected individuals with the identification of numerous causative mutations. Combined analyses of more than thirty unrelated afibrinogenaemia families from various ethnic groups have shown that the majority of patients have truncating mutations in the FGA gene although intuitively all three fibrinogen genes might be equally implicated. These results will facilitate molecular diagnosis of the disorder, permit prenatal diagnosis for families who so desire, and pave the way for new therapeutic approaches such as gene therapy. PMID- 11487004 TI - Von Willebrand factor propeptide in vascular disorders. AB - Von Willebrand factor (VWF) is a multifunctional plasma protein that plays a prominent role in haemostasis. In endothelial cells, processing of its precursor pro-VWF results in the formation of two large polypeptides, mature VWF and a propeptide. These proteins are co-secreted on an equimolar basis but are cleared from the circulation at different rates. VWF levels are frequently elevated in response to vascular disorders. Similarly, propeptide levels are increased under these conditions, although primarily in fulminant vascular disease, such as thrombotic thrombocytopenic purpura and septicemia. In chronic vascular disease, e.g. diabetes or peripheral vascular disease, propeptide levels are much less elevated. The differential response of VWF and propeptide levels to vascular disease could provide a means to assess the extent and time course of endothelial cell activation. After secretion, the propeptide may play a role in modulating cellular adhesion processes. Thus, enhanced propeptide secretion seems not to be of merely diagnostic significance. PMID- 11487005 TI - Gene therapy in hemophilia: clinical trials update. AB - Disorders caused by inborn genetic errors have been a primary target for treatment by gene transfer. Hemophilia A and B have been considered especially important targets because the genes for factor VIII and IX are well characterized, levels of factor VIII and IX do not require complex regulation, small increases in factor level would have significant clinical benefits, good clinical and laboratory tests of efficacy exist, and excellent animal models of hemophilia are available. Four clinical trials of gene transfer in hemophilia, two in hemophilia A and two in hemophilia B, are currently underway or have been completed and two other trials have been approved. The collective interim results from these trials indicate that the current approaches and doses are safe and that low levels of expression are detected. These studies support the continued development of gene transfer as a potential treatment option for hemophilia. PMID- 11487006 TI - The vascular biology of the glycoprotein Ib-IX-V complex. AB - It has long been recognized that binding of von Willebrand factor (vWf) by the platelet membrane glycoprotein (GP) Ib-IX-V complex initiates the cascade of events leading to thrombosis and haemostasis. In the last several years, however, it has become apparent that the GP Ib-IX-V complex plays a much broader role in vascular biology, a recognition in part due to the identification of novel adhesive ligands for GP Ib-IX-V such as P-selectin and Mac-1. Binding of vWf by the GP Ib-IX-V complex is also important in formation of the developing thrombus. Platelets can utilize the GP Ib-IX-V complex and PSGL-1 to roll on activated endothelium via surface-expressed endothelial P-selectin. The GP Ib-IX-V complex can bind the I domain of Mac-1, a binding interaction potentially relevant to transmigration of macrophages through mural thrombus, a process required for vessel remodeling post angioplasty. This review will focus on recent advances in our understanding of the structure and function of this important platelet receptor, with particular emphasis on insights made within the last two years. PMID- 11487007 TI - Platelet collagen receptors. AB - Collagens are important platelet activators in the vascular subendothelium and vessel wall. Since the regulation of platelet activation is a key step in distinguishing normal haemostasis from pathological thrombosis, collagen interactions with platelets are important targets for pharmacological control. Platelets have two major receptors for collagens, the integrin alpha2beta1, with a major role in adhesion and platelet anchoring and the Ig superfamily member, GPVI, principally responsible for signalling and platelet activation. In addition, GPIb-V-IX, can be considered as an indirect collagen receptor acting via von Willebrand factor as bridging molecule and is essential for platelet interactions with collagen at high shear rates. There is some evidence for additional receptors, which may regulate the response to individual collagen types. This review discusses how these receptors work separately with specific agonists and proposes possible mechanisms for how they work together to regulate platelet activation by collagen, which remains controversial and poorly understood. PMID- 11487008 TI - Cytoskeletal proteins and platelet signaling. AB - The actin filament network fills the cytoplasm of unstimulated platelets and connects with a submembranous latticework of short cross-linked actin filaments, known as the membrane skeleton. One function of the cytoskeleton is to direct the contours of the membrane in the unstimulated platelet and the rapid changes in shape in the activated platelet. Activation-induced changes result from events such as phosphorylation or calpain-induced cleavage of cytoskeletal proteins. The specific reorganizations depend upon the combination of signals to which platelets are exposed. A second function of the cytoskeleton is to bind other cellular components; it binds signaling molecules, localizing them to specific cellular locations; it binds the plasma membrane regulating properties of the membrane, maintaining microdomains in the membrane, or regulating activities of membrane proteins. In this way, the cytoskeleton plays a critical role in regulation of spatial organizations and, thus, in the integration of cellular activities. PMID- 11487009 TI - A journey with platelet P-selectin: the molecular basis of granule secretion, signalling and cell adhesion. AB - P-selectin is a transmembrane protein that resides within the alpha granule membrane of unstimulated platelets. The "extracellular" domains face into the lumen of the granule and the cytoplasmic tail extends into the platelet cytoplasm. Upon platelet stimulation, P-selectin is phosphorylated and translocated to the plasma membrane via a secretory pathway. P-selectin in the plasma membrane surface is exposed and serves as a cell adhesion receptor to interact with other cell receptors, including PSGL-1 and GPIb. P-selectin upregulates tissue factor in monocytes and leads to leukocyte accumulation in areas of vascular injury associated with thrombosis and inflammation. PMID- 11487010 TI - ADP receptors of platelets and their inhibition. AB - ADP plays a crucial role in haemostasis and thrombosis and its receptors are potential targets for antithrombotic drugs. Two G-protein coupled P2 receptors contribute to platelet aggregation: the P2Y1 receptor initiates aggregation through mobilisation of calcium stores, while the more recently identified P2Y12 receptor coupled to adenylyl cyclase inhibition is essential for a full aggregation response to ADP and the stabilisation of aggregates. The latter is defective in certain patients with a selective congenital deficiency of aggregation to ADP. It is also the target of the antithrombotic drug clopidogrel and of ATP analogues and other compounds currently under evaluation. In addition, the P2X1 ionotropic receptor is present in platelets but its role is not yet completely known. Studies in P2Y1 knock-out mice and experimental thrombosis models using selective P2Y1 antagonists have shown that the P2Y1 receptor, like the P2Y12 receptor, is a potential target for new antithrombotic drugs. PMID- 11487011 TI - Angiogenesis research: guidelines for translation to clinical application. AB - Angiogenesis research is being translated to the clinic. Certain guidelines may facilitate this effort. Recruitment of endothelial cells by a tumor is an early event in angiogenesis, a process regulated at genetic and epigenetic levels. The microvascular endothelial cell has become an important second target in cancer therapy. Angiogenesis inhibitors are either "direct" or "indirect" and their optimal dosing depends on a different logic than conventional chemotherapy. Conversely, antiangiogenic scheduling of chemotherapy can by-pass drug resistance. Like all solid tumors, hematologic malignancies are angiogenesis dependent. Further, angiogenesis is modulated by proteins and cells from the hematopoietic and hemostatic systems. Clinical testing of angiogenesis inhibitors has accentuated the need for surrogate markers of tumor angiogenesis activity. Microvessel density, so valuable as a prognostic indicator of metastatic risk, cannot determine efficacy of an angiogenesis inhibitor. In the future, angiogenesis inhibitors may be added to chemotherapy or to radiotherapy, or to other modalities. Also, combinations of angiogenesis inhibitors may be administered together. PMID- 11487012 TI - Hermansky-Pudlak syndrome and Chediak-Higashi syndrome: disorders of vesicle formation and trafficking. AB - The rare autosomal recessive metabolic disorders Hermanky-Pudlak syndrome (HPS) and Chediak-Higashi syndrome (CHS)share the clinical findings of oculocutaneous albinism and a platelet storage pool deficiency. In addition, HPS exhibits ceroid lipofuscinosis and CHS is characterized by infections and an accelerated phase. The two disorders result from defects in vesicles of lysosomal lineage. Of the two known HPS-causing genes, HPS1 has no recognizable function, while ADTB3A codes for a subunit of an adaptor complex responsible for new vesicle formation from the trans-Golgi network. Other HPS-causing genes are likely to exist. The only known CHS-causing gene, LYST, codes for a large protein of unknown function. In general, HPS appears to be a disorder of vesicle formation and CHS a defect in vesicle trafficking. These diseases and their variants mirror a group of mouse hypopigmentation mutants. The gene productsinvolved will reveal how the melanosome, platelet dense body, and lysosome are formed and trafficked within cells. PMID- 11487013 TI - Beta3 tyrosine phosphorylation in alphaIIbbeta3 (platelet membrane GP IIb-IIIa) outside-in integrin signaling. AB - The platelet integrin alphaIIbbeta3 not only binds fibrinogen and von Willebrand factor to mediate platelet aggregation and adhesion, it also serves as a signaling receptor. Platelet agonists such as ADP, thrombin and collagen induce "inside-out" signaling which activates the receptor function of alphaIIbbeta3 for soluble fibrinogen. Subsequent platelet aggregation leads to "outside-in" signaling, inducing platelet aggregate stabilization and triggering a variety of functions important to platelet physiology. This review focuses on the role of beta3 tyrosine phosphorylation in alphaIIbbeta3 outside-in signaling. Tyrosine phosphorylation of beta3 in platelets is a dynamic process which is initiated upon platelet aggregation and also by adhesion of platelets to immobilized fibrinogen. Tyrosine phosphorylation occurs on the beta3 integrin cytoplasmic tyrosine (ICY) domain, a conserved motif found in the beta subunits of several integrins. Beta3 ICY domain tyrosine phosphorylation induces the recruitment of two proteins to the cytoplasmic domains of alphaIIbbeta3: the cytoskeletal protein myosin, important to clot retraction; and the signaling adapter protein Shc, important to platelet stimulation, The critical role of beta3 tyrosine phosphorylation to platelet function was established by the diYF mouse, a novel strain which expresses an alphaIIbbeta3 in which the two beta3 ICY domain tyrosines have been mutated to phenylalanine. These mice are selectively impaired in outside-in alphaIIbbeta3 signaling, with defective aggregation and clot retraction responses in vitro, and an in vivo bleeding defect which is characterized by a pronounced tendency to rebleed. Taken together, the data suggest that the beta3 tyrosine phosphorylation signaling mechanism is important to alphaIIbbeta3 function and might be applicable to a wide variety of integrin mediated events. PMID- 11487014 TI - All in the family: primary megakaryocytes for studies of platelet alphaIIbbeta3 signaling. AB - Integrin alphaIIbbeta3 mediates key platelet adhesive responses during hemostasis and thrombosis. Adhesive ligand binding to alphaIIbbeta3 is regulated by "inside out" signals, while adhesion-dependent cytoskeletal events are regulated by "outside-in" signals from alphaIIbbeta3. Currently, the molecular basis of bidirectional alphaIIbbeta3 signaling is incompletely understood. The functional assessment of integrin signaling pathways in nucleated cells has been facilitated by techniques such as viral transduction which enable expression of dominant active and dominant-inhibitory gene products. This approach cannot be used with anucleate platelets. However, recent advances in the ability to expand human and murine megakaryocytes from hematopoietic stem cells provide a tractable and genetically manipulatable system for studies of alphaIIbbeta3 signaling. This overview will discuss some of the advantages and limitations of this approach and provide examples of its utility. Thus, in addition to their intrinsic value for understanding hematopoiesis and platelet formation, primary megakaryocytes represent a model system complementary to platelets for unraveling the remaining mysteries of alphaIIbbeta3 signaling. PMID- 11487015 TI - Unraveling the mysteries of phospholipid scrambling. AB - Plasma membrane phospholipid asymmetry is maintained by an aminophospholipid translocase that transports phosphatidylserine (PS) and phosphatidylethanolamine (PE) from outer to inner membrane leaflet. Cell activation or injury leads to redistribution of all major lipid classes within the plasma membrane, resulting in surface exposure of PS and PE. Cell surface-exposed PS can serve as receptor sites for coagulation enzyme complexes, and contributes to cell clearance by the reticuloendothelial system. The mechanism(s) by which this PL "scrambling" occurs is poorly understood. A protein called phospholipid scramblase (PLSCR1) has been cloned that exhibits Ca2+-activated PL scrambling activity in vitro. PLSCR1 belongs to a new family of proteins with no apparent homology to other known proteins. PLSCR1 is palmitoylated and contains a potential protein kinase C phosphorylation site. It further contains multiple PxxP and PPxY motifs, representing potential binding motifs for SH3 and WW domains implicated in mediating protein-protein interactions. Although at least two proteins have been shown to associate with PLSCR1, the functional significance of such interaction remains to be elucidated. Evidence that PLSCR1 may serve functions other than its proposed activity as PL scramblase is also presented. PMID- 11487016 TI - The role of ITAM- and ITIM-coupled receptors in platelet activation by collagen. AB - The major activation-inducing collagen receptor glycoprotein VI (GPVI) has been cloned within the last two years. It is a member of the Ig superfamily of proteins and is constitutively associated with the ITAM-bearing Fc receptor gamma chain (FcR gamma-chain). GPVI signals through a pathway that involves several of the proteins used by Fc, B- and T-lymphocyte receptors and which takes place in glycolipid-enriched membrane domains in the plasma membrane known as GEMs. Responses to GPVI are regulated by PECAM-1 (CD31) and possibly other ITIM-bearing receptors. Despite a pivotal role for GPVI, there are important differences between signalling events to collagen and GPVI-specific ligands. This may reflect a role for co-receptors in the response to collagen. PMID- 11487017 TI - The emerging role of the bone marrow-derived stem cells in (therapeutic) angiogenesis. AB - Proper formation of blood vessels (angiogenesis) is essential for development, reproduction and wound healing. When derailed, angionenesis contributes to numerous lifethreatening disorders. While research has generally been focusing on the two main vascular cell types (endothelial and smooth muscle cells), recent evidence indicates that bone marrow may also contribute to this process, both in the embryo and the adult. Novel vascular progenitors, even one common to both endothelial and smooth muscle cells, have been identified in the embryo. An exciting observation is that endothelial precursors have also been identified in the adult bone marrow. Transplantation studies revealed that these precursors as well as other bone marrow-derived cells contribute to the growth of endothelium lined vessels (angiogenesis) as well as the expansion of pre-existing collaterals (arteriogenesis) in ischemic disease. These findings have raised hopes that bone marrow-derived cells might one day become useful for cell-based angiogenic therapy. PMID- 11487018 TI - Protease-activated receptors in vascular biology. AB - G protein-coupled protease-activated receptors (PARs) provide one answer to the question of how coagulation factors and other proteases regulate cellular behaviors. In concert with the coagulation cascade, these receptors provide an elegant mechanism that links mechanical information in the form of tissue injury or vascular leak to cellular responses. Roles for PARs in hemostasis and thrombosis, inflammation, and perhaps even blood vessel development are beginning to emerge. Our current understanding of the role of PARs in platelet and endothelial cell activation and their potential importance in normal and disease states is discussed. PMID- 11487019 TI - Interendothelial junctions and their role in the control of angiogenesis, vascular permeability and leukocyte transmigration. AB - Endothelial cell-cell junctions play an important role in vascular hemostasis. The two junctional proteins VE-cadherin and JAM-1 are localized at adherens and tight junctions, respectively. VE-cadherin is only expressed by endothelial cells, suggesting that it can exert cell specific function. Absence of VE cadherin or blocking of its adhesive activity prevents a normal organization of new vascular structures, suggesting that VE-cadherin may be a molecular target of antiangiogenic therapy. In addition, the ability of permeability-increasing agents and adherent leukocytes to modify VE-cadherin/catenin organization may be related to a role in the control of vascular permeability and leukocyte infiltration. JAM-1 is an integral membrane protein expressed in endothelial and epithelial cells. Its extracellular domain can dimerize and bind homophilically. The intracellular domain (and in particular a PDZ-binding motif) enables JAM-1 to interact with structural and signaling proteins. Study of the molecular interactions of JAM-1 may help explain mechanisms of JAM-mediated function, such as control of paracellular permeability and leukocyte transmigration. PMID- 11487020 TI - Integrin activation. AB - Integrins are cell surface adhesion receptors that participate in a variety of important processes throughout the vasculature. Here we summarize some recent findings on the regulation of integrin mediated cellular adhesion. Particular emphasis is placed on the regulation of integrin affinity for ligand (activation), although this is just one mechanism by which regulation of integrin dependent cell adhesion can occur. Also discussed are recent observations on the structural basis of integrin activation, the role of the cytoplasmic domain in integrin affinity regulation, and potential mechanisms by which activation signals are propagated from integrin cytoplasmic domains to the extracellular ligand binding domain. PMID- 11487021 TI - Plasmin and matrix metalloproteinases in vascular remodeling. AB - Vascular remodeling, defined as lasting structural changes in the vessel wall in response to hemodynamic stimuli, plays a role in many (patho)physiological processes requiring cell migration and degradation of extracellular matrix (ECM). Two proteolytic systems, the fibrinolytic (plasminogen/plasmin) and matrix metalloproteinase (MMP) systems can degrade most ECM components. The availability of mice models with deficiency of main components of both systems has allowed to study their contribution to vascular remodeling in several biological processes. In mouse models of atherosclerosis, urokinase-mediated plasmin generation plays a role in activation of several macrophage-derived MMPs (MMP-3, -9, -12 and -13), triggering elastolysis and collagenolysis, resulting in media destruction and aneurysm formation. Neointima formation after vascular injury, a process that depends on smooth muscle cell migration, is reduced in mice with plasminogen or urokinase deficiency and enhanced in mice with deficiency of TIMP-1 (type 1 tissue inhibitor of MMPs). Also in allograft transplant arteriosclerosis and in abdominal aortic aneurysm both proteolytic systems contribute to matrix degradation. In a mouse model of myocardial infarction, urokinase deficiency protects totally and MMP-9 deficiency partially against cardiac rupture, but these animals suffer cardiac failure. Thus, the plasminogen/plasmin and MMP systems, in concert, contribute to vascular remodeling in the setting of cardiovascular disease. PMID- 11487022 TI - Tissue factor--a receptor involved in the control of cellular properties, including angiogenesis. AB - Tissue factor (TF), the major initiator of blood coagulation, serves as a regulator of angiogenesis, tumor growth and metastasis. In several models, TF expression mediates upregulation of the proangiogenic vasular endothelial growth factor (VEGF) that can directly act on endothelial cells to promote vessel formation. This occurs through ligand binding, activation of signaling cascades, signal transduction and alteration of growth factor expression and is mediated by both, coagulation-dependent and -independent pathways. Depending on the cell type and the biological settings, TF seems to affect cellular properties through (i) factor VIIa (FVIIa)-dependent proteolysis of factor Xa (FXa) and thrombin and subsequent activation of proteinase activated receptor (PAR) -1 and PAR-2, (ii) through direct FVIIa signaling and mitogen activated protein (MAP) kinase activation, that is conferred by a not yet identified receptor, (iii) through interaction of FVII(a) proteolytic activity and signaling of the cytoplasmic domain and (iv) through cytoplasmic signaling independent of ligand binding. The role of phosphorylation of the cytoplasmic domain and the pathways controlling phosphorylation of TF remain poorly understood. PMID- 11487023 TI - AlphaIIbbeta3 and its antagonism at the new millennium. AB - Because of its major role in regulating platelet functions and its prominence on the cell surface, integrin alphaIIbbeta3 has been the subject of intensive investigations. Such studies have provided substantial insights into its structure-function relationships and have led to the development of anti thrombotic drugs that target the receptor. Nevertheless, recent findings have indicated that our understanding of the structure and function of alphaIIbbeta3 remains inadequate. This article addresses two aspects of still evolving alphaIIbbeta3 function: 1) the interface between alphaIIbbeta3 and the blood coagulation system, resulting from interaction of prothrombin with the receptor; and 2) the molecular basis for recognition of the RGD and the fibrinogen gamma chain peptide ligands by alphaIIbbeta3. As illustrated by these two examples, there is still much to be learned about alphaIIbbeta3 if we are to fully appreciate its functions and its potential as a therapeutic target. PMID- 11487024 TI - Extracellular proteolysis and angiogenesis. AB - Extracellular proteolysis is an absolute requirement for new blood vessel formation, a process known as angiogenesis. This review will examine the role of the matrix metalloproteinase and plasminogen activator/plasmin systems during angiogenesis. Extracellular proteolysis has also been implicated in the generation of molecules with angioregulatory activity. These include, but are not limited to, angiostatin and endostatin. However, despite an abundance of data on their bioactivity, the molecular mechanisms by which these molecules achieve their effects are unknown. Anti-proteolysis, particularly in the context of angiogenesis, has become a key target in therapeutic strategies aimed at inhibiting tumor growth and other diseases associated with neovascularization. PMID- 11487025 TI - Atherosclerosis and thrombosis: lessons from animal models. AB - Atherothrombosis defines the occurrence of thrombosis on atherosclerotic lesions. Atherosclerosis is the most prevalent disease of our time and its thrombotic complications are responsible for an exceedingly high number of deaths and disabilities. Over the past few years, experimental investigation and clinical and pathologic observations have led to a better understanding of how a thrombus forms and also of its incidence in acute ischemic syndromes. A thrombus is usually found secondary to atherosclerotic plaque disruption. Mural thrombosis, also at the site of plaque rupture, is an important mechanism in the progression of atherosclerosis even when symptoms are absent. Because atherosclerosis is a silent and asymptomatic disease until complications arise with thrombosis producing clinical symptoms, it is necessary to have models that reproduce the human disease in its early stages. Unfortunately, not all the experimental models of vascular disease have human resemblance and validity. Knowledge of the disease process and of what an experimental animal model can offer is a milestone for a successful investigation. Experimental models of vascular disease have enhanced our understanding of the pathophysiological processes leading to vascular obstruction in both spontaneous and accelerated atherosclerosis and thrombosis. Animal models have provided insight into the role of platelets, lipids, renin angiotensin system (RAS), cytokines and growth factors in the evolution and progression of atherosclerosis and have suggested potential therapeutic interventions. Significant advances in our understanding of the vascular biology and pathology of atherosclerosis and thrombosis, and of the interactions of blood cells, lipids and proteins with the vascular wall, have allowed us to formulate new experimental hypotheses and to test therapeutic strategies, either pharmacological or surgical. PMID- 11487026 TI - Hemostatic risk factors for atherothrombotic disease: an epidemiologic view. AB - A number of prospective epidemiologic studies have now tested the hypothesis that plasma measures of hemostatic factors measured in initially healthy subjects may predict the future onset of atherothrombotic events. Epidemiologic evidence is very strong that plasma fibrinogen is a risk marker, if not a causal factor, for atherothrombotic events. Evidence is mixed on whether factor VII, factor VIII, von Willebrand factor, or other coagulation markers predict incident atherothrombosis. D-dimer is emerging as a strong risk marker, as are several fibrinolytic markers such as tPA antigen--but not PAI-1. Despite suggestive studies, no genetic polymorphism influencing hemostatic factor levels has emerged as a consistent and strong risk factor for atherothrombosis. Thus, prospective epidemiologic studies have identified several potential hemostatic markers of atherothrombosis risk, but their causal role and clinical utility in risk prediction remain incompletely established. PMID- 11487027 TI - Haemostatic genetic risk factors in arterial thrombosis. AB - Haemostasis plays an integral role in arterial thrombotic disease. However, establishing which of the factors are risk factors has proven surprisingly difficult. Because of its technical simplicity and digital nature, the study of haemostatic polymorphisms as risk factors has grown in popularity. Once established as a risk factor, a genetic polymorphism has the potential to aid selective prophylaxis and therapy of disease. Numerous reports have now been published on polymorphisms of coagulation and fibrinolytic factors, of coagulation and fibrinolytic inhibitory proteins, and of platelet membrane glycoprotein receptors. This article describes the polymorphisms and evaluates the results of these studies using the premises of consistency of within-report genotype/phenotype/disease relationships and consistency of outcome between studies. Many studies have been only of association between polymorphisms and disease, a type of study that is prone to error. Furthermore, the collective outcome of these studies has primarily been inconsistent. It is concluded that despite the early promise of polymorphisms as risk factors, fresh approaches differing in scale and design are now required to clarify their possible importance. PMID- 11487028 TI - Plasma lipoproteins, hemostasis and thrombosis. AB - Regulation of hemostasis and thrombosis involves numerous plasma factors that contribute to procoagulant and anticoagulant pathways. Lipid-containing surfaces provide sites where both procoagulant and anticoagulant enzymes, cofactors and substrates are assembled to express their activities. Plasma and lipoproteins can contribute to either procoagulant or anticoagulant reactions. Procoagulant lipids/lipoproteins include triglyceride-rich particles in plasma and oxidized low density lipoprotein (LDL) which can accelerate activation of prothrombin, factor X and factor VII. Potentially anticoagulant lipids and lipoproteins, each of which enhances inactivation of factor Va by activated protein C, include phosphatidylethanolamine, cardiolipin, the neutral glycosphingolipids glucosylceramide and Gb3 ceramide (CD77), and high density lipoprotein (HDL). Remarkably, treatment of hyperlipidemia with statins not only lowers lipids but also provides antithrombotic effects whose mechanisms remain to be clarified. We hypothesize that procoagulant and anticoagulant lipids and lipoproteins in plasma may contribute to a Yin-Yang balance that helps influence the up-regulation and down-regulation of thrombin generation. PMID- 11487029 TI - Risk factors in venous thromboembolism. AB - Venous thromboembolism is a serious disorder because of its potential complications, such as pulmonary embolism and the post-thrombotic syndrome. Inherited determinants of venous thromboembolism are only in part known, but in the past decades considerable progress has been made in the understanding of risk factors for the disease and their clinical impact. In particular, the development of molecular biology techniques and the increasing interest in their application, allowed an identification of two causes of inherited thrombophilia, i.e., factor V Leiden and the prothrombin G20210A mutation. Their recent discovery provided a new approach for improving the knowledge of inherited thrombophilia. In contrast to deficiencies of the naturally occurring anticoagulant proteins antithrombin, protein C and protein S, these two mutations cannot be considered true genetic defects, since they are nucleotide substitutions resulting in a more efficient coagulation process. Since they are rather common in the general populations of Caucasian descent and are associated with a moderate increased risk of venous thromboembolism, the effect of the interaction between inherited and environmental risk factors for venous thromboembolism has become an even greater field of interest. Prevention of first events and recurrences of venous thromboembolism can be optimized only through a knowledge of the main risk factors, their effect, and their interaction with environmental factors. PMID- 11487030 TI - Gene expression in atherogenesis. AB - It is conceivable that the extent and spatio-temperal expression of dozens or even a few hundred genes are significantly altered during the development and progression of atherosclerosis as compared to normal circumstances. Differential gene expression in vascular cells and in blood cells, due to gene-gene and gene environment interactions can be considered the molecular basis for this disease. To comprehend the coherence of the complex genetic response to systemic and local atherosclerotic challenges, one needs accessible high through-put technologies to analyze a panel of differentially expressed genes and to describe the interactions between and among their gene products. Fortunately, new technologies have been developed which allow a complete inventory of differential gene expression, i.e. DD/RT-PCR, SAGE and DNA micro-array. The initial data on the application of these technologies in cardiovascular research are now being reported. This review summarizes a number of key observations. Special attention is paid to a few central transcription factors which are differentially expressed in endothelial cells, smooth muscle cells or monocytes/ macrophages. Recent data on the role of nuclear factor-kappaB (NF-kappaB) and peroxisome proliferation activating receptors (PPARs) are discussed. Like the PPARs, the NGFI-B subfamily of orphan receptors (TR3, MINOR and NOT) also belongs to the steroid/thryroid hormone receptor superfamily of transcription factors. We report that this subfamily is specifically induced in a sub-population of neointimal smooth muscle cells. Furthermore, intriguing new data implicating the Sp/XKLF family of transcription factors in cell-cell communication and maintenance of the atherogenic phenotype are mentioned. A member of the Sp/XKLF family, the shear stress-regulated lung Kruppel-like factor (LKLF) is speculated to be instrumental for the communication between endothelial cells and smooth muscle cells. Taken together, the expectation is that the fundamental knowledge obtained on atherogenesis and the data that will be acquired during the coming decade with the new, powerful high through-put methodologies will lead to novel modalities to treat patients suffering from cardiovascular disease. In view of the phenotypic changes of vascular and blood-borne cells during atherogenesis, therapeutic interventions likely will focus on reversal of an acquired phenotype by gene therapy approach or by using specific drugs which interfere with aberrant gene expression. PMID- 11487031 TI - Vascular-bed-specific hemostasis and hypercoagulable states: clinical utility of activation peptide assays in predicting thrombotic events in different clinical populations. AB - This chapter summarizes the new paradigm for arterial thrombosis. This new paradigm emphasizes the heterogeneity of endothelial cells and the signaling pathways that control endothelial cell gene expression in surrounding tissue. It is suggested that genetic alterations in the signaling pathways are probably responsible for localized thrombosis, as manifested by heart attacks and strokes. A discussion of two clinical studies supporting the new arterial thrombosis paradigm is also included in this chapter. These studies, carried out by genetic engineering in mice, employ activation peptides to help predict the occurrence of thrombotic events in humans. PMID- 11487032 TI - Annexins: key regulators of haemostasis, thrombosis, and apoptosis. AB - Annexins form a family of proteins on the basis of structural and functional criteria. Their physiological relevance is still not fully understood, but is thought to involve phospholipid membrane associated processes. Recent data shed clues about their function in the regulation of thrombosis, haemostasis and apoptosis. These data arise from a multitude of physicochemical, cell biological, animal and clinical studies. This review presents this data and discusses them in the light of the novel understanding that apoptotic cells can contribute to thrombotic and haemostatic processes. PMID- 11487033 TI - Apoptosis as a determinant of atherothrombosis. AB - Clinical manifestations of atherosclerosis are the consequences of atherosclerotic plaque rupture that triggers thrombus formation. Tissue factor (TF) is a key element in the initiation of the coagulation cascade and is crucial in thrombus formation following plaque disruption. TF activity is highly dependent on the presence of phosphatidylserine (PS), an anionic phospholipid that is redistributed on the cell surface during apoptotic death conferring a potent procoagulant activity to the apoptotic cell. Apoptosis occurs in the human atherosclerotic plaque and shed membrane apoptotic microparticles rich in PS are produced in considerable amounts within the lipid core. These microparticles carry almost all TF activity and are responsible for the procoagulant activity of the plaque. Moreover, luminal endothelial cell apoptosis might be responsible for thrombus formation on eroded plaques without rupture. Apoptosis might also play a major role in blood thrombogenicity via circulating procoagulant microparticles that are found at high levels in patients with acute coronary syndromes. PMID- 11487034 TI - Anti-GPIIb/IIIa drugs: current strategies and future directions. AB - Three platelet glycoprotein (GP) IIb/IIIa receptor antagonists have been approved as adjunctive therapy to decrease the ischemic complications of percutaneous coronary interventions (PCI) and/or unstable angina. They include the chimeric murine/human monoclonal antibody 7E3 Fab fragment (abciximab), a cyclic heptapeptide based on the KGD amino acid sequence (eptifibatide), and a nonpeptide mimetic of the RGD sequence (tirofiban). The agents are very effective in providing both short-term and long-term benefit after PCI, and one agent has also demonstrated a progressive long-term mortality benefit. The long-term mortality benefit is highly cost-effective when compared to other medical interventions. The benefits in treating unstable angina without PCI are less dramatic and robust, with some agents providing no benefit. Severe thrombocytopenia is an infrequent, but potentially serious, complication of therapy with all of the agents. The risk of major bleeding is increased only minimally or not at all by the drugs. Currently, a number of new indications for GPIIb/IIIa antagonists are under study, including acute myocardial infarction (+/ thrombolytic therapy, +/- PCI) and stroke. In addition to their impact on improving outcome, the results of clinical trials with these agents provide crucial insights into the contribution of GPIIb/IIIa-mediated platelet function in the pathophysiology of thrombotic vascular disease. PMID- 11487035 TI - Thrombolysis in pulmonary embolism: a debatable indication. AB - Successfully utilized contemporary pulmonary embolism thrombolysis reverses right heart failure rapidly and safely. This therapeutic approach may lower mortality from pulmonary embolism and reduce morbidity from chronic pulmonary hypertension. Pulmonary embolism thrombolysis remains a debatable indication because large clinical trials using survival as an endpoint have not been carried out. Instead, thrombolysis trials have been undertaken with surrogate endpoints such as reduction in clot burden, reduction in pulmonary arterial pressure, and improvement in right ventricular function. In an era where hundreds of thousands of myocardial infarction patients have participated in thrombolysis trials that focus on survival as the principal endpoint, the much smaller trials of PE thrombolysis have not been sufficiently definitive to achieve a consensus. Pharmaceutical companies have not considered this area of investigation to be a good return on investment, because PE is a much less common problem than acute coronary syndromes. No government funding agency has targeted PE thrombolysis as a priority for clinical research. Currently, the only contemporary thrombolytic regimen for pulmonary embolism that is approved by the Food and Drug Administration is tissue plasminogen activator, in a dose of 100 mg/2 h. New thrombolytic agents under development for pulmonary embolism include reteplase, saruplase, and recombinant staphylokinase. Future clinical trials will require multicenter collaboration and focus on clinically relevant endpoints such as reduction of mortality and recurrent venous thromboembolism. PMID- 11487036 TI - The epidemiology of venous thromboembolism in the community. AB - The incidence of venous thromboembolism exceeds 1 per 1000; over 200,000 new cases occur in the United States annually. Of these, 30% die within 30 days; one fifth suffer sudden death due to pulmonary embolism. Despite improved prophylaxis, the incidence of venous thromboembolism has been constant since 1980. Independent risk factors for venous thromboembolism include increasing age, male gender, surgery, trauma, hospital or nursing home confinement, malignancy, neurologic disease with extremity paresis, central venous catheter/transvenous pacemaker, prior superficial vein thrombosis, and varicose veins; among women, risk factors include pregnancy, oral contraceptives, and hormone replacement therapy. About 30% of surviving cases develop recurrent venous thromboembolism within ten years. Independent predictors for recurrence include increasing age, obesity, malignant neoplasm, and extremity paresis. About 28% of cases develop venous stasis syndrome within 20 years. To reduce venous thromboembolism incidence, improve survival, and prevent recurrence and complications, patients with these characteristics should receive appropriate prophylaxis. PMID- 11487037 TI - Thromboembolism in newborns, infants and children. AB - As in adults, acquired and inherited prothrombotic risk factors increase the risk of thrombosis in neonates, infants and children. Duplex sonography, venography, computed tomography and magnetic resonance imaging can be used to diagnose childhood thromboembolism, but venography is the recommended method to confirm vascular occlusion of the upper venous system. After suffering thrombosis, patients should be screened for factor V G1691A, prothrombin G20210A and MTHFR C677T genotypes, deficiencies of protein C, protein S, and antithrombin, elevation of lipoprotein (a) and fasting homocysteine concentrations (3 to 6 months after thrombotic onset: plasma-based assays). Data interpretation is based on age-dependent reference ranges and the identification of causative gene mutations/polymorphisms with respect to the individual ethnic background. Paediatric treatment protocols for acute thromboembolism, including thrombolytic and anticoagulant therapy, are hampered by the lack of appropriate clinical trials. Thus, recommendations from small-scale studies in paediatric patients and guidelines adapted from adult patient protocols may be helpful in the management of paediatric patients on an individual patient basis. PMID- 11487038 TI - Cost-effective diagnosis of deep vein thrombosis and pulmonary embolism. AB - Suspected acute venous thromboembolism is a frequent and challenging clinical problem. Phlebography and pulmonary angiography are costly and invasive and, hence, ill-suited for diagnosing a disease present in only 20% of patients in whom it is suspected. Novel noninvasive instruments, such as plasma D-dimer measurement, lower limb compression ultrasonography and helical CT scan are important breakthroughs in the management of patients with suspected venous thromboembolism. However, none of these instruments is ideal, and they must be combined in rational and cost-effective diagnostic algorithms including clinical assessment, which is increasingly standardized. Such strategies must be validated in management studies, in which patients without venous thromboembolism are not treated and followed up during 3 months. Suspected massive pulmonary embolism is a distinct clinical situation requiring a specific diagnostic approach, in which echocardiography plays a major role. This paper reviews the performance of clinical evaluation and diagnostic tests for venous thromboembolism, and recently validated diagnostic schemes. PMID- 11487039 TI - Heparins and venous thromboembolism: current practice and future directions. AB - Unfractionated heparin (UFH) in adjusted doses and low-molecular-weight heparins (LMWH) in fixed doses are the chosen therapy for the initial treatment of venous thromboembolism. The use of UFH protocols ensures that virtually all patients will promptly achieve the therapeutic range for the activated partial thromboplastin time. However, proper use of UFH requires considerable expertise, can cause inconvenience and has limitations. Unmonitored therapy with subcutaneous LMWH is at least as effective and safe as adjusted-dose UFH, is associated with a considerable reduction of mortality in cancer patients, and permits the treatment of suitable patients in an outpatient setting. LMWH in high prophylactic doses is more effective than UFH and oral anticoagulants for prevention of postoperative venous thrombosis in major orthopedic surgery. Whether thromboprophylaxis should be continued for a few additional weeks after hospital discharge is controversial. LMWH and UFH are equally effective for prevention of postoperative deep-vein thrombosis in cancer patients. In a recent controlled randomized trial, enoxaparin in high prophylactic doses was an effective and safe measure of thromboprophylaxis in ordinary bedridden patients. The efficacy and safety of pentasaccharide (the smallest antithrombin binding sequence of heparin) in the treatment and prevention of venous thromboembolic disorders is currently under investigation. PMID- 11487040 TI - Thrombolysis in deep vein thrombosis: is there still an indication? AB - The most accepted therapy for DVT consists of anticoagulation with unfractionated heparin or low molecular weight heparin, followed by variable duration oral anticoagulation but thrombolytic therapy has been proposed in addition to standard anticoagulation. This paper reviews the literature on post thrombotic syndrome, the natural history of vein patency after therapy, and we perform a systematic review, using accepted standards for meta-analysis, to determine the outcomes when thrombolytic therapy is used to treat DVT. We demonstrate that thrombolytic therapy for DVT results in a significant increase in the risk of major hemorrhage and a significant increase in the rate of vein patency. However, although thrombolytic therapy is advantageous over anticoagulation as measured by early vein patency, a benefit in terms of a reduction in PTS risk, is unproven. Our review also shows that there is no evidence that there is a difference in efficacy between thrombolytic agents or that local therapy differs from systemic therapy. Finally, the potential role of catheter directed therapy is unknown since appropriate trials have not been performed but it is reasonable to use catheter directed therapy in patients with phlegmasia cerulea dolens. We conclude that more work is needed to define the role of thrombolytic therapy but it is too early to abandon this therapeutic modality. PMID- 11487041 TI - Role of coagulation inhibitors in inflammation. AB - It is becoming increasingly clear that coagulation augments inflammation and that anticoagulants, particularly natural anticoagulants, can limit the coagulation induced increases in the inflammatory response. The latter control mechanisms appear to involve not only the inhibition of the coagulation proteases, but interactions with the cells that either generate anti-inflammatory substances, such as prostacyclin, or limit cell activation. Recent studies have demonstrated a variety of mechanisms by which coagulation, particularly the generation of thrombin, factor Xa and the tissue factor-factor VIIa complex, can augment acute inflammatory responses. Many of these responses are due to the activation of one or more of the protease activated receptors. Activation of these receptors on endothelium can lead to the expression of adhesion molecules and platelet activating factor, thereby facilitating leukocyte activation. Therefore, anticoagulants that inhibit any of these factors would be expected to dampen the inflammatory response. The three major natural anticoagulant mechanisms seem to exert a further inhibition of these processes by impacting cellular responses. Antithrombin has been shown in vitro to increase prostacyclin responses and activated protein C has been shown to inhibit a variety of cellular responses including endotoxin induced calcium fluxes in monocytes and the nuclear translocation of NFKB, a key step in the generation of the inflammatory response. In some, but not all, in vivo models, these natural anticoagulants have been able to inhibit endotoxin/E. coli-mediated leukocyte activation and to diminish cytokine elaboration (TNF, IL-6 and IL-8). Phase III clinical studies for treatment of patients with severe sepsis have been completed for APC, which was successful (1), and for antithrombin, which was not (2). A phase III trial with tissue factor pathway inhibitor is in progress. In this review, the mechanisms by which the different natural anticoagulants are thought to function will be reviewed. PMID- 11487042 TI - Physiopathology and regulation of factor XIII. AB - Factor XIII is a plasma transglutaminase. Transglutaminases are at least 8 enzymes which cross-link a number of proteins. This type of reaction not only enhances the original functions of substrate proteins, but also adds new functions to them. Factor XIII in plasma is a tetramer (A2B2), and the A subunit contains the active site. Although transglutaminases are homologous, the nucleotide sequences in their 5'-flanking region differ significantly. Accordingly, transcription factors play a major role in the cell type-specific expression of each transglutaminase. A variety of missense and nonsense mutations, and deletions/insertions with or without out-of-frame shift/premature termination and splicing abnormalities have been identified in the genes for A and B subunits in factor XIII deficiency. In some cases, the mRNA level of the A or B subunit was severely reduced. Molecular and cellular bases have also been explored by expression experiments and by molecular modeling. In most cases, impaired folding and/or conformational change of the mutant A or B subunit leads to both intra- and extra-cellular instability, which is responsible for factor XIII deficiency. PMID- 11487043 TI - Tissue factor: an enzyme cofactor and a true receptor. AB - Tissue factor is considered to be the physiologic trigger of the blood clotting system in normal hemostasis and in many--perhaps most--thrombotic diseases. A wealth of new knowledge is available regarding the structure and assembly of the TF:VIIa complex and the role of factor VIIa and tissue factor in hypercoagulable states. The exciting recent finding that tissue factor can function as a signaling receptor, and suggestions that tissue factor may have important, non hemostatic roles, will be the subject of much additional study in the near future. PMID- 11487044 TI - Roles of platelets and factor XI in the initiation of blood coagulation by thrombin. AB - To account for the variable hemostatic defect in patients with factor XI (FXI) deficiency, with normal hemostasis in contact factor deficiencies, a coagulation paradigm is presented whereby trace quantities of thrombin, generated transiently by exposure of tissue factor at sites of vascular injury, activates FXI bound to the platelet surface in the presence of prothrombin or high Mr kininogen (HK). Tissue factor pathway inhibitor (TFPI) limits the flux of thrombin generated by the tissue factor pathway, and protease nexin II (PNII), released from activated platelets, inhibits solution phase FXIa and localizes FIX activation to the platelet surface where FXIa is protected from inactivation by PNII. Either prothrombin or HK binds to the Apple 1 (A1) domain of FXI, thereby exposing a platelet-binding site in the FXI A3 domain. Dimeric FXI binds to activated platelets directly through the A3 domain of one monomer. After proteolytic activation of platelet-bound FXI by thrombin (or FXIIa), a substrate binding site for FIX is exposed in the opposite monomer that promotes FIX activation on the platelet surface resulting in the local explosive generation of thrombin and the formation of hemostatic thrombi at sites of vascular injury. PMID- 11487045 TI - Protein Z-dependent regulation of coagulation. AB - Protein Z (PZ) is a 62 kDa vitamin K-dependent plasma protein that serves as a cofactor for the inhibition of factor Xa by protein Z-dependent protease inhibitor (ZPI). ZPI is a recently identified 72 kDa member of the serpin superfamily of proteinase inhibitors that contains a tyrosine at its reactive center. PZ circulates in plasma in a complex with ZPI. Inhibition of factor Xa by ZPI in the presence of phospholipids and Ca++ is enhanced 1000-fold by PZ, but ZPI also inhibits factor XIa in a process that does not require PZ, phospholipids or Ca++. ZPI activity is consumed during coagulation through proteolysis mediated by factor Xa with PZ and factor Xla. Concomitant PZ deficiency dramatically increases the severity of the prothrombotic phenotype of factor VLeiden mice. Studies to determine the potential roles of PZ and ZPI deficiency in human thrombosis are in progress. PMID- 11487046 TI - Antiphospholipid syndrome: diagnostic aspects of lupus anticoagulants. AB - Antiphospholipid syndrome (APS) is an autoimmune disorder in which antiphospholipid antibodies (aPL) are thought to be involved in the development of venous and/or arterial thrombosis. APL found in this syndrome are antibodies directed against a variety of phospholipid (PL) binding-proteins of which beta3 glycoprotein I (beta2GPI) and prothrombin are considered to be the major antigens. Some of these antibodies prolong PL-dependent clotting reactions and are termed lupus anticoagulants (LA). Autoimmune aPL which bind through beta2GPI to cardiolipin are called anticardiolipin antibodies (aCL). Clinical studies indicate that LA is a stronger risk factor for thrombosis than aCL. The production of monoclonal antibodies against beta2GPI and prothrombin has enabled us to understand the mechanism by which LA prolong coagulation in vitro. LA form bivalent antigen-antibody complexes with increased affinity for PL which compete with coagulation factors for the same catalytic surface. These LA positive monoclonal antibodies may be helpful in further improving the laboratory diagnosis of LA. PMID- 11487047 TI - Genetic approach to thrombophilia. AB - Venous thrombosis is a multifactorial disease. Multiple interactions between genetic and environmental factors contribute to the development of the disease. Presently, we know of six or seven genetic risk factors for venous thrombosis. However, together these defects can explain the clustering of thrombotic events in only a small subset of families with thrombophilia. As to the identification of new genetic risk factors for thrombosis, we seem to have arrived at the end of a practicable road with the classical approach of thrombophilia, which usually starts with the study of the association of hemostatic phenotypes and thrombotic risk. At the same time we have undertaken various genetic approaches aiming at identifying polymorphisms/mutations causing thrombotic risk. This review summarizes what we have learnt so far, what to do and what not to do. The odds for finding remaining common genetic risk factors for venous thrombosis during the next ten years may be predicted to be fairly high. PMID- 11487048 TI - Cell survival and clinical outcome following intrastriatal transplantation in Parkinson disease. AB - Intrastriatal transplantation of embryonic dopaminergic neurons is currently explored as a restorative cell therapy for Parkinson disease (PD). Clinical results have varied, probably due to differences in transplantation methodology and patient selection. In this review, we assess clinical trials and autopsy findings in grafted PD patients and suggest that a minimum number of surviving dopaminergic neurons is required for a favorable outcome. Restoration of [18F] fluorodopa uptake in the putamen to about 50% of the normal mean seems necessary for moderate to marked clinical benefit to occur. Some studies indicate that this may require mesencephalic tissue from 3-5 human embryos implanted into each hemisphere. The volume, density and pattern of fiber outgrowth and reinnervation, as well as functional integration and dopamine release. are postulated as additional important factors for an optimal clinical outcome. For neural transplantation to become a feasible therapeutic alternative in PD, graft survival must be increased and the need for multiple donors of human embryonic tissue substantially decreased or alternate sources of donor tissue developed. Donor cells derived from alternative sources should demonstrate features comparable to those associated with successful implantation of human embryonic tissue before clinical trials are considered. PMID- 11487049 TI - Plaque-induced abnormalities in neurite geometry in transgenic models of Alzheimer disease: implications for neural system disruption. AB - Neurites that pass through amyloid-beta deposits in Alzheimer disease (AD) undergo 3 changes: they develop phosphorylated tau immunoreactivity; the density of SMI-32-positive dendrites diminishes; and they also develop a marked alteration in their geometric features, changing from being nearly straight to being quite curvy. The extent to which the latter 2 phenomena are related to phosphorylated tau is unknown. We have now examined whether amyloid-beta deposits in APP695Sw transgenic mice, which have only rare phosphorylated tau containing neurites. develop these changes. We found that dendritic density is diminished within the boundaries of amyloid-beta plaques, with the greatest loss (about 80%, p < 0.001) within the boundaries of thioflavine S cores. Remaining dendrites within plaques develop substantial morphological alterations quantitatively similar to those seen in AD. A statistically significant but smaller degree of change in geometry was seen in the immediate vicinity around plaques, suggesting a propagation of cytoskeletal disruption from the center of the plaque outward. We examined the possible physiological consequences of this change in dendritic geometry using a standard cable-theory model. We found a predicted delay of several milliseconds in about one quarter of the dendrites passing through a thioflavine S plaque. These results are consistent with previous observations in AD, and suggest that thioflavine S-positive amyloid-beta deposits have a marked effect on dendritic microarchitecture in the cortex, even in the relative absence of phosphorylated tau alterations. PMID- 11487050 TI - Oxidative damage is the earliest event in Alzheimer disease. AB - Recently, we demonstrated a significant increase of an oxidized nucleoside derived from RNA, 8-hydroxyguanosine (8OHG), and an oxidized amino acid, nitrotyrosine in vulnerable neurons of patients with Alzheimer disease (AD). To determine whether oxidative damage is an early- or end-stage event in the process of neurodegeneration in AD, we investigated the relationship between neuronal 8OHG and nitrotyrosine and histological and clinical variables, i.e. amyloid-beta (A beta) plaques and neurofibrillary tangles (NFT), as well as duration of dementia and apolipoprotein E (ApoE) genotype. Our findings show that oxidative damage is quantitatively greatest early in the disease and reduces with disease progression. Surprisingly, we found that increases in A beta deposition are associated with decreased oxidative damage. These relationships are more significant in ApoE epsilon4 carriers. Moreover, neurons with NFT show a 40%-56% decrease in relative 8OHG levels compared with neurons free of NFT. Our observations indicate that increased oxidative damage is an early event in AD that decreases with disease progression and lesion formation. These findings suggest that AD is associated with compensatory changes that reduce damage from reactive oxygen. PMID- 11487051 TI - Experimentally induced leptomeningeal glioneuronal heterotopia and underlying cortical dysplasia of the lateral limbic area in rats treated transplacentally with methylmercury. AB - Leptomeningeal glioneuronal heterotopia (LGH) is a developmental anomaly sometimes observed at the surface of human brains with severe malformations. We experimentally induced LGH in brains of rat pups by transplacental exposure to methylmercury. Histopathological profiles of the induced LGH, including the spatio-temporal predominance of the manifestation, suggest some aspects of the histogenesis of this malformation. Pregnant rats on embryonic day 8 (E8), E11, E13, E16, E18 or E21 were treated orally with a single administration of 20 mg/kg methylmercury chloride, and the brains of their delivered offspring were examined on postnatal day 7 (P7) and P28. The incidence of LOH varied significantly according to the treatment day: it was almost exclusively restricted to individuals treated on E13. Furthermore, all the induced LGH was confined to the subarachnoid space dorsal to the rhinal fissure, unilaterally or bilaterally. A part of the nest was connected to the underlying cortical surface of the lateral limbic area, where glia limitans and basal lamina were disrupted. Narrow stripes of disarrangement of cortical neurons underlying the bridges were observed. The P7 LGHs consisted mainly of neurons, some of which were GABA-immunolabeled. and a small number of astrocytes as well as endogenous blood vessels and fibroblasts. LGHs in P28 brains consisted mainly of GFAP-positive astrocyte processes. An additional experiment with double administrations of 5-bromo-2-deoxyuridine (BrdU) and methylmercury on E13 disclosed an abnormally widespread distribution of labeled neurons throughout all cortical layers underlying the LGH in P28 brains. Thus, cerebral LGH would be induced if a developing brain were insulted at the early stage of neurogenesis. accompanied by cortical dysplasia. PMID- 11487052 TI - Cholesterol accumulates in senile plaques of Alzheimer disease patients and in transgenic APP(SW) mice. AB - Mounting evidence suggests that cholesterol may contribute to the pathogenesis of Alzheimer disease (AD). We examined whether cholesterol might be present in senile plaques, a hallmark neuropathological feature of AD. We employed 2 different fluorometric-staining techniques (filipin staining and an enzymatic technique) for the determination of cholesterol in brains of postmortem confirmed AD patients and in nondemented, age-matched histopathologically normal controls. AD patient brains showed abnormal accumulation of cholesterol in congophilic/birefringent dense cores of senile plaques that was essentially absent in histopathologically normal controls. To determine whether increased senile plaque-associated cholesterol occurred generally in all plaques or was restricted to a specific subset, quantitative analysis was performed. Data indicate abnormal accumulation of cholesterol in cores of mature plaques but not in diffuse or immature plaques. Additionally, transgenic mice that overexpress the "Swedish" amyloid precursor protein (Tg APP(SW), line 2576) exhibited a similar pattern of abnormal cholesterol accumulation in mature, congophilic amyloid plaques at 24 months of age that was absent in their control littermates or in 8-month-old Tg APP(SW) mice (an age prior to amyloid deposition). Taken together, our results imply a link between cholesterol and AD pathogenesis and suggest that cholesterol plays an important role in the formation and/or progression of senile plaques. PMID- 11487053 TI - Long-term cortical atrophy after excitotoxic striatal lesion: effects of intrastriatal fetal-striatum grafts and implications for Huntington disease. AB - It is not currently clear whether the cortical atrophy observed in Huntington disease (HD) is entirely a direct consequence of the disease or at least partially a secondary consequence of striatal atrophy. This is of major importance for evaluating the possible therapeutic value of intrastriatal fetal striatum grafts in HD. Cresyl violet-stained sections from rats that had received striatal excitotoxic lesions 1 wk or 4 wk previously showed small and statistically nonsignificant decreases in the thickness of cortical layers V and VI, while series from rats lesioned 12 months previously showed marked decreases in the thickness of the whole cortex (approximately 35% decrease), layer V (approximately 45%-50%) and layer VI (approximately 45%-50%), together with marked neuron loss in these layers. In deep layer V and layer VI, Fluoro-Jade staining showed labeled neurons in animals lesioned 1 wk previously, labeled neurons and astrocytes in animals lesioned 4 wk previously, and practically no labeling in animals lesioned 12 months previously. Intracortical injection of Phaseolus vulgaris leucoagglutinin revealed that corticostriatal fibers were practically absent from the lesioned area of striata lesioned 12 months previously. However, rats that received intrastriatal fetal-striatum grafts shortly after the lesion and were killed 12 months later showed a significant reduction in cortical atrophy, and a large number of labeled corticostriatal fibers surrounding and innervating the graft. In addition, a reduction in the number of Fluoro-Jade-labeled cells in the cortex was already apparent at 3 wk post-grafting. Regardless of whether HD has a primary effect on the cortex, the present results suggest that the striatal degeneration caused by HD contributes markedly to the cortical atrophy, and that intrastriatal grafts may ameliorate this secondary component of the cortical degeneration. PMID- 11487054 TI - Inhibition of antigen-specific T cell trafficking into the central nervous system via blocking PECAM1/CD31 molecule. AB - Trafficking of antigen-specific T cells into the central nervous system (CNS) is an important initiating step in inflammation in the brain. In spite of the extensive knowledge about the role of adhesion molecules in T cell migration across peripheral vessels, the mechanism of the entry of antigen-specific T cells into the CNS is not known. This work was designed to study the regulatory roles of adhesion molecules in antigen-specific T cell migration into the CNS. Antigen specific T cells were tracked in an in vivo migration assay using T cell receptor (TCR) transgenic mice having 95% of T cells specific for a defined antigen. pigeon cytochrome c (PCC). TCR transgenic mice were cannulated intraventricularly (IVT) for PCC antigen infusion and cerebrospinal fluid (CSF) sampling. Upon PCC infusion into the CNS, the number of alpha/beta TCR+ Vbeta3+ Mac1- cells in the CSF was characterized in the presence or absence of anti-adhesion molecule reagents. We found that antibodies against VCAM-1 (CD106), VLA-4 (CD49d/CD29), ICAM-1 (CD54), and LFA-1 (CD11a/CD18) did not influence the increased number of antigen-specific T cells in the CSF However, upon intravenous (i.v.) injection, anti-PECAM-1 (CD31) antibody or PECAM-Ig chimeric molecule inhibited the trafficking of alpha/beta TCR+ Vbeta3+ Mac1- cells into the CNS. The expression of PECAM-1 (CD31) was also up-regulated on antigen-specific T cells in a time dependent manner in vitro upon antigenic stimulation. The antigen-induced activation of T cells in vivo was measured by CD44 and LFA-1 expression and found to be comparable between mPECAMIg-treated mice and wild-type serum control treated groups. This indicates that CD31 inhibition of antigen-specific T cell accumulation in the CNS is probably not due to a functional inhibition of these cells. Finally, adoptive transfer of CFSE-labeled AND transgenic cells into naive animals resulted in the accumulation of these cells in the CNS upon PCC IVT immunization that was also inhibited by mPECAMIg treatment. Hence, PECAM-1 (CD31) might play an important role in regulating antigen-specific T cells trafficking in CNS inflammatory diseases. PMID- 11487055 TI - Concurrent hypermethylation of multiple genes is associated with grade of oligodendroglial tumors. AB - Current evidence suggests that epigenetic changes play an important role in the evolution of human cancers. In this study, we evaluated whether hypermethylation of CpG islands at the gene promotor regions of several tumor-related genes is involved in the carcinogenesis of oligodendroglial tumors. We examined the methylation status of 11 genes in a series of 43 oligodendroglial tumors (19 oligodendrogliomas, 13 anaplastic oligodendrogliomas, 9 oligoastrocytomas, and 2 anaplastic oligoastrocytomas) by methylation-specific polymerase chain reaction. Our results showed that hypermethylation of CpG islands was detectable in 8 of 11 genes studied and 74% of tumors were hypermethylated in at least 1 gene. Promotor hypermethylations were detected in O6-methylguanine-DNA methyltransferase (MGMT), RB1, estrogen receptor, p73, p16INK4a, death-associated protein kinase, p15INK4b, and p14ARF at 60%, 34%, 30%, 16%, 12%, 10%, 7%, and 2%, respectively. No hypermethylation was detected in the promotors of glutathione-S-transferase P1, von Hippel-Lindau or the DNA mismatch repair (hMLH1) genes. Statistical analysis revealed that concordant hypermethylation of at least 2 genes, p16INK4a and p15INK4b were significantly associated with anaplastic oligodendroglial tumors, and hypermethylation of MGMT was significantly associated with loss of chromosome 19q and with combined loss of chromosomes 1p and 19q. More importantly, several candidate tumor suppressor genes such as p16INK4a, p15INK4b, and p73 that were previously reported as unmutated in oligodendroglial tumors were found to be hypermethylated in their CpG islands. Taken together, we conclude that hypermethylation of CpG islands is a common epigenetic event that is associated with the development of oligodendroglial tumors. PMID- 11487056 TI - Histopathology, electrodiagnostic testing, and magnetic resonance imaging show significant peripheral and central nervous system myelin abnormalities in the cat model of alpha-mannosidosis. AB - Alpha-mannosidosis is a disease caused by the deficient activity of alpha mannosidase, a lysosomal hydrolase involved in the degradation of glycoproteins. The disease is characterized by the accumulation of mannose-rich oligosaccharides within lysosomes. The purpose of this study was to characterize the peripheral nervous system (PNS) and central nervous system (CNS) myelin abnormalities in cats from a breeding colony with a uniform mutation in the gene encoding alpha mannosidase. Three affected cats and 3 normal cats from 2 litters were examined weekly from 4 to 18 wk of age. Progressively worsening neurological signs developed in affected cats that included tremors, loss of balance, and nystagmus. In the PNS, affected cats showed slow motor nerve conduction velocity and increased F-wave latency. Single nerve fiber teasing revealed significant demyelination/remyelination in affected cats. Mean G-ratios of nerves showed a significant increase in affected cats compared to normal cats. Magnetic resonance imaging of the CNS revealed diffuse white matter signal abnormalities throughout the brain of affected cats. Quantitative magnetization transfer imaging showed a 8%-16% decrease in the magnetization transfer ratio in brain white matter of affected cats compared to normal cats, consistent with myelin abnormalities. Histology confirmed myelin loss throughout the cerebrum and cerebellum. Thus, histology, electrodiagnostic testing, and magnetic resonance imaging identified significant myelination abnormalities in both the PNS and CNS that have not been described previously in alpha-mannosidosis. PMID- 11487057 TI - Screening of Pleurotus ostreatus isolates for their ligninolytic properties during cultivation on natural substrates. AB - Thirteen basidiospore-derived isolates of Pleurotus ostreatus f6 strain differing in the level of ligninolytic enzyme production and other characteristics (mycelium extension rate, colony morphology) from the parental strain were cultivated on natural substrates. Under these conditions ligninolytic enzyme activity, loss of organic mass, polycyclic aromatic hydrocarbons (PAHs) degradation and colonization of sterile and nonsterile soil were studied. The activity of ligninolytic enzymes was substantially higher in straw than in liquid culture, although the differences between the isolates were less pronounced on this substrate. Some of the isolates showed a very good ability to decompose the lignocellulosic substrate (straw) and a relatively high loss of organic mass was found after 50 days of cultivation in these strains. The original strain f6 and isolates B13 and B26 successfully degraded all seven tested PAH compounds present in experimental soil samples, but the higher or lower ligninolytic enzyme production of isolates tested had no substantial effect on the extent of the degradation. In our screening, six basidiospore-derived isolates growing well in nonsterile soil were found, which could be suitable for the prospective biotechnological exploitation. PMID- 11487058 TI - Biodegradation and conversion of alkanes and crude oil by a marine Rhodococcus sp. AB - A hydrocarbon degrader isolated from a chronically oil-polluted marine site was identified as Rhodococcus sp. on the basis of morphology, fatty acid methyl ester pattern, cell wall analysis, biochemical tests and G + C content of DNA. It degraded up to 50% of the aliphatic fraction of Assam crude oil, in seawater supplemented with 35 mM nitrogen as urea and 0.1 mM phosphorus as dipotassium hydrogen orthophosphate, after 72 h at 30 degrees C and 150 revolutions per minute. The relative percentage of intracellular fatty acid was higher in hydrocarbon-grown cells compared to fructose-grown cells. The fatty acids C16, C16:1, C18 and C18:1 were constitutively present regardless of the growth substrate. In addition to these constitutive acids, other intracellular fatty acids varied in correlation to the hydrocarbon chain length supplied as a substrate. When grown on odd carbon number alkanes, the isolate released only monocarboxylic acids into the growth medium. On even carbon number alkanes only dicarboxylic acids were produced. PMID- 11487059 TI - Use of hydrophobic membranes to supply hydrogen to sulphate reducing bioreactors. AB - This paper reports on the application of hydrophobic membranes to supply the gaseous substrates hydrogen/carbon dioxide (H2/CO2) to a sulphate reducing bioreactor. For this, two flat 0.016 m2 sheets of flouroplast microporous (0.45 microm) membranes were inserted in a 3.6 dm3 bioreactor for the supply of H2/CO2 gas as small gas bubbles. The bioreactor was operated at 30 degrees C and pH 7.0 and was also equipped with an external ultra filtration module for biomass retention. At a sulphate loading rate (SLR) of 1.32 g SO4(2-) dm(-3) day(-1) and a hydraulic retention time (HRT) of 61 h, a sulphate reduction rate (SRR) of 0.90 g SO4(2-) dm(-3) day(-1) was achieved. When the influent sulphate concentration was reduced from 3.36 to 0.75 g SO4(2-) dm(-3) by lowering the HRT to 10.3 h (SLR of 1.75 g SO4(2-) dm(-3) day(-1) the SRR dropped to 0.22 g SO4(2-) dm(-3) day( 1). The lower sulphate reduction efficiency was most probably caused by a too short biomass-substrate contact time or by irreversible sulphide inhibition. Mass transfer limitation of H2 and improper mixing of the reactor liquid were shown not to contribute to the low sulphate reduction efficiency. PMID- 11487060 TI - Pyrene is metabolized to bound residues by Penicillium janthinellum SFU403. AB - We have previously shown that the filamentous fungus, Penicillium janthinellum SFU403 (SFU403) oxidizes pyrene to pyrene 1,6- and 1,8-quinones and that the level of pyrenequinones (PQs) subsequently declines suggesting that PQs are not terminal metabolites. The purpose of this study was to determine the fate of PQs in SFU403. First, we compared the fate of 14C-pyrene in SFU403 and a non-pyrene oxidizing fungus, a Paecilomyces sp.. After 7 days of incubation, more than 80% of the radioactivity was cell-associated in both fungi; however, while 90% of the 14C could be extracted from the Paecilomyces sp. as unmetabolized pyrene, 65-80% of the bound radioactivity remained inextractable from SFU403. Further evidence that pyrene oxidation to PQs was required for irreversible binding was obtained by comparing the extent of 14C bound to SFU403 when it was grown for 21 days under conditions that resulted in differing amounts of 14C-pyrene oxidation. The results showed that approximately 40% of the inextractable products were bound residues derived from pyrene metabolites. The balance (60%) could be attributed to strong sorption of unreacted pyrene. We used electron paramagnetic resonance spectroscopy and oxygen consumption studies to demonstrate that both NADPH and glutathione can reduce PQs by one electron to their corresponding semiquinone anion radicals in vitro. These studies demonstrate that PQs are metabolized by SFU403 to bound residues, possibly via semiquinone intermediates. PMID- 11487061 TI - Quinones as terminal electron acceptors for anaerobic microbial oxidation of phenolic compounds. AB - The capacity of anaerobic granular sludge for oxidizing phenol and p-cresol under anaerobic conditions was studied. Phenol and p-cresol were completely converted to methane when bicarbonate was the only terminal electron acceptor available. When the humic model compound, anthraquinone-2,6-disulfonate, was included as an alternative electron acceptor in the cultures, the oxidation of the phenolic compounds was coupled to the reduction of the model humic compound to its corresponding hydroquinone, anthrahydroquinone-2,6-disulfonate. These results demonstrate for the first time that the anaerobic degradation of phenolic compounds can be coupled to the reduction of quinones as terminal electron acceptor. PMID- 11487062 TI - Degradation of microbial polyester poly(3-hydroxybutyrate) in environmental samples and in culture. AB - Poly(3-hydroxybutyrate) [P(3HB)] test-pieces prepared from the polymer produced by Azotobacter chroococcum were degraded in natural environments like soil, water, compost and sewage sludge incubated under laboratory conditions. Degradation in terms of % weight loss of the polymer was maximum (45%) in sewage sludge after 200 days of incubation at 30 degrees C. The P(3HB)-degrading bacterial cultures (36) isolated from degraded test-pieces showed different degrees of degradation in polymer overlayer method. The extent of P(3HB) degradation increases up to 12 days of incubation and was maximum at 30 degrees C for majority of the cultures. For most efficient cultures the optimum concentration of P(3HB) for degradation was 0.3% (w/v). Supplementation of soluble carbon sources like glucose, fructose and arabinose reduced the degradation while it was almost unaffected with lactose. Though the cultures degraded P(3HB) significantly, they were comparatively less efficient in utilizing copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate [P(3HB-co-3HV)]. PMID- 11487063 TI - Transformation of 2,4,6-trichlorophenol by the white rot fungi Panus tigrinus and Coriolus versicolor. AB - The toxicity of thirteen isomers of mono-, di-, tri- and pentachlorophenols was tested in potato-dextrose agar cultures of the white rot fungi Panus tigrinus and Coriolus versicolor. 2,4,6-Trichlorophenol (2,4,6-TCP) was chosen for further study of its toxicity and transformation in liquid cultures of these fungi. Two schemes of 2,4,6-TCP addition were tested to minimize its toxic effect to fungal cultures: stepwise addition from the moment of inoculation and single addition after five days of growth. In both cases the ligninolytic enzyme systems of both fungi were found to be responsible for 2,4,6-TCP transformation. 2,6-Dichloro-1,4 hydroquinol and 2,6-dichloro-1,4-benzoquinone were found as products of primary oxidation of 2,4,6-TCP by intact fungal cultures and purified ligninolytic enzymes, Mn-peroxidases and laccases of both fungi. However, primary attack of 2,4,6-TCP in P. tigrinus culture was conducted mainly by Mn-peroxidase, while in C. versicolor it was catalyzed predominantly by laccase, suggesting a different mode of regulation of these enzymes in the two fungi. PMID- 11487064 TI - Surfactant inhibition of bacterial growth on solid anthracene. AB - Surfactants have been proposed as a promising method to enhance bioremediation of hydrophobic compounds in contaminated soils. However, the results of effects of surfactants on bioremediation are not consistent. This study showed that Triton X 100 at low concentration (0.024 mM or 0.09 CMC) inhibited the rate of growth of either a Mycobacterium sp. or a Pseudomonas sp. on solid anthracene as sole carbon source. Recovery of microbial growth rate could be achieved by dilution of surfactants, while addition of more surfactant gave an immediate decrease in growth rate. No inhibition of growth by Triton X-100 was observed with growth on glucose. The surfactant sorbed onto the surfaces of both the cells and the anthracene particles, which could inhibit uptake of anthracene. The results were consistent with the hypothesis that inhibition of microbial adhesion of cells to anthracene was responsible for the inhibition of growth by Triton X-100. PMID- 11487065 TI - Increased expression of atrial natriuretic peptide in ureteral obstructed kidneys in rats. AB - OBJECTIVES: Whether the natriuresis in the previously ureteral obstructed kidney may be related to an altered regulation of local atrial natriuretic peptide (ANP) was investigated. MATERIALS AND METHODS: Male Sprague-Dawley rats were unilaterally obstructed of the left ureters for 48 hours. The left and right ureters were separately cannulated to collect urine samples. In some rats, both kidneys were removed without releasing the obstruction. Control rats were treated the same except that no ureteral obstruction was made. The mRNA expression of ANP and natriuretic peptide receptor (NPR)-A was determined in the kidney by reverse transcription-polymerase chain reaction. The guanylyl cyclase activity was measured by the amount of cGMP generated in response to ANP. Plasma levels of ANP were measured by radioimmunoassay. RESULTS: The ureteral obstruction did not significantly affect the plasma ANP levels. In the obstructed kidney, the creatinine clearance was decreased, while the fractional excretion of sodium increased. The expression of ANP mRNA was increased in the obstructed kidney. The NPR-A mRNA expression was not altered in the glomerulus, but was decreased in the papilla of the obstructed kidney. Nor was the guanylyl cyclase activity in the glomerulus altered. Although the guanylyl cyclase activity in the papilla was significantly decreased in the obstructed kidney, it was rapidly resumed upon releasing the obstruction. CONCLUSIONS: An increased local expression of ANP may, at least in part, account for the natriuresis in the previously ureteral obstructed kidney. PMID- 11487066 TI - Impact of nocturnal enuresis on children and young people. AB - An understanding of the psychological impact of nocturnal enuresis has been consistently hampered by inter-changeability of terminology, varied inclusion criteria, diverse methodologies and equivocal findings. This paper reviews the impact on children and young people by examining both qualitative and quantitative findings. Aspects of functioning that are explored include social adjustment, emotional state, personality, self- concept and behaviour. Generally the findings suggest children do experience bedwetting as distressing but those with mono-symptomatic nocturnal enuresis are no different from the normal population or from matched controls across all aspects of functioning. Those children most vulnerable to psychological distress as a consequence of bedwetting are identified. Interestingly there is consistent evidence for improvement in emotional functioning and self-esteem following treatment although this may not be necessarily due to the treatment itself. Methodological issues are addressed in terms of future research directions. PMID- 11487067 TI - Genetic and gender influences on nocturnal bladder control--a study of 2900 3 year-old twin pairs. AB - OBJECTIVE: The present study of over 2900 twin pairs born in England and Wales in 1994 examines the influences of genetics and gender on nocturnal bladder control at 3 years of age. MATERIALS AND METHOD: Parent report data was analysed in terms of means and components of variance, using a sex-limitation model to explore genetic and environmental variation within and between the sexes. RESULTS: Both genetics and gender are seen to influence acquisition: bladder control at 3 years is moderately heritable (24%), and girls show on average slightly increased acquisition compared with boys, even within opposite-sex pairs. The sex limitation modelling showed an interaction between genetic influence and gender whereby nocturnal bladder control was significantly more heritable in boys (33%) than girls (10%). CONCLUSIONS: Both genetics and gender are important and interacting factors in the aetiology of nocturnal bladder control. PMID- 11487068 TI - Primary nocturnal enuresis and left-handedness. AB - To date, no previous studies have described the association between primary nocturnal enuresis and left-handedness. In our series of enuretic patients, the prevalence of left-handedness was significantly higher than the control group. These data suggest a new correlation which needs further investigation. PMID- 11487069 TI - Reconstructive urology in the nordic countries--a hospital questionnaire survey. AB - A hospital survey of adult reconstructive urologic surgery in the Nordic countries is presented. The response rate was 80% and included most general hospitals and university clinics. Despite similarities between the healthcare systems of the various countries several differences were found. Cystectomy was performed in a large number of institutions in all countries except Denmark. The annual number of orthotopic bladder substitutions per institution was calculated as three to four (range of medians for each country) and the number of continent cutaneous diversions as two to seven. Open urethral procedures were performed more frequently in Sweden than in the other countries. Surgery for penile curvature and implantation of three-component prostheses for erectile dysfunction was more commonly performed in Denmark and Iceland compared to Sweden. PMID- 11487070 TI - Clam ileocystoplasty: successful treatment of severe bladder overactivity. AB - OBJECTIVE: When conservative treatment for bladder overactivity fails the "clam" ileocystoplasty has been proposed to restore continence and preserve urethral voiding. This study presents our experience with this technique. MATERIALS AND METHODS: Thirty patients, 4 men and 26 women, with a mean age of 50.3 years (range 21-71 years) were operated upon. Five patients had an incomplete spinal lesion and detrusor hyperreflexia, 25 subjects had an idiopathic, unstable detrusor. The ileocystoplasty was combined with a colposuspension procedure in 8 patients. In one patient simultaneous repair of an urethro-vaginal fistula was performed using omentum flap interposition. The mean follow-up was 60 months (range 4-127). RESULTS: To the surgeon, 27 patients (90%) reported satisfaction with bladder control and relief of symptoms at follow-up. When asked by a nurse in a written questionnaire, 18 out of 23 patients (78%) were quite happy with their bladder function postoperatively. No serious complications to surgery were noted. Postoperatively, 11 patients used self-intermittent catheterization and one, by choice, preferred an indwelling catheter. Due to failure, two patients later underwent reoperation with urinary diversion and one further patient required an additional artificial sphincter. Metabolic consequences owing to enteric malabsorption were infrequent and of a mild nature. No malignant change was identified. CONCLUSION: In patients with refractory overactive bladder, we found the "clam" ileocystoplasty to be an effective technique for symptom reduction and increased quality of life. PMID- 11487071 TI - Validation of a computer version of the patient-administered Danish prostatic symptom score questionnaire. AB - OBJECTIVE: To validate a computer version of the Danish Prostatic Symptom Score (DAN-PSS) questionnaire and compare it with the paper version. MATERIAL AND METHODS: Ninety-three male patients (aged 25-87 years), referred to a department of urology for lower urinary tract symptoms (LUTS), filled in a personal computer (PC) version and a paper version of the DAN-PSS questionnaire. Subsequently they answered a questionnaire concerning their preferences and computer experience. RESULTS: A fair correlation between the total scores from the paper and PC versions was found. The differences were independent of both total score and age. In the PC version all LUTS questions were answered while 9.8% were left blank in the paper version. The sexual questions were answered by 71% of patients in the paper version and by 87.5% in the PC version. For the questions in the PC version a learning curve was observed in terms of the time taken to answer the questions. Previous computer experience did not influence answering time, difference in score between paper and PC versions or total score. Almost all patients preferred the PC version to the paper version. CONCLUSION: The PC scores are reasonably reliable judging by comparison with previously validated traditional paper scores. The PC questionnaire had a higher response rate and was preferred by the majority of patients. PMID- 11487072 TI - Two-year follow-up results of a prospective randomized trial comparing hybrid laser prostatectomy with TURP in the treatment of big benign prostates. AB - OBJECTIVE: We compared conventional transurethral electroresection of the prostate (TURP) and hybrid laser treatment in patients with symptomatic bladder outflow obstruction caused by a benign prostate bigger than 40 ml. MATERIAL AND METHODS: Forty-six patients with symptomatic urodynamically confirmed outflow obstruction caused by benign prostate hyperplasia bigger than 40 ml were accepted to the prospective trial and were randomized to receive hybrid laser treatment or TURP. The hybrid laser technique involved initial non-contact Nd:YAG coagulation followed by contact Nd:YAG vaporization to open the prostatic urethra. Patients were re-assessed after 3, 6, 12 and 24 months. RESULTS: Of the 46 patients, 37 (80%) were available at the 24-month follow-up. The re-operation rate was 14.3% in the hybrid laser group and 8.3% in the TURP group. The decrease in DanPSS-1 symptom score from baseline at 24 months was 51.0% in the hybrid laser group (p<0.01) and 80.0% in the TURP group (p<0.001), with no statistically significant differences between the groups. Early improvement in objective urinary parameters of peak urinary flow rate and residual urinary volume showed deterioration in hybrid laser group during longer follow-up, whereas in TURP group the improvement of these variables was sustained. Comparison between groups showed TURP to be superior in peak urinary flow rate (p < 0.001) and residual urinary volume (p < 0.01) at the 24-month follow-up visit. CONCLUSIONS: Both hybrid laser treatment and TURP give good symptomatic relief lasting at least 2 years in the treatment of big obstructing prostates, but hybrid laser treatment is associated with a higher re-operation rate because of inadequate initial prostate tissue removal and with inferior outcome in objective urinary parameters compared with TURP. PMID- 11487073 TI - Urethral injury secondary to pelvic fracture: anatomical and functional classification. AB - OBJECTIVE: To develop a new functional and anatomical classification of urethral injury secondary to pelvic fracture. MATERIAL AND METHODS: Fifty-six male patients (20 children, 36 adults) with urethral injuries secondary to pelvis fracture were evaluated. Clinical examination, retrograde urethrogram immediately after the accident, operative findings in the emergency state, subsequent combined retrograde urethrography and suprapubic cystography, operative findings during urethral reconstruction and postoperative follow-up were critically reviewed retrospectively. RESULTS: A new classification of urethral injury secondary to pelvic fracture is proposed as a result of our findings: Type 1. Injury to the prostate; 1a. Proximal avulsion of the prostate; 1b. Incomplete or complete trans-prostatic rupture. Type 2. Stretching of the membranous urethra. Type 3. Incomplete or complete pure rupture of the prostatomembranous junction, supradiaphragmatic. Type 4. Incomplete or complete rupture of the bulbomembranous urethra, infradiaphragmatic. Type 5. Variable combined urethral injuries affecting more than one level of the urethra, prostatic and membranous or prostatomembranous and bulbomembranous, injury to proximal sphincteric mechanism combined with prostatic and/or membranous urethral injury. CONCLUSIONS: This anatomical and functional classification includes all types of urethral injuries secondary to pelvic fracture; moreover, it directs the attention towards evaluation of the urethral sphincteric mechanism, which is essential for the therapeutic and medicolegal aspects. PMID- 11487074 TI - Penile incarceration with metal objects--a review of procedure choice based on penile trauma grade. AB - OBJECTIVE: To outline treatment guidelines according to level of penile trauma for penile incarceration by metal devices. METHOD: A post-1950 (hand-held powered cutting tool era) Medline search was performed. Cases were divided into four groups: string techniques and variants with and without aspiration of blood from the glans; aspiration techniques; cutting devices; and surgical techniques. Trauma grade (according to Bhat et al., 1991), site time (incarceration time), removal technique, removal time, anesthesia and recovery time were assessed. RESULTS: The string technique and variants were used for grades 1-3. They had short removal (30-120 min), site (3-72 h) and recovery (1-24 h) times. Occasional glans decompressive with blood aspiration was required. Anesthesias included none (wrapping without glans aspiration), i.m. morphine and general (glans aspiration). Pure aspiration techniques used multiple needle punctures for grades 2-3. Aspiration cases had short site times (8-14 h), but required a spinal or general anesthesia. Cutting device cases (grades 1-5) required general anesthesia, had a short removal times (45-90 min), but long site (7 h-30 days) and recovery (2-66 days) times. Surgical degloving was utilized mainly for grade 5 cases, required spinal or general anesthesia, had short site (2-30 days), but long recovery (9-28 days). CONCLUSIONS: The string, wrapping, aspiration techniques and cutting devices are suited for grades 1-3. Cutting requires a shield to avoid blade trauma and water-cooling to prevent thermal injury. Suspected underlying devitalized tissue (e.g. grade 4) is examined by Wood's lamp. Failure to identify gangrenous tissue will result in post interventional complications and a prolonged recovery time. PMID- 11487075 TI - The impact of internal spermatic artery ligation during laparoscopic varicocelectomy on recurrence rate and short post operative outcome. AB - OBJECTIVE: To prospectively compare the recurrence rate and short postoperative outcome after randomized laparoscopic varix ligation with internal spermatic artery (ISA) preservation versus laparoscopic varix ligation with ISA ligation. MATERIAL AND METHODS: Twenty-five patients with 35 varicocele who required varix ligation for infertility in 13 patients, scrotal pain in 15 patients and scrotal swelling in 2 patients who underwent one of two procedures: laparoscopic varix ligation with ISA prservation (Group A) or laparoscopic varix ligation with ISA ligation (Group B) were postoperatively evaluated for short post operative outcome and underwent percutaneous spermatic venograms to detect recurrence. Fisher's Exact Test was used for statistical analysis. RESULTS: Recurrence through parallel collaterals was noted in 39% and 5.9% in Group A and Group B respectively as demonstrated on percutaneous spermatic venous venography (PSV) (statistically significant p = 0.0408). Preoperative pain completely resolved in all patients in Group B and persisted in 45% in Group A. However, this was not statistically significant (p = 0.088). No testicular atrophy or hydrocele formation was noted in either group. CONCLUSIONS: Laparoscopic varix ligation with ISA ligation has lower recurrence rate than laparoscopic varix ligation without ISA ligation and may provide better varicocele related pain control with no increase in hydrocele or testicular atrophy rate. We recommend ISA ligation routinely during laparoscopic varix ligation. PMID- 11487076 TI - Factors affecting the fate of prolonged forgotten 'J' stents. AB - PURPOSE: To compare the effects of age and presence or absence of renal impairment at the time of 'J' stent insertion on the subsequent fate of prolonged forgotten stents. PATIENTS AND METHODS: A patient was described as having a prolonged forgotten 'J' stent, if the patient's 'J' stent was removed more than 12 months after it had been inserted. We compared the effect of age at insertion and presence or absence of renal impairment on the complications encountered in such patients. We describe briefly the management of the complications. We defined a patient as having moderate renal impairment if the patient has serum creatinine >200 < 500 micromol/l and is not on dialysis. RESULTS: We treated 17 patients with forgotten 'J' stents between 1994 and 2000. Fifteen were adults, mean age 25 (range 18-72) years, and 2 were children 9 and 10 years respectively at the time the stents were inserted. The mean duration of stent retention was 24.30 (range 12-60) months. In 12 patients the stents were forgotten for between 12 and 18 months. In these, the stents had varying degrees of calcification but were easily removed intact endoscopically in 11 out of 12 cases. One 10-year-old boy in this group required open surgical removal of the stent. In one 35-year-old patient, the stent was forgotten for 36 months. It had fractured spontaneously in 7 places and required endoscopic and open removal of stent fragments. In 2 cases, a growing 9-year-old boy, and a 30-year-old man the stents were forgotten for 46 and 48 months respectively. After 46 months of retaining the stent, the stent spontaneously fractured in 11 places in the growing child, while in the adult it became heavily calcified and fractured during attempts to remove it endoscopically. Two adult patients with moderate renal failure at the time of stent insertion retained the stents for 40 and 60 months respectively. One of these 2 stents had a minor calcification at the tip of the stent in the renal pelvis. Both stents were removed intact endoscopically and showed no sign of fracture or calcification. CONCLUSION: In a growing child a prolonged forgotten 'J' stent is very likely to undergo spontaneous fracture due to the stress exerted on it as a result of cranio-caudal growth of the child. In adults, prolonged forgotten stents become calcified, brittle and lose tensile strength after more than one year of placement and may fracture either spontaneously or during attempts to remove them endoscopically. In patients producing hypotonic urine such as patients with moderate renal failure, a prolonged forgotten stent may remain little affected by the passage of time. PMID- 11487077 TI - Mannose-binding lectin gene polymorphism associated with the patterns of glomerular immune deposition in IgA nephropathy. AB - OBJECTIVE: To elucidate the genetic background underlying the diversity of mesangial immune deposition in IgA nephropathy (IgAN), we investigated the distribution of mannose-binding lectin (MBL) gene codon 54 polymorphism and serum MBL levels in IgAN patients. METHODS: Seventy-seven IgAN patients with glomerular IgA and C3 deposits (Group A) and 70 with glomerular IgA, IgG, IgM, C3 and Clq deposits (Group AGM) were included in the present study. Control group consisted of 140 normal adults. MBL genotypes were investigated by polymerase chain reaction and restriction fragment length polymorphism. Serum MBL levels with different genotypes were also assayed in some subjects. RESULTS: The variant allele (GAC) was markedly associated with Group AGM (OR = 1.95, 95% C.I.: 1.06 3.58). In both Group A and Group AGM, more patients carrying the variant allele had episodes of upper respiratory or gastrointestinal infections prior to onset or exacerbation of IgAN than wild homozygotes (GGC/GGC). In addition, a significant difference in serum MBL level was also observed between wild homozygotes and heterozygotes (GGC/GAC) (GGC/GGC > GGC/GAC) (p<0.0001) in all groups, while there was no difference for subjects with the same genotypes among the three groups (p > 0.05). Serum MBL levels of the rare variant homozygotes approached zero. CONCLUSIONS: Our findings provide evidence that the host defense molecule, MBL, may be involved in the formation of the diversity of glomerular immune deposition in IgAN. Genetic deficiency of MBL may partially account for abundant immune deposits in some IgAN patients. PMID- 11487078 TI - The relative prognosis of nodular and diffuse diabetic nephropathy. AB - MATERIAL AND METHODS: One hundred and forty-four diabetic patients with biopsy proven diffuse diabetic glomerulosclerosis (DIF), 134 patients with nodular diabetic nephropathy (NOD) and 152 diabetic patients with nondiabetic-related morphology (104 chronic nephropathy, 48 primary GN) were followed for up to 12 years to determine the clinical prognosis. RESULTS: Comparing the NOD patients with the DIF patients, there were more females (41% vs 26%, p < 0.05) and they were more often uremic at biopsy (24% vs 12%, p < 0.01), but the age was similar (53.3 years vs 50.1 years, NS). There was no difference in diabetes type I and II incidence. Compared with the general population, the odds ratio (OR) for death was 7.2 (confidence interval 5.5-9.5) for DIF and 10.8 (8.5-13.7) for NOD. The OR for combined renal or patient death was: DIF 15.2 (11.7-19.7); NOD 24.6 (19.4 31.0). After correction for age, sex, and pre-existing uremia, NOD had a 1.70 (p < 0.01) times increased risk of death compared with DIF, and a 2.42 (p < 0.01) times increased risk of renal failure. The life expectancy for NOD was 4.0 years, and average time to dialysis was 2.1 years. NOD prognosis was similar to other chronic nephropathy. The incidence of all atherosclerotic complications except AMI was twice as high in NOD than DIF. Diabetes type had no influence on prognosis. The estimated incidence of diabetic nephropathy was 56/mio/year. CONCLUSION: Nodular diabetic nephropathy has a poorer prognosis than diffuse due to a higher rate of atherosclerotic and uremic complications. PMID- 11487079 TI - Angiotensin converting enzyme gene insertion/deletion polymorphism in idiopathic nephrotic syndrome in Kuwaiti Arab children. AB - OBJECTIVE: Angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism influence the circulating and cellular levels of ACE and has been shown to be a risk factor in a number of diseases including IgA nephropathy. We have investigated the association of ACE gene I/D polymorphism with the clinical presentation of idiopathic nephrotic syndrome (INS) in Kuwaiti children. MATERIALS AND METHODS: The genotypes for ACE gene I/D polymorphism were determined in 102 subjects (54 INS cases and 48 healthy controls) using a PCR method. RESULTS: The distribution of DD, ID and II genotypes was 70%, 20% and 10% in INS cases compared with 52%, 46% and 2% in the controls. The mean age of onset of the disease was significantly lower in the INS cases with DD genotype (37 months) compared with cases with II genotype (65 months, p < 0.05). The clinical manifestation of the disease was considerably severe in cases with DD genotypes compared with cases having ID and II genotypes. The INS cases with DD genotype also showed a significantly higher incidence of steroid sensitivity and steroid dependence. Seventy-three per cent of the INS cases with minimal change lesion had a DD genotype. Also 70% of the cases which needed cytotoxic drugs had DD genotype. CONCLUSION: Our data suggest an association of the D-allele of the ACE gene I/D polymorphism with the clinical manifestation of INS in Kuwaiti Arab children. PMID- 11487080 TI - Testicular microlithiasis needs long-term surveillance. AB - We present a case of testicular microlithiasis in which testicular biopsy failed to predict the development of a testicular tumour. PMID- 11487081 TI - Pyeloureteral junction stenosis and ureteral valve causing hydronephrosis. AB - A young female patient underwent ureteral resection and Hynes-Anderson pyeloureteroplasty because of pyeloureteral junction stenosis and a ureteral valve (UV), which caused hydronephrosis. Despite using ultrasonography and intravenous urography the ureteral valve was observed only on exploration. UV should be considered as a possible cause of upper urinary tract obstruction; retrograde urography can be essential. PMID- 11487082 TI - Leiomyosarcoma of the bladder fourteen years after cyclophosphamide therapy for retinoblastoma. AB - We report a case of bladder leiomyosarcoma which occurred in a 22-year-old female who had been treated with cyclophosphamide for a period of 68 months for retinoblastoma diagnosed at 18 months postpartum. Partial cystectomy was performed. Forty-two months after the operation she is tumor-free. PMID- 11487083 TI - Subacute hyponatraemia after transurethral resection of the prostate. AB - We present a case of subacute hyponatraemia which developed 3 days after a transurethral resection of the prostate. Symptoms consisted of nausea, vomiting and headache. Release of vasopressin due to excessive surgical bleeding, combined with liberal oral and intravenous administration of sodium-free fluids, was considered to be the cause. PMID- 11487084 TI - Rupture of an abdominal aortic aneurysm due to aortic wallstent dislocation masquerading as renal colic. AB - Placement of an aortic wallstent for treatment of an abdominal aortic aneurysm (AAA) is a frequent therapeutic measure. Whereas AAA is known to mimic renal colic, aortic wallstent dislocation is a novel diagnostic problem. Herein, we report the first case of a patient with a dislocated aortic wallstent and subsequent aneurysm rupture and discuss appropriate diagnostic measures. PMID- 11487085 TI - An unusual cause of incontinence: urethral coitus. AB - Megalourethra and incontinence due to urethral coitus are very rare phenomena in women. In this paper, we report two cases of urinary incontinence due to urethral coitus where there was no vaginal anomaly or dysfunction. PMID- 11487086 TI - Spontaneous regression of bilateral hydronephrosis due to retroperitoneal fibrosis. AB - The case was a 73-year-old male who presented with bilateral hydronephrosis due to retroperitoneal fibrosis. Later he became anuric and nephrostomy was constructed. However, with no medical treatment, the hydronephrosis disappeared. Such a spontaneous regression is rare and the etiology is discussed. PMID- 11487087 TI - Characteristics of transport of fluoresceinated methotrexate in rat small intestine. AB - The small intestinal damage induced by the methotrexate (MTX) treatment results in malabsorption and diarrhea. The fluoresceinated methotrexate (F-MTX) may possibly be useful to study such effects of MTX on the small intestine. The purpose of this study is to characterize the transport of F-MTX in the small intestine in order to use it as a membrane transport and cellular marker of MTX. The transport of F-MTX in the rat small intestine (jejunum) was examined in the in vitro everted segments of the intestine. The uptake was pH-dependent and showed a maximal effect at pH 6.0, which was the same as the results of MTX previously reported. Further, it was temperature-dependent and was inhibited by metabolic inhibitors, dinitrophenol and sodium azide, and by MTX. The transport kinetics at pH 6.0 in the mucosal solution and at pH 7.4 in the serosal solution was saturable with Km of 0.48 +/- 0.23 microM and Vmax of 0.66 +/- 0.24 pmol/cm/min and in addition, the passive diffusion was observed there. These results suggested that the transport of F-MTX was energy-dependent and was mediated by the same transporter as that of MTX, although, in addition to it, other transport mechanism might contribute to the F-MTX transport. Therefore F MTX will be of great use to investigate the MTX transport system in the normal and diseased states of small intestine, using various fluorescence techniques like visualization of membrane-associated transport proteins. PMID- 11487088 TI - Oleoyl-estrone does not have direct estrogenic effects on rats. AB - The estrogenic effects of oleoyl-estrone (OE) administration, either though continuous i.v. infusion with osmotic minipumps or administered by daily oral gavage, were studied. Binding of OE to human recombinant purified alpha receptors was negligible, and that of estrone (E1) was only a fraction of 17beta-estradiol (E2) binding. Intravenous--but not oral--OE administration resulted in marked increases of both E1 and E2 in rat plasma, but oral OE did not induce significant changes in either plasma hormone in Wistar or Zucker rats. The weight of uteri and ovaries increased with time of administration in Zucker rats treated with i.v. OE, but inguinal mammary gland proliferation between subcutaneous adipose tissue was even more marked. Oral administration of OE, however, did not increase either uterine weight or mammary gland proliferation, even at doses (10 micromol/kg x d) higher than those given i.v. (3.5 micromol/kg x d). The results indicate that i.v. administration of OE resulted in limited estrogenic effects mainly due to the high accumulation of E1 giving rise to significant increases in E2. On the other hand, oral administration of OE, even at higher daily doses, did not increase the circulating levels of either estrogen and, therefore, there were no significant effects on mammary gland proliferation or uterine weight. The oral administration of OE as a slimming drug, then, do not result in estrogenic side effects over a wide range of daily doses. PMID- 11487089 TI - Intestinal handling of an oral oleoyl-estrone gavage by the rat. AB - Adult Zucker lean (Fa/?) female rats received a single 250 nmol oral gavage of 3H labelled oleoylestrone in 0.2 ml of sunflower oil. After one hour, samples of arterial, portal and suprahepatic blood, and lymph were obtained and fractioned to determine the amount of radioactivity present in the form of free estrone, acyl-estrone and hydrophilic estrone esters in the blood of each vessel. Lipoprotein fractions (chylomicra + VLDL, LDL, HDL and lipoprotein-depleted plasma) were also analysed as well as the distribution of absorbed 3H-estrone in the intestine, specific organs and carcass. About one third of the oleoyl-estrone dose recovered was found in the tissues, mainly in the blood, the rest remaining relatively untouched in the intestinal content. High hypothalamic estrone uptake (compared with the rest of the brain) was observed. Data from non-radioactive estrone measurements showed a similar pattern of absorption and tissue distribution to that obtained by 3H-estrone tracking alone. In both cases, most of the estrone present in the intestinal lumen was absorbed as intact oleoyl estrone, but a significant part was absorbed as free estrone. There is a net transfer of 3H-estrone into portal blood HDL, and part of the 3H-estrone is also loaded into lymph-carried chylomicra. A large share of free estrone is filtered by the liver, but most of the acyl-estrone absorbed passes unaltered. The oral administration of oleoyl-estrone results in significant absorption of the unaltered molecule, which is transferred to lymph-carried chylomicra and also directly to plasma HDLs. It may be inferred that the HDL fraction contains the physiological carrier of oleoyl-estrone in its role of ponderostat signal. PMID- 11487090 TI - Withdrawal and bidirectional cross-withdrawal responses in rats treated with adenosine agonists and morphine. AB - The aim of this study was to investigate whether the A1/A2 receptor agonist, 5'-N ethylcarboxamidoadenosine (NECA), and the selective A1 agonist, N6 cyclopentyladenosine (CPA), induced physical dependence by quantifying specific antagonist-precipitated withdrawal syndromes in conscious rats. In addition, the presence of bidirectional cross-withdrawal was also investigated. The agonists were administered s.c. to groups of rats at 12 h intervals. Antagonists were administered s.c., 12 hours after the last dose, followed by observation and measurement of faecal output for 20 min. NECA (4 x 0.03 mg kg(-1), s.c) and CPA (4 x 0.03, 0.1 and 0.3 mg kg(-1), s.c.) induced physical dependence, as shown by the expression of a significant withdrawal syndrome when challenged with the adenosine A1/A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX, 0.1 mg kg(-1), s.c.) and the A1 antagonist, 8-cyclopentyl-1,3-dipropylxanthine (CPDPX, 0.1 mg kg(-1), s.c.) respectively. The syndromes consisted of teeth chattering and shaking behaviours shown to occur in morphine-dependent animals withdrawn with naloxone viz, paw, body and 'wet-dog' shakes, but with the additional behaviours of head shaking and yawning. In further contrast to the opiate withdrawal syndrome, no diarrhoea occurred in the groups of animals treated with adenosine agonists and withdrawn with their respective antagonists. Bidirectional cross-withdrawal syndromes were also revealed when naloxone (3 mg kg(-1), s.c.) was administered to adenosine agonist pre-treated rats and adenosine antagonists were given to morphine pre-treated rats. This study provides further information illustrating that close links exist between the adenosine and opiate systems. PMID- 11487091 TI - Role of LPS and receptor subtypes in the uptake of TNF by the murine lung. AB - Tumor necrosis factor alpha (TNF) is an important mediator in lung injury. The kinetics of TNF uptake by the lung are not completely understood. In this study, we evaluated the role that lipopolysaccharide (LPS) and the two types of TNF receptor (p55 and p75) play in the uptake of circulating murine TNF by the murine lung. TNF radioactively labeled with 125I (I-mTNF) was administered intravenously (2 x 10(6) cpm/mouse) to mice with both receptors (wild-type) or to mice missing one (p55-/- or p75-/-) or both (p55-/- and p75-/-) TNF receptors. Blood to lung non-reversible sequestration (Ki) and reversible uptake (Vi) were measured with multiple-time regression analysis. Uptake by lung of I-mTNF in wild-type mice had reversible and non-reversible components. This uptake was decreased by intratracheal, but not by intravenous, LPS, suggesting modulation by local, rather than systemic, inflammation. The p75-/- deficient mice retained the Ki (saturable, non-reversible) component of TNF uptake, whereas p55-/- deficient mice retained the Vi (saturable, reversible) component of TNF uptake. Both Ki and Vi components of TNF uptake were absent in the lungs of p55-/- p75-/- deficient mice. These studies show that local inflammation inhibits the uptake of circulating I-mTNF by lung and that uptake consists of two distinguishable compartments: reversible uptake mediated by the p75 receptor and non-reversible sequestration mediated by the p55 receptor. PMID- 11487092 TI - Periodic maternal deprivation and lesion of anterodorsal thalami nuclei induce alteration on hypophyso adrenal system activity in adult rats. AB - The hypothalamic-pituitary-adrenal (HPA) axis is normally regulated by extrahypothalamic limbic structures, among these, the anterodorsal thalami nuclei (ADTN), which exert an inhibitory influence on HPA, in basal and acute stress conditions in rats. In the present work we have investigated whether neonatal maternal deprivation (MD) produces long-term changes in the ADTN regulation of HPA activity. Maternal deprivation, in female rats, for 4.5 hs daily, during the first 3 weeks of life, produced at 3 months old, a significant decrease in plasma ACTH concentration (p<0.001) and an increase in plasma corticosterone (C) (p<0.001), compared to control non-deprived rats (NMD). Also MD showed higher plasma epinephrine (E) and norepinephrine (NE) levels than NMD rats. The increase of NE (66.6% p<0.001) was higher than that observed in E (19%). After 30 days of ADTN lesion, plasma ACTH values were higher than in sham lesioned rats, in both NMD and MD animals. ACTH response was greater in MD rats. Plasma C, in NMD, was higher, whereas in MD lesioned animals, it was significantly lower than in sham lesioned. In MD rats, lesion produced a significant increase in plasma E and NE (p<0.001), and again, NE increase was higher than E increase. The more accentuated increase of NE than E, suggests sympathetic nervous system hyperactivity. In summary, neonatal maternal deprivation induces long-term alterations on HPA axis sensitivity and medullo adrenal secretion; enhanced sympathetic nervous system activity and, therefore affected the ADTN inhibitory influence on ACTH and adrenal glands secretion. PMID- 11487093 TI - Progesterone and 17 beta-estradiol acutely stimulate nitric oxide synthase activity in rat aorta and inhibit platelet aggregation. AB - The rapid non-genomic stimulatory action of progesterone (Pg) and estradiol (E2) on nitric oxide synthase (NOS) activity of endothelium intact aortic rings and its effect on platelet aggregation was investigated. First we measured the effect of the hormones on platelet aggregation when added to rat aortic strips (RAS) incubated in a PRP. RAS induced an antiaggregatory activity, which was enhanced by the presence of the hormones. The inhibitory action induced by the hormones was evoked in a dose dependent manner (10 pM-100 nM). These effects are specific for progesterone and 17-beta-estradiol, since either testosterone and 17-alpha estradiol were devoid of activity. The hormones induced rapid responses, producing significant inhibition within 1 to 5 minutes of hormonal exposure. The addition of 10(-5) M L-NAME suppressed the antiaggregatory effect of 1 nM E2 or 10 nM Pg. Furthermore, we specifically quantified the NO generation by the 3H citrulline technique. 10(-8) M E2 induced 2-fold increase of RAS citrulline production, while the increment induced by 10(-7) M Pg was 55% over control. Preincubation with 10(-5) M L-NAME completely suppressed the stimulatory action of 10(-9) M E2 or 10(-8) M Pg, confirming that the antiaggregatory factor released from the aortic tissue was NO. Preincubation with cycloheximide did not block the increment in NO induced by the hormones. In conclusion the present study provides for the first time evidence of acute, non-genomic effects of Pg on rat aorta NOS activity and platelet aggregation in coincidence with the results obtained with estradiol treatment. PMID- 11487094 TI - Digitalis-like factor response to hyperinsulinemia accompanying a euglycemic hyperinsulinemic clamp or oral glucose tolerance test. AB - Many studies of essential hypertension find evidence of insulin resistance in the same individuals, leading some to postulate a hypertensive role for insulin. However, the mechanisms by which insulin might exert a hypertensive effect are not fully resolved. An endogenous sodium pump inhibitor or digitalis-like factor (DLF) has been proposed as a hypertensive agent and its plasma concentrations are elevated in hypertension and in Type II diabetes, where insulin levels are elevated. Hence, we studied the effect of insulin on DLF using two approaches to achieve hyperinsulinemia. Normotensive men and women underwent a hyperinsulinemic, euglycemic clamp (40 mU/m2/min insulin, 40 mU = 1.6 x 10(-6) g) in which plasma insulin concentration was kept at high, but physiologic levels. Serum DLF (measured as inhibition of [Na,K]ATPase activity) and insulin levels were measured at baseline and every 30 min throughout the 2 hr clamp. Additionally, other subjects underwent an oral glucose tolerance test (OGTT) as a second means of increasing insulin levels. Insulin and DLF levels were measured prior to and hourly for 3 hours after receiving 100 gm of oral glucose. Serum DLF increased significantly during the clamp from a baseline of 4.6 +/- 0.81 to a peak of 8.7 +/- 1.2% inhibition (p=0.001). Comparison of the baseline and peak DLF levels with concomitant plasma insulin levels revealed a significant correlation (R=0.60, p=0.003). During the OGTT, DLF levels rose from a baseline of 2.4 +/- 1.0 to a peak level of 5.0 +/- 0.4%, p = 0.04. These results suggest that DLF, a factor that can cause vascular smooth muscle contraction and potentially influence blood pressure, is increased by hyperinsulinemia and provides a mechanism by which insulin may increase blood pressure. PMID- 11487095 TI - Specific site methylation in the 5'-flanking region of CYP1A2 interindividual differences in human livers. AB - Human cytochrome P4501A2 (CYP1A2) is involved in the metabolism of a large number of common drugs and is responsible for the metabolic activation of numerous promutagens and procarcinogens. Large interindividual differences exist in the expression of this enzyme. This variability has important implications for drug efficacy and cancer susceptibility. In this sudy, the methylation status of the CCGG site (bp -2759) located adjacent to an AP-1 site in the 5'-flanking region of the CYP1A2 gene was assessed in liver samples from a pool of 55 human donors. DNA methylation is an important epigenetic mechanism controlling gene expression and may be one of the molecular mechanisms underlying the interindividual variation. Analysis was conducted using Hpa II digestion and PCR. Results showed that individual samples varied in the methylation status at this site. The site was found to be hypermethylated in approximately 60% of the samples. To compare methylation status with level of CYP1A2 expression, results were analyzed by median test. In 44% of the samples with expression levels above the median the CCGG site was hypermethylated. However, for those samples with levels below the median hypermethylation of the site was found in 73% of the samples. The difference was statistically significant (p<0.05). These findings indicate that CpG methylation may be involved in controlling the expression of CYP1A2, with hypermethylation reducing expression. Moreover, this approach can be useful in assessing the role of site-specific DNA methylation in interindividual variation of CYP1A2. Analysis of other CpG sites in potentially important regulatory elements of the CYP1A2 gene will continue. PMID- 11487096 TI - Preference for corn oil in olfactory-blocked mice in the conditioned place preference test and the two-bottle choice test. AB - We studied the effects of olfactory stimuli on preference for corn oil in mice. In the conditioned place preference test, voluntary intake of 100% corn oil by both olfactory normal and ZnSO4-induced olfactory-blocked (anosmic) mice resulted in their place preference for the corn oil-related box. In the olfactory normal mice, place preference was also observed by voluntary intake of linoleic acid as well as of corn oil. In the two-bottle choice test, normal mice showed significant preference for test fluids that contained corn oil at all concentrations (1-10%) tested relative to vehicle alone. However, the lower concentrations (1 and 3%) of corn oil were not preferred in the anosmic mice. These results suggested that stimuli other than olfaction contributed to the rewarding effects of corn oil, but at lower concentrations olfactory stimuli might act as a signal for the oil. PMID- 11487097 TI - The hypnotic effect of propofol in the medial preoptic area of the rat. AB - Recent introduction of the intravenous anesthetic propofol as an ICU sedative has allowed a deeply sedated state to be maintained for extended periods in the ICU without delays in emergence. Although such sedation has been advocated to promote physiologic sleep, little evidence exists to support such a strategy. To explore propofol's effect on sleep regulation, we administered propofol directly into the medial preoptic area (MPA) of the rat, an anatomic site where administration of other sedatives (triazolam and phenobarbital) also induce sleep. We performed three two-hour sleep studies in the daytime with the lights on following the administration of propofol (8 ng or 40 ng) or vehicle (intralipid). The higher dose of propofol significantly reduced sleep latency and increased nonREM and total sleep times when compared to vehicle. REM sleep times, intermittent waking times and number of transitions were not altered. Mean nonREM sleep bout length was increased significantly at the higher dose. These findings suggest that propofol may enhance sleep by acting at a hypothalamic site. PMID- 11487098 TI - Effects of impurities on oxygen transfer rates in diffused aeration systems. AB - A series of unsteady-state reaeration tests were performed in a 500-L tank at 0.81-4.58 m3/h diffused-air flow rate and 288-302 K water temperature. Three different types of impurities: soybean oil, surfactant, and diatomaceous earth were doped to simulate the impurities in wastewaters and the effects of the impurities on the oxygen transfer rate were investigated. The ASCE and the two zone oxygen mass-transfer models were used to analyze the unsteady-state reaeration data and the volumetric mass-transfer coefficients determined from the unsteady-state reaeration data were correlated as a function of the diffused-air flow rate, water temperature, and impurity concentration. The results showed that the alpha factors based on the ASCE model are less sensitive to the impurity concentration while the presence of the impurities significantly reduces the alpha factors in the gas bubble zone. The saturation DO concentration and volumetric oxygen mass-transfer rate can be predicted by the two-zone model along with the correlation obtained in this study. PMID- 11487099 TI - Determination of cyanobacterial hepatotoxins directly in water using a protein phosphatase inhibition assay. AB - A colorimetric phosphatase inhibition assay using protein phosphatase 2A and p nitrophenyl phosphate as substrate for determining cyanobacterial peptide hepatotoxins directly in water without sample preconcentration has been developed. The assay uses commercially available materials and is much more simple to use than similar procedures using radiolabelled substrates. It has similar sensitivity to the radiolabelled assays and, with a working range of around 0.2-1 microg/L, is able to determine these toxins at concentrations below the provisional World Health Organisation guideline of 1 microg/L for microcystin LR. The method appears robust and not to be affected by the sample matrix apart from possibly some components of cellular material if present at very high levels in extracts of cyanobacterial material. It is not affected by the presence of low levels of methanol in sample extracts. PMID- 11487100 TI - Estimation of dissolved organic carbon (DOC) concentrations in nanoliter samples using UV spectroscopy. AB - A simple analytical system has been developed and tested for estimating dissolved organic carbon (DOC) concentrations in nanoliter samples of sediment porewaters. The system consists of a loop injector that introduces 200 nL of sample directly into the capillary tubing connected to a scanning UV-VIS detector equipped with a 35-nanoliter capillary flowcell. Analysis of DOC by spectroscopic and high temperature catalytic oxidation (HTCO) methods on the same samples showed a strong linear correlation between UV absorbance at 254 nm and HTCO-DOC concentration, allowing the DOC quantification. The simplicity and the robustness of the UV system allow DOC measurements in the field at a rate of 30 samples/h. We have used the UV system successfully for determining high-resolution distributions of porewater DOC concentrations in sediments at millimeter scales. PMID- 11487101 TI - Fate of plasticised PVC products under landfill conditions: a laboratory-scale landfill simulation reactor study. AB - The long-term behaviour of plasticised PVC products was investigated in laboratory-scale landfill simulation reactors. The examined products included a cable material and a flooring with different combinations of plasticisers. The objective of the study was to assess whether a degradation of the PVC polymer or a loss of plasticisers occurred under landfill conditions. A degradation of the polymer matrix was not observed. The contents of plasticisers in aged samples was determined and compared to the respective original products. The behaviour of the various plasticisers was found to differ significantly. Losses of DEHP and BBP from the flooring were too low for analytical quantification. No loss of DIDP from the cable was detectable, whereas DINA in the same product showed considerable losses of up to 70% compared to the original contents. These deficits were attributable to biodegradation rather than leaching. There was no equivalent release of plasticisers into the leachate. PMID- 11487102 TI - Microbial reduction of perchlorate in pure and mixed culture packed-bed bioreactors. AB - Perchlorate (ClO4-) has been detected in a large number of surface and ground waters in the US. Due to health concerns of perchlorate in drinking water, the California Department of Health Services has established a provisional action level of 18 microg/L. Several microbial isolates have been obtained capable of microbiological perchlorate reduction through cell respiration, but few of these have been tested for perchlorate removals to these low levels. The feasibility of using one isolate (KJ) for water treatment was tested in a packed-bed bioreactor by comparing minimum detention times necessary to achieve complete removal of perchlorate. Perchlorate was reduced approximately from 20 mg/L to non-detectable (< 4 microg/L) levels in acetate-fed columns inoculated with KJ or mixed cultures. The complete conversion of perchlorate to chloride was demonstrated by a stoichiometric ratio of perchlorate to chloride of 1.0 +/- 0.14. Perchlorate removal to non-detectable levels required a minimum empty bed contact time (EBCT) of only 2.1 min for the column inoculated with KJ, vs. 31 min for the mixed culture column. Acetate was used at a molar ratio of C2H3O2-/ClO4- of 2.9 (n = 6) for the mixed culture, while more than twice as much acetate was consumed on average (6.6 +/- 2.0, n = 156) by the pure culture. These results demonstrate that detention times of packed-bed bioreactors can be substantially reduced using isolate KJ, but that larger concentrations of acetate will be necessary to reduce perchlorate to low levels necessary for drinking water. PMID- 11487103 TI - Reductive dehalogenation of chlorinated ethenes with elemental iron: the role of microorganisms. AB - Trichloroethene (TCE) transformation and the product distribution in an aqueous medium containing zero-valent iron (Fe(0)) was investigated in the presence of an anaerobic mixed culture to assess the potential role of microorganisms in permeable iron barriers. The presence of the culture increased the rate of TCE disappearance and changed the product distribution. Rapid formation and degradation of cis-dichloroethene (cis-DCE) was observed in reactors containing cells plus Fe(0) or H2 as a bulk reducing agent. High levels of vinyl chloride (VC) were formed and very similar profiles were obtained in the Fe(0) plus cell and H2 plus cell reactors, but not in Fe(0)-only reactors. The similar trends observed in Fe(0)-cell and H2-cell reactors suggest that most cis-DCE and VC in the Fe(0)-cell reactors were produced and transformed biologically rather than abiotically. Accumulation of methane in the Fe(0)-cell system indicates that hydrogen gas generated during anaerobic iron corrosion could support a methanogenic culture. Digital confocal images showed that the microorganisms were able to colonize the iron surface. The results suggest that potential development of dechlorinating populations in Fe(0) barriers may alter the TCE reduction pathway and produce VC, which would have significant impact on the performance of Fe(0) barriers. PMID- 11487104 TI - The occurrence of E. coli O157:H7 in South African water sources intended for direct and indirect human consumption. AB - The occurrence of Escherichia coli O157:H7 in selected water samples in South Africa was investigated. The chromogenic Rainbow agar O157 medium designed for the rapid identification of E. coli O157:H7 was used for the detection of these organisms in various river-water samples in the Vaal Barrage Reservoir drainage basin of South Africa. A total of 204 samples were obtained from 15 sites where water was used for direct and indirect human consumption. Samples were filtered through Gelman filter-units and incubated on Rainbow agar O157 which produced different colours according to the bacterial chromogenic properties. Six hundred and sixty-three suspected E. coli O157:H7 colonies, with colours ranging between dark blue, grey and black, were subcultured onto sorbitol-MacConkey agar and screened for different virulence factors specific for E. coli O157:H7 and agglutination with anti-E. coli O157 antiserum. The results indicated that none of the suspected colonies contained all of the virulence factors necessary to classify them as E. coli O157:H7. None of these organisms agglutinated with antisera against E. coli O157. The probability of being infected with E. coli O157:H7 from direct or indirect consumption of these river water sources is therefore low. Some samples did, however, contain enterohaemorrhagic E. coli virulence properties, such as Stx1, Stx2 and enterohaemolysin, which might impose a health risk if ingested. PMID- 11487105 TI - Simultaneous removal of phenol and ammonia by an activated sludge process with cross-flow filtration. AB - Attempts were made for removing ammonia from synthetic wastewater under the presence of phenol, which is inhibitory to nitrification, by using a single-stage activated sludge process with cross-flow filtration. Activated sludge biomass which had been acclimated with phenol for over 15 years was used for the inoculum, and synthetic wastewater was continuously supplied to the process retaining biomass at 8000 mg VSS l(-1). Phenol was completely removed, and ammonia was simultaneously nitrified to nitrate; nitrification rate reached 200 mg N l(-1) d(-1) when phenol was removed at a rate up to 300 mg l(-1) d(-1). It was observed that 0-13% of the ammonia was removed via denitrification. Intermittent aeration enhanced the denitrification rate to 160 mg N l(-1) d(-1) by utilizing phenol. and approximately 24% of the denitrified nitrogen was recovered as nitrous oxide. Methanol, which is the most commonly used electron donor in conventional nitrogen removal processes, did not enhance the denitrification rate of the phenol-acclimated activated sludge used in this study, however phenol did. The results suggest that this process potentially works as a space- and energy-saving nitrogen removal process by utilizing substances inhibitory to nitrifiers as electron donors for denitrification. PMID- 11487106 TI - Biofilter treatment of aquaculture water for reuse applications. AB - Biotreatment of aquaculture water for recirculation purposes is a sensible mean to support the further growth of aquaculture industry without excessive water demands that are environmentally unsustainable. This study evaluates the efficacy of biofilter treatment of an eel (Anguilla japonica) culture pond water using different filter media and flow scheme arrangements. The experimental results demonstrate that biofilter systems packed with suitable filter media are capable of improving the quality of effluents for recirculation applications. The characteristics of the filter media appear to be more critical than biofilter flow scheme arrangements in affecting the efficacy of the biofilter treatment. Filter media with surface and structural characteristics are conducive to the development of biofilms and the capture of organic suspended matter are desirable to ensure good and consistent biofilter performance. Under such circumstances the bacterial "consortia" in the biofilter are capable of utilizing the captured organic suspended matter as an alternative substrate to support their metabolic activities when the concentration of the primary substrate (i.e., BOD) is low. For the eel pond water, a biofilter packed with filter media having cross-link structures and a high bed porosity, followed by another biofilter packed with a type of filter media having rough surfaces, produced the best results under the conditions tested. Moreover, a preliminary cost-benefit analysis confirms its cost advantages. PMID- 11487107 TI - Inactivation of bacteriophages in water by means of non-ionizing (UV-253.7 nm) and ionizing (gamma) radiation: a comparative approach. AB - Thc inactivation behaviour of the bacteriophages PHI X 174 (ssDNA virus). MS2 (ssRNA virus) and B40-8 (dsDNA) toward non-ionizing (UV-253.7 nm) as well as to ionizing radiation (gamma radiation) was studied in order to evaluate their potential as viral indicators for water disinfection by irradiation. Previous findings of the high UV-253.7 nm resistance of MS2 were confirmed whereas an unexpected high sensitivity to gamma radiation compared to the two other phages was found. On the other hand, PHI X 174 revealed an enhanced UV sensitivity but a high resistance to ionizing radiation. B40-8 had an intermediate position between the other two bacteriophages relative to both types of radiation. As expected, the data of E. coli reconfirmed the unreliability of fecal indicator bacteria for the purpose of predicting responses of viruses to water treatment. In UV disinfection the influence of water matrix may be adequately controlled by considering the UV (253.7 nm) absorption of the water whereas so far no such parameter has existed for the influence of the water quality on ionizing irradiation with respect to the scavenger concentration. PMID- 11487108 TI - Pollutant concentrations of stormwater and captured sediment in flood control sumps draining an urban watershed. AB - Levee sump systems are used by many riverine communities for temporary storage of urban wet weather flows. The performance and transport of stormwater pollutants in sump systems, however, have not been systematically studied. Although flow of stormwater through a sump system is regulated solely by flood-control requirements, these sumps may function as sedimentation basins that provide purification of stormwater. The objective of this paper is to present a case study of stormwater and sediment analysis for conventional pollutants in flood control sumps in an urban watershed. Two sumps of highly variable physical and hydraulic characteristics were selected for analysis. The results suggest that: (1) first-flush samples may not give an accurate picture of outfall concentrations of stormwater to the sump; (2) time-variable concentrations of pollutants in a sump are related to the hydraulic characteristics of the basin; and, (3) post-event sediment analysis provides verification of pollutant capture in the sumps. This study is an initial step toward development of a more comprehensive management model for flood control sumps that incorporates their hydraulic performance for quantitative assessment of pollutant loads, pollutant discharges, and pollutant removals with respect to the sumps. PMID- 11487109 TI - Integrated biological and physiochemical treatment process for nitrate and fluoride removal. AB - The feasibility of an integrated biological and physiochemical water treatment process for nitrate and fluoride removal has been evaluated. It consisted of two sequencing batch reactors (SBRs) in series. Performance of the process in the treatment of 24 synthetic water samples having nitrate concentrations of 40, 80, 120, 160, 200, and 250 mg/l (as N) and fluoride concentrations of 6, 10, 15, and 20 mg/l at different combinations was studied. Denitrification followed by defluoridation proved to be the best sequence of treatment. In all cases nitrate could be reduced to an acceptable level of less than 10 mg/l (as N) at 3, 5, and 7 h hydraulic retention times (HRTs) depending on its initial concentration. Fluoride concentrations up to 15 mg/l associated with nitrate concentrations up to 80 mg/l (as N) could be reduced acceptable 1.5 mg/l by alum-PAC slurry using alum doses up 850 mg/l [as Al2(SO4)3 x 16H2O] along with 100 mg/l of powdered activated carbon (PAC). Additional alkalinity produced during denitrification was used up during defluoridation for maintenance of pH avoiding the need for lime addition. On the other hand, residual organics, turbidity, and sulfide present in the denitrified water are removed by alum and PAC at the defluoridation stage along with fluoride, eliminating the need for an additional post-treatment step. At higher nitrate concentrations (> or = 120 mg/l as N), the alkalinity produced at the denitrification stage was in the range of 715-1175 mg/l as CaCO3. This excessive alkalinity inhibited reduction of fluoride to the level of 1.5 mg/l at the defluoridation stage, using alum doses up to 900 mg/l along with 100 mg/l of PAC. In all cases a fluoride concentration of 20 mg/l in water could not be reduced to the acceptable level of 1.5 mg/l. PMID- 11487110 TI - Hydraulic effects on sludge accumulation on membrane surface in crossflow filtration. AB - Membrane filtration technology for application of wastewater treatment has been developing recently. In the application to wastewater treatment, it is major concern to remove cake layer on membrane surface effectively with crossflow shear stress. Hydraulic effect of sludge accumulation process on membrane surface in bubble and non-bubble driven crossflow filtration was studied. Maximum sludge accumulation. sludge accumulation rate, and lag phase were introduced to describe sludge accumulation process, and the effects of hydraulic conditions were clarified experimentally. Maximum sludge accumulation and sludge accumulation rate were dependent on aeration intensity, and were less depend on flow channel width and MLSS concentration. Their tendencies were explained by shear stress. Shear stress was thought to be the major hydraulic factor that influences them. Lag phase was dependent on aeration intensity, flow channel width, and MLSS concentration. A non-dimensional equation was proposed to explain dependencies of flow channel width based on consideration of hydraulic behavior of MLSS particles and shear stress. PMID- 11487111 TI - Dye removal from textile dye wastewater using recycled alum sludge. AB - The removal of dyes from textile dying wastewater by recycled alum sludge (RAS) generated by the coagulation process itself was studied and optimized. One hydrophobic and one hydrophilic dye were used as probes to examine the performance of this process. It was found that RAS is a good way of removing hydrophobic dye in wastewater, while simultaneously reducing the fresh alum dosage, of which one third of the fresh alum can be saved. The back-diffusion of residued dye from the recycling sludge is detected but is easily controlled as long as a small amount of fresh alum is added to the system. The use of RAS is not recommended for the removal of hydrophilic dyes, since the high solubility characteristics of such dyes can cause deterioration in the water quality during recycling. PMID- 11487112 TI - Estimating biomass yield coefficients for autotrophic ammonia and nitrite oxidation from batch respirograms. AB - Kinetic characterization of biological processes via batch respirometry requires an accurate estimate of the biomass yield coefficient because it provides the stoichiometric link between biomass synthesis, substrate consumption and oxygen uptake. Expressions for biomass yield coefficients describing autotrophic ammonia and nitrite oxidation were derived from a mechanistically based electron balanced equation. We demonstrate that applying the conventional expression used to calculate the heterotrophic biomass yield results in erroneous estimates for the autotrophic biomass yield. Yield coefficients for autotrophic NH4(+)-N to NO2(-) N oxidation and NH4(+)-N to NO3(-)-N oxidation were overestimated by 27 to 36%. Due to correlation between the maximum specific growth rate and the biomass yield, the error in yield values propagated in 30 to 40% overestimates of the maximum specific growth rate coefficient for NH4(+)-N oxidation determined from batch respirograms. Therefore, it is essential to employ the correct expression to estimate the autotrophic biomass yield coefficient from batch respirograms due its inadvertent impact on subsequent parameter estimation. PMID- 11487113 TI - Lake ecosystem health assessment: indicators and methods. AB - A set of ecological indicators including structural, functional, and system-level aspects were proposed for a lake ecosystem health assessment, according to the structural, functional, and system-level responses of lake ecosystems to chemical stresses including acidification, eutrophication and copper, oil and pesticide contamination. The structural indicators included phytoplankton cell size and biomass, zooplankton body size and biomass, species diversity, macro- and micro zooplankton biomass, the zooplankton phytoplankton ratio, and the macrozooplankton microzooplankton ratio. The functional indicators encompassed the algal C assimilation ratio, resource use efficiency, community production, gross production/respiration (i.e. P/R) ratio, gross production standing crop biomass (i.e. P/B) ratio, and standing crop biomass unit energy flow (i.e. B/E) ratio. The ecosystem-level indicators conisisted of ecological buffer capacities, energy, and structural energy. Based on these indicators, a direct measurement method (DMM) and an ecological modeling method (EMM) for lake ecosystem health assessment were developed. The DMM procedures were designed to: (1) identify key indicators; (2) measure directly or calculate indirectly the selected indicators; and, (3) assess ecosystem health on the basis of the indicator values. The EMM procedures were designed to: (1) determine the structure and complexity of the ecological model according to the lake's ecosystem structure; (2) establish an ecological model by designing a conceptual diagram, establishing model equations, and estimating model pararmeters; (3) compare the simulated values of important state variables and process rates with actual observations; (4) calculate ecosystem health indicators using the ecological model; and, (5) assess lake ecosystem health according to the values of the ecological indicators. The results of a case study demonstrated that both methods provided similar results which corresponded with the lake's actual trophic state. PMID- 11487114 TI - In situ measurement and statistical modelling of Escherichia coli decay in small rivers. AB - Numerous studies have been carried out on the environmental factors associated with the decay of faecal bacteria in open (fresh or marine) waters. The present study aimed at understanding the fate of bacteria in small streams (flow <20 m3 s(-1)) for which there is a lack of knowledge. An original in situ protocol was developed for measuring the die-off of Escherichia coli (E. coli) from wastewater treatment plants. Based upon 80 values of the decay first-order parameter (K or its inverse T90), collected from five rivers in Normandy (France), a median T90 of 10 h and a minimal T90 of 1.3 h were obtained. K was then modelled as a linear function of variables made up from flow, water temperature and suspended particulate matter (SPM). The set of significant co-variables did not include light indicators. E. coli decay is inversely related to the river flow and it becomes highly significant below 0.3 m3 s(-1). The positive effect of small flows on die-off is increased by water temperature over 15 degrees C, whereas it could be reduced by SPM. The major co-variable of the model (p < 10(-9)) is an empiric composite variable integrating the effect of flow and temperature that explains more than 40% of the variance of K. We interpreted this as an expression of predation by benthic micro-grazers which could be the main cause of E. coli die off in small streams in temperate countries. PMID- 11487115 TI - Interaction between Cryptosporidium oocysts and water treatment coagulants. AB - The electrokinetic properties of gamma-irradiated Cryptosporidium oocysts in the presence of coagulants (ferric chloride and alum) and coagulant aids (DADMAC based cationic polyelectrolytes) have been studied. The zeta potential of the oocysts was unaffected by the addition of ferric chloride at all pH values (3-10) studied. Addition of alum resulted in reversal of the oocysts charge, which suggests that the initial stage in the coagulation process leading to floc formation proceeds via the adsorption of hydrolysed aluminium species. The cationic polyelectrolyte Magnafloc LT35 was adsorbed onto iron flocs at doses of 0.1 mg/L even against an electrostatic barrier. The cationic polyelectrolyte only adsorbed and caused charge reversal at the oocyst surface at around 0.4 mg/L, suggesting a lower affinity for this surface. These results indicate that the oocysts, unlike inorganic colloidal materials such as metal oxides, appear to possess a lower surface density of active or charged sites. The lower density of sites, combined with the rapid precipitation of iron salts, may be responsible for the lack of specific adsorption of either hydroxylated ferric species or primary iron hydroxide particles on the oocysts. Further, this suggests that a process of sweep flocculation, where oocysts are engulfed in flocs during coagulation and floc formation, is the more likely mechanism involved. By comparison, it is likely that the specific interaction of hydrolysed aluminium species with the oocysts surface would result in a stronger link at the oocyst floc interface and that the flocculation process may initially proceed via charge neutralisation. PMID- 11487116 TI - Stability of enhanced biological phosphorus removal and composition of polyphosphate granules. AB - The influence of varying Ca- and Mg-concentration of the influent wastewater on the enhanced biological phosphorus removal was investigated in an anaerobic aerobic bench-scale plant. The artificial enhancement of the Mg-concentration in the influent from 15 to 24 mg l(-1) and 31 mg l(-1), respectively, caused a raise of the mean P-removal efficiency from 85 to 97%. The P-elimination was very stable in time. A chemical precipitation of magnesium ammonium phosphate could be excluded. The elemental composition of polyphosphate granules was investigated by electron microscopy and energy dispersive X-ray spectroscopy. The elements Ca, Mg and K were the principal metal components of polyphosphate granules. Concerning the metal composition, different types of granules could be distinguished. The quantitative ratios of Ca, Mg and K varied in dependence on the influent concentration of these metals. A relation between the Mg/Ca-ratio of the granules and the efficiency of enhanced biological phosphorus removal can be supposed. PMID- 11487117 TI - Selective degradation of ibuprofen and clofibric acid in two model river biofilm systems. AB - A field survey indicated that the Elbe and Saale Rivers were contaminated with both clofibric acid and ibuprofen. In Elbe River water we could detect the metabolite hydroxy-ibuprofen. Analyses of the city of Saskatoon sewage effluent discharged to the South Saskatchewan river detected clofibric acid but neither ibuprofen nor any metabolite. Laboratory studies indicated that the pharmaceutical ibuprofen was readily degraded in a river biofilm reactor. Two metabolites were detected and identified as hydroxy- and carboxy-ibuprofen. Both metabolites were observed to degrade in the biofilm reactors. However, in human metabolism the metabolite carboxy-ibuprofen appears and degrades second whereas the opposite occurs in biofilm systems. In biofilms the pharmacologically inactive stereoisomere of ibuprofen is degraded predominantly. In contrast, clofibric acid was not biologically degraded during the experimental period of 21 days. Similar results were obtained using biofilms developed using waters from either the South Saskatchewan or Elbe River. In a sterile reactor no losses of ibuprofen were observed. These results suggested that abiotic losses and adsorption played only a minimal role in the fate of the pharmaceuticals in the river biofilm reactors. PMID- 11487118 TI - Numerical and graphical description of the information matrix in calibration experiments for state-space models. AB - This paper describes a mathematical tool for local identifiability analysis that can easily be applied to high-order state-space nonlinear systems and implemented in simulators with a discrete-time approach. The methodology is based on the recursive numerical evaluation of a reduced information matrix during the simulation of a calibration experiment and in the setting-up of a group of information parameters based on geometric interpretations of this matrix. As an example of application, the proposed methodology has been used in the study of an OUR batch test from the point of view of ASM No. 1 calibration. PMID- 11487119 TI - Development of the phytoplankton community in a pit-lake in relation to water quality changes. AB - An open pit-lake was formed at a Northern Saskatchewan mine site after flooding with lake water in early 1992. Since then, water and phytoplankton samples have been collected regularly from the artificial lake over seven years. The resulting data set provides a unique opportunity to examine the physical and chemical changes in water quality and phytoplankton community over time. Seventeen major variables were examined in a principal component analysis. Axes 1, 2 and 4 are significantly correlated to other three variables, the number of days (since the first sampling after flooding of the pit), water temperature and depth, respectively. Total suspended solid (TSS), dissolved oxygen, total phosphorus (Total-P), arsenic, and iron decreased over time while Mg, Ca, K, Na, total organic carbon and HCO3 increased. Canonical correspondence analysis was used to infer the relationship between water quality variables and phytoplankton community structure, which changed substantially over the survey period. TSS, Total-P and arsenic were considered to be the key factors driving the change in phytoplankton community composition. PMID- 11487120 TI - Hydrodynamic drag on non-spherical floc and free-settling test. AB - This work numerically investigates the hydrodynamic drag force exerted on a porous spheroid floc moving steadily through a quiescent Newtonian fluid over a wide range of the Reynolds number. The flow patterns for a highly porous spheroid moving at an elevated Reynolds number are basically the same as those at a low Reynolds number, which extends the applicable range of a creeping-flow based correlation to the higher Reynolds number regime. The shape effect becomes more prominent as the spheroid becomes more porous. Using the equivalent diameter, defined as the geometric mean diameter of the principal axes, leads to a universal correlation relating to the drag force, aspect ratio, and interior permeability. In addition, free-settling experiments are performed to estimate how the non-spherical shape affects the three sludge samples. The possible errors in data reduction for the free-settling test are attributed to the a/b ratio and the internal permeability. The errors range from 16-34% for a/b = 0.6-2.0. PMID- 11487122 TI - Improved AOX degradation in UV oxidative waste water treatment by dialysis with nanofiltration membrane. AB - In this article, the wastewater treatment by UV oxidation with and without preceding desalination is compared. The influence of different chloride concentrations on the TOC degradation and AOX concentration is analyzed. Nanofiltration membrane dialysis is used to separate the chloride ions from wastewater. It is demonstrated that a reduction of the chloride concentration leads to a faster TOC degradation compared to the treatment of non-desalinated wastewater. Furthermore, the additional formation of AOX during the process could be avoided in effect leading to a significant degradation of native AOX. PMID- 11487121 TI - Oxidising and disinfecting by hydrogen peroxide produced in a two-electrode cell. AB - Hydrogen peroxide was produced by direct current electrolysis using two electrodes only, a carbon felt cathode and a dimensional stabilised anode (titanium coated with RuO2), without adding any chemical. The required oxygen was supplied by water oxidation and by transfer from the atmosphere. The intensity should be maintained under a maximum value to avoid peroxide reduction. High peroxide production rate and concentration were then reached. Electroperoxidation partially removed dissolved organic carbon (DOC) contained in solutions of phenol, salicylic acid, benzoic acid and humic acids. The DOC removal in effluent of municipal sewage plant corresponded to a breakage of the double bonds. Real effluents were significantly disinfected owing to the direct effect of electric current and the indirect effect of peroxide. Moreover, a remnant effect was ensured. PMID- 11487123 TI - Magnetic water treatment for scale prevention. AB - A home-made magnetic device was built with permanent magnets for treating scaling waters. Its efficiency was evaluated by measuring the remaining ionic calcium at the output of the device by means of an ion selective electrode. The scaling power of the treated water was estimated through an electrochemical scaling test. Chroamperometric curves and chronoelectrogravimetric curves were plotted to obtain the scaling time and the nucleation time of the scale deposition. The variation of the efficiency of the magnetic treatment was studied when the length of treatment, the flow velocity of the scaling water in the device, the material of the pipe where the scaling water flowed were changed. An empirical relationship, which gives the value of the efficiency in function of the length of treatment and the flow velocity, was proposed. Possible mechanisms of action of the magnetic treatment were discussed. PMID- 11487124 TI - Selective demineralization of water by nanofiltration application to the defluorination of brackish water. AB - Nanofiltration is generally used to separate monovalent ions from divalent ions, but it is also possible to separate ions of the same valency by careful application of the transfer mechanisms involved. Analysis of the retention of halide salts reveals that small ions like fluoride are the best retained, and that this is even more marked under reduced pressure when selectivity is greatest. The selectivity desalination of fluorinated brackish water is hence feasible and drinking water can be produced directly at much lower cost than using reverse osmosis by optimizing the pressure for the type of water treated. PMID- 11487125 TI - The BAC-process for treatment of waste water containing non-ionogenic synthetic surfactants. AB - In this paper experimental results on the biological-activated carbon (BAC) process for biological degradation and adsorption of non-ionogenic surfactants (NISS) in the waste water treatment are discussed. It is shown that the mechanism of the BAC-process is not the simple addition but the synergetic combination of biodegradation and carbon adsorption. The major aspects of such synergism are the biological regeneration (bioregeneration) of the adsorbent and the reduction of the toxic effect of waste water contaminants on microorganisms. It is shown that the basis of the bioregeneration process is the desorption of substances previously adsorbed on the activated carbon. The desorption from micropores takes place because of the reverse concentration gradient, due to the microbial degradation of waste water contaminants in the liquid phase. The desorption from mesopores is also supported by the activity of microorganisms exoenzymes. Thus, the process of bioregeneration is featured by two non-contradictory hypotheses. PMID- 11487126 TI - Atrazine degradation under denitrifying conditions in continuous culture of Pseudomonas ADP. AB - The simultaneous removal of atrazine and nitrate in continuous culture under denitrifying conditions using Pseudomonas sp. strain ADP was investigated. Under all operational conditions the nitrate removal efficiency was always higher than 90%, while atrazine degradation deteriorated with time due to contamination by foreign denitrifying bacteria, lacking the ability to degrade atrazine. Recovery of atrazine degradation ability was achieved by applying aerobic conditions with atrazine as the sole nitrogen source. PMID- 11487127 TI - Micellar effect on the photolysis of hydrogen peroxide. AB - Photolysis experiments were performed to quantify the effect of three anionic surfactants on the photolysis of hydrogen peroxide (H2O2) at the ambient laboratory temperature of 22+/-1 degrees C. H2O2 photolysis in water, methanol, and surfactant monomeric solution was also conducted to compare the photochemical reactivity of H2O2 in different media. Photolysis rates were highest for water, followed by micellar solutions, and lowest for methanol. The results show that the photochemical reactivity of H2O2 is less favorable in organic solvent than in water and surfactant micelles affect H2O2 photolysis. Retarded photolysis of H2O2 in micellar solutions implies that a fraction of H2O2 dissolved in water partitions into micellar pseudophase of surfactant. H2O2 partitioned into micelles has less photochemical reactivity and thus photolysis rate was retarded in the presence of micelles. Photolysis inhibitory level by micelles was shown to be dependent on the kinds of surfactants used in this study. In addition, the inhibitory effect by surfactant monomers was negligible due to the absence of micelles. PMID- 11487128 TI - Comment on "Estimating the infection risk in recreational waters from the faecal indicator concentration and from the ratio between pathogens and indicators". PMID- 11487129 TI - Measurement of intraosseous pressures generated by the Wand, high-pressure periodontal ligament syringe, and the Stabident system. AB - Intraosseous pressure generated by the use of three anesthetic systems-the Wand; a hand-operated high-pressure periodontal ligament (PDL) syringe; and the Stabident system-were studied in fresh mandibles of 14 large swine. The mandibles were drilled and tapped in one area of both the right and left posterior molar regions. Pressure gauges were attached via threaded fittings. Pressures during injection were recorded for the Wand first, then the PDL syringe, and finally Stabident. Results showed averages of 8.3 mm Hg generated by the Wand, 16.3 mm Hg with the high-pressure PDL syringe, and 43.7 mm Hg from the Stabident system. Results were corroborated with data from three human cadaver mandibles. PMID- 11487130 TI - An immunohistological study on cyclooxygenase-2 in human dental pulp. AB - Characteristics of cyclooxygenase-2 (COX-2) expressing cells in human dental pulp were immunohistologically studied. Extirpated pulpal tissues from extracted teeth were examined to elucidate the localization and distribution of COX-2. Pulpal tissues were examined by the labeled streptavidin biotin method using specific mouse monoclonal antibodies for COX-2. Cell types of the COX-2 expressing cells were also investigated by the double stain technique using both monoclonal antibodies for CD68/macrophage and anti-COX-2. COX-2 expressing cells could be found in all of the inflamed pulps, and these cells were mostly distributed close to the area of accumulation of inflammatory cells. COX-2 was mainly expressed in fibroblasts rather than macrophages. In contrast, COX-2 expressing cells were scarcely found in the normal pulps. These findings indicate that pulpal fibroblasts, as well as macrophages, may participate in the production of prostaglandin through COX-2 expression in pulpal inflammation, and might be involved in the pathogenesis of irreversible pulpitis. PMID- 11487131 TI - Phase, compositional, and morphological changes of human dentin after Nd:YAG laser treatment. AB - Although techniques for repairing root fracture have been proposed, the prognosis is generally poor. If the fusion of a root fracture by laser is possible, it will offer an alternative to extraction. Our group has attempted to use lasers to fuse a low melting-point bioactive glass to fractured dentin. This report is focused on the phase, compositional, and morphological changes observed by means of X-ray diffractometer, Fourier transforming infrared spectroscopy, and scanning electron microscopy-energy dispersive X-ray spectroscopy in human dentin after exposure to Nd:YAG laser. The irradiation energies were from 150 mJ/ pulse-10 pps-4 s to 150 mJ/pulse-30 pps-4 s. After exposure to Nd:YAG laser, dentin showed four peaks on the X-ray diffractometer that corresponding to a-tricalcium phosphate (TCP) and beta-TCP at 20 = 30.78 degrees/34.21 degrees and 32.47 degrees/33.05 degrees, respectively. The peaks of a-TCP and beta-TCP gradually increased in intensity with the elevation of irradiation energy. In Fourier transforming infrared analysis, two absorption bands at 2200 cm(-1) and 2015 cm(-1) could be traced on dentin treated by Nd:YAG laser with the irradiation energies beyond 150 mJ/pulse 10 pps-4 s. The energy dispersive X-ray results showed that the calcium/phosphorus ratios of the irradiated area proportionally increased with the elevation of irradiation energy. The laser energies of 150 mJ/ pulse-30 pps-4 s and 150 mJ/pulse-20 pps-4 s could result in the a-TCP formation and collagen breakdown. However, the formation of glass-like melted substances without a-TCP at the irradiated site was induced by the energy output of 150 mJ/ pulse-10 pps-4 s. Scanning electron micrographs also revealed that the laser energy of 150 mJ/ pulse-10 pps-4 s was sufficient to prompt melting and recrystallization of dentin crystals without cracking. Therefore, we suggest that the irradiation energy of Nd:YAG laser used to fuse a low melting-point bioactive glass to dentin is 150 mJ/ pulse-10 pps-4 s. PMID- 11487132 TI - Aspartate aminotransferase activity in human healthy and inflamed dental pulps. AB - Aspartate aminotransferase (AST) seems to be an important mediator of inflammatory processes. Its role in the progression and detection of inflammatory periodontal disease has been increasingly recognized in recent years. In the present study AST activity was analyzed in normal healthy human dental pulps, in reversible pulpitis, and in irreversible pulpitis. Enzymatic AST activity showed that the control values for the healthy pulps were 4.8 +/- 0.7 units/mg of pulp tissue. In reversible pulpitis specimens the AST activity increased to 7.98 +/- 2.1 units/mg of pulp tissue. In irreversible pulpitis specimens the values decreased to 2.28 +/- 1.7 units/mg of pulp tissue. Differences between the groups (control versus reversible pulpitis and reversible pulpitis versus irreversible pulpitis) were statistically significant (p = 0.0015). These results could point to a role of AST in the early events that lead to development of pulpal inflammation. PMID- 11487133 TI - Formaldehyde release from ground root canal sealer in vitro. AB - The formaldehyde release from three different ground root canal sealer materials was examined. Ten specimens each of AH26, Amubarut, and N2 were stored under dry conditions for 6 months. An amount of approximately 100 to 200 mg ground material was obtained from each sample by using a round bur and stored for 10 min in distilled water. The formaldehyde concentration of the immersion water was determined by high-performance liquid chromatography. The mean formaldehyde release per mg material was 6.6 (+/-2.5) microg for AH26 and 8.3 (+/-1.0) microg for Amubarut. A lower formaldehyde release was detectable by our method from the N2 samples (0.3 +/- 0.1 microg/g; p < 0.0001). In conclusion formaldehyde release from ground root canal material is low, although a risk of an allergic reaction in susceptible patients cannot be excluded. PMID- 11487134 TI - Bacterial penetration and proliferation in root canal dentinal tubules after applying dentin adhesives in vitro. AB - Endodontic treatment is aimed at eliminating infection and preventing bacterial regrowth in the root canal and dentinal tubules. In the present study the ability of two dentin adhesives to prevent bacterial penetration and subsequent proliferation in dentinal tubules was evaluated. Cylindrical root specimens prepared from freshly extracted bovine teeth were used in an in vitro model of dentinal tubule infection. After removal of the smear layer the intracanal dentinal tubules of the specimens were acid-etched and treated with either Gluma or EBS. Untreated specimens served as controls. Specimens were infected with Enterococcus faecalis and incubated in Brain Heart Infusion for 21 days. Powder dentin samples obtained from within the canal lumina, using ISO 025 to 033 burs, were examined for the presence of vital bacteria by inoculating on agar plates and counting colony-forming units. A significant difference was found between the experimental groups and the untreated group. After application with Gluma specimens showed the least viable bacteria in dentinal tubules. Data suggested that dentin adhesives reduced bacterial invasion into dentin and therefore have a potential role in endodontic treatment. PMID- 11487135 TI - Effect of sodium hypochlorite and five intracanal medications on Candida albicans in root canals. AB - The aim of this study was to evaluate the effect of 1% sodium hypochlorite and five intracanals medications on Candida albicans harvested inside root canals. The contaminated canals were irrigated with sterile saline solution and then treated as follows: (i) filled with Calen paste (calcium hydroxide/ glycol polyethylene paste); (ii) filled with camphorated paramonochloro phenol (CPMC); (iii) filled with 2% iodine-iodate solution; (iv) filled with tricresol formalin; (v) filled with Calen and CPMC pastes; (vi) irrigation with 1% sodium hypochlorite and filled with no intracanal medication; and (vii) no intracanal medication was used. Canal access and the apical foramen were then sealed with Cavit and the roots were stored in a humid chamber at 37 +/- 1 degree C for 14 days. The canals were reinstrumented and irrigated with sterile saline solution. Sterile paper points were used to transfer the root canal contents to test tubes containing sterile saline solution. Part of the suspension was harvested in Sabouraud dextrose agar with chloramphenicol and incubated at 37 +/- 1 degree C for 48 h. CPMC was effective in 100% of the samples followed in decreasing order of effectiveness by calcium hydroxide with CPMC (70% effective), 1% sodium hypochlorite (70% effective) (p < 0.05), tricresol formalin (60% effective), 2% iodine-iodate solution (50% effective), calcium hydroxide paste (30% effective), and saline + no intracanal medication. PMID- 11487136 TI - Histologic assessment of healing after the use of a bioresorbable membrane in the management of buccal bone loss concomitant with periradicular surgery. AB - Histological and morphometric assessment of periradicular wound healing was made after the use of a bioresorbable membrane over a buccal dehiscence. The third and fourth premolar teeth of nine dogs were resected and buccal defects created. Teeth were assigned randomly to the membrane or control group. One tooth in each quadrant received a membrane, covering both roots. The other tooth received no further treatment and served as a control. The animals were killed and specimens were assessed at two time periods: 9 wk and 27 wk. The 27-wk membrane group exhibited significantly more (p = 0.004) connective tissue height than the control group or either of the 9-wk groups. The amount of regenerated alveolar bone was significantly greater for the 27-wk membrane group than for the control (p = 0.001) and 9-wk groups. Mean junctional epithelium measurements were significantly greater (p = 0.012) for the control. The use of a bioresorbable membrane enhances bone regeneration when a buccal defect exists at the time of periradicular surgery. PMID- 11487137 TI - Effects of eugenol on resin bond strengths to root canal dentin. AB - The purpose of this study was to compare the regional bond strengths of C&B Metabond resin to root canal dentin, with or without treatment using a eugenol containing endodontic sealer liquid. Eighteen extracted human canines were decoronated at the cementoenamel junction with a slow speed saw. The apical third of the root was removed leaving the cervical and middle dentin. The canal space was then enlarged with files, Gates-Glidden burs, and parapost drills. The teeth were ground on either the mesial or distal sides, permitting direct access to the entire canal. The cervical or middle third dentin was treated with Kerr Root Canal Sealer liquid, alternating between the middle and cervical thirds. Each tooth served as its own control. The adhesive resin was then luted directly to the prepared canal. Specimens, 1 x 1 x 8 mm, were prepared and mounted to a Vitrodyne testing machine enabling microtensile bond strengths to be measured. Data were analyzed using a two-way ANOVA and the least squares means test. The mean microtensile bond strengths for the cervical and middle third dentin treated with eugenol were 13.6 +/- 6.1 MPa (n = 33) and 14.8 +/- 3.9 MPa (n = 29), respectively. Without the eugenol, the mean bond strengths were 18.1 +/- 6.0 MPa (n = 31) and 17.3 +/- 4.6 MPa (n = 31) for the cervical and middle sections. The specimens treated with the eugenol liquid had significantly lower bond strengths than those without eugenol (p < 0.05) only in the cervical third. The region of the tooth tested had no effect on bond strength. That is, bond strength of the cervical third was not significantly different from bond strength on the middle third in either of the two groups (with or without eugenol) tested. PMID- 11487138 TI - Effect of apical trephination on postoperative pain and swelling in symptomatic necrotic teeth. AB - The purpose of this prospective, randomized, blinded study was to determine the effect of apical trephination on postoperative pain and swelling in symptomatic necrotic teeth. Fifty emergency patients participated, and each had a clinical diagnosis of a symptomatic necrotic tooth with associated periapical radiolucency. After endodontic treatment, patients randomly received either an apical trephination or mock trephination procedure. The trephination procedure used a Stabident perforator to provide an initial opening in the cortical bone that was enlarged with files (#25 through #120) and an endodontic spoon. Postoperatively, each patient received: ibuprofen; acetaminophen with codeine (30 mg); and a 7-day diary to record pain, percussion pain, swelling, and number and type of pain medication taken. Results demonstrated the use of an apical trephination procedure did not significantly (p > 0.05) reduce pain, percussion pain, swelling, or number of ibuprofen tablets taken in symptomatic necrotic teeth with periapical radiolucencies. The trephination procedure did significantly (p < 0.05) reduce the use of acetaminophen with codeine overall for the 7 days. In conclusion, because there was not a significant reduction in pain, percussion pain, or swelling we cannot recommend the routine use of an apical trephination procedure, as used in this study, in symptomatic necrotic teeth with radiolucencies. PMID- 11487139 TI - Setting times for endodontic sealers under clinical usage and in vitro conditions. AB - The object of this study was to determine the set of sealers under simulated clinical versus bench-top conditions. One hundred twenty extracted teeth were divided into four groups (Roth's, Tubuliseal, Sealapex, AH26). Canal preparation and lateral condensation were performed with the four sealers (30 teeth each). Teeth were placed into 100% humidity at 37 degrees C. The same mix of sealer on a glass slab was stored within the same environment. After 1 to 8 wk the teeth were fractured longitudinally to expose the gutta-percha-sealer interface. Evaluation, both clinical and in vitro, was qualitative under a microscope with a needle tip. The degree of set was categorized as: (a) unset (smearable), (b) partially set (indented easily, but not smearable), or (3) set (not easily indentable). There were inconsistencies in setting times within and between groups. In canals most of three (AH26, Sealapex, and Tubuliseal) were partially set after 1 wk; set was complete after 4 wk. Roth's was very slow; most were unset after 8 wk. Sealers on the glass slab set much more rapidly. In conclusion, under clinical conditions, sealers set slowly (particularly Roth's) and were more delayed than when tested in vitro. PMID- 11487140 TI - Zebra XX, part 2. PMID- 11487141 TI - Ask a friend. Case 11. PMID- 11487142 TI - The effect of calcitonin on osseous healing in guinea pig mandible. AB - Calcitonin (CT) is reported to be an effective medication for the treatment of inflammatory root resorption and to be capable of stimulating osteoblast proliferation in cell culture studies. In this study the effect of CT on the healing of osseous defects was evaluated in the mandibles of guinea pigs. After raising tissue flaps two experimental cavities were created on both sides of the corpus mandible of 33 guinea pigs. CT was applied into cavities either in hydroxypropyl methycellulose (HPMC) gel or gelatin as carrier. HPMC and gelatin alone and an empty cavity were also examined as control groups. Histopathological examinations under light microscopy were performed on weeks 1, 3, and 6. At week 1 in CT+gelatin and CT+HPMC groups, prominent osteoblastic activity was observed when compared with control groups. At week 3 the presence of woven bone in the experimental cavity areas reflected the increased osteoblastic activity in all groups. At the end of week 6 woven bone was gradually replaced by osteogenic tissue undergoing remodelization with Haversian systems in all groups. It is suggested that the osseous healing of the experimental cavity was enhanced by CT application in early stages (i.e. at week 1). However there was no significant difference of osteogenic activity between the control and CT-treated groups at the end of weeks 3 and 6. PMID- 11487143 TI - Detection of putative oral pathogens in acute periradicular abscesses by 16S rDNA directed polymerase chain reaction. AB - A 16S rDNA-directed polymerase chain reaction method was used to assess the occurrence of four black-pigmented anaerobic rods, Treponema denticola, and Actinobacillus actinomycetemcomitans in acute periradicular abscesses. Pus was collected by aspiration from 10 cases diagnosed as acute abscesses of endodontic origin. DNA was extracted from the samples and analyzed using a polymerase chain reaction-based identification assay. The method allowed detecting black-pigmented anaerobes in 80% of the examined abscesses. Porphyromonas endodontalis was found in 70%, T. denticola in 50%, Porphyromonas gingivalis in 40%, and Prevotella intermedia in 10% of the cases. P. gingivalis was always found associated with P. endodontalis. Prevotella nigrescens and A. actinomycetemcomitans were not found in any pus sample. The high prevalence of P. endodontalis, T. denticola, and P. gingivalis suggests that they can play an important role in the etiology of acute periradicular abscesses. PMID- 11487144 TI - Adhesion of a glass-ionomer root canal sealer to the root canal wall. AB - Glass-ionomer root canal sealer is commonly used because of its chemical bonding and favorable physical characteristics when bonding to dentin. This study was designed to determine the tensile bond strength of a glass-ionomer sealer (Ketac Endo, Espe, Seefeld, Germany) on root canal walls after pretreatment with different conditioners. Flat inner surfaces of root canal specimens were prepared. The specimens were divided into five groups of 10 teeth, and the groups were conditioned with one of the following smear layer removal solutions: 15% EDTA/NaOCl, 10% polyacrylic acid, 35% phosphoric acid, 6% citric acid, and 5.25% NaOCl as a control. Then the exposed root canal areas were coated with Ketac Endo. Tensile bonding was measured using a universal testing machine until ultimate failure was obtained. The groups that were treated with phosphoric acid and citric acid showed significantly higher bond strengths than the groups that were treated with 15% EDTA and polyacrylic acid (p < 0.05). Bonding to dentin without smear layer removal (NaOCl group) was too low to be measured in the testing apparatus. Scanning electron microscopy confirmed that phosphoric and citric acids were more effective in removing smear layer than EDTA or polyacrylic acid. The result supported the view that pretreatment with phosphoric acid or citric acid should be used in association with glass-ionomer root canal sealer to achieve the most effective removal of the smear layer and to provide better adhesion. PMID- 11487145 TI - Antibacterial properties of electron beam-sterilized Gutta-Percha cones. AB - The purpose of the study was to determine the effect of electron beam sterilization on gutta-percha cones (GPCs) at different times after sterilization. An agar diffusion test was used with -one aerobic bacterium (Bacillus subtilis) and five oral anaerobic bacteria (Fusobacterium nucleatum, Peptostreptococcus micros, Porphyromonas gingivalis, Propionibacterium acnes, and Veillonella parvula). With each microorganism 30 agar plates were prepared, evenly distributed among three groups (group 1: unsterilized GPCs; groups 2 and 3: GPCs sterilized by electron beam irradiation 5 months and 5 yr before, respectively). One GPC of the selected group was placed in each plate. After incubation the area of inhibition was read on the agar plates. Inhibition of growth was significantly different for the tested microorganisms. However no significant difference was observed between the sterilized and unsterilized GPCs. Both the tested sterilized and unsterilized GPCs impair the growth of endodontic pathogens, with no influence of the time elapsed since sterilization. PMID- 11487146 TI - Stimulation of plasminogen activator activity and matrix metalloproteinases of human dental pulp-derived cells by tumor necrosis factor-alpha. AB - The effects of tumor necrosis factor (TNF)-alpha on the plasminogen activator (PA)/plasmin system in human dental pulp (HDP) cells were examined. TNF-alpha treatment induced a significantly high level of PA activity in the conditioned medium of HDP cells in a time- and dose-dependent manner, compared with untreated control cells. Western-blot analysis revealed that tissue type (t)PA protein in conditioned medium was increased by TNF-alpha when compared with control medium. Furthermore the tPA mRNA level had increased in HDP cells treated with TNF-alpha, as determined by reverse transcription-polymerase chain reaction, but urokinase PA and PA inhibitor-1 mRNA levels did not increase. We examined the effects of TNF-alpha against activities of matrix metalloproteinases (MMPs) using zymography. TNF-alpha stimulated MMP-2 activity in conditioned medium and stimulated MMP-9 activity with addition of plasminogen into conditioned medium. The present results suggested that TNF-alpha stimulates PA activity via an enhancement of tPA gene expression in HDP cells and MMP-2 activity, and further that tPA-activated TNF-alpha stimulated MMP-9. PMID- 11487147 TI - Alkaline phosphatase activity in normal and inflamed dental pulps. AB - Alkaline phosphatase (ALP) seems to be important in the formation of mineralized tissues. High levels of ALP have been demonstrated in dental pulp cells. In the present study ALP activity was analyzed in normal healthy human dental pulps, in reversible pulpitis, and in irreversible pulpitis. Enzymatic ALP control values for the normal healthy pulps were 110.96+/-20.93. In the reversible pulpitis specimens the ALP activity increased almost eight times to 853.6+/-148.27. In the irreversible pulpitis specimens the values decreased sharply to 137.15+/-21.28 and were roughly equivalent to those seen in normal healthy pulps. The differences between the groups (control vs. reversible pulpitis and reversible pulpitis vs. irreversible pulpitis) were statistically significant. These results could point to a role of ALP in the initial pulp response after injury. PMID- 11487148 TI - Evaluation of the effect of EDTAC, CDTA, and EGTA on radicular dentin microhardness. AB - The effect of EDTAC (ethylenediamine tetra-acetic acid plus Cetavion, an ammonium surfactant), CDTA (cyclohexane-1,2-diaminetetra-acetic acid), and EGTA (ethylene glycol-bis-(beta-amino-ethyl ether) N,N,N',N'-tetra-acetic acid) on the microhardness of radicular dentin of the cervical third of human teeth was studied. Five newly extracted maxillary incisors were sectioned transversely at the cementoenamel junction, and the crowns were discarded. The roots were embedded in blocks of high-speed polymerized acrylic resin and cut transversely into 1-mm sections. The second slice of the cervical third of the root of each tooth was sectioned and divided into four parts. Each part was placed on an acrylic disc that was used as a base for microhardness measurement. Fifty microliters of 15% EDTAC, 1% CDTA, or 1% EGTA were applied to the dentin surface. Deionized and distilled water was used as control. Dentin microhardness was then measured with a Vicker's microhardness apparatus with a load of 50 g for 15 s. Statistical analysis showed that the three chelating solutions significantly reduced dentin microhardness when compared with water; however, there was no statistically significant difference among the three solutions. PMID- 11487149 TI - Induction of dental pulp fibroblast matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 gene expression by interleukin-1alpha and tumor necrosis factor-alpha through a prostaglandin-dependent pathway. AB - Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) are involved in the degradation of extracellular matrix in many inflammatory diseases. Little is known regarding the expression of these mediators in dental pulp fibroblasts. The effects of proinflammatory cytokines (interleukin (IL)-1alpha and tumor necrosis factor-alpha (TNF-alpha)) and prostaglandin E2 (PGE2) on pulp fibroblast MMP-1 and TIMP-1 gene expression were investigated. Northern hybridization showed that IL-1alpha and TNF-alpha induced significant MMP-1 gene expression, with only little effect on TIMP-1 gene. Exogenous PGE2, however, upregulated TIMP-1 mRNA synthesis but not MMP-1. Concomitant addition of IL-1alpha and PGE2 or TNF-alpha and PGE2 suppressed MMP-1 mRNA production, compared with the groups treated with IL-1alpha or TNF-alpha alone. In contrast, PGE2 enhanced the upregulatory effects of TIMP-1 mRNA by IL 1alpha or TNF-alpha. Furthermore, cytokine stimulation of MMP-1 and TIMP-1 gene expressions can be enhanced or blocked by indomethacin, respectively, and reversed by exogenous PGE2. These results suggested that cytokine-stimulated MMP 1 and TIMP-1 gene expression in dental pulp fibroblasts was mediated, at least in part, by a prostaglandin-dependent pathway. The differential regulation of IL 1alpha or TNF-alpha-induced MMP-1 and TIMP-1 mRNA synthesis, as well as the direct upregulation of TIMP-1 gene expression by PGE2, also implied that prostaglandin may serve as a protective mechanism from excessive tissue breakdown during pulpitis. PMID- 11487150 TI - The anti-inflammatory effects of human recombinant copper-zinc superoxide dismutase on pulp inflammation. AB - Inflammation in the dental pulp is accompanied by release of a wide variety of highly oxidative molecules known as reactive oxygen species (ROS). ROS concentrations are controlled in vivo by an antioxidant enzyme scavenger system that may be overwhelmed by the increases in ROS production seen during inflammation. Supplementation of the antioxidant defense system, therefore, may limit the severity of the inflammatory response to injury due to this component. To test this hypothesis, this study examined the effects of superoxide radical scavenging on pulpal inflammation induced in rat molars by standardized cavity preparation. The extent of pulp inflammation was compared histomorphometrically between animals treated with exogenous administration of a human recombinant antioxidant enzyme, copper-zinc superoxide dismutase, conjugated to polyethylene glycol (hr-CuZn-SOD), versus saline-vehicle controls. There was a statistically significant reduction in area of inflammation involvement in those animals treated with hrCuZn-SOD, compared with controls. Although hrCuZn-SOD administration did not completely eliminate inflammation in all animals treated, there was a statistically significant lessening of the severity of the inflammatory response, as well as a greater degree of reparative dentin observed in the hrCuZn-SOD-treated animals. PMID- 11487151 TI - Comparing apical preparations of root canals shaped by nickel-titanium rotary instruments and nickel-titanium hand instruments. AB - This study measured displacement of canal centers in extracted human teeth after preparation by Lightspeed and nickel-titanium K-files. The specimens were divided into two groups (n = 11), after radiographing the roots from the buccolingual and mesiodistal planes with pathfinding files in situ. More strongly and less strongly curved canals were, respectively, assigned to the Lightspeed and nickel titanium K-file groups. Using a reassembly technique, cross-sections at 1.25 mm, 3.25 mm, and 5.25 mm from the apices were made of the experimental roots embedded in clear plastic. Stereomicroscopic 35-mm slides were taken of the uninstrumented sectioned canals. The sections were reassembled and then prepared to size 50 Master Apical Rotary and size 40 Master Apical Files, for the Lightspeed and K file groups, respectively. Similar 35-mm slides of the instrumented canals were taken and electronically superimposed over their uninstrumented counterparts. Displacements of root canal centers after preparation and increases in cross sectional areas of the prepared root canals were evaluated at specific levels. No significant differences in displacement of canal centers or between the mean cross-sectional areas of the instrumented root canals were recorded between the Lightspeed and K-file groups. Both types of instruments produced similar displacement of canal centers after preparation. PMID- 11487152 TI - In vitro infrared thermographic assessment of root surface temperatures generated by the thermafil plus system. AB - The purpose of this in vitro study was to investigate root surface temperatures generated by the Thermafil Plus system using infrared thermography. A total of 60 extracted human maxillary central incisors and maxillary first molars were biomechanically instrumented using ProFile 0.04 Taper Series 29 rotary instruments. The root canals were obturated using the Thermafil Plus system. An infrared thermography camera was used to measure the external root surface temperatures during and after the completion of obturation. The mean temperature rise of the external root surface from the ambient temperature was 4.26 degrees C for the mesial buccal roots, 4.58 degrees C for the distobuccal roots, 4.76 degrees C for the palatal roots, and 4.87 degrees C for the anterior roots. These temperatures were well below the critical level of 10 degrees C needed to cause damage to the attachment apparatus. From this in vitro experiment, no evidence was discovered which would indicate that the Thermafil Plus obturation system would result in damage to the periodontal ligament and the surrounding attachment apparatus as a consequence of temperature rise on the external root surface. PMID- 11487153 TI - Effectiveness of three endodontic irrigants at various tubular depths in human dentin. AB - Bacteria from infected root canals can invade dentinal tubules, thus dentin disinfection is an important aspect of endodontic therapy. This study compares three endodontic irrigants for efficiency in killing bacteria established within human dentinal tubules. Root canals in extracted teeth were prepared and sterilized. Broth cultures of Enterococcus faecalis were allowed to grow within the canals to penetrate dentinal tubules. The infected canals were exposed individually to each of the irrigants for 1 min. Irrigants were 0.525% sodium hypochlorite, Tubulicid (0.2% EDTA), and 0.12% chlorhexidine (Peridex). Sterile water was the control. Viable bacteria were analyzed by drilling incrementally into dentin from the cementum toward the canal. Smaller diameter drills were used for each depth. Shavings were cultured at three depths, for each of three root levels: coronal, midroot, and apical. Although considerable variation occurred between roots, sodium hypochlorite seemed to be superior. Tubulicid and Peridex were better than water. More bacteria remained viable at greater distances from the pulp. These observations apparently apply to all levels in the canal. PMID- 11487154 TI - An in vitro evaluation of the accuracy of the root ZX in the presence of various irrigants. AB - The purpose of this study was to evaluate the accuracy of the Root ZX in vitro in the presence of a variety of endodontic irrigants. The in vitro model, described by Donnelly, consisted of refrigerated gelatin made with 0.9% sodium chloride instead of water. The following irrigants were tested: 2% lidocaine with 1:100,000 epinephrine, 5.25% sodium hypochlorite, RC Prep, liquid EDTA, 3% hydrogen peroxide, and Peridex. A total of 30 extracted, single-rooted teeth were used. The experimental measurements in the presence of the various irrigants were compared with the actual canal lengths. The present data indicate that the Root ZX electronic apex locator reliably measured canal lengths to within 0.31 mm and that there was virtually no difference in the length determination as a function of the seven irrigants used. These results strongly support the concept that the Root ZX is a useful, versatile, and accurate device for the determination of canal lengths over a wide range of irrigants commonly used in the practice of endodontics. PMID- 11487155 TI - Wear resistance of cryogenically treated stainless steel files. AB - Cryogenic treatment, which involves ultra-sub-zero treatment of metal alloys, has been shown to improve the wear resistance of several types of stainless steel. The purpose of this study was to examine the effect of cryogenic treatment on wear resistance of Flex-R and Hedstrom (Union Broach) files. These instruments were attached to an Instron testing machine and underwent 300 push-pull strokes (6-mm movement range, 600-mm/min speed, 1-N loading) against 1.5-mm thick dentin wafers. Their wear was determined by comparing the depth of grooves (without changing file position) cut in acrylic specimens (1.5-mm thick) before and after machining dentin. The mean and standard deviation (n = 20) of the relative change in cutting efficiency was determined for the files. An analysis of variance showed that the cutting efficiency of all files decreased significantly after wearing on dentin (p < 0.0001). However, when comparing the post-/pre-cutting efficiency ratios of the files, it was seen that cryogenic treatment did not affect the wear resistance of the files. PMID- 11487156 TI - Changes in root canal geometry after preparation assessed by high-resolution computed tomography. AB - Root canal morphology changes during canal preparation, and these changes may vary depending on the technique used. Such changes have been studied in vitro by measuring cross-sections of canals before and after preparation. This current study used nondestructive high-resolution scanning tomography to assess changes in the canals' paths after preparation. A microcomputed tomography scanner (cubic resolution 34 microm) was used to analyze 18 canals in 6 extracted maxillary molars. Canals were scanned before and after preparation using either K-Files, Lightspeed, or ProFile .04 rotary instruments. A special mounting device enabled precise repositioning and scanning of the specimens after preparation. Differences in surface area (deltaA in mm2) and volume (deltaV in mm3) of each canal before and after preparation were calculated using custom-made software. deltaV ranged from 0.64 to 2.86, with a mean of 1.61 +/- 0.7, whereas deltaA varied from 0.72 to 9.66, with a mean of 4.16 +/- 2.63. Mean deltaV and deltaA for the K-File, ProFile, and Lightspeed groups were 1.28 +/- 0.57 and 2.58 +/- 1.83; 1.79 +/- 0.66 and 4.86 +/- 2.53; and 1.81 +/- 0.57 and 5.31 +/- 2.98, respectively. Canal anatomy and the effects of preparation were further analyzed using the Structure Model Index and the Transportation of Centers of Mass. Under the conditions of this study variations in canal geometry before preparation had more influence on the changes during preparation than the techniques themselves. Consequently studies comparing the effects of root canal instruments on canal anatomy should also consider details of the preoperative canal geometry. PMID- 11487157 TI - Effect of ultrasonic vibration and various sealer and cement combinations on titanium post removal. AB - The purpose of this study was to determine the effect of ultrasonic vibration on the force required to remove prefabricated posts. Ninety-six extracted human canines were divided into eight groups, which were prepared and obturated with gutta-percha and either a eugenol-containing (Roth's 801 Elite) or eugenol-free (AH26) sealer. Titanium #6 Parapost XH posts were cemented with either zinc phosphate or Panavia 21 resin cement. One half of the sample was subjected to ultrasonic vibration for 16 min at the post-dentin interface, whereas the other half received no vibration (controls). Each combination of sealer, cement, and vibration status was subjected to tensile load to failure using an Instron testing machine. Posts cemented in teeth obturated with gutta-percha and AH26 sealer demonstrated significantly greater resistance to dislodgement, compared with teeth obturated with gutta-percha and Roth's 801 Elite sealer. There was no statistical difference in retention of posts cemented with either zinc phosphate or Panavia 21, regardless of the sealer used. Additionally ultrasonic vibration increased post retention for both cements. PMID- 11487158 TI - Evaluation of fiber-composite laminate in the restoration of immature, nonvital maxillary central incisors. AB - The purpose of this study was to evaluate the use of fiber-composite laminate, a reinforcement ribbon embedded throughout a composite restoration, to reinforce immature maxillary central incisors. Twenty-six mature maxillary central incisors were prepared a minimum of 3 mm below the facial cementoenamel junction to simulate immature nonvital teeth. They were separated into 3 groups: group 1 was unfilled and served as a control; group 2 was filled to the depth of the preparation with composite; and group 3 was filled with composite and Connect Reinforcement Ribbon. The specimens were subjected to class I loading in an Instron Testing Machine until catastrophic failure occurred. The results indicate a highly significant difference between the groups (p < or = 0.003). Group 1 fractured at an average load of 31.08 kg, group 2 at 51.00 kg, and group 3 at 37.93 kg. These findings suggest that composite alone increases fracture resistance of the immature tooth model more than composite with Reinforcement Ribbon. PMID- 11487159 TI - Permeability characteristics of bovine and human dentin under different pretreatment conditions. AB - In order to use bovine dentin instead of human dentin for in vitro adhesion and cytotoxicity tests the permeability characteristics of human and bovine dentin should be similar. In the present study hydraulic conductance (Lp) and diffusional water flux (J5) of human and bovine dentin slices were compared. The permeability experiments were performed in a split chamber using tritiated water in physiological saline. Lp and Js of bovine dentin were 0.7- to 2.4-fold and 1.1 to 3.5-fold that of human dentin (not statistically significant). For human and bovine dentin Lp and Js increased with etching and showed an inverse linear relationship (r > or = 0.7) with dentin thickness. The variability of bovine data was low (perfusion = 30%, diffusion = 22%) and about half that of the human data. In conclusion bovine dentin near the cementoenamel junction seems to be a suitable alternative for coronal human dentin for in vitro tests with respect to transdentinal permeability characteristics. PMID- 11487160 TI - Determination of lidocaine in dental pulp by high-performance liquid chromatography. AB - The purpose of this study was to develop a method for lidocaine detection in dental pulp by high-performance liquid chromatography. The amounts of lidocaine in dog pulps were quantitated after local injection to evaluate lidocaine recovery from pulp tissue with this technique. Comparison was also made between the amount of lidocaine found in upper and lower canines. The high-performance liquid chromatography system was shown to be a reliable and reproducible tool for lidocaine determination. Lidocaine extraction from the tissue showed recovery of 90%. The amount of lidocaine recovered from the upper canine (0.21 microg/mg) was higher than the lower canine (0.17 microg/mg). PMID- 11487161 TI - Bacterial coronal leakage after obturation with three root canal sealers. AB - The purpose of this study was to compare the bacterial leakage of root canals obturated with three root canal sealers, using Endodontalis faecalis as a microbial tracer to determine the length of time for bacteria to penetrate through the obturated root canal to the root apex. Seventy-five, single-rooted teeth with straight root canals had the crown cut off at the cementoenamel junction. Root canals were instrumented by a step-back technique. The prepared teeth were randomly divided into 3 groups of 19 teeth each and another 2 groups as positive and negative controls (9 teeth each). The experimental groups were dependent on the sealer used: AH-Plus, Apexit, and Ketac-Endo. The root canals were obturated using a lateral condensation technique. After 24 h the teeth were attached to microcentrifuge tubes with 2 mm of the root apex submerged in Brain Heart Infusion broth in glass test tubes. The coronal portions of the root canal filling materials were placed in contact with E. faecalis. The teeth were observed for bacterial leakage daily for 30 and 60 days. With the chi2 test for comparing pairs of groups at the 0.05 level (p < 0.05), there was no statistical difference between Ketac-Endo and AH-Plus (p > 0.06), but Apexit had significantly higher leakage (p < 0.05) at 30 days. After 60 days there was no statistical difference between Ketac-Endo and Apexit (p > 0.05), but Apexit leaked more than AH-Plus. The conclusion drawn from this experiment was that epoxy resin root canal sealer was found to be more adaptable to the root canal wall and filling material than a calcium hydroxide sealer when bacterial coronal leakage was studied. PMID- 11487162 TI - A comparative evaluation of three root-end filling materials: an in vitro leakage study using Prevotella nigrescens. AB - This study used Prevotella nigrescens to evaluate the sealing ability of Geristore, Super-EBA, and ProRoot when used as root-end filling materials. One hundred single-canal, extracted, human teeth were cleaned and shaped with Gates Glidden burs and .04 Taper (ProFile Series 29) rotary instruments. The root-ends were resected, and a 3-mm deep root-end preparation was made with ultrasonic tips. After steam sterilization, the teeth were randomly divided into three groups of 30. Each group was root-end-filled with a different material. Five root end cavities were left open and served as positive controls; another five root end cavities filled with sticky wax and covered with two layers of nail polish acted as negative controls. The teeth were attached to presterilized snap-on plastic caps; the root-ends submerged into 12-ml vials of chopped meat carbohydrate broth and placed in an anaerobic chamber. Twice a week, a sterile micropipette was used to inoculate 0.1 ml of a broth culture of Prevotella nigrescens into the root canal of each tooth. Results after 47 days indicated there were no significant differences between the three root-end filling materials against penetration of Prevotella nigrescens. PMID- 11487163 TI - PH changes at the surface of root dentin after intracanal dressing with calcium oxide and calcium hydroxide. AB - This study was designed to compare pH changes at the cervical and apical surfaces of root dentin after canal obturation with calcium oxide or calcium hydroxide pastes. The effect of the exposure to CO2 on the pH at the root surface also was assessed for both materials. Thirty-six extracted human canines were selected and instrumented. Wells 1 mm deep x 1.5 mm in diameter were drilled in the cervical and apical regions of the buccal root surface. The teeth were randomly divided into two groups and obturated with either calcium hydroxide or calcium oxide. pH was measured in the cervical and apical wells at varying time intervals until 48 days posttreatment. After pH measurement on day 48 the vials were flushed with CO2 and the pH measured again at days 53 and 68. The results indicated a similar pattern of pH increase after treatment with either calcium oxide or calcium hydroxide. pH dropped significantly after exposure to CO2 in both groups. This study demonstrated that hydroxyl ions produced when calcium oxide reacts with water diffuse through the dentinal tubules to the surface of the root in a manner similar to hydroxyl ions from calcium hydroxide. PMID- 11487164 TI - An evaluation of endodontically treated vertical root fractured teeth: impact of operative procedures. AB - Vertical root fractures of endodontically treated teeth are a frustrating complication that leads to extraction. The aim of the current survey was to evaluate the role of operative procedures in the etiology of this complication. A total of 154 endodontically treated vertical root fractured teeth were cleaned and washed after extraction and maintained in individual vials. Periapical radiographs before extraction, clinical findings and previous operative procedures were recorded. A post was observed in 95 teeth (61.7%), with 66 of these ending at the coronal third of the root. Most were screw posts of the Dentatus type (n = 64) and tapered cast posts (n = 14). A full crown was observed in 118 teeth, and 65 of these (55%) were extracted between 1 to 5 yr after final restoration. In 24 crowned teeth extraction was conducted within 1 yr after restoration and in 28 teeth after >5 years. It was concluded that post placement and root canal treatment are the major etiological factors for root fractures. Because signs and symptoms can appear years after the operative procedures in the root have been completed, coronal restorations would not interfere with the correct clinical diagnosis of vertical root fractures. Frequent recalls are recommended to diagnose vertical root fractures early, especially in susceptible teeth, such as premolars and mesial roots of mandibular molars. PMID- 11487165 TI - Frequency of overinstrumentation with an acceptable radiographic working length. AB - The aim of this in vitro study was to determine how frequent a seemingly accurate working length ending radiographically 0 to 2 mm short of the radiographic apex resulted in an instrumentation beyond the apical foramen. Under simulated clinical conditions working lengths of 169 root canals were radiographically determined in 91 extracted teeth. In all cases the measuring files adjusted to the final working length (Iwork) were located 0 to 2 mm short of the radiographic apex. Iwork was subsequently compared with the actual reference length (Iref) representing the distance between the apical foramen and the coronal reference. Instrumentation beyond the apical foramen (Iwork > ref) occurred in premolars in 51% (95% confidence interval: 36%; 66%) of the cases, in molars in 22% (95% confidence interval: 14%; 30%), and in anterior teeth in no case. These results suggest that in premolars and molars a radiographically working length ending 0 to 2 mm short of the radiographic apex provides, more often than expected, a basis for unintentional overinstrumentation. PMID- 11487166 TI - Effect of prophylactic amoxicillin on endodontic flare-up in asymptomatic, necrotic teeth. AB - The purpose of this prospective, randomized, double-blind, placebo-controlled study was to determine the effect of prophylactic amoxicillin on the occurrence of endodontic flare-up in asymptomatic, necrotic teeth. Seventy patients participated and had a clinical diagnosis of an asymptomatic, necrotic tooth with associated periapical radiolucency. One hour before endodontic treatment, patients randomly received either 3 g of amoxicillin or 3 g of a placebo control in a double-blind manner. After endodontic treatment, each patient received: ibuprofen; acetaminophen with codeine (30 mg); and a 5 1/2-day diary to record pain, swelling, percussion pain, and number and type of pain medication taken. The results demonstrated 10% of the 70 patients had a flare-up characterized by moderate-to-severe postoperative pain or swelling that began approximately 30 h after endodontic treatment and persisted for an average of 74 h. Of the seven patients who had flare-ups, 4 were in the amoxicillin group and 3 were not. Prophylactic amoxicillin did not significantly (p = 0.80) influence the endodontic flare-up. We concluded that a prophylactic dose of amoxicillin before endodontic treatment of asymptomatic, necrotic teeth had no effect on the endodontic flare-up. PMID- 11487168 TI - Bur extensor for root canal access. AB - Access to a tooth with a partial canal calcification was made. The access was possible only with a bur extensor connected to a LN bur. Calcification was bypassed, and the canal was completely prepared. PMID- 11487167 TI - Comparison of nickel-titanium file distortion using electric and air-driven handpieces. AB - The use of rotary instrumentation has increased dramatically in endodontics. Air driven and electric handpieces are both currently available for use with the nickel-titanium files. To date no studies have compared the frequency of instrument separation and/or distortion while using the two types of handpieces with rotary files. The purpose of this study was to determine whether there is a difference in the frequency of file separation/distortion when an electric handpiece is compared with an air-driven handpiece. Extracted human molar teeth were instrumented with nickel-titanium Profile ISO .04 taper rotary files (Dentsply Tulsa Dental Products, Tulsa, OK) under controlled conditions and the results examined for statistical significance. The teeth chosen for the study fit the following criteria: minimal caries, curvature of any root no greater than 60 degrees and at least one root with a minimum of 30-degree curvature. The air driven handpiece was used at 167.67 rpm and the electric handpiece at 150 rpm per manufacturer's recommendations. Files were examined both before and after use to evaluate whether breakage or deformation had occurred. There was no significant difference in file breakage or distortion when the handpieces were compared. There were, however, a significantly greater number of file separations/ distortions noted in the #8 Profiles (ISO size 20) than with the other files. It may be prudent to view this file as a disposable instrument. PMID- 11487169 TI - Effect of an acid environment on leakage of root-end filling materials. AB - One hundred fifty-six single-rooted human extracted teeth received root canal treatment, apicoectomy, and ultrasonic root-end preparations. The roots were divided into six groups of 24 teeth. The groups were filled with Amalgam, Geristore, Super-EBA, Mineral Trioxide Aggregate (MTA), Calcium Phosphate Cement (CPC), or MTA with CPC matrix, respectively. Immediately after root-end filling, 12 teeth from each group were exposed to a pH of 5.0 for 24 h, and 12 teeth were exposed to a pH of 7.4 for 24 h. Twelve teeth served as controls. All teeth were exposed to Pelikan Ink for 5 days and cleared. Linear dye leakage was recorded. Data were statistically analyzed. An acid pH significantly reduced dye leakage of Geristore (0.67 vs. 3.93 mm) and MTA with CPC matrix (0.54 vs. 2.41 mm), whereas leakage of all other materials was not affected by pH. In conclusion an acid environment did not hinder the sealing ability of any of the materials tested, and enhanced the sealing ability of Geristore and MTA with CPC matrix. PMID- 11487170 TI - Effects of erbium,chromium:YSGG laser irradiation on root canal walls: a scanning electron microscopic and thermographic study. AB - The present study evaluates the morphological changes in root canal walls and temperature changes at root surfaces as a result of intracanal irradiation by erbium,chromium:YSGG laser under various conditions in vitro. Sixty single-rooted human teeth were examined. Root canals were prepared, and laser irradiation was performed using an optic fiber at output powers ranging from 1 to 6 W with or without water spray cooling. Specimens were evaluated by stereoscopy, scanning electron microscopy, and thermography. Carbonization and cracks were observed in all samples irradiated without cooling, whereas little or no carbonization and no smear layer or debris were observed in samples irradiated with cooling. Maximum temperature rise at irradiation without cooling was above 37 degrees C, whereas that at irradiation with cooling was 8 degrees C. Results of the present study indicate that erbium,chromium:YSGG laser irradiation with water spray cooling is a useful method for removal of smear layer and debris from root canals. PMID- 11487171 TI - Indirect calorimetry and nutritional problems in clinical practice. AB - Indirect calorimetry is a simple and affordable tool for measuring energy expenditure and for quantifying the utilization of macronutrients. Its use is becoming increasingly widespread, but it is necessary to know its methodological features and its theoretical and practical limitations. Indirect calorimetry measures the rate of resting energy expenditure (REE), the major component of the total daily energy expenditure. Thus, indirect calorimetry reliably estimates the individual energy needs. Coupling the measurement of body composition to that of REE expands the diagnostic potential of indirect calorimetry. Once the lean and fat compartments have been measured, it is possible to establish on the basis of REE whether an individual is hyper- or hypometabolic. The evaluation of substrate oxidation by indirect calorimetry is subject to more severe theoretical constraints, because certain metabolic assumptions must be made. The clinical applications are practically unlimited. In the critically ill, a major goal is to maintain energy balance during the hypermetabolic response following trauma. The REE measurement is valuable from the diagnostic standpoint, because it recognizes discrepancies from the expected time-course of hypermetabolism, for example signaling a potentially catastrophic hypometabolic response. REE is also indispensable for providing correct nutritional support because both hyper- and undernutrition lead to increased mortality. In young or elderly patients, in whom energy consumption may be very different from that predicted from equations based on anthropometric measures, indirect calorimetry is particularly useful. PMID- 11487172 TI - Some applications of indirect calorimetry to sports medicine. AB - Some applications of indirect calorimetry to sports medicine are discussed and exemplified by case reports. In particular, it is suggested that oxigen consumption can be employed to assess the effects of physical activity on fat free tissues and that the respiratory quotient may offer some insights into the food habits of athletes. PMID- 11487173 TI - The use of REE determination in a clinical setting applied to respiratory disease. AB - The determination of a patient's nutritional condition is an important moment of clinical evaluation in everyday experience. Quite often, it is important to identify symptoms, signs and objective parameters that may reveal a malnourished condition. To satisfy this request, clinicians recur to clinical examination and biochemical tests. It is also possible to use indirect calorimetry. Through the evaluation of oxygen consumption (VO2) and carbon dioxide production (VCO2), indirect calorimetry permits the respiratory quotient (RQ) to be calculated and the exact energy consumption during resting conditions to be determined. This technology, currently used in research departments and in hospital units, is more accurate for the estimation of resting energy expenditure (REE) than the Harris Benedict equation, especially in the case of patients affected by respiratory diseases or other systemic conditions. The evaluation of REE through indirect calorimetry is a useful monitoring and prognostic index in acutely and chronically ill patients. PMID- 11487174 TI - Influence of age on clinical and immunological characteristics of newly diagnosed type 1 diabetic patients. AB - The aim of this study was to analyse the immunological and clinical characteristics of a group of patients at the onset of type 1 diabetes and to determine if these findings are age related. For this purpose, 68 newly diagnosed type 1 diabetes mellitus patients referred to our hospital between 1997 and 1999 were studied; 42 were adults (mean age 24+/-3.5 years) and 26 children (mean age 6.1+/-4 years). Autoantibody markers islet cell antibodies, glutamic acid decarboxylase antibodies (GADA) and tyrosine phosphatase antibodies (IA-2A), pancreatic reserve (glucagon test) and HbA1c were determined. Some clinical characteristics, such as mode of presentation and insulin requirements, were also analysed. Type 1 diabetes mellitus was found to be autoimmune in 83.8% of the patients and idiopathic in 16.2%, without significant differences between adults and children. In the whole autoimmune group, GADA was more prevalent in adults and IA-2A more frequent in children. On the other hand, adults showing autoimmune markers developed ketosis more frequently and needed higher insulin doses at diagnosis, while children did not exhibit clinically significant differences associated with the presence or absence of antibodies. In conclusion, in children the presence of autoimmune markers is not related to the mode of presentation or characteristics of type 1 diabetes. In adults, however, the autoimmune group presents with more-severe clinical disease than antibody negative patients. Age at onset seems to be an important parameter in the natural history of type 1 diabetes and must be taken into account in epidemiological or intervention studies. PMID- 11487175 TI - Cholesteryl ester transfer and cholesterol esterification in type 1 diabetes: relationships with plasma glucose. AB - The activities of two crucial enzymes of reverse cholesterol transport, cholesterol ester transfer protein (CETP) and lecithin:cholesterol acyltransferase (LCAT), and their relationships with lipid profile and fasting plasma glucose were examined in 35 type 1 diabetic children. The CETP and LCAT activities were significantly lower (p<0.05) in the 4 subjects with normal fasting plasma glucose levels (<6.39 mmol/l) than in the 28 with high plasma glucose levels (CEPT activity, 10.63+/-3.81 vs. 32.18+/-13.94 nmol/ml h; LCAT activity, 25.52+/-4.53 vs. 39.52+/-12.52 nmol/ml h; both p<0.05). The subjects with high plasma glucose levels also had higher total and LDL-cholesterol than those with normal glucose levels. CETP activity was positively correlated with fasting plasma glucose, CETP concentration, LCAT activity, total cholesterol, free cholesterol, LDL-C, and LDL-cholesteryl ester, while negatively correlated with cholesteryl ester to free cholesterol ratio, LDL triglyceride to protein ratio, and LDL triglyceride to cholesteryl ester ratio. LCAT activity was found to positively correlate with CETP activity, total cholesterol, free cholesterol, LDL-C, CETP concentration, and LDL-cholesteryl ester, while it negatively correlated with cholesteryl ester to free cholesterol ratio. The results observed in type 1 diabetic subjects suggest that (1) accelerated LCAT and CETP activities may result in the accumulation of LDL-cholesteryl ester; and (2) fasting plasma glucose may be a major determinant of CETP activity. PMID- 11487177 TI - Evaluation of the efficacy of the combination of multiple autoantibodies to islet specific antigens in Korean type 1 diabetic patients. AB - With the recent cloning and recombinant expression of novel islet autoantigens, it is possible to investigate whether the quantitative expression of autoantibodies to these molecules is correlated with the semiquantitative titration of islet cell antibodies (ICA). To estimate the regional difference in the degree of association between these autoantibodies, autoantibodies reacting with ICA512/IA-2 (ICA512) and GAD65 in addition to ICA were analyzed in 131 Korean children with type 1 diabetes. Among the 131 type 1 diabetes patients, 36% was ICA512-positive, 56% was anti-GAD65-positive, and 43% was ICA-positive. However, in a subset of these with recent onset (<1 year), the prevalences of ICA512, anti-GAD65, and ICA were 75%, 75%, and 87.5%, respectively. ICA512 as well as anti-GAD65 were significantly associated with the presence of ICA. Among type 1 diabetes patients, 69% had one or more and 43% had two or more of these autoantibodies. Autoantibodies to ICA512 and anti-GAD65 were observed in 40 and in 41 of 56 ICA-positive subjects, respectively. Furthermore, ICA512 or anti GAD65 were positive in 97% (34 of 35) and 100% (22 of 22) of patients with ICA levels > or =20 JDF-u and > or =40 JDF-u, respectively. The titer of ICA correlated with those of ICA512 (r=0.41, p<0.001) and anti-GAD65 (r=0.49, p<0.001). Both the prevalences and the titers of ICA512 or anti-GAD65 were strongly correlated with those of ICA even in Korean type 1 diabetes patients. ICA512 and anti-GAD65 in combination may be considered to be an alternative to ICA. PMID- 11487176 TI - The state of leukocyte adhesiveness/aggregation in the peripheral blood of patients with type 2 diabetes and ischemic vascular disease. AB - White blood cells have a potential role in the pathogenesis of vasculopathy in diabetic patients. We studied the circulating peripheral blood in a cohort of patients with documented ischemic heart or brain disease with and without type 2 diabetes by means of image analysis and flow cytometry. Our study showed that the state of leukocyte adhesiveness/aggregation is slightly increased in those who had concomitant diabetes but that there was no difference regarding the expression of CD11b/CD18 and CD62L antigens on the surface of the peripheral blood white blood cells. The finding of a significantly increased number of white blood cells in the peripheral blood of patients with ischemic vascular diseases is important insofar as it is associated with a poorer prognosis. PMID- 11487178 TI - Pathogenesis of obesity and diabetes mellitus: insights provided by indirect calorimetry in humans. AB - Energy homeostasis is the balance between energy intake and energy expenditure. Assessment of energy intake is unreliable, especially in obese individuals. On the contrary it is possible to assess energy expenditure by means of different techniques. In this review, the contributions of indirect calorimetry to its assessment and to the comprehension of the pathogenic mechanisms of obesity and diabetes mellitus are discussed. Knowledge regarding energy expenditure and its components (basal metabolic rate, food-induced thermogenesis, expenditure related to physical activity), obtained in obese individuals by means of indirect calorimetry, is summarized. The effects of dietetic manipulation of the relative fractions of macronutrients (fat, carbohydrate and protein) and fiber intake on energy expenditure are described with implications regarding the pathogenesis of obesity. Besides environmental factors, energy expenditure is strongly influenced by inherited features. The application of indirect calorimetry to study recently discovered polymorphisms of genes strongly related to energy expenditure in humans is also summarized. The role of indirect calorimetry in the identification of the glucose metabolic pathways (oxidative and non-oxidative) affected in the insulin resistance syndrome is also described. Indirect calorimetry has been useful in realizing that the alterations typical of insulin resistance are reproducible in vivo in healthy humans, increasing the availability of free fatty acids. In fact, indirect calorimetry contributed to understanding the in vivo mechanisms of substrate competition, which was hypothesized more than forty years ago. In this field of application, the identification of simple (non-invasive and inexpensive) markers of insulin resistance is urgent for primary and secondary prevention of these diseases with high morbidity and mortality. The postabsorptive assessment of resting energy expenditure and macronutrient partitioning in fuel metabolism may be helpful and is discussed with advantages and limitations. The treatment of multifactorial diseases will probably require the use of poly-therapeutic interventions. In fact, drugs that act purely on one pathogenic factor of these diseases frequently fail. To better design strategic therapeutic approaches for each patient, to achieve significant long-term efficacy in the treatment of these diseases and to prevent their complications, indirect calorimetry has to be considered a landmark of quality control for nutritional support. PMID- 11487179 TI - Functional mapping of cortical areas. PMID- 11487181 TI - Unawareness of dyskinesias in Parkinson's and Huntington's diseases. AB - We performed a clinical study to evaluate the unawareness of dyskinesias in patients affected by Parkinson's disease (PD) and Huntington's disease (HD). Thirteen PD patients with levodopa-induced dyskinesias and 9 HD patients were enrolled. Patients were asked to evaluate the presence of dyskinesias while performing specific motor tasks. The Abnormal Involuntary Movement Scale (AIMS) and Goetz dyskinesia rating scale were administered to determine the severity of dyskinesias. The Unified Parkinson's disease rating scale (UPDRS) and Unified Huntington's Disease Rating Scale (UHDRS) were used in PD and HD patients, respectively. In PD we found a significant negative relationship between unawareness score at standing and AIMS score and between unawareness score at hand pronation-supination and AIMS score for upper limbs. In HD we found a significant positive relationship between total unawareness score and UHDRS score and between total unawareness score and disease duration. In PD the unawareness seems to be inversely related with severity of dyskinesias, while in HD it is directly related to disease duration and severity. PMID- 11487180 TI - Epidemiology of progressive supranuclear palsy. ESGAP Consortium. European Study Group on Atypical Parkinsonisms. AB - Progressive supranuclear palsy (PSP) is a rare form of parkinsonism. The incidence rates are about 0.3-1.1 cases per 100,000 persons. The only two case control studies performed up to now show conflictual results as regards education and residence in rural areas. Recently, a cluster of PSP and atypical parkinsonism has been observed in French Antilles. The hypothesis is that a consumption of both tropical fruit and herbal tea may be associated with PSP onset. Some PSP families with a probably autosomal dominant transmission have been described. A high frequency of a tau haplotype (H1/H1) associated with PSP is reported by some authors. The significance of this association is still not clear. We have performed a case-control study on 58 PSP cases, 116 hospital controls and 58 population controls. PMID- 11487182 TI - A PET study with [11-C]raclopride in Parkinson's disease: preliminary results on the effect of amantadine on the dopaminergic system. AB - Amantadine has been proved to be beneficial in Parkinson's disease. Although it is still uncertain which neurochemical events are modified at therapeutic doses, an increase in dopaminergic tone secondary to NMDA receptor blockade and a direct inhibition of the glutamatergic overactivity have been suggested to be involved in its clinical effects. The aim of this study was to evaluate the effects of amantadine on the dopaminergic system by measuring the in vivo binding of [11 C]raclopride to D2 dopamine receptors in the basal ganglia of 6 patients with idiopathic Parkinson's disease. Each patient underwent a PET study, before and after 14 days of treatment with amantadine (200 mg/day). Repeated treatment with therapeutic doses of amantadine induced a moderate increase in the in vivo binding of [11C]raclopride in the putamen of PD patients. This observation indicates that in PD patients, 200 mg/day amantadine does not produce an increase in extracellular levels of dopamine sufficiently to inhibit raclopride binding or that, if present, is it masked by a concurrent increase in receptor availability, as recently reported in rat striatum. PMID- 11487183 TI - Efficacy and tolerability of dopamine agonists in a parkinsonian population. AB - A clinical retrospective study was carried out in a population of 366 Parkinson's disease (PD) outpatients, to analyse the efficacy and tolerability of nonergoline and ergoline dopamine agonist (DA), in monotherapy or in combination with L-dopa. Safety was comparable in both groups except for higher occurrence of gastrointestinal symptoms in ergoline group and somnolence in nonergoline group. No significant difference concerning efficacy and tolerability was found during DA monotherapy. Mean age at PD onset was slightly higher in patients withdrawing DA monotherapy for adverse events comparing to patients who needed the addition of L-dopa (60.36 +/- 7.53 versus 54.88 +/- 10.75; p<0.05), suggesting that older age at the onset of the disease increases the risk for adverse events during DA monotherapy. The follow-up of the remaining patients still in monotherapy with DA will allow a better evaluation of these aspects. PMID- 11487184 TI - Neurophysiological classification of ulnar entrapment across the elbow. AB - Ulnar nerve entrapment across the elbow (UAE), a common entrapment, requires neurophysiological evaluation for a diagnosis, but a standardised neurophysiological classification is not available. The aim of our study was to evaluate the validity of a neurophysiological classification of UAE, developed by us. To this end, we examined whether sensorimotor deficits, as observed by the physician and as referred by the patients, increased with the neurophysiological severity according to the classification. We performed a multiperspective assessment of 63 consecutive arms from 52 patients with a clinical diagnosis of UAE. Neurophysiological, clinical and patient-oriented validated measurements were used. The neurophysiological classification is based on the presence or absence of evoked responses and on the normality or abnormality of conduction findings. A strict relationship was observed between the degree of neurophysiological severity and the clinical findings (sensorimotor deficits). Moreover, a significant positive correlation between hand functional deficit and neurophysiological classification was observed. Conversely, a clear correlation between neurophysiological pattern and symptoms was not found. The neurophysiological classification is easy to use and reliable, but further multicentric studies should be performed. PMID- 11487185 TI - Physiologic study of the subthalamic volume. AB - Deep brain stimulation (DBS) obtains good control of advanced PD symptoms. Chronic stimulation of Stn may alleviate rigidity, dyskinesia and tremor. Anatomical and functional intraoperative mapping are mandatory to obtain careful target localisation. Per-operative macrostimulation was carried out in 22 patients undergoing bilateral DBS in Stn; a volume 6 mm above to 4 mm below Stn was explored. Positive, collateral and adverse effects were recorded every 2 mm. Results obtained during acute stimulation were correlated to anatomical data from stereotactic atlases. Our findings suggest a volume, encompassing the zona incerta, Forel's fields and the lowermost part of anterior thalamus, functionally homogeneous to Stn. In fact, the stimulation of this volume obtains reduction of PD symptoms comparable to Stn. PMID- 11487186 TI - Reversal of CSF positivity for JC virus genome by cidofovir in a patient with systemic lupus erythematosus and progressive multifocal leukoencephalopathy. AB - We report the case of a 36-year-old woman affected by systemic lupus erythematosus who developed rapidly progressive multifocal leukoencephalopathy. Cidofovir therapy induced disappearance of JC virus genome from the cerebrospinal fluid and stabilization of the MRI picture. Despite the fatal outcome after a few months of disease, cidofovir treatment deserves further testing as single antiviral therapy in HIV-negative PML patients. PMID- 11487187 TI - Adult-onset adrenoleukodystrophy: a clinical and neuropsychological study. AB - Adrenoleukodystrophy (ALD) is a rare, X-linked inherited metabolic disease, which is associated with the accumulation of very long chain fatty acids (VLCFA) in the adrenal glands and in the central and peripheral nervous systems. ALD leads to leukoencephalic myeloneuropathy and hypoadrenalism. The two most frequent phenotypes are infantile ALD and adrenomyeloneuropathy (AMN), which account for 80% of cases. Adult-onset ALD is rare (1-3%). The cases reported so far have had a variable course and have often been characterised by prevalently psychiatric symptoms. We describe the case of a 37-year-old man affected by ALD whose symptoms first appeared a few years earlier in the form of worsening personality and memory disturbances, and were followed by the onset of spastic paraparesis and amyotrophy indicative of the concomitant presence of AMN. The neuropsychological alterations configure a picture of mainly frontal cortico subcortical dementia, which is supported by magnetic resonance findings. PMID- 11487188 TI - Segmental myoclonus in a patient affected by syringomyelia. AB - We describe a patient who has been complaining of brief jerk-like, rhythmic, involuntary movements involving the second digit of the left hand for the last three months. These involuntary jerks produced an adduction movement of the second digit and were unaffected by peripheral sensory stimuli. In addition, the patient experienced loss of dexterity in the left hand. On examination the patient showed hypotrophy of the first dorsal interosseous (FDI) muscle of the left hand and a dissociated sensory loss involving the C8-T1 dermatomes. Magnetic resonance imaging of the brain and spinal cord revealed a tonsilar hemiation characteristic of the Chiari I malformation associated with a syrinx extending from C4 to D5 that did not communicate with the fourth ventricle. The electrophysiological evaluation indicated the presence of a focal myoclonus of spinal origin that is likely to be caused by the syrinx. PMID- 11487189 TI - The value of cortical stimulation applied to the surgery of malignant gliomas in language areas. AB - This study analyzes the utility of peroperative cortical language mapping applied to the surgery of high-grade gliomas situated within or in close vicinity to speech areas. Fifteen consecutive patients harboring high-grade gliomas located in the dominant hemisphere, causing regressive or minor language troubles, underwent awake craniotomy in our institution between June 1998 and April 2000. The technique of cortical stimulation under local anesthesia for language mapping, initially described by Ojemann and colleagues, was applied with some modifications. All patients tolerated awake craniotomy except one, who was intubated after the mapping procedure. Mapping results confirmed a high variability in location of language sites. It was possible to achieve a gross total tumor removal in all cases. Nine patients (60%) exhibited a transient postoperative aggravation. Two patients (13%) presented permanent phasic aggravation. One patient died 16 days after surgery from pulmonary embolism. Five patients died for tumor progression, with a mean survival time of 16.4 months and a median high-quality survival period of 14.2 months. With a mean follow-up of 9.9 months (range, 18-6 months), the 9 survivors are recurrence-free and reveal no significant change in linguistic abilities. This technique is well tolerated and consents to maximize the extent of surgical removal while minimizing the risks of permanent postoperative deficits. This results in an improvement of survival and quality of life. PMID- 11487190 TI - Stroke in a young man with fibromuscular dysplasia of the cranial vessels with anticardiolipin antibodies: a case report. AB - A case of fibromuscular dysplasia (FMD), presenting with a non-hemorrhagic infarct is reported. Positivity of anticardiolipin antibodies suggested an immune response. A 40-year-old man presented with sudden onset of stroke, preceded by similar ischemic attacks. Computed tomography (CT) of the brain showed a recent non-hemorrhagic infarct in the left middle cerebral artery (MCA) territory and an old right MCA territory infarct. Serum was positive for anticardiolipin antibodies. These above findings were confirmed at autopsy. A portion of the internal carotid artery and the middle cerebral arteries on both sides revealed features of FMD, with thrombosis. This case suggests an immune mechanism for FMD, hitherto unobserved in the cerebral circulation. PMID- 11487191 TI - Clinical and genetic study of essential tremor in the Italian population. AB - Essential tremor (ET) is one of the most common movement disorders. The pathogenesis is as yet unknown, although a genetic cause has long been recognised. Clinical and molecular evidence suggested that the ET gene contains a CAG expanded region. We examined a cohort of 240 Italian ET patients, classified as familial (193 cases) and sporadic (47 cases). The clinical manifestations of ET patients confirmed that the disorder is characterised by a large phenotypic variability. Repeat expansion detection (RED) approach did not demonstrate large CAG expansions. Six families were genotyped with 12 microsatellites markers of 2p and 3q regions and analysed according to parametrical methods. Lod scores values obtained in these families excluded the association of ET with 2p and 3q loci. Our findings confirm the presence of genetic heterogeneity and suggest that at least a third locus is involved in the pathogenesis of familial essential tremor. PMID- 11487192 TI - Role of dopamine D1 receptors in the control of striatal acetylcholine release by endogenous dopamine. AB - In order to determine the role of dopamine (DA) D1 receptors in the control of striatal acetylcholine (ACh) transmission, we studied the effects of SCH 39166 (D1 receptor antagonist), alone or in combination with quinpirole (D2/D3 agonist) or PD 128,907 (D3 agonist) on ACh and DA release. Quinpirole reduced DA and ACh release; PD 128,907 decreased DA but not ACh release. SCH 39166 stimulated DA and decreased ACh release. Pretreatment with quinpirole reduced or prevented (depending on the dose) the stimulation of DA release while potentiating the decrease of ACh release elicited by SCH 39166. Similarly, SCH 39166 administered following PD 128,907 did not stimulate DA release, further decreasing ACh release. These results indicate that quinpirole or PD 128,907 affect the actions of SCH 39166 on DA and ACh release in opposite manner, counteracting the increase of DA release and potentiating the reduction of ACh release. These data support the tenet that endogenous DA exerts a stimulatory input on striatal ACh neurotransmission mediated by D1 receptors. PMID- 11487193 TI - The role of language areas in motor control dysfunction in Parkinson's disease. AB - We evaluated the differences in motor control organization between parkinsonian patients with (seven cases) and without (ten cases) gait disorder. We used positron emission tomography (O15-H2O-PET) to measure regional cerebral blood flow as a correlate for local neuronal activation. This has been assessed during repetitive joystick movements of the right hand, externally triggered using a metronome as an auditory cue. In patients with Parkinson's disease (PD) without gait disorder, the contralateral supplementary motor cortex and the bilateral cerebellum were activated, while in PD patients with gait disorder the contralateral Broca's area, the contralateral sensory motor cortex and the homolateral cerebellum were activated. Our results suggest that PD patients with gait disorder creates an internal verbal cue in order to control the output of the movement of joystick, supplying the loss of control of the supplementary motor cortex that is activated in patients without gait disorder. PMID- 11487194 TI - Perfusion ECD/SPECT in the characterization of cognitive deficits in Parkinson's disease. AB - Cognitive abnormalities have been reported in a large percentage of patients with Parkinson's disease (PD). Often cognitive changes are sub-clinical and involve frontal lobe function. In other occasions they develop into full dementia. Functional neuroimaging may help characterize these abnormalities. We have studied brain perfusion with SPECT and the tracer ECD in 44 PD patients, 22 presenting with normal cognitive function and 22 with clinical and neuropsychological signs of dementia. Compared with 21 healthy controls, demented PD patients showed significant perfusion decrements in all cortical areas, particularly temporal and parietal regions; in the non-demented cohort reductions were limited to the frontal lobe area. These results suggest that brain perfusion abnormalities are present in PD patients. It is speculated that different pathological mechanisms underlie perfusion differences. PMID- 11487195 TI - The status of dopamine nerve terminals in Parkinson's disease and essential tremor: a PET study with the tracer [11-C]FE-CIT. AB - Neuroimaging studies of the striatal dopamine transporter (DAT) are useful in the assessment of the dopaminergic system in Parkinson's disease (PD). We used positron emisson tomography (PET) and the tracer [11C]FE-CIT to measure DAT binding in the caudate nucleus and putamen of 31 patients with PD, 5 with essential tremor and 8 healthy control subjects. Of the patients with PD, 17 were drug naive, while the others were either on levodopa or dopamine agonist monotherapy. DAT binding was significantly reduced in the caudate nucleus and to a greater extent in the putamen of PD patients compared to both healthy controls and essential tremor individuals. No overlap was observed between putamen values in PD and normals. No differences were found between controls and essential tremor subjects. These data confirm that measurements of DAT binding can provide an accurate and highly sensitive measure of degeneration in the dopamine system in PD. PMID- 11487196 TI - Blockade of subthalamic glutamatergic activity corrects changes in neuronal metabolism and motor behavior in rats with nigrostriatal lesions. AB - We infused--for four weeks--a selective antagonist of the NMDA receptor, MK-801, into the subthalamic nucleus of rats bearing an evolving nigrostriatal lesion. The aim was to block the subthalamic overactivity resulting from the dopaminergic striatal denervation. The nigrostriatal lesion caused metabolic activation- increased activity of the mitochondrial enzyme succinate dehydrogenase--of basal ganglia nuclei, ipsilaterally to the lesion, along with contralateral rotational behavior. These phenomena were effectively counteracted by the blockade of glutamatergic transmission at the subthalamic level. Pharmacological manipulation of the STN, through selective drugs capable of modulating glutamatergic transmission, may therefore represent a valuable tool for the treatment of PD. PMID- 11487197 TI - The parkin gene and its phenotype. Italian PD Genetics Study Group, French PD Genetics Study Group and the European Consortium on Genetic Susceptibility in Parkinson's Disease. AB - Mutations of the parkin gene on chromosome 6 cause autosomal recessive, early onset parkinsonism. This is the most frequent form of monogenic parkinsonism so far identified. The associated phenotypical spectrum encompasses early onset, levodopa-responsive parkinsonism (average onset in the early 30s in Europe), and it overlaps with dopa-responsive dystonia in cases with the earliest onset, and with clinically typical Parkinson's disease in cases with later onset. Despite clinical features, Lewy bodies are not found at autopsy in brains of patients with parkin mutations. The parkin protein possesses ubiquitin ligase activity, which is abolished by the pathogenic mutations. PMID- 11487198 TI - Chronic delusional hallucinatory psychosis in early-onset Parkinson's disease: drug-induced complication or sign of an idiopathic psychiatric illness? AB - Chronic delusional psychosis with hallucinations (CDHP) is commonly assumed to complicate the later stages of Parkinson's disease, as a side effect of antiparkinsonian medication. We studied 7 patients with early onset PD, who had developed psychiatric manifestations consisting in CDHP after a few years of antiparkinsonian therapy. All patients underwent a neurological, psychiatric and brain imaging (CT or MRI) evaluation. Detailed clinical history was recorded in order to reveal prior psychiatric illness and to analyse the relationship between neurological disease, cognitive impairment and psychosis. Our findings suggest that CDHP occurring in patients with early onset PD, normal or slightly impaired cognitive functions and normal CT/MRI scans is invariably the expression of a coexisting psychiatric illness which prior to onset of the neurologic disease had not been correctly diagnosed and which has been disclosed by dopaminergic therapy. PMID- 11487199 TI - Presymptomatic tests in Huntington's disease and dominant ataxias. AB - Huntington's disease (HD) and dominant ataxias (SCA) represent neurodegenerative hereditary diseases dominantly transmitted for which a direct and accurate genetic test is now available for molecular confirmation and presymptomatic test. Predictive testing programs, according to published international guidelines, are available worldwide. A large number of subjects (n=165) required a predictive HD diagnosis, although only 36% completed the program flow-chart and received the final genetic result (26 had a positive, 34 negative result for mutation). In 4 cases, an allele of intermediate range (33-34 CAGs) was found. Two of these shared the intermediate allele with an expanded repeat. In this case, we estimated the patient's risk to have affected children over the usually reported 50%. In 4 cases, the presymptomatic diagnosis was requested by persons at-risk for SCA1 and SCA3/Machado-Joseph disease. There were no adverse events to results of both HD and SCA presymptomatic diagnoses. PMID- 11487200 TI - Chronic bilateral electrical stimulation of the subthalamic nucleus for the treatment of advanced Parkinson's disease. AB - Preliminary reports in patients with Parkinson's disease (PD) showed that subthalamic nucleus (STN) stimulation was able to reverse parkinsonian state. Since 1998 we evaluated the safety and the efficacy of STN stimulation in 7 patients affected by advanced PD. All patients were included using CAPIT protocol. Motor functions and quality of life were evaluated, before and after surgery, with UPDRS and PDQ38, respectively. At the 6-month follow-up, the off medication/on stimulation UPDRS motor score improved by 50.6% and the on medication/on stimulation by 20.3%. Motor fluctuations were reduced by 57.2% and dyskinesias by 73.5%. The total L-dopa equivalent daily dose was reduced by 40.7%. PDQ38 ameliorated by 49.9%. We did not observe any perioperatory complication and only mild and tolerable side effects after stimulation. PMID- 11487201 TI - Alterations in GAD67, dynorphin and enkephalin mRNA in striatal output neurons following priming in the 6-OHDA model of Parkinson's disease. AB - In the 6-hydroxydopamine (6-OHDA) rat model of Parkinson's disease, administration of a dopaminergic agonist sensitizes rats to a subsequent administration of dopaminergic drugs given days apart (priming). In situ hybridization was used to evaluate changes on striatal gene expression of rats primed three days previously with either L-dopa, SKF38393 or quinpirole. Double labeling was used to identify the neuronal population in which such alterations occurred. GAD67 and enkephalin mRNA were increased by the lesion whereas dynorphin mRNA was decreased as compared to the intact striatum. Priming with L dopa and SKF38393 significantly increased GAD67 mRNA in the lesioned striatum and reversed dynorphin mRNA reduction, as compared to drug-naive rats, whereas quinpirole failed to produce any effect. Enkephalin mRNA was not affected by priming. Results suggest that 6-OHDA lesion-induced adaptive changes on striatal gene expression are modified by priming. Priming brings striatal output neurons to a higher level of activity, which may explain the sensitized behavioral response observed following a dopaminergic agonist challenge. These changes are in relation to the different types of dopamine agonists utilized and suggest that modifications in gene expression induced by priming might be predictive of the dyskinetic potential of a drug. PMID- 11487202 TI - An abnormal striatal synaptic plasticity may account for the selective neuronal vulnerability in Huntington's disease. AB - A marked decrease in the activity of mitochondrial complex II (succinate dehydrogenase, SD) has been found in the brains of Huntington's disease (HD) patients. Here we have examined the possibility that SD inhibitors might produce their toxic action by increasing corticostriatal glutamatergic transmission. We report that SD inhibitors produce a durable augmentation of NMDA-mediated corticostriatal excitation (DANCE) in striatal spiny neurons, but not in striatal cholinergic interneurons. DANCE involves increased intracellular calcium, activation of MAP kinase ERK and is critically dependent upon endogenous dopamine (DA) acting via D2-like receptors. This pathological form of corticostriatal synaptic plasticity might play a key role in the regional and cell-type specific neuronal death observed in HD. PMID- 11487203 TI - Role of striatal acetylcholine on dopamine D1 receptor agonist-induced turning behavior in 6-hydroxydopamine lesioned rats: a microdialysis-behavioral study. AB - The effects of MK-801, a non-competitive N-methyl D-aspartate (NMDA) receptor antagonist, of quinpirole, a dopamine (DA) D2 receptor agonist, and of SCH 58261, an A2A adenosine antagonist, were studied on acetylcholine (ACh) release in the striatum of 6-hydroxydopamine (60HDA) lesioned rats and on turning behavior induced by the administration of the DA D1 agonist CY 208-243. Administration of CY 208-243 to 6OHDA lesioned rats induced turning behavior and dose-dependently stimulated ACh release. At the dose of 50 microg/kg, MK-801 failed to affect basal ACh, while at 100 microg/kg MK-801 reduced it; however, MK-801 (50 and 100 microg/kg) potentiated the turning behavior elicited by CY 208-243, but failed to affect the CY 208-243-induced increase of striatal ACh release. The administration of quinpirole induced low-intensity turning behavior and decreased basal ACh release; on the other hand, quinpirole potentiated the turning behavior induced by CY 208-243, but failed to affect the CY 208-243-elicited increase of ACh release. Finally, intravenous administration of SCH 58261 stimulated basal ACh release but not turning behavior; SCH 58261, however, potentiated turning behavior induced by CY 208-243, while failing to affect the D1-elicited increase of ACh release. These results indicate that potentiation of D1-dependent turning behavior by MK-801, quinpirole and SCH 58261 is not mediated by a reduced ability of D1-agonists to stimulate ACh release from the denervated striatum. PMID- 11487205 TI - Deep brain stimulation of the subthalamic nucleus: selection of patients and clinical results. AB - Deep brain stimulation of the subthalamic nucleus has been proved to be an effective treatment for advanced Parkinson's disease when therapeutical strategies have failed. A correct selection of candidates for surgery is fundamental to obtain a good clinical effect. In this study we present our protocol of patient selection. In addition we report the data relative to the different causes of exclusion and the clinical efficacy of the electrical stimulation of the subthalamic nucleus at 3 months and 1 year follow-up. PMID- 11487204 TI - Decreased platelet glutamate uptake and genetic risk factors in patients with Parkinson's disease. AB - Genetic risk factors seem to play a role in sporadic Parkinson's disease (PD), maybe triggering oxidative stress and excitotoxicity within substantia nigra. However, genetic factors act at systemic level: reduced activity of mitochondrial enzymes and decreased glutamate uptake have been shown in platelets from PD patients. In this study we investigated glutamate uptake in platelets from 38 sporadic PD patients, 13 patients with parkinsonian syndromes and 28 controls and assessed polymorphisms of alpha-synuclein and ApoE genes. A 48% reduction of glutamate uptake p)<0.0001) was observed in PD patients which, with respect to control groups, correlated with the disease severity (r = -0.44, p < 0.05). Genetic studies of this population did not show differences between PD and controls, nor correlations with platelet glutamate uptake. PMID- 11487206 TI - Proton magnetic resonance spectroscopy (1H-MRS) of motor cortex and basal ganglia in de novo Parkinson's disease patients. AB - Proton MR spectroscopy (1H-MRS) has been previously performed in Parkinson's disease (PD) and parkinsonian syndromes to evaluate in vivo concentrations of basal ganglia and cerebral cortex metabolites such as N-acetylaspartate (NAA), choline (Cho), and creatine (Cr). However, this technique has never been used to evaluate motor cortex in untreated PD patients. In this study, single-voxel 1H MRS of basal ganglia and motor cortex was carried out in 10 de novo patients with PD and 10 age-matched healthy controls. A significant reduction in the NAA/Cr ratio was observed in the motor cortex of PD patients compared with controls (p)<(0.01). Basal ganglia spectra did not allow any evaluation due to the presence of artefacts related to inorganic paramagnetic substances. The motor cortex reduction of the NAA/Cr ratio in de novo PD patients may reflect an altered neuronal functioning due to a loss of thalamocortical excitatory inputs and may represent an in vivo marker for the diagnosis of PD. PMID- 11487207 TI - Interaction between dopamine and adenosine A2A receptors as a basis for the treatment of Parkinson's disease. AB - The adenosine A2A receptor antagonist SCH 58261 increases the turning behaviour induced by L-dopa in unilaterally 6-hydroxydopamine (6-OHDA)-lesioned rats. In this study we have evaluated the effect of a chronic intermittent administration of L-dopa or SCH 58261 plus L-dopa on turning behaviour. Chronic intermittent administration of SCH 58261 plus L-dopa produced a stable turning behaviour during the course of the treatment, whereas L-dopa alone produced a progressive increase in turning behaviour. Moreover, repeated administration of SCH 58261 failed to produce tolerance to its ability to potentiate L-dopa-induced turning behaviour. The results indicate that SCH 58261 is effective after chronic administration and suggest that SCH 58261 plus L-dopa, differently from Ldopa alone, does not produce alterations in motor responses during the course of the treatment. PMID- 11487208 TI - The parkin gene is not a major susceptibility locus for typical late-onset Parkinson's disease. AB - We investigated the parkin gene in 118 patients with typical Parkinson's disease (PD), i.e. in patients who had an onset of PD after the age of 45 years. The study group included 95 subjects with sporadic PD and 23 subjects from 18 families with autosomal recessive PD. No pathogenetic mutations in the parkin gene were detected either in familial or in sporadic patients. Our findings indicate that the parkin gene is not involved in the pathogenesis of classic late onset PD. PMID- 11487209 TI - Amantadine for dyskinesia in patients affected by severe Parkinson's disease. AB - 20 patients (12 men and 8 women, mean age 65 years) affected by severe Parkinson's disease (PD) with peak-dose and/or diphasic dyskinesias or painful dyskinesia were treated with amantadine (300 mg/day) as adjunctive therapy to current levodopa, carbidopa and dopaminoagonist. UPDRS (Unified Parkinson's disease rating scale), dyskinesias rating scale (DRS) and IGA (investigator global assessment) scale were used to evaluate the severity of PD symptoms during follow-up. After 15 days with amantadine treatment all patients improved with an average 38% reduction in dyskinesias (p<0.001). After 2-8 months. amantadine was withdrawn in all patients. After amantadine withdrawal, 2 patients experienced severe hyperthermia (39 degrees C and 40 degrees C). No difference was found between end of treatment dyskinesia scores and final withdrawal scores (p<0.5). In the two patients with hyperthemia amantadine was reintroduced; after four days hyperthermia subsided and amantadine was finally tapered over 15 days without further adverse reactions. PMID- 11487210 TI - CSF phosphorylated tau is a possible marker for discriminating Alzheimer's disease from dementia with Lewy bodies. Phospho-Tau International Study Group. AB - Tau and beta-amyloid (1-42) (Abeta42) are two independent markers for the early diagnosis of Alzheimer's disease (AD). In the present study, biochemical markers were validated as tools for differential diagnosis between AD and dementia with Lewy bodies (DLB). Tau, Abeta42 and phospho-tau (181P) were measured in cerebrospinal fluid (CSF) from controls (n=40) and from patients with AD (n=80) or DLB (n=43) using the HT7-AT270 assay (prototype version). In comparison with AD, in DLB no differences were found for Abeta42 and lower phospho-tau. ROC analysis was used to compare the discriminatory power of total tau with that of phospho-tau. The area under the curve (AUC) amounted to 0.782 +/- 0.048 (mean +/- SE) for tau and to 0.839 +/- 0.042 for phospho-tau (p = 0.039) for differentiation of AD from DLB. The present results indicate that CSF phospho-tau may be a good marker for differentiation between AD and DLB. PMID- 11487211 TI - Stimulation of growth hormone release in multiple system atrophy, Parkinson's disease and idiopathic cerebellar ataxia. AB - Clonidine has been proposed to differentiate multiple system atrophy (MSA) from idiopathic Parkinson's disease (IPD), as it does not increase growth hormone (GH) release in MSA. We studied GH release in response to clonidine in 7 IPD patients, 6 MSA patients, 4 patients affected by idiopathic late-onset cerebellar ataxia (ILOCA) and 8 healthy controls. In addition, we investigated the effects of GH releasing hormone plus arginine (GHRH-Arg) on GH release in the same patients. Both clonidine and GHRH-Arg raised serum GH levels in all groups examined. Clonidine failed to differentiate MSA from IPD and ILOCA. GHRH-Arg showed a lower increase of serum GH in MSA patients than in other groups, even if such difference was not statistically significant. We suggest that stimulation of GH release with GHRH-Arg rather than clonidine could differentiate MSA from IPD and ILOCA, but this hypothesis would need to be confirmed by further investigations. PMID- 11487212 TI - Deep brain stimulation of subthalamic nucleus: behavioural modifications and familiar relations. AB - The present study investigated behavioural modifications and familiar relations in a group of 15 parkinsonian patients treated with bilateral deep brain stimulation of the subthalamic nucleus. In 70% of the patients, during the first months after surgery we observed a euphoric mood owing to motor signs amelioration, but a series of problems (fear to come back to the pre-operative condition, sense of failure, slowness in changing the old habits) arose when it was necessary to adjust the parameters of stimulation and the pharmacological therapy to obtain a stable clinical picture. The caregivers showed an aggressive behaviour as reaction to the persistent psychological dependence of the patients. This distressed condition could be the cause of the onset of incomprehensions within the couple. PMID- 11487213 TI - Cytogenetic analysis oxidative damage in lymphocytes of Parkinson's disease patients. AB - Several lines of evidence support the presence of DNA damage in somatic cells of Parkinson's disease (PD) patients due to the formation of free radical species. In order to detect spontaneous chromosome and primary or oxidative DNA damage, we performed the human lymphocyte micronucleus assay (HLMNA) and comet assay in 19 PD patients and 16 healthy controls. Compared with controls, PD patients showed a significant increase in: (I) spontaneous micronucleus (MN) frequency (p<0.001); (2) single strand break (SSB) levels (p<0.001); and (3) oxidized purine base levels (p<0.05). The chromosome damage and the increased levels of oxidized purine bases observed in our patients support the hypothesis of oxidative stress as a relevant factor in the pathogenesis of PD. PMID- 11487214 TI - Non-motor effects of deep brain stimulation of the subthalamic nucleus in Parkinson's disease: preliminary physiological results. AB - Although deep brain stimulation (DBS) is a clinically effective therapy for patients with advanced Parkinson's disease (PD), its physiological effects on the brain and possible actions on non-motor functional systems remain largely unknown. This study evaluated the effects of DBS of the subthalamic nucleus (STN) on neurophysiological variables and on cardiovascular physiology. Nine patients affected by PD undergoing chronic DBS of the STN have been studied. We performed electroencephalography (EEG), somatosensory (SEPs) and visual evoked potentials (VEPs), exteroceptive masseteric silent period and sympathetic skin response (SSR) studies with DBS ON and OFF. To assess the effects of stimulation on the cardiovascular system the tilt test and plasma renin activity were studied. When we turned the DBS OFF, both SEP N20 and the VEP P100 component increased significantly in amplitude whereas the SSR decreased in amplitude and increased in latency. Although plasma renin activity tended to increase with DBS OFF, its modification induced by postural changes and blood pressure values did not significantly differ with DBS ON and OFF. We conclude that DBS of the STN in PD, besides inducing a clinical improvement, induces several non-motor effects. PMID- 11487215 TI - Which target for DBS in Parkinson's disease? Subthalamic nucleus versus globus pallidus internus. AB - We selected 14 patients with advanced idiopathic Parkinson's disease (PD) and examined the clinical effects of STN DBS versus GPi DBS. Nine patients underwent bilateral STN DBS and five underwent bilateral GPi patients. All patients were followed for at least 12 months. The evaluation was performed on and off drug before surgery; on-drug/on-DBS and off-drug/on-DBS at 1, 3, 6 and 12 months after stereotactic surgery. At 1 and 3 months after surgery in off-drug/on-DBS condition, both groups showed an improvement in motor score (UPDRS III). Nevertheless, the results changed after long-term stimulation in the two groups. Chronic STN DBS is superior to GPi DBS in the amelioration of the clinical features and in the decrease of time spent in the off state. The efficacy in reduction of LID was comparable at 1 and 3 months after surgery, but the results were better in STN DBS after chronic stimulation. The L-dopa dose was reduced only in the STN group. PMID- 11487216 TI - Reduced L-dopa absorption and increased clinical fluctuations in Helicobacter pylori-infected Parkinson's disease patients. AB - We report that the area under the curve of L-dopa plasma concentration, following the administration of a single 250 mg L-dopa dose, is augmented after Helicobacter pylori (HP) eradication in six Parkinson's disease (PD) patients showing high IgG antibody titer against HP. A prolongation of L-dopa clinical benefit was also observed. We suggest that HP infection-activated gastric alterations may be responsible, at least in part, for the reported erratic efficacy of oral L-dopa therapy in some advanced PD patients. Given the high percentage of HP-positivity in the age cohorts including the largest prevalence of PD patients, we propose that HP eradication be recommended in all PD patients under L-dopa therapy. PMID- 11487217 TI - Subcutaneous continuous apomorphine infusion in fluctuating patients with Parkinson's disease: long-term results. AB - Fluctuations in motor disability and dyskinesias are the major problem in the long-term treatment of Parkinson's disease (PD). Many authors and ourselves have shown that by giving patients a continuous infusion of levodopa it is possible to control motor fluctuations. Levodopa can be administered continuously only by intravenous, intragastric or intrajejunal delivery. Continuous dopaminergic stimulation can be achieved more easily by infusing dopamine agonists subcutaneously. Apomorphine is a potent water-soluble dopamine receptor agonist that has been shown to successfully control motor fluctuation when subcutaneously infused in complicated parkinsonian patients. We report the clinical data of 30 PD patients having at least five years of treatment with subcutaneous continuous apomorphine infusion. PMID- 11487218 TI - Identification of a novel primary torsion dystonia locus (DYT13) on chromosome 1p36 in an Italian family with cranial-cervical or upper limb onset. AB - Primary torsion dystonia (PTD) is a clinically and genetically heterogeneous group of movement disorders, usually inherited in an autosomal dominant fashion. Three PTD loci (DYT1, DYT6 and DYT7) have been identified to date. However, in several PTD families linkage to the known loci has been excluded. We identified an Italian PTD family with 11 definitely affected members. Phenotype was characterised by juvenile or early-adult onset, prominent cranial-cervical and upper limb involvement, mild course and occasional generalisation. A genome-wide search performed in the family identified a novel PTD locus (DYT13) within a 22 cM interval on the short arm of chromosome 1, with a maximum lod score of 3.44 (theta = 0) between the disease and marker D1S2667. PMID- 11487219 TI - Epidemiology of multiple system atrophy. ESGAP Consortium. European Study Group on Atypical Parkinsonisms. AB - Multiple system atrophy (MSA) is a form of atypical parkinsonism with unknown etiology. The epidemiological studies conducted up to now on this disease are scarce. The incidence rate is about 0.6 cases per 100,000 persons per year. The prevalence rates show 4-5 cases per 100,000 persons. In Italy, about 4,900 prevalent cases have been estimated. The mean onset age is about 54 years; the median survival is 7-9 years. Only one case-control study has been performed on this disease. This study showed an increased risk of MSA associated with occupational exposure to organic solvents, plastic monomers and additives, pesticides and metals. Smoking habits seem to be less frequent in MSA cases (as in Parkinson's disease cases) than in healthy controls. Quinn's clinical criteria and those of the Consensus Conference promoted by the American Academy of Neurology are in fair agreement. We have performed a case-control study on 73 MSA cases, 146 hospital controls and 73 population controls. PMID- 11487220 TI - Differences in rodent cage changout results. PMID- 11487221 TI - On first laboratory animal medicine training program. PMID- 11487222 TI - Verification of bacterial killing effects of cage wash time and temperature combinations using standard penicylinder methods. AB - We conducted the present study to evaluate various time and hot-water temperature combinations necessary to kill three common bacterial species in standardized cultures on stainless steel penicylinders, in accordance with methods approved by the Association of Official Analytical Chemists. Exposure for no more than 2 sec to water at 82.2 degrees C killed all three bacterial species, as did exposure for 3 sec to water at 80 degrees C, 4 sec at 77.8 degrees C, and 5 sec at 75.6 degrees C. We conclude that temperatures in the range of 75.6 degrees C to 82.2 degrees C will effectively kill vegetative bacteria in a matter of seconds and that failure to kill these bacteria in cagewash operations, with belt travel times in minutes rather than seconds, is due to other factors that prevent the effective application of sufficiently hot water to the bacterial load. PMID- 11487223 TI - The influence of husbandry schedules on the number of embryos collected from superovulated mice. AB - We undertook this study to assess the effect of urine and fecal contamination in the cages of male mice on embryo production in females placed in these cages for breeding. Each superovulated female was placed with a stud male in a cage that was cleaned at 1, 24, or 168 h prior to breeding. Approximately 41 h after mating, embryos were flushed from the oviducts, and we determined the number of embryos and their developmental stages for each treatment group. The number of two-cell embryos collected did not differ significantly when cages were cleaned 1 or 24 h prior to mating the mice. However, significantly (p < 0.05) fewer embryos were recovered when cages were changed 168 h before the animals were mated. PMID- 11487224 TI - Eradication of simian retrovirus type D from a colony of cynomolgus, rhesus, and stump-tailed macaques by using serial testing and removal. AB - The markedly compromised health of animals in a macaque colony and the problematic interpretation of data from two drug safety assessment studies prompted a review of the effect of simian retrovirus type D on the drug development process at a Midwest pharmaceutical company. After reviewing relevant literature and consulting with an expert in simian retroviruses, we initiated a program of eradication. During a 16-month period, all cynomolgus (Macaca fascicularis), rhesus (Macaca mulatta), and stump-tailed (Macaca arctoides) macaques housed in the facility were evaluated as many as eight times for the presence of simian retrovirus type D by using serology, virus isolation, and/or polymerase chain reaction tests. All animals with positive test results were removed from the colony immediately. No test results indicative of simian retrovirus type D infection have occurred during the subsequent 2.5 years. We attribute the successful eradication and prevention of re-introduction of the virus to regular testing, purchasing animals from sources free of simian retrovirus type D, and assiduous application of procedures designed to prevent transmission between animals. PMID- 11487225 TI - A technique for restraining rodents during hindlimb interventions. AB - Recently interest has grown in the skeletal effects of external electrophysical modalities. To introduce these modalities into the rodent hindlimb, restraining the animals is often necessary. This report describes a novel restraint device that was highly successful when used on 10 Sprague-Dawley rats for 20 min daily for 6 days each week over a period of 12 weeks. The restrained animals gained less weight than did 10 cage control animals, suggesting that animals experienced stress. However, this weight loss was significant (p < 0.05) only during the first six weeks, suggesting a degree of acclimatization. Restraint had no observable effect on measures of bone mineral content of the hindlimb. The restraint device we developed restrained animals without the need for anesthetics, enabling our device to be used in future studies investigating the skeletal effect of external electrophysical modalities on the rodent hindlimb. PMID- 11487226 TI - Effect of laboratory acclimation on food and water consumption of pregnant sheep after fetal catheterization. AB - The objectives of this study were to determine 1) the time required for food and water consumption of late-gestation pregnant sheep to stabilize after a 6- to 7-h shipment by truck and 2) whether the duration of laboratory acclimation altered food and/or water consumption of pregnant sheep after fetal and maternal vascular catheterization. We used a semi-quantitative scale and a retrospective study design to determine food and water consumption as a function of acclimation time in post-shipping and post-surgery animals. These animals had been in our research facility for 2, 3, 4, and 5 or more days prior to surgical catheterization of the fetus and mother. We used a quantitative scale and a prospective study design to determine food and water consumption in post-surgery animals that had been in the laboratory for either 2 or > or = 7 days at the time of surgery. We used two- and three-factor repeated measures analyses of variance to determine the statistical significance of any differences. Although food and water consumption in post shipping animals were significantly (p < 0.001) lower on day 1 than other days, we attributed this difference to the fact that "day 1" was shorter than 24 hours because the animals arrived in the laboratory at noon. Further, the post-surgery decrease and subsequent recovery in food and water consumption did not depend on the duration of the acclimation prior to surgery. We conclude that differences in pre-surgery, post-transportation acclimation periods ranging from 48 hours to > or = 7 days do not affect post-operative recovery from fetal surgery in sheep. PMID- 11487227 TI - Zinc responsive dermatopathy in goats: two field cases. AB - Two adult female goats presented with symmetrical alopecia, rough hair coats, weight loss, and generalized unthriftiness. In light of the history and clinical signs, we suspected a mineral deficiency or endocrine imbalance. These animals were being fed a diet of alfalfa, a legume high in calcium, which can block zinc uptake. Serum zinc levels were low, and skin biopsies supported the diagnosis of zinc deficiency. Therefore, these findings suggest that the main factor blocking zinc absorption was increased dietary calcium. PMID- 11487228 TI - Caerulein-induced pancreatitis in Wistar rats. AB - We describe a refined model of intravenous caerulein-induced pancreatitis by using osmotic infusion pumps in the conscious unrestrained Wistar rat. The volume of caerulein required for the 6-h infusion is loaded into PE-55 catheter tubing attached to an Alzet (Alza Corporation, Palo Alto, CA) implantable osmotic pump that has been primed with saline. The technique has reliably induced mild edematous pancreatitis, which we verified histologically. Our refined model has the advantages of unrestrained animals, reduced animal handling and acclimation, and decreased cost. PMID- 11487229 TI - Metastatic large intestinal adenocarcinoma in two rhesus macaques (Macaca mulatta). AB - Spontaneous colonic adenocarcinomas were diagnosed in two rhesus macaques (Macaca mulatta). Although both tumors caused partial obstruction of the colon, they were histologically different. One resembled the commonly described "napkin-ring" adenocarcinoma similar to those found in the descending and sigmoid colon of humans. The other neoplasm, a "tubular" lesion, invaded the full thickness of the colonic wall at the ileocecal-colonic junction. Both tumors were associated with metastasis, which is uncommon in the rhesus monkey. PMID- 11487230 TI - Endocarditis and meningitis caused by Streptococcus suis after cardiac surgery in a sheep. AB - A female Dorset-cross sheep developed labored respirations and was anorexic, weak, ataxic and febrile (42.0 degrees C) 15 days after implantation of a cardiac device. Clinical pathologic evaluation identified a mild leukocytosis (11,800 cells/microl), neutrophilia (8,969 cells/microl), and hypokalemia (2.9 mmol/L). Despite intensive therapy, the animal was euthanized 7 days later. Necropsy of the sheep revealed meningitis and vegetative endocarditis associated with the cardiac device. Blood cultures collected antemortem and tissue cultures collected at necropsy yielded Streptococcus suis. Histopathology confirmed the diagnosis of bacterial valvular endocarditis and meningoencephalitis. A variety of environmental and host factors are proposed as contributing to the infection, and the zoonotic potential of S. suis is discussed. PMID- 11487231 TI - Quantitative competitive PCR. PMID- 11487232 TI - The magnitude and duration of the analgesic effect of morphine, butorphanol, and buprenorphine in rats and mice. AB - This study was designed to determine the magnitude and duration of the analgesic effect of three commonly used opioids: buprenorphine (0.5 mg/kg for rats; 2.0 mg/kg for mice), butorphanol (2.0 mg/kg for rats; 5.0 mg/kg for mice), and morphine (10 mg/kg for rats and mice). We used two standard tests, the hot plate and tail flick assays, to measure opioid analgesia in 62 male, 200 to 300 g Sprague-Dawley rats and 61 male, 25 to 35 g ICR mice. We obtained five baseline measurements then administered the drugs subcutaneously. Morphine gave the highest analgesic effect and was intermediate in duration (2 to 3 h in rats and mice) of analgesia. Butorphanol provided the lowest level of and shortest (1 to 2 h in rats and mice) analgesia. Buprenorphine had an intermediate analgesic effect and the longest duration (6 to 8 h in rats and 3 to 5 h in mice). In light of our results, we recommend the use of morphine (with frequent redosing) for severe pain, butorphanol for mild pain of short duration, and buprenorphine for mild to moderate pain of increased duration. The dosing intervals suggested by our study are 2 to 3 h for morphine in both rats and mice, 1 to 2 h for butorphanol in both rats and mice; and 6 to 8 h in rats and 3 to 5 h in mice for buprenorphine. PMID- 11487233 TI - Evaluation of an anesthetic regimen for retroorbital blood collection from mice. AB - Many Institutional Animal Care and Use Committees require anesthetization of mice for retroorbital blood collection procedures. Optimal anesthetic regimens should provide chemical restraint and adequate analgesia for a sufficient but otherwise minimal duration. This study was conducted to evaluate the effectiveness, safety, and practical utility of the general anesthetic combination of ketamine and medetomidine with and without the topical anesthetic proparacaine. In addition, we sought to evaluate the reversal of general anesthesia with atipamezole. The findings suggest that the combination of ketamine, medetomidine, proparacaine, and atipamezole is a safe and humane anesthetic regimen for retroorbital collection of blood samples from mice. PMID- 11487235 TI - Artificial turf foraging boards as environmental enrichment for pair-housed female squirrel monkeys. AB - We investigated the use of artificial turf foraging boards to determine if providing captive squirrel monkeys an opportunity for semi-natural foraging behavior would 1) alter the monkeys' time budget to better approximate that seen in wild populations, 2) reduce the stereotypic, self-injurious, and aggressive behavior occasionally seen in captive squirrel monkeys, and 3) provide sustained enrichment. Five groups of pair-housed female squirrel monkeys were videotaped the week prior to, the week following, and for 2 weeks during the enrichment phase, when treat-enhanced boards were provided for 2 h daily. During the first 30 min of daily enrichment, inactivity declined 35.3%, locomotion increased 3.8%, and board-related behaviors occupied 36.3% of the activity budget; these changes were not evident after 1.5 h. Stereotypic behavior (pacing, headswinging, tailchewing) and aggression were not altered by the foraging opportunity. The foraging board retained the interest of the subjects across 2 weeks in the same daily pattern. Use of the foraging board altered the squirrel monkeys' time budget to become more like activity patterns seen in wild populations. PMID- 11487234 TI - Toxicity evaluation of prophylactic treatments for mites and pinworms in mice. AB - The detection of external and internal parasites in laboratory mice is a particularly problematic aspect of animal health evaluation. Because these organisms must be detected by direct examination of the feces or hair coat, low level infestation or sporadic shedding can make them difficult to detect, thereby undermining confidence that negative reports are truly negative. Prophylactic treatment of suspect colonies with anthelminthics and/or insecticides may therefore be indicated under some circumstances. However, when considering the use of prophylactic treatments, the potential for toxicity is an important factor, especially in genetically modified strains of mice. To evaluate the potential toxicity of prophylactic anti- parasitic treatments on strains of mice that are commonly used as experimental models and in genetic engineering in our facility, we surveyed a number of strains and ages of mice for toxic reactions during treatment regimens that combine anthelminthic and anti-acaricidal agents. Three experimental protocols (ivermectin, piperazine, and dichlorvos in combination; ivermectin alone; and fenbendazole/permethrin or fenbendazole/dichlorvos) were evaluated. Our data suggest a potential for toxicity associated with these treatments and indicate to us that prophylactic treatment regimens should be initiated with caution. PMID- 11487236 TI - Use of the feed additive sodium hexametaphosphate to prevent dental calculus in squirrel monkeys (Saimiri spp.). AB - Dental calculus and associated periodontal disease are serious clinical problems in captive squirrel monkeys. Calculus begins to appear as early as 2 years of age, with subsequent development of periodontal disease, dental abscessation, tooth loss and other sequelae. When used as a feed additive, sodium hexametaphosphate (HMP) retards the growth of calculus on previously cleaned teeth in rhesus monkeys, lemurs, and other species. We wanted to determine whether HMP would reduce dental calculus in squirrel monkeys (Saimiri spp.) whose teeth had not been pre-cleaned. The study animals were divided into two groups. One received a standard diet; the other received an identical diet containing the HMP additive at a concentration effective in other primate and non-primate species that had received dental cleaning prior to treatment with HMP. Teeth were graded for extent of calculus formation at the start of the study and at 3 and 6 months during HMP treatment. We compared the results from the two groups both by total score per animal and according to tooth type (e.g., incisors versus incisors in test and control groups). At the end of 6 months, dental calculus did not differ significantly between the experimental groups. Therefore, we conclude that HMP is ineffective in squirrel monkeys with preexisting dental calculus. PMID- 11487237 TI - Condyloma acuminatum in a cynomolgus monkey (Macaca fascicularis). AB - Condyloma acuminatum was found on the penile shaft and glans of a wild-caught cynomolgus monkey. The characteristic cauliflower-like, digitated lesions corresponded histopathologically with epidermal hyperplasia, hyperkeratosis, hypergranulosis, pallor of the mid-spinous zone, and crowding of keratinocytes. Electron microscopy revealed 10 intranuclear viral particles. In situ hybridization studies using human DNA probes (pan-Human Papillomavirus [HPV] 6/11, HPV 16/18, and HPV 31/33) all failed to hybridize. Polymerase chain reaction using papillomavirus probes MY09, MY11, GP5, and GP6 failed to amplify HPV DNA. We conclude that this lesion likely is a viral-induced condyloma but that the virus is not detectable by using presently available human or monkey papillomavirus probes. PMID- 11487238 TI - Radiotelemetry catheter implantation: surgical technique and results in cats. AB - Arterial blood pressure in cats can be measured by direct or indirect techniques. Most direct measurement techniques require sedation or anesthesia, which decreases blood pressure. The restraint and human intervention needed for indirect techniques cause blood pressure to increase. These limitations make the accurate assessment of blood pressure and the response to antihypertensive medications difficult to interpret. Radiotelemetry allows for direct measurement of the arterial blood pressure and heart rate without sedation, anesthesia, or animal handling during measurement. A surgical technique for the placement of radiotelemetry catheters and the complications associated with the technique have not been described in cats. Our purpose is to provide a detailed description of the placement of the modified implant model TA11PA-C40 (Data Sciences International), with accompanying illustrations, and to discuss complications associated with the procedure. Radiotelemetry catheters were surgically placed in the left femoral artery of 12 cats. The surgical procedure was completed within 50 min, and no surgical complications occurred in any cat. The radiotelemetry catheters were used for an average of 6.5 weeks (range, 5.5 to 9.5 weeks). This technique allows for long-term monitoring of ambulatory blood pressure and heart rate in the research setting. This methodology is especially useful for studies of the pathophysiology of hypertension and assessment of the efficacy of antihypertensive medications. PMID- 11487239 TI - A laryngoscope designed for intubation of the rat. AB - We successfully intubated 257 rats with a laryngoscope that we designed for this purpose. Orotracheal intubation with this laryngoscope can be performed quickly and without harm to the animal. This instrument provides direct visualization of the vocal cords, allowing rapid, safe intubation of the rat. Maintenance of an adequate airway with endotracheal intubation is superior to tracheostomy for repeated experiments, and therefore this laryngoscope makes such studies easier and safer to perform. PMID- 11487240 TI - Isoflurane anesthesia for guinea pigs (Cavia porcella) in a stereotaxic surgical apparatus. AB - Isoflurane (1-chloro-2,2,2-trifluoroethyl difluoromethyl ether) is a potent general anesthetic agent that is highly adaptable to use in small laboratory animal species. Respiratory delivery is easily accomplished in these animals, even during restraint in a stereotaxic apparatus. The ease of administration, high predictability of action, and ability to finely control the depth of anesthesia make isoflurane anesthesia far superior to regimes involving injectable anesthetics. The wide margin of safety of isoflurane is a key indicator of the superiority of this product to all other anesthetic agents. The initial cost associated with the delivery system is a drawback, but the reliability and safety factors outweigh the hardware costs. PMID- 11487241 TI - Colitis in a female tamarin (Saguinus mystax). AB - An adult wild-caught female tamarin (Saguinus mystax) housed in a biomedical research facility was found moribund and extremely dehydrated, with severe diarrhea. She initially responded to supportive therapy but died 3 days later. Necropsy findings included hyperemia of the colonic mucosa, mesenteric lymphadenopathy, acanthocephalid parasites (Prosthenorchis elegans) embedded in the mucosa of the terminal ileum and cecum, and free filarid nematodes (Dipetalonema sp.) in the abdominal cavity. Campylobacter sp. was recovered from the colon. With the exception of changes associated with the parasites, significant histologic changes were limited to the colon. Changes consistent with acute enteric viral infection were found against a background of chronic inflammation. Enterocytes were variably hypertrophied, degenerate, and attenuated. Numerous epithelial syncytial cells were present, and some affected cells (uni- as well as multinucleated) contained intranuclear inclusions. Lymphoid follicles were mildly depleted and contained rare syncytia. Measles (Morbillivirus) infection was confirmed by using immunohistochemistry, reverse transcriptase polymerase chain reaction, and Southern blot analysis. In contrast to the cutaneous rash, respiratory involvement, and low mortality characteristic of Old World monkeys with measles, severe diarrhea with high mortality occurs in New World monkeys with this disease. In addition, our case differs from previous reports of measles in New World monkeys in that syncytial cell formation apparently was limited to the colon of our animal. PMID- 11487242 TI - Southern and Northern hybridization. PMID- 11487243 TI - Macroenvironmental relative humidity and bedding in isolator cages. PMID- 11487245 TI - Pain and distress: Where is the role for clinical judgement? PMID- 11487246 TI - Food and water restriction protocols: physiological and behavioral considerations. AB - Food and water restriction protocols are common in animal research, yet they often elicit discussion and controversy among institutional animal care and use committee members who review them. Determining a single standard by which all restriction protocols can be evaluated or performed may not be realistic. However, information about the physiologic and behavioral impact of food and water restriction can provide a basis for making rational judgments about these issues in general. This review will discuss the physiologic and behavioral consequences of food or water deprivation periods of 24 h or less and of chronic restriction schedules, with special reference to protocols that use food or water restriction as a motivational tool for behavioral training. PMID- 11487247 TI - An evaluation of analgesic regimens for abdominal surgery in mice. AB - This study was designed to evaluate the efficacy of several analgesic regimens for use after intraperitoneal implantation of telemetry transmitters in mice. The lengths of time required for postoperative recovery of food and water intake, locomotor activity, and core temperature of mice that did not receive postsurgical analgesic medication were compared to those of mice that were given either an analgesic in the drinking water or buprenorphine injections. Many measured variables were not substantially altered by analgesic medications. However, ibuprofen-treated mice demonstrated significantly greater locomotor activity on days 2 through 5 after surgery and a more rapid return to stable postsurgical levels of activity and water intake as compared to those in untreated mice. These changes are consistent with potential analgesic efficacy of the ibuprofen treatment regimen. Buprenorphine injections elicited hyperactivity, hyperthermia, and reduced food and water intake during both the immediate postsurgical recovery period and after apparent recuperation from surgery, as compared to effects observed in saline-treated mice. Evaluating the effect of analgesic regimens on postsurgical changes in physiologic and behavioral variables can be useful in assessing the efficacy of analgesic treatments, but some changes may indicate pharmacologic effects that do not reflect pain relief. PMID- 11487248 TI - A method of video-assisted thoracoscopic surgery for collection of thymic biopsies in rhesus monkeys (Macaca mulatta). AB - To support an infectious disease study, we developed a surgical procedure to collect serial thymic biopsies in rhesus monkeys (Macaca mulatta). In many instances in which thymic tissue is required from living animals, open surgical approaches (thoracotomies) are used, which result in greater postoperative pain and longer recovery periods than those associated with thoracoscopic procedures. Our intent was to develop a surgical procedure that allowed serial biopsy of the thymus with minimal surgical morbidity. We modified a previously published experimental method of thoracoscopic total thymectomy in the dog to collect thymic biopsies in M. mulatta. Of the 15 animals evaluated, 8 underwent two biopsy procedures separated by a 5- to 6-week interoperative interval. The other seven animals underwent a single biopsy procedure. Thymic tissue was collected successfully during all procedures, with an average surgical time of 15 min. No significant intra- or postoperative complications were noted, and the animals recoveries from the surgical procedures were uneventful. Minimally invasive thoracoscopic surgical techniques can be used successfully to collect thymic tissue from adult and juvenile rhesus monkeys with minimal surgical morbidity. PMID- 11487249 TI - The thin blue line: a review and discussion of aseptic technique and postprocedural infections in rodents. AB - A basic tenet of animal welfare philosophy is that pain and distress must be minimized whenever possible without interfering with the goals of the research. Aseptic technique during surgical procedures is essential to prevent pain and distress associated with post-procedural infections. However, many investigators have found that applying the aseptic techniques used for large animal and human surgery is not always practical when performing surgery on small rodents. Furthermore, the efficacy of some of these techniques for preventing post procedural infections has been questioned. This review examines what is known about the development of postprocedural infections in animals and humans and the methods used to prevent them. Detection of postprocedural infections in rodents can be difficult unless objective measurements of physiologic indices are made. These measurements should be used experimentally to assess the relative benefits of various methods for preventing postprocedural infections. Measures of contamination, such as quantitative bacterial cultures, also can be used; however, they do not reliably predict infection rates. Much of the dogma about decontamination of skin and hair prior to surgery is not supported by valid experimental evidence. Hair removal may not be necessary. Alcohol may in fact be a better disinfectant than is often credited. Draping should be used when it contributes to the maintenance of the sterile field, but when it does not, modification of surgical technique may provide more protection from infection than the drape does. The contribution of surgical technique to the prevention of postprocedural infections is probably equal to that of aseptic technique. Further research needs to be done to assess various aseptic techniques for use in rodent surgery. PMID- 11487250 TI - Evaluation of epidural morphine for postoperative analgesia in ferrets (Mustela putorius furo). AB - We evaluated the analgesic efficacy of epidural morphine for relieving postoperative pain in domestic ferrets by evaluating behavior and fecal cortisol concentrations. The 12 laboratory-reared, intact, female, domestic ferrets were anesthetized then underwent ovariohysterectomy and bilateral anal sacculectomy. Using a double-blind procedure, we provided epidural morphine (0.1 mg/kg) to six ferrets and epidural saline (0.1 mL/ferret) to the remaining animals prior to surgery. Compared to the animals that received saline, the morphine-treated ferrets were more likely to have attenuated pain responses, and they returned more rapidly to preoperative behavior. Although fecal cortisol concentrations during the first 24 h after surgery increased in all animals, the increase was statistically significant only in the ferrets that received saline epidurals. These data suggest that morphine epidurals administered to ferrets prior to surgery may attenuate both the physiologic and behavioral manifestations of surgically induced pain. PMID- 11487251 TI - Isoflurane anesthesia in the African clawed frog (Xenopus laevis). AB - Isoflurane is one of the safest and most accepted anesthetic agents for reptiles, birds, and mammals. It has also been used in terrestrial amphibians. The use of inhalation agents in an entirely aquatic frog presents a new dilemma for delivery in contrast to terrestrial species. The African Clawed Frog respires by using both transcutaneous gas exchange and air breathing. These frogs remain submerged for long periods of time, thus making standard inhalation techniques impractical. We tested five methods for delivering isoflurane: 1) bubbling isoflurane and oxygen in the water, 2) intracoelomic injection, 3) subcutaneous injection, 4) intramuscular injection, and 5) topical application. For the topical application, we developed a simple technique by using an absorptive pad with a vapor-barrier backing, saturating the pad with the liquid isoflurane, and placing the pad on the back of the frog while it was confined in a plastic bowl. Although two of the three injectable routes induced anesthesia, only the topical route produced rapid induction with consistent, safe recovery. Bubbling isoflurane with oxygen into water was unsuccessful. Topical application of isoflurane was most successful and appears to be a safe and practical method that can be used as an alternative to tricaine methylsulphonate, hypothermia, or other methods for anesthetizing African Clawed Frogs. PMID- 11487252 TI - The effect of propofol administered intravenously on appetite stimulation in dogs. AB - Anorexia is defined as diminished appetite or aversion to food. Clinical manifestations of anorexia have multiple etiologies, which include systemic illness, pain, fever, stress, metabolic disorders, and decreased palatability and learned aversion to food. Disorders of appetite are common in companion and laboratory animal medicine. Anecdotal evidence and personal experience suggest that propofol (2, 6-diisopropylphenol), when given intravenously at subhypnotic doses, causes acute appetite stimulation in dogs. The establishment of a dose response effect could have important clinical applications; therefore, this study attempts to qualify and quantify the effect of propofol on appetite stimulation in healthy young adult dogs. Six purpose-bred male dogs (age, 6 months) were obtained from a Class A vendor. Dogs were housed individually and provided water ad libitum throughout the study period. All dogs were fed ad libitum to ensure that test conditions and degree of satiety were identical. Each dog was assigned randomly to either an experimental group or control each day of the study. The experimental groups received single bolus intravenous injections of propofol at different dosage levels (0.5, 1.0, 1.5, 2.0, or 3.0 mg/kg of body weight), and the control group received saline. The administrator was blinded to the animals identification and dose. Dosages greater than 3.0 mg/kg resulted in profound sedation and ataxia, which physically inhibited the dogs from obtaining the food; therefore 3.0 mg/kg was the highest dose tested. Dogs were weighed daily to ensure accurate dosing. Dosing was performed at the same time each day to minimize variability. Food intake amounts were recorded at 15, 30, 60, 120, and 1440 min after injection. Food intake was expressed as [food intake (g)/ body weight (kg)/ unit time (min)]. After a 1-w rest period, the study was repeated. Data were analyzed with a type RBF-65 randomized-block factoral design (ANOVA). Each dog served as its own control. The two experiments were analyzed separately, and a P-value of less than 0.05 was used to declare statistical significance. A significant (P, 0.05) increase in food consumption was observed solely during the 0-to-15-min time interval; no significant increase in food consumption was observed at any other time point. This data supports propofols appetite stimulating effect in the initial 15 min after injection. Additional studies are required to explore the mechanism for this effect and to determine whether it occurs in other species. PMID- 11487253 TI - Use of ultrafiltration probes in sheep to collect interstitial fluid for measurement of calcium and magnesium. AB - Studies of calcium and magnesium changes in living animals usually involve blood, urine, and fecal samples. These samples provide only information on whole-body averages and give no indication of differences between tissues. Ultrafiltration probes were developed to sample interstitial fluid from muscle, bone, and subcutaneous tissue of sheep to provide a tool for investigating tissue differences in calcium and magnesium concentrations. The potential of the probes for mineral distribution studies was demonstrated in sheep by using infusion of a calcium gluconate solution. PMID- 11487254 TI - Improving the environment of mice by using synthetic gauze pads. AB - We conducted a study to evaluate the use of synthetic gauze pads for improving the environment of mice. To evaluate differences in clinical and pathology parameters, we used two treatment groups of mice, which were housed with or without gauze pads. The mice were assigned to the study at 5 to 7 weeks of age, and the study lasted 1 year. The mice were housed individually in stainless-steel ventilated cages with wire-mesh floors. Clinical observations, body weights, and food consumption were recorded frequently during the study. A complete necropsy, with histopathologic evaluation of tissues and collection of blood for clinical pathology, was performed at completion of the study. The mice with gauze pads preferred to rest on them. In addition, these mice showed a statistically significant reduction in food consumption, but their body weights and weight gains did not differ from those of animals without gauze pads. Synthetic gauze pads provide an improved environment for mice housed in cages with wire floors and may produce reduced food consumption. Gauze pads in the cages of mice do not seem to influence body weight gain, clinical signs, clinical pathology, or morphologic pathology. PMID- 11487255 TI - Postnatal growth in a mouse genetic model of classical phenylketonuria. AB - Because of the ethical problems of withholding dietary treatment from patients with phenylketonuria, effectively studying long-term effects such as postnatal growth rates is difficult or impossible. The only literature available on the growth rate of phenylketonurics dates from the era before effective dietary treatment regimens were instituted routinely. Although some of these studies suggest that these patients have a reduced growth rate, whether this is a consistent feature of the untreated phenylketonuric is unclear. The mutant mouse line BTBR-Pahenu, a genetic model for human phenylketonuria, provides an opportunity to conduct studies that will clarify this issue. In this study, the weights of newborn mice were monitored from about 7 to 40 days after birth. Comparison to heterozygous, sex-matched littermates revealed a reduction in the size of homozygous mutants throughout the study. Therefore, reduced postnatal growth is an abiding feature of phenylketonuria in this mouse model. This finding not only helps to document the association between size reduction and untreated phenylketonuria but also sets the stage for studies designed to investigate the means by which size reduction occurs. PMID- 11487256 TI - Cholesterol granulomas in mice deficient in apolipoprotein E. AB - Three 12- to 16-month-old female B6,129 apolipoprotein E-deficient mice were presented for necropsy because of bilateral masses in the gluteal region. Histopathologic examination revealed cholesterol granulomas extending from the superficial dermis to the underlying gluteal muscles. Microscopic granulomas were found on the face, perivaginal tissues, mesenteric lymph nodes, spleen, joints, kidneys, and choroid plexus. Other lesions included severe atherosclerosis of the aortic valves, aorta, pulmonary artery, and renal artery. Here we show that macroscopic cholesterol granulomas can develop in apolipoprotein E-deficient that receive a normal rodent diet. PMID- 11487257 TI - Clinical implications of expression of vascular endothelial growth factor in metastatic lesions of ovarian cancers. AB - Vascular endothelial growth factor (VEGF) has been identified as an important factor for tumour angiogenesis, which is essential for the growth, invasion and metastasis of solid tumours. Significantly increased VEGF level from the primary tumour to the metastatic lesion of ovarian cancers was found in 8 of 30 cases. The 24-month survival rate of the patients with significantly increased VEGF level was extremely poor (0/8 = 0%) in comparison with that of patients with no change in the level (15/22 = 68%) from the primary tumour to the metastatic lesion. This indicates that VEGF may contribute to the advancement of metastatic lesions, and that VEGF level in metastatic lesions may be a prognostic indicator. PMID- 11487258 TI - Pharmacokinetics of anastrozole and tamoxifen alone, and in combination, during adjuvant endocrine therapy for early breast cancer in postmenopausal women: a sub protocol of the 'Arimidex and tamoxifen alone or in combination' (ATAC) trial. AB - The ATAC trial evaluates in a randomized, double-blind design, Arimidextrade mark (anastrozole) alone or in combination with tamoxifen, relative to tamoxifen alone as 5-year adjuvant treatment in postmenopausal women with early breast cancer. Patients included in the pharmacokinetic (PK) sub-protocol had been in ATAC for > or =3 months, taking their medication in the morning and were 100% compliant for the preceding 14 days. Blood samples were collected 24 +/- 4 h after last dose. Trough (C(min)) plasma concentrations of anastrozole, tamoxifen and desmethyltamoxifen (DMT) were measured by validated methods. The PK results were based on a total of 347 patients (131 anastrozole (1 mg o.d.), 111 tamoxifen (20 mg o.d.), 105 anastrozole and tamoxifen (1 and 20 mg o.d. respectively)). The geometric mean steady-state trough plasma concentrations of tamoxifen and DMT were statistically equivalent in patients receiving tamoxifen alone or in combination with anastrozole: geometric mean tamoxifen = 94.8 ng ml(-1)and 95.3 ng ml(-1)in tamoxifen alone and combination groups, respectively; geometric mean DMT = 265.1 and 277.6 ng ml(-1)in the tamoxifen and anastrozole and tamoxifen groups, respectively. The geometric mean anastrozole levels were 27% lower (90% Cl 20-33%;P< 0.001) in the presence of tamoxifen than with anastrozole alone. Baseline plasma oestradiol levels were not obtained in the PK sub-protocol, however, such information was available from a similar ATAC sub-protocol, which evaluated bone mineral density. Mean oestradiol levels were 21.3, 19.3, and 21.6 pmol l(-1)prior to treatment and 3.7, 20.9 and 3.6 pmol l(-1)after 3 months in the anastrozole, tamoxifen, and combination groups, respectively (n = 167). On treatment values were below the detection limit (3 pmol l(-1)) in 43.6 and 38.5% of the anastrozole alone and anastrozole in combination with tamoxifen groups, respectively. As a result of (a) the lack of effect of anastrozole on tamoxifen and DMT levels and (b) the observed fall in blood anastrozole levels having no significant effect on oestradiol suppression by anastrozole, we conclude that the observed reduction in anastrozole levels by tamoxifen is unlikely to be of clinical significance when anastrozole and tamoxifen are administered together. PMID- 11487260 TI - Longstanding survival without cancer progression in a patient affected by endometrial carcinoma treated primarily with leuprolide. AB - We report here a case of a patient affected by endometrial cancer and treated primarily with leuprolide, the surgical approach being unfeasible due to her compromised conditions. The therapy was continued for more than 6 years, and no progression of the disease was observed. During this period, some histological and immunohistochemical evaluations of the tumour (morphology, grading, proliferation and apoptotic index, E-cadherin expression) were performed. Furthermore, the expression of m-RNA for luteinizing-hormone releasing hormone (LHRH) receptors was determined. The results showed a discrepancy between some biological parameters of the tumour and its clinical characteristics. In fact, despite features suggestive of a progression of the cancer (such as the increase of both tumour grading and proliferating capacity (MIB-1), and a fall in the reparative process (appearance of mutated p53, reduced expression of both bcl-2 and c-erb-2) being detected, neither local invasion nor metastatic lesions were clinically observed. This discrepancy might be due to the maintenance of high levels of E-cadhezin. Moreover, since this tumour was shown to express mRNA for LHRH receptors, new evidence is provided about the favourable impact of LHRH analogue treatment in patients affected by endometrial cancer. PMID- 11487259 TI - Glomerular filtration rate prior to high-dose melphalan 200 mg/m(2) as a surrogate marker of outcome in patients with myeloma. AB - We correlated age and body surface area corrected glomerular filtration rate (GFR) at the time of high-dose melphalan (HDM) administration with treatment related toxicity (TT), time to disease progression and survival. Between 8/85 and 6/98, 144 newly diagnosed myeloma patients with a median age of 53 years (range, 31-72) received infusional chemotherapy with vincristine, doxorubicin and methylprednisolone, with/without cyclophosphamide or verapamil, followed by HDM 200 mg/m(2)and stem cell rescue. An additional patient received HDM at diagnosis. GFR was below normal in 38 patients (26%). At presentation, patients with low GFR at the time of HDM had higher serum creatinine, beta(2)M, stage III disease, calcium and Bence-Jones proteinuria. Toxic deaths post-HDM were similar in both groups (2/38 (5%) vs. 4/107 (4%)), though patients with low GFR had more oral mucositis (P< 0.0001), diarrhoea (P = 0.005) and infections (P = 0.04). The response and relapse rates of the 2 groups were not substantially different, but the median overall survival (OS) was significantly shorter in patients with low GFR (5.1 vs 7.5 years, P = 0.015). Multivariate analysis showed that a normal GFR and being in CR at the time of HDM were predictive of longer OS. We conclude that in context of high-dose chemotherapy for myeloma, dose of melphalan should not be modified in patients with low GFR and that early intensive treatment at relapse may improve results in patients with abnormal renal function. PMID- 11487261 TI - Haemophilus influenzae type b serology in childhood leukaemia: a case-control study. AB - Antibody to Haemophilus influenzae type b (Hib) polysaccharide (PRP) was measured in 42 children with acute lymphoblastic leukaemia (ALL) and 42 non-leukaemic hospital controls. Modelling anti-PRP concentrations as a function of age revealed that the slopes of the trend lines differed significantly between cases and controls (P = 0.05); anti-PRP concentrations were lower among younger cases, and higher among older cases, than among controls of the same ages. PMID- 11487262 TI - Oesophageal cancer in women: tobacco, alcohol, nutritional and hormonal factors. AB - We analysed 3 case-control studies from Italy and Switzerland including 114 women with squamous cell oesophageal cancer and 425 controls. The multivariate odds ratio was 4.5 for heavy smoking and 5.4 for heavy alcohol drinking. Fruit intake, vegetable intake, oral contraceptive and HRT use were inversely related to oesophageal cancer. PMID- 11487263 TI - Body weight and colorectal cancer risk in a cohort of Swedish women: relation varies by age and cancer site. AB - The relation between relative body weight and colorectal cancer among women is unclear. In a large prospective cohort study, we found a positive association only for distal cancers among younger women that became attenuated at older ages. These results support previous reports in which results were stratified by age or colorectal cancer site. PMID- 11487264 TI - South Asian ethnicity and material deprivation increase the risk of Epstein-Barr virus infection in childhood Hodgkin's disease. AB - In order to further define the factors associated with the observed variations in the Epstein-Barr virus-positive rate in childhood Hodgkin's disease, we have studied the effect of material deprivation (measured by the Townsend score) and ethnic origin on the frequency of Epstein-Barr virus-positivity in 55 cases of childhood Hodgkin's disease, diagnosed between 1981 and 1999, from a multi-ethnic region of the United Kingdom. Epstein-Barr virus status was determined by immunohistochemistry for the Epstein-Barr virus-encoded latent membrane protein 1. 62% of cases were Epstein-Barr virus-positive. Ethnic group was the strongest predictor of Epstein-Barr virus-positivity, with South Asians having a more than 20-fold risk of being Epstein-Barr virus-positive compared with non-South Asians. An increased risk was still present after adjusting for deprivation. Townsend scores were significantly higher (indicating more deprivation) in the Epstein Barr virus-positive group, particularly in males. The relative risk of Epstein Barr virus-positivity showed a gradient with increasing Townsend score; the risk being 7-times higher in the most deprived quartile compared with the least deprived group. Although the association between Townsend score and Epstein-Barr virus-positivity was reduced after adjusting for ethnic group, the risk of Epstein-Barr virus-positivity was still 3-times higher in the most deprived compared with the least deprived quartile. In addition, cases having 2 or more siblings were 5-times as likely to be Epstein-Barr virus-positive as those from smaller families. These results provide the first evidence of a strong association between Epstein-Barr virus-positive Hodgkin's disease and South Asian children from the United Kingdom. In addition, deprivation may increase the likelihood of Epstein-Barr virus-positive disease independently of ethnicity. PMID- 11487265 TI - Dietary fat, cholesterol and colorectal cancer in a prospective study. AB - The relationships between consumption of total fat, major dietary fatty acids, cholesterol, consumption of meat and eggs, and the incidence of colorectal cancers were studied in a cohort based on the Finnish Mobile Clinic Health Examination Survey. Baseline (1967-1972) information on habitual food consumption over the preceding year was collected from 9959 men and women free of diagnosed cancer. A total of 109 new colorectal cancer cases were ascertained late 1999. High cholesterol intake was associated with increased risk for colorectal cancers. The relative risk between the highest and lowest quartiles of dietary cholesterol was 3.26 (95% confidence interval 1.54-6.88) after adjusting for age, sex, body mass index, occupation, smoking, geographic region, energy intake and consumption of vegetables, fruits and cereals. Consumption of total fat and intake of saturated, monounsaturated, or polyunsaturated fatty acids were not significantly associated with colorectal cancer risk. Nonsignificant associations were found between consumption of meat and eggs and colorectal cancer risk. The results of the present study indicate that high cholesterol intake may increase colorectal cancer risk, but do not suggest the presence of significant effects of dietary fat intake on colorectal cancer incidence. PMID- 11487266 TI - Excess breast cancer risk and the role of parity, age at first childbirth and exposure to radiation in infancy. AB - Exposure to ionizing radiation is a known risk factor for breast cancer and the fertility pattern is a recognized modifier of breast cancer risk. The aim of this study was to elucidate the interaction between these 2 factors. This study is based on a Swedish cohort of 17 202 women who had been irradiated for skin haemangiomas in infancy between 1920 and 1965. The mean age at treatment was 6 months and the median breast dose was 0.05 Gy (range 0-35.8 Gy). Follow-up information on vital status, parity, age at first childbirth and breast cancer incidence was retrieved through record linkage with national population registers for the period 1958-1995. Analyses of excess relative risk (ERR) models were performed using Poisson regression methods. In this cohort, the fertility pattern differed from that in the Swedish population, with significantly fewer childbirths overall and before 25 years of age but more childbirth after that age. There were 307 breast cancers in the cohort and the standardized incidence ratio (SIR) was 1.22 (95% CI 1.09-1.36). A linear dose-response model with stratification for fertility pattern and menopausal status resulted in the best fit of the data. ERR/Gy was 0.33 (95% CI 0.17-0.53). In absolute terms this means an excess of 2.1 and 5.4 cases per Gy per 10(4) breast-years in the age groups 40 49 and 50-59 years respectively. The fertility pattern influenced the breast cancer risk in this irradiated population in a similar way to that observed in other studies. SIR at dose = 0 was highest, 2.31, among postmenopausal nulliparous women (95% CI 1.48-3.40, n = 62). SIR at dose = 0 was lowest in pre- or postmenopausal women with a first childbirth before 25 years of age; 0.89 (0.71-1.09) and 0.88 (0.58-1.25) respectively. Thus, in addition to the dose effect response in the cohort, part of the breast cancer excess could be explained by a different fertility pattern. The estimates of ERR/Gy for the various categories of age at first childbirth, number of children, menopausal status and ovarian dose were very similar, contradicting any interaction effects on the scale of relative risk. PMID- 11487267 TI - Long-term cancer patient survival achieved by the end of the 20th century: most up-to-date estimates from the nationwide Finnish cancer registry. AB - A new method of survival analysis, denoted period analysis, has recently been developed, which has been shown to provide more up-to-date estimates of long-term survival rates than traditional methods of survival analysis. We applied period analysis to data from the nationwide Finnish cancer registry to provide up-to date estimates of 5-, 10-, 15- and 20-year relative survival rates (RSR) achieved by the end of the 20th century. For most forms of cancer, period estimates of long-term survival are much higher than corresponding traditional survival estimates which suggests that for these cancers there has been ongoing major progress in survival rates in recent years which so far has remained undisclosed by traditional methods of survival analysis. For example, period analysis reveals that 10 year RSR have come close to (or even exceed) 80% for cancer of the corpus uteri and melanoma, 75% for breast cancer, 70% for bladder cancer, 65% for cancer of the cervix uteri, and 55% for cancer of the colon and prostate. Period analysis further reveals that 20 year RSR have now come close to (or even exceed) 75% for endometrial cancer and melanoma, 60% for breast cancer and cervical cancer, 55% for colon cancer and bladder cancer, and 40%-50% for cancer of the rectum, the ovaries, kidneys and nervous system. PMID- 11487268 TI - Population-based case-control study of soyfood intake and breast cancer risk in Shanghai. AB - We evaluated the association of soyfood intake and breast cancer risk in a population-based case-control study among Chinese women in Shanghai. Included in the study were 1459 cases and 1556 age-matched controls, with respective response rates of 91.1% and 90.3%. Usual soyfood intake was assessed using a food frequency questionnaire (FFQ). Separate analyses were performed for all subjects and for the subset who reported no recent change in soyfood intake. The intake levels of soyfoods among women in Shanghai are high, with 96.6% women reporting soyfood consumption at least once a week. A statistically non-significant reduced risk (odds ratio (OR) = 0.78 95% CI = 0.52-1.16) of breast cancer was observed among those who reported eating soyfood at least once a week. Compared to those in the lowest decile intake group, women in the highest decile intake group had a 30% reduced risk of breast cancer (OR = 0.66, 95% CI = 0.46-0.95), but no monotonic dose-response relation was observed (P for trend, 0.28). Stratified analyses showed that the inverse association was restricted primarily among women who had a high body mass index (BMI), with an adjusted OR of 0.30 (95% CI = 0.10 0.94) observed for the highest intake group. The reduction in risk was stronger for breast cancer positive for both oestrogen receptor (ER) and progesterone receptor (PR) (OR = 0.44, 95% CI = 0.25-0.78) than those with other ER/PR status. More pronounced inverse associations were observed in analyses among those who reported no recent change in soyfood intake than those conducted in all subjects. A dose-response relation between soyfood intake and breast cancer risk was observed in this subset of women (P for trend, 0.02), with an OR of 0.46 (95%CI = 0.28-0.75) for those in the highest decile intake group. No clear monotonic dose response relation was found between soyfood intake and breast cancer risk among regular soy eaters, but nevertheless the results suggest that regular soyfood consumption may reduce the risk of breast cancer, particularly for those positive for ER and PR; the effect may be modified by body mass index. PMID- 11487269 TI - High incidence of classic Kaposi's sarcoma in Mantua, Po Valley, Northern Italy (1989-1998). AB - The incidence of classic Kaposi's sarcoma was estimated in the province of Mantua, Po Valley, Northern Italy, yielding age-standardized rates of 2.5/100 000 men and 0.7/100 000 women (1989-98). Elevated rates in the rural zone of Viadana/Sabbioneta (5.0/100 000 men and 2.8/100 000 women) are among the highest so far reported for Italian communities. PMID- 11487270 TI - Upregulation and differential expression of matrilysin (MMP-7) and metalloelastase (MMP-12) and their inhibitors TIMP-1 and TIMP-3 in Barrett's oesophageal adenocarcinoma. AB - Oesophageal adenocarcinoma is believed to arise from metaplastic mucosa in the distal oesophagus, a condition also known as Barrett's oesophagus (BE). BE develops as a result of injury caused by refluxing gastric and duodenal contents and is associated with increased risk of malignant transformation. Matrix metalloproteinases (MMPs) have been implicated in all aspects of tumour progression; tumour growth, basement membrane degradation, invasion and metastatic spread. Using in situ hybridization, we investigated the expression patterns of collagenases-1 and -3, stromelysin-2, matrilysin, metalloelastase and TIMPs-1 and -3 in BE, adenocarcinoma and lymph-node metastases. Matrilysin was expressed abundantly in 12/15 tumours and in 4/6 lymph-node metastases and its expression correlated with the histological aggressiveness of tumour. Matrilysin and metalloelastase were upregulated already in BE. Stromelysin-2 and collagenase 3 expression was detected only in a few tumours. Collagenase-1 was expressed by cancer and stromal cells in 9/15 tumours. Tumour-infiltrating macrophages expressed metalloelastase in 13/15 cancers. TIMPs-1 and -3 were expressed in 12/15 and 11/15 tumours, respectively. Laminin-5 and tenascin were abundantly expressed at the invasive front of poorly differentiated tumours, but not in BE. Our results indicate that matrilysin is the principal MMP expressed by tumour cells in oesophageal adenocarcinoma, and further studies are needed to investigate whether matrilysin or tenascin-C could be used as a predictive marker for progression of BE to cancer. PMID- 11487271 TI - Expression and regulation of prostate androgen regulated transcript-1 (PART-1) and identification of differential expression in prostatic cancer. AB - Prostate androgen regulated transcript 1 (PART-1), is a gene predominantly expressed in the prostate gland and is regulated by androgens in human prostate cancer cell lines. Here, we report additional characteristics of PART-1 tissue expression and hormonal regulation and study its expression profile in human normal and matched prostate cancer tissues. Since PART-1 shows similarity to prostate-specific antigen (PSA) in prostate specificity and regulation, we hypothesized that it may be implicated in prostate carcinogenesis or may be a potential new biomarker. We used reverse transcriptase polymerase chain reaction (RT-PCR) to further characterize PART-1 tissue expression and hormonal regulation in the LNCaP prostate cancer cell line. RT-PCR analysis revealed that PART-1 is expressed not only in the prostate and salivary gland, but also in other tissues, including the thymus and placenta. In addition to androgen stimulation, PART-1 is also up-regulated by progestins, oestrogens and glucocorticoids. We further studied the expression of PART-1 in 27 paired (from the same patient) cancerous and non-cancerous prostatic tissues, with qualitative and quantitative RT-PCR (LightCycler technology), in order to examine whether PART-1 is overexpressed or underexpressed in cancer. Our results indicated that PART-1 is more frequently overexpressed in the cancerous prostatic tissue. We conclude that this gene is overexpressed in prostate cancer and may represent a novel prostate cancer tumour marker. PMID- 11487272 TI - HPV presence precedes abnormal cytology in women developing cervical cancer and signals false negative smears. AB - In a retrospective case-control study, we investigated high-risk HPV DNA presence by general primer GP5+/6+ PCR in the last normal cervical smear in the patient archives (i.e. baseline smear) of 57 women who later developed cervical cancer. Also, normal cervical smears of 114 age-matched control women were analysed. High risk HPV DNA was detected in 37 of the 57 (65%) baseline smears of the case women, and 7 (6%) of 114 smears of the control women (OR 28, 95% Cl 11-72). The HPV positive subsequent smears and cervical cancer biopsies of the case women contained the same HPV type as was detected in the baseline smear. After cytological revision, the baseline smears of 48 case women (84%) were reclassified as abnormal, 33 (69%) of which scored high-risk HPV DNA positive. Ultimately, an undisputable normal baseline smear was found in only 10 case women. In 7 (70%) of them this smear was HPV positive, whereas only 7 (7%) of 104 revised, undisputable normal smears of control women were high-risk HPV positive (OR 32, 95% Cl 6.8-153). The results showed that (1) high-risk HPV presence precedes abnormal cytology in women who develop cervical cancer, and (2) high risk HPV testing signals false-negative smears of women at risk of cervical cancer. PMID- 11487273 TI - c-KIT and c-KIT ligand (SCF) in synovial sarcoma (SS): an mRNA expression analysis in 23 cases. AB - In a previous immunophenotypic molecular-based analysis it was shown that bcl2 over-expression characterizes the SS gene profile in addition to the non-random translocations. Here we show that the over-expression of an additional potentially antiapoptotic gene, the c-KIT gene, is associated with this tumour. Interestingly, whereas bcl2 over-expression appears to be restricted to the spindle cell tumoral component, c-kit mainly involves the epithelial component of biphasic SS. Twenty-three primary and metastatic samples from 21 patients were analysed by immunophenotyping (23/23), immunoprecipitations and Western blotting (3/23), and RT-PCR (23/23). Ten cases were biphasic and 13 monophasic in sub type. Twelve, 10 and 1 case carried the SYT-SSX1, SYT-SSX2 and SYT-SSX4 fusion transcript, respectively. Co-presence of both c-Kit and SCF mRNA was observed in almost all cases (20/23), suggesting the occurrence of an autocrine loop. Immunophenotyping, confirmed by biochemical analyses, showed a modulation of c Kit expression which was faint in the spindle and strong in the epithelial component, respectively. The study was complemented by c-Met/HGF receptor/ligand expression and c-Met protein analysis with results superimposable to those already reported. Since in each tumour, epithelial and spindle cell components harbour the same type of translocation t(X;18) the present findings suggest a shifting of the anti-apoptotic role from BCL2 to c-KIT gene during the transition from the uncommitted spindle to the differentiated epithelial cells. PMID- 11487274 TI - Clinical significance of CDC25A and CDC25B expression in squamous cell carcinomas of the oesophagus. AB - CDC25A, CDC25B and CDC25C belong to a family of protein phosphatases which activate the cyclin-dependent kinase at different points of the cell cycle. According to accumulating evidence, CDC25A and CDC25B seem to possess oncogenic properties. We have analysed these expressions by immunohistochemistry, western blot and RT-PCR in a series of 100 patients with squamous cell carcinoma of the oesophagus. When compared with non-cancerous cells, CDC25A and CDC25B were strongly expressed in the cytoplasm of cancer cells, with positive (+) classification in 46% (46 cases) and 48% (48 cases), respectively. There was no significant correlation between CDC25A and CDC25B expression, nor was there any association with the expression of other cell cycle-regulating molecules, including cyclin D1, Rb, p16(INK4), p27(KIP1)and PCNA (proliferating cell nuclear antigen). CDC25A (+), as well as CDC25B (+), was more frequently found in patients with deeper tumour invasion and lymph node metastasis, while tumour size was correlated only with CDC25A expression. Postoperative survival was significantly poorer for CDC25A (+) patients than CDC25A (-) patients, but was not affected by the CDC25B status. Nuclear localization of CDC25A was observed in 51 cases (51%), regardless of its cytoplasmic expression, and was not associated with clinico-pathological factors or prognosis. Multivariate analysis revealed only the CDC25A status to be an independent significant prognostic factor among these biological and clinico-pathological factors. CDC25A but not CDC25B may be a new prognostic factor for squamous cell carcinoma of the oesophagus. Thus, regulation of the G1 checkpoint in the cell cycle may be important in oesophageal carcinogenesis, which may also involve many other oncogenes. PMID- 11487275 TI - CGH analysis of ductal carcinoma of the breast with basaloid/myoepithelial cell differentiation. AB - 2-18% of ductal carcinoma-No Special Type (NST) are reported to express basal cell keratin 14 and such tumours may have a different metastatic pattern and prognosis. We performed immunohistochemistry for cytokeratins 19 (luminal) and 14 (basal) on 92 ductal carcinoma-NST. Those tumours showing CK14 expression were further characterized by immunohistochemistry for myoepithelial cell phenotype and analysed by comparative genomic hybridization. The 7 cases of ductal carcinoma-NST exhibiting a basal cell phenotype were all grade III tumours and showed a molecular cytogenetic profile similar to more conventional myoepithelial cell carcinomas. Therefore it appears that grade III invasive ductal carcinomas contain a subset of tumours with specific morphological and cytogenetic characteristics, and probably prognosis for the patient. PMID- 11487276 TI - Reduced mammary gland carcinogenesis in transgenic mice expressing a growth hormone antagonist. AB - Several reports have provided evidence that body size early in life is positively correlated with risk of subsequent breast cancer, but the biological basis for this relationship is unclear. We examined tumour incidence in transgenic mice expressing a growth hormone (GH) antagonist and in non-transgenic littermates following exposure to dimethylbenz[ a ]anthracene (DMBA), a well characterized murine mammary gland carcinogen. The transgenic animals had lower IGF-I levels, were smaller in terms of body size and weight, and exhibited decreased tumour incidence relative to controls. The demonstration that both body size early in life and breast cancer incidence are influenced by experimental perturbation of the GH-IGF-I axis in a transgenic model provides evidence that variability between individuals with respect to these hormones underlies the relationship between body size early in life and breast cancer risk observed in epidemiological studies. PMID- 11487277 TI - In vivo evaluation of the early events associated with liver metastasis of circulating cancer cells. AB - The mechanism of metastasis formation remains still largely unknown. Many studies underline the importance and complexity of the initial arrest of the circulating tumour cells in the target organ, a key stage in metastasis occurrence. In our study, we evaluated by visual means the metastasis formation using an in vivo microscopy system in a murine model. Moreover, we investigated the involvement of P-selectin in these processes using immunohistochemistry and P-selectin knockout mice. The present study offers direct evidence of distinct pathways for tumour metastasis formation by a lymphoma cell - EL-4 and a solid tumour cell - C26. Off line analysis of the images and histological data confirmed that mechanical entrapment of the solid tumour cell, which had a bigger diameter than that of the liver sinusoids, promoted metastasis without any detectable involvement of adhesion molecules. On the other hand, we observed that lymphoma cells, in spite of their smaller diameter as compared to the sinusoids, promoted liver metastasis as well, but with the essential participation in their arrest of P-selectin, indicating an adhesion molecule-mediated pathway. PMID- 11487279 TI - The ability to accumulate deoxyuridine triphosphate and cellular response to thymidylate synthase (TS) inhibition. AB - Thymidylate synthase (TS) is an important enzyme catalysing the reductive methylation of dUMP to dTMP that is further metabolized to dTTP for DNA synthesis. Loss of viability following TS inhibition occurs as a consequence of depleted dTTP pools and at least in some cell lines, accumulation of dUTP and subsequent misincorporation of uracil into DNA. The expansion in dUTP pools is largely determined by the expression of the pyrophosphatase, dUTPase. Our previous work has shown that following TS inhibition the ability to accumulate dUTP was associated with an earlier growth inhibitory effect. 3 human lung tumour cell lines and HT29 human colon tumour cells transfected with dUTPase have been used to investigate the relationship between loss of viability following TS inhibition and dUTP accumulation. Cell cycle arrest typical of TS inhibition was an early event in all cell lines and occurred irrespective of the ability to accumulate dUTP or p53 function. However, a large expansion of dUTP pools was associated with mature DNA damage (4 h) and an earlier loss of viability following TS inhibition compared to cells in which dUTP pools were not expanded. In A549 cells damage to mature DNA may have been exacerbated by significantly higher activity of the excision repair enzyme, uracil-DNA glycosylase. Consistent with results using different inhibitors of TS, transfection of dUTPase into HT29 cells significantly reduced the cytotoxicity of a 24 h but not 48 h exposure to ZD9331. Although loss of viability can be mediated through dTTP deprivation alone, the uracil misincorporation pathway resulted in an earlier commitment to cell death. The relevance of this latter pathway in the clinical response to TS inhibitors deserves further investigation. PMID- 11487278 TI - Impact of thymidine phosphorylase surexpression on fluoropyrimidine activity and on tumour angiogenesis. AB - Tumoral thymidine phosphorylase (TP) appears to play a dual role by being involved in neoangiogenesis and by activating 5FU prodrugs at the tumoral target site. The aim of the study was to investigate more thoroughly these potential physiological and pharmacological roles of TP. A rat carcinoma cell line (PROb) was transfected with TP/PD-ECGF in order to study the effect of the overexpression of this enzyme (1) on the sensitivity of cells to 5'DFUR and 5FU in vitro and (2) on tumour growth in vivo by using a syngenic tumour model in the BDIX rat (hepatic tumours, sub-cutaneous tumours). Cytotoxic effects of 5'DFUR, and to a lesser extent those of 5FU, were enhanced in TP clones as compared to control cells: there was a highly significant correlation between TP activity and in vitro sensitivity to 5'DFUR (r2= 0.91, P = 0.0002, n = 8) and, to a lesser extent, to 5FU (r2= 0.49, P = 0.053, n = 8). The impact of TP transfection on tumour growth was relatively modest and concerned only the initial stages of tumour expansion. Staining of TP tumours for endothelial (factor VIII) cells was always higher than controls. The staining ratio (TP/controls) tended to be reduced as tumours increased in size. The stability of TP expression was checked both in vitro (TP activity measurement) and in vivo (RT-PCR determinations) and there was no loss of TP expression over time which could be advanced to explain the progressive weakening of the impact of TP overexpression on both tumour growth and neoangiogenesis. PMID- 11487280 TI - Antagonists of retinoic acid receptors (RARs) are potent growth inhibitors of prostate carcinoma cells. AB - Novel synthetic antagonists of retinoic acid receptors (RARs) have been developed. To avoid interference by serum retinoids when testing these compounds, we established serum-free grown sub-lines (>3 years) of the prostate carcinoma lines LNCaP, PC3 and DU145. A high affinity pan-RAR antagonist (AGN194310, K(d) for binding to RARs = 2-5 nM) inhibited colony formation (by 50%) by all three lines at 16-34 nM, and led to a transient accumulation of flask-cultured cells in G1 followed by apoptosis. AGN194310 is 12-22 fold more potent than all-trans retinoic acid (ATRA) against cell lines and also more potent in inhibiting the growth of primary prostate carcinoma cells. PC3 and DU145 cells do not express RARbeta, and an antagonist with predominant activity at RARbeta and RARgamma (AGN194431) inhibited colony formation at concentrations (approximately 100 nM) commensurate with a K(d)value of 70 nM at RARgamma. An RARalpha antagonist (AGN194301) was less potent (IC(50) approximately 200 nM), but was more active than specific agonists of RARalpha and of betagamma. A component(s) of serum and of LNCaP-conditioned medium diminishes the activity of antagonists: this factor is not the most likely candidates IGF-1 and EGF. In vitro studies of RAR antagonists together with data from RAR-null mice lead to the hypothesis that RARgamma-regulated gene transcription is necessary for the survival and maintenance of prostate epithelium. The increased potencies of RAR antagonists, as compared with agonists, suggest that antagonists may be useful in the treatment of prostate carcinoma. PMID- 11487282 TI - Cross-modality temporal processing deficits in developmental phonological dyslexics. AB - Neuroanatomical evidence suggests that poor readers may have abnormal lateral (LGN) and medial (MGN) geniculate nuclei responsible for temporal processing in visual and auditory domains respectively (Livingstone & Galaburda, 1993). Although behavioral evidence does support this neuroanatomical evidence in that poor readers have performed poorly on visual and auditory tasks thought to require the utilization of the LGN and MGN, respectively, appropriate examination of the coexistence of these behavioral abnormalities in the same population of poor readers has yet to take place. The present study examined correlations between visual and auditory temporal processing scores of all readers (collapsed groups), good readers, and poor readers who were isolated into phonological and surface dyslexic subtypes. The same subjects and data from Cestnick and Coltheart (1999) and Cestnick and Jerger (2000) were used to run the analyses. Results demonstrated a multitude of correlations between these tasks for the phonological dyslexic group only. It is contended that cross-modality temporal processing deficits may exist in poor nonlexical (phonological dyslexics) as opposed to poor lexical (surface dyslexics) readers. It is conceivable that phonological dyslexics may also have deficiencies within the LGN and MGN, or perhaps within systems related to these nuclei. The precise cause of these processing patterns and correlations is still unknown. PMID- 11487281 TI - Chimaeric Lym-1 monoclonal antibody-mediated cytolysis by neutrophils from G-CSF treated patients: stimulation by GM-CSF and role of Fc gamma -receptors. AB - Chimaeric Lym-1 (chLym-1) is a monoclonal antibody generated by fusing the variable region genes of murine Lym-1 to human gamma1 and kappa constant regions. Owing to its selectivity and avidity for human malignant B cells, it is an attractive candidate for developing immune-interventions in B-lymphomas. In the attempt to identify rational bases for optimizing potential chLym-1 related therapeutic approaches, we studied the ability of this ch-mAb to trigger neutrophil-mediated Raji cell cytolysis in cooperation with two neutrophil related cytokines, G-CSF and GM-CSF. ChLym-1 triggered low levels of cytolysis by normal neutrophils but induced consistent cytolysis in neutrophils from individuals treated with G-CSF. When exposed to GM-CSF, neutrophils from subjects treated with G-CSF became potent effectors, also leading to 75% lysis. By using mAbs specific for distinct FcgammaRs, normal neutrophils were inhibited by mAb IV.3, suggesting the intervention of FcgammaRII, constitutively expressed on the cells. On the other hand, neutrophils from patients treated with G-CSF were inhibited by mAb IV.3 plus mAb 197, a finding consistent with a cooperative intervention of FCgammaRII and G-CSF-induced FcgammaRI. The anti-FcgammaRIII mAb 3G8 promoted significant enhancement of the neutrophil cytolytic efficiency. Therefore, neutrophil FcgammaRIII behaves as a down-regulator of the cytolytic potential. The present findings suggest new attempts to develop mAb-based and G CSF/GM-CSF combined immune-interventions in B lymphomas. PMID- 11487283 TI - Test of a potential link between analytic and nonanalytic category learning and automatic, effortful processing. AB - The link between automatic and effortful processing and nonanalytic and analytic category learning was evaluated in a sample of 29 college undergraduates using declarative memory, semantic category search, and pseudoword categorization tasks. Automatic and effortful processing measures were hypothesized to be associated with nonanalytic and analytic categorization, respectively. Results suggested that contrary to prediction strong criterion-attribute (analytic) responding on the pseudoword categorization task was associated with strong automatic, implicit memory encoding of frequency-of-occurrence information. Data are discussed in terms of the possibility that criterion-attribute category knowledge, once established, may be expressed with few attentional resources. The data indicate that attention resource requirements, even for the same stimuli and task, vary depending on the category rule system utilized. Also, the automaticity emerging from familiarity with analytic category exemplars is very different from the automaticity arising from extensive practice on a semantic category search task. The data do not support any simple mapping of analytic and nonanalytic forms of category learning onto the automatic and effortful processing dichotomy and challenge simple models of brain asymmetries for such procedures. PMID- 11487284 TI - Visual signal detection measured by event-related potentials. AB - Signal Detection tasks typically involve within-subject signal changes. Such a procedure does not lend itself to event-related potential (ERP) experiments where the need for averaging necessitates the maintenance of consistent stimulus parameters. In the present ERP study we adopt a novel approach to thresholding that allows within-subject signal manipulation. The Signal Detection task required the identification of letter targets, formed from dots, in a random dot field. ERP waveforms were segmented into three windows corresponding to N1, N2, and P300 components. Analysis shows that ERP variations are dependent on both task demands and response characteristics for N1, N2, and P300 components. PMID- 11487285 TI - A neuropsychological study of dissociation in cortical and subcortical functioning in obsessive-compulsive disorder by Tower of Hanoi task. AB - Several biological models of Obsessive-Compulsive Disorder (OCD) have focused on the roles frontal cortex and basal ganglia dysfunctions play in the expression of the disorder. From a neuropsychological point of view, previous reports have underlined the possible involvement of the prefrontal cortex in declarative functions and the basal ganglia in procedural ones. A possible dissociation of cortical and subcortical functioning has been studied using the Hanoi Tower Task to explore different neuropsychological aspects of problem-solving procedures. Our results indicate that differential cortical and subcortical dysfunctions could contribute to OCD pathophysiology and that procedural and declarative forms might be independent of each other. PMID- 11487286 TI - Renatus renatus: the Cartesian tradition in British neuroscience and the neurophilosophy of John Carew Eccles. AB - J. C. Eccles (1903-1997) had a highly distinguished career in neurophysiology, being awarded the Nobel Prize for Medicine or Physiology in 1963. This paper sets him within the Cartesian tradition of British neurophysiology initiated by Thomas Henry Huxley in the mid-19th century. It shows how the mind-brain problematique of the Cartesian tradition troubled him throughout his career, leading him finally to a solution in terms of quantum microphysics and microphysiology. This position, which has subsequently become fashionable, is discussed and shown (at least in the form Eccles espoused) to provide no solution to the problem posed by Descartes in the early 17th century. PMID- 11487287 TI - Shared processes in spatial rotation and musical permutation. AB - An experiment was conducted in which subjects performed a three-dimensional spatial rotation test (24 trials) and a new test involving judgments of musical permutations (64 trials). Two types of musical permutations were used, including retrograde and inverse. In a retrograde permutation, the criterion melody was played backward in the test melody, and in an inverse permutation, an ascending or descending interval in the criterion melody became an opposite in the test melody. Subjects included 32 male and 64 female undergraduates at the University of Toronto. Regression analysis clearly showed that it was easiest to compare short retrograde permutations and that accuracy at discerning retrograde permutations predicted accuracy at judging spatial rotations. The implication is that a higher order ability to discriminate contour underlies both kinds of judgments. PMID- 11487288 TI - Does aging affect the allocation of visual attention in global and local information processing? AB - The earliest studies of Navon (1977) showed that in certain conditions the visual perception of hierarchical stimuli generates a global precedence effect. This hypothesis is based on the joint occurrence of two effects: a response time (RT) advantage for identifying global targets and interference by global distractors when responding to a local target. The aim of the present study was to determine the involvement of attentional mechanisms and whether certain aspects are specifically altered by normal aging. In a selective-attention task, a complete global precedence effect was found for young and old subjects. However, for the old subjects, global interference on local identification was more pronounced than for the young subjects. In a divided-attention task, the RT advantage was affected by attention shifts between global and local forms for both young and old subjects, but the global interference effect did not change. PMID- 11487289 TI - Reliability and magnitude of auditory laterality effects: the influence of attention. AB - Three experiments investigated the effect of attention on the reliability and magnitude of laterality effects in a dichotic listening task. In Experiment 1, 40 undergraduate students were randomly assigned to either a free-recall or focused attention condition. In Experiment 2, 40 undergraduate students completed a dichotic listening task with exogenous cueing. They were randomly assigned to either a 150-ms stimulus onset asynchrony (SOA) or a 450-ms SOA condition. In Experiment 3, 20 participants completed a task where the SOA for the exogenous cue was randomized on a trial to trial basis. Results indicated that focused attention increased the magnitude of the laterality effect. Contrary to predictions, this finding was not due to reduced variability in the focused attention task compared to the free-recall task. In addition, a cueing tone was only effective at directing attention in Experiment 3. Specifically, a significant right ear advantage observed at the 150-ms SOA was reduced at the 450 ms SOA. It appears that, in Experiment 3, the tone was effective at controlling attention because it reduced the systematic bias that has been suggested to account for the laterality effects observed in dichotic tasks. PMID- 11487290 TI - Role of individual neurons and neural networks in cognitive functioning of the brain: a new insight. AB - The prevailing concept in modern neuroscience is that neuron networks play a dominant role in the functioning of the nervous system, whereas the role of individual neurons is rather insignificant. This concept suggests that "individuality" of single neurons is primarily determined by their place in a network rather than their intrinsic properties. Here I argue that individual neurons may play an important, if not decisive, role in performing cognitive functions of the brain. This tentative viewpoint is supported by experimental and clinical insights into disorders of cognitive functions and by genetic studies of cognitive abilities and disabilities. The results obtained in these studies indicate that many specific cognitive functions are carried out by groups of highly specialized neurons whose roles in performing these functions are genetically predetermined and their activity could not be substituted by the activity of other neurons. In this context, the main role of neural networks and intercellular interactions is to form dynamic ensembles of neurons involved in performing a given cognitive function. PMID- 11487292 TI - Does language shape thought? Mandarin and English speakers' conceptions of time. AB - Does the language you speak affect how you think about the world? This question is taken up in three experiments. English and Mandarin talk about time differently--English predominantly talks about time as if it were horizontal, while Mandarin also commonly describes time as vertical. This difference between the two languages is reflected in the way their speakers think about time. In one study, Mandarin speakers tended to think about time vertically even when they were thinking for English (Mandarin speakers were faster to confirm that March comes earlier than April if they had just seen a vertical array of objects than if they had just seen a horizontal array, and the reverse was true for English speakers). Another study showed that the extent to which Mandarin-English bilinguals think about time vertically is related to how old they were when they first began to learn English. In another experiment native English speakers were taught to talk about time using vertical spatial terms in a way similar to Mandarin. On a subsequent test, this group of English speakers showed the same bias to think about time vertically as was observed with Mandarin speakers. It is concluded that (1) language is a powerful tool in shaping thought about abstract domains and (2) one's native language plays an important role in shaping habitual thought (e.g., how one tends to think about time) but does not entirely determine one's thinking in the strong Whorfian sense. PMID- 11487293 TI - Attentional limitations in the sensing of motion direction. AB - Attentional constraints in the perceptual analysis of motion direction were examined using two independent paradigms: redundant target visual search and the analysis of fluctuations in discrimination accuracy at threshold. Results from both methods implied that directions of object motion are analyzed in parallel when those motions are translations, independent of the observer's line of sight. The registration of rotation direction appears to be subject to a qualitatively different protocol, one that is highly capacity limited and serial-like. These results suggest that scene-based descriptions, as opposed to image-based descriptions of motion, mediate the allocation of attention. PMID- 11487294 TI - The development of ordinal numerical competence in young children. AB - Two experiments assessed ordinal numerical knowledge in 2- and 3-year-old children and investigated the relationship between ordinal and verbal numerical knowledge. Children were trained on a 1 vs 2 comparison and then tested with novel numerosities. Stimuli consisted of two trays, each containing a different number of boxes. In Experiment 1, box size was held constant. In Experiment 2, box size was varied such that cumulative surface area was unrelated to number. Results show children as young as 2 years of age make purely numerical discriminations and represent ordinal relations between numerosities as large as 6. Children who lacked any verbal numerical knowledge could not make ordinal judgments. However, once children possessed minimal verbal numerical competence, further knowledge was entirely unrelated to ordinal competence. Number may become a salient dimension as children begin to learn to count. An analog magnitude representation of number may underlie success on the ordinal task. PMID- 11487295 TI - Specific soft tissue mobilization in the management of soft tissue dysfunction. AB - SUMMARY. Following injury, the ability of soft tissue to tolerate the demands of functional loading decreases. A major part of the management of soft tissue dysfunction lies in promoting soft tissue adaptation to restore the tissue's ability to cope with functional loading. Specific soft tissue mobilization (SSTM) uses specific, graded and progressive application of force by the use of physiological, accessory or combined techniques either to promote collagen synthesis, orientation and bonding in the early stages of the healing process, or to promote changes in the viscoelastic response of the tissue in the later stages of healing. SSTM should be applied in combination with rehabilitation regimes to restore the kinetic control of the tissue. The principles of SSTM are reviewed with regard to the general principles of treating soft tissue dysfunction and areas identified for further research into this field. Copyright 1998 Harcourt Publishers Ltd. PMID- 11487296 TI - Insufficient lumbopelvic stability: a clinical, anatomical and biomechanical approach to 'a-specific' low back pain. AB - SUMMARY. A clinical, anatomical and biomechanical model is introduced based on the concept that under postural load specific ligament and muscle forces are necessary to intrinsically stabilize the pelvis. Since load transfer from spine to pelvis passes through the sacroiliac (SI) joints, effective stabilization of these joints is essential. The stabilization of the SI joint can be increased in two ways. Firstly, by interlocking of the ridges and grooves on the joint surfaces (form closure); secondly, by compressive forces of structures like muscles, ligaments and fascia (force closure). Muscle weakness and insufficient tension of ligaments can lead to diminished compression, influencing load transfer negatively. Continuous strain of pelvic ligaments can be a consequence leading to pain. For treatment purposes stabilization techniques followed by specific muscle strengthening procedures are indicated. When there is a loss of force closure, for instance in peripartum pelvic instability, application of a pelvic belt can be advised. Copyright 1998 Harcourt Publishers Ltd. PMID- 11487297 TI - Effect of position on the posteroanterior stiffness of the lumbar spine. AB - SUMMARY. Physiotherapists commonly use motion testing techniques in the assessment of patients with spinal disorders. One of the techniques considered important in the assessment of low back pain is the application of posteroanterior (PA) forces to the lumbar spinous processes to evaluate spinal stiffness and symptom response. The purpose of this study was to investigate the effect of spinal position on the measured lumbar PA stiffness. The PA stiffness at the L3 and L5 vertebral levels was assessed in 12 painfree subjects in three prone lying positions (flexion, neutral and extension). PA forces were applied to the spinous processes, under quasi-static loading conditions, using a specifically built apparatus. The stiffness coefficient was calculated from the slope of the regression line fitted to the truncated (35N-80N) force-displacement curves. At both vertebral levels, the PA stiffness was position dependent, with the lowest values obtained in the neutral position. The mean stiffness increased by 12.4% in extension and 31.9% in flexion. In all positions PA stiffness was significantly greater at L5 than at L3. These results suggest that clinical assessments of lumbar PA stiffness may be influenced by the patient position in which the test is performed. Comparative assessments of PA stiffness over time should be performed with the spine supported in the same position. Copyright 1998 Harcourt Publishers Ltd. PMID- 11487298 TI - Reliability of the OSI CA 6000 Spine Motion Analyzer with a new skin fixation system when used on the thoracic spine. AB - SUMMARY. Following the independent development in the University of Brighton of a new skin fixation system for use with the OSI CA 6000 Spine Motion Analyzer (SMA), a series of studies was initiated to test the reliability of the instrument with the new skin fixation system, and the repeatability of the results thus obtained. Successful completion of previous repeatability and reliability studies using the SMA to measure lumbar spine motion, led to the development of a repeatability study, utilizing the instrument on the thoracic spine. This involved 11 asymptomatic subjects and three separate operators. The mean ranges of motion obtained from these results are reported and analyses of variance were used to calculate Intra-class Correlation Coefficients (ICCs) as an indication of reliability and repeatability. These are summarized as follows: Flexion/Extension ranges 0.77; lateral flexion ranges 0.88; rotation ranges 0.84. It is concluded that the instrument is capable of producing repeatable results when used by suitably trained clinicians and the results reported justify progression to the collection of normative data and other related studies for thoracic spine motion, utilizing the CA 6000 Spine Motion Analyzer with the new skin fixation system. Copyright 1998 Harcourt Publishers Ltd. PMID- 11487299 TI - Comparison of two passive mobilizing techniques following Colles' fracture: a multi-element design. AB - SUMMARY. This study compared the outcomes of using two passive mobilizing techniques, passive sustained stretches and oscillations, to treat post immobilization sequelae of Colles' fractures types I and III. The study used a type of single subject, multi-element design, to enable comparisons of the two techniques in the same subject, when rapid change and limited reversibility was expected and research had to occur concurrently with treatment. Eight female subjects were randomly assigned to predesigned sets of treatment conditions for six consecutive sessions. Each subject had completed the usual 6 weeks of immobilization of their affected arm within 3 days of commencing the study. Results indicate that both passive mobilizing techniques increased the range of wrist extension, but, if pain was present, oscillations were more effective. Consistent with this, sustained stretches were more effective in the absence of pain or later in a series of treatments. Copyright 2000 Harcourt Publishers Ltd. PMID- 11487300 TI - Mobilizing the nervous system in cervical cord compression. PMID- 11487301 TI - Plasma fatty acid levels in autistic children. AB - Phospholipid fatty acids are major structural components of neuronal cell membranes, which modulate membrane fluidity and hence function. Evidence from clinical and biochemical sources have indicated changes in the metabolism of fatty acids in several psychiatric disorders. We examined the phospholipid fatty acids in the plasma of a population of autistic subjects compared to mentally retarded controls. Our results showed a marked reduction in the levels of 22: 6n 3 (23%) in the autistic subjects, resulting in significantly lower levels of total (n-3) polyunsaturated fatty acids (PUFA) (20%), without significant reduction in the (n-6) PUFA series, and consequently a significant increase in the (n-6)/(n-3) ratio (25%). These variations are discussed in terms of potential differences in PUFA dietary intake, metabolism, or incorporation into cellular membranes between the two groups of subjects. These results open up interesting perspectives for the investigation of new biological indices in autism. Moreover, this might have new therapeutic implications in terms of child nutrition. PMID- 11487302 TI - Bradykinin and electrical stimulation increase prostaglandin production in the rat vas deferens. AB - The epididymal portion of the rat vas deferens produced prostaglandins (PG) E(2), F(2alpha)and 6-keto F(1alpha). Electrical stimulation (ES, 0.1 Hz, 1 ms) increased such production by 100%, and similar results were obtained in the presence of 1.0 microM bradykinin (Bk). When both stimuli were applied simultaneously, the increases in PG production were 1100% for PGE(2), 800% for PGF(2alpha)and 400% for PG6-keto F(1alpha). Prazosin abolished the effect of ES on PG production. A selective Bk B(2)-receptor antagonist abolished the increase in PG production induced by Bk, both in non-stimulated and in ES tissues. Bk (1.0 microM) elicited contractile responses in non-stimulated as well as in ES tissues, responses that were not modified in the presence of 10 microM indomethacin. In conclusion, the effects of Bk on prostaglandin production appears to depend on the activation of B(2) receptors, while the increase in prostaglandin release induced by ES, and the effects observed with both stimuli simultaneously, should be mediated by the release of noradrenaline and the subsequent activation of alpha(1) adrenoceptors. PMID- 11487303 TI - Profile of eicosanoids produced by human saphenous vein endothelial cells and the effect of dietary fatty acids. AB - Human saphenous vein endothelial cells (HSVECs) derived from primary cultures of adult human veins constitute an excellent in vitro model for studying human endothelial metabolism. In this study we report the (14)C-labelled prostanoid profile of HSVECs under resting and stimulated conditions and the effect of the n 3 polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid on them. Results indicate that HSVECs while under resting conditions produce mainly prostaglandin F(2alpha)(PGF(2alpha)). After stimulation with calcium ionophore A23187, the cells were found to synthesise PGI(2), PGE(2)and PGF(2alpha) as major products and thromboxane B(2)and PGD(2)as minor products. Production of (14)C labelled hydroxyeicosatetraenoic acids was not detected. Eicosapentaenoic acid was found to inhibit basal and stimulated prostanoid production whereas docosahexaenoic acid inhibited basal but strongly increased stimulated prostanoid production. These results may offer the basis for further studies aiming to investigate targets for pharmacological intervention in inflammatory conditions. PMID- 11487304 TI - Effects of conjugated linoleic acids and docosahexaenoic acid on rat liver and reproductive tissue fatty acids, prostaglandins and matrix metalloproteinase production. AB - Long chain n-6 and n-3 fatty acids play important roles in labor and delivery. These effects may be mediated by prostaglandin (PG) synthesis and by regulation of matrix metalloproteinases (MMPs), both of which play roles in uterine contraction, cervical ripening and rupture of fetal membranes. The effects of altering dietary n-6:n-3 long chain fatty acid ratios, and the addition of dietary conjugated linoleic acids (CLA) and docosahexaenoic acid (DHA) on fatty acid composition of reproductive tissues, PG synthesis in liver and reproductive tissue and serum MMP levels were examined in pregnant rats. Modified AIN-96G diets with n-6:n-3 ratios of 7:1 and 34:1 with and without added 1.1% (by weight) conjugated linoleic acid (CLA) and/or 0.3% (by weight) DHA were fed through day 20 of gestation. Reproductive tissues readily incorporated both DHA and CLA. CLA significantly (P<0.05) depressed PGF(2 alpha)synthesis in placenta, uterus and liver by 50% when the n-6:n-3 ratio was 7:1 and by 66% at 34:1 ratio. Significant differences (P<0.05) in PGE(2)synthesis in uterus and liver were seen only between groups fed the high ratio of n-6:n-3 without CLA, and the low ratio with CLA. Addition of CLA to DHA containing diets depressed PGF(2alpha) by one-third in uterus and liver (P<0.05). Serum MMP-9 and active MMP-2 were suppressed (P<0.05) by addition of either CLA or DHA. PMID- 11487306 TI - Essential fatty acids as possible mediators of the actions of statins. AB - Statins and polyunsaturated fatty acids have similar actions: both enhance endothelial nitric oxide synthesis, inhibit the production of pro-inflammatory cytokines, lower cholesterol levels, prevent atherosclerosis and are of benefit in coronary heart disease, stroke and osteoporosis. Statins enhance the conversion of linoleic acid and eicosapentaenoic acid to their long chain derivatives. Animals with essential fatty acid deficiency show an increase in HMG CoA reductase activity, which reverts to normalcy following topical application of linoleic acid. Similarly to statins, polyunsaturated fatty acids also inhibit HMG-CoA reductase activity. In view of the similarity in their actions and as statins influence essential fatty acid metabolism, it is suggested that essential fatty acids and their metabolites may serve as second messengers of the actions of statins. PMID- 11487305 TI - Stimulated release of arachidonic acid from rat liver cells by celecoxib and indomethacin. AB - The selective cyclooxygenase-2 (COX) inhibitor, celecoxib, and the non-selective COX-1 and COX-2 inhibitor, indomethacin, at microM concentrations, stimulate the release of arachidonic acid from rat liver cells (the C-9 cell line). Other non steroidal anti-inflammatory drugs were tested, including salicylic acid, acetylsalicylic acid, sulindac sulfone, sulindac sulfide, ibuprofen, piroxicam and the selective COX-2 inhibitor, rofecoxib, and the analgesic, acetaminophen. These compounds do not stimulate the release of arachidonic acid. The stimulated release by celecoxib is not affected by dexamethasone or the translation and transcription inhibitors, actinomycin and cycloheximide. The increased arachidonic acid release may be associated with induction of apoptosis. PMID- 11487307 TI - Effect of a cysteinyl leukotriene antagonist, pranlukast hydrate, on acetaldehyde induced bronchoconstriction in asthmatic patients. AB - Acetaldehyde is a main factor of alcohol-induced asthma. We previously reported that the cysteinyl leukotriene (cys-LT) receptor antagonist, pranlukast hydrate, inhibits acetaldehyde-induced airway hyperresponsiveness in guinea pigs. The purpose of this study was to evaluate the involvement of cys-LT on bronchial responsiveness to acetaldehyde in asthmatic patients. We investigated the bronchial response to inhalation of acetaldehyde in 10 asthmatic patients, who were treated with placebo or pranlukast hydrate (225.5 mg), a cys-LT receptor antagonist, twice a day for 1 wk using a double-blind, randomized, placebo controlled, cross-over design. Although a remarkable improvement of acetaldehyde bronchoconstriction was observed in 3 out of 10 subjects, PC(20)-AcCHO values were identical between placebo [12.0 (GSEM, 1.192) mg/ml] and pranlukast [14.7 (GSEM, 1.245) mg/ml] groups. The changes in bronchial responsiveness to acetaldehyde were similar in the six patients who had never experienced alcohol induced asthma and the four who had. In conclusion, cys-LTs are not involved in acetaldehyde-induced bronchoconstriction. PMID- 11487308 TI - Regular ingestion of opuntia robusta lowers oxidation injury. AB - The influence of opuntia robusta (prickly pear), a traditionally used dietary nutrient against diabetes mellitus among the American Indian population, was examined in 15 young patients suffering from familial heterozygous isolated hypercholesterolemia. Oxidation injury was determined via 8-epi-PGF(2 alpha)in plasma, serum and urine. Daily consumption of 250 g broiled edible pulp of prickly pear had no influence on body weight and body fat composition. Total cholesterol was lowered (P<0.01) as was LDL-cholesterol (P<0.04). No significant changes were observed either in triglycerides or in HDL. Prickly pear induced a significant decrease in plasma (27.9+/-3.3-->25.6+/-3.2;P<0.03), serum (302.0+/ 11.4-->283.2+/-14.5;P<0.0003) and urinary (355.9+/-18.4-->323.9+/-16;P<0.00002) 8 epi-PGF(2alpha)values. The findings on a decrease of 8-epi-PGF(2alpha)were more pronounced in females than in males, the highest significance being found in urine, while, in contrast, the effects on total- and LDL-cholesterol were more pronounced in males. A prerunning 4 weeks period of dietary counseling had no significant effect on either of the parameters examined. These findings indicate that the regular ingestion of opuntia robusta is able to significantly reduce in vivo oxidation injury in a group of patients suffering from familial hypercholesterolemia. This traditional food of the American Indians thus may have a significant cardiovascular benefit. PMID- 11487309 TI - Effect of three low-dose fish oil supplements, administered during pregnancy, on neonatal long-chain polyunsaturated fatty acid status at birth. AB - Adequate long-chain polyunsaturated fatty acid (LCP) status during pregnancy is important. We studied the effect of three low-dose fish oil supplements, administered during uncomplicated pregnancy, on neonatal LCP status at term delivery. Supplements were administered from the second trimester to delivery, either as fish oil capsules ("fish-1": 336 mg LCPomega3, n=15; and "fish-3": 1,008 mg LCPomega3, n=20) or milk-based supplement ("Mum": 528 mg LCPomega3, n=24). Fifty-seven untreated women served as controls. Fatty acids of umbilical veins (UV) and arteries (UA) were measured. The fish-1 group showed no differences, compared to controls. The Mum group had higher 20:5omega3, 22:5omega3, 22:6omega3, LCPomega3 and 22:6omega3/22:5omega6 in UV and UA. The fish-3 group had higher 22:5omega3 and 22:6omega3 (UA), LCPomega3 and 22:6omega3/22:5omega6 (UV and UA) and 20:3omega6 (UV). A 500-1000 mg daily LCPomega3 supplement, taken either as a milk-based supplement or fish oil capsules, effectively increases fetal LCPomega3 status, without affecting LCPomega6 status. PMID- 11487310 TI - Ligand dependence of metal-metal bonding in the d(3)d(3) dimers M(2)X(9)(n-) (M(III) = Cr, Mo, W; M(IV) = Mn, Tc, Re; X = F, Cl, Br, I). AB - The ligand dependence of metal-metal bonding in the d(3)d(3) face-shared M(2)X(9)(n-) (M(III) = Cr, Mo, W; M(IV) = Mn, Tc, Re; X = F, Cl, Br, I) dimers has been investigated using density functional theory. In general, significant differences in metal-metal bonding are observed between the fluoride and chloride complexes involving the same metal ion, whereas less dramatic changes occur between the bromide and iodide complexes and minimal differences between the chloride and bromide complexes. For M = Mo, Tc, and Re, change in the halide from F to I results in weaker metal-metal bonding corresponding to a shift from either the triple metal-metal bonded to single bonded case or from the latter to a nonbonded structure. A fragment analysis performed on M(2)X(9)(3-) (M = Mo, W) allowed determination of the metal-metal and metal-bridge contributions to the total bonding energy in the dimer. As the halide changes from F to I, there is a systematic reduction in the total interaction energy of the fragments which can be traced to a progressive destabilization of the metal-bridge interaction because of weaker M-X(bridge) bonding as fluoride is replaced by its heavier congeners. In contrast, the metal-metal interaction remains essentially constant with change in the halide. PMID- 11487311 TI - Bimetallic reactivity. One-site addition two-metal oxidation reactions using a di Co(II) complex of a binucleating ligand with 5- and 6-coordinate sites. AB - The preparation of an unsymmetrical binucleating ligand bearing a bridging oxadiazole ring flanked on one side by three ligands and on the other by four ligands is described. When bound to two metals, the ligand forms complexes where the metals are in 5- and 6-coordinate sites after the incorporation of an exogenous bridging ligand. A di-Co(2+) complex of this ligand has been prepared containing a hydroxide bridge. The complex is readily oxidized to the di-Co(3+) state by outer sphere electron transfer with ferrocenium ions. Addition of Br(2) or NO(2)(+) to the di-Co(2+) complex leads to the rapid formation of the di Co(3+) bromo or nitro complexes, respectively. The ligand characteristics which allow for double oxidation with ferrocenium ions and for the one-site addition two-metal oxidations with Br(2) and NO(2)(+) are discussed in terms of mechanical coupling between the two metal sites. PMID- 11487312 TI - Weak M(II)-azide-4,4'-bipy ferromagnets based on unusual diamondoid (M = Mn) and 2D arrays (M = Co, Ni). AB - Four compounds of general formula [M(4,4'bipy)(N(3))2](n) (M = Mn (1), Zn (2), Co (3), Ni (4)) have been synthesized and magnetostructurally characterized by means of X-ray diffraction analysis, IR and ESR spectroscopies, and measurements of the magnetic susceptibility and magnetization. Compound 1 (C(10)H(8)N(8)Mn) crystallizes in the tetragonal P4(3)2(1)2 space group, Z = 4, with a = 8.229(2), b = 8.229(2), and c = 16.915(2) A. It exhibits an acentric 3D structure where Mn(II) ions are linked through EE-azide groups resulting in a diamondoid network. The 4,4'bipy ligands are coordinated on the axial positions of the octahedral spheres reinforcing the intermetallic connections. Weak ferromagnetism arising from spin canting is observed for compound 1. Compounds 2, 3, and 4 are proposed to be isomorphous and would consist of a 2D array where alternating EO + EE/EO + EE/EO + EO azide-chains are linked by 4,4'bipy ligands resulting in pi-pi stacked pyridyl-columns. The azido ligand dispositions in compounds 3 and 4 make possible systems of type -AF-AF-F-, which would give rise to a topological ferromagnetic behavior. PMID- 11487313 TI - Synthesis and insertion reactions of the cyclometalated palladium-alkyl complexes Pd(CH(2)CMe(2)-o-C(6)H(4))L(2). Observation of a pentacoordinated intermediate in the insertion of SO(2). AB - Palladacycles of the type 2, have been synthesized by treatment of the corresponding chloroalkylcomplexes Pd(CH(2)CMe(2)C(6)H(5))ClL(2) (L(2) = (PMe(3))(2), 2a, or 1,5-cyclooctadiene (cod), 2b) with suitable bases, or by simple ligand exchange reactions from the cyclooctadiene derivative 2b. The metallacycles 2 react with activated alkynes and with sulfur dioxide, giving rise to different insertion products. PMID- 11487314 TI - Theoretical investigation on the electronic structure of pentacyano(L)ferrate(II) complexes with NO(+), NO, and NO(-) ligands. Redox interconversion, protonation, and cyanide-releasing reactions. AB - Reaction pathways for the one- and two-electron reductions of [Fe(CN)(5)NO](2)(-) have been investigated by means of a density functional theory (DFT) approach combined with the polarized continuum model (PCM) of solvation. In addition, UV vis spectroscopic data were obtained using ZINDO/S calculations including a point charge model simulation of solvent effects. DFT methodologies have been used to assess the thermodynamical feasibility of protonation and cyanide-release processes for the reduced species. We conclude that [Fe(CN)(5)NO](3)(-) is a stable species in aqueous solution but may release cyanide yielding [Fe(CN)(4)NO](2)(-), consistent with experimental results. On the other hand, the [Fe(CN)(5)NO](4)(-) complex turns out to be unstable in solution, yielding the product of cyanide release, [Fe(CN)(4)NO](3)(-), and/or the protonated HNO complex. All the structural and spectroscopic (IR, UV-vis) predictions for the [Fe(CN)(5)HNO](3)(-) ion are consistent with the scarce but significant experimental evidence of its presence as an intermediate in nitrogen redox interconversion chemistry. Our computed data support an Fe(II)(LS) + NO(+) assignment for [Fe(CN)(5)NO](2)(-), an Fe(II)(LS) + NO assignment for the one electron reduction product, but an Fe(I)(LS) + NO(+) for the one-electron product after dissociation of an axial cianide, and an Fe(II) + singlet NO(-) for the two electron reduction species. PMID- 11487315 TI - Preparation, characterization, and luminescence properties of gallium-metal face to-face diporphyrins (M = H(2), GaL, Ru(CO)(OH), Co). AB - The preparation and characterization of a new series of mixed metal cofacial anthracene-bridged diporphyrins (DPA) containing a GaL fragment (L = OMe, OH) and another metallic center (M = GaL, Ru(CO)(OH), Co, and H(2) (i.e. free base)) are reported. The luminescence properties at 298 and 77 K, in degassed EtOH solution, are also reported, and are characterized by a weak pipi fluorescence (2 < tau(F) < 7 ns) arising from the low energy Q-bands (S(1)-->S(0)). In the mixed diporphyrin systems, a strong pipi fluorescence is detected from the free base, while the transition metalloporphyrins of Co(II) and Ru(II) do not emit. The homobimetallic di[Ga(OMe)] species exhibits an unprecedented double (pi)(pi) fluorescence arising from the two lowest energy absorption Q-bands. On the basis of a comparison with photophysical data on GaL monoporphyrins, the weak fluorescence and absence of phosphorescence for most cases indicate efficient intramolecular quenching. To define structural features, the X-ray structures of (DPA)[Ga(OMe)](2) (2), (DPA)[Ga-(OH)-Ru(CO)] (5a), and (DPA)[Ga(OMe) Ru(MeOH)(CO)] (5b) have been obtained. The structures of 5a and 5b demonstrate an interesting aspect of the structural chemistry of these ligands related to the internal methoxide and methanolic ligands in 5b (resulting in a large interplanar separation and center-to-center distance) and the internal metal-bridging hydroxyl ligand in 5a (resulting in a small interplanar separation and center-to center distance). These data support previously reported discussions on the ability of the DPA and the DPB analogue (diporphyrinylbiphenylenyl) ligands to open and close their "bite" around the binding pocket between the porphyrin macrocycles. PMID- 11487316 TI - Novel boxlike dinuclear or chain polymeric silver(I) complexes with polypyridyl bridging ligands: syntheses, crystal structures, and spectroscopic and electrochemical properties. AB - The syntheses, characterization, crystal structures, and photophysical and electrochemical properties of two dinuclear and two polymeric Ag(I) complexes with three polypyridyl ligands, 2,3-di-2-pyridylquinoxaline (L(1)), 2,3-di-2 pyridyl-5,8-dimethoxyquinoxaline (L(2)), and 2,3,7,8-tetrakis(2 pyridyl)pyrazino[2,3-g] quinoxaline (L(3)), are described. The structures of the two boxlike dinuclear complexes with L(1) and L(2) and two chemically the same but differently crystallized one-dimensional zigzag chain coordination polymers also consisting of boxlike dinuclear subunits have been elucidated by X-ray analysis. [AgL(1)(CH(3)CN)](2)-(BF(4))(2).2CHCl(3) (1): monoclinic, C2/c; a = 28.631(2), b = 12.2259(11), c = 14.3058(12) A; beta = 99.180(2) degrees; Z = 4. [AgL(2)(CH(3)CN)(2)](2)(ClO(4))(2) (2): triclinic, P1; a = 12.3398(2), b = 13.750(2), c = 14.326(7) A; alpha = 83.494(3), beta = 74.631(3), gamma = 76.422(3) degrees; Z = 4. [[Ag(2)L(3)(NO(3))(2)].CH(3)CN](infinity) (3a): monoclinic, P2(1)/c; a = 9.5836(8), b = 13.4691(12), c = 14.0423(12) A; beta = 107.753(2) degrees; Z = 4. [Ag(2)L(3)()(NO(3))(2)](infinity) (3b): monoclinic, P2(1)/c; a = 8.4689(6), b = 16.0447(12), c = 11.7307(8) A; beta = 102.051(1) degrees; Z = 2. The structures of the dinuclear complexes 1 and 2 are similar to each other, with the two intramolecular Ag(I) centers of each complex being spanned by two ligands thus forming a unique boxlike cyclic dimer. In 1, each Ag(I) center is four-coordinated by three nitrogen atoms of two L(1) ligands and a CH(3)CN nitrogen donor, taking a distorted tetrahedral coordination geometry. The coordination environment of Ag(I) in 2 is similar to that in 1, except the formation of an additional weak coordination bond with the oxygen atom of the methoxy group of L(2). The structures of 3a,b are very similar to each other, except for the stacking patterns in the crystal lattices, and the cyclic boxlike dinuclear unit, which is similar to the structure of 1, constitutes the fundamental building block to form the one-dimensional zigzag chain structures due to the "end-on" nature of L(3). 1-3 exhibit metal-perturbed intraligand transitions in solution in 360-390 nm regions. Cyclic voltammetric studies of these complexes show the presence of reduction peak at approximately -0.5 V vs Fc(+/0). In the solid state at 77 K, they exhibit broad emission that may be assignable to originate from the metal-perturbed intraligand transitions. PMID- 11487317 TI - Synthesis, structure, and redox and catalytic properties of a new family of ruthenium complexes containing the tridentate bpea ligand. AB - We have prepared a new family of ruthenium complexes containing the bpea ligand (where bpea stands for N,N-bis(2-pyridyl)ethylamine), with general formula [Ru(bpea)(bpy)(X)](n+) (2, X = Cl(-); 3, X = H(2)O; 4, X = OH(-)), and the trisaqua complex [Ru(bpea)(H2O)(3)](2+), 6. The complexes have been characterized through elemental analyses, UV-vis and (1)H NMR spectroscopy, and electrochemical studies. For complex 3, the X-ray diffraction structure has also been solved. The compound belongs to the monoclinic P2(1)/m space group, with Z = 2, a = 7.9298(6) A, b = 18.0226(19) A, c = 10.6911(8) A, and beta = 107.549(8) degrees. The Ru metal center has a distorted octahedral geometry, with the O atom of the aquo ligand placed in a trans position with regard to the aliphatic N atom of the bpea ligand so that the molecule possesses a symmetry plane. NMR spectra show that the complex maintains its structure in aqueous solution, and that the corresponding chloro complex also has a similar structural arrangement. The pH dependence of the redox potential for the complex [Ru(bpea)(bpy)(H2O)](PF(6))(2) is reported, as well as the ability of the corresponding oxo complex to catalyze the oxidation of benzylic alcohol to benzaldehyde in both chemical and electrochemical manners. PMID- 11487318 TI - Molecular and electronic structure of octahedral o-aminophenolato and o iminobenzosemiquinonato complexes of V(V), Cr(III), Fe(III), and Co(III). Experimental determination of oxidation levels of ligands and metal ions. AB - The coordination chemistry of the ligands 2-anilino-4,6-di-tert-butylphenol, H[L(AP)], and N,N"'-bis[2-(4,6-di-tert-butylphenol]diethylenetriamine, H(2)[(L(AP))N(L(AP))], has been studied with the first-row transition metal ions V, Cr, Fe, and Co. The ligands are noninnocent in the sense that the aminophenolato parts, [L(AP)](-) and [L(AP)-H](2)(-), can be readily oxidized to their o-iminobenzosemiquinonato, [L(ISQ)](-), and o-iminobenzoquinone, [L(ISB)], forms. The following neutral octahedral complexes have been isolated as crystalline materials, and their crystal structures have been determined by X-ray crystallography at 100 K: [Cr(III)(L(ISQ))(3)] (1), [Fe(III)(L(ISQ))(3)] (2), [Co(III)(L(ISQ))(3)] (3), [V(V)(L(ISQ))(L(AP)-H)(2)] (4), [V(V)(L(AP) H)(2)(L(AP))] (5), and [V(V)O[(L(AP))N(L(AP)-H)]] (6). From variable-temperature magnetic susceptibility measurements and X-band EPR spectroscopy it has been established that they possess the ground states: 1, S = 0; 2, S = 1; 3, S = (3)/(2); 4, S = (1)/(2); 5, S = 0; 6, S = 0. The o-iminobenzosemiquinonato radicals (S(rad) = (1)/(2)) couple strongly intramolecularly antiferromagnetically to singly occupied orbitals of the t(2g) subshell at the respective metal ion but ferromagnetically to each other in 3 containing a Co(III) ion with a filled t(2g)(6) subshell. It is demonstrated that the oxidation level of the ligands and metal ions can be unequivocally determined by high-quality X-ray crystallography in conjunction with EPR, UV-vis, and Mossbauer spectroscopies. The spectro- and electrochemistry of these complexes have also been studied in detail. Metal- and ligand-based redox chemistry has been observed. The molecular and electronic structures are compared with those of their o-semiquinonato analogues. PMID- 11487319 TI - Copper(II) complexes of novel N-alkylated derivatives of cis,cis-1,3,5 triaminocyclohexane. 1. Preparation and structure. AB - Novel N,N',N' '-trialkylated derivatives of cis,cis-1,3,5-triaminocyclohexane (tach), designated tach-R(3), were prepared through alkylation of N-protected tach with subsequent acid deprotection, to afford N-methyl, N-ethyl, and N-n propyl derivatives as their trihydrobromide salts. The tach-neopentyl(3) and tach furan(3) derivatives were prepared by formation of the imine from tach and pivaldehyde or furan-2-carboxaldehyde, respectively, followed by reduction of the imine. Complexes [Cu(tach-R(3))Cl(2)] (R = Me, Et, n-Pr, CH(2)-2-thienyl, and CH(2)-2-furanyl) were prepared from CuCl(2) in MeOH or MeOH-Et(2)O solvent. Crystallographic characterization of [Cu(tach-Et(3))Br(0.8)Cl(1.2)] (Pnma, a = 8.2265(1) A, b = 12.5313(1) A, c = 15.3587(3) A, Z = 4) reveals a square-based pyramidal CuN(3)X(2) coordination sphere in which one nitrogen donor occupies the apical position at a slightly longer distance (Cu-N = 2.218(5) A) than those of the basal nitrogens (Cu-N = 2.053(2) A). The solution-phase (pH 7.4 buffered and methanol) and solid-phase structures of [Cu(tach-R(3))Cl(2)] have been studied extensively by EPR and visible-near-IR spectroscopies. The square-based pyramidal structure is retained in solution, according to correspondence of solution and solid-state data. In aqueous solution, halide is replaced by water, as indicated by the high-energy UV-vis spectral shifts and bonding parameters of [Cu(tach Et(3))](2+)(aq) derived from EPR data. The proposed aqueous-phase species, in the pH range 7.4 to 10.1, is [Cu(tach-Et(3))(H(2)O)(2)](2+). The complex [Cu(tach Me(3))](2+)(aq) does not appear to dimerize or form metal-hydroxo species at pH 7.4, in contrast to other Cu(II)-triamine complexes, e.g., [Cu(1,4,7 triazacyclononane)](2+) (aq) and [Cu(tach-H(3))](2+)(aq) (the complex of unalkylated tach). This difference is attributed to the steric effect of the N alkyl groups in the tach-R(3) series. PMID- 11487320 TI - Copper(II) complexes of novel N-alkylated derivatives of cis,cis-1,3,5 triaminocyclohexane. 2. Metal-promoted phosphate diester hydrolysis. AB - Aqueous copper(II) N,N',N' '-trimethyl-cis,cis-1,3,5-triaminocyclohexane (Cu(tach Me(3))(2+)(aq)) promotes the hydrolysis of activated phosphate diesters in aqueous medium at pH 7.2. This complex is selective for cleavage of the phosphate diester sodium bis(p-nitrophenyl) phosphate (BNPP), the rate of hydrolysis of the monoester disodium p-nitrophenyl phosphate being 1000 times slower. The observed rate acceleration of BNPP hydrolysis is slightly greater than that observed for other Cu(II) complexes, such as [Cu([9]aneN(3))Cl(2)] ([9]aneN(3) identical with 1,4,7-triazacyclononane). The rate of hydrolysis is first-order in phosphate ester at low ester concentration and second-order in [Cu(tach-Me(3))](2+)(aq), suggesting the involvement of two metal complexes in the mechanism of substrate hydrolysis. The reaction exhibits saturation kinetics with respect to BNPP concentration according to a modified Michaelis-Menten mechanism: 2CuL + S <==> LCu-S-CuL --> 2CuL + products (K(M) = 12.3 +/- 1.8 mM(2), k(cat) = (4.0 +/- 0.4) x 10(-)(4) s(-1), 50 degrees C) where CuL (triple bond) [Cu(tach-Me(3))](2+), S (triple bond) BNPP, and LCu-S-CuL is a substrate-bridged dinuclear complex. EPR data indicate that the dicopper complex is formed only in the presence of BNPP; the active LCu-S-CuL intermediate species then slowly decays to products, regenerating monomeric CuL. PMID- 11487321 TI - Kinetics and mechanism of a catalytic chloride ion effect on the dissociation of model siderophore hydroxamate-iron(III) complexes. AB - Proton-driven ligand dissociation kinetics in the presence of chloride, bromide, and nitrate ions have been investigated for model siderophore complexes of Fe(III) with the mono- and dihydroxamic acid ligands R(1)C(=O)N(OH)R(2) (R(1) = CH(3), R(2) = H; R(1) = CH(3), R(2) = CH(3); R(1) = C(6)H(5), R(2) = H; R(1) = C(6)H(5), R(2) = C(6)H(5)) and CH(3)N(OH)C(=O)[CH(2)](n)C(=O)N(OH)CH(3) (H(2)L(n); n = 2, 4, 6). Significant rate acceleration in the presence of chloride ion is observed for ligand dissociation from the bis(hydroxamate)- and mono(hydroxamate)-bound complexes. Rate acceleration was also observed in the presence of bromide and nitrate ions but to a lesser extent. A mechanism for chloride ion catalysis of ligand dissociation is proposed which involves chloride ion dependent parallel paths with transient Cl(-) coordination to Fe(III). The labilizing effect of Cl(-) results in an increase in microscopic rate constants on the order of 10(2)-10(3). Second-order rate constants for the proton driven dissociation of dinuclear Fe(III) complexes formed with H(2)L(n)() were found to vary with Fe-Fe distance. An analysis of these data permits us to propose a reactive intermediate of the structure (H(2)O)(4)Fe(L(n)())Fe(HL(n))(Cl)(OH(2))(2+) for the chloride ion dependent ligand dissociation path. Environmental and biological implications of chloride ion enhancement of Fe(III)-ligand dissociation reactions are presented. PMID- 11487322 TI - Molecular and electronic structure of [Mn(V)N(cyclam-acetato)]PF6. A combined experimental and DFT study. AB - From the reaction of Li(cyclam-acetate), MnCl(2).4H(2)O, and KPF(6) in methanol brown microcrystals of [Mn(III)Cl(cyclam-acetato)]PF(6) (1) were obtained in the presence of air (cyclam-acetic acid = 1,4,8,11-tetraazacyclotetradecane-1-acetic acid). The reaction of 1 in aqueous NH(3) solution with NaOCl produced blue crystals of [Mn(V)N(cyclam-acetato)]PF(6) (2). Complexes 1 and 2 were characterized by single-crystal X-ray crystallography, IR and Raman, electronic absorption, and (1)H, (13)C, and (15)N NMR spectroscopies. Their magnetochemistry as well as their electrochemistry have been investigated. The complexes [MnN(cyclam-acetato)](+/2+) were studied by theoretical calculations at the DFT and semiempirical levels in order to obtain more insight into the ground and excited states of the Mn(V)(triple bond)N unit. Structural and spectroscopic parameters were successfully calculated and compared to experiment. A pictorial description of the bonding has been developed. PMID- 11487323 TI - Single-molecule magnets: site-specific ligand abstraction from [Mn12O12(O2CR)16(H2O)4] and the preparation and properties of [Mn12O12(NO3)4(O2CCH2Bu(t))12(H2O)4]. AB - Site-selective carboxylate abstraction has been achieved from [Mn(12)O(12)(O(2)CR)(16)(H(2)O)(4)] complexes by treatment with HNO(3) in MeCN. The reaction of the R = Ph or CH(2)Bu(t)() complexes with 4 equiv of HNO(3) gives [Mn(12)O(12)(NO(3))(4)(O(2)CR)(12)(H(2)O)(4)] (R = CH(2)Bu(t) (6) or Ph (7)) in analytical purity. Complex 6.MeNO(2) crystallizes in monoclinic space group C2/c with the following cell parameters at -168 degrees C: a = 21.280(5), b = 34.430(8), c = 33.023(8) A, beta = 104.61(1) degrees, V = 23413 A, and Z = 8. The four NO(3)(-) groups are not disordered and are bound in bridging modes at axial positions formerly occupied by bridging carboxylate groups. (1)H NMR spectroscopy in CD(2)Cl(2) and CDCl(3) shows retention of the solid-state structure on dissolution in these solvents. DC magnetic susceptibility (chi(M)) and magnetization (M) studies have been carried out in the 2.00-300 K and 1.0-7.0 T ranges. Fits of M/Nmu(B) versus H/T plots gave S = 10, g = 1.92, and D = -0.40 cm(-1), where D is the axial zero-field splitting parameter. AC magnetic susceptibility studies on 6 have been performed in the 1.70-10.0 K range in a 3.5 Oe field oscillating at frequencies up to 1500 Hz. Out-of-phase magnetic susceptibility (chi(M)' ') signals were observed in the 4.00-8.00 K range which were frequency-dependent. Thus, 6 displays the slow magnetization relaxation diagnostic of a single-molecule magnet (SMM). The data were fit to the Arrhenius law, and this gave the effective barrier to relaxation (U(eff)) of 50.0 cm(-1) (72.0 K) and a pre-exponential (1/tau(0)) of 1.9 x 10(8) s(-1). Complex 6 also shows hysteresis in magnetization versus DC field scans, and the hysteresis loops show steps at regular intervals of magnetic field, the diagnostic evidence of field-tuned quantum tunneling of magnetization. High-frequency EPR (HFEPR) spectroscopy on oriented crystals of complex 6 shows resonances assigned to transitions between zero-field split M(s) states of the S = 10 ground state. Fitting of the data gave S = 10, g = 1.99, D = -0.46 cm(-1), and B(4)(0) = -2.0 x 10(-5), where B(4)(0) is the quartic zero-field coefficient. The combined results demonstrate that replacement of four carboxylate groups with NO(3)(-) groups leads to insignificant perturbation of the magnetic properties of the Mn(12) complex. Complex 6 should now be a useful starting point for further reactivity studies, taking advantage of the good leaving group properties of the NO(3)(-) ligands. PMID- 11487324 TI - The origin of hindered rotation around the Pt-N bond in platinum amides. AB - Several platinum amides of formula trans-[PtCl(NHAr)(PEt(3))(2)] (Ar = 3 FC(6)H(4), 2; 4-FC(6)H(4), 3; 4-ClC(6)H(4), 4; 4-IC(6)H(4), 5; 4-Cl,3-NO(2) C(6)H(3), 6) have been synthesized by reaction of [PtHCl(PEt(3))(2)] with aryl azides. All the complexes feature planar arylamido moieties and hindered rotation around the N-aryl and Pt-N bonds have been detected and separately studied. The X ray crystal structures of complexes 5 and 6 have been determined. Complex 5 crystallizes in the orthorhombic space group Pnma, with a = 23.806(4) A, b = 15.099(2) A, c = 6.7593(10) A, alpha = beta = gamma = 90 degrees, and Z = 4. Compound 6 shows an N-H...O(NO) hydrogen bond and it crystallizes in the monoclinic space group P2(1)/n, with a = 12.215(3) A, b = 8.078(2) A, c = 13.052(4) A, alpha = gamma = 90 degrees, beta = 90.057(6) degrees, and Z = 2. Except for Ar = 4-Cl,3-NO(2)-C(6)H(3), the activation energies obtained for the complexes indicate that both dynamic processes occur simultaneously with a common barrier which originates in the multiple bond character of the N-aryl bond due to a strong pi-donor behavior of the N atom in the N-aryl bond. The rotation about the Pt-N bond is unfavorable because of steric congestion with the planar amide, which can be overcome only when the aromatic ring can rotate. For the complex trans-[PtCl[NH(4-Cl,3-NO(2)-C(6)H(3))](PEt(3))(2)] the barrier to rotation is mostly due to hydrogen bond interaction between the NO(2) ortho substituent and the amide H atom. PMID- 11487325 TI - Structural and oxo-transfer reactivity differences of hexacoordinate and pentacoordinate (nitro)(tetraphenylporphinato)cobalt(III) derivatives. AB - The oxo-transfer catalyst (nitro)(pyridyl)cobalt(III) tetraphenylporphyrin has been reinvestigated by substitution of the distal pyridine ligand with 4-N,N dimethylaminopyridine and 3,5-dichloropyridine. Differences in their structures and in the reactivity of the compounds toward catalytic secondary oxo transfer were investigated by FT-IR and UV-visible spectroscopy, cyclic voltammetry, X-ray diffraction, semiempirical calculations, and reactions with alkenes in dichloromethane solution. Very modest differences in the hexacoordinate compounds' structures were predicted and observed, but the secondary oxo-transfer reactivity at the nitro ligand varies markedly with the basicity of the pyridine ligand and the position of the coordination equilibrium. Oxo transfer occurs rapidly through the pentacoordinate species (nitro)cobalt(III) tetraphenylporphyrin that is generated by dissociation of the pyridine ligand and therefore is strongly related to the Hammett parameters of these nitrogenous bases. The reactive pentacoordinate species CoTPP(NO(2)) can be generated in solution by addition of lithium perchlorate to (py)CoTPP(NO(2)) by Lewis acid base interactions or more simply by using the weaker Lewis base Cl(2)py instead of py as the distal ligand. In contrast to pentacoordinate (nitro)iron porphyrins, disproportionation reactions of CoTPP(NO(2)) compound are not evident. This pentacoordinate derivative, CoTPP(NO(2)), is reactive enough to stoichiometrically oxidize allyl bromide in minutes. Preliminary catalytic oxidation reaction studies of alkenes also indicate the involvement of both radical and nonradical oxo-transfer steps in the mechanism, suggesting formation of a peroxynitro intermediate in the reaction of the reduced CoTPP(NO) with O(2). PMID- 11487326 TI - New dinuclear catalysts Rh(2)(N-O)(2)[(C6H4)P(C6H5)2]2 with imidate ligands: synthesis and isomerization from head-to-tail to head-to-head configuration of the imidate ligands. AB - Two new dirhodium(II) catalysts of general formula Rh(2)(N O)(2)[(C(6)H(4))P(C(6)H(5))(2)](2) (N-O = C(4)H(4)NO(2)) are prepared, starting from Rh(2)(O(2)CCH(3))(2)(PC)(2)L(2) [PC = (C(6)H(4))P(C(6)H(5))(2) (head-to-tail arrangement); L = HO(2)CCH(3)]. The thermal reaction of Rh(2)(O(2)CCH(3))(2)(PC)(2).L(2) with the neutral succinimide stereoselectively gives one compound that according to the X-ray structure determination has the formula Rh(2)(C(4)H(4)NO(2))(2)[(C(6)H(4))P(C(6)H(5))(2)](2) (1). It corresponds to the polar isomer with two bridging imidate ligands in a head-to-head configuration. However, stepwise reaction of Rh(2)(O(2)CCH(3))(2)(PC)(2).L(2) with (CH(3))(3)SiCl and potassium succinimidate yields a mixture of 1 and one of the two possible isomers (structure B) with a head-to-tail configuration of the imidate ligands, Rh(2)(C(4)H(4)NO(2))(2)[(C(6)H(4))P(C(6)H(5))(2)](2) (2), also characterized by X-ray methods. In solution, compound 2 undergoes slow isomerization to 1; the rate of this process is enhanced by the presence of acetonitrile. Compounds 1 and 2 are obtained as pure enantiomers starting from (M)- and (P)-Rh(2)(O(2)CCH(3))(2)(PC)(2).L(2) rather than from the racemic mixture. Their enantioselectivities in cyclopropanation of 1-diazo-5-penten-2-one are similar to those reported for the dirhodium amidate catalysts. PMID- 11487327 TI - Interactions of metal ions with water: ab initio molecular orbital studies of structure, vibrational frequencies, charge distributions, bonding enthalpies, and deprotonation enthalpies. 2. Monohydroxides. AB - The formation and properties of a wide range of metal ion monohydroxides, M(n)(+)[OH(-)], where n = 1 and 2, have been studied by ab initio molecular orbital calculations at the MP2(FULL)/6-311++G**//MP2(FULL)/6-311++G** and CCSD(T)(FULL)/6-311++G**//MP2(FULL)/6-311++G** computational levels. The ions M(n)()(+) are from groups 1A, 2A, 3A, and 4A in the second, third, and fourth periods of the Periodic Table and from the first transition series. Geometrical parameters, vibrational frequencies, atomic charge distributions, orbital occupancies, and bonding enthalpies are reported. The M(n)(+)-O distances are shorter in the hydroxides than in the corresponding hydrates (published previously as Part 1, Inorg. Chem. 1998, 37, 4421-4431) due to a greater electrostatic interaction in the hydroxides. The natural bond orbitals for most of the first-row transition metal ion hydroxides do not contain a formal metal oxygen bonding orbital; nevertheless the atomic charge distributions show that for both n = 1 and 2 a significant amount of electron density is consistently transferred from the hydroxide ion to the bound metal ion. Deprotonation enthalpies for the hydrates have been evaluated according to the simple dissociation process, M(n)(+)[OH(2)] --> M(n)(+)[OH(-)] + H(+), and also via proton transfer to another water molecule, M(n)(+)[OH(2)] + H(2)O --> M(n)(+)[OH( )] + H(3)O(+). The drastic reduction in these deprotonation enthalpies as H(2)O molecules are sequentially bonded in the first coordination shell of the metal ion (amounting to 71, 64, 85, and 91 kcal/mol for the bonding of six water molecules to Mg(2+), Ca(2+), Mn(2+), and Zn(2+), respectively) is found to be due to the greater decrease in the bonding enthalpies for the hydroxides relative to the hydrates. Proton transfer to bases other than water, for example side chain groups of certain amino acids, could more than offset the decrease in deprotonation energy due to the filling of the first coordination shell. Linear relationships have been found between the pK(a) values for ionization of the Mg(2+), Ca(2+), Mn(2+), Fe(2+), Co(2+), Ni(2+), Cu(2+), and Zn(2+) aquo ions, and Delta for the bonding of the first water molecule, for the bonding of the hydroxide ion, and for proton dissociation from the monohydrate. Similar relationships have also been found between the pK(a) values and the reciprocal of the M-O bond lengths in both the monohydrates and hydroxides. Thus the ionization of metal hydrates in water echoes the properties of the monomeric species M(n)(+)[OH(2)]. PMID- 11487328 TI - Heterobimetallic oxalato-bridged Cu(II)Re(IV) complexes. Synthesis, crystal structure and magnetic properties. AB - Three copper(II)-rhenium(IV) bimetallic complexes of formula [ReCl(4)(mu ox)Cu(phen)(2)] (1), [ReCl(4)(mu-ox)Cu(phen)(2)].CH(3)CN (2), and [ReCl(4)(mu ox)Cu(terpy) (H(2)O)][ReCl(4)(mu-ox)Cu(terpy)(CH(3)CN)] (3) (ox = oxalate anion, phen = 1,10-phenanthroline, and terpy = 2,2':6,2"- terpyridine) have been synthesized and their crystal structures determined by single-crystal X-ray diffraction. Complex 1 crystallizes in the triclinic system, space group P(-1), with a = 9.776(2), b = 11.744(3), c = 14.183(3) A, alpha =102.09(2) degrees, beta = 109.42(2) degrees, gamma = 107.11(2) degrees, and Z = 2, whereas 2 and 3 crystallize in the monoclinic system, space groups P2(1)/n and P2(1)/c, respectively, with a = 12.837(3), b = 17.761(4), c = 12.914(3) A, beta = 91.32(2) degrees, and Z = 4 for 2, and a = 8.930(2), b = 18.543(4), c = 27.503(6) A, beta = 94.67(2) degrees, and Z = 4 for 3. The structures of 1 and 2 are made up of neutral [ReCl(4)(mu-ox)Cu(phen)(2)] bimetallic units. Re(IV) and Cu(II) metal ions exhibit distorted octahedral coordination geometries, being bridged by a bis(bidentate) oxalato ligand. The presence of acetonitrile molecules of crystallization in 2 causes a somewhat greater separation between the bimetallic complexes and a different packing of these units in the crystal structure with respect to 1. The copper-rhenium separation across oxalato is 5.628(2) in 1 and 5.649(3) A in 2. The structure of 3 is made up of two different and neutral bimetallic units, [ReCl(4)(mu-ox)Cu(terpy)(H(2)O)] and [ReCl(4)(mu ox)Cu(terpy)(CH(3)CN)]. In the first one, the oxalate group behaves as a bis(bidentate) ligand occupying one equatorial and one axial position in the elongated octahedral environment of Cu(II). The water molecule is axially coordinated. In the second one, the oxalate group behaves as a bidentate/monodentate ligand occupying the axial position in the square pyramidal environment of Cu(II). The acetonitrile molecule occupies a basal coordination position around the copper atom. These units are arranged in such a way that a chlorine atom of the first unit (Cl(1)) points toward the copper atom (Cu(2))of the second one (3.077(2) A for Cl(1)(.)Cu(2)), forming a tetranuclear species. The copper-rhenium separation across bis(didentate) oxalato is 5.504(3) A, whereas that through bidentate/monodentate oxalato is 5.436(2) A. The magnetic behavior of 2 and 3 has been investigated over the temperature range 1.8-300 K. A very weak and nearly identical antiferromagnetic coupling between Re(IV) and Cu(II) through bis(bidentate) oxalato occurs in 2 (J = -0.90 cm(-1)) and 3 (J = 0.83 cm(-1)); it is ferromagnetic in 3 through both the bidentate-monodentate oxalato (J = +5.60 cm(-1)) and the chloro (J = +0.70 cm(-1)) bridges. PMID- 11487329 TI - Synthesis, characterization, and stereochemistry of oxorhenium(V) complexes with 2-aminoethanethiolate. AB - A series of oxorhenium(V) complexes with 2-aminoethanethiolate (aet), [ReO(aet N,S)(D-pen-N,O,S)] (2), [[ReO(aet-N,S)(2)](2)O] (3), [ReO(Cl)(aet-N,S)(2)] (4), and [ReO(aet-N,S)(Haet-S)(2)]Cl(2) ([5]Cl(2)) was newly prepared starting from ReO(4)(-). The reaction of NH(4)ReO(4) with a 1:1 mixture of Haet.HCl and D H(2)pen (D-penicillamine) in the presence of SnCl(2).2H(2)O in water gave 2, 3, and the known complex [ReO(D-Hpen-N,S)(D-pen-N,O,S)] (1). These complexes were fractionally precipitated by controlling the pH of the reaction solution. The complex 2 was also prepared in a higher yield by a similar reaction using methanol as a solvent. The crystal structure of 2 was determined by X-ray crystallography; 2 crystallizes in the tetragonal space group P4(3) with a = 9.621(1), c = 12.911(1) A, V = 1195.0(3) A(3), and Z = 4. The oxorhenium(V) core in 2 is coordinated by a bidentate-N,S aet ligand and a tridentate-N,O,S D-pen ligand, having a distorted octahedral geometry with a cis-N cis-S configuration in the equatorial plane perpendicular to the O-Re-O axis. The 1:2 reaction of NH(4)ReO(4) with Haet.HCl in the presence of SnCl(2).2H(2)O in methanol produced 4, which is interconvertible with 3, while the corresponding 1:3 reaction resulted in the isolation of [5]Cl(2). The complexes 4 and 5 were also structurally characterized; 4 crystallizes in the monoclinic space group P2(1)/c with a = 6.839(1), b = 10.0704(6), c = 14.1075(8) A, beta = 91.729(8) degrees, V = 971.2(2) A(3), and Z = 4, while [5]Cl(2) crystallizes in the triclinic space group P1 with a = 11.938(3), b = 12.366(3), c = 5.819(1) A, alpha = 102.71(2), beta = 101.28(2), gamma = 75.41(2) degrees, V = 802.0(3) A(3), and Z = 2. In 4, the oxorhenium(V) core is octahedrally coordinated by two bidentate-N,S aet ligands, which form a cis-N cis-S configurational equatorial plane with a Cl(-) ion trans to the oxo ligand. On the other hand, the oxorhenium(V) core in [5](2+) is coordinated by one bidenate-N,S aet and two monodentate-S Haet ligands, having a distorted trigonal-bipyramidal geometry with S and N donors at the apical positions. PMID- 11487330 TI - Nickel dithiolenes revisited: structures and electron distribution from density functional theory for the three-member electron-transfer series [Ni(S2C2Me2)2]0,1 ,2-. AB - The complexes [Ni(S2C2Me2)2](z) (z = 0, 1-, 2-) have been isolated for the purpose of investigating their electronic structures in a reversible three-member electron-transfer series. Members are interrelated by reversible redox reactions with E(1/2)(0/1-) = -0.15 V and E(1/2)(1-/2-) = -1.05 V versus SCE in acetonitrile. The three complexes have nearly planar structures of idealized D(2)(h) symmetry. As the series is traversed in the reducing direction, Ni-S and C-S bond lengths increase; the chelate ring C-C bond length decreases from the neutral complex to the monoanion and does not change significantly in the dianion. Structural trends are compared with previous results for [Ni(S2C2R2)2)](1-,2-). Following the geometrical changes, values of nu(Ni)(-)(S) and nu(C)(-)(S) decrease, while the value of nu(C)(-)(C) increases with increased reduction. Geometry optimizations at the density functional theory (DFT) level were performed for all members of the series. Geometrical parameters obtained from the calculations are in good agreement with the experimental findings. The 5b(2g) orbital was identified as the LUMO in [Ni(S2C2Me2)2], the SOMO in [Ni(S2C2Me2)2](1-), and the HOMO in [Ni(S2C2Me2)2]2-. Unlike in the situation in the [M(CO)2-(S2C2Me2)2]z series (M = Mo, W; z = 0, 1-, 2-), the apparent contribution from the metal d orbital in the electroactive orbital is not constant. In the present series, the d(xz) contribution increases from 13 to 20 to 39% upon passing from the neutral to the monoanionic to the dianionic complex. Accurate calculation of EPR g-values of [Ni(S2C2Me2)2]1- by DFT serves as a test for the reliability of the electronic structure calculations. PMID- 11487331 TI - Coordination chemistry of amine bis(phenolate) titanium complexes: tuning complex type and structure by ligand modification. AB - The coordination chemistry of titanium(IV) complexes of amine bis(phenolate) ligands was investigated by synthesizing various types of complexes and analyzing them specroscopically and structurally. Steric effects of tridentate [ONO]- and tetradentate [ONNO]-type ligands were studied by reacting the ligand precursors with titanium tetra(isopropoxide). [ONNO]-type ligands featuring an amine donor located on a pendant arm led to octahedral bis(isopropoxide) complexes, regardless of the steric bulk around the metal. Several such complexes having varying steric crowding were thus synthesized. On the other hand, steric effects were found to play a major role in determining the complex constitution when [ONO]-type ligands, featuring no side donor, were involved. Relatively sterically undemanding ligands led to octahedral bis(homoleptic) complexes, whereas increased steric bulk resulted in the formation of pentacoordinate bis(isopropoxide) complexes. These pentacoordinate complexes readily lead to bis(heteroleptic) complexes by reaction with nonsterically demanding [ONO]- and [ONNO]-type ligand precursors. In the latter case the sidearm nitrogen remains uncoordinated to the metal. The bis(isopropoxide) complexes of the [ONNO]-type ligands may also lead to bis(heteroleptic) complexes, however, these reactions are much slower. PMID- 11487332 TI - Synthesis, structure, and reactions of binuclear gold(I) complexes containing two different bridging ligands. AB - The binuclear cycloaurated compounds [Au(2)(mu-C(6)H(3)-2-PPh(2)-n-Me)(2)] (n = 5, 1a; n = 6, 1b) react with the digold(I) complexes [Au(2)(mu S(2)CN(n)()Bu(2))(2)] and [Au(2)(mu-dppm)(2)](PF(6))(2) to give heterobridged dinuclear complexes [Au(2)(mu-C(6)H(3)-2-PPh(2)-n-Me)(mu-S(2)CN(n)Bu(2))] (n = 5, 5a; n = 6, 5b) and [Au(2)(mu-C(6)H(3)-2-PPh(2)-n-Me)(mu-dppm)]PF(6), (n = 5, 9a; n = 6, 9b), respectively. Complex 5a exists in the solid state as an infinite zigzag chain of dimeric units with intramolecular Au-Au separations of 2.8331(3) and 2.8243(3) A for independent molecules and intermolecular Au-Au separations of 3.0653(3) and 3.1304(3) A. Both 5a and 5b undergo oxidative addition with halogens to give the heterovalent, gold(I)-gold(III) compounds [XAu(I)(mu-2 Ph(2)PC(6)H(3)-n-Me)Au(III)X(eta(2)-S(2)CN(n)Bu(2))] [n = 5, X = Cl (6a), I (8a); n = 6, X = Cl (6b), Br (7b), I (8b)]. Compound 8a has been shown by X-ray crystallography to contain a gold(III) atom coordinated in a planar array by bidentate, chelating di-n-butyldithiocarbamate, iodide, and the sigma-aryl carbon atom, together with a gold(I) atom that is linearly coordinated by the phosphorus atom of the arylphosphine and by iodide. The intramolecular gold-gold distance of 3.2201(3) A indicates little or no interaction between the metal atoms. In contrast to the behavior of the homobridged complexes 1a and 1b, the heterobridged dithiocarbamate complexes 5a and 5b give structurally similar products on reaction with halogens, irrespective of the position of the ring methyl substituent. Crystal data for [Au(2)(mu-C(6)H(3)-2-PPh(2)-5-Me)(mu S(2)CN(n)Bu(2))] (5a): triclinic, space group P1 (No. 2), with a = 11.3398(1), b = 15.9750(2), c = 16.4400(3) A, alpha = 91.0735(9), beta = 109.3130(7), gamma = 90.7666(8) degrees, V = 2809.47(6) A(3), and Z = 4. Crystal data for [IAu(I)(mu-2 Ph(2)PC(6)H(3)-5-Me)Au(III)I(eta(2)- S(2)CN(n)Bu(2))] (8a): triclinic, space group P1 (No. 2), with a = 8.6136(2), b = 9.3273, c = 21.1518(4) A, alpha = 84.008(1), beta = 84.945(1), gamma = 75.181(1) degrees, V = 1630.54(6) A(3), and Z = 2. PMID- 11487333 TI - Structural diversity in a family of copper(I) thioether complexes. AB - Copper(I) complexes of the tridentate thioether ligands [PhB(CH(2)SCH(3))(3)] (abbreviated PhTt), [PhB(CH(2)SPh)(3)] (PhTt(Ph)), [PhB(CH(2)S(t)()Bu)(3)] (PhTt(t)()(Bu)), and [PhB(CH(2)S(p)()Tol)(3)] (PhTt(p)()(Tol)) and bidentate thioether ligands [Ph(2)B(CH(2)SCH(3))(2)] (Ph(2)Bt), [Et(2)B(CH(2)SCH(3))(2)] (Et(2)Bt), and [Ph(2)B(CH(2)SPh)(2)] (Ph(2)Bt(Ph)) have been prepared and characterized. The solution and solid state structures are highly sensitive to the identity of the borato ligand employed. Ligands possessing the smaller (methylthio)methyl donors, [PhTt] and [Ph(2)Bt], yielded tetrameric species, [(PhTt)Cu](4) and [(Ph(2)Bt)Cu](4), containing both terminal and bridging thioether ligation. The ligands containing the larger (arylthio)methyl groups, [PhTt(Ph)] and [PhTt(p)()(Tol)], form monomeric [PhTt(Ar)]Cu(NCCH(3)) in solution and one-dimensional extended structures in the solid state. Each complex type reacted cleanly with acetonitrile, pyridine, or triphenylphosphine generating the corresponding four-coordinate monomer, of which [PhTt(Ph)]Cu(PPh(3)), [PhTt(p)()(Tol)]Cu(PPh(3)), and [Et(2)Bt]Cu(PPh(3))(2) have been structurally characterized. PMID- 11487334 TI - 1H and (17)O NMR detection of a lanthanide-bound water molecule at ambient temperatures in pure water as solvent. AB - Lanthanide complexes of a tetra-amide derivative of DOTA (structure 4 in text) with four extended carboxymethyl esters have been characterized by X-ray crystallography and multinuclear NMR spectroscopy. [Eu(4)(H(2)O)](triflate)(3) crystallized from water in the monoclinic, P(21/)(c) space group (a = 10.366 A, b = 22.504 A, c = 23.975 A, and beta = 97.05 degrees ). The Eu(3+) cation is bound to four macrocyclic nitrogen atoms (mean Eu-N = 2.627 A) and four amide oxygen atoms (mean Eu-O(amide) = 2.335 A) in a square antiprismatic geometry with a twist angle of 38.5 degrees between the N4 and O4 planes. A single bound water molecule (Eu-O(W) = 2.414 A) occupies a typical monocapped position on the O4 surface. In pure water, resonances corresponding to a single Eu(3+)-bound water molecule were observed in the (1)H (53 ppm) and (17)O (-897 ppm) NMR spectra of [Eu(4)(H(2)O)](triflate)(3) at 25 degrees C. A fit of the temperature-dependent Eu(3+)-bound (1)H and (17)O water resonance line widths in acetonitrile-d(3) (containing 4% v/v (17)O enriched water) gave identical lifetimes (tau(m)(298)) of 789 +/- 50 micros (in water as solvent; a line shape analysis of the Eu(3+) bound water resonance gave a tau(m)(298) = 382 +/- 5 micros). Slow water exchange was also evidenced by the water proton relaxivity of Gd(4) (R(1) = 2.2 mM(-1) s( 1), a value characteristic of pure outer-sphere relaxation at 25 degrees C). With increasing temperature, the inner-sphere contribution gradually increased due to accelerated chemical exchange between bound water and bulk water protons. A fitting of the relaxation data (T(1)) to standard SBM theory gave a water proton lifetime (tau(m)(298)) of 159 micros, somewhat shorter than the value determined by high-resolution (1)H and (17)O NMR of Eu(4). Exchange of the bound water protons in Gd(4) with bulk water protons was catalyzed by addition of exogenous phosphate at 25 degrees C (R(1) increased to 10.0 mM(-1) s(-1) in the presence of 1500-fold excess HPO(4)(2-)). PMID- 11487335 TI - Conformational isomerism and weak molecular and magnetic interactions in ternary copper(II) complexes of [Cu(AA)L']ClO4.nH2O, where AA = L-phenylalanine and L histidine, L' = 1,10-phenanthroline and 2,2-bipyridine, and n = 1 or 1.5: synthesis, single-crystal X-ray structures, and magnetic resonance investigations. AB - Weak molecular and magnetic exchange interactions in ternary copper(II) complexes, viz., [Cu(L-phe)(phen)(H(2)O)]ClO(4) (1), [Cu(L phe)(bpy)(H(2)O)]ClO(4) (2), and [Cu(L-his)(bpy)]ClO(4).1.5H(2)O (3), where L-phe = L-phenylalanine, L-his = L-histidine, phen = 1,10-phenanthroline, and bpy = 2,2'-bipyridine, have been investigated. Single-crystal X-ray structures reveal that complex 2 crystallizes in a monoclinic space group P2(1), with unit cell parameters a = 7.422(7) A, b = 11.397(5) A, c = 12.610(2) A, beta = 102.10(5) degrees, V = 1043.0(11) A(3), Z = 2, R = 0.0574, and R(w) = 0.1657. Complex 3 crystallizes in a monoclinic space group C2, with a = 18.834(6) A, b = 10.563(4) A, c = 11.039(3) A, beta = 115.23(2) degrees, V = 1986.6(11) A(3), Z = 4, R = 0.0466, and R(w) = 0.1211. Molecules of 2, in the solid state, are self-assembled via weak intra- and intermolecular pi-pi stacking and H-bonding interactions. Molecules of 3 exhibit intermolecular dimeric association with the Cu.Cu separation being 3.811 A. X-ray structures and (1)H NMR studies reveal conformational isomerism in both solid and liquid states of complexes 1 and 2. The aromatic side chain of L-phe in 1 and 2 adopts either a "folded" (A) or an "extended" (B) conformation. Variable-temperature (1)H NMR and spin lattice relaxation measurements point out interconversion between conformations A and B at temperatures above 323 K. The change in molecular conformation induces a change in the electron density at the site of copper and band gap energy between HOMO and LUMO orbitals. Interestingly, in spite of paramagnetic nature, complexes 1 and 2 are amenable for both EPR and (1)H NMR spectroscopic studies. Single crystal EPR spectra of 2 in three orthogonal planes are consistent with three dimensional magnetic behavior. Intramolecular exchange dominates the dipolar interactions. The EPR spectra of 3 correspond to weak magnetic interactions between associated dimeric units. The structural and magnetic resonance investigations together reveal that the weak pi-pi stacking interactions are the electronic pathways for magnetic interactions in 1-3. PMID- 11487336 TI - IR and EXAFS spectroscopic studies of glyphosate protonation and copper(II) complexes of glyphosate in aqueous solution. AB - The varying degrees of protonation of N-(phosphonomethyl)glycine (PMG, glyphosate) were investigated with infrared (IR) spectroscopy and ab initio frequency calculations. The zwitterionic nature of PMG in solution was confirmed, and intramolecular hydrogen bonding was identified. Successive protonation of the PMG molecule follows the order amine, phosphonate, carboxylate. Intramolecular hydrogen bonding is indicated to exist at all stages of protonation: between both RCO(2-) and RNH(2)(+) and RPO(3)(2-) and RNH(2+) in HL(2)(-) (where L represents the ligand PMG); between RCO(2)(-) and RNH(2)(+) in H(2)L(-); predominantly between RPO(3)(2-) and RNH(2)(+) in H(3)L. There are strong indications that the zwitterion is intact throughout the pH range investigated. Results from IR and extended X-ray absorption fine structure (EXAFS) spectroscopies provide new evidence for structures of N-(phosphonomethyl)glycinecopper(II) complexes. The structures of 1:1 complexes, CuL(-) and CuHL, are essentially the same, differing only in protonation of the phosphonate group. Copper(II) lies at the center of a Jahn-Teller distorted octahedron with all three donor groups (amine, carboxylate, phosphonate) of PMG chelating with copper(II) to form two five-membered chelate rings oriented in the equatorial plane. EXAFS indicates that oxygen (most likely a water molecule) is a fourth ligand, which would thus occupy the fourth corner in the equatorial plane of the elongated octahedron. CuL(2)(4-) most probably forms an isomeric mixture in solution, and there are indications that this mixture is dominated by complexes where two PMG ligands are bound to copper(II) via equatorial and axial positions, with both phosphonate and carboxylate donor groups responsible for chelation at axial positions. PMID- 11487337 TI - Chelators for radioimmunotherapy: I. NMR and ab initio calculation studies on 1,4,7,10-tetra(carboxyethyl)-1,4,7,10-tetraazacyclododecane (DO4Pr) and 1,4,7 tris(carboxymethyl)-10-(carboxyethyl)-1,4,7,10-tetraazacyclododecane (DO3A1Pr). AB - This work describes the modification of the chelating agent 1,4,7,10 tetraazacyclododecane-N,N',N' ',N' "-tetraacetic acid (DOTA) to improve the rate of metal loading for radioimmunotherapy applications. Previous ab initio calculations predicted that the compounds 1,4,7,10-tetra(carboxyethyl)-1,4,7,10 tetraazacyclododecane (DO4Pr) and 1,4,7-tris(carboxymethyl)-10-(carboxyethyl) 1,4,7,10-tetraazacyclododecane (DO3A1Pr) have a ca. 2000-fold improvement in yttrium metal loading rates compared to those of DOTA (Jang, Y. H.; Blanco, M.; Dasgupta, S.; Keire, D. A.; Shively, J. E.; Goddard, W. A., III. J. Am. Chem. Soc. 1999, 121, 6142-6151). In this study, we report the synthesis, purification, (1)H-NMR chemical shift assignments, pK(a) values, metal loading rate measurements, and additional ab initio calculations of these two compounds. The yttrium loading rates of DO3A1Pr are approximately twice those of DOTA, at pH 4.6 and 37 degrees C. The NMR data indicates that the DO4Pr analogue forms a stable type I complex but does not form a type II complex. The new ab initio calculations performed on DO4Pr and DO3A1Pr indicate that the rate-determining step is the deprotonation of the first macrocycle amine proton, not the second proton as assumed in the previous calculations. The new calculations predict an improvement in the rate of metal loading that more closely matches the experimentally observed change in the rate. PMID- 11487338 TI - Ferromagnetic exchange in the ground and (4)A(1) excited states of (Et(4)N)(3)Fe(2)F(9). A magnetic and optical spectroscopic study. AB - The synthesis, crystal growth, magnetic susceptibility, and polarized optical absorption spectra in the visible and near UV of (Et(4)N)(3)Fe(2)F(9) are reported. From single-crystal magnetic susceptibility data and high-resolution absorption spectra in the region of the (6)A(1) --> (4)A(1) spin-flip transition, exchange splittings in the ground and excited states are derived. Ferromagnetic ground and excited state exchange parameters J(GS) = -1.55 cm(-1) and J(ES) = 0.53 cm(-1) are determined, respectively, and the relevant orbital contributions to the net exchange are derived from the spectra. The results are compared with those reported earlier for the structurally and electronically analogous [Mn(2)X(9)](5-) pairs in CsMgX(3):Mn(2+) (X = Cl(-), Br(-)), in which the splittings are antiferromagnetic. This major difference is found to be due to the increased metal charge of Fe(3+) compared to Mn(2+), leading to orbital contraction and thus to a strong decrease of the orbital overlaps and hence the antiferromagnetic interactions. PMID- 11487339 TI - Rhodium-catalyzed dehydrocoupling of the sterically encumbered phosphine-borane adduct tBu(2)PH.BH(3): synthesis of the linear dimers tBu(2)PH-BH(2)-tBu(2)P BH(3) and tBu(2)PH-BH(2)-tBu(2)P-BH(2)Cl. AB - The dehydrocoupling of the sterically hindered phosphine-borane adduct tBu(2)PH.BH(3) above 140 degrees C is catalyzed by the rhodium complexes [Rh(1,5 cod)(2)][OTf] or Rh(6)(CO)(16) to give the four-membered chain tBu(2)PH-BH(2) tBu(2)P-BH(3) (1), which was isolated in 60% yield and characterized by multinuclear NMR spectroscopy, mass spectrometry, and elemental analysis. Thermolysis of 1 in the temperature range 175-180 degrees C led to partial decomposition and the formation of tBu(2)PH.BH(3). When the dehydrocoupling of tBu(2)PH.BH(3) was performed in the presence of [[Rh(mu-Cl)(1,5-cod)](2)] or RhCl(3) hydrate, the chlorinated compound tBu(2)PH-BH(2)-tBu(2)P-BH(2)Cl (2) was formed which could not be obtained free of 1. The molecular structures of tBu(2)PH.BH(3), tBu(2)PH-BH(2)-tBu(2)P-BH(3) (1), and tBu(2)PH-BH(2)-tBu(2)P BH(2)Cl (2) together with 1 were determined by single-crystal X-ray diffraction studies. PMID- 11487340 TI - Cooperative processes governing formation of small pentanuclear lanthanide(III) nanoclusters and energy transport within and between them. AB - Syntheses, lanthanide quantitative analyses, mass spectrometry and luminescence spectroscopy, and decay dynamics of crystals containing pentanuclear hetero lanthanide(III) nanoclusters [(Ln'(5-x)Ln(x))(NO(3))(6)(mu(5)-OH)(mu(4)-L)(2)] (0 < or = x < or = 5), Ln' = Eu or Tb; Ln = La-Nd, Sm-Ho (hereafter Ln'(5-x) Ln(x)) were undertaken in search of information on factors governing self-assembly processes by which the clusters are formed and electronic interactions within and between them. The data obtained are consistent with the self-assembly of Ln'(5-x) Ln(x) nanoclusters being a concerted process featuring a profound expression of complementarity among mutually bridging [Ln(mu(4)-L](-) and [Ln(NO(3))(2)](+) components. The energy transport regime in crystals of Eu(5-x) Ln(x) is in the dynamic regime when x = 0 or Ln = La and, at 293 K, Ln = Dy, despite the presence of two crystallographically different Eu(3+) coordination environments which give rise to a doublet in the excitation and emission spectra of Eu(3+)((5)D(0)). The luminescence decay behavior of Eu(3+)((5)D(0)) in Eu(5-x) Ln(x) (Ln = Dy (for 77 K), Sm) is intermediate between the static and dynamic limits and reveals extensive electronic coupling among lanthanide ions, including many-body processes at relatively high Dy(3+) or Sm(3+) concentrations. PMID- 11487341 TI - Crystal chemistry of the 4,4'-dimethyl-2,2'bipyridine/copper bromide system. AB - The reaction of 4,4'-dimethyl-2,2-bipyridine (henceforth dmbp) with copper(I) and/or copper(II) bromide under a wide variety of conditions has led to the isolation of 10 different crystalline materials. These include one Cu(I) salt, [Cu(dmbp)(2)]Br (a distorted tetrahedral Cu species and a lattice Br(-) ion); two mixed valence Cu(I,II) compounds, [Cu(dmbp)(2)Br][CuBr(2)] (discrete 5 coordinated Cu(II) and linear Cu(I) species) and Cu(dmbp)(2)BrCu(2)Br(3) (linked 5-coordinate Cu(II) and trigonal planar Cu(I) species); and seven Cu(II) compounds, (dmbp)CuBr(2) (stacked planar monomers), [(dmbp)CuBr(2)](2)(five coordinate bibridged dimers), (dmbp)Cu(2)Br(4) (stacked planar bibridged dimers), (dmbp)CuBr(2)(DMSO) (five coordinate monomers), [Cu(dmbp)(2)Br]OH.5(1)/(2)H(2)O and [Cu(dmbp)(2)Br](Br/OH).5(1)/(2)H(2)O (five coordinate monomers), and (dmbpH(2))CuBr(4).H(2)O (distorted tetrahedral monomers). The crystal structure determinations of these materials are reported. A common thread in their structural chemistry is the supramolecular architecture developed through interdigitation of the dmbp rings on neighboring molecular species. The interdigitation leads to layer structures in many of the materials. The distances between the interdigitated dmbp rings are in the range 3.4-3.7 A. The Cu(dmbp)(2)Br(+) species exhibits an exceptionally large distortion from tetrahedral geometry due to deviation of the dihedral angle between the mean planes of the Cu(dmbp) fragments from 90 degrees. The Cu(dmbp)(2)Br(+) cations have distorted trigonal bipyramidal geometry, the Br(-) ion occupying an equatorial position. The length of the Cu-Br bond in the Cu(dmbp)(2)Br(+) species is correlated with the change in dihedral angle between the planes of the two dmbp ligands. The mono-dmbp complexes show a greater variation in coordination geometry for the Cu(II) species, including distorted trigonal bipyramidal and augmented square planar 4 + 1 and 4 + 2 coordination. PMID- 11487342 TI - Syntheses and structural characterizations of sheet- and column-like lanthanide transition metal arrays: the effect of hydrogen bonding on the structure when K(+) is replaced by [NH4]+. AB - Sheet- and column-like cyanide bridged lanthanide-transition metal arrays were synthesized through metathesis reactions between anhydrous LnCl(3) (Ln = Eu, Yb) and A(2)[M(CN)(4)] (A = K(+), NH(4)(+); M = Ni, Pt) in a 1:2 molar ratio in DMF (DMF = N,N-dimethylformamide) solution. Single-crystal X-ray analysis revealed that complexes of formula [K(DMF)(7)Ln[M(CN)(4)](2)](infinity) (Ln = Eu, M = Ni, 1; Ln = Yb, M = Pt, 2) consist of infinite layers of neutral, puckered sheets that contain hexagonal rings of composition [(DMF)(10)Ln(2)[M(CN)(4)](3)](6) with interstitial (DMF)(4)K(2)[M(CN)(4)] units located between the layers. The sheet structure is generated through the repeating (DMF)(10)Ln(2)[M(CN)(4)](3) unit with trans cyanide ligands in [M(CN)(4)](2)(-) serving as bridges. The column like complex [(NH(4))(DMF)(4)Yb[Pt(CN)(4)](2)](infinity), 3, is formed when NH(4)(+) replaces K(+). It consists of infinite, negatively charged, square, parallel columns bundled through N-H...NC hydrogen bonds between NH(4)(+) and terminal CN from the columns. Cis cyanide ligands in [Pt(CN)(4)](2)(-) units serve as bridges. Complex 3 is the first known example where Ln(III) centers are coordinated to four [M(CN)(4)](2)(-) units. Bicapped (square face) trigonal prismatic coordination geometries were observed for Ln(III) centers in 1 and 2. Square antiprismatic geometry for Yb(III) centers are observed in 3. Crystal data for 1: triclinic space group P1, a = 8.797(2) A, b = 15.621(3) A, c = 17.973(6) A, alpha = 105.48(2) degrees, beta = 98.60(2) degrees, gamma = 98.15(2) degrees, Z = 2. Crystal data for 2: triclinic space group P1, a = 8.825(1) A, b = 15.673(1) A, c = 17.946(1) A, alpha = 105.46(2) degrees, beta = 99.10(1) degrees, gamma = 98.59(1) degrees, Z = 2. Crystal data for 3: monoclinic space group P2(1)/c, a = 9.032(1) A, b = 29.062(1) A, c = 15.316(1) A, beta = 94.51(1) degrees, Z = 2. PMID- 11487343 TI - Linkage and redox isomerism in ruthenium complexes of catecholate, semi-quinone, and o-acylphenolate ligands derived from tri- and tetrahydroxy-9,10 anthracenediones. AB - The complexes Ru(CO)(2)L(2)(PHAQ-2H) (PHAQ = 1,2,4-trihydroxy-9,10 anthracenedione (PUR), 1,2,3- trihydroxy-9,10-anthracenedione (AG), and 1,2,5,8 tetrahydroxy-9,10-anthracenedione (QAL); L = PPh(3), PCy(3), PBu(3)), and Ru(CO)(dppe)(PBu(3))(PHAQ-2H), containing catecholate-type ligands were prepared. The complex Ru(CO)(2)(PBu(3))(2)(AG-2H) crystallizes in the space group P2(1)/n (No. 14 var) with a = 13.317(2), b = 15.628(2), c = 21.076(3) A, beta = 101.660(10) degrees, Z = 4; the crystal structure shows it to contain a 2,3 catecholate ligand. The electrochemistry of these complexes was examined, and the semi-quinone complexes [Ru(CO)(2)L(2)(PHAQ-2H)](1+) and [Ru(CO)(dppe)(PBu(3))(PHAQ-2H)](1+) were generated by chemical oxidation. One example of an o-acylphenolate complex, HRu(CO)(PCy(3))(2)(PUR-H), is also reported. PMID- 11487344 TI - Novel silicate anion: Si(8)O(22)(12-). Hydrothermal synthesis and X-ray powder structure of three new niobium silicates. AB - A sodium niobium(V) tetrasilicate, Na(2)H(NbO)Si(4)O(11).1.25H(2)O (1), was synthesized hydrothermally at 190 degrees C from a mixture of silicic acid, NaOH, NbCl(5), and H(2)O(2) and added hydrochloric acid. The successive treatment of 1 with nitric acid yielded HNb(H(2)O)Si(4)O(11).H(2)O (2). Contact of 2 with cesium hydroxide solutions converted it to the partially exchanged Cs(+) phase Cs(0.66)H(0.33)Nb(H(2)O)Si(4)O(11) (3). The three structures were solved from X ray powder diffraction data. All compounds crystallize in the monoclinic space group P2(1)/m with Z = 2, containing the Si(8)O(22)(12-) anion. This new silicate anion type is related to the Si(4)O(11)(6-) unit, which is present in the amphibole series. The difference between both anion types lies in the chain periodicity: two for amphiboles or four for the new niobium silicates. These niobium silicates have framework structures enclosing tunnels in which the alkali cations reside. PMID- 11487345 TI - Synthesis and DNA binding of novel water-soluble cationic methylcobalt porphyrins. AB - Organocobalt derivatives of tetracationic water-soluble porphyrins are difficult to prepare via the typical reductive alkylation of the Co(II)(por) (porH(2) = porphyrin ligand). None have been reported. The problem may arise because the porphyrin core is made relatively electron poor by the positively charged peripheral groups. We have circumvented this problem by using the [Co(III)(NH(3))(5)CH(3)](2+) reagent, which inserts the Co(III)-CH(3) moiety directly into porH(2) in water under basic conditions. The method afforded two new [CH(3)Co(por)](4+) derivatives, [CH(3)CoTMpyP(4)](4+) and [CH(3)CoTMAP](4+), where [TMpyP(4)](4+) and [TMAP](4+) are the coordinated, NH-deprotonated forms of meso-tetrakis(N-methyl-4-pyridiniumyl)porphyrin and meso-tetrakis(N,N,N trimethylaniliniumyl)porphyrin, respectively. The binding of the two new [CH(3)Co(por)](4+) cations to DNA and to the synthetic DNA polymers [poly(dA dT)](2) and [poly(dG-dC)](2) was studied. Using published criteria by which changes in DNA viscosity and in the visible and CD spectra in the Soret region can be used to assess DNA binding, we conclude that both are outside binders. A large hypochromicity of the Soret bands of the [CH(3)Co(por)](4+) cations observed upon outside binding to DNA may indicate a high degree of self-stacking. The visible absorption and CD spectra of the [CH(3)Co(por)](4+) cations in the presence of 1:1 mixtures of [poly(dA-dT)](2) and [poly(dG-dC)](2) are nearly identical to those with [poly(dA-dT)](2) alone and are very different from those of [poly(dG-dC)](2) alone. Thus, both cations show a high preference for outside binding at AT-rich over GC-rich DNA sites. Upon binding of each of the [CH(3)Co(por)](4+) cations to all of the DNA polymers, the Soret bands exhibit blue shifts, whereas the Soret bands of the corresponding [(H(2)O)(2)Co(por)](5+) cations exhibit red shifts. The blue shifts strongly suggest that the [CH(3)Co(por)](4+) cations, particularly [CH(3)CoTMAP](4+), become five coordinate forms to some extent on DNA binding; this result is the first good evidence for the presence at equilibrium of five-coordinate CH(3)Co(III)(N(4)) forms in water. PMID- 11487346 TI - Synthesis and structural characterization of tin(II) and zinc(II) derivatives of cyclic alpha-hydroxyketones, including the structures of Sn(maltol)(2), Sn(tropolone)(2), Zn(tropolone)(2), and Zn(hinokitiol)(2). AB - Zinc(II) and tin(II) derivatives of maltol (Hmalt), ethylmaltol (HEtmalt), tropolone (Htrop), hinokitiol (Hhino), and kojic acid (Hkoj) have been prepared and characterized, and the crystal structures of M(trop)(2) (M = Zn, Sn), Zn(hino)(2).EtOH, and Sn(malt)(2) have been determined. The Zn(trop)(2) is a polymeric structure in which tropolone has both a bridging and chelating role; zinc(hino)(2) crystallizes as an ethanol adduct of which the structure is a dimeric fragment of the Zn(trop)(2) polymer and in which each metal is "capped" by a molecule of alcohol. The tin complexes are notably air-stable despite adopting monomeric pseudo-trigonal-bipyramidal structures (SnO(4)E; E is a stereochemically active lone electron pair) in which the ligands only chelate a single metal center. PMID- 11487347 TI - Influence of extrinsic factors on electron transfer in a mixed-valence Fe(2+)/Fe(3+) complex: experimental results and theoretical considerations. AB - The crystal structure of the mixed-valence complex (NEt(4))[Fe(2)(salmp)(2)].xMeCN(crystal) (x = 2,3) [1].xMeCN(crystal) was determined at temperatures between 153 and 293 K. The complex shows distinct Fe(2+) and Fe(3+) sites over this temperature interval. Variable temperature Mossbauer spectra confirm the valence-localized character of the complex. In contrast, spectroscopic investigation of powder samples generated from [1].xMeCN(crystal) indicate the presence of a valence-averaged component at temperatures above 150 K. To elucidate this apparent contradiction we have conducted a variable-temperature Mossbauer investigation of different forms of 1, including [1].xMeCN(crystal), [1].2DMF(crystal), [1].yMeCN(powder), and solution samples of 1 in acetonitrile. The low-temperature Mossbauer spectra of all forms are virtually identical and confirm the valence-localized nature of the S = (9)/(2) ground state. The high-temperature spectra reveal a subtle control of electron hopping by the environment of the complexes. Thus, [1].xMeCN(crystal) has valence-localized spectra at all explored temperatures, [1].2DMF(crystal) exhibits a complete collapse into a valence-averaged spectrum over a narrow temperature range, the powder exhibits partial valence averaging over a broad temperature interval, and the solution sample shows at 210 K the presence of a valence-averaged component in a minor proportion. The spectral transformations are characterized by a coexistence of valence-localized and valence-averaged spectral components. This phenomenon cannot be explained by intramolecular electron hopping between the valence-localized states Fe(A)(2+)Fe(B)(3+) and Fe(A)(3+)Fe(B)(2+) in a homogeneous ensemble of complexes, but requires relaxation processes involving at least three distinguishable states of the molecular anion. Hopping rates for [1].2DMF(crystal) and [1].yMeCN(powder) have been determined from spectral simulations, based on stochastic line shape theory. Analysis of the temperature dependences of the transfer rates reveals the existence of thermally activated processes between (quasi) degenerate excited states in both forms. The preexponential factors in the rate law for the hopping processes in the [1].yMeCN(powder) and [1].2DMF(crystal) differ dramatically and suggest an important influence of the asymmetry of the complex environment (crystal) on intramolecular electron hopping. The differences between the spectra for the crystalline sample [1].xMeCN and those for powders generated under vacuum from these crystals indicate that solvate depletion has a profound effect on the dynamic behavior. Finally, two interpretations for the three states involved in the relaxation processes in 1 are given and critically discussed (salmp = bis(salicyledeneamino)-2-methylphenolate(3-)). PMID- 11487348 TI - Synthesis and characterization of the first examples of perfluoroalkyl substituted trialkyloxonium salts, [(CH3)2OCF3]+[Sb2F11]- and [(CH3)2OCF(CF3)2]+[Sb2F11]-. AB - In the superacidic HF/SbF(5) system, methyl trifluoromethyl ether forms at -78 degrees C the new tertiary oxonium salt [(CH(3))(2)OCF(3)](+)[Sb(2)F(11)](-), which was characterized by Raman and multinuclear NMR spectroscopy and its crystal structure. The same oxonium salt was also obtained by methylation of CH(3)OCF(3) with CH(3)F and SbF(5) in HF solution at -30 to -10 degrees C. Replacement of one methyl group in the trimethyloxonium cation by the bulkier and more electronegative trifluoromethyl group increases the remaining O-CH(3) bond lengths by 0.037(1) A and the sum of the C-O-C bond angles by about 4.5 degrees. Methylation of CH(3)OCF(CF(3))(2) with CH(3)F in HF/SbF(5) solution at -30 degrees C produces [(CH(3))(2)OCF(CF(3))(2)](+)[Sb(2)F(11)](-). The observed structure and vibrational and NMR spectra were confirmed by theoretical studies at the B3LYP/6-311++G(2d,2p) and the MP2/6-311++G(2d,p) levels. PMID- 11487349 TI - Interaction of ferrocenoyl-dipeptides with 3-aminopyrazole derivatives: beta sheet models? A synthetic, spectroscopic, structural, and electrochemical study. AB - The use of 3-aminopyrazole derivatives as beta-sheet templates is investigated using a series of ferrocenoyl (Fc)-dipeptides (Fc-Gly(2)-OEt, Fc-Ala(2)-OBzl, Fc Leu-Phe-OMe, Fc-Val-Phe-OMe, Fc-Phe(2)-OMe, Fc-Leu(2)-OMe, Fc-Val(2)-OMe). The synthesis and full characterization are reported. The solid-state structures of Fc-Gly(2)-OMe and Fc-Leu-Phe-OMe show extensive hydrogen bonding of the podand peptide substituents, resulting in the formation of supramolecular Fc-dipeptide assemblies. For Fc-Gly(2)-OMe, this can be described as a parallel beta-sheet, whereas intermolecular interactions in Fc-Leu-Phe-OMe result in the formation of supramolecular helical structures. The saturation titrations of Fc-dipeptides with 3-amino-5-methylpyrazole (3-AMP) and 3-trifluoroacetylamido-5-methylpyrazole (3-TFAc-AMP) show a 1:1 interaction of the Fc-peptide with the aminopyrazole derivatives. IR measurements in solution confirm binding to the top face of the Fc-dipeptide and the involvement of the Fc-C=O and the ester C=O groups in establishing H-bonding interactions with the 3-TFAc-AMP. However, binding constants in chloroform are low and range from 8 to 27 M(-1), which correspond to binding energies of 5-7 kJ mol(-1). In higher polarity solvents, such as acetonitrile or acetone, the binding constants are below 5 M(-1), emphasizing the limited utility of 3-AMP derivatives as beta-sheet templates. Electrochemical measurements confirm the weak interactions between the various Fc-dipeptides and 3-TFAc-AMP. Typical shifts in the redox potential of the Fc moiety are in the range 0-20 mV. Attempts to modify 3-AMP at the 3-position by carbodiimide coupling with amino acid derivatives and, thus, enhance the binding to the Fc peptides resulted in 2-amino acid substituted 3-AMP derivatives. Substitution at the 2-position blocks the binding site, and no interactions with Fc-dipeptides are observed. PMID- 11487350 TI - Synthesis and coordination properties of new bis(phosphinomethyl)pyridine N,P,P' trioxides. AB - In a search for more hydrocarbon solvent soluble derivatives of the parent ligand, 2,6-[Ph(2)P(O)CH(2)](2)C(5)H(3)NO (1a), a series of new ligands, 2,6 [R(2)P(O)CH(2)](2)C(5)H(3)NO [R = Bz (1b); Tol (1c); Et (1d); Pr (1e); Bu (1f); Pn (1g); Hx (1h); Hp (1i); and Oct (1j)] and 2,6-[RR'P(O)CH(2)](2)C(5)H(3)NO [R = Ph, R' = Bz (2a); R = Ph, R' = Me (2b); R = Ph, R' = Hx (2c); R = Ph, R' = Oct (2d)], have been prepared by either Arbusov or Grignard substitutions on 2,6 bis(chloromethyl)pyridine followed by N-oxidation. The new ligands have been characterized by spectroscopic methods, and their coordination chemistry with selected lanthanide ions has been surveyed. Several 1:1 and 2:1 ligand/metal complexes have been isolated, and single-crystal X-ray diffraction analyses for Nd(2a)(NO(3))(3), Er(2a)(NO(3))(3), Yb(1d)(NO(3))(3), and [Nd(1c)(2)](NO(3))(3) are described. The new structural data are discussed in relation to the structures of complexes formed by 1a. PMID- 11487351 TI - Complexation studies of iodides of trivalent uranium and lanthanides (Ce and Nd) with 2,2'-bipyridine in anhydrous pyridine solutions. AB - In anhydrous pyridine solution at 294 K, U(III) and Ce(III) triiodides were found to form both 1:1 (ML) and 1:2 (ML(2)) complexes with bipyridine (bipy = L) while Nd(III) triodide formed only a 1:2 complex. The 1:3 (ML(3)) complexes were identified at low temperature with a large excess of L. Conductometry measurements showed for U(III) a large increase in the conductivity when increasing the molar ratio L:U. The complex UL(2) was found to be a 1:1 electrolyte and the species UI(2)(+) was more reactive toward L in comparison with UI(3). For Ce(III) and Nd(III), MI(2)(+) and MI(3) present about the same affinity for L. The stability of the complexes is limited, and U(III) possesses a slightly higher affinity for bipy than the trivalent lanthanides. Interestingly, a preference for the formation of ML(2) complex was shown for all the studied M(III) ions. The driving force for complex formation was always the enthalpy, and, surprisingly for a bidendate ligand (bipy), no favorable entropy contribution to complex formation was observed. The X-ray crystal structures of [CeI(3)(bipy)(2)(py)](4).5py.bipy and UI(3)(bipy)(2)(py).2py were determined. The structures of the molecules MI(3)(bipy)(2)(py) are almost identical for U and Ce. The mean M(III)-N(bipy) bond distances are equal to 2.67(3) A for Ce(III) and 2.65(4) A for U(III). The slightly smaller M(III)-N(bipy) distances observed for U(III) would reflect a slightly more important covalent character of the U(III) N(bipy) bonds, in agreement with the slightly better affinity of U(III) than Ce(III) or Nd(III) toward bipy observed in solution and with the fact that the enthalpy is the driving force for complex formation. PMID- 11487352 TI - Kinetics and mechanisms of aqueous ozone reactions with bromide, sulfite, hydrogen sulfite, iodide, and nitrite ions. AB - Reactions of ozone with Br(-), SO(3)(2-), HSO(3)(-), I(-), and NO(2)(-), studied by stopped-flow and pulsed-accelerated-flow techniques, are first order in the concentration of O(3)(aq) and first order in the concentration of each anion. The rate constants increase by a factor of 5 x 10(6) as the nucleophilicities of the anions increase from Br(-) to SO(3)(2-). Ozone adducts with the nucleophiles are proposed as steady-state intermediates prior to oxygen atom transfer with release of O(2). Ab initio calculations show possible structures for the intermediates. The reaction between Br(-) and O(3) is accelerated by H(+) but exhibits a kinetic saturation effect as the acidity increases. The kinetics indicate formation of BrOOO(-) as a steady-state intermediate with an acid-assisted step to give BrOH and O(2). Temperature dependencies of the reactions of Br(-) and HSO(3)(-) with O(3) in acidic solutions are determined from 1 to 25 degrees C. These kinetics are important in studies of annual ozone depletion in the Arctic troposphere at polar sunrise. PMID- 11487353 TI - Chemistry of 2,4,6-trimercapto-1,3,5-triazine (TMT): acid dissociation constants and group 2 complexes. AB - The acid dissociation constants of 2,4,6-trimercaptotriazine (H(3)TMT, 1) were determined and now can be employed in the preparation of complexes having specific M-TMT (M = divalent metal; TMT = 2,4,6-trimercapto-1,3,5-triazinide, C(3)N(3)S(3)(3-)) ratios. For example, the combination of H(3)TMT (1) with Mg(OH)(2) at pH 7.1 leads to the crystallization of Mg(H(2)TMT)(2).6H(2)O (4). With the appropriate pH adjustment, the contiguous series of compounds Ba(3)(TMT)(2).8H(2)O (3), Ba(H(2)TMT)(2).7H(2)O (5), and BaHTMT.3H(2)O (6) can be isolated. The compounds were characterized by mp, IR, TGA, elemental analysis, and, in the cases of 4, 5, and 6, crystallography. The comparison of 4 with 5 and 6 offers an interesting view of the difference in hard and soft bonding with TMT. In the saltlike Mg structure of 4, there is extensive hydrogen bonding, but in the Ba structures, 5 and 6, covalent Ba-S bonding dominates. PMID- 11487354 TI - Pentanuclear homoleptic M(5)L(6) (M = Mn(II), Co(II), Zn(II)) complexes formed by strict self-assembly. AB - A series of trigonal bipyramidal pentanuclear complexes involving the alkoxo diazine ligands poap and p3oap, containing the M(5)[mu-O](6) core is described, which form by a strict self-assembly process. [Co(5)(poap H)(6)](ClO(4))(4).3H(2)O (1), [Mn(5)(poap-H)(6)](ClO(4))(4).3.5CH(3)OH.H(2)O (2), [Mn(5)(p3oap-H)(6)](ClO(4))(4).CH(3)CH(2)OH.3H(2)O (3), and [Zn(5)(poap H)(6)](ClO(4))(4).2.5H(2)O (4) are homoleptic pentanuclear complexes, where there is an exact match between the coordination requirements of the five metal ions in the cluster, and the available coordination pockets in the polytopic ligand. [Zn(4)(poap)(poap-H)(3)(H(2)O)(4)] (NO(3))(5).1.5H(2)O (5) is a square [2 x 2] grid with a Zn(4)[mu-O](4) core, and appears to result from the presence of NO(3), which is thought to be a competing ligand in the self-assembly. X-ray structures are reported for 1, 4, and 5. 1 crystallized in the monoclinic system, space group P2(1)/n with a = 13.385(1) A, b = 25.797(2) A, c = 28.513(3) A, beta = 98.704(2) degrees, and Z = 4. 4 crystallized in the triclinic system, space group P1 with a = 13.0897(9) A, b = 18.889(1) A, c = 20.506(2) A, alpha = 87.116(1) degrees, beta = 74.280(2) degrees, gamma = 75.809(2) degrees, and Z = 2. 5 crystallized in the monoclinic system, space group P2(1)/n with a = 14.8222(7) A, b = 21.408(1) A, c = 21.6197(9) A, beta = 90.698(1) degrees, and Z = 4. Compounds 1-3 exhibit intramolecular antiferromagnetic coupling. PMID- 11487355 TI - Saccharinate as a versatile polyfunctional ligand. Four distinct coordination modes, misdirected valence, and a dominant aggregate structure from a single reaction system. AB - The reaction system consisting of copper, saccharinate, and the auxiliary ligands H(2)O, PPh(3), and NH(3) produces a sequence of compounds in which saccharinate is coordinated to copper in four distinct manners. The complex trans [Cu(sacch)(2)(H(2)O)(4)] (2) (produced by thermal dehydration of trans [Cu(sacch)(2)(H(2)O)(4)].2H(2)O (1)) reacts with triphenylphosphine in CH(2)Cl(2) to produce any or all of three Cu(I) complexes, depending upon conditions. The three Cu(I) compounds are Cu(sacch)(PPh(3))(3) (3), in which saccharinate binds to copper through the carbonyl group of the ligand, Cu(sacch)(PPh(3))(2) (4), in which sacch binds to Cu through its charge-bearing nitrogen atom; and [Cu(sacch)(PPh(3))](2) (5), a dinuclear complex in which saccharinate bridges two Cu centers through its imidate nitrogen and carbonyl oxygen atoms. Complexes 3-5 can be isolated individually, although in solution they exist in a complex equilibrium which has been examined by NMR spectroscopy. Each of the three Cu(I) products reacts with NH(3) in CH(2)Cl(2) solution to produce trans [Cu(sacch)(2)(NH(3))(4)] (6), an unstable Cu(II) complex that exhibits misdirected valence at the Cu-N(sacch) bond. Complex 6 evolves spontaneously to [Cu(sacch)(NH(3))(4)](sacch).H(2)O (7), which in the solid state is dominated by a supramolecular aggregate of two formula units, linked by hydrogen bonding in which the water molecule plays a central role. Alternative pathways exist to several of the products. The X-ray crystal structure analyses of 3-7 are reported and establish the coordination modes of saccharinate, the misdirected valence in 6, and the supramolecular aggregation in 7. The structure analysis of 7 by single crystal neutron diffraction is reported and together with the previously reported neutron structure analysis of 1 establishes the substitution of the auxiliary ligand H(2)O by NH(3) in the Cu(II) products. PMID- 11487356 TI - Zeolite-based organized molecular assemblies. photophysical characterization and documentation of donor oxidation upon photosensitized charge separation. AB - An organized molecular assembly composed of two ruthenium polypyridine complexes, Ru(bpy)(2)(bpz)(2+) and Ru(bpy)(2)(H(2)O)(2)(2+) (where bpy = 2, 2'-bipyridine and bpz = 2, 2'-bipyrazine), has been prepared in adjacent supercages of Y zeolite. This material has been characterized by diffuse reflectance, electronic absorption, electronic emission, and resonance Raman (RR) spectroscopy, as well as lifetime measurements. The spectral results confirm the identity of the entrapped complexes and resonance Raman measurements show that the relative concentrations of the two complexes within the zeolite particles are identical. A dramatic decrease in emission intensity observed for the adjacent cage assembly, relative to that observed for an appropriate reference material composed of a mixture of zeolite particles containing the separated complexes, indicates strong interaction between the adjacent complexes which provides an additional nonradiative decay pathway. The excited state lifetime measurements implicate a very short-lived component, dominating the decay curve at early times, which is most reasonably attributed to excited-state electron-transfer quenching of the adjacent cage pair. More importantly, analysis of diffuse reflectance spectra acquired during selective (sensitizer) irradiation of a sample of this material, wherein the remaining cages are filled with a suitable acceptor (MV(2+)), provides direct evidence for oxidation of the Ru(bpy)(2)(H(2)O)(2)(2+) donor complex, confirming the targeted synergy of the adjacent cage assembly. PMID- 11487357 TI - Synthesis and crystal structure of Ae(2)LiInGe(2) (Ae = Ca, Sr): new Zintl phases with a layered silicate-like network. AB - Two new Zintl phases Ae(2)LiInGe(2) (Ae = Ca 1; Sr 2) were obtained from stoichiometric reactions of the pure elements in sealed Nb tubing at 1000-1050 degrees C. The isomorphous polar intermetallic phases crystallize in the orthorhombic space group Pnma, with cell constants of a = 7.2512(7), b = 4.4380(5), and c = 16.902(1) A for compound 1, and a = 7.5033(8), b = 4.6194(5), and c = 17.473(2) A for compound 2. The crystal structure can be derived from the vertex-sharing of InGe(4/2) tetrahedral units that form "corrugated" sheets normal to the crystallographic c-axis. Calcium and lithium atoms act as "spacers" that effectively separate the anionic [InGe(2)](5-) layers. The layered anionic substructure is similar to those exhibited by layered metal oxides, sulfides, and silicates. The connectivity of the tetrahedral building unit, [InGe(4/2)](5-), is analogous and isoelectronic to the silicate [SiO(4/2)] unit. PMID- 11487358 TI - Clarification of the crystal structure of [(HC(Ph2PO)3)2Cu](ClO4)2.2H2O. PMID- 11487359 TI - A mimic molecule of blue copper protein active site [(-)-sparteine N,N'](maleonitriledithiolato-S,S')copper(II). PMID- 11487360 TI - Synthesis and characterization of the GeO(4)Al(12)(OH)(24)(OH2)(12)(8+) polyoxocation. PMID- 11487361 TI - The first simple, donor-free salt of the Sb(OTeF(5))(6)(-) anion: synthesis, structure, characterization, and thermochemistry of Cs[Sb(OTeF5)6]. PMID- 11487362 TI - Syntheses and structures of bis(azido)- and bis(tert-butoxy)cyclodistibazanes. PMID- 11487363 TI - Electronic structure of low-spin ferric chlorins: characterization of bis(dimethylphenylphosphine)(meso-tetraphenylchlorinato)iron(III) triflate. PMID- 11487364 TI - Modeling the anomalous coordination chemistry of Cr(III) biscyclam. PMID- 11487365 TI - A silver-linked supramolecular cluster encapsulating a cesium cation. PMID- 11487366 TI - Malaria diagnosis and treatment under the strategy of the integrated management of childhood illness (IMCI): relevance of laboratory support from the rapid immunochromatographic tests of ICT Malaria P.f/P.v and OptiMal. AB - The algorithm developed for the integrated management of childhood illness (IMCI) provides guidelines for the treatment of paediatric malaria. In areas where malaria is endemic, for example, the IMCI strategy may indicate that children who present with fever, a recent history of fever and/or pallor should receive antimalarial chemotherapy. In many holo-endemic areas, it is unclear whether laboratory tests to confirm that such signs are the result of malaria would be very relevant or useful. Children from a holo-endemic region of Tanzania were therefore checked for malarial parasites by microscopy and by using two rapid immunochromatographic tests (RIT) for the diagnosis of malaria (ICT Malaria P.f/P.v and OptiMal. At the time they were tested, each of these children had been targeted for antimalarial treatment (following the IMCI strategy) because of fever and/or pallor. Only 70% of the 395 children classified to receive antimalarial drugs by the IMCI algorithm had malarial parasitaemias (68.4% had Plasmodium falciparum trophozoites, 1.3% only P. falciparum gametocytes, 0.3% P. ovale and 0.3% P. malariae). As indicators of P. falciparum trophozoites in the peripheral blood, fever had a sensitivity of 93.0% and a specificity of 15.5% whereas pallor had a sensitivity of 72.2% and a specificity of 50.8%. The RIT both had very high corresponding sensitivities (of 100.0% for the ICT and 94.0% for OptiMal) but the specificity of the ICT (74.0%) was significantly lower than that for OptiMal (100.0%). Fever and pallor were significantly associated with the P. falciparum asexual parasitaemias that equalled or exceeded the threshold intensity (2000/microl) that has the optimum sensitivity and specificity for the definition of a malarial episode. Diagnostic likelihood ratios (DLR) showed that a positive result in the OptiMal test (DLR = infinity) was a better indication of malaria than a positive result in the ICT (DLR = 3.85). In fact, OptiMal had diagnostic reliability (0.93) which approached that of an ideal test and, since it only detects live parasites, OptiMal is superior to the ICT in monitoring therapeutic responses. Although the RIT may seem attractive for use in primary health facilities because relatively inexperienced staff can perform them, the high cost of these tests is prohibitive. In holo-endemic areas, use of RIT or microscopical examination of bloodsmears may only be relevant when malaria needs to be excluded as a cause of illness (e.g. prior to treatment with toxic or expensive drugs, or during malaria epidemics). Wherever the effective drugs for the first-line treatment of malaria are cheap (e.g. chloroquine and Fansidar), treatment based on clinical diagnosis alone should prove cost-saving in health facilities without microscopy. PMID- 11487367 TI - Apparent drug failure following artesunate treatment of Plasmodium falciparum malaria in Freetown, Sierra Leone: four case reports. AB - Four cases of Plasmodium falciparum malaria who presented in Sierra Leone in November-December 2000 apparently failed to respond to treatment with artesunate. Three (75%) of the cases fulfilled the World Health Organization's criteria for late treatment failure. Although artesunate ranks only sixth as the first-line drug used by clinicians for the treatment of uncomplicated malaria in Sierra Leone, it is widely sold over the counter in pharmacies in the country. The indiscriminate and injudicious use of artesunate among the Sierra Leonean population is likely to increase the level and frequency of resistance among the local strains of P. falciparum. It is recommended that artesunate be reserved for patients who fail to respond to treatment with another of the antimalarial drugs available. Even then, the artesunate should preferably be used in combination with other, longer-acting antimalarial drugs, to slow the development of further resistance. PMID- 11487368 TI - Modulation of the humoral response to repeat and non-repeat sequences of the circumsporozoite protein of Plasmodium vivax using novel adjuvant and delivery systems. AB - In the use of sub-unit vaccines, it is important to identify the protective epitopes and to generate the optimal immune response by using appropriate immuno modulatory adjuvants and/or delivery systems. The main aim of the present study was to generate an MHC-non-restricted immune response against one promising vaccine candidate, the circumsporozoite protein (CSP) of Plasmodium vivax. Four synthetic peptides were chosen: three repeat-region sequences (AA, DA and ANG) and a putative T-cell epitope extended from a conserved region (region II) containing a hepatocyte-binding region (HBP). The humoral response against each peptide was studied in outbred mice and three strains of inbred mice (with different genetic backgrounds). Delivery of each peptide in microspheres or inclusion of a bio-active casein-fragment analogue as adjuvant with alum/liposome delivery considerably enhanced the humoral response against the peptide (when compared with the response to the peptide delivered in alum alone). The maximal immune response was observed when the peptide was delivered in microspheres, with no booster doses required; the antibodies raised against peptide delivered with adjuvant or in modulatory delivery vehicles had two-to five-fold lower binding affinities. The predominant IgG isotypes elicited using microspheres or adjuvant with alum/liposome delivery were IgG(2a)/IgG(2b) and/or IgG(1). Importantly, conjugation of HBP to the B-cell repeat peptides increased the titres of peptide specific antibodies, especially of antibodies against the supposedly cryptic HBP. Delivery of a mix of all four peptides in microspheres elicited an intense immune response in outbred mice, indicating that such a delivery system efficiently presents the peptides to the immune effector cells. That antibodies in the anti peptide sera bound strongly to air-dried sporozoites of P. vivax was confirmed by immunofluorescence. The present results, based on the use of individual peptides or a conjugate or cocktail of the peptides, highlight the utility of the casein fragment analogue as an adjuvant, when used with alum/liposome delivery, and also demonstrate the potential of microspheres as a single-shot delivery system for sub-unit peptides. PMID- 11487369 TI - Immunological selection for Leishmania (Viannia) braziliensis antigens. AB - Comparative ELISA and selective immunoblotting procedures were used in attempts to identify differential serological indicators of infection with the Leishmania (Viannia) braziliensis complex, infection with the L. braziliensis species, and therapeutic cure of localized or mucocutaneous leishmaniasis (LCL or MCL). Although mean ELISA absorbance values were significantly higher for MCL sera than for LCL sera, absorbance could not be used as a reliable indicator of the clinical form of disease. Immunoblotting profiles were similar with sera from MCL and LCL. Pre-adsorption with heterologous trypanosomatid antigens indicated that recognition of antigens of about 56, 60, 66, 72, 88 and 110 kDa might be specific to the subgenus Viannia. In two-colour, sequential, dual ELISA-based immunoblotting, no antigens recognized only by sera from MCL patients were detected. After glucantime therapy, immunoblotting profiles with LCL sera were reduced both in intensity and in the range of antigens detected; a 104-kDa antigen was newly detected with post-treatment LCL sera. Overall, the results show the value of differential immunological detection strategies and support the close relationship between species of the subgenus Viannia but fail to indicate a prognostic antigen for MCL. PMID- 11487370 TI - Involvement of women in community-directed treatment with ivermectin for the control of onchocerciasis in Rukungiri district, Uganda: a knowledge, attitude and practice study. AB - A study of knowledge, attitudes and practice was carried out in the Rukungiri district of Uganda, in order to investigate the involvement of women in community directed treatment with ivermectin (CDTI), for the control of onchocerciasis. The data analysed came from interviews with 260 adult women (one from each of 260 randomly-selected households in 20 onchocerciasis-endemic communities), community informants, and participatory evaluation meetings (PEM) in eight communities. The women who had been treated with ivermectin in 1999 generally had more knowledge of the benefits of taking ivermectin, were more likely to have attended the relevant health-education sessions and were more involved in community decisions on the method of ivermectin distribution than the women who had not received ivermectin in that year. There were fewer female community-directed health workers (CDHW) than male CDHW in the communities investigated. The reasons for not attending health-education sessions, not participating in community meetings concerning the CDTI, and the reluctance of some women to serve as CDHW were investigated. The most common reasons given were domestic chores, a reluctance to express their views in meetings outside their own kinship group, suspicions that other women might take advantage of them, and a lack of interest. Most of the women interviewed (as well as other community members) felt that there were relatively few women CDHW. The women attributed this to a lack of interaction and trust amongst themselves, which resulted in more men than women being selected as CDHW. The rest of the community members were not against women working as CDHW. It is recommended that communities be encouraged to select women to serve as CDHW in the CDTI, and that the performances of male and female CDHW be compared. PMID- 11487371 TI - Relationships between the prevalence and intensity of Loa loa infection in the Central province of Cameroon. AB - Ivermectin treatment may induce marked adverse effects in those harbouring > 8000 Loa microfilariae (mff)/ml of blood, individuals with > 30 000 Loa mff/ml being at risk of developing serious neurological reactions. It is thus necessary to delineate the geographical areas where such responses may occur. To determine if the simple measure of prevalence of Loa microfilaraemia would be appropriate to identify the communities at risk, the relationships between prevalence and intensity of Loa microfilaraemia were investigated in 67 villages in Cameroon. The prevalence recorded in the adult population was found to be closely related to each of the indicators of infection intensity investigated. For example, when the prevalences of Loa microfilaraemia in adults were 20%, 30% and 40%, approximately 5%, 9% and 16% of the adults had microfilarial loads exceeding 8000 mff/ml, respectively; the corresponding percentages of adults with > 30 000 mff/ml were about 1%, 3% and 5%-6%. Thus it seems that, in areas where loiasis is co-endemic, the monitoring procedure during large-scale ivermectin treatments for the control of onchocerciasis only needs to be strengthened in those communities where the prevalence of Loa microfilaraemia in adults exceeds 20%. PMID- 11487372 TI - Dysentery in patients infected with human immunodeficiency virus in Zimbabwe: an emerging role for Schistosoma mansoni and Escherichia coli O157? AB - Information on bloody diarrhoea in HIV-positives is scarce. A prospective study was therefore performed, in Zimbabwe, to determine and compare the pathogens associated with bloody diarrhoea in 25 antiretroviral-naive HIV-infected patients and 15 non-HIV-infected patients. Stool cultures and colonic biopsies were performed. Shigella was isolated from 18 (45%) of the subjects, Schistosoma mansoni from eight (16%), Escherichia coli H7:O157 from three (8%) and Campylobacter jejunii from two (5%). There was no evidence of Salmonella, Entamoeba histolytica or cytomegalovirus infection. Shigella dysenteriae type-1 occurred more often in the HIV-negatives than the HIV-positives (P = 0.02). Although HIV-associated bloody diarrhoea in Zimbabwe appears to be most frequently caused by Shigella, it may also be commonly the result of infection with Sc. mansoni or shiga-toxin-producing E. coli. A larger study specifically to examine the role of Sc. mansoni and E. coli O157 is warranted. PMID- 11487373 TI - Spatio-temporal genetic variability in the schistosome intermediate host Biomphalaria pfeifferi. AB - Genetic variation among temporally and spatially defined populations of the schistosome intermediate host, Biomphalaria pfeifferi, was analysed using RAPD markers. Snails were collected from each of eight sites that were 1-20 km apart, on four occasions, over a 2-year period. High levels of genetic variability were found within populations, between populations and between collections. Spatial differentiation and intrapopulation similarity were highest during periods of drought and were lowest following heavy rainfall. Temporal variability, consistent with founder effects, genetic drift and/or differential parasite pressure, was of similar or greater magnitude to that of the spatial variability. The results not only demonstrate that spatio-temporal sampling is necessary to understand the population genetics of short-lived species in variable habitats, but also provide empirical data to support current theoretical models of host parasite co-evolution. PMID- 11487374 TI - The antileishmanial properties of surface-modified, primaquine-loaded nanocapsules tested against intramacrophagic Leishmania donovani amastigotes in vitro. PMID- 11487375 TI - High prevalence of Wuchereria bancrofti infection among Myanmar migrants in Thailand. PMID- 11487376 TI - The acarine fauna in dust samples from domestic pantries in southern Brazil. PMID- 11487378 TI - The platelet release reaction: granules' constituents, secretion and functions. AB - Although anucleated, blood platelets are highly organized cells rich in different types of organelles. Three specific granule populations store different types of constituents, some of which are at high concentrations. Platelets thus transport some specific compounds through the whole body. During circulation, platelets are reactive to various stimuli and release the materials stored in the specific granules. This 'release reaction' is an important step of primary haemostasis. Energy and messengers required for platelet reactivity are provided by mitochondria and the dense tubular system. Each granule population has specific properties concerning both the structure and the role played by the released constituents. Dense granules contain small non-protein molecules that are secreted to recruit other platelets. alpha-Granules contain large adhesive and healing proteins. Lysosomes contain hydrolases able to eliminate the circulating platelet aggregate. The extrusion of storage granules' content to the platelet's environment occurs according to regulated secretion events: movements of granules, apposition and fusion of granule and plasma membranes. Typical platelet disorders resulting from a storage granule abnormality are referred to as a storage pool defect and are characterized by a prolonged bleeding time. PMID- 11487379 TI - A single (investigator)-blind randomised control trial comparing the effects of quinapril and nifedipine on platelet function in patients with mild to moderate hypertension. AB - The effect of quinapril and nifedipine on platelet aggregation, vascular endothelial function and coagulation system activity, was compared in a parallel group, investigator-blind study carried out on patients with mild to moderate hypertension but no other diseases or receiving medication which might affect platelet function, vascular endothelium or coagulation. Forty patients (two groups of 20 patients each) and 20 control subjects were recruited. Patients were randomised to receive either quinapril or nifedipine retard and the dose escalated to control hypertension. Platelet aggregation studies were assessed serially and beta-thromboglobulin, angiotensin-converting enzyme (ACE), von Willebrand factor (vWF) coagulation factors VIIIc, XII and fibrinogen were measured at the beginning and end of the 12-week period. Blood pressure was adequately controlled in all patients in both groups. Platelet function was impaired in certain parameters (slope of the reaction with ADP and collagen and maximum aggregation with collagen) in the patient group compared to controls before treatment and this improved in patients on quinapril but not on nifedipine; likewise beta-thromboglobulin was higher in the patient group and fell significantly in the quinapril group but not those on nifedipine. Measurements of endothelial function and coagulation were normal before treatment and showed no alteration during the study, except in the expected fall in plasma ACE in the quinapril group. The results indicate that the ACE inhibitor, quinapril, has a beneficial effect on platelet function unlike the calcium channel blocker, nifedipine. PMID- 11487380 TI - Deaggregation is an integral component of the response of platelets to ADP in vitro: kinetic studies of literature and original data. AB - Adenosine diphosphate (ADP) is recognized as an important mediator of platelet aggregation. Transient aggregation at low (< or =1 microM), and sustained aggregation at higher ADP concentrations are consistently observed. Dissociation of platelet aggregates has been described and may explain the reversible component of the aggregation response. We hypothesized that the net aggregation response to ADP in vitro results from the concurrent activation of two opposing processes, aggregation and deaggregation. Different purinergic receptor subtypes may mediate these effects. To test this hypothesis and its generalizability, we performed a kinetic analysis of representative published ADP-induced aggregation responses supplemented with original data from our laboratory. A four-compartment kinetic model was used to estimate k(3), a rate constant of deaggregation. Two model-independent parameters, the magnitude of the aggregation response (DeltaOD) and the time to reach maximal aggregation (t(peak)) were also assessed. Greater sustained aggregation at higher ADP concentrations was consistently associated with increased DeltaOD and t(peak) but decreased k(3) values. These relationships were independent of type of platelet preparation or experimental conditions and not due to ADP receptor desensitization. Conversely, blockade of the P2Y(12) receptor subtype (ticlopidine, clopidogrel or 2-MeS-AMP) decreased DeltaOD and t(peak) but increased k(3) values. This supports the presence of active deaggregation which is decelerated by activation of the P2Y(12) receptor subtype. PMID- 11487381 TI - Altered megakaryocyte-platelet haemostatic axis in hypercholesterolaemia. AB - Hypercholesterolaemia is associated with accentuated platelet function. We assessed in a pilot study whether megakaryocytes (MK), the platelet precursor cell, were altered in subjects with primary hypercholesterolaemia and whether these changes were linked with platelet function. MK and platelet function were assessed in eight untreated patients with primary hypercholesterolaemia (total cholesterol, TC > 7 mmol/l), and 14 control subjects (TC < or = 7 mmol/l): MK ploidy (DNA content), size, granularity, and expression of the adhesion molecule GP IIIa, and platelet expression of GP IIIa, P selectin and CD 63, and RNA content, were each measured using flow cytometry; mean platelet volume, platelet count, plasma thrombopoietin, and cutaneous bleeding time were also assessed. Hypercholesterolaemic patients had increased MK ploidy, mean (SD) 22.9 N (2.3) vs. 20.8 N (1.6) (2P=0.021); platelet size, 10.6 fl (1.2) vs. 9.3 fl (0.7) (2P=0.006); and platelet expression of GP IIIa, 111.3 (18.9) vs. 92.0 (12.3) (2P=0.010), as compared with matched control subjects. Cutaneous bleeding time tended to be reduced in the hypercholesterolaemic patients, 364 s (136) vs. 483 s (165) (2P=0.11). No differences in MK size, granularity or GP IIIa expression, or platelet count, mass, RNA content, P selectin or CD 63 expression, or plasma thrombopoietin were seen. The data suggest that aspects of megakaryocytes and platelets are altered in hypercholesterolaemia, as has also been seen in other vascular risk factors states, including hypertension and diabetes mellitus. Furthermore, changes in megakaryocytes may have contributed to the altered platelet function seen here. Further study is now required to assess whether lipid lowering therapy "normalises" these changes in the megakaryocyte-platelet haemostatic axis. PMID- 11487382 TI - Studies on the improvement of leucodepletion performance of the Haemonetics MCS+ for production of leucodepleted platelet concentrate. AB - With the implementation of universal leucodepletion, an in-line, negatively charged LRF6H leucodepleting filter became an essential part of the Haemonetics MCS+ plateletpheresis system. A larger-scale (968) study using the standard protocol revealed a 2.79% leucodepletion failure rate (standard < 5 x 10(6) leucocytes per adult therapeutic dose). Factors influencing the efficacy of the filter were investigated. The pH of the filtrate was 7.0, the temperature 28 degrees C and filtration rate 80 ml/min. Reduction of the filtration rate to 30 ml/min (784 doses) reduced leucodepletion failure to 0.38%. Measurement of the leucocyte count, pre- and post-filtration of the platelet products, revealed that donations from 1% of donors contained substantially larger numbers of leucocytes in pre-filter samples (300-1500/microl) than in control samples (35-70/microl). This number tends to increase progressively with subsequent donations in these individuals, leading to leucodepletion failure, whilst peripheral leucocyte counts remain normal. The new continuous filtration protocol (version C) using a less impact filter LRF-XL and a lower (7 ml/min) head pressure was also effective but failure still occurred twice on one of the donors who persistently showed high pre-filter count. We conclude that leucodepletion failures in the Haemonetics system are related to both donor leucocyte (i.e., being light and non adherent) and operational/filter performance. PMID- 11487383 TI - A case of leucodepletion failure in Cobe spectra plateletpheresis: further studies on the identification of the potential cause. AB - A 54-year-old double dose donor failed leucodepletion by Cobe LRS for five sequential donations, without triggering the "machine failure alarm". All collections were uneventful but final products appeared lipaemic. The last two donations were studied: (i) by collecting the first half of each donation into one bag and the other half into a second bag. In both cases, the product in bag one was leucodepleted and that in bag two "failed" leucodepletion. (ii) By examining the filterability of the final products using an LRF6 filter. Leucodepletion was satisfactory but showed a slightly higher level of leucocytes as compared to control or lipaemic donations, possibly due to the presence of an interfering substance in the platelet product. (iii) By examining the cellular content of the LRS cone and comparing it with those obtained from lipaemic donations from other donors. Only the test cone contained a large mass of waxy white material. It is postulated that this material was gradually filling the cone and increasingly interfering with its leucodepleting function as the donation progressed. (iv) To monitor pre/post whole blood as well as the products for the presence of abnormal leucocyte subsets. The only difference was the presence of some extra abnormal dot plot. The intensity of these extra events which was reduced both upon donation and filtration. We conclude that, in this case, failure of leucodepletion was clearly donor rather than machine or process related. The implementation of this investigative study may help in the characterisation of the potential cause of leucodepletion failures. PMID- 11487384 TI - Whole blood microvolume laser scanning cytometry for monitoring resting and activated platelets. AB - Enumerating and phenotyping of platelets, resting and activated, from whole blood is important for both the identification and verification of many disease states. Microvolume laser scanning cytometry (MLSC) has been shown to be a simple method for enumerating and phenotyping peripheral blood cells. Here, the utility of MLSC, in conjunction with an anticoagulant containing platelet activation inhibitors, for simultaneously measuring platelet count, phenotype and responsiveness directly from non-fixed whole blood was examined. CTAD or EDTA anticoagulated blood was collected from five to 20 healthy volunteers, stained with fluorescence-labeled antibodies specific for platelet antigens, and run on an in-house modified MSLC device. MLSC was able to measure antigens CD9, CD29, CD36, CD41, CD42a, CD42b, and CD61 on platelets and determine an average of 2.3 x 10(5) +/- 7 x 10(4) platelets per microliter. Counts correlated well with those obtained from the Cell-Dyn 3500 (r(2)=0.84). Agreeing with previous data, less than 2% of platelets from peripheral blood of normal individuals expressed the activation markers CD62P or CD63. After in vitro thrombin activation, >93% of the platelets expressed activation markers. Data presented here shows the benefits of using MLSC in combination with platelet inhibitors to quantitate and phenotype platelets while maintaining a viable responsive state. PMID- 11487386 TI - 'Say no to sex; great idea but...'. PMID- 11487387 TI - Screening for sexually transmitted infections in children and adolescents in the United Kingdom: British Co-operative Clinical Group. AB - Our objectives were (1) to assess the number of young people aged under 16 years attending genitourinary medicine (GUM) departments in the UK in 1998; (2) to identify clinical activity and policy; (3) to determine the knowledge and training needs of healthcare professionals within GUM providing care for this client group. In July 1999 a questionnaire was circulated via the 18 regional British Co-operative Clinical Group (BCCG) representatives to the consultants in charge of all 197 main GUM departments in the UK. One hundred and sixty out of 197 (81%) completed questionnaires were returned and analysed. The reported number of under-16-year-olds attending in 1998 varied considerably between clinics; for females ranging from 0 to 256 and for males between 0 and 50, with a male to female ratio of 1:4.4. The majority of responding clinics, 139/160 (87%) had been involved in the screening of abused children/adolescents for sexually transmitted infections (STIs). Most clinics were prepared to screen for STI (86%), HIV test (79%) and assess contraceptive needs (50%) in this age group. Staff involved in care included health advisers (74), nurses (59), and doctors (138) in the responding clinics. Only 31/160 clinics (19%) had a written policy for the management of children/adolescents attending their clinic. The majority of respondents were aware of their child protection policy [122/154 (79%)] and designated child sexual abuse doctor, [125/157 (80%)] in their district. When questioned on previous and current training needs, 134/160 (84%) respondents identified their need for further training in the area of adolescent sexual health and 124/160 (78%) in child sex abuse. The publication Physical Signs of Sexual Abuse in Children, was known to 112/160 (70%) respondents, of whom 58/112 (52%) who answered this question had read the publication. Genitourinary physicians in the UK are aware of the increasing number of children and adolescents accessing their services, and recognize the need to identify those in abusive situations. Written policies dealing with children and adolescents in GUM clinics in the UK are lacking. This needs to be rectified urgently. This survey identifies that further training in the field of child sexual abuse and adolescent sexual health would be welcomed by the respondents. PMID- 11487388 TI - Sexually transmitted infections in Estonia--syndromic management of urethritis in a European country? AB - Sexually transmitted infections (STIs) are considered a major public health problem, globally. In particular, increasing STI rates have been documented throughout eastern Europe and central Asia. The Russian Federation and adjacent countries have, traditionally, managed STIs on an aetiological basis. This approach is expensive in terms of laboratory costs and it may lead to delayed diagnosis and treatment. To overcome the limitations of the aetiological management of STIs, the World Health Organization (WHO) has placed an increased emphasis on integrated care using syndromic management at the primary care level, especially in developing countries. This article reviews the current aetiology of STIs in Estonia, an eastern European country bordering the Baltic Sea and formerly a part of the Soviet Union, with the aim of defining whether infection with Trichomonas vaginalis is common enough to include its management in a syndromic management protocol. The use of syndromic management, in general, is also discussed. PMID- 11487389 TI - An association of disseminated Mycoplasma fermentans in HIV-1 positive patients with non-Hodgkin's lymphoma. AB - We examined the relationship between the haematogenous dissemination of Mycoplasma fermentans and non-Hodgkin's lymphoma (NHL) in 265 HIV-1 positive patients. A polymerase chain reaction (PCR) assay was used to detect M. fermentans in peripheral blood mononuclear cells (PBMCs) from 50 patients enrolled consecutively from an HIV outpatient clinic in 1991 (cohort 1), 56 patients with lower respiratory tract infection who underwent bronchoscopy in 1992 (cohort 2), and 159 patients who were enrolled into a natural history cohort study in 1994 (cohort 3). The incidence of NHL among the patients was determined in 1998. The PBMCs of 29 patients (10.9%) were positive for M. fermentans (8 in cohort 1, 13 in cohort 2 and 8 in cohort 3) and 11 patients (4.2%) developed NHL which was confirmed histologically (3 in cohort 1, 4 in cohort 2 and 4 in cohort 3). We found a statistically significant association between the presence of M. fermentans and the development of NHL in the combined cohort (risk ratio [RR]=6.78 [95% confidence interval (CI) 2.21--20.84], P=0.003 Fisher's exact test [FET]). This association remained significant even after adjustment in a multivariate analysis for CD4 cell count and HIV disease status at the time of M. fermentans testing (RR=7.97 [95% CI=2.16--29.47], P=0.002). PMID- 11487390 TI - Quality of life among urban Canadian HIV/AIDS clinic outpatients. AB - Perceived health-related quality of life (QOL) of 113 adult people living with HIV/AIDS (PHA) attending an outpatient hospital clinic was assessed upon initial registration using the Medical Outcomes Study SF-36. QOL ratings on both physical functioning and psychological well-being aspects of the SF-36 were lower than for the general population, especially among PHA in the sample who were symptomatic or for whom AIDS had developed. Although not compromised in terms of physical functioning, asymptomatic PHA had well-being QOL scores comparable to other chronic medical conditions. In general, demographic, social, and many of the health and immune status variables examined were not significantly related to QOL. Number of symptoms was negatively associated with both physical status and emotional well-being dimensions of QOL. PHA in the asymptomatic stage of HIV infection reported better QOL than symptomatic/AIDS patients on few QOL dimensions. Mean QOL ratings did not significantly differ between AIDS patients and those with symptomatic (non-AIDS) infection. These findings suggest that although becoming symptomatic has a significant impact on a majority of dimensions of QOL, being diagnosed with HIV affects well-being dimensions the most. An emphasis on support for both adjusting to diagnosis and symptom management is as important as viral suppression in maximizing QOL for PHA. PMID- 11487391 TI - Over-the-counter advice for genital problems: the role of the community pharmacist. AB - This year, in the UK, levonorgestrel was approved for sale in pharmacies for emergency contraception. This study assessed, using a postal questionnaire, the ability of community pharmacists to provide advice relating to sexual health, their comfort and training in this area, and their knowledge of local genitourinary medicine (GUM) services. Fifty-four per cent of pharmacists responded. Most (79%) did not know where their nearest GUM department was; only 21% had ever advised a patient to attend a GUM clinic. Twenty-nine per cent said they were not able to broach the possibility of a sexually transmitted infection (STI) with a patient of both sexes. Forty-four per cent had received training related to post-coital contraception. Greater liaison between GUM departments and community pharmacists is suggested as a way of increasing the proportion of patients presenting to a pharmacist who are referred appropriately to a GUM clinic. PMID- 11487392 TI - Seroepidemiology of low and high oncogenic risk types of human papillomavirus in a predominantly male cohort of STD clinic patients. AB - Human papillomaviruses (HPV) infecting the genital tract are associated with warts and anogenital malignancies. Although HPV is a highly prevalent sexually transmitted disease (STD), the majority of research has focused on female cohorts due to gender specific sequelae. Our objective was to measure the epidemiological features and seroprevalences of HPV-6/11 and 16 in a predominantly male group of STD clinic patients. High-risk individuals (n=687), who attended the public STD clinic were administered a behavioural questionnaire and serum tested for antibodies against HPV-6/11 and HPV-16 capsids via capture enzyme-linked immunosorbent assay. Despite the male predominance in this study, women were significantly more likely to have antibodies against both HPV-6/11 and HPV-16. Condom use appeared to be partially protective against HPV-16 seropositivity only. In conclusion, despite exhibiting increased risk behaviour, men were less likely to be HPV seropositive. Additional studies utilizing male cohorts are warranted to further elucidate this phenomenon. PMID- 11487393 TI - HIV seroprevalence and its associations with the other reproductive tract infections in asymptomatic women in Harare, Zimbabwe. AB - The objective was to determine the magnitude of HIV as well as the associations between HIV seropositivity with reproductive tract infections (RTIs) among healthy women in Harare, Zimbabwe. In a cross-sectional study, 393 informed consenting women aged 15--49 years, attending 2 primary healthcare clinics, were interviewed and screened for HIV and RTIs. HIV seroprevalence was 29.3%. Seropositivity was significantly associated with bacterial vaginosis, syphilis, gonorrhoea and/or Chlamydia trachomatis infection, warts and genital ulcers. Risk factors for HIV were absence of lactobacilli in vaginal fluid, vaginal pH >4.5, age >20 years, being unmarried, having had more than one lifetime sexual partner, having used a condom at least once in their lifetime, having experienced an infant mortality, and a partner who was non-monogamous or a frequent traveller. In view of the very high seroprevalence and concomitant RTIs in this population, we recommend promotion of women-controlled prevention methods along with proactive detection and treatment of RTIs. PMID- 11487394 TI - Lipid abnormalities in a healthcare worker receiving HIV prophylaxis. AB - Healthcare workers (HCWs) worldwide risk occupational exposure to HIV and other blood-borne pathogens. Post-exposure prophylaxis (PEP) may decrease the risk of seroconversion after occupational or sexual exposure. Current guidelines recommend immediate PEP with at least 2 drugs following HIV exposure. In high risk exposures, the guidelines recommend the use of a protease inhibitor (PI) as well as 2 reverse transcriptase inhibitors. Protease inhibitors have been associated with dyslipidaemias, other metabolic abnormalities and lipodystrophy syndromes in AIDS patients. We report a case of new transient lipid abnormalities in a HCW receiving PEP after HIV exposure. HIV medications may produce occult metabolic abnormalities in HIV-negative individuals receiving PEP. This risk should be considered during follow-up evaluation for PEP. PMID- 11487396 TI - Do adolescent girls in Bangladesh know about AIDS? PMID- 11487399 TI - Loss and recovery of Arabidopsis-type telomere repeat sequences 5'-(TTTAGGG)(n) 3' in the evolution of a major radiation of flowering plants. AB - Fluorescent in situ hybridization and Southern blotting were used for showing the predominant absence of the Arabidopsis-type telomere repeat sequence (TRS) 5' (TTTAGGG)(n)-3' (the 'typical' telomere) in a monocot clade which comprises up to 6300 species within Asparagales. Initially, two apparently disparate genera that lacked the typical telomere were identified. Here, we used the new angiosperm phylogenetic classification for predicting in which other related families such telomeres might have been lost. Our data revealed that 16 species in 12 families of Asparagales lacked typical telomeres. Phylogenetically, these were clustered in a derived clade, thereby enabling us to predict that the typical telomere was lost, probably as a single evolutionary event, following the divergence of Doryanthaceae ca. 80--90 million years ago. This result illustrates the predictive value of the new phylogeny, as the pattern of species lacking the typical telomere would be considered randomly placed against many previous angiosperm taxonomies. Possible mechanisms by which chromosome end maintenance could have evolved in this group of plants are discussed. Surprisingly, one genus, Ornithogalum (Hyacinthaceae), which is central to the group of plants that have lost the typical telomere, appears to have regained the sequences. The mechanism(s) by which such recovery may have occurred is unknown, but possibilities include horizontal gene transfer and sequence reamplification. PMID- 11487400 TI - A new geographical gradient in vole population dynamics. AB - A new geographical gradient in the dynamics of small rodents is demonstrated by analysing 29 time series of density indices of the common vole (Microtus arvalis) from Poland, the Czech Republic and the Slovak Republic. This gradient extends from more stable northerly populations in coastal Poland to more variable and cyclic populations in the southernmost parts of the Slovak Republic, and is hence a reversal of the Fennoscandian gradient. All studied variables (such as mean density, cycle amplitude, density variability and the coefficients in a second order autoregressive model) exhibit consistent latitudinal variation. Possible underlying factors are discussed. In particular, we suggest that seasonality may be a key element in explaining the observed new gradient. PMID- 11487401 TI - Stabilimenta attract unwelcome predators to orb-webs. AB - Conspicuous behaviour exposes animals to predation; prey-attraction thus often conflicts with antipredator behaviour. The fact that a conspicuous ultraviolet light reflecting silken structure in the orb-webs of certain spider species, known as a stabilimentum, makes the webs obvious to both prey and predators has been used to argue that spiders benefit from building stabilimenta by attracting prey and/or defending against visually hunting predators. Here, we provide experimental evidence that stabilimenta can act as visual signals that attract web-invading spider-eating predators with acute vision to the webs. We also show that the predators can learn to remember a particular type of stabilimentum. Thus, stabilimentum-building spiders risk a high level of predation by attracting visually hunting predators. PMID- 11487402 TI - Dynamic adjustment of parental care in response to perceived paternity. AB - Theories of parental care evolution predict that genetic relatedness will be an important variable in the amount of care a parent provides. However, current inferences of relatedness-based parental investment from studies in humans and birds remain challenged. No study has yet demonstrated parental care adjustment in a manner uncomplicated by life-history correlates or experimental design. We now present a unique test that controls for individual life histories and demonstrates paternity-related dynamic adjustments in parental care. Brood rearing male bluegill sunfish (Lepomis macrochirus) that are cuckolded to a varying degree will either increase or decrease their parental investment in response to changing information on paternity during brood development. Specifically, as parental males detect paternity lost to cuckolders and, hence, a reduction in the value of their brood, they adaptively lower their level of parental care. Conversely, if they detect that their paternity is higher than previously assessed, they adaptively raise their level of parental care. This dynamic adjustment during brood rearing indicates the importance of genetic relatedness in parental investment decisions and provides needed empirical support for theoretical predictions. PMID- 11487403 TI - The role of spatial scale and intraspecific variation in secondary chemistry in host-plant location by Ceutorhynchus assimilis (Coleoptera: Curculionidae). AB - To understand the ecological role of secondary plant compounds in host location by phytophagous insects it is important to consider attraction at different scales in natural populations. The cabbage seed weevil, Ceutorhynchus assimilis, which lays eggs in pods of crucifers where the larvae feed on seed, is attracted to purified extracts of specific glucosinolate-derived volatiles. We considered the possibility that C. assimilis adults are attracted to and preferentially attack patches of plants and/or individual plants producing these volatiles. Using discrete natural populations of Brassica oleracea and Brassica nigra, we found that oviposition was highest in populations of B. oleracea producing high amounts of 3-butenylglucosinolate. No links were found between the other glucosinolates, 2-propenylglucosinolate, 2-hydroxy-3-butenylglucosinolate, 1 indolylmethylglucosinolate or 1-methoxy-3-indolylmethylglucosinolate, and oviposition in B. oleracea. B. nigra, which contains only 2 propenylglucosinolate, was not attacked by C. assimilis. Within populations of B. oleracea, neither oviposition nor the number of seeds eaten was related to the glucosinolate profiles of individual plants. We suggest that C. assimilis adults use 3-butenylglucosinolate-derived volatiles to locate host populations, whereas other cues determine oviposition on individual plants. The consequences of these results for natural selection of glucosinolate phenotypes are discussed. PMID- 11487404 TI - Predators as stressors? Physiological and reproductive consequences of predation risk in tropical stonechats (Saxicola torquata axillaris). AB - Tropical birds usually lay smaller clutches and are less likely to initiate a second brood than their temperate-zone relatives. This reduction in annual fecundity is generally explained as an adaptation either to higher rates of nest predation or to a more limited food supply concurrent with higher adult survival in the tropics. However, the physiological parameters associated with lower annual fecundity in tropical birds have not been well investigated. We compared the annual fecundity, behaviour and a number of physiological parameters of stonechat parents feeding fledged juveniles in territories with and without fiscal shrikes, a predator on adult and fledged birds. Stonechat pairs in territories with shrikes were less likely to initiate a second brood and delayed successive broods compared to pairs in territories without shrikes. After fledging of their young, males showed a greater propensity than females to initiate distraction calls after a human intrusion into their territory and, therefore, invested more in the defence of their young. In territories with shrikes stonechat males had higher initial plasma corticosterone levels and lower body conditions than males in territories without shrikes, suggesting that they were chronically stressed. In contrast, the females from both types of territory had low initial plasma corticosterone levels. We conclude that shrike presence might account for the delay in initiation of a second brood and the reduction in the tendency to initiate a second brood. Whether these effects are mediated by the elevated levels of corticosterone remains to be demonstrated. PMID- 11487405 TI - Bill morphology reflects female independence from male parental help. AB - The study of territorial polygyny in birds has been influential in the development of the theory of social mating systems. Alternative female mating options have been studied within the framework of the polygyny-threshold model and later as the outcome of conflicts of interest between individuals. However, little attention has been given to variations between individual females, and how this affects their mating behaviour. Here, we test the hypothesis that some females are better adapted to raise nestlings without male assistance, and thus to mate polygynously. Specifically, we investigate whether intraspecific variation in female bill morphology is related to mating behaviour. This hypothesis is derived from earlier studies showing that, in both intra- and interspecific comparisons, uniparental care by females is correlated with the catching of larger prey items than when both parents provision the young. Using the polygynous dusky warbler (Phylloscopus fuscatus) as a model species, we found that, in accordance with our prediction, females with deep bills were more likely to mate as a secondary female. Moreover, regardless of mating status, females with deep bills settled in territories with more food and they received less male assistance in feeding their offspring. We argue that females with stronger bills are better adapted to exploit the abundance of large food items in rich territories and thus to raise young on their own. Our results demonstrate the importance of studying variations between individual females, and provide evidence for an extended version of the 'constrained-female hypothesis'. As bill depth is a highly heritable trait, our study strongly suggests that variation in female mating behaviour is not only related to ecological factors and female condition (as shown elsewhere) but also to heritable morphological traits. PMID- 11487406 TI - The developmental and physiological basis of body size evolution in an insect. AB - The evolution of body size is a dominant feature of animal evolution. However, little is known about how the underlying developmental mechanisms that determine size change as body size evolves. Here we report on a case of body size evolution in the tobacco hornworm Manduca sexta that occurred over a period of nearly 30 years. We take advantage of an extensive series of physiological studies performed in the early 1970s that established the parameters that regulate body size in this species and compare their values with those of modern individuals that are descendants of the same colony. We show that three of the five processes that determine adult body size changed during this period, while two remained constant. Changes in these three developmental processes completely account for the observed evolutionary change in body size. PMID- 11487407 TI - Parallel gigantism and complex colonization patterns in the Cape Verde scincid lizards Mabuya and Macroscincus (Reptilia: Scincidae) revealed by mitochondrial DNA sequences. AB - The scincid lizards of the Cape Verde islands comprise the extinct endemic giant Macroscincus coctei and at least five species of Mabuya, one of which, Mabuya vaillanti, also had populations with large body size. Phylogenetic analysis based on DNA sequences derived from the mitochondrial cytochrome b, cytochrome oxidase I and 12S rRNA genes (711, 498 and 378 base pairs (bp), respectively) corroborates morphological evidence that these species constitute a clade and that Macroscincus is unrelated to very large skinks in other areas. The relationships are ((M. vaillanti and Mabuya delalandii) (Mabuya spinalis and Macroscincus coctei (Mabuya fogoensis nicolauensis (Mabuya fogoensis antaoensis and Mabuya stangeri)))). The Cape Verde archipelago was colonized from West Africa, probably in the Late Miocene or Early Pliocene period. The north-eastern islands were probably occupied first, after which the ancestor of M. vaillanti and M. delalandii may have originated on Boavista, the ancestor of the latter species arriving on Santiago or Fogo later. The M. fogoensis--M. stangeri clade colonized the islands of Branco, Razo, Santa Luzia and Sao Vicente from Sao Nicolau and reached Santo Antao after this. Colonization of these northeastern islands was slow, perhaps because the recipient islands had not developed earlier or because colonization cut across the path of the Canary Current and the Northeast Trade Winds, the main dispersing agents in the region. Rapid extension of range into the southwestern islands occurred later in M. spinalis and then in M. vaillanti and M. delalandii. The long apparent delay between the origin of these species and their southwestern dispersal may have been because there were earlier colonizations of the southern islands which excluded later ones until the earlier inhabitants were exterminated by volcanic or climatic events. The evolution of large size in Macroscincus occurred in the northwestern islands and was paralleled in the eastern and southern islands by populations of M. vaillanti. Both cases of size increase in Cape Verde skinks were accompanied by the development of herbivory. PMID- 11487409 TI - Symmetry, sexual dimorphism in facial proportions and male facial attractiveness. AB - Facial symmetry has been proposed as a marker of developmental stability that may be important in human mate choice. Several studies have demonstrated positive relationships between facial symmetry and attractiveness. It was recently proposed that symmetry is not a primary cue to facial attractiveness, as symmetrical faces remain attractive even when presented as half faces (with no cues to symmetry). Facial sexual dimorphisms ('masculinity') have been suggested as a possible cue that may covary with symmetry in men following data on trait size/symmetry relationships in other species. Here, we use real and computer graphic male faces in order to demonstrate that (i) symmetric faces are more attractive, but not reliably more masculine than less symmetric faces and (ii) that symmetric faces possess characteristics that are attractive independent of symmetry, but that these characteristics remain at present undefined. PMID- 11487408 TI - The evolution of armadillos, anteaters and sloths depicted by nuclear and mitochondrial phylogenies: implications for the status of the enigmatic fossil Eurotamandua. AB - The mammalian order Xenarthra (armadillos, anteaters and sloths) is one of the four major clades of placentals, but it remains poorly studied from the molecular phylogenetics perspective. We present here a study encompassing most of the order's diversity in order to establish xenarthrans' intra-ordinal relationships, discuss the evolution of their morphological characters, search for their extant sister group and specify the timing of their radiation with special emphasis on the status of the controversial fossil Eurotamandua. Sequences of three genes (nuclear exon 28 of the Von Willebrand factor and mitochondrial 12S and 16S rRNAs) are compared for eight of the 13 living genera. Phylogenetic analyses confirm the order's monophyly and that of its three major lineages: armadillos (Cingulata), anteaters (Vermilingua) and sloths ('Tardigrada', renamed in 'Folivora'), and our results strongly support the grouping of hairy xenarthrans (anteaters and sloths) into Pilosa. Within placentals, Afrotheria might be the first lineage to branch off, followed by Xenarthra. The morphological adaptative convergence between New World xenarthrans and Old World pangolins is confirmed. Molecular datings place the early emergence of armadillos around the Cretaceous/Tertiary boundary, followed by the divergence between anteaters and sloths in the Early Eocene era. These Tertiary dates contradict the concept of a very ancient origin of modern xenarthran lineages. They also question the placement of the purported fossil anteater (Eurotamandua) from the Middle Eocene period of Europe with the Vermilingua and instead suggest the independent and convergent evolution of this enigmatic taxon. PMID- 11487410 TI - The persistence of Pliocene populations through the Pleistocene climatic cycles: evidence from the phylogeography of an Iberian lizard. AB - Ancient climatic fluctuations have caused changes in the demography and distribution of many species. The genetic differentiation between populations of the same species and of sister species is often attributed largely to the more recent Pleistocene fluctuations. Recent interpretations, which implicate earlier episodes, have proved controversial. We address the timing of genetic divergence in the Iberian lizard Lacerta schreiberi by studying the phylogeography of the cytochrome b sequence. The species has a remarkable morphological uniformity, yet our evidence suggests that earlier events in the Pliocene initiated the main divergence between populations. This interpretation implies that the different populations survived through the Pleistocene in separate localities. This conclusion is robust to different molecular clock calibrations. The persistence of earlier differentiation through the Pleistocene has wide implications for our understanding of Pleistocene refugia in this species and, by extension, to the biogeography of the whole region. PMID- 11487411 TI - Transient post-mating inhibition of behavioural and central nervous responses to sex pheromone in an insect. AB - Mating is costly for both male and female insects and should therefore only occur if it is likely to be successful. Within one scotophase, which is the dark period of the light cycle, male moths can only produce one single spermatophore, which is transferred to the female during mating. Remating within the same scotophase would thus be unsuccessful. We tested the hypothesis that newly mated males of the moth Agrotis ipsilon have developed an energy-saving strategy based on the transient inhibition of their sexual behaviour, thus avoiding unsuccessful remating. Agrotis ipsilon males do not copulate more than once during the same scotophase. Moreover, newly mated males do not respond behaviourally to the female sex pheromone although electroantennograms showed that their peripheral olfactory system is fully functional. However, intracellular recordings of antennal lobe neurons showed that the sensitivity for the synthetic sex pheromone blend decreased as compared with that of unmated males. Both the sexual behaviour and the sensitivity of the antennal lobe neurons were restored when tested during the next scotophase. Our results show a fast, transient neuronal plasticity that 'switches off' the olfactory system, which could prevent males from mating unsuccessfully. PMID- 11487412 TI - Is song-type matching a conventional signal of aggressive intentions? AB - Song-type matching is a singing strategy found in some oscine songbirds with repertoires of song types and at least partial sharing of song types between males. Males reply to the song of a rival male by subsequently singing the same song type. For type matching to serve as an effective long-distance threat signal, it must be backed up by some probability of aggressive approach and impose some type of cost on senders that minimizes the temptation to bluff. Western subspecies of the song sparrow exhibit moderate levels of song-type sharing between adjacent males and sometimes type match in response to playback of song types they possess in their repertoires. Interactive playback experiments were used in order to examine the subsequent behaviour of type-matching birds and to quantify the responses of focal birds to type-matching versus non-matching stimuli. Birds that chose to type match the playback of a shared song type subsequently approached the speaker much more aggressively than birds that did not type match. Moreover, birds approached a type-matching stimulus much more aggressively than a non-matching stimulus. These results and consideration of alternatives suggest that type matching in song sparrows is a conventional signal in which honesty is maintained by a receiver retaliation cost against bluffers. PMID- 11487413 TI - Experimental confirmation of the polygyny threshold model for red-winged blackbirds. AB - The polygyny threshold model assumes that polygynous mating is costly to females and proposes that females pay the cost of polygyny only when compensated by obtaining a superior territory or male. We present, to the authors' knowledge, the first experimental field test to demonstrate that females trade mating status against territory quality as proposed by this hypothesis. Previous work has shown that female red-winged blackbirds (Agelaius phoeniceus) in Ontario prefer settling with unmated males and that this preference is adaptive because polygynous mating status lowers female reproductive success. Other evidence suggests that nesting over water increases the reproductive success of female red winged blackbirds. Here we describe an experiment in which females were given choices between two adjacent territories, one owned by an unmated male without any over-water nesting sites and the other by an already-mated male with over water sites. Females overwhelmingly preferred the already-mated males, demonstrating that superior territory quality can reverse preferences based on mating status and supporting the polygyny threshold model as the explanation for polygyny in this population. PMID- 11487414 TI - From Paris to Stockholm: where does the European Alcohol Action Plan lead to? PMID- 11487415 TI - State-dependent learning for alcohol-dependent people. PMID- 11487416 TI - A paper which must be withdrawn from publication. PMID- 11487417 TI - Conversation with David Penington. PMID- 11487418 TI - Heroin overdose: causes and consequences. AB - Over the past decade fatal opioid overdose has emerged as a major public health issue internationally. This paper examines the risk factors for overdose from a biomedical perspective. While significant risk factors for opioid overdose fatality are well recognized, the mechanism of fatal overdose remains unclear. Losses of tolerance and concomitant use of alcohol and other CNS depressants clearly play a major role in fatality; however, such risk factors do not account for the strong age and gender patterns observed consistently among victims of overdose. There is evidence that systemic disease may be more prevalent in users at greatest risk of overdose. We hypothesize that pulmonary and hepatic dysfunction resulting from such disease may increase susceptibility to both fatal and non-fatal overdose. Sequelae of non-fatal overdose are recognized in the clinical literature but few epidemiological data exist describing the burden of morbidity arising from such sequelae. The potential for overdose to cause persisting morbidity is reviewed. PMID- 11487419 TI - Study of 2708 heroin-related deaths in north-eastern Italy 1985-98 to establish the main causes of death. AB - AIMS: To ascertain the causes of deaths among a very large cohort of heroin injecting drug users (IDUs) who, from 1985 to 1998, attended 36 Public Health Authority Centres for Drug Users (PCDUs) in north-eastern Italy. DESIGN: Retrospective analysis of data, obtained from the Annual Register of each Centre and the Municipal Registry Office of each local health district. SETTING: Thirty six PCDUs in north-eastern Italy and Medical Service for Addictive Disorders of the University of Verona. PARTICIPANTS: All IDUs who had sought medical care at least once in the PCDUs during the study period. FINDINGS: Of 2708 deaths, overdose was found to be the major cause (37%), followed by AIDS (32.5%) and road accidents (9.4%). The percentage of deaths due to AIDS increased steadily from 2.7% in 1985 to 42.2% in 1996, and then decreased to 16.9% in 1998. Deaths due to overdose remained almost constant. The average age of death per year rose from 26 in the mid eighties to 34 in 1998. The mortality rate among IDUs proved much higher compared to the general population of the same age (13-fold, 95% CI, 11.3 14.6). The proportion of all deaths attributable to regular use of illegal opiates in the 15-34 age group in the general population in 1991 was 16%. HIV prevalence was not a significant factor in suicides and deaths by overdose. CONCLUSIONS: The mortality rate was 13 times greater than in the general population. To be female and to have dropped out of any kind of treatment proved an important risk factor for overdose. The fall in deaths from AIDS enhances the problem to prevent and treat HCV infection. Decisions in drug projects, in research and in training should be influenced by the strikingly high percentage of deaths due to drug use. PMID- 11487420 TI - Ecstasy pill testing: harm minimization gone too far? AB - Harm reduction has become the focus of public health initiatives and therapeutic intervention in the management of dependent drug use over the last 20 years. The last decade has seen such approaches being extended to recreational drug use. Most harm reduction initiatives have aimed to inform users about risks and ways of minimizing risk. The concept of providing illicit drug users with quality assessment of their chosen drug is one possible harm reduction intervention that until recently has received little attention. In response to well-publicized 'ecstasy'-related deaths organizations in some European countries and the United States have chosen to provide a 'pill testing service' for users. There are two broad categories of pill testing offered. Simple colour reagent test kits (Marquis Reagent and colour charts) form the most widely used on-site pill testing method. Less frequently, but more accurately, laboratory personnel with access to sophisticated chromatographic equipment (high performance liquid chromatography (HPLC) or gas chromatography-mass spectrometry (GC-MS)) may provide analysis of a pill. Pill testing kits have been advocated as a 'tool to protect yourself against the polluted XTC market'. We refute this line of reasoning. Of the different tests only techniques such as GC-MS can identify satisfactorily the psychoactive constituents present in ecstasy pills. Colour tests based on an interpretation of a colour response in the presence of a drug are, at best, subjective. Pill testing of any description does not guarantee safety, or protect the consumer against individual responses to pills. At best it gives an artificial 'shine of safety' to a group of diverse drugs that remain both illicit and potentially harmful. Other simpler harm reduction mechanisms are likely to be more effective. PMID- 11487421 TI - Attrition prevention with individuals awaiting publicly funded drug treatment. AB - AIMS: To evaluate the effectiveness of a motivational intervention to reduce attrition from a waiting list for substance abusers seeking publicly funded treatment. DESIGN: Randomized clinical trial comparing an "attrition prevention" condition to standard care while awaiting treatment admission. SETTING: A centralized substance abuse assessment and referral center in Seattle, Washington. PARTICIPANTS: Substance abusers (n = 654) eligible for publicly funded drug abuse treatment. MEASUREMENTS: Alcohol and drug use, substance related negative consequences, areas in need of help, perceived need for help, emotional status, readiness to change, reasons for seeking and perceived barriers to entering treatment. FINDINGS: Overall, approximately 70% of clients entered treatment, and of these approximately 70% completed their assigned treatment. Those who entered treatment showed significant reductions in substance use and improved psychosocial function at a short-term 3-month follow-up. However, the attrition prevention intervention had no differential effect on treatment entry, completion or outcome compared to the standard waiting list. Further, there were no differences across therapists on these outcome measures. CONCLUSIONS: A motivational attrition prevention intervention did not enhance treatment entry, completion or outcome among treatment-seeking substance abusers. It is suggested that alternative strategies, such as contingency management and case management, may help facilitate treatment entry for individuals seeking publicly funded treatment. PMID- 11487422 TI - Cue exposure with coping skills training and communication skills training for alcohol dependence: 6- and 12-month outcomes. AB - AIMS: Cue exposure treatment (CET) has shown promise in preliminary studies with alcoholics, and Communication Skills Training (CST) has been found beneficial, especially in intensive treatment programs. The aim of the present study was to investigate the effects of CET and CST in a larger controlled study when both were added to intensive treatment programs. DESIGN AND INTERVENTIONS: A 2 x 2 design investigated the effects of CET with urge coping skills training compared to a meditation-relaxation control, and CST compared to an education control when all were added to intensive treatment programs for alcoholics. SETTING: The sites were the inpatient or partial hospital substance abuse treatment programs at a private psychiatric hospital, a state-funded residential facility and a VA medical center. PARTICIPANTS: Patients diagnosed with alcohol dependence without active psychosis were eligible. MEASUREMENTS: Participants were assessed for quantity and frequency of drinking, alcohol cue-reactivity, responses to high risk simulations and urge specific coping skills. FINDINGS: Of 100 treated patients, 86% provided 6-month and 84% provided 12-month follow-up data. Patients who received either CET or CST had fewer heavy drinking days in the first 6 months than control patients. In the second 6 months, CET continued to result in fewer heavy drinking days among lapsers and interacted with CST to decrease quantity of alcohol consumed. CST resulted in fewer alcohol-related problems reported at 12 months. CET resulted in greater reductions in urge to drink in a measure of simulated high-risk situations. CET also resulted in greater reports of use of coping strategies during the follow-up, and many of the urge-specific strategies taught in CET were associated with reduced drinking. CONCLUSIONS: Both CET and CST continue to show promise as elements of comprehensive alcohol treatment programs. Limitations and directions for future research are discussed. PMID- 11487423 TI - The influence of sensitivity to reward on reactivity to alcohol-related cues. AB - AIMS: To investigate the role of sensitivity to reward in mediating social drinkers' reactivity to alcohol cues. DESIGN: A standard cue-reactivity paradigm was employed. Two groups of social drinkers (heavy and light) were assessed after exposure to the sight, smell and taste of a neutral cue (water) and then an alcohol cue (glass of beer). SETTING: Sessions were conducted in a laboratory based environment. PARTICIPANTS: Twenty heavy (12 males, eight females) and 18 light social drinkers (seven males, 11 females) were recruited; mean age was 23.6 years. MEASUREMENTS: The Card Arranging Reward Responsivity Objective Test (CARROT), assessing behavioural responsiveness to a monetary incentive; urge to drink; positive affect; and the BAS scales, assessing sensitivity to reward. FINDINGS: Heavy drinkers displayed a significant increase in responsivity to rewards (i.e. CARROT) and self-reported urge to drink, but not positive affect, after exposure to alcohol. For the heavy drinkers, heightened sensitivity to reward (i.e. BAS scales) was significantly related to cue-elicited urge to drink and positive affect. CONCLUSION: The results are consistent with a conditioned appetitive motivational model of alcohol use and suggest that Gray's theory of personality may be of some benefit in explaining variation in reactivity responses. PMID- 11487424 TI - The reliability of self-reported age of onset of tobacco, alcohol and illicit drug use. AB - AIMS: To examine the reliability of self-reported age of first substance use experiences among national samples of adult and child respondents. DESIGN: Survey responses from seven waves of the National Longitudinal Survey of Youth (NLSY) were examined. PARTICIPANTS: Adult and child NLSY respondents reporting age of first tobacco, alcohol, marijuana, cocaine and/or crack use during two or more survey interviews. MEASUREMENTS: Four indicators of reliability: intraclass correlation coefficients (ICC), mean and absolute mean differences in reported age and reports consistent within 1 year. FINDINGS: The adjusted mean ICC for all comparisons was 0.69. The adjusted mean difference in self-reported age of first substance use was -0.52 years and the adjusted absolute mean difference was 2.00 years. The adjusted percentage of all comparisons reporting ages consistent within 1 year was 55.28%. More consistent reports were provided by adults, and in response to questions posed over 2 years as opposed to longer time intervals. Respondent answers to questions concerned with first use of marijuana were generally found to be most reliable; questions concerned with first use of crack were least reliable and reports of tobacco, alcohol and cocaine were intermediate. Logistic regression analyses also identified age, race, gender, education and poverty status as predictors of consistent reporting. CONCLUSIONS: Self-reports of age of first substance use experiences, as currently collected via survey questionnaires, are of sufficient reliability for most current epidemiological applications. For inquiries where age of substance use onset is itself a research focus, however, researchers should invest additional effort in improving the reliability of measurement. PMID- 11487425 TI - Pre-pregnancy drinking: how drink size affects risk assessment. AB - AIMS: We considered the role of drink size in determining average daily consumption among groups at risk for Fetal Alcohol Syndrome. DESIGN: In-person hour-long interviews gathered cross-sectional retrospective data about drinking before an index pregnancy. SETTING: Subjects were recruited at public clinics in Los Angeles and the San Francisco Bay area. To reach those not necessarily seeking prenatal care, community outreach in the same urban areas was undertaken. PARTICIPANTS: Three hundred and twenty-one pregnant women were interviewed: 102 Native Americans, 185 African Americans, and 34 Caucasians. MEASUREMENTS: Volume of drinking prior to pregnancy was assessed using the graduated frequency series, which asks respondents to specify their drinking in terms of standard drinks. Using vessel models and photographs, respondent-defined drink sizes were then determined, and volume was recalculated accordingly. FINDINGS: For most beverages, the difference in milliliters between self-selected drink size and a standard size drink was significant, with the mean self-selected drink sizes ranging from 49% above the standard size (for beer) to 307% above the standard size (for spirits). For women whose pre-pregnancy average daily volume (ADV) was at the risk level of > or = 1 standard drink per day, ADV increased from four to almost 10 standard drinks per day when self-defined drink sizes were instead considered. Similarly, for women having three or more standard drinks a day, their daily dose of ethanol increased from 57 g to 153 g of ethanol per day. CONCLUSIONS: If risk levels have been based on underestimates that assume women with alcohol-affected infants had standard drink sizes, then true risk levels may be higher than previously thought. Related, risk drinkers presenting at prenatal clinics may be missed if screening protocols do not ask about drink size. PMID- 11487426 TI - Needle fixation--what is the prevalence? PMID- 11487446 TI - The use of statins in acute coronary syndromes: the mechanisms behind the outcomes. AB - Lipid-lowering drugs, in particular statin treatments, have been shown to reduce the incidence of initial and recurrent coronary heart disease (CHD) events within several years of initiating therapy. This effect can be clinically detected within the first 1 to 2 years in randomized trials. Recent observational and clinical trial data suggest that lipid-lowering therapy initiated at the time of an acute coronary event can reduce recurrent events, and possibly all-cause mortality, in a much shorter period of time. The possible mechanisms by which this benefit occurs include the effect of reduced lipoprotein levels, as well as an independent effect of statins on endothelial function. Statins improve endothelial-dependent flow-mediated vasodilation by increasing the bioavailability of nitric oxide. They stabilize the plaque by modulating the inflammatory response within the vessel wall. They also decrease clot formation by decreasing the adherence of platelets to the ruptured plaque and by acting on the extrinsic coagulation cascade pathway. This review examines these effects of statins and lipoproteins on vascular function, as well as the clinical evidence supporting early treatment in acute coronary syndromes. PMID- 11487447 TI - New developments in the prevention of atherosclerosis in patients with low high density lipoprotein cholesterol. AB - Epidemiologic studies have established that a low concentration of plasma high density lipoprotein (HDL) cholesterol is an independent risk factor for coronary heart disease (CHD). In the United States, a low HDL cholesterol concentration is the most prevalent lipid abnormality observed in men with known CHD. Despite this fact, surprisingly few clinical trials have been designed to investigate the effects of pharmacologic agents on HDL cholesterol-raising and CHD risk in large populations, perhaps due, in part, to the lack of available drugs having significant HDL cholesterol-raising potential. The purpose of this report is to review recent primary and secondary prevention trials that have explored the relationships between drug therapy, HDL cholesterol concentration, and CHD events or progression. Emphasis will be placed on the results of the Veterans Affairs High-Density Lipoprotein Trial, a study that was specifically designed to test the hypothesis that HDL cholesterol-raising with gemfibrozil would reduce CHD morbidity and mortality in patients with CHD whose primary lipid abnormality was a low level of HDL cholesterol. PMID- 11487448 TI - Combination lipid-altering therapy: an emerging treatment paradigm for the 21st century. AB - For the care of an expanding segment of the US population with multiple coronary risk factors, combination lipid-altering therapy is emerging as a treatment imperative. The most recent National Cholesterol Education Program's consensus guidelines emphasize long-term global coronary heart disease (CHD) risk status, designate patients with CHD risk equivalents (eg, diabetes, peripheral arterial disease, 20% or more 10-year absolute CHD risk) for aggressive lipid-altering therapy, and deem the metabolic syndrome (eg, obesity, insulin resistance, hypertension, elevated triglycerides, low levels of high-density lipoprotein cholesterol, small dense low-density lipoprotein particles) as a secondary target for intervention. With the advancing age of the US population and the high prevalence of diabetes, the metabolic syndrome, and CHD, increasing numbers of patients will require a more balanced metabolic attack attainable only through combination lipid-altering regimens. Many of these patients, as well as persons at heightened risk for cardiovascular disease because of a range of heritable conditions (eg, familial hypercholesterolemia, familial combined hyperlipidemia), will undoubtedly require binary or ternary regimens involving statins in concert with niacin, fibric-acid derivatives, or bile acid resins. Such approaches enable the clinician to exploit the complementary effects of these agents, allowing them to be administered at low, optimally tolerable doses that are consistent with superior efficacy and a lower risk of adverse events as compared with escalating doses of monotherapy. PMID- 11487449 TI - Hyperglycemia and cardiovascular disease. AB - Diabetes is associated with many microvascular and macrovascular complications. Hyperglycemia plays a pivotal role in the development of microvascular complications, but the actual effect of hyperglycemia on the development and progression of macrovascular complications remains unclear and even somewhat controversial, particularly in type 2 diabetes. Macrovascular complications are increased in individuals with type 2 diabetes long before there is significant hyperglycemia, and in many, but not all, studies a clear association of glucose elevations and macrovascular complications cannot be shown. The complicated nature of metabolic abnormalities in type 2 diabetes and the relative role of these associated conditions in the development of macrovascular disease make definitive conclusions somewhat difficult. In spite of these considerations, there are certain aspects of hyperglycemia associated with macrovascular disease, particularly elevations of postprandial glucoses, and a number of basic mechanisms to explain these associations that could lead to the development of cardiovascular disease. Some of these basic abnormalities include activation of the sorbital pathway, oxidative stress, advanced glycation endproducts (AGE), and AGE precursors. These changes can result in many abnormalities, such as endothelial dysfunction, alteration of protein function, increased cytokine production, and glycosylation of structural proteins. These considerations suggest that hyperglycemia may play an important, but as yet not clearly defined, role in clinical macrovascular disease. Pursuant to this, several major multisite studies are currently underway to clarify the role of hyperglycemia in cardiovascular disease in type 2 diabetes. PMID- 11487450 TI - Did the antioxidant trials fail to validate the oxidation hypothesis? AB - Most clinical trials on antioxidants using vitamin E or beta-carotene have failed to note any significant change in cardiovascular endpoints. The results of these studies have been interpreted as a setback for the oxidation hypothesis. An analysis of the hypothesis and the trials, however, points out major misconceptions about the hypothesis and unjustified outcome expectations. Wrong selection of patient population, endpoints that are incompatible with the hypothesis, poor choice of antioxidants, and lack of inclusion of biochemical markers of oxidative stress and markers of vascular response are some of the contributors to the "failure" of these trials. PMID- 11487451 TI - Hormone replacement therapy for prevention of coronary heart disease: current evidence. AB - Postmenopausal estrogen replacement, with or without progestin therapy, has a generally favorable impact on lipids, improves endothelial function, and has anti inflammatory and antioxidant effects. These properties should favorably impact coronary risk; indeed, epidemiologic studies have consistently associated hormone replacement therapy with reduced coronary risk. Nonetheless, the Heart & Estrogen/progestin Replacement Study (HERS), a randomized, placebo-controlled, secondary prevention trial of conjugated estrogen with progestin, found no overall reduction in coronary events among women assigned to active hormone treatment. This review explores the role of estrogen replacement among interventions intended to prevent coronary heart disease in the post-HERS era. PMID- 11487452 TI - Platelet-endothelial interactions in atherosclerosis. AB - The pathogenesis of atherosclerosis, the leading cause of morbidity and mortality in the United States, is multifactorial. Many factors that have been shown to influence the development of atherosclerosis also affect the function of the endothelium through soluble or cell-cell interactions. Among these, interactions between platelets and endothelial cells have only recently begun to receive systematic study. This article reviews recent evidence showing how the interaction between platelets and endothelial cells may play a important role in the pathogenesis of atherosclerosis, suggesting an underappreciated potential locus for pharmacologic intervention. PMID- 11487453 TI - New developments in atherosclerosis imaging: electron beam tomography. AB - Recently published data have greatly expanded the applicability of electron beam tomography (EBT) and electron beam angiography (EBA). The prognostic power of coronary artery calcium scoring for cardiac events and associated obstructive disease far surpasses that of conventional risk factors, and will increasingly be incorporated into risk assessment and treatment guidelines. EBT leads to the identification of treatable, non-low-density lipoprotein metabolic disorders that contribute to plaque formation, and to appropriate selection of stress test candidates. Change in calcium score can be used to assess the efficacy of lipid therapy and will serve as a surrogate marker in drug studies. EBA provides effective, noninvasive visualization of native coronary arteries and bypass grafts. PMID- 11487454 TI - Shingles (Herpes Zoster) and Post-herpetic Neuralgia. AB - During childhood chickenpox, varicella-zoster virus becomes latent in neurons of the dorsal root or trigeminal ganglia. Shingles results years to decades later from a breakdown of viral latency within a ganglion and subsequent virus spread to the skin producing a unilateral dermatomal vesicular rash accompanied by segmental pain. Treatment with famciclovir, valacyclovir, and high dose acyclovir is beneficial if started within the first 3 days of the rash. All three drugs can be given orally, are equally effective, shorten the duration of viral shedding and time to healing of the rash by 1 to 2 days, and lessen the intensity and duration of the acute neuritic pain. Famciclovir and valacyclovir have more convenient dosing schedules (three times daily) compared to acyclovir (five times daily). Mild cases of shingles in younger healthy individuals often do not require any antiviral treatment. Pain in shingles may have burning, lancinating, or allodynic qualities, ranges in intensity from mild to unbearable, and lasts 2 to 8 weeks. Pain treatment varies on the type and intensity of pain experienced. In a few patients, post-herpetic neuralgia develops and the dermatomal pain persists for months to years. Effective treatment of post-herpetic pain is often difficult. PMID- 11487455 TI - Cryptococcal Meningitis. AB - Cryptococcal meningitis, often seen in immunocompromised hosts, is also a disease of the immune-competent individual. The diagnosis of cryptococcal meningitis requires a lumbar puncture with measurement of the opening pressure, standard laboratory assessment including cell count, protein and glucose, fungal culture, and cryptococcal polysaccharide antigen. Serum cryptococcal antigen is of great diagnostic value in individuals infected with HIV. Hospital admission for initial therapy with amphotericin B desoxycholate is required. Adjuvant oral therapy with flucytosine for the first 2 weeks of therapy is strongly recommended. If flucytosine is not well tolerated, it may be discontinued with close monitoring and follow-up of cerebrospinal fluid (CSF) response to therapy. Good hydration and appropriate premedication concomitant to the use of amphotericin B are useful interventions preventing side effects. Occasionally, amphotericin B needs to be discontinued due to intolerance or side effects. After CSF sterilization is completed, therapy can be switched to oral fluconazole. Fluconazole is well absorbed orally. There is rarely a need to give intravenous fluconazole. PMID- 11487456 TI - Tuberculosis Meningitis. AB - Treatment of tuberculous meningitis should begin with an anti-tuberculous regimen of at least three drugs: isoniazid, pyrazinamide, and rifampin. Early in the course of therapy, ethambutol or streptomycin may be of some added benefit. If the local incidence of drug resistance to Mycobacterium tuberculosis is greater than 4%, or is unknown, then a fourth drug (ethambutol or streptomycin) should be added. If the patient is from an area with organisms resistant to multiple drugs, or is likely to be infected with a multiply resistant organism for any reason, then the patient should be on enough drugs to insure that at least two active anti-tubercular drugs are included in the therapy. An expert should be consulted Length of therapy is not standardized. For sensitive organisms, a regimen of three drugs daily for 2 months, followed by two-drug therapy (isoniazid and rifampin) has been recommended. The American Thoracic Society (ATS) and the Centers for Disease Control (CDC) have recommended a minimum of 12 months of therapy for tuberculous meningitis. If cultures remain positive for extended periods, or signs or symptoms respond slowly, therapy should be extended to 18 months. Patients with HIV also may need longer courses of therapy. The severity of tuberculous meningitis can be classified based on a system devised by the British Medical Research Council. Stage I patients are fully conscious, rational, and do not have neurologic signs. Stage II patients are confused or have neurologic signs such as cranial nerve palsy or hemiparesis. Stage III patients are comatose or stuporous with more severe neurologic signs. Corticosteroids are recommended if the patient is mentally confused, has neurologic signs, or is comatose (Stages II and III). In patients with moderate disease (Stage II), corticosteroids appear to improve neurologic sequelae and survival. Dexamethasone 6 to 12 mg per day and prednisone 60 to 80 mg per day tapered over 4 to 8 weeks has been used. Symptoms of central nervous system (CNS) inflammation may recur if the corticosteroid taper is implemented too soon or too fast. Steroids and diuretics such as furosemide and acetazolamide are sometimes used to treat hydrocephalus. Ventriculoperitoneal or ventriculoatrial shunting may be required to relieve signs and symptoms of hydrocephalus. PMID- 11487457 TI - St. Louis Encephalitis and West Nile Virus Encephalitis. AB - St. Louis encephalitis virus is a major cause of epidemic arboviral encephalitis in the US. Transmitted by a mosquito vector, this virus is an annual public health concern during the late summer and early fall in much of the midwest and southeast. The characteristic epidemic features of this viral encephalitis coupled with public health surveillance and vector monitoring programs have made the diagnosis readily accessible during the past decade. Recently, however, the arboviral landscape in the US changed dramatically with the emergence and persistence of West Nile virus and associated human neurologic illness in New York and the Northeast. In its New York presentation, West Nile virus encephalitis exhibited clinical and laboratory similarities to St. Louis encephalitis. Not surprisingly, this led to initial confusion in establishing the diagnosis. In anticipation of the potential geographic spread of West Nile virus beyond the northeastern US, neurologists must now consider West Nile virus along with St. Louis encephalitis when diagnosing patients with suspected epidemic mosquito-borne viral encephalitis or meningoencephalitis. Although no specific antiviral agents are yet available, patients will benefit from close monitoring during the initial phase of illness, supportive critical care, and appropriate rehabilitation. PMID- 11487458 TI - Increased Intracerebral Pressure Following Stroke. AB - Increased intracerebral pressure with lethal herniation still accounts for high mortality rates in patients with massive strokes. Patients that are likely to develop increased intracranial pressure can often be identified within the first few hours after stroke onset. Although medical management seems to fail in most of these patients, early hemicraniectomy and induced moderate hypothermia (32;C to 33;C) represent two novel therapeutic approaches to improve neurologic outcomes and decrease mortality rates. PMID- 11487459 TI - Antithrombotic Secondary Prevention After Stroke. AB - In patients with transient ischemic attack (TIA) or ischemic stroke of noncardiac origin, antiplatelet drugs are able to decrease the risk of stroke by 11% to 15%, and decrease the risk of stroke, myocardial infarction (MI), and vascular death by 15% to 22%. Aspirin leads to a moderate but significant reduction of stroke, MI, and vascular death in patients with TIA and ischemic stroke. Low doses are as effective as high doses, but are better tolerated in term of gastrointestinal side effects. The recommended aspirin dose, therefore, is between 50 and 325 mg. Bleeding complications are not dose-dependent, and also occur with the lowest doses. The combination of aspirin (25 mg twice daily) with slow release dipyridamole (200 mg twice daily) is superior compared with aspirin alone for stroke prevention. Ticlopidine is effective in secondary stroke prevention in patients with TIA and stroke. For some endpoints, it is superior to aspirin. Due to its side effect profile (neutropenia, thrombotic thrombocytopenic purpura ), ticlopidine should be given to patients who are intolerant of aspirin. Prospective trials have not indicated whether ticlopidine is suggested for patients who have recurrent cerebrovascular events while on aspirin. Clopidogrel has a better safety profile than ticlopidine. Although not investigated in patients with TIA, clopidogrel should also be effective in these patients assuming the same pathophysiology than in patients with stroke. Clopidogrel is second-line treatment in patients intolerant for aspirin, and first-line treatment for patients with stroke and peripheral arterial disease or MI. A frequent clinical problem is patients who are already on aspirin because of coronary heart disease or a prior cerebral ischemic event, and then suffer a first or recurrent TIA or stroke. No single clinical trial has investigated this problem. Therefore, recommendations are not evidence-based. Possible strategies include the following: continue aspirin, add dipyridamole, add clopidogrel, switch to ticlopidine or clopidogrel, or switch to anticoagulation with an International Normalized Ratio (INR) of 2.0 to 3.0. The combination of low-dose warfarin and aspirin was never studied in the secondary prevention of stroke. In patients with a cardiac source of embolism, anticoagulation is recommended with an INR of 2.0 to 3.0. At the present time, anticoagulation with an INR between 3.0 and 4.5 can not be recommended for patients with noncardiac TIA or stroke. Anticoagulation with an INR between 3.0 and 4.5 carries a high bleeding risk. Whether anticoagulation with lower INR is safe and effective is not yet known. Treatment of vascular risk factors should also be performed in secondary stroke prevention. PMID- 11487460 TI - Spontaneous Arterial Dissection. AB - There is no controlled study for the best treatment or management of cervico cerebral artery dissection (CAD). Rationale initial empiric treatment in acute CAD to prevent secondary embolism is partial thromboplastin time (PTT)-guided anticoagulation by intravenous heparin followed by anticoagulation with warfarin. Carotid surgery for treatment of CAD is not recommended anymore with the possible exception of persisting severe stenosis of the proximal internal carotid artery (ICA). There could be use of carotid angioplasty by balloon dilatation and stenting in selected cases of severe cerebral hemodynamic impairment by bilateral CAD. Duration of secondary prophylaxis by anticoagulation is best guided by Doppler sonography follow-up, and should be continued until normalization of blood flow or until at least 1 year after the vessel is occluded. There is no evidence that pseudoaneurysms increase the risk for embolic complication, and there is no evidence for surgery or continuation of anticoagulation in patients with pseudoaneurysms. Caution should be recommended for exercises that involve excessive head movements (eg, bungee jumping, trampoline jumping, and chiropractic maneuvers). The patient should be informed that recurrent rate is low in nonfamilial cases. Doppler sonography is a low-cost and high-sensitivity method for patients at risk. PMID- 11487461 TI - Acute and long-term neurodevelopmental outcomes in children following bone marrow transplantation. AB - Bone marrow transplantation offers a potential cure for a number of childhood cancers, sickle cell anemia, and stabilization of a deteriorating and debilitating process in a number of metabolic disorders and leukodystrophies. Depending upon the disease, treatment prior to BMT, and natural history of the disease, BMT may increase the risk of neuropsychological toxicity for children undergoing BMT, or may actually improve their long-term neurodevelopmental outlook. The role of factors such as pre-BMT therapy, age at time of treatment, presence or absence of total body irradiation, and toxicities associated with GVHD are presented for consideration. A developmental model for understanding the emergence of neurocognitive effects of BMT is reviewed, and strategies for intervention are considered. PMID- 11487462 TI - Immune reconstitution in pediatric stem-cell transplantation. AB - Hematopoietic stem-cell transplantation (HSCT) has successfully been used to cure many pediatric disorders. However, the immunologic alterations associated with transplantation result in profound immunodeficiencies in the transplant recipient, resulting in significant infectious morbidity and mortality. The precarious process of immune reconstitution in the transplant recipient is neither instantaneous nor complete, but influenced by multiple factors such as graft-versus-host disease (GVHD), conditioning regimen, patient age and underlying disease. Studies in pediatric HSCT have revealed unique attributes of immune recovery in pediatric transplant recipients. Future studies addressing these findings are needed to complement novel immunotherapies emerging from the field of transplant immunology. PMID- 11487464 TI - Importance of the regulation of nuclear receptor degradation. AB - Nuclear hormone receptors (NHRs) represent a superfamily of structurally related ligand-activated transcription factors, which regulate diverse biological activities like growth, development, and homeostasis. Recently, it has been demonstrated that certain members of the NHR superfamily are degraded through the ubiquitin-proteasome pathway in a ligand-dependent manner. Though the signal for the down-regulation via the ubiquitin-proteasome pathway is not yet known, phosphorylation at specific amino acid residues or coactivator binding to receptors could lead to their degradation by the 26S proteasome. Activation and degradation seems to be an engineered cyclic mechanism, which provides tight control over diverse cellular processes. The degradation process involves extensive loss of proteins and requires expenditure of cellular ATP. That seems to be inevitable for a more important aim, that is efficient and appropriate regulation of transcription. Down-regulation of receptors would lead to an attenuated transcriptional response because the number of receptor molecules available to activate transcription would decrease over time. One of the obvious reasons for down-regulating NHRs thus seems to be to prevent the cell from overstimulation by the hormones or other activating signals. Nuclear receptor turnover may also reset the transcriptional apparatus in preparation for a subsequent response. Since inhibition of the ubiquitin-proteasome degradation pathway disturbs the transcriptional activitity of some of the nuclear receptors such as estrogen (ER) and progesterone (PR) receptors, it is also possible that the degradation of NHRs may enable recycling of components of receptor-cofactor complexes and general transcriptional machinary. Understanding the mechanism of nuclear hormone receptor degradation and its relation to transcription may lead to novel insights of therapuetic intervention. PMID- 11487463 TI - Duality in bromodomain-containing protein complexes. AB - Proteins that contain a motif called a bromodomain are implicated in both transcriptional activation and repression. The bromodomain of p/CAF, the only solution structure of a bromodomain that has been solved to date, reveals that the motif binds N-acetyl-lysine groups, presumably to anchor enzymatic functions to histones and by extension to chromatin. The enzymatic activities can either be encoded within the same polypeptide as the bromodomain motif, or associated with a multiprotein complex. Thus, a wide variety of chromatin-directed functions, including but not limited to phosphorylation, acetylation, methylation, transcriptional co-activation or recruitment, characterize the complexes that contain bromodomain motifs. Their versatility and ubiquity ensures diverse, rapid and flexible transcriptional responses. PMID- 11487465 TI - Acetyllysine-binding and function of bromodomain-containing proteins in chromatin. AB - Acetylated histones are generally associated with active chromatin. The bromodomain has recently been identified as a protein module capable of binding to acetylated lysine residues, and hence is able to mediate the recruitment of factors to acetylated chromatin. Functional studies of bromodomain-containing proteins indicate how this domain contributes to the activity of a number of nuclear factors including histone acetyltransferases and chromatin remodelling complexes. Here, we review the characteristics of acetyllysine-binding by bromodomains, discuss associated domains found in these proteins, and address the function of the bromodomain in the context of chromatin. Finally, the modulation of bromodomain binding by neighbouring post-translational modifications within histone tails might provide a mechanism through which combinations of covalent marks could exert control on chromatin function. PMID- 11487466 TI - The bromodomain: a chromatin browser? AB - Reversible modification of histone tails is a regulatory step in chromatin remodeling. The N-terminal tails of histones are signaling platforms that carry amino acid residues for post-translational modification and contribute to chromosomal higher order structure. These modifications are performed by a number of chromatin modulators such as histone (h) acetyltransferase, h-deacetylase, h methyltransferase and h-kinase. Large numbers of these enzymes as well as other chromatin-associated proteins share the bromodomain, a signature protein motif. Structural studies reveal not only wide structural conservation of bromodomains but also envision a possible role of this domain in the recognition of specific modified residues in the histone tails. The widespread presence of bromodomains in leukemogenic and cancer genes has provided a fundamental tool for studies of the role of epigenetic and chromatin remodeling in malignant diseases. PMID- 11487468 TI - You bet-cha: a novel family of transcriptional regulators. AB - The BET proteins are a novel class of transcriptional regulators whose members can be found in animals, plants and fungi. Founding members are Human RING3, Drosophila Fsh and yeast Bdf1p. BET proteins are distinguished by an N-terminal bromodomain or bromodomains and an ET domain. As predicted by the presence of the bromodomain(s), these proteins have been found to be associated with chromatin. The poorly characterized ET domain functions as a protein-protein interaction motif and may be part of a serine-kinase activity. Other regions ("modular domains"), which are conserved only in certain BET proteins, are likely to provide sub-family specific functions. Genetic, biochemical and molecular techniques have implicated BET proteins in functions as diverse as meiosis, cell cycle control and homeosis. The data suggest that BET proteins may modulate chromatin structure and affect transcription via a sequence-independent mechanism. This review will attempt to summarize current research on BET proteins and envision where future research is likely to lead. PMID- 11487467 TI - Cell adhesion in the preimplantation mammalian embryo and its role in trophectoderm differentiation and blastocyst morphogenesis. AB - Cell adhesion plays a critical role in the differentiation of the trophectoderm epithelium and the morphogenesis of the blastocyst. In the mouse embryo, E cadherin mediated adhesion initiates at compaction at the 8-cell stage, regulated post-translationally via protein kinase C and other signalling molecules. E cadherin adhesion organises epithelial polarisation of blastomeres at compaction. Subsequently, the proteins of the epithelial tight junction are expressed and assemble at the apicolateral contact region between outer blastomeres in three phases, culminating at the 32-cell stage when blastocoel cavitation begins. Cell adhesion events also coordinate the cellular allocation and spatial segregation of the inner cell mass (ICM) of the blastocyst, and the maintenance of epithelial (trophectoderm) and non-epithelial (ICM) phenotypes during early morphogenesis. PMID- 11487469 TI - Human MHC class III and IV genes and disease associations. AB - The major histocompatibility complex (MHC) was initially defined as the genetic locus encoding the Class I and Class II highly polymorphic cell surface antigens that are now known to present antigen to matched sets of T cell receptors. Genes for several diverse complement components, specifically Bf, C2, and C4 were found between the Class I and II genes, in a region later dubbed Class III. More recently, several genes have been described that are encoded in the telomeric end of the Class III region and that appear to be involved in both global and specific inflammatory responses. Due to this commonality of function this gene rich region was dubbed Class IV, and includes the TNF family, AIF1, and HSP70. The genes of the Class III and Class IV regions are sufficiently divergent in sequence and structure so that clustering is not explicable in terms of gene duplication or divergence. We present some of the newer pertinent information and puzzling features of the genes embraced in the Class IV region and discuss possible roles in specific autoimmune diseases linked to this region. PMID- 11487470 TI - Utilizing biomimetic oligopeptides to probe fibronectin-integrin binding and signaling in regulating macrophage function in vitro and in vivo. AB - Biomimetic oligopeptides were employed to elucidate the molecular mechanisms of fibronectin-integrin interaction in regulating macrophage function. Oligopeptides were designed based on of the functional structure of fibronectin and grafted onto a polymer network containing polyethyleneglycols. Macrophage adhesion was independent of the peptide identity that contained sequence RGD, PHSRN, PRRARV, or combinations thereof in an integrin-dependent fashion in vitro. However, integrin-dependent foreign body giant cell (FBGC) formation in vitro was highly dependent on both RGD and PHSRN in a single peptide formulation and with a specific orientation. In vivo results showed that peptide identity played a minimal role in modulating the host inflammatory response and adherent macrophage density. RGD-containing peptides mediated a rapid FBGC formation by 4 days of implantation by significantly increasing both the number of macrophages that participate in the cell fusion process and the rate of cell fusion. Both RGD and PHSRN domains were important in mediating FBGC formation at later implantation periods. In vitro intracellular signaling studies revealed that the requirement of protein tyrosine kinase and serine/threonine kinase activation and cross-talk compensation for macrophage adhesion dynamically varied with surfaces and culture time. Protein kinase C-dependent adhesion was related to RGD and PHSRN sequences, and to the sequence orientation thereof in a form of GGGRGDGGGGGGPHSRNG. Furthermore, we observed a multiple effect of the mitogen-activated protein kinase/extracellular-signal-regulated kinase signaling factor in mediating macrophage adhesion, which depended on the method of ligand immobilization. These findings represent a mechanistic correlation between the role of substrates and protein functional architectures in ligand-receptor recognition and post-ligation signaling events that control cellular behavior in vitro and in vivo. PMID- 11487472 TI - The structure and functions of human lysophosphatidic acid acyltransferases. AB - Lysophosphatidic acid (LPA) and phosphatidic acid (PA) are two phospholipids involved in signal transduction and in lipid biosynthesis in cells. LPA acyltransferase (LPAAT), also known as 1-acyl sn-glycerol-3-phosphate acyltransferase (1-AGPAT) (EC 2.3.1.51), catalyzes the conversion of LPA to PA. Two human isoforms of LPAAT, designated as LPAAT-alpha (AGPAT1) and LPAAT-beta (AGPAT2), have been extensively characterized. These two proteins contain extensive sequence similarities to microbial, plant and animal LPAAT sequences. LPAAT-alpha mRNA is uniformly expressed throughout most tissues with the highest level found in skeletal muscle; whereas LPAAT-beta is differentially expressed, with the highest level found in heart and liver, and negligible level in brain and placenta. The LPAAT-alpha gene is located on chromosome 6p21.3, an area within the class III region of the major hiscompatibility complex (MHC) and the LPAAT-beta gene is mapped to chromosome 9q34.3. Enhanced transcription of LPAAT beta is suggested for neoplasm of the female genital tract. Additionally, ectopic LPAAT expression in certain cytokine-responsive cell lines can effect amplification of cellular signaling processes, such as those leading to enhancement of synthesis of tumor necrosis factor-alpha and interleukin-6 from cells following stimulation with interleukin-1beta; this suggests that the LPAAT genes represent candidates for affecting the development of certain cancers or inflammation-associated diseases. PMID- 11487471 TI - Reactive carbonyl formation by oxidative and non-oxidative pathways. AB - The spectrophotometric protein carbonyl assay is used as an indicator of protein damage by free radical reactions in vitro and in a variety of pathologies. We investigated model proteins and a variety of oxidative and non-oxidative reactions, as well as what effects hemoglobin, myoglobin, and cytochrome c might have on levels of protein carbonyls. We show that oxidative as well as non oxidative mechanisms introduce carbonyl groups into proteins, providing a moiety for quantification with 2,4-dinitrophenylhydrazine (DNPH). Bovine serum albumin exposed to oxidative scenarios, such as hypochlorous acid, peroxynitrite, and metal-catalyzed oxidation exhibited variable, but increased levels of carbonyls. Other non-oxidative modification systems, in which proteins are incubated with various aldehydes, such as malondialdehyde, acrolein, glycolaldehyde, and glyoxal also generated significant amounts of carbonyls. Furthermore, purified myoglobin, hemoglobin, and cytochrome c show high absorbance at the same wavelengths as DNPH. The high levels observed are due to the innate absorbance of hemoglobin, myoglobin, and cytochrome c near the assay spectra of DNPH. These studies show that carbonyl content could be due to oxidative as well as non-oxidative mechanisms and that heme-containing compounds may effect carbonyl quantification. PMID- 11487473 TI - Role of major histocompatibility complex class III genes in recurrent spontaneous abortions. AB - Increased parental Human Leukocyte Antigen (HLA) sharing has been repeatedly reported in recurrent spontaneous abortions (RSA). Parental HLA sharing increases the chance of feto-maternal histocompatibility and potentially affects maternal allo-recognition of the fetus. However, strong linkage disequilibrium across the whole Major Histocompatibility Complex (MHC) region makes it difficult to interpret parental HLA sharing conclusively. It is not known whether the shared HLA gene as such or an unknown gene(s) in linkage disequilibrium or a combination of several loci are causing the disease. Interestingly, in mouse and rat MHC linked, recessive genes are known to control the reproduction, development and growth of the fetus. Human analogs have not been identified. Compared to HLA genes, MHC Class III has been studied much less in RSA. However, there are some observations of an increased number of unexpressed complement C4 alleles in RSA spouses. Complement C4 genes are located in a chromosomal region characterized by extremely high gene density and frequent gene rearrangements. C4 "null" alleles can act as markers of gene rearrangements in Class III unfavorable for pregnancy outcome. Many of the novel genes located in this region by sequencing serve as new candidates for RSA, since they have housekeeping functions and some of them are highly expressed in human reproductive organs. PMID- 11487474 TI - Nonmyeloablative hematopoietic stem cell transplantation: transplantation for the 21st century. AB - Conventional approaches to allogeneic stem cell transplantation have used toxic high-dose conditioning therapy to achieve allogeneic engraftment and control of underlying disease. For engraftment purposes, preclinical studies and clinical observations have shown that conditioning regimens can be markedly reduced in intensity, resulting in reduced treatment toxicities. Preclinical canine studies demonstrated that the use of potent pre- and postgrafting immunosuppression allows for reduction in conditioning regimens while facilitating development of stable mixed chimerism. If attenuated conditioning regimens can be successfully translated to human stem cell transplantation, an improved safety profile will allow potentially curative treatment to a more representative patient profile not currently offered such therapy. Mixed chimerism could prove curative of disease phenotype of various nonmalignant disturbances of the hematopoietic and immune systems. For patients with hematopoietic malignancy, spontaneous conversion to full donor hematopoeisis after stem cell transplant may prove curative by virtue of graft versus host reactions directed against the malignancy, however infusion of additional donor lymphocytes may be needed to treat persistent disease. PMID- 11487475 TI - Genetics of human complement component C4 and evolution the central MHC. AB - The two classes of human complement component C4 proteins C4A and C4B manifest differential chemical reactivities and binding affinities towards target surfaces and complement receptor CR1. There are multiple, polymorphic allotypes of C4A and C4B proteins. A complex multiplication pattern of C4A and C4B genes with variations in gene size, gene dosage and flanking genes exists in the population. This is probably driven by the selection pressure to respond to a great variety of parasites efficiently and effectively, which the bony fish achieved through the multiplication and diversification of the related complement C3 proteins. Complement C4, C3 and C5 belong to the alpha2 macroglobulin protein family but acquired specific features that include an anaphylatoxin domain, a netrin (NTR) domain, and stretches of basic residues for proteolytic processings to form multiple chain structures. Complement C3 and C4 are important in the innate immune response as they opsonize parasites for phagocytosis. The emergence of complement C3 predates proteins involved in the adaptive immune response as C3 is present in deuterostome invertebrates such as echinoderms. The human C4 genes are located in the central MHC at chromosome 6p21.3. C3 and C5 are located at chromosome 19 and 9, respectively, with representatives of the other groups of genes paralogous to the MHC at 19p13.1-p13.3, 1q21-25, and 9q33-34. The central MHC also contains genes for complement components C2 and Bf. These genes appear to have similar evolutionary histories to C3/C4/C5 and are used here to illustrate stepwise processes resulting in co-location of diverse domains, chromosomal duplication, local segmental duplication and divergence of sequence and function. This model of evolution is useful in the investigation of innate and acquired immunity and in seeking explanations for diseases associated with MHC ancestral haplotypes. PMID- 11487476 TI - Genetic organization of the human MHC class III region. AB - The human major histocompatibility complex (MHC), or human leukocyte antigen (HLA) region, encompasses over 4 Mb of DNA on the short arm of chromosome 6 and is traditionally divided into the class I, class II and class III regions. The MHC has now been entirely sequenced and ~220 genes have been defined of which ~62 are in the class III region. It is becoming clear that many of the latter encode proteins that are likely to be involved in the immune and inflammatory responses. The MHC is known to contribute to a large number of immune-related disorders including insulin dependent diabetes mellitus, rheumatoid arthritis, common variable immunodeficiency and IgA deficiency and there is growing evidence that genes within the class III region are important in determining susceptibility to many of these complex conditions. Genes in the class III region have also been implicated in a number of non-immune-related diseases such as congenital adrenal hyperplasia and sialidosis. Now that the full gene content of the class III region is known the stage is set for the identification and characterisation of candidate disease genes, which will allow greater understanding of the causes of many MHC-linked diseases and thus aid the development of improved treatments. PMID- 11487477 TI - The bromodomain: a regulator of ATP-dependent chromatin remodeling? AB - In eukaryotes, processes requiring access to DNA are inhibited by the structural packaging of the genome. A number of specialized ATP-dependent chromatin remodeling enzymes have evolved to overcome this inhibition. One subset of these enzymes, SWI/SNF, plays a critical role in the regulation of transcription, often functioning in concert with nuclear histone acetyltransferases (HATs). It remains unknown how these activities are coordinated. However, recent results revealing that the bromodomain, a motif common in these remodeling factors, constitutes an acetyl-lysine binding domain might provide insight into this process. Bromodomains may serve a role analogous to the signal transduction SH2 domain, by providing a means to recruit remodeling complexes to acetylated chromatin regions or to allosterically modify their function post-recruitment. PMID- 11487478 TI - Developing a pediatric outpatient transplantation program. The Children's Memorial Hospital experience. AB - We describe the development of a pediatric outpatient transplant program and our initial experience with autologous and allogeneic transplants performed partially or completely in the outpatient setting. Forty-eight autologous and seven allogeneic transplants have been performed in our institution in the outpatient setting between June 1994 and July 2000. The ablation used for the autologous outpatient transplants was VP-16 1000 mg/m2/ day as a continuous infusion for 2 days and Carboplatinum 667mg/m2/day for 2 days. The autologous inpatient transplants received Thio-tepa 300-mg/ m2per day x 3 days and cyclophosphamide 60 mg/kg/day for 4 days. For those patients who received an immune-ablative allogeneic outpatient transplant, the regimen consisted of Fludarabine 30 mg/m2/day for 6 days, followed by busulfan for children less than five years of age 1 mg/kg/dose x 8 doses and for those five years and older 0.8 mg/kg/dose x 8 doses, followed by ATG 40mg/kg/day x 4 days. Engraftment was complete in all transplants achieving an ANC >500 for the outpatient transplant in 15 days (10 35) vs. the inpatient in 15 days (14-58). This was not statistically significant. They achieved un-sustained platelets >20.0 by day 19 (14-58) for the outpatients and day 32 10-64) for the inpatient. The allogeneic immune ablative transplants were considered engrafted when their VNTRs were greater that 30% which was achieved at a median of 13 days (10-27). The economic data showed a statistically significant decrease in charges and direct costs between the outpatient (median charges $30 775, direct costs $8 389) and the inpatient (median charges $99 838, direct costs $42 757) transplants (p0.001). There was no difference in morbidity and mortality between the two groups but the use of empiric amphotericin B was markedly decreased in the outpatient transplants. In conclusion it is feasible and less costly to perform autologous hematopoietic stem cell transplants in the outpatient setting with no increase in morbidity and mortality. For the allogeneic transplants there is not yet enough data to establish a similar conclusion. PMID- 11487479 TI - Endocrine complications of pediatric stem cell transplantation. AB - Abnormalities of endocrine function and growth are common following stem cell transplantation in the pediatric/adolescent population. Impaired linear growth and adult short stature are associated with younger age at transplant, use of TBI and prior cranial irradiation, and development of chronic GvHD. Primary hypothyroidism is the most common abnormality of the thyroid and is observed in 10-28% of cases following fractionated TBI. Autoimmune hyperthyroidism has also been described post-stem cell transplant and most often results from adoptive transfer of abnormal clones of T or B cells from donor to recipient. Gonadal dysfunction is extremely prevalent and includes oligo-azoospermia in the majority of males treated with TBI, and primary ovarian failure in most women treated with TBI or Busulfan/Cyclophosphamide. Leydig cell function, however, is retained in most males treated with standard forms of cytoreduction. Many patients demonstrate reduced bone mineral density and are at risk of developing osteoporosis in the future. PMID- 11487480 TI - CFTR and pseudomonas infections in cystic fibrosis. AB - Pseudomonas aeruginosa is a significant threat to human health as it is frequently recalcitrant to conventional antibacterial therapy. This ubiquitous gram-negative bacterium is notorious for its nutritional and ecological flexibility and its resistance to both antibiotic treatments and sanitary measures. These properties contribute to its prominence as a leading source of opportunistic nosocomial (hospital acquired) and a less appreciated, but significant cause of community acquired infections. P. aeruginosa remains a considerable problem for patients with burns, neutropenic individuals, and cystic fibrosis patients (CF). In this review, we will address the current issues in P. aeruginosa infections in CF. A major emphasis will be placed on the factors predisposing CF patients to colonization with P. aeruginosa. PMID- 11487481 TI - The association of MHC genes with autism. AB - Several immune abnormalities have been noted in autistic subjects. These associations have been extended to the Major Histocompatibility Complex (MHC), a section of DNA remarkable for the number of encoded proteins with immunological functions. The strongest MHC association identified thus far is for the null allele of C4B in the class III region. The complex allelic composition of C4 as determined by immunoelectrophoresis is discussed. Low levels of C4 resulting from the null allele may be important in disease pathogenesis especially since C4 has been identified in developing brain neurons. The DNA region just telomeric to C4 has several genes including tumor necrosis factor which encode proteins with immunological functions. These proteins may act in concert with C4 in disease contribution and the genes should be more closely examined. PMID- 11487482 TI - Patient survival after human albumin administration. A meta-analysis of randomized, controlled trials. AB - PURPOSE: To test the hypothesis that albumin administration is not associated with excess mortality. DATA SOURCES: Computer searches of the MEDLINE and EMBASE databases, the Cochrane Library, and Internet documents; hand searching of medical journals; inquiries to investigators and medical directors; and review of reference lists. STUDY SELECTION: Randomized, controlled trials comparing albumin therapy with crystalloid therapy, no albumin, or lower doses of albumin. DATA EXTRACTION: Two investigators independently extracted data. The primary end point was relative risk for death. Criteria used to assess methodologic quality were blinding, method of allocation concealment, presence of mortality as a study end point, and crossover. Small-trial bias was also investigated. DATA SYNTHESIS: Fifty-five trials involving surgery or trauma, burns, hypoalbuminemia, high-risk neonates, ascites, and other indications were included. Albumin administration did not significantly affect mortality in any category of indications. For all trials, the relative risk for death was 1.11 (95% CI, 0.95 to 1.28). Relative risk was lower among trials with blinding (0.73 [CI, 0.48 to 1.12]; n = 7), mortality as an end point (1.00 [CI, 0.84 to 1.18]; n = 17), no crossover (1.04 [CI, 0.89 to 1.22]; n = 35), and 100 or more patients (0.94 [CI, 0.77 to 1.14]; n = 10). In trials with two or more such attributes, relative risk was further reduced. CONCLUSIONS: Overall, no effect of albumin on mortality was detected; any such effect may therefore be small. This finding supports the safety of albumin. The influence of methodologic quality on relative risk for death suggests the need for further well-designed clinical trials. PMID- 11487483 TI - The effect of two different dosages of intravenous immunoglobulin on the incidence of recurrent infections in patients with primary hypogammaglobulinemia. A randomized, double-blind, multicenter crossover trial. AB - BACKGROUND: In patients with hypogammaglobulinemia, substitution with immunoglobulin is the treatment of choice to reduce both frequency and severity of bacterial infections. Even with treatment, however, infections still occur in these patients. OBJECTIVE: To determine whether doubling the standard dose of intravenous immunoglobulin would affect the incidence and duration of infections. DESIGN: Multicenter, double-blind, randomized, crossover study. SETTING: 15 outpatient clinics in the Netherlands. PATIENTS: 43 patients with primary hypogammaglobulinemia, 41 of whom completed the protocol. INTERVENTION: Patients received standard-dose immunoglobulin therapy for 9 months, followed by a 3-month washout period, and high-dose intravenous immunoglobulin therapy for 9 months, or vice versa. MEASUREMENTS: The primary outcome measures were total number and duration of infections. Other measures were periods of fever, hospital admissions, use of antibiotics, absence from school or work, and trough levels of serum immunoglobulin. Side effects from the study medication were also recorded. RESULTS: Compared with the standard dose of intravenous immunoglobulin (adults, 300 mg/kg of body weight every 4 weeks; children, 400 mg/kg every 4 weeks), high dose therapy (adults, 600 mg/kg every 4 weeks; children, 800 mg/kg every 4 weeks) significantly reduced the number (3.5 vs. 2.5 per patient; P = 0.004) and duration (median, 33 days vs. 21 days; P = 0.015) of infections. Trough levels of IgG increased significantly during high-dose therapy. The incidence and type of side effects did not differ significantly for the two dosages. CONCLUSION: In patients with hypogammaglobulinemia, doubling the standard dose of intravenous immunoglobulin significantly reduced the number and duration of infections. PMID- 11487484 TI - The effect of vancomycin and third-generation cephalosporins on prevalence of vancomycin-resistant enterococci in 126 U.S. adult intensive care units. AB - BACKGROUND: Patient-specific risk factors for acquisition of vancomycin-resistant enterococci (VRE) among hospitalized patients are becoming well defined. However, few studies have reported data on the institutional risk factors, including rates of antimicrobial use, that predict rates of VRE. Identifying modifiable institutional factors can advance quality-improvement efforts to minimize hospital-acquired infections with VRE. OBJECTIVE: To determine the independent importance of any association between antimicrobial use and risk factors for nosocomial infection on rates of VRE in intensive care units (ICUs). DESIGN: Prospective ecologic study. SETTING: 126 adult ICUs from 60 U.S. hospitals from January 1996 through July 1999. PATIENTS: All patients admitted to participating ICUs. MEASUREMENTS: Monthly use of antimicrobial agents (defined daily doses per 1000 patient-days), nosocomial infection rates, and susceptibilities of all tested enterococci isolated from clinical cultures. RESULTS: Prevalence of VRE (median, 10%; range, 0% to 59%) varied by type of ICU and by teaching status and size of the hospital. Prevalence of VRE was strongly associated with VRE prevalence among inpatient non-ICU areas and outpatient areas in the hospital, ventilator-days per 1000 patient-days, and rate of parenteral vancomycin use. In a weighted linear regression model controlling for type of ICU and rates of VRE among non-ICU inpatient areas, rates of vancomycin use (P < 0.001) and third generation cephalosporin use (P = 0.02) were independently associated with VRE prevalence. CONCLUSIONS: Higher rates of vancomycin or third-generation cephalosporin use were associated with increased prevalence of VRE, independent of other ICU characteristics and the endemic VRE prevalence elsewhere in the hospital. Decreasing the use rates of these antimicrobial agents could reduce rates of VRE in ICUs. PMID- 11487485 TI - A prospective study of Helicobacter pylori seropositivity and the risk for future myocardial infarction among socioeconomically similar U.S. men. AB - BACKGROUND: The role of Helicobacter pylori as a determinant of cardiovascular disease is controversial. OBJECTIVE: To determine whether previous exposure to H. pylori is associated with an increased risk for myocardial infarction. DESIGN: Prospective case-control study. SETTING: Physicians' Health Study. PARTICIPANTS: Initially healthy U.S. men. MEASUREMENTS: Titers of IgG antibody against H. pylori and several inflammatory markers were measured in baseline blood samples obtained from 445 men who subsequently had a myocardial infarction (case patients) and 445 men matched for age and smoking status who remained free of vascular disease (controls) during a mean follow-up of 8.9 years. RESULTS: Baseline seropositivity was similar among case-patients and controls (43.4% vs. 44.3%; rate ratio, 0.96 [95% CI, 0.7 to 1.3]). Minimal evidence of association was found between magnitude of seropositivity and subsequent risk and between seropositivity and levels of the inflammatory biomarkers. CONCLUSION: In a socioeconomically homogeneous population, we found limited evidence of association between H. pylori exposure and risk for future myocardial infarction. PMID- 11487486 TI - Varieties of healing. 1: medical pluralism in the United States. AB - Medicine has become interested in unconventional healing practices, ostensibly because of recent demographic research that reveals a thriving medical market of multiple options. This essay presents a historical overview of medical pluralism in the United States. Consistent evidence is examined suggesting that unconventional medicine has been a persistent presence in U.S. health care. Despite parallels with the past, the recent widespread interest in alternative medicine also represents a dramatic reconfiguration of medical pluralism-from historical antagonism to what might arguably be described as a topical acknowledgment of postmodern medical diversity. This recent shift may have less to do with acknowledging "new" survey data than with representing shifts in medicine's institutional authority in a consumer-driven health care environment. This essay is an introduction to a discussion of a taxonomy of contemporary U.S. medical pluralism, which also appears in this issue. PMID- 11487487 TI - Varieties of healing. 2: a taxonomy of unconventional healing practices. AB - The first of two essays in this issue demonstrated that the United States has had a rich history of medical pluralism. This essay seeks to present an overview of contemporary unconventional medical practices in the United States. No clear definition of "alternative medicine" is offered because it is a residual category composed of heterogeneous healing methods. A descriptive taxonomy of contemporary unconventional healing could be more helpful. Two broad categories of unconventional medicine are described here: a more prominent, "mainstream" complementary and alternative medicine (CAM) and a more culture-bound, "parochial" unconventional medicine. The CAM component can be divided into professional groups, layperson-initiated popular health reform movements, New Age healing, alternative psychological therapies, and non-normative scientific enterprises. The parochial category can be divided into ethno-medicine, religious healing, and folk medicine. A topologic examination of U.S. health care can provide an important conceptual framework through which health care providers can understand the current situation in U.S. medical pluralism. PMID- 11487488 TI - Colloid use for fluid resuscitation: evidence and spin. PMID- 11487489 TI - Complementary and alternative medicine--an Annals series. PMID- 11487490 TI - Physician-assisted suicide. AB - Medical professional codes have long prohibited physician involvement in assisting a patient's suicide. However, despite ethical and legal prohibitions, calls for the liberalization of this ban have grown in recent years. The medical profession should articulate its views on the arguments for and against changes in public policy and decide whether changes are prudent. In addressing such a contentious issue, physicians, policymakers, and society must fully consider the needs of patients, the vulnerability of particular patient groups, issues of trust and professionalism, and the complexities of end-of-life health care. Physician-assisted suicide is prominent among the issues that define our professional norms and codes of ethics. The American College of Physicians American Society of Internal Medicine (ACP-ASIM) does not support the legalization of physician-assisted suicide. The routine practice of physician assisted suicide raises serious ethical and other concerns. Legalization would undermine the patient-physician relationship and the trust necessary to sustain it; alter the medical profession's role in society; and endanger the value our society places on life, especially on the lives of disabled, incompetent, and vulnerable individuals. The ACP-ASIM remains thoroughly committed to improving care for patients at the end of life. PMID- 11487491 TI - Cost-effectiveness of screening for colorectal cancer. PMID- 11487492 TI - Cost-effectiveness of screening for colorectal cancer. PMID- 11487493 TI - Cost-effectiveness of screening for colorectal cancer. PMID- 11487495 TI - Physicians and patient spirituality. PMID- 11487497 TI - "Let me see if i have this right...": words that help build empathy. PMID- 11487500 TI - Glomerulonephritis associated with deficiencies and polymorphisms of complement components encoded in the class III region of the MHC. AB - An association between the complement system and immune complex glomerular disease in humans has long been recognized. In fact, much of our early understanding of the immunochemistry of complement activation developed with the study of acute and chronic glomerulonephritis (1). This manuscript will examine associations between glomerulonephritis and the three complement components encoded within the major histocompatibility complex: C4, C2, and factor B (B). The mechanisms by which deficiencies or polymorphisms in these components can mediate disease will be examined. PMID- 11487501 TI - Features of the two gene pairs RD-SKI2W and DOM3Z-RP1 located between complement component genes factor B and C4 at the MHC class III region. AB - Located at the 30 kb genomic region between complement factor B and component C4 are four ubiquitously expressed genes RD, SKI2W, DOM3Z and RP1. Besides RP1, the protein products of the other three genes each has highly conserved homologues or related proteins in lower eukaryotes, contains leucine zipper motifs for protein interaction, and plays important roles related to RNA metabolism. RD is a subunit of the negative transcription elongation factor, critical for the regulation of gene expression. It has an RNA recognition motif and 24 copies of Arg-Asp (RD) repeats. Ski2w is a nucleolar and cytoplasmic protein that has a putative RNA helicase domain. Fusion proteins of human Ski2w expressed in insect cells and bacteria have ATPase activity. The cytoplasmic protein of human Ski2w is associated with the polysomes and probably the 40S subunit of ribosomes. Ski2w is probably involved in the regulation of translation and RNA turnover. Dom3z is a nuclear protein whose yeast homologue forms a complex with an exoribonuclease. RP1 (or STK19) is a Ser/Thr nuclear protein kinase. No homologues of RP1 in lower eukaryotes have been discovered. Six polymorphic residues are present in human Ski2w and two in Dom3z. The potential roles of Ski2w and Dom3z on the clearance of degraded nuclear and cytoplasmic RNA raised their possibilities as susceptibility genes of systemic lupus erythematosus that is a disease with flawed processes in the removal of apoptotic materials. PMID- 11487502 TI - Seventh W.D.M. Paton Memorial Lecture. The man who never was--Walter Ernest Dixon FRS. PMID- 11487503 TI - Angiogenesis gene therapy to rescue ischaemic tissues: achievements and future directions. AB - Ischaemic diseases are characterized by an impaired supply of blood resulting from narrowed or blocked arteries that starve tissues of needed nutrients and oxygen. Coronary-atherosclerosis induced myocardial infarction is one of the leading causes of mortality in developed countries. Ischaemic disease also affects the lower extremities. Considerable advances in both surgical bypassing and percutaneous revascularization techniques have been reached. However, many patients cannot benefit from these therapies because of the extension of arterial occlusion and/or microcirculation impairment. Consequently, the need for alternative therapeutic strategies is compelling. An innovative approach consists of stimulating collateral vessel growth, a natural host defence response that intervenes upon occurrence of critical reduction in tissue perfusion (Isner & Asahara, 1999). This review will debate the relevance of therapeutic angiogenesis for promotion of tissue repair. The following issues will receive attention: (a) vascular growth patterns, (b) delivery systems for angiogenesis gene transfer, (c) achievements of therapeutic angiogenesis in myocardial and peripheral ischaemia, and (d) future directions to improve effectiveness and safety of vascular gene therapy. PMID- 11487504 TI - Voltage-dependent acceleration of Ca(v)1.2 channel current decay by (+)- and (-) isradipine. AB - Inhibition of Ca(v)1.2 by antagonist 1,4 dihydropyridines (DHPs) is associated with a drug-induced acceleration of the calcium (Ca(2+)) channel current decay. This feature is contradictorily interpreted as open channel block or as drug induced inactivation. To elucidate the underlying molecular mechanism we investigated the effects of (+)- and (-)-isradipine on Ca(v)1.2 inactivation gating at different membrane potentials. alpha(1)1.2 Constructs were expressed together with alpha(2)-delta- and beta(1a)- subunits in Xenopus oocytes and drug induced changes in barium current (I(Ba)) kinetics analysed with the two microelectrode voltage clamp technique. To study isradipine effects on I(Ba) decay without contamination by intrinsic inactivation we expressed a mutant (V1504A) lacking fast voltage-dependent inactivation. At a subthreshold potential of -30 mV a 200-times higher concentration of (-)-isradipine was required to induce a comparable amount of inactivation as by (+)-isradipine. At +20 mV the two enantiomers were equally efficient in accelerating the I(Ba) decay. Faster recovery from (-)- than from (+)-isradipine-induced inactivation at -80 mV in a Ca(v)1.2 construct (tau((-)-isr.(Cav1.2))=0.74 s50% of cultured neurons (EC(50)s 24 mu ml(-1) and 1.9 microM, respectively), assessed using Fura-2 AM. The PAR1 agonist completely desensitized responses to thrombin, indicating that thrombin stimulates neurons through PAR1. Injection of TF-NH(2) into the rat paw stimulated a marked and sustained oedema. An NK1R antagonist and ablation of sensory nerves with capsaicin inhibited oedema by 44% at 1 h and completely by 5 h. In wild-type but not PAR1(-/-) mice, TF-NH(2) stimulated Evans blue extravasation in the bladder, oesophagus, stomach, intestine and pancreas by 2 - 8 fold. Extravasation in the bladder, oesophagus and stomach was abolished by an NK1R antagonist. Thus, thrombin cleaves PAR1 on primary spinal afferent neurons to release SP, which activates the NK1R on endothelial cells to stimulate gap formation, extravasation of plasma proteins, and oedema. In intact tissues, neurogenic mechanisms are predominantly responsible for PAR1-induced oedema. PMID- 11487507 TI - 2,5-Di-t-butyl-1,4-benzohydroquinone (BHQ) inhibits vascular L-type Ca(2+) channel via superoxide anion generation. AB - The aim of the present study was to investigate the effects of 2,5-di-t-butyl-1,4 benzohydroquinone (BHQ), an inhibitor of the sarco-endoplasmic reticulum Ca(2+) ATPase (SERCA), on the whole-cell voltage-dependent L-type Ca(2+) current (I(Ca(L))) of freshly isolated smooth muscle cells from the rat tail artery using the patch-clamp technique. BHQ, added to the perfusion solution, reduced I(Ca(L)) in a concentration- (IC(50)=66.7 microM) and voltage-dependent manner. This inhibition was only partially reversible. BHQ shifted the voltage dependence of the steady-state inactivation curve to more negative potentials by 7 mV in the mid-potential of the curve, without affecting the activation curve as well as the time course of I(Ca(L)) inactivation. Preincubation of the cells either with 10 microM cyclopiazonic acid, a SERCA inhibitor, or with 3 mM diethyldithiocarbamate, an inhibitor of intracellular superoxide dismutase (SOD), did not modify BHQ inhibition of I(Ca(L)). On the contrary, this effect was no longer evident when SOD (250 u ml(-1)) was added to the perfusion medium. Either in the presence or in the absence of cells, BHQ gave rise to superoxide anion formation, which was markedly inhibited by the addition of SOD. These results indicate that, at micromolar concentrations, BHQ inhibits vascular I(Ca(L)) by giving rise to the formation of superoxide anion which in turn impairs the channel function. PMID- 11487508 TI - Treatment of hippocampal neurons with cyclosporin A results in calcium overload and apoptosis which are independent on NMDA receptor activation. AB - Calcineurin is a ubiquitous calcium/calmodulin dependent protein phosphatase that has been shown to regulate the activity of ion channels, glutamate release, and synaptic plasticity. In the present study we show that CsA, a specific inhibitor of calcineurin, affects the survival of cultures developed from hippocampal dentate gyrus. Mixed neuronal-glial cultures exposed to 8 - 40 microM CsA undergo cell death characterized by apoptotic changes in cellular and nuclear morphology. TUNEL-positive staining was observed only in neurons that developed pyknotic morphology after treatment with 8 microM CsA for 24 - 72 h. Immunocytochemical staining with an anti-GFAP monoclonal antibody revealed that astrocytes from mixed neuronal/glial cultures were unaffected by exposure to CsA at doses toxic for neurons and all TUNEL-positive cells were neurons. MK-801, a noncompetitive inhibitor of glutamate receptor, does not inhibit the appearance of TUNEL positive neurons and apoptotic changes in nuclear morphology. Preincubation of cells with 8 microM CsA increased basal intracellular calcium level in time dependent manner and decreased relative calcium response to glutamate. Application of 1 microM MK-801 had no effect on CsA-induced changes in Ca(2+) level. Our findings suggest that the neuronal death after CsA treatment is not a result of glutamate excitotoxicity and the increase in intracellular calcium concentration in neurons is not dependent on calcium influx via NMDA channel. PMID- 11487509 TI - Direct effect of an acyl-CoA:cholesterol acyltransferase inhibitor, F-1394, on atherosclerosis in apolipoprotein E and low density lipoprotein receptor double knockout mice. AB - The acyl-CoA:cholesterol acyltransferase (ACAT) enzyme is thought to be responsible for foam cell formation and the subsequent progression of atherosclerosis. The apolipoprotein E and low density lipoprotein receptor double knockout (apoE/LDLr-DKO) mouse is an animal model that develops severe hyperlipidaemia and atherosclerosis. Here we have examined the effect of oral administration of an ACAT inhibitor, F-1394, on atherosclerosis in apoE/LDLr-DKO mice fed a regular chow diet. In en face analysis, a dose of 10, 30, or 100 mg kg(-1) day(-1) F-1394 for 10 weeks reduced the extent of lesions visible in the aorta by 24, 28 and 38%, respectively, as detected by staining with oil red O, without affecting serum cholesterol level in these mice. At the highest dose 100 mg kg(-1) day(-1) of F-1394, the reduction was statistically significant. For quantitative analysis of the cellular and non-cellular components comprising the lesions at the aortic sinus, the effects of an oral dose of 100 mg kg(-1) day(-1) F-1394 for 15 weeks were studied. There was a significant reduction (31.9%) in the oil-red O-stained area in cross-sections of the aortic sinus. In addition, the neointimal area, as well as levels of ACAT-1 protein tended to be decreased (15.2 and 25.8%, respectively, not significant). However, the areas containing macrophages, smooth muscle cells, and collagen were not affected by F-1394. In vitro, F-1394 attenuated foam cell formation in mouse peritoneal macrophages. These results indicate that ACAT may be primarily responsible for lipid accumulation in atherosclerotic lesions, and that its inhibition diminishes the lipid deposition via a direct effect on macrophages in the arterial wall. PMID- 11487510 TI - Effects of sufentanil and NMDA antagonists on a C-fibre reflex in the rat. AB - The effects of intravenous sufentanil and pre-administration of N-methyl-D aspartate (NMDA) receptor antagonists were tested on a reflex triggered by C fibre activation. The reflex was elicited by electrical stimulation of the sural nerve and recorded from the ipsilateral biceps femoris muscle in halothane anaesthetized rats either (1) with an intact neuraxis or (2) in which the brain had previously been transected at the level of the obex. All four doses of sufentanil (0.33, 0.6, 1 and 2 microg kg(-1)) elicited a depression of the reflex in a dose-dependent manner. However, following the expected depression, all doses of sufentanil elicited both facilitation of the reflex and tonic inter-stimulus discharges. The C-fibre reflex was not modified following intravenous ketamine (1 mg kg(-1)) or (+)-HA966 (5 or 10 mg kg(-1)) but, when administered 5 min before sufentanil, these drugs enhanced both the extent and the duration of the depression and strongly reduced the facilitations. In the obex-transected rats, the depressive effect of 1 microg kg(-1) sufentanil increased, while the facilitation of the C-fibre reflex and the tonic inter-stimulus discharges disappeared. Pre-administration of 10 mg kg(-1) (+)-HA966 reinforced and prolonged the depressive effect of sufentanil. These results extend previous studies suggesting the involvement of NMDA receptors in the spinal transmission of nociceptive signals. They illustrate the potential of spinal NMDA receptor blockade to both enhance the analgesic, and prevent the pro-nociceptive, effects of sufentanil. PMID- 11487511 TI - Vasorelaxant effect of nitric oxide releasing steroidal and nonsteroidal anti inflammatory drugs. AB - The effect of several nitric oxide releasing-non-steroidal anti-inflammatory drugs (NO-NSAID) and nitroprednisolone on blood vessel relaxation in vitro and in vivo was studied. Nitroflurbiprofen (NOF; EC(50), 688.8+/-93.8 microM), nitroaspirin (NOA; EC(50), 57.9+/-6.5 microM), nitroparacetamol (NOPARA; EC(50), 71.5+/-14.6 microM) and nitroprednisolone (EC(50), 15.1+/-1.4 microM) caused concentration-related relaxation of noradrenaline (NA)-contracted rat aortic rings. All NO releasing compounds tested were approximately three orders of magnitude less potent than sodium nitroprusside (SNP, EC(50), 35.7+/-3.5 nM). The vasorelaxant effect of NOF and NOPARA in the rat aorta was potentiated by zaprinast (5 microM) and reduced by ODQ (5 microM). Flurbiprofen and paracetamol (100 microM) caused minimal (<10%) relaxation of the rat aorta and did not affect the response to SNP. The effect of NOF was unchanged in the presence of L-NAME (100 microM; EC(30), 181.8+/-35.1 microM cf. EC(30), 125.1+/-17.0 microM, P>0.05) but increased by removal of the endothelium (EC(30), 164.3+/-26.3 microM cf. EC(50), 688.8+/-93.8 microM, P<0.05). NOF (0.1 - 50 microM) produced a small but not concentration-related vasodilation of the NA-preconstricted (i.e. "high tone") perfused rat mesentery preparation (cf. SNP, EC(30), 4.4+/-0.7 microM). In contrast, NOF (1 - 100 microM) produced concentration-related vasodilation of the "high tone" perfused rat kidney with an EC(50) of 33.1+/-4.4 microM. Neither NOF (74 mg kg(-1), i.p.) nor NOA (91.9 mg kg(-1), i.p.) nor equimolar doses of flurbiprofen (50 mg kg(-1), i.p.) or aspirin (50 mg kg(-1), i.p.) affected mean arterial blood pressure (MAP) or heart rate (HR) of pentobarbitone-anaesthetized rats over a 1 h period. NO-NSAID relax blood vessels in vitro by an NO-dependent mechanism. The absolute vasorelaxant effect of NO releasing drug varies greatly with the choice of compound and between blood vessel preparations. PMID- 11487512 TI - The anti-migraine 5-HT(1B/1D) agonist rizatriptan inhibits neurogenic dural vasodilation in anaesthetized guinea-pigs. AB - These studies investigated the pharmacology of neurogenic dural vasodilation in anaesthetized guinea-pigs. Following introduction of a closed cranial window the meningeal (dural) blood vessels were visualized using intravital microscopy and the diameter constantly measured using a video dimension analyser. Dural blood vessels were constricted with endothelin-1 (3 microg kg(-1), i.v.) prior to dilation of the dural blood vessels with calcitonin gene-related peptide (CGRP; 1 microg kg(-1), i.v.) or local electrical stimulation (up to 300 microA) of the dura mater. In guinea-pigs pre-treated with the CGRP receptor antagonist CGRP((8 37)) (0.3 mg kg(-1), i.v.) the dilator response to electrical stimulation was inhibited by 85% indicating an important role of CGRP in neurogenic dural vasodilation in this species. Neurogenic dural vasodilation was also blocked by the 5-HT(1B/1D) agonist rizatriptan (100 microg kg(-1)) with estimated plasma levels commensurate with concentrations required for anti-migraine efficacy in patients. Rizatriptan did not reverse the dural dilation evoked by CGRP indicating an action on presynaptic receptors located on trigeminal sensory fibres innervating dural blood vessels. In addition, neurogenic dural vasodilation was also blocked by the selective 5-HT(1D) agonist PNU-142633 (100 microg kg(-1)) but not by the 5-HT(1F) agonist LY334370 (3 mg kg(-1)) suggesting that rizatriptan blocks neurogenic vasodilation via an action on 5-HT(1D) receptors located on perivascular trigeminal nerves to inhibit CGRP release. This mechanism may underlie one of the anti-migraine actions of the triptan class exemplified by rizatriptan and suggests that the guinea-pig is an appropriate species in which to investigate the pharmacology of neurogenic dural vasodilation. PMID- 11487513 TI - Pharmacological characterization of muscarinic receptors in mouse isolated urinary bladder smooth muscle. AB - The pharmacological characteristics of muscarinic receptors in the male mice urinary bladder smooth muscle were studied. (+)-Cis-dioxolane, oxotremorine-M, acetylcholine, carbachol and pilocarpine induced concentration-dependent contractions of the urinary bladder smooth muscle (pEC(50)=6.6+/-0.1, 6.9+/-0.1, 6.7+/-0.1, 5.8+/-0.1 and 5.8+/-0.1, E(Max)=3.2+/-0.8 g, 2.7+/-0.4 g, 1.0+/-0.1 g, 2.7+/-0.3 and 0.9+/-0.2 g, respectively, n=4). These contractions were competitively antagonized by a range of muscarinic receptor antagonists (pK(B) values): atropine (9.22+/-0.09), pirenzepine (6.85+/-0.08), 4-DAMP (8.42+/-0.14), methoctramine (5.96+/-0.05), p-F-HHSiD (7.48+/-0.09), tolterodine (8.89+/-0.13), AQ-RA 741 (7.04+/-0.12), s-secoverine (8.21+/-0.09), zamifenacin (8.30+/-0.17) and darifenacin (8.70+/-0.09). In this tissue, the pK(B) values correlated most favourably with pK(i) values for these compounds at human recombinant muscarinic M(3) receptors. A significant correlation was also noted at human recombinant muscarinic m5 receptors given the poor discriminative ability of ligands between M(3) and m5 receptors. In recontraction studies, in which the muscarinic M(3) receptor population was decreased, and conditions optimized to study M(2) receptor activation, methoctramine exhibited an affinity estimate consistent with muscarinic M(3) receptors (pK(B)=6.23+/-0.14; pA(2)=6.16+/-0.03). Overall, these data study suggest that muscarinic M(3) receptors are the predominant, if not the exclusive, subtype mediating contractile responses to muscarinic agonists in male mouse urinary bladder smooth muscle. PMID- 11487514 TI - Combination of miglitol, an anti-diabetic drug, and nicorandil markedly reduces myocardial infarct size through opening the mitochondrial K(ATP) channels in rabbits. AB - The anti-diabetic drug miglitol, an alpha-glucosidase inhibitor, which is currently used clinically, reduces myocardial infarct size by reducing the glycogenolytic rate through inhibition of the alpha-1,6-glucosidase of glycogen debranching enzyme in the heart. Nicorandil, a K(ATP) channel opener with a nitrate-like effect, which is also currently used clinically, also reduces the infarct size. Therefore, we hypothesized that combination of nicorandil and submaximal dose of miglitol could markedly reduce myocardial infarct size more than miglitol or nicorandil alone, and investigated the mechanism for the infarct size-reducing effect. Japanese white rabbits without collateral circulation were subjected to 30 min coronary occlusion followed by 48 h reperfusion. Pre ischaemic treatment with submaximal dose of miglitol (5 mg kg(-1), i.v.) and nicorandil alone (100 microg kg(-1) min(-1) 5 min) moderately reduced the infarct size as a percentage of area at risk (24+/-4 and 25+/-4%, respectively), and 10 mg kg(-1) of miglitol markedly reduced the infarct size (15+/-2%) compared with the controls (42+/-2%). Combination of 5 mg kg(-1) of miglitol and nicorandil (100 microg kg(-1) min(-1) 5 min), and 10 mg kg(-1) of miglitol and nicorandil (100 microg kg(-1) min(-1) 5 min) significantly reduced the infarct size (13+/-4 and 12+/-3%, respectively) more than miglitol or nicorandil alone. Pretreatment with 5HD completely abolished the infarct size-reducing effect of 10 mg kg(-1) of miglitol alone (36+/-7%) and that of combination of 5 mg kg(-1) of miglitol and nicorandil (46+/-2%). Combination of nicorandil and submaximal dose of miglitol markedly reduced the myocardial infarct size more than miglitol or nicorandil alone. This effect was suggested to be related to the opening of mitochondrial K(ATP) channels. PMID- 11487515 TI - Haemodynamic effects of the bacterial quorum sensing signal molecule, N-(3 oxododecanoyl)-L-homoserine lactone, in conscious, normal and endotoxaemic rats. AB - N-acylhomoserine lactones (AHLs) are small, diffusible signalling molecules, employed by Gram-negative bacteria to coordinate gene expression with cell population density. Recent in vitro findings indicate that AHLs may function as virulence determinants per se, through modification of cytokine production by eukaryotic cells, and by stimulating the relaxation of blood vessels. In the present study, we assessed the influence of AHLs on cardiovascular function in conscious rats, and draw attention to the ability of the N-(3-oxododecanoyl)-L homoserine lactone (3-oxo-C12-HSL), a signal molecule produced by P. aeruginosa, to cause marked bradycardia. This bradycardic effect was blocked by atropine and atenolol, and did not occur in vitro. Furthermore, modification of the acyl side chain length resulted in the loss of activity, whereas removal of the homoserine lactone ring, did not. The bradycardic effect of 3-oxo-C12-HSL was also observed in endotoxaemic animals, albeit attenuated. In normal rats, 3-oxo-C12-HSL caused initial mesenteric and hindquarters vasoconstriction, but only slight, and delayed signs of vasodilatation in the renal and mesenteric vascular beds. Furthermore, administration of 3-oxo-C12-HSL (pre-treatment or 2 h post treatment) together with LPS, did not modify the established regional haemodynamic effects of the LPS, 6 h after the onset of its infusion. Our observations do not provide any clear evidence for an ability of 3-oxo-C12-HSL to modify the haemodynamic responses to LPS infusion. However, they are not inconsistent with the hypothesis that some of the cardiovascular sequelae of bacterial infection may be modulated by an influence of bacterial quorum sensing signalling molecules on the host. PMID- 11487516 TI - Regional localization and developmental profile of acetylcholinesterase-evoked increases in [(3)H]-5-fluororwillardiine binding to AMPA receptors in rat brain. AB - In addition to its role in hydrolyzing the neurotransmitter acetylcholine, the synaptically enriched enzyme acetylcholinesterase (AChE) has been reported to play an important role in the development and remodelling of neural processes and synapses. We have shown previously that AChE causes an increase in binding of the specific AMPA receptor ligand (S)-[(3)H]-5-fluorowillardiine ([(3)H]-FW) to rat brain membranes. In this study we have used quantitative autoradiography to investigate the regional distribution and age-dependence of AChE-evoked increases in the binding of [(3)H]-FW in rat brain. Pretreatment of rat brain sections with AChE caused a marked enhancement of [(3)H]-FW binding to many, but not all, brain areas. The increased [(3)H]-FW binding was blocked by the specific AChE inhibitor BW 284c51. The maximal potentiation of [(3)H]-FW binding occurred at different developmental age-points in different regions with a profile consistent with the peak periods for synaptogenesis in any given region. In addition to its effects on brain sections, AChE also strongly potentiated [(3)H]-FW binding to detergent solubilized AMPA receptors suggesting a direct action on the receptors themselves rather than an indirect effect on the plasma membrane. These findings suggest that modulation of AMPA receptors could provide one molecular mechanism for the previously reported effects of AChE on synapse formation, synaptic plasticity and neurodegeneration. PMID- 11487517 TI - Functional characterization of coronary vascular adenosine receptors in the mouse. AB - Coronary responses to adenosine agonists were assessed in perfused mouse and rat hearts. The roles of nitric oxide (NO) and ATP-dependent K(+) channels (K(ATP)) were studied in the mouse. Resting coronary resistance was lower in mouse vs rat, as was minimal resistance (2.2+/-0.1 vs 3.8+/-0.2 mmHg ml(-1) min(-1) g(-1)). Peak hyperaemic flow after 20 - 60 s occlusion was greater in mouse. Adenosine agonists induced coronary dilation in mouse, with pEC(50)s of 9.4+/-0.1 for 2-[p (2-carboxyethyl)phenethylamino]-5'-N-ethyl carboxamidoadenosine (CGS21680, A(2A) selective agonist), 9.3+/-0.1 for 5'-N-ethylcarboxamidoadenosine (NECA, A(1)/A(2) agonist), 8.4+/-0.1 for 2-chloroadenosine (A(1)/A(2) agonist), 7.7+/-0.1 for N(6) (R)-(phenylisopropyl)adenosine (R-PIA, A(1)/A(2B) selective), and 6.8+/-0.2 for adenosine. The potency order (CGS21680=NECA>2-chloroadenosine>R-PIA>adenosine) supports A(2A) adenosine receptor-mediated dilation in mouse coronary vessels. 0.2 - 2 microM of the A(2B)-selective antagonist alloxazine failed to alter CGS21680 or 2-chloroadenosine responses. pEC(50)s in rat were 6.7+/-0.2 for CGS21680, 7.3+/-0.1 for NECA, 7.6+/-0.1 for 2-chloroadenosine, 7.2+/-0.1 for R PIA, and 6.2+/-0.1 for adenosine (2-chloroadenosine>NECA=R-PIA>CGS21680> adenosine), supporting an A(2B) adenosine receptor response. NO-synthase antagonism with 50 microM N(G)-nitro L-arginine (L-NOARG) increased resistance by approximately 25%, and inhibited responses to CGS21680 (pEC(50)=9.0+/-0.1), 2 chloroadenosine (pEC(50)=7.3+/-0.2) and endothelial-dependent ADP, but not sodium nitroprusside (SNP). K(ATP) channel blockade with 5 microM glibenclamide increased resistance by approximately 80% and inhibited responses to CGS21680 in control (pEC(50)=8.3+/-0.1) and L-NOARG-treated hearts (pEC(50)=7.3+/-0.1), and to 2-chloroadenosine in control (pEC(50)=6.7+/-0.1) and L-NOARG-treated hearts (pEC(50)=5.9+/-0.2). In summary, mouse coronary vessels are more sensitive to adenosine than rat vessels. A(2A) adenosine receptors mediate dilation in mouse coronary vessels vs A(2B) receptors in rat. Responses in the mouse involve a sensitive NO-dependent response and K(ATP)-dependent dilation. PMID- 11487518 TI - Vascular pharmacology of BIIE0246, the first selective non-peptide neuropeptide Y Y(2) receptor antagonist, in vivo. AB - BIIE0246, a recently introduced non-peptide neuropeptide Y (NPY) Y(2) receptor antagonist, was pharmacologically characterized in vivo, on vascular responses evoked in the anaesthetized pig. The NPY Y(2) receptor agonist N acetyl[Leu(28)Leu(31)]NPY(24-36) evoked dose-dependent vasoconstriction in spleen. These vascular responses were potently and dose-dependently antagonized by BIIE0246. Significant inhibition was seen already at 1 nmol kg(-1), whereas at 100 nmol kg(-1) of BIIE0246 these responses were completely abolished. The ID(50) value for this antagonism was 2.1 nmol kg(-1). Peptide YY (PYY) evoked dose dependent vasoconstriction in both kidney and spleen, vascular responses mediated by the NPY Y(1) receptor and both NPY Y(1) and Y(2) receptors, respectively. Only the splenic response was inhibited by BIIE0246, the effect of which reached significance at 1 nmol kg(-1). Already 30 min after the last dose of BIIE0246 there was a significant recovery of the PYY-evoked splenic vasoconstriction, and a further 60 min later, this response was no longer significantly inhibited compared to control. BIIE0246 (100 nmol kg(-1)) did not affect renal and splenic vasoconstrictor responses either to the NPY Y(1) receptor agonist [Leu(31)Pro(34)]NPY, the alpha(1)-adrenoceptor agonist phenylephrine, the P2X(1) purinoceptor agonist alpha,beta-methylene ATP or angiotensin II, demonstrating both selectivity and specificity for the NPY Y(2) receptor in vivo. It is concluded that BIIE0246 is a highly potent and selective NPY Y(2) receptor antagonist, albeit with rather short duration of action, in vivo. BIIE0246 thus represents the first interesting tool for studies on NPY Y(2) receptor-mediated transmission in vivo. PMID- 11487519 TI - A comparison of effects measured with isotonic and isometric recording: I. Concentration-effect curves for agonists. AB - Concentration-effect curves were obtained with carbachol tested on isolated preparations of guinea-pig ileum taken from adjacent sites in the same animal, one recorded isotonically, the other isometrically: similar experiments were made with histamine as agonist and with carbachol on rat uterus (in oestrus). The position and steepness of the curves was expressed as the values of [EC(50)] and the exponent, P: with carbachol or histamine on guinea-pig ileum the curves were significantly steeper with isotonic recording (P<0.02, sign test) and displaced towards lower concentrations (P<0.005) but there were significant correlations (P<0.05) between values obtained with tissues from the same animal. The curves for carbachol on the rat uterus were very steep: with isotonic recording the exponent (often eight or more) was consistently higher than with isometric (P<0.001): there was no significant displacement but there was a significant correlation (P<0.05) between values of [EC(50)] obtained with tissues from the same animal. Although the results obtained by the two methods are different, they are correlated. These effects are to be expected because with isotonic recording there can be no change in length until the tension exceeds the load and the tissue bulk sets an upper limit to shortening: the range within which an effect can be measured (the "operational window") is smaller. The observed effects on [EC(50)] and P have been reproduced with theoretical data. PMID- 11487520 TI - A comparison of effects measured with isotonic and isometric recording: II. Concentration-effect curves for physiological antagonists. AB - If one drug, B, antagonizes another, A, by producing the opposite physiological effect, the antagonist concentration-effect curves should be affected by the recording system, which limits the range of agonist responses. With pieces of isolated guinea-pig ileum taken from adjacent parts of the same animal, one recorded isotonically, the other isometrically with the same load, the isotonic IC(50) values for (-)isoprenaline opposing carbachol or histamine were lower than the isometric values (P<0.01) but there was a significant correlation between them (P<0.01): the isotonic curves were steeper (P<0.01) and there were wider shifts in IC(50) before increasing the agonist reduced the maximum relaxation. In similar experiments with pieces of rat uterus in oestrus from the same animal, the concentration-effect curves for carbachol opposed by increasing concentrations of (-)isoprenaline or (-)adrenaline had slightly lower EC(50) values with isometric recording but there was a significant correlation (P<0.01) with isotonic values. The antagonist effect (ratio of the EC(50) relative to that for the control) was higher with isotonic recording (P<0.01 for (-)isoprenaline, P<0.025 for (-)adrenaline) and all (27) curves were steeper than the corresponding isometric curve (P<0.001). The influence of the method of recording on the results is expected from the narrower operational window and smaller upper limit to relaxation with isotonic recording. A way of obtaining measurements of IC(50) against a standard agonist effect is suggested in an Appendix. PMID- 11487521 TI - Phloretin differentially inhibits volume-sensitive and cyclic AMP-activated, but not Ca-activated, Cl(-) channels. AB - Some phenol derivatives are known to block volume-sensitive Cl(-) channels. However, effects on the channel of the bisphenol phloretin, which is a known blocker of glucose uniport and anion antiport, have not been examined. In the present study, we investigated the effects of phloretin on volume-sensitive Cl(-) channels in comparison with cyclic AMP-activated CFTR Cl(-) channels and Ca(2+) activated Cl(-) channels using the whole-cell patch-clamp technique. Extracellular application of phloretin (over 10 microM) voltage-independently, and in a concentration-dependent manner (IC(50) approximately 30 microM), inhibited the Cl(-) current activated by a hypotonic challenge in human epithelial T84, Intestine 407 cells and mouse mammary C127/CFTR cells. In contrast, at 30 microM phloretin failed to inhibit cyclic AMP-activated Cl(-) currents in T84 and C127/CFTR cells. Higher concentrations (over 100 microM) of phloretin, however, partially inhibited the CFTR Cl(-) currents in a voltage dependent manner. At 30 and 300 microM, phloretin showed no inhibitory effect on Ca(2+)-dependent Cl(-) currents induced by ionomycin in T84 cells. It is concluded that phloretin preferentially blocks volume-sensitive Cl(-) channels at low concentrations (below 100 microM) and also inhibits cyclic AMP-activated Cl( ) channels at higher concentrations, whereas phloretin does not inhibit Ca(2+) activated Cl(-) channels in epithelial cells. PMID- 11487522 TI - Tachykinin NK(2) receptors and enhancement of cholinergic transmission in the inflamed rat colon: an in vivo motility study. AB - In the gastrointestinal tract, tachykinin NK(2) receptors are localized both on smooth muscle and nerve fibres. NK(2) receptor antagonists reduce exaggerated intestinal motility in various diarrhoea models but the site of action contributing to this effect is unknown. In this study we investigated the effects of atropine (1.4 micromol kg(-1), i.v.), hexamethonium (13.5 micromol kg(-1), i.v.), and nepadutant (0.1 micromol kg(-1), i.v.), a selective tachykinin NK(2) receptor antagonist, on distension (0.5 and 1 ml)-, or irritation (acetic acid, 0.5 ml of 7.5% v v(-1))-induced motility in the rat distal colon in vivo. The effects of atropine, hexamethonium or N(omega)-nitro-L-argininemethylester (L NAME, 1.85 micromol kg(-1), i.v.) on [betaAla(8)]NKA(4-10) (10 nmol kg(-1), i.v.) induced colonic contractions were also investigated. When the colonic balloon was filled with a subthreshold volume (0.5 ml), the intraluminal instillation of acetic acid triggered a high-amplitude phasic colonic motility which was partially reduced by nepadutant and suppressed by either hexamethonium or atropine. Filling of the balloon with 1 ml evoked reflex (hexamethonium sensitive), atropine-sensitive phasic colonic motility: nepadutant had no significant effect on the distension-evoked motility. Neither hexamethonium nor atropine significantly reduced [betaAla(8)]NKA(4-10)-induced colonic contractions, whereas nepadutant suppressed them. Following L-NAME pretreatment, [betaAla(8)]NKA(4-10)-induced colonic contractions were inhibited by both atropine and hexamethonium. In hexamethonium-pretreated animals, an atropine sensitive component of [betaAla(8)]NKA(4-10)-induced colonic contractions was also evident. These results indicate that the application of irritants onto the colonic mucosa induces the release of endogenous tachykinins which enhance excitatory cholinergic mechanisms through the stimulation of NK(2) receptors. PMID- 11487523 TI - Neuroprotective and neurorescuing effects of isoform-specific nitric oxide synthase inhibitors, nitric oxide scavenger, and antioxidant against beta-amyloid toxicity. AB - Beta amyloid (Abeta) is implicated in Alzheimer's disease (AD). Abeta(1 - 42) (5, 10, or 20 microM) was able to increase NO release and decrease cellular viability in primary rat cortical mixed cultures. L-NOARG and SMTC (both at 10 or 100 microM) - type I NOS inhibitors - reduced cellular NO release in the absence of Abeta(1 - 42). At 100 microM, both drugs decreased cell viability. L-NIL (10 or 100 microM), and 1400W (1 or 5 microM) - type II NOS inhibitors - reduced NO release and improved viability when either drug was administered up to 4 h post Abeta(1 - 42) (10 microM) treatment. L-NOARG and SMTC (both at 10 or 100 microM) were only able to decrease NO release. Carboxy-PTIO or Trolox (both at 10 or 100 microM) - a NO scavenger and an antioxidant, respectively - increased viability when administered up to 1 h post Abeta(1 - 42) treatment. Either L-NIL (50 microM) or 1400W (3 microM) and Trolox (50 microM) showed synergistic actions. Peroxynitrite (100 or 200 microM) reduced cell viability. Viabilities were improved by L-NIL (100 microM), 1400W (5 microM), carboxy-PTIO (10 or 100 microM), and Trolox (10 or 100 microM). Hence, the data show that Abeta(1 - 42) induced NO release in neurons and glial cells, and that Abeta neurotoxicity is, at least in part, mediated by NO. NO concentration modulating compounds and antioxidant may have therapeutic importance in neurological disorders where oxidative stress is likely involved such as in AD. PMID- 11487524 TI - Inhibition of DNA synthesis in cultured hepatocytes by endotoxin-conditioned medium of activated stellate cells is transforming growth factor-beta and nitric oxide-independent. AB - Activated hepatic stellate cells play a major role in the pathophysiology of chronic liver disease. They can influence the metabolism of hepatocytes by producing a variety of cytokines and growth factors. Upon stimulation with endotoxin, stellate cells also synthesize nitric oxide (NO), a potent mediator of growth of several cell types including hepatocytes. We investigated the effect of serum-free medium conditioned by activated stellate cells in the absence and presence of endotoxin on NO and DNA synthesis in hepatocytes. Stellate cells and hepatocytes were isolated by enzymatic digestion of the liver. Stellate cells were cultured for 10 days after which the majority exhibited alpha-smooth muscle actin (a marker for activated cells); hepatocytes were used after overnight culture. While the medium conditioned by stellate cells in the absence of endotoxin stimulated DNA synthesis in hepatocytes, medium conditioned in its presence inhibited this process in an endotoxin concentration-dependent manner (10 - 1000 ng ml(-1)). Endotoxin-conditioned stellate cell medium also stimulated NO synthesis in hepatocytes; the effect was consistent with increased protein and mRNA expression of inducible NO synthase (iNOS). However, inhibition of DNA synthesis in hepatocytes caused by endotoxin-conditioned stellate cell medium was unaffected by the NOS inhibitor, L-N(G)-monomethylarginine (L-NMMA), guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), and neutralizing antibodies for TGF-beta, IL-1beta, IL-6 and TNF-alpha. These results indicate that factors other than these cytokines produced by activated stellate cells upon stimulation with endotoxin or by hepatocytes challenged with endotoxin conditioned stellate cell medium inhibit DNA synthesis in hepatocytes. PMID- 11487525 TI - An alternative pathway for metabolism of leukotriene D(4): effects on contractions to cysteinyl-leukotrienes in the guinea-pig trachea. AB - Contractions of guinea-pig tracheal preparations to cysteinyl-leukotrienes (LTC(4), LTD(4) and LTE(4)) were characterized in organ baths, and cysteinyl leukotriene metabolism was studied using radiolabelled agonists and RP-HPLC separation. In the presence of S-hexyl GSH (100 microM) the metabolism of [(3)H] LTC(4) into [(3)H]-LTD(4) was inhibited and the LTC(4)-induced contractions were resistant to CysLT(1) receptor antagonism but inhibited by the dual CysLT(1)/CysLT(2) receptor antagonist BAY u9773 (0.3 - 3 microM) with a pA(2) value of 6.8+/-0.2. In the presence of L-cysteine (5 mM), the metabolism of [(3)H]-LTD(4) into [(3)H]-LTE(4) was inhibited and the LTD(4)-induced contractions were inhibited by the CysLT(1) receptor antagonist ICI 198,615 (1 - 10 nM) with a pA(2)-value of 9.3+/-0.2. However, at higher concentrations of ICI 198,615 (30 - 300 nM) a residual contraction to LTD(4) was unmasked, and this response was inhibited by BAY u9773 (1 - 3 microM). In the presence of the combination of S-hexyl GSH with L-cysteine, the LTD(4)-induced contractions displayed the characteristics of the LTC(4) contractile responses, i.e. resistant to CysLT(1) receptor antagonism, increased maximal contractions and slower time course. This qualitative change of the LTD(4)-induced contraction was also observed in the presence of S-decyl GSH (100 microM), GSH (10 mM) and GSSG (10 mM). S-hexyl GSH, S-decyl GSH, GSH and GSSG all stimulated a formation of [(3)H] LTC(4) from [(3)H]-LTD(4). In conclusion, GSH and GSH-related compounds changed the pharmacology of the LTD(4)-induced contractions by stimulating the conversion of LTD(4) into LTC(4). Moreover, the results indicate that, in addition to the metabolism of LTC(4) into LTD(4) and LTE(4), also the formation of LTC(4) from LTD(4) may regulate cysteinyl-leukotriene function. PMID- 11487526 TI - Further investigations into the endothelium-dependent hyperpolarizing effects of bradykinin and substance P in porcine coronary artery. AB - In porcine coronary arteries, smooth muscle hyperpolarizations produced by the nitric oxide donor, NOR-1, and the prostacyclin analogue, iloprost, were compared with those induced by substance P and bradykinin and attributed to the endothelium-derived hyperpolarizing factor (EDHF). In the presence of 300 microM L-nitroarginine and 10 microM indomethacin, iloprost-induced hyperpolarizations were partially inhibited by 10 microM glibenclamide whereas those to NOR-1, substance P and bradykinin were unaffected. Hyperpolarizations produced by maximally-effective concentrations of NOR-1 and NS1619 were identical (to -65 mV). They were significantly less than those generated by either substance P or bradykinin (to approximately -80 mV) and were abolished by iberiotoxin 100 nM, a concentration which had essentially no effect on responses to substance P or bradykinin. Incubation of segments of intact arteries for 16 - 22 h in bicarbonate-buffered Krebs solution had little effect on EDHF responses to substance P or bradykinin. In contrast, after incubation for this period of time in HEPES-buffered Tyrode solution or Krebs containing 10 mM HEPES the EDHF response to substance P was abolished and that to bradykinin was markedly reduced. The residual bradykinin-induced hyperpolarization following incubation in Tyrode solution was inhibited by iberiotoxin and by 10 microM 17-octadecynoic acid. We conclude that substance P activates only the EDHF pathway in the presence of nitric oxide synthase and cyclo-oxygenase inhibitors. Incubation in HEPES-buffered Tyrode solution abolishes the EDHF responses to substance P and bradykinin to reveal an additional hyperpolarizing mechanism, associated with the opening of K(+) channels, activated only by bradykinin. PMID- 11487527 TI - Absence of ligand-induced regulation of kinin receptor expression in the rabbit. AB - The induction of B(1) receptors (B(1)Rs) and desensitization or down-regulation of B(2) receptors (B(2)Rs) as a consequence of the production of endogenous kinins has been termed the autoregulation hypothesis. The latter was investigated using two models based on the rabbit: kinin stimulation of cultured vascular smooth muscle cells (SMCs) and in vivo contact system activation (dextran sulphate intravenous injection, 2 mg kg(-1), 5 h). Rabbit aortic SMCs express a baseline population of B(1)Rs that was up-regulated upon interleukin-1beta treatment ([(3)H]-Lys-des-Arg(9)-BK binding or mRNA concentration evaluated by RT - PCR; 4 or 3 h, respectively). Treatment with B(1)R or B(2)R agonists failed to alter B(1)R expression under the same conditions. Despite consuming endogenous kininogen (assessed using the kinetics of immunoreactive kinin formation in the plasma exposed to glass beads ex vivo) and producing hypotension mediated by B(2)Rs in anaesthetized rabbits, dextran sulphate treatment failed to induce B(1)Rs in conscious animals (RT - PCR in several organs, aortic contractility). By contrast, lipopolysaccharide (LPS, 50 microg kg(-1), 5 h) was an effective B(1)R inducer (kidney, duodenum, aorta) but did not reduce kininogen reserve. We tested the alternate hypothesis that endogenous kinin participate in LPS induction of B(1)Rs. Kinin receptor antagonists (icatibant combined to B-9858, 50 microg kg(-1) of each) failed to prevent or reduce the effect of LPS on B(1)R expression. Dextran sulphate or LPS treatments did not persistently down-regulate vascular B(2)Rs (jugular vein contractility assessed ex vivo). The kinin receptor autoregulation hypothesis is not applicable to primary cell cultures derived from a tissue known to express B(1)Rs in a regulated manner (aorta). The activation of the endogenous kallikrein-kinin system is ineffective to induce B(1)Rs in vivo in an experimental time frame sufficient for B(1)R induction by LPS. PMID- 11487528 TI - Induction of cyclo-oxygenase-2 in human endothelial cells by SIN-1 in the absence of prostaglandin production. AB - Nitric oxide (NO) regulates cyclo-oxygenase (COX) activity in various cell systems and reports conflict in regard to its stimulatory versus inhibitory role. Incubation of human umbilical vein endothelial cells (HUVEC) with SIN-1 (3 morpholinosydnonimine), a donor of NO, resulted in a rapid and dose-dependent increase in the expression of COX-2 as analysed by Western and Northern blotting. Incubation of HUVEC with SIN-1 and interleukine (IL)-1alpha resulted in increased induction of COX-2 compared with IL-1alpha alone and corresponded to an additive effect. The COX-2 induction was dependent on a de novo synthesis since cycloheximide, an inhibitor of protein synthesis, blocked the enzyme expression. The increase in COX-2 expression was not accompanied by a corresponding change in prostaglandin (PG) production. However, the COX activity was partially recovered when immunoprecipitated COX-2 was incubated with arachidonic acid and haematin. Peroxynitrite, a highly reactive nitrogen molecule derived from the interaction of NO and superoxide anion, significantly increased COX-2 expression. Under these conditions and within the limit of detection of the antibody, selective antibody for nitrotyrosine failed to detect nitrated COX-2 in immunoprecipitated COX-2 when cells where incubated with SIN-1 or SIN-1+IL-1alpha. Ro 31-8220, a specific inhibitor of protein kinase (PK) C, blocked the induction of COX-2. Also, SB203580, the selective inhibitor of p38 MAP kinase, strongly blocked the induction of COX-2 by SIN-1 in the presence or absence of IL-1alpha, whereas the MEK-1 inhibitor, PD 98059, affected it to a lesser extent. These data demonstrate that SIN-1 induces COX-2 in HUVEC in the absence of PG formation and suggest a complex regulation of COX-2 expression and PG formation by NO in endothelial cells. PMID- 11487529 TI - Rho-dependent inhibition of the induction of connective tissue growth factor (CTGF) by HMG CoA reductase inhibitors (statins). AB - It was supposed that inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) reductase (statins) might inhibit the expression of the fibrosis-related factor CTGF (connective tissue growth factor) by interfering with the isoprenylation of Rho proteins. The human renal fibroblast cell line TK173 was used as an in vitro model system to study the statin-mediated modulation of the structure of the actin cytoskeleton and of the expression of CTGF mRNA. Incubation of the cells with simvastatin or lovastatin time-dependently and reversibly changed cell morphology and the actin cytoskeleton with maximal effects observed after about 18 h. Within the same time period, statins reduced the basal expression of CTGF and interfered with CTGF induction by lysophosphatidic acid (LPA) or transforming growth factor beta. Simvastatin and lovastatin proved to be much more potent than pravastatin (IC(50) 1 - 3 microM compared to 500 microM). The inhibition of CTGF expression was prevented when the cells were incubated with mevalonate or geranylgeranylpyrophosphate (GGPP) but not by farnesylpyrophosphate (FPP). Specific inhibition of geranylgeranyltransferase-I by GTI-286 inhibited LPA-mediated CTGF expression whereas an inhibitor of farnesyltransferases FTI-276 was ineffective. Simvastatin reduced the binding of the small GTPase RhoA to cellular membranes. The effect was prevented by mevalonate and GGPP, but not FPP. These data are in agreement with the hypothesis that interference of statins with the expression of CTGF mRNA is primarily due to interference with the isoprenylation of RhoA, in line with previous studies, which have shown that RhoA is an essential mediator of CTGF induction. The direct interference of statins with the synthesis of CTGF, a protein functionally related to the development of fibrosis, may thus be a novel mechanism underlying the beneficial effects of statins observed in renal diseases. PMID- 11487532 TI - Gender equity in biomedical science: comments from a lone female associate editor. PMID- 11487531 TI - Antithrombotic efficacy of a novel factor Xa inhibitor, FXV673, in a canine model of coronary artery thrombolysis. AB - We compared the antithrombotic efficacy of a potent factor Xa inhibitor, FXV673, to heparin and RPR109891, a GPIIb/IIIa antagonist, when used as adjunctive therapy in a canine model of rt-PA-induced coronary thrombolysis. Thrombus formation was induced by electrolytic injury to stenosed coronary artery. After thrombotic occlusion, a 135 min infusion of saline (n=8), FXV673 (10, 30 or 100 microg kg(-1)+1, 3, or 10 microg kg(-1) min(-1), respectively; n=8 per dose), heparin (60 u kg(-1)+0.7 u kg(-1) min(-1), n=8), or RPR109891 (30 microg kg( 1)+0.45 microg kg(-1) min(-1), n=8), was initiated. Aspirin (5 mg kg(-1), i.v.) was administered to all animals. Fifteen minutes after the start of drug infusion, rt-PA was administered (100 microg kg(-1)+20 microg kg(-1) min(-1) for 60 min). The incidence of reperfusion in the high dose FXV673 (8/8, 100%) was significantly greater than that in the heparin group (4/8, 50%), with a trend to faster reperfusion (23+/-5 min for FXV673 versus 41+/-11 min for heparin). Only 2/8 (25%) of the vessels reoccluded in the high dose FXV673 group, compared to 4/4 (100%) and 5/5 (100%) vessels in the heparin and RPR109891 groups, respectively (P<0.05). Throughout the protocol, blood flow was higher in the FXV673 treated group compared to other groups. FXV673 enhanced vessel patency in a dose-dependent manner. Compared to vehicle and heparin groups, the thrombus mass was decreased by 60% in the high dose FXV673. FXV673, heparin and RPR109891 increased the bleeding time by 2.7, 1.7 and 4 fold, and APTT by 2.8, 2.7 and 1.2 fold, respectively. In conclusion, FXV673 is more effective than heparin and at least as effective as RPR109891 when used as an adjunct during rt-PA-induced coronary thrombolysis. PMID- 11487530 TI - A comparison of allergen and polycation induced cutaneous responses in the rabbit. AB - Allergic inflammatory responses contribute to the symptoms of a number of diseases including atopic dermatitis, asthma and rhinitis. Cationic proteins are released from inflammatory cells and levels are known to be raised in disease states. Using an in vivo model of acute inflammation, we investigated the characteristics of cutaneous responses to antigen (Alternaria tenuis, AT) and poly-L-lysine (PLL, used as a paradigm for cationic proteins). We aimed to compare the inflammatory profile of cationic polypeptides and the allergic response and to identify similarities and differences between these responses. Responses to intradermal injection of the polycation, PLL and antigen were compared using radiolabelled protein ((125)I-bovine serum albumin, BSA) and cells ((111)In-neutrophils, PMN) to study plasma exudation (PE) and PMN accumulation (PMNA) in the skin of AT sensitized rabbits. Both PLL and antigen caused dose related increases in PE and PMNA. PE (and PMNA) responses to PLL were prolonged (up to 3 h), as were those to antigen. This is in contrast to PE responses to fMLP which were maximal at 45 min. In immunized animals, treated with colchicine (1 mg kg(-1) i.v.), PE responses to the directly acting mediator, bradykinin (BK), were not affected, whereas PE responses to the neutrophil dependent mediator, f-met-leu-phe (fMLP), were significantly (P<0.01) reduced. Antigen induced PE responses were significantly (50, 500 (P<0.05); 200 (P<0.01) p.n.u. site(-1)) inhibited by colchicine, but PLL-induced responses were not significantly affected. We conclude that although PLL-induced responses had a similar time course to those of antigen, some differences were observed between responses, which indicate that although polycations may contribute to allergic responses, these two responses are produced by distinct mechanisms. PMID- 11487533 TI - CCR9-positive lymphocytes and thymus-expressed chemokine distinguish small bowel from colonic Crohn's disease. AB - BACKGROUND & AIMS: Thymus-expressed chemokine (TECK) or CCL25) is selectively expressed in the small bowel (SB), where lamina propria lymphocytes (LPL) and intraepithelial leukocyte expressing the cognate chemokine receptor CCR9 predominate. We characterize the role of TECK and CCR9-expresing lymphocytes in small intestinal Crohn's disease. METHODS: CCR9 expression on lymphocytes from lamina propria, mesenteric lymph node, and peripheral blood was analyzed by flow cytometry and by Northern blotting for LPL. TECK expression was analyzed in inflamed SB and colon by reverse-transcription polymerase chain reaction and immunohistochemistry. RESULTS: The fraction of CCR9(+) T cells in inflamed SB was significantly lower than in uninvolved SB mucosa. In contrast, in peripheral blood lymphocytes, CCR9(+) lymphocytes were markedly elevated in patients with small bowel Crohn's or celiac disease, but not in patients with purely colonic Crohn's. Also, TECK expression is altered in inflamed small bowel, being intensely expressed in a patchy distribution in crypt epithelial cells in proximity to lymphocytic infiltrates. TECK is not expressed in either normal or inflamed colon. CONCLUSIONS: In SB immune-mediated diseases, there is repartitioning of CCR9(+) lymphocytes between SB and blood and an altered pattern of TECK expression in SB Crohn's. The TECK/CCR9 ligand/receptor pair may play an important role in the pathogenesis of SB Crohn's disease. PMID- 11487534 TI - The natural history of corticosteroid therapy for inflammatory bowel disease: a population-based study. AB - BACKGROUND & AIMS: The aim of this study was to determine the 1-year outcome after the first course of corticosteroids in an inception cohort of patients with inflammatory bowel disease. METHODS: All patients in Olmsted County, Minnesota, diagnosed with Crohn's disease (n = 173) or ulcerative colitis (n = 185) from 1970 to 1993 who were treated with systemic corticosteroids were identified (4 denied research authorization). Immediate outcome (30 days) and 1-year outcome after the first course of corticosteroids were determined. RESULTS: Seventy-four (43%) patients with Crohn's disease and 63 (34%) with ulcerative colitis were treated with corticosteroids. Immediate outcomes for Crohn's disease were complete remission in 43 (58%), partial remission in 19 (26%), and no response in 12 (16%). Immediate outcomes for ulcerative colitis were complete remission in 34 (54%), partial remission in 19 (30%), and no response in 10 (16%). One-year outcomes for Crohn's disease were prolonged response in 24 (32%), corticosteroid dependence in 21 (28%), operation in 28 (38%), and lost to follow-up in 1 (1%). One-year outcomes for ulcerative colitis were prolonged response in 31 (49%), corticosteroid dependence in 14 (22%), and operation in 18 (29%). CONCLUSIONS: Most patients with Crohn's disease and ulcerative colitis initially respond to corticosteroids. At 1 year, 32% of patients with Crohn's disease and 48% with ulcerative colitis are corticosteroid free without operation. PMID- 11487535 TI - Endoscopic and histologic evaluation together with symptom assessment are required to diagnose pouchitis. AB - BACKGROUND & AIMS: Pouchitis often is diagnosed based on symptoms alone. In this study, we evaluate whether symptoms correlate with endoscopic and histologic findings in patients with ulcerative colitis and an ileal pouch-anal anastomosis. METHODS: Symptoms, endoscopy, and histology were assessed in 46 patients using Pouchitis Disease Activity Index (PDAI). Patients were classified as either having pouchitis (PDAI score > or =7; N = 22) or as not having pouchitis (PDAI score <7; N = 24). RESULTS: Patients with pouchitis had significantly higher mean total PDAI scores, symptom scores, endoscopy scores, and histology scores. There was a similar magnitude of contribution of each component score to the total PDAI for the pouchitis group. Of note, 25% of patients with symptoms suggestive of pouchitis did not meet the PDAI diagnostic criteria for pouchitis. In both groups, the correlation coefficients between symptom, endoscopy, and histology scores were near zero (range, -0.26 to 0.20; P > 0.05). CONCLUSIONS: The symptom, endoscopy, and histology scores each contribute to the PDAI and appear to be independent of each other. Symptoms alone do not reliably diagnose pouchitis. PMID- 11487536 TI - A randomized placebo-controlled trial of a humanized monoclonal antibody to alpha4 integrin in active Crohn's disease. AB - BACKGROUND & AIMS: alpha4 integrins are important mediators of leukocyte migration across vascular endothelium. This pilot placebo-controlled study aimed to assess the safety and efficacy of natalizumab, a recombinant humanized monoclonal antibody to alpha4 integrin, in patients with mild to moderately active Crohn's disease. METHODS: Thirty patients with active Crohn's disease (Crohn's Disease Activity Index [CDAI] > or =151 and < or =450) received a 3 mg/kg infusion of natalizumab (n = 18) or placebo (n = 12) by double-blind randomization. The study's primary endpoint was change in CDAI at week 2. RESULTS: At week 2, the CDAI decreased significantly from baseline after infusion of natalizumab (mean 45 points) but not placebo (mean 11 points). Seven (39%) natalizumab-treated patients achieved remission at week 2, compared with 1 (8%) treated with placebo. In contrast, 4 (33%) of the placebo-treated patients required rescue medication by week 2, compared with 2 (11%) natalizumab-treated patients. Significant increases in circulating B and T lymphocytes were detected only after natalizumab administration. The frequency of commonly reported adverse events did not differ significantly between groups. CONCLUSIONS: A single 3-mg/kg natalizumab infusion was well tolerated by Crohn's disease patients, although the dose used may have been suboptimal. Elevated circulating lymphocyte levels after natalizumab suggest interrupted lymphocyte trafficking. Natalizumab therapy in active Crohn's disease merits further investigation. PMID- 11487537 TI - Risk of dysplasia in long-term ileal pouches and pouches with chronic pouchitis. AB - BACKGROUND & AIMS: Recent reports have suggested the mucosa of an ileal reservoir could be at risk of neoplasia. Risk factors may include the age of the pouch, chronic pouchitis, and previous colonic neoplasia. This study examined a group of such patients to determine the risk of dysplasia. METHODS: From a cohort of 1221 patients with ileal pouches, 171 patients with possible risk factors were selected. Successful contact was made with 138 patients who were invited for endoscopy and multiple biopsies. Biopsy specimens were stained with H&E and p53, scored for inflammatory changes including villous atrophy, and analyzed by flow cytometry. RESULTS: One hundred six patients took part and fell into 1 or more of the following clinical categories: chronic pouchitis (n = 34), pelvic pouch for > or =12 years (n = 42); Kock pouch for > or =14 years (n = 29), and neoplasia in colectomy specimen (n = 11). Thirty-three patients had severe villous atrophy. One patient of 106 (95% confidence interval, 0.9% +/- 1.6%) with a long-standing pouch had low-grade dysplasia that was multifocal. DNA analysis by flow cytometry showed aneuploidy in this patient and 2 others. CONCLUSIONS: These data suggest that the development of dysplasia in ileal pouches performed for ulcerative colitis is probably a rare event within 15-20 years of pouch surgery. PMID- 11487538 TI - Polymorphisms and colorectal tumor risk. AB - BACKGROUND & AIMS: Increasingly, studies of the relationship between common genetic variants and colorectal tumor risk are being proposed. To assess the evidence that any of these confers a risk, a systematic review and meta-analysis of published studies was undertaken. METHODS: Fifty studies of the effect of common alleles of 13 genes on risk were identified. To clarify the impact of individual polymorphisms on risk, pooled analyses were performed. RESULTS: Of the 50 studies identified, significant associations were seen in 16, but only 3 were reported in more than one study. Pooling studies, significant associations were only seen for 3 of the polymorphisms: adenomatosis polyposis coli (APC)-I1307K (odds ratio [OR] = 1.58, 95% confidence interval [CI]: 1.21-2.07); Harvey ras-1 variable number tandem repeat polymorphism (HRAS1-VNTR; OR = 2.50, 95% CI: 1.54 4.05); and methylenetetrahydrofolate reductase (MTHFR)(Val/Val) (OR = 0.76, 95% CI: 0.62-0.92). For tumor protein 53 (TP53), N-acetyl transferase 1 (NAT1), NAT2, glutathione-S transferase Mu (GSTM1), glutathione-S transferase Theta (GSTT1), and glutathione-S transferase Pi (GSTP1) polymorphisms, the best estimates are sufficient to exclude a 1.7-fold increase in risk of colorectal cancer. CONCLUSIONS: APC-I1307K, HRAS1-VNTR, and MTHFR variants represent the strongest candidates for low penetrance susceptibility alleles identified to date. Although their genotypic risks are modest, their high frequency in the population implies that they may well have considerable impact on colorectal cancer incidence. Determining precise risk estimates associated with other variants and gene-gene and gene-environment interactions will be contingent on further studies with sample sizes larger than typically used to date. PMID- 11487539 TI - Ki-ras proto-oncogene mutations in sporadic colorectal adenomas: relationship to histologic and clinical characteristics. AB - BACKGROUND & AIMS: [corrected] The goal of this study was to examine the relationship between Ki-ras mutations in colorectal adenomas and characteristics of both the subject (age, gender, and family/personal history of colonic neoplasia) and the adenoma (multiplicity, size, location, and histologic features). METHODS: Ki-ras mutations were detected by direct sequencing in 738 adenomatous polyps removed at baseline from 639 participants in a nutritional trial of adenoma recurrence. RESULTS: Ki-ras mutations were detected in 17.2% of the adenomas. Ki-ras mutations were unrelated to gender, family, or personal history of colonic neoplasia, location within the colorectum, or adenoma multiplicity, but were more common in older subjects (P = 0.01 for trend), in larger adenomas (P < 0.0001 for trend), in adenomas with villous histology (odds ratio [OR], 3.2; 95% confidence interval [CI], 2.1-4.9 vs. tubular), and in adenomas with high-grade dysplasia (32.0% vs. 13.6%; OR, 3.0; 95% CI, 1.9-4.6 vs. low-grade dysplasia). Multivariate analysis showed Ki-ras mutations to be independently associated with subject age (P = 0.01 for trend), tubulovillous/villous histology (OR, 2.3; 95% CI, 1.5-3.7), and high-grade dysplasia (OR, 1.9; 95% CI, 1.2-3.1). Adenoma size was not independently related to Ki-ras mutation. CONCLUSIONS: Ki-ras mutations are associated with the histologic features of adenoma progression (villous histology and high-grade dysplasia) rather than with adenoma growth. PMID- 11487540 TI - Helicobacter pylori infection in Swedish school children: lack of evidence of child-to-child transmission outside the family. AB - BACKGROUND & AIMS: Helicobacter pylori infection is mainly acquired in early childhood, but the exact routes of transmission remain elusive. To distinguish between risks of intrafamilial and extraneous child-to-child transmission, we studied H. pylori seroprevalence among Swedish school children with varying family backgrounds. METHODS: In a cross-sectional study, 695 of 858 (81%) 10-12 year-olds in 36 school classes in Stockholm donated blood and answered a questionnaire. Infection was detected by enzyme-linked immunosorbent assay and confirmed by immunoblot and urea breath test. RESULTS: Overall, 112 (16%) children were infected. The seroprevalence was 2% among 435 children with Scandinavian parents and 55% among 144 children with origin in high prevalence areas (Middle East and Africa). Among children born in Scandinavia, the odds ratios (adjusted for gender, socioeconomic status, and family size) for being seropositive were 39.1 (95% confidence interval, 16.7-91.3) and 5.6 (1.8-17.3) when having parents born in high and medium prevalence areas, respectively, relative to children with Scandinavian parents. Importantly, the prevalence of infection among the classmates was not a risk factor for H. pylori infection. CONCLUSION: Our data indicate that intrafamilial transmission is far more important than child-to-child transmission outside the family. The H. pylori prevalence in the parental generation may be a crucial determinant for the child's risk of contracting the infection. PMID- 11487541 TI - Sonic hedgehog regulates gastric gland morphogenesis in man and mouse. AB - BACKGROUND & AIMS: Gastric epithelial renewal is an asymmetric process. A stem cell located halfway up the tubular unit gives rise to both a basal gland region and a luminal pit compartment, but the mechanisms responsible for the maintenance of this asymmetry are obscure. We investigated whether Sonic hedgehog (Shh), an established polarizing signal protein during development, is expressed and functional in the adult human and murine stomach. METHODS: Expression of Shh and putative transcriptional targets was investigated using immunoblot and immunohistochemistry. Mice were treated with the Shh inhibitor cyclopamine and examined for expression levels of Shh targets and proliferation of gastric epithelial cells. RESULTS: Shh was expressed in the stomach. In cyclopamine treated mice, we observed decreased expression of HNF3beta, Islet (Isl)-1 and BMP4, 3 putative Shh target genes. Inhibition of Shh markedly enhanced gastric epithelial proliferation and affected the cell cycle of gastric epithelial gland cells, whereas pit cells remained unaffected. CONCLUSIONS: Shh controls the expression of at least 3 factors important for epithelial differentiation and is a negative regulator of gastric gland cell proliferation. Shh is a candidate polarizing signal in the maintenance of gastric pit-gland asymmetry. PMID- 11487542 TI - A critical role for IL-7R signaling in the development of Helicobacter felis induced gastritis in mice. AB - BACKGROUND & AIMS: Interleukin (IL)-7 is a critical cytokine in the development of T and B cells and is involved in gastrointestinal pathophysiology. The aim of this study was to investigate the involvement of IL-7 receptor (IL-7R) signaling in Helicobacter-induced gastritis. METHODS: C57BL/6 mice (n = 40) were inoculated with H. felis. Twenty mice were injected intraperitoneally with neutralizing IL 7R antibody (A7R34) every seventh day for 3 months. Histology, serum anti-H. felis antibody, and gene expression of IL-7, IL-7R, and proinflammatory cytokines in the gastric mucosa were evaluated. RESULTS: Seventeen of 20 (85%) infected mice without A7R34 developed severe atrophic gastritis, whereas there was no gastritis in A7R34-treated infected mice. There was no difference in the serum levels of anti-H. felis antibody between the 2 groups. IL-7, IL-7R, IL-1alpha, tumor necrosis factor alpha, and interferon gamma messenger RNA expressions were up-regulated in control infected mice, whereas only IL-7 messenger RNA was up regulated in A7R34-treated infected mice. Immunohistochemistry indicated positive cytoplasmic staining of IL-7 in the gastric epithelial cells. CONCLUSIONS: These data suggest a critical role for IL-7 receptor signaling in the development of Helicobacter-induced gastritis in mice. PMID- 11487543 TI - Intestinal epithelial cells secrete exosome-like vesicles. AB - BACKGROUND & AIMS: Given the observations that intestinal epithelial cells (IECs) can present antigens to CD4(+) T lymphocytes and that professional antigen presenting cells secrete exosomes (antigen-presenting vesicles), we hypothesized that IECs may secrete exosomes carrying molecules implicated in antigen presentation, which may be able to cross the basement membrane and convey immune information to noncontiguous immune cells. METHODS: Human IEC lines HT29-19A and T84-DRB1*0401/CIITA were grown on microporous filters. Release of exosomes under basal or inflammatory conditions was evaluated in conditioned apical and basolateral media after differential ultracentrifugations. Morphologic and biochemical characterization of exosomes was performed using immunoelectron microscopy, Western blotting, and matrix-assisted laser desorption ionization time of flight mass spectrometry. RESULTS: The intestinal cell lines released 30 90-nm-diameter vesicles from the apical and basolateral sides, and this release was significantly increased in the presence of interferon gamma. MHC class I, MHC class II, CD63, CD26/dipeptidyl-peptidase IV, and A33 antigen were present in epithelial-derived exosomes. CONCLUSIONS; Human IEC lines secrete exosomes bearing accessory molecules that may be involved in antigen presentation. These data are consistent with a model in which IECs may influence antigen presentation in the mucosal or systemic immune system independent of direct cellular contact with effector cells. PMID- 11487544 TI - Megacystis-microcolon-intestinal hypoperistalsis syndrome and the absence of the alpha3 nicotinic acetylcholine receptor subunit. AB - BACKGROUND & AIMS: The megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS) is a rare disease of childhood that presents early with intestinal hypoperistalsis, hydronephrosis, and hydroureters. Transgenic mice that lack the alpha3 subunit containing nicotinic acetylcholine (nAChR) have a phenotype similar to that of MMIHS. METHODS: We examined the expression of this subunit in control and MMIHS tissue derived from patients using in situ hybridization (ISH) and immunocytochemistry (ICC). RESULTS: In controls, both techniques showed a wide distribution of alpha3 nAChRs present in ganglion cells, muscle, and epithelium. By contrast, most MMIHS tissue gave negative staining with ISH and variable results with ICC. CONCLUSIONS: These observations are consistent with a lack of alpha3 nAChRs contributing to the pathogenesis of MMIHS. PMID- 11487545 TI - Somatostatin sst(2) receptor-mediated inhibition of mesenteric afferent nerves of the jejunum in the anesthetized rat. AB - BACKGROUND & AIMS: Octreotide inhibits visceral sensations in clinical studies, but the site of action and the receptor type(s) involved are unknown. Our aim was to investigate the effects of octreotide, the selective sst(2) receptor agonist (BIM 23027), and the sst(2) antagonist (Cyanamid154806) on the activity of mesenteric afferent fibers innervating the rat jejunum. Their effects were investigated on baseline discharge, mechanosensitivity, and responses to algesic chemicals. METHODS: Extracellular multiunit recordings of jejunal afferent nerve firing were made in pentobarbitone-anesthetized (60 mg/kg intraperitoneally) male Wistar rats. RESULTS: Octreotide and BIM23027 (0.001-100 microg/kg intravenously) each evoked a long-lasting inhibition of baseline discharge, which was blocked by cyanamid 154806 (3 mg/kg) and absent in chronically vagotomized animals. Afferent responses to bradykinin were also inhibited by an sst(2) receptor-mediated mechanism but were unaffected by vagotomy. Ramp distentions of the jejunum evoked a biphasic activation of afferent nerve discharge, the low threshold component of which was attenuated in vagotomized animals. Sst(2) receptor agonists significantly inhibited the mechanosensitivity of spinal, but not vagal, afferents. CONCLUSIONS: These data suggest that activation of somatostatin sst(2) receptors inhibit populations of mesenteric afferents likely to be involved in nociceptive transmission. PMID- 11487546 TI - Enhanced transepithelial antigen transport in intestine of allergic mice is mediated by IgE/CD23 and regulated by interleukin-4. AB - BACKGROUND & AIMS: We previously described a system for enhanced transepithelial transport of antigen in which both the amount of specific antigen and its rate of transport were dramatically increased in intestine of sensitized rats compared with controls. This study investigated the essential components mediating antigen uptake in mice genetically deficient for interleukin (IL)-4 or CD23. METHODS: Mice were actively or passively sensitized to horseradish peroxidase (HRP). Jejunal segments from control or sensitized mice were mounted in Ussing chambers and challenged with HRP from the luminal side. Tissues were processed for electron microscopy, and photomicrographs were analyzed for antigen uptake (location and area of HRP-containing endosomes). Immunohistochemistry and reverse transcription polymerase chain reaction were used to detect epithelial CD23 expression. RESULTS: Actively sensitized IL-4(+/+), but not IL-4(-/-) mice, displayed increased transepithelial antigen transport and CD23 expression on enterocytes. Passively sensitized IL-4(+/+) and IL-4(-/-) mice displayed elevated antigen transport after transfer of immune serum but not if the serum was depleted of immunoglobulin (Ig) E or IL-4. IL-4 added to cultured IEC-4 cells up regulated expression of CD23 messenger RNA. The augmented antigen uptake was inhibited by anti-CD23 and was absent in sensitized CD23(-/-) mice. CONCLUSIONS: Our studies indicate that IL-4 regulates IgE/CD23-mediated enhanced transepithelial antigen transport in sensitized mouse intestine. PMID- 11487547 TI - Randomized comparison of long-term losartan versus propranolol in lowering portal pressure in cirrhosis. AB - BACKGROUND & AIMS: It has been suggested that losartan, an angiotensin II (A-II) type 1 receptor blocker, may have a pronounced portal pressure reducing effect, far greater than that of propranolol. This randomized controlled trial compared the hemodynamic and renal effects of continued 6-week administration of losartan (n = 25) vs. propranolol (n = 15) in portal hypertensive patients with cirrhosis treated endoscopically after a variceal bleeding episode. METHODS: Hepatic venous pressure gradient (HVPG), systemic hemodynamics, renal function, and vasoactive factors were measured before and at 6 weeks of treatment. RESULTS: Losartan did not reduce HVPG (-2% +/- 12%, NS) but significantly decreased mean arterial pressure (MAP, -8% +/- 10%, P = 0.001). On the contrary, propranolol significantly reduced HVPG (-10% +/- 11%, P = 0.003) and cardiac output (-16% +/- 12%, P = 0.001) but did not modify MAP (2.5% +/- 10%, NS). Losartan increased A II levels, reduced aldosterone, and decreased glomerular filtration rate (GFR) in Child B patients. Propranolol did not modify renal function. Adverse events related to therapy were mild and similar in both groups. CONCLUSIONS: Unlike propranolol, long-term losartan administration does not significantly reduce HVPG in patients with cirrhosis treated after a variceal bleeding episode, and it caused hypotension and reduced GFR in patients with moderate liver failure. Therefore, losartan is not an alternative to propranolol in preventing variceal rebleeding. PMID- 11487548 TI - Hemodynamic effects of the angiotensin II receptor antagonist irbesartan in patients with cirrhosis and portal hypertension. AB - BACKGROUND & AIMS: Angiotensin II receptor antagonists have been proposed as new drugs for portal hypertension. This randomized, placebo-controlled, double-blind study aimed to assess the effect of the angiotensin II receptor antagonist irbesartan on portal and systemic hemodynamics and renal function in patients with cirrhosis. METHODS: Thirty-six patients with cirrhosis and portal hypertension received 150 mg/d irbesartan or placebo for 1 week. Systemic hemodynamics, kidney and liver function parameters were recorded regularly; hepatic venous pressure gradient and plasma renin were assessed on days 0 and 7. RESULTS: Irbesartan reduced the hepatic venous pressure gradient by 12.2% +/- 6.6% (P < 0.05) and mean arterial pressure by 5.3% +/- 4.0% in 13 of 18 verum patients. In 4 (22%) verum patients, arterial hypotension, accompanied by significant renal impairment, required withdrawal of irbesartan. In these patients, baseline plasma renin (P < 0.002) and cystatin C (P < 0.001) levels were higher, and creatinine clearance (P < 0.02), serum sodium (P < 0.01), and albumin (P < 0.05) were lower than in patients who tolerated irbesartan. Four of five patients with baseline renin >900 microU/mL developed treatment-limiting hypotension. CONCLUSIONS: The angiotensin II receptor antagonist irbesartan is not advisable in patients with advanced cirrhosis and high plasma renin because it may induce arterial hypotension and only moderately reduces portal pressure. PMID- 11487549 TI - Bile duct epithelia regulate biliary bicarbonate excretion in normal rat liver. AB - BACKGROUND & AIMS: A number of transporters and channels have been identified in cholangiocytes, but the role that bile ducts play in the formation of bile in vivo is unclear. We determined the contribution of cholangiocytes to bile flow and biliary bicarbonate excretion in normal rat liver. METHODS: Bile flow and biliary bicarbonate were measured in isolated rat livers perfused via both the portal vein and the hepatic artery because the hepatic artery provides the blood supply to bile ducts. Livers were perfused with secretin or acetylcholine (ACh), which respectively increase either adenosine 3',5'-cyclic monophosphate (cAMP) or cytosolic Ca(2+) in cholangiocytes. Livers also were perfused with glucagon or vasopressin to instead increase cAMP or cytosolic Ca(2+) in hepatocytes. RESULTS: Secretin increased biliary bicarbonate in a dose-dependent fashion and was much more effective when administered via the hepatic artery. Secretin did not affect bile flow. Similarly, ACh increased bicarbonate excretion when infused via the hepatic artery but not the portal vein. The effects of secretin were augmented by ACh, and this was prevented by cyclosporin A. The effects of ACh were blocked by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), 5-nitro2-(3 phenylpropylamino)benzoic acid (NPPB), or diphenylamine-2-carboxylic acid (DPC), and the effects of secretin were inhibited by NPPB or DPC and unaffected by DIDS. Neither glucagon nor vasopressin altered biliary bicarbonate. CONCLUSIONS: Biliary bicarbonate is regulated by cholangiocytes rather than hepatocytes in normal rat liver. ACh-induced bicarbonate excretion depends on both chloride channels and bicarbonate exchange, whereas secretin-induced bicarbonate excretion is independent of bicarbonate exchange. PMID- 11487550 TI - Tauroursodesoxycholate-induced choleresis involves p38(MAPK) activation and translocation of the bile salt export pump in rats. AB - BACKGROUND & AIMS: Canalicular secretion of bile acids is stimulated by tauroursodesoxycholate (TUDC). This study investigates the underlying mechanisms. METHODS: TUDC effects on mitogen-activated protein (MAP) kinases, taurocholate (TC) excretion, proteolysis, and the localization of the bile salt export pump (Bsep) were studied in rat hepatocytes and perfused liver. RESULTS: TUDC induced a transient and concentration-dependent activation of p38(MAPK) and of extracellular signal-regulated kinase 2 (Erk-2), but not of c-Jun-N-terminal kinase (JNK). In perfused liver, TUDC concentrations of 20 micromol/L was sufficient to elicit the MAP kinase responses and TC choleresis. SB 202190, a specific inhibitor of p38(MAPK), had no effect on TUDC- induced Erk activation but abolished the stimulatory effect of TUDC on TC excretion in perfused liver, indicating the requirement of p38(MAPK) in addition to the reported Erk dependence for the choleretic response. TUDC-induced stimulation of TC excretion was accompanied by a p38(MAPK)-dependent insertion of subcanalicular immunoreactive Bsep into the canalicular membrane. In addition TUDC induced a p38(MAPK)-sensitive inhibition of proteolysis. CONCLUSIONS: TUDC-induced stimulation of canalicular TC excretion involves a MAP kinase-dependent translocation of subcanalicular Bsep to the canalicular membrane. Dual activation of Erks and p38(MAPK) is required for the choleretic effect of both TUDC and hypo osmotic cell swelling. PMID- 11487551 TI - Dysphagia in a patient with lateral medullary syndrome: insight into the central control of swallowing. AB - BACKGROUND & AIMS: Central control of swallowing is regulated by a central pattern generator (CPG) positioned dorsally in the solitary tract nucleus and neighboring medullary reticular formation. The CPG serially activates the cranial nerve motor neurons, including the nucleus ambiguus and vagal dorsal motor nucleus, which then innervate the muscles of deglutition. This case provides insight into the central control of swallowing. METHODS: A 65-year-old man with a right superior lateral medullary syndrome presented with a constellation of symptoms, including dysphagia. The swallow was characterized using videofluoroscopy and esophageal motility and the results were compared with magnetic resonance imaging (MRI) findings. RESULTS: Videofluoroscopy showed intact lingual propulsion and volitional movements of the larynx. Distal pharyngeal peristalsis was absent, and the bolus did not pass the upper esophageal sphincter. Manometry showed proximal pharyngeal contraction and normal peristaltic activity in the lower esophagus (smooth muscle), but motor activity of the upper esophageal sphincter and proximal esophagus (striated muscle) was absent. MRI showed a lesion of the dorsal medulla. CONCLUSIONS: These findings are compatible with a specific lesion of the connections from a programming CPG in the solitary tract nucleus to nucleus ambiguus neurons, which supply the distal pharynx, upper esophageal sphincter, and proximal esophagus. There is functional preservation of the CPG control center in the solitary tract nucleus and of the vagal dorsal motor nucleus neurons innervating the smooth muscle esophagus. PMID- 11487552 TI - Loss of interstitial cells of cajal and inhibitory innervation in insulin dependent diabetes. AB - BACKGROUND & AIMS: Gastrointestinal complications of long-standing diabetes include nausea, vomiting, abdominal pain, diarrhea, and constipation. The pathophysiology underlying these symptoms is poorly understood. Recent evidence suggests an important role for interstitial cells of Cajal in controlling gastrointestinal motility. The aim of this study was to determine changes in interstitial cells of Cajal and enteric innervation in a patient with insulin dependent diabetes. METHODS: A full thickness jejunal biopsy was obtained from a 38-year-old insulin-dependent diabetic with evidence for diabetic gastroenteropathy. Immunohistochemistry, confocal microscopy, and 3-dimensional reconstruction techniques were used to quantify changes in the volume of interstitial cells of Cajal and enteric innervation. RESULTS: Interstitial cells of Cajal were markedly decreased throughout the entire thickness of the jejunum. A decrease in neuronal nitric oxide synthase, vasoactive intestinal peptide, PACAP, and tyrosine hydroxylase immunopositive nerve fibers was observed in circular muscle layer while substance P immunoreactivity was increased. CONCLUSIONS: The data suggest that long-standing diabetes is associated with a decrease in interstitial cells of Cajal volume and a decrease in inhibitory innervation, associated with an increase in excitatory innervation. The changes in interstitial cells of Cajal volume and enteric nerves may underlie the pathophysiology of gastrointestinal complications associated with diabetes and suggest future therapeutic targets. PMID- 11487553 TI - Numerous colonic adenomas in an individual with Bloom's syndrome. AB - Bloom's syndrome (BS) is a rare recessive disorder caused by germline mutation of the BLM gene. Individuals with BS manifest growth retardation, immunodeficiency, and a predisposition to cancer. In this report, we describe an individual with BS and multiple colonic adenomas reminiscent of familial adenomatous polyposis coli (FAP). Molecular studies revealed APC mutations in 4 of 6 adenomas, including 2 adenomas with the identical APC mutation and microsatellite instability in 1 of 6 adenomas. These results demonstrate similar pathways to colorectal neoplasia in BS as in the normal population and suggest that individuals with BS may be particularly susceptible to colorectal neoplasia. PMID- 11487554 TI - The American Gastroenterological Association standards for office-based gastrointestinal endoscopy services. AB - The American Gastroenterological Association (AGA) standards for office-based gastrointestinal endoscopy were written in response to market changes in physician reimbursements for many endoscopic procedures that will continue to drive their performance into unregulated physician offices. The AGA believes that patient safety is best protected if these standards are adopted by sites that also comply with state/federal laws for licensure or are certified as an ASC and/or are accredited by a nationally recognized accreditation program (e.g., the Joint Commission on Accreditation of Healthcare Organization's [JCAHO] new Office Based Surgery Standards). Heretofore, relevant practice standards for the performance of endoscopic procedures in these settings have not been available, a situation that the AGA believes puts patients at risk. These standards have been developed to reduce that risk. PMID- 11487555 TI - List of available training programs. PMID- 11487556 TI - Helicobacter pylori acquisition and transmission: where does it all begin? PMID- 11487558 TI - Angiotensin receptor blockade and portal hypertension: paradise gained and paradise lost. PMID- 11487557 TI - Morphogenesis and maintenance of the gastric epithelium: a role for sonic hedgehog? PMID- 11487559 TI - Ursodiol prevents UC-associated CRC. PMID- 11487560 TI - Persistent colonic inflammation in postinfectious IBS: symptom cause or curiosity? PMID- 11487561 TI - Chest pain and serotonin: a possible link? PMID- 11487562 TI - Molecular dissection of the mechanisms through which enteropathogenic Escherichia coli induce diarrhea. PMID- 11487563 TI - Shortcoming in the diagnosis of TPMT deficiency in a patient with Crohn's disease using phenotyping only. PMID- 11487564 TI - Excess cone cell proliferation due to lack of a functional NR2E3 causes retinal dysplasia and degeneration in rd7/rd7 mice. AB - The rd7 mouse is a model for hereditary retinal degeneration characterized clinically by retinal spotting throughout the fundus and late onset retinal degeneration, and histologically by retinal dysplasia manifesting as folds and whorls in the photoreceptor layer. This study demonstrates that the rd7 phenotype results from a splicing error created by a genomic deletion of an intron and part of an exon. Hematoxylin/eosin staining of rd7 tissue shows that the whorls in the outer nuclear layer of the retina do not appear during embryonic development but manifest by postnatal day 12.5 (P12.5). Furthermore, in situ hybridization data indicates that the Nr2e3 message is first present at barely discernable levels at embryonic day 18.5, becomes abundant by P2.5, and reaches maximal adult levels by P10.5. Results from these experiments indicate that Nr2e3 message is expressed prior to the development of S-cones. This data coincides with studies in humans showing that mutations in Nr2e3 result in a unique type of retinal degeneration known as enhanced S-cone syndrome, where patients have a 30-fold increase in S cone sensitivity compared to normal. Immunohistochemical staining of cone cells demonstrates that rd7 retinas have an increased number of cone cells compared to wild-type retinas. Thus, Nr2e3 may function by regulating genes involved in cone cell proliferation, and mutations in this gene lead to retinal dysplasia and degeneration by disrupting normal photoreceptor cell topography as well as cell cell interactions. PMID- 11487565 TI - Peptide-N-glycanases and DNA repair proteins, Xp-C/Rad4, are, respectively, active and inactivated enzymes sharing a common transglutaminase fold. AB - Yeast RAD4, its human ortholog Xp-C and their orthologs in other eukaryotes are DNA repair proteins which participate in nucleotide excision repair through a ubiquitin-dependent process. However, no conserved globular domains that might have shed light on their origin or functions have been reported for these proteins. By using sequence profile analysis, we show that RAD4/Xp-C proteins contain the ancient transglutaminase fold and are specifically related to the recently characterized peptide-N-glycanases (PNGases) which remove glycans from glycoproteins during their degradation. The PNGases retain the catalytic triad that is typical of this fold and are predicted to have a reaction mechanism similar to that involved in transglutamination. In contrast, the RAD4/Xp-C proteins are predicted to be inactive and are likely to only possess the protein interaction function in DNA repair. These proteins also contain a long, low complexity insert in the globular transglutaminase domain. The RAD4/Xp-C proteins, along with other inactive transglutaminase-fold proteins, represent a case of functional re-assignment of an ancient domain following the loss of the ancestral enzymatic activity. PMID- 11487566 TI - Functional analyses of two newly identified PITX2 mutants reveal a novel molecular mechanism for Axenfeld-Rieger syndrome. AB - The specific role of PITX2 in the pathogenesis of anterior segment dysgenesis has yet to be clearly defined. We provide here new insight into PITX2 pathogenesis through mutational and functional analyses. Three PITX2 mutations were found in a screen of 38 unrelated individuals affected with anterior segment anomalies (8%). All three mutations were found among the 21 individuals affected with Axenfeld Rieger syndrome (ARS). We have identified two novel mutations, a valine-->leucine (V45L) missense mutation at position 45 within the PITX2 homeodomain, and a seven amino acid duplication (7aaDup) of residues 6-12 of the homeodomain. DNA-binding studies of the two mutant PITX2 proteins demonstrated a <10-fold reduction in the DNA-binding activity of the V45L mutant, and a >100-fold reduction in activity of the 7aaDup mutant. Luciferase reporter assays showed a >200% increase in PITX2 transactivation activity of the V45L mutant, while the 7aaDup mutant was unable to transactivate at detectable levels. Our analyses of the V45L PITX2 mutant reveal that the DNA-binding domain of PITX2 can influence transactivation activity independently of DNA binding. Furthermore, our findings expand the hypothesis that the amount of residual PITX2 activity underlies the variable severity of ocular phenotypes that result from PITX2 mutation. For the first time, we present evidence that increased PITX2 activity may underlie the severe ARS ocular phenotype. We conclude that increased activity of one PITX2 allele may be as physiologically disruptive as a mutation that nullifies a PITX2 allele, with either condition resulting in ARS. PMID- 11487567 TI - Enzyme therapy for lysosomal acid lipase deficiency in the mouse. AB - Lysosomal acid lipase (LAL) is the critical enzyme for the hydrolysis of the triglycerides (TG) and cholesteryl esters (CE) delivered to lysosomes. Its deficiency produces two human phenotypes, Wolman disease (WD) and cholesteryl ester storage disease (CESD). A targeted disruption of the LAL locus produced a null (lal( -/-)) mouse model that mimics human WD/CESD. The potential for enzyme therapy was tested using mannose terminated human LAL expressed in Pichia pastoris (phLAL), purified, and administered by tail vein injections to lal( -/-) mice. Mannose receptor (MR)-dependent uptake and lysosomal targeting of phLAL were evidenced ex vivo using competitive assays with MR-positive J774E cells, a murine monocyte/macrophage line, immunofluorescence and western blots. Following (bolus) IV injection, phLAL was detected in Kupffer cells, lung macrophages and intestinal macrophages in lal( -/-) mice. Two-month-old lal( -/-) mice received phLAL (1.5 U/dose) or saline injections once every 3 days for 30 days (10 doses). The treated lal( -/-) mice showed nearly complete resolution of hepatic yellow coloration; hepatic weight decreased by approximately 36% compared to PBS-treated lal( -/-) mice. Histologic analyses of numerous tissues from phLAL-treated mice showed reductions in macrophage lipid storage. TG and cholesterol levels decreased by approximately 50% in liver, 69% in spleen and 50% in small intestine. These studies provide feasibility for LAL enzyme therapy in human WD and CESD. PMID- 11487568 TI - The importance of gene dosage studies: mutational analysis of the parkin gene in early-onset parkinsonism. AB - Early-onset parkinsonism (EOP) may be associated with different mutations in the parkin gene, including exon deletions and duplications. To test for gene dosage alterations, we developed a new method of quantitative duplex PCR using the fluorescence resonance energy transfer technique on the LightCycler (Roche Diagnostics). In 21 patients with EOP, three mutations (a single base pair substitution in exon 3 and small deletions in exon 9) were detected by conventional mutational screening (single-strand conformation polymorphism and sequence analysis), while alterations of gene dosage were found in seven patients. We identified heterozygous and compound heterozygous deletions of exons 2, 3, 5 and 7. The latter was also found in the homozygous state. In addition, two heterozygous duplications of exon 4 were observed. Remarkably, two patients carried more than two parkin mutations. This is the first study systematically screening all 12 exons of parkin by real-time, kinetic quantification and clearly shows that mutational analysis of the parkin gene should include gene dosage studies. Furthermore, our method of quantitative PCR is easily applicable to any other gene to be screened for deletions or duplications of whole exons. PMID- 11487569 TI - Mismatch repair detection (MRD): high-throughput scanning for DNA variations. AB - Although there are several methods for genotyping previously identified single nucleotide polymorphisms (SNPs), there is a paucity of approaches for high throughput scanning for unknown variations. Mismatch repair detection (MRD) utilizes a bacterial mismatch repair system in vivo to detect sequence variants in human DNA samples. We describe modifications in MRD that allow a high degree of parallel processing, and use this modified version to accurately scan for variations in 35 different human DNA fragments simultaneously. MRD's potential for high-throughput scanning can be used to identify new SNPs and to comprehensively compare sequences between patients and controls for identifying disease susceptibility alleles. PMID- 11487570 TI - Pathogenic APP mutations near the gamma-secretase cleavage site differentially affect Abeta secretion and APP C-terminal fragment stability. AB - Release of amyloid beta (Abeta) from the amyloid precursor protein (APP) requires cleavages by beta- and gamma-secretases and plays a crucial role in Alzheimer's disease (AD) pathogenesis. Missense mutations in the APP gene causing familial AD are clustered around the beta-, alpha- and particular gamma-secretase cleavage sites. We systematically compare in primary neurons the effect on APP processing of a series of clinical APP mutations (two of which not characterized before) located in close proximity to the gamma-secretase cleavage site. We confirm and extend previous observations showing that all these mutations (T714I, V715M, V715A, I716V, V717I and V717L) affect gamma-secretase cleavage causing an increased relative ratio of Abeta42 to Abeta40. Taking advantage of these extended series of APP mutations we were able to demonstrate an inverse correlation between these ratios and the age at onset of the disease in the different families. In addition, a subset of mutations caused the accumulation of APP C-terminal fragments indicating that these mutations also influence the stability of APP C-terminal fragments. However, it is unlikely that these fragments contribute significantly to the disease process. PMID- 11487571 TI - Mutation of the gene encoding the enamel-specific protein, enamelin, causes autosomal-dominant amelogenesis imperfecta. AB - Amelogenesis imperfecta (AI) is a group of inherited defects of dental enamel formation that shows both clinical and genetic heterogeneity. To date, mutations in the gene encoding amelogenin have been shown to underlie a subset of the X linked recessive forms of AI. Although none of the genes underlying autosomal dominant or autosomal-recessive AI have been identified, a locus for a local hypoplastic form has been mapped to human chromosome 4q11-q21. In the current investigation, we have analysed a family with an autosomal-dominant, smooth hypoplastic form of AI. Our results have shown that a splicing mutation in the splice donor site of intron 7 of the gene encoding the enamel-specific protein enamelin underlies the phenotype observed in this family. This is the first autosomal-dominant form of AI for which the genetic mutation has been identified. As this type of AI is clinically distinct from that localized previously to chromosome 4q11-q21, these findings highlight the need for a molecular classification of this group of disorders. PMID- 11487572 TI - SCA7 mouse models show selective stabilization of mutant ataxin-7 and similar cellular responses in different neuronal cell types. AB - Accumulation of expanded polyglutamine proteins and selective pattern of neuronal loss are hallmarks of at least eight neurodegenerative disorders, including spinocerebellar ataxia type 7 (SCA7). We previously described SCA7 mice displaying neurodegeneration with progressive ataxin-7 accumulation in two cell types affected in the human pathology. We describe here a new transgenic model with a more widespread expression of mutant ataxin-7, including neuronal cell types unaffected in SCA7. In these mice a similar handling of mutant ataxin-7, including a cytoplasm to nucleus translocation and accumulation of N-terminal fragments, was observed in all neuronal populations studied. An extensive screen for chaperones, proteasomal subunits and transcription factors sequestered in nuclear inclusions (NIs) disclosed no pattern unique to neurons undergoing degeneration in SCA7. In particular, we found that the mouse TAF(II)30 subunit of the TFIID initiation complex is markedly accumulated in NIs, even though this protein does not contain a polyglutamine stretch. A striking discrepancy between mRNA and ataxin-7 levels in transgenic mice expressing the wild-type protein but not in those expressing the mutant one, indicates a selective stabilization of mutant ataxin-7, both in this model and the P7E/N model described previously. These mice therefore provide in vivo evidence that the polyglutamine expansion mutation can stabilize its target protein. PMID- 11487573 TI - Instability of a (CGG)98 repeat in the Fmr1 promoter. AB - Fragile X syndrome is one of 14 trinucleotide repeat diseases. It arises due to expansion of a CGG repeat which is present in the 5'-untranslated region of the FMR1 gene, disruption of which leads to mental retardation. The mechanisms involved in trinucleotide repeat expansion are poorly understood and to date, transgenic mouse models containing transgenic expanded CGG repeats have failed to reproduce the instability seen in humans. As both cis-acting factors and the genomic context of the CGG repeat are thought to play a role in expansion, we have now generated a knock-in mouse Fmr1 gene in which the murine (CGG)8 repeat has been exchanged with a human (CGG)98 repeat. Unlike other CGG transgenic models, this model shows moderate CGG repeat instability upon both in maternal and paternal transmission. This model will now enable us to study the timing and the mechanism of repeat expansion in mice. PMID- 11487574 TI - The melanocortin-1-receptor gene is the major freckle gene. AB - Ephelides and solar lentigines are different types of pigmented skin lesions. Ephelides appear early in childhood and are associated with fair skin type and red hair. Solar lentigines appear with increasing age and are a sign of photodamage. Both lesions are strong risk indicators for melanoma and non melanoma skin cancer. Melanocortin-1-receptor (MC1R) gene variants are also associated with fair skin, red hair and melanoma and non-melanoma skin cancer. The purpose of this study was to investigate the relationship between MC1R gene variants, ephelides and solar lentigines. In a large case-control study, patients with melanoma and non-melanoma skin cancer and subjects without a history of skin cancer were studied. In all participants, the presence of ephelides in childhood and solar lentigines by physical examination was assessed according to strict definitions. The entire coding sequence of the MC1R gene was analyzed by single strand conformation polymorphism analysis followed by sequence analyses. Carriers of one or two MC1R gene variants had a 3- and 11-fold increased risk of developing ephelides, respectively (both P < 0.0001), whereas the risk of developing severe solar lentigines was increased 1.5- and 2-fold (P = 0.035 and P < 0.0001), respectively. These associations were independent of skin type and hair color, and were comparable in patients with and without a history of skin cancer. The population attributable risk for ephelides to MC1R gene variants was 60%, i.e. 60% of the ephelides in the population was caused by MC1R gene variants. A dosage effect was found between the degree of ephelides and the number of MC1R gene variants. As nearly all individuals with ephelides were carriers of at least one MC1R gene variant, our data suggest that MC1R gene variants are necessary to develop ephelides. The results of the study also suggest that MC1R gene variants play a role, although less important, in the development of solar lentigines. PMID- 11487575 TI - Mutations in the novel protocadherin PCDH15 cause Usher syndrome type 1F. AB - We have determined the molecular basis for Usher syndrome type 1F (USH1F) in two families segregating for this type of syndromic deafness. By fluorescence in situ hybridization, we placed the human homolog of the mouse protocadherin Pcdh15 in the linkage interval defined by the USH1F locus. We determined the genomic structure of this novel protocadherin, and found a single-base deletion in exon 10 in one USH1F family and a nonsense mutation in exon 2 in the second. Consistent with the phenotypes observed in these families, we demonstrated expression of PCDH15 in the retina and cochlea by RT-PCR and immunohistochemistry. This report shows that protocadherins are essential for maintenance of normal retinal and cochlear function. PMID- 11487576 TI - AT-rich islands in genomic DNA as a novel target for AT-specific DNA-reactive antitumor drugs. AB - Interstrand cross-links at T(A/T)4A sites in cellular DNA are associated with hypercytotoxicity of an anticancer drug, bizelesin. Here we evaluated whether these lethal effects reflect targeting critical genomic regions. An in silico analysis of human sequences showed that T(A/T)4A motifs are on average scarce and scattered. However, significantly higher local motif densities were identified in distinct minisatellite regions (200-1000 base pairs of approximately 85-100% AT), herein referred to as "AT islands." Experimentally detected bizelesin lesions agree with these in silico predictions. Actual bizelesin adducts clustered within the model AT island naked DNA, whereas motif-poor sequences were only sparsely adducted. In cancer cells, bizelesin produced high levels of lesions (approximately 4.7-7.1 lesions/kilobase pair/microM drug) in several prominent AT islands, compared with markedly lower lesion levels in several motif-poor loci and in bulk cellular DNA (approximately 0.8-1.3 and approximately 0.9 lesions/kilobase pair/microM drug, respectively). The identified AT islands exhibit sequence attributes of matrix attachment regions (MARs), domains that organize DNA loops on the nuclear matrix. The computed "MAR potential" and propensity for supercoiling-induced duplex destabilization (both predictive of strong MARs) correlate with the total number of bizelesin binding sites. Hence, MAR-like AT-rich non-coding domains can be regarded as a novel class of critical targets for anticancer drugs. PMID- 11487577 TI - Induction of the transcriptional repressor Yin Yang-1 by vascular cell injury. Autocrine/paracrine role of endogenous fibroblast growth factor-2. AB - Yin Yang-1 (YY1) is a multifunctional transcription factor that can repress the expression of many growth factor, hormone, and cytokine genes implicated in atherogenesis. YY1 expression is activated in rat vascular smooth muscle cells shortly after injury. YY1 DNA binding activity paralleled elevated protein levels in the nucleus. Smooth muscle cell injury triggered the rapid extracellular release of immunoreactive fibroblast growth factor-2 (FGF-2). YY1 induction after injury was blocked by neutralizing antibodies directed against FGF-2. This growth factor increased YY1 mRNA and protein expression and stimulated YY1 binding and transcriptional activity. Overexpression of YY1 inhibited smooth muscle cell replication. Immunohistochemical analysis demonstrated YY1 staining in medial smooth muscle cells, coincident with FGF-2 expression. Proliferating cell nuclear antigen staining, in contrast, was confined mainly to the atherosclerotic intima. This is the first demonstration that YY1 is induced by either injury or FGF-2, is differentially expressed in normal and diseased human arteries, and that its overexpression inhibits vascular smooth muscle but not endothelial cell replication. PMID- 11487579 TI - Fast coordination changes in cytochrome c do not necessarily imply folding. AB - Folding of globular proteins occurs with rates that range from microseconds to minutes; consequently, it has been necessary to develop new strategies to follow the faster processes that exceed stopped-flow capabilities. Rapid photochemical methods have been employed to study the rate of folding of reduced cytochrome c. In this protein, the iron of the covalently bound heme binds a His and a Met, proximal and distal. Unfolding by guanidine or urea weakens the Fe-Met bond, and the reduced unfolded cytochrome c easily binds CO and other heme ligands, which would react slowly or not at all with the native protein. Therefore in the presence of CO, reduced cytochrome c unfolds at lower denaturant concentrations than in the absence of this ligand, and rapid photochemical removal of CO from unfolded cytochrome c, is expected to trigger at least an incomplete refolding. This approach is complicated by the breakage of the proximal His-Fe bond that may occur as a consequence of CO photodissociation in the unfolded cytochrome c because of the so-called base elimination mechanism. Rebinding of CO to the four coordinate heme yields kinetic intermediates unrelated to folding. Our hypothesis is supported by parallel observations carried out with protoheme and microperoxidase. PMID- 11487578 TI - Axin-dependent phosphorylation of the adenomatous polyposis coli protein mediated by casein kinase 1epsilon. AB - Axin and the adenomatous polyposis coli protein (APC) interact to down-regulate the proto-oncogene beta-catenin. We show that transposition of an axin-binding site can confer beta-catenin regulatory activity to a fragment of APC normally lacking this activity. The fragment containing the axin-binding site also underwent hyperphosphorylation when coexpressed with axin. The phosphorylation did not require glycogen synthase kinase 3beta but instead required casein kinase 1epsilon, which bound directly to axin. Mutation of conserved serine residues in the beta-catenin regulatory motifs of APC interfered with both axin-dependent phosphorylation and phosphorylation by CKIepsilon and impaired the ability of APC to regulate beta-catenin. These results suggest that the axin-dependent phosphorylation of APC is mediated in part by CKIepsilon and is involved in the regulation of APC function. PMID- 11487580 TI - Ligand-switching intermediates for the CO-sensing transcriptional activator CooA measured by pulse radiolysis. AB - CooA is a heme-containing and CO-sensing transcriptional activator whose activity is regulated by CO. The protoheme that acts as a CO sensor in CooA shows unique properties for its coordination structure. The Cys75 axial ligand of the ferric heme is replaced by His77 upon the reduction of the heme iron and vice versa. In this work, the ligand-switching process induced by the reduction of the heme was investigated by the technique of pulse radiolysis. Hydrated electron reduced the heme iron in ferric CooA within 1 micros to form the first intermediate with the Soret peak at 440 nm, suggesting that a six-coordinate ferrous heme with a thiolate axial ligand was formed initially. The first intermediate was converted into the second intermediate with the time constant of 40 micros (k = 2.5 x 10(4) x s(-1)). In the second intermediate, the thiolate from Cys75 was thought to be protonated and/or the Fe-S bond was thought to be elongated. The second intermediate was converted into the final reduced form with the time constant of 2.9 ms (k = 3.5 x 10(2) x s(-1)) for wild-type CooA. The ligand exchange between Cys75 and His77 took place during the conversion of the second intermediate into the final reduced form. PMID- 11487581 TI - Indecisive M13 procoat protein mutants bind to SecA but do not activate the translocation ATPase. AB - The M13 procoat protein serves as the paradigm for the Sec-independent membrane insertion pathway. This protein is inserted into the inner membrane of Escherichia coli with two hydrophobic regions and a central periplasmic loop region of 20 amino acid residues. Extension of the periplasmic loop region renders M13 procoat membrane insertion Sec-dependent. Loop regions with 118 or more residues required SecA and SecYEG and were efficiently translocated in vivo. Two mutants having loop regions of 80 and 100 residues, respectively, interacted with SecA but failed to activate the membrane translocation ATPase of SecA in vitro. Similarly, a procoat mutant with two additional glutamyl residues in the loop region showed binding to SecA but did not stimulate the ATPase. The three mutants were also defective for precursor-stimulated binding of SecA to the membrane surface. Remarkably, the mutant proteins act as competitive inhibitors of the Sec translocase. This suggests that the region to be translocated is sensed by SecA but the activation of the SecA translocation ATPase is only successful for substrates with a minimum length of the translocated region. PMID- 11487583 TI - Antiviral activity and structural characteristics of the nonglycosylated central subdomain of human respiratory syncytial virus attachment (G) glycoprotein. AB - Segments of the cystine noose-containing nonglycosylated central subdomain, residues 149-197, of the attachment (G) glycoprotein of human respiratory syncytial virus (HRSV) have been assessed for impact on the cytopathic effect (CPE) of respiratory syncytial virus (RSV). Nalpha-acetyl residues 149-197-amide (G149-197), G149-189, and G149-177 of the A2 strain of HRSV protected 50% of human epithelial HEp-2 cells from the CPE of the A2 strain at concentrations (IC(50)) between 5 and 80 microm. Cystine noose-containing peptides G171-197 and G173-197 did not inhibit the CPE even at concentrations above 150 microm. Systematic C- and N-terminal truncations from G149-189 and G149-177 and alanine substitutions within G154-177 demonstrated that residues 166-170 (EVFNF), within a sequence that is conserved in HRSV strains, were critical for inhibition. Concordantly, G154-177 of bovine RSV and of an antibody escape mutant of HRSV with residues 166-170 of QTLPY and EVSNP, respectively, were not inhibitory. Surprisingly, a variant of G154-177 with an E166A substitution had an IC(50) of 750 nm. NMR analysis demonstrated that G149-177 adopted a well-defined conformation in solution, clustered around F168 and F170. G154-170, particularly EVFNF, may be important in binding of RSV to host cells. These findings constitute a promising platform for the development of antiviral agents for RSV. PMID- 11487582 TI - The polymorphism at codon 54 of the FABP2 gene increases fat absorption in human intestinal explants. AB - Based on titration microcalorimetry and Caco-2 cell line transfection studies, it has been suggested that the A54T of the FABP2 gene plays a significant role in the assimilation of dietary fatty acids. However, reports were divergent with regard to the in vivo interaction between this polymorphism and postprandial lipemia. We therefore determined the influence of this intestinal fatty acid binding protein polymorphism on intestinal fat transport using the human jejunal organ culture model, thus avoiding the interference of various circulating factors capable of metabolizing in vivo postprandial lipids. Analysis of DNA samples from 32 fetal intestines revealed 22 homozygotes for the wild-type Ala 54/Ala-54 genotype (0.83) and 10 heterozygotes for the polymorphic Thr-54/Ala-54 genotype (0.17). The Thr-encoding allele was associated with increased secretion of newly esterified triglycerides, augmented de novo apolipoprotein B synthesis, and elevated chylomicron output. On the other hand, no alterations were found in very low density lipoprotein and high density lipoprotein production, apolipoprotein A-I biogenesis, or microsomal triglyceride transfer protein mass and activity. Similarly, the alanine to threonine substitution at residue 54 did not result in changes in brush border hydrolytic activities (sucrase, glucoamylase, lactase, and alkaline phosphatase) or in glucose uptake or oxidation. Our data clearly document that the A54T polymorphism of FABP2 specifically influences small intestinal lipid absorption without modifying glucose uptake or metabolism. It is proposed that, in the absence of confounding factors such as environmental and genetic variables, the FABP2 polymorphism has an important effect on postprandial lipids in vivo, potentially influencing plasma levels of lipids and atherogenesis. PMID- 11487584 TI - Transforming growth factor-alpha prevents detachment-induced inhibition of c-Src kinase activity, Bcl-XL down-regulation, and apoptosis of intestinal epithelial cells. AB - Detachment of epithelial cells from the extracellular matrix (ECM) results in apoptosis, a phenomenon often referred to as anoikis. Acquisition of anoikis resistance is now thought to be a prerequisite for the progression of carcinomas. Colorectal cancer cells frequently secrete epidermal growth factor receptor (EGFR) ligands, which are known to have anti-apoptotic activity. However, whether these ligands have the ability to inhibit anoikis of intestinal epithelial cells is unclear, since at least in some cell types efficient EGFR signaling requires cell-ECM adhesion. Here we report that transforming growth factor-alpha (TGF alpha), an EGFR ligand that is frequently secreted by colorectal cancer cells, strongly inhibits anoikis of the non-malignant rat intestinal epithelial cell lines, IEC-18 and RIE-1. TGF-alpha exerts its anti-anoikis effect by preventing detachment-induced inhibition of c-Src kinase activity. We also show that Fas activation, a molecular event known to play a critical role in anoikis, is not suppressed by TGF-alpha. On the other hand, this growth factor strongly inhibits the detachment-induced down-regulation of Bcl-X(L), another change that is involved in the induction of anoikis. We further demonstrate that this inhibition occurs in a c-Src-dependent manner. We conclude that TGF-alpha has the ability to suppress anoikis of intestinal epithelial cells, at least in part, by reverting the loss of c-Src activity and Bcl-X(L) expression induced by detachment from the ECM. PMID- 11487585 TI - Hematopoietic progenitor kinase 1 associates physically and functionally with the adaptor proteins B cell linker protein and SLP-76 in lymphocytes. AB - B cell linker protein (BLNK) is a SLP-76-related adaptor protein essential for signal transduction from the BCR. To identify components of BLNK-associated signaling pathways, we performed a phosphorylation-dependent yeast two-hybrid analysis using BLNK probes. Here we report that the serine/threonine kinase hematopoietic progenitor kinase 1 (HPK1), which is activated upon antigen receptor stimulation and which has been implicated in the regulation of MAP kinase pathways, interacts physically and functionally with BLNK in B cells and with SLP-76 in T cells. This interaction requires Tyr(379) of HPK1 and the Src homology 2 (SH2) domain of BLNK/SLP-76. Via homology modeling, we defined a consensus binding site within ligands for SLP family SH2 domains. We further demonstrate that the SH2 domain of SLP-76 participates in the regulation of AP-1 and NFAT activation in response to T cell receptor (TCR) stimulation and that HPK1 inhibits AP-1 activation in a manner partially dependent on its interaction with SLP-76. Our data are consistent with a model in which full activation of HPK1 requires its own phosphorylation on tyrosine and subsequent interaction with adaptors of the SLP family, providing a mechanistic basis for the integration of this kinase into antigen receptor signaling cascades. PMID- 11487586 TI - Agonist-dependent repression mediated by mutant estrogen receptor alpha that lacks the activation function 2 core domain. AB - Nuclear receptor corepressor (N-CoR) and silencing mediator of retinoid and thyroid hormone receptors (SMRT) form heterogeneous complexes with various histone deacetylases (HDACs). In this report, we found that ER alpha-Delta AF2, a mutant estrogen receptor alpha (ER alpha) deleted for the C-terminal activation function 2 (AF2) core domain, directs estradiol (E(2))-dependent repression and impairs E(2)-induced transactivation by wild type ER alpha. This repression required coexpressed BRG1 in SW-13 cells that lack BRG1, the ATPase constituent of the chromatin-remodeling SWI.SNF complex, and was abolished by HDAC inhibitor trichostatin A. We further demonstrated that ER alpha-Delta AF2 constitutively associates with SMRT but binds DNA in an E(2)-dependent manner in vivo. These results suggest that ER alpha-Delta AF2 and similar mutant receptors recently found associated with certain tumors may actively perturb the normal E(2) signaling via SWI/SNF, N-CoR/SMRT, and HDAC. PMID- 11487587 TI - Negative cooperativity of substrate binding but not enzyme activity in wild-type and mutant forms of CTP:glycerol-3-phosphate cytidylyltransferase. AB - CTP:glycerol-3-phosphate cytidylyltransferase (GCT) catalyzes the synthesis of CDP-glycerol for teichoic acid biosynthesis in certain Gram-positive bacteria. This enzyme is a model for a cytidylyltransferase family that includes the enzymes that synthesize CDP-choline and CDP-ethanolamine for phosphatidylcholine and phosphatidylethanolamine biosynthesis. We have used quenching of intrinsic tryptophan fluorescence to measure binding affinities of substrates to the GCT from Bacillus subtilis. Binding of either CTP or glycerol-3-phosphate to GCT was biphasic, with two binding constants of about 0.1-0.3 and 20-40 microm for each substrate. The stoichiometry of binding was 2 molecules of substrate/enzyme dimer, so the two binding constants represented distinctly different affinities of the enzyme for the first and second molecule of each substrate. The biphasic nature of binding was observed with the wild-type GCT as well as with several mutants with altered Km or kcat values. This negative cooperativity of binding was also seen when a catalytically defective mutant was saturated with two molecules of CTP and then titrated with glycerol-3-phosphate. Despite the pronounced negative cooperativity of substrate binding, negative cooperativity of enzyme activity was not observed. These data support a mechanism in which catalysis occurs only when the enzyme is fully loaded with 2 molecules of each substrate/enzyme dimer. PMID- 11487589 TI - A comprehensive view of regulation of gene expression by double-stranded RNA mediated cell signaling. AB - Double-stranded (ds) RNA, a common component of virus-infected cells, is a potent inducer of the type I interferon and other cellular genes. For identifying the full repertoire of human dsRNA-regulated genes, a cDNA microarray hybridization screening was conducted using mRNA from dsRNA-treated GRE cells. Because these cells lack all type I interferon genes, the possibility of gene induction by autocrine actions of interferon was eliminated. Our screen identified 175 dsRNA stimulated genes (DSG) and 95 dsRNA-repressed genes. A subset of the DSGs was also induced by different inflammatory cytokines and viruses demonstrating interconnections among disparate signaling pathways. Functionally, the DSGs encode proteins involved in signaling, apoptosis, RNA synthesis, protein synthesis and processing, cell metabolism, transport, and structure. Induction of such a diverse family of genes by dsRNA has major implications in host-virus interactions and in the use of RNA(i) technology for functional ablation of specific genes. PMID- 11487588 TI - Determinants involved in Kv1 potassium channel folding in the endoplasmic reticulum, glycosylation in the Golgi, and cell surface expression. AB - Kv1.1 and Kv1.4 potassium channels are expressed as mature glycosylated proteins in brain, whereas they exhibited striking differences in degree of trans-Golgi glycosylation conversion and high cell surface expression when they were transiently expressed as homomers in cell lines. Kv1.4 exhibited a 70% trans Golgi glycosylation conversion, whereas Kv1.1 showed none, and Kv1.4 exhibited a approximately 20-fold higher cell surface expression level as compared with Kv1.1. Chimeras between Kv1.4 and Kv1.1 and site-directed mutants were constructed to identify amino acid determinants that affected these processes. Truncating the cytoplasmic C terminus of Kv1.4 inhibited its trans-Golgi glycosylation and high cell surface expression (as shown by Li, D., Takimoto, K., and Levitan, E. S. (2000) J. Biol. Chem. 275, 11597-11602), whereas truncating this region on Kv1.1 did not affect either of these events, indicating that its C terminus is not a negative determinant for these processes. Exchanging the C terminus between these channels showed that there are other regions of the protein that exert a positive or negative effect on these processes. Chimeric constructs between Kv1.4 and Kv1.1 identified their outer pore regions as major positive and negative determinants, respectively, for both trans-Golgi glycosylation and cell surface expression. Site-directed mutagenesis identified a number of amino acids in the pore region that are involved in these processes. These data suggest that there are multiple positive and negative determinants on both Kv1.4 and Kv1.1 that affect channel folding, trans-Golgi glycosylation conversion, and cell surface expression. PMID- 11487590 TI - Response to "Interpretation, design, and analysis of gene array expression experiments". PMID- 11487591 TI - Age-specific demographic profiles of longevity mutants in Caenorhabditis elegans show segmental effects. AB - Demographic profiles of several single-gene longevity mutants of the nematode Caenorhabditis elegans reveal segmental (age-specific) effects on mortality. The mortality profiles of wild-type worms were examined across multiple replicate cultures containing 100,000 or more nematodes and found to be quite replicable, although clear environmental effects are routinely found. The combined profile of wild type was compared with those of three long-lived mutants to determine how age-specific mortality is altered by mutations in age-1, clk-1, or spe-26. In all four genotypes, death rates fit a two-stage Gompertz model better than a one stage Gompertz; that is, mortality levels off at later ages. The largest genetic effect on mortality was that of an age-1 mutation, which lowered mortality more than fivefold at most later ages. In contrast, a spe-26 mutant had a tenfold lower mortality until approximately 2 weeks of age but ultimately achieved a higher mortality, whereas clk-1 mutants show slightly higher mortality than wild type during the fertile period, early in life, but ultimately level off at lower mortality. Each mutant thus has a distinctive profile of age-specific mortalities that could suggest the time of action of each gene. PMID- 11487592 TI - Genes that prolong life: relationships of growth hormone and growth to aging and life span. AB - Mutant mice with a combined deficiency of growth hormone (GH), prolactin, and thyrotropin, and knockout mice with GH resistance, live longer than their normal siblings. The extension of life span in these animals is very large (up to 65%), reproducible, and not limited to any particular genetic background or husbandry conditions. In addition to demonstrating that genes control aging in mammals, these findings suggest that GH actions, growth, and body size may have important roles in the determination of life span. We describe the key phenotypic characteristics of long-living mutant and knockout mice, with an emphasis on those characteristics that may be related to delayed aging in these animals. We also address the broader topic of the relationship between GH, growth, maturation, body size, and aging, and we attempt to reconcile the well-publicized antiaging action of GH with the evidence that suppression of GH release or action can prolong life. PMID- 11487593 TI - Gene expression of cyclooxygenase in the aging heart. AB - Cyclooxygenase (COX) is the key rate-limiting enzyme in the prostaglandin synthetic pathway. Two isoforms of COX have been identified: a constitutive COX-1 and an inducible COX-2, which is activated in response to various stimuli. We investigated the changes of COX-1 and COX-2 in rat heart during aging. We measured the age-related changes in the mRNA and protein levels of COX by using reverse-transcription polymerase chain reaction and Western blotting, respectively. COX-2 mRNA and protein levels increased with age, whereas those of COX-1 showed no change. The COX activity determined by prostaglandin E(2) production increased with age. Because the COX-catalyzed arachidonate cascade is an important source of reactive oxygen species (ROS) generation, changes in ROS generation and lipid peroxidation were also assessed. The amount of ROS generated by the COX pathway increased with age, as did the total ROS generation and lipid peroxidation. These results show that COX-2 activity increases with age, partially because of elevated transcriptional expression and protein content, and they suggest that increased COX-2 can play a role in oxidative alterations in the aged heart. PMID- 11487594 TI - Differential involvement of conotoxin-sensitive mechanisms in neurogenic vasodilatation responses: effects of age. AB - During aging there is a decline in sensory nerve function that is associated with reduced neurogenic inflammation and poor wound repair. The cellular mechanism(s) responsible for this decline in function with age is not well understood. We previously reported that sensory nerves in aged rats release sensory neuropeptides preferentially in response to low-frequency (5 Hz) as compared with higher-frequency (15 Hz) antidromic electrical stimulation, and that low frequency transcutaneous electrical nerve stimulation accelerates wound healing. The present study investigates possible mechanisms for this preferential response. Using laser Doppler techniques, we have measured changes in blood flow in the base of vacuum-induced blisters induced in the rat hind footpad of young and old animals in response to low-frequency (5 Hz) or high-frequency (15 Hz) electrical stimulation (20 V, 2 ms for 1 minute) of the sciatic nerve. The relative contributions of the sensory neuropeptides, substance P and calcitonin gene-related peptide (CGRP), and of N-type voltage-gated calcium channels to the vascular responses were assessed by using the specific receptor antagonists RP67580, which is 2-(1-imino-2-(2 methoxy phyenyl) ethyl)-7,7 diphenyl-4 perhydroisoindolone-(3aR, 7aR); CGRP(8-37); and omega-conotoxin GVIA (Conus geographus), respectively. The results showed a greater involvement of substance P at high-frequency electrical stimulation and of CGRP at low-frequency stimulation. Our finding that omega-conotoxin-sensitive N-type calcium channel function was preserved with age and was only involved in the vascular response to low-frequency electrical stimulation could explain our previous report demonstrating beneficial effects of low-frequency transcutaneous electrical nerve stimulation to wound repair in aged animals. The current results have important practical implications for improving tissue repair in the aged. PMID- 11487595 TI - Growth hormone increases regional coronary blood flow and capillary density in aged rats. AB - In this study, we examined the effects of age and growth hormone replacement on both coronary blood flow and capillary density. Blood flow was measured by using [(14)C]-iodoantipyrine in three groups of anesthetized Brown Norway x Fischer 344 rats: young vehicle-treated animals (6 months; n = 13), old vehicle treated animals (30 months; n = 9), and old animals treated with bovine growth hormone (200 microg/kg) twice a day for 30 days (30 months; n = 7). Capillary density was measured by color segmentation analysis of sections stained for platelet endothelial cell adhesion molecule-1. In all regions examined, coronary blood flow decreased with age, and growth hormone administration resulted in an increase in flow compared to vehicle-treated animals. Capillary density decreased with age in the apex and the left ventricular middle segment. In response to growth hormone administration, capillary density increased significantly in the apex but not in other regions of the heart. Our results demonstrate that growth hormone enhances regional myocardial blood flow in the aged heart and suggest that part of this effect could be due to an increase in capillary density. PMID- 11487596 TI - Long-term use of low molecular weight heparin for venous thromboembolism prophylaxis in French geriatric settings. PMID- 11487598 TI - Chronic rheumatologic symptoms in a tri-ethnic sample of men and women aged 75 and older. AB - BACKGROUND: The high prevalence of musculoskeletal symptoms in elderly persons tends to obscure the recognition of specific medical conditions. The purpose of this study is to describe the prevalence of chronic rheumatologic symptoms and the associated measures of health status in an older population. METHODS: This is a cross-sectional study of 507 noninstitutionalized tri-ethnic men and women aged 75 and older living in Galveston County, Texas. A home interview collected data on demographics, chronic medical conditions, cognition, depression, and functional status. Site, severity, and duration of rheumatologic symptoms such as morning stiffness, body tenderness, and body aching in the past month were also collected. RESULTS: There was a high prevalence of nonspecific rheumatologic symptoms such as morning stiffness (32% in the shoulder girdle, 31% in the hip girdle), tenderness to touch (9%), and generalized body aching (11%) in the study population. Twenty-one percent of the subjects reported either bilateral shoulder or hip girdle stiffness or tenderness lasting more than 30 minutes almost every day or every day or generalized body aching most of the time during the past month. Age-, gender-, and ethnicity-adjusted multivariate analyses showed that more than three self-reported chronic medical conditions, poor or fair self reported health, impairment in instrumental activities of daily living (IADL), and the presence of depressive or anxiety symptoms were associated with the presence of these chronic rheumatologic symptoms. CONCLUSION: Chronic rheumatologic symptoms are common in people aged 75 and older. Such symptoms are markers for underlying poor health and for anxiety and depression among older subjects. PMID- 11487597 TI - Physical activity outcomes of CHAMPS II: a physical activity promotion program for older adults. AB - BACKGROUND: Despite well-known benefits of physical activity for older adults, about two thirds are underactive. Community-based programs are needed to facilitate increased physical activity. We examine the effectiveness of CHAMPS II, an inclusive, choice-based physical activity promotion program to increase lifetime physical activity levels of seniors. CHAMPS guided participants to choose activities that took into account their health, preferences, and abilities. It offered information on ways for them to exercise safely, motivate themselves, overcome barriers, and develop a balanced exercise regimen. METHODS: A 1-year randomized controlled trial was conducted with physically underactive seniors in a multispecialty group practice. Changes in self-reported physical activity by group were evaluated using ANCOVA, controlling for age and sex. RESULTS: Of 173 randomized subjects, 164 (95%) completed the trial. Subjects were aged 65 to 90 years (M = 74, SD = 6); 66% were female. The intervention group increased estimated caloric expenditure by 487 calories/week in moderate (or greater) intensity activities (MET >/= 3.0; p <.001) and by 687 calories/week in physical activities of any intensity (p <.001). Control group changes were negligible. Between-group analyses found that the changes were significantly different in both measures (p values <.05). Overweight persons especially benefited from this program. The program was as effective for women, older adults (75+), and those who did not set aside time to exercise at baseline. CONCLUSIONS: The program led to meaningful physical activity increases. Individually tailored programs to encourage lifestyle changes in seniors may be effective and applicable to health care and community settings. PMID- 11487599 TI - Evidence for a shift in the Th-1 to Th-2 cytokine response associated with chronic stress and aging. AB - BACKGROUND: A number of studies have shown that the chronic stress of caring for persons with dementia can have significant immunological consequences as demonstrated by the down-regulation/dysregulation of the cellular immune response. METHODS: Utilizing flow cytometry to measure the percentages and absolute numbers of CD-4(+) and CD-8(+) T lymphocytes producing the cytokines indicative of Th-1, Tc1 and Th-2, and Tc2 cells, we compared spousal caregivers and control subjects. The expression of interleukin-2 (IL-2), interferon gamma (IFN-gamma), and interleukin-10 (IL-10) in the cytoplasm of CD-4(+) and CD-8(+) lymphocytes was assessed. RESULTS: Neither stress nor age was significantly related to the percentage or number of IFNgamma(+)/CD-8(+), IL-2(+)/CD-8(+) cells, or IFNgamma(+), IL-2(+), CD-4(+) cells. However, the percentage of IL 10(+) cells was higher in lymphocytes obtained from caregivers than control subjects. In addition, the significant interaction between stress and aging for IL-10(+)/CD-4(+) and IL-10(+)/CD-8(+) cells demonstrated that the difference between caregivers and control subjects was age dependent; the difference between caregivers and control subjects was substantially larger in younger individuals than in older individuals. CONCLUSIONS: The data are consistent with previous reports on acute stress and suggest that there may also be a shift from a Th-1 to a Th-2 response associated with a chronic stressor such as caregiving. This shift could have implications for an individual's responses to pathogens. PMID- 11487600 TI - Anthropometric changes over 5 years in elderly Canadians by age, gender, and cognitive status. AB - BACKGROUND: Numerous changes in body composition occur with aging. This study reports on secondary analyses of data from a subsample of institutionalized and free-living elderly Canadians taking part in both phases of the Canadian Study of Health and Aging (CSHA-1 and CHSA-2; n = 10,263) to document and examine correlates of the evolution of anthropometric characteristics over a 5-year period. METHODS: In CSHA-1, community-dwelling (n = 1464) and institutionalized (n = 963) participants' height and weight were measured in clinics. Surviving participants were remeasured in CSHA-2; valid data were available for 487 community-dwelling respondents (66.9% of those seen in clinics in CSHA-2) and 140 institutionalized participants (46.9% of those reassessed). Body mass index (BMI = weight [kg]/height [m(2)]) was calculated. Paired t tests were used to test changes over the interval, and repeated-measures multivariate analysis of variance was used to examine the extent of differences within and across categories. RESULTS: The average weight loss between study phases in community dwelling and institutionalized participants was approximately 2 kg (p <.001). In institutions, this was statistically significant in most stratification categories, as was the case in community-dwelling participants (by gender, age, dementia screening score, and cognitive diagnosis). Among those who were cognitively intact, the greatest weight losses occurred in participants under 90 years old and in those aged 70 to 79 years with a diagnosis of dementia (p <.01). Stature decreased more in institutionalized (2 cm) than in community-dwelling participants (1.4 cm). In institutions, this was significant among the oldest men (p <.005), while in the community there were no differences in the extent of height lost in all stratification categories. The average BMI was largely stable. CONCLUSIONS: Body weight and stature declined with aging among elderly Canadian CSHA participants, particularly in the very old and those with dementia. Such longitudinal anthropometric data are needed along with information on dietary intakes, and medical, cognitive, and functional measures to plan interventions geared to maximizing nutritional and overall health in the elderly population, whatever their cognitive status. PMID- 11487601 TI - The interacting effects of cognitive demand and recovery of postural stability in balance-impaired elderly persons. AB - BACKGROUND: Although postural recovery is attentionally demanding in healthy elderly persons, an inability to recover balance due to competition for attentional resources between the postural system and a second task could contribute to falls in older adults with poor balance. This study examined the attentional demands of balance recovery from a mild postural disturbance in balance-impaired elderly persons. A second purpose of this research was to determine the effect of performing a cognitive task on the recovery of balance in balance-impaired elderly persons. METHODS: Fifteen healthy older adults and 13 older adults with clinical balance impairment were exposed to balance disturbances by means of sudden movement of a platform on which they stood. A dual-task paradigm where postural recovery served as the primary task and verbal reaction time to auditory tones served as the secondary task was used to assess attentional demand. To determine the effect of the cognitive task on postural recovery, kinetic, kinematic, and neuromuscular measures of a feet-in-place response were investigated. RESULTS: Balance recovery using a feet-in-place response was attentionally demanding in both groups of older adults and was more demanding in balance-impaired than in healthy elderly persons. With the concurrent performance of a cognitive task, balance-impaired elderly persons took longer to stabilize their center of pressure and regain balance than in a single task, while healthy elderly persons showed no change between conditions. In addition, only balance-impaired elderly individuals had a greater center-of pressure resultant velocity during recovery in a dual-task compared with a single task situation. CONCLUSIONS: The ability to recover balance using a feet-in-place response was more attentionally demanding in balance-impaired than in healthy elderly persons. The recovery of balance was also slower and less efficient in balance-impaired elderly persons when simultaneously performing a cognitive task, whereas the ability of healthy elderly individuals to recover was not influenced by concurrent task demands. This suggests that dual-task performance may contribute to postural instability and falls in balance-impaired elderly individuals. PMID- 11487602 TI - The efficacy of exercise as a long-term antidepressant in elderly subjects: a randomized, controlled trial. AB - BACKGROUND: Pharmacological treatment of depression in geriatric patients is often difficult. Although unsupervised exercise has been shown to benefit younger depressed patients, there is no evidence that unsupervised exercise can be used as a maintenance treatment for depression in elderly patients. Our aim was to test the feasibility and efficacy of unsupervised exercise as a long-term treatment for clinical depression in elderly patients. METHODS: We studied 32 subjects (71.3 +/- 1.2 years of age, mean +/- SE) in a 20-week, randomized, controlled trial, with follow-up at 26 months. Subjects were community-dwelling patients with major or minor depression or dysthymia. Exercisers engaged in 10 weeks of supervised weight-lifting exercise followed by 10 weeks of unsupervised exercise. Controls attended lectures for 10 weeks. No contact was made with either group after 20 weeks until final follow-up. Blinded assessment was made with the Beck Depression Inventory (BDI), the Philadelphia Geriatric Morale Scale, and Ewart's Self Efficacy Scale at 20 weeks and with the BDI and physical activity questionnaire at 26 months. RESULTS: Patients randomized to the exercise condition completed 18 +/- 2 sessions of unsupervised exercise during Weeks 10 to 20. The BDI was significantly reduced at both 20 weeks and 26 months of follow-up in exercisers compared with controls (p <.05-.001). At the 26-month follow-up, 33% of the exercisers were still regularly weight lifting, versus 0% of controls (p <.05). CONCLUSIONS: Unsupervised weight-lifting exercise maintains its antidepressant effectiveness at 20 weeks in depressed elderly patients. Long-term changes in exercise behavior are possible in some patients even without supervision. PMID- 11487603 TI - The association of depression and mortality in elderly persons: a case for multiple, independent pathways. AB - BACKGROUND: The evidence for an association between depression and mortality among community-dwelling elderly persons remains inconclusive, although it is well established for younger individuals. Extant studies suggest that this association weakens when adjusted for potential confounding factors, especially functional impairment. A cohort of elderly subjects followed for 3 years was analyzed to determine the association of depression and 3-year mortality, controlling for the major known risk factors for mortality in the elderly population. METHODS: Information on depression (CES-D scores), mortality, demographics, body mass index, chronic disease, smoking history, cognitive impairment, functional impairment, self-rated health, and social support was obtained from a stratified probability-based sample of community-dwelling elderly persons, with equal distribution between African Americans and whites in the Piedmont of North Carolina. Descriptive statistics were calculated, and logistic regression was used for a series of models with progressively more control variables. RESULTS: The unadjusted relative odds of mortality among depressed subjects at baseline was 1.98 over 3 years of follow-up. Inclusion of age, gender, and race into the model did not reduce the relative odds. When chronic disease and health habits, cognitive impairment, functional impairment, and social support were added to the model, the odds ratios for mortality with depression were 1.74, 1.69, 1.29, and 1.21, respectively. This decrease in odds ratios was not observed for other variables in the model when additional variables were added. CONCLUSIONS: The estimated odds of dying if depressed moved toward unity as other risk factors for mortality were controlled. Unlike other known risk factors for mortality in the elderly population, depression appears to be associated with mortality through a number of independent mechanisms, perhaps through complex feedback loops. PMID- 11487604 TI - Nonsteroidal anti-inflammatory therapy after eccentric exercise in healthy older individuals. AB - BACKGROUND: Aging is associated with greater susceptibility to muscle injury and soreness after exercise. Although elderly persons regularly consume nonsteroidal anti-inflammatory drugs (NSAIDs), it is not clear that NSAIDs alleviate muscle dysfunction and/or inflammation following injurious exercise. METHODS: In this double-blind crossover study, 10 men and 5 women (aged 60 +/- 2 years, mean +/- SE) consumed naproxen sodium or placebo for 10 days after performing 64 unilateral eccentric (ECC) knee extensions using 75% of the ECC 1-repetition maximum. Strength was measured before, 3 days after, and 10 days after each bout. Injury and soreness were assessed using magnetic resonance images of m. quadriceps femoris (QF) and a visual analog scale. RESULTS: Three days after exercise, concentric strength loss was greater for placebo (-32 +/- 9%) than NSAID (-6 +/- 8%; p =.0064). Likewise, isometric strength declined less for NSAID than placebo (-12 +/- 7% vs -24 +/- 4%; p =.0213), and thigh soreness while rising from a chair was greater for placebo (p < or =.0393) than NSAID (43 +/- 7 mm vs 26 +/- 7 mm). QF cross-sectional area (cm(2)) showing elevated T(2) was 27% and 35% greater (p < or =.0096) for placebo on Days 3 and 10, respectively. CONCLUSIONS: Naproxen sodium attenuated muscle injury, strength loss, and soreness following ECC exercise in older individuals and may be beneficial during the early stages of increased physical activity. PMID- 11487605 TI - Survey of primary care physicians' approach to gastroesophageal reflux disease in elderly patients. AB - BACKGROUND: Although gastroesophageal reflux disease (GERD) is a common condition, little is known regarding physicians' approach to the diagnosis and management of GERD in elderly patients. METHODS: We surveyed by facsimile a random sample of 14,000 practicing primary care physicians throughout the United States. Physicians were questioned using a case-based format about the approach to a symptomatic patient with GERD including the use of empiric therapy, the role of diagnostic testing, and the drugs of choice to treat GERD. RESULTS: A total of 2241 surveys (16%) was returned and tabulated. Most respondents were either internists (37%) or family practice physicians (56%) in solo or group practice, and 74% had been in practice for 11 or more years. There were 1980 (90%) respondents who evaluated more than 6 patients per week with GERD. Empiric therapy was commonly recommended for the symptomatic patient, most often in a step-up approach beginning with H(2)-receptor blockers. Diagnostic testing, usually endoscopy, was recommended appropriately in patients with alarm symptoms. Proton-pump inhibitors were most often recommended for patients failing to respond to over-the-counter H(2)-receptor blockers and for those with endoscopic esophagitis; the use of cisapride in combination with H(2)-receptor blockers was also commonly recommended in these scenarios. CONCLUSIONS: The management of symptomatic GERD in elderly patients appears similar to the management of GERD in other patients. Empiric therapy was frequently recommended in a step-up approach, and diagnostic testing was appropriate. Combination therapy with cisapride and an acid-reducing agent was commonly recommended. PMID- 11487606 TI - Skilled finger movement exercise improves hand function. AB - BACKGROUND: Aging is accompanied by a marked decline in muscle strength and ability to maintain steady submaximal force. Studies have shown that exercise programs can improve age-related regression of hand function in elderly individuals. The purpose of this study was to train elderly subjects to perform skilled finger movements and to evaluate the changes in hand function involving skillful use of finger pinch. METHODS: Grip strength, maximum pinch force (MPF), steadiness of pinch force at 5%, 10%, and 20% MPF, M wave, and Hoffman (H) reflex were measured. Fourteen elderly subjects were trained with skilled finger movements, and their performance involving finger pinch was measured. RESULTS: Compared with untrained elderly subjects, the trained older adults significantly (p <.05) improved their ability to control submaximal pinch force, to maintain a steady hand posture, and to relocate a small object quickly with finger grip. The amplitude of H reflex increased significantly for the trained group. CONCLUSIONS: Skilled finger movement training improves the ability to control submaximal pinch force, hand steadiness, and manual speed in elderly subjects; these improvements may be due to training-induced adaptations in the central and peripheral nervous systems. PMID- 11487607 TI - The acute antihyperalgesic action of nonsteroidal, anti-inflammatory drugs and release of spinal prostaglandin E2 is mediated by the inhibition of constitutive spinal cyclooxygenase-2 (COX-2) but not COX-1. AB - Western blots show the constitutive expression of COX-1 and COX-2 in the rat spinal dorsal and ventral horns and in the dorsal root ganglia. Using selective inhibitors of cyclooxygenase (COX) isozymes, we show that in rats with chronic indwelling intrathecal catheters the acute thermal hyperalgesia evoked by the spinal delivery of substance P (SP; 20 nmol) or NMDA (2 nmol) and the thermal hyperalgesia induced by the injection of carrageenan into the paw are suppressed by intrathecal and systemic COX-2 inhibitors. The intrathecal effects are dose dependent and stereospecific. In contrast, a COX-1 inhibitor given systemically, but not spinally, reduced carrageenan-evoked thermal hyperalgesia but had no effect by any route with spinal SP hyperalgesia. Using intrathecal loop dialysis catheters, we showed that intrathecal SP would enhance the release of prostaglandin E(2) (PGE(2)). This intrathecally evoked release of spinal PGE(2) was diminished by systemic delivery of nonspecific COX and COX-2-selective inhibitors, but not a COX-1-selective inhibitor. Given at systemic doses that block SP- and carrageenan-evoked hyperalgesia, COX-2, but not COX-1, inhibitors reduced spinal SP-evoked PGE(2) release. Thus, constitutive spinal COX-2, but not COX-1, is an important contributor to the acute antihyperalgesic effects of spinal as well as systemic COX-2 inhibitors. PMID- 11487608 TI - A prosurvival function for the p75 receptor death domain mediated via the caspase recruitment domain receptor-interacting protein 2. AB - In addition to promoting cell survival, neurotrophins also can elicit apoptosis in restricted cell types. Recent results indicate that nerve growth factor (NGF) can induce Schwann cell death via engagement of the p75 neurotrophin receptor. Here we describe a novel interaction between the p75 receptor and receptor interacting protein 2, RIP2 (RICK/CARDIAK), that accounts for the ability of neurotrophins to choose between a survival-versus-death pathway. RIP2, an adaptor protein with a serine threonine kinase and a caspase recruitment domain (CARD), is highly expressed in dissociated Schwann cells and displays an endogenous association with p75. RIP2 binds to the death domain of p75 via its CARD domain in an NGF-dependent manner. The introduction of RIP2 into Schwann cells deficient in RIP2 conferred NGF-dependent nuclear transcription factor-kappaB (NF-kappaB) activity and decreased the cell death induced by NGF. Conversely, the expression of a dominant-negative version of RIP2 protein resulted in a loss of NGF-induced NF-kappaB induction and increased NGF-mediated cell death. These results indicate that adaptor proteins like RIP2 can provide a bifunctional switch for cell survival or cell death decisions mediated by the p75 neurotrophin receptor. PMID- 11487609 TI - Altered processing of pro-orphanin FQ/nociceptin and pro-opiomelanocortin-derived peptides in the brains of mice expressing defective prohormone convertase 2. AB - The bioactivity of neuropeptides can be regulated by a variety of post translational modifications, including proteolytic processing. Here, gene targeted mice producing defective prohormone convertase 2 (PC2) were used to examine the post-translational processing of two neuroendocrine prohormones, pro opiomelanocortin (POMC) and pro-orphanin FQ (pOFQ)/nociceptin (N), in the brain. Reversed-phase HPLC and gel-exclusion chromatography were combined with specific radioimmunoassays to analyze the processing patterns of these two prohormones in the hypothalamus and the amygdala. In the case of POMC, the lack of PC2 activity completely prevented carboxy-shortening of beta-endorphins and greatly diminished conversion of beta-lipotropin to gamma-lipotropin and beta-endorphin. Although conversion of beta-lipotropin to beta-endorphin decreased, the lack of PC2 activity caused an increase in beta-lipotropin and beta-endorphin levels in the mutant animals, but no increases in POMC or biosynthetic intermediates were seen. The extent of OFQ/N production was significantly lower in PC2-deficient mice and there was an accumulation of relatively large amounts of pOFQ/N and biosynthetic intermediates. These results demonstrate that PC2 is directly involved in the biogenesis of two brain neuropeptides in vivo and suggest that the specific prohormone and cellular context influences neuropeptide processing by PCs. PMID- 11487610 TI - Mutations in the Caenorhabditis elegans serotonin reuptake transporter MOD-5 reveal serotonin-dependent and -independent activities of fluoxetine. AB - We isolated two mutants defective in the uptake of exogenous serotonin (5-HT) into the neurosecretory motor neurons of Caenorhabditis elegans. These mutants were hypersensitive to exogenous 5-HT and hyper-responsive in the experience dependent enhanced slowing response to food modulated by 5-HT. The two allelic mutations defined the gene mod-5 (modulation of locomotion defective), which encodes the only serotonin reuptake transporter (SERT) in C. elegans. The selective serotonin reuptake inhibitor fluoxetine (Prozac) potentiated the enhanced slowing response, and this potentiation required mod-5 function, establishing a 5-HT- and SERT-dependent behavioral effect of fluoxetine in C. elegans. By contrast, other responses of C. elegans to fluoxetine were independent of MOD-5 SERT and 5-HT. Further analysis of the MOD-5-independent behavioral effects of fluoxetine could lead to the identification of novel targets of fluoxetine and could facilitate the development of more specific human pharmaceuticals. PMID- 11487611 TI - On the contribution of the first transmembrane domain to whole-cell current through an ATP-gated ionotropic P2X receptor. AB - Scanning cysteine mutagenesis was used to identify potential pore-forming residues in and around the first transmembrane domains of ionotropic P2X(2) receptor subunits. Twenty-eight unique cysteine-substituted mutants (R28C-Y55C) were individually expressed in HEK293 cells by lipofection. Twenty-three of these were functional as assayed by application of ATP to transfected voltage-clamped cells. Individual mutants varied in their sensitivity to ATP; otherwise, currents through functional mutant receptors resembled those of the homomeric wild-type (WT) receptor. In five (H33C, R34C, I50C, K53C, and S54C) of 23 functional mutants, coapplication of 30 microm ATP and 500 nm Ag(+) irreversibly inhibited inward current evoked by subsequent applications of ATP alone. These inhibitions did not result in a lateral shift in the agonist concentration-response curve and are unlikely to involve a modification of the agonist binding site. Two (K53C and S54C) of the five residues modified by Ag(+) applied in the presence of ATP when the channels were gating were also modified by 1 mm (2 aminoethyl)methanethiosulfonate applied in the absence of ATP when the channels were closed. These data suggest that domains near either end of the first transmembrane domain influence ion conduction through the pore of the P2X(2) receptor. PMID- 11487612 TI - A dominant negative inhibitor of the Egr family of transcription regulatory factors suppresses cerebellar granule cell apoptosis by blocking c-Jun activation. AB - To investigate the role of the Egr family of transcription regulatory factors in neuronal apoptosis, we examined the effect of a dominant negative Egr inhibitor construct in a well characterized in vitro paradigm, cerebellar granule cell death induced by withdrawal of depolarizing concentrations of extracellular potassium. We found that this apoptotic stimulus increases the activity of a reporter gene driven by the Egr response element and that a dominant negative inhibitor of Egr-mediated transcription blocks granule cell apoptosis. In contrast, apoptosis of immature granule cells induced by cytosine arabinoside is not inhibited by the Egr inhibitor construct. Because activation of c-Jun is an essential step in granule cell death induced by potassium deprivation, but not cytosine arabinoside, we asked whether the Egr inhibitor acts by influencing c Jun activation or its ability to induce apoptosis. We found that the Egr inhibitor does not block the ability of a constitutively active c-Jun construct to induce apoptosis in these cells but does suppress activation of c-Jun-mediated transcription induced by lowering extracellular potassium concentration. Furthermore, the Egr inhibitor blocks the ability of MEKK1 [mitogen-activated protein kinase (MAPK) kinase kinase 1], an upstream kinase capable of stimulating the JNK (c-Jun N-terminal protein kinase)-c-Jun pathway, to induce apoptosis and activate c-Jun. Together, these studies indicate that the Egr family of transcription factors plays a critical role in neuronal apoptosis and identify c Jun activation as an important downstream target of the Egr family in this process. PMID- 11487613 TI - Paradoxical role of large-conductance calcium-activated K+ (BK) channels in controlling action potential-driven Ca2+ entry in anterior pituitary cells. AB - Activation of high-conductance Ca(2+)-activated K(+) (BK) channels normally limits action potential duration and the associated voltage-gated Ca(2+) entry by facilitating membrane repolarization. Here we report that BK channel activation in rat pituitary somatotrophs prolongs membrane depolarization, leading to the generation of plateau-bursting activity and facilitated Ca(2+) entry. Such a paradoxical role of BK channels is determined by their rapid activation by domain Ca(2+), which truncates the action potential amplitude and thereby limits the participation of delayed rectifying K(+) channels during membrane repolarization. Conversely, pituitary gonadotrophs express relatively few BK channels and fire single spikes with a low capacity to promote Ca(2+) entry, whereas an elevation in BK current expression in a gonadotroph model system leads to the generation of plateau-bursting activity and high-amplitude Ca(2+) transients. PMID- 11487614 TI - Amphetamine distorts stimulation-dependent dopamine overflow: effects on D2 autoreceptors, transporters, and synaptic vesicle stores. AB - Amphetamine (AMPH) is known to raise extracellular dopamine (DA) levels by inducing stimulation-independent DA efflux via reverse transport through the DA transporter and by inhibiting DA re-uptake. In contrast, recent studies indicate that AMPH decreases stimulation-dependent vesicular DA release. One candidate mechanism for this effect is the AMPH-mediated redistribution of DA from vesicles to the cytosol. In addition, the inhibition of stimulation-dependent release may occur because of D2 autoreceptor activation by DA that is released via reverse transport. We used the D2 receptor antagonist sulpiride and mice lacking the D2 receptor to address this issue. To evaluate carefully AMPH effects on release and uptake, we recorded stimulated DA overflow in striatal slices by using continuous amperometry and cyclic voltammetry. Recordings were fit by a random walk simulation of DA diffusion, including uptake with Michaelis-Menten kinetics, that provided estimates of DA concentration and uptake parameters. AMPH (10 microm) promoted the overflow of synaptically released DA by decreasing the apparent affinity for DA uptake (K(m) increase from 0.8 to 32 microm). The amount of DA released per pulse, however, was decreased by 82%. This release inhibition was prevented partly by superfusion with sulpiride (47% inhibition) and was reduced in D2 mutant mice (23% inhibition). When D2 autoreceptor activation was minimal, the combined effects of AMPH on DA release and uptake resulted in an enhanced overflow of exocytically released DA. Such enhancement of stimulation-dependent DA overflow may occur under conditions of low D2 receptor activity or expression, for example as a result of AMPH sensitization. PMID- 11487615 TI - Metabotropic glutamate receptors 1 and 5 differentially regulate CA1 pyramidal cell function. AB - The activation of group I metabotropic glutamate receptors (mGluRs) produces a variety of actions that lead to alterations in excitability and synaptic transmission in the CA1 region of the hippocampus. The group I mGluRs, mGluR1 and mGluR5, are activated selectively by (S)-3,5-dihydroxyphenylglycine (DHPG). To identify which of these mGluR subtypes are responsible for the various actions of DHPG in area CA1, we took advantage of two novel subtype-selective antagonists. (S)-(+)-alpha-amino-a-methylbenzeneacetic acid (LY367385) is a potent competitive antagonist that is selective for mGluR1, whereas 2-methyl-6-(phenylethynyl) pyridine (MPEP) is a potent noncompetitive antagonist that is selective for mGluR5. The use of these compounds in experiments with whole-cell patch-clamp recording and Ca(2+)-imaging techniques revealed that each group I mGluR subtype plays distinct roles in regulating the function of CA1 pyramidal neurons. The block of mGluR1 by LY367385 suppressed the DHPG-induced increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) and the direct depolarization of CA1 hippocampal neurons. In addition, the increase in the frequency of spontaneous IPSCs (sIPSCs) caused by the DHPG-induced depolarization of inhibitory interneurons also was blocked by LY367385, as was the DHPG-induced inhibition of transmission at the Schaffer collateral-->CA1 synapse. On the other hand, the block of mGluR5 by MPEP antagonized the DHPG-induced suppression of the Ca(2+) activated potassium current (I(AHP)) and potentiation of the NMDA receptor. Finally, antagonism of the DHPG-induced suppression of evoked IPSCs required the blockade of both mGluR1 and mGluR5. These data suggest that mGluR1 and mGluR5 play distinct roles in the regulation of the excitability of hippocampal CA1 pyramidal neurons. PMID- 11487616 TI - GABA enhances transmission at an excitatory glutamatergic synapse. AB - GABA mediates both presynaptic and postsynaptic inhibition at many synapses. In contrast, we show that GABA enhances transmission at excitatory synapses between the lateral gastric and medial gastric motor neurons and the gastric mill 6a and 9 (gm6a, gm9) muscles and between the lateral pyloric motor neuron and pyloric 1 (p1) muscles in the stomach of the lobster Homarus americanus. Two-electrode current-clamp or voltage-clamp techniques were used to record from muscle fibers. The innervating nerves were stimulated to evoke excitatory junctional potentials (EJPs) or excitatory junctional currents. Bath application of GABA first decreased the amplitude of evoked EJPs in gm6a and gm9 muscles, but not the p1 muscle, by activating a postjunctional conductance increase that was blocked by picrotoxin. After longer GABA applications (5-15 min), the amplitudes of evoked EJPs increased in all three muscles. This increase persisted in the presence of picrotoxin. beta-(Aminomethyl)-4-chlorobenzenepropanoic acid (baclofen) was an effective agonist for the GABA-evoked enhancement but did not increase the postjunctional conductance. Muscimol activated a rapid postsynaptic conductance but did not enhance the amplitude of the nerve-evoked EJPs. GABA had no effect on iontophoretic responses to glutamate and decreased the coefficient of variation of nerve-evoked EJPs. In the presence or absence of tetrodotoxin, GABA increased the frequency but not the amplitude of miniature endplate potentials. These data suggest that GABA acts presynaptically via a GABA(B)-like receptor to increase the release of neurotransmitter. PMID- 11487617 TI - Neuronal Ca(V)1.3alpha(1) L-type channels activate at relatively hyperpolarized membrane potentials and are incompletely inhibited by dihydropyridines. AB - L-type calcium channels regulate a diverse array of cellular functions within excitable cells. Of the four molecularly defined subclasses of L-type Ca channels, two are expressed ubiquitously in the mammalian nervous system (Ca(V)1.2alpha(1) and Ca(V)1.3alpha(1)). Despite diversity at the molecular level, neuronal L-type channels are generally assumed to be functionally and pharmacologically similar, i.e., high-voltage activated and highly sensitive to dihydropyridines. We now show that Ca(V)1.3alpha(1) L-type channels activate at membrane potentials approximately 25 mV more hyperpolarized, compared with Ca(V)1.2alpha(1). This unusually negative activation threshold for Ca(V)1.3alpha(1) channels is independent of the specific auxiliary subunits coexpressed, of alternative splicing in domains I-II, IVS3-IVS4, and the C terminus, and of the expression system. The use of high concentrations of extracellular divalent cations has possibly obscured the unique voltage-dependent properties of Ca(V)1.3alpha(1) in certain previous studies. We also demonstrate that Ca(V)1.3alpha(1) channels are pharmacologically distinct from Ca(V)1.2alpha(1). The IC(50) for nimodipine block of Ca(V)1.3alpha(1) L-type calcium channel currents is 2.7 +/- 0.3 microm, a value 20-fold higher than the concentration required to block Ca(V)1.2alpha(1). The relatively low sensitivity of the Ca(V)1.3alpha(1) subunit to inhibition by dihydropyridine is unaffected by alternative splicing in the IVS3-IVS4 linker. Our results suggest that functional and pharmacological criteria used commonly to distinguish among different Ca currents greatly underestimate the biological importance of L-type channels in cells expressing Ca(v)1.3alpha(1). PMID- 11487618 TI - Nav1.3 sodium channels: rapid repriming and slow closed-state inactivation display quantitative differences after expression in a mammalian cell line and in spinal sensory neurons. AB - Although rat brain Nav1.3 voltage-gated sodium channels have been expressed and studied in Xenopus oocytes, these channels have not been studied after their expression in mammalian cells. We characterized the properties of the rat brain Nav1.3 sodium channels expressed in human embryonic kidney (HEK) 293 cells. Nav1.3 channels generated fast-activating and fast-inactivating currents. Recovery from inactivation was relatively rapid at negative potentials (<-80 mV) but was slow at more positive potentials. Development of closed-state inactivation was slow, and, as predicted on this basis, Nav1.3 channels generated large ramp currents in response to slow depolarizations. Coexpression of beta3 subunits had small but significant effects on the kinetic and voltage-dependent properties of Nav1.3 currents in HEK 293 cells, but coexpression of beta1 and beta2 subunits had little or no effect on Nav1.3 properties. Nav1.3 channels, mutated to be tetrodotoxin-resistant (TTX-R), were expressed in SNS-null dorsal root ganglion (DRG) neurons via biolistics and were compared with the same construct expressed in HEK 293 cells. The voltage dependence of steady-state inactivation was approximately 7 mV more depolarized in SNS-null DRG neurons, demonstrating the importance of background cell type in determining physiological properties. Moreover, consistent with the idea that cellular factors can modulate the properties of Nav1.3, the repriming kinetics were twofold faster in the neurons than in the HEK 293 cells. The rapid repriming of Nav1.3 suggests that it contributes to the acceleration of repriming of TTX-sensitive (TTX-S) sodium currents that are seen after peripheral axotomy of DRG neurons. The relatively rapid recovery from inactivation and the slow closed-state inactivation kinetics of Nav1.3 channels suggest that neurons expressing Nav1.3 may exhibit a reduced threshold and/or a relatively high frequency of firing. PMID- 11487619 TI - Contribution of the Na-K-Cl cotransporter on GABA(A) receptor-mediated presynaptic depolarization in excitatory nerve terminals. AB - GABA(A) receptor-mediated responses manifest as either hyperpolarization or depolarization according to the intracellular Cl(-) concentration ([Cl(-)](i)). Here, we report a novel functional interaction between the Na-K-Cl cotransporter (NKCC) and GABA(A) receptor actions on glutamatergic presynaptic nerve terminals projecting to ventromedial hypothalamic (VMH) neurons. The activation of presynaptic GABA(A) receptors depolarizes the presynaptic nerve terminals and facilitates spontaneous glutamate release by activating TTX-sensitive Na(+) channels and high-threshold Ca(2+) channels. This depolarizing action of GABA was caused by an outwardly directed Cl(-) driving force for GABA(A) receptors; that is, the [Cl(-)](i) of glutamatergic nerve terminals was higher than that predicted for a passive distribution. The higher [Cl(-)](i) was generated by bumetanide-sensitive NKCCs and was responsible for the GABA-induced presynaptic depolarization. Thus, GABA(A) receptor-mediated modulation of spontaneous glutamatergic transmission may contribute to the development and regulation of VMH function as well as to the excitability of VMH neurons themselves. PMID- 11487620 TI - Experimental localization of Kv1 family voltage-gated K+ channel alpha and beta subunits in rat hippocampal formation. AB - In the mammalian hippocampal formation, dendrotoxin-sensitive voltage-gated K(+) (Kv) channels modulate action potential propagation and neurotransmitter release. To explore the neuroanatomical basis for this modulation, we used in situ hybridization, coimmunoprecipitation, and immunohistochemistry to determine the subcellular localization of the Kv channel subunits Kv1.1, Kv1.2, Kv1.4, and Kvbeta2 within the adult rat hippocampus. Although mRNAs encoding all four of these Kv channel subunits are expressed in the cells of origin of each major hippocampal afferent and intrinsic pathway, immunohistochemical staining suggests that the encoded subunits are associated with the axons and terminal fields of these cells. Using an excitotoxin lesion strategy, we explored the subcellular localization of these subunits in detail. We found that ibotenic acid lesions of the entorhinal cortex eliminated Kv1.1 and Kv1.4 immunoreactivity and dramatically reduced Kv1.2 and Kvbeta2 immunoreactivity in the middle third of the dentate molecular layer, indicating that these subunits are located on axons and terminals of entorhinal afferents. Similarly, ibotenic acid lesions of the dentate gyrus eliminated Kv1.1 and Kv1.4 immunoreactivity in the stratum lucidum of CA3, indicating that these subunits are located on mossy fiber axons. Kainic acid lesions of CA3 dramatically reduced Kv1.1 immunoreactivity in the stratum radiatum of CA1-CA3, indicating that Kv1.1 immunoreactivity in these subfields is associated with the axons and terminals of the Schaffer collaterals. Together with the results of coimmunoprecipitation analyses, these data suggest that action potential propagation and glutamate release at excitatory hippocampal synapses are directly modulated by Kv1 channel complexes predominantly localized on axons and nerve terminals. PMID- 11487621 TI - Gating and braking of short- and long-term modulatory effects by interactions between colocalized neuromodulators. AB - Spinal locomotor networks in the lamprey are modulated by tachykinin neuropeptides. A single 10 min application of the tachykinin substance P evokes a short-term ( approximately 1 hr) presynaptic facilitation of glutamate release and the postsynaptic potentiation of NMDA responses. The latter effect induces a long-term (>24 hr) protein synthesis-dependent increase in the frequency of network activity. Tachykinins are contained in a ventromedial spinal plexus into which the medial dendrites of network neurons project. Neurons in this plexus also contain colocalized dopamine and 5-HT. Here, dynamic plasticity evoked by modulator interactions has been examined by investigating the effects of 5-HT and dopamine on specific cellular, synaptic, and network effects of substance P. Preapplied 5-HT blocked the substance P-mediated increase in the network burst frequency and the potentiation of NMDA-evoked cellular responses that underlies its induction. 5-HT also blocked the presynaptic facilitation of glutamatergic synaptic transmission by substance P. The presynaptic, but not postsynaptic, effect of 5-HT was reduced by the protein phosphatase 2B inhibitor cypermethrin. Dopamine did not directly modulate the effects of substance P. However, it reduced the presynaptic interactive effect of 5-HT and thus gated the presynaptic potentiation of glutamatergic inputs by substance P. However, the substance P mediated potentiation of NMDA responses was not gated by dopamine, and thus the long-term network modulation was not induced. Neuromodulator effects and their interactions can thus be modulated. By selecting components from the modulatory repertoire of substance P, these interactions evoke dynamic changes in short- and long-term synaptic and network plasticity. PMID- 11487622 TI - Interactions between apolipoprotein E gene and dietary alpha-tocopherol influence cerebral oxidative damage in aged mice. AB - Cerebral oxidative damage is a feature of aging and is increased in a number of neurodegenerative diseases. We pursued the gene-environment interaction of lack of apolipoprotein E (apoE) and modulation of dietary alpha-tocopherol on cerebral oxidative damage in aged male and female mice by quantifying the major isomers of cerebral isoprostanes, derived from arachidonic acid (AA) oxidation, and neuroprostanes, derived from docosahexaenoic acid (DHA) oxidation. Mice fed alpha tocopherol-deficient, normal, or -supplemented diet had undetectable, 4486 +/- 215, or 6406 +/- 254 ng of alpha-tocopherol per gram of brain tissue (p < 0.0001), respectively. Two factors, male gender and lack of apoE, combined to increase cerebral AA oxidation by 28%, whereas three factors, male gender, lack of apoE, and deficiency in alpha-tocopherol, combined to increase cerebral DHA oxidation by 81%. alpha-Tocopherol supplementation decreased cerebral isoprostanes but not neuroprostanes and enhanced DHA, but not AA, endoperoxide reduction in vivo and in vitro. These results demonstrated that the interaction of gender, inherited susceptibilities, and dietary alpha-tocopherol contributed differently to oxidative damage to cerebral AA and DHA in aged mice. PMID- 11487623 TI - A high-efficiency protein transduction system demonstrating the role of PKA in long-lasting long-term potentiation. AB - Proteins and peptides have been demonstrated to penetrate across the plasma membrane of eukaryotic cells by protein transduction domains. We show that protein transduction by 11 arginine (11R) is an efficient method of delivering proteins into the neurons of brain slices. Here, we demonstrate that PKA inhibitory peptide, fused with 11R and nuclear localization signal, delivers the peptide exclusively into the nuclear compartment of neurons in brain slices. This inhibitory peptide blocked both cAMP responsive element-binding protein phosphorylation and long-lasting long-term potentiation (LTP) induction, but not early LTP. These results highlight transduction of proteins and peptides into specific neuronal subcellular compartments in brain slices as a powerful tool for studying neuronal plasticity. PMID- 11487624 TI - Activation of silent synapses by rapid activity-dependent synaptic recruitment of AMPA receptors. AB - Many recent studies have shown that excitatory synapses can contain NMDA receptor responses in the absence of functional AMPA receptors and are therefore postsynaptically silent at resting membrane potentials. The activation of silent synapses via the rapid acquisition of AMPA receptor responses may be important in synaptic plasticity and neuronal development. Our recent immunocytochemical studies that used cultured hippocampal neurons have provided evidence for "morphological silent synapses" that physically contain NMDA receptors but no AMPA receptors. Here we show that the activation of NMDA receptors by spontaneous synaptic activity results in the rapid recruitment of AMPA receptors into these morphological silent synapses within minutes. In parallel, we find a significant increase in the frequency of AMPA receptor-mediated miniature EPSCs (mEPSCs). NMDA receptor activation also results in a mobilization of calcium/calmodulin (CaM) kinase II to synapses and an increase in the phosphorylation of surface AMPA receptors on the major CaM kinase II phosphorylation site. These results demonstrate that AMPA receptors can be modified and recruited rapidly to silent synapses via the activation of NMDA receptors by spontaneous synaptic activity. PMID- 11487625 TI - Molecular bases of odor discrimination: Reconstitution of olfactory receptors that recognize overlapping sets of odorants. AB - The vertebrate olfactory system discriminates a wide variety of odorants by relaying coded information from olfactory sensory neurons in the olfactory epithelium to olfactory cortical areas of the brain. Recent studies have shown that the first step in odor discrimination is mediated by approximately 1000 distinct olfactory receptors, which comprise the largest family of G-protein coupled receptors. In the present study, we used Ca(2+) imaging and single-cell reverse transcription-PCR techniques to identify mouse olfactory neurons responding to an odorant and subsequently to clone a receptor gene from the responsive cell. The functionally cloned receptors were expressed in heterologous systems, demonstrating that structurally related olfactory receptors recognized overlapping sets of odorants with distinct affinities and specificities. Our results provide direct evidence for the existence of a receptor code in which the identities of different odorants are specified by distinct combinations of odorant receptors that possess unique molecular receptive ranges. We further demonstrate that the receptor code for an odorant changes with odorant concentration. Finally, we show that odorant receptors in human embryonic kidney 293 cells couple to stimulatory G-proteins such as Galphaolf, resulting in odorant-dependent increases in cAMP. Odor discrimination is thus determined by differences in the receptive ranges of the odorant receptors that together encode specific odorant molecules. PMID- 11487626 TI - Attenuated neurodegenerative disease phenotype in tau transgenic mouse lacking neurofilaments. AB - Previous studies have shown that transgenic (Tg) mice overexpressing human tau protein develop filamentous tau aggregates in the CNS. The most abundant tau aggregates are found in spinal cord and brainstem in which they colocalize with neurofilaments (NFs) as spheroids in axons. To elucidate the role of NF subunit proteins in tau aggregate formation and to test the hypothesis that NFs are pathological chaperones in the formation of intraneuronal tau inclusions, we crossbred previously described tau (T44) Tg mice overexpressing the smallest human tau isoform with knock-out mice devoid of NFL (NFL-/-) or NFH (NFH-/-). Depletion of NF subunit proteins from the T44 mice (i.e., T44;NFL-/- and T44;NFH /-), in particular NFL, resulted in a dramatic decrease in the total number of tau-positive spheroids in spinal cord and brainstem. Concomitant with the reduction in spheroid number, the bigenic mice showed delayed accumulation of insoluble tau protein in the CNS, increased viability, reduced weight loss, and improved behavioral phenotype when compared with the single T44 Tg mice. These results imply that NFs are pathological chaperones in the development of tau spheroids and suggest a role for NFs in the pathogenesis of neurofibrillary tau lesions in neurodegenerative disorders that contain both NFs and tau proteins. PMID- 11487627 TI - Visual transmission deficits in mice with targeted disruption of the gap junction gene connexin36. AB - In the mammalian retina, rods feed into the cone pathway through electrotonic coupling, and recent histological data suggest the involvement of connexin36 (Cx36) in this pathway. We therefore generated Cx36 null mice and monitored the functional consequences of this deficiency on early visual transmission. The homozygous mutant mice had a normally developed retina and showed no changes in the cellular organization of the rod pathway. In contrast, the functional coupling between AII amacrine cells and bipolar cells was impaired. Recordings of electroretinograms revealed a significant decrease of the scotopic b-wave in mutant animals and an increased cone threshold that is compatible with a distorted, gap junctional transmission between AII amacrine cells and cone bipolar cells. Recordings of visual evoked potentials showed extended latency in mutant mice but unaffected ON and OFF components. Our results demonstrate that Cx36-containing gap junctions are essential for normal synaptic transmission within the rod pathway. PMID- 11487628 TI - IPSC kinetics at identified GABAergic and mixed GABAergic and glycinergic synapses onto cerebellar Golgi cells. AB - In the rat cerebellum, Golgi cells receive serotonin-evoked inputs from Lugaro cells (L-IPSCs), in addition to spontaneous inhibitory inputs (S-IPSCs). In the present study, we analyze the pharmacology of these IPSCs and show that S-IPSCs are purely GABAergic events occurring at basket and stellate cell synapses, whereas L-IPSCs are mediated by GABA and glycine. Corelease of the two transmitters at Lugaro cell synapses is suggested by the fact that both GABA(A) and glycine receptors open during individual L-IPSCs. Double immunocytochemical stainings demonstrate that GABAergic and glycinergic markers are coexpressed in Lugaro cell axonal varicosities, together with the mixed vesicular inhibitory amino acid transporter. Lugaro cell varicosities are found apposed to glycine receptor (GlyR) clusters that are localized on Golgi cell dendrites and participate in postsynaptic complexes containing GABA(A) receptors (GABA(A)Rs) and the anchoring protein gephyrin. GABA(A)R and GlyR/gephyrin appear to form segregated clusters within individual postsynaptic loci. Basket and stellate cell varicosities do not face GlyR clusters. For the first time the characteristics of GABA and glycine cotransmission are compared with those of GABAergic transmission at identified inhibitory synapses converging onto the same postsynaptic neuron. The ratio of the decay times of L-IPSCs and of S-IPSCs is a constant value among Golgi cells. This indicates that, despite a high cell-to-cell variability of the overall IPSC decay kinetics, postsynaptic Golgi cells coregulate the kinetics of their two main inhibitory inputs. The glycinergic component of L-IPSCs is responsible for their slower decay, suggesting that glycinergic transmission plays a role in tuning the IPSC kinetics in neuronal networks. PMID- 11487629 TI - Activation of metabotropic glutamate receptor 1 accelerates NMDA receptor trafficking. AB - Regulation of neuronal NMDA receptors (NMDARs) by group I metabotropic glutamate receptors (mGluRs) is known to play a critical role in synaptic transmission. The molecular mechanisms underlying mGluR1-mediated potentiation of NMDARs are as yet unclear. The present study shows that in Xenopus oocytes expressing recombinant receptors, activation of mGluR1 potentiates NMDA channel activity by recruitment of new channels to the plasma membrane via regulated exocytosis. Activation of mGluR1alpha induced (1) an increase in channel number times channel open probability, with no change in mean open time, unitary conductance, or reversal potential; (2) an increase in charge transfer in the presence of NMDA and the open channel blocker MK-801, indicating an increased number of functional NMDARs in the cell membrane; and (3) increased NR1 surface expression, as indicated by cell surface Western blots and immunofluorescence. Botulinum neurotoxin A or expression of a dominant negative mutant of synaptosomal associated protein of 25 kDa molelcular mass (SNAP-25) greatly reduced mGluR1alpha-mediated potentiation, indicating that receptor trafficking occurs via a SNAP-25-mediated form of soluble N-ethylmaleimide sensitive fusion protein attachment protein receptor dependent exocytosis. Because group I mGluRs are localized to the perisynaptic region in juxtaposition to synaptic NMDARs at glutamatergic synapses in the hippocampus, mGluR-mediated insertion of NMDARs may play a role in synaptic transmission and plasticity, including long-term potentiation. PMID- 11487630 TI - The activation of dopamine D4 receptors inhibits oxidative stress-induced nerve cell death. AB - Oxidative stress is thought to be the cause of nerve cell death in many CNS pathologies, including ischemia, trauma, and neurodegenerative disease. Glutamate kills nerve cells that lack ionotropic glutamate receptors via the inhibition of the cystine-glutamate antiporter x(c)(-), resulting in the inhibition of cystine uptake, the loss of glutathione, and the initiation of an oxidative stress cell death pathway. A number of catecholamines were found to block this pathway. Specifically, dopamine and related ligands inhibit glutamate-induced cell death in both clonal nerve cell lines and rat cortical neurons. The protective effects of dopamine, apomorphine, and apocodeine, but not epinephrine and norepinephrine, are antagonized by dopamine D4 antagonists. A dopamine D4 agonist also protects, and this protective effect is inhibited by U101958, a dopamine D4 antagonist. Although the protective effects of some of the catecholamines are correlated with their antioxidant activities, there is no correlation between the protective and antioxidant activities of several other ligands. Normally, glutamate causes an increase in reactive oxygen species (ROS) and intracellular Ca(2+). Apomorphine partially inhibits glutamate-induced ROS production and blocks the opening of cGMP-operated Ca(2+) channels that lead to Ca(2+) elevation in the late part of the cell death pathway. These data suggest that the protective effects of apomorphine on oxidative stress-induced cell death are, at least in part, mediated by dopamine D4 receptors via the regulation of cGMP-operated Ca(2+) channels. PMID- 11487631 TI - Developmental expression of the TTX-resistant voltage-gated sodium channels Nav1.8 (SNS) and Nav1.9 (SNS2) in primary sensory neurons. AB - The development of neuronal excitability involves the coordinated expression of different voltage-gated ion channels. We have characterized the expression of two sensory neuron-specific tetrodotoxin-resistant sodium channel alpha subunits, Na(v)1. (SNS/PN3) and Na(v)1.9 (SNS2/NaN), in developing rat lumbar dorsal root ganglia (DRGs). Expression of both Na(v)1.8 and Na(v)1.9 increases with age, beginning at embryonic day (E) 15 and E17, respectively, and reaching adult levels by postnatal day 7. Their distribution is restricted mainly to those subpopulations of primary sensory neurons in developing and adult DRGs that give rise to unmyelinated C-fibers (neurofilament 200 negative). Na(v)1.8 is expressed in a higher proportion of neuronal profiles than Na(v)1.9 at all stages during development, as in the adult. At E17, almost all Na(v)1.8-expressing neurons also express the high-affinity NGF receptor TrkA, and only a small proportion bind to IB4, a marker for c-ret-expressing (glial-derived neurotrophic factor-responsive) neurons. Because IB4 binding neurons differentiate from TrkA neurons in the postnatal period, the proportion of Na(v)1.8 cells that bind to IB4 increases, in parallel with a decrease in the proportion of Na(v)1.8-TrkA co-expressing cells. In contrast, an equal number of Na(v)1.9 cells bind IB4 and TrkA in embryonic life. The differential expression of Na(v)1.8 and Na(v)1.9 in late embryonic development, with their distinctive kinetic properties, may contribute to the development of spontaneous and stimulus-evoked excitability in small diameter primary sensory neurons in the perinatal period and the activity-dependent changes in differentiation they produce. PMID- 11487632 TI - Cyclin-dependent kinase 4 and cyclin D1 are required for excitotoxin-induced neuronal cell death in vivo. AB - Systemic administration of the glutamic acid analog kainic acid (KA) causes neuronal cell death in brain-vulnerable regions, such as the piriform cortex, hippocampus, and amygdala in rats. We investigated the relationship between the KA-induced neuronal apoptosis and expression of cyclin-dependent kinase 4 (CDK4) and cyclin D1, key regulators of cell cycle progression. Expression of CDK4 and cyclin D1 was upregulated in neurons of the rat piriform cortex and amygdala 1-3 d after KA administration in vivo. CDK4 and cyclin D1 proteins were induced in the cytoplasm and nuclei of neurons, with a concomitant increase of CDK4- and cyclin D1-positive microglia in the affected areas. Continuous infusion of 100 microm CDK4 or cyclin D1 antisense oligonucleotides into the lateral ventricle using mini-osmotic pumps suppressed the excitotoxin-induced neuronal cell death in the piriform cortex and basolateral amygdaloid nucleus, whereas sense oligonucleotides exhibited no such effect. Although KA administration causes prolonged c-Fos expression in the vulnerable regions that preceded the induction of neuronal apoptosis, the CDK4 or cyclin D1 antisense oligonucleotides exhibited no suppressive effect on c-Fos levels. Our results suggest that CDK4 and cyclin D1 are essential for KA-induced neuronal apoptosis in vivo. PMID- 11487633 TI - Ducky mouse phenotype of epilepsy and ataxia is associated with mutations in the Cacna2d2 gene and decreased calcium channel current in cerebellar Purkinje cells. AB - The mouse mutant ducky, a model for absence epilepsy, is characterized by spike wave seizures and ataxia. The ducky gene was mapped previously to distal mouse chromosome 9. High-resolution genetic and physical mapping has resulted in the identification of the Cacna2d2 gene encoding the alpha2delta2 voltage-dependent calcium channel subunit. Mutations in Cacna2d2 were found to underlie the ducky phenotype in the original ducky (du) strain and in a newly identified strain (du(2J)). Both mutations are predicted to result in loss of the full-length alpha2delta2 protein. Functional analysis shows that the alpha2delta2 subunit increases the maximum conductance of the alpha1A/beta4 channel combination when coexpressed in vitro in Xenopus oocytes. The Ca(2+) channel current in acutely dissociated du/du cerebellar Purkinje cells was reduced, with no change in single channel conductance. In contrast, no effect on Ca(2+) channel current was seen in cerebellar granule cells, results consistent with the high level of expression of the Cacna2d2 gene in Purkinje, but not granule, neurons. Our observations document the first mammalian alpha2delta mutation and complete the association of each of the major classes of voltage-dependent Ca(2+) channel subunits with a phenotype of ataxia and epilepsy in the mouse. PMID- 11487634 TI - Spine formation and correlated assembly of presynaptic and postsynaptic molecules. AB - Hippocampal pyramidal neurons in culture showed a developmental shift in synapse distribution from dendritic shafts to spines. Using dual wavelength time-lapse fluorescence microscopy, we analyzed the morphogenesis of three synaptic components: dendritic spines, postsynaptic densities (PSDs), and presynaptic vesicles. Local assembly of a major PSD protein, PSD-95, was spatially and temporally correlated with spine morphogenesis. Clustering of postsynaptic PSD-95 and that of a predominant synaptic vesicle protein, synaptophysin, were also correlated. In contrast, pre-existing PSD-95 clusters in dendritic shafts were preferentially eliminated without promoting spine formation. The local and stepwise assembly of synaptic components at the contact sites between dendritic protrusions and axons explains the developmental remodeling of excitatory synapses. PMID- 11487635 TI - Regulation of dendritic spine motility in cultured hippocampal neurons. AB - Regulation of dendritic spine motility was studied in dissociated cultures of the rat and mouse hippocampus, using green fluorescent protein-labeled neurons or neurons loaded with the calcium-sensitive dye Oregon Green-1. Cells were time lapse-photographed on a confocal laser-scanning microscope at high resolution to detect movements as well as spontaneous fluctuations of intracellular calcium concentrations in their dendritic spines. Active presynaptic terminals attached to the spines were labeled with FM4-64, which marks a subset of synaptophysin labeled terminals. Dendritic spines were highly motile in young, 4- to 7-d-old cells. At this age, neurons had little spontaneous calcium fluctuation or FM4-64 labeling. Within 2-3 weeks in culture, dendritic spines were much less motile, they were associated with active presynaptic terminals, and they expressed high rates of spontaneous calcium fluctuations. Irrespective of age, and even on the same dendrite, there was an inverse relationship between spine motility and presence of FM4-64-labeled terminals in contact with the imaged spines. Spine motility was blocked by latrunculin, which prevents actin polymerization, and was disinhibited by blockade of action potential discharges with tetrodotoxin. It is proposed that an active presynaptic terminal restricts motility of dendritic spines. PMID- 11487636 TI - Schwann cell type V collagen inhibits axonal outgrowth and promotes Schwann cell migration via distinct adhesive activities of the collagen and noncollagen domains. AB - Previously, we reported the cloning of alpha4 type V collagen, a novel member of the collagen type V gene family that is expressed by Schwann cells in developing peripheral nerves (Chernousov et al., 2000). The present study was performed to investigate the effects of this collagen on the adhesion and migration of premyelinating Schwann cells and neurite outgrowth from embryonic dorsal root ganglion neurons. Purified alpha4(V)-containing collagen isolated from Schwann cell conditioned medium (collagen type V(SC)) promoted migration of Schwann cells but inhibited outgrowth of axons from rat embryo dorsal root ganglia. Collagen type V(SC) blocked axonal outgrowth in the presence of otherwise active substrates such as collagen type IV, indicative of active inhibition. The noncollagen N-terminal domain of alpha4(V) promoted Schwann cell adhesion, spreading, and migration. These processes were inhibited by soluble heparin but not by function-blocking antibodies against alpha1- and alpha2-integrins. The collagen domain of pepsin-digested collagen type V was poorly adhesive for Schwann cells. The type V collagen domain but not the alpha4(V) N-terminal domain blocked neurite outgrowth from dorsal root ganglion neurons. In cocultures of dorsal root ganglion neurons and Schwann cells, collagen type V(SC) promoted axon fasciculation and association of axons with Schwann cells. These results suggest that in embryonic peripheral nerves, collagen type V(SC) plays a dual role in regulating cell migration. This represents a heretofore unrecognized function of peripheral nerve collagen fibrils in regulating patterns of peripheral nerve growth during development. PMID- 11487637 TI - Glial cell line-derived neurotrophic factor administration in postnatal life results in motor unit enlargement and continuous synaptic remodeling at the neuromuscular junction. AB - Overexpression of glial cell line-derived neurotrophic factor (GDNF) in embryonic muscle fibers causes dramatic hyperinnervation of neuromuscular junctions. However, it is not known whether GDNF induces the extra innervation by regulation of axonal branching and/or synaptic maintenance. To address this issue, high levels of circulating GDNF were established by administering subcutaneous injections starting either at birth or later and continuing for up to 40 d. Treatment with exogenous GDNF beginning in the first week, but not later, increased the number of axons converging at neuromuscular junctions. The effect of GDNF on the branching pattern of individual motor axons was determined by reconstructing labeled axonal arbors from transgenic mice expressing yellow fluorescent protein in subsets of motor neurons. Whereas, at postnatal day 8 (P8) individual axons in control animals branched to sporadically innervate junctions within circumscribed regions of the muscle, motor units from GDNF injected animals had significantly more axonal branches and exhibited a high degree of localized arborization such that adjacent muscle fibers were often innervated by the same axon. Administration beginning at P0 and continuing through P40 prolonged multiple innervation of most fibers throughout the period of injection. Between P30 and P40 there was no net change in multiple innervation, although there was evidence of retraction bulbs, suggesting that axon extension and retraction were in equilibrium. We conclude that GDNF has a developmentally regulated effect on presynaptic branching and that sustained administration of GDNF induces a state of continuous synaptic remodeling. PMID- 11487638 TI - Inactivation of the glial fibrillary acidic protein gene, but not that of vimentin, improves neuronal survival and neurite growth by modifying adhesion molecule expression. AB - Intermediate filaments (IFs) are a major component of the cytoskeleton in astrocytes. Their role is far from being completely understood. Immature astrocytes play a major role in neuronal migration and neuritogenesis, and their IFs are mainly composed of vimentin. In mature differentiated astrocytes, vimentin is replaced by the IF protein glial fibrillary acidic protein (GFAP). In response to injury of the CNS in the adult, astrocytes become reactive, upregulate the expression of GFAP, and reexpress vimentin. These modifications contribute to the formation of a glial scar that is obstructive to axonal regeneration. Nevertheless, astrocytes in vitro are considered to be the ideal substratum for the growth of embryonic CNS axons. In the present study, we have examined the potential role of these two major IF proteins in both neuronal survival and neurite growth. For this purpose, we cocultured wild-type neurons on astrocytes from three types of knock-out (KO) mice for GFAP or/and vimentin in a neuron-astrocyte coculture model. We show that the double KO astrocytes present many features of immaturity and greatly improve survival and neurite growth of cocultured neurons by increasing cell-cell contact and secreting diffusible factors. Moreover, our data suggest that the absence of vimentin is not a key element in the permissivity of the mutant astrocytes. Finally, we show that only the absence of GFAP is associated with an increased expression of some extracellular matrix and adhesion molecules. To conclude, our results suggest that GFAP expression is able to modulate key biochemical properties of astrocytes that are implicated in their permissivity. PMID- 11487639 TI - Myosin IIB is required for growth cone motility. AB - Growth cones are required for the forward advancement and navigation of growing axons. Modulation of growth cone shape and reorientation of the neurite are responsible for the change of outgrowth direction that underlies navigation. Change of shape involves the reordering of the cytoskeleton. Reorientation of the neurite requires the generation of tension, which is supplied by the ability of the growth cone to crawl on a substrate. The specific molecular mechanisms responsible for these activities are unknown but are thought to involve actomyosin-generated force combined with linkage to the cell surface receptors that are responsible for adhesion (Heidemann and Buxbaum, 1998). To test whether myosin IIB is responsible for the force generation, we quantified shape dynamics and filopodial-mediated traction force in growth cones from myosin IIB knock-out (KO) mice and compared them with neurons from normal littermates. Growth cones from the KO mice spread less, showed alterations in shape dynamics and actin organization, and had reduced filopodial-mediated traction force. Although peak traction forces produced by filopodia of KO cones were decreased significantly, KO filopodia occasionally developed forces equivalent to those in the wild type. This indicates that other myosins participate in filopodial-dependent traction force. Therefore, myosin IIB is necessary for normal growth cone spreading and the modulation of shape and traction force but acts in combination with other myosins for some or all of these activities. These activities are essential for growth cone forward advancement, which is necessary for outgrowth. Thus outgrowth is slowed, but not eliminated, in neurons from the myosin IIB KO mice. PMID- 11487640 TI - Spatial shaping of cochlear innervation by temporally regulated neurotrophin expression. AB - Previous work suggested qualitatively different effects of neurotrophin 3 (NT-3) in cochlear innervation patterning in different null mutants. We now show that all NT-3 null mutants have a similar phenotype and lose all neurons in the basal turn of the cochlea. To understand these longitudinal deficits in neurotrophin mutants, we have compared the development of the deficit in the NT-3 mutant to the spatial-temporal expression patterns of brain-derived neurotrophic factor (BDNF) and NT-3, using lacZ reporters in each gene and with expression of the specific neurotrophin receptors, trkB and trkC. In the NT-3 mutant, almost normal numbers of spiral ganglion neurons form, but fiber outgrowth to the basal turn is eliminated by embryonic day (E) 13.5. Most neurons are lost between E13.5 and E15.5. During the period preceding apoptosis, NT-3 is expressed in supporting cells, whereas BDNF is expressed mainly in hair cells, which become postmitotic in an apical to basal temporal gradient. During the period of neuronal loss, BDNF is absent from the basal cochlea, accounting for the complete loss of basal turn neurons in the NT-3 mutant. The spatial gradients of neuronal loss in these two mutants appear attributable to spatial-temporal gradients of neurotrophin expression. Our immunocytochemical data show equal expression of their receptors, TrkB and TrkC, in spiral sensory neurons and thus do not relate to the basal turn loss. Mice in which NT-3 was replaced by BDNF show a qualitative normal pattern of innervation at E13.5. This suggests that the pattern of expression of neurotrophins rather than their receptors is essential for the spatial loss of spiral sensory neurons in NT-3 null mutants. PMID- 11487641 TI - Cyclic nucleotide-mediated regulation of hippocampal mossy fiber development: a target-specific guidance. AB - The mossy fibers (MFs) arising from dentate granule cells project primarily onto a narrow segment of the proximal dendrites of hippocampal CA3 pyramidal cells. The mechanisms underlying this specific MF target selection are not fully understood. To investigate the cellular basis for development of the stereotyped MF trajectories, we have arranged the fascia dentata and hippocampal Ammon's horn tissues in diverse topographical patterns in organotypic explant coculture systems. Here we show that cyclic nucleotide signaling pathways regulate the MF pathfinding. When the dentate gyrus explants were ectopically placed facing the CA3 stratum oriens of hippocampal slices, MFs crossed the border between cocultures and reached their appropriate target area in the Ammon's horn, as assessed by membrane tracer labeling, Timm staining, electrophysiological recording of synaptic responses, and optical analyses using a voltage-sensitive dye. This lamina-specific MF innervation was disrupted by pharmacological blockade of cGMP pathway. Similar apposition of the dentate grafts near the CA1 region of host slices rarely resulted in MF ingrowth into the Ammon's horn. Under blockade of cAMP pathway, however, the MFs were capable of making allopatric synapses with CA1 neurons. These data were further supported by the pharmacological data obtained from granule cells dispersed over hippocampal slice cultures. Thus, our findings suggest that the stereotyped MF extension is mediated by at least two distinct factors, i.e., an attractant derived from the CA3 region and a repellent from the CA1 region. These factors may be regulated differently by cAMP and cGMP signaling pathways. PMID- 11487642 TI - Aldolase C/zebrin II expression in the neonatal rat forebrain reveals cellular heterogeneity within the subventricular zone and early astrocyte differentiation. AB - During late gestational and early postnatal development, proliferating cells in the subventricular zones of the lateral ventricles (SVZ) migrate into the gray and white matter of the forebrain and differentiate into astrocytes and oligodendrocytes. Because the cellular composition and structure of the neonatal SVZ is poorly understood, we performed a differential display PCR screen to identify genes preferentially expressed therein. One highly expressed gene encoded aldolase C. We used a specific monoclonal antibody, aldolase C/zebrin II (ALDC/ZII), in combination with markers of glial lineage and proliferation, to characterize the cells that express this gene. In the neonatal SVZ, ALDC/ZII positive cells, which are generally polygonal and display several processes, have a nonuniform spatial distribution. They do not express vimentin, GFAP, or NG2. A subset of ALDC/ZII-positive cells incorporates bromodeoxyuridine, but progenitors identified by beta-galactosidase expression after infection with recombinant BAG virus do not show ALDC/ZII immunoreactivity. Outside of the SVZ, beta galactosidase-positive/ALDC/ZII-positive cells have an astrocytic phenotype, suggesting that immunoreactivity was acquired after exit from the SVZ. These studies demonstrate that the neonatal SVZ is composed of different populations of cells that can be characterized by their antigenic phenotype, their proliferative capacity, and their spatial distributions. Nonrandom distributions of different cell types within the SVZ may permit the formation of microenvironments that stimulate the production of cells with specific potentials at appropriate points in development. Analysis of ALDC/ZII expression by astrocyte lineage cells in the neonatal cerebral cortex and white matter may reveal insights into the phenotype and behavior of undifferentiated astrocyte progenitors. PMID- 11487643 TI - Axonal regeneration into acellular nerve grafts is enhanced by degradation of chondroitin sulfate proteoglycan. AB - Although the peripheral nerve has the potential to regenerate after injury, degenerative processes may be essential to promote axonal growth into the denervated nerve. One hypothesis is that the nerve contains growth inhibitors that must be neutralized after injury for optimal regeneration. In the present study, we tested whether degradation of chondroitin sulfate proteoglycan, a known inhibitor of axon growth, enhances the growth-promoting properties of grafts prepared from normal donor nerves. Excised segments of rat sciatic nerve were made acellular by freeze-killing before treatment with chondroitinase ABC. Chondroitinase-dependent neoepitope immunolabeling showed that chondroitin sulfate proteoglycan was thoroughly degraded throughout the treated nerve segments. In addition, neuronal cryoculture assays revealed that the neurite promoting activity of acellular nerves was significantly increased by chondroitinase treatment. Control and chondroitinase-treated acellular nerves were then used as interpositional grafts in a rat nerve injury model. Axonal regeneration into the grafts was assessed 4 and 8 d after implantation by growth associated protein-43 immunolabeling. At both time points, the number of axons regenerating into acellular grafts treated with chondroitinase was severalfold greater than in control grafts. Growth into the chondroitinase-treated grafts was pronounced after only 4 d, suggesting that the delay of axonal growth normally associated with acellular grafts was attenuated as well. These findings indicate that chondroitinase treatment significantly enhanced the growth-promoting properties of freeze-killed donor nerve grafts. Combined with the low immunogenicity of acellular grafts, the ability to improve axonal penetration into interpositional grafts by preoperative treatment with chondroitinase may be a significant advancement for clinical nerve allografting. PMID- 11487644 TI - Acute neuronal apoptosis in a rat model of multiple sclerosis. AB - Demyelination caused by inflammation of the CNS has been considered to be a major hallmark of multiple sclerosis (MS). Using experimental autoimmune encephalomyelitis, a model of MS, we demonstrate that an immune-mediated attack of the optic nerve is accompanied by an early degeneration of retinal ganglion cells (RGCs). The decrease of neuronal cell density was correlated with functional disabilities as assessed by visual evoked cortical potentials and electroretinogram. Visual acuity was significantly reduced. DNA degradation and activation of caspase-3 in RGCs indicate that cell death of RGCs is apoptotic. These findings show for the first time that an inflammatory attack against myelin components can lead to acute neuronal cell loss by apoptosis. PMID- 11487645 TI - Effects of progesterone synthesized de novo in the developing Purkinje cell on its dendritic growth and synaptogenesis. AB - De novo steroidogenesis from cholesterol is a conserved property of vertebrate brains, and such steroids synthesized de novo in the brain are called neurosteroids. The identification of neurosteroidogenic cells is essential to the understanding of the physiological role of neurosteroids in the brain. We have demonstrated recently that neuronal neurosteroidogenesis occurs in the brain and indicated that the Purkinje cell actively synthesizes several neurosteroids de novo from cholesterol in vertebrates. Interestingly, in the rat, this neuron actively synthesizes progesterone de novo from cholesterol only during neonatal life, when cerebellar cortical formation occurs most markedly. Therefore, in this study, the possible organizing actions of progesterone during cerebellar development have been examined. In vitro studies using cerebellar slice cultures from newborn rats showed that progesterone promotes dose-dependent dendritic outgrowth of Purkinje cells but dose not affect their somata. This effect was blocked by the anti-progestin RU 486 [mifepristone; 17beta-hydroxy-11beta-(4 methylaminophenyl)-17alpha-(1-propynyl) estra-4,9-dien-3 one-6-7]. In vivo administration of progesterone to pups further revealed an increase in the density of Purkinje spine synapses electron microscopically. In contrast to progesterone, there was no significant effect of 3alpha,5alpha tetrahydroprogesterone, a progesterone metabolite, on Purkinje cell development. Reverse transcription-PCR-Southern and immunocytochemical analyses showed that intranuclear progesterone receptors were expressed in Purkinje cells. These results suggest that progesterone promotes both dendritic outgrowth and synaptogenesis in Purkinje cells through intranuclear receptor-mediated mechanisms during cerebellar development. Such organizing actions may contribute to the formation of the cerebellar neuronal circuit. PMID- 11487646 TI - Developmental depression of glutamate neurotransmission by chronic low-level activation of NMDA receptors. AB - Slabs of slow-release plastic (Elvax) containing NMDA or solvent were implanted over the rat colliculus beginning on postnatal day 8 (P8). Whole-cell patch clamping in the superficial superior collicular layers (sSCs) from P10 to P21 demonstrated a severe decrease in spontaneous EPSC frequency after chronic NMDA treatment. The decrease was not attributable to an increase in GABA(A) receptor mediated inhibition and was present only when NMDA receptor (NMDAR) current was blocked by Mg(2+). Analysis of miniature EPSCs indicated that many active sites on NMDA-treated neurons lacked functional AMPA and kainate receptor (AMPA/KAR) currents, and AMPA/KAR:NMDAR current ratios of evoked EPSCs were also significantly reduced. In addition, the normal downregulation of NMDAR decay time in sSC neurons at P11 was absent after NMDA treatment. Nevertheless, neither AMPA nor NMDA receptor subunit expression was altered by NMDA treatment, and experiments with the NMDAR antagonist ifenprodil suggested that incorporation of NR2A-containing NMDARs at the sSC synapses was unperturbed. Thus, disrupting but not blocking NMDARs suppresses the development of AMPA/KAR currents. The absence of the P11 NMDAR current downregulation is likely a secondary effect resulting from the reduction of AMPA/KAR function. Chronic agonist application reduces but does not eliminate NMDAR conductances. Therefore these data support an active role for NMDAR currents in synaptic development. Prolonged NMDA treatment in vivo, which couples reduced postsynaptic Ca(2+) responses with normally developing afferent activity, produces a long-lasting synaptic depression and stalls glutamatergic synaptogenesis, suggesting that the correlation between robust NMDAR activation and afferent activity is an essential component during normal development. PMID- 11487647 TI - Remodeling of synaptic membranes after induction of long-term potentiation. AB - Several morphological changes of synapses have been reported to be associated with the induction of long-term potentiation (LTP) in the CA1 hippocampus, including an transient increase in the proportion of synapses with perforated postsynaptic densities (PSDs) and a later occurrence of multiple spine boutons (MSBs) in which the two spines arise from the same dendrite. To investigate the functional significance of these modifications, we analyzed single sections and reconstructed 134 synapses labeled via activity using a calcium precipitation approach. Analyses of labeled spine profiles showed changes of the spine head area, PSD length, and proportion of spine profiles containing a coated vesicle that reflected variations in the relative proportion of different types of synapses. Three-dimensional reconstruction indicated that the increase of perforated spine profiles observed 30 min after LTP induction essentially resulted from synapses exhibiting segmented, completely partitioned PSDs. These synapses had spine head and PSD areas approximately three times larger than those of simple synapses. They contained coated vesicles in a much higher proportion than that of any other type of synapse and exhibited large spinules associated with the PSD. Also the MSBs with two spines arising from the same dendrite that were observed 1-2 hr after LTP induction included a spine that was smaller and a PSD that was smaller than those of simple synapses. These results support the idea that LTP induction is associated with an enhanced recycling of synaptic membrane and that this process could underlie the formation of synapses with segmented PSDs and eventually result in the formation of a new, immature spine. PMID- 11487648 TI - Restorative plasticity of dopamine neuronal transplants depends on the degree of hemispheric dominance. AB - The ability of dopaminergic (DA) transplants to restore complex sensorimotor behaviors in experimental Parkinson's disease is dependent on graft survival and reinnervation and is likely to be further modified by complex functional graft host interactions. Here, we examined the impact of hemispheric dominance and extensive testing regimes on the functional capabilities of DA transplants to restore skilled forelimb movements in rats with unilateral 6-hydroxydopamine lesions. Interestingly, a near complete recovery was observed in DA-grafted animals that did not exhibit a strong hemispheric lateralization for paw use before lesion and implantation surgery, whereas animals with a clear lateralization of paw use and grafted into the contralateral hemisphere exhibited only moderate recovery. Finally, animals grafted ipsilateral to the preferred paw were most resistant to functional improvements in skilled forelimb use. However, the influence of hemispheric dominance on the degree of functional DA graft induced restoration was specific for skilled forelimb use, whereas no such differences were observed in other tests for motor and sensory functions related to the DA system. Furthermore, functional recovery of DA-grafted animals in skilled forelimb use was significantly promoted by extensive behavioral testing regimes indicative of a "learning how to use" the transplant effect. These findings indicate the importance of the underlying functional architecture of complex sensorimotor behaviors, such as skilled forelimb use, and the DA neurotransmitter system for the plasticity of DA transplants to promoting a more complete behavioral recovery in experimental, and potentially, also in clinical forms of Parkinson's disease. PMID- 11487649 TI - Increasing the number of synapses modifies olfactory perception in Drosophila. AB - The Drosophila mutant gigas produces an enlargement of postmitotic cells caused by additional rounds of DNA replication. In neurons, the mutant cell establishes more synapses than normal. We have taken advantage of this feature to study the effect of synapse number on odorant perception. Mosaic adults were generated in which one antenna was homozygous for gigas, whereas the contralateral side served as an internal control. Morphological analysis indicates that the number and type of sensory afferents forming the mutant antenna, as well as their projection to the olfactory glomeruli, are normal. In contrast, the volume of identified glomeruli increases to a variable extent, and mutant sensory neurons branch profusely. The number of synapses, estimated in the ventral (V) glomerulus that receives ipsilateral afferents only, is increased twofold to threefold. Large dense-core vesicle-containing terminals that probably modulate olfactory centers are identified in the V glomerulus. Their number and size are not modified by the mutant input. Sensory transduction, measured by electroantennograms, is normal in amplitude and kinetics. In odorant tests, however, the profile of the behavioral response to ethyl acetate shows attractive responses to concentrations to which sibling controls remain indifferent (10(-)8 and 10(-)7 v/v). In addition, the intensity of the response is augmented both at attractive and repulsive odorant concentrations with respect to that of controls. These results demonstrate that increased synapse number in the sensory neurons can modify the behavior of the organism, allowing a higher sensitivity of perception. PMID- 11487650 TI - Odor exposure causes central adaptation and morphological changes in selected olfactory glomeruli in Drosophila. AB - In an attempt to correlate behavioral and neuronal changes, we examined the structural and functional effects of odor exposure in Drosophila. Young adult flies were exposed to a high concentration of the selected odor, usually benzaldehyde or isoamyl acetate, for 4 d and subsequently tested for their olfactory response to a variety of odorants and concentrations. The behavioral response showed specific adaptation to the exposed odor. By contrast, olfactory transduction, as measured in electroantennograms, remained normal. In vivo volume measurements were performed on olfactory glomeruli, the anatomical and functional units involved in odor processing. Pre-exposed flies exhibited volume reduction of certain glomeruli, in an odor-selective manner. Of a sample of eight glomeruli measured, dorsal medial (DM) 2 and ventral (V) were affected by benzaldehyde exposure, whereas DM6 was affected by isoamyl acetate. Estimation of the number of synapses indicates that volume reduction involves synapse loss that can reach 30% in the V glomerulus of flies adapted to benzaldehyde. Additional features of odorant-induced adaptation, including concentration dependence and perdurance, also show correlation, because both effects are elicited by high odor concentrations and are long-lasting (>1 week). Finally, the dunce mutant fails to develop behavioral adaptation as well as morphological changes in the olfactory glomeruli after exposure. These neural changes thus appear to require the cAMP signaling pathway. PMID- 11487651 TI - The inferior parietal lobule is the target of output from the superior colliculus, hippocampus, and cerebellum. AB - The inferior parietal lobule (IPL) is a functionally and anatomically heterogeneous region that is concerned with multiple aspects of sensory processing and sensorimotor integration. Although considerable information is available about the corticocortical connections to the IPL, much less is known about the origin and importance of subcortical inputs to this cortical region. To examine this issue, we used retrograde transneuronal transport of the McIntyre-B strain of herpes simplex virus type 1 (HSV1) to identify the second-order neurons in subcortical nuclei that project to the IPL. Four monkeys (Cebus apella) received injections of HSV1 into three different subregions of the IPL. Injections into a portion of the lateral intraparietal area labeled second-order neurons primarily in the superficial (visual) layers of the superior colliculus. Injections of HSV1 into a portion of area 7a labeled many second-order neurons in the CA1 region of the hippocampus. In contrast, virus injections within a portion of area 7b labeled second-order neurons in posterior regions of the dentate nucleus of the cerebellum. These observations have some important functional implications. The IPL is known to be involved in oculomotor and attentional mechanisms, the establishment of maps of extrapersonal space, and the adaptive recalibration of eye-hand coordination. Our findings suggest that these functions are subserved by distinct subcortical systems from the superior colliculus, hippocampus, and cerebellum. Furthermore, the finding that each system appears to target a separate subregion of the IPL provides an anatomical substrate for understanding the functional heterogeneity of the IPL. PMID- 11487652 TI - Sex differences and opposite effects of stress on dendritic spine density in the male versus female hippocampus. AB - Dendritic spines are postsynaptic sites of excitatory input in the mammalian nervous system. Despite much information about their structure, their functional significance remains unknown. It has been reported that females in proestrus, when estrogen levels are elevated, have a greater density of apical dendritic spines on pyramidal neurons in area CA1 of the hippocampus than females in other stages of estrous (Woolley et al., 1990). Here we replicate these findings and in addition, show that females in proestrus have a greater density of spines in area CA1 of the hippocampus than males. Moreover, this sex difference in spine density is affected in opposite directions by stressful experience. In response to one acute stressful event of intermittent tailshocks, spine density was enhanced in the male hippocampus but reduced in the female hippocampus. The decrease in the female was observed for those that were stressed during diestrus 2 and perfused 24 hr later during proestrus. The opposing effects of stress were not evident immediately after the stressor but rather occurred within 24 hr and were evident on apical and to a lesser extent on basal dendrites of pyramidal cells in area CA1. Neither sex nor stress affected spine density on pyramidal neurons in somatosensory cortex. Sex differences in hippocampal spine density correlated with sex hormones, estradiol and testosterone, whereas stress effects on spine density were not directly associated with differences in the stress hormones, glucocorticoids. In summary, males and females have different levels of dendritic spine density in the hippocampus under unstressed conditions, and their neuronal anatomy can respond in opposite directions to the same stressful event. PMID- 11487653 TI - Quantitative analysis of synaptic contacts made between functionally identified oralis neurons and trigeminal motoneurons in cats. AB - A previous study revealed that rostrodorsomedial oralis (Vo.r) neurons synapsing on trigeminal motoneurons use GABA and/or glycine as neurotransmitters. To determine the number and spatial distribution of contacts, injections of biotinamide and horseradish peroxidase were made into a Vo.r neuron and an alpha motoneuron in the jaw-closing (JC) and jaw-opening (JO) motor nucleus, respectively, in 39 cats. All Vo.r neurons responded to low-threshold mechanical stimulation of the oral tissues. Single Vo.r neurons terminating in the JC nucleus (Vo.r-dl neurons; n = 5) issued, on average, 10 times more boutons than Vo.r neurons terminating in the JO nucleus (Vo.r-vm neurons; n = 5; 4437 vs 445). The Vo.r-dl neuron-JC alpha-motoneuron pairs (n = 4) made contacts on either the soma-dendritic compartment or dendrites, and the Vo.r-vm neuron-JO motoneuron pairs (n = 2) made contacts on dendrites, with a range of two to seven contacts. In five of the six pairs, individual or groups of two to three terminals contacted different dendritic branches of a postsynaptic cell. The Vo.r-dl neurons innervated a greater number of counter-stained motoneuronal somata than did the Vo.r-vm neurons (216 vs 26). Total number of contacts per Vo.r neuron was higher for the Vo.r-dl than Vo.r-vm neurons (786 vs 72). The present study demonstrates that axonal branches of Vo.r neurons are divided into two types with different innervation domains on the postsynaptic neuron and that they are highly divergent. The overall effect exerted by these neurons is predicted to be much greater within the JC than JO motoneuron pool. PMID- 11487654 TI - Adenosine A1 receptors reduce release from excitatory but not inhibitory synaptic inputs onto lateral horn neurons. AB - Although adenosine is an important neuromodulator in the CNS, its role in modulating sympathetic outflow at the level of the spinal cord has not been studied. Because very little is known about adenosine A1 receptors (A1Rs) in the spinal cord, we determined their location and role with particular reference to the control of sympathetic preganglionic activity and interneuronal activity in the rat. High levels of immunoreactivity for A1Rs were observed throughout the spinal cord. Immunostaining was dense in the intermediolateral cell column (IML) and intercalated nucleus, regions containing retrogradely labeled sympathetic preganglionic neurons (SPNs). Electron microscopy revealed A1R immunoreactivity (A1R-IR) within presynaptic terminals and (to a lesser extent) postsynaptic structures in the IML, as well as the luminal membrane of endothelial cells lining capillaries. Using double-labeling techniques, some presynaptic terminals were observed to synapse onto SPNs. To investigate the effects of activating these A1Rs, visualized whole-cell patch-clamp recordings were made from electrophysiologically and morphologically identified SPNs and interneurons. Applications of the A1R agonist cyclopentyladenosine (CPA) reduced the amplitude of EPSPs elicited by stimulation of the lateral funiculus, an effect blocked by the A1R antagonist 8-cyclopentyl-1,3-dipropylxanthine. These effects were attributable to adenosine acting at a presynaptic site because CPA application increased the paired-pulse ratio. CPA did not affect evoked IPSPs. These data show that activating A1Rs reduces fast excitatory, but not inhibitory, transmission onto SPNs and interneurons in the IML and that A1Rs may play a protective role on neurons involved in the control of sympathetic outflow. PMID- 11487655 TI - Spatial localization after excision of human auditory cortex. AB - Neurophysiological and animal ablation studies concur that primary auditory cortex is necessary for computation of the spatial coordinates of a sound source. Human studies have reported conflicting findings but have often suffered from inadequate psychophysical measures and/or poor lesion localization. We tested patients with unilateral temporal lobe excisions either encroaching on or sparing Heschl's gyrus (HG), quantifying lesion extent using anatomical magnetic resonance imaging measures. Subjects performed two tasks. In the localization task, they heard single clicks in a free-field spatial array subtending 180 degrees of azimuth and indicated the perceived location with a laser pointer. In the discrimination task, two clicks were presented, and subjects indicated if they were in the same or different position. As a group, patients with right temporal excision, either encroaching onto HG or not, were significantly impaired in both hemifields in both tasks, although this was not true for all individuals. Patients with left temporal resections generally performed normally, although some of the patients with left HG excision showed impaired performance bilaterally, especially in the discrimination task. This pattern stands in marked contrast to previous studies showing significant preservation of localization in hemispherectomized patients. We conclude that (1) contrary to hypotheses derived from animal studies, human auditory spatial processes are dependent primarily on cortical areas within right superior temporal cortex, which encompass both spatial hemifields; (2) functional reorganization may not take place after restricted focal damage but only after more extensive early damage; and (3) the existence of individual differences likely illustrates differential patterns of functional lateralization and/or recovery. PMID- 11487656 TI - Long-range synchrony in the gamma band: role in music perception. AB - Synchronization seems to be a central mechanism for neuronal information processing within and between multiple brain areas. Furthermore, synchronization in the gamma band has been shown to play an important role in higher cognitive functions, especially by binding the necessary spatial and temporal information in different cortical areas to build a coherent perception. Specific task-induced (evoked) gamma oscillations have often been taken as an indication of synchrony, but the presence of long-range synchrony cannot be inferred from spectral power in the gamma range. We studied the usefulness of a relatively new measure, called similarity index to detect asymmetric interdependency between two brain regions. Spontaneous EEG from two groups-musicians and non-musicians-were recorded during several states: listening to music, listening to text, and at rest (eyes closed and eyes open). While listening to music, degrees of the gamma band synchrony over distributed cortical areas were found to be significantly higher in musicians than non-musicians. Yet no differences between these two groups were found at resting conditions and while listening to a neutral text. In contrast to the degree of long-range synchrony, spectral power in the gamma band was higher in non-musicians. The degree of spatial synchrony, a measure of signal complexity based on eigen-decomposition method, was also significantly increased in musicians while listening to music. As compared with non-musicians, the finding of increased long-range synchrony in musicians independent of spectral power is interpreted as a manifestation of a more advanced musical memory of musicians in binding together several features of the intrinsic complexity of music in a dynamical way. PMID- 11487657 TI - Preferential increases in nucleus accumbens dopamine after systemic cocaine administration are caused by unique characteristics of dopamine neurotransmission. AB - In vivo voltammetry was used to investigate the preferential increase of extracellular dopamine in the nucleus accumbens relative to the caudate-putamen after systemic cocaine administration. In the first part of this study, cocaine (40 mg/kg, i.p.) was compared with two other blockers of dopamine uptake, nomifensine (10 mg/kg, i.p.) and 3beta-(p-chlorophenyl)tropan-2beta-carboxylic acid p-isothiocyanatophenylmethyl ester hydrochloride (RTI-76; 100 nmol, i.c.v.), to assess whether the inhibitory mechanism of cocaine differed in the two regions. All three drugs robustly increased electrically evoked levels of dopamine, and cocaine elevated dopamine signals to a greater extent in the nucleus accumbens. However, kinetic analysis of the evoked dopamine signals indicated that cocaine and nomifensine increased the K(m) for dopamine uptake whereas the dominant effect of RTI-76 was a decrease in V(max). Under the present in vivo conditions, therefore, cocaine is a competitive inhibitor of dopamine uptake in both the nucleus accumbens and caudate-putamen. Whether the preferential effect of cocaine was mediated by regional differences in the presynaptic control of extracellular DA that are described by rates for DA uptake and release was examined next by a correlation analysis. The lower rates for dopamine release and uptake measured in the nucleus accumbens were found to underlie the preferential increase in extracellular dopamine after cocaine. This relationship explains the paradox that cocaine more effectively increases accumbal dopamine despite identical effects on the dopamine transporter in the two regions. The mechanism proposed for the preferential actions of cocaine may also mediate the differential effects of psychostimulant in extrastriatal regions and other uptake inhibitors in the striatum. PMID- 11487658 TI - Maintenance of serotonin in the intestinal mucosa and ganglia of mice that lack the high-affinity serotonin transporter: Abnormal intestinal motility and the expression of cation transporters. AB - The enteric serotonin reuptake transporter (SERT) has been proposed to play a critical role in serotonergic neurotransmission and in the initiation of peristaltic and secretory reflexes. We analyzed potential compensatory mechanisms and enteric function in the bowels of mice with a targeted deletion of SERT. The guts of these animals were found to lack mRNA encoding SERT; moreover, high affinity uptake of 5-HT into epithelial cells, mast cells, and enteric neurons was present in the SERT +/+ bowel but absent in the SERT -/- bowel. However, both the SERT +/+ gut and the -/- gut expressed molecules capable of transporting 5 HT, but with affinities and selectivity much lower than those of SERT. These included the dopamine transporter (DAT) and polyspecific organic cation transporters OCT-1 and OCT-3. DAT and OCT immunoreactivities were present in both the submucosal and myenteric plexuses, and the OCTs were also located in the mucosal epithelium. 5-HT was found in all of its normal sites in the SERT -/- bowel, which contained mRNA encoding tryptophan hydroxylase, but no 5-HT was present in the blood of SERT -/- animals. Stool water and colon motility were increased in most SERT -/- animals; however, the increase in motility (diarrhea) occasionally alternated irregularly with decreased motility (constipation). The watery diarrhea is probably attributable to the potentiation of serotonergic signaling in SERT -/- mice, whereas the transient constipation may be caused by episodes of enhanced 5-HT release leading to 5-HT receptor desensitization. PMID- 11487659 TI - Amphetamine-induced plasticity of AMPA receptors in the ventral tegmental area: effects on extracellular levels of dopamine and glutamate in freely moving rats. AB - Previous electrophysiological studies suggested that the initiation of behavioral sensitization to cocaine and amphetamine involves a transient increase in AMPA receptor responsiveness in the ventral tegmental area (VTA). To test this, we used in vivo microdialysis to examine the effects of intra-VTA administration of AMPA (10 microm) and NMDA (100 microm) on dopamine (DA) and glutamate efflux in the VTA and the nucleus accumbens (NAC), an important target of VTA DA neurons. We compared rats treated for 5 d with saline or 5 mg/kg amphetamine and withdrawn for 3 or 10-14 d. After 3 d of withdrawal, intra-VTA AMPA increased both NAC and VTA DA levels to a greater extent in the amphetamine group, whereas NMDA produced similar effects in the saline and amphetamine groups. This enhanced responsiveness to AMPA was no longer evident in rats tested 10-14 d after the last injection. In addition, intra-VTA AMPA but not NMDA increased both VTA and NAC glutamate levels in rats tested 3 d after the last injection of amphetamine but not in saline controls. After 10-14 d, the responsiveness of glutamate levels to AMPA was no longer evident in the NAC but persisted in the VTA. Additional studies indicated that the glutamate effect in the NAC may involve increased responsiveness of DA receptors within the NAC. These findings establish an in vivo animal model with which to explore the consequences of repeated drug administration for AMPA receptor plasticity in the VTA. They also indicate that repeated amphetamine leads to potentiated interactions between DA and glutamate transmission. PMID- 11487660 TI - Dopamine D1 and NMDA receptors mediate potentiation of basolateral amygdala evoked firing of nucleus accumbens neurons. AB - Interactions between the basolateral amygdala (BLA) and the nucleus accumbens (NAc) mediate reward-related processes that are modulated by mesoaccumbens dopamine (DA) transmission. The present in vivo electrophysiological study assessed: (1) changes in the firing probability of submaximal BLA-evoked single neuronal firing activity in the NAc after tetanic stimulation of the BLA, and (2) the functional roles of DA and NMDA receptors in these processes. Tetanic stimulation of the BLA potentiated BLA-evoked firing activity of NAc neurons for a short duration ( approximately 25 min). This short-term potentiation was associated with an increase in DA oxidation currents that was monitored with chronoamperometry. Systemic or iontophoretic application before BLA tetanus of the D(1) receptor antagonist SCH23390, but not the D(2) receptor antagonist sulpiride, abolished the potentiation of BLA-evoked NAc activity, whereas administration of SCH23390 3 min after tetanus had no effect. However, systemic administration of the NMDA antagonist 3-(2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid (CPP), either before or after BLA tetanus, abolished the potentiation of BLA-evoked firing of NAc neurons. These data suggest that higher frequency activity in BLA efferents can autoregulate their excitatory influence over neural activity of NAc neurons by facilitating the release of DA and activating both DA D(1) and NMDA receptors. This may represent a cellular mechanism that facilitates approach behaviors directed toward reward-related stimuli that are mediated by BLA-NAc circuitries. PMID- 11487661 TI - Physiological correlates of comodulation masking release in the mammalian ventral cochlear nucleus. AB - Comodulation masking release (CMR) enhances the detection of signals embedded in wideband, amplitude-modulated maskers. At least part of the CMR is attributable to across-frequency processing, however, the relative contribution of different stages in the auditory system to across-frequency processing is unknown. We have measured the responses of single units from one of the earliest stages in the ascending auditory pathway, the ventral cochlear nucleus, where across frequency processing may take place. A sinusoidally amplitude-modulated tone at the best frequency of each unit was used as a masker. A pure tone signal was added in the dips of the masker modulation (reference condition). Flanking components (FCs) were then added at frequencies remote from the unit best frequency. The FCs were pure tones amplitude modulated either in phase (comodulated) or out of phase (codeviant) with the on-frequency component. Psychophysically, this CMR paradigm reduces within-channel cues while producing an advantage of approximately 10 dB for the comodulated condition in comparison with the reference condition. Some of the recorded units showed responses consistent with perceptual CMR. The addition of the comodulated FCs produced a strong reduction in the response to the masker modulation, making the signal more salient in the poststimulus time histograms. A decision statistic based on d' showed that threshold was reached at lower signal levels for the comodulated condition than for reference or codeviant conditions. The neurons that exhibited such a behavior were mainly transient chopper or primary-like units. The results obtained from a subpopulation of transient chopper units are consistent with a possible circuit in the cochlear nucleus consisting of a wideband inhibitor contacting a narrowband cell. A computational model was used to confirm the feasibility of such a circuit. PMID- 11487662 TI - Group I metabotropic glutamate receptors elicit epileptiform discharges in the hippocampus through PLCbeta1 signaling. AB - Activation of metabotropic glutamate receptors (mGluRs) produces multiple effects in cortical neurons, resulting in the emergence of network activities including epileptiform discharges. The cellular mechanisms underlying such network responses are largely unknown. We examined the properties of group I mGluR mediated cellular responses in CA3 neurons and attempted to determine their role in the generation of the network activities. Group I mGluR stimulation causes depolarization of hippocampal neurons. This depolarization is primarily mediated by two sets of conductance change: the opening of a voltage-dependent cationic conductance (mediating I(mGluR(V))) and the closing of a voltage-independent (background) K(+) conductance. I(mGluR(V)) was no longer elicited by group I mGluR agonists in the presence of U73122, a phospholipase C (PLC) blocker. Also, the current could not be activated in hippocampal CA3 neurons from PLCbeta1 knock out mice. In contrast, suppression of PLC signaling did not affect the group I mGluR-mediated suppression of background K(+) conductance. Thus, the suppression of the background K(+) conductance occurred upstream to PLC activation, whereas the generation of I(mGluR(V)) occurred downstream to PLC activation. Group I mGluR agonists normally elicited rhythmic single cell and population burst responses in the CA3 neurons. In the absence of an I(mGluR(V)) response, CA3 neurons in slices prepared from PLCbeta1-/- mutant mice could no longer generate these responses. The results suggest that I(mGluR(V)) expression in CA3 hippocampal neuron is PLCbeta1-dependent and that I(mGluR(V)) plays a necessary role in the generation of rhythmic single cell bursts and synchronized epileptiform discharges in the CA3 region of the hippocampus. PMID- 11487663 TI - Experience- and age-related outgrowth of intrinsic neurons in the mushroom bodies of the adult worker honeybee. AB - A worker honeybee performs tasks within the hive for approximately the first 3 weeks of adult life. After this time, it becomes a forager, flying repeatedly to collect food outside of the hive for the remainder of its 5-6 week life. Previous studies have shown that foragers have an increased volume of neuropil associated with the mushroom bodies, a brain region involved in learning, memory, and sensory integration. We report here that growth of the mushroom body neuropil in adult bees occurs throughout adult life and continues after bees begin to forage. Studies using Golgi impregnation asked whether the growth of the collar region of the mushroom body neuropil was a result of growth of the dendritic processes of the mushroom body intrinsic neurons, the Kenyon cells. Branching and length of dendrites in the collar region of the calyces were strongly correlated with worker age, but when age-matched bees were directly compared, those with foraging experience had longer, more branched dendrites than bees that had foraged less or not at all. The density of Kenyon cell dendritic spines remained constant regardless of age or behavioral state. Older and more experienced foragers therefore have a greater total number of dendritic spines in the mushroom body neuropil. Our findings indicate that, under natural conditions, the cytoarchitectural complexity of neurons in the mushroom bodies of adult honeybees increases as a function of increasing age, but that foraging experience promotes additional dendritic branching and growth. PMID- 11487664 TI - Action spectrum for melatonin regulation in humans: evidence for a novel circadian photoreceptor. AB - The photopigment in the human eye that transduces light for circadian and neuroendocrine regulation, is unknown. The aim of this study was to establish an action spectrum for light-induced melatonin suppression that could help elucidate the ocular photoreceptor system for regulating the human pineal gland. Subjects (37 females, 35 males, mean age of 24.5 +/- 0.3 years) were healthy and had normal color vision. Full-field, monochromatic light exposures took place between 2:00 and 3:30 A.M. while subjects' pupils were dilated. Blood samples collected before and after light exposures were quantified for melatonin. Each subject was tested with at least seven different irradiances of one wavelength with a minimum of 1 week between each nighttime exposure. Nighttime melatonin suppression tests (n = 627) were completed with wavelengths from 420 to 600 nm. The data were fit to eight univariant, sigmoidal fluence-response curves (R(2) = 0.81-0.95). The action spectrum constructed from these data fit an opsin template (R(2) = 0.91), which identifies 446-477 nm as the most potent wavelength region providing circadian input for regulating melatonin secretion. The results suggest that, in humans, a single photopigment may be primarily responsible for melatonin suppression, and its peak absorbance appears to be distinct from that of rod and cone cell photopigments for vision. The data also suggest that this new photopigment is retinaldehyde based. These findings suggest that there is a novel opsin photopigment in the human eye that mediates circadian photoreception. PMID- 11487665 TI - The contribution of activity-dependent synaptic plasticity to classical conditioning in Aplysia. AB - Plasticity at central synapses has long been thought to be the most likely mechanism for learning and memory, but testing that idea experimentally has proven to be difficult. For this reason, we have developed a simplified preparation of the Aplysia siphon withdrawal reflex that allows one to examine behavioral learning and memory while simultaneously monitoring synaptic connections between individual identified neurons in the CNS. We previously found that monosynaptic connections from LE siphon sensory neurons to LFS siphon motor neurons make a substantial contribution to the reflex in the siphon withdrawal preparation (Antonov et al., 1999a). We have now used that preparation to assess the contribution of various cellular mechanisms to classical conditioning of the reflex with a siphon tap conditioned stimulus (CS) and tail shock unconditioned stimulus (US). We find that, compared with unpaired training, paired training with the CS and US produces greater enhancement of siphon withdrawal and evoked firing of LFS neurons, greater facilitation of the complex PSP elicited in an LFS neuron by the siphon tap, and greater facilitation of the monosynaptic PSP elicited by stimulation of a single LE neuron. Moreover, the enhanced facilitation of monosynaptic LE-LFS PSPs is greater for LE neurons that fire during the siphon tap and correlates significantly with the enhancement of siphon withdrawal and evoked firing of the LFS neurons. These results provide the most direct evidence to date that activity-dependent plasticity at specific central synapses contributes to behavioral conditioning and support the idea that synaptic plasticity is a mechanism of learning and memory more generally. PMID- 11487666 TI - Alleviation of a selective age-related relational memory deficit in mice by pharmacologically induced normalization of brain retinoid signaling. AB - Vitamin A and its derivatives, the retinoids, have been implicated recently in the synaptic plasticity of the hippocampus and might therefore play a role in associated cognitive functions. Acting via transcription factors, retinoids can regulate gene expression via their nuclear receptors [retinoic acid receptors (RARs) and retinoid X receptors]. In a series of experiments, the present study investigated the possible role of age-related downregulation of retinoid-mediated transcription events in the cognitive decline seen in aged mice. We observed that the brain (and hippocampal) levels of retinoid receptors and the expression of specific associated target genes were restored to presenescent (adult) levels in aged mice after acute administration (150 microg/kg, s.c.) of retinoic acid (RA). These effects of RA, however, could be abolished by the coadministration of an RAR antagonist. RA was also demonstrated to alleviate the age-related deficit in the CA1 long-term potentiation efficacy of aged mice in vivo. Moreover, RA was found to alleviate completely the performance deficit of aged mice to the control level in a two-stage spatial discrimination paradigm designed to assess relational memory. This promnesic effect of RA was again susceptible to abolition by RAR antagonist treatment. The parallel molecular, cellular, and behavioral correlates associated with the decrease of retinoid receptor expression and its normalization demonstrated here suggest that the fine regulation of retinoid mediated gene expression is fundamentally important to optimal brain functioning and higher cognition. Specifically, a naturally occurring dysregulation of retinoid-mediated molecular events might be a potential etiological factor for cognitive deterioration during senescence. PMID- 11487667 TI - Cortical slow oscillatory activity is reflected in the membrane potential and spike trains of striatal neurons in rats with chronic nigrostriatal lesions. AB - Neurons in the basal ganglia output nuclei display rhythmic burst firing after chronic nigrostriatal lesions. The thalamocortical network is a strong endogenous generator of oscillatory activity, and the striatum receives a massive projection from the cerebral cortex. Actually, the membrane potential of striatal projection neurons displays periodic shifts between a very negative resting potential (down state) and depolarizing plateaus (up states) during which they can fire action potentials. We hypothesized that an increased excitability of striatal neurons may allow transmission of cortical slow rhythms through the striatum to the remaining basal ganglia in experimental parkinsonism. In vivo intracellular recordings revealed that striatal projection neurons from rats with chronic nigrostriatal lesions had a more depolarized membrane potential during both the down and up states and an increased firing probability during the up events. Furthermore, lesioned rats had significantly fewer silent neurons than control rats. Simultaneous recordings of the frontal electrocorticogram and membrane potential of striatal projection neurons revealed that the signals were oscillating synchronously in the frequency range 0.4-2 Hz, both in control rats and rats with chronic nigrostriatal lesions. Spreading of the slow cortical rhythm is limited by the very low firing probability of control rat neurons, but a slow oscillation is well reflected in spike trains of approximately 60% of lesioned rat neurons. These findings provide in vivo evidence for a role of dopamine in controlling the flow of cortical activity through the striatum and may be of outstanding relevance for understanding the pathophysiology of Parkinson's disease. PMID- 11487668 TI - Chattering and differential signal processing in identified motion-sensitive neurons of parallel visual pathways in the chick tectum. AB - At least three identified cell types in the stratum griseum centrale (SGC) of the chick optic tectum mediate separate pathways from the retina to different subdivisions of the thalamic nucleus rotundus. Two of these, SGC type I and type II, constitute the major direct inputs to rotundal subdivisions that process various aspects of visual information, e.g., motion and luminance changes. Here, we examined the responses of these cell types to somatic current injection and synaptic input. We used a brain slice preparation of the chick tectum and applied whole-cell patch recordings, restricted electrical stimulation of dendritic endings, and subsequent labeling with biocytin. Type I neurons responded with regular sequences of bursts ("chattering") to depolarizing current injection. Electrical stimulation of retinal afferents evoked a sharp-onset EPSP/burst response that was blocked with CNQX. The sharp-onset EPSP/burst response to synaptic stimulation persisted when the soma was hyperpolarized, thus suggesting the presence of dendritic spike generation. In contrast, the type II neurons responded to depolarizing current injection solely with an irregular sequence of individual spikes. Electrical stimulation of retinal afferents led to slow and long-lasting EPSPs that gave rise to one or several action potentials. In conclusion, the morphological distinct SGC type I and II neurons also have different response properties to retinal inputs. This difference is likely to have functional significance for the differential processing of visual information in the separate pathways from the retina to different subdivisions of the thalamic nucleus rotundus. PMID- 11487669 TI - Compensation for the effects of head acceleration on jaw movement in speech. AB - Recent studies have demonstrated the ability of subjects to adjust the control of limb movements to counteract the effects of self-generated loads. The degree to which subjects change control signals to compensate for these loads is a reflection of the extent to which forces affecting movement are represented in motion planning. Here, we have used empirical and modeling studies to examine whether the nervous system compensates for loads acting on the jaw during speech production. As subjects walk, loads to the jaw vary with the direction and magnitude of head acceleration. We investigated the patterns of jaw motion resulting from these loads both in locomotion alone and when locomotion was combined with speech production. In locomotion alone, jaw movements were shown to vary systematically in direction and magnitude in relation to the acceleration of the head. In contrast, when locomotion was combined with speech, variation in jaw position during both consonant and vowel production was substantially reduced. Overall, we have demonstrated that the magnitude of load associated with head acceleration during locomotion is sufficient to produce a systematic change in the position of the jaw. The absence of variation in jaw position during locomotion with speech is thus consistent with the idea that in speech, the control of jaw motion is adjusted in a predictive manner to offset the effects of head acceleration. PMID- 11487670 TI - The use of cyanoacrylate adhesives in peripheral embolization. AB - Although liquid adhesives or glue have been used as embolic agents for nearly three decades, experience with them outside of neurointerventional indications is generally limited. Cyanoacrylates are the main liquid adhesives used in the vascular system and have an important role in managing vascular abnormalities, especially arteriovenous malformations. Vascular occlusion results as these agents polymerize on exposure to the ions in blood. A description of the properties, biologic interactions, techniques of use, and indications for acrylic embolization in the peripheral circulation is especially pertinent at this time because of the recent approval of n-butyl cyanoacrylate by the United States Food and Drug Administration. PMID- 11487671 TI - Intravascular radiation therapy after balloon angioplasty of narrowed femoropopliteal arteries to prevent restenosis: results of the PARIS feasibility clinical trial. AB - PURPOSE: To conduct a feasibility study to assess the feasibility, safety, and outcome of endoluminal gamma radiation therapy after balloon angioplasty of superficial femoral artery (SFA) lesions. MATERIALS AND METHODS: Forty patients with claudication were enrolled in the study and underwent percutaneous transluminal angioplasty (PTA) of SFA lesions with a mean lesion length of 9.8 cm +/- 3.0 and a mean reference vessel diameter of 5.2 mm +/- 3.1. After successful PTA, a segmented centering balloon catheter was positioned to cover the PTA site. The patients were then transported to the radiation oncology suite and treated with a microSelectron HDR afterloader with use of an Ir-192 source with a prescribed dose of 14 Gy, 2 mm into the vessel wall. Ankle-brachial index (ABI) and Rutherford score were evaluated at 1, 6, and 12 months after the procedure and angiographic follow-up was conducted at 6 months. RESULTS: Radiation was delivered successfully to 35 of 40 patients. There were no procedural complications. Exercise and rest ABI were higher at 1 year (0.72 +/- 0.26 and 0.89 +/- 0.18, respectively) compared to baseline (0.51 +/- 0.25 and 0.67 +/- 0.17, respectively). Maximum walking time on a treadmill increased from 3.41 min +/- 2.41 to 4.43 min +/- 2.49 at 30 days and was 4.04 min +/- 2.8 at 12 months. The angiographic binary restenosis rate at 6 months was 17.2% and the clinical restenosis rate at 12 months was 13.3%. There were no angiographic or clinical adverse events related to the radiation therapy. CONCLUSIONS: Intraarterial radiation after PTA of SFA lesions with use of high-dose rate gamma radiation is feasible and safe. The angiographic and clinical improvements are sustainable at 1 year and represent a potent antirestenotic therapy for the treatment of narrowed peripheral arteries. PMID- 11487672 TI - Predictors of long-term patency after femoropopliteal angioplasty: results from the STAR registry. AB - PURPOSE: To identify variables predictive of long-term patency after femoropopliteal angioplasty. MATERIALS AND METHODS: The primary patency of 219 limbs in 205 patients from a multicenter registry who underwent femoropopliteal angioplasty between January 1, 1992, and December 31, 1994, was prospectively monitored with a combination of angiography, noninvasive hemodynamic testing, and clinical outcome. Patient demographic, angiographic, and hemodynamic variables were examined alone and in combination to determine effect on long-term primary patency. Each limb was graded as Category 1-4 according to the American Heart Association (AHA) criteria for arterial lesions, and differences in outcome for each category were examined. Primary patency and intergroup analysis were determined with use of the Kaplan-Meier method and log-rank test, respectively. Cox proportional hazards models were used to calculate relative risks for predictive variables. RESULTS: Primary patency rates for all limbs (on an intent to-treat basis) at 12, 24, and 36 months were 87% +/- 3%, 80% +/- 3%, and 69% +/- 5%, respectively. Primary patency at 48 and 60 months was 55% +/- 7%. Poor tibial runoff (single tibial vessel with 50%-99% stenosis or occlusion) was most predictive of occlusion (relative risk 8.5, P <.0001). The presence of diabetes or renal failure was associated with lower long-term patency (relative risk 5.5 and 4.0, P <.0001 and.0002, respectively). Long-term patency was higher with AHA Category 1 lesions (P =.006), and no significant difference in patency was observed between Category 2 and 3 lesions (P =.65). A multivariate Cox proportional hazards model showed only the stratified runoff score and the presence of diabetes to be significant determinants of long-term patency. CONCLUSION: Poor tibial runoff is most predictive of lower long-term patency rates. Diabetes is also independently associated with lower long-term patency rates. The criteria that distinguish Category 2 and 3 lesions do not predict differences in long-term patency, nor do they serve to identify lesions best treated with surgical bypass. This suggests that indications for femoral angioplasty can be extended to include longer and more complex Category 3 lesions. PMID- 11487673 TI - Randomized study to compare PTA alone versus PTA with Palmaz stent placement for femoropopliteal lesions. AB - PURPOSE: The purpose of this study was to evaluate whether percutaneous transluminal angioplasty (PTA) combined with Palmaz stent placement provides long term advantages compared to PTA alone after 34 months of follow-up in the femoropopliteal region. MATERIALS AND METHODS: Thirty patients randomized to undergo PTA in combination with stent placement and 23 patients randomized to undergo PTA alone were evaluated. RESULTS: Mean follow-up (+/-SD) for the PTA group was 33.8 months (+/- 8.7) and for the Palmaz group 29.1 months (+/- 6.2), with a maximum follow-up period of 39 months for both groups. No significant differences in primary or secondary patency rates could be observed at 12 or 39 months. After 39 months, the primary patency rate for PTA alone was 68.4% and the secondary patency rate was 89.5%; the primary patency rate for PTA with stent placement was 62% and the secondary patency rate was 90%. CONCLUSIONS: The results of this study show that even after a long-term follow up of more than 3 years, PTA with stent placement in the femoropopliteal artery does not produce better results than PTA alone, although it does provide better initial luminal gain after the procedure. PMID- 11487674 TI - Placement of Hemobahn stent-grafts in femoropopliteal arteries: early experience and midterm results in 18 patients. AB - PURPOSE: To determine the outcome of a new polytetrafluoroethylene (PTFE)-covered stent-graft in the treatment of occlusive lesions of the femoropopliteal arteries. MATERIALS AND METHODS: Eighteen patients with intermittent claudication (Fontaine stages IIb-IV; Rutherford classes I/2-III/5) and occlusive lesions of the superficial femoral and/or popliteal artery were treated with use of a new PTFE-covered stent-graft (Hemobahn). Indication for stent-graft insertion was an unsatisfactory result after balloon dilation. RESULTS: Successful stent-graft placement was achieved in 17 patients (94%). In one patient, additional insertion of a Palmaz stent was necessary because of a stent-graft wrinkle. Mean ankle brachial index increased from 0.72 +/- 0.15 preoperatively to 0.94 +/- 0.17 postoperatively (P =.028). Fifteen patients (83%) experienced an initial improvement of at least one clinical stage. Primary patency was 61% +/- 11% at 3 months and 49% +/- 12% at 6 months. Stent-graft occlusion was observed in 13 patients (72%). In four patients, the stent-graft was reopened percutaneously by balloon angioplasty (n = 2) and/or rotational thrombectomy (n = 2). Therefore, the secondary patency rate was 67% +/- 11% at 3 months and 61% +/- 11% at 6 months. Patency rates proved to be significantly lower for stent-grafts longer than 10 cm (P =.03). Intimal hyperplasia at the proximal or distal end of the stent-graft was observed in seven patients (39%). Complications such as access site hematoma (n = 3), distal thromboembolism (n = 2), and abscess formation around the stent-graft (n = 1) were observed. CONCLUSION: Despite excellent initial angiographic results, the patency rate in lesions treated with the Hemobahn stent-graft in this small study was unsatisfactory. Use of stent-grafts longer than 10 cm and the occurrence of intimal hyperplasia proved to be major factors in the reduction of long-term patency rates. PMID- 11487675 TI - Recombinant tissue plasminogen activator (alteplase) for restoration of flow in occluded central venous access devices: a double-blind placebo-controlled trial- the Cardiovascular Thrombolytic to Open Occluded Lines (COOL) efficacy trial. AB - PURPOSE: Central venous access devices (CVADs) are a mainstay of current medical therapy but often become occluded by thrombus. Tissue plasminogen activator (alteplase), at a dose of 2 mg per 2 mL, has been shown to be effective in restoring flow to catheters proven by radiographic contrast injection to be occluded by thrombus. The purpose of this double-blind placebo-controlled multicenter trial was to determine the efficacy of alteplase in occluded catheters without earlier contrast injections or radiographic examinations. MATERIALS AND METHODS: Patients were eligible for inclusion if blood could not be withdrawn from their catheter after a period of normal function of at least 48 hours. Single or multiple catheters, peripherally inserted central catheters, catheters with valves, and implanted ports were eligible; catheters used for hemodialysis were not included. Patients were randomly assigned to one of two groups. In one group, patients received a first dose of 2 mg alteplase followed, if needed, by a second dose of 2 mg alteplase and a third dose of placebo. The other group received placebo first followed by one 2-mg dose of alteplase and then a second, if needed. Each dose was allowed to dwell for 2 hours and ability to withdraw blood from the catheter was reassessed. The endpoint was restoration of the ability to withdraw and infuse through the catheter. One hundred forty nine patients were randomized: 74 received placebo first, 75 received alteplase first. RESULTS: After the first 2-hour treatment, function was restored to 74% in the alteplase arm and 17% in the placebo arm (P <.0001 compared to placebo). After one or two treatments, function was restored in 90% of patients. There were no serious study-drug-related adverse events, no intracranial hemorrhage, no major hemorrhage, and no embolic events. CONCLUSION: Infusion of alteplase appeared to be safe and effective in restoring flow to occluded catheters without need for pretreatment radiographic evaluation. PMID- 11487676 TI - Right gastric artery embolization to prevent acute gastric mucosal lesions in patients undergoing repeat hepatic arterial infusion chemotherapy. AB - PURPOSE: The purpose of the study was to investigate the technical outcome and clinical effect of right gastric artery (RGA) embolization to prevent acute gastric mucosal lesions caused by influx of anticancer agents into the RGA in patients undergoing repeat hepatic arterial infusion chemotherapy (HAIC). MATERIALS AND METHODS: In 217 patients with malignant hepatic tumors, we attempted RGA embolization with use of metallic coils and/or a mixture of n-butyl cyanoacrylate (n-BCA) and iodized oil, along with the embolization of the gastroduodenal artery. After this procedure, an infusion catheter was placed radiologically and HAIC was performed. We then evaluated the technical outcome and clinical effect of RGA embolization. RESULTS: RGA embolization was technically successful in 201 of 217 patients (93%). Major complications--nausea, epigastric pain, and fever--were noted in 12%, 4%, and 2% of successful cases, respectively, and were treated conservatively. Recanalization occurred in 4% (nine of 201) of the patients. Eventually, sufficient RGA embolization was achieved in 192 patients. The incidence of acute gastric mucosal lesions confirmed endoscopically was only 3% (five of 192) in patients with sufficient RGA embolization, whereas it was 36% (nine of 25) in patients without sufficient RGA embolization, with a significant difference (P <.01). CONCLUSION: RGA embolization is a highly feasible procedure that can reduce the incidence of acute gastric mucosal lesions associated with HAIC. PMID- 11487677 TI - Risk factors for liver abscess formation after hepatic chemoembolization. AB - PURPOSE: To assess the frequency and risk factors for liver abscess after hepatic chemoembolization. MATERIALS AND METHODS: The authors performed retrospective analysis of 397 chemoembolization procedures in 157 patients. All patients received prophylactic intravenous antibiotics before the procedure and 5 days of oral antibiotics after the procedure. The association between abscess formation and risk factors was determined with use of chi(2) analysis and the Fisher exact test and expressed as an odds ratio. RESULTS: Liver abscess occurred in seven of 157 patients (4.5%) after eight of 397 procedures (2.0%) at a mean of 19 d +/- 7 after chemoembolization. No patients had neutropenia. Organisms isolated reflected intestinal flora. Six patients required percutaneous drainage for 35 d +/- 29. The seventh patient required drainage for the remainder of life as a result of a nonhealing biliary fistula. Three of 24 patients with neuroendocrine tumors had abscesses (12.5%; odds ratio, 4.6; 95% CI, 0.96-22.1; P =.07), as did three of 14 patients with gastrointestinal sarcomas (21%; odds ratio, 9.5; 95% CI, 1.9-47.8; P =.016), and one of two with pancreatic adenocarcinoma. Six of the seven patients with abscesses underwent a Whipple procedure before chemoembolization. Only one patient with a history of a Whipple procedure did not develop an hepatic abscess. In the absence of a bilioenteric anastomosis, abscess occurred in only one of 150 patients (0.7%), or one of 383 procedures (0.3%). The odds ratio for liver abscess among patients with a bilioenteric anastomosis was 894 (95% CI, 50-16,000; P <.0001). CONCLUSION: Earlier bilioenteric anastomosis is the major determinant of liver abscess formation after hepatic chemoembolization. The prophylaxis regimen used failed to prevent abscess formation in patients with earlier bilioenteric anastomosis. PMID- 11487678 TI - The porcine hemodialysis access model. AB - PURPOSE: To create a porcine hemodialysis access model that reliably reproduces intimal hyperplasia (IH) of the outflow vein similar to that which causes access failure in human patients undergoing dialysis treatments. MATERIALS AND METHODS: Surgical technique for creation of side-to-side iliac-artery-to-ipsilateral-iliac vein (IAV) native fistulas and IAV conduits was optimized in three standard-bred pigs. Persistent patency of fistulas and conduits was demonstrated in two additional pigs allowed to survive for 1 week. IAV fistulas and contralateral 2 cm polytetrafluoroethylene IAV conduits were created in five additional pigs. Venous outflow from these fistulas and conduits was evaluated with venography and intravascular ultrasound (IVUS) immediately after creation (day 0) and at 2-week intervals for as long as 64 days. Animals were killed at 30 days (n = 1), 42 days (n = 2), or 64 days (n = 2), and the arteries, veins, and conduits were evaluated histologically. RESULTS: IAV native fistulas remained patent until the animals' death and conduits remained patent for at least 14 days in four of five pigs; both the fistula and conduit likely occluded before 16-day follow-up in the fifth pig. At 42-64 days, venography demonstrated maximum fistula outflow vein diameter stenoses of 53%-76% and maximum conduit outflow vein stenoses of 44%-84%, and IVUS demonstrated maximum area stenoses of 64%-86% and 43%-82%, respectively. Three of five conduits occluded, one before 16-day follow-up, one between 14 and 28 days, and the other after 42 days. Histologic sections demonstrated IH predominantly affecting the veins at the anastomoses and central (cephalad) to the anastomoses in all pigs. CONCLUSION: This porcine model reproduces IH in the fistula or conduit outflow vein with measurable stenosis. Such a model might allow relevant preclinical evaluation of interventional devices and techniques intended to reduce the effects of IH in human patients undergoing dialysis treatments. PMID- 11487679 TI - Use of ethylene vinyl alcohol copolymer for tubal sterilization by selective catheterization in rabbits. AB - PURPOSE: To assess the efficacy of ethylene vinyl alcohol copolymer (Uryx) in nonsurgically occluding the fallopian tube and achieving tubal sterilization in the rabbit model. MATERIALS AND METHODS: Ten mature virgin female New England rabbits underwent transvaginal selective bilateral fallopian tube cannulation with use of a coaxial catheter system under general anesthesia. Selective salpingography was performed bilaterally to assess patency of the fallopian tubes. Ethylene vinyl alcohol copolymer was injected unilaterally through a microcatheter to completely fill the middle portion of the tube. Three to seven days after injection, each animal was bred. Conception was determined by ultrasonography (US) 7-19 days after effective breeding. If pregnant, the rabbit was killed. Otherwise, it was permitted to rebreed until pregnancy was achieved. Histologic specimens of the fallopian tubes were prepared and analyzed. RESULTS: Patency of the fallopian tubes was demonstrated bilaterally in all animals by the free spillage of contrast material into the peritoneum. The delivery of ethylene vinyl alcohol copolymer into the fallopian tubes was successful in all animals but one, in which most of the plug almost immediately extruded into the uterus. Pregnancy was detected by US in the untreated fallopian tube in the nine rabbits that were receptive to breeding. No pregnancies were detected in the injected side. Histologic analysis demonstrated variable degrees of occlusion, fibrosis, and inflammation, with the majority of specimens demonstrating mild to moderate inflammation and moderate to marked fibrosis. CONCLUSION: Ethylene vinyl alcohol copolymer can reliably be placed nonsurgically via the transvaginal approach into the fallopian tubes with use of a coaxial catheter system. Ethylene vinyl alcohol copolymer appears to result in less fibrosis than previously investigated agents and demonstrates a 100% early sterilization rate in the rabbit model. PMID- 11487680 TI - Accuracy of CO(2) angiography in vessel diameter assessment: a comparative study of CO(2) versus iodinated contrast material in a porcine model. AB - PURPOSE: To compare, with use of intravascular ultrasound (IVUS) as an internal reference standard in a porcine model, arterial diameters measured from arteriograms obtained with use of CO(2) to those obtained with use of iodinated contrast material (ICM). MATERIALS AND METHODS: In nine pigs, digital subtraction angiograms (DSAs) were obtained in the aorta and iliac arteries to compare vessel diameters measured with use of CO(2) to those measured with use of ICM. These measurements were divided by measurements made with use of intravascular ultrasound (IVUS) to yield a DSA/IVUS ratio. Differences between ICM and CO(2) were compared with analysis of variance to assess the effect of location (aorta vs iliac), contrast material used (ICM vs CO(2)), and position (posteroanterior, right anterior oblique, or left anterior oblique). Secondary analysis compared measurements of dependent and nondependent iliac arteries and compared the use of hand-injected CO(2) to that of CO(2) injected by an injector. RESULTS: The DSA/IVUS ratio was 70.7% +/- 4.4% with ICM use and 69.6% +/- 6.3% with CO(2) use, which did not represent a significant difference (P =.311). Animal position had no effect (P =.477). Underestimation was worse in the iliac arteries than in the aorta (67.4% +/- 1.5% vs 71.4% +/- 1.7%; P =.038). There was no difference in nondependent (P =.163) arteries, but CO(2) underestimated dependent iliac artery size more than ICM did (66.3% +/- 4.8% vs 70.3% +/- 5.4%; P =.051). Vessel diameter was underestimated more with the CO(2) injector than with hand-injected CO(2) (64.3% +/- 2.3% vs 71.7% +/- 1.7%; P <.0001). CONCLUSION: There is no difference in diameter underestimation between CO(2) and ICM in this animal model. Hand-injection of CO(2) causes less underestimation of vessel diameter than does the CO(2) injector. PMID- 11487681 TI - Use of a TrapEase device as a temporary caval filter. AB - Inferior vena cava (IVC) thrombosis in younger patients presents a difficult management problem and is associated with a significant incidence of pulmonary embolism (PE). Treatment options include anticoagulation, mechanical thrombectomy, or thrombolytic therapy, often in combination with placement of a filter above the thrombus. The authors report the use of a permanent filter in a temporary fashion while performing thrombectomy and thrombolysis of an IVC thrombus. PMID- 11487682 TI - Anchoring a migrating inferior vena cava stent with use of a T-fastener. AB - An attempt to treat symptomatic stenosis of the inferior vena cava in a patient with metastatic liver disease was complicated by migration of a Wallstent into the right atrium. Effective palliation was achieved by insertion of a second stent, which was anchored by transhepatic insertion of a T-fastener into the intracaval stent. This anchoring maneuver was performed safely under sonographic and fluoroscopic guidance. PMID- 11487683 TI - A new twist on the Waltman loop for uterine fibroid embolization. AB - The usual ipsilateral approach for uterine artery embolization is to form a Waltman loop. Newer nonbraided 4-F catheters can be problematic because of their tendency to kink or unfold while the loop is being formed. Herein, a modification of the Cope suture technique is described by which a Waltman loop is formed with use of a 4-0 Tevdek suture that allows the catheter to be folded back on itself, drawing it into the ipsilateral iliac artery. If necessary, the catheter can be prevented from unlooping by twisting the catheter on itself to lock it in position. The technique allows for efficient catheterization of the ipsilateral uterine artery. PMID- 11487684 TI - Transient arterial hypotension induced by polymethylmethacrylate injection during percutaneous vertebroplasty. PMID- 11487685 TI - A sense of self: the role of DNA sequence elimination in allopolyploidization. PMID- 11487686 TI - Plant photobiology 2001: a thousand points of enlightenment from receptor structures to ecological adaptation. PMID- 11487687 TI - The RNA world in plants: post-transcriptional control III. PMID- 11487689 TI - Allopolyploidy-induced rapid genome evolution in the wheat (Aegilops-Triticum) group. AB - To better understand genetic events that accompany allopolyploid formation, we studied the rate and time of elimination of eight DNA sequences in F1 hybrids and newly formed allopolyploids of Aegilops and TRITICUM: In total, 35 interspecific and intergeneric F1 hybrids and 22 derived allopolyploids were analyzed and compared with their direct parental plants. The studied sequences exist in all the diploid species of the Triticeae but occur in only one genome, either in one homologous pair (chromosome-specific sequences [CSSs]) or in several pairs of the same genome (genome-specific sequences [GSSs]), in the polyploid wheats. It was found that rapid elimination of CSSs and GSSs is a general phenomenon in newly synthesized allopolyploids. Elimination of GSSs was already initiated in F1 plants and was completed in the second or third allopolyploid generation, whereas elimination of CSSs started in the first allopolyploid generation and was completed in the second or third generation. Sequence elimination started earlier in allopolyploids whose genome constitution was analogous to natural polyploids compared with allopolyploids that do not occur in nature. Elimination is a nonrandom and reproducible event whose direction was determined by the genomic combination of the hybrid or the allopolyploid. It was not affected by the genotype of the parental plants, by their cytoplasm, or by the ploidy level, and it did not result from intergenomic recombination. Allopolyploidy-induced sequence elimination occurred in a sizable fraction of the genome and in sequences that were apparently noncoding. This finding suggests a role in augmenting the differentiation of homoeologous chromosomes at the polyploid level, thereby providing the physical basis for the diploid-like meiotic behavior of newly formed allopolyploids. In our view, this rapid genome adjustment may have contributed to the successful establishment of newly formed allopolyploids as new species. PMID- 11487688 TI - KNAT2: evidence for a link between knotted-like genes and carpel development. AB - The KNAT2 (for KNOTTED-like from Arabidopsis thaliana 2) homeobox gene is expressed in the vegetative apical meristem. It is also active during flower development, suggesting a function in the structuring of flowers. To investigate its role, we used a DEXAMETHASONE (DEX)-inducible system to generate transgenic plants that overexpressed a fusion of KNAT2 with the hormone binding domain of the glucocorticoid receptor. DEX-induced plants were similar to plants overexpressing the closely related KNAT1 gene, indicating overlapping functions, although we observed differences as well. In particular, KNAT2-GR activation induced ectopic carpel features. First, KNAT2 induced the homeotic conversion of nucellus into carpel-like structures. Second, KNAT2 induced stigmatic papillae on rosette leaves in the ap2-5 background. Third, ectopic expression of the carpel identity gene AGAMOUS (AG) was observed in carpels and ovules. Interestingly, the homeotic conversion was not dependent on AG activity, because it was maintained in the ag-1 ap2-5 double mutant. Therefore, our data indicate that KNAT2 also must activate other carpel regulators. Together, these results suggest that KNAT2 plays a role in carpel development. PMID- 11487690 TI - Sequence elimination and cytosine methylation are rapid and reproducible responses of the genome to wide hybridization and allopolyploidy in wheat. AB - Interspecific or intergeneric hybridization, followed by chromosome doubling, can lead to the formation of new allopolyploid species. Recent studies indicate that allopolyploid formation is associated with genetic and epigenetic changes, although little is known about the type of changes that occur, how rapidly they occur, and the type of sequences involved. To address these matters, we have surveyed F1 hybrids between diploid species from the wheat (Aegilops and Triticum) group and their derived allotetraploids by screening a large number of loci using amplified fragment length polymorphism and DNA gel blot analysis and by assaying the extent of cytosine methylation. We found that sequence elimination is one of the major and immediate responses of the wheat genome to wide hybridization or allopolyploidy, that it affects a large fraction of the genome, and that it is reproducible. In one cross between AE: sharonensis x AE: umbellulata, 14% of the loci from AE: sharonensis were eliminated compared with only 0.5% from AE: umbellulata, with most changes occurring in the F1 hybrid. In contrast, crosses between AE: longissima x T. urartu showed that sequence elimination was more frequent after chromosome doubling. Alterations in cytosine methylation occurred in approximately 13% of the loci, either in the F1 hybrid or in the allopolyploid. For eight of nine bands that were isolated, the sequences that underwent elimination corresponded to low-copy DNA, whereas alterations in methylation patterns affected both repetitive DNA sequences, such as retrotransposons, and low-copy DNA in approximately equal proportions. PMID- 11487691 TI - The Arabidopsis dual-affinity nitrate transporter gene AtNRT1.1 (CHL1) is activated and functions in nascent organ development during vegetative and reproductive growth. AB - The AtNRT1.1 (CHL1) transporter provides a primary mechanism for nitrate uptake in Arabidopsis and is expected to localize to the epidermis and cortex of the mature root, where the bulk of nitrate uptake occurs. Using fusions to GFP/GUS marker genes, we found CHL1 expression concentrated in the tips of primary and lateral roots, with very low signals in the epidermis and cortex. A time-course study showed that CHL1 is activated in the primary root tip early in seedling development and at the earliest stages of lateral root formation. Strong CHL1 expression also was found in shoots, concentrated in young leaves and developing flower buds but not in the shoot meristem. These expression patterns were confirmed by immunolocalization and led us to examine CHL1 function specifically in the growth of developing organs. chl1 mutants showed a reduction in the growth of nascent roots, stems, leaves, and flower buds. The growth of nascent primary roots was inhibited in the mutants even in the absence of added nitrate, whereas elongation of lateral root primordia was inhibited specifically at low nitrate and acidic pH. Interestingly, chl1 mutants also displayed a late-flowering phenotype. These results indicate that CHL1 is activated and functions in the growth of nascent organs in both shoots and roots during vegetative and reproductive growth. PMID- 11487692 TI - ORE9, an F-box protein that regulates leaf senescence in Arabidopsis. AB - Senescence is a sequence of biochemical and physiological events that constitute the final stage of development. The identification of genes that alter senescence has practical value and is helpful in revealing pathways that influence senescence. However, the genetic mechanisms of senescence are largely unknown. The leaf of the oresara9 (ore9) mutant of Arabidopsis exhibits increased longevity during age-dependent natural senescence by delaying the onset of various senescence symptoms. It also displays delayed senescence symptoms during hormone-modulated senescence. Map-based cloning of ORE9 identified a 693-amino acid polypeptide containing an F-box motif and 18 leucine-rich repeats. The F-box motif of ORE9 interacts with ASK1 (Arabidopsis Skp1-like 1), a component of the plant SCF complex. These results suggest that ORE9 functions to limit leaf longevity by removing, through ubiquitin-dependent proteolysis, target proteins that are required to delay the leaf senescence program in Arabidopsis. PMID- 11487693 TI - Expression of Arabidopsis GAI in transgenic rice represses multiple gibberellin responses. AB - Bioactive gibberellins (GAs) are essential endogenous regulators of plant growth. GA signaling is mediated via GAI, a nuclear member of the GRAS family of plant transcription factors. Previous experiments have suggested that GAI is a GA derepressible repressor of plant growth. Here we test this hypothesis by examining the effects of the expression of Arabidopsis GAI in transgenic Basmati rice. High-level expression of GAI caused dwarfism and reduced GA responses, and the strength of this effect was correlated with the level of transgene expression. In particular, the expression of GAI abolished the GA-mediated induction of rice aleurone alpha-amylase activity, thus implicating GAI orthologs in the well-characterized cereal aleurone GA response. The GA derepressible repressor model predicts that high-level expression of GAI should confer dwarfism, and these observations are consistent with this prediction. PMID- 11487694 TI - The PET1-CMS mitochondrial mutation in sunflower is associated with premature programmed cell death and cytochrome c release. AB - In mammals, mitochondria have been shown to play a key intermediary role in apoptosis, a morphologically distinct form of programmed cell death (PCD), for example, through the release of cytochrome c, which activates a proteolytic enzyme cascade, resulting in specific nuclear DNA degradation and cell death. In plants, PCD is a feature of normal development, including the penultimate stage of anther development, leading to dehiscence and pollen release. However, there is little evidence that plant mitochondria are involved in PCD. In a wide range of plant species, anther and/or pollen development is disrupted in a class of mutants termed CMS (for cytoplasmic male sterility), which is associated with mutations in the mitochondrial genome. On the basis of the manifestation of a number of morphological and biochemical markers of apoptosis, we have shown that the PET1-CMS cytoplasm in sunflower causes premature PCD of the tapetal cells, which then extends to other anther tissues. These features included cell condensation, oligonucleosomal cleavage of nuclear DNA, separation of chromatin into delineated masses, and initial persistence of mitochondria. In addition, immunocytochemical analysis revealed that cytochrome c was released partially from the mitochondria into the cytosol of tapetal cells before the gross morphological changes associated with PCD. The decrease in cytochrome c content in mitochondria isolated from male sterile florets preceded a decrease in the integrity of the outer mitochondrial membrane and respiratory control ratio. Our data suggest that plant mitochondria, like mammalian mitochondria, play a key role in the induction of PCD. The tissue-specific nature of the CMS phenotype is discussed with regard to cellular respiratory demand and PCD during normal anther development. PMID- 11487695 TI - Developmental abnormalities and reduced fruit softening in tomato plants expressing an antisense Rab11 GTPase gene. AB - A cDNA clone from tomato fruit encodes a protein with strong homology with the rab11/YPT3 class of small GTPases that is thought to be involved in the control of protein trafficking within cells. The gene, LeRab11a, showed a pattern consistent with a single copy in DNA gel blots. The corresponding mRNA was developmentally regulated during fruit ripening, and its expression was inhibited in several ripening mutants. Its reduced expression in the Never-ripe mutant indicates that it may be induced by ethylene in fruit. The ripening-induced expression in tissues that are undergoing cell wall loosening immediately suggests a possible role in trafficking of cell wall-modifying enzymes. The message also was produced in leaves and flowers but not in roots. Antisense transformation was used to generate a "mutant phenotype." Antisense fruit changed color as expected but failed to soften normally. This was accompanied by reduced levels of two cell wall hydrolases, pectinesterase and polygalacturonase. There were other phenotypic effects in the plants, including determinate growth, reduced apical dominance, branched inflorescences, abnormal floral structure, and ectopic shoots on the leaves. In some plants, ethylene production was reduced. These data suggest an alternative or additional role in exocytosis or endocytosis of homeotic proteins, hormone carriers, or receptors. PMID- 11487696 TI - Ethylene inhibits the Nod factor signal transduction pathway of Medicago truncatula. AB - Legumes form a mutualistic symbiosis with bacteria collectively referred to as rhizobia. The bacteria induce the formation of nodules on the roots of the appropriate host plant, and this process requires the bacterial signaling molecule Nod factor. Although the interaction is beneficial to the plant, the number of nodules is tightly regulated. The gaseous plant hormone ethylene has been shown to be involved in the regulation of nodule number. The mechanism of the ethylene inhibition on nodulation is unclear, and the position at which ethylene acts in this complex developmental process is unknown. Here, we used direct and indirect ethylene application and inhibition of ethylene biosynthesis, together with comparison of wild-type plants and an ethylene-insensitive supernodulating mutant, to assess the effect of ethylene at multiple stages of this interaction in the model legume Medicago truncatula. We show that ethylene inhibited all of the early plant responses tested, including the initiation of calcium spiking. This finding suggests that ethylene acts upstream or at the point of calcium spiking in the Nod factor signal transduction pathway, either directly or through feedback from ethylene effects on downstream events. Furthermore, ethylene appears to regulate the frequency of calcium spiking, suggesting that it can modulate both the degree and the nature of Nod factor pathway activation. PMID- 11487697 TI - Somatic and germinal excision activities of the Arabidopsis transposon Tag1 are controlled by distinct regulatory sequences within Tag1. AB - Various sequences within Tag1, the endogenous transposon of Arabidopsis, were examined to determine how Tag1 excision and expression are regulated. The 5' intron for the major 2.3-kb Tag1 transcript was found to be critical for the accumulation of Tag1 transcripts and for high rates of somatic excision. This was true for the autonomous element in cauliflower mosaic virus 35S-Tag1-beta glucuronidase constructs and for a two-component system using the 35S promoter to produce Tag1 transposase and a beta-glucuronidase::dTag1 marker construct to score for excision. The 3' introns of Tag1, although not needed for high transposase expression in primary transgenic plants, were important for maintaining high levels of somatic excision and accumulation of the major but not the minor Tag1 transcripts in subsequent generations. With both 5' and 3' introns present, exchanging the 5' promoter region of Tag1 with the 35S promoter did not affect the timing of Tag1 excision significantly, but it did disrupt germinal excision. Removal of the 5' intron did not abolish germinal excision activity, however. These results indicate that somatic and germinal excision of Tag1 are differentially controlled, with the 5' promoter region being critical for germinal excision activity and the 5' intron playing an important role for somatic excision, possibly via intron-mediated enhancement. PMID- 11487698 TI - EMF1, a novel protein involved in the control of shoot architecture and flowering in Arabidopsis. AB - Shoot architecture and flowering time in angiosperms depend on the balanced expression of a large number of flowering time and flower meristem identity genes. Loss-of-function mutations in the Arabidopsis EMBRYONIC FLOWER (EMF) genes cause Arabidopsis to eliminate rosette shoot growth and transform the apical meristem from indeterminate to determinate growth by producing a single terminal flower on all nodes. We have identified the EMF1 gene by positional cloning. The deduced polypeptide has no homology with any protein of known function except a putative protein in the rice genome with which EMF1 shares common motifs that include nuclear localization signals, P-loop, and LXXLL elements. Alteration of EMF1 expression in transgenic plants caused progressive changes in flowering time, shoot determinacy, and inflorescence architecture. EMF1 and its related sequence may belong to a new class of proteins that function as transcriptional regulators of phase transition during shoot development. PMID- 11487699 TI - Activation of salicylic acid-induced protein kinase, a mitogen-activated protein kinase, induces multiple defense responses in tobacco. AB - The activation of mitogen-activated protein kinases (MAPKs) is one of the earliest responses in plants challenged by avirulent pathogens or cells treated with pathogen-derived elicitors. Expression of a constitutively active MAPK kinase, NtMEK2(DD), in tobacco induces the expression of defense genes and hypersensitive response-like cell death, which are preceded by the activation of two endogenous MAPKs, salicylic acid-induced protein kinase (SIPK) and wounding induced protein kinase (WIPK). However, the roles that SIPK and WIPK each play in the process are unknown. Here we report that SIPK alone is sufficient to activate these defense responses. In tobacco leaves transiently transformed with SIPK under the control of a steroid-inducible promoter, the induction of SIPK expression after the application of dexamethasone, a steroid, leads to an increase of SIPK activity. The increase of SIPK activity is dependent on the phosphorylation of newly synthesized SIPK by its endogenous upstream kinase. In contrast, the expression of WIPK under the same conditions fails to increase its activity, even though the protein accumulates to a similar level. Studies using chimeras of SIPK and WIPK demonstrated that the C terminus of SIPK contains the molecular determinant for its activation, which is rather surprising because the N termini of SIPK and WIPK are more divergent. SIPK has been implicated previously in the regulation of both plant defense gene activation and hypersensitive response-like cell death based on evidence from pharmacological studies using kinase inhibitors. This gain-of-function study provided more direct evidence for its role in the signaling of multiple defense responses in tobacco. PMID- 11487700 TI - G2/M-phase-specific transcription during the plant cell cycle is mediated by c Myb-like transcription factors. AB - Plant B-type cyclin genes are expressed specifically in late G2- and M-phases during the cell cycle. Their promoters contain a common cis-acting element, called the MSA (M-specific activator) element, that is necessary and sufficient for periodic promoter activation. This motif also is present in the tobacco kinesin-like protein gene NACK1, which is expressed with timing similar to that of B-type cyclin genes. In this study, we show that G2/M-phase-specific activation of the NACK1 promoter also is regulated by the MSA element, suggesting that a defined set of G2/M-phase-specific genes are coregulated by an MSA mediated mechanism. In a search for MSA binding factors by yeast one-hybrid screening, we identified three different Myb-like proteins that interact specifically with the MSA sequence. Unlike the majority of plant Myb-like proteins, these Myb proteins, NtmybA1, NtmybA2, and NtmybB, have three imperfect repeats in the DNA binding domain, as in animal c-Myb proteins. During the cell cycle, the level of NtmybB mRNA did not change significantly, whereas the levels of NtmybA1 and A2 mRNAs fluctuated and peaked at M-phase, when B-type cyclin genes were maximally induced. In transient expression assays, NtmybA1 and A2 activated the MSA-containing promoters, whereas NtmybB repressed them. Furthermore, expression of NtmybB repressed the transcriptional activation mediated by NtmybA2. Our data show that a group of plant Myb proteins that are structurally similar to animal c-Myb proteins have unexpected roles in G2/M-phase by modulating the expression of B-type cyclin genes and may regulate a suite of coexpressed genes. PMID- 11487701 TI - The Arabidopsis sex1 mutant is defective in the R1 protein, a general regulator of starch degradation in plants, and not in the chloroplast hexose transporter. AB - Starch is the major storage carbohydrate in higher plants and of considerable importance for the human diet and for numerous technical applications. In addition, starch can be accumulated transiently in chloroplasts as a temporary deposit of carbohydrates during ongoing photosynthesis. This transitory starch has to be mobilized during the subsequent dark period. Mutants defective in starch mobilization are characterized by high starch contents in leaves after prolonged periods of darkness and therefore are termed starch excess (sex) mutants. Here we describe the molecular characterization of the Arabidopsis sex1 mutant that has been proposed to be defective in the export of glucose resulting from hydrolytic starch breakdown. The mutated gene in sex1 was cloned using a map based cloning approach. By complementation of the mutant, immunological analysis, and analysis of starch phosphorylation, we show that sex1 is defective in the Arabidopsis homolog of the R1 protein and not in the hexose transporter. We propose that the SEX1 protein (R1) functions as an overall regulator of starch mobilization by controlling the phosphate content of starch. PMID- 11487702 TI - Maize chromomethylase Zea methyltransferase2 is required for CpNpG methylation. AB - A cytosine DNA methyltransferase containing a chromodomain, Zea methyltransferase2 (Zmet2), was cloned from maize. The sequence of ZMET2 is similar to that of the Arabidopsis chromomethylases CMT1 and CMT3, with C terminal motifs characteristic of eukaryotic and prokaryotic DNA methyltransferases. We used a reverse genetics approach to determine the function of the Zmet2 gene. Plants homozygous for a Mutator transposable element insertion into motif IX had a 13% reduction in methylated cytosines. DNA gel blot analysis of these plants with methylation-sensitive restriction enzymes and bisulfite sequencing of a 180-bp knob sequence showed reduced methylation only at CpNpG sites. No reductions in methylation were observed at CpG or asymmetric sites in heterozygous or homozygous mutant plants. Our research shows that chromomethylase Zmet2 is required for in vivo methylation of CpNpG sequences. PMID- 11487703 TI - Dynamic recruitment of Cdc2 to specific microtubule structures during mitosis. AB - A-type cyclin-dependent kinases (CDKs), also known as cdc2, are central to the orderly progression of the cell cycle. We made a functional Green Fluorescent Protein (GFP) fusion with CDK-A (Cdc2-GFP) and followed its subcellular localization during the cell cycle in tobacco cells. During interphase, the Cdc2 GFP fusion protein was found in both the cytoplasm and the nucleus, where it was highly resistant to extraction. In premitotic cells, a bright and narrow equatorial band appeared on the cell surface, resembling the late preprophase band, which disintegrated within 10 min as followed by time-lapse images. Cdc2 GFP was not found on prophase spindles but left the chromatin soon after this stage and associated progressively with the metaphase spindle in a microtubule dependent manner. Arresting cells in mitosis through the stabilization of microtubules by taxol further enhanced the spindle-localized pool of Cdc2-GFP. Toward the end of mitosis, Cdc2-GFP was found at the midzone of the anaphase spindle and phragmoplast; eventually, it became focused at the midline of these microtubule structures. In detergent-extracted cells, the Cdc2-GFP remained associated with mitotic structures. Retention on spindles was prevented by pretreatment with the CDK-specific inhibitor roscovitine and was enhanced by the protein phosphatase inhibitor okadaic acid. Furthermore, we demonstrate that both the endogenous CDK-A and Cdc2-GFP were cosedimented with taxol-stabilized plant microtubules from cell extracts and that Cdc2 activity was detected together with a fraction of polymerized tubulin. These data provide evidence that the A-type CDKs associate physically with mitotic structures in a microtubule-dependent manner and may be involved in regulating the behavior of specific microtubule arrays throughout mitosis. PMID- 11487704 TI - Peroxisomal metabolic function is required for appressorium-mediated plant infection by Colletotrichum lagenarium. AB - Peroxisomes are organelles that perform a wide range of metabolic functions in eukaryotic cells. However, their role in fungal pathogenesis is poorly understood. Here we report that ClaPEX6, an ortholog of PEX6, is required for the fungus Colletotrichum lagenarium to infect host plants. ClaPEX6 was identified in random insertional mutagenesis experiments aimed at elucidating genes involved in pathogenesis. Functional analysis, using a green fluorescent protein cassette containing the peroxisomal targeting signal1 (PTS1), revealed that import of PTS1 containing proteins is impaired in clapex6 mutants generated by targeted gene disruption. Failure of growth on fatty acids shows an inability of fatty acid beta-oxidation in these mutants. These results indicate that disruption of ClaPEX6 impairs peroxisomal metabolism, even though clapex6 mutants show normal growth and conidiation in nutrient-rich conditions. The clapex6 mutants formed small appressoria with severely reduced melanization that failed to form infectious hyphae. These data indicate that peroxisomes are necessary for appressorium-mediated penetration of host plants. The addition of glucose increased the pathogenicity of clapex6 mutants, suggesting that the glucose metabolic pathway can compensate partially for peroxisomes in phytopathogenicity. PMID- 11487705 TI - Repression domains of class II ERF transcriptional repressors share an essential motif for active repression. AB - We reported previously that three ERF transcription factors, tobacco ERF3 (NtERF3) and Arabidopsis AtERF3 and AtERF4, which are categorized as class II ERFs, are active repressors of transcription. To clarify the roles of these repressors in transcriptional regulation in plants, we attempted to identify the functional domains of the ERF repressor that mediates the repression of transcription. Analysis of the results of a series of deletions revealed that the C-terminal 35 amino acids of NtERF3 are sufficient to confer the capacity for repression of transcription on a heterologous DNA binding domain. This repression domain suppressed the intermolecular activities of other transcriptional activators. In addition, fusion of this repression domain to the VP16 activation domain completely inhibited the transactivation function of VP16. Comparison of amino acid sequences of class II ERF repressors revealed the conservation of the sequence motif (L)/(F)DLN(L)/(F)(x)P. This motif was essential for repression because mutations within the motif eliminated the capacity for repression. We designated this motif the ERF-associated amphiphilic repression (EAR) motif, and we identified this motif in a number of zinc-finger proteins from wheat, Arabidopsis, and petunia plants. These zinc finger proteins functioned as repressors, and their repression domains were identified as regions that contained an EAR motif. PMID- 11487706 TI - A decrease in soleus muscle force generation in rats after downhill running. AB - The purpose of the present study was to investigate the immediate and 48-hr post exercise effects of eccentric contraction-biased exercise on the contractile properties of the soleus muscle in situ. Adult male Wistar rats were categorised into sedentary control rats (n = 10), rats studied immediately (n = 10), and rats studied 48 hours after the exercise (n = 10). The exercise protocol consisted of a 90-min intermittent downhill running (-16 degrees, 16 m/min) on a motor-driven treadmill. The contractile properties of the soleus muscle were recorded following i.p. chloral hydrate anaesthesia. Isometric twitch force (Pt), time-to peak tension (TPT), half-relaxation time (1/2 RT), and tetanic force at stimulation frequencies of 40, 80, and 100 Hz were recorded. A low-frequency muscle fatigue protocol (stimulation at 4 Hz for 5 min) was applied to test for fatigability. The main findings indicated that Pt generation dropped both immediately and 48 hr after the exercise, while tetanic force was partially restored after 48 hr. Exercise-induced E-C coupling failure and contractile machinery disorganisation due to muscle injury are put forward as the main force reduction causes. PMID- 11487707 TI - The incidence of hematuria in middle distance track running. AB - The purpose of this study was to establish if middle distance track athletes experience hematuria during their competitive season interval workouts and, if so, what type of workout based on intensity and distance was most associated with hematuria. During a 4-week observational period, athletes (n = 10) underwent reagent strip urinalysis before and after their twice weekly interval sessions. Positive samples for hematuria were analyzed microscopically to accurately determine red blood cell (RBC) loss. Seventy-one individual interval workouts were observed, of which 32 cases of hematuria were reported. Nine cases of hematuria exhibited >100 RBC per High Power Field (Hpf). Furthermore, 90% of the athletes experienced post-workout hematuria at least once. The highest incidence of hematuria was observed after workouts run at 110% of VO(2peak) over short (600 1,500 m) to moderate (1,501-3,000 m) distances. All post-exercise cases of hematuria resolved within 2 hr of recovery. PMID- 11487708 TI - The VO(2) response at the onset of severe intensity exercise. AB - Fourteen participants achieved a peak VO(2) of 2573 +/- 738 ml a min-1 in approximately 3 1/2-min cycle ergometer tests. However, in the first 45 s of exercise, formal description of the phase II of the V(O)2 response indicated that the V(O)2 was projecting toward a value of 3451 +/- 1599 ml x min(-1), well above the peak VO(2)peak and not different from the predicted O(2) demand of 3389 +/- 800 ml x min(-1). We conclude that, at the onset of exercise in the severe intensity domain, VO(2) is initially driven toward the O(2) demand, and then is limited by the achievable VO(2) (VO(2peak)). PMID- 11487709 TI - Respiratory muscle injury: evidence to date and potential mechanisms. AB - Respiratory muscle dysfunction associated with ventilatory loading may be partially attributed to respiratory muscle injury. Exertion-induced muscle injury can be defined as structural alterations of the muscle, however, a better understanding of the biochemical, morphologic, and functional correlates of injured respiratory muscles will facilitate discrimination of how injury, fatigue, and weakness contribute to respiratory muscle dysfunction. In addition to the increased loads associated with lung disease, many factors such as poor arterial blood gases, immobilization, sepsis, decreased nutrition, and corticosteroids may increase susceptibility to exertion-induced respiratory muscle injury. Respiratory muscle injury in humans is not well-described, however, more extensive evidence has been shown in animal models of increased ventilatory loading. Potential mechanisms of respiratory muscle injury are mechanical stress, metabolic stress, and inflammation. In order to optimize therapeutic interventions, a better understanding of these mechanisms and the patients that are most susceptible to respiratory muscle injury needs to be determined. PMID- 11487710 TI - Physical activity assessment options within the context of the Canadian Physical Activity, Fitness, and Lifestyle Appraisal. AB - The Canadian Physical Activity, Fitness, and Lifestyle Appraisal (CPAFLA) is a standardized battery of tests that is commonly used to assess asymptomatic individuals. This paper reviews evidence concerning the reliability, validity, and utility of the procedure currently used to assess habitual physical activity in the CPAFLA and surveys available physical activity assessment options. Special considerations related to age, gender, ethnicity, seasonal variations, and within week variance are discussed. The potential assessment options are discussed with consideration given to the logistical and pragmatic constraints inherent in the CPAFLA. New technologies for future assessment of physical activity are briefly presented and recommendations for future research forwarded. PMID- 11487711 TI - Duration of therapy in metastatic breast cancer: management using Herceptin. AB - Despite progressive developments in therapeutic interventions, including surgery, radiotherapy and chemotherapy, there has been no major improvement in the survival of women with metastatic breast cancer (MBC). Based on knowledge of tumor growth patterns, approaches addressing this issue have included increasing chemotherapy dose or dose density and extending the duration of therapy. However, only the latter approach has resulted in improved clinical benefit, although not survival; and its use is restricted by the cumulative toxicity of chemotherapeutic agents. Therefore, the best hope for improved survival lies with new classes of anticancer drug and particularly biological agents. This review focuses on the first oncogene-targeted therapy for human epidermal growth factor receptor-2 (HER2)+ MBC patients. The humanized anti-HER2 monoclonal antibody Herceptin has proven clinical benefits in HER2+ MBC patients, most importantly improved survival, and is rapidly becoming a standard of care for these patients. In contrast to the fixed number of cycles used for chemotherapeutic agents, Herceptin is administered until disease progression, with some data suggesting that continuation beyond disease progression should be investigated. The preclinical and clinical findings on which the current recommended duration of Herceptin therapy are based are reviewed and alternative strategies are discussed. PMID- 11487712 TI - Thymidylate synthase as a molecular target for drug discovery using the National Cancer Institute's Anticancer Drug Screen. AB - Thymidylate synthase (TS) is a critical cellular target for cancer chemotherapeutics, particularly the fluoropyrimidine and antifolate classes of antineoplastic agents. One of the primary mechanisms of clinical insensitivity to these agents is through the overexpression of the target enzyme, TS. Thus, there is a need for the development of agents which selectively target TS overexpressing malignant cells. To this end, we conducted a search for agents which potentially selectively target TS-overexpressing cells using two separate algorithms for identifying such compounds in the NCI Drug Repository by comparing cytotoxicity profiles of 30000 compounds with the TS expression levels measured by Western blot analysis in 53 cell lines. Using the traditional COMPARE analysis we were unable to identify compounds which maintain a selective ability to kill high TS-expressing cells in a subsequent four cell line validation assay. A new algorithm, termed COMPARE Effect Clusters analysis, enabled the identification of a particular drug cluster which contained compounds that maintained a selective ability to kill TS-overexpressing cell lines in the validation assay. While the identified compounds were selectively cytotoxic to TS-overexpressing cells, we found that they were not specifically targeting TS as a mechanism of action. Apparently, the overexpression of TS was providing a marker for sensitivity. This identified class of compounds which appears to be selectively cytotoxic against cells which overexpress TS may be useful for the development of therapeutics for those whose cancers overexpress TS de novo. PMID- 11487713 TI - Chemical and enzymatic stability of a cyclic depsipeptide, the novel, marine derived, anti-cancer agent kahalalide F. AB - Kahalalide F is a cyclic depsipeptide isolated from the Hawaiian mollusk Elysia rufescens. This compound is under present phase I clinical investigations as an anti-tumor drug. The role of possible metabolic reactions of this drug in (pre )clinical investigations has not yet been explored. The first results for kahalalide F in this field of research are given in this paper. The chemical degradation of kahalalide F was investigated under acid, neutral and alkaline conditions using high-performance liquid chromatography with ultraviolet detection. The half-lives at 80 degrees C were 1.1, 20 and 8.6 h at pH 0, 1 and 7, respectively. At 26 degrees C and pH 11, the half-life was 1.65 h. At pH 7 and 11, only one reaction product of kahalalide F was observed, kahalalide G, the hydrolyzed lactone product of kahalalide F. At pH 0 and 1, additional reaction products emerged. Metabolic conversion of kahalalide F was tested in vitro using three different enzyme systems based on pooled human microsomes, pooled human plasma and uridine 5'-diphosphoglucuronyl transferase, respectively. The incubated samples were analyzed using the same chromatographic technique as for the degradation samples. Biotransformations were not observed under these conditions and, therefore, it is concluded that kahalalide F is a metabolically stable drug. PMID- 11487714 TI - Population pharmacokinetics and dynamics in phase II studies of the novel bioreductive alkylating cytotoxic indoloquinone EO9. AB - Population pharmacokinetic-dynamic analysis was prospectively integrated in the clinical phase II programme of EO9 to determine the population pharmacokinetic profile in a larger population of patients, to estimate individual patient pharmacokinetic parameters, and to investigate relationships between drug exposure and clinical outcome. A sparse sampling method was developed, which involved three sampling times, and was implemented during course 1. A Bayesian algorithm was used to estimate individual pharmacokinetic parameters, in particular total plasma clearance (CL) of EO9 and area under the curve (AUC). In total, samples were collected of 85 (65%) of the patients. Pharmacokinetic evaluation was successful in 61 (72%) of the sampled patients. CL of EO9 showed substantial variability (median 5.08 l/min; range 2.67-6.42) and was of the same magnitude as in the phase I study where full pharmacokinetic profiles were used. No significant relationships were noticed between exposure parameters and safety, but overall limited toxicity was observed. No tumor responses were documented. Prospective implementation of large-scale population pharmacokinetic-dynamic analysis is feasible and may generate important findings, in particular when tumor responses and relevant toxicity are observed. PMID- 11487715 TI - Pharmacokinetics of oral cyclosporin A when co-administered to enhance the oral absorption of paclitaxel. AB - The objective of this study was to evaluate the pharmacokinetics of oral cyclosporin A (CsA) when co-administered to enhance the oral absorption of paclitaxel. Patients received oral paclitaxel in doses of 60-360 mg/m(2) in combination with a dose of oral CsA of 15 mg/kg. Dose escalation of paclitaxel from 60 to 300 mg/m(2) resulted in a significant decrease in the area under the concentration-time curve (AUC) of CsA from 24.4+/-9.9 to 17.6+/-2.8 mg/l.h (p=0.03) (n=28). In conclusion, increases in the paclitaxel dose resulted in a decrease in the AUC of CsA. This observation may be explained by the increase in the co-solvent Cremophor EL of paclitaxel causing reduced absorption of CsA. PMID- 11487716 TI - Sequence-dependent cytotoxicity of combination chemotherapy using paclitaxel, carboplatin and bleomycin in human lung and ovarian cancer. AB - Combination chemotherapy for non-small cell lung cancer (NSCLC) and ovarian cancer typically consists of a regimen of a taxane such as paclitaxel and a platinum-containing agent. Bleomycin, which halts cell cycle progression at G2 phase, is an agent which might thereby increase taxane cytotoxicity. The goal of this study was to evaluate the effect of different paclitaxel-platinum or paclitaxel-bleomycin schedules on cytotoxicity in human NSCLC and ovarian cancer cells. The simultaneous combination of paclitaxel and carboplatin exhibited simple additivity in vitro, while sequential exposure studies indicated that carboplatin followed by paclitaxel produced greater than additive cytotoxicity using the isobologram analysis of combinatorial effects. In contrast, the simultaneous combination of paclitaxel and bleomycin consistently exhibited greater than additive effects indicating a potentially synergistic combination. Sequential exposure studies of bleomycin followed by paclitaxel produced similar synergistic findings. Experiments in SCID mice evaluating the combinations of paclitaxel and bleomycin supported the in vitro results, as significantly enhanced A549 lung tumor growth inhibition was observed when paclitaxel was administered 1 h after bleomycin. The synergistic activity shown by the combination of bleomycin and paclitaxel indicates a potentially beneficial novel combination for treatment of NSCLC and ovarian cancer. PMID- 11487717 TI - Synthesis and antiproliferative activity of [2-(phthaloylamino)alkyl]triphenyl phosphonium derivatives against K562 cell line. AB - Given the reported cytotoxicity of phthaloylaminoethyltriphenylphosphonium bromide 2a in the P-388 cell line, we have developed new [2 (phthaloylamino)alkyl]phosphonium derivatives 2b-e and evaluated their cytotoxic activity. These compounds have been synthetized from N,N-phthaloylaminoalcohols and triphenylphosphonium hydrobromide via a one-pot reaction. 2a was found inactive in the K562 cell line, but 2c-e exhibited a cytotoxic activity with IC50 values about 1 microM. PMID- 11487718 TI - Subcellular redistribution of BAX during apoptosis induced by anticancer drugs. AB - BAX is the 192-amino acid, 21-kDa protein which is ubiquitously distributed in normal tissues and is regarded as a tumor suppressor sensitizing malignant cells to anticancer drugs. In spite of many studies, the molecular mechanism of BAX action is still obscure. In the present study subcellular BAX translocations in human colon adenocarcinoma COLO 205 cells exposed to various anticancer drugs [camptothecin (CPT), etoposide (ETO), staurosporine (STP), 2-chloro-2' deoxyadenosine (2CdA) and nimesulide (NIM)] was examined. Cells were grown on coverslips under optimal conditions (10% FCS/DMEM) or were stimulated to apoptosis with the drugs examined. Laser scanning cytometry was applied for the quantitative analysis of BAX expression, and distribution in the cytoplasmic (BAX Cf) and nuclear (BAX Nf) area. BAX maximal pixel (BAX MP), the parameter corresponding to aggregation of BAX in the cell, was also measured. All examined drugs increased the number of cells with high BAX MP, reaching the peak at 60 min after drug administration. The most pronounced effect was in the case of 2CdA, CPT and STP. The increase in BAX MP was observed only when antibody recognizing the 43-61 amino acid sequence was used. When antibody binding the N-terminal epitope (11-30 amino acid sequence) was applied, the number of cells expressing high BAX MP significantly decreased. These results indicate that apoptotic stimuli delivered by anticancer drugs led to aggregation of BAX in cancer cells, which is dependent on BAX activation by its cleavage at the N-terminal epitope and exposure of the BH3 domain. It was shown that BAX Nf increased in cells treated with CPT, STP, ETO, 2CdA and NIM, whereas BAX Cf rose after STP and NIM. The increase in BAX Nf and, occurring in most treatments, the increase in the BAX Nf:Cf ratio indicates a BAX shift from the cytoplasm to the nucleus. Furthermore, staining with different antibodies showed that only the activated form of BAX was translocated to the nucleus. Immunoelectron microscopy revealed that CPT-induced apoptosis was associated with translocation of BAX from the cytosol to organellar membranes (mitochondrial, Golgi apparatus and endoplasmic reticulum) and via nuclear envelope pores to the nucleus, occurring within 60-180 min of cell exposure to the drug. The subcellular translocations of BAX preceded in time the appearance of morphological symptoms of apoptosis. In conclusion, (i) in spite of different molecular mechanisms of apoptosis induction by the anticancer drugs examined, BAX remains a common link in the chain of reactions leading to cell death, and (ii) BAX activation and subcellular translocations from the cytosol to organellar membranes and nucleus are key cellular responses to drugs bearing proapoptotic properties. PMID- 11487719 TI - Toxicity of irinotecan (CPT-11) and hepato-renal dysfunction. AB - Various clinical and laboratory parameters have been investigated for their ability to predict toxicity arising from the use of the anticancer drug, irinotecan (CPT-11). In particular, patients deficient in the conjugation of SN 38, a metabolite of CPT-11, are known to be at greater risk. We describe one case of a patient with metastatic colorectal cancer treated with a single dose of CPT 11 at 125 mg/m(2). Although this patient lacked any known predictive factors for toxicity, he experienced severe side-effects several days later. We hypothesized that the toxicity in this patient was due to compromised SN-38 conjugation. Plasma samples were analyzed by reversed-phase high-performance liquid chromatography assay for CPT-11 and its metabolites at 96, 144, 168, 192 and 288 h post-administration. We observed that the concentrations of both the parent drug and its metabolites were markedly raised (11- to 60-fold expected). Additionally the estimated terminal half-lives were 1.5-7 times those expected (29.5, 101, 39.6 and 41.8 h for CPT-11, APC, SN-38G and SN-38, respectively). We conclude that the toxicity in this patient was not caused by deficient SN-38 conjugation, but by decreased drug excretion through both hepatic and renal routes. PMID- 11487720 TI - A case of glassy cell carcinoma of the uterine cervix effectively responding to chemotherapy with paclitaxel and carboplatin. AB - Glassy cell carcinoma (GCC) of the uterine cervix is a highly malignant tumor and has a poor prognosis. As yet, no effective systemic chemotherapy to this tumor has been reported. Here we describe a case of recurrent GCC that responded to paclitaxel and carboplatin combination treatment. The patient, a 32-year-old woman, with clinical staging FIGO IB1 disease had a radical hysterectomy and postoperative radiotherapy. Three months after initial treatment, she had a relapse as peritoneal dissemination, which was confirmed in the second surgery (adnectomy) and which did not respond to platinum-based conventional chemotherapy (cisplatin, adriamycin, cyclophosphamide and carboplatin, etoposide). The recurrent peritoneal tumors responded well to paclitaxel and carboplatin combination treatment. An elevated serum concentration of carcinoembryonic antigen (672 ng/ml) was reduced to 14.4 ng/ml by six such courses. Peritoneal histopathology confirmed a complete response in the third surgery (ileostomy) for adhesive ileus by the radiotherapy. This is the first report of effective systemic chemotherapy with paclitaxel and carboplatin to recurrent GCC of the uterine cervix. PMID- 11487721 TI - A rare case of advanced ovarian carcinoma who developed difficulty walking 25 days after treatment with weekly paclitaxel. AB - Although taxol has shown significant activity in advanced ovarian cancer, peripheral neuropathy is likely to become the major dose-limiting toxicity. We describe a case of advanced ovarian carcinoma who developed difficulty walking because of marked pain in the lower extremities and loss of proprioception 25 days after treatment with weekly taxol (80 mg/m(2)x3). PMID- 11487724 TI - Protein kinase C and cerebral vasospasm. AB - Twenty-five years after the discovery of protein kinase C (PKC), the physiologic function of PKC, and especially its role in pathologic conditions, remains a subject of great interest with 30,000 studies published on these aspects. In the cerebral circulation, PKC plays a role in the regulation of myogenic tone by sensitization of myofilaments to calcium. Protein kinase C phosphorylates various ion channels including augmenting voltage-dependent Ca2+ channels and inhibiting K+ channels, which both lead to vessel contraction. These actions of PKC amplify vascular reactivity to different agonists and may be critical in the regulation of cerebral artery tone during vasospasm. Evidence accumulated during at least the last decade suggest that activation of PKC in cerebral vasospasm results in a delayed but prolonged contraction of major arteries after subarachnoid hemorrhage. Most of the experimental results in vitro or in animal models support the view that PKC is involved in cerebral vasospasm. Implication of PKC in cerebral vasospasm helps explain increased arterial narrowing at the signal transduction level and alters current perceptions that the pathophysiology is caused by a combination of multiple receptor activation, hemoglobin toxicity, and damaged neurogenic control. Activation of protein kinase C also interacts with other signaling pathways such as myosin light chain kinase, nitric oxide, intracellular Ca2+, protein tyrosine kinase, and its substrates such as mitogen activated protein kinase. Even though identifying PKC revolutionized the understanding of cerebral vasospasm, clinical advances are hampered by the lack of clinical trials using selective PKC inhibitors. PMID- 11487722 TI - Cardenolides and cancer. PMID- 11487725 TI - The key role of caveolin-1 in estrogen-mediated regulation of endothelial nitric oxide synthase function in cerebral arterioles in vivo. AB - The marked impairment in cerebrovascular endothelial nitric oxide synthase (eNOS) function that develops after ovariectomy may relate to the observation that the abundance of cerebral vascular eNOS and its endogenous inhibitor, caveolin-1, vary in opposite directions with chronic changes in estrogen status. The authors endeavored, therefore, to establish a link between these correlative findings by independently manipulating, in ovariectomized female rats, eNOS and caveolin-1 expression, while monitoring agonist (acetylcholine)-stimulated eNOS functional activity. In the current study, the authors showed that individually neither the up-regulation of eNOS (through simvastatin treatment), nor the down-regulation of caveolin-1 (through antisense oligonucleotide administration) is capable of restoring eNOS function in pial arterioles in vivo in these estrogen-depleted rats. Only when eNOS up-regulation and caveolin-1 down-regulation are combined is activity normalized. These results establish a mechanistic link between the estrogen-associated divergent changes in the abundance of caveolin-1 and eNOS protein and eNOS functional activity in cerebral arterioles. PMID- 11487726 TI - Oxidative stress-dependent release of mitochondrial cytochrome c after traumatic brain injury. AB - Mitochondrial cytochrome c translocation to the cytosol initiates the mitochondrial-dependent apoptotic pathway. This event has not been previously reported in traumatic brain injury (TBI). The authors determined the expression of cytochrome c in cytosolic and mitochondrial fractions after severe TBI produced by the controlled cortical impact model in the mouse. One hour after trauma there was an increase in cytosolic cytochrome c immunoreactivity. The increases in cytosolic cytochrome c preceded DNA fragmentation, which started at 4 hours. Western blots of mitochondrial and cytosolic fractions confirmed that there was a translocation of cytochrome c from the mitochondria after TBI. Mice deficient in manganese superoxide dismutase (MnSOD) showed an increased loss of mitochondrial cytochrome c after trauma, but less apoptotic cell death 4 and 24 hours after injury compared with wild-type control mice. However, the overall cell death was increased in MnSOD mice, as illustrated by a larger cortical lesion in these animals. The results show that cytochrome c is released from the mitochondria after severe TBI partly by a free radical-dependent mechanism, and that massive mitochondrial cytochrome c release is a predictor of necrotic cell death rather than apoptosis. PMID- 11487727 TI - Neuronal, but not microglial, accumulation of extravasated serum proteins after intracerebral hemolysate exposure is accompanied by cytochrome c release and DNA fragmentation. AB - Vasogenic edema after oxidative injury has been accompanied by intracellular accumulation of serum proteins and nuclear damage. This study sought to determine whether serum protein accumulation, along with other markers of brain injury, was present after exposure to intracerebral hemolysate, an oxidant model of intracerebral hemorrhage (ICH). Saline (n = 24) or hemolysate (n = 30) was injected into the caudate-putamen of adult Sprague-Dawley rats. Compared with saline, hemolysate deposition was associated with intracellular accumulation of serum proteins as evidenced by Evans blue uptake in neurons and microglia at 4 and 24 hours. Intracellular Evans blue colocalized with DNA fragmentation detected by nick end-labeling and whose presence was confirmed by gel electrophoresis. Immunoblots of cytosolic fractions confirmed cytochrome c release. Immunostaining established colocalization of cytosolic cytochrome c and intracellular Evans blue at 4 hours. At 24 hours, cytosolic cytochrome c was evident in astrocytes surrounding Evans blue-positive cells. Immunoblot analysis and immunostaining revealed HSP70 induction at 24 hours in regions adjacent to intracellular serum accumulation. Neuronal accumulation of extravasated serum proteins in this model of ICH was associated with cytochrome c release, DNA fragmentation, and cell death. Stress protein induction in adjacent regions suggested that vasogenic edema might have exacerbated cellular dysfunction and cell death after ICH. PMID- 11487728 TI - Cloned blood-brain barrier adenosine transporter is identical to the rat concentrative Na+ nucleoside cotransporter CNT2. AB - Adenosine transport into brain is regulated by the activity of the adenosine transporter located at the brain capillary endothelial wall, which forms the blood-brain barrier (BBB) in vivo. To facilitate cloning of BBB adenosine transporters, poly A+ RNA was purified from isolated rat brain capillaries for production of a rat BBB cDNA library in the pSPORT vector. The cloned RNA (cRNA) generated from in vitro transcription of this library was injected into frog oocytes followed by measurement of [3H]-adenosine uptake. After dilutional cloning, a full-length, 2905-nucleotide adenosine transporter cDNA, designated clone A-11, was isolated. The A-11 clone yielded [3H]-adenosine flux ratios of 400 to 500 after injection of cRNA in oocytes. The adenosine uptake was sodium dependent and insensitive to inhibition by S-(4-nitrobenzyl)-6-thioinosine (NBTI). The Km and Vmax of adenosine transport in the cRNA-injected oocytes were 23.1 +/- 3.7 micromol/L and 10.8 +/- 0.9 pmol/oocyte. min. The K0.5 for sodium was 2.4 +/- 0.1 mmol/L, with a Hill coefficient (n) of 1.06 +/- 0.07. DNA sequence analysis indicated the rat BBB A-11 adenosine cDNA was identical to rat concentrative nucleoside transporter type 2 (CNT2). The adenosine transporter activity of the rat BBB A-11 CNT2 clone is 50-fold more active than previously reported rat CNT2 clones. In summary, these studies describe the expression cloning of CNT2 from a rat BBB library and show that the pattern of sodium dependency and NBTI insensitivity of the cloned CNT2 are identical to patterns of adenosine transport across the BBB in vivo. These results suggest that BBB adenosine transport in vivo is mediated by CNT2, which would make CNT2 one of the few known sodium-dependent cotransporters that mediate substrate transport across the BBB in the blood to brain direction. PMID- 11487729 TI - Interactions between the endothelium-derived relaxing factor/nitric oxide system and the endogenous opiate system in the modulation of cerebral and spinal vascular CO2 responsiveness. AB - The role of the L-arginine-nitric oxide (NO) system, the role of the endogenous morphine-like substances (endorphins), and the possible interaction between these two systems in the modulation of regional cerebral and spinal CO2 responsiveness was investigated in anesthetized, ventilated, normotensive, normoxic cats. Regional cerebral blood flow was measured with radiolabeled microspheres in hypocapnic, normocapnic, and hypercapnic conditions in nine individual cerebral and spinal cord regions. General opiate receptor blockade by 1 mg/kg naloxone intravenously alone or NO synthase blockade by 3 mg/kg N(omega)-nitro-L-arginine methyl ester (L-NAME) intravenously alone caused no changes in regional CO2 responsiveness. Combined administration of these two blocking agents in the very same doses, however, resulted in a strong potentiation, with a statistically significant reduction of the CO2 responsiveness observed. Separation of the blood flow response to hypercapnia and hypocapnia indicates that this reduction occurs only during hypercapnia. Specific mu and delta opiate receptors were blocked by 0.5 mg kg(-1) IV beta-funaltrexamine and 0.4 mg kg(-1) IV naltrindole, respectively. The role of specific mu and delta opiate receptors in the NO-opiate interaction was found to be negligible because neither mu nor delta receptor blockade along with simultaneous NO blockade were able to decrease CO2 responsiveness. The current findings suggest a previously unknown interaction between the endothelium-derived relaxing factor/nitric oxide (EDRF/NO) system and the endogenous opiate system in the cerebrovascular bed during hypercapnic stimulation, with the phenomenon not mediated by mu or delta opiate receptors. PMID- 11487730 TI - Ornithine decarboxylase knockdown exacerbates transient focal cerebral ischemia induced neuronal damage in rat brain. AB - Transient cerebral ischemia leads to increased expression of ornithine decarboxylase (ODC). Contradicting studies attributed neuroprotective and neurotoxic roles to ODC after ischemia. Using antisense oligonucleotides (ODNs), the current study evaluated the functional role of ODC in the process of neuronal damage after transient focal cerebral ischemia induced by middle cerebral artery occlusion (MCAO) in spontaneously hypertensive rats. Transient MCAO significantly increased the ODC immunoreactive protein levels and catalytic activity in the ipsilateral cortex, which were completely prevented by the infusion of antisense ODN specific for ODC. Transient MCAO in rats infused with ODC antisense ODN increased the infarct volume, motor deficits, and mortality compared with the sense or random ODN-infused controls. Results of the current study support a neuroprotective or recovery role, or both, for ODC after transient focal ischemia. PMID- 11487731 TI - Transient cerebral ischemia increases tyrosine phosphorylation of the synaptic RAS-GTPase activating protein, SynGAP. AB - Cerebral ischemia results in activation of the mitogen-activated protein kinase pathway and increased tyrosine phosphorylation of proteins associated with postsynaptic densities (PSDs). The authors investigated the possible relation between these events by determining the effect of ischemia on tyrosine phosphorylation of the brain-specific, PSD-enriched, Ras-GTPase activating protein, SynGAP. Transient (15 minutes) global ischemia was produced in rats by 4 vessel occlusion and PSDs prepared from forebrains immediately after ischemia or at 20 minutes, 1 hour, or 24 hours of reperfusion. Tyrosine phosphorylation of SynGAP was elevated relative to sham-operated controls by 20 minutes of reperfusion and remained elevated for at least 24 hours. Tyrosine phosphorylation of SynGAP also increased in CA1 and CA3/DG subfields of the hippocampus. Enhanced tyrosine phosphorylation of SynGAP was not accompanied by a change in PSD RasGAP activity. SynGAP bound to the SH2 domains of Src and Fyn in a tyrosine phosphorylation-dependent fashion, and this interaction increased after ischemia. SynGAP binds to the PDZ domains of PSD-95/SAP90 and coimmunoprecipitated with PSD 95. The coimmunoprecipitation of SynGAP with PSD-95 decreased after ischemia. The results indicate that changes in the properties and interactions of SynGAP may be involved in the neuropathology of ischemia. PMID- 11487732 TI - Delayed rt-PA treatment in a rat embolic stroke model: diagnosis and prognosis of ischemic injury and hemorrhagic transformation with magnetic resonance imaging. AB - The authors characterized effects of late recombinant tissue plasminogen activator (rt-PA) administration in a rat embolic stroke model with magnetic resonance imaging (MRI), to assess potential MRI correlates, or predictors, or both, of rt-PA-induced hemorrhage. Diffusion-, perfusion-, and postcontrast T1 weighted MRI were performed between 4 and 9 hours and at 24 hours after embolic stroke in spontaneously hypertensive rats. Treatment with either rt-PA or saline was started 6 hours after stroke. A spectrophotometric hemoglobin assay quantified hemorrhage severity. Before treatment, relative cerebral blood flow index (rCBFi) and apparent diffusion coefficient (ADC) in the ischemic territory were 30% +/- 23% and 60% +/- 5% (of contralateral), respectively, which increased to 45% +/- 39% and 68% +/- 4% 2 hours after rt-PA. After 24 hours, rCBFi and ADC were 27% +/- 27% and 59 +/- 5%. Hemorrhage volume after 24 hours was significantly greater in rt-PA-treated animals than in controls (8.7 +/- 3.7 microL vs. 5.1 +/- 2.4 microL, P < 0.05). Before rt-PA administration, clear postcontrast T1-weighted signal intensity enhancement was evident in areas of subsequent bleeding. These areas had lower rCBFi levels than regions without hemorrhage (23% +/- 22% vs. 36% +/- 29%, P < 0.05). In conclusion, late thrombolytic therapy does not necessarily lead to successful reperfusion. Hemorrhage emerged in areas with relatively low perfusion levels and early blood brain barrier damage. Magnetic resonance imaging may be useful for quantifying effects of thrombolytic therapy and predicting risks of hemorrhagic transformation. PMID- 11487733 TI - Targeting expression of hsp70i to discrete neuronal populations using the Lmo-1 promoter: assessment of the neuroprotective effects of hsp70i in vivo and in vitro. AB - Transgenic technology provides a powerful means of studying gene regulation and specific gene function with complex mammalian systems. In this study, the authors exploited the specific and discrete neuronal expression pattern mediated by promoter 1 of the Lmo-1 gene to study the neuroprotective effects of the inducible form of heat shock protein 70kD (hsp70i) in primary hippocampal cultures in a mouse model of global cerebral ischemia. Targeting expression of hsp70i to hippocampal neurons protected these cells significantly from toxic levels of glutamate and oxidative stress (for example, exposure to 10 micromol/L free iron produced a 26% increase in lactate dehydrogenase release from neurons cultured from wild-type mice, but a 7% increase in neurons cultured from hsp70i transgenic mice). Bilateral carotid occlusion (25 minutes) produced significantly less neuronal damage in the caudate nucleus and posterior thalamus in hsp70i transgenic mice than in wild-type littermates (for example, 21% +/- 9.3% and 12.5% +/- 9.0% neuronal damage in lateral caudate nucleus of wild-type and hsp70i transgenic mice, respectively, P < 0.05). The current study highlights the utility of targeted expression of transgenes of interest in cerebral ischemia and demonstrates that expression of hsp70i alone is sufficient to mediate the protection of primary neurons from denaturing stress and that expression of human hsp70i in vivo plays crucial role in determining the fate of neurons after ischemic challenge. PMID- 11487734 TI - Quantification of neuroreceptors in living human brain. v. endogenous neurotransmitter inhibition of haloperidol binding in psychosis. AB - The half-inhibition concentration (IC50) of a drug indicates its ability to inhibit the binding of other ligands of a receptor. The authors used positron emission tomography to test the hypothesis that haloperidol's IC50 toward the binding of tracer N-[11C]methylspiperone ([11C]NMSP) in brain must be increased in patients in whom more dopamine is bound to receptors than in healthy volunteers. The IC50 of haloperidol was significantly elevated from 1.5 nmol/L in healthy volunteers and patients with bipolar disease without psychosis to 4.5 nmol/L in patients with schizophrenia or bipolar disease with psychosis. The higher IC50 values in psychosis are consistent with an 8-fold increased binding of dopamine and a 16-fold elevated concentration of synaptic dopamine in psychosis. At the 80% haloperidol blockade of the receptors, the calculated amount of neurotransmitter bound in the patients with psychosis declined to twice the value estimated in the nonpsychotic subjects, that is, 5 pmol cm(-3). PMID- 11487735 TI - The (18)F-fluorodeoxyglucose lumped constant determined in human brain from extraction fractions of (18)F-fluorodeoxyglucose and glucose. AB - Quantification of regional cerebral glucose metabolism (CMRglc) using positron emission tomography and 18F-fluorodeoxyglucose (PET-FDG) requires knowledge of the correction factor between FDG and glucose net clearance, the FDG lumped constant (LC). Because diverging values for LC have been obtained, the authors reevaluated LC by measuring the ratio of the cerebral net extraction fractions of FDG (E*) and glucose (E) from arteriovenous cerebral measurements. Thirty subjects were studied (mean age = 25 +/- 4 years): 12 during a programed infusion of FDG and 18 after a bolus injection of FDG. In the infusion study, LC was calculated as the ratio E*/E. In the bolus study, E* was calculated from the slope of a Patlak-Gjedde plot. Lumped constant was significantly smaller in the infusion study as compared with the bolus study (0.48 +/- 0.16 vs. 0.81 +/- 0.27, P < 0.001). In 4 subjects studied during continuous FDG infusion for 2.5 hours, LC decreased to 0.36 +/- 0.11. These results suggest that the "steady-state" method underestimates LC because E* continues to decline because of significant labeled product. Further, the authors provide evidence for resetting of LC toward a greater value. The subsequent resetting of CMRglc provides a physiologically more meaningful estimate and allows for comparison of CMRglc values between different methodologies. PMID- 11487736 TI - Using (10)CO2 for single subject characterization of the stimulus frequency dependence in visual cortex: a novel positron emission tomography tracer for human brain mapping. AB - Carbon-10-labeled carbon dioxide ((10)CO2) with a half-life of 19.3 seconds offers almost ideal characteristics as a positron emission tomography (PET) tracer for assessment of the regional cerebral blood flow (rCBF) distribution, enabling multiple independent measurements at short intervals. To appraise the feasibility of (10)CO2 for localizing and characterizing human brain function in single subjects, the authors chose a well-characterized activation paradigm. In 6 healthy volunteers, 50 to 64 independent PET scans of the rCBF distribution were acquired while viewing an annular reversing checkerboard presented at 10 reversal frequencies between 0.03 and 30 Hz. Changes in regional cerebral activity as a function of reversal frequency were modeled in every subject using a set of polynomial basis functions, which, as predicted, showed highly significant second or third order relations located in the striatal cortex. Correlation coefficients (R2) ranged from 0.46 to 0.63. The average intersubject maximal response relative to the 0.03 Hz condition was 8.0% +/- 1.7% SD occurring at stimulus contrast reversal frequencies between 6 and 15 Hz with an average of 11.8 +/- 3.8 (SD) Hz. From the qualitative and quantitative replication of previous results it is concluded that (10)CO2 PET is a feasible technique for human brain mapping studies and a great improvement compared with the existing oxygen-15-labeled water (H(2)(15)O) PET method, particularly for single subject studies and parametric design. PMID- 11487737 TI - Career development and compensation: strategies for physicians in academic health science centers. A perspective from a Canadian academic health science center. PMID- 11487738 TI - Dietary supplements for attention-deficit/hyperactivity disorder--a fishy business? PMID- 11487739 TI - Searching for glycemic control in pediatric type 1 diabetes: A long way to go. PMID- 11487740 TI - Yellow mice, red hair, and childhood obesity: the melanocortinergic pathway in energy homeostasis. PMID- 11487741 TI - Hypothyroxinemia in the preterm infant: the benefits and risks of thyroxine treatment. PMID- 11487742 TI - A randomized, double-blind, placebo-controlled trial of docosahexaenoic acid supplementation in children with attention-deficit/hyperactivity disorder. AB - OBJECTIVE: To determine whether docosahexaenoic acid (DHA) supplementation for 4 months decreases the symptoms of attention-deficit/hyperactivity disorder (ADHD). STUDY DESIGN: Sixty-three 6- to-12-year-old children with ADHD, all receiving effective maintenance therapy with stimulant medication, were assigned randomly, in a double-blind fashion, to receive DHA supplementation (345 mg/d) or placebo for 4 months. Outcome variables included plasma phospholipid fatty acid patterns, scores on laboratory measures of inattention and impulsivity (Test of Variables of Attention, Children's Color Trails test) while not taking stimulant medication, and scores on parental behavioral rating scales (Child Behavior Checklist, Conners' Rating Scale). Differences between groups after 4 months of DHA supplementation or placebo administration were determined by analysis of variance, controlling for age, baseline value of each outcome variable, ethnicity, and ADHD subtype. RESULTS: Plasma phospholipid DHA content of the DHA supplemented group was 2.6-fold higher at the end of the study than that of the placebo group (4.85 +/- 1.35 vs 1.86 +/- 0.87 mol % of total fatty acids; P <.001). Despite this, there was no statistically significant improvement in any objective or subjective measure of ADHD symptoms. CONCLUSION: A 4-month period of DHA supplementation (345 mg/d) does not decrease symptoms of ADHD. PMID- 11487743 TI - Predictors of glycemic control and short-term adverse outcomes in youth with type 1 diabetes. AB - OBJECTIVES: To examine predictors of glycemic control and to assess how glycemic control affects the incidence of short-term adverse outcomes in a pediatric population with type 1 diabetes. STUDY DESIGN: Three hundred youth, aged 7 to 16 years, with type 1 diabetes who were receiving diabetes specialty care were followed up prospectively for 1 year. Treatment plans and frequency of adverse outcomes were ascertained by questionnaires and medical record review. Incidence rates of adverse outcomes were compared among 3 strata of the population, representing tertiles of baseline glycosylated hemoglobin (HbA1c). RESULTS: Blood glucose monitoring frequency was the sole modifiable predictor of HbA1c (P <.0001). Overall incidence rate of hospitalization was 13 per 100 person-years, more than 3 times the rate in the general pediatric population and significantly higher in the upper HbA1c tertile compared with the other strata (P =.001). Rate of emergency department use was 29 per 100 person-years and did not differ significantly among tertiles. Incidence of severe hypoglycemia was 62 per 100 person-years and notably high even in those with poorest glycemic control. CONCLUSION: Despite improvements in diabetes care, the incidence of short-term adverse events in children with type 1 diabetes remains high, particularly in those with poorest glycemic control. PMID- 11487744 TI - Mutational analysis of melanocortin-4 receptor, agouti-related protein, and alpha melanocyte-stimulating hormone genes in severely obese children. AB - OBJECTIVE: To search for mutations in melanocortin pathway elements, that is, the melanocortin-4 receptor (MC4R ), agouti-related protein (AGRP ), and (alpha melanocyte-stimulating hormone (alpha MSH ) genes in children with severe obesity. STUDY DESIGN: Direct sequencing of the MC4R encoding sequence and single strand polymorphism conformation analysis of AGRP and alpha MSH genes were performed in 63 severely obese children. Polymerase chain reaction (PCR) assays of restriction fragment length polymorphism were used to assess the frequency of each newly discovered mutation in 283 non-obese control subjects. RESULTS: Four dominantly inherited, heterozygous, missense MC4R mutations (Val50Met, Ser58Cys, Ile102Ser, and Ile170Val) were identified in 4 unrelated children and none of the control subjects. Expression of the obese phenotype was variable in mutation positive family members. Clinical and laboratory features were similar in the obese children with and without an MC4R mutation. Two polymorphisms were detected in the AGRP -encoding sequence (a silent mutation in exon 1 and Ala67Thr in exon 2), with similar frequencies in the obese and control groups. No mutations were found in the alpha MSH gene. CONCLUSIONS: MC4R mutations may be a non-negligible cause of severe obesity in children with variable expression and penetrance. Mutations in AGRP and alpha MSH genes were not among the causes of obesity in our population. PMID- 11487745 TI - Congenital microcephaly: phenotypic features in a consecutive sample of newborn infants. AB - OBJECTIVE: We compared the prevalence of major and minor anomalies in a consecutive sample of newborn infants with congenital microcephaly with that among normocephalic infants. STUDY DESIGN: Head measurements from >19,000 liveborn infants at 1 hospital during the years 1991 and 1992 were reviewed. Infants whose head circumference was in the lowest quartile (n = 850) were remeasured by research assistants to identify all whose head circumference was 2 SD below the mean for gestational age; 106 infants with congenital microcephaly were identified. Infants with microcephaly (n = 65) and 294 infants in a control group were examined systematically for major malformations and minor physical features. RESULTS: Four (6.2%) of the 65 infants examined either had a major malformation or were considered dysmorphic. One of the 4 had a specific multiple malformation syndrome, and 1 dysmorphic infant had a rare metabolic defect. Overall, the infants with microcephaly did not have a higher frequency of minor anomalies. However, there was a higher frequency of frontal bossing, small chin, and short nose with anteverted nares, which was associated with small body size rather than microcephaly. CONCLUSIONS: Congenital microcephaly is infrequently accompanied by major malformations and occurs rarely as part of a recognizable syndrome. PMID- 11487746 TI - The subtle facial signs of prenatal exposure to alcohol: an anthropometric approach. AB - OBJECTIVES: The purpose was to demonstrate that an objective, multivariate case definition of fetal alcohol syndrome (FAS) and partial fetal alcohol syndrome (PFAS) can be derived by means of craniofacial anthropometry. STUDY DESIGN: Craniofacial measurements (n = 21) were taken of 100 individuals who had been exposed to alcohol before birth (41 FAS, 59 PFAS) and 31 members of a control group. Multivariate discriminant analysis was used to identify craniofacial measurements that best differentiated the 3 groups (FAS, PFAS, and control). RESULTS: Both the FAS and PFAS groups had a unique craniofacial phenotype that could be accurately discriminated from that of the control group. Stepwise discriminant analysis identified 6 craniofacial measurements that could differentiate individuals with and without prenatal alcohol exposure with 96% accuracy, 98% sensitivity, and 90% specificity. CONCLUSIONS: Individuals with FAS and PFAS have a distinctive facial phenotype that can be characterized anthropometrically. The phenotypic case definition could be used as a screening tool to identify individuals prenatally exposed to alcohol who do not exhibit a "classic" FAS phenotype but exhibit a more subtle craniofacial dysmorphia. PMID- 11487747 TI - Transport risk index of physiologic stability: a practical system for assessing infant transport care. AB - OBJECTIVES: To develop and validate a practical, physiology-based system for assessment of infant transport care. STUDY DESIGN: Transport teams prospectively collected data, before and after transport, from 1723 infants at 8 neonatal intensive care units (NICUs) from 1996 to 1997. We used logistic regression to derive a prediction model for mortality within 7 days of NICU admission and develop the Transport Risk Index of Physiologic Stability (TRIPS). We validated TRIPS for prediction of 7-day mortality, total NICU mortality (until discharge), and severe (> or =grade 3) intraventricular hemorrhage. RESULTS: TRIPS comprises 4 empirically weighted items (temperature, blood pressure, respiratory status, and response to noxious stimuli). TRIPS discriminated 7-day NICU mortality and total NICU mortality from survival with receiver operating characteristic areas of 0.83 and 0.76, respectively. There was good calibration across the full range of TRIPS scores and gestational age groups. Increase and decrease in TRIPS scores after transport were associated with increased and decreased mortality, respectively. The receiver operating characteristic area for TRIPS prediction of severe intraventricular hemorrhage was 0.74. Addition of TRIPS improved performance of prediction models in which gestational age and baseline population risk variables were used. CONCLUSIONS: TRIPS is validated for infant transport assessment. PMID- 11487748 TI - Low systemic blood flow and hyperkalemia in preterm infants. AB - OBJECTIVE: Early low systemic blood flow is common in preterm infants. This study examines the relationship among low flow, renal function, and early changes in blood potassium (K(+)). METHODS: Preterm infants (n = 119) born before 30 weeks' gestational age underwent serial Doppler echocardiographic studies. Superior vena cava flow (SVC flow) was assessed as a measure of upper body systemic blood flow uncorrupted by systemic to pulmonary shunts. Serial whole blood K(+) concentrations on each arterial blood gas sample and urinary output in the first 48 hours were recorded. RESULTS: Most infants had a variable degree of rise in K(+) during the first 24 hours of life. The mean rate of rise was 0.17 mmol/L/h, the mean peak K(+) was 5.54 mmol/L, and the mean time of peak K(+) was 20 hours. The peak K(+) occurred after the lowest measured SVC flow in 84% of infants. A significant positive relationship was found between the lowest measured SVC flow and the mean (r = 0.31, P =.001) and peak (r = 0.31, P =.001) K(+) in the first 24 hours. Low SVC flow at 5 hours best predicted the rate of K(+) rise (r = 0.28, P =.002) and at 12 hours best predicted the peak K(+) concentration (r = 0.47, P <.001). The mean minimum SVC flow in the 17 babies who became hyperkalemic was 29.5 mL/kg/min versus 46.2 mL/kg/min in the 102 infants with normokalemia. Urine output in the first 24 hours was significantly lower in the hyperkalemic infants. A K(+) rate rise exceeding 0.12 mmol/L/h in the first 12 hours predicted low SVC flow with 93% accuracy. CONCLUSIONS: The data are consistent with a role for low systemic blood flow leading to reduced urinary output and subsequent hyperkalemia in preterm infants. PMID- 11487749 TI - Characterization of anorectal pressure and the anorectal inhibitory reflex in healthy preterm and term infants. AB - OBJECTIVES: To evaluate anorectal motor function in healthy premature and term infants with the use of micromanometric techniques. STUDY DESIGN: Anorectal manometry was performed in 22 healthy neonates (9 female) with a mean postmenstrual age of 32 weeks (range, 30 to 38 weeks) with a micromanometric anorectal assembly (od 2.0 mm). The assembly incorporated a 2-cm-long sleeve sensor for measurement of resting anal sphincter pressures and relaxation, and 4 sideholes recorded anal and rectal pressures. Rectal distension was performed with a latex balloon or direct air insufflation to elicit the anorectal inhibitory reflex (AR). RESULTS: The mean anal sphincter pressure, rectal pressure, and rhythmic wave frequency were 40 mm Hg (range, 7 to 65 mm Hg), 11 mm Hg (range, 1 to 27 mm Hg), and 10/min (range, 8 to 14/min), respectively. A normal AR could be elicited in 21 of the 22 infants studied. CONCLUSION: An anorectal micromanometric sleeve catheter is suitable for use in evaluating anorectal pressures in preterm and term neonates. Insufflation of air without the use of a balloon to elicit the AR is reliable and suitable for use in infants <34 weeks. Premature infants older than 30 weeks' postmenstrual age have normal anorectal pressures and a normal AR. PMID- 11487750 TI - Comparison of costs and referral rates of 3 universal newborn hearing screening protocols. AB - OBJECTIVE: To investigate the costs and referral rates of 3 universal newborn hearing screening programs: transient evoked otoacoustic emissions (TEOAE), automated auditory brainstem response (AABR), and a combination, two-step protocol in which TEOAE and AABR are used. STUDY DESIGN: Clinical outcomes (referral rates) from 12,081 newborns at 5 sites were obtained by retrospective analysis. Prospective activity-based costing techniques (n = 1056) in conjunction with cost assumptions were used to analyze the costs based on an assumed annual birth rate of 1500 births. RESULTS: Referral rates differed significantly among the 3 screening protocols (AABR, 3.21%; two-step, 4.67%; TEOAE, 6.49%; P <.01), with AABR achieving the best referral rate at discharge. Although AABR had the lowest referral rate at discharge and the highest pre-discharge costs, the total pre- and post-discharge costs per infant screened (AABR, $32.81; two-step, $33.05; TEOAE, $28.69) and costs per identified child (AABR, $16,405; two-step, $16,527; TEOAE, $14,347) were similar among programs. CONCLUSION: Although AABR incurs higher costs during pre-discharge screening, it has lower referral rates than either the TEOAE or two-step program. As a result, the total costs of newborn hearing screening and diagnosis are similar among the 3 methods studied. PMID- 11487751 TI - Home apnea monitor use in preterm infants discharged from newborn intensive care units. AB - PURPOSE: To identify current factors associated with home apnea monitor use in preterm infants and to determine whether home monitor use was associated with a shorter length of hospital stay. SETTING: We evaluated neonates who were < or =34 weeks' estimated gestational age and admitted for neonatal intensive care. We excluded neonates with congenital anomalies, neonates transferred out before discharge, and neonates who died. METHODS: Using a database created with a computer-assisted tool that generates hospital notes, we reviewed the epidemiology of monitor use. Differences between neonates sent home with an apnea monitor and those who were not were evaluated by using stepwise logistic regression analysis to identify which factors were independently associated with a neonate being discharged with a monitor. RESULTS: We studied 14,532 neonates; 1588 (11%) were sent home with monitors and 12,944 (89%) were not. The most important variables associated with being discharged with a monitor were site of care and a diagnosis of apnea. Site variation remained significant when adjusted for gestational age, diagnosis of apnea, and a history of use of methylxanthines. When corrected for gestational age, monitor use was not associated with shorter hospital stays. CONCLUSION: The data suggest that monitor use is more dependent on physician preference than medical indication and is not associated with earlier hospital discharge. PMID- 11487752 TI - Reliability and validity of the Harris Infant Neuromotor Test. AB - OBJECTIVE: To examine the reliability and validity of the Harris Infant Neuromotor Test (HINT), a screening tool that can be administered and scored in <30 minutes, with the goal of identifying neuromotor differences in infants aged 3 to 12 months. STUDY DESIGN: Infants, aged 3 to 12 months (n = 54), were assessed in 2 high-risk infant follow-up programs in Vancouver, British Columbia. Inter-rater, test-retest, and intra-rater reliability were examined. Concurrent and predictive validity of the HINT with the Bayley Scales of Infant Development II (BSID-II) were evaluated by using the Pearson product-moment correlation. RESULTS: Intraclass correlation coefficients for reliability for the Total HINT Score ranged from 0.98 to 0.99. Concurrent validity of the HINT with the BSID-II Mental Scale during the first year was r = -0.73 (P <.01), and with the BSID-II Motor Scale, r = -0.89 (P <.01). The predictive relationships between the HINT and the BSID-II at 17 to 22 months were r = -0.11 for the BSID-II Mental Scale and r = -0.49 (P <.01) for the BSID-II Motor Scale. CONCLUSIONS: The HINT is reliable for screening infant neuromotor performance and has strong concurrent validity with the Bayley-II Mental and Motor Scales. HINT scores during the first year accounted for 24% of the variance of Bayley-II Motor scores at 17 to 22 months. PMID- 11487753 TI - A randomized trial of two levels of iron supplementation and developmental outcome in low birth weight infants. AB - OBJECTIVES: To investigate the effect of increased iron intakes on hematologic status and cognition in low birth weight infants. STUDY DESIGN: We randomly assigned 58 infants to receive formula with 13.4 mg iron/L (normal iron) or 20.7 mg iron/L (high iron). At baseline, discharge, and at 3, 6, 9, and 12 months' corrected age, we assessed anthropometry; infections; red blood cell hemoglobin, catalase, glutathione peroxidase, red blood cell fragility (hydrogen peroxide test), and superoxide dismutase values; plasma malondialdehyde, ferritin, iron, transferrin, zinc and copper levels; and diet intake. Griffiths' Development Assessment was done at 3, 6, 9, and 12 months only. RESULTS: No statistical differences (P <.05) were noted for weight, catalase or malondialdehyde levels, red blood cell fragility, or Griffith's Development Assessment. Iron intakes were greater in the high iron group except at 12 months. Hemoglobin (high iron, 123 +/ 9; normal iron, 118 +/- 8) was not different at 3 months (P =.07). Plasma zinc levels (high iron, 70 +/- 14; normal iron, 89 +/- 27) and copper levels (high iron, 115 +/- 26; normal iron, 132 +/- 27; P =.06) at 12 months suggested inhibition of absorption by high iron formula. Glutathione peroxidase levels were higher in the high iron group. The total number of respiratory tract infections was greater in the high iron group (3.3 +/- 0.9) than in the normal iron group (2.5 +/- 0.9). CONCLUSION: In terms of cognitive outcome, there is no advantage associated with elevated iron intake for low birth weight infants. PMID- 11487754 TI - Breast-feeding and the risk of bronchial asthma in childhood: a systematic review with meta-analysis of prospective studies. AB - BACKGROUND: The protective effect of breast-feeding on the development of childhood asthma remains a matter of controversy. We conducted a systematic review of prospective studies that evaluated the association between exclusive breast-feeding during the first 3 months after birth and asthma. STUDY DESIGN: We searched the 1966-1999 MEDLINE database and reviewed reference lists of relevant articles to identify 12 prospective studies that met pre-stated inclusion criteria. Methodological aspects of the studies, duration and exclusivity of breast-feeding, and outcomes were assessed. Effect estimates were abstracted by the investigators, using a standardized approach. RESULTS: The summary odds ratio (OR) for the protective effect of breast-feeding was 0.70 (95% CI 0.60 to 0.81). The effect estimate was greater in studies of children with a family history of atopy (OR = 0.52) than in studies of a combined population (OR = 0.73). CONCLUSIONS: Exclusive breast-feeding during the first months after birth is associated with lower asthma rates during childhood. The effect, caused by immunomodulatory qualities of breast milk, avoidance of allergens, or a combination of these and other factors, strengthens the advantage of breast feeding, especially if a family history of atopy is present. PMID- 11487755 TI - Diagnosing pediatric asthma: validating the Easy Breathing Survey. AB - OBJECTIVE: To determine the sensitivity, specificity, and predictive value of a simple, self-administered questionnaire for the diagnosis of asthma in children. STUDY DESIGN: A questionnaire specifically designed to assist primary care providers in making a diagnosis of asthma in children was developed and administered in 4 different primary care and subspecialty clinics, validated, and then used as part of an asthma management program called Easy Breathing. Asthma diagnoses were made according to recommended National Asthma Expert Panel Guidelines. RESULTS: Four questions on the survey were shown to be sensitive and specific for asthma. The sensitivity was greater for all levels (mild, moderate, and severe) of persistent asthma than for mild, intermittent asthma. A positive response to any 1 of the 4 questions was over 94% sensitive for asthma; a negative response to all 4 questions was 55% specific for ruling out asthma. CONCLUSIONS: Patient responses to 4 specific respiratory symptom questions can assist primary care providers in diagnosing asthma in children. Primary care providers serving pediatric populations at high risk for asthma should consider asking patients or their parents these 4 questions regarding asthma symptoms on a regular basis. PMID- 11487756 TI - Predictors of hospitalization in children with acute asthma. AB - OBJECTIVE: To identify predictors of long duration of bronchodilator therapy in children with acute asthma. STUDY DESIGN: An emergency department prospective cohort study of 278 children > or =12 months of age, with clinical and socioeconomic parameters collected at baseline and 4 hours after administration of corticosteroids. Patients were classified into short and long therapy groups, with interval from first albuterol dose to initiation of administration every 4 hours < or =12 or >12 hours, respectively. Predictors significant by univariate analysis were examined by multiple logistic regression. RESULTS: Five variables were associated with long therapy (n = 85) versus short therapy (n = 193): previous intensive care unit admission (odds ratio [OR] 7.2, 95% CI = 1.85, 27.7); baseline oxygen saturation < or =92% (OR 2.6, 95% CI = 0.89, 7.4), asthma score > or =6/9 (OR 2.9, 95% CI = 1.9, 4.37), oxygen saturation < or =92% (OR 6.6, 95% CI = 1.34, 32.0), and hourly albuterol dosing interval (OR 4.3, 95% CI = 0.82, 22.12) 4 hours after administration of corticosteroids. Probability of long therapy was 91.8% to 99% for > or =3 predictors, but only 40.6% to 61.8% for individual factors. CONCLUSION: A combination of 3 or more factors predicts long bronchodilator therapy and signals the need for hospitalization. Children with only one predictor can be safely treated in the emergency department or observation unit and reevaluated. PMID- 11487757 TI - School attendance and school performance: a population-based study of children with asthma. AB - OBJECTIVE: To analyze school attendance and school achievement as outcomes of the care of children with asthma. METHODS: A previously identified Rochester, Minnesota, cohort of children with asthma and age- and sex-matched children without asthma were studied. School attendance, standardized achievement test scores, grade point average, grade promotion, and class rank of graduating students for children with asthma and control subjects were obtained from the Rochester Public School system. RESULTS: Children with asthma (n = 92) and age- and sex-matched non-asthmatic control subjects with 640 school-years of observation were studied. Children with asthma had 2.21 (95% CI, 1.41 to 3.01) more days absent than children without asthma. There was no significant difference in standardized achievement test scores (reading percentile difference 1.22% [95% CI, -3.68 to 6.12], mathematics percentile difference 2.36% [95% CI, 2.89 to 7.60], language percentile difference 2.96% [95% CI, -4.03 to 7.15]). There was no significant difference in grade point average, grade promotion, or class rank of graduating students. CONCLUSION: In this community, although children with asthma had 2 excess days of absenteeism, the school performance of children with asthma was similar to that of children without asthma. PMID- 11487758 TI - Responses of non-obese white children to treadmill exercise. AB - OBJECTIVE: To determine the response to maximal treadmill exercise with the Bruce protocol in a cohort of healthy non-obese American children and adolescents. STUDY DESIGN: A retrospective review of treadmill exercise studies on 347 white American children (188 boys, 159 girls) aged 5 to 18 years was performed with metabolic criteria to verify maximal exercise effort. Data on exercise endurance time, heart rate, blood pressure, and metabolic variables were assessed and compared between age groups and sexes by unpaired t testing and analysis of variance. RESULTS: Exercise endurance time was lower in all age groups when compared with earlier published data. Girls had a lower endurance time than boys at all ages. There was no significant difference in maximum heart rate with respect to age or sex. The maximum systolic blood pressure and diastolic blood pressure increased with increasing age in both boys and girls. Boys had significantly higher maximum systolic blood pressure and diastolic blood pressure after 13 years of age. The maximum absolute oxygen consumption and indexed oxygen consumption showed similar trends for both sexes. CONCLUSION: Lower exercise endurance times are seen despite physiologic evidence of maximal effort, raising the possibility that cardiovascular conditioning is reduced in contemporary American children. PMID- 11487759 TI - Survival after first esophageal variceal hemorrhage in patients with biliary atresia. AB - OBJECTIVE: To determine the influence of the new onset of esophageal variceal hemorrhage (EVH) on transplant-free survival in children with biliary atresia and to examine variables that predicted survival after the onset of EVH. METHODS: Retrospective chart review of 134 patients with biliary atresia who underwent portoenterostomy between 1973 and 1992 at a single institution; 29% had EVH. RESULTS: The risk of death or need for liver transplantation was 50% at 6 years after the initial episode of EVH. Patients with a serum bilirubin concentration < or =4 mg/dL at the first episode of EVH had transplant-free survival of >80% for 4 years after this episode, those with bilirubin levels >4 to 10 mg/dL had 50% survival at 1 year, and those with bilirubin levels >10 mg/dL had 50% survival at 4 months. The risk of death or transplant for a child with EVH and total serum bilirubin levels >10 mg/dL was 12.0 (95% CI: 6.0, 24.1), 4 to 10 mg/dL was 7.2 (3.1, 16.7), and < or =4 mg/dL was 0.6 (0.1, 3.1) times the risk of a same-aged child who did not have EVH. CONCLUSIONS: Children with biliary atresia and first EVH episode have a variable prognosis related to total serum bilirubin concentration at the time of the episode. PMID- 11487760 TI - Childhood vaccination against varicella: persistence of antibody, duration of protection, and vaccine efficacy. AB - OBJECTIVE: To document the duration of protection afforded by Oka/Merck varicella vaccine over a 7-year period. STUDY DESIGN: The subjects were healthy children 1 to 12 years of age originally enrolled in clinical studies to evaluate the primary immune response to varicella vaccine 6 weeks after vaccination. Each was monitored for antibody persistence, breakthrough infection, and household exposure to varicella to produce estimates of vaccine efficacy. RESULTS: The 6 year cumulative varicella antibody persistence rate was 99.5% (95% CI: 98.9%, 100.0%). The annual breakthrough rate through 7 years ranged from 0.2% to 2.3% per year; the estimated cumulative event rate was 6.5%. Comparison of the observed average annual breakthrough rate with the age-adjusted expected annual incidence rate of varicella in unvaccinated children corresponded to an estimated vaccine efficacy of 93.8% to 94.6%. Eighty vaccinated children were exposed to varicella in the household, resulting in 8 (10%) cases of infection. When compared with the historical attack rate of 86.8% in unvaccinated susceptible persons exposed to varicella in the household, this yields an estimated vaccine efficacy of 88.5% (95% CI: 80.9%, 96.1%). Varicella cases in vaccinated children generally were mild. CONCLUSION: The live attenuated varicella vaccine is highly effective in inducing persistent immunity and long-term protection against breakthrough varicella infection. PMID- 11487761 TI - Immunization of HIV-infected children with varicella vaccine. AB - OBJECTIVE: To determine the safety and immunogenicity of varicella vaccine in children with human immunodeficiency virus (HIV) infection. Children (n = 41) who were mildly affected by HIV (Centers for Disease Control and Prevention stage N1 or A1) and had no history or serum antibody indicative of prior varicella infection were immunized with two doses of live attenuated varicella vaccine. RESULTS: A minority of the vaccine recipients had mild local or systemic reactions. Vaccination had no effect on the clinical stage of HIV or the HIV RNA plasma load. CD4 cell percentage and CD4 cell count were marginally decreased at week 4 after the first vaccination; this effect was no longer present at week 8 after vaccination. Two months after the second dose of vaccine, 60% of vaccine recipients had anti-varicella antibody in their serum, and 83% had a positive lymphocyte proliferation assay response to varicella antigen. CONCLUSION: On the basis of its safety and immunogenicity, varicella vaccine should be considered in the childhood vaccines given to mildly affected HIV-infected children. PMID- 11487762 TI - Pediatric workforce: data from the American Board of Pediatrics. PMID- 11487763 TI - Benzyl alcohol, kernicterus, and unbound bilirubin. AB - A prospective study in 1983 demonstrated no significant relationship between serum unbound bilirubin levels and kernicterus. The presence of benzoate (a bilirubin-binding competitor) in the serum along with sample dilution, however, may have rendered the unbound bilirubin measurements in that study inaccurate. PMID- 11487764 TI - The prognostic value of ductal plate malformation and other histologic parameters in biliary atresia: an immunohistochemical study. AB - Ductal plate malformation, a term given to intrahepatic bile ducts that retain the fetal configuration, is observed in some cases of biliary atresia. We examined 21 livers from patients with biliary atresia; ductal plate malformation occurred in 38% of cases, and its presence was predictive of poor clinical outcome (P =.04). PMID- 11487765 TI - Hypoglycemia associated with clonidine testing for growth hormone deficiency. AB - We have observed 4 cases of hypoglycemia associated with clonidine stimulation of growth hormone secretion; only one patient had growth hormone deficiency. Significant drowsiness after administration of clonidine may prolong the period of fasting in these children and mask early signs and symptoms, leading to severe hypoglycemia. PMID- 11487766 TI - Incomplete carbohydrate absorption from fruit juice consumption after acute diarrhea. AB - We carried out a double-blind, randomized study in 60 children with acute diarrhea to determine their capacity to tolerate commonly consumed fruit juices. Feedings of juice with high fructose/glucose ratios and sorbitol resulted in incomplete carbohydrate absorption and recurrence of diarrhea during the phase immediately after improvement of the illness. PMID- 11487767 TI - Major advances in pediatric cardiology in the 20th century: II. Therapeutics. PMID- 11487768 TI - Serum amino acid concentrations in preterm infants. PMID- 11487770 TI - Rickets resurgence in the United Kingdom: improving antenatal management in Asians. PMID- 11487772 TI - Concerns about Ritalin. PMID- 11487774 TI - Benefit of intravenous immunoglobulin in autoimmune stiff-person syndrome in a child. PMID- 11487775 TI - Knowledge and management of malaria in under five children by primary health care workers in Ibadan South-east local government area. AB - The results of a survey of the knowledge and management practices of 61 health workers in five primary health care facilities in Ibadan South-east LGA are presented. In addition, 30 health workers were observed as they managed children with fever and the parasite status of 92 children diagnosed to have malaria was determined. Results revealed that 62(67.4%) children had the malaria parasite. Knowledge of some basic concepts was fairly adequate as the majority 46(75.4%) knew the cause of malaria. Treatment practices were poor as only 34(55.7010) and 39(63.9%) health workers respectively prescribed chloroquine and paracetamol correctly. Observation revealed that history taking and physical examinations were rudimentary. Scores out of 100 on correct prescriptions of chloroquine and paracetamol were 60.1 and 76.8 respectively. There is an urgent need for periodic education programmes, especially for health workers with many years of experience to help them maintain clinical skills and refresh their knowledge. PMID- 11487776 TI - Evaluation of nucleolar organizer regions in tumours of the jaw bones. AB - Nucleolar organizer regions [NORS] are loops of DNA that transcribe to ribosomal RNA. They can be visualized as intranuclear black dots by histochemical staining with a colloid silver solution. Silver stained nucleolar proteins (AgNORs) were counted in a variety of jaw bone tumours. In osteosarcomas, the number of AgNORs was also quantified before and after chemotherapy. Malignant bone tumour cells possessed more than five small AgNORs (5.54 +/- 0.44). Nuclei of benign jaw bone tumour cells had less than three (2. 97 +/-0.61). A significant difference in the number of AgNORs between osteosarcoma before chemotherapy (5.76 +/- 0.50) and after chemotherapy (3.89 +/- 1.65) was observed. (P < 0.05). The number ofAgNORs in recurrent osteosarcoma, recurrent ameloblastic carcinoma and recurrent chondrosarcoma was much higher than in their respective primary lesion but without statistical significant difference. The results of the present study indicate that the AGNOR count might help in determining malignancy, evaluating the effect of chemotherapy, and deciding the prognosis. PMID- 11487777 TI - Maternal mortality at Sagamu, Nigeria--a ten year review (1988 - 1997). AB - A review of maternal deaths at the Ogun State University Teaching Hospital, Sagamu, Nigeria over a 10 year period is presented. During the period, there were 92 maternal deaths, those from abortion and ectopic pregnancy inclusive. The total deliveries were 5423 giving a maternal mortality ratio of 1700 per 100,000. Ruptured uterus was the most common cause followed by eclampsia, postpartum haemorrhage and complications of abortion in that order. Unbooked patients constituted about one third of the total (29. 1 %). Primipara and grandmultipara were the most at risk of maternal death and the risk of dying following operative delivery was six times that of vaginal delivery. Easy access to affordable antenatal care, good blood transfusion services, more widespread use of contraceptives and training of traditional birth attendants would help reduce the risk of maternal death. PMID- 11487778 TI - Hypothalamic neuropeptide Y mRNA in pregnant, lactating and suckling rats. AB - Blood glucose, plasma insulin and luteinizing hormone levels were studied in pregnant wistra rats and those in early and late stages of lactation. NPY mRNA was also measured in whole hypothalamic tissue of these rats which were either fed ad libitum or food deprived to 80% of the relative controls. When fed ad libitum, hypothalamic NPY mRNA was not significantly elevated in the pregnant rats (111 +/- 2.1%). By the 5th and 4th days of lactation the mRNA had increased progressively (141 +/- 4.7% of control, p<0.01; 186 +/- 9%, p<0.001) respectively. Blood glucose levels were unchanged in pregnancy and lactation, however, insulin levels dropped significantly by the ]4th day of lactation (control 322.3 +/- 3.2; lactating 298.6 +/- 4.8 pmol/l; p<0.05). Luteinizing hormone was significantly reduced in the lactating rats (control 2.2 +/- 0.21, lactating 0.81 +/- 0.2 ng/ml;p<0.05). In food restriction, NPY mRNA was increased moderately in the non-pregnant state and enormously in late lactation (non pregnant 157 +/- 21%, lactating 333 +/- 35%, p<0.001). In a lactation, blood glucose was unchanged while plasma insulin and LH were reduced to 20% and 50% of controls respectively (insulin: control 110.3 +/- 2.0; lactating 18.3 pmol/l; LH. control 1.3 +/- 0.1; lactating 0.59 +/- 0.4 ng/ml p<0.01). Orexigenic effect of hypothalamic NPY is possibly responsible for the hyperphagia in lactating. Food restriction and lactation had additive lowering effect on plasma insulin but an additive increase on hypothalamic NPYmRNA. NPY message may be partially responsible for the anovulatory effect of lactation. PMID- 11487779 TI - Spinal tuberculosis: a 15 year review at OAUTHC Ile-Ife. AB - A review of the hospital records over 15 years was made. The objective of the study was to describe the clinical presentation, management and outcome of spinal tuberculosis. Thirty-four patients were studied. There were 15(44.16%) males and 19 (55. 9%) females. Their age range was 2-80 years with mean (+/- SD) of 25.28 +/- 22.33 years. The occupations most commonly affected are Students (44.1%), Pre school Children (17.6%) and Traders (14.7%). Back pain (100%), weight loss (47.1%), paraparesis (47.1%), kyphotic spinal deformity (32.4%) and night sweats (29.4%) were common features. Only 26.5% patients had active pulmonary tuberculosis. All the vertebral segments were involved. The Lumbar spine was the most involved vertebral segment in 50% patients. Using the Westergren method the mean (+/- SD) Erythrocyte Sedimentation Rate (ESR) mm/hour at the initial diagnosis was 83.58 +/- 31.11 mm/hour whereas three months after the commencement of antituberculosis chemotherapy it was 30.06 +/- 11.96 mm/hour. All the patients were given multiple antituberculosis drugs therapy and spinal traction. Spinal support was offered when the patients became ambulant. At the end of two years follow-up, 94.1% patients were alive, while 5.9% patients died during the course of treatment due to overwhelming sepsis. Among the patients that were alive, 14.7% still had persistence of their neurological deficit ie. paraparesis. Ten percent of the patients were mobilised on crutches while 5.96% were confined to wheelchair. This study shows that in tuberculosis endemic areas, symptom of back pain especially in the younger age group should be thoroughly investigated as this group constituted the largest affected population. PMID- 11487780 TI - Giant congenital pigmented naevus with unusual presentation and early malignant transformation in a Nigerian infant: case report and review of the literature. AB - A six month-old female infant with a giant congenital pigmented naevus is reported because of the rarity and unusual presentation and early malignant transformation. It was the first case to be documented in our centre. There was an extensive giant, darkly pigmented naevus of the "bathing trunk" distribution noted at birth, associated with multiple darkly pigmented smaller naevi scattered over the trunk and limbs. Bv the third month of life large pigmented lesion began to demonstrate small papillomatous projections studding the entire surface and at age six months nodular and warty excrescences appeared including a large pedunculated spherical mass attached to the anal verge. The labia majora became anatomically disfigured by these nodular, warty lesions of various sizes associated with the large pigmented hairy naevus. Histological examination of the excised large perianal and vulvar nodules showed the typical picture of an intradermal naevus consistent with giant congenital pigmented (melanocytic) naevus. The deeper portions of the lesion showed features of malignant transformation. The rapidity of evolution of the lesions including histologic features of early malignant transformation and the gross disfiguring of the genitalia and perineum are uncommon presentation of this melanocytic naevus. In view of the early malignant transformation immediate surgical treatment was indicated. Unfortunately the patient was lost to follow-up. The case report is complimented with a review of the literature. PMID- 11487781 TI - Outcome of primary closure of incised acute soft-tissue abscesses. AB - One hundred and six patients with acute soft tissue abscesses were involved in the study at the Accident and Emergency unit of Jos University Teaching Hospital (JUTH) from June, 1996 to April, 1997. The objective of the study was to assess the outcome of primary closure in the treatment of acute soft tissue abscesses using the healing rate, cost effectiveness, quality of scars and postoperative complications as parameters. The patients were divided into two groups: group A for primary closure, and group B for the conventional method. There was a statistically significant difference (P<0.05) in the healing time for study group A (6.96 +/-123.56) compared with control group B. Similarly group A was more cost effective (Total cost = N406 +/-123.56) compared with control group B (Total cost = N580.47 +/- 174.24). The difference was statistically significant (P 3 mm). Operating time was not significantly lengthened, except in transphenoidal cases (255 +/- SD 168 min versus 185 +/- SD 119 min, p = 0.02). Length of stay was shorter in navigation cases in supratentorial glioma (12 +/- SD 7.3 days versus 15 +/- SD 7.9 days) and meningioma (11 +/- SD 5.3 days versus 16 +/- SD 6.7 days, p = 0.01). Outcome at 3 months was comparable in both the supratentorial glioma and meningioma group. Problems such as setup time and brain shift are addressed and discussed. PMID- 11487795 TI - Microsurgical anatomy of the Liliequist's membrane and surrounding neurovascular territories. AB - Since the original description by Liliequist only a few microanatomical studies of the Liliequist membrane have been performed. These studies contain some discrepancies in the description, boundaries, and attachments of the membrane. Using a surgical microscope the authors examined the microsurgical anatomy of Liliequist's membrane and surrounding neurovascular structures in twenty adult brains injected with silicone rubber, with special emphasis given to the analysis of controversial details. This description is intended as an aid for neurosurgeons performing neuroendoscopic procedures. PMID- 11487796 TI - An intramural macrocyst of an acoustic neurinoma rupturing after gamma knife radiosurgery: a case report. AB - We want to describe the rare case when an intramural macrocyst within an acoustic neurinoma (ACN) treated by gamma knife radiosurgery (GKRS) ruptured, followed by an impressive decrease of tumor volume and improvement of neurological symptoms. In a 59-year-old female patient, a large ACN with a hugh intramural macrocyst was diagnosed. As she refused open surgery, we performed GKRS covering the tumor margin and the cyst with 11 Gy. Seven months after treatment symptoms worsened slightly. Magnetic resonance imaging (MRI) revealed no significant change of tumor volume. One year after GKRS she felt a sensation behind her treated ear, followed by an immediate improvement of all her symptoms. Trigeminal hypaesthesia and vertigo disappeared, tinnitus ameliorated. A control MRI showed the cystic compartment no longer, the solid part shrunk within the following six years. Within the whole follow-up period hearing was stable. To our knowledge this is the first report of a macrocyst within an ACN to rupture after GKRS. PMID- 11487797 TI - Minimally invasive treatment of cavernous angioma of the optic chiasm: case report. AB - Cavernous angioma of the optic chiasm or optic nerve is extremely rare. We report the case of a 58-year-old woman with cavernous angioma of the optic chiasm. The lesion was totally removed through the eyebrow keyhole approach, which allowed appropriate intraoperative exploration of the optic chiasm and related structures. The present case confirms that a cavernous angioma located in the optic chiasm can be totally resected without further impairment of visual function. PMID- 11487798 TI - Multiple sclerosis with mass effect: case report. AB - Multiple sclerosis (MS) may rarely present as a cerebral mass with the clinical features and computed tomography (CT) scan appearance of a cerebral tumor. We report a case of MS with large cranial involvement showing a mass effect. We carried out a complete examination, including contrast enhancement, with neuroimaging studies. The operative procedure associated with medical treatment was performed and we obtained a good result. PMID- 11487799 TI - Rehabilitation for traumatic brain injury. PMID- 11487800 TI - Pharmacotherapy for alcohol dependence. PMID- 11487801 TI - Prevention and management of urinary tract infections in paralyzed persons. PMID- 11487802 TI - Treatment of depression--newer pharmacotherapies. PMID- 11487803 TI - Think small. PMID- 11487804 TI - A method to assess predominant energies of exposure in a nuclear research centre- Saclay (France). AB - A study of dosimetric errors is under way within an international collaborative study of cancer risk among workers in the nuclear industry. The objective is to quantify errors in the estimated photon doses to individual organs used for cancer risk estimation. One source of errors is the response of old dosemeters in workplace exposure conditions. As these conditions are not well known, the International Study must rely on expert estimations. This paper describes a method to assess the proportion of the dose from photons in three energy ranges (< 100, 100-300, > or = 300 keV) using the responses under filters of a multi element dosemeter. The method was tested on experimental and simulated data and provides a good estimate of the proportion of dose from photons below 100 keV, the most critical for dosemeter response. It was applied to personnel readings in one facility, confirming the experts' estimation. Beyond the International Study, the method has implication for the monitoring and protection of workers. PMID- 11487805 TI - The response of the sievert instrument in neutron beams up to 180 MeV. AB - Measurements with a tissue-equivalent proportional counter (TEPC) using the variance-covariance method have been performed in neutron beams between 71 keV and 180 MeV and in the cosmic radiation reference field (CERF) at CERN. The results show that with appropriate linear QD(yD) relations, the ambient dose equivalent can be determined within about 55% in these beams. Build-up measurements show that wall thickness is not crucial for H* determinations at 60 and 180 MeV. PMID- 11487806 TI - Extended survey of indoor and outdoor terrestrial gamma radiation in Greek urban areas by in situ gamma spectrometry with a portable Ge detector. AB - The results obtained from more than 1000 indoor and outdoor in situ gamma spectrometry measurements in 41 towns (from all geographic subdivisions) of the Greek mainland (not islands) are presented. From the in situ gamma spectra the absorbed dose rate in air due to uranium series, thorium series, 40K and 137Cs are derived and discussed. PMID- 11487807 TI - In vivo measurement of unattached radon progeny deposited in the human respiratory tract. AB - Seven nose breathing and seven mouth breathing volunteers were exposed to atmospheres enriched with unattached radon progeny (218Po, 214Pb and 214Bi). The activity of these radionuclides deposited in the respiratory tract was measured in vivo after the exposures. The results of these measurements are in agreement with predictions calculated with the ICRP Publication 66 Human Respiratory Tract Model. Temporal analysis of the activity deposited in the heads of the volunteers leads to the conclusion that a significant amount of the deposited activity associated with particle diameters of about 1 nm is not subject to a fast transport to the gastrointestinal tract as generally reported for larger aerosol particles. PMID- 11487808 TI - The long-term decrease of 90Sr availability in the environment and its transfer to man after a nuclear fallout. AB - Due to its long physical half-life, and the fact that its long-term mobility in the environment as well as its radiotoxicity is higher than that of 137Cs, the long-term bio-availability of 90Sr in the environment is of importance with regard to the long-term population exposure after fallout from nuclear weapons detonations or a severe reactor accident. It will also substantially influence the time-span required until re-utilisation of highly contaminated territory is possible again. An assessment of the long-term decrease of the activity concentration in all foodstuffs relevant for internal exposure after severe 90Sr fallout was performed. The observed effective half-lives were approximately 1.8 2.1 years in the first 2-3 years after the end of fallout and 8-10 years in the following three decades. This is equivalent to a biological half-life of about 13.2 years and results in a total 50 year dose of 6.2 times the first year exposure. Due to this decline in 90Sr-availability, the average annual activity intake of 90Sr in Austria has decreased from 840 Bq at the climax of the nuclear weapons tests to about 42 Bq in 1997 for adults, and from 500 Bq to about 35 Bq for 1 year old infants. This is equivalent to a 90Sr ingestion dose of 1.2 microSv for adults and 2.5 microSv for 1 year old infants in 1997 or less than 0.4% of the ingestion dose by natural radionuclides in the diet. PMID- 11487809 TI - Effect of absorption parameters on calculation of the dose coefficient: example of classification of industrial uranium compounds. AB - In the Human Respiratory Tract Model (HRTM) described in ICRP Publication 66, time-dependent dissolution is described by three parameters: the fraction dissolved rapidly, fr, and the rapid and slow dissolution rates sr and ss. The effect of these parameters on the dose coefficient has been studied. A theoretical analysis was carried out to determine the sensitivity of the dose coefficient to variations in the values of these absorption parameters. Experimental values of the absorption parameters and the doses per unit intake (DPUI) were obtained from in vitro dissolution tests, or from in vivo experiments with rats, for five industrial uranium compounds UO2, U3O8, UO4, UF4 and a mixture of uranium oxides. These compounds were classified in terms of absorption types (F, M or S) according to ICRP. The overall result was that the factor which has the greatest influence on the dose coefficient was the slow dissolution rate ss. This was verified experimentally, with a variation of 20% to 55% for the DPUI according to the absorption type of the compound. In contrast, the rapid dissolution rate sr had little effect on the dose coefficient, excepted for Type F compounds. PMID- 11487810 TI - Natural radionuclides contents and radon exhalation rates in building materials used in South Korea. AB - The natural radionuclide (226Ra, 232Th and 40K) contents of building materials commonly used in South Korea have been determined by a gamma ray spectroscopy system using a high purity germanium detector. The radon exhalation rates and emanation coefficients of building materials have also been studied. The average concentrations of 226Ra, 232Th and 40K in building materials were in the range of 6.47-271 Bq.kg-1, N.D.-89.9 Bq.kg-1 and 16.8-1081 Bq.kg-1, respectively. The average radon exhalation rates and emanation coefficients were in the range of 0.0061-0.2404 Bq.kg-1.h-1 and 2.39-11.5%, respectively. PMID- 11487811 TI - Responses of TLD-BeO:Na (UD-170A) to heavy ions and space radiation. AB - Responses of TLD-BeO:Na (UD-170A) to high-LET particles were examined with selected heavy ion beams (He, C, Ne, Ar, and Kr) at NIRS-HIMAC, and compared with TLD-Mg2SiO4:Tb (TLMS) and radiophotoluminescent glass (RPLG). The relative TL efficiency of UD-170A as 137Cs gamma ray equivalent arose notably with increasing LET infinity.H2O for He and C, and decreased for the heavier charged particles. In contrast, the efficiencies of TLMS and RPLG did not increase over the range of LET from 0.5 to 410 keV.micron-1. The three detectors were used for space radiation measurement in the Mir space station for 40 days at 400 km altitude and 51.65 degrees inclination. The values from each detector as gamma ray absorbed dose equivalent showed a large spatial variation by a factor 2 in the same Core module. The detector values were in the order of UD-170A > TLMS > RPLG as expected from the results obtained on the ground, although ratios of these values changed depending on positions. These results indicate that both radiation quality and dose level in a spacecraft change significantly and a measurement at one location cannot accurately represent the individual dose to an astronaut. These small detectors should be useful as supplementary personal dosemeters for astronauts. PMID- 11487812 TI - Short-lived alpha sources of energies 6.0 MeV and 7.69 MeV for calibration purposes. AB - An approach for preparation of short-lived alpha sources of energy 6.0 MeV and 7.69 MeV is proposed. The sources are prepared by taking a sample of 222Rn progeny on an alpha spectrometric filter. The activities (or related parameters) of 218Po, 214Pb and 214Bi at the end of sampling are precisely determined by a reference measurement with an alpha spectrometer. Further they are used as input values to calculate with a sufficient precision the number of emitted alpha particles of any energy and at any time interval of interest. Theoretical modelling and experimental results demonstrated that such sources could be prepared with a sufficient purity. There is a potential for the number of alpha particles emitted in a given time interval to be certified with an accuracy of 1 2%. PMID- 11487813 TI - In radiological protection, the protection quantities should be expressed in terms of measurable physical quantities. AB - It is somewhat of a truism that is expressed in the often quoted statement that you can't go home again. In the days when life was simpler, we could assume that a measurement performed with a portable survey instrument yielding a value for exposure rate in terms of roentgens/hour was a sufficient approximation to the value for the dose equivalent rate or absorbed dose rate at a point in a radiation field. As our understanding of radiological protection grew more sophisticated, we found it necessary to introduce additional rigor into the definitions of the radiation protection quantities. As mentioned earlier, we demand a lot from the definitions of these quantities. They need to satisfy many conditions, perhaps too many. When they are examined from the point of view of a metrologist, they may be considered to be deficient in many respects. When they are viewed by a radiobiologist, they may appear to be naive or overly simplistic. A radiation protection technologist at a nuclear power plant may have difficulty understanding their sometimes-subtle implications. The ICRU and ICRP have worked for quite some time to incorporate all the necessary requirements into the definitions of the operational and protection quantities. From the arguments of our debaters, one might conclude that after careful consideration of past developments, additional refinements will be devised in the future. PMID- 11487814 TI - Patient dosimetry protocols in digital and interventional radiology. AB - Dosimetry requirements and protocols for performing measurements in digital and interventional radiology are discussed. Calculated entrance surface dose (ESD) is predicted to be of increasing interest in the future, replacing direct measurement with thermoluminescence (TL) dosemeters. The quantities proposed for establishment of reference values for interventional radiology are reviewed briefly, and the methods of collecting the data required for estimation of their values by means of traditional manual and new automatic methods are compared. It is concluded that the manufacturers of X ray units can largely solve the dosimetry problems of interventional radiology in machines with fully digital control systems after they have received sufficient data on patient dosimetry requirements. PMID- 11487815 TI - Approaches to establishing reference levels in interventional radiology. AB - Reference levels in radiodiagnostics are a requirement stated by the Council Directive 97/43/EURATOM. Reference levels are also relevant for interventional procedures, in accordance with this Directive, which claims special attention to quality assurance programmes, including quality control and patient dose evaluations for special practices such as interventional radiology, to assess the convenience of corrective action depending on the measured doses. The present paper addresses a method of establishing reference levels in interventional radiology, in the framework of optimisation, discussing the ways of putting forward values with a degree of tolerance, to allow for procedure complexity, depending on patient pathology. The need for several estimators used together, namely fluoroscopy time, total number of images per procedure and dose-area product, is also emphasised, proposing a further skin dose estimate in case of risk of deterministic effects. Finally, a brief summary of principles is given for the correct management of reference levels in interventional radiology. PMID- 11487816 TI - Patient and staff dosimetry problems in interventional radiology. AB - Interventional radiology has developed into a dynamic part of radiology over the past twenty years, combining diagnostic and therapeutic methods. On the other hand, it is associated with high radiation doses to patient and staff, due to extended fluoroscopy times and the large number of radiographs. Also, occupational exposures from interventional radiology procedures have a tendency to be greater than other radiological examinations. The need for measuring and evaluating patient and staff doses is apparent. However, dose estimations depend on a large number of factors making these procedures very complex. The aim of this study is to review all the different approaches that appear in the literature on this matter, to delineate the different dosimetry protocols that are proposed and to focus on the practical problems that arise when an evaluation or comparison of dosimetry results is attempted. PMID- 11487817 TI - Paediatric dose reduction with the introduction of digital fluorography. AB - Fluoroscopy guided examinations in a paediatric X ray department were initially carried out on a unit that used a conventional screen-film combination for spot films. A new fluoroscopy unit was installed with the facilities of digital fluorography and last image hold. Comparison of equipment performance showed that the dose per image for screen-film and digital fluorography was 3 microGy and 0.4 microGy, respectively. Although the screen-film had superior image quality, the department's radiologist confirmed that digital fluorography provided a diagnostic image. Patient dose measurements showed that introduction of the new unit caused doses to fall by an average of 70%, although fluoroscopy time had not changed significantly. The new unit produced 40% less air kerma during fluoroscopy. The remaining 30% reduction in dose was due to the introduction of digital fluorography and last image hold facilities. It is concluded that the use of digital fluorography can be an effective way of reducing paediatric dose. PMID- 11487818 TI - Personnel doses in haemodynamic units in Greece. AB - Personnel of haemodynamic and interventional radiology units receive continuously increasing radiation doses due to extended fluoroscopy. Moreover, there is not a parallel increase in the number of cardiology specialists involved. Doses received by 15 cardiologists and 5 nurses, in 5 Athenian hospitals were measured using thermoluminescence dosemeters (TLD) and film badges. The workload and examination protocol of each cardiologist, the technical characteristics of the X ray unit, as well as availability and use of protective equipment were recorded. Results show that doses measured by TLD and film badges differ due mainly to the irregular wear of the latter. Although X ray units performed comparably with each other, dose per procedure received by each cardiologist varied widely, due to differences in examination protocol and beam collimation used. In all cases, nurses' dose was approximately one fourth of cardiologists' dose. In half of the cases assessed, the protective equipment available was not in full use. Estimation of dose per procedure, taking into account personnel's workload assesses annual personnel doses. Evaluation of risk level and check of compliance with regulatory dose limits should be part of continuing radiation protection education. PMID- 11487819 TI - Intercomparison of TL dosimetry (DIMOND programme). AB - During DIMOND CA, a thermoluminescence dosimetry intercomparison has been performed, in order to ensure that all participants measure the same dose. After the calibrating and annealing procedures, 50 TLD pellets (in groups of five) of each participating country were sent to the coordinator to be irradiated with 'unknown' radiations (blind test). The irradiation conditions (8 different set ups, one for each group of pellets per participant) were (i) one gamma irradiation (60Co), at air kerma in the range of 0.3 to 50.0 mGy, (ii) seven X ray irradiations, at air kerma in the range of 0.3 to 50.0 mGy, with tube potential in the range of 40 to 150 kV and a variety of filter combinations. Ten pellets (two groups) were used for background and transport dose evaluation. After measuring the TLDs, the participants sent the results to the coordinator for intercomparison. Irradiating set-up conditions, evaluated doses, as well as comparison of mean values with the 'true' nominal doses are presented and discussed. PMID- 11487820 TI - Occupational exposure in angiography (Prague workplaces). AB - The results of measurements, which were performed at angiography departments, are presented. The aim of this work was not only to assess patient exposure but also to perform a survey of the occupational exposures of physicians during common angiography examinations of the skull, chest, abdomen and limbs. Patient data, type of examination, fluoroscopy time, the whole number of film frames, dose-area product (Diamentor E) and physician dose (Stephen 6000) were also recorded. There are many factors influencing the level of measured exposures. They include input parameters of X ray performance (kV, mA, projection, diaphragms, ZOOM, type of record etc) and individual approach of the physicians. This paper contributes to the discussion about the increasing exposures of the physicians specialising in diagnostic and interventional angiography. PMID- 11487821 TI - Regulatory initiatives and framework in the USA: interventional radiology. AB - Therapeutic procedures using interventional radiology equipment as the imaging tool are often the only available treatment for serious, life-threatening conditions. Many of these procedures require extended periods of radiation exposure often at one location on the patient. As a result, the US Food and Drug Administration (FDA) continues to receive reports of radiation-induced injuries to the skin in patients undergoing some of these therapeutic procedures. The regulatory scheme applicable to interventional radiology equipment in the US is described including federal, state and local aspects, and the impact of professional and non-governmental organisations. Current activities within the FDA and other organisations which have or will affect the use, maintenance and manufacture of interventional radiology equipment are also described. PMID- 11487822 TI - Guidance levels in the Nordic countries: a preliminary report for selected interventional procedures. AB - The Nordic radiation protection authorities have already published recommended guidance levels for patient doses for six conventional radiological examinations. Over the past two years a similar protocol has been in progress for three interventional procedures. Measurements have been performed in 22 different hospitals in the Nordic countries on patients in the weight range 40-100 kg. The selected procedures are percutaneous transluminal coronary angioplasty (PTCA), percutaneous transluminal angioplasty (PTA) and endoscopic retrograde cholangio- and pancreatio-graphy (ERCP). A total of 281 PTCA procedures, 304 PTA procedures and 147 ERCP procedures are included in the study. The results from this survey are presented as a first attempt to set guidance levels. PMID- 11487823 TI - Training and accreditation in radiation protection for interventional radiology. AB - Training in radiation protection is a basic aspect of the optimisation of medical exposures. Council Directive 97/43/EURATOM establishes the need for an adequate theoretical and practical training of the staff working in radiological practices, and competence in radiation, for which Member States shall ensure the establishment of appropriate curricula. Keeping in mind the different specialities and professional responsibilities, training curricula must be proposed and endorsed to achieve a common core of knowledge in radiation protection throughout Europe, for different groups of health workers. In interventional radiology, previous initiatives led to the definition of a syllabus of educational objectives and to its testing in a specific course. The present paper presents educational objectives for interventional radiology, developed in the framework of the DIMOND European concerted action. PMID- 11487824 TI - Dose displays and record keeping. AB - In digital diagnostic and interventional radiology, patient dosimetry is essential. When considering the display requirements for interventionalists it is necessary to consider the basis for their information needs. An interventionalist must be made aware of the potential for deterministic and non-deterministic effects. Thus a display is required which provides an indication of whether skin effects are likely and this predicates a display of maximum estimated skin entrance dose. For non-deterministic effects, a display of dose-area product is useful. Unfortunately, neither display is generally available to the interventionalist, so inevitably they will remain in the dark about the potential radiological consequences of the procedure. Given the need to follow up patients, who may receive a high dose, it is important that the patient notes contain relevant dosimetry data. One way of achieving this in the future would be to link electronically the patient dosimetry device to the patient record in the Radiological Information System. This dictates a need to develop patient dosimetry record minimum dataset and equipment connectivity standards. PMID- 11487825 TI - Review of dosimetry instrumentation in digital and interventional radiology. AB - Some dosimetry instruments and products are reviewed, the main emphasis being on patient dosimetry, recommendations for accuracy in different measurement applications and the results of some intercomparisons. It seems to be a common problem that the users of the general purpose air kerma (Ka) meters, dose-area product (DAP) meters or products such as thermoluminescence (TL) dosemeters are not always able to select the correct ionisation chamber, the calibration factor of a DAP meter or the TL dosemeter material and type, respectively, for different radiation conditions. The combined DAP and Ka meters developed recently, as well as the exposure data acquisition systems designed for monitoring one or more quantities or for determining the effective dose of a complicated examination, are described briefly. The most advanced software of these systems is able to display the dose distributions for the most exposed areas of the skin, on-line. PMID- 11487826 TI - The calibration of dosimetry instruments used in interventional radiology. AB - No general agreement about the definition of the patient dose exists. As regards the radiation health risk, the doses to specific organs, Hi, are the ultimate measures for a patient dose. Values of the calibration measured, Hi, are provided only by calculational means. Out of the whole process of patient dose determination, the instruments to measure X ray spectra, FSD, field dimensions and Ka can be calibrated, X ray quality is derived from the total filtration and kV value. The actual dynamic and X ray quality ranges shall be considered when air kerma and DAP meters are calibrated. A DAP meter measurement averages the uniform radiation field specific for the X ray tube assembly used and for the beam shaping technique performed. Therefore, a DAP meter calibrated on site is preferable for patient dosimetry in interventional radiology. PMID- 11487827 TI - The use of dynamic phantoms in interventional radiology. AB - The authors have constructed a 2D motor-controlled test object phantom holder to simulate clinical situations in which patient movement could be a cause of image degradation. The PAtient MOvement SImulation Test Object (PAMOSITO) has been constructed with modular parts to use different mobile test objects and static structures. The system allows the programming of different cycles of movement along two axes. PAMOSITO has been used in X ray equipment dedicated to interventional radiology. Those systems usually allow for different values for frame rate, pulse width or weighted frame averaging methods. The influence of selecting different values of the parameters, patient movement and its relation to patient dose and image quality has been studied. Image blurring due to motion has been evaluated with Leeds test objects TO.10 and 18FG. Spatial resolution limits and the threshold contrast detail detectability performance have been studied. PMID- 11487828 TI - A new tool for benchmarking cardiovascular fluoroscopes. AB - This paper reports the status of a new cardiovascular fluoroscopy benchmarking phantom. A joint working group of the Society for Cardiac Angiography and Interventions (SCAI) and the National Electrical Manufacturers Association (NEMA) developed the phantom. The device has been adopted as NEMA standard XR 21-2000, 'Characteristics of and Test Procedures for a Phantom to Benchmark Cardiac Fluoroscopic and Photographic Performance' in August 2000. The test ensemble includes imaging-field geometry, spatial resolution, low-contrast iodine detectability, working thickness range, motion unsharpness, and phantom entrance dose. The phantom tests systems under conditions simulating normal clinical use for fluoroscopically guided invasive and interventional procedures. Test procedures rely on trained human observers. PMID- 11487829 TI - A method based on DIMOND quality criteria to evaluate imaging in diagnostic and interventional cardiology. AB - Image quality criteria (IQC) for cine-angiography were recently settled. The aim of this study was to test whether these criteria allow a measurement of the quality of cine-angiograms. A questionnaire was derived from IQC where a binary response was required regarding the degree of visibility of anatomic or pathologic structures. Scores were given on a ranking scale. Two quality scores were defined (total score and minimum score) and standard deviation (SD) was assumed to be an indicator of the method's reproducibility. Data of the total score are presented for the first nine angiograms. Six experts obtained thirty nine readings. The total scores ranged between 83 and 99% (SD 0.8-18.7%); 89% of the readings were within 4% of SD. This preliminary experience indicates that quality criteria can be translated into a scoring system that yields reproducible data in most instances. The analysis of the remaining angiograms will help in understanding how to improve these results. PMID- 11487830 TI - Effective dose to a patient undergoing coronary angiography. AB - Detailed data relating to patient dose have been acquired for 18, randomly chosen, patients undergoing coronary angiographies. During the procedure, dose area product (DAP), kV and beam collimation were recorded for each projection. Total DAP, fluoroscopy time and number of cine frames were also noted. Patient effective doses were calculated using two different methods. Mean and median DAP values were 58 and 56 Gy.cm2 respectively, with a range between 26 and 125 Gy.cm2, screening time ranged from 3 to 37 min and cine frames from 1013 to 2344, depending on patient size, examination protocol and complexity of procedure. Our results correspond well with those obtained by other authors. Differences in the estimation of effective doses, due to the calculation method used and the approximation made, are discussed. PMID- 11487831 TI - Review of patient dosimetry in cardiology. AB - The objective of the study is to review doses to patients undergoing interventional cardiology procedures, doses that are known to be substantial due to prolonged fluoroscopy time and multiple radiographic exposures. High doses together with the fact that the use of radiation in cardiology is increasing, makes the assessment of patient dose and the establishment of reference dose levels necessary. Thus MED 97 can be better implemented in all EU member countries. The literature review has shown that data is limited and difficult to compare due to various dose indicators, different physical parameters of X ray equipment, various examination protocols and variations in patients' size and complexity of case. For these reasons, European countries participating in DIMOND II submitted data for percutaneous transluminal coronary angioplasty and coronary angiography. Reference dose levels were defined in terms of dose-area product, fluoroscopic time and number of frames. PMID- 11487832 TI - Stray radiation in the cardiac catheterisation laboratory. AB - Staff radiation risk is related to the radiation field in which individuals work. Traditional protective measures focus on reducing stochastic risk. However, deterministic injury to the operator's hands cannot always be ignored. The stray radiation field is almost totally attributable to scatter from the patient. Its relative intensity is greatest near the entry port of the useful beam into the patient. The entry port moves during the procedure as the operator selects various required projections. Therefore, the relative exposure rate at any particular location in the laboratory changes with the clinical projection. The absolute scatter intensity is also dependent on the size and strength of the useful beam. Operators may put their hands near or in the useful beam. Leaded surgical gloves provide some overall finger protection for scatter fields. However, because of automatic dose rate controls, these gloves often increase risk when the operator's hands are seen on the image monitor. PMID- 11487833 TI - Patient dose related to the complexity of interventional cardiology procedures. AB - In interventional cardiology (IC) the PTCA (percutaneous transluminal coronary angioplasty) procedure is the most frequent procedure with the highest dose to the patient. The procedure is usually performed by cardiologists having, in general, insufficient knowledge of radiation physics, radiation technology and radiation protection. The need for radiation protection is of paramount importance in this field of interventional radiology. Correlation between the complexity of PTCA procedure and irradiation parameters (fluoroscopy time, number of images and dose-area product--DAP) has been demonstrated. The presence of severe tortuosity and occlusion of > or = 3 months play a major role. Fluoroscopy time is better correlated with technical factors than DAP, which also includes the influence of patient size, collimation, operation modes, and X ray beam orientation. The number of acquired images is less correlated with the complexity of the PTCA procedure. A complexity index was conceived and procedures were divided into three groups, defined as: simple, medium and complex, which were significantly different in terms of patient dose. PMID- 11487834 TI - Medical exposures directive: implications for interventional radiology. AB - The new European Directive on Medical Exposures (97/43/EURATOM) specifically mentions 'high dose procedures' among the areas requiring special attention. This is universally interpreted as including the interventional procedures in radiology, cardiology and many of the special procedures involving fluoroscopy in these and related areas. This communication will focus on a definition of interventional procedures. It will examine the equipment, processes and patient and staff risks involved. From this examination it will be evident that new development is rapid in these areas and proceeding at a rate that is well ahead of both supporting research and regulatory framework. It will further note that the level of patient risk which is acceptable may be higher than in many other procedures. Finally it will note where the general provisions of the new Directive must be brought to bear on this new and developing field. PMID- 11487835 TI - Staff dosimetry protocols in interventional radiology. AB - Assessment of effective dose (E) for workers performing interventional radiology is particularly problematic due to the conditions of partial body exposure, so very few estimates of E are found in the literature. Two simple algorithms (the Rosenstein-Webster and the Niklason algorithms) are available that combine the readings of two dosemeters, one worn under the protective apron and one on the neck outside the apron, to estimate E for the range of imaging conditions typical of medical fluoroscopy. The algorithms are reviewed and their suitability for estimating E for personnel performing interventional radiology is analysed by comparison with the most recent experimental data. It can be concluded that the Niklason algorithm's estimates are in better agreement with the experimental assessments of E. PMID- 11487836 TI - Psychophysical aspects in medical illumination techniques. AB - The relation between image and visual perception of the human eye is an important point in digital imaging systems. Research aims should therefore pay attention to psychophysical aspects. Optimising of digital imaging systems can only be reached if the important final steps in the diagnostic process--visual perception and signal detection--are taken into account. PMID- 11487837 TI - Practical implementation of the medical exposure directive (97/43) in Luxembourg. AB - The medical exposure Directive of 1984 was updated in 1997 (EURATOM 97/43) and should be implemented into national legislation before May 2000. Need for harmonisation exists in Europe, as differences in the application of ionising radiation exist throughout European countries and social mobility of the population is increasing. The context of the justification/optimisation process as well as specific characteristics of national health legislation and Social Security legislation and their impact on radiation protection of the patient, are presented. Existing differences in the justification process throughout Europe are: (i) prescribing process of the X ray examination, training of the prescriber, (ii) auto-referral; (iii) economic situation in the member states. The optimisation process is mainly sensitive to: (i) 'culture' of radiation protection in each national medical community; (ii) training; (iii) technical and clinical audits; (iv) quality assurance. PMID- 11487838 TI - The quality criteria concept: an introduction and overview. AB - The Quality Criteria concept has been developed over the past decade in Europe and applied with success for conventional X ray examinations of adult and paediatric patients. This concept has recently been extended to computed tomography, and will also be available for digital radiography in the near future. The aim of the Quality Criteria for diagnostic images is to define a level of performance considered necessary to produce images of standard quality for a particular anatomical region and which could address any clinical indication. The image criteria include anatomical criteria, which relate to the visualisation or critical reproduction of anatomical features and also physical criteria measurable by objective means. The diagnostic reference doses introduced by ICRP 73 are an essential element of the Quality Criteria concept given for examinations on standard-sized patients. The Quality Criteria should provide a logical framework for radiation protection initiatives which links the desired or acceptable outcome, in terms of image quality, of a radiological examination, to the radiographic technique required to produce this outcome and the patient dose which should be achievable. PMID- 11487839 TI - Clinical aspects of quality criteria in digital radiography. AB - In clinical radiological practice two main questions regarding image quality have to be looked at: Which degree of image quality is needed? and: How can this image quality be objective? Image quality requirements depend on size, density and contrast of the objects of interest and on the clinical question which has to be answered. In comparison to conventional radiography, the digital radiographic techniques offer additional features concerning optimisation of image quality and dose, like a wider dynamic range, digital fluoroscopic techniques and post processing. Therefore it is necessary to define new quality criteria for these techniques. In the following, typical examples demonstrate the development of the 'Diagnostic Requirements for Digital Radiographic Procedures'. These examinations include projection radiographs with digital luminiscent radiography, fluoroscopic procedures with digital image intensifier radiography and angiographic procedures with digital subtraction angiography. The clinical question of a radiological examination also forms the background to optimisation strategies considering image quality and radiation dose. PMID- 11487840 TI - Methods and materials for the measurement of subjective and objective measurements of image quality. AB - The evaluation of medical imaging systems may be performed using both subjective (e.g. high contrast limiting resolution, and low contrast detail detectability), and objective/physical (e.g. characteristic curve, signal-to-noise ratio, modulation transfer function, and Wiener spectra) indices of image quality. For the purposes of Quality Assurance subjective measurements currently have a wider usage in the clinical environment due to the inherent simplicity of the measurements; however, more rigorous objective approaches are gaining acceptability, and represent a paradigm for possible future QA measurements. Both approaches have their advantages and disadvantages. There is currently wide debate as to the interpretation of these measurements. However, there appears to be a lack of standardisation among scientists in the various approaches to making these measurements, and especially so in the case of objective indices; furthermore, the measurement protocol used must have a bearing on the interpretation of subsequent results. The purpose of this paper is to (i) present the most fundamental indices in the characterisation of 'image quality', and (ii) recommend approaches to the measurement of these indices. PMID- 11487841 TI - Equipment requirements and specification for digital and interventional radiology. AB - During the past decade there has been a substantial growth in digital and interventional radiology. Equipment requirements and specification for digital (interventional) radiology are necessary to facilitate the purchase of proper installations for specific purposes. Inappropriate equipment might lead to increased dose to patients and staff, insufficient image quality and, for interventional radiology, to inefficient procedures and the potential for deterministic effects to occur. The equipment requirements and specifications are of various types. Requirements for dose displays and dose record keeping are dealt with in a separate contribution to this workshop. Detailed information is presented in this contribution on requirements and specifications in relation to ergonomic, dosimetric and image quality aspects. PMID- 11487842 TI - The quality assurance and constancy checking of fluoroscopy and fluorography systems. AB - X ray procedures which involve fluoroscopy often give significant radiation doses to patients. A valuable radiation protection procedure is the adoption of a quality assurance programme which will ensure the continual production of optimal quality images with the minimum necessary dose to the patient. Various publications exist which provide information on quality assurance (QA) and constancy checking of fluoroscopy and fluorography systems. These publications were reviewed and tests recommended for QA and constancy checking of fluoroscopy and fluorography systems are presented. It was found that not much information exists on QA and constancy checking for digital subtraction angiography and digital fluorography systems. More research is required in this field. Information on protocols used by various countries of the European Community for QA and constancy checking of fluoroscopy and fluorography systems was gathered. From this information it is apparent that there is a need for harmonisation within Europe. PMID- 11487843 TI - A critical overview of acceptance testing using various measured indices. AB - It is a requirement of the new European directive on health protection of individuals against the dangers of ionising radiation in relation to medical exposure (97/43/EURATOM), that acceptance testing is carried out on all radiological equipment before its first use for clinical purposes. This directive must be implemented into member state law. In accordance with this requirement, thirteen digital fluorography systems were acceptance tested by the Department of Medical Physics and Bioengineering, St James's Hospital, Dublin. The equipment tested was in six Irish hospitals and included nine systems intended for interventional use, two of which were dedicated cardiac catheterisation laboratories and two of which were dedicated vascular laboratories. Testing was performed in line with current published international guidelines and standards and involved the measurement and analysis of various indices including X ray tube and generator performance, operating dose levels, image quality and electrical, mechanical and radiation safety aspects. The results are presented and compared with international standards and recommendations. Problems were identified with all systems tested with approximately 70% of all systems demonstrating significant problems. This study emphasises the importance of acceptance testing radiological equipment. PMID- 11487844 TI - Review of acceptability criteria for X ray systems relevant for digital radiology. AB - The Medical Exposure Directive (97/43/EURATOM) states that competent authorities shall adopt specific criteria of acceptability for equipment in order to indicate when remedial action is necessary. To assist the competent authorities of the European Union Member states, the EC has prepared a document on criteria of acceptability for radiological (including radiotherapy) and nuclear medicine installations. The purpose of the document was to specify minimum standards of performance. Specific criteria for digital equipment are not given in the EC document. In the guidelines for quality control of equipment used in diagnostic radiology in the Netherlands limiting values as well as measurement methods are included. However, guidelines specific for digital imaging equipment are lacking. At present, Report 77 of the Institute of Physics and Engineering in Medicine is the most comprehensive document within Europe for quality control of diagnostic X ray imaging systems. It covers digital fluoroscopy systems and includes or refers to test methods. It contains test frequencies, levels of expertise required, priority of tests and remedial and suspension levels. In the present contribution the available criteria of acceptability for equipment relevant for digital radiology are summarised. PMID- 11487845 TI - Introduction to constancy check protocols in fluoroscopic systems. AB - Reference dose or guidance levels are a well established approach to the reduction of patient doses in diagnostic radiology. There are two main methods of determining reference doses, one involves patient dose measurements and the other phantom dosimetry. The latter approach lends itself to the development of constancy test protocols, which may be used as part of an acceptance testing programme or to compare the performance of different imaging systems. Various constancy test protocols and procedures have been proposed and these are reviewed. The constancy test protocols developed within the DIMOND concerted action will be described in detail. The advantages and disadvantages of the various methods and approaches are compared and contrasted. The complementary nature of constancy check protocols with patient dosimetry studies is discussed. PMID- 11487846 TI - Comparative reject analysis in conventional film-screen and digital storage phosphor radiography. AB - The purpose of this work was to gather information about the benefits in patient care caused by the introduction of digital radiography. In particular, the possibility of reducing the number of image repeats and thus unnecessary patient radiation was sought. Waste films of conventional radiography were collected--in digital radiography each image delete command at the post-processing workstation was documented. Rejected images were analysed retrospectively. The overall reject rate was 27.6% in the conventional and 2.3% in the digital department. While in the conventional department the main reason for rejection was 'exposure' and 'others' (i.e. problems related to film handling), the main reason in the digital environment was 'positioning'. Reject analysis yields representative data about the current performance of a radiology department. A marked reduction of repeated X rays and consequently reduced radiation exposure of the patient was clearly shown in this study comparing two differently working radiology departments. This is one of several benefits of digital radiography in patient care. PMID- 11487847 TI - A review of image quality and dose issues in digital fluorography and digital subtraction angiography. AB - This paper addresses image quality and dose issues in Digital Fluorography (DF) and Digital Subtraction Angiography (DSA), commencing with a brief review of methods and protocols for image quality assessment in DF and DSA. The relative scarcity of standards and test protocols for unsubtracted DF is highlighted. Pooled results from a large number of quality assurance tests are then used to illustrate trends in the choice of radiation dose per image currently employed in typical DF and DSA work. It is concluded that, although relatively high doses per image may be justified in terms of image quality improvement for DSA, there is little justification for the large range of exposures used in DF for nominally identical examinations. It is argued that the use of high doses per image in DF would not be expected to offer an advantage in terms of image signal-to-noise ratio. PMID- 11487848 TI - The assessment of dose and image quality of storage phosphor systems: an overview of the different tasks. AB - The assessment of the dose-image quality relation using storage phosphor systems requires a series of tasks. In the text, the main aspects are summarized and the differences with the measurements for X ray equipment using film-screen cassettes are indicated. Standards of testing are becoming available for quality control measurements of the equipment. The global analysis is, however, less standardised. Large scale trials concerning dose and image quality are missing. In particular, guidelines about the appropriate use of post processing parameters and image visualisation are scarce. In this regard, a newly proposed method to evaluate image quality as a function of post-processing parameters for particular types of clinical images is mentioned. It is concluded that a thorough analysis may be very time consuming, but if the properties of the digital detectors are exploited, many tasks can be automated. PMID- 11487849 TI - Optimisation of dose per image in digital imaging. AB - Three current digital radiography modalities are briefly described: digital fluorography, storage phosphor radiography and amorphous selenium radiography. For all of these modalities, the dose used to form an image can be varied considerably. Threshold contrast detail techniques were used to investigate the consequence of changing dose per image at the image receptor. At low doses, contrast resolution is limited by X ray quantum noise while system noise limits contrast resolution at high dose per image values. PMID- 11487850 TI - An assessment of methods for monitoring entrance surface dose in fluoroscopically guided interventional procedures. AB - In the light of a growing awareness of the risks of inducing skin injuries as a consequence of fluoroscopically guided interventional procedures (FGIPs), this paper reports measurements of ESDs made during the period August 1998 to June 1999 on 137 patients undergoing cardiac, neurological and general FGIPs in the South London area. In addition, different methods suitable for use in a routine monitoring programme of ESD are discussed. Although the need for additional data is acknowledged, the results reported reinforce the need for routine assessments to be made of ESDs in FGIPs. At present, the most reliable and accurate form of ESD measurement would seem to be arrays of TLDs. However, transducer based methods although likely to be less accurate, have considerable advantages in relation to a continuous monitoring programme. It is also suggested that there may be the potential locally for threshold dose area product (DAP) values to be set for specific procedures. These could be used to provide early warning of the potential for skin injuries. PMID- 11487851 TI - The value of digital personal dosemeters in angiography/interventional radiology: preliminary experience. AB - New interventional procedures tend to involve longer screening times than were hitherto used in radiology. A careful audit of technique and shielding facilities needs to be performed to ensure that patient and operator doses are optimised. This paper explores the use of digital dosemeters to evaluate operator dose. Equipment related parameters, e.g. screening time, dose-area-product (DAP) readings, were not found to be strongly correlated to operator dose. The real time display on the electronic dosemeter is a non-intrusive indicator of the efficacy of operator protection strategies. PMID- 11487852 TI - Deterministic effects in interventional radiology. AB - Since the development of interventional radiology, the number and complexity of procedures has increased significantly and continues to grow. Interventional radiology procedures offer substantial health care benefits. However, associated with the increasing complexity as well as a lack of quality control programmes and specific training in radiation protection, there is an increase in the occurrence of deterministic effects in both patients and staff. There is a growing literature of case reports describing deterministic effects. A review of some case reports and the response of various international organisations is presented. It is important that workers in interventional radiology are aware of the potential for deterministic effects and the dose threshold for their onset, so that patients can be followed up appropriately. PMID- 11487853 TI - Staff dosimetry in interventional cardiology. AB - In interventional cardiology (IC) staff can be exposed to high dose levels due to the long fluoroscopy procedures performed. Staff dosimetry can yield information on the optimisation level of radiation protection, which is influenced by the equipment performance, auxiliary protection devices, training in radiation protection and procedure complexity. Staff exposure data assessed in haemodynamic laboratories of four hospitals in Spain, Greece and Italy participating in the DIMOND concerted action are analysed and compared with data in the literature. PMID- 11487854 TI - [Implementation of new regulations in radiation protection in Germany]. PMID- 11487855 TI - Compensators for IMRT--an investigation in quality assurance. AB - Intensity modulated radiotherapy (IMRT) allows dose distributions which adequately consider organs at risk (OAR) and dose homogeneity to the target volume. This is practically reached by conforming the beam profiles to the shape of the planning target volume (PTV), by shaping the fluence with multileaf collimators (MLC) or compensators. Though compensator production is time consuming and seems less convenient than the use of MLC, compensators offer much easier quality assurance. In this study the effects of certain simplifications of compensator production were studied. Compensators were produced and ionization chamber measurements in a water phantom and film measurements in a solid phantom were performed to verify the compensators. The results of the measurements were compared to the fluence distributions given by the planning system. The measurements were meant to show how realistic the investigated simplifications were, and to reveal a suitable and reliable testing method for compensators. Monte-Carlo calculations employing the EGS 4 Code were further performed to support the measurements. PMID- 11487856 TI - [Evaluation of the electron density phantom CIRS Model 62]. AB - The properties of the electron density phantom CIRS M62 were studied. Physical and electron densities were calculated for all tissue substitutes, and compared to the values given in the data sheet and to the recommendations of the ICRU Report 44. The phantom was scanned in standard-CT and spiral-CT mode with different acquisition parameters. Especially manufactured tissue substitutes for dense bone were analysed for artifacts. Hounsfield values of all tissues substitutes were determined with the treatment planning system CadPlan. Calibration curves were calculated and compared to a stoichiometric calibration for real tissue. There was good agreement between the values calculated and the values given by the manufacturer. The tissue substitutes were only approximately tissue equivalent. The tissue substitutes for dense bone showed small artifacts. The calculated calibration curves were in good agreement with the stoichiometric calibration curve. PMID- 11487857 TI - [Constancy check of the radiation output of Linac: comparison with diodes and ionization chamber]. AB - For routine QA measurements of the radiation output of a linear accelerator, an ionization chamber is usually used, the type of which may depend on the radiation quality. In this study, a comparison of the standard ionization chamber and the central diodes of an array, was performed both used simultaneously for the constancy check of the radiation output. It was investigated to which degree the two detectors deviate from each other over a long period of time for different radiation qualities. The aim of the study was to show whether the radiation output can be checked using the diode array, which is usually used for measuring beam profiles, by averaging the values of the central diodes within a reasonable accuracy. The results showed a maximum deviation of the two detectors of 0.9 2.2%, whereas the mean deviation is 0.4-0.8%, both depending on the radiation quality. PMID- 11487858 TI - [Effect of ultrasound on membrane surface potentials]. AB - The electrostatic equilibrium on the surface of an ion-impermeable membrane was not influenced by ultrasound fields. Only after incorporation of an ion transporter did the ultrasound induce changes of the membrane surface potential. Because the ultrasound effect was completely reversible, measurements of the surface potential of a flat lipid bilayer membrane containing the calcium transporter calcimycin were performed, simultaneously to the ultrasound exposure. The ultrasound-induced volume flow, also called quartz wind, favored the mass transfer through the diffusion boundary close to the membrane, thereby leading to increased calcium concentrations in the immediate vicinity of the membrane. This, in turn, became manifest as a reduction of the negative surface charge density. PMID- 11487859 TI - [A testing device for quality assurance of 3D-ultrasound equipment]. AB - A test object for 3D-ultrasound (US) imaging equipment was constructed, that could be used for various purposes: For training of the correct technical setup procedures and practical handling of a 3D-US equipment, as well as for various quality assurance procedures. The size and volume of these structures were known precisely (+/- 0.1 mm resp. +/- 0.06 mL, respectively; smallest volume: 0.1 mL). The stability of the structures was in agreement with guideline IEC 854 and allowed to quantify exactly the imaging and measurement errors of size and volume measurements performed with commercial 3D-equipment. The test object is well suited to validate measured volume data, as well as to perform procedures of technical quality assurance in a reproducible manner. PMID- 11487860 TI - [Event-related functional magnetic resonance imaging of cerebral pain processing]. AB - Neurofunctional magnetic resonance imaging (fMRI) offers the possibility to map cerebral activity non-invasively. The development of event-related techniques during the past years allows to study brain processes with high spatial and temporal resolution. Based on these techniques, EPI- and FLASH sequences were developed in this study, to investigate cerebral processing of experimental thermal pain stimulation. Phasic and tonic stimulation paradigms were developed with an MR-compatible contact thermode. Functional mapping of pain-relevant areas was performed with these paradigms, as well as a specification of the temporal characteristics of the activation. Further, a randomized paradigm with several stimulus intensities could differentiate graded functional responses, dependent on stimulus intensity in specific "regions-of-interest". In this design, randomizing the stimulus order reduced habituation effects, while continuous subjective magnitude estimation of the stimuli kept attention of subjects maximal. PMID- 11487861 TI - [Pin-hole camera studies of the head scatter contribution and the photon source size of a linear accelerator]. AB - There are different opinions of the jaws scatter contribution to the photon fluence of medical linear accelerators. Investigations were performed at the Radiology Clinic of the University of Jena to give an experimental answer to this question. Since the study object was the linear accelerator LUE, now out of operation, the method of investigation is was the topic of this report. By use of a pin-hole camera and a "beam stopper", the focal and extrafocal radiation was projected upon the image plane of the camera and was measured with films or a Schuster detector array. The following relative contributions of the particular source regions to the total photon fluence in the field center were obtained: flattening filter (91.8 +/- 1)%, primary collimator (7.2 +/- 0.9)%, collimator jaws (1.0 +/- 0.3)%. These results are in good agreement with those of Ahnesjo. For this accelerator, the 37% width of the photon source region is (20 +/- 1) mm. PMID- 11487862 TI - [The Plexitom, a new quality assurance tool for the linear accelerator]. AB - Continuous depth dose measurements are generally performed in water phantoms. It is now possible to make continuous depth dose measurements in the solid PMMA phantom Plexitom, developed by our group and manufactured by PTW. The mechanical construction of this phantom allows the continuous movement of the ionisation chamber within the solid phantom. The Plexitom is light and very handy. It is very useful for quality assurance, especially to check the radiation quality of the linac. PMID- 11487864 TI - Problems of food allergen extracts. PMID- 11487863 TI - Assays for the determination of the biological activity of allergen extracts. PMID- 11487865 TI - Local routes of immunotherapy. PMID- 11487866 TI - The European Guidelines regulating allergen products and the Ph. Eur. Monograph on allergens: transformation into everyday practice. PMID- 11487867 TI - A clinical trial with purified natural major allergens from grass pollen. PMID- 11487868 TI - Vaccination strategies in young children at risk for allergy. AB - T-cell responses against foreign antigens/allergens are common findings very early in infancy and these responses are generally Th2-skewed. Normally, these responses deviate into low level Th1-type of immunity over the first years of life. In atopic children, however, the Th2-skewing persist, resulting in persistent IgE responses against allergens and the development of allergic diseases. In healthy children, tolerance induction by environmental and immunological factors seems to be related to an immune deviation towards a Th1 type of immune response. Hypothetically, immune deviation could be achieved in atopic children by vaccination with inhalant allergen and will conceivably prevent sensitization and development of respiratory allergy. This hypothesis is currently being tested in Sweden and Germany. PMID- 11487869 TI - Requirements for demonstrating clinical efficacy: clinical aspects. PMID- 11487870 TI - Requirements for demonstrating clinical efficacy: statistical aspects. AB - Usual statistical practice in inference on the comparison of two treatment effects is based on significance tests. In the positive case, it is unlikely that the observed difference is based only on random variation, and a decision is made that there is a real superiority. In the negative case, no conclusion on a missing difference is possible. In both cases, statistical tests fail to describe the magnitude of the underlying effect. Confidence intervals are more suitable for demonstrating clinical efficacy. Their advantages and applications are discussed. PMID- 11487871 TI - Recombinant allergens expressed in E. coli: benefits and drawbacks in the diagnosis of food allergies. PMID- 11487872 TI - Cloning and characterization of Ph1 p 1 in different expression systems. PMID- 11487873 TI - Expression of yellow jacket and wasp venom Ag5 allergens in bacteria and in yeast. AB - Antigen 5 (Ag5), of unknown biological function, is one of the major venom allergens of vespids and fire ants. We have compared the expression of Ag5 in bacteria and in yeast. Recombinant Ag5 from bacteria formed an insoluble intracellular product, which was not properly folded, but that produced in Pichia pastoris was secreted to the extracellular medium. Immunochemical characterizations showed the secreted Ag5 to have the native structure of the natural protein. This is of interest since the B cell epitopes of Ag5 are mainly of the discontinuous type. These studies were made with Ag5s from yellow jacket (Vespula vulgaris) and paper wasp (Polistes annularis), and with hybrid Ag5 molecules that contained partial sequences of these two species. In vitro allergenicity studies with sera from yellow jacket-sensitive patients showed that some of these hybrid molecules had a greatly reduced allergenicity but retained the immunogenicity of the natural allergen. This could be of importance for immunotherapy of this type of allergy. PMID- 11487874 TI - Biological activity of recombinant bee venom allergens expressed in baculovirus infected cells. PMID- 11487875 TI - Structural and biological demands on recombinant allergen related to their application. PMID- 11487876 TI - Skin testing with wild-type recombinant birch pollen allergens and hypoallergenic modified molecules. AB - Our aim was to compare the allergenic activity of wild type rBet v 1 with recombinant Bet v 1 derivatives (rBet v 1 fragments, dimer and trimer) with potentially reduced anaphylactic activity by skin testing in a French population. METHODS: Among the 36 birch pollen allergic patients included in the study, 29 were tested by skin prick testing and 30 by intradermal injections with purified monosubstances: rBet v 1 fragments (F1: aa1-74 and F2: aa75-160), Bet v 1 dimer and trimer. Intradermal tests were performed by the endpoint intradermal titration method. Tests were performed over a period of 6 months (before, during and after birch pollen season). RESULTS: All patients showed lower reactivity with the modified rBet v 1 allergens, both in skin prick- and intradermal tests. In 25 and 23 out of 29 patients the lowest concentration of fragment 1 and 2 respectively, resulting in a positive prick test was 100 fold higher than the lowest concentration of monomer resulting in a positive prick test. For dimer it was 100 fold or more in 25 out of 29 patients, and for trimer it was 100 fold or superior in 26 out of 29 patients. By intradermal testing, the end point concentration was 160 fold higher for trimer than for monomer in 24 patients and 40 fold higher in 5 patients. For the 2 fragments the end point concentration was 160 fold higher in 20 out of 22 patients. CONCLUSION: rBet v 1 fragments and trimer may represent candidate molecules for immunotherapy of birch pollen allergy with reduced risk of anaphylactic side effects. PMID- 11487877 TI - Allergen regulation in European countries. PMID- 11487878 TI - Large scale production and quality criteria of recombinant allergens. PMID- 11487879 TI - Registration of recombinant proteins (including allergens) in the EU. PMID- 11487880 TI - Expression and cloning of recombinant indoor allergens. PMID- 11487881 TI - Differential regulation of allergen-specific antibodies in allergy and specific immunotherapy. AB - Allergen-specific immunotherapy (SIT) aims to specifically skew an allergic response into a normal immune reaction against an allergen. The response to bee venom (BV) provides an especially suited model to study the immunological mechanisms of SIT in human. The BV-phospholipase A2 (PLA) represents the major antigen/allergen of BV. In SIT of BV allergy both whole BV and T cell epitope peptides of PLA were successfully applied. It appeared that the induction of specific anergy in peripheral T cells and reactivation of the T cells by microenvironmental cytokines represent the basic key steps in the immunological mechanism of SIT. The proliferative and cytokine responses by specific T cells were significantly suppressed simultaneously with an increase in IL-10 after 7 days. The anergic state was fully established after 4 weeks. Neutralization of IL 10 in PBMC by a specific antibody reconstituted the original proliferative and cytokine responses. Intracytoplasmatic cytokine staining revealed that IL-10 was initially produced by activated allergen-specific T cells. IL-10-producing B cells and monocytes were involved at a later stage of SIT and in maintenance of the anergy. The addition of IL-10 to stimulated PBMC or purified B cells inhibited IgE synthesis and enhanced the IgG4 antibody formation. Thus, SIT generates IL-10, which in turn induces specific anergy by autokrine interaction in T cells and counter-regulates IgE and IgG4 production. Particular cytokines from the tissue microenvironment reactivate the T cells to produce distinct Th1 or Th2 cytokine patterns respectively and by this way direct SIT towards successful or unsuccessful treatment. High amounts of allergen administered in SIT preferentially generate Th1 cytokines in T cells and IgG4 antibodies in memory B cells. Further investigations demonstrated that suppression of T cells by IL-10 is an active process, which depends on the expression and participation of CD28. PMID- 11487882 TI - Anti-IgE (rhuMAb-E25) therapy. PMID- 11487883 TI - Formulation and delivery of therapeutic allergy vaccines. PMID- 11487885 TI - Immunomodulation and immunotherapy using liposome entrapped allergens. PMID- 11487886 TI - Adjuvants for specific immunotherapy past, present, future. PMID- 11487894 TI - Status of allergen standardization in India. PMID- 11487889 TI - DNA based immunotherapeutics for allergy. PMID- 11487895 TI - Statistical considerations in the establishment of release criteria for allergen vaccines in the USA. PMID- 11487896 TI - Formulation of therapeutic allergen mixtures problems associated with the number, proportion, and enzymatic activities of allergens. PMID- 11487897 TI - Named patient preparations: interest for immunotherapy and possible regulation. PMID- 11487898 TI - Named patient products: critical aspects. PMID- 11487899 TI - Adaptation of registration dossiers to the requirements of the Directive 89/342/EEC (so-called "updating"). PMID- 11487900 TI - Quality improvements of allergenic source material. PMID- 11487901 TI - A new start for allergen references and standardization based on purified (natural or recombinant) allergens and monoclonal and monospecific polyclonal antibodies. PMID- 11487902 TI - Standardization in quality control of modified end-products. PMID- 11487903 TI - Gastric acid secretion in Helicobacter pylori positive and negative dyspeptic Nigerians. AB - The pathological role of Helicobacter pylori is largely unproven in our region of high incidence of infection but very low incidence of serious gastroduodenal lesions. The aim of this study was to investigate the effect of H. pylori infection on gastric acid secretion. One week after gastroduodenoscopy, basal and pentagastrin (8 micrograms/kg) stimulated gastric acid secretion were measured in 39 dyspeptic Nigerians. H. pylori status was determined using urease test, culture, histology and serology, while gastritis was assessed using the Sydney system criteria. The median maximal acid output (MAO) and peak acid output (PAO) in mmol/h were significantly higher in H. pylori positive (29.3, range 7.4-81.6 and 34.4, range 7.6-144.0) than in H. pylori negative (16.6, range 4.2-44.1 and 22.4, range 5.6-48.6) patients, p = 0.019 and p = 0.029, respectively. Stimulated gastric acid secretion was significantly higher in patients with duodenal ulcer (n = 8) than in H. pylori negative (n = 11) patients, but was similar in non ulcer dyspeptics (n = 20) and H. pylori negative patients. The median basal acid output was not significantly different between the groups of patients. Our patients (median age 32 years) had normal mucosa (12.1%), pangastritis with corpus predominance (12.1%), antrum-only gastritis (24.3%) and pangastritis with antral predominance (51.5%). In the subset of H. pylori positive patients (n = 28, 71.8%), there were no significant correlations between grade of antral chronic inflammation, gastritis index score, anti-H. pylori IgG titre and gastric acid secretion, p > 0.05. H. pylori infection increases MAO and PAO in our relatively young patients with antral predominant chronic gastritis. PMID- 11487904 TI - A 10-year review of venous thrombo-embolism in surgical patients seen in Jos, Nigeria. AB - A retrospective study was carried out on 42 clinically diagnosed cases of venous thrombo-embolism (VTE) seen over a 10 year period. The mean age at presentation was 41 years with a sex ratio M:F of 2.5:1. The three commonest primary surgical conditions in patients with VTE were spinal trauma (16 percent), benign prostatic hypertrophy (12 percent) and head injury (10 percent). Prostatectomy was performed in 18 percent, ventral hernia repair in 12 percent and splenectomy in 9 percent. Sudden severe dyspnoea was the clinical presentation in 60 percent of patients. Central chest pain occurred in 43 percent, loss of consciousness (38 percent), haemoptysis (7 percent), mental confusion (19 percent) and lower limb swelling (14 percent). Fifty-two percent of patients presented within 10-15 days. Oral warfarin and intravenous heparin were employed in management in 45 percent. Hospital mortality from massive pulmonary embolism was 64 percent while post thrombotic syndrome occurred in 5 percent of patients. PMID- 11487905 TI - Effect of HIV infection on the clinical spectrum of leprosy in Maiduguri. AB - The clinical features associated with different classes of leprosy patients co infected with HIV in Maiduguri was studied and the classification of leprosy was done clinically and bacteriologically using Ridley-Jopling classification and bacteriological index respectively. The cases were classified as paucibacillary (Tuberculoid--TT and Borderline Tuberculoid--BTT) and multibacillary (Borderline Borderline--BB, Borderline Lepromatous BL and Lepromatous Leprosy--LL) leprosy. Eleven (10.5%) of 105 leprosy cases were HIV-seropositive comprising of 7 males and 4 females. Age range was 15 and 62 years. Among the HIV seropositive patients, those with paucibacillary (PB) leprosy were 6 (TT-1, BT-5) while multibacillary (MB) leprosy 5 (BB-1, BL-2, LL-2). The predominant clinical features were clawing of fingers (64%), ulcerations (64%), hand muscle atrophy (55%) and clawing of toes (45%). Some clinical features of paucibacillary leprosy such as sensory and hair losses (as is also seen in HIV negative patients) occurred in increased frequency in HIV positive patients belonging to the multibacillary class. The HIV infected leprosy patients are more likely to manifest advanced stages of the disease than the HIV seronegative patients. PMID- 11487906 TI - HIV infection: risk of occupational exposure in a chemical pathology laboratory in Nigeria. AB - The prevalence of HIV/AIDS is rising globally, the worst affected area being Sub Saharan African. In order to assess the risk of occupational exposure to HIV/AIDS in laboratory workers in Nigeria, we screened 210 consecutive serum sample sent to the Chemical Pathology Laboratory of the University of Port Harcourt Teaching Hospital, Port Harcourt, over a two-week period. 17 (8.1%) of all the sample screened tested positive. The Sero-positivity rate was 11.3% for adults and 2.6% for children. The implications of these findings with regards to the risk of occupational exposure of laboratory and health worker is discussed. The need for more attention to be paid to laboratory safety regulation in sub-saharan Africa in highlighted. PMID- 11487907 TI - A comparative study of induction and recovery characteristics of propofol and midazolam. AB - Propofol and midazolam were compared in 40 adult patients in A.S.A. 1 or 2 presenting for short surgical procedures (< 70 minutes) with respect to induction time, pain on injection, apnoea, heart rates, blood pressure, oxygen saturation, time to eye opening on command. The first group was induced with midazolam (0.15 0.20 mg/kg) while the second was induced with propofol (2-2.5 mg/kg) intravenously. In all other respects except for the surgery that patients had the same treatment. The mean induction time was 55.25 + 26.66 and 69.75 + 24.72 for propofol and midazolam groups respectively. In the midazolam group apnoea occurred in 10% of patients compared to 80% of patients in the propofol group. Local reaction (phlebitis) was absent in the midazolam compared with 20% incident rate observed in the midazolam group. Propofol lowered blood pressure more than midazolam after three minutes of induction at a statistically significant level (P < 0.05). Recovery was significantly more rapid following propofol (P < 0.05). PMID- 11487908 TI - Features of childhood hepatic failure in Calabar, Nigeria. AB - The features in 21 patients with childhood hepatic failure studies retrospectively over a seven year period at the University of Calabar Teaching Hospital, Calabar, Nigeria are presented. Of the 21 patients, 71.4 per cent were aged three years nd below. Fulminant hepatic failure occurred in 81.0 per cent of the patients while in 19.0 per cent, the failure resulted from chronic liver disease. Hepatitis B infection alone or in association with other factors was the major cause of the condition, occurring in 76.2 per cent of the cases. The main complications were severe anaemia (23.8%), septicaemia (23.8%) and pneumonia (19.0%), renal failure (9.5%). With only one survival, the case fatality rate was 95.2 per cent. For prevention of the condition in Nigeria, universal mandatory screening of blood and blood products for hepatitis B markers before transfusion and the integration of hepatitis B vaccination into the National Expanded Programme on Immunization are strongly recommended. PMID- 11487909 TI - Aspirin-induced duodenal perforation in a Nigerian with sickle cell disease: a case report. AB - A case of duodenal perforation associated with aspirin ingestion in a 21-year old male Nigerian with sickle cell anaemia is reported. He presented with a sudden onset of epigastric pain which later spread to involve other parts of the abdomen. He had previously used aspirin at a dose of 1800 mg daily for two weeks to treat bone pain. Abdominal ultrasonography and X-ray showed fluid collection in the pelvis and elevation of the diaphragm. At exploratory laparotomy, the perforation found in the first part of the duodenum anteriorly was repaired. He was discharged 11 days post-surgery. In view of the frequent usage of salicylates and non-steroidal anti-inflammatory drugs to treat painful crises in sickle cell disease, we suggest careful monitoring of patients on such drugs and those with dyspeptic symptoms must be fully investigated including the use of endoscopy, to prevent fatal outcome. PMID- 11487910 TI - Right lobar pneumonia complicated by sub-phrenic abscess in a child. AB - A 14-year old boy presented with chest and abdominal pain and fever for one week. He had been treated with several antibiotics at home and in a peripheral hospital for respiratory infection. Physical examination showed features of right lobar pneumonia and peritonitis. Chest radiograph showed consolidation in the right lower lung field and abdominal ultrasonography showed a subphrenic collection. At exploratory laparotomy, a right subphrenic abscess and general peritonitis without an intra-abdominal focus were found. The abscess was drained and broad spectrum antibiotics given. Death, however, occurred from overwhelming infection. Subphrenic abscess complicating pneumonia is unusual but can be the cause of poor response to treatment. The diagnosis should be excluded in a child with pneumonia and persisting abdominal symptoms. Prompt treatment is necessary to avoid morbidity and mortality. PMID- 11487911 TI - Arrhythmias: appraisal of symptoms by haemodynamic concepts. AB - Arryhthmias can afflict any heart, even in the absence of disease. Given that Cardiac Output (CO) is a product of heart rate and stroke volume, the end-result of brady- and tachy-arrhythmias is the same: low CO with symptoms of low-output state. In bradyarrhythmias, myocardial inotropy operates maximally but the product of stroke volume and slow heart rate yields a low CO. In tacharryhthmias, myocardial inotropy is redundant because the product of low stroke volume arising from low Diastolic Filling Time (DFT) and heart rate also yields a low CO. The hallmark of extrasystole is the duplex of symptoms: dizziness followed by palpitation. PMID- 11487912 TI - Central venous catheters for haemodialysis: a review. AB - Indications for using central venous catheters (CVC) for haemodialysis include patients with: exhausted vascular access sites, no suitable vessels, failed peritoneal dialysis or short life expectancy. Catheter design and technology have changed in recent years to address the perennial problems of poor flow and infection. Permanent CVC offer a real alternative to arteriovenous access as 11 22% of long-term haemodialysis patients use CVC as their permanent vascular access. Recent advances in catheter use including measures to avoid insertion complications are highlighted. CVC provide mean flow rates of 274-525 ml/min with a one year cumulative catheter survival of 47-93%. Tunnelled CVC provide a safe and effective long-term access for haemodialysis and are particularly suitable for use in developing countries. They are cheaper to maintain and remove the temptation to re-use disposable dialysis needles. PMID- 11487913 TI - Office romances. PMID- 11487914 TI - Reflections of dentists in midpractice. PMID- 11487916 TI - Sensitive teeth. PMID- 11487915 TI - Professional money management: your financial health prescription. PMID- 11487917 TI - A woman with a mission. PMID- 11487918 TI - Successful treatment planning. Part 1: Communicating. PMID- 11487919 TI - The plight of patient rights. PMID- 11487920 TI - A new species of arboreal pitviper from the Atlantic versant of northern Central America. AB - A new species of green, prehensile-tailed pitviper of the genus Bothriechis is described from the Atlantic slopes of eastern Guatemala and western Honduras. This species appears to be most closely related to B. bicolor of the Pacific versant of Chiapas (Mexico) and Guatemala. Several other species of Bothriechis occur on the Atlantic versant of northern Central America, including two montane species, B. aurifer and B. marchi but, with one possible exception, these are not known to be sympatric with the new species and occur in different mountain ranges. The widespread B. schlegelii occurs up to at least 900 m on the Sierra de Caral, where the lowest elevation recorded for the new species is 885 m. PMID- 11487921 TI - [Temporal distribution of diurnal rapacious birds in "Playon de Mismaloya" Reservation, Jalisco, Mexico]. AB - Diurnal raptors were counted monthly in the "Playon de Mismaloya", reserve, Jalisco, Mexico, from November 1997 to October 1998. We identified 11 species; eight of them migrants, one resident and two resident with migrant populations. The highest increase in total number of individuals was from December to March, influenced mainly by the presence of Caracara polancus (Crested Caracara), Falco sparverius (American Kestrel) and Pandion haliaetus (Osprey). The two former species were observed frequently in open areas, where they could detect prey more easily, while ospreys were found only in estuaries, lagoons and beach, where food was available. PMID- 11487922 TI - Coexistence of reef organisms in the Abrolhos Archipelago, Brazil. AB - The first study on coexistence of reef benthic organisms in Brazilian coral reefs was done in three localities of the Abrolhos Archipelago. Organisms were recorded in concentric circle samples (10 and 20 cm in diameter) randomly laid on transects. Type and frequency of "coexistence events" between pairs of organisms were determined. Most frequent organisms (massive and branched coralline algae, Favia gravida, and Agaricia agaricites) also had many significant positive coexistence events. These results might be related to the abundances of these organisms. The most frequent coral (Siderastrea stellata), however, exhibited only a few significant coexistence events (9% of 32 tests). Since the great majority of events were positive, and since there was high variation in the species/groups involved in significant events in different localities, benthic communities of Abrolhos Archipelago may well be structured primarily by abiotic rather than biotic factors. PMID- 11487923 TI - [Asteroidea, Echinoidea y Holothuroidea in shallow bottoms of Bahia de Loreto, Baja California Sur, Mexico]. AB - To evaluate echinoderm distribution, abundance and density a double 50 m transect, with a side observation range of 2.5 m was used at each of 11 stations (bimonthly samplings, Aug. 1997-Febr. 1998). In Bahia de Loreto the Phylum Echinodermata consists of 26 taxa. The greatest mean abundance by transect and mean densities in order of importance for Echinoidea were: Echinometra vanbrunti (94.1 +/- 52.9 ind, 0.25 ind/m2), Centrostephanus coronatus (38.15 +/- 9.15 ind, 0.06 ind/m2), Tripneustes depressus (28.68 +/- 6.86 ind, 0.039 ind/m2), Eucidaris thouarsii (10.66 +/- 3.37 ind, 0.025 ind/m2) and Diadema mexicanum (11.75 +/- 4.92 ind, 0.023 ind/m2); for Asteroidea were: Phataria unifascialis, Mithrodia bradleyi and Acanthaster planci. Asteroidea was the dominant class with 12 species, followed by Echinoidea with ten and Holothuroidea was represented by four species. PMID- 11487924 TI - [Helminthofauna of Opisthonema liberate and Harengula thrissina (Osteichthyes: Clupeidae) from Chamela Bay, Jalisco, Mexico]. AB - We collected helminths from the "sardines" Harengula thrissina (N = 61) and Opisthonema libertate (n = 43), from Chamela Bay, Jalisco State, Mexico (12 and six species, respectively). The nematode Pseudoterranova sp., reached the highest values of prevalence and mean abundance in O. libertate (11.6% and 0.13 worms per host, respectively), while in H. thrissina the digenean Parahemiurus merus reached a prevalence of 49.1% and a mean abundance of 1.40 worms per host. Low similarity values (qualitative and quantitative) between helminths of both host species is a result of their opportunistic feeding habits (100% of the parasitic species in H. thrissina and 66% of those from O. libertate infect their host from prey) and differential exposure to helminth larvae. PMID- 11487925 TI - The vertical distribution and abundance of gastropods and bivalves from rocky beaches of Cuastecomate Bay, Jalisco, Mexico. AB - The vertical distribution and abundance of conspicuous gastropod and bivalve species were studied at five rocky beaches in Cuastecomate Bay, Jalisco. Sampling was done from September, 1993 through March, 1994 with 0.75 m2 quadrants placed along replicate transect lines (10 m long) in the supralittoral and mesolittoral (upper, middle and lower intertidal) zones. A total of 6,643 mollusks were collected. Gastropods dominated the samples (6,272 individuals, 44 species); the bivalves were less abundant and diverse (371 individuals, five species). Seventeen species comprised 89.8% of all individuals collected. The gastropods Nodilittorina aspera and Nerita scabricosta were the most abundant with 637.8 and 71.43 individuals/m2, respectively. The most abundant bivalves were Brachidontes adamsianus and Chloromytilus palliopunctatus with 60.7 and 61.3 individuals/m2 respectively. The abundance of gastropods decreased from the supralittoral to the lower tidal zones while the number of species increased in the same direction. The number of species of bivalves also increased from the supralittoral to the lower intertidal zone; the abundance of individuals was higher at the middle intertidal zone. Affinities between groups of species among sampling stations were identified by computing Pearson's correlation coefficient using abundance values (ind./m2) and Jaccard's dissimilarity index using species presence or absence in the lower intertidal zone. Affinity among stations was not dependent upon their vicinity but on the high dominance of few species, the occurrence of many secondary species and beach characteristics. PMID- 11487926 TI - [Composition, distribution and abundance of gastropod larvae in the South of Quintana Roo,Mexico and north of Belice]. AB - To know the composition, abundance and distribution of gastropod larvae, monthly samplings were carried out in the south of Quintana Roo, Mexico and north of Belize, from April to December, 1996. Collections were made in six sites at Chinchorro Bank, four in the South Coast and six at Hol-Chan, Belize, between the 10 and 20 hrs. At each station 2.5 m3 of seawater were pumped through a 202 microns mesh; 27 species were identified. The most abundant species were: South Coast, Rissoina sp. 1., Limacina sp. 1 and Natica sp. 1, Chinchorro Bank, Limacina sp. 1, Creseis acicula, Cerithiopsis hero and Rissoina sp. 1 and Hol Chan, Limacina sp. 2, Alaba incerta and Rissoina sp. 1. The highest abundance was in rainy season. Apparently the presence of winds, coastal currents and food availability, control the distribution and abundance of larvae. PMID- 11487927 TI - [Abundance and growth of Pinna carnea (Mytiloida: Pinnacea) juveniles in suspended-frame culture]. AB - We quantified the abundance of juvenile Pinna carnea at Turpialito, Golfo de Cariaco, Venezuela, using collectors suspended at 8 m depth for 16 months and then evaluated their subsequent growth in suspended enclosures over 14 months in relation to concurrent measures of temperature and planktontonic food availability. All body components showed a rapid increase during the first four five months. There was a decrease in somatic tissue mass and great variability in growth coincident at first spawning, suggesting a negative effect of gonadal production on somatic growth. A multiple regression analysis indicated a strong relationship between tissue growth and food availability as measured by organic sestonic matter. Although we determinated an adequated spat abundance for culture activities (80 spat per collector-30 x 60 cm), spat collection was limited, mainly from February through April. In addition to high variability in growth and negative influence of environmental factors during unfavorable periods, the muscles production required in 14 months, suggesting that P. carnea is not appropriate for commercial production of muscle. However, the high weight of all edible tissues mass attained (21 g) and the high survival rate in all experimentation (> 96%) suggests that P. carnea is fit for aquaculture. PMID- 11487928 TI - [Density, growth and survival of Lyropecten (Nodipecten) nodosus (Pteroida: Pectinidae) juveniles in suspended-frame culture in Golfo de Cariaco, Venezuela]. AB - The effect of culture density on growth and survival of juveniles was studied in the scallop Lyropecten nodosus with intermediate hanging cultures in Golfo de Cariaco, Sucre state, Venezuela, from December 1993 to July 1994. Six spat densities (7, 15, 30, 60, 125 and 250 scallops/nets) were evaluated. The initial shell height and dry mass tissue were 9.4 +/- 0.18 mm and 0.007 +/- 0.0026 g, respectively. The specimens were placed into pearl nets and suspended in a long line at 8 m depth. Growth patterns of somatic tissue were similar, but with different magnitudes and inverse proportionality to culture density. No effect of density was observed on survival. These results suggest that intraspecific competition of cultivated scallops cause stress, which affects growth but not survival. We suggest as strategy for the management of culture density, successive density reductions, to obtain in a six months culture (intermediate culture) specimens of 50 mm in shell height in order to start the final culture period. PMID- 11487929 TI - [Serpulids (Polychaeta: Serpulidae) from the Northwestern Caribbean with keys to the Grand Caribbean region: Salmacina, Ficopomatus, Pomatoceros, Pomatostegus, Protula, Pseudovermilia, Spirobranchus and Vermiliopsis]. AB - From the revision of more than 1,250 serpulids, 15 species from the Grand Caribbean Region were identified and characterized. Thirteen species were collected along the shores of the Yucatan Peninsula and eight were found in other localities in the Gulf of Mexico, seven others are from Cuba and comments on type specimens of two species are also included. Three morphometric analyses were made to evaluate some characters. The first on Pomatostegus stellatus (Abildgaard), and related species and subspecies: P. brachysoma Schmarda, P. macrosoma Schmarda, P. s. fruticosa Morch, P. s. pentapoma Morch and P. s. tetrapoma Morch, indicated that they are conspecific. Another one on Spirobranchus de Blainville species: S. dendropoma Morch, S. giganteus (Pallas), S. polycerus (Schmarda) and S. polycerus augeneri ten Hove; indicated that S. giganteus differs from the other three taxa. The third analysis was made on Vermiliopsis annulata (Schmarda) sensu lato; it allowed the recognition of three different forms. Several incomplete specimens might belong to an undescribed genus. A key for identification of all the species recorded in the Grand Caribbean Region is also included. PMID- 11487930 TI - [Serpulids (Polychaeta: Serpulidae) from the Northwestern Caribbean: Hydroides and Serpula]. AB - From the study of more than 450 specimens of Hydroides and Serpula, 12 species from the Grand Caribbean Region were identified and characterized. Eight species were collected along the shores of the Yucatan Peninsula and five were found in other localities in the Gulf of Mexico, seven others are from Cuba and comments on type specimens of five species are also included. One morphometric analysis made on Hydroides mucronatus Rioja and Hydroides cf. mucronatus, indicated several differences among them. Comments on all species are also included. PMID- 11487931 TI - Production of the rotifer Brachionus plicatilis (Ploimida: Brachionidae) in a Brazilian coastal lagoon. AB - Secondary production, density and biomass fluctuations, and some reproductive aspects of the rotifer Brachionus plicatilis were studied weekly for two years in a Brazilian coastal lagoon. Food (and secondarily temperature) seemed to be the main factor involved in the summer increments. In the first summer, a mass mortality of fish caused changes in the phytoplankton, which switched from the dominance of Cyanophyceae to Chlorophyceae and other groups. For two months, production, density and biomass were relatively high until the recovery of the Cyanophyceae dominance. In the second summer, peaks of shorter duration and higher values of production, in comparison to the first one, were found. The contribution of resting eggs hatching, in the first summer, and the increment in parthenogenetic reproduction, in the second summer, would account for the population increases observed. Males and mictic females occurred when densities were high. PMID- 11487932 TI - Does damming of the Colorado River affect the nursery area of blue shrimp Litopenaeus stylirostris (Decapoda: Penaeidae) in the Upper Gulf of California? AB - After damming the Colorado River the freshwater flow was reduced to 1% of its virgin flow to the Upper Gulf of California (UGC). The ecological effects need to be properly documented. The UGC is the nursery area for Litopenaeus stylirostris, the most profitable fishery in the zone. In order to know the relative abundance of L. stylirostris postlarval stage we conducted a sampled survey every 14 days in 1993, 1994 and 1997, plus an intensive sampling during a complete tide cycle in July 1995 and 1996. We did 10 min trawls each hour during the flood tide. Relative abundance of postlarvae was higher (p < 0.05) in those years when freshwater flow reached the UGC. PMID- 11487933 TI - [Composition by size, age and growth of Litopenaeus vannamei (Natantia: Penaeidae), in Mar Neutro lagoon, Oaxaca-Chiapas, Mexico]. AB - The commercial catches of white shrimp Litopenaeus vannamei were sampled (May through November 1996) in the Mar Muerto lagoon located between Oaxaca and Chiapas states, Mexico. To measure it's growth rate at the innermost portions of this system, 16,378 length and weight records were obtained. Length fluctuated between 11 and 145 mm, and weights between 0.1 and 26.3 g. The minimum mean size for recruitment to the exploited phase is 45 mm. Organisms leave the lagunar system for recruitment to the marine zone 83-120 mm/length, at an age of 1.5-4.5 months. Analysis of modal progression detected 13 generations inside the lagoon. The growth rates ranged between 0.13 and 1.27 mm/day, with a mean of 0.64 mm/day. The asintotic lengths fluctuated between 105 and 159 mm with a mean of 136 mm, while the asintotic weight fluctuated between 8 and 23 gr with a mean of 15 gr. The parameters of the von Bertalanfy growth model are: L infinity = 136 mm, P infinity = 15 gr, k = 0.0161 mm and to = 5 mm. Both, L infinity and P infinity doesn't represent, at least in this case, the asintotics sizes and weights that the species reach inside the system, but should be interpret like the means sizes and weight that should have the organisms at the time to leave it. PMID- 11487934 TI - [Reef fishes community structure in 4 atolls of the San Andres - Providencia Archipelago (Southwestern Caribbean)]. AB - In 1994 and 1995, 131 visual censuses of reef fishes were made using the stationary sampling method in Courtown, Albuquerque, Serrana and Roncador, four atolls of the Archipelago of San Andres and Old Providence in the Southwestern Caribbean. Fish species and their abundances were recorded in four geomorphologic zones: lagoon, windward barrier reef, windward terrace and forereef terrace. A total of 98 species were censused; the most abundant were Chromis cyanea (14%), Clepticus parra (14%) and Stegastes partitus (10%). The most abundant families were Pomacentridae (37%), Labridae (28%) and Scaridae (10%). Analysis of similarities showed that differences between zones were greater than differences between atolls, but lagoon and forereef terrace were not significantly different. Cluster and ordination analysis confirmed these results; in addition, the ordination analysis placed the groups according to depth and wave-exposure gradients, suggesting that these two physical variables were responsibles for the clustering. Differences in equitability and species richness appear also due to these variables. Inverse analysis showed in each group few characteristic species, then the differences between zones were due specially to dominance of some species. The dominant trophic categories were planktivorous and herbivorous that were significantly different between zones. In shallow zones (shallow lagoonal patch reefs) and high wave-exposed zones (winward barrier reef) dominated herbivorous fishes, while in deeper zones (terraces and deep lagoonal patch reefs) planktivorous were more abundant. PMID- 11487935 TI - [Index of the Venezuelan marine microflora: diatoms, dinoflagellates and cocolithophorids]. AB - The marine phytoplankton of Venezuela has been studied on a regular basis since the mid 20th century. However, a species checklist that can be used as a framework for taxonomic studies is lacking. In this paper, an index of the marine microflora of Venezuela is presented for the first time. The index includes only those diatoms (89 centric and 186 pennate species), dinoflagellates (eight naked and 154 thecate species) and coccolithophores (24 species) for which formal diagnosis and illustrations (drawings and/or photographs) have been reported in the scientific literature (journals and/or first degree or master's theses). It is ordered alphabetically according to class, order, family, and species. It includes the author (s) of the taxa. PMID- 11487936 TI - The effects of electric impulse on growth of Rhizophora mucronata seedlings (Rhizophorales: Rhizophoraceae). AB - Exposure of mangrove seedlings to electric current was studied at an intensity of 10, 20 and 30 mA to 0.5, 1, 3, 6, 12 and 24 h in different combinations, either to plumule or radicle or both parts of the propagules of Rhizophora mucronata Poir (sample size = 30). The electric current-treated propagules and untreated control were cultured hydroponically in sea water for 75 days, and root and shoot growth parameters measured. The root elongation was significantly influenced by duration of exposure, site of propagule treated and combined effects of current intensity x site of propagule treated. Leaf number and area were significantly influenced by intensity of electric current and duration of exposure; whereas shoot elongation was affected by the duration of exposure. The root number was significantly affected by duration of exposure. In general, the electric impulse improved the growth characteristics of shoot and root. PMID- 11487937 TI - An upgraded method to relocate marked shoots of the seagrass Zostera marina. AB - This paper presents a new method for the recovery of marked seagrass blades. The introduction of a plastic belt surrounding the marked shoot at a sediment level provided a relocation arrangement which was unloosed by drag forces or grazing. The relocation method was tested on Zostera marina L. It proved to have the advantage of increasing dramatically the number of marked shoots recovered up to 100% while reducing the cost of the procedure to a minimum. An allometric model indicated that the introduced relocation method has no impact on the development of the plant. PMID- 11487938 TI - [Macroscopic fungi from Sierra de Quila, Jalisco, Mexico: diversity and fungal similarity]. AB - Diversity and similarity of macrofungi of Sierra de Quila, Jalisco, Mexico were analized in three different kinds of vegetation. Fungal diversity in the area is high. The pine-oak and cloud forests, were more diverse in their community structure than the oak forest. Similarity among the three kinds of vegetation was low, there are few species share among them; pine-oak and cloud forests show higher affinity than oak forests. This pattern of similarity is a general condition for others regions with environmental conditions similar to Sierra de Quila. PMID- 11487939 TI - [Description, distribution, anatomy, chemical composition and uses of Mimosa tenuiflora(Fabaceae-Mimosoideae) in Mexico]. AB - Because of some catastrophic events which occurred in Mexico during the 1980 decade, the utilization of "tepescohuite" bark against skin wounds and burns was popularized. The media manipulated the lack of available information about its medical properties and gave erroneous information to the society propagating a lot of myths. Therefore, the aim of this paper is to determine its taxonomic identity and to study the distribution, bark and wood anatomy of this species, and to determine its actual and historic uses, and the compilation of the information about bark pharmacology and toxicity. Its taxonomic identity is established as Mimosa tenuiflora (Willd.) Poir. (Fabaceae-Mimosoideae). It blooms and fructifies from November to June, occurring in Mexico (the states of Oaxaca and Chiapas), Guatemala, Honduras, El Salvador, Nicaragua, Panama, Colombia, Venezuela and Brazil, at altitudes of 0-1110 (-1520) m. In Mexico, it is found in dry forests, thorny thickets, Pinus and Pinus-Quercus forests, and in M. tenuiflora pure thickets, along roads and in resting or abandoned culture lands. This species has an aggregate distribution in the forests and a uniform one in the thickets. It presents a mean density of 9 individuals per m2, with 0.45 of frequency per quadrat and 1.69 m2 of mean coverture, and it has a wide range of tolerance to climatic and edaphic factors, confirming their invasive character. Regionally, the wood is used as fuel and fence construction, and against skin wounds and burns (bark tea, powder and/or ointment), and diverse products, such as shampoos, creams, capsules, soaps, etc., are commercialized. The bark is wrinkled, reddish-brown to grey, fibrous texture, 0.5-1.5 mm thick, resinous and with an astringent odor and flavor, and with a great quantity of tannins. The wood presents extremely short vessel elements, with alternate areolate punctuations, and simple perforated plates, vasicentric axial parenchima, confluent stripes, uniseriated rays, extremely short, fine and very short libriform fibres. The bark contains tannins, saponins, an alkaloide fraction, lipids, phytosterols, glucosides, xylose, rhamnose, arabinose, lupeol, methoxychalcones, and kukulkanins. PMID- 11487940 TI - Inclusion bodies induced by Bean rugose mosaic comovirus seen under light microscopy. AB - Two types of inclusion bodies were consistently observed under light microscopy in bean (Phaseolus vulgaris) leaf tissue infected with Bean rugose mosaic virus (BRMV), a species of the genus Comovirus, family Comoviridae. One type consisted of vacuolated inclusions found mainly in the cytoplasm of epidermal cells. The other type consisted of abundant crystalloid inclusions of different sizes and shapes found consistently in glandular hairs, guard cells, phloem tissue, xylem elements and occasionally in epidermal and mesophyll tissues. The two types of inclusion bodies stained with Azure A and Luxol Brilliant Green Bl-Calcomine Orange 2RS (O-G), and were similar to those seen to be caused by other species of comoviruses. PMID- 11487941 TI - [Space and time distribution along a succesional gradient of coprofagous coleopterans (Scarabaeinae, Aphodiinae) fauna in a tropical mountain forest, Narino, Colombia]. AB - Dung beetles (Scarabaeinae y Aphodiinae) were used to evaluate the effects of human activities in the area surrounding La Planada Natural Reserve, southwestern Colombia. During 1993 we monitored three habitats: primary and secondary forest, and open fields used for cattle grazing. A total of 55,296 trap/hours, evenly distributed among the three habitat types, captured 9,115 individuals (18 species, 11 genera). There were differences in species richness between habitats (F2,9 = 29.88; P < 0.001), an in number of individuals (F 2,9 = 36.22; P < 0.001). Collecting sites differed within habitats. Cluster analyses show that species composition differs between habitats with and without tree cover. Open areas act as barriers for movements of forest species. Differences between habitats and collecting sites may reflect high environmental heterogeneity, land use history of the sites or the influence of nearby disturbance. Some of the species found in open fields come from lower elevations and are usually associated with intense human disturbance. The proportion of digging and rolling species is similar in both primary and secondary forest, nocturnal species are more abundant; in open areas rolling species are absent and the number of diurnal species increases. We found no relationship between rainfall and species richness (r2 = 0.26; P = 0.41), nor between rainfall and number of individuals collected throughout the year (r2 = 0.07; P = 0.84). For all species the number of individuals collected decreased during dry season and during the onset of the rainy season. PMID- 11487942 TI - The species of Phora (Diptera: Phoridae) of Costa Rica. AB - The Phora of Costa Rica were reviewed and three species, all belonging to Schmitz's Group III, were recognized: P. americana Schmitz and Wirth, P. truncata new species and P. paramericana new species. Of the three species, P. americana is by far the most abundant, with the other two species being rarely collected. All specimens were collected in the central highlands of the country. The previous record of the Holarctic Region species P. stictica Meigen from Costa Rica was found to be based on a misidentification of P. truncata. PMID- 11487943 TI - Forager size of the leaf-cutting ant Atta sexdens (Hymenoptera: Formicidae) in a mature eucalyptus forest in Brazil. AB - We determined the size characteristics of foragers of the leaf-cutting ant Atta sexdens in a mature eucalyptus forest in Campos dos Goytacazes, Rio de Janeiro State, Brazil, at daytime (7:30 to 10:00 hr) and nighttime (19:00 to 23:00 hr). There were no significant differences between daytime and nighttime ant mass (Ma), but leaf fragment mass (Ml) and burden (B = [Ma + Ml]/Ma), which indicates relative load capacity, were significantly greater at daytime. There was a positive linear relationship between Ma and Ml for the combined daytime and nighttime data, and increases in Ma resulted in lower B. We compared A. sexdens characteristics with published results for Atta cephalotes, a closely related species. A. sexdens is larger and therefore able to carry heavier loads, but its burden is about 72% of the average value for A. cephalotes. We suggest that the lower load capacity of A. sexdens in comparison to A. cephalotes is related to its relatively larger size. PMID- 11487944 TI - Egg parasitoids of Podisus sculptus distant (Heteroptera: Pentatomidae) in an Eucalyptus plantation in the Brazilian amazonian region. PMID- 11487945 TI - A new species of rainfrog of the Eleutherodactylus milesi species group (Anura: Leptodactylidae) from Guatemala. AB - A new rainfrog of the Eleutherodactylus milesi group is described from the Sierra de Xucaneb, Guatemala. It is compared to the other 11 upland species from southern Mexico, Guatemala and Honduras referred to the group. Within the milesi group the new species differs from the other five forms which also have toe fringes in size of the finger disks, the snout shape in profile and the amount of toe webbing and dorsal tuberculation. Difficulties in distinguishing among the character states for the tympanum (distinct, indistinct and hidden) and between toe ridges and fringes indicate that very subtle differences separate recognized species. PMID- 11487946 TI - Care and cleaning of the endoscope. AB - The purchase of veterinary endoscopy equipment requires a large investment on the part of the hospital. To ensure good return on the investment as well as to provide adequate service to clients, the equipment should be properly maintained. Close attention to this detail enables the practitioner to extend the life of the endoscope and its accessory instruments. PMID- 11487947 TI - Esophagoscopy. AB - Esophageal pathology is one of the areas that had the major benefits from the advent of endoscopy. Esophagoscopy is a highly reliable diagnostic method for evaluating esophageal disorders that affect the mucosa or alter the lumen of the organ, such as foreign bodies, esophagitis, reflux disease, strictures, ulcers, fistula, and neoplasia. With endoscopy, the treatment of esophageal disorders has greatly improved as well, with the retrieval of foreign bodies and the dilation of esophageal strictures under direct visualization the main therapeutic indications. PMID- 11487948 TI - Gastroduodenoscopy in the dog and cat. AB - Endoscopic examination of the upper gastrointestinal (GI) tract in dogs and cats is a relatively recent development. It was just over 20 years ago, in 1978, that the first series report of GI endoscopy in dogs and cats was presented. In those initial days, endoscopy was truly a novelty, and the primary uses for endoscopes were to retrieve foreign objects or visualize the esophagus and stomach. It was not until the mid-1980s that technology and training made gastroduodenoscopy standard practice. The evolution in endoscopic technology has also mirrored the evolution in computers and imaging modalities. The original fiberoptic endoscopes, although still available, are giving way to video endoscopy and digital imaging capabilities that provide spectacular images and an ability to generate hard copy images for medical records, teaching purposes, and research data that was previously not possible. PMID- 11487949 TI - Colonoscopy, proctoscopy, and ileoscopy. AB - Colonoscopy and proctoscopy are primarily performed on dogs and cats with signs of chronic large bowel or rectal disease, and ileoscopy is typically performed in patients with signs of either large or small bowel disease. These techniques should not be used indiscriminately but only on animals that have a reasonable chance to benefit from their use. Even then, the techniques must be performed carefully so that the results are meaningful instead of misleading. We first consider what does and what does not constitute a reasonable indication for endoscopic examination of the lower intestines. After that, we turn to the specific techniques involved. PMID- 11487950 TI - Rhinoscopy, pharyngoscopy, and laryngoscopy. AB - Rhinoscopy, pharyngoscopy, and laryngoscopy are important procedures to include in the workup of patients with upper airway disorders. They are simple to perform, yield important information, spare the patient from undergoing invasive rhinotomy, and improve client relations when a diagnosis is reached. PMID- 11487951 TI - Small animal bronchoscopy. AB - Bronchoscopy is a useful diagnostic and therapeutic tool in veterinary medicine. The increased availability of fiberoptic technology and video-endoscopy has enhanced recognition of the benefits gained from visualization of the lower airways in animals with lower respiratory tract disease. Specimens retrieved from the lower airway during bronchoscopy have greater diagnostic capacity, and a better understanding of the pathophysiology of disease is provided through application of bronchoscopy to animals with respiratory tract disease. PMID- 11487952 TI - Laparoscopy in dogs and cats. AB - Laparoscopic techniques have been developed in veterinary medicine for noninvasive evaluation of several organs, including liver, extrahepatic biliary system, pancreas, kidneys, spleen, bowel, and genitourinary tract. Biopsy procedures of these organs with minimal risk can be performed in most patients. This article reviews the equipment and technique necessary to perform laparoscopy in dogs and cats. PMID- 11487953 TI - Video-assisted thoracoscopy. AB - Thoracoscopy is a minimally invasive operative endoscopic procedure designed for visual inspection of the thoracic cavity. The use of small, highly maneuverable, rigid telescopes allows visualization beyond what is possible by standard operative techniques. The use of a small video camera attached to a standard rigid telescope allows the operator and assistants to view a simultaneous, enlarged, and clear image. This article focuses on the basic diagnostic and surgical techniques used in veterinary video-assisted thoracoscopy. PMID- 11487954 TI - Arthroscopy. AB - Canine arthroscopy has made significant technological and scientific advancements in the last 10 years. The history of canine arthroscopy and currently available equipment required for performing diagnostic and therapeutic arthroscopy are discussed. Basic arthroscopic techniques are described in detail. Specific approaches to the major joints and associated disease processes treatable by arthroscopic methods are reviewed. The future of canine arthroscopy is bright and specific procedures for treating the joint diseases of dogs will undoubtedly be described as the collective arthroscopic expertise of veterinary surgeons increases. PMID- 11487955 TI - Uroendoscopy. Evaluation of the lower urinary tract. AB - Recent advances in uroendoscopy have allowed diagnostic evaluation of the lower urinary tract in most of our canine and feline patients. By providing a magnified view of the luminal surfaces of the lower urinary tract, uroendoscopy provides useful diagnostic information that is not readily available even by more invasive techniques. PMID- 11487956 TI - Uses and indications for video-otoscopy in small animal practice. AB - Acute and chronic otitis externa and otitis media are common disorders in dogs and cats. In combination with other diagnostic and therapeutic procedures, the video-otoscope is a useful and effective tool in the management of clinical cases. The enhanced illumination and magnification provide the practitioner with more detailed information for diagnosis and prognosis, and the configuration of the working channel facilitates sampling, improves efficacy of cleaning procedures, and decreases risks of iatrogenic injury to structures of the middle and inner ear. Photographic documentation of clinical cases enhances the medical record, communication with colleagues, and client education. Although video otoscopy facilitates diagnosis and therapy, it does not replace other important diagnostic tests such as evaluation for atopy, adverse food reactions, and immune compromising disease. Failure to identify the underlying primary cause usually results in treatment failure regardless of the technology employed. PMID- 11487957 TI - [Limits can also be strengths!]. PMID- 11487958 TI - The impact of casemix on the care of the elderly. AB - The Australian Health care system is a mix of public and private provision. The Federal Government funds medical care and the pharmaceutical benefit scheme while the State Governments are responsible for funding the public hospitals. Geriatric Medical care is provided in the public hospital system. The Australian DRG system has evolved to more adequately explain illness severity by a greater use of the complications and comorbidities. The structure of the Sub-Acute and Non-Acute Patient (SNAP) classification is outlined. While it is anecdotally said that the introduction of DRG-based funding is detrimental to the elderly, the published evidence does not support this. The potential benefits of a casemix system are discussed. PMID- 11487959 TI - [AR-DRG effects on geriatrics in Germany]. AB - The parties of the self-governing Health Service are at present preparing the introduction of a general case, flat rate compensation system for the hospital section on behalf of the legislature. The system is based on the performance definitions and case classifications of the Australian DRG System, Version 4.1. The relative evaluation of the individual DRG for the implementation into the system in Germany will be based on German cost and benefit data. For this purpose a calculation of case-specific treatment costs based on a representative selection of hospitals will be made. The geriatrics field will have to adapt itself to these regulations in the same way as all other medical disciplines. Difficulties are arising due to the inconsistent classification of cost and benefit data in connection with the classification. The geriatric field is under represented in the calculation of relative evaluation and, therefore, the spectrum of benefit cannot be shown in the planned calculation system. Analogous to the calculation of German relative cost evaluation, the geriatric section will define in a project the actual costs for their treatment cases and determine the consecutive relative evaluation. PMID- 11487960 TI - [Clinical laboratory diagnosis and aging. 3: Evaluation of a study of aging- complete blood and urine status]. AB - The results of the determination of hematologic values from 262 men and 242 women, participants of an aging study and half of each group 44.4 +/- 0.9 and 63.0 +/- 0.9 (men) and 44.4 +/- 0.9 and 62.8 +/- 0.8 years old (women), respectively, are compared. In men, one analyte (hemoglobin decreasing) and four indices show significant differences (MCV increasing, MCH decreasing, MCHC decreasing, RDW increasing). In the older group, the iron level and the transferrin saturation are also significantly lower. In women, erythrocytes and the hematocrit are significantly higher in the older group whereas the indices MCH and MCHC are lower and the RDW increases. At the same time, the iron level, transferrin and the transferrin saturation decrease whereas ferritin doubles. The sex differences of the hematologic parameters are more pronounced in the younger participants and especially remarkable in ferritin in both age groups. The results of the semiquantitative analysis of ten urine parameters by reagent strip show differences with respect to sex (e.g., leucocytes and erythrocytes) and age (e.g., specific gravity, pH, nitrite, protein, erythrocytes). The usefulness of the estimation of glucose in urine is discussed in connection with the corresponding serum glucose levels. PMID- 11487961 TI - Secondary prevention of breast cancer in older women. AB - Periodic mammography is well studied and widely applied as a screening programme to reduce breast cancer-related mortality and morbidity in women aged 50 to 69 years. Despite the fact that age is a major risk factor for breast cancer, no evidence-based data are available on survival benefit of screening in women older than 69 years. The most commonly cited guidelines for screening in breast cancer disagree on the upper age limit of the target population. This age limit is a matter of cost-effectiveness and is influenced by active life expectancy, risk for breast cancer, comorbidity and functional status. Benefit of screening also depends on adherence rate of elderly women in screening programmes and optimal treatment of identified tumours. In a selected population of elderly women, screening for breast cancer might be cost-effective. PMID- 11487962 TI - [Test-retest reliability of a German language multidimensional assessment instrument in elderly probands]. AB - Most geriatric assessment instruments have been developed in the English language. Translated versions might differ in their psychometric properties. We analyzed the test-retest reliability and internal consistency of a German instrument for multidimensional geriatric assessment that was based on a newly developed English version. A group of 100 over 75-year-old community-dwelling persons (mean age 83.0 years, 81% women) in Hamburg (n = 26) and Ulm (n = 51), Germany, and Berne (n = 23), Switzerland was interviewed twice by the same trained interviewers with a one week interval. We administered questions on general health, chronic disorders, basic and instrumental activities of daily living, urinary incontinence, nutrition, falls, pain, the social support/network and preventive care measures. In addition, the Functional Status Questionnaire, the Physical Activity Scale for the Elderly, the Geriatric Oral Health Assessment Index, the Visual Function Questionnaire, the Hearing Handicap Inventory for the Elderly and the Geriatric Depression Scale were administered. Cohen's kappa was good to excellent (0.64 < or = kappa < or = 0.89) with only three exceptions (pain questions, kappa = 0.53; questions on preventive care services, kappa = 0.51; and one of the questions on recent falls, kappa = 0.44). Cronbach alpha (internal consistency) was good to excellent for all domains (0.76 < or = alpha < or = 0.95). The study results confirm good test-retest reliability of the German version of this multidimensional geriatric assessment instrument. Adapted versions of this instrument can be used for different purposes, e.g., preventive home visits, outpatient geriatric assessments or epidemiological studies in older persons. PMID- 11487963 TI - Families and intergenerational relationships in aging societies: comparing the United States with German-speaking countries. AB - Industrialized nations have experienced many demographic changes favoring older adults during the last few decades of the 20th century. This article is a comparative examination of current and future trends in intergenerational relationships and aging in two societies: the United States and German-speaking countries. While comparing similarities and significant differences between the cultures, we examine five issues related to these trends for families and intergenerational relationships in the 21st century: (1) consequences of population aging for family structures and relationships--the emergence of the beanpole family structure and the longer years of linked lives across generations; (2) the debate about the decline of the family; (3) intergenerational solidarity over the life course; (4) intergenerational conflict across the life course; (5) prospects for solidarity and conflict between generations in the 21st century. Similarities between cultures include the demographic realities of population aging, the shift from pyramids to beanpoles, more years of shared lives between and across generations, more non-traditional family forms, and intergenerational conflicts in families. Differences include diverse replacement fertility rates, alternative approaches to governmental supports for families and aging members, and possible differences in cultural values regarding individualism. PMID- 11487964 TI - [Significance of employment and unemployment in middle and advanced adult age for subjective perception of aging and realization of potentials and barriers of a responsible life]. AB - This contribution proceeds on the assumption that occupational activities in middle and upper adulthood do offer opportunities to use and extend competences and therefore to experience self-efficacy, social participation and acceptance, whereas unemployment is associated with a constriction of personal resources which have a negative impact on subjective perceptions of ones's own aging process, potentials and barriers of leading a productive and responsible life. First, employed and unemployed subjects are compared in their scores on three psychometric scales (perceptions of age-related losses, potentials and barriers of leading a productive and responsible life) and four aspects of the "Lebenslage" (Subjective health status, Perceived social integration, Income of the household, Education). In a further step the predictive power of employment and unemployment for subjective perceptions of one's own aging process, potentials and barriers of leading a productive and responsible life when aspects of the "Lebenslage" and sociostructural variables (Age, Sex, Eastern versus western part of Germany, Areas with high versus low unemployment) are also taken into account. Results, obtained in a stratified sample of 641 subjects between 45 and 57 years, show that employed and unemployed subjects differ in subjective perceptions of one's own aging process, potentials and barriers of leading a productive and responsible life as well as in aspects of the "Lebenslage". Employment and unemployment are significant predictors of subjective perceptions of one's own aging process, potentials and barriers of leading a productive and responsible life even when aspects of the "Lebenslage" and sociostructural variables are already taken into account. Results are interpreted as support for the hypothesis that occupational activities are not only instrumental for the realization of human necessities, but that in middle and upper adulthood employment is to be regarded as a human need in itself. PMID- 11487965 TI - [Subjective cognitive dysfunctions in the elderly: a change-oriented viewpoint]. AB - The relation between subjective cognitive complaints and the actual level of cognitive performance in the elderly is of interest in both basic and applied cognitive research. However, most studies suggest a small to moderate correlation between complaints and actual performance. In contrast to previous studies, we examined the relation between changes in fluid and crystallized intelligence, changes in memory performance, and changes in subjective cognitive complaints in a sample of 465 older adults (M = 62.9 years) at two measurement occasions four years apart. The results indicate significant relationships between changes in subjective complaints and changes in memory performance and changes in fluid intelligence. The discussion focuses on the importance of a change-oriented view in examining subjective cognitive complaints from an applied perspective. PMID- 11487966 TI - [In memory of Gustav A. Lienert]. PMID- 11487967 TI - Synthesis and bovine beta 3-adrenergic agonistic activities of a novel series of aryloxypropanolamines. AB - We synthesized a novel series of 21 aryloxypropanolamine compounds characterized by N-alkyl, aralkyl, and aryl substituents. The compounds showed potent beta 3 adrenergic agonistic activities in Chinese hamster ovary cells expressing the bovine beta 3-adrenoceptors with Kact and Ki values of 4.2 +/- 3.0 nM and 459 +/- 169 nM respectively, for the ligand with the best compromise between potency and affinity. Structure-activity relationships are discussed. PMID- 11487968 TI - Synthesis and modulation of cytokine production by two new adamantane substituted acyclic desmuramyldipeptide analogs. AB - Two new adamantyl-desmuramyldipeptides LK 415 and LK 517 with 1 adamantylcarboxamido moiety as a replacement for muramyldipeptide's N acetylglucosamine fragment were synthesized. Their efficacy to modulate the production of cytokines was measured in vitro in ionomycin and phorbol-12 myristate-13-acetate (PMA) activated cultures of human peripheral blood mononuclear cells (PBMC), co-incubated with the substances tested. The results were compared with the activity of muramyldipeptide (MDP). All three substances are strong up-regulators of IL-12 synthesis and hence of the IFN gamma synthesis as well. While MDP and LK 415 are relatively ineffective in modulation of IL-2, IL-4 and IL-10 production in vitro, the synthesis of all three cytokines is considerably up-regulated when peripheral blood mononuclear cells are co incubated with LK 517. It seems likely that the introduction of the diethyl phosphonate moiety into LK 517 is of great importance for the augmented T-cell cytokine production. PMID- 11487969 TI - [Partially hydrogenated aryl-1,2/1,4-anthraquinone derivatives, 5-lipoxygenase inhibitors with arotinoid structure]. AB - The combination of 5-LOX inhibition and retinoid activity in one molecule could be an interesting pharmacological tool to influence psoriasis. Thus we synthesized compounds with arotinoid structure by anellation of the 5-LOX inhibitors 1 and 2 with 1,1,4,4-tetramethylcyclohexane. A key step was the CuCl MeCN-O2 oxidation of tetrahydroanthracenol 13 to the corresponding 1,2 anthraquinone 14 which could be converted to the analogous 2-hydroxy-1,4 anthraquinone 19 by Thiele-Winter reaction followed by oxidation. The halogenated quinones 9 and 21 were arylated with 2,6-di-tertbutylphenol and demethylated or hydrolyzed to the target compounds 3 and 4 which were tested in comparison with the non-anellated 5-LOX inhibitors 1 and 2 for LOX inhibition in activated human granulocytes and for antioxidative activity by the method of Popov with the chemiluminometer Photochem. The results are discussed in relation to the corresponding logP values. The 1,2-quinones 1 and 3 are more potent 5-LOX inhibitors than their 1,4-analogues 2 and 4, the tetrahydroanthraquinon derivatives 3 and 4 are less potent than the naphthoquinones 1 and 2. All compounds are devoid of any activity in cell differentiation as compared to retinoic acid as indicated by the NBT test with HL-60 leukemia cells. PMID- 11487970 TI - Synthesis of 1,2-unsaturated pyranosylphosphonate nucleosides from 3,4,6-tri-O acetyl-D-glycal. AB - Ferrier rearrangement of 3,4,6-tri-O-acetyl-D-glucal (1) in the presence of triethylphosphite afforded the 2,3-unsaturated pyranose 2. Deacetylation and simultaneous migration of the double bond to 1,2-position in the sugar moiety was achieved by stirring in sodium ethoxide. Tosylation with one equivalent of tosyl chloride afforded 4. Nucleophile displacement of the tosylate of 4 with nucleobase in the presence of NaH/DMF followed by deprotection gave the desired 1,2-unsaturated pyranosylphosphonates 7a-c. PMID- 11487971 TI - Chiral separation of verapamil and some of its metabolites by HPLC and CE. AB - HPLC and CE assays were developed for chiral separations of verapamil and its metabolites in serum samples. Three chiral HPLC columns (Chiralcel OJ, Chiralpak AD and Chiralcel OD-R) were tested in normal and reverse-phase modes. All HPLC analyses were performed with fluorescence detection at 276 and 310 nm. CE was realized using CM-beta-CD as a chiral selector for the enantiomeric analysis. The results of HPLC and CE studies were compared and the possibilities for the applications in therapeutic drug monitoring were discussed. PMID- 11487972 TI - Determination of molecular sulphur in sodium thiosulphate injection solutions by high performance liquid chromatography. AB - A method was developed to quantify the sulphur formed in decomposed sodium thiosulphate injection solutions. Elemental sulphur was extracted with cyclohexane (LLE) and analysed by RP-HPLC. A linear calibration curve with a detection limit of 24 micrograms/ml was obtained. Different amounts of sulphur were detected in extemporaneous and commercial solutions. PMID- 11487973 TI - Identification of lactate. Analytical methods of pharmacopoeias with DBH in respect to environmental and economical concern, Part 4. AB - The identification of lactate according to Ph. Eur. 1997 and DAB 2000 uses the oxidation of lactic acid to pyruvic acid by boiling with bromine water in sulphuric acid. Acetaldehyde arising by decarboxylation is detected according to Legal applying a time consuming and troublesome procedure. 1,3-Dibromo-5,5 dimethylhydantoin (DBH) as well as potassium bromate can replace elemental bromine. Lactic acid and all lactates of Ph. Eur. 1997 and DAB 2000 can be identified better and faster using lactate oxidase (LOD, test strip system Accusport). According to DAB 2000 the base of ethacridine lactate has to be separated. This is no longer necessary, if an enzymatic identification is applied. PMID- 11487974 TI - An improved method for the determination of betaine in Echinacea products. AB - A rapid and sensitive HPLC method for the separation and quantification of betaine in Echinacea products has been developed. Strong cation-exchange (SCX) material was used as stationary phase, and a mixture of methanol and 50 mM choline buffer (pH 3.5) as mobile phase. After formation of the bromophenacyl derivative, betaine was detected at 254 nm with a detection limit of 0.2 microgram/ml. The method was successfully used to analyze several Echinacea market products, and significant variations in their betaine content from 0.04 to 0.64% were observed. PMID- 11487975 TI - Preparation, characterization and in vitro antimicrobial activity of metronidazole bearing lectinized liposomes for intra-periodontal pocket delivery. AB - Liposomes constructed of egg phosphatidylcholine (EPC), cholesterol (Chol) and stearoylamine (SA) were coated with lectin (Concanavalin-A). These lectinized liposomes were found to retain the ligand binding activity of surface coated concanavalin A (Con-A) as demonstrated by bovine submaxillary mucin (BSM) binding assay. Moreover the ligand specificity of Con-A was maintained even after coating the liposome surface because the presence of competing sugar alpha-methyl mannoside, significantly inhibited the interaction of lectinized liposomes and BSM. The significance of divalent cations for these interactions was studied. The Con-A coating was found to be stable in simulated salivary fluids (SSF, pH 7.2) and under various pH conditions. In vitro targeting studies of lectinized liposomes with gram-negative bacilli (Streptococcus mutans) that harbor in the periodontal pocket (biofilm) demonstrated nearly 100% bacterial growth inhibition (% BGI). The antimicrobial effect was maintained for 360 min. The results were compared with metronidazole bearing plain (protein free/uncoated) liposomes and the free drug at the same dose levels. Mechanisms involved are also discussed. These observations suggest that liposomes coated with lectin (Con-A) were able to maintain the sugar affinity and specificity of the associated ligand and could be targeted to the surface 'glyco-calyx' of bacterial bio-film. PMID- 11487976 TI - Experimental data and theoretical considerations concerning the validity of the SAL concept to characterize non-thermal antimicrobial treatments. AB - For sterilization processes the pharmacopoeias demand a sterility assurance level (SAL) of 10(-6), i.e. a probability of not more than one viable microorganism among one million sterilized products. This SAL concept is based on the assumption that the inactivation of microorganisms by physical or chemical means generally follows first-order kinetics. In this paper it is demonstrated that this is not absolutely true for non-thermal antimicrobial processes. Using Bacillus subtilis spore test preparations the sporicidal efficacy of gamma and ultraviolet irradiation on the one hand as well as the treatment by glutaraldehyde and hydrogen peroxide containing solutions on the other hand was investigated. A range of mean spore contamination between 10(8) and 10(-2) spores per test item could be supported by experimental data. It was demonstrated that the antimicrobial treatment parameters which are sufficient to reduce a high spore burden were not valid for an adequate reduction of the remaining lower spore burden. It is concluded that any extrapolation of such experimental data to the SAL range as usual in the validation of sterilization process parameters may be not permitted. Possible theoretical explanations of the non-homogeneity of the spore inactivation by non-thermal methods as well as consequences for the safety evaluation of sterilization processes are discussed. PMID- 11487977 TI - Pharmacological properties of some aminoalkanolic derivatives of xanthone. AB - A series of appropriate aminoisopropanoloxy derivatives of 2-, 3- or 6-xanthone was synthesized and evaluated for anticonvulsant activity in the maximal electroshock seizure (MES) and subcutaneous pentylenetetrazole seizure threshold (ScMet) assays and for neurotoxicity (TOX). The most interesting result was the anticonvulsant activity of (+/-)-3-(2-propylamino)-1- [(2-methyl)-6-xanthonoxy]-2 propanol hydrochloride (10), which displayed anti-MES activity with a protective index (TD50/ED50) of 0.80. Some of the obtained compounds were also tested for their effect on the circulatory system (influence on the non-working heart perfusion, protection against adrenaline induced-arrhythmia) and acute toxicity. PMID- 11487978 TI - Profisetinidin type tannins responsible for antioxidant activity in Copaifera reticulata. AB - The in vitro antioxidant and free radical scavenging properties in bark extracts of South American tree Copaifera reticulata Ducke. (Caesalpinaceae) were studied using different bioassays. Lipid peroxidation was assessed by means of the production of thiobarbituric acid reactive substances (TBARS) in rat liver homogenate. All the extracts tested were effective in this method. The highest activity was observed in the aqueous extract, showing an IC50 of 30 micrograms/ml. DNA sugar damage induced by Fe (II) salts was also used to determine the capacity of the samples to suppress hydroxyl radical-mediated degradation of DNA. Although all the extracts tested were effective in reducing oxidation of DNA, the highest activity was observed in the methanol extract, showing an IC50 of 2 micrograms/ml. Bioassay-guided fractionation of a total methanol extract monitored by luminol-enhanced chemiluminescence, together with structural elucidation using 13C NMR and FABMS, led to the identification of profisetinidin type tannins in a semi-pure fraction. The fraction containing the active compounds also reduced the production of TBARS in rat liver homogenates (IC50 = 530 micrograms/ml) and DNA damage (IC50 = 1 microgram/ml), suggesting that profisetinidins could be responsible for the free radical scavenging and antioxidant activities observed in the extracts. PMID- 11487979 TI - Isolation and structure elucidation of viscoazucine, a novel sesquiterpene from Polygonum viscosum. AB - A new sesquiterpene, 1,4-dimethoxycarbonyl-7-(1-methylethyl)- 3,3a,6,7,8,8a hexahydroazulene (viscoazucine) (1), and a known flavone, 3',5,7-trihydoxy 3,4',5'-trimethoxyflavone (4), have been isolated from Polygonum viscosum. The structures of these compounds were elucidated by spectroscopic analyses, notably UV, MS and NMR. PMID- 11487980 TI - Two new flavonol glycosides from leaves of Koelreuteria paniculata. AB - Two new flavonol glycosides were isolated from dried leaves of Koelreuteria paniculata Laxm. (Sapindaceae) and characterized as 6,8-dihydroxy-afzelin and afzelin 3"-O-gallate, based on chemical and spectral evidences, in addition to nine known polyphenolic metabolites, including eight isolated for the first time from this species. PMID- 11487981 TI - The history of electrically-assisted transdermal drug delivery ("iontophoresis"). AB - The first proposals for the use of electric current in drug delivery date from the mid 18th century. Serious progress was made in the 19th century notably by Benjamin Ward Richardson (1828-1896), Hermann Munk (1839-1912), William James Morton (1846-1920), Stephane Leduc (1853-1939) and Fritz Frankenhauser (born 1868). Administration of metal ions as well as alkaloids was tried at that time. Until the early 20th century, current mediated drug delivery was known as "cataphoresis"; Frankenhauser is said to have introduced the term "iontophoresis" before 1908. Recently, researchers talk about "electrically-assisted transdermal drug delivery". The technique was never widely adopted but always proved useful to some extent in solving particular drug delivery problems. At the dawn of the 21st century, attempts are being made to achieve iontophoretic delivery of peptides and proteins. PMID- 11487982 TI - Ampicillin prodrugs: amide conjugates from amino acids and ampicillin. PMID- 11487983 TI - A validated gas chromatographic method for the determination of isosorbide-5 mononitrate in human plasma and its application to pharmacokinetic studies. PMID- 11487984 TI - Mixing of pharmaceutical powders in tablet manufacture. PMID- 11487985 TI - Emulsions as oral moisturisers for the treatment of severe xerostomia. PMID- 11487986 TI - The Health Insurance Portability and Accountability Act (HIPAA) and physician compliance. AB - The Administration and Simplification Provisions and the Standards for Privacy of Individually Identifiable Health Information Section of HIPAA (Health Insurance Portability and Accountability Act) will have a significant impact on the daily operations of most physicians' practices. The HIPAA legislation is summarized, and these two sections of HIPAA are discussed in detail. The rationale for the regulations, their substance and intent, and physician compliance strategies are discussed. PMID- 11487987 TI - American Medical Association lawsuits against managed care organizations. PMID- 11487988 TI - Role reversal? PMID- 11487989 TI - Advancing years. PMID- 11487990 TI - Effects of a stage-based alcohol preventive intervention for inner-city youth. AB - The purpose of this study was to examine first year outcomes of an alcohol preventive intervention within inner-city middle schools. Subjects consisted of 650 sixth grade students from one neighborhood inner-city school (n = 262) and one bused school (n = 388). At posttest, chi-square analyses showed that significantly fewer neighborhood intervention students initiated alcohol use, used alcohol during the past seven-day and thirty-day periods, drank heavily during the past thirty days, and drank over any period of time, compared to control students (p's < .05). Significant group x prior alcohol consequences interaction effects were found for bused students, showing those with past alcohol consequences who received the intervention had less intentions to use alcohol and less frequent use of alcohol (p's < .05). These results suggest that a brief, stage-based preventive intervention may result in significant reductions in alcohol initiation and consumption among some inner-city youth. PMID- 11487991 TI - Constructing a consensus-based prevention outcome measurement instrument. AB - This article describes Ohio's Prevention Evaluation Project (PEP). The purpose of the project was to develop a process and instrument to assess behavioral and attitudinal outcomes in primary drug abuse prevention programs targeting young people aged twelve to seventeen. One of PEP's principal achievements was the inclusion of community prevention program providers from throughout the state in the evaluation instrument development. The effort produced a self-administered questionnaire to capture data on young peoples' drug use practices, attitudes toward drugs, and problematic behaviors. Most significantly, the product produced was the acceptance and endorsement of community-based programs. The forty-one item questionnaire has good to excellent reliability for virtually all measures, as assessed by the test-retest method. The instrument is a product of a process that brought together a university-based research team, state alcohol and drug abuse prevention administrators, and representatives of public programs. PEP demonstrates how to productively link service providers, administrators, and evaluators to develop a process for assessing the effectiveness of prevention programs. The process and tools described serve as a model for other communities wanting to evaluate their prevention programming. PMID- 11487992 TI - Do training programs work? An assessment of pharmacists activities in the field of chemical dependency. AB - OBJECTIVES: The primary objective of this study was to determine if a selected sample of pharmacists who had attended a chemical dependency training program were performing more chemical dependency related activities than a group of American Pharmaceutical Association (APhA) members who had not had this training. Additionally, an assessment of the perceived barriers to performing chemical dependencyrelated activities was performed. DESIGN: A confidential mail questionnaire was sent to 305 Utah School participants and 305 APhA members who had not participated in the program (See Appendix). RESULTS: Respondents who had received educational training in chemical dependency were more likely to perform the following activities: lecture to community groups and health care professionals about chemical dependency, participate in a pharmacists' recovery program, provide patients with information about treatment centers, and counsel patients about the alcohol in over-the-counter products. Respondents who had not received chemical dependency training indicated that the following barriers prevented them from taking a more active role in the chemical dependency field: lack of knowledge of chemical dependency resources in the community, unaware of how to get involved with the state recovery program, belief that involvement in the state recovery program would hurt their professional reputation, belief that chemically dependent individuals cannot be rehabilitated, and uncomfortable working with chemically dependent patients. CONCLUSIONS: Pharmacists who have attended substance abuse training programs are performing more chemical dependency activities than pharmacists who have not received training in chemical dependency. In addition, different barriers to performing chemical dependency related activities exist between pharmacists with and without training in this field. PMID- 11487993 TI - Outpatient cocaine abuse treatment: predictors of success. AB - In 1990 more than 238,000 patients in the United States entered treatment, with cocaine as their primary diagnosis. This represented 36 percent of the treatment admissions for substance abuse. This research project uses data from the U.S. Army's Alcohol and Drug Abuse Prevention and Control Program to analyze which treatment and demographic variables have an effect on cocaine treatment outcomes. The sample included 3602 soldiers entered into treatment between 1983 and 1992 for cocaine abuse. The logistical regression results suggested the following treatment variables had an effect on outcomes: the type of treatment provided, the length of time in treatment, the number of different types of drugs being abused prior to treatment, and the length of time since the patient had abused cocaine prior to entering treatment. Race of the patient had an impact on treatment outcomes, but gender, educational level, age, rank of the soldier, marital status, and length of time in the military did not have an impact on treatment outcomes. PMID- 11487994 TI - Feedback from participants and leaders in an intervention program: the 1997-1998 opening doors process evaluation. AB - Parents, students, and program leaders involved in an in-school drug prevention program called Opening Doors were assessed for their perceptions of the efficacy of the program. In general, the feedback indicated a very high program satisfaction level from respondents with parents indicating the highest level satisfaction (92%). Areas for improvement ranged from: parent attendance, invitation process, increased support from schools, and increasing awareness of scheduling and time involvement by leaders and school administration based on program leader feedback as well as invitation process and program duration from the parents (e.g., 37% thought the program was too short). Student satisfaction may be predicted from the perceived impact of the program on significant relationships (e.g., with peers and family) as well as affective reactions to the program (e.g., enjoyment of the program). Finally, a school-level analysis indicated that the application of the Opening Doors Program in different schools was provided uniformly. PMID- 11487995 TI - Challenging the collegiate rite of passage: a campus-wide social marketing media campaign to reduce binge drinking. AB - A social marketing media campaign, based on a normative social influence model and focused on normative messages regarding binge drinking, on a large, southwestern university campus has yielded positive preliminary results of an overall 29.2 percent decrease in binge drinking rates over a three-year period. The Core Alcohol and Drug Survey and the Health Enhancement Survey provided information on student knowledge, perceptions, and behaviors regarding alcohol and binge drinking. This study represents the first in-depth research on the impact of a media approach, based on a normative social influence model, to reduce binge drinking on a large university campus and has yielded promising initial results. PMID- 11487996 TI - Efficacy of navigation in skull base surgery using composite computer graphics of magnetic resonance and computed tomography images. AB - The efficacy of a neurosurgical navigation system using three-dimensional composite computer graphics (CGs) of magnetic resonance (MR) and computed tomography (CT) images was evaluated in skull base surgery. Three-point transformation was used for integration of MR and CT images. MR and CT image data were obtained with three skin markers placed on the patient's scalp. Volume rendering manipulations of the data produced three-dimensional CGs of the scalp, brain, and lesions from the MR images, and the scalp and skull from the CT. Composite CGs of the scalp, skull, brain, and lesion were created by registering the three markers on the three-dimensional rendered scalp images obtained from MR imaging and CT in the system. This system was used for 14 patients with skull base lesions. Three-point transformation using three-dimensional CGs was easily performed for multimodal registration. Simulation of surgical procedures on composite CGs aided in comprehension of the skull base anatomy and selection of the optimal approaches. Intraoperative navigation aided in determination of actual spatial position in the skull base and the optimal trajectory to the tumor during surgical procedures. PMID- 11487997 TI - Lumbar disc herniation in patients up to 25 years of age. AB - Retrospective analysis of 165 patients (105 males, 60 females) with a mean age of 21.2 years (range 14 to 25 years) of 6933 surgically treated patients from January 1987 to May 1999 focused on age and sex distribution, body mass, familial predisposition, trauma, histology, and clinical course. The incidence of herniated lumbar discs was 2.3% in patients aged up to 25 years. A valid family history was obtained in 121 patients and a positive history was found in 82 of these patients (67.8%). The patients had a higher body mass index compared to a group of individuals with a similar age structure. Radiography demonstrated bony changes in 124 patients (75.2%), primarily attributable to postural deformities such as scoliosis. The condition of the bony structures seems to be more important than the condition of the disc tissue in the occurrence of this disease in young patients. PMID- 11487998 TI - Transorbital penetrating injury by a chopstick--case report. AB - A one-year-old boy presented with orbitocranial penetrating injury by a chopstick. Neurological examination did not reveal abnormal findings. Skull radiography did not reveal any sign of fracture and there were no abnormal findings. Initially, computed tomography (CT) of the head did not reveal any intracranial lesions. However, bone window CT showed a well-defined low-density abnormality measuring 2.5 cm in length in the right orbit and parasellar region. Magnetic resonance imaging clearly revealed a low-intensity structure extending from the orbit to the prepontine area. Surgical exploration was emergently performed and the wooden fragment was removed. The postoperative course was uneventful. Transorbital penetrating injury by a wooden foreign body is relatively rare. The wound may be superficial and trivial. Major neurological deficit does not usually manifest immediately, so the penetrating injury may be overlooked. If the foreign body is retained in the orbit and cranium, severe infectious complications may occur later. PMID- 11487999 TI - Cavernous angioma of the corpus callosum mimicking an astrocytic tumor--case report. AB - A 49-year-old female presented with a mass at the genu of corpus callosum manifesting as vertiginous sensation persisting for a couple of months. The preoperative diagnosis based on neuroimaging was astrocytic tumor, probably an oligodendroglioma. The mass was totally excised through a left interhemispheric approach without postoperative neurological deficits. Histological examination of the mass revealed only vascular components with tiny hemorrhage, compatible with cavernous angioma. Cavernous angioma at the genu of corpus callosum is very rare, and definitive diagnosis can be achieved only through surgery. Surgical intervention should be considered before starting radiotherapy without histological confirmation. PMID- 11488000 TI - Hearing restoration from deafness after resection of a large cerebellopontine angle meningioma--case report. AB - A 61-year-old man presented with a large cerebellopontine angle meningioma manifesting as a 1-year history of deafness on the right side, in whom hearing was restored from the deaf state immediately after tumor resection. Neuroimaging demonstrated a large mass in the right cerebellopontine angle, originating at the region adjacent to the jugular foramen. Audiometry showed his hearing was off scale (> 105 dB) on the right. The tumor was successfully removed through the retrosigmoid approach, and the integrity of the 7th and 8th cranial nerves was maintained. The patient regained hearing on the day after the operation, which continued to improve until near-normal. The 8th cranial nerve function may recover dramatically after removal of non-acoustic tumors, even if preoperative hearing loss is profound. To maximize the opportunity to regain hearing, approaches which devastate cochlear function should be avoided and more meticulous manipulation during tumor removal is needed. PMID- 11488001 TI - Mature teratoma arising from the sella--case report. AB - A 26-year-old, short statured, obese male presented with a mature teratoma located entirely within the dural confines of the sella manifesting as headaches and progressive loss of vision. He had panhypopituitarism. Magnetic resonance imaging showed a large sellar-suprasellar but entirely infradiaphragmatic tumor of varying consistency. The tumor was resected through a trans-sphenoidal route. The tumor had elevated the diaphragma sellae to a significant extent but did not pass through. Histological examination confirmed a mature teratoma. PMID- 11488002 TI - Solitary metastasis from occult follicular carcinoma of the thyroid mimicking trigeminal neurinoma--case report. AB - A 50-year-old woman presented with an extremely uncommon case of solitary metastasis from follicular carcinoma of the thyroid, which presented clinically as trigeminal neurinoma. Neuroimaging detected a tumor in the right petrous apex, which was removed surgically. Histological examination showed metastatic follicular carcinoma of the thyroid. However, no primary tumor was detected by various investigations. The tumor recurred twice, and was treated surgically both times. The patient finally agreed to adjuvant therapy for the suspected primary. Radiotherapy was performed followed by complete thyroidectomy. Examination of the gross specimen found the tumor nodule. Clinically significant metastasis can arise from histologically benign and silent follicular thyroid neoplasms. PMID- 11488003 TI - Reversible posterior leukoencephalopathy syndrome in a patient with hypertensive encephalopathy--case report. AB - A 58-year-old male presented with reversible posterior leukoencephalopathy syndrome (RPLS) manifesting as headache, papilledema, and renal hypertension. T2 weighted magnetic resonance (MR) imaging showed hyperintensity lesions in the medulla, pons, bilateral thalami, and bilateral deep white matter of the parieto occipital lobes. The pons was swollen. Diffusion-weighted MR imaging did not show increased intensity in these lesions. The lesions disappeared with improvement of clinical symptoms after treatment for hypertension. These findings suggested the lesions were vasogenic edema and the diagnosis was RPLS. T2-weighted and diffusion-weighted MR imaging are useful modalities to differentiate RPLS from other central nervous system abnormalities such as infarction, multiple sclerosis, and central pontine myelinolysis. The clinical and neuroradiological findings of RPLS can be reversed by timely initiation of treatment for the causative factor. PMID- 11488004 TI - [Dissociated horizontal deviations (DHD): nomenclature and etiology. An orientating attempt in terminological confusion]. AB - PURPOSE: To classify dissociated horizontal deviations (DHD) based on the etiology of the vergence that causes the dissociation of the squint angle. METHODS: Dissociated strabismus can reliably be diagnosed by a change of the squint angle caused by an alternation in the fixation from one eye to the other, when all the other conditions (head posture, direction of gaze, fixation distance, accommodation) remain unchanged. The decisive diagnostic tool is the reversed fixation test: During monocular fixation with one eye, the squint angle of the other eye is neutralized using a synoptometer or a prism. Then, the fixation is changed to the other eye, which keeps its position behind the prism. If the change in fixation causes a change in the position of the previously fixating eye, the deviation is dissociated. If the position of the previously fixating eye does not change, the deviation is not dissociated. RESULTS: Dissociated strabismus can be detected and differentiated from both comitant and incomitant non dissociated strabismus by the reversed fixation test. The dark red glass test is less sensitive and less specific for dissociated strabismus than the reversed fixation test, if a difference in accommodative convergence is excluded, possible causes of DHD are nystagmus-attenuating convergence, substitution of version eye movement by convergence, and a horizontal side effect of the vertical eye muscles in asymmetric DVD. CONCLUSION: Dissociated strabismus is an effect of vergence which is brought about by the change in the fixation or dominance from one eye to the other eye or in the interocular ratio of afferent input (luminance). Dissociated strabismus can be classified based on its direction or due to its etiology. PMID- 11488005 TI - [Near vision after implantation of monofocal versus multifocal intraocular lenses]. AB - AIM: Object of this study was to compare the near visual acuity between eyes with monofocal intraocular lens (MONO) and myopic target refraction in comparison with eyes after refractive multifocal intraocular lens (MIOL) implantation and emmetropic to slight hyperopic target refraction. PATIENTS AND METHOD: 89 eyes of 83 patients underwent cataract surgery, in 49 eyes a MONO (SI-40NB, Allergan) was implanted, in 40 eyes a refractive MIOL (SA-40N, Allergan). At day 2 after surgery and after 5 months the uncorrected near visual acuity, the near visual aculty with distance correction and with an addition of +3 diopters (D) were determined. The refractive outcome targeted in case of MIOL implantation was emmetropia to slight hyperopia and myopia for the MONO subjects. The eyes with MONO were classified in 4 categories depending on their postoperative refractive error (spherical equivalent, SE): category 1 with SE +1 to > 0 D, category 2 with SE 0 to > -1 D, category 3 with SE -1 to > -2 D and category 4 with a SE -2 to > 3 D. RESULTS: We found a statistically significant better uncorrected near visual acuity in eyes with MIOL than in MONO subjects for category 1-3. In category 4 there was no statistically significant difference between MONO and MIOL. With best distance correction near visual acuity in MIOL was statistically significant better than in all 4 MONO categories. After adding 3 D, there was no statistically significant difference between the groups. CONCLUSION: Even in case of a myopic target refraction near visual acuity in eyes of the MONO group was statistically significant worse than in the eyes of the MIOL group. Only in case of postoperative SE of < or = -2 D eyes of the MONO group had a near visual acuity similar to the eyes of the MIOL group. This advantage is accomplished with a decrease in the uncorrected distance visual acuity. PMID- 11488006 TI - [Regression analysis of idiopathic endothelial cell loss after perforating normal risk keratoplasty: basic principles for long-term analysis of endothelial risk factors in a retrospective clinical study]. AB - BACKGROUND: For the characterisation of influencing factors on chronic endothelial cell loss after penetrating keratoplasty by means of multivariate statistics, a mathematical description of the course of the individual postoperative endothelial cell density is a prerequisite. This mathematical description should result in a standardized index value describing course and amount of the postoperative endothelial cell loss over time in a canonical way. The slopes of the linear regression lines for each individual scatter plot of a) the endothelial cell density values plotted against the respective postoperative time directly (linear regression), and b) after logarithmic transformation (exponential regression) are evaluated, respectively. PATIENTS AND METHODS: 58 patients after normal-risk keratoplasty (26x keratokonus, 22x Fuchs-dystrophy and 10 cases of corneal decompensation after cataract surgery) with 5 or more postoperatively acquired endothelial density values and without any episodes of graft rejection were included in this study. Mean follow up was 2.9 +/- 1.1 years. The postoperative endothelial cell density values were plotted against the respective time for each patient individually. The coefficients of variation (R2) derived from the linear and the exponential regression models were calculated for each of these scatter plots. The pairs of R2 values (linear vs. exponential) were compared statistically. A dependence of the difference of linear and exponential R2-values on the ophthalmologic diagnosis was tested as well. RESULTS: The linear model is able to declare 83% the total variance of the course of the endothelial cell density. The exponential model even declares 86%. This small difference was statistically significant. Since both methods of regression describe the course of the cell density well, intra/and extrapolation of missing endothelial values is possible with both models. No dependence of the difference of linear and exponential R2-values on the ophthalmologic diagnosis could be demonstrated. CONCLUSIONS: Both, the intuitively understandable slope of the linear regression line and the constant of decay of the exponential regression curve, are suitable for describing the amount of the postoperative loss of endothelial cells after normal-risk keratoplasty independent of the ophthalmologic diagnosis. Both can thus be used as target variable in forthcoming statistical analyses for chronic endothelial cell loss. PMID- 11488007 TI - [Excimer laser versus diamond fraise: equal short-term outcome of corneal smoothing in pterygium operations]. AB - BACKGROUND: The treatment of pterygia often results in irregular astigmatism. Corneal smoothing at the end of surgery plays a major role in the development of astigmatism. The aim of our study was to compare the efficiency of corneal smoothing performed by phototherapeutic keratectomy compared with the conventional method performed by a diamond fraise. The effect of postoperative excimer laser smoothing on refraction, visual acuity and irregular astigmatism was investigated. PATIENTS AND METHODS: 32 eyes underwent pterygium excision using a standardized surgical procedure. In addition 9 eyes underwent postoperative phototherapeutic keratectomy of the wound region with an excimer laser, 23 eyes were treated with a diamond fraise for corneal smoothing. The postoperative examination was performed 7, 14 and 30 days after surgery. Corneal topography was performed by a computer-assisted videokeratoscope (TMS-1). The results were evaluated by the Fourier transform analysis. RESULTS: Both methods showed similar results concerning refraction and visual acuity postoperatively. Preoperatively all patients had an irregular flattening of the corneal topography in the area of the pterygium. Postoperatively a small irregular flattening was left in eyes treated with a diamond fraise. In the excimer laser treated group a steepening in the corneal topography could be observed. CONCLUSIONS: The phototherapeutic keratectomy with excimer laser in the treatment of pterygia suggests to be a gentle alternative creating a smooth wound surface. Despite of this advantage the treatment with the excimer laser effects a remarkable increase in administration and expense. PMID- 11488008 TI - [Histopathology of 8 corneal buttons after penetrating keratoplasty in silicone oil-associated keratopathy]. AB - BACKGROUND: In the literature there is only a limited number of morphological reports on clinically diagnosed silicone oil associated keratopathy describing different histological changes. The purpose of the present study was to evaluate the most common histopathological features of this disorder. MATERIAL AND METHODS: We reviewed the registry of the ophthalmopathological laboratory at the University of Marburg with respect to the following histopathological diagnoses: "bullous keratopathy", "endothelial-epithelial corneal decompensation", "band keratopathy" and "endothelial degeneration". These specimens were cross-checked with appropriate medical records. Eight specimens with a clinical diagnosis of silicone oil induced keratopathy were identified. All specimens were examined by light microscopy. RESULTS: Histologically, a long-standing bullous keratopathy was seen in 5 out of 8 specimens. A descemetocele was present in two other corneas. One case displayed band keratopathy. A posterior collagenous layer (PCL) between Descemet's membrane and the endothelium was identified in 7 out of 8 specimens examined. This layer was of a fibrillar type in 2 corneal buttons and of a fibrocellular type in all remaining specimens with PCL. PCL was associated with endothelial cell loss and degeneration. The endothelial cell density varied between 0 and 5 cells per high power field. CONCLUSION: Posterior collagenous layer associated with degenerating endothelium appears to be the most frequent histopathological feature in silicone oil induced keratopathy. The variety of PCL in this condition makes a firm histopathological diagnosis of "Silicone oil induced keratopathy" rather difficult. PMID- 11488009 TI - [Treatment of 9 squamous epithelial carcinoma in situ lesions of the conjunctiva (CIN) with mitomycin C eyedrops in cytological and DNA image cytometric control]. AB - BACKGROUND: Conjunctival intraepithelial neoplasia (CIN) is a frequent conjunctival tumor. Following excision alone recurrences are frequent. An effective postsurgical recurrence prevention is therefore highly desirable. In this study we aimed to evaluate the effectivity of postsurgical chemotherapy of conjunctival squamous cell carcinoma in situ (CIN) with mitomycin C eyedrops. We introduced the otherwise established diagnostic tools of cytology and DNA-image cytometry to the diagnosis and therapy-monitoring of CIN. PATIENTS AND METHODS: We treated 9 patients with CIN. For diagnosis the results of cytology and cytometry of presurgically obtained brush smears were compared with the histologic evaluation of the excised tissue. After surgery, we administered topical chemotherapy with mitomycin C eye drops 0.02% (MMC). Conjunctival brush smears were again evaluated by cytology and DNA-image-cytometry for postsurgical therapy monitoring. RESULTS: The clinical diagnosis of CIN was fully confirmed by cytology, DNA-image-cytometry and histology respectively in 7 patients. In one patient, the results of the applied diagnostic methods differed in results: Histologic evaluation indicated a moderate dysplasia but DNA-image-cytometry showed significant DNA-aneuploidy unequivocally indicating neoplasia like squamous cell carcinoma. In another patient the preoperatively obtained conjunctival brush smears could not properly analyzed by cytometry but clinical diagnosis was confirmed by histology. MMC-therapy was well tolerated except for a self-limited conjunctivitis. A complete remission of CIN was obtained in 8 of 9 patients (89%) who were free of CIN recurrences during a follow-up period of 27.2 months (11-48). Only one patient suffered from a recurrence 14 months after surgery and after 2 MMC-cycles. CONCLUSION: Adjuvant topical mitomycin C appears to be effective in the prevention of recurrences of conjunctival CIN after surgical removal. Our results indicate that at least 4 cycles of topical MMC are required to prevent local recurrences in the long term. Cytology and DNA-image cytometry are highly sensitive and specific methods for diagnosis and therapy monitoring of conjunctival squamous cell carcinoma in situ (CIN). PMID- 11488011 TI - [Accuracy of measurements of video-refractometry in higher ametropia]. AB - BACKGROUND: Measurement of refraction of non-cooperative patients is a real challenge for the ophthalmologist. Retinoscopy produces exact results in experienced hands, but requires practice and time. The Videorefractor VRB 200 (manufactured by Tomey, Erlangen) was especially designed for refracting children. The aim of this study is to prove the accuracy of this tool in measuring refraction. MATERIAL AND METHODS: 259 patients (age 10 month to 77 years) were measured with the VRB 200. Each patient was examined by retinoscopy in cycloplegia and refracted with the Topcon autorefraktor RM-A 6000. Ametropia was evoked at 5 emmetropic volunteers by lenses of different power, placed in a trial frame. These measurements were performed with and without use of cycloplegic drugs. RESULTS: The VRB 200 showed an average error of spherical equivalent at ametropia of the range -4 to +4 Diopters (dpt) < 2 dpt. At higher ametropia the error grew increasingly. Repeated measurements of one patient showed high variation of the results. Astigmatism was measured worse by growing magnitude, especially at diagonal axis (45 degrees and 135 degrees). Anisometropia > 2 dpt has been discovered in only 72%. It was possible to examine many infants and handicapped patients, which was impossible by using the RM-A 6000. An examination by retinoscopy was possible in all cases, but needed a lot of time and patience. CONCLUSIONS: The concept of good handling suffers especially by the increasing inaccuracy at ametropia beyond +/- 4 dpt. At this state of development the VRB 200 is no alternative in practical use to the conventional retinoscopy. PMID- 11488010 TI - [The closed Landolt ring--a handy test for evaluating suspected simulation]. AB - PURPOSE: We present a visual acuity test for proof of malingering and psychogenic impairment of visual acuity. METHOD: The book contains 36 plates. On 32 plates, a Landolt-C is shown with a gap of 10 minutes of arc, thus corresponding to a visual acuity of 0.1, when applied at a distance of 1 metre. Each of the 4 alternatives of the Landolt-C occurs eight times in random order. Starting at position 21, four plates are interspersed showing a closed circle of the same size. The test is used at a distance corresponding to an acuity level of maximum 50% of the presumed factual acuity. Following a four alternative forced choice paradigm, the tested person is requested to call the direction of the Landolt-C within about 2 seconds, when the plates of the book are turned over. The responses and their latency are recorded using a digital electronic system which allows later audiovisual analysis. The response on the first closed circle and its latency is compared to the previous responses. Two subjects were tested who tried to pretend a reduction of their visual acuity. RESULTS: Malingering was proved by the verbal response of the first subject and by the long latency of the response of the second subject (4.0 s vs. 1.2 s-2.7 s for the previous responses). The entire test, explanation included, took less than 5 minutes. CONCLUSION: This handy test can be a useful tool in cases suspect of malingering or psychogenic impairment of visual acuity. PMID- 11488012 TI - Elevated concentration of intraocular cortisol obtained by sequential parabulbar injection in rabbit model. AB - BACKGROUND: Parabulbar injections of corticosteroids yield a high intraocular concentration, but the half-life is short. We determined the concentration of cortisol that might be obtained by sequential parabulbar infusions of hydrocortisone. MATERIALS AND METHODS: Single parabulbar injections of hydrocortisone sodium succinate were made in a rabbit model and the time of maximum concentration and half-life determined in the retina-choroid, vitreous, serum, and the tissues of the paired eye by radio-immune assay. Based upon the half-life, sequential injections were made at 3 hour intervals to obtain a steady concentration. RESULTS: The maximum concentration of cortisol in the ocular tissues occurred two hours after a single injection and was 98 times greater in the choroid-retina and 72 times greater in the optic nerve than in the paired eye and the serum. The half-life of cortisol was 15 minutes. Elevated levels could be sustained by sequential injections at 3 hour intervals. CONCLUSION: Sequential injections of hydrocortisone through a retained parabulbar catheter could make it possible to titrate and maintain a therapeutic level of cortisol in the eyes of patients with posterior uveitis or optic neuritis. PMID- 11488013 TI - [Enucleation as a form of self-aggression--2 case reports and review of the literature]. AB - BACKGROUND: Autoenucleation is the worst form of self-mutilation of the eye. Patients affected are mostly those with psychosis and delusions, but such acts can also happen under the influence of drugs, alcohol, solvents and seldom with organic illnesses. Often, these patients are known to be autoaggressive and suicidal. PATIENTS: We present two psychiatric patients who autoenucleated one eye and injured the other. A 41-year-old patient with known organic epilepsy based on a temporal-lobe-hemangioma and psychosis with delusions autoenucleated his right eye and lacerated the conjunctiva of the other, following imperative voices. A 50-year-old female suffered from a long-standing paranoid schizophrenia and religious delusions, and autoaggressive acts with resulting amaurosis of the right eye and several suicidal attempts had preceded the autoenucleation of the left eye. DISCUSSION: After the completion of the autoaggressive act the patients experience relief, but often the autoaggresive or suicidal impulses persist or recur. A close cooperation between ophthalmologists and psychiatrist in these patients is imperative. PMID- 11488014 TI - [Zinsser-Engman-Cole syndrome (dyskeratosis congenita) with severe sicca syndrome, panuveitis and corneal perforation--a case report]. AB - BACKGROUND: Zinsser-Engman-Cole syndrome (Z.E.C.) is a very rare type of ectodermal dysplasia, inherited in X-linked recessive manner and characterised by poikiloderma, nail dystrophy, lingual leucoplakia, bone marrow hypoplasia, hyperkeratosis and hyperhidrosis of planta and palms, dental anomalies and caries, premature grey hair. PATIENT AND METHODS: We report on a 46-year-old man who presented with occlusion of lacrimal puncta, trichiasis, severe dry eye, recurrent corneal ulceration and perforation, uveitis. HLA typing, flow cytometry of peripheral lymphocytes, bone marrow biopsy, conjunctival biopsy and extensive laboratory evaluation towards autoimmune and infectious diseases were performed. RESULTS: CD4+ T cells fraction was decreased, CD8+ and CD3+ HLA DR+ elevated. The patient was HLA-B27 positive. Laboratory studies revealed increased erythrocyte sedimentation rate and C-reactive protein level, hypochromic and hypoplastic anaemia, negative serum titers of antibodies to Epstein-Barr virus, HIV, HTLV-I, toxoplasma gondii and treponema pallidum, repeated titers to cytomegalovirus, herpes simplex and herpes zoster viruses--IgM negative, IgG positive. Corneal perforation was treated with amniotic membrane transplantation and corneal transplantation. CONCLUSION: The defect in cell-mediated immune mechanisms in Z.E.C. syndrome explains the corneal perforation, sicca syndrome and uveitis, first reported in this syndrome. PMID- 11488015 TI - [Detection of hepatitis C virus RNA in tear film of a patient with recurrent peripheral corneal ulcers]. AB - BACKGROUND: There are conflicting reports on the role of hepatitis C virus in corneal pathology. PATIENT: A 58-year-old male patient presented with recurrent peripheral corneal ulcers and corneal thinning in the left eye. There was a bilateral vascular pannus formation and a decreased ocular wetting measured by Schirmer testing. The posterior ocular segment was normal. There was no sign of any systemic rheumatic disease. Serological testing detected antibodies against hepatitis C virus. Hepatitis C virus RNA testing using a quantitative polymerase chain reaction method revealed hepatitis C virus RNA in serum (> 3.2 million copies/ml) and in tear samples (18,000 copies/ml) of the patient. In a control group of 7 consecutive patients with hepatitis C virus RNA detection in the serum but without ocular pathology, no hepatitis C virus RNA was detected in tear samples (detection limit: 1000 copies/ml). CONCLUSIONS: Detection of hepatitis C virus RNA in lacrimal fluid of a patient with recurrent peripheral corneal ulcers may indicate a pathogenic role of hepatitis C virus in corneal pathology. Especially, since our patients with systemic hepatitis C virus infection but without ocular changes did not show hepatitis C virus RNA in their tears. Therefore, patients with recurrent corneal ulcers of unknown origin should be tested for systemic hepatitis C virus infection. PMID- 11488016 TI - [Conjunctival neuromas and prominent corneal nerve fibers as diagnostic indication of multiple endocrine disease]. AB - BACKGROUND: Multiple endocrine neoplasias (MEN-syndromes) represent diverse, mostly autosomal-dominant inherited, seldom sporadic diseases. MEN 2B-syndrome comprises medullary thyroid carcinoma, pheochromocytoma and mucosal neuromas. PATIENT: A 30 year old male patient presented with bilateral chronic irritation of the ocular surface. The biomicroscopy revealed intraconjunctival worm-like alterations, prominent corneal nerves and nodules on the upper lid margins. HISTOLOGY: The performed conjunctival biopsy showed nearly no goblet cells, but prominent intrastromal, immunohistochemical S100-positive neuromas. RESULTS: The initiated investigations revealed bilateral multifocal medullary thyroid carcinoma and a left sited pheochromocytoma. CONCLUSION: Conjunctival neuromas and prominent corneal nerves can be diagnostic for the MEN 2B-syndrom. Early diagnosis and prophylactic thyroidectomy are expected to improve the life expectancy even in asymptomatic gene-carriers. PMID- 11488017 TI - [Comment on Velten IM, Bergua A, Horm FK, Junemann A, Korth M. "Central corneal thickness in normal probands, patients with ocular hypertension, normal pressure and open-angle glaucomas--a clinical study"]. PMID- 11488018 TI - [In memoriam. Univ.-Prof. Hans Lauber (1876-1952)--"visual field Lauber"--was born 125 years ago]. PMID- 11488020 TI - [Anatomical study and clinical application of long head of triceps muscle for reconstruction of shoulder abduction]. AB - OBJECTIVE: To study the morphological character of long head of triceps muscle for clinical application in reconstruction of shoulder abduction. METHODS: Forty four upper extremities of fixed human adult cadavers were carefully dissected. The origins and the pedicles of blood vessels and nerves of long head of triceps muscle, as well as the maximum available size of the muscles, were measured. Six cases of clinical application of long head of triceps muscle for reconstruction of shoulder abduction were followed up for 3 to 11 months. RESULTS: The origins in the dorsal side of long head of triceps muscle were muscular and the ventral side were tendinous, which was 7.6 to 13.3 cm in length and 1.6 to 3.4 cm in width. The distance from the origin to the neurovascular pedicle was 5.7 to 11.4 cm. The radial nerve, which innervated the muscles, could be dissected for 2.9 to 11.8 cm in length. The blood supplies to the triceps muscle were from humeral artery (43.2%), 1.0 to 6.0 cm in length and 1.6 to 2.4 mm in diameter, and from humeral profundus artery (45.5%), 1.5 to 4.4 cm in length and 0.9 to 2.4 mm in diameter, if the vessel was separated to the humeral artery, the length was 1.5 to 6.3 cm. The neurovascular pedicles were multiple branched. In the 6 cases of clinical application of the triceps muscles, the operated shoulder could abduct from 5 degrees preoperatively (0 degree to 10 degrees) to 77.3 degrees (50 degrees to 90 degrees) postoperatively. CONCLUSION: In accordance to the anatomical character of the triceps muscles, the long head of triceps muscle is a suitable choice for reconstruction of shoulder abduction with optimistic outcomes. PMID- 11488021 TI - [Reconstruction of digital avulsion with pre-fabricated subdermal vascular network skin flap by ultrasonic liposuction]. AB - OBJECTIVE: To investigate the clinical application of subdermal vascular network skin flap pre-fabricated by ultrasonic liposuction in reconstruction of digital avulsion. METHODS: Forty-seven injured fingers of 23 cases were treated from June 1997 to February 2000. Conventional abdominal skin flap was elevated, according to the size of digital avulsion, and subcutaneous fat was removed with scissors. Ultrasonic liposuction technology was adopted, in order to minimize the injury of subdermal vascular vessels, to remove the fat particles close to the vascular network. Finally, the pre-fabricated skin flap was used to repair the digital avulsion. The vascular pedicle was severed in 5 to 7 days after operation. The range of skin flap was 4 cm x 3 cm to 8 cm x 7 cm, and the ratio of length and width was (2 to 3) to 1. RESULTS: All the skin flaps were survived. Twenty-one patients were available for postoperative follow-up for 6 to 24 months. The motion of interphalangeal joint achieved functional recovery, and the sensation of pain, temperature and taction recovered well. CONCLUSION: Ultrasonic liposuction does not obviously injure the subdermal vascular network skin flap, it is a simple and safe method for treatment of digital avulsion. PMID- 11488022 TI - [The diagnosis and treatment of quadrilateral space syndrome]. AB - OBJECTIVE: To investigate the compression feature, clinical manifestation and the results of treatment of quadrilateral space syndrome. METHODS: Four patients with axillary nerve entrapment at quadrilateral space had been treated and followed up for 5 to 12 months from May 1999 to June 2000. The causes, symptoms, signs and the treatment management of those cases were analyzed. RESULTS: Among the 3 cases which received operation, sensation and motor function completely recovered in 2 cases and partially recovered in 1 case. No obvious recovery of sensation and motor function in the case which received local nerve blocking treatment. CONCLUSION: The main diagnostic evidence for axillary nerve entrapment is the deltoid muscle paralysis and paresthesia in the lateral side of shoulder, and early neurolysis is recommended as soon as the diagnosis is clarified. PMID- 11488023 TI - [Experimental study in rabbit on repair of radial bone defect by bioactive degradable polymer]. AB - OBJECTIVE: To investigate the availability of bone defect repair by degradable porous polycaprolactam (PCL) as the carrier of bone morphogenetic protein (BMP). METHODS: Three different kinds of bone substitutes, including decalcified bone matrix, PCL-BMP compounds and simple PCL, were implanted into the radial bone defects in 36 rabbits, and in the other 12 rabbits the bone defects were left untreated as control. X-ray examination, X-ray morphometry, histological and electron microscopic observation were performed at different time after operation. RESULTS: The quantity of new bone formation in PCL-BMP group was prior to that in simple PCL group. The pattern and speed of bone defect repair in PCL BMP group were similar to those in decalcified bone matrix. Electron microscopic observation showed that the PCL-BMP group degraded faster than simple PCL group, which was more suitable for bone repair. CONCLUSION: PCL is a good carrier of BMP with potential for clinical use. PMID- 11488024 TI - [Study on transplantation of anterolateral femoral skin flap with high site direct cutaneous artery]. AB - OBJECTIVE: To study the availability of transplantation of anterolateral femoral skin flap with absence of the first branch of musculocutaneous artery. METHODS: One hundred and sixty cases of anterolateral femoral skin flap transplantation in the past 15 years were reviewed, to investigate the relationship between the outcome of the transplantation and the types, distribution, original point, diameter of blood vessels. Among the 160 cases, there were 10 cases, 6.3% in total, of skin flap transplantation with the pedicle of high site direct cutaneous femoral artery, ranging from 10 cm x 14 cm to 12 cm x 18 cm in size, applied to repair soft tissue defect. RESULTS: There were 149 cases, among the 160 cases, survived and all the 10 cases of femoral skin flap transplantation with high site anterolateral cutaneous artery survived. CONCLUSION: The femoral skin flap with high site anterolateral cutaneous artery has good blood supply, larger size available, and easier operation. When the first branch of descending anterolateral femoral artery is absent, the femoral skin flap with high site anterolateral cutaneous artery is an optimal alternative. PMID- 11488025 TI - [The anatomic study and clinical application of the bone (periosteum) flap pedicled with upper muscular branches of lateral femoral muscle]. AB - OBJECTIVE: To explore the anatomic feature and clinical application of the bone (periosteum) flap pedicled with upper muscular branches of lateral femoral muscle. METHODS: The anatomic features and distribution of upper muscular branches of lateral femoral muscle were observed in the lower extremities of 40 adult cadavers. From February 1989 to February 1999, 7 cases with bone defect or nonunion of upper part of femur were treated with transfer of bone (periosteum) flap pedicled with upper muscular branches of lateral femoral muscle. RESULTS: The upper muscular branches of lateral femoral muscle originated from the transversal branch of lateral circumflex femoral artery. The musculoperiosteal branch and periosteal branch were originated at 16.8 +/- 3.0 cm below the greater trochanter. The diameter and length of musculoperiosteal branch were 1.4 to 1.7 mm and 2.7 to 5.6 cm, those of the periosteal branch were 0.4 to 0.6 mm and 1.2 to 1.5 cm respectively. Bone union achieved in 10 to 18 weeks after operation in all 7 cases after 18 to 42 months follow-up. The motion of hip joint reached 180 degrees in 4 cases, 120 degrees in 2 cases and 65 degrees in 1 case. The donor area recovered well. CONCLUSION: The bone (periosteum) flap pedicled with upper muscular branches of lateral femoral muscle is an effective alternative for repairing the bone defect or nonunion of the upper or middle part of femur. PMID- 11488026 TI - [Reconstruction of heel by reversed island fibular musculocutaneous flap]. AB - OBJECTIVE: To explore the anatomical basis of blood supply and heel reconstruction by reversed island fibular musculocutaneous flap. METHODS: The blood supply of fibular musculocutaneous flap and the biomechanical characteristics of heel were studied by anatomical examination. One case with right heel full defect because of explosion injury was repaired by transfer of reversed island fibular vessels. The fibular flap was 14 cm in length with part of peroneus muscle and long flexor muscle of great toe. RESULTS: The lower part of fibular artery had plentiful anastomosis with anterior tibial artery and posterior tibial artery, which could provide ideal reversed blood supply. The rotatory point of vessel pedicle could be chosen according to the need of operation. The lowest site might be above 6 cm to lateral malleolus, and the vessel pedicle was 20 cm in length. The morphological feature of the reversed island fibular musculocutaneous flap was suitable to the biomechanical character of heel. The patient achieved satisfactory clinical result, the musculocutaneous flap survived well for 10 months of follow-up. CONCLUSION: The reversed island fibular musculocutaneous flap provide a new method for repairing the severe heel defect, especially in full defect of calcaneus and cuboid bone. PMID- 11488027 TI - [The therapy effect of improved bilateral tibia lengthening]. AB - OBJECTIVE: To study the therapy effect of improved bilateral tibia lengthening. METHODS: From May 1997 to May 2000, 32 patients (varus knee deformity in 8 cases) with low stature were adopted in this study. Among them, there were 26 females and 6 males, aged from 18 to 45 years old. Operative procedures included: 1. tibia osteotomy 1 cm distal from tibia tuberosity and fibula osteotomy 10 cm proximal from lateral malleolus; 2. fixation of the tibia osteotomy with interlocking nail and locking the proximal nail; 3. fixation of the lengthening apparatus; 4. lengthened bilateral tibia 0.7 mm per day; 5. removed the apparatus and locked the distal nail 2 weeks later after limb lengthening was over. RESULTS: The mean distance of lengthening was 8.5 cm (ranged 3.5 to 12.0 cm), the mean duration of lengthening was 128 days(ranged 53 to 180 days), and the mean time of bone union was 180 days (ranged 120 to 270 days). Followed up for 1 to 3 years, 98% patients felt satisfactory in lengthening, gait and joint movement. CONCLUSION: The improved bilateral tibia lengthening technique is recommended for advantage of short time of bone union, less complication and correcting the varus deformity of knee simultaneously. PMID- 11488028 TI - [The clinical application of distally based neurocutaneous flaps by anastomosis of superficial veins]. AB - OBJECTIVE: To investigate the clinical results of the distally based neurocutaneous flap by anastomosis of superficial veins. METHODS: From June 1996, 19 cases with composite skin defects of the distal part of limb were repaired by the transposition of distally based neurocutaneous flaps, including traumatic defect in 10 cases, chronic ulcer in 3 cases, scar contracture in 6 cases. The distally based sural neurocutaneous flaps were used in 9 cases, the reverse-flow saphenous neurocutaneous island flaps were used in 2 cases, and the retrograde neurocutaneous island flaps of the forearm were used in 8 cases. The flap area ranged from 15 x 24 cm to 4 x 6 cm, the pedicle of the flap ranged from 6 cm to 15 cm in length. The superficial vein of the flap were anastomosed with the subcutaneous superficial vein of the recipient site to improve the venous drainage. RESULTS: The composite flap survived completely in 17 cases. One cases with retrograde-flow forearm neurocutaneous flap and another case with reversed sural neurocutaneous flap were partially survived because of thrombosis in anastomosed veins postoperatively. Sixteen cases were followed-up for 6 to 24 months, the color and texture of the flap were excellent, the protective sensation were recovered, the configuration and function were satisfactory. CONCLUSION: Anastomosis of superficial veins of the composite flaps with the subcutaneous superficial veins of the recipient site can significantly improve the venous drainage, enlarge the survival area of the flap and the reparable area. PMID- 11488029 TI - [Treatment of the pyogenic infection of the thoracolumbar intervertebral space with vascular pedicled rib grafting]. AB - OBJECTIVE: To investigate the effect of vascular pedicled rib grafting in treatment of the pyogenic infection of the thoracolumbar intervertebral space. METHODS: From November 1993, 7 cases with pyogenic infection of the thoracolumbar intervertebral space were adopted in this study. Among them, there were 5 males and 2 females, aged from 29 to 58 years old. The position of pyogenic infection located at T10 to 11 in 3 cases, at T11 to 12 in 2 cases and at T12 to L1 in 2 cases. During operation, the pyogenic infection was cleared completely and the rib with vascular pedicle was grafted into the intervertebral space. RESULTS: Followed up for 10 to 60 months, the bone graft unioned in 3 months for 5 patients and unioned in 4 months for 2 patients. There was no recurrence and raised kyphosis. CONCLUSION: The technique of vascular pedicled rib graft is simple, it can shorten the bone union time and reconstruct the stability of spine. PMID- 11488030 TI - [Initiative report of the hydroxyapatite orbital implant drilling procedure]. AB - OBJECTIVE: To evaluate the clinical effect of drilling procedure following the hydroxyapatite orbital implantation. METHODS: From February 1996 to April 2000, 146 consecutive patients who received hydroxyapatite orbital implant were drilled and inserted a motility peg 6 to 16 months after hydroxyapatite implantation. Among them, there were 97 males and 49 females, aged from 18 to 60 years old, of the 146 motility pegs, 36 were sleeved pegs and 110 were nonsleeved. Goldman visual field analyzer was applied to measure the degree of artificial eye's movement before and after drilling. RESULTS: Followed up for 1 to 40 months, no secondary infection occurred. The mobility of the prosthesis increased from (18.7 +/- 3.8) degrees preoperatively to (42.3 +/- 3.7) degrees postoperatively. CONCLUSION: The delayed drilling procedure and motility peg insertion improve the range of movement and the sensitivity of the artificial eye with a low rate of complications. PMID- 11488031 TI - [Study on the molecular mechanisms involved in the increased collagen synthesis by platelet-derived wound healing factors during wound healing in alloxan-induced diabetic rat]. AB - OBJECTIVE: To explore the molecular mechanisms involved in the increased collagen synthesis by platelet-derived wound healing factors (PDWHF) during wound healing in alloxan-induced diabetic rats. METHODS: Thirty-three male SD rats were divided into two groups, the normal (n = 9) (group A) and the diabetic group (n = 24). Two pieces of full-thickness skin with diameter of 1.8 cm were removed from the dorsal site of diabetic rats. PDWHF (100 micrograms/wound) was topically applied to one side of the diabetic wounds (group B) on the operation day and then once a day in the next successive 6 days. Meanwhile, bovine serum albumin (100 micrograms/wound) was applied to the other side of diabetic wound as control group (group C) in the same way. Levels of transforming growth factor-beta 1 (TGF beta 1) and procollagen I mRNA in wound tissue were inspected by dot blotting. RESULTS: TGF-beta 1 mRNA levels in group B were 4 folds and 5.6 folds compared with those in group C after 5 and 7 days (P < 0.01), however, still significantly lower than those of group A (P < 0.05). There was no significance difference among three groups on the 10th day after wounding. The levels for procollagen I mRNA in group B amounted to 2.1, 1.8 and 2.3 folds of those in group C after 5, 7, and 10 days (P < 0.01), respectively. Compared with those in the group A, procollagen I mRNA levels in the group B were significantly lower after 5 and 7 days (P < 0.05), and no significant difference was observed between group B and A after 10 days. CONCLUSION: One important way for PDWHF to enhance the collagen synthesis in diabetic wound healing is to increase the gene expression of endogenous TGF-beta 1. PMID- 11488032 TI - [Cellular compatibility of three natural xenogeneic bone derived biomaterials]. AB - OBJECTIVE: To evaluate the cellular compatibility of three natural xenogeneic bone derived biomaterials. METHODS: Three types of natural xenogeneic bone derived biomaterials were made with physical and chemical treatment, composite fully deproteinized bone(CFDB), partially deproteinized bone(PDPB) and partially decalcified bone(PDCB). Three types biomaterials were cocultured with human embryonic periosteal osteoblasts. The cell growth, attachment, cell cycle, alkaline phosphatase activity were detected to evaluate the cellular compatibility to biomaterials. RESULTS: Osteoblasts attached on all three biomaterials and grew well, the effect of three biomaterials on cell proliferation was PDCB > PDPB > CFDB. The cell cycle was not obviously affected by three biomaterials. The effect of three biomaterials on alkaline phosphatase activity of osteoblasts was PDCB > PDPB > CFDB. CONCLUSION: CFDB,PDPB,PDCB have good cellular compatibility without cytotoxic and tumorigenicity, CFDB is the best. The three biomaterials can be used as scaffold materials of bone tissue engineering. PMID- 11488033 TI - [The effects of dexamethasone on biological characteristics of bone marrow stromal cells]. AB - OBJECTIVE: To investigate the effects of dexamethasone on the proliferation and differentiation of bone marrow stromal cells(MSC). METHODS: MSC were isolated and cultured in vitro. After treatment with different concentrations of dexamethasone (0, 10(-10), 10(-9), 10(-8), 10(-7) and 10(-6) mol/L), the proliferation and alkaline phosphatase (ALP) activity of MSC were measured to evaluate the effect of dexamethasone on the biological characteristics of MSC. RESULTS: Dexamethasone inhibited cell proliferation. With the increase of concentration of dexamethasone, the effect was enhanced, which was more significant when the concentration of dexamethasone was over 10(-8) mol/L. At the same time, dexamethasone promoted the activity of ALP. This effect was enhanced with the increase of concentration of dexamethasone, but the alteration was small when the concentration of dexamethasone was over 10(-8) mol/L. The effects increased with the time. The activity of ALP was enhanced 2 to 4 times with the dexamethasone for 6 days. CONCLUSION: Dexamethasone inhabit the proliferation of MSC, while induce them to differentiate into osteoblasts. The appropriate concentration of dexamethasone was 10(-8) mol/L. PMID- 11488034 TI - [Fabrication of tissue engineered skin equivalent]. AB - OBJECTIVE: To fabricate artificial human skin with the tissue engineering methods. METHODS: The artificial epidermis and dermis were fabricated based on the successful achievements of culturing human keratinocytes(Kc) and fibroblasts (Fb) as well as fabrication of collagen lattice. It included: 1. Culture of epidermal keratinocytes and dermal fibroblasts: Kc isolated from adult foreskin by digestion of trypsin-dispase. Followed by comparison from aspects of proliferation, differentiation of the Kc, overgrowth of Fb and cost-benefits. 2. Fabrication of extracellular matrix sponge: collagen was extracted from skin by limited pepsin digestion, purified with primary and step salt fraction, and identified by SDS-PAGE. The matrix lattice was fabricated by freeze-dryer and cross-linked with glutaraldehyde, in which the collagen appeared white, fibrous, connected and formed pores with average dimension of 180 to 260 microns. 3. Fabrication artificial human skin: The artificial skin was fabricated by plating subcultured Kc and Fb separately into the lattice with certain cell density, cultured for one week or so under culture medium, then changed to air-liquid interface, and cultured for intervals. RESULTS: The artificial skin was composed of dermis and epidermis under light microscope. Epidermis of the skin consisted of Kc at various proliferation and differentiation stages, which proliferated and differentiated into basal cell layer, prickle cell layer, granular layer, and cornified layer. Conifilament not only increased in number, but also gathered into bundles. Keratohyalin granules at different development stages increased and became typical. The kinetic process of biochemistry of the skin was coincide with the changes on morphology. CONCLUSION: Tissue engineered skin equivalent has potential prospects in application of repairing skin defect with advantages of safe, effective and practical alternatives. PMID- 11488035 TI - [The effect of Chinese cobra venom factor on guinea pig to rat cardiac xenotransplantation]. AB - OBJECTIVE: In the guinea pig-to-rat cardiac xenotransplantation model, the effect of complement depletion by using Chinese Cobra Venom Factor(CVF) on hyperacute rejection was evaluated. METHODS: Cardiac xenograft from guinea pig was transplanted into the abdomen of rat after the recipient being given i.p. a dose of CVF 0.20 microgram/g. the recipients were divided into group A (control group), group B (only given CVF), group C (CVF + Cytoxan + Splenectomy), group D (Cytoxan + Splenectomy) Cytoxan was injected into the abdominal cavity with a dose of 60 mg/Kg. The survival time of xenograft was measured and histologic observation was carried out after the cardiac arrest. RESULTS: The survival time of xenograft ranged from 15 to 3,120 minutes. There were significant difference among group A compared with group B and C (P < 0.01), and no difference between group A and group D, as well as group B and C (P > 0.05). There were significant difference between group B and D, as well as group C and D(P < 0.01). The histologic observation proved that the hyperacute rejection in group A and D was milder than group B and C. CONCLUSION: The study reveals that CVF can prolong the xenograft time by depleting complement activities and restricting hyperacute rejection in this model. Further basic and clinical study of effect of CVF in xenograft transplantation is worthwhile. PMID- 11488036 TI - [Allografting for massive bone defect: bone and soft tissue reconstruction and postoperative rehabilitation]. AB - OBJECTIVE: To evaluate the results of limb function and the methods of bone and soft tissue reconstruction of patients treated with allografting. METHODS: From May 1992 to January 1999, 90 patients suffered from bone malignant tumor were treated with allografting in different methods of internal fixations. The average follow-up was 37.5 months. The limb postoperative function, complications related to different surgical methods were compared according to Enneking evaluation system. RESULTS: Skin necrosis, infection, non-union, fracture of allograft were the main complications which affect patients' limb postoperative functions. Of the 90 fresh-frozen allografting procedures, the final results of operation showed that hip joints and knee joints were better than the shoulder joints. More than 80% of the patients treated with interlocked intramedullary nail and allograft-prosthesis combination led to an over-all result that was excellent and good. Interlocked intermedullary nail was of recommended method of internal fixation. Early exercises of operative limbs could promote function recovery. CONCLUSION: Using of interlocked intramedullary nail and allograft-prosthesis combination are of recommended operation method and can be applied with better results, and early exercises of operative limbs will lead to better functions. PMID- 11488037 TI - [The biological effect of matrix metalloproteinase-1 in epidermal repair]. AB - OBJECTIVE: To review the role of matrix metalloproteinase-1 (MMP-1) in the course of healing in wounded skin. METHODS: The recent literatures on MMP-1 in skin wound repair were reviewed, which gave the insight into the local effect of MMP-1 during re-epithelialization. RESULTS: Following injury, basal keratinocytes, moving from the wound edge and interact with dermal matrix proteins in the wound bed, were induced to express MMP-1 in a specific space-time pattern. MMP-1 cleaved the collagen, thereby altering its structure and affinity by which the keratinocytes binded it. MMP-1 served a beneficial role in wound healing by facilitating the proliferation and movement of keratinocytes over the collagen rich wound bed during re-epithelialization. CONCLUSION: MMP-1 expression of migrating keratinocytes directly influences the re-epithelialization during the course of healing of the wounded skin. PMID- 11488038 TI - Radiographic interpretation of normal skeletal variations and pseudolesions in the equine foot. AB - Effective radiographic interpretation requires a veterinarian who is knowledgeable of equine limb anatomy and the various principles that affect the resulting image. The normal and its variations must be recognized and understood before the abnormal can be confidently identified as pathologic. Proper patient positioning and sound radiographic technique are mandatory if reliable diagnostic radiographs are to be produced. This review emphasizes equine foot radiographic variations of normal and pseudolesions that occur with commonly used radiographic views performed in equine practice. PMID- 11488039 TI - Physical principles and technical considerations for equine computed tomography and magnetic resonance imaging. AB - This article discusses how cross-sectional imaging methods such as computed tomography and magnetic resonance imaging can provide unique and diagnostically important information in situations where radiography or diagnostic ultrasound have been unrewarding. PMID- 11488040 TI - Computed tomography and magnetic resonance imaging of the equine head. AB - Horses are frequently presented with clinical conditions warranting diagnostic imaging of the head. Unfortunately, evaluation of the equine head can be difficult using conventional radiography. The anatomic complexity and superimposition of the osseous, dental, and soft tissue structures complicate radiographic interpretation and diagnoses. Multiple radiographic projections may be necessary to image specific regions, and side-to-side comparison is often essential. PMID- 11488041 TI - Computed tomography and magnetic resonance imaging of the equine musculoskeletal conditions. AB - There is a growing interest in the use of computed tomography (CT) and magnetic resonance (MR) imaging in equine orthopedic patients. It is well established that CT and MR imaging offer superior diagnostic information in a wide variety of musculoskeletal injuries in human beings and small animals. The highly detailed cross-sectional images obtained with these two modalities can often demonstrate pathologic changes undetected with other common imaging techniques. Based on their multiple applications in human beings and small animals, CT or MR imaging may prove to be the optimal diagnostic imaging technique for several types of musculoskeletal disorders of horses. PMID- 11488042 TI - Superficial digital flexor tendon healing: ultrasonographic evaluation of therapies. AB - Until recently, it was difficult to critically evaluate tendon healing in vivo. Superficial digital flexor tendon injuries were considered healed when the injured tendon was cold, non-painful, adequate time had passed for tendon healing to occur, and no recurrence of the injury was detected when the horse returned to athletic work. This article discusses how ultrasonography has revolutionized the diagnosis, treatment, and management of tendon injuries. PMID- 11488043 TI - Stress echocardiography and its role in performance assessment. AB - Stress echocardiography in the horse is in its infancy, and investigations need to continue to determine the significance and mechanism of postexercise echocardiographic abnormalities detected in horses. This work should be coupled with investigations on the exercise-induced changes in cardiac isoenzymes, cTnI, arterial oxygen content, and cardiac rhythm that may occur concurrently. With continued investigations into the cardiovascular problems that occur in the exercising horse, the sport horse veterinarian should be better able to accurately diagnose and treat problems causing poor performance in the horse. PMID- 11488044 TI - Abdominal and thoracic radiography in the neonate. AB - Plain and contrast radiographic techniques are used infrequently in the diagnostic evaluation of abdominal or thoracic disease in the adult horse because of the animal's large body size and the limited availability and expense of suitable equipment. The importance of radiography as a critical tool has also been lessened through the accessibility of ultrasound, although this technique is limited by depth and offers only a superficial window. Traditional radiographic techniques can be readily used in young foals because of their small body size, and radiography frequently provides critical information that aids in decision making. The ability to physically restrain a foal allows not only standing lateral images, but laterally recumbent and ventrodorsal views without the necessity for general anesthesia. PMID- 11488045 TI - Computed radiography. AB - CR is a useful tool for the equine veterinarian and has many advantages. Its only major disadvantage is the initial cost, in dollars and time, for setup in one's practice. CR is already in use at several universities and private practices in the United States and around the world. In the future, as is the case with any computer-based technology, this modality should become more affordable and readily available to smaller practices. The potential of CR in veterinary medicine combined with saturation of the human market is driving the development of specialized software and algorithms for veterinary medicine. PMID- 11488046 TI - Equine nuclear scintigraphy. Indications, uses, and techniques. AB - The demand for advanced and correlative imaging methods in equine medicine is growing. Scintigraphy can provide unique information about the physiologic status of an animal that cannot be discerned by other imaging modalities. PMID- 11488047 TI - Equine bone scintigraphic uptake patterns related to age, breed, and occupation. AB - It is well known that skeletal scintigraphy is an imaging modality with high sensitivity but low specificity. Sites of IRU provide good evidence of increased bone turnover but not its specific cause. Results of lameness workups, blocks, and other imaging techniques are required to determine specific causes. Overinterpretation of causes of IRU is tempting, especially in assigning diagnoses to EIBR--adaptive or non-adaptive. Nevertheless, a quarter of a century's experience with equine skeletal scintigraphy has shown that certain patterns and locations can be fairly predictive of certain pathologic findings. It behooves the equine scintigrapher to know those patterns and how they are affected by the patient's age, breed, and occupation so that he or she can indicate how to best focus other diagnostic procedures toward a definitive diagnosis. Those who are conducting research involving equine skeletal scintigraphy are challenged to help sharpen that focus, especially in evaluating the significance of relatively mild IRUs. Equine clinicians who use scintigraphy in their lameness workups are especially challenged. Working in concert with nuclear medicine imagers, they are in the best position to garner, collate, and analyze the information that is needed to correlate IRUs and the clinical picture. As difficult as they are to design and conduct, such studies are most productive if done prospectively and are most rewarding for those who carry them through. PMID- 11488048 TI - Diagnostic thermography. AB - Thermography is a practical aid in the clinical evaluation of the equine patient. It is particularly germane to the evaluation of lameness. This modality specifically increases the accuracy of diagnosis. Thermography is the pictorial representation of skin temperature. The technique involves the detection of infrared radiation, which can be directly correlated to blood flow. To be accurate, thermography must be performed in a controlled area free of drafts. The area should be protected from sunlight to avoid erroneous heating of the skin, and the horse's hair length should be uniform. Thermography detects heat before it is perceptible during routine physical examination and thus is useful for the early detection of laminitis, stress fractures, and tendinitis. It offers a noninvasive means of evaluating the blood supply to an injured region and represents one of the only reliable noninvasive means to evaluate blood flow to the foot of the horse. Thermography is also useful for the early identification of stress injuries to the contralateral limb of convalescing orthopedic patients. Thermography is an excellent adjunct to clinical examination as well as being complementary to other imaging techniques such as radiology, ultrasonography, and scintigraphy. PMID- 11488049 TI - Pedorthic management of the diabetic foot. AB - Foot ulceration in diabetic patients is a resource-consuming, disabling morbidity that often is the first step in the downward spiral to lower extremity amputation. The best treatment is prevention. Prevention of foot ulcers requires a coordinated program of foot-specific patient education, prophylactic skin and nail care, and protective footwear. The goal of prescription footwear is to keep individuals ambulatory, while protecting them from ulcer formation. The complexity of pedorthic prescription increases with decreasing protective sensation and increasing structural deformity of the foot and ankle. Prescription footwear accommodates deformity, while decreasing pressure and shear forces applied to skin overlying bone prominences. PMID- 11488050 TI - Orthotic and prosthetic devices in partial foot amputations. AB - With advanced surgical techniques and orthotic, as well as prosthetic devices, partial foot amputations have become a viable alternative. Orthotics can help restore stability, maintain support, and protect function of the residual limb. The authors discuss orthotic and prosthetic management of patients who have undergone toe amputations; ray amputations; transmetatarsal, Lisfranc-, or Chopart-level amputations. PMID- 11488051 TI - Inpatient management of lower extremity amputations. AB - The in-hospital care of the amputee is crucial; careful preoperative, intraoperative, and postoperative regimens set the stage for a successful clinical outcome. Caring for the patient undergoing amputation is much more than choosing the correct level at which to cut. Early aggressive rehabilitation; careful attention to postoperative complications; and appropriate physical therapy, occupational therapy, and counseling all play pivotal roles. Thorough knowledge of these issues helps to increase the likelihood of a successful outcome and to keep morbidity to a minimum. PMID- 11488052 TI - Orthopedic footwear. Custom-made and commercially manufactured footwear. AB - Shoes and footwear have been a part of human lives for centuries. Many take good foot health for granted. For those individuals, the commercial shoe store has hundreds of varieties of shoes in every price range. For individuals with compromised, at-risk feet, the options are reduced greatly. The foot health industry today has made available many viable options to meet specific needs. Footwear can augment successfully the treatment of foot and ankle pathology and when properly applied improve the health and lifestyle of patients. PMID- 11488053 TI - Orthotic devices. Custom-made, prefabricated, and material selection. AB - Orthotic use has become a common adjunct to medical treatment for foot and ankle pathology. For pain and discomfort from overuse to pathology-specific problems, interventions involving foot orthoses are widely accepted. A wide range of accommodative, functional, and prefabricated orthoses is available. With the emergence of technology in the manufacture of raw materials and prefabricated orthoses and the training and expertise of technicians and practitioners, the quality and accuracy of foot orthoses have improved vastly, justifying their role in the treatment and rehabilitation of patients with simple and complex foot problems. PMID- 11488054 TI - Optimal surgical preparation of the residual limb for prosthetic fitting in below knee amputations. AB - A well-performed below-knee amputation can result in a high level of patient satisfaction and function. The potential for a successful result and independent ambulation is good after a transtibial amputation, with approximately 70% to 100% of patients being able to ambulate independently after this procedure. A properly fitted and appropriately designed prosthesis is integral to the achievement of these goals. The approaches and techniques discussed here are essential in optimally preparing the residual limb for prosthetic fitting. Attention to factors such as the preoperative evaluation and the surgical technique can ensure the best possible outcome for amputation patients. PMID- 11488055 TI - Checklist for evaluating a transtibial prosthesis. AB - A thorough initial evaluation and ongoing follow-up are critical to an amputee's success with his or her prosthesis. A checklist is presented as a guide for the rehabilitation team to perform a clinical evaluation of a transtibial prosthesis. This evaluation is a tool to standardize the assessment process and to improve patient care. PMID- 11488056 TI - Amputation. Preoperative assessment and lower extremity surgical techniques. AB - Amputation surgery is an important part of the treatment for severe limb disorders. The decision-making process must be done thoughtfully, remembering that blood flow is not the only issue. Many factors enter into the decision to perform a partial foot amputation or to perform a more proximal level amputation. Adherence to good surgical principles, proven techniques, and gentle soft tissue handling can make the difference between a successful and durable amputation or continued complications and frustrations. PMID- 11488057 TI - Postsurgical orthotic devices. AB - Custom-made orthotic devices are beneficial as preoperative and postoperative modalities. The decision to institute conservative treatment such as casting and orthotic appliances depends largely on the pathologic condition the patient presents with, the overall medical and social status of the patient, and the desired results the physician wants to achieve. Orthoses, prostheses, and bracing have evolved from tedious and labor-intensive leather and metal components to technically advanced plastics, composites, and computer-generated efficient and patient-friendly modalities. With the incorporation of human biomechanics and engineering as well as the introduction of human performance laboratories and the shared knowledge from these disciplines, the field of orthoses and prostheses continues to expand for the benefit of those who must rely on man-made devices to augment daily living activities. PMID- 11488058 TI - Transtibial amputation. Prosthetic use and functional outcome. AB - Annually, roughly 80,000 to 100,000 amputations are performed with a total cost of greater than $50,000 per patient. Amputations in general and TTAs in particular are worthy of investigation to provide a cost-effective functional outcome that provides high patient satisfaction. The process of prosthetic selection, functional outcome, and avoidance of common complications begins with preoperative education of the patient and communication with the rehabilitation team. Determining an individual patient's perceptions of the process and his or her desired functional needs can help establish a baseline for prosthetic prescription and patient education. With the apparent difficulty in predicting who will be functional with a prosthesis, it seems prudent to initiate prosthetic training and fitting with all patients who have a moderate potential for use. Because many patients undergoing this operation have significant illness, as judged by the high mortality rate at follow-up, it is important to begin this process early to promote the highest recovery of functional independence. With the continual improvement in prosthetic components and postoperative management, it seems possible for the ill, elderly patient to regain near preoperative functional independence with the use of a prosthesis. PMID- 11488059 TI - Orthotic devices. Degenerative disorders of the foot and ankle. AB - Orthotic devices are an effective method of treating degenerative arthrosis of the foot and ankle. When combined with appropriate medications and activity modifications, operative intervention may be postponed or rendered unnecessary. A thorough comprehension of relevant anatomy, biomechanics, and available devices is crucial to maximize the benefits of this modality. PMID- 11488060 TI - Lower extremity orthoses and applications for rehabilitation populations. AB - There are many psychologic effects to trauma and disease. Patients often exhibit a grief reaction and loss of identity in the early stages of recovery. Fear and anxiety about the physical condition and future independence can contribute to feelings of separation and loneliness in patients. Loss of self-confidence and decreased self-esteem follow patients' efforts to overcome permanent decrements in functional abilities. Early intervention with an effective multidisciplinary treatment program can enable patients to perform mobility and self-care tasks. Functional independence and quality-of-life issues are important, and realistic individual goals must be identified and addressed throughout the rehabilitation process. The improvements in overall quality of life, provided by a multidisciplinary rehabilitation program, are as important as saving lives. Orthotic treatment programs are one component of an effective rehabilitation program and should be used to enhance the walking potential and abilities of patients with lower extremity dysfunctions. Ongoing home exercise programs and periodic reevaluation ensure the success of the program. Rehabilitation adds quality to the remaining life of patients by enabling them to return to or develop activities and hobbies as well as maintain higher levels of functional independence. The focus is always on the patient, and the best method to obtain success is an integrated team approach. PMID- 11488061 TI - Adaptive prosthetics for the lower extremity. AB - The potential for lifestyle recovery is tremendous for most lower extremity amputees. The amazing and ever-expanding array of adaptive prosthetics can help make the devastating loss of amputation more bearable for patients, their families, and their health care team. The new amputee, in a state of shock and grief, does not know what his or her prosthetic options are. It is crucial that the surgeon is knowledgeable about what the patient can have and what the patient needs to ask for. Dana Bowman stated: Ideally, the new amputee should say to their doctor, "I'd like my leg to be lightweight, flexible, durable, comfortable. I want to do sports or I want to ride bikes with my kids." Whatever it is they like to do. I was told I would never be able to wear two dynamic feet and that my sky diving days were over. I said, "Well how do you know? Can't I try?" It took years to find out what I could have and then to find people to help me get it. The prosthetic prescription the physician writes is the patient's gateway to the kind of prosthetics that will enable him or her to pursue the activities of their life. Often, new amputees end up with the bare minimum prosthesis, which can cause problems with comfort and mobility. A poorly designed or badly fitting prosthesis is as disabling as the actual amputation. When the surgeon can help the amputee and his or her family understand what kind of prosthetic choices are available, it establishes an optimistic outlook that is highly beneficial to the entire recovery process physically and mentally. "When I lost my leg, if someone would have told me that I could at least try to run again, that would have meant a lot," said Brian Frasure. "Getting that positive mental attitude is every bit as important as having good medical and prosthetic care." By asking probing questions about the patient's preamputation lifestyle and postamputation goals, the physician can write a prescription for truly adaptive prosthetics. The surgeon should consider the economic benefits of asking for a waterproof leg for an older adult that he or she can use for more stability in the shower and that can go a long way toward preventing fracture and other injuries. There are economic considerations for a younger amputee, asking for an energy-storing foot or an adjustable ankle so that he or she can be more active and independent and probably avoid the health risks associated with depression, inactivity, or obesity. If all patients received a contoured, flexible, dynamic socket from the beginning, years of medical problems and treatment for injury to the residual limb could be avoided. The initial prosthetic prescription is probably the best opportunity the patient ever will have to get the adaptive prosthetics that can meet his or her needs best. It also establishes a precedent with the payer for quality prosthetic care and begins educating all payers about what prosthetic patients can and should have. Insurance companies may resist the idea of a second prosthesis or of specialized components that increase the cost of the initial prosthesis. Physicians, patients, and the multidisciplinary team can work together, however, to pursue the most complete prosthetic coverage possible. For the prosthetic user, it is the difference between being restored to a full and active life or sitting back and becoming a spectator. PMID- 11488062 TI - CD56+/CD4+ lymphomas and leukemias are morphologically, immunophenotypically, cytogenetically, and clinically diverse. AB - CD56, a neural adhesion molecule, is a marker of natural killer (NK) lymphocytes as well as a subgroup of CD8+ T cells. Normal lymphocytes with a CD56/CD4 phenotype are scarce. Physiologic increases may occur in patients with immunosuppression, chronic inflammation, and autoimmune disorders. We report 4 cases of lymphomas/leukemias with the unusual CD56/CD4 phenotype. Two were of T cell and 2 of true NK-cell origin. The T-cell lymphomas had large granular lymphocyte morphologic features and splenomegaly. One patients had a benign course; the other died within months of the leukemia diagnosis. The 2 NK cell lymphomas had blastic morphologic features, initially involved skin, and had a very aggressive clinical course; 1 patient died of acute leukemia, and 1 had recurrence after bone marrow transplantation. Cytogenetic analyses did not show a consistent pattern of abnormalities. The NK lymphoma with acute leukemia had a t(2;5) but was CD30- and anaplastic lymphoma kinase negative. Although CD56+/CD4+ lymphomas/leukemias are a heterogeneous group, there may be a distinct subgroup of NK lymphoblastoid lymphomas of the skin, judging from our cases, as well as those previously reported. PMID- 11488063 TI - Bone marrow mast cell morphologic features and hematopoietic dyspoiesis in systemic mast cell disease. AB - Systemic mast cell disease (SMCD) cannot be distinguished from reactive mastocytosis (RM) by quantitation of mast cells in aspirate smears, and few studies have analyzed systematically the morphologic features of mast cells in SMCD vs RM. In addition, although SMCD is associated with myeloproliferative disorders/myelodysplastic syndromes (MPD/MDS), it is not known whether subtle signs of dysplasia or MPD can be found in SMCD, suggesting most cases are part of a dysplastic or myeloproliferative process. We compared 18 bone marrow specimens with SMCD with 10 bone marrow specimens from patients with RM. Mast cells in SMCD were more likely to show cytoplasmic hypogranularity, uneven granule distribution, and fusiform morphologic features. Eight cases of SMCD (44%) demonstrated MPD/MDS, and 9 cases (50%) showed subtle evidence of dyspoiesis, with megaloblastic change, nuclear budding of erythroid precursors, and/or atypical megakaryocytes. Mast cells in SMCD appear morphologically different from those in reactive proliferations. Dyspoietic features were present in most cases of SMCD, suggesting that SMCD is part of a spectrum of chronic myeloproliferative/myelodysplastic disorders. PMID- 11488064 TI - Clinicopathologic analysis of CD10+ and CD10- diffuse large B-cell lymphoma. Identification of a high-risk subset with coexpression of CD10 and bcl-2. AB - We analyzed 53 cases of diffuse large B-cell lymphoma (DLBCL) to determine whether expression of CD10 is a relevant biologic parameter. Tumor morphologic features were assessed semiquantitatively. Bcl-2 protein expression was studied by immunohistochemical analysis. The presence or absence of CD10 by flow cytometry was correlated with clinical and pathologic characteristics. CD10+ (23 cases) and CD10- (30 cases) DLBCLs were indistinguishable based on age, sex, extranodal presentation, B symptoms, clinical stage, morphologic features, or bcl 2 expression. However, cases with a CD10+ phenotype showed a significantly lower rate of complete remission. Cases expressing bcl-2 showed trends toward a lower rate of complete remission and poorer overall survival. Examination of CD10 and bcl-2 interaction revealed that the prognostic effects for both of these antigens were due to a subset of CD10+ bcl-2-positive cases. Compared with cases expressing one or neither of these markers, patients with dual-positive tumors had a poorer complete response rate to initial therapy and strikingly worse overall survival. While CD10+ and CD10- DLBCLs are similar with regard to a variety of clinical and pathologic features, CD10 and bcl-2 coexpressing tumors are an extremely high-risk subset based on response to therapy and overall survival. PMID- 11488065 TI - Immunophenotyping large B-cell lymphomas. Flow cytometric pitfalls and pathologic correlation. AB - Large cell lymphomas often challenge the diagnostic flow cytometrist. The purposes of this study were to improve our protocols for diagnosing large cell lymphomas and to correlate flow cytometric (FC) data with demographic and histologic features. We identified 63 cases of large B-cell lymphoma between January 1, 1995, and July 30, 1999, and reviewed the diagnostic slides and FC light scatter and staining patterns. The 51 lymphomas with adequate material for systemic review fell into 2 light scatter patterns: "clear cut," with large abnormal cells (high forward scatter relative to normal lymphocytes), 17 cases (33%); and "complex," 34 cases (67%). Clear-cut cases were more mitotically active (average of 42 vs 25 per 10 high-power fields), with higher cellularity. Apoptosis, geographic necrosis, and sclerosis were present histologically in many cases, regardless of FC findings. We conclude that morphologic features of large cell lymphomas do not predict which cases will be difficult to diagnose by FC. Gating strategies can be critical to improve the diagnostic yield. PMID- 11488066 TI - Transient myeloproliferative disorder and acute myeloid leukemia in Down syndrome. An immunophenotypic analysis. AB - Immunophenotypic analysis of transient myeloproliferative disorder (TMD) and acute myeloid leukemia (AML) using multiparameter flow cytometry might provide insight into their relationship. We retrospectively analyzed the expression of multiple lymphoid, myelomonocytic, and megakaryocytic antigens on blast proliferations in 18 patients with Down syndrome (DS; AML, 9; TMD, 9). The AMLs and TMDs shared several immunophenotypic characteristics. Blasts in all expressed CD45, CD38, and CD33; most AMLs and all TMDs were CD36+; and the majority expressed CD41 and CD61, suggesting megakaryocytic differentiation. The majority of cases were CD34+, CD14-, and CD64-. There was aberrant expression of the T cell-associated antigen CD7 in most AMLs and TMDs. CD56 was expressed aberrantly in 5 AMLs and 7 TMDs. The major difference between the disorders was the pattern of expression of myeloid markers CD11b and CD13; each was expressed in 8 AMLs but only 2 TMDs. Blasts were HLA-DR-positive in 3 AMLs vs 7 TMDs. Blasts in TMD and AML in DS have a characteristic immunophenotype distinct from AML in other settings. The immunophenotypic similarities suggest a biologic relationship between the disorders; however, distinct immunophenotypic differences also were observed. PMID- 11488067 TI - Pathology of autoimmune myelofibrosis. A report of three cases and a review of the literature. AB - We identified 3 patients with autoimmune myelofibrosis (AM) lacking American Rheumatism Association criteria for systemic lupus erythematosus (SLE). They had 1 or 2 cytopenias and lacked serologic evidence for SLE. Autoimmune features included psoriatic arthritis and positive direct Coombs test (DCT) result, DCT positive autoimmune hemolytic anemia, and synovitis with polyclonal hypergammaglobulinemia. Bone marrow biopsy specimens from each patient were evaluated by routine morphologic and immunohistochemical examination. They demonstrated marked hypercellularity (2 cases) or hypocellularity (1 case), moderate erythroid hyperplasia (all cases) with left-shifted maturation (2 cases), intrasinusoidal hematopoiesis (all cases), slightly to moderately increased megakaryocytes (2 cases), and grade 3 to 4 reticulin fibrosis (all cases). All lacked basophilia, eosinophilia, bizarre megakaryocytes, clusters of megakaryocytes, and osteosclerosis. Mild to moderate bone marrow lymphocytosis was noted in all cases. In 2 cases, increased small T cells and B cells formed nonparatrabecular, loose aggregates. AM is a clinicopathologic entity that may lack features of SLE. Loose aggregates of bone marrow T and B lymphocytes and the absence of morphologic and clinical features of myeloproliferative disease or low grade lymphoproliferative disease are clues that distinguish AM from better known causes of bone marrow fibrosis. PMID- 11488068 TI - Evaluation of the Prodesse Hexaplex multiplex PCR assay for direct detection of seven respiratory viruses in clinical specimens. AB - We evaluated the Hexaplex assay (Prodesse, Waukesha, WI) for the detection of 7 respiratory viruses (influenza A and B, parainfluenza 1-3, and respiratory syncytial virus [RSV] A and B). The Hexaplex assay was performed on 300 respiratory samples during the 1999-2000 respiratory virus season. Results of this assay were compared with shell vial cell culture and/or direct fluorescent antibody stain. The overall sensitivity and specificity of the assay were 96.6% and 94.1%, respectively. The respective sensitivity and specificity of the Hexaplex assay for detection of specific virus groups were as follows: influenza A, 98.6% and 97.8%; influenza B, 100% and 100%; and for parainfluenza viruses (1 3), 100% and 99.1%. The assay did not perform as well with patients infected with RSV: sensitivity and specificity were 91.0% and 98.6%, respectively. There are 2 major drawbacks to this assay: it is technically demanding (3-4 hours hands-on time), and it is expensive ($80-$90 direct cost). Nevertheless, because of the excellent sensitivity and specificity, the Hexaplex assay may be valuable in the diagnosis of respiratory viral infections in immunocompromised patients. PMID- 11488069 TI - Clinical and pathologic features of Mycobacterium fortuitum infections. An emerging pathogen in patients with AIDS. AB - The clinical and pathologic features of Mycobacterium fortuitum infection in 11 patients with AIDS were characterized. Nine patients had cervical lymphadenitis; 2 had disseminated infection. The infection occurred late in the course of AIDS, and the only laboratory abnormality seen in more than half of patients (7/11) was relative monocytosis. Absolute monocytosis also was seen in 4 of 11 patients. In both cytologic and histologic preparations, the inflammatory pattern was suppurative with necrosis or a mixed suppurative-granulomatous reaction. M fortuitum, a thin, branching bacillus, stained inconsistently in direct smear and histologic preparations. Staining was variable with Gram, auramine, Brown-Hopps, Gram-Weigert, Kinyoun, Ziehl-Neelsen, modified Kinyoun, and Fite stains. Organisms, when present, were always seen in areas of suppurative inflammation. Incorrect presumptive diagnosis, based on misinterpretation of clinical signs and symptoms or on erroneous identification of M fortuitum bacilli as Nocardia species, led to a delay in proper therapy for 7 of 11 patients. Definitive therapy after culture identification resulted in complete resolution of infection in all patients except 1. PMID- 11488070 TI - HER2 protein expression and gene amplification in androgen-independent prostate cancer. AB - The role of the HER2 receptor remains uncertain in the pathogenesis and progression of human prostate cancer. Previous studies have reported widely divergent rates for HER2 expression in primary prostate tumors, probably owing to significant methodologic differences in the studies. Few data exist about the frequency of HER2 protein overexpression and gene amplification in androgen independent prostate cancer (AIPC), although recent xenograft models suggest HER2 expression may be up-regulated in the transition from androgen-dependent to androgen-independent disease. We studied the role of HER2 protein in AIPC by immunohistochemical and fluorescence in situ hybridization (FISH) analyses on AIPC specimens using well-characterized and validated reagents. Fourteen (36%) of 39 specimens expressed HER2; however, only 2 (5%) had moderate (2+) expression, and 2 (5%) had high-level (3+) expression. Two (6%) of 36 specimens had gene amplification by FISH. These data suggest that HER2 protein overexpression and gene amplification are relatively uncommon in AIPC. PMID- 11488071 TI - The clinicopathologic significance of p53 and p21 expression in the surgical management of lingual squamous cell carcinoma. AB - The aim of the present study was to evaluate the clinicopathologic significance of p53 and p21 expression in lingual squamous cell carcinomas. Immunohistochemical staining was performed with p53 and p21 monoclonal antibodies on surgical specimens from 87 patients who underwent primary surgical treatment for lingual carcinoma between 1976 and 1996. We found positive expression of p53 in 45 (52%) of 87 cases and of p21 in 49 (56%) of 87 cases. There was no correlation of p53 and p21 expression with cancer stage, T stage, nodal metastasis, and tumor grade. Univariate analysis revealed that p21 expression, tumor stage, T stage, and nodal stage were significant prognostic factors for survival. However, only p21 expression and tumor stage were significant independent prognostic factors for survival in a multivariate Cox regression analysis. Overexpression of p21 but not p53 has prognostic value for survival in the surgical treatment of lingual carcinomas. The combination of stage with p21 expression is recommended for evaluation of prognosis and for management planning. PMID- 11488072 TI - WT1 is an integral component of an antibody panel to distinguish pancreaticobiliary and some ovarian epithelial neoplasms. AB - We investigated whether a panel of antibodies including WT1 could separate pancreaticobiliary and ovarian carcinomas by staining 64 pancreaticobiliary adenocarcinomas, 41 ovarian serous carcinomas, and 12 primary ovarian mucinous neoplasms with WT1, cytokeratin (CK) 17, CK20, carcinoembryonic antigen (CEA), and CA-125. Moderate or strong intensity reactivity in more than 25% of cells was a positive result. Of the ovarian serous carcinomas, 38 (93%) were WT1 reactive and 22 (54%) WT1 positive, 9 (22%) had CK20 reactivity, and 3 (7%) were CK20 positive in fewer than 50% of cells. All were CK17 or CEA nonreactive. Of the ovarian mucinous neoplasms, all were WT1 and CK17 nonreactive and 11 (92%) were CEA reactive, 8 (67%) CEA positive, 10 (83%) CK20 reactive, and 6 (50%) CK20 positive. Of the pancreaticobiliary adenocarcinomas, 19 (30%) were CK20 positive, 27 (42%) CK17 positive, and 52 (81%) CEA positive. All were WT1 nonreactive. A panel including WT1, CK17, CK20, and CEA is useful to distinguish pancreaticobiliary and ovarian serous carcinomas. Extensive CK17 reactivity is supportive of a pancreaticobiliary adenocarcinoma when the differential diagnosis includes ovarian mucinous neoplasm. None of the antibodies positively identified ovarian mucinous neoplasms. PMID- 11488073 TI - Immunohistochemical analysis still has a limited role in the diagnosis of malignant mesothelioma. A study of thirteen antibodies. AB - To identify the most accurate and useful panel to diagnose mesothelioma, we immunostained sections from 112 mesotheliomas, 18 adenocarcinomas, and 11 reactive pleural specimens with 13 antibodies. Positive results for mesotheliomas, adenocarcinomas, and reactive pleura, respectively, were CAM5.2, 111, 18, and 11; vimentin, 30, 3, and 3; HBME-1, 75, 10, and 8; thrombomodulin, 31, 2, and 2; calretinin, 43, 6, and 11; and CD44H, 68, 10, and 4. Positive results for adenocarcinoma markers in mesotheliomas and adenocarcinomas, respectively, were carcinoembryonic antigen, 1 and 15; LeuM1, 7 and 9; and Ber EP4, 5 and 12. All reactive pleura were negative. Positive results for markers to help distinguish mesothelioma from reactive pleura in mesotheliomas, adenocarcinomas, and reactive pleura, respectively, were epithelial membrane antigen, 76, 17, and 6; p53, 78, 16, and 9; P-170 glycoprotein, 37, 4, and 2; and platelet-derived growth factor receptor beta, 31, 1, and 2. The differential diagnosis of mesothelioma from adenocarcinoma is based on negative markers. Individual mesothelial markers are of low sensitivity and specificity for mesothelioma. However, diagnostic accuracy is improved by the use of antibody panels. To date there are no antibodies that help distinguish mesothelioma from reactive pleura. PMID- 11488074 TI - Histologic features of the liver in insulin resistance-associated iron overload. A study of 139 patients. AB - The aim of the present study was to describe histologic features of the liver in insulin resistance-associated hepatic iron overload (IR-HIO), defined as the association of metabolic disorders and hepatic iron overload. We included 139 patients in the study on the basis of one or more metabolic disorders and liver iron overload unrelated to usual causes. Liver biopsy specimens were reviewed, and histologic data were compared with those of a previously published, well defined population with genetic hemochromatosis. Iron overload was characterized by a mixed pattern with iron deposits in hepatocytes and sinusoidal cells. Steatosis was present in 59.7% of patients with inflammation in 32.4% of cases. Periportal fibrosis was found in 67.4% of patients. These patients were older, had higher sinusoidal iron scores, and had a higher prevalence of steatosis and inflammation than patients without fibrosis. Iron overload in IR-HIO was histologically different from that in genetic hemochromatosis. PMID- 11488075 TI - Cell biology and matrix biochemistry of chondromyxoid fibroma. AB - We studied matrix composition and gene expression pattern in chondromyxoid fibromas on the protein and the messenger RNA levels. We could clearly identify focal chondrocytic differentiation within chondromyxoid fibroma by the expression and deposition of type II collagen, which is a marker of chondrocytic cell differentiation. We also were able to show expression of collagen types I, III, and VI in the neoplasm. The major tumor portion was, however, characterized by the presence of hydrated proteoglycans and only minor amounts of collagens, a matrix composition responsible for the myxoid matrix appearance of most parts of these neoplasms. By analyzing cytoprotein expression, we found S-100 protein restricted to cells of the chondroid areas, suggesting S-100 protein staining to be of little help as a positive diagnostic marker for chondromyxoid fibroma. Our data show a specific matrix composition of chondromyxoid fibroma, not previously found in other mesenchymal neoplasms, including chondroblastoma, osteochondroma, enchondroma, and chondrosarcoma. This justifies chondromyxoid fibroma as a specific neoplastic entity, both clinically and biologically. PMID- 11488076 TI - Second opinion. PMID- 11488077 TI - Chronic external otitis. AB - Chronic external otitis is probably a disease of mixed etiology; infection and hypersensitivity both play an important role. As such, these two components must be recognized and respected. Steroids are the mainstay of medical management. Antibiotics should be used cautiously and probably on an intermittent basis. The disease appears to be exacerbated by any manipulation of the canal, including aggressive cleansing. Medical therapy will slow the development of fibrosis in some cases, but it does not always prevent it. Once stenosis has progressed to the point that conductive hearing loss develops, surgery will restore hearing and prevent restenosis in at least 80% of cases. PMID- 11488079 TI - Surgery for chronic ear disease. AB - When surgically treating chronic ear disease, the physician strives to accomplish several surgical objectives. Chief among them is to achieve a safe ear. This article discusses specific techniques that the surgeon can employ to reach these goals, whether he or she is performing tympanoplasty, atticotomy, intact canal wall mastoidectomy, or an open-cavity procedure. The author has found that the techniques discussed in this article are useful, and sometimes even essential, to achieving surgical objectives and maximizing outcomes. PMID- 11488078 TI - Post-tympanostomy tube otorrhea. AB - Post-tympanostomy tube otorrhea is a common problem that is treated by both primary care physicians and otolaryngologists. Physicians should take a logical approach to managing this condition in order to prevent the development of antimicrobial resistance and to minimize healthcare expenditures. Once the diagnosis has been made, first-line therapy with fluoroquinolone drops, with or without suctioning, is preferred. If the condition does not resolve after a few days, suctioning is recommended and oral antimicrobial therapy can be initiated, depending on the clinical situation. Parenteral therapy is sometimes necessary for those very few patients who do not improve with oral and topical antimicrobial therapy and aggressive local care. The use of prophylactic drug therapy is controversial. PMID- 11488080 TI - [Prevention of hemoglobinopathies in Brussels: a necessity?]. AB - Haemoglobinopathies are the most frequent genetic diseases in the world. The estimated frequency of carriers in the world is of 200 millions while there is about 300.000 births per year of major forms of haemoglobinopathies. The neonatal screening of haemoglobinopathies performed at the Hospital Erasme on around 80% of births in Brussels since 1994 has demonstrated that the frequency of carriers of an abnormal haemoglobin is around 1.5% while more than 1/2.000 newborns has a major haemoglobinopathy. A strategy must be adopted to manage the haemoglobinopathies in Brussels. PMID- 11488081 TI - [Therapeutic approach to chronic hepatitis B and C in the dawn of the third millenium]. AB - We try to illustrate the latest developments in epidemiology, pathogenesis and natural history of hepatitis B and C virus infection. Practical management of the patient with chronic B and C liver disease is presented. Universal hepatitis B vaccination should be encouraged in order to reduce to zero morbidity and mortality attributable to liver disease and its complications. Patients at risk for hepatitis B or C infection should be screened and notified about their evolutive risk and the therapeutic possibilities. PMID- 11488082 TI - [Chronic pain in geriatrics]. AB - Pain is frequent in communicative or no-communicative, ambulatory, institutionalized or hospitalized veterans. It is associated with severe comorbidity so much more than chronic pain could be neglected and expressed of atypical manner or masked by the absence of classical symptoms in particular in case of dementia or of sensory disorders. Pain detection by clinic examination or by pain assessment's methods and adequate approach by pharmacological and non pharmacological therapies are essential for correct pain management. On pharmacological plan, the strategy of the O.M.S. landings is applicable owing to a more particular attention to secondary effects and drugs interactions. AINS must be manipulated with prudence. There are no reasons to exclude opioides from the therapeutic arsenal but with a reduction of the starting doses, a regular adaptation and a very attentive survey. In drugs of landing 2, tramadol reveals itself as efficient and better tolerated as the codeine and dextropropoxyphene has to be to avoid. The obtaining of a satisfactory result depends on a regular assessment of the pain in a context of polydisciplinar approach (physicians, nurses, paramedicals, other care givers). PMID- 11488083 TI - [Polymyositis associated with HIV immunodeficiency: clinical case and literature review]. AB - A 33 year old female, suffering from HIV infection, presents with diffuse myalgia, generalized muscle weakness and painless dysphagia. An extensive work-up allows to diagnose an HIV-related polymyositis; while well-known, this entity is however rarely observed. Technetium-99m MDP skeletal scintigraphy shows multiple extra-osseous accumulations of the tracer: these observations, as well as the differential diagnoses of muscular involvement during HIV infection, are discussed. PMID- 11488084 TI - [A new approach in techniques to treat urinary incontinence: TVT (tension free vaginal tape)]. AB - The indications and the surgical technique of a newly developed operation for the correction of female urinary incontinence (the Tension free Vaginal Tape) are described. This technique is based on a new etiological concept of urinary incontinence and ensures the patient a reduced morbidity (local or regional anaesthesia) and hospital stay (theoretically adapted for a one day clinic). Mid term results (84% success rate after a median follow up of 3 years) are promising but must be confirmed at the long term. We relate our experience of 30 cases with a success rate of 93% but entached with a high rate of postoperatory urinary urgency. This leads us to emphasise a strict adherence to the indications and an absolute respect of the technique as originally described. PMID- 11488085 TI - [Aseptic osteonecrosis of alcoholic origin: a case report]. AB - Alcohol-induced aseptic osteonecrosis is not infrequent but multifocal osseous destruction is very rare. Alcoholic patients often present lipidic metabolism perturbation with fat embolism and are susceptible to develop diffuse intravascular coagulation in terminal microcirculation of femoral and humeral heads. The authors report one case of multifocal alcohol-induced osteonecrosis in a 74 year-woman. She presented with bilateral osteonecrosis of the humeral heads and total osteolysis of the neck and the head of the hips. Hepatic cirrhosis and chronic pancreatitis were additional risk factors inducing intra-osseous thrombosis. PMID- 11488086 TI - [Anti-TNF alpha in rheumatoid arthritis]. AB - Thanks to the better understanding of the physiopathological mechanisms in action in rheumatoid arthritis (RA), new therapeutic weapons have appeared, which have dramatically modified our approach of the disease. These so-called "biological" therapies antagonize the action of the cytokine at the top of the cascade which maintains the synovial inflammation, and leads to the joint destruction, i.e. the Tissue Necrosis Factor alpha (TNF alpha). Several controlled clinical studies have clearly demonstrated their short and middle term efficacy and safety profile, and they will soon become at the Belgian clinicians disposal. They have a rapid and dramatic effect on the signs and symptoms of RA and they slow down the radiologic progression. Some questions remain unresolved concerning their place in the general therapeutic strategy against RA, due to the uncertainties of their use in the long run, and to their cost. PMID- 11488087 TI - [Internal medicine and ophthalmology. The eye and drugs]. PMID- 11488088 TI - [Graves-Basedow ophthalmopathy]. PMID- 11488089 TI - [Type 2 diabetes: a refractory pathology?]. PMID- 11488090 TI - [Angiomas and vascular tumors: cultural aspects]. PMID- 11488091 TI - Adverse events following yellow fever vaccination. PMID- 11488092 TI - Global surveillance of emerging and re-emerging diseases. PMID- 11488093 TI - Epidemiology of inflammatory bowel disease in Spain. PMID- 11488094 TI - Decision-making process in long term acute cholecystitis. AB - OBJECTIVE: To demonstrate that the categorization of different therapeutic options is essential for the decision-making applied to clinical management. METHOD: This paper compares the usefulness of percutaneous echo-guided cholecystostomy as the first therapeutic option in acute long-term cholecystitis versus the exclusive surgical or clinical management. RESULT: We have analyzed the usefulness of different therapeutic options in a case of long term acute cholecystitis and we have obtained the following results: isolated intravenous antibiotic therapy (0.76), surgery (0.73) and echo-guided cholecystostomy (0.93). This last option has provided good clinical-radiological results. CONCLUSION: Decision trees are a good methodological option as support of surgical and non surgical therapeutic procedures. PMID- 11488095 TI - Normal values and influence of anthropometric and demographic factors on ambulatory cutaneous electrogastrography in healthy volunteers. AB - INTRODUCTION: Cutaneous Electrogastrography (EGGc) is a technique used for recording the Gastric Electrical Activity by means of electrodes placed on the abdominal skin. The aim of the present study was to determine normal values in healthy volunteers as well as the influence of factors such as age, gender, Body Mass Index (BMI) and echographic location of the gastric antrum. METHODS: An abdominal echography was initially performed in 35 of the 44 healthy volunteers in order to locate the gastric antrum and to place along its axis the skin electrodes, whereas in the other 9, the electrodes were placed according to anatomical references. Afterwards, the echography was also performed in those 9 volunteers in order to have all the 44 baseline echographic records. In every case, a record of ambulatory EGGc over 60 minutes was obtained, along with other record over the same period after eating a standard meal. The analysis of data was conducted through visual inspection and combined computer analysis. RESULTS: Eighty per cent of the study subjects showed a dominant frequency ranging from 2 to 4 cpm during both periods. After the meal, the parameters showed a characteristic variation. A significant difference was found when comparing subjects under 40 years of age versus subjects between 40 and 60 years of age in terms of preprandial dominant frequency instability coefficient (DFIC) (p = 0.002) and bradygastria (p = 0.03). Subjects with BMI < 25 kg/m2 showed preprandially a smaller dominant power instability coefficient (DPIC) (p = 0.035) and a greater Dominant Power (DP) (p = 0.045). In subjects without echographic control, DFIC (p = 0.001), bradygastria (p = 0.016) and tachygastria (p = 0.02) were more frequent, with a shorter period of normogastria (p = 0.001) during the postprandial period. CONCLUSIONS: Normogastria is the predominant rhythm in healthy people, although brief dysarrhythmias can be recorded that do not have any pathological meaning. Age and BMI, but not sex, seem to influence the Gastric Electrical Activity. The percentage of gastric dysarrhythmias decreases when the electrodes are placed along the longitudinal axis of the gastric antrum through echographic control. PMID- 11488096 TI - Influence of donor post-reperfusion changes on graft evolution after liver transplant. AB - INTRODUCTION: The increase in indications for liver transplantation has meant that waiting lists are growing ever longer. For this reason, broadening the donor pool is a priority for most groups. OBJECTIVE: The objective of this study was to analyze the predictive value of post-reperfusion biopsy in the evolution of graft function after liver transplantation. PATIENTS: One hundred and forty-eight liver biopsies, obtained after graft reperfusion, were analyzed. Eight pathological variables and thirty-seven clinical variables of the donors were recorded. Risk factors for presenting primary graft non-function or dysfunction were studied with logistic regression models. Factors associated to the long-term graft failure were studied using Cox analysis and actuarial survival curves. RESULTS: Microvesicular steatosis greater than 50% was the only risk factor associated to graft dysfunction in the multivariate logistic regression model. Microvesicular steatosis greater than 30%, severe hepatocyte necrosis and presence of abundant neutrophilic leukocytes were risk factors associated to graft failure in the univariate study. Only steatosis remained as an independent risk factor in the multivariate study. These grafts also presented poorer long-term survival. Abundant polymorphonuclear infiltrate was associated to a higher frequency of biliary complications. CONCLUSIONS: Microvesicular steatosis implies a better evolution than macrovesicular steatosis. Neutrophilic infiltrate and hepatocellular necrosis lead to poorer initial graft function and reduced long term survival. PMID- 11488097 TI - Is drug-induced hepatotoxicity unavoidable? PMID- 11488098 TI - [Complicated Morgagni's hernia treated as an emergency]. PMID- 11488099 TI - [Recurrent benign intrahepatic cholestasis]. PMID- 11488100 TI - [Unusual presentation of gastric adenocarcinoma]. PMID- 11488101 TI - [Small-cell desmoplastic intra-abdominal tumor]. PMID- 11488102 TI - [Vibrio mimicus gastroenteritis: report of 2 cases]. PMID- 11488103 TI - [Acalculous cholecystitis caused by Serratia marcescens]. PMID- 11488104 TI - Epidemiology of inflammatory bowel disease in Spain. A systematic review. AB - BACKGROUND AND OBJECTIVES: Incidence and prevalence of Inflammatory Bowel Disease (IBD) differ considerably depending on the geographical region studied. In Spain, both Ulcerative Colitis (UC) and Crohn's Disease (CD) seem to have experienced an important change in terms of incidence during the past decades. Our aim was to systematically review the experience in Spain regarding the epidemiology of IBD, in particular its incidence, its geographical differences and its evolution over time. METHODS: A bibliographical search till January 2000 was conducted in Internet using the PubMed search motor and the following descriptors or key words: "Crohn's disease", "Ulcerative colitis", "Inflammatory bowel disease", "Spain" and "Spanish". Only publications providing data on IBD incidence among Spanish patients were included. References from articles that fulfilled the selection criteria were also reviewed. The endpoints considered in the review were IBD incidence rate, its geographical differences, and its evolution over time. RESULTS: UC incidence reported by different studies performed in Spain ranges from 0.6 to 8 cases/10(5)/year, with a mean value of 3.8. CD incidence rates in Spain ranges from 0.4 to 5.5 cases/105/year, with a mean value of 1.9. Some studies have shown differences in IBD incidence between regions in our country. Many studies have reported an increase in IBD incidence in Spain over time, although such increase is more evident in the case of CD. CONCLUSIONS: UC incidence in Spain ranges from 0.6 to 8 cases/105/year, with a mean value of 3.8. CD figures range from 0.4 to 5.5 cases/105/year, with an average rate of 1.9. IBD incidence seems to be relatively different depending on the geographical region studied. Finally, several authors have demonstrated an increase in IBD incidence in Spain over time, although such increase seems to be more evident in the case of CD. Consequently, IBD in Spain is no longer considered a rare disease, but a relatively frequent one. PMID- 11488105 TI - Functional dyspepsia: are there any rational grounds for therapy selection? PMID- 11488106 TI - Criteria used by general practitioners in prescribing prokinetic or antisecretory drugs in patients with functional dyspepsia. AB - OBJECTIVE: To analyze the clinical factors considered by general practitioners for the prescription of prokinetic or antisecretory drugs in patients with functional dyspepsia (FD), and to assess therapeutic outcomes and factors predicting effectiveness. DESIGN: Multicentric, prospective and observational study. PATIENTS: 1,021 patients with FD were included. One hundred and thirty-two (132) were excluded from the analysis because they were taking ASA or NSAID. Patients were classified according to their predominant symptoms as reflux, ulcer, dysmotility or non-specific. At the physician discretion, treatment with alkali drugs was prescribed to 38 patients, prokinetic drugs to 574, antisecretory drugs to 123 and a combined therapy to 154. One month later, patient self-perception of symptomatic improvement was evaluated in patients treated with prokinetic drugs and antisecretory drugs. RESULTS: 85% of the patients reported symptomatic improvement after one month of treatment. Patients with non-specific FD had lower improvement rates regardless of the drug used (prokinetic or antisecretory) (77%) compared to all the other types (p = 0.03). Prescription of prokinetics was associated to female gender (OR: 0.43; 95% CI: 0.28-0.66) and early satiety (OR: 2.5; 95% CI: 1.6-4.1). A longer symptomatic evolution (OR 0.92: 95% CI: 0.88-0.97) was the only independent predictive factor of a poor response to prokinetic drugs. CONCLUSIONS: Among patients with FD attended by general practitioners, female gender and early satiety symptom were associated to the prescription of prokinetic drugs. Early symptomatic effectiveness rates for prokinetic or antisecretory drugs alike were high (85%). Patients with non-specific dyspepsia or long symptomatic evolution showed less favorable symptomatic response to prokinetic drugs. PMID- 11488107 TI - C282Y and H63D mutations of the haemochromatosis gene in patients with iron overload. AB - OBJECTIVE: To determine the relevance of C282Y and H63D mutations of HEF gene in patients with iron overload. PATIENTS AND METHODS: Patients with iron overload referred to our Liver Unit were included in the study. The association of mutations to different diagnosis and their impact on the severity of the hepatopathy were explored. Sensitivity, specificity and positive and negative predictive values of mutations for the diagnosis of haemochromatosis were determined. RESULTS: The study included 78 patients with iron overload. The control group included 21 patients of similar age and sex ratio without iron overload nor hepatopathy. Twenty three patients had haemochromatosis, 22 alcoholic liver disease and 33 other diseases unrelated to iron metabolism. Seventy three per cent of patients with haemochromatosis were homozygous for the C282Y mutation. All the C282Y homozygous subjects had also haemochromatosis. Fifty three per cent of patients with alcoholic hepatopathy had some kind of mutation. This has been also observed in 70% of patients with iron-unrelated diseases. Such percentage was significantly greater than in the control group (24% with H63D mutation). C282Y homozygosity in patients with iron overload had a sensitivity of 73.9%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 89.6%. CONCLUSIONS: In our population, as in all the Western countries, haemochromatosis is mainly associated to homozygous C282Y mutation. The high frequency of mutations in patients with iron overload and without haemochromatosis suggests the involvement of such mutations in iron overload. PMID- 11488108 TI - Association of MALTectomy (appendectomy and tonsillectomy) and inflammatory bowel disease: a familial case-control study. AB - There is some controversy regarding the prevalence of tonsillectomy and appendectomy among patients with Crohn's disease (CD) and a lower rate of appendectomy among patients with ulcerative colitis (UC). However, some environmental and familial factors that could alter those figures have not been studied. OBJECTIVE: To explore the prevalence of MALTectomy (appendectomy and tonsillectomy) among patients with IBD, stressing those factors that may be significantly associated to it. METHOD: Age-and-sex matched case-control study in patients with IBD, their relatives and the general population. Two hundred and eighty seven cases were IBD patients (153 UC, and 134 CD), the "family control" group included 203 siblings and the population-based control group included 570 individuals. Potential confounding factors, such as smoking, educational level, oral contraceptive use, place of birth and residence up to the age of 15 years, were ruled out. RESULTS: Appendectomy and UC: 7% of UC patients had undergone appendectomy versus 20% (OR: 0.23; 95% CI: 0.11-0.5; p < 0.0001) of controls. Appendectomy rates in families with at least one case of UC were 17/153 (6.3%) and 61/306 (20%) in the control group (p < 0.001). Appendectomy and CD: Twelve per cent of CD patients had undergone appendectomy six months before the onset of the disease versus 17% among the control population (OR: 0.43; 95% CI: 0.29-0.95; p < 0.01). The frequency of appendectomy in families with at least one case of CD was 22/221 (10%), which was significantly lower (p < 0.05) than among the control group 45/264 (17%). No differences were found between IBD patients and familial controls. Tonsillectomy and CD: Forty six per cent of CD patients had undergone tonsillectomy versus 39% of control patients (OR: 1.77; 95% CI: 0.92-2.05; p = ns). Tonsillectomy and UC: Twenty eight per cent of UC patients had undergone tonsillectomy versus 39% of the population control group (OR: 1.07; 95 CI: 0.57 1.25: p = ns). In fact, no differences were found regarding the prevalence of tonsillectomies within families with IBD cases as compared to population controls. CONCLUSIONS: Appendectomy is not only less frequent among CD and UC patients, but also among their relatives, thus suggesting the existence of environmental and genetic factors with opposed etiological roles in IBD and appendicitis. PMID- 11488109 TI - Truncal vagotomy and pyloroplasty for non-complicated duodenal ulcer: results 22 26 years after surgery. AB - OBJECTIVE: To assess the long-term results of truncal vagotomy and pyloroplasty in patients with non-complicated duodenal ulcer. PATIENTS AND METHODS: Between 1969 and 1973, 210 patients underwent surgery for non-complicated duodenal ulcer. Of these, 92 were followed up for 22-26 years. In 67 the assessment was conducted at the outpatient consultation and in 25, by phone. RESULTS: In 57% of patients the results were good or very good. Postpandrial fullness, heartburn and diarrhea were reported by 59, 52 and 42% of patients, respectively. Ulcerative recurrence affected 18% of patients. Although the relapsing rate was smaller in women as compared to men (7% versus 20%), physical condition and quality of life after surgery in the latter group were much worse. Thirty-five patients (38%) underwent reoperation. The distribution of patients according to the modified Visick classification after all the reoperations was as follows: I = 22 patients; II = 30 patients; III = 23 patients and IV = 17 patients. CONCLUSION: Long-term results after surgery for duodenal ulcer were not very encouraging, so we suggest for these patients a medical treatment and just in exceptional cases of refractory ulcers we would recommend a surgical procedure with minimal long-term complications. PMID- 11488110 TI - Microbiota and gastrointestinal system. PMID- 11488111 TI - [Brown bowel syndrome]. PMID- 11488112 TI - [Idiopathic fibrosing pancreatitis in pediatrics]. PMID- 11488113 TI - [Fasciola hepatica infestation: infrequent cause of cholestasis]. PMID- 11488114 TI - [Conservatively treated primary lymphoma of the rectum]. PMID- 11488115 TI - [Histologic findings in intestinal wall after ileocolic anastomosis with Valtrac]. PMID- 11488116 TI - "Good surgery is good economy" (FD Moore). PMID- 11488117 TI - Outpatient laparoscopic cholecystectomy. Four years of experience. AB - OBJECTIVE: To evaluate the feasibility of an ambulatory laparoscopic cholecystectomy program. DESIGN: Prospective analysis of ambulatory cases, overnight stay percentage, unexpected admission rate, postoperative complications and readmissions. PATIENTS: 271 consecutive patients undergoing elective laparoscopic cholecystectomy. RESULTS: Ambulatory laparoscopic cholecystectomy was completed in 193 out of 271 patients, reaching a 71.2 percent for the whole series. Ambulatory patients increased along with the experience of the team from a 22.9 percent in 1997 to 91.4 per cent in 2000. Sixty four patients (23.6%) had to be hospitalized overnight. Finally 14 patients had to be admitted (5.2%). The percentage of postoperative complications was 7.7%. CONCLUSIONS: Ambulatory laparoscopic cholecystectomy is a feasible and safe procedure with no increase in morbidity that can be performed in up to 90 per cent of patients. PMID- 11488118 TI - Characterization of the safety, effectiveness and use of oral sodium phosphate. AB - OBJECTIVE: The objective of this study was to assess the tolerability, safety and effectiveness of an oral sodium phosphate solution as a colon preparation method prior to colonoscopy, compared to a standard polyethylene glycol preparation. EXPERIMENTAL DESIGN: A cohort-based multicenter, prospective, observational clinical study was conducted. PATIENTS: Twenty-nine centers from all over Spain participated in this study, including a total of 638 patients who met all the inclusion criteria established. RESULTS: The oral sodium phosphate solution has an effectiveness and safety profile similar to the standard poliethylene glycol preparation; however, due to its dosage, it is better tolerated by patients, with a statistically significant difference between both preparations. CONCLUSIONS: Based on the effectiveness, safety and tolerability profile of oral sodium phosphate solution in patients, it can be considered the preparation of choice before colonoscopy, since it allows a fast, effective and safe bowel cleansing in patients without contraindications. PMID- 11488119 TI - Treatment of left colon neoplasic obstruction by placement of self-expandable stents. AB - INTRODUCTION AND OBJECTIVE: Neoplasic stenoses of the left colon are most frequently caused by primary colon carcinoma, infiltration from an external tumour and great adenomatous polyps. These patients often develop obstruction as their first symptom, leading to emergency surgical procedures in adverse circumstances and without an appropriate intestinal preparation that might prevent primary anastomosis. Therapeutic options for this event, such as Hartmann's resection, subtotal colectomy or anterograde colon lavage are not always possible. In these patients a colostomy is performed that requires future reoperation for reconstruction of the intestinal transit. Transtumoral self expandable stenting followed by elective surgery might be the best option in these cases, as well as an alternative to surgery in non-operable patients. PATIENTS AND METHODS: Twenty four patients treated with this procedure in the past four years were divided in two groups. In group 1 (14 patients), the stent was placed as a permanent and palliative measure for the management of the disease. In group 2 (10 patients), the stent was placed temporarily for the management of the intestinal obstruction and latter the patients underwent elective surgery with fully preoperative and extension study and an appropriate preparation of the colon in order to allow reliable primary anastomosis. RESULTS: There were no hospital mortality nor stent migrations. There was only one complication (perforation caused by the stent) that required emergency surgery, but with any further complications. Failure to place the stent occurred in one patient. CONCLUSIONS: Self-expandable stents relieve neoplasic colon obstructions and allow to complete the study protocol, followed by elective surgery associated to less morbi-mortality. In patients with advanced or irresectable cancer, they provide a palliative and safe alternative to surgery, with satisfactory results. PMID- 11488120 TI - Effectiveness of hemorrhoidal treatment by rubber band ligation and infrared photocoagulation. AB - OBJECTIVES: To demonstrate the effectiveness of the treatment of internal hemorrhoids with rubber band ligation (RBL) and infrared photocoagulation (IRC). PATIENTS AND METHODS: From march 1996 to december 1999, we prospectively studied 358 patients with a total of 817 hemorrhoid groups and a follow-up period of 36 months. Distribution according to gender and age was: 210 men with a mean age of 46 years and 148 women with a mean age 45.8 years. The mean number of hemorrhoids treated per patients was 2.3. All of them had complete a follow-up protocol at 15, 30, 60 and 180 days and at 12, 24 and 36 months. Rubber band ligation was performed with McGown ligator and suction pump, placing the band at the base of the hemorrhoid. For the infrared coagulation we used a Lumatec coagulation system, applying at least four shoots around each hemorrhoid, with an exposition time ranging between 1 and 1.5 seconds. Treatment was considered effective when patients became asymptomatic (relief of pain, bleeding or anal itching) and the obliteration of hemorrhoids after the treatment was confirmed by anal inspection and anoscopy. RESULTS: Two hundred ninety five of 358 patients were treated with RBL (82.4%), this treatment being effective in 98% of the patients after 180 days and very good after 36 months. There were 6/295 relapses at 36 months (2%). All minor and major complications were observed within the first 15 days of treatment: rectal tenesmus in 96/295 patients (32.5%), mild anal pain in 115/295 (38.9%), self-limited and mild bleeding after the detachment of the bands in 30/295 (10%), and febricula in one patient. Sixty three of 358 patients were treated with IRC (17.6%). In this group, relapses were observed in 6/63 patients (9.5%) at 36 months, all of them with grade III hemorrhoids that required additional treatment with RBL. All the complications (inherent to the technique) were observed within the first days: mild anal pain in 40/63 patients (63.4%) and mild bleeding in 1/63 (1.6%). The treatment with RBL or IRC depended on the number of hemorrhoids and the hemorrhoidal grade. No significant differences were found regarding the effectiveness between RBL and IRC for the treatment of grade I-II hemorrhoids, while RBL was more effective for grade III and IV hemorrhoids (p < 0.05). CONCLUSION: RBL and IRC should be considered as a good treatment for all grades of hemorrhoids, due to its effectiveness, its cost-benefit and its small short and long-term morbidity. PMID- 11488121 TI - Gastro-oesophageal reflux disease: new therapeutic perspectives. PMID- 11488122 TI - [Gammagraphy with red blood cells marked with technetium 99m: early visualization of the gallbladder]. PMID- 11488123 TI - [Transendoscopic ultrasonography with miniprobes]. PMID- 11488124 TI - [Myofibroblastic inflammatory tumor simulating recurrence of pancreatic vipoma]. PMID- 11488125 TI - [Relationship between age and clinical course of upper gastrointestinal hemorrhage]. PMID- 11488126 TI - [Constipation and hypertransaminasemia as presentation form of hypothyroidism]. PMID- 11488127 TI - Eruptive cheilitis: a new adverse effect in reactive HIV-positive patients subjected to high activity antiretroviral therapy (HAART). Presentation of six clinical cases. AB - A variety of exfoliative cheilitis has been observed in reactive HIV-1 patients subjected to high activity antiretroviral therapy (HAART). The lesions exhibit exfoliation, crater formation, fissuring, erosions and/or the formation of papules, vesicles and blisters associated to erythema and edema. The condition is not included in the 1993 EEC Clearinghouse classification (1) of oral lesions associated with HIV infection. In an earlier series of 1899 patients (2), we failed to observe this pathology and have only found one similar case described in the literature to date (3). OBJECTIVE: We present a series of 6 patients with HIV infection and morpho-histological alterations of the labial semimucosa, subjected to HAART. MATERIALS AND METHODS: The 6 patients were selected from among 20 HIV-positive individuals treated in our Infectious Diseases Unit with a combination of nucleoside and non-nucleoside reverse transcriptase inhibitors and protease inhibitors requiring stomatological care for painful lesions of the lips and oral cavity. The study was conducted over a 6-month period between May and October 1998. An analysis was made of the case histories, CD4-positive cell counts and viral load. The stomatological explorations were completed with biopsies, hematoxylin-eosin staining and immunohistochemical studies involving AE1 and AE3 monoclonal antibodies, vimentin, protein S-100, carcinoembryonic antigen (CEA), laminin, CD8, HLA-DR, BM-1 and CD31 markers. At the time of detection of the oral lesions, the patients had received different combinations of the following antiretroviral treatments: stavudine (D4T), zalcitabine (DDC), didanosine (DDI), zidovudine (AZT), lamivudine (3TC), nelfinavir (NFV), saquinavir (SQV), ritonavir (RTV), hydroxyurea (HU), indinavir (IDN) and efavirenz (EFV). RESULTS: There were four males and two females (age range 31-42 years). The CD4-positive and viral load ranges were 70-330 cells/mm3 and 200 500,200 copies, respectively. Stomatologic manifestations: The oral clinical manifestations consisted of desquamation, cracks, fissures, scabs, ulcerations, edema, erythema, multiple punctate xanthomas (1 case), dryness and labial semimucosal thickening. Microscopic study: Microscopically, the lesions consisted of ulcerations with adjacent hyperkeratosis and suprabasal vacuolization accompanied by a dense lymphocyte infiltrate within the chorion. Inmunohistochemistry: The immunohistochemical study in turn revealed surface epithelial alterations with AE1-AE3 monoclonal antibody labeling. Membrane glycoprotein activation (laminin) was detected, along with the presence of cytotoxic lymphocytes (CD8) and activated lymphocytes (BM-1). CD31 labeling in turn indicated endothelial activation. PMID- 11488128 TI - Maxillary undifferentiated carcinoma with rhabdoid features. AB - The rhabdoid malignant tumor was individualized by Haas et al. as an independent entity within the kidney malignant neoplasms in 1981. From its histopathological characteristics, different cases of rhabdoid extra-renal tumor were documented. This aspect has been also recognized in a wide heterogeneous group of tumors (carcinoma, melanoma, mesothelioma and mesenchymal tumors). The diagnosis of extrarenal rhabdoid tumor is based on the presence of a proliferation of epithelioid cells with large nuclei, prominent nucleoli, abundant eosinophilic cytoplasm, and conspicuous cytoplasmic intermediate filaments, which compress the nuclei. In this article we present the case of a 50-year-old male patient with a malignant maxillary tumor with characteristic rhabdoid features. Its differential diagnosis will be discussed. The literature will also be reviewed. PMID- 11488129 TI - Trigeminal neuralgia associated to basilar artery dolichoectasia. AB - A case of trigeminal neuralgia attributed to basilar artery dolichoectasia is described. Underlying vascular malformations and tumors of the cerebellopontine angle can be detected in patients with trigeminal neuralgia by means of noninvasive techniques such as computed tomography and magnetic resonance imaging; such diagnostic tools are therefore indicated in patients with trigeminal neuralgia to discard possible central nervous disorders. PMID- 11488130 TI - Extraoral etiology of halitosis. AB - Halitosis is a frequent complaint which is estimated to be found in around 50 to 60% of the general population and that carries serious personal and social repercussions. Although the majority of cases are due to oral problems, it is considered that 10-13% of halitosis cases are of extraoral etiology. In these cases the responsibility of the general dental practitioner, who is frequently the first person to examine and treat these patients, is to refer the patient for evaluation to an otorhinolaryngologist in order to rule out the presence of chronic tonsillitis or chronic sinusitis. If the otorhinolaryngologist does not detect alterations concerning his specialty, the digestive system should be explored in order to detect gastric pathology, obstructions or inflammatory gastrointestinal processes, the liver to rule out hepatic insufficiency or cirrhosis, the endocrine system to exclude diagnoses of diabetes or trimethylaminuria, the airways to rule out bronchiectasis or pulmonary abscesses, and the kidney to eliminate possible renal insufficiency. Finally, in the absence of any systemic organic pathology, the possibility of halitosis of psychiatric etiology, which requires the patient's psychological profile to be checked by the corresponding specialist, should be considered. PMID- 11488131 TI - Hereditary epidermolysis bullosa. Dental management of three cases. AB - Hereditary epidermolysis bullosa (EB) is a mucocutaneous disorder characterized by the appearance of blisters and vesicles in response to minimum friction. The digestive mucosa is one of the most frequently affected regions--including the oral mucosa. Three types of EB have been established according to the histological level of the lesion. Thus, simple EB involves intraepidermal bullae that leave no scars, while junctional EB exhibit blisters between the lamina lucida and lamina densa of the basal membrane. These lesions heal leaving atrophy and involve important hypoplastic lesions in the dental enamel. In turn, dystrophic EB presents synechiae-forming subepidermal blisters--the recessive form being the variant involving the greatest oral lesions (microstomia, ankyloglossia, milium cysts and rampant caries). Three cases of EB are presented and their clinical-dental management difficulties are described. The oral manifestations are described, along with the dental treatments provided and the evolution of the periodontal indices over a two-year period following the application of hygiene-preventive and therapeutic measures. PMID- 11488133 TI - Adult rhabdomyoma in the masticatory area. New case presentation and review of the literature. AB - Adult rhabdomyoma is a rare neoplasm of mesenchymal origin which represents approximately 2% of tumors with skeletal muscle differentiation. There are only about 100 cases reported in the literature. It is a benign tumor occurring most frequently in the head and neck region, which affects predominantly male population of a mean age ranging from 54 to 60. The purpose of this article is to present a new case of adult rhabdomyoma located in the masticatory area, arising as a slow-growing mass bulging in the left-side jugal mucosa and temporal region. The most common location for adult rhabdomyoma is the pharyngeal cavity. This type of tumor appears as a solitary mass, though occasionally may be multifocal. This tumor being of low occurrence, correct diagnosis can prevent aggressive surgery. PMID- 11488132 TI - Effects of vasoconstrictors in dentistry upon systolic and diastolic arterial pressure. AB - INTRODUCTION: Vasoconstrictors associated to local anesthetics habitually used in dental practice have been reported to influence arterial pressure, in the same way as other factors such as patient anxiety or stress. OBJECTIVES: The present study investigates the variations in systolic and diastolic arterial pressure in three groups of patients subjected to conventional dental treatment in the form of molar extraction, with the aim of determining whether such variations are attributable to the use of a vasoconstrictor or to other factors such as patient anxiety. MATERIALS AND METHODS: A total of 43 healthy males with normal blood pressure readings were grouped according to the type of anesthetic solution employed: Group I (2% lidocaine with epinephrine 1:80,000), Group 2 (3% mepivacaine with epinephrine 1:100,000) and Group 3 (3% mepivacaine with no associated vasoconstrictor). In all cases the Hamilton Anxiety Scale was applied, with blood pressure recordings before and after anesthetic injection, during dental extraction, and at the end of treatment. RESULTS: No statistically significant differences were observed in the blood pressure variations among the groups according to whether or not a vasoconstrictor was associated to the anesthetic solution in these healthy subjects. In increased systolic pressure was observed during extraction in all three study groups. CONCLUSION: Our findings suggest that the use of a vasoconstrictor in association to the local anesthetic solution does not substantially modify blood pressure. On the other hand, the observed increase in systolic pressure in all three treatment groups may be attributed to patient anxiety. PMID- 11488134 TI - Alveolar distraction osteogenesis using the "Lead System" distractor. PMID- 11488135 TI - Treatment of xerostomia. A review. AB - Saliva plays a fundamental role in the maintenance of oral health. Xerostomia is defined as a subjective sensation of oral dryness that usually implies a marked decrease in saliva secretion and may be due to a number of causes. Oral equilibrium is affected as a result, and buccal problems may develop. The present study reviews the different treatment modalities available for the management of xerostomia, destined to stimulate the production of saliva by chewing, administering gustatory substances, prescribing sialogogues such as pilocarpine, or using saliva substitutes or artificial saliva. The adoption of one treatment option or other in turn depends on the cause underlying xerostomia and on the functionality of the salivary glands. The review also evaluates preventive therapeutic measures and addresses the complications that may develop secondary to oral dryness. PMID- 11488136 TI - Ayurvedic interventions for diabetes mellitus: a systematic review. PMID- 11488137 TI - Surgical treatment of coexisting cataract and glaucoma. PMID- 11488138 TI - The toxins of Cyanobacteria. AB - Cyanobacteria, formerly called "blue-green algae", are simple, primitive photosynthetic microorganism wide occurrence in fresh, brackish and salt waters. Forty different genera of Cyanobacteria are known and many of them are producers of potent toxins responsible for a wide array of human illnesses, aquatic mammal and bird morbidity and mortality, and extensive fish kills. These cyanotoxins act as neurotoxins or hepatotoxins and are structurally and functionally diverse, and many are derived from unique biosynthetic pathways. All known cyanotoxins and their chemical and toxicological characteristics are presented in this article. PMID- 11488139 TI - The influence of anticholinergic drug selection on the effectiveness of oximes against soman-induced supralethal poisoning in mice. AB - 1. The influence of anticholinergic drugs (atropine, benactyzine, biperiden) on the efficacy of monopyridinium and bispyridinium oximes (HI-6, BI-6, obidoxime, pralidoxime, methoxime) on soman-induced supralethal poisoning was studied in mice. 2. While methoxime combined with benactyzine or biperiden seems to be more efficacious in the elimination of toxic effects of supralethal dose of soman than its combination with atropine, the efficacy of the other oximes studied against soman-induced toxic effects is not significantly influenced by the anticholinergic drug selection. 3. On the other hand, there are big differences in the effectiveness of oximes tested as to their ability to eliminate toxic effects of soman at supralethal doses. 4. The findings support the fact that the choice of acetylcholinesterase reactivator is more important than the anticholinergic drug selection for the effectiveness of antidotal mixture in the case of prophylactic administration of antidotes. PMID- 11488140 TI - The concept of contagion in Chinese medical thought: empirical knowledge versus cosmological order. AB - Since ancient times epidemics have been a central topic in Chinese medical thought. The explanations for their emergence, spread and transmission, however, have ranged widely. Whereas much of the populace believed in transmission by demons, elitist medical theory, since at least the second century, has emphasized cosmological and meteorological factors. This paper introduces the different approaches to epidemics in general, examining the etymological, historical and medical literature of early Imperial times. It then traces two lines of tradition in Chinese medical thought: one supporting contagionism, the other opposing it. The controversy that began as early as the fourth century, peaked in the twelfth century and ended only with the modern era, reveals a widening gap in the elitist medical theory--the dichotomy of empirical knowledge and theoretical framework. PMID- 11488141 TI - The complementary roles of observation and experiment: Theodosius Dobzhansky's Genetics of Natural Populations IX and XII. AB - Theodosius Dobzhansky has long been recognized by historians as a pioneer in the combining of the 'field natural history' and 'laboratory experimentalist' traditions in biology (Allen 1994). The following essay analyzes two papers in his well-known Genetics of Natural Populations series, GNP IX and GNP XII, which demonstrate how Dobzhansky combined field and laboratory work in the pursuit of an evolutionary question. The analysis reveals the multiple and complementary roles field observations and experiments played in his investigations. But it also identifies several interpretive problems associated with the use of intervention that limited the effectiveness of his approach. The essay argues that these problems reflect a fundamental tension between the amount of control Dobzhansky had over the circumstances of his experiments and the applicability of his results to natural populations. It concludes that this trade-off represents an important distinction between experiments in biology and most other sciences. PMID- 11488142 TI - [Carl Correns' experiments with Pisum, 1896-1899]. AB - The circumstances under which classical genetics became established at the turn of the nineteenth century have become an integral part of the standard narrative on the history of genetics. Yet, despite considerable scholarly efforts, it has remained a matter of debate how exactly the so-called 'rediscovery' of Mendel's laws came about around 1900. In this situation, unpublished research records can be invaluable tools to arrive at a more substantial and more satisfying picture of the order of historical events. This paper makes extended use of the research protocols covering Carl Correns' hybridisation experiments with Pisum sativum between 1896 and 1899. The resulting reconstruction sketches the portrait of a scientist following a particular research question--xenia--struggling with his experimental material, and slowly building up an epistemic regime in which questions and observations could acquire a relevance which did not strike Correns when he first took note of them. The microhistorical gaze through the magnifying glass of research notes reveals the kind of delays that appear to be constitutive for empirically-driven thinking in general. The research notes of Correns help not only to make this point, they also display some of the intricacies and material peculiarities which characterise the experimental process of hybridisation and the particular type of inferences it allows one to make. PMID- 11488143 TI - Of moths and men: Theo Lang and the persistence of Richard Goldschmidt's theory of homosexuality, 1916-1960. AB - Using an analogy between moths and men, in 1916, Richard Goldschmidt proposed that homosexuality was a case of genetic intersexuality. As he strove to create a unified theory of sex determination that would encompass animals ranging from moths to men, Goldschmidt's doubts grew concerning the association of homosexuality with intersexuality until, in 1931, he dropped homosexuality from his theory of intersexuality. Despite Goldschmidt's explicit rejection of his theory of homosexuality, Theo Lang, a researcher in the Genealogical-Demographic Department of the Institute for Psychiatric Research in Munich, revived it, maintained Goldschmidt's association with it, and argued on its behalf in publications from 1936 to 1960. Lang's appropriation of Goldschmidt's theory did not depend on his resolution of the difficulties Goldschmidt had found with his own theory. Lang and Goldschmidt, I argue, had fundamentally different scientific and social commitments that allowed one to reject this theory of homosexuality and the other to accept it. PMID- 11488144 TI - [Zoonites and organic units: its origins from a specific lecture of Alfred Moquin Tandon (1804-1863) and Antoine Duges (1797-1838)]. AB - To analyse the origins of the appearance of the zoonite concept as a unit of segmentary organisation in zoology requires positioning oneself in the context of the first half of the nineteenth century, more specifically in the years 1826 1839. If the origins of the problem go back to Goethe, on the one hand, and to botanists such as de Candolle on the other, the thesis of Moquin-Tandon on the splitting of plants (1826) is an essential step. The hypotheses of multiplication and diversification of organs arranged according to a symmetry, also presides over his research on the Hirudinees (1827). The zoonite is an elementary organic unit which can multiple itself and is structured symmetrically. But what characterises it is that its composition confers on it a certain autonomy. In 1832, Duges regarded the connection between zoonite and total organism from the angle of organic conformity. He deduced from his observations the laws of multiplicity of organisms, of disposition, of modification and of coalescence. This last law would become vital to Duges in 1838 for explaining the evolution of the composed being as well as embryonic development. But zoonitism in the 1830s did not constitute a scientific theory, even if it would ultimately serve as a reference, in particular for the animal physiology of Edwards. PMID- 11488145 TI - [Use and results of universal newborn hearing screening with ALGO portable device]. AB - BACKGROUND: Congenital hearing disorder, which has an occurrence of 1 in 1000 newborn babies, is a health problem that should be taken seriously. Once it has been discovered too late, it leads to lifelong language, intellectual, emotional and social problems. The aim of this project was the critical evaluation of a universal, ABR-based newborn hearing screening, looking at the specificity and the success of the method used as well as the organisational requirements and costs involved with this method. METHODS: From August 1999 until March 2000, 464 newborn babies, born at the University Hospital Eppendorf in Hamburg, were examined with the Algo portable. Another 31 children, all younger than 6 months, were also examined with the Algo portable, because of suspected hearing loss. When a child got a refer twice by the Algo portable, the child was examined again, using the conventional ERA-machine (Evoselect) to confirm the suspected hearing loss and to assess its grade. RESULTS: 98% of the newborns at our hospital were examined. Up to now, out of 13 conspicuous children, the majority of which belonged to the risk group, 6 have been identified as unambiguously bilateral and 2 as unilateral hard of hearing. After an introductory period of two months, the average costs per examination could be set at 38.65 DM and app. 3200 DM per bilateral hearing loss discovered. CONCLUSION: A Universal Newborn Hearing Screening is necessary and can be executed with calculable effort with concern to personnel, time and finance. PMID- 11488146 TI - [Detection of loudness recruitment in patients with retrocochlear lesions using the "Wurzburger Horfeld" loudness scaling]. AB - BACKGROUND: The pathogenesis of hearing loss caused by cerebellopontine angle tumors such as acoustic neuromas is unknown. The lack of loudness recruitment is thought to be one of the features of retrocochlear hearing impairment. In contrast to conventional suprathreshold tests, the categorial loudness scaling using the "Wurzburger Horfeld" is a valuable tool to describe the individual perception of sound. The aim of the present study was to analyze the loudness growth rate in patients with acoustic neuroma. PATIENTS AND METHOD: Pure tone and speech audiometry as well as auditory brainstem response and bilateral categorial loudness scaling were performed preoperatively in 54 patients with acoustic neuroma. Loudness scaling was done in free field switching off the contralateral ear by using an ear-plug. RESULTS: An abnormal rapid loudness growth function was found in 38 of the 54 patients (70.4%) at least at one frequency on the tumor side. The contralateral side was effected only in 57.4% of the patients. The incidence of a recruitment depended on the frequency with a maximum at 4 kHz. The slope of the loudness function showed a tendency to increase with increasing hearing loss. CONCLUSIONS: Loudness recruitment is not a rare phenomenon in patients with acoustic neuroma. The underlying cause (a preexisting hair cell damage, hair cell changes resulting from an obstruction of the cochlear blood supply or a disruption of the cochlear efferents) still remains unclear. PMID- 11488147 TI - [Vibrant Sound Bridge System. A new kind hearing prosthesis for patients with sensorineural hearing loss. 2. Audiological results]. AB - OBJECTIVE: Implantable hearing aids present a new treatment modality for patients suffering from sensorineural hearing loss. The functional gain obtained with the partially implantable Symphonix soundbridge system was evaluated in a clinical study. The audiological results achieved with n = 34 patients over a period of up to three years are presented in this second part of the publication. PATIENTS AND METHODS: 34 patients have received the Symphonix Vibrant soundbridge system since February 1997. The average age at implantation was 47.2 years (minimum: 18.9 years; maximum: 80.3 years). All patients have had several years of experience with hearing aids, which, however, provided insufficient functional gain or could not be fitted with a conventional hearing aid for medical reasons (such as auditory ear canal problems). All patients fulfilled the audiological selection criteria as they had bilateral moderate to severe sensorineural hearing loss. As a rule, the ear with poorer performance was implanted. All patients were fitted with the audio processor eight weeks after the implantation. The pure tone thresholds, the functional gain, the monosyllable and sentence understanding (Gottinger Sentence Test in quiet and noise) were preoperatively and postoperatively assessed. Standardized self-assessment questionnaires were used to evaluate the subjective benefit (PHAB) and the quality of hearing (HDSS) as compared to the preoperative situation. Further hearing tests were performed after four weeks, three, six, nine, twelve, eighteen, twenty-four and thirty-six months postoperatively. During the observation period of up to three years the audioprocessor was updated several times, most recently with the fully digital three-channel-system Vibrant D. The results obtained were documented. RESULTS: Postoperatively, the pure tone threshold with the soundbridge system switched off did not change significantly in the implanted ear. All patients had a functional gain that was either comparable to the gain achieved with hearing aids or better. In particular speech-related frequencies showed improved amplification. The free field speech recognition tests revealed higher scores in quiet and in noise. The patients commended the natural sound quality, the lack of feedback, the absence of occlusion and distortion, the improved speech understanding in noise and the favourable cosmetic appeal. Only two patients failed to achieve better results as compared to their performance with conventional hearing aids. No complications, such as a deterioration of hearing due to inner ear damage or a conductive hearing loss, were observed in the long-term. CONCLUSIONS: The Symphonix Vibrant Soundbridge is a new and promising treatment modality for patients suffering from moderate to severe sensorineural hearing loss. Further improvement of the good results can be expected with improved coupling of the transducer to the ossicular chain and further development of signal processing. PMID- 11488148 TI - [Mondini dysplasia without functional impairment in the framework of a CHARGE association]. AB - A male child with fully developed CHARGE association was referred to our hospital several times because of choanal atresia and suspected hearing loss in his first two years of life. As far as they are a main symptom malformations of the ear appear frequently in CHARGE association. This report describes that an obvious anatomic dysplasia of the inner ear can have normal function, which is shown by objective measurement. METHODS AND RESULTS: Computed tomography of the petrous bone showed bilateral Mondini malformation of the cochlea combined with bulky deformation of middle ear ossicles. By the age of one year brainstem evoked response audiometry (BERA) was performed. The result demonstrated a hearing loss of 60 dB on the right ear, but no measurable hearing loss on the left ear. CONCLUSION: This case report proves that a Mondini-type malformation of the cochlea does not need to be combined with hearing loss. PMID- 11488149 TI - [Radiosurgery of recurrent acoustic neurinoma or: the elegant solution of a surgical problem?]. AB - BACKGROUND: Stereotactic radiosurgery has proved to be an effective alternative to microsurgical resection in treatment of acoustic neuroma. Still, microsurgery is considered by many to be the therapy of choice. In case of recurrence microsurgical resection is much more difficult because of scarring and has a higher risk of complications. Therefore in cases of recurrence the role of radiosurgery needed to be evaluated. PATIENTS AND METHODS: From April 1992 to July 1997 135 patients suffering from acoustic neuroma were treated at the Neurosurgical Department of the University Medical School of Graz by means of the gamma-Knife. 12 patients had recurrence after a single or several microsurgical resections. The age distribution was between 38 and 71 years with a mean of 57 years. The diameter of the tumors varied between 10.5 and 31.2 mm. RESULTS: In all 12 cases the tumors could be inactivated biologically in a mean follow-up period of 58.8 months by means of stereotactic radiosurgery. Tumor shrinkage was achieved in 3 cases (25%), central necrotic areas were observed in 8 cases (67%). No additional cranial nerve palsies occurred. CONCLUSIONS: Stereotactic radiosurgery has proven to be a safe and effective treatment option instead of repeated microsurgery. Stereotactic radiosurgery should be considered as the therapy of choice in cases of recurrent acoustic neuromas. PMID- 11488150 TI - [Electron microscopic studies of the rabbit nasal mucosa after short-term application of naphzoline nitrate]. AB - BACKGROUND: Vasoconstricting nasal drops are applied frequently. Especially early ultrastructural alterations of the vessels were not examined up to now in animal models. Our goal was the systematic investigation of the submucosal vessels and the epithelium after topic application of naphazoline nitrate (Privin) in an animal model. METHODS: Three times daily over 6 days 3 ml of 0.1% naphazoline nitrate solution (Privin) were instilled into the right nasal cavity of 5 rabbits. At the 7th and 14th day an incisional biopsy of the lower nasal turbinate was carried out in ketamine/rompun anesthesia. RESULTS: In the naphazoline group the capillaries showed an edematous endothelium with narrowed lumina. Thrombosis of the arterioles were frequently observed. The more cubic epithelial cells had deciliated areas with microvilli. Venules, submucosal glands and the surrounding connective tissue was normally configurated. Analogous findings were also observed after 2 weeks. CONCLUSIONS: The disturbance of the microcirculation shows impressive endothelial alterations. These structural changes can result in nonreversible mucosal damages. A regeneration time of more than 1 week is assumable. Our results should be considered critically in order to prevent damages of the nasal mucosa. PMID- 11488151 TI - [Acute and fulminant fungal sinusitis in immunosuppressed patients]. AB - BACKGROUND: Acute and invasive fungal sinusitis represent rare diseases which can lead to life threatening complications. Immunosuppressed patients are affected primarily. The expansion of transplantation medicine and the progress in therapy of malignant diseases of the lymphatic system are associated with an increase of opportunistic systemic mycoses. Therefore the otorhinolaryngologist is confronted increasingly with these problems and questions for surgery, especially if the symptom of a periorbital inflammation occurs as a sign for a beginning orbital complication and radiological signs of involvement of the paranasal sinuses exist. PATIENTS: We report exemplary about two immunosuppressed patients with an invasive and fulminant fungal aspergillosis of the paranasal sinuses. In spite of systemic antifungal therapy and surgical intervention, intracranial and systemic complications caused a lethal course. CONCLUSIONS: In immunosuppressed patients with clinical and radiological signs for a sinusitis and a periorbital inflammation an invasive fungal sinusitis should be considered. Pathogenic aspergillus species are the most common identified in fungal sinusitis. The disease with often lethal outcome requires a careful and fast diagnostic and therapy as well as interdisciplinary co-operation. If and when limited or extensive surgery should be performed remains, because of the rarity and the lacking experience with this disease, still a controversially discussed issue and depends on several factors: the kind of disease, the immunity, the subtype of invasive fungal sinusitis and the degree of tissue invasion. PMID- 11488152 TI - [Historical outline on the nomenclature of neck lymph nodes as a basis of neck dissection classification]. AB - BACKGROUND: The neck dissection classification is based considerably on the organization of the lymph nodes of the neck. Terminology and anatomical allocation of nearly 300 cervicofacial lymph nodes repeatedly changed since the beginning of the 20th century. METHODS: Analysis of the literature on neck lymph node organization with reference to the development of the neck dissection classification. RESULTS: The first fundamental nomenclature of the neck lymph nodes is founded on the work of Rouviere (1932). Suarez (1963) described the functional neck dissection on the basis of the fascial compartmentalization of the neck. Lindberg (1972) left the predominantly anatomically correlated grouping of the cervical lymph nodes as described by Rouviere and divided the lymphatic system of the neck on basis of pathophysiological mechanisms. The attention regarding the location of occult metastases led to the description of the selective neck dissection. Since the fundamental work of Shah et al. (1981) there was a multiplicity of more or less slight changes of the neck node regions. These changes were again basis for new neck dissection terminologies. A new classification was introduced in the year 2000 as the revised version of the American Head and Neck Society. CONCLUSIONS: The revised version of the neck dissection classification can reduce former controversies, particularly regarding an optimized intraoperative allocation of the lymph nodes and a simplified terminology of the selective neck dissection. With the goal of a standardization of the neck dissection forms it remains to be seen if the proponents of the functional neck dissection after Suarez consider the extent of the neck dissection in patients with N0 neck in favor of the selective neck dissection. PMID- 11488153 TI - [Angiocentric T/NK cell lymphoma: a special clinical-pathological entity of lethal midline granuloma. A case report]. AB - BACKGROUND: The term "midline granuloma syndrome" (MGS) is a clinical description of a broad spectrum of diseases, which are characterised by aggressive and progressive destruction of mucosa and adjacent structures of the midface and upper aerodigestive tract. After exclusion of granulomatous infections, rare granulomatous diseases and epithelial neoplasias, the differential diagnosis includes the following entities: Wegener's granulomatosis (WG), malignant lymphoma and idiopathic midline destructive disease (IMDD). Today there are doubts about the existence of IMDD. After exclusion of WG nearly all remaining cases presenting as MGS are peripheral sinonasal angiocentric T- and/or NK-cell lymphomas, which show a close association to Epstein-Barr virus infection and now are recognised as a special clinicopathological entity. The natural history of these lymphomas is characterised through a rapidly progressive course with a poor prognosis. PATIENT: A case of a 35-year-old male patient with an angiocentric nasal T/NK-cell lymphoma, which involved the left lacrimal cyst, the left maxillar and ethmoid sinus as well as the soft and hard palates, is presented. First clinical signs and symptoms were similar to chronic-recurrent sinusitis. For almost two years the patient was treated with systemic corticoids for suspected limited Wegener's granulomatosis. The patient underwent sinus surgery for pansinusitis three times. After development of midline destructive disease the diagnosis of angiocentric lymphoma was established. RESULTS: Soon after the diagnosis a combination high-dose radiochemotherapy was performed. The patient died only 3 months later because of multiorgan failure. CONCLUSIONS: Because of its poor prognosis the angiocentric nasal NK/T-cell lymphoma should included early into the differential diagnosis of the midline granuloma syndrome. Correct biopsy technic and in situ hybridization of EBV can be important for an early diagnosis. Therapy should be aggressive and consists of high-dose radiotherapy, which is most important to reach local tumor control, and combination chemotherapy, the use of which is presently in discussion. PMID- 11488154 TI - [Dyspnea caused by leiomyomatous tumor of the oropharynx]. AB - BACKGROUND: Leiomyoma and Leiomyosarkoma are not often found in ENT-diseases. PATIENTS: We report on a 62-year old female patient presenting with a 3 x 4 cm tumor in the oropharynx that had developed within a period of about three weeks. The patient complained of increasing dysphagia and dyspnea especially when lying on the left side. To secure breathing the patient had to be primarily ventilated by placing a needle percutaneously through the conical ligament and was then intubated. For this procedure the patient was positioned on the right side. The tumor measuring 3 cm, was removed surgically. RESULTS: The tumor turned out to be a leiomyomatous neoplasia of undetermined malignant potential. Since an unfavourable behaviour of such neoplasms cannot be excluded, patients with tumors of this type should be followed closely. CONCLUSIONS: In this case the tumor leads to an acute dyspnoea and a special procedure of anesthesia was necessary. Therapeutic considerations and differential diagnosis of this very rare tumor of the oropharynx are discussed by reviewing the literature. PMID- 11488155 TI - [Evidence-based medicine in ENT]. AB - Using a clinical example we discuss the reasons why some of our colleagues associate unpleasant feelings with the term Evidence-Based Medicine (EBM). We demonstrate what EBM can contribute to solve day-to-day clinical problems and discuss the six formal steps for implementation of EBM into the day-to-day practise. 1. To transform the clinical problem into an answerable 4-part question. 2. To answer this question based on your internal evidence. 3. To search for external evidence to answer this question. 4. To critically appraise the found evidence with respect to it's validity, importance and applicability. 5. To integrate the appraised external evidence into your existing internal evidence in order to come to a new decision if the additional new external evidence in convincing. 6. To assess the benefit which was gained for the patient. PMID- 11488156 TI - [Surgery of glomus vagale tumors]. PMID- 11488157 TI - [Scalenus biopsy or Daniels operation]. PMID- 11488158 TI - [Basic principles and clinical aspects of implantation and the course of pregnancy with special reference to habitual abortion]. PMID- 11488160 TI - The molecular basis of embryo implantation in humans. AB - The implantation of the human embryo is a double paradox, immunological and biological. The immunological paradox is that it consists of a heterologous graft in which the uterine immune system (via the cytokines) and the embryo's antigenicity (HLAG) collaborate to make possible both implantation and the maintenance of the pregnancy. The biological paradox arises because several different mechanisms must be successively implemented for these two epithelia to fuse and then for one to allow invasion by the other (that is, the for the endometrium to be decidualized by the trophoblast): preparation of the endometrium throughout the menstrual cycle under the influence of estrogens and then progesterone, with the involvement of growth factors (EGF, TGF and IGF), neoangiogenesis (estradiol, FGF and VEGF), recognition by the trophoblastic cells of the various components of the decidua and of the extracellular matrix (integrins and cadherin) and the progressive invasion of the decidua, to the depth of the spiral arteries (by the trophoblastic secretion of metalloproteases). A defective or excessive trophoblastic invasion can result in complications of pregnancy: early spontaneous miscarriage, preeclampsia and growth retardation of vascular origin in the case of defects, placenta accreta or percreta in the case of excess. PMID- 11488159 TI - [The implantation receptive luteal phase of the endometrium. On the current status of molecular and cell biology research]. AB - The biological aim of the differentiation and maturation of endometrial tissue compartments during any menstrual cycle is the achievement of suitable conditions for blastocyst implantation and the establishment of pregnancy. Infertility and early embryonic loss are frequently caused by insufficient endometrial differentiation. Even any incomplete receptivity stage of the luteal phase endometrium will prevent attachment and implantation. We have studied the physiological changes throughout an endometrial cycle to elucidate causes of endometrial insufficiency leading to subfertility or infertility. Up to now, the histological changes described by Noyes et al. are understood as classical diagnostic approaches. However, evidence is accumulating that molecular deficits of endometrial differentiation are by no means detectable histologically, and consequently ask for the research on new diagnostic methods and parameters. There are histochemical localizations of specific protein molecules, adhesion molecules and cytokines, which permit by far more detailed and significant molecular analyses than any classical morphological means could yield. Moreover, there are convincing arguments to use further biochemical assessments on proteins of the uterine secretions as specific diagnostic parameters. The electrophoretical resolution presents typical protein patterns, which in turn can be interpreted as characteristic reflexions of the functional phases of the endometrial cycle. What is demonstrated as the so-called adequate luteal phase protein pattern clearly is the product of the receptive endometrium, reflecting the "implantation window". This is established already two days after ovulation and persists usually eight further days, if the endometrial cycle is undisturbed (15th to 24th day of the cycle). PMID- 11488161 TI - [Using ultrasound imaging in implantation]. AB - Sonography represents one of the best methods of clinical investigation which might give information about the preimplantation prognosis by evaluation of the following parameters:--morphological parameters (with bidimensional sonography): thickness and structure of the endometrium;--functional parameters (with doppler): evaluation of uterine vascularisation. The prognostic value of the morphological parameters appears to be controversial:--thickness of endometrium: a linear correlation between endometrial thickness and implantationsuccess may not exist. However no pregnancy occurs without a minimum threshold value of 6 to 8 mm endometrial thickness;--structure of the endometrium: concerning the role of the endometrial structure, controversial studies are available. According to some authors, endometrial hyperechogenicity is incompatible with a pregnancy. However hypoechogenic endometrium with 5 well delimited layers is an excellent prognostic factor. The coloured pulsed Doppler allows a functional assessment of the uterus and represents a technical progress in sonography investigations since the early nineties. The evaluation of uterine vascularisation can take place at different levels of the uterine vessels.--The pregnancy rate is inversely proportional to the uterine pulsatility index (PI). There is a consensus concerning a threshold value of PI. With values above 3, the pregnancy rate drops significantly.--The presence of a protodiastolic notch and the absence of end diastolic blood flow have bad prognosis. The presence or not of a sub-endometrialen vascular flow may be the most important prognostic parameter. No pregnancy may occur by absence of these flows. The sonography gives not only important morphological informations about endometrium but also allows a study of the uterine vascularisation by coloured doppler. This allows a real assessment of endometrial potentials for implantation. Optimal conditions of implantation could be:--endometrium > 7 mm,- hypoechogenic endometrium with 5 well delimited layers,--uterine PI < 3,- presence of sub-endometrial vascular flow. PMID- 11488162 TI - [Endocrinologic aspects of habitual abortion]. AB - Disorders of the luteoplacental progesterone shift in the first and second trimester lead to a insufficient progesterone secretion, thus constituting an important cause of early and late abortions. The endocrinological disorders caused by ovarian malfunction normally occur between the 5th and 10th weeks of pregnancy. We can distinguish early and late disorders of the corpus luteum graviditatis from a relative luteal deficiency owing to ovarian hyperstimulation. The delayed shift, however, is caused by trophoblast disorders and disturbances of placentation, thus leading to low blood levels of progesterone. Progesterone replacement therapy for the duration of insufficiency leads to significantly reduced rates of abortion. An increase in the rate of malformations is not to be expected. PMID- 11488163 TI - [Treatment of high risk pregnancy with gravibinon]. AB - A retrospective study was carried out in 47 patients 22 to 43 years of age, with a mean of 32.3 years, with a total of 51 high-risk pregnancies. The patients were treated with Gravibinon starting at the 5th week of pregnancy, and continuing up to the 20th. Indications for treatment were low progesterone or low estradiol levels, or both; these patients made up Group I. Group II patients showed a rapid drop in the progesterone and estradiol values after stimulation therapy despite ongoing pregnancy. The third group comprised women with PCO syndrome, who had previously anovulation or corpus luteum insufficiency. The pregnancy rate was 66.7%. In 9.8% a biochemical pregnancy was diagnosed through the hormone measurements. These biochemical pregnancies were not influenced by Gravibinon treatment. In 3.9% an empty cavity was found. In 11.8% abortion occurred prior to the 16th week of gestations despite Gravibinon treatment. In 7.8% late abortions occurred. In one case pregnancy was terminated owing to chromosomal anomalies. The pregnancy rate in all three groups was nearly equal, with group I at 20%, group II at 21.4%, and group III at 20%. In these high-risk groups it was found that treatment with an estrogen/progestin combination such as Gravibinon appears to be an effective treatment of an endangered pregnancy. An increased risk of specific anomalies was not observed. PMID- 11488164 TI - [Immunotherapy for prevention of abortion and for improving implantation in extracorporeal fertilization]. AB - In women with a history of recurrent in-vitro fertilization failure (at least three transfers of good quality embryos), there is an increased prevalence of organo-specific and not organo-specific autoantibodies, similar to women with a history of recurrent miscarriage. It is still unknown whether these autoantibodies are causally related to the failure of implantation or merely reflect an underlying systemic disorder. Several therapeutic intervention for the improvement of implantation after in-vitro fertilization and embryo transfer have been tried, including prednisolone (10 mg/d), heparin and aspirin (80-100 mg/d) alone or in combination with immunoglobulins and active immunotherapy using partner's leukocytes. At present, these interventions in combination with IVF should be regarded as experimental, as controlled, randomized and prospective trials in a large number of couples are still lacking. PMID- 11488166 TI - [Thoughts about the Geneve-Evian Congress]. PMID- 11488165 TI - [Improving treatment outcome by LeukoNorm Cytochemia in patients with multiple, failed IVF or ICSI treatment cycles]. AB - OBJECTIVE: The aim of these studies was to investigate whether the leucocytic ultrafiltrate LeukoNorm Cytochemia, which is approved for the treatment of immunologically-based recurrent spontaneous abortions (RSA), improves treatment results in patients with repetitive IVF or ICSI failures. MATERIAL AND METHODS: Included in this study were patients with 3 embryo transfers, with at least 8 morphologically good-looking embryos of the categories a and b, and no conception. Excluded were patients with accompanying endocrine disorders, uterine malformations, and those exhibiting the presence of an antiphospholipid syndrome and of further autoantibody syndromes (e.g. antinuclear antibodies [ANA]). The patients were treated in three prospective, randomized studies. The studies differed with respect to the frequency and timing of the administration of LeukoNorm Cytochemia. RESULTS: Pregnancy rates in each of the study protocols were generally higher than in the normal treatment group. Because of the small number of patients (study I: 10 patients), the differences were not always significant. The results of study III, which is still ongoing and in which LeukoNorm Cytochemia has been given on 5 consecutive days starting with the day of oocyte retrieval, are more significant. In the treatment group of study III the pregnancy rate was 55% as opposed to a rate of 21.2% in the non-treatment group. CONCLUSIONS: The administration of LeukoNorm Cytochemia can significantly improve treatment results in patients with repetitive IVF or ICSI failures. Currently the most favourable results are observed with a dosage of 1 unit/10 kg on 5 consecutive days, starting with the day of oocyte retrieval. These results imply that, in the IVF or ICSI programmes, there exists a group of patients with disturbances in the embryo-maternal dialogue, and therefore no conceptions. Furthermore, the results demonstrate that the administration of LeukoNorm Cytochemia can improve the implantation rate of transferred embryos in these patients. We conclude that growth factors and cytokines synthesized and secreted by leucocytes have an important influence on embryonic implantation and growth. PMID- 11488167 TI - [Tobacco and kidney]. PMID- 11488168 TI - [Sclerosing peritonitis]. AB - The case presented in this study illustrates the peritoneal changes observed in long-term peritoneal dialysis (PD) patients. This male patient was on peritoneal dialysis (CAPD) for seven months before and 86 months after renal transplantation. Two episodes of peritonitis occurred during that time. The patient developed symptoms (ascites, gastro-intestinal disturbances, deteriorating general condition, inflammatory syndrome) four months after starting hemodialysis, one month after ablation of the PD catheter. Other potential causes (infection, malignancy, hepatitis, etc.) of these symptoms were ruled out following an exhaustive etiological work-up. A final diagnosis of sclerosing peritonitis was made, and the patient was started on corticosteroid therapy. Both morphological and functional alterations of the peritoneal membrane associated with long term PD and the detection of such alterations in everyday practice are reviewed here, along with possible etiological factors and therapeutic measures discussed in the literature. A better understanding of the pathophysiological mecHanisms underlying these alterations would make it possible to develop preventive measures, such as more biocompatible dialysates. PMID- 11488169 TI - [Molecular aspects of phosphate homeostasis in mammals]. AB - Renal phosphate reabsorption, the major determinant of phosphate homeostasis, is primarily dependent on dietary phosphate content and multiple hormonal factors. Over the last few years, the identification of sodium-dependent phosphate transporters in kidney, intestine and bone, as well as new insights into the molecular mechanisms involved in several hereditary hypophosphatemias, allow to set up novel phosphate reabsorption regulatory pathways. This review describes molecular players involved in these mechanisms, summarizes phosphate transport data in kidney, intestine and bone, and describes recent findings concerning the three most common hereditary hypophosphatemias. PMID- 11488170 TI - [Treatment of end stage renal failure by dialysis in Rhone-Alpes: changes over the period 1993-1999]. AB - Over a seven years period, the prevalence of end stage renal failure patients requiring dialysis in a French area of 5.6 10(6) inhabitants was determined by yearly studies. The number increased from 1693 to 2296 (+ 35.6%), a 5% annual increase in the prevalence rate from 311 to 407. No tendency toward a plateau was evident. The main observed changes in the prevalent population were an increasing age (59.4 to 63), mainly for in-center patients, and male/female ratio, the rising of vascular or diabetic nephropathies (de 26.4 to 31.7%), and a decrease in the percentage of in-center dialysis patients (64 to 57%). PMID- 11488172 TI - [The First International Congress of Nephrology Geneve-Evian, 1-3 September 1960]. PMID- 11488171 TI - [Cortinarius poisoning. Analysis of cases in the literature]. AB - Cortinarius spp. poisoning is characterized by a delayed acute renal failure (ARF). The main features of this severe poisoning are still poorly known and often overlooked. The aim of this review of the literature was a better description of Cortinarius spp. poisoning. METHODS: The main medical databases were searched and analysed. RESULTS: 245 cases were collected and 90 cases could be analysed in details. Gastrointestinal disorders appeared a few days after the ingestion of the mushrooms (median: three days). The renal phase is delayed (median: 8.5 days). Hepatic failure and muscular lesions are highly questionable. Treatment is supportive in half of the cases, ARF progressed towards chronic renal failure, which progressed in 70% of the cases towards terminal renal failure. CONCLUSION: Cortinarius spp. poisoning is severe. Ingestion of Cortinarius species must be systematically suspected whenever tubulo-interstitial nephritis is diagnosed. PMID- 11488173 TI - [Letter to Nephrologie]. PMID- 11488174 TI - [Conflict of interest]. PMID- 11488175 TI - [Life circumstances and quality of life of lesbians and homosexuals]. PMID- 11488176 TI - [Intelligence and IQ]. PMID- 11488177 TI - [Prediction of IQ among children with birth weight under 1, 501 gms]. AB - INTRODUCTION: As a group, preterm infants are at considerable risk of cognitive difficulties. However, predicting cognitive sequelae has proved to be difficult. The current study reports on prediction of IQ at age eight years on the basis of the children's perinatal and developmental status, as well as parental socioeconomic status. MATERIALS AND METHODS: The sample consisted of 104 infants with birth weight < or = 1,501 g (53 girls), recruited consecutively in the neonatal intensive care unit and followed up to age eight years. Perinatal status, early cognitive development, and parental socioeconomic status served as predictors. Cognitive ability assessed on two commonly used intelligence tests served as outcome. The participants did not represent a total population, and no control group was used. RESULTS: Regression analyses revealed that birth weight, the Bayley cognitive index at 39 and 56 weeks, and parental socioeconomic status all made significant and independent contributions to outcome. Though the Bayley index made no significant contribution at age 29 weeks, it was the only variable at age two that was related to IQ at age eight. INTERPRETATION: Perinatal data are generally of limited value for the prediction of later IQ among preterm infants. However, when combined with information about parental socioeconomic status and the infants' developmental status up to age 56 weeks, birth weight made a unique and significant contribution as a predictor of later IQ. At age two years the cognitive status of prematurely born children was sufficiently consolidated to yield a valid prediction of outcome. PMID- 11488178 TI - [Physical activity and breast cancer]. AB - BACKGROUND: Physical activity may affect hormonal concentrations that may have bearing on the risk of breast cancer. The aim of this study was to investigate whether physical activity affects this risk. MATERIAL AND METHODS: All women aged 51-60 invited over an eight-week period in 1997 to a pilot breast cancer screening in Norway were also invited to participate in this study (n = 10,513). Information was collected by questionnaire about intensity and duration of physical activity from the age of ten up to the present, in five-year intervals. The activity was converted to scores of Metabolic Equivalent Task (MET). RESULTS: 74% og those invited attended the screening; 76% og those attending also took part in this study. A total of 2,300 of women gave adequate information about physical activity; only 12 of them had breast cancer. Those with cancer reported less physical activity compared to the controls. Physical activity corresponding to > or = 15 MET/week at age 10-24 reduced the risk of breast cancer at age 51-60 significantly (OR = 0.2; CI 0.0-0.9). INTERPRETATION: The small number of breast cancer cases limits the validity of the results, though the data indicate that physical activity of > or = 15 MET/week at age 10-24 may reduce the risk of breast cancer at age 51-60. PMID- 11488179 TI - [Palliative treatment in gynecologic cancer]. AB - BACKGROUND: Data on the need for palliative care related to disease groups are very limited. MATERIAL AND METHODS: A retrospective analysis was performed on the hospital records of 228 patients who died from gynaecologic cancer during the 1988-1997 period. RESULTS: 76% of deaths took place in hospital; 12% at home. Median terminal hospital stay was 13 days. Symptoms from intestinal obstruction were dominating in 30% of cases and most frequently seen in patients with ovarian cancer. Palliative intestinal surgery was performed in 38% of patients with ovarian cancer, median survival being 5 months (range 10 days-7 years), perioperative mortality (within 30 days) 13%. Cachexia dominated in 27% and was more often present when death occurred at home. Assistance from a gastroenterologic surgeon, urologist, haematologist or anaesthesiologist was needed in 32% of cases. One in two patients received parenteral pain relief in the terminal phase, for a median period of five days. Palliative radiotherapy was given 21% of the patients, most often in cervical and endometrial cancer, and chemotherapy or hormonal therapy was used in 36%. Minor interventions like laparocentesis, pleurocentesis, tumour resections, and various forms of urinary deviations were frequent. INTERPRETATION: Hospital deaths are more common among patients suffering from gynaecologic cancer than among cancer patients in general (55% in Norway), and the need for multimodal hospital service is large. We see substantial benefits in the Norwegian model in which a gynaecologist experienced in oncology has comprehensive responsibility for the treatment, supported by other specialists and the primary health service. PMID- 11488180 TI - [Immunomodulating treatment in advanced heart failure--effect of intravenous immunoglobulin]. AB - BACKGROUND: Congestive heart failure is characterised by enhanced immune activation. Immune-mediated mechanisms may play a pathogenic role, hence the growing interest in therapeutic regimens that could modulate the immune response in heart failure. MATERIAL AND METHODS: In the present report we discuss the pathogenic role of immunological and inflammatory mediators in the pathophysiology of heart failure and discuss different treatment modalities with focus on our recent study with intravenous immunoglobulin. In that study 40 patients with symptomatic chronic heart failure and left ventricular ejection fraction (LVEF) < 40% were randomised in a double-blind fashion to receive therapy with immunoglobulin or placebo for a total period of 26 weeks. RESULTS: We found that intravenous immunoglobulin, but not placebo, shifted the cytokine balance in an anti-inflammatory direction, and that such a shift was associated with improvement in LVEF by 5 EF units. Functional capacity and haemodynamic variables also improved. INTERPRETATION: Our study supports the hypothesis that immunological variables might be of significant importance in the pathogenesis of heart failure and it suggests a potential for immunomodulating therapy in addition to optimal conventional cardiovascular treatment regimens in such patients. These issues are further discussed in the present article. PMID- 11488181 TI - [Sphincteroplasty in traumatic anal incontinence]. AB - BACKGROUND: Anal incontinence in young women is often caused by sphincter rupture from obstetric injury or anal fistula surgery. MATERIAL AND METHODS: From 1992 to 1997, 17 women were operated for anal sphincter injury at Aker University Hospital, Oslo, Norway. Data were obtained retrospectively from patient files. RESULTS: In 13 patients, the anal incontinence was caused by obstetric injury and in four by anal fistula surgery. In all patients the external anal sphincter was repaired with overlap technique. In addition, plication of the puborectalis muscle was performed in 15 patients, perineoplasty in eight, vaginoplasty in one and operation for genital prolapse in one. Nine patients had a temporary loop sigmoidostomy. 12 of 16 patients for whom postoperative symptoms had been recorded, reported improved anal continence. Eight of these had complete anal continence, whereas four still experienced some incontinence for gas and loose stools. On clinical examination, 11 patients had normal resting sphincter pressure and voluntary sphincter contraction. Six had weak voluntary contraction; two of these had normal resting sphincter pressure. INTERPRETATION: Traumatic anal sphincter injuries in young patients can seriously impair anal function. Results of surgical treatment are satisfactory in most patients. PMID- 11488182 TI - [Pathological findings in the eye of children with facial nevus]. AB - BACKGROUND: Port-wine stains distributed to the trigeminal area may be associated with eye and/or central nervous system complications. Visual loss may be prevented with screening for eye pathology at an early age with adequate intervention. MATERIAL AND METHODS: 45 children with port-wine stains in the trigeminal area were examined by an ophthalmologist under general anaesthesia. RESULTS: Seven of 30 patients with one or both eyelids affected had a choroidal vascular anomaly. Four patients had glaucoma. These four patients had both eyelids affected. Two children with a vascular anomaly had only upper eyelid involvement. INTERPRETATION: Patients with facial port-wine stains affecting the eyelids should be screened for eye pathology at an early age. PMID- 11488183 TI - [Future pain treatment]. AB - BACKGROUND: The recent rapid progress in pain research is due in large part to advances in genetics and cell and molecular biology. We now know that chronic pain (hypersensitivity due to inflammation or nerve injury) and acute nociceptive pain are different and must be treated accordingly. We will continue to reveal sophisticated mechanisms underlying different kinds of pain that can be targeted to inhibit nociceptive transmission and produce analgesia. METHODS: This review is based upon literature collected through the authors' own reading and through PubMed searches. New hopes for future pain treatments are discussed. Further, the impact of genetic factors on pain sensitivity and pain modulation are discussed, and conceivable therapeutic approaches based on genetic techniques are mentioned. RESULT: At the level of the peripheral nerve, many novel targets have recently been identified: the tetrodotoxin-resistant sodium channel, the vanilloid receptor and different calcium channels are very interesting. In the spinal cord, different approaches can be used: to either block excitatory input or to increase inhibitory control or to do both at the same time. The mechanisms for hypersensitivity are being identified and offer multiple possible targets for novel analgesics. INTERPRETATION: Many interesting targets for analgetics has emerged during that last few years, lending great hope for new and better (i.e. with less side effects) analgesic drugs in the future. PMID- 11488184 TI - [Anti-ischemic and infarction-reducing effects of angiotensin-converting enzyme inhibitors]. AB - BACKGROUND: Angiotensin-converting enzyme is probably involved in the pathogenesis and progression of atherosclerosis, both through an increase in vascular angiotensin II and by an effect on the degradation of bradykinin into inactive fragments. Moreover, angiotensin II has a prothrombotic effect and prevails in increased concentration in the blood of animals subjected to experimentally induced myocardial infarction. MATERIAL AND METHODS: We have evaluated the relevant literature (including animal experiments and human studies) describing the infarct-reducing and anti-ischaemic effects of angiotensin-converting enzyme inhibitors. We also refer to studies in which these drugs have reduced the progression of atherosclerosis. RESULTS: Angiotensin converting enzyme inhibitors probably have favourable effects in various ways. There are indications that they counteract ischaemia, reduce heart failure and prevent reinfarction. Experimental observations in animals also indicate that angiotensin-converting enzyme inhibitors limit myocardial injury, presumably in part through the effects of bradykinin. This is supported by some few clinical studies, including a Norwegian epidemiological study. Angiotensin-converting enzyme inhibitors reduced the progression of intima-media thickness in the carotid arteries. INTERPRETATION: Angiotensin-converting enzyme inhibitors have favourable biochemical and haemodynamic properties that may explain their beneficial effects in patients with coronary heart failure. PMID- 11488185 TI - [Norwegian breast cancer mortality rates and validity of Swedish mammographic studies]. AB - BACKGROUND: The claim that screening for breast cancer with mammography reduces breast cancer mortality is mainly based on the results from the Swedish two county trial (WE study), where the effect was reported to be 30% for the age group 50-69 years. The two-county trial has recently been criticised for inadequate randomisation and for not following the study protocol. METHODS: We do some simple calculations to study whether the WE study is robust for an alternative statistical analysis. We use stage-specific breast cancer mortality in the Norwegian population as the baseline mortality rate in Sweden. Then we study the expected reduction in overall breast cancer mortality in the WE study while we vary the mortality rate in stage 1 and the stage distribution. RESULTS: We show that a 30% reduction in overall mortality rate is in conflict with observed decline in mortality in stage 1 and the expected stage migration. One either has to decrease mortality in stage 1, or increase the reduction of tumours with distant metastases, or both, to much higher levels than those reported in Sweden to get a 30% reduction in overall mortality of breast cancer. CONCLUSIONS: Our study adds further evidence to the proposal that the WE study is biased and not valid. PMID- 11488186 TI - [Vitamin E and coronary heart disease--why the lack of effect in large clinical trials?]. AB - BACKGROUND: Oxidative modification of low-density lipoprotein (LDL) is likely to play a role in the development of atherosclerosis and coronary heart disease. In vitro studies, animal experiments and several observational investigations suggest that vitamin E is an effective preventive agent. Unfortunately, large clinical studies have failed to confirm these findings. MATERIAL AND METHODS: On the basis of literature studies, I discuss the lack of effect. RESULTS: The hypothesis that LDL must be oxidised to be really harmful, could be wrong. If oxidised LDL is an important atherogenic factor, vitamin E (alpha-tocopherol) might not be a powerful enough antioxidant, especially in the later stages of atherosclerosis. Maybe one needs to use a mixture of different kinds of vitamin E and co-antioxidants that can regenerate vitamin E to achieve an effect that is powerful enough? The effect of vitamin E might have been masked by n-3 fatty acids in some studies. The most plausible hypothesis is that only people with extraordinarily high oxidative stress will benefit from vitamin E supplementation. INTERPRETATION: Further research is needed to understand how oxidised LDL and vitamin E affect the development of coronary heart disease. In the meantime it is important to point out that a healthy diet with balanced amounts of antioxidants and enough n-3 fatty acids has been proven effective. PMID- 11488187 TI - [Low-dosage acetylsalicylic acid as primary prevention against cardiovascular disease]. PMID- 11488188 TI - [Quality of life as psychological well-being]. PMID- 11488189 TI - [Treatment of acute myocardial infarction in Norway, thoughts on the development]. PMID- 11488190 TI - [Breast cancer and comorbidity]. PMID- 11488191 TI - [Information on preparations containing codeine]. PMID- 11488192 TI - [Suspected large fetus in the last trimester of pregnancy--a difficult problem]. PMID- 11488193 TI - [A casuistic report on Lamotrigine]. PMID- 11488194 TI - One step back and one, possibly two, steps forward. The different approaches to Parkinson's disease. AB - Parallel approaches to the development of medical treatment often experience advances in one technology sector and simultaneous setbacks in others. The potential role of medical devices and biomaterials in the treatment of neurological conditions such as Parkinson's Disease is discussed here together with the contrast between the role of devices in tackling the consequences of disease and that of drugs and cell therapies that aim to cure it. PMID- 11488195 TI - Semiconducting biomaterials: current status and future perspectives. AB - The ability to truly interface living cells with intelligent semiconducting biomaterials will launch new post-digital technology involving biologically interfaced medical devices. This article examines the potential of porous silicon for use in cell-interfaced electronic devices. PMID- 11488196 TI - The technological edge. AB - An advanced 3D digital imaging system has been developed under the MedLINK programme that offers improved detection of breast cancer. PMID- 11488198 TI - The ideal inhaler. PMID- 11488197 TI - US inspections of clinical investigation sites. AB - If the United States (US) Food and Drug Administration (FDA) receives the funding that it has requested, it plans to increase inspections of clinical investigation sites both in the US and abroad. This article discusses US clinical investigation requirements and guidance documents that should be reviewed when preparing for FDA clinical site inspections. PMID- 11488199 TI - Harness technology and reduce cost. PMID- 11488200 TI - The US market outlook for 2001. PMID- 11488201 TI - Laser welding of thermoplastic materials. AB - The capabilities of the three main types of laser are compared and a new technique is introduced, which laser welds plastics using an infrared absorber to create a joint that is almost invisible to the human eye. PMID- 11488202 TI - Biological medical devices: it's all in the name. PMID- 11488203 TI - [Peripheral thromboembolic complications in heart transplantation: prevalence and review of the literature]. AB - Peripheral thrombotic complications were retrospectively analyzed in a series of 164 consecutive patients undergoing orthotopic cardiac transplantation (CT) at the University Clinic of Navarra from 1984 to 1999. The overall survival during the first year after CT was 82%. All patients, besides the immunosuppressive regimen, received antiplatelet treatment postoperatively. The prevalence of thrombotic complications was 18% (29 events in 25 patients): 21 (13%) corresponded to arterial thrombosis, mainly cerebrovascular, whereas 8 (5%) were venous thrombosis. The time between the CT and the onset of thrombosis was 2 +/- 1.8 years. Mortality in patients with thrombosis did not differ significantly from the global mortality. The analysis in relation to the presence of cardiovascular risk factors only showed statistical significant differences for the age (older in the group with thrombosis, p = 0.02). In conclusion, there is a high prevalence of peripheral thrombotic complications in patients undergoing CT despite the antithrombotic regimen administered. The lack of correlation with the traditional cardiovascular risk factors suggest additional mechanisms for thrombosis in these patients. PMID- 11488204 TI - [Effect of PSA nadir and the time to nadir in the disease-free period in localized prostatic carcinoma treated with radical radiation]. AB - OBJECTIVE: To evaluate the prognostic significance of PSA nadir (nPSA) and the time to nadir in disease free of recurrence (DFR) in localized carcinoma of prostate treated with radical radiotherapy (RTR). MATERIAL AND METHODS: From October 1984 to December 1998, 86 patients have been treated with prostate carcinoma. It was considered of Low risk those patients with PSA < or = 10 ng/ml, Gleason = 6 or stage T1-T2. Moderate risk: those with one elevated of the three parameters. High risk: two or more parameters. The treatment was carried out in a lineal accelerator using photons of 15 MV, with standard technique and frationation, administering a median dose of 66 Gy (58-75 Gy). It was defined disease free of recurrence (DFR), the time to clinical PSA or biochemical failure. This one was defined as the time starting from the date of nadir PSA to the second consecutive increase of PSA value after three separate serial measurements separated for at least one month. RESULTS: The median of initial PSA value was of 16 ng/ml (1-270), initial clinical stage T1-T2 (70p), stages T3-T4 (14p), and unknown in 2p. The median of Gleason score was 6 (2-10). According to the group of risk they were classified as: low risk in 16 patients (19%), moderated risk in 22 patients (26%), high risk in 21 patients (24%), and unknown in 27 patients (31%). Median nPSA value was 0.8 ng/ml (limits: 0-139) and the median time elapsed between the initial PSA and nPSA has been of 11 months (limits: 0-72 months). The actuarial DFR projected to five years in those patients with nPSA = 1 ng/ml was of 67% vs. 47% in patient with nPSA figures > 1 ng/ml (p = 0.0018). The PFD in patients with time to nadir (t nadir) < 12 months it was of 20% vs. 80% in patients with t nadir > 12 months (p < 0.0001). Multivariate analysis demonstrated that time to nadir (H.R: 0.11 p = 0.001), group of risk (H.R: 28.72 p = 0.020), and grade of differentiation (HR: 28.72 p = 0.010), were determinant to DFR. CONCLUSIONS: nPSA is an important factor to determine the objective response to radiotherapy. nPSA and time to nadir are prognostic factors that influences significantly on the DFR. The indication of adjuvant treatment in those patients with unfavorable prognostic factors such us those who do not reach nadir PSA < or = 1 ng/ml and time to nadir < or = 12 months, deserves the realization of a prospective study. PMID- 11488206 TI - [Congenital synostoses of the tarsus. Concept, classification, diagnosis and therapeutic approach]. AB - The incidence of congenital tarsal coalition is about 1%. It is recognized as the main etiology of painful, rigid flatfoot in the pediatric population. Talonavicular (50%) and talocalcaneal (40%) coalition are the most common presentation. We must suspect a tarsal coalition in a child with mechanical pain and shoes deformity. Rigid and painful planovalgus deformity are found in physical exam which led to describe in the pass this entity as peroneal spastic flatfoot. Lateral and oblique (35 degrees-45 degrees) radiographs must be practice to observe the coalition. The presence of a beak in the head of the talus or a half moon condensation image as the result of the superposition of the talus over the calcaneus are commonly described. CT-scan is also useful to delineate the size of the coalition and its location. Resection of the bar is the surgical treatment of choice. Excision of the coalition and interposition of fat or a graft must be tried in young patients in order to preserve foot biomechanical properties and to avoid long term problems associated with arthrodesis. Triple arthrodesis or subtalar arthrodesis must be used in older patients with degenerative signs in radiographs or in those cases of multiple coalition or if resection has failed. PMID- 11488205 TI - [Significance of human mitochondrial genome in medicine]. AB - Mitochondrial function is necessary for energy production, but also plays important roles in oxidative stress and apoptosis. Part of the complexes responsible for mitochondrial metabolism are encoded in mitochondrial DNA (mtDNA). Knowledge of the structure and function of mtDNA affords a better understanding of (1) the physiopathology of mitochondrial disorders; (2) the pattern of inheritance of mitochondrial diseases; and (3) the strategies that can be employed in the molecular diagnosis of these disorders. In the near future important breakthroughs are expected regarding the understanding of the cross talk between nuclear and mitochondrial genomes, and its relevance in the biogenesis and maintenance of mitochondria. PMID- 11488207 TI - [Sexual behavior and reproduction]. PMID- 11488208 TI - [Celecoxib]. PMID- 11488209 TI - [Can we use clopidogrel in patients with coronary stent?]. PMID- 11488210 TI - [Therapeutic cloning and stem cells]. PMID- 11488211 TI - [Determination of the type and subtype of the hepatitis C virus in chronic viral hepatitis patients in Hungary]. AB - The huge variability of hepatitis virus C is well-established. The geographical differences in its nucleotide sequence have important clinical significance by causing variant pathogenicities and affecting sensitivity to therapy. The authors pioneered the determination of the viral type and subtype in patients suffering from chronic viral hepatitis in Hungary. In this present work they report the applied methods and the results. In their virus serological laboratory was introduced the test "HCV Serotyping 1-6 Assay", which is based on the analysis of antibodies, in 1996. By this method they examined the samples of 127 patients and they found type 1 in 75.5%, type 4 in 25%, mixed types (type 1 + 2, 3, 4, 5, 6) in 9% and non-classifiable, non-reacting antibodies in 13% of the cases. Since 1999 they have used the combination of direct reverse-PCR of the viral antigen and reverse hybridization to type-specific specimens. These results, from 211 patients show that 6% belong to 1a, 85.5% to 1b, 3% to 1a + 1b, 1% to 1b + 2, 0.5% to 3 and 4% to mixed subtypes. Genotype 1b was associated with higher viremia. On the basis of the above they can conclude that 90% of the Hungarian population are infected by the most resistant 1b subtype of hepatitis virus C. It could explain the fact that only 20% of their patients with interferon monotherapy have become permanently virus-free. In view of these results they recommend combined, higher dose, long-lasting treatment in therapeutic protocols. PMID- 11488212 TI - [Early and long-term results of treatment for infected deep sternotomy wounds]. AB - A retrospective study and follow-up was undertaken to determine the optimal treatment of poststernotomy wound infections. Between January 1990 and April 2000 mediastinitis developed in 62 patients following 7458 heart operations (0.83%). Median age of 50 males and 12 females was 59.1 +/- 9.1 years (27-77). Mediastinal infections occurred within 14.2 +/- 10.9 days (3-90) following cardiac surgery. Most common symptoms and clinical findings were purulent wound drainage or dehiscence (50%), sternal instability (46.7%) and fever (35.5%). Most commonly isolated pathogens were Staphylococcus aureus and coagulase-negative Staphylococci which alone or together with other Gram-positive bacteria caused nearly 70% of all infections. Following the exploration and radical debridement, the wounds in two patients were left to heal by open granulation, 26 patients were initially treated by closed mediastinal suction--or irrigation, and 43 patients were treated with muscle and/or omental flap transposition (34 primarily, 9 following the failure of other methods). Recurrence of infections occurred in 16 patients: 10 from the "closed treatment" group (38.5%) and 6 from the "soft tissue flap" group (14%). Nine patients died during the hospital stay. The mortality rate was 15.4% following the closed method (4/26) and 14.7% following the treatment with flap reconstructions (5/34). There were 6 late deaths, non-related to recurrent infection. Healed wounds were obtained in 96.2% of all patients. About one third of patients noted continuous or intermittent chest pain or discomfort in the closed and the soft tissue flap transposition group. 22.6% of patients claimed sternal instability and 9.7% shoulder weakness- each of them underwent muscle and/or omental flap closure. Abdominal hernias or bulges were present in 4 patients (12.9%) following the use of abdominal flap for wound reconstruction. These results suggest that early debridement and closed method would be successful when employed soon after cardiac surgery. When reexploration is delayed for any reason or obvious extensive involvement of bone or cartilage is present, the wound should be reopened, debrided and treated with muscle and/or omental flaps. Patients in whom closed method fails could probably treated with soft tissue flap transposition soon after the reoperation. Long-term results of closed mediastinal drainage or lavage and flap closure are favourable to open granulation technique regarding wound healing and elimination of infection. PMID- 11488213 TI - [New intraoperative diagnostic methods in parathyroid surgery: intraoperative measurement of parathyroid hormone]. AB - In thirty-seven patients undergoing parathyroidectomy from November 1999 to July 2000 with diagnosis of primary hyperparathyroidism the intraoperative intact parathyroid hormone level were studied. Preoperative samples of blood were taken from a peripheral vein before the beginning the operation. The second sample was taken 5 minutes after removal of parathyroid adenoma. The plasma intact parathyroid hormone concentration was measured by quick IRMA method. The preoperative high PTH levels reduced after excision of adenoma to the mean 18%. 33 patients had normal PTH levels after 5 minutes of removing. The total measuring time of quick PTH assay was about 30 minutes. The QPTH assay was a quantitative adjunct for the surgeon to ensure a successful parathyroidectomy. When the intraoperative QPTH level is not reduced to 50%, then the exploration should be continued and the probability of reoperation is decreased. They propose the application of the routine intraoperative measurement of QPTH level in all parathyroidectomy. PMID- 11488214 TI - [Eating disorders after the political changes in the formerly communist Eastern European countries]. AB - The authors summarize the Central and East European epidemiological data of eating disorders. These demonstrate that eating disorders are not exclusively characteristic to Western societies. In this respect the comparison of newer data to those which were performed before the political changes in 1989-1990 is especially valuable. Formerly in a Hungarian university sample the prevalence of bulimia was 1-1.3% among females, 0-0.8% among males, and this was higher than the prevalence of 0.6% in Austrian females, or 0% in German Democratic Republic. After the social changes similar data were found in several East European countries. This corroborates the culture-change idea of eating disorders. The transition to a Western market economy and the process of globalization is an experiment to evaluate the effect of sociocultural factors. The values and norms (e.g. thinness ideal) come closer to the Western culture, and mass media have a significant impact in this process. PMID- 11488215 TI - [Adnexal torsion in childhood]. AB - The authors report the case of a 10-year-old girl who was admitted with hypogastric complaints. Based on the examinations followed that, laparotomy was carried out, during which the torsion of the right-side adnexum was observed. During the case report special attention is put on the differential-diagnostic difficulties of adnexal torsion. The importance of the case is that we should think of this relatively rare disease even in childhood, because the early diagnosis and adequate therapy make possible to avoid the surgical amputation. PMID- 11488216 TI - [Molecular basis of antibody specificity]. PMID- 11488217 TI - [Analysis of results of surgical treatment of malleolar fractures]. AB - Late results of operatively treated malleolar fractures were analyzed. 101 patients were operated on, 58 (57.4%) were available for follow-up. There were 5 patients with A-type fractures according to Weber's classification, 28 with type B and 25 with type-C fractures. One year or more after the treatment the patients were evaluated on follow-up checks where ranges of ankle dorsiflexion, plantar extension, pronation and supination were recorded, circumference of the injured ankle was measured and the patients were interviewed about subjective complaints. Normal range of motion of the injured ankle (normal range of motion represented by the healthy ankle) was found only in patients with type-A fractures while in other two groups the patients had significantly decreased motility of the injured ankle (P < 0.05). The circumference of the injured ankle was increased in most patients with 25.9% patients ankles being 20 mm or more thicker than the uninjured side. Only seven patients (12.1%) were without subjective complaints while other patients experienced pain with load, pain with weather changes, swelling of the ankle etc. Malleolar fractures still represent a significant surgical problem and only an early operation, anatomic reduction and fixation as well as proper rehabilitation guarantee good results. PMID- 11488218 TI - [Use of international databases for improvement of comprehensive care of patients with head and neck cancer]. AB - The aim of this paper was to examine the usefulness of CancerNet in the improvement of knowledge of general practitioners on specific types of cancer and the procedures for patient management, thus indirectly producing an impact on quality improvement in comprehensive health care of patients with the head and neck cancer. Data on 58 patients with the head and neck cancer were collected from medical records in 28 general practices of the "Novi Zagreb" Health Center and from GPs' information. The GPs' knowledge on cancer of the larynx and hypopharynx was examined before and after they had studied the information provided by CancerNet. The mechanism of transferring information from the clinical and public health levels towards GPs was examined on GPs having in their care patients with the head and neck cancer. The following methodological characteristics were determined to have had an impact on patient care: within the structure of health care, a possible contribution of this procedure was identified in completing patient medical documentation as well as GPs' specific knowledge; within the health care process, a possible impact of this procedure on the quality of monitoring of a population health status, on more regular referrals to specialist consultations, as well as on pain and anxiety management, and a higher quality education on elimination of risk factors in patients' lifestyles. In average, the usefulness was scored by GPs with 4.0 for improving the communication with patients, and with 4.3 for expanding the GPs' knowledge. Three fifths of GPs involved in the study considered the contact range provided by CancerNet sufficient. All GPs assessed such information as useful for GPs in Croatia. PMID- 11488219 TI - [Neonatal alloimmune thrombocytopenic purpura caused by anti-HPA-1a alloantibodies. Case report]. AB - We report the case of serologically proven HPA-1a NATP. The child was born after uneventful 4th pregnancy. Immediately after birth generalized petechiae and signs of gastrointestinal bleeding were present. Isolated thrombocytopenia with the platelet number of 29 x 10(9)/L was observed. Serological investigation (PSIFT and MAIPA) showed high titre anti-HPA-1a antibody and low titre anti-HLA antibody in mother's sera. Mother's platelets were HPA-1a negative and she was HLA DR 52 positive. Father's platelets were HPA-1a positive. Cross-match between mother's sera and father's platelets was positive. 24 hours after the introduction of corticosteroid therapy platelet number increased to 73 x 10(9)/L and 48 hours later to 155 x 10(9)/L. The child was treated by corticosteroids because the NATP was severe and antigen negative platelets (mother or donor) or IVGG were not available. According to data from the literature the efficiency of corticosteroid therapy in NATP is questionable, but in this case it provided sufficient increase of platelet number with the stop of newborn bleeding. PMID- 11488220 TI - [Acute allergic posttransfusion reaction in a patient with immunoglobulin A deficiency]. AB - The incidence of selective IgA deficiency in general population is 1:400 to 1:750. When IgA immunodeficient person receives transfusion of blood or plasma, a severe allergic reaction can develop. This report describes a patient who developed a severe acute allergic reaction after receiving just a few milliliters of RBCs. Afterwards, investigation of the blood revealed that the patient had an IgA deficiency. This type of allergic reaction can be prevented by transfusion of IgA-deficient blood components or RBCs from a non-IgA-deficient donor which have been washed several times. PMID- 11488221 TI - [Bibliometric indicators in evaluation of research activity. 1. Publishing and evaluation of research]. AB - Publishing of scientific results is the key phase in scientific work. Reviewing process is a common way of evaluation of published scientific papers, but bibliometric indexes (number of papers, number of citations, impact factor etc.) are used as an adjunct in the assessment of scientific performance in scientific and academic promotion. The article discusses the presence of Croatian medical journals and Croatian medical authors in international indexing, i.e. data bases. PMID- 11488222 TI - [Experimental allergic encephalomyelitis as an animal model for multiple sclerosis]. AB - Experimental allergic encephalomyelitis is an animal model for demyelinating autoimmune disease of central nervous system, whose clinical and pathological characteristics resemble those in human disease multiple sclerosis. Multiple sclerosis is a chronic inflammatory disease followed by central nervous system demyelination, which is the result of an autoimmune process followed by central nervous system infiltration with autoreactive lymphocytes, activated macrophages and by local production of proinflammatory cytokines. Finally, the destruction of oligodendrocytes and myelin sheath occurs, caused by immunologic effector mechanisms. In this paper we reviewed fundamental and new facts about the most used models, induction, clinical and immunological characteristics of the experimental allergic encephalomyelitis in comparison with multiple sclerosis. Aethology and pathogenesis of multiple sclerosis are still unknown, but experiments on its animal models have improved our knowledge, not only about multiple sclerosis, but autoimmune diseases in general. PMID- 11488223 TI - [Snoring and obstructive sleep apnea]. AB - Snoring and obstructive sleep apnea are similar disorders in etiology, with differences in clinical presentation and treatment. 5-20% of young and 40-50% of older population are affected by either of these disorders. Etiology of snoring should be sought in the anatomic irregularities of commencing portions of the upper respiratory and gastrointestinal tracts, obesity, alcohol abuse, use of some medications as well as in the sleep architecture disturbance. Treatment success depends on correctly established diagnosis and precise localisation of anatomic irregularity implicated in snoring. A wide array of diagnostic tests exist some of which are mandatory while the others are used on a case-by-case basis. Treatment is either conservative or surgical. The success rate is 30-90% depending on the type and the management of disorder. An algorithm for diagnosis and treatment of snoring and obstructive sleep apnea is shown at the end of the paper. PMID- 11488224 TI - [Telemedicine: who--what--how]. AB - Telemedicine, or medicine at a distance, is based on electronic communications and digital technology, so their development has led to the rapid development of telemedicine as well. Today, telemedicine covers many areas of health care, such as signal transmission on patient health state, image transmission, vertical and horizontal consultations among remote experts, interventions at a distance, teleeducation, medical patient monitoring and alarm, etc. The greatest advantages of telemedicine are increased access to health care, reduced costs and quality improvement. Telemedicine in Croatia has been developing parallel to the world achievements thanks to the well developed infrastructure and enthusiasm of the concerned professionals. PMID- 11488225 TI - [Identification of epileptogenic networks from modeling and nonlinear analysis of SEEG signals]. AB - This work is focused on the study of the organization of the epileptogenic zone (E.Z.) in humans based on the analysis of stereo-electroencephalographic (SEEG) signals with signal processing methods, and more especially those dedicated to the estimation of signal interdependencies. In order to evaluate quantities provided by these methods and in order to relate them to the notion of functional coupling between cerebral structures, we developed a neurophysiologically relevant model able to generate EEG signals from organized networks of neural populations. We showed that the model can produce realistic multichannel epileptiform signals (when compared to real SEEG signals) under certain conditions (excitation/inhibition ratio within populations, uni/bi-directional coupling between populations). In this paper, the model framework is used to evaluate the performances of nonlinear regression analysis as a method to characterize couplings between cerebral structures from the SEEG signals they produce. Two quantities, a nonlinear correlation coefficient and a direction index, respectively related to coupling parameters in the model (degree/direction) are presented. These two quantities are measured on real SEEG signals recorded in patients suffering from temporal lobe epilepsy and candidate to surgical treatment. Results show that the characterization of functional couplings leads to the identification of networks referred to as 'epileptogenic networks', which might be responsible for the triggering of seizures. These results also corroborate our previous results on the classification of temporal lobe epilepsies, showing that there exist recurrent seizure patterns that can be classified on the basis of interactions between medial and lateral neocortical structures. PMID- 11488226 TI - Diagnostic value of quantitative EEG in Alzheimer's disease. AB - The aim of this study was to determine the performance of several spectral indices of the EEG (ratios between fast and slow EEG activities) as descriptors of the EEG changes occurring at the onset and during the evolution of Alzheimer's disease (AD). These indices were calculated from quantitative analysis of EEGs recorded in AD patients and from a matched non-demented group of control subjects. One advantage of such indices is to be independent of the absolute value of power spectral densities, which may vary from subject to subject, another being to take into account fast EEG activities. Conventional statistic tests and Receiver Operating Curves (ROC) analysis were performed upon these data to determine the accuracy of the power ratios to discriminate a) between controls and patients (i.e., to detect dementia) and b) between subgroups of patients defined according to the Global Deterioration Scale of Reisberg (GDS). The defined ratios provided a good classification of AD patients for all cerebral regions except the frontal areas, because of eye movement artefacts; the results confirm the increase in slow activities and the concomitant decrease in fast activities early in AD patients. Moreover, our results demonstrate that these indices are adapted tools to perform a good discrimination between demented and non-demented patients in routine clinical practice. We therefore propose the use of these EEG power ratios to discriminate between different stages of Alzheimer's disease, and to perform long-term monitoring of AD patients. PMID- 11488227 TI - Ultradian and circadian body temperature and activity rhythms in chronic MPTP treated monkeys. AB - The body temperature and locomotor activity rhythms of seven 1-Methyl, 4-phenyl, 1,2,3,6-tetrahydropyridine (MPTP)-treated cynomolgous monkeys were registered over a week on two separate occasions over an interval of 2 months. Motor disability was absent in two animals and present in five: it was mild in one, moderate in two and severe in two. Both temperature and motor activity were recorded every minute using a radio telemetry system. Analysis of circadian rhythms revealed less robustness of the 24-hour circadian components of body temperature and locomotor activity with increasing motor impairment, and a fragmentation of the body temperature rhythm into 8 hour-period components. Both total activity and daytime activity correlated inversely with the degree of motor impairment. On the contrary, the monkeys did not show differences in night time activity. The proportions of variance accounted for by the body temperature and locomotor activity of 24 h + 12 h + 8 h components were correlated. Also, the average levels at which the circadian rhythm varies between body temperature and locomotor activity were correlated. The results were almost identical in the two 1-week recording sessions. The present study confirms individual differences in the vulnerability to MPTP of the nigrostriatal system of monkeys, suggesting that if a cumulative dose does not provoke stable motor alterations, this cumulative dose will not produce circadian body temperature and locomotor activity rhythm alterations either. Similarly, if a dose is able to produce motor impairment, this dose will also be able to produce circadian rhythm alterations. PMID- 11488228 TI - [Opioid addiction: P300 assessment in treatment by methadone substitution]. AB - The aim of this study was to assess cognitive functions in two clinical conditions, namely during heroin detoxification and during substitution treatment by methadone. Two groups of chronic heroin user inpatients, meeting DSM-III-R criteria for concurrent opiate dependence, were tested using an auditory oddball paradigm of P300. The first group (four women and six men) were drug-free and the second (five women, ten men) received methadone treatment. Patients were also compared to a control group of non-dependent healthy subjects (five women, nine men). The patients were recorded 6-10 days after the beginning of either detoxification or methadone treatment. There were significant P300 alterations in the two patient groups, with amplitude decrease and latency increase, at a time when self-reported signs of withdrawal were absent or minimal. Paradoxically, the reaction time was accelerated in the two groups of patients, who also showed increased discrimination errors. These abnormalities were found with a lesser degree in the methadone-treated group than in detoxification patients. PMID- 11488229 TI - Is pruritus in chronic uremic patients related to peripheral somatic and autonomic neuropathy? Study by R-R interval variation test (RRIV) and by sympathetic skin response (SSR). AB - The problem of pruritus in uremic dialysed patients remains unsolved. The etiology of pruritus has not been precisely explained, and sometimes no efficient treatment is available. The aim of this study was to analyse the relationship between somatic neuropathy and pruritus as well as the relationship between pruritus and dysautonomia. Fifty-one patients with end-stage renal failure underwent basic neurological examination, nerve conduction velocity studies, and pruritus assessment by means of a questionnaire. Two tests were used to assess the autonomic nervous system, namely the R-R interval variation test in basal and profound breath conditions (RRIV) and the sympathetic skin response (SSR). Pruritus was found in 63% patients of the sample. Most of them had clinical symptoms and signs of peripheral sensorimotor neuropathy and dysautonomia. About 59% of uremic patients revealed abnormally reduced RRIV. About 45% of patients had abnormal (delayed or absent) SSR. The pruritus in uremic patients occurred significantly more frequently (P < 0.01) in patients with paresthesia. A nonsignificant but sizeable trend towards a relationship of pruritus with hypohidrosis and pathological SSR results was also observed. There was no relationship between the pruritus presence and RRIV results. According to our results the activity of the nervous system might play an important role in the mechanism of uremic pruritus, but paradoxically this latter appeared more tightly related to somatic neuropathy than to autonomic dysfunction. Our results also suggest that SSR may become a useful technique for the assessment of autonomic dysfunction in uremic patients. PMID- 11488230 TI - Another look at coral reef degradation. PMID- 11488231 TI - UK alien crayfish threat. PMID- 11488232 TI - Reprieve for white abalone. PMID- 11488233 TI - Comparing episodic, chronic, fishing, and non-fishing impacts on resource populations. PMID- 11488234 TI - Fate and effects of polydimethylsiloxane (PDMS) in marine environments. AB - Polydimethylsiloxanes (PDMS) defoamers are used to improve process efficiency under extreme conditions during gas-oil separation, when other chemicals fail to perform. They are also used to reduce the oil content of process waters discharged to the marine environment, thereby serving an important function in reducing oil pollution. As a consequence of these applications small quantities of PDMS may also be released into the environment. This paper reviews the fate of PDMS in the marine environment and the extensive effect studies that have been conducted. These demonstrate the absence of adverse effects on a wide range of marine species. PMID- 11488235 TI - Accumulation of polychlorinated dibenzo-p-dioxins and dibenzofurans in sediments of the Venice Lagoon and the industrial area of Porto Marghera. AB - Two sediment cores collected from the central Venice Lagoon and five from the canals of the nearby industrial area were studied in order to assess the temporal evolution of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) delivery and present trends. The most polluted site is C12, in the Brentella Canal, with a total equivalent (TEQ) concentration of 2858 and 64,130 ng kg-1 at the surface and at 18-20 cm depth, respectively. High values can be also found in the Nord Industrial Canal and in the Salso Canal: 3000 and 2500 ng kg-1, respectively. Lagoon samples are much less contaminated, with TEQ downcore maxima of 22 and 53 ng kg-1, being influenced by pollutants stored in sediments of the canals only occasionally. Sediment chronology, based on 210Pb and 137Cs activity depth profiles, shows that the highest fluxes occurred in the 1970s and early 1980s, and that pollution is now decreasing. Homologue profiles can distinguish PCDD/Fs from different sources: combustion for octachlorinated dibenzo-p-dioxin (OCDD) and stripping of VCM for octachlorinated dibenzofuran (OCDF) alone, whereas several activities carried out in the past within the industrial area can be responsible of the overall contamination by furans. PMID- 11488236 TI - Temporal variation in the diversity and cover of sessile species in rocky intertidal communities affected by copper mine tailings in northern Chile. AB - Several coastal rocky shores in the northern Chile have been affected by the discharges of copper mine tailings. In spite of this, the temporal and spatial variation on the diversity and composition of their intertidal benthic communities has scarcely been studied. The objectives of the present study were to analyse and to compare quantitatively the temporal variation on the diversity, cover and composition of sessile species in rocky intertidal benthic communities of the northern Chilean coast, in relation to the presence of copper mine tailings. The results show that the drastic reduction on the sessile species diversity and the monopolization of the substrate exerted by the green algae Enteromorpha compressa, are common and permanent features of the intertidal rocky shores affected by copper mine tailings. Such spatial (between sites) and temporal (seasonal) variation of these changes has been associated with the relative concentrations of trace metals and inorganic particles of the mining wastes. Our results suggest that the mechanical effects of resuspended and settling tailings are a more likely cause. PMID- 11488237 TI - Magnetic properties and heavy-metal contents of contaminated seabed sediments of Penny's Bay, Hong Kong. AB - Magnetic properties and heavy-metal concentrations of vibrocore samples were found to be potential indicators of shipping contamination in seabed sediments in Hong Kong Harbour. Geochemical results of 74 vibrocores located off Penny's Bay on Lantau Island revealed an enrichment of heavy metals in the upper 1-2 m of the cores within the eastern part of the study site. Whole-core magnetic susceptibility measurements also showed a greater concentration of magnetic particles in the surficial layer of these cores. A significant correlation exists between the magnetic susceptibility and the concentrations of Pb, Zn and Cu, as well as the Tomlinson pollution load index (PLI). The proximity of these cores to a major navigation fairway and an anchorage site suggests probable contamination of the surficial seabed sediments by shipping-related wastes. A study of the magnetic properties of one of the cores, VT60, revealed a difference in the magnetic properties between the contaminated and the uncontaminated sediments. Samples from the contaminated zone exhibited relatively stronger magnetic remanence and susceptibility. The two groups of samples also formed distinct trends on the hysteresis ratios plot. A level of unusually high magnetic susceptibility within the contaminated zone was attributable to the presence of strongly magnetized granules, which were probably refuse from shipping-related activities. Geochemical and magnetic results were also conducted on samples of different size-fractions from this core. The < 63 microns fraction was found to contain a relatively higher magnetic susceptibility and greater heavy-metal content. PMID- 11488238 TI - A pre- and post-MARPOL Annex V summary of Hawaiian monk seal entanglements and marine debris accumulation in the northwestern Hawaiian Islands, 1982-1998. AB - Entanglements of Hawaiian monk seals, Monachus schauinslandi, were documented in the northwestern Hawaiian Islands (NWHI) from 1982 to 1998, and debris which presented a threat of entanglement was inventoried and removed from 1987 to 1996. A total of 173 entanglements was documented. The number of entanglements did not change after implementation of MARPOL Annex V in 1989. Pups and juvenile seals were more likely to become entangled than older seals, and became entangled primarily in nets, whereas entanglement of subadults and adults was more likely to involve line. The subpopulation of seals at Lisianski Island experienced the most entanglements, although Lisianski did not accumulate the most debris. Localized high entanglement rates may gravely affect individual monk seal subpopulations. Accumulation of debris has not diminished since implementation of Annex V, nor has occurrence of derelict drift nets abated since a 1989 moratorium. Debris washing ashore has likely been circulating in the North Pacific Ocean for some time. PMID- 11488239 TI - An oil spill-food chain interaction model for coastal waters. AB - An oil spill-food chain interaction model, composed of a multiphase oil spill model (MOSM) and a food chain model, has been developed to assess the probable impacts of oil spills on several key marine organisms (phytoplankton, zooplankton, small fish, large fish and benthic invertebrates). The MOSM predicts oil slick thickness on the water surface; dissolved, emulsified and particulate oil concentrations in the water column; and dissolved and particulate oil concentrations in bed sediments. This model is used to predict the fate of oil spills and transport with respect to specific organic compounds, while the food chain model addresses the uptake of toxicant by marine organisms. The oil spill food chain interaction model can be used to assess the environmental impacts of oil spills in marine ecosystems. The model is applied to the recent Evoikos Orapin Global oil spill that occurred in the Singapore Strait. PMID- 11488240 TI - Metal dynamics in an Antarctic food chain. AB - The concentrations of copper, zinc, cadmium, selenium and mercury were determined in eggs, muscle, liver, kidney and stomach content of nestilings and adults of the Antarctic petrel, Thalassoica antarctica, and its predator, the south polar skua, Chataracta maccormicki, from Svarthamaren, Dronning Maud Land, Antarctica. The dominant food of the petrels is krill, Euphausia superba. The results show relatively high levels of cadmium in krill, which is assumed to be the main reason for the high levels of cadmium in petrels and skuas. Cadmium is almost absent in eggs, but accumulates very rapidly with age in nestlings. The copper concentrations in livers of nestling petrels reach very high levels during growth. This may be seen in connection with physiological development processes. Mercury seems to be accumulated with age and between trophic levels. Among the nestlings, the mercury levels decrease with increasing age, which may be accomplished by the excretion of mercury through the growth of feathers and as a dilution effect during growth. Selenium and mercury are inversely correlated in nestlings. The levels of zinc were similar for different nestling stages and between nestlings and adults in skuas and petrels. PMID- 11488241 TI - DNA adduct formation and DNA strand breaks in green-lipped mussels (Perna viridis) exposed to benzo[a]pyrene: dose- and time-dependent relationships. AB - Green-lipped mussels, Perna viridis, were exposed to 0, 0.3, 3 and 30 micrograms l-1 (nominal concentrations) B[a]P under laboratory conditions over a period of 24 days. Mussels were collected on day 0, 1, 3, 6, 12, 18 and 24, and the levels of DNA adducts and DNA strand breaks in their hepatopancreas tissues monitored. Mussels exposed to 0.3 and 3 micrograms l-1 B[a]P showed marked increases in strand breaks after 1 day of exposure. DNA strand break levels in these mussels remained high and significantly different from the control values until day 3 for the 0.3 microgram l-1 treatment group, and day 6 for the 3 micrograms l-1 treatment group. This was followed by a gradual reduction in strand breaks. After 12 days, the levels of both groups had returned to the same level as that of the control. No increase in DNA strand breaks was observable in mussels exposed to 30 micrograms l-1 B[a]P in the first 12 days of exposure, but a significant increase was observed from day 12 to day 24. Increasing B[a]P concentrations resulted in elevated DNA adduct levels after 3-6 days of exposure, but this pattern of dose related increase disappeared after 12 days. These results indicate that a better understanding of the complex interactions between exposure levels and durations is crucially important before DNA adduct levels and DNA strand breaks in P. viridis can be used as effective biomarkers for monitoring genotoxicants in marine waters. PMID- 11488242 TI - Polychlorinated biphenyls in marine sediments of Admiralty Bay, King George Island, Antarctica. PMID- 11488243 TI - Processes influencing mercury speciation and bioconcentration in the North Inlet Winyah Bay Estuary, South Carolina, USA. PMID- 11488244 TI - [Clinical research methods]. AB - The methodology of clinical research will be commented upon according to the components of the research design: a precise question, hypotheses derived from that which can be examined empirically, the biometrical estimation of the sample size and its practicability, the selection of control groups, measures for assessing psychopathological phenomena, analysing the data and preparing the findings for publication, further the time structure of the research project, and some aspects of the framework of clinical research: patient access, financing of the needed resources and ethical implications. PMID- 11488245 TI - [Concepts of vulnerability of psychiatric diseases]. AB - Despite intensive research in recent decades, the search for the aetiopathogenesis of psychiatric diseases is just as relevant as ever. In recent years, ideas of the aetiopathogenesis of psychotic diseases based on the concept of "vulnerability" in its diverse variations and developments have been gained increasingly ground. The publications of Zubin and associates above all have contributed to a "vulnerability model". According to this descriptive model, schizophrenic disturbances develop as a result of stimuli/irritants or stress factors under modulation of the social and physical environment as well as dependent on the premorbid personality. The increased vulnerability which gives rise to this is perceived as a threshold descensus of the individual towards stimuli/irritants (with a deficit counter--irritants impulses). Apart from that, multi-causality of vulnerability is assumed as the starting point, whereby there is the possibility of several therapy approaches. Moreover, contrary to the prevailing pessimistic view that schizophrenia is a process-type progressive disease, an episodic nature of schizophrenic psychoses is postulated with a prognosis that is indeed positive in the long term. In recent years, although often not explicitly stated, Zubin's concept of vulnerability was also indubitably subjected to various further developments (the vulnerability stress model by Nuechterlein and associates as also the integrative psycho-biological schizophrenia model by Ciompi may serve as examples). Worth mentioning are also various new concepts from system sciences (such as those from cybernetics, synergetics, the chaos theory, the communication theory, structure determinism etc.), which endeavour to clarify the problem of psychosis. The evaluation of central neurophysiological function deviations with schizophrenics and their relations has so far been oriented above all towards the vulnerability model of Zubin and Nuechterlein, which differentiates temporarily relatively stable trait markers with generally unaltered expressivity at the pre-, intra-, and post psychotic stages as well as episode markers and intermediate markers. However, specific factors contributing to the pathogenesis of schizophrenic disturbances have not as yet been found, in addition, there is still a multitude of methodological problems and distinctive features to fulfill the expectation of a comprehensive concept with which the whole complexity of the occurrence, the progress, and the outcome of psychoses can be explained. Reintroduction of the concept of vulnerability experienced, as Schmidt-Degenhardt put it, "a renaissance in use that appeared almost inflationary and a completely dubious popularisation ... without reference to its historical implications...". Consequently, a critical view of the use of this term would appear to be necessary. PMID- 11488246 TI - [Symptom-focussed drug therapy in borderline personality disorder]. AB - Borderline Personality Disorder (BPD) is a common and serious psychiatric illness. Psychopathological symptoms often are heterogenous and difficult to influence. Psychotherapy stands in the core of treatment efforts. However, during the treatment of these patients often situations occur that make the use of drug treatment necessary. Symptoms such as depression, impulsivity, or dissociation can be difficult to deal with in this population. There appears to be no treatment of choice for the disorder as a whole. Reviews so far have organized the literature by groups of pharmacological agents. In this review we give an overview over psychopathological symptoms in BPD, i.e. depression, affective dysregulation, aggression, impulsivity, dissociation, self-injuries, suicidality, psychotic symptoms, anxiety, obsessions and compulsions, and sleep disturbance. Effectivity of different drugs in influencing these symptoms is presented and significances of findings are used to assess the relevance of these findings. We conclude that certain symptoms such as depression or impulsivity can be successfully treated. Serotonin reuptake inhibitors play a decisive role in pharmacological treatment of Borderline Personality Disorder. Finally, recommendations for effective drug treatment of BPD symptoms are presented. Comorbid depression should be treated with Selective Serotonin Reuptake Inhibitors. These substances could also be shown to be effective in the treatment of impulsive and aggressive behavior. Intermittent psychotic symptoms are a target for atypical neuroleptics. Benzodiazepines are no longer viewed to be useful in the treatment of BPD. PMID- 11488247 TI - [Gestalt theory of V. v. Weizsacker from the viewpoint of the psychology emotions]. AB - The "Gestaltkreis" represents an important part in the psychosomatic theory discussion, which can be understood as an incomplete contribution towards a general theory of emotions. Especially the relationship between perception and motion requires further consideration. Instead of a causal attribution, cyclic complementaries are important. Furthermore different degrees of organisation within the body-scheme and the affect-apparatus are mentioned. PMID- 11488248 TI - [A psychiatrist under the Nazi dictatorship--Karl Bonhoeffer's commitment to racially and politically persecuted colleagues]. AB - Karl Bonhoeffer (1868-1948), professor of psychiatry at Breslau University and head of the psychiatric department of the Charite-hospital in Berlin, represented a view of personnel politics based solely on scientific and as personal competence. Anti-Semitism, often attributed to the faculty in Berlin during the Wilhelminian period and the Republic of Weimar, B. regarded as an utterly unsuitable criterion of personnel politics. Consequently, he had a considerable number of Jewish assistants as staff, about 30 altogether. In 1933, when massive persecution set in, quite a few of his students and assistants were persecuted for their political views. Some emigrated right away; others Bonhoeffer re employed in spite of their membership in illegal leftist parties. Even larger was the number of assistants who were racially persecuted. Bonhoeffer took advantage of loop holes in the anti-Semitic legalities, using complaints at the ministry or cleverly written applications to extend contracts even against high odds. In spite of his support, however, they eventually were fired and had to emigrate. They often found work in other countries because of his recommendations, though. For at least one assistant and one patient as well as their families, Bonhoeffer's commitment saved their lives. PMID- 11488249 TI - [Patterns of care and socio-professional categories derived from DIM (Department of Medical Information) data regarding MGEN psychiatric management]. AB - The influence of social factors (demographics, work statute, profession) on patterns of care like psychotropic drugs prescription, psychotherapeutic treatment or hospitalisation was studied on the population receiving care within the MGEN (a semiprivate health insurance system for the teaching related professions, the largest in France). This was possible by the use of a three year data bank (1992, 1993, 1994) where out and inpatients care were systematically recorded along some socio-demographics data in a rather detailed manner if provided within the system. The present study concerns 6,453 patients selected on the most frequent ICD 9 diagnoses as registered by the psychiatrist in charge of the patient: schizophrenia (acute, simple and persistent): 18.5% of the sample, manic-depressive psychosis 7.7%, and neurotic disorders (depressive disorders 38.3%, anxiety disorders 27% and mixed disorders 8.2%). For most of these diagnoses, hospitalization rates vary according the profession of the patient: teachers suffering from schizophrenia (simple) have higher rate of hospitalization than others professions; on the reverse university teachers and managers have the lowest rates of hospitalization for neurotic disorders. Duration of stay is rather identical within profession for psychotic disorders but again there are difference for neurotic disorders: blue collar workers, managers and university teachers have the lowest duration. Number of outpatients acts and provision of psychotherapeutic treatment either individual or group therapy show some variations between professions. Number of psychotropic drugs do not vary according profession but there is difference between men and women suffering from depressive disorders (2.97 for men, 2.72 for women). Neuroleptics are prescribed at the same rate but depot neuroleptics are prescribed mainly to blue collars workers, clerks and whose who are teaching technical matters; this applies only to men. These results are discussed according the context of the MGEN psychiatric health system. PMID- 11488250 TI - [Lethal catatonia: clinical aspects and therapeutic intervention. A review of the literature]. AB - Lethal catatonia continues to occur and represents a nonspecific syndrome associated with diverse organic as well as functional conditions. From this perspective, neuroleptic malignant syndrome may be conceptualized as a neuroleptic-induced toxic or iatrogenic form of organic lethal catatonia. Neuroleptics appear ineffective in the treatment of lethal catatonia and should be stopped whenever this disorder is suspected. Existing data suggest that ECT is a safe and effective treatment for lethal catatonia. ECT also appears effective in the treatment of neuroleptic malignant syndrome. PMID- 11488251 TI - [Anesthesia for electroconvulsive therapy: propofol versus thiopental]. AB - The anesthesia for sismotherapy is characterized by its briefness and repetitiveness, resulting in several imperatives: anesthesia of short duration, deep narcosis with muscular relaxation and ambulatory character. Thus anesthesic drugs should have a fast onset of action, in order to obtain a rapid and as alert as possible post anesthesia awakening. The objective of this study is to compare two anesthesic drugs: propofol versus thiopentone. We included in this study patients referred to our unit by the psychiatric service for sismotherapy, which was carried on under general anesthesia in the awakening room of the anesthesia department of Ibn Rochd University hospital. 7 of our patients received sismotherapy for schizophrenia, 2 for acute mania and 1 for suicidal depression. A total of 40 sessions of sismotherapy were analyzed, distributed in two groups: group I (n = 20): benefitted of a general anesthesia by thiopentone, the dose was 2 to 3 mg/kg; group II (n = 20): benefitted of general anesthesia by propofol, the dose was 1 to 1.5 mg/kg. Sismotherapy was carried out only once narcosis was considered as deep. To monitor our patients we used electrocardioscope and pulpe oxymeter. We evaluated the quality and especially the time of onset of anesthesia, its duration, the quality of narcosis, the degree of muscular relaxation, respiratory and cardiovascular parameters as well as side effect linked to anesthesia drugs and sismotherapy. Analysis of the results showed that the quality of anesthesia was excellent for both groups. The necessary dose for narcosis was 202 mg for thiopentone and 167 mg for propofol, time of onset of narcosis was 30 seconds for propofol and 45 seconds for thiopentone, anesthesia and the quality of muscular relaxation were considered deep for the two groups. Many authors showed that propofol is the most efficient agent in anesthesia for sismotherapy due to its brief delay of action and faster reversibility. As for thiopentone despite its convulsive properties and poor hemodynamic tolerance, it still is a good hypnotic in anesthesia for sismotherapy when administered at appropriate dose by slow injection. This is due on the one hand to easy administration, lesser incidence of side effects and on the other hand to brief duration of action and low cost. We conclude that thiopentone can be recommended in anesthesia for sismotherapy owing to good properties: deepness of anesthesia, good awakening, tolerance and lower cost. PMID- 11488252 TI - [Professional burnout: concept and critique of the burnout process, value of exploratory research in a prison milieu]. AB - The aim of this study is to suggest a critical analysis of burnout. In France, empirical studies devoted to the concept of burnout seem to be mainly focusing health on the professionals. However, a double limit persists. Firstly, the low number of current French publications. Secondly, the soundness of the theoretical models which consider the cognitive perception of subjects towards organizations. The analysis of theoretical issues confirms the considerable deficit of exploratory studies with correctional officers. Finally, we underline the interest to carry out an exploratory study applying to the subjects in order to release: sources of dissatisfaction related to work, their biopsychosocial consequences, in particular external constraints related to work, determinants of socialization and burnout. PMID- 11488253 TI - [Validation of a scale for responsibility (Salkovskis Responsibility Scale)]. AB - Appraisal of inflated responsibility for harm is the cornerstone of Salkovskis's cognitive theory for obsessive compulsive disorder. The aim of our study is to present the validation study of the French translation of the R scale. The present study compared 50 subjects with obsessive compulsive disorder, 37 patients suffering from social phobia and 183 control subjects on a responsibility questionnaire (R scale). The cognitive hypothesis of Obsessive Compulsive Disorder (OCD) specifies two levels of responsibility-related cognitions: responsibility assumptions (attitudes) and responsibility appraisals (interpretations). The R scale evaluates the responsibility assumptions. Such attitudes should reflect the more generalized tendency to assume responsibility in a given situation, particularly situations involving intrusions and doubts. It is possible that such assumptions may be less specific to OCD. The inclusion of social phobia subjects in the present study allows evaluation of the specificity of any findings to OCD. Patients were diagnosed and classified according DSM IV criteria. The control subjects were taken in the general population. No formal interview was conducted. The three groups were compared for sex, age and educational level. Before treatment, all the participants filled in the Responsibility Scale of Salkovskis (27 items), the Beck Depression Inventory (21 items), the Beck Anxiety Inventory and the Bouvard's Obsessive Compulsive Thoughts Checklist. The results indicate that the two anxious groups scored significantly higher than the control group on Beck Depression and Anxiety Inventories but no significant difference was observed between the two anxious groups. OCD patients scored significantly higher than both social phobic patients and control subjects on the Obsessive Compulsive Thoughts Checklist (OCTC). The social phobic group scored this checklist significantly higher than the control group. In sum, the three groups were different on obsessive compulsive thoughts. On the washing subscale of the Obsessive Compulsive Thoughts Checklist, the OCD patients differed significantly from the control group and the social phobia patients. No difference was observed between the social phobia subjects and the control group. On the two other subscales of the OCTC, the checking and the responsibility scales, the three groups were different: OCD patients scored significantly higher than both social phobic patients and control subjects; the social phobic patients scored higher than the control group. Results support the reliability (test retest) and the internal consistency of the questionnaire. Patients with obsessive compulsive disorder (OCD) and social phobia subjects had significantly elevated score on the total scale compared to control subjects. However social phobia patients did not differ from patients with OCD. So, the responsibility for harm, evaluated by the R-scale seems not to be specific of OCD. This finding does not support the results of two studies (28, 30). But these two studies compared OCD patients with an anxious group including panic disorder with agoraphobia, generalized anxiety disorder and social phobia. The correlations with a measure of OCD symptoms were higher than the correlations with anxiety and depression. Finally, the factor structure was only studied on the control group. The exploratory factor analysis indicates that the R scale is a two-dimensional scale, reflecting a need to prevent risks and the belief that one has power to harm. The first dimension is less specific to the pathology than the second. Only patients with OCD had significantly elevated score on the "need to prevent risks" compared to the non-clinical group. The two anxious groups differed on "the belief that one has power to harm" from the non-clinical group but social phobia patients did not differ from patients with OCD. In sum, the two subscales of the R scale did not discriminate OCD patients and social phobic subjects. Further research is needed to replicate the present findings and to confirm the two dimensions of the R scale. Overall, the results are consistent with the hypothesis that responsibility beliefs are important in the experience of obsessional problems. However, responsibility assumptions such as the belief that one has the power to harm are shared with social phobia. PMID- 11488254 TI - [Cross-sectional study of the prevalence of adjustment disorder with anxiety in general practice]. AB - Although Adjustment Disorder (AD) is considered a marginal diagnostic category by many clinicians and researchers, all the rare studies undertaken in the last decades indicate that the prevalence of this disorder is high in psychiatric settings, but has never been investigated in general practice. The purpose of this study was to evaluate the current prevalence of Adjustment Disorders With Anxiety (ADWA) in primary care settings and to describe the characteristics of the population, nature of the stressors and management of the disorder by General Practitioners (GPs). This French study involved 78 random liberal GPs, in 7 distinct regions (Paris, Lille, Bordeaux, Rouen, Dijon, Castres and Compiegne). GPs had to register all the consecutive attenders over 18 years old. For each physician, the registration period was over when 200 patients were registered, or 10 days of consultation were completed, or when 5 MINI had been performed. The average study period was 10 days per physician. At the first stage, they selected all the patients with psychological complaints, which were eventually associated to physical complaints. At the second stage, only the patients whose complaints were linked to a psychosocial stressor and without A1 and/or A2 DSM IV criteria for a Major Depressive Episode (MDE) were proposed the Mini International Neuropsychiatric Interview (MINI). The MINI is a brief structured clinical diagnostic interview that identifies the main axis-I DSM IV diagnoses in about 15 minutes. Before starting the study, all of the GPs participated in an intensive course on AD criteria recognition and were trained to use the MINI. The GPs registered a total of 7,759 consecutive patients. Twenty-two per cent (n = 1,719) of the patients reported psychological complaints, associated or not to physical complaints. Among them, 49% (n = 844) linked their complaints to identifiable psychosocial stressors. About half of the latter (n = 450) coded positive to A1 and/or A2 criteria for MDE. At the end, a total of 314 patients agreed to complete the MINI. Among the 1,719 patients with psychological complaints, the prevalence of ADWA eventually associated to other psychiatric disorders was 9.2%. The prevalence of "pure" ADWA was 4.5%. When considering the whole population of consecutive patients in primary care settings, the prevalence of pure ADWA was 1.0%. Patients suffering from pure ADWA were mostly women (66.7%), young patients (mean age: 42 years), with a professional activity. Patients had a psychiatric disorder history in 53.8% of the cases (mostly anxiety disorder). The main life events cited as being responsible for the disorder were work-associated problems (23.1%), followed by family illness (9.0%) and serious personal illness or accident (7.7%). The average duration of the disorder was 2.32 months. In 91% of the cases, GPs estimated that the patient required a pharmacological or psychological treatment. In most cases, they treated the patients with drug therapy (74.0%) associated with psychological support (counselling or psychotherapy, 76%). Anxiolytic agents were usually prescribed (64.9%), followed by antidepressants (10.8%) and hypnotics (8.1%). In conclusion, this first prevalence study of ADWA in general practice demonstrates that this disorder is frequent in primary care. It seems to be more present in patients who are of working age, especially women. ADWA would thus seem to preferentially affect active subjects. In most cases, GPs treat their patients with both psychological support and drug therapy. Anxiolytic is the elicited treatment of this disorder. PMID- 11488255 TI - [Role of the neurohypophysis in psychological stress]. AB - Effects of different psychological stimuli on oxytocin (OT) and vasopressin (AVP) secretion are reviewed in animals and in humans. The secretion of neuropituitary hormones is also discussed in various psychiatric diseases such an anorexia nervosa, bipolar disorder, schizophrenia and obsessive-compulsive disorder. AVP and OT are secreted into the hypophyseal portal circulation by neurons which project from the paraventricular nucleus to the external zone of the median eminence. AVP and OT-containing neurons in the suprachiasmatic and paraventricular nuclei project to limbic areas, including the hippocampus, the subiculum, the ventral nucleus of the amygdala and the nucleus of the diagonal band. Specific AVP receptors which are pharmacologically different from the pressor and antidiuretic AVP receptors have been found in the anterior pituitary. OT receptors have been identified in a variety of forebrain sites. The neurohypophyseal secretion is regulated by the cholinergic muscarinic, histaminergic and beta-adrenergic systems. Stress alters the secretion of one or more of the hypothalamic factors which interact at the pituitary to increase the secretion of ACTH. AVP and OT have been shown to modulate the effect of Corticotropin-Releasing Factor (CRF) on ACTH secretion and appear to play a key role in mediating the ACTH response to stress. Although AVP is a relatively weak secretagogue for ACTH, it markedly potentiates the activity of CRF both in vitro and in vivo. The role of OT is more complex. In vitro, OT stimulates ACTH release at high doses whereas in human it inhibits ACTH secretion at low doses. The type of stressor appear to determine the relative importance of these secretatogues in ACTH response. Several recent studies indicate that psychological stressors display a similar degree of variety of secretagogue release patterns as was found earlier for physical stressors. A bewildering array of technique produces a bewildering array of conclusions. In rats, OT may be an important secretagogue during a novel stimulus, whereas the role for AVP is less clear. Indeed two studies out of ten suggest a stimulating role for AVP. In response to frustration and submission, OT and AVP are secreted. Regarding social isolation, results are difficult to interpret and the role of AVP could be species-dependent. In contrast plasma OT levels do not change. After restraint, ACTH release is primarily mediated by the active increase of OT and AVP does not appear to play a role. When restraint is associated with moderate levels of physical components and during immobilisation, all two secretagogs are involved in the ACTH response. With fear, ACTH response appears to be driven by OT. In humans, one study indicates that high emotionality women increase plasma OT in response to uncontrollable noise. Various neuroendocrine dysregulations have been observed in psychiatric disease. Either an increase or a decrease of the hypothalamic pituitary-adrenal (HPA) function have been described in several illnesses. Effects of OT appear to be reciprocal to the effects of AVP. OT has been called the "amnestic" neuropeptide due to its capacity to attenuate memory consolidation and retrieval. AVP exhibits a central activating action on mood, memory and selective attention. Underweight patients with anorexia nervosa have abnormally high levels of centrally directed AVP and reduced OT levels. These modifications could enhance the retention of cognitive distortions of aversive consequences of eating. Patients with bipolar disorder show a biphasic secretion of AVP. Depressive episodes are associated with decreased vasopressinergic activity whereas manic episodes involve an increased release. AVP might be responsible for an increased catecholamine activity. In addition, lithium could act as an antagonist to AVP. In schizophrenic patients, studies using the apomorphine stimulation suggest increased oxytoninergic and decreased vasopressinergic functions. These findings are consistent with the beneficial role of AVP on schizophrenic symptoms noted in several trials. The increased OT could be responsible for "positive" symptomatology such as delusions and hallucinations. Obsessive compulsive disorder (OCD) includes a range of cognitive and behavioral disturbances that could be influenced by OT. In animals, several studies have emphasized the role of AVP in promoting repetitive grooming behaviors and maintaining conditioned response to aversive stimuli. In OCD patients, one study have reported that AVP/OT ratio was negatively correlated with symptom severity. However, an independent report found similar AVP concentrations in OC patients without a personal or family history of tic disorder and in normal subjects. Whether these modifications are only a consequence of the central disturbances or whether those peptides could participate in the pathogenesis of these affections remains to be elucidated. PMID- 11488256 TI - [Dynamic paradigm in psychopathology: "chaos theory", from physics to psychiatry]. AB - For the last thirty years, progress in the field of physics, known as "Chaos theory"--or more precisely: non-linear dynamical systems theory--has increased our understanding of complex systems dynamics. This framework's formalism is general enough to be applied in other domains, such as biology or psychology, where complex systems are the rule rather than the exception. Our goal is to show here that this framework can become a valuable tool in scientific fields such as neuroscience and psychiatry where objects possess natural time dependency (i.e. dynamical properties) and non-linear characteristics. The application of non linear dynamics concepts on these topics is more precise than a loose metaphor and can throw a new light on mental functioning and dysfunctioning. A class of neural networks (recurrent neural networks) constitutes an example of the implementation of the dynamical system concept and provides models of cognitive processes (15). The state of activity of the network is represented in its state space and the time evolution of this state is a trajectory in this space. After a period of time those networks settle on an equilibrium (a kind of attractor). The strength of connections between neurons define the number and relations between those attractors. The attractors of the network are usually interpreted as "mental representations". When an initial condition is imposed to the network, the evolution towards an attractor is considered as a model of information processing (27). This information processing is not defined in a symbolic manner but is a result of the interaction between distributed elements. Several properties of dynamical models can be used to define a way where the symbolic properties emerge from physical and dynamical properties (28) and thus they can be candidates for the definition of the emergence of mental properties on the basis of neuronal dynamics (42). Nevertheless, mental properties can also be considered as the result of an underlying dynamics without explicit mention of the neuronal one (47). In that case, dynamical tools can be used to elucidate the Freudian psychodynamics (34, 35). Recurrent neuronal networks have been used to propose interpretation of several mental dysfunctions (12). For example in the case of schizophrenia, it has been proposed that troubles in the cortical pruning during development (13) may cause a decrease in neural network storage ability and lead to the creation of spurious attractors. Those attractors do not correspond to stored memories and attract a large amount of initial conditions: they were thus associated to reality distorsion observed in schizophrenia (14). Nevertheless, the behavior of these models are too simple to be directly compared with real physiological data. In fact, equilibrium attractors are hardly met in biological dynamics. More complex behaviors (such as oscillations or chaos) should thus to be taken into account. The study of chaotic behavior have lead to the development of numerical methods devoted to the analysis of complex time series (17). These methods may be used to characterise the dynamical processes at the time-scales of both the cerebral dynamics and the clinical symptoms variations. The application of these methods to physiological signals have shown that complex behaviors are related to healthy states whereas simple dynamics are related to pathology (8). These studies have thus confirmed the notion of "dynamical disease" (20, 21) which denotes pathological conditions characterised by changes in physiological rhythms. Depression has been studied within this framework (25, 32) in order to define possible changes in brain electrical rhythms related to this trouble and its evolution. It has been shown that controls' brain dynamics is more complex than depressive one and that the recovery of a complex brain activity depends on the number of previous episodes. In the case of the symptoms time evolution, several studies have demonstrated that non-linear dynamical process may be involved in the recurrence of symptoms in troubles such as manic-depressive illness (9) or schizophrenia (51). These observations can contribute to more parcimonious interpretation of the time course of these illnesses than usual theories. In the search of a relationship between brain dynamics and mental troubles, it has been shown in three depressed patients an important correlation between the characteristics of brain dynamics and the intensity of depressive mood (49). This preliminary observation is in accordance with the emergence hypothesis according which changes in neuronal dynamics should be related to changes in mental processes. We reviewed here some theoretical and experimental results related to the use of "physical" dynamical theory in the field of psychopathology. It has been argued that these applications go beyond metaphor and that they are empirically founded. Nevertheless, these studies only constitute first steps on the way of a cautious development and definition of a "dynamical paradigm" in psychopathology. The introduction of concepts from dynamics such as complexity and dynamical changes (i.e. bifurcations) permits a new perspective on function and dysfunction of the mind/brain and the time evolution of symptoms. Moreover, it offers a ground for the hypothesis of the emergence of mental properties on the basis of neuronal dynamics (42). Since this theory can help to throw light on classical problems in psychopathology, we consider that a precise examination of both its theoretical and empirical consequences is requested to define its validity on this topic. PMID- 11488257 TI - [Body weight changes and psychotropic drug treatment: neuroleptics]. AB - Weight gain associated with neuroleptics or antipsychotic treatment is well known by psychiatrists, but is too rarely considered as justifying a specific treatment program. Overweight is a risk factor for somatic disorders and can have a negative influence on self-esteem and self-confidence. This can lead to poor observance, and relapse of psychotic symptoms. Some studies try to describe the weight fluctuations according to the different neuroleptics and taking into account other variables like treatment duration, age or sex. Mechanisms of weight gain are less studied, in spite of evidence that neuroleptics interact with receptors of dopamine, norepinephrine, serotonin, histamine and acetylcholin, all implicated in a way or another, in weight regulation. Antipsychotics, like clozapine and olanzapine, are more concerned with neuroendocrine and neurovegetative interactions, and are responsible for the most severe weight increases. Loxapine and molindone induce weight decreases, and these exceptions are difficult to explain. The paper discusses the clinical and the epidemiological data, and indicates the methodological problems for such studies. Some hypotheses about the pathophysiological aspects of this side effect are made, in regard to growing knowledge about the biological mechanisms of weight regulation. Some solutions for a better consideration and caretaking of patients with such problems or "at risk" treatment are proposed. PMID- 11488258 TI - [Prediction of treatment response in acute mania: controlled clinical trials with divalproex]. AB - OBJECTIVE: To determine predictive factors for response to mood stabilising treatment in manic episodes and to determine the mood stabilising properties of divalproex. METHODS: For predictive factors, 179 subjects in 3 parallel groups (divalproex, lithium, placebo) were evaluated over a period of 21 days by using structured interviews conducted by the clinician (SADS-C) and by nursing staff (ADRS). For the follow-on study, 372 stabilised patients were randomised to three groups: divalproex, lithium or placebo. RESULTS: The presence of depressive symptoms was associated with poor response to lithium, and patients with manic episodes with depressive symptoms or with rapid cycling exhibited good response to divalproex, while classical manic episodes showed good response to lithium and divalproex, and dysphoric or irritable manic episodes responded well to divalproex but not to lithium. A high number of both manic and depressive prior episodes is predictive of poor response to lithium and favourable response to divalproex. The effects of depressive and manic episodes appear to be independent and do not correlate with the duration of the illness or age at onset. Divalproex was superior to placebo in preventing all types of episodes, whether or not relapse was depressive or manic, and it was also superior to lithium in preventing depressive episodes. CONCLUSION: Specific features of the disease history and of the semiology of individual episodes help predict therapeutic response to mood stabilisers. PMID- 11488259 TI - [Value of fluoxetine in obsessive-compulsive disorder in the adult: review of the literature]. AB - Fluoxetine is a selective serotonin reuptake inhibitor (SSRI) with demonstrated efficacy in the treatment of major depressive episodes. Since 1985, it has been evaluated for the treatment of obsessive-compulsive disorder (OCD). The orbitofrontal cortex and caudate nucleus are cerebral structures believed to be involved in the pathogenesis of OCD, since hyperactivation of these territories in the basal state is corrected upon remission of symptoms induced by therapy with an SSRI or by behavioral psychotherapy. Furthermore, several studies have found abnormalities in serotoninergic transmission in the orbitofrontal cortex and SSRIs can increase serotonin release by desensitizing 5HTID autoreceptors. OCD is a severe, chronic psychiatric disorder frequently complicated by depressive episodes. Here we review the clinical trials of fluoxetine listed in the Medline and Embase computerized databases. Fluoxetine was found to be effective in OCD in all the published open-label studies as well as in placebo controlled trials with an effective dose range of 40 to 60 mg daily. Clinical evaluation was carried out by using specific scales such as the Y-BOCS or NIMH-OC and improvement was observed after several weeks of therapy. These studies comprising an extended phase showed that efficacy was maintained--for three years in the longest study--resulting in a higher percentage of responders relative to the treatment initiation phase. A comparison of fluoxetine and clomipramine showed comparable efficacy and a superior safety profile, both in terms of anticholinergic side effects and cardiotoxicity or overdosage. The relapse rate was similar with both drugs. In the four meta-analyses appearing in the databases, two studies found similar efficacy for clomipramine and fluoxetine. There are few studies which directly compare the different SSRIs, apart from a comparison of fluoxetine and sertraline showing that both drugs have similar efficacy. With clomipramine, the SSRIs represent the first-line treatment recommended by the experts, in association with behavioral therapy to improve and maintain the clinical response over the long term. The guidelines recommend an optimal fluoxetine dose of 40 to 60 mg daily with a minimum treatment duration of 1 to 2 years. Efficacy should not be evaluated before 8 weeks to allow for onset of the therapeutic effects. Fluoxetine was found to have a good safety profile in these studies and the adverse effects described (insomnia, headache, diminished libido) rarely led to discontinuation of the treatment. Adverse effects such as nervousness or insomnia at the start of therapy were predictors of a good response to fluoxetine, as were the presence of remissions, the absence of prior pharmacologic therapy and a high impulsiveness score. A long history of the disorder, severity of the symptoms, collection obsessions, washing compulsions, obsessional slowness and comorbidity with a schizotypic personality or vocal or motor tics were associated with a poorer response. Fluoxetine also alleviates collateral depressive symptoms by significantly reducing suicidal ideation and impulsiveness in OCD patients. Our study indicates that fluoxetine is effective and well tolerated in OCD, placing it among the first-line treatments recommended by consensus conference guidelines. PMID- 11488260 TI - [Apropos of atypical melancholia with Sustiva (efavirenz)]. AB - The treatment of HIV infection has changed dramatically in recent years as a result of the development of new drugs which allows a variety of multitherapy combinations more adapted to patients' needs and thereby improving compliance. Efavirenz is a non-nucleoside reverse transcriptase inhibitor. In addition to a potent antiretroviral activity, efavirenz is an easy-to-take drug with once-daily dosing and is usually well tolerated. Efavirenz, however, may induce psychic alterations which are variable and atypical in both their clinical presentation and severity. As early as the first days of treatment, efavirenz may provoke surprising phenomena such as nightmares, vivid dreams, hallucinations or illusions, and twilight states. Depersonalization and derealization episodes, personality alterations, stream of thought troubles and unusual thought contents, atypical depression and cognitive disorders have also been observed. These phenomena may occur either early or later on treatment. The prevalence of severe psychic disorders is less than 5%, but they are often responsible for harmful treatment discontinuations. Psychiatric side effects are heterogeneous and probably not related to pre-existing psychologic weakness. We do not have enough data to evaluate these side effects and their etiopathogeny. The drug could act directly on the central nervous system since it crosses the blood-brain barrier, on the serotoninergic and dopaminergic systems. Some authors have compared efavirenz-induced psychic effects to those associated with LSD and found structural similarities between the two molecules. However, the heterogeneity and low prevalence of the psychiatric side effects of efavirenz suggest and individual sensitivity. In order to improve patient care, a better clinical approach, neuropsychological evaluation, and functional brain imagery should be used to progress in the analysis and comprehension of these disorders. We discuss in this paper the case of Mister H. This HIV-infected person presented with two severe melancholic episodes associated with marked cognitive disorders which resisted two successive antidepressant treatments (viloxazine and citalopram, respectively) prescribed at effective doses and for sufficient time duration. Mister H. had no personal or family psychiatric antecedent. His psychic condition improved only when efavirenz was discontinued. However, drug discontinuation may not be an obligatory step to improve the patient's condition since antidepressant treatment has been found effective in some similar situations. Actually, each case should be discussed with the clinicians taking care of the patient. PMID- 11488261 TI - The US Public Health Service guideline for xenotransplantation: advances and limitations. PMID- 11488262 TI - Physician-system alignment: introductory overview. AB - The papers in this Special Supplement are based on research funded by the participating members of the joint Center for Health Management Research (CHMR) and Center for Organized Delivery Systems (CODS), and supported by the National Science Foundation under its Industry-University Cooperative Research Center Program. This 3-year research initiative from 1996 through 1999 involved 69 physician organizations (primarily organized medical groups as opposed to IPAs) associated with 14 organized delivery systems. The groups ranged in size from three to 958 with an average size of 76.4 and a median size of 25.0. Comparisons of the study groups with United States physician groups overall are shown in Table 1. The study groups are larger and more likely to be multispecialty than all groups in the United States. The organized delivery systems range in size from one hospital to 80 hospitals with an average of 21 hospitals per system and a median of 11 hospitals per system. They average 4.6 affiliated medical groups with a range from one to 23. The organized delivery systems range in total revenues in 1998 from $340 million to $6.2 billion with an average of $2.1 billion. All the study systems are not-for-profit. Most are located in single market areas, but several are located in multiple markets. For the most part, they represent some of the larger most experienced organized delivery systems in the country. Among the primary objectives of the study was to identify the factors most strongly associated with physician alignment with the health care system and the consequences for the implementation of evidence-based care management practices. The study was also designed to identify the barriers and facilitators to achieving such alignment and its consequences. PMID- 11488263 TI - The ties that bind: interorganizational linkages and physician-system alignment. AB - OBJECTIVES: To examine the association between the degree of alignment between physicians and health care systems, and interorganizational linkages between physician groups and health care systems. METHODS: The study used a cross sectional, comparative analysis using a sample of 1,279 physicians practicing in loosely affiliated arrangements and 1,781 physicians in 61 groups closely affiliated with 14 vertically integrated health systems. Measures of physician alignment were based on multiitem scales validated in previous studies and derived from surveys sent to individual physicians. Measures of interorganizational linkages were specified at the institutional, administrative, and technical core levels of the physician group and were developed from surveys sent to the administrator of each of the 61 physician groups in the sample. Two stage Heckman models with fixed effects adjustments in the second stage were used to correct for sample selection and clustering respectively. RESULTS: After accounting for sample selection, fixed effects, and group and individual controls, physicians in groups with more valued practice service linkages display consistently higher alignment with systems than physicians in groups that have fewer such linkages. Results also suggest that centralized administrative control lowers physician-system alignment for selected measures of alignment. Governance interlocks exhibited only weak associations with alignment. CONCLUSIONS: Our findings suggest that alignment generally follows resource exchanges that promote value-added contributions to physicians and physician groups while preserving control and authority within the group. PMID- 11488264 TI - Risk assumption and physician alignment with health care organizations. AB - OBJECTIVES: To examine the association between risk assumption by individual physicians and physician groups and the degree of alignment between physicians and health care systems. METHODS: A cross sectional comparative analysis using a sample of 1,279 physicians practicing in loosely affiliated arrangements and 1,781 physicians in 61 groups closely affiliated with 14 vertically integrated health systems. Measures of physician alignment were based on multiitem scales validated in previous studies and derived from surveys sent to individual physicians. Measures of risk assumption were developed from surveys sent to the administrator of each of the 61 physician groups in the sample and to physicians affiliated with these groups. Two stage Heckman models with fixed effects adjustments in the second stage were used to correct for sample selection and clustering respectively. RESULTS: After accounting for selection, fixed effects, and group and individual controls, physicians in groups with larger proportional revenue from managed care displayed greater normative commitment and system loyalty than physicians in groups with lower proportional managed care revenue. Individual-level managed care risk was also positively related to both normative commitment and group behavioral commitment to the system. Physicians in groups with larger physician equity positions expressed lower levels of normative commitment to the system. Physician productivity compensation was negatively related to all measures of alignment. Finally, group emphasis on individually based incentives for staff physicians was negatively related to system identification. CONCLUSIONS: Our findings suggest that organizations must balance individually-based risk schemes with those that emphasize the performance of the group and the system to achieve long-term goals of loyalty, identification, and commitment to the system. PMID- 11488265 TI - Implementing evidence-based medicine: the role of market pressures, compensation incentives, and culture in physician organizations. AB - OBJECTIVES: To assess the extent to which market pressures, compensation incentives, and physician medical group culture are associated with the use of evidence-based medicine practices in physician organizations. METHODS: Cross sectional exploratory study of 56 medical groups affiliated with 15 integrated health systems from across the United States, involving 1,797 physician respondents. Larger medical groups and multispecialty groups were overrepresented compared with the United States as a whole. Data are from two sources: (1) surveys of physicians assessing the culture of the medical groups in which they work, and (2) surveys of medical directors and other managerial key informants pertaining to care management practices, compensation methods, and the management and governance of the medical groups. Physician-level data were aggregated to the group level to attain measures of group culture and then merged with the data regarding care management, incentives, and management and governance. Stepwise multiple regression was used to examine the study hypotheses. RESULTS: As hypothesized, the number of different types of compensation incentives used (cost containment, productivity, quality) was positively associated with the comprehensiveness of care management practices. The degree of salary control (ie, market-based salary grades and ranges versus the use of bookings or fees and individual negotiation) was also positively associated with the deployment of care management practices. As hypothesized, market pressures in the form of percentages of health maintenance and preferred provider organization patients seen were generally positively associated with the use of care management practices. Organizational culture had no association except that a patient centered culture in combination with a greater number of different types of compensation incentives used was positively associated with greater use of care management practices. CONCLUSIONS: Both compensation incentives and managed care market pressures were significantly associated with the use of evidence-based care management practices. The lack of association for culture may be due to the relatively amorphous nature of most physician organizations at this point. PMID- 11488266 TI - Factors associated with physician involvement in care management. AB - BACKGROUND: Enthusiasm for the concept of care management (CM) has led to unprecedented growth in the number of guidelines and protocols, but provider organizations have struggled to enlist the active support and participation of physicians in CM activities. OBJECTIVES: To empirically examine the factors influencing physician participation in and attitudes toward CM activities. METHODS: Data on 1,514 physicians were used to predict physician attitudes toward CM and their perceptions of group CM behaviors. Dependent variables were modeled using two-stage Heckman selection bias models with fixed effects corrections. Independent predictors included physician- and group-level controls as well as six potential CM participation and attitude facilitators. RESULTS: Physician participation in the implementation phase of CM activities was positively related to participation and attitude. However, physician participation in the development phase may be negatively related to later participation in CM activities. Management involvement in development phase has mixed effects (positive or no effect), but their involvement in the implementation phase was somewhat negatively related to CM participation and attitude. Financial incentives for participation in CM activities and presence of a useful management information system also appeared to be positively related to attitude and participation. CONCLUSIONS: Appropriate physician and management involvement, as well as financial incentives and useful management information systems may facilitate physician participation in CM activities. Physician involvement in implementation of CM practices appears to be important, whereas their involvement in the development phase may be negatively related to later attitudes and participation. The findings call for a more in-depth understanding of the timing of physician input in CM activities. PMID- 11488267 TI - Physician commitment to organized delivery systems. AB - BACKGROUND: Health care systems have developed many types of contracting vehicles with physicians. The immediate aim of these vehicles has been to foster physician commitment and alignment to the system. The ultimate aim of these vehicles has been to garner managed care contracts, reduce costs, and improve quality. To date, most of these vehicles have failed to improve physician commitment. This may be one reason why the ultimate outcomes have not been observed. Consequently, systems are experimenting with new methods to partner with physicians. One new method is to segment physicians into tightly linked and loosely linked strategic alliances and devote different levels of resources and attention to each. OBJECTIVES: This study evaluates whether the segmentation of physicians into tightly linked versus loosely linked strategic alliances improves the commitment of physicians to the system. The study then investigates which constituent elements of the tightly linked strategic alliances exhibit the greatest association with commitment. DESIGNS AND SUBJECTS: The study uses a cross sectional design and survey data drawn from 1,965 physicians affiliated with 14 health care systems around the country. Tightly linked physicians typically practiced in hospital-sponsored group practices, whereas loosely linked physicians typically used the system's hospitals as their primary site of inpatient practice. MEASURES: Commitment is measured by seven different scales drawn from the literature on organizational commitment, loyalty, and identification. Some of the scales refer to physician attitudes, whereas others describe physician behaviors. The literature suggests that commitment is associated with both instrumental/utilitarian considerations (eg, older age, tenure with system, admissions to system, receipt of a stipend, etc.) as well as administrative involvement/participation considerations (eg, decision-making roles). A series of physician background and practice characteristics are used here to model these two types of factors. RESULTS: The study finds small but significant differences in commitment between physicians in tightly linked versus loosely linked alliances. Multivariate analyses suggest that instrumental/utilitarian factors (eg, age, receipt of stipend, percent of admissions to the system) may exhibit stronger associations with commitment than the physician's administrative involvement in the organization. CONCLUSIONS: To the degree that physician commitment is possible, systems should appeal to physicians' calculative motivations using extrinsic rewards rather than normative involvement in the organization. PMID- 11488268 TI - Physician-system relationships: stumbling blocks and promising practices. AB - OBJECTIVES: To identify the barriers, facilitators, and potential better practices to achieving physician-system alignment. METHODS: Interviews using a semi-structured, open-ended protocol were conducted during a total of 18 site visits, each usually 2 days in length, covering multiple topics of physician group-system alignment. Interviews were conducted with members of the target physician group, key leaders of the health care system, and representatives of physicians not in the target group. The summary of the interviews for each of the site visits was analyzed to determine barriers, facilitators, and better practices for achieving more effective relationships between physician groups and health care systems. RESULTS: A number of barriers to more effective relationships between physician groups and health systems were identified. Barriers related to environment, culture, and information systems were most prevalent. Other major general areas of barriers encountered were physician leadership, group-system relationship, compensation and productivity, care management practices, group strategy, and accountability. Examples of practices that may help to resolve some of these issues were also identified. CONCLUSIONS: Physician-system relationships can and do cause problems for improving health care. The evidence from the conducted site visits suggests that specific strategies may help improve these relationships but more research is needed in order assess the actual impact of these strategies. PMID- 11488269 TI - Kinetics of VO(2) with very high intensity exercise. PMID- 11488270 TI - USAN Council. List No. 439. New names. Laronidase. PMID- 11488271 TI - If you had a magic wand. . . PMID- 11488272 TI - Training could prevent deaths due to rodent gavage procedure. PMID- 11488273 TI - Rodent gavage technique concerns: avoiding excess mortality. PMID- 11488274 TI - An ethicist's view on the inclusion of rats, mice and birds under the Animal Welfare Act. PMID- 11488275 TI - Feedback on alternative solutions to USDA inspection of rats, mice and birds. PMID- 11488276 TI - Sustained downgaze in coma after cardiac arrest. PMID- 11488277 TI - Prion protein gene polymorphism and Alzheimer's disease: one modulatory trait of cognitive decline? PMID- 11488278 TI - Complex musical hallucinosis in a professional musician with a left subcortical haemorrhage. PMID- 11488279 TI - Cochlear implantation in a profoundly deaf patient with MELAS syndrome. PMID- 11488280 TI - Lead poisoning from complementary and alternative medicine in multiple sclerosis. PMID- 11488281 TI - Anti-GQ1b IgG antibody syndrome without ophthalmoplegia: clinical and immunological features. PMID- 11488282 TI - Post-operative microalbuminuria in congenital heart failure. PMID- 11488283 TI - Unusual variety of eustachian valve. PMID- 11488284 TI - Differential expression of 3 types of NOS in rat myocardial infarction. PMID- 11488285 TI - Reversed flow in internal mammary artery conduit and vertebral artery with left subclavian artery occlusion causing angina and vertigo. The coronary--subclavian steal syndrome. PMID- 11488286 TI - A new embryonic linkage between chromosome 22q11 deletion and a right ductus from a right aortic arch in a neonate with DiGeorge syndrome. PMID- 11488287 TI - Repeated coronary emboli in a young female with mild mitral stenosis in sinus rhythm. PMID- 11488288 TI - Reversible acute global left ventricular dysfunction in a patient with autosomal recessive dystrophic epidermolysis bullosa. PMID- 11488289 TI - Myxoma producing right-sided inflow and outflow obstruction. PMID- 11488290 TI - Vanishing cardiac tumor. PMID- 11488291 TI - Stuck with the bill. PMID- 11488292 TI - 1Q[3a]. Coverage for complementary & alternative medicine? PMID- 11488293 TI - Nova Award winners 2001. PMID- 11488294 TI - Factor variability of the Center for Epidemiological Studies Depression Scale (CES-D) among urban Latinos. AB - Establishing comparable measurement properties across different populations or in different population subgroups is a crucial yet often neglected step in instrument development. Failure to have comparable factor structures across groups makes any comparison between groups suspect. Previous analyses of the measurement structure of the Center for Epidemiologic Studies Depression Scale (CES-D) in diverse racial/ethnic populations have resulted in conflicting results. In the present analysis, data from three studies of urban Latinos (N = 1,403) were analyzed using structural equation modeling techniques to (1) fit the original four-factor solution separately in men and women; (2) evaluate configural and metric invariance between men and women; and (3) evaluate the mediating effects of age and acculturation on the fit of this model to the data. Results indicated that the four-factor model proposed by Radloff provided adequate fit to the data for Latina women when age and acculturation were included in the model. The four-factor model did not fit the data for Latino men; thus tests of configural and metric invariance across these two groups failed. We conclude that the CES-D may not measure the same constructs in Latino men and women and that further evaluation of the use of this measure in diverse populations is needed. Additionally, prior to comparison with other groups in which the four-factor solution is observed, the effects of age and acculturation should be controlled in Latinas. PMID- 11488295 TI - Multiple sclerosis: more than inflammation and demyelination. PMID- 11488296 TI - Functional recovery after spinal cord injury: basic science meets clinic. PMID- 11488297 TI - SNAREdinburgh: the molecular mechanisms of exocytosis and endocytosis. PMID- 11488298 TI - Changes in serum and hepatic cholesterol in lead-induced liver hyperplasia. AB - Lead nitrate when injected intravenously as a single dose to male Wistar rats causes a strong hepatic proliferative response followed by reabsorption of excess tissue within 10-14 days. The rate of cell proliferation in this hyperplastic model was positively correlated with hepatic de novo synthesis of cholesterol, stimulation of the hexose monophosphate shunt pathway of glucose metabolism and with alterations in serum lipoproteins. PMID- 11488299 TI - Lung-volume reduction. PMID- 11488300 TI - The symptomatic adolescent five years later: he didn't grow out of it. PMID- 11488301 TI - Transfer of phospholipids from fat body to lipophorin in Rhodnius prolixus. AB - 32P-Labeled fat bodies (32P-fat bodies) of Rhodnius prolixus females were incubated in the presence of non radioactive purified lipophorin and the release of radioactivity to the medium was analysed to answer the question of whether lipophorin is a reusable shuttle for phospholipids. The radioactivity found in the medium was associated with lipophorin phospholipids. When the 32P-fat bodies were incubated in the absence of lipophorin, only a small amount of radioactivity was released and it was not associated with lipophorin, indicating that there was no release of pre-labeled 32P-lipophorin by the tissue. Analysis of 32P phospholipids transferred from fat bodies to the lipophorin particles by thin layer chromatography revealed a predominance of phosphatidylethanolamine and phosphatidylcholine, with minor amounts of phosphatidylserine, phosphatidylinositol, and sphingomyelin. The transfer of phospholipids to lipophorin was linear with time up to 45 min and the process was inhibited at low temperature and by the metabolic inhibitors azide and fluoride. The transfer of phospholipids from the fat bodies to lipophorin was saturable with respect to the concentration of lipophorin, which was half-maximal at about 8 mg/ml. A directional movement of phospholipids from the fat body to lipophorin was observed. The net gain of phospholipids in 2 h of incubation with fat body was 8.54 nmol per insect, which corresponds to 6.69% of increase in the lipophorin phospholipid content. The rate of 32P-phospholipid transfer from fat body to lipophorin particles varied during the days after a blood meal increasing up to day 10 and then decreasing in parallel with the process of oogenesis. PMID- 11488302 TI - Loading of lipophorin particles with phospholipids at the midgut of Rhodnius prolixus. AB - 32P-Labelled midguts (32P-midguts) of Rhodnius prolixus females were incubated in the presence of nonradioactive purified lipophorin and the release of radioactivity to the medium was analysed. The radioactivity found in the medium was associated with lipophorin phospholipids. When the 32P-midguts were incubated in the absence of lipophorin, no 32P-phospholipids were found in the medium. Comparative analysis by thin-layer chromatography of 32P-phospholipids derived from metabolically labelled 32P-midgut or lipophorin particles after incubation with 32P-midgut showed some differences, revealing a possible selectivity in the process of phospholipids transfer. The transfer of phospholipids to lipophorin was linear with time up to 45 min, was saturable with respect to the concentration of lipophorin, and was half-maximal at about 5 mg/ml. The binding of 32P-lipophorin to the midgut at 0 degrees C reached the equilibrium at about 1 h of incubation. The binding of 32P-lipophorin was inhibited by an excess of nonradioactive lipophorin, which suggests a specific receptor for lipophorin. The capacity of midguts and fat bodies to transfer phospholipids to lipophorin varied during the days following the meal. When lipophorin enzymatically depleted of phospholipids by treatment with phospholipase A2 was incubated with 32P-midguts, the same amount of phospholipids was transferred, indicating a net gain of phospholipids by the particle. PMID- 11488303 TI - Characterization and immunocytochemical localization of lipophorin binding sites in the oocytes of Rhodnius prolixus. AB - Purified lipophorin, metabolically labelled with 32P exclusively in the phospholipid moiety, was used to study the process of phospholipid delivery to the oocyte. The kinetics of phospholipid transfer "in vitro," from lipophorin to the oocytes, was linear at least up to 4 h and was impaired by low temperature. A net transfer of phospholipids from lipophorin particles to the oocytes was observed. The rate of phospholipid uptake was dependent on the concentration of lipophorin in the medium and was shown to be a saturable process. The addition of a molar excess of purified unlabelled lipophorin to the culture medium resulted in a substantial decrease in the transfer of [32P]phospholipids, but no reduction occurred in the presence of a molar excess of albumin. The lipophorin binding sites were localized in the oocytes by immunogold techniques using two different protocols for oocyte fixation. Strong labelling was observed especially at the microvilli. No labelling was detected in the yolk granules. PMID- 11488304 TI - Pulmonary ventilation imaged by magnetic resonance: at the doorstep of clinical application. AB - Over the past few years, magnetic resonance imaging (MRI) has emerged as an important instrument for functional ventilation imaging. The aim of this review is to summarize established clinical methods and emerging techniques for research and clinical arenas. Before the advent of MRI, chest radiography and computed tomography (CT) dominated morphological lung imaging, while functional ventilation imaging was accomplished with scintigraphy. Initially, MRI was not used for morphological lung imaging often, due to technical and physical limitations. However, recent developments have considerably improved anatomical MRI, as well as advanced new techniques in functional ventilation imaging, such as inhaled contrast aerosols, oxygen, hyperpolarized noble gases (Helium-3, Xenon 129), and fluorinated gases (sulphur-hexafluoride). Straightforward images demonstrating homogeneity of ventilation and determining ventilated lung volumes can be obtained. Furthermore, new image-derived functional parameters are measurable, such as airspace size, regional oxygen partial pressure, and analysis of ventilation distribution and ventilation/perfusion ratios. There are several advantages to using MRI: lack of radiation, high spatial and temporal resolution and a broad range of functional information. The MRI technique applied in patients with chronic obstructive pulmonary disease, emphysema, cystic fibrosis, asthma, and bronchiolitis obliterans, may yield a higher sensitivity in the detection of ventilation defects than ventilation scintigraphy, CT or standard pulmonary function tests. The next step will be to define the threshold between physiological variation and pathological defects. Using complementary strategies, radiologists will have the tools to characterize the impairment of lung function and to improve specificity. PMID- 11488305 TI - Urban air pollution and chronic obstructive pulmonary disease: a review. AB - There is consistency in the findings that relate the acute increases in urban air pollution (mainly the particulate matter) and the short-term health effects (i.e. mortality and hospital admissions) on patients suffering from chronic obstructive pulmonary disease (COPD). Beyond the acute effects, a relevant public health and scientific question is to what extent chronic exposure to air pollution is related with lung function impairment and development of COPD. The few cross sectional studies showed an increase of self-reported diagnosis of chronic bronchitis and emphysema, breathlessness and mucus hypersecretion and lower levels of lung function in the more polluted areas. The two cohort studies in children have found a retardation of pulmonary function growth, and the only cohort in adults a faster decline of lung function. Given the intrinsic limitations of cross-sectional studies to separate the timing between exposure and effects, and the weaknesses of the cohorts (ie. few areas, short follow-up, only one retest, lack of individual weighting of air pollution measure) there is a need for new prospective studies. These should be performed in a large number of geographical areas and with a long follow-up to asses the impact of long-term air pollution on lung function growth and decline, and on incidence, duration, severity and survival of chronic obstructive pulmonary disease. PMID- 11488306 TI - Ventilator-associated pneumonia. European Task Force on ventilator-associated pneumonia. PMID- 11488307 TI - Human leukocyte antigen genes may outweigh racial background when generating a specific immune response in sarcoidosis. AB - Presented is a sarcoidosis patient of African origin, who was adopted at an early age and grew up in Sweden. This patient displayed an immune response identical to that previously reported in human leukocyte antigen (HLA)-DR17 positive Caucasian sarcoidosis patients in Scandinavia, with T-cell receptor AV2S3+ T-cells accumulating in the lungs. HLA typing also established that she was DR17 positive, which is a rare HLA type for individuals of African origin. To the authors' knowledge, this specific immune response has not previously been reported in patients of African origin. Moreover, the clinical manifestations of sarcoidosis were similar to those known to be strongly linked to HLA-DR17 in Scandinavians, i.e. with Lofgren's syndrome. The case presented here suggests certain human leukocyte antigen genes to be strongly linked to specific immune responses that are identical irrespective of the racial background. If such an immune response were important for the subsequent clinical manifestations, this case would argue for the importance of human leukocyte antigen genes in the genetic predisposition to sarcoidosis. PMID- 11488308 TI - Interleukin-12 as successful adjuvant in tuberculosis treatment. AB - Interleukin-12 (IL-12) proved to be an effective and successful adjuvant to a standard antituberculotic medication in a patient suffering from progressive clinical tuberculosis (TB). IL-12 is a potent enhancer of interferon-gamma production which is necessary for killing intracellular bacteria like mycobacteria. This patient's TB was progressive, although sensitivity to first line antituberculotics was proven and medication was given as directly observed therapy over more than 8 months. The 3-month adjuvant therapy with IL-12 significantly and convincingly improved results. It is believed that this case, the first in the literature to describe adjuvant interleukin-12 therapy in tuberculosis, strongly encourages the study of adjuvant interleukin-12 therapy on a more systematic basis. PMID- 11488309 TI - Allergic bronchopulmonary aspergillosis in cystic fibrosis. PMID- 11488310 TI - Does theophylline really improve acute mountain sickness? PMID- 11488311 TI - Not all laboratories are alike. PMID- 11488312 TI - Daily respiratory mortality and PM10 pollution in Mexico City. PMID- 11488313 TI - The search for the Holy Grail and the upper airway resistance syndrome. New data are welcome. PMID- 11488314 TI - Sleep and daytime sleepiness in upper airway resistance syndrome compared to obstructive sleep apnoea syndrome. AB - This study has investigated differences in the nocturnal sleep and daytime sleepiness among patients with obstructive sleep apnoea syndrome (OSAS), upper airway resistance (UARS), sleep hypopnoea syndrome, and normal control subjects, using sleep scoring and spectral activity analysis of the electroencephalogram (EEG). Twelve nonobese males with UARS aged 30-60 yrs were recruited. These subjects were strictly matched for age and body mass index with twelve OSAS patients, 12 sleep hypopnoea syndrome patients, and 12 normal controls, all male. Daytime sleepiness was evaluated using the Epworth Sleepiness Scale (ESS) and the Multiple Sleep Latency Test (MSLT). The macrostructure of sleep was determined using international criteria and spectral analysis of the sleep EEG was obtained from a central lead. The sleep macrostructure of OSAS and UARS patients was significantly different from that of controls. These patients were also sleepier during the daytime than controls. Complaints of tiredness and daytime sleepiness, ESS and MSLT scores were similar in the different patient groups. Mild dysmorphia was present in all three patient groups. However, nocturnal sleep was significantly different among the different groups. OSAS patients had significantly more awake time during sleep than the UARS patients. The spectral activity of the total sleep time of the patient groups also differed significantly from that of controls. When the sleep spectral activity of UARS and OSAS patients were compared, OSAS patients had less slow wave sleep activity than UARS patients. UARS patients had a significantly higher absolute power in the 7-9 Hz bandwidth than OSAS patients. The absolute delta power over the different sleep cycles was also different between controls and patients, and between UARS and OSAS patients. There are clear differences in the macrostructure and spectral activity of sleep between upper airway resistance and obstructive sleep apnoea syndrome patients, demonstrated by differences in the cortical activity recorded in the central lead during sleep. Despite these nocturnal sleep differences, the tests of subjective daytime sleepiness are not significantly different. PMID- 11488315 TI - Outcome of COPD patients with mild daytime hypoxaemia with or without sleep related oxygen desaturation. AB - The aim of the present study was to compare the evolution of pulmonary haemodynamics and of arterial blood gases in chronic obstructive pulmonary disease (COPD) patients with mild-to-moderate hypoxaemia, with or without sleep related oxygen desaturation. COPD patients with daytime arterial oxygen partial pressure in the range 56-69 mmHg were included prospectively. Sleep-related oxygen desaturation was defined as spending > or = 30% of the nocturnal recording time with arterial oxygen saturation <90%. From the 64 patients included, 35 were desaturators (group 1) and 29 were nondesaturators (group 2). At baseline (t0), patients with sleep-related desaturation had a significantly higher daytime (mean +/- SD) arterial carbon dioxide partial pressure (Pa,CO2) (44.9 +/- 4.9 mmHg versus 41.0 +/- 4.1 mmHg, p=0.001) whereas mean pulmonary artery pressure (mPAP) was similar in the two groups. After 2 yrs (t2) of follow-up, 22 desaturators and 14 nondesaturators could be re-evaluated, including pulmonary haemodynamic measurements. None of the nondesaturator patients became desaturators at t2. The difference between the two groups in terms of daytime Pa,CO2 was still present at t2. The mean changes in mPAP from t0 to t2 were similar between the two groups, as were the rates of death or requirement for long-term oxygen therapy (American Thoracic Society criteria) during follow-up of up to 6 yrs. The presence of sleep related oxygen desaturation is not a transitional state before the worsening of daytime arterial blood gases, but is a characteristic of some chronic obstructive pulmonary disease patients who have a higher daytime arterial carbon dioxide partial pressure. Such isolated nocturnal hypoxaemia or sleep-related worsening of moderate daytime hypoxaemia does not appear to favour the development of pulmonary hypertension, nor to lead to worsening of daytime blood gases. PMID- 11488316 TI - High or standard initial dose of budesonide to control mild-to-moderate asthma? AB - Guidelines on the use of inhaled steroids in asthma advocate that the daily dose should be chosen according to the severity of the disease. However, the question of the optimal starting dose remains to be properly addressed, as does the issue of the adjustment in dose required for a given patient. Whether a high initial dose of budesonide (800 microg b.i.d) was more efficacious than a standard dose (200 microg b.i.d) in controlling mild-to-moderate asthma was investigated, and whether the dose could be decreased, based on peak expiratory flow (PEF), symptom score, beta2-agonist use in a double-blind, randomized, parallel-group 18-week study. One-hundred and sixty-nine patients (mean age 38 yrs, mean forced expiratory volume in one second 74% predicted) were enrolled. No difference was detected between the two groups in improvement in morning PEF (+61 L x min(-1) in the high-dose group, +60 L x min(-1) in the standard-dose group by 16 weeks). Morning and evening PEF values stabilized before the end of the first 4 weeks. No difference between groups was observed in symptom score, beta2-agonist use, number of exacerbation per interval and the best forced expiratory volume in one second achieved. The proportion of subjects being able to decrease the doses of budesonide was similar in both treatment strategies. It is concluded that both high and standard initial doses are equally effective in controlling symptoms and improving lung function in mild-to-moderate asthma. PMID- 11488317 TI - Comparison of the acute effects of salbutamol and terbutaline on heart rate variability in adult asthmatic patients. AB - This study investigated the effects of beta2-adrenergic agonist therapy on heart rate variability (HRV) in adult asthmatic patients by using frequency domain measures of HRV. A randomized crossover design was used. Twenty adult patients with asthma were studied. All patients showed a mild-to-moderate decrease in baseline forced expiratory volume in one second. Any diseases that might have influenced the autonomic function were excluded. All patients had a complete physical examination and medical history that revealed no cardiovascular disease or medication. The study used 200 microg inhaled salbutamol and 500 microg inhaled terbutaline. HRV analysis was performed for each 5-min segment, 5 min before inhalation of the study drug and 5, 10, 15, 20, 25 and 30 min after inhalation. Total power (TP: <0.40 Hz), high-frequency power (HF: 0.15-0.40 Hz), low-frequency power (LF: 0.04-0.15 Hz) and LF/HF ratio were calculated. The LF and LF/HF ratio increased and TP decreased at 5, 10, 15 and 20 min after the salbutamol and the terbutaline inhalation, HF did not change significantly after the salbutamol and terbutaline inhalation. Acute salbutamol and terbutaline inhalation produce similar effects on heart rate variability and increase sympathetic modulation in the cardiac autonomic activity. PMID- 11488318 TI - Increased nitric oxide production by neutrophils in bronchial asthma. AB - This study was designed to assess the production of nitric oxide (NO) by neutrophils in bronchial asthma. Thirty asthmatic patients (ten each of mild, moderate and severe asthma) and ten healthy controls were included in the study. Neutrophils from peripheral venous blood were stimulated with latex, and production of nitrite (an NO metabolise) and L-citrulline (a co-product of NO) was studied. It was postulated that peripheral blood neutrophils, being in a primed or activated state in asthma, would reflect the changes occurring in bronchial tree neutrophils. Nitrite and L-citrulline production by neutrophils was significantly higher in asthmatics (p < 0.001) and increased with disease severity. A strong negative correlation was observed between peak expiratory flow and both nitrite (r = -0.87, p < 0.001) and L-citrulline (r = -0.88, p < 0.001) production. It is concluded that nitric oxide production by neutrophils is increased in bronchial asthma and can possibly contribute to airway narrowing and disease severity. PMID- 11488319 TI - Reduced late asthmatic response by repeated low-dose allergen exposure. AB - Allergic asthmatic individuals are often exposed to low-doses of allergen in their everyday life. Extended exposure to allergen has lead to down-regulation of the allergic process in cell systems and in animal models. The aim of this study was to evaluate whether any such inhibitory mechanism of allergic responses can be seen in man in vivo. Patients with mild asthma were repeatedly and double blindly exposed to 25% of the individual dose of allergen that caused an early (EAR) and late asthmatic reaction (LAR). One day after the low-dose allergen or placebo exposure periods, the same individual was given a high-dose allergen challenge. Sputum and blood were collected for the evaluation of eosinophils. Exposure to repeated low doses of allergen induced increased bronchial methacholine responsiveness 6 h after the final allergen exposure (p=0.018), and an increase in the number of eosinophils in sputum. By contrast, the late asthmatic response after challenge with a high dose of allergen was significantly attenuated by approximately 30% at 24 h after the final low-dose allergen exposure (p = 0.03). In summary, repeated low doses of allergen given directly to the airways, attenuate the high-dose allergen-induced late response, despite enhanced bronchial hyperresponsiveness to methacholine and elevated sputum eosinophils prior to allergen challenge. PMID- 11488320 TI - Is the increase in childhood asthma coming to an end? Findings from three surveys of schoolchildren in Rome, Italy. AB - Time trends in the prevalence of asthma, family history of asthma and atopy in Roman schoolchildren were assessed. The study population consisted of all children (aged 6-14 yrs) attending two primary schools in Rome, situated in urban areas that differed markedly in socioeconomic conditions and environmental pollution. Three questionnaire-based surveys were conducted in 1974, 1992 and 1998 in 2,259, 1,229 and 1,139 children. The prevalence of asthma in males and females increased significantly during 1974-1992 and remained stable from 1992 1998. In age groups born in the subsequent 4-yr periods it increased almost linearly, for children born from 1962-1965 to 1982-1985 (4.4%-12.5%), and remained remarkably stable in children born after 1985. Because the prevalence of asthma had a steeper trend in males than in females (approximately 0.55% x yr(-1) versus 0.25% x yr(-1)), the male:female asthma ratio increased (1:38 in 1974; 1:84 in 1992 and 1:62 in 1998). No single environmental factor, including area of residence, seemed to influence the prevalence of asthma. Family history of asthma and atopy also increased steadily (0.72% x yr(-1) and 0.30% x yr(-1) respectively) more than doubling during the 24-yr study period. The strong relationship between asthma and a family history of atopy not only persisted but also strengthened over time (23.3% of asthmatic children belonged to families with atopic illnesses in 1974 but 44.2% in 1998). The environmental factors that might explain the almost three-fold rise in childhood asthma between 1974 and 1992 remain unknown but the genetic background of the disease has presumably remained unchanged since the early 1970s. The fact that the prevalence of asthma increased no further during the past 6 yrs suggests that the progressive induction of asthma symptoms in genetically predisposed subjects is a self limiting process that has probably come to an end in the authors' study area. PMID- 11488321 TI - The feasibility of airways hyperresponsiveness as an inclusion criterion for studies on childhood asthma. AB - The feasibility of moderately severe airway hyperresponsiveness (AH) was examined as an inclusion criterion for clinical trials in asthmatic children. During the baseline period of a long-term clinical trial in asthmatic children, maintenance therapy with fluticasone (200 microg x day(-1)) was stopped for a maximum of 8 weeks and methacholine challenges were performed at 2-week intervals or earlier if the patients' condition deteriorated. Patients were eligible to continue the study if the provocative dose of methacholine causing a 20% fall in forced expired volume in one second (FEV1) (PD20) was <80 microg. Fifty-one per cent of the children did not develop a PD20 < 80 microg after withdrawal of fluticasone. Patients with or without a PD20 <80 microg did not differ in duration of asthma, duration of treatment, or peak flow variation. Patients with a PD20 <80 microg had higher levels of total and specific immunoglobulin-E, and lower levels of FEV1 and mean maximal expiratory flow than patients with a PD20 > or = 80 microg. Forty-four per cent of the patients with a PD20 > or = 80 microg did not have any symptoms during the wash-out period and 39% of these patients remained free from symptoms during one year follow-up. The results of this study suggest that recruiting asthmatic children for clinical trials may be difficult if airways hyperresponsiveness is used as the sole inclusion criterion. PMID- 11488322 TI - Respiratory illness and healthcare utilization in children: the primary and secondary care interface. AB - The aim of the present study was to quantify the healthcare utilization of a child population according to level of respiratory illness. A stratified random sample of 713 children was selected from respondents to a postal respiratory questionnaire, carried out in two general practice populations in 1993. Children were stratified into four groups according to the number of positive responses to five key questions. These groups were used as indicators of likelihood of asthma diagnosis. A search was made of these childrens' practice records covering a 2-yr period, to include both primary and secondary healthcare. There was a significant increase across positive response groups in the proportion of children having primary and secondary care based consultations, particularly for respiratory conditions (p = 0.001). There was also a significant increase in prescribing. Of those children considered to be "likely asthmatics" from their questionnaire responses, 8.1% (n = 31) did not receive any primary or secondary care for a respiratory problem over the 2-yr period. As the likelihood of respiratory illness increased in this population, more demand was made upon resources for the treatment of respiratory illness. Quantification of this demand enables evidence based resource allocation decisions to be made. This method of quantification could be applied in other populations. PMID- 11488323 TI - Off-line sampling of exhaled air for nitric oxide measurement in children: methodological aspects. AB - Measurement of nitric oxide in exhaled air is a noninvasive method to assess airway inflammation in asthma. This study was undertaken to establish the reference range of exhaled NO in healthy school-aged children and to determine the influence of ambient NO, noseclip and breath-holding on exhaled NO, using an off-line balloon sampling method. All children attending a primary school (age range 8-13 yrs) underwent NO measurements on two occasions with high and low ambient NO. Each time, the children performed four expiratory manoeuvres into NO impermeable balloons, with and without 10 s of breath-holding and with and without wearing a noseclip. Exhalation flow and pressure were not controlled. NO was measured within 4 h after collection, by means of chemiluminescence. All children completed a questionnaire on respiratory and allergic disorders, and performed flow/volume spirometry. With low ambient NO, the mean exhaled NO value of 72 healthy children with negative questionnaires and normal lung function was 5.1 +/- 0.2 parts per billion (ppb) versus a mean of 6.8 +/- 0.3 ppb in the remaining 49 children with positive questionnaires for asthma and allergy, and/or recent symptoms of cold (p=0.001). Exhaled and ambient NO were significantly related, especially with ambient NO > 10 ppb (r = 0.86, p=0.0001 versus r=0.34, p=0.004 for ambient values <10 ppb). The use of a noseclip, with low ambient NO and without breath-holding, caused a small decrease in exhaled NO values (p=0.001). The effect of breath-holding on exhaled NO depended on ambient NO. With ambient NO > 10 ppb, exhaled NO decreased, whereas with ambient NO < 10 ppb, exhaled NO increased after 10 s breath-hold. It is concluded that off-line sampling in balloons is a simple and, hence, attractive method for exhaled nitric oxide measurements in children which differentiates between groups with and without self-reported asthma, allergy and colds, when ambient nitric oxide is < 10 parts per billion. Wearing a noseclip and breath-holding affected measured values and should, therefore be standardized or, preferably, avoided. PMID- 11488324 TI - Natural history of sensitization, symptoms and occupational diseases in apprentices exposed to laboratory animals. AB - The natural history of the development of sensitization and disease due to high molecular-weight allergens is not well characterized. This study describes the time-course of the incidence of work-related symptoms, skin reactivity and occupational rhinoconjunctivitis (RC) and asthma (OA); and assesses the predictive value of skin testing and RC symptoms in apprentices exposed to laboratory animals, in a 3-4-yr programme. Four-hundred and seventeen apprentices at five institutions were assessed prospectively with questionnaire, skin-testing with animal-derived allergens, spirometry and airway responsiveness (n=373). Depending on the school, students were seen 8 (n=136), 20 (n=345), 32 (n=355) and 44 (n=98) months after starting the programme. At all visits, the incidence was greater for work-related RC symptoms followed in order by skin reactivity, occupational RC, and, almost equally, OA and work-related respiratory symptoms. The incidence-density figures were comparable for each follow-up period and for most indices up to 32 months after entry into the study and then tended to decrease. The positive predictive values (PPVs) of skin reactivity to work related allergens for the development of work-related RC and respiratory symptoms were 30% and 9.0%, respectively, while the PPVs of work-related RC for the development of OA was 11.4%. Sensitization, symptoms and diseases occur maximally in the first 2-3 yrs after starting exposure to laboratory animals. Skin reactivity to work-related allergens and rhinoconjuctivitis symptoms have low positive predictive values. PMID- 11488325 TI - Diesel exhaust enhances airway responsiveness in asthmatic subjects. AB - Particulate matter (PM) pollution has been associated with negative health effects, including exacerbations of asthma following exposure to PM peaks. The aim of the present study was to investigate the effects of short-term exposure to diesel exhaust (DE) in asthmatics, by specifically addressing the effects on airway hyperresponsiveness, lung function and airway inflammation. Fourteen nonsmoking, atopic asthmatics with stable disease, on continuous treatment with inhaled corticosteroids, were included. All were hyperresponsive to methacholine. Each subject was exposed to DE (particles with a 50% cut-off aerodynamic diameter of 10 microm (PM10) 300 microg x m(-3)) and air during 1 h on two separate occasions. Lung function was measured before and immediately after the exposures. Sputum induction was performed 6 h, and methacholine inhalation test 24 h, after each exposure. Exposure to DE was associated with a significant increase in the degree of hyperresponsiveness, as compared to after air, of 0.97 doubling concentrations at 24 h after exposure (p < 0.001). DE also induced a significant increase in airway resistance (p=0.004) and in sputum levels of interleukin (IL) 6 (p=0.048). No changes were detected in sputum levels of methyl-histamine, eosinophil cationic protein, myeloperoxidase and IL-8. This study indicated that short-term exposure to diesel exhaust, equal to high ambient levels of particulate matter, is associated with adverse effects in asthmatic airways, even in the presence of inhaled corticosteroid therapy. The increase in airway responsiveness may provide an important link to epidemiological findings of exacerbations of asthma following exposure to particulate matter. PMID- 11488326 TI - Observer variation in computed tomography of pleural lesions in subjects exposed to indoor asbestos. AB - To assess the reliability of computed tomography (CT) in detecting discrete pleural lesions, the interobserver and intra-observer variability in reading the conventional and high-resolution CT (HRCT) scans of 100 volunteers, who had worked for > or = 10 yrs in a building with known asbestos contamination, was evaluated. In the first session, pleural abnormalities were detected by a single radiologist (A1) in 13 subjects. In the second session, the scans were read again independently by the same radiologist (A2) and two other experienced radiologists (B, C). The final decision for the presence of pleural lesions was made in a final consensus reading. This gave a diagnosis of pleural abnormalities in 18 subjects, of whom eight (44%) had been detected by all three readers, five (28%) by two readers and four (22%) by only one reader; one scan, rated normal by all readers during the second session, was reconsidered because pleural abnormalities had been noted at the first reading (A1). The intra-observer agreement for reader A was good (kappa (kappa) 0.68) but the interobserver agreement between the readers was only fair to moderate (weighted kappa: A2-B=0.43, A2-C = 0.45, B-C = 0.26) in the second reading session. In conclusion, when looking for the prevalence of pleural lesions in indoor asbestos exposed subjects, the potential lack of consistency in reporting the presence of small pleural abnormalities must be borne in mind and strict precautions must be taken. PMID- 11488327 TI - The role of environmental and occupational exposures in Turkish immigrants with fibre-related disease. AB - Environmental exposure to tremolite and erionite causes endemic diseases of the lung and pleura in Turkey. This study aimed to evaluate the impact of these exposures and further occupational exposures on fibre-related diseases in Turkish immigrants living in Belgium. The study included 51 males and 17 females that emigrated < 1-38 yrs ago. Most of them (n=46) had nonmalignant pleural lesions, one had asbestosis and one had mesothelioma. Environmental asbestos exposure was likely for the majority of patients (60%), but there were also reports of possible occupational asbestos (n = 14) and erionite (n = 2) exposure. Tremolite was the main fibre type in bronchoalveolar lavage fluid (BALF). Elevated concentrations of amosite or crocidolite were detected in only two patients. The delay elapsed since the end of the environmental exposure had no influence on the asbestos body or the tremolite fibre concentrations in the BALF of Turkish immigrants. Most fibre-related diseases in Turkish immigrants are probably due to environmental rather than occupational exposure. Precise information about geographical origin and occupation should be obtained when investigating these patients. Mineralogical analysis of bronchoalveolar lavage fluid gives valuable information on the type and intensity of exposure, especially in patients with both environmental and occupational exposure. PMID- 11488328 TI - Factors associated with relapse after ambulatory treatment of acute exacerbations of chronic bronchitis. DAFNE Study Group. AB - This study aimed to identify the risk factors for relapse after ambulatory treatment of acute exacerbations of chronic bronchitis (AECB) that can easily be used in a primary care setting. Data were prospectively collected on 2,414 ambulatory patients with AECB from 268 general practices located throughout Spain. A multivariate model to identify risk factors independently associated with failures was developed and validated from the information recorded at the inclusion visit and at 30-days follow-up visit. A total of 507 patients relapsed (21%); of these, 84 required admission (16.5%). The multivariate model for prediction of the risk of relapse included 2,414 cases: 1,689 for the developmental sample and 725 in the validation sample. The model obtained contained three readily-obtainable variables: ischaemic heart disease (odds ratio (OR)=1.63; 95% confidence interval (CI)=1.07-2.47), degree of dyspnoea (OR = 1.31; 1.14-1.50) and number of visits to the general practitioner the previous year (OR = 1.07; 1.04-1.10). The model calibrated well in developmental and validation samples (goodness-of-fit tests: p = 0.295 and p = 0.637, respectively). Severity of the exacerbation was not associated with increased risk of relapse in either univariate or multivariate analysis. The present results suggest that baseline characteristics of the patients such as degree of dyspnoea, coexisting ischaemic heart disease and number of previous visits to the general practitioner for respiratory problems are strongly associated with increased risk of relapse after ambulatory treatment of acute exacerbations of chronic bronchitis. In contrast, exacerbation severity was not associated with clinical failure. Guidelines for management of acute exacerbations of chronic bronchitis should consider such risk factors and advocate intensive broad spectrum treatment and closer follow-up of patients exhibiting them. PMID- 11488329 TI - Exhaled nitric oxide in chronic obstructive pulmonary disease: relationship to pulmonary function. AB - The following study was undertaken in order to determine how exhaled nitric oxide (eNO) levels in former smokers with chronic obstructive pulmonary disease (COPD) compared to eNO levels in patients with asthma and in healthy nonsmoking volunteers. The study also aimed to determine any relationship between eNO levels in COPD and: 1) conventional measures of lung function; and 2) inhaled corticosteroid (ICS) use. In former smokers with COPD, nonsmokers with asthma and volunteers, eNO levels, spirometry, lung volumes, carbon monoxide diffusion capacity of the lung (DL,CO) and resting oxygen saturation (Sa,O2) were measured. Median eNO was significantly higher among patients with COPD than among healthy volunteers (p = 0.003) but lower than among patients with asthma (p < 0.01). There was no significant difference in eNO levels between COPD patients using ICS and those not using ICS. By contrast, eNO was lower among asthma patients who used ICS (median 32 parts per billion (ppb); 25-75% range 16-54) than among asthma patients who did not (51 ppb; 32-87) (p = 0.034). Among patients with COPD, eNO was inversely correlated with forced expiratory volume in one second, DL,CO and Sa,O2, and was positively correlated with the residual lung volume/total lung capacity ratio. Among patients with asthma, no significant correlations were found. Exhaled nitric oxide is increased in patients with chronic obstructive pulmonary disease, an increase that is influenced by structural abnormalities of tobacco-induced lung damage. PMID- 11488330 TI - Metabolic characteristics of the deltoid muscle in patients with chronic obstructive pulmonary disease. AB - The purpose of this study was to analyse key enzyme activities of the deltoid muscle (DM) in chronic obstructive pulmonary disease (COPD) patients. The activities of one oxidative enzyme (citrate synthase (CS)), two glycolytic enzymes (lacatate dehydrogenase (LD); and phosphofructokinase (PFK)) and one enzyme related to the use of energy stores (creatine kinase (CK)) were determined in the DM of 10 patients with COPD and nine controls. Exercise capacity (cycloergometry) and the handgrip strength were also evaluated. Although exercise capacity was markedly reduced in COPD (57 +/- 20% predicted), their handgrip strength was relatively preserved (77 +/- 19% pred). The activity of LD was higher in the COPD patients (263.9 +/- 68.2 versus 184.4 +/- 46.5 mmol x min(-1) x g(-1), p<0.01), with a similar trend for CS (67.3 +/- 33.3 versus 46.0 +/- 17.4 mmol x min(-1) x g(-1), p = 0.07). Interestingly, the activity of the latter enzyme was significantly higher than controls if only severe COPD patients were considered (81.8 +/- 31.2 mmol x min(-1) x g(-1), p < 0.01). PFK and CK activities were similar for controls and COPD. Chronic obstructive patients show a preserved or even increased (severe disease) oxidative capacity in their deltoid muscle. This coexists with a greater capacity in the anaerobic part of the glycolysis. These findings are different to those previously observed in muscles of the lower limbs. PMID- 11488331 TI - Lymphocyte population and apoptosis in the lungs of smokers and their relation to emphysema. AB - Previously, it had been shown that T-lymphocytes are the predominant inflammatory cells found in the alveolar wall of smokers and their numbers correlated with the extent of emphysema. However, the phenotype of these cells was not defined. The aim of this study was to describe the different T-cell phenotypes and investigate the possible presence of apoptosis in the lung parenchyma of smokers. Samples from lungs were obtained at surgery from 15 patients who smoked and six who had never smoked. Samples were frozen and prepared for histological and immunocytochemical examination. Slides were stained for CD3+, CD4+, CD8+, gammadelta T-cells, CD56 natural killers ((NK) cells), and elastase (neutrophils). Anti-CD95 monoclonal antibodies and in situ end-labelling techniques were used to detect Fas expression and apoptosis. Positive staining cells were expressed as cells-mm alveolar wall-, percentage of total cells, and Fas/APO and apoptosis index. Emphysema was identified macroscopically, microscopically and reported as present or absent. All subjects had pulmonary function tests before surgery. Neutrophils were the predominant cell in the lung parenchyma of nonsmokers and smokers without emphysema. In smokers with emphysema, the CD3+ and CD8+ were the predominant cells (p<0.05) in the alveolar wall. gammadelta cells were increased in all smokers and no increased numbers of NK cells was found. The T-cell numbers x mm alveolar wall(-1) showed a bilinear relationship with the amount smoked increasing at an inflection point of 30 packs yr(-1) (R2= 0.345; p < 0.01). Apoptosis in smokers showed a bilinear relationship with the amount smoked increasing sharply in smokers with emphysema (R2=0.3613; p < 0.009). It is concluded that the pathogenesis of emphysema might be mediated by T-lymphocytes, mainly CD8+ cytolytic T-cells, and that apoptosis might be one of the mechanisms of lung destruction leading to the development of emphysema. If this is the case, it could be speculated that T-cell inflammation is a response to antigenic stimuli originating in the lung and induced by cigarette smoking. PMID- 11488332 TI - Quality of life of idiopathic pulmonary fibrosis patients. AB - Little attention has been paid to quality of life (QOL) in idiopathic pulmonary fibrosis (IPF). Therefore, the aim of this study was to address this issue and study the relationship between QOL, depressive symptoms, and breathlessness in these patients. Forty-one IPF patients and 41 healthy persons matched for age and sex completed the World Health Organization Quality of Life assessment instrument 100. The IPF patients also completed the Beck Depression Inventory, the Bath Breathlessness Scale, a social support questionnaire and a question concerning perceived seriousness of illness. Compared to the control group, QOL in IPF patients was mainly impaired in the domains "physical health" and "level of independence". A number of relationships were found between pulmonary function tests and QOL. The QOL facet "negative feelings" was highly associated with scores on depression. Subjective breathlessness was related to depressive symptoms and QOL. Moreover, sex and effective/emotional breathlessness predicted overall QOL. In conclusion, the impairment of the quality of life areas "physical health" and "level of independence" are important issues in idiopathic pulmonary fibrosis. Subjective breathlessness, especially the effective/emotional scale, seems related to quality of life and depressive symptoms. Rehabilitation programmes are needed that are aimed at physiological aspects and psychosocial aspects of idiopathic pulmonary fibrosis in order to enhance the quality of life of these patients. PMID- 11488333 TI - Eotaxin and monocyte chemoattractant protein-1 in chronic eosinophilic pneumonia. AB - Chronic eosinophilic pneumonia (CEP) is characterized by chronic or recurrent pulmonary infiltrates with eosinophils, but the precise mechanism of eosinophil accumulation has not been fully elucidated. Eotaxin is one of the CC chemokines that selectively recruits eosinophils and contributes to the pathogenesis of allergic airway diseases including asthma, but its roles in pathogenesis of CEP have not been fully elucidated. The authors measured concentrations of eotaxin and other CC chemokines, monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted, macrophage inflammatory protein 1alpha, and the eosinophil activating Th2 cytokine interleukin (IL)-5 in bronchoalveolar lavage (BAL) fluid from CEP patients (n=11), and compared these concentrations with those from control subjects (n = 6). The eotaxin (904 +/- 203 versus 29 +/- 7 pg x mL(-1), p = 0.0001), MCP-1 (194 +/- 57 versus 15 +/- 2 pg x mL(-1), p < 0.05), and IL-5 (7.8 +/- 2.0 versus 2.7 +/- 0.6 pg x mL(-1), p < 0.05) levels were significantly higher for cases with CEP in comparison to those serving as controls. Proportions of eosinophil and lymphocyte counts were greater in BAL fluid from CEP patients. Eotaxin and IL-5 levels correlated with the proportion of eosinophils in BAL fluid from CEP patients. MCP-1 correlated with the relative lymphocyte numbers. In short, eotaxin, interleukin-5, and monocyte chemoattractant protein-1 levels were higher in the BAL fluid of CEP patients and these levels may contribute to eosinophil and lymphocyte recruitment and activation in the airways as found with this disorder. PMID- 11488334 TI - Expression of discoidin domain receptor 1 tyrosine kinase on the human bronchial epithelium. AB - Discoidin domain receptor 1 (DDR1) tyrosine kinases constitute a novel family of receptors characterized by a unique structure in the ectodomain (discoidin-I domain). The DDR1 ligand is the extracellular matrix protein collagen. To identify receptor tyrosine kinases (RTKs) involved in control of growth and differentiation of human bronchial epithelial (HBE) cells, a polymerase chain reaction-based search for RTKs in HBE cells was performed. DDR1 was the most abundant clone identified. Northern analysis detected a 3.6 kb DDR1 messenger ribonucleic acid (mRNA) expressed in HBE cells and transformed HBE lines, BET-1A and BEAS-2B. In addition, fluorescence-activated cell sorter (FACS) analyses using an anti-DDR1 antibody showed that DDR1 was expressed on HBE cells and two HBE lines. Immunohistochemical staining using human bronchial tissue demonstrated that DDR1 was mainly expressed at the basolateral cell surface of the bronchial epithelium. Furthermore, immunostaining of type IV collagen, a major component of the basement membrane, clearly showed that the basement membrane was closely attached to the basal surface of the bronchial epithelium. Since collagen binds to and activates discoidin domain receptor 1 tyrosine kinase, colocalization of discoidin domain receptor 1 and its ligand type IV collagen demonstrates a potential interaction of discoidin domain receptor 1 on the bronchial epithelium with type IV collagen. Further study of this interaction may define the functional significance of the collagen-discoidin domain receptor 1 signalling pathway in health and in disease. PMID- 11488335 TI - Endothelin-1-induced airway and parenchymal mechanical responses in guinea-pigs: the roles of ETA and ETB receptors. AB - Endothelin-1 (ET-1) has been shown to have a constrictor effect on the airways and parenchyma; however, the roles of the ETA and ETB receptors in the ET-1 induced changes in the airway and tissue compartments have not been fully explored. Low-frequency pulmonary impedance (ZL) was measured in anaesthetized, paralysed, open-chest guinea-pigs. ZL spectra were fitted by a model to estimate airway resistance (Raw) and inertance (Iaw), and coefficients of tissue damping (G) and elastance (H), and hysteresivity (eta = G/H). Two successive doses of ET 1 (0.05 and 0.2 nmol x kg(-1)) each evoked significant dose-related increases in Raw, G, H and eta. Pretreatment with 20 nmol x kg(-1) BQ-610 (a highly selective ETA receptor antagonist) resulted in a significantly decreased elevation only in H after the lower dose of ET-1. However, all parameters changed significantly less on the administration of ET-1 after pretreatment with 80 nmol-kg(-1) BQ-610, with 20 nmol x kg(-1) ETR-P1/fl (a novel ETA receptor antagonist) or with 20 nmol x kg(-1) IRL 1038 (an ETB receptor antagonist). The results of the separate assessments of the airway and tissue mechanics demonstrate that endothelin-1 induces airway and parenchymal constriction via stimulation of both receptor types in both compartments. PMID- 11488336 TI - Epidemiology of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a leading cause of world-wide mortality and disability. On average approximately 5-15% of adults in industrialized countries have COPD defined by spirometry. In 1990, COPD was considered to be at the twelfth position world-wide as a cause of combined mortality and disability but is expected to become the fifth cause by the year 2020. COPD has a chronic long-lasting course characterized by irreversible decline of forced expiratory volume in one second (FEV1), increasing presence of dyspnoea and other respiratory symptoms, and progressive deterioration of health status. After diagnosis the 10-yr survival rate is approximately 50% with more than one-third of patients dying due to respiratory insufficiency. Several environmental exposures such as air pollution increase the risk of death in COPD patients. The aetiology of COPD is overwhelmingly dominated by smoking although many other factors could play a role. Particular genetic variants are likely to increase the susceptibility to environmental factors although little is known about which are the relevant genes. There is clear evidence about the role of the alpha-1-antitrypsin but the fraction of COPD attributable to the relevant variants is only 1%. Phenotypic traits that are considered to play a role in the development of COPD include sex, with females being at a higher risk, bronchial responsiveness and atopy. There is strong causal evidence regarding the relationship between smoking and COPD with decline in FEVI levelling off after smoking cessation. Passive smoking has been found to be associated with a small though statistically significant decline in FEV1. Other risk factors that are likely to be relevant in the development of COPD are occupation, low socioeconomic status, diet and possibly some environmental exposures in early life. Although there is accumulating evidence that oxygen therapy, pharmacological treatment and rehabilitation may improve the course of chronic obstructive pulmonary disease, preventing smoking continues to be the most relevant measure, not only to prevent chronic obstructive pulmonary disease, but also to arrest its development. PMID- 11488337 TI - Bacteria, antibiotics and COPD. AB - Bacterial infection is one of several important causes of exacerbations of chronic obstructive pulmonary disease (COPD) that may coexist. COPD is a heterogeneous condition and the incidence of bacterial infection is not uniform; mucus hypersecretion may be an important risk factor. The bacteriology of infections varies depending on the severity of the underlying airway disease. There is now a much better understanding of the pathogenesis of bacterial infections of the respiratory mucosa. Lower airway bacterial colonization may be a stimulus for chronic inflammation and may influence the interval between exacerbations. Antibiotic resistance has increased in all the major pathogens. Antibiotics are an important part of the treatment of acute exacerbations of COPD and the decision about whether to give an antibiotic can be made on clinical grounds. It is more difficult to decide, on the available evidence, whether patient characteristics and the risk of antibiotic resistance should influence choice of empiric antibiotic treatment. Most new antibiotics are modifications of existing structures, suggesting that every effort should be made to conserve the sensitivity of current antibiotics by using them appropriately. PMID- 11488338 TI - FMD: DEFRA offers advice on restructuring and restocking. PMID- 11488339 TI - Self-regulation and the veterinary profession. PMID- 11488340 TI - Prevalence and public health significance of blemishes in cuts of Irish beef. AB - Between February 1999 and February 2000 more than 45,000 quarters of Irish beef cattle were examined for the presence of blemishes as they were cut into primal and subprimal cuts. A total of 2,379 blemishes, either scars or cysts, were found. Eight of the scars and one cyst tested positive for residues of inhibiting substances. PMID- 11488341 TI - Ultrasonographic examination of the caecum and the proximal and spiral ansa of the colon of cattle. AB - The large intestine of 10 cows was examined from the right abdominal wall with a 3.5 MHz linear transducer. The cows were then slaughtered, and the organs were re examined in a water bath. The caecum was visualised from the middle region of the abdominal wall. It ran caudo-cranially, varied in diameter from 5.2 to 18.0 cm and was situated immediately adjacent to the abdominal wall. The lateral wall of the caecum appeared as a thick, echogenic, crescent-shaped line. It could be visualised as far cranially as the 12th intercostal space. Although its junction could not be identified, the proximal ansa of the colon was recognised on the basis of its anatomical position and its diameter, which was smaller than that of the caecum. The spiral ansa of the colon and the descending colon were situated dorsal to the caecum and could be identified by moving the transducer horizontally along the abdominal wall to the last rib. The spiral ansa of the colon was situated ventral to the descending colon, and its walls appeared as thick echogenic lines. In a contracted state, the spiral colon had the appearance of a garland. PMID- 11488342 TI - Progressive encephalomyelopathy and cerebellar degeneration in 10 captive-bred cheetahs. AB - Progressive ataxia, with head tremor, developed in 10 captive-born cheetah cubs under six months of age. The condition was usually preceded by coryza and an ocular discharge. Initially the ataxia and weakness affected the hindquarters, then the forelegs, and head tremor developed later. Significant pathological changes were confined to the central nervous system. There was widespread Wallerian degeneration in the funiculi of the spinal cord (except those in the dorsal columns), in the medulla and in the cerebellum. In the cerebellum there was degeneration of Purkinje cells and of the molecular and granular cell layers. There was chromatolysis in the Purkinje cells, the ventral horn cells of the spinal cord and in the neurons of the lateral vestibular nucleus. The olivary nucleus was necrotic. There were foci of inflammatory cells in the molecular layer of the cerebellum and in the medulla. The cause of the disease remains unknown. PMID- 11488343 TI - Prevalence of BSE in western France by screening cattle at risk: preliminary results of a pilot study. PMID- 11488344 TI - Serological evidence of avian pneumovirus infection in reared and free-living pheasants. PMID- 11488345 TI - Avermectin resistance in Cooperia pectinata in cattle in Argentina. PMID- 11488346 TI - Prenatal infection with a hydatid cyst in a camel (Camelus dromedarius). PMID- 11488347 TI - Kidney transplants in cats. PMID- 11488348 TI - Kidney transplants in cats. PMID- 11488349 TI - Kidney transplants in cats. PMID- 11488350 TI - Veterinary education and training for 2010 and beyond. PMID- 11488351 TI - vCJD in Leicestershire. PMID- 11488352 TI - Avermectin resistance of the cattle tick Boophilus microplus in Brazil. PMID- 11488353 TI - Normalization to lithium for the assessment of metal contamination in coastal sediment cores from the Aegean Sea, Greece. AB - Sediment cores from the harbour and the coastal zone of Mytilene, island of Lesvos, Greece, were used to study the metal contamination caused by the discharge of untreated urban effluents into the sea. In the harbour. the upper layers were highly enriched in Cd, Cu, Pb and Zn, while no metal enrichment was recorded in the cores from the wider coastal zone. The metal data were normalized to Li (conservative element) to compensate for the natural textural and mineralogical variability. It was found that only the upper 18 cm of the core collected from the harbour of Mytilene could be reported as metal contaminated. Also, through the normalization procedure, it was found that the surface layers of coastal sediments assumed 'clean' were enriched in Pb, probably as a result of atmospheric transportation of the metal from the nearby town. PMID- 11488354 TI - Seasonal variation of antioxidant and biotransformation enzymes in barnacle, Balanus balanoides, and their relation with polyaromatic hydrocarbons. AB - Seasonal variations in the antioxidant enzymes (catalase, superoxide dismutase [SOD], NADH-DT diaphorase), biotransformation enzyme, glutathione-S-transferase (GST) and microsomal lipid peroxidation in digestive tissue of barnacle, Balanus balanoides, from polluted and non-polluted populations have been evaluated. Relationships with accumulated polyaromatic hydrocarbon (PAH) concentration in barnacle tissues and environmental parameters (water temperature, salinity, dissolved oxygen concentration, water pH) were determined. As a general trend, maximum antioxidant enzyme and GST activities were detected in the pre-monsoon period or summer (March-June) followed by a gradual decrease during the monsoon (July October) with a minimum in the post-monsoon period or winter (November February). This pattern was similar to tissue concentrations of PAHs, resulting in a significant positive correlation with antioxidant enzymes, mainly catalase and SOD. Microsomal lipid peroxidation exhibited an almost reverse trend of seasonal variation to that of antioxidant enzyme activities indicating an enhanced susceptibility of barnacle tissues to oxidative stress. Among the environmental parameters, only water temperature seemed to have a significant effect on observed variations of antioxidant enzymes and GST activities. The barnacles from polluted and non-polluted populations exhibited seasonal differences in the activities of all the enzymes studied, particularly catalase, SOD and GST, suggesting the possibility of some biochemical adaptation in organisms from a chronically polluted environment. The results indicated that antioxidant defense components, catalase and SOD, are sensitive parameters that could be useful biomarkers for the evaluation of contaminated aquatic ecosystems. The results also suggested the potentiality of barnacle, B. balanoides, as a bioindicator organism against organic pollution. PMID- 11488355 TI - Macrofauna impact on Ulva rigida C. Ag. production and relationship with environmental variables in the lagoon of Venice. AB - The grazing pressure of the macrofaunal invertebrates associated with the biomass of Ulva rigida in the Venice lagoon, their species composition and relationship with environmental variables have been studied. Fifteen sampling campaigns were carried out during different seasons both in the central basin of the lagoon, dominated by macroalgae (especially U. rigida C. Ag.) and in the southern basin, prevalently populated by seagrasses (especially Zostera marina L.). Replicate experiments were conducted in the field by exposing Ulva fronds in net cages of 10 and 1 mm (control) mesh-sizes to allow or prevent grazer entrance. The grazing pressure was determined as Ulva growth rate difference in the cages. In the absence of invertebrate herbivores, Ulva exhibited per cent relative growth rates (%RGRs) ranging from 1.5 to 9.5% day(-1), whereas in their presence the %RGRs were significantly lower (from -2.5 to 3.4% day(-1)) and frequently negative, especially in the station dominated by macroalgae. In this area, peak grazing rates and macrofauna biomasses of up to 8.6% day(-1) and 1,480 g m(-2) fwt (84.4 ash-free dry weight), respectively, were found. On the whole, during in field experiments in the Ulva-dominated station, herbivores removed an amount of biomass whose percentage ranged from 59 to 165% (mean: 103%) of the biomass yield (grazers excluded) found in the cages. These results suggest the possibility that grazers could act as an important factor affecting Ulva production in the Venice lagoon. Macrofauna populations were analysed by means of multivaliate techniques applied to biological variables only and biological and environmental variables together. Data of individual abundance, after a log(x + 1) transformation and the calculation of the Bray-Curtis matrix, were classified using the Cluster Analysis and ordinated by means of the Non-Metric Multidimensional Scaling (MDS) technique, in accordance with the strategies used in the study of multispecies distributions. Finally, biological and environmental variables were analysed together by means of correlation matrices and the Principal Component Analysis. PMID- 11488356 TI - Historical contamination of PAHs, PCBs, DDTs, and heavy metals in Mississippi River Delta, Galveston Bay and Tampa Bay sediment cores. AB - Profiles of trace contaminant concentrations in sediment columns can be a natural archive from which pollutant inputs into coastal areas can be reconstructed. Reconstruction of historical inputs of anthropogenic chemicals is important for improving management strategies and evaluating the success of recent pollution controls measures. Here we report a reconstruction of historical contamination into three coastal sites along the US Gulf Coast: Mississippi River Delta, Galveston Bay and Tampa Bay. Within the watersheds of these areas are extensive agricultural lands as well as more than 50% of the chemical and refinery capacity of the USA. Despite this pollution potential, relatively low concentrations of trace metals and trace organic contaminants were found in one core from each of the three sites. Concentrations and fluxes of most trace metals found in surface sediments at these three sites, when normalized to Al, are typical for uncontaminated Gulf Coast sediments. Hydrophobic trace organic contaminants that are anthropogenic (polycyclic aromatic hydrocarbons, DDTs, and polychlorinated biphenyls) are found in sediments from all locations. The presence in surface sediments from the Mississippi River Delta of low level trace contaminants such as DDTs, which were banned in the early 1970's, indicate that they are still washed out from cultivated soils. It appears that the DDTs profile in that sediment core was produced by a combination of erosion processes of riverine and other sedimentary deposits during floods. Most of the pollutant profiles indicate that present-day conditions have improved from the more contaminated conditions in the 1950-1970's, before the advent of the Clean Water Act. PMID- 11488357 TI - Expression of P-glycoprotein in southeastern oysters, Crassostrea virginica. AB - These studies provide important fundamental information regarding the expression of P-glycoprotein (p-gp) in southeastern oysters (Crassostrea virginica). Using rhodamine transport studies, p-gp activity was detected in newly fertilized embryos. A monoclonal antibody (C219) was used to evaluate p-gp expression in oyster tissues. On the basis of laboratory studies, p-gp expression tended to be higher in gill tissues than mantle tissues, and was generally not related to salinity differences. Seasonal studies were conducted with oysters collected monthly for 1 year from Lighthouse Creek, an unpolluted site. There was a general pattern of higher p-gp expression in the warmer months and lower expression in the colder months. In contrast, total gill protein concentrations decreased during the warmer months and increased during the colder months. These studies indicate that there are seasonal patterns in p-gp expression which may represent an adaptive response to natural stressors associated with summer conditions. PMID- 11488358 TI - Fibromyalgia: the enigma and the stigma. PMID- 11488359 TI - Bupropion sustained release as a smoking cessation treatment in remitted depressed patients maintained on treatment with selective serotonin reuptake inhibitor antidepressants. AB - BACKGROUND: Patients with depressive disorders smoke tobacco more often than the population at large and find quitting more difficult. Furthermore, when they quit smoking, they are more likely to suffer a relapse of depression. We evaluated the addition of bupropion sustained release (SR) for smoking cessation among patients with a history of depressive disorders being maintained in a euthymic state with selective serotonin reuptake inhibitor (SSRI) antidepressants. METHOD: Twenty five adults with DSM-IV major depressive disorder or depressive disorder NOS currently receiving SSRI maintenance treatment and smoking > or = 15 cigarettes per day participated in the 9-week study. Bupropion SR, 150 mg/day, was added to SSRI treatment and increased to 300 mg/day. Subjects were counseled on smoking cessation measures and chose a target quit date 2 or 4 weeks after the initiation of bupropion SR. Self-reported smoking status, expired carbon monoxide (CO) measurements, Hamilton Rating Scales for Depression and Anxiety scores, and weight were measured at each visit. Subjects were abstinent if they reported not smoking during the prior 7 days, confirmed with an expired-air CO value of < or = 10 ppm. RESULTS: Eight (32%) of 25 subjects were abstinent after 9 weeks. At 3 month follow-up, 3 subjects remained abstinent, 3 relapsed, and 2 were lost to follow-up. Eleven subjects (44%) were nonresponders, and 6 (24%) dropped out prior to 3 weeks of treatment due to side effects (N = 3) or were lost to follow up (N = 3). Mean weight gain was approximately 0.5 lb (0.2 kg) for those completing 9 weeks of bupropion SR treatment. During the 9-week study and the 3 month follow-up, there was no evidence of emergent depression in any subject. Four subjects (16%) spontaneously reported an improvement in SSRI-associated sexual dysfunction. CONCLUSION: These open data suggest modest effectiveness for and the safety of bupropion SR as a smoking cessation agent in individuals with depression maintained on treatment with SSRIs. Minimal weight gain, lack of emergent depressive episodes, and improvement of SSRI-associated sexual dysfunction are added advantages. PMID- 11488360 TI - Olanzapine therapy in treatment-resistant psychotic mood disorders: a long-term follow-up study. AB - BACKGROUND: Recent studies suggest a role for the atypical antipsychotic olanzapine in the acute treatment of psychotic mood disorders, but long-term data are unavailable. The purpose of this naturalistic study was to determine the long term effectiveness and tolerability of olanzapine as add-on therapy in psychotic mood disorders. METHOD: Hospital records were reviewed for 125 inpatients at the state psychiatric hospital in Buffalo, N.Y., who received at least 6 weeks of add on olanzapine treatment for psychotic mood disorders (schizoaffective disorders [bipolar and depressive type], bipolar disorders [I, II, and NOS], and major depressive disorder). A group of schizophrenic patients served as a control group (N = 50). Baseline measures, including age, gender, number of hospitalizations in the 2 years prior to olanzapine treatment, concomitant medications, the Clinical Global Impressions scale (CGI), and the Global Assessment of Functioning Equivalent (GAF-EQ) and Kennedy Axis V psychological impairment, violence, social skills, and activities of daily living subscale scores, were obtained. Follow-up information was obtained from the patients at least 6 months after initiation of olanzapine or by chart review and discussion with the treating psychiatrist. Patients with a diagnosis of psychotic mood disorders were compared with patients with the non-affective psychotic disorder (schizophrenia) on a variety of outcome measures. RESULTS: Follow-up information was available on 102 patients (82%). Mean follow-up was 15 months; 50 (49%) of the 102 patients remained on olanzapine treatment at follow-up (32 psychotic mood disorder, 18 schizophrenic). The primary reason for discontinuation in both groups was lack of response. Both the psychotic mood disorder and schizophrenic groups had comparable outcomes on the CGI and GAF-EQ. Improvement on the Kennedy Axis V psychological impairment and social skills subscales was seen only in the psychotic mood disorders group (p < .01); both groups showed significant (p < .02) improvement in the violence subscale. Sustained mood-stabilizing effect was evident in only 7/27 (26%) of the psychotic mood disorders patients continuing on add-on olanzapine treatment at follow-up. CONCLUSION: Lack of response was the primary reason for discontinuation of add-on olanzapine in both groups. Mood symptoms predicted a better response to add-on olanzapine in patients with psychotic mood disorders on selective outcome measures. However, only 26% of the patients with psychotic mood disorders sustained a clinically meaningful mood-stabilizing effect with add-on olanzapine treatment at follow-up. PMID- 11488361 TI - Prevalence and clinical features of body dysmorphic disorder in adolescent and adult psychiatric inpatients. AB - BACKGROUND: The rate of body dysmorphic disorder (BDD) in inpatient psychiatric settings and the nature of the presenting complaints are unknown. Because of the shame and humiliation that BDD patients suffer, we hypothesized that, unless specifically screened for at the time of admission, BDD would be underdiagnosed in psychiatric inpatients. METHOD: 101 consecutive adult patients and 21 consecutive adolescent patients presenting for psychiatric inpatient admission to a university teaching hospital participated in the study. Subjects completed the Body Dysmorphic Disorder Questionnaire, a brief self-report measure that screens for BDD, and a follow-up interview was conducted using a reliable clinician administered semistructured diagnostic instrument for DSM-IV BDD. Data concerning current diagnoses, number of hospitalizations, number of suicide attempts, and current level of functioning were also obtained. RESULTS: Sixteen (13.1%) of the 122 subjects were diagnosed with BDD. None of the subjects with BDD had been diagnosed with BDD by their treating physician during hospitalization. All 16 subjects reported that they would not raise the issue with their physician unless specifically asked due to feelings of shame. CONCLUSION: These preliminary results suggest that BDD, an underrecognized and often severe psychiatric disorder, may be relatively common in the psychiatric inpatient setting. It is important that clinicians specifically inquire about BDD because patients will not voluntarily raise these concerns. The comorbidity of this disorder with other psychiatric illnesses may have treatment implications. PMID- 11488362 TI - Efficacy of venlafaxine extended release in patients with major depressive disorder and comorbid generalized anxiety disorder. AB - BACKGROUND: A subset of patients with comorbid major depressive disorder and generalized anxiety disorder (GAD) was examined from a double-blind. placebo controlled study comparing the efficacy and safety of venlafaxine extended release (XR) and fluoxetine. METHOD: From a total of 368 patients, 92 patients meeting DSM-IV criteria for major depressive disorder who also had comorbid GAD were identified. The comparison group comprised 276 evaluable noncomorbid patients. Patients received venlafaxine XR (75-225 mg/day), fluoxetine (20-60 mg/day), or placebo for 12 weeks. Efficacy evaluations included Hamilton Rating Scale for Depression (HAM-D), Hamilton Rating Scale for Anxiety (HAM-A), and Clinical Global Impressions (CGI) scale. RESULTS: By the final assessment at week 12, comorbid patients in the venlafaxine XR group, but not in the fluoxetine group, showed a significantly greater decrease than those in the placebo group in the primary efficacy variables of mean HAM-D and HAM-A total scores (p < .05, pairwise comparison). In comorbid patients, significant pairwise differences were noted between venlafaxine XR and placebo at week 12 for the secondary variables of HAM-D anxiety-somatization and retardation factors, HAM-D depressed mood item. HAM-A psychic anxiety factor, the Hospital Anxiety and Depression scale (HAD) anxiety subscale score, and the Covi Anxiety Scale score. Fluoxetine was significantly different from placebo only on the HAD depression subscale score. Response, defined as > or = 50% decrease in symptoms score, was achieved in 66% and 59% of the comorbid patients for HAM-D and HAM-A, respectively, in the venlafaxine XR group at week 12. This response was higher than that seen with fluoxetine (52% and 45%) or placebo (36% and 24%). Onset of efficacy appeared to be slower in comorbid than in noncomorbid patients. CONCLUSION: This is the first evidence from a controlled study of the effectiveness of pharmacotherapy in patients with comorbid major depressive disorder and GAD. The delayed improvement in comorbid patients compared with noncomorbid patients suggests that a longer treatment period may be necessary in comorbid patients. PMID- 11488363 TI - An open trial of divalproex sodium in autism spectrum disorders. AB - BACKGROUND: Autism spectrum disorders are characterized by core deficits in social interaction and speech/communication skills, repetitive behaviors, and restricted interests. Other abnormalities include seizures, electroencephalographic (EEG) abnormalities, affective instability, impulsivity, and aggression. Divalproex sodium is indicated as both an anticonvulsant in epilepsy and a mood stabilizer in bipolar illness and thus might be useful for these complicating symptoms in autism. METHOD: A retrospective pilot study was conducted to determine whether divalproex sodium was effective in treating core dimensions and associated features of autism. Fourteen patients who met DSM-IV criteria for autism, Asperger's disorder, or pervasive developmental disorder not otherwise specified, both with and without a history of seizure disorders or EEG abnormalities, were openly treated with divalproex sodium. Improvement was assessed via the Clinical Global Impressions-Improvement scale. RESULTS: Of 14 patients who completed a trial of divalproex sodium, 10 (71%) were rated as having sustained response to treatment. The mean dose of divalproex sodium was 768 mg/day (range, 125-2500 mg/day), and it was generally well tolerated. Improvement was noted in core symptoms of autism and associated features of affective instability, impulsivity, and aggression. CONCLUSION: Divalproex sodium may be beneficial to patients with autism spectrum disorders, particularly those with associated features of affective instability, impulsivity, and aggression as well as those with a history of EEG abnormalities or seizures. Of note, all patients with an abnormal EEG and/or seizure history were rated as responders. However, these findings must be interpreted with caution, given the open retrospective nature of the study. Controlled trials are needed to replicate these preliminary findings. PMID- 11488364 TI - Long-term follow-up and predictors of clinical outcome in obsessive-compulsive patients treated with serotonin reuptake inhibitors and behavioral therapy. AB - BACKGROUND: The objective of this study was to examine the long-term course of obsessive-compulsive disorder (OCD) in patients treated with serotonin reuptake inhibitors (SRIs) and behavioral therapy and to identify predictors of clinical outcome. METHOD: Sixty outpatients meeting DSM-II-R or DSM-IV criteria for OCD were followed up for 1 to 5 years (mean = 2.5 years). All of them received prolonged pharmacologic therapy with an SRI. RESULTS: Thirty-seven patients (61.7%) completed an adequate behavioral treatment. At long-term assessment, 22 patients (36.7%) exhibited a global Yale-Brown Obsessive Compulsive Scale (Y BOCS) score greater than 16 or a final reduction in Y-BOCS global score of less than 35% and were considered nonresponders. Patients who completed behavioral therapy showed a significant decrease in Y-BOCS compulsions subscale score (p = .01), whereas no significant differences in either Y-BOCS global or obsessions subscale scores between those who did and those who did not undergo behavioral therapy were detected. Obsessions of sexual/religious content were the unique factor related to a poorer long-term outcome. CONCLUSION: A substantial number of OCD patients showed persistent disabling symptoms at the long-term follow-up in spite of combined pharmacologic and behavioral treatment. Major benefits from behavioral therapy appeared to be the improvement of ritualistic behaviors. Sexual/religious obsessions predicted poorer long-term outcome, whereas short term response to SRI treatment failed to achieve predictive value in the long term course of OCD. PMID- 11488365 TI - Comparison of sexual dysfunction in male schizophrenic patients maintained on treatment with classical antipsychotics versus clozapine. AB - BACKGROUND: Antipsychotic treatment is frequently associated with sexual dysfunction. The objective of the present study was to evaluate and compare sexual function and behavior in male schizophrenic patients who regularly take either classical neuroleptic drugs or the prototypical atypical antipsychotic agent, clozapine. METHOD: Participants included 60 schizophrenic male patients (DSM-IV criteria); 30 maintained on treatment with classical antipsychotics and 30 on treatment with clozapine. The patients were evaluated with a detailed 18 item sexual function questionnaire. RESULTS: Both groups reported sexual dysfunction, although scores were significantly higher, indicating better functioning, in the clozapine-treated group in the domains of orgasmic function (number of orgasms per month, p = .037; frequency of orgasm during sex, p = .046), enjoyment of sex (p = .013), and sexual satisfaction (p = .0004). Equivocal results were obtained for the desire parameters. CONCLUSION: Maintenance therapy with the atypical neuroleptic clozapine may be associated with a lesser degree of sexual dysfunction than the classical antipsychotics in male outpatients with chronic schizophrenia. PMID- 11488366 TI - Adherence assessments and the use of depot antipsychotics in patients with schizophrenia. AB - BACKGROUND: Antipsychotic medications significantly ameliorate the symptoms of schizophrenia, but patients are often noncompliant with these medications. Research evidence supports the use of depot antipsychotics in noncompliant patients. METHOD: Between January 9, 1991, and December 19, 1995, 1307 veterans with schizophrenia or schizoaffective disorder (ICD-9) were enrolled in a study of enhanced psychosocial programming at 14 Veterans Administration Medical Centers. All had a history of high inpatient use. At enrollment, clinicians listed patient medications, rated patient compliance, and completed a Brief Psychiatric Rating Scale (BPRS) and Global Assessment of Functioning (GAF). Patients reported medication side effects. We describe depot antipsychotic use among these patients and examine the relationship between depot use, assessed compliance, and patient characteristics. RESULTS: At enrollment, 18% of patients in this cohort were receiving depot antipsychotics; however, clinicians reported that 49% had been noncompliant with medication in the past year. Depot use varied significantly with treatment site; African Americans were more likely to receive depot antipsychotics and less likely to receive atypical antipsychotics than white patients. Patients on depot and oral agents had similar levels of psychiatric symptoms, but patients on depot antipsychotics were more likely to receive high doses and complain of side effects. CONCLUSION: Clinicians prescribed depot antipsychotics relatively infrequently, despite high rates of noncompliance and high levels of inpatient use. Variation in use with treatment site and ethnic group suggests barriers to implementing research-based recommendations for depot use in noncompliant patients. Quality improvement programs should consider facilitating the appropriate use of depots. PMID- 11488367 TI - Electroconvulsive therapy in medication-nonresponsive patients with mixed mania and bipolar depression. AB - BACKGROUND: The aim of this study was to investigate the effectiveness of electroconvulsive therapy (ECT) in medication-nonresponsive patients with mixed mania and bipolar depression. METHOD: Forty-one patients with mixed mania (DSM-IV diagnosis of bipolar I disorder, most recent episode mixed) and 23 patients with bipolar depression (DSM-IV diagnosis of bipolar I disorder, most recent episode depressed) consecutively assigned to ECT treatment were included in this study. Subjects were evaluated using the Montgomery-Asberg Depression Rating Scale (MADRS), the Brief Psychiatric Rating Scale (BPRS), and the Clinical Global Impressions-Severity of Illness scale (CGI-S). Assessments were carried out the day before starting ECT, 48 hours after completion of the third session (T1), and a week after the last session of ECT (T2). RESULTS: Both groups received an equal number of ECT sessions (mean +/- SD = 7.2 +/- 1.7 vs. 7.3 +/- 1.6). In both groups, within-group comparisons showed that there was a significant reduction in CGI-S score (mixed mania, p <.0001 at T1 and T2; bipolar depression, p < .01 at T1, p < .0001 at T2), MADRS total score (both groups, p < .0001 at T1 and T2), BPRS total score (mixed mania, p < .0001 at T1 and T2; bipolar depression, p < .001 at T1, p < .0001 at T2), and BPRS activation factor score (mixed mania, p < .0001 at T1 and T2; bipolar depression, NS at T1, p < .01 at T2). Between-group comparisons revealed that patients with mixed mania showed significantly greater decrease in MADRS score (p < .001) and a greater proportion of responders (CGI-S) than patients with bipolar depression at endpoint (56% [N = 23] vs. 26% [N = 6], p = .02). Patients with mixed mania showed a greater reduction in suicidality, as measured by MADRS score, than patients with bipolar depression (p < .02). CONCLUSION: In our study, ECT was associated with a substantial reduction in symptomatology, in both patients with mixed mania and those with bipolar depression. However, the mixed mania group exhibited a more rapid and marked response as well as a greater reduction in suicidal ideation. Response to ECT was not influenced by the presence of delusions. PMID- 11488368 TI - Cognitive-behavioral management of patients with bipolar disorder who relapsed while on lithium prophylaxis. AB - BACKGROUND: The application of cognitive-behavioral treatment (CBT) to patients with bipolar disorder who had an affective episode while on lithium prophylaxis has received little research attention. The aim of this preliminary study was to test whether reduction of residual symptomatology by cognitive-behavioral methods could yield long-term beneficial effects in patients with bipolar disorder, as was found to be the case in recurrent unipolar depression. METHOD: Fifteen patients with RDC bipolar disorder, type I, who relapsed while on lithium prophylaxis despite initial response and adequate compliance were treated by cognitive-behavioral methods in an open trial. A 2- to 9-year follow-up was performed. RESULTS: Five of the 15 patients had a new affective episode during follow-up. CBT was associated with a significant reduction of residual symptomatology. CONCLUSION: These preliminary results suggest that a trial of CBT may enhance lithium prophylaxis and improve long-term outcome of bipolar disorder. PMID- 11488369 TI - A prospective study of the paradoxical relationship between impulsivity and lethality of suicide attempts. AB - BACKGROUND: Biological studies suggest that lower serotonergic activity is associated with both greater suicide lethality and impulsive personality traits. These results may lead to the conclusion that impulsivity in the attempt should be associated with greater lethality. However, Klerman's review of epidemiologic suicide studies suggests an inverse relationship between impulsivity and lethality. This seemingly paradoxical relationship between impulsivity and lethality has not been explored in large representative clinical samples of suicide attempts. METHOD: During 1996 to 1998, 478 individuals who attempted suicide were studied in a general hospital in Madrid, Spain. Impulsivity was measured as described in the literature by combining 2 items of Beck's Suicidal Intent Scale (active preparation for attempt and degree of premeditation). Lethality of the attempt was assigned 1 of 4 levels according to the need for medical and/or psychiatric treatment. RESULTS: More than half of the attempts were impulsive (55%; 95% confidence interval [CI], 51% to 59%), approximately one fourth of the attempts had an intermediate level of impulsivity (28%; 95% CI, 24% to 32%), and approximately one sixth of the attempts were not impulsive (17%, 95% CI, 13% to 21%). There was an inverse association between the impulsivity and lethality of the suicide attempt (chi2 = 62.639, df = 6, p < .0001). The most impulsive attempts tended to result in less morbidity, while the less impulsive attempts tended to be more lethal. CONCLUSION: If the inverse relationship between impulsivity and lethality is replicated in other large and representative samples, new studies will be needed to clarify the complex interactions between the clinical dimensions (lethality, impulsivity as a state, and impulsivity as a personality trait) and the biological correlates (particularly serotonergic function) of suicidal behavior. PMID- 11488370 TI - Effectiveness and safety of long-term antidepressant treatment in bipolar disorder. AB - OBJECTIVE: We sought to review research on use of antidepressants for long-term treatment of bipolar depression. METHOD: We conducted a computerized literature search of the MEDLINE, HealthStar, Current Contents, PsychInfo, and National Library of Medicine databases to identify studies involving antidepressant, anticonvulsant, or lithium use in bipolar disorder or manic-depressive illness published from 1966 through 2000. RESULTS: Only 7 blinded, controlled trials of long-term antidepressant treatment in bipolar disorders were found. The available information is not adequate to support the safety or effectiveness of long-term antidepressant treatment for bipolar depression, with or without mood-stabilizing cotherapy. CONCLUSION: Antidepressant treatment of bipolar depression is extraordinarily understudied. Controlled trials comparing specific antidepressants, particularly to compare mood-stabilizing agents given alone and combined with an antidepressant, are needed. PMID- 11488371 TI - Remission of SSRI-induced akathisia after switch to nefazodone. PMID- 11488372 TI - Chronic assaultive behavior improved with sleep apnea treatment. PMID- 11488373 TI - Negative symptoms of schizophrenia are improved by paroxetine added to neuroleptics: a pilot study. PMID- 11488374 TI - Pisa syndrome due to a cholinesterase inhibitor (donepezil): a case report. PMID- 11488375 TI - Bipolar affective disorder and idiopathic pulmonary fibrosis. PMID- 11488376 TI - Fears, phobias, and preparedness: toward an evolved module of fear and fear learning. AB - An evolved module for fear elicitation and fear learning with 4 characteristics is proposed. (a) The fear module is preferentially activated in aversive contexts by stimuli that are fear relevant in an evolutionary perspective. (b) Its activation to such stimuli is automatic. (c) It is relatively impenetrable to cognitive control. (d) It originates in a dedicated neural circuitry, centered on the amygdala. Evidence supporting these propositions is reviewed from conditioning studies, both in humans and in monkeys; illusory correlation studies; studies using unreportable stimuli; and studies from animal neuroscience. The fear module is assumed to mediate an emotional level of fear learning that is relatively independent and dissociable from cognitive learning of stimulus relationships. PMID- 11488377 TI - The biological basis of speech: what to infer from talking to the animals. AB - Speech perception and production are uniquely human adaptations. The mechanisms and laws that these adaptations implicate are tuned to linguistic rather than general auditory phenomena, leading to the view that speech is special (SiS). Despite the progress made by proponents of SiS, a small but growing "auditorist" program critical of SiS conscripts nonhuman animals such as quail and chinchilla and, using discrimination tasks on speech stimuli, incorrectly infers a common mechanism from similar cross-species performance. The author argues that the auditorist's refutation project must demonstrate not just cross-species isomorphisms of behavior but also either a common biological mechanism or common functional organization. The refutation project does neither. The author argues further that the auditorist's appeal to the principle of parsimony fails. Respect for the total available evidence undermines auditorism and bolsters SiS. PMID- 11488378 TI - The time course of perceptual choice: the leaky, competing accumulator model. AB - The time course of perceptual choice is discussed in a model of gradual, leaky, stochastic, and competitive information accumulation in nonlinear decision units. Special cases of the model match a classical diffusion process, but leakage and competition work together to address several challenges to existing diffusion, random walk, and accumulator models. The model accounts for data from choice tasks using both time-controlled (e.g., response signal) and standard reaction time paradigms and its adequacy compares favorably with other approaches. A new paradigm that controls the time of arrival of information supporting different choice alternatives provides further support. The model captures choice behavior regardless of the number of alternatives, accounting for the log-linear relation between reaction time and number of alternatives (Hick's law) and explains a complex pattern of visual and contextual priming in visual word identification. PMID- 11488379 TI - Contingencies of self-worth. AB - Research on self-esteem has focused almost exclusively on level of trait self esteem to the neglect of other potentially more important aspects such as the contingencies on which self-esteem is based. Over a century ago, W. James (1890) argued that self-esteem rises and falls around its typical level in response to successes and failures in domains on which one has staked self-worth. We present a model of global self-esteem that builds on James' insights and emphasizes contingencies of self-worth. This model can help to (a) point the way to understanding how self-esteem is implicated in affect, cognition, and self regulation of behavior; (b) suggest how and when self-esteem is implicated in social problems; (c) resolve debates about the nature and functioning of self esteem; (d) resolve paradoxes in related literatures, such as why people who are stigmatized do not necessarily have low self-esteem and why self-esteem does not decline with age; and (e) suggest how self-esteem is causally related to depression. In addition, this perspective raises questions about how contingencies of self-worth are acquired and how they change, whether they are primarily a resource or a vulnerability, and whether some people have noncontingent self-esteem. PMID- 11488380 TI - Conflict monitoring and cognitive control. AB - A neglected question regarding cognitive control is how control processes might detect situations calling for their involvement. The authors propose here that the demand for control may be evaluated in part by monitoring for conflicts in information processing. This hypothesis is supported by data concerning the anterior cingulate cortex, a brain area involved in cognitive control, which also appears to respond to the occurrence of conflict. The present article reports two computational modeling studies, serving to articulate the conflict monitoring hypothesis and examine its implications. The first study tests the sufficiency of the hypothesis to account for brain activation data, applying a measure of conflict to existing models of tasks shown to engage the anterior cingulate. The second study implements a feedback loop connecting conflict monitoring to cognitive control, using this to simulate a number of important behavioral phenomena. PMID- 11488381 TI - Simulating a lesion in a basis function model of spatial representations: comparison with hemineglect. AB - The basis function theory of spatial representations explains how neurons in the parietal cortex can perform nonlinear transformations from sensory to motor coordinates. The authors present computer simulations showing that unilateral parietal lesions leading to a neuronal gradient in basis function maps can account for the behavior of patients with hemineglect, including (a) neglect in line cancellation and line bisection experiments; (b) neglect in multiple frames of reference simultaneously; (c) relative neglect, a form of what is sometime called object-centered neglect; and (d) neglect without optic ataxia. Contralateral neglect arises in the model because the lesion produces an imbalance in the salience of stimuli that is modulated by the orientation of the body in space. These results strongly support the basis function theory for spatial representations in humans and provide a computational model of hemineglect at the single-cell level. PMID- 11488382 TI - Counter model for word identification: reply to Bowers (1999). AB - The counter model (R. Ratcliff & G. McKoon, 1997) was designed to explain the normal processes of word identification and how they are influenced by a prior encounter with a word. The model accounts for the findings of word identification experiments in which words are flashed briefly. A crucial finding is that prior encounters with words typically lead to biases such that a previously encountered word is more likely to be given as a response. However, for low-frequency words, a prior encounter can improve overall performance (J. S. Bowers, 1999; E. M. Wagenmakers, R. Zeelenberg, & J. G. W. Raaijmakers, 2000). The authors show how the model can explain this result. Also, J. S. Bowers (1999) has claimed that some earlier data concerning dissimilar alternatives in forced-choice experiments that support the counter model are spurious, but the authors show that his claims are incorrect. In sum, the authors argue for a theoretical approach that offers a detailed description of the cognitive processes of word identification and explains performance across tasks, measures, and independent variables. PMID- 11488383 TI - Contingency, causation, and adaptive inference. AB - In contingency judgment tasks involving 2 event types, individuals weight the a and b cells of a 2 x 2 contingency table more than the c and d cells. Some theorists have argued that they can provide normative justifications for this weighting and that the weighting reflects simple heuristics that are adaptive in the real world. The authors show that, to avoid error, individual judgments about real contingencies should be more subtle than these supposedly adaptive heuristics allow. PMID- 11488384 TI - Factors controlling metal distributions in the surface sediments of the Erdek Bay, Sea of Marmara, Turkey. AB - A geochemical study of the Erdek Bay sediments in the Sea of Marmara, Turkey, was carried out to investigate the heavy metal distributions and sources. The bay has an average depth of 34 m and a two-layer water column stratification, with a pycnocline at 20 m. The distribution of "total" metal (Fe, Mn, Pb, Cu, Ni, Cr, Zn and Hg) concentrations, together with sequential selective extraction analyses, shows that the relatively high metal concentrations are mainly due to land-based natural inputs from the erosion products of mineralized zones and mafic ultramafic rocks delivered to the bay by two rivers from the south. However, some Pb, Zn and Cu occur in the Fe-Mn-oxyhydroxide and organic fractions, suggesting, in part, some anthropogenic inputs in addition to the predominantly natural inputs. The vertical distribution of metal/A1 ratios in a gravity core in the bay shows no significant change with depth, except for an early diagenetic enrichment at 0.7-1.6 m below sea floor (mbsf) above a 4750-3500-year-old sapropelic layer. This distribution is a further indication of the insignificant antropogenic metal pollution in the bay. PMID- 11488385 TI - A comparative assessment of contaminants in fish from four resacas of the Texas, USA-Tamaulipas, Mexico border region. AB - A recent survey of contaminant information for the Lower Rio Grande Valley (LRGV), Texas, has shown that little is known about contaminants and their impacts on biota of resacas (oxbows) along the US-Mexico border. In 1996, fish were collected from four resacas in the Texas-Tamaulipas border region to assess contaminant loadings and their impacts on fish and birds. Tissue residue concentrations in fish were analyzed and also compared to two histopathological bioindicators of unhealthy environmental conditions. Of the organochlorine insecticides measured, DDE was the most common and was present at relatively high concentrations (10 microg/g w/w) at some sites. DDE concentrations were nearly 20 times greater in fish from resacas in Texas than from resacas in Tamaulipas, although the limited sample sizes obtained precluded statistical comparisons. DDE concentrations in fish from the two Texas resacas were also greater than those reported in fish from nearby areas during the 1980s and 1990s. Most trace element concentrations were similar among resacas from Texas and Tamaulipas. Arsenic, however, was two to six times greater in fish from a downtown resaca in Matamoros than in fish from other resacas in Tamaulipas and Texas. The bioindicators, pigment accumulation, and macrophage aggregates (MAs), in general, reflected the contamination indicated by the tissue residues for each site. Overall, it appears that some resacas of the US-Mexico border region are contaminant sinks and could pose potential health or reproductive problems for fish and wildlife, and humans that consume fish from those sites. PMID- 11488386 TI - Assessment of health risk levels associated with terrestrial gamma radiation dose rates in Nigeria. AB - A decree on radiation safety and protection have been signed by the Federal Government of Nigeria that established a regulatory body to control the importation, application, transportation and disposal of radioactive materials in the environment. The fundamental function of the body is embodied on radiation protection of the country's population and the environment. However, the protection and assessment of any radiation pollution in the environment to a large extent is based on the knowledge of the baseline of radiation dose rate levels due to natural radionuclides. This study assesses the risks associated with terrestrial gamma radiation dose rate levels in some cities across the major geological formations of the country. Results showed that in the Northern region the number of individuals at risk of incurring cancer ranged between 0.25 and 3.25 yr with an average value of 1.46+/-0.45, while in the southwestern region it ranged between 0.76 and 5.50 yr(-1) with a mean value of 1.66+/-0.63 and in the southeastern region it was between 0.17 and 0.89 yr(-1) with a mean value of 0.35+/-0.14. Assuming uniform exposure for the entire population of 98.5 million, it has been estimated that about 160 individuals annually are at risk of incurring cancer due to exposure to terrestrial gamma radiation. PMID- 11488387 TI - Monitoring of environmental heavy metals in fish from Nasser Lake. AB - Lake ecosystems are, in particular, vulnerable to heavy metal pollution. Tilapia nilotica is one of the aquatic organisms affected by heavy metals. Therefore, heavy metals Co, Cr, Cu, Fe, Mn, Ni, Sr and Zn were determined in different tissues of T. nilotica (ages 1, 1.5, 2, 2.5 and 3 years), which include the muscle, gill, stomach, intestine, liver, vertebral column and scales, to assess the fish pollution with heavy metals. In addition, the study was extended to determine these elements in the aquatic plant (Najas armeta), sediment and water of Khor E1-Ramel in Nasser Lake (Egypt). The study showed that of all the fish parts, fish liver accumulated the highest levels of Cu and Zn. Manganese presented in the intestine and stomach in the highest concentration. Scales exhibited the highest levels of Co, Cr, Ni and Sr, while the gill and vertebral column contains the lowest level of the studied elements. Heavy metals in different parts of T. nilotica differ with the fish growth and extraction rate of these elements from sediment, aquatic plant and lake water. Heavy metals under study in the edible parts of the investigated fish were in the safety permissible levels for human uses. PMID- 11488388 TI - Measurements and analysis of criteria pollutants in New Delhi, India. AB - Ambient concentrations of carbon monoxide (CO), nitrogen oxides (NOx), sulfur dioxide (SO2), and total suspended particulates (TSP) were measured from January 1997 to November 1998 in the center of downtown [the Income Tax Office (ITO) located on B.S.G. Marg] New Delhi, India. The data consist of 24-h averages of SO2, NOx, and TSP as well as 8 and 24-h averages of CO. The measurements were made in an effort to characterize air pollution in the urban environment of New Delhi and assist in the development of an air quality index. The yearly average CO, NOx, SO2, and TSP concentrations for 1997 and 1998 were found to be 4810+/ 2287 and 5772+/-2116 microg/m3, 83+/-35 and 64+/-22 microg/m3, 20+/-8 and 23+/-7 microg/m3, and 409+/-110 and 365+/-100 microg/m3, respectively. In general, the maximum CO, SO2, NOx, and TSP values occurred during the winter with minimum values occurring during the summer, which can be attributed to a combination of meteorological conditions and photochemical activity in the region. The ratio of CO/NOx (approximately 50) indicates that mobile sources are the predominant contributors for these two compounds in the urban air pollution problem in New Delhi. The ratio of SO2/NOx (approximately 0.6) indicates that point sources are contributing to SO2 pollution in the city. The averaged background CO concentrations in New Delhi were also calculated (approximately 1939 microg/m3) which exceed those for Eastern USA (approximately 500 microg/m3). Further, all measured concentrations exceeded the US National Ambient Air Quality Standards (NAAQS) except for SO2. TSP was identified as exceeding the standard on the most frequent basis. PMID- 11488389 TI - Trace metal concentrations in marine macroalgae from different biotopes in the Aegean Sea. AB - The commonest species of red, brown, and green macroalgae were sampled from a range of biotopes in the Aegean Sea and analysed for five different trace metals. Significant differences in metal concentrations were found among different seaweed species from the same biotope. The concentrations of metals in the various seaweed species may reflect their morphology, with those having a larger surface area having a greater internal content. Different species of seaweed have different affinities for different heavy metals. This may reflect competition between metals for binding or uptake sites in the seaweed. Comparing metal concentrations in algae among the studied sampling stations clearly indicates that the degree of accumulation depends not only on human activities but also on the geology of the specific area. While seaweed can be used successfully to assess the levels of heavy metals in the marine environment, not all elevated concentrations of heavy metals necessarily reflect increased levels of pollution. Indeed, the high concentrations of certain metals, e.g., Ni, found in our seaweed samples reflected the metaliferrous nature of the rock. It is therefore important to take account of a region's geology before attempting to interpret the data. PMID- 11488390 TI - Environmental challenges in computer manufacturing. AB - The purpose of this research is to examine the environmental policies and programs that have been instituted by four large computer manufacturers. These environmental concerns, while not highly publicized, present an extremely important challenge to computer manufacturers, as well as to our society in general. Though the environmental programs of these computer manufacturers have been in place for several years, the topic of environmental health and safety (EHS) in computer manufacturing is still a relatively new one. Four major computer manufacturers that have instituted environmental programs are selected in this research. Their environmental policies and programs are examined in detail and compared with each other to show the relative strength and weakness. The result of this research highlights that with the amazing growth of computer use for both business and in our daily life, it will only be a matter of time before the issue -- the environmental challenges in computer manufacturing -- gains prominence and exposure in our society on a large-scale basis. PMID- 11488391 TI - PAH emission from the incineration of three plastic wastes. AB - A batch-type, controlled-air incinerator was used for the treatment of polyvinyl chloride (PVC), high-density polyethylene (HDPE), and polypropylene (PP) plastic wastes. The concentration and composition of 21 individual polycyclic aromatic hydrocarbons (PAHs) in the raw wastes, flue gas (gas and particle phases), and ash were determined. Stack flue-gas samples were collected by a PAH stack sampling system. Twenty-one individual PAHs were analyzed primarily by a gas chromatograph/mass spectrometer (GC/MS). The CO concentration correlated well with the total PAH (R2 > .89), and thus can be used as a surrogate indicator for PAH emission. Excess amounts of air supply in the incineration of plastic wastes could decrease not only the concentration of the PAHs in the bottom ash but also the emission factor (EF) of the total PAH in the stack flue gas. Of the three plastic wastes, HDPE was found to have the highest mean EF of the total PAHs (462.3 mg/kg waste) from the stack flue gas. Incinerating PVC would result in a higher EF of PAHs (195.4 mg/kg waste) in the bottom ash. When PVC plastic wastes were incinerated, higher-ringed PAHs constituted a larger percentage in the bottom ash as compared to those from PP and HDPE plastics. By judging the output and input (O/I) ratio of the PAHs from the incineration trials of plastic wastes, the PAHs involved in incineration of three plastic wastes were almost entirely destroyed; and a low residual amount between 0.00018 and 0.00032 remained in the emission. PMID- 11488392 TI - Seasonal assessment of environmental tobacco smoke and respirable suspended particle exposures for nonsmokers in Bremen using personal monitoring. AB - The study was designed to determine seasonal differences in personal exposures to respirable suspended particles (RSP) and environmental tobacco smoke (ETS) for nonsmokers in Bremen, Germany. The subjects were office workers, either living and working in smoking locations or living and working in nonsmoking locations. One hundred and twenty four randomly selected nonsmoking subjects collected air samples close to their breathing zone by wearing personal monitors for 24 h or, in some cases, for 7-day periods during the winter of 1999. The investigation was repeated in the summer with 126 subjects, comprised of as many of the studied winter population (89 subjects) as possible. Saliva cotinine analyses were undertaken to verify the nonsmoking status of the subjects. Subjects wore one personal monitor while at work and one while away from the workplace on weekdays, and a third monitor at the weekend. Collected air samples were analysed for RSP, nicotine, 3-ethenylpyridine (3-EP) and ETS particles. The latter were estimated using ultraviolet absorbance (UVPM), fluorescence (FPM) and solanesol (SolPM) measurements. ETS exposure was consistently higher in the winter than in the summer, this pattern being particularly evident for subjects both living and working with smokers. The highest median 24-h time weighted average (TWA) concentrations of ETS particles (SolPM, 25 microg m(-3)) and nicotine (1.3 microg m(-3)) were recorded for subjects performing weekday monitoring during the winter. These were significantly higher than equivalent levels of ETS particles (SolPM, 2.4 microg m(-3)) and nicotine (0.26 microg m(-3)) determined during the summer. There were no appreciable differences between winter and summer percent workplace contributions to median TWA ETS particle and nicotine weekday concentrations, the workplace in Bremen, in general, contributing between 35% and 61% of reported median concentrations. Workers, on average, spent one-third of their time at work during a weekday, indicating that concentrations were either comparable or higher in the workplace than in the home and other locations outside the workplace. Median 24-h weekend ETS particle and nicotine concentrations for smoking locations were not significantly different from equivalent weekday levels during the winter, but were significantly lower during the summer. Based upon median 24-h TWA SolPM and nicotine concentrations for the winter, extrapolated to 1 year's ETS exposure, those subjects both living and working in smoking locations (the most highly exposed group) would potentially inhale 13 cigarette equivalents/year (CEs/y). However, based on a similar extrapolation of summer measurements, the same group of subjects would potentially inhale between 1.3 and 1.9 CEs/y. The most highly exposed subjects in this study, based upon 90th percentile concentrations for those both living and working in smoking locations during the winter, would potentially inhale up to 67 CEs/y in the winter and up to 22 CEs/y in the summer. This clearly demonstrates that seasonal effects should be taken into account in the design and interpretation of ETS exposure studies. Air sampling over a 7-day period was shown to be technically feasible, and subsequent RSP, ETS particle and nicotine levels determined by 7-day monitoring were not found to be significantly different from equivalent levels determined by 24-h monitoring. However, the longer sampling period resulted in the collection of an increased quantity of analytes, which improved the limits of quantitation (LOQ) and allowed a more accurate determination of low level ETS exposure. This was reflected by a reduced percentage of data falling below the LOQ for 7-day monitoring compared with 24-h monitoring. The use of a liquid chromatographic method with tandem mass spectrometric detection for saliva cotinine measurement afforded a greatly improved LOQ and greater accuracy at low concentrations compared with the radioimmunoassay (RIA) method used in previous studies by these authors. In this study, 17 subjects out of 180 tested (9.4%) were found to have saliva cotinine levels exceeding the selected threshold of 25 ng ml(-1) used to discriminate between smokers and nonsmokers. PMID- 11488393 TI - PTEN: a newly identified regulator of neuronal differentiation. AB - Even though phosphorylation of phosphatidylinositols by phosphoinositide 3-kinase has an important and pervasive role in the nervous system, little is known about the phosphatases that reverse this reaction. Recently, such a phosphatase, PTEN, was cloned as a tumor suppressor for gliomas. We now know that PTEN is a tumor suppressor for many tumor types and is a phosphatidylinositol phosphatase specific for the 3-position of the inositol ring. PTEN is expressed in most, if not all, neurons and is localized in the nucleus and cytoplasm. PTEN is not evident in neural processes or synapses. PTEN is induced during neuronal differentiation and is required for survival of differentiating neuronal cells. In summary, PTEN is a regulatory molecule with multiple functions at multiple subcellular sites. Further studies are required to determine which downstream pathways are regulated by PTEN, by which mechanisms PTEN activity is regulated, which stimuli regulate PTEN activity, and why a molecule that inhibits several survival pathways is induced during neurogenesis. PMID- 11488394 TI - Molecular basis for the perception of pain. AB - It is perhaps presumptuous to talk about the molecular basis of a subjective sensation such as pain, but defined conformational changes in membrane proteins, controlled by a family of extra- and intracellular messenger molecules, are known to underlie the activation of sensory nerve terminals and the process of synaptic neurotransmission, which are necessary for pain perception. Furthermore, a subset of neurotransmission processes has a permissive, and possibly exclusive, role in pain perception. Clearly, the experience of pain in the clinical sense with all its affective components of unpleasantness and suffering cannot yet be fully understood in molecular terms, but the process of nociception, whereby the signal generated as a result of tissue damaging or potentially damaging peripheral stimuli reaches and evokes neuronal activity in the central nervous system, is becoming better characterized. Recent advances in neurobiology have given us insights that are already helping improve understanding of the events that lead to a patient experiencing pain and, it is hoped, will also lead to more successful treatment strategies. PMID- 11488395 TI - Reward signaling by dopamine neurons. AB - Dopamine projections from the midbrain to the striatum and frontal cortex are involved in behavioral reactions controlled by rewards, as inferred from deficits in parkinsonism, schizophrenia, and drug addiction. Recent experiments have shown that dopamine neurons are not directly modulated in relation to movements. Rather, they appear to code the rewarding aspects of environmental stimuli. They show short, phasic increases of activity following primary food and liquid rewards ("unconditioned stimuli") and conditioned, reward-predicting stimuli of visual, auditory, and somatosensory modalities. They also display smaller activation-depression sequences after stimuli resembling rewards and after novel or particularly intense stimuli. Rewards are only reported as far as they occur differently than predicted. According to learning theories, a "prediction error" message may constitute a powerful teaching signal for behavior and learning. The phasic reward message is different from the more tonic enabling function of dopamine that is deficient in Parkinson's disease, indicating that dopamine neurons subserve different functions at different time scales. Neurons in other brain structures, such as the striatum, orbitofrontal cortex, and amygdala, code the quality, quantity, and preference of rewards. The dopamine reward prediction error signal may cooperate with these reward perception signals during the learning and performance of behavioral reactions to motivating environmental stimuli. PMID- 11488396 TI - Human neocortical development: the importance of embryonic and early fetal events. AB - The identification of numerous genes involved in the development of the cerebral cortex has led to an increased interest in the early stages of corticogenesis, when the first postmitotic neurons migrate into the cortical plate to form the foundation of the adult cortex. However, the cellular substrate of gene expression in early human cortical development is widely unknown. This article analyzes the complex sequence of events in the differentiation of the preplate, the predecessor of the neocortex, and discusses the possible origin and migratory routes of the neuronal populations involved in the transition from preplate to cortical plate. The neuronal classes present in embryonic and early fetal stages are redefined in terms of their relationship with the Reelin-Dab1 signaling pathway whose integrity is essential for successful migration into the cortex. A timetable of developmental steps is provided, and the peculiarities of the preplate derivatives in the human brain, marginal zone, and subplate are discussed. The results presented here may contribute to a deeper understanding of the pathogenesis of migration disorders. PMID- 11488397 TI - The place of the thalamus in frontal cortical-basal ganglia circuits. AB - The thalamus has long been thought to convey subcortical information to the cortex. Indeed, models of basal ganglia function attribute the primary role for the thalamus to a simple relay of information processed in the basal ganglia to the cortex. The thalamic nuclear groups that are associated primarily with this function are the ventral anterior and ventral lateral nuclei and the mediodorsal thalamic nucleus. However, recent studies have shown that the corticothalamic projection is important for the dynamics of the thalamocortical processing. Furthermore, the relay nuclei that carry basal ganglia output to the cortex have recently been shown to project back to the basal ganglia directly. These two recent developments indicate a more dynamic role for the thalamus in basal ganglia information processing than a passive relay. PMID- 11488398 TI - Bridging areas of injury in the spinal cord. AB - There is a devastating loss of function when substantial numbers of axons are interrupted by injury to the spinal cord. This loss may be eventually reversed by providing bridging prostheses that will enable axons to regrow across the injury site and enter the spinal cord beyond. This review addresses the bridging strategies that are being developed in a number of spinal cord lesion models: complete and partial transection and cavities arising from contusion. Bridges containing peripheral nerve, Schwann cells, olfactory ensheathing glia, fetal tissue, stem cells/neuronal precursor cells, and macrophages are being evaluated as is the administration of neurotrophic factors, administered by infusion or secreted by genetically engineered cells. Biomaterials may be an important factor in developing successful strategies. Due to the complexity of the sequelae following spinal cord injury, no one strategy will be effective. The compelling question today is: What combinations of the strategies discussed, or new ones, along with an initial neuroprotective treatment, will substantially improve outcome after spinal cord injury? PMID- 11488399 TI - Mesial temporal lobe epilepsy: what have we learned? AB - Mesial temporal lobe epilepsy is the most common form of human epilepsy, and its pathophysiological substrate is usually hippocampal sclerosis, the most common epileptogenic lesion encountered in patients with epilepsy. The disabling seizures associated with mesial temporal lobe epilepsy are typically resistant to antiepileptic drugs but can be abolished in most patients by surgical treatment. Anteromesial temporal resection, therefore, is the most common surgical procedure performed to treat epilepsy, and stereotactically implanted intracerebral electrodes are required in some patients to localize the epileptogenic region. This clinical setting provides a large number of patients for invasive in vivo research with microelectrode and microdialysis techniques and in vitro research following surgical resection on a single epileptic disorder. Consequently, much has now been learned about the fundamental neuronal mechanisms underlying the epileptogenic properties of the human hippocampus in mesial temporal lobe epilepsy. Parallel reiterative studies in patients and animal models of this disorder indicate that enhanced inhibition, in addition to enhanced excitation, underlies the appearance of hypersynchronous neuronal discharges responsible for generating spontaneous seizures. Recent studies have elucidated what may be unique electrophysiological markers of epileptogenicity, which could have valuable diagnostic utility. Although basic research on mesial temporal lobe epilepsy may ultimately suggest novel approaches to treatment and prevention, attention must also be given to maximizing the application of available effective treatments. In particular, the safety and efficacy of surgical therapy has greatly improved in recent years, yet this alternative treatment remains seriously underutilized worldwide. An appropriate increase in referral of patients with this surgically remediable syndrome to epilepsy centers will not only relieve a great many patients of their disabling seizures and reduce the burden of epilepsy but will also provide increased opportunities for invasive research that could ultimately result in even more effective therapies or cures. PMID- 11488400 TI - Effect of bilirubin on toxicity induced by trifluoperazine, dibucaine and praziquantel to erythrocytes. AB - Unconjugated bilirubin (UCB), like trifluoperazine (TFP), dibucaine (DBC) and praziquantel (PZQ), induces erythrocyte morphological changes, lysis and lipid exfoliation. In the present study we determined whether TFP, DBC and PZQ toxicity to erythrocytes was potentiated or reverted by UCB. Human erythrocytes were either treated or non-treated with 34.2 micromol/L UCB for 10 min prior to the incubation with toxic concentrations of TFP (0.12 mmol/L), DBC (1.5 mmol/L) or PZQ (3.0 mmol/L), for 1 h (37 degrees C). Studies of toxic effects included morphological analysis of erythrocytes, evaluation of hemoglobin release and loss of membrane lipids. Although UCB has an echinocytogenic effect, its co-incubation with TFP or PZQ did not alter the stomatocytogenic effect of the drug but enhanced DBC-induced stomatocytosis. Cell fusion was a common feature in experiments with DBC. Injurious effect of DBC to erythrocytes was potentiated by UCB as manifested by a marked increase in hemolysis (171%, p<0.05), and in elution of membrane cholesterol (73%, p<0.01) and phospholipids (123%, p<0.01). In opposite, toxic events produced by TFP and PZQ to erythrocytes were not aggravated by UCB. Interestingly, UCB prevented the loss of membrane cholesterol by PZQ (-36%, p<0.01), as well as that of phospholipids by TFP (-28%, p<0.05). These findings indicate that UCB potentiates DBC injury to erythrocytes, while protects membrane lipid elution by PZQ and TFP. Therefore, the relation of the benefits and risks of the administration of DBC to jaundiced patients should be carefully considered. PMID- 11488401 TI - Inhibition of brain monoamine oxidase activity by the generation of hydroxyl radicals: potential implications in relation to oxidative stress. AB - Monoamine oxidase (MAO) is an enzyme involved in brain catabolism of monoamine neurotransmitters whose oxidative deamination results in the production of hydrogen peroxide. It has been documented that hydrogen peroxide derived from MAO activity represents a special source of oxidative stress in the brain. In this study we investigated the potential effects of the production of hydroxyl radicals (*OH) on MAO-A and MAO-B activities using mitochondrial preparations obtained from rat brain. Ascorbic acid (100 microM) and Fe2+ (0.2, 0.4, 0.8, and 1.6 microM) were used to induce the production of *OH. Results showed that the generation of *OH significantly reduced both MAO-A (85-53%) and MAO-B (77-39%) activities, exhibiting a linear correlation between both MAO-A and MAO-B activities and the amount of *OH produced. The reported inhibition was found to be irreversible for both MAO-A and MAO-B. Assuming the proven contribution of MAO activity to brain oxidative stress, this inhibition appears to reduce this contribution when an overproduction of *OH occurs. PMID- 11488402 TI - A new glycogen synthase activity ratio in skeletal muscle: effects of exercise and insulin. AB - It was recently reported that MnSO4 stimulates glycogen synthase-dependent glucose transfer from UDPglucose into trichloroacetic acid precipitable endogenous glycoproteins (GSMn(T)) in human muscle extracts. To determine the physiologic significance of this reaction, we compared a new GS activity ratio, GSMn(T)/GSH(E) (where GSH(E) represents the usual glucose transfer to ethanol precipitable exogenous glycogen by GS at 7.2 mM glucose 6-phosphate), with the generally used GSL(E)/GSH(E) ratio (where GSL(E) represents glucose transfer at 0.17 mM glucose 6-P concentration). Biopsies were obtained from the quadriceps femoris muscle of healthy subjects at rest, after 40 min of bicycle exercise at approximately 65% of maximal oxygen uptake and after isometric contraction at 2/3 maximal force to fatigue (approximately 1 min). GSMn(T)/GSH(E) increased from 0.012+/-0.002 at rest to 0.054+/-0.008 (P<0.01) after 40 min of bicycle exercise and the increase in GSMn(T) activity was strongly related to the decrease in endogenous glycogen (i.e.. increase in short-chain endogenous glycoproteins) (r=0.90; P<0.05). On the other hand, GSL(E)/GSH(E) did not change significantly after bicycle exercise (rest = 0.49+/-0.04; exercise = 0.58+/-0.08, P>0.05). GSMn(T)/GSH(E) increased from 0.010+/-0.001 at rest to 0.016+/-0.002 (P<0.05) after isometric exercise, whereas GSL(E)/GSH(E) decreased from 0.27+/-0.04 to 0.20+/-0.02 (P<0.05) under corresponding conditions. Last, insulin, which stimulates glycogen synthesis, also increased GSMn(T)/GSH(E) (1.8-fold, P<0.05), as well as GSL(E)/GSH(E) (1.4-fold, P<0.05), in isolated rat soleus muscle. These data indicate that GSMn(T)/GSH(E) is influenced by endogenous substrate availability and covalent modification. Therefore, GSMn(T)/GSH(E) ratio may prove to be a useful alternative to other GS activity ratios that only reflect changes in the phosphorylation state of GS. PMID- 11488403 TI - Dietary administration of citrus nobiletin inhibits azoxymethane-induced colonic aberrant crypt foci in rats. AB - The modifying effects of dietary feeding of a polymethoxyflavonoid nobiletin isolated from Citrus unshiu on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were investigated in male F344 rats. We also assessed the effects of nobiletin on cell proliferation activity of ACF using a monoclonal antibody MIB-5. Rats were given subcutaneous injections of AOM (15 mg/kg body weight) once a week for 3 weeks to induce ACF. They also received the experimental diet containing 0.01% or 0.05% nobiletin for 5 weeks, starting one week before the first dosing of AOM. AOM exposure produced 139 +/- 35 ACF/rat at the end of the study (week 5). Dietary administration of nobiletin caused significant reduction in the frequency of ACF: 70 +/- 15 (50% reduction, p<0.001) at a dose of 0.01% and 63 +/- 10 (55% reduction, p<0.001) at a dose of 0.05%. Nobiletin feeding significantly lowered MIB-5-index in ACF. Also, dietary administration of nobiletin significantly reduced prostaglandin E2 content in the colonic mucosa. These findings might suggest possible chemopreventive ability of nobiletin, through suppression of cell proliferating activity of ACF, in the development of ACF. PMID- 11488404 TI - Carbetapentane attenuates kainate-induced seizures via sigma-1 receptor modulation. AB - We examined the effects of a non-opioid antitussive, carbetapentane (CB) on kainic acid (KA)-induced neurotoxicity in rats. KA administration (10 mg/kg, i.p.) produced robust behavioral convulsions lasting 4 to 5 h. CB (12.5 and 25 mg/kg. i.p.) pretreatment consistently and in a dose-dependent manner reduced the KA-induced seizures, mortality, and marked loss of cells in regions CA1 and CA3 of the hippocampus. Consistently, CB pretreatment also significantly attenuated the KA-induced increase in Fos-related antigen immunoreactivity in the hippocampus. In contrast, pretreatment with the sigma-1 receptor antagonist BD1047 (1 and 2 mg/kg, i.p.) blocked, in a dose-related manner, the neuroprotection afforded by CB. These results suggest that CB provides neuroprotection against KA insult via sigma-1 receptor modulation. PMID- 11488405 TI - Evaluation of hepatic cytochrome P4502E1 in the species-dependent bioactivation of 4-vinylcyclohexene. AB - 4-Vinyl-1-cyclohexene (VCH), is converted by multiple forms of cytochrome P450 (CYP) to two monoepoxides (4-vinyl-1-cyclohexene 1,2-epoxide [VCH-1,2-epoxide], 4 vinyl-1-cyclohexene 7,8-epoxide [VCH-7,8-epoxide]), and 4-vinyl-1-cyclohexene diepoxide (VCD). A greater degree of formation of these epoxides by female B6C3F1 mice as compared to Fischer 344 rats correlates with the ovarian toxicity observed only in the mice. Understanding which isoforms of CYP are involved in VCH bioactivation will better explain the species-dependent ovotoxicity of VCH. Present studies focus on the role of CYP2E1, as this isoform is responsible for the bioactivation of several structurally related small molecular weight compounds, including 1,3-butadiene. Hepatic microsomes prepared from either mice or rats pretreated with the CYP inducer acetone demonstrated 2-fold increases in the formation of VCH-1,2-epoxide. However, incubations with microsomes from cyp2e1-deficient mice compared to those from wild type mice revealed no differences in the rates of bioactivation of VCH to the monoepoxides. Since repeated exposure to VCH is required for VCH-induced ovotoxicity, rodents were dosed with VCH for 5 or 10 d to observe effects on the hepatic concentration of CYP2E1 and/or associated activities. VCH pretreatment failed to increase the concentration of CYP2E1 or CYP2E1 activity in either species, as measured by immunoblotting analysis and p-nitrophenol hydroxylation. Based on these data, it is concluded that CYP2E1 does not play a role in the species differences between mice and rats in the bioactivation of VCH following repeated exposure to VCH. Other isoforms, such as those in CYP2A and CYP2B subfamilies, are likely involved in VCH bioactivation. PMID- 11488406 TI - Lecithinized copper, zinc-superoxide dismutase ameliorates ischemia-induced myocardial damage. AB - We have reported that lecithin-conjugated recombinant human Cu, Zn-superoxide dismutase (lecithinized SOD) has greater pharmacological potency than unmodified SOD through an increase in cell membrane affinity and half-life in plasma. Recently, ischemia or hypoxia alone has been suggested to result in increased superoxide anions, which lead to apoptosis in cardiomyocytes. We tested the effect of lecithinized SOD in reducing the infarct size following prolonged myocardial ischemia without reperfusion. Rats were subjected to a 24-h left coronary occlusion. Lecithinized SOD, unmodified SOD, free lecithin derivative or PBS was administered intravenously 30 min before coronary occlusion. SOD concentration of the heart, measured by ELISA, was higher in the lecithinized SOD treated group than in the other groups 24 h after administration. The infarct area ratio of the heart, assessed by TTC staining, in the lecithinized SOD treated group was significantly smaller than those of the other groups. Both TUNEL-positive cardiomyocytes and DNA laddering were attenuated in the ischemic area of the heart treated with lecithinized SOD. Single bolus administration of lecithinized SOD had a cardioprotective effect against ischemia without reperfusion in the rat model of acute myocardial infarction, possibly due to its sustained high tissue concentration. PMID- 11488407 TI - Involvement of a calcium-independent pathway in plasmin-induced platelet shape change. AB - Plasmin-induced platelet activation is considered to be a cause of reocclusion after thrombolytic therapy with plasminogen activators. However, little is known regarding its mechanism and regulation, particularly with respect to the initial step shape change. We here demonstrate that a Ca2+-independent pathway is involved in plasmin-induced human platelet shape change, and that Rho-kinase plays an important role in this pathway. When the increase in cytosolic Ca2+ was prevented by an intracellular Ca2+ chelator, 5,5'-dimethyl-BAPTA, plasmin-induced platelet shape change was partially inhibited but still occurred. In the presence of 5,5'-dimethyl-BAPTA, a specific Rho-kinase inhibitor, Y-27632, completely inhibited the shape change. Phosphorylation of myosin light chain, a key regulator of platelet shape change, was completely inhibited by Y-27632 in 5,5' dimethyl-BAPTA-treated platelets. Although plasmin caused tyrosine phosphorylation of the 80 kDa protein during the shape change, it did not seem to have a critical role. cAMP-elevating agents inhibited plasmin-induced shape change in 5,5'-dimethyl-BAPTA- or Y-27632-treated platelets with similar efficiency. These results indicated that plasmin causes platelet shape change by activating Ca2+-dependent and Ca2+-independent-Rho-kinase-dependent pathways, both of which are sensitive to cAMP. PMID- 11488408 TI - Glucocorticoid receptors in human peripheral blood mononuclear cells in relation to explosive performance in elite handball players. AB - Ten handball players, members of the Italian National Team (aged 20-25 years), were studied in two sessions corresponding to different performance levels. The first session occurred one week after the end of the regular season of the Italian Handball Federation: it corresponded to the beginning of the training cycle for the European Handball Championship. The second session occurred ten weeks after the first session. During this period, training consisted of 3 weeks of active recovery and 7 weeks of increasing workload. For each session, jumping performances (maximal height in a single jump, average mechanical power for a 15 s set of consecutive jumps) were evaluated. Venous blood samples were collected in resting conditions immediately before jumping performances to assess cortisol and testosterone plasma concentrations and glucocorticoid receptors (GcR) binding capacity and affinity in peripheral blood mononuclear cells (PBMCs). All the parameters, except GcR binding affinity, increased in the second session. The trends of variation in jumping performances, steroid hormone levels and GcR binding capacity were similar. For testosterone, this agrees with the hypothesis that an adequate level of this hormone is a prerequisite for improvement in explosive performances. For cortisol, higher GcR binding capacity after 10 weeks of training (with respect to initial values) indicated an up-regulation of GcR concomitant with the increase in hormone levels and performances. These findings suggest that the adaptation to training, confirmed by the improvement in performance, is characterized by a high value of GcR binding capacity and that it is mediated, among other factors, by the hormone levels and up-regulation of the receptors. PMID- 11488409 TI - Opiate-like substances mediate norepinephrine-induced but not serotonin-induced antinociception at spinal level: reevaluation by an electrophysiological model of formalin test in rats. AB - After subcutaneous injection of formalin (5%, 50 microl) into a hindpaw of rats, biphasic excitatory nociceptive discharges were recorded extracellularly in thalamic parafascicular neurons. Intrathecal (i.t.) administration of either norepinephrine (NE. 6 nmol, 10 microl) or serotonin (5-HT, 120 nmol, 10 microl) prior to the second phase significantly inhibited the second phase of the formalin-induced parafascicular nociceptive discharges. Intrathecal naloxone (Nal, 50 nmol, 10 microl) did not show any effect on the parafascicular nociceptive discharges. However, when i.t. Nal was given 5 min before NE, Nal prevented the NE antinociceptive effect. Pre-administration of Nal before 5-HT did not affect the antinociceptive effects of 5-HT on the second phase of nociceptive discharges. These results indicate that opiate-like substances are involved in the mediation of NE-induced antinociception. It is suggested that endogenous NE and 5-HT released from brainstem descending terminals at the spinal level carry out their antinociceptive actions differently. PMID- 11488411 TI - SWISS-PROT: connecting biomolecular knowledge via a protein database. AB - With the explosive growth of biological data, the development of new means of data storage was needed. More and more often biological information is no longer published in the conventional way via a publication in a scientific journal, but only deposited into a database. In the last two decades these databases have become essential tools for researchers in biological sciences. Biological databases can be classified according to the type of information they contain. There are basically three types of sequence-related databases (nucleic acid sequences, protein sequences and protein tertiary structures) as well as various specialized data collections. It is important to provide the users of biomolecular databases with a degree of integration between these databases as by nature all of these databases are connected in a scientific sense and each one of them is an important piece to biological complexity. In this review we will highlight our effort in connecting biological information as demonstrated in the SWISS-PROT protein database. PMID- 11488410 TI - Effect of 3,5,3'-triiodo-L-thyronine on amino acid accumulation and membrane potential in Sertoli cells of the rat testis. AB - The purpose of this study was to investigate the effect of T3 on amino acid accumulation and on the membrane potential of Sertoli cells of immature rat testes. Testes of pre-pubertal and pubertal rats were pre-incubated (30 min) in Krebs-Ringer bicarbonate buffer and incubated in the presence of [14C]methylaminoisobutyric acid with and without T3 for 15, 45 and 60 min. The hormone (10(-6) M and 10(-7) M) significantly stimulated amino acid accumulation in 6 and 13-day old rat testes but did not have any effect in neonatal and pubertal animals. T3 produced a dose-dependent hyperpolarizing effect at concentrations of 10(-6) M, 10(-5) M, 2 x 10(-5) M and 10(-4) M. We conclude that T3 induces a membrane hyperpolarization in Sertoli cells and stimulates amino acid accumulation in immature rat testes, demonstrating that the hormone has a rapid plasma membrane action. PMID- 11488412 TI - Degradation of mutant proteins, underlying "loss of function" phenotypes, plays a major role in genetic disease. AB - Many Mendelian monogenic disorders are caused by loss of the function of a single protein. This can result from rapid degradation of the mutant protein by cellular proteases, which reduces the steady-state concentration of the protein within the cell. The susceptibility of a protein to such proteolytic breakdown depends upon its kinetics of monomer folding and oligomer assembly and upon the intrinsic (thermodynamic) stability of its functional native-state conformation. Other cellular proteins, notably molecular chaperones, promote correct protein folding and assembly and thus provide some protection against degradation. An accumulation of recent evidence indicates that premature or accelerated degradation of mutant proteins, provoked by aberrations in their conformation, occurs in various subcellular compartments and represents a significant and prevalent pathogenic mechanism underlying genetic diseases. Inter-individual variability in proteolytic and folding systems can in part explain why "simple monogenic diseases" often display inconsistent genotype-phenotype correlations which show these disorders to be in reality quite complex. Protein folding and degradation may also be modulated artificially using exogenous small molecules. The identification or design of compounds which can interact specifically with particular target proteins, and which in so doing can exert beneficial effects on protein folding, assembly and/or stability, is beginning to open up a new and remarkably promising avenue for the treatment of diverse genetic disorders. PMID- 11488413 TI - DNA measurement and cell cycle analysis by flow cytometry. AB - Measurement of cellular DNA content and the analysis of the cell cycle can be performed by flow cytometry. Protocols for DNA measurement have been developed including Bivariate cytokeratin/DNA analysis, Bivariate BrdU/DNA analysis, and multiparameter flow cytometry measurement of cellular DNA content. This review summarises the methods for measurement of cellular DNA and analysis of the cell cycle and discusses the commercial software available for these purposes. PMID- 11488414 TI - Detection of human papillomavirus DNA in benign oral squamous epithelial lesions in Venezuela. AB - The polymerase chain reaction (PCR) was applied to the detection of human papillomavirus (HPV) infection in biopsies taken from clinically normal oral mucosa of 20 subjects and clinical lesions of 40 patients. PCR for HPV-DNA amplification was performed using consensus primers MYO9/MYO11 and subsequent typing for HPV of high and low oncogenic risk HPV types were identified by restriction enzyme analysis (restriction fragment length polymorphism, RFLP). The HPV viral genome was present in 55% (22/40) of the oral benign lesions (OBL) and in 10% (2/20) of the control samples. In the PCR+ OBL, we observed 90.9% of low oncogenic risk types (HPV-6 -13 and -32) and 9.1% of the samples had a mixed infection with low and high oncogenic types (HPV-6 and -16). In the control samples, we observed one patient with HPV-6 and another with HPV-6 and -16 in the same sample. All of the eight focal epithelial hyperplasia cases were positive for low risk HPV types (88% HPV-13 and 12.5% HPV-32). In conclusion, this study demonstrates a high incidence of HPV in oral benign lesions from Venezuelan patients. PMID- 11488415 TI - Epithelial-mesenchymal interactions in experimental oral mucosal carcinogenesis. AB - In an effort to come to a better understanding of human oral mucosal carcinogenesis, an animal model was used in which the carcinogen 4-nitroquinoline 1-oxide was applied to rat palatal mucosa for varying periods of time. Histological and histometric analyses showed that there were quantifiable differences in the palatal epithelium to which carcinogen had been applied in comparison with control tissue. Tissue recombination experiments, using various combinations of the palatal mucosa and analysed after recovery from transplantation to hypothymic BALB/c mice, showed that control epithelium recombined with connective tissue from carcinogen-treated mucosa was altered, indicating that the underlying connective tissue modified histomorphological aspects of the epithelium in the later stages of carcinogenesis. PMID- 11488416 TI - Absence of leukocyte microchimerism in oral lichen planus (OLP): an in situ hybridisation study. AB - Oral lichen planus (OLP) is a relatively common chronic inflammatory disease. The majority of patients are between 30 and 50 years of age with a higher incidence in females. The aetiology is unknown and various hypotheses on the pathogenic mechanisms, including autoimmunity, have been proposed over the years. In the present study, we investigated whether leukocyte microchimerism, a biological situation implicated in the aetiology of some autoimmune diseases, might play a role in the pathogenesis of OLP. We used in situ hybridisation to identify Y chromosome DNA in a series of formalin-fixed paraffin-embedded oral mucosa biopsies of women with established clinical and histological disease who had given birth to a male child. The positive control, two mucosal specimens from a man with OLP, showed over 90% of keratinocytes and cells within the inflammatory infiltrate, a positive nuclear signal. The negative control, biopsies from three women having carried only female foetuses and one nulliparous woman, all with OLP, did not show any nuclear signal. In the fifteen selected cases of OLP biopsies from women who had only male offspring, nucleated cells containing the Y chromosome were not detected within the chronic inflammatory infiltrate. These results suggest that unlike some other immunologically mediated diseases, leukocyte microchimerism does not seems to be involved in the pathogenesis of OLP. PMID- 11488417 TI - Some specific human leukocyte antigen (HLA)-DR/DQ haplotypes are more important than individual HLA-DR and -DQ phenotypes for the development of mucocutaneous type of Behcet's disease and for disease shift from recurrent aphthous stomatitis to mucocutaneous type of Behcet's disease. AB - The phenotype and haplotype frequencies of human leukocyte antigens (HLA)-DR and DQ in 32 Chinese patients with the mucocutaneous (MC) type of Behcet's disease (BD) were calculated and compared with those in 310 healthy control Chinese and with those in 80 Chinese patients with recurrent aphthous stomatitis (RAS). We found that the phenotype frequency of HLA-DRw8 [corrected P (Pc)<0.005] and the haplotype frequencies of HLA-DRw8/DQw1 (Pc<0.005), -DRw8/DQw5(w1) (Pc<0.0005), DRw12(5)/DQw1 (Pc<0.005), -DRw12(5)/DQw6(w1) (Pc< 0.0005), and -DRw52/DQw1 (Pc<0.005) in patients with the MC type of BD were significantly greater than those in healthy control subjects. This finding suggests that individual Chinese with HLA-DRw8 antigen and HLA-DRw8/DQw1, -DRw8/DQw5(w1), -DRw12(5)/DQw1, DRw12(5)/DQw6(w1) and -DRw52/DQw1 haplotypes are more likely to have the MC type of BD. Furthermore, the relative risks (RRs) of HLA-DRw8/DQw1 (5.6), -DRw8/ DQw5 (w1) (10.0), and -DRw12(5)/DQw6(w1) (14.4) haplotypes in patients with the MC type of BD were equal to or higher than the RR of HLA-DRw8 phenotype (5.6), suggesting that some of the HLA-DR/DQ haplotypes may play more important roles than the individual HLA-DR and -DQ phenotypes for the development of the MC type of BD. The phenotype frequencies of HLA-DR5 (Pc<0.01), -DRw8 (Pc<0.005) and -DQw1 (Pc<0.05) as well as the haplotype frequencies of HLA-DR5/DQw1 (P<0.005) and DRw8/DQw1 (Pc<0.00005) in patients with the MC type of BD were significantly higher than those in patients with RAS. Moreover, the RRs of HLA-DR5/DQw1 (29.1) and -DRw8/DQw1 (47.4) haplotypes were greater than the RRs of HLA-DR5 (10.4), DRw8 (23.4) and -DQw1 (4.0) antigens. These results suggest that some specific HLA-DR/DQ haplotypes may be more important than the individual HLA-DR and -DQ phenotypes in the disease shift from RAS to the MC type of BD. PMID- 11488418 TI - Reproducibility of biopsy grade in Sjogren's syndrome. AB - The purpose of this study was to examine the reproducibility of biopsy grades at various tissue depths in Sjogren's syndrome. The biopsy grades of 38 minor salivary gland biopsies were examined at 6 microm, 50 microm, 100 microm, 150 microm, 200 microm, and 250 microm tissue depths. Tissue sections were stained with routine hematoxylin and eosin, graded I-IV, and compared with the initial "baseline" biopsy grade. The majority of the biopsies showed a wide range of grade variability at all depths. No tissue depth was consistently reproducible for any grade (P> or =0.41, 0.64, 0.91, and 0.20, respectively). The difference between baseline grades and grades of deeper sections was sufficient to impact the diagnosis of Sjogren's syndrome in approximately 60% of the biopsies (P<0.001). The overall result of this study suggests that examination of multiple sections of minor salivary gland biopsies is advisable to improve the reliability of the grade when evaluating Sjogren's syndrome. PMID- 11488419 TI - Immunohistochemical localization of members of the transforming growth factor (TGF)-beta superfamily in normal human salivary glands and pleomorphic adenomas. AB - Although pleomorphic adenoma is the most common type of salivary gland epithelial tumor, it frequently contains "mesenchymal"-like components, including myxoid or chondroid tissues. We reported previously that chondroid tissue formation in pleomorphic adenoma was associated with overexpression of bone morphogenetic proteins (BMPs) by neoplastic myoepithelial cells. BMPs belong to the transforming growth factor (TGF)-beta superfamily, so we hypothesized that pleomorphic adenoma may express TGF-betas and that these molecules may regulate mesenchymal-like tissue formation. To evaluate this hypothesis, we immunohistochemically examined TGF-beta1, -beta2 and -beta3 expression and localization in normal salivary glands and in 43 cases of pleomorphic adenomas. There was no evidence of TGF-beta1 expression in normal salivary glands or pleomorphic adenomas. Signals for TGF-beta2 in the normal salivary glands were observed in the intercalated ducts, while in pleomorphic adenomas they were observed in the inner ductal cells of the tubulo-glandular structures. Signals for TGF-beta3 in the normal salivary glands were observed in mucous cells, whereas in pleomorphic adenomas they were observed in the solid nests of neoplastic myoepithelial cells, in the portion showing squamous metaplasia, and in the inner ductal cells of tubulo-glandular structures. TGF-betas induce ectopic cartilage formation in vivo, but chondroid tissues in pleomorphic adenomas showed only weak TGF-beta3 expression. TGF-beta may be related to differentiation of the inner ductal cells and the neoplastic myoepithelial cells. In conclusion, pleomorphic adenomas expressed TGF-beta2 and -beta3, which may be associated with differentiation of the inner ductal cells and neoplastic myoepithelial cells. PMID- 11488420 TI - Discrimination of ameloblastomas from odontogenic keratocysts by cytokine levels and gelatinase species of the intracystic fluids. AB - In the present study, we measured the levels of the cytokines and gelatinase species in the fluids of ameloblastomas and odontogenic keratocysts, and showed that ameloblastomas can be distinguished from odontogenic keratocysts by the use of these biochemical data. We found that interleukin (IL)-1alpha and IL-1beta levels in the intracystic fluids of ameloblastomas were significantly lower than those in the fluids of odontogenic keratocysts, while IL-6 levels in the fluids of ameloblastomas were significantly higher than those in the fluids of odontogenic keratocysts. On the other hand, no significant differences in tumor necrosis factor (TNF)-alpha levels of the fluids were detected between ameloblastomas and odontogenic keratocysts. An immunohistochemical study revealed that the staining intensity of IL-1alpha, IL-1beta and TNF-alpha in the tumor cells of ameloblastomas was significantly weaker than that in the epithelial cells of odontogenic keratocysts, while the staining intensity of IL-6 in the tumor cells was significantly stronger than that in the epithelial cells of odontogenic keratocysts. Gelatin zymography of the fluids showed that only a small amount of pro-MMP-9 was detected in ameloblastomas, while both pro-MMP-9 and the active form of MMP-9 were detected in 8 of 10 cases of odontogenic keratocysts. Thus, ameloblastomas can be distinguished from odontogenic keratocysts by measuring IL-1alpha and IL-6 levels, and gelatinase species in the fluids. PMID- 11488421 TI - Immunohistochemical localization of fibroblast growth factors FGF-1 and FGF-2, and receptors FGFR2 and FGFR3 in the epithelium of human odontogenic cysts and tumors. AB - Acidic (FGF-1) and basic (FGF-2) fibroblast growth factors are members of a family of growth factors that function in growth, differentiation and regeneration of a variety of tissues. Their presence in human odontogenic cysts and tumors has not been previously investigated. This study was designed to detect immunohistochemically the presence of these factors and two fibroblast growth factor receptors (FGFR2 and FGFR3) in a cross section of odontogenic cysts and tumors, to determine if they may be involved in the differentiation of odontogenic epithelium or, more specifically, in the development of particular cysts or tumors. Archival formalin-fixed paraffin-embedded tissues were used. With some exceptions, FGF-2 and the receptor FGFR2, were found in the cytoplasm and occasionally in the nuclei of cells of odontogenic epithelium, while FGF-1 and the receptor FGFR3, were absent or only focally or weakly detected, using standard immunohistochemical techniques. The data are similar to those published for normal murine odontogenesis, suggesting that these factors are associated with odontogenic differentiation rather than pathogenesis. The presence of significant nuclear staining in odontogenic epithelium associated with embryonic mesenchyme in ameloblastic fibromas and ameloblastic fibro-odontomas suggests that FGF-2 may be involved in directing nuclear activity at the histodifferentiation stage of odontogenesis. PMID- 11488422 TI - Immunohistochemical analysis of cell-cycle- and apoptosis-related factors in lining epithelium of odontogenic keratocysts. AB - We examined the immunohistochemical expressions of cell-cycle- and apoptosis related factors to investigate the possible role of these factors in odontogenic keratocyst (OKC). Expression of cyclin D1 and p16 protein was detected in the basal and parabasal cells in lining epithelium of OKCs and was found more frequently in basal cell nevus syndrome (BCNS)-associated OKCs than in primary or recurrent OKCs. Positivity for p21 protein was detected in basal to superficial cells, whereas that for p27 protein was detected in parabasal to superficial cells in lining epithelium of OKCs. DNA topoisomerase IIalpha reacted with nuclei in basal and parabasal cells of the lining epithelium of OKCs, and positive cells were observed in BCNS-associated OKCs significantly more frequently than in primary or recurrent OKCs. Expression of Fas in suprabasal to superficial cells and expression of Fas-L in basal and parabasal cells were detected in lining epithelium of OKCs. Immunoreactivity for caspase-3 was detected in basal to suprabasal or superficial cells in lining epithelium of OKCs. Single stranded (ss)DNA-positive nuclei were detected in superficial cells in lining epithelium of OKCs. Fas was more broadly distributed in BCNS-associated OKCs than in primary OKCs, and ssDNA-positive cells were observed in BCNS-associated OKCs significantly more frequently than in primary or recurrent OKCs. These results suggest that BCNS-associated OKCs might be a distinguishable entity from solitary OKCs. PMID- 11488423 TI - A case of submandibular malignant rhabdoid tumor transformed from papillary thyroid carcinoma. AB - Malignant rhabdoid tumor (MRT) in the neck region is very rare. We report a case of MRT in a 60-year-old woman who had a history of papillary carcinoma of the thyroid gland 7 years previously. One year before admission, in 1995, thyroid carcinoma recurred, and the tumor contained a small undifferentiated region with rhabdoid features. The tumor in 1996 consisted of round to oval rhabdoid cells with abundant cytoplasm, and the growth pattern was diffuse and infiltrative, with no papillary structures. We therefore concluded that the lesion was MRT, transformed from papillary thyroid carcinoma. PMID- 11488424 TI - First Asian ISSAM Meeting on the Aging Male, Kuala Lumpur, Malaysia, 1-3 March 2001--an overview. PMID- 11488426 TI - Drugs for the treatment of allergic diseases. AB - Many kinds of drugs are used for the treatment of allergic diseases. Glucocorticoids are the most efficacious drugs and widely used for the treatment of allergic diseases. Recently, effectiveness of inhaled glucocorticoids for the treatment of bronchial asthma has been established. Beclomethasone dipropionate and fluticasone propionate, which are degraded easily after absorption, are applied by inhalation. Histamine is one of the most important mediators in allergic reactions and antihistamines have widely been applied for the treatment of allergic skin diseases. In Japan, over 20 antiallergic drugs, such as mediator release inhibitors, mediator antagonists and mediator synthesis inhibitors, have been developed. Recently developed compounds such as pranlukast and suplatast are very effective. To relieve the asthmatic attack, bronchodilators such as beta2 adrenoceptor agonists, theophylline and anti-cholinergic drugs are used. Clinical application of tacrolimus ointment has just started for the treatment of atopic dermatitis. Recently the number of allergic patients has increased. The onset and development of allergic diseases are considered to be dependent on both the genetic factors and the environmental factors. For the successful treatment of patients with allergic diseases, it is also important to consider the control of environmental factors. PMID- 11488425 TI - Adenosine, oxidative stress and cytoprotection. AB - Adenosine, a metabolite of ATP, serves a number of important physiological roles in the body. These actions contribute to sedation, bradycardia, vasorelaxation, inhibition of lipolysis and regulation of the immune system and are mediated, in part, through activation of three distinct adenosine receptor (AR) subtypes. To date, four receptor types have been cloned: A1, A2A, A2B and A3. It is becoming increasing clear that adenosine contributes significantly to cytoprotection, a function mediated principally by the A1AR and A3AR. In this review, we survey the literature on the role of adenosine and the mechanisms underlying cytoprotection and ischemic preconditioning, a process characterized by cytoprotection derived from repeated brief ischemic challenges. An important recent observation is that the expression of several AR subtypes could be regulated by oxidative stress to provide a greater cytoprotective role. Thus, like other proteins known to be regulated during ischemia, the A1AR and A3AR can be considered as being inducible receptors. PMID- 11488427 TI - Effects of YM905, a novel muscarinic M3-receptor antagonist, on experimental models of bowel dysfunction in vivo. AB - We investigated the effects of YM905 [(+)-(1S,3'R)-quinuclidin-3'-yl 1-phenyl 1,2,3,4-tetrahydroisoquinoline-2-carboxylate monosuccinate], a new orally active muscarinic M3-receptor antagonist, on bowel dysfunction in vivo using experimental models that reproduce the symptoms present in irritable bowel syndrome (IBS). YM905 potently inhibited restraint stress-induced fecal pellet output in fed rats (ED50: 4.0 mg/kg) and diarrhea in fasted rats (ED50: 1.7 mg/kg), with similar potencies to the inhibition of bethanechol-, neostigmine- and nicotine-induced fecal pellet output in rats (ED50: 3.3, 7.9 and 4.5 mg/kg, respectively). YM905 also inhibited 5-hydroxytryptamine (5-HT)-, prostaglandin E2 and castor oil-induced secretory diarrhea in mice (ED50: 5.5, 14 and 6.3 mg/kg, respectively), but showed no significant effect on cholera toxin-induced intestinal secretion in mice. In addition, YM905 (3, 10 mg/kg) reversed morphine decreased postprandial defecation in ferrets, a model of spastic constipation, whereas remosetron, a 5-HT3-receptor antagonist, was not effective. The mode of YM905 action was similar to that of darifenacin, a selective M3-receptor antagonist, with equivalent potencies. By contrast, propantheline, an antimuscarinic drug that has been used for IBS, was much less potent. These results show that YM905 ameliorates a wide spectrum of bowel dysfunctions through the blockade of M3 receptors, suggesting its therapeutic potential for treating IBS. PMID- 11488428 TI - Effects of fluvastatin and its major metabolites on low-density lipoprotein oxidation and cholesterol esterification in macrophages. AB - We investigated effects of fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, and its major metabolites, M2 and M4, on CuSO4 induced low-density lipoprotein (LDL) oxidation and cholesteryl ester accumulation in mouse peritoneal macrophages. All the test compounds inhibited LDL oxidation, and M2 had the most potent effect comparable to vitamin E. When LDL was previously incubated with the test compounds in the presence of CuSO4, the pre-treatment resulted in a marked reduction of facilitated cholesteryl ester accumulation in macrophages. Supplementation of mevalonate did not overcome the inhibitory effects of fluvastatin and its metabolites on both LDL oxidation and facilitated cholesterol esterification. Pravastatin, another HMG-CoA reductase inhibitor, did not show any inhibitory effect. Consequently, these effects of fluvastatin and its metabolites are considered to be derived from their own unique chemical structures. Moreover, fluvastatin and M2 directly inhibited cholesterol esterification induced by oxidized LDL in macrophages, but pravastatin was also found to have a weak effect. As their inhibitory effects were overcome by addition of mevalonate, the direct inhibitory effect on cholesterol esterification would be a common property of HMG-CoA reductase inhibitors. The inhibitory effects of fluvastatin and its metabolites on both LDL oxidation and cholesterol esterification in macrophages may contribute to the antiatherogenic action in vivo. PMID- 11488429 TI - Effects of L-arginine on penicillin-induced epileptiform activity in rats. AB - It has been suggested that nitric oxide (NO) is involved in the pathophysiology of epilepsy. Data are, however controversial because it is not clear whether NO has pro- or anticonvulsant effects. The aim of this study was to investigate the effects of NO on penicillin G-induced epileptiform activity. The left cerebral cortex was exposed by craniotomy in urethane-anesthetized Wistar rats. The epileptic activity was produced by intraperitoneal injection of penicillin G (3 million U/kg, i.p.). The ECoG (electrocorticogram) activity was displayed on a four-channel recorder. At 39.7 +/- 5.4 min after penicillin administration, large amplitude sharp waves appeared in the ECoG. Mean spike frequency and mean spike amplitude were calculated as 29.5 +/- 3.2/min and 865 +/- 91 microV, respectively, at the 55th min. 7-Nitroindazole (60 mg/kg, i.p.) injection 30 min before penicillin G administration significantly reduced the latency of epileptiform activity. Intracerebroventricular administration of L-arginine (300 microg/2 microl, i.c.v.) and sodium nitroprusside (100 microg/2 microl, i.c.v.) suppressed epileptiform activity. Saline (2 microl) and D-arginine (300 microg/2 microl, i.c.v.) administration into the cerebral ventricle were completely ineffective on epileptiform activity (P<0.01). These findings suggest that NO may be an endogenous antiepileptic substance. PMID- 11488430 TI - Mechanism of 7,12-dimethylbenz[a]anthracene-induced immunotoxicity: role of metabolic activation at the target organ. AB - The polycyclic aromatic hydrocarbon, 7,12-dimethylbenz[a]anthracene (DMBA), is an immunosuppressor as well as a potent organ-specific carcinogen. To understand the organ-specific mechanism of DMBA-induced lymphoid toxicity, aryl hydrocarbon nonresponsive mice and microsomal epoxide hydrolase (mEH)-null mice were analyzed. DMBA caused a dose-dependent decrease in spleen weights, but not the thymus weights in aryl hydrocarbon-nonresponsive mice. On the other hand, both spleen and thymus weights were decreased to less than a half in wild-type mice exposed to 30 mg/kg of DMBA. In contrast, no decrease was detected in spleen weights of mEH-null mice exposed to up to 100 mg/kg of DMBA, while thymus weights were markedly lower. Responses to the B-cell mitogen lipopolysaccharide and to T cell mitogen phytohemagglutinin were nearly completely abolished in splenocytes isolated from wild-type mice treated with 100 mg/kg of DMBA. These responses were decreased, but maintained in splenocytes isolated from mEH-null mice treated with DMBA. Two DMBA metabolites dependent on mEH including DMBA-3,4-diol were detected in an HPLC chromatogram of spleen microsomes isolated from wild-type mice, but not those from mEH-null mice. These results suggest the involvement of mEH in splenic activation of DMBA for immunotoxicity and the difference for the DMBA induced lymphoid toxicity between spleen and thymus. PMID- 11488431 TI - Contribution of chloride channel activation to the elevated muscular tone of the pulmonary artery in monocrotaline-induced pulmonary hypertensive rats. AB - In monocrotaline-treated rat pulmonary artery from which endothelium was removed, greater spontaneous muscular tone was observed under resting conditions than in vehicle-treated artery. The aim of the present study was to show the possible contribution of Cl- channels in the mechanism of the elevated tone. Verapamil almost completely inhibited the elevated spontaneous muscular tone by decreasing [Ca2+]i. The elevated muscular tone was also inhibited by 4,4'-diisothiocyanato stilbene-2,2'-disulfonic acid (DIDS), a Cl- channel inhibitor. After the inhibition of muscular tone by DIDS, verapamil did not induce further relaxation. Quantitative RT-PCR analysis indicated that the mRNA levels of ClC3 and Ca2+ activated Cl- channels did not change in the pulmonary hypertensive pulmonary artery from those of vehicle-treated rats. These results suggest that the elevated muscular tone observed in the monocrotaline-induced hypertensive pulmonary artery is due to membrane depolarization of smooth muscle cells and that this phenomenon might be mediated by the activation of DIDS-sensitive Cl- channels. PMID- 11488432 TI - Involvement of GABAergic systems in manifestation of pharmacological activity of desipramine. AB - We have conducted this study to elucidate the influence of GABAergic systems on manifestation of pharmacological activity of desipramine using both pharmacological and electrophysiological methods. Desipramine (20 mg/kg, i.p.) significantly blocked the adjuvant-induced thermal hyperalgesia, which was facilitated by treatment with the GABA(A) antagonist picrotoxin (2 mg/kg, i.p.) or the GABA(B) antagonist saclofen (2 mg/kg, i.p.). This analgesic effect of desipramine was antagonized by post-treatment with picrotoxin or saclofen. However, none of these compounds showed any effect in normal animals without adjuvant-induced inflammation. In a slice preparation of the hippocampus, treatment with GABA (10(-5)-5 x 10(-4) M), baclofen (10(-5)-10(-4) M) or muscimol (10(-5)-10(-4) M) inhibited the field potential evoked in pyramidal neurons by Schaffer collateral stimulation. The inhibitory effect of GABA was facilitated by concurrent application of desipramine, carbamazepine or diazepam at a concentration of 5 x 10(-5)-2 x 10(-4) M. The rank of order of facilitation is: desipramine > carbamazepine > diazepam. Desipramine also enhanced the inhibitory effect of baclofen and muscimol. These results suggest that desipramine causes GABAergic systems to activate still more, and this phenomenon appears to be involved in manifestation of the pharmacological activity of desipramine such as antinociception. PMID- 11488433 TI - Hormonal effects of Z-350, possessing steroid 5alpha-reductase inhibitory and alpha1-adrenoceptor antagonistic actions, in the rat. AB - We examined the hormonal effects of Z-350, (S)-4-[3-(4-[1-(4-methylphenyl)-3-[4 (2-methoxyphenyl)piperazine-1-yl]propoxy]benzoyl)indole-1-yl]butyric acid hydrochloride, which has both alpha1-adrenoceptor blocking activity and steroid 5alpha-reductase inhibitory activity, in male and female rats. Z-350 administered orally for 14 days at a dose of 30 mg/kg to normal male rats significantly reduced the weight of the prostate and seminal vesicles without affecting the weight of the testis, epididymis, adrenals, kidney or liver. Prostatic levels of dihydrotestosterone decreased dose-dependently, with a slight increase in the level of testosterone at a Z-350 dose of 100 mg/kg. We observed no effects on the weight of the prostate in castrated rats or on the weight of the uterus in normal or 17beta-estradiol-treated female rats. These results suggest that Z-350 inhibits prostatic growth via inhibition of steroid 5-reductase without other hormonal effects. PMID- 11488434 TI - Protective effect of neurotropin against lipopolysaccharide-induced hypotension and lethality linked to suppression of inducible nitric oxide synthase induction. AB - Neurotropin is a non-protein extract from the dermis of rabbits inoculated with vaccinia virus and has been clinically used as an analgesic in Japan. We present in the current report evidence for its potential therapeutic value against endotoxin shock. Administration of this compound prior to lipopolysaccharide (LPS) challenge resulted in a reversal of a decrease of the mean arterial pressure in rats and also amelioration of lethality in mice. Anti-inducible nitric oxide synthase (iNOS) Western blotting of tissue extracts from LPS-treated mice revealed almost complete suppression of iNOS induction by Neurotropin. The findings in vivo were reproduced in in vitro experiments in which cultured human umbilical vascular endothelial cells were challenged with LPS. Simultaneous treatment of the cells with Neurotropin resulted in complete suppression of iNOS induction and significant reduction of cell death. These results suggested a therapeutic value of Neurotropin in the treatment of endotoxin shock that was linked, at least in part, to suppression of iNOS induction and reduced cell damage in vascular endothelial cells. PMID- 11488435 TI - Nociception and allodynia/hyperalgesia induced by intrathecal administration of fenvalerate. AB - The intrathecal injection of fenvalerate, a sodium channel activator, at doses of 0.01 to 3 microg, dose-dependently induced the duration of a characteristic behavioral syndrome mainly consisting of reciprocal hind limb scratching directed towards caudal parts of the body and biting or licking of the hind legs in mice. Fenvalerate-induced behavior was inhibited by morphine (1-10 mg/kg, i.p.). The characteristic behavior was also inhibited by mexiletine, a sodium channel blocker; MK-801, a N-methyl-D-aspartate ion-channel blocker; and GR82334, a neurokinin-1-receptor antagonist. Calphostin C (3 pmol, i.t.), a protein kinase C inhibitor, inhibited fenvalerate-induced behavior. On the other hand, phorbol-12, 13-dibutyrate (50 pmol, i.t.), a protein kinase C activator, markedly enhanced the fenvalerate-induced behavior. The present results also showed that fenvalerate produced thermal allodynia and hyperalgesia in the tail-flick test. Furthermore, fenvalerate-induced thermal allodynia and hyperalgesia were inhibited by the pretreatment with calphostin C. These results suggest that the intrathecal administration of fenvalerate induces a marked nociceptive response and thermal allodynia/hyperalgesia, and they suggest that tetrodotoxin-resistant sodium channels may play an important role in this effect. PMID- 11488436 TI - Usefulness and limitation of DiBAC4(3), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials regulated by recombinant large conductance Ca2+-activated K+ channels in HEK293 cells. AB - The usefulness of bis-(1,3-dibutylbarbituric acid)-trimethine oxonol (DiBAC4(3)), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials (MPs) was evaluated in HEK293 cells, where alpha or alpha plus beta1 subunits of large conductance Ca2+-activated K+ (BK) channels were expressed (HEKBK alpha and HEKBK alphabeta). The fluorescent intensity of DiBAC4(3) was measured at various potentials under voltage-clamp for calibration to estimate the absolute MP semi quantitatively. The resting MPs measured with DiBAC4(3) were roughly comparable to those recorded with a microelectrode; the MP in HEKBK alphabeta was 10-20 mV more negative than that in native HEK. In HEKBK alpha, the membrane hyperpolarization induced by 10 microM Evans blue, a BK channel opener, was detected with DiBAC4(3). NS-1619, another BK channel opener, induced gradual but substantial change in F/F(K) even in native HEK, while the BK channel opening effect was detected. Oscillatory membrane hyperpolarization was induced in HEKBK alphabeta by application of 10 microM acetylcholine via increase in intracellular Ca2+ concentration. The oscillatory hyperpolarization was, however, detected only as a slow hyperpolarization with DiBAC4(3). It can be concluded that relatively slow effects of BK channel modulators can be semi-quantitatively measured by use of DiBAC4(3) in HEKBK, while the limited temporal resolution and possible artifacts should be taken into account. PMID- 11488437 TI - Transient prevention of ethanol-induced gastric lesion by capsaicin due to release of endogenous calcitonin gene-related peptide in rats. AB - Pre-exposure of the rat gastric mucosa to capsaicin reduced the mucosal lesion by 50% ethanol to 1/4. Treatment with an antagonist of calcitonin gene-related peptide (CGRP), CGRP (8-37), nullified the effect of capsaicin. During constant perfusion of the gastric lumen with physiological saline + pepstatin, the CGRP level was not increased by 50% ethanol, but it showed a peak (802.5 +/- 145.7 pg/2 min) after 1.6 mM capsaicin. Four minutes after capsaicin, the CGRP level was kept at a high level and the gastric lesion was markedly reduced by re exposure of the mucosa to 50% ethanol. At 20-30 min after capsaicin, the CGRP levels returned to the resting level and the reddened area by 50% ethanol was not reduced. It was concluded that capsaicin transiently prevented the mucosal lesion through CGRP release. PMID- 11488438 TI - Time-dependent apoptotic development and pro-apoptotic genes expression in rat heart after myocardial infarction. AB - We investigated the apoptotic development and apoptotic-related gene expression after myocardial infarction (MI) at different time points in the current study. Bax gene expression was increased at 12 h after MI and peaked at 24 h. Fas gene started to over-express at 12 h after MI as well but it reached maximum at 72 h. In the MI groups, strongest staining of apoptosis was detected in rats 3 days post operation. Our results demonstrate that apoptotic development after MI is time dependent in the ischemic area and there could be some linkage with the over expression of angiotensin II receptors post MI. PMID- 11488439 TI - Effects of HSP-117, a novel tachykinin NK1-receptor antagonist, on cisplatin induced pica as a new evaluation of delayed emesis in rats. AB - The effects of a novel tachykinin NK1-receptor antagonist HSP-117 [(2S,3S)-3-[(5 isopropyl-2,3-dihydrobenzofuran-7-yl)methyl]amino-2-phenylpiperidine dihydrochloride] on cisplatin-induced pica, i.e., the eating of nonnutritive substances such as kaolin were examined in rats. HSP-117 inhibited kaolin intake in a dose-dependent manner for 2 days. The 5-HT3-receptor antagonist ondansetron inhibited only on the first day, but not on the second day. These results indicate that the cisplatin-induced kaolin intake on the first day is related to both 5-HT3- and NK1 receptors, while only the NK1 receptor is involved on the second day. Thus, cisplatin-induced continuous pica in rats represents a useful model of not only acute but also delayed emesis. PMID- 11488440 TI - Effects of beta-casomorphins on neuronal survival in culture of embryonic chick dorsal root ganglion neurons. AB - We studied the effects of beta-casomorphins (beta-CMs, mu-acting opioid peptides from milk protein beta-casein) on survival of primary-cultured chick dorsal root ganglion neurons in the presence of nerve growth factor. Beta-CM-5 and beta-CM-7 had potent neuronal survival-promoting activities. Beta-CM-4 amide (morphiceptin) and des-Tyr'-beta-CM-7 also exhibited the similar promoting effects, although their effects were very weak. The promoting effect of beta-CM-5 was prevented by co-administration of naloxone, or pretreatment with pertussis toxin. These results suggest that the neuronal survival-promoting effects of beta-CMs might be mediated through opioid receptors coupled to G proteins. PMID- 11488441 TI - Opposite effects of calcium and magnesium on the central blood pressure regulation in the spontaneously hypertensive rats. AB - The effects of intracerebroventricular administration of calcium or magnesium on the blood pressure regulation in the brain were investigated. The systolic blood pressure in spontaneously hypertensive rats (male, 13-week-old) was decreased by calcium chloride (100 microg/rat) and increased by magnesium chloride (20, 100 or 500 microg/rat). The depressor response induced by calcium was inhibited by magnesium chloride in a dose-dependent manner. Combining these results with those previously reported, it is suggested that magnesium inhibits the ability of calcium to reduce blood pressure through calmodulin- and dopamine-dependent functions in the brain. PMID- 11488442 TI - Lessons learned. PMID- 11488443 TI - Malaria control through impregnated bednets--a pilot project in selected villages in Lao PDR. AB - In 7 villages on the foothills of the Houayxai district of the Bokeo Province in Lao PDR between midyear 1995 to midyear 1997 an attempt was made to test the acceptability and use of DDT impregnated bed nets as well as environmental and behavioral risk factors. About 380 women between 15 to 45 years old and their children in the age range of 1 to 14 years had been studied. A pre-tested questionnaire had been applied and blood from women and children was taken from the finger prick and a conventional thick and thin blood smear was performed, fixed Giemsa stained and examined for malaria parasites. DDT (dichlorodiphenyltrichloroethane) impregnated mosquito nets were distributed in the intervention villages. The availability of mosquito nets increased statistically significant from approximately 50% to 70% for all family members in the intervention area between 1995 and 1997 and statistically significant decreased in the control area from 79% to 45.1%. There was a statistically significant decrease in malaria attacks as claimed by the females for the intervention area but not for the control villages. The proportion of positive blood smears did decrease overall for women and children in between 1995 to 1997. Occupation, location of the house and use of mosquito nets had been determined as the most important risk factors. PMID- 11488444 TI - Community based intervention of iron deficiency anemia in females and iodine deficiency disorders in school children in Lao PDR. AB - Under the supervision of the central and local health authorities, a pilot project was conducted in four villages in the Luangprabang Province, Lao PDR. The objective of the project was to test different regimes to supplement females with oral iron preparations to reduce iron deficient anemia (IDA) and control iodine deficiency disorders (IDD) in school children. Compared with iron sulphate tablets, iron fumerate tablets were well accepted and good compliance results were achieved. Hemoglobin concentration improved only in the group of females taking iron fumerate tablets. The goiter rate decreased from approximately 90% to about 45% for school children, regardless of whether iodine salt were used by their families or whether iodine capsules were used to treat the children. The latter attempt was hampered by the fact that also in the control village iodine fortified salt was used. This was due to a governmental attempt to control IDD nation-wide. Therefore, also in the control village a significant decrease in the goiter rate was observed. PMID- 11488445 TI - Control of intestinal parasitic infection--a pilot project in Lao PDR. AB - Intestinal helminths are a very common but still rather unrecognized public health problem. An attempt was undertaken to control the three important parasites, ie Ascaris, Trichuris and hookworm. Two villages in Vientiane Province in Lao PDR were selected, one as an intervention and the other as control village. Intervention measures started by providing mass treatment against Ascaris, Trichuris, hookworm and other intestinal parasitic infections in both villages. Health education and other intervention measures were implemented in the intervention village, which significantly influenced the re-infection rates in this village. These achievements were not due to an improvement of the availability of toilets or personal hygiene alone but more to the villagers' improved understanding of the route of the parasitic infections. It was concluded that intervention methods should be directed more towards particular age groups. Particular attention should be paid to control parasitic infections among females. PMID- 11488446 TI - Promotion of the health of rural women towards safe motherhood--an intervention project in northeast Thailand. AB - An intervention project focusing on the health of women in the reproductive age was conducted in three districts of Khon Kaen Province, northeast Thailand between 1991 and 1996. Main emphasis was placed on improving reproductive health, the nutritional status including the iron deficiency anemia (IDA) as well as iodine deficiency disorders (IDD), and the parasitic diseases liver fluke (Opisthorchis viverrini) and hookworm. For implementation a community based Primary Health Care approach was used including the training of health officials in health matters, primary health care workers and villagers as well as enhancing health education and the dissemination of health information. The health delivery system was encouraged to take appropriate actions such as in the treatment of parasitic diseases and the control of IDA and IDD. Monitoring was done on a regular basis. The outcome of the project was assessed by comparing baseline data compiled from a random sample of the target population with the results of the final evaluation. An attempt to compare results obtained from villages within and outside of the project area failed most probably because of spill over effects. A number of important indicators on family planning and mother and child health care improved during the time the project was implemented; this included practising family planning, and participation in antenatal care. Also the proportion of females becoming pregnant for the first time when 20 years or older increased. Child-raising also improved in that almost all females gave colostrum to their babies by this time. Almost 75% of the women breast-fed their children. Improvements occurred in the nutritional status as far as the micronutrients iron and iodine were concerned, however the overall nutritional status of females did not change, but a rather high proportion of females were found to be overnourished. The project failed in reducing abortion and the proportion of females becoming pregnant when they are 18 years old or younger. It was also not possible to improve the usage of postnatal care. As anticipated, the results achieved so far are most suitable in serving as a training ground and providing a favorable example to improve family planning, mother- and child health care, and also the general health of females in the region, particularly in neighboring countries such as Lao PDR, Cambodia and Vietnam. PMID- 11488447 TI - Reproductive health in selected villages in Lao PDR. AB - The aim of the project was to improve the knowledge and attitude towards birth spacing by training the villagers in the selected villages of Vientiane Province in Lao PDR in family planning, providing them with the various family planning methods, and improving antenatal (ANC) and postnatal (PNC) care in the villages. Throughout the province, traditional birth attendants (TBA) were trained on several occasions during the project period. There were clear indications that reproductive health improved between 1995 and 1997. Considerable improvements were observed in the percentage of women making use of ANC and practising birth spacing by using some form of contraception or other. The most common methods used were contraceptive pills and injectables. In the case of child mortality a slight decrease was found in the percentage of women having their first pregnancy below the age of 18 years. A still unsolved problem is the high number of abortions. PMID- 11488448 TI - Nutritional status of pre-school children and women in selected villages in the Suvannakhet Province, Lao PDR--an intervention trial. AB - A project to promote the health and nutritional status of women and pre-school children was started from 1995 to 1997 in three villages in Suvannakhet Province, Lao PDR. One village served as control. In 1995, for the baseline survey, 456 females, and in 1997, for the final evaluation, 363 females from the four villages volunteered for further investigations. An attempt was made to involve all females in the reproductive age residing in the villages. At the same time also the nutritional status of 321 and about 540 randomly selected pre-school children respectively was also assessed through physical examination and anthropometric measurements. Intervention measures included introducing growth charts and taking regular anthropometric measurements of women in the reproductive age and of pre-school children. Training in nutritional aspects such as giving colostrum to new-borns, prepare proper weaning food and supplementary feeding, animal-raising and home gardening was also introduced and provided to health personnel, village leaders and in women clubs. Special attention was given to the control of acute infectious diseases. The conventional EPI program was enforced as well. Health education in matters of mother and child health care was also provided. The proportion of undernourished women was rather high at about 15%. For pre-school children, the proportion of wasting was around 5%, and of stunting 50% and above. Intervention did not improve the nutritional status either of the women or of the children. It was concluded that the time span of two years is too short for a decrease in the proportion of undernourishment to be observed. An improvement was achieved for some indicators of mother and child health care. This seems to indicate the population's willingness to follow suggestions to improve their health. Most probably, if attempts to improve the nutritional status were continued, an improvement in this aspect could also be observed, if the population can be encouraged to take actions and develop initiatives by themselves. PMID- 11488449 TI - Effects of gum resin of Boswellia serrata in patients with chronic colitis. AB - Patients studied here suffered from chronic colitis characterized by vague lower abdominal pain, bleeding per rectum with diarrhoea and palpable tender descending and sigmoid colon. The inflammatory process in colitis is associated with increased formation of leukotrienes causing chemotaxis, chemokinesis, synthesis of superoxide radicals and release of lysosomal enzymes by phagocytes. The key enzyme for leukotriene biosynthesis is 5-lipoxygenase. Boswellic acids were found to be non-redox, non-competitive specific inhibitors of the enzyme 5 lipoxygenase. We studied the gum resin of Boswellia serrata for the treatment of this disease. Thirty patients, 17 males and 13 females in the age range of 18 to 48 years with chronic colitis were included in this study. Twenty patients were given a preparation of the gum resin of Boswellia serrata (900 mg daily divided in three doses for 6 weeks) and ten patients were given sulfasalazine (3 gm daily divided in three doses for 6 weeks) and served as controls. Out of 20 patients treated with Boswellia gum resin 18 patients showed an improvement in one or more of the parameters: including stool properties, histopathology as well as scanning electron microscopy, besides haemoglobin, serum iron, calcium, phosphorus, proteins, total leukocytes and eosinophils. In the control group 6 out of 10 patients showed similar results with the same parameters. Out of 20 patients treated with Boswellia gum resin 14 went into remission while in case of sulfasalazine remission rate was 4 out of 10. In conclusion, this study shows that a gum resin preparation from Boswellia serrata could be effective in the treatment of chronic colitis with minimal side effects. PMID- 11488450 TI - Effect of an extract of the root of Scutellaria baicalensis and its flavonoids on aflatoxin B1 oxidizing cytochrome P450 enzymes. AB - The inhibition of aflatoxin B1 (AFB1) metabolism by a water extract of the root of Scutellaria baicalensis and its flavonoids was examined in liver microsomes. AFB1 is known to be metabolized to aflatoxin M1 (AFM1), aflatoxin Q1 (AFQ1), and AFB1-8,9-epoxide (AFBO). The water extract potently inhibited the production of AFM1 by cytochrome P450 (CYP)1A1/2 and slightly reduced AFBO formation by CYP1A1/2, CYP2B1, CYP2C11 and CYP3A1/2 in TCDD-treated rat liver microsomes. IC50 values for AFM1 and AFBO formation were 6.8 and 122.4 microg/ml, respectively. Wogonin showed the highest inhibitory activity towards AFM1 formation among the flavonoids isolated from the extract. On the other hand, the extract had no effects on the formation of AFBO and AFQ1 in human liver microsomes, and on the activities of CYP2B1, CYP2C11 and CYP3A1/2 which were detected by hydroxylation patterns of testosterone. These results demonstrated that the extract of the root of Scutellaria baicalensis has a specific inhibitory effect on CYP1A1/2 among CYP enzymes involved in AFB1 metabolism by rat and human microsomes. PMID- 11488451 TI - Expression of protein kinase C isoforms in euxanthone-induced differentiation of neuroblastoma cells. AB - Euxanthone, a potent neuritogenic compound isolated from the roots of the medicinal herb Polygala caudata, has recently been shown to induce the differentiation of murine neuroblastoma Neuro 2A (BU-1) cells. In this study, the role of protein kinase C (PKC) and the expression of various PKC isoforms in euxanthone-treated BU-1 cells were examined. mRNA phenotyping using the reverse transcription polymerase chain reaction (RT-PCR) showed that BU-1 cells express six different PKC isoforms, namely PKC-alpha, -beta, -delta, -epsilon, -lambda, and -zeta. Differential regulation and expression of PKC isoforms was observed in BU-1 cells treated with 100 microM euxanthone. PKC-apha, -beta, -delta, -lambda and -zeta were all up-regulated, with 1.7- to 9.5-fold increase, at around 30 to 60 minutes after euxanthone treatment. The expression level of PKC-epsilon remained relatively constant during the treatment. PKC-gamma, -eta, and -theta were not detected in both untreated and euxanthone-treated BU-1 cells. Staurosporine, a broad spectrum PKC inhibitor, was found to inhibit both spontaneous and euxanthone-induced neuritogenesis in BU-1 cells. A significant reduction of the euxanthone-induced neuritogenic effect was also observed when the PKC isoform-specific inhibitor Go6976 was included in the culture. These results suggest that the euxanthone-induced differentiation of the neuroblastoma BU-1 cells may be mediated through the differential expression of PKC-alpha, beta, -delta, -lambda and -zeta isoforms. PMID- 11488452 TI - Kaurane diterpenes from Isodon japonicus inhibit nitric oxide and prostaglandin E2 production and NF-kappaB activation in LPS-stimulated macrophage RAW264.7 cells. AB - A methanolic extract of the whole plant of Isodon japonicus (Labiatae) showed potent inhibition on the LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production in RAW264.7 cells. Four known kaurane diterpenes were isolated by activity-guided fractionation and their structures were identified as kamebanin (1), kamebacetal A (2), kamebakaurin (3), excisanin A (4). All compounds also inhibited the LPS-induced NF-kappaB activation as assessed by NF kappaB reporter assay and electrophoretic mobility shift assay (EMSA). Compounds 2-4 showed comparable inhibitory effects on the LPS-induced production of NO and PGE2, and activation of NF-kappaB without affecting cell viability. These results suggest that kaurane diterpenes could exert their inhibitory effects on the production of NO and PGE2 through the suppression of NF-kappaB activation, and be partially responsible for the anti-inflammatory activities of the genus Isodon. PMID- 11488453 TI - Identification and isolation of the cyclooxygenase-2 inhibitory principle in Isatis tinctoria. AB - Various extracts prepared from the traditional dye and medicinal plant Isatis tinctoria L. were submitted to a broad in vitro screening against 16 anti inflammatory targets. Dichloromethane (DCM) extracts from dried leaves showed a marked cyclooxygenase (COX) inhibitory activity with a preferential effect on COX 2 catalysed prostaglandin synthesis. A supercritical fluid extraction (SFE) procedure employing CO2-modifier mixtures was developed by which the bioactivity profile and chromatographic fingerprint of the DCM extract could be reproduced. High-resolution activity directed on-line identification of the COX-2 inhibitory principle, using a combination of LC-DAD-MS with a microtitre-based bioassay, led to the identification of tryptanthrin (1) as the constituent responsible for essentially all COX-2 inhibitory activity in the crude extract. Following on-line identification, 1 was isolated at preparative scale and its structure confirmed by comparison with synthetic tryptanthrin. In an assay with lipopolysaccharide stimulated Mono Mac 6 cells, tryptanthrin (1) was of comparable potency (IC50 = 64 nM) than the preferential COX-2 inhibitors nimesulide (IC50 = 39 nM) and NS 398 (IC50 = 2 nM). The SFE extract and 1 showed no cytotoxicity in Mono Mac 6 and RAW 264.7 cells when tested at 100 microg/ml and 10 microM, respectively. PMID- 11488454 TI - Effects of ginsenosides from Panax ginseng on cell-to-cell communication function mediated by gap junctions. AB - Gap junctions have been shown or are believed to be involved in the pathogenesis of many inherited and acquired human diseases. Agents that regulate the gap junction-mediated intercellular communication (GJIC) function may facilitate prevention and treatment of GJIC-involved diseases. In the present study we examined the effects of 27 ginsenosides isolated from Panax ginseng on GJIC. The results show that compounds 1 (oleanolic acid), 2 (ginsenoside-R0), 3 (ginsenoside-Rb1), 5 (ginsenoside-Rb2), 7 (ginsenoside-Rd), 8 (ginsenoside-Rg3), 12 (panaxadial), 13 (notoginsenoside-R4), 17 [ginsenoside-Rg2 (20S)], 18 (ginsenoside-Rf), and 26 (ginsenoside-F3) did not obviously affect GJIC, whereas compounds 4 (ginsenoside-Rc), 6 (ginsenoside-Rb3), 9 (ginsenoside-Rd2), 10 (notoginsenoside-Fe), 11 (ginsenoside-Rh2),14 (ginsenoside-Ra1), 15 (ginsenoside Re), 16 [ginsenoside-Rg2 (20R)], 19 (ginsenoside-Ia), 20 [ginsenoside-Rh1 (20S)], 21 [ginsenoside-Rh1 (20R)], 22 (ginsenoside-F1), 23 (protopanaxatriol), 24 (panaxatriol), 25 (ginsenoside-Rg1), and 27 (chikusetsaponin-L8) induced GJIC reductions at various degrees. Compounds 2, 7, and 8 protected against the tyrosine phosphatase inhibitor vanadate-induced GJIC reduction, while compounds 1, 5, 7, and 17 inhibited the cytokine interleukin 1 alpha (IL-1alpha)-induced reduction in GJIC. Nevertheless, no compounds protected against the protein kinase C (PKC) activator 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced GJIC inhibition. On the other hand, GJIC reductions induced by compounds 6, 9,10, 20, 21, 22, 24, and 25 were inhibited by the tyrosine kinase (TK) inhibitor genistein, while GJIC reductions induced by compounds 6, 9, 14, 16, 19, 21, and 24 were attenuated in the presence of the PKC inhibitor calphostin C. However, GJIC reductions induced by compounds 4, 23, and 27 were not inhibited either by genistein or by calphostin C. These data indicate that various mechanisms are responsible for effects of ginsenosides on GJIC. PMID- 11488455 TI - A new tetrahydroprotoberberine N-oxide alkaloid and anti-platelet aggregation constituents of Corydalis tashiroi. AB - A new tetrahydroprotoberberine N-oxide alkaloid, (-)-cis-isocorypalmine N-oxide (1), together with two known compounds, 6-methoxydihydrosanguinarine (2) and norjuziphine (3), were isolated in continuing studies of the entire Formosan Corydalis tashiroi plant. The structures of these three compounds were determined through spectral analyses. In addition, compounds 1, 2, 3 and the seven alkaloids previously reported: (-)-cis-corydalmine N-oxide, (-)-trans-corydalmine N-oxide, (-)-trans-isocorypalmine N-oxide, scoulerine, protopine, oxysanguinarine and corydalmine, were found to possess antiplatelet aggregation activity. PMID- 11488456 TI - Acidic polysaccharides from rhizomes of Atractylodes lancea as protective principle in Candida-lnfected mice. AB - Prophylactic effects upon imunnosuppressed mice lethally infected by Candida albicans were examined in fractions prepared from a constituent herb of Juzen taiho-to (TJ-48, Si-Quan-Da-Bu-Tang), rhizomes of Atractylodes lancea DC. The oral administration of water extract obtained from a residue after MeOH extraction of rhizomes significantly prolonged the survival period of the infected mice at a dose of 140 mg/kg/day compared with control mice, while the MeOH extract did not. In the crude polysaccharide fraction (F-2) obtained by EtOH precipitation of the water extract, a significant life-prolonging effect was observed by the administration of 70 mg/kg/day. F-2 was further fractionated, and the resulting strongly acidic polysaccharide fraction, F-2-2, had a protective effect at a dose of 17.5 mg/kg/day. This fraction mainly consisted of acidic pectic polysaccharides containing about 80% galacturonic acid. The protective activity of F-2-2 was lost by periodate oxidation, but not by protease digestion, suggesting that the polysaccharide component of F-2-2 plays a major role in the protective activity against Candida-infected mice. PMID- 11488457 TI - Production of withaferin A in shoot cultures of Withania somnifera. AB - Multiple shoot cultures of Withania somnifera were established from single shoot tip explants and their potential for the production of two principle withanolides, withaferin A and withanolide D was investigated. Shoot tips grown on MS medium supplemented with BA (1 mg l(-1)) induced 10.0 +/- 1.15 microshoots per explants and shoot cultures accumulated both withanolides (withaferin A = 0.04%, withanolide D = 0.06%). Supplementation of MSSM (solid) agar medium with 4% sucrose enhanced accumulation of both withaferin A (0.16%) and withanolide D (0.08%). Reduction of the agar concentration to 0.16% increased the number of microshoots induced per explant to 25.5. MSSM liquid medium containing 10% coconut milk favoured a maximum increase in biomass (27 fold); number of microshoots induced (37.6 +/- 1.45) as well as accumulation of withaferin A (0.14%). PMID- 11488458 TI - Further phenols and polyacetylenes from the rhizomes of Atractylodes lancea and their anti-inflammatory activity. AB - From the rhizomes of Atractylodes lancea, 2-[(2'E)-3',7'-dimethyl-2',6' octadienyl]-4-methoxy-6-methylphenol (1) was isolated as a new natural product. The compound showed strong inhibitory effects on 5-lipoxygenase (5-LOX) and cyclooxygenase-1 (COX-1), but exhibited only weak antioxidative activities [IC50 = 0.1 microM (5-LOX), 2 microM (COX-1), 9 microM (PMN/FMLP), 28 microM (PMNIOZ)]. Moreover, five new acetylenes were isolated and elucidated as (3Z,5E,11E) tridecatriene-7,9-diynyl-1-O-(E)-ferulate (2), erythro-(1,3Z,11E)-tridecatriene 7,9-diyne-5,6-diyl diacetate (3), (1Z)-atractylodin (4), (1Z)-atractylodinol (5), (1Z)-acetylatractylodinol (6) plus the known (4E,6E,12E)-tetradecatriene-8,10 diyne-1,3-diyl diacetate (7). Among the acetylenes, only 2 showed strong inhibition of 5-LOX and COX-1 activity (IC50 (5-LOX) = 3 microM, IC50 (COX-1) = 1 microM). In addition, the fatty acids linoleic acid, oleic acid and palmitic acid with previously established 5-LOX-/COX-1 inhibitory actions were identified as major constituents of the n-hexane extract and thus seem to contribute to the plant's in vitro activity. PMID- 11488459 TI - Cytotoxic activity of moronic acid and identification of the new triterpene 3,4 seco-olean-18-ene-3,28-dioic acid from Phoradendron reichenbachianum. AB - The cytotoxic compound moronic acid (1) and the new tetracyclic triterpene 3,4 seco-olean-18-ene-3,28-dioic acid (2), were isolated from the aerial parts of the medicinal plant Phoradendron reichenbachianum (mistletoe, Loranthaceae) through a bioassay-guided fractionation. In addition, squalene, glycerol trilinoleate, morolic acid, betulonaldehyde, betulinaldehyde, alpha-germanicol, lupeol, beta sitosterol and beta-sitosteril glucopyranoside, were identified in this plant species. The structures were elucidated on the basis of chemical and spectroscopic evidence. PMID- 11488460 TI - Improved method for the determination of oxindole alkaloids in Uncaria tomentosa by high performance liquid chromatography. AB - This Paper describes an improved HPLC method for the determination of pentacyclic oxindole alkaloids in Uncaria tomentosa (Cat's Claw). Six of the isomeric compounds could be baseline separated at room temperature within less than 30 min by using 3 microm C-18 column material and a mobile phase consisting of 10 mM phosphate buffer at pH 7.0 and acetonitrile. At a wavelength of 245 nm all standard compounds could be detected at concentrations as low as 0.63 microg/ml. Different samples of U. tomentosa bark and market products containing Cat's Claw were extracted with a modified procedure ensuring the integrity of the alkaloids and analyzed successfully. The results indicated accuracy and consistency of the new method, and showed variations in the total alkaloid content in products from 0.156 to 0.962%. PMID- 11488461 TI - Supercritical fluid extraction and analysis of compounds from Clivia miniata for uterotonic activity. AB - In this descriptive study, the superciritical fluid extract of the roots of Clivia miniata L. was tested for uterotonic activity using guinea pig uterine smooth muscle in vitro. Extraction was performed with water modified supercritical carbon dioxide at 400 atm and 80 degrees C. The uterine contractions induced by this extract were compared to those induced by the aqueous extract and found to be active at lower doses. The active compounds were isolated and the structures elucidated by spectroscopic and chromatographic techniques. Both linoleic acid and 5-hydroxymethyl-2-furancarboxaldehyde isolated from the extract were found to induce muscle contractions individually. The pharmacological mode of action of 5-hydroxymethyl-2-furancarboxaldehyde was assessed using two receptor agonists and antagonists. This compound was found to mediate its effect through cholinergic receptors. PMID- 11488462 TI - Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA. AB - Herba Dendrobii (Shihu) is a commonly used Chinese medicine derived from the stem of several orchid species belonging to the genus Dendrobium. It is rather expensive and adulteration is frequent. Proper authentication of the medicinal species is necessary to protect consumers and support conservation measures. DNA sequences of the internal transcribed spacer 2 (ITS 2) of 16 Dendrobium species were shown to be significantly different from one another by an average of 12.4% and from non-orchids and Pholidota (an adulterant of Shihu) by 29.8% and 18.8%, respectively. The intra-specific variation among the Dendrobium species studied was only about 1%. Therefore, ITS 2 regions could be adopted as a molecular marker for differentiating medicinal Dendrobium species from one another and also from non-orchids and adulterants. PMID- 11488463 TI - Phylogenetic analysis based on 18S rRNA gene and matK gene sequences of Panax vietnamensis and five related species. AB - Panax vietnamensis was discovered recently in Vietnam. Its bamboo-like rhizomes, called Vietnamese Ginseng, have attracted considerable attention because of their specific pharmacological activities. In order to define the taxonomic position of this new species and include it in the molecular authentication of Ginseng drugs, the 18S ribosomal RNA gene and matK gene sequences of P. vietnamensis were determined and compared with those of its related taxa, P. japonicus var. major and P. pseudo-ginseng subsp. himalaicus, besides previously reported P. ginseng, P. japonicus and P. quinquefolius. The 18S rRNA gene sequences were found to be 1809 bps in length. The sequence of P. vietnamensis was identical to that of P. quinquefolius, and presented one base substitution from those of both P. japonicus var. major and P. pseudo-ginseng subsp. himalaicus. The matK gene sequences of 6 taxa were found to be 1509 bps in length. The sequence of P. vietnamensis differed from those of P. japonicus var. major, P. pseudo-ginseng subsp. himalaicus, P. ginseng, P. japonicus and P. quinquefolius at 4, 5, 9, 9 and 10 nucleotide positions, respectively. The phylogenetic tree reconstructed by the combined 18S rRNA-matK gene analysis using the maximum parsimony method showed that P. vietnamensis was sympatric with other Panax species and had a close relationship with P. japonicus var. major and P. pseudo-ginseng subsp. himalaicus. PMID- 11488464 TI - Inhibition of platelet-activating factor receptor binding by aporphine and phenanthrenoid alkaloids from Aromadendron elegans. AB - Six aporphine and one phenanthrenoid alkaloids isolated from Aromadendron elegans Blume were investigated for their inhibitory effect on platelet-activating factor (PAF) binding to rabbit platelets using 3H-PAF as a ligand. Of the compounds tested, (-)-N-acetylanonaine, 1-(N-acetyl-N-methylamino)ethyl-3,4,6-trimethoxy-7 hydroxy-phenanthrene, and predicentrine showed strong inhibition of PMID- 11488465 TI - Flavonoids as cycline-dependent kinase inhibitors: inhibition of cdc 25 phosphatase activity by flavonoids belonging to the quercetin and kaempferol series. AB - In an effort to detect potential inhibitors of cdc25 phosphatase, nineteen flavonoids belonging to the quercetin and kaempferol series have been evaluated, using a colorimetric assay of recombinant human cdc25A tyrosine phosphatase as a cell cycle-specific target. Compounds bearing two benzyl or methyl groups in positions 7 and 4' and acetyl on the hydroxy groups of the sugar moiety showed the maximal activity. PMID- 11488466 TI - Neuroprotective dibenzylbutyrolactone lignans of Torreya nucifera. AB - The methanolic extract of the bark of Torreya nucifera Sieb. et Zucc. (Taxaceae) significantly protected primary cultures of rat cortical cells exposed to the excitotoxic amino acid, L-glutamate. (-)-Arctigenin (1), (-)-traxillagenin (2), arctiin (4), traxillaside (5), and a newly-reported compound 3 (-)-4' demethyltraxillagenin [(2R,3R)-2-(4''-hydroxy-3''-methoxybenzyl)-3-(4'-hydroxy 3',5'-dimethoxybenzyl)-butyrolactone] were isolated by bioactivity-guided fractionation and further separation using chromatographic techniques. These lignans and their glycosides had significant neuroprotective activities against glutamate-induced toxicity in primary cultures of rat cortical cells at concentrations ranging from 0.01 microM to 10.0 microM. PMID- 11488467 TI - Chemopreventive activity of isoquinoline alkaloids from Corydalis plants. AB - Eighteen isoquinoline alkaloids including protoberberines (1-12), benzophenanthridines (13-16) and an aporphine (17) isolated from plants of Corydalis species (Fumariaceae) were tested for inhibitory effects on Epstein Barr virus early antigen activation induced by 12-O-tetradecanoylphorbol 13 acetate in Raji cells. In a primary screening test, all of the isoquinoline alkaloids showed inhibitory activity with the IC50 values being in the range of 140-410 mol ratio/32 pmol TPA. The data demonstrate that these isoquinoline alkaloids might be valuable as anti-tumor promoters. PMID- 11488468 TI - Synthesis and biosynthesis of isocordoin. AB - In the search of a convenient synthesis for isocordoin (1), a potential anticancer natural product, 2',4'-dihydroxychalcone was inoculated in cell suspension cultures of Morus nigra, which were expected to contain an active prenyltransferase. After 24 hours the target compound was easily isolated from the metabolite extract. Optimization of the biotransformation resulted in a 85% yield of the prenyl derivative. PMID- 11488469 TI - Cardanols from leaves of Rhus thyrsiflora. AB - A mixture of 3-substituted alkyl- and alkenylphenols including nine new compounds (cardanols) was isolated from leaves of the Yemenian plant Rhus thyrsiflora (Anacardiaceae) and identified by GC-MS. The position of the double bond in the compounds bearing a monolefinic side chain was determined by their typical MS fragmentation patterns after hydroxylation and trimethylsilylation. PMID- 11488470 TI - Absolute configuration of (6S,9S)-roseoside from Polygonum hydropiper. AB - From Polygonum hydropiper L., a C13-norisoprenoid glucoside was isolated and its absolute configuration was established to be (6S,9S)-roseoside (1) by spectroscopic evidence and X-ray crystallographic analysis of its acetate derivative (2). In addition, the stereostructure of roseoside from Canthium subcordatum was revised to the (6S,9S) configuration. PMID- 11488471 TI - An acetylene and a monoterpene glycoside from Adenocaulon himalaicum. AB - The methanolic extract of the aerial parts of Adenocaulon himalaicum (Asteraceae) has yielded a new acetylene, 1-O-feruloyl-tetradeca-4E,6E,12E-triene-8,10-diyne (1), a new monoterpene glycoside, 9-hydroxylinaloyl-3-O-(4-O-coumaroyl)-beta-D glucopyranoside (2), and eight known compounds. Their structures were established by chemical and spectroscopic methods. PMID- 11488472 TI - Triterpenoid saponins from Clematis tangutica. AB - Two new triterpenoid saponins, tanguticoside A and B along with seven known saponins vitalboside B, alpha-hederin, saponin PK, beta-hederin, saponin PJ3, saponin PE, and ciwujianoside A were isolated from aerial part of Clematis tangutica. By chemical and spectral evidences methods, the structures of tanguticoside A and B were elucidated as 3-O-beta-D-glucopyranosylhederagenin 28 O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranoside and 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D glucopyranosylhederagenin 28-O-alpha-D-rhamnopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranoside, respectively. PMID- 11488473 TI - Quantitative regulation of the Flaveria Me1 gene is controlled by the 3' untranslated region and sequences near the amino terminus. AB - The Me1 gene of Flaveria bidentis codes for the C4 isoform of NADP malic enzyme, which accumulates to a high-level only in bundle sheath cells. Previous experiments demonstrated that sequences at the 5' end of the gene control cell specificity whereas sequences at the 3' end are necessary for high-level expression. To localize quantitative regulator sequences, we have analysed a series of Me1 3' deletion constructs fused to the gusA reporter gene. We show that sequences within the 3'-untranslated region (3'-UTR) control quantitative levels of expression. Analysis of 5' promoter fusions demonstrated that high level expression also requires sequences within the N-terminal coding region of the gene, suggesting possible interactions between the 3'-UTR and 5' coding regions. Cell-specific regulatory sequences are located in a different part of the 5' end of the gene, between 1023 bp upstream of the transcription start and the start of translation. PMID- 11488474 TI - Two Arabidopsis thaliana genes, KOR2 and KOR3, which encode membrane-anchored endo-1,4-beta-D-glucanases, are differentially expressed in developing leaf trichomes and their support cells. AB - The Arabidopsis KOR gene encodes a membrane-anchored endo-1,4-beta-D-glucanase involved in cell wall assembly. To obtain a more detailed knowledge of the small gene family encoding membrane-anchored endo-1,4-beta-D-glucanases in Arabidopsis thaliana, we have characterized two additional membrane-anchored endo-1,4-beta-D glucanase genes. Sequence comparison indicates that KOR2 is distantly related to KOR and other plant membrane-anchored endo-1,4-beta-D-glucanases. The expression of KOR2 and KOR3 was followed by the beta-glucuronidase (gusA) reporter-gene method. While the KOR gene is most often expressed throughout the plant, KOR2::gusA and KOR3::gusA are active only in restricted cell types. We demonstrate that KOR2::gusA is expressed very early in the development of root hairs within the root differentiation zone (specialization zone) but not in the root-hair-bearing epidermal cells at the root/shoot junction (transition zone). Furthermore, KOR2::gusA is expressed in the proximal parts of leaves and floral organs (rosette and cauline leaves, sepals, petals and stamens), and in trichomes, as they develop at the tip of young leaves and later in more basal regions of the leaf blade. The KOR3::gusA construct is expressed in the trichome support cells that form a ring at the base of each trichome and in the bundle sheath cells which surround the vascular bundle within the leaf mesophyll tissue. Reverse transcription-polymerase chain reaction of Arabidopsis RNA confirmed the expression of KOR2::gusA and KOR3::gusA. In conclusion, although KOR2 and KOR3 have more restricted expression patterns than the previously characterized KOR gene, they are expressed in cell types at time points where cell wall assembly is likely to occur and, interestingly, differentially expressed in leaf trichomes and their support cells. PMID- 11488475 TI - Molecular characterization and gene expression of lhcb5 gene encoding CP26 in the light-harvesting complex II of Chlamydomonas reinhardtii. AB - We isolated and sequenced a cDNA clone encoding a minor chlorophyll a/b-binding protein, CP26, which is associated with the light-harvesting complex II of Chlamydomonas reinhardtii. Protein sequences of internal peptide fragments from purified CP26 were determined and used to identify a cDNA clone. The 1.1 kb lhcb5 gene codes for a polypeptide of 289 amino acids with a predicted molecular weight of 30,713. The lhcb5 gene product could reconstitute with chlorophylls and xanthophylls to form a green band on a gel. Although the expression of many lhcb genes are strictly regulated by light, the lhcb5 gene was only loosely regulated. We propose that a plant acclimatizes itself to the light environment by quantitatively and qualitatively modulating the light-harvesting complex. Characterization of the primary structure and the implications of its unique expression are discussed. PMID- 11488476 TI - Expression pattern of transcripts encoding water channel-like proteins in Norway spruce (Picea abies). AB - With the aim of identifying cells and tissues with high expression of aquaporins (water channels) or homologous genes in Norway spruce (Picea abies), we report the expression patterns of such transcripts in seedlings, in roots of various ages, and in needles. In situ hybridization experiments with a conserved area of a tonoplast intrinsic protein (TIP) gene from P. abies gave high expression signals in differentiating vascular tissues and in the columella cells of the seedling root cap. High-staining signals were also seen in guard cells and in the bundle sheath cells of needles. Moreover, a slightly increased staining signal was seen in cells forming lateral roots as well as in adventitious roots formed from hypocotyl cuttings. By using PCR-based procedures we also identified a full length aquaporin-like cDNA (mipr) from roots of two-week old seedlings. Sequence homology analysis of the gene suggests that it belongs to the TIP subgroup within the large MIP (major intrinsic protein) family. A phylogenetic analysis of the plant MIP family, including both plasmamembrane (PIP) and tonoplast intrinsic protein (TIP) from Picea, suggests that MIP subgroups evolved already 330 million years ago, as this is the dating of conifer and angiosperm divergence. PMID- 11488477 TI - Tissue-specific signal(s) activate the promoter of a metallocarboxypeptidase inhibitor gene family in potato tuber and berry. AB - The molecular basis of the differential expression of the GM7-type metallocarboxypeptidase inhibitor (MCPI) genes in tuberizing (StMCPI) and non tuberizing Solanum species (SbMCPI) was investigated. It was shown that the StMCPI is encoded by a gene family in Solanum tuberosum (potato), but SbMCPI might be a single-copy gene in the non-tuberizing species Solanum brevidens. The StMCPI promoter shows evolutionary relatedness to the S. brevidens-derived SbMCPI and to the fruit-specific tomato promoter 2A11. Both StMCPI and SbMCPI promoter regions were able to confer tuber- and berry-specific expression for the beta glucuronidase reporter gene in potato suggesting that the difference in MCPI gene expression is in trans regulatory factors between the tuberizing and the non tuberizing Solanum species. The MCPI promoters did not respond to metabolic, environmental or hormonal signals in leaves. Thus, the MCPI genes are regulated in a different way than the other known tuber-specific genes and potentially are suitable for biotechnological application in potato to provide specific transgene expression in tuber and berry. PMID- 11488478 TI - Inducible expression of bacterio-opsin in transgenic tobacco and tomato plants. AB - The development of new strategies to enhance resistance of plants to pathogens is instrumental in preventing agricultural losses. Lesion mimic, the spontaneous formation of lesions resembling hypersensitive response lesions in the absence of a pathogen, is a dramatic phenotype occasionally induced upon expression of certain transgenes in plants. These transgenes simulate the presence of a pathogen and, therefore, activate the plant anti-pathogen defense mechanisms and induce a state of systemic resistance. Lesion mimic genes have been successfully used to enhance the resistance of a number of different plants to pathogen attack. However, constitutive expression of these genes in plants is associated with the spontaneous formation of lesions on leaves and stems, reduced growth, and lower yield. We tested the possibility of using a wound-inducible promoter to control the expression of bacterio-opsin (bO), a transgene that confers a lesion mimic phenotype in tobacco and tomato plants when constitutively expressed. We found that plants with inducible expression of bO did not develop spontaneous lesions. Nevertheless. under controlled laboratory conditions, they were found to be resistant to infection by pathogens. The activation of defense mechanisms by the bO gene was not constitutive, and occurred in response to wounding or pathogen infection. Furthermore, wounding of transgenic tobacco plants resulted in the induction of systemic resistance to pathogen attack within 48 h. Our findings provide a promising initial assessment for the use of wound-inducible promoters as a new strategy to enhance pathogen resistance in transgenic crops by means of lesion mimic genes. PMID- 11488479 TI - The 3' non-coding region of a C4 photosynthesis gene increases transgene expression when combined with heterologous promoters. AB - The Me1 gene of the dicot Flaveria bidentis encodes NADP malic enzyme, which catalyses the decarboxylation reaction of C4 photosynthesis in bundle sheath cells. We have previously shown that the 3' non-coding region (Me1 3') controls quantitative expression of the gene. We wondered whether Me1 3' can increase expression when combined with heterologous promoters. We tested a highly expressed, constitutive promoter, the S4 promoter from subterranean clover stunt virus, and a highly expressed, leaf-specific promoter, the light-harvesting chlorophyll a/b-binding protein gene 3 (Lhcb3) promoter of Arabidopsis thaliana. Promoter-3'-end combinations were tested in transgenic C4 Flaveria plants and C3 tobacco. We found that Me1 3' increased expression of the gusA reporter gene several-fold in leaves of both species in combination with either of the promoters. In both cases Me1 3' does not alter the expression pattern for either promoter. We conclude that Me1 3' can be used as a transcription terminator to increase transgene expression in C3 dicot plants. PMID- 11488480 TI - Two S-adenosylmethionine synthetase-encoding genes differentially expressed during adventitious root development in Pinus contorta. AB - Two S-adenosylmethionine synthetase (SAMS) cDNAs, PcSAMS1 and PcSAMS2, have been identified in Pinus contorta. We found that the two genes are differentially expressed during root development. Thus, PcSAMS1 is preferentially expressed in roots and exhibits a specific expression pattern in the meristem at the onset of adventitious root development, whereas PcSAMS2 is expressed in roots as well as in shoots and is down-regulated during adventitious root formation. The expression of the two SAMS genes is different from the SAMS activity levels during adventitious root formation. We conclude that other SAMS genes that remain to be characterized may contribute to the observed SAMS activity, or that the activities of PcSAMS1 and PcSAMS2 are affected by post-transcriptional regulation. The deduced amino acid sequences of PcSAMS1 and PcSAMS2 are highly divergent, suggesting different functional roles. However, both carry the two perfectly conserved motifs that are common to all plant SAMS. At the protein level, PcSAMS2 shares about 90% identity to other isolated eukaryotic SAMS, while PcSAMS1 shares less than 50% identity with other plant SAMS. In a phylogenetic comparison, PcSAMS1 seems to have diverged significantly from all other SAMS genes. Nevertheless, PcSAMS1 was able to complement a Saccharomyces cerevisiae sam1 sam2 double mutant, indicating that it encodes a functional SAMS enzyme. PMID- 11488481 TI - A Kunitz trypsin inhibitor gene family from trembling aspen (Populus tremuloides Michx.): cloning, functional expression, and induction by wounding and herbivory. AB - Three Kunitz trypsin inhibitor genes were isolated from trembling aspen (Populus tremuloides) by PCR and cDNA screening. Based on sequence similarity, they were grouped into two classes. Southern blots showed complex banding patterns and a high level of restriction fragment polymorphism between different aspen genotypes, suggesting that these trypsin inhibitors are members of a large, rapidly evolving gene family. One of the trypsin inhibitor genes, PtTI2. was over expressed in Escherichia coli and its product shown to inhibit bovine trypsin in vitro. Both classes of PtTI genes are induced by wounding and herbivory, permitting rapid adaptive responses to herbivore pressure. The response appears to be mediated by an octadecanoid-based signaling pathway, as methyl jasmonate treatments induced the trypsin inhibitors. Wound-induced accumulation of trypsin inhibitor protein was also observed by western blot analysis. The pattern of expression, the apparent rapid evolution of TI genes, and the in vitro trypsin inhibitory activity are consistent with a role in herbivore defense. This work establishes the presence of a functional protein-based inducible defense system in trembling aspen. PMID- 11488483 TI - Monochromatic aberrations of the human eye in a large population. AB - From both a fundamental and a clinical point of view, it is necessary to know the distribution of the eye's aberrations in the normal population and to be able to describe them as efficiently as possible. We used a modified Hartmann-Shack wave front sensor to measure the monochromatic wave aberration of both eyes for 109 normal human subjects across a 5.7-mm pupil. We analyzed the distribution of the eye's aberrations in the population and found that most Zernike modes are relatively uncorrelated with each other across the population. A principal components analysis was applied to our wave-aberration measurements with the resulting principal components providing only a slightly more compact description of the population data than Zernike modes. This indicates that Zernike modes are efficient basis functions for describing the eye's wave aberration. Even though there appears to be a random variation in the eye's aberrations from subject to subject, many aberrations in the left eye were found to be significantly correlated with their counterparts in the right eye. PMID- 11488484 TI - What shadows reveal about object structure. AB - In a scene observed from a fixed viewpoint, the set of shadow boundaries in an image changes as a point light source (nearby or at infinity) assumes different locations. We show that for any finite set of point light sources illuminating an object viewed under either orthographic or perspective projection, there is an equivalence class of object shapes having the same set of shadows. Members of this equivalence class differ by a four-parameter family of projective transformations, and the shadows of a transformed object are identical when the same transformation is applied to the light source locations. Under orthographic projection, this family is the generalized bas-relief (GBR) transformation, and we show that the GBR transformation is the only family of transformations of an object's shape for which the complete set of imaged shadows is identical. Finally, we show that given multiple images under differing and unknown light source directions, it is possible to reconstruct both an object's surface and the light source locations up to this family of transformations from the shadows alone. PMID- 11488482 TI - Characterization of two thioredoxins h with predominant localization in the nucleus of aleurone and scutellum cells of germinating wheat seeds. AB - Two full-length cDNA clones, designated TrxhA and TrxhB, encoding different but very similar thioredoxin h polypeptides were isolated from wheat (Triticum aestivum cv. Chinese Spring) aleurone cells. The deduced proteins show a high similarity to each other and to thioredoxin h from other sources, in particular from T. aestivum and T. durum. One of them, TRXhA, was expressed in E. coli as a His-tagged polypeptide and used to raise polyclonal antibodies by immunization of rabbits. These antibodies identified a single band (ca. 13.5 kDa) in western blot analysis of protein extracts from all wheat organs analyzed. TRXhA and TRXhB when expressed in E. coli as intact polypetides showed indistinguishable electrophoretic mobility, which corresponded to the 13.5 kDa polypeptide detected in wheat protein extracts. The amount of thioredoxin h transcripts increased in scutellum and aleurone cells during germination but GA3 did not exert any stimulatory effect on thioredoxin h expression. Although northern blot analysis detected a single band, competitive RT-PCR showed that this band is due to the accumulation of both TrxhA and TrxhB mRNAs. These results suggest that the single band detected in western blots is due to the presence of at least two thioredoxin h polypeptides. Immunolocalization experiments confirmed the high content of thioredoxins h in scutellum and aleurone cells, and showed a low content in the starchy endosperm of germinating grains. Interestingly, though these proteins are evenly distributed in the cytosol, the highest levels of thioredoxins h were detected in the nucleus, both in aleurone and scutellum cells. PMID- 11488485 TI - Analytical model of a three-dimensional integral image recording system that uses circular- and hexagonal-based spherical surface microlenses. AB - A mathematical model for a three-dimensional omnidirectional integral recording camera system that uses either circular- or hexagonal-based spherical surface microlens arrays is derived. The geometry of the image formation and recording process is fully described. Matlab is then used to establish the number of recorded micro-intensity distributions representing a single object point and their dependence on spatial position. The point-spread function for the entire optical process for both close and remote imaging is obtained, and the influence of depth on the point-spread dimensions for each type of microlens and imaging condition is discussed. Comparisons of the two arrangements are made, based on the illustrative numerical results presented. PMID- 11488486 TI - Generalized circular autoregressive models for isotropic and anisotropic Gaussian textures. AB - A new class of random field models, called generalized circular autoregressive (GCAR) models, is introduced. GCAR models have noncausal neighbors that have the same autoregressive parameter values if they are on the same circle or ellipse and that have circular or elliptical correlation structures. This model is better for modeling isotropic or anisotropic natural textures than earlier approaches to modeling of isotropic textures and can represent complex textures with a small number of parameters. Parameter estimation is also considered, and a multistep estimation algorithm is presented. Properties of estimators of GCAR models are also investigated. The efficacy of GCAR models in modeling real textures is demonstrated by synthesizing images resembling real textures by use of parameters estimated from textures selected from the Brodatz texture album. Limitations of GCAR models are also discussed. PMID- 11488487 TI - Degree of nonlinearity and a new solution procedure in scalar two-dimensional inverse scattering problems. AB - Within the framework of inverse scattering problems, the quantifying of the degree of nonlinearity of the problem at hand provides an interesting possibility for evaluating the validity range of the Born series and for quantifying the difficulty of both forward and inverse problems. With reference to the two dimensional scalar problem, new tools are proposed that allow the determination of the degree of nonlinearity in scattering problems when the maximum value, dimensions, and spatial-frequency content of the unknown permittivity are changed at the same time. As such, the proposed tools make it possible to identify useful guidelines for the solution of both forward and inverse problems and suggest an effective solution procedure for the latter. Numerical examples are reported to confirm the usefulness of the tools introduced and of the procedure proposed. PMID- 11488488 TI - Optimum receivers for pattern recognition in the presence of Gaussian noise with unknown statistics. AB - We develop algorithms to detect a known pattern or a reference signal in the presence of additive, disjoint background, and multiplicative white Gaussian noise with unknown statistics. The presence of three different types of noise processes with unknown statistics presents difficulties in estimating the unknown parameters. The standard methods such as expected-maximization-type algorithms are iterative, and in the framework of hypothesis testing they are time consuming, because corresponding to each hypothesis one must estimate a set of parameters. Other standard methods such as setting the gradient of the likelihood function with respect to the unknown parameters will lead to a nonlinear system of equations that do not have a closed-form solution and require iterative methods. We develop an approach to overcome these handicaps and derive algorithms to detect a known object. We present new methods to estimate unknown parameters within the framework of hypothesis testing. The methods that we present are direct and provide closed-form estimates of the unknown parameters. Computer simulations are used to show that for the images tested, the receivers that we have designed perform better than existing receivers. PMID- 11488489 TI - Alternative implementations of the Two-Mu algorithm. AB - I describe implementations of the Two-Mu image-restoration algorithm that model the center portion of the convolution of the point-spread function and the original image (this has been done heretofore), as well as those that model the full range of that convolution. The full convolution methods produce processed images of simple, simulated scenes that are comparable in quality with, and often involve computations that are considerably shorter than, those of the center convolution methods. The full convolution methods incur some loss of information near the edge of the scene. However, that loss may not be significant for large images, especially for those in which the important information is far from the edge of the scene. PMID- 11488490 TI - Natural demodulation of two-dimensional fringe patterns. I. General background of the spiral phase quadrature transform. AB - It is widely believed, in the areas of optics, image analysis, and visual perception, that the Hilbert transform does not extend naturally and isotropically beyond one dimension. In some areas of image analysis, this belief has restricted the application of the analytic signal concept to multiple dimensions. We show that, contrary to this view, there is a natural, isotropic, and elegant extension. We develop a novel two-dimensional transform in terms of two multiplicative operators: a spiral phase spectral (Fourier) operator and an orientational phase spatial operator. Combining the two operators results in a meaningful two-dimensional quadrature (or Hilbert) transform. The new transform is applied to the problem of closed fringe pattern demodulation in two dimensions, resulting in a direct solution. The new transform has connections with the Riesz transform of classical harmonic analysis. We consider these connections, as well as others such as the propagation of optical phase singularities and the reconstruction of geomagnetic fields. PMID- 11488491 TI - Natural demodulation of two-dimensional fringe patterns. II. Stationary phase analysis of the spiral phase quadrature transform. AB - Utilizing the asymptotic method of stationary phase, I derive expressions for the Fourier transform of a two-dimensional fringe pattern. The method assumes that both the amplitude and the phase of the fringe pattern are well-behaved differentiable functions. Applying the limits in two distinct ways, I show, first, that the spiral phase (or vortex) transform approaches the ideal quadrature transform asymptotically and, second, that the approximation errors increase with the relative curvature of the fringes. The results confirm the validity of the recently proposed spiral phase transform method for the direct demodulation of closed fringe patterns. PMID- 11488492 TI - Generality of matched filtering and minimum Euclidean distance projection for optical pattern recognition. AB - Matched filtering followed by a minimum Euclidean distance projection onto realizable filter values was previously shown to optimize the signal-to-noise ratio for single training images in optical correlation pattern recognition. The algorithm is now shown to solve the combination of (1) standard statistical pattern-recognition metrics with multiple training images, (2) additive input noise of known power spectral density and also additive detection noise that is irreducible by the filter, (3) the building of the filter on arbitrary subsets of the complex unit disk, and (4) the use of observable correlator outputs only. The criteria include the Fisher ratio, the Bayes error and Bayes cost, the Chernoff and Bhattacharyya bounds, the population entropy and expected information, versions of signal-to-noise ratio that use other than second power in their norm, and the area under the receiver operating characteristic curve. Different criteria are optimized by different complex scalar weights. PMID- 11488493 TI - Propagation of flat-topped multi-Gaussian laser beams. AB - The multi-Gaussian beam shape is proposed as a model for aperture functions and laser beam profiles that have a nearly flat top but whose sides decrease continuously. Beams and apertures of this type represent a simple, elegant, and intuitive alternative to super-Gaussian beams, which are important in a number of applications such as laser resonator design. Analytical formulas are developed for the propagation of these beams through free space and optical systems representable by ABCD matrices. PMID- 11488494 TI - Helical localized wave solutions of the scalar wave equation. AB - A right-handed helical nonorthogonal coordinate system is used to determine helical localized wave solutions of the homogeneous scalar wave equation. Introducing the characteristic variables in the helical system, i.e., u = zeta - ct and v = zeta + ct, where zeta is the coordinate along the helical axis, we can use the bidirectional traveling plane wave representation and obtain sets of elementary bidirectional helical solutions to the wave equation. Not only are these sets bidirectional, i.e., based on a product of plane waves, but they may also be broken up into right-handed and left-handed solutions. The elementary helical solutions may in turn be used to create general superpositions, both Fourier and bidirectional, from which new solutions to the wave equation may be synthesized. These new solutions, based on the helical bidirectional superposition, are members of the class of localized waves. Examples of these new solutions are a helical fundamental Gaussian focus wave mode, a helical Bessel Gauss pulse, and a helical acoustic directed energy pulse train. Some of these solutions have the interesting feature that their shape and localization properties depend not only on the wave number governing propagation along the longitudinal axis but also on the normalized helical pitch. PMID- 11488495 TI - Guided-mode resonant subwavelength gratings: effects of finite beams and finite gratings. AB - The effects of finite beams and finite gratings on the performance of guided-mode resonant subwavelength gratings are characterized by using the rigorous boundary element method. The gratings are strongly modulated, have a finite number of periods, and are illuminated by normally incident Gaussian beams. Quantitative results are presented for silicon-on-sapphire resonant gratings and gallium arsenide-aluminum arsenide resonant gratings. PMID- 11488496 TI - On the extinction of radiation by a homogeneous but spatially correlated random medium. AB - Exponential extinction of incoherent radiation intensity in a random medium (sometimes referred to as the Beer-Lambert law) arises early in the development of several branches of science and underlies much of radiative transfer theory and propagation in turbid media with applications in astronomy, atmospheric science, and oceanography. We adopt a stochastic approach to exponential extinction and connect it to the underlying Poisson statistics of extinction events. We then show that when a dilute random medium is statistically homogeneous but spatially correlated, the attenuation of incoherent radiation with depth is often slower than exponential. This occurs because spatial correlations among obstacles of the medium spread out the probability distribution of photon extinction events. Therefore the probability of transmission (no extinction) is increased. PMID- 11488497 TI - Optical radiation flux illuminating a circular disk from an off-axis point source separated by a homogeneous plane-parallel plate. AB - In a number of techniques that measure weak fluxes of optical radiation, it is frequently necessary to keep a detector in a medium different from that of the radiating source by separating it from the source with a planar transparent window. However, sources such as systems of light-emitting diodes, large-fiber illuminators, and microscopic living objects that emit biological luminescence may sometimes be regarded as multiple-point sources. To estimate the fluxes of optical radiation illuminating a surface from a nonuniformly distributed multiple point source, a method for calculating fluxes from a single off-axis point source is needed. A formula is derived to estimate a flux of temporally incoherent optical radiation incident on a circular disk from a single off-axis point source separated by a plane-parallel plate (PPP). This formula is expressed by a series of single integrals of some superposed elementary functions. These functions depend on the variables that characterize the point-source-plane-parallel-plate circular-disk geometry and on the optical properties of the media that separate the source from the PPP and the PPP from the disk. The solution was obtained for isotropic media. For illustrative purposes some examples of the use of the formula are presented. The selected results are illustrated by three-dimensional surface plots and compared with the values of the fluxes calculated for radiation incident on the disk from a point source not separated by a PPP. PMID- 11488498 TI - Radiation forces in the discrete-dipole approximation. AB - The theory of the discrete-dipole approximation (DDA) for light scattering is extended to allow for the calculation of radiation forces on each dipole in the DDA model. Starting with the theory of Draine and Weingartner [Astrophys. J. 470, 551 (1996)] we derive an expression for the radiation force on each dipole. These expressions are reformulated into discrete convolutions, allowing for an efficient, O(N logN) evaluation of the forces. The total radiation pressure on the particle is obtained by summation of the individual forces. The theory is tested on spherical particles. The resulting accumulated radiation forces are compared with Mie theory. The accuracy is within the order of a few percent, i.e., comparable with that obtained for extinction cross sections calculated with the DDA. PMID- 11488499 TI - Model acquisition and invariant tracking of unknown materials in hyperspectral images. AB - We consider the problem of acquiring models for unknown materials in airborne 0.4 2.5 microm hyperspectral imagery and using these models to identify the unknown materials in image data obtained under significantly different conditions. The material models are generated with use of an airborne sensor spectrum measured under unknown conditions and a physical model for spectral variability. For computational efficiency, the material models are represented by using low dimensional spectral subspaces. We demonstrate the effectiveness of the material models by using a set of material tracking experiments in HYDICE images acquired in forest and desert environments over widely varying conditions. We show that techniques based on the new representation significantly outperform methods based on direct spectral matching. PMID- 11488500 TI - Detection of inhomogeneities with ultrasound tagging of light. AB - Ultrasound modulated light for optical tomography is very useful, since it can provide three-dimensional data with minimal mathematical processing. Although several experimental studies have shown the potential of this method, the link between the ultrasound location and the modulated signal intensity at the detector is not yet fully understood. We derive an analytical formula relating the position of the ultrasound transducer and the optical signal at the detector. We also derive an expression for the signal-to-shot-noise ratio as a function of the transducer position. We show that in certain conditions this ratio is only slowly decreasing as a function of the light penetration depth, which makes this technique attractive for optical tomography. PMID- 11488501 TI - Angle eikonals for a perfect zoom system. AB - I show that it is possible to choose the eikonals for the lens groups of a zoom lens such that extended objects at infinity are imaged perfectly, without any aberrations, at all zoom settings. PMID- 11488502 TI - Phase-map measurements by interferometry with sinusoidal phase modulation and four integrating buckets. AB - Phase-shifting interferometry based on the integrating-bucket technique with sinusoidal phase modulation is studied theoretically and demonstrated experimentally to obtain phase maps from double-beam interferometers. The method uses four frames obtained by integration of the time-varying intensity in an interference pattern during the four quarters of the modulation period. An optimum sinusoidal phase modulation is found to minimize the effect of the additive noise. The absolute accuracy of the phase measurements is discussed. Possible applications of the method are demonstrated with two interference microscopes with which the phase modulation is achieved by sinusoidal oscillation of a mirror attached to a piezoelectric transducer and by sinusoidal birefringence modulation with a photoelastic modulator. In both experimental arrangements, phase images can be produced in real time at a rate of several hertz. Noise measurements are reported and compared with theory. PMID- 11488503 TI - Alignment and calibration of the MgF2 biplate compensator for applications in rotating-compensator multichannel ellipsometry. AB - Biplate compensators made from MgF2 are being used increasingly in rotating element single-channel and multichannel ellipsometers. For the measurement of accurate ellipsometric spectra, the compensator must be carefully (i) aligned internally to ensure that the fast axes of the two plates are perpendicular and (ii) calibrated to determine the phase retardance delta versus photon energy E. We present alignment and calibration procedures for multichannel ellipsometer configurations with special attention directed to the precision, accuracy, and reproducibility in the determination of delta (E). Run-to-run variations in external compensator alignment, i.e., alignment with respect to the incident beam, can lead to irreproducibilities in delta of approximately 0.2 degrees . Errors in the ellipsometric measurement of a sample can be minimized by calibrating with an external compensator alignment that matches as closely as possible that used in the measurement. PMID- 11488504 TI - Axicon-based Bessel resonator: analytical description and experiment. AB - We present a new scheme for an optical resonator for production of Bessel and Bessel-Gauss light beams. The resonator with Bessel modes is composed of two plane mirrors with an axicon placed close to one of them. If this mirror is concave, the modes are Bessel-Gauss light beams. Analytical expressions relating parameters of the resonator and characteristics of its modes are obtained and analyzed. The results are verified with the Fox-Li algorithm. The resonator scheme was implemented in an experiment to confirm the possibility of the generation of zero-order Bessel beams. It was found that multipass modes can also oscillate in the resonator if its apertures are large enough. PMID- 11488505 TI - Integral equations applied to wave propagation in two dimensions: modeling the tip of a near-field scanning optical microscope. AB - We present a Green's-function/Green's-theorem integral equation approach to numerically modeling two-dimensional, s-polarized, wave propagation problems effectively for a variety of geometries. The model accurately calculates both near fields and far fields because of the minimal assumptions made on the behavior of the scattered radiation. The method was applied to modeling light emission from a near-field scanning optical microscope fiber tip. Several convergence and energy tests were used to give confidence in the results. The behavior of intensity and power near the tip was investigated. The effects of changing the dielectric constant of a sample material located below the tip were also examined. PMID- 11488506 TI - Three-dimensional coherent transfer function for reflection confocal microscopy in the presence of refractive-index mismatch. AB - The three-dimensional (3-D) coherent transfer function for reflection confocal microscopy of high-numerical-aperture objectives is derived and calculated in the presence of refractive-index mismatch when a laser beam is focused into a medium of refractive index different from its immersion medium. This aberrated coherent transfer function is then used to estimate the readout efficiency of 3-D data bits recorded in a thick medium. It is shown that the readout efficiency of confocal microscopy for 3-D bit data storage is decreased with the focal depth of an objective in a recording medium. However, a high readout efficiency can be maintained if the tube length of a reading objective is linearly altered to compensate for the spherical aberration caused by the refractive-index mismatch. PMID- 11488507 TI - Beam propagation through uniaxial anisotropic media: global changes in the spatial profile. AB - The propagation of electromagnetic beams through uniaxial anisotropic media is investigated. The Maxwell equations are solved in the paraxial limit in terms of the plane-wave spectrum associated with each Cartesian field component. Attention is focused on the global changes in the spatial structure of the beam, which are described by means of the second-order intensity moment formalism. In particular, the propagation law for the intensity moments through this kind of media is obtained. As a consequence it is inferred that it is possible to improve the beam quality parameter by using these media. PMID- 11488508 TI - Comparison of highly efficient absorbing boundary conditions for the beam propagation method. AB - Numerical experiments using the paraxial finite-difference beam propagation method have been performed with the following boundary conditions: perfectly matched layer, Higdon absorbing boundary conditions, complementary operators method, and extended complementary operators method. We have shown that Higdon operators must be modified for the paraxial wave equation to take into account the spectrum of incident rays on the boundaries of the computational domain. Reflection coefficients, accuracy, numerical dissipation/ gain, memory requirements, and time computation are compared and discussed for these absorbing techniques. PMID- 11488509 TI - Changes in the metal content of surficial sediments of Boston Harbor since the cessation of sludge discharge. AB - Temporal trends of metals in surficial sediments (1991-1998) at two sites in Boston Harbor were analyzed to evaluate the effect of stopping sludge dumping in December 1991. Metal contents of sediments from the old sludge disposal site were higher than those of a station in the central Harbor. Since 1991, carbon, copper, and lead contents have significantly decreased in sediments from the disposal site. Chromium and Zn have shown smaller decreases while Fe, and Mn, have remained relatively constant. Metal content in the central Harbor station, located in an area of sediment reworking, has been quite variable, but, with the exception of Zn which has shown a large decrease relative to iron, the changes seemed to be well correlated with changes in the organic carbon content at this site due to resuspension. Ratios of metals in the sediments are fairly similar at both sites and similar to those of sewage-derived particles, with the exception of Cr, which appears to be enriched in the sediments. PMID- 11488510 TI - Availability of cadmium and zinc from sewage sludge to the flounder, Platichthys flesus, via a marine food chain. AB - This paper examines the potential availability to a demersal fish of cadmium and zinc associated with digested sewage sludge, via a food chain as well as directly from the sludge, and the tissue distribution and possible excretion of any accumulated cadmium and zinc. Radioactive tracer techniques were used in order to follow the food chain transfer of the metals. Flounder (Platichthys flesus) accumulated cadmium both directly from sludge (delivered in newly ingested unassimilated gutfuls in the amphipods) and in assimilated form from the tissues of the amphipod crustacean Corophium volutator that had been feeding on a sludge sediment mixture. Cadmium from both sources could be excreted by the fish. The concentration of cadmium within the whole fish increased as the input of cadmium in the diet increased. Zinc, similarly, is available to the flounder both directly from sludge and from zinc accumulated in C. volutator. Only a limited increase in net accumulation of zinc by the flounder was observed upon increased inputs of zinc through the diet, perhaps indicating some regulation of body zinc concentration by the flounder. PMID- 11488511 TI - Barnacles as biomonitors of trace metal availabilities in Hong Kong coastal waters: changes in space and time. AB - The use of selected organisms as biomonitors of trace metal bioavailabilities allows comparisons to be made over space and time. The concentrations of 11 trace metals (arsenic, cadmium, chromium, cobalt, copper, iron, lead, manganese, nickel, silver, zinc) were measured in the bodies of two barnacle species, Balanus amphitrite and Tetraclita squamosa, from up to 18 littoral sites from Hong Kong coastal waters in April 1998. These data provide evidence on the geographical variation in metal bioavailabilities at this time, and are compared selectively against historical data sets for 1986 and 1989. Geographical variation in bioavailabilities is clear for several metals, with hotspots for arsenic, copper, nickel and silver at Chai Wan Kok, and for lead in Junk Bay. Victoria Harbour sites head the rankings for silver and arsenic, and Tolo Harbour sites exhibit relatively elevated cobalt, manganese and zinc. Many bioavailabilities of trace metals to barnacles are lower in Hong Kong coastal waters in 1998 than in 1986. The two barnacle species are widespread and the extensive data set presented is a benchmark which can be compared to the results of similar biomonitoring programmes elsewhere in the Indo-Pacific and beyond. PMID- 11488512 TI - Distribution of cytochrome P4501A (CYP1A) in the tissues of Baltic ringed and grey seals. AB - Information about the expression of CYP1A in wildlife species is essential for understanding the impact of organochlorine exposure on the health status of an exposed population. Therefore, we aimed at characterising a putative CYP1A enzyme expression in both hepatic and extrahepatic tissues of ringed and grey seals from the Baltic Sea and from less polluted waters. The cellular localisation of CYP1A was identified using a monoclonal antibody against scup P4501A1 (MAb 1-12-3). Immunohistochemical staining showed the highest level of CYP1A expression in liver hepatocytes, and the second highest level in the endothelial cells of capillaries and larger blood vessels in the liver and other organs. The most frequent and strongest staining was found in Baltic ringed seals. Although CYP1A positive staining was observed in only a few tissues in the other seal populations, it was more intense in Baltic grey seals than in Canadian grey seals. The CYP1A enzyme activity, expressed as ethoxyresorufin O-deethylation (EROD), followed a similar tissue distribution and geographical pattern as the immunohistochemistry with clearly elevated EROD activities in most tissues of both Baltic seal populations. Immunochemical characterisation by immunoblotting confirmed the presence and elevation pattern of a putative CYP1A protein in ringed and grey seals, supporting our findings using other methods. The evenly distributed elevation of CYP1A expression among most of the tissues examined indicates that Baltic seals are exposed to CYP1A inducing agents affecting the whole body. This may result in an increased or decreased toxic potential of foreign substances, which may ultimately determine the biological effects of the contaminants. PMID- 11488513 TI - Can you do a community assessment without talking to the community? AB - Conventionally, it has been accepted that an accurate understanding of community views on health topics can only come directly from community members. Yet, even as the need for this data has been increased by managed care, the methods of gathering it from community members remain costly and impractical for most busy practices or communities. This case study was conducted to determine whether well informed health professionals can accurately and less expensively convey community views on health topics. In a low-income urban community, using a standard set of open-ended questions, focus groups and telephone and mail surveys were used to gather qualitative data from community residents and patients. Extended interviews and key informant surveys were used to gather similar data from health professionals. Data obtained from both sets of respondents were compared for content and logistics of collection. Overall, responses from the two groups had similar content but differing emphases. Community residents tended to emphasize socioeconomic determinants of health and everyday life concerns, while health care workers tended to emphasize the process of care. However, these perspectives were shared across the groups, as was an emphasis on the need for education with regard to community health concerns. The most striking difference between the groups was the community resources each group identified. As expected, costs for the health care worker approaches were less expensive. Findings from this case study suggest limited situations in which health care workers can adequately convey community views. In other situations, the different perspectives of each group can be complementary. Regardless of their role in qualitative data gathering, there remain important roles for community members in planning and decision-making regarding their health care. PMID- 11488514 TI - Mammography utilization pattern throughout the state of Arkansas: a challenge for the future. AB - The Mammography Data Collection Project (MDCP) was aimed at determining the 1997 mammography rate and screening practice patterns throughout the state of Arkansas. The MDCP conducted a comprehensive survey of mammography centers to collect individual patient demographic data, and mammography data. Data elements pertinent to patients' age, race, insurance, type of mammogram, date of mammogram, and the zip code were obtained from participating centers. Analysis was performed to determine the mammography rate by age group and areas of residence. Of the 112 eligible mammography centers contacted, 92 centers (82%) participated. A total of 157,976 mammography data sets were obtained. The total number of women imaged was 148,586 with 90% of them over the age of 40 years. The mammography rate was 24.1% for women 50 years of age and older and 19.4% for women 40-49 years of age. Most of the women imaged had private insurance (52%). This study revealed that only a small fraction of women ages 40 years and older obtained annual mammograms in 1997, thus highlighting the need for intensifying efforts to increase the utilization of this lifesaving test. PMID- 11488515 TI - Community participation in road safety: barriers and enablers. AB - The objectives of the paper were to identify barriers to community participation in road safety activities and to recommend strategies to reduce those barriers. Information was obtained from: a review of relevant literature, from the authors' extensive community-based experiences, from undertaking community-based research, and from interviews with key stakeholders in road safety in Western Australia. Ten significant barriers to community participation were identified. These are classified into two groups: personnel and planning issues. The former includes: reasons why people are often reluctant to become involved in projects in their communities, a lack of leadership, and a lack of skills. The latter includes: inappropriate program foci, inappropriate program evaluation, lack of resources, and a lack of sustainability. It was concluded that the barriers to community involvement in road safety initiatives are similar in many parts of Australia and overseas and apply to a wide range of health promotion interventions besides road safety. Awareness of the barriers and knowledge of their solutions can ensure that appropriate steps are taken to maximise the likelihood of community participation. PMID- 11488516 TI - Public perceptions of child abuse and neglect in a midwestern urban community. AB - This study found that the majority of urban adults did not believe that any of the characteristics of children posed to them predisposed a child to abuse. In contrast, 40% of respondents claimed child abuse could not occur in a family like the one in which they grew up. The majority perceived only one characteristic, alcohol abuse, as a characteristic of child abusers. Few respondents (10%) believed that child abusers could not be helped, and 25% were not sure. Adults' perceptions of child abuse and abusers varied by sex, age, socioeconomic status, and whether the respondents had children. The data indicate that there are significant deficits in the respondents' knowledge of child abuse and neglect. PMID- 11488517 TI - Caregiving, alcohol use, and mental health symptoms among HMO members. AB - Using data from a survey of 5841 HMO members, we examined alcohol consumption, depression and anxiety symptoms, and health services use among 689 informal caregivers. Characteristics of caregivers included whom they cared for, types of care provided, number of people cared for, and whether care was provided in the caregiver's home. Outcome measures in hierarchical linear and logistic stepwise regression models included indicators of alcohol drinking pattern, symptoms of anxiety and depression, role limits due to emotional problems and self-reported doctor's visits. Adjusting for age and gender differences, caregivers reported more bodily pain, worse role functioning related to emotional problems, were more likely to screen as depressed, and were more likely to report symptoms of anxiety. Older caregivers, and those with greater vitality, had better mental health outcomes; caregivers with higher levels of education reported better psychological well-being and less interference with role functioning due to emotional problems. Caregivers did not make more doctor's visits than non caregivers. No caregiving characteristics were consistently associated with both mental health and alcohol consumption outcomes. The relationship of the care recipient to the caregiver predicted mental health outcomes better than either the type of care, number cared for, or location of care. Both the type of care and relationship to the care provider were associated with alcohol consumption, and several gender interactions were identified. Our results confirm previous findings of greater mental health problems among caregivers, yet suggest that caregivers may not be seeking services commensurate to their needs. PMID- 11488518 TI - Expression and integrity of DNA topoisomerase II isoforms does not explain generic drug resistance in malignant mesothelioma. AB - PURPOSE: Malignant mesothelioma is a tumour that is highly resistant to a number of different chemotherapy agents, yet the mechanisms by which resistance occurs are poorly understood. The pattern of resistance is consistent with disruption of topoisomerase function or expression. Coupled with this, we have previously noted a common serological reaction to the beta isoform of topoisomerase II, suggesting that it may be aberrantly expressed in patients with mesothelioma. METHODS: We assessed the expression of topoisomerase II isoforms in sections of primary tumour. We tested a panel of five mesothelioma cell lines for sensitivity to the known topoisomerase-targeting drugs, doxorubicin and etoposide. We sequenced expressed segments of the topoisomerase genes from these cell lines that have previously been associated with drug resistance. We then investigated other potential resistance mechanisms. RESULTS: We found that the beta isoform of topoisomerase II was more frequently expressed in primary tumours. Only one of the five cell lines was highly resistant to etoposide and this cell line was found to have a point mutation in the gene for topoisomerase IIalpha. Protein levels of topoisomerase IIalpha and beta did not correlate with sensitivity to either doxorubicin nor to etoposide. Semiquantitative analysis suggested that there was marked variation in the levels of mRNA expression of MRP, gamma-GCS and MDR1. None of these findings could be associated with resistance to chemotherapy. CONCLUSION: We conclude that mutations in topoisomerase IIalpha can be associated with extreme resistance of mesothelioma to etoposide. The generic drug resistance of this tumour requires further investigation. PMID- 11488519 TI - Pharmacological study of paclitaxel duration of infusion combined with GFR-based carboplatin in the treatment of ovarian cancer. AB - PURPOSE: To determine the effect on systemic pharmacology and clinical toxicity of dose and mode of administration of paclitaxel combined with carboplatin in the treatment of ovarian cancer. PATIENTS AND METHODS: A total of 18 patients were treated with a dose of carboplatin determined by GFR, to attain a target AUC of 6 or 7 mg/ml x min. The paclitaxel dose was 175 or 200 mg/m2 administered over approximately 1 or 3 h. The duration of infusion was randomized, crossing over to the alternative treatment for the second course. Blood samples were analysed for carboplatin, paclitaxel and for the excipients of the paclitaxel formulation, ethanol and Cremophor. RESULTS: Overall the three-weekly schedule of administration of the combination of carboplatin and paclitaxel was well tolerated. There were no clinical differences in the toxicities observed between courses where a 1-h infusion was used compared with those with a 3-h infusion. The target AUC of carboplatin was achieved (mean +/- SD 114 +/- 20% of target). Analysis of paclitaxel pharmacokinetics did not show a difference in the AUC or time above a pharmacological threshold for the two infusion durations. The peak concentration of paclitaxel obtained at the end of the infusion (9.1 vs 4.5 microg/ml), and the plasma ethanol concentration (40.0 vs 20.5 mg/dl) were higher following the shorter duration infusion. Peak concentrations of Cremophor were not different. CONCLUSION: The combination of paclitaxel at a dose of 175 mg/m2 and carboplatin at a target AUC of 6-7 mg/ml min can safely be administered every 3 weeks. Also, a 1-h infusion of paclitaxel has no acute clinical disadvantage over a 3-h infusion and these durations of administration are pharmacologically equivalent. PMID- 11488520 TI - Activity of high-dose toremifene plus cisplatin in platinum-treated non-small cell lung cancer: a phase II California Cancer Consortium Trial. AB - PURPOSE: Although cisplatin is an important agent in non-small-cell lung cancer (NSCLC), de novo resistance is common and acquired resistance emerges rapidly during therapy. Proposed mediators of platinum resistance include the protein kinase C (PKC) signal transduction pathway and associated c-FOS overexpression. While estrogen administration has been reported to upregulate PKC and c-FOS expression, the triphenylethylenes tamoxifen and toremifene potentiate platinum cytotoxicity by inhibition of PKC. Downregulation of c-FOS expression has been reported to result from PKC inhibition. In view of these findings, we hypothesized that toremifene would reverse platinum resistance and that this interaction would be influenced by tumor estrogen receptor (ER) status. MATERIALS AND METHODS: A phase II trial of high-dose toremifene (600 mg orally daily on days 1-7) plus cisplatin (50 mg/m2 intravenously on days 4 and 11) every 28 days in NSCLC patients was conducted. A group of 30 patients with metastatic NSCLC who had been previously treated with platinum-based therapy were enrolled. RESULTS: All of the 30 patients were assessable for toxicity and 28 for tumor response. Therapy was well tolerated with minimal hematologic and non-hematologic toxicity. Common toxicity criteria grade 3 hematologic toxicity was seen in only three patients. Five patients achieved a partial response for an overall response rate of 18% (95% CI 6-37). Median overall survival was 8.1 months (95% CI 5.4-17). To assess PKC, ER, and c-Fos expression by immunohistochemistry, 12 informative pretreatment patient tumor specimens were obtained. Four patient tumor specimens were positive for one or both PKC isoforms (alpha and epsilon) while c-Fos was overexpressed in three. None of the responding patient tumors exhibited c-FOS or PKC-epsilon overexpression. ER expression was found to be infrequent (8%), contrasting with previous reports in this tumor type. CONCLUSION: While this phase II study indicates that high-dose toremifene plus cisplatin is feasible, active, and well tolerated in NSCLC patients previously treated with platinum compounds, the mechanism of action remains unclear. Further study of this regimen is warranted. PMID- 11488521 TI - Structure-activity relationship studies of discodermolide and its semisynthetic acetylated analogs on microtubule function and cytotoxicity. AB - PURPOSE: Discodermolide, a natural product from the marine sponge Discodermia dissoluta, has been previously described as an antimitotic agent with microtubule hyperstabilizing properties similar to those of paclitaxel (Taxol). The clinical success of paclitaxel has led to a growing interest in novel antimitotic compounds and the elucidation of their structure-activity characteristics. Analogs of discodermolide were prepared by acetylation of the hydroxyl groups at carbons 3, 7, 11 and/or 17 and tested for biological activity in human tumor cells to determine the structural requirements for tubulin interaction and cytotoxic effects. METHODS: A549 human lung adenocarcinoma cells were incubated with discodermolide, or its acetylated analogs, and examined for their effects on microtubule architecture, cytotoxicity. and perturbations of the cell cycle. To confirm their direct interaction with tubulin. analogs were assayed for their ability to induce the polymerization of purified bovine brain tubulin. RESULTS: Acetylation of discodermolide at the C-7 hydroxyl group potentiated the cytotoxicity of the molecule to A549 cells, whereas acetylation at the C-3 hydroxyl group had little effect on the cytotoxicity of the parent or C-7 acetylated compounds. The acetylation of the hydroxyl groups at the C-11 and C-17 positions severely abrogated the cytotoxicity of the molecule. Cell cycle analysis by flow cytometry revealed that the more cytotoxic analogs caused the accumulation of cells in the G2/M phase, a mechanism previously reported for discodermolide. All discodermolide analogs with IC50 values below 1000 nM exhibited microtubule effects to varying degrees in cultured A549 cells, yet only the most cytotoxic promoted the polymerization of purified tubulin. CONCLUSIONS: Although the parent compound was more effective at polymerizing purified tubulin, acetylation of the C-3 or C-3 and C-7 hydroxyl groups improved its cytotoxicity in whole cells suggesting that acetylation either enhances accumulation of the molecules within cells or imparts a secondary cytotoxic quality not present in the discodermolide molecule. The study reported here is the first to provide information on the structure-activity relationships of discodermolide using human tumor cells and analogs produced by semisynthetic modification of natural discodermolide. PMID- 11488522 TI - Fendiline, an anti-anginal drug, increases intracellular Ca2+ in PC3 human prostate cancer cells. AB - BACKGROUND: The effects of the anti-anginal drug fendiline on intracellular Ca2+ concentrations ([Ca2+]i) in human PC3 prostate cancer cells were examined. METHODS: [Ca2+]i was measured using the fluorescent dye fura-2. RESULTS: Fendiline (0.5-100 microM) increased [Ca2+]i in a concentration-dependent manner. Ca2+ removal partly inhibited the Ca2+ signals. In Ca2+-free medium, pretreatment with 100 microM fendiline inhibited most of the [Ca2+]i increase induced by 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor), and pretreatment with thapsigargin abolished the fendiline-induced [Ca2+]i increases. Adding 3 mM Ca2+ increased [Ca2+]i in cells pretreated with 0.5-200 microM fendiline in Ca2+-free medium. Pretreatment with 1 microM U73122 to block the formation of inositol-1.4.5-trisphosphate (IP3) did not alter fendiline-induced internal Ca2+ release. CONCLUSIONS: The anti-anginal drug fendiline induced internal Ca2+ release and external Ca2+ entry. Because prolonged increases in [Ca2+]i may lead to cell injury and death, the long-term effect of fendiline on the function of prostate cancer cells should be investigated. PMID- 11488523 TI - Induction of cytochrome P450 3A4 by docetaxel in peripheral mononuclear cells and its expression in lung cancer. AB - Several recent studies have demonstrated that the cytochrome p450 (CYP) family plays an important role in the metabolism of taxanes. However, the role of CYP gene expression in tumors and peripheral mononuclear cells (PMN) is unknown. We therefore investigated the levels of CYP3A4 and CYP2C gene expression using reverse transcription polymerase chain reaction (RT-PCR) in PMN from 16 previously untreated lung cancer patients to determine whether the expression of the two genes is induced by docetaxel (TXT). Neither the CYP3A4 nor the CYP2C gene was induced after administration of carboplatin (CBDCA) alone. Expression of the CYP3A4 gene was induced by the administration of TXT alone or TXT and CBDCA, but expression of the CYP2C gene was unaffected. We also measured the expression of both genes using RT-PCR in 20 autopsy samples (ten non-small-cell lung cancers and their corresponding normal lung tissues) obtained from patients who had not received any chemotherapy during life. The level of CYP2C gene expression in samples of lung cancer was significantly higher than in normal lung tissue, but the level of CYP3A4 gene expression was not. These results suggest that the CYP3A4 gene is induced by TXT, and that it plays an important role in intracellular TXT metabolism. PMID- 11488524 TI - Acyl derivatives of demethylpenclomedine, an antitumor-active, non-neurotoxic metabolites of penclomedine. AB - PURPOSE: The purpose of this investigation was to compare the antitumor activities of a series of acyl derivatives of 4-demethylpenclomedine (DM-PEN), the major plasma metabolite of penclomedine (PEN) observed to be an active antitumor agent in vivo and non-neurotoxic in a rat model with that of DM-PEN. METHODS: Acyl derivatives were prepared from DM-PEN and evaluated in vivo against human MX-1 breast tumor xenografts implanted subcutaneously (s.c.) or intracerebrally (i.c.). Several derivatives were also evaluated against other human tumor xenografts and murine P388 leukemia cell lines. RESULTS: Several of the acyl derivatives were found to be superior to DM-PEN against MX-1, human ZR 75-1 breast tumor, human U251 CNS tumor and the P388 leukemia parent cell line and lines resistant to cyclophosphamide and carmustine. 4-Demethyl-4 methoxyacetylpenclomedine showed inferior activity to current clinical brain tumor drugs against a glioma cell line, superior activity to temozolomide and procarbazine against the derived mismatch repair-deficient cell line, and superior activity to cyclophosphamide and carmustine but inferior activity to temozolomide against two ependymoma cell lines, all of which were implanted s.c. CONCLUSION: Proposed mechanisms of activation and action of DM-PEN and the acyl derivatives support the potential clinical superiority of the acyl derivatives. PMID- 11488525 TI - Population pharmacokinetics of ifosfamide and its 2- and 3-dechloroethylated and 4-hydroxylated metabolites in resistant small-cell lung cancer patients. AB - The aim of this study was to develop a population pharmacokinetic model that could describe the pharmacokinetics of ifosfamide. 2- and 3 dechloroethylifosfamide and 4-hydroxyifosfamide, and calculate their plasma exposure and urinary excretion. A group of 14 patients with small-cell lung cancer received a 1-h intravenous infusion of 2.0 or 3.0 g/m2 ifosfamide over 1 or 2 days in combination with 175 mg/m2 paclitaxel and carboplatin at AUC 6. The concentration-time profiles of ifosfamide were described by an ifosfamide concentration-dependent development of autoinduction of ifosfamide clearance. Metabolite compartments were linked to the ifosfamide compartment enabling description of the concentration-time profiles of 2- and 3 dechloroethylifosfamide and 4-hydroxyifosfamide. The Bayesian estimates of the pharmacokinetic parameters were used to calculate the systemic exposure to ifosfamide and its metabolites for the four ifosfamide schedules. Fractionation of the dose over 2 days resulted increased metabolite formation, especially of 2 dechloroethylifosfamide, probably due to increased autoinduction. Renal recovery was only minor with 6.6% of the administered dose excreted unchanged and 9.8% as dechloroethylated metabolites. In conclusion, ifosfamide pharmacokinetics were described with an ifosfamide concentration-dependent development of autoinduction and allowed estimation of the population pharmacokinetics of the metabolites of ifosfamide. Fractionation of the dose resulted in increased exposure to 2 dechloroethylifosfamide, probably due to increased autoinduction. PMID- 11488526 TI - Markedly diminished drug resistance-inducing properties of vinflunine (20',20' difluoro-3',4'-dihydrovinorelbine) relative to vinorelbine, identified in murine and human tumour cells in vivo and in vitro. AB - PURPOSE: Vinflunine (VFL) is a novel Vinca alkaloid with markedly superior experimental in vivo antitumour activity to its parent molecule, vinorelbine (Navelbine, NVB), against a panel of murine and human tumours. The aim of this study was to establish whether there are differences in the rate and extent of development of resistance, both in vivo and in vitro, to these two newer Vinca alkaloids under identical selection conditions. METHODS: Using P388 leukaemia cells in vivo, it was evident that VFL induced drug resistance far less readily than NVB, as shown by the number of passages required to select for total resistance. Under in vitro conditions, using A549 human lung carcinoma cells, it was also clearly shown by drug sensitivity determinations that VFL was a less potent inducer of drug resistance than NVB. Resistance resulting from either in vivo or in vitro selection was associated with a classic multidrug resistance profile. Further characterization of the drug-resistance phenotype of the most highly resistant A549 sublines showed that the level of total beta-tubulin expression appeared to be modified exclusively in the NVB-resistant cells. CONCLUSION: The clear demonstration that resistance to VFL developed far less readily than resistance to NVB both in vivo and in vitro may have potential clinical implications. PMID- 11488527 TI - Antineoplastic action of 5-aza-2'-deoxycytidine and histone deacetylase inhibitor and their effect on the expression of retinoic acid receptor beta and estrogen receptor alpha genes in breast carcinoma cells. AB - PURPOSE: During tumorigenesis several cancer-related genes can be silenced by aberrant methylation. In many cases these silenced genes can be reactivated by exposure to the DNA methylation inhibitor, 5-aza-2'-deoxycytidine (5-AZA-CdR). Histone acetylation also plays a role in the control of expression of some genes. The aim of this study was to determine the antineoplastic activities of 5-AZA-CdR and trichostatin A (TSA), either administered alone or in combination. in MDA-MB 231 breast carcinoma cells. The effects of these drugs (alone and in combination) on the expression of the tumor suppressor gene, retinoic acid receptor (RAR beta) and of the estrogen receptor alpha gene (ER alpha), whose expression is lost in the cell line used in the study, were also investigated. METHODS: MDA-MB-231 cells were treated with 5-AZA-CdR and TSA and the antitumor activity of these drugs was determined by clonogenic assay. Total RNA was extracted from the treated cells and RT-PCR was used to determine the effect of the treatment on the expression of RAR beta and ER alpha. Methylation-sensitive PCR analysis was used to confirm that lack of expression of both genes was due to hypermethylation of their promoter regions. A single nucleotide primer extension assay was also used to quantify the reduction in DNA methylation following drug treatment. RESULTS: Both 5-AZA-CdR and TSA alone showed significant antineoplastic activity. The combination of the two drugs was synergistic with respect to MDA-MB-231 cell kill. 5-AZA-CdR alone weakly activated the expression of both RAR beta and ER alpha. TSA alone only activated RAR beta, but not ER alpha. The combination of these agents appeared to produce a greater activation of both genes. CONCLUSIONS: The interesting interaction between 5-AZA-CdR and TSA in both cell kill and cancer-related gene reactivation provides a rationale for the use of inhibitors of DNA methylation and histone deacetylation in combination for the chemotherapy of breast cancer. PMID- 11488528 TI - Pharmacokinetics of Erwinia asparaginase after intravenous and intramuscular administration. AB - PURPOSE: To describe the pharmacokinetics of Erwinia asparaginase (ASNase) after intravenous (i.v.) and intramuscular (i.m.) administration. METHODS: A group of 29 children with newly diagnosed acute lymphoblastic leukemia (ALL) received Erwinia ASNase 30,000 IU/m2 every day for 10 days during multiagent induction therapy. Of these patients. 13 received i.v. therapy and 16 received i.m. therapy. During the reinduction phase the patients received Erwinia ASNase 30,000 IU/m2 twice a week for 2 weeks (Mondays and Thursdays) (8 patients in the i.v. treated group and 11 patients in the i.m.-treated group). ASNase activity (spectrophotometric assay) was measured in plasma samples obtained from the patients at various times during therapy. RESULTS: The estimated half-life was 6.4 +/- 0.5 h (n = 13), the absorption rate after i.m. administration was found to limit elimination. The apparent volume of distribution corresponded well with the volume of plasma. The estimated clearance suggested that Erwinia ASNase is a low-clearance drug. Bioavailability after i.m. administration was (mean +/- SEM) 27.0 +/- 4.5% (range 11-61%; n = 12). CONCLUSIONS: In this study the pharmacokinetic parameters after i.v. and i.m. administration of Erwinia ASNase were determined based on a substantial number of patients. The present findings emphasize the importance of conducting proper pharmacokinetic studies before a new drug or a new preparation of a drug is introduced in a different schedule. PMID- 11488529 TI - Therapeutic activity of 7-[(2-trimethylsilyl)ethyl)]-20 (S)-camptothecin against central nervous system tumor-derived xenografts in athymic mice. AB - PURPOSE: Camptothecins have emerged as an important new class of antitumor drugs. Camptothecin derivatives such as CPT-11 and topotecan are commercially available and approved for the treatment of colorectal (CPT-11) and ovarian and small-cell lung cancer (topotecan). This study was designed to test the efficacy of karenitecin, a novel highly lipophilic camptothecin derivative, against a panel of human tumor xenografts derived from adult and pediatric central nervous system malignancies growing in athymic nude mice. METHODS: Xenografts evaluated were derived from childhood high-grade gliomas (D-212 MG, D-456 MG), adult high-grade gliomas (D-54 MG, D-245 MG), medulloblastomas (D-341 MED, D-487 MED), and ependymomas (D-528 EP, D-612 EP), as well as sublines with demonstrated resistance to procarbazine (D-245 MG (PR)) and busulfan (D-456 (BR)). In replicate experiments, karenitecin was given at 1.0 mg/kg per dose via intraperitoneal injection for a period of 10 consecutive days, which is the dosage lethal to 10% of treated animals. RESULTS: Karenitecin produced statistically significant (P < or = 0.001) growth delays in all subcutaneous xenografts tested, including the sublines resistant to procarbazine and busulfan. Growth delays ranged from 12.1 days in D-456 MG (BR) to 90+ days in D-212 MG and D-341 MED. Karenitecin also produced statistically significant (P < or = 0.001) increases in survival of animals bearing D-341 MED intracranial xenografts (69% increase) and those bearing D-456 MG xenografts (62% increase). CONCLUSION: These preclinical studies confirm that karenitecin is active against human central nervous system xenografts and should undergo clinical evaluation in patients with malignant central nervous system tumors. PMID- 11488530 TI - Combination chemotherapy with ifosfamide, 5-fluorouracil, etoposide and cisplatin for metastatic urothelial cancer. AB - PURPOSE: To investigate the activity of combination chemotherapy with ifosfamide, 5-fluorouracil, etoposide and cisplatin in patients with metastatic urothelial cancer. METHODS: A group of 29 patients were treated with 2000 mg/m2 ifosfamide, 750 mg/m2 5-fluorouracil, 100 mg/m2 etoposide and 20 mg/m2 cisplatin. All four drugs were given intravenously on days 1 through 3 and the treatment was repeated every 3 weeks. Of the 29 patients, 14 had lymph node metastasis alone, and 15 had visceral lesions. RESULTS: An objective response was achieved in 17 patients (59%). There was no difference in response rates according to metastatic site including osseous lesions, which responded well in four of six patients. The 3 year survival rate for all patients was 16% with four patients who had undergone salvage surgery being alive with no evidence of disease 15 to 61 months after initiation of the treatment. A good performance status, lymph node metastasis alone and administration of chemotherapy at the full dosage had a significantly favorable impact on patient survival. Bone marrow toxicity was significant and one patient died of treatment-related sepsis. CONCLUSIONS: Ifosfamide, 5 fluorouracil, etoposide and cisplatin combination chemotherapy appeared to be active in the treatment of metastatic urothelial cancer. Although bone marrow toxicity was significant, the treatment was well tolerated by the majority of the patients. Further study may be warranted. PMID- 11488531 TI - Plasmid expression of a peptide that selectively blocks oncogenic ras-p21-induced oocyte maturation. AB - PURPOSE: We have previously found that a synthetic peptide corresponding to ras p21 residues 96 110 (PNC2) selectively blocks oncogenic (Val 12-containing) ras p21 protein-induced oocyte maturation. With a view to introducing this peptide into ras-transformed human cells to inhibit their proliferation, we synthesized an inducible plasmid that expressed this peptide sequence. Our purpose was to test this expression system in oocytes to determine if it was capable of causing selective inhibition of oncogenic ras-p21. METHODS: We injected this plasmid and a plasmid expressing a control peptide into oocytes either together with oncogenic p21 or in the presence of insulin (that induces maturation that is dependent on normal cellular ras-p21) in the presence and absence of the inducer isopropylthioglucose (IPTG). RESULTS: Microinjection of this plasmid into oocytes together with Val 12-p21 resulted in complete inhibition of maturation in the presence of inducer. Another plasmid encoding the sequence for the unrelated control peptide, X13, was unable to inhibit Val 12-p21-induced maturation. In contrast, PNC2 plasmid had no effect on the ability of insulin-activated normal cellular or wild-type ras-p21 to induce oocyte maturation, suggesting that it is selective for blocking the mitogenic effects of oncogenic (Val 12) ras p21. CONCLUSION: We conclude that the PNC2 plasmid selectively inhibits oncogenic ras p21 and may therefore be highly effective in blocking proliferation of ras induced cancer cells. Also, from the patterns of inhibition, by PNC2 and other ras- and raf-related peptides, of raf- and constitutively activated MEK-induced maturation, we conclude that PNC2 peptide inhibits oncogenic ras p21 downstream of raf. PMID- 11488532 TI - Tissue-type plasminogen activator is involved in the process of neuronal death induced by oxygen-glucose deprivation in culture. AB - Effect of tissue-type plasminogen activator (tPA) on oxygen-glucose deprivation (OGD) was studied in cultured cortical neurons prepared from tPA gene knockout (tPA-KO) and wild-type (Wt) mice. Three hours of OGD induced 45% and 23% of neuronal death in Wt and tPA-KO mice, respectively. Neuronal death in tPA-KO mice was increased to 42% by additional tPA. Six hours of OGD induced 80% and 40% of neuronal death in Wt and tPA-KO mice, respectively, whereas the addition of tPA increased to 62% in tPA-KO mice. These results suggest that tPA is directly involved in the process of neuronal death induced by ischemia-mimic stress without involving vascular or circulatory components. PMID- 11488533 TI - Positron emission tomography compartmental models. AB - The current article presents theory for compartmental models used in positron emission tomography (PET). Both plasma input models and reference tissue input models are considered. General theory is derived and the systems are characterized in terms of their impulse response functions. The theory shows that the macro parameters of the system may be determined simply from the coefficients of the impulse response functions. These results are discussed in the context of radioligand binding studies. It is shown that binding potential is simply related to the integral of the impulse response functions for all plasma and reference tissue input models currently used in PET. This article also introduces a general compartmental description for the behavior of the tracer in blood, which then allows for the blood volume-induced bias in reference tissue input models to be assessed. PMID- 11488534 TI - In vivo measurements of brain glucose transport using the reversible Michaelis Menten model and simultaneous measurements of cerebral blood flow changes during hypoglycemia. AB - Glucose is the major substrate that sustains normal brain function. When the brain glucose concentration approaches zero, glucose transport across the blood brain barrier becomes rate limiting for metabolism during, for example, increased metabolic activity and hypoglycemia. Steady-state brain glucose concentrations in alpha-chloralose anesthetized rats were measured noninvasively as a function of plasma glucose. The relation between brain and plasma glucose was linear at 4.5 to 30 mmol/L plasma glucose, which is consistent with the reversible Michaelis Menten model. When the model was fitted to the brain glucose measurements, the apparent Michaelis-Menten constant, Kt, was 3.3 +/- 1.0 mmol/L, and the ratio of the maximal transport rate relative to CMRglc, Tmax/CMRglc, was 2.7 +/- 0.1. This Kt is comparable to the authors' previous human data, suggesting that glucose transport kinetics in humans and rats are similar. Cerebral blood flow (CBF) was simultaneously assessed and constant above 2 mmol/L plasma glucose at 73 +/- 6 mL 100 g(-1) min(-1). Extrapolation of the reversible Michaelis-Menten model to hypoglycemia correctly predicted the plasma glucose concentration (2.1 +/- 0.6 mmol/L) at which brain glucose concentrations approached zero. At this point, CBF increased sharply by 57% +/- 22%, suggesting that brain glucose concentration is the signal that triggers defense mechanisms aimed at improving glucose delivery to the brain during hypoglycemia. PMID- 11488535 TI - Interdependence of regional and global cerebral blood flow during visual stimulation: an O-15-butanol positron emission tomography study. AB - The authors investigated the influence of variations in global cerebral blood flow (gCBF) on regional flow changes during visual stimulation. Global flow was varied using different end-expiratory CO2 values (PETCO2) between 20 and 70 mm Hg. Visual stimulation was performed with a red LED-array flashing at 8 Hz. Blood flow was measured with 0-15-butanol, continuous arterial blood sampling, and positron emission tomography (PET). Global flow changes surpassed the published values of O-15-H2O studies, better fitting the results of the inert gas technique (gCBF at 20, 40, and 70 mm Hg PETCO2 +/- SD was 31 +/- 4, 48 +/- 13, and 160 +/- 50 mL 100 g(-1) min(-1), respectively). The relation between PETCO2 and CBF in the current study was best described by an exponential rather than a linear function. At low PETCO2, the activation-induced flow changes are moderately damped, whereas at high PETCO2, they are nearly lost (deltaCBF (+/-SD): 52% +/- 25%, 68% +/- 22%, 16% +/- 25% at PETCO2 = 20, 40, 70 mm Hg, respectively). PMID- 11488536 TI - The intraparietal cortex: subregions involved in fixation, saccades, and in the visual and somatosensory guidance of reaching. AB - The functional activity of the intraparietal cortex was mapped with the [14C]deoxyglucose method in monkeys performing fixation of a central visual target, saccades to visual targets, reaching in the light during fixation of a central visual target, and acoustically triggered reaching in the dark while the eyes maintained a straight ahead direction. Different subregions of the intraparietal cortical area 7 were activated by fixation, saccades to visual targets, and acoustically triggered reaching in the dark. Subregions in the ventral part of the intraparietal cortex (around the fundus of the intraparietal sulcus) were activated only during reaching in the light, in which case visual information was available to guide the moving forelimb. In contrast, subregions in the dorsal part of the intraparietal cortical area 5 were activated during both reaching in the light and the dark, in which cases somatosensory information was the only one available in common. Thus, visual guidance of reaching is associated with the ventral intraparietal cortex, whereas somatosensory guidance, based on proprioceptive information about the current forelimb position, is associated with dorsal intraparietal area 5. PMID- 11488537 TI - Neuroprotection in ischemia-reperfusion injury: an antiinflammatory approach using a novel broad-spectrum chemokine inhibitor. AB - Cerebral ischemia-reperfusion injury is associated with a developing inflammatory response with pathologic contributions from vascular leukocytes and endogenous microglia. Signaling chemokines orchestrate the communication between the different inflammatory cell types and the damaged tissue leading to cellular chemotaxis and lesion occupation. Several therapies aimed at preventing this inflammatory response have demonstrated neuroprotective efficacy in experimental models of stroke, but to date, few investigators have used the chemokines as potential therapeutic targets. In the current study, the authors investigate the neuroprotective action of NR58-3.14.3, a novel broad-spectrum inhibitor of chemokine function (both CXC and CC types), in a rat model of cerebral ischemia reperfusion injury. Rats were subjected to 90 minutes of focal ischemia by the filament method followed by 72 hours of reperfusion. Both the lesion volume, measured by serial magnetic resonance imaging, and the neurologic function were assessed daily. Intravenous NR58-3.14.3 was administered, 2 mg/kg bolus followed by 0.5 mg/kg hour constant infusion for the entire 72-hour period. At 72 hours, the cerebral leukocytic infiltrate, tumor necrosis factor-alpha (TNF-alpha), and interleukin-8 (IL-8)-like cytokines were analyzed by quantitative immunofluorescence. NR58-3.14.3 significantly reduced the lesion volume by up to 50% at 24, 48, and 72 hours post-middle cerebral artery occlusion, which was associated with a marked functional improvement to 48 hours. In NR58-3.14.3 treated rats, the number of infiltrating granulocytes and macrophages within perilesional regions were reduced, but there were no detectable differences in inflammatory cell numbers within core ischemic areas. The authors reported increased expression of the cytokines, TNF-alpha, and IL-8-like cytokines within the ischemic lesion, but no differences between the NR58-3.14.3-treated rats and controls were reported. Although chemokines can have pro- or antiinflammatory action, these data suggest the overall effect of chemokine up-regulation and expression in ischemia-reperfusion injury is detrimental to outcome. PMID- 11488538 TI - Induction of Tie-1 and Tie-2 receptor protein expression after cerebral ischemia reperfusion. AB - Tie-1 and Tie-2 are receptor tyrosine kinases (RTKs) that are exclusively expressed in endothelial cells and play important roles in endothelial cell biology. The authors have reported previously the temporal profiles of Tie-1 and Tie-2 mRNA expression after focal cerebral ischemia-reperfusion. In the current study, the localization of Tie-1/Tie-2 mRNA and proteins were further investigated in the same focal ischemia model. In situ hybridization showed that, after 60-minute ischemia and 72-hour reperfusion, both Tie-1 and Tie-2 mRNA appeared as capillary-like structures in the ischemic middle cerebral artery (MCA) cortex. Western blot analysis showed a biphasic expression of Tie-1 protein in the same region. The first peak, spanning the ischemic and early reperfusion period. was of low intensity and short-lived. The second peak was of greater intensity and spanning the period from 72 to 168 hours after reperfusion. Similarly, Tie-2 expression at the protein level also exhibited a biphasic pattern. Immunohistochemical studies, after 72 hours of reperfusion, showed that although Tie-1 and Tie-2 were detected within the ischemic cortex, they actually were expressed in different populations of endothelial cells in different regions. In agreement with the in situ hybridization study, Tie-1 immunoreactivity appeared as capillary-like structures in cortical layers 2 to 4. Similar capillary-like appearance of Tie-2 immunoreactivity was noted in the outer cortical layers. In addition, Tie-2 immunoreactivity also was observed in cortical layer 6b, where de novo large vessel formation was noted. Cellular colocalization experiments revealed that Tie-2 is expressed in proximity to its antagonist, Angpo-2, as well as basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in cortical layer 1, where active vessel remodeling was noted. Interestingly, bFGF only partially colocalized with VEGF, suggesting differential roles for these angiogenic factors during vessel remodeling. Tie-1 protein, to a lesser degree, also colocalized with Angpo-2, bFGF, and VEGF in cortical layer 1. Magnetic resonance imaging (MRI) showed increased regional cerebral blood flow (CBF) corresponding to the expression of these angiogenesis gene products. Together, these findings suggest that the evolving expression of angiogenesis genes underlie the robust vascular remodeling after ischemia and reperfusion. PMID- 11488539 TI - Amyloid beta peptide-induced cerebral endothelial cell death involves mitochondrial dysfunction and caspase activation. AB - Amyloid beta peptide (A beta), a 39 to 43 amino acid fragment of the beta-amyloid precursor protein (betaAPP), forms insoluble fibrillar accumulation in neurofibrillary tangles and vascular plaques. A beta has been implicated in neuronal and vascular degeneration in brain regions susceptible to plaque formation because of its cytotoxic effect on neurons and endothelial cells (ECs). The authors used a murine cerebral endothelial cell (CEC) line and primary cultures of bovine CECs to explore the cytotoxic mechanism of A beta. A beta 1-40 and A beta 25-35 peptides caused cell death in a dose-dependent and time dependent manner. Exposure to either A beta 25-35 or A beta 1-40 at 10 micromol/L for 48 hours caused at least 40% cell death. Cerebral endothelial cell death was characterized by nuclear condensation, mitochondrial dysfunction, and nuclear and mitochondrial DNA damage. A beta 25-35 activated both caspase-8 and caspase-3 in murine CECs. zVAD-fmk, a broad-spectrum caspase inhibitor, prevented A beta 25-35 induced increase in caspase-3 activity and CEC death. N-acetyl-cysteine, an antioxidant, also prevented A beta-induced cell death. Together, these findings indicate that A beta-mediated CEC death is an apoptotic process that is characterized by increased oxidative stress, caspase activation, mitochondrial dysfunction, and nuclear and mitochondrial DNA damage. PMID- 11488540 TI - Na+-K+-Cl- cotransporter in rat focal cerebral ischemia. AB - In cultured neurons, the authors previously demonstrated that the Na+-K+-Cl- cotransporter is significantly stimulated by elevated extracellular potassium and glutamate, which are important factors in cerebral ischemic damage. These findings led the authors to hypothesize that stimulation of the cotransporter after ischemia might result in Na+, K+, and Cl- influx, and might contribute to neuron damage. In the current study, the authors investigated such a role of the Na+-K+-Cl- cotransporter in focal cerebral ischemia. Cerebral ischemia was induced by 2-hour occlusion of the left middle cerebral artery (MCA) and 24-hour reperfusion in male spontaneously hypertensive rats (SHRs). Immunocytochemical staining and immunoblotting revealed an up-regulation of expression of the cotransporter protein in neurons in cortex at 24 hours of reperfusion. Artificial cerebral spinal fluid (aCSF) or 100 micromol/L bumetanide (a cotransporter inhibitor) in aCSF were continuously microdialyzed through a microdialysis probe into left cortices throughout 2-hour MCA occlusion and 24-hour reperfusion. Compared with the aCSF-treated group, infarction volume was significantly reduced in the bumetanide-treated group (25%, P < 0.05). In addition, brain water content in the bumetanide-treated brains was decreased by 70% (P < 0.05). These results strongly suggest that the Na+-K+-Cl- cotransporter may play an important role in cerebral ischemic neuronal damage. PMID- 11488541 TI - Attenuation of ischemia-induced mouse brain injury by SAG, a redox-inducible antioxidant protein. AB - Cerebral ischemia resulting from a disruption of blood flow to the brain initiates a cascade of events that causes neuron death and leads to neurologic dysfunction. Reactive oxygen species are thought, at least in part, to mediate this disease process. The authors recently cloned and characterized an antioxidant protein, SAG (sensitive to apoptosis gene), that is redox inducible and protects cells from apoptosis induced by redox agents in a number of in vitro cell model systems. This study reports a neuroprotective role of SAG in ischemia/reperfusion-induced brain injury in an in vivo mouse model. SAG was expressed at a low level in brain tissue and was inducible after middle cerebral artery occlusion with peak expression at 6 to 12 hours. At the cellular level, SAG was mainly expressed in the cytoplasm of neurons and astrocytes, revealed by double immunofluorescence. An injection of recombinant adenoviral vector carrying human SAG into mouse brain produced an overexpression of SAG protein in the injected areas. Transduction of AdCMVSAG (wild-type), but not AdCMVmSAG (mutant), nor the AdCMVlacZ control, protected brain cells from ischemic brain injury, as evidenced by significant reduction of the infarct areas where SAG was highly expressed. The result suggests a rather specific protective role of SAG in the current in vivo model. Mechanistically, SAG overexpression decreased reactive oxygen species production and reduced the number of apoptotic cells in the ischemic areas. Thus, antioxidant SAG appears to protect against reactive oxygen species-induced brain damage in mice. Identification of SAG as a neuroprotective molecule could lead to potential stroke therapies. PMID- 11488542 TI - Global ischemia induces expression of acid-sensing ion channel 2a in rat brain. AB - Acid-sensing ion channels (ASICs) are ligand-gated cation channels that respond to acidic stimuli. They are expressed throughout the mammalian nervous system. In the peripheral nervous system, ASICs act as nociceptors, responding to the tissue acidosis that accompanies ischemic and inflammatory conditions. The function of ASICs in the central nervous system is not known. In this article, the authors present evidence that transient global ischemia induces ASIC 2a protein expression in neurons that survive ischemia. Western blot analysis with an anti ASIC 2a antibody revealed up-regulation of an 80 kD protein in ischemic rat brain. Immunohistochemical analysis showed that ASIC 2a protein expression increased in neurons of the hippocampus and cortex. Klenow fragment-mediated labeling of DNA strand breaks determined that ASIC 2a induction did not occur in cells with detectable DNA damage. The current results suggest a possible role for ASICs in mediating a cellular response to ischemia. PMID- 11488543 TI - Fractal branching pattern in the pial vasculature in the cat. AB - Arborization pattern was studied in pial vascular networks by treating them as fractals. Rather than applying elaborate taxonomy assembled from measures from individual vessel segments and bifurcations arranged in their branching order, the authors' approach captured the structural details at once in high-resolution digital images processed for the skeleton of the networks. The pial networks appear random and at the same time having structural elements similar to each other when viewed at different scales--a property known as self-similarity revealed by the geometry of fractals. Fractal (capacity) dimension, Dcap, was calculated to evaluate the network's spatial complexity by the box counting method (BCM) and its variant, the extended counting method (XCM). Box counting method and XCM were subject to numerical testing on ideal fractals of known D. The authors found that precision of these fractal methods depends on the fractal character (branching, nonbranching) of the structure they evaluate. Dcaps (group mean +/- SD) for the arterial and venous pial networks in the cat (n = 6) are 1.37 +/- 0.04, 1.37 +/- 0.02 by XCM, and 1.30 +/- 0.04, 1.31 +/- 0.03 by BCM, respectively. The arterial and venous systems thus appear to be developed according to the same fractal generation rule in the cat. PMID- 11488544 TI - Respiratory tract toxicity in rats exposed to Mexico City air. AB - The rat has been used extensively as a health sentinel, indicator, or monitor of environmental health hazards, but this model has not been directly validated against human exposures. Humans in Mexico City show upper respiratory tract lesions and evidence of pulmonary damage related to their environmental inhalation exposure. In this study, male and female F344 rats were exposed (23 hr/day) in Mexico City to local Mexico City air (MCA)* for up to seven weeks. Controls were maintained at the same location under filtered air. Prior to these exposures, several steps were taken. First, the nasal passages of normal male rats shipped from the United States and housed in Mexico City were examined for mycoplasma infection; no evidence of infection was found. In addition, a mobile exposure and monitoring system was assembled and, with an ozone (O3) exposure atmosphere, was tested along with supporting histopathology techniques and analysis of rat nasal and lung tissues. Last, the entire exposure model (equipment and animals) was transported to Mexico City and validated for a three week period. During the seven-week study there were 18 one-hour intervals during which the average O3 concentration of MCA in the exposure chamber exceeded the US National Ambient Air Quality Standard (NAAQS) of 0.120 ppm 03 (hourly average, not to be exceeded more than once per year). This prolonged exposure of healthy F344 rats to MCA containing episodically low to moderate concentrations of 03 (as well as other urban air pollutants) did not induce inflammatory or epithelial lesions in the nasal airways or lung as measured by qualitative histologic techniques or quantitative morphometric techniques. These findings agree with those of previous controlled O3 inhalation studies, but they are in contrast to reports indicating that O3-polluted MCA causes significant nasal mucosal injury in adults and children living in southwestern Mexico City. Taken together, these findings may suggest that human airways are markedly more susceptible to the toxic effects of MCA than are the airways of the F344 rat. PMID- 11488545 TI - Respiratory epithelial penetration and clearance of particle-borne benzo[a]pyrene. AB - Exposure to diesel exhaust is a suspected risk factor for human lung cancer. The carbonaceous core of the soot particles found in diesel exhaust and the condensed organic compounds adsorbed (or bound) onto the surface of the particles are both possible contributors to this suspected risk. The extent and rate at which organic procarcinogens desorb from soot particles in the lungs after environmental and workplace exposures and the degree of metabolic activation in the lungs are also not known. We explored the relationship between a model polynuclear aromatic hydrocarbon (PAH)* and a typical carrier particle by measuring the rate of release, extent of release, and metabolic fate of benzo[a]pyrene (BaP) bound onto the carbonaceous core of diesel soot after bolus aerosol exposures of the dog's peripheral lung and trachea. Exogenous BaP was bound onto preextracted diesel soot at a surface concentration corresponding to 25% of a monomolecular layer. After deposition in the alveolar region, a fraction of BaP was rapidly desorbed from the soot and quickly absorbed into the circulating blood. Release rates then decreased drastically. When the BaP coating reached approximately 16% of a monolayer, it was not bioavailable and remained on the particles after 5.6 months in the lung. The bioavailability of BaP on particles retained in lymph nodes was markedly higher, however: after 5.6 months the surface coating of BaP was reduced to 10% of a monolayer. Fractions of BaP that remained bound to the soot surface during this 5.6 months had a low reactivity-nearly 30% of the radioactive compounds extracted from recovered soot particles were still BaP, the parent compound. In contrast, the rapidly released fraction of BaP, which was quickly absorbed through the alveolar epithelium after inhalation, appeared mostly unmetabolized in the circulation, along with low concentrations of phase I and phase II BaP metabolites. Within approximately 1 hour, however, this rapidly absorbed fraction of BaP was metabolized, most likely in the liver, with the metabolite spectrum being dominated by conjugated phase II metabolites. The fraction of BaP desorbed from particles deposited on the epithelium of the conducting airways was absorbed by the epithelium but slowly penetrated the capillary bed. The absorbed BaP was rapidly metabolized in the airway epithelium, as indicated by the influx of tritiated water (3H2O) from the lungs into the circulation. The results suggest that the dosimetry of inhaled, highly lipophilic BaP during typical exposures is bimodal. The larger fraction of bioavailable BaP deposited in the alveolar region was absorbed mostly unaltered into the blood through the alveolar type I cells and was metabolized systemically. A smaller fraction of bioavailable BaP was deposited on the airway mucosa and rapidly metabolized, most likely in the airway epithelium. The substrate levels of BaP in the epithelium of the conducting airways exceeded the systemic levels by up to two orders of magnitude. This dramatic site-of-entry to systemic duality in the dosimetry of inhaled BaP is likely to be similar in most mammalian species and should be considered in risk assessment models for PAHs in humans. PMID- 11488546 TI - Relationship between life satisfaction and violent behaviors among adolescents. AB - OBJECTIVE: To explore relationships between life satisfaction and violent behaviors among 5,032 adolescents. METHODS: Adjusted polychotomous logistic regression analyses and multivariate models were used via SUDAAN with the 1997 CDC YRBS. RESULTS: Carrying a weapon; carrying a gun; carrying a weapon at school; physical fighting; physical fighting at school; physical fighting that required physician treatment; drinking and driving; riding with a drinking driver; having property stolen/damaged at school; feeling unsafe while at, going to or returning from school; and being injured/ threatened with a weapon were associated (p=.05) with reduced life satisfaction. CONCLUSION: Measures of life satisfaction as a component of comprehensive assessments of adolescent violence/aggression in field-work, research, and program-evaluation efforts should be considered. PMID- 11488547 TI - Assets as predictors of suicide attempts in African American inner-city youths. AB - OBJECTIVE: To assess whether developmental assets and risk-taking behaviors, as measured by the Search Institute instrument, were related to attempted suicide in African American, inner-city youth. METHODS: Survey of 336 African American, inner-city youths. RESULTS: Significant odds ratios were found for 4 asset questions and for 11 risk-taking behavior questions by attempted suicide behavior. The odds ratios ranged from 2.4 to 6.4 in magnitude. The percent of variation in suicide-attempt behavior explained by the asset questions (14.6% and 19.6%) was explained by the risk-taking behavior/high-risk behavior pattern questions. CONCLUSIONS: The risk-taking behavior items were better predictors of suicide behavior than were the developmental asset items for the African American youth. PMID- 11488548 TI - A comparison of teen perceptions and parental reports of influence on driving risk. AB - OBJECTIVES: To compare associations between teen and parent reports of parental driving influence to teen-reported high-risk driving. METHODS: A statewide sample of 424 Maryland parents and their provisionally licensed teenagers were interviewed. RESULTS: Reports of parental involvement with their driving to that of their parents indicated low levels of concordance. Teens who reported their parents had specific rules restricting who could ride with them as well as how many passengers were allowed to ride with them were less likely to report being distracted by friends, getting a ticket, driving too fast, or driving aggressively. CONCLUSION: Programs that enable parents to restrict and supervise their teen's driving, especially during the first 6 months of provisional licensure, are indicated. PMID- 11488549 TI - The role of optimism in health-enhancing behavior. AB - OBJECTIVE: To examine the relationship between levels of optimism and participation in health-enhancing behaviors. METHODS: A convenience sample of volunteers obtained from undergraduate psychology classes (N=118) completed self report measures of optimism and current health habits. They also predicted their use of various health strategies should they be diagnosed with a life-threatening illness. RESULTS: Analyses revealed a positive association between scores on the measure of optimism and both of the health measures. CONCLUSIONS: The study extended earlier findings of a positive relationship between optimism and health enhancing behaviors and demonstrated that this relationship can be observed for general health habits as well as in the context of a specific health threat. PMID- 11488550 TI - Evaluation of a culturally appropriate intervention to increase physical activity. AB - OBJECTIVE: To evaluate a culturally appropriate intervention to increase activity in overweight Mexican American women. METHODS: Participants were randomly assigned to a physical activity program or wait-list control. RESULTS: Treated participants were not more active than controls at 6 or 12 months. In addition, we found no significant differences in the proportion of individuals who met an objective criterion for physical activity from baseline to 6 months in the treatment or control groups. CONCLUSION: The intervention did not increase physical activity in this population. Differences in baseline activity and contamination of the control group may partially account for the outcome. PMID- 11488551 TI - Internal associations among health-risk factors and risk prevalence. AB - OBJECTIVE: To explore risk associations between health variables and to compare these associations with the prevalence of each risk. METHODS: Manufacturing company employees (N=16,879) completed a self-reported health risk appraisal with on-site biometric screening. RESULTS: Risk prevalence and risk association findings differed by gender, age, and overall risk level. Risks that were most highly associated with other risks were different from risks that were most prevalent. CONCLUSIONS: The findings suggest that in addition to risk prevalence, individual characteristics and the level of association between risks are also important to consider when planning health interventions. PMID- 11488552 TI - Social environmental influences on adolescents' smoking progression. AB - OBJECTIVE: To determine social environmental factors in relation to adolescent smoking using both longitudinal and cross-sectional analyses. METHODS: A national sample of nonsmoking adolescents (N=4,431) who participated in the 1989 Teenage Attitudes and Practices Survey and who were re-interviewed in 1993 was used for this study. RESULTS: The smoking behavior of best friends and smoking beliefs were consistent and significant factors in predicting adolescents' smoking progression from nonsmoking status to regular smokers or experimental smokers. CONCLUSIONS: By addressing several limitations of previous studies, this paper contributes additional information regarding the adolescents' smoking acquisition process. PMID- 11488553 TI - Perinatal lead exposure alters the expression of neuronal nitric oxide synthase in rat brain. AB - Environmental exposure to lead (Pb) is known to affect the developing nervous system causing cognitive deficits in children. The diffusible nitric oxide (NO) is a biological messenger known to be involved in brain development. We examined the developmental changes of neuronal nitric oxide synthase (nNOS) in cerebellum and hippocampus of developing rat brain by radiometric assay, Western blot analysis and immunohistochemistry. Pb-exposure (0.2% Pb acetate) was initiated on gestation day 6 through the drinking water of the dam and continued through birth and postnatal days (PNDs) 1 to 21. The pups were never exposed to Pb directly. Pb exposure was stopped on weaning of pups from mothers on PND 21. The changes in nNOS were measured in the offspring on PNDs 7, 14, 21, and 35. The nNOS activity was increased gradually from PNDs 7 to 35 in both cerebellum and hippocampus of control rats when the enzyme activity was determined in the presence of either 0.5 or 6 microM calcium (Ca2+) in the reaction mixture. However, Pb exposure decreased the nNOS activity significantly at PNDs 21 to 35 as compared to respective controls when the enzyme activity was determined in the presence of 6 microM Ca2+. The decrease of nNOS was even greater and evident at all PNDs tested when the enzyme activity was assayed in the presence of physiological concentration of Ca2+ (0.5 microM). These findings were further strengthened by the in vitro studies. The cerebellar nNOS activity was inhibited much more at low Ca2+ (0.5 microM) as compared to 6 microM Ca2+, with IC50 values of 35 and 50 nM Pb, respectively. The nNOS protein levels and immunoreactivity in the cerebellum and hippocampus of rats perinatally exposed to Pb were decreased as compared to controls at PNDs 21 and 35. These data suggest perinatal Pb exposure decreases the nNOS in the developing brain. The decrease of nNOS activity and protein may explain the Pb-mediated cognitive deficits because NO regulates long-term potentiation (LTP) and other neurophysiological events in the developing nervous system. PMID- 11488554 TI - Two-generation reproduction study and immunotoxicity screen in rats dosed with phenol via the drinking water. AB - This study evaluated the potential reproductive toxicity of phenol in a rat two generation reproduction study, which included additional study endpoints, such as sperm count and motility, developmental landmarks, histological evaluation of suspect target organs (liver, kidneys, spleen, and thymus), weanling reproductive organ weights, and an immunotoxicity screening plaque assay. Phenol was administered to 30 Sprague-Dawley rats/sex/group in the drinking water at concentrations of 0, 200, 1000, or 5000 ppm. Parental (P1) animals were treated for 10 weeks prior to mating, during mating, gestation, lactation, and until sacrifice. The F1 generation (P1 offspring) was treated using a similar regimen, while the F2 generation was not treated. After mating, 10 P1 males/group were evaluated using standard clinical pathology parameters and an immunotoxicity screening plaque assay. Significant reductions in water and food consumption were observed in the 5000-ppm group in both generations; corollary reductions in body weight/body weight gain were also observed. Mating performance and fertility in both generations were similar to controls, and no adverse effects on vaginal cytology or male reproductive function were observed. Vaginal opening and preputial separation were delayed in the 5000-ppm group, and were considered to be secondary to the reduction in F1 body weight. Litter survival of both generations was reduced in the 5000-ppm group. Absolute uterus and prostate weights were decreased in the F1 generation at all dose levels; however, no underlying pathology was observed and there was no functional deficit in reproductive performance. Therefore, these findings were not considered to be adverse. No evidence of immunotoxicity was noted in the 5000-ppm group. The effects noted at the high concentration were presumed to be associated with flavor aversion to phenol in the drinking water. Based on a comprehensive examination of all parameters, the no-observable-adverse-effect level (NOAEL) for reproductive toxicity of phenol administered in drinking water to rats is 1000 ppm. The corresponding daily intake of phenol for an adult rat at the NOAEL of 1000 ppm is equivalent to about 70 mg/kg/day for males and 93 mg/kg/day for females. PMID- 11488555 TI - Introduction/overview: gender-based differences in pharmacologic and toxicologic responses. AB - Gender may be the most important factor in mammalian development and response to exogenous agents. From believing sex-related differences required sheltering women to protect their reproductive capacity (Victorians thought exercise, education, train travel, and certain music neuro- and reprotoxic to females) to legislating a status of essential equality of the sexes may have increased women's health issues. Men and women often respond differently to drugs. Inclusion of women in phase I/II clinical trials is insufficient to identify gender-based differences in response; rather, animal models should be the basis for predicting gender-based differences in pharmacologic and toxicologic effects. Unfortunately, current animal models do not consistently demonstrate such differences. Use of commonly used species (e.g., rats and dogs) does not necessarily result in relevant evaluation of an agent in a species at appropriate development (age), physiological state, anatomy, metabolism, or kinetics for estimation of human risks. The need to test agents in relevant animal models and advances in metabolic, pharmacokinetic, and pharmacodynamic capabilities challenge us to improve methods by using the most relevant models for estimating human risk. We need to be concerned about gender-related differences and the dynamics of gender-based growth and development over the entire life cycle. We must also consider potential interactions of dietary supplements and other exogenous agents that can act as drugs or modulate the potential effects of drugs differently in men, women, and developing children of both sexes. To this end, the health benefits of genistein and the effects of this dietary agent in a multigeneration study in rats will be described. It is envisioned that this symposium will assist in re-recognition of the importance of gender-related differences in use and response to pharmaceuticals and result in optimization of nonclinical testing procedures to identify benefits and risks for human use of these agents. PMID- 11488556 TI - Gender-based differences in the toxicity of pharmaceuticals--the Food and Drug Administration's perspective. AB - Women experience more adverse reactions to treatment with therapeutic drugs than men. Theories proposed to explain this include overdosing, different pharmacokinetics and pharmacodynamics, women are more likely to report adverse events than men, or women take more medications than men. Food and Drug Administration (FDA) Office of Women's Health (OWH) funds research to promote including women in clinical trials and understanding the biology of sex-related differences in the safety of FDA-regulated products. Including women in clinical trials advances the understanding of drug efficacy and safety in women by providing information on drug dosing, pharmacokinetics, and pharmacodynamics. A Baysian statistical analysis of sex differences in adverse events showed that although about the same number of adverse events were reported for men and women, those reported for women were more serious. One example of a sex difference in the toxicity of pharmaceuticals is the drug-induced cardiac arrhythmia, torsades de point. OWH funded studies in animals and humans to investigate the mechanism behind this sex difference. These studies demonstrated that shortening the QT interval increases the risk of developing torsades and that androgens protect against torsades by slowing cardiac repolarization and prolonging the QT interval. Understanding the mechanisms behind other reported sex-related differences in adverse drug effects requires additional research. The preliminary studies conducted to date suggest that this sex-related difference is likely to be a multifactorial problem requiring information from several fields of study. Ideally, individuals at risk for developing an adverse event should be identified prior to therapeutic intervention. The OWH plans to fund more studies to investigate the role of hormonal variations on drug metabolism and drug-drug interactions. Animal and in vitro model systems are needed to fully understand the mechanism of how gender influences drug toxicity. PMID- 11488557 TI - Animal models used in identifying gender-related differences. AB - There is general agreement in the scientific community on the need to identify appropriate animal models that can be used to screen for gender-based differences. At the same time, there is a growing expectation for data from these models to mimic or be more predictive of the human experience. The species in this review will include nonhuman primates, rats, mice, rabbits, swine, hamsters, gerbils, quail, and fish. Although some of the models are unique, the gender related differences, in most instances, may be correlated with man, due to the conservation of biological systems across species. PMID- 11488558 TI - Gender-based differences in pharmacokinetics in laboratory animal models. AB - The study of gender-based differences in the pharmacokinetics (PK) of compounds tested in animal models has received greater attention in recent years. As early as 1932, the pharmacological action of barbiturates was recognized as gender dependent-female rats required half the dose needed by male rats to induce sleep. Later, it was shown that gender differences in hepatic metabolism were responsible for this gender-related pharmacodynamic response. Today, it is well known that gender-dependent metabolism in rats often results from differences in expression of hepatic enzymes. The sex-specific cyctochrome P450s CYP2C11, CYP2C13, and CYP3A2 are expressed in males whereas CYP2C12 is expressed in females. Most of the known gender-related differences in toxicity of compounds in rats are due to gender-related hepatic metabolism differences. It is clear that compounds may undergo gender-dependent metabolism; it is also true that the fundamental PK parameters of clearance (CL) and volume (V) can demonstrate a gender dependence in a wide variety of animal species: rats, mice, rabbits, hamsters, dwarf goats, cattle, and rainbow trout. To appreciate how gender related differences affect PK parameters, it is necessary to have a basic understanding of the factors that control the PK of compounds. Changes in these factors will be related to the primary PK parameters of CL and V. A review of the literature provides examples of gender-based differences in these factors and examples of the observed differences in the PK profile of the administered compound. Examples of gender-based differences in the PK of compounds leading to gender differences in the toxicity in nonclinical test species are also discussed. PMID- 11488559 TI - Developmental effects and health aspects of soy protein isolate, casein, and whey in male and female rats. AB - Dietary factors other than the traditional nutrients are found in the so-called functional foods. They are becoming increasingly recognized as potentially important for maintaining good health. Soybeans are rich in such factors thought to help prevent certain chronic diseases. Soy protein isolate (SPI) is one of the three major proteins used in infant formulas sold in the United States, with casein (CAS) and whey (WPH) proteins being the others. We have been studying the health effects of these proteins. Safety concerns have developed over the consumption of soy-based infant formula, partly because of the high circulating levels of the total isoflavones (phytoestrogens) during "critical periods of infant development." There is a paucity of data on developmental, physiological, neurophysiological, behavioral, metabolic, or molecular effects of soy phytochemicals in humans, especially during pregnancy and infancy. We have studied the effects of CAS, SPI, and WPH in short-term, long-term, and multigenerational studies in rats. Aside from minor differences in body weight gain profiles, CAS-, SPI- or WPH-fed rats did not differ in development, organ weights, in vitro hepatic metabolism of testosterone (T), or reproductive performance. However, some endocrine-related functions differed between rats fed these proteins. We found that SPI accelerated puberty in female rats (p < .05) and WPH delayed puberty in males and females, as compared with CAS (p < .05). Gender differences were also found in gonadectomy-induced steroid responses. Male rats had normal serum T levels, but female rats fed SPI had reduced serum 17beta estradiol concentrations and a blunted 17beta-estradiol response to ovariectomy, as compared to rats fed CAS or WHP (p < .05). Female rats fed SPI or WHP or treated with genistein had reduced incidence of chemically induced mammary cancers (p < .05) compared to CAS controls, with WHP reducing tumor incidence by as much as 50%, findings that replicate previous results from our laboratory. Together, these results suggest gender-specific differences in development and certain endocrine responses among rats fed diets composed of a single protein source such as those used in infant formulas. Whether similar developmental effects occur in human infants is unknown, but unlikely because (1) most infants do not consume such diets throughout life as these rats did, and (2) no such effects have been reported in millions of American infants fed infant formula containing these proteins. The long-term health consequence implications of early diet exposure to SPI and WPH, such as reduced breast cancer incidence, are likely to be very positive. PMID- 11488560 TI - Gender-based differences in rats after chronic dietary exposure to genistein. AB - Gender-based differences can be observed from pharmacokinetic, behavioral, or anatomical assessments. No single assessment tool will provide a complete answer, but the use of a variety of indices, each with known gender-related outcome differences, can reveal agent-induced gender-based alterations. In a series of initial range-finding studies in rats conducted at the National Center for Toxicological Research (NCTR), the effects of dietary exposure to the weak estrogen, genistein, have been assessed using a number of techniques with validated gender-related outcome measures. The findings indicated that (1) the internal dose of genistein was higher in females than males after equivalent dietary exposure and this was consistent with the faster rate of genistein elimination in males; (2) in behavioral assessments, males and females in the high-dose dietary genistein group consumed more of a sodium-flavored solution; however, no genistein-related changes were observed in open field or running wheel activity, play behavior, or intake of a saccharin-flavored solution; and (3) dose-related alterations of the volume of the sexually dimorphic nucleus of the medial preoptic area were observed in genistein-exposed male rats but not females. These observations describe the utility of a variety of gender-based assessment tools and indicate that dose-related effects of developmental and chronic dietary exposure to genistein can be observed in the rodent. Additional studies, perhaps in nonhuman primates, are necessary to further predict the effect(s) of genistein on human gender-based development. PMID- 11488561 TI - New radiation therapy rooms should be designed to accomodate treatments of the future such as IMRT and tomotherapy. PMID- 11488562 TI - Description and dosimetric verification of the PEREGRINE Monte Carlo dose calculation system for photon beams incident on a water phantom. AB - PEREGRINE is a three-dimensional Monte Carlo dose calculation system written specifically for radiotherapy. This paper describes the implementation and overall dosimetric accuracy of PEREGRINE physics algorithms, beam model, and beam commissioning procedure. Particle-interaction data, tracking geometries, scoring, variance reduction, and statistical analysis are described. The BEAM code system is used to model the treatment-independent accelerator head, resulting in the identification of primary and scattered photon sources and an electron contaminant source. The magnitude of the electron source is increased to improve agreement with measurements in the buildup region in the largest fields. Published measurements provide an estimate of backscatter on monitor chamber response. Commissioning consists of selecting the electron beam energy, determining the scale factor that defines dose per monitor unit, and describing treatment-dependent beam modifiers. We compare calculations with measurements in a water phantom for open fields, wedges, blocks, and a multileaf collimator for 6 and 18 MV Varian Clinac 2100C photon beams. All calculations are reported as dose per monitor unit. Aside from backscatter estimates, no additional, field-specific normalization is included in comparisons with measurements. Maximum discrepancies were less than either 2% of the maximum dose or 1.2 mm in isodose position for all field sizes and beam modifiers. PMID- 11488563 TI - A variable critical-volume model for normal tissue complication probability. AB - Predicting late-term normal-tissue complication probability (NTCP) after radiotherapy is an important factor in the optimization of conformal radiotherapy. We propose a new NTCP model, based on the properties of the high dose region. The principal assumption of the new model is that a whole-organ complication will occur when the radiation damage to a normal organ volume (a portion of the total organ) exceeds a threshold value. The dose threshold for complications varies with the size of the volume (percent of the total organ). We hypothesize that a complication occurs if the complication threshold is exceeded for any organ volume. We used the average dose to a volume as a measure of radiation damage to that volume. Also, we used the power law to scale the average dose to various organ volumes to a whole-organ equivalent dose, and to identify the volume with the most harmful dose-size combination-the critical volume. We used a logistic distribution to calculate the probability that the patient will develop a complication, given the dose delivered to the critical volume. We used a maximum likelihood fit to estimate the model parameters for late-term rectal complications in a set of patients treated for prostate carcinoma with external photon beam radiotherapy (EBRT). Good correspondence was found between the experimental data and the model predictions. PMID- 11488564 TI - Irregular field calculation on the central beam axis of photon beams using sector integration. AB - A method is proposed for calculation of irregular field factors on the central beam axis and homogeneous medium for x-ray beams. The irregular field factor is introduced as the ratio of the output of a field with and without blocks on the central beam axis. The algorithm is based on the sector-integration method and the circular field quantities are calculated from in-phantom measurements. These circular field quantities are the output per beam monitor unit for circular fields defined by a hypothetical secondary collimator and reduced to a circular field by blocking. A derivation of the sector-integration equation is given from first principles. As it is shown, the circular field quantities are evaluated from data measured for rectangular, block shaped fields. Such quantities contain all beam components, including photons scattered from the blocks, the block tray, and photons scattered in the phantom. Consequently, the so called primary and secondary beam components are readily incorporated in this approach. Once the circular field quantities have been determined from rectangular field data, the irregular field factors for other geometry can be calculated. Irregular field factors for square, rectangular and circular block-shaped fields were calculated for 6 MV photon beams and compared with measured values. The results agree within 0.7%, even for heavy blocked field cases, i.e., a 40 x 40 cm2 collimator field blocked to a 5 x 5 cm2 field. The method was tested for a particular source to surface distance, depth, phantom composition, and source to block distance. Calculation of irregular field factors in another set up conditions requires the measurement of the appropriate input data. PMID- 11488565 TI - Characteristics of bremsstrahlung in electron beams. AB - Clinical electron beams contain an admixture of bremsstrahlung produced in structures in the accelerator head, in field-defining cerrobend or lead cutouts, and in the irradiated patient or water phantom. Accurate knowledge of these components is important for dose calculations and treatment planning. In this study, the bremsstrahlung components are separated for electron beams (energy 6 22 MeV, diameter 0-5 cm) using measurements in water and calculations. The results show that bremsstrahlung from the accelerator head dominates and increases with field size for electron beams generated by accelerators equipped with scattering foils. The bremsstrahlung from the field-defining cerrobend accounts for 10% to 30% of the total bremsstrahlung and decreases with increasing beam radius. The bremsstrahlung is softer than the x-ray beams of corresponding nominal energy since the latter are hardened by the flattening filter. For the 6, 12, and 22 MeV electron beams, the effective attenuation coefficients in water for the bremsstrahlung are 0.058, 0.050, and 0.043 cm(-1). The depths of maximum dose at 100 cm SSD are 0.8, 1.7, and 3.0 cm. The position of the virtual source of the bremsstrahlung shifts downstream from the nominal source position by 20, 13, 5.6 cm, respectively. The lateral bremsstrahlung dose distribution is more forward-peaked for higher electron energy. The bremsstrahlung components could be described for any machine by a set of simple measurements and can be modeled by an analytical expression. PMID- 11488566 TI - Graphical treatment simulation and automated collision detection for conformal and stereotactic radiotherapy treatment planning. AB - The purpose of this work is to develop a "room's eye view" graphical simulation program with an automated collision detection option, to assist a treatment planning user to visualize the treatment setup checking at the same time the feasibility of his plan. The program simulates the treatment process using accurate three-dimensional graphical models of the gantry, table, and that of an average patient. This allows the use of any mechanical movement concerning the radiotherapy unit. The simulation of the mechanical movements and their limitations are according to IEC standards and thus compatible with any radiotherapy machine. Graphical methods for collision detection between the models guarantee speed and accuracy. The module simulates the treatment setup with accuracy better than 2 degrees for any tested case. When a collision takes place a warning message is displayed. In this paper a software tool is developed that can be used as a stand-alone program or embedded in any treatment planning system. The visualization of the treatment fields prior to treatment permits the geometric feasibility of the plan, thus adding one more step toward the automation of the treatment process. PMID- 11488567 TI - Detective quantum efficiency of a direct-detection active matrix flat panel imager at megavoltage energies. AB - The use of an amorphous selenium (a-Se) based direct-detection active matrix flat panel imager (AMFPI) is studied for megavoltage imaging. The detector consists of a 1.2 mm copper front plate and 200 microm a-Se layer, and has a 85 microm pixel pitch. The Modulation Transfer Function (MTF), Noise Power Spectrum (NPS), and Detective Quantum Efficiency (DQE) are measured for 6 and 15 MV photon beams. A theoretical expression for the DQE is derived using a recently developed formalism for nonelementary cascade stages. A comparison of theory with experiment is good for the 6 and 15 MV beams. The model is used to explore the DQE for more typical pixel sizes. The results indicate that with proper modifications, such as a larger a-Se thickness, a direct flat-panel AMFPI is a very promising detector for megavoltage imaging. PMID- 11488568 TI - Dosimetry of beta-ray ophthalmic applicators: comparison of different measurement methods. AB - An international intercomparison of the dosimetry of three beta particle emitting ophthalmic applicators was performed, which involved measurements with radiochromic film, thermoluminescence dosimeters (TLDs), alanine pellets, plastic scintillators, extrapolation ionization chambers, a small fixed-volume ionization chambers, a diode detector and a diamond detector. The sources studied were planar applicators of 90Sr-90Y and 106Ru-106Rh, and a concave applicator of 106Ru 106Rh. Comparisons were made of absolute dosimetry determined at 1 mm from the source surface in water or water-equivalent plastic, and relative dosimetry along and perpendicular to the source axes. The results of the intercomparison indicate that the various methods yield consistent absolute dosimetry results at the level of 10%-14% (one standard deviation) depending on the source. For relative dosimetry along the source axis at depths of 5 mm or less, the agreement was 3% 9% (one standard deviation) depending on the source and the depth. Crucial to the proper interpretation of the measurement results is an accurate knowledge of the detector geometry, i.e., sensitive volume and amount of insensitive covering material. From the results of these measurements, functions which describe the relative dose rate along and perpendicular to the source axes are suggested. PMID- 11488569 TI - Calculation of beta-ray dose distributions from ophthalmic applicators and comparison with measurements in a model eye. AB - Dose distributions throughout the eye, from three types of beta-ray ophthalmic applicators, were calculated using the EGS4, ACCEPT 3.0, and other Monte Carlo codes. The applicators were those for which doses were measured in a recent international intercomparison [Med. Phys. 28, 1373 (2001)], planar applicators of 106Ru-106Rh and 90Sr-90Y and a concave 106Ru-106Rh applicator. The main purpose was to compare the results of the various codes with average experimental values. For the planar applicators, calculated and measured doses on the source axis agreed within the experimental errors (<10%) to a depth of 7 mm for 106Ru-106Rh and 5 mm for 90Sr-90Y. At greater distances the measured values are larger than those calculated. For the concave 106Ru-106Rh applicator, there was poor agreement among available calculations and only those calculated by ACCEPT 3.0 agreed with measured values. In the past, attempts have been made to derive such dose distributions simply, by integrating the appropriate point-source dose function over the source. Here, we investigated the accuracy of this procedure for encapsulated sources, by comparing such results with values calculated by Monte Carlo. An attempt was made to allow for the effects of the silver source window but no corrections were made for scattering from the source backing. In these circumstances, at 6 mm depth, the difference in the results of the two calculations was 14%-18% for a planar 106Ru-l06Rh applicator and up to 30% for the concave applicator. It becomes worse at greater depths. These errors are probably caused mainly by differences between the spectrum of beta particles transmitted by the silver window and those transmitted by a thickness of water having the same attenuation properties. PMID- 11488570 TI - A dose-point-kernel model for a low energy gamma-emitting stent in a heterogeneous medium. AB - A computer dose model for a low energy gamma-emitting stent in a heterogeneous medium is described. The method is based on the Sievert model which is adapted to the dose-point-kernel (DPK) model to compute the dose distribution about filtered gamma sources (Sievert-DPK model). The new gamma stent model can take into account effects such as the metallic wire attenuation and the presence of dense calcified plaque in a stented artery. The Sievert-DPK model is tested against numerical simulations around cylindrical shell sources with dimensions comparable to those of a stent using a Monte Carlo transport code. For low energy gamma sources (Cs-131 and Pd-103), it is shown that the Sievert-DPK model is consistent with the Monte Carlo results to about 5%-10% for distances up to 5 mm from the cylindrical surface and 2.5 mm beyond the cylinder edges. These results indicate that the Sievert-DPK model may be useful to predict the dose in intravascular therapy applications for heterogeneous systems consisting of soft tissue, metal and dense plaque. PMID- 11488571 TI - Technical note: check software for use with trans-rectal ultrasound guided I-125 seed prostate implants. AB - In-house software has been developed to enable the dose distribution of a commercial treatment planning system to be verified prior to permanent trans rectal ultrasound guided I-125 seed implantation of the prostate. Printouts enabling the required loading pattern to be communicated to other staff groups are also generated. This software can be obtained via the authors and the AAPM software exchange (URL: http://aapm.org/medphys/resources/software/index.htm). PMID- 11488572 TI - An algorithm for automatic, computed-tomography-based source localization after prostate implant. AB - Permanent implant of the prostate using I-125 and Pd-103 seeds is a popular choice of treatment for early-stage prostate cancer in the United States. Evaluation of the quality of the implant is best based on the calculated dose distribution from postimplant computed tomography (CT) images. This task, however, has been time-consuming and inaccurate. We have developed an algorithm for automatic source localization from postimplant CT images. The only requirement of this algorithm is knowledge of the number of seeds present in the prostate, thus minimizing the need for human intervention. The algorithm processes volumetric CT data from the patient, and pixels of higher CT numbers are categorized into classes of definite and potential source pixels. A multithresholding technique is used to further determine the number of seeds and their precise locations in the CT volume data. A graphic user interface was developed to facilitate operator review of and intervention in the calculation and the results of the algorithm. This algorithm was tested on two phantoms containing nonradioactive seeds, one with 20 seeds in discrete locations and another with 100 seeds with small distances between seeds. The tests showed that the algorithm was able to identify the seed locations to within 1 mm of their physical locations for discrete seed locations. It was further able to separate seeds at close proximity to each other while maintaining an average seed localization error of less than 2 mm, with no operator intervention required. PMID- 11488573 TI - Dosimetry close to an 192Ir HDR source using N-vinylpyrrolidone based polymer gels and magnetic resonance imaging. AB - In this work, the utilization of polymer gel-MRI dosimetry for measurements at distances relevant to clinical brachytherapy and intravascular applications [i.e., in the mm range, where steep three-dimensional (3-D) dose gradients exist] is investigated using N-vinylpyrrolidone-based gels. Transverse axis radial dose distributions, dose distributions parallel to the source axis, and 2-D dose distributions around the commonly used microSelectron 192Ir HDR source are measured for single source dwell position irradiations. Experimental results are found in good agreement with verified Monte Carlo calculations, even for distances less than 3 mm from the source. The effect of various MRI parameters, such as slice thickness, slice mispositioning, and in-plane resolution, on the accuracy of the method is also investigated. Possible limitations of the method are discussed, and its' overall potential in brachytherapy dosimetry is evaluated. Experimental 2-D dose distributions for an intravascular application following the Paris irradiation protocol are compared to corresponding commercial treatment planning system calculations. Results suggest that polymer gel-MRI dosimetry is capable of experimentally verifying dose distributions in relevant clinical intravascular applications. PMID- 11488574 TI - A compact ridge filter for spread out Bragg peak production in pulsed proton clinical beams. AB - A number of designs have been proposed for ridge filters and range modulators used in proton therapy to modify the beam in order to spread out the Bragg peak. Despite the variety of solutions, no simple design capable of providing large fields and easy variation of the spread out Bragg peak (SOBP) length in a pulsed beam has been developed. We propose a compact ridge filter that can be used in a proton beam of any time structure. It allows the production of depth dose distributions that meet the requirements of therapy dose fields. PMID- 11488575 TI - Dosimetry of pulsed clinical proton beams by a small ionization chamber. AB - Response of a micro volume (0.01 ml) ionization chamber has been studied with pulsed proton beams which are used for clinical purposes and has been compared with those of some JARP ionization chambers (0.6 ml). All chambers used had been calibrated by standard 60Co beams at the Electrotechnical Laboratory (ETL) and exposure calibration factors, Nx, were obtained on advance. Two methods are used to compensate the general recombination which occurs during pulsed beam irradiations: theoretical correction by a Boag's formulation and a modified two voltage technique. An evaluation of absolute absorbed dose-to-water is performed on the basis of the protocol provided by ICRU report 59. The results imply that, to a first approximation, both chambers indicate the almost same result within 2% when unknown chamber-dependent parameters of the micro chamber are tentatively assumed to be identical to those of the JARP chamber for the calibration with 60Co beams. The about 1.5% discrepancy observed in the response of both chambers is not discussible due to presumably 1-2% uncertainty of the protocol of ICRU report 59 which does not include any chamber-dependent corrections for the perturbation effects in proton beams. PMID- 11488576 TI - High resolution gel-dosimetry by optical-CT and MR scanning. AB - The increased intricacy of Intensity-Modulated-Radiation-Therapy (IMRT) delivery has created the need for a high-resolution 3D-dosimetry (three-dimensional) system capable of measuring and verifying the complex delivery. Present clinical methods are inadequate being restricted to single points (e.g., ion-chambers) or to 2D planes (e.g., film), and are labor intensive. In this paper we show that gel-dosimetry in conjunction with optical-CT scanning can yield maps of dose that are of sufficient accuracy, resolution and precision to allow verification of complex radiosurgery deliveries, and by extension IMRT deliveries. The radiosurgery dose-distribution represents the most challenging case encountered in external beam therapy by virtue of the steep dose-gradients and high resolution of delivery. We characterize the stringent radiosurgery requirements by the RTAP (Resolution-Time-Accuracy-Precision) criteria defined as < or = 1 mm3 spatial resolution, < or = 1 hour imaging time, accurate to within 3%, and within -1% precision. The RTAP criteria is applied to an in-house laser-based optical-CT scanning system presented here, and evaluated using gel-flasks containing BANG3 gel. The same gel flasks were subsequently imaged using the MR imaging protocol recommended by the gel manufacturer, but modified to match as closely as possible the RTAP. The resulting dose-maps demonstrate the high precision (< 1.3% noise at high dose) achievable with optical CT scanning while preserving high spatial resolution (<1 mm3). Using the sequence above, the MR gel-dose maps were found to have poorer precision by a factor of 5, under the strict conditions of the RTAP. The optical CT gel-dosimetry system was further evaluated for the verification of a complex 3-isocenter radiosurgery delivery. In conclusion, this work demonstrates that gel-dosimetry and optical-CT scanning approach an important long-term goal of radiation dosimetry, as specified by the RTAP criteria, and have potential to impact the clinic by improving and facilitating clinical dose verification for the most complex external beam radiation treatments. PMID- 11488577 TI - The use of an inexpensive red acetate filter to improve the sensitivity of GAFChromic dosimetry. AB - The sensitivity of GAFChromic dosimetry using a conventional broad band light source densitometer has significantly been improved twofold using an inexpensive red acetate filter overlay during the densitometric measurements. This thin sheet of red acetate enhances the dosimetric analysis of radiochromic blue image distributions recorded on GAFChromic films. The combination provides higher sensitivity in the optical density measurements than the more expensive He-Ne laser-scanning densitometers. PMID- 11488578 TI - A survey of clinical factors and patient dose in mammography. AB - A survey was conducted to estimate the mean glandular dose (MGD) for women undergoing mammography and to report the distribution of doses, compressed breast thickness, glandular tissue content, and mammographic technique factors used. From 24,471 mammograms, of 6,006 women, clinical data were collected. The survey data included mammograms from seven modern units using a molybdenum (Mo) anode and either Mo or rhodium (Rh) filter. Exposure factors for each mammogram were recorded automatically onto a floppy disk on each unit. All mammography units were calibrated individually using breast tissue equivalent attenuation slabs of varying glandular content, so the breast glandular content could be estimated on the basis of exposure factors and compressed breast thickness. The MGD was estimated for each mammogram based on the normalized glandular dose and calculated entrance exposure in air. The survey found a median MGD of 2.6 mGy. The median breast glandular tissue content was 28% and the median compressed breast thickness was 5.1 cm. Also, patient attenuation data were converted to equivalent BR-12 and acrylic thickness to help determine appropriate phantom thicknesses required for mammography unit automatic exposure control performance assessment. PMID- 11488579 TI - Improvement of mammographic mass characterization using spiculation meausures and morphological features. AB - We are developing new computer vision techniques for characterization of breast masses on mammograms. We had previously developed a characterization method based on texture features. The goal of the present work was to improve our characterization method by making use of morphological features. Toward this goal, we have developed a fully automated, three-stage segmentation method that includes clustering, active contour, and spiculation detection stages. After segmentation, morphological features describing the shape of the mass were extracted. Texture features were also extracted from a band of pixels surrounding the mass. Stepwise feature selection and linear discriminant analysis were employed in the morphological, texture, and combined feature spaces for classifier design. The classification accuracy was evaluated using the area Az under the receiver operating characteristic curve. A data set containing 249 films from 102 patients was used. When the leave-one-case-out method was applied to partition the data set into trainers and testers, the average test Az for the task of classifying the mass on a single mammographic view was 0.83 +/- 0.02, 0.84 +/- 0.02, and 0.87 +/- 0.02 in the morphological, texture, and combined feature spaces, respectively. The improvement obtained by supplementing texture features with morphological features in classification was statistically significant (p = 0.04). For classifying a mass as malignant or benign, we combined the leave-one-case-out discriminant scores from different views of a mass to obtain a summary score. In this task, the test Az value using the combined feature space was 0.91 +/- 0.02. Our results indicate that combining texture features with morphological features extracted from automatically segmented mass boundaries will be an effective approach for computer-aided characterization of mammographic masses. PMID- 11488580 TI - Half-scan cone-beam CT fluoroscopy with multiple x-ray sources. AB - To develop volumetric micro-CT fluoroscopy for small animal imaging, we have proposed a cone-beam system with multiple x-ray sources. In this paper, we extend Parker's single-source half-scan weighting scheme to the case of an odd number of x-ray sources that are equiangularly distributed, and apply it for half-scan Feldkamp-type reconstruction in this unique geometry. In the numerical simulation with the Shepp-Logan phantom, representative images indicate that the proposed half-scan Feldkamp-type algorithm produces temporal resolution significantly superior to that with a single x-ray source cone-beam system. PMID- 11488581 TI - Alignment of a volumetric tomography system. AB - A test-bed system has been developed for imaging phantoms with tomosynthesis and volumetric computed tomography. This system incorporates an amorphous silicon flat panel detector on a movable gantry and a computer-controlled rotational positioning stage. In this paper, an analysis of the sensitivity of reconstructed images to geometrical misalignment is presented. Application of this method to circular digital tomosynthesis is examined, with spatial resolution in the focal plane as the criterion for evaluating the effect of misalignment. A software based method is presented for correcting data for imperfect system alignment prior to image reconstruction. Experimental results yield reconstructed images with spatial resolution approaching the theoretical limit based on detector pixel size and accounting for data interpolation. PMID- 11488582 TI - Digital subtraction peripheral angiography using image stacking: initial clinical results. AB - Using clinically acquired x-ray angiography image sequences, we compared three algorithms for creating a single diagnostic quality image that combined input images containing flowing contrast agent. These image-stacking algorithms were: maximum opacity with the minimum gray-scale value across time recorded at each spatial location, (REC) recursive temporal filtering followed by a maximum opacity operation, and (AMF) an approximate matched filter consisting of a convolution with a kernel approximating the matched filter followed by a maximum opacity operation. Eighteen clinical exams of the peripheral arteries of the legs were evaluated. AMF gave 2.7 times greater contrast to noise ratio than the single best subtraction image and 1.3 times improvement over REC, the second best stacking algorithm. This is consistent with previous simulations showing that AMF performs nearly equal to the optimal result from matched filtering without the well-known limitations. For example, unlike matched filtering, AMF filter coefficients were obtained automatically using an image-processing algorithm. AMF effectively brought out small collateral arteries, otherwise difficult to see, without degrading artery sharpness or stenosis grading. Comparing results using reduced and full contrast agent volumes demonstrated that contrast agent load could be reduced to one-third of the conventional amount with AMF processing. By simulating reduced x-ray exposures on clinical exams, we determined that x-ray exposure could be reduced by 80% with AMF processing. We conclude that AMF is a promising, potential technique for reducing contrast agent load and for improving vessel visibility, both very important characteristics for vascular imaging. PMID- 11488583 TI - Chord distributions across 3D digital images of a human thoracic vertebra. AB - Radiation dose estimates to the trabecular region of the skeleton are of primary importance due to recent advancements in nuclear medicine. Establishing methods for accurately calculating dose in these regions is difficult due to the complex microstructure of this anatomic site and the typical ranges of beta-particles in both bone and marrow tissues. At the present time, models of skeletal dosimetry used in clinical medicine rely upon measured distributions of straight-line path lengths (chord lengths) through bone and marrow regions. This work develops a new three-dimensional, digital method for acquiring these distributions within voxelized images. In addition, the study details the characteristics of measuring chord distributions within digital images and provides a methodology for avoiding undesirable pixel or voxel effects. The improved methodology has been applied to a digital image (acquired via NMR microscopy) of the trabecular region of a human thoracic vertebra. The resulting chord-length distributions across both bone trabeculae and bone marrow cavities were found to be in general agreement with those measured in other studies utilizing different methods. In addition, this study identified that bone and marrow space chord-length distributions are not statistically independent, a condition implicitly assumed within all current skeletal dosimetry models of electron transport. The study concludes that the use of NMR microscopy combined with the digital measurement techniques should be used to further expand the existing Reference Man database of trabecular chord distributions to permit the development of skeletal dosimetry models which are more age and gender specific. PMID- 11488584 TI - Beta-particle dosimetry of the trabecular skeleton using Monte Carlo transport within 3D digital images. AB - Presently, skeletal dosimetry models utilized in clinical medicine simulate electron path lengths through skeletal regions based upon distributions of linear chords measured across bone trabeculae and marrow cavities. In this work, a human thoracic vertebra has been imaged via nuclear magnetic resonance (NMR) spectroscopy yielding a three-dimensional voxelized representation of this skeletal site. The image was then coupled to the radiation transport code EGS4 allowing for 3D tracing of electron paths within its true 3D structure. The macroscopic boundaries of the trabecular regions, as well as the cortex of cortical bone surrounding the bone site, were explicitly considered in the voxelized transport model. For the case of a thoracic vertebra, energy escape to the cortical bone became significant at source energies exceeding approximately 2 MeV. Chord-length distributions were acquired from the same NMR image, and subsequently used as input for a chord-based dosimetry model. Differences were observed in the absorbed fractions given by the chord-based model and the voxel transport model, suggesting that some of the input chord distributions for the chord-based models may not be accurate. Finally, this work shows that skeletal mass estimates can be made from the same NMR image in which particle transport is performed. This feature allows one to determine a skeletal S-value using absorbed fraction and mass data taken from the same anatomical tissue sample. The techniques developed in this work may be applied to a variety of skeletal sites, thus allowing for the development of skeletal dosimetry models at all skeletal sites for both males and females and as a function of subject age. PMID- 11488585 TI - Signal processing in scanning thermoacoustic tomography in biological tissues. AB - Microwave-induced thermoacoustic tomography was explored to image biological tissues. Short microwave pulses irradiated tissues to generate acoustic waves by thermoelastic expansion. The microwave-induced thermoacoustic waves were detected with a focused ultrasonic transducer to obtain two-dimensional tomographic images of biological tissues. The dependence of the axial and the lateral resolutions on the spectra of the signals was studied. A reshaping filter was applied to the temporal piezoelectric signals from the transducer to increase the weight of the high-frequency components, which improved the lateral resolution, and to broaden the spectrum of the signal, which enhanced the axial resolution. A numerical simulation validated our signal-processing approach. PMID- 11488586 TI - Evaluation of multielement catheter-cooled interstitial ultrasound applicators for high-temperature thermal therapy. AB - Catheter-cooled (CC) interstitial ultrasound applicators were evaluated for their use in high-temperature coagulative thermal therapy of tissue. Studies in ex vivo beef muscle were conducted to determine the influences of applied electrical power levels (5-20 W per element), catheter flow rate (20-60 ml min(-1)), circulating water temperature (7-40 degrees C), and frequency (7-9 MHz) on temperature distribution and thermal lesion geometry. The feasibility of using multiple interstitial applicators to thermally coagulate a predetermined volume of tissue was also investigated. Results of these studies revealed that the directional shape of the thermal lesions is maintained with increasing time and power. Radial depths of the thermal lesions ranged from 10.7 +/- 0.7 mm after heating for 4 min with an applied power level of 5 W, to 16.2 +/- 1.4 mm with 20 W. The axial length of the thermal lesions is controlled tightly by the number of active transducers. A catheter flow rate of 20 to 40 ml min(-1) (52.2 +/- 5.5 kPa at 40 ml min(-1)) with 22 degrees C water was determined to provide sufficient cooling of the transducers for power levels used in this study. In vivo temperatures measured in the center of a 3-cm-diam peripheral implant of four applicators in pig thigh muscle reached 89.3 degrees C after 4 min of heating, with boundaries of coagulation clearly defined by applicator position and directivity. Conformability of heating in a clinically relevant model was demonstrated by inserting two directional CC applicators with a 2 cm separation within an in vivo canine prostate, and generating a thermal lesion measuring 3.8 cm x 2.2 cm in cross section while directing energy away from, and protecting the rectum. Maximum measured temperatures at midgland exceeded 90 degrees C within 20 min of heating. The results of this study demonstrate the utility of single or multiple CC applicators for conformal thermal coagulation and high temperature thermal therapy, with potential for clinical applications in sites such as prostate, liver, breast, or uterus. PMID- 11488587 TI - Medical litigation in Ireland. PMID- 11488588 TI - Filariasis control: ethics, economics, and good science. PMID- 11488589 TI - Skin substitutes--benefits and costs. PMID- 11488590 TI - Specific electrochemical nitration of horse heart myoglobin. AB - Earlier findings on electronitration of hen egg-white lysozyme demonstrated a product which was mononitrated at Tyr23, by ion-exchange chromatography, absorbance at 430 nm, dithionite reduction, and Edman sequencing of a nitrated proteolytic peptide. However, the whole protein was not sequenced; therefore, although the enzyme remained active upon nitration, reaction at other residues could not be completely eliminated. This study has now been extended to the redox protein myoglobin. We demonstrate the novel electronitration (electrooxidation in the presence of nitrite) of a specific tyrosine residue in horse heart myoglobin and also in apomyoglobin. Production of the yellow chromophore, 3-nitrotyrosine (3-NT), was apparent in apomyoglobin from A430 but was masked in holomyoglobin by the Soret band. In both cases, the presence of 3-NT in the electronitrated samples was further indicated by the binding of antibody to 3-NT in Western blots. High-resolution electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry revealed a reaction product at [M + 45] (consistent with substitution of NO2 for H), indicating that the nitration reaction is the only reaction occurring which gives rise to a change in mass in the electrooxidation. Fragmentation mass spectrometry identified the nitration site as Tyr103, with no nitration at Tyr146. The procedure may be useful in preparing model nitrated proteins for the study of disease mechanisms. PMID- 11488591 TI - Potential involvement of several nitroreductases in metronidazole resistance in Helicobacter pylori. AB - Susceptibility of Helicobacter pylori to the antibiotic metronidazole has been attributed to the activity of an oxygen-insensitive NADPH-dependent nitroreductase (RdxA), with resistance to this antimicrobial arising from null mutations in rdxA. To obtain a better understanding of the factors involved in resistance, nitroreductase and metronidazole reduction activities were investigated in matched pairs of clinical and laboratory-derived sensitive and resistant H. pylori strains. Significant differences in enzyme activities were observed between sensitive and resistant strains, suggesting that metronidazole susceptibility in H. pylori was associated with more than one enzyme activity. To establish the mutations occurring in rdxA, the genes from seventeen bacterial strains, including matched pairs were sequenced. To assess whether metronidazole was responsible for inducing random mutations in this gene, the complete nucleotide sequence of gene hp0630, encoding an NAD(P)H-quinone reductase which also has NADPH-dependent nitroreductase activity, was determined in the same strains. All resistant strains showed nonsense, missense, or frameshift mutations randomly throughout rdxA. In contrast, no mutations were observed in hp0630. The results confirmed the presence of rdxA null mutations in resistant strains and suggested that other factors involved in the metabolism of metronidazole contributed to the resistant phenotype. PMID- 11488592 TI - Oxidative stress and heat shock stimulate RGS2 expression in 1321N1 astrocytoma cells. AB - RGS2, a regulators of G-protein signaling family member, regulates G-protein signaling and is itself controlled in part by regulated expression. We tested if cell stress regulates RGS2 expression in human astrocytoma 1321N1 cells. Treatment with H2O2 increased RGS2 mRNA levels time- and concentration dependently, with 200 microM H2O2 causing an approximately eightfold increase after 2 h. Peroxynitrite and heat shock also increased RGS2 mRNA levels. H2O2 induced RGS2 expression was negatively regulated by phosphoinositide-3-kinase and extracellular signal-regulated kinases. H2O2 also concentration-dependently increased RGS2 protein levels, and the RGS2 appeared to be predominantly in the nucleus. These results demonstrate that RGS2 expression is up-regulated by cell stress. PMID- 11488593 TI - Transfection of mGSTA4 in HL-60 cells protects against 4-hydroxynonenal-induced apoptosis by inhibiting JNK-mediated signaling. AB - The mammalian alpha-class glutathione S-transferase (GST) isozymes mGSTA4-4, rGSTA4-4, and hGSTA4-4 are known to utilize 4-hydroxynonenal (4HNE) as a preferred substrate. During the present studies, we have examined the effect of transfecting human myeloid HL-60 cells with mGSTA4, on 4-HNE-induced apoptosis and the associated signaling mechanisms. Results of these studies show that treatment of the wild-type or vector-only-transfected HL-60 cells with 20 microM 4-HNE caused apoptosis within 2 h. The cells transfected with mGSTA4 did not undergo apoptosis under these conditions even after 4 h. In the wild-type and vector-transfected cells, apoptosis was preceded by JNK activation and c-Jun phosphorylation within 30 min, and an increase in AP-1 binding within 2 h of treatment with 20 microM 4-HNE. In mGSTA4-transfected cells, JNK activation and c Jun phosphorylation were observed after 1 h, and increased AP-1 binding was observed after 8 h under these conditions. In the control cells, 20 microM 4-HNE caused caspase 3 activation and poly(ADP-ribose) polymerase cleavage within 2 h, while in mGSTA4-transfected cells, a lesser degree of these effects was observed even after 8 h. Transfection with mGSTA4 also provided protection to the cells from 4-HNE and doxorubicin cytotoxicity (1.6- and 2.6-fold, respectively). These results show that 4-HNE mediates apoptosis through its effects on JNK and caspase 3, and that 4-HNE metabolizing GST isozyme(s) may be important in the regulation of this pathway of oxidative-stress-induced apoptosis. PMID- 11488594 TI - Biophysical characterization of a soluble CD40 ligand (CD154) coiled-coil trimer: evidence of a reversible acid-denatured molten globule. AB - The CD40 ligand molecule is unique, consisting of a receptor-binding domain anchored by an isoleucine zipper moiety. Exact determination of the multimeric state and its tendency to form molten globules has not been elucidated. Corroborating evidence of a trimerized molecule in aqueous solution was obtained from size-exclusion chromatography, laser light scattering, and analytical ultracentrifugation. A reversible acid-denatured molten globule state was observed from circular dichroism and fluorescence spectroscopy data. The molten globule state was characterized by a loss of tertiary structure with associated retention of secondary structure near pH 3. Once returned to pH 7, the acid denatured state refolded over the course of 7 days resulting in approximately 90% recovery of the native structure. The molten globule state was characterized by a broadening of structural features in the second-derivative spectra of Fourier transform infrared spectroscopy. A component band at 1650 cm(-1) was shown to be alpha-helix and originate from amide carbonyl vibrations of the isoleucine zipper. Differential scanning calorimetry measurements characterized the pH sensitive molten globule state at pH 3.3 as one lacking a well-defined unfolding transition with an accompanying baseline shift at 58 degrees C (a consequence of increased heat capacity). The tendency to form molten globules during acid denaturation stress permits an opportunity to study the process of partial protein unfolding with implications concerning stability. Although reversible molten globules can be formed, it is important to recognize the unusual nature since the molten globule state is formed exclusively within the beta-sheet receptor-binding region. PMID- 11488595 TI - Purification of diacylglycerol kinase from Microsporum gypseum and its phosphorylation by the catalytic subunit of protein kinase A. AB - Diacylglycerol (DG) kinase (EC 2.7.1.107) was purified to homogeneity from the soluble extract of Microsporum gypseum, a dermatophyte. Purified enzyme showed a final specific activity of 2172 pmol/min/mg protein and its apparent molecular weight on SDS-PAGE was found to be 93 kDa. The activity of purified enzyme was inhibited in a dose-dependent manner in the presence of DG-kinase inhibitor (D5919, Sigma). DG-kinase activity was found to be stimulated in the presence of phosphatidylcholine, phosphatidylethanolamine, and cardiolipin while the activity was alleviated in the presence of phosphatidic acid and arachidonic acid. Kinase activity was partially inhibited when assayed after prior treatment with alkaline phosphatase. Treatment of DG-kinase with the catalytic subunit of protein kinase A (PKA)-stimulated DG-kinase activity in a dose-dependent manner. Incubation of DG-kinase with the catalytic subunit of PKA led to the phosphorylation of DG kinase as revealed by autoradiography. The phosphorylated band disappeared completely in the presence of specific PKA inhibitor. Increased activity of DG kinase on incubation with the catalytic subunit of PKA was possibly due to the phosphorylation of the former by the latter. Whether this in vitro phosphorylation and activation of DG-kinase occurs under physiological conditions remains to be elucidated. PMID- 11488596 TI - Disruption of histidine catabolism in NEUT2 mice. AB - Homozygous NEUT2 mice lack cytosolic 10-formyltetrahydrofolate dehydrogenase (FDH; Champion et al. (1994) Proc. Natl. Acad. Sci. USA 91, 11,338-11,342) and as a consequence should be unable to oxidize carbon 2 of l-histidine to CO2 via 10 formyltetrahydrofolate in liver cytosol. There was essentially no oxidation of l [2-14C]histidine to 14CO2 in homozygous NEUT2 mice, but 52% of the [2-14C]l histidine dose was recovered in the urine within 24 h. Analysis of urine samples for [14C]formiminoglutamate, the expected excretion product, was negative; however, [14C]urocanic acid was detected. Investigation of histidine catabolism via the folate-dependent deamination pathway revealed no detectable urocanase activity in homozygous NEUT2 mice, while heterozygous NEUT2 mice had 50% urocanase activity compared to normal mice. Histidase and formiminotransferase cyclodeaminase, also on the histidine deamination pathway, had similar specific activities in normal and NEUT2 mice. Histidine-pyruvate aminotransferase, the first enzyme of the alternate histidine transamination catabolic pathway did not appear to be affected by the loss of urocanase. Based on the excretion of urocanic acid it is estimated that NEUT2 mice catabolize approximately 40 micromol/day via the deamination pathway. The loss of urocanase activity in homozygous NEUT2 mice may allow these mice to survive the disruption in folate metabolism by sparing the liver cytosolic tetrahydrofolate pool. PMID- 11488597 TI - Absolute kinetic characterization of 17-beta-estradiol as a radical-scavenging, antioxidant synergist. AB - We directly measured the absolute reactivity of 17-beta-estradiol (E2) and several phenolic model compounds for E2 toward t-butoxy radical (t-BuO*) by nanosecond time-resolved optical spectroscopy. Compared to other phenols, E2 is a moderate, but not strong deactivator of oxyradicals. The absolute bimolecular rate constant for H-atom transfer from E2 to t-BuO* is 1.3 +/- 0.3 x 10(9) M(-1) x s(-1) (23 degrees C, benzene). We estimate the O-H bond strength of 17-beta estradiol to be approximately 85 +/- 2 kcal/mol and calculate the reaction rate constant of E2 toward peroxy radical to be 10(5) M(-1) x s(-1) at 37 degrees C. The conjugate phenoxy radical of 17-beta-estradiol, E2O*, is unusually reactive toward alpha-tocopherol and ascorbate by H-atom transfer in homogeneous solution (10(8)-10(9) M(-1) x s(-1)). Our findings suggest that E2 functions in vivo as a highly localized, synergistic biological antioxidant. This may partly explain the clinical effectiveness of ovarian steroids in delaying the manifestations of Alzheimer's Disease as well as in protecting against cardiovascular pathologies. In the absence of complementary antioxidant synergists, E2O* is expected to be a pro-oxidant. PMID- 11488598 TI - Cationic liposomes induce macrophage apoptosis through mitochondrial pathway. AB - To clarify the mechanism of apoptosis of the macrophage-like cell line RAW264.7 induced by cationic liposomes, we focused on the mitochondria and investigated the changes in mitochondrial membrane potential and the release of cytochrome c following treatment of cationic liposomes composed of stearylamine (SA liposomes). SA-liposomes induced mitochondrial membrane depolarization and also the release of cytochrome c from mitochondria. Caspase-3 was also activated by SA liposome treatment. Pretreatment of cells with N-acetylcysteine, a scavenger of reactive oxygen species (ROS), conferred resistance to the induction of the membrane depolarization, cytochrome c release, and caspase-3 activation by SA liposomes. These results indicated that SA-liposomes caused the apoptosis in RAW264.7 cells through the mitochondrial pathway, and ROS generation was required for this phenomenon. PMID- 11488599 TI - Surface NADH oxidase of HeLa cells lacks intrinsic membrane binding motifs. AB - Disulfide-thiol interchange proteins with hydroquinone (NADH) oxidase activities (designated NOX for plasma membrane-associated NADH oxidases) occur as extrinsic membrane proteins associated with the plasma membrane at the outer cell surface. The cancer-associated NOX protein, designated tNOX, has been cloned. The 34-kDa plasma membrane-associated form of the protein contains no strongly hydrophobic regions and is not transmembrane. No myristoylation or phosphatidylinositol anchor motifs were discovered. Evidence for lack of involvement of a glycosylphosphatidylinositol-linkage was derived from the inability of treatment with a phosphatidylinositol-specific phospholipase C or with nitrous acid at low pH to release the NOX protein from the surface of HeLa cells or from plasma membranes isolated from HeLa cells. Binding of NOX protein to the plasma membrane via amino acid side chain modification or by attachment of fatty acids also is unlikely based on use of specific fatty acid antisera to protein bound fatty acids and as a result of binding to the cancer cell surface of a truncated form of recombinant tNOX. Incubation of cells or plasma membranes with 0.1 M sodium acetate, pH 5, at 37 degrees C for 1 h, was sufficient to release tNOX from the HeLa cell surface. Release was unaffected by protease inhibitors or divalent ions and was not accelerated by addition of cathepsin D. The findings suggest dissociable receptor binding as a possible basis for their plasma membrane association. PMID- 11488600 TI - Role of Ca2+-independent phospholipase A2 on arachidonic acid release induced by reactive oxygen species. AB - Previous studies have shown that reactive oxygen species (ROS) enhance arachidonic acid (AA) release and the subsequent AA metabolism in macrophages. The purpose of this study was determined the implication of phospholipases A2 (PLA2s) in these events. Our results show that oxidative stress induced by exogenous adding of hydrogen peroxide or superoxide anion in macrophage RAW 264.7 and mouse peritoneal macrophage cultures caused a marked enhancement of calcium independent PLA2 (iPLA2) activity,whereas the increment of secreted PLA2 (sPLA2) and calcium-dependent cytosolic PLA2 (cPLA2) activities were slight. This increase of iPLA2 activity by ROS was rapid and dose-dependent. ROS also induced a significant [3H] arachidonic acid (AA) release. The iPLA2 selective inhibitor, bromoenol lactone, almost completely suppressed the mobilization of [3H]AA induced by ROS whereas antisense oligonucleotide against cPLA2 did not have any appreciable effect. Thus, our data show that iPLA2 activity is involved in the mechanism by which ROS increases the availability of free AA in macrophages RAW 264.7. Moreover, the protein kinase C (PKC) inhibitor, calphostin C, and calcium chelators had no effect on the [3H]AA release induced by ROS, suggesting this is a regulatory role of iPLA2. PMID- 11488601 TI - Cloning and characterization of Ginkgo biloba levopimaradiene synthase which catalyzes the first committed step in ginkgolide biosynthesis. AB - Levopimaradiene synthase, which catalyzes the initial cyclization step in ginkgolide biosynthesis, was cloned and functionally characterized. A Ginkgo biloba cDNA library was prepared from seedling roots and a probe was amplified using primers corresponding to conserved gymnosperm terpene synthase sequences. Colony hybridization and rapid amplification of cDNA ends yielded a full-length clone encoding a predicted protein (873 amino acids, 100,289 Da) similar to known gymnosperm diterpene synthases. The sequence includes a putative N-terminal plastid transit peptide and three aspartate-rich regions. The full-length protein expressed in Escherichia coli cyclized geranylgeranyl diphosphate to levopimaradiene, which was identical to a synthetic standard by GC/MS analysis. Removing 60 or 79 N-terminal residues increased levopimaradiene production, but a 128-residue N-terminal deletion lacked detectable activity. This is the first cloned ginkgolide biosynthetic gene and the first in vitro observation of an isolated ginkgolide biosynthetic enzyme. PMID- 11488602 TI - Accessibility and activity of the promoter for a dioxin-inducible ecto-ATPase gene. AB - We have analyzed the core promoter for a dioxin-inducible ecto-ATPase gene in mouse hepatoma cells. The transcriptional initiation site maps to a region that contains neither a TATA sequence nor a consensus initiator sequence nor a downstream promoter element. The core promoter has constitutive activity that does not require either the aromatic hydrocarbon receptor or its heterodimerization partner Arnt. Two GC-rich regions contribute approximately equally to the constitutive activity. Proteins constitutively occupy the GC-rich regions in chromatin. The promoter assumes a non-nucleosomal configuration in its native chromosomal setting in both uninduced and dioxin-induced cells. Our findings imply that the GC-rich regions together with their cognate binding proteins carry out core promoter functions for the ecto-ATPase gene. The promoter is constitutively accessible in situ, and chromatin structure is not a limiting factor for dioxin-inducible ecto-ATPase transcription in intact cells. PMID- 11488603 TI - Characterization of a novel laccase produced by the wood-rotting fungus Phellinus ribis. AB - The white-rot fungus Phellinus ribis produced a single form of laccase, which was purified to apparent electrophoretic homogeneity from cultures induced with 2,5 xylidine. This protein was a dimer, consisting of two subunits of 76 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Carbohydrate analysis revealed that the enzyme contained about 28% carbohydrate content. The laccase appeared to be different from other known laccases by the UV visible absorption spectrum analysis. One enzyme molecule contained one copper, one manganese, and two zinc atoms. The laccase showed optimal activity at pH 4.0 6.0, 5.0, and 6.0 with 2,6-dimethoxyphenol, ABTS [2,2'-azino-bis-(3 ethylbenzothiazoline-6-sulfonic acid)], and syringaldazine, respectively. The enzyme preferably oxidized dimethoxyphenol and aromatic amine compounds. The stability of the laccase was low at acidic pH, whereas it showed high stability at neutral pH and mild temperature. The N-terminal amino acid sequence revealed a very low homology with other microbial laccases. With some substrates, the addition of manganese and H2O2 resulted in a remarkable increase in the oxidation rate. Without an appropriate phenolic substrate, the enzyme could not oxidize Mn(II) in the presence of H2O2 or pyrophosphate. PMID- 11488604 TI - In vitro assembly of feline immunodeficiency virus capsid protein: biological role of conserved cysteines. AB - Core assembly, a key step in the retroviral life cycle, is poorly understood. Previous studies have shown that the entire gag region is needed to form the assembled particles. In this report, we have shown that the assembly process is driven by recombinant capsid protein (p26) of feline immunodeficiency virus itself. Proteins are expressed in a bacterial system and soluble forms of wild type and modified proteins are purified from bacterial extracts and are examined on gel-filtration chromatography fitted to an HPLC system. It has also been shown that changing residue Cys190 (one of the two conserved cysteines of feline immunodeficiency virus which are also conserved for all the immunodeficiency viruses including HIV) to serine by site-directed mutagenesis disrupts the assembly process. In addition, this modification causes considerable thermal instability of the protein while substitutions at nonconserved cysteines do not significantly affect the thermal stability and assembly of the protein. These findings indicate that conserved cysteine residues play a vital role in the capsid protein assembly and, therefore, are critical for virus infectivity. PMID- 11488605 TI - Catalase-like oxygen production by horseradish peroxidase must predominantly be an enzyme-catalyzed reaction. AB - When hydrogen peroxide (H2O2) was provided as the only substrate for horseradish peroxidase C (HRP-C) the catalase-like emission of oxygen gas was observed. The reaction was favored at neutral compared to acidic pH. Addition of the superoxide radical scavengers tetranitromethane (TNM) or superoxide dismutase (SOD) increased activity. TNM's effect was concentration dependent but SOD's was not, indicating that only some of the superoxide generated was released into solution. Manganous ions (Mn2+) react with superoxide radicals to regenerate H2O2 but not oxygen; when added to the reaction medium oxygen production was reduced but not abolished. The effect was essentially concentration independent, suggesting that most oxygen was produced enzymatically and not by chemical disproportionation of superoxide. The catalase-like activities of some site-directed mutants of HRP-C suggest that active site residues histidine 42 and arginine 38 are influential in determining this activity. A clear correlation also existed between catalase activity and the enzymes' resistance to inactivation by H2O2. Computer simulation of a reaction scheme that included catalase-like activity agreed well with experimental data. PMID- 11488606 TI - Molecular and catalytic properties of Arabidopsis thaliana adenylyl sulfate (APS) kinase. AB - A cDNA clone (Atakn1) from Arabidopsis thaliana encoding APS-kinase (EC 2.7.1.25) was investigated for structural and catalytic properties of the gene product. Recombinant his10-AtAkn1 formed PAPS at a Vmax of 7.35 U x mg(-1). The Km for APS was 0.14 microM and for ATP 147 microM. APS caused a severe substrate inhibition (K(i) 4.5 microM). The type of inhibition is uncompetitive with respect to MgATP. High ionic strength and reducing thiols stabilized the enzyme activity. Plant APS kinase is regulated in vitro by the redox charge with thioredoxin as essential activator. Mutagenesis of a serine in S182C and S182F presumed to be involved in the transfer of the phosphoryl group had no effect upon catalytic activity. Using a yeast two-hybrid system with AtAkn1 as bait, an interacting clone was detected from a cDNA library of A. thaliana cv. Columbia that codes for an APS-kinase iso form (Atakn2). Complementation of APS-kinase-deficient Saccharomyces cerevisiae met14 showed that AtAkn2 is functionally active as APS-kinase. It was immunologically related to AtAkn1 and presumably represents a plastidal iso-form of the plant APS-kinase gene family. PMID- 11488607 TI - Vanadate-induced cell growth regulation and the role of reactive oxygen species. AB - While vanadium compounds are known as potent toxicants as well as carcinogens, the mechanisms of their toxic and carcinogenic actions remain to be investigated. It is believed that an improper cell growth regulation leads to cancer development. The present study examines the effects of vanadate on cell cycle control and involvement of reactive oxygen species (ROS) in these vanadate mediated responses in a human lung epithelial cell line, A549. Under vanadate stimulation, A549 cells generated hydroxyl radical (*OH), as determined by electron spin resonance (ESR), and hydrogen peroxide (H2O2) and superoxide anion (O2*-), as detected by flow cytometry using specific dyes. The mechanism of ROS generation involved the reduction of molecular oxygen to O2*- by both a flavoenzyme-containing NADPH complex and the mitochondria electron transport chain. The O2*- in turn generated H2O2, which reacted with vanadium(IV) to generate *OH radical through a Fenton-type reaction (V(IV) + H2O2 --> V(V) +*OH + OH-). The ROS generated by vanadate induced G2/M phase arrest in a time- and dose dependent manner as determined by measuring DNA content. Vanadate also increased p21 and Chk1 levels and reduced Cdc25C expression, leading to phosphorylation of Cdc2 and a slight increase in cyclin B1 expression as analyzed by Western blot. Catalase, a specific antioxidant for H2O2, decreased vanadate-induced expression of p21 and Chk1, reduced phosphorylation of Cdc2Tyr15, and decreased cyclin B1 levels. Superoxide dismutase, a scavenger of O2*-, or sodium formate, an inhibitor of *OH, had no significant effects. The results obtained from the present study demonstrate that among ROS, H2O2 is the species responsible for vanadate-induced G2/M phase arrest. Several regulatory pathways are involved: (1) activation of p21, (2) an increase of Chk1 expression and inhibition of Cdc25C, which results in phosphorylation of Cdc2 and possible inactivation of cyclin B1/Cdc2 complex. PMID- 11488608 TI - Rat liver mitochondrial contact sites and carnitine palmitoyltransferase-I. AB - In hepatic mitochondria, the outer membrane enzyme, carnitine palmitoyltransferase-I (CPT-I), appears to colocalize with contact sites. We have prepared contact sites that are essentially devoid of noncontact site membranes. The contact site fraction has a high specific activity for CPT-I and contains a protein at 88 kDa that is recognized by antibodies directed at two different peptide epitopes on CPT-I. Similarly long-chain acyl-CoA synthetase (LCAS) specific activity is high in this fraction; a protein at 79 kDa is recognized by an antibody against LCAS. Although activity of carnitine palmitoyltransferase-II (CPT-II) is present, it is not enriched in the contact site fraction, and a protein of 68 kDa weakly reacted with anti-CPT-II antibody. Likewise, carnitine acylcarnitine translocase (CACT) protein is present, but at a somewhat reduced level. Using an analytical continuous sucrose gradient, we demonstrate that the activities of CPT-I and LCAS and their associated immunoreactive proteins are present in a constant amount throughout the contact site subfractions. The enzymatic activity of CPT-II and its associated immunoreactive protein, as well as immunoreactive CACT, is absent in the lighter density gradient subfractions and is present in the higher density subfractions only in trace amounts. This heterogeneity of the contact site fraction is due to unvarying amounts of outer membrane and increasing amounts of attached inner membrane with increasing density of the subfractions. PMID- 11488609 TI - Requirements of Cyc2p and the porin, Por1p, for ionic stability and mitochondrial integrity in Saccharomyces cerevisiae. AB - It was previously demonstrated that Cyc2p from Saccharomyces cerevisiae is a mitochondrial protein; that the cyc2-Delta2 deletion lacking the entire gene causes a diminution to only approximately 20% of the normal levels of cytochrome c due to a partial deficiency in mitochondrial import of apo-cytochrome c; that the deletion causes a defective mitochondrial function, as revealed by diminished growth on media containing nonfermentable carbon sources; and that this defect is exacerbated in hyper-ionic KCl media and at higher incubation temperatures, but is suppressed on media containing sorbitol, a nonionic compound. We report that por1-Delta strains lacking the mitochondrial porin, Por1p, but not por2-Delta strains lacking the related porin, share some phenotypes similar to the cyc2 Delta2 strain, including hypersensitivity to KCl in glycerol medium. Moreover, spontaneous swelling in the presence of ATP was detected in mitochondria from the cyc2-Delta2 strain, while swelling could be detected in mitochondria from the other strains only after the addition of KCl. Thus, highly unspecific membrane permeation may be triggered by ATP in the cyc2-Delta2 strain. We suggest that Por1p and Cyc2p, in addition to their own unique functions, serve to maintain the osmotic stability of mitochondria, but by different mechanisms. PMID- 11488610 TI - Cofactor and tryptophan accessibility and unfolding of brain glutamate decarboxylase. AB - Cofactor and tryptophan accessibility of the 65-kDa form of rat brain glutamate decarboxylase (GAD) was investigated by fluorescence quenching measurements using acrylamide, I-, and Cs+ as the quenchers. Trp residues were partially exposed to solvent. I- was less able and Cs+ was more able to quench the fluorescence of Trp residues in the holoenzyme of GAD (holoGAD) than the apoenzyme (apoGAD). The fraction of exposed Trp residues were in the range of 30-49%. In contrast, pyridoxal-P bound to the active site of GAD was exposed to solvent. I- was more able and Cs+ was less able to quench the fluorescence of pyridoxal-P in holoGAD. The cofactor was present in a positively charged microenvironment, making it accessible for interactions with anions. A difference in the exposure of Trp residues and pyridoxal-P to these charged quenchers suggested that the exposed Trp residues were essentially located outside of the active site. Changes in the accessibility of Trp residues upon pyridoxal-P binding strongly supported a significant conformational change in GAD. Fluorescence intensity measurements were also carried out to investigate the unfolding of GAD using guanidine hydrochloride (GdnHCl) as the denaturant. At 0.8-1.5 M GdnHCl, an intermediate step was observed during the unfolding of GAD from the native to the denatured state, and was not found during the refolding of GAD from the denatured to native state, indicating that this intermediate step was not a reversible process. However, at >1.5 M GdnHCl for holoGAD and >2.0 M GdnHCl for apoGAD, the transition leading to the denatured state was reversible. It was suggested that the intermediate step involved the dissociation of native dimer of GAD into monomers and the change in the secondary structure of the protein. Circular dichroism revealed a decrease in the alpha-helix content of GAD from 36 to 28%. The unfolding pattern suggested that GAD may consist of at least two unfolding domains. Unfolding of the lower GdnHCl-resisting domain occurred at a similar concentration of denaturant for apoGAD and holoGAD, while unfolding of the higher GdnHCl-resisting domain occurred at a higher concentration of GdnHCl for apoGAD than holoGAD. PMID- 11488611 TI - Thioester enolate stabilization in the acyl-CoA dehydrogenases: the effect of 5 deaza-flavin substitution. AB - The redox-inactive thioester analog 3-thia-octanoyl-CoA blocks transfer of a hydride equivalent to the flavin prosthetic group of the medium-chain acyl-CoA dehydrogenase with the accumulation of a stable enolate intermediate not encountered with normal substrates. Substitution of the normal flavin with 5 deaza-FAD would thus be expected to lead to enolate formation with both normal and 3-thia-substrate analogs, because reduction of the 5-deaza-enzyme is thermodynamically highly unfavorable. However, spectrophotometric titrations show that neither ligand forms significant enolate species with the 5-deaza-FAD enzyme. Similarly, the substituted dehydrogenase catalyzes undetectable alpha proton exchange with octanoyl-CoA and ca. 1% of the corresponding rate with 3 thia-octanoyl-CoA when compared to the native enzyme. This inability to stabilize enolate species is not simply due to impaired binding of CoA-thioester analogs, because binding of a range of ligands is weakened by only 2- to 10-fold with the 5-deaza-enzyme. 4-Thia-trans-2-enoyl-CoA product is polarized normally on binding to the substituted protein, showing that this critical aspect of catalysis is apparently normal. These data, together with studies with CoA-persulfide and acetoacetyl- and p-nitrophenylacetyl-CoA, suggest that 5-deaza-FAD substitution exerts subtle, unanticipated, effects on the reductive half-reaction of the medium-chain acyl-CoA dehydrogenase. The involvement of charge-transfer interactions in the acidification of weakly acidic acyl-CoA thioesters is discussed. PMID- 11488612 TI - 2-methoxyestradiol does not inhibit superoxide dismutase. AB - It has been reported in the literature that the endogenous estrogen metabolite 2 methoxyestradiol (2-ME) inhibits both manganese and copper,zinc superoxide dismutases (Mn and Cu,Zn SODs) and that this mechanism is responsible for 2-ME's ability to kill cancer cells. In fact, as demonstrated using several SOD assays including pulse radiolysis, 2-ME does not inhibit SOD but rather interferes with the SOD assay originally used. Nevertheless, as confirmed by aconitase inactivation measurements and lactate dehydrogenase release in human leukemia HL 60 cells, 2-ME does increase superoxide production in these cells and is more toxic than its non-O-methylated precursor 2-hydroxyestradiol. Other mechanisms previously suggested in the literature may explain 2-ME's ability to increase intracellular superoxide levels in tumor cells. PMID- 11488613 TI - Quantification of intracellular metabolites in Escherichia coli K12 using liquid chromatographic-electrospray ionization tandem mass spectrometric techniques. AB - The quantitative comprehension of microbial metabolic networks is a prerequisite for an efficient rational strain improvement ("metabolic engineering"). It is therefore necessary to accurately determine the concentration of a large number of reactants (i.e., metabolites, nucleotides, cofactors) in order to understand "in vivo" reaction kinetics. Quantification of intracellular concentrations of glycolytic intermediates and nucleotides in Escherichia coli K12 using a perchloric acid extraction and an LC-ESI-MS method was achieved. Intracellular metabolites (e.g., glucose 6-phosphate, fructose 1,6-bisphosphate, 6-phospho gluconate, acetyl-CoA, adenine nucleotides) were quantified under defined (glucose-limited steady-state) growth conditions. The method was verified by comparing the intracellular metabolite concentrations measured via LC-ESI-MS with enzymatic determinations. It is thus possible to identify and quantify more than 15 intracellular metabolites in parallel with a minimal amount of sample volume. PMID- 11488614 TI - A microfluidic device for measuring cellular membrane potential. AB - Recent developments in microfluidics have enabled the design of a lab-on-a-chip system capable of measuring cellular membrane potential. The chip accesses liquid samples sequentially by sipping from a microplate through a capillary, mixes the samples with cells flowing through a microchannel, contacts the cells with potential-sensitive dyes, and reads out cellular responses using fluorescence detection. The rate of cellular uptake of membrane-permeable, ionic fluorophores by THP-1 cells was found to depend strongly on membrane potential. The ratio of the fluorescence of the anionic dye DiBAC(4)(3) and the cationic dye Syto 62 taken up by cells was found to double for every 33 mV change in membrane potential. The utility of this approach was demonstrated by assaying ion channel activity in human T lymphocytes. Because of the high sensitivity, low cellular and reagent consumption, and high data quality obtained with the microfluidic device, the lab-on-a-chip system should be widely applicable in high-throughput screening and functional genomics studies. PMID- 11488615 TI - RNA isolation and fractionation with compaction agents. AB - A new approach to the isolation of RNA from bacterial lysates employs selective precipitation by compaction agents, such as hexammine cobalt and spermidine. Using 3.5 mM hexammine cobalt, total RNA can be selectively precipitated from a cell lysate. At a concentration of 2 mM hexammine cobalt, rRNA can be fractionated from low molecular weight RNA. The resulting RNA mixture is readily resolved to pure 5S and mixed 16S/23S rRNA by nondenaturing anion-exchange chromatography. Using a second stage of precipitation at 8 mM hexammine cobalt, the low molecular weight RNA fraction can be isolated by precipitation. Compaction precipitation was also applied to the purification of an artificial stable RNA derived from Escherichia coli 5S rRNA and to the isolation of an Escherichia coli-expressed ribozyme. PMID- 11488616 TI - Expression microarray hybridization kinetics depend on length of the immobilized DNA but are independent of immobilization substrate. AB - Expression microarrays are often constructed by the immobilization of PCR products on two-dimensional modified glass slides or on three-dimensional microporous substrates. In this study we investigate whether the length of the immobilized species and the substrate choice influence hybridization dynamics. Using a simple bimolecular mass action controlled model to describe hybridization, we observed that the extent of hybridization and the initial velocities were directly dependent on the length of the immobilized species. An inflection point was noted at a length of 712 bases, above which the influence of length on hybridization rate decreased. Interestingly, we observed no differences in these parameters whether hybridization occurred on a two- or three-dimensional surface. Furthermore, the affinity of the solution phase labeled species for the immobilized species was identical for all arrayed lengths on both surfaces. These data indicate a similar interaction of the noncovalently immobilized species with either surface. Finally, we have determined that competitive hybridization on expression microarrays is nonlinear with respect to time and concentration of competitor. This observation is critical for analysis of expression array data. PMID- 11488617 TI - Binding of bovine serum albumin to heparin determined by turbidimetric titration and frontal analysis continuous capillary electrophoresis. AB - The association of proteins with glycosaminoglycans is a subject of growing interest, but few techniques exist for elucidating this interaction quantitatively. Here we demonstrate the application of capillary electrophoresis to the system of serum albumin (SA) and heparin (Hp). These two species form soluble complexes, the interaction increasing with reduction in pH and/or ionic strength (I). The acid-base property of Hp was characterized by potentiometric titration of ion-exchanged Hp. Conditions for complex formation with SA were qualitatively determined by turbidimetry, which revealed points of incipient binding (pH(c)) and phase separation (pH(phi)), both of which depend on I. At pH > pH(phi), i.e., prior to phase separation, frontal analysis continuous capillary electrophoresis was used to measure the concentration of free protein and to determine the protein-HP binding isotherm. The binding isotherms were well fit by the McGhee-von Hippel model to yield quantitative binding information in the form of intrinsic binding constants (K(obs)) and binding site size (n). The strong increase in K(obs) with decrease of pH or I could be explained on the basis of electrostatic interactions, considering the effects of protein charge heterogeneity. The value of n, independent of pH, was rationalized on the basis of size considerations. The implications of these findings for clinical applications of Hp and for its physiological behavior are discussed. PMID- 11488618 TI - Colorimetric assay method for determination of the tannin acyl hydrolase (EC 3.1.1.20) activity. AB - A new colorimetric method of tannase (tannin acyl hydrolase, EC 3.1.1.20) assay has been developed using its specific substrate tannic acid. It is based on the changes in optical density of substrate tannic acid after enzymatic reaction at 530 nm. The residual tannic acid was measured by a modified BSA precipitation method. This assay is very simple, reproducible, and very convenient, and with it tannase activity can be measured in relation to the growth of the organism. PMID- 11488619 TI - Determination of triazolam involving its hydroxy metabolites in hair shaft and hair root by reversed-phase liquid chromatography with electrospray ionization mass spectrometry and application to human hair analysis. AB - A sensitive method using reversed-phase liquid chromatography coupled with electrospray ionization mass spectrometry has been developed for simultaneous determination of triazolam and its hydroxy metabolites in hair. After the addition of deuterium-labeled 1-hydroxymethyltriazolam as an internal standard, the analytes in hair shaft and hair root samples were extracted with a basic medium, CH(2)Cl(2):MeOH:28% NH(4)OH (20:80:2) at room temperature overnight. The chromatographic separation of the analytes was achieved using a semimicro HPLC column (3-microm particle size; 100 x 2.0-mm i.d.) by gradient elution with acetonitrile in water containing 1% acetic acid as eluent. The mass spectrometer was operated in selected-ion monitoring mode at quasi-molecular ions [M+H](+) of triazolam and its metabolites. The method has been applied to determine the incorporation of triazolam and its metabolites into the hair shafts and hair roots of Dark Agouti rats administered 3 or 6 mg/kg triazolam intraperitoneally twice a day for 5 days. Triazolam, 1-hydroxymethyltriazolam, and 4 hydroxytriazolam were incorporated into the hair shafts and the hair roots. The concentration of 4-hydroxytriazolam was the highest of all compounds detected. An unknown substance considered to be 1,4-dihydroxytriazolam also appeared in the hair samples. The structural elucidation was performed with online HPLC-MS after acetylation of the substance with acetic anhydride and pyridine. The time course studies of triazolam and the metabolites in both rat hair roots and plasma were carried out after single intraperitoneal administration of triazolam. The concentrations of triazolam and the metabolites in the hair roots reflected those in the plasma. The proposed method using selected-reaction monitoring was applied to the determination of triazolam and the metabolites in human hairs of a triazolam addict. Triazolam, 1-hydroxymethyltriazolam, and 4-hydroxytriazolam were identified in the black hair shafts, whereas only triazolam was detected in the hair roots and the white hair shafts. This is the first report on the detection of triazolam and its metabolites in human hairs. PMID- 11488620 TI - A novel purification method for histidine-tagged proteins containing a thrombin cleavage site. AB - A general procedure for the purification of histidine-tagged proteins has been developed using immobilized metal-ion affinity chromatography. This two-step purification method can be used for proteins containing a hexahistidine tag and a thrombin cleavage site, yielding high amounts of purified protein. The advantage of this method is that thrombin is used instead of imidazole in the final purification step. Imidazole can influence NMR experiments, competition studies, or crystallographic trials, and the presence of imidazole often results in protein aggregates. Removal of the His-tag results in a form of the protein of interest in which no additional tags are present, resembling the native form of the protein, with only three additional amino acids at the N-terminal side. Our method is compared with a more conventional method for the purification of the Azotobacter vinelandii NIFL PAS domain, overexpressed in Escherichia coli. It also proves to be successful for three different His-tagged proteins, the Klebsiella pneumoniae NTRC protein, and the A. vinelandii NIFA and NIFL proteins, and therefore it is a general method for the purification of His-tagged proteins. PMID- 11488621 TI - Accurate determination of rate constants of very slow, tight-binding competitive inhibitors by numerical solution of differential equations, independently of precise knowledge of the enzyme concentration. AB - This paper is concerned with the determination of rate constants characterizing the binding and release of a slow binding inhibitor to and from an enzyme, here almond beta-glucosidase. We demonstrate the inability of the conventional method to yield reliable rate constants when one or more of these is less than 1 x 10( 4) per second. Instead one must use the much more accurate fitting of rate constants of the set of simultaneous differential equations characterizing the kinetic model. This procedure has the added advantage, when properly used, that the rate constants found pertaining to the inhibitor are largely insensitive to the particular value used for the enzyme concentration; i.e., the same data set may be fitted using a range of enzyme concentrations with no change in the resulting parameters. Hence the method can be used when little is known about the enzyme, except for the value of K(m), which is readily determined. Also, we report the somewhat unexpected finding that the association rate constant for the substrate (4-nitrophenyl-beta-d-glucopyranoside) is about one-third of the value of the corresponding rate constant for the inhibitor. The method is used to determine rate constants at several temperatures for the strong, slow binding inhibitor 2-phenethylglucoimidazole 1, enabling us to compute standard thermodynamic functions. The identity of these functions with those of isofagomine (2) reported earlier leads us to argue that the two compounds share a common binding mechanism, involving the same groups, whereas the different stabilities of the enzyme-inhibitor complexes must reside in those parts of the molecules that are not identical. PMID- 11488622 TI - Investigations into biochemical changes due to diurnal variation and estrus cycle in female rats using high-resolution (1)H NMR spectroscopy of urine and pattern recognition. AB - Metabonomic methods utilizing (1)H NMR spectroscopy and pattern recognition analysis (NMR-PR) have been applied to investigate biochemical variation in a control population of female rats over time in relation to diurnal and estrus cycle fluctuations. Urine samples were collected twice daily (6 AM-6 PM and 6 PM 6 AM) from female rats (n = 10) for a period of 10 days. (1)H NMR spectroscopic analysis and PR were performed on each sample. Subtle differences in the endogenous metabolite excretion profiles of urine samples at the various stages of the estrus cycle were observed. The main inherent metabolic clustering in the principal components analysis (PCA) maps was related to interrat variation and was observed in the first two principal components (PCs), accounting for 66% of the variance in these data. Separation of urinary data according to time of sampling (day and night) was achieved in the lower PCs. Some of the differences in the urinary profiles of day and night samples causing this separation were attributed to the increase in metabolic activity of the rat during the night. Individual rat data were also mapped as a function of time, using PCA, to produce a metabolic trajectory, which in a number of cases facilitated separation of one or more stages of the estrus cycle. Several of the fluctuations observed between urine samples collected during the different stages of the estrus cycle may be related to hormone levels. Although variation in metabolite profiles relating to both diurnal and hormonal variation could be detected these perturbations were minor compared with the effects observed due to interrat variation. This is the first time that a hormonal cycle has been described for individuals based on NMR spectroscopic and multivariate analysis of metabolic data and shows the value of metabonomic methods in the investigation of physiological variation and rhythms. PMID- 11488623 TI - The analysis of heparin-protein interactions using evanescent wave biosensor with regioselectively desulfated heparins as the ligands. AB - Evanescent wave biosensor has been recently employed as a powerful tool for analyses of macromolecular interactions. In the present study, evanescent wave biosensor analysis was developed to analyze the heparin-protein interaction using as ligands a series of heparin derivatives regioselectively desulfated by chemical methods, particularly to evaluate the effect of each sulfate group of heparin. The method for immobilizing heparin on the cuvette of the evanescent wave biosensor equipment was optimized to obtain the high response required for accurate measurement. The best result was achieved when the amino group introduced at the reducing end of heparin was coupled with carboxymethyl dextran on the surface of the cuvette using glycolchitosan as a multivalent linker. The established system appeared to describe well the interactions of heparin with such proteins as acidic and basic fibroblast growth factors and tissue factor pathway inhibitor. PMID- 11488624 TI - Chloroplast biogenesis 84: solubilization and partial purification of membrane bound [4-vinyl]chlorophyllide a reductase from etiolated barley leaves. AB - [4-Vinyl] chlorophyllide a reductase (4VCR) is a key enzyme of the chlorophyll (Chl) biosynthetic pathway. It catalyzes the conversion of divinyl chlorophyllide (Chlide) a to monovinyl Chlide a by reduction of the vinyl group at position 4 of the macrocycle to ethyl. 4VCR is a membrane-bound enzyme, embedded in etioplast and etiochloroplast membranes. A study of the regulation and properties of this enzyme is mandatory for a comprehensive understanding of the biosynthetic heterogeneity of Chl biosynthesis. Solubilization and partial purification of 4VCR are described for the first time. The enzyme was solubilized with 5 mM Chaps and was partially purified by chromatography on DEAE-Sephacel and Cibacron Blue 3GA-1000 agarose. An overall 20-fold purification was achieved. The partially purified enzyme was stable for several months at -80 degrees C. PMID- 11488626 TI - A microtiter plate assay for polyglutamine aggregate extension. AB - Polyglutamine (polyGln) aggregates are neuropathological markers of expanded CAG repeat disorders, and may also play a critical role in the development of these diseases. We have established a highly sensitive, fast, reproducible, and specific assay capable of monitoring aggregate-dependent deposition of polyglutamine peptides. This assay allows detailed studies on various aspects of aggregation kinetics, and also makes possible the detection and quantitation of low levels of "extension-competent" aggregates. In the simplest form of this assay, polyGln aggregates are made from chemically synthesized peptides and immobilized onto microplate wells. These wells are incubated for different times with low concentrations of a soluble biotinylated polyGln peptide. Europium streptavidin complexation of the immobilized biotin, followed by time-resolved fluorescence detection of the deposited europium, allows us to calculate the rate (fmol/h) of incorporation of polyGln peptides into polyGln aggregates. This assay will make possible basic studies on the assembly mechanism of polyGln aggregates and on critical features of the reaction, such as polyGln length dependence. The assay also will be a valuable tool for screening and characterizing anti aggregation inhibitors. It will also be useful for detection and quantitation of aggregation-competent polyGln aggregates in biological materials, which may prove to be of critical importance in understanding the disease mechanism. PMID- 11488625 TI - Screening assays for the mitochondrial permeability transition using a fluorescence multiwell plate reader. AB - Opening of permeability transition (PT) pores in the mitochondrial inner membrane causes the mitochondrial permeability transition (MPT) and leads to mitochondrial swelling, membrane depolarization, and release of intramitochondrial solutes. Here, our aim was to develop high-throughput assays using a fluorescence plate reader to screen potential inducers and blockers of the MPT. Isolated rat liver mitochondria (0.5 mg/ml) were incubated in multiwell plates with tetramethylrhodamine methyl ester (TMRM, 1 microM), a potential-indicating fluorophore, and Fluo-5N (1 microM), a low-affinity Ca(2+) indicator. Incubation led to mitochondrial polarization, as indicated by uncoupler-sensitive quenching of the red TMRM fluorescence. CaCl(2) (100 microM) addition led to ruthenium red sensitive mitochondrial Ca(2+) uptake, as indicated by green Fluo-5N fluorescence. After Ca(2+) accumulation, mitochondria depolarized, released Ca(2+) into the medium, and began to swell. This swelling was monitored as a decrease in light absorbance at 620 nm. Swelling, depolarization, and Ca(2+) release were prevented by cyclosporin A (1 microM), confirming that these events represented the MPT. Measurements of Ca(2+), mitochondrial membrane potential, and swelling could be made independently from the same wells without cross interference, and all three signals could be read from every well of a 48-well plate in about 1 min. In other experiments, mitochondria were ester-loaded with carboxydichlorofluorescein (carboxy-DCF) during the isolation procedure. Release of carboxy-DCF after PT pore opening led to an unquenching of green carboxy-DCF fluorescence occurring simultaneously with swelling. By combining measurements of carboxy-DCF release, Ca(2+) uptake, membrane potential, and swelling, MPT inducers and blockers can be distinguished from uncouplers, respiratory inhibitors, and blockers of Ca(2+) uptake. This high-throughput multiwell assay is amenable for screening panels of compounds for their ability to promote or block the MPT. PMID- 11488627 TI - The stability constants of copper(II) complexes with some alpha-amino acids in dioxan-water mixtures. AB - In this study, the overall stability constants of copper(II) complexes with some alpha-amino acids (glycine, dl-alanine, dl-valine, l-leucine, l-asparagine, l glutamine) were determined by potentiometric titration in water, 25% dioxan-75% water, 35% dioxan-65% water, 50% dioxan-50% water, and 60% dioxan-40% water. The titrations were performed at 25 degrees C, under nitrogen atmosphere, and the ionic strength of the medium was maintained at 0.10 M by using sodium perchlorate. The formation curves of their complexes (n-p[L]) were obtained by means of the titration data. Then the stability constants were determined in relation to these curves. The mol ratio of copper(II) to alpha-amino acid was also determined and it was found that the complexes were CuL(2) type. Another important result obtained was that the tendency of amino acids to form complexes with copper(II) was greater in dioxan-water mixtures compared to water. PMID- 11488628 TI - Lambda exonuclease-based subtractive hybridization approach to isolate differentially expressed genes from leaf cultures of Paulownia kawakamii. AB - Genes that are preferentially expressed in a particular developmental pathway can be isolated by subtractive hybridization (SH). We developed a PCR-based approach coupled with lambda exonuclease digestion that allows for generating single stranded tester and driver nucleic acids suitable for SH starting from cDNA libraries. An efficient subtraction strategy was developed to overcome some of the problems in the previously described SH protocols, such as the need for large amounts of experimental tissue, RNase contamination during solution hybridization, and postsubtraction recovery of nucleic acids. We used this method to obtain cDNA corresponding to genes expressed during adventitious shoot regeneration from excised leaf cultures of the fast-growing tree Paulownia kawakamii. Over 36 cDNA clones were isolated and 1 of the differentially expressed clones codes for a leucine zipper transcription factor. This clone showed about sixfold higher level of expression in the shoot-forming tissues (tester) compared to that in the callus-forming tissues (driver) of Paulownia, suggesting that differentially expressed genes can be efficiently isolated using this simple lambda exonuclease-based subtractive hybridization method. PMID- 11488629 TI - Simultaneous determination of testosterone metabolites in liver microsomes using column-switching semi-microcolumn high-performance liquid chromatography. AB - A sensitive and selective column-switching semi-microcolumn high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of testosterone and eight of its metabolites (6alpha-, 6beta-, 16alpha-, 16beta-, 7alpha-, 2alpha-, and 2beta-hydroxytestosterone, and androstenedione) in liver microsomes. After incubation for 10 min, testosterone and its metabolites were extracted from the microsomes with ethyl acetate, and the extract was evaporated to dryness. The residue was dissolved in the mobile phase and loaded onto the HPLC system. The analytes were first concentrated in a precolumn and subsequently transferred to the analytical column, where they were separated using linear gradient elution. A UV detector set at 254 nm was used to detect the analytes. This newly developed method clearly separated TES and the metabolites with high resolution and was found to be reproducible with intra- and interday variability of <10.7%. This method has been subsequently used to determine the testosterone hydroxylation activities catalyzed by 15 different recombinant CYP isozymes. The results confirmed the formation of stereoselectively hydroxylated metabolites by each CYP isozyme. PMID- 11488630 TI - Introduction of a (poly)histidine tag in L-lactate dehydrogenase produces a mixture of active and inactive molecules. AB - A (poly)histidine tag was fused to either the N- or the C-terminus of L-lactate dehydrogenase (LDH) of Bacillus stearothermophilus to facilitate purification and immobilization of these enzymes. The C-terminally tagged enzyme displayed lower activity compared both to the wild-type and to the N-terminally tagged variant. The reason for this loss of activity was investigated by affinity chromatography of the enzymes on a 5'-AMP-Sepharose resin and by size-exclusion chromatography. The C-terminally tagged enzyme could be separated into an inactive, unbound fraction and an active, bound fraction. Further differences between the C terminally tagged enzyme and the N-terminally tagged and wild-type LDH were observed on size-exclusion chromatography of the three enzymes. These data suggest that the introduction of a "his-tag" at the C-terminus may induce misfolding of the LDH and serve as a warning that the introduction of a (poly)histidine tag can produce unforseen changes in a protein. PMID- 11488631 TI - Quantitative analysis of 5-oxo-6,8,11,14-eicosatetraenoic acid by electrospray mass spectrometry using a deuterium-labeled internal standard. AB - 5-Oxo-6,8,11,14-eicosatetraenoic acid (5-oxo-ETE), a metabolite of arachidonic acid formed by the 5-lipoxygenase pathway, is a potent eosinophil chemoattractant that may be an important mediator in asthma. To further investigate the physiological and pathological roles of 5-oxo-ETE we have developed a mass spectrometric assay employing a tetradeuterated analog (5-oxo-[11,12,14,15 (2)H]ETE) as an internal standard. Collision-induced dissociation of the quasimolecular anion of 5-oxo-[11,12,14,15-(2)H]ETE (m/z 321) resulted in the formation of a major ion at m/z 207 that retained all four deuterium atoms. Measurement of the ratio of ions at m/z 203 (endogenous 5-oxo-ETE) and m/z 207 permitted quantitation of this compound by liquid chromatography-mass spectrometry-mass spectrometry using multiple reaction monitoring. The resulting assay was highly sensitive (< or =20 pg/sample) and selective, enabling detection of the amount of 5-oxo-ETE produced by as few as 10,000 neutrophils. This assay should permit measurement of 5-oxo-ETE in biological fluids, enabling evaluation of its role in asthma and other inflammatory diseases. PMID- 11488632 TI - Transient insulin autoantibody expression independent of development of diabetes: comparison of NOD and NOR strains. AB - NOD mice spontaneously develop anti-insulin autoantibodies associated with the subsequent development of diabetes. NOD mice that express insulin autoantibodies at 8 weeks of age have a diabetes risk exceeding 90%, while mice that do not express autoantibodies by 16 weeks have a risk of less than 20%. NOD female mice expressed insulin autoantibodies more often than male mice (13/15+ vs. 6/15+). Autoantibodies characteristically developed between 8 and 20 weeks and then for most mice became negative at diabetes onset in NOD mice. In the diabetes-free strain NOR mice, spontaneous expression of insulin autoantibodies was observed in less mice (female 8/15+, male 3/10+) compared to NOD mice. The expression of autoantibodies was transient in NOR mice and followed the same time-course as for NOD mice and they were all negative by 28 weeks (without progression to diabetes). No correlation was found in NOR mice between the levels of autoantibodies and insulitis. The program of insulin autoantibody expression is regulated over approximately 5 months for both NOD and NOR mice with only NOD mice developing diabetes, indicating that depending upon genetic combination, the presence of insulin autoantibodies does not always predict diabetes development. In addition, this data is not consistent with the hypothesis that the time-course of autoantibodies simply reflects the destruction of beta-cells with development of diabetes. PMID- 11488633 TI - Flexibility of TCR repertoire and permissiveness of HLA-DR3 molecules in experimental autoimmune thyroiditis in nonobese diabetic mice. AB - Experimental autoimmune thyroiditis (EAT) is inducible in genetically susceptible mice by immunization with mouse thyroglobulin (mTg). With susceptibility linked to MHC class II, EAT is useful in studying human leukocyte antigen (HLA) associations with Hashimoto's thyroiditis. In non-obese diabetic (NOD) mice, approximately 10% thyroiditis incidence occurs with aging. This potential was exploited to examine the T cell repertoire and HLA association in EAT. Similar to B10.K-Vbeta(c)mice with TCRBV genes reduced by approximately 70%, mTg-immunized NOD-Vbeta(c)mice developed thyroiditis comparable to controls, indicating plasticity of the TCR repertoire for pathogenic epitopes. HLA association was evaluated by introducing HLA-DRA/DRB1*0301 (DR3) transgene into class II-negative NOD mice (Ab(0)/NOD). Previously, this HLA-DR3 transgene rendered EAT-resistant B10.M and Ab(0)mice susceptible to both mTg- and hTg-induced EAT. These results are now confirmed. mTg-induced thyroiditis in DR3+ Ab(0)/NOD mice was comparable to that in NOD and DR3- NOD mice, and the proliferative response was stronger. By comparison, NOD mice were only moderately susceptible to hTg-induced EAT. However, thyroiditis was more severe in DR3+ Ab(0)/NOD than in DR3- NOD mice, with no difference in proliferative response to hTg harbouring heterologous epitopes. The confirmed permissiveness of HLA-DR3 molecules on an NOD background for EAT induction by both mTg and hTg supports the importance of this class II gene implicated in some patient studies. PMID- 11488634 TI - Role of MOG-stimulated Th1 type "light up" (GFP+) CD4+ T cells for the development of experimental autoimmune encephalomyelitis (EAE). AB - Experimental autoimmune encephalomyelitis (EAE) is an animal model for multiple sclerosis in humans. EAE can be passively transferred into naive syngeneic animals by administration of MOG-specific T cell clones. Lymphocytes isolated from green fluorescent protein (GFP)-transgenic (Tg) mice can light up by emitting green fluorescence, thus making it feasible to use such animals in a passive transfer model for EAE. When MOG-sensitized splenic lymphocytes from GFP Tg mice were adoptively transferred to irradiated, syngeneic C57BL/6 and RAG-1(-/ )mice, typical symptoms of EAE developed. Analysis of the reconstituted mice with EAE revealed prominent infiltration of fluorescing (GFP+), CD4+ T cells into the central nervous system (CNS). Real-time confocal imaging revealed these cells in the spinal cords and brains of recipient mice. This infiltration was also confirmed by anti-GFP monoclonal antibodies. Furthermore, quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) evaluation indicated that the infiltrating GFP+, CD4+ T cells exclusively produced T helper type 1 (Th1) cytokines, especially interferon-gamma (IFN-gamma). These results clearly show that MOG-specific CD4+ T cells preferentially invade into the CNS and mediate the development of EAE by producing Th1-biased cytokines. PMID- 11488635 TI - The effect of dose, gender, and non-H-2 genes in murine mercury-induced autoimmunity. AB - We have studied the effect of dose, treatment time, gender and non-H-2 genes on immune parameters and toxicokinetics in murine mercury-induced autoimmunity (HgAI). The partly-proven mechanism for HgAI is the modification of the autoantigen fibrillarin by mercury, followed by a T cell-dependent immune response driven by the modified fibrillarin. In the H-2 congenic (H-2(S)) mouse strains A.SW and B10.S given (203)HgCl(2) in a dose of 0.25-8 mg Hg/l drinking water for up to 10 weeks, the internal dose measured as the whole-body retention of mercury reached steady state within 5 weeks. Fifty percent of the steady state level was reached already after 2 days. Conditions therefore exist for a rapid modification of fibrillarin, followed by a T cell-dependent immune response, which is consistent with the presence of anti-fibrillarin antibodies (AFA) in serum after 2 weeks. AFA developed in a dose-dependent pattern. Serum IgE showed a dose-dependent increase with a maximum after 1-2.5 weeks followed by a distinct decline towards the baseline level. Substantial polyclonal B-cell activation (PBA) developed in the highest dose groups only. Since AFA developed using lower doses too, PBA can be excluded as a general mechanism for induction of AFA. Tissue immune-complex (IC) deposits were present in the highest dose groups only, indicating a possible causality between PBA and IC deposits. The substantially lower whole body and organ mercury level needed to induce AFA in the A.SW strain as compared with the H-2 congenic B10.S strain, demonstrates that genetic factors outside the H-2 region, and not related to toxicokinetics, modifies the autoimmune response. In contrast, the difference in mercury thresholds for induction of IgE was only slight between A.SW and B10.S mice, indicating basically different mechanisms for induction of AFA and serum IgE. PMID- 11488636 TI - Interference with CD28, CD80, CD86 or CD152 in collagen-induced arthritis. Limited role of IFN-gamma in anti-B7-mediated suppression of disease. AB - We have investigated interference with co-stimulation by administering mAbs towards CD28, CD80, CD86, and CD152 in mice immunized for the development of collagen-induced arthritis (CIA). Anti-CD80 and anti-CD86 treatment inhibited disease score and incidence, whereas anti-CD28 treatment led only to a delayed disease onset. Administration of anti-CD152 had no effect. The CII-specific Ab response was suppressed by the co-stimulatory blockade, with a stronger effect on IgG1 than on IgG2a. The CII-driven T cell proliferation, on the other hand, was not affected. Furthermore, T cells primed in the presence of either anti-B7 or anti-CD28 produced markedly increased amounts of IFN-gamma in response to CII. To investigate whether this increase in IFN-gamma was related to disease suppression, IFN-gamma-deficient mice were immunized with CII, treated with anti B7 and followed for the development of arthritis. As in the wild-type mice, administration of anti-B7 to IFN-gamma-deficient mice led to a reduced disease incidence and severity as well as reduced anti-CII IgG titers. Collectively, these data stress the importance of co-stimulation for the delivery of B cell help rather than for production of Th1 cytokines. We also demonstrate that the enhanced production of IFN-gamma observed after B7-blockade is not accountable for the anti-B7 mediated inhibition of CIA. PMID- 11488637 TI - Expression of protein tyrosine phosphatase-like molecule ICA512/IA-2 induces growth arrest in yeast cells and transfected mammalian cell lines. AB - The ICA512/IA-2 molecule, a protein with similarity to receptor-type protein tyrosine phosphatases, was discovered during studies to identify autoantigens in Type 1 diabetes. The biological function of ICA512/IA-2 is unknown. We describe striking effects of ICA512/IA-2 on viability and growth of both yeast cells and cultured mammalian cells. In transformed yeast Saccharomyces cerevisiae cells, expression of ICA512/IA-2 induced growth retardation as judged by measurements of optical density and counts of colony-forming units. In contrast, expression of the intracellular domain (amino acids 600-979) of ICA512/IA-2 in yeast or mammalian cells had no such effects. In investigations on apoptosis, expression of ICA512/IA-2 in yeast cells caused loss of plasma membrane asymmetry, but not release of cytochrome c from mitochondria which did occur in a control system after expression of the pro-apoptotic molecule Bax. Possible interactions between ICA512/IA-2 and components of the cytoskeleton were not supported by studies on staining of fixed yeast cells with phalloidin-Texas Red. With transfected mammalian cell lines COS-7 and NIH3T3, expression of ICA512/IA-2 likewise induced growth arrest, with some of the morphological features of apoptosis. Thus obligatory expression of ICA512/IA-2 in eukaryotic cells causes disruption of cellular activities, with growth arrest in yeast and nuclear pycnosis/fragmentation in mammalian cells. A possible explanation is that growth inhibition reflects a part of the presently unknown function of ICA512/IA-2. PMID- 11488638 TI - Autoimmunity to the p53 protein is a feature of systemic lupus erythematosus (SLE) related to anti-DNA antibodies. AB - The induction of anti-DNA autoantibodies in systemic lupus erythematosus (SLE) patients is problematic because mammalian DNA is poorly immunogenic at best. Here we demonstrate a chain of connected antibodies in SLE patient sera that could account for the induction of anti-DNA antibody, and possibly for some of the pathogenic features of SLE. We now report that SLE patients, in addition to anti DNA, produce antibodies to the carboxy-terminal domain of the tumour suppressor molecule p53; this p53 domain recognizes damaged DNA. Hence, these anti-p53 antibodies could mimic damaged DNA immunologically. Indeed, SLE sera do contain anti-idiotypic antibodies to a prototypic anti-p53 antibody. Moreover, SLE anti DNA antibodies also recognize this type of anti-p53 antibody. Indeed, binding of affinity-purified anti-DNA both to DNA and to the anti-p53 antibody could be blocked by a p53 peptide derived from the DNA-binding domain. This mimicry of the p53 DNA-binding domain by the SLE anti-DNA antibodies is functional: activation of the p53 molecule could be inhibited by such anti-DNA antibodies. Thus, anti DNA antibodies may arise in SLE patients by a chain of idiotypic autoimmunity centered around p53 autoimmunity. The SLE anti-DNA and anti-p53 antibodies can functionally block p53 activation, and so could affect apoptosis. PMID- 11488639 TI - Activation of natural interferon-alpha producing cells by apoptotic U937 cells combined with lupus IgG and its regulation by cytokines. AB - We recently demonstrated that IgG from patients with systemic lupus erythematosus (SLE) in combination with U937 cells made apoptotic by UV-irradiation, can induce interferon-alpha (IFN-alpha) production in normal peripheral blood mononuclear cells (PBMC). In the present study we show by flow cytometry that the actual IFN alpha producing cells (IPC) among PBMC had the same phenotype (HLA-DR+, CD4+, CD11b-, CD11c-, CD14-, CD19-, CD32-, CD36+, CD40+, CD45RA+, CD68+, CD83+, CD86-, IL-3R+ and IL-10R-) and low frequency (approximately 2/10(4)PBMC) as the IPC activated by Herpes simplex virus type I. Consequently, these cells correspond to the natural IPC, also described as type 2 precursor dendritic cells. We also demonstrated that cytokines of possible importance in the pathogenesis in SLE had effects on the IFN-alpha production. Specifically, the IFN-alpha production was strongly increased by the type I IFNs, IFN-alpha and -beta, but markedly inhibited by IL-10 and also to some extent by TFN-alpha. In contrast, the cytokines IFN-gamma, IL-6, TGF-beta and GM-CSF had no clear effects. No production of IL-10 was detected in PBMC stimulated by apoptotic U937 cells and SLE IgG. These results may explain the cause of the ongoing IFN-alpha production in SLE patients and its relation to the autoimmune process. PMID- 11488640 TI - Different dysregulations of the natural antibody repertoire in treated and untreated HIV-1 patients. AB - To investigate a possible dysregulation of the autoantibody network in AIDS patients, the relative activity of representative natural antibodies was measured in serum IgG and IgM. These immunoglobulins were purified from two cohorts of 20 HIV-infected patients undergoing, or not, a triple combination therapy. A cohort of 20 normal patients was used as a control. Marked alterations of the natural antibody repertoire were observed, varying according to the isotype and specificity of the antibody studied. For the classical self-protein antigens, human actin and myosin, the changes observed in the untreated cohort were absent in the treated cohort. In contrast, no changes, or even increased changes of the activity of antibodies to special antigens, DNA and TNP, occurred in the treated cohort. The differences were highly significant, indicating that this repertoire is regulated and not randomly modified by the disease. These results suggest the presence of different factors of dysregulation of the B cell repertoire of natural antibodies associated with the disease as well as with the treatment. These major dysregulations may favor the autoimmune phenomena observed during HIV infection. PMID- 11488641 TI - Genomic analysis of differentially expressed genes in liver and biliary epithelial cells of patients with primary biliary cirrhosis. AB - The characterization of differentially expressed genes provides a powerful tool for identifying molecules that may be involved in the pathogenesis of disease. We have used two independent techniques to identify overexpressed transcripts in bile duct cells and in liver from patients with primary biliary cirrhosis (PBC). In the first method, we used suppressive subtractive hybridization to compare mRNA from isolated PBC bile duct epithelial cells (BECs) to normal BECs and identified 71 clones as transcribed at higher levels in PBC-BECs. Amongst these clones, 62/71 had matches in a non-redundant nucleotide database and 9/71 had matches in an EST database. Of the 62 clones, 51/62 include a complexity of genes involved in cell proliferation, signal transduction, transcription regulation, RNA processing, carbohydrate metabolism and hypothetical/unknown proteins; 4/62 were identified as interstitial collagenase and collagenase precursors, 4/62 as ribosomal proteins, 3/62 as mitochondrial DNA. The mitochondrial cDNA sequences included cytochrome c oxidase, Wnt-13, and the pHL gene, a c-myc oncogene containing coxIII sequence. In the second method, we constructed cDNA libraries from three different PBC livers and sequenced a total of 12,324 independent clones. These 12,324 clones underwent virtual subtraction with 2,814,148 independent clones from Incyte LifeSeq libraries. Twenty one sequences were identified as unique to PBC liver. Collectively, these approaches identified a number of genes involved in signalling, RNA processing, mitochondrial function, inflammation, and fibrosis. Interestingly, both Wnt-13 and Notch transcripts are overexpressed in PBC liver. Further studies are needed to focus on the significance of these genes during the natural history of disease. PMID- 11488642 TI - Chronic Chagas' disease cardiomyopathy patients display an increased IFN-gamma response to Trypanosoma cruzi infection. AB - One-third of all Trypanosoma cruzi -infected patients eventually develop chronic Chagas' disease cardiomyopathy (CCC), a particularly lethal inflammatory dilated cardiomyopathy, where parasites are scarce and heart-infiltrating mononuclear cells seem to be the effectors of tissue damage. Since T. cruzi is a major inducer of interleukin-12 production, the role of inflammatory cytokines in the pathogenesis of CCC was investigated. We assayed cytokine production by peripheral blood mononuclear cells (PBMC) from CCC and asymptomatic T. cruzi infected (ASY) individuals, as well as by T cell lines from endomyocardial biopsies from CCC patients. PBMC from CCC and ASY patients produced higher IFN gamma levels than normal (N) individuals in response to B13 protein and phytohaemagglutinin PHA; IFN-gamma high responders (> or =1 ng/ml) were 2-3 fold more frequent among CCC patients than ASY individuals. Conversely, IL-4 production in response to the same stimuli was suppressed among T. cruzi infected patients. The frequency of PHA-induced IFN gammaproducing cells on PBMC was significantly higher among CCC than ASY and N individuals. IFN-gamma and TNF alpha were produced by ten out of ten PHAstimulated T cell lines from CCC patients; IL-2 and IL-10 were produced by four out of ten and one out of ten lines, respectively; IL-4, IL-1alpha, IL-1beta, IL-6 and IL-12 were undetectable. Our results suggest that CCC and ASY patients may respond differentially to the IFN-gamma-inducing stimulus provided by T. cruzi infection. Given the T(1)-type cytokine profile of heart-infiltrating T cell lines from CCC patients, the ability to mount a vigorous IFN-gamma response may play a role on the differential susceptibility to CCC development. PMID- 11488644 TI - Seizures, hormones and sexuality. AB - Sexual disorders (both hyposexuality and sexual dysfunction) are common in people with epilepsy, occurring in up to two-thirds of patients. However, characteristically, patients do not spontaneously report these problems. Nocturnal penile tumescence testing suggests that the erectile dysfunction has a neurophysiological component. The aetiology remains uncertain but is likely to be multifactorial, involving neurological, endocrine, iatrogenic, cognitive, psychiatric and psychosocial factors. Epilepsy-related factors include the age of onset/duration of epilepsy along with the seizure type and focus. In addition, seizure frequency might be relevant as successful epilepsy surgery can result in an improvement in sexual functioning despite remaining on anticonvulsant medication. Endocrine changes (raised sex hormone binding globulin and reduced free testosterone) have been reported in men with epilepsy, especially when treated with hepatic-enzyme inducing antiepileptic drugs. Studies have not been performed evaluating anticonvulsants that do not induce hepatic enzymes such as lamotrigine. The association between these endocrine changes and hyposexuality is not known. The relationship between seizures, hormones and anticonvulsant medication in women is explored, focusing on issues such as catamenial epilepsy, the menopause, hormone replacement therapy and the polycystic ovarian syndrome. Suggestions for future research and treatment issues are discussed. PMID- 11488645 TI - The assessment and management of adult patients with epilepsy--the role of general practitioners and the specialist services. AB - Recommendations have been made that all patients developing seizures should be referred to specialist services for full investigation and assessment and re referred for issues such as inadequate seizure control, consideration of drug withdrawal and for pre-conceptual counselling. Bristol area general practitioners (GPs) were sent a questionnaire to determine their referral practices for adult patients with epilepsy. Details of their current management of these patients and their requirements from the specialist services were also obtained. Questionnaires were completed and returned by 67.8% of the GPs. Most referred their patients to the neurological or neuropsychiatric services. More than 70% wanted their new patients to be assessed within 4 weeks. Approximately, half at least sometimes treated their patients before this assessment of which a third never or only rarely sought advice as to the most suitable anticonvulsant. Most (71.5%) believed they had consultations with their patients with epilepsy at least yearly; however, only 34.3% had a recall system for non-attendees. Two thirds either currently audited their practices or were willing to consider doing so, and 64.4% recognized a need for regular seminars on epilepsy. Few welcomed the introduction of joint clinics but two thirds believed co-operation cards could be useful. GPs in practices with an epilepsy nurse specialist were more supportive of the use of co-operation cards and were more likely to be involved in audit. Recommendations to improve the care provided by the Primary Health Care teams and aid communications with the specialist epilepsy services are made. PMID- 11488646 TI - The prognosis of epilepsy. AB - The cardinal question for a person developing seizures is 'What is the likelihood that they will go away?' 'Prognosis' refers to the possible outcomes of a disease and the frequency at which they can be expected to occur. Prognostic factors may include demographic features, disease-specific indicators (e.g. seizure frequency, aetiology of epilepsy) or co-morbidity. Such factors do not necessarily cause the outcome, but they are associated strongly with the outcome measured. They are distinct from risk factors--which are associated with the initial development of the disorder. Ideas about the outcome for epilepsy have been altered radically in the past century by study of its epidemiology. The prognosis for epilepsy comprises a number of measurable end-points: the prediction of recurrence after a single unprovoked seizure, the chance of remission after the diagnosis of epilepsy and the risk of premature death. PMID- 11488647 TI - Idiopathic cardiac asystole presenting as an intractable adult onset partial seizure disorder. AB - Asystole can occur during partial seizures. Conversely, asystole may produce clinical features associated semiologically with partial or secondarily generalized tonic--clonic seizures. Management is so profoundly different that accurate diagnosis is critical. We performed simultaneous scalp video electroencephalographic (EEG) and electrocardiographic (ECG) recordings in three patients who presented with an intractable seizure disorder. Habitual events were captured and reviewed. These revealed similar clinical features and temporal association with the ECG and EEG findings. Idiopathic asystole was detected as the cause in all three. All underwent emergency cardiac pacemaker implantation with resultant cessation of their paroxysmal episodes. If this diagnostic study had been delayed or not performed, all of our patients could have presented as sudden unexpected death in epilepsy (SUDEP). Early video-EEG and ECG monitoring is essential in adults with intractable seizure disorders. PMID- 11488648 TI - Infantile status epilepticus in Tunisia. Clinical, etiological and prognostic aspects. AB - This retrospective study includes 139 infants (74 girls and 65 boys) treated for status epilepticus at two University hospitals in Tunisia between 1990 and 1997. Their mean age was 11 months. The majority of seizures were generalized (74%) and lasted between 30 minutes and 1 hour (70%). The cause of status epilepticus was classified as acute symptomatic in 56, febrile in 57, remote symptomatic in nine, progressive neurologic in 10 and idiopathic in seven. Overall mortality was 15.8% and neurological sequelae were identified in 36% of the cases during the mean follow-up time of 3.5 years. The incidence of significant sequelae was a function of aetiology (especially acute symptomatic causes) and age (under 1 year of age). We conclude that the most frequent causes of infantile status epilepticus in Tunisia were fever and acute symptomatic causes. Status epilepticus among infants is an important public health problem, with significant associated mortality and morbidity. Management of status epilepticus can be improved by more rapid access to appropriate medical care. PMID- 11488649 TI - Mortality among epilepsy patients attending a tertiary referral center in a developing country. AB - Epilepsy is associated with excess mortality of two to three times in developed countries. Precise epidemiological data on mortality and cause of death are not available from India or most other developing countries. This study was carried out to estimate the mortality rates and to identify the demographic and clinical characteristics associated with mortality in a hospital based cohort of epilepsy patients. A cohort of patients enrolled in the epilepsy clinic in 1985 was followed up till 1997 (12 years). The mortality rate, demographic and clinical correlate of mortality were analysed for 246 patients (men 161, women 85) who had complete data. Cause of death was not examined in this study. Eighteen (men 15, women 3) of the 246 patients (7.3%) had died during the follow up period of 12 years. The crude death rate for the state of Kerala for the year 1990 (mid period of the study) was 5.9 per thousand population. The demographic and clinical characteristics of those who died (corresponding figures for survivors are given in brackets) were as follows: mean age 33.6 years (22.8 years), presence of abnormal neurological examination 38.9% (15.4%), mental retardation 33.3% (12.8%), abnormal CT scan 38% (21.5%). Regarding the seizure frequency at the time of enrollment and eventual mortality, there were no deaths among patients who had an Engel's seizure score of less than or equal to 4 (no seizures or nocturnal seizures only). The mortality was 5% for an Engel's score of 5 or 6 and 11% for an Engel's score greater than 6. Within the group with epilepsy, higher seizure frequency at the time of initial evaluation was associated with excess mortality. Abnormality on neurological examination, older age group and male sex were other factors that correlated with excess mortality. PMID- 11488650 TI - Juvenile myoclonic epilepsy: a clinical and sleep EEG study. AB - Juvenile myoclonic epilepsy (JME) is characterized by myoclonic jerks on awakening, generalized tonic--clonic seizures (GTCS) and is associated with absence seizures in more than one third of cases. Fifteen patients with juvenile myoclonic epilepsy were studied with regard to their clinical profile, EEG data and sleep EEG findings. There was a delay in the diagnosis of JME (mean of 3.5 years) due to various reasons. Sleep deprivation was the most common precipitating factor for triggering seizures, followed by fatigue. Routine EEGs were abnormal in 73.33% of cases only and had misleading findings in 6.66%. Sleep EEGs were abnormal in 100% of cases with generalized spikes, polyspikes and slow wave discharges. Discharge rates on sleep EEGs typically increased significantly during the transition phase (i.e. the asleep to awakening stage) and we consider this to be a specific finding in appropriate clinical setting. Sleep EEGs are a more sensitive and specific tool for the diagnosis of JME while routine awake EEGs may miss or mislead. PMID- 11488651 TI - Clinical correlations of electroencephalographic occipital epileptiform paroxysms in children. AB - A longitudinal prospective approach was used to investigate clinical correlations of interictal occipital paroxysms with or without fixation-off sensitivity (FOS). Occipital paroxysms were recorded in the electroencephalograms (EEGs) of 76 children with heterogeneous clinical conditions including seizures in 39 patients. Occipital paroxysms with FOS (42 patients ) were only fractionally more frequent than non-FOS (34 patients ) and were not specific of any clinical condition. Although present and FOS-related in all 11 children with benign childhood epilepsies with occipital paroxysms (CEOP), they were also frequently encountered in symptomatic occipital epilepsy. The differentiation of CEOP from other syndromes established on clinical grounds could also be aided by the analysis of background EEG activity that was frequently significantly more abnormal in symptomatic than CEOP. Clinical characteristics and ictal seizure semiology as well as follow-up clearly distinguish two type of idiopathic CEOP syndromes: (1) early onset type or Panayiotopoulos syndrome characterized by excellent prognosis and rare, prolonged nocturnal seizures with tonic deviations of the eyes and vomiting, and (2) late onset or Gastaut type showing a common ictal visual symptomatology, co-occurrence of migraine, diurnal complex partial seizures and less favourable EEG-clinical prognosis. PMID- 11488652 TI - Treatment of refractory partial status epilepticus with multiple subpial transection: case report. AB - Status epilepticus (SE) represents a medical emergency that annually affects 60,000--150,000 individuals in the United States. Selective neuronal loss in vulnerable areas has been pathologically demonstrated following convulsive SE primarily affecting the limbic system, thalamus and cerebellum. Morbidity in those cases that follow refractory SE (RSE) is poorly documented. There have been anecdotal reports of surgical treatment for this condition, especially secondary to brain lesions. We report a 6-year-old patient who was in RSE for 60 days, without a brain lesion documented by MRI. The patient underwent multiple subpial transection (MST) of the sensorimotor cortex, which by ictal EEG and ictal SPECT proved to be the epileptogenic zone. We conclude that MST should be considered as an alternative treatment for refractory partial SE. PMID- 11488654 TI - Review article: would eradication of Helicobacter pylori infection reduce the risk of gastric cancer? AB - This article reviews the data on the epidemiology of gastric cancer, to determine if treatment of an asymptomatic individual can be justified. It reviews retrospective and prospective case-control studies of gastric cancer in Italy and other countries. Mucosa-associated lymphoid tissue lymphoma is associated with Helicobacter pylori infection. The risk of noncardia gastric cancer is higher (4 fold or greater) in those with H. pylori infection. Although no studies have shown prevention following treatment, eradication of asymptomatic H. pylori infection in an individual in the age group 40 or lower may be expected to reduce the risk of gastric cancer. PMID- 11488655 TI - Review article: the continuing dilemma of dyspepsia. AB - This article reviews the issue of dyspepsia from various perspectives. Firstly, the current relevance of dyspepsia as a major health problem on account of its high prevalence and substantial morbidity. Secondly, the various definitions of dyspepsia and the epidemiological and pathophysiological evidence that support them, as well as subclassifications. Thirdly, the issue of dyspepsia as a clinical challenge, in the absence of well established and uniform management guidelines. Thus, the title "The continuing dilemma of dyspepsia" describes realistically the current status of this important condition. PMID- 11488656 TI - Review article: the cost of diagnosing Helicobacter pylori infection. AB - Non-invasive testing and treatment for Helicobacter pylori has been recommended for dyspeptic patients in primary care and a number of recent studies have demonstrated the cost-effectiveness of this approach. As the prevalence of H. pylori infection declines, the positive and negative predictive values of individual tests will change. Cost-effectiveness is important in determining the appropriate test in individual populations. Recent studies have shown that the stool antigen test and the urea breath test have high sensitivity and specificity in the detection of H. pylori infection before and after therapy. Cost effectiveness studies have shown that when the prevalence of H. pylori infection is low or intermediate, serological tests have relatively poor accuracy compared with the stool test or the urea breath test. In populations with low or intermediate prevalence (<60%) these tests should be preferred to ELISA serology or office-based whole-blood test or serology. This is particularly true when the prevalence of H. pylori infection is less than 30% as is seen in many developed countries. When the prevalence of H. pylori infection is high (>60%), low-cost antibody tests are cost-effective. PMID- 11488657 TI - Review Article: is Helicobacter pylori relevant in the management of reflux disease? AB - Data on the interaction of reflux disease and Helicobacter pylori infection are limited in scope and rigour, controversial and difficult to interpret. Despite this, a framework of understanding is emerging, which is consistent with known effects on gastric acid secretion. In patients with moderate to severe H. pylori induced corpus gastritis, eradication can increase substantially impaired gastric acid secretion sufficiently to precipitate reflux disease in people with pre existing sub-clinical defective gastro-oesophageal competence. By contrast, reflux disease in duodenal ulcer patients probably benefits from eradication of H. pylori. There appears to be no significant impact on reflux disease from eradication in healthy subjects or individuals whose primary problem is reflux disease. Helicobacter pylori-infected reflux disease patients respond slightly better to proton pump inhibitors. These agents cause a topographic alteration of gastritis from antrum to corpus, the clinical significance of which is controversial. Many practitioners misjudge the risks and benefits of the effects of H. pylori eradication on reflux disease. Regardless of patient diagnosis, the balance is in favour of H. pylori eradication. For those in whom reflux oesophagitis development is a defined possibility, oesophagitis is mild, easily treated and most unlikely to be associated with any major risk for development of oesophageal adenocarcinoma. PMID- 11488658 TI - Review article: Helicobacter pylori and NSAID gastropathy. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) and Helicobacter pylori are known to share a number of pathogenic mechanisms, but there is no evidence to show a significant synergic action between the two risk factors. Studies assessing this subject have differed in almost every aspect of their methodology, including the definition of a NSAID user as well as the types, doses, duration and their indications for NSAID use. They also differed in their end-points, the definition of dyspepsia and the regimes used for eradication of H. pylori. However, some conclusions may be drawn from the results of clinical trials. In H. pylori positive patients without mucosal lesions, NSAIDs may aggravate dyspeptic symptoms but, with the exception of elderly patients, they do not present a definite major risk of gastric and duodenal lesions and, above all, of ulcer correlated complications. So what recommendations can be made with regard to H. pylori eradication in patients requiring treatment with NSAIDs? The microorganism and the anti-inflammatory drugs are undoubtedly independent causes of gastric and duodenal damage. Patients taking NSAIDs who are found to have gastric or duodenal ulcers should therefore be tested for the bacterium and specifically treated, since H. pylori and NSAID-induced ulcers may be macroscopically indistinguishable. Whether asymptomatic patients taking NSAIDs should be tested and treated for H. pylori infection is still a matter of debate. PMID- 11488659 TI - Review article: Helicobacter pylori, mucosal inflammation and symptom perception- new insights into an old hypothesis. AB - The role of Helicobacter pylori and the accompanying mucosal inflammatory response in functional dyspepsia is still undefined. Human and animal studies have clearly demonstrated a link between intestinal mucosal inflammation and changes in sensory-motor function. Growing clinical and basic evidence supports the concept that a similar paradigm may occur in H. pylori-related dyspepsia. The infection may both induce gastric dysmotility and trigger neuroplastic changes in the afferent neural pathways leading to visceral hyperalgesia. A reduction of central antinociceptive control systems may also play a pathophysiological role. H. pylori eradication has provided disappointing results in terms of improvement of symptoms. This may reflect the long-term recovery of neuroplastic changes occurring in the afferent nervous system or, alternatively, the incomplete resolution of gastritis and the persistent production of inflammatory mediators by resident cells in the muscularis externa. The identification of these mechanisms may provide a better understanding of the pathophysiology of H. pylori related dyspepsia and prompt innovative therapeutic approaches. PMID- 11488660 TI - Review article: the transmission of Helicobacter pylori from stomach to stomach. AB - The mode of transmission of Helicobacter pylori is largely unknown and is a matter of circumstantial evidence and speculation rather than fact. However, the principle evidence is of two sorts: the epidemiological data, providing evidence of possible risk factors associated with transmission, and the identification of potential sources from which H. pylori could be acquired. Evidence exists for several potential sources of infection and several possible modes of transmission, and it is feasible that the transmission of H. pylori varies according to the cultural and demographic circumstances. However, the most likely recognized source for H. pylori is the human stomach, although it is not known by what route the organism is transmitted to the stomach. Evidence suggests close personal contact is important and that acquisition occurs mainly in childhood. This article reviews the evidence for the source of infection and the route of transmission of H. pylori. PMID- 11488661 TI - Review article: pre-neoplastic states of the gastric mucosa--a practical approach for the perplexed clinician. AB - The sequence leading to gastric cancer can be schematically reduced to Helicobacter pylori infection-chronic gastritis-atrophy-intestinal metaplasia dysplasia-neoplasia. Although clinicians have not yet developed a uniform approach to the treatment of gastritis (when should H. pylori infection be treated?), the entity itself is not the subject of controversy. All other lesions are still the focus of debate. There are no guidelines for the management of patients with intestinal metaplasia; pathologists are still searching for universal diagnostic criteria for atrophic gastritis; dysplasia and early neoplasia have elicited scientific diatribes between Japanese and Western pathologists. Amidst such controversies and in the absence of guidelines to regulate the management of gastric lesions, the responsibility to provide sensible clinical advice is often bestowed upon pathologists. This review discusses whether pathologists have access to sufficient evidence to provide the requested advice, and whether a consensus on the management of gastric "pre neoplastic" states is within reach. We conclude that, although many sensible and useful definitions, criteria and classifications are being generated, the final decision on how to manage the individual patient with gastric lesions will continue to be based on the communication between pathologist and clinician. PMID- 11488662 TI - Review article: should we kill or should we save Helicobacter pylori? AB - Results from epidemiological studies and therapeutic clinical trials have shown that Helicobacter pylori infection causes acute and chronic active gastritis and is the initiating factor for the majority of peptic ulcer disease. Eradication of the infection with antibiotics resolves gastritis and restores normal gastric physiology, accelerates healing of peptic ulcer disease, and virtually eliminates recurrence of duodenal ulcer disease. The infection also plays an important role in the initiation and/or progression of gastric atrophy and intestinal metaplasia, which may eventually lead to the development of distal gastric cancer. Furthermore, almost all patients with gastric MALT lymphoma are infected with H. pylori and cure of the infection leads to histological regression of the tumor and maintains the regression in over 80% of patients during long-term follow-up. Preliminary uncontrolled data from Japan show that eradication of the infection significantly reduced metachronous intestinal-type gastric cancer following initial endoscopic resection of early gastric cancer and might also prevent the progression of gastric adenoma to gastric dysplasia or gastric cancer. Although this overwhelming evidence has demonstrated that H. pylori infection is bad for humans, some have questioned the wisdom of eradicating the infection in all those infected. Their arguments are largely based on hypothesis and circumstantial evidence: 1) Less than 20% of all H. pylori infected persons will develop significant clinical consequences in their lifetime. 2) H. pylori strains are highly diverse at a genetic level and are of different virulence. 3) The antiquity of H. pylori infection in humans and their co-evolution suggests that H. pylori may be a commensal to humans. Eradication of H. pylori may remove some beneficial bacterial strains and may provoke esophageal disease or gastric cancer at the cardia. However, careful review of the literature confirms that H. pylori infection is a serious pathogen albeit in a minority of those infected. It remains for carefully designed prospective studies, rather than hypothesis to make changes in the current consensus position. PMID- 11488663 TI - Buckwheat allergy. PMID- 11488664 TI - Avoidance of allergens and air pollutants in respiratory allergy. PMID- 11488665 TI - Allergen cross-reactivity between house-dust mites and other invertebrates. PMID- 11488666 TI - Skin reactions to histamine of healthy subjects after hypnotically induced emotions of sadness, anger, and happiness. AB - BACKGROUND: The severity of symptoms in asthma and other hypersensitivity-related disorders has been associated with changes in mood but little is known about the mechanisms possibly mediating such a relationship. The purpose of this study was to examine the influence of mood on skin reactivity to histamine by comparing the effects of hypnotically induced emotions on flare and wheal reactions to cutaneous histamine prick tests. METHODS: Fifteen highly hypnotically susceptible volunteers had their cutaneous reactivity to histamine measured before hypnosis at 1, 2, 3, 4, 5, 10, and 15 min after the histamine prick. These measurements were repeated under three hypnotically induced emotions of sadness, anger, and happiness presented in a counterbalanced order. Skin reactions were measured as change in histamine flare and wheal area in mm2 per minute. RESULTS: The increase in flare reaction in the time interval from 1 to 3 min during happiness and anger was significantly smaller than flare reactions during sadness (P<0.05). No effect of emotion was found for wheal reactions. Hypnotic susceptibility scores were associated with increased flare reactions at baseline (r=0.56; P<0.05) and during the condition of happiness (r=0.56; P<0.05). CONCLUSION: Our results agree with previous studies showing mood to be a predictor of cutaneous immediate-type hypersensitivity and histamine skin reactions. The results are also in concordance with earlier findings of an association between hypnotic susceptibility and increased reactivity to an allergen. PMID- 11488667 TI - The effect of vaccination with DNA encoding murine T-cell epitopes on the Der p 1 and 2 induced immunoglobulin E synthesis. AB - BACKGROUND: Immunization with naked plasmid DNA leads to strong and persistent cell-mediated and humoral immune response to plasmid encoded antigen. Vaccination of DNA encoded whole allergen has been tried, but little information is currently available on the efficacy of DNA encoding T-cell epitopes in allergic disease. The purpose of this study was to determine whether the vaccination of naked plasmid DNA encoding only T-cell epitopes suppresses the allergic reaction as effectively as naked DNA encoding whole segments of allergen. METHODS: We immunized mice with a mixed naked plasmid DNA encoding the five classes of murine T-cell epitopes on Der p 1 and Der p 2 three times at weekly intervals via an intramuscular injection of BALB/c mice. Control mice were injected with the pcDNA 3.1 blank vector. After 3 weeks, the mice were actively sensitized twice and allowed to inhale the Der p extracts intranasally six times at weekly intervals. RESULTS: The vaccinated mice showed a significant attenuated induction of Der p specific immunoglobulin E synthesis compared to controls. In terms of the Der p specific IgG2a antibody response, the vaccinated mice showed more prominent responses than the control mice group. In addition, analysis of the cytokine profile after Der p stimulation of the lymph-node cells revealed that the level of the mRNA expression of the interferon-gamma gene was higher in the vaccinated mice than in the controls. Histologic studies showed a much reduced infiltration of inflammatory cells in lung tissue of the gene-vaccinated mice in comparison with the controls. CONCLUSIONS: These results suggest that vaccination with DNA encoding T-cell epitopes effectively inhibits allergen-induced IgE synthesis and reduces cell infiltration in lung tissue. Thus, gene therapy using T-cell epitope encoding DNA presents an ideal way of combating allergic disease in the future. PMID- 11488668 TI - Genetic susceptibility to asthma and atopy among Chinese in Singapore--linkage to markers on chromosome 5q31-33. AB - BACKGROUND: Asthma and atopy are complex genetic traits, influenced by the interaction of multiple genes and environmental factors. Linkage of these traits to chromosome 5q31-33 has been shown in other populations, but has not been well studied in the Chinese. We studied linkage between asthma and atopy with markers on chromosome 5q31-33 in the Singapore Chinese. This region contains many candidate genes, including the cytokine gene cluster. METHODS: We recruited 88 Chinese families with at least two affected offspring, totaling 373 subjects, with 125 and 119 sib-pairs for atopy and asthma, respectively. All individuals were genotyped with 19 polymorphic microsatellite markers spanning a distance of 41 cM along chromosome 5q31-33. Affected sib-pair and multipoint linkage analysis was performed. RESULTS: There was evidence for linkage of the asthma and atopy phenotypes with three markers, D5S2110, D5S2011, and D5S412 (P values of 0.001 to 0.00001). Multipoint analysis further substantiated this (nonparametric linkage scores of 1.8-2.9). These findings suggest that susceptibility genes for asthma and atopy are found in this region in the Chinese. CONCLUSION: This study has shown linkage of atopy and asthma to chromosome 5q31-33 in a heterogeneous Chinese population. These findings further substantiate the notion that chromosome 5q31-33 contains "universally" important susceptibility genes for these traits. PMID- 11488669 TI - Chicken serum albumin (Gal d 5*) is a partially heat-labile inhalant and food allergen implicated in the bird-egg syndrome. AB - BACKGROUND: Chicken serum albumin (alpha-livetin) has been implicated as the causative allergen of the bird-egg syndrome. However, the clinical relevance of sensitization to this allergen has not been confirmed by specific challenge tests and environmental sampling. We investigated whether chicken albumin can be detected in air samples collected in a home with birds, and whether sensitization to this protein may cause respiratory and food allergy symptoms. The heat resistance of chicken albumin and the possible cross-reactivity with conalbumin were also investigated. METHODS: We studied eight patients with food allergy to egg yolk who also suffered from respiratory symptoms (rhinitis and/or asthma) caused by exposure to birds. Sensitization to egg yolk and bird antigens was investigated by skin and serologic tests. Hypersensitivity to chicken albumin was confirmed by specific bronchial, conjunctival, and oral provocation tests. RESULTS: All patients had positive skin tests and serum IgE against egg yolk, chicken serum, chicken meat, bird feathers, and chicken albumin. The presence of airborne chicken albumin in the domestic environment was confirmed. Specific bronchial challenge to chicken albumin elicited early asthmatic responses in six patients with asthma. An oral challenge with chicken albumin provoked digestive and systemic allergic symptoms in the two patients challenged. IgE reactivity to chicken albumin was reduced by 88% after heating at 90 degrees C for 30 min. ELISA inhibition demonstrated only partial cross-reactivity between chicken albumin and conalbumin. CONCLUSION: Chicken albumin (Gal d 5) is a partially heat labile allergen that may cause both respiratory and food-allergy symptoms in patients with the bird-egg syndrome. PMID- 11488670 TI - Three cases of childhood nocturnal asthma due to buckwheat allergy. AB - BACKGROUND: Buckwheat flour (BF) is known as a potent food allergen. Sensitization to it usually occurs by ingestion but also by inhalation in occupational or domestic exposure. Buckwheat chaff-stuffed pillows (BCP) can be contaminated with BF during the process of pilling, and many Korean children and adults use BCP for health reasons. METHODS AND RESULTS: We here present three cases of BF allergy in children using BCP, who had been treated as nonatopic asthmatics after undergoing the routine allergy skin tests and serologic tests. We took careful clinical histories, and performed skin prick tests (SPT), the radioimmunoassay (RIA) for specific IgE, the BCP-elimination test, the BF bronchial provocation test, and IgE Western blot. All three children showed positive skin reactions to BF, but none of them had positive reactions to house dust mites. Nocturnal asthmatic symptoms were improved during 7 days of BCP elimination, and asthmatic responses were observed by bronchial provocation tests with homemade BF extract. Serum BF-specific IgE antibodies and several IgE binding proteins were detected by RIA and Western blot analysis, respectively. CONCLUSIONS: Thus, a small amount of BF attached to BCP can induce BF sensitization, and BCP should be considered a main cause of childhood nocturnal asthma in those asthmatics exposed to these pillows. PMID- 11488671 TI - Molecular analysis of T-cell clonality with concomitant specific T-cell proliferation in vitro in nickel-allergic individuals. AB - BACKGROUND: The peripheral blood mononuclear cells (PBMC) of individuals with nickel contact allergy are reported to proliferate to a varying degree upon nickel stimulation in vitro. Different phenotypes of the T cells involved are described. With regard to preferential use of the T-cell receptor (TCR), analysis of the several families of the TCR-gamma gene allows rearrangement evaluation of all T cells regardless of predominant surface expression of TCR alpha/beta. METHODS: The PBMC of 10 nickel-allergic and five nonallergic individuals were cultured for 4 days in the presence of either medium, PHA, NiSO4, or tetanus toxoid (TT). Proliferation was measured by radioactive thymidine uptake and expressed as stimulation index (SI). T-cell clonality was assessed by analysis of the TCR-gamma chain gene, including the use of PCR with a primer combination covering the four main groups (Vgamma1-8, Vgamma9, Vgamma10, and Vgamma11) of the variable region of the TCR-beta chain gene. RESULTS: In the allergic individuals, proliferation to NiSO4 was significantly (P<0.05) higher than in nonallergics (mean SI: 18.05/17.87 vs 0.67/2.27). In unstimulated and PHA-stimulated cultures, there was a random TCR spectrum in both groups. In contrast, in nickel-allergic individuals or individuals with recent TT-booster, oligoclonality could be observed in the correspondingly stimulated cultures. CONCLUSION: In addition to proliferation assay, analysis of T-cell clonality may be a further means to characterize clinical hypersensitivity reactions on the basis of antigen dependent oligoclonal T-cell expansion, as in the case of tissue-infiltrating lymphocytes. PMID- 11488672 TI - Dispersion of horse allergen in the ambient air, detected with sandwich ELISA. AB - BACKGROUND: The objective was to establish an ELISA to detect horse allergen in ambient air and settled dust. METHODS: Monoclonal antibodies (mAbs) were produced against extracts of horse antigen. Two mAbs were selected and used in a sandwich ELISA. By the aid of portable pumps, air samples were collected in one stable and in the ambient air surrounding this stable. Furthermore, settled dust was collected by wiping spots with pieces of fabric, at sites within 500 m of the stable. RESULTS: Extracts of horsehair could be extensively diluted and still be positive. Extracts of cat and dog allergen failed to be detected. Furthermore, the mAbs were shown to detect an IgE-binding component. This was demonstrated by an ELISA using mAbs as capture antibody and sera from horse-allergic subjects as secondary antibody with readout depending on anti-IgE antibody. The sera with the highest RAST class to horse were positive in this ELISA. Airborne levels of horse allergen were over 500-fold higher in the stable than just outside the stable and over 3000-fold higher than at a residential building located only 12 m from the stable. Similarly, an inverse correlation was found between the distance to the stable and levels of "outdoor settled" horse allergen (r=-0.9, P<0.001). CONCLUSION: We have developed a sensitive, horse-allergen-specific, mAb assay allowing detection of low levels of horse allergens. Raised levels of horse allergen were found outdoors only in the close vicinity of the stable. PMID- 11488673 TI - No linkage and association of atopy to chromosome 16 including the interleukin-4 receptor gene. AB - BACKGROUND: Several susceptibility genes for atopy have been suggested in recent years. Few have been investigated as intensively as the interleukin-4-receptor alpha (IL4Ralpha) gene on chromosome 16. The results remain in dispute. Therefore, in a robust design, we tested for association of type I allergy to the IL4R variations I50V and Q576R, and investigated chromosome 16 for atopy candidate regions in general. METHODS: We identified 100 Danish allergy sib-pair families. Five conservative phenotypes for type I allergy were defined and evaluated. The IL4R variations were genotyped in trios and evaluated by the transmission disequilibrium test (TDT). Multipoint linkage analysis and exclusion mapping were conducted with sib-pairs analyzed for 17 microsatellite markers. RESULTS: No evidence for association or linkage to the IL4R polymorphisms was found (P values: 0.12-0.90). Linkage analysis did not support linkage of any of the phenotypes to chromosome 16. Major parts of chromosome 16 were excluded as candidate regions harboring oligogenes for type I allergy. CONCLUSION: We found chromosome 16 unlikely to harbor strong candidate genes for type I allergy. The role of the IL4Ralpha gene in the inheritance of atopy was insignificant in the Danish population. The use of conservative allergy phenotypes in the search for genes predisposing to atopic disease was discussed. PMID- 11488674 TI - Urine eosinophil protein X (EPX) is an in vitro parameter of inflammation in atopic dermatitis of the adult age. AB - BACKGROUND: Eosinophils are important effector cells in several atopic diseases. The levels of eosinophil granule-derived mediators (ECP, EPX) in serum and body fluids have been proven to be correlated with disease activity in atopic respiratory diseases and atopic dermatitis. The study aimed to demonstrate an interrelationship between urine EPX and disease activity in adult patients with atopic dermatitis. METHODS: We determined urine EPX concentration, serum ECP concentration, and peripheral blood eosinophil count in 40 adult patients with mild to severe atopic dermatitis and compared it with the disease activity as assessed with the SCORAD index. RESULTS: Urine EPX and serum ECP concentrations were significantly higher in patients with severe atopic dermatitis than in patients with mild or moderate disease (median values 123.5 vs 78.3 microg/mmol creatinine, P<0.0001; 25.4 vs 14.9 microg/l, P<0.0001, respectively). We found a significant correlation between urine EPX levels, serum ECP levels, and the SCORAD (r=0.36, P<0.0001 and 0.34, P<0.0001, respectively). CONCLUSION: Urine EPX is a useful in vitro parameter of inflammation in atopic dermatitis of the adult age. PMID- 11488675 TI - Conjunctival allergic contact hypersensitivity. PMID- 11488676 TI - Allergy to two drugs in a patient. PMID- 11488677 TI - Allergen-specific immunotherapy to mosquitoes. PMID- 11488678 TI - Asthma in pulmonary tuberculosis. PMID- 11488679 TI - Rofecoxib as an alternative in aspirin hypersensitivity. PMID- 11488680 TI - Cephalexin tolerated despite delayed aminopenicillin reactions. PMID- 11488681 TI - Urticaria to methylprednisolone sodium hemisuccinate. PMID- 11488682 TI - Circadian rhythm of U-EPX. PMID- 11488683 TI - Soy anaphylaxis. PMID- 11488684 TI - Recombinant spiked allergen extract. PMID- 11488685 TI - Contact cross-sensitization among quinolines. PMID- 11488686 TI - Anaphylaxis to beef in penicillin-allergic patient. PMID- 11488688 TI - Increased SPT reaction in fatty liver. PMID- 11488687 TI - Allergy to lipopolysaccharide? PMID- 11488689 TI - Anaphylaxis on skin exposure to grass. PMID- 11488690 TI - Measles and atopy--rash on rash? PMID- 11488691 TI - Allergy to calcitonin. PMID- 11488692 TI - Urticaria to hydrocortisone. PMID- 11488693 TI - Meloxicam in hypersensitivity to NSAIDs. PMID- 11488694 TI - Anaphylaxis to Japanese encephalitis vaccine. PMID- 11488695 TI - Hypersensitivity to IFN-alpha. PMID- 11488696 TI - Sensitization to synthetic yarns. PMID- 11488697 TI - The promotion of health careers to high school students in the New England health area: the views of high school careers advisers. AB - One way to impact positively on the shortage of health professionals in rural areas is to effectively promote health careers to rural high school students. Rural high school careers advisers play a pivotal role in this. In order to assess how rural health careers advisers working in the north-west of New South Wales currently promote health careers to their students, the New England Area Rural Training Unit carried out a survey of the area's high school careers advisers. Of the 47 high school careers advisers, 38 returned completed questionnaires, yielding a response rate of 81%. While only about one-third of careers advisers use visits by undergraduate students enrolled in tertiary health courses (42%), visits by locally practising health professionals (39%) and/or health careers site visits (27%), all careers advisers consider such promotional activities to be most effective. Improved exposure to such effective health career promotional activities for the area's high school, increasing collaboration between careers advisers and health professionals, as well as renewed efforts to identify and to foster interested students prior to Year 10, should lead to an increasing number of rural high school students enrolling in tertiary health courses. PMID- 11488698 TI - Health issues among Filipino women in remote Queensland. AB - This paper discusses health issues among Filipinas (women born in the Philippines) living in remote and rural environments in Queensland. The sample was recruited as part of the University of Queensland component of the Australian Longitudinal Study of Women's Health (ALSWH). Most of the women lived in an urban or semi-urban area (391), whereas 90 lived in rural or remote areas. Community perceptions had a much greater impact on health service utilisation in rural and remote areas than in urban areas. The transition between newcomer and old-timer is more difficult for Filipinas than for other rural women because they are visibly different from other members of the community and suffer from stigmatisation associated with perceptions of Filipinas as mail order brides. For these women, concerns about confidentiality and a reluctance to ask for support are major barriers to health service utilisation. The area of greatest concern is mental health, particularly in relation to parenting issues. Improving access may involve providing a greater awareness of what services are available and allowing women to access services in a way that does not require them to label themselves or be labelled by others. PMID- 11488699 TI - Fostering hope in people living with AIDS in Africa: the role of primary health care workers. AB - Today the medical literature is dominated by discussions on issues related to HIV/AIDS. This is not surprising considering the fact that in the history of humankind, the HIV/AIDS scenario has posed one of the greatest challenges. The reality of the physical, socioeconomic and psychological problems associated with the AIDS epidemic has become obvious to the general populace in Africa. Currently, both the AIDS victims and several others in the society continue to entertain the fear of dying from AIDS. The situation has become a source of concern to almost everyone, including primary health-care (PHC) workers. While several options are being examined to address the AIDS problem in Africa, one area that is often neglected is how to foster hope in people living with AIDS (PLWA) and their caregivers. In an attempt to examine this issue, this paper discusses the concept of hope, the cultural construct of HIV/AIDS in African countries and the role of PHC workers in fostering hope in PLWA. The paper concludes that by assisting the PLWA to develop a good sense of hope, PHC workers will be able to meet an important challenge: how to improve the quality of life for PLWA. PMID- 11488700 TI - High use of complementary therapies in a New South Wales rural community. AB - In this study a postal questionnaire was used to investigate the use of complementary therapies (CT) in a rural region of New South Wales (NSW). A total of 70.3% of respondents reported using one or more therapies with 62.7% having visited a complementary practitioner. Vitamin/mineral therapy (68.7%), chiropractic (26.1%) and massage therapy (25.1%) were the most frequently used therapies, with the most commonly visited practitioners being chiropractors (55.3%) and those trained in vitamin/mineral therapy (47.9%). Of the vitamins, vitamin C and multivitamins were used most often, with garlic and Echinacea being the most used herbal products. The main source of information were friends (64.5%) with most people indicating that CT could improve quality of life and provide relief of symptoms. Results suggest that CT use in rural communities is high and that, in light of these findings, there is a need for training in CT to be included in the undergraduate and continuing education of health professionals. Furthermore, these professionals should be alert to the potential for increased numbers of herb-drug interactions in rural communities. PMID- 11488701 TI - Insights obtained from an evaluation of a falls prevention program set in a rural hospital. AB - An evaluation of a Falls Prevention Program that took place in a 29-bed rural hospital in New South Wales is described. The aim of the project was to ascertain the overall effectiveness of the Program and to explore the usefulness of the assessment criteria in predicting falls. The sample consisted of 111 participants, representing all patients 65 years and over who were admitted to the general ward of the hospital between January and December 1997. The Falls Prevention Program had reduced the incidence of falls and was found to be effective for those patients requiring minimal assistance with walking. However, it was less effective for those using pick-up frames or forearm support frames. The patients who fell were more likely to be in the high risk category and it was concluded that while the assessment criteria was useful in predicting falls, the Falls Prevention Program could only limit the number of falls but not prevent them altogether. Age, mental status and mobility of patients in combination with time and location of falls suggested a pattern that was possibly peculiar to this rural hospital, which has implications for funding and staffing. PMID- 11488702 TI - Rural training and the state of rural health services: effect of rural background on the perception and attitude of first-year medical students at the university of melbourne. AB - The aim of this project is to investigate the relationship between medical students' background and their perception of the state of rural health services; willingness to undertake internship training or work as a doctor in a rural hospital; expected benefits and disadvantages of training or working as a doctor in a rural hospital; and factors interfering with acceptance of a job as a doctor in rural areas. A questionnaire-based survey was distributed to 100 first-year medical students attending the Faculty of Medicine, Dentistry and Health Sciences, The University of Melbourne at the end of semester 1. The response rate was 97%, including 44 males and 53 females. A strong relationship was found between rural background and an intention to undertake internship training in a rural hospital (86% of students from a rural background expressed this desire vs 30% of students from an urban background). Furthermore, all students from a rural background expressed a desire to work as a doctor in a rural hospital after completing postgraduate training. Compared to urban students, students from a rural background showed a more positive attitude towards health services in rural areas including public hospitals ( P = 0.02), private general practice ( P = 0.004), ambulance service ( P = 0.0002) and baby health centres ( P = 0.005). Citizenship or gender was not significantly related to the perception of any of these services. The ranking of factors interfering with acceptance of a job as a doctor in rural areas were different for rural and urban students. Students from rural backgrounds reported spouse/partner needs (76% vs 49%, P = 0.038) and school availability for children (59% vs 30%, P = 0.023) as barriers more frequently than urban students, respectively). On the other hand, urban students rated the following factors higher: personal factors (76% vs 53%, respectively), education opportunities (56% vs 24%), social/cultural facilities (50% vs 41%) and the need for frequent travel (29% vs 12%). None of these interfering factors were significantly different. Urban students were more likely than rural students to report that their views were a result of adverse media reports. In conclusion, students from a rural background were more willing to be trained or to work as doctors in rural areas. This was associated with a greater adverse influence by the media upon students. PMID- 11488703 TI - Costs of mobile screening for diabetic retinopathy: a practical framework for rural populations. AB - Australia's rural and remote residents experience considerably higher hospitalisation and death rates due to diabetes than their metropolitan counterparts. There is clearly a need for improved diabetes care services in these areas and interventions that target conditions associated with diabetes will yield beneficial results for the community. All people with diabetes are at risk for diabetic retinopathy, which can cause vision loss and blindness. Although vision loss and blindness due to diabetes is nearly 100% preventable through regular eye examinations, 35% of Victoria's rural population with diabetes do not have their eyes examined on a regular basis. A pilot, mobile screening program for the early detection of diabetic eye disease was conducted in rural Victoria and proved to be a successful model of adjunct eye care for people with diabetes. Actual costs from the pilot screening were applied to a permanent model for rural eye care. At A$41 per participant, costs for mobile screening were competitive with Medicare rebate costs for eye examinations. The model addresses barriers of accessibility and availability, targets a portion of the rural population with diabetes that is not otherwise having eye examinations, and is cost-saving to the Government. PMID- 11488704 TI - Developing better casemix education for rural New South Wales. AB - Casemix is now an important mechanism for the planning, evaluation and funding of health services in Australia. In New South Wales (NSW) it was believed that while staff from most hospitals in metropolitan Sydney had become both literate and vocal about casemix, staff from rural areas were less familiar and much less likely to participate in casemix initiatives. In conjunction with the NSW Casemix Clinical Committee (NCCC), NSW Health considered a special program of casemix education for rural NSW. Before an education program was attempted, NSW Health inquired into the specific needs for casemix education in rural NSW. Qualitative and quantitative methods of analysis were used. Results of the quantitative analysis indicate that the understanding of casemix classifications is highest among managers. Of concern were the relatively low proportion of Allied Health staff who had more than a vague understanding of the Sub- and Non-Acute Patient (SNAP) classification; the lack of any knowledge of the Mental Health Costing And Service Classification (MH-CASC) by nursing staff; and the lack of any knowledge of the emergency department classification: Urgency, Disposition and Age-related Groups (UDAG), either by clinical or nursing staff. The results of the qualitative analysis show that casemix education for rural areas needs to differ from metropolitan education programs. The analysis also highlights the perception of casemix in rural areas and the special circumstances in rural hospitals that place limits on the ability to use casemix more fully. PMID- 11488705 TI - Rural nurses: have we bureaucratised ourselves out of the race for collaborative practice? PMID- 11488706 TI - Menopause and the skin. AB - Women live one-third of their lives in the post-menopausal state. Significant hormonal alterations occur at the time of menopause, leading to a range of physiological disorders affecting multiple organ systems in the body. The effects of menopause on the skin have been underresearched. Many skin changes occur at the time of menopause and the cutaneous effects of hormone replacement therapy are significant. Menopausal changes in hormones may alter the biomechanical properties of the skin and certain disorders are more common in menopausal women, such as lichen sclerosus, atrophic vulvovaginitis, flushing and dysaesthetic vulvodynia. Hair and oral changes may also be associated. As the average life expectancy increases, dermatologists need to be familiar with skin diseases affecting women in this age group. PMID- 11488707 TI - Effects of low-dose ultraviolet radiation on in vivo human cutaneous recall responses. AB - Relatively few studies have examined the effects of low-dose ultraviolet (UV) radiation on in vivo human cutaneous immunity, or the ability of sunscreens to prevent UV-induced immunosuppression. We have studied the effects of solar simulated UV radiation on nickel contact hypersensitivity (CHS) in nickel allergic volunteers, and on delayed type hypersensitivity responses in Mantoux positive volunteers. Nickel CHS and Mantoux responses were significantly suppressed by acute, suberythemal UV exposures equivalent to less than 8 min summer sunlight. Both UVA and UVB wavebands were immunosuppressive, but UVA induced immunosuppression was transient, whereas UVB had a more sustained effect. Dose-responses for UV immunosuppression were determined using the nickel method, enabling calculation of in vivo sunscreen immune protection factors in a manner analogous with sun protection factor measurement. Sunscreens were found to confer significantly less protection against UV-induced immunosuppression than against UV-induced erythema. PMID- 11488708 TI - Keratoacanthoma with perineural invasion: a report of 40 cases. AB - Keratoacanthoma is a unique clinicopathological entity, despite a recent trend to regard it as a variant of squamous cell carcinoma. The occurrence of perineural invasion is an uncommon phenomenon in keratoacanthomas, with a predilection for lesions on the face. We studied a series of 40 cases of keratoacanthoma in which perineural invasion occurred. Of the 40 cases, 27 were from the head or neck region. We found no metastasis or direct death attributable to the presence of perineural invasion in the 35 cases in our series for whom follow-up data were available. In only one case did local recurrence occur and this was not considered by the authors to be directly attributable to the presence of perineural invasion. These findings add further support to the notion that keratoacanthoma is biologically different from squamous cell carcinoma. PMID- 11488709 TI - Comparative study of pemphigus vulgaris and pemphigus foliaceus in Singapore. AB - This is a retrospective study of all patients diagnosed to have pemphigus in our centre over a 3 year period. The case records of all patients with pemphigus from January 1995 to December 1997 were analysed. Fifty patients were diagnosed to have pemphigus during the study period. The diagnoses were pemphigus vulgaris in 31 patients, pemphigus foliaceus in 16, paraneoplastic pemphigus in two and IgA pemphigus in one. The average titre of anti-intercellular antibodies in patients with pemphigus vulgaris (1:96) was higher than the titre in patients with pemphigus foliaceus (1:69). The average initial dose of prednisolone required for disease control in patients with pemphigus vulgaris (62 mg/day) was significantly higher than that required for patients with pemphigus foliaceus (44 mg/day). In our study population, pemphigus vulgaris is a more severe and chronic disease than pemphigus foliaceus, as reflected in the higher titre of anti-intercellular antibodies, higher dose of systemic corticosteroids required for control of the disease, the longer duration to achieve complete remission and longer follow-up period. PMID- 11488710 TI - Frictional amyloidosis: a study of 10 cases. AB - Ten patients with macular amyloidosis were studied with particular reference to the role of friction. All 10 patients had a history of prolonged rubbing over a period of 2-5 years with various objects, such as bath sponges, brushes, towels, plant sticks and leaves. The presence of amyloid was confirmed by histochemical stains in six cases and by electron microscopy in four cases. The study confirms the role of friction in the causation of macular amyloidosis and hence, the term 'frictional amyloidosis' aptly describes the condition. The study also emphasizes the need for electron microscopy in the diagnosis of frictional amyloidosis. PMID- 11488711 TI - Spironolactone-induced hepatitis. AB - There is widespread use of spironolactone in medical practice and the indications for its use well established and side effect profile well known. Herein, we present a case of drug-induced hepatitis occurring in a 50-year-old woman using spironolactone for the treatment of androgenetic alopecia. Six weeks after commencement of spironolactone the patient became unwell, complained of an extensive itch, but no icterus or jaundice. Liver function tests found abnormally elevated bilirubin and enzymes levels. After withdrawal of spironolactone, the patient's symptoms resolved and liver function improved. To date, there has only been one other report of spironolactone-induced hepatitis. PMID- 11488712 TI - Subcutaneous panniculitic T-cell lymphoma and cytophagic histiocytic panniculitis. AB - A 43-year-old Maori man presented with a 1 month history of malaise, weight loss, anorexia, arthralgia, recurrent fever and tender erythematous subcutaneous skin lesions. Histological examination of an incisional biopsy of a lesion revealed a lobular panniculitis with an inflammatory infiltrate of atypical lymphocytes and evidence of cytophagocytosis consistent with a diagnosis of subcutaneous T-cell lymphoma. The systemic symptoms and skin lesions resolved spontaneously within 3 weeks, only to recur 2 months later, requiring treatment with oral prednisolone. T-cell gene rearrangement studies demonstrated a monoclonal T-cell receptor (gamma-chain) gene rearrangement, further supporting the diagnosis of subcutaneous panniculitic T-cell lymphoma. Treatment with chemotherapy (cyclophosphamide, doxorubicin, vincristine and prednisone) led to remission of symptoms. Four months after completing chemotherapy, the patient remained asymptomatic with a few indurated subcutaneous plaques on the chest. Biopsy of these areas revealed lobular panniculitis, lymphocytic infiltrate without cytological atypia, abundant lipophages and fibrosis and sclerosis consistent with a healing response. He remains well 24 months following chemotherapy. PMID- 11488713 TI - Lichen planus-like eruption following autologous bone marrow transplantation for chronic myeloid leukaemia. AB - A 47-year-old female treated with an autologous bone marrow transplant and cytotoxic chemotherapy developed a lichen planus-like eruption 12 months later. It involved the skin and oral mucosa, with the histological features of a lichenoid graft-versus-host reaction, including satellite cell necrosis. This eruption developed de novo. The eruption resolved with topical betamethasone valerate 0.1% cream despite the ongoing use of the immunomodulatory agent interferon-alpha2b. Such a reaction in an autologous setting has only been described once previously. PMID- 11488714 TI - A case of severe actinic prurigo successfully treated with thalidomide. AB - Actinic prurigo is an uncommon and usually persistent idiopathic photodermatosis with typical human leukocyte antigen (HLA) associations (HLA-DR4, particularly subtypes DRB1*0407 and DRB1*0401). Although its mechanism of action is not clearly understood, thalidomide has been shown to be particularly efficacious in treating actinic prurigo, among other conditions. A 31-year-old Australian woman who had suffered actinic prurigo for most of her life was treated with two courses of thalidomide (50-100 mg nocte) over consecutive summers. Remission was observed after cessation of the second course of thalidomide and had continued 4 years later. Abnormalities in the cutaneous response to ultraviolet radiation at the time of diagnosis, detected by monochromator phototesting, reverted to normal following treatment. PMID- 11488715 TI - Drug-induced linear IgA bullous disease following antibiotics. AB - A 69-year-old woman presented with pneumonia and subacute bacterial endocarditis. Nine days after intravenous vancomycin and ciprofloxacin were commenced, the patient developed a bullous mucocutaneous eruption. Clinical presentation and histopathology were consistent with drug-induced linear IgA bullous disease (LABD). The patient's lesions resolved with cessation of antibiotics. A review of the features of drug-induced LABD and the drugs that have been implicated are presented. PMID- 11488716 TI - Neutrophilic fixed drug eruption. AB - A 49-year-old man presented with a reproducible, localized amoxycillin-clavulanic acid-induced eruption. The histopathology from lesional skin revealed a neutrophilic dermatosis. These histological findings have not been reported in previous fixed drug eruptions. A brief review is undertaken comparing fixed drug eruption and the group of neutrophilic dermatoses with our case presentation. We propose a new entity of neutrophilic fixed drug eruption. PMID- 11488717 TI - Allergic contact stomatitis caused by acrylic monomer in a denture. AB - A 71-year-old edentulous man developed a severely painful red mouth at sites of contact with a new denture. Patch testing showed allergy to samples of the denture material and to 2-hydroxyethyl methacrylate. Patch testing to methyl methacrylate was negative. Prolonged boiling of the denture resulted in reversal of his symptoms and samples of this fully cured denture material produced negative patch tests. While allergy to acrylates is a rare cause of stomatitis, this possibility must be considered in patients presenting with oral symptoms. Material safety data sheets are unreliable in providing information regarding the type of acrylate present in the material. Hence, patch testing should be performed with a battery of acrylate allergens as well as with small samples of the denture material. PMID- 11488718 TI - Subcutaneous fat necrosis of the newborn following hypothermia and complicated by pain and hypercalcaemia. AB - A female infant was delivered at term with complications of severe meconium aspiration and birth asphyxia. Surface cooling was performed in the first 24 hours as part of the management of her birth asphyxia. Woody erythema was noted at 24 hours, followed by the formation of red-purple nodules on the 6th day. Clinical findings in the first 24 hours were suggestive of cold panniculitis. However, clinical and histological findings progressed to be in keeping with the diagnosis of subcutaneous fat necrosis of the newborn (SCFN). Furthermore, the immediate postnatal period was complicated by pain resistant to treatment with opiates. Asymptomatic hypercalcaemia was noted on periodic testing at 7 weeks and treated by rehydration, diuretics, prednisolone, etidronate and a low-calcium and -vitamin D diet. A review of the clinical and histological findings of the relevant panniculitides occurring in the postnatal period is presented, as well as a review of the treatment of hypercalcaemia in SCFN. PMID- 11488719 TI - Collagenous and elastotic marginal plaques of the hands. AB - A 49-year-old woman presented with an asymptomatic indurated linear plaque extending from the medial surface of the right thumb along the junction of the dorsal and palmar skin of the hand onto the lateral aspect of the index finger. The left hand showed a similar but less extensive plaque. A skin biopsy showed an acellular zone in the reticular dermis composed of thickened bundles of collagen haphazardly arranged, some perpendicular to the epidermis, admixed with elastic fibres and amorphous basophilic elastotic material. Granular calcium deposits were identified, particularly within degenerate collagen bundles. These clinical and histological features are diagnostic of collagenous and elastotic marginal plaques of the hands, a slowly progressive but largely asymptomatic condition. Actinic degeneration and chronic pressure have been proposed as aetiological agents, but our patient did not exhibit marked actinic degeneration and had no history of chronic occupational pressure. PMID- 11488720 TI - Dowling-Degos disease--a heat aggravated variant. AB - A 22-year-old woman presented with a 5-year history of a micropapular eruption localized to the flexor aspect of her limbs as well as persistent reticulate pigmentation of her neck and upper chest resembling Darier's disease. The eruption was associated with pruritus that was precipitated by heat and was worse in summer. The axillae, groins and inframammary areas had multiple papules but lacked reticulate pigmentation. Multiple biopsies showed an epidermis with club- and antler-like rete ridges but no acantholysis or dyskeratosis. This distinct clinical presentation may represent an unusual heat aggravated variant of Dowling Degos disease that clinically shares features with Darier's disease and transient acantholytic dermatosis. PMID- 11488721 TI - Allergic contact dermatitis due to thiuram exposure from a fungicide. AB - A 49-year-old man developed a widespread eczematous eruption following contact with plants sprayed with a fungicide. Patch testing revealed a strong reaction to thiuram mix and several of the individual thiuram mix constituents. The fungicide contained tetramethylthiuram disulphide, a thiuram chemical. Thiurams are widely recognized as a cause of rubber-glove allergy; however, they are also used extensively in fungicides. PMID- 11488722 TI - Regional lymph node dissection in the treatment of renal cell carcinoma: is it useful in patients with no suspected adenopathy before or during surgery? AB - OBJECTIVES: To evaluate the role of regional lymph node dissection (LND) in a series of patients with renal cell carcinoma (RCC) with no suspicion of nodal metastases before or during surgery. PATIENTS AND METHODS: A series of 167 patients with RCC, free from distant metastases at diagnosis, and who underwent radical nephrectomy at our hospital between January 1990 and October 1997, was reviewed. The mean (median, range) follow-up was 51 (45, 19-112) months. Of the 167 patients, 108 underwent radical nephrectomy alone and 59 had radical nephrectomy with regional LND limited to the anterior, posterior and lateral sides of the ipsilateral great vessel, from the level of the renal pedicle to the inferior mesenteric artery. Of these 59 patients, 49 had no evidence of nodal metastases before or during surgery. The probability of survival was estimated by the Kaplan-Meier method, using the log-rank test to estimate differences among levels of the analysed variables. RESULTS: The overall 5-year survival was 79%; the 5-year survival rate for the 108 patients who underwent radical nephrectomy alone was 79% and for the 49 who underwent LND was 78%. Of the 49 patients with no suspicion of lymph node metastases, one (2%) was found to have histologically confirmed positive nodes. CONCLUSION: These results suggest that there is no clinical benefit in terms of overall outcome in undertaking regional LND in the absence of enlarged nodes detected before or during surgery. PMID- 11488723 TI - A 10-year experience of managing ureteric calculi: changing trends towards endourological intervention--is there a role for open surgery? AB - OBJECTIVES: To study changing practices in the management of ureterolithiasis with the introduction of newer technologies, the efficacy and safety of endourology, extracorporeal shock wave lithotripsy (ESWL) and open surgery, and to determine if any indication remains for open ureterolithotomy in a tertiary endourology unit. PATIENTS AND METHODS: A 12-year retrospective review (1987 1998) was conducted of all primary ureteric stones treated by ESWL, endoscopy, intracorporeal shock wave lithotripsy (ISWL) administered via ureteroscopy, and open surgery. RESULTS: In all, 1195 patients were treated for primary ureteric stones, 44% by ESWL, 37% by ureteroscopy and ISWL, and 20% by open surgery. At the 3-month follow-up the stone-free rates for ESWL monotherapy, ureteroscopy and open surgery were 95%, 85% and 97%, giving an efficiency quotient of 73%, 64% and 94%, respectively. The overall complication rate for ESWL was 13%, for ISWL 32% and for open surgery 13%, but the complications of open surgery were often serious and potentially life-threatening. CONCLUSIONS: With recent advances in endourology the indications for open surgery have decreased considerably, from 26% in 1987-95 to 8% in 1996-98. However, the remaining indications for open ureterolithotomy include failure of less invasive modalities, the presence of medical/anatomical abnormalities, a concomitant open procedure, and the presence of large impacted calculi for which patients prefer to avoid multiple procedures. PMID- 11488724 TI - Acute urinary retention: defining the need and timing for pressure-flow studies. AB - OBJECTIVE: To investigate the utility of "late" pressure-flow studies in predicting the outcome of prostatectomy for acute urinary retention. PATIENTS AND METHODS: Fifty-eight patients with acute urinary retention were prospectively assessed using the International Prostate Symptom Score and pressure-flow studies at a median (range) of 24 (13-60) days after the episode of retention, and before transurethral resection of the prostate. Bladder outlet obstruction and bladder contractility were graded using a modified adaptation of Schafer's passive urethral linear resistance relation. RESULTS: Fifty-six (97%) patients generated a voluntary detrusor contraction, with a mean (range) detrusor pressure at maximum flow of 72.7 (5-144) cmH2O, and 43 (75%) patients were deemed to be obstructed. Eight (16%) patients failed to void after prostatectomy and required clean intermittent catheterization. There were statistically significant differences between successful and unsuccessful patients in mean (SD) age, at 66.30 (6.9) vs 78.8 (2.6) years (P = 0.001), detrusor instability (49% vs 0%, P = 0.01), inability to void during pressure study (8% vs 75%, P = 0.001), and maximal detrusor pressure in the voiding phase, at 80 (36.0) vs 19 (11.2) cmH2O (P = 0.001). CONCLUSIONS: In patients with acute urinary retention, pressure-flow studies undertaken after a period of adequate bladder rest (> 3 weeks) are useful in predicting the surgical outcome. Old age, absence of bladder instability, inability to void during the pressure-flow study and a maximal detrusor pressure of < 20 cmH2O are associated with a poor outcome after prostatectomy. PMID- 11488726 TI - Cimetidine in painful bladder syndrome: a histopathological study. AB - OBJECTIVE: To determine whether oral cimetidine, which reportedly improves symptoms in 60-70% of patients with painful bladder syndrome/interstitial cystitis has a similar mechanism of action on the human bladder and involving a similar peptidergic pathway as it has in human stomach, where it alters histamine gastrin reactions mediated via H2 receptors and a proton pump. PATIENTS AND METHODS: Fourteen patients (13 women and one man, mean age 51 years) with refractory bladder pain and irritative urinary symptoms were treated with cimetidine. The response to cimetidine was assessed by outpatient follow-up and an analogue pain score. Bladder biopsies from eight patients were stained with haematoxylin and eosin, and a polyclonal antibody to gastrin, with counterstaining using toluidine blue, to detect mast cell granules containing histamine. Biopsies from normal areas of the bladder from an age-matched control group of patients with transitional cell carcinoma of the bladder were stained similarly. Human stomach and colon were used as positive controls for gastrin and toluidine blue, respectively. The clinical response to cimetidine was compared with the histology in a blinded fashion. RESULTS: Eight of the patients responded well to cimetidine; none of the biopsies showed evidence of carcinoma in situ. Although G cells in the stomach stained well for gastrin none of the bladder biopsies showed gastrin-like immunoreactivity, apart from some nonspecific urothelial staining. Numerous mast cells with crimson granules and pale nuclei were visible in three patients and fewer in three others; their presence or absence did not correlate with the symptomatic response. CONCLUSIONS: Cimetidine is a useful medical treatment for bladder pain but the presence or absence of gastrin or histamine-like immunoreactivity does not explain its therapeutic benefit. PMID- 11488727 TI - Factors causing variation in urinary N-nitrosamine levels in enterocystoplasties. AB - OBJECTIVE: To establish the presence or absence of any diurnal or long-term variation in N-nitrosamine levels (which might be important in the development of cancer in enterocystoplasties) in enterocystoplasty urine, and to assess other factors that might alter enterocystoplasty N-nitrosamine levels. Patients, subjects and methods Thirty-six patients with enterocystoplasties and six normal controls were assessed. Urine samples were collected every 4 h over a 24-h period and N-nitrosamine levels determined using a modification of the Pignatelli METHOD: An additional urine sample was assessed by microscopy, culture and sensitivity. In a subgroup of 16 patients with an enterocystoplasty, the urinary N-nitrosamine levels were re-measured at 3-monthly intervals. RESULTS: No diurnal or long-term variation in urinary N-nitrosamine levels was identified. The mean N nitrosamine levels were significantly higher in the cystoplasty group than in the controls (1.7 vs 1.0 micromol/L; P = 0.008). Mean N-nitrosamine levels were also significantly higher in enterocystoplasty patients with sterile pyuria than in those with no pyuria (P = 0.01). Those taking prophylactic antibiotics had significantly lower mean N-nitrosamine levels than those not doing so (P = 0.05). Individuals with infected urine and those needing to intermittently catheterize had higher N-nitrosamine levels than their counterparts, but this difference was not significant. Conclusion There were no diurnal or long-term variations in urinary N-nitrosamine levels. Levels were consistently higher in patients with inflamed or infected cystoplasties, those using intermittent self-catheterization and those not taking antibiotic prophylaxis. PMID- 11488728 TI - Does drinking cranberry juice produce urine inhibitory to the development of crystalline, catheter-blocking Proteus mirabilis biofilms? AB - OBJECTIVE: To test the recommendation that to avoid the complications of long term indwelling bladder catheterization (e.g. encrustation and blockage by crystalline Proteus mirabilis biofilms) patients should drink cranberry juice. MATERIALS AND METHODS: Urine was collected from groups of volunteers who had drunk up to 2 x 500 mL of cranberry juice or water within an 8-h period. Laboratory models of the catheterized bladder were supplied with urine from these groups and inoculated with P. mirabilis. After incubation for 24 or 48 h, the extent of catheter encrustation was determined by chemical analysis for calcium and magnesium. Encrustation was also visualized by scanning electron microscopy. RESULTS: The amounts of calcium and magnesium recovered from catheters incubated in urine pooled from individuals who had drunk 500 mL of cranberry juice was not significantly different from that on catheters incubated in pooled urine from control subjects who had drunk 500 mL of water. However, there was significantly less encrustation (P = 0.007) on catheters from models receiving urine from volunteers who had drunk 2 x 500 mL of water than on catheters incubated in models supplied with urine from volunteers who had drunk 2 x 500 mL of cranberry juice. The amounts of encrustation on these two groups of catheters were also significantly less than that on catheters incubated in models supplied with urine from volunteers who had not supplemented their normal fluid intake. (P < 0.001). Experiments in the models using artificial urine showed that increasing the low fluid intake (720 mL/24 h) characteristic of many patients undergoing long-term catheterization by factors of three and six, significantly (P < 0.01) reduced the amounts of calcium and magnesium that formed on catheters. At a simulated fluid intake of 720 mL/24 h, catheters blocked with encrustation after a mean of 42.5 h, while those supplied with urine produced from an intake of 4320 mL/24 h, drained freely for > 10 days. CONCLUSION: In this in vitro study, drinking cranberry juice did not produce urine that was inhibitory to the development of crystalline catheter-blocking P. mirabilis biofilms. The important factor in preventing catheter encrustation is a high fluid intake. PMID- 11488729 TI - Human papillomaviruses 6/11, 16/18 and 31/33/51 are not associated with squamous cell carcinoma of the urinary bladder. AB - OBJECTIVE: To assess high-risk human papillomavirus (HPV), mainly HPV type 16, 18, 31 and 33 (an important aetiological factor in squamous cell carcinoma, SCC, of the anogenital region) in SCC of the urinary bladder. MATERIAL AND METHODS: Sixteen SCC from the urinary bladder were evaluated using non-isotopic in situ hybridization with a sensitive detection system for the presence of high-risk HPV 16/18, or 31/33/51, and for HPV6/11, a low-risk type commonly found in condylomata. Previously published studies were also reviewed and assessed. RESULTS: No high-risk HPV was found in any of the SCC of the bladder evaluated. Previous reports identified nine HPV-positive SCC of a total of 105, including the present series. In four of these positive cases, HPV types were found that are considered a high risk in anogenital carcinomas. CONCLUSION: From the present and previous results, we conclude that HPV has no major role in the pathogenesis of SCC of the urinary bladder. PMID- 11488730 TI - p53, p21 and mdm2 expression vs the response to radiotherapy in transitional cell carcinoma of the bladder. AB - OBJECTIVE: To identify, in a retrospective study, possible molecular markers predictive of radioresponsiveness in patients with transitional cell carcinoma (TCC) of the bladder. PATIENTS AND METHODS: Patients with T2-T4a TCC treated with preoperative radiotherapy and cystectomy were included in the study if their cystectomy specimen was pT3b (in 42) or pT0 (in 17). Because treatment schedules changed over time, radiotherapy was given either as 2 Gy x 23 over 4-5 weeks with cystectomy 4-5 weeks later (in 23), or as 4 Gy x 5 during 1 week with cystectomy in the following week (in 36 patients). Protein expression of p53, mdm2 and p21 (CDKN1 A/WAF1/CIP1/SDI1) was assessed immunohistochemically in biopsies taken before radiotherapy. RESULTS: There was no difference in protein expression when comparing all patients with pT0 and pT3b. However, for patients receiving 46 Gy, increased p53 expression (but not p21 or mdm2) predicted the absence of residual tumour (P = 0.005): six of seven patients with > 50% p53 expression had pT0 in the cystectomy specimen, whereas 10 of 12 patients with < or = 5% expression had pT3b. Over-expression of p53 correlated with longer overall (P = 0.045) and cancer-specific survival (P = 0.020). CONCLUSION: The expression of mdm2 or p21 did not predict radioresponsiveness in patients with TCC of the bladder. The role of p53 remains unclear; the view that p53 over-expression confers radioresistance in bladder cancer is not supported. PMID- 11488731 TI - A systematic review of intravesical bacillus Calmette-Guerin plus transurethral resection vs transurethral resection alone in Ta and T1 bladder cancer. AB - OBJECTIVE: To assess, in a systematic review, the effectiveness of intravesical bacillus Calmette-Guerin (BCG) in preventing tumour recurrence in patients with medium/high risk Ta and T1 bladder cancer. PATIENTS AND METHODS: An electronic database search of Medline, Embase, DARE, the Cochrane Library, Cancerlit, Healthstar and BIDS was undertaken, plus hand searching of the Proceedings of ASCO, for randomized controlled trials, in any language, comparing transurethral resection (TUR) alone with TUR followed by intravesical BCG in patients with Ta and T1 bladder cancer. RESULTS: The search identified 26 publications comparing TUR with TUR + BCG. Six trials were considered acceptable, representing 585 eligible patients, 281 in the TUR-alone group and 304 in the TUR + BCG group. The major clinical outcome chosen was tumour recurrence. The weighted mean log hazard ratio for the first recurrence, taken across all six trials, was -0.83 (95% confidence interval -0.57 to -1.08, P < 0.001), which is equivalent to a 56% reduction in the hazard, attributable to BCG. The Peto odds ratio for patients recurring at 12 months was 0.3 (95% confidence interval of 0.21-0.43, P < 0.001), significantly favouring BCG therapy. Manageable toxicities associated with intravesical BCG were cystitis (67%), haematuria (23%), fever (25%) and urinary frequency (71%). No BCG-induced deaths were reported. CONCLUSION: TUR with intravesical BCG provides a significantly better prophylaxis of tumour recurrence in Ta and T1 bladder cancer than TUR alone. Randomized trials are still needed to address the issues of BCG strain, dose and schedule, and to better quantify the effect on progression to invasive disease. PMID- 11488732 TI - The 2-year symptomatic and urodynamic results of a prospective randomized trial of interstitial radiofrequency therapy vs transurethral resection of the prostate. AB - OBJECTIVE: To assess the 2-year symptomatic and urodynamic results of a prospective randomized trial of interstitial radiofrequency therapy of the prostate (IRFT) vs transurethral resection of the prostate (TURP). PATIENTS AND METHODS: Patients with lower urinary tract symptoms (LUTS) and urodynamic evidence of bladder outlet obstruction (BOO) were randomized to undergo IRFT or TURP and were followed up using the International Prostate Symptom Score (IPSS) and urodynamic assessment for 2 years. RESULTS: At 2 years there was a clinically relevant reduction in the IPSS in the IRFT (20 to 9) and TURP groups (22 to 4). There was also a statistically significant reduction in the detrusor pressure at maximum urinary flow in both groups, but the reduction in the IRFT group was probably not sufficient to explain the observed symptomatic improvements solely from a reduction in BOO. CONCLUSION: IRFT can produce a sustained improvement in LUTS for at least 2 years. However, such improvements are unlikely to be entirely the result of a reduction in BOO. The effects of radiofrequency energy may, at least partly, be independent of any thermal effect and depend instead on neuromodulation of lower urinary tract nerves. PMID- 11488733 TI - The percentage of free prostate-specific antigen does not predict extracapsular disease in patients with clinically localized prostate cancer before radical prostatectomy. AB - OBJECTIVE: To determine whether the percentage of free/total prostate-specific antigen (f/tPSA) can predict the pathological features in patients with clinically localized prostate cancer before radical prostatectomy. PATIENTS AND METHODS: Univariate and multivariate logistic regression was used to analyse data from 171 untreated patients who underwent radical prostatectomy. Variables included the total PSA (tPSA), fPSA, f/tPSA, biopsy Gleason score, clinical stage and patient age. RESULTS: In 115 patients with pathologically organ-confined tumours ( pT2N0) the mean (SD) tPSA value was 6.9 (5.6) ng/mL; in 56 patients with extracapsular disease ( pT3pN0/N+) it was 10.2 (7.6) ng/mL; the respective f/tPSA values were 14.9 (8.1)% and 14.2 (12.9)%. In the univariate and multivariate analysis, tPSA and biopsy Gleason score were highly significant in predicting extracapsular disease (P < 0.001 and 0.002) but the f/tPSA was not (P = 0.18). There was no significant difference between the mean f/tPSA and final Gleason scores. CONCLUSION: The f/tPSA does not predict extracapsular disease in patients with clinically localized prostate cancer before radical prostatectomy. Knowing the f/tPSA provides no significant additional information in predicting extracapsular disease when the biopsy Gleason score and tPSA are known. PMID- 11488734 TI - Routine bone scans in patients with prostate cancer related to serum prostate specific antigen and alkaline phosphatase. AB - OBJECTIVE: To evaluate the need for a bone scan as a routine staging procedure in patients with newly diagnosed prostate cancer in relation to serum prostate specific antigen (PSA) and alkaline phosphatase (ALP) levels, and thus determine whether a reduction of the use of this staging method is possible in patients with a low probability of osseous metastasis. PATIENTS AND METHODS: The results of bone scans were related retrospectively to levels of serum PSA and ALP in 363 patients with prostate cancer newly diagnosed between 1989 and 1997. RESULTS: Of 363 consecutive patients, 111 had a positive bone scan. In 19 of 144 (13%, "missed diagnosis") patients with a PSA level of < 20 ng/mL the bone scan was positive. In 125 patients (49%, "false-positives") with a PSA level of > 20 ng/mL the bone scan was negative. A threshold level of 100 U/L for ALP gave a better balance for the number of "false-positives" and "missed diagnosis". ALP values correlated better with an abnormal bone scan than did PSA levels; ALP levels of > 90 U/L indicated a 60% chance for the presence of bone metastases. CONCLUSION: Patients with newly diagnosed and untreated prostate cancer should undergo bone scintigraphy if there is bone pain or if ALP levels are > 90 U/L. Recent reports discourage the routine use of a bone scan when the serum PSA level is <20 ng/mL. However, the present series suggests there is a greater chance of a positive bone scan in patients with low PSA levels; these findings need further confirmation. PMID- 11488735 TI - Pretreatment levels of urinary deoxypyridinoline as a potential marker in patients with prostate cancer with or without bone metastasis. AB - OBJECTIVE: To assess the predictive role of the bone markers alkaline phosphatase (ALP) and urinary deoxypyridinoline (DPD), as indicators of bone turnover, at baseline in patients with prostate cancer. PATIENTS, SUBJECTS AND METHODS: Urinary DPD, serum ALP and prostate-specific antigen (PSA) were evaluated in 23 patients with benign prostatic hyperplasia (BPH), 115 with prostatic carcinoma, of whom 21 had bone metastasis, and in 16 age-matched control subjects. RESULTS: Patients with newly diagnosed prostate cancer and bone metastasis had a higher urinary excretion of DPD, and a higher serum PSA and ALP than had patients with BPH and those with prostate cancer but no metastasis. Receiver operating curve analysis for PSA, ALP and DPD showed a significant discriminating ability for positive and negative bone scans (P = 0.0684). However, from logistic regression of the combinations, only serum ALP was a significant independent predictor of bone metastasis in patients with prostate cancer. CONCLUSION: Serum ALP or urinary DPD are the best predictors of bone metastasis in patients with prostate cancer; further studies with more patients are required. PMID- 11488736 TI - Modified plication of the tunica albuginea in treating congenital penile curvature. AB - OBJECTIVE: To describe a modified plication technique and compare the results with the Nesbit procedure for treating congenital penile curvature. PATIENTS AND METHODS: Eleven men with congenital penile curvature were treated by a modified tunica albuginea plication technique. An artificial erection was induced by normal saline injection; in the areas where plication was planned, Buck's fascia was opened longitudinally and dissected a short distance toward the midline to free the neurovascular bundles. Allis clamps were used to grasp the tunica albuginea and "bumps" created. Two interrupted U-shaped sutures using 2/0 polyglactin were placed underneath each Allis clamp to secure the bump. The results were analysed retrospectively and compared with the results of 11 other men treated using the Nesbit procedure. RESULTS: The mean (range) follow-up was 25 (12-47) months for the Nesbit procedure and 15 (8-26) months for the modified plication procedure. In the Nesbit group, eight patients had satisfactory cosmetic and functional results; three complained of penile shortening and one had erectile dysfunction. In the modified plication group, 10 patients reported satisfactory cosmetic and functional results; one complained of penile shortening, two were concerned about the indurations of the penis but none had erectile dysfunction. CONCLUSIONS: The modified plication technique is easier to perform, is more often successful and causes fewer surgical complications than the Nesbit procedure for treating congenital penile curvature. PMID- 11488737 TI - Treatment with sildenafil citrate in renal transplant patients with erectile dysfunction. AB - OBJECTIVE: To assess the safety and efficacy of sildenafil citrate in renal transplant patients with erectile dysfunction, as up to half of men with renal failure may be affected and only 60-75% recover potency after transplantation. PATIENTS AND METHODS: Fifty patients with erectile dysfunction and a functioning renal transplant were treated using sildenafil (mean age 54 years, mean time on dialysis 35 months, mean time from transplantation 20 months). The hypogastric artery was not used during transplantation in any patient. Sildenafil citrate was prescribed at doses of 25 or 50 mg depending on baseline creatinine values and on the response, and plasma levels of cyclosporin/FK506 were monitored. RESULTS: Thirty patients (60%) had a satisfactory response, with a mean time on dialysis of 23 months. Six patients (12%) did not take the sildenafil and in 14 (28%) the drug was ineffective. The mean time on dialysis in this group was 43 months. Six patients (12%) had side-effects that in no case led to withdrawal of treatment. Plasma levels of cyclosporin/FK506 remained within the safety and efficacy limits in all patients. CONCLUSIONS: Treatment with sildenafil citrate in renal transplant patients with erectile dysfunction is an effective and safe option, with few side-effects. Plasma levels of immunosuppressants are unchanged. The response was more effective in patients with a shorter time on dialysis, as penile vascular disease is less advanced. PMID- 11488738 TI - Effect of radical retropubic prostatectomy on erectile function, evaluated before and after surgery using colour Doppler ultrasonography and nocturnal penile tumescence monitoring. AB - OBJECTIVE: To assess the effect of radical retropubic prostatectomy on erectile function, by evaluating objectively patients' erectile function before and after surgery. PATIENTS AND METHODS: The study comprised 126 patients with clinically localized prostate cancer who were scheduled to undergo radical retropubic prostatectomy. After giving informed consent for the study, 123 patients underwent intracavernosal injection tests, colour Doppler ultrasonography and nocturnal penile tumescence monitoring before and after surgery. RESULTS: From the intracavernosal injection tests and nocturnal penile tumescence monitoring, 21 patients (17%) were evaluated as having normal erectile function before surgery. After radical retropubic prostatectomy, nine (43%) of these 21 potent men had preserved erectile function. In eight patients whose neurovascular bundles were preserved, five were potent after surgery. The cause of erectile function after surgery was a neurogenic disorder in seven and a related vascular disorder in five. CONCLUSION: From objective tests of erectile function on patients scheduled to undergo radical prostatectomy, 17% had normal erectile function. However, even after nerve-sparing radical retropubic prostatectomy, the proportion retaining potency was unsatisfactory. Although a neurological disorder was the main cause of erectile dysfunction after surgery, vascular disorders were also important. PMID- 11488739 TI - Simple intratesticular cysts in adults: a diagnostic dilemma. PMID- 11488740 TI - Benign scrotal pathology: should all patients undergo surgery? AB - OBJECTIVE: To evaluate the justification for operative intervention in patients undergoing surgery for benign scrotal pathology, and to assess the associated morbidity. PATIENTS AND METHODS: Consecutive patients in two surgical firms who underwent surgery for benign intrascrotal pathology over a 5-year period were studied retrospectively. RESULTS: In all, 102 patients were included in the study (mean age 51.6 years, range 16-86). Most patients had either epididymal cysts (59) or hydroceles (31) or a combination of the two (eight). Some patients (37) underwent ultrasonography before surgery. The indications for surgery were deemed to be strong in 25% of patients but weak or absent in the remainder. Complications occurred in 31 patients, being minor in 18, but significant in 13, resulting in four re-admissions to hospital. There was no significant difference in the complication rate between those patients with strong indications for surgery (37%) and those with weak indications (28%). CONCLUSIONS: Surgery for benign intrascrotal pathology is frequently undertaken for weak clinical indications and carries significant associated morbidity. This could be avoided in many cases by simple reassurance. Judicious use of ultrasonography is advised. A policy of selective surgical intervention is strongly advocated. PMID- 11488741 TI - HOSE: an objective scoring system for evaluating the results of hypospadias surgery. AB - OBJECTIVE: To determine the accuracy and utility of a scoring system designed to allow an objective appraisal of the outcome of hypospadias repair, based on evaluating meatal location, meatal shape, urinary stream, straightness of erection, and the presence and complexity of any complicating urethral fistula. PATIENTS AND METHODS: Twenty patients (median age 23 months) were randomly selected and reviewed at a median of 8.9 months after their hypospadias repair. Two paediatric surgeons, a nurse and one of the child's parents independently assessed each patient using the "hypospadias objective scoring evaluation" (HOSE) system. The results were collated and the level of interobserver variation assessed using the weighted kappa test. RESULTS: The mean weighted kappa was 0.66, indicating good agreement among observers. The level of agreement was highest between surgeon and nurse at 0.70, but remained good between surgeon and parent, at 0.65. CONCLUSIONS: Interobserver variation using the HOSE system was minimal, supporting its use as an objective outcome measure after hypospadias surgery, and facilitating an impartial evaluation of operations used in correcting hypospadias. PMID- 11488743 TI - The clinical and radiological findings in boys with endoscopically severe congenital posterior urethral obstruction. AB - OBJECTIVE: To assess the clinical and radiological spectrum in boys with endoscopically severe posterior urethral obstruction, as there is a broad clinical spectrum which does not always correlate with the cystoscopic findings. PATIENTS AND METHODS: Between December 1990 and July 2000, 39 boys (newborn to 12 years old) underwent cystoscopy to investigate a urethral anomaly and were found to have a severe obstructing posterior urethral membrane. Their voiding cystograms, video-recorded cystoscopy and presenting signs were reviewed. RESULTS: Of the 39 boys assessed, a voiding cystogram and presenting signs were available in 36. Of these 36 boys, three cystograms were initially reported as normal, eight showed posterior urethral dilatation as the only feature, and the remainder had more severe bladder and upper tract changes. Fourteen were diagnosed after identifying antenatal hydronephrosis, four presented with voiding dysfunction, one with haematuria and 17 were found after investigation of a urinary tract infection. CONCLUSIONS: This study shows that a congenital posterior urethral membrane that has only a small posterior defect endoscopically can have a wide clinical and radiological spectrum. The difference in outcome may be caused by variations in the response of the bladder to the obstruction, resulting in a different effect on the upper tracts. PMID- 11488742 TI - Mercurochrome as an adjunct to local preoperative preparation in children undergoing hypospadias repair. AB - OBJECTIVE: To determine whether the local application of mercurochrome over genital skin before surgery, in addition to soap/water scrubs, can help to decrease the incidence of infection patients undergoing hypospadias repair, and thereby the incidence of fistula formation. PATIENTS AND METHODS: The study comprised 44 consecutive boys undergoing primary or repeat surgery for hypospadias between October 1999 and April 2000. They were arbitrarily divided into two groups; group 1 received a conventional local scrub with soap/water 48 h before surgery and group 2, a conventional local scrub with soap/water was followed by a local application of 2% mercurochrome for 48 h before surgery. Both groups were comparable in age, location of the meatus and stage of repair. All patients were exposed to similar conditions during and after surgery. Urine, skin and wound swabs taken before during and after surgery were assessed microbiologically, using standard precautions. The incidence of infection and complications was then compared. RESULTS: Mercurochrome significantly decreased local infection, especially before surgery, but its effect in decreasing fistula formation, although appreciable, was not statistically significant. CONCLUSION: The local application of mercurochrome after a soap/water scrub for at least 48 h before surgery is a simple, economic and effective means to decrease postoperative wound infection. Future studies are needed, keeping other determinants of fistula formation constant, to evaluate its role in decreasing fistula formation in patients undergoing hypospadias repair. PMID- 11488744 TI - An autoradiographic study of regional blood flow distribution in the rat kidney during ureteric obstruction--the role of vasoactive compounds. AB - OBJECTIVE: To determine the changes in regional renal blood flow during ureteric obstruction and to examine the role of vasoactive mediators in effecting these changes. MATERIALS AND METHODS: Renal blood flow in Sprague-Dawley rats was assessed after periods of ureteric obstruction using a quantitative autoradiographic technique based on Kety's theory of diffusion of an inert tracer (14C-iodoantipyrine). Prostaglandins, thromboxanes and renin-angiotensin were inhibited pharmacologically using diclofenac sodium and enalapril. RESULTS: Baseline blood flow to the outer cortex, inner cortex and medulla was 807, 258 and 105 mL/100 g/min, respectively. There was an increase in outer cortical blood flow after 10 min of ureteric obstruction which became significant at 30 min (P < 0.05). There was a significant decrease in inner cortical and medullary blood flow at 30 min, to 210 and 68 mL/100 g/min, respectively (P < 0.05). Diclofenac sodium abolished the increase in outer cortical blood flow. After 24 h of unilateral ureteric obstruction, outer cortical blood flow decreased to 492 mL/100 g/min; inner cortical blood flow also decreased but to a lesser extent, to 190 mL/100 g/min. Inhibition of prostaglandins, thromboxanes and the renin angiotensin system reduced the degree of renal vasoconstriction but there was still a significant decrease in outer cortical perfusion despite the presence of these blocking agents. CONCLUSIONS: The control of the renal vasculature involves a complex interplay between a variety of vasoactive mediators. Quantitative autoradiography offers the opportunity to evaluate changes in regional renal perfusion with high resolution and will allow a greater understanding of the pathophysiology of renal diseases. PMID- 11488745 TI - Effects of L-arginine on the kidney levels of malondialdehyde in rats submitted to renal ischaemia-reperfusion. AB - OBJECTIVE: To evaluate the effects of L-arginine, a nitric oxide donor, on kidney levels of malondialdehyde (MDA, a product of cellular lipid peroxidation), serum creatinine levels, and urinary volume in rats undergoing unilateral renal ischaemia-reperfusion. MATERIALS AND METHODS: Wistar rats (117) were randomly distributed into three experimental groups (of four subgroups each) in which were assessed renal cell-lipid peroxidation (kidney levels of MDA), serum creatinine levels and urinary volume. The rats underwent unilateral nephrectomy followed by contralateral renal ischaemia-reperfusion with or with no pretreatment with L arginine (200 mg/kg) given intraperitoneally. RESULTS: Pretreatment with L arginine caused significantly higher kidney levels of MDA than in the untreated group (P < 0.05). Furthermore, L-arginine given before surgery attenuated the increase in serum creatinine and significantly increased urinary volume in rats subjected to renal ischaemia-reperfusion (P < 0.05). CONCLUSION: L-arginine tended to be of benefit for renal function during renal ischaemia-reperfusion in rats. Pretreatment with L-arginine (200 mg/kg intraperitoneally) seems to increase the renal damage by increasing kidney levels of MDA. PMID- 11488747 TI - Minimally invasive harvesting of fascia lata for use in the pubovaginal sling procedure. PMID- 11488746 TI - Chronic ethanol consumption impairs adrenoceptor- and purinoceptor-mediated relaxations of isolated rat detrusor smooth muscle. AB - OBJECTIVE: To investigate the effects of chronic ethanol consumption on the reactivity of detrusor smooth muscle. MATERIALS AND METHODS: Eight male rats received ethanol (7.2% v/v) in a modified liquid diet for 4 weeks. Two control groups were assessed; eight rats in one group were fed sucrose and received a liquid diet, and 12 rats in the second group received standard rat chow and water for 4 weeks. The reactivity of detrusor smooth muscle strips from ethanol-fed animals and control animals was evaluated in organ chambers. RESULTS: The relaxation response elicited by isoprenaline or adenosine was unaffected in the both control groups while it was significantly inhibited, with decreased maximum responses and pD2 values, in the ethanol-fed group. Contractile responses of detrusor smooth muscle to carbachol or 80 mmol/L KCl and relaxant responses to papaverine were similar in the control groups and the ethanol-fed group. There was no change in agonist potency among the groups. CONCLUSION: Chronic ethanol consumption impairs beta-adrenoceptor- and purinoceptor-mediated relaxation but not cholinoceptor-mediated contraction of the rat detrusor smooth muscle. Thus, it appears that different regulatory mechanisms are involved in ethanol-induced alterations in beta-adrenergic, purinergic and muscarinic receptors in detrusor strip. PMID- 11488748 TI - Oxidized cellulose haemostat mimicking a possible recurrence of neuroblastoma. PMID- 11488749 TI - Failure of carbon dioxide laser treatment in three patients with penoscrotal extramammary Paget's disease. PMID- 11488750 TI - Robotics in urology: the Ethicon Foundation Fund Travelling Fellowship, the Royal College of Surgeons of Edinburgh. PMID- 11488751 TI - Medical linguist. PMID- 11488752 TI - The management of intractable haematuria. PMID- 11488754 TI - A prospective evaluation of the pathogenesis of detrusor instability in women, using electron microscopy and immunohistochemistry. PMID- 11488753 TI - The management of intractable haematuria. PMID- 11488756 TI - The effect of superoxide dismutase on nitric oxide-mediated and electrical field stimulated diabetic rabbit cavernosal smooth muscle relaxation. PMID- 11488759 TI - Reduction of paraphimosis the simple way--the Dundee technique. PMID- 11488761 TI - Relaxation induced by omeprazole does not change in diabetic rabbit corpus cavernosum. PMID- 11488762 TI - Prostatic small-cell neuroendocrine carcinoma with disease progression monitored by measurement of serum progastrin-releasing peptide. PMID- 11488763 TI - Benign prostatic hyperplasia is a reawakened process of persistent Mullerian duct mesenchyme. PMID- 11488765 TI - Urethral metastasis after transurethral resection of a malignant prostate. PMID- 11488767 TI - The use of sputum cell counts to evaluate asthma medications. AB - Total and differential cell counts from hypertonic-induced, dithiothreitol dispersed sputum provide reproducible measurements of airway inflammatory cell counts, which are responsive to treatment with anti-inflammatory drugs. They have helped to understand the kinetics of inflammatory cell changes in asthma after the reduction of corticosteroids and the subsequent re-introduction of treatment. They have identified that the presence of sputum eosinophilia in asthma, chronic cough and chronic airflow limitation is a predictor of steroid-responsiveness and of lack of 'asthma control'. They can be used to study the dose-response effect of inhaled corticosteroids and may be useful to establish the relative potency of different corticosteroid formulations and delivery devices. Sputum cell counts are also useful to study the potential anti-inflammatory effects of drugs like theophylline, long-acting beta-adrenoceptor agonists, leukotriene antagonists and newer drugs in development. They may be helpful to select add-on therapy to corticosteroids in 'difficult-to-control' asthma. PMID- 11488768 TI - Effects of anticancer drugs on the metabolism of the anticancer drug 5,6 dimethylxanthenone-4-acetic (DMXAA) by human liver microsomes. AB - AIMS: To investigate the effects of various anticancer drugs on the major metabolic pathways (glucuronidation and 6-methylhydroxylation) of DMXAA in human liver microsomes. METHODS: The effects of various anticancer drugs at 100 and 500 microM on the formation of DMXAA acyl glucuronide (DMXAA-G) and 6-hydroxymethyl-5 methylxanthenone-4-acetic acid (6-OH-MXAA) in human liver microsomes were determined by high performance liquid chromatography (h.p.l.c.). For those anticancer drugs showing significant inhibition of DMXAA metabolism, the inhibition constants (Ki) were determined. The resulting in vitro data were extrapolated to predict in vivo changes in DMXAA pharmacokinetics. RESULTS: Vinblastine, vincristine and amsacrine at 500 microM significantly (P < 0.05) inhibited DMXAA glucuronidation (Ki = 319, 350 and 230 microM, respectively), but not 6-methylhydroxylation in human liver microsomes. Daunorubicin and N-[2 (dimethylamino)-ethyl]acridine-4-carboxamide (DACA) at 100 and 500 microM showed significant (P < 0.05) inhibition of DMXAA 6-methylhydroxylation (Ki = 131 and 0.59 microM, respectively), but not glucuronidation. Other drugs such as 5 fluoroucacil, paclitaxel, tirapazamine and methotrexate exhibited little or negligible inhibition of the metabolism of DMXAA. Pre-incubation of microsomes with the anticancer drugs (100 and 500 microM) did not enhance their inhibitory effects on DMXAA metabolism. Prediction of DMXAA-drug interactions in vivo based on these in vitro data indicated that all the anticancer drugs investigated except DACA appear unlikely to alter the pharmacokinetics of DMXAA, whereas DACA may increase the plasma AUC of DMXAA by 6%. CONCLUSIONS: These results indicate that alteration of the pharmacokinetics of DMXAA appears unlikely when used in combination with other common anticancer drugs. However, this does not rule out the possibility of pharmacokinetic interactions with other drugs used concurrently with this combination of anticancer drugs. PMID- 11488769 TI - The relationship between metformin therapy and the fasting plasma lactate in type 2 diabetes: The Fremantle Diabetes Study. AB - AIMS: To determine (i) which factors, including metformin, are associated with the fasting plasma lactate concentration in type 2 diabetes, and (ii) whether plasma lactate is associated with haemodynamic and metabolic effects. METHODS: We measured fasting plasma lactate in 272 well-characterized diabetic patients from a community-based sample, 181 (67%) of whom were taking metformin with or without other therapies. Linear regression analysis was used to identify predictors, including metformin therapy, of the plasma lactate, and to investigate associations between plasma lactate and resting pulse rate and serum bicarbonate. Factor analysis assessed independent relationships between groups of cosegregating variables. RESULTS: Metformin-treated patients had higher plasma lactate concentrations than nonmetformin-treated subjects (geometric mean [s.d. range] 1.86 [1.34-2.59] vs 1.58 [1.09-2.30] mmol x l(-1), respectively; P < 0.001). In a linear regression model, plasma glucose, BMI and metformin use (but not dose) were independently associated with plasma lactate (P < or = 0.028); after adjustment for the former two variables, metformin-treated patients had a mean plasma lactate 0.16 mmol l-1 greater than in subjects not taking the drug. Factor analysis revealed that plasma lactate, plasma glucose, BMI and pulse rate cosegregated but serum bicarbonate was not in this grouping. CONCLUSIONS: The present results show that metformin therapy increases the fasting plasma lactate in ambulant patients with type 2 diabetes from a community-based cohort. From associations in the data we hypothesize that this increase reflects (i) increased sympathetic activity in patients with the metabolic syndrome (ii) increased substrate (glucose) availability and (iii) a direct metformin effect. PMID- 11488770 TI - Cigarette smoking in men and vascular responsiveness. AB - AIMS: Smoking is a major risk factor for developing atherosclerosis. In order to understand the vascular abnormalities observed in smokers, we investigated vascular responsiveness in cigarette smokers. METHODS: We performed two consecutive matched group comparative studies to investigate vascular responsiveness using venous occlusion plethysmography. The mean effects of three incremental doses of each vasoactive agent are presented. Both studies compared smokers with nonsmokers. RESULTS: The first investigated 68 subjects (smokers = 29; mean +/- s.d. ages; 24 +/- 6 vs 25 +/- 5 years; P = NS) and found smoking was associated with a significant blunting of the flow ratio between treated and untreated arms to endothelium-dependent vasodilatation to acetylcholine (mean +/- s.d., nonsmokers vs smokers) 4.07 +/- 2.18 vs 3.42 +/- 1.79 (P = 0.04, 95% CI 0.02, 1.12). By contrast, there was no significant difference in the responses to the endothelium-independent vasodilators sodium nitroprusside and verapamil. Smoking was also associated with a significant impairment in endothelium dependent vasoconstriction induced by monomethyl-L-arginine (L-NMMA) 0.78 +/- 0.22 vs 0.87 +/- 0.21 (P = 0.006, 95% CI -0.14, -0.02) and a trend to blunted endothelium-independent vasoconstrictor responses to noradrenaline. In the second study we investigated the response to angiotensin I and II in 23 subjects (smokers = 12; mean +/- s.d. ages; 34 +/- 10 vs 32 +/- 11 years). There was significant impairment in smokers of the mean vasoconstrictor response to angiotensin I 0.51 +/- 0.15 vs 0.59 +/- 0.16 (nonsmokers vs smokers; P = 0.003, 95% CI -0.13, -0.03) and a nonsignificant trend towards impairment of the response to angiotensin II. CONCLUSIONS: Cigarette smoking in male volunteers is associated with blunted basal and stimulated nitric oxide bioactivity. Endothelial independent vasodilator responses (to nitroprusside and verapamil) were unaltered in smokers. A defect in the vasoconstrictor response to angiotensin I was also seen. PMID- 11488771 TI - Endothelin-A receptor antagonist inhibits angiotensin II and noradrenaline in man. AB - AIMS: Endothelin-1 (ET-1) is a potent vasoconstrictor produced by the vascular endothelium. The interactions of ET with the mediators of the sympathetic nervous system and the renin-angiotensin-system in humans are unclear. METHODS: We studied the effects of the ETA-selective antagonist BQ-123 and the ETB-selective antagonist BQ-788 (both 10(-10)-10(-8) M) on ET-1 (10(-16)-10(-10) M), angiotensin II (AT, 10(-16)-10(-10) M) and noradrenaline (NA, 10(-16)-10(-10) M) induced vasoconstriction in the human skin microcirculation in vivo in 25 healthy male volunteers using laser Doppler flowmetry and double injection technique. RESULTS: BQ-123 caused a dose-dependent vasodilatation (maximum effect: + 949 +/- 84 AUC-PU, P < 0.001), whereas BQ-788 induced mild vasoconstriction (maximum effect: -388 +/- 96 AUC-PU, P < 0.01). In the presence of BQ-123, but not BQ-788, ET-1, AT and NA caused markedly less vasoconstriction at any tested agonist dose; the effect was most pronounced on ET-1 (maximum effect at 10(-14) M: + 814 +/- 93 AUC-PU vs ET alone, P < 0.001), followed by noradrenaline (maximum effect at 10( 16) M: +580 +/- 107 AUC-PU vs NA alone, P < 0.01) and angiotensin II (maximum effect at 10(-14) M: + 493 +/- 111 AUC-PU vs AT alone, P < 0.001). CONCLUSIONS: ETA-selective antagonism inhibits vasoconstriction to AT and NA in vivo in healthy subjects. This beneficial effect may be useful for the treatment of patients with cardiovascular disease including hypertension especially in combination therapy with sympatholytic agents and inhibitors of the renin angiotensin system. PMID- 11488772 TI - Adrenomedullin (ADM) in the human forearm vascular bed: effect of neutral endopeptidase inhibition and comparison with proadrenomedullin NH2-terminal 20 peptide (PAMP). AB - AIMS: To compare the haemodynamic responses of proadrenomedullin N-terminal 20 peptide (PAMP) and adrenomedullin (ADM) in the forearm vascular bed of healthy male volunteers, and to investigate the role of neutral endopeptidase (NEP) in the metabolism of ADM. METHODS: On two separate occasions, ADM (1-30 pmol x min( 1)) and PAMP (100-3000 pmol x min(-1)) were infused into the brachial artery of eight male subjects, and forearm blood flow (FBF) assessed using venous occlusion plethysmography. In a second study, eight male subjects received the same doses of ADM, co-infused with either the NEP inhibitor thiorphan (30 nmol x min(-1)) or the control vasoconstrictor noradrenaline (120 pmol x min(-1)), on separate occasions. Both studies were conducted in a double-blind, randomized manner. RESULTS: ADM and PAMP produced a dose-dependent increase in FBF (P < or = 0.002). Based on the dose producing a 50% increase in FBF, ADM was approximately 60 times more potent than PAMP. Thiorphan and noradrenaline produced similar reductions in FBF of 14 +/- 4% (mean +/- s.e. mean) and 22 +/- 6%, respectively (P = 0.4). However, the area under the dose-response curve was significantly greater during co-infusion of ADM with thiorphan than with noradrenaline (P = 0.028), as was the maximum increase in FBF ratio (2.1 +/- 1.0 vs 1.2 +/- 0.2; P = 0.030). CONCLUSIONS: ADM and PAMP both produce a local dose-related vasodilatation in the human forearm, but PAMP is approximately 60 times less potent than ADM. In addition, NEP inhibition potentiates the haemodynamic effects of ADM. These findings suggest that PAMP may not play a role in the physiological regulation of blood flow. However, in pathophysiological conditions such as hypertension and heart failure, NEP inhibition may exert a beneficial effect by increasing the biological activity of ADM. PMID- 11488773 TI - Effects of adrenomedullin on angiotensin II stimulated atrial natriuretic peptide and arginine vasopressin secretion in healthy humans. AB - AIMS: Adrenomedullin is a newly described peptide that has widespread tissue distribution. Its presence in cardiovascular (including vascular endothelial cells, smooth muscle cells, and cardiac atria and ventricles) and renal tissues, together with its vasodilatory and natriuretic properties, suggest a role in blood pressure regulation and fluid and electrolyte balance. METHODS: Nine normal volunteers were studied to determine whether or not adrenomedullin influenced plasma atrial natriuretic peptide and arginine vasopressin concentrations during systemic angiotensin II infusion. RESULTS: A significant (P = 0.02) augmentation of atrial natriuretic peptide concentrations, but no suppression of arginine vasopressin concentrations, was found with coinfusion of adrenomedullin and angiotensin II when compared with vehicle and angiotensin II. CONCLUSIONS: Despite its vasodilator and natriuretic action, adrenomedullin significantly augmented angiotensin II-stimulated plasma atrial natriuretic peptide concentrations in healthy humans. This provides further evidence of a synergistic interaction between adrenomedullin and atrial natriuretic peptide and suggests that adrenomedullin may have a role in fluid and electrolyte balance and blood pressure regulation. PMID- 11488774 TI - Pharmacokinetics and pharmacodynamics of NTBC (2-(2-nitro-4-fluoromethylbenzoyl) 1,3-cyclohexanedione) and mesotrione, inhibitors of 4-hydroxyphenyl pyruvate dioxygenase (HPPD) following a single dose to healthy male volunteers. AB - AIMS: NTBC (2-(2-nitro-4-fluoromethylbenzoyl)-1,3-cyclohexanedione) and mesotrione (2-(4-methylsulphonyl-2-nitrobenzoyl)-1,3-cyclohexanedione) are inhibitors of 4-hydroxyphenyl pyruvate dioxygenase (HPPD). NTBC has been successfully used as a treatment for hereditary tyrosinaemia type 1 (HT-1), while mesotrione has been developed as an herbicide. The pharmacokinetics of the two compounds were investigated in healthy male volunteers following single oral administration. The aim of the NTBC study was to assess the bioequivalence of two different formulations and to determine the extent of the induced tyrosinaemia. The mesotrione study was performed to determine the magnitude and duration of the effect on tyrosine catabolism. Additionally, the urinary excretion of unchanged mesotrione was measured to assess the importance of this route of clearance and to help develop a strategy for monitoring occupational exposure. METHODS: A total of 28 volunteers participated in two separate studies with the compounds. In the first study, the relative bioavailability of NTBC from liquid and capsule formulations was compared and the effect on plasma tyrosine concentrations measured. In the second study the pharmacokinetics of mesotrione were determined at three doses. Plasma tyrosine concentrations were monitored and the urinary excretion of mesotrione and tyrosine metabolites was measured. RESULTS: Both compounds were well tolerated at the dose levels studied. Peak plasma concentrations of NTBC were rapidly attained following a single oral dose of 1 mg x kg(-1) body weight of either formulation and the half-life in plasma was approximately 54 h. There were no statistical differences in mean (+/- s.d.) AUC(0,infinity) (capsule 602 +/- 154 vs solution 602 +/- 146 microg x ml(-1) h) or t1/2 (capsule 55 +/- 13 vs solution 54 +/- 8 h) and these parameters supported the bioequivalence of the two formulations. Mesotrione was also rapidly absorbed, with a significant proportion of the dose eliminated unchanged in urine. The plasma half-life was approximately 1 h and was independent of dose and AUC(0,infinity) and Cmax increased linearly with dose. Following administration of 1 mg NTBC x kg(-1) in either formulation, the concentrations of tyrosine in plasma increased to approximately 1100 nmol x ml(-1). Concentrations were still approximately 8 times those of background at 14 days after dosing, but had returned to background levels within 2 months of the second dose. Administration of mesotrione resulted in an increase in tyrosine concentrations which reached a maximum of approximately 300 nmol x ml(-1) following a dose of 4 mg x kg(-1) body weight. Concentrations returned to those of background within 2 days of dosing. Urinary excretion of tyrosine metabolites was increased during the 24 h immediately following a dose of 4 mg mesotrione x kg(-1), but returned to background levels during the following 24 h period. CONCLUSIONS: NTBC and mesotrione are both inhibitors of HPPD, although the magnitude and duration of their effect on tyrosine concentrations are very different. When normalized for dose, the extent of the induced tyrosinaemia after administration of NTBC and over the duration of these studies, was approximately 400 fold greater than that following administration of mesotrione. The persistent and significant effect on HPPD following administration of NTBC make it suitable for the treatment of patients with hereditary tyrosinaemia type 1 (HT-1), whilst the minimal and transient effects of mesotrione minimize the likelihood of a clinical effect in the event of systemic exposure occurring during occupational use. PMID- 11488775 TI - Use of selective serotonin reuptake inhibitors and risk of developing first-time acute myocardial infarction. AB - AIMS: Selective serotonin reuptake inhibitors (SSRIs) have been associated with serotonin depletion in platelets, potentially leading to abnormal aggregation and prolonged bleeding time. In view of the importance of serotonin in coronary thrombosis, and decreased platelet serotonin concentrations associated with SSRIs, the present study was performed to test the hypothesis of a decreased risk of acute myocardial infarction (AMI) associated with SSRIs. METHODS: We conducted a population-based case-control analysis using the UK General Practice Research Database (GPRD). A total of 3319 patients aged 75 years or younger free of clinical conditions predisposing to ischaemic heart disease, with a first-time diagnosis of AMI between 1992 and 1997, and 13 139 controls without AMI matched to cases for age, sex, general practice attended, and calendar time were included. Conditional logistic regression was used to estimate relative risks. RESULTS: Adjusted odds ratios (with 95% CI) for current use of SSRIs, non-SSRIs, or other antidepressants, compared to the group of nonusers of antidepressants were 0.9 (95% CI 0.5,1.8), 0.9 (95% CI 0.7,1.2), and 1.3 (95% CI 0.6,2.8), respectively. As compared with nonuse of SSRIs, current use (regardless of any other antidepressants used) resulted in an adjusted OR of 1.1 (95% CI 0.7,1.6). CONCLUSIONS: The current analysis provides evidence that SSRI exposure does not substantially decrease the risk of developing first-time AMI in patients free of factors predisposing to ischaemic heart disease. However, due to relatively small numbers of exposed subjects and the resulting wide confidence intervals, further studies may be needed to document a lack of effect of SSRIs in subjects without pre-existing diseases predisposing to AMI. PMID- 11488776 TI - Cost of prescribed NSAID-related gastrointestinal adverse events in elderly patients. AB - AIMS: It is well established that nonsteroidal anti-inflammatory drugs (NSAIDs) have gastrointestinal (GI) side-effects. However, the cost of health care resources spent on preventing and managing these side-effects is not clear. The objective of this study was to estimate the direct cost of NSAID-related GI events in an elderly population. METHODS: From the Regie de l'assurance-maladie du Quebec (RAMQ) database, we obtained medical, pharmaceutical and demographic records of a 10% random sample (n = 49 033) of seniors who, between January 1, 1993 and December 31, 1997, had a dispensed prescription of a NSAID. Patients who did not have any GI events during the year prior to their first dispensed prescription were included in the cohort. All patients were followed-up for 2 years. The daily direct Canadian dollar costs of GI events that were incurred by these patients while they were on NSAID therapy were compared with those of GI events that were incurred by these same patients while they were not on NSAID therapy. The difference in these daily costs was attributed to NSAIDs. RESULTS: A total of 12 082 new NSAID users were included in the study. Two hundred and seventeen (1.8%) were hospitalized for GI-related problems; of these, 130 (60%) had their GI hospitalization as their first GI event; 3257 (27.0%) used gastroprotective agents (GPAs), and 857 (26.3%) took GPAs without any apparent prior GI symptoms; 801 (6.6%) had GI diagnostic tests; and 661 (5.5%) died. The average direct costs of GI side-effects per patient-day on NSAIDs were 3.5 times higher than those of a patient-day not on NSAIDs. The direct cost of GI side effects per patient-day on NSAIDs was $1.34, of which more than 70% ($0.94) was attributed to GI events resulting from NSAID treatment. CONCLUSIONS: Approximately one Canadian dollar was added to patient costs for every day he/she was on NSAID therapy. Safer therapies and appropriate patient risk management may potentially reduce NSAID-related health care resource use. PMID- 11488777 TI - A pharmacoeconomic analysis of the impact of therapeutic drug monitoring in adult patients with generalized tonic-clonic epilepsy. AB - AIMS: To carry out a retrospective pharmacoeconomic analysis of the impact of therapeutic drug monitoring (TDM) in adult patients with generalized tonic-clonic epilepsy in an academic, non profit making organization. METHODS: Twenty-five patients who had undergone TDM were compared with 25 age, disease and duration of drug therapy matched controls who had not undergone TDM. Only direct costs were calculated. These included cost to the hospital of providing the TDM service, cost to the hospital per seizure saved, and cost to the patient per seizure saved. RESULTS: Patients undergoing TDM had much more effective seizure control (P = 0.00032, OR 4.846, 95% confidence interval 1.29,18.3), fewer adverse events, better earning and were more likely to be married than the control group. CONCLUSIONS: In patients with adult onset epilepsy, a minimum of two drug estimations per year offers significant benefit in terms of better seizure control, fewer adverse events and greater chances of remission. PMID- 11488778 TI - First results from an intensified monitoring system to estimate drug related hospital admissions. AB - AIMS: An intensified monitoring system was set up to identify drug related hospital admissions and estimate population-based incidences for commonly prescribed medications. METHODS: Pharmacovigilance-centres systematically screened nonelective admissions to emergency rooms or departments of internal medicine for drug related hospitalizations (DRH). Clinical pharmacologists used standardized causality assessment. Service areas of each acute care hospital were defined by 5 digit postal codes that covered 60% of all admissions. Drug dispensing information was available through claims processed by regional pharmacy computing centres. Quarterly incidences were estimated by dividing the number of events by the number of treated patients. RESULTS: 435 DRHs were reported during five quarters. The incidence of ADRs leading to admissions varied for specific drug groups from 1.5/10 000 treated patients to 24/10 000. Quarterly variation of incidences was moderate except for insulin and calcium antagonists. 95% confidence intervals overlap for all quarters within each group. Incidences are sensitive to changes in the definition of the source population. CONCLUSIONS: Our pharmacovigilance monitoring system allows comparisons of population-based incidences of drug-related hospitalizations among drugs and over time. It provides important information for risk management and monitoring outcomes of pharmaceutical quality management programmes. PMID- 11488779 TI - Effects of supratherapeutic doses of ebastine and terfenadine on the QT interval. AB - AIMS: The objective of this study was to compare the effects of high doses of ebastine with terfenadine and placebo on QTc. METHODS: Thirty-two subjects were randomly assigned to four treatments (ebastine 60 mg x day(-1), ebastine 100 mg x day(-1), terfenadine 360 mg x day(-1), placebo) administered for 7 days. Serial ECGs were performed at baseline and day 7 of each period. QT interval was analysed using both Bazett (QTcB) and Fridericia (QTcF) corrections. RESULTS: Ebastine 60 mg (+ 3.7 ms) did not cause a statistically significant change in QTcB compared with placebo (+ 1.4 ms). The mean QTcB for ebastine 100 mg was increased by + 10.3 ms which was significantly greater than placebo but was significantly less (P < 0.05) than with terfenadine 360 mg (+ 18.0 ms). There were no statistically significant differences in QTcF between ebastine 60 mg ( 3.2 ms) or ebastine 100 mg (1.5 ms) and placebo (-2.1 ms); although terfenadine caused a 14.1 ms increase which was significantly different from the other three treatments. The increase in QTcB with ebastine most likely resulted from overcorrection of the small drug-induced increase in heart rate. CONCLUSIONS: Ebastine at doses up to five times the recommended therapeutic dose did not cause clinically relevant changes in QTc interval. PMID- 11488780 TI - Effect of acid secretion blockade by omeprazole on the relative bioavailability of orally administered furazolidone in healthy volunteers. AB - AIMS: The administration of omeprazole may interfere with the absorption of orally administered drugs by reducing gastric pH and hence tablet dissolution. The aim of this study was to investigate the effects of a 5 day administration of omeprazole on the pharmacokinetics of furazolidone. METHODS: Eighteen healthy (nine male and nine female) volunteers were selected. The study had an open randomized two-period crossover design with a 21 day washout period between the phases. Serum concentrations of furazolidone were measured by reversed-phase h.p.l.c. with ultraviolet detection. RESULTS: Administration of omeprazole caused a significant reduction of Cmax [0.34 microg x ml(-1) (range 0.25-0.43) vs 0.24 microg x ml(-1) (range 0.15-0.34)] with no significant delay in absorption tmax [2.5 h (range 1.85-3.0) vs 2.4 h (range 2.06-2.71)]. CONCLUSIONS: Furazolidone was rapidly absorbed after oral administration. Short-term treatment with omeprazole did alter the relative bioavailability of this drug, probably through an effect on absorption kinetics or first-pass metabolism. PMID- 11488781 TI - A double masked placebo controlled study on the effect of nifedipine on optic nerve blood flow and visual field function in patients with open angle glaucoma. AB - AIMS: To investigate whether nifedipine affects ocular perfusion or visual fields in open angle glaucoma patients. METHODS: In a parallel group study nifedipine or placebo was administered for 3 months (n = 30). Ocular fundus pulsation amplitude (FPA), cup blood flow (Flowcup) and visual field mean deviation (MD) were measured. RESULTS: Five patients receiving nifedipine discontinued due to adverse events. Nifedipine did not affect FPA [difference: 0.3 microm (95% CI -0.3,0.9); P = 0.70], Flowcup: [difference: -9 rel.units (95% CI -133,114); P = 0.99], or MD [difference: 0.2dB (95% CI -2.2,2.7); P = 0.51] vs placebo. CONCLUSIONS: Systemic nifedipine is not well tolerated in glaucoma patients and exerts no effect on visual fields or ocular perfusion. PMID- 11488782 TI - Tacrolimus/cerivastatin interaction study in liver transplant recipients. PMID- 11488783 TI - Effect of grapefruit juice on the disposition of omeprazole. PMID- 11488784 TI - Doctors and the law--a personal view. PMID- 11488785 TI - Risk management. PMID- 11488786 TI - Clinical and economic evaluation of surgical treatments for faecal incontinence. AB - BACKGROUND: Faecal incontinence affects 1-2 per cent of the adult population. While many patients can be managed successfully with conservative therapy, a small proportion require surgery. Improved imaging techniques and technological advances have led to the availability of a wide range of surgical treatments. Decision-makers increasingly require clinical and cost-effectiveness studies of surgical treatments for faecal incontinence. This review examines the practical aspects of undertaking such studies. METHODS: The practical issues related to different aetiologies, different types of treatment, defining outcomes, the hidden costs of the condition and its treatment, the rapid changes in technology and issues of patient choice were all considered. A Medline search was undertaken to identify relevant publications, and the reference lists of identified papers were scanned manually. RESULTS: There are few randomized controlled studies and those that have been performed have been limited in their scope. There has also been very limited health economic analysis undertaken. Strategies for conducting such studies, and the criteria they use, have been outlined. CONCLUSION: Randomized trials have a limited role in this setting because of variations in aetiology, difficulty in standardizing procedures, continuing evolution of devices, small patient numbers, concerns for patient choice and the need for long term follow-up. Issues to be addressed when evaluating interventions for faecal incontinence include choosing appropriate measures of surgical outcome, using new continence scoring systems and tools for quality-of-life assessment, and choosing appropriate cost perspectives and time horizons for economic evaluation. PMID- 11488787 TI - Anorectal injury following pelvic radiotherapy. AB - BACKGROUND: Current knowledge of the effects of radiation on the anorectum is based on a limited number of studies. Variability in delivery techniques, both currently and historically, combined with a paucity of prospective and randomized studies makes interpretation of the literature difficult. This review presents the existing evidence and identifies areas that require further work. METHODS: This review is based on a literature search (Medline and PubMed) and manual cross referencing. RESULTS AND CONCLUSION: More than three-quarters of patients receiving pelvic radiotherapy experience acute anorectal symptoms and up to one fifth suffer from late-phase radiation proctitis. About 5 per cent develop other chronic complications, such as fistula, stricture and disabling faecal incontinence. The risk of rectal cancer may be increased. Conservative treatment options are of limited value. Surgery may be considered if symptoms are severe, provided sphincter function is adequate and recurrent disease is excluded. Large prospective studies with accurate dosimetric data and long-term follow-up are needed to provide meaningful information on which to base new strategies to minimize the side-effects from radiotherapy. PMID- 11488788 TI - Randomized clinical trial of stapled versus Milligan-Morgan haemorrhoidectomy. AB - BACKGROUND: The introduction of a stapling technique for the treatment of haemorrhoids has the potential for less postoperative pain, a short operating time and an early return to full activity. The outcome of stapled haemorrhoidectomy was compared with that of current standard surgery in a randomized controlled study. METHODS: Two hundred patients were randomized to either stapled haemorrhoidectomy (n = 100) or Milligan-Morgan haemorrhoidectomy (n = 100) between March 1997 and December 1998. Each patient received standardized postoperative analgesic and laxative regimens, and completed a linear analogue pain score every 6 h during the first day after operation, after the first motion and daily until the end of the first week. Operating time, frequency of postoperative analgesic intake, hospital stay, time to return to normal activity and postoperative complications were also recorded. RESULTS: The mean(s.d.) age of patients in the stapled and surgical groups was 44.1(3.2) and 49.1(12.2) years respectively. The stapled group had a shorter operating time, less frequent postoperative analgesia intake, shorter hospital stay and earlier return to normal activity. Early and late complications, and functional outcome were better in the stapled group. CONCLUSION: Use of a circular stapler in the treatment of haemorrhoidal disease was safe, and was associated with fewer complications than conventional haemorrhoidectomy. PMID- 11488789 TI - Effect of Helicobacter pylori eradication on the ulcer recurrence rate after simple closure of perforated duodenal ulcer: retrospective and prospective randomized controlled studies. AB - BACKGROUND: The possible advantage of eradication of Helicobacter pylori in patients with perforated duodenal ulcer is unknown. This study was planned to assess the prevalence of H. pylori after simple closure of a perforated duodenal ulcer and to study the effect of H. pylori eradication on ulcer persistence and recurrence. METHODS: Some 202 patients were followed prospectively for 2 years after simple closure of a perforated duodenal ulcer (prospective group). A second group of 60 patients was reviewed 5 years or more after perforation closure (retrospective group). The prevalence of H. pylori in patients with perforated duodenal ulcer was compared with that in controls. Patients in the prospective group were randomized to receive either ranitidine alone or quadruple therapy (ranitidine, colloidal bismuth subcitrate, metronidazole and tetracycline) after operation. The incidence of H. pylori infection after the two treatments and the association with residual or recurrent ulcer were studied. In the retrospective group long-term ulcer recurrence was correlated with H. pylori status. RESULTS: The prevalence of H. pylori in patients with perforated duodenal ulcer was not significantly different from that in controls. At every interval of follow-up in the prospective group and in the retrospective group the H. pylori infection rate was significantly higher in patients who had recurrent or residual ulcers. CONCLUSION: Eradication of H. pylori after simple closure of a perforated duodenal ulcer should reduce the incidence of residual and recurrent ulcers. PMID- 11488790 TI - Randomized study comparing cardiac response in endovascular and open abdominal aortic aneurysm repair. AB - BACKGROUND: The aim was to compare the cardiac response and the incidence of adverse cardiac events during and after endovascular (EVAR) and open (OR) repair of abdominal aortic aneurysms (AAAs). METHODS: Seventy-six patients with an AAA suitable for EVAR, and in a general condition allowing open surgery were randomized to EVAR (57 patients) or OR (19 patients). The analysis was on an intention-to-treat basis. Haemodynamic variables were assessed intraoperatively before, during and after aortic occlusion. During the procedure myocardial ischaemia was identified with use of electrocardiography (ECG) and transoesophageal echocardiography (TEE). After operation, cardiac complications were diagnosed by clinical observation, 12-lead ECG at 1 h, 1 day and 7 days, echocardiography at 1 month and measurement of cardiac enzymes. RESULTS: After aortic occlusion, a greater decrease in systemic vascular resistance compared with baseline was observed with OR than with EVAR (- 396 and - 70 dyne s/cm5 respectively; P = 0.03). The stroke work index, as a direct measure of myocardial performance, demonstrated a decrease during OR and an increase during EVAR during aortic occlusion (- 6.6 and + 1.7 g m/m2 respectively; P = 0.03) as well as after aortic occlusion (- 7.6 and + 3.4 g m/m2 respectively; P < 0.01), compared with baseline. The incidence of postoperative clinical cardiac complications was comparable in the two study groups; however, myocardial ischaemia, as observed by ECG and TEE, was observed more frequently in the OR group (ten of 19 versus 15 of 57 patients; P = 0.05). CONCLUSION: Haemodynamic changes were less severe and there was a lower incidence of myocardial ischaemia during EVAR than during OR. Studies are needed to demonstrate whether this may reduce the operative mortality rate. PMID- 11488791 TI - Randomized double-blind controlled trial of roxithromycin for prevention of abdominal aortic aneurysm expansion. AB - BACKGROUND: Macrolide treatment has been reported to lower the risk of recurrent ischaemic heart disease. The influence of macrolides on the expansion rate of abdominal aortic aneurysms (AAAs) remains unknown. The aim was to investigate the effect of roxithromycin on the expansion rate of small AAAs. METHODS: A total of 92 subjects with a small AAA were recruited from two populations. One population consisted of 6339 men aged 65-73 years who were offered a hospital-based mass screening programme for AAA. From this population 66 subjects were recruited. The remaining 26 men were recruited from among 49 subjects diagnosed at interval screening for an initial aortic diameter between 25 and 29 mm. Subjects were randomized to receive either oral roxithromycin 300 mg once daily for 28 days or matching placebo, and followed for a mean of 1.5 years. RESULTS: During the first year the mean annual expansion rate of AAAs was reduced by 44 [corrected] per cent in the intervention group (1.56 mm per year), compared with 2.80 mm per year following placebo (P = 0.02). During the second year the difference was only 5 per cent [corrected]. Multiple linear regression analysis showed that roxithromycin treatment and initial AAA size were significantly related to AAA expansion when adjusted for smoking, diastolic blood pressure and immunoglobulin A level of 20 or more [corrected]. Logistic regression analysis confirmed a significant difference in expansion rates above 2 mm annually between the intervention and placebo groups: odds ratio = 0.09 (95 per cent confidence interval 0.01-0.83) [corrected]. CONCLUSION: In comparison to placebo, roxithromycin 300 mg daily for 4 weeks reduced the expansion rate of AAAs. PMID- 11488792 TI - Non-operative management of haemobilia. AB - BACKGROUND: The aim was to evaluate a non-operative approach to the management of haemobilia. METHODS: This was a retrospective analysis of patients presenting over 10 years with haemobilia. All patients had upper gastrointestinal endoscopy, abdominal ultrasonography and digital subtraction angiography. Superselective coil and/or Gelfoam embolization was done as close as possible to the bleeding site. Completion angiography was performed routinely to confirm adequate embolization. RESULTS: There were 23 patients with liver trauma and six with inflammatory conditions. All patients required resuscitation with fluids and blood transfusion, and had the haemobilia controlled successfully by angiographic embolization. There was one death from fulminant hepatic sepsis. CONCLUSION: This series attests to the efficacy of a non-operative approach to haemobilia using radiological diagnosis and intervention. PMID- 11488793 TI - Experience with laparoscopic ultrasonography for defining tumour resectability in carcinoma of the pancreatic head and periampullary region. AB - BACKGROUND: Computed tomography (CT) is currently the most widely available staging investigation for pancreatic tumours. However, the accuracy of CT for determining tumour resectability is variable and can be poor. Laparoscopic ultrasonography (LUS) is potentially a more accurate method for disease staging. The authors' experience of LUS for staging carcinoma of the pancreatic head and periampullary region is described. METHODS: Fifty-one patients with potentially resectable pancreatic tumours defined at CT underwent further investigation with LUS. Twenty-seven patients subsequently had an open laparotomy. The evaluations of tumour resectability at CT and LUS were compared with the operative findings. RESULTS: At LUS, 24 patients were considered to have resectable tumours, 21 non resectable tumours and six patients were shown to have no pancreatic tumour mass. Twenty-two patients deemed to have a resectable tumour at LUS underwent surgery. Twenty patients were confirmed to have resectable disease and two patients had non-resectable disease. A further five patients underwent surgery. In all five the ultrasonographic diagnosis was confirmed at surgery (four patients with non resectable disease and no pancreatic tumour in one patient). LUS prevented unnecessary extensive surgery in 53 per cent of patients. For the 22 patients who underwent surgery for potentially resectable disease, the positive predictive value of LUS for defining tumour resectability was 91 per cent. CONCLUSION: LUS is an accurate additional investigation for defining tumour resectability and directing management in patients with potentially resectable carcinoma of the pancreatic head or periampullary region. PMID- 11488794 TI - Liver regeneration after major hepatectomy for biliary cancer. AB - BACKGROUND: The aim of this study was to evaluate serial changes in liver volume after major hepatectomy for biliary cancer and to elucidate clinical factors influencing liver regeneration. METHODS: Serial changes in liver volume were determined, using computed tomography, in 81 patients with biliary cancer who underwent right hepatic lobectomy or more extensive liver resection with or without portal vein resection and/or pancreatoduodenectomy. Possible factors influencing liver regeneration were evaluated by univariate and multivariate analyses. RESULTS: The remnant mean(s.d.) liver volume was 41(8) per cent straight after hepatectomy. This increased rapidly to 59(9) per cent within 2 weeks, then increased more slowly, finally reaching a plateau at 74(12) per cent about 1 year after hepatectomy. The regeneration rate within the first 2 weeks was 16(8) cm3/day and was not related to the extent of posthepatectomy liver dysfunction. On multivariate analysis, the extent of liver resection (P < 0.001), body surface area (P = 0.02), combined portal vein resection (P = 0.024) and preoperative portal vein embolization (P = 0.047) were significantly associated with the liver regeneration rate within the first 2 weeks. In addition, body surface area (P < 0.001) and liver function expressed as plasma clearance rate of indocyanine green (P = 0.01) were significant determinants of final liver volume 1 year after hepatectomy. CONCLUSION: The liver regenerates rapidly in the first 2 weeks after major hepatectomy for biliary cancer. This early regeneration is influenced by four clinical factors. Thereafter, liver regeneration progresses slowly and stops when the liver is three-quarters of its original volume, approximately 6 months to 1 year after hepatectomy. PMID- 11488795 TI - Gender differences in long-term survival of patients with colorectal cancer. AB - BACKGROUND: Significant differences exist in the immunological response to surgery. This raises the possibility that gender differences exist concerning the outcome after curative colorectal cancer resection. METHODS: To study this hypothesis, a database of patients with colorectal cancer was analysed prospectively. RESULTS: Some 894 patients were included, 500 (55.9 per cent) were men and 394 (44.1 per cent) were women. Median follow-up was 54.5 months for the entire group and 63.3 months for survivors. The mean(s.e.m.) patient age was 65.3(0.4) years (women 66.1(0.6), men 64.7(0.5) years; P < 0.05). Women lived significantly longer after cancer resection than men (57.8(1.5) versus 52.0(1.3) months; P < 0.05, log rank 0.009). Disease-free survival was significantly longer in women than in men (51.6(1.7) versus 46.0(1.4) months; P < 0.05). Subgroup analysis revealed significant gender differences in Union Internacional Contra la Cancrum (UICC) stages I (n = 195, log rank 0.01) and UICC IV (n = 38, log rank 0.021). Survival analysis after rectal cancer resection revealed significant advantages for women (log rank 0.02), while no gender differences were detected when comparing patients after resection for colonic cancer. Moreover, patients older than 50 years (n = 635) showed significant gender-related survival differences (log rank 0.015). CONCLUSION: Significant gender differences following curative rectal cancer resection were observed. In women disease-free and overall survival were significantly longer. Whether or not these gender differences are related to gender-specific immune functions or to other gender related local or systemic factors remains to be determined. PMID- 11488796 TI - Road traffic fatalities in Victoria, Australia and changes to the trauma care system. AB - BACKGROUND: The aim was to identify organizational and clinical errors in the management of road traffic fatalities and to use this information to improve Victoria's trauma care system. METHODS: A multidisciplinary committee evaluated the complete ambulance, hospital and autopsy records of 559 consecutive road traffic fatalities, who were alive on arrival of ambulance services, in five substantial time periods between 1992 and 1998. Patients who survived more than 30 days were excluded. Errors or inadequacies in each phase of management, including those contributing to death, were identified and an assessment was made of the potential preventability of death. RESULTS: Findings between 1992 and 1998 were similar. In 1998, 1672 problems were identified in 110 deaths with 1024 (61 per cent) contributing to death. Eight hundred and forty-two (50 per cent) of the total problems occurred in the emergency department. There were frequent problems in initial patient reception and medical consultation, resuscitation, investigation and assessment (especially of the abdomen and head), and in transfer to the operating theatre or to a higher-level hospital. Victoria's combined preventable and potentially preventable death rate has been unchanged between 1992 and 1998 (34-38 per cent). CONCLUSION: The problems identified led to a Ministerial Taskforce on Trauma and Emergency Services in Victoria as a consequence of which a new trauma system is now being implemented. PMID- 11488798 TI - Effect of oesophagectomy on monocyte-induced apoptosis of peripheral blood T lymphocytes. AB - BACKGROUND: Surgical stress has been reported to induce immunosuppression. The mechanisms giving rise to T-cell dysfunction following surgery are still unclear. The cellular mechanisms behind T-cell dysfunction following surgery were investigated, based on the induction of T-cell apoptosis and downregulation of T cell signalling molecules. METHODS: Peripheral blood T cells were collected and separated before and after surgery in patients who had oesophagectomy, gastrectomy or cholecystectomy, and studied for their ability to produce cytokines, the induction of T-cell apoptosis with terminal deoxynucleotidyl transferase-mediated dUPT-biotin nick end labelling methods, and the expression of T-cell signalling zeta (TCR zeta) molecules with intracellular staining. RESULTS: The increased degree of T-cell apoptosis, downregulation of TCR zeta molecules and impaired cytokine production of T cells were significant on days 1 and 3 after operation in patients who had oesophagectomy, but not after gastrectomy or cholecystectomy. A higher level of T-cell apoptosis was observed in the co-culture with postoperative monocytes than with preoperative monocytes. CONCLUSION: Peripheral blood T cells obtained after oesophagectomy underwent apoptosis that correlated with the downregulation of TCR zeta molecules. Postoperative monocytes induced by surgical stress were able to mediate the T cell apoptosis. PMID- 11488797 TI - Effect of multimodality therapy on circulating vascular endothelial growth factor levels in patients with oesophageal cancer. AB - BACKGROUND: Angiogenesis is critical for tumour growth and metastasis. The switch to the angiogenic phenotype depends on the net balance between positive and negative angiogenic factors released by the tumour. It was hypothesized that patients with oesophageal cancer would express raised serum levels of vascular endothelial growth factor (VEGF) which would return to normal values with neoadjuvant chemoradiotherapy. METHODS: Forty-four patients with oesophageal cancer who were selected for treatment with neoadjuvant chemoradiotherapy had blood samples taken before treatment, during chemoradiotherapy, before operation, on days 1, 3 and 5 after surgery, and 3 months after resection. Serum levels of VEGF were measured. Values were correlated with response to treatment. Controls were patients who were undergoing surgery for non-malignant conditions. RESULTS: Serum VEGF levels were raised in patients with oesophageal cancer compared with age-matched controls (mean 247 versus 1157 pg/ml; P < 0.01). VEGF levels were unaffected by neoadjuvant treatment but fell significantly on the first day after operation (652 versus 1057 pg/ml before operation; P < 0.05). No decrease occurred in control patients. VEGF levels had returned to preoperative levels by day 5. A similar postoperative rise in VEGF levels was seen in the control subjects (1194 pg/ml on day 5 versus 71 pg/ml before operation; P = 0.001). There was no correlation between VEGF level and response to treatment or tumour stage. VEGF levels had decreased significantly at 3 months following tumour resection (594 versus 1558 pg/ml on day 5; P = 0.03). CONCLUSION: VEGF levels are raised in patients with oesophageal cancer and are unaltered by neoadjuvant treatment, suggesting an additional source other than tumour cells for this proangiogenic agent. PMID- 11488799 TI - Role of fundoplication in patient symptomatology after laparoscopic antireflux surgery. AB - BACKGROUND: Symptoms following antireflux surgery are often seen as unavoidable sequelae of the operation. The aims were to determine the frequency of new adverse sequelae following antireflux surgery and the preoperative incidence of similar symptoms. METHODS: Patients undergoing fundoplication underwent prospective assessment of symptoms using a structured interview before and 6 months after surgery. In addition to the presence or absence of symptoms, Visick scores, visual analogue scales and a composite dysphagia score were used. RESULTS: Some 312 patients were evaluated. Antireflux surgery significantly diminished the symptoms of heartburn, epigastric pain, regurgitation, bloating, odynophagia, nausea, vomiting, diet restriction, nocturnal coughing and wheezing. In contrast, there was a significant increase in inability to belch, diarrhoea and increased passage of flatus. The symptoms of dysphagia, postprandial fullness or early satiety and anorexia were not significantly altered by antireflux surgery. There was, however, a group of patients who experienced new or worsened dysphagia after surgery and were more likely to do so if they had no dysphagia before surgery (31 per cent) than if dysphagia was present before surgery (19 per cent). Some 93 per cent of patients were satisfied with the overall outcome of the operation. CONCLUSION: The majority of patients undergoing laparoscopic fundoplication for gastro-oesophageal reflux derive symptomatic benefit and are satisfied with the outcome. Many of the so-called postfundoplication sequelae are present before surgery in many patients. Overall, antireflux surgery does not lead to increased dysphagia or bloating. PMID- 11488800 TI - Chronic pain and quality of life following open inguinal hernia repair. AB - BACKGROUND: The aim was to determine the frequency and characteristics of chronic pain following open inguinal hernia repair and to identify risk factors for its development. METHODS: This was a questionnaire survey of a historical cohort of patients who underwent inguinal hernia surgery in Aberdeen. The sample comprised all patients (n = 351) who underwent surgery between January 1995 and December 1997, and who were alive and resident in Grampian in October 1999. Outcome measures included self-report of pain persisting for more than 3 months after operation. Pain was characterized by means of the McGill Pain Questionnaire, and quality of life was assessed with the Short Form 36 (SF-36). RESULTS: A total of 226 patients (64 per cent) completed the questionnaire, 67 (30 per cent) of whom reported chronic pain. Reported pain was predominantly neuropathic in character. Patients at increased risk of chronic pain were under 40 years old (P < 0.001), had day-case surgery (P = 0.004), had subsequent surgery on the same side (P < 0.005) and recalled pain before operation (P = 0.005). The SF-36 scores were significantly different in the social functioning, mental health and pain dimensions in patients with chronic pain. CONCLUSION: Chronic pain occurred in 30 per cent of patients after open hernia repair, a higher frequency than has been reported previously. Several risk factors were identified and further prospective research is recommended. PMID- 11488801 TI - Sodium hyaluronate enhances colorectal tumour cell metastatic potential in vitro and in vivo (Br J Surg 2001; 88: 246-50). PMID- 11488802 TI - Extensive surgical cytoreduction and intraoperative hyperthermic intraperitoneal chemotherapy in patients with pseudomyxoma peritonei (Br J Surg 2001; 88: 458 63). PMID- 11488804 TI - Need for secondary interventions after endovascular repair of abdominal aortic aneurysms. Intermediate-term follow-up results of a European collaborative registry (EUROSTAR) (Br J Surg 2000; 87: 1666-73). PMID- 11488806 TI - Endoscopic ultrasonography in the evaluation of idiopathic acute pancreatitis (Br J Surg 2000; 87: 1650-5). PMID- 11488808 TI - Randomized clinical trial of laparoscopic versus open appendicectomy (Br J Surg 2001; 88: 200-5) . PMID- 11488809 TI - Randomized clinical trial of laparoscopic versus open appendicectomy (Br J Surg 2001; 88: 200-5). PMID- 11488811 TI - Role of surgery in mild primary hyperparathyroidism in the elderly, Letter 2 (Br J Surg 2001; 88: 595). PMID- 11488812 TI - The enterohaemorrhagic Escherichia coli (serotype O157:H7) Tir molecule is not functionally interchangeable for its enteropathogenic E. coli (serotype O127:H6) homologue. AB - A major virulence determinant of enteropathogenic Escherichia coli (EPEC) is the Tir molecule that is translocated into the plasma membrane where it orchestrates cytoskeletal rearrangements. Tir undergoes several phosphorylation events within host cells, with modification on a tyrosine essential for its actin-nucleating function. The EHEC (serotype O157:H7) Tir homologue is not tyrosine phosphorylated implying that it uses an alternative mechanism to nucleate actin. This is supported in this study by the demonstration that EHEC Tir is unable to functionally substitute for its EPEC homologue. Like EPEC, the EHEC Tir molecule is phosphorylated within host cells, with the actin-nucleating dysfunction correlated to an altered modification profile. In contrast to EHEC Tir, the EPEC Tir molecule mediated actin nucleation whether delivered into host cells by either strain. Thus, it would appear that EHEC encodes specific factor(s) that facilitate the correct modification of its Tir molecule within host cells. Domain swapping experiments revealed that the N-terminal, alpha-actinin binding, Tir domains were functionally interchangeable, with both the actin-nucleating dysfunction and altered modification profiles linked to the EHEC C-terminal Tir domain. This tyrosine-independent modification process presumably confers an advantage to EHEC O157:H7 and may contribute to the prevalence of this strain in EHEC disease. The presented data are also consistent with EPEC and EHEC sharing non-phosphotyrosine phosphorylation event(s), with an important role for such modifications in Tir function. An EHEC-induced phosphotyrosine dephosphorylation activity is also identified. PMID- 11488813 TI - Phenotypic changes associated with deletion and overexpression of a stage regulated gene family in Leishmania. AB - The LmcDNA16 locus of Leishmania major contains three highly related genes HASPA1, HASPA2 and HASPB, encoding hydrophilic, acylated surface proteins and a tandem pair of unrelated sequences, SHERP1 and SHERP2, coding for a small, hydrophilic protein that localizes to the endoplasmic reticulum and outer mitochondrial membrane. Differential regulation of these genes results in expression of a subset of the HASP proteins and SHERP only in infective stage parasites. To assess the contribution of these molecules to parasite virulence, the diploid LmcDNA16 gene locus has been removed by targeted gene deletion. Homozygous null mutants have precise deletions of both alleles and exhibit no HASP or SHERP expression. They are at least as virulent as wild-type parasites in macrophage invasion and intracellular survival assays, both in vitro and in vivo. Conversely, null mutants engineered to overexpress the entire LmcDNA16 gene locus are unable to survive within the intramacrophage environment despite their differentiation into infective metacyclic parasites. Both null and overexpressing null parasites show increased sensitivity to complement-mediated lysis, suggesting perturbation of their surface architecture. Avirulence in overexpressing parasites correlates with selective depletion of a specific lipid species, decreased expression of the major surface glycoprotein GP63, but no significant downregulation of the glycoconjugate lipophosphoglycan. PMID- 11488814 TI - Binding of the non-typeable Haemophilus influenzae lipooligosaccharide to the PAF receptor initiates host cell signalling. AB - Non-typeable Haemophilus influenzae (NTHi) invades host cells by binding of the platelet-activating factor (PAF) receptor via lipooligosaccharide (LOS) glycoforms containing phosphorylcholine (ChoP). The effect of NTHi infection on host cell signalling and its role in NTHi invasion was examined. The infection of human bronchial epithelial cells with NTHi 2019 increased cytosolic Ca2+ levels, and the invasion of bronchial cells by NTHi 2019 was inhibited by pretreatment with the cell-permeant intracellular Ca2+ chelator BAPTA-AM (P = 0.022) or thapsigargin (P = 0.016). Cytosolic inositol phosphate (IP) levels were also increased after infection with NTHi 2019 (P < 0.001), but not after infection with isogenic mutants expressing altered LOS glycoforms lacking ChoP. PAF receptor antagonist reduced NTHi 2019-stimulated IP production in a dose dependent manner. NTHi 2019 invasion was inhibited by pertussis toxin (PTX) and the phosphatidylinositol-3-kinase inhibitors wortmannin and LY294002. The less invasive strain NTHi 7502 also initiated IP production, but was unaffected by PAF receptor antagonist or PTX. These data demonstrate that the binding of the PAF receptor by NTHi initiates receptor coupling to a PTX-sensitive heterotrimeric G protein complex, resulting in a multifactorial host cell signal cascade and bacterial invasion. Moreover, the data suggest that NTHi strains initiate cell signalling and invade by different mechanisms, and that invasion mediated by PAF receptor activation is more efficient than macropinocytosis. PMID- 11488815 TI - The Akt/PKB pathway is constitutively activated in Theileria-transformed leucocytes, but does not directly control constitutive NF-kappaB activation. AB - The intracellular protozoan parasites Theileria parva and Theileria annulata transform leucocytes by interfering with host cell signal transduction pathways. They differ from tumour cells, however, in that the transformation process can be entirely reversed by elimination of the parasite from the host cell cytoplasm using a specific parasiticidal drug. We investigated the state of activation of Akt/PKB, a downstream target of PI3-K-generated phosphoinositides, in Theileria transformed leucocytes. Akt/PKB is constitutively activated in a PI3-K- and parasite-dependent manner, as judged by the specific phosphorylation of key residues, in vitro kinase assays and its cellular distribution. In previous work, we demonstrated that the parasite induces constitutive activation of the transcription factor NF-kappaB, providing protection against spontaneous apoptosis that accompanies transformation. In a number of other systems, a link has been established between the PI3-K-Akt/PKB pathway and NF-kappaB activation, resulting in protection against apoptosis. In Theileria-transformed leucocytes, activation of the NF-kappaB and the PI3-K-Akt/PKB pathways are not directly linked. The PI3-K-Akt/PKB pathway does not contribute to the persistent induction of IkappaBalpha phosphorylation, NF-kappaB DNA-binding or transcriptional activity. We show that the two pathways are downregulated with different kinetics when the parasite is eliminated from the host cell cytoplasm and that NF-kappaB dependent protection against apoptosis is not dependent on a functional PI3-K Akt/PKB pathway. We also demonstrate that Akt/PKB contributes, at least in part, to the proliferation of Theileria-transformed T cells. PMID- 11488816 TI - Characterization of the intracellular survival of Mycobacterium avium ssp. paratuberculosis: phagosomal pH and fusogenicity in J774 macrophages compared with other mycobacteria. AB - The phagosomes containing viable pathogenic mycobacteria, such as Mycobacterium (M.) tuberculosis and Mycobacterium avium ssp. avium (M. avium), are known to be limited in their ability to both acidify and fuse with late (but not early) endocytic organelles. Here, we analysed the pH and fusogenicity of phagosomes containing M. avium ssp. paratuberculosis (M. ptb), the causative agent of paratuberculosis in ruminants. Using the murine J774 macrophage cell line, we compared viable and heat-killed M. ptb and, in addition, viable or dead M. avium, as well as two non-pathogenic mycobacteria, Mycobacterium smegmatis and Mycobacterium gordonae. Electron microscopic analysis revealed that M. ptb persisted intracellularly in phagosomes for up to 15 days. The phagosomes containing live M. ptb and M. avium were significantly reduced in their ability to acquire some markers for the endocytic pathway, such as internalized calcein, BSA-gold or the membrane protein Lamp 2. However, they were almost completely accessible to 70 kDa fluorescein isothiocyanate (FITC)-dextran and Lamp 1. Overall, the phagosomes containing dead pathogenic mycobacteria behaved similarly to the ones containing live non-pathogenic mycobacteria in all experiments. Using FITC-dextran in a novel fluorescence-activated cell sorting (FACS)-based method, we could also show that the bulk of endocytic compartments, including phagosomes, were only very mildly acidified to approximately pH 6.3 over at least 72 h in J774 cells infected with live M. ptb and M. avium. In contrast, J774 cells treated with heat-killed M. ptb or BSA-coated latex beads showed substantial acidification of the phagosome/endocytic compartments to a pH value of approximately 5.2. After infection with M. smegmatis and M. gordonae, acidification was initially (1-5 h after infection) inhibited, but increased after longer infection to levels similar to those with dead mycobacteria. PMID- 11488817 TI - Remodelling of the actin cytoskeleton is essential for replication of intravacuolar Salmonella. AB - Maturation and maintenance of the intracellular vacuole in which Salmonella replicates is controlled by virulence proteins including the type III secretion system encoded by Salmonella pathogenicity island 2 (SPI-2). Here, we show that, several hours after bacterial uptake into different host cell types, Salmonella induces the formation of an F-actin meshwork around bacterial vacuoles. This structure is assembled de novo from the cellular G-actin pool in close proximity to the Salmonella vacuolar membrane. We demonstrate that the phenomenon does not require the Inv/Spa type III secretion system or cognate effector proteins, which induce actin polymerization during bacterial invasion, but does require a functional SPI-2 type III secretion system, which plays an important role in intracellular replication and systemic infection in mice. Treatment with actin depolymerizing agents significantly inhibited intramacrophage replication of wild type Salmonella typhimurium. Furthermore, after this treatment, wild-type bacteria were released into the host cell cytoplasm, whereas SPI-2 mutant bacteria remained within vacuoles. We conclude that actin assembly plays an important role in the establishment of an intracellular niche that sustains bacterial growth. PMID- 11488818 TI - Azathioprine for atopic dermatitis. AB - For adults with atopic dermatitis (AD) refractory to topical treatment, the choices of second-line therapy are limited. Furthermore, there are concerns about the long-term safety of treatments such as cyclosporin. Limited open studies suggest that azathioprine may be effective, although controlled trial data is lacking. Nevertheless, many UK dermatologists use azathioprine to treat patients with severe AD, despite the potential risk of serious toxicity. Azathioprine myelotoxicity and drug efficacy are now known to be related to the activity of a key enzyme in azathioprine metabolism, thiopurinemethyltransferase (TPMT). Recently, the facility for TPMT measurement has become more widely available, providing the possibility to optimize the therapeutic effect of azathioprine, yet minimise the risk of toxicity. We review the evidence concerning the use of azathioprine for AD, and have identified 128 cases in eight open studies, including our own prospective trial. Improvement in the majority was noted in seven studies, although objective measures of disease activity were used in only one trial. Measurements of TPMT activity were performed in the two most recent studies only. These data underscore the requirement for a prospective randomised controlled trial, and highlight the need to further investigate the role of TPMT measurement in azathioprine usage. PMID- 11488819 TI - Minocycline as a therapeutic option in bullous pemphigoid. AB - We present a retrospective analysis of patients with bullous pemphigoid (BP) treated with minocycline in the Department of Dermatology, Churchill Hospital, Oxford between July 1986 and May 2000. More than 200 patients with BP were seen in clinic during the review period. Of these, 22 patients were treated with minocycline, mostly as adjuvant therapy. The response to treatment was assessed by clinical improvement and whether the dose of concurrent immunosuppressive drugs was subsequently reduced. A major response was seen in six patients, a minor response in 11 and no response was seen in five patients. Minocycline was discontinued in four patients because of the occurrence of side-effects. Overall analysis revealed marked clinical benefits of minocycline in BP. However, this is a non-placebo controlled study and should now be followed by a prospective double blind controlled clinical trial. PMID- 11488820 TI - Health-related quality of life among patients with facial acne -- assessment of a new acne-specific questionnaire. AB - The psychosocial effects of facial acne are well accepted but until recently few validated instruments existed which were suitable for use in clinical trials. The aim of this study was to assess measurement characteristics (reproducibility, correlation with acne severity, and sensitivity to detect change after acne therapy) of a new acne-specific quality of life instrument, the Acne-QoL. We found that the Acne-QoL is reliable, valid and able to distinguish differences across severity groups and improvement over 16 weeks of standard therapy. The use of the Acne-QoL should aid physicians in understanding the impact of facial acne on young adults, and may be useful in assessing therapeutic effects in acne clinical trials. PMID- 11488821 TI - Papular acantholytic dyskeratosis of the vulva. AB - We describe two patients with unusual asymptomatic, papular lesions on the vulva, clinically resembling lichen planus, the histology of which revealed unexpected findings of suprabasilar clefting, acantholysis and dyskeratotic cells giving rise to corps ronds and grains together with hyperkeratosis and parakeratosis, features originally associated with a diagnosis of Darier's disease. Focal acantholytic dyskeratosis has been described in a wide variety of inflammatory and neoplastic processes including those involving mucous membranes and has been attributed various diagnostic labels. We feel that the findings in our patients are consistent with a diagnosis of papular acantholytic dyskeratosis of the vulva, a rare condition, which was first described in 1984. PMID- 11488822 TI - Cutaneous larva migrans contracted in England: a reminder. AB - We report a case of cutaneous larva migrans contracted in England. This case serves as a reminder that the lack of travel abroad should not preclude the diagnosis. We discuss clinical presentation and management of cutaneous larva migrans. PMID- 11488823 TI - Juvenile xanthogranuloma as a sequel to Langerhans cell histiocytosis: a report of three cases. AB - We report three children who had multisystem Langerhans cell histiocytosis (LCH) with cutaneous involvement and subsequently developed juvenile xanthogranuloma (JXG). JXG appeared 3--6 years after the initial manifestation of LCH. JXG lesions, which presented as yellowish papules, revealed typical Touton giant cells and were factor XIIIa positive but S100 and CD1a negative. Non-LCH histiocyte disorders, such as JXG, are known to occur as a reaction to a variety of external stimuli such as infection and trauma. It is therefore conceivable that the inflammatory reaction associated with LCH may have precipitated the development of JXG in our patients. Alternatively, one could speculate that this association might be due to a common histogenetic precursor of the cell types involved. PMID- 11488824 TI - Ecthyma gangrenosum without bacteraemia in a leukaemic patient. AB - Ecthyma gangrenosum is a well recognized cutaneous manifestation of Pseudomonas aeruginosa infections in immunocompromised patients. Most cases of ecthyma gangrenosum have been associated with concomitant septicaemia. However, ecthyma gangrenosum rarely develops due to Ps. aeruginosa in the absence of bacteraemia. We report a rare case of a nonsepticaemic form of ecthyma gangrenosum presenting as a large solitary necrotic ulcer in a patient with acute myelogenous leukaemia. A culture from the lesion revealed the presence of Ps. aeruginosa, but the results of repeated blood cultures were negative. Histological examination revealed numerous tiny eosinophilic bacilli in the dermis and panniculus with Gram's stain. PMID- 11488825 TI - Multiple neutrophilic dermatoses in myelodysplastic syndrome. AB - A 72-year-old woman developed three consecutive processes that showed characteristics of different neutrophilic dermatoses. First, she developed a picture resembling granuloma faciale, followed by a Sweet's syndrome-like eruption, and then by a superficial pyoderma gangrenosum. She was later diagnosed with myelodysplastic syndrome. This case demonstrates that neutrophilic dermatoses form a spectrum of entities that do not necessarily occur in isolation. PMID- 11488826 TI - Zosteriform Kaposi's sarcoma. AB - We report the case of a 92-year-old man with lesions of classic Kaposi's sarcoma occurring in a zosteriform pattern. The mechanisms of dermatomal distribution are speculated and cutaneous lesions presenting in a zosteriform pattern are reviewed. PMID- 11488827 TI - Diffuse plane xanthoma in an otherwise healthy woman. AB - We report a case of diffuse plane xanthoma in a 40-year-old otherwise healthy woman. Her disease began 18 years ago as xanthelasma and progressed to involve large areas of her face, neck and trunk. No associated diseases were detected on repeated laboratory testing. PMID- 11488828 TI - Erythema nodosum and Hodgkin's disease. AB - A 27-year-old woman with a 5-month history of recurrent erythema nodosum was found to have Hodgkin's disease. A temporal relationship between the two disorders suggested a causative role of the lymphoma. A review of the literature yielded 15 cases of this association, suggesting that the diagnosis of Hodgkin's disease should be considered in patients with unexplained erythema nodosum. PMID- 11488829 TI - Isolated lichen planus of the toe nails treated with oral prednisolone. AB - We report a case of lichen planus affecting the toe nails without involvement of the skin or finger nails. To our knowledge this is the first time this has been reported. We also discuss the clinical features, histology and treatment of nail lichen planus. PMID- 11488830 TI - Mycobacterium abscessus: a cutaneous infection in a patient on renal replacement therapy. AB - We report a 72-year-old man on haemodialysis who presented with multiple abscesses on his lower legs. Routine bacterial culture of abscess pus was reported as 'sterile' after 48 h, leading to the suspicion of a mycobacterial infection. Skin biopsy taken for mycobacterial microscopy and culture isolated a heavy growth of Mycobacterium abscessus. PMID- 11488831 TI - Carcinoma cuniculatum of the foot assessed by magnetic resonance scanning. AB - We describe a case of carcinoma cuniculatum of the foot assessed by magnetic resonance scanning. The potential use of this imaging technique in the management of this condition is discussed. PMID- 11488832 TI - Neonatal lupus erythematosus: factors which may lead to clinical disease in the foetus even in the absence of disease in the mother. AB - Neonatal lupus erythematosus (NLE) occurs in neonates of mothers who, in almost all cases, have auto-antibodies to the SSA/Ro associated proteins, but who may have no clinical disease. However, only a small percentage of mothers with SSA/Ro antibodies have affected babies, predisposing factors specific to the foetus or neonate (i.e. HLA pattern) and/or fetal maternal interactions have been proposed to be important. We present a mother with a family history of autoimmune disease, but without clinical disease, whose baby developed cutaneous NLE. Autoantibody determinations as well as the HLA-DR/DQ were performed in the mother and baby. Factors other than the HLA-DR/DQ status of the mother appear to be important in determining whether or not the neonate will develop NLE. Auto-antibodies to endogenous antigens common to the mother, transiently expressed developmental antigens, and the isotype specificity of transferred antibodies may be important in determining disease in the baby. PMID- 11488833 TI - Epitope spreading: protection from pathogens, but propagation of autoimmunity? AB - An epitope is an antigenic determinant, or a site on the surface of an antigenic molecule, to which a single antibody binds. Epitope spreading (ES) refers to the development of an immune response to epitopes distinct from, and noncross reactive with, the disease-causing epitope. Diversification, or the ability of the immune system to attack multiple targets on a pathogen has obvious advantages. Here we review some of the evidence regarding its role in autoimmunity, in humans and in animal disease models. We consider the implications of ES on the development of highly specific therapies for autoimmune disease. We stress that pathogenic ES probably occurs in the context of inherent abnormalities in control mechanisms for the prevention of autoimmunity or other genetic predisposing factors. PMID- 11488834 TI - CD40 expression and function on human dermal microvascular endothelial cells: role in cutaneous inflammation. AB - CD40 is a member of the tumour necrosis factor (TNF) cell surface receptor family. It plays an important role in T-cell dependent B-cell functions and in dendritic cell development. It has also been identified as a cell surface receptor on endothelial cells. In this manuscript, we report that human dermal microvascular endothelial cells (HDMEC) express cell surface CD40 and that the expression of CD40 is increased by the proinflammatory cytokines TNF-alpha and interferon (IFN)-gamma. Additionally, we demonstrate that engagement of HDMEC CD40 with its recombinant CD40 ligand augments the induction of E-selectin, but not of intercellular or vascular cell adhesion molecules on the surface of HDMEC. Furthermore, we show that IFN gamma stimulation of HDMEC results in increased binding of Jurkat leucocytes to HDMEC by a CD40--CD154 dependent pathway. This study thus provides evidence that CD40 expression in HDMEC is regulated by proinflammatory cytokines, and that CD40 functions as an important mediator of cutaneous inflammation. PMID- 11488835 TI - Increased antibody levels to desmogleins 1 and 3 after administration of carbamazepine. AB - Desmoglein (Dsg) 1 and Dsg3 are recognized as the autoantigens in pemphigus foliaceus and pemphigus vulgaris. Pemphigus-like syndromes have been reported to occur in individuals after exposure to a variety of drugs, but pemphigus caused by carbamazepine is not common. We found that anti-Dsg1 and anti-Dsg3 antibody titres were increased in three individuals administered carbamazepine. Antibody titres against Dsgs 1 and 3 were measured by enzyme-linked immunosorbent assay (ELISA). Of 42 serum samples (25 patients administered carbamazepine, eight patients administered valproic acid and nine healthy volunteers) tested by ELISA, three patients administered carbamazepine showed positive reactivity against both Dsg1 and Dsg3. The patient with the highest titre against Dsg1 and Dsg3 (the index values of anti-Dsg1 and anti-Dsg3 were 79.3 and 86.4, respectively) was a 23-year-old woman (Case 1). The other two patients with positive reactivity were a 5-year-old boy (Case 2) and a 65-year-old man (Case 3). In addition, indirect immunofluorescence study showed intercellular antibodies to the cell surface of the whole epidermis with a titre of 1 : 64 in Case 1 and 1 : 2 in Cases 2 and 3. However, no skin or mucosal involvement was found in any of these cases. There was no difference in the serum concentrations of carbamazepine between the three positive cases and the 22 negative cases of carbamazepine administration. From these facts, the lack of skin diseases may be explained by relatively low values of anti-Dsg 1 and 3 antibodies in Cases 2 and 3. However, it cannot be excluded that undefined exogenous and/or endogenous factors are involved in an outbreak of pemphigus. Furthermore, these findings might be helpful for preventing susceptible individuals from exposure to the suspect drugs. PMID- 11488836 TI - Bone mineral density of patients with chronic plaque psoriasis. AB - Reduced bone mineral density (BMD), the major risk factor for osteoporotic fracture, has been linked to palmoplantar pustular psoriasis, but no significant studies have examined BMD in chronic plaque psoriasis (CPP). In this study, in patients with severe CPP had their BMD measured at the nondominant hip and lumbar spine using dual energy X-ray absorbtiometry. Ten male and 10 female Caucasian patients were recruited, with a mean age of 47 years (range 20--71 years). There were no significant differences in BMD between patients and controls. However, patients with psoriatic arthropathy in addition to CPP had a significantly lower mean lumbar spine Z-score (- 1.16) than those without arthropathy (+1.38, P = 0.015). Neither previous nor current treatment with systemic steroids, retinoids or methotrexate significantly affected BMD. We found no evidence that patients with CPP, despite risk factors, have a significantly low BMD, although the subgroup with joint involvement appear be at significantly higher risk of osteoporosis and may therefore require preventative treatment. PMID- 11488837 TI - The clinical photography of Herbert Brown: a perspective on early 20th Century dermatology. AB - Herbert Brown was Consultant Dermatologist to the Victoria Infirmary, Glasgow during the first half of the 20th Century. He was a keen amateur photographer, and his large archive of photographs illustrates comprehensively the work of a clinical dermatologist in that era. PMID- 11488838 TI - Possible phototoxicity with subsequent progression to discoid lupus following pantoprazole administration. PMID- 11488839 TI - Cutaneous botryomycosis in an agricultural worker. PMID- 11488840 TI - Fatal toxic epidermal necrolysis associated with hydroxychloroquine. PMID- 11488841 TI - Basal cell carcinoma arising at the umbilicus. PMID- 11488842 TI - Lentigines in psoriatic plaques: are they unique? PMID- 11488843 TI - A new coal tar preparation (Exorex). PMID- 11488845 TI - Erythropoietin. AB - Maintenance of the red cell volume is a fundamental aspect of ensuring oxygen supply to the tissues. The balance between the very dynamic processes of erythropoiesis and erythrocyte loss is precarious and yet normal individuals experience a remarkably constant haematocrit. This is achieved by a very elegant and sensitive homeostatic mechanism which links tissue oxygen delivery to red cell production. The glycoprotein hormone erythropoietin (EPO) is the principle controller of this process. It is now clear that even minor underproduction of EPO will result in anaemia. The most widespread example of this is the anaemia of end-stage renal failure. The pharmacological use of recombinant human EPO (rHuEPO) in this setting is now well established and has had a dramatic impact on the quality of life of patients with renal disease. With the more widespread use of EPO in other clinical conditions and the advent of novel therapeutic approaches, this is an opportune moment to review the physiology and patho physiology of this fascinating and essential hormone. PMID- 11488846 TI - A standardized flow cytometric method for screening paroxysmal nocturnal haemoglobinuria (PNH) measuring CD55 and CD59 expression on erythrocytes and granulocytes. AB - PNH is a disorder of the pluripotent stem cells resulting in a deficient expression of membrane-bound GPI-anchored proteins in different cell types. Several flow cytometric approaches are designed to detect this antigen deficiency. But they all require drawing and testing of normal samples as control. Therefore, in the present study two flow cytometric assays for the detection of CD55 and CD59 deficiency in erythrocytes (REDQUANT CD55/CD59) and granulocytes (CELLQUANT CD55/CD59) are proposed. Precalibrated beads are used to define the cut off between normal and deficient cell populations. The specificity of the tests has been evaluated in healthy blood donors (n=52) resulting in a clear and reproducible cut off (3%) for the normal percentage of GPI-deficient cells. This cut off has been confirmed in leukaemia and lymphoma patients not suspected for developing PNH. The sensitivity has been tested in patients suffering from known PNH (n=23). Both tests performed in combination allowed a reliable detection of PNH in all patients showing antigen deficiencies in both cell types in most patients (20/23). In contrast, the PNH clones in the investigated patients with MDS (4/19) or AA (4/22) were present in granulocytes or erythrocytes, only. This underlines the necessity of analysing erythrocytes as well as granulocytes. Preliminary data regarding a possible correlation between disease activity and percentage of antigen-deficient cells lead to the assumption that haemolytic crises can only be determined on granulocytes whereas deficient erythrocytes disappeared due to complement-mediated lysis of the PNH clone. In conclusion, the combination of the test kits enables the differential diagnosis of PNH clones in a standardized, simple and rapid approach which may have therapeutic consequences. PMID- 11488847 TI - Use of a new HemoCue system for measuring haemoglobin at low concentrations. AB - In many photometers and spectrometers used in routine practice Beer's law (relating absorbance to concentration) fails at low concentrations. A refinement of the HemoCue haemoglobin system (HemoCue AB, Sweden) provides a facility for accurate measurement of haemoglobin at the lower end of the scale. This study confirmed excellent linearity of the photometer response of the HemoCue Plasma/Low Hb photometer to haemoglobin concentration by direct measurement on whole blood samples, thus providing a reliable method when laboratory facilities are limited and where critical patient management may be dictated by the severity of anaemia. Furthermore, measurements on plasma and on samples of urine containing haemoglobin demonstrated the utility of the device for reliable quantification of plasma haemoglobin and haemoglobinuria, thus providing a useful method in investigating moderate or severe intravascular haemolysis. A study of plasma haemoglobin on stored samples of ethylenediamine tetraacetic acid (EDTA) blood showed that a small but measurable degree of lysis occurs after two days at room temperature and after four days when stored at 4 degrees C. This effect on the red cell count will reflect on the reliability of using such blood as a quality control preparation. PMID- 11488848 TI - Automated reticulocyte counting using the Sysmex RAM-1. AB - Reticulocyte counting using the Sysmex RAM-1 was evaluated. The results of 113 samples analysed on the Sysmex RAM-1 were compared with those from both flow cytometry (FCM) and a manual technique. Reticulocyte counting with the Sysmex RAM 1 showed excellent precision, with an overall coefficient of variance (CV) less than 5%, and almost no carry-over. Reticulocyte counts were stable after blood storage for 48 h at 4 degrees C and room temperature (RT). Linearity was demonstrated very well for Sysmex RAM-1 and FCM (rRAM=0.9998, rFCM=0.9974, P < 0.001) when reticulocyte counts were 5 x 109/l-480 x 109/l. Comparisons of methods showed a good relation for reticulocyte counts between Sysmex RAM-1 and manual counting (r=0.9760), FCM and manual counting (r=0.9623), and Sysmex RAM-1 and FCM (r=0.9527). Analysis of receiver operating characteristic curves showed that the true-positive rate (TPR) was 0.95 for Sysmex RAM-1, and 0.83 for FCM; the area under curve was 0.999 and 0.972 for Sysmex RAM-1, 0.990 and 0.900 for FCM. These findings confirm that Sysmex RAM-1 can make counting reticulocytes easier, more accurate and more reproducible. PMID- 11488849 TI - Impairment of erythrocyte viscoelasticity is correlated with levels of glycosylated haemoglobin in diabetic patients. AB - Erythrocytes from diabetic patients exhibit impaired viscoelastic properties when estimated by various methods. We determined erythrocyte filterability through 5 microm pores, in 51 patients with non-insulin-dependent diabetes mellitus, 18 healthy controls, 15 patients with homozygous beta-thalassemia and 15 with beta thalassemia traits. The filtration measurements were made with a Hemorheometer, which uses the "initial flow rate" principle. To determine the Index of Rigidity (IR) of the red blood cells, we measured the passage time of white blood cell free erythrocyte suspensions, 8% per volume, through the filter. Diabetic patients had significantly increased IR in comparison to healthy controls and to patients with beta-thalassemia trait, but not at the level found in patients with homozygous beta-thalassemia. In diabetic patients, a strong correlation between IR and the percentage of glycosylated haemoglobin was found (r=0.737, P < 0.0001), and a weaker one with serum unconjugated bilirubin (r=0.363, P=0.0097) and serum total lipids (r=0.321, P=0.0286). Patients with severe retinopathy also had significantly increased IR, in comparison to those with or without mild retinopathy. Anaemic diabetic patients, especially those with the anaemia of chronic disease, also had significantly increased IR in comparison to non-anaemic diabetics. No correlation between IR, MCV, MCH, MCHC, RDW, RBC morphology, serum LDH or the presence of erythrocyte inclusions after incubation with nitrous sodium solution was found. Our findings suggest that glycosylation of skeletal proteins probably contributes significantly to the increased membrane rigidity of diabetic erythrocytes. PMID- 11488850 TI - Patients with non-immune chronic idiopathic neutropenia syndrome have increased splenic volume on ultrasonography. AB - Clinically detectable splenomegaly is rarely seen in patients with non-immune chronic idiopathic neutropenia syndrome (NI-CINS). Using ultrasound, we estimated splenic volume in 52 NI-CINS patients and 14 age- and sex-matched normal controls by determining the "corrected splenic index" (CSI) from the product of length, width and thickness of the organ expressed in cm3/m2 body surface area. We found that CSI was significantly higher in the group of patients compared to controls (202.8 +/- 82.0 vs. 133.8 +/- 28.1 cm3/m2, P=0.003), and that individual CSI values was inversely correlated with the number of circulating neutrophils (r= 0.5097, P < 0.0001). About 48.1% of the patients had CSI above 190 cm3/m2 body surface, representing the upper 95% confidence limit of values found in the controls. Patients also had increased serum concentrations of pro-inflammatory cytokines and chemokines mainly produced by activated macrophages (IL-1beta, TNF alpha, RANTES and IL-8), as well as increased serum levels of soluble cell adhesion molecules derived from activated endothelium (sE-Selectin, sICAM and sVCAM). We hypothesize that the increased splenic volume in NI-CINS patients may be due to the accumulation of activated macrophages inside the spleen, possibly as the result of an unrecognized low-grade chronic inflammatory process. The nature of such an inflammation is unknown. A study was designed to search for viral or bacterial genomic material in patients' bone marrow stromal macrophages in which the unknown causal agent might be located. PMID- 11488851 TI - Timing of the appearance of multipotential and committed haemopoietic progenitors in peripheral blood after mobilization in patients with lymphoma. AB - The pattern of emergence of multipotential (CFU-A) and committed (CFU-GM and BFU E) progenitor cells in peripheral blood has been examined in patients with Hodgkin's disease and non-Hodgkin's lymphoma. Mobilization protocols used chemotherapy with or without granulocyte colony-stimulating factor (n=8 and n=5, respectively). In all patients, the numbers of CFU-A, CFU-GM and BFU-E peaked simultaneously, rather than sequentially, suggesting that marrow regeneration after these mobilization protocols occurred from progenitors at all stages of differentiation. We conclude that peripheral blood stem cell harvest strategies based on peak values for total progenitor numbers will also capture maximum numbers of multipotential progenitors. However, the variable relationship between CFU-A and CFU-GM numbers suggests that overall progenitor cell numbers can give only a broad estimate of the absolute numbers of multipotential progenitors in an individual harvest. PMID- 11488852 TI - The effects of prior induction therapy with melphalan on subsequent peripheral blood progenitor cell transplantation for myeloma. AB - High dose chemoradiotherapy with autologous peripheral blood progenitor cell transplantation (PBPCT) may improve outcome in myeloma. Melphalan is an effective drug in the treatment of myeloma, but is potentially toxic to progenitor cells. We studied 8 patients receiving intermittent intravenous melphalan (25 mg/m2) as induction therapy before PBPCT to assess engraftment characteristics post transplantation. Comparison was made with an age-matched control group of patients with non-Hodgkins lymphoma who had not received melphalan during induction therapy. There was correlation (P=0.037) between the dose of melphalan per kg body weight given, premobilization, and days to neutrophil engraftment, but no significant difference between the two groups in neutrophil recovery. The study group had delayed platelet recovery (P=0.01) and required more platelet support post-transplantation (P=0.05). 3-4 weekly melphalan (25 mg/m2) up to 6 courses was delivered to patients who went on to PBPCT without significantly influencing neutrophil recovery but with a negative impact on platelet recovery. PMID- 11488853 TI - The PFA-100 system for the assessment of platelet function in normotensive and hypertensive pregnancies. AB - Platelet function was studied in 30 pregnant women: 14 normotensive (C), and 16 affected by pregnancy-induced hypertension (PIH). Platelet aggregometry (PA) on platelet-rich plasma according to Born was compared with the new PFA-100 System (Dade International Inc, Miami, USA). This device evaluates platelet function (expressed in seconds as closure time, CT) in anticoagulated whole blood ex vivo at high shear rates. PA (expressed as percentage of light transmission) and CT were measured at baseline and after incubation with L-Arginine (L-Arg). MANOVA for repeated measures showed that L-Arg incubation significantly decreased PA (F=7.2, P < 0.05) and increased CT (F=6.05, P < 0.05) in the whole population of pregnant women. Moreover, we analysed separately both parameters in C and in PIH subjects. No differences in PA were found in both groups, neither at baseline nor after L-Arginine incubation. In contrast, CT was significantly longer in PIH in comparison to C before (95.9 s vs. 84 s, P < 0.05) as well after (115 s vs. 92 s, P < 0.05) L-Arginine incubation. Data from PFA-100 confirm our previous reports that during pregnancy the L-Arginine: Nitric Oxide pathway regulates platelet function. In hypertensive patients a significant decrease in platelet function was found by using the PFA-100 system. PMID- 11488854 TI - Successful salvage of RAEB/AML relapsing early post allograft with FLAG-Ida conditioned mini-allograft: a report of two cases. AB - Management options are often limited for patients with AML or high grade myelodysplasia (MDS) relapsing within a year of allogeneic transplantation. We report, in two such patients, the use of re-induction with FLAG-Ida chemotherapy, followed by the infusion of GCSF-mobilized blood stem cells from the same HLA matched donor. Both patients achieved durable complete remissions with good quality of life and longer disease-free survival than after the first myeloablative allografts. This mini-allograft approach offers a practical, well tolerated salvage and a potentially curative treatment for relapsed AML/high grade MDS patients failing a first conventional myeloablative allogeneic transplants. PMID- 11488855 TI - Concomitant angioimmunoblastic T-cell lymphoma and low grade B-cell lymphoproliferative disorder. AB - The presence of a rearranged immunoglobulin gene, in addition to the expected T cell receptor gene rearrangement, is a frequent, albeit poorly understood, finding in the setting of angioimmunoblastic lymphadenopathy. A case of an angioimmunoblastic T-cell lymphoma is presented, where this apparently paradoxical dual gene rearrangement could be ascribed to the coexistence of an occult B-cell lymphoproliferative disorder. PMID- 11488856 TI - Treatment of peri-implantitis by local delivery of tetracycline. Clinical, microbiological and radiological results. AB - The purpose of this study was to investigate the clinical, microbiological and radiological effects of peri-implantitis therapy by local delivery of tetracycline. In 25 partially edentulous patients, 30 implants with radiographic evidence of circumferential bone loss, and peri-implant probing depths > or =5 mm were treated with polymeric tetracycline HCl-containing fibers. Clinical and microbial parameters were recorded at baseline, and 1, 3, 6, and 12 months (M) after treatment. Standardized radiographs were obtained at baseline, M3, and one year after treatment. Two patients were discontinued from the study after 180 days because of persisting active peri-implantitis with pus formation. The remaining subjects showed a significant decrease of mean peri-implant probing depth from 6.0 to 4.1 mm (M1, P<0.001), which was maintained over 12 months. In comparison to baseline, the bleeding tendency was significantly reduced after one month, and thereafter (P<0.001). No significant recession of the mucosal margin was noted. The radiologically determined distance from the shoulder of the implant to the bottom of the bony defect decreased slightly, but not significantly, from 5.2 to 4.9 mm. At M1, M3 and M6, mean total anaerobic cultivable bacterial counts were significantly lower than at baseline (P<0.001). A significant decrease in frequency of detection was noted for Prevotella intermedia/nigrescens, Fusobacterium sp., Bacteroides forsythus, and Campylobacter rectus (P<0.01). Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Eikenella corrodens had very low baseline frequencies that could not be significantly suppressed further. In conclusion, therapy of peri-implantitis by local delivery of tetracycline had a positive effect on clinical and microbiological parameters. PMID- 11488857 TI - Clinical and microbiological evaluation of ligature-induced peri-implantitis and periodontitis in dogs. AB - The purpose of this study was to evaluate the attachment loss around teeth and implants by clinical and microbiological analysis. The mandibular premolars were extracted in 5 mongrel dogs and, 3 months later, two titanium implants were installed on each side of the mandible and, after another 3 months, abutment connection was performed. Plaque control in the implants and maxillary premolars was maintained for two weeks prior to the start of the main experiment. On day 0 and 30 days after ligature placement, microbiological samples were obtained and relative attachment level was measured for the teeth and implants. The presence of Porphyromonas gingivalis, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Prevotella nigrescens was evaluated by polymerase chain reaction technique on day 0 and 30 days after ligature placement. None of the above bacteria were detected on day 0. Thirty days after ligature placement, P. gingivalis was present in 95% and 85% and B. forsythus was present in 80% and 85% of the implants and teeth sites, respectively. Statistical analysis (one-way RM-ANOVA) showed a significant difference (P<0.01) between pre- and post-induction measurements around teeth and implants. However, there was no significant difference (P=0.41) in the rate of attachment loss, between periodontitis and peri-implantitis. It can be concluded that: (1) P. gingivalis and B. forsythus were strongly associated with induced peri-implantitis and periodontitis, and (2) induced peri-implantitis and periodontitis presented a similar rate of attachment loss. PMID- 11488859 TI - Clinical evaluation and prosthetic complications of single tooth replacements by non-submerged implants. AB - The aim of the present study was to evaluate single tooth replacements by non submerged implants. In the time period from 1990 to 1998, 72 patients were consecutively admitted for treatment with a total of 109 solid screw ITI-implants supporting single crowns. All crowns were ceramic to metal fused with a ceramic occlusal surface and mounted to the octa-abutment. The mean observation time was 4.3 years, ranging from >1 to 9 years. The implants were monitored regularly by periimplant parameters. Periapical radiographs using the parallel technique were taken after the healing period and in 1999 for comparative measurements. No implant was lost during the healing phase, while 3 failures occurred after a loaded period of 2.5 and >5 years, respectively. The 5-year cumulative survival rate was 99.1%. In the course of the study, only 2 implants exhibited marginal inflammation that required treatment. The radiographically measured distance (DIB) from the implant shoulder to the first implant-bone contact was significantly increased in 1999 and different between the two readings. However, this difference in DIB was not significant between implants with a short (1 year) and long (>5 years) observation period. It was concluded that changes in the crestal bone level occur mostly in the first postsurgical year. Prosthetic complications were rare, mostly encountered in the first year after loading and often limited to re-tightening of the occlusal screw. PMID- 11488858 TI - Removal of HA and TPS implant coatings and fibroblast attachment on exposed surfaces. AB - The removal of implant coatings may be necessary if rough implant surfaces are exposed subgingivally due to progressive peri-implant bone loss or if they are also supragingivally exposed because of progressive gingival recession, thus facilitating plaque formation and impairing tooth cleaning done at home. The aim of this experimental study was to develop diamond-coated files and rubberized polishers for machine-driven instrumentation of implant cylinders, and present an instrumentation concept for the complete removal of rough titanium plasma spray (TPS) and hydroxylapatite (HA) layers and polishing exposed titanium surfaces. The surface structure and its possible contamination by the instrumentation process was investigated using laser profilometry, scanning electron microscopy, and x-ray fluorescence analysis. The effect of impurities on the growth of human fibroblasts was tested in cell culture. The results show that TPS and HA implant coatings can be removed with the modified implant files. After polishing the exposed titanium surfaces, roughness depths of Rz=4.6 microm (TPS) and Rz=5.7 microm (HA) and, for implant cylinders, 3.4 microm were found. During the instrumentation of implant surfaces, contamination with the material of the instrument must be expected. It was shown that, in principle, growth of human gingival fibroblasts on the instrumented surfaces is possible. The cells were intimately associated with one another and, compared to culture controls, demonstrated good adhesion with strict orientation to the microstructure of the scoring left by instrumentation. The biological consequences and mechanisms of cell adhesion on instrumented surfaces require further investigation. PMID- 11488860 TI - Retrospective analysis of the influence of abutment structure design on the success of implant unit. A 3-year controlled follow-up study. AB - A comparative study of 96 Spline and Threadloc implants is presented. The authors examined the possibility of the failure of the retaining screws of two different antirotational systems (the Threadloc system with hexagonal polyhedral extension and the Spline system), and the possible incorrect link between the abutment and the transfer. After an observation period of 36 months starting from the day of prosthesis insertion, three single Threadloc fixtures (20%) and five pairs of joint Threadloc fixtures (5.7%) presented problems and a partial prosthetic screw loosening. For the Spline series fixtures, no screw loosening was encountered. Regarding the safety of the link between abutment and implant, the best results were obtained from the Spline system, which did not hinder the dentist in positioning the link with the abutment. This is the result of the different geometries of the two antirotational mechanisms which are present on the two fixtures. It is believed that the precise contact between the abutment and the fixture avoids the triggering of a phlogistic process, which can develop into periimplantitis. Therefore, when hex systems are used, an x-ray, which verifies this correct situation, is always recommended during the follow-up. PMID- 11488861 TI - Role of chitin beads in the formation of jaw bone by guided tissue regeneration. An experiment in the rat. AB - It has been reported that local application of bone grafts or synthetic bone substitutes (filler materials) may favour bone formation when used in combination with guided tissue regeneration (GTR). Therefore, the aim of the present investigation was to evaluate the effect of application of chitin beads (a bioabsorbable natural polymer) as a bone substitute in bone formation by GTR. The experiment was carried out in 25 rats. The mandibular ramus was exposed on one side after elevation of a muscle-periosteal flap, and a teflon capsule filled with chitin beads (2.0 mm in diameter) was placed with its opening facing the lateral aspect of the ramus. On the contralateral side of the jaw, serving as control, an empty teflon capsule was placed in the same manner. Groups of 5 animals were sacrificed at 7, 15, 30, 60 and 120 days following capsule placement. Histological analysis demonstrated that the amount of newly formed bone was similar in both experimental and control specimens, amounting to approximately 3% of the central/largest, cross-sectional area created by the capsule at 15 days, and to approximately 9% of this area at 30 days following capsule placement. At 60 and 120 days, however, the amount of newly formed bone observed in the control specimens was twice as large as that observed in the test specimens, amounting to approximately 31% of the cross-sectional area created by the capsule at 60 days, and to approximately 45% at 120 days. It is concluded that, although chitin beads (2.0 mm in diameter) are biocompatible, their presence retards bone formation in the model system used. PMID- 11488862 TI - Migration of osteoblastic cells on various guided bone regeneration membranes. AB - To evaluate the biological effects of guided bone regeneration (GBR) barrier materials on osteoblastic cell migration, migration of mouse osteoprogenitor cells (MC3T3-E1) was examined, in vitro, on various membranes. Eight commercially available GBR membranes - bovine type I collagen (BioMend; BM), porcine type I collagen (BioGide; BG), bovine type I atelocollagen (Tissue Guide; TG), polylactic acid (Epi-Guide; EG), co-polymer of polylactic acid and polyglycolic acid (Resolute; RL, Resolut XT; RL-XT), expanded polytetrafluoroethylene (e-PTFE; Gore Tex; GT) and co-polymer of cellulose acetate and nitrocellulose (Millipore filter; MP) - were tested. A 3x5 mm section of the membrane was fixed to the bottom of a culture dish with double-sided adhesive tape, and half of the membrane was closely covered by PARAFILM (American National Can) to leave an unexposed area for cell migration. The border between exposed and unexposed areas was marked as a baseline of cell migration. Membranes were then plated with 3 ml of cell suspension at an initial density of 1x105 cells/ml in alpha-MEM culture medium with 10% fetal bovine serum and ascorbic acid. After a 5-hour incubation, non-attached cells were completely washed out with phosphate buffered saline and the PARAFILM cover was removed. After 3 days cultivation, specimens were fixed with 10% buffered formalin and stained briefly with hematoxylin. The area of cell migration on a membrane was analyzed using a LA 500 Image Analysis System and migration area per unit length of the baseline (mm2/mm) was compared among membranes. Results demonstrated that cell migration was greater in the order: RL>RL-XT, BM, TG, MP>EG, BG. Membranes except for BG, EG and GT showed the migration rate equal to or higher than a plastic culture cover slip (Celldesk) (P<0.01) on which cells generally grow favorably. Only a small number of the cells attached to GT, and the net cell migration for the membrane could not be determined. These results indicate that GBR barrier materials per se may influence the process of bone regeneration in vivo through the effects of their presence on cell migration. PMID- 11488863 TI - Effect of recombinant human bone morphogenetic protein-2 on bone regeneration and osseointegration of dental implants. AB - Recombinant human bone morphogenetic protein-2 (rhBMP-2) induced bone regeneration and osseointegration was evaluated in bony defects created within the hollow chamber of endosseous dental implants in 14 foxhound dogs. Bilateral extractions of mandibular premolars were performed and surgical implantation of 104 hollow cylinder implants followed after 8 weeks of healing. Experimental implants had their hollow chamber filled with 20 microg of rhBMP-2 delivered with a bovine collagen carrier, whereas the control implants had their apical chamber left empty. Dogs were followed for 2, 4, 8 and 12 weeks. Histomorphometric evaluation and immunohistochemical analysis were performed. Minimal bone was regenerated at 2 weeks for both groups. At 4 weeks, bone fill averaged 23.48% for the rhBMP-2 and 5.98% for the control group (P<0.05). At 8 weeks, mean bone fill was 20.94% and 7.75% for the rhBMP-2 and the controls, respectively (P<0.05). At 12 weeks, mean bone fill was 31.39% and 24.31% for the rhBMP-2 and control implants, respectively (P>0.05). Bone-implant contact (BIC) increased for both groups over time and at 8 weeks the rhBMP-2 BIC value was 18.65% and for the control 7.22% (P<0.05). At 12 weeks, the BIC was 43.78% and 21.05% for the rhBMP 2 and the control group, respectively (P<0.05). Immunohistochemical staining for type II collagen was positive only for parts of the collagen carrier and formation of cartilaginous intermediate was not observed in any of the specimens. The results suggest that, in confined defects adjacent to dental implants, rhBMP 2 can induce bone regeneration in close apposition to the implant surface. PMID- 11488864 TI - Early endosseous integration enhanced by dual acid etching of titanium: a torque removal study in the rabbit. AB - Textured implant surfaces are thought to enhance endosseous integration. Torque removal forces have been used as a biomechanical measure of anchorage, or endosseous integration, in which the greater forces required to remove implants may be interpreted as an increase in the strength of bony integration. The purpose of this study was to compare the torque resistance to removal of screw shaped titanium implants having a dual acid-etched surface (Osseotite) with implants having either a machined surface, or a titanium plasma spray surface that exhibited a significantly more complex surface topography. Three custom screw-shaped implant types - machined, dual acid-etched (DAE), and titanium plasma sprayed (TPS) - were used in this study. Each implant surface was characterized by scanning electron microscopy and optical profilometry. One DAE implant was placed into each distal femur of eighteen adult New Zealand White rabbits along with one of the other implant types. Thus, each rabbit received two DAE implants and one each of the machined, or TPS, implants. All implants measured 3.25 mm in diameter x 4.00 mm in length without holes, grooves or slots to resist rotation. Eighteen rabbits were used for reverse torque measurements. Groups of six rabbits were sacrificed following one, two and three month healing periods. Implants were removed by reverse torque rotation with a digital torque measuring device. Three implants with the machined surface preparation failed to achieve endosseous integration. All other implants were anchored by bone. Mean torque values for machined, DAE and TPS implants at one, two and three months were 6.00+/-0.64 N-cm, 9.07+/-0.67 N-cm and 6.73+/-0.95 N-cm; 21.86+/-1.37 N-cm, 27.63+/-3.41 N-cm and 27.40+/-3.89 N-cm; and 27.48+/-1.61 N-cm, 44.28+/-4.53 N-cm and 59.23+/-3.88 N-cm, respectively. Clearly, at the earliest time point the stability of DAE implants was comparable to that of TPS implants, while that of the machined implants was an order of magnitude lower. The TPS implants increased resistance to reverse torque removal over the three-month period. The results of this study confirm our previous results that demonstrated enhanced bony anchorage to dual acid-etched implants as compared to machined implants. Furthermore, the present results indicate that dual acid etching of titanium enhances early endosseous integration to a level which is comparable to that achieved by the topographically more complex TPS surfaces. PMID- 11488865 TI - Bicortical titanium screws for critical orthodontic anchorage in the mandible: a preliminary report on clinical applications. AB - Critical anchorage during orthodontic treatment in the mandible needs both time and effort and patient compliance. In 8 patients, 12 bicortical titanium screws (BIS) were used as anchorage units for orthodontic molar protraction. The criteria for patient selection were: critical anchorage in the lower jaw (i.e. retraction of anterior teeth undesirable) and molar extraction sites. After insertion of the screws in local anesthesia, orthodontic forces were applied immediately. One screw worked loose and had to be removed before the end of treatment. Problems encountered included impingement of the screw head and slight inflammatory reactions of the surrounding mobile mucosa, which necessitated premature removal of two screws. After healing, a new insertion site was chosen. Further treatment was uneventful. Anchorage for orthodontic forces as described offers several advantages. The total treatment time is reduced as the screws can be loaded immediately. The line of action of the orthodontic force coincides with the level of the center of resistance of the molar resulting in a favorable translatory tooth movement. Treatment does not depend on patient cooperation. PMID- 11488866 TI - Mathematical analysis of projection errors in "paralleling technique" with respect to implant geometry. AB - "Standardized" radiographs acquired in paralleling technique serve for monitoring of marginal bone levels around endosseous implants. Under clinical conditions, parallel adjustment of the film to the implant is beset with great difficulties. A mathematical model matching clinical conditions was developed to evaluate projection geometry within an interval of clinically relevant angulations (+/- 10 degrees from parallel position). Radiographs of two implants (Frialit 2, Friadent AG, Mannheim, Germany; Implant No. 1: 3.8 mm, length 10 mm; Implant No. 2: 6.5 mm, length 13 mm) were separately produced per angulation (2 degrees increments) at one focus-object distance (FO=322.9 mm). Implant images were repeatedly measured along their midline/vertical edge, local magnification (MF) was calculated and the values were compared to the computed ones. Projected dimensions of the implants were calculated for a second distance (232.3 mm). The experimentally acquired data were in agreement with the mathematical calculation. MF calculated for assessment along the vertical edge varied less (+/-1.94% from mean value) than along the midline (+/-2.74%), with a range of 1.037-1.068 (FO=322.9 mm) and 1.061-1.099 (FO=232.3 mm) for implant No.1, and 1.060-1.101 (FO=232.3 mm) and 1.037-1.069 (FO=322.9 mm) for Implant No. 2. Magnification revealed a mean variation of 4%. Radiographic evaluation of periimplant bone level should not exceed a precision of 0.5 mm, when parallelism between film and implant is not guaranteed and FO is less than 380 mm. PMID- 11488867 TI - Radioimaging of implants in rats using Tc-99m-MDP. AB - Radiopharmaceutical isotopes are widely used clinically to detect tumors of osteogenic origin. One example is Technetium-99m methylene diphosphonate (Tc-99m MDP). When viewed with a gamma camera, the concentration of the isotope (increased gamma activity) indicates an area of increased bone activity. This technology provides an opportunity to measure bone growth around implants in vivo. The purpose of this study was to measure Tc-99m-MDP activity around titanium alloy implants placed in the tibiae of rats. Some implant sites were treated with a growth factor; other sites served as controls. The hypothesis tested was that implants placed with a growth factor would have greater associated Tc-99m-MDP activity. Twelve adult male Sprague-Dawley rats were anesthetized and surgical access to the medial proximal tibiae was obtained. Titanium alloy screw implants were placed in six animals along with 65 microgram of acidic fibroblast growth factor (FGF-1); the other six animals received implants only and served as controls. After five days, rats were injected with 1500 microCi of Tc-99m-MDP. After 3 hours, rats were imaged with a gamma camera. The Tc-99m-MDP intensity associated with each implant was quantified and the means for each group were compared using ANOVA. Implants treated with FGF-1 demonstrated significantly more Tc-99m-MDP activity than implants alone. This suggests that Tc-99m-MDP analysis may be a useful tool for determining bone growth around implants in laboratory animals in vivo. PMID- 11488868 TI - Histology of human alveolar bone regeneration with a porous tricalcium phosphate. A report of two cases. AB - Porous beta-phase tricalcium phosphate particles (pTCP) (Cerasorb) were used in two patients to restore or augment alveolar bone prior to the placement of dental implants. In one patient, pTCP was used to fill a large alveolar defect in the posterior mandible after the removal of a residual cyst, and in another patient to augment the sinus floor. Biopsies were taken at the time of implant placement, 9.5 and 8 months after grafting, respectively, and processed for hard tissue histology. Goldner-stained histological sections showed considerable replacement of the bone substitute by bone and bone marrow. In the 9.5 months biopsy of the mandible, 34% of the biopsy consisted of mineralised bone tissue and 29% of remaining pTCP, while the biopsy at 8 months after sinus floor augmentation consisted of 20% mineralised bone and 44% remaining pTCP. Bone and osteoid were lying in close contact with the remaining pTCP and were also seen within the micropores of the grafted particles. Tartrate resistant-acid phosphatase (TRAP) multinuclear cells, presumably osteoclasts, were found surrounding, within and in close contact with the pTCP particles, suggesting active resorption of the bone substitute. Remodelling of immature woven bone into mature lamellar bone was also found. No histological signs of inflammation were detected. The limited data presented from these two cases suggest that this graft material, possibly by virtue of its porosity and chemical nature, may be a suitable bone substitute that can biodegrade and be replaced by new mineralising bone tissue. PMID- 11488869 TI - Assessment of accuracy in breast cytology. PMID- 11488870 TI - An audit of "equivocal" (C3) and "suspicious" (C4) categories in fine needle aspiration cytology of the breast. AB - We have audited the frequency of use and outcome of the "equivocal/atypia probably benign" (C3) and "suspicious of malignancy" (C4) category for breast cytology in our Unit. A total of 14 935 cytological specimens were reported by at least one of the three pathologists with a special interest in breast pathology, according to five categories of the NHSBSP guidelines for cytology reporting, 1992; 3.7% (555 cases) and 3.9% (587 cases) of cases were classified as equivocal (C3) and suspicious (C4), respectively, giving a total rate (C3 + C4) of 7.6%. Of the C3 cases, 68% were subsequently benign and 32% were malignant. Of the C4 cases, 19% were subsequently benign and 81% malignant. The commonest benign lesions in both categories were fibroadenomas (7.6% of C3 and 19.8% of C4), fibrocystic change (14.3% of C3 and 12.5% of C4), radial scars (6.2% of C3 and 10.4% of C4) and papillomas (6.2% of C3 and 6.3% of C4). Of the malignant lesions (particularly those classified as C3), a high proportion were low grade or special type cancers. The categories of atypia probably benign (C3) and suspicious of malignancy (C4) in breast cytology provide a strategy for classification of problematic or uncertain cases; this maintains the predictive value of the benign (C2) and malignant (C5) categories, and allows separation of these difficult cases into clinically useful groups with differing probabilities of malignancy. PMID- 11488871 TI - Is there a better way to assess performance in breast cytology? AB - Is there a better way to assess performance in breast cytology? Results generated from a series of breast fine needle aspirates (BrFNA) are semi-qualitative and therefore sensitivity, specificity and accuracy are not necessarily the best measures of effectiveness. This paper considers the likelihood ratio of cancer (LR+), which can provide the probabilities for cancer being present or absent for cancer across the range of diagnostic categories used in the National Health Service Breast Screening Programme (NHSBSP), i.e. C1 to C5. Using LR+, receiver operating characteristic (ROC) curves can be generated and used to compare performance between pathologists, laboratories or years. These are illustrated using test results from 1997 to 1999 for the Derbyshire Royal Infirmary. LR + and ROC are meaningful measures. They could replace the 12 calculations currently used for the evaluation of effectiveness of BrFNA in the National Health Service Breast Screening Programme (NHSBSP) and for validation of pathologists' performance. PMID- 11488872 TI - Rapid screening: a comparative study. AB - Although rapid screening of negative and inadequate cervical smears is a quality assurance requirement for all UK laboratories, there has been little attempt to standardize the method and laboratories make use of a number of different techniques and times. The aim of this study was to assess the sensitivity of these various techniques by measuring their ability to pick out known false negative smears. Completed questionnaires from 123 laboratories across England revealed that 52% of laboratories use a "step" technique, 19% use "turret", 15% use random paths and 34% attempt to rescreen the whole slide quickly. Twenty-two percent of laboratories use a mixture of techniques. Timings are also variable, with the majority of laboratories allowing screeners to review slides at a pace decided by themselves but usually between 1 and 2 min. The study involved 120 participants who performed a total of 24 000 rapid screens. The results showed that, of the 90 abnormal slides used in the study, 62 cases (69%) were identified as abnormal or needing review by more than 50% of participants. Overall rapid screening picked out 58% of high-grade squamous abnormalities, 59% of low-grade abnormalities and 72% of glandular lesions. Step screening performed best, followed by whole slide/random and then turret. One minute was the optimum time and there was a significant fall in performance once individuals attempted to rescreen large numbers (>50). The most significant finding was the marked variation in the performance of individuals using the same slide sets. PMID- 11488873 TI - Invited commentary--Rapid review: current practice. PMID- 11488874 TI - For debate--Ethical considerations of gynaecological liquid-based cytology and human papillomavirus studies. PMID- 11488875 TI - Fine needle aspiration cytology of primary lymphoma of the thyroid: a report of 17 cases. AB - Between 1980 and 1998, 4272 thyroid surgical specimens with a preoperative fine needle aspirate were sent to our Anatomical Pathology Department. Among these cases there were 17 primary thyroid lymphomas, which constituted 0.3% of all the thyroid lesions and 2.3% of the thyroid malignancies. Seven cases were diffuse large B-cell (DLBC) lymphomas and 10 were MALT lymphomas. Of the DLBC lymphomas six were correctly diagnosed by fine needle aspiration cytology (FNAC) and one was diagnosed as positive for malignancy, and among MALT lymphomas four were diagnosed as lymphoma, four as suspicious for lymphoma, and three as Hashimoto's thyroiditis (HT). Our data indicate that the diagnosis of primary thyroid lymphoma of high grade is easy, and immunocytochemistry (ICC) can confirm suspicious cases. The diagnosis of MALT lymphoma is more difficult; ICC can confirm suspicious cases, and false-negative results seem to be caused by sampling error, because HT usually coexists with MALT lymphoma. PMID- 11488876 TI - How would you classify this fine needle aspirate of breast: C2-5? PMID- 11488877 TI - Educational case report. Fine needle aspiration cytology of adenoid cystic carcinoma of the breast. PMID- 11488878 TI - Ibotenate injections into the pre- and parasubiculum provide partial protection against kainate-induced epileptic damage in layer III of rat entorhinal cortex. AB - PURPOSE: A loss of neurons in layer III of the entorhinal cortex (EC) is often observed in patients with temporal lobe epilepsy and in animal models of the disorder. We hypothesized that the susceptibility of layer III of the EC to prolonged seizure activity might be mediated by excitatory afferents originating in the presubiculum. METHODS: Experiments were designed to ablate the presubiculum unilaterally by focal ibotenate injections and to evaluate the effect of this deafferentation on the vulnerability of EC layer III neurons to the chemoconvulsant kainate (injected systemically 5 days later). RESULTS: After treatment with kainate, 11 of the 15 rats preinjected with ibotenate showed clear cut, partial neuroprotection in layer III of the EC ipsilateral to the ibotenate lesion. Serial reconstruction of the ibotenate-induced primary lesion revealed that entorhinal neurons were protected only in animals that had lesions in the pre- and parasubiculum, especially in the deep layers (IV-VI). CONCLUSIONS: The deep layers of the pre- and parasubiculum appear to control the seizure-induced damage of EC layer III. This phenomenon may be of relevance for epileptogenesis and for the pathogenesis of temporal lobe epilepsy. PMID- 11488879 TI - Pharmacokinetics of levetiracetam and its enantiomer (R)-alpha-ethyl-2-oxo pyrrolidine acetamide in dogs. AB - PURPOSE: The new antiepileptic drug, levetiracetam (LEV, ucb LO59), is a chiral molecule with one asymmetric carbon atom whose anticonvulsant activity is highly enantioselective. The purpose of this study was to evaluate and compare the pharmacokinetics (PK) of LEV [(S)-alpha-ethyl-2-oxo-pyrrolidine acetamide] and its enantiomer (R)-alpha-ethyl-2-oxo-pyrrolidine acetamide (REV) after i.v. administration to dogs. This is the first time that the pharmacokinetics of both enantiomers has been evaluated. METHODS: Optically pure LEV and REV were synthesized, and 20 mg/kg of individual enantiomers was administered intravenously to six dogs. Plasma and urine samples were collected until 24 h, and the concentrations of LEV and REV were determined by an enantioselective assay. The levels of 2-pyrrolidone-N-butyric acid, an acid metabolite of LEV and REV, were determined by high-performance liquid chromatography (HPLC). The data were used for PK analysis of LEV and REV. RESULTS: LEV and REV had similar mean +/- SD values for clearance; 1.5 +/- 0.3 ml/min/kg and volume of distribution; 0.5 +/- 0.1 L/kg. The half-life (t1/2) and mean residence time (MRT) of REV (t1/2, 4.3 +/- 0.8 h, and MRT, 6.0 +/- 1.1 h) were, however, significantly longer than those of LEV (t1/2, 3.6 +/- 0.8 h, and MRT, 5.0 +/- 1.2 h). The renal clearance and fraction excreted unchanged for LEV and REV were significantly different. CONCLUSIONS: In addition to the enantioselective pharmacodynamics, alpha-ethyl-2-oxo-pyrrolidine acetamide has enantioselective PK. The enantioselectivity was observed in renal clearance. Because REV has more favorable PK in dogs than LEV, the higher antiepileptic potency of LEV is more likely due to intrinsic pharmacodynamic activity rather than to enantioselective PK. PMID- 11488880 TI - Anticonvulsant profile of valrocemide (TV1901): a new antiepileptic drug. AB - PURPOSE: We sought to investigate the anticonvulsant activity of the new antiepileptic drug (AED), valrocemide or TV1901 (VGD) in various animal (rodent) models of human epilepsy to determine its anticonvulsant profile and safety margin. METHODS: VGD was administered intraperitoneally to CF no. 1 mice and orally or intraperitoneally to Sprague-Dawley rats. The anticonvulsant activity of VGD was examined in nine different animal models of epilepsy for its ability to block electrically, chemically, or sensorily induced seizures. RESULTS: In mice VGD afforded complete protection against maximal electroshock (MES)-, pentylenetetrazole-, picrotoxin-, and bicuculline-induced seizures and 6-Hz "psychomotor" seizures with median effective dose (ED50) values of 151, 132, 275, 248, and 237 mg/kg, respectively. VGD was also effective in preventing sound induced seizures in Frings audiogenic-seizure susceptible mice (ED50, 52 mg/kg). The median neurotoxic dose in mice was 332 mg/kg. After oral administration to rats, VGD was active in the MES test, with an ED50 of 73 mg/kg, and the median neurotoxic dose was 1,000 mg/kg. Intraperitoneal administration of 300 mg/kg of VGD to hippocampal kindled Sprague-Dawley rats blocked generalized seizures and shortened the afterdischarge duration significantly. VGD also provided complete protection from focal seizures in the corneally kindled rats (ED50,161 mg/kg). CONCLUSIONS: The results obtained in this study suggest that VGD has a broad spectrum of anticonvulsant activity and promising potential as a new AED. PMID- 11488881 TI - Severe myoclonic epilepsy of infancy: extended spectrum of GEFS+? AB - PURPOSE: Severe myoclonic epilepsy of infancy (SMEI) is an intractable epilepsy of early childhood of unknown etiology. It is often associated with a family history of seizure disorders, but epilepsy phenotypes have not been well described. We sought to characterize the seizure phenotypes of relatives to better understand to the genetic basis of SMEI. METHODS: Probands with SMEI were identified, and systematic family studies were performed. Epilepsy syndromes were characterized in affected family members. RESULTS: Twelve probands with SMEI were identified. Eleven of the 12 probands with SMEI had a family history of seizures, and the twelfth was the result of a consanguineous marriage. We found that 16.7% of full siblings and 8.3% of parents had definite seizures. A total of 39 affected family members was identified. The most common phenotype was febrile seizures in 14, febrile seizures plus in seven, partial epilepsy in two, and there were single individuals with SMEI, myoclonic-astatic epilepsy, Lennox Gastaut syndrome, and 13 cases with unclassified or unconfirmed seizures. CONCLUSIONS: The family history of seizures in SMEI is in keeping with the spectrum of seizure phenotypes seen in generalized epilepsy with febrile seizures plus (GEFS+). Our findings suggest that SMEI is the most severe phenotype in the GEFS+ spectrum. PMID- 11488882 TI - Bilateral posterior parietal polymicrogyria: a mild form of congenital bilateral perisylvian syndrome? AB - PURPOSE: The main features of congenital bilateral perisylvian syndrome (CBPS) are pseudobulbar palsy, cognitive deficits, epilepsy, and perisylvian abnormalities on imaging studies, however, the clinical spectrum of this syndrome is much wider than previously believed and may vary from minor speech difficulties to severely disabled patients. The objective of this study was to present the different imaging and clinical findings of 17 patients with CBPS, their genetic background, and the occurrence of prenatal injury during their pregnancies. METHODS: We evaluated 17 consecutive patients with CBPS and divided them into two groups according to the imaging findings: (a) diffuse polymicrogyria around the sylvian fissure and (b) posterior polymicrogyria at the posterior parietal regions. They were systematically interviewed regarding history of prenatal events during their pregnancies, family history of speech difficulties, epilepsy, or other neurologic abnormality. RESULTS: There were seven women, ages ranging from 3 to 41 years (mean, 11.5; median, 7 years). Seven patients had bilateral posterior parietal polymicrogyria (BPPP), and 10 had diffuse bilateral perisylvian polymicrogyria. All seven patients with BPPP had only minor speech difficulties, none had epilepsy, and all but one had a family history of epilepsy or cortical dysgenesis. In contrast, 10 patients with diffuse bilateral perisylvian polymicrogyria had pseudobulbar palsy, four had epilepsy, eight had a history of a major prenatal event, and only four had a family history of epilepsy or developmental delay. CONCLUSIONS: These findings suggest that diffuse bilateral perisylvian polymicrogyria appears to be more related to injuries caused by environmental factors, whereas BPPP has a stronger genetic predisposition. In addition, BPPP appears to have a wider clinical spectrum than previously believed, and may represent a milder extreme within the spectrum of CBPS. PMID- 11488883 TI - Neuroimaging findings of cortical dyslamination with cytomegaly. AB - PURPOSE: Our aims were to identify (a) the characteristic magnetic resonance imaging (MRI) findings of cortical dyslamination with cytomegaly, including dysplastic and destructive lesions; (b) the relationship between MRI findings and pathologic characteristics; (c) the diagnostic role of functional neuroimaging studies in patients with these pathologies. METHODS: The series consisted of 23 adult patients who had proven cortical dyslamination with cytomegaly. The abnormalities found on MRI were subdivided according to the patterns of involvement. They also were compared with the patients' pathologic characteristics. With visual qualitative analysis, [18F]fluorodeoxyglucose with positron emission tomography (FDG-PET), and ictal single-photon emission computed tomography (SPECT), observations were classified as localizing, lateralizing, nonlateralizing, false-localizing, and false-lateralizing. The standard for correct localization of neuroimages was defined to be the resected lobe. RESULTS: Focal abnormalities were found in 14 cases by MRI. Six cases showed typical MRI findings of focal cortical dysplasia, with focal areas of cortical thickening with or without poor grey-white matter differentiation. Focal subcortical high signal intensities on T2-weighted images occurred in two cases. Six patients had the focal destructive pattern. Three of eight cases with normal MRI and four of eight cases with the nondestructive cortical dysplasia pattern had balloon cells. However, these were not found in six patients with the destructive MRI pattern. FDG-PET localized the pathologic lobe in 13 (65%) of 20 cases, and ictal SPECT achieved this in 11 (61.1%) of 18 cases. FDG-PET and ictal SPECT also correctly localized the pathologic lobe in four and two cases with normal MRI, respectively. CONCLUSIONS: Two distinct patterns of abnormal MRI were found in the pathology of cortical dyslamination with cytomegaly. These dysplastic and destructive patterns might reflect different pathogeneses, such as the time of insult. FDG-PET and ictal SPECT have confirmatory and independent diagnostic roles in localizing epileptogenic foci. PMID- 11488884 TI - The amygdala and temporal lobe simple partial seizures: a prospective and quantitative MRI study. AB - PURPOSE: To determine whether specific temporal lobe simple partial seizures (SPSs) are associated with an abnormal amygdala T2 (AT2) ipsilateral to the seizure focus in patients with intractable unilateral temporal lobe epilepsy (TLE). AT2 relaxation time mapping is a sensitive method for the detection of abnormal tissue in the amygdala in patients with refractory TLE. The relation between an abnormal AT2 in the epileptic temporal lobe and amygdala seizure onset has not been established. METHODS: Fifty patients with intractable unilateral TLE and concordant data during presurgical evaluation were included. Patients with a foreign-tissue lesion on standard magnetic resonance imaging (MRI) were excluded. All had AT2 mapping. Fifteen types of SPSs were ascertained prospectively, systematically, and blinded to the results of AT2 mapping. The SPSs of patients with a normal AT2 (n = 25) were compared with those of patients with an abnormal AT2 ipsilateral to the seizure focus (n = 25). RESULTS: The group of patients with an abnormal AT2 reported a median of six types of SPSs (range 1-11), in comparison with a median of three types of SPSs (range, 0-7) for the group with a normal AT2 (p<0.01). Deja vu, a warm sensation, an indescribable strange sensation, a cephalic sensation, and fear were associated with an abnormal AT2. The combination of deja vu, a cephalic sensation, a warm sensation, a gustatory hallucination, and an indescribable strange sensation discriminated best between the 25 patients with a normal and the 25 patients with an abnormal AT2. CONCLUSIONS: A high number and the types of different SPSs provide clinical evidence for early involvement of the amygdala during seizures in patients with refractory unilateral TLE and an abnormal AT2 in the epileptic temporal lobe PMID- 11488885 TI - The relationship between quantitative T2 relaxometry and memory in nonlesional temporal lobe epilepsy. AB - PURPOSE: We investigated the relationship between preoperative quantitative magnetic resonance imaging (MRI) T2 relaxometry and volumetry of the hippocampi and pre- and postoperative verbal memory in temporal lobectomy patients who had nonlesional temporal lobe epilepsy. METHODS: Pre- and postoperative memory data based on the Logical Memory (LM) subtest of the Wechsler Memory Scale-Revised (WMS-R) and the 30-min delayed recall trial of the Rey Auditory Verbal Learning Test (AVLT) were obtained from 26 left and 15 right temporal lobectomy patients. Coronal MRI T2 maps were generated for these 41 temporal lobectomy patients as well as 61 control patients. Hippocampal T2 relaxation times and hippocampal volumes, converted to z scores using control group data, were correlated with neuropsychological performance in the patients. RESULTS: In left temporal lobe onset patients, high T2 in the left hippocampal body predicted higher LM performance after surgery. Asymmetrically high T2 in the left hippocampal body (i.e., the right-minus-left difference), compared with the right hippocampal body, also predicted higher LM performance after surgery. In right temporal lobe onset patients, high T2 in the left hippocampal body predicted relatively lower AVLT performance after surgery. Multiple regression analysis in left temporal onset patients revealed that high T2 in the left hippocampal body together with higher preoperative LM performance predict higher postoperative LM performance. CONCLUSIONS: Our findings suggest that elevated (i.e., abnormal) hippocampal T2 signal is associated with memory ability (or hippocampal functional capacity) independent of MRI-determined hippocampal atrophy. Therefore, our findings support the use of quantitative T2 relaxometry as an independent predictor of verbal memory outcome in both left and right TLE patients who are candidates for temporal lobectomy. PMID- 11488886 TI - Interictal 99mTc-HMPAO SPECT in temporal lobe epilepsy: relation to clinical variables. AB - PURPOSE: Factors affecting blood flow observed by interictal single-photon emission computed tomography (SPECT) images in temporal lobe epilepsy (TLE) have not been systematically studied or consistently demonstrated. We evaluated interictal SPECT results with respect to many clinical variables in a large population of TLE patients, all of whom underwent temporal lobectomy. METHODS: Interictal 99mTc-HMPAO SPECT scans from 61 TLE patients were obtained before an anterior temporal lobectomy. SPECT was analyzed using a region of interest analysis (ROI) in the cerebellum, anterior temporal lobe, lateral temporal lobe, mesial temporal lobe, whole temporal lobe, and inferior frontal lobe. Asymmetry indices (AIs) were calculated. Correlative analysis of AIs and clinical variables was performed. RESULTS: The AIs from TLE patients differed significantly from those of controls in the anterior temporal (p < 0.01), lateral temporal (p < 0.001), and whole temporal (p < 0.01) regions. No consistent overall correlation between the AIs and clinical variables existed. In right TLE (RTLE) only, AIs in the lateral and whole temporal lobe were positively correlated with age of onset (r = 0.470, p < 0.05; r = 0.548, p < 0.01, respectively). Similarly, in RTLE only, duration of epilepsy was negatively correlated with the anterior (r = 0.395, p < 0.05) and mesial (r = -0.45, p < 0.05) temporal lobe AI. No correlations were found between clinical variables and AIs in left TLE (LTLE) patients. CONCLUSIONS: Significant correlation of age at onset and duration of epilepsy with AIs in RTLE but not LTLE suggests physiologic processes may be determined in part by laterality of TLE. Clinical applications are problematic. PMID- 11488887 TI - Interictal and ictal magnetoencephalographic study in patients with medial frontal lobe epilepsy. AB - PURPOSE: To determine whether magnetoencephalography (MEG) has any clinical value for the analysis of seizure discharges in patients with medial frontal lobe epilepsy (FLE). METHODS: Four patients were studied with 74-channel MEG. Interictal and ictal electroencephalographic (EEG) and MEG recordings were obtained. The equivalent current dipoles (ECDs) of the MEG spikes were calculated. RESULTS: In two patients with postural seizures, interictal EEG spikes occurred at Cz or Fz. The ECDs of interictal MEG spikes were localized around the supplementary motor area. In the other two patients with focal motor or oculomotor seizures, interictal EEG spikes occurred at Fz or Cz. The ECDs of interictal MEG spikes were localized at the top of the medial frontal region. The ECDs detected at MEG ictal onset were also localized in the same area as those of the interictal discharges. CONCLUSIONS: In medial FLE patients, interictal and ictal MEG indicated consistent ECD localization that corresponded to the semiology of clinical seizures. Our findings demonstrate that MEG is a useful tool for detecting epileptogenic focus. PMID- 11488888 TI - Medically intractable, localization-related epilepsy with normal MRI: presurgical evaluation and surgical outcome in 43 patients. AB - PURPOSE: High-resolution magnetic resonance imaging (MRI) plays a crucial role in the presurgical evaluation of patients with medically refractory partial epilepsy. Although MRI detects a morphologic abnormality as the cause of the epilepsy in the majority of patients, some patients have a normal MRI. This study was undertaken to explore the hypothesis that in patients with normal MRI, invasive monitoring can lead to localization of the seizure-onset zone and successful epilepsy surgery. METHODS: A series of 115 patients with partial epilepsy who had undergone intracranial electrode evaluation (subdural strip, subdural grid, and/or depth electrodes) between February 1992 and February 1999 was analyzed retrospectively. Of these, 43 patients (37%) had a normal MRI. RESULTS: Invasive monitoring detected a focal seizure onset in 25 (58%) patients, multifocal seizure origin in 12 (28%) patients, and in six patients, no focal seizure origin was found. Of the 25 patients with a focal seizure origin, cortical resection was performed in 24, of whom 20 (83%) had a good surgical outcome with respect to seizure control. Six of the 12 patients with multifocal seizure origin underwent other forms of epilepsy surgery (palliative cortical resection in two, anterior callosotomy in two, and vagal nerve stimulator placement in two). CONCLUSIONS: Successful epilepsy surgery is possible in patients with normal MRIs, but appropriate presurgical evaluations are necessary. In patients with evidence of multifocal seizure origin during noninvasive evaluation, invasive monitoring should generally be avoided. PMID- 11488889 TI - Results of surgery in patients with refractory extratemporal epilepsy with normal or nonlocalizing magnetic resonance findings investigated with subdural grids. AB - PURPOSE: To study the efficacy of extensive coverage of the brain surface with subdural grids in defining extratemporal cortical areas amenable for resection in patients with refractory extratemporal epilepy (R-ExTE) and normal or nonlocalizing magnetic resonance imaging (MRI) scans. METHODS: Sixteen patients with R-ExTE were studied. Eleven patients had simple partial, eight had complex partial, and three had supplementary motor area seizures. Seizure frequency ranged from three per month to daily episodes. Interictal EEG showed large focal spiking areas in 11 patients, secondary bilateral synchrony in four, and was normal in one patient. Surface ictal recordings were nonlocalizing in six patients, and in 10, they disclosed large ictal focal spiking areas. MRI was normal in 10 patients, and in six patients, focal nonlocalizing potentially epileptogenic lesions were found. All patients were given an extensive coverage of the cortical convexity with subdural electrodes through large unilateral (n = 13) or bilateral (n = 3) craniotomies. Bipolar cortical stimulation was carried out through the implanted electrodes. RESULTS: Interictal invasive recording findings showed widespread spiking areas in 13 patients and secondary bilateral synchrony in three. Ictal invasive recordings showed focal seizure onset in all patients. There were six frontal, two parietal, one temporooccipital, four rolandic, and three posterior quadrant resections. Thirteen patients had been rendered seizure free after surgery, and three had > or =90% of seizure-frequency reduction. Pathologic findings included gliosis (n = 10), cortical dysplasia (n = 5), or no abnormalities (n = 1). Six patients had transient postoperative neurologic morbidity. CONCLUSIONS: Extensive subdural electrodes coverage seems to be an effective way to investigate patients with R-ExTE and normal or nonlocalizing MRI findings. PMID- 11488890 TI - Spectrum of clinical and histopathologic responses to intracranial electrodes: from multifocal aseptic meningitis to multifocal hypersensitivity-type meningovasculitis. AB - PURPOSE: We sought to characterize and compare the histopathologic and clinical changes elicited by subdural and depth electrodes in subjects undergoing epilepsy surgery evaluation. METHODS: A retrospective review of clinical records, imaging and histopathologic studies of epilepsy surgery cases requiring subdural strips and depth electrodes for localization of epileptogenic tissue was performed between 1993 and 1999. Forty-nine subjects had a combination of subdural and depth, whereas 10 had depth electrodes only. Histopathologic changes were classified as mild, moderate, or severe based on the density, extent, and composition of the inflammatory infiltrate. RESULTS: Subdural electrodes induced a clinical picture of transient aseptic meningitis; histopathologically, the infiltrates were moderate in degree in the majority (73%) and severe in the remainder (27%), with T cells and eosinophils infiltrating the cortex and arteriolar walls (hypersensitivity-type response). Depth electrodes alone caused minimal or no symptoms of meningeal irritation; the cellular response elicited by these electrodes was mild in five and moderate in the remaining five cases; severe inflammation was not observed in this group. Although the proportion of small clinically silent hematomas was larger in cases with depth (five of 59) compared with subdural electrodes (one of 49), microhemorrhages were considerably more numerous with subdural than with depth electrodes. CONCLUSIONS: These results suggest that the spectrum of brain responses to foreign bodies is wide, ranging from self-limited physiologic to hypersensitivity-type reactions of varying severity. Subdural strips elicited more intense inflammation than did depth electrodes. The histopathologic extent of the reaction to either type of electrodes could not be precisely defined because of the retrospective nature of this study. History of allergy to latex or previous craniotomies are probable risk factors for the hypersensitivity-type reaction. Surgical outcome, excellent in the majority, was independent of the severity or type of inflammation, and there have not been neurologic or systemic sequelae. PMID- 11488891 TI - Material-specific memory changes after anterior temporal lobectomy as predicted by the intracarotid amobarbital test. AB - PURPOSE: The intracarotid amobarbital test (IAT) has been shown to predict verbal memory changes after anterior temporal lobectomy (ATL). Seeking to extend these findings, we examined two questions: (a) What is the relationship between material-specific aspects of IAT memory and material-specific memory changes after ATL? and (b) Which IAT memory score(s) optimally predict memory changes after surgery, the memory score after injection ipsilateral to the seizure focus, the memory score after injection contralateral to the seizure focus, or the IAT asymmetry score, comprising the ipsilateral minus contralateral injection scores? METHODS: Seventy left hemisphere language-dominant patients undergoing ATL for treatment of medically refractory seizures were administered a verbal and visuospatial recognition memory test before surgery and 3 weeks after surgery. IAT memory recognition scores for words and designs were used to predict verbal and visuospatial memory changes after surgery. RESULTS: After surgery, left ATL patients declined in verbal memory, whereas right ATL patients declined in visuospatial memory. IAT total recognition memory scores (collapsed across all types of materials) and IAT word memory scores were associated with postoperative verbal memory decline. This relationship was significant for the IAT ipsilateral injection memory scores and the IAT hemispheric asymmetry scores. IAT memory performances were not related to visuospatial memory changes. CONCLUSIONS: Results indicate IAT memory measures to be related to postoperative verbal, but not visuospatial, memory change. A specific relationship was found between postoperative verbal memory change and IAT verbal memory after injection ipsilateral to the seizure focus, when relying primarily on the contralateral hemisphere. This finding is consistent with the functional reserve model of memory change in ATL. PMID- 11488892 TI - Heart rate and heart rate variability changes in the intracarotid sodium amobarbital test. AB - PURPOSE: Changes in heart rate and heart rate variability have been found in prior studies performed during the intracarotid sodium amobarbital (ISA) test. However, these results are not entirely consistent with current models of differential cerebral involvement in the modulation of the heart. This study was designed to re-investigate this topic with a larger N than has heretofore been used. METHODS: The electrocardiogram was recorded during left and right ISAs in 73 subjects. Raw heart rate and heart rate variability were calculated. RESULTS: Raw heart rate increased during inactivation of either hemisphere, but more so for the right hemisphere. Heart rate variability changes consistent with decreasing parasympathetic tone also were found to occur during either ISA, but to a significant degree, only during right ISA. CONCLUSIONS: The right hemisphere appears to have a greater role in cerebral regulation of cardiac function, perhaps by virtue of the modification of parasympathetic effects. PMID- 11488893 TI - A study of the effect of color photostimulation from a cathode-ray tube (CRT) display on photosensitive patients: the effect of alternating red-cyan flicker stimulation. AB - PURPOSE: In an attempt to establish evidence for developing better guidelines for the production of animation programs that would not induce photosensitive seizures in Japan, we evaluated the effects of red flicker, alternating red/cyan (complementary color to red) flicker stimuli, and of contrast between the red and cyan frames from a cathode-ray tube (CRT) display in photosensitive patients. METHODS: We studied 35 photosensitive patients. They were exposed to seven types of flicker. The first three types were alternating red/cyan flicker (R/C) with the luminance of cyan set at three different levels, high, equal, and low luminance (65, 20, and 16 cd/m2, respectively) relative to the red (20 cd/m2). The following four types were red, cyan, yellow, and magenta flicker stimuli. EEGs were recorded while the patients watched these stimuli on a CRT display. RESULTS: Rates of photoparoxysmal response (PPR) provocation were 11.4, 13.7, and 14.0% with high-, no- and low-contrast R/C flicker, respectively, and 3.7% with red flicker. The differences between red and each of the other R/C flicker stimuli were all statistically significant (p<0.05, 0.01, 0.01). No significant differences were found between the effects by each of the three levels of contrast in alternating R/C flicker (p > 0.05). CONCLUSIONS: These findings suggest that alternating R/C flicker is more provocative than simple red flicker, and that contrast between frames of different colors may play some role in the effects of alternating flicker stimuli from a CRT display in photosensitive patients. Therefore, caution against the use of the combination of red and cyan, in addition to the red flicker stimulus, should be included in any guidelines drawn up to prevent photosensitive seizures. PMID- 11488894 TI - Thyroid function in men taking carbamazepine, oxcarbazepine, or valproate for epilepsy. AB - PURPOSE: Antiepileptic drugs (AEDs) may affect serum thyroid hormone concentrations. This study aimed to evaluate thyroid function in men taking carbamazepine (CBZ), oxcarbazepine (OCBZ), or valproate (VPA) for epilepsy. METHODS: Ninety men with epilepsy (40 taking CBZ, 29 taking OCBZ, and 21 taking VPA monotherapy) and 25 control subjects participated in the study. After clinical examination, a blood sample for hormone, gamma-glutamyl-transferase (GGT) and antibody (ab) assays was obtained. RESULTS: Serum thyroxine (T4) and free thyroxine (FT4) concentrations were low in men taking CBZ or OCBZ. Forty five percent of men taking CBZ and 24% of men taking OCBZ had serum T4 and/or FT4 levels below the reference range. However, no correlations were found between T4 or FT4 and GGT concentrations in men taking CBZ or OCBZ. Thirteen percent of men taking CBZ, 17% of men taking OCBZ, and 6% of control men had increased levels of thyroid peroxidase (TPO)-ab and/or thyroglobulin (TG)-ab, but these were not associated with altered serum thyroid hormone concentrations. Serum triiodothyronine and thyrotropin levels in men taking CBZ or OCBZ were normal. In men taking VPA, the concentrations of thyroid hormones, thyrotropin, and antithyroid ab were normal. CONCLUSIONS: Serum thyroid hormone concentrations are low in CBZ- or OCBZ-treated men. However, these low levels do not seem to be due to liver enzyme induction or activation of immunologic mechanisms. Therefore, interference with hypothalamic regulation of thyroid function by CBZ and OCBZ seems possible. VPA does not have any significant effects on thyroid function. PMID- 11488895 TI - Lamotrigine therapy of epilepsy in tuberous sclerosis. AB - PURPOSE: Lamotrigine (LTG), a newer antiepileptic drug (AED), has activity against both partial-onset and generalized seizures. Its reported benefits for behavior, and its effectiveness in Lennox-Gastaut syndrome and other forms of refractory epilepsy, make it a logical choice for treatment of epilepsy in tuberous sclerosis complex (TSC). We present our experience with LTG therapy of epilepsy in 57 patients with TSC. METHODS: Patients fulfilled the diagnostic criteria for clinically definite TSC. LTG was initiated and increased until improvement in seizure frequency was noted, intolerable side effects occurred, or maximal doses were reached. Seizure frequency and behavioral changes were recorded during LTG therapy and compared with those prior to the introduction of LTG. RESULTS: Twenty-four (42%) were seizure free, and 21 (37%) had a >50% reduction in seizure frequency. Eighteen (32%) had subjectively improved behavior and/or alertness with daily activities. Thirty-eight (67%) had no change in this regard, whereas one (2%) became worse. Responders were more likely to not have a history of infantile spasms, and to have experienced only partial seizures (p < 0.05). Otherwise no phenotypic correlations with response were apparent. CONCLUSIONS: Among patients with TSC and epilepsy, LTG was effective and well tolerated, including as initial monotherapy. Improved alertness and behavior were apparent in many patients. The incidence of side effects is similar to that reported for other pediatric populations with symptomatic partial epilepsy. The usefulness of LTG in TSC may relate to an underlying defect of glutamatergic neurotransmission in partial epilepsy. PMID- 11488896 TI - Absence seizures aggravated by valproic acid. AB - PURPOSE: To report on pediatric patients with absence epilepsy who experienced absence seizure aggravation while receiving valproic acid (VPA). METHODS: The charts of all children from four pediatric epilepsy clinics receiving VPA for absence epilepsy were reviewed. Patients were evaluated and followed up between 1994 and 2000. RESULTS: Eight cases (six boys) of absence seizure aggravation were detected. Mean age at seizure onset was 5.8 years (range, 3-12 years). Six patients had simple absence seizures, one had myoclonic absences, and one had absences with automatisms. The electroencephalogram in all cases depicted generalized 3-Hz spike-and-wave activities. All eight patients experienced an increase in the frequency of absence seizures within days of VPA introduction. Dose increments resulted in further seizure aggravation. Serum levels of VPA were within therapeutic range in all patients. No case was attributed to VPA-induced encephalopathy. All patients improved on VPA discontinuation. In five children, VPA was reintroduced, resulting in further seizure aggravation. CONCLUSIONS: VPA can occasionally provoke absence seizure aggravation in patients with absence epilepsy. PMID- 11488897 TI - Transient dystonias in three patients treated with tiagabine. AB - PURPOSE: Tiagabine (TGB) is a new antiepileptic drug (AED) with gamma aminobutyric acid (GABA)ergic mechanism of action. GABAergic compounds may influence the extrapyramidal system, probably via modulation of dopaminergic nigrostriatal neurons. A well-known side effect of TGB is probably dose-related extrapyramidal tremor. To our knowledge, acute dystonias associated with TGB treatment have yet to be described. METHODS: Three patients with transient acute dystonic reactions while taking TGB as add-on therapy with carbamazepine (CBZ) are presented. The focal limb dystonia in one case, an oromandibular dystonia in second, and writer's cramp in third one were observed. RESULTS: In all cases dystonic movements resolved spontaneously without discontinuation of TGB therapy and without any concomitant treatment. CONCLUSIONS: Tiagabine may cause various mild extrapyramidal side effects. All three cases reported were diagnosed with transient possibly drug-related dystonia after increase in TGB dose. It remain unclear whether dystonic movements are specific for patients treated with TGB/CBZ bitherapy. PMID- 11488898 TI - Fixation-off sensitivity in an adult with symptomatic occipital epilepsy. AB - An adult patient had EEG occipital spike fixation-off sensitivity and spontaneous occipital seizures due to perinatal asphyxia with bioccipital hemorrhage. EEG abnormalities consisted of repetitive posterior spikes that occurred when the eyes were closed and other conditions of fixation-off. Magnetic resonance imaging (MRI) documented severe posterior lesions with wallerian degeneration of the optic radiation. Electron capture detector single-photon emission computed tomography (ECD-SPECT) during the state of fixation-off showed regional hyperperfusion in the right posterior regions. We suggest that partial denervation of the primary visual cortex in this patient resulted in an increased cortical excitability and that the inhibitory effect of central vision and fixation leads to a suppression of spontaneous epileptic activity. This unusual symptomatic case may serve as a model for the pathophysiology in the more often reported cases of idiopathic benign epilepsy syndrome of childhood. PMID- 11488899 TI - A case of Ohtahara syndrome with olivary-dentate dysplasia and agenesis of mamillary bodies. AB - We report a patient with early infantile epileptic encephalopathy (EIEE) with suppression-burst (Ohtahara syndrome) associated with olivary-dentate dysplasia and agenesis of mamillary bodies is reported. Although those with Ohtahara syndrome are a heterogeneous group, virtually all reported cases are secondary to neuronal migrational disorders, sometimes only identified by detailed neuropathologic examination, as in this case report, which describes mamillary body agenesis as a not-yet-recognized anomaly associated with Ohtahara syndrome. All children with Ohtahara syndrome should have high-resolution magnetic resonance imaging (MRI) and detailed postmortem neuropathologic examinations. PMID- 11488900 TI - Vagus nerve stimulation for the treatment of bilateral independent temporal lobe epilepsy. AB - PURPOSE: We studied the effect of vagus nerve stimulation (VNS) on seizure reduction in patients with intractable epilepsy with bilateral independent temporal lobe foci. METHODS: Ten patients who met the criterion of the presence of two distinctive clinical and ictal EEG seizure patterns were identified and followed up for 1 year. RESULTS: Six patients had >50% reduction in their seizure frequency that persisted up to > or =1 year of follow-up, whereas four patients reported small or no reduction in their partial seizures. CONCLUSIONS: VNS is often effective and well tolerated in this select group of intractable epilepsy patients. PMID- 11488901 TI - Unsuspected atypical hemispheric dominance for language as determined by fMRI. AB - PURPOSE: We sought to illustrate the value of functional magnetic resonance imaging (fMRI) in the presurgical assessment of hemispheric dominance for language by means of an illustrative case report. METHODS: fMRI with two language paradigms was performed in a right-handed patient without familial sinistrality suffering from a left frontal focal epilepsy. RESULTS: Both fMRI paradigms revealed unequivocally lateralized right hemispheric activation. Atypical language representation was confirmed by Wada test. The further presurgical workup could be restricted to subdural strip recordings instead of the initially intended grid implantation. After resective surgery, no language deficits were apparent. CONCLUSIONS: Hemispheric dominance for language should be assessed by fMRI in all patients before surgery in areas potentially relevant for language in either cerebral hemisphere. fMRI may influence the further diagnostic workup and should be performed before other invasive diagnostic procedures. PMID- 11488902 TI - Corpora amylacea replace the hippocampal pyramidal cell layer in a patient with temporal lobe epilepsy. AB - A 39-year-old woman, with a 23-year history of medically intractable seizures of the complex partial type, underwent a left anterior temporal lobectomy and hippocampectomy. Histologic examinations revealed a massive occurrence of corpora amylacea, neuronal loss, and gliosis in the pyramidal cell layer of the hippocampus. The distribution of corpora amylacea in this case was quite distinctive and unlike the patterns of "nonspecific" corpora amylacea formation seen in aging or other neurodegenerative conditions. The pathogenetic aspects of the accumulation of these peculiar bodies are discussed in relation to hippocampal sclerosis. PMID- 11488903 TI - Metabolic conversion as a pre-receptor control mechanism for lipophilic hormones. AB - The majority of physiological effects mediated by steroids, retinoids and thyroids is accomplished by binding to members of the nuclear receptor superfamily of ligand activated transcription factors. The complex specific effects of lipid hormones depend not only on receptor expression, distribution and interactions, but also on the availability and metabolic conversion of the hormone itself. The cell-specific metabolic activation of inactive hormone precursors introduces a further level of hormonal regulation, and constitutes an important concept in endocrinology. The metabolic reactions carried out are achieved by dehydrogenases/reductases, hydroxylases and other enzymes, acting on ligands of the steroid/thyroid/retinoic hormone receptor superfamily. The concept implies that these tissue- and cell-specific metabolic conversions contribute to lipid hormone action, thus pointing to novel targets in drug development. All components of this signalling system, the hormone compounds, the receptor proteins, and modifying enzyme families originate from an early metazoan date, emphasizing the essential nature of all elements for development and diversification of vertebrate life. PMID- 11488904 TI - Spectroscopic characterization of metallothionein from the terrestrial snail, Helix pomatia. AB - The Cd-sequestering metallothionein (MT) isoform isolated from the midgut gland of Roman snails exposed to Cd supplements in the feed was characterized by compositional and spectroscopic analysis. The preparations contained nearly 5 mol of Cd, small amounts of Cu and about 1 mol of Zn per chain mass of 6620 Da, in numerical agreement with the apoprotein's measured capacity of firmly binding a maximum of 6 equivalents of Cd per molecule. As with other Cd-containing MTs the occurrence of a prominent Cd-mercaptide-specific shoulder at 250 nm in its absorption spectrum showed that Cd is complexed in tetrahedral symmetry by the cysteine residues of the protein, and the multiphasic ellipticity profile in the CD spectrum revealed that these complexes are joined to form one or more oligonuclear Cd-mercapto clusters. Both spectral features vanished with the removal of the metal but were reconstituted to maximum amplitudes by readdition of Cd to the metal-free apoprotein, provided precautions were taken to prevent air oxidation of the latter. Quantitative analysis of snail MT reconstituted with Cd established that the 18 cysteine side chains bind the metal in a 3-to-1 ratio; spectroscopic studies on fractionally restored forms demonstrated that the six Cd ions were bound to the apoprotein molecule in succession in two sets of three Cd ions each. Thus, one can infer from the observed stoichiometry and the coordinating preferences of Cd that this gastropod MT, like the Cd-bearing MTs of marine crustaceans, harboured the metal in two separate cyclically constructed Cd3Cys9 clusters. The snail clusters differed, however, from other MTs in their response to acidification. Their protolytic dissociation proceeded through two separate protonation steps with the manifestation of spectroscopically distinguishable intermediate forms. Thus, this snail isoform displays in its metal composition and its chemical and spectroscopic features both similarities and differences to other animal kingdom MTs. Its properties suggest that it serves an important role in the protection of the terrestrial gastropod from Cd. PMID- 11488905 TI - Galactofuranoic-oligomannose N-linked glycans of alpha-galactosidase A from Aspergillus niger. AB - Extracellular alpha-galactosidase A was purified from the culture filtrate of an over-producing strain of Aspergillus niger containing multiple copies of the encoding aglA gene under the control of the glucoamylase (glaA) promoter. Endoglycosidase digestion followed by SDS/PAGE, lectin and immunoblotting suggested that glycosylation accounted for approximately 25% of the molecular size of the purified protein. Monosaccharide analysis showed that this was composed of N-acetyl glucosamine, mannose and galactose. Mild acid hydrolysis, mild methanolysis, immunoblotting and exoglycosidase digestion indicated that the majority of the galactosyl component was in the furanoic conformation (beta-D galactofuranose, Galf). At least 20 different N-linked oligosaccharides were fractionated by high-pH anion-exchange chromatography following release from the polypeptide by peptide-N-glycosidase F. The structures of these were subsequently determined by fast atom bombardment mass spectrometry to be a linear series of Hex(7-26)HexHA(c2). Indicating that oligosaccharides from GlcNA(c2)Man(7), increasing in molecular size up to GlcNA(c2)Man(24) were present. Each of these were additionally substituted with up to three beta-Galf residues. Linkage analysis confirmed the presence of mild acid labile terminal hexofuranose residues. These results show that filamentous fungi are capable of producing a heterogeneous mixture of high molecular-size N-linked glycans substituted with galactofuranoic residues, on a secreted glycoprotein. PMID- 11488906 TI - Comparative analyses of the conformational stability of a hyperthermophilic protein and its mesophilic counterpart. AB - Comparison of the conformational stability of an O(6)-methylguanine-DNA methyltransferase (MGMT) from the hyperthermophilic archaeon Thermococcus kodakaraensis strain KOD1 (Tk-MGMT), and its mesophilic counterpart C-terminal Ada protein from Escherichia coli (Ec-AdaC) was performed in order to obtain information about the relationship between thermal stability and other factors, such as thermodynamic parameters, thermodynamic stability and other unfolding conditions. Tk-MGMT unfolded at Tm = 98.6 degrees C, which was 54.8 degrees C higher than the unfolding temperature of Ec-AdaC. The maximum free energy (DeltaG(max)) of the proteins were different; the value of Tk-MGMT (42.9 kJ.mol-1 at 29.5 degrees C) was 2.6 times higher than that of Ec-AdaC (16.6 kJ.mol-1 at 7.4 degrees C). The high conformational stability of Tk-MGMT was attributed to a 1.6-fold higher enthalpy value than that of Ec-AdaC. In addition, the DeltaG(max) temperature of Tk-MGMT was considerably higher (by 22.1 degrees C). The apparent heat capacity of denaturation (DeltaC(p)) of Tk-MGMT was 0.7-fold lower than that of Ec-AdaC. These three synergistic effects, increasing DeltaGmax, shifted DeltaG vs. temperature curve, and low DeltaC(p), give Tk-MGMT its thermal stability. Unfolding profiles of the two proteins, tested with four alcohols and three denaturants, showed that Tk-MGMT possessed higher stability than Ec-AdaC in all conditions studied. These results indicate that the high stability of Tk-MGMT gives resistance to chemical unfolding, in addition to thermal unfolding. PMID- 11488907 TI - Regulation of MDR1 promoter activity in human breast carcinoma cells by protein kinase C isozymes alpha and theta. AB - Increased levels of the protein kinase C (PKC) isoenzymes alpha and theta occur in conjunction with MDR1 gene expression in cells and tissues that have acquired a multidrug resistance (MDR) phenotype. Studies using PKC activators or antisense strategies against PKC suggest that activation of PKC engenders MDR1 gene transcription. In this study the potential roles of PKC-alpha and PKC-theta in MDR1 gene transcriptional regulation were explored. Human-derived MCF-7 breast cancer cells that lack constitutive expression of PKC-alpha or PKC-theta at detectable levels were transfected with full-length PKC-alpha or PKC-theta genes driven by the ecdysone promoter. Stable transfectants were selected by use of the appropriate antibiotics. Treatment of these cells with ponasterone A induced expression of PKC that was catalytically active and underwent translocation and down-regulation on exposure to 12-O-tetradecanoyl-13-phorbol acetate (TPA). These cells were used to analyse PKC-mediated regulation of the MDR1 promoter by further transient transfection with either 1073 bp of the MDR1 gene promoter or deletion fragments thereof to -8 bp, each linked to a chloramphenicol acetyl transferase (CAT) reporter gene. In PKC-alpha expressing cells TPA caused activation of all promoter fragments to -29 bp. This finding suggests that TPA inducible MDR1 transcription mediated through the TPA responsive factor early growth response 1 (EGR-1) in this region of the promoter may be due to activation of PKC-alpha. In contrast, PKC-theta activated only two MDR1 fragments, -982 and 612 bp. The effect of TPA on reporter gene expression was attenuated by the PKC inhibitor GF 109203X. These data suggest that MDR1 promoter transcription can be regulated by PKC-alpha and PKC-theta. The results support the search for therapeutic strategies directed specifically against PKC-alpha to ameliorate resistance of tumours against cytotoxic agents. PMID- 11488908 TI - Further evidence for the involvement of insulin receptor substrates in epidermal growth factor-induced activation of phosphatidylinositol 3-kinase. AB - In accordance with our recent results obtained with cultured rat hepatocytes [Fujioka, T. & Ui, M. (2001) Eur. J. Biochem. 268, 25-34], epidermal growth factor (EGF) gave rise to transient tyrosine phosphorylation of insulin receptor substrates (IRS-1 and IRS-2), thereby activating the bound phosphatidylinositol 3 kinase in human epidermoid carcinoma A431 cells normally abundant in EGF receptors (EGFR) and Chinese hamster ovary (CHO) cells transfected with full length EGFR. These actions of EGF, although much smaller in magnitude than those of insulin or IGF-I in the same cells, were accompanied by tyrosine phosphorylation of EGFR rather than insulin or IGF-I receptors, never observed in wild-type CHO cells expressing no EGFR, and totally inhibited by an inhibitor of EGFR kinase, AG1478, that was without effect on insulin or IGF-I actions. Recombinant IRS-1 was phosphorylated on tyrosines upon incubation with purified EGFR from A431 cells and 32P-labeled ATP. When CHO cells were transfected with C terminal truncated EGFR lacking three NPXY motifs responsible for direct binding to phosphotyrosine-binding domains of IRSs, no effect of EGF could be observed. We suggest that tyrosine phosphorylation of IRS-1 or IRS-2 could mediate EGFR induced activation of phosphatidylinositol 3-kinase in mammalian cells. PMID- 11488909 TI - Enzymatic and electrochemical oxidation of N-hydroxy compounds. Redox potential, electron-transfer kinetics, and radical stability. AB - A series of N-hydroxyacetanilide and 1-hydroxybenzotriazole analogues derivatized by various functional substituents were studied with regard to redox potential, oxidation by laccase, oxidative stability, and correlation to the electronic and steric properties of the substituents. It was found that substituents carrying conjugative/pi-electron function influenced the redox potential more than substituents carrying inductive/sigma-electron function, and that the electron transfer from an N-hydroxy moiety to laccase was significantly affected by the redox potential. Electron-withdrawing substituents tended to reduce the electron density on the N-hydroxy group, leading to higher redox potential and lower oxidation rate. Bulky substitution or absence of N-phenyl tended to increase the Km of the N-hydroxy group, leading to lower oxidation rate. Oxidized N-hydroxy compounds were stabilized by N-phenyl or N-carbonyl group, but not by N-azo or highly strained structure. Potential implication of these effects on laccase based, N-hydroxy compound-mediated biocatalysis is discussed. PMID- 11488910 TI - The effect of thiamine supplementation on tumour proliferation. A metabolic control analysis study. AB - Thiamine deficiency frequently occurs in patients with advanced cancer and therefore thiamine supplementation is used as nutritional support. Thiamine (vitamin B1) is metabolized to thiamine pyrophosphate, the cofactor of transketolase, which is involved in ribose synthesis, necessary for cell replication. Thus, it is important to determine whether the benefits of thiamine supplementation outweigh the risks of tumor proliferation. Using oxythiamine (an irreversible inhibitor of transketolase) and metabolic control analysis (MCA) methods, we measured an in vivo tumour growth control coefficient of 0.9 for the thiamine-transketolase complex in mice with Ehrlich's ascites tumour. Thus, transketolase enzyme and thiamine clearly determine cell proliferation in the Ehrlich's ascites tumour model. This high control coefficient allows us to predict that in advanced tumours, which are commonly thiamine deficient, supplementation of thiamine could significantly increase tumour growth through transketolase activation. The effect of thiamine supplementation on tumour proliferation was demonstrated by in vivo experiments in mice with the ascites tumour. Thiamine supplementation in doses between 12.5 and 250 times the recommended dietary allowance (RDA) for mice were administered starting on day four of tumour inoculation. We observed a high stimulatory effect on tumour growth of 164% compared to controls at a thiamine dose of 25 times the RDA. This growth stimulatory effect was predicted on the basis of correction of the pre existing level of thiamine deficiency (42%), as assayed by the cofactor/enzyme ratio. Interestingly, at very high overdoses of thiamine, approximately 2500 times the RDA, thiamine supplementation had the opposite effect and caused 10% inhibition of tumour growth. This effect was heightened, resulting in a 36% decrease, when thiamine supplementation was administered from the 7th day prior to tumour inoculation. Our results show that thiamine supplementation sufficient to correct existing thiamine deficiency stimulates tumour proliferation as predicted by MCA. The tumour inhibitory effect at high doses of thiamine is unexplained and merits further study. PMID- 11488911 TI - Thermodynamics of the folding of D-glyceraldehyde-3-phosphate dehydrogenase assisted by protein disulfide isomerase studied by microcalorimetry. AB - Thermodynamics of the refolding of denatured D-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) assisted by protein disulfide isomerase (PDI), a molecular chaperone, has been studied by isothermal microcalorimetry at different molar ratios of PDI/GAPDH and temperatures using two thermodynamic models proposed for chaperone-substrate binding and chaperone-assisted substrate folding, respectively. The binding of GAPDH folding intermediates to PDI is driven by a large favorable enthalpy decrease with a large unfavorable entropy reduction, and shows strong enthalpy-entropy compensation and weak temperature dependence of Gibbs free energy change. A large negative heat-capacity change of the binding, 156 kJ.mol(-1).K(-1), at all temperatures examined indicates that hydrophobic interaction is a major force for the binding. The binding stoichiometry shows one dimeric GAPDH intermediate per PDI monomer. The refolding of GAPDH assisted by PDI is a largely exothermic reaction at 15.0-25.0 degrees C. With increasing temperature from 15.0 to 37.0 degrees C, the PDI-assisted reactivation yield of denatured GAPDH upon dilution decreases. At 37.0 degrees C, the spontaneous reactivation, PDI-assisted reactivation and intrinsic molar enthalpy change during the PDI-assisted refolding of GAPDH are not detected. PMID- 11488912 TI - Binding interactions between barley thaumatin-like proteins and (1,3)-beta-D glucans. Kinetics, specificity, structural analysis and biological implications. AB - The specificity and kinetics of the interaction between the pathogenesis-related group of thaumatin-like proteins (PR5) in higher plants and (1,3)-beta-D-glucans have been investigated. Two thaumatin-like proteins with 60% amino-acid sequence identity were purified from extracts of germinated barley grain, and were designated HvPR5b and HvPR5c. Purified HvPR5c interacted with insoluble (1,3) beta-D-glucans, but not with cellulose, pustulan, xylan, chitin or a yeast mannoprotein. Tight binding was observed with unbranched and unsubstituted (1,3) beta-D-glucans, and weaker binding was seen if (1,6)-beta-linked branch points or beta-glucosyl substituents were present in the substrate. The HvPR5b protein interacted weakly with insoluble (1,3)-beta-D-glucans and did not bind to any of the other polysaccharides tested. This indicated that only specific barley PR5 isoforms interact tightly with (1,3)-beta-D-glucans. The complete primary structures of HvPR5b and HvPR5c were determined and used to construct molecular models of HvPR5b and HvPR5c, based on known three-dimensional structures of related thaumatin-like proteins. The models were examined for features that may be associated with (1,3)-beta-D-glucan binding, and a potential (1,3)-beta-D glucan-binding region was located on the surface of HvPR5c. No obvious structural features that would prevent binding of (1,3)-beta-D-glucan to HvPR5b were identified, but several of the amino acids in HvPR5c that are likely to interact with (1,3)-beta-D-glucans are not present in HvPR5b. PMID- 11488913 TI - Chicken interleukin-6. cDNA structure and biological properties. AB - Suppression subtractive hybridization technology was used to identify differentially expressed genes in spleens of chickens that had been treated with the synthetic immune modifier S-28463. One induced chicken gene encoded a protein with about 35% sequence identity to human interleukin-6 (IL-6). It consists of 241 amino acids including a putative N-terminal signal peptide of 47 residues. Bacterially expressed chicken IL-6 (ChIL-6) carrying a histidine tag in place of the signal peptide was biologically active: it induced proliferation of the IL-6 dependent murine hybridoma cell line 7TD1. The concentration of ChIL-6 required for half-maximal proliferative response was approximately 60 pg.mL-1. When injected intravenously into adult chickens, purified recombinant ChIL-6 induced an increase in serum corticosterone levels. Supernatants of chicken LMH and monkey COS-7 cells transiently transfected with a ChIL-6 expression construct induced proliferation of 7TD1 cells, demonstrating that recombinant ChIL-6 from eukaryotic cells is also active. PMID- 11488914 TI - The peroxidase activity of cytochrome c-550 from Paracoccus versutus. AB - Next to their natural electron transport capacities, c-type cytochromes possess low peroxidase and cytochrome P-450 activities in the presence of hydrogen peroxide. These catalytic properties, in combination with their structural robustness and covalently bound cofactor make cytochromes c potentially useful peroxidase mimics. This study reports on the peroxidase activity of cytochrome c 550 from Paracoccus versutus and the loss of this activity in presence of H2O2. The rate-determining step in the peroxidase reaction of cytochrome c-550 is the formation of a reactive intermediate, following binding of peroxide to the haem iron. The reaction rate is very low compared to horseradish peroxidase (approximately one millionth), because of the poor accessibility of the haem iron for H2O2, and the lack of a base catalyst such as the distal His of the peroxidases. This is corroborated by the linear dependence of the reaction rate on the peroxide concentration up to at least 1 M H2O2. Steady-state conversion of a reducing substrate, guaiacol, is preceded by an activation phase, which is ascribed to the build-up of amino-acid radicals on the protein. The inactivation kinetics in the absence of reducing substrate are mono-exponential and shown to be concurrent with haem degradation up to 25 mM H2O2 (pH 8.0). At still higher peroxide concentrations, inactivation kinetics are biphasic, as a result of a remarkable protective effect of H2O2, involving the formation of superoxide and ferrocytochrome c-550. PMID- 11488915 TI - Grass group I pollen allergens (beta-expansins) lack proteinase activity and do not cause wall loosening via proteolysis. AB - Group I grass pollen allergens make up a subgroup of the beta-expansin family of cell wall loosening proteins in plants. A recent study reported that recombinant Phl p 1, the group I allergen from timothy grass pollen, was associated with papain-like proteinase activity and suggested that expansins loosen the plant cell wall via proteolysis. We tested this idea with three experimental approaches. First, we evaluated three purified native group I allergens from timothy grass, ryegrass and maize (Phl p 1, Lol p 1, Zea m 1) using five proteinase assays with a variety of substrates. The proteins had substantial wall loosening activity, but no detectable proteolytic activity. Thus we cannot confirm proteolytic activity in the pollen allergen class of beta-expansins. Second, we tested the ability of proteinases to induce cell wall extension in vitro. Tests included cysteine proteinases, serine proteinases, aspartic proteinases, metallo proteinases, and aggressive proteinase mixtures, none of which induced wall extension in vitro. Thus, wall proteins are unlikely to be important load-bearing components of the plant cell wall. Third, we tested the sensitivity of beta-expansin activity and native wall extension activity to proteinase inhibitors. The results show that a wide range of proteinase inhibitors (phenylmethanesulfonyl fluoride, N-ethylmaleimide, iodoacetic acid, Pefabloc SC, and others) inhibited neither activity. From these three sets of results we conclude proteolysis is not a likely mechanism of plant cell wall loosening and that the pollen allergen class of beta-expansins do not loosen cell walls via a proteolytic mechanism. PMID- 11488916 TI - Probing the conformational state of a truncated staphylococcal nuclease R using time of flight mass spectrometry with limited proteolysis. AB - The conformational state of C-terminally truncated staphylococcal nuclease R (SNR135), with and without bound ligands, has been studied by performing limited proteolysis with a specific endoproteinase Glu-C followed by electrophoresis and mass spectrometry. Comparison of the accessibility of the cleavage sites shows that the C-terminal truncation of 14 amino-acid residues causes significant unfolding of the C-terminal part of alpha helix 1 and the center of alpha helix 2, but there is little effect on other regions of the nuclease, in particular the N-terminal subdomain, which includes the active site of the nuclease. The truncation also makes the overall conformation of the nuclease more loose and flexible. Binding of ligands makes helices 1 and 2 more resistant to protease Glu C attack and converts the partially unfolded state to a native-like state, although the conformational stability of the SNR135 complex is still much lower than that of the full-length enzyme. The results suggest that the amino-acid residues around the active site in the truncated nuclease are arranged in a similar topology to those in the full-length nuclease. The study shows that there is a clear-cut correlation between protease susceptibility and conformational stability of the protein, and the initial proteolytic events are the most critical for evaluating the conformational features of the protein. This study demonstrates how mass spectrometry can be combined with limited proteolysis to observe conformational changes induced by ligand binding. PMID- 11488917 TI - Interaction of hemoglobin with enterobacterial lipopolysaccharide and lipid A. Physicochemical characterization and biological activity. AB - The interaction of hemoglobin (Hb) with endotoxins [i.e. with enterobacterial deep rough mutant lipopolysaccharide (LPS) Re and the "endotoxic principle" of LPS, lipid A] was investigated using a variety of physical techniques and with two biological assays, tumor necrosis factor (TNF)-alpha induction in human mononuclear cells and the Limulus amebocyte lysate (LAL) assay. Fourier-transform IR-spectroscopic experiments indicate nonelectrostatic binding to the hydrophobic moiety with a slight rigidification of the lipid A acyl chains, and an increase in the inclination of the lipid A backbone with respect to the membrane surface from 35 degrees to more than 40 degrees due to Hb binding, but no change of the predominantly alpha-helical secondary structures of Hb due to LPS binding. From isothermal titration calorimetry, the molar [Hb] : [endotoxin] binding ratio lies between 1 : 3 and 1 : 5 molar. Synchrotron radiation X-ray diffraction measurements indicate a reorientation of the lipid A aggregates from one cubic structure to another, the final structure belonging to space group Q224. The LPS induced TNF-alpha production of mononuclear cells is enhanced by Hb, whereas in the LAL assay an LPS concentration-dependent increase or decrease was observed. Although a detailed mechanism of action cannot be given, the enhancement of LPS bioactivity can be understood in the light of the previously presented conformational concept; Hb induces an increase in the conical shape of the lipid A moiety of LPS, higher cross-section of the hydrophobic than the hydrophilic part, and of the inclination angle of the diglucosamine backbone with respect to the direction of the acyl chains. PMID- 11488918 TI - Characterization of novel structures of mannosylinositolphosphorylceramides from the yeast forms of Sporothrix schenckii. AB - Novel structures of glycoinositolphosphorylceramide (GIPC) from the infective yeast form of Sporothrix schenckii were determined by methylation analysis, mass spectrometry and NMR spectroscopy. The lipid portion was characterized as a ceramide composed of C-18 phytosphingosine N-acylated by either 2 hydroxylignoceric acid (80%), lignoceric (15%) or 2,3-dihydroxylignoceric acids (5%). The ceramide was linked through a phosphodiester to myo-inositol (Ins) which is substituted on position O-6 by an oligomannose chain. GIPC-derived Ins oligomannosides were liberated by ammonolysis and characterized as: Manpalpha1- >6Ins; Manpalpha1-->3Manpalpha1-->6Ins; Manpalpha1-->6Manpalpha1-->3Manpalpha1- >3Manpalpha1-->6Ins; Manpalpha1-->2Manpalpha1-->6Manpalpha1-->3Manpalpha1- >3Manpalpha1-->6Ins. These structures comprise a novel family of fungal GIPC, as they contain the Manpalpha1-->6Ins substructure, which has not previously been characterized unambigously, and may be acylated with a 2,3 dihydroxylignoceric fatty acid, a feature hitherto undescribed in fungal lipids. PMID- 11488919 TI - Highly efficient targeting and accumulation of a F(ab) fragment within the secretory pathway and apoplast of Arabidopsis thaliana. AB - To further improve antibody production in plants, constructs were designed to minimize transgene silencing and to retain a F(ab) fragment within the secretory pathway of transgenic Arabidopsis thaliana plants. The levels of antibody accumulation suggest that placing the sequences that encode Fd and light chain under the control of nonidentical 3' regions reduces susceptibility to post transcriptional gene silencing compared with when the individual polypeptide encoding sequences are placed under the control of identical 3' regions. High levels of accumulation (up to 6% of total soluble protein) were found for both secreted and intracellularly targeted antibody fragments. Immunofluorescence microscopic analysis showed that F(ab) fragments devoid of any additional C terminal sequence were efficiently secreted, whereas retention of F(ab) fragments within the endomembrane system of the secretory pathway was achieved by C terminal fusion of the DIKDEL sequence to the antibody light chain. Furthermore, analysis by immunoprecipitation and ELISA showed that intracellular retention of antibody fragments did not affect antigen-binding activity, and more than 80% of the isolated antibody fragments were found to bind antigen. Taken together, our results provide improvements to the technology of recombinant antibody production in transgenic plants. PMID- 11488920 TI - Identification of protein-coding genes in the genome of Vibrio cholerae with more than 98% accuracy using occurrence frequencies of single nucleotides. AB - The published sequence of the Vibrio cholerae genome indicates that, in addition to the genes that encode proteins of known and unknown function, there are 1577 ORFs identified as conserved hypothetical or hypothetical gene candidates. Because the annotation is not 100% accurate, it is not known which of the 1577 ORFs are true protein-coding genes. In this paper, an algorithm based on the Z curve method, with sensitivity, specificity and accuracy greater than 98%, is used to solve this problem. Twenty-fold cross-validation tests show that the accuracy of the algorithm is 98.8%. A detailed discussion of the mechanism of the algorithm is also presented. It was found that 172 of the 1577 ORFs are unlikely to be protein-coding genes. The number of protein-coding genes in the V. cholerae genome was re-estimated and found to be approximately 3716. This result should be of use in microarray analysis of gene expression in the genome, because the cost of preparing chips may be somewhat decreased. A computer program was written to calculate a coding score called VCZ for gene identification in the genome. Coding/noncoding is simply determined by VCZ > 0/VCZ < 0. The program is freely available on request for academic use. PMID- 11488921 TI - Recombinant human factor VIII-specific affinity ligands selected from phage displayed combinatorial libraries of protein A. AB - Factor VIII-specific affibodies were selected from phage displayed libraries constructed by combinatorial mutagenesis of an alpha helical bacterial receptor domain derived from staphylococcal protein A. Bead-immobilized recombinant human factor VIII (rVIII) (80 and 90 kDa chains) protein was used during competitive biopannings in the presence of free 80-kDa chain protein, resulting in the selection of several binders that showed dissociation constants (Kd) in the range 100-200 nM as determined by biosensor analyses. One variant (Z[rVIII:3], 90-kDa chain specific) was further characterized in small-scale affinity chromatography experiments, and showed efficient and selective recovery of biologically active rVIII from Chinese hamster ovary cell supernatant-derived feed stocks. The purity of the enriched rVIII was comparable with rVIII material purified by immunoaffinity chromatography using a 90-kDa chain-specific monoclonal antibody. Interestingly, epitope mapping showed that the monoclonal antibody and the affibody ligand competed for the same or at least overlapping epitopes on rVIII. In addition, the Z[rVIII:3] variant was produced by peptide synthesis with a C terminal cysteine to enable directed coupling to solid supports. This 59-residue protein was analyzed by circular dichroism and showed a secondary structure content similar to that of the parental Z domain used as scaffold. In biosensor studies, the synthetic affibody was immobilized recruiting the C-terminal cysteine residue, and demonstrated to bind both recombinantly produced and plasma derived factor VIII. From a secondary library, constructed by re-randomization of relevant positions identified after alignment of the first-generation variants, a panel of affinity-improved second-generation affibodies were selected of which one clone showed a dissociation constant (Kd) for rVIII of 5 nM. Several of these variants also showed higher apparent binding efficiencies towards rVIII when analyzed as immobilized ligands in biosensor experiments. Taken together, the results suggest that affibody ligands produced by bacterial or synthetic routes could be of interest as an alternative to monoclonal antibodies in purification processes or as diagnostic or monitoring tools. PMID- 11488922 TI - Crumpled structure of the custom hydrophobic lytic peptide cecropin B3. AB - The solution structure of a custom lytic peptide, cecropin B3 (CB3), having two identical hydrophobic segments on both the N- and C-termini, was investigated by two-dimensional NMR spectroscopy. The need to determine the structure of this peptide is rooted in its specific ability to lyse lipid layers that have a high content of anionic lipid. The lytic activities of CB3 on cell membranes including cancer cells and bacteria is found to be less than cecropin B1. The results show that CB3 has four discrete segments forming alpha helical structures. The crumpled structure of CB3 provides evidence for the lysis of the lipid layer being via a pathway that differs from pore formation. The results in this study provide strong clues towards a rational design for a potent antimicrobial and antitumor peptide. PMID- 11488923 TI - Estrogen induces a rapid secretion of amyloid beta precursor protein via the mitogen-activated protein kinase pathway. AB - The female sex hormone estrogen (17beta-estradiol; E2) may function as a neurohormone and has multiple neuromodulatory functions in the brain. Its potent neuroprotective activities can be dependent and independent of estrogen receptors (ERs). In addition, E2 influences the processing of the amyloid beta precursor protein (APP), one central step in the pathogenesis of Alzheimer's disease. Here, we show: (a) that physiological concentrations of E2 very rapidly cause an increased release of secreted nonamyloidogenic APP (sAPPalpha) in mouse hippocampal HT22 and human neuroblastoma SK-N-MC cells; and (b) that this effect is mediated through E2 via the phosphorylation of extracellular-regulated kinase 1 and 2 (ERK1/2), prominent members of the mitogen-activated protein kinase (MAPK) pathway. Furthermore, we show that the activation of MAPK-signaling pathway and the enhancement of the sAPP release is independent of ERs and could be induced by E2 to a similar extent in neuronal cells either lacking or overexpressing a functional ER. PMID- 11488924 TI - Phosphorylation of serine residues affects the conformation of the calmodulin binding domain of human protein 4.1. AB - We have previously characterized the calcium-dependent calmodulin (CaM)-binding domain (Ser76-Ser92) of the 135-kDa human protein 4.1 isoform using fluorescence spectroscopy and chemically synthesized nonphosphorylated or serine phosphorylated peptides [Leclerc, E. & Vetter, S. (1998) Eur. J. Biochem. 258, 567-671]. Here we demonstrate that phosphorylation of two serine residues within the 17-residue peptide alters their ability to adopt alpha helical conformation in a position-dependent manner. The helical content of the peptides was determined by CD-spectroscopy and found to increase from 36 to 45% for the Ser80 phosphorylated peptide and reduce to 28% for the Ser84 phosphorylated peptide; the di-phosphorylated peptide showed 32% helical content. Based on secondary structure prediction methods we propose that initial helix formation involves the central residues Leu82-Phe86. The ability of the peptides to adopt alpha helical conformations did not correlate with the observed binding affinities to CaM. We suggest that the reduced CaM-binding affinities observed for the phosphorylated peptides are more likely to be the result of unfavorable sterical and electrostatic interactions introduced into the CaM peptide-binding interface by the phosphate groups, rather than being due to the effect of phosphorylation on the secondary structure of the peptides. PMID- 11488925 TI - Novel hopanoids from Frankia spp. and related soil bacteria. Squalene cyclization and significance of geological biomarkers revisited. AB - Three series of hopanoids, differing by their configurations at C-17 and C-21, have been identified in several Frankia spp. and other related soil bacteria. The widespread bacterial hopanoids of the 17beta(H),21beta(H) series were accompanied by their isomers of the 17beta(H),21alpha(H) (moretane) and 17alpha(H), 21beta(H) series. The latter series has not previously been found in living organisms and is considered to be a result of the abiotic isomerization of the thermodynamically less stable 17beta(H),21beta(H) hopanoids. This simultaneous presence of three isomeric hopanoid series highlights intriguing problems in the biogenesis of the bacteriohopane skeleton and partly questions the significance of hopanic biomarkers in sediments. PMID- 11488926 TI - Thermodynamic analysis of the binding of glutathione to glutathione S-transferase over a range of temperatures. AB - The binding properties of a glutathione S-transferase (EC 2.5.1.18) from Schistosoma japonicum to substrate glutathione (GSH) has been investigated by intrinsic fluorescence and isothermal titration calorimetry (ITC) at pH 6.5 over a temperature range of 15-30 degrees C. Calorimetric measurements in various buffer systems with different ionization heats suggest that protons are released during the binding of GSH at pH 6.5. We have also studied the effect of pH on the thermodynamics of GSH-GST interaction. The behaviour shown at different pHs indicates that at least three groups must participate in the exchange of protons. Fluorimetric and calorimetric measurements indicate that GSH binds to two sites in the dimer of 26-kDa glutathione S-transferase from Schistosoma japonicum (SjGST). On the other hand, noncooperativity for substrate binding to SjGST was detected over a temperature range of 15-30 degrees C. Among thermodynamic parameters, whereas DeltaG degrees remains practically invariant as a function of temperature, DeltaH and DeltaS degrees both decrease with an increase in temperature. While the binding is enthalpically favorable at all temperatures studied, at temperatures below 25 degrees C, DeltaG degrees is also favoured by entropic contributions. As the temperature increases, the entropic contributions progressively decrease, attaining a value of zero at 24.3 degrees C, and then becoming unfavorable. During this transition, the enthalpic contributions become progressively favorable, resulting in an enthalpy-entropy compensation. The temperature dependence of the enthalpy change yields the heat capacity change (DeltaCp degrees ) of -0.238 +/- 0.04 kcal per K per mol of GSH bound. PMID- 11488927 TI - Domain structure of riboflavin synthase. AB - Riboflavin synthase of Escherichia coli is a homotrimer of 23.4 kDa subunits catalyzing the formation of the carbocyclic ring of the vitamin, riboflavin, by dismutation of 6,7-dimethyl-8-ribityllumazine. Intramolecular sequence similarity suggested that each subunit folds into two topologically similar domains. In order to test this hypothesis, sequence segments comprising amino-acid residues 1 97 or 101-213 were expressed in recombinant E. coli strains. The recombinant N terminal domain forms a homodimer that can bind riboflavin, 6,7-dimethyl-8 ribityllumazine and trifluoromethyl-substituted 8-ribityllumazine derivatives as shown by absorbance, circular dichroism, and NMR spectroscopy. Most notably, the recombinant domain dimer displays the same diastereoselectivity for ligands as the full length protein. The minimum N-terminal peptide segment required for ligand binding comprises amino-acid residues 1-87. The recombinant C-terminal domain comprising amino-acid residues 101-213 is relatively unstable and was shown not to bind riboflavin but to differentiate between certain diastereomeric trifluoromethyl-8-ribityllumazine derivatives. The data show that a single domain comprises the intact binding site for one substrate molecule. The enzyme catalyzed dismutation requires two substrate molecules to be bound in close proximity, and each active site of the enzyme appears to be located at the interface of an N-terminal and C-terminal domain. PMID- 11488928 TI - Stepwise proteolytic removal of the beta subdomain in alpha-lactalbumin. The protein remains folded and can form the molten globule in acid solution. AB - Bovine alpha-lactalbumin (alpha-LA) is an alpha/beta protein which adopts partly folded states when dissolved at low pH (A-state), by removal of the protein-bound calcium at neutral pH and low salt concentration (apo-state), as well as in aqueous trifluoroethanol. Previous spectroscopic studies have indicated that the A-state of alpha-LA at pH 2.0, considered a prototype molten globule, has a native-like fold in which the helical core is mostly retained, while the beta subdomain is less structured. Here, we investigate the conformational features of three derivatives of alpha-LA characterized by a single peptide bond fission or a deletion of 12 or 19/22 amino-acid residues of the beta subdomain of the native protein (approximately from residue 34 to 57). These alpha-LA derivatives were obtained by limited proteolysis of the protein in its partly folded state(s). A nicked alpha-LA species consisting of fragments 1-,3-40 and 41-123 (nicked-LA) was prepared by thermolytic digestion of the 123-residue chain of alpha-LA in 50% (v/v) aqueous trifluoroethanol. Two truncated or gapped protein species given by fragments 1-40 and 53-123 (desbeta1-LA) or fragments 1-34 and 54-,57-123 (desbeta2-LA) were obtained by digestion of alpha-LA with pepsin in acid or with proteinase K at neutral pH in its apo-state, respectively. The two protein fragments of nicked or gapped alpha-LA are covalently linked by the four disulfide bridges of the native protein. CD measurements revealed that, in aqueous solution at neutral pH and in the presence of calcium, the three protein species maintain the helical secondary structure of intact alpha-LA, while the tertiary structure is strongly affected by the proteolytic cleavages of the chain. Temperature effects of CD signals in the far- and near-UV region reveal a much more labile tertiary structure in the alpha-LA derivatives, while the secondary structure is mostly retained even upon heating. In acid solution at pH 2.0, the three alpha-LA variants adopt a conformational state essentially identical to the molten globule displayed by intact alpha-LA, as demonstrated by CD measurements. Moreover, they bind strongly the fluorescent dye 8 anilinonaphthalene-1-sulfonate, which is considered a diagnostic feature of the molten globule of proteins. Therefore, the beta subdomain can be removed from the alpha-LA molecule without impairing the capability of the rest of the chain to adopt a molten globule state. The results of this protein dissection study provide direct experimental evidence that in the alpha-LA molten globule only the alpha domain is structured. PMID- 11488929 TI - Cellulose-binding modules from extracellular matrix proteins of Dictyostelium discoideum stalk and sheath. AB - Cellulose-binding modules (CBMs) of two extracellular matrix proteins, St15 and ShD, from the slime mold Dictyostelium discoideum were expressed in Escherichia coli. The expressed proteins were purified to > 98% purity by extracting inclusion bodies at pH 11.5 and refolding proteins at pH 7.5. The two refolded CBMs bound tightly to amorphous phosphoric acid swollen cellulose (PASC), but had a low affinity toward xylan. Neither protein exhibited cellulase activity. St15, the stalk-specific protein, had fourfold higher binding affinity toward microcrystalline cellulose (Avicel) than the sheath-specific ShD CBM. St15 is unusual in that it consists of a solitary CBM homologous to family IIa CBMs. Sequence analysis of ShD reveals three putative domains containing: (a) a C terminal CBM homologous to family IIb CBMs; (b) a Pro/Thr-rich linker domain; and (c) a N-terminal Cys-rich domain. The biological functions and potential role of St15 and ShD in building extracellular matrices during D. discoideum development are discussed. PMID- 11488930 TI - Structure of the O-specific polysaccharide of Proteus mirabilis D52 and typing of this strain to Proteus serogroup O33. AB - The acidic O-specific polysaccharide chain (O-antigen) of the lipopolysaccharide (LPS) of Proteus mirabilis strain D52 was studied using chemical analyses along with 1H-NMR and 13C-NMR spectroscopy, including 2D COSY, TOCSY, ROESY, H-detected 1H,13C and 1H,31P HMQC experiments. The polysaccharide was found to contain D ribitol 5-phosphate (D-Rib-ol-5-P) and ethanolamine phosphate (Etn-P) and has the following structure: D-Rib-ol-5-P (3) approximately 75% EtnP(6)-->2)-beta-D-Galp (1-->3)-alpha-D-GlcpNAc-(1-->3)-beta-D-Glcp-(1-->3)-beta-D-GlcpNAc-(1-->). This structure is identical with that of the O-polysaccharide of P. mirabilis O33 strain 59/57, and, hence, P. mirabilis D52 belongs to the same Proteus serogroup O33. Serological studies with O-antiserum against P. mirabilis D52 confirmed this but showed that the LPS species of P. mirabilis 59/57 and D52 are not identical, having different epitopes in the core region. A serological cross-reactivity of P. mirabilis D52 O-antiserum was observed with LPS of two other Proteus strains, P. mirabilis O16 and P. penneri 103, which have structurally different O polysaccharides. The role of charged groups, Rib-ol-5-P and Etn-P in the immunospecificity is discussed. PMID- 11488931 TI - Improving solubility of catalytic domain of human beta-1,4-galactosyltransferase 1 through rationally designed amino acid replacements. AB - beta-1,4-galactosyltransferase 1 (beta4gal-T1, EC 2.4.1.38) transfers galactose from UDP-galactose to free N-acetyl-D-glucosamine or bound N-acetyl-D-glucosamine R. Soluble beta4gal-T1, purified from human milk has been refractory to structural studies by X-ray or NMR. In a previous study (Malissard et al. 1996, Eur. J. Biochem. 239, 340-348) we produced in the yeast Saccaromyces cerevisiae an N-deglycosylated form of soluble beta4gal-T1 that was much more homogeneous than the human enzyme, as it displayed only two isoforms when analysed by IEF as compared to 13 isoforms for the native beta4gal-T1. The propensity of recombinant beta4gal-T1 to aggregate at concentrations > 1 mg.mL(-1) prevented structural and biophysical studies. In an attempt to produce a beta4gal-T1 form suitable for structural studies, we combined site-directed mutagenesis and heterologous expression in Escherichia coli. We produced a mutated form of the catalytic domain of beta4gal-T1 (sfbeta4gal-T1mut) in which seven mutations were introduced at nonconserved sites (A155E, N160K, M163T, A168T, T242N, N255D and A259T). Sfbeta4gal-T1mut was shown to be much more soluble than beta4gal-T1 expressed in S. cerevisiae (8.5 mg.mL(-1) vs. 1 mg.mL(-1)). Catalytic activity and kinetic parameters of sfbeta4gal-T1mut produced in E. coli were shown not to differ to any significant extent from those of the native enzyme. PMID- 11488932 TI - Molecular characterization of Escherichia coli NAD kinase. AB - NAD kinase was purified to homogeneity from Escherichia coli MG1655. The enzyme was a hexamer consisting of 30 kDa subunits and utilized ATP or other nucleoside triphosphates as phosphoryl donors for the phosphorylation of NAD, most efficiently at pH 7.5 and 60 degrees C. The enzyme could not use inorganic polyphosphates as phosphoryl donors and was designated as ATP-NAD kinase. The N terminal amino-acid sequence of the purified enzyme was encoded by yfjB, which had been deposited as a gene of unknown function in the E. coli whole genomic DNA sequence database. yfjB was cloned and expressed in E. coli BL21(DE3)pLysS. The purified product (YfjB) showed NAD kinase activity, and was identical to ATP-NAD kinase purified from E. coli MG1655 in molecular structure and other enzymatic properties. The deduced amino-acid sequence of YfjB exhibited homology with that of Mycobacterium tuberculosis inorganic polyphosphate/ATP-NAD kinase [Kawai, S., Mori, S., Mukai, T., Suzuki, S., Hashimoto, W., Takeshi, Y. & Murata, K. (2000) Biochem. Biophys. Res. Commun. 276, 57-63], and those of many hypothetical proteins for which functions have not yet been revealed. The YfjB homologues were considered to be NAD kinases and alignment of their sequences revealed highly conserved regions, XXX-XGGDG-XL and DGXXX-TPTGSTAY, where X represents a hydrophobic amino-acid residue. PMID- 11488933 TI - Post-translational modification of the S-layer glycoprotein occurs following translocation across the plasma membrane of the haloarchaeon Haloferax volcanii. AB - The halophilic archaeon Haloferax volcanii is surrounded by a protein shell solely comprised of the S-layer glycoprotein. While the gene sequence and glycosylation pattern of the protein and indeed the three-dimensional structure of the surface layer formed by the protein have been described, little is known of the biosynthesis of the S-layer glycoprotein. In the following, pulse-chase radiolabeling and cell-fractionation studies were employed to reveal that newly synthesized S-layer glycoprotein undergoes a maturation step following translocation of the protein across the plasma membrane. The processing step, detected as an increase in the apparent molecular mass of the S-layer glycoprotein, is unaffected by inhibition of protein synthesis and is apparently unrelated to glycosylation of the protein. Maturation requires the presence of magnesium ions, involved in membrane association of the S-layer glycoprotein, and results in increased hydrophobicity of the protein as revealed by enhanced detergent binding. Thus, along with protein glycosylation, additional post translational modifications apparently occur on the external face of the haloarchaeal plasma membrane, the proposed topological homologue of the lumenal face of the eukaryal endoplasmic reticulum membrane. PMID- 11488934 TI - Effects of arsenic trioxide (As2O3) on leukemic cells from patients with non-M3 acute myelogenous leukemia: studies of cytotoxicity, apoptosis and the pattern of resistance. AB - Arsenic oxide (As2O3) has recently been reported to induce remission in a high percentage of patients with acute promyelocytic leukemia (APL). The aim of this study was to investigate the effects of As2O3 at therapeutic concentrations on cell viability and apoptosis on leukemic cells from patients with non-M3 acute myelogenous leukemia (AML) and to study the resistance profile compared to conventional AML drugs. Cells from 20 patients were exposed to therapeutic concentrations of As2O3 continuously for 96 h. As2O3 reduced the viability in blast cells from all the 20 tested patients compared to unexposed controls (p value: 0.02 at 0.05 microM; <0.005 at 1.0 microM and higher concentrations). An increase in the apoptotic rate was also seen after incubation with As2O3. Parallel to the incubation with arsenic the in vitro sensitivity to a number of chemotherapeutic agents commonly used in AML was studied. Correlation coefficients for the in vitro sensitivity were highly significant between the conventional AML drugs except for Ara-C. For As2O3, all the correlation coefficients were negative and ranged between -0.05 and -0.51. Furthermore, increased P-gp expression in a multidrug resistant HL-60 cell line did not decrease the sensitivity to As2O3 as compared to the parental cell line. Neither did a P-gp-transfected variant of the K562 cell line show decreased sensitivity to As2O3. We conclude that As2O3 at therapeutic concentrations induces apoptosis and cytotoxic effects in blast cells from patients with non-M3 AML, and that As2O3 differs from conventional AML drugs with respect to the mechanisms that confer resistance to the drugs. PMID- 11488935 TI - Platelet-derived growth factor (PDGF) in human acute myelogenous leukemia: PDGF receptor expression, endogenous PDGF release and responsiveness to exogenous PDGF isoforms by in vitro cultured acute myelogenous leukemia blasts. AB - We investigated effects of Platelet-derived growth factor (PDGF) and Platelet factor 4 (PF-4) on the functional characteristics of native, human acute myelogenous leukemia (AML) blasts. AML blast expression of the PDGF-receptor alpha-chain was detected for a subset of patients (45%), whereas PDGF-receptor beta-chain expression was detected for most patients (90%). Constitutive AML blast release of the PDGF-AB isoform (the major form also derived from normal platelets) was detected for 43% of patients, whereas PDGF-BB release was not detected for any patient. The PDGF isoforms AA, AB and BB had dose-dependent and divergent effects on spontaneous and cytokine-dependent AML blast proliferation, whereas for constitutive cytokine secretion (IL-1beta, IL-6, TNF-alpha) inhibitory effects were rare and all three isoforms usually had no effect or enhanced the constitutive secretion. The PDGF effects were caused by a direct effect on the AML blasts and were not dependent on the presence of serum. The PDGF effects could also be detected after in vitro culture of AML cells in the presence of IL-4+granulocyte-macrophage colony stimulating factor. PF-4 had divergent effects on proliferation and cytokine secretion by native AML blasts. Our results suggest that exogenous (e.g. platelet-secreted) PDGF and PF-4 can function as regulators of leukemic hematopoiesis and possibly also modulate the function of residual AML cells in peripheral blood stem cell grafts. On the other hand, endogenous release of PDGF-AB by native blasts may modulate the function of normal cells in the bone marrow microenvironment (e.g. bone marrow stromal cells). PMID- 11488936 TI - Modulation of CD44 in acute lymphoblastic leukemia identifies functional and phenotypic differences of human B cell precursors. AB - CD44 expression and other B cell markers were analyzed in 38 samples of B cell precursors (BCP) from patients with acute lymphoblastic leukemia (ALL). According to the expression of CD10 and CD44, we established the following five stages of BCP-ALL phenotypes that may represent different forms of interaction between BCP ALL and bone marrow-adherent cells: stage 1, CD19+, CD44bright, CD10-; stage 2, CD19+, CD44bright, CD10dim/bright; stage 3, CD19+, CD44dim, CD10bright, CD20-/+; stage 4, CD19+, CD44dim, CD10dim, CD20+; and stage 5, CD19+, CD44bright, CD10-, CD20+. Next, we analyzed the modulation of CD44 according to the expression of the different BCP-ALL phenotypes by incubating the samples under different culture conditions, including addition of stromal cells and interleukin (IL)-7. In culture, the samples in stages 1 and 2 maintained high expression of CD44 and re-expressed this molecule when cultured after trypsin treatment, indicating ongoing synthesis of CD44. Similarly, the stage 3 samples cultured in the presence of stromal cells, IL-7, or both also upregulated CD44 expression in culture. In contrast, the low expression of CD44 on the presumably more mature stage 4 samples was not modified by the addition of stromal cells or IL-7 or when cultured after trypsin treatment, suggesting that those cells had arrested CD44 synthesis. We concluded that down-modulation of CD44 occurred in association with differentiation to phenotype stages 3 and 4 and we hypothesized that this down modulation might be associated with the exit of BCP-ALL from the bone marrow. PMID- 11488937 TI - Increased risk for acute myeloid leukaemia in individuals with glutathione S transferase mu 1 (GSTM1) and theta 1 (GSTT1) gene defects. AB - OBJECTIVES: Glutathione S-transferases (GST) modulate the effects of exposure to various cytotoxic and genotoxic agents, including those associated with increased risks of the myelodysplastic syndrome (MDS), acute myeloid leukaemia (AML) and aplastic anemia (AA). Both the GST mu 1 (GSTM1) and GST theta 1 (GSTT1) genes have a null variant allele in which the entire gene is absent. In this study, we tested whether null genotypes for the GSTM1 and GSTT1 genes altered the risks for MDS, AML and AA. METHODS: Genomic DNA from 49 MDS, 38 AML and 37 AA patients and 276 controls was analysed using the polymerase chain reaction (PCR). RESULTS: The frequencies of GSTM1 (73.6%) and GSTT1 (34.2%) null genotypes were significantly higher in AML patients than in the controls (36.9 and 18.1%, respectively). A higher frequency of the combined null genotype for both genes was also observed in patients with AML (26.3% compared with 5.0% in the controls). In contrast, no differences in the frequencies of the null genotypes were found among MDS patients, AA patients and the controls. CONCLUSION: Our observation of a 4.7-fold (95% CI: 2.1-11.0) and 2.3-fold (95% CI: 1.0-5.2) increased risk associated with the GSTM1 and GSTT1 null genotypes, respectively, and a 6.6-fold (95% CI: 2.4 7.9) increased risk associated with the combined null genotype presents preliminary evidence that the inherited absence of this carcinogen detoxification pathway may be an important determinant of AML. PMID- 11488938 TI - Impact of vitamin A supplementation on anaemia and plasma erythropoietin concentrations in pregnant women: a controlled clinical trial. AB - INTRODUCTION: Although studies suggest that vitamin A or its metabolites influence the synthesis of erythropoietin in vitro and in animal models, it is unclear whether vitamin A supplementation increases plasma erythropoietin concentrations in humans. OBJECTIVE: To determine whether daily vitamin A supplementation increases plasma erythropoietin concentrations in pregnant women with a high prevalence of anaemia. METHODS: A randomized, double-blind, controlled clinical trial was conducted to examine the effect of daily vitamin A (3000 microg retinol equivalent), iron (30 mg), and folate (400 microg) versus iron (30 mg) and folate (400 microg) (control) on haemoglobin and plasma erythropoietin concentrations in 203 pregnant women in Malawi, Africa. RESULTS: Mean gestational age at enrollment was 23 wk, at which time 50% of the women were anaemic (haemoglobin <110 g/L). Mean (+/-SEM) change in haemoglobin from enrollment to 38 wk was 4.7+/-1.6 g/L (p=0.003) and 7.3+/-2.3 g/L (p=0.003) in the vitamin A and control groups, respectively. Mean change in plasma erythropoietin concentrations from enrollment to 38 wk was 2.39+/-5.00 (p=0.63) and -2.87+/-3.92 IU/L (p=0.46) in the vitamin A and controls groups, respectively. There were no significant differences between vitamin A and control groups in the slope of the regression line between log10 erythropoietin and haemoglobin at enrollment or 38 wk, and between enrollment and follow-up within either group. CONCLUSIONS: Vitamin A supplementation does not appear to increase haemoglobin and plasma erythropoietin concentrations among pregnant women with a high prevalence of anaemia in Malawi. PMID- 11488939 TI - Modulators of intraplatelet calcium concentration affect the binding of thrombospondin to blood platelets in healthy donors and patients with type 2 diabetes mellitus. AB - Thrombospondin (TSP), which is secreted from alpha-granules of activated platelets, binds to its surface receptor (CD36) in the presence of Ca2+. OBJECTIVES: We monitored how the modulation of intraplatelet Ca2+ affects TSP binding to CD36 on platelets from healthy donors and patients with type 2 diabetes mellitus. We also aimed to verify whether the impaired Ca2+ mobilisation in diabetes influences TSP binding upon the pharmacological modulation of calcium transport. METHODS: Whole blood cytometry was used to monitor TSP release/binding and CD36 presentation in platelets from 28 type 2 patients and 33 healthy donors. RESULTS: No significant changes in TSP and CD36 levels were revealed between the groups in circulating platelets and TRAP-, collagen- or thrombin-activated platelets. In healthy donors, 1 microM thapsigargin (TG) elevated the TRAP activated TSP binding (by up to 50%, p<0.001), 5 mM EGTA reversed the effect (by up to 85%, p<0.001), and overcame the effect of TG when used together. Less profoundly expressed effects occurred in the NIDDM group. In both groups TG increased the presentation of CD36 in TRAP-stimulated platelets (p<0.05), whereas EGTA lowered the TRAP-stimulated increase in CD36 (p<0.001). The inhibition of CD36 by EGTA was stronger in healthy volunteers (41% vs. 32%, respectively, p<0.05), whereas the activation by TG was higher in the NIDDM group (11% vs. 27%, p<0.05). When acting together the suppressive effects of EGTA on TG-dependent Ca2+ mobilisation were much attenuated in diabetic subjects (p<0.05). CONCLUSION: Both the release of TSP and CD36 presentation are under the influence of agents modulating intracellular Ca2+. Diabetic platelets seem more vulnerable to the releasers of cytosolic [Ca2+] and more resistant to the blockers of cytosolic [Ca2+] mobilisation. PMID- 11488940 TI - Acute tumour lysis syndrome following a single-dose corticosteroid in children with acute lymphoblastic leukaemia. AB - Acute tumour lysis syndrome (ATLS) is a well recognised complication of treatment of a variety of malignant disorders. It commonly occurs in patients with non Hodgkin's lymphoma (NHL) and acute lymphoblastic leukaemia (ALL) with the administration of combined cytotoxic chemotherapy. It is rarely reported after single-agent corticosteroid therapy. We present two children with acute lymphoblastic leukaemia of T-cell lineage who developed acute tumour lysis syndrome after a single dose of prednisolone, and methylprednisolone at the beginning of the induction chemotherapy. In the first case (an 11-yr-old) ATLS had occurred after an oral dose of prednisolone as small as 12 mg and within 18 h. The second case was a 14-yr-old boy with ALL who developed ATLS following a single dose of methylprednisolone. A few similar cases in the English literature are summarised in the report. These cases indicate that acute tumour lysis syndrome may occur after a single dose of corticosteroids. One should be aware of this potentially life-threatening complication especially while prescribing corticosteroids to patients with NHL and leukaemia. PMID- 11488941 TI - Sustained remission of platelet counts following monoclonal anti-CD20 antibody therapy in two cases of idiopathic autoimmune thrombocytopenia and neutropenia. AB - Idiopathic autoimmune thrombocytopenia and neutropenia (ITN) is a primary haemocytopenic disorder clinically characterised by recurrent mucocutaneous bleeding episodes and infections. Unlike in simple idiopathic thrombocytopenic purpura, the platelet deficiency of ITN tends to be chronic and difficult to treat. We describe two patients with ITN who obtained sustained remission of their platelet counts after therapy with the chimeric monoclonal anti-CD20 antibody Rituximab. In one of two cases, Rituximab also induced prolonged normalisation of the neutrophil count and disappearance of auto-antibodies. Our observations indicate that disturbed B-cell function plays a central role in the pathogenesis of ITN. Anti-CD20 antibody therapy seems to constitute a safe and efficient alternative to corticosteroids for the management of ITN patients with chronic thrombocytopenia. PMID- 11488942 TI - A novel stomatocytosis variant showing marked abnormalities in intracellular [Na] and [K] with minimal haemolysis. PMID- 11488943 TI - Analysis of cell signalling in the rodent pineal gland deciphers regulators of dynamic transcription in neural/endocrine cells. AB - In neurons, a temporally restricted expression of cAMP-inducible genes is part of many developmental and adaptive processes. To understand such dynamics, the neuroendocrine rodent pineal gland provides an excellent model system as it has a clearly defined input, the neurotransmitter norepinephrine, and a measurable output, the hormone melatonin. In this system, a regulatory scenario has been deciphered, wherein cAMP-inducible genes are rapidly activated via the transcription factor phosphoCREB to induce transcriptional events necessary for an increase in hormone synthesis. However, among the activated genes is also the inhibitory transcription factor ICER. The increasing amount in ICER protein leads ultimately to the termination of mRNA accumulation of cAMP-inducible genes, including the gene for the Aa-nat that controls melatonin production. This shift in ratio of phosphoCREB and ICER levels that depends on the duration of stimulation can be interpreted as a self-restriction of cellular responses in neurons and has also been demonstrated to interfere with cellular plasticity in many non-neuronal systems. PMID- 11488944 TI - N-methyl-D-aspartate receptor-mediated increase of neurogenesis in adult rat dentate gyrus following stroke. AB - Neurogenesis in the adult rat dentate gyrus was studied following focal ischemic insults produced by middle cerebral artery occlusion (MCAO). Animals were subjected to either 30 min of MCAO, which causes damage confined to the striatum, or 2 h of MCAO, which leads to both striatal and cortical infarction. When compared to sham-operated rats, MCAO-rats showed a marked increase of the number of cells double-labelled for 5-bromo-2'-deoxyuridine-5'-monophosphate (BrdU; injected during 4-6 days postischemia) and neuronal-specific antigen (NeuN; a marker of postmitotic neurons) in the ipsilateral dentate granule cell layer and subgranular zone at 5 weeks following the 2 h insult. Only a modest and variable increase of BrdU-labelled cells was found after 30 min of MCAO. The enhanced neurogenesis was not dependent on cell death in the hippocampus, and its magnitude was not correlated to the degree of cortical damage. Systemic administration of the N-methyl-D-aspartate (NMDA) receptor blocker dizocilpine maleate (MK-801) completely suppressed the elevated neurogenesis following 2 h of MCAO. Our findings indicate that stroke leads to increased neurogenesis in the adult rat dentate gyrus through glutamatergic mechanisms acting on NMDA receptors. This modulatory effect may be mediated through changes in the levels of several growth factors, which occur after stroke, and could influence various regulatory steps of neurogenesis. PMID- 11488945 TI - Calcium-activated potassium current clamps the dark potential of vertebrate rods. AB - Vertebrate photoreceptors respond to light with a graded hyperpolarization from a membrane potential in the dark of approximately -35 mV. The present work investigates the physiological role of the Ca2+-activated K+ current in the photovoltage generation in mechanically isolated rods from salamander retina. Membrane current or voltage in isolated rods was recorded from light- and dark adapted rods under voltage- or current-clamp conditions, respectively. In light adapted rods of the salamander, selective blockade of Ca2+-activated K+ channels by means of charybdotoxin depolarized the plasma membrane of current-clamped rods by approximately 30 mV, from a resting potential of approximately -35 mV. A similar depolarization was observed if external Ca2+ (1 mM) was substituted with Ba2+ or Sr2+. Under control conditions, the injection of currents of increasing amplitude (up to -100 pA, to mimic the current entering the rod outer segment) could not depolarize the membrane potential beyond a saturating value of approximately -20 mV. However, in the presence of charybdotoxin, rods depolarized up to +20 mV. In experiments with dark-adapted current-clamped rods, charybdotoxin perfusion lead to transient depolarizations up to 0 mV and steady state depolarizations of approximately 5 mV above the dark resting potential. Finally, the recovery phase of the voltage response to a flash of light in the presence of charybdotoxin showed a transient overshoot of the membrane potential. It was concluded that Ca2+-activated K+ current is necessary for clamping the rod photovoltage to values close to the dark potential, thus allowing faithful single photon detection and correct synaptic transmission. PMID- 11488946 TI - Chronic levodopa therapy does not improve skilled reach accuracy or reach range on a pasta matrix reaching task in 6-OHDA dopamine-depleted (hemi-Parkinson analogue) rats. AB - L-dopa therapy reverses some but not all of the motor deficits in human Parkinson patients. Although a number rat analogues of human Parkinson's disease have been developed for evaluating the efficacy of drug therapies, it is not known whether L-dopa has a similar selective action on the motor symptoms in the rat models. To examine the effectiveness of L-dopa in reversing the motor deficits in rats, we administered 6-OHDA unilaterally to produce hemi-Parkinson rats, which were then trained to reach for food using either their impaired (contralateral to the lesion) limb or their good (ipsilateral to the lesion) limb. To assess the skill, accuracy and range of limb movement, rats reached for pasta from a horizontal array of 260 vertically orientated pieces of pasta. The number and location of pasta pieces taken from this matrix was calculated and the qualitative aspects of the reaching movements were rated. The quantitative data on pasta sticks retrieved indicated that forelimb extension and movement radius around the shoulder joint was reduced by 6-OHDA treatment and did not improve after chronic L-dopa treatment. The qualitative analysis showed that grasping patterns, paw movements and body movements impaired by the lesion were also not improved by L dopa treatment. These findings are the first in the rat to suggest that whereas L dopa has a general activating effect on the rat's whole-body movements, as displayed in contralateral rotation, its effectiveness does not extend to skilled forelimb movements. The results are discussed in relationship to the idea that the restoration of some skilled movements may require normal synaptic function. PMID- 11488947 TI - Inhibition of synaptically evoked cortical acetylcholine release by intracortical glutamate: involvement of GABAergic neurons. AB - Cortical acetylcholine (ACh) has been shown to regulate diverse cognitive processes and its release can be regulated by neuromodulators that act presynaptically at cholinergic terminals. The neocortex receives dense glutamatergic input from thalamocortical and other fibres. The present study used in vivo microdialysis to examine, and pharmacologically characterize, the effect of glutamate on cortical ACh release evoked by electrical stimulation of the pedunculopontine tegmental nucleus in urethane-anaesthetized rats. All drugs were administered locally within the cortex by reverse dialysis. Application of glutamate had no detectable effect on spontaneous ACh release but reduced evoked cortical ACh efflux in a concentration-dependent manner. This effect was mimicked by the glutamate transporter blocker L-trans-pyrrolidine-2,4-dicarboxylic acid, as well as by the ionotropic glutamate receptor agonists N-methyl-D-aspartic acid and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, and was blocked by the ionotropic glutamate receptor antagonists 6,7-dinitroquinoxaline-2,3-dione and (+/-)-3-(2-carboxypiperazin-4yl)-propyl-1-phosphonic acid. Glutamate application also increased extracellular adenosine levels but the simultaneous delivery of the broad-spectrum adenosine receptor antagonist caffeine failed to affect the inhibitory action of glutamate on evoked ACh release. However, the effect of glutamate was fully blocked by simultaneous delivery of the GABAA receptor antagonist bicuculline and partially blocked by the GABAB receptor antagonist phaclofen. These results suggest that ionotropic glutamate receptor activation by glutamate inhibits evoked cortical ACh release via an indirect pathway involving GABAergic neurons in the cortex. PMID- 11488948 TI - Cellular and subcellular morphological localization of normal prion protein in rodent cerebellum. AB - Normal cellular prion protein, a necessary protagonist in fatal neurodegenerative prion diseases, was mapped in rodent cerebellum to establish its cellular and ultrastuctural localization. Existing morphological data about native prion protein distribution in brain tissues remain, indeed, contradictory and do not fit with biochemical and cell biological results. Using ultrastructural preembedding immunocytochemistry and a monoclonal anti-mouse prion protein antibody, this report shows that cellular prion protein is present in all cortico cerebellar and deep nuclei neuronal cell types, as well as in all glial cell types. The heaviest expression appears on parallel fibres and astrocytic processes. The protein is exclusively located on the outer cell membrane and in Golgi and endosomal intracytoplasmic organelles, with no cytoplasmic or synaptic vesicle labelling. Most important, and in contrast with previous ultrastructural data, cellular prion protein is shown to be distributed on all portions of neurons, without any preferential synaptic targeting. The present morphological report shows, for the first time in vivo, that the cellular prion protein is present on the entire cell surface membrane of all neuronal and glial cell types of the rat cerebellum. This ubiquitous presence supports the notion that prion protein has a generalized cellular function in brain tissue rather than a specialized role restricted to synaptic transmission. PMID- 11488949 TI - Purification of Purkinje cells by fluorescence-activated cell sorting from transgenic mice that express green fluorescent protein. AB - The cerebellar Purkinje cell has been the focus of numerous studies involving the analysis of development and information processing in the nervous system. Purkinje cells represent less than 0.1% of the total cell content of the cerebellum. To facilitate studies of molecules that are expressed in such a small proportion of neurons, we have established procedures for the purification of these cells. Transgenic mice were developed in which the expression of green fluorescent protein (GFP) was controlled by the L7 promoter. In adult cerebellum, GFP fluorescence was only detected in Purkinje cells, where it filled dendrites, soma and axons. GFP fluorescence was detected in Purkinje cells as early as embryonic day 17 and increased during development in vivo and in dissociated cerebellar culture. Mirroring endogenous L7 expression, high levels of GFP were observed in retinal rod bipolar cells. Lower levels of GFP were seen in olfactory periglomerular cells, neurons in the interpeduncular nucleus, and superior colliculus neurons. Cerebella from transgenic mice were dissociated by mild enzymatic treatment and Purkinje cells were isolated by fluorescence-activated cell sorting (FACS). By selecting optimal parameters, a fraction of viable Purkinje cells that was 94% pure was obtained. These results indicate that FACS is a powerful tool for isolating Purkinje cells from postnatal L7-GFP transgenic mice. GFP-positive neurons will also be useful in the real-time observation of dendritic morphogenesis and axonal outgrowth during development, or after neuronal activity in vitro. PMID- 11488950 TI - Expression and regulation of leptin receptor proteins in afferent and efferent neurons of the vagus nerve. AB - Leptin, the product of the ob gene, plays a key role in the regulation of food intake via a cross-talk between hypothalamic leptin receptors and neuropeptides that affect feeding behaviour. Recent studies have shown a synergistic interaction between leptin and cholecystokinin (CCK) leading to suppression of food intake, which involves CCK-1 receptors and capsaicin-sensitive vagal fibres. In this study, we have investigated the presence of leptin receptors in afferent and efferent neurons of the vagus nerve. By using reverse transcription polymerase chain reaction, mRNAs encoding long (Ob-Rb) and short (Ob-Ra) leptin receptor isoforms were detected in the rat nodose ganglion, which contains the cell bodies of the vagal afferent neurons. Western blot analysis confirmed the presence of leptin receptor-immunoreactive proteins in extracts from the vagal trunk. Immunohistochemistry showed the presence of leptin receptors and the leptin-induced transcription factor STAT3 in the cytoplasm of nodose ganglion cells. In cervical vagal segments, levels of leptin receptor protein displayed physiological regulation, with decreased amounts after feeding and increased levels after food restriction. In addition, leptin receptor and STAT3 immunoreactivities were detected in neurons of the nucleus of tractus solitarius (NTS) and the dorsal motor nucleus of the vagus nerve (DMNX) by immunofluorescence histochemistry. Furthermore, direct double-labelling demonstrated colocalization of Ob-Rb and STAT3 immunoreactivities in cholinergic vagal efferent cell bodies of the DMNX. It is speculated that vagal leptin receptors, apart from being activated by adipocyte-derived leptin, may also be influenced by leptin produced by the stomach. This may explain the synergistic action of leptin and CCK on neuronal activity in the NTS and on food intake. PMID- 11488951 TI - Ca2+-dependent regulation of synaptic delta2 glutamate receptor density in cultured rat Purkinje neurons. AB - The delta2 glutamate receptor (delta2 receptor), which is expressed abundantly at parallel fibre-Purkinje neuron synapses, has an important role in synaptogenesis and synaptic plasticity in the cerebellum. The present study examined the molecular mechanisms regulating synaptic delta2 receptor density. Immunocytochemistry, using two antibodies against the intracellular C-terminal and extracellular N-terminal regions of the delta2 receptor, indicated the reversible redistribution of postsynaptic delta2 receptors in response to either glutamatergic stimulation or enhancement of synaptic activity. The effect of glutamatergic stimulation was completely inhibited by either coapplication of the glutamate receptor antagonist or the removal of extracellular Ca2+ using EGTA and mimicked by selective activation of voltage-gated Ca2+ channels (VGCCs) with KCl, suggesting the significant role of Ca2+ influx in delta2 receptor redistribution. Biochemical examination indicated that a large amount of delta2 receptor protein was internalized following glutamatergic stimulation. These results suggest that the number of synaptic delta2 receptors is controlled by endocytosis in a synaptic activity- and intracellular Ca2+-dependent way, through which synaptogenesis and synaptic plasticity in Purkinje cells might be modulated. PMID- 11488952 TI - Fluorescent tracer in pilocarpine-treated rats shows widespread aberrant hippocampal neuronal connectivity. AB - Neuronal fibres of the hippocampal formation of normal and chronic epileptic rats were investigated by fluorescent tracing methods using the pilocarpine model of limbic epilepsy. Two months after onset of spontaneous limbic seizures, hippocampal slices were prepared and maintained in vitro for 10 h. Small crystals of fluorescent dye [fluorescein (fluoro-emerald) and tetramethylrhodamine (fluoro ruby)] were applied to different hippocampal regions. The main findings were: (i) in control rats there was no supragranular labelling when the mossy fibre tract was stained in stratum radiatum of area CA3. However, in epileptic rats a fibre network in the inner molecular layer of the dentate gyrus was retrogradely labelled; (ii) a retrograde innervation of area CA3 by CA1 pyramidal cells was disclosed by labelling remote CA1 neurons after dye injection into the stratum radiatum of area CA3 in chronic epileptic rats; (iii) labelling of CA1 neurons apart from the injection site within area CA1 was observed in epileptic rats but not in control animals; and (iv), a subicular-hippocampal projection was present in pilocarpine-treated rats when the tracer was injected just below the stratum pyramidale of area CA1. The findings show that fibre rearrangement in distinct regions of the epileptic hippocampal formation can occur as an aftermath of pilocarpine-induced status epilepticus. PMID- 11488953 TI - Effects of cannabinoids on prefrontal neuronal responses to ventral tegmental area stimulation. AB - Cannabinoids activate the firing of mesoprefrontocortical dopamine neurons and release dopamine in the prefrontal cortex. This study was undertaken with the aim of clarifying the interaction between cannabinoids and mesocortical system in the prefrontal cortex. The effect of Delta9-tetrahydrocannabinol (Delta9-THC) and the synthetic CB1 agonist WIN55,212-2 (WIN) was studied by extracellular single unit recordings, in chloral hydrate anaesthetised rats, on the spontaneous activity of pyramidal neurons and on the inhibition produced on these neurons by the electrical stimulation of the ventral tegmental area (VTA). Intravenously administered Delta9-THC and WIN (1.0 and 0.5 mg/kg, respectively), increased the firing rate of pyramidal neurons projecting to the VTA. VTA stimulation produced a phasic inhibition (167 +/- 6 ms) in 79% of prefrontal cortex pyramidal neurons. Delta9-THC and WIN reverted this inhibition in 73% and 100% of the neurons tested, respectively. The subsequent administration of the selective CB1 antagonist SR141716A (1 mg/kg) readily suppressed the effects of both cannabinoids and restored the inhibitory response to VTA stimulation. Moreover, when administered alone, SR141716A prolonged the inhibition in 55.6% of the neurons tested. The results indicate that stimulation of CB1 receptors by cannabinoids results in an enhanced excitability of prefrontal cortex pyramidal neurons as indexed by the suppression of the inhibitory effect of VTA stimulation and by the increase in firing rate of antidromically identified neurons projecting to the VTA. Furthermore, our results support the view that endogenous cannabinoids exert a negative control on dopamine activity in the prefrontal cortex. This study may be relevant in helping to understand the influence of cannabinoids on cognitive processes mediated by the prefrontal cortex. PMID- 11488954 TI - Antisense oligodeoxynucleotide-induced suppression of basal forebrain NMDA-NR1 subunits selectively impairs visual attentional performance in rats. AB - It is generally agreed that basal forebrain neuronal circuits contribute to the mediation of the ability to detect, select and discriminate signals, to suppress the processing of irrelevant information, and to allocate processing resources to competing tasks. Rats were trained in a task designed to assess sustained attention, or in a cued discrimination task that did not tax attentional processes. Animals were equipped with guide cannula to infuse bilaterally antisense oligodeoxynucleotides (ODNs) against the N-methyl-D-aspartate (NMDA) NR1 subunits, or missense ODNs, into the substantia innominata of the basal forebrain. Infusions of antisense or missense ODNs did not affect cued visual discrimination performance. Infusions of antisense ODNs dose-dependently impaired sustained attention performance by selectively decreasing the animals' ability to detect signals while their ability to reject nonsignal trials remained unchanged. The detrimental attentional effects of antisense infusions were maximal 24 h after the third and final infusion, and performance returned to baseline 24 h later. Missense infusions did not affect attentional performance. Separate experiments demonstrated extensive suppression of NR1 subunit immunoreactivity in the substantia innominata. Furthermore, infusions of antisense did not produce neurotoxic effects in that region as demonstrated by the Fluoro-Jade METHOD: The present data support the hypothesis that NMDA receptor (NMDAR) stimulation in the basal forebrain, largely via glutamatergic inputs originating in the prefrontal cortex, represents a necessary mechanism to activate the basal forebrain corticopetal system for mediation of attentional performance. PMID- 11488955 TI - Fos imaging reveals differential neuronal activation of areas of rat temporal cortex by novel and familiar sounds. AB - To provide information about the possible regions involved in auditory recognition memory, this study employed an imaging technique that has proved valuable in the study of visual recognition memory. The technique was used to image populations of neurons that are differentially activated by novel and familiar auditory stimuli, thereby paralleling previous studies of visual familiarity discrimination. Differences evoked by novel and familiar sounds in the activation of neurons were measured in different parts of the rat auditory pathway by immunohistochemistry for the protein product (Fos) of the immediate early gene c-fos. Significantly higher counts of stained neuronal nuclei (266 +/- 21/mm2) were evoked by novel than by familiar sounds (192 +/- 17/mm2) in the auditory association cortex (area Te3; AudA). No such significant differences were found for the inferior colliculus, primary auditory cortex, postrhinal cortex, perirhinal cortex (PRH), entorhinal cortex, amygdala or hippocampus. These findings are discussed in relation to the results of lesion studies and what is known of areas involved in familiarity discrimination for visual stimuli. Differential activation is produced by novel and familiar individual stimuli in sensory association cortex for both auditory and visual stimuli, whereas the PRH is differentially activated by visual but not auditory stimuli. It is suggested that this latter difference is related to the nature of the particular auditory and visual stimuli used. PMID- 11488956 TI - D2-like dopamine receptor activation excites rat dorsal raphe 5-HT neurons in vitro. AB - The aim of the present study was to investigate the effect of dopamine (DA) on the excitability of dorsal raphe nucleus (DRN) 5-hydroxytryptamine (5-HT) neurons using the patch-clamp technique in brain slices. Bath application of DA (1-300 microM) produced a concentration-dependent membrane depolarization in all 5-HT neurons examined. This effect persisted in the presence of tetrodotoxin (TTX; 1 microM) and low extracellular calcium. Moreover, blockade of ionotropic glutamate receptors with 6,7-dinitroquinoxaline-2,3-dione (DNQX) and 2-amino-5 phosphonopentanoic acid (AP5) did not prevent DA-induced depolarization, indicating that it was mediated by a direct effect of DA on 5-HT neurons. The DA induced depolarization was not antagonized by selective alpha1-adrenergic receptor antagonists, prazosin and WB 4101, but by a nonselective DA receptor antagonist, haloperidol. In addition, the selective D2-like receptor agonist quinpirole and antagonist sulpiride mimicked and blocked DA-induced depolarization, respectively. These results indicate that DA-induced membrane depolarization in DRN 5-HT neurons is mediated by the activation of D2-like DA receptors. The DA-induced membrane depolarization and inward current were associated with an increase in membrane conductance. Examination of the current voltage (I-V) relationship for the DA-induced inward current revealed that the amplitude of the current increased with membrane hyperpolarization and reversed polarity at a potential near -15 mV. These data suggest that DA-induced depolarization in DRN 5-HT neurons is not mediated by a decrease in potassium conductance, but most likely by the activation of a nonselective cation current. PMID- 11488957 TI - BDNF mRNA expression in rat hippocampus and prefrontal cortex: effects of neonatal ventral hippocampal damage and antipsychotic drugs. AB - Brain-derived neurotrophic factor (BDNF) plays an important role in development, synapse remodelling and responses to stress and injury. Its abnormal expression has been implicated in schizophrenia, a neuropsychiatric disorder in which abnormal neural development of the hippocampus and prefrontal cortex has been postulated. To clarify the effects of antipsychotic drugs used in the therapy of schizophrenia on BDNF mRNA, we studied its expression in rats treated with clozapine and haloperidol and in rats with neonatal lesions of the ventral hippocampus, used as an animal model of schizophrenia. Both antipsychotic drugs reduced BDNF expression in the hippocampus of control rats, but did not significantly lower its expression in the prefrontal cortex. The neonatal hippocampal lesion itself suppressed BDNF mRNA expression in the dentate gyrus and tended to reduce its expression in the prefrontal cortex. These results indicate that, unlike antidepressants, antipsychotics down-regulate BDNF mRNA, and suggest that their therapeutic properties are not mediated by stimulation of this neurotrophin. To the extent that the lesioned rat models some pathophysiological aspects of schizophrenia, our data suggest that a neurodevelopmental insult might suppress expression of the neurotrophin in certain brain regions. PMID- 11488958 TI - Activation of muscarinic receptors induces protein synthesis-dependent long lasting depression in the perirhinal cortex. AB - There is strong evidence that decrements in neuronal activation in perirhinal cortex when a novel stimulus is repeated provide a neural substrate of visual recognition memory. There is also strong evidence that muscarinic acetylcholine (ACh) receptors are involved in learning and memory. However, the mechanisms underlying neuronal decrements in the perirhinal cortex and the basis of ACh involvement in learning and memory are not understood. In an in vitro preparation of rat perirhinal cortex we now demonstrate that activation of ACh receptors by carbachol (CCh) produces long-lasting depression (LLD) of synaptic transmission that is dependent on muscarinic M1 receptor activation. Crucially, the induction of this form of LLD requires neither N-methyl-D-aspartate receptor activation nor synaptic stimulation. CCh-induced LLD was not blocked by the protein kinase C inhibitors staurosporine or BIM, or by the protein phosphatase inhibitor okadaic acid. However, each of cyclopiazonic acid (an agent that depletes intracellular calcium stores) and anisomycin (an inhibitor of protein synthesis) significantly reduced the magnitude of CCh-induced LLD. These mechanisms triggered by muscarinic receptor activation could play a role in the induction and/or expression of certain forms of activity-dependent long-term depression in perirhinal cortex. An understanding of CCh-induced LLD may thus provide clues to the mechanisms underlying lasting neuronal decrements that occur in the perirhinal cortex and hence for neural substrates of visual recognition memory. PMID- 11488960 TI - Chronic psychosocial stress differentially affects apoptosis in hippocampal subregions and cortex of the adult tree shrew. AB - We studied the effect of chronic psychosocial stress on cell death and volume changes in the tree shrew hippocampus. In situ end labelling (ISEL) identified low frequent but convincing apoptosis in many hippocampal subregions. Also in entorhinal cortex, apoptosis was found, generally at higher frequencies. After 28 days of chronic stress, apoptosis was significantly reduced in the CA1 stratum radiatum, whereas an increase was observed in the hilus (P < 0.04). With all subregions taken together, the hippocampus showed a decrease, whereas in the cortex, an increase in apoptosis was found after stress (P < 0.04). In a parallel and similar chronic stress study, post mortem morphometry of the same brain regions was performed, revealing mild decreases (7.6%) in entire hippocampal volume. We conclude that (i) low frequent apoptosis occurs throughout the adult tree shrew brain, and (ii) 28 days of chronic stress differentially affects its occurrence in distinct hippocampal subregions and entorhinal cortex. As previous stereological investigations failed to detect any loss in the principal neuronal layers, psychosocial stress, therefore, must affect other (structural) parameters like dendritic tree, interneurons, neurogenesis, or glia. PMID- 11488959 TI - Roles of the glutamate receptor epsilon2 and delta2 subunits in the potentiation and prepulse inhibition of the acoustic startle reflex. AB - We examined the regulation of the acoustic startle response in mutant mice of the N-methyl-D-aspartate (NMDA)- and delta-subtypes of the glutamate receptor (GluR) channel, which play important roles in neural plasticity in the forebrain and the cerebellum, respectively. Heterozygous mutant mice with reduced GluRepsilon2 subunits of the NMDA receptor showed strongly enhanced startle responses to acoustic stimuli. On the other hand, heterozygous and homozygous mutation of the other NMDA receptor GluRepsilon subunits exerted no, or only small effects on acoustic startle responses. The threshold of the auditory brainstem response of the GluRepsilon2-mutant mice was comparable to that of the wild-type littermates. The primary circuit of the acoustic startle response is a relatively simple oligosynaptic pathway located in the lower brainstem, whilst the expression of GluRepsilon2 is restricted to the forebrain. We thus suggest that the NMDA receptor GluRepsilon2 subunit plays a role in the regulation of the startle reflex. Ablation of the cerebellar Purkinje cell-specific delta2 subunit of the GluR channel exerted little effect on the acoustic startle response but resulted in the enhancement of prepulse inhibition of the reflex. Because inhibition of the acoustic startle response by a weak prepulse is a measure of sensorimotor gating, the process by which an organism filters sensory information, these observations indicate the involvement of the cerebellum in the modulation of sensorimotor gating. PMID- 11488961 TI - Receptive field organization of disparity-sensitive cells in Macaque medial superior temporal cortex. AB - Binocular disparities are crucial for building an accurate three-dimensional representation of the peripersonal environment in a viewer-centred frame of reference. Previous studies have shown that visual cells of the medial superior temporal cortex (MST) have large receptive fields and that they are sensitive to disparities present in large surfaces. By using a reverse cross-correlation technique in this study we tested 175 disparity-sensitive units recorded from MST in the awake Macaca mulatta monkey to determine if these large receptive fields are homogeneous in terms of disparity sensitivity. We found that the receptive fields of 50 cells (50 out of 175, 29%) showed subregions with specific disparity sensitivity. These subregions presented eccentricities from 0.8 to 22.3 degrees and their sizes varied from 1.6 to 15.3 degrees 2. This particular receptive field organization represents a suitable mechanism for encoding the location of small objects within our peripersonal space. PMID- 11488962 TI - Discharge threshold is enhanced for several seconds after a single interictal spike in a model of focal epileptogenesis. AB - Interictal spikes (ISs) are typically observed between seizures in focal epilepsies. Whether ISs are causally involved or represent protective elements in the transition toward an ictal discharge is an open question. Previous studies suggested that inhibition or disfacilitation occurs during the period elapsing between two ISs induced by local application of either bicuculline or penicillin in the piriform cortex of the in vitro isolated guinea pig brain preparation. We further investigated this issue by studying responses to afferent stimulation during the interspike period (6.3 +/- 2.5 s; mean +/- SD). Properly set stimulation intensity of the lateral olfactory tract resets ISs exclusively (and not before) 4-10 s (5.6 +/- 2.0 s; mean +/- SD) after a preceding spontaneous spike. This finding demonstrates the existence of a period of enhanced threshold to stimulus-evoked activation that coincides with the interspike interval in the absence of stimulation. Current source density analysis of depth laminar profiles demonstrated that both stimulus-evoked and spontaneous ISs were generated by the activation of an identical cortical circuit. Our study suggests that interictal spiking could play a protective role or at least provide an effective restraint against the onset of a focal ictal discharge. PMID- 11488963 TI - River basin-related genetic structuring in an endangered fish species, Chondrostoma lusitanicum, based on mtDNA sequencing and RFLP analysis. AB - Chondrostoma lusitanicum is a Portuguese endemic cyprinid with a restricted distribution and reduced numbers in some basins, justifying its status as a threatened species. We examined genetic population structure using samples from throughout its geographical range in Portugal, using sequencing of b cytochrome and restriction fragment length polymorphism analysis of the NADH subunits 5 and 6. There was reduced within-population genetic variability but considerable among population differentiation, particularly marked between both the Mira and Arade basins in the extreme south and other populations. These results confirm phylogeographic relationships suggested by previous fragmentary allozyme studies for C. lusitanicum, and are in accordance with allozyme and mitochondrial DNA data on phylogeography of coexisting cyprinid species of the genus Leuciscus. The levels of genetic divergence revealed by sequence and RFLP data showed strongly concordant patterns: geographical genetic structuring, with the definition of three distinct groups, was observed. The high values of nucleotide divergence and pairwise sequence divergence of the Mira and Arade groups, when compared with all other samples, support a distinct taxonomic status probably at the species level. Results are also discussed in relation to conservation of this highly fragmented species, in terms of Evolutionary Significant Units and Management Units. PMID- 11488964 TI - Genetics of sex determination in the gynodioecious species Lobelia siphilitica: evidence from two populations. AB - In order to determine whether interactions between multiple sex-determining genes might be partly responsible for the wide variation in female frequency among populations of Lobelia siphilitica, we used progeny sex ratios from field collected plants and from controlled crosses within and between two populations. We demonstrate that multiple cytoplasmic male-sterility types are present in a gynodioecious population where female frequency exceeds 50%. These male-sterility types each have corresponding nuclear alleles that can restore pollen fertility. Restoration of one male-sterility type appears to be controlled by a single, dominant allele, but restoration of a second cytoplasmic type is not easily explained with simple genetic models -- perhaps multiple nuclear loci and/or epistatic interactions are involved. In addition, the crosses show that pollen from some hermaphrodite plants in a population containing no females restores male fertility to plants from a geographically distant gynodioecious population that have male-sterile cytoplasm. Furthermore, some plants in that hermaphrodite population carry a male-sterile cytoplasm. Taken together, these results fit theoretical predictions that female frequency might be highly variable among populations when sex is determined by interactions between several nuclear and cytoplasmic genetic factors, some of which may not be present in all populations. The data also illustrate the need for more theoretical and empirical work investigating the evolutionary impact of nuclear restorer genes with complex action, and explaining the existence of nuclear restorers and cytoplasmic male sterility genes in a population where females are very rare. PMID- 11488965 TI - Molecular approaches to the taxonomic position of Peruvian potato cyst nematodes and gene pool similarities in indigenous and imported populations of Globodera. AB - Peruvian potato cyst nematode populations were analysed to assess both their inter- and intraspecific similarities. ITS--RFLP and two satellite DNA sequences were used as taxonomic tools. Both techniques have confirmed that the Peruvian populations have as their closest relatives the European Globodera pallida, despite the detection of clear differences that prevents us from assigning these South American populations unambiguously to any Globodera species. A more precise study of the variability of these Peruvian populations was investigated and they were compared with the imported European populations using protein (2-DGE) and DNA (RAPD) datasets. The clear distinction between the Peruvian and the European populations was confirmed and, inside each group, no correlation was found between the pathotype classification and the observed clustering of the populations. Surprisingly, while RAPDs revealed a higher variability in the Peruvian group than in the European one, some characteristic proteins were found by 2-DGE in some European populations, whereas it was impossible to find any in the Peruvian populations. It is concluded that the primary founders of the European populations may have an origin other than that of the Peruvian populations involved in this study. PMID- 11488966 TI - Population analysis of an unusual NOR-site polymorphism in brown trout (Salmo trutta L.). AB - A population analysis of an unusual NOR-site polymorphism previously detected in brown trout (Salmo trutta L.) from North-western Spain was carried out in 225 individuals from 20 native populations from this area. The analysis performed has permitted us to reveal: (i) the ubiquity of this phenomenon in most river basins from NW Spain, 13 extra-NORs not observed in standard trout being detected; (ii) the rDNA constitution of all extra-NORs, as confirmed by CMA3-staining and rDNA FISH, and their capability to constitute their own nucleolus; (iii) the constant location of extra-NORs within individuals and their stable transmission across generations; and (iv) the telomeric location of Ag-NORs, which were randomly distributed in the karyotype of S. trutta, mostly in the heterozygous condition. Repetitive sequences in the IGS or scattered along rDNA units, rather than chromosome rearrangements, could play an important role in the dispersion of NORs. The polymorphism described seems to have a single geographical origin, since a positive correlation was demonstrated between the degree of polymorphism and geographical distance to a central point of the river basin where the phenomenon showed a higher intensity. The distribution of NOR-site variation as compared with allozyme variation in the populations studied, showed significant statistical differences. Selection against high copy number, or mutation due to changes in NOR location, could be acting on this polymorphism to explain the observed distribution. The high dispersion of NOR-site variants within and among populations suggests the transposition phenomenon responsible for NOR jumping could be still active. PMID- 11488967 TI - Chromosome races with Pliocene origins: evidence from mtDNA. AB - There are eight distinct chromosomal races of the New Zealand weta Hemideina thoracica. We used mtDNA sequence data to test the hypothesis that these races originated on islands during the early Pliocene (7--4 million years ago). Nine major mitochondrial lineages were identified from 65 cytochrome oxidase I sequences. Phylogenetic analysis of these lineages suggests that they arose at approximately the same time. The geographical distribution of some lineages coincides with areas that were islands during the Pliocene. Overall, hierarchical AMOVA analysis shows that chromosomal races and Pliocene islands describe only 28% and 24%, respectively, of the total current mtDNA variation. However, removing one widespread (A) and one putatively introgressed (F) lineage increases these estimates to 65% and 80%, respectively. Intraspecific sequence divergence was very high, reaching a maximum of 9.5% (uncorrected distance) and GC content was high compared to other insect mtDNA sequences. Average corrected distance among mtDNA lineages supports the Pliocene origins of this level of genetic diversity. In the southern part of the species range there is reduced mtDNA variation, probably related to local extinction of H. thoracica populations from recent volcanic activity and subsequent re-colonization from a leading edge. In contrast, in this southern part there are five chromosome races, suggesting that chromosome races here may be younger than those in the north. PMID- 11488968 TI - Genetic and demographic parameters determining population persistence after a discrete change in the environment. AB - Field studies suggest that populations often go extinct following discrete changes in the environment. However, populations may avoid extinction by rapidly adapting to their altered environment. We used a stochastic finite-locus model to estimate the distance the optimal value of a quantitative trait could shift in a single step Delta theta(c) without causing more than 5% of the replicate populations to go extinct. We found that evolution increased the magnitude of Delta theta(c) by at least two phenotypic standard deviations and that such evolution could take place within 5--10 generations. Indeed (Delta theta(c))(2) increased approximately linearly with the logarithm of the initial population size and the rate of this increase was much greater when heritability was high or when stabilizing selection was weak. (Delta theta(c))(2) also increased approximately linearly with the logarithm of per capita fecundity. To our surprise there was no 'demographic rescue' effect from migration; a population augmented with migrants from a neighbouring population where environmental conditions were unchanged was always more likely to go extinct. The addition of mutation, more loci, density-dependence, or environmental stochasticity had only small effects on the outcome. We were able to compare our results for closed populations with density-independent population growth to those from an analytical model and found good agreement so long as the proportion of the offspring surviving selection in the initial generations was at least 1%. PMID- 11488969 TI - Wolbachia endosymbiont responsible for cytoplasmic incompatibility in a terrestrial crustacean: effects in natural and foreign hosts. AB - Wolbachia bacteria are vertically transmitted endosymbionts that disturb the reproduction of many arthropods thereby enhancing their spread in host populations. Wolbachia are often responsible for changes of sex ratios in terrestrial isopods, a result of the feminization of genotypic males. Here we found that the Wolbachia hosted by Cylisticus convexus (wCc) caused unidirectional cytoplasmic incompatibility (CI), an effect commonly found in insects. To understand the diversity of Wolbachia-induced effects in isopods, wCc were experimentally transferred in a novel isopod host, Armadillidium vulgare. wCc conserved the ability to induce CI. However, Wolbachia were not transmitted to the eggs, so the capacity to restore the compatibility in crosses involving two transinfected individuals was lost. The feminizing Wolbachia hosted by A. vulgare was unable to rescue CI induced by wCc. These results showed that Wolbachia in isopods did not evolved broadly to induce feminization, and that CI and the feminizing effect are probably due to different mechanisms. In addition, wCc reduces the mating capacity of infected C. convexus males, suggesting that the bacteria might alter reproductive behaviour. The maintenance of wCc in host populations is discussed. PMID- 11488970 TI - Quantitative trait loci for upper thermal tolerance in outbred strains of rainbow trout (Oncorhynchus mykiss). AB - The expression of three putative QTL for upper thermal tolerance (UTT) was examined in two strains of outbred rainbow trout unselected for this trait using simple-sequence repeat (SSR; microsatellite) markers associated with UTT in backcrosses of lines selected on this trait. Two-way diallel lots in the third generation of an outbred pedigree were exposed to an acute thermal challenge. QTL detection was performed separately by each second-generation parent within each diallel lot, incorporating the effects of full sib families and correlated traits. Inheritance of different alleles at the SSR Ssa20.19NUIG from the sire 93 32-1 was strongly associated with the thermal tolerance of his half sib progeny, explaining 7.5% of their phenotypic variance in this trait. A hierarchical linear model incorporating allelic inheritance from all four grandsires of the experimental diallels (in addition to family specific and covariate trait effects) was also used to detect associations between the SSR and thermal tolerance in their third-generation grandprogeny. Ssa20.19NUIG was strongly associated with thermal tolerance in the grandprogeny of the grandsire G(0)SVM2. The generally stronger marker-trait associations found in male parents may be partially due to reduced chromosomal recombination rates in male salmonids compared to females. These results indicate the effects of a QTL on a fitness related trait in unselected populations of rainbow trout. PMID- 11488971 TI - The zone of sympatry and hybridization of Mytilus edulis and M. galloprovincialis, as described by intron length polymorphism at locus mac-1. AB - Intron-size variation at the actin gene locus mac-1 was used to characterize mussel, Mytilus spp., populations in the approximately 2000-km wide zone of contact and hybridization ('hybrid zone') between M. edulis and M. galloprovincialis in western Europe. Twenty-five samples were collected in 1995 99 in locations within the hybrid zone and from reference populations of each species. We used correspondence analysis on the matrix of allelic frequencies to determine which alleles are characteristic of each species, and to characterize samples along the genetic gradient between M. edulis and M. galloprovincialis. In the hybrid zone, some samples exhibited mac-1 allele frequencies that were typical of M. edulis; other samples were distributed along the M. edulis/M. galloprovincialis gradient and displayed variable levels of intergradation that were not correlated with geography. Some of the latter samples exhibited significant heterozygote deficiencies. The simple admixture hypothesis (Wahlund effect) could not be rejected for two-fifths of the samples. The hybrid zone thus appeared as a mosaic of populations which are either pure M. edulis, or hybrid between M. galloprovincialis and M. edulis, or a mixture of the foregoing with M. galloprovincialis individuals. These results were consistent with published allozyme data, suggesting that they can be extended to the entire nuclear genome. M. edulis mac-1 alleles were present at moderate frequency in Atlantic M. galloprovincialis, and at significantly lower frequency in some Mediterranean samples. This pattern was homogeneous over a broad geographical range within each basin. It was not evident that introgression of M. edulis into M. galloprovincialis presently occurs south of the zone of contact. We propose that the distinctness of the Atlantic M. galloprovincialis population results from past introgression by M. edulis alleles. PMID- 11488972 TI - Sequence variation in the mitochondrial DNA control region of wild African cheetahs (Acinonyx jubatus). AB - Five hundred and twenty-five bp of mitochondrial control region were sequenced and analysed for 20 Acinonyx jubatus and one Felis catus. These sequences were compared with published sequences from another domestic cat, 20 ocelots (Leopardus pardalus) and 11 margays (Leopardus weidii). The intraspecific population divergence in cheetahs was found to be less than in the other cats. However variation was present and distinct groups of cheetahs were discernible. The 80 bp RS2 repetitive sequence motif previously described in other felids was found in four copies in cheetah. The repeat units probably have the ability to form secondary structure and may have some function in the regulation of control region replication. The two central repeat units in cheetah show homogenization that may have arisen by convergent evolution. PMID- 11488973 TI - Genetic variation of morphological traits in Drosophila melanogaster under poor nutrition: isofemale lines and offspring--parent regression. AB - Variation of three morphological traits (thorax length, wing length and sternopleural bristle number) was examined in Drosophila melanogaster reared on a medium with low yeast content and on a standard medium using the isofemale line analysis and offspring--parent regression. The aim was to test whether these experimental approaches give different patterns of changes in genetic variability estimates when stressful and nonstressful environments are compared. Heritabilities and genetic and phenotypic variances were generally higher in the isofemale line design than in the offspring--parent regression design under both standard and poor nutritional conditions. For each trait, the response of heritability to stress was similar in both designs: wing length exhibited lower heritability under poor nutrition, whereas heritabilities of thorax length and sternopleural bristle number did not differ between nutritional regimes. Statistically significant differences in the genetic variances and the environmental variances between stressful and nonstressful environments were recorded only in isofemale lines: the genetic variance of thorax length and the environmental variances of thorax length and wing length were higher under poor nutrition. The results are compared to literature data and possible reasons of increased genetic variability estimates in isofemale lines are briefly discussed. PMID- 11488974 TI - Population structure and dispersal in the Canary Island caddisfly Mesophylax aspersus (Trichoptera, Limnephilidae). AB - Population genetic structure of the circum-Mediterranean caddisfly Mesophylax aspersus (Trichoptera, Limnephilidae) on the Canary Islands was investigated by studying allozyme variation at nine putative loci in five populations. Genetic variability, population structure and gene flow were compared with data in the literature for continental taxa to assess the effect of isolation of island populations on the genetic structure. Larvae were collected from streams on the islands of Tenerife (one population), La Gomera (two populations in the same catchment) and La Palma (two populations in different catchments). Genetic variability within populations was high relative to that recorded previously for continental Trichoptera, e.g. mean heterozygosity was 0.119--0.336 (0.035--0.15 in continental taxa). Highly significant population structuring was observed (mean F(ST)=0.250), and there was significant within-population structuring (mean F(IS)=0.098). Populations from the same catchment or island were no more similar than populations from different islands, which suggests that occasional long distance dispersal, both between and within islands, is the predominant influence on the population structure. This dispersal ability has contributed to the colonization of most permanent streams on the Canary Islands by M. aspersus. PMID- 11488975 TI - Genetic markers substantiate long-term storage and utilization of sperm by female painted turtles. AB - Most studies of genetic parentage in natural populations have been limited to a single breeding season or reproductive episode and, thus, provide only a snapshot of individuals' mating behaviours. Female turtles can store viable sperm in their reproductive tracts for as long as several years, but the extent to which this capacity is utilized in nature has remained unknown. Here, we employ microsatellite markers to assess genetic paternity in successive clutches of individually marked, free-ranging female painted turtles (Chrysemys picta) over a four year period. The genetic data from 113 clutches from this natural population demonstrate that most females (80.5%) remated each year and that each female generally used a single male's sperm to fertilize all clutches laid within a year. However, sperm usage among females varied considerably, and some females apparently used sperm that had been stored for up to three years to fertilize some or all eggs laid in consecutive nesting seasons. Thus, remating by females is not necessary for continued offspring production from a given sire. Furthermore, 13.2% of all clutches examined showed evidence of multiple paternity, and the genetic paternity patterns across years suggest a 'last in, first out' operation of the females' sperm storage tubules. PMID- 11488976 TI - Recruitment of mononuclear leucocytes to osteoarthritic human synovial xenografts in the ears of SCID mice. AB - A system has been established to assess the recruitment of 99mTc hexamethylpropylene amine oxamine (99mTc-HMPAO)-labelled PBMC and [125I]iododeoxyuridine-labelled Con A stimulated lymphoblasts to allogeneic human synovial xenografts in the ears of SCID mice. Successful engraftment of osteoarthritic synovium was achieved in approximately 90% of cases and a connection between the human microvasculature of the xenograft and the circulation of the mouse was shown. Cells were delivered to the xenograft by a system of regional vascular perfusion, thus avoiding the major murine vascular beds. The accumulation of 99mTc-HMPAO-labelled PBMC in mouse ears was monitored in real time. Direct injection of xenograft-bearing ears with recombinant human TNF-alpha, 7 h prior to perfusion, increased the accumulation of both PBMC and lymphoblasts in cytokine-injected ears compared to contralateral control-injected ears. Autoradiography revealed the presence of [125I]iododeoxyuridine-labelled lymphoblasts associated with human microvasculature within the xenograft. However, the increased accumulation of lymphoblasts in cytokine-injected ears occurred in the tissues surrounding the xenograft, where lymphoblasts were associated more often with murine than human vessels. Although the system described offers advantages over similar models, the propensity for mouse endothelium to interact with human leucocytes is likely to be a generic disadvantage for models of human leucocyte recruitment to xenografts in immunodeficient mice. PMID- 11488977 TI - Susceptibility to Leishmania mexicana infection is due to the inability to produce IL-12 rather than lack of IL-12 responsiveness. AB - Almost all inbred mice are highly susceptible to parasites of the Leishmania mexicana complex that includes L. amazonensis and L. mexicana. Recent studies have reported that T cells from L. amazonensis-infected mice fail to respond to IL-12 due to impaired IL-12R expression. Here, we demonstrate that lymph node cells from L. mexicana-infected C57BL/6 and 129Sv/Ev mice respond efficiently to exogenous IL-12 in vitro and produce IFN-gamma. Moreover, we also show that deletion of signal transducer and activator of transcription (STAT)4 gene in resistant STAT6-/- mice renders them susceptible to L. mexicana. These findings indicate that an inability to produce IL-12 rather than unresponsiveness to this cytokine is responsible for susceptibility to L. mexicana. Moreover, the data also demonstrate that the STAT4-mediated pathway is critical for the development of protective immunity against cutaneous leishmaniasis, regardless of the species of Leishmania and/or genetic background of the mice. PMID- 11488978 TI - Lymphotoxin controls alphaEbeta7-integrin expression by peripheral CD8+ T cells. AB - Lymphotoxin (LT)-alpha, a member of the TNF family, is recognized as an important mediator in different aspects of lymphoid organ development. Targeted disruption of this molecule resulted in a substantial reduction in the proportion of alphaEbeta7-integrin(high) CD8+ T cells detectable in peripheral lymphoid organs. This defect, however, was not observed on mature CD4-CD8+ thymocytes. To determine whether this was due to downregulation of beta7-integrin expression by peripheral CD8+ T cells or a failure of thymic emigration of CD8+ beta7 integrin(high) T cells, beta7-integrin was examined on recent thymic emigrants (RTE). When analysed within 16 h after leaving the thymus CD4-CD8+ RTE in both LT alpha-/- and wild type (wt) mice remained beta7-integrin(high) and were indistinguishable. However, within 3-5 days, emigration loss of beta7-integrin became evident in LT-alpha-/- mice. Despite this loss, the proportion of thymically derived alphabetaTCR+ T-cell populations in the intestinal epithelium, an important target tissue of CD8+ alphaEbeta7-integrin(high) T cells, was increased in the absence of LT-alpha. In contrast, B cells were detectable only rarely in the intestinal tissue of LT-alpha-/- mice. The expression of E-Cadherin remained unchanged. These results indicate that a LT-alpha-dependent process maintains a high level of alphaEbeta7-integrin expression by peripheral CD8+ T cells, and with this control mechanism LT-alpha may help to regulate CD8+ T-cell numbers in the tissues. PMID- 11488979 TI - Structural model of human IL-13 defines the spatial interactions with the IL 13Ralpha/IL-4Ralpha receptor. AB - Interleukin-13 (IL-13) plays a key role in immune responses and inflammation. A structural model of human IL-13 (HuIL-13) based on the nuclear magnetic resonance and X-ray structure of IL-4 is put forward. Unlike previous models, this model is based on new sequence alignments that take into account the formation of the two disulfide linkages that have been determined experimentally. The proposed structure of human IL-13 is similar to IL-4, consisting of a four helix bundle with hydrophobic residues lining the core of the molecule and surface polar residues showing a high degree of solvent accessibility. Regions of HuIL-13 that are critical for the interaction with its receptors are explored and discussed in relation to existing mutagenic studies. From these studies we predict that helices A and C of HuIL-13 interact with the IL-4 receptor alpha (IL-4Ralpha) region and helix D is responsible for the interaction with the IL-13 receptor alpha 1 (IL-13Ralpha1) receptor. PMID- 11488980 TI - 'Reverse gear' cellular movement mediated by chemokines. AB - We sought to model the mechanism by which leucocytes may be actively repulsed by a beta-chemokine signal. This model is used to interpret an apparent paradox in chemokine biology, whereby high levels of a T-cell chemoattractant, stromal cell derived factor-1 (SDF-1), are present in bone marrow and thymic tissues despite a paucity of mature T lymphocytes in these areas. We postulate the differential involvement in cell migration of the two binding sites on SDF-1 for its sole receptor, CXCR4, depending on whether high or low concentrations of SDF-1 are encountered by the cell. Site choice would be mediated by divergent affinities of the two binding interactions. We also propose differential signalling following SDF-1/CXCR4 interactions on the plasma membrane versus ligand/receptor complexes in endocytic vesicles. Preliminary data showing divergent susceptibility to kinase inhibitors depending on whether a cell is attracted to or repulsed by SDF 1, are consistent with this model. In terms of physical movement toward or away from a chemokine gradient, we compare the cycling of surface receptors during migration to the caterpillar drive of a tractor, which can change direction simply by altering the direction of rotation of its threads. Finally, the potential clinical implications of concentration-dependent, chemokine-based cell attraction and repulsion are discussed. PMID- 11488981 TI - Cloning and characterization of a mouse homologue of the human haematopoietic cell-specific four-transmembrane gene HTm4. AB - Haematopoietic cell-specific transmembrane-4 (HTm4) is a four-transmembrane protein most closely related to CD20 and the beta subunit of the high affinity receptor for IgE (Fc(epsilon)RIbeta). To date, it has only been described in humans, where it is expressed in haematopoietic cells of both myeloid and lymphoid lineages. The function of HTm4 is unknown; however, as for CD20 and Fc(epsilon)RI-beta, it is likely to play a role in signal transduction as part of a multi-subunit cell surface receptor complex. In this study, we report the cDNA cloning and expression distribution of mouse HTm4. The deduced mouse HTm4 protein is of 213 amino acids, and contains four putative transmembrane domains. Mouse HTm4 shows 62% overall amino acid identity with human HTm4; the transmembrane regions are highly conserved between both species (75% identity), whereas the N- and C-terminal and inter-transmembrane loop regions are more divergent (52%). Interestingly, the N-terminal domain of mouse HTm4 is predicted to be 23 amino acids shorter, and the C-terminal domain 23 amino acids longer, than that of human HTm4. Northern blot and reverse transcriptase (RT)-PCR analysis suggest that mouse HTm4 mRNA is expressed at low levels only in spleen, bone marrow and peripheral blood leucocytes. This is the first report of the cloning of HTm4 from a species other than human, and provides important sequence information towards the understanding of the function of this poorly characterized four-transmembrane molecule. PMID- 11488982 TI - Towards a unified model of neuroendocrine-immune interaction. AB - Although the neuroendocrine system has immunomodulating potential, studies examining the relationship between stress, immunity and infection have, until recently, largely been the preserve of behavioural psychologists. Over the last decade, however, immunologists have begun to increasingly appreciate that neuroendocrine-immune interactions hold the key to understanding the complex behaviour of the immune system in vivo. The nervous, endocrine and immune systems communicate bidirectionally via shared messenger molecules variously called neurotransmitters, cytokines or hormones. Their classification as neurotransmitters, cytokines or hormones is more serendipity than a true reflection of their sphere of influence. Rather than these systems being discrete entities we would propose that they constitute, in reality, a single higher-order entity. This paper reviews current knowledge of neuroendocrine-immune interaction and uses the example of T-cell subset differentiation to show the previously under-appreciated importance of neuroendocrine influences in the regulation of immune function and, in particular, Th1/Th2 balance and diurnal variation there of. PMID- 11488983 TI - In vitro induction of CD25+ CD4+ regulatory T cells by the neuropeptide alpha melanocyte stimulating hormone (alpha-MSH). AB - Recently, we have found that the neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH) not only suppresses IFN-gamma production, but also induces TGF-beta1 production by activated effector T cells. These alpha-MSH- treated effector T cells function as regulatory T cells in that they suppress IFN-gamma production and hypersensitivity mediated by other effector T cells. Experimental autoimmune uveoretinitis (EAU) was suppressed in its severity and incidence in mice that were injected with primed T cells activated in vitro by APC and antigen in the presence of alpha-MSH. Moreover, it appeared that alpha-MSH had converted a population of effector T cells polarized to mediate hypersensitivity into a population of T cells that now mediated immunoregulation. To characterize these alpha-MSH- treated T cells, primed T cells were TCR-stimulated in the presence of alpha-MSH in vitro and their lymphokine profile was examined. Such effector T cells displayed enhanced levels of TGF-beta1 production and no IFN-gamma or IL 10, with IL-4 levels remaining unchanged in comparison with inactivated T cells. In addition, if soluble TGF-beta receptor II was added to cocultures of alpha-MSH treated T cells and activated Th1 cells, the alpha-MSH-treated T cells could not suppress IFN-gamma production by the Th1 cells. These results suggest that alpha MSH induces T cells with a regulatory lymphokine pattern, and that through their production of TGF-beta1 these cells suppress other effector T cells. Examination of the alpha-MSH-treated T cells showed that alpha-MSH did not alter the phosphorylation of CD3 molecules following TCR engagement. Primed T cells express the melanocortin 5 receptor (MC5r), a receptor that is linked to an intracellular signalling pathway shared by other cytokine receptors. Blocking the receptor with antibody prevented alpha-MSH from suppressing IFN-gamma production by the activated regulatory T cells, suggesting that alpha-MSH immunoregulation is through the MC5r on primed T cells. Surface staining and cell sorting of the alpha-MSH- treated primed T cells showed that the regulatory T cells are CD25+ CD4+ T cells. From these results we find that alpha-MSH can mediate the induction of CD25+ CD4+ regulatory T cells. These regulatory T cells require specific antigen for activation, but through non-specific TGF-beta1-mediated mechanisms they can suppress other effector T cells. PMID- 11488984 TI - Neuroendocrine properties of macrophage migration inhibitory factor (MIF). AB - The cytokine macrophage migration inhibitory factor (MIF) is produced by neuroendocrine and immune tissues and possesses several features that allow it to be characterized as a neuroendocrine mediator. Its pro-inflammatory action and its pathogenic role in inflammatory diseases, such as septic shock, arthritis and other diseases, have clearly been demonstrated and may be based in part on neuroendocrine mechanisms. Macrophage migration inhibitory factor possesses glucocorticoid-antagonist properties within the immune system and participates in the regulation of several endocrine circuits. This review summarizes the current state of MIF research and focuses on MIF expression and function in nervous and endocrine tissues. PMID- 11488985 TI - Cross-talk between pro-inflammatory transcription factors and glucocorticoids. AB - Asthma is a chronic inflammatory disease of the airway that is characterized by cellular infiltration and activation. These processes are induced by overexpression of chemokines and cytokines, such as eotaxin, IL-1beta and GM-CSF. These mediators are downstream targets for the transcription factors activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB), which control the expression of most immunomodulatory genes and whose activity and expression are elevated in asthma. Glucocorticoids are the most effective anti-inflammatory drugs used in the treatment of chronic inflammatory diseases such as asthma. They act by binding to a specific glucocorticoid receptor (GR) that on activation translocates to the nucleus and either increases (transactivates) or decreases (transrepresses) the expression of responsive genes. Transrepression is the major mechanism of glucocorticoid action in inhibiting inflammatory gene expression. Thus, the ability of the transcription factors AP-1 and NF-kappaB to induce gene transcription is attenuated by GR. Although only 5-10% of asthmatic subjects are glucocorticoid-insensitive, these subjects account for over 50% of the health care costs for asthma (> $6 billion per annum). Examining these patients also gives an insight into important aspects of glucocorticoid action in controlling inflammation and into the development of potential new drugs. Biochemical and genomic studies have indicated abnormal induction of the c-Jun N-terminal kinase (JNK) pathway in some of these patients. The ability of most patients to respond to dexamethasone with induction of histone acetylation correlated with nuclear translocation of GR. However, a subgroup of these patients had an inability to correctly interact with the basal transcription complex in spite of high levels of nuclear GR. This suggests that cross-talk between pro- and anti-inflammatory transcription factors may modulate activation of the transcriptional complex and thereby reduce steroid actions. PMID- 11488986 TI - Transcription factor-mediated molecular mechanisms involved in the functional cross-talk between cytokines and glucocorticoids. AB - After antigenic stimulation the increase in cytokine levels constitutes a fundamental event in the host defense and mediates many processes such as inflammation, B- and T-cell growth and differentiation and activation of effector cells. Most of these processes depend on the cytokine-induced activation of transcription factors that modulate the expression of target genes. Cytokines induce a rise in glucocorticoid levels, which are instrumental in controlling immune-cytokine overreactions. Because of their anti-inflammatory and immunosuppressive actions, glucocorticoids are highly useful as therapeutic drugs in a range of diseases. The cross-talk between cytokine-induced transcription factors such as nuclear factor-kappaB, activating protein-1, cAMP responsive element binding protein and nuclear factor of activated T cells, and glucocorticoid receptors involves both genomic and non-genomic actions, and constitutes the mechanism by which glucocorticoid repressive effects on cytokine synthesis and action take place. These molecular interactions represent the key for the study of physiological compensatory actions of corticosteroids, the interactions of cytokines and glucocorticoids at their target cells, as well as the therapeutic benefits and side-effects of synthetic steroids. For this reason, we will focus on the molecular aspects of cytokine-glucocorticoid interactions, represented by the cross-coupling between cytokine-mediated transcription factors and glucocorticoid receptors. PMID- 11488987 TI - Hypothalamic-pituitary-adrenal axis regulation of inflammation in rheumatoid arthritis. AB - The profound anti-inflammatory effects of glucocorticoids in drug therapy are reflected in the effects in vivo of endogenous glucocorticoids produced by the adrenals. The production of adrenal glucocorticoids is driven by the hypothalamus and pituitary, which in turn are responsive to circulating products of the inflammatory response, especially cytokines. That inflammation can drive the production of anti-inflammatory glucocorticoids denotes the hypothalamic pituitary-adrenal (HPA)-immune axis as a classic negative feedback control loop. Defects in HPA axis function are implicated in susceptibility to, and severity of, animal models of rheumatoid arthritis (RA), and are hypothesized to contribute to the human disease. In this paper, data supporting the concept of the HPA axis as a regulator of the inflammatory response in animal models of arthritis are reviewed, along with data from studies in humans. Taken together, these data support the hypothesis that the HPA axis provides one of the key mechanisms for inhibitory regulation of the inflammatory response. Manipulation of HPA axis-driven endogenous anti-inflammatory responses may provide new methods for the therapeutic control of inflammatory diseases. PMID- 11488988 TI - Neuroendocrine-immune (NEI) circuitry from neuron-glial interactions to function: Focus on gender and HPA-HPG interactions on early programming of the NEI system. AB - Bidirectional communication between the neuroendocrine and immune systems during ontogeny plays a pivotal role in programming the development of neuroendocrine and immune responses in adult life. Signals generated by the hypothalamic pituitary-gonadal axis (i.e. luteinizing hormone-releasing hormone, LHRH, and sex steroids), and by the hypothalamic-pituitary-adrenocortical axis (glucocorticoids (GC)), are major players coordinating the development of immune system function. Conversely, products generated by immune system activation exert a powerful and long-lasting regulation on neuroendocrine axes activity. The neuroendocrine immune system is very sensitive to preperinatal experiences, including hormonal manipulations and immune challenges, which may influence the future predisposition to several disease entities. We review our work on the ongoing mutual regulation of neuroendocrine and immune cell activities, both at a cellular and molecular level. In the central nervous system, one chief compartment is represented by the astroglial cell and its mediators. Hence, neuron-glial signalling cascades dictate major changes in response to hormonal manipulations and pro-inflammatory triggers. The interplay between LHRH, sex steroids, GC and pro-inflammatory mediators in some physiological and pathological states, together with the potential clinical implications of these findings, are summarized. The overall study highlights the plasticity of this intersystem cross-talk for pharmacological targeting with drugs acting at the neuroendocrine-immune interface. PMID- 11488989 TI - Molecular biology and pathogenesis of the human T-cell leukaemia/lymphotropic virus Type-1 (HTLV-1). AB - Retroviruses are associated with a variety of diseases, including immunological and neurological disorders, and various forms of cancer. In humans, the Human T cell Leukaemia/Lymphotropic virus type 1 (HTLV-1), which belongs to the Oncovirus family, is the aetiological agent of two diverse diseases: Adult T-cell leukaemia/lymphoma (ATLL) (Poiesz et al. 1980; Hinuma et al. 1981; Yoshida et al. 1982), as well as the neurological disorder tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM) (Gessain et al. 1985; Rodgers-Johnson et al. 1985; Osame et al. 1986). HTLV-1 is the only human retrovirus known to be the aetiological agent of cancer. A genetically related virus, HTLV-2, has been identified and isolated (Kalyanaraman et al. 1982). However, there has been no demonstration of a definitive aetiological role for HTLV-2 in human disease to date. Simian T-cell lymphotropic viruses types 1 and 2 (STLV-1 and -2) and bovine leukaemia virus (BLV) have also been classified in same group, Oncoviridae, based upon their similarities in genetic sequence and structure to HTLV-1 and -2 (Burny et al. 1988; Dekaban et al. 1995; Slattery et al. 1999). This article will focus on HTLV-1, reviewing its discovery, molecular biology, and its role in disease pathogenesis. PMID- 11488992 TI - Effects of hyaluronan on the invasive properties of human breast cancer cells in vitro. AB - Hyaluronan (HA) is a high molecular weight glycosaminoglycan present mostly in the extracellular matrix (ECM). HA binds to specific receptors such as CD44. Its production is increased at the tumour-stroma interface, including those in breast cancer tumours. It has been suggested that it facilitates invasion of tumour cells into the ECM by a hydrodynamic effect, or by altering tumour cell behaviour. Using in vitro tests we studied the effect of immobilized (iHA) and soluble (sHA) HA on the invasive properties of four human breast cancer cell lines with different levels of CD44 expression. Our results show that iHA acts as an adhesive, haptotactic, and motility stimulating factor for the CD44 positive Hs578T cells and induces the expression of membrane CD44. sHA also changes the motility properties of the Hs578T and MDA-231 cells and increases their CD44 expression. sHA or iHA have no measurable effect on the adhesion, motility or CD44 expression of the ZR-75-1 and MCF-7 breast cancer cells. Our results establish that in high CD44 expressing breast cancer cells HA modulates tumour cell adhesion and motility and also increases the expression of its own receptor, CD44. PMID- 11488993 TI - Production and distribution of interleukin 15 and its receptors (IL-15Ralpha and IL-R2beta) in the implant interface tissues obtained during revision of failed total joint replacement. AB - Failure of total joint replacement (TJR) is a major problem and it is estimated that 15-20% of TJR will fail within 5-10 years after implantation. Most TJR is attributed to aseptic loosening of the implants in association with resorption of related bone due to the release of bone-associated cytokines. IL-15 is a cytokine that activates T cells and natural killer (NK) cells. IL-15 protein is ubiquitous and is expressed in many tissues and cell types. Using immunohistochemical techniques, we demonstrated the expression of IL-15 and its receptors IL-15Ralpha and IL-2Rbeta in the interface tissues obtained from revision surgery. Both IL-15 protein and IL-15Ralpha were observed in macrophages, multinucleated giant cells and endothelial cells around blood vessels. Both the SDS-PAGE and western blot revealed multiple bands and after stages of glycosylation, this resulted in a band at 13 KDa which corresponds to the IL-15 protein. Again RT-PCR results demonstrated a band of 420 bp corresponding to the IL-15 protein. In addition, using U937 cells, the expression of both IL-15 protein and IL-15Ralpha were considerably up-regulated when challenged with retrieved metal particles. Our results illustrated the IL-15 to be an intact protein and that it is stored in the cytoplasm. A dye exclusion cell viability test displayed an increase in toxicity with an increase in the amount of metal particles added. There was a discrepancy between abundant IL-15 mRNA, intracellularly detectable IL-15 protein and apparently inefficient secretion. This suggests that IL-15 protein production is predominantly regulated post-transcriptionally and this is indicated by its strict regulation, especially at cell trafficking. Finally, unlike IL-2, IL-15 plays a certain role in bone resorption that leads to failed joint prostheses. It is apparent that this cytokine is an important T cell mediated immune response which needs further research. PMID- 11488995 TI - Ovarian cancer screening in the general population: current status. AB - There has been considerable interest in the prospect of early detection of ovarian cancer through screening asymptomatic women from the general population. The results of recently published trials provide encouraging data on the possible impact on mortality. There are, however, concerns about compliance, cost, and morbidity from surgery for false positive screen results. The two ongoing randomized controlled trials in the US and UK aim to establish definitively the true impact of screening on ovarian cancer mortality while comprehensively tackling the issues of target population, compliance, health economics, and physical and psychological morbidity of screening. PMID- 11488990 TI - Epstein-Barr virus infection and human malignancies. AB - The Epstein-Barr virus (EBV) is a herpes virus which establishes a life-long persistent infection in over 90% of the human adult population world-wide. Based on its association with a variety of lymphoid and epithelial malignancies, EBV has been classified as a group 1 carcinogen by the International Agency for Research on Cancer. In this article we discuss the evidence supporting an aetiological role for EBV in the pathogenesis of human tumours. The biology of EBV infection will be described with special emphasis on viral transforming gene products. A brief survey of EBV-associated tumours is followed by a discussion of specific problems. Evidence is presented which suggests that failures of the EBV specific immunity may play a role in the pathogenesis of EBV-associated tumours also in patients without clinically manifest immunodeficiencies. Finally, the timing of EBV infection in the pathogenesis of virus-associated malignancies is discussed. There is good evidence that EBV infection precedes expansion of the malignant cell populations in some virus-associated tumours. However, this is clearly not always the case and for some of these tumours there are indications that clonal genetic alterations may occur prior to EBV infection. Thus, whilst there is good evidence to suggest that EBV is a human carcinogen, its precise role(s) in the development of virus-associated human tumours requires clarification. PMID- 11488991 TI - The functions of cytokines and their uses in toxicology. AB - Cytokines are critical controllers of cell, and hence tissue, growth, migration, development and differentiation. The family includes the inflammatory cytokines such as the interleukins and interferons, growth factors such as epidermal and hepatocyte growth factor and chemokines such as the macrophage inflammatory proteins, MIP-1alpha and MIP-1beta. They do not include the peptide and steroid hormones of the endocrine system. Cytokines have important roles in chemically induced tissue damage repair, in cancer development and progression, in the control of cell replication and apoptosis, and in the modulation of immune reactions such as sensitization. They have the potential for being sensitive markers of chemically induced perturbations in function but from a toxicological point of view, the detection of cytokine changes in the whole animal is limited by the fact that they are locally released, with plasma measures being generally unreliable or irrelevant, and they have short half lives which require precise timing to detect. Even where methodology is adequate the interpretation of the downstream effects of high, local concentrations of a particular cytokine is problematic because of their interdependence and the pleiotropism of their action. A range of techniques exist for their measurement including those dependent upon antibodies specific for the respective cytokines, but with the introduction of genomic and proteomic technology, a more complete study of cytokine changes occurring under the influence of chemical toxicity should be possible. Their further study, as markers of chemical toxicity, will undoubtedly lead to a greater understanding of how synthetic molecules perturb normal cell biology and if, and how, this can be avoided by more intuitive molecular design in the future. PMID- 11488996 TI - The role of molecular biology in understanding ovarian cancer initiation and progression. AB - In this review, we will present clinical and experimental data that the surface epithelial cells of the ovary are the most likely cell of origin of ovarian cancer. Using a rat model of the disease, we demonstrate the utility of the molecular techniques of Differential Display Genome Scanning and Suppression Subtractive Hybridization to detect gene expression and genetic differences between normal rat surface epithelial cells and their transformed counterpart. Lastly, we provide examples of how molecular techniques can be used to predict which tumors will respond to chemotherapy. PMID- 11488997 TI - Early stage ovarian cancer: the Italian contribution to clinical research. An update. AB - Early ovarian cancer (stages IA-IIA) accounts for 30% of all epithelial ovarian cancer. Even if relatively uncommon, when "high risk" patients are considered, it is lethal in 25-30% of the cases. Mainstay of treatment is surgery followed by either adjuvant chemotherapy or radiotherapy when indicated on the basis of still debatable prognostic factors. Literature data show a great variability in survival rate due to the great heterogeneity of patients considered in different reports and few randomized trials affected by a consequent low power. Italian groups have contributed both in investigating the role of surgery and of chemo or radiotherapy in the treatment of this disease. An important contribution in surgery has been made by Italian institutions in reducing the extent of surgery in young patients wishing to retain their reproductive capability showing that a "conservative surgery" (unilateral oophorectomy) can be safely performed in initial stages without affecting the probability of cure. Another important surgical topic investigated by Italian institutions concerns the role of lymphadenectomy. In early ovarian cancer the node involvement ranges between 14 24% in stage I and 37-50% in stage II. Although the node positivity rate detectable by sampling (SA) is lower than the one shown by a systematic procedure (LY), no data at the moment show that patients undergoing a sampling evaluation have a poorer prognosis. From 1992 through 1994, 202 patients (SA: 99; LY: 103) were enrolled by six Italian institutions in a randomized trial aimed to assess the diagnostic and therapeutic role of SA vs. LY in early stage ovarian cancer. Positive nodes were detected in 9.9% vs. 19.3% respectively as well as a different proportion of intra/perioperative complications occurred. No difference in time to relapse nor in overall survival were detected in the two groups showing no evidence of efficacy in favor of extensive staging of the retroperitoneum. From 1983 to 1990, 271 stage I ovarian cancer patients entered two prospective multicentric randomized trials conducted by Italian institutions. Trial I compared cisplatin (50 mg/m2, six cycles repeated every 28 days) vs. no further treatment in stage IA-B grade 2-3 patients; Trial II compared the same dose and schedule of cisplatin vs. intraperitoneal P32 in stage IC patients. Cisplatin significantly reduced the relapse rate by 65% in Trial I and by 61% in Trial II, but survival was not affected (Trial I: HR = 1.15, 95% CI = 0.44-2.98; Trial II: HR = 0.72, 95% CI = 0.37-1.43). The final conclusion drawn by these two important Italian studies was that adjuvant cisplatin treatment in early ovarian cancer prevents relapse although the impact of chemotherapy remains unclear. For this reason two international trials have been performed (ICON1 and ACTION) aimed at assessing the role of platinum-based chemotherapy on survival. Italian collaboration in both trials has been important, including about half of the total number of the 900 randomized patients. Results will probably be available during this year and are expected with a great interest by the whole scientific international community. PMID- 11488999 TI - The integration of paclitaxel and new platinum compounds in the treatment of advanced ovarian cancer. AB - There has been a steady improvement in the survival of patients with advanced ovarian cancer. This has been the result of a more skilled surgical approach to these patients and the development of more effective chemotherapy with a better integration of both modalities in first-line treatment. The current optimal chemotherapeutic approach consists of a platinum compound together with paclitaxel. This recommendation is based upon level I evidence of two large randomized trials which established that the combination of paclitaxel-cisplatin was superior to cyclophosphamide-cisplatin. The long-term follow-up of one of these studies continues to show a significant difference in survival at 5 years. Neurotoxicity has been problematic with these regimens, in particular when paclitaxel was given with the higher dosed shorter infusion schedule, as was done in the European-Canadian Intergroup study. Several approaches to reduce this toxicity have been studied. Among these are the use of different paclitaxel infusion schedules, and the application of less neurotoxic platinum compounds. Weekly paclitaxel has a different toxicity profile than the higher dosed three weekly schedules, with less neutropenia, alopecia, arthralgia and neurotoxicity. Four platinum compounds are currently marketed: cisplatin, carboplatin, oxaliplatin, and nedaplatin. Of these only cisplatin, carboplatin, and nedaplatin have been approved for the treatment of patients with ovarian cancer (nedaplatin only in Japan). The equivalence of carboplatin and cisplatin has been suggested from trials without a taxoid. Three randomized studies of paclitaxel-carboplatin vs. paclitaxel-cisplatin concluded that paclitaxel-carboplatin is the preferred regimen in terms of (less) toxicity and, where studied, in terms of quality of life. So far, no difference in response rates or progression-free survival has been shown. More mature data on overall survival are awaited. Oxaliplatin (a diaminocyclohexane platinum compound) is of interest because it is only partially cross-resistant with cis- or carboplatin and devoid of severe bone marrow suppression, nephrotoxicity, or ototoxicity. Its dose-limiting toxicity is an unusual form of sensory neuropathy, which is cumulative and, contrary to cisplatin's neurotoxicity, generally reversible. Combinations with other active standard agents, as well as platinum compounds and/or taxoids, are feasible and have shown interesting activity. Similar to carboplatin and oxaliplatin, nedaplatin (cis-diammineglycolatoplatinum) can be given without hydration; its dose-limiting toxicity is myelosuppression, in particular thrombocytopenia. Although activity has been shown, no data from randomized comparative trials are available to allow a judgement on its potential advantages. PMID- 11489000 TI - The integration of docetaxel into first-line chemotherapy for ovarian cancer. AB - Docetaxel is being explored as an alternative to paclitaxel in the treatment of ovarian cancer for several reasons: a) evidence of superiority in preclinical models; b) at least comparable activity in platinum-refractory patients (28% response rate in four pooled Phase II trials), together with activity (23% response rate) in paclitaxel-refractory patients; c) indirect evidence of superiority in breast cancer; d) easier administration, i.e., 1 h q3 week schedule vs. 3 or 24 h infusions; and e) potentially superior toxicity profile, particularly regarding neurotoxicity. The Scottish Gynaecological Cancer Trials Group (SGCTG) has performed successive first-line feasibility trials of docetaxel in combination with cisplatin (100 patients) and carboplatin (141 patients). For docetaxel/carboplatin, a regimen of 75 mg/m2 and AUC 5 proved optimal. Over 90% of patients completed six cycles, q3 weekly, and toxicity was very acceptable; a low level of neurotoxicity (5%) was particularly noteworthy, since levels of over 30% are regularly reported for paclitaxel-carboplatin. Activity comparable to paclitaxel-carboplatin (median progression free survival of 16 months) therefore justified a randomized comparison between the two regimens (with paclitaxel 175 mg/m2 in 3 h and carboplatin AUC 5). This has now been completed, with 1077 patients (FIGO stage IC-IV disease) randomized, from 83 centers in 10 countries. Accrual was accomplished in 17 months (October 1998 to May 2000). A toxicity analysis has been completed, since the last patient finished treatment in October 2000. Treatment was delivered as prescribed (6 cycles) in a similar number of patients (79-84%). Significant differences in toxicity were seen, and this analysis together with response data is scheduled for presentation at the May 2001 ASCO meeting. PMID- 11489001 TI - The integration of anthracyclines in the treatment of advanced ovarian cancer. AB - Since the publication of the Gynecologic Oncology Group (GOG) protocol 111 in 1996, and the results of the Arbeitgemeinschaft Gyna kologische Onkologie (AGO) trial Ovar-3 and the GOG protocol 158, the combination of platinum and paclitaxel has been adopted as the standard therapy in advanced ovarian cancer. One option for achieving further progress in the first-line treatment of advanced ovarian cancer might be the addition of noncross-resistant drugs to the two-drug regimen. Meta-analysis showed a survival benefit for platinum-anthracycline based combinations as compared to platinum-based combinations without anthracyclines. An AGO phase I/II trial compared epirubicin in combination with carboplatin and paclitaxel in untreated patients with gynecological malignancies. Based on the results of this study a randomized phase III trial together with the French GINECO group was conducted. The trial started 11/97 and was closed 11/99. All 1281 patients were randomized. Currently, 1132 end-of-therapy reports have been issued. Nine hundred eighty nine (87%) patients completed six cycles of treatment. Treatment and toxicity data are available for these patients. Three hundred thirty five patients had a measurable residual tumor after initial debulking surgery. Response data of 228 patients (111 ET-Carbo, 117 Carbo-T) are available. PMID- 11489002 TI - Gemcitabine in the treatment of ovarian cancer. AB - Gemcitabine is a nucleoside antimetabolite with established activity against several solid tumors. The activity of the drug in patients with ovarian cancer has been reviewed both in patients who have received single drug treatment and in patients who have received combination chemotherapy. The response rates, with single agent gemcitabine, range from 13 to 24% both in previously treated and untreated patients. Doublets consisting of gemcitabine-cisplatin or gemcitabine paclitaxel, in previously treated patients, induced response in 53% and 40% of the patients, respectively. In three studies, first-line treatment with the combination of cisplatin and gemcitabine induced remission in 53% to 71% of the patients. The triplet, including gemcitabine, paclitaxel, and cisplatin or carboplatin, has been examined in previously treated patients and a response rate of 100% was observed. In previously untreated patients the combination of gemcitabine, paclitaxel, and carboplatin has been preferred due to a more favorable toxicity profile. The activity of this combination, observed in 25 evaluable patients, was very high as all patients responded. Complete remission was observed in 60% of the patients and partial remission in 40%. Based on these promising data the triplet consisting of gemcitabine, paclitaxel, and carboplatin has been included in randomized trials both in the US and in Europe. PMID- 11489003 TI - Developmental chemotherapy in advanced ovarian cancer: incorporation of topoisomerase-I inhibitors and perspective of the Gynecologic Oncology Group. AB - Despite improvements in median and overall survival using a combination of platinum and paclitaxel, long-term survival rates for patients with advanced epithelial ovarian carcinoma (EOC) remain disappointing, and the development of more effective primary therapy remains a priority. In particular, several interesting chemotherapy agents have demonstrated activity individually in patients with recurrent EOC. Among these are gemcitabine, topotecan, liposomal doxorubicin, and prolonged oral etoposide. Preclinical models have suggested an advantage for combinations of these agents with platinum, which has been attributed to inhibition of DNA synthetic pathways involved in the repair of platinum-DNA adducts. However, efforts to develop multidrug combinations with platinum and paclitaxel have encountered substantial bone marrow toxicity, prompting exploration of alternative schedules and sequences of drug administration. In this regard, the Gynecologic Oncology Group (GOG) has conducted a series of phase I pilot studies in previously untreated patients to define combinations that are suitable for group-wide phase III trials. With international collaboration, GOG has launched a five-arm trial (GOG-0182) that will compare these combinations against carboplatin-paclitaxel. The selection of candidate regimens for this trial illustrate the challenges of drug development in EOC. PMID- 11489004 TI - Second-line treatment and consolidation therapies in advanced ovarian cancer. PMID- 11489005 TI - Intraperitoneal hyperthermic perfusion chemotherapy of patients with chemotherapy resistant peritoneal disseminated ovarian cancer. AB - The purpose of this article is to evaluate in a prospective, open-label clinical trial the feasibility and efficacy of intraperitoneal hyperthermic perfusion chemotherapy (IPHC) on the survival and quality of life of patients with advanced, peritoneal disseminated ovarian cancer. Thirty-six patients with ovarian cancer were accrued for the study, their selection being based on their progression following different systemic therapies with anti-neoplastic (multiple chemotherapy-resistant or -refractory) agents. The average number of chemotherapy cycles given before the first IPHC was 12.5. The patients' average Karnofsky performance status was 60% and 17 out of 36 patients had ascites before IPHC. The input temperature of the solution for abdominal lavage was 48-49 degrees C: the intraperitoneal temperature was 42-43 degrees C. The flow-rate of the solution for heat exchange was 190-220 ml/min with treatment lasting 1 h at temperatures greater than or equal to 42 degrees C. Median overall survival time (MOS) from first diagnosis of disease (1stDx) was 49 +/- 8 months and from the first IPHC treatment 19 +/- 4 months. The observed 1-year overall survival rate (OSR) of all patients from the start of the first IPHC was 65 +/- 8% and the 5-year OSR was 16 +/- 7%. Malignant ascites vanished within less than 3-5 IPHCs. Quality of life could be improved. The adverse effects were mild especially compared to systemic chemotherapy. In 3 out of 162 treatments, peritoneal disturbances with symptoms of subileus were observed. We conclude that IPHC is technically feasible, safe, and associated with a marked prolongation of survival and improvement in quality of life. Even heavily pretreated patients could be treated safely. Some patients did respond to IPHC even after 25 IPHC treatments. From these results, it can be concluded that IPHC may also improve the treatment outcome of patients with ovarian cancer as salvage therapy, in second-line treatment or even as consolidation or maintenance therapy following induction chemotherapy to patients with suboptimal stage III and IV disease. This should be demonstrated in randomized controlled studies. PMID- 11489006 TI - High dose chemotherapy in ovarian cancer. PMID- 11489007 TI - Impact of new non-cytotoxics in the treatment of ovarian cancer. AB - Over the last decade, a number of new cytotoxic chemotherapy agents have shown evidence of antitumor activity in patients with ovarian carcinoma. These agents are currently being evaluated in large multinational randomized trials to determine whether their addition either concurrently or sequentially to standard paclitaxel and carboplatin regimens will result in improved survival. Whether these new combinations will provide additional benefit may be uncertain; however, it is certain that additional toxicity will limit the continued evaluation of the strategy of adding cytotoxics together. New approaches to improve the systemic therapy of ovarian cancer need to be explored. The next decade will see many trials of non-cytotoxics having a wide range of subcellular and extracellular targets. Many of these targets are abnormally expressed in a variety of solid tumors; thus, it is expected that many of these agents will be appropriate to evaluate in patients with ovarian carcinoma. Based on promising data from preclinical and early clinical studies as well as the presumed applicability of these targets to ovarian carcinoma, the inhibitors of growth factor receptors such as epidermal growth factor receptor and inhibitors of angiogenesis are of particular interest. Despite the interest of the investigators, the rapid evaluation of these target-specific non-cytotoxics is limited by the lack of accurate information on the expression of target in ovarian tumors and the relevance of target expression and its modulation to this tumor type. Early clinical trials are being designed to address these concerns; however, the clinical impact of non-cytotoxic agents in epithelial ovarian carcinoma patients must await the completion of randomized evaluations in combination with standard chemotherapeutic regimens. PMID- 11489008 TI - Intergroup collaboration in ovarian cancer: the Gynecologic Cancer Intergroup (GCIG). AB - Randomized clinical trials are considered the definitive source of evidence for guiding decisions in clinical practise. In patients with ovarian cancer many of the randomized trials from the 1970s and 1980s did not have the statistical power to detect realistic and clinically important differences. Therefore, evolution of so-called "optimal therapy" has been rather slow and some important questions have gone unanswered. In order to overcome this challenge, representatives of different Cooperative Groups started an Ovarian Cancer Trials Intergroup Network in the mid 1990s, as a vehicle to develop future trial collaboration and as a mechanism for communication about strategic directions of phase I/II trials within individual Groups. In 1997 this network became more formalized, going beyond ovarian cancer as a single focus, and the Gynecologic Cancer Intergroup (GCIG) was created. At present GCIG includes representatives of 12 Cooperative Groups and the NCI-US, and functions as a forum for development of global trial collaborations, which hopefully will answer important questions in a more timely fashion and make them more rapidly available for the oncologic community. PMID- 11489009 TI - Remarks and conclusions on ovarian cancer treatment. AB - Ovarian cancer is still the fourth cause of death by cancer among women and the most fatal among gynecological tumors. The purpose of this symposium has been to report and discuss the new developments in the treatment of patients with ovarian cancer, the majority of whom still present with advanced disease. It also tries to make clear to the participants what is evidence-based and what is not. Although the main topic of the symposium is advanced disease, this edition included some updated information on the treatment of early disease under the heading "is early disease really early"; the importance of screening and molecular approaches are highlighted. In addition studies reported in the literature on the role of chemotherapy versus no chemotherapy in high-risk patients with early ovarian cancer have been updated. The pace of new agent development has increased, and it would be helpful to have more efficient preclinical models and early phase-clinical trials to guide the selection of active agents for phase III evaluation. Reaching international consensus is a challenge, but offers the opportunity to test multiple regimens more efficiently against a single control population, rather than conducting multiple smaller studies with redundant internal controls. If indeed answers to the relevant questions are to be obtained more quickly, then a network of current national or international groups could potentially facilitate this. PMID- 11489011 TI - Clarification of conceptualizations of empathy. AB - BACKGROUND: If understanding our clients needs, emotions, and circumstances is fundamental to nursing practice, and empathy is the foundation of that understanding, then a conceptualization of empathy that can be used by nurses is of utmost importance to the profession. In 1992, Morse, Anderson, Bottorff, Yonge, O'Brien, Solberg and McIlveen analysed the concept of empathy in the psychological and nursing literature, and suggested the conceptualization of empathy was incomplete. Since that time, nurse authors have continued to publish conceptualizations and research on empathy. PURPOSE: The purpose of our analysis was to describe empathy as presented in the nursing literature between 1992 and 2000. METHOD: A concept clarification methodology of concept analysis was used because of the many definitions, the rich descriptions, and the application of empathy as a research variable in the reviewed literature. FINDINGS: Five conceptualizations of empathy were revealed: empathy as human trait, empathy as a professional state, empathy as a communication process, empathy as caring, and empathy as a special relationship. CONCLUSIONS: The literature reviewed contained evidence that the concept is developing more depth and breadth. Nurse authors are approaching empathy from a variety of perspectives, time frames, measurements, and outcomes. While all are important to the development of the concept, further enrichment of the conceptual work on empathy is needed before a fully mature concept emerges that is fully useful in nursing practice, research, and education. PMID- 11489012 TI - Concept analysis: abuse of ageing caregivers by elderly care recipients. AB - PURPOSE: The purpose of this article is to clarify the concept of abuse within the context of ageing women who are at risk for or experiencing physical or emotional injury inflicted by elderly family members for whom they provide care. BACKGROUND: The study of abuse of ageing individuals in family caregiving situations has traditionally focused on abuse of the dependent care receiver. However, evidence supports the health risks related to abuse of ageing caregivers as well. Women, usually spouses, daughters, or daughters-in-law, most frequently assume the caregiver role. METHODS: A modification of the strategies for concept analysis proposed by Walker and Avant (1995) is used to clarify the concept of caregiver abuse. Searches of the professional literature reveal that caregiver abuse is rarely addressed; therefore, the broader concept of elder abuse is reviewed and then placed within the general context of family caregiving. Audiotapes of the first session of a community based intervention research study entitled Intervention for the Abuse of Ageing Caregivers (Phillips et al., NIH Grant No. R01 DA-AG11155-01, 1996), in which ageing women caregivers described abusive caregiving situations, were analysed qualitatively using the principles of concept analysis. The audiotapes serve as a second source of data for the concept analysis process. FINDINGS: Antecedents, defining characteristics, and consequences of abuse of ageing caregivers were identified through the process of concept analysis. Model, contrary, and borderline cases are presented to illustrate the findings. CONCLUSIONS: Findings supported the need for awareness that ageing caregivers can be placed at risk by verbally and physically abusive behaviours of the elders for whom they provide care. Use of the term 'abuse' by health care professionals has potentially negative consequences for identification and intervention in cases of potential or actual caregiver abuse. PMID- 11489013 TI - Critical analysis of everyday self-care decision making in chronic illness. AB - AIMS OF THE PAPER: The purpose of the paper is to (1) identify prevalent assumptions that underlie the traditional conceptualization of everyday self-care decision making and (2) contrast these with the findings of relevant research. BACKGROUND/RATIONALE: Current understandings of self-care decision making in chronic illness tend to be extrapolated from knowledge gained in relation to one time decisions, or decision making in contexts that are only superficially related to the complexity and pervasiveness of living with a chronic disease. FINDINGS: The authors challenge the assumptions with which current understandings represent self-care decision making in chronic illness, using evidence from their research on what it is like to live with and manage the implications of having a chronic disease on an everyday basis. CONCLUSIONS: The paper concludes with a call for a new conceptualization of self-care decision making in chronic illness which sufficiently addresses the unique and complex nature of such decisions. PMID- 11489014 TI - Intuition: an important tool in the practice of nursing. AB - AIM: The aim of the study reported here was to evaluate the role of intuition, to examine nurses' understanding of intuition and their perceptions of their use of intuition, and to assess the impact of intuition on nursing practice. BACKGROUND: When we read the dictionary definition of intuition, we have a sense of the meaning but there is also a sense of something still not quite defined. Yet we instinctively know what intuition is. That is, of course, an example of intuition in itself: an understanding of the concept based on our feelings, knowledge and experience. Intuition is a concept neither clearly articulated nor adequately theorized in nursing. There is evidence to suggest that practising nurses use intuition and that its use can change outcomes for patients. Because of its influence on patient well being, it needs to be recognized as an important tool in the practice of nursing. The very nature of intuition determines that there will be little agreement on a precise definition and little empirical evidence to support its existence or worth. Neither of these issues detracts from the fact that intuition is an important part of nursing. METHODS: Grounded theory was used as the overarching theoretical and methodological framework for this study. Using focus group interviews and the Delphi survey technique, data were collected from 262 Registered Nurses who volunteered to take part in the study. FINDINGS: The theory that emerged from this study provides nurses with a way of articulating their understanding of intuition and their perceptions of its use in nursing practice. CONCLUSIONS: Intuition is not some mystical power that appears from nowhere, with no rational explanation or basis. The findings from this study show that it is a product of the synergy that occurs as a result of the interaction of a number of factors. PMID- 11489015 TI - Decision making for health care professionals: use of decision trees within the community mental health setting. AB - AIM OF THE PAPER: To examine the application of the decision tree approach to collaborative clinical decision-making in mental health care in the United Kingdom (UK). BACKGROUND: While this approach to decision-making has been examined in the acute care setting, there is little published evidence of its use in clinical decision-making within the mental health setting. The complexities of dual diagnosis (schizophrenia and substance misuse in this case example) and the varied viewpoints of different professionals often hamper the decision-making process. This paper highlights how the approach was used successfully as a multiprofessional collaborative approach to decision-making in the context of British community mental health care. DESIGN: A selective review of the relevant literature and a case study application of the decision tree framework. CONCLUSIONS: The process of applying the decision tree framework to clinical decision-making in mental health practice can be time consuming and client inclusion within the process is not always appropriate. The approach offers a method of assigning numerical values to support complex multiprofessional decision-making as well as considering underpinning literature to inform the final decision. Use of the decision tree offers a common framework that can assist professionals to examine the options available to them in depth, while considering the complex variables that influence decision-making in collaborative mental health practice. Use of the decision tree warrants further consideration in mental health care in terms of practice and education. PMID- 11489016 TI - Adaptation of an instrument to measure the informational needs of men with prostate cancer. AB - AIM OF THE STUDY: The aim of this study is to adapt an instrument suitable for assessment of the informational needs of men with prostate cancer. BACKGROUND: In recent years prostate cancer has become an important public health problem world wide with considerable social and economic consequences. It is reported that it is the most common cancer affecting British men, with an average lifetime risk of occurrence of one in twelve. DESIGN/METHODS: Methodological research was conducted to develop an instrument to assess the informational needs of men with prostate cancer on hormonal manipulation therapy (HMT) regarding their disease and treatment. The Toronto Informational Needs Questionnaire (TINQ-BC) (Galloway et al. 1997) was modified for use with this client group and was applied to a sample of 90 men generated from three urology centres in Northern Ireland. RESULTS/FINDINGS: Construct and content validity of the instrument was established. Internal consistency reliability using Cronbach's alpha was calculated and found to be satisfactory (0.92). Using confirmatory factor analysis, factor loadings ranging from 0.37 to 0.90 were obtained and considered satisfactory. The subsections of the TINQ-BC categorized as Disease, Investigative tests, Treatment, Psychosocial and Physical needs were confirmed as individual factors. These results indicate that this instrument can be validly applied to this client group. As the instrument was initially developed in Canada and successfully used in the United Kingdom (UK), it is suggested that this instrument also has the potential for cross-cultural application. It has the potential to be used as a clinical reference instrument to assess the informational needs of this patient group. Health care professionals must be aware of the domains of information that these men perceive important so that educational interventions can be accurately and appropriately planned. PMID- 11489017 TI - Expanding the enablement framework and testing an evaluative instrument for diabetes patient education. AB - BACKGROUND: Strategies to delay or prevent complications from diabetes include diabetes patient education. Diabetes educators seek to provide education that meets the needs of clients and influences positive health outcomes. AIMS: (1) To expand prior research exploring an enablement framework for patient education by examining perceptions of patient education by persons with diabetes and (2) to test the mastery of stress instrument (MSI) as a potential evaluative instrument for patient education. METHOD: Triangulated data collection with a convenience sample of adults taking diabetes education classes. Half the sample completed audio-taped semi-structured interviews pre, during and posteducation and all completed the MSI posteducation. Qualitative data were analysed using latent content analysis, descriptive statistics were completed. RESULTS: Qualitative analysis revealed content categories similar to previous work with prenatal participants, supporting the enablement framework. Statistical analyses noted congruence with psychometric findings from development of MSI; secondary qualitative analyses revealed congruency between MSI scores and patient perceptions. CONCLUSIONS: Mastery is an outcome congruent with the enablement framework for patient education across content areas. Mastery of stress instrument may be a instrument for identification of patients who are coping well with diabetes self-management, as well as those who are not and who require further nursing interventions. PMID- 11489018 TI - 'Best research practice': in pursuit of methodological rigour. AB - RATIONALE: This paper is based on the rationale that misuse of methodological notions in research publications lays research studies open to criticism and dismissal. AIM: In search of 'best research practice', this paper aims to examine the different qualities of four major qualitative methodologies: ethnography, descriptive phenomenology, interpretative phenomenology/hermeneutics and critical social theory. DESIGN: The study presents a critical overview of methodological decision-making, illustrating the sorts of issues researchers must consider in order to justify to the readership and to themselves the employment of a particular methodology. This is presented alongside a general overview of qualitative research and a precis of each of the major qualitative methodologies. The paper describes the methodologies, salient features, and examines methodological similarities and differences. The paper concludes by examining the need for methodological rigour within the framework of the National Health Service (NHS) Executive's drive for evidence-based practice in health care. RECOMMENDATIONS: It is hoped that the paper will stimulate a deeper exploration of methodological rigour in future research publications. PMID- 11489019 TI - Response to 'A comparative study of death anxiety in hospice and emergency nurses' by S.A. Payne, S.J. Dean & C. Kalus (1998) Journal of Advanced Nursing 28, 700-706. PMID- 11489020 TI - Response to 'The idea of nursing science' by S.D. Edwards (1999) Journal of Advanced Nursing 29, 563-569. PMID- 11489021 TI - Response to 'Midwives' support needs as childbirth changes' by M. Kirkham & H. Stapleton (2000) Journal of Advanced Nursing 32, 465-472. PMID- 11489022 TI - Response to: Male appropriation and medicalization of childbirth; an historical analysis by H.A. Churchill (2001) Journal of Advanced Nursing 33, 334-342. PMID- 11489024 TI - Evaluating psychosocial nursing interventions for cardiac clients and their caregivers: a case study of the community rehabilitation network in Hong Kong. AB - AIM OF THE STUDY: The purpose of this exploratory study was to describe the type and nature of psychosocial nursing interventions provided for cardiac clients in Hong Kong's Community Rehabilitation Centres (CRNs). DESIGN/METHODS: Using a two phase case study design data were collected from nurses, cardiac clients and their caregivers. The initial phase focused on identifying the types of psychosocial interventions provided. These data were obtained by observing nursing activities and the keeping of daily journals by the nurses. The second phase acquired data from structured telephone interviews, that reviewed clients' perceptions of the psychosocial interventions provided by the nurses and face-to face interviews with clients and their caregivers. FINDINGS: These findings revealed that the nurses' interpersonal skills of information giving, social support and counselling were highly valued by cardiac clients and their caregivers. Both clients and caregivers outlined a concern about the lack of individual care and this was reflected in the focus of the CRN on group processes. CONCLUSION: Recommendations for psychosocial cardiac care are made for public health nurses and nurses working in community settings. PMID- 11489025 TI - Are auditory warnings in the intensive care unit properly adjusted? AB - The purpose of this study was to determine whether auditory warnings in the intensive care unit (ICU) were properly adjusted. An intervention study (before- and-after assessment) was conducted in a 12-bed medical-surgical ICU of an acute care teaching hospital in Barcelona, Spain. A total of 100 patients with stable haemodynamic and respiratory parameters were included. In the first 3-month phase of the study, minimum and maximum alarm parameters of breathing rate, expired volume/min, airway pressure, SaO(2), arterial blood pressure and heart rate were recorded. In the second 12-month phase of the study, the same alarm parameters were recorded every 4 hours in the patient's medical record. In the third 3-month phase of the study, alarm readings were recorded again as in the first phase. The change throughout coefficient of variation (CV) and the 95% confidence interval (CI) for each alarm were calculated. Following the intervention, there was a statistically significant improvement in alarm readings for expired volume, heart rate and systolic blood pressure, so that alarms had been more properly adjusted to the patient's real value. Nursing staff should be aware that auditory warnings in ICU stable patients are frequently set very far from suitable values. Recording of alarm parameters in the patient's medical record as a routine daily activity was an effective intervention for improving adjustment of auditory warnings. PMID- 11489026 TI - The effects of handholding on anxiety in cataract surgery patients under local anaesthesia. AB - AIM OF THE STUDY: To assess the effectiveness of handholding on the anxiety of patients undergoing planned cataract surgery under local anaesthesia. BACKGROUND: Anxiety concerning anaesthesia, pain, physical injuries, isolation, prognosis, possibilities of deformity, or loss of self-control may be stressful to patients undergoing surgery. Anxiety activates the sympathetic nervous system, characterized by an increase in catecholamine concentration, heart rate and blood pressure and increased glucocorticoid levels; it also affects immune responses. Therefore, there is a need for nursing interventions to reduce the anxiety of patients under local anaesthesia who are conscious. METHODS: An untreated control group design with pre and post-test was used. Among 62 patients, 30 were randomly assigned to the handholding group and 32 to the control group. Handholding was provided to subjects of the handholding group during surgery. Visual analogue scales and interviews were used to measure anxiety, and pulse rate and systolic and diastolic blood pressure were used as physiological measures of stress. Blood was taken for analysis of levels of epinephrine, norepinephrine, cortisol, neutrophils, lymphocytes and natural killer cells. RESULTS: The number of subjects who reported decreased anxiety during operation was significantly higher in the handholding group compared with the control group and most of the subjects reported that handholding during operation was very helpful in reducing anxiety. Epinephrine levels in the handholding group were significantly lower than in the control group. CONCLUSIONS: Results suggest that this noninvasive intervention has potential for reducing anxiety in patients having cataract surgery under local anaesthesia. PMID- 11489027 TI - Eating difficulties, complications and nursing interventions during a period of three months after a stroke. AB - AIM: The aim of this study was to describe eating difficulties and especially swallowing in patients with dysphagia, types of nursing intervention, and the development of complications over 3 months. The aim was also to explore common characteristics of eating difficulties that influenced the ability to finish meals. METHODS: Twenty-four consecutive patients admitted because of stroke and dysphagia were included. Nursing interventions, based on assessments, were individually designed. RESULTS: Three subgroups could be identified: those (n=9) who were unable to complete a meal, despite assisted feeding, because of reduced alertness/energy and impaired swallowing function; those (n=5) who could complete a meal, despite suffering from reduced alertness/energy; and those (n=10) who could complete meals with minor difficulties. Patients in the first two groups developed complications such as respiratory infections and/or malnutrition. There was a tendency towards that complications in the third group were less frequent and the hospital stay was significantly shorter than in the other groups. CONCLUSION: The level of alertness/energy in patients with dysphagia after stroke was important for the ability to eat and swallow and the development of complications over time, and thus of great importance for the interventions applied. PMID- 11489028 TI - Feeling at home in nursing homes. AB - AIM: The purpose of this study was to examine determinants of feeling at home and in particular the privacy in nursing homes in The Netherlands. The first question was to what extent nursing homes differed in the degree residents feel at home and experience privacy. The second question was whether feeling at home is related to privacy in the nursing homes. BACKGROUND: Feeling at home is of particular importance to residents of nursing homes because the average length of stay in The Netherlands is about 16 months. A growing number of people are of the opinion that the privacy of the residents has to be improved. Privacy in nursing homes, therefore, has been high on the political agenda over the last few years. Four aspects of privacy were distinguished: privacy related to the characteristics of the building, privacy as a consequence of the attitude of nurses towards residents, privacy in terms of choice and control over time schedules and activities, and privacy related to the amount of disturbance caused by other residents. DESIGN: The data were collected from individual interviews with 686 residents and family-members in 36 nursing homes in The Netherlands. RESULTS: Following quantitative data analysis, it was found that nursing homes differed in the proportion of residents feeling at home as well as in three aspects of privacy. Differences in feeling at home were found to be related to privacy, especially to the perceived attitudes of nurses and perceived disturbance caused by other residents. A weak relationship was found between residents and their perceived privacy of the building. CONCLUSIONS: A significant amount of variation in privacy was found at the nursing home level. This implies that feeling at home is not only individually determined but can be influenced by the nursing home's management policy. PMID- 11489029 TI - A study of the impact of discharge information for surgical patients. AB - AIM OF THE STUDY: To establish whether the routine information surgical patients receive about the management of pain and wound care during their hospitalization is sufficient for them to care for themselves without seeking assistance from a health professional or health care agency. BACKGROUND: While there has been considerable evidence suggesting cost benefits of discharge information the health care environment is constantly changing. Contemporary issues such as increased patient participation, extensive use of technology, reduction in health care expenditure, and greater awareness of consumer rights necessitate further inquiry into the appropriateness of discharge information. METHOD: One hundred and fifty-eight adult patients discharged within a week of their operation participated in the study. A written questionnaire was distributed within 24 hours prior to discharge and a telephone interview conducted 1 to 2 weeks after discharge. At the time of discharge the majority of patients had received information. FINDINGS: Those patients who had received information were less likely to access a health facility than those who had not received information. However, the telephone interview, revealed that there was no evidence that patients who believed they were well informed within 24 hours of discharge about the management of their wound, still felt well informed 1 to 2 weeks later. CONCLUSION: Nurses need to be aware that patients who leave the hospital with little or no discharge information may not be confident in the management of their health condition and therefore may access a health facility, if even just for reassurance. PMID- 11489030 TI - Seniors' decision making about pain management. AB - BACKGROUND: Musculoskeletal pain is a problem with which many seniors must contend, many on a daily basis. Little is known, however, of the self-care decisions that seniors make regarding the management and control of this pain. These decisions can influence in a significant manner the delivery of health care to seniors and their overall health and well-being. Purpose. The purpose of our exploratory-descriptive study was to investigate seniors' decision making regarding the management and control of musculoskeletal pain by gathering data about the context of decision making, the types of decisions seniors made, their decisional conflict and the resources they used in decision making. METHODS: Focus groups and a mail-back questionnaire were used. Data were gathered in 1997 from a convenience sample of 50 seniors in Canada who experienced musculoskeletal pain of a noteworthy nature. RESULTS: Findings revealed that participants made decisions within a context of ageing and the health and social consequences of advancing age. The types of decisions they made included to ignore their pain and to use distraction. They also used exercise, the application of heat and cold, and medications to manage pain. Decisional conflict was minimal and consultation with family and friends superseded that with professionals. CONCLUSIONS: This study contributed to knowledge of decision making in later life about health matters generally, and the management and control of pain specifically. Findings point to the resourcefulness of seniors with respect to self-care and decision making. Seniors want to make informed decisions. However, they need information about the risks and benefits of decisions. PMID- 11489031 TI - Humour between nurse and patient, and among staff: analysis of nurses' diaries. AB - AIM OF THE STUDY: The aim of this study was to investigate the occurrence of humour both between nurses and patients and among nursing staff. BACKGROUND: Humour has positive effects on both a person's physiology and the psyche. Laughter signifies positive things to people; it may add to feelings of togetherness, closeness and friendliness. The use of humour is an indication of mental well-being and it minimizes the effect of negative factors and protects from difficulties. METHODS: The data consisted of diaries written by nurses (n=16), which were analysed inductively using content analysis. FINDINGS: The main categories of humour between nurse and patient were nurse-based humour, co operation-orientated humour and patient-orientated humour. Humour among staff consisted of the main category of resource-orientated humour. CONCLUSION: The consequences of humour between nurse and patient enabled them both to cope with various unpleasant procedures. It helped them to manage difficult situations and led to an improvement in the working climate. Humour among staff helped nurses to cope with their work and created a better atmosphere on the ward. PMID- 11489032 TI - A psychoanalytically informed conversation with a woman and her husband following major surgery for cancer of her neck and torso. AB - AIMS OF THE PAPER: This paper explores psychoanalytic ideas of containment as described by Wilfred Bion and applies them understanding the concerns of a woman with serious illness and her husband. A case vignette shows psychoanalytic thinking in context. The paper explores verbatim exchanges that took place in a room just off a hospital ward and offers a psychoanalytically informed analysis. RATIONALE: This discussion suggests that the impact of physical illness is such that there is potential for the seriously ill to saturate health workers in anxiety and raw feelings. To protect themselves, nurses and other health workers could sometimes unconsciously overlook the emotional care of people in favour of routines, physical aspects of care and other demonstrable methods of treatment. CONCLUSIONS: Drawing on the work of Isabel Menzies Lyth, the discussion concludes by recommending that environments are needed in which a person with serious illness or family member can be helped to make sense of his or her experiences and so integrate them into a meaningful life continuum. Similar conditions are perhaps needed for nurses or other health workers. Appropriate methods of work discussions in clinical supervision might conceivably provide these important requirements. PMID- 11489033 TI - Recent developments in the evidence-based healthcare debate. PMID- 11489034 TI - Meaning and measurement: an inclusive model of evidence in health care. AB - Evidence-based approaches are assuming prominence in many health-care fields. The core ideas of evidence-based health care derive from clinical epidemiology and general internal medicine. The concept of evidence has yet to be analysed systematically; what counts as evidence may vary across disciplines. Furthermore, the contribution of the social sciences, particularly qualitative methodology, has received scant attention. This paper outlines a model of evidence that describes four distinct but related types of evidence: qualitative-personal; qualitative-general; quantitative-general and quantitative-personal. The rationale for these distinctions and the implications of these for a theory of evidence are discussed. PMID- 11489035 TI - Rationality in medical decision making: a review of the literature on doctors' decision-making biases. AB - The objectives of this study were to describe ways in which doctors make suboptimal diagnostic and treatment decisions, and to discuss possible means of alleviating those biases, using a review of past studies from the psychological and medical decision-making literatures. A number of biases can affect the ways in which doctors gather and use evidence in making diagnoses. Biases also exist in how doctors make treatment decisions once a definitive diagnosis has been made. These biases are not peculiar to the medical domain but, rather, are manifestations of suboptimal reasoning to which people are susceptible in general. None the less, they can have potentially grave consequences in medical settings, such as erroneous diagnosis or patient mismanagement. No surefire methods exist for eliminating biases in medical decision making, but there is some evidence that the adoption of an evidence-based medicine approach or the incorporation of formal decision analytic tools can improve the quality of doctors' reasoning. Doctors' reasoning is vulnerable to a number of biases that can lead to errors in diagnosis and treatment, but there are positive signs that means for alleviating some of these biases are available. PMID- 11489036 TI - Introduction to clinical reasoning. PMID- 11489037 TI - Evidence-based medicine and randomized double-blind clinical trials: a study of flawed implementation. AB - The randomized double-blind clinical trial (RDBCT) is a key source of information for evidence-based medicine. However, anomalous and unexplainable results have prompted suggestions that 'unknown and unidentifiable biases' may exist. This paper identifies that a possible flaw in the implementation of RDBCTs may account for these biases. The flaw relates to the breaking of the double blind through the generation of beliefs and expectations in experimenters. These, in turn, may lead to unconscious biases in assessment and cues to patients. It is then uncertain how much of an observed effect is due to such expectations or the treatment itself. Therefore, any RDBCT in which the maintenance of blinding is not monitored throughout its course is at risk of its conclusions being compromised. It is not sufficient to assert that blinding must have been maintained through arguments based on design features. The burden of proof is on the researchers to demonstrate, through data, that blinding has been maintained. The need to address social psychological issues in implementing RDBCTs is discussed and it is recommended that to avoid this confound, methods of monitoring and accounting for experimenter beliefs and expectations should be routinely included in future RDBCTs. PMID- 11489038 TI - Including qualitative research in systematic reviews: opportunities and problems. AB - Qualitative research has been increasingly recognized in recent years as having a distinctive and important contribution to make to health care research. It is capable of being used as a methodologically sufficient approach in its own right, as a precursor to quantitative studies, during or after trials to explain processes and outcomes, and as a means of enhancing the link between evidence and practice. However, qualitative research has been little used as an evidence resource for systematic reviews. We argue that formal synthesis of both qualitative and quantitative forms of research is essential, and we discuss some of the problems that need to be overcome in carrying out such syntheses. These include methodological prejudice, problems in searching for qualitative evidence, and issues in synthesizing qualitative data. We call for progress to be made on the science and methods of including qualitative research in the evidence base of medicine. PMID- 11489039 TI - An illustrated guide to the methods of meta-analysis. AB - Meta-analysis is now accepted as a necessary tool for the evaluation of health care. Such analyses have been carried out in virtually every area of medicine to evaluate a wide spectrum of health care interventions and policies. This paper has three broad aims: (1) to describe the basic principles of meta-analysis, using a meta-analysis of interventions intended to reduce hospital re-admission rates for illustration; (2) to consider threats to the internal validity of meta analysis, and the measures which can be taken to minimize their impact; and (3) to present an overview of more specialist and developing methods for synthesizing data, with the intention of outlining the directions meta-analysis may take in the future. The methods used to synthesize studies, which take 'weighted averages' of effect sizes have been refined to a high degree, while the methods for dealing with threats to the validity of meta-analyses such as publication bias, and variations in quality of the primary studies, are at a less advanced stage. However, many consider this standard 'weighted average' approach to meta analysis not to be 'state of the art' in at least some situations, where the use of more sophisticated methods, generally to explain variation in estimates from different studies and synthesize a broader base of evidence, would be advantageous. Currently, approaches which attempt to do this are mainly still in the experimental stage and, unfortunately, ideas which sound natural and appealing are often difficult to implement in practice. Clearly, it will be some time before they are used routinely, but significant steps have been made. PMID- 11489040 TI - A critique of hypertension treatment trials and of their evaluation. AB - Progressive improvements in antihypertensive drug therapy over the past four decades have provided clear benefits in limiting cardiovascular complications. Unfortunately, and largely inexplicably, the inclusion in meta-analyses of defective trials plus the employment of inappropriate diagnostic criteria for adverse coronary events have led to spurious, exaggerated claims for the advantages of such treatment. The consequence has been a devaluation of the very real worth of prophylactic drug therapy for hypertension. PMID- 11489041 TI - The potential influence of small group processes on guideline development. AB - The authors discuss the important and often neglected role of psychosocial factors in guideline development. Such factors can influence the process by which guideline development groups interact, make decisions and achieve consensus, which may have important implications for the validity and reliability of the recommendations they produce. Particular attention is paid to the role of social influence within multidisciplinary groups and the importance of good chairmanship. Recommendations for participants, chairmen and guideline development bodies are given. PMID- 11489042 TI - Publishing web-based guidelines using interactive decision models. AB - Commonly used methods for guideline development and dissemination do not enable developers to tailor guidelines systematically to specific patient populations and update guidelines easily. We developed a web-based system, ALCHEMIST, that uses decision models and automatically creates evidence-based guidelines that can be disseminated, tailored and updated over the web. Our objective was to demonstrate the use of this system with clinical scenarios that provide challenges for guideline development. We used the ALCHEMIST system to develop guidelines for three clinical scenarios: (1) Chlamydia screening for adolescent women, (2) antiarrhythmic therapy for the prevention of sudden cardiac death; and (3) genetic testing for the BRCA breast-cancer mutation. ALCHEMIST uses information extracted directly from the decision model, combined with the additional information from the author of the decision model, to generate global guidelines. ALCHEMIST generated electronic web-based guidelines for each of the three scenarios. Using ALCHEMIST, we demonstrate that tailoring a guideline for a population at high-risk for Chlamydia changes the recommended policy for control of Chlamydia from contact tracing of reported cases to a population-based screening programme. We used ALCHEMIST to incorporate new evidence about the effectiveness of implantable cardioverter defibrillators (ICD) and demonstrate that the cost-effectiveness of use of ICDs improves from $74 400 per quality adjusted life year (QALY) gained to $34 500 per QALY gained. Finally, we demonstrate how a clinician could use ALCHEMIST to incorporate a woman's utilities for relevant health states and thereby develop patient-specific recommendations for BRCA testing; the patient-specific recommendation improved quality-adjusted life expectancy by 37 days. The ALCHEMIST system enables guideline developers to publish both a guideline and an interactive decision model on the web. This web-based tool enables guideline developers to tailor guidelines systematically, to update guidelines easily, and to make the underlying evidence and analysis transparent for users. PMID- 11489043 TI - Clinical practice guidelines: when the tool becomes the rule. AB - This work provides a critical examination of the use of clinical practice guidelines to measure individual performance. The problems inherent in using a measure of central tendency derived from a distribution of individual performances are addressed, as is the translation of the collectively determined guidelines into a measurement instrument. It is suggested that every process on the distribution of processes used to determine the guideline must be considered equally legitimate representations of the process in question. It is further suggested that to accept as a standard of quality, a particular process simply because there is a minimum of variation between providers, is to ignore the importance of the linkage between process and outcome. The importance of an independent measure of quality based on outcomes is further emphasized by highlighting the tautological nature of analyses that include an input measure, such as nursing hours, in both the dependent variable and the list of independent variables. It is recommended that individual performance be evaluated within the tolerances of the distribution from which they were derived and not be held to some measure of central tendency of that distribution. The alternative is to use the measure of central tendency with plus or minus limits such as one, or more, standard deviations. PMID- 11489044 TI - Evidence-based medicine in general practice: beliefs and barriers among Australian GPs. AB - If implemented, evidence-based medicine (EBM) in general practice will improve health outcomes for patients. This paper examines the views of 60 Australian general practitioners about EBM. While 57% of respondents had a computer in their surgery, 15% had Internet access and only 3% had access to the Cochrane Library at work. The most commonly cited barrier to EBM was 'patient demand for treatment despite lack of evidence for effectiveness' (45%). The next three most highly rated barriers related to lack of time. For each of three tasks of EBM, namely searching for evidence, appraising evidence and discussing the implications of evidence with patients, lack of time was rated as a 'very important barrier' by significantly more participants than lack of skills (McNemar's tests: chi2(1) = 7.1, P = 0.008, chi2(1) = 14.0, P = 0.001 and chi2(1) = 9.0, P = 0.003, respectively). Preferred resources for EBM included clinical practice guidelines (rated as 'very useful' by 55%) and journals that summarize research evidence, for example Evidence-based Medicine (52%). Systematic reviews were considered 'very useful' by only 15% of respondents, consistent with our finding that 30% did not understand the term 'systematic review'. Furthermore, 43% did not understand 'meta-analysis'. A minority indicated they understood the terms 'relative risk' (23%), 'absolute risk' (28%) and 'number needed to treat' (15%) sufficiently to explain to others. Skills development is crucial to achieve EBM in general practice. PMID- 11489045 TI - Shared decision making observed in clinical practice: visual displays of communication sequence and patterns. AB - The aim of the study was to examine the communication strategies of general practitioners attempting to involve patients in treatment or management decisions. This empirical data was then compared with theoretical 'competences' derived for 'shared decision making'. The subjects were four general practitioners, who taped conducted consultations with the specific intent of involving patients in the decision-making process. The consultations were transcribed, coded into skill categorizations and presented as visual display using a specifically devised sequential banding METHOD: The empirical data from these purposively selected consultation from clinicians who are experienced in shared decision making did not match suggested theoretical frameworks. The views of patients about treatment possibilities and their preferred role in decision making were not explored. The interactions were initiated by a problem-defining phase, statements of 'equipoise' consistently appeared and the portrayal of option information was often intermingled with opportunities to allow patients to question and reflect. A decision-making stage occurred consistently after approximately 80% of the total consultation duration and arrangements were consistently made for follow-up and review. Eight of the 10 consultations took more than 11 min - these specific consultations were characterized by significant proportions of time provided for information exchange and patient interaction. The results demonstrate that some theoretical competences are not distinguishable in practice and other stages, not previously described, such as the 'portrayal of equipoise', are observed. The suggested ideal of a shared decision-making interaction will either require more time than currently allocated, or alternative strategies to enable information exchange outside the consultation. PMID- 11489046 TI - Effectiveness of educational interventions on the improvement of drug prescription in primary care: a critical literature review. AB - This paper is a critical review of studies of educational programmes designed to improve prescription practices in ambulatory care. Scientific articles were selected from the following bibliographical indices: MEDLINE, IME, ICYT and ERIC. The searches covered the time period between 1988 and 1997. The search criteria included: primary-care, educat*, prescription* and other related keywords. The inclusion criteria were studies describing educational strategies aimed at general practitioners working in ambulatory settings. The study outcome was change in prescribing behaviour of physicians through prescribing indicators. The following data were extracted: study design, target drugs, type of intervention, follow-up period of the prescription trends, type of data analysis, type of statistical analysis and reported results. We found 3233 articles that met the search criteria. Of these, 51 met the inclusion criteria and 43 studied the efficacy/effectiveness of one or various interventions as compared to no intervention. Among seven studies evaluating active strategies, four reported positive results (57%), as opposed to three of the eight studies assessing passive strategies (38%). Among the 28 studies that tested reinforced active strategies, 16 reported positive results for all variables (57%). Eight studies were classified as a high degree of evidence (16%). We concluded that the results of our review suggest that the more personalized, the more effective the strategies are. We observe that combining active and passive strategies results in a decrease of the failure rate. Finally, better studies are still needed to enhance the efficacy and efficiency of prescribing practices. PMID- 11489047 TI - Exploring the pathology of quality failings: measuring quality is not the problem -changing it is. AB - The USA can boast a long history of investigation into quality failings in health care. From Ernest Codman and Abraham Flexner in the opening decades of this century through to the intense activity of the 1980s and 1990s, much careful study has exposed extraordinary and at times scandalous deficiencies in the quality of care (Millenson 1997; Chassin & Galvin 1998; Schuster et al. 1998). Yet we are still far from developing 'industrial strength' quality in health care: in all but a few isolated areas, such as general anaesthesia, 'six sigma quality' (i.e. a handful of errors per million) seems wishful thinking (Chassin 1998). Pockets of excellence and innovation notwithstanding, the dominant experience of the past two decades has been an increasing ability to document quality failings and a seeming inability to mobilize effective action (Coye & Detmer 1998). The rich literature on health-care quality that has sprung up over the past few decades has largely failed to provide a clear direction for quality improvement activity. This paper analyses some of the reasons why this might be so. Contrasting the relative absence of progress on health-care quality with the relative success of disease epidemiology provides some illuminating parallels. In essence, study of the quality of care has focused largely on providing a 'descriptive epidemiology'. Much more work is needed yet to unravel the underlying pathology of quality failings, in order to empower development of an 'aetiological epidemiology' of quality in health care. Such understanding is essential as a precursor to targeted and effective preventative and remedial action. PMID- 11489048 TI - Errors: can indicators measure the magnitude? AB - Errors in medicine, especially medication errors, have long been recognized as a dimension of quality of care and organizational performance. Recently, however, the magnitude of the issue, or its potential impact on cost, quality of care and patient safety have catapulted this issue to the forefront of national debate on the appropriateness of patient care management. There are still fundamental issues associated with the measurement of errors. Should errors that do not cause patient harm receive much attention? Could there be organizational or system issues that predispose to errors? Are there acceptable measurement models that allow comparative analysis and trending of institutional error rate profiles? This paper presents a systematic review of the measurement aspects for errors in medicine, emphasizing the medication errors' dimension. An indicator-based, epidemiological model of measurement is proposed which will allow a systematic inquiry into the issues of both preventable and non-preventable errors and their potential for patient harm. PMID- 11489049 TI - Factors associated with expressed satisfaction among people with intellectual disability receiving residential supports. AB - The aim of the present study was to identify factors associated with variations in the levels of expressed satisfaction among adults with intellectual disability (ID) receiving residential supports. Semi-structured interviews were conducted with 96 people with ID. Forty-five subjects lived in village communities and 51 received community-based residential supports. Ratings were made of the participants' expressed levels of satisfaction in seven domains: (1) their home; (2) daytime activities; (3) social and recreational activities; (4) support from services; (5) friendships and relationships; (6) choices available to them; and (7) risks. The data indicated that: (1) interviewees living in village communities expressed greater satisfaction with friendships and relationships than interviewees living in community-based residential supports; (2) in the other six domains of life satisfaction which were investigated, there were no statistically significant differences between groups; (3) interviewees expressed greater satisfaction with their accommodation and day activities than with friendships, risks and support received; and (4) a wide range of variables relating to the personal characteristics of the interviewees and support received were associated with variations in levels of expressed satisfaction. Variation in the levels of expressed satisfaction was reliably associated with variables relating to the personal characteristics of the interviewees and the nature of the support received. PMID- 11489050 TI - Abilities underlying decoding differences in children with intellectual disability. AB - Researchers in recent years have made much progress towards understanding why some children struggle to learn to read. However, little of this research has involved children with intellectual disability associated with an IQ < 70 (ID, also called mental retardation). In the present analysis, the authors examined cognitive similarities and differences between stronger and weaker decoders, all of whom have ID. The 65 children with ID in the present analysis were initially referred by their teachers for a study that involved training basic phonological reading skills. The present analysis compares 21 children who were excluded from the training study because their decoding skills were already too high with 44 children whose decoding skills were low enough for the training study. The groups were compared on general intelligence, language ability, phonemic awareness and phonological memory. Initial analyses showed that the stronger decoders were significantly better than weaker decoders in language ability, phonemic awareness and rehearsal in phonological memory, but not in intelligence. They were also significantly older than weaker decoders. When age was covaried out, the groups differed significantly only in rehearsal in phonological memory, although the difference for phonemic awareness was marginally significant when the poorest performers were excluded. When intelligence is substantially limited, the ability to rehearse or refresh phonological codes in working memory plays a major role in determining children's success in learning to read. This ability appears to be more important than intelligence, language ability and phonemic awareness. It is possible that the reason the phonemic awareness measure was not as good at distinguishing the groups as the phonological rehearsal measure was because the former did not involve assembling phonological output. It is suggested that it is the combination of poor phonological representation and poor phonological output assembly that makes decoding difficult for some children with ID. PMID- 11489051 TI - Cancer incidence among people with intellectual disability. AB - The aim of the present study was to address the unresolved question of the risk of neoplasms among people with intellectual disability (ID). A total of 2173 individuals with ID from a large, representative, nation-wide population study conducted in Finland in 1962 were followed-up for cancer incidence between 1967 and 1997. Standardized incidence ratios (SIRs) were defined as ratios of observed to expected numbers of cancer cases. Expected rates were based on national incidence rates. The observed number of cancers in the cohort (173) was close to what was expected [SIR = 0.9, 95% confidence interval (95% CI) = 0.8-1.0]. There was a significantly reduced risk of cancers of the prostate (SIR = 0.2, 95% CI = 0.0-0.5), urinary tract (SIR = 0.3, 95% CI = 0.1-0.7) and lung (SIR = 0.6, 95% CI = 0.4-1.0). The risk was increased in cancers of the gallbladder (SIR = 2.8, 95% CI = 1.1-5.8) and thyroid gland (SIR = 2.1, 95% CI = 1.0-4.8). The risks of lung and gallbladder cancer were lowest and highest, respectively, in those subjects with profound and severe ID, a group who also had significantly elevated SIRs for brain cancer (SIR = 3.46, 95% CI = 1.5-14.4) and testicular cancer (SIR = 9.9, 95% CI = 1.2-35.6). The incidence of cancer among people with ID was comparable with the general population, despite their low prevalence of smoking and apparently decreased diagnostic screening activity. Nevertheless, a few types of cancer carry a higher risk in the population with ID, possibly because of conditions typical among this group, such as gallstones or oesophageal reflux. PMID- 11489052 TI - Speech disorder in children with Down's syndrome. AB - The speech of children with Down's syndrome (DS) is often unintelligible, unlike many other children who have an intellectual disability. However, the nature of their speech disorder is controversial. The speech error patterns of children with DS were compared to those of intellectually average children with phonological disorder whose errors were characterized by inconsistency. The groups were matched for percentages of consonants produced in error. The data revealed no differences between the groups in terms of the number of words which were produced inconsistently on repeated productions in a picture-naming task. However, further analyses revealed differences in the type of errors made by the groups in that the children with phonological disorder characterized by inconsistent errors made more changes to words on repeated production than the group with DS. The deficits underlying inconsistent pronunciation of words in the two groups of children under investigation would appear to differ. Intervention strategies should target the deficits identified. PMID- 11489053 TI - Kinetic form discrimination in Prader-Willi syndrome. AB - Discrimination of the shape of motion-produced forms generated by random elements (i.e. second-order stimuli varying in element density and temporal correlation) was tested in four groups: (1) subjects with Prader-Willi syndrome (PWS), chromosome 15q deletion subtype; (2) subjects with PWS, uniparental maternal disomy (UPD) subtype; (3) equivalent non-PWS controls; and (4) normal controls. The performance of the normal controls exceeded that of all other groups (78% correct, P < 0.009). The PWS deletion (66%) and the equivalent control groups (59%) did not differ (P < 0.95). The UPD group performed significantly less well (38%, P < 0.04) than all the other groups. The performance of the PWS deletion and equivalent control groups is consistent with other data indicating that these populations encounter difficulty meeting the processing demands posed by second order stimuli. The inferior performance of the UPD group may be attributed to receiving two active alleles of a maternally expressed gene influencing neural development. One candidate is the ubiquitin protein ligase gene (UBE3A), which is maternally expressed only and localized to the 15q region. Other possibilities include the requirement of a paternally expressed gene, residual mosaic trisomy 15 in the brain tissue or complex interactions including specific ratios of differentially spliced gene products. PMID- 11489054 TI - Causal explanations, concern and optimism regarding self-injurious behaviour displayed by individuals with Cornelia de Lange syndrome: the parents' perspective. AB - Research into behaviours associated with specific syndromes, such as Cornelia de Lange syndrome (CdLS), has neglected to explore the parents' perspective, particularly the potential impact of the notion of behavioural phenotypes on parents' causal explanations. Given the research focus on self-injurious behaviour (SIB) in CdLS, the present study examined parental concern across four topographies of challenging behaviour, causal explanations for these behaviours and optimism for change. As part of a larger study, a questionnaire survey of 86 parents of children and adults with CdLS was conducted. Quantitative data on parental concern and optimism with regard to behaviour problems were collected. Causal explanations for behaviour problems were examined by subjecting open-ended responses to a content analysis. Parents were as concerned about physical aggression and disruptive behaviours as they were about SIB. The majority of parents had deconstructed how CdLS might be associated with SIB in terms of other factors associated with CdLS, such as degree of intellectual disability. Parents did not believe that CdLS influenced SIB more than other challenging behaviours and their beliefs did not effect optimism regarding future change in the behaviour. Despite the focus of research on SIB in CdLS, parents of children and adults with CdLS are also concerned about other challenging behaviours. There was no evidence that a deterministic perspective had been adopted by parents and causal explanations were unrelated to optimism for future change. PMID- 11489055 TI - Assessing the need for reactive behaviour management strategies in children with intellectual disability and severe challenging behaviour. AB - The need for reactive behavioural management strategies for aggressive behaviours in adults with intellectual disability has been clearly established, but equivalent information concerning children with challenging behaviour is lacking by comparison. The present study was conducted in order to assess this need. A retrospective study was conducted to ascertain the nature of aggressive behaviours amongst a cohort children referred to a specialist support service. Aggressive behaviours were found to occur at high rates within the study group. Almost 60% of the children displayed aggression that occurred at least daily, and the behaviours resulted in serious consequences for carers in almost one-third of the group. Physical interventions were already in use in 56% of cases, but were largely improvised by carers. The need to include reactive behavioural training as part of an overall intervention package for carers of children with challenging behaviour appeared to be supported by the present results. The implications for training design are discussed. PMID- 11489056 TI - Police attitudes toward people with intellectual disability: an evaluation of awareness training. AB - It is argued that more favourable police attitudes to people with intellectual disability (ID) are essential in meeting the police code of ethics, which stresses impartiality and respect for human dignity. The need to acknowledge and investigate the extent of support for eugenic attitudes in other key professionals who have a significant role in the successful inclusion of people with ID in community settings is discussed. The present paper reports on the evaluation of an awareness training event conducted by the Royal Ulster Constabulary in terms of the impact on attitudes towards people with ID held by police officers. The quasi-experimental design involved the measurement of participants' attitudes prior to and following awareness training, and the comparison of these data with a control group of participants who did not undertake awareness exercises. The Attitudes toward Mental Retardation and Eugenics (AMRE) scale was the instrument used to measure attitudes. Analysis identified the presence of varying degrees of support for the application of eugenic principles to people with ID. Furthermore, the results indicate that participation in the awareness exercise and subsequent discussions is associated with a significant reduction in support for eugenic-based attitudes towards people with ID by the police officers involved. Investment in training events which target attitudes towards people with ID can bring about a shift in reported attitudes. The importance of evaluating such awareness-raising exercises and their impact on police behaviour is highlighted. PMID- 11489057 TI - Effect of unexpected outcomes and causal attributions on key workers' expectancies of goal attainment. AB - Goal planning is an integral part of the role of the key worker in services for people with intellectual disability. The present study investigated the effect of the Weiner stability-expectancy principle on unexpected outcomes in goal planning. Four hundred and twenty-two actual goal attainment scales were constructed, with each scale describing the key workers' expected levels of outcome. Two hundred and twenty-six (54%) unexpected outcomes, both successful and unsuccessful, were then rated by key workers to provide a causal explanation for each outcome. These explanations were then grouped according to whether the attributions represented stable or variable causes. Finally, key workers were asked to provide a new expectancy rating to forecast future goal attainment. Two predictions from Weiner's model were confirmed by statistical analysis: (1) that success attributed to stable factors would lead to higher revised forecasts of goal attainment than success attributed to variable factors; and (2) that failure attributed to stable factors would lead to lower revised forecasts of goal attainment than failure attributed to variable factors. The results of the present study confirm that stability attributions do alter key workers' revised forecasts of goal attainment. The results are discussed in terms of the possible consequences for key worker behaviour, such as goal abandonment, differential treatment of service users and the need for a knowledgeable approach to performance appraisal by managers. PMID- 11489058 TI - Transfer and interference of motor skills in people with intellectual disability. AB - Atypical laterality (i.e. the lack of a clear pattern of lateralization) has been found to be a characteristic feature of individuals with intellectual disability (ID). The evidence for this has been based on 'handedness' studies which have contained little information about the ability of people with ID to carry out interhemispheric tasks reflecting bilateral transfer or interference. The present study examined this capacity in individuals with ID by utilizing bilateral transfer and interference paradigms. Right-handed subjects with ID (IQ = 55-76) and controls matched for age and sex were tested for bilateral transfer of motor skill in contralateral hands with a mirror-drawing task. The subjects were also tested for their ability to perform a finger-tapping task while processing verbal and non-verbal stimuli. The findings indicated that people with ID are significantly deficient relative to matched controls in bilateral transfer of motor skills from their non-preferred (left) hand to their preferred (right) one. The effect of interference during performance of the dual task was significantly greater in individuals with ID. Subjects with ID were found to perform better with their non-preferred than with their preferred hand. A within-group comparison revealed that right-handed performance was more affected by interference than left in these subjects. PMID- 11489059 TI - Chemokines in pathology and medicine. AB - About 50 human chemokines and nearly 20 receptors have been identified and characterized in little more than a decade since the discovery of interleukin 8 (IL-8), the first chemotactic cytokine. Research in this field has dramatically changed our understanding of leucocyte traffic in inflammation and immunity. This paper has been written for scientists and practitioners in the field of medicine. It reviews in concise and intelligible form information that I consider useful for understanding the role of chemokines in human pathophysiology. The main areas covered are: (i) the basics of chemokine structures, mode of action, activities and selectivity; (ii) newer aspects of the broad involvement of chemokines in the regulation of immune defence and the housekeeping of the immune system; (iii) the role of chemokines in pathology as illustrated by animal models and studies of human diseases; and (iv) novel therapeutic approaches for a variety of inflammatory conditions, which are based on modulation of chemokine activity. PMID- 11489060 TI - The dysmetabolic syndrome. AB - The first unifying definition for the metabolic syndrome was proposed by WHO in 1998. In accordance to this, patients with type 2 diabetes mellitus or impaired glucose tolerance have the syndrome if they fulfil two of the criteria: hypertension, dyslipidaemia, obesity/abdominal obesity and microalbuminuria. Persons with normal glucose tolerance (NGT) should also be insulin resistant. About 40% of persons with impaired glucose tolerance (IGT) and 70% of patients with type 2 diabetes have features of the syndrome. Importantly, presence of the dysmetabolic syndrome is associated with reduced survival, particularly because of increased cardiovascular mortality. The dysmetabolic syndrome most likely results from interplay between several genes and an affluent environment. Compatible with the thrifty gene theory, common variants in genes regulating lipolysis, thermogenesis and glucose uptake in skeletal muscle account for a large part of such thrifty genes. However, hitherto unknown genes may still be identified by random gene approaches. PMID- 11489061 TI - The effect of 24 h blood pressure levels on early neurological recovery after stroke. AB - OBJECTIVES: To assess the influence of 24 h blood pressure (BP) levels on functional recovery 1 week after stroke and the effect of antihypertensive therapy on 24 h BP levels. DESIGN: Prospective study of patients admitted to hospital over 1 year with first in a lifetime stroke who underwent 24 h BP and casual measurements. Setting. Medical wards in a teaching hospital. Subjects. Of 160 patients, 72 patients admitted to hospital within 24 h of stroke onset were investigated. Patients with conditions and therapy that interfered with autonomic and sympathetic function were excluded. Interventions. All subjects underwent 24 h BP and casual recordings on admission to hospital and at day seven after stroke. The mean 24 h, day and night systolic BP (SBP) and diastolic BP (DBP) and their differences (nocturnal BP dip) were recorded. Patients were divided into three groups according to whether they were taking antihypertensive therapy during the first week: (i) no therapy, (ii) therapy continued after stroke, and (iii) new therapy introduced. Main outcome measures. Functional recovery (Rankin Scale 0-1) and neurological improvement [Scandinavian Stroke Scale (SSS) >/=3 points] by 1 week of stroke. Change in circadian 24 h BP over 1 week. RESULTS: For each 10 mmHg difference between day and night time DBP, the odds for making a complete recovery were 4.63 (95% CI: 1.57-13.7, P=0.01). For each 10 mmHg difference between day and night SBP, the odds for making an improvement in neurological status was 2.24 (95% CI: 1.16-4.32; P=0.016). Significant falls in 24 h DBP (P=0.01), daytime SBP (P=0.005) and mean arterial BP (MABP) (P=0.04) over 1 week were demonstrated in patients who had just commenced antihypertensive therapy (P=0.001). CONCLUSION: An increase in day to night time BP change is favourable in short-term outcome after acute stroke. Significant falls in BP are more likely in patients started on antihypertensive therapy for the first time. Further research is required to understand the effects of circadian BP rhythm on stroke outcome. PMID- 11489062 TI - Are patients on long-term oxygen therapy followed up properly? Data from the Danish Oxygen Register. AB - OBJECTIVES: The aim of the study was to assess the extent and quality of follow up of patients on LTOT. SETTING: The Danish Oxygen Register. SUBJECTS: A total of 890 chronic obstructive pulmonary disease (COPD) patients who were on long-term oxygen therapy (LTOT) during the period from 1 November 1994 to 31 August 1995. MAIN OUTCOME MEASURES: The extent and quality of follow-up. RESULTS: Only 38.5% of the patients were followed up in the study period, and only 17.5% had a 'sufficient follow-up' defined as at least one follow-up visit within 10 months which included measurement of arterial blood gases or pulsoximetry with oxygen supply, verification that the patient used oxygen > or =15 h day-1 and was nonsmoker. Female gender, LTOT initiated 3-12 months ago, LTOT started by a chest physician at pulmonary department and LTOT prescribed > or =15 h day-1 were found to be significant predictors of 'sufficient follow-up' (odds ratio (OR): 1.7, 2.0, 3.7 and 1.9, respectively). CONCLUSIONS: The extent and the quality of follow-up of patients on LTOT were poor, especially if a nonpulmonary physician initiated LTOT. We recommend that more attention should be paid on proper monitoring of LTOT, and that only chest physicians should be able to prescribe and re-evaluate LTOT. PMID- 11489063 TI - Clinical outcome of patients with suspected pulmonary embolism. A follow-up study of 588 consecutive patients. AB - OBJECTIVE: To investigate the clinical outcome in patients with clinically suspected pulmonary embolism (PE). Design and setting. In a retrospective design we studied 588 consecutive patients with suspected PE and referred for lung scintigraphy from 1995 to 1998. The mean follow-up time was 653 +/- 424 days. RESULTS: The diagnosis of PE was confirmed in 194 and excluded in 394 patients, respectively. The overall prevalence of PE was 33%. Amongst clinical and paraclinical variables, age, chronic obstructive pulmonary disease (COPD), heart rate, pleuritic pain, presence of deep venous thrombosis (DVT), electrocardiographic signs of right ventricular (RV) strain were identified as independent predictors of the diagnosis of PE. Amongst patients with PE anticoagulation was given in 96% for at least 3 months and 13% received thrombolytic therapy. Recurrent PE was seen in 6% of patients with PE whereas none of the patients with no diagnosis of PE suffered PE during follow-up. The 1 year mortality was 18% amongst patients with PE and 15% in patients with excluded PE (P=NS). The cause of death amongst patients with PE was cancer (49%) and PE (28%), whereas patients without PE had an excess mortality because of cancer, COPD, acute myocardial infarction and heart failure. CONCLUSION: Patients admitted to hospital on suspicion of PE have increased risk of adverse clinical outcome whether the diagnosis of PE is confirmed or not. This indicates that the patients where the diagnosis is excluded often suffer from other serious illness that warrants further investigations. PMID- 11489064 TI - Effects of thiazide- and loop-diuretics, alone or in combination, on calcitropic hormones and biochemical bone markers: a randomized controlled study. AB - OBJECTIVE: Diuretics are commonly used drugs that in addition to their effect on the cardiovascular system also affect calcium homeostasis and bone metabolism. We evaluated the effects of loop diuretics (LD) and thiazide diuretics (TD) on calcitropic hormones and biochemical bone markers. DESIGN: A total of 50 postmenopausal women were randomized to 7 days of treatment with either the TD bendroflumethiazide, the LD bumetanide, bendroflumethiazide plus bumetanide, or placebo. Blood and urine (24 h) were sampled on each day. Statistical inferences were made versus the concomitant changes in the placebo group. RESULTS: Bendroflumethiazide increased the tubular reabsorption of calcium (TRCa) (+0.46 +/- 0.11%, P=0.009), plasma levels of parathyroid hormones (PTH) (+24 +/- 10%, P=0.06), and 1,25(OH)2D (+12 +/- 6%, P=0.03). Bumetanide decreased the TRCa (-0.5 +/- 0.1%, P=0.01) and increased plasma PTH and 1,25(OH)2D levels (+27 +/- 9%, P=0.02 and +36 +/- 12%, P=0.006, respectively). Treatment with either of the drugs did not alter plasma calcium, osteocalcin, bone alkaline phosphatase (bone ALP) or urinary NTx/creatinine ratio. However, treatment with both drugs caused an increased plasma calcium level (+2.7 +/- 1.0%, P=0.007) and decreased plasma levels of bone-ALP (-21 +/- 3%, P=0.001), osteocalcin (-6 +/- 3%, P=0.03), and urinary NTx/creatinine ratio (-39 +/- 6%, P=0.001). CONCLUSION: Calcium homeostasis and bone metabolism are to a major degree influenced by diuretic treatment. Surprisingly, LD and TD exerted a similar effect on calcitropic hormones despite their opposite effects on the renal calcium excretion. In clinical practice, treatment with diuretics has to be considered as a cause of parathyroid stimulation. PMID- 11489066 TI - Use of dietary supplements and natural remedies increased dramatically during the 1990s. AB - OBJECTIVES: To estimate the prevalence and trends in dietary supplement and natural remedy use in Sweden during the 1980s and 1990s. DESIGN: Three nationally representative, cross-sectional surveys conducted in 1980-81, 1988-89 and 1996-97 were used for analysis. In face-to-face interviews participants reported consumption of dietary supplements and natural remedies during the previous 2 weeks. SETTING: Sweden. SUBJECTS: The samples consisted totally of 38 594 adults aged 16-84 years (14 642 in the 1980-81 survey, 12 391 in the 1988-89 survey and 11 561 in the 1996-97 survey). Main outcome measures. Changes in prevalence of dietary supplement and natural remedy users between 1980 and 1997. RESULTS: The 70% increase in the prevalence of dietary supplement users amongst both men and women [odds ratio (OR), 1.7; 95% confidence interval (CI), 1.6-1.9, OR, 1.7; 95% CI, 1.5-1.8, respectively] occurred mainly between 1988-89 and 1996-97. The increase in the prevalence of natural remedy users was even more dramatic - more than threefold in men (OR, 3.3; 95% CI, 2.9-4.0) and almost threefold in women (OR, 2.6; 95% CI, 2.3-2.9) - and the systematic increase started already in the 1980s. The increase was observed in all age groups and in all socio-economic groups, except for farmers. In 1996-97 the prevalence of dietary supplement users was 22% amongst men and 33% amongst women, and of natural remedies 7 and 14%, respectively. CONCLUSIONS: During the last two decades, the use of dietary supplements and natural remedies amongst the adult Swedish population has dramatically increased. PMID- 11489065 TI - Consequences of late referral of patients with end-stage renal disease. AB - OBJECTIVES: The aim of the present retrospective single centre study of patients entering renal replacement therapy (RRT), was to evaluate the effects of different referral patterns on morbidity, choice of therapy, and duration of hospitalization in patients with chronic renal failure. SUBJECTS: A total of 242 patients with chronic renal failure starting their first RRT between 1984 and 1998, were divided into three groups. Group 1 (n=80): RRT started 1984-88, group 2 (n=73): RRT started 1989-93 and group 3 (n=89): RRT started 1994-98. Patients were classified as early referrals (ER) or late referrals (LR) depending on whether they started first RRT more than or less than 3 months after first referral to a nephrologist. RESULTS: The proportion of LR was 27.3% (21 patients) in group 1, 27.4% (20 patients) in group 2 and 28.1% (25 patients) in group 3. In the ER, 35 patients (14.5%) received a predialytic kidney transplant, none in the LR. Comparing clinical details, the LR's in group 3 were significantly older than ER [median age 72 (53-81) vs. 56 (15-81) years, P < 0.0001], had a lower serum albumin [median 33.0 (19.0-42.0) vs. 39 (19.0-48.0) g L-1, P < 0.0001], and serum calcium [median 2.0 (1.4-2.6) vs. 2.3 (1.8-2.7) mmol L-1, P < 0.0001]. The ER had a significantly higher use of antihypertensive drugs, calcitriol, phosphate binders, and bicarbonate. Of the patients starting RRT on haemodialysis, all LR started on a temporary vascular access. About 43% of the ER started on a functioning arteriovenous fistula (P < 0.0001). The duration of hospital stay in connection with start of dialysis was 31 days (7-73) in the LR as compared with 7 (1-59) days in the ER (P < 0.0001). CONCLUSIONS: We conclude that in our centre, early referral to nephrologist is associated with lower age, a higher likelihood of predialytic transplantation, better metabolic status at start of RRT, a higher proportion starting haemodialysis on a functioning arteriovenous fistula, and a shorter duration of the initial hospital stay. Further research on health care delivery is warranted. PMID- 11489067 TI - Intraoperative haemorrhage associated with the use of extract of Saw Palmetto herb: a case report and review of literature. AB - The significant increase in the use of alternative medicine in general and the herbal and dietary supplement in particular represents a challenge to the health care professionals. Because of their unregulated use, physicians are encountering increasing numbers of toxicities and untoward events. We report a case of severe intraoperative haemorrhage in a patient who was taking the herb Saw Palmetto. His bleeding time which was prolonged, normalized few days after he stopped the herb. This case should increase the awareness of physicians to such possible complications and encourage them to enquire thoroughly about the use of any dietary supplement in all their patients. PMID- 11489068 TI - A case of POEMS syndrome with high concentrations of interleukin-6 in pericardial fluid. AB - The POEMS syndrome (plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy of various forms, monoclonal gammopathy, skin changes) is a rare multisystem disorder of unknown pathogenesis. Overexpression of proinflammatory cytokines has been implicated in the pathogenesis of POEMS syndrome, however, it is not known whether there is an association between abnormalities in cytokines and pericardial fluid. We present a case of POEMS syndrome with high concentrations of interleukin-6 (IL-6) in pericardial fluid. In our patient, pericarditis developed into cardiac tamponade, and the concentration of IL-6 in pericardial fluid was remarkably elevated compared with that in serum (1760 vs. 6.57 pg mL(-1)). We suggest that IL-6 is associated with the progression or maintenance of pericarditis as a result of POEMS syndrome. PMID- 11489069 TI - A case of toxic shock-like syndrome presenting with serious hypoproteinaemia because of a protein-losing gastroenteropathy. AB - A 37-year-old man was admitted to our hospital because of toxic shock-like syndrome (TSLS) induced by Streptococcus pyogenes. After the pathogenic bacteria had been eradicated, serious diarrhoea appeared and a protein-losing gastroenteropathy developed. An immunohistochemical study of the biopsy specimens of both small and large intestines revealed the infiltration of T-lymphocytes, predominantly CD8+ cells, into the lamina propria of affected mucosa, villus atrophy and crypt hyperplasia. Considering these histological findings, some immunological mechanism which lead the activation of cytotoxic T-lymphocytes may play an important role in the pathogenesis of this rare intestinal manifestation of TSLS. PMID- 11489070 TI - Secondary prevention of cardiovascular disease and diabetes: still suboptimal in patients with myocardial infarction. PMID- 11489072 TI - Electron energy-loss near-edge structure -- a tool for the investigation of electronic structure on the nanometre scale. AB - Electron energy-loss near-edge structure (ELNES) is a technique that can be used to measure the electronic structure (i.e. bonding) in materials with subnanometre spatial resolution. This review covers the theoretical principles behind the technique, the experimental procedures necessary to acquire good ELNES spectra, including potential artefacts, and gives examples relevant to materials science. PMID- 11489073 TI - Frequency-domain fluorescence microscopy with the LED as a light source. AB - We describe a frequency-domain lifetime fluorometer based on a microscope and a modulated light-emitting diode (LED) excitation source (370/460 nm), which operates in the frequency range 120 Hz--250 MHz. We collected multifrequency phase and modulation fluorescence responses from cellular areas as small as 10- 15 microm in diameter. We also collected fluorescence lifetime data from cells stained by a lipophilic coumarin sensitized europium fluorophore, Coum-Eu, with a millisecond lifetime, and Ru(bpy)(2)phe-C(12),with microsecond lifetime. Nanosecond lifetimes from native nuclei stained with SYTO 14 and SYTO 16 probes were measured as well. We demonstrate that a simple LED excitation source can, for many applications, successfully replace complex and expensive laser systems, which have been used for cellular frequency-domain lifetime measurements. As the LEDs are very stable with low noise, it will be possible to image even smaller sample areas using brighter LEDs. With availability of modulated LEDs emitting at several wavelengths covering almost the entire visible spectrum it is easy to assemble a system for the fluorophore of choice. The ability to select an excitation source for a given fluorophore and low price make such an excitation source even more practical. PMID- 11489074 TI - Nano-optical reversible switching of organic photochromes. AB - Photochromic fulgides are UHV-deposited in ultrathin films exhibiting intrinsic nanostructures. The two isomeric states (C and E) are detected in absorption via scanning near-field optical microscopy (SNOM). The optical image contrast measured in SNOM corresponds to the nanostructure topography observed in atomic force microscopy. Nano-optical reversible switching of individual nanostructures is demonstrated and possible applications to rewritable optical recording on the nanoscale are discussed. PMID- 11489076 TI - Imaging the native structure of the chaperone protein GroEL without fixation using atomic force microscopy. AB - Most sample preparation methods for scanning probe or electron microscopy require that biomolecules, such as proteins, be fixed. Fixation destroys the molecular functionality and can possibly affect the true molecular structure. Here we report sample preparation conditions that allow the imaging of an unfixed protein, GroEL, under in-vivo conditions, by atomic force microscopy. Under these conditions, the protein should maintain its native structure and biological activity. The typical toroidal shape with pore of the GroEL complex was easily visible in the images. Images of a single complex show dimensions that agree well with crystallographic data. Under in-vivo conditions, it should be possible to study the biological activity and function of proteins. PMID- 11489077 TI - Real-time confocal imaging, during active air abrasion -- substrate cutting. AB - Air abrasion cutting, using particulates accelerated in a controlled compressed gas stream, is currently being re-evaluated as a precision tissue removal technique for dental cavity preparation. The minimal vibrations and heat generated during cutting commend the technique for use in the shaping of fragile or brittle materials that are vulnerable to vibrations and thermal stresses. Traditional air abrasion studies have relied solely upon post-procedure imaging, and cutting process details have been inferred from the nature of the residual surface. In this paper, however, a real-time confocal microscopic imaging method is described, which for the first time has allowed prior target structure characterization with subsequent imaging of cutting interactions and substrate failure patterns. Using internally focusing long working distance Hill objective lenses, focusing deep to a protective microscope slide and adhesive interfaces, unhindered remote image sampling within the bulk of specimens such as tooth tissue, acrylic and brittle ceramics was possible. Moreover, areas of active cutting and inactive regions were identified within air abraded cavities during their creation. The characteristics of the finished cut surfaces were demonstrated and confirmed the findings of previous SEM studies. The method allowed direct control over all the known variables influencing cutting with particulate streams. PMID- 11489078 TI - 3D light scanning macrography. AB - The technique of 3D light scanning macrography permits the non-invasive surface scanning of small specimens at magnifications up to 200x. Obviating both the problem of limited depth of field inherent to conventional close-up macrophotography and the metallic coating required by scanning electron microscopy, 3D light scanning macrography provides three-dimensional digital images of intact specimens without the loss of colour, texture and transparency information. This newly developed technique offers a versatile, portable and cost efficient method for the non-invasive digital and photographic documentation of small objects. Computer controlled device operation and digital image acquisition facilitate fast and accurate quantitative morphometric investigations, and the technique offers a broad field of research and educational applications in biological, medical and materials sciences. PMID- 11489079 TI - Micromanipulation of chloroplasts using optical tweezers. AB - This paper describes experiments using optical tweezers to probe chloroplast arrangement, shape and consistency in cells of living leaf tissue and in suspension. Dual optical tweezers provided two-point contact on a single chloroplast or two-point contact on two adhered chloroplasts for manipulation in suspension. Alternatively, a microstirrer consisting of a birefringent particle trapped in an elliptically polarized laser trap was used to induce motion and tumbling of a selected chloroplast suspended in a solution. We demonstrate that displacement of chloroplasts inside the cell is extremely difficult, presumably due to chloroplast adhesion to the cytoskeleton and connections between organelles. The study also confirms that the chloroplasts are very thin and extremely cup-shaped with a concave inner surface and a convex outer surface. PMID- 11489080 TI - An improved method for preparing thick sections for immuno/histochemistry and confocal microscopy and its use to identify rare events. AB - Detection of rare events within solid tissues by immunocytochemistry is aided by imaging thick sections. Sections of 40--100 microm thickness of paraformaldehyde fixed solid tissue can be prepared by use of a vibrating microtome and when immunolabelled these sections can be imaged in a confocal microscope. This approach provides excellent preservation of the structure of the sample and imposes minimal antigenic damage. In studies of the invasion of the bovine intestinal epithelium by Salmonella, this method has allowed detection of individual invading bacteria within large samples. The thick vibrating microtome sections were also used for the detection of rare apoptotic cell nuclei identified by TUNEL staining. PMID- 11489081 TI - Rapid contrasting of ultrathin sections using microwave irradiation with heat dissipation. AB - The use of microwave irradiation (MWI) to accelerate fixation, dehydration and contrasting (staining) for electron microscopy has been applied to the development of rapid methods to process biological samples in electron microscopy. A simple explanation is that the reduced time in those procedures is due to heating. In this paper we propose a contrasting method for thin sections that avoids the thermal effects of MWI. Grids with thin sections of mouse kidney, the dinoflagellate Alexandrium monilatum, spermatophores of the fly Archicepsis diversiformis, the bacteria Acinetobacter calcoaceticum and Enterobacter cloacae were placed into Beem capsules and stained with uranyl acetate and lead citrate, while immersed in an ice-water bath, and irradiated for periods ranging from 30 s to 2 min. After each contrasting procedure, the Beem capsule was filled with distilled water to wash the grids under MWI with the same irradiation time as used to contrast. Good results were obtained on irradiating for 1 min and the temperature of the Beem capsule was maintained around 5 degrees C. PMID- 11489082 TI - Corticosteroid actions in the hippocampus. AB - Corticosteroid hormones can enter the brain and bind to two intracellular receptor types that regulate transcription of responsive genes: (i) the high affinity mineralocorticoid receptors and (ii) the glucocorticoid receptors with approximately 10-fold lower affinity. Although most cells in the brain predominantly express glucocorticoid receptors, principal cells in limbic structures such as the hippocampus often contain glucocorticoid as well as mineralocorticoid receptors. Recent electrophysiological studies have examined the consequences of transcriptional regulation via the two receptor types for information transfer in the hippocampus. It was found that, under resting conditions, corticosteroids do not markedly alter electrical activity. However, if neurones are shifted towards more depolarized or hyperpolarized potentials due to the action of neurotransmitters, slow and adaptive effects of the corticosteroid hormones become apparent. In general, mineralocorticoid receptor occupation maintains steady electrical activity in hippocampal neurones. Brief activation of glucocorticoid receptors leads to increased influx of calcium, which normally helps to slowly reverse temporarily raised electrical activity. These slow and persistent corticosteroid actions will alter network function within the hippocampus, thus contributing to behavioural adaptation in response to stress. Modulation of hippocampal activity by corticosteroids also affects hippocampal output (e.g. to inhibitory interneurones which control hypothalamic pituitary-adrenal axis activity). The enhanced calcium influx after glucocorticoid receptor activation can become a risk factor when cells are simultaneously exposed to strong depolarizing inputs, such as those occurring during ischaemia. Similarly, chronically elevated corticosteroid levels (or lack of corticosteroids) could endanger hippocampal cell function. The latter may contribute to the precipitation of clinical symptoms in diseases associated with chronically aberrant corticosteroid levels. PMID- 11489083 TI - Changes in rat serum corticosterone after treatment with metabotropic glutamate receptor agonists or antagonists. AB - From previous work, it appears that glutamate can activate the hypothalamic pituitary-adrenocortical (HPA) axis by an interaction at either ionotopic or metabotropic (G-protein coupled) receptors. For example, (1S,3R)-1 aminocyclopentane-1,3-dicarboxylate (ACPD), a metabotropic glutamate (mGlu) receptor agonist, has been shown to increase the levels of serum corticosterone in rats. The present study was undertaken to further characterize which of the mGlu receptors are substantially involved in control of the HPA axis. The group I mGlu receptor agonists, 3,5-dihydroxyphenylglycine (DHPG), 1S,3R-ACPD, and 2 chloro-5-hydroxyphenylglycine (CHPG) but not the inactive isomer 1R,3S-ACPD were found to dose-dependently increase serum corticosterone 1 h after intracerebroventricular (i.c.v.) injection in male rats. The relative potency, DHPG (EC50 = 520 nmol) > 1S,3R-ACPD (1.4 micromol) = CHPG (2.7 micromol) >> 1R,3S ACPD (>> 3 micromol) is consistent with activation of group I (mGlu1/5) receptors. The effects of DHPG were long lasting with substantial elevations in corticosterone remaining for at least 3 h. In a similar manner, the group III mGlu receptor agonists, L-AP4 (4-phosphono-2-aminobutyric acid) and L-SOP (serine O-phosphate), were found to increase serum corticosterone levels at 1 h. In contrast, the mGlu group II selective agonists LY354740 (10 mg/kg, i.p.) and subtype-selective doses of the group II antagonist LY341495 (1 mg/kg, i.p.) did not significantly elevate serum corticosterone. Given the group I agonists results, it was surprising to find that group I selective and mGlu1 selective antagonists given alone also increased serum corticosterone. As with the agonists, the rise in serum corticosterone with LY393675 (an mGlu1/5 antagonist, EC50 = 20 nmol, i.c.v.) and LY367385 (an mGlu1 antagonist, 325 nmol, i.c.v.) were dose-dependent and consistent with their relative affinity for the group I mGlu receptors. The selective mGlu5 antagonist MPEP [2-methyl-6 (phenylethylnyl)pyridine] increased serum corticosterone but only at high doses (> 30 mg/kg, i.p.). A model involving the high glutamatergic tone on GABAergic interneurons in the paraventricular nucleus of the hypothalamus is discussed as a possible explanation for these results. PMID- 11489084 TI - Role of extra-ovarian oestrogens in the regulation of gonadotropin releasing hormone mRNA expression in the rat brain. AB - To further understand the role of oestrogens in the regulation of gonadotropin releasing hormone (GnRH) mRNA expression in the female rat brain, the effect of EM-652.HCl, a pure anti-oestrogen, was studied in intact and ovariectomized rats as well as in rats chronically treated with a GnRH agonist D-trp6, des-Gly-NH210 GnRH ethylamide (GnRH-A), a treatment which blocks ovarian steroidogenesis. Quantitative in situ hybridization was used to measure GnRH mRNA at the cellular level in the preoptic-anterior hypothalamic area. It was found that, 49 weeks after ovariectomy (OVX), the number of silver grains per cell corresponding to GnRH mRNA was increased by 34%. Long-term administration (49 weeks) of EM-652.HCl to OVX rats resulted in a further increase (11% over the levels measured in OVX rats) in the hybridization signal. By contrast, in intact female rats, treated during 52 weeks with EM-652.HCl, a 49% increase in the GnRH hybridization signal was detected. In rats treated with GnRH-A during the same period, a 20% decrease in GnRH mRNA was observed. When EM-652.HCl was administered concomitantly with GnRH-A, a further 63% increase over the mRNA levels recorded in GnRH-A treated rats was found. Thus, long-term treatment with the anti-oestrogen EM-652.HCl can upregulate GnRH mRNA expression in intact female rats, OVX rats and female rats chronically treated with a GnRH-A. It is suggested that the pure anti-oestrogen EM-652.HCl can exert an influence on the oestrogen feedback mechanism involved in the regulation of GnRH neuronal activity by neutralizing the action of locally produced or low circulating levels of oestrogens remaining after OVX or GnRH-A treatment. PMID- 11489085 TI - Immunohistochemical localization of calbindin D-28k in the migratory pathway from the rat olfactory placode. AB - The spatiotemporal localization of calbindin D-28k (Calb), a calcium-binding protein, was examined immunohistochemically in the developing rat olfactory system with special reference to cell migration from the olfactory placode. Calb immunoreactivity was first detected at embryonic day 12 (E12) in a few cells just outside the olfactory epithelium, and at E13, Calb-immunoreactive cells were found scattered in the laminin-rich mesenchyme. By E14, Calb-immunoreactive cells had increased in number and were seen along the entire migratory route between the vomeronasal organ, a derivative of the medial olfactory pit, and the ventromedial surface of the telencephalic vesicle. Calb neurones were not seen in the olfactory epithelium, a derivative of the lateral olfactory pit. Although the distribution pattern of Calb-immunoreactive cells was similar to that of luteinizing hormone releasing hormone (LHRH)-producing neurones, which are known to originate in the vomeronasal organ and migrate into the forebrain, Calb and LHRH immunoreactivities were contained in separate neuronal populations. Calb immunoreactive cells were localized along the vomeronasal nerves, identified by labelling the vomeronasal organ with the lipophilic dye, DiI, and strongly immunoreactive for neural cell adhesion molecule (NCAM). These data strongly suggest that, in addition to LHRH neurones, the rat vomeronasal organ generates Calb-immunoreactive neurones which migrate along the vomeronasal nerves to enter the forebrain. The final fate and functional importance of these cells remains to be determined. PMID- 11489086 TI - Nitric oxide directly activates GABA(A) receptor function through a cGMP/protein kinase-independent pathway in frog pituitary melanotrophs. AB - The direct effects of nitric oxide (NO) donors and sulfhydryl-modifying agents on the GABA(A) receptor function were examined by perforated patch, whole-cell and single channel recordings in cultured frog melanotrophs. In amphotericin B perforated cells incubated with the soluble guanylyl cyclase inhibitors LY 83583 and ODQ (10-4 M each), the NO donor sodium nitroprusside (SNP) (10(-3) M) reversibly increased the current evoked by GABA (5 x 10(-6) M). In the whole-cell configuration, internal application of the oxidizing agent H2O2 (0.05%) potentiated the GABA-evoked current while the reducing agent 2-mercaptoethanol (5 x 10(-3) M) slightly decreased the current amplitude. In inside-out patches, GABA (2 x 10(-7) M) triggered single channel bursts of openings. Incubation with the NO donors SNP or DEA/NO (10(-4) M each) enhanced the open probability of the GABA(A) receptor channel but did not modify the chloride reversal potential and did not affect the conductance states. The oxidizing agents H2O2 (0.05%) or DTNB (10-4 M) mimicked the stimulatory effect of the NO donors on the open probability while the reducing compounds 2-mercaptoethanol (5 x 10(-3) M) or DTT (10(-4) M) markedly attenuated the channel activity. Potentiation of the GABA-induced single channel activity by SNP or H2O2 was blocked by 2-mercaptoethanol. Similarly, the potentiating effect produced by DEA/NO or DTNB on the open probability was reversed by DTT. In outside-out patches, incubation with SNP also significantly enhanced the open probability of single channels activated by GABA (10(-6) M). These data indicate that, in frog pituitary melanotrophs, NO potentiates the GABA evoked current independently of the cGMP/protein kinase pathway. The effect of NO can be accounted for by S-nitrosylation/oxidation of thiol groups either directly on the GABA(A) receptor subunits or on a regulatory protein tightly associated with the GABA(A) receptor. PMID- 11489087 TI - Large reduction in the number of galanin-immunoreactive cells in pancreatic islets of diabetic rats. AB - Although galanin has been shown to be present in pancreatic islet cells, there is no literature available on the pattern of distribution and the effect of galanin in the pancreas of diabetic animals or human models. The aim of this study was to examine whether galanin immunoreactivity changes after the onset of diabetes mellitus in the rat model. The present study used immunohistochemical techniques to examine the pattern of distribution of galanin-like immunoreactive cells in the pancreas of rats with streptozotocin-induced diabetes. The effect of galanin on insulin secretion from intact rat pancreatic tissue fragments was also investigated using a radioimmunoassay technique. Numerous galanin-like immunoreactive cells were observed in both the peripheral and central regions of the islet of Langerhans of normal rat pancreas. By contrast, the islets of diabetic rat pancreas contained significantly (P < 0.0001) fewer galanin-like immunoreactive cells than nondiabetic rats. Galanin was colocalized with insulin in the islets of normal and diabetic rats. Galanin had an inhibitory effect on insulin secretion from the isolated pancreatic tissue fragments of normal and diabetic rats at all concentrations (10(-12) to 10(-6) M) employed. Galanin at 10(-9) M caused a significant (P < 0.02) decrease in insulin secretion from normal rat pancreatic tissue fragments compared to basal. These observations indicate that galanin may play a significant role in the regulation of insulin secretion. PMID- 11489089 TI - Absence of androgen receptor in the growth hormone releasing hormone-containing neurones in the rat mediobasal hypothalamus. AB - Growth hormone (GH) secretory patterns are influenced by gonadal steroids, at least in part, through modulation of hypothalamic somatostatin and GH releasing hormone (GHRH) secretion. In the adult male rat, testosterone appears to stimulate somatostatin gene expression by acting directly on androgen receptors in somatostatin neurones. The mechanism by which gonadal status influences hypothalamic GHRH gene expression is less clear. Gonadectomy reduces GHRH mRNA expression in rats, and this reduction can be prevented by the administration of testosterone or partly by a nonaromatizable androgen. While these observations suggest that androgen receptors mediate the actions of gonadal steroids on GHRH gene expression, they do not provide any information about the location of the androgen receptors involved in this process. To determine whether GHRH neurones themselves express androgen receptors, we double immunolabelled hypothalamic sections from colchicine-pretreated male rats. Although there was an overlap in the anatomical distribution of GHRH and androgen receptor-containing cell bodies, none of the nearly 900 GHRH immunolabelled cells we examined in each mediobasal hypothalamus appeared to contain androgen receptors. These results suggest that GHRH-expressing neurones are not direct targets for androgens and therefore the effects of testosterone on GHRH gene expression must be produced indirectly by some other neural or endocrine intermediary process. PMID- 11489088 TI - Effect of repeated lipopolysaccharide administration on tissue cytokine expression and hypothalamic-pituitary-adrenal axis activity in rats. AB - The effects of chronic immune challenge on cytokine expression and hypothalamic pituitary-adrenal axis (HPA) axis responses to stress were studied in Wistar rats after administration of increasing doses of lipopolysaccharide (LPS). Repeated LPS (R-LPS) decreased body weight and increased adrenal weight and pituitary pro opiomelanocortin mRNA levels. LPS injection increased plasma adrenocorticotropic hormone (ACTH) and corticosterone but the effect was attenuated in R-LPS. Plasma corticosterone but not ACTH responses to restraint were also reduced in R-LPS. Basal and restraint-stimulated corticotropin releasing hormone (CRH) mRNA levels were lower in R-LPS, but responses to a new LPS injection were similar to controls. In contrast, type 1 CRH receptor (CRH-R1) mRNA responses to both LPS and restraint were blunted in R-LPS. Vasopressin mRNA levels in parvocellular neurones were higher in R-LPS, and increased further after restraint but not after a new LPS injection. Glucocorticoid receptor (GR) levels in the paraventricular nucleus (PVN) increased after a single LPS or R-LPS (24 h after the last injection) but declined after a new injection in R-LPS. Interleukin (IL) 1beta and IL-6 mRNAs increased in the pituitary, spleen and circumventricular organs after single or R-LPS, suggesting that cytokines may contribute to the activation of the HPA axis though pathways from the circumventricular organs as well as paracrine effects in the pituitary. The data show that (i) adaptation of the HPA axis during repeated LPS injection involves increases in vasopressin : CRH expression ratios in parvocellular neurones; (ii) that hypothalamic CRH and vasopressin responses to acute stimulation are independent of CRH-R1 expression in the PVN; and (iii) there is a dissociation between pituitary and adrenal responses to acute stress suggesting a decrease of adrenal sensitivity to ACTH. PMID- 11489090 TI - Alterations in hypothalamic insulin-like growth factor-I and its associations with gonadotropin releasing hormone neurones during reproductive development and ageing. AB - Insulin-like growth factor-I (IGF-I) is thought to play a role in the onset of reproductive ability at puberty and the control of reproductive function throughout adult life. It is believed that these effects are mediated at least in part by the activation of gonadotropin releasing hormone (GnRH) neurones by IGF I, but the interactions of IGF-I with GnRH neurones in vivo are largely unknown. We first examined the anatomical relationship between GnRH and IGF-I cells in neuroendocrine regions. Using double-label immunocytochemistry, we observed that in the preoptic area-anterior hypothalamus (POA-AH), the site of GnRH perikarya, the majority (78%) of GnRH cell bodies expressed IGF-I immunoreactivity. IGF-I immunoreactivity was also high in the median eminence, the site of GnRH release, and GnRH neuroterminals were seen to interweave among IGF-I-immunopositive cells. Due to this substantial overlap of GnRH and IGF-I immunoreactive elements, we then tested the hypothesis that changes in IGF-I may regulate the GnRH system. Animals were examined at the two important reproductive life transitions: puberty and reproductive senescence. IGF-I mRNA levels were measured in POA-AH and medial basal hypothalamus-median eminence (MBH-ME) and effects of IGF-I treatment on GnRH mRNA levels were quantified by RNase protection assay. Although IGF-I treatment did not alter GnRH gene expression, there were significant alterations in hypothalamic IGF-I gene expression at both puberty and reproductive senescence. During puberty, IGF-I mRNA levels in the MBH-ME of rats increased from the juvenile stage (P25) to the day of vaginal opening (P35), and from the day of vaginal opening to young adulthood (P45) in the POA-AH. During reproductive ageing, IGF-I mRNA levels were significantly lower in middle-aged than young rats, particularly in the MBH-ME. At all ages, IGF-I expression was greater in the MBH-ME than in the POA-AH. These experiments demonstrate that: (i) the majority of adult GnRH neurones are immunopositive for the IGF-I protein; (ii) hypothalamic IGF-I levels increase at the onset of reproductive function and decrease at reproductive senescence in a regionally specific manner; and (iii) despite the presence of IGF-I in GnRH perikarya, IGF-I does not affect GnRH gene expression, suggesting that IGF-I may act at the level of GnRH release rather than gene expression. PMID- 11489092 TI - More medical students for England. PMID- 11489093 TI - Why not teach where the patients are? PMID- 11489094 TI - Learning about ethics: the cardinal rule of the clinical experience. PMID- 11489095 TI - Women and medicine--a new paradigm. PMID- 11489096 TI - The validity of long cases. PMID- 11489097 TI - Community-based teaching: killing the goose that laid the golden egg? PMID- 11489098 TI - Redefining medical students' disease to reduce morbidity. AB - OBJECTIVES: To gain a clearer conceptual understanding of medical students' disease and its impact on students, by separating the process of thinking that one may have a particular illness under study from the emotional distress that may accompany these thoughts. METHOD: In a questionnaire survey, the responses of 92 first-year and 85 third-year medical students were compared with those of 82 law students, with regard to medical students' disease perception, medical students' disease distress, hypochondriacal beliefs, concerns about health, the value placed on health, and recent visits to doctors in the past 12 months. SETTING: The University of Auckland, New Zealand. RESULTS: Both groups of medical students scored higher on medical students' disease perception than law students. First-year students scored higher on medical students' disease distress and hypochondriacal concerns than both law and third-year medical students. While medical students place a higher value on health, there were no differences with regard to health visits in the past year. CONCLUSIONS: The results support the separation of medical students' disease into perceptual and emotional components. This conceptualization of medical students' disease as a normal process rather than a form of hypochondriasis may be used to brief medical students when they enter medical school, in order to reduce the distress associated with the condition. Medical students' disease can also be used as a personally relevant example in teaching about how patients make sense of symptoms. PMID- 11489099 TI - Does observation add to the validity of the long case? AB - BACKGROUND: A London medical school final MBBS examination for 155 candidates. OBJECTIVE: To investigate whether observing the student-patient interaction in a history taking (HT) long case adds incremental information to the traditional presentation component. DESIGN: A prospective study of a HT long case which included both examiner observation of the student-patient interview (Part 1) and traditional presentation to different examiners (Part 2). Checklist and global ratings of both parts were compared. Examiners were paired to estimate inter rater reliability. The students also took a 20 station Objective Structured Clinical Examination (OSCE). OUTCOME MEASURES: Correlation of (I) examiner ratings for observation and presentation of the HT long case (II) examiner pair ratings and (III) stepwise regression analysis of scores for the HT long case with OSCE scores. RESULTS: Seventy-five (48.4%) candidates had two examiner pairs marking their case history. Observation and presentation scores correlated poorly (checklist 0.38 and global 0.33). Checklist and global scores for each part correlated at higher levels (observation 0.64 and presentation 0.61). Inter-rater reliability correlations were higher for observation (checklist 0.72 and global 0.71) than for presentation (checklist 0.38 and global 0.60). When HT long case scores were correlated with OSCE scores, using stepwise regression, global presentation scores showed the highest correlation with the OSCE score (0.36) and the global observation score contributed a further 12% to the correlation (0.50). CONCLUSION: Observation of history taking in a long case appears to measure a useful and distinct component of clinical competence over and above the contribution made by the presentation. PMID- 11489100 TI - Doctors' professional values: results from a cohort study of United Kingdom medical graduates. AB - OBJECTIVES: To examine young doctors' views on a number of professional issues including professional regulation, multidisciplinary teamwork, priority setting, clinical autonomy and private practice. METHOD: Postal survey of 545 doctors who graduated from United Kingdom medical schools in 1995. RESULTS: Questionnaires were returned by 95% of the cohort (515/545). On issues of professional regulation, teamwork and clinical autonomy, the majority of doctors held views consistent with current General Medical Council guidance. The majority supported the right of doctors working in the NHS to engage in private practice. Most respondents thought that public expectations of doctors, medicine and the NHS were too high, and that some form of rationing was inevitable. On many issues there was considerable variation in attitudes on the basis of sex and intended branch of medicine. CONCLUSIONS: The results highlight the heterogeneity of the profession and the influence of specialty and gender on professional values. Doctors' attitudes had also been shaped by broader social changes, especially debates surrounding regulation of the profession, rising public expectations and the need for rationing of NHS care. PMID- 11489101 TI - The doctor-patient relationship: from undergraduate assumptions to pre registration reality. AB - OBJECTIVES: To describe the ways in which the doctor-patient relationship experienced by newly qualified pre-registration house officers (PRHOs) differed from their undergraduate expectations. DESIGN: Qualitative study in which in depth semistructured interviews were carried out with each PRHO within 4-6 weeks of the start of their first job. SETTING: Three teaching hospitals, three district general hospitals and four general practices in south-east England. PARTICIPANTS: 24 newly qualified PRHOs. RESULTS: A number of differences were identified by PRHOs. These were caused in part by the impact of factors such as the shortage of time, which could lead to emotional 'blunting'. Some PRHOs were changing their ideas about what constitutes a 'good' doctor, and were redefining the meaning of a 'professional' relationship. The relationships of PRHOs with patients were also affected by the attitudes of their senior colleagues. For example, where PRHOs tried to maintain a patient-centred relationship, they could be identified by colleagues as working too slowly. PRHOs working in general practice were able to utilize and improve their communication skills with patients, but found it difficult to transfer these skills back into the hospital setting. CONCLUSIONS: Despite receiving substantial undergraduate education on how best to communicate with patients, a variety of factors conspired to prevent hospital-based PRHOs from utilizing this information. Building on these findings, a number of recommendations are made to help improve practice. PMID- 11489102 TI - Impact of patient feedback on the interpersonal skills of general practice registrars: results of a longitudinal study. AB - CONTEXT: A general practice vocational training program. OBJECTIVES: To examine the impacts and implications of different models of systematic patient feedback on the development of general practice (GP) registrars' interpersonal skills as they progressed through a GP vocational training program. DESIGN: A longitudinal study in which GP registrars were randomly assigned to three models of patient feedback: a control group and two intervention groups. The major source of data gathering was through the Doctors' Interpersonal Skills Questionnaire (DISQ) which was administered to patients immediately after their consultation. SUBJECTS: 210 GP registrars, 104 GP supervisors and 28 156 patients. RESULTS: Multivariate analysis techniques (including repeated-measures analysis) tested the effectiveness of the interventions. Findings showed that systematic patient feedback at regular intervals throughout GP training resulted in sustained levels of interpersonal skills. The most significant gains in interpersonal skills for both intervention groups occurred in the earlier stages of general practice training. Most registrars found the experience of patient feedback useful for gaining a better understanding of their interpersonal skills and for identifying areas in which they needed to improve. GP supervisors valued the opportunity to receive patient feedback themselves and found the activity a useful adjunct to their preceptor role. CONCLUSIONS: Patients, by providing feedback on doctors' interpersonal skills, have been able to contribute to improving the quality of the patient-doctor interaction. GP registrars and their supervisors value highly the role of patient feedback in interpersonal skill development. PMID- 11489103 TI - Validation of the doctor-patient communication component of the Educational Commission for Foreign Medical Graduates Clinical Skills Assessment. AB - BACKGROUND: The pivotal role of doctor-patient communication in effective health care delivery led the Educational Commission for Foreign Medical Graduates (ECFMG) to incorporate the assessment of interpersonal skills and spoken English proficiency into its Clinical Skills Assessment (CSA). Furthermore, it was decided that to pass the CSA, a candidate would need to meet or surpass defined performance standards for doctor-patient communication as a discrete component. This requirement, among others, is designed to ensure the readiness of graduates of foreign medical schools (FMGs) to enter postgraduate medical education programmes in the United States. OBJECTIVE: The primary focus of this study was to determine the extent to which performance in a simulated testing environment is related to performance in the clinical setting. METHOD: Nurses were trained to rate the communication skills of residents from the patient's perspective. A total of 43 first-year residents were evaluated. The survey ratings (n=225) were compared with the residents' CSA communication scores. RESULTS: Corrected correlations between CSA ratings and those obtained from nurses ranged from 0.61 to 0.73. CONCLUSION: This study provides evidence for the validity of the communication ratings provided by standardized patients. The reasonably strong associations between ratings obtained during testing and those obtained through observation of 'real' patient interactions suggest that external observers can provide accurate evaluations of doctor-patient communication. PMID- 11489104 TI - Assessment of English proficiency in international medical graduates by physician examiners and standardized patients. AB - CONTEXT: Since 1986, the Ontario Ministry of Health has provided a medical licensure preparation programme for international medical graduates. Because of the diversity in candidates' oral English proficiency, this competency has been viewed as a particularly important selection criterion. OBJECTIVES: To assess and compare the quality of ratings of oral English proficiency of international medical graduates provided by physician examiners and by standardized patients (SPs). PARTICIPANTS AND MATERIALS: The study samples consisted of 73 candidates for the Ontario International Medical Graduate (IMG) Program, and physician examiners and SPs in five 10-minute encounter objective structured clinical examination (OSCE) stations. Materials used were a seven-item speaking performance rating instrument prepared for the Ontario IMG Program. METHODS: Rating sheets were scanned and the results analysed using SPSS 9.0 for Windows. RESULTS: Correlations between the physician and SP ratings on the seven items ranged from 0.52 to 0.70. The SPs provided more lenient ratings. Mean alpha reliability for the physicians' ratings on the seven items was 0.59, and for the SPs' 0.64. There was poor agreement between the two sets of raters in identifying problematic candidates. CONCLUSIONS: Notwithstanding the sizable correlations between the ratings provided by the two rater groups, the results demonstrated that there was little agreement between the two groups in identifying the potentially problematic candidates. The physicians were less prone than the SPs to rate candidates as problematic. SPs may be better placed than the physician examiners to directly assess IMG candidates' oral English proficiency. PMID- 11489105 TI - Evaluating the spoken English proficiency of graduates of foreign medical schools. AB - PURPOSE: The purpose of this study was to gather additional evidence for the validity and reliability of spoken English proficiency ratings provided by trained standardized patients (SPs) in high-stakes clinical skills examination. METHOD: Over 2500 candidates who took the Educational Commission for Foreign Medical Graduates' (ECFMG) Clinical Skills Assessment (CSA) were studied. The CSA consists of 10 or 11 timed clinical encounters. Standardized patients evaluate spoken English proficiency and interpersonal skills in every encounter. Generalizability theory was used to estimate the consistency of spoken English ratings. Validity coefficients were calculated by correlating summary English ratings with CSA scores and other external criterion measures. Mean spoken English ratings were also compared by various candidate background variables. RESULTS: The reliability of the spoken English ratings, based on 10 independent evaluations, was high. The magnitudes of the associated variance components indicated that the evaluation of a candidate's spoken English proficiency is unlikely to be affected by the choice of cases or SPs used in a given assessment. Proficiency in spoken English was related to native language (English versus other) and scores from the Test of English as a Foreign Language (TOEFL). DISCUSSION: The pattern of the relationships, both within assessment components and with external criterion measures, suggests that valid measures of spoken English proficiency are obtained. This result, combined with the high reproducibility of the ratings over encounters and SPs, supports the use of trained SPs to measure spoken English skills in a simulated medical environment. PMID- 11489106 TI - The clinical and educational experiences of pre-registration house officers in general practice. AB - AIMS: To describe aspects of the clinical experience and educational supervision gained by pre-registration house officers (PRHOs) in general practice, and to relate these to the current General Medical Council (GMC) aims for general clinical training in general practice. DESIGN: Qualitative evaluation, part of which involved semistructured interviews with 12 PRHOs who were experiencing a general practice rotation. Interviews were conducted at the beginning and the end of the pre-registration year, and following return to hospital work after completion of the general practice placement. SETTINGS: Three teaching hospitals, two district general hospitals and six general practices in south-east England. PARTICIPANTS: 12 PRHOs who were involved in rotations incorporating a general practice placement. RESULTS: To varying degrees, the GMC aims for training in general practice were met for all the participants. All PRHOs recognized the value of the clinical experience and educational supervision they received in general practice. They particularly valued aspects such as having an individual training programme based on their own needs, and the interlinking of theory and practice, which aided learning. Most felt that having responsibility for their own patients acted as an important incentive for learning, and in general, PRHOs appreciated having the time to learn which general practice allowed. CONCLUSIONS: For the majority of PRHOs, the time spent in general practice was seen as a positive clinical and educational experience. In a variety of ways, the general practice placement encouraged PRHOs to develop the self-directed learning skills seen as essential to the lifelong learning advocated by the GMC. A number of recommendations are made to help improve the integration of the hospital and general practice components of these rotations. PMID- 11489107 TI - Evaluating primary care as a base for medical education: the report of the Cambridge Community-based Clinical Course. AB - BACKGROUND: Medical students receive increasing amounts of their clinical education in a primary care setting. The educational possibilities of such attachments are still being explored. AIM: To report the evaluation of a small, radical innovation to provide students with a long-term community attachment which integrated with hospital-based education. METHOD: Between 1993 and 1998, 13 medical students completed 15-month attachments with a single general practice in England. The course offered them experience in the major clinical specialties throughout this period. Students were exposed to specialist as well as generalist education in the context of patients with whom they could establish a continuing relationship. The innovation was evaluated by its feasibility, by students' examination results, by analysis of clinical experience, through formal student feedback and by cost. RESULTS: The course was practicable in a particular setting with academic leadership. The students all passed their exams. They had wide, appropriate clinical experience even though the attachment was to a single practice. When they returned to the hospital environment, students did not feel themselves at a disadvantage compared with traditional students. The costs of the course are controversial: placement costs were higher than in the hospital, but those for facilities were lower. CONCLUSION: It is possible to run a course like this successfully. It remains the most radical attempt to share clinical education in the UK between primary and secondary/tertiary care. Further research is required into providing long-term clinical attachments in NHS primary care settings. PMID- 11489108 TI - A survey of primary care physicians' perceptions and needs regarding the precepting of medical students in their offices. AB - OBJECTIVE: To assess the interest, perceptions, and needs of primary care physicians with regard to office-based precepting of medical students. DESIGN: Random survey. SETTING: The New England region of the United States (Maine, New Hampshire, Vermont, Massachusetts, Rhode Island, Connecticut). PARTICIPANTS: Family physicians, general internists and paediatricians. MAIN OUTCOME MEASURES: These included: (1) practice and preceptor demographics; (2) Likert scale agreement or disagreement with various positive and negative perceptions about precepting students; rating (from unimportant to necessary) of potential benefits from medical schools associated with the support of precepting in the office setting, and (3) comparisons among physician groups to determine differences in perceptions and needs. RESULTS: There is a high level of interest in precepting regardless of primary care specialty, practice structure, payment mechanism, or precepting experience. Negative impacts included decreased productivity and increased length of the day by a median of 60 minutes. Positive impacts included keeping one's own knowledge up-to-date and enhanced enjoyment of practice. Benefits such as academic appointments, continuing medical education credits, faculty development, computer linkages for clinical information and medical library access are rated highly. Monetary payment, whether as a modest honorarium or as compensation for lost time/income, was felt to be important by half of our sample. CONCLUSION: A significant interest in precepting medical students on a regular basis is expressed by primary care physicians in the ambulatory medicine setting. The results of this survey can be used by medical schools to address negative perceptions and to develop appropriate benefits packages to recruit and retain these primary care preceptors. PMID- 11489109 TI - Perspective on South America: the Latin American contribution to the world movement in medical education. AB - The invigoration of Latin American medical education during the past decade has been remarkable. The new initiatives which have taken place and the innovative programmes which have been enacted are analysed with reference to the seminal participation in international ventures. The analysis demonstrates that, while South American regional development was undeniably and profoundly influenced by the world movement in medical education, there has also been a reciprocal influence. South America has contributed notably to global action. The extent of the contribution by South America to the world movement, and the benefits gained in turn, make it self-evident that continuation of such bilateral exchange is crucial, and is to be energetically promoted. PMID- 11489110 TI - Resuscitating paediatric induction: an action research approach. AB - OBJECTIVES: To establish teaching objectives, methods and assessment for a paediatric induction programme. DESIGN: Action research over a 2-year period. PARTICIPANTS: 88 doctors from three hospitals. MAIN OUTCOME MEASURES: These were end-of-course evaluations, using Likert scales, and free comments; independent evaluation by the West Midlands Deanery 4 months after courses, in which participating and non-participating units were compared, and summative assessment of trainees involving multiple-choice questions, a basic life-support scenario and an illness scenario. RESULTS: 88 participants attended four 3-day courses over 2 years. Mean satisfaction ratings were very high for content, personal value, and presentation. There was a significant rise in Deanery evaluation scores, compared with non-significant change in other hospitals, and there were excellent pass rates for trainees. CONCLUSIONS: Trainees strongly approve of induction with clear objectives, active teaching, and validated assessment. PMID- 11489111 TI - Change management in primary care: design and evaluation of an internet-delivered course. AB - OBJECTIVES: To deliver and evaluate an internet course in change management for primary care professionals. DESIGN: A 12-week course delivered over the internet. Respondents were allocated into two groups: one had access to the course tutor individually, while the other could also communicate with the rest of the learner group. Learning outcomes were assessed by means of pre- and post-intervention questionnaires and by interviews with some participants. SUBJECTS: 111 primary care professionals. RESULTS: Subjects showed significant improvements on all learning outcomes. The group with access to the course tutor alone completed more units and had greater improvements in their learning outcomes. More of this group completed the course and completed a portfolio. Respondents who could communicate with other members of the group did not find this a positive experience. CONCLUSION: The internet can be used to deliver learning in change management to primary care professionals. Access to a discussion forum did not improve, and possibly impaired learning. More work is needed on the mentoring of internet learning. PMID- 11489112 TI - CeMENT: a multidisciplinary teaching programme. PMID- 11489113 TI - Evaluation of a 'breaking bad news' course at the Charite, Berlin. PMID- 11489114 TI - Teaching international health issues to medical students. PMID- 11489115 TI - Beyond biomedicine: the challenge of teaching preventive medicine. PMID- 11489116 TI - Negotiated learning for generic teaching in palliative care within general practice teaching. PMID- 11489117 TI - Colombian physician brain drain. PMID- 11489118 TI - Evolution of a molecular switch: universal bacterial GTPases regulate ribosome function. AB - The GTPases comprise a protein superfamily of highly conserved molecular switches adapted to many diverse functions. These proteins are found in all domains of life and often perform essential roles in fundamental cellular processes. Analysis of data from genome sequencing projects demonstrates that bacteria possess a core of 11 universally conserved GTPases (elongation factor G and Tu, initiation factor 2, LepA, Era, Obg, ThdF/TrmE, Ffh, FtsY, EngA and YchF). Investigations aimed at understanding the function of GTPases indicate that a second conserved feature of these proteins is that they elicit their function through interaction with RNA and/or ribosomes. An emerging concept suggests that the 11 universal GTPases are either necessary for ribosome function or transmitting information from the ribosome to downstream targets for the purpose of generating specific cellular responses. Furthermore, it is suggested that progenitor GTPases were early regulators of RNA function and may have existed in precursors of cellular systems driven by catalytic RNA. If this is the case, then a corollary of this hypothesis is that GTPases that do not bind RNA arose at a later time from an RNA-binding progenitor that lost the capability to bind RNA. PMID- 11489119 TI - The nsdD gene encodes a putative GATA-type transcription factor necessary for sexual development of Aspergillus nidulans. AB - The ability to reproduce both sexually and asexually is one of the characteristics of the homothalic ascomycete Aspergillus nidulans. Unlike the other Aspergillus species, A. nidulans undergoes sexual development that seems to be regulated by internal and external stimuli. To begin to understand the sexual reproduction of A. nidulans we previously isolated and characterized several NSD (never in sexual development) mutants that failed to produce any sexual reproductive organs, and identified four complementation groups, nsdA, nsdB, nsdC, and nsdD. The nsdD gene has been isolated, and it is predicted to encode a GATA-type transcription factor with the type IVb zinc finger DNA-binding domain. The mRNA of the nsdD gene started to accumulate in the early phase of vegetative growth, and the level increased as sexual development proceeded. However, it decreased during asexual sporulation and no nsdD mRNA was detected in conidia. Deletion of nsdD resulted in no cleistothecia (fruiting bodies) formation, even under the conditions that preferentially promoted sexual development, indicating that nsdD is necessary for sexual development. In contrast, when the nsdD gene was over-expressed, sexual-specific organ (Hulle cell) was formed even in submerged culture, which normally completely blocked sexual development, and the number of cleistothecia was also dramatically increased on solid medium. These results lead us to propose that the nsdD gene functions in activating sexual development of A. nidulans. Multiple copies of the nsdD gene could suppress nsdB5 and veA1, indicating that either nsdD acts downstream of these genes or possibly functions in overlapping pathway(s). PMID- 11489120 TI - Evidence for a rolling-circle mechanism of phage DNA synthesis from both replicative and integrated forms of CTXphi. AB - The genes encoding cholera toxin, the principal virulence factor of Vibrio cholerae, are part of the circular single-stranded DNA genome of CTXphi. In toxigenic V. cholerae strains, the CTXphi genome is typically found in integrated arrays of tandemly arranged CTX prophages. Infected cells that lack a chromosomal integration site harbour the CTXphi genome as a plasmid (pCTX). We studied the replication of pCTX and found several indications that this plasmid replicates via a rolling-circle (RC) mechanism. The initiation and termination sites for pCTX plus-strand DNA synthesis were mapped to a 22 bp sequence that contains inverted repeats and a nonanucleotide motif found in the plus-strand origins of several RC replicons. Furthermore, similar to other RC replicons, replication of plasmids containing duplicated pCTX origins resulted in the deletion of sequences between the two origins and the formation of a single chimeric origin. Our previous work revealed that CTX prophage arrays give rise to hybrid CTX virions that contain sequences derived from two adjacent prophages. We now report that the boundaries between the sequences contributed to virions by the upstream and the downstream prophages in an array correspond to the site at which synthesis of plus-strand pCTX DNA is initiated and terminated. These data support the model that plus-strand CTXphi DNA is generated from chromosomal prophages via a novel process analogous to RC replication. PMID- 11489121 TI - Identification of a genetic determinant responsible for host specificity in Streptococcus thermophilus bacteriophages. AB - Phage-host interactions remain poorly understood in lactic acid bacteria and essentially in all Gram-positive bacteria. The aim of this study was to identify the phage genetic determinant (anti-receptor) involved in the recognition of Streptococcus thermophilus hosts. The complete genomic sequence of the lytic S. thermophilus phage DT1 was determined previously, and bioinformatic analysis indicated that orf18 might be the anti-receptor gene. The orf18 of six additional S. thermophilus phages was determined (DT2, DT4, MD1, MD2, MD4 and Q5) and compared with the orf18 of DT1. The deduced ORF18 was divided into three domains. The first domain, which contains the N-terminal part of the protein, was conserved in all seven phages. The second domain was detected in only two phages and flanked by a motif called collagen-like repeats. The second domain also contained a variable region (VR1). All seven phages had a third domain that consisted of the C-terminal section of the protein as well as another variable region (VR2). Chimeric DT1 phages were constructed by recombination; a portion of its orf18 was replaced by the corresponding section in orf18 of the phage MD4. All DT1 chimeric phages acquired the host range of phage MD4. Analysis of the orf18 in the chimeric phages revealed that host specificity in phages DT1 and MD4 resulted from VR2. This is the first report on the identification and characterization of a phage gene involved in the host recognition process of Gram positive bacteria. PMID- 11489122 TI - The hgl1 gene is required for dimorphism and teliospore formation in the fungal pathogen Ustilago maydis. AB - The fungal pathogen Ustilago maydis causes a dramatic disease in maize involving the induction of tumours and the formation of masses of black teliospores. In this fungus, mating between haploid, budding cells results in the formation of the infectious, filamentous cell type that invades host tissue. Mating and filamentous growth are governed by the mating-type loci and by cAMP signalling, perhaps in response to signals from maize. To dissect the involvement of cAMP signalling further, the constitutive filamentous phenotype of a mutant with a defect in the catalytic subunit of protein kinase A was used to isolate suppressor mutations that restore budding growth. One such mutation identified the hgl1 gene, which is shown to be required for both the switch between budding and filamentous growth and teliospore formation during infection. In addition, the hgl1 gene product may be a target of phosphorylation by protein kinase A, and transcript levels for the gene are elevated during mating. Thus, the hgl1 gene provides a connection between mating, cAMP signalling and two important aspects of virulence: filamentous growth and the formation of teliospores. PMID- 11489123 TI - MlrA, a novel regulator of curli (AgF) and extracellular matrix synthesis by Escherichia coli and Salmonella enterica serovar Typhimurium. AB - Production of curli (AgF) adhesins by Escherichia coli and Salmonella enterica serovar Typhimurium (S. typhimurium) is associated with extracellular matrix production and is optimal at low temperature during stationary phase. Curli and extracellular matrix synthesis involves a complex regulatory network that is dependent on the CsgD (AgfD) regulator. We have identified a novel regulator, termed MlrA, that is required for curli production and extracellular matrix formation. Two cosmids from a genomic library of avian pathogenic E. coli chi7122 conferred mannose-resistant haemagglutination (HA) and curli production to E. coli HB101, which is unable to produce curli owing to a defective regulatory pathway. The rpoS gene, encoding a known positive regulator of curli synthesis, and the E. coli open reading frame (ORF) of unknown function, yehV, identified on each of these cosmids, respectively, conferred curli production and HA to E. coli HB101. We have designated yehV as the mlrA gene for MerR-like regulator A because its product shares similarities with regulatory proteins of the MerR family. HA and curli production by strain chi7122 were abolished by disruption of rpoS, mlrA or csgA, the curli subunit gene. Both csgD and csgBA transcription, required for expression of curli, were inactive in an mlrA mutant grown under conditions that promote curli production. An mlrA homologue was identified in S. typhimurium. Analysis of mlrA-lac operon fusions demonstrated that mlrA was positively regulated by rpoS. mlrA mutants of wild-type S. typhimurium SL1344 or SR-11 no longer produced curli or rugose colony morphology, and exhibited enhanced aggregation and extracellular matrix formation when complemented with the mlrA gene from either S. typhimurium or E. coli present on a low-copy-number plasmid. However, inactivation of mlrA did not affect curli production and aggregative morphology in an upregulated curli producing S. typhimurium derivative containing a temperature- and RpoS-independent agfD promoter region. These results indicate that MlrA is a newly defined transcriptional regulator of csgD/agfD that acts as a positive regulator of RpoS-dependent curli and extracellular matrix production by E. coli and S. typhimurium. PMID- 11489124 TI - Molecular genetics of SaPI1--a mobile pathogenicity island in Staphylococcus aureus. AB - The Staphylococcus aureus gene for toxic shock toxin (tst) is carried by a 15 kb mobile pathogenicity island, SaPI1, that has an intimate relationship with temperate staphylococcal phage 80alpha. During phage growth, SaPI1 is excised from its unique chromosomal site, attC, replicates autonomously, interferes with phage growth, and is efficiently encapsidated into special small phage heads commensurate with its size. Upon transfer to a recipient organism, SaPI1 integrates at attC by means of a self-coded integrase. One or more phage functions are required for excision, autonomous replication and encapsidation of the element and, thus, the overall relationship between SaPI1 and 80alpha is similar to that between coliphages P4 and P2. Among other staphylococcal phages tested, only phi13 interacts with SaPI1, inducing excision but not replication or transfer of the element. PMID- 11489125 TI - Natural transformation competence in Helicobacter pylori is mediated by the basic components of a type IV secretion system. AB - Helicobacter pylori (Hp), a Gram-negative bacterial pathogen and aetiologic agent of gastroduodenal disease in humans, is naturally competent for genetic transformation. Natural competence in bacteria is usually correlated with the presence of type IV pili or type IV pilin-like proteins, which are absent in Hp. Instead, we recently identified the comB operon in Hp, carrying four genes tentatively designated as orf2, comB1, comB2 and comB3. We show here that all ComB proteins and the 37-amino-acid Orf2 peptide display significant primary sequence and structural homology/identity to the basic components of a type IV secretion apparatus. ComB1, ComB2 and ComB3, now renamed ComB8, ComB9 and ComB10, correspond to the Agrobacterium tumefaciens VirB8, VirB9 and VirB10 proteins respectively. The peptide Orf2 carries a lipoprotein motif and a second cysteine residue homologous to VirB7, and was thus designated ComB7. The putative ATPase ComB4, encoded by the open reading frame hp0017 of strain 26695, corresponds to virB4 of the A. tumefaciens type IV secretion system. A Hp comB4 transposon insertion mutant was totally defective in natural transformation. By complementation of a Hp DeltacomB deletion mutant, we demonstrate that each of the proteins from ComB8 to ComB10 is absolutely essential for the development of natural transformation competence. The putative lipoprotein ComB7 is not essential, but apparently stabilizes the apparatus and modulates the transformation efficiency. Thus, pathogenic type I Hp strains contain two functional independent type IV transport systems, one for protein translocation encoded by the cag pathogenicity island and one for uptake of DNA by natural transformation. The latter system indicates a possible novel mechanism for natural DNA transformation in bacteria. PMID- 11489126 TI - Overlapping binding sites for the virulence gene regulators AphA, AphB and cAMP CRP at the Vibrio cholerae tcpPH promoter. AB - The expression of the Vibrio cholerae virulence factors, toxin-co-regulated pilus (TCP) and cholera toxin (CT), are dependent on the ability of the LysR regulator AphB to co-operate with a second protein, AphA, to activate the expression of the membrane-bound transcription factors TcpP and TcpH. To gain insights into the mechanism by which AphA and AphB co-operate to activate the expression of tcpPH, we have purified these two proteins to near homogeneity and show that they are each capable of interacting with the classical tcpPH promoter at distinct binding sites. As shown by tcpP-lacZ promoter deletion experiments, gel shift and DNase I footprinting, AphA binds to and activates from a region of the promoter between 101 and -71 from the start of transcription. AphB binds to and activates from a partially overlapping downstream site between -78 and -43, and these functions are dependent upon a region of partial dyad symmetry that resembles the well characterized LysR-binding motif. A single basepair difference in this region of dyad symmetry has been shown previously to play a critical role in the expression of virulence genes between the two disease-causing biotypes of V. cholerae, classical and El Tor. We also show here that the tcpPH promoter is negatively influenced by the global regulator cAMP-CRP. Purified CRP binds to a near consensus sequence in the tcpPH promoter in a cAMP-dependent manner and protects from DNase I digestion a region that is completely within the region protected by AphA and AphB. These findings raise the possibility that the negative effect of cAMP-CRP on virulence gene expression is the result of its ability to influence AphA- and AphB-dependent transcriptional activation of tcpPH under various conditions. PMID- 11489127 TI - Transcription of glycolytic genes and operons in Bacillus subtilis: evidence for the presence of multiple levels of control of the gapA operon. AB - Glycolysis is one of the main pathways of carbon catabolism in Bacillus subtilis. Although the biochemical activity of glycolytic enzymes has been studied in detail, no information about the expression of glycolytic genes has so far been available in this organism. Therefore, transcriptional analysis of all glycolytic genes was performed. The genes cggR, gapA, pgk, tpi, pgm and eno, encoding the enzymes required for the interconversion of triose phosphates, are transcribed as a hexacistronic operon as demonstrated by Northern analysis. This gapA operon is repressed by the regulator CggR. The presence of sugars and amino acids synergistically results in the induction of the gapA operon. The transcriptional start site upstream of cggR was mapped by primer extension. Transcripts originating upstream of cggR are processed near the 3' end of cggR. This endonucleolytic cleavage leads to differential stability of the resulting processing products: the monocistronic cggR message is very rapidly degraded, whereas the mRNA species encoding glycolytic enzymes exhibit much higher stability. An additional internal constitutive promoter was identified upstream of pgk. Thus, gapA is the most strongly regulated gene of this operon. The pfk pyk operon encoding phosphofructokinase and pyruvate kinase is weakly induced by glucose. In contrast, the genes pgi and fbaA, coding for phosphoglucoisomerase and fructose-1,6-bisphosphate aldolase, are constitutively expressed. PMID- 11489128 TI - The Mycobacterium tuberculosis ECF sigma factor sigmaE: role in global gene expression and survival in macrophages. AB - In previously published work, we identified three Mycobacterium tuberculosis sigma (sigma) factor genes responding to heat shock (sigB, sigE and sigH). Two of them (sigB and sigE) also responded to SDS exposure. As these responses to stress suggested that the sigma factors encoded by these genes could be involved in pathogenicity, we are studying their role in physiology and virulence. In this work, we characterize a sigE mutant of M. tuberculosis H37Rv. The sigE mutant strain was more sensitive than the wild-type strain to heat shock, SDS and various oxidative stresses. It was also defective in the ability to grow inside both human and murine unactivated macrophages and was more sensitive than the wild-type strain to the killing activity of activated murine macrophages. Using microarray technology and quantitative reverse transcription-polymerase chain reaction (RT-PCR), we started to define the sigmaE regulon of M. tuberculosis and its involvement in the global regulation of the stress induced by SDS. We showed the requirement for a functional sigE gene for full expression of sigB and for its induction after SDS exposure but not after heat shock. We also identified several genes that are no longer induced when sigmaE is absent. These genes encode proteins belonging to different classes including transcriptional regulators, enzymes involved in fatty acid degradation and classical heat shock proteins. PMID- 11489129 TI - Haemophore-mediated bacterial haem transport: evidence for a common or overlapping site for haem-free and haem-loaded haemophore on its specific outer membrane receptor. AB - Bacterial extracellular haemophores also named HasA for haem acquisition system form an independent family of haemoproteins that take up haem from host haeme carriers and shuttle it to specific receptors (HasR). Haemophore receptors are required for the haemophore-dependent haem acquisition pathway and alone allow free or haemoglobin-bound haem uptake, but the synergy between the haemophore and its receptor greatly facilitates this uptake. The three-dimensional structure of the Serratia marcescens holo-haemophore (HasASM) has been determined previously and revealed that the haem iron atom is ligated by tyrosine 75 and histidine 32. The phenolate of tyrosine 75 is also tightly hydrogen bonded to the Ndelta atom of histidine 83. Alanine mutagenesis of these three HasASM residues was performed, and haem-binding constants of the wild-type protein, the three single mutant proteins, the three double mutant proteins and the triple mutant protein were compared by absorption spectrometry to probe the roles of H32, Y75 and H83 in haem binding. We show that one axial iron ligand is sufficient to ligate haem efficiently and that H83 may become an alternative iron ligand in the absence of Y75 or both H32 and Y75. All the single mutant proteins retained the ability to stimulate haemophore-dependent haem uptake in vivo. Thus, the residues H32, Y75 and H83 are not individually necessary for haem delivery to the receptor. The binding of haem-free and haem-loaded HasASM proteins to HasRSM-producing strains was studied. Both proteins bind to HasRSM with similar apparent Kd. The double mutant H32A-Y75A competitively inhibits binding to the receptor of both holo HasASM and apo-HasASM, showing that there is a unique or overlapping site on HasRSM for the apo- and holo-haemophores. Thus, we propose a new mechanism for haem uptake, in which haem is exchanged between haem-loaded haemophores and unloaded haemophores bound to the receptor without swapping of haemophores on the receptor. PMID- 11489130 TI - Characterization of integration host factor (IHF) binding upstream of the cysteine-rich protein operon (omcAB) promoter of Chlamydia trachomatis LGV serovar L2. AB - Chlamydiae are bacterial parasites that carry out a distinct developmental cycle within host cells; however, the mechanisms by which these organisms regulate stage-specific gene expression are not known. We identified a DNA element located between nucleotide (nt) -135 and -90 upstream from the transcription start point of the late stage-specific CRP operon (omcAB) of Chlamydia trachomatis, to which a protein in extracts of chlamydiae harvested at 23 h after infection binds. A recombinant protein of C. trachomatis open reading frame (ORF) CT267, which is homologous to bacterial integration host factor (IHF) and the heat-unstable nucleoid protein (HU), bound to the same element and produced the same DNase I footprint as the protein in chlamydial extracts. Recombinant ORF CT267 protein bound with high affinity to the DNA element and induced a sharp bend in a DNA fragment containing the binding site, suggesting that ORF CT267 encodes a protein with IHF-like activity, and recombinant protein had a positive effect on in vitro transcription of the CRP operon. IHF-binding activity and IHF protein were detected in extracts of C. trachomatis during the early to intermediate phases of the late stage of the developmental cycle (between 17 and 30 h after infection), but were absent in the extreme late phase of the cycle and in the infectious form of chlamydiae. The presence of an IHF binding site upstream of the CRP operon and the presence of chlamydial IHF-like protein when late stage genes are transcribed suggests that the chlamydial IHF may play a role in stage-specific gene expression. PMID- 11489131 TI - The LuxS family of bacterial autoinducers: biosynthesis of a novel quorum-sensing signal molecule. AB - Many bacteria control gene expression in response to cell population density, and this phenomenon is called quorum sensing. In Gram-negative bacteria, quorum sensing typically involves the production, release and detection of acylated homoserine lactone signalling molecules called autoinducers. Vibrio harveyi, a Gram-negative bioluminescent marine bacterium, regulates light production in response to two distinct autoinducers (AI-1 and AI-2). AI-1 is a homoserine lactone. The structure of AI-2 is not known. We have suggested previously that V. harveyi uses AI-1 for intraspecies communication and AI-2 for interspecies communication. Consistent with this idea, we have shown that many species of Gram negative and Gram-positive bacteria produce AI-2 and, in every case, production of AI-2 is dependent on the function encoded by the luxS gene. We show here that LuxS is the AI-2 synthase and that AI-2 is produced from S-adenosylmethionine in three enzymatic steps. The substrate for LuxS is S-ribosylhomocysteine, which is cleaved to form two products, one of which is homocysteine, and the other is AI 2. In this report, we also provide evidence that the biosynthetic pathway and biochemical intermediates in AI-2 biosynthesis are identical in Escherichia coli, Salmonella typhimurium, V. harveyi, Vibrio cholerae and Enterococcus faecalis. This result suggests that, unlike quorum sensing via the family of related homoserine lactone autoinducers, AI-2 is a unique, 'universal' signal that could be used by a variety of bacteria for communication among and between species. PMID- 11489132 TI - Identification of the Helicobacter pylori anti-sigma28 factor. AB - Flagellar motility is essential for colonization of the human gastric mucosa by Helicobacter pylori. The flagellar filament is composed of two subunits, FlaA and FlaB. Transcription of the genes encoding these proteins is controlled by the sigma28 and sigma54 factors of RNA polymerase respectively. The expression of flagellar genes is regulated, but no sigma28-specific effector was identified. It was also unclear whether H. pylori possessed a checkpoint for flagellar synthesis, and no gene encoding an anti-sigma28 factor, FlgM, could be identified by sequence similarity searches. To investigate the sigma28-dependent regulation, a new approach based on genomic data was used. Two-hybrid screening with the H. pylori proteins identified a protein of unknown function (HP1122) interacting with the sigma28 factor and defined the C-terminal part of HP1122 (residues 48 76) as the interaction domain. HP1122 interacts with region 4 of sigma28 and prevents its association with the beta-region of H. pylori RNA polymerase. Thus, HP1122 presented the characteristics of an anti-sigma28 factor. This was confirmed in H. pylori by RNA dot-blot hybridization and electron microscopy. The level of sigma28-dependent flaA transcription was higher in a HP1122-deficient strain and was decreased by the overproduction of HP1122. The overproduction of HP1122 also resulted in H. pylori cells with highly truncated flagella. These results demonstrate that HP1122 is the H. pylori anti-sigma28 factor, FlgM, a major regulator of flagellum assembly. Potential anti-sigma28 factors were identified in Campylobacter jejuni, Pseudomonas aeruginosa and Thermotoga maritima by sequence homology with the C-terminal region of HP1122. PMID- 11489133 TI - Genetic analysis of the signalling pathway activated by external amino acids in Saccharomyces cerevisiae. AB - The permease-like amino acid sensor Ssy1p of Saccharomyces cerevisiae is required for transcriptional induction, in response to external amino acids, of several genes encoding peptide and amino acid permeases. Among them is AGP1 encoding a low-affinity, broad-specificity amino acid permease important for the utilization of amino acids as a nitrogen source. We report here data from experiments aimed at identifying components of the signalling pathway activated by Ssy1p. Overproduction of the large amino-terminal tail of Ssy1p interferes negatively with the induction of AGP1 in wild-type cells. Furthermore, overproduction of this domain can relieve growth defects of a ssy1 null strain, indicating that the N-terminal tail of Ssy1p is an important functional element of the pathway. Consistent with a role for Ssy1p in the recognition of amino acids, a mutant form of the protein with a Thr to Ile substitution in the eighth predicted transmembrane domain is competent for the induction of AGP1 by leucine but not by other amino acids. In a screen for other mutants defective in the Ssy1p pathway, we confirmed that PTR3 and SSY5 encode additional factors essential for AGP1 expression in response to multiple amino acids. Data obtained by overproducing Ptr3p and Ssy5p in ssy1Delta, ptr3Delta and ssy5Delta mutants suggest that Ptr3p acts downstream from Ssy1p and Ssy5p downstream from Ptr3p in the transduction pathway. Furthermore, two-hybrid experiments indicated that Ptr3p interacts with Ssy5p and that Ptr3p can self-associate. Finally, the Cys-6-Zn2 transcription factor Uga35p/Dal81p required for the induction of AGP1 is also essential for the expression of two other genes under Ssy1p-Ptr3p-Ssy5p control, namely BAP2 and PTR2, suggesting that the protein is yet another component of the amino acid signalling pathway. PMID- 11489134 TI - Structure, activity and evolution of the group I thiolactone peptide quorum sensing system of Staphylococcus aureus. AB - In Staphylococcus aureus, the agr locus is responsible for controlling virulence gene expression via quorum sensing. As the blockade of quorum sensing offers a novel strategy for attenuating infection, we sought to gain novel insights into the structure, activity and turnover of the secreted staphylococcal autoinducing peptide (AIP) signal molecules. A series of analogues (including the L-alanine and D-amino acid scanned peptides) was synthesized to determine the functionally critical residues within the S. aureus group I AIP. As a consequence, we established that (i) the group I AIP is inactivated in culture supernatants by the formation of the corresponding methionyl sulphoxide; and (ii) the group I AIP lactam analogue retains the capacity to activate agr, suggesting that covalent modification of the AgrC receptor is not a necessary prerequisite for agr activation. Although each of the D-amino acid scanned AIP analogues retained activity, replacement of the endocyclic amino acid residue (aspartate) located C terminally to the central cysteine with alanine converted the group I AIP from an activator to a potent inhibitor. The screening of clinical S. aureus isolates for novel AIP groups revealed a variant that differed from the group I AIP by a single amino acid residue (aspartate to tyrosine) in the same position defined as critical by alanine scanning. Although this AIP inhibits group I S. aureus strains, the producer strains possess a functional agr locus dependent on the endogenous peptide and, as such, constitute a fourth S. aureus AIP pheromone group (group IV). The addition of exogenous synthetic AIPs to S. aureus inhibited the production of toxic shock syndrome toxin (TSST-1) and enterotoxin C3, confirming the potential of quorum-sensing blockade as a therapeutic strategy. PMID- 11489135 TI - Identification and characterization of a family of secretion-related small GTPase encoding genes from the filamentous fungus Aspergillus niger: a putative SEC4 homologue is not essential for growth. AB - DNA fragments containing genetic information for five secretion-related small GTPases of Aspergillus niger (srgA-E) were isolated and identified as members of different Rab/Ypt subfamilies. This isolation and the search for similar sequences in fungal genomic and EST databases showed that, in contrast to Saccharomyces cerevisiae, filamentous fungi also possess homologues of mammalian Rab2 GTPases. Multiple transcripts with unusually long 5' and 3' untranslated regions were found for all srg genes. Their level of expression was independent of the type of carbon source used for growth. Although the transcripts of srgA and srgB were abundant to the same extent throughout the cultivation, that of the other genes peaked during the early growth phase and then declined. Two genes, srgA and srgB, were characterized further. The protein encoded by srgA exhibited relatively low identity (58%) to its closest S. cerevisiae homologue SEC4, whereas the protein encoded by srgB showed 73% identity with S. cerevisiae YPT1. In contrast to other SEC4 homologues, srgA was unable to complement an S. cerevisiae sec4 mutant, and its disruption was not lethal in A. niger. SrgA mutants displayed a twofold increase in their hyphal diameter, unusual apical branching and strongly reduced protein secretion during growth on glucose. PMID- 11489136 TI - The ubiquitin protein catabolic disorders. AB - The ubiquitin-proteasome system of intracellular proteolysis is essential for cell viability. We propose the concept that neurodegenerative diseases such as Alzheimer's and Parkinson's, as well as other conditions including some types of cancer, collectively represent a raft of 'ubiquitin protein catabolic disorders' in which altered function of the ubiquitin-proteasome system can cause or directly contribute to disease pathogenesis. Genetic abnormalities within the ubiquitin pathway, either in ubiquitin-ligase (E3) enzymes or in deubiquitinating enzymes, cause disease because of problems associated with substrate recognition or supply of free ubiquitin, respectively. In some cases, mutations in protein substrates of the ubiquitin-proteasome system may directly contribute to disease progression because of inefficient substrate recognition. Mutations in transcripts for the ubiquitin protein itself (as a result of 'molecular misreading') also affect ubiquitin-dependent proteolysis with catastrophic consequences. This has been shown in Alzheimer's disease and could apply to other age-associated neurodegenerative conditions. Within the nervous system, accumulation of unwanted proteins as a result of defective ubiquitin-dependent proteolysis may contribute to aggregation events, which underlie the pathogenesis of several major human neurodegenerative diseases. PMID- 11489137 TI - The relationship between alphaB-crystallin and neurofibrillary tangles in Alzheimer's disease. AB - AlphaB-crystallin is known as a small heat shock protein with a cytoprotective function. This study was undertaken to investigate the relationship between alphaB-crystallin and changes seen in Alzheimer's disease. The distribution and immunohistochemical characteristics of alphaB-crystallin positive neurones in the cerebral cortices of 4 patients with Alzheimer's disease were examined. AlphaB crystallin positive neurones were mainly distributed in the limbic and paralimbic regions, namely parahippocampal gyrus, fusiform gyrus, cingulate cortex, middle and superior frontal gyrus, and insular cortex, which corresponded to commonly affected regions in Alzheimer's disease. Moreover, such neurones were present predominantly in the III or V layer of the cerebral cortex. The number of alphaB crystallin positive neurones increased in parallel with the neuronal loss. Logistic regression analysis revealed a significant relation between the density of alphaB-crystallin positive neurones and that of extracellular neurofibrillary tangles (NFTs), with a correlation coefficient (r) of 0.57 and P < 0.0001 in 14 regions of the cerebral cortex. In contrast, the relation was not statistically significant between the density of alphaB-crystallin positive neurones and that of classical senile plaques, diffuse plaques or intracellular NFTs. Modified Gallyas-Braak (GB) staining on alphaB-crystallin positive neurone demonstrated several patterns of the structures: faint GB positive structures in the swollen perikaryon with straight neurites, fine granules compressed and contorted into fuzzy bundles, intensely GB positive filamentous structures together with fine granules and very intensely GB positive ring-like NFTs in a swollen perikaryon with curved neurites. In positive neurones, the density of ring-like NFTs correlated with that of atrophic perikaryon, or bent neurites and a decrease in the immunoreactivity of alphaB-crystallin. These data suggest that a close relationship exists between the appearance of alphaB-crystallin in neurones, extracellular NFTs, and neurofibrillary formation in alphaB-crystallin positive neurones in Alzheimer brain. PMID- 11489138 TI - Cases of Alzheimer's disease due to deletion of exon 9 of the presenilin-1 gene show an unusual but characteristic beta-amyloid pathology known as 'cotton wool' plaques. AB - The pattern of deposition of amyloid beta protein (Abeta) was investigated, using the monoclonal antibodies BA27 and BC05 detecting the C-terminal species Abeta40 and Abeta42(43), in six cases of Alzheimer's disease (AD) due to deletions in exon 9 of PS-1 gene. These cases are characterized histologically by the presence of very large rounded plaques within the frontal cortex, known as 'cotton wool' plaques, composed of both Abeta40 and Abeta42(43) that are relatively free from neuritic changes and glial cell components, and usually devoid of a compact amyloid core. In the cerebellum the plaques are almost entirely of a compact type, again composed of Abeta40 and Abeta42(43), with only few diffuse Abeta42(43) containing plaques. The area fraction of Abeta40, and the ratio between Abeta40 and Abeta42(43), in frontal cortex was significantly higher than that seen in other cases of AD due to different PS-1 mutations, or in cases of sporadic AD, all of similar APO E genotype. The area fractions of Abeta42(43), however, did not significantly differ between these three groups. The unusual nature of the Abeta deposition in these cases may reflect the uniqueness of the mutation, which results in a failure to constitutively cleave the PS-1 holoprotein into its active form, and the effect this might have on APP trafficking and catabolism. PMID- 11489139 TI - Tau-associated neuropathology in ganglion cell tumours increases with patient age but appears unrelated to ApoE genotype. AB - Ganglion cell tumours, including gangliogliomas and gangliocytomas, are low grade neoplasms with a mature neuronal component. Ganglion cells within these lesions occasionally exhibit neurodegenerative changes including neurofibrillary tangles (NFT) similar to those in Alzheimer's disease. The frequency and spectrum of degenerative pathology in these lesions have not been defined, nor has their relation to patient age or factors such as apolipoprotein E (ApoE) genotype that predispose to Alzheimer's disease. We studied 72 ganglion cell tumours (61 gangliogliomas, 11 gangliocytomas) from patients 7 months to 72-years-old. Haematoxylin and eosin (H&E), silver stains (Hirano method) and immunohistochemistry for tau, alpha-synuclein and beta-amyloid were performed on formalin-fixed, paraffin-embedded tissue from surgical specimens. Tau-and silver positive NFT and neuropil threads (NPT) were present in four of 26 ganglion cell tumours from patients over 30-years-old (ages 31, 38, 50, and 58 years). Neuronal granulovacuolar degeneration (GVD) was noted in five of 26 tumours from patients over 30-years-old (mean, 48 years). NFT, NPT, and GVD were not seen in ganglion cell tumours from patients under 30-years-old[0/46]. Cytoplasmic argentophilic bodies distinct from NFT were present in five of 26 tumours from patients over 30 years-old and in two of 46 under 30 years. Neither alpha-synuclein positive neuronal inclusions nor beta-amyloid immunoreactivity was noted in ganglion cell tumours from any age group. The distribution of ApoE genotypes was similar among those tumours that contained tau-associated neuropathology and those that did not. Neurodegenerative changes are uncommon in ganglion cell tumours, but increase in frequency with patient age. GVD, tau-positive NFT and NPT, and argentophilic bodies occur more often in ganglion cell tumours from patients over 30-yrs-old, but do not appear to be associated with a specific ApoE genotype. PMID- 11489140 TI - Accelerated ageing changes in the choroid plexus of a case with multiple mitochondrial DNA deletions. AB - Mitochondrial abnormalities, in particular the accumulation of mitochondrial DNA mutations, have been proposed as a potential cause of normal ageing. One group of patients with mtDNA disorders have a nuclear DNA defect which accelerates the chronological accumulation of mitochondrial DNA mutations. These patients provide an ideal means of investigating whether accelerated mitochondrial DNA defects can cause accelerated ageing pathology. The choroid plexus demonstrates a robust accumulation of pathological changes, in the form of Biondi bodies, with normal ageing. We have therefore examined the choroid plexus of a case with multiple mitochondrial DNA deletions for evidence of accelerated ageing and compared it with two cases with point mutation mitochondrial DNA disorders and several age matched and elderly controls with and without clinical and neuropathological evidence of neurodegenerative disease. We also demonstrate that the choroid plexus of the mitochondrial DNA cases contain cells with levels of mitochondrial DNA mutation sufficient to cause a biochemical deficiency in the oxidative phosphorylation pathway. As previously reported, both cases with point mutation mitochondrial DNA disorders exhibit a characteristic oncocytic type transformation of the choroidal epithelial cells. However, in the case with multiple mitochondrial DNA deletions we demonstrate pathological changes in choroid plexus that are strongly suggestive of accelerated ageing. We believe that this finding supports the theory that the accumulation of mitochondrial DNA mutations can lead to pathological changes typical of ageing cells. PMID- 11489141 TI - Gene expression of adrenomedullin, leptin, their receptors and neuropeptide Y in hormone-secreting and non-functioning pituitary adenomas, meningiomas and malignant intracranial tumours in humans. AB - The aim of this study was to assess human intracranial tumours for their gene expression pattern of the vasoactive peptide adrenomedullin (AM), its receptor (AM-R) and leptin, which exerts multiple biological effects including proliferation and angiogenesis via the leptin receptor (OB-Rb). Gene activity of neuropeptide Y (NPY) was monitored additionally. We investigated whether there was a characteristic gene expression pattern of AM and leptin in different intracranial tumours, depending on their proliferation activity and biological behaviour. We investigated 35 non-functioning pituitary adenomas (including eight null cell, four silent plurihormonal, 23 silent gonadotroph adenomas), seven somatotropinomas, seven prolactinomas, eight meningiomas, five astrocytomas, two glioblastoma multiformes and unaffected temporal lobe (n = 8). Quantitative reverse transcriptase-polymerase chain reaction (TaqMan RT-PCR) was performed. AM mRNA was detectable in all tumour specimens. AM/GAPDH (glyceraldehyde-3-phosphate dehydrogenase) ratio was significantly higher in somatotropinomas, as was AM/CD31 ratio in prolactinomas, compared with inactive adenomas (P < 0.05). AM-R mRNA was found in all tumour subgroups in small quantities but, in general, higher in tumours than in temporal lobe tissue, respectively. AM-R/CD31 ratio was significantly higher in prolactinomas than in inactive adenomas (P < 0.05). Leptin was detectable in very low quantities in each subgroup. OB-Rb gene expression was found in all tumour subgroups, OB-Rb/GAPDH ratio was highest for meningiomas (P < 0.0001, compared with temporal lobe). NPY mRNA was detectable in temporal lobe in higher quantities than in tumours (P < 0.0001), and almost undetectable in prolactinomas and astrocytomas. Our data demonstrate that AM and AM-R, NPY, as well as leptin and OB-Rb, are expressed in various intracranial tumours in humans but their particular function has to be elucidated further. At present, there is no evidence for a cross-talk on transcriptional level between the peptidergic vasodilative system AM and the putative angiogenic and proliferation affecting factor leptin. PMID- 11489142 TI - Ionotropic and metabotropic glutamate receptor protein expression in glioneuronal tumours from patients with intractable epilepsy. AB - Glioneuronal tumours are an increasingly recognized cause of chronic pharmaco resistant epilepsy. In the present study the immunocytochemical expression of various glutamate receptor (GluR) subtypes was investigated in 41 gangliogliomas (GG) and 16 dysembryoplastic neuroepithelial tumours (DNT) from patients with intractable epilepsy. Immunocytochemistry with antibodies specific for ionotropic NR1, NR2A/B (NMDA) GluR1, GluR2 (AMPA), GluR5-7 (kainate), and metabotropic mGluR1, mGluR2-3, mGluR5, mGluR7a subtypes demonstrated in both GG and DNT the presence of an highly differentiated neuronal population, containing subunits from each receptor class. More than 50% of tumours contained a high percentage of neuronal cells immunolabelled for NMDA, AMPA and kainate receptor subunits. A high percentage of neurones showed strong expression of NR2A-B, which co localized with NR1. Group I mGluRs (mGluR1 and mGluR5) were highly represented in the neuronal component of the tumours. Immunolabelling for several GluRs was also present in the glial component. Increased expression of mGluR2-3, mGluR5 and GluR5-7 was observed in reactive astrocytes in the perilesional zone compared to normal cortex. The neurochemical profile of glioneuronal tumours, with high expression of specific GluR subtypes, supports the central role of glutamatergic transmission in the mechanisms underlying the intrinsic and high epileptogenicity of these lesions. PMID- 11489143 TI - Trigeminal neuralgia due to multiple sclerosis: ultrastructural findings in trigeminal rhizotomy specimens. AB - Trigeminal neuralgia is a well-recognized complication of multiple sclerosis. In patients with neuralgia not responding to medical treatment or transcutaneous ablative procedures, the pain can often be treated successfully by partial rhizotomy of the trigeminal sensory root. We have examined partial trigeminal rhizotomy specimens from six multiple sclerosis patients, aged between 34 and 77 years, with intractable trigeminal neuralgia lasting between 18 months and 11 years. The rhizotomy specimens were placed in buffered glutaraldehyde immediately after resection, and subsequently processed for electron microscopy. In all cases, this revealed demyelination in the proximal (CNS) part of the nerve root, with associated gliosis and variable inflammation. A consistent feature was the presence of clusters of juxtaposed axons without intervening glial processes. Similar juxtaposition of axons was previously observed in trigeminal neuralgia due to vascular compression of the nerve root. Experimental studies indicate that this arrangement of demyelinated axons is conducive to both spontaneous impulse activity and ephaptic spread of excitation. The demyelination and associated juxtaposition of axons may therefore account for key aspects of the pathogenesis of trigeminal neuralgia. PMID- 11489144 TI - Bilateral hippocampal granule cell dispersion: autopsy study of 3 infants. AB - Recent morphologic studies of Ammon's horn sclerosis (AHS) have recorded granule cell dispersion (GCD) in a significant proportion of temporal lobes surgically resected for temporal lobe epilepsy (TLE). We report the first post-mortem descriptions of GCD in three unrelated infants. GCD was bilateral in all three and there were also migrational defects, heterotopias or polymicrogyria. Only one child, a 2.5-year-old boy, presented with a severe seizural disorder and exhibited bilateral AHS. But in the two younger children, deceased at 12 weeks and 5 months, respectively, no seizures were observed and hippocampal neuronal populations were intact. To date, GCD has only been reported in association with epilepsy and has not been observed bilaterally or in neurologically normal individuals. The present observations bring into question the hypothesis that GCD is causally related to seizure activity in early life, suggesting an opposing view that it is an independent developmental disorder. PMID- 11489145 TI - The difficult problems of authorship in regard to large longitudinal data sets. PMID- 11489146 TI - Myopic rescuers. PMID- 11489147 TI - Birthweight, vitamin D receptor genotype and the programming of osteoporosis. AB - Studies of the association between polymorphisms of the gene for the vitamin D receptor (VDR) and adult bone mass have been inconsistent, pointing to the possibility that gene--environment interactions may vary in different populations. We have demonstrated previously an association between weight in infancy (a marker of the intrauterine and early post-natal environment) and each of adult bone mass and VDR genotype. We therefore sought to extend these observations in an elderly UK cohort and to investigate the possibility of an interaction between these genetic and early environmental markers of later osteoporosis risk. One hundred and sixty-five men and 126 women aged 61--73 years for whom birth records were available underwent bone mass measurements at baseline and follow-up 4 years later. Whole-blood samples were obtained, DNA extracted using standard techniques and polymorphisms in the VDR and collagen type I alpha 1 (Col IA1) genes identified. In the cohort as a whole, there were no significant associations between either birthweight or VDR genotype and bone mineral density (BMD) or bone loss rate at either site. However, the relationship between lumbar spine BMD and VDR genotype varied according to birthweight. Among individuals in the lowest third of birthweight, spine BMD was higher (P = 0.01) in individuals of genotype 'BB' after adjustment for age, sex and weight at baseline. In contrast, spine BMD was reduced (P = 0.04) in individuals of the same genotype who were in the highest third of the birthweight distribution. A significant (P = 0.02) statistical interaction was also found between VDR genotype and birthweight as determinants of BMD. Similar but slightly weaker associations were seen between lumbar spine bone mineral content (BMC) and VDR genotype in the lowest birthweight tertile. When examining the relationship between Col1A1 genotype and bone mass, lumbar spine BMC was higher in individuals of genotype 'Ss' or 'ss' in the lowest birthweight tertile (P = 0.02) after adjustment for age, sex and weight at baseline. These results suggest that genetic influences on adult bone size and mineral density may be modified by undernutrition in utero. PMID- 11489148 TI - Adult survival after prenatal exposure to the Dutch famine 1944--45. AB - Early life events may affect adult survival. We studied the effect of prenatal exposure to the Dutch famine 1944--45 on survival among 2254 people born in Amsterdam. Mortality up to age 50 was highest among those born before the famine (15.2%) and among those exposed to famine in late gestation (14.6%). It was lower among those exposed in mid- (11.2%) or early gestation (11.5%), and was lowest among those conceived after the famine (7.2%). These differences were caused by effects on mortality in the first year after birth and were mainly related to nutrition and infections. There was no effect of exposure to famine on mortality after the age of 18. The hazard ratio was 1.4 [0.8, 2.3] for those born before the famine, 1.1 [0.5, 2.3] for those exposed in late gestation, 0.8 [0.3, 1.8] for those exposed in mid-gestation and 1.1 [0.5, 2.5] in those exposed in early gestation compared with those conceived after the famine. We could not demonstrate effects of prenatal exposure to famine on cause-specific mortality after the age of 18. Because prenatal exposure to famine is linked to cardiovascular risk factors and disease, increased cardiovascular mortality in the future may be expected. PMID- 11489149 TI - Risks of hypertensive disorders in the second pregnancy. AB - This study examined the incidence of and risk factors for recurrent and newly developed hypertensive disorders in the second pregnancy. We analysed data on 1641 women who had both the first and second pregnancies in the Collaborative Perinatal Project, a large prospective cohort study at 12 hospitals in the US. Nineteen per cent [95% CI 14%, 24%] of women who had gestational hypertension in the first pregnancy, 32% [95% CI 17%, 48%] of those with pre-eclampsia and 46% [95% CI 32%, 60%] of patients with gestational hypertension or pre-eclampsia superimposed on chronic hypertension, had recurrent hypertensive disorders in the second pregnancy. Risk factors for recurrence included history of chronic hypertension and thromboembolism, early onset of hypertension in the first pregnancy or persistent hypertension after 5 weeks postpartum and high baseline blood pressure in the second pregnancy. Women with a normotensive first pregnancy but a severe small-for-gestational-age birth had twice the risk of developing hypertension in the second pregnancy (RR = 2.1, 95% CI, 1.1, 4.0). In summary, hypertensive disorders have a 20--50% recurrence rate in the second pregnancy. The earlier the onset of hypertension in the first pregnancy, the higher the overall recurrence rate. Intrauterine growth restriction of the first birth is an independent risk factor for hypertension in the second pregnancy. PMID- 11489150 TI - Delivery method and self-reported postpartum general health status among primiparous women. AB - Despite nearly four million deliveries in the United States each year, minimal information exists on unintended health consequences following childbirth, particularly in relation to delivery method. The purpose of this study was to assess the association between method of delivery and the general health status, sexual, bowel and urinary functioning of primiparous women as measured at 7 weeks postpartum. Data from the Statewide Obstetrical Review of Quality System (StORQS) Survey of Maternity Care in Washington State were analysed. Participants included all primiparous women with a delivery of a singleton infant discharged alive between August and December 1991 from 10 non-federal short-stay hospitals who responded to the StORQS Survey of Maternity Care (n = 971). The main outcome measures included the modified Medical Outcomes Study 36-Item Short-Form Health Survey and self-reported sexual, bowel and urinary functioning. At 7 weeks postpartum, women who had caesarean or assisted vaginal deliveries reported significantly lower postpartum general health status scores than women with unassisted vaginal delivery. Additionally, women with assisted vaginal delivery reported significantly worse sexual, bowel and urinary functioning. Our results suggest that more careful attention to the postpartum general health and sexual functioning of women with caesarean and assisted vaginal delivery may be merited. PMID- 11489151 TI - Vaginal delivery vs. a 'nice clean cut': giving more attention to mothers' postpartum health. PMID- 11489152 TI - Tocolytic therapy for preterm labour: assessing its potential for reducing preterm delivery. AB - Preterm labour (PTL) is a major contributor to preterm delivery (PTD) but delivery is often not preventable by current therapies. We conducted this study to determine the proportion of women with PTL who were and who were not candidates for tocolytic therapy. The cohort comprised residents of Olmsted County, Minnesota who delivered at >20 weeks' gestation in 1985--94 and who experienced PTL. Medical records were abstracted to identify episodes of PTL, its treatment and outcome. We developed an algorithm that accounted for gestation at delivery and pregnancy complications to determine the proportion of pregnancies complicated by PTL that were candidates for tocolytic therapy. Of 651 pregnancies complicated by PTL, a 50% probability sample, stratified by delivery year, were selected and abstracted. The cumulative incidence of PTL ranged from 3.6 to 6.4 per 100 deliveries of live or stillborn infants. Tocolysis was not contraindicated for 49.4% of all women with PTL and for a third of women with only one PTL episode. Delivery was delayed to >35 weeks in 53.8% of candidates for tocolysis. Only an additional 11.7% of women with one or more PTL episodes could have had their PTD delayed beyond 35 weeks if a perfect tocolytic therapy had been available. Many pregnancies complicated by PTL occurred at > or =35 weeks or involved maternal or obstetric factors that contraindicated tocolytic medications. The maximum incremental benefit that could be expected of a new safe and efficacious tocolytic therapy would be to reduce current PTD rates resulting from PTL by about 12%. PMID- 11489153 TI - Preterm labour at 34--36 weeks of gestation: should it be arrested? AB - Currently, preterm labour is treated with tocolytic agents and prenatal steroids until the 34th week of gestation only. Our objective in this study was to assess this practice. Seven-year records of all preterm infants born in our institution at 34--36 weeks of gestation, were evaluated retrospectively. All babies, born in singleton well-dated pregnancies, without maternal, medical or obstetric complications, and by normal vaginal delivery, were included. Their length of hospital stay and perinatal complications were compared across gestational age groups of 34, 35 and 36 weeks. Of the 207 babies included, statistically significant reductions in the rates of respiratory distress syndrome (15.0% vs. 3.2%), nosocomial sepsis (5.0% vs. 0%) and apnoea of prematurity (11.7% vs. 2.2%), and consequently, in length of hospital stay (16 +/- 2.7 vs. 4 +/- 0.3 days) occurred between 34 and 36 weeks of gestation. The severity of respiratory distress syndrome also declined significantly. The changes were most noticeable after 35 weeks of gestation, and it was concluded that neonatal complications are still prevalent at 34 and 35 weeks. Therefore, we propose that labour should not be induced at 34 and 35 weeks of gestation and that tocolytic agents and maternal prenatal steroids may be considered in preterm labour during this period. PMID- 11489154 TI - Risk factors for low birthweight in north-east Brazil: the role of caesarean section. AB - Caesarean section (CS) delivery is associated with low birthweight (LBW) in south east Brazil. A hospital-based study was conducted on singleton infants from mothers residing in Sao Luis, to assess if an association between CS and LBW was found in the northern part of the country, where the CS rate is lower than in the south-east. A standardised questionnaire was administered to a sample of 2541 mothers in 10 hospitals, representing 94% of all deliveries, from March 1997 to February 1998. In a logistic model, type of delivery was the independent variable, the other variables were treated as confounders, and interaction terms were added between type of delivery and all other factors. LBW was associated with low maternal height, maternal smoking, primiparity, previous LBW, public insurance, preterm birth and CS. The CS rate was 33.7%. The risk of CS was higher for primiparous and married mothers, those with high level of schooling and attended by the same physician during prenatal and delivery care, deliveries held in private hospitals, daylight hours or evenings, and for those mothers who had adequate prenatal care. Because it appears unlikely that only medical reasons are operative, it is a possibility that CS could cause LBW, reflecting abusive indications for elective CS. PMID- 11489155 TI - The efficacy of the non-stress test in preventing fetal death in post-term pregnancy. AB - We conducted a case--control study to examine the efficacy of non-stress testing in preventing fetal death in post-term pregnancy. The analysis was based on data from the 1988 National Maternal and Infant Health Survey, which was a nationally representative sample of live births, fetal deaths and infant deaths that occurred in 1988. Information on whether a woman had non-stress testing was obtained from a questionnaire sent to prenatal care providers and hospitals. Cases were post-term women (with 42 weeks or more gestation) who had fetal deaths. Three post-term controls, who had live births and who delivered at the same time or later than the cases, were randomly chosen and individually matched to each case by maternal race. The proportion of women who had one or more non stress tests during pregnancy was compared between cases and controls. Non-stress testing was used in 30.9% of the 126 cases and in 28.5% of the 375 controls. The race-adjusted odds ratio for exposure to non-stress test was 1.12 [95% CI 0.72, 1.75]. After controlling for other important confounding variables the odds ratio was 1.05 [95% CI 0.57, 1.91]. These results do not support the efficacy of non stress testing in post-term pregnancies. A more detailed evaluation of this widely used screening procedure is needed. PMID- 11489156 TI - Cerebral palsy births in eastern Denmark, 1987--90: implications for neonatal care. AB - The Cerebral Palsy Register in eastern Denmark has collected cases using a uniform data sampling procedure since birth year 1979. We have investigated changes in the rate of cerebral palsy, related to gestational age, mortality and perinatal risk factors in children born 1983--90. The total cerebral palsy birth prevalence decreased from 3.0 in the birth year period 1983--86 to 2.4 per 1000 live births (P < 0.01) in 1987--90, owing to a decrease among all preterm infants (29--19 per 1000, P < 0.001). The perinatal and early neonatal mortality in preterm infants was unchanged from 1983--86 to 1987--90. The rate of cerebral palsy in term infants was 1.5 per 1000 in all birth-year periods from 1979--90. Among the cerebral palsy infants, the proportion of very preterm babies treated with mechanical ventilation in the neonatal period decreased from 95% in 1983--86 to 61% in 1987--90 (P < 0.001), while the group treated with CPAP among the moderately preterm babies increased from 61% to 78% (P < 0.05). The significant decline in cerebral palsy rate in preterm infants born 1987--90 may be due to a change in treatment at the neonatal intensive care units using less mechanical ventilation, a hypothesis which needs further investigation. PMID- 11489157 TI - Cerebral palsy in Northern Ireland: 1981--93. AB - This paper describes the method of compilation of the Northern Ireland Cerebral Palsy Register (NICPR) and outlines the epidemiology of cerebral palsy (CP) in Northern Ireland in 1981--93 based on an analysis of 784 cases. Multiple and overlapping sources of notification were used to compile the NICPR. Each case was followed up with a standardised assessment completed at age 5 years and validated by a paediatrician. Over 70% of cases were multiply notified, and paediatricians were the most important source of notification. Cases of CP most likely to be under-ascertained included those very mildly affected, cases who died before the register began (death certificates were searched but found to be unreliable) and cases who moved out of the area before their diagnosis was confirmed. The prevalence of CP for the birth period 1981--93 was 2.24 per 1000 livebirths [95% CI 2.08, 2.40]. There were no statistically significant temporal or geographic variations in the rate of CP. Almost half the cases of CP were of low birthweight (<2500 g), and this proportion increased during 1987--89, but decreased (P < 0.01) more recently (1990--93). The proportion of cases weighing <1000 g trebled from 3% in 1981--83 to 10% in 1990--93 (P < 0.01). The most common CP subtype was bilateral spastic cerebral palsy (55%). Just over a quarter (29%) of cases were unable to walk (with/without aids), and one-fifth (22%) had no useful hand/arm function. Almost half (49%) the cases had at least one other impairment (intellectual, sensory impairment or active seizures) in association with their CP. Prevalence rates and the pattern of disability in populations of people with CP can only be evaluated on the basis of the methods of case definition and ascertainment. We have demonstrated the methods necessary to ensure valid, standard and detailed information on CP for the purposes of surveillance, service planning and research. PMID- 11489158 TI - Atmospheric pressure and sudden infant death syndrome in Cook County, Chicago. AB - We investigated daily deaths from sudden infant death syndrome (SIDS) in Cook County, Chicago, from 1 January 1986 to 31 December 1990. The daily SIDS rate was 0.54 deaths/day. Mean daily atmospheric pressure ranged from 952 to 1007 mb. We found a weak link with barometric pressure, whereby an increase of 10 mb was associated with an increased risk of 12% [95% CI 0.1%, 24%]. Cross-tabulation revealed that the highest SIDS rates were associated with a drop from high to low pressure. Possible reasons for this are discussed. PMID- 11489159 TI - Placental abruption and perinatal death. AB - Studies of risk factors for abruptio placentae (AP) are partly conflicting and studies of risk factors for perinatal death in these pregnancies are scarce. Using the population-based Swedish Birth Registry from 1987 to 1993, we were able to study these risks in 795,459 singleton pregnancies. Logistic regression analysis was used to estimate odds ratios (OR) for risk of AP and risk of perinatal death in pregnancies with and without AP. Risk factors for AP were: age, primiparity, high parity, not cohabiting with infant's father, low education, smoking, infertility, pregestational diabetes, essential hypertension, pregnancy-induced hypertensive diseases, preterm premature rupture of membranes, preterm birth and small-for-gestational-age (SGA) births. Risk factors for perinatal death in pregnancies with placental abruption were smoking (1--9 and > or =10 cigarettes/day; OR 1.4 and 1.7 respectively), severe pre-eclampsia (OR 2.0) and SGA (OR 1.9), whereas in pregnancies without abruption, risks were also increased in maternal age > or =35 years, primiparity, infertility, essential hypertension and pregestational diabetes. These findings support the theory that, in cases of AP, a general impairment of the placenta and/or a defect placentation may be fatal. PMID- 11489160 TI - Perinatal mortality in twins and factors influencing mortality in Japan, 1980- 98. AB - The perinatal mortality rate (PMR) in twins was analysed using Japanese vital statistics during the period 1980-98. There was a total of 1992 twin perinatal deaths. The PMR in twins significantly decreased from 91 per 1000 births in 1980 to 31 per 1000 in 1998. The PMR was higher in males than females, but the differences between the sexes decreased over time. It was significantly higher in the second-born than the first-born of the twins. The fetal death rate was 2.8 times higher in monozygotic than dizygotic twins. The PMR decreased to one-sixth for the maternal age group > or =40 years, one-fifth for <20 years, and between one-third and a half for 20--39 years during the period 1980--98. It was lowest for birthweights (BW) between 2500 g and 2999 g during the entire period; the PMR decreased with gestational age (GA) up to 38--39 weeks and increased thereafter. The effect of BW on the PMR was greater than that of GA except in the shorter GA group. The proportion of twin perinatal deaths with extremely low BW (<1000 g) increased from 50% in 1980 to 68% in 1998. Thus, the declining PMR was unlikely to be due to the improvement in BW in twins. It is likely that it is related to the improved medical management of twins during the perinatal period and the first week of life. PMID- 11489161 TI - The reliability of perinatal and neonatal mortality rates: differential under reporting in linked professional registers vs. Dutch civil registers. AB - Official Dutch perinatal mortality rates are based on birth and death certificates. These civil registration data are not detailed enough for international comparisons or extensive epidemiological research. In this study, we linked and extrapolated three national, incomplete, professional registers from midwives, obstetricians and paediatricians, containing detailed perinatal information. This linkage and extrapolation resulted in one detailed professional database which is representative of all Dutch births and from which gestational age-specific perinatal mortality rates could be calculated. The reliability of these calculated mortality rates was established by comparing them with the rates derived from the national civil registers. The professional database reported more perinatal deaths and fewer late neonatal deaths than the civil registers. The under-reporting in the civil registers amounted to 1.2 fewer perinatal deaths per 1000 births and was most apparent in immature newborns. We concluded that under-reporting of perinatal and neonatal deaths depends on the data source used. Mortality rates for the purpose of national and international comparison should, therefore, be defined with caution. This study also demonstrated that combining different incomplete professional registers can result in a more reliable database containing detailed perinatal information. Such databases can be used as the basis for extensive perinatal epidemiological research. PMID- 11489162 TI - Perinatal mortality rates can no longer be used for comparing quality of perinatal health services between countries. PMID- 11489163 TI - The mother-only method (MOM) to detect maternal gene--environment interactions. PMID- 11489164 TI - In-vitro susceptibility of hydatid cysts of Echinococcus granulosus to nitric oxide and the effect of the laminated layer on nitric oxide production. AB - Murine hydatid cysts of Echinococcus granulosus were incubated in vitro in the presence of nitric oxide produced from S-nitroso-N-acetylpenicillamine (SNAP) or interferon-gamma activated peritoneal macrophages. In both situations, evidence of cyst damage and death was observed by microscopy in over 77% of cysts after 3 days, indicating that intact hydatid cysts could be susceptible to a Th1 driven macrophage attack. A crude extract of the laminated layer from cysts was found to be able to reduce the production of nitric oxide from activated macrophages in vitro and in vivo and this may have been due to phagocytosis of laminated layer fragments by the macrophages. The results indicate that, although cysts may be susceptible to the effects of nitric oxide, the laminated layer may be involved in downregulating nitric oxide production. PMID- 11489165 TI - Age-related protective immunity after vaccination with Haemonchus contortus excretory/secretory proteins. AB - Protection against an experimental challenge infection by immunization with excretory/secretory products (ES) from Haemonchus contortus, containing predominantly proteins of 15 and 24 kDa, depends on the age of the sheep. Vaccinated sheep 9 and 6 months of age had reduced final worm burdens of 82 and 77, respectively. No reduction in worm burden was found in 3-month-old lambs. Nine-month-old sheep had significantly higher ES-specific serum immunoglobulin (Ig)G1 and IgA during immunizations and after challenge infection than 3-month old lambs. There was no correlation within the 9-month-old sheep between ES specific IgA or IgG1 levels and protection, measured as worm burden. However, when the different age groups were combined, negative correlations between percentage protection and ES-specific IgA and IgG1 levels after challenge were found. At the end of the experiment, peripheral blood eosinophils and mast cell counts in abomasal tissue were also significantly higher in the vaccinated and challenged 9-month-old sheep than in the vaccinated and challenged 3-month-old or than in the 9-month-old sheep with challenge, but without vaccination. The responses measured in young lambs were similar to the responses in sheep, but the height of these responses was in general of a lower magnitude. PMID- 11489166 TI - A glycoprotein from Schistosoma mansoni eggs binds non-antigen-specific immunoglobulin E and releases interleukin-4 from human basophils. AB - We have recently shown that soluble extracts from Schistosoma mansoni eggs (SmEA) triggered basophils from nonsensitized donors to rapidly release interleukin (IL) 4. Assuming that this mechanism might play a role in vivo in biasing the immune response towards a Th2 phenotype, we determined basic properties of the IL-4 inducing activity contained in SmEA. Sensitivity to pepsin digestion indicated protein nature. Binding to and specific elution from Concanavalin A-sepharose suggested that this protein contains mannose residues, thus being a glycoprotein. The IL-4-inducing activity was stable for 30 min at room temperature towards shifting the pH between 3 and 10. When incubated at 100 degrees C, it was stable at pH 3, but less stable at neutral and alkaline pH. Electroelution from an SDS PAGE gel indicated an apparent molecular weight of the IL-4-inducing activity between 31 and 66 kDa. Although binding to purified human immunoglobulin E (IgE) and activating basophils IgE-dependently, SmEA appears to activate basophils in a non-antigen-specific way, since the cells were purified from noninfected donors. Because the IL-4-inducing activity was found to be released from eggs, it could be an important factor in the environment of the eggs skewing the immune response towards the Th2 phenotype. PMID- 11489167 TI - Immunogenicity of recombinant fragments of Plasmodium falciparum acidic basic repeat antigen produced in Escherichia coli. AB - The acidic basic repeat antigen (ABRA) of Plasmodium falciparum is a potential vaccine candidate against erythrocytic stages of malaria. We report, for the first time, the immunological characteristics of recombinant ABRA constructs. The recombinant proteins representing different fragments of ABRA were expressed in Escherichia coli, either as fusions with maltose binding protein or as 6X histidine tagged molecules, and purified by affinity chromatography. Immunogenicity studies with these constructs in rabbits and mice indicated that the N-terminal region is the least immunogenic part of ABRA. T-cell proliferation experiments in mice immunized with these constructs revealed that the T-cell epitopes were localized in the middle portion of the protein. More importantly, the purified immunoglobulin G specific to middle and C-terminal fragments prevented parasite growth at levels approaching 80-90%. We found that these proteins were also recognized by sera from P. falciparum-infected patients from Rourkela, a malaria endemic zone of India. Our immunogenicity results suggest that potential of ABRA as a vaccine candidate antigen should be investigated further. PMID- 11489168 TI - Susceptibility of tumour necrosis factor-alpha genetically deficient mice to Trypanosoma congolense infection. AB - The TNF-alpha gene on mouse chromosome MMU17 is among the candidates for the trypanosomosis resistance QTL Tir1. Tir1 has the largest effect of those loci so far detected which influence degree of resistance to murine trypanosomosis caused by Trypanosoma congolense infection. We therefore studied the survival to 180 days after challenge with T. congolense of mice that were homozygous and hemizygous with respect to a disruption of the TNF-alpha gene on a > 99% C57BL/6 (resistant) background. We also examined the responses of TNF-alpha hemizygous mice produced by crossing the deletion line with mice of the C57BL/6J strain, and with mice of the susceptible A/J strain. Mice lacking a functional TNF-alpha gene were shown to be highly susceptible to challenge with T. congolense with a median survival time of 37 days. This was comparable to 71 days for control wild-type mice, and 61 and 111 days for mice of the susceptible A/J and resistant C57BL/6J strains, respectively. In mice of the deletion line, the C57BL/6 TNF-alpha allele tended to be dominant to the TNF knockout in terms of resistance. We conclude that TNF-alpha plays an important role in resistance to the effects of T. congolense infection in mice. PMID- 11489169 TI - Cellular immune responses of filaria (Litomosoides sigmodontis) infected BALB/c mice detected on the level of cytokine transcription. AB - Cellular immune responses of BALB/c mice infected with 80 or 160 L3 of Litomosoides sigmodontis were studied over a period of 200 days postinfection (p.i.) by stimulating spleen cells with specific microfilariae and adult antigens and Concanavalin A (Con A). Effects were determined as the level of transcription of cytokine genes [interleukin (IL)-2, interferon (IFN)-gamma, IL-4, IL-5, IL-10, IL-13] employing a semiquantitative reverse transcriptase-polymerase chain reaction technique. Con A stimulation resulted in generally enhanced transcription levels in infected animals. Exposure to filarial antigens stimulated T cells of infected animals dependent on time p.i. There was a general strong response in the early prepatency (24 days p.i.), a temporary almost complete downregulation of cytokine gene transcription except IL-10 towards the end of prepatency (45 days p.i.), and subsequently strong reactions particularly concerning IFN-gamma and IL-13 during patency and postpatency. The dose of infection as well as the mode of antigenic stimulation had generally only small effects on the cytokine gene transcription: following the same type of kinetics, infection with 160 L3 as well as the use of microfilarial antigen generally induced lower levels of cytokine gene transcription compared with infection with 80 L3 and stimulation with female antigen, respectively. PMID- 11489170 TI - An Arabidopsis inositol phospholipid kinase strongly expressed in procambial cells: synthesis of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 in insect cells by 5 phosphorylation of precursors. AB - We have cloned a phosphatidylinositol-4-phosphate 5-kinase (PIP5K) cDNA (AtP5K1) from Arabidopsis thaliana. By the application of cell permeabilization and short term nonequilibrium labelling we show that expression of AtP5K1 in Baculovirus infected insect (Spodoptera frugiperda) cells directs synthesis of PtdIns(4,5)P2 and PtdIns(3,4,5)P3. The same phosphoinositides were produced by isolated whole cell membrane fractions of AtP5K1-expressing insect cells. Their synthesis was not affected by adding defined precursor lipids, that is PtdIns(3)P, PtdIns(4)P, PtdIns(3,4)P2, or PtdIns(4,5)P2, in excess, indicating that substrates for the plant enzyme were not limiting in vivo. Enzymatic dissection of lipid headgroups revealed that AtP5K1-directed synthesis of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 proceeds via 5-phosphorylation of precursors. Analysis of promoter-reporter gene (beta-glucuronidase) fusions in transgenic plants revealed that expression of the AtP5K1 gene is strongest in vascular tissues of leaves, flowers, and roots, namely in cells of the lateral meristem, that is the procambium. Single-cell sampling of sap from flower stem meristem tissue and neighbouring phloem cells, when coupled to reverse transcriptase--polymerase chain reaction, confirmed preferential expression of AtP5K1 in procambial tissue. We hypothesize that AtP5K1, like animal and yeast PIP5K, may be involved in the control of cell proliferation. PMID- 11489171 TI - Overexpression of DWARF4 in the brassinosteroid biosynthetic pathway results in increased vegetative growth and seed yield in Arabidopsis. AB - Plants unable to synthesize or perceive brassinosteroids (BRs) are dwarfs. Arabidopsis dwf4 was shown to be defective in a steroid 22alpha hydroxylase (CYP90B1) step that is the putative rate-limiting step in the BR biosynthetic pathway. To better understand the role of DWF4 in BR biosynthesis, transgenic Arabidopsis plants ectopically overexpressing DWF4 (AOD4) were generated, using the cauliflower mosaic virus 35S promoter, and their phenotypes were characterized. The hypocotyl length of both light- and dark-grown AOD4 seedlings was increased dramatically as compared to wild type. At maturity, inflorescence height increased >35% in AOD4 lines and >14% in tobacco DWF4 overexpressing lines (TOD4), relative to controls. The total number of branches and siliques increased more than twofold in AOD4 plants, leading to a 59% increase in the number of seeds produced. Analysis of endogenous BR levels in dwf4, Ws-2 and AOD4 revealed that dwf4 accumulated the precursors of the 22alpha-hydroxylation steps, whereas overexpression of DWF4 resulted in increased levels of downstream compounds relative to Ws-2, indicative of facilitated metabolic flow through the step. Both the levels of DWF4 transcripts and BR phenotypic effects were progressively increased in dwf4, wild-type and AOD4 plants, respectively. This suggests that it will be possible to control plant growth by engineering DWF4 transcription in plants. PMID- 11489172 TI - Transcriptional activation by the sexual pheromone and wounding: a new gene family from Volvox encoding modular proteins with (hydroxy)proline-rich and metalloproteinase homology domains. AB - The green alga Volvox represents the simplest kind of multicellular organism: it is composed of only two cell types, somatic and reproductive, making it suitable as a model system. The sexual development of males and females of Volvox carteri is triggered by a sex-inducing pheromone at a concentration of < 10-16 M. Early biochemical responses to the pheromone involve structural modifications within the extracellular matrix (ECM). By differential screenings of cDNA libraries made from mRNAs of pheromone-treated Volvox, four novel genes were identified that encode four closely related Volvox metalloproteinases that we use to define a new protein family, the VMPs. The existence of several features common to matrix glycoproteins, such as signal peptides, a (hydroxy)proline content of 12-25%, and Ser(Pro)2-4 repeats, suggest an extracellular localization of the VMPs within the ECM. Synthesis of VMP cDNAs is triggered not only by the sex-inducing pheromone, but also by wounding, and is restricted to the somatic cell type. Sequence comparisons suggest that the VMPs are members of the MB clan of zinc-dependent matrix metalloproteinases, although the putative zinc binding site of all VMPs is QEXXHXXGXXH rather than HEXXHXXGXXH. The presence of glutamine instead of histidine in the zinc binding motif suggests a novel family, or even clan, of peptidases. Like the matrixin family of human collagenases, Volvox VMPs exhibit a modular structure: they possess a metalloproteinase homology domain and a (hydroxy)proline-rich domain, and one of them, VMP4, also has two additional domains. Metalloproteinases seem to be crucial for biochemical modifications of the ECM during development or after wounding in the lower eukaryote Volvox with only two cell types, just as in higher organisms. PMID- 11489173 TI - Involvement of a novel Arabidopsis phospholipase D, AtPLDdelta, in dehydration inducible accumulation of phosphatidic acid in stress signalling. AB - Phospholipid metabolism is involved in plant responses to drought and salinity stress. To investigate the role of phospholipase D (PLD) and its product phosphatidic acid (PtdOH) in stress signalling, we isolated a novel PLD cDNA, designated AtPLDdelta, by screening a cDNA library prepared from dehydrated Arabidopsis thaliana. The AtPLDdelta protein, of 868 amino acids, has a putative catalytic domain and a C2 domain that is involved in Ca2+/phospholipid binding. The AtPLDdelta mRNA accumulated in response to dehydration and high salt stress. Histochemical analysis showed that the AtPLDdelta gene is strongly expressed in the vascular tissues of cotyledons and leaves under dehydration stress conditions. Under normal growth conditions, AtPLDdelta was expressed in roots, leaves, stems and flowers but not in siliques. We showed that dehydration stimulates the accumulation of PtdOH. The accumulation of PtdOH in response to dehydration was significantly suppressed in AtPLDdelta antisense transgenic plants. These results suggest that AtPLDdelta may be involved in PtdOH accumulation in the dehydration stress response. PMID- 11489174 TI - Light regulation of circadian clock-controlled gene expression in rice. AB - Using transgenic rice seedlings expressing a firefly luciferase (luc) gene under the control of a rice CAB (chlorophyll a/b binding protein) promoter, we demonstrated how light affects circadian clocks in the transcription of the CAB gene. Rhythmic luc expression was observed for more than 5 d under constant light and dark (DD) conditions after light/dark entrainment. After a light pulse was applied at different time points in DD various temporal patterns of CAB gene expression were individually observed. We first examined two distinct properties related to the entrainment mechanism of the circadian clock: fluence-rate dependence of free-running periods (FRPs) and phase resetting by a light pulse. Although fluence-rate dependent shortening of FRP was demonstrated, the FRP in DD was almost equal to that in constant light of a middle fluence-rate, indicating that this fluence-rate dependence may not fully describe the entrainment of the circadian clock in rice. Typical phase responses of the circadian clock by a single light pulse were also observed at the transcriptional level in rice seedlings. Thus, the phase resettings upon the light/dark transitions of daily cycles may be sufficient to explain the entrainment mechanisms of rice. We have further demonstrated that, in addition to having a gating effect to acute response, a light pulse can activate the circadian clock-controlled CAB1R gene expression at the first circadian peak in a phase-dependent manner. This suggests that light activates circadian clock activity in the diurnal CAB gene expression under daily light/dark cycles. PMID- 11489175 TI - Gene-associated CpG islands in plants as revealed by analyses of genomic sequences. AB - We screened plant genome sequences, primarily from rice and Arabidopsis thaliana, for CpG islands, and identified DNA segments rich in CpG dinucleotides within these sequences. These CpG-rich clusters appeared in the analysed sequences as discrete peaks and occurred at the frequencies of one per 4.7 kb in rice and one per 4.0 kb in A. thaliana. In rice and A. thaliana, most of the CpG-rich clusters were associated with genes, which suggests that these clusters are useful landmarks in genome sequences for identifying genes in plants with small genomes. In contrast, in plants with larger genomes, only a few of the clusters were associated with genes. These plant CpG-rich clusters satisfied the criteria used for identifying human CpG islands, which suggests that these CpG clusters may be regarded as plant CpG islands. The position of each island relative to the 5'-end of its associated gene varied considerably. Genes in the analysed sequences were grouped into five classes according to the position of the CpG islands within their associated genes. A large proportion of the genes belonged to one of two classes, in which a CpG island occurred near the 5'-end of the gene or covered the whole gene region. The position of a plant CpG island within its associated gene appeared to be related to the extent of tissue-specific expression of the gene; the CpG islands of most of the widely expressed rice genes occurred near the 5'-end of the genes. PMID- 11489176 TI - Physical interactions between ABA response loci of Arabidopsis. AB - Genetic and physiological studies have shown that the Arabidopsis thaliana abscisic acid-insensitive (ABI) loci interact to regulate seed-specific and/or ABA-inducible gene expression. We have used the yeast two-hybrid assay to determine whether any of these genetic interactions reflect direct physical interactions. By this criterion, only ABI3 and ABI5 physically interact with each other, and ABI5 can form homodimers. The B1 domain of ABI3 is essential for this interaction; this is the first specific function ascribed to this domain of the ABI3/VP1 family. The ABI5 domains required for interaction with ABI3 include two conserved charged domains in the amino-terminal half of the protein. An additional conserved charged domain appears to have intrinsic transcription activation function in this assay. Yeast one-hybrid assays with a lacZ reporter gene under control of the late embryogenesis-abundant AtEm6 promoter show that only ABI5 binds directly to this promoter fragment. PMID- 11489177 TI - The protein kinases AtMAP3Kepsilon1 and BnMAP3Kepsilon1 are functional homologues of S. pombe cdc7p and may be involved in cell division. AB - We identified an Arabidopsis thaliana gene, AtMAP3Kepsilon1, and a Brassica napus cDNA, BnMAP3Kepsilon1, encoding functional protein serine/threonine kinases closely related to cdc7p and Cdc15p from Schizosaccharomyces pombe and Saccharomyces cerevisiae, respectively. This is the first report of cdc7-related genes in non-fungal eukaryotes; no such genes have as yet been identified in Metazoans. The B. napus protein is able to partially complement a cdc7 loss of function mutation in S. pombe. RT-PCR and in situ hybridisation revealed that the A. thaliana and B. napus genes are expressed in both the sporophytic and the gametophytic tissues of the respective plant species and revealed further that expression is highest in dividing cells. Moreover, AtMAP3Kepsilon1 gene expression is cell cycle-regulated, with higher expression in G2-M phases. Our results strongly suggest that the plant cdc7p-related protein kinases are involved in a signal transduction pathway similar to the SIN pathway, which positively regulates cytokinesis in S. pombe. PMID- 11489178 TI - Cold-activation of Brassica napus BN115 promoter is mediated by structural changes in membranes and cytoskeleton, and requires Ca2+ influx. AB - Previous studies on cold-triggered events leading to Ca2+ influx during cold acclimatization have been conducted on either unicellular cyanobacterium Synechocystis or plant cell suspensions, and used transcript levels of cold induced genes as end-point markers. Whether the results of these studies are valid for intact plants or their organs is not known. Here we examine cold signaling in transgenic Brassica napus seedlings carrying, in addition to the endogenous cold-inducible BN115 gene, the beta-glucuronidase (GUS) gene placed under control of the BN115 promoter. The activity of BN115 promoter was monitored at the transcriptional and translational levels by determining accumulation of BN115 transcripts and by histochemical assay of GUS activity. Cold-activation of BN115 was strongly inhibited by the membrane fluidizer benzyl alcohol, but mimicked at 25 degrees C by the membrane rigidifier dimethylsulfoxide (DMSO). The cold induction of BN115 was also inhibited by stabilizers of microtubules and actin microfilaments, taxol and jasplakinolide, respectively, but was mimicked at 25 degrees C by microtubule destabilizer oryzalin or colchicine, or by microfilament destabilizer latrunculin B. Gd3+ or ruthenium red prevented the cold activation of BN115, but Ca2+ ionophore A23187 or cyclic ADP-ribose activated it at 25 degrees C. Inhibitors of tyrosine kinases, protein kinase C and phosphoinositide kinases prevented the cold activation of BN115, but inhibitors of protein phosphatases (PP) 1 and 2 A activated BN115 at 25 degrees C. Constitutively expressed GUS activity in another transgenic line of the same cultivar of B. napus, was not affected by cold or any of the chemical treatments used in the experimentation. Activation of BN115 at 25 degrees C by DMSO, Ca2+ ionophore, cADPR, and by inhibitors of PP1 and 2A was accompanied by an increased freezing tolerance. It was concluded that the cold-activation of BN115 requires membrane rigidification, cytoskeleton reorganization, Ca2+ influx and action of several types of protein kinases. PMID- 11489179 TI - In vitro interactions between barley TALE homeodomain proteins suggest a role for protein-protein associations in the regulation of Knox gene function. AB - This paper describes two-hybrid interactions amongst barley homeodomain proteins encoded by the Three Amino acid Loop Extension (TALE) superfamily. The class I KNOX protein BKN3 is shown to homodimerise and to associate with proteins encoded by the class I and II Knox genes BKn-1 and BKn-7. Furthermore, JUBEL1 and JUBEL2, two BELL1 homologous proteins, are identified and characterised as interacting partners of BKN3. Differences in the requirements of BKN3 derivatives for interactions with KNOX and JUBEL proteins imply the involvement of overlapping but slightly different domains. This set of results is an example for interactions amongst different classes of plant TALE homeodomain proteins, as previously described for related animal proteins. Apparently identical spatial and temporal expression patterns of BKn-1, BKn-3, BKn-7, JuBel1 and JuBel2, as determined by in situ hybridisation, are compatible with possible interactions of their protein products in planta. Contradictory to the common model, that the transcriptional down-regulation of certain class 1 Knox-genes is the prerequisite for organ differentiation, transcripts of all five genes were, similar to Tkn1 and Tkn2/LeT6 of tomato, detected in incipient and immature leaves as well as in meristematic tissues. A characteristic phenotype is induced by the overexpression of JuBel2 in transgenic tobacco plants. PMID- 11489180 TI - Hsp101 is necessary for heat tolerance but dispensable for development and germination in the absence of stress. AB - Hsp101 is a molecular chaperone that is required for the development of thermotolerance in plants and other organisms. We report that Arabidopsis thaliana Hsp101 is also regulated during seed development in the absence of stress, in a pattern similar to that seen for LEA proteins and small Hsps; protein accumulates during mid-maturation and is stored in the dry seed. Two new alleles of the locus encoding Hsp101 (HOT1) were isolated from Arabidopsis T-DNA mutant populations. One allele, hot1-3, contains an insertion within the second exon and is null for Hsp101 protein expression. Despite the complete absence of Hsp101 protein, plant growth and development, as well as seed germination, are normal, demonstrating that Hsp101 chaperone activity is not essential in the absence of stress. In thermotolerance assays hot1-3 shows a similar, though somewhat more severe, phenotype to the previously described missense allele hot1 1, revealing that the hot1-1 mutation is also close to null for protein activity. The second new mutant allele, hot1-2, has an insertion in the promoter 101 bp 5' to the putative TATA element. During heat stress the hot1-2 mutant produces normal levels of protein in hypocotyls and 10-day-old seedlings, and it is wild type for thermotolerance at these stages. Thus this mutation has not disrupted the minimal promoter sequence required for heat regulation of Hsp101. The hot1-2 mutant also expresses Hsp101 in seeds, but at a tenfold reduced level, resulting in reduced thermotolerance of germinating seeds and underscoring the importance of Hsp101 to seed stress tolerance. PMID- 11489181 TI - Localization of flavonoid enzymes in Arabidopsis roots. AB - Immunofluorescence and immuno-electron microscopy have been used to test the hypothesis that flavonoid metabolism is organized as a membrane-associated enzyme complex. The cellular and subcellular locations of chalcone synthase (CHS) and chalcone isomerase (CHI), the first two enzymes of this pathway, were examined in Arabidopsis roots. High levels of both enzymes were found in the epidermal and cortex cells of the elongation zone and the root tip, consistent with the accumulation of flavonoid endproducts at these sites. Co-localization of CHS and CHI was observed at the endoplasmic reticulum and tonoplast in these cells, and also in electron-dense regions that are, as yet, unidentified. In addition, a striking asymmetric distribution was observed for these enzymes in cortex cells of the elongation zone, which may provide clues about the physiological function of flavonoids in roots. The accumulation of CHS and CHI was also examined in tt7(88), a mutant in the gene for flavonoid 3'-hydroxylase (F3'H), which has been postulated to serve as a membrane anchor for the flavonoid enzyme complex. CHS and CHI accumulated to lower levels in cortex cells and higher levels in epidermal cells in the roots of this mutant as compared with wild-type plants. Moreover, the electron-dense regions containing these two enzymes were not observed. However, localization of CHS and CHI to the ER and tonoplast did not appear to be affected, suggesting that other proteins may function in recruiting the "soluble" flavonoid enzymes to membranes. Staining of flavonoid endproducts with DPBA was consistent with expression of CHS and CHI in these seedlings. PMID- 11489182 TI - Integration of the FISH pachytene and genetic maps of Medicago truncatula. AB - A molecular cytogenetic map of Medicago truncatula (2n = 2x = 16) was constructed on the basis of a pachytene DAPI karyogram. Chromosomes at this meiotic prophase stage are 20 times longer than at mitotic metaphase, and display a well differentiated pattern of brightly fluorescing heterochromatin segments. We describe here a pachytene karyogram in which all chromosomes can be identified based on chromosome length, centromere position, heterochromatin patterns, and the positions of three repetitive sequences (5S rDNA, 45S rDNA and the MtR1 tandem repeat), visualized by fluorescence in situ hybridization (FISH). We determined the correlation between genetic linkage groups and chromosomes by FISH mapping of bacterial artificial chromosome (BAC) clones, with two to five BACs per linkage group. In the cytogenetic map, chromosomes were numbered according to their corresponding linkage groups. We determined the relative positions of the 20 BACs and three repetitive sequences on the pachytene chromosomes, and compared the genetic and cytological distances between markers. The mapping resolution was determined in a euchromatic part of chromosome 5 by comparing the cytological distances between FISH signals of clones of a BAC contig with their corresponding physical distance, and showed that resolution in this region is about 60 kb. The establishment of this FISH pachytene karyotype, with a far better mapping resolution and detection sensitivity compared to those in the highly condensed mitotic metaphase complements, has created the basis for the integration of molecular, genetic and cytogenetic maps in M. truncatula. PMID- 11489183 TI - A method for isolating a high yield of Arabidopsis chloroplasts capable of efficient import of precursor proteins. AB - Chloroplasts were isolated from Arabidopsis plants grown under different conditions, and using different protocols, to determine a method that would yield chloroplasts capable of binding and importing precursor proteins. Chloroplasts isolated from protoplasts and purified on a Percoll gradient were highly import competent, with little non-specific binding of the precursor, and a high yield of intact chloroplasts (0.1 mg chlorophyll/g FW). Chloroplasts from plants grown on agar plates had a much higher rate of import than those from plants grown on soil. Protein import remained high at all of the ages tested for chloroplasts from plate-grown plants, whereas it declined during the development of soil-grown plants. Arabidopsis chloroplasts imported a range of precursor proteins and had nucleotide requirements for binding and import similar to those reported for pea chloroplasts. PMID- 11489184 TI - Quantitative in vivo measurement of glutathione in Arabidopsis cells. AB - A new, non-destructive assay is described to quantify cytoplasmic glutathione (GSH) levels in vivo in single cells or populations of cells from Arabidopsis suspension cultures. Cytoplasmic GSH was labelled with monochlorobimane (MCB) in situ to give a fluorescent GSH-bimane (GSB) conjugate. At low (10-100 microM) concentrations of MCB, labelling was mediated by a glutathione S-transferase, which confers specificity for GSH. HPLC analysis of MCB-labelled low molecular weight thiols showed that the assay measures the total GSH pool, including the oxidized glutathione. The progress curve for the labelling could be described using Michaelis-Menten kinetics with an apparent KM of 40 microM and Vmax of 470 micromol lcyt -1 min-1. There was no evidence for de novo synthesis of GSH during the labelling period of 2 h, suggesting that control of GSH synthesis is not mediated by feedback control of gamma-glutamylcysteine synthetase in this system. The total cellular level of GSH was calculated from the plateau value of the progress curve, after appropriate calibration, as 830-942 nmol g-1 FW. The volume fraction of cytoplasm was measured from serial optical sections of bimane labelled cells collected by confocal laser scanning microscopy (CLSM) with excitation 442 nm, or two-photon laser scanning microscopy (TPLSM) with excitation 770 nm. A value of 42 +/- 3% cytoplasm was determined by manual segmentation, and a value of 37 +/- 2% using stereological techniques. Using these figures, values for cytoplasmic [GSH] were estimated to be between 2.7 +/- 0.3 and 3.2 +/- 0.3 mM for cell populations. In addition, measurement of GSH levels in individual cells using CLSM and TPLSM gave values of 3.0 +/- 0.5 and 3.5 +/- 0.7 mM, respectively. PMID- 11489185 TI - Inducible high-level mRNA amplification system by viral replicase in transgenic plants. AB - We have constructed a new system for inducible high-level expression of mRNA for foreign genes in transgenic plants by introducing a glucocorticoid-inducible transcription system into the previously developed "mRNA amplification system" where target mRNA can be amplified as a subgenomic RNA by the replicase of a plant tripartite RNA virus, Brome mosaic virus (BMV). In the new amplification system, the amplification of mRNA is tightly regulated by the expression of a subunit of the BMV replicase. Transgenic Nicotiana benthamiana plants (designated GVG1 x 2FR) were produced that contained cDNA of BMV RNA1 coding a subunit of replicase under the control of a tightly regulated, glucocorticoid-inducible promoter. In addition GVG1 x 2FR plants contain cDNAs of BMV RNA2 coding another subunit of the replicase, and a replicable engineered BMV RNA3 derivative (FCP2IFN) carrying the human gamma interferon (IFN) gene under the control of the Cauliflower mosaic virus 35S promoter. When transgenic plants were treated with dexamethasone (DEX), a strong synthetic glucocorticoid, induction of replication and amplification of the 35S-driven FCP2IFN and synthesis of subgenomic mRNA for IFN were observed. Accumulation levels of amplified FCP2IFN were over 300 times higher than those of the 35S-driven FCP2IFN in the GVG1 x 2FR plant without the treatment and those of the mRNA for IFN were 30-230 times higher than in the previous, non-inducible mRNA amplification system. Without DEX treatment, no subgenomic mRNA for IFN was detected in the GVG1 x 2FR plant. The advantages and potential uses of this system are also discussed. PMID- 11489187 TI - A deficiency of coproporphyrinogen III oxidase causes lesion formation in Arabidopsis. AB - We isolated an Arabidopsis lesion initiation 2 (lin2) mutant, which develops lesion formation on leaves and siliques in a developmentally regulated and light dependent manner. The phenotype of the lin2 plants resulted from a single nuclear recessive mutation, and LIN2 was isolated by a T-DNA tagging approach. LIN2 encodes coproporphyrinogen III oxidase, a key enzyme in the biosynthetic pathway of chlorophyll and heme, a tetrapyrrole pathway, in Arabidopsis. The lin2 plants express cytological and molecular markers associated with the defense responses, usually activated by pathogen infection. These results demonstrate that a porphyrin pathway impairment is responsible for the lesion initiation phenotype, which leads to the activation of defense responses, in Arabidopsis. Lesion formation was not suppressed, and was even enhanced when accumulation of salicylic acid (SA) was prevented in lin2 plants by the expression of an SA degrading salicylate hydroxylase (nahG) gene. This suggests that the lesion formation triggered in lin2 plants is determined prior to or independently of the accumulation of SA but that the accumulation is required to limit the spread of lesions in lin2 plants. PMID- 11489188 TI - Evidence for a disease-resistance pathway in rice similar to the NPR1-mediated signaling pathway in Arabidopsis. AB - The Arabidopsis NPR1/NIM1 gene is a key regulator of systemic acquired resistance (SAR). Over-expression of NPR1 leads to enhanced resistance in Arabidopsis. To investigate the role of NPR1 in monocots, we over-expressed the Arabidopsis NPR1 in rice and challenged the transgenic plants with Xanthomonas oryzae pv. oryzae (Xoo), the rice bacterial blight pathogen. The transgenic plants displayed enhanced resistance to Xoo. RNA blot hybridization indicates that enhanced resistance requires expression of NPR1 mRNA above a threshold level in rice. To identify components mediating the resistance controlled by NPR1, we used NPR1 as bait in a yeast two-hybrid screen. We isolated four cDNA clones encoding rice NPR1 interactors (named rTGA2.1, rTGA2.2, rTGA2.3 and rLG2) belonging to the bZIP family. rTGA2.1, rTGA2.2 and rTGA2.3 share 75, 76 and 78% identity with Arabidopsis TGA2, respectively. In contrast, rLG2 shares highest identity (81%) to the maize liguleless (LG2) gene product, which is involved in establishing the leaf blade-sheath boundary. The interaction of NPR1 with the rice bZIP proteins in yeast was impaired by the npr1-1 and npr1-2 mutations, but not by the nim1-4 mutation. The NPR1-rTGA2.1 interaction was confirmed by an in vitro pull-down experiment. In gel mobility shift assays, rTGA2.1 binds to the rice RCH10 promoter and to a cis-element required sequence-specifically for salicylic acid responsiveness. This is the first demonstration that the Arabidopsis NPR1 gene can enhance disease resistance in a monocot plant. These results also suggest that monocot and dicot plants share a conserved signal transduction pathway controlling NPR1-mediated resistance. PMID- 11489189 TI - HSR203 antisense suppression in tobacco accelerates development of hypersensitive cell death. AB - Activation of the tobacco gene hsr203 is rapid, highly localized, specific for incompatible plant-pathogen interactions, and strongly correlated with programmed cell death occurring in response to diverse pathogens. Functional characterization of hsr203 gene product has shown that HSR203 is a serine hydrolase that displays esterase activity. We show here that transgenic tobacco plants deficient in HSR203 protein exhibit an accelerated hypersensitive response when inoculated with an avirulent strain of Ralstonia solanacearum. This response was accompanied by a maximal level of cell death and a drastic inhibition of in planta bacterial growth. Transgenic plants deficient in HSR203 were also found to show increased resistance in a dosage-dependent manner to Pseudomonas syringae pv. pisi, another avirulent bacterial pathogen, and to virulent and avirulent races of Phytophthora parasitica, a fungal pathogen of tobacco, but not to different virulent bacteria. Surprisingly, expression of another hsr gene, hsr515, and that of the defence genes PR1-a and PR5, was strongly reduced in the transgenic lines. Our results suggest that hsr203 antisense suppression in tobacco can have pleiotropic effects on HR cell death and defence mechanisms, and induces increased resistance to different pathogens. PMID- 11489190 TI - Two types of HKT transporters with different properties of Na+ and K+ transport in Oryza sativa. AB - It is thought that Na+ and K+ homeostasis is crucial for salt-tolerance in plants. To better understand the Na+ and K+ homeostasis in important crop rice (Oryza sativa L.), a cDNA homologous to the wheat HKT1 encoding K+-Na+ symporter was isolated from japonica rice, cv Nipponbare (Ni-OsHKT1). We also isolated two cDNAs homologous to Ni-OsHKT1 from salt-tolerant indica rice, cv Pokkali (Po OsHKT1, Po-OsHKT2). The predicted amino acid sequence of Ni-OsHKT1 shares 100% identity with Po-OsHKT1 and 91% identity with Po-OsHKT2, and they are 66-67% identical to wheat HKT1. Low-K+ conditions (less than 3 mM) induced the expression of all three OsHKT genes in roots, but mRNA accumulation was inhibited by the presence of 30 mM Na+. We further characterized the ion-transport properties of OsHKT1 and OsHKT2 using an expression system in the heterologous cells, yeast and Xenopus oocytes. OsHKT2 was capable of completely rescuing a K+ uptake deficiency mutation in yeast, whereas OsHKT1 was not under K+-limiting conditions. When OsHKTs were expressed in Na+-sensitive yeast, OsHKT1 rendered the cells more Na+-sensitive than did OsHKT2 in high NaCl conditions. The electrophysiological experiments for OsHKT1 expressed in Xenopus oocytes revealed that external Na+, but not K+, shifted the reversal potential toward depolarization. In contrast, for OsHKT2 either Na+ or K+ in the external solution shifted the reversal potential toward depolarization under the mixed Na+ and K+ containing solutions. These results suggest that two isoforms of HKT transporters, a Na+ transporter (OsHKT1) and a Na+- and K+-coupled transporter (OsHKT2), may act harmoniously in the salt tolerant indica rice. PMID- 11489191 TI - Generation and properties of ascorbic acid-deficient transgenic tobacco cells expressing antisense RNA for L-galactono-1,4-lactone dehydrogenase. AB - In higher plants, the terminal step of L-ascorbic acid (AsA) biosynthesis is catalyzed by the enzyme L-galactono-1,4-lactone dehydrogenase (EC 1.3.2.3, GalLDH). We generated AsA-deficient transgenic tobacco BY-2 cell lines by antisense expression of the GalLDH cDNA that was amplified from BY-2 cells using PCR. Two transgenic cell-lines, AS1-1 and AS2-2, having a marked expression of antisense RNA were analyzed. Antisense suppression of GalLDH mRNA led to a significant decline in the GalLDH activity. The AsA levels in the transgenic cell lines were found to be 30% lower than the wild-type BY-2 cells. In synchronous cultures, division of AS1-1 and AS2-2 cells was restrained with a concomitant decrease in mitotic index that was probably due to a decline in AsA levels. The rate of cell growth was also found to be less than that of the wild-type cells. Interestingly, there was a significant phenotypic difference between the transgenic and wild-type cells. The calli of AS1-1 and AS2-2 appeared to be sticky and soft. Back extrusion method also showed that AsA-deficient BY-2 callus was rheologically soft. Furthermore, microscopic analysis revealed that AS1-1 and AS2-2 cells were abnormally slender, suggesting a potential for a significant and a uni-axial elongation. Thus, we observed that decline in the AsA levels has an adverse effect on the division, growth and structure of a plant cell. PMID- 11489192 TI - Partial complementation of embryo defective mutations: a general strategy to elucidate gene function. AB - The EMB 506 gene has been characterised as essential for embryo development. To provide insights into the role of EMB 506, which is hidden by the embryo defective phenotype, the ABI3 promoter was fused to the EMB 506 cDNA. The expression of such a transgene should provide sufficient protein during embryogenesis to ensure normal embryo development in homozygous emb 506 seeds. We show that homozygous emb 506 seedlings, partially complemented with the ABI3::EMB 506 transgene, can be obtained. Most of the rescued emb 506 plants are able to flower and to set normal seeds, but show mild to severe depigmentation of rosette leaves and/or inflorescences. This effect on chloroplast development indicated a putative chloroplast localisation of the EMB 506 protein, which was demonstrated by GFP-protein fusion. However, EMB 506 cannot be considered as a chloroplast housekeeping protein only, since EMB 506 is not present in all photosynthetic tissues. This study demonstrates the power of this simple strategy, which could be widely applied to other emb mutants and which may reveal similar or additional roles for EMB genes at vegetative stages of the life cycle. PMID- 11489193 TI - Multiple pathways for Cre/lox-mediated recombination in plastids. AB - Plastid transformation technology involves the insertion by homologous recombination and subsequent amplification of plastid transgenes to approximately 10 000 genome copies per leaf cell. Selection of transformed genomes is achieved using a selectable antibiotic resistance marker that has no subsequent role in the transformed line. We report here a feasibility study in the model plant tobacco, to test the heterologous Cre/lox recombination system for antibiotic marker gene removal from plastids. To study its efficiency, a green fluorescent protein reporter gene activation assay was utilized that allowed visual observation of marker excision after delivery of Cre to plastids. Using a combination of in vivo fluorescence activation and molecular assays, we show that transgene excision occurs completely from all plastid genomes early in plant development. Selectable marker-free transplastomic plants are obtained in the first seed generation, indicating a potential application of the Cre/lox system in plastid transformation technology. In addition to the predicted transgene excision event, two alternative pathways of Cre-mediated recombination were also observed. In one alternative pathway, the presence of Cre in plastids stimulated homologous recombination between a 117 bp transgene expression element and its cognate sequence in the plastid genome. The other alternative pathway uncovered a plastid genome 'hot spot' of recombination composed of multiple direct repeats of a 5 bp sequence motif, which recombined with lox independent of sequence homology. Both recombination pathways result in plastid genome deletions. However, the resultant plastid mutations are silent, and their study provides the first insights into tRNA accumulation and trans-splicing events in higher plant plastids. PMID- 11489194 TI - Efficient elimination of selectable marker genes from the plastid genome by the CRE-lox site-specific recombination system. AB - Incorporation of a selectable marker gene during transformation is essential to obtain transformed plastids. However, once transformation is accomplished, having the marker gene becomes undesirable. Here we report on adapting the P1 bacteriophage CRE-lox site-specific recombination system for the elimination of marker genes from the plastid genome. The system was tested by the elimination of a negative selectable marker, codA, which is flanked by two directly oriented lox sites (>codA>). Highly efficient elimination of >codA> was triggered by introduction of a nuclear-encoded plastid-targeted CRE by Agrobacterium transformation or via pollen. Excision of >codA> in tissue culture cells was frequently accompanied by a large deletion of a plastid genome segment which includes the tRNA-ValUAC gene. However, the large deletions were absent when cre was introduced by pollination. Thus pollination is our preferred protocol for the introduction of cre. Removal of the >codA> coding region occurred at a dramatic speed, in striking contrast to the slow and gradual build-up of transgenic copies during plastid transformation. The nuclear cre gene could subsequently be removed by segregation in the seed progeny. The modified CRE-lox system described here will be a highly efficient tool to obtain marker-free transplastomic plants. PMID- 11489196 TI - Quantitation of dialysis: historical perspective. AB - This article is an attempt to provide a historical perspective to the ongoing attempts to quantify dialysis therapy. It is immediately apparent that motivated chemists, physicists, engineers, mathematicians, and other scientists from all over the world have greatly aided this effort. Dialysis, described by Graham in 1861, was furthered by Abel et al. and Hass before World War I. Willem Kolff attempted to evaluate mass removed and Alwall used a solute extraction ratio. However, the concept of "clearance" and "dialysance" awaited the studies of Wolf et al. in 1951. This classic work describes most of the information concerning actual dialyzer performance known today. A. S. Michaels provided the equations leading to the KoA/Ro/A concept in 1966 which only very recently required updating. The interaction of diffusion and convection is complex and was studied by Villarroel in 1977 and recently by Jaffrin. L. W. Henderson studied and described hemofiltration and hemodiafiltration from 1967-1975. Efforts to relate the patient's outcome to the dialyzer's performance have been difficult and ongoing since 1971; the Babb-Scribner Square meter-hour (which included the expression "Kt/V"); the Kopp et al. Liter-Kilogram concept; 1972 Kjellstrand clearance * time/kg or Liter. A NIH sponsored conference on the Adequacy of Dialysis in Monterey, California in March of 1974 was focused somewhat on the "middle molecule" theory of uremic toxicity, but contained a presentation by Sargent and Gotch on the possibilities of urea kinetic modeling. They developed iterative computer programs to obtain the best estimates of the required variables. At about this same time, Teschan, Ginn et al. published a series of neurofunctional tests and EEG power spectra analyses which most convincingly showed that dialysis two times a week was inadequate, and that dialysis delivered three times a week at urea clearance equal to body water volume was required to normalize these abnormalities: a major contribution! The National Cooperative Dialysis Study reported in Kidney International, 1983, was either misunderstood or ignored by most practitioners. The mechanistic analysis of the study by Gotch and Sargent appeared in 1985 and indicated that at adequate protein intake a Kt/V >0.8 yielded better patient survival. In 1982 Malchesky reported the Direct Dialysis Quantification (DDQ) based on calculations from the total mass removed in the dialysate. Although cumbersome, it avoids many errors including the effect of hematocrit and other factors on dialyzer clearance and many consider it to be "the gold standard." The 1990s were characterized by the development of many simple logarithmic equations to estimate Kt/V and eKt/V suitable for spreadsheets which could be used for CQI by individual units. These are primarily by J. T. Daugirdas and coworkers, Smye and Tattersall. In 1991 the Urea Reduction Ratio (URR) was introduced by Lowrie, who in 1999 suggested that Kt and V (as indicator of lean body mass) were independent predictors of survival. Peritoneal dialysis: Although performed before and immediately after World War II, almost all of the basic quantification mechanistics and data are found in the publications of S. T. Boen (1964). New quantifiers, the Mass Transport Area Coefficient (MTAC) or Pyle Popovich model, the Henderson-Nolph, and Garred models, were compared by Waniewski. Gotch announced a PD modeling program which suggested that a weekly PKt/V at 2.1 was needed to supply the same urea removal as a Kt/V of 3.6, but warned that both were sensitive to decreased time. PMID- 11489198 TI - Evolution of the single-pool urea kinetic model. AB - Our interest in urea kinetic modeling (UKM) was stimulated some 30 years ago at the time of the advent of hollow fiber kidneys with greatly improved urea transport. This led to examination of the interaction between time and clearance in computing the dialysis dose. In early studies a fixed-volume single-pool UKM was used but this frequently gave spurious high volumes and led to the advent of the variable-volume single-pool model. The role of volume calculation in assessment of the delivered dialysis dose and the value of normalized protein catabolic rate (nPCR) calculation are reviewed. More recently quantification of double-pool effects has become simplified and now is widely used for UKM. The National Cooperative Dialysis Study (NCDS) resulted in the concept of dose quantification by Kt/V. This is reviewed, including the controversy surrounding interpretation of the NCDS. Currently there is great interest in more frequent dialysis, 4-6 days/week. The development of a new dose parameter, the standard Kt/V (stdKt/V), to enable quantitative comparison of dose with widely varying dose schedules is discussed. PMID- 11489197 TI - Uremic toxicity: urea and beyond. AB - Successful replacement of renal function with dialysis supports the concept that uremia is a toxic state resulting from accumulated solutes and that toxicity results from high concentrations of these solutes in body fluids. Dialyzer clearance of urea, a surrogate toxin, is the currently accepted best measure of dialysis and dialysis adequacy, but it is admittedly a compromise due to current lack of knowledge about and inability to measure more toxic solutes. This failure could be explained if uremic toxicity is actually a summation effect of multiple toxins, each at individual subtoxic levels in the patient. Other solutes could be used as surrogates to measure clearance, but urea happens to be available in high concentrations, is easily measured by all clinical laboratories, and is easily dialyzed, so changes in concentration are sensitive indicators of clearance. Measurements of creatinine clearance are confounded by the disequilibrium that occurs across red cells within the dialyzer and in the patient. Other solutes probably behave more like creatinine than urea, so urea stands out as uniquely diffusible, a property that actually spoils its effectiveness as a surrogate toxin, especially when applied to more frequent and continuous dialysis. Accumulation of other solutes may correlate better with toxic uremic symptoms and the residual syndrome. More studies are needed to examine the kinetics of other solutes, their generation rates, and their distribution volumes to provide clinicians with more knowledge and tools to optimize dialysis treatments. Examination of the effectiveness of solute removal in patients dialyzed more frequently may provide significant insight into the pathogenesis of uremia. PMID- 11489199 TI - Factors affecting hemodialysis and peritoneal dialysis efficiency. AB - Hemodialysis and peritoneal dialysis are two blood purification techniques that use similar operating systems. The hemodialysis system is based on three components (blood, membrane, and dialysate). The peritoneal dialysis system is based on the same components that can, however, be less manipulated and adjusted. In hemodialysis the blood flow is the main determinant of small solute removal thanks to a prevalently diffusive mechanism. Convection is also used to transport larger solutes across the membrane, but this mechanism relies on the high permeability coefficient of the membrane and high transmembrane pressure leading to high ultrafiltration rates. The membrane can therefore influence the performance of the techniques as far as solute removal and ultrafiltration are concerned. Finally, diffusion is facilitated by an improved distribution of dialysate flow in the dialysate compartment. This can be achieved with a special dialysate pathway configuration based on space yarns or micronodulation of the fibers. In peritoneal dialysis, blood flow and membrane characteristics can be less manipulated or almost not at all. The only variables are dialysate volume, flow, dwell time, and composition. Thanks to modification in these aspects of the dialysate, peritoneal dialysis techniques with different clearances and ultrafiltration rates can be accomplished. PMID- 11489200 TI - Past, present, and future of quantified peritoneal dialysis. AB - The role of peritoneal dialysis (PD) as a modality in renal replacement therapy has been well established. In this article we review various aspects in the evolution of PD, with special emphasis on adequacy. Until the late 1950s PD was still considered as a last resort in the treatment of terminal uremia. The introduction of a chronic indwelling catheter made chronic PD possible. The concept of continuous ambulatory peritoneal dialysis (CAPD), proposed in 1975, had a major impact on the way PD was performed later. The value of determining the adequacy, using urea clearance normalized to total body water (Kt/V) or creatinine clearance normalized to body surface area, was clearly highlighted by the Canada-USA (CANUSA) study. Introduction of standardized peritoneal equilibration tests has been very helpful in enhancing the efficiency of PD. In 1995 the National Kidney Foundation-Dialysis Outcomes Quality Initiatives (NKF DOQI) established guidelines to improve patient survival and outcome on dialysis. These guidelines established minimum criteria for PD adequacy. Compliance and malnutrition remain important factors determining the efficacy of PD. The "healthy start" concept emphasizes an early start of dialysis in patients with end-stage renal disease (ESRD). The quest for an ideal PD modality has recently led to renewed interest in the idea of continuous flow peritoneal dialysis (CFPD). PD continues to grow and at the same time faces many challenges. Its role as a renal replacement therapy is likely to evolve further in the years to come. PMID- 11489201 TI - Explaining counter-intuitive clinical outcomes predicted by Kt/V. AB - Population-based studies of maintenance hemodialysis patients have demonstrated a reproducible relationship between the dose of hemodialysis and mortality and morbidity outcomes. In these analyses, which have aggregated hemodialysis patient subgroups, improved outcomes are associated with greater doses of hemodialysis. However, remarkable counterintuitive findings are observed if patients are analyzed by subgroups based on their race, gender, and anthropometric and blood based biomarkers of nutritional state. For example, blacks generally receive lower doses of hemodialysis than whites, but enjoy relatively improved survival; patients who receive the highest doses of hemodialysis have an increased death risk; and the dose response curve between hemodialysis and survival is altered based on the patients' body mass index. These seemingly paradoxical relationships between hemodialysis dose and patient survival can be explained because of the use of mathematical urea kinetic constructs as clinical outcome predictors; they integrate a measure of solute removal (K x t) with an anthropometric surrogate of nutrition, the urea distribution volume (V). Both these measures have an independent influence on patient survival and in some clinical circumstances are of unequal power as clinical outcome predictors. These complex interactions must be kept in perspective as clinical care is delivered in the context of hemodialysis dose. PMID- 11489202 TI - Compartment effects in hemodialysis. AB - Compartment effects in hemodialysis are important because they reduce the efficiency of removal of the compartmentalized solute during dialysis. The dialyzer can only remove those waste products that are presented to it, and then only in proportion to the concentration of the solute in the blood. Classically a two-compartment system has been modeled, with the compartments arranged in series. Because modeling suggests that the sequestered compartment is larger than the accessible compartment, an assumption has been made that the sequestered compartment is the intracellular space. For urea and other solutes that move easily across many cell membranes, compartmentalization may be flow related, that is, related to sequestration in organs (muscle, skin, bone). Although mathematically urea rebound and mass balance can be described with either model, the flow-related model best explains data showing that urea rebound after dialysis is increased during ultrafiltration, diminished during high cardiac output states, and also reduced during exercise. Whether compartmentalization is increased in vasoconstricted intensive care unit patients receiving acute dialysis remains an open question. PMID- 11489203 TI - Significance of distribution volume in dialysis quantification. AB - The urea distribution volume is in several ways an important parameter in the treatment of end-stage renal disease (ESRD) patients. It has a major impact on the relative dose of dialysis treatment as measured by Kt/V. It is also important in the assessment of fluid status, which has a direct influence on blood pressure. Nevertheless, urea distribution volume is usually not measured on a regular basis, probably because it has been perceived as a relatively complicated measurement. With the arrival of on-line monitors for dialysate urea this situation has improved radically. In this article a number of volume determination methods are discussed, including some new methods based on on-line dialysate urea monitoring, which are shown to perform well in comparison to reference methods. PMID- 11489204 TI - Effect of therapy time and frequency on effective solute removal. AB - Increasing dialysis frequency or time increases the removal of the molecules diffusing slowly across the intercompartmental barriers. By offering frequent dialysis the time on dialysis can be decreased, possibly without worsening the outcome. Increasing dialysis time increases large molecule removal. Increasing in both frequency and time on dialysis increases the removal of solutes of all molecular sizes. Increasing frequency and/or time of dialysis may have many other beneficial effects that are not traditionally quantitated and which can affect outcomes. PMID- 11489205 TI - Reconciling differences in effective solute removal between intermittent and continuous therapies. AB - Solute removal by various forms of renal replacement therapy (RRT) differs from that occurring in the native kidney in several ways. Among the dialytic therapies, the relationship between clearance and mass removal rate may differ substantially. The purpose of this article is to review the various approaches that have been proposed to account for this differing relationship among the various types of RRT. Specific quantitative approaches along with clinical applications are provided. PMID- 11489206 TI - Quantification of middle molecular weight solute removal in dialysis. AB - The pioneering work of Gotch emphasized the critical need to be quantitative with respect to treatment prescription. Through his meticulous derivations and analyses regarding Kt/V(urea), he has provided powerful insight into the standard therapy prescriptions that we now employ clinically. However, time has seen the proliferation of treatment techniques, most of which are too "young" to have been characterized with respect to clinical outcomes. Further, the relationship between removal of urea and removal of middle molecular size solutes associated with these newer techniques deviates from that associated with conventional, clinically qualified techniques. In this article we examine the solute clearance profile of some of these new methodologies and their relationship to current criteria for treatment adequacy. Our approach is to discuss components of the overall transport process and then utilize modeling of surrogate molecules over the size range of interest whose transport characteristics are known. Alteration in the solute clearance profile of these surrogate markers in response to changes in prescription variables will thus offer insight into the spectrum of toxic middle molecules that are removed when size, space of distribution, and generation rate are known. PMID- 11489207 TI - Future directions in dialysis quantification. AB - The influence of dialysis prescription on outcome is well established, and currently the amount of dialysis prescribed is based on small molecular weight toxin removal as represented by the clearance of urea. The "normalized dose of dialysis" (Kt/V(urea)) concept is well established. Most techniques for dialysis quantification require that blood samples be taken at the beginning and after the completion of dialysis. The postdialysis sample, however, gives cause for concern because of the "rebound phenomenon" due to nonuniform distribution of urea among body compartments. Blood samples give "indirect" measures of dialysis quantification. Thus direct urea concentration measurements in dialysate may be superior in urea kinetic modeling and these may be made "real time" during dialysis. It is with real-time monitoring that future advances in dialysis quantification will take place. These will be of two types. The first will analyze blood water or dialysate samples for urea content multiple times throughout the treatment; the second will assess the on-line clearance of urea using surrogate molecules such as sodium chloride, the clearance being determined by conductivity measurements. On-line urea monitoring is based on the action of urease on urea in a water solution and measurement of the resultant ammonium ions, which are measured directly by a specific electrode or indirectly by conductivity changes. Differences in blood-side versus dialysate-side urea monitors exist which reflect the parameters they can provide, but with both, the standard urea kinetic measurements of Kt/V and nPCR (nPNA) are easily obtainable. A range of additional parameters can be derived from dialysate-side monitoring such as "whole-body Kt/V," "pretreatment urea mass" and "whole-body urea clearance," which are worthy of future studies to determine their roles in adequacy assessment. Conductivity clearance measurements are made by examining the conductivity differences between dialysate inlet and outlet measured at two different dialysate inlet concentrations. This allows for the calculation of the electrolyte (ionic) dialysance, which is equal to the "effective" urea clearance, that is, the clearance that takes into account recirculation effects that reduce hemodialysis efficiency. The continuous reading of effective ionic clearance will allow an average value for K to be obtained for that dialysis, and hence the parameter K x t as an indication of dialysis dose is easily and accurately obtained for every treatment. The conductivity technology is cheap and rugged, and thus expanded use can be expected. Urea monitors have an inherent cost and require maintenance, and perhaps will remain researchers' tools for the present. The methodologies can complement each other; the addition of an accurate and independent value for K to dialysate based urea monitoring is like having simultaneous blood- and dialysate-side monitoring, and allows further increase in measurable parameters. PMID- 11489208 TI - I had a glassen vessel. PMID- 11489209 TI - The unfolded protein response: no longer just a special teams player. AB - The endoplasmic reticulum stress pathway known as the unfolded protein response is currently the best understood model of interorganellar signal transduction. Bridging a physical separation, the pathway provides a direct line of communication between the endoplasmic reticulum lumen and the nucleus. With the unfolded protein response, the cell has the means to monitor and respond to the changing needs of the endoplasmic reticulum. Beginning with the discovery of its remarkable signaling mechanism in yeast, the unfolded protein response has not ceased to reveal more of its many secrets. By applying powerful biochemical, genetic, genomic, and cytological approaches, the recent efforts of many groups have buried the long-held notion that the unfolded protein response is simply a regulatory platform for endoplasmic reticulum chaperones. We now know that the unfolded protein response regulates many genes that affect diverse aspects of cellular physiology. In addition, studies in mammals have revealed novel unfolded protein response signaling factors that may contribute to the specialized needs of multicellular organisms. This article focuses on these and other recent developments in the field. PMID- 11489210 TI - Approaching the molecular mechanism of autophagy. AB - Autophagy is a complex cellular process that involves dynamic membrane rearrangements under a range of physiological conditions. It is a highly regulated process that plays a role in cellular maintenance and development, and has been implicated in a number of genetic diseases. Upon induction of autophagy, cytoplasm is sequestered into vesicles and delivered to a degradative organelle, the vacuole in yeast or the lysosome in mammalian cells. The process is unique in that it converts material that is topologically intracellular into topologically extracellular. Autophagy was first described more than 50 years ago, but it is since the discovery of the pathway in yeast cells that our knowledge about the molecular events taking place during the process has expanded. The generation of autophagy-specific mutants in a variety of yeast cell lines has provided insight into functional roles of more than 15 novel genes, double that number if we include genes whose products function also in other processes. Although we have learned much about autophagy, many questions remain to be answered. This review highlights the most recent advances in the autophagy field in both yeast and mammalian cells. PMID- 11489211 TI - Organization of the Rab-GDI/CHM superfamily: the functional basis for choroideremia disease. AB - Choroideremia is an X-chromosome-linked disease that leads to the degeneration of the choriocapillaris, the retinal pigment epithelium and the photoreceptor layer in the eye. The gene product defective in choroideremia, CHM, is identical to Rab escort protein 1 (REP1). CHM/REP1 is an essential component of the catalytic geranylgeranyltransferase II complex (GGTrII) that delivers newly synthesized small GTPases belonging to the RAB gene family to the catalytic complex for post translational modification. CHM/REP family members are evolutionarily related to members of the guanine nucleotide dissociation inhibitor (GDI) family, proteins involved in the recycling of Rab proteins required for vesicular membrane trafficking through the exocytic and endocytic pathways, forming the GDI/CHM superfamily. Biochemical and structural analyses have now revealed a striking parallel in the organization and function of these two families allowing us to generate a general model for GDI/CHM superfamily function in health and disease. PMID- 11489212 TI - Qualitative highly divergent nuclear export signals can regulate export by the competition for transport cofactors in vivo. AB - Nucleo-cytoplasmic transport of proteins is mediated by nuclear export signals, identified in various proteins executing heterologous biological functions. However, the molecular mechanism underlying the orchestration of export is only poorly understood. Using microinjection of defined recombinant export substrates, we now demonstrate that leucine-rich nuclear export signals varied dramatically in determining the kinetics of export in vivo. Thus, nuclear export signals could be kinetically classified which correlated with their affinities for CRM1 containing export complexes in vitro. Strikingly, cotransfection experiments revealed that proteins containing a fast nuclear export signal inhibited export and the biological activity of proteins harboring a slower nuclear export signal in vivo. The affinity for export complexes seems therefore predominantly controlled by the nuclear export signal itself, even in the context of the complete protein in vivo. Overexpression of FG-rich repeats of nucleoporins affected a medium nuclear export signal containing protein to the same extent as a fast nuclear export signal containing protein, indicating that nucleoporins appear not to contribute significantly to nuclear export signal-specific export regulation. Our results imply a novel mode for controlling the biological activity of shuttle proteins already by the composition of the nuclear export signal itself. PMID- 11489213 TI - Selective control of basolateral membrane protein polarity by cdc42. AB - The rho GTPase cdc42 is implicated in several aspects of cell polarity. A recent study (Kroschewski R, Hall A, Mellman I. Nat Cell Biol 1999;1:8-13) demonstrated that a dominant negative mutant of cdc42 abolishes the polarity of basolateral membrane proteins in MDCK cells, but did not elucidate whether this effect was selective for basolateral proteins or nonselective for all secreted proteins. To answer this question, we analyzed the polarity of newly synthesized membrane and soluble proteins in MDCK cell lines previously induced to overexpress mutant forms of cdc42. GTPase-deficient and dominant negative cdc42 did not affect the apical targeting of a newly synthesized apical membrane protein, but reversed to apical the distribution of two exogenous basolateral membrane proteins. In striking contrast, GTPase-deficient cdc42 did not affect polarized exocytosis of endogenous soluble proteins, either apical or basolateral. The exquisitely selective regulation of polarized protein targeting by cdc42 may allow cells to fine-tune their membrane composition in response to extracellular signals during development, migration and in response to injury. PMID- 11489214 TI - Clathrin interactions with C-terminal regions of the yeast AP-1 beta and gamma subunits are important for AP-1 association with clathrin coats. AB - Heterotetrameric adaptor (AP) complexes are thought to coordinate cargo recruitment and clathrin assembly during clathrin-coated vesicle biogenesis. We have identified, and characterized the physiological significance of clathrin binding activities in the two large subunits of the AP-1 complex in Saccharomyces cerevisiae. Using GST-fusion chromatography, two clathrin-binding sites were defined in the beta1 subunit that match consensus clathrin-binding sequences in other mammalian and yeast clathrin-binding proteins. Clathrin interactions were also identified with the C-terminal region of the gamma subunit. When introduced into chromosomal genes, point mutations in the beta1 clathrin-binding motifs, or deletion of the gamma C-terminal region, reduced association of AP-1 with clathrin in coimmunoprecipitation assays. The beta1 mutations or the gamma truncation individually produced minor effects on AP-1 distribution by subcellular fractionation. However, when beta1 and gamma mutations were combined, severe defects were observed in AP-1 association with membranes and incorporation into clathrin-coated vesicles. The combination of subunit mutations accentuated growth and alpha-factor pheromone maturation defects in chc1-ts cells, though not to the extent caused by complete loss of AP-1 activity. Our results suggest that both the beta1 and gamma subunits contribute interactions with clathrin that are important for stable assembly of AP-1 complexes into clathrin coats in vivo. PMID- 11489215 TI - A Glut4-vesicle marker protein, insulin-responsive aminopeptidase, is localized in a novel vesicular compartment in PC12 cells. AB - Glut4-containing vesicles represent a regulated recycling compartment in insulin sensitive fat and skeletal muscle cells, the nature and origin of which are not fully understood. In addition to Glut4 itself, these vesicles compartmentalize a number of proteins, at least one of which, insulin-responsive aminopeptidase, or IRAP, is completely colocalized with Glut4 in insulin-sensitive tissues. However, unlike Glut4, IRAP is expressed in a variety of other tissues and cell lines. Here, we explored the intracellular localization of IRAP in the rat pheochromocytoma cell line PC12. We found that this protein is present in a distinct population of slowly recycling light vesicles. By gradient centrifugations, immunoadsorption and double immunofluorescent staining, these vesicles are different from transferrin-containing endosomes, small synaptic vesicles and secretory granules and may thus represent a novel compartment in PC12 cells. Glut4-GFP chimera transiently expressed in PC12 cells is targeted to IRAP-containing vesicles indicating that cotargeting of Glut4 and IRAP is not specific for adipocytes and myocytes, but is faithful in a foreign cell type. We suggest that PC12 cells may possess a novel type of a vesicular carrier that may represent the homolog of Glut4-vesicles. PMID- 11489216 TI - Measurement of venous blood flow in the human fetus: a dream comes true, but now for some standardization. PMID- 11489217 TI - Nuchal translucency in the first trimester of pregnancy: ten years on and still a pain in the neck? PMID- 11489218 TI - Outcome of pregnancy in chromosomally normal fetuses with increased nuchal translucency in the first trimester. AB - OBJECTIVES: To study the outcome of chromosomally normal pregnancies with increased nuchal translucency at the 10-14-week scan. DESIGN: Retrospective study of 1320 chromosomally normal singleton pregnancies with nuchal translucency of > or = 3.5 mm. In addition to fetal karyotyping these patients were managed with follow-up scans at 14-16 and 20-22 weeks, specialist fetal echocardiography and in selected cases by infection screening and further genetic testing. RESULTS: In the 1320 pregnancies there were 68 (5.15%) spontaneous abortions or intrauterine deaths, 18 (1.36%) neonatal and infant deaths and 154 (11.67%) terminations of pregnancy. In the 1080 (81.82%) survivors, 60 (5.56%) had abnormalities requiring medical or surgical treatment or leading to mental handicap. The chance of a livebirth with no defects in the group with nuchal translucency of 3.5-4.4 mm was 86%, for those with translucency of 4.5-5.4 mm it was 77%, for those with translucency of 5.5-6.4 mm it was 67%, and for those with translucency of > or = 6.5 mm it was 31%. CONCLUSIONS: Increased fetal nuchal translucency is associated with chromosomal abnormalities, many fetal defects and genetic syndromes. In the majority of cases a series of antenatal investigations, including fetal karyotyping, detailed scans, fetal echocardiography, as well as genetic testing and infection screening, that can be completed by 20 weeks of gestation would distinguish between the pregnancies destined to result in adverse outcome and those leading to the delivery of infants without major defects. PMID- 11489219 TI - Fetal nuchal translucency and normal chromosomes: a long-term follow-up study. AB - OBJECTIVE: To assess the impact of increased nuchal translucency observed during early pregnancy on the subsequent health of children with normal chromosomes, with special attention to cardiac anatomy and function. METHODS: Clinical examination and detailed cardiac evaluation were carried out in 50 chromosomally normal children at the age of 2.4-7.1 years who had had a nuchal translucency measurement of > or = 3 mm at 13-15 weeks' gestation. The data of two babies who died of heart defects were also included. RESULTS: Major cardiac defects were identified in four (8%) of the children. The growth of all children was within normal limits. One child had Noonan syndrome, one had a cleidocranial dysplasia and a third had a developmental delay together with an unrecognized syndrome. Webs in the neck region were noticed in two children, with no associated pathology. CONCLUSIONS: In chromosomally normal fetuses with increased nuchal translucency, fetal echocardiography is necessary to identify major cardiac defects. In general, the parents can be reassured that, in the great majority, postnatal development is normal. PMID- 11489220 TI - Sequential screening for trisomy 21 by nuchal translucency measurement in the first trimester and maternal serum biochemistry in the second trimester in a low risk population. AB - OBJECTIVE: To evaluate screening for trisomy 21 in a low-risk population utilizing a combination of nuchal translucency measurement in the first trimester and the triple test in the second trimester. METHODS: This was a retrospective study of 9342 women with singleton pregnancies who booked for delivery in our hospital over a period of 5 years. A nuchal translucency scan was carried out at 10-13 weeks' gestation and for those with a measurement of 3.5 mm or more chorionic villus sampling was performed. All other women were asked to return for the triple test at 16 weeks' gestation. Amniocentesis was offered to women in whom the nuchal translucency was 2.5-3.4 mm, the triple test showed a risk of > or = 1 : 250 and in women aged > or = 35 years. RESULTS: The detection rate using the combined screening method was 95% (18/19) with a screen-positive rate of 7.2%. In comparison, screening by maternal age alone would have identified nine (47%) trisomy 21 pregnancies with a screen-positive rate of 10.7%. CONCLUSION: Our data suggest that the combination of nuchal translucency measurement in the first trimester and the triple test in the second trimester is associated with a very high detection rate of trisomy 21 at a relatively low screen-positive rate. PMID- 11489221 TI - Screening for Down syndrome using first-trimester ultrasound and second-trimester maternal serum markers in a low-risk population: a prospective longitudinal study. AB - OBJECTIVES: To compare nuchal translucency and second-trimester maternal serum measurements as alternative methods of antenatal screening for Down syndrome in a low-risk population and to evaluate the consequence of combining the results in the estimation of risk. DESIGN: In a consecutive series of 4130 women aged less than 38 years with a singleton pregnancy, we examined both the detection rate of Down syndrome by nuchal translucency measurement at 10-14 weeks and maternal serum screening by human chorionic gonadotrophin and alpha-fetoprotein at 14-18 weeks. Women with a nuchal translucency measurement of > or = 3 mm and women with a maternal serum screening-derived risk > or = 1/250 were recommended to have amniocentesis. A second-trimester detailed ultrasound scan was also performed in all women. The outcome of all pregnancies was recorded prospectively and the detection rate and false-positive rate of different screening strategies were retrospectively analyzed. RESULTS: Out of the 4130 pregnancies that were followed (mean maternal age, 30.1 years), 12 cases of Down syndrome were observed (0.28%), all detected prenatally. Seven of 12 cases had a nuchal translucency measurement of > or = 3 mm (58%), and six out of 10 cases with available maternal serum screening had a calculated risk of > or = 1/250 (60%). Four of the five Down syndrome cases with a nuchal translucency measurement of < 3 mm were detected by subsequent maternal serum screening. At a threshold giving 5% of positive tests, the sensitivity of nuchal translucency, maternal serum screening and combined risk screening were 75%, 60% and 90%, respectively. CONCLUSIONS: In screening for Down syndrome, an approach which combines the results from first-trimester nuchal translucency and second-trimester biochemistry is effective and increases the detection rate compared to the use of any single test. However, this strategy is likely to raise the false-positive rate and the interpretation of maternal serum screening-derived risk should be combined with the first-trimester nuchal translucency measurement. PMID- 11489222 TI - Significance of chromosome 22q11 analysis after detection of an increased first trimester nuchal translucency. AB - OBJECTIVE: To determine the value of performing routine fluorescent in situ hybridization (FISH) for microdeletions of chromosome 22q11 when karyotyping fetuses with increased nuchal translucency. DESIGN: This was a prospective observational study carried out over an 18-month period. Fetal karyotyping by chorionic villus sampling was offered to 5429 women attending for a routine booking scan in the first trimester when their nuchal translucency adjusted risk for Down syndrome was > or = 1 in 300. Cytogenetic samples were routinely tested for the 22q11 microdeletion when the nuchal translucency was > or = 3 mm. RESULTS: The prevalence of increased nuchal translucency > or = 2.5 mm was 180 (3.3%) and > or = 3.5 mm was 42 (0.8%). None of 75 fetuses with an increased nuchal translucency and normal karyotype demonstrated a 22q11 microdeletion on FISH analysis. In the same cohort, 3 of 20 (15%) cases of major congenital heart defects in which nuchal translucency was measured, had a nuchal translucency measurement > or = 2.5 mm. CONCLUSIONS: Routine FISH analysis for chromosome 22q11 microdeletions at the time of chorionic villus sampling for increased first trimester nuchal translucency is of limited value. As a significant proportion of fetuses with increased nuchal translucency will be found to have congenital heart defects later in the pregnancy, FISH analysis for chromosome 22q11 microdeletions can be targeted to fetuses with specific congenital heart defects. Tissue from the chorionic villus sampling should therefore be stored for subsequent analysis, until after detailed echocardiography is performed. PMID- 11489223 TI - A comparative study of multifetal pregnancy reduction from triplets to twins in the first versus early second trimesters after detailed fetal screening. AB - OBJECTIVES: To compare the outcome of multifetal pregnancy reduction from triplets to twins performed either early (at 11-12 weeks' gestation) or late (at 13-14 weeks). METHODS: Ninety-five high-order pregnancies following assisted conception were studied. Transabdominal sonographically guided multifetal pregnancy reduction was performed early in 46 women, while 49 women first underwent a sonographic fetal anomaly scan before undergoing selective reduction. RESULTS: Sonographic screening led to selective termination of a specific fetus in nine cases due to increased nuchal translucency and relative intrauterine growth restriction in three cases each, and meningomyelocele, abdominal cyst and cystic hygroma in one case each. In the early reduction group a diagnosis of hypoplastic left heart in the two remaining twins was subsequently made, and one pair of twins suffers from cerebral palsy. The rate of pregnancy loss was not statistically different between the early (4.3%; 2/46) and late (4.0%; 2/49) termination groups. The birth weight and gestational age at birth were not statistically different between the early ( n = 85) and late ( n = 94) groups (2110 +/- 580 vs. 2140 +/- 490 g, and 35.8 +/- 3.0 vs. 35.7 +/- 3.5 weeks). Similarly there was no statistically significant difference between early and late groups in the incidence of very premature (24-32 weeks; 9.3 vs. 8.3%) and premature (33-36 weeks; 46.5 vs. 47.9%) births. CONCLUSIONS: Early second trimester multifetal pregnancy reduction from triplets to twins may allow more selective termination of abnormal fetuses without an adverse effect on the outcome of pregnancy. However, further studies are needed in order to confirm our observations in a larger series. PMID- 11489224 TI - Prenatal diagnosis of ventriculocoronary fistula. AB - BACKGROUND: Cardiac anomalies may be associated with abnormal coronary vascular connections. We report the prenatal diagnosis of ventriculocoronary fistula in three fetuses with associated cardiac anomalies. MATERIALS AND METHODS: Fetal echocardiography was performed in three patients referred for suspected cardiac anomaly. Two-dimensional fetal echocardiography was complemented by color Doppler flow imaging and spectral Doppler in all cases. RESULTS: A ventriculocoronary fistula was diagnosed in three patients referred at 22, 23 and 32 weeks. The first patient had hypoplastic left heart associated with transposition of the great arteries and pulmonary atresia with an intact interventricular septum. The coronary fistula arose from the transposed aorta to the left ventricle. In two patients ventriculocoronary fistula was found in association with pulmonary atresia and an intact interventricular septum. In all cases there was bidirectional flow within the fistula (diastolic blood flow towards the ventricle with reversal during ventricular systole). The pregnancy with hypoplastic left heart with transposition, and one of those with pulmonary atresia resulted in neonatal death and stillbirth, respectively. In the third instance the ventriculocoronary fistula was verified by postpartum cardiac angiography. The infant initially received a Blalock-Taussig shunt, subsequently replaced by a bidirectional Glenn shunt, and was doing well at the time of writing. CONCLUSION: A ventriculocoronary fistula can be identified prenatally by color and spectral Doppler. This anomaly should be sought in fetuses with outflow tract obstructive cardiac lesions and an intact interventricular septum. Prenatal diagnosis allows early angiography postnatally. Delineation of coronary vascular regions may therefore facilitate preoperative planning. PMID- 11489225 TI - Changes in fetal hemodynamics with ritodrine tocolysis. AB - OBJECTIVE: To evaluate the effect of ritodrine on the fetal cardiovascular system. METHODS: Cardiac and extracardiac Doppler waveforms were recorded in 12 fetuses prior to and during ritodrine therapy used for preterm labor. Maternal and fetal heart rates, the Doppler pulsatility indices of the umbilical artery, middle cerebral artery, descending thoracic aorta and renal artery, and time velocity integrals of the atrioventricular valves and the ductus arteriosus, were measured. RESULTS: Ritodrine infusion caused an increase in maternal and fetal heart rates, the left cardiac output as measured by the product of time velocity integral and heart rate, and the pulsatility index of the middle cerebral artery, and a decrease in the pulsatility index of the umbilical artery. CONCLUSIONS: Ritodrine infusion may alter placental and cerebral blood flow and may have a selective effect on the left side of the heart. PMID- 11489226 TI - Ventriculovascular physiology of the growth-restricted fetus. AB - OBJECTIVES: To examine the mechanisms by which intrauterine growth restriction may influence later cardiovascular risk by comparing the ventriculovascular physiology of gestational age- and weight-matched growth-restricted and normal fetuses. DESIGN: A prospective longitudinal observational study of 20 normal fetuses studied from 20 weeks to term at monthly intervals was compared with a growth-restricted cohort examined in the interval between diagnosis and delivery. The last values before delivery of the growth-restricted cohort were compared with the normal cohort in two analyses matched for weight and for gestation. Arterial and venous vessel wall physiology and aortic pulse wave velocity were examined longitudinally in the thoracic descending aorta and inferior vena cava using an ultrasonic phase-locked echo-tracking system. Serial echocardiographic examinations were performed assessing structure, ventricular dimensions, function and Doppler flows. RESULTS: There was a linear increase in cardiac preload and relative pulse amplitude in the inferior vena cava with gestation. In normal fetuses, the aortic pulse wave velocity, maximum incremental and late decremental velocities increased with gestation whilst the relative pulse amplitude decreased reflecting falling distal impedance. In both age- and weight-matched analyses, the growth-restricted fetuses showed significantly reduced values reflecting the chronic fetal ventriculovascular responses to increased placental impedance. Pulse wave velocity increased with gestation and was significantly less in the growth-restricted cohort. CONCLUSIONS: Growth restriction is associated with abnormal ventriculovascular physiology that represents a successful adaptive response to raised placental impedance and reduction in wall stress as evidenced by the lower fetal pulse wave velocity in growth-restricted fetuses. However, whilst fetal adaptive mechanisms may aid survival they may result in cerebral and vascular abnormalities that prejudice later cardiovascular health. PMID- 11489227 TI - Transvaginal sonography combined with saline contrast sonohysterography in evaluating the uterine cavity in premenopausal patients with abnormal uterine bleeding. AB - OBJECTIVES: To evaluate whether saline contrast sonohysterography (SCSH) adds additional information to that obtained by transvaginal sonography (TVS) for predicting endometrial abnormality in premenopausal patients with abnormal uterine bleeding. PATIENTS AND METHODS: This was a two-center prospective study at a university clinic and a central hospital in Denmark. The uterine cavity was evaluated with TVS and SCSH in 470 premenopausal patients with abnormal uterine bleeding. One hundred and eighty-nine of the patients had operative hysteroscopy or hysterectomy within 4 months which provided a detailed description of the uterine cavity and was used as the true value for exclusion of polyps and submucous myomas. RESULTS: Based on normal endometrial morphology alone, the results for detection of an abnormal uterine cavity were as follows: sensitivities of TVS 0.92, SCSH 0.99; specificities of TVS 0.62, SCSH 0.72; positive predictive values of TVS 0.80, SCSH 0.85; negative predictive values of TVS 0.82, SCSH 0.98. Transvaginal sonography combined with SCSH was superior to TVS for detection of intracavitary abnormalities (McNemar test, P = 0.008). The post-test probability of there being an abnormal cavity after normal findings on TVS alone was 0.18 (0.10-0.32) and after TVS and SCSH it was 0.02 (0.01-0.11). When normal endometrial morphology was combined with an endometrial thickness of < 12 mm for evaluation of all abnormalities including hyperplasia, the diagnostic potential of TVS or SCSH was almost unchanged except for specificities, which were markedly lower (TVS 0.54; SCSH 0.57). In all the patients referred, TVS had a negative predictive value of 0.94 for identification of polyps and myomas when findings at subsequent SCSH were accepted as the true value. Transvaginal sonography reduced the pretest probability of polyps or submucous myomas from 0.35 to a post-test probability of 0.06, but missed 21% of the polyps. CONCLUSIONS: Sonohysterography was a sensitive tool and was superior to TVS used alone for evaluation of the uterine cavity in patients who underwent operative surgery for abnormal uterine bleeding. All abnormalities except one were found at SCSH, while TVS alone missed polyps and had almost one in four equivocal findings. The use of TVS, without saline contrast, left one in five of the polyps undiagnosed in referred patients with abnormal bleeding. PMID- 11489228 TI - Fetal cardiac effects of doxorubicin therapy for carcinoma of the breast during pregnancy: case report and review of the literature. AB - Cardiotoxicity is a recognized complication of anthracycline drugs given as part of chemotherapy; however, the pre- and postnatal cardiac effects of in utero exposure are not well documented. In this report we present a case of gestational breast cancer with initiation of four cycles of doxorubicin/cyclophosphamide chemotherapy after modified radical mastectomy and axilla dissection during the early second trimester. Serial echocardiographic measurements of the ventricular shortening fraction and biometry of the ventricular cavities were performed. Allowing for the individual variability of these values in the fetus no myocardial dysfunction was observed. The literature was reviewed in an attempt to delineate the possible role of prenatal echocardiography in the diagnosis of doxorubicin-induced cardiotoxicity in the fetus. PMID- 11489229 TI - Sonographic prenatal diagnosis of intracranial fetus in fetu. AB - A case of sonographic prenatal diagnosis of a complex intracranial mass, with features of a fetus in fetu at 17 weeks' gestation, is reported. This diagnosis is reserved for a highly organized tumor containing a vertebral column and recognizable fetal parts and should be differentiated from a teratoma. PMID- 11489231 TI - Fetus in fetu: a rare cause of a neonatal mass. AB - Fetus in fetu is an unusual digression from the normal twinning process which often presents as a mass in neonates. Approximately 80 reports have been documented in the literature to date. The increasing use of routine obstetric ultrasound has identified more cases during the antenatal period. This allows more time for both the neonatology team and the parents to develop a coordinated treatment plan for the anticipated child. We report a case of a fetus in fetu which was first seen as a complex cystic mass during antenatal ultrasound at 21 weeks and was confirmed by subsequent imaging and surgical treatment shortly after birth. PMID- 11489230 TI - Ultrasound prenatal diagnosis of fetus in fetu. AB - A case of fetus in fetu was diagnosed prenatally using ultrasound. The differential diagnosis between a fetus in fetu and a highly differentiated teratoma is discussed. The importance of prenatal diagnosis of fetus in fetu and the effect on subsequent management are described. PMID- 11489232 TI - Prenatal detection of Fraser syndrome without cryptophthalmos: case report and review of the literature. AB - Fraser syndrome (cryptophthalmos-syndactyly syndrome) is an autosomal recessive multiple malformation syndrome whose major manifestations are cryptophthalmos, syndactyly, laryngeal atresia and urogenital defects. Enlarged hyperechogenic lungs contrasted by oligohydramnios, non-visualization of the kidneys and microphthalmia were sonographic markers leading to the prenatal detection of this rare autosomal recessive disorder in earlier reports. We report a case of Fraser syndrome diagnosed at 16 weeks' gestational age in a woman whose previous pregnancy was terminated because of multiple fetal malformations. Abnormal sonographic findings included bilateral agenesis of the kidneys, dilated trachea and main bronchi (suggestive of high airway obstruction), hyperechogenic lungs, syndactyly of the fingers, hepatomegaly, oligohydramnios and hydrops placentae. Face and cerebral structures appeared normal. These findings together with those of the previously affected child led to the diagnosis of Fraser syndrome. The parents elected to terminate the pregnancy. Autopsy results were confirmatory. In conclusion, prenatal diagnosis of Fraser syndrome is possible in the hands of an expert, but due to the great variety of possible malformations the diagnosis will remain doubtful in most cases in which no previous child is affected. PMID- 11489233 TI - Three-dimensional sonographic aspects in the antenatal diagnosis of achondroplasia. PMID- 11489234 TI - Re: Trisomy 21: 91% detection rate using second-trimester ultrasound markers. PMID- 11489235 TI - Thanatophoric dysplasia. PMID- 11489236 TI - Is it good clinical judgment or selection bias? PMID- 11489237 TI - Assessing the quality of clinical trials: can it be done? PMID- 11489238 TI - Acknowledgment of uncertainty: a fundamental means to ensure scientific and ethical validity in clinical research. AB - Recognition of the importance of uncertainty in the design of randomized, controlled trials (RCT) has reached the status of a principle. The "uncertainty principle," or less ambiguously, equipoise, holds that a patient should be enrolled in an RCT only if there is substantial uncertainty about which of the trial treatments would benefit the patient most. In fact, the "uncertainty principle" addresses the most important issue of a clinical trial--the choice of an adequate comparative control. Studies in which intervention and control group are believed to be non-equivalent violate the uncertainty principle. Therefore, one would expect that both editors and authors would be particularly careful to include a statement concerning prior beliefs of the investigator(s) about the uncertainty of the treatments that are reported. However, we found no evidence of such a policy in the randomized, controlled trials we examined. We also show that there is a predictable relationship between the uncertainty principle, that is, the moral principle upon which trials are based, and the ultimate outcomes of clinical trials. We postulate that about 50% of innovations are successful, leading to the conclusion that preserving the ethics of clinical research may be the best investment strategy available. PMID- 11489240 TI - Is cancer care best at high-volume providers? AB - For a variety of medical conditions and procedures, a higher volume-better outcome relationship has been hypothesized for over 25 years. An extensive, consistent body of literature supports a relationship between hospital volume and short-term outcomes for cancers treated with technologically complex surgical procedures. For cancer primarily treated by low-risk surgery, there are few studies. Recent studies found a modest (about 2%) difference in survival benefit between high-volume and low-volume providers associated with colon cancer surgery. Few evaluations in the last 15 years have addressed nonsurgical cancers, eg, lymphomas and testicular cancer. No reports have addressed recurrent or metastatic cancer. Care is better at high-volume providers for a select minority of cancers. Whether provider volume matters in the majority of cancers at the time of presentation has not been evaluated. PMID- 11489239 TI - Economic analysis of randomized, controlled trials. AB - To facilitate the comparison of different treatment strategies, measures have been developed that bring together clinical, quality-of-life, and economic outcomes into summary measures such as the quality-adjusted life year, cost effectiveness, and cost-utility ratios. A number of different types of economic evaluations have been developed, including cost-minimization, cost-effectiveness, and cost-utility analyses. Performance of economic analyses in association with randomized, controlled trials (RCT) has gained increasing enthusiasm in recent years. However, economic measures in RCTs are often outcomes of secondary interest and associated with frequent missing data and inadequate sample size. Variability in the cost measures used and the lack of agreement on clinically meaningful cost differences further limit the conclusions derived from such studies. Economic analyses should be limited to large trials with important trade offs between efficacy and cost. The strengths and limitations of such analyses are discussed, and guidelines are offered for proper economic analyses in randomized, controlled trials. PMID- 11489241 TI - Conflict of interest in industry-sponsored economic evaluations: real or imagined? AB - As industry sponsorship of pharmacoeconomic studies has increased, concerns have been raised about potential biases, owing to the conflicts of interest that this sponsorship creates. A review of the literature indicates that there are some causes for concern, given the fact that most pharmacoeconomic studies report positive findings for the sponsor's drug. However, a more detailed analysis suggests that, although the methodologic quality of some published studies may be poor, the main reason for positive results is that companies only sponsor economic studies where a positive outcome is likely. Therefore, it is concluded that the best way of dealing with perceptions of sponsorship bias is not increased rhetoric, but rather increased public funding for economic evaluation of medicines, thereby creating a true mixed economy for research funding in this field. PMID- 11489242 TI - Combination therapy in hormone-refractory prostate cancer. PMID- 11489243 TI - Immunotherapy for prostate cancer. AB - The components of an effective immune response have been elucidated in recent years. An understanding of the dysfunction of the immune response in cancer in one or more of these components has led to a variety of immunotherapeutic approaches. These therapeutic strategies are designed to stimulate dendritic cell proliferation, promote antigen uptake and processing, stimulate an effector cell response via direct antigen presentation, or target tumor cells via antibody therapy. Many approaches in prostate cancer have demonstrated successful induction of the desired immune response. Limited clinical success has also been seen. PMID- 11489244 TI - The role of resection for patients with renal carcinoma. AB - Metastatic renal cancer is responsive in some cases to immunotherapeutic agents. Indications for nephrectomy in the face of metastatic disease have traditionally included palliation of symptoms caused by the primary tumor, and nephrectomy combined with metastatectomy in patients with resectable metastases. Recent findings from a Southwest Oncology Group trial strongly suggest that cytoreductive nephrectomy, combined with immunotherapy, may also result in improved survival in patients with unresectable metastases. PMID- 11489245 TI - Allogeneic stem-cell transplantation in renal-cell carcinoma. AB - Metastatic renal-cell carcinoma (RCC) remains resistant to nearly all standard cytotoxic therapies, but immune-based cytokine therapies benefit a small minority of patients with advanced RCC. Nonmyeloablative allogeneic stem-cell transplantation is a novel approach to harnessing the immune system to combat this cancer. The strategy relies on a T-cell graft-versus-malignancy effect mediated by donor T cells. Preliminary work in using nonmyeloablative allogeneic stem-cell transplant in RCC has identified a graft-versus-RCC effect and yielded encouraging clinical responses. PMID- 11489246 TI - Gene therapy for prostate cancer. AB - Basic research continues to unravel the molecular complexity of normal and abnormal biologic processes. The development of means to affect the expression level of genes that promote or contribute to cellular transformation, invasion, and metastasis has spawned the concept of gene therapy. This relatively new field seeks to reverse or suspend the pathologic progression of a variety of diseases including the malignant transformation of prostatic epithelial cells. Initial clinical trials for prostate cancer have thus far shown gene therapy to be relatively safe, although definitive evidence of durable therapeutic efficacy remains to be demonstrated. In this article, recent preclinical research, current therapeutic strategies, and recent results of gene therapy clinical trials for the treatment of prostate cancer are reviewed. PMID- 11489247 TI - Complementary and alternative medicine in prostate cancer. AB - Prostate cancer patients, like other cancer patients as well as the general population, are increasingly exploring the use of complementary and alternative medicine (CAM). This paper describes the use of CAM in this patient population and the evidence regarding some CAM treatments in the setting of prostate cancer. Some herbal agents and micronutrients have demonstrated biologic activity that may benefit patients with prostate cancer. The clinical effects of these and others and the potential interactions among CAM treatments and with conventional treatment remain an appropriate target for further investigation. PMID- 11489248 TI - Influence of sleep deprivation coupled with administration of melatonin on the ultrastructure of rat pineal gland. AB - The effects of sleep deprivation with or without melatonin treatment on the pineal morphology in rats were studied. Five days after sleep deprivation and using electron microscopy, many of the pinealocytes exhibited structural alterations including dilation of the cisternae of the rough/smooth endoplasmic reticulum, Golgi saccules and mitochondria, and an increase in the numbers of lipid droplets, vacuoles and dense-core vesicles. These features were considered as morphological evidence of increased synthesis or secretion by the pineal gland. In addition, numerous membranous profiles, considered to be degraded cellular organelles, were observed in some pinealocytes and sympathetic nerve terminals. It is suggested that the occurrence of degenerating organelles had resulted from the deleterious effect of sleep deprivation. This may be attributed to an overload of secretory activity of the pineal gland during stress elicited by the long-term sleep deprivation, leading to functional exhaustion and irreversible damage of the oxidation-related organelles. In sleep-deprived rats receiving a single injection of melatonin (10 mg/kg) for 5 consecutive days, the above features indicative of pinealocytic activation were attenuated. In fact, all signs of degeneration of cellular organelles were rarely found. These results suggest that the pineal gland is itself a target for exogenously administered melatonin. Thus, melatonin when administered systemically may be used as a potential neuroprotective drug against neuronal damage induced by sleep deprivation. PMID- 11489249 TI - MK-801 reduces non-noxious stimulus-evoked Fos-like immunoreactivity in the spinal cord of rats with chronic constriction nerve injury. AB - We investigated the role of N-methyl-D-aspartate (NMDA) receptors on non-noxious stimulus-induced pain by examining the effect of MK-801, a non-competitive NMDA receptor antagonist, on Fos-like immunoreactivity (FLI) in the spinal dorsal horn by non-noxious stimulation to rats with chronic constriction injury (CCI) of the sciatic nerve. In CCI rats that did not receive the non-noxious stimulus, FLI was significantly increased in laminae V/VI of the dorsal horn at the 7th and 14th days after surgery relative to sham rats. When CCI rats received non-noxious stimuli, rubbing the plantar of the hind paw, FLI in laminae I/II at the 14th day was significantly increased relative to CCI rats that did not receive the stimulation. In sham rats, the same stimulus significantly decreased FLI in laminae III/IV and V/VI at the 7th and 14th day. When MK-801 was administered intraperitoneally prior to non-noxious stimulation in CCI rats at the 14th day after surgery, the stimulus-induced FLI in laminae I/II in CCI rats was significantly reduced. This study indicates that NMDA receptor is involved in upregulating FLI in response to non-noxious stimulation of CCI rats. PMID- 11489250 TI - Vasopressin induced cyclooxygenase dependent superoxide generation contributes to K(+) channel function impairment after brain injury. AB - This study determined if vasopressin generates superoxide anion (O2(-)) in a cyclooxygenase dependent manner and if such production contributes to impairment of dilation to activators of ATP sensitive K(+) (K(ATP)) and calcium sensitive K(+) (K(ca)) channels following fluid percussion brain injury (FPI) in newborn pigs equipped with closed cranial windows. Superoxide dismutase (SOD) inhibitable nitroblue tetrazolium (NBT) reduction was determined as an index of O2(-) generation. Under non-brain injury conditions, topical vasopressin (40 pg/ml, the concentration present in CSF following FPI) increased SOD inhibitable NBT reduction from 1+/-1 to 25+/-4 pmol/mm(2). Indomethacin, a cyclooxygenase inhibitor, blunted such NBT reduction (1+/-1 to 5+/-1 pmol/mm(2)), while the vasopressin antagonist, l-(beta-mercapto-beta beta-cyclopentamethylene propionic acid) 2-(o-methyl)-Tyr-AVP (MEAVP) blocked NBT reduction. MEAVP and indomethacin also blunted the NBT reduction observed after FPI. Under non-brain injury conditions, vasopressin (40 pg/ml) coadministered with the K(ATP) and K(ca) channel agonists, cromakalim and NS1619 (10(-8), 10(-6) M) diminished dilation to these K(+) channel agonists while indomethacin partially prevented such impairment (13+/-1 and 23+/-1 vs. 4+/-1 and 10+/-2 vs. 8+/-1 and 19+/-1% for cromakalim in untreated, vasopressin, and vasopressin plus indomethacin treated piglets, respectively). Cromakalim and NS1619 induced pial artery dilation was attenuated following FPI, while indomethacin or MEAVP preadministration partially prevented such impairment (13+/-1 and 23+/-1, sham control; 1+/-1 and 4+/-1, FPI; 8+/-1 and 16+/-3%, FPI-indomethacin pretreated for responses to cromakalim 10( 8), 10(-6) M, respectively). These data show that vasopressin increased O2(-) production in a cyclooxygenase dependent manner and contributed to this production after FPI. These data also show that vasopressin blunted K(ATP) and K(ca) channel mediated cerebrovasodilation in a cyclooxygenase dependent manner. These data suggest that vasopressin induced cyclooxygenase dependent O2(-) generation contributes to K(ATP) and K(ca) channel function impairment after FPI. PMID- 11489251 TI - Identification of dopamine responsive mRNAs in glial cells by suppression subtractive hybridization. AB - Recent studies have established that glial cells are important targets of the neurotransmitter dopamine (DA), but the regulatory effects of DA on glial cells have not been extensively studied. In the present study, we have investigated the influence of DA on gene transcription in glial cells. Two-directional (forward and backward) suppression subtraction hybridization (SSH) was performed on astrocytes cultured from rat cerebral tissues in standard media or in culture media treated with DA. PCR-select differential screening was used to further verify the differentially expressed cDNA clones, positive clones were sequenced, and the mRNAs were re-examined on Northern blots. Fourteen sequences were identified of which eleven are homologous to known genes, three are homologous to expressed sequence tags (ESTs). Three novel full-length cDNAs were isolated using the EST fragments as probes to screen a cDNA library constructed from human brain. Analysis of these sequences suggested that complex intracellular signaling pathways, involving crosstalk with growth factor pathways, steroid hormone pathways, and an interferon-regulated 2-5 A pathway, are responsive to DA in astrocytes. The responsive proteins downstream from the signaling pathways were found to fall into at least three groups, including a series of metabolic enzymes, stress proteins, transfer proteins, etc. In addition, several of them have established their relationships with specific neurodegenerative diseases, showing that there is overlap in the pathogenic mechanisms of different diseases. Our results have provided a foundation for better understanding of the molecular basis of glial cell functions in dopaminergic transmission and an approach to find possible medication for the related disorders. PMID- 11489252 TI - The distribution and characterization of NADPH-d/NOS-IR neurons in the rat cuneate nucleus. AB - Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry have been used to characterize the nitric oxide (NO)-containing neurons in the rat cuneate nucleus. The present results showed that NADPH-d-positive/NOS-immunoreactive (-IR) neurons were distributed in the entire rostrocaudal extent of the nucleus. In the caudal region (approximately 1-2 mm caudal to the obex), NADPH-d/NOS-IR neurons were aggregated along the dorsal area of the nucleus notably in the lateral aspect. When traced rostrally, labeled neurons were progressively reduced and the cells were randomly distributed. The labeled neurons varied from round, ovoid to spindle-shaped with a mean profile area of about 140.1+/-1.7 microm(2) (n=720). They made up 7-10% of the neuronal population in the cuneate nucleus. By immunoelectron microscopy, the immunoreaction product was deposited throughout the cytoplasm extending from the soma to the proximal and distal dendrites. Results of NADPH-d staining paralleled that of NOS immunohistochemistry. Furthermore, NADPH-d reactivity and NOS-IR were colocalized in the same neurons following double labeling. Using NADPH-d histochemistry along with anti-gamma aminobutyric acid (GABA) and -glycine postembedding immunolabeling for identification of GABA- and glycine-IR neurons, respectively, about 33% of the NADPH-d-positive neurons contained both GABA and glycine, 26% of them contained only glycine, while 41% of them showed neither GABA nor glycine labeling. Cuneothalamic neurons (CTNs) were identified by injecting the retrograde tracer Fluorogold (FG) into the ventrobasal complex of the thalamus. Numerous FG-labeled neurons were present in the contralateral cuneate nucleus, but none were reactive for NADPH-d. The present results suggest that approximately 60% of the NADPH d/NOS-IR neurons in the cuneate nucleus are interneurons containing GABA and/or glycine. PMID- 11489253 TI - Extracellular serotonin variations during vigilance states in the preoptic area of rats: a microdialysis study. AB - Numerous studies have shown that serotonergic transmission decreases from waking (W) to slow wave sleep (SWS) to paradoxical sleep (PS), suggesting an active role of serotonin (5-HT) in W but not in sleep. Conversely, the inhibition of 5-HT activity produces insomnia. This insomnia can be reversed by injections of 5 hydroxytryptophan in the preoptic area (POA), suggesting that 5-HT is necessary in this cerebral structure for sleep. Using microdialysis, we studied, 5-HT variations in the POA of rats in relation to vigilance states. 5-HT levels were higher during W than during during SWS and PS. 5-HT increased just before the rats fell asleep and then decreased during sleep. A decreased 5-HT transmission was also observed from SWS to PS. These data document a positive correlation between 5-HT levels in POA and wakefulness. Moreover, these observations are in favour of a permissive role of 5-HT in the POA during PS. A comparison between the POA and the prefrontal cortex in the sleep-wake cycle is discussed. PMID- 11489254 TI - Short-term exposure to a neuroactive steroid increases alpha4 GABA(A) receptor subunit levels in association with increased anxiety in the female rat. AB - Previous work from this laboratory has demonstrated that withdrawal from the neuroactive steroid 3alpha,5alpha-THP (3alpha-hydroxy-5alpha-pregnan-20-one) after 3-week exposure to its parent compound, progesterone (P), increases anxiety and produces benzodiazepine (BDZ) insensitivity in female rats. These events were linked to upregulation of the alpha4 subunit of the GABA(A) receptor (GABAR) in the hippocampus [Brain Res. 507 (1998) 91; Nature 392 (1998) 926; J. Neurosci. 18 (1998) 5275]. The present study investigates the role of shorter term hormone treatment on alpha4 subunit levels as well as relevant behavioral and pharmacological end-points related to GABAR function. After 2-3 days of P exposure, two- to threefold increases in alpha4 protein levels were observed, which declined to control values after 5-6 days of hormone exposure. This effect was due to the GABA-modulatory metabolite of P, 3alpha,5alpha-THP. alpha4 upregulation was inversely correlated with BDZ potentiation of GABA-gated current, assessed using whole cell patch clamp techniques on acutely isolated hippocampal pyramidal cells. A near total BDZ insensitivity was observed by 2-3 days of hormone exposure in association with the maximal increase in alpha4 levels. Up-regulation of the alpha4 GABAR subunit was also reflected by an increase in anxiety in the elevated plus maze. A significant decrease in open arm entries was observed after 72-h exposure to P, an effect which recovered by 6 days of P treatment. As demonstrated in vitro, alpha4 upregulation also resulted in a relative insensitivity to the anxiolytic actions of BDZ. These results suggest that short-term exposure to 3alpha,5alpha-THP produces changes in GABAR subunit composition similar to those that occur after chronic exposure and withdrawal from the steroid. PMID- 11489255 TI - Atropine-sensitive and -insensitive components of the somatosensory evoked potential. AB - The evoked potential in primary somatosensory cortex changes with time. Short puffs of air administered to the nose of awake, quietly resting adult rats elicited potentials that could be altered by one of several treatments (saline, atropine methyl nitrate or atropine sulfate). The change produced by blocking muscarinic receptors in the central nervous system with atropine sulfate (100 mg/kg) was the largest, but control substances also altered the potential, suggesting that the gradual changes observed in the evoked potential 30 min after intraperitoneal injection may also be affected by factors such as the stress associated with injection itself and the blockade of peripheral muscarinic receptors. The changes observed in the evoked potential when central cholinergic receptors are blocked include a large shift towards positivity in the early components (between 18 and 64 ms with maxima at 20 and 47 ms) and a similarly significant shift towards negativity in the later components (between 90 and 208 ms with maxima at 115 and 157 ms). The actual changes observed during inactivation of central muscarinic receptors suggest that the role of acetylcholine during arousal is more than to simply bias the cortex towards greater excitability. Rather, the muscarinic receptors on inhibitory interneurons or on the dendritic terminals of pyramidal cells in superficial layers of cortex enhance the first intracortical synaptic events but reduce the population response at later times during the first 250 ms following a tactile stimulus. PMID- 11489256 TI - Rapid differential conditioning of the somatosensory evoked potential by changed patterns of brief innocuous tactile stimuli in waking rats is altered by atropine sulfate. AB - Air puffs delivered to the nose of an awake, lightly restrained rat every 15 s produced evoked potentials that changed gradually over time so that the averaged response to the last 40 stimuli was measurably different from the first 40. This habituation-like paradigm increased the size of an early component of the potential in several places. When measured with respect to the time of stimulus onset (there was a 21.6 ms delay in the time of arrival of the stimulus maximum at the nose), one of the largest increases occurred 46 ms later (39 ms latency to onset, and 55 ms latency to offset). As well, a late component of the waveform became more positive, showing a maximum between 156 and 185 ms (133 ms latency to onset, and more than 250 ms latency to offset). Changing the pattern but not the number of stimuli accelerated the rate of this positive shift with a maximum at 37 ms (21 ms latency to onset, and 42 ms latency to offset), but did not affect the rate of change in the late component. This effect of altering the temporal pattern of the stimuli was blocked by systemic injections of atropine sulfate, a blocker of central muscarinic receptors, whereas, neither saline injections nor atropine methyl nitrate injections (an atropine analog that does not cross the blood-brain barrier) could produce these changes. These observations suggest that the adaptive changes of the somatosensory evoked potential induced by novel patterns intercalated in otherwise monotonous repetitive somatic stimuli depend upon central muscarinic mechanisms. PMID- 11489257 TI - Regional variation in brain capillary density and vascular response to ischemia. AB - Differences in brain neuroarchitecture have been extensively studied and recent results demonstrated that regional differences in the physiological properties of glial cells are equally common. Relatively little is known on the topographic differences in vascular supply, distribution and density of brain capillaries in different CNS regions. We developed a simple method consisting of intravascular injection of fluorescent dyes coupled to immunocytochemical techniques that allows for simultaneous observation of glia-neuronal-vascular interactions in immersion-fixed brain specimens from small rodents. This technique permits quantitative evaluation of regional differences in glial/neuronal distribution and the study of their relationship to vascular densities. Variations of this technique also allow the detection of abnormal microvasculature (i.e. 'leaky' vessels), a useful feature for studies of blood-brain barrier function in health and disease. By use of quantitative confocal microscopy, the three-dimensional geometry of cortical and hippocampal structures revealed remarkable differences in vascularization between cortical gray/white matter junction, and hippocampal formation (CA1 and CA3 regions). Significant differences were also observed within the same investigative region: CA1 was characterized by low capillary density compared to neighboring CA3. Following an ischemic insult, CA1 vessels had more extensive blood-brain barrier leakage than CA3 vessels. We conclude that in addition to neuronal and glial heterogeneity, cortical structures are also endowed with region-specific vascular patterns characterized by distinct pathophysiological responses. PMID- 11489258 TI - Adenosine receptor antagonists cancelled the ischemic tolerance phenomenon in gerbil. AB - Pretreatment of the brain with sublethal ischemia has been reported to induce neuronal resistance to otherwise lethal ischemia, a phenomenon designated as ischemic tolerance. The protective mechanisms of the phenomenon are not known yet, however, recent experimental data suggest the involvement of adenosine receptor activation in the acquisition of tolerance. In this study, the effect of theophylline, a non-selective adenosine receptor antagonist, and 8-cyclopentyl 1,3-dipropylxanthine (DPCPX), an adenosine A1 receptor antagonist, were investigated to ascertain if these drugs could cancel the protective effect of ischemic tolerance in the gerbil. DPCPX or theophylline was administered at 3 h after a short preconditioning ischemia, and 21 h later animals were subjected to lethal ischemia of 5 min duration. DPCPX at a dose of 1.0 mg/kg (i.p) and theophylline at a dose of 20 mg/kg (i.p) significantly reduced the protective effect of preconditioning in the CA1 hippocampal neurons. These findings suggest the involvement of adenosine receptor activation for the development of ischemic tolerance phenomenon. PMID- 11489259 TI - Longitudinal analysis of motor activity and coordination, anxiety, and spatial learning in mice with altered blood pressure. AB - Mice with either high or low blood pressure (BP) were compared to normotensive controls at 2 and 12 months of age for motor activity, equilibrium, anxiety, and spatial learning. Irrespective of age, high BP mice were more active in an open field than normotensive controls, whereas low BP mice were hypoactive at 2 months of age. High BP mice had a higher number of entries and a longer duration of visits in the open arms, a higher open arm/total arm ratio, a longer duration for the first visit into an open arm, and lower latencies before entering the first open arm than controls in the elevated +-maze, indicative of reduced anxiety. Reduced levels of anxiety were also displayed by low BP mice for the duration of the first open arm visit (both age groups) and for the time spent in the open arms (older group). In the motor coordination test (coat-hanger), high BP mice had higher two-paw movement time and reached the top of the apparatus on fewer occasions than controls. Both groups with abnormal BP values were deficient during visuomotor guidance in the water maze. These results indicate strain-, age , and test-specific abnormalities in mice with uncontrolled hypertension or hypotension. PMID- 11489260 TI - Altered gene expression in cerebral capillaries of stroke-prone spontaneously hypertensive rats. AB - Stroke-prone spontaneously hypertensive rats (SHRSP) are a well-characterized, genetic model for stroke. We showed earlier that the structure and function of the tight junctions in SHRSP blood-brain barrier endothelial cells is disturbed prior to stroke. To investigate the molecular events leading to endothelial dysfunction in SHRSP cerebral capillaries, we carried out suppression subtractive hybridization (SSH) in combination with a cDNA filter screening step. We identified two cDNA fragments that were upregulated in SHRSP, compared to stroke resistant spontaneously hypertensive rats (SHR), and found open reading frames of 133 and 138 amino acids, respectively. These peptides did not match any known proteins in public databases. A third upregulated SHRSP cDNA fragment was identified as the rat sulfonylurea receptor 2B (SUR2B). We also isolated and cloned the cDNA of the rat homologue for the mouse G-protein signaling 5 (RGS5) regulator. This regulator was downregulated in SHRSP. We used in situ hybridization to show that rat RGS5 is expressed in the brain capillary endothelium and in the choroid plexus. Our findings may lead to the identification of new stroke-related genes. PMID- 11489261 TI - Interactions of 1-methyl-4-phenylpyridinium and other compounds with P glycoprotein: relevance to toxicity of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. AB - The vesicular monoamine transporter 2 (VMAT2) has sequence homology with bacterial multidrug transporters which in turn share homology with mammalian P glycoprotein (P-GP). Both VMAT2 and P-GP can detoxify cells. 1-Methyl-4 phenylpyridinium (MPP(+)), the toxic metabolite of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), is a substrate for VMAT2 that has several structural features in common with P-GP substrates and inhibitors. The present studies investigated whether P-GP is responsible for the elimination of MPP(+) from the brain. Additionally, VMAT2 and P-GP are inhibited by many of the same compounds. Thus we also investigated whether VMAT2 inhibitors could block P-GP in vitro and vice versa whether P-GP inhibitors could block VMAT2 mediated transport of [3H] DA into synaptic vesicles. In mice treated with MPTP and a P-GP inhibitor (quinidine, trans-flupentixol or cyclosporine A), the elimination of MPP(+) from the striatum was significantly delayed. However, in experiments using various cell lines expressing either mouse or human P-GP, MPP(+) did not reverse the P-GP mediated resistance to vincristine, suggesting that MPP(+) is a poor substrate for P-GP. Additional experiments were performed using mdr1a/b double knockout mice which lack functional P-GP encoded by these two genes. Data from mdr1a/b knockout mice treated with MPTP also suggest that MPP(+) is not extruded from the brain by P-GP. In other studies, we demonstrated that the VMAT2 inhibitors tetrabenazine and Ro 4-1284 inhibit P-GP and that the P-GP inhibitors trans flupentixol and quinidine inhibit VMAT2. Thus, several new drugs can be added to the list of compounds that are able to inhibit both VMAT2 and P-GP, providing further evidence of the similarity between these two transporters. PMID- 11489262 TI - Cytochrome oxidase activity in the olfactory system of staggerer mutant mice. AB - The staggerer mutation is characterized by the disruption of a single recessive gene encoding for the nuclear hormone receptor RORalpha. In addition to the well studied gene expression in the cerebellum causing massive primary Purkinje cell loss, the RORalpha gene is also expressed in the thalamus and the olfactory bulb. A quantitative histochemical study of cytochrome oxidase activity was performed in staggerer mutants and their respective controls in order to determine whether olfactory bulb neuropathology leads to neuronal metabolic alterations in olfactory and related limbic regions. In the staggerer olfactory bulb, the core and the shell of the glomeruli had lower levels of cytochrome activity, whereas higher levels were found in the external plexiform and granular layers. Other olfactory and limbic regions were unchanged, except for a higher level in the accessory olfactory bulb and a lower level in the most ventral part of the medial orbital cortex. These results are discussed with regard to the olfactory deficits and changes in social interactions previously observed in this mutant. PMID- 11489263 TI - Differentiation of NG108-15 cells induced by the combined presence of dbcAMP and dexamethasone brings about the expression of N and P/Q types of calcium channels and the inhibitory influence of muscarinic receptors on calcium influx. AB - Differentiation of cholinergic cell line NG108-15 induced by a combination of dibutyryl cyclic AMP (dbcAMP) and dexamethasone enhances the cholinergic phenotype of the cells more than that induced by either agent alone. We investigated the effect of treatment with dbcAMP and dexamethasone on potassium depolarization-evoked influx of calcium and its regulation by the muscarinic agonist carbachol. Depolarization of control cells and of cells differentiated in the presence of dbcAMP or dexamethasone alone, or in the combined presence of dbcAMP and dexamethasone induced, respectively, 2.2-, 4.3-, 2.7- and 10.7-fold increases of the resting [Ca(2+)](i). Dexamethasone alone and the combination of dbcAMP and dexamethasone augmented the number of muscarinic receptors by 25 and 40%, respectively. Inhibitors of N (omega-conotoxin GVIA) or P/Q (omega-agatoxin TK) calcium channels had no effect on Ca(2+) influx in control cells, whereas in cells differentiated in the combined presence of dbcAMP and dexamethasone they significantly diminished the influx of Ca(2+) by 20 and 5%, respectively. Carbachol attenuated calcium influx in differentiated cells in an atropine insensitive manner if it was present during stimulation. This effect of carbachol was probably due to an open-channel block of L type channels. In the presence of nifedipine, carbachol attenuated the influx of Ca(2+) into cells differentiated with dbcAMP and dexamethasone by 20% in an atropine-sensitive way. Data show that differentiation of NG108-15 cells by dbcAMP and dexamethasone promotes the expression of functional nifedipine-insensitive N and P/Q types of Ca(2+) channels and that the nifedipine-insensitive calcium influx becomes subject to inhibitory regulation by muscarinic receptors. PMID- 11489264 TI - Somatotopic reorganization in the brainstem and thalamus following peripheral nerve injury in adult primates. AB - Injury-induced reorganization of central somatotopic maps is a phenomenon that has proven to be useful for elucidating the mechanisms and time course of neural plasticity. To date, the overwhelming majority of this line of research has focused on such plastic events in cortical areas, at the expense of subcortical structures. In this study, we used multi-unit electrophysiological recording techniques to assess the somatotopic organization of brainstem and thalamic areas following chronic survival from paired median and ulnar nerve section in adult squirrel monkeys. We report that the extent of cutaneously-driven reorganization in both the cuneate nucleus of the brainstem and the ventroposterior lateral nucleus of the thalamus is comparable to that previously documented for area 3b of cortex. These observations are consistent with those previously reported in thalamus, and are unique for brainstem. PMID- 11489265 TI - Dual effects of interleukin-1beta on N-methyl-D-aspartate-induced retinal neuronal death in rat eyes. AB - In this study we determine if interleukin-1beta (IL-1beta) modulates N-methyl-D aspartate (NMDA)-induced retinal damage. Sprague-Dawley rats were anesthetized with inhalation of halothane, after which a single injection of 5 microl of IL 1beta (0.1 to 10 ng/eye) (and/or IL-1 receptor antagonist (IL-1ra)) for experimental eyes was administered. Two days later (or simultaneously), NMDA (20 nmol) was injected into the vitreous space. One week later, each eye was enucleated and transverse sections were subjected to morphometric analysis. Enzyme-linked immunosorbent assay (ELISA) was conducted for the determination of IL-1beta levels in retina. Immunohistochemical and immunoblot studies were also performed. In eyes that received an intravitreal injection of IL-1beta (0.1 to 10 ng/eye), significant thinning of the inner plexiform layer (IPL) was observed (P<0.05). Immunohistochemical and ELISA studies demonstrated upregulated expression of IL-1beta in retinas that had undergone NMDA injection. Treatment with 10 ng of IL-1ra induced a protective effect against NMDA-induced retinal damage. Pretreatment with IL-1beta induced a significant protective effect on NMDA-induced retinal damage. Our studies suggest that IL-1beta induces neuronal cell death directly, as shown by the protective effects of IL-1ra, but has a protective effect on NMDA-induced retinal damage indirectly after an incubation time of at least 2 days. PMID- 11489266 TI - Cyclooxygenase-2 mediates the febrile response of mice to interleukin-1beta. AB - Various lines of evidence have implicated cyclooxygenase (COX)-2 as a modulator of the fever induced by the exogenous pyrogen lipopolysaccharide (LPS). Thus, treatment with specific inhibitors of COX-2 suppresses the febrile response without affecting basal body (core) temperature (T(c)). Furthermore, COX-2 gene ablated mice are unable to develop a febrile response to intraperitoneal (i.p.) LPS, whereas their COX-1-deficient counterparts produce fevers not different from their wild-type (WT) controls. To extend the apparently critical role of COX-2 for LPS-induced fevers to fevers produced by endogenous pyrogens, we studied the thermal responses of COX-1- and COX-2 congenitally deficient mice to i.p. and intracerebroventricular (i.c.v.) injections of recombinant murine (rm) interleukin (IL)-1beta. We also assessed the effects of one selective COX-1 inhibitor, SC-560, and two selective COX-2 inhibitors, nimesulide (NIM) and dimethylfuranone (DFU), on the febrile responses of WT and COX-1(-/-) mice to LPS and rmIL-1beta, i.p. Finally, we verified the integrity of the animals' responses to PGE2, i.c.v. I.p. and i.c.v. rmIL-1beta induced similar fevers in WT and COX-1 knockout mice, but provoked no rise in the T(c)s of COX-2 null mutants. The fever produced in WT mice by i.p. LPS was not affected by SC-560, but it was attenuated and abolished by NIM and DFU, respectively, while that caused by i.p. rmIL-1beta was converted into a T(c) fall by DFU. There were no differences in the responses to i.c.v. PGE2 among the WT and COX knockout mice. These results, therefore, further support the notion that the production of PGE2 in response to pyrogens is critically dependent on COX-2 expression. PMID- 11489267 TI - Evaluation of neurodegeneration in scrapie-infected animals by selective methods that detect cellular degeneration. AB - Scrapie is a fatal neurodegenerative disease of sheep and goats. The precise details of neuronal and neurite degeneration in scrapie-infected animals remain unknown. Using specific silver staining methods, we compared the neurodegeneration caused by treatment of rats with kainic acid (KA) or ibogaine (IBO) to the neuropathology observed in mice infected with the C602 strain of scrapie. As reported previously, KA resulted in extensive silver labeling of neurons, especially in the cortex, putamen and hippocampus. IBO silver labeling was observed only in small clusters of Purkinje neurons in the paravermal region of the cerebellum. However, in scrapie-infected mice, a few silver stained neurons (differing from the dark degenerating neurons observed following neurotoxic exposure) were found in layer II of cortex, cingulate cortex, zona incerta, thalamus and hypothalamus. Some silver grains were observed in glial like cells, especially those in the paraventricular region. Degenerating axons were positive for silver staining and were found in the cortex, cingulate cortex, corpus callosum, habenulae, septum, fornix, thalamus, caudate putamen and a few in fasciculus retroflexus and substantia nigra. Our results suggest that the limbic system is one of the important loci for the neurodegenerative effect of at least some scrapie strains. PMID- 11489268 TI - Activation of spinal serotonin(2A/2C) receptors augments nociceptive responses in the rat. AB - The role of spinal 5-HT(2A/2C) receptors in the regulation of spinal nociceptive transmission was studied. The 5-HT(2A/2C) agonist (+/-)-1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane (DOI) and the antagonist ketanserin tartrate were administered intrathecally immediately before the formalin test. Activation of spinal 5-HT(2A/2C) receptors increased the pain-like behavioural response in both the early and late phases. The findings support the hypothesis that spinal 5 HT(2A/2C) receptors augment the spinal afferent nociceptive impulses induced by peripheral inflammation. PMID- 11489269 TI - Effects of ENA713 and CHF2819, two anti-Alzheimer's disease drugs, on rat amino acid levels. AB - The effects of oral ENA713 and CHF2819 (0.5, 1.5 and 4.5 mg/kg), two novel acetylcholinesterase inhibitors, on extracellular concentrations of amino acids in rat hippocampus, were evaluated using in vivo microdialysis. ENA713, at 4.5 mg/kg, but not CHF2819, significantly decreased glutamate, taurine, arginine and citrulline levels, without affecting aspartate concentrations. These results suggest that the modulation of amino acidergic transmission could represent an additional mechanism of action in Alzheimer's disease for some acetylcholinesterase inhibitors. PMID- 11489270 TI - Post-ischemic hypothermia blocks caspase-3 activation in the newborn rat brain after hypoxia-ischemia. AB - The effects of hypothermia on caspase-3 activation were investigated in the newborn rat brain after hypoxia-ischemia (HI). Intense caspase-3 activation was observed in the control brains after HI, but this activation was significantly reduced by postischemic hypothermia. These findings suggest that the inhibition of caspase-3 activation may be an interventional point underlying the neuroprotective effect of hypothermia in neonates. PMID- 11489271 TI - Determination of the extracellular concentration of glycine in the rat spinal cord dorsal horn by quantitative microdialysis. AB - Two quantitative microdialysis methods were used to determine the extracellular concentration of glycine in the dorsal spinal cord of halothane-anaesthetised rats. Extracellular glycine determined by zero net flux was 2.6+/-0.3 microM and by the zero flow method was 3.3+/-0.3 microM. For comparison the glycine content of cerebrospinal fluid was determined to be 6.4+/-1.1 microM. There was no correlation between the extracellular and the cerebrospinal fluid concentrations. PMID- 11489272 TI - Inhibitory effects of tandospirone, a 5-HT1A agonist, on medial vestibular nucleus neurons responding to lateral roll tilt stimulation in rats. AB - An electrophysiological study was performed using chloral hydrate-anesthetized rats to determine whether tandospirone, a 5-HT1A agonist, affects neuronal activities of the medial vestibular nucleus (MVN), since serotonergic innervation and 5-HT1A receptors are present in this nucleus. Tandospirone applied microiontophoretically at a current of 20-60 nA caused an inhibition of tilt induced firing of alpha-type neurons, which showed increased and decreased firing with lateral tilt ipsilateral and contralateral to the recording site, respectively, along with that of beta-type neurons which exhibited the reverse responses to ipsilateral and contralateral tilt stimulation. The inhibition was antagonized during simultaneous, iontophoretic application of WAY-100635 (20-60 nA), a 5-HT1A receptor antagonist, although WAY-100635 alone rarely affected spontaneous or tilt-induced firing in either type of neurons. These results suggest that tandospirone acts on a 5-HT1A receptor to inhibit transmission of otolith information to alpha- and beta-type MVN neurons. PMID- 11489273 TI - Enhanced Ca(2+) influx with mossy fiber stimulation in hippocampal CA3 neurons of spontaneously epileptic rats. AB - This study was performed to determine whether the intracellular Ca(2+) concentration ([Ca(2+)](i)) is increased in hippocampal CA3 neurons of spontaneously epileptic rats (SER) which show both absence-like and convulsive seizures using hippocampal slices loaded with Calcium Green-1 when a weak single stimulation is given to the mossy fiber. [Ca(2+)](i) in the CA3 area was significantly increased after a single stimulus to mossy fibers in SER, while no changes were detected in normal Wistar rats. These findings suggest the existence of an abnormality in the Ca(2+) channel in the SER CA3 region and that this is probably responsible for epileptic seizures. PMID- 11489274 TI - Synaptically-released zinc inhibits N-methyl-D-aspartate receptor activation at recurrent mossy fiber synapses. AB - Hippocampal slices from pilocarpine-treated rats were used to explore the effect of zinc released at mossy fiber synapses on dentate granule cells. Chelation of zinc enhanced the N-methyl-D-aspartate (NMDA) receptor-mediated component of the excitatory postsynaptic current (EPSC), but did not affect the AMPA/kainate receptor-mediated component. Its effect was detectable only at negative membrane potentials and was pathway specific. Thus corelease of zinc reduces the ability of glutamate to activate postsynaptic NMDA receptors. Through this action, zinc would be expected to attenuate granule cell epileptiform activity supported by the recurrent mossy fiber pathway. PMID- 11489277 TI - Pathological study of the diffuse myelin pallor in the anterolateral columns of the spinal cord in amyotrophic lateral sclerosis. AB - An immunohistochemical study using a monoclonal antibody against macrophage (Ki M1p) was performed to examine which fiber tracts are affected in the spinal cords and brainstems of ALS patients. In 21 out of 30 ALS patients, various degrees of macrophage infiltration were observed diffusely in the anterolateral columns beyond the corticospinal tracts. On the other hand, a few macrophages were scattered in 20 non-ALS patients in the anterolateral columns outside the corticospinal tracts. In ALS brainstems, the macrophages were mainly localized in the corticospinal tracts. The result suggests that the diffuse myelin pallor in the anterolateral columns beyond the corticospinal tracts may be derived from intrinsic spinal cord lesions. Quantitative investigation using a monoclonal antibody against phosphorylated neurofilaments (SMI-31) revealed that the decrease in the numbers of small fibers would induce the diffuse myelin pallor in anterolateral columns of ALS patients. From these findings, we propose that the propriospinal bundles are candidates for the degenerating fibers in the anterolateral columns of ALS. PMID- 11489278 TI - Sensorimotor demyelinating neuropathy with IgM antibody against gangliosides GD1a, GT1b and GM3. AB - We report a patient with sensorimotor demyelinating neuropathy with high-titer IgM antibody against gangliosides GD1a, GT1b and GM3. The patient was a 65-year old male who was hospitalized with chief complaints of muscular weakness of all limbs and numbness of the hands and feet. Nerve-conduction studies revealed reduced conduction velocities of the motor nerves with increased temporal dispersion and loss of sensory nerve action potentials. Treatment with steroids was ineffective. IgM antibody against GD1a, GT1b and GM3, which are known to be the ligands for myelin-associated glycoprotein (MAG), might have played a role in the demyelination in this patient by inhibiting adhesion between myelin and axonal membrane. PMID- 11489279 TI - Congenital glioblastoma multiforme: a report of three cases and a review of the literature. AB - The clinical histories, histologic features, and immunohistochemical staining patterns for glial fibrillary acidic protein, vimentin, p53, and epidermal growth factor receptor (EGFR) of three cases of congenital glioblastoma multiforme are given and previous case reports are reviewed. Of the three cases reported in this series, two have had long-term survivals of greater than 2 1/2 and 5 1/2 years after surgery and surgery followed by chemotherapy, respectively. Both of these cases also demonstrated p53 protein accumulation, a finding in pediatric glioblastoma multiforme associated with poor prognosis. The third case occurred in an infant who died at birth and demonstrated a well-circumscribed tumor that did not invade adjacent brain parenchyma. Considering these three cases, the biological behavior in congenital glioblastoma multiforme may not be unfavorable as portrayed in the literature or as seen in its adult counterpart. PMID- 11489280 TI - Molecular genetic analysis of Charcot-Marie-Tooth 1A duplication in Norwegian patients by quantitative photostimulated luminescence imaging. AB - Around 70% of Charcot-Marie-Tooth 1 (CMT1) cases are caused by a dominantly inherited 1.5-Mb duplication at 17p11.2-12 (CMT1A). Using photostimulated luminescence (PSL) imaging of MspI Southern blots, hybridization signals of the probe pVAW409R3a in relation to cohybridized probe SF85a, were densitometrically quantified and an RFLP allele-band ratio determined. A total of 55 Norwegian CMT patients and 16 asymptomatic family members from 26 separate families, clinically and neurophysiologically classified as CMT1 (n=46) and CMT2 (n=9), were studied. Thirty-two of 46 CMT1 cases (69.6%), all heterozygous but one homozygous for the pVAW409R3a MspI polymorphism, from 12 of 21 families (57.1%) were positive for the CMT1A duplication. In autosomal dominant familial cases (n=30), 26 of 30 cases (86.7%), all heterozygous, from six of seven families (85.7%) were positive for duplication. None of the CMT2 patients, asymptomatic family members or healthy controls were positive for duplication. The CMT1A frequency of duplication in Norwegian CMT1 patients is in general agreement with those reported in other European countries and the present results show that quantitative densitometric PSL imaging is a highly reliable test in diagnosing CMT1A duplication. PMID- 11489281 TI - Human wild presenilin-1 mimics the effect of the mutant presenilin-1 on the processing of Alzheimer's amyloid precursor protein in PC12D cells. AB - Most familial early-onset Alzheimer's disease (FAD) is caused by mutations in the presenilin-1 (PS1) gene. Abeta 42 is derived from amyloid precursor protein (APP) and increased concentrations are widely believed to be a pathological hallmark of abnormal PS function. Thus, the interaction between PS1 and APP is central to the molecular mechanism of AD. To examine the effect of wild-type human PS1 on rat APP metabolism, we made several PC12D cell lines that expressed human wild or mutant PS1, and analyzed the processing of endogenous rat APP and the intracellular gamma-secretase activity. We found the ratio of Abeta 42/Abeta 40 increased in PC12D cells expressing wild-type human PS1. These changes were identical to those found in PC12D cells expressing human PS1 bearing the A260V mutation. These results suggest that APP metabolism is physiologically regulated by the PS1 and that loss of normal PS1 affects gamma-secretase activity. PMID- 11489282 TI - Magnetization transfer and diffusion tensor MR imaging of the optic radiations and calcarine cortex from patients with Leber's hereditary optic neuropathy. AB - Optic nerve abnormalities are easily detectable in patients with Leber's hereditary optic neuropathy (LHON), using magnetic resonance imaging (MRI). However, the presence of structural changes in the optic radiations and calcarine cortex of these patients is still an unresolved issue. In this study, we obtained magnetization transfer (MT) and diffusion tensor (DT) MRI to investigate the integrity of these structures in patients with LHON. Dual echo, MT- and DT-MRI scans of the brain were obtained from 10 men with LHON and 10 healthy sex- and age-matched controls. After image co-registration, we obtained MT ratio (MTR), average diffusivity (D) and fractional anisotropy (FA) maps. MTR, D and FA values for the white matter of the optic radiations and MTR and D values for the calcarine cortex were obtained using a region of interest (ROI) analysis. No macroscopic abnormalities were detected in any of the scans from LHON patients and controls. No statistically significant differences of MTR, D or FA values were found for any of the regions studied in LHON patients and healthy controls. Our results suggest that, in patients with LHON, the optic radiations and the calcarine cortex are spared from structural damage, both at a macroscopic and a microscopic level. PMID- 11489283 TI - Pupillary disturbances in multiple sclerosis: correlation with MRI findings. AB - Autonomic nervous system disturbances such as pupillary abnormalities have rarely been evaluated in multiple sclerosis (MS). However, pupillary impairment is not uncommon in MS and its origin is still unclear. The aim of this study was to investigate pupillary disturbances in MS and to try to correlate pupillary defects with spinal cord and brainstem magnetic resonance imaging (MRI) findings. We prospectively studied 45 MS patients and 30 normal subjects. METHODS: The pupillary contraction latency and the amplitude of contraction were recorded by pupillometry. We also determined afferent and efferent pathway defects by comparing the direct and consensual pupillary reflexes. We evaluated brainstem and spinal cord demyelinating lesions and spinal cord cross-sectional area on MRI. At least one pupillometric parameters were significantly impaired in 60% of patients and in none of the controls. We did not find any correlation between pupillary defect and demyelinating lesions on MRI. The most frequent abnormality was efferent pathway shift and this was correlated with spinal cord atrophy (P<0.02). These results confirm that the autonomic nervous system, and especially pupillary function, is frequently impaired in MS. The parasympathetic system is most commonly affected and this is most likely linked to axonal loss (demonstrated by spinal cord atrophy) rather than to demyelinating lesions. PMID- 11489284 TI - A possible novel isoform of peripheral myelin P0 protein: a target antigen recognized by an autoantibody in a patient with malignant lymphoma and peripheral neuropathy. AB - We tried to characterize a 35-kD antigen recognized by the serum IgG of a patient with malignant lymphoma and peripheral neuropathy. By Western blotting, the serum IgG reacted with the 35-kD antigen in the human, bovine and mouse peripheral nerve (PN) but not with other neural and non-neural tissues. Immunohistochemical analysis showed immunoreactivity of the IgG in the compact myelin of PN. We constructed a human sciatic nerve cDNA library and screened it using IgG of the patient. Three independent clones were obtained. Sequence alignment indicated that the inserts of these clones were homologous to the P0 cDNA, but that all three corresponded to the 3'-untranslational region of the P0 cDNA. To biochemically analyze the 35-kD antigen, myelin fractions of the human and bovine PN were prepared. The 35-kD antigen was purified from the crude myelin fraction by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. When the immunoreactivities of the 35-kD antigen for the IgG of the patient and a monoclonal anti-P0 antibody were compared with those of P0 protein for these antibodies, the 35-kD antigen reacted with both antibodies, but the P0 protein reacted with only the monoclonal anti-P0 antibody. These results suggest that the 35-kD antigen is an isoform of P0 protein. Although it is unlikely that the autoantibody may be the primary cause of neuropathy, because they were also detected in patients with lymphoma without overt neuropathy, they appear to be a modifying factor in the progression of neuropathy. PMID- 11489285 TI - Autosomal dominant Kufs' disease: a cause of early onset dementia. AB - Kufs' disease is the rare, adult-onset form of the neuronal ceroid-lipofuscinoses (NCL). Two clinical Kufs' phenotypes have been described, one featuring generalized tonic-clonic seizures and the other characterized by dementia. Autosomal dominant inheritance of Kufs' disease has been reported for only two families. The genetic and molecular defects underlying Kufs' disease are unknown. We report a third family with apparent autosomal dominant Kufs' disease in a family of English ancestry. Ten individuals (five men, five women) have been affected over five generations. Age of onset typically is in the fourth decade of life and is heralded by seizures. Clinical and neuropsychological assessments in several affected individuals, however, confirm the presence of dementia and follow-up evaluations suggest that dementia is the primary disabling feature of the illness. Motor abnormalities also are frequent. Neuropathological examination (three cases) documents the presence of neuronal lipopigment accumulation consistent with NCL. The combination of dementia and seizures in this and two other reported families with autosomal dominant Kufs' disease suggest that this entity represents a distinctive clinicopathological entity. Dementia is prominent but is almost always associated with generalized seizures and motoric disturbances early in the disease course. Kufs' disease should be considered in the differential diagnosis of early onset, atypical dementia. PMID- 11489286 TI - Cytotoxic T cell activity against the peptide, AYRARALEL, from Yo protein of patients with the HLA A24 or B27 supertype and paraneoplastic cerebellar degeneration. AB - OBJECTIVE: To determine a peptide that reacts with cytotoxic T cells (CTL) of patients with paraneoplastic cerebellar degeneration and anti-Yo antibodies with either HLA A24 or B27 supertype. METHOD: We studied CTL activity of four patients, three were HLA A24-positive and one did not have HLA A24 but had B27 supertype. After an incubation of mononuclear cells with or without peptide and IL-2, CD8-rich fraction was prepared by treatment with Magnetic Cell Sorting system (MACS) twice. CTL activity was calculated by 51Cr release from transfectant, C1RA*2402 as target cells. The peptide-binding assay was examined by flow cytometry. RESULTS: Two of three HLA A 24-positive patients demonstrated CTL activity against the Yo peptide, AYRARALEL. CTL activity was found to be 19.5% and 11.7% at the effector/target (E/T) ratio of 23:1 and 11:1, respectively. A patient who did not have HLA A24 but had A2 and B27 supertype possessed a CTL activity of 19.4% with 15:1 as E/T ratio. The peptide could bind to HLA A*2402 molecules but not to A*0201. CONCLUSIONS: We showed CTL activity in two of three Japanese patients with HLA A24 by using HLA A*2402 transfectant cells as the target. In addition, we identified the first Japanese patient who had B27 supertype, and suggested that the same peptide, AYRARALEL, could be recognized by CTL in this patient. PMID- 11489287 TI - Anti-myelin associated glycoprotein antibodies: variability in patterns of IgM binding to peripheral nerve. AB - We previously found that serums with anti-sulfatide antibodies have several different patterns of binding to neural tissue. In this study, we asked whether serums with anti-myelin associated glycoprotein (MAG) antibodies also have similar variations in patterns of tissue binding. We examined binding to peripheral nerve in 49 serums with IgM anti-MAG antibodies and 13 serums with IgM anti-sulfoglucuronyl paragloboside (SGPG) antibodies but no MAG binding. We correlated patterns of binding with titers of IgM binding to MAG and SGPG measured by ELISA methods. Our results show that IgM in most anti-MAG serums stained areas of non-compact myelin, including the periaxonal and outer myelin membranes and Schmidt-Lanterman incisures. However, other patterns included IgM binding to areas of compact myelin and to non-myelin structures including axons and endoneurial macrophages. IgM in anti-SGPG serums bound to axons or macrophages, but rarely to myelin-related structures. A total of 11/62 (18%) of serums had IgM binding to axons, six with anti-MAG antibodies and five with anti SGPG antibodies. The majority of these serums (73%) had SGPG titers greater than MAG titers when measured by ELISA. We conclude that anti-MAG serums have several different binding patterns to neural tissue, including axonal binding, especially when anti-MAG antibodies cross-react with SGPG. These different binding patterns may relate to the ability of anti-MAG serum IgM to bind both MAG and SGPG or to other molecules with a sulfated glucuronic acid epitope that are present in peripheral nerve. PMID- 11489288 TI - Epilepsy care in six Indian cities: a multicenter study on management and service. AB - BACKGROUND: Epilepsy care in developing countries is lagging behind than in the developed countries. Precise data on delivery of neurological services for epilepsy is essential to optimize the medical services for epilepsy care with limited resources. OBJECTIVE: This study was carried out in order to examine the management practices and utilization of various medical services for epilepsy in different parts of India. METHODOLOGY: University centers with epilepsy clinics, one each from six states of India, had participated in this study. Demographic data, clinical details, and data on epilepsy care were collected simultaneously on standard proforma. RESULTS: Data on 285 patients with epilepsy (generalized epilepsy: 49.1%, localization-related epilepsy: 49.9%, others: 1%) were included. Mean age of onset of epilepsy was 14.8+11.1 years. Mean delay in diagnosis was 1.5+/-4 years. Mean distance from place of residence to the consulting neurologist was 70+/-82 km. Medical consultations before referral to epilepsy center included general practitioners (54.1%) and specialists (43.3%). Very few patients received services from clinical psychologist or social worker. Investigations included, EEG (63.2%), CT Scan (36.2%). MRI brain (8.5%) and video EEG (2.1%) were limited to a few. Nearly 75.5% were on monotherapy. Newer Anti Epileptic Drugs (AEDs) were used only in less than 5% patients. CONCLUSION: The services for epilepsy are urban-based and there is underutilization of services, general practitioners and specialists. Newer AEDs (although expensive) are gradually emerging in Indian market. Facilities for epilepsy surgery, therapeutic drug monitoring and services of clinical psychologist or medical social workers are limited. PMID- 11489289 TI - Magnetization transfer measurements of the hippocampus in the early diagnosis of Alzheimer's disease. AB - We measured magnetization transfer ratios (MTRs) of the hippocampus in 38 patients with Alzheimer's disease (AD), including very mild (Clinical Dementia Rating [CDR] 0.5, n=12), mild (CDR 1, n=14), and moderate stages (CDR 2, n=12), and in 21 healthy elderly control subjects. Medial temporal lobe atrophy was graded subjectively on a five-point scale by two observers blinded to clinical data. Compared with the controls, each of the AD groups, including the very mild group, had significant atrophy of the medial temporal lobe and a decrease in MTRs of the hippocampus. Logistic regression analysis revealed that the overall discrimination rate with MTR measurement and visual analysis of the atrophy was 85% and 73% between the control group and the CDR 0.5 group, 89% and 80% between the control group and the CDR 1 group, and 100% and 91% between the control group and the CDR 2 group, respectively. MTR measurements may provide additional information in detecting structural damage of the hippocampus of AD and be helpful in providing improved diagnosis and early detection of AD. PMID- 11489290 TI - Clinically relevant cognitive impairment after cardiac surgery: a 6-month follow up study. AB - BACKGROUND AND PURPOSE: The majority of studies on neuropsychological complications after cardiac surgery used the raw variation of selective tests scores to define the occurrence of cognitive decline. We prospectively estimated the frequency of cognitive impairment after cardiac surgery, with a particular emphasis on persistent and clinically relevant cognitive decline. Possible baseline and operative predictors were also evaluated. METHODS: An extensive neuropsychological battery was administered to 110 patients (mean age 64.1+/-9.4 years; 70.9% males) undergoing cardiac surgery before and 6 months after the operation. After evaluating the variations in the cognitive performances, two independent neuropsychologists ranked the patients as unchanged-improved, mildly moderately deteriorated, or severely deteriorated, using a global and functionally oriented judgement. The degree of the impairment was determined in relation to its impact on everyday life activities. RESULTS: Ten patients (9.1%) were ranked as severely deteriorated, 22 (20%) as mildly-moderately deteriorated, and 78 (70.9%) as unchanged-improved. Cognitively impaired patients were older (p=0.031), more often females (p=0.005), with a low education level (p=0.013). At multivariate analysis, female gender (odds ratio (OR) 6.14, 95% confidence interval (95% CI) 2.16-17.50), baseline use of beta-blockers (OR 4.55, 95% CI 1.30-15.92), and PaO2 at arrival in intensive care unit (OR for 1 mm Hg increment 1.012, 95% CI 1.004-1.020) were significant predictors of cognitive impairment of any degree. Positive predictors of severe cognitive impairment were history of hypertension (OR 5.33, 95% CI 1.03-27.64) and PaO2 at arrival intensive care unit (OR for 1 mm Hg increment 1.020, 95% CI 1.006-1.035), while education was protective (OR per year of increment 0.53, 95% CI 0.31-0.90). CONCLUSIONS: A considerable proportion of cardiac surgery patients may undergo clinically relevant cognitive impairment. The knowledge of variables influencing cognitive outcome is essential for the adoption of preventive measures. PMID- 11489291 TI - Sweet's syndrome associated with encephalitis. AB - The involvement of the central nervous system (CNS) in Sweet's syndrome (acute febrile neutrophilic dermatosis) is rare. We report a 47-year-old woman who presented with acute encephalitis and was subsequently diagnosed as having Sweet's syndrome. She developed altered consciousness following fever and erythematous skin plaques in the extremities. Cerebrospinal fluid (CSF) examination disclosed neutrophilic pleocytosis without decreased glucose level. Brain magnetic resonance imaging (MRI) showed abnormal signal intensity lesions in the basal ganglia and the hippocampus. Skin biopsy revealed a dense dermal infiltration of neutrophils, which is compatible with Sweet's syndrome. Treatment with acyclovir and antibiotics failed, but the subsequent corticosteroid therapy was effective. Awareness of neurological complication in Sweet's syndrome may avoid unnecessary empiric therapy for meningoencephalitis and will lead to a successful treatment with corticosteroids. PMID- 11489292 TI - Association of the T102C polymorphism of 5-HT2A receptor gene with aura in migraine. AB - OBJECTIVE: To find out the significance of the 5-HT2A receptor gene polymorphism in migraine. STUDY DESIGN: A PCR study in which 61 migraineurs and 44 healthy controls were included. METHODS: The T102C polymorphism of the 5-HT2A receptor gene was studied. The results of the migraineurs and controls were compared. The relationship between the gene polymorphism and aura was also assessed. RESULTS: The representations of the 5-HT2A genotypes were similar in migraineurs and controls (p>0.05) as well as in the male and female migraineurs (p>0.05). The family history of migraine did not associate with 5-HT2A receptor gene polymorphism (p>0.05). There was a significant relationship between the presence of C/C genotype and migraine with aura (p=0.02) while C/T and T/T genotypes were over represented in the patients with migraine without aura (p<0.01). CONCLUSION: The T102C polymorphism of the 5-HT2A receptor gene is not directly related to the increased risk of migraine. The associations between the genotypes of this gene and aura may suggest that 5-HT2A receptor gene polymorphism may be involved in determining the subtypes of or accompanying symptoms in the migraine disease. PMID- 11489293 TI - Sensors for brain mapping. PMID- 11489294 TI - Identification of nuclei associated proteins by 2D-gel electrophoresis and mass spectrometry. AB - In clinical neuroscience as well as in many other clinical disciplines, the completion of the human genome project offers a new possibility to identify and localize the products of the genes, the proteins. Nuclear proteins are synthesized in the cytoplasm and imported into the nucleus by multiple pathways. The mechanisms by which nuclear accumulation of different molecular species occur are unclear but it is apparent that changes in the cellular and molecular events associated with the accumulation of nuclear proteins sometimes precedes transformation of cells into diseased states. The significance of the accumulation and the operation of nuclear proteins remain to be elucidated in detail. Such knowledge will play a key role in the understanding of the regulation of transcription and its disturbances in several of our most devastating diseases. In this paper we present a strategy to identify nuclear associated proteins in small samples by using two-dimensional electrophoresis and mass spectrometry. We have used human blood lymphocytes as a model, but the method should be rather general for any kind of tissue. Twenty two proteins were randomly chosen, and of these 18 proteins were identified by database searching of mass spectrometric data, obtained from in-gel tryptic digests of the spots. Thirteen proteins recently described with nuclear localization and function were identified, and five proteins; calgranulin B, glyceraldehyde-3-phosphate dehydrogenase (G3P2), a TATA-binding protein (ATBP), tubulin beta chain and moesin were also identified as being nuclear associated. The presented data clearly shows of the great role of two-dimensional gel electrophoresis and modern mass spectrometry in the excavation of the protein patterns on the subcellular level, and the ability to use small samples well suited for clinical screening. PMID- 11489295 TI - Effects of recording media composition on the responses of Nafion-coated carbon fiber microelectrodes measured using high-speed chronoamperometry. AB - The present study concerns methodological issues of electrochemical recordings using Nafion-coated 30 microm diameter single carbon fiber microelectrodes for high-speed chronoamperometric measurements of biogenic amines. First, the single carbon fiber microelectrodes were coated with Nafion and dried at 85 vs. 200 degrees C and their recording properties were determined. Second, the effects of shifts in solution pH, ionic strength, changes in recording solution levels of Ca(2+) or Mg(2+) and temperature on the recording characteristics and sensitivity of Nafion-coated high temperature dried (200 degrees C) single carbon fiber microelectrodes for measures of dopamine were studied. These studies showed that the high temperature drying of the Nafion produced a microelectrode with better recording properties: higher selectivity for cations versus anions, increased differences between the reduction and oxidation current ratios for the identification of dopamine versus serotonin, and more rapid response times. In addition, these studies demonstrated that the chronoamperometric recordings were insensitive to small changes in pH and divalent cations such as Ca(2+) or Mg(2+). However, increases in ionic strength decreased the sensitivity of the microelectrodes, while increases in temperature produced increases in the sensitivity of the microelectrodes for biogenic amines. These data support that Nafion-coated high temperature (200 degrees C) dried microelectrodes have enhanced recording properties as compared to microelectrodes, which are coated with Nafion and dried at 85 degrees C. In addition, high-speed chronoamperometric recordings of biogenic amines are not affected by solution changes in divalent cations (Ca(2+) or Mg(2+)). PMID- 11489296 TI - Investigation of the metabolism of substance P in rat striatum by microdialysis sampling and capillary electrophoresis with laser-induced fluorescence detection. AB - The metabolism of substance P (SP) was investigated in rat striatum using in vivo microdialysis. Substance P was perfused for 5 h at 0.2 microl/min, and its metabolism was followed for over 13 h. The resulting samples were derivatized precolumn with naphthalene-2,3-dicarboxaldehyde (NDA)/cyanide, separated and detected by cyclodextrin-modified electrokinetic chromatography with laser induced fluorescence detection (CDMEKC-LIF). Substance P rapidly degraded to form the fragments (3-11), (1-9), (1-4) and, to a lesser extent, (1-7). The metabolites reached steady-state levels 2-3 h after addition of SP. PMID- 11489297 TI - Functional neuroanatomy of the ventral striopallidal GABA pathway. New sites of intervention in the treatment of schizophrenia. AB - Microdialysis was employed to investigate the dopamine, cholecystokinin (CCK) and neurotensin receptor regulation of ventral striopallidal GABA transmission by intra-accumbens perfusion with selective receptor ligands and monitoring local or ipsilateral ventral pallidal GABA release. In the dual probe studies intra accumbens perfusion with the dopamine D1 and D2 receptor agonists SKF28293 and pergolide had no effect on ventral pallidal GABA, while both the D1 and D2 receptor antagonists SCH23390 and raclopride increased ventral pallidal GABA release. In contrast, intra-accumbens CCK decreased ventral pallidal GABA release and this was reversed by local perfusion with the CCK2 receptor antagonist PD134308 but not the CCK1 receptor antagonist L-364,718. In a single probe study intra-accumbens neurotensin increased local GABA release, which was strongly potentiated when the peptidase inhibitor phosphodiepryl 08 was perfused together with neurotensin. In addition, the neurotensin receptor antagonist SR48692 counteracted this phosphodiepryl 08 induced potentiated increased in GABA release. Taken together, these findings indicate that mesolimbic dopamine and CCK exert a respective tonic and phasic inhibition of ventral pallidal GABA release while the antipsychotic activity associated with D1 and D2 receptor antagonists may be explained by their ability to increase ventral striopallidal GABA transmission. Furthermore, the findings suggest that CCK2 receptor antagonists and neurotensin endopeptidase inhibitors may be useful antipsychotics. PMID- 11489298 TI - Development of silicon microstructures and thin-film MALDI target plates for automated proteomics sample identifications. AB - Here we report on the development of a proteomic platform utilizing a piezoelectric flow-through dispensing unit made from silicon microstructures. The use of a novel surface coating, where matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI MS) targets were uniformly precoated with a thin film of matrix/nitrocellulose, made the sample preparation straightforward and enabled the enrichment and analysis of proteins at low levels in proteomics samples. We demonstrate this by analyzing excised spots in a biological sample originating from a human fetal fibroblast cell line that was subjected to 2D gel electrophoresis. Furthermore, a sample deposition rate below 30 Hz results in an increased analyte density on the dispensed sample spot, rendering signal amplification. In general, the sensitivity for proteins and peptides can be enhanced 10-50 times compared to traditional MALDI sample preparation techniques. PMID- 11489299 TI - Effect of intracerebral 6-nitronoradrenaline, an endogenous catechol-O methyltransferase (COMT) inhibitor, on striatal dopamine metabolism in anaesthetised rats. AB - 6-Nitronoradrenaline, a bioactive compound recently identified in the brain, is known to inhibit catechol-O-methyltransferase. To study its effect on dopamine metabolism, it was administered into rat striatum via a microdialysis probe. Other nitrated catechols (6-nitrodopamine, 6-nitro-DOPAC and 5-nitro-HVA) were studied for comparison. Tolcapone, a selective catechol-O-methyltransferase inhibitor, was used as a positive reference compound. Both 6-nitronoradrenaline and tolcapone increased striatal extracellular dopamine levels during the perfusion (at 100 microM concentration but not at 10 microM) and decreased the efflux of homovanillic acid. Tolcapone, but not other nitrated catechols, increased 3,4-dihydroxyphenylacetic acid efflux. None of the compounds inhibited MAO-B activity at 100 microM or lower. At 1 mM, 6-nitrodopamine inhibited MAO-B by 60%. Compared to tolcapone, other nitrated catechols were very weak COMT inhibitors in vitro. Neither tolcapone nor 6-nitronoradrenaline modified the metabolism of L-dopa which was given peripherally. In binding studies, both 6 nitronoradrenaline and other nitrocatechols failed to affect the dopamine transporter even at high micromolar concentrations. In conclusion, exogenous 6 nitronoradrenaline can act as a COMT inhibitor in the striatum and elevate striatal dopamine levels without inhibiting dopamine reuptake. Whether endogenous 6-nitronoradrenaline can be formed also in vivo in the striatum and act as a regulator of dopaminergic tone remains to be determined. PMID- 11489300 TI - A method to evaluate the diffusion rate of drugs from a microdialysis probe through brain tissue. AB - For interpretation of microdialysis experiments in which compounds are applied retrodialysis, it is important to have information about the migration rate of the infused compounds. Here we describe a dual-probe microdialysis method that can be used to evaluate the penetration rate of the infused drug. The basic idea is that not the drug itself is assayed, but that its pharmacological effect is recorded by a second probe positioned at a fixed distance (1 mm) of the infusion probe. Using this approach several compounds, each known to induce specific changes in the extracellular levels of dopamine, were infused into the striatum of the rat. The results indicate that the penetration rate of the pharmacological effect of infused compounds differed widely. No effects were seen at the second probe when high potassium chloride was infused. Apparently dopamine was not able to migrate into brain tissue over a distance of 1 mm. Low penetration rates were observed for the dopamine antagonist sulpiride, the dopamine agonist LY17155, and for amphetamine and nomifensine. A very high penetration rate was observed in case of tetrodotoxin. The fast effects of TTX could also be explained by remote inhibition of neurons passing along the infusion probe. The present study showed that most of the compounds have rather slow infusion rates, indicating that relatively high infusion concentrations are needed (1-10 mM) to reach substantial brain concentrations at a distance of 1 mm from the infusion probe. PMID- 11489302 TI - Microdialysis in freely moving mice: determination of acetylcholine, serotonin and noradrenaline release in galanin transgenic mice. AB - In the present study, we describe micro-surgical methods for simultaneous implantation of a microdialysis probe and an intraventricular injection cannula via their respective guide cannulas into the mouse brain. Basal and stimulated release of acetylcholine (ACh), serotonin (5-HT) and noradrenaline (NA) was determined in the ventral hippocampus of freely moving mice. NA and 5-HT were determined in one run by a newly developed HPLC method based on precolumn derivatization with benzylamine and fluorescence detection. The mice with a loss of-function mutation of the galanin gene (KO) and the mice that over-expressed galanin (OE) were studied. No significant differences in basal, potassium stimulated or scopolamine-induced extracellular ACh levels were observed in 4 month-old wild-type (WT) and KO mice. In the aged, 10-month-old animals, the basal extracellular ACh levels were significantly reduced in both WT and KO groups. Galanin (1 nmol i.c.v.) caused a significant reduction of basal extracellular NA by about 40% in both WT and galanin OE mice, however, in the latter group the effect was delayed by almost 2 h. A 10-min forced swimming stress caused a higher increase in release of NA and 5-HT in the OE group than in the corresponding WT mice. Finally, venlafaxin (10 mg/kg i.p.) increased extracellular NA to 400% of the control values in the CBA mice, but only to 250% in the C57BL mice. It is concluded that galanin may play an important role in the cholinergic mechanisms underlying cognitive disorders. Furthermore, modulation by galanin and by behavioral activation, of NA and 5-HT neurotransmission in galanin over-expressing mice indicates its possible role in the aetiology of mood disorders. PMID- 11489301 TI - Can voltammetry measure nitrogen monoxide (NO) and/or nitrites? AB - Recently, voltammetry with carbon fibre electrodes (CFE) has been implemented for real time measurement of nitrogen monoxide (NO) indicating that it is oxidised at the potential value of nitrites, approximately +700 mV. In contrast, here we show that modified CFE can monitor NO at oxidation potentials different than that of nitrites, i.e. +550 mV. Indeed, at +550 mV a significant increase of amperometric current levels was obtained when NO but not nitrites, were added to a phosphate buffer saline solution (PBS). Differential pulse voltammetry (DPV) supports these findings as two oxidation peaks were obtained when examining air preserved NO; peak 1 at +550 mV and peak 2 at +700 mV, respectively. In contrast, only peak 2 was monitored when nitrites or a solution of NO oxidised in air was added to PBS. Biological support to these in vitro data comes from the observation that the relaxation of an adrenaline-contracted aortic ring produced via addition of NO is concomitant with peak 1 at +550 mV. The relaxation is almost completed before the appearance of peak 2 at +700 mV. Furthermore, in vivo experiments performed in the striatum of rats show that the amperometric signal monitored at +550 mV is responsive to glutamatergic stimulation or inhibition of NO synthase. PMID- 11489303 TI - Effect of temperature on the uptake of waterborne strontium in the common carp, Cyprinus carpio (L.). AB - The effect of temperature on the uptake kinetics of strontium (Sr) in the common carp (Cyprinus carpio) was studied in vivo, exposing pre-acclimated fish to a wide range of Sr concentrations in water (Sr(total)=0.2-10,000 microM; Ca(total)=348 microM) at 10, 20, 25 and 30 degrees C. Sr uptake rates were determined in the whole body, gills and blood of the fish after an exposure period of 3 h and were analyzed as a function of the free-ion activity of Sr and Ca in water. The uptake of Sr(2+) by the whole body, gills and blood increased with temperature and showed saturation kinetics with the increase of Sr(2+) activity. Analyzing the observed uptake rates with a Michaelis-Menten type model showed that the kinetic parameters (J(max), K(m) and K(i)) for both Sr(2+) and its analogue Ca(2+) are temperature dependent. Thermodynamic analysis of the temperature effects indicates that the Arrhenius activation energies (E(a)) required for Sr(2+) uptake (91.9 kJ mol(-1)) and Ca(2+) uptake (105.9 kJ mol(-1)) in the whole body of carp were constant over the temperature range 10-25 degrees C and showed a break in the Arrhenius plots above this temperature. The Arrhenius plot for the Sr(2+) uptake in blood was similar to that for the whole body uptake with an E(a) of 98.1 kJ mol(-1). However, the E(a) for Sr(2+) uptake in gills was much smaller and constant (58.1 kJ mol(-1)) over the temperature range of 10-30 degrees C. For a temperature change from 10 to 25 degrees C, the Q(10) for Sr(2+) uptake in whole fish, gills and blood were 3.71, 2.29 and 4.05, respectively. Compared with Ca(2+) uptake, Sr(2+) uptake appears to require a lower activation energy for transport across the solution body interface in carp. The similar pattern of Arrhenius plots and magnitude of activation energies for Sr(2+) uptake both in blood and whole fish suggest that the uptake into the blood across the basolateral membrane is the rate-limiting energy barrier and hence dictates the overall uptake rate of Sr(2+) in whole fish. PMID- 11489304 TI - Acute and chronic physiological effects of silver exposure in three marine teleosts. AB - This study evaluated the physiological effects of waterborne silver (added as AgNO(3)) on seawater fish, using acute (48-72 h) high level exposures (250-650 microg/l Ag) on tidepool sculpins (Oligocottus maculosus), and chronic (up to 21 day) low level exposures (1.5-50 microg/l Ag) on tidepool sculpins, plainfin midshipmen (Porichthys notatus), and rainbow trout (Oncorhynchus mykiss). Sculpins were tested at different salinities. Acclimation to lower salinity (18 vs 30 ppt) led to altered physiology, with higher ammonia excretion (J(Amm)), lower oxygen consumption, and lower branchial and intestinal Na(+)/K(+)-ATPase activities, but no difference in drinking rate. Short-term exposure to high silver levels tended to stimulate M(O(2)), J(Amm), and drinking rate. However, long-term exposure to low levels of silver depressed both J(Amm) and M(O(2)), and also led to decreased drinking rates. Both inhibition and stimulation of Na(+)/K(+)-ATPase activity occurred, dependent upon length and concentration of exposure, salinity (18 vs 30 ppt), tissue (gill vs intestine), and fish species (sculpin vs midshipmen vs rainbow trout). While the effects were variable, due to differing balances between inhibitory and compensatory responses, chronic silver exposure significantly altered Na(+)/K(+)-ATPase activity levels in almost all tests. In total, these findings reinforce the view that intestinal osmoregulatory function (drinking, Na(+)/K(+)-ATPase activity) is an important site of toxic impact for waterborne silver, that gill Na(+)/K(+)-ATPase activity is also a site of impact, and that chronic exposures at silver concentrations (1.5, 14.5 microg/l Ag) close to current or proposed water quality guidelines (albeit much higher than normal environmental levels), exert a variety of sublethal effects on marine teleosts. PMID- 11489305 TI - Influence of o'p-DDD on the physiological response to stress in Arctic charr (Salvelinus alpinus). AB - Various toxicants have previously been held responsible for an impaired capacity of fish from polluted environments to elevate their cortisol levels in response to stress. In the present study we investigated the responses to stress in o'p DDD [2-(chlorophenyl)-2-(4-chlorphenyl)-1,1-dichloroethane] exposed (given a single, oral dose of 75 mg o'p-DDD/kg fish) and unexposed Arctic charr. After o'p DDD administration fish were left undisturbed and without being fed for 28 days, when they were subjected to an acute handling stress. At 1, 3, 7 and 23 h following stress, primary (ACTH and cortisol secretion) and secondary (plasma Cl levels and energy mobilisation) components of the stress response were monitored. As the nutritional state of wild fish may influence this potential biomarker response, the fish had been subjected to a restricted feed ration prior to o'p DDD administration in order to obtain marked within-group variations in condition factor. No effects of o'p-DDD were observed on post-stress hormone secretion (i.e. peak post-stress plasma ACTH and cortisol levels), nor on plasma chloride levels. However, other results obtained provided evidence for a metabolic depression by o'p-DDD, witnessed by consistently lower plasma glucose levels before and after stress in these contaminated fish. This may be related to the finding that during the 30-day period between o'p-DDD administration and stress treatment, toxicant treated fish lost less weight in comparison to their sham treated counterparts. Nutritional state did not appear to influence the performance of the charr in the present experiment, as correlations between the parameters measured and condition factor or lipid contents on an individual basis in all cases turned out non significant. Overall, the results contrast with those of previous in vivo and in vitro studies on fish, which concluded that comparable headkidney o'p-DDD levels impaired interrenal steroidogenesis. Although we conclude that the effects of o'p-DDD on Arctic charr metabolism were not associated with the stress response, we propose that they may well interfere with the animals' ability to cope with stress in the long term, or may compromise other physiological processes, such as smoltification. Finally, the high level of integration of components involved in the stress response complicates the identification of single stress-sensitive indices as biomarkers applicable in environmental monitoring programmes. PMID- 11489306 TI - Expression of P-glycoprotein in the gills of oysters, Crassostrea virginica: seasonal and pollutant related effects. AB - The expression of p-glycoprotein (p-gp) in aquatic organisms has been proposed as a biomarker of pollution exposure. Previous research has provided evidence that p gp is inducible by organic xenobiotics and that p-gp is overexpressed in mussels from degraded areas. However, seasonal changes in expression at polluted sites has not been described previously. The purposes of these studies were to evaluate the expression of p-gp in polluted and unpolluted sites in oysters (Crassostrea virginica) and to determine if there were seasonal differences. P-gp expression and total protein concentrations were measured seasonally in the gills of southeastern oysters, Crassostrea virginica, at degraded and undegraded sites in Charleston Harbor, South Carolina, USA. At all sites, p-gp expression was generally higher during the warmer months and lower in the colder months. Polluted sites tended to show a decrease in p-gp expression in June and September, suggesting that p-gp inhibition may have occurred. P-gp expression was not significantly related to specific classes of sediment contaminants or to overall sediment contaminant loading. Total gill protein concentrations at all sites were lower during the warmer months and higher during the colder months. In general, all sites tended to show decreased total gill protein concentrations when compared to the control site. Total protein concentrations were significantly related to water temperature, specific classes of sediment contaminants and overall sediment contaminant loads, but there was no relationship with salinity. PMID- 11489307 TI - Interaction of cadmium toxicity in embryos and larvae of zebrafish (Danio rerio) with calcium and humic substances. AB - The influence of humic substances (HS) and calcium (Ca) on cadmium (Cd) toxicity was determined using zebrafish (Danio rerio). Embryo and larvae of the zebrafish were exposed to various Cd concentrations (1.8; 2.8; 4.2; 6.2; 9.3 mg/l Cd) for 144 h. Combinations of low (0.2 mmol/l) and high (2 mmol/l) Ca, +HS (5 mg/l C) or -HS were used during Cd exposure. The toxicity of Cd was affected by (1) exposure concentration; (2) exposure time; (3) presence of HS; and (4) the Ca concentration. The results show that Ca and HS protect against Cd toxicity in zebrafish embryos. The best protection was in the high Ca-HS group, followed by high Ca+HS group and low Ca+HS group. The survival in the low Ca-HS group was the worse. Survival in the high Ca-HS group and the high Ca+HS group was similar with the exception of the highest Cd concentration (9.3 mg/l) where the survival of the high Ca+HS group was less than in the high Ca-HS group. The exposure system was modelled using a chemical equilibrium program (MINEQL+) to determine if the likely mechanism causing the anomalous result in the highest Cd concentration. The equilibrium model cannot explain these results, which suggests that this effect has a kinetic basis, such as time needed for Cd to displace Ca already bound by HS. PMID- 11489308 TI - Is hepatic cytochrome P4501A1 expression predictive of hepatic burdens of dioxins, furans, and PCBs in Atlantic tomcod from the Hudson River estuary? AB - Hepatic cytochrome P4501A1 (CYP1A1) expression in fishes is frequently used to evaluate bioavailable aromatic hydrocarbon contamination of aquatic ecosystems. In controlled laboratory experiments, CYP1A1 expression in naive fishes is usually dose-responsive to aromatic hydrocarbons and in field studies levels of gene expression in natural populations often correspond with known levels of sediment-borne contaminants. We quantified CYP1A1 mRNA levels in juvenile Atlantic tomcod Microgadus tomcod from 42 sites in the Hudson River estuary to evaluate the correspondence between hepatic CYP1A1 expression and hepatic concentrations of persistent halogenated aromatic hydrocarbons and to determine the utility of CYP1A1 expression as a biomarker in evaluating the microgeographic distribution of bioavailable contaminants within a large aquatic ecosystem. We found significant spatial heterogeneity in CYP1A1 mRNA levels among collection sites with levels of gene expression differing in some cases by 23-34 folds. CYP1A1 mRNA expression was highest in tomcod from the Newark Bay complex and lowest in tomcod from the most upriver collection sites in the main stem of the Hudson River. Although levels of PCDDs, PCDFs, and PCBs expressed as TCDD TEQs and CYP1A1 mRNA were highest in tomcod from the Newark Bay complex, there was no relationship between hepatic halogenated aromatic hydrocarbon levels and hepatic CYP1A1 mRNA in tomcod from sites in the main stem of the Hudson River. These results suggest that levels of CYP1A1 expression in fish from sites highly polluted with mixtures of halogenated aromatic hydrocarbons and other xenobiotics may not always be reflective of levels of bioavailable aromatic hydrocarbon contaminants. Based on these results and earlier controlled laboratory experiments, we hypothesize that elevated levels of CYP1A1 expression in tomcod from the Hudson River may be due primarily to PAHs or other contaminants not measured in this study. PMID- 11489309 TI - Histological, growth and 7-ethoxyresorufin O-deethylase (EROD) activity responses of greenback flounder Rhombosolea tapirina to contaminated marine sediment and diet. AB - Pathological abnormalities and mixed function oxygenase (MFO) enzyme changes are frequently used as indicators of anthropogenic contaminant exposure and effect. However, there is a paucity of research investigating the effects of contaminated sediment on native Australian benthic teleosts. As part of an ecotoxicological assessment of contaminated marine sediments in northern Tasmania, CYP1A induction, histological and growth response of the greenback flounder, Rhombosolea tapirina, exposed to contaminated marine sediments were examined. Hatchery reared flounder were exposed to reference sediment, contaminated sediment or contaminated sediment and diet for 6 weeks. CYP1A induction, using the ethoxyresorufin-O-deethylase (EROD) assay, and the histological and growth response in the flounder were examined on cessation of the exposure trial. Significant differences were found between treatments in histological, growth and EROD response. Exposure to contaminated sediment and diet elicited a multi-organ histological response: principally partial and total epidermal erosion and multifocal necrosis of the liver. The prevalence of total epidermal erosion was greatest with exposure to disturbed contaminated sediment (66.65+/-16.65%). The prevalence of multifocal necrosis of the liver was greatest with exposure to contaminated sediment and diet (66.65+/-16.65%). Growth reduction, measured as percentage growth inhibition, was evident in flounder exposed to contaminated sediment and diet (18.2+/-11.99%). Additionally, exposure to contaminated sediment and diet elicited elevated induction of the EROD liver detoxification enzyme (139.65+/-24.22 pmol/min/mg protein) compared to exposure to contaminated sediment and non-contaminated diet (6.25+/-0.81 pmol/min/mg) indicating the presence and potential bioavailability of xenobiotics via food. Further, more inhibited growth and histological alteration associated with exposure to contaminated sediment and diet suggest contaminants in Deceitful Cove sediment are cytotoxic. PMID- 11489310 TI - Molecular cloning and characterization of a metal responsive Chironomus tentans alpha-tubulin cDNA. AB - Metal pollution of aquatic ecosystems is a problem of economic and health importance. Sensitive molecular biomarkers of metal exposure are sorely needed. We have isolated a cDNA from the midge Chironomus tentans that is transcribed in all organs and developmental stages. The cDNA encodes a protein, designated Chironomus tentans alpha-tubulin 1 (CTTUB1), which has significant similarities with invertebrate and vertebrate alpha-tubulins. CTTUB1 is abundantly transcribed in embryos and to a lesser extent in adults and larvae. CTTUB1 RNA and protein abundances are increased in larvae exposed to copper or cadmium. The pattern of cellular distribution of CTTUB1 protein in the midgut epithelial cells was radically affected by cadmium. In the midgut cells of unexposed larvae, CTTUB1 was found evenly distributed throughout the cytoplasm, while in cadmium-exposed larvae, CTTUB1 was mostly concentrated along the basolateral plasma membrane. A mechanism for the regulation of alpha-tubulin synthesis by cadmium is proposed. This is the first report on the isolation of a metal responsive gene from a neartic aquatic insect. PMID- 11489311 TI - Sensitivity of the spiny dogfish (Squalus acanthias) to waterborne silver exposure. AB - The physiological effects of waterborne silver exposure (added as AgNO(3)) on spiny dogfish, Squalus acanthias, were evaluated at 30, 200 and 685 microg silver per l in 30 per thousand seawater. These concentrations cover the toxic range observed for freshwater teleosts, where silver is extremely toxic, to seawater teleosts which tolerate higher silver concentrations. However, these levels are considerably higher than those that occur in the normal environment. At 685 microg l(-1), dogfish died within 24 h. Causes of death were respiratory as well as osmoregulatory failure. Arterial P(a)O(2) rapidly declined below 20 Torr, and blood acidosis (both respiratory and metabolic) occurred. Urea excretion increased dramatically and plasma urea dropped from 340 to 225 mM. There were pronounced increases in plasma Na(+), Cl(-), and Mg(2+), indicative of ionoregulatory failure due to increased diffusive permeability as well as inhibited NaCl excretion. At 200 microg l(-1), fish died between 24 and 72 h of silver exposure. The same physiological events occurred with a small time delay. At 30 microg l(-1), effects were much less severe, although slight mortality (12.5%) still occurred. Respiratory alkalosis occurred, together with moderate elevations in plasma Na(+) and Cl(-) levels. Silver accumulated to the highest concentrations on gills, with only low levels in the intestine, in accord with the virtual absence of drinking. Na(+)/K(+)-ATP-ase activities of gill and rectal gland tissue were impaired at the highest silver concentration. Normal gill function was impaired due to swelling and fusion of lamellae, lamellar aneurism and lifting of the lamellar epithelium. Our results clearly indicate that this elasmobranch is much more sensitive (about 10-fold) to silver than marine teleosts, with silver's toxic action exerted on the gill rather than on the intestine, in contrast to the latter. PMID- 11489312 TI - Nanoparticles of a polyelectrolyte-fatty acid complex: carriers for Q10 and triiodothyronine. AB - Poly(ethylene imine) (PEI) was used for the complexation of dodecanoic acid (C12) resulting in a poly(ethylene imine) dodecanoate complex (PEI-C12) with a lamellar nanostructure and a repeat unit of 2.9 nm. PEI-C12 was doped with coenzyme Q10 and the hormone triiodothyronine as typical hydrophobic and pharmacological active compounds, respectively. The PEI-C12 acts as a guest matrix that dissolves both molecules up to weight uptakes of about 20% (w/w) and 15% (w/w), respectively, both without crystallization. Agglomerate-free dispersions of core shell type nanoparticles were developed. Ratios of PEI to C12 of 2:1 or higher were found to be suitable for this purpose. The particles exhibit hydrodynamic diameters in the range of 80-150 nm, which depend on the preparation conditions. Each particle consists of a relatively compact core surrounded by a diffuse corona. PEI-C12 forms the core, while non-complexed PEI acts as a cationic-active dispersing agent. It was found that the nanoparticles show high zeta potentials (approximately +40 mV) and are stable in NaCl solutions at concentrations of up to 0.3 mol x l(-1). The stabilization of the nanoparticles results from a combination of ionic and steric contributions. A variation of the pH value was used to activate the dissolution of the particles. The PEI-C12 nanoparticles may have potential as carriers for hydrophobic drugs. PMID- 11489313 TI - Drug-polyionic block copolymer interactions for micelle formation: physicochemical characterisation. AB - While covalent attachment of small drug molecules to AB copolymers for the formation of polymeric micelles for drug delivery has been investigated, few studies have focused on non-covalent interactions. The aim of this study was therefore to explore the potential of non-covalent interactions between an AB copolymer, Poly(aspartic acid)-poly(ethylene glycol) (Pasp-PEG), with anionic pendant groups and diminazene aceturate, a small molecular weight cationic drug. Micelles were prepared by mixing solutions of Pasp-PEG and diminazene in 25 mM Tris-HCl buffer. At all Pasp-PEG concentrations studied, the micelles appeared to be water soluble with a unimodal size distribution and ranged in size from approximately 22 to 60 nm. The polyionic micelles also displayed similar and small absolute zeta potential values at various drug:monomer molar ratios which confirmed stabilisation by the PEG corona. The scattering intensity was maximal and remained unchanged, while particle size increased slightly at pH range from 3.4 to 7.2. At this pH range both the polymer and drug would be ionised and ionic interactions possible to drive micellar formation. An increase in size and scattering intensity with addition of NaCl to the micelles was attributed to dehydration of the PEG corona which may have led to aggregation of the micelles. The absence of micellar dissociation upon addition of salt was attributed to the dominance of hydrogen bonding between Pasp and diminazene aceturate, as assessed by isothermal titration microcalorimetry. Morphological evaluation of these constructs showed them to be discrete and fairly uniform in size and shape. This study was therefore successful in confirming the potential of non-covalent interactions using an AB copolymer to form polyionic micelles for drug delivery. PMID- 11489314 TI - Preparation and evaluation of poly(L-lactic acid) microspheres containing rhEGF for chronic gastric ulcer healing. AB - Biodegradable microspheres containing recombinant human epidermal growth factor (rhEGF) were prepared using poly(L-lactic acid) by a solvent evaporation method based on multiple w/o/w emulsion. Encapsulation efficiency and initial release were influenced by the amount of polymer, inner water phase volume and osmotic pressure difference between inner water phase and outer water phase. The effect of osmotic pressure difference between inner water phase and outer water phase in w/o/w emulsion on particle size, porosity and in vitro release of rhEGF from microspheres were also studied. Microspheres prepared with the optimized osmotic pressure, polymer amount and inner water volume produced 21% initial release on the first day with 92% encapsulation efficiency. The blood concentration of rhEGF was maintained at constant levels for 9-11 days after a single subcutaneous (s.c.) administration of rhEGF microspheres. The gastric ulcer healing effect of a single s.c. administration of rhEGF microspheres was increased 1.44-fold compared with twice a day s.c. administration of rhEGF saline solution after 11 days. The enhanced curative ratio of rhEGF loaded microspheres may be due to the optimized osmotic pressure, high encapsulation efficiency and sustained release pattern. PMID- 11489315 TI - Sustained release ketoprofen microparticles with ethylcellulose and carboxymethylethylcellulose. AB - Microparticulate systems for sustained release of ketoprofen were prepared and evaluated by monitoring drug release in the JP XIII second fluid, pH 6.8. All the microparticulate dosage forms were prepared using ketoprofen in the form of calcium salt (KP-Ca). Simple ethylcellulose microparticles of KP-Ca (EC-MP) exhibited the fairly rapid release in the first phase with slower release in the late period. Most of the drug was released from EC-MP showing high drug content. For polymer-coated microparticles of ketoprofen, Eudragit microparticles of KP-Ca (ER-MP) were first prepared, and then coated with ethylcellulose or with a mixture of carboxymethylethylcellulose and ethylcellulose to produce ethylcellulose-coated (EC-coat) and the mixture-coated microparticles (CMEC/EC coat), respectively. Some polymer-coated microparticles showed drug release at nearly zero-order rate. Especially, CMEC/EC-coat prepared at a CMEC:EC ratio of 1:1 (w/w), named formation I, could supply the drug constantly and efficiently for about half a day except for an initial rapid release. When formation I was administered intraduodenally to rats, the plasma concentration of ketoprofen could be maintained at a nearly constant level. Kinetic analysis demonstrated that formation I showed a nearly zero-order release rate in vivo consistent with that observed in vitro. PMID- 11489316 TI - Interrelation of permeation and penetration parameters obtained from in vitro experiments with human skin and skin equivalents. AB - In a comparative study, two different in vitro cutaneous test systems were examined: (1) The Franz diffusion cell (FD-C), a test system to study drug permeation through the skin and to obtain data like steady state flux and lag time as well as permeability and diffusion coefficients. (2) The Saarbruecken penetration model (SB-M), a test system to investigate drug penetration into different skin layers and after varying incubation times to acquire values about the quasi steady state drug amounts in the stratum corneum (SC). Three drug concentrations (0.9, 0.45 and 0.225%) of a lipophilic model drug preparation, flufenamic acid in wool alcohols ointment, were applied on the skin's surface using 'infinite dose' conditions. Trypsin-isolated SC, heat-separated epidermis, full-thickness skin and reconstructed human skin (RHS) served as skin membranes in the FD-C, while the SB-M experiments were only carried out using full thickness skin. Increasing steady state flux data and m(ss) values (steady state drug amount in the SC) were detectable after the application of rising drug amounts. Concerning the permeability of the used skin membranes in establishing barrier properties, the following rank order was observed: RHS>SC> or =epidermis>full skin. The flux data of the FD-C experiments for isolated SC, separated epidermis and RHS were linearly related with the m(ss) values of the SB M investigations, allowing a direct comparison of permeation with penetration parameters. Concerning the drug amount in the SC, previous investigations succeeded in the establishment of an in vivo/in vitro correlation. Based on the results presented here, the prediction of drug amounts present in the SC after different incubation times in vivo is now possible after penetration as well as permeation experiments using the lipophilic model drug preparation, flufenamic acid in wool alcohols ointment. PMID- 11489317 TI - Application of the minimal trauma tissue biopsy to transdermal clinical pharmacokinetic studies. AB - Although the transdermal administration of drugs has gained considerable importance, reliable methods for the quantitative assessment of transdermal drug penetration are scarce. The aim of the present study was therefore to evaluate the scopes and limits of the minimal trauma tissue biopsy (MTTB) technique for the in vivo characterization of the transdermal penetration process and the assessment of dermal drug kinetics in humans following topical drug application. Nicotine TTS (21 mg/24 h) was administered transdermally to 13 healthy volunteers. Repeated minimally invasive dermal and subdermal tissue biopsies were obtained at defined time points from defined skin layers directly underlying the TTS. The position of the biopsy needle and depth of biopsate were determined by 2D ultrasound scanning. The biopsy procedure was well tolerated by all volunteers and up to six biopsies within a period of 10 h were easily accepted. Dermal pharmacokinetic profiles for nicotine were obtained in all experiments and corresponded well to the values measured in previous studies on transdermal nicotine penetration. Mean area under the nicotine concentration time curve (AUC) in subepidermal layers underneath the application site of the TTS was 70.0+/-55.1 microg/g per h. There was a correlation between the depth of biopsy sampling and dermal nicotine concentrations at steady state (r=0.7). The MTTB is a suitable, well tolerated technique for the detection of transdermally applied compounds in defined subepidermal tissue layers and could therefore become a valuable tool in the development and assessment of transdermal dosage forms. PMID- 11489318 TI - Preparation, characterization and in vivo evaluation of 120-day poly(D,L-lactide) leuprolide microspheres. AB - A 120-day poly(D,L-lactide) (PLA) microsphere delivery system for a luteinizing hormone-releasing hormone (LHRH) analogue, leuprolide, was prepared and evaluated. Leuprolide microspheres were prepared with PLA (m.w. 11000 Da) by a dispersion/solvent extraction-evaporation method and characterized for drug load by HPLC, particle size by laser diffractometry and surface morphology by scanning electron microscopy. In vitro peptide release and polymer degradation were studied using a modified dialysis method. Serum peptide and testosterone levels were analyzed after subcutaneous administration using a rat model. Spherical microspheres with a mean diameter of 52 microm containing 13.4% peptide released 10% of the peptide within 24 h, followed by a linear release for 150 days. Serum leuprolide levels increased immediately after administration of the microspheres to 45.6 ng/ml, but then fell to 4.3 ng/ml at 15 days and approximately 2.0 ng/ml at 30 days where they remained for 120 days. The testosterone levels increased initially to 15 ng/ml and then decreased to below 0.5 ng/ml by day 4 where they remained for 120 days. In conclusion, a 120-day microsphere formulation of leuprolide was developed with excellent controlled peptide release characteristics and in vivo efficacy. PMID- 11489319 TI - Gamma irradiation effects on stability of poly(lactide-co-glycolide) microspheres containing clonazepam. AB - This work was aimed at evaluating the effects of gamma irradiation on the stability of microspheres made of a poly(lactide-co-glycolide) copolymer (PLGA) and loaded with 15% w/w of clonazepam (CLO). The influence of CLO on PLGA radiolysis mechanisms and the identification of possible irradiation markers were also investigated. Microspheres were prepared by means of a spray-drying method. gamma Irradiation was carried out either under vacuum or in air, at a dose of 25 kGy, by using a 60Co source. The stability of CLO loaded microspheres was evaluated over a 6-month period on the basis of drug content and dissolution profile. Radiolysis mechanisms were investigated by using electronic paramagnetic resonance (EPR) analysis. The microspheres irradiated under vacuum were stable over the considered period of time. After irradiation in air, CLO release rate increased by approximately 10%, and did not change further in the following period of storage. The EPR analysis showed some radicals arising from both the polymeric matrix and the active ingredient. Polymer/CLO spin transfer reactions suggest that CLO had a radio-stabilising effect on the polymeric matrix. In the loaded microspheres, the intensity in time of the CLO radical signal is sufficient for its possible use as irradiation marker. PMID- 11489320 TI - Chemically modified polyacrylamide-g-guar gum-based crosslinked anionic microgels as pH-sensitive drug delivery systems: preparation and characterization. AB - New spherically shaped cross-linked hydrogels of polyacrylamide-grafted guar gum were prepared by the emulsification method. These were selectively derivatized by saponification of the -CONH2 group to the -COOH group. The derived microgels were characterized by FTIR and elemental analyses. The derivatized microgels were responsive to pH and ionic strength of the external medium. The swelling of microgels increased when the pH of the medium changed from acidic to alkaline. Transport parameters, viz., solvent front velocity and diffusion coefficients were calculated from a measurement of the dimensional response of the microgels under variable pH conditions. The variation in pH changed the transport mechanism from Case II (in 0.1 N HCl) to non-Fickian (in pH 7.4 buffer), and these processes are relaxation-controlled. Ionic strength exerted a profound influence on the swelling of the microgels. Swelling was reversible and pulsatile with the changing environmental conditions. The pH-sensitive microgels were loaded with diltiazem hydrochloride and nifedipine (both antihypertensive drugs) and their release studies were performed in both the simulated gastric and intestinal pH conditions. The release was relatively quicker in pH 7.4 buffer than observed in 0.1 N HCl; the release followed non-Fickian transport in almost all the cases. PMID- 11489321 TI - Microscopic localization of PEG-liposomes in a rat model of focal infection. AB - In the present study the microscopic localization of polyethylene glycol (PEG) liposomes in infected tissues was studied with both light microscopy (LM) and transmission electron microscopy (TEM) in rats with focal intramuscular Staphylococcus aureus infection. PEG-liposomes containing colloidal gold were prepared and injected intravenously in rats with focal S. aureus infection and tissues were dissected at 24 h post injection. Sections were cut and liposomes were visualized for microscopic evaluation using silver enhancement. Uptake of PEG-liposomes was visualized by both scintigraphy and LM in the abscess, liver and spleen. In the infected area, the liposomes were mainly found in the vicinity of blood vessels. TEM showed that the liposomes were localized in the macrophages and to a lesser extent in endothelial cells in the infectious tissue. In the liver, the liposomes appeared mainly localized in Kupffer cells. In the spleen, uptake was only seen in cells of the red pulp and in cells around the central arteries. Our microscopic observations indicate that uptake and retention of PEG liposomes in the infectious focus is a result of enhanced extravasation due to increased vascular permeability and subsequent phagocytosis of PEG-liposomes by macrophages in the infected tissue. PMID- 11489322 TI - Trypsin inhibition, calcium and zinc ion binding of starch-g-poly(acrylic acid) copolymers and starch/poly(acrylic acid) mixtures for peroral peptide drug delivery. AB - Newly synthesised starch-g-poly(acrylic acid) copolymers and starch/poly(acrylic acid) mixtures were evaluated for their in vitro inhibition potency towards the proteolytic enzyme trypsin. Their Ca2+ and Zn2+ binding capacity was measured. Carbopol 934P was used as reference polymer. Starch-g-poly(acrylic acid) copolymers were prepared by chemical grafting and 60Co irradiation, the starch/poly(acrylic acid) mixtures by freeze-drying. The influence of preparation method, the ratio starch:acrylic acid, the neutralisation degree and for the freeze-dried polymers the influence of heat treatment after freeze-drying was investigated. All freeze-dried polymers showed a higher inhibition factor (IF) than the chemically grafted and 60Co irradiated starches, which all showed significantly lower IF than Carbopol 934P. The heat treated freeze-dried polymer Amioca/poly(acrylic acid) (1:1) showed a significantly higher IF than the reference polymer (Mann-Whitney test, p<0.05). The Ca2+ and Zn2+ binding capacity of all chemically grafted starches was much lower than for Carbopol 934P. Only the 60Co irradiated starches and freeze-dried polymers with ratio 1:3 approached the binding capacity of the reference polymer. The freeze-dried polymers showed the highest proteolytic enzyme inhibition potency. Freeze-drying and 60Co irradiation could result in the highest ion binding capacity. This combination of proteolytic enzyme inhibition activity and ion binding capacity makes these polymers hopeful excipients for successful oral peptide delivery. PMID- 11489323 TI - Biorecognizable HPMA copolymer-drug conjugates for colon-specific delivery of 9 aminocamptothecin. AB - N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer conjugates for colon-specific delivery of 9-aminocamptothecin (9-AC) were designed. They hold 9-AC bound via spacers containing amino acid residues and aromatic azo bonds. In vitro release profiles of 9-AC from HPMA copolymer conjugates were evaluated under artificial conditions that simulated large intestinal azoreductase and peptidase activities. The studies indicated that the azo bond was reduced first, followed by the release of unmodified 9-AC from the 9-AC containing fragment by peptidases. Release profiles depended on the chemical structure of the peptide part of the spacer. Conjugates containing leucylalanine showed high colon-specific release of 9-AC when compared to alanine containing conjugates. It appears that the studied conjugates are suitable as colon-specific drug delivery systems. PMID- 11489324 TI - Efficiency of liposomes surface-modified with soybean-derived sterylglucoside as a liver targeting carrier in HepG2 cells. AB - We investigated the interaction of liposomes surface-modified with soybean derived sterylglucoside (SG) (SG-liposomes) with HepG2 cells in the point of involvement of asialoglycoprotein receptor (ASGP-R) mediated endocytosis and examined the efficiency of SG-liposomes as drug carriers using 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) as a maker of liposome, carboxylated polystyrene microspheres (Fluoresbrite) as a model drug not taken up in cells and doxorubicin (DXR). SG-liposomes were composed of dipalmitoylphosphatidylcholine (DPPC), cholesterol (Ch) and SG (DPPC/Ch/SG=6:3:1, molar ratio) and DiI, Fluoresbrite and DXR were entrapped in SG-liposomes, respectively. Each SG-liposome was incubated with HepG2 cells at 4 or 37 degrees C, and co-incubated with asialofetuin (AF) as a competitor of ASGP-R. The association of DiI, Fluoresbrite or DXR entrapped in SG-liposomes with HepG2 cells at 37 degrees C was significantly higher than that in liposomes containing no SG. That of DiI and Fluoresbrite was reduced significantly by the incubation with AF, but that of DXR was not affected. These findings suggest that Fluoresbrite behaves like the lipid component of SG-liposomes, but DXR in SG liposomes does not behave similar to the lipid component of SG-liposomes, thus, its drug behavior released from liposomes may be due to its physicochemical properties. SG-liposomes are potentially useful drug carriers to the liver, because the glucose residue may work as a kind of ligand for ASGP-R. PMID- 11489325 TI - In vitro and in vivo evaluation of starch-based hot stage extruded double matrix systems. AB - The objective of developing a double matrix system consisting of a hot stage extruded starch pipe surrounding a hot stage extruded and drug-containing starch core, was to obtain a monolithic matrix system applicable in the domain of sustained drug release. The behaviour of the systems was evaluated through dissolution testing and through a randomised crossover bioavailability study on nine male volunteers. All double matrix systems showed in vitro a nearly constant drug release profile after an initial slower release phase of 4 h. This initial slower release phase was avoided by loading the starch pipe with a small amount of drug. The in vitro dissolution profiles did not reflect the in vivo behaviour of the systems. Although the in vitro dissolution profiles of two selected double matrix systems were slower than the profile of a commercially available sustained release formulation, they performed less well in vivo. However, the t75%Cmax ratios of the evaluated double matrix systems versus a reference immediate release formulation indicated an acceptable sustained release behaviour, superior to that of hot stage extruded starch-based single matrices. Therefore the disclosed drug delivery systems could be applied in the domain of sustained drug delivery. Besides, the continuous production of the double matrix systems offers an advantage from a manufacturing point of view. PMID- 11489326 TI - DQAsome/DNA complexes release DNA upon contact with isolated mouse liver mitochondria. AB - DQAsomes are mitochondriotropic cationic vesicles, which have been developed by us for the supposed transport of DNA to mitochondria in living cells [Pharm. Res. 15 (1998) 334]. Our strategy for the delivery of DNA into the matrix of mitochondria is based upon the putative transport of a DNA-signal peptide conjugate to mitochondria, the liberation of this conjugate from DQAsomes at the mitochondrial membrane followed by DNA uptake via the mitochondrial protein import machinery. As a first and important step towards delivery of DNA into mitochondria of living cells, we studied the DNA release from DQAsomes upon contact with non-energized mitochondria in vitro. Mitochondria were isolated from mouse liver and characterized by electron microscopy and the determination of mitochondrial marker enzyme activity. DQAsomes were added to DNA in the presence of SYBR Green I resulting in the formation of DQAsome/DNA complex and the complete loss of fluorescence. Following the addition of isolated mitochondria to DQAsome/DNA complex, the fluorescence signal was recovered due to the dissociation of DNA from its cationic carrier. Thus, DQAsome/DNA complexes were shown to release DNA upon contact with the surface of mitochondria thereby meeting a key requirement for our strategy towards mitochondrial DNA delivery. PMID- 11489327 TI - Chitosan-based nanoparticles for topical genetic immunization. AB - Numerous studies have reported the prophylactic and therapeutic use of genetic vaccines for combating a variety of infectious diseases in animal models. Recent human clinical studies with the gene gun have validated the concept of direct targeting of dendritic cells (Langerhan's cells) in the viable epidermis of the skin. However, it is unclear whether the gene gun technology or other needle-free devices will become commercially viable. The objective of our studies was to investigate the topical application of chitosan-based nanoparticles containing plasmid DNA (pDNA) as a potential approach to genetic immunization. Two types of nanoparticles were investigated: (i) pDNA-condensed chitosan nanoparticles, and (ii) pDNA-coated on pre-formed cationic chitosan/carboxymethylcellulose (CMC) nanoparticles. These studies showed that both chitosan and a chitosan oligomer can complex CMC to form stable cationic nanoparticles for subsequent pDNA coating. Selected pDNA-coated nanoparticles (with pDNA up to 400 microg/ml) were stable to challenge with serum. Several different chitosan-based nanoparticles containing pDNA resulted in both detectable and quantifiable levels of luciferase expression in mouse skin 24 h after topical application, and significant antigen specific IgG titer to expressed beta-galactosidase at 28 days. PMID- 11489328 TI - Cell type specific and glucose responsive expression of interleukin-4 by using insulin promoter and water soluble lipopolymer. AB - For gene therapy, tissue targeting of gene delivery systems is required for the maximum efficiency. In this study, we constructed pRIP-IL4 in which the expression of interleukin-4 (IL-4) was driven by the rat insulin promoter. WSLP pRIP-IL4 complex was characterized by pancreas beta-cell specific and glucose responsive expression of IL-4. pRIP-IL4 was completely retarded at a 6:1 or higher N/P (nitrogen atom of WSLP/phosphate of plasmid) ratio in 1% agarose gel. In addition, WSLP protected plasmid DNA from DNase I for more than 1 h. In cytotoxicity assay, WSLP showed less cytotoxicity than PEI (25000 Da) to mouse insulinoma (MIN6) cells. ELISA showed that pRIP-IL4 expressed much higher levels of IL-4 in MIN6 cells than in NIH3T3 cells. The expression level of IL-4 by pRIP IL4 increased with increasing concentration of glucose. Also, IL-4 was expressed in a dose-dependent manner. This WSLP-pRIP-IL4 system will be useful in the development of a pancreas specific expression system for the prevention of diabetes without systemic side effects. PMID- 11489330 TI - Transport and absorption of drugs via the lymphatic system. PMID- 11489331 TI - The physiology of the lymphatic system. AB - This paper presents an overview of the anatomy, physiology, and biology of the lymphatic system specifically relevant to lymphatic drug delivery. We will briefly review the classic fluid and solute transport literature, and also examine the current research in lymphatic endothelial cell biology and tumor metastasis in the lymphatics because of the increasing potential for targeted delivery of immunomodulators, chemotherapeutics, and genetic material to specific lymph nodes (Refs. [1-7]). PMID- 11489332 TI - An examination of the factors affecting intestinal lymphatic transport of dietary lipids. AB - Lipophilic drugs are carried by chylomicrons secreted by the small intestine and transported in lymph. The intent of this review is to update the reader on the digestion, uptake, and transport of dietary lipids and how these processes impact the absorption of lipophilic drugs by the gut. The digestion of lipids in the gastric and intestinal lumen is discussed as well as the role of bile salts in the solubilization of lipid digestion products for uptake by the gut. Both passive and active uptake of lipid digestion products is reviewed. Also examined is how intestinal lipid transporters located at the brush border membrane may play a role in the uptake of lipids by the enterocytes. The intracellular trafficking and the resynthesis of complex lipids from lipid digestion products are explored. Finally, the formation and secretion of chylomicrons and their potential clinical disorders are described. PMID- 11489333 TI - Animal models for the study of intestinal lymphatic drug transport. AB - Drug transport via the intestinal lymphatic system has been shown to contribute to the absorption of a number of orally administered highly lipophilic drugs. In order to investigate this phenomenon and assist in the development of improved oral formulations, the use of appropriate animal models is required. This paper reviews the use of various animal models for this purpose, and describes in detail the conscious rat and dog models used in our laboratory. The advantages and disadvantages of both small and large animal models are explored, as well as the factors which have been found to influence the outcome of intestinal lymphatic drug transport studies with these models. PMID- 11489334 TI - Intestinal lymphatic drug transport: an update. AB - The trend towards identification of poorly water-soluble and highly lipophilic candidate drug molecules has led to an increase in interest in intestinal lymphatic drug transport. In this article we provide a brief background to the mechanism of access of drugs to the intestinal lymph and the role of lipid digestion and absorption in the stimulation of lymphatic transport. The ability of different lipid types to stimulate lymphatic drug transport, is addressed, concentrating specifically on the impact of the class, chain length and degree of unsaturation of co-administered lipids. Comment is also made as to the relevance of dosing different lipid volumes to the rat and the possible complications this may provide when trying to assess the likely extent of intestinal lymphatic transport. Recent studies are described in which the extent of lymphatic transport of a highly lipophilic antimalarial, halofantrine, was investigated after post-prandial administration to greyhound dogs. Finally the possible future directions for studies of intestinal lymphatic transport are discussed, including the use of cell culture models and genetically modified animals. PMID- 11489335 TI - Exploiting M cells for drug and vaccine delivery. AB - The specialised antigen sampling M cells represent an efficient portal for mucosal drug and vaccine delivery. Delivery may be achieved using synthetic particulate delivery vehicles including poly(DL-lactide-co-glycolide) microparticles and liposomes. M cell interaction of these delivery vehicles is highly variable, and is determined by the physical properties of both particles and M cells. Delivery may be enhanced by coating with reagents including appropriate lectins, microbial adhesins and immunoglobulins which selectively bind to M cell surfaces. Live attenuated microorganisms are also suitable as vaccines and mucosal vectors and many, including Salmonella typhimurium, innately target to M cells. After cell surface adhesion, delivery vehicles are rapidly transported across the M cell cytoplasm to underlying lymphoid cells and may subsequently disseminate via the lymphatics. Further definition of M cell development and function should permit exploitation of their high transcytotic capacity for safe and reliable mucosal delivery. PMID- 11489336 TI - Recent advances in the understanding of uptake of microparticulates across the gastrointestinal lymphatics. PMID- 11489337 TI - Liposomes to target the lymphatics by subcutaneous administration. AB - Liposomes have been proposed as carriers for the delivery of therapeutic and diagnostic agents to the lymphatic system. Subcutaneous (s.c.) injection is the route of administration most extensively studied for this purpose. Decisive factors influencing lymphatic absorption and lymph node uptake of s.c. administered liposomes are liposome size and the anatomical site of injection. Generally, other factors such as lipid composition, charge and the presence of a hydrophilic PEG-coating on the liposome surface do not substantially affect lymphatic absorption and lymph node uptake of s.c. administered liposomes. Studies on the intranodal fate of liposomes demonstrate that phagocytosis by macrophages is the most important mechanism for lymph node uptake of liposomes. The observation of relatively high uptake of liposomes in regional lymph nodes after s.c. administration has stimulated research on lymphatic targeting of liposomes for diagnostic and therapeutic applications. PMID- 11489338 TI - Lymphatic transport of proteins after s.c. injection: implications of animal model selection. AB - Subcutaneous (s.c.) administration continues to be the main route for the delivery of protein drugs due to their poor bioavailability by most non parenteral routes. While small drug molecules are rapidly and extensively absorbed after s.c. injection, the systemic bioavailability of protein drugs is often incomplete and variable. Given the widespread use of the s.c. route for protein drugs, surprisingly little is known about the factors that govern the rate and extent of protein absorption from the interstitial space and the role of the lymphatic system in the transport of these molecules to the systemic circulation. The few studies that have directly addressed the role of lymphatic transport in protein bioavailability are complicated by the use of methods and models that vary widely. In this review we will evaluate the available literature describing the lymphatic transport of proteins after s.c. injection and more specifically, address the impact of experimental variation (e.g. site of cannulation, animal model, anesthesia) on the interpretation of the data obtained. We will also describe in some detail the sheep model currently in use in our laboratory, which allows both estimation of the extent of uptake of protein drugs into the lymphatics draining the injection site, and quantification of the contribution of lymphatic transport to the absolute bioavailability. PMID- 11489339 TI - Behavioral and biochemical effects of glutathione depletion in the rat brain. AB - Glutathione serves the function of providing reducing equivalents for the maintenance of oxidant homeostasis, and besides it plays roles in intra- and intercellular signaling in the brain. Our purpose was to test the effects of depleting tissue glutathione by diethylmaleate (5.3 mmol/kg, intraperitoneal) on brain antioxidant metabolism, nerve growth factor levels, and cognitive performance in rats. Six hours after the treatment, glutathione level in the hippocampus dropped down to 30% of the mean value of vehicle-treated animals and glutathione peroxidase activity also declined. Twenty-four hours after the injection the values had been partially restored. Moreover, the hippocampal and cortical levels of nerve growth factor protein did not change in response to diethylmaleate treatment. Glutathione depletion did not influence the performance of animals in the step-through passive avoidance test, but impairs acquisition in the Morris water maze when given before training. However, when diethylmaleate was administered after acquisition in the same paradigm, it did not affect the retention tested at the following day. Our results suggest that glutathione status is important during acquisition, but not for retention, of spatial memory in maze tasks and they support the hypothesis of the oxidant/antioxidant equilibrium as a key piece acting in the regulation of brain function. PMID- 11489340 TI - Nitric oxide inhibits GABA-evoked current in dorsal root ganglion neuron via PKG dependent pathway. AB - gamma-Aminobutyric acid (GABA) is considered a major inhibitory neurotransmitter in the generation of presynaptic inhibition at central terminals of primary afferent fiber (PAF), while it has also been established that nitric oxide (NO) may sensitize the terminals of nocisponsive PAFs and enhance neuropeptide release, possibly via mechanisms involving the activation of a cyclic guanidine monophosphate (cGMP)-dependent PKG. The present work was undertaken to explore the modulatory effect of sodium nitroprusside (SNP), a donor of NO, on GABA evoked current of isolated adult rat dorsal root ganglion (DRG) neurons and the intracellular mechanism involved, by means of whole-cell patch clamp recording. The results showed that 1 mM SNP reversibly inhibited the inward current evoked by 0.1 mM GABA (-1.05 +/- 0.17nA vs. -0.63 +/- 0.11nA, n = 22, p < 0.01 or 0.1 mM muscimol a specific GABA(A) receptor agonist (-1.70 +/- 0.39 nA vs. -1.01 +/- 0.24 nA, n = 6, p < 0.05), which could be cancelled by simultaneous application of 1 mM methylene blue, an inhibitor of PKG. After preapplication of SNP with increasing concentrations 0.03, 0.1, 0.3, 1, and 3 mM), SNP inhibited both 0.1 mM GABA-evoked current (IC(50) = 0.2423 mM, n = 5) and 0.1 mM muscimol-evoked current (IC(50) = 0.3255 mM, n = 3) in DRG neurons in a dose-dependent manner. Therefore, it was suggested that PKG-dependent pathway may be involved in the NO induced inhibition of GABA(A) receptor-mediated inward current in rat DRG neurons, which may be involved in the presynaptic disinhibition of nociceptive information induced by NO under certain conditions. PMID- 11489341 TI - Transplacental transfer of the opioid growth factor, [Met(5)]-enkephalin, in rats. AB - Placental transfer of the pentapeptide [Met5]-enkephalin, known to function as a growth regulating factor and neuromodulatory agent, was studied in pregnant Sprague-Dawley rats. Using separation by reversed phase high-performance liquid chromatography, and analysis by derivative spectroscopy, [Met5]-enkephalin was detected in 20-day-old fetal tissue including brain, heart, lung, and kidney. Fetal tissues from pregnant rats given an injection of 40 mg/kg [Met5]-enkephalin on gestation day 20 had markedly elevated levels of peptide within 1 h, indicating the transplacental transfer of this opioid. [Met5]-enkephalin levels were increased from control samples at 1, 2, 4, and 14 h post-injection of peptide, but not at 24 h. Evaluation of breakdown products of [Met5]-enkephalin, along with the related peptide [Leu5]-enkephalin, revealed that elution times differed substantially from [Met5]-enkephalin. These data indicate that [Met5] enkephalin is present in fetal organs, crosses the placenta, does not appear to be restrictive in organ specificity, and is sustained in fetal tissues at detectable levels for at least 14 h. Given that [Met5]-enkephalin tonically inhibits DNA synthesis in the fetus, these results raise the question of whether an elevated level of this peptide (either maternally or from the fetus) may be detrimental to cellular ontogeny in the fetus, and perhaps have long-term implications for postnatal development. PMID- 11489342 TI - Alterations of AP-1 and CREB protein DNA binding in rat supraoptic and paraventricular nuclei by acute and repeated hyperosmotic stress. AB - Electrophoretic mobility shift assays were used to analyze Fos and CREB protein DNA-interactions in the rat hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. After intraperitoneal administration of normal saline, PVN (but not SON) extracts exhibited a significant 183% increase in binding to the activational protein-1 (AP-1) canonical DNA binding sequence. Hypertonic saline treatment resulted in a approximately 2.5-fold increase in binding by tissue samples from both regions. AP-1 binding by SON extracts after two hypertonic saline injections caused a 307% increase in binding that was significantly greater than binding by PVN extracts (207%). Fos binding was equal in the SON after one and two hypertonic saline injections, but the PVN exhibited less of an increase after two injections. Binding to the canonical cyclic adenosine monophosphate regulatory element (CRE), and phosphorylated CREB (pCREB) supershift binding, indicated pCREB is constitutively expressed. Any experimental treatment (handling and an injection) caused an elevation in binding in the PVN. AP-1 protein complex DNA binding was increased after osmotic stimulation, and SON and PVN exhibit differences in AP-1 DNA binding kinetics, after repeated hypertonic saline stress. Changes in PVN tissue samples were subtle, and may reflect the fact that magnocellular and parvocellular neurons mediate, respectively, fluid homeostasis and stress responses. PMID- 11489343 TI - Lateral parabrachial nucleus lesions in the rat: neophobia and conditioned taste aversion. AB - The present study investigated the hypothesis that the conditioned taste aversion (CTA) deficit consequent to lesions of the lateral parabrachial nucleus (LPBN) may be due to a disruption of neophobia. In Experiment 1, subjects were tested with one of three taste stimuli (alanine, saccharin, or quinine) and two nontaste stimuli (capsaicin and almond odor). Ibotenic acid lesions of the LPBN eliminated neophobia to alanine and saccharin but had no influence on the neophobic response to quinine, capsaicin, or almond odor. In Experiment 2, all the LPBN-lesioned (LPBNX) rats failed to develop a CTA. These results do not support the experimental hypothesis. Not only was the lesion-induced disruption of neophobia restricted to taste stimuli, the deficit was selective within that category. It is already known that LPBNX rats are unable to acquire conditioned aversions to capsaicin as well as alanine. Thus, the absence of a conditioned ingestional aversion in LPBNX rats is not predicated upon the absence of a neophobic response to the target stimulus. The present results, although exposing a stimulus selective disruption of neophobia, suggest that this deficit is independent of, rather than responsible for, the absence of conditioned ingestional aversions in rats with LPBN lesions. PMID- 11489344 TI - Regional distribution of Hsp70 in the CNS of young and old food-restricted rats following hyperthermia. AB - We examined the effect of aging on the capacity of the brain to produce heat shock protein (Hsp70) in response to heat stress, using high-powered microwaves (HPM, 2.06 GHz, 2.2 W/cm(2)) to induce hyperthermia for periods so brief that thermoregulatory factors were functionally eliminated as confounding variables. Unanesthetized young (6 months) and old (25 months) male, food-restricted Sprague Dawley rats were exposed to HPM to induce a mean peak tympanic temperature (T(ty)) of 42.2 degrees C within 30 s. T(ty) returned to <40.0 degrees C within 6 min post-exposure in both age groups. Rats were euthanized 6 or 24 h later for immunohistochemical determination of Hsp70 accumulation in 10 brain regions. HPM exposure induced significant increases in 7 of the 10 regions. There were no significant differences observed in the pattern or density of Hsp70 accumulation between the young and old rats at 6 h post-HPM exposure, with the exception of the medial vestibular nucleus, which demonstrated significantly greater Hsp70 accumulation in the old rats. There were significant differences between the age groups at 24 h post-exposure, however, there was no general pattern; i.e., depending on the brain region, aged rats displayed significantly greater, lesser, or similar increases in Hsp70 expression compared with young. Taken together, these results demonstrate that the brain of aged, food-restricted rats does not display a loss of capacity to accumulate Hsp70 in response to heat stress. PMID- 11489345 TI - The NF-kappaB inhibitor diethyldithiocarbamate (DDTC) increases brain cell death in a transient middle cerebral artery occlusion model of ischemia. AB - A transient ischemic middle cerebral artery occlusion model of stroke was used to examine the role of the transcription factor NF-kappaB in cell death as measured by DNA fragmentation and infarction volume. The left middle cerebral artery was occluded for either 30 min or 2 h in rats. One set of animals was pretreated with diethyldithiocarbamate (DDTC), an inhibitor of NF-kappaB, 30 min prior to reperfusion. The animals were reperfused and allowed to survive for 2 or 7 days. DNA fragmentation was assayed by in situ end labeling in the stroke core and penumbral regions. Specific cortical and subcortical regions were measured using quantitative image analysis. DNA fragmentation was seen only on the ischemic side of the brains in all cases. Overall, the DDTC-treated groups showed significantly increased DNA fragmentation within the ischemic side compared to the saline control groups. DDTC treatment also caused an increase in stroke volume based on triphenyl tetrazolium chloride staining. Electrophoretic mobility shift assays showed NF-kappaB activation peaking 15 min following reperfusion and that this activation was blocked by the DDTC treatment. This study suggests that the use of NF-kappaB inhibitors to block cell death following stroke needs to be carefully examined because global inhibitors may not promote neuronal survival. PMID- 11489346 TI - Comparison of anticonvulsive and acute neuroprotective activity of three 2,3 benzodiazepine compounds, GYKI 52466, GYKI 53405, and GYKI 53655. AB - GYKI 52466 [1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine], a non-competitive AMPA [alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate] and kainate receptor antagonist and its two analogues, GYKI 53405 [1-(4-aminophenyl) 3-acetyl-4-methyl-3,4-dihydro-7,8-methylenedioxy-5H-2,3-benzodiazepine] and GYKI 53655 [1-(4-aminophenyl)-3-methylcarbamyl-4-methyl-3,4-dihydro-7,8-methylenedioxy 5H-2,3-benzodiazepine] were investigated in two seizure models and in MgCl2 induced global cerebral ischaemia, as an acute neuroprotective model. The ED(50) values of GYKI 52466 for suppression of the tonic and clonic phases of sound induced seizures were 3.6 and 4.3 mg/kg, respectively. The corresponding data for GYKI 53405 were 1.1 and 3.1 mg/kg, while ED(50) values of GYKI 53655 were 1.3 and 2.0 mg/kg, respectively. The inhibition of seizure evoked by maximal electroshock was also found to be remarkable: the ED(50) values of GYKI 52466 and its two analogues were 6.9, 2.6, and 2.2 mg/kg, respectively. All compounds prolonged the survival times in MgCl2 induced global cerebral ischaemia test in a dose dependent fashion, with PD(50) (dose of 50% prolongation) values of 24.1, 8.3, and 8.2 mg/kg intraperitoneal, respectively. In audiogenic seizure model the duration of anticonvulsant action of 10 mg/kg GYKI 52466 and 5 mg/kg GYKI 53405, GYKI 53655 were examined, too. The effect of GYKI 52466 decreased to 50% after 2 h, while the analogues showed more than 80% seizure suppression 3 h after treatment. After 6 h the effect of GYKI 53655 decreased to zero, while the effect of GYKI 52466, remained on the 50% level. PMID- 11489347 TI - An experimental study on the course of trans-synaptic propagation of neural activity and plasticity in the hippocampus in kainate-induced epilepsy. AB - To investigate the course of trans-synaptic propagation of neural activity and plasticity in temporal lobe epilepsy, time-dependent changes in the level of synapsin I, a synaptic vesicle protein that is a marker of enhanced synaptic activity and synaptogenesis, were examined following kainate-induced epileptic status in rats. Compared with the control, the level of synapsin I protein increased in the bilateral stratum oriens of CA3 (28.8-40.2%) and CA1 (28.0 34.6%), and the stratum radiatum of CA1 (34.0%) ipsilateral to the injection site at 8 h after intra-amygdala administration of kainate. At 24 h, and 2 and 4 weeks after the kainate treatment, however, synapsin I levels returned to normal levels in most of the regions studied in spite of the extended neural loss in the hippocampus, which suggested the axonal sprouting on the remaining cells. The synapsin I mRNA levels time-dependently decreased bilaterally in CA1-CA3 and the hilus, while no significant changes were observed in the dentate gyrus. These results suggest that the synaptic input to CA3 and CA1 through the stratum oriens was enhanced in this model. A different mode of hippocampal neural activity and plasticity between kainate and kindling models of epilepsy may be stressed. PMID- 11489348 TI - Differential distribution of Fos expression within the male rat preoptic area and hypothalamus in response to physical vs. psychological stress. AB - Recent studies on stress-induced adrenal glucocorticoid secretion have demonstrated quantitatively different effects of individual stress stimuli on hormone release, suggesting that the hypothalamic-pituitary-adrenal axis exhibits discriminative, rather than ubiquitous responses to such challenges, particularly psychological vs. physical stressors. The immediate-early gene, cfos, is expressed by central nervous system neurons in response to numerous physiological stimuli, including stress. The following study investigated whether the distribution and/or intensity of immunolabeling for Fos in the preoptic area and hypothalamus differ after imposition of stressors of variable intensity. Groups of male rats were sacrificed by transcardial perfusion 2 h after (1) transfer to a novel environment (NE stress), (2) confinement within a restraint tube (REST stress), or (3) immobilization (IM) stress. Nonstressed controls remained undisturbed in their home cages. Whereas the NE-stress group exhibited Fos immunoreactivity (ir) only within anterior and lateroanterior hypothalamus, both physical stressors induced immunostaining for Fos in the lateral preoptic area, median preoptic, paraventricular, arcuate, dorsomedial hypothalamic nuclei, and lateral hypothalamus, while numbers of Fos-ir-positive neurons were generally greater in the IM vs. REST stress group. In the IM-stressed rats, additional Fos ir was observed in the supraoptic and suprachiasmatic nuclei. These studies show that neurons expressing Fos-ir in response to the relatively mild stress of novel environment are distributed differently than those that are transcriptionally activated by more aversive stressors. Findings that Fos-ir occurs in several common sites following exposure to REST or IM stress suggest that local neuron populations may comprise a common neural pathway(s) that is activated by intense forms of stress. PMID- 11489349 TI - Optic nerve regeneration within artificial Schwann cell graft in the adult rat. AB - We investigate whether an artificial graft made by cultured Schwann cell, extracellular matrix (ECM) and trophic factors can provide the environment for the regeneration of retinal ganglion cell (RGC) axons in adult rats. Six kinds of artificial grafts were used: ECM (control); ECM and Schwann cells; ECM, Schwann cells and either nerve growth factor, brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4); ECM, Schwann cells, BDNF and NT-4, combined with intravitreal injection of BDNF. The grafts were transplanted onto the transected optic nerve. RGC regeneration was evaluated by dil retrograde labeling, immunohistochemistry, and electron microscopy at 3 weeks post-operation. The degree of dil labeled RGC was approximately 2% for ECM alone, and 10% for ECM and Schwann cells (p < 0.01). The labeling increased to approximately 20% by administration of neurotrophins. The addition of intravitreous BDNF injection resulted in highest labeling percentage of 30%. Immunohistochemical study showed that axons were association with GAP-43 and cell adhesion molecules. Neurotrophin receptors (Trk-A and Trk-B) were detected in nerve fibers both in the retina and in the graft. Remyelination was seen by electron microscopic observation. These results demonstrate that the regeneration of RGC axons is induced with the use of cultured Schwann cells and ECM as promoting factors for regrowth. The degree of regeneration was significantly increased by neurotrophins in the grafts and in the vitreous. PMID- 11489350 TI - Changes in calcium currents and GABAergic spontaneous activity in cultured rat hippocampal neurons after a neurotropic influenza A virus infection. AB - In order to study mechanisms by which a neurotropic strain of influenza A virus (A/WSN/33) may affect neuronal function or cause nerve cell death, hippocampal cultures from embryonic rats were infected with this virus. Approximately 70% of the neurons in the infected cultures became immunopositive for viral antigens and showed reduced voltage-dependent Ca(2+) currents in whole-cell patch clamp recordings, but no changes in other membrane properties or in cytosolic Ca(2+) concentration were seen. These immunopositive neurons underwent apoptosis 3-4 days after infection. Ca(2+) channel inhibitors had no significant effect on neuronal survival. The immunonegative population of neurons survived, but displayed increased frequency of miniature inhibitory postsynaptic currents of gamma-amino-butyric acid origin compared with controls. The frequency of alpha amino-hydroxy-5-methylisoxazole-4-propionic acid hydrobromide (AMPA) receptor mediated miniature excitatory postsynaptic currents was not altered. Viral nucleoproteins, overexpressed using the Semliki Forest virus system, were localized to the dendritic spines as shown by double immunolabeling with actinin, but did not by themselves cause neuronal death or changes in synaptic transmission as measured by AMPA-mediated excitatory postsynaptic currents. Our results show that an influenza A virus infection can cause selective neurophysiological changes in hippocampal neurons and that these can persist even after the viral antigens have been cleared. PMID- 11489351 TI - Serotonergic innervation of the primate claustrum. AB - The claustrum is reciprocally and topographically connected with all functional areas of the cerebral cortex. Different cortical areas differ in the source, density, and laminar distribution of serotonergic innervation, with visual cortex receiving an especially rich serotonergic innervation. We asked if there were likewise differences in serotonergic innervation in different regions of the claustrum. We analyzed 50-microm coronal sections through the claustrum of the macaque monkey processed using standard immunohistochemical techniques and an antibody to serotonin. We found labeled fibers throughout the dorsal-ventral and anterior-posterior extent of the claustrum. A few fibers were relatively straight and lacked varicosities. Most fibers had varicosities; the size, shape, and spacing of varicosities varied among fibers and even along a single fiber. Some stained fibers partially encircled cells, and varicosities were seen in close apposition to the cell bodies. There was a major difference between dorsal and ventral claustrum in the pattern of stained fibers. In the ventral, visual, claustrum, stained segments of axons were short and randomly arranged relative to each other, and there were many stained puncta. In the more dorsal, nonvisual claustrum, many fibers ran in a dorsal-ventral direction, along the long axis of the claustrum, and could be followed for long distances. PMID- 11489352 TI - Age-associated changes in the serotonergic system in rat superior colliculus and pretectum. AB - The purpose of this study was to investigate whether aging alters serotonergic innervation of the superior colliculus and pretectum in rats. The superior colliculus has one of the highest concentrations of serotonin in the rat central nervous system. Young and old male F344 rats (<6 months, and >18 months, albino and pigmented) were used in all experiments. Coronal sections through the superior colliculus and pretectum were incubated with antibodies to serotonin, the serotonin 2A receptor, and the serotonin transporter. Immunocytochemical staining was analyzed semi-quantitatively. The results indicate that with age there is an increase in serotonin immunoreactivity throughout the entire superior colliculus and pretectum, a decrease in levels of serotonin 2A receptor staining in select layers of superior colliculus, and no change in serotonin transporter immunoreactivity. Albino rats differ from pigmented rats in that they have enhanced serotonergic immunoreactivity in the superficial layers of superior colliculus, a region that receives direct retinal input. These data suggest that the age-related changes in the serotonergic system in the superior colliculus and pretectum may account for some of the alterations in light-mediated behaviors with aging. PMID- 11489353 TI - Imiquimod-elicited emesis is mediated by the area postrema, but not by direct neuronal activation. AB - The immunomodulator, imiquimod, has been shown to have antiviral and antitumor properties in animal models. It also has been reported to alter cytokine levels in both animals and humans. However, because imiquimod appeared to be emetic, studies were conducted to determine the degree of sensitivity, and the pathways involved. Subcutaneous administration of > or = 10 mg/kg imiquimod to ferrets elicited emesis with latencies as short as 2'; 12 and 15 mg/kg were optimal doses. Emetic responsiveness was eliminated by complete ablation of the area postrema, but was unaffected by bilateral supradiaphragmatic section of the vagus nerve. This indicates that the emesis is produced by an activation of the chemoreceptor trigger zone B the area postrema. Ferret brain stem slices (450 microm) were preincubated in oxygenated Krebs-Ringer and then mounted in a submerged slice recording chamber. Extracellular recordings of spontaneous and ionophoretically evoked activity of area postrema neurons were obtained for up to 8 h, while the effect of bath-applied imiquimod was determined. Under control conditions, neurons showed a low frequency spontaneous discharge. Introduction of imiquimod (concentration range, 1 x 10(-7) to 5 x 10(-8)M) had no effect on neuronal firing. With ionophoresis of glutamate from an independent micropipette, a brief excitatory response was obtained. We conclude that imiquimod does not directly excite area postrema neurons. It is likely that imiquimod causes synthesis and release of some unknown emetic substance(s), very possibly cytokines. PMID- 11489354 TI - Use of cDNA microarrays to analyze dioxin-induced changes in human liver gene expression. AB - One mechanism by which cells adapt to environmental changes is by altering gene expression. Here, we have used cDNA microarrays to identify genes whose expression is altered by exposure to the environmental contaminant 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). The goal of our study was to enhance our understanding of toxicity mediated through the pathway by which TCDD stimulates gene expression. To model this toxicity response, we exposed human hepatoma (HepG2) cells to TCDD (10 nM for 18 h) and analyzed mRNA by two-color fluorescent hybridization to cDNA sequences immobilized on glass microscope slides (2.5 x 7.5 cm) covering a surface area of 2.25 cm(2). We analyzed approximately one-third of the genes expressed in HepG2 cells and found that TCDD up- or down-regulates 112 genes two-fold or more. Most changes are relatively subtle (two- to four-fold). We verified the regulation of protooncogene cot, XMP, and human enhancer of filamentation-1 (HEF1), genes involved in cellular proliferation, as well as metallothionein, plasminogen activator inhibitor (PAI1), and HM74, genes involved in cellular signaling and regeneration. To characterize the response in more detail, we performed time-course, dose-dependence studies, and cycloheximide experiments. We observed direct and indirect responses to TCDD implying that adaptation to TCDD (and other related environmental stimuli) is substantially more complex than we previously realized. PMID- 11489355 TI - H(2)O(2) mediates oxidative stress-induced epidermal growth factor receptor phosphorylation. AB - We used a well-established thiol-alkylating agent, N-ethylmaleimide (NEM), to oxidatively stress human keratinocytes. Time course studies revealed that NEM rapidly depleted keratinocytes of reduced glutathione (GSH), which was followed by rapidly increasing levels of intracellular reactive oxygen species (ROS) and subsequently by phosphorylation of epidermal growth factor receptor (EGFR). Pretreatment with antioxidants or enhanced catalase activity in keratinocytes inhibited ROS/H(2)O(2) accumulation and EGFR phosphorylation, demonstrating that H(2)O(2) production is a mediator required for EGFR phosphorylation. Collectively, these results suggest a sequence of events leading to EGFR phosphorylation which is likely shared by oxidative stress-inducing agents, namely: (1) GSH depletion; (2) H(2)O(2) accumulation; and (3) EGFR phosphorylation. We propose that depletion of GSH and accumulation of H(2)O(2) are upstream events and critical mediators required for ligand-independent phosphorylation of growth factor receptors in response to oxidative stress. PMID- 11489356 TI - GABA receptor subunit composition relative to insecticide potency and selectivity. AB - Three observations on the 4-[(3)H]propyl-4'-ethynylbicycloorthobenzoate ([(3)H]EBOB) binding site in the gamma-aminobutyric acid (GABA) receptor indicate the specific target for insecticide action in human brain and a possible mechanism for selectivity. First, from published data, alpha-endosulfan, lindane and fipronil compete for the [(3)H]EBOB binding site with affinities of 0.3--7 nM in both human recombinant homooligomeric beta 3 receptors and housefly head membranes. Second, from structure-activity studies, including new data, GABAergic insecticide binding potency on the pentameric receptor formed from the beta 3 subunit correlates well with that on the housefly receptor (r=0.88, n=20). This conserved inhibitor specificity is consistent with known sequence homologies in the housefly GABA receptor and the human GABA(A) receptor beta 3 subunit. Third, as mostly new findings, various combinations of alpha 1, alpha 6, and gamma 2 subunits coexpressed with a beta 1 or beta 3 subunit confer differential insecticide binding sensitivity, particularly to fipronil, indicating that subunit composition is a major factor in insecticide selectivity. PMID- 11489357 TI - Both hypomethylation and hypermethylation of DNA associated with arsenite exposure in cultures of human cells identified by methylation-sensitive arbitrarily-primed PCR. AB - In a previous study we reported that methylation within the promoter region of p53 was altered in human lung A549 cells exposed to arsenite over a 2-week period in culture. In the present study the methylation status of the 5' control region of the tumor suppressor gene, von Hippel Lindau syndrome (VHL), a gene known to be silenced transcriptionally by CpG methylation was assessed. No changes in DNA methylation in VHL in human kidney UOK cell lines exposed to arsenite were seen after 4 weeks in culture, assessed by simple HpaII digestion followed by PCR amplification. Using methylation-sensitive arbitrarily-primed PCR we identified eight differentially methylated regions of genomic DNA of approximately 300--500 bp from three UOK cell lines and from human lung A549 cells after arsenite exposure in culture. Six fragments were hypermethylated, and two were hypomethylated, relative to untreated controls. Sequence analysis revealed two DNA fragments contained repeat sequences of mammalian-apparent LTR retrotransposons, five contained promoter-like sequences, and 13 CpG islands were identified. Three fragments had 99-100% homology to regions on human chromosomes 6, 9, and 15 but these genes have not yet been identified. Our findings are consistent with a potential role for both hypermethylation and hypomethylation of DNA that coexist after exposure to arsenite. The results, in total, could support the existence of a state of DNA methylation imbalance that could conceivably disrupt appropriate gene expression in arsenite exposed cells. PMID- 11489358 TI - Effects of different nickel compounds on the transport of para-aminohippurate ion by rat renal cortical slices. AB - In vitro rat renal cortical slice uptake of para-aminohippurate ion (PAH) expressed as tissue slice-to-medium ratio was used as a model to predict nephrotoxicity of different nickel compounds. Pretreatment with nickel compounds for a period of up to 4 h was followed by an incubation with 74 microM PAH for 2 h. The PAH uptake by renal cortical slices was reduced by nickel compounds in both concentration- and time-dependent manner. Furthermore, the extent of such reduction of PAH uptake was found to depend on the chemical form (speciation) of nickel. Thus, slice-to-medium ratios were 41, 49, 48, and 66% of control values after a 4-h pretreatment with 0.378 mM nickel carbonate hydroxide, 1.5 mM nickel subsulfide, 4 mM nickel sulfate and 2.98 mM nickel oxide, respectively. Such an inhibition of PAH transport was not due to cytotoxicity. The results suggest that the nephrotoxic potency decreases in the following order: nickel carbonate hydroxide>nickel subsulfide>nickel sulfate>nickel oxide. Treatment of renal cortical slices with high concentrations of either mannitol, a hydroxyl radical scavenger, or glutathione significantly reduced the inhibition of PAH uptake by soluble form of nickel subsulfide, with concomitant reduction of nickel ion uptake by cortical slices. But no such oxidative stress seems to be involved in nickel carbonate hydroxide-induced inhibition of PAH uptake. PMID- 11489359 TI - Mechanisms of diethylstilbestrol-induced calcium movement in MG63 human osteosarcoma cells. AB - The effect of the estrogen diethylstilbestrol (DES) on cytosolic free Ca(2+) levels ([Ca(2+)](i)) in MG63 human osteoblasts was explored by using fura-2 as a Ca(2+) indicator. DES at concentrations between 5--20 microM induced an immediate increase in [Ca(2+)](i) in a concentration-dependent manner with an EC(50) of 10 microM. Removing extracellular Ca(2+) reduced the Ca(2+) signal by 70%. Pretreatment with 50 microM La(3+) or 10 microM of nifedipine, verapamil and diltiazem did not change 20 microM DES-induced [Ca(2+)](i) increases. Addition of 3 mM Ca(2+) increased [Ca(2+)](i) in cells pretreated with 20 microM DES in Ca(2+)-free medium. Pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor) to deplete the endoplasmic reticulum Ca(2+) store partly inhibited 20 microM DES-induced Ca(2+) release, but addition of carbonylcyanide m-chlorophenylhydrazone (CCCP; a mitochondrial uncoupler) and thapsigargin together abolished DES-induced Ca(2+) release. Conversely, pretreatment with 20 microM DES abrogated CCCP- and thapsigargin-induced Ca(2+) release. Inhibition of phospholipase C activity with 2 microM U73122 did not alter 20 microM DES-induced Ca2+ release. Another estrogen 17beta-estradiol also increased [Ca(2+)](i) in a concentration-dependent manner with an EC50 of 7 microM. Together, the data indicate that in human osteoblasts, DES increased [Ca(2+)](i) via causing Ca(2+) release from both mitochondria and the endoplasmic reticulum in a phospholipase C-independent manner, and by causing Ca(2+) influx. PMID- 11489360 TI - LLU-alpha, an endogenous metabolite of gamma-tocopherol, is more effective against metal nephrotoxicity in rats than gamma-tocopherol. AB - Antioxidants of the vitamin E family have protective effects against metal toxicity. We examined the protective effect of racemic LLU-alpha [2,7,8-trimethyl 2-(carboxyethyl)-6-hydroxychroman] a metabolite of gamma-tocopherol, in comparison to the effect of alpha- and gamma-tocopherol in rats treated with sodium dichromate (Cr) or thallium sulfate (Tl). We measured metal nephrotoxicity based on urinary protein excretion and discussed it with respect to the metal concentration in renal tissue. The ranking of antioxidant activity (iron stimulated lipid peroxidation, luminol and lucigenin amplified chemiluminescence) was determined in the following order: alpha-tocopherol1000 microM). None of the compounds interacted with NMDA or AMPA receptors or with sodium or calcium (N, P/Q) channels. Brain penetration of both enantiomers following their i.p. administration was excellent, with brain and plasma concentrations being similar. Clomethiazole dose-dependently inhibited spontaneous locomotor activity in rats and was approximately 10 times more sedative than either enantiomer of AR-A008055. Clomethiazole was more potent than (S)-(-)-AR-A008055 in the "pull-up" test (muscle relaxation) and in producing loss of righting reflex, while (R)-(+)-AR-A008055 had little effect. The time animals remained on a Rota-rod was of the order: clomethiazole<(S)-(-)-AR A008055<(R)-(+)-AR-A008055. (S)-(-)-AR-A008055 (210 micromol/kg) raised seizure threshold to pentylenetetrazole (i.v.) by 119+/-21%. The (R)-(+)- enantiomer was not anticonvulsant. Overall, (S)-(-)-AR-A008055 exhibited a similar pharmacology to clomethiazole. However, its sedative and muscle relaxant effects were substantially less than clomethiazole, emphasising that these properties are not directly related to neuroprotective efficacy. The current data suggest that the proposed GABA uptake inhibitory property of (R)-(+)-AR-A008055 fails to produce significant sedative, myorelaxant or anticonvulsant activity. PMID- 11489454 TI - 5-HT1A receptor-mediated activation of G-protein-gated inwardly rectifying K+ current in rat periaqueductal gray neurons. AB - 5-hydroxytryptamine (5-HT) has been reported to modulate analgesia produced by opioids or electrical stimulation of the periaqueductal gray (PAG). 5-HT increases K+ conductance and inhibits the firing activity of the PAG neurons. We examined the electrophysiological and pharmacological characteristics of the K+ current involved in 5-HT-induced hyperpolarization of dissociated rat PAG neurons. Among the neurons tested, 5-HT activated inward K+ currents in 30-40%, whilst the remaining 60-70% did not respond to 5-HT. 5-HT activated an inwardly rectifying K+ current (I5-HT) in a concentration- and voltage-dependent manner. I5-HT was mimicked by a 5-HT1A receptor selective agonist, 8-OH-DPAT, and was reversibly blocked by a 5-HT1A receptor antagonist, piperazine maleate, but not by a 5-HT2 receptor antagonist, ketanserin. I5-HT was sensitive to K+ channel blockers such as quinine and Ba2+, but insensitive to 4-aminopyridine, Cs+ and tetraethylammonium. I5-HT was inhibited by GDP(beta)s and was irreversibly activated by GTP(gamma)s. I5-HT was significantly suppressed by N-ethylmaleimide and pertussis toxin, but not by cholera toxin. Second messenger modulators such as staurosporin, forskolin, and phorbol-12-myristate-13-acetate did not alter I5 HT. The present study indicates that 5-HT-induced hyperpolarization of the PAG neurons results from activation of the pertussis toxin-sensitive G-protein coupled inwardly rectifying K+ currents through 5-HT1A receptors. PMID- 11489455 TI - SB-243213; a selective 5-HT2C receptor inverse agonist with improved anxiolytic profile: lack of tolerance and withdrawal anxiety. AB - SB-243213 (5-methyl-1-[[-2-[(2-methyl-3-pyridyl)oxy]-5-pyridyl]carbamoyl]-6 trifluoromethylindoline hydrochloride) is a new, selective 5-hydroxytryptamine (5 HT)2C receptor inverse agonist. SB-243213 has high affinity for the human 5-HT2C receptor (pK(i) 9.37) and greater than a 100-fold selectivity over a wide range of neurotransmitter receptors, enzymes and ion channels. In in vitro functional studies, SB-243213 acted as an inverse agonist at the human 5-HT2C receptor with a pK(b) of 9.8. In in vivo studies, SB-243213 was a potent inhibitor of central 5 HT2C receptor-mediated function in rats, blocking meta-chlorophenylpiperazine induced hypolocomotion with an ID50 of 1.1 mg/kg p.o. and a long duration of action (>8 h). In rats, SB-243213 exhibited anxiolytic-like activity in both the social interaction and Geller-Seifter conflict tests. Importantly, unlike diazepam, chronic administration of SB-243213 did not result in the development of either tolerance to the anxiolytic-like effects or withdrawal anxiogenesis. Furthermore, in rodents, SB-243213 did not affect seizure threshold, did not increase body weight or induce catalepsy, but attenuated the haloperidol-induced catalepsy. SB-243213 did not affect amphetamine-, MK-801- or phencyclidine induced hyperactivity. In conclusion, SB-243213 may possess an improved anxiolytic profile compared to benzodiazepines. SB-243213 also modulates dopaminergic transmission, lacks pro-psychotic properties and may have utility in the treatment of schizophrenia and motor disorders. PMID- 11489456 TI - Evidence that hypophagia induced by d-fenfluramine and d-norfenfluramine in the rat is mediated by 5-HT2C receptors. AB - The present series of studies is the first to investigate the pharmacological mechanisms underlying d-fenfluramine- and d-norfenfluramine-induced hypophagia in the rat using highly selective serotonin 5-HT2 receptor antagonists. Administration of d-fenfluramine, and its major metabolite d-norfenfluramine, suppresses food intake in animals. Both compounds stimulate the release of serotonin and are potent inhibitors of the re-uptake of 5-HT into nerve terminals. In addition, d-norfenfluramine also acts as a direct 5-HT(2B/2C) receptor agonist. Pre-treatment with the selective 5-HT2C receptor antagonist, SB 242084 (0.3-3 mg/kg), dose-dependently inhibited both d-fenfluramine- (3 mg/kg) and d-norfenfluramine-induced (2 mg/kg) hypophagia. In contrast, the hypophagic effect of d-fenfluramine and d-norfenfluramine was unaffected by prior treatment with the highly selective 5-HT2B receptor antagonists, SB-215505 (0.3-3 mg/kg) and RS-127445 (1-3 mg/kg) or the 5-HT2A receptor antagonists MDL 100,907 (0.003 0.03 mg/kg) and ketanserin (0.2, 0.5 mg/kg). In addition, the 5-HT1A receptor antagonist WAY-100635 (0.3, 1 mg/kg) and the 5-HT1B receptor antagonists GR 127935 (1, 2 mg/kg) and SB-224289 (2-10 mg/kg) did not affect d-fenfluramine induced hypophagia. These data provide unequivocal evidence for an important role of the 5-HT2C receptor in the mediation of d-fenfluramine and d-norfenfluramine induced hypophagia in the rat and do not support the involvement of 5 HT1A/1B/2A/2B receptors. PMID- 11489457 TI - A role for 5-ht6 receptors in retention of spatial learning in the Morris water maze. AB - This study investigates the effect of intracerebroventricular administration of a 5-ht6 antisense oligonucleotide (AO) complementary to bases 1-18 of the rat 5-ht6 cDNA initiation sequence (Mol. Pharmacol. 43 (1993) 320) (1.5 microg twice daily for six days) and i.p. injection of a selective 5-ht6 receptor antagonist Ro 04 6790 (10 or 30 mg/kg once daily for three days) on acquisition and retention in the Morris water maze. Neither the 5-ht6 AO (which reduced cortical [3H]-LSD binding sites by 10-16%) nor Ro 04-6790 affected acquisition, but both enhanced retention of the learned platform position such that rats spent significantly longer searching the trained platform position than any other area during the probe tests. Furthermore, neither AO nor Ro 04-6790 had any effect on the time taken to reach a raised visible platform, indicating that visual acuity was unimpaired. In addition, AO reduced both food consumption and body weight and the later effect was also seen following Ro 04-6790, suggesting a role for the 5-ht6 receptor in the regulation of feeding. Hence, while the underlying mechanism remains unclear, enhanced retention of spatial learning following both AO and 5 ht6 antagonist administration strongly indicate a role for this receptor in memory processes. PMID- 11489458 TI - Slowing of ERG current deactivation in NG108-15 cells by the histidine-specific reagent diethylpyrocarbonate. AB - The aim of this study was to explore and characterize the effect of the histidine specific reagent diethylpyrocarbonate (DEPC) on the ERG (ether-a-go-go related gene) channels of whole-cell voltage-clamped NG108-15 neuroblastoma x glioma hybrid cells. The channels were fully activated by long depolarizing prepulses. Hyperpolarizing pulses elicited K+ inward currents which deactivated after reaching a peak. DEPC (0.26-2.1 mM, externally applied for 5-12 min) irreversibly decreased tau(-1), the rate constant of deactivation. At a pulse potential of 120 mV x tau(-1) decreased on average by 60%. The effect can be described as a 25 mV shift of the tau(-1)(V) curve. The activation curve and the curve relating steady-state current to pulse potential were shifted by similar amounts. The decrease of tau(-1) was the same at 0.26 and at 2.1 mM, but developed faster at the higher concentration. The slowing of deactivation was only seen when the cells were held at a potential of -20 mV. At this potential it developed with a time constant of 47 s. At more negative holding potentials (-40 or -70 mV) only a slight reduction of the peak occurred. The observations suggest preferential binding of DEPC to the open and inactivated channel states. The DEPC effect can possibly be explained by the reaction of DEPC with histidine residues or other amino acids in the external loops of the channel. However, dichloro-(2,2':6',2" terpyridine)-platinum (II) dihydrate (DTPD), another histidine-specific reagent, markedly decreased the peak current without affecting tau(-1). Therefore, the possibility that (some of) the effects of DEPC and DTPD are unrelated to their property as histidine-specific reagents cannot be excluded. PMID- 11489459 TI - Long-term treatment with antidepressant drugs reduces the sensitivity of cortical cholinergic neurons to the activating actions of stress and the anxiogenic drug FG 7142. AB - Certain antidepressant drugs exert an anxiolytic action in both humans and rodents. The effects of long-term treatment with imipramine or mirtazapine, two antidepressant drugs with different mechanisms of action, on the response of cortical cholinergic neurons to foot-shock stress or to the anxiogenic drug FG 7142 were investigated in freely moving rats. Chronic treatment with imipramine or mirtazapine reduced the increase in cortical acetylcholine output induced by foot-shock stress by approximately 50%. The same treatment also reduced the sensitivity of cortical cholinergic neurons to the stimulatory effect of acute administration of FG 7142. In contrast, the administration of a single dose of either antidepressant 40 min before foot shock or FG 7142 injection failed to increase the threshold of excitability of cortical cholinergic neurons. These results demonstrate that long-term treatment with either imipramine or mirtazapine reduces the sensitivity of cortical cholinergic neurons to stress or to an anxiogenic drug with an efficacy similar to that of acute administration of benzodiazepines. The neurochemical mechanism responsible for regulation of cholinergic neuron sensitivity might contribute to the modulation of cognitive function associated with emotional and affective disorders. PMID- 11489460 TI - The anthelmintic pyrantel acts as a low efficacious agonist and an open-channel blocker of mammalian acetylcholine receptors. AB - Pyrantel is an anthelmintic which acts as an agonist of nicotinic receptors (AChRs) of nematodes and exerts its therapeutic effects by depolarizing their muscle membranes. Here we explore at the single-channel level the action of pyrantel at mammalian muscle AChR. AChR currents are elicited by pyrantel. However, openings do not appear in clearly identifiable clusters over a range of pyrantel concentrations (1-300 microM). The mean open time decreases as a function of concentration, indicating an additional open-channel block. Single channel recordings in the presence of high ACh concentrations and pyrantel demonstrate that the anthelmintic acts as a high-affinity open-channel blocker. When analyzed in terms of a sequential blocking scheme, the calculated forward rate constant for the blocking process is 8x10(7) M(-1) x s(-1), the apparent dissociation constant is 8 microM at a membrane potential of -70 mV and the process is voltage dependent. Pyrantel displaces alpha-bungarotoxin binding but the concentration dependence of equilibrium binding is shifted towards higher concentrations with respect to that of ACh binding. Thus, by acting at the binding site pyrantel activates mammalian AChRs with low efficacy, and by sterical blockade of the pore, the activated channels are then rapidly inhibited. PMID- 11489461 TI - Substance P endopeptidase-like activity is altered in various regions of the rat central nervous system during morphine tolerance and withdrawal. AB - In this study the level of a substance P endopeptidase (SPE)-like activity was measured in different regions of the rat central nervous system (CNS) after chronic administration of morphine. Male rats (200-220 g) were randomly divided into four groups. Two groups were injected (s.c.) with morphine (10 mg/kg) twice daily, whereas the other two received saline under identical conditions. After 8 days, when animals were completely tolerant to morphine, one of the morphine treated groups and one group of saline-injected rats were given naloxone (s.c. 2 mg/kg). Withdrawal signs were observed and recorded. The enzyme activity was measured in extracts of the various CNS tissues by following the conversion of synthetic substance P (SP) to its N-terminal fragment SP(1-7) using a radioimmunoassay detecting this product. In discrete CNS areas including periaqueductal grey, spinal cord, substantia nigra and ventral tegmental area (VTA) a significant increase in enzyme activity was observed in the withdrawal group, while tolerant rats exhibited decreased SPE-like activity in the striatum (see Table 1). The enhanced enzyme activity during withdrawal is in agreement with our previous observation that the levels of SP(1-7) in rat brain are affected following naloxone precipitated withdrawal. In some tissues, including VTA, a correlation between the SPE-like activity and the intensity of the opioid abstinence was observed. Our result suggests that the elevated SPE-like activity is responsible for enhanced release of SP(1-7) in rats during morphine withdrawal, affirming a modulatory or regulative role of this enzyme in this state of opioid dependence. PMID- 11489462 TI - EP 91073 prevents EP 80661-induced penile erection: new evidence for the existence of specific EP peptide receptors mediating penile erection. AB - The effect of EP 91073, EP 51389, EP 70555 and EP 51216, peptide analogues of the growth hormone releasing peptide hexarelin, on penile erection induced by EP 80661 or EP 60761 injected into the paraventricular nucleus of the hypothalamus, was studied in male rats. Of the above peptides only EP 91073 (0.2-1 microg) was found capable of reducing penile erection induced by EP 80661 or EP 60761, when given into the paraventricular nucleus. Despite its ability to prevent EP peptide induced penile erection, EP 91073 (1 microg) was unable to prevent penile erection induced by the dopamine receptor agonist apomorphine (50 ng), oxytocin (30 ng) and N-methyl-D-aspartic acid (50 ng), when given into the paraventricular nucleus 10 min prior to the above substances. The EP 91073-induced prevention of penile erection occurred with a reduction in the increase in nitric oxide production that occurs in the paraventricular nucleus concomitant to penile erection induced by EP 80661 and EP 60761, as measured by intracerebral vertical microdialysis. The present results are in line with the hypothesis that EP 80661 and EP 60761 induce penile erection by activating specific receptors in the paraventricular nucleus, located possibly in oxytocinergic neurons mediating penile erection, and show that EP 91073 acts as an antagonist of these EP peptide receptors mediating penile erection. PMID- 11489463 TI - Enantio-selective cognitive and brain activation effects of N-ethyl-3,4 methylenedioxyamphetamine in humans. AB - In a randomised double-blind trial the subjective, neuropsychological and brain activation effects of the two enantiomers of the MDMA (ecstasy-) like drug N ethyl-3,4-methylenedioxyamphetamine (MDE) were studied in five normal subjects using functional magnetic resonance imaging (fMRI). (S)-MDE produced elevated mood, impairments in conceptually driven cognition and marked right frontal activation. In contrast, (R)-MDE produced increased depression, enhanced visual feature processing, and activation of visual cortical and left frontal areas. Plasma concentrations were higher for the (R)-enantiomer. The so-called entactogenic effects of MDE are likely to be caused by the (S)-enantiomer, whereas (R)-MDE appears to be responsible for neurotoxic effects. PMID- 11489464 TI - Tianeptine and its enantiomers: effects on spatial memory in rats with medial septum lesions. AB - Tianeptine, an atypical antidepressant that exhibits clinical efficacy in measures of depression and anxiety, has been reported to enhance learning and memory in rats under certain conditions, an effect not observed with other tricyclic antidepressants. The present study explores further the possibility that tianeptine or its enantiomers (S 16190 and S 16191) can enhance either learning or retention in animals in which the hippocampus has been made partially dysfunctional. The effects of tianeptine and its enantiomers were tested using an open field watermaze test, in rats with partial lesions of the medial septum/diagonal band of Broca (MSDB). When given to normal rats, tianeptine (10 mg/kg, i.p.) did not significantly affect learning as compared to animals injected with saline. We therefore created, in other animals, partial ibotenic acid lesions of MSDB and showed histochemically that these lesions reduced but did not abolish the density of acetylcholinesterase staining in the hippocampus. They impaired both the acquisition of place-navigation and the long-term retention of spatial information over 7 days. Against the baseline of impaired performance in animals with these lesions, neither tianeptine (10 mg/kg) nor its enantiomers (5 mg/kg) affected the rate of acquisition of place navigation. However, tianeptine did enhance the retention of spatial memory over 7 days. These results are discussed in relation to different effects that tianeptine may have on learning including its ability to block stress-induced dendritic re modelling of the hippocampus. PMID- 11489465 TI - Pharmacological comparison of human homomeric 5-HT3A receptors versus heteromeric 5-HT3A/3B receptors. AB - The present study determined the detailed pharmacological profile of heterologously expressed human (h) homomeric 5-HT3A receptors in direct comparison to heteromeric h5-HT3A/3B receptors. The very minor differences in their respective pharmacological profiles indicates that the 5-HT3B receptor subunit alters, predominantly, the biophysical rather than the pharmacological properties of the 5-HT3 receptor. PMID- 11489466 TI - Chromosome 7 monosomy and deletions in myeloproliferative diseases. AB - We studied deletion and monosomy of chromosome 7 in 150 patients with myeloproliferative diseases. We found 8/150 patients with monosomy 7 by cytogenetics and 4/150 with deletions of the long arm of chromosome 7 by restriction fragment length polymorphism (RFLP) analysis performed with Southern and polymerase chain reaction. To overcome limitation of RFLP analysis, we restricted loss of heterozygosity study with microsatellites to 45 patients, observing deletion 7q31.1 in 7/45 patients. In all patients with molecular alterations the deletion was observed only in myeloid cells, while the monosomy was detected in both myeloid precursor and lymphocytes. This finding suggests a CD34-totipotent stem cell origin for the monosomy and a colony forming unit - granulocyte, erythrocyte, monocyte, megakaryocytes (CFU-GEMM) stem cell origin for the deletions. PMID- 11489467 TI - Dysplastic versus proliferative CMML--a retrospective analysis of 91 patients from a single institution. AB - In chronic myelomonocytic leukemia (CMML) segregation of two subtypes has been suggested depending on WBC count-myelodysplastic (MD-CMML) and myeloproliferative (MP-CMML). In a retrospective analysis of 91 (60/31) previously untreated CMML patients, we compared the presenting clinical, haematological, laboratory and bone marrow features and examined the clinical impact of this reclassification. LDH values and bone marrow cellularity were significantly increased in MP-CMML. Median survival was significantly longer for patients with MD-CMML, progression rate was higher for MP-CMML. Patients with MD-CMML had longer median preleukemic duration; after transition to AML, MP-CMML patients had longer median survival. In MDS phase anemia was more common in MP-CMML and thrombocytopenia more common in MD-CMML whereas transfusion rates showed no difference. Evaluation of prognostic scoring systems for both groups confirmed that patients' characteristics and outcome could be well compared. Our data suggest that segregation into MD-CMML and MP-CMML is justified. PMID- 11489468 TI - Molecular detection of tumor cells at diagnosis invading the bone marrow and peripheral blood of patients with aggressive or indolent lymphomas. AB - We studied tumor cell invasions of bone marrow and peripheral blood in patients with various types of advanced non-Hodgkin's lymphoma by amplifying complementarity determining region III using the polymerase chain reaction (PCR) method and developing patient-specific probes. After molecular engineering, we could detect tumor cells in bone marrow from seven of 11 cases and in peripheral blood from six of 11 cases, despite negative results in four cases studied morphologically. Indolent cases were more likely to yield positive results than aggressive cases. The reason may be different biological behaviors among the histological types. PMID- 11489469 TI - AC133 antigen as a prognostic factor in acute leukemia. AB - AC133 is a novel 5-transmembrane antigen present on a CD34((bright)) subset of human hematopoietic stem cells (HSCs) and it is also expressed on the subset of CD34 positive (CD34(+)) leukemias. But the clinical significance of AC133 expression on leukemic blasts is not yet known. We investigated the expression of AC133 antigen on blast cells of acute leukemia. Forty-one cases of acute leukemia were examined for expression of AC133, CD34, and other antigens using multicolor flow-cytometry. Samples were considered positive if at least 20% of the cells specifically stained with monoclonal antibodies (MoAbs) revealed a higher fluorescence intensity compared to cells of corresponding negative control samples (=20% cut-off level). 14/36 (38.9%) acute myelogenous leukemia (AML) samples and 6/20 (30%) acute lymphoblastic leukemia (ALL) samples were positive for AC133, the difference was not significant. All AC133 positive (AC133(+)) leukemias expressed CD34, whereas 13 of 33 CD34(+) leukemias were negative for AC133, and AC133(+)/CD34(-) leukemia was not found. Expression rates of CD31, CD62L, CD62E, CD105 and CD144 were significantly higher in AC133(+) leukemia compared to those of AC133(-) leukemia (P=0.045, P<0.001, P<0.001, P<0.001, P=0.003, respectively), but bcl-2, CXCR-1, CXCR4, VLA-4, CD106 expression rates were not significantly different between AC133(+) and AC133(-) leukemias. None of the clinical prognostic markers such as age, hemogram, lactate dehydrogenase, and chromosomal aberration were significantly different between AC133(+) and AC133(-) leukemias. CR rates of AC133(+) AML and AC133(-) AML were not significantly different, although there was a trend toward higher CR rates in AC133(-) AML (18/22[81.8%] AC133(-) AML versus 9/14[64.3%] AC133(+) AML), but the 1-year relapse rate of AC133(+) AML was significantly higher than that of AC133(-) AML (8/9 (88.9%) versus 7/19 (36.8%), P=0.016). Median disease-free survival (DFS) times of AC133(+) and AC133(-) AML were significantly different (11 and 18 months, respectively, P=0.006), although overall survival (OS) times were not significantly different (AC133(+) 15 months versus AC133(-) 20 months, respectively, P=0.06). Similar results regarding clinical outcomes were found when AC133(+)/CD34(+) and AC133(-)/CD34(+) were analyzed separately, but the difference did not attain statistical significance. In ALL, 9/11 (81.8%) AC133(-) and 2/4 (50%) AC133(+) cases achieved CR, but the difference was not significant. Four of 11 AC133(-) ALL (36.4%) and 2 of 3 AC133(+) ALL (66.7%) relapsed within 1 year. In survival analysis, median DFS time and OS time of the AC133(+) group were 7 and 18 months, respectively, and these were not significantly different from those of the AC133(-) group (median DFS 15, OS 22 months, respectively). Our results demonstrate that AC133 expression in AML blasts is associated with poor clinical outcomes in terms of higher early relapse and shorter disease-free survival, suggesting that the AC133 antigen might provide the prognostic stratification of acute leukemia. However, to verify the effect of AC133 expression on the therapeutic outcomes of adult acute leukemia, further study including more cases is needed. PMID- 11489470 TI - Comparison of ethidium bromide-stained agarose gel electrophoresis and automated fragment analysis for evaluation of IgH gene products. AB - In acute lymphoblastic leukemia (ALL) patients, automated fluorescence fragment analysis (ALF) has been reported to improve the monoclonality detection rate of immunoglobulin heavy chain genes (IgH) polymerase chain reaction (PCR) analysis. This study performed complementary determining region (CDR) I and III PCR on samples from 135 patients with B-cell neoplasias and 25 healthy controls. The value of ALF was investigated in comparison to the widely used ethidium bromide (ETB)-stained agarose gels (AGGE). ETB-stained AGGE detected monoclonal CDR III PCR products in 53/72 ALL, in 22/34 non-Hodgkin's lymphoma (NHL), 13/22 multiple myeloma (MM), and 2/7 monoclonal gammopathies (MGUS). ALF identified monoclonal CDR III amplificates in 55/72 ALL, 23/34 B-NHL, 14/22 MM, and 2/7 MGUS. AGGE achieved clonal CDR I PCR results in 30/64 samples, while ALF detected 34 clonal CDR I product patterns. Taking together, ETB-stained AGGE revealed monoclonality in 120/199 PCR products versus 129/199 by ALF. Compared with AGGE and ETB staining, ALF offers a slightly increased sensitivity and can be recommended for the evaluation of difficult samples. PMID- 11489471 TI - HCV and tumors correlated with immune system: a case-control study in an area of hyperendemicity. AB - Hepatitis C virus (HCV) is a RNA virus that cannot be integrated with the host genome; it can, however, exert its oncogenetic potential indirectly by contributing to the modulator effects of the host immune system, probably through a capacity to elude the immune system. We have carried out a case controlled study on tumors correlated with the immune system (multiple myeloma, non-Hodgkin lymphoma and Hodgkin disease) and HCV, in a high prevalence area. The relationship between each cancer and HCV infection was assessed by means of odds ratios (ORs) and corresponding 95% confidence intervals. Risks were greater for B cell non-Hodgkin lymphoma (OR=3.7, 95%CI, 1.9-7.4, P=0.0001) and multiple myeloma (OR=4.5, 95%CI, 1.9-10.7, P=0.0004). Our study is particularly important for public health, since it shows that during the coming years in the South of Italy, because of the high prevalence of HCV, there are good reasons to expect not only an increase of liver cancer, but also an increased incidence of great number of tumors correlated with the immune system. PMID- 11489472 TI - Poor prognosis acute myelogenous leukemia: 3--biological and molecular biological changes during remission induction therapy. AB - This is the third paper in a series which describes a new remission induction regimen for patients with 'poor prognosis' acute myelogenous leukemia (AML). Twenty-four patients were treated with two one day pulses of chemotherapy separated by 96 h. Each pulse consisted of two doses of cytarabine and a single dose of mitoxantrone. Amifostine was administered three times a week after the second pulse of chemotherapy until treatment outcome became known. The first paper described the outcome of treatment while the second described the relationship of treatment outcome to the pretherapy characteristics of the leukemia. This paper describes the changes in the leukemia cells which occur during remission induction therapy. While only a limited number of specimens were available for each post treatment study, the studies demonstrated a profound fall in blood counts, BM cellularity, and telomerase activity in leukemia cells after pulse #1 of treatment. This fall was usually accompanied by a coordinate rise in IL6, TNFalpha, and IL1beta transcripts within the AML cells which survived chemotherapy. High levels of telomerase activity in the day 5 marrow was correlated with high levels of IL1beta transcripts which in turn were associated with treatment failure ascribable to resistant disease. PMID- 11489473 TI - Effects of Matrine on proliferation and differentiation in K-562 cells. AB - We investigated the effects of Matrine on proliferation by trypan blue exclusion and differentiation by benzidine staining positive cells in K-562 cells, assayed the telomerase activity using PCR-ELISA assay, analyzed cell cycle by fluorescence-activated cell sorter analysis of the DNA content, and also determined the gene expression level of c-myc, N-ras and p53 by northern blot and dot blot analysis. The results showed that with the addition of 0.1 mg/ml Matrine, cell growth was inhibited significantly by 4 days, benizidine-positive cells rose from 1% to 2% in control cells to 15% in treated cells on day 5; treatment of K-562 cells with 0.1 mg/ml Matrine for 5 days resulted in a marked inhibition in telomerase activity, in a manner that correlated with the extent of differentiation; after exposure to Matrine for 72 h, 64.6% cells were arrested in the G1-phase of the cell cycle, the fraction of cells in S-phase had decreased from 56.9% in control cells to 24.4% in differentiated cells, and the levels of N ras and p53 mRNA were remarkably increased for 24 and 48 h, respectively, c-myc mRNA expression level declined for 24 h and was inhibited significantly for 48 h. Our study confirmed that Matrine plays a significant effect on the inhibition of proliferation cells and inducing differentiation in K-562 cells. PMID- 11489474 TI - Inhibition of PI3-kinase sensitises HL60 human leukaemia cells to both chemotherapeutic drug- and Fas-induced apoptosis by a JNK independent pathway. AB - Increasing resistance to chemotherapeutic regimes remains a serious problem in the treatment of acute myeloid leukaemia. We have shown that phosphatidylinositol (PI) 3-kinase inhibition significantly sensitises the AML derived cell line, HL60 to chemotherapeutic drug- and Fas-induced apoptosis. PI3-kinase inhibition significantly potentiates cytotoxic drug-induced c-jun N-terminal kinase (JNK) activation, reported to be a requirement for apoptosis. However, JNK inhibition does not enhance cell viability following treatment with drug and inhibitor. Furthermore, PI3-kinase inhibition significantly increases sensitivity to apoptosis mediated by an exogenous receptor agonist, again by a JNK independent mechanism. These results suggest that PI3-kinase inhibitors could be of significant therapeutic importance, lowering the threshold for apoptosis induced by both chemotherapy and cell-mediated immune response. PMID- 11489475 TI - Pyridinyl imidazole inhibitor SB203580 activates p44/42 mitogen-activated protein kinase and induces the differentiation of human myeloid leukemia cells. AB - Various inhibitors of protein kinases regulate the growth and differentiation of human leukemic cell lines. The pyridinyl imidazole inhibitor SB203580 has been widely used to elucidate the role of p38 kinase in a wide array of biological systems. In the present investigation, we found that SB203580 effectively induced the granulocytic differentiation of human promyelocytic HL-60 cells. In addition to morphological differentiation, it also induced NBT-reduction, lysozyme activity and growth-inhibition. It also induced the differentiation of human myeloid leukemia HT93 and ML-1 cells, but not of other cell lines, such as NB4, U937, THP-1, K562 and HEL. This differentiation was not associated with the inhibition of p38 kinase activity, but was closely associated with the activation of extracellular signal-regulated kinase. These results demonstrate a new activity for this drug. PMID- 11489476 TI - Inhibition of protein kinase C delta has negative effect on anchorage-independent growth of BCR-ABL-transformed Rat1 cells. AB - The BCR-ABL oncoprotein transmits transformation signals mainly through pathways involving Ras, Myc and PI3 kinase. Here we report that inhibition of protein kinase C (PKC) delta had negative influence on anchorage-independent growth of Rat1 cells transformed by BCR-ABL. The effect was observed with delta isoform specific inhibitor rottlerin, but not with Go6976 that inhibits only conventional isoforms. The kinase activity of delta isoform was found to be roughly two-fold higher in BCR-ABL-expressing Rat1 cells than that in mock. Although overexpression of wild type PKC delta did not enhance soft agar colony number by BCR-ABL-transformed Rat1 cells, that of dominant-negative delta isoform reduced it by approximately 40%. PMID- 11489478 TI - Scientific summation and future goals. PMID- 11489479 TI - Delivery of molecular medicine to solid tumors: lessons from in vivo imaging of gene expression and function. AB - Extraordinary advances in molecular medicine and biotechnology have led to the development of a vast number of anti-cancer therapeutic agents. To reach cancer cells in a tumor, a blood-borne therapeutic molecule, particle or cell must make its way into the blood vessels of the tumor and across the vessel wall into the interstitium and finally migrate through the interstitium. Unfortunately, tumors often develop in ways that hinder each of these steps. Our research goals are to analyze each of these steps experimentally and theoretically and then integrate the resulting information in a unified theoretical framework. This paradigm of analysis and synthesis has allowed us to obtain a better understanding of microcirculatory barriers in solid tumors and to develop novel strategies to exploit and/or to overcome these barriers for improved cancer detection and treatment. PMID- 11489480 TI - Tumor chronobiology. AB - Traditionally, drug delivery has meant getting a simple chemical absorbed predictably from the gut or from the site of injection. A second-generation drug delivery goal has been the perfection of continuous constant rate (zero-order) delivery of simple xenobiotic molecules or common hormones. Living organisms are not 'zero-order' in their requirement for or response to drugs. They are predictable resonating dynamic systems, which require different amounts of drug at predictably different times within the circadian cycle in order to maximize desired and minimize undesired drug effects. PMID- 11489481 TI - Determinants of drug delivery and transport to solid tumors. AB - This presentation addresses the barriers and determinants and the importance of drug-induced apoptosis in drug transport and delivery to organs and solid tumors. In particular, we examined the roles of interstitial space, drug removal by capillaries, tissue structure and tissue composition on drug distribution. Drug transport in bladder tissues is described by the distributed model which combined monodimensional Fickian diffusion and first order removal of drug by the perfusing blood. Microscopic evaluation of the spatial drug distribution in bladder, prostate and tongue indicates heterogeneous drug distribution with large and erratic concentration gradient. In general, drug distribution favors interstitial space and vasculature, with little penetration in muscles. Drug penetration into 3-dimensional solid tumors is typically 5- to 10-fold slower than in monolayer cultures. The transport of highly protein-bound drugs such as paclitaxel and doxorubicin in a solid tumor is retarded by a high tumor cell density and enhanced by drug-induced apoptosis. Accordingly, the delivery of a highly protein-bound drug to cells in a solid tumor is affected by its apoptotic effects and is therefore determined by the drug concentration and the treatment duration, i.e. treatment schedule. Under in vitro and in vivo conditions, the delivery of highly protein-bound drugs to tumor can be enhanced by using a pretreatment that induces apoptosis and reduction in cell density, and by using treatment schedules designed to take advantage of these drug-induced changes in tumor tissue composition. In conclusion, in addition to the usual processes involved in drug transport such as distribution through vascular space, transport across microvessel walls, and diffusion through interstitial space in tumor tissue, other factors including tissue structure and composition and alteration by drug-induced apoptosis are important determinants of drug distribution in organs and solid tumors. PMID- 11489482 TI - Mechanism of tumor-targeted delivery of macromolecular drugs, including the EPR effect in solid tumor and clinical overview of the prototype polymeric drug SMANCS. AB - This review article describes three aspects of polymeric drugs. The general mechanism of the EPR (enhanced permeability and retention) effect and factors involved in the effect are discussed, in view of the advantages of macromolecular therapeutics for cancer treatment, which are based on the highly selective EPR related delivery of drug to tumor. Also described are advantages of more general water-soluble polymeric drugs as primary anticancer agents, using SMANCS as an example. Last, SMANCS/Lipiodol is discussed with reference to the type of formulation for arterial injection with most pronounced tumor selective delivery, as well as its advantages, precautions, and side effects from the clinical standpoint. PMID- 11489483 TI - Biodegradable polymer implants to treat brain tumors. AB - We have developed a systematic approach for the discovery and evaluation of local treatment strategies for brain tumors using polymers. We demonstrated the feasibility of polymer-mediated drug delivery by using the standard chemotherapeutic agent 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and showed that local treatment of gliomas by this method is effective in animal models of intracranial tumors. This led to clinical trials for glioma patients, and subsequent approval of Gliadel [(3.8% BCNU): p(CPP:SA)] by the FDA and other worldwide regulatory agencies. Twenty-two additional clinical trials are currently underway evaluating other issues related to the BCNU polymer, such as dosage, combination with systemic treatments, and combination with various forms of radiation and resistance modifiers. These trials are a result of laboratory investigations using brain tumor models; based on these models, other research groups have initiated clinical trials with novel combinations of different drugs and new polymers for both intracranial tumors (5-fluorouracil delivered via poly(D-L lactide-co-glycolide) polymer) and for tumors outside the brain (paclitaxel in PPE microspheres for ovarian cancer). Since only 1/3 of patients with glioblastoma multiforme (GBM) are sensitive to BCNU, the need to search for additional drugs continues. Although we are attacking major resistance mechanisms, there still will be tumors that do not respond to BCNU therapy but are sensitive to agents with different mechanisms of action, such as taxanes, camptothecin, platinum drugs, and antiangiogenic agents. Thus, it is necessary to explore multiple single agents and ultimately to combine the most effective agents for the clinical treatment of GBM. Furthermore, multimodal approaches combining radiotherapy with microsphere delivery of cytokines and antiangiogenic agents have demonstrated encouraging results. PMID- 11489484 TI - Targeted delivery of antisense oligonucleotides in cancer. AB - Formulations of antisense oligonucleotides (asODNs) against c-myb or c-myc protooncogenes have been prepared by a new technique that sequesters cationic lipid in the interior of a lipid particle. This technique results in high loading efficiency for the asODNs, small particle size and good stability. When targeted against melanoma cells or neuroblastoma cells via anti-GD(2) coupled at the particle surface, increased cell binding to the cells could be demonstrated. Targeted formulations showed greater inhibition of cell proliferation compared to non-targeted formulations or free drug. Inhibition of cell proliferation was demonstrated to be due to down-regulation of c-myb or c-myc protein expression. The formulations have long-circulation times in vivo, and evaluation for in vivo antitumor activity is currently underway. PMID- 11489485 TI - Retargeting of viral vectors to the folate receptor endocytic pathway. AB - Viral vectors with high transfection efficiencies are not always those with optimal target cell binding specificities. As a consequence, virus pseudotyping has been developed to endow transfection competent viruses with improved cell binding specificities and affinities. We have hypothesized that chemical conjugation of a virus to a cell specific ligand might also alter its target cell specificity and produce a virus that would transfect only the desired cell type. To test this concept, an ecotropic replication-defective myeloproliferative sarcoma retrovirus and an amphotropic murine adenovirus containing the gene for beta-galactosidase were chemically derivatized with folic acid. As expected from its strong ecotropism, the unmodified retrovirus did not induce beta galactosidase expression in nonhost KB cells, while the amphotropic adenovirus yielded high levels of gene expression in the same cell line. Surprisingly, although folate derivatization enabled avid binding of both viruses to folate receptor expressing KB cells, the folate conjugation did not promote retroviral gene expression and actually prevented the normal beta-galactosidase expression seen with the adenoviral vector. The fact that co-administration of excess free folic acid to block uptake by folate receptor-mediated endocytosis restored adenoviral gene expression to the level obtained with unmodified virus suggests that folate derivatization per se does not hamper viral activity. We, therefore, conclude that neither retroviral nor adenoviral delivery via the folate endocytosis pathway is compatible with viral gene expression in KB cells. PMID- 11489486 TI - Preparation and characterization of size-controlled polymeric micelle containing cis-dichlorodiammineplatinum(II) in the core. AB - Polymeric micelles of varying size in the range of 20 to 100 nm entrapping an antitumor drug, cis-dichlorodiammineplatinum(II) (cisplatin, CDDP), were prepared through the polymer-metal complex formation of CDDP with a mixture of poly(ethylene glycol)-poly(alpha,beta-aspartic acid) block copolymer (PEG-P(Asp)) and poly(alpha,beta-aspartic acid) homopolymer (P(Asp)) with the different feed ratio in distilled water. An increased ratio of P(Asp) to PEG-P(Asp) led to an increase in the micellar size in a controllable manner as well as prolongation in the induction period of the micellar decay accompanied by a sustained release of CDDP in physiological saline at 37 degrees C. All of the CDDP-loaded micelles with a different incorporation ratio of P(Asp) exhibited appreciable in vitro cytotoxicity due to CDDP release from the micelles by prolonged incubation. These CDDP-loaded micelles are expected to have potential utility in tumor-directed delivery system of CDDP through the modulated in vivo biodisposition based on the EPR effect. PMID- 11489488 TI - Tumor-targeted p53-gene therapy enhances the efficacy of conventional chemo/radiotherapy. AB - A long-standing goal in gene therapy for cancer is a stable, low toxic, systemic gene delivery system that selectively targets tumor cells, including metastatic disease. Progress has been made toward developing non-viral, pharmaceutical formulations of genes for in vivo human therapy, particularly cationic liposome mediated gene transfer systems. Ligand-directed tumor targeting of cationic liposome-DNA complexes (lipoplexes) is showing promise for targeted gene delivery and systemic gene therapy. Lipoplexes directed by ligands such as folate, transferrin or anti-transferrin receptor scFv, showed tumor-targeted gene delivery and expression in human breast, prostate, head and neck cancers. The two elements, ligand/receptor and liposome composition, work together to realize the goal of functional tumor targeting of gene therapeutics. The tumor suppressor gene, p53, has been shown to be involved in the control of DNA damage-induced apoptosis. Loss or malfunction of this p53-mediated apoptotic pathway has been proposed as one mechanism by which tumors become resistant to chemotherapy or radiation. The systemically delivered ligand-liposome-p53 gene therapeutics resulted in efficient expression of functional wild-type p53, sensitizing the tumors to chemotherapy and radiotherapy. This is a novel strategy combining current molecular medicine with conventional chemotherapy and radiotherapy for the treatment of cancer. The systemic delivery of normal tumor suppressor gene p53 by a non-viral, tumor-targeted delivery system as a new therapeutic intervention has the potential to critically impact the clinical management of cancer. PMID- 11489487 TI - Tumor targeting using anti-her2 immunoliposomes. AB - We have generated anti-HER2 (ErbB2) immunoliposomes (ILs), consisting of long circulating liposomes linked to anti-HER2 monoclonal antibody (MAb) fragments, to provide targeted drug delivery to HER2-overexpressing cells. Immunoliposomes were constructed using a modular strategy in which components were optimized for internalization and intracellular drug delivery. Parameters included choice of antibody construct, antibody density, antibody conjugation procedure, and choice of liposome construct. Anti-HER2 immunoliposomes bound efficiently to and internalized in HER2-overexpressing cells in vitro as determined by fluorescence microscopy, electron microscopy, and quantitative analysis of fluorescent probe delivery. Delivery via ILs in HER2-overexpressing cells yielded drug uptake that was up to 700-fold greater than with non-targeted sterically stabilized liposomes. In vivo, anti-HER2 ILs showed extremely long circulation as stable constructs in normal adult rats after a single i.v. dose, with pharmacokinetics that were indistinguishable from sterically stabilized liposomes. Repeat administrations revealed no increase in clearance, further confirming that ILs retain the long circulation and non-immunogenicity of sterically stabilized liposomes. In five different HER2-overexpressing xenograft models, anti-HER2 ILs loaded with doxorubicin (dox) showed potent anticancer activity, including tumor inhibition, regressions, and cures (pathologic complete responses). ILs were significantly superior vs. all other treatment conditions tested: free dox, liposomal dox, free MAb (trastuzumab), and combinations of dox+MAb or liposomal dox+MAb. For example, ILs produced significantly superior antitumor effects vs. non-targeted liposomes (P values from <0.0001 to 0.04 in eight separate experiments). In a non-HER2-overexpressing xenograft model (MCF7), ILs and non targeted liposomal dox produced equivalent antitumor effects. Detailed studies of tumor localization indicated a novel mechanism of drug delivery for anti-HER2 ILs. Immunotargeting did not increase tumor tissue levels of ILs vs. liposomes, as both achieved very high tumor localization (7.0-8.5% of injected dose/g tissue) in xenograft tumors. However, histologic studies using colloidal-gold labeled ILs demonstrated efficient intracellular delivery in tumor cells, while non-targeted liposomes accumulated within stroma, either extracellularly or within macrophages. In the MCF7 xenograft model lacking HER2-overexpression, no difference in tumor cell uptake was seen, with both ILs and non-targeted liposomes accumulating within stroma. Thus, anti-HER2 ILs, but not non-targeted liposomes, achieve intracellular drug delivery via receptor-mediated endocytosis, and this mechanism is associated with superior antitumor activity. Based on these results, anti-HER2 immunoliposomes have been developed toward clinical trials. Reengineering of construct design for clinical use has been achieved, including: new anti-HER2 scFv F5 generated by screening of a phage antibody library for internalizing anti-HER2 phage antibodies; modifications of the scFv expression construct to support large scale production and clinical use; and development of methods for large-scale conjugation of antibody fragments with liposomes. We developed a scalable two-step protocol for linkage of scFv to preformed and drug loaded liposomes. Our final, optimized anti-HER2 ILs-dox construct consists of F5 conjugated to derivatized PEG-PE linker and incorporated into commercially available liposomal doxorubicin (Doxil). Finally, further studies of the mechanism of action of anti-HER2 ILs-dox suggest that this strategy may provide optimal delivery of anthracycline-based chemotherapy to HER2-overexpressing cancer cells in the clinic, while circumventing the cardiotoxicity associated with trastuzumab+anthracycline. We conclude that anti-HER2 immunoliposomes represent a promising technology for tumor-targeted drug delivery, and that this strategy may also be applicable to other receptor targets and/or using other delivered agents. PMID- 11489489 TI - VIP receptors as molecular targets of breast cancer: implications for targeted imaging and drug delivery. AB - Receptors for vasoactive intestinal peptide (VIP-R) are overexpressed in human breast cancer. This phenomenon may have important diagnostic and therapeutic implications because carrier systems loaded with imaging or therapeutic agents, and with surface ligands to VIP-R could potentially be actively targeted to breast cancer. Previously, we have prepared sterically stabilized liposomes (SSL) with VIP non-covalently associated on their surface. However, these liposomes were not able to actively target to breast cancer in rats in situ, most probably due to dissociation of non-covalently associated VIP from SSL. Hence, there is a need to conjugate VIP covalently to SSL. This study aims to begin to address this issue and to test the targeting ability of VIP-SSL to n-methyl nitrosourea (MNU) induced rat breast cancer in vitro. First, VIP was conjugated to DSPE-PEG(3400) NHS [1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-n-[poly(ethylene glycol)]-N hydroxy succinamide, PEG M(w) 3400] under mild conditions to obtain a predominantly 1:1 conjugate of VIP and DSPE-PEG(3400) (DSPE-PEG(3400)-VIP), as evidenced by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). Next, DSPE-PEG(3400)-VIP was inserted into preformed fluorescent cholesterol (BODIPY-Chol) labeled SSL by incubation at 37 degrees C. To test breast cancer targeting ability in vitro, these VIP-SSL were subsequently incubated with MNU-induced rat breast cancer tissue sections. The results showed that when compared to fluorescent SSL without VIP or non-covalently attached VIP, significantly more VIP-SSL were attached to rat breast cancer tissues indicating that SSL with covalently attached VIP can be actively targeted to rat breast cancer tissues. This targeted carrier system is currently being explored for functional imaging and targeted chemotherapy of breast cancer. PMID- 11489490 TI - Polymer-drug conjugates, PDEPT and PELT: basic principles for design and transfer from the laboratory to clinic. AB - There are now at least seven polymer-drug conjugates that have entered phase I/II clinical trial as anticancer agents. These include N-(2 hydroxypropyl)methacrylamide (HPMA) copolymer-doxorubicin (PK1, FCE28068), HPMA copolymer-paclitaxel (PNU 166945), HPMA copolymer-camptothecin, PEG-camptothecin, polyglutamic acid-paclitaxel, an HPMA copolymer-platinate (AP5280) and also an HPMA copolymer-doxorubicin conjugate bearing additionally galactosamine (PK2, FCE28069). The galactosamine is used as a means to target the conjugate to liver for the treatment of primary and secondary liver cancer. Promising early clinical results with lysosomotropic conjugates has stimulated significant interest in this field. Ongoing research is developing (1) conjugates containing drugs that could otherwise not progress due to poor solubility or uncontrollable toxicity; (2) conjugates of agents directed against novel targets; and (3) two-step combinations such as polymer-directed enzyme prodrug therapy (PDEPT) and polymer enzyme liposome therapy (PELT) that can cause explosive liberation of drug from either polymeric prodrugs or liposomes within the tumour interstitium. Moreover, bioresponsive polymer-based constructs able to promote endosomal escape and thus intracytoplasmic delivery of macromolecular drugs (peptides, proteins and oligonucleotides) are also under study. PMID- 11489491 TI - Water soluble polymers in tumor targeted delivery. AB - The rationales for the use of water soluble polymers for anticancer drug delivery include: the potential to overcome some forms of multidrug resistance, preferential accumulation in solid tumors due to enhanced permeability and retention (EPR) effect, biorecognizability, and targetability. The utility of a novel paradigm for the treatment of ovarian carcinoma in an experimental animal model, which combines chemotherapy and photodynamic therapy with polymer-bound anticancer drugs is explained. Research and clinical applications as well as directions for the future development of macromolecular therapeutics are discussed. PMID- 11489492 TI - PEG drugs: an overview. AB - No low molecular weight (<20000) poly(ethylene glycol) (PEG) small molecule drug conjugates, prepared over a 20-year period, have led to a clinically approved product. In this area, published studies for these types of compounds have been scrutinized and their properties compared and contrasted to higher molecular weight conjugates where, during the past 5 years, a renaissance in the field of PEG (anticancer) drug conjugates has taken place. This new development has been attributed to the use of higher molecular weight PEGs (>20000), and especially employing PEG 40000 which is estimated to have a plasma circulating half life of approximately 8-9 h in the mouse. This recent resuscitation of small organic molecule delivery by high molecular weight PEG conjugates was founded on meaningful in vivo testing using established tumor models, and has led to a clinical candidate. Recent applications of high molecular weight PEG prodrug strategies to amino containing drugs are also detailed, and potential applications to proteins are proposed. PMID- 11489493 TI - VEGF-VEGF receptor complexes as markers of tumor vascular endothelium. AB - Vascular endothelial growth factor (VEGF) is a primary stimulant of the vascularization of solid tumors and has therefore been the focus of intense research aimed at blocking its activity in solid tumors. VEGF production by tumor cells is induced by oncogenic gene mutations and hypoxic conditions inside the tumor mass. VEGF receptor expression on endothelial cells lining blood vessels in the tumor is also induced by hypoxia and the increased local concentration of VEGF. Therefore in the tumor microenvironment there is an upregulation of both VEGF and its receptor leading to a high concentration of occupied receptor on tumor vascular endothelium. The VEGF-VEGF receptor complex (VEGF-VEGFR) presents an attractive target for the specific delivery of drugs or other effectors to tumor endothelium. Herein we review the development of monoclonal antibodies that selectively bind to the VEGF-VEGFR and their use as targeting agents that selectively bind to VEGF activated blood vessels. Additionally, we summarize the properties of 2C3, a novel monoclonal anti-VEGF antibody that blocks VEGF from binding to VEGFR2 but not VEGFR1. 2C3 may be utilized as both an anti-angiogenic agent by inhibiting VEGFR2 activity and potentially as a vascular targeting agent by binding to blood vessels that express the VEGF-VEGFR1 complex. PMID- 11489494 TI - Targeting drug delivery to radiation-induced neoantigens in tumor microvasculature. AB - Radiation can be used to guide drugs to specific sites such as neoplasms or aberrant blood vessels. When blood vessels are treated with ionizing radiation, they respond by expressing a number of cell adhesion molecules and receptors that participate in homeostasis. Examples of radiation-induced molecules in blood vessels include ICAM-1, E-selectin, P-selectin and the beta(3) integrin. We have observed that the endothelium and blood components respond to oxidative stress in a similar, if not identical manner in all tumor models. Although we have identified several other radiation-induced molecules within tumor blood vessels, the beta(3) target for drug delivery achieves the greatest site-specific peptide binding within irradiated tumor blood vessels. We have focused on peptides and antibodies that bind to integrin beta(3). beta(3)-binding proteins have been conjugated to fluorochromes and radionuclides to study the site specificity and microscopic distribution. We have found immunofluorescent and immunohistochemical staining of beta(3) within the lumen of blood vessels immediately following irradiation. To determine whether it is feasible to guide drug delivery to irradiated tumors, we studied ligands to alpha(2b)beta(3) (fibrinogen). Peptides within fibrinogen that bind to alpha(2b)beta(3) includes the dodecapeptide, HHLGGAKQAGDV and the RGD peptide. We utilized 131I conjugation to these ligands to study the biodistribution in tumor bearing mice. Our clinical trial consists of the RGD peptidomimetic, biapcitide, labeled with 99mTc. This study shows that it is feasible to guide drugs to human neoplasms by use of radiation-guided peptides. These studies have shown that peptides that bind to these integrins bind to tumors following exposure to ionizing radiation. PMID- 11489495 TI - Finite element model of antibody penetration in a prevascular tumor nodule embedded in normal tissue. AB - We have developed a pharmacokinetic model for monoclonal antibodies (mAb) to aid in investigating protocols for targeting small primary tumors or sites of metastatic disease. The model describes the uptake of systemically-administered antibody by a prevascular spherical tumor nodule embedded in normal tissue. The model incorporates plasma kinetics, transcapillary transport, interstitial diffusion, binding reactions, and lymphatic clearance. Antigen internalization can easily be incorporated. Simulations obtained from a three-dimensional finite element analysis are used to assess errors in predictions from earlier models in which the influence of the normal tissue was collapsed into a boundary condition at the tumor surface. The model employing a Dirichlet boundary condition substantially overpredicted the mean total tumor mAb concentration at all times. Although the model with a concentration-dependent flux (composite) boundary condition underpredicted mAb concentration, the discrepancy with finite element results is only notable at early times. Sensitivity analyses were performed on mAb dose and on the coefficients for mAb diffusion in the tissue regions, since reported antibody diffusivity values have varied over 30-fold. The results of the study suggest that mAb diffusivity and mAb binding site density in tumors should have major influences on optimizing doses and scheduling of mAb administration in tumor targeting protocols. PMID- 11489496 TI - Synthesis and characterization of HPMA copolymer-aminopropylgeldanamycin conjugates. AB - Geldanamycin (GDM) is a benzoquinone ansamycin antibiotic with anticancer activity. The use of drug delivery systems based on N-(2 hydroxypropyl)methacrylamide (HPMA) copolymers containing lysosomally degradable oligopeptide (GFLG) spacers results in an increased therapeutic efficacy of anticancer drugs. The objective of this study was to synthesize HPMA copolymer GDM conjugates with anticancer activity and reduced toxic side-effect of the compound. 17-(3-Aminopropylamino)-17-demethoxygeldanamycin (AP-GDM) was synthesized and converted into a polymerizable GDM derivative, N methacryloylglycylphenylalanylglycyl-17-(3-aminopropylamino)-17 demethoxygeldanamycin [MA-GFLG-(AP-GDM)]. The structures of AP-GDM and MA-GFLG (AP-GDM) were validated by mass spectroscopy, elemental analysis, and two dimensional nuclear magnetic resonance. MA-GFLG-(AP-GDM) was copolymerized with HPMA and N-methacryloyglycylglycine p-nitrophenylester by radical precipitation polymerization. Water-soluble HPMA copolymer-AP-GDM conjugates (M(r)=16 kDa) were obtained. Monoclonal antibody OV-TL16, which recognizes the OA-3 antigen expressed on the OVCAR-3 human ovarian carcinoma cell line, was optionally attached to the HPMA copolymer-AP-GDM conjugate. Cytotoxicity of polymer-bound AP GDM (both targeted and non-targeted) was determined using OVCAR-3 and another human ovarian carcinoma cell line, A2780. The HPMA copolymer-AP-GDM conjugate was cytotoxic toward A2780 cells. Attachment of OV-TL16 antibody enhanced cytotoxicity of the conjugate toward OVCAR-3 cells. PMID- 11489497 TI - Drug targeting using thermally responsive polymers and local hyperthermia. AB - We report a new thermal targeting method in which a thermally responsive drug carrier selectively accumulates in a solid tumor that is maintained above physiological temperature by externally applied, focused hyperthermia. We synthesized two thermally responsive polymers that were designed to exhibit a lower critical solution temperature (LCST) transition slightly above physiological temperature: (1) a genetically engineered elastin-like polypeptide (ELP) and (2) a copolymer of N-isopropylacrylamide (NIPAAm) and acrylamide (AAm). The delivery of systemically injected polymer-rhodamine conjugates to solid tumors was investigated by in vivo fluorescence video microscopy of ovarian tumors implanted in dorsal skin fold window chambers in nude mice, with and without local hyperthermia. When tumors were heated to 42 degrees C, the accumulation of a thermally responsive ELP with a LCST of 40 degrees C was approximately twofold greater than the concentration of the same polymer in tumors that were not heated. Similar results were also obtained for a thermally responsive poly(NIPAAM-co-AAm), though the enhanced accumulation of this carrier in heated tumors was lower than that observed for the thermally responsive ELP. These results suggest that enhanced delivery of drugs to solid tumors can be achieved by conjugation to thermally responsive polymers combined with local heating of tumors. PMID- 11489498 TI - Doxorubicin bound to a HPMA copolymer carrier through hydrazone bond is effective also in a cancer cell line with a limited content of lysosomes. AB - We have synthesized conjugates containing doxorubicin (DOX) bound to oligopeptide side chains (GlyGly or GlyPheLeuGly) of a water-soluble copolymer carrier based on poly[N-(2-hydroxypropyl)methacrylamide] (PHPMA) either through proteolytically (PK1 conjugates) [Synthetic polymeric drugs. U.S. Patent 5,037,883 (1991)] or hydrolytically cleavable bond (HC conjugates). Pharmacological efficacy of PK1 and HC conjugates was compared in vitro on murine: T-cell lymphoma EL4, B-cell leukemia BCL1, B-cell lymphoma 38C13, leukemia P388 and Con A-stimulated A/Ph splenocytes and on human: primary (SW480) and metastatic (SW620) colorectal cancer cell lines parent and transfected with Thy 1.2 gene [2] and on erythromyeloid leukemia cell line K 562. Inhibition of proliferation determined by 3[H]-thymidine incorporation revealed that the cytostatic effect of HC conjugates is up to two orders of magnitude higher compared to PK1 conjugates. In some cancer cell lines (SW 620/T, SW 480) the pharmacological activity of HC conjugates is in vitro comparable with the activity of the free drug. Unlike PK1 conjugates, HC conjugates with a lysosomally degradable spacer (GlyPheLeuGly) are less effective compared to HC conjugates containing lysosomally non-degradable spacer (GlyGly). Moreover, HC conjugates exert pronounced anti-proliferative activity also in erythroblastoid leukemia cell line K 562 with a limited content of lysosomes. PMID- 11489499 TI - Tumor targeting by conjugation of DHA to paclitaxel. AB - Targeting an anti-cancer drug to tumors should increase the Area Under the drug concentration-time Curve (AUC) in tumors while decreasing the AUC in normal cells and should therefore increase the therapeutic index of that drug. Anti-tumor drugs typically have half-lives far shorter than the cell cycle transit times of most tumor cells. Tumor targeting, with concomitant long tumor exposure times, will increase the proportion of cells that move into cycle when the drug concentration is high, which should result in more tumor cell killing. In an effort to test that hypothesis, we conjugated a natural fatty acid, docosahexaenoic acid (DHA), through an ester bond to the paclitaxel 2'-oxygen. The resulting paclitaxel fatty acid conjugate (DHA-paclitaxel) does not assemble microtubules and is non-toxic. In the M109 mouse tumor model, DHA-paclitaxel is less toxic than paclitaxel and cures 10/10 tumored animals, whereas paclitaxel cures 0/10. One explanation for the conjugate's greater therapeutic index is that the fatty acid alters the pharmacokinetics of the drug to increase its AUC in tumors and decrease its AUC in normal cells. To test that possibility, we compared the pharmacokinetics of DHA-paclitaxel with paclitaxel in CD2F1 mice bearing approximately 125 mg sc M109 tumors. The mice were injected at zero time with a bolus of either DHA-paclitaxel or paclitaxel formulated in 10% cremophor/10% ethanol/80% saline. Animals were sacrificed as a function of time out to 14 days. Tumors and plasma were frozen and stored. The concentrations of paclitaxel and DHA-paclitaxel were analyzed by LC/MS/MS. The results show that DHA targets paclitaxel to tumors: tumor AUCs are 61-fold higher for DHA paclitaxel than for paclitaxel at equitoxic doses and eight-fold higher at equimolar doses. Likewise, at equi-toxic doses, the tumor AUCs of paclitaxel derived from i.v. DHA-paclitaxel are 6.1-fold higher than for paclitaxel derived from i.v. paclitaxel. The tumor concentration of paclitaxel derived from i.v. paclitaxel drops rapidly, so that by 16 h it has fallen to the same concentration (2.8 microM) as after an equi-toxic concentration of DHA-paclitaxel. In plasma, paclitaxel AUC after an MTD dose of DHA-paclitaxel is approximately 0.5% of DHA paclitaxel AUC. Thus, the increase in tumor AUC and the limited plasma AUC of paclitaxel following DHA-paclitaxel administration are consistent with the increase in therapeutic index of DHA-paclitaxel relative to paclitaxel in the M109 mouse tumor model. A phase I clinical study has been completed at The Johns Hopkins Hospital to evaluate the safety of DHA-paclitaxel in patients with a variety of solid tumors. Twenty-one patients have been treated to date. The recommended phase II dose is 1100 mg/m(2), which is equivalent to 4.6 times the maximum approved paclitaxel dose on a molar basis. No alopecia or significant peripheral neuropathy, nausea, or vomiting have been observed. Asymptomatic, transient neutropenia has been the primary side effect. Eleven of 22 evaluable phase I patients transitioned from progressive to stable disease, as assessed by follow-up CT. Significant quality of life improvements have been observed. Thus, DHA-paclitaxel is well tolerated in patients and cures tumors in mice by targeting drug to tumors. PMID- 11489500 TI - Potentiation of radioresponse by polymer-drug conjugates. AB - Although combined chemotherapy and radiotherapy has produced significantly improved response and survival rates among cancer patients, there is still a compelling need to establish the most effective way to deliver these agents. We hypothesize that the radiosensitizing effect of a chemotherapeutic agent can be further enhanced if the drug is delivered at an optimal concentration and is maintained in the tumor for a prolonged period. Using a water-soluble poly(L glutamic acid)-conjugated paclitaxel (PG-TXL) as a model compound, we investigated whether paclitaxel delivered by means of polymeric carrier could increase the tumor's response to radiation. Mice bearing 8-mm syngeneic ovarian carcinoma OCa-1 tumors implanted intramuscularly were treated with i.v. injected PG-TXL alone or in combination with single doses of local radiation. The enhancement factors at 24 h interval, as measured by incremental tumor growth delay compared with radiation alone, ranged from 2.48 to 4.28. The values varied as a function of radiation dose. The enhancement of radioresponse is also a function of time interval between injection of PG-TXL and tumor irradiation. The enhancement factor increased with decreasing interval, suggesting that radiation may in turn mediate the sensitivity of tumor toward PG-TXL. Thus, the mechanism of PG-TXL's radiopotentiation activity is probably multifactorial. Remarkably, while combined radiation and TXL produced additive or even sub-additive interaction when radiation preceded TXL injection, combined radiation and PG-TXL produced synergistic interaction in a mammary MCa-4 tumor model. Radiation significantly increased tumor uptake of PG-TXL, suggesting a potential role of radiation-modulated antitumor activity of polymeric drugs. Our data support a treatment strategy combining radiation and polymeric chemotherapy that may have important clinical implications in terms of scheduling and optimization of the therapeutic ratio. PMID- 11489501 TI - Water-soluble poly-(L-glutamic acid)-Gly-camptothecin conjugates enhance camptothecin stability and efficacy in vivo. AB - The therapeutic efficacy of 20(s)-camptothecin (CPT) is limited in humans by the instability of the active lactone form due to preferential binding of the carboxylate to serum albumin and by difficulty in formulation. Formation of an ester bond with an amino acid via the hydroxyl group at carbon 20 of CPT stabilizes the lactone. Linking CPT to a high molecular weight (MW) anionic polymer enhances solubility and improves distribution to the tumor through enhanced permeability and retention (EPR effect). Poly-(L-glutamic acid) (PG) is an anionic homo-polymer that can theoretically bind one molecule of a drug via the gamma carboxylic acid of each monomeric subunit. It has been used to make a water-soluble PG-paclitaxel conjugate currently in Phase II clinical trials that contains 37% paclitaxel by weight and is administered in a 10 min infusion without pre-medication. We evaluated the anti-tumor activity of PG conjugates of CPT after a single intraperitoneal injection using subcutaneous murine B-16 melanoma tumor growth as an indicator. Interposition of a glycine (gly) linker allowed CPT loading up to 50% w/w on the polymer. Increasing the PG MW from 33 to 49 kDa enhanced the efficacy without altering the maximum tolerated dose (MTD). In athymic mice bearing ectopic human colon or lung tumors, efficacy was enhanced compared to free camptothecin. Thus, as with paclitaxel, conjugation of CPT to PG enhanced pharmaceutical properties and preclinical efficacy. PMID- 11489502 TI - Combination chemotherapy and photodynamic therapy of targetable N-(2 hydroxypropyl)methacrylamide copolymer-doxorubicin/mesochlorin e(6)-OV-TL 16 antibody immunoconjugates. AB - The aim of this study was to evaluate the combination chemotherapy and photodynamic therapy of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-bound doxorubicin (DOX) and mesochlorin e(6) (Mce(6)) targeted with an OV-TL 16 monoclonal antibody (P-DOX-Ab and P-Mce(6)-Ab, respectively) in nude mice bearing human ovarian OVCAR-3 carcinoma xenografts. P-DOX-Ab and P-Mce(6)-Ab were synthesized by first conjugating DOX or Mce(6) to an HPMA copolymer precursor (Mw=21000), then reacting with OV-TL 16 antibody. The immunoconjugates were purified by size exclusion chromatography on Superose 6 column and analyzed. The Mce(6) concentration in tissues was determined by a fluorescence assay. Eighteen hours after administration, the tumors received a light dose of 220 J/cm(2) from a KTP 650-nm dye-laser. P-DOX-Ab and P-Mce(6)-Ab had polymer:drug:protein weight ratios of 32:3:62 and 26:2:72, corresponding to polymer:drug:protein molecular ratios of approximately 4:14:1 and 3:8:1, respectively. The biodistribution results indicated that the percentage of total administered dose of Mce(6) in tumors reached approximately 1% for the nontargeted conjugate at 18 h after administration, while that of P-Mce(6)-Ab was approximately 13 times higher. Nude mice bearing OVCAR-3 xenografts that received one i.v. dose of P-DOX-Ab (2.2 mg/kg DOX equivalent) and P-Mce(6)-Ab (1.5 mg/kg Mce(6) equivalent) with light irradiation achieved a xenograft cure rate of more than 60%. The incorporation of OV-TL 16 antibody dramatically enhanced the accumulation in tumors with a concomitant increase in the therapeutic efficacy of P-DOX-Ab and P-Mce(6)-Ab in combination therapy, which may probably be attributed to both antibody targeting and enhanced permeability and retention (EPR) effects. PMID- 11489503 TI - Targeting of acivicin prodrugs as antibody conjugates. AB - The ectopeptidase gamma-glutamyltranspeptidase (gamma-GT) is overexpressed in myeloid leukemias. Its specific inhibitor, acivicin, was previously shown to induce an inhibitory growth effect associated with an induction of morphological features characteristic of macrophage maturation. We have considered a construction in which an antibody linked to a prodrug of acivicin will target acivicin to tumoral cells. In a first set of experiments we have synthesized a chromogenic model of this prodrug to validate this concept of prodrug, allowing an amine function to be released upon esterase action. Thereafter this model was applied to acivicin. The acivicin prodrug is inactive toward purified gamma-GT, and recovers its inhibitory activity under the effect of esterase. PMID- 11489504 TI - Development of a novel molecular adapter for the optimization of immunotoxins. AB - Immunotoxins consisting of catalytic domains of natural toxins and tumor-specific ligands were modified by introducing a molecular adapter that is able to transport the toxic domain more efficiently into cells. The adapter is a three component structure: its core is a membrane transfer sequence (MTS) flanked by two different cleavable sequences. The directed and irreversible cellular uptake of the construct is driven by either enzymatic or chemical cleavage of the two flanking sequences. In our studies, the purified A-chain of diphtheria toxin (DT) was coupled to two different MTSs via disulfide bonds. A cytotoxicity assay revealed that the constructs containing the MTSs were more potent than DT A-chain alone and that the disulfide bond was cleaved. PMID- 11489505 TI - Preparation and biological evaluation of polymerizable antibody Fab' fragment targeted polymeric drug delivery system. AB - A new polymerizable antibody Fab' fragment with a PEG spacer (MA-PEG-Fab') was prepared from OV-TL 16 antibody, specific against the OA-3 antigen expressed on most human ovarian carcinomas. The MA-PEG-Fab' possessed a higher reactivity in the copolymerization with N-(2-hydroxypropyl)methacrylamide (HPMA) than the polymerizable Fab' fragment MA-Fab' with a short spacer. The MA-PEG-Fab' was copolymerized with HPMA and MA-Gly-Phe-Leu-Gly-Mce(6) producing an Fab' targeted HPMA copolymer-Mce(6) conjugate. The number and weight average molecular weights of the copolymer were 164000 and 271000 Da, respectively. About two MA-PEG-Fab' fragments per chain were incorporated in the copolymer conjugates. Preliminary in vivo antitumor studies indicated that the Fab' targeted conjugates showed a higher efficacy of tumor growth inhibition in nude mice than the non-targeted conjugate. PMID- 11489506 TI - The effects of subcellular localization of N-(2-hydroxypropyl)methacrylamide copolymer-Mce(6) conjugates in a human ovarian carcinoma. AB - Photosensitizers, light-sensitive compounds, become activated upon illumination with a specific wavelength of light generating cytotoxic oxygen species. Due to the short half-life of singlet oxygen, the subcellular site of localization and excitation affects the type of cellular damage produced as well as cellular responses to different types of photodamage created within the cell. Here, we investigated the effects of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer mesochlorin e(6) monoethylenediamine (Mce(6)) conjugates localized to different subcellular compartments. Temperature was utilized to achieve subcellular localization of conjugates and subcellular fractionation was performed to confirm localization patterns of HPMA copolymer-Mce(6) conjugates. Cytotoxicity studies suggest plasma membrane and late endosomes were more sensitive to photodamage than lysosomal compartments as observed by an approximate 2-fold decrease in the IC(50) compared to lysosomally accumulated conjugate. Releasing Mce(6) from the polymer backbone within lysosomal compartments significantly lowered the IC(50) when compared to HPMA copolymer conjugates with Mce6 bound via a nondegradable linkage. These differences will prove useful in the future design of HPMA copolymer-Mce(6) conjugates for the treatment of ovarian cancer. PMID- 11489507 TI - Water-soluble HPMA copolymer-wortmannin conjugate retains phosphoinositide 3 kinase inhibitory activity in vitro and in vivo. AB - Phosphoinositide kinases and ATM-related genes play a central role in many physiological processes. Activation of phosphoinositide 3-kinase (PI 3-kinase) is essential for signal transduction by many growth factors and oncogenes and may contribute to tumor progression. In the nanomolar range, Wortmannin (WM), a fungal metabolite, is a potent inhibitor of type I PI 3-kinase; it covalently modifies its catalytic subunit. Because WM is soluble only in organic solvents and unstable in water, there are difficulties in its use in vivo. To generate a water-soluble WM derivative, we used a conjugate of N-(2 hydroxypropyl)methacrylamide (HPMA) copolymer and 11-O-desacetylwortmannin (DAWM), which has a slightly lower inhibitory activity than WM. We covalently attached DAWM to HPMA copolymer containing oligopeptide (GFLG) side-chains. The final product had an estimated molecular mass of 20 kDa and contained 2 wt.% of DAWM. The HPMA copolymer (PHPMA)-DAWM conjugate inhibited type I PI 3-kinase activity in vitro and growth factor-stimulated activation of Akt in vivo; it possessed approximately 50% of the inhibitory activity of DMSO solubilized WM. The specificity and stability of the PHPMA-DAWM conjugate is currently under investigation. The new water-soluble form of WM may be useful in investigations of the role of PI 3-kinase in tumor progression and other cellular biological functions in vivo. PMID- 11489508 TI - Potential of lectin-N-(2-hydroxypropyl)methacrylamide copolymer-drug conjugates for the treatment of pre-cancerous conditions. AB - N-(2-Hydroxypropyl)methacrylamide (HPMA)-lectin (wheat germ agglutinin (WGA), peanut agglutinin (PNA)) drug conjugates for treatment of the pre-cancerous conditions ulcerative colitis and Barrett's esophagus are being developed. Cell surface glycoproteins that are altered in disease and development bind lectins. PNA binds alpha-lactose and the Thomsen-Friedenreich (TF) antigen, a disease- and development-associated glycoprotein. PNA incorporation in conjugates may allow for preferential delivery to diseased over healthy tissues. Conjugates were prepared by attaching lectins to HPMA copolymers via an amide linkage. Frontal affinity chromatography was used to measure dissociation constants (K(d)) of free and conjugated lectins. Animal models of colitis (DSS, TNBS/EtOH) were developed. Human biopsy specimens were obtained. Free and HPMA copolymer-conjugated FITC labeled lectin and anti-TF antigen antibody binding patterns were examined in normal neonatal, adult and diseased rodent tissues and normal and diseased human tissues. K(d) values of free and conjugated lectins were similar ( approximately 10(-5) M(-1)). Free and conjugated lectins had comparable binding patterns. In health, strong WGA binding was seen in goblet cells; PNA binding was minimal, occurring only in the supranuclear goblet cell region. In disease, WGA binding was not altered, but PNA binding was increased in both human and rodent tissues; entire goblets bound the lectin. Anti-TF antigen antibody binding was minimal, but did overlap with PNA binding patterns both in normal and diseased tissues. Conjugation of lectins to HPMA copolymers does not affect binding affinity. Alterations in glycoprotein structures in development and disease resulted in modified lectin binding patterns. In development and disease, the PNA binding seen was to the TF antigen and other lactose-containing glycoproteins. The results suggest that site-specific delivery of therapeutic agents such as cyclosporin A (CsA) for ulcerative colitis and mesochlorin e(6) for Barrett's esophagus may be achieved. P(HPMA)-lectin-CsA conjugates have been prepared and preliminary in vivo studies are underway. PMID- 11489509 TI - Development of the polymer micelle carrier system for doxorubicin. AB - We show the result of pre-clinical study of NK911, a polymeric micelle carrier system for doxorubicin (DOX). The NK911 micelle carrier consists of polyethyleneglycol and conjugated doxorubicin-polyaspartic acid. It has high hydrophobic inner core, and therefore can entrap the sufficient amount of DOX. NK911 has a small particle size of about 40 nm in diameter that accumulates in tumor tissue by EPR effect showing much stronger activity than the free DOX. We plan to perform a clinical trial at National Cancer Center Hospital, Japan from 2001. PMID- 11489510 TI - Potential of block copolymer- and immuno-conjugates for tumor-targeted delivery of Bowman-Birk soybean proteinase inhibitor. AB - The present work reports the effect of conjugation of the anticarcinogenic and antitumor soybean Bowman-Birk protease inhibitor (BBI) with amphiphilic block copolymer of ethylene oxide and propylene oxide (PEO-PPO) as well as with monoclonal antibody via clinical dextran (D) on tumor-targeted delivery of BBI. PMID- 11489511 TI - Sterically stabilized polyplex: ligand-mediated activity. AB - Synthetic vectors have been considered as a safer and more versatile alternative to viral-based gene delivery systems. A variety of very simple synthetic vector systems, e.g., cationic lipid- and polymer-complexed plasmid DNA have activity in vivo but it appears to be mediated by non-specific electrostatic interactions limiting targeting. In order to avoid these problems, we designed a sterically stabilized layered colloidal system. The steric polymer coating reduces non specific interactions. We have synthesized a PEG conjugate of PEI that complexes DNA to form small, stable colloids with a steric polymer coat on their surface. The polymer enhances colloidal stability and reduces non-specific binding and toxicity. It also renders the complex inactive presumably due to reduced binding. Ligands are then appended to the distal end of the steric polymer to restore cell binding and expression at target cells. We prepared conjugates with RGD peptide ligands appended to the distal end of the steric polymer. The resulting conjugates also form complexes but with ligands exposed on their surface restoring binding and activity. Labeled oligonucleotides and DNA were used to measure intracellular distribution. Oligonucleotides are found localized in the nucleus, whereas the labeled plasmid DNA remained in the cytoplasm. Import of plasmid DNA into the nucleus appears to be very inefficient yet sufficient for expression. PMID- 11489512 TI - Salmonella-based tumor-targeted cancer therapy: tumor amplified protein expression therapy (TAPET) for diagnostic imaging. AB - In preclinical studies, genetically engineered Salmonella have the ability to localize, selectively accumulate, and persist within transplantable murine tumors, spontaneous murine tumors and human tumor xenographs, and can express therapeutic proteins at high levels. These strains of engineered non-virulent Salmonella typhimurium display the capacity to accumulate and grow selectively in a variety of tumor types and to inhibit the growth of primary and metastatic tumors following intravenous injection into tumor-bearing mice. One strain of the bacteria (VNP20009) which has endogenous antitumor activity is currently in Phase I clinical trials. The bacteria are highly attenuated and genetically stable. The combination of the lipid mutation and the purine auxotrophy attenuate the virulence of the bacteria by greater than 10000-fold and enhance the specificity of the bacteria for tumor tissue. These bacteria have been found to be safe in mice, pigs and monkeys when administered intravenously. Second-generation Salmonella vectors will be developed to include transgenes that will express therapeutic agents and reporter transgenes for non-invasive imaging. We have performed a preliminary study to demonstrate localization of [(14)C]FIAU in tumored mice pretreated with Salmonella expressing HSV1-TK. The [(14)C]FIAU radioactivity and bacterial count data strongly support a Salmonella(TK) dependent [(14)C]FIAU accumulation of at least 30-fold higher in tumor tissue compared to muscle tissue. These data warrant further investigation on the use of genetically engineered Salmonella as a systemically administered tumor-specific agents for tumor therapy and delivery of diagnostic imaging markers. PMID- 11489513 TI - Tumour targeted delivery of encapsulated dextran-doxorubicin conjugate using chitosan nanoparticles as carrier. AB - Doxorubicin (DXR) commonly used in cancer therapy produces undesirable side effects such as cardiotoxicity. To minimize these, attempts have been made to couple the drug with dextran (DEX) and then to encapsulate this drug conjugate in hydrogel nanoparticles. By encapsulation of the drug conjugate in biodegradable, biocompatible long circulating hydrogel nanoparticles, we further improved the therapeutic efficacy of the conjugate. The size of these nanoparticles as determined by quasi-elastic light scattering, was found to be 100+/-10 nm diameter, which favors the enhanced permeability and retention effect (EPR) as observed in most solid tumors. The antitumor effect of these DEX-DXR nanoparticles, was evaluated in J774A.1 macrophage tumor cells implanted in Balb/c mice. The in vivo efficacy of these nanoparticles as antitumor drug carriers, was determined by tumor regression and increased survival time as compared to drug conjugate and free drug. These results suggest that encapsulation of the conjugate in nanoparticles not only reduces the side effects, but also improves its therapeutic efficacy in the treatment of solid tumors. PMID- 11489514 TI - The highly lipophilic DNA topoisomerase I inhibitor DB-67 displays elevated lactone levels in human blood and potent anticancer activity. AB - The novel silatecan 7-t-butyldimethylsilyl-10-hydroxycamptothecin (DB-67) is 25- to 50-times more lipophilic than camptothecin and readily incorporates into lipid bilayers. Using the method of fluorescence anisotropy titration, we determined that DB-67 bound to small unilamellar vesicles composed of dilaurylphosphatidylcholine (DLPC) with an association constant (K value) of 5000 M(-1). This association constant is significantly higher than the K(DLPC) value observed for camptothecin (K(DLPC) value of 110 M(-1)). Using HPLC methods, we demonstrated that the presence of liposomal membranes readily stabilize the lactone form of DB-67. At drug and lipid concentrations of 10 microM and 0.3 mM, respectively, the lactone form of DB-67 persisted in liposome suspension after 3 h of incubation at 37 degrees C. Thus an advantage of a liposomal formulation of DB-67 is that the presence of lipid bilayers assists with stabilizing the key pharmacophore of the agent. The highly lipophilic character of DB-67, in combination with its 10-hydroxy moiety (which functions to enhance lactone stability in the presence of human serum albumin), results in DB-67 having superior stability in human blood with a percent lactone at equilibrium value of 30 [Cancer Res. 59 (1999) 4898; J. Med. Chem. 43 (2000) 3970]. Potent cytotoxicities against a broad range of cancer cells were observed for DB-67, indicating that DB-67 is of comparable potency to camptothecin. The impressive human blood stability and cytotoxicity profiles for DB-67 indicate it is an excellent candidate for comprehensive in vivo pharmacological and efficacy studies. Based on these promising attributes, DB-67 is currently being developed under the NCI RAID program. Due to its potent anti-topoisomerase I activity and its intrinsic blood stability, DB-67 appears as an attractive novel camptothecin for clinical development. PMID- 11489515 TI - Adsorption and desorption of chemotherapeutic drugs from a magnetically targeted carrier (MTC). AB - Magnetically targeted carriers (MTCs) are composite microparticles made from metallic iron and activated carbon. Particles, loaded with doxorubicin in the pharmacy (MTC-DOX), are infused intra-arterially through the artery feeding the tumor. With the aid of an externally positioned permanent dipole magnet, they can be localized and retained within a tumor mass. MTC-DOX is currently in use in a Phase I/II clinical study as a delivery vehicle for doxorubicin in primary hepatocellular carcinoma. The adsorption and desorption of doxorubicin, mitomycin C, camptothecin, methotrexate, verapamil and 9AC onto MTCs have been analyzed. Each of these chemotherapeutic agents has a different mechanism of action, suggesting that some benefit may be derived from combined delivery to a tumor using MTCs and magnetic targeting. Each drug displays different behavior with respect to adsorption and desorption. However, this behavior can be described for each drug with a non-linear thermodynamic model. The thermodynamic model predicts a controlled release rate by adjusting a number of parameters, including initial drug loading concentrations. This is confirmed with in vitro extraction experiments using human plasma as the extraction medium. PMID- 11489516 TI - Molecular vehicle for target-mediated delivery of therapeutics and diagnostics. AB - Selective targeting of therapeutic and diagnostic agents improves their efficacy and minimizes potentially adverse side effects. Existing methods for selective targeting are based on chemical conjugation of therapeutics and diagnostics, or their carriers, to cell-specific targeting molecules (e.g., growth factors, antibodies). These methods are limited by potential damage to targeting molecules that can be inflicted by the conjugation procedure. In addition, conjugation procedures have to be developed on a case-by-case basis. In order to avoid these problems we have developed a new approach to constructing molecular vehicles for target-mediated delivery of therapeutics and diagnostics. In this approach, the targeting molecule is expressed as a fusion protein containing a recognition tag. The recognition tag is defined as a peptide or protein that can bind non covalently another peptide or protein (adapter). In turn, the adapter is chemically conjugated to a carrier of therapeutics or diagnostics. The assembled molecular delivery vehicle contains a carrier-adapter conjugate bound non covalently to a recognition tag fused to the targeting protein. The advantages of this technology are: (i) no chemical modification of targeting molecules, and (ii) universal, 'off-the-shelf' carrier-adapter constructs that can be combined with different fusion targeting proteins. To obtain a proof-of-principle we have constructed VEGF fusion proteins containing a 15-aa S-peptide fragment of RNase A as a recognition tag. Using the S-protein fragment of RNase A as an adapter and polyethylenimine as a DNA carrier we have achieved selective gene delivery to cells overexpressing VEGFR-2. PMID- 11489517 TI - Shiga-like toxin-VEGF fusion proteins are selectively cytotoxic to endothelial cells overexpressing VEGFR-2. AB - Growing endothelial cells at sites of angiogenesis may be more sensitive than quiescent endothelial cells to toxin-VEGF fusion proteins, because they express higher numbers of VEGF receptors. We have constructed, expressed and purified a protein containing the catalytic A-subunit of Shiga-like toxin I fused to VEGF(121) (SLT-VEGF/L). SLT-VEGF/L inhibits protein synthesis in a cell-free translation system and induces VEGFR-2 tyrosine autophosphorylation in cells overexpressing VEGFR-2 indicating that both SLT and VEGF moieties are properly folded in the fusion protein. SLT-VEGF/L selectively inhibits growth of porcine endothelial cells expressing 2-3x10(5) VEGFR-2/cell with an IC(50) of 0.1 nM, and rapidly induces apoptosis at concentrations >1 nM. Similar results are observed with human transformed embryonic kidney cells, 293, engineered to express 2.5x10(6) VEGFR-2/cell. In contrast, SLT-VEGF/L does not affect three different types of endothelial cells (PAE/KDR(low), HUVE, MS1) expressing between 5x10(3) and 5x10(4) VEGFR-2/cell, and quiescent endothelial cells overexpressing VEGFR-2. Growth inhibition and induction of apoptosis by SLT-VEGF/L require intrinsic N glycosidase activity of the SLT moiety, but occur without significant inhibition of protein synthesis. The selective cytotoxicity of SLT-VEGF proteins against growing endothelial cells overexpressing VEGFR-2 suggests that they may be useful in targeting similar cells at sites of angiogenesis. PMID- 11489518 TI - Phage derived peptides for targeting of doxorubicin conjugates to solid tumours. AB - Barriers are frequently hampering targeting of drugs and toxins to solid tumours and their microenvironment. Nano-conjugates are low molecular weight conjugates of a small drug or toxin and a targeting ligand coupled through a cleavable linker group. They offer potential advantages for tumour specific delivery in diffusion-limited situations. We have exploited fd phage-derived peptides for the targeting of low molecular weight drug conjugates to solid tumours. As a model we have chosen doxorubicin conjugates targeted to the transferrin receptor (TfR). A library of phage expressing a cyclic nona-peptide was panned against TfR. The apparent affinity of phages determined by surface plasmon resonance (SPR) increased with each cycle of the panning procedure. After five rounds approximately 80% of phages expressed the same peptide, which mediated a 30-50 fold increased receptor specific cellular uptake of the phages. The corresponding peptide was synthesised using solid phase peptide chemistry on a sulfonamide based safety catch resin. Crude mixtures of the peptide, as well as transferrin itself, were able to inhibit the phage uptake significantly. The doxorubicin conjugate of the peptide containing a cleavable linker was prepared and endosomal uptake confirmed by fluorescence microscopy. PMID- 11489519 TI - Image-guided robotic delivery system for precise placement of therapeutic agents. AB - The effectiveness of conventional solid tumor treatment is limited by the systemic toxicity and lack of specificity of chemotherapeutic agents. Present treatment modalities are frequently insufficient to eliminate competent cancer cells without exceeding the limits of toxicity to normal tissue. The coming generation of cancer therapeutics depends on the precise targeting and sustained release of antitumor agents to overcome these limitations. We are developing an image-guided, robotic system for precise intratumoral placement of anticancer drugs and sustained release devices to advance this new treatment paradigm. The robotic system will use intraoperatively obtained computed tomographic (CT) images from a mobile CT scanner for guidance. The concept is to track patient anatomy and localize instruments using currently available optical tracking technology. Tracking will also be used to register patient anatomy with the images. The physician can then use the registered image to select an appropriate tumor target and entry location and to plan the instrument path. This path will then be transmitted to the robot, which orients and drives the instrument to the desired target under physician control. Achievement of the target is confirmed via intraoperative CT. This system will provide instrument guidance that is precise, direct, and controllable. Error due to poor target visualization and hand unsteadiness should be reduced greatly. The basic components of the system (robot, mobile CT, tracking) have been demonstrated in our laboratory, and the integration of the components is in progress. In future work, we plan to fuse preoperative PET imaging with intraoperative CT to allow functional as well as anatomic image guidance. PMID- 11489522 TI - Modelling aerosol processes related to the atmospheric dispersion of sarin. AB - We have developed mathematical models for evaluating the atmospheric dispersion of selected chemical warfare agents (CWA), including the evaporation and settling of contaminant liquid droplets. The models and numerical results presented may be utilised for designing protection and control measures against the conceivable use of CWA's. The model AERCLOUD (AERosol CLOUD) was extended to treat two nerve agents, sarin and VX, and the mustard agent. This model evaluates the thermodynamical evolution of a five-component aerosol mixture, consisting of two component droplets together with the surrounding three-component gas. We have performed numerical computations with this model on the evaporation and settling of airborne sarin droplets in characteristic dispersal and atmospheric conditions. In particular, we have evaluated the maximum radii (r(M)) of a totally evaporating droplet, in terms of the ambient temperature and contaminant vapour concentration. The radii r(M) range from approximately 15-80 microm for sarin droplets for the selected ambient conditions and initial heights. We have also evaluated deposition fractions in terms of the initial droplet size. PMID- 11489523 TI - Environmental assessment of lubricants before and after wire drawing process. AB - Iron wire drawing processes involve the use of solid lubricants made of powdered raw materials, which lead to industrial wastes after being used. These wastes, based on stearates, have a negative effect on the environment. This study deals with the environmental assessment of some lubricants before and after the wire drawing process in a Spanish factory. The parameters evaluated for this study have been total organic carbon (TOC), mobility of zinc and lead, and ecotoxicity (EC(50)). Results show that wastes have more ecotoxicity than the original lubricants due to the content of metals that lubricants pick up from the wire, as pickling, patenting and galvanising take part in the manufacture. The capture of metallic particles leads to a reduction of TOC and an increase in ecotoxicity. PMID- 11489524 TI - Monitoring of electrokinetic removal of heavy metals in tailing-soils using sequential extraction analysis. AB - This research focused on the monitoring of the electrokinetic removal of heavy metals from tailing-soils, and emphasizes the dependency of removal efficiencies upon their physico-chemical states, as demonstrated by the different extraction methods adopted, which included aqua regia and sequential extraction. The tailing soils examined contained high concentrations of target metal contaminants (Cd=179mgkg(-1), Cu=207mgkg(-1), Pb=5175mgkg(-1), and Zn=7600mgkg(-1)). The removal efficiencies of the different metals were significantly influenced by their speciations, mobilities and affinities (adsorption capacities) in the soil matrix. The removal efficiencies of mobile and weakly bound fractions, such as the exchangeable fraction were more than 90% by electrokinetic treatment, but strongly bound fractions, such as the organically bound species and residual fraction were not significantly removed (less than 30% removal efficiencies). In accordance with the general sequence of mobilities of heavy metals in soils, the removal efficiencies of more mobile heavy metals (Cd, Cu, and Zn) were higher than that of less mobile heavy metal (Pb). PMID- 11489525 TI - Performance evaluation of single-sludge reactor system treating high-strength nitrogen wastewater. AB - In the single-sludge reactor system treating high-strength nitrogen wastewater (similar to anaerobically pretreated piggery wastewater), the NH4(+)-N removal efficiencies (98-82%) are higher than total nitrogen removal efficiencies (71 43%). The mixed liquor recycle ratio only imposes a slight effect on total nitrogen removal efficiency. The alkalinity change data could be used for monitoring and control of the reactor system. To evaluate the performance of the single-sludge reactor system, a simplified nitrification-denitrification model (with nitrification capacity, denitrification capacity, and denitrification potential concepts) and a graphically analytical technique are proposed. It turns out that ammonia nitrification and total nitrogen removal efficiencies are strongly dependent on the process load and reactor configuration, and an optimal operating condition requires a proper match between nitrification and denitrification. PMID- 11489526 TI - Treatability of a simulated disperse dye-bath by ferrous iron coagulation, ozonation, and ferrous iron-catalyzed ozonation. AB - Dyeing and finishing of textile yarns and fabrics are extremely important processes in terms of both quality and environmental concerns. Among the commercial textile dyes, particularly disperse dyestuffs are of environmental interest because of their widespread use, their potential for formation of toxic aromatic amines and their low removal rate during aerobic waste treatment as well as advanced chemical oxidation. Thus, in the present paper ferrous iron coagulation, ozonation and ferrous iron-catalyzed ozonation were employed at varying pH (3-13) and Fe(II)-ion doses (0.09-18mM) for the treatment of a simulated disperse dye-bath (average initial apparent color as absorbance at 566nm=815.4m(-1); COD(0)=3784mgl(-1); TOC(0)=670mgl(-1); BOD(5,0)=58mgl(-1)) that more closely resembled an actual dyehouse effluent than an aqueous disperse dye solution. Coagulation with 5000mgl(-1) FeSO4-7H2O (18mM Fe(2+)) at pH 11 removed up to 97% color and 54% COD, whereas oxidation via ozonation alone (applied ozone dose=2300mgl(-1)) was only effective at pH 3, resulting in 77% color and 11% COD removal. Fe(II)-ion-catalyzed ozonation (3.6mM Fe(2+) at pH 3; Fe(2+):O3 molar ratio 1:14) eliminated 95% color and 48% COD and appeared to be the most attractive option among the investigated chemical treatment methods as for its applicability at the natural acidic pH of the disperse dye-bath effluent and at relatively low Fe(2+)-ion doses as compared to ferrous sulfate coagulation. However, no TOC reduction was observable for ozonation and catalytic ozonation at the investigated reaction conditions (14gl(-1) O3 at pH 3). An average six-fold enhancement in the biodegradability parameter of the synthetic dye wastewater expressed in terms of the BOD(5)/COD ratio could be achieved by the investigated chemical treatment methods. PMID- 11489527 TI - Risk-based monitored natural attenuation--a case study. AB - The term "monitored natural attenuation" (MNA) refers to a reliance on natural attenuation (NA) processes for remediation through the careful monitoring of the behavior of a contaminant source in time and space domains. In recent years, policymakers are shifting to a risk-based approach where site characteristics are measured against the potential risk to human health and the environment, and site management strategies are prioritized to be commensurate with that risk. Risk based corrective action (RBCA), a concept developed by the American Society for Testing Materials (ASTM), was the first indication of how this approach could be used in the development of remediation strategies. This paper, which links ASTM's RBCA approach with MNA, develops a systematic working methodology for a risk based site evaluation and remediation through NA. The methodology is comprised of seven steps, with the first five steps intended to evaluate site characteristics and the feasibility of NA. If NA is effective, then the last two steps will guide the development of a long-term monitoring plan and approval for a site closure. This methodology is used to evaluate a site contaminated with oil from a pipeline spill. The case study concluded that the site has the requisite characteristics for NA, but it would take more than 80 years for attenuation of xylene and ethylbenzene, as these chemicals appear in the pure phase. If fast remediation is sought, then efforts should be made to remove the contaminant from the soil. Initially, the site posed a serious risk to both on-site and off-site receptors, but it becomes acceptable after 20 years, as the plume is diluted and drifts from its source of origin. PMID- 11489528 TI - Estimation of the atmospheric corrosion on metal containers in industrial waste disposal. AB - Solid industrial waste are often stored in metal containers filled with concrete, and placed in well-aerated warehouses. Depending on meteorological conditions, atmospheric corrosion can induce severe material damages to the metal casing, and this damage has to be predicted to achieve safe storage. This work provides a first estimation of the corrosivity of the local atmosphere adjacent to the walls of the container through a realistic modeling of heat transfer phenomena which was developed for this purpose. Subsequent simulations of condensation/evaporation of the water vapor in the atmosphere were carried out. Atmospheric corrosion rates and material losses are easily deduced. For handling realistic data and comparison, two different meteorological contexts were chosen: (1) an oceanic and damp atmosphere and (2) a drier storage location. Some conclusions were also made for the storage configuration in order to reduce the extent of corrosion phenomena. PMID- 11489529 TI - Removal of Cu, Pb and Zn in an applied electric field in calcareous and non calcareous soils. AB - The pH dependency of the removal of Cu, Zn and Pb by electrodialytic soil remediation from different industrially polluted soils was examined. From 18 experiments performed with five different soils, it was found that the order of mobilization due to a pH decrease was Zn>Cu>Pb. It was found, too, that each of the elements was removed at higher soil pH in calcareous soils (about 12% carbonates) than in soils with a carbonate content of less than 3.7%. In soils rich in carbonates, precipitation of heavy metal carbonates is an important retention mechanism and the heavy metal carbonates are dissolved at higher pH values than the pH at which heavy metals are desorbed in non-calcareous soils. Thus, the relation between the soil pH and the mobility of the heavy metal in the electric field is not only dependent on the heavy metal in focus, but also on the fraction of the heavy metal precipitated as carbonates. PMID- 11489530 TI - Production of granular activated carbon from waste walnut shell and its adsorption characteristics for Cu(2+) ion. AB - Production of granular activated carbon by chemical activation has been attempted employing walnut shells as the raw material. The thermal characteristics of walnut shell were investigated by TG/DTA and the adsorption capacity of the produced activated carbon was evaluated using the titration method. As the activation temperature increased, the iodine value increased. However, a temperature higher than 400 degrees C resulted in a thermal degradation, which was substantiated by scanning electron microscopy (SEM) analysis, and the adsorption capacity decreased. Activation longer than 1h at 375 degrees C resulted in the destruction of the microporous structure of activated carbon. The iodine value increased with the increase in the concentration of ZnCl2 solution. However, excessive ZnCl2 in the solution decreased the iodine value. The extent of activation by ZnCl2 was compared with that by CaCl2 activation. Enhanced activation was achieved when walnut shell was activated by ZnCl2. Applicability of the activated carbon as adsorbent was examined for synthetic copper wastewater. Adsorption of copper ion followed the Freundlich model. Thermodynamic aspects of adsorption have been discussed based on experimental results. The adsorption capacity of the produced activated carbon met the conditions for commercialization and was found to be superior to that made from coconut shell. PMID- 11489531 TI - Remediation of TCE contaminated soils by in situ EK-Fenton process. AB - The treatment performance and cost analysis of in situ electrokinetic (EK)-Fenton process for oxidation of trichloroethylene (TCE) in soils were evaluated in this work. In all experiments, an electric gradient of 1V/cm, de-ionized water as the cathode reservoir fluid and a treatment time of 10 days were employed. Treatment efficiencies of TCE were evaluated in terms of the electrode material, soil type, catalyst type, and catalyst dosage and granular size if applicable. Test results show that graphite electrodes are superior to stainless steel electrodes. It was found that the soil with a higher content of organic matter would result in a lower treatment efficiency (e.g. a sandy loam is less efficient than a loamy sand). Experimental results show that the type of catalyst and its dosage would markedly affect the reaction mechanisms (i.e. "destruction" and "removal") and the treatment efficiency. Aside from FeSO4, scrap iron powder (SIP) in the form of a permeable reactive wall was also found to be an effective catalyst for Fenton reaction to oxidize TCE. In general, the smaller the granular size of SIP, the lower the overall treatment efficiency and the greater the destruction efficiency. When a greater quantity of SIP was used, a decrease of the overall treatment efficiency and an increase of percent destruction of TCE were found. Experimental results have shown that the quantity of electro-osmotic (EO) flow decreased as the quantity of SIP increased. It has been verified that the treatment performances are closely related to the corresponding EO permeability. Results of the cost analysis have indicated that the EK-Fenton process employed in this work is very cost-effective with respect to TCE destruction. PMID- 11489532 TI - Beta3, a novel auxiliary subunit for the voltage gated sodium channel is upregulated in sensory neurones following streptozocin induced diabetic neuropathy in rat. AB - In the present study we have used in situ hybridization to examine the changes in mRNA expression of the voltage gated sodium channel subunits beta1 and beta3, which occur in response to streptozocin induced diabetic neuropathy. Under control conditions beta1 mRNA was detected throughout the spinal cord and in large dorsal root ganglion (DRG) Abeta fibres whilst beta3 mRNA was expressed exclusively in the layers I/II and X of the spinal cord and in small DRG c fibres. Following streptozocin treatment, the expression of beta1 mRNA remained unchanged in both the spinal cord and DRG whilst beta3 message was significantly increased in both the spinal cord and in medium diameter Adelta type DRG neurones. In conclusion, the present study illustrates that the development of the neuropathic pain state is associated with distinct changes in the pattern of beta3 subunit expression and that these changes appear to be specific to the neuropathic pain state induced. PMID- 11489533 TI - Nitric oxide modulates spontaneous cord dorsum potentials in the cat spinal cord. AB - A previous study has shown that lumbar spontaneous cord dorsum potentials (CDPs) are produced by background activity of a neuronal ensemble located in the dorsal horn. Here, the effects produced by intravenous application of the nitric oxide synthase inhibitor L-N(G)-nitro arginine (L-NOARG, 100 microg/kg) and of the nitric oxide donor 3-morpholinosydnonimine hydrochloride (SIN-1, 500 microg/kg) on spontaneous CDPs were examined. Experiments were performed on pentobarbitally anesthetized, paralyzed and spinalized cats. The amplitude of spontaneous CDPs increased after L-NOARG, however, decreased after SIN-1. These observations suggest that electrical activity of dorsal horn neurones generating spontaneous CDPs is dependent on nitric oxide production. PMID- 11489534 TI - Inhibition of neuronal nitric oxide synthase enhances cell proliferation in the dentate gyrus of the adrenalectomized rat. AB - Recent studies have demonstrated that the elimination of adrenal steroids by an adrenalectomy (ADX) increases the expression of neuronal nitric oxide synthase (NOS), and that it increases cell proliferation in the rat dentate gyrus. However, no evidence has been presented to date which indicates that NO regulates cell proliferation in the dentate gyrus of the adult rats. In this study, the effect of blocking NO production on ADX-induced increase of cell proliferation and serotonergic innervation was examined in the rat dentate gyrus. 7 nitroindazole (7-NI; 30 mg/kg, intraperitoneally), a selective inhibitor of neuronal NOS, was injected 1 day before an ADX and then once every 24 h for 4 days after the ADX subsequently. The proliferating cells were identified with 5 bromo-2-deoxyuridine (BrdU) immunostaining. Long-term inhibition of the neuronal NOS by 7-NI markedly increased the BrdU-labeled cell population density 4-18-fold in the dentate gyrus of the adrenalectomized rats compared to that in the vehicle injected adrenalectomized rats. Immunoreactivity of serotonin, known as a mediator of granule cell genesis, was detected only in the dentate gyrus of 7-NI injected adrenalectomized rats. These results indicate that NO may be involved in the cell proliferation in the dentate gyrus of the adrenalecomized rat and that serotonin may mediate the regulatory effect of NO on the cell proliferation in rat dentate gyrus. PMID- 11489535 TI - Interleukin-6 protects rat PC12 cells from serum deprivation or chemotherapeutic agents through the phosphatidylinositol 3-kinase and STAT3 pathways. AB - The mechanisms underlying the anti-apoptotic action of interleukin (IL)-6 on hematopoietic cells have been extensively studied, but those in the case of neuronal cells have been poorly reported. We investigated the effect of IL-6 on the survival of rat PC12 pheochromocytoma cells and analyzed the signaling pathways of the cytokine by means of some kinase inhibitors. IL-6 protects PC12 cells from the death induced by serum deprivation or anticancer agents, such as cisplatin, paclitaxel and 5-fluorouracil. Phosphatidylinositol (PI)3-kinase inhibitors (LY294002 and wortmannin) but not a mitogen-activated protein kinase kinase inhibitor (PD98059) completely suppressed the IL-6-promoted survival of the cells. A Janus tyrosine kinase 2 inhibitor (tyrphostin AG490) suppressed the phosphorylation of signal transducers and activators of transcription (STAT)3 and only partially inhibited the anti-apoptotic activity of IL-6. IL-6 stimulated phosphorylation of Akt, a downstream effector of PI3 kinase, and in the presence of LY294002, the phosphorylation of Akt was reduced to basal level. These results suggest that the signaling pathway for the anti-apoptotic effect of IL-6 in PC12 cells is mediated in major part by activation of the PI3-kinase/Akt pathway and thus is different from that in hematopoietic cells. PMID- 11489536 TI - A comparison of rheumatoid arthritis and fibromyalgia patients and healthy controls exposed to a pulsed (200 microT) magnetic field: effects on normal standing balance. AB - Specific weak time varying pulsed magnetic fields (MF) have been shown to alter animal and human behaviors, including pain perception and postural sway. Here we demonstrate an objective assessment of exposure to pulsed MF's on Rheumatoid Arthritis (RA) and Fibromyalgia (FM) patients and healthy controls using standing balance. 15 RA and 15 FM patients were recruited from a university hospital outpatient Rheumatology Clinic and 15 healthy controls from university students and personnel. Each subject stood on the center of a 3-D forceplate to record postural sway within three square orthogonal coil pairs (2 m, 1.75 m, 1.5 m) which generated a spatially uniform MF centered at head level. Four 2-min exposure conditions (eyes open/eyes closed, sham/MF) were applied in a random order. With eyes open and during sham exposure, FM patients and controls appeared to have similar standing balance, with RA patients worse. With eyes closed, postural sway worsened for all three groups, but more for RA and FM patients than controls. The Romberg Quotient (eyes closed/eyes open) was highest among FM patients. Mixed design analysis of variance on the center of pressure (COP) movements showed a significant interaction of eyes open/closed and sham/MF conditions [F=8.78(1,42), P<0.006]. Romberg Quotients of COP movements improved significantly with MF exposure [F=9.5(1,42), P<0.005] and COP path length showed an interaction approaching significance with clinical diagnosis [F=3.2(1,28), P<0.09]. Therefore RA and FM patients, and healthy controls, have significantly different postural sway in response to a specific pulsed MF. PMID- 11489537 TI - Anticholinesterase treatment of chicken retinal cells increases acetylcholinesterase protein independently of protein kinase C. AB - It has been reported that anticholinesterase exposure, e.g. by environmental toxins or nerve gases, can increase acetylcholinesterase (AChE) protein, possibly as an autoregulatory stress response. We earlier have transfected retinal cells of the chick embryo with a pSVK3-AChE(rab)-cDNA vector to heterologously express rabbit AChE, which concomitantly also increased AChE protein from chick. To analyse further the cell-internal pathways of these different paradigms (anticholinesterase treatment vs. AChE transfection) which both lead to an AChE increase, we here show that AChE overexpression by transfection leads to an increase in protein kinase C (PKC). Most remarkably, when cells independently of, or in addition to their transfection are treated with 10 microM of the AChE inhibitor BW284c51, AChE protein levels are much more dramatically increased up to 20-fold. This treatment, however, does not affect PKC. These data show that (i) retinal cells respond to anticholinesterase insult by a massive increase of AChE protein; (ii) the response to BW284c51 is not PKC-mediated; and (iii) both strategies of AChE increase follow different cell-internal pathways, their effects being additive. The ecological and biomedical implications of these findings are briefly discussed. PMID- 11489538 TI - Changes in the levels of nitric oxide synthase and protein kinase C gamma following kainic acid receptor activation in the rat spinal cord. AB - In this study, we evaluated the levels of nitric oxide synthase, both neuronal and induced (nNOS and iNOS, respectively), cyclooxygenase-1 and 2 (COX-1 and COX 2) and protein kinase C gamma (PKCgamma) and correlated these with algogenic behavior following spinal kainic acid (KA) receptor activation in rats. Thirty adult male Sprague-Dawley rats were randomly assigned into six groups (n=5). Groups A, B, and C received 0.5 g kainic acid intrathecally and were analyzed at 3, 6, 24 h after injection, respectively. Groups D, E, and F received saline and were analyzed at 3, 6, 24 h after injection, respectively. We observed for behavioral changes in the rats following intrathecal KA injection and analyzed the protein levels of NOS, COX and PKCgamma by Western blotting techniques. Importantly, we clarified the potential roles of PKCgamma in the regulation of nNOS and COX-2 following intrathecal injection with KA in the rat spinal cord. COX-2 protein was detected but not significantly changed in the lumbosacral spinal cord at 3, 6, and 24 h following intrathecal KA injection (P>0.05). In contrast, nNOS protein was detected at higher levels in comparison with normal spinal cord at 6 and 24 h after intrathecal administration of KA (P<0.05). PKCgamma also increased significantly at 3, 6, and 24 h after intrathecal KA injection when compared with the baseline level (P<0.05). On the other hand, COX 1 and iNOS were not detected in either normal or KA treated spinal cords. These results provide strong in vivo evidence to support the idea that nNOS but not COX 2, plays an important role in spinal KA receptor activation. Furthermore, up regulation of PKCgamma is involved in KA induced algogenic behavior in rats. PMID- 11489539 TI - Cerebello-thalamo-cortical projections to the posterior parietal cortex in the macaque monkey. AB - The cerebello-thalamo-posterior parietal cortical projections were investigated electrophysiologically and morphologically in macaque monkeys. In anesthetized monkeys, electrical stimulation of every cerebellar nucleus evoked marked surface positive, depth-negative (s-P, d-N) cortical field potentials in the superior parietal lobule and the cortical bank of the intraparietal sulcus, but no responses in the inferior parietal lobule. Tract-tracing experiments combining the anterograde method with the retrograde one indicated that the interposed and lateral cerebellar nuclei projected to the posterior parietal cortex mainly through the nucleus ventral lateralis caudalis of the thalamus. The significance of the projections is discussed in connection with cognitive functions. PMID- 11489540 TI - Distribution and characterization of vanilloid receptors in the rat stomach. AB - The cloned vanilloid receptor-1 (VR1) is recognized as a common molecular target for protons, noxious heat, and vanilloids. The presence of VR1 in the dorsal root, trigeminal, and nodose ganglia has been firmly established, but it is unclear in the gut, despite this VR1 may be important for gastric mucosal homeostasis. In this study we used an antibody and a radioligand to show the distribution of vanilloid receptors (VRs) in rat stomach and to characterize it. The deafferentiation of capsaicin-sensitive nerves in rats was induced by consecutive injections of capsaicin. VR1-immunopositive nerve endings were predominantly found in the mucous neck cells of the proliferation zone, and around blood vessels in the submucosa. Radioreceptor assay using [3H] resiniferatoxin (RTX) revealed the existence of high affinity and single-class binding site in the membrane fractions of the mucosa. Capsaicin completely inhibited the specific binding of [3H]-RTX. Both the VR1 immunoreactivity and the receptor density of [3H]-RTX binding sites significantly reduced by the application of capsaicin for prolonged periods of time in the mucosa of rats. Our results indicate that VRs are expressed in the rat stomach, and suggest that they may be involved in mucosal protection by increasing cell proliferation and blood flow. PMID- 11489541 TI - Effects of estrogen and progesterone on the expression of connexin-36 mRNA in the suprachiasmatic nucleus of female rats. AB - To determine the effects of ovarian steroid hormones on gap junction communication in the suprachiasmatic nucleus (SCN), we examined the effects of estrogen and/or progesterone on the expression of connexin-36 mRNA in the SCN and cerebral cortex (CX) of female rats. Ovariectomized adult rats were injected with 20 microg 17beta-Estradiol or sesame oil 48 h before sacrifice and further injected with 1.5 mg progesterone or sesame oil 24 h before sacrifice. Northern blot revealed that estrogen significantly increased the expression of connexin-36 mRNA in the SCN and this increase was inhibited by progesterone. On the other hand, the connexin-36 mRNA level in the CX was not affected by estrogen or progesterone. These results suggest that the gap junction with connexin-36 in the SCN is specifically regulated by ovarian steroid hormones of female rats. PMID- 11489542 TI - Asymmetry of parietal lobe activation during piano performance: a high field functional magnetic resonance imaging study. AB - Functional asymmetry of the parietal lobes during piano performance was assessed utilizing independent component-cross correlation-sequential epoch analysis of functional magnetic resonance imaging time series. Eight right handed musically trained subjects played the piano with their right hand, left hand, or both hands as cued by visually presented musical scores. The areas activated included the posterior parietal cortex (PPC) and the primary sensorimotor areas (SM1). While unilateral SM1 activation was correlated to motion of the corresponding contralateral hand, PPC activation was correlated to piano performance irrespective of hand modality. Furthermore, PPC activation exhibited significant asymmetry, with left hemisphere dominance. The results indicate that the left parietal lobe plays a significant role in the cortical processes of piano performance. PMID- 11489543 TI - Memantine, but not dizocilpine, ameliorates cognitive deficits in adult rats withdrawn from chronic ingestion of alcohol. AB - Adult rats were given a 20% ethanol solution as their only source of fluid for 6 months and then withdrawn from alcohol. During the first 4 weeks of the withdrawal period, animals were intraperitoneally injected with either memantine (20 mg/kg bolus followed by 1 mg/kg every 12 h) or dizocilpine (MK-801; 0.1 mg/kg every 12 h), both of which are antagonists of N-methyl-D-aspartate receptors. Ten weeks after initiation of the withdrawal procedure, cognitive status of animals was assessed using the Morris water maze. Withdrawal from alcohol produced robust deficits in the performance of rats on the acquisition task and on the probe trial. Treatment with memantine resulted in a complete reversal of these behavioral impairments. In contrast, treatment with MK-801 was found to be ineffective in preventing cognitive alterations associated with chronic alcohol consumption and withdrawal. PMID- 11489544 TI - Fructose- 1,6-bisphosphate did not affect hippocampal neuronal damage caused by 10 min of complete umbilical cord occlusion in fetal sheep. AB - Fructose-1, 6-bisphosphate (FBP) has a neuroprotective effect in neonatal and adult rats. The purpose of this study was to examine the effects of FBP on hippocampal neuronal damage in fetal sheep asphyxiated by 10 min of complete umbilical cord occlusion. Thirteen fetal sheep at 124 days of gestation were surgically instrumented with catheters. Cardiorespiratory parameters were monitored, and biochemical analyses were performed with the blood samples. During the insult seven fetuses were given FBP (500 mg/kg) and six were given iso osmotic saline, and hippocampal neuronal damage was examined histologically and scored. Cardiorespiratory changes were the same in both groups, and there was no neuroprotective effect of FBP in this study. However the decrease of serum total Ca level implied the Ca- chelating effect of FBP. PMID- 11489545 TI - Spatial and temporal distribution of pituitary adenylate cyclase activating polypeptide in gerbil global cerebral ischemia. AB - In the present study, we used immunohistochemistry to investigate the effects of transient global ischemia on pituitary adenylate cyclase activating polypeptide (PACAP) immunoreactivity in gerbil brain regions. PACAP immunoreactivity had significantly decreased in the pyramidal cells of CA1 subfield susceptible to ischemic insult at 1-4 days after transient global ischemia. On the contrary, PACAP immunoreactivity had not changed in the pyramidal cell bodies of more resistant CA3 subfield after ischemic injury. In the Purkinje cell layer, PACAP immunoreactivity had significantly decreased 1 day following transient ischemia, and had increased 2 days after ischemia. The first demonstration of the postischemic localizations of PACAP should allow us to gain a more fundamental rationale for developing methods of treating ischemic brain damage with neuroprotective peptides such as PACAP. PMID- 11489546 TI - Nitric oxide produces different actions in different areas of the periaqueductal grey in cats. AB - In 20 urethane-anaesthetised cats, microinjection of N-methyl-D-aspartate (NMDA) into the intermediate-lateral (IL-) or the dorsolateral (DL-) periaqueductal grey areas (PAG) of the midbrain elicited similar patterns of cardiovascular responses: increases in mean systemic arterial pressure (MSAP), heart rate (HR) and mean blood flows (F) of the common carotid and femoral arteries, accompanied with a 'hissing-howling' response. Similar increases in MSAP and Fs were induced by microinjections of S-nitroso-N-acetylpenicillamine (SNAP), a potent nitric oxide (NO) donor, on the IL-PAG (A2.5-A0.5). In contrast, microinjections of N(G) nitro-L-arginine methyl ester (L-NAME), a NO synthase inhibitor, in the same area elicited a decrease in MSAP and Fs. On the other hand, microinjections of either SNAP or L-NAME reversed the original IL-PAG responses when injected into the DL PAG. Pretreatments with SNAP significantly inhibited NMDA-induced responses in the DL-PAG but potentiated such responses in the IL-PAG. In contrast, pretreatments with L-NAME potentiated the NMDA-induced responses in the DL-PAG but inhibited such responses in the IL-PAG. These data suggest that NO may be a neurotransmitter or neuromodulator that exerts differential function in different defense areas, namely the IL- and the DL-PAG. PMID- 11489547 TI - Increase of the ornithine decarboxylase/polyamine system and transglutaminase upregulation in the spinal cord of aged rats. AB - We have investigated changes in ornithine decarboxylase (ODC) activity and in polyamine levels in the central nervous system of aged rats. We measured a significant increase of ODC catalytic activity in the spinal cord from 30 month old rats (+105%) as compared to 4 month-old rats. No changes were noticed in the cerebellum, cortex and hippocampus from the same animals. A related putrescine increase was measured in the spinal cord of 30 month-old rats (+168%), together with a smaller increase of spermidine (+33%). A parallel increase (+78%) of the Ca2+-dependent transglutaminase activity was detected in the spinal cord of 30 month-old rats, while no changes were apparent in the cortex and cerebellum. Our observations indicate a possible role of the ODC/polyamine system during the normal process of ageing in rats and point to the spinal cord as the most sensitive area for this kind of modification. A possible role of protein polyamination by transglutaminase is discussed. PMID- 11489548 TI - The plant flavonoid wogonin suppresses death of activated C6 rat glial cells by inhibiting nitric oxide production. AB - Flavonoids are a group of low molecular weight polyphenolic compounds derived from plants. 5,7-dihydroxy-8-methoxyflavone (Wogonin), a flavonoid originated from the root of Scutellaria baicalensis Georgi, has been shown to exert various anti-inflammatory effects such as inhibition of nitric oxide (NO) and prostaglandin E2 production in macrophages. Because glial cells have been previously shown to undergo NO-dependent apoptosis upon inflammatory activation and this auto-regulatory process may be negatively affected by exogenous factors possessing anti-inflammatory activities, we examined the effects of wogonin on NO production and activation-induced cell death of C6 rat glial cells. Activation of C6 glial cells with lipopolysaccharide (LPS), interferon-gamma, and tumor necrosis factor-alpha induced NO production followed by cell death. Pretreatment of C6 cells with wogonin before LPS and cytokine treatment dose-dependently inhibited NO production as well as death of activated C6 cells. Wogonin-mediated inhibition of NO production was accompanied by suppression of inducible nitric oxide synthase (iNOS) protein induction and nuclear factor kappa B (NF-kappaB) reporter activity. Wogonin, however, did not affect a NO donor-induced cytotoxicity. Taken together, our results indicate that wogonin inhibits activation-induced death of C6 glial cells by suppressing NO production, and these inhibitory effects of wogonin on NO production are exerted through inhibition of NF-kappaB-mediated iNOS induction. PMID- 11489549 TI - Inhibition of RNA synthesis differentially affects in vitro melatonin release from the pineal organs of ayu (Plecoglossus altivelis) and rainbow trout (Oncorhynchus mykiss). AB - The effects of actinomycin D (RNA synthesis inhibitor) and cycloheximide (protein synthesis inhibitor) on melatonin release from the cultured pineal organ of two teleosts with or without the circadian regulation of melatonin production (ayu Plecoglossus altivelis and rainbow trout Oncorynchus mykiss, respectively) were investigated. Actinomycin D decreased melatonin release from the pineal organ during the dark phase but there was a significant difference between the two species (22.2% for ayu and 59.1% for trout as compared with the respective control). This difference might be due to whether the circadian regulation via gene transcription of melatonin synthesis exists or not. On the other hand, cycloheximide decreased melatonin release to approximately 1% in both species, indicating that the fish pineal organ requires de novo protein synthesis to maintain rhythmic melatonin release. PMID- 11489550 TI - Formulation and stability testing of photolabile drugs. AB - Exposure of a drug to irradiation can influence the stability of the formulation, leading to changes in the physicochemical properties of the product. The influence of excipients of frequently used stabilizers is often difficult to predict and, therefore, stability testing of the final preparation is important. The selection of a protective packaging must be based on knowledge about the wavelength causing the instability. Details on drug photoreactivity will also be helpful in order to minimize side-effects and/or optimize drug targeting by developing photoresponsive drug delivery systems. This review focuses on practical problems related to formulation and stability testing of photolabile drugs. PMID- 11489551 TI - Cyclodextrins in topical drug formulations: theory and practice. AB - Cyclodextrins are cyclic oligosaccharides with a hydrophilic outer surface and a somewhat lipophilic central cavity. Cyclodextrins are able to form water-soluble inclusion complexes with many lipophilic water-insoluble drugs. In aqueous solutions drug molecules located in the central cavity are in a dynamic equilibrium with free drug molecules. Furthermore, lipophilic molecules in the aqueous complexation media will compete with each other for a space in the cavity. Due to their size and hydrophilicity only insignificant amounts of cyclodextrins and drug/cyclodextrin complexes are able to penetrate into lipophilic biological barriers, such as intact skin. In general, cyclodextrins enhance topical drug delivery by increasing the drug availability at the barrier surface. At the surface the drug molecules partition from the cyclodextrin cavity into the lipophilic barrier. Thus, drug delivery from aqueous cyclodextrin solutions is both diffusion controlled and membrane controlled. It appears that cyclodextrins can only enhance topical drug delivery in the presence of water. PMID- 11489552 TI - Polymethyacrylate based microparticulates of insulin for oral delivery: preparation and in vitro dissolution stability in the presence of enzyme inhibitors. AB - The purpose of this investigation was to (a) evaluate the coprecipitation technique for preparing microparticulates of insulin, (b) study the effect of variables such as addition of salts in the precipitating medium and ratio of polymeric solution to volume of precipitating medium on the dissolution and encapsulation efficiency of insulin microparticulates, and (c) evaluate the in vitro enzymatic dissolution stability of insulin microparticulates in the presence of chicken ovomucoid (CkOVM) and duck ovomucoid (DkOVM) as inhibitors. Insulin dissolved in 0.01 N HCl was mixed with alcohol USP to get a final concentration of 32% v/v. Eudragit L100, a representative polymethyacrylate polymer, was then dissolved in this solution which was transferred to a beaker containing cold water with homogenization to obtain microparticulates. Dissolution studies were carried out in pH 6.8 phosphate buffer using a 100-ml conversion kit in a standard dissolution assembly. Dissolution stability of microparticulates was evaluated in the presence of 0.5 microM trypsin and 0.l microM chymotrypsin at various ratios of CkOVM and DkOVM. The results indicated that insulin microparticulates could be prepared using the coprecipitation technique with high encapsulation efficiency by proper selection of experimental conditions and amount of polymer. Presence of salts in the precipitating medium decreased the dissolution of insulin from the microparticulates. As the ratio of precipitating medium with respect to the polymeric solution was increased, the encapsulation efficiency increased. In dissolution stability experiments, insulin was not detected in the presence of enzymes alone. When CkOVM and DkOVM were incorporated, the stability of insulin increased significantly in a concentration dependent fashion. PMID- 11489553 TI - Solubilization of NSC-639829. AB - Solubilization using pH combined with cosolvents, surfactants, and complexants are investigated for NSC-639829, an investigational anti-tumor agent. The intrinsic solubility of the drug is approximately 30 ng/ml and it has an ionizable dimethyl aniline group with an approximate base pK(a) of 5. Samples buffered at pH 1.0, 2.0, and 7.0 with various concentrations of the solubilizing agents were used to study the solubilization of NSC-629829 when present as charged and uncharged species. The solubilization of NSC-639829 was found to be much more effective when the drug was present primarily in ionized form. At pH values 1.0 and 2.0 where the surfactant (SLS) and complexant (SBEbetaCD) carried a negative charge enhanced solubilities of more than a million-fold were observed for the drug. PMID- 11489554 TI - In vitro and in vivo evaluation of carbamazepine-PEG 6000 solid dispersions. AB - The present work extended previous physico-chemical investigations on the effects of solid dispersion on the solubility, the dissolution rate and the pharmacokinetic profile of carbamazepine. Solubility studies showed a linear increase in carbamazepine solubility with the increase of PEG 6000 concentration. There is no marked difference between physical mixtures and solid dispersions for the enhancement of carbamazepine solubility by PEG 6000. Less than 60% of pure carbamazepine was dissolved in 90 min. Physical mixtures (carbamazepine phase III) and solid dispersions (carbamazepine phase II) dissolution rates were higher in comparison of the parent drug. The dissolution of carbamazepine phase III was more pronounced than that evoked by the phase II. The dissolution profiles indicated that the percentage of the drug dissolved was dependent on the proportion of PEG 6000. In solid dispersions there was a remarkable enhancement in the dissolution rates of the drug in the vicinity of the eutectic composition as compared with those of corresponding physical mixtures. Hence, the optimum value for the solid dispersion was 80.5+/-1.7% of carbamazepine having dissolved within the first 10 min compared to 40+/-1% for the corresponding physical mixtures of the same composition. Statistical analysis of pharmacokinetic parameters confirmed that the carbamazepine:PEG 6000 binary systems displayed higher bioavailability of the drug than the pure carbamazepine. The area under the curve (AUC) values highlighted the evidence that only slight differences in the bioavailability of the drug occur between physical mixtures and solid dispersions prepared at the 80:20 and 50:50 drug:carrier compositions. However, the mean normalized plasma concentrations showed that standard error deviations are rather wide intervals for pure drug and physical mixtures in comparison to solid dispersions. One additional interesting point to consider is the disappearance of the multiple peaks on the individual kinetic curves of the 50:50 solid dispersion composition. Furthermore, our investigations have highlighted the interest of solid dispersions prepared at <>-eutectic composition as our preliminary data show that the plasma concentration (C(5h)) of the drug for the 15:85 dispersed sample containing 150 mg of carbamazepine is not significantly different from that obtained for the 50:50 dispersed sample containing 300 mg of the drug. PMID- 11489555 TI - Dissolution properties and anticonvulsant activity of phenytoin-polyethylene glycol 6000 and -polyvinylpyrrolidone K-30 solid dispersions. AB - Solid dispersions of phenytoin in polyethylene glycol 6000 and polyvinylpyrrolidone K-30 with different drug-to-carrier ratios were prepared by the solvent method with the aim of increasing dissolution rate and bioavailability of the drug. These new formulations were characterized in the solid state by FT-IR spectroscopy, X-ray powder diffraction, and differential scanning calorimetry. Drug solubility and dissolution rate are improved by these formulations, particularly with SDPEG 1/20 and SDPVP 1/20 systems. Storage was found to influence the stability of the solid dispersions. By maximal electroshock test, it was found that the intraperitoneal administration in mice of the SDPEG 1/20 and SDPVP 1/20 systems exhibited anticonvulsant activity similar to diphenylhydantoin sodium salt. PMID- 11489556 TI - Lipid extraction and transport of hydrophilic solutes through porcine epidermis. AB - The purpose of this study was to investigate the effect of delipidization of the stratum corneum (SC) on the in vitro percutaneous absorption of hydrophilic solutes (i.e. water, urea, and inulin). Fourier transform infrared (FT-IR) spectroscopy was employed to study the extent of delipidization of porcine SC due to chloroform:methanol (2:1) (C:M (2:1)) treatments for various time periods. In vitro percutaneous absorption of [3H] water, [14C] urea, and [3H] inulin were studied through C:M (2:1) treated epidermis in Franz diffusion cells. There was a greater decrease in peak areas of the asymmetric and symmetric C-H stretching absorbances (i.e. increase in lipid extraction) with increasing exposure times of the SC with C:M (2:1). After 40-min treatment, asymmetric and symmetric C-H stretching peak area showed a decrease of 75.9 and 89.9%, respectively. The permeability coefficient of water, urea, and inulin increased with increasing lipid extraction. Enhancement in the permeability coefficient, through 40 min C:M (2:1) treated epidermis in comparison to the control, for water, urea, and inulin was 48.72, 215.65, and 3.90, respectively. Log (permeability coefficient) and log (mol. wt.) for test solutes and leuprolide acetate were found to be inversely related (R(2)=0.9974). In conclusion, this study implies that penetration enhancers that are safe and extract the SC lipids can be selected in order to enhance the percutaneous absorption of polar solutes through the skin. PMID- 11489557 TI - Influence of various drugs on the glass transition temperature of poly(vinylpyrrolidone): a thermodynamic and spectroscopic investigation. AB - PURPOSE: To determine the influence of hydrogen bonding and solubility parameter on the glass transition temperature (T(g)) of various drug-poly(vinylpyrrolidone) blends. METHODS: The T(g) of PVP films containing either acetaminophen, naproxen, salicylamide, carbamazepine, griseofulvin or propranolol hydrochloride were measured using differential scanning calorimetry. Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction was used to characterize the specific interactions between the drug-PVP blends and the physical state of the films, respectively. The total solubility parameter and its individual components were calculated using the method of Van Krevelen. RESULTS: Salicylamide displayed the greatest plasticizing effect, depressing the T(g) to the minimum. This was consistent with the FTIR data, which indicated the presence of hydrogen bonding with PVP. Griseofulvin showed the least plasticizing effect due to lack of interaction with PVP. All the drugs except griseofulvin were amorphous within the film up to 30% (w/w) drug composition. The correlation between the various components of the solubility parameters and the plasticizing effect of drugs was very poor. CONCLUSIONS: Spectroscopic investigation for the presence of interaction between the drugs and PVP proved to be extremely predictive of the plasticizing effect of various drugs. In contrast, solubility parameters appeared to be far less sensitive indicators of drug-PVP miscibility. PMID- 11489558 TI - Cationized human serum albumin as a non-viral vector system for gene delivery? Characterization of complex formation with plasmid DNA and transfection efficiency. AB - Cationized human serum albumin (cHSA) could serve as a potential non-viral vector system for gene delivery. Native human serum albumin was cationized by covalent coupling of hexamethylenediamine to the carboxyl groups resulting in a shift of the isoelectric point from pH 4-5 to 7-9. The cationized albumin underwent spontaneous self-assembly with DNA as demonstrated by retardation of CMV-nlacZ plasmid in agarose gel electrophoresis. Photon correlation spectroscopy showed a decrease of complex size with increasing cHSA/plasmid ratios. Under optimized conditions complexes were formed with 230-260 nm mean diameter and a homogenous, narrow size distribution. At room temperature complexes were stable in 0.9% sodium chloride solution pH 7.4 for 1 h without aggregation. Process parameters such as albumin concentration, incubation time, temperature, pH, order of reagent addition, the presence of bivalent ions and the ionic strength of the complexation medium all influenced the complex size. Confocal laser scanning microscopy showed interactions of a Texas Red labeled cationized albumin with cell membranes of ECV 304 cells and an enhanced endocytic uptake compared to native albumin. The potential for introducing exogeneous DNA into cells was shown using NIH 3T3 fibroblasts. Successful, albeit low reporter gene expression could be achieved in the presence of chloroquine. Under in vitro conditions no toxic effect could be observed. In conclusion, cationized albumin may have promise as a non-toxic vector for gene delivery, especially for DNA vaccination. PMID- 11489559 TI - An in vitro investigation into the effect of glycosaminoglycans on the skin partitioning and deposition of NSAIDs. AB - Recently, Solaraze gel (Bioglan, Herts, UK) a topical hyaluronan (HA)/diclofenac formulation for the treatment of actinic keratosis has received regulatory approval in the US, Canada and Europe for the treatment of actinic keratosis. However, a mechanism of action to explain the topical delivery properties of HA remains to be elucidated. Thus, the aim of this study was to compare the effect of HA with other glycosaminoglycans (chondroitin sulphate (CS), heparin (HP)) and pharmaceutically relevant polysaccharides (sodium carboxymethyl cellulose and pectin) on the dermal partitioning and percutaneous penetration of diclofenac and ibuprofen. The studies demonstrated that HA significantly enhanced the partitioning of both diclofenac and ibuprofen into human skin when compared to an aqueous control, pectin and carboxymethylcellulose (P<0.01). Although the HA vehicle increased the partitioning of both drugs compared to the effects of the other glycosaminoglycans, CS and HP, this difference was not significant (P>0.05). However, the results from the Franz cell diffusion studies showed that HA (1% w/w) significantly enhanced the amount of drug localising within the skin when compared to all of the other polysaccharides (P<0.05). The results suggest that the use of HA as a vehicle excipient offers potential advantages in the dermal delivery and localisation of drugs. PMID- 11489560 TI - A novel approach to the characterization of polar liquids. Part 1: pure liquids. AB - Liquid dosage forms, generally based on aqueous solutions, take an important role in drug administration. The approaches to a theoretical description of solvent and solubility properties have not yet proved completely satisfying. In this work, the Debye equation, which describes well polar and nonpolar molecules in an ideal gas, is extended to liquids. For this purpose, the Debye equation was modified and the term (E(i)/E) was introduced (E(i)=internal electric field, E=applied external electric field). Pure polar and nonpolar solvents were measured between 290.7 and 343.2 K. The values of (E(i)/E) were compared with the correlation factor g of the Kirkwood-Frohlich equation, a measure for molecular pair correlations. For polar solvents, the relationship E(i)/E=m(1/T)+b as a function of temperature T was found. Associating compounds showed negative values of (E(i)/E) with a strong temperature dependency; the latter can be expressed by the slope m. A correlation between the absolute value of m and the corresponding Hildebrand solubility parameter delta could be established. This new approach allows to describe polar hydrogen-bonding liquids and provides a tool for a more rational design of liquid dosage forms. PMID- 11489561 TI - The stability of benzoyl peroxide formulations determined from isothermal microcalorimetric studies. AB - Recent developments in the analysis of microcalorimetric data output allow the possibility of determining both thermodynamic and kinetic parameters for complex reaction systems. Such experiments routinely take around 50 h, hence qualifying for the description rapid. The methods have earlier been applied to a study of the stability of benzoyl peroxide itself in aqueous suspension. This paper reports the results of isothermal microcalorimetric study of the stability of benzoyl peroxide in the presence of a wide range of excipients and in formulated materials. The results are shown to assist in formulation design, are achieved rapidly and are derived from direct experimental study of the complex systems themselves. That is, no ancillary information is required nor are the studies invasive or destructive. PMID- 11489563 TI - Ab externo scleral fixation of the Cionni modified capsular tension ring. AB - The Cionni modified capsular tension ring (CTR) allows for scleral fixation in cases of significant zonular dialysis, providing long-term centration of in-the bag foldable intraocular lenses. Previous techniques of suture placement require placement of the primary incision along the axis of zonular weakness or enlarging and/or distorting the primary incision to attain proper positioning. They also use blind passes of the needle under the iris to approximate the ciliary sulcus. We describe an external closed-system approach for preplacing 10-0 polypropylene sutures in the ciliary sulcus for a Cionni modified CTR under topical anesthesia. This technique does not use blind passes of the suture needle and results in accurate placement of the sutures in the ciliary sulcus under a closed and stable system. PMID- 11489564 TI - Management of posterior capsule rupture during phacoemulsification using the dry technique. AB - To manage posterior capsule rupture during phacoemulsification, we use a dry technique in which all procedures are performed without an irrigation/aspiration system. The dry technique is characterized by (1) continuous viscoelastic injection instead of fluid irrigation to maintain anterior chamber depth with the posterior capsule and vitreous located posteriorly and (2) static removal of most residual lens material by viscoexpression and/or manual small incision extracapsular cataract extraction without aspiration and dynamic water flow. In 16 cases of posterior capsule rupture managed using the dry technique, the residual nucleus and cortex were readily removed with minimum extension of the ruptured area and new vitreous loss. Although large amounts of viscoelastic material (mean 5.8 mL) were required, rapid and stable visual recovery was comparable to that in patients having uneventful surgery. The dry system is a safe and reliable technique for managing posterior capsule rupture during phacoemulsification. PMID- 11489565 TI - Closed-system and open-sky capsulorhexis for combined cataract extraction and corneal transplantation. AB - We describe a triple procedure combining corneal transplantation, cataract extraction, and intraocular lens implantation in which the continuous curvilinear capsulorhexis (CCC) is performed in a closed system when corneal transparency is sufficient or using an open-sky method when corneal transparency is poor. With the closed system, the CCC is performed first followed by trephination and hydroexpulsion of the nucleus. A Caporossi coaxial forceps is used to reduce the corneal incision to 1.2 mm, preventing the need for sutures. Tunnel formation, which could limit the size of trephination, is avoided. In addition, endothelial cell loss at the periphery of the recipient cornea is reduced. This technique was performed in 10 eyes without intraoperative complications. With the open-sky method, the CCC is created while counterpressure is applied to the center of the lens with a large spatula, reducing posterior pressure and thus the risk of capsule tear. This technique was performed in 9 eyes without intraoperative complications. PMID- 11489566 TI - Beam-deflection method of diagnosing impaired vision. AB - We describe a simple method to assign a diagnosis of cataract to patients with obscurely impaired vision as well as to those with mild lens opacities. When the narrowest slit is used in a slitlamp examination, a small beam of light appears on the macula. Routine fundoscopy with the 78.0 diopter lens or a 3-mirror glass is appropriate. If a cataract is present, the beam of light is scattered into several straight lines, distorted lines, or both. This has proved a useful diagnostic tool when the lens appears clear but the patient's vision is impaired, and extensive examinations such as computer tomography or nuclear magnetic resonance tomography for impaired vision may be avoided. The beam-deflection method uses devices that are generally available and can detect cataract in the early stages of development. PMID- 11489567 TI - Improvement in photorefractive corneal laser surgery results using an active eye tracking system. AB - PURPOSE: To study the advantage of modern eye-tracking systems for photorefractive surgery. SETTING: Department of Ophthalmology, University of Zurich, Zurich, Switzerland. METHODS: Photorefractive surgery (photorefractive keratectomy and laser in situ keratomileusis) for myopia and myopic astigmatism was performed in 40 eyes with a commercially available medical excimer laser system. The eyes were selected retrospectively from a larger group of patients treated at 1 clinic. In 20 eyes, the ablation was centered on the entrance pupil using the active, video-based, eye-tracking system (sampling frequency 50 Hz) of the laser. During laser treatment in the nontracker group (20 eyes), the active eye-tracking system was switched off and centration was done manually by the surgeon. Preoperatively and 1 and 3 months after surgery, the patients had a standard ophthalmic examination as well as wavefront analysis by means of a custom-designed wavefront analyzer. RESULTS: After surgery, the visual acuity was significantly better (P <.05) in patients treated with the eye tracker. The increase in coma-like (relative increase factor 0.4) and spherical aberrations (relative increase factor 1.1) was significantly smaller in these patients than in those in the nontracker group (spherical equivalents of 3.9 and 5.1, respectively; P <.05). The refractive outcome, however, was not significantly different in sphere and cylinder. CONCLUSION: The use of active eye tracking appeared to improve the optical and visual outcomes but did not affect the refractive outcome after photorefractive laser surgery. PMID- 11489568 TI - Stability after laser in situ keratomileusis in moderately and extremely myopic eyes. AB - OBJECTIVE: To evaluate the stability, mechanism, and degree of regression following laser in situ keratomileusis (LASIK) in cases with moderate to extreme myopia after 2 years of follow-up. SETTING: Single-center clinical trial. METHODS: Fifty-two eyes of 38 patients were enrolled in the study. One year follow-up was available for 47 eyes of 35 patients and 2 year follow-up for 39 eyes of 27 patients. Eyes were divided into 2 groups based on the level of preoperative myopia: Group 1, moderate to high myopia > or =15.0 diopters (D) (range -7.0 to -15.0 D, n = 24); Group 2, extreme myopia >15.0 D (range -15.3 to 25.8 D, n = 15). Laser in situ keratomileusis was performed using the Chiron Automated Corneal Shaper(R) microkeratome and the Summit OmniMed excimer laser. Manifest spherical equivalent, mean central keratometry, and central corneal thickness (CCT) were measured preoperatively and 12 and 24 months postoperatively. RESULTS: Group 1 exhibited a mild myopic shift (mean -0.07 +/- 0.28 D; P >.2) and a mild increase in keratometry (mean 0.05 +/- 0.46 D; P >.6), with an accompanying increase in CCT (mean 7.5 +/- 12.2 microm; (P =.006) at 24 months. Group 2 displayed a significant myopic shift (mean -0.7 +/- 0.7 D; P =.001) and a significant increase in keratometry (mean 0.4 +/- 0.5 D; P =.01), with a mild increase in CCT (mean 2.4 +/- 9.7 microm; P =.35) at 24 months. Corneal ectasia was evident in 1 eye in the extreme myopia group. CONCLUSION: The refractive effect of myopic LASIK up to -15.0 D remained reasonably stable during the second postoperative year. Significant regression of the refractive effect occurred in eyes with higher levels of myopia (>15.0 D), with the risk of progressive ectasia. Extreme caution is recommended when myopic LASIK is performed in eyes with higher levels of myopia. PMID- 11489569 TI - Laser in situ keratomileusis for residual myopia after primary LASIK. AB - PURPOSE: To analyze the results of secondary laser in situ keratomileusis (LASIK) for residual myopia after primary LASIK. SETTING: Dr. Agarwal's Eye Hospital, Chennai, India. METHODS: A retrospective study of 50 eyes of 29 patients who had a secondary LASIK procedure was carried out. After a mean follow-up of 5.84 months +/- 3.24 (SD) after the primary procedure, the mean myopic residual refraction was -4.30 +/- 1.83 diopters (D). In 10 eyes, the primary corneal flap was lifted by blunt dissection. In 40 eyes, the flap was made with a second cut. The secondary LASIK was performed using the Chiron Technolas Keracor 217 excimer laser and the Automated Corneal Shaper microkeratome (Bausch & Lomb). RESULTS: The mean follow-up was 16.58 +/- 3.06 months. At 12 months, the mean spherical equivalent was -0.45 +/- 0.68 D (P <.05). Thirty-one eyes were emmetropic, 13 eyes were within +/-1.00 D of emmetropia, and 5 eyes were within +/-2.00 D. The mean uncorrected visual acuity improved from 20/80 (range 20/60 to 20/200) to 20/40 (range 20/20 to 20/200) (P <.005). Seventeen eyes gained 1 line of best spectacle-corrected visual acuity; 1 eye lost 1 line because of a decentered ablation with an induced postoperative astigmatism of -3.50 D cylinder. No sight threatening complications such as a free cap, flap irregularity, corneal ectasia, or retinal complication occurred postoperatively. Epithelial ingrowth developed in 5 eyes and corneal thinning, in 1 eye. Three eyes had night glare. CONCLUSION: Secondary LASIK was a safe, stable, and effective method for the treatment of residual myopia after primary LASIK. PMID- 11489570 TI - Treatment of intraocular pressure elevation after photorefractive keratectomy. AB - PURPOSE: To study the effect of timolol maleate, dorzolamide, or a combination of both in post photorefractive keratectomy (PRK) eyes with an elevated intraocular pressure (IOP) after topical steroid administration. SETTING: Refractive Surgery Outpatient Department, 1st Department of Ophthalmology, Semmelweis University, Budapest, Hungary. METHODS: Forty-five patients with elevated IOP were randomly enrolled in 3 groups: Group 1 received timolol maleate 0.5% twice a day; Group 2 received timolol maleate 0.5% twice a day and dorzolamide 2% 3 times a day; and Group 3 received only topical dorzolamide 2% 3 times a day. Intraocular pressure was measured 3 days and 1, 3, and 6 weeks after the antiglaucoma medication was started. RESULTS: The mean preoperative IOP was 15.25 mm Hg +/- 1.28 (SD). Following administration of topical fluorometholone, the IOP increased a mean of 27.39 +/- 2.88 mm Hg. Six weeks after the antiglaucoma therapy was started, the mean IOP reduction was 6.6 mm Hg in Group 1, 8.86 mm Hg in Group 2, and 4.64 mm Hg in Group 3. CONCLUSIONS: A combination therapy of timolol 0.5% and dorzolamide 2% was most effective in treating secondary IOP elevation after PRK. Dorzolamide alone did not adequately control secondary post-PRK IOP elevation. PMID- 11489572 TI - Predicting sulcus size using ocular measurements. AB - PURPOSE: To predict sulcus size using ocular measurements. SETTING: Michel Pop Clinics, Montreal, Quebec, Canada. METHODS: Forty-three eyes were evaluated using several techniques. Ultrasound biomicroscopy (UBM) echograms were taken to measure the anterior chamber depth (ACD), sulcus size, and central corneal thickness. The limbus size was measured with a caliper. Axial length, ACD, and pachymetry were measured by contact ultrasonography. Refraction and corneal power were also evaluated. RESULTS: The coefficient of linear regression was 0.05 between the limbus and the sulcus size (P =.78), 0.76 between ultrasonography and UBM ACD measurements (P <.001), and 0.69 between ultrasonography and UBM pachymetry (P <.001). Paired t tests showed that ultrasound and UBM ACD measurements were not statistically different (P =.70) but that ultrasound and UBM pachymetry measurements were (P <.001). The sulcus versus limbus difference was 0.6 mm for myopia and 0.3 mm for hyperopia. A backward elimination multiple regression performed with all measures to predict sulcus size resulted in the following formula: Sulcus size = 18.9 - 0.023 x sphere + 0.15 x mean keratometry (R = 0.49; P =.005; statistical power = 0.89; standard error of estimate = 0.5 mm). CONCLUSION: Traditional estimation of sulcus size through limbal measurement is inadequate because limbus size alone cannot predict sulcus size. A general formula using the sphere and the mean corneal power can help predict sulcus size. Corneal power was significantly and negatively correlated with sulcus and limbus size as well as sphere. The standard error of sulcus measurement by UBM was 0.4 mm. PMID- 11489571 TI - Erbium:YAG laser emulsification of the cataractous lens. AB - PURPOSE: To evaluate the clinical effectiveness and safety of the erbium:YAG (Er:YAG) laser for cataract extraction surgery. SETTING: Visual Sciences Institute, Madrid, Spain. METHODS: In this prospective randomized study, 65 eyes of 50 patients scheduled for cataract extraction were divided into 2 groups based on the type of lens emulsification: Er:YAG laser (40 eyes) or ultrasound (US) (25 eyes). The laser procedure was converted to the US technique in cases with potential complications. All patients received a foldable posterior chamber intraocular lens. A complete ophthalmologic examination including topographic analysis, pachymetry, and endothelial cell count was performed preoperatively and 1 day, 2 weeks, and 1, 3, 6, and 12 months postoperatively. All patients had fluorescein angiography at 1 month to rule out subclinical cystoid macular edema (CME). RESULTS: There were no significant differences between the techniques in any parameter at any time. In both groups, visual acuity significantly increased and endothelial cell count significantly decreased postoperatively (P <.01). The only serious complication was subclinical CME in 2 patients 1 month after the laser procedure. CONCLUSIONS: Emulsification of the lens using the Er:YAG laser was effective for performing small incision cataract surgery in eyes with soft and medium nuclei. The small ablation zones created can help prevent damage to surrounding ocular structures. The Er:YAG technique used less ablation energy than US emulsification and did not result in thermal injury. PMID- 11489573 TI - Effect of in-the-bag intraocular lens fixation on the prevention of posterior capsule opacification. AB - PURPOSE: To compare the incidence of posterior capsule opacification (PCO) after extracapsular cataract extraction (ECCE) and phacoemulsification and to evaluate the role of posterior chamber intraocular lens (PC IOL) haptic fixation and biomaterial/design in reducing the incidence. SETTING: Postgraduate Institute of Medical Education and Research, Chandigarh, India; Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: This study comprised 278 eyes of 263 patients having ECCE and 318 eyes of 297 patients having phacoemulsification with PC IOL implantation. Posterior capsule opacification leading to a decrease in Snellen visual acuity of 2 or more lines was considered visually significant. The presence of PCO and IOL haptic fixation were evaluated postoperatively using slitlamp biomicroscopy. Haptic position was noted as in-the bag (B-B), 1 haptic in the bag and 1 in the sulcus (bag-sulcus [B-S]), or both haptics out of the bag (sulcus-sulcus [S-S]). In addition, the rate of visually significant PCO was compared among 3 IOL biomaterials: poly(methyl methacrylate), silicone, and hydrophobic acrylic. RESULTS: Visually significant PCO occurred in 42.45% of eyes having ECCE and 19.18% of eyes having phacoemulsification (P <.001, chi-square test) after a mean follow-up of 2.4 years +/- 0.7 (SD). In both groups, visually significant PCO was significantly less in eyes with B-B fixation than in those with B-S or S-S fixation (P <.001). The rate of visually significant PCO in all eyes in the phacoemulsification group with B-B fixation was low (11.90%) and was significantly lower in eyes with a hydrophobic acrylic IOL (2.22%; P <.05, chi-square test). CONCLUSIONS: In-the-bag PC IOL fixation is required to consistently reduce the incidence of PCO. Thorough removal of lens substance, including hydrodissection-assisted cortical cleanup, and in-the-bag PC IOL fixation seem to be the most important factors in reducing PCO, regardless of surgical procedure or IOL type used. Intraocular lens biomaterial and design also help prevent PCO. PMID- 11489574 TI - Posterior capsule opacification and lens epithelial cell layer formation: Hydroview hydrogel versus AcrySof acrylic intraocular lenses. AB - PURPOSE: To quantitatively compare the incidence of visually significant posterior capsule opacification (PCO) and lens epithelial cell (LEC) layer formation on the anterior surface of Hydroview hydrogel and AcrySof acrylic foldable intraocular lenses (IOLs) after implantation. SETTING: Single-surgeon ophthalmology practice, Orange Base Hospital, and Dudley Private Hospital, Orange, New South Wales, Australia. METHODS: This retrospective study comprised 166 eyes of 150 patients (after exclusions) who had cataract extraction and insertion of a foldable IOL in the capsular bag by a single surgeon using a standardized phacoemulsification technique from December 1997 to September 1998. The mean follow-up was 13.1 months (range 6.0 to 23.6 months). The eyes were divided into 2 groups based on the type of IOL implanted: Storz Hydroview H60M (81 eyes) or Alcon AcrySof MA30BA (85 eyes). A neodymium:YAG posterior capsule laser capsulotomy (PC YAG) was performed for an objective decrease in Snellen best corrected visual acuity (BCVA) of more than 1 line, significant visual symptoms, or both. This was used as a measure of visually significant PCO. An Nd:YAG anterior surface clearance (ASC YAG) was done for LEC layer formation anterior to the IOL to better visualize or facilitate treatment of PCO. The rates of PC YAG and ASC YAG after Hydroview and AcrySof IOL implantation were statistically compared. RESULTS: Forty-five eyes (55.6%) in the Hydroview IOL group and 3 eyes (3.5%) in the AcrySof IOL group required a PC YAG; the risk difference was 52.0% (P <.001). An ASC YAG was required in 27 eyes (33.3%) in the Hydroview group and 1 eye (1.2%) in the AcrySof group; the risk difference was 32.2% (P <.001). Survival analysis demonstrated that the only independent predictor of the incidence of PC YAG and ASC YAG over time was IOL type, with the Hydroview IOL group having a statistically significantly higher incidence of both procedures. CONCLUSION: There was a greater incidence of visually significant PCO and LEC layer formation on the anterior surface of Hydroview IOLs than of AcrySof IOLs. PMID- 11489575 TI - Visual acuity and contrast sensitivity in different types of posterior capsule opacification. AB - PURPOSE: To compare the visual acuity (VA) and contrast sensitivity in 2 types of posterior capsule opacification (PCO) in pseudophakic eyes before and after neodymium:YAG (Nd:YAG) capsulotomy and to evaluate vision test results after Nd:YAG capsulotomy. SETTING: Department of Ophthalmology, Taipei Veterans General Hospital, National Yang-Ming University, Taipei, Taiwan. METHODS: Fourteen eyes with fibrosis-type PCO and 15 eyes with Elschnig-pearl-type PCO were enrolled prospectively. Before and 1 week after Nd:YAG capsulotomy, VA and contrast sensitivity were assessed using the illiterate E version of the Bailey-Lovie chart and the Vistech VCTS 6000 chart, respectively. RESULTS: Before capsulotomy, the mean logMAR acuity in the group with Elschnig-pearl-type PCO was 0.47 +/- 0.32 (SD) and in the group with fibrosis-type PCO, 0.17 +/- 0.07. The difference between the 2 groups was significant (P =.002). After capsulotomy, there was no significant between-group difference (P >.05). Before capsulotomy, the contrast sensitivity was significantly worse (P <.01) at all spatial frequencies in the group with pearl-type PCO, especially at 6 cycles per degree. After capsulotomy, there was no significant between-group difference (P >.05) at any spatial frequency. CONCLUSIONS: After cataract surgery, patients with pearl-type PCO had lower VA and contrast sensitivity than those with fibrosis-type PCO. An Nd:YAG capsulotomy improved the VA and contrast sensitivity in patients with both types of PCO. PMID- 11489576 TI - Sharp-edged intraocular lens design as a cause of permanent glare. AB - PURPOSE: To evaluate patients who had implantation of an acrylic intraocular lens (IOL) and reported edge glare that did not lessen with time. SETTING: Hawthorn Eye Clinic, Kew, Victoria, Australia. METHOD: This retrospective study comprised patients who had uneventful phacoemulsification and then reported glare that did not abate after 12 months. RESULTS: Five patients (7 of 322 eyes; 2.2%) who received an AcrySof MA30BA IOL with a 5.5 mm optic and 1 patient (1 of 221 eyes; 0.5%) who received an AcrySof MA60BM IOL with a 6.0 mm optic reported permanent edge glare. Extensive retinal and neurological evaluations in 2 patients showed no deficits in 1 and an abnormal photopic red electroretinogram in the other. Pilocarpine 1% was used in 3 patients and gave no relief of symptoms. Two patients (3 eyes) had IOL explantation with complete resolution of glare symptoms. There was no statistical significance between the 2 types of IOLs. CONCLUSION: Sharp-edged IOL designs can cause permanent intractable glare. PMID- 11489577 TI - Noninvasive Raman spectroscopic identification of intraocular lens material in the living human eye. AB - PURPOSE: To develop a safe noninvasive technique for identifying the material of intraocular lenses (IOLs) implanted in patients. SETTING: Center for Biomedical Engineering and the Department of Ophthalmology and Visual Sciences, University of Texas Medical Branch, Galveston, Texas, USA. METHODS: Raman spectroscopy was used to noninvasively identify the type of IOL implanted after previous cataract surgery in 9 eyes of 6 patients who were legally blind as a result of eye disease. Three IOLs were characterized: poly(methyl methacrylate) (PMMA) (n = 5), acrylic (n = 3), and silicone (n = 1). Confocal Raman spectroscopy was used with a laser power of 95 microW and exposure time of 1 second. RESULTS: Distinct spectral peaks associated with each type of IOL were obtained. These included spectra peaks at 2840 cm(-1), 2946 cm(-1), and 3000 cm(-1) for PMMA; 2917 cm(-1), 2939 cm(-1), and 3055 cm(-1) for acrylic; and 2900 cm(-1), 2961 cm(-1), and 3048 cm(-1) for silicone. The procedure was well accepted by patients, and there were no complications. CONCLUSIONS: The specific Raman spectra of the IOLs allowed for noninvasive determination of IOL material with the use of a safe light dose and an exposure time of 1 second. PMID- 11489578 TI - Anterior lenticonus: histological evaluation and approach for cataract surgery. AB - PURPOSE: To present 2 cases of anterior lenticonus in patients without Alport's syndrome, a surgical technique of cataract extraction in eyes with anterior lenticonus, and histological results of lenticonus specimens obtained intraoperatively. SETTING: From St. Eriks Eye Clinic, Karolinska Institute, Stockholm, Sweden. METHODS: Two otherwise healthy patients presented with anterior lenticonus but no history of Alport's or other pathology. Best corrected visual acuity (BCVA) was decreased. Both patients had cataract extraction by phacoemulsification with intraocular lens implantation under topical anesthesia. Two continuous curvilinear capsulorhexes (CCCs) were created. The entire lenticonus was embedded in formaldehyde buffer 4% for histological analysis. RESULTS: In the first patient, BCVA did not improve postoperatively because of amblyopia. The patient subjectively reported a substantial improvement in visual field clarity. The surgical and postoperative course in the other patient was uneventful. The sections were positive for collagen types IV and VI, and the arrangement of the collagen fibers was highly irregular. CONCLUSION: Anterior lenticonus was detected in patients without Alport's syndrome who were otherwise healthy. A modified 2-step CCC technique can be used to make cataract surgery in such eyes safe and relatively easy. PMID- 11489579 TI - Lens epithelial cell regeneration of a capsule-like structure during postoperative healing in rabbits. AB - PURPOSE: To determine whether lens epithelial cells (LECs) can regenerate the lens capsule during healing after lens extraction and intraocular lens (IOL) implantation. SETTING: Department of Ophthalmology, Wakayama Medical College, Japan. METHODS: Extracapsular lens extraction and IOL implantation were performed in 5 adult albino rabbits. Lens capsules were examined histologically and immunohistochemically 3 and 5 months later. RESULTS: Lens epithelial cells proliferated and regenerated lens fibers within the capsular bag. A multilayered homogenous capsule-like structure was present in the equatorial region. The structures contained type IV collagen but not type I collagen. CONCLUSION: Lens epithelial cells can regenerate lens capsule-like structures during healing after lens extraction. Postoperative LECs without phenotypic conversion to a fibroblastic type may produce this structure. PMID- 11489580 TI - Prevention of posterior capsule opacification by intraoperative single-dose pharmacologic agents. AB - PURPOSE: To determine whether an intraoperative single dose of dexamethasone, diclofenac, ethylenediaminetetraacetic acid (EDTA), a combination of EDTA and RGD peptide (arginine-glycin-aspartic acid sequence), or mitomycin-C (MMC) is a pharmacological means of preventing or reducing the development of posterior capsule opacification (PCO). SETTING: Department of Ophthalmology, Celal Bayar University, School of Medicine, Manisa, and Department of Pathology, Dokur Eylul University, School of Medicine, Izmir, Turkey. METHODS: Fifty-four rabbits were randomly divided into 6 groups. Dexamethasone (4 mg/cc), diclofenac (2.5 mg/cc), EDTA (8 mg/cc), a combination of EDTA and RGD peptide (2.5 mg/cc), or MMC (0.04 mg/cc) was given, 0.1 cc by hydrodissection and 0.9 cc into the capsular bag after phacoemulsification. The sixth group served as a control group. After 3 months, the PCO was graded clinically and the proliferation of lens epithelial cells (LECs) was evaluated histologically. RESULTS: The drugs were significantly effective in preventing PCO compared with the control (P <.005). Dexamethasone had a weaker effect than the other drugs. In histological analysis, although monolayer LECs in the dexamethasone and diclofenac groups were observed, there was no proliferative activity on the posterior capsules in the EDTA, EDTA+RGD, and MMC groups in contrast to the multilayer cells in the control. CONCLUSIONS: Intraoperative single-dose application of EDTA, EDTA+RGD peptide combination, and MMC significantly prevented the development of PCO in rabbit eyes. Diclofenac was less effective but also reduced PCO. Although dexamethasone did not prevent the proliferation of LECs, it decreased PCO clinically. PMID- 11489581 TI - Comparison of Scheimpflug images of posterior capsule opacification and histological findings in rabbits and humans. AB - PURPOSE: To compare the posterior capsule opacification in Scheimpflug photographic images produced by an electronic anterior eye segment analysis system with the histopathological findings in rabbits and humans. SETTING: Department of Ophthalmology, Wakayama Medical College, Japan. METHODS: Opacified posterior capsules were photographed using the EAS-1000 system (Nidek) and were then extracted during vitreous surgery for proliferative diabetic retinopathy or proliferative vitreoretinopathy in 2 patients. In rabbits, phacoemulsification and aspiration (PEA) with intraocular lens (IOL) implantation was performed. The IOL was implanted in the bag or in the sulcus. After intervals of healing, the posterior capsule was photographed with the EAS-1000 and the animals were then killed. In both clinical and experimental specimens, the posterior capsule was processed for light microscopic histology and immunohistochemistry. RESULTS: Opacified human capsules were well imaged by the EAS-1000. Histology showed that lens epithelial cells proliferated with and without an accumulation of extracellular matrix. Details such as rolling of the capsulotomy edge were seen well. Regenerated lens fibers of Soemmering's ring were seen as a mass within the capsule. In the rabbit model, Scheimpflug images accurately represented the capsules as they appeared histologically. CONCLUSION: The EAS-1000 system provided faithful, relatively high-resolution images that corresponded to the histologic findings in the posterior capsules after PEA-IOL surgery in humans and rabbits. PMID- 11489582 TI - Optics of aberroscopy and super vision. AB - This paper (1) reviews the fundamental limits to visual performance imposed by optical imaging and photoreceptor sampling to determine the limits to the potential gains offered by ideal corrections; (2) examines the predicted losses in vision induced by chromatic aberration, phase shifts, typical ocular aberrations, and the gains possible by correcting the monochromatic aberrations of the eye; (3) discusses the principles of aberration measurement in the eye; and (4) presents methods for measuring and classifying monochromatic aberrations of the eye. PMID- 11489583 TI - Late partial dislocation of a laser in situ keratomileusis flap. AB - We report an apparently atraumatic asymptomatic flap dislocation 4 months after uneventful laser in situ keratomileusis (LASIK) in the right eye of a 43-year-old woman. The patient developed partial dislocation of the LASIK flap during the week after the 4 month examination. The LASIK flap was subsequently lifted to perform an enhancement, and the postenhancement course has been unremarkable. This case illustrates the potential susceptibility of LASIK flaps to dislocation either spontaneously or, more likely, after presumed minor trauma as late as 4 months after the original procedure. PMID- 11489584 TI - Late traumatic flap dislocation after laser in situ keratomileusis. AB - Three patients who had laser in situ keratomileusis (LASIK) experienced severe direct corneal trauma 3 to 38 months postoperatively. Flap dislocation resulted in all 3 patients. Presentation following trauma ranged from a few hours to 9 days. All patients were successfully managed by surgical flap repositioning, and all maintained a best corrected visual acuity of 20/20. A literature review of traumatic flap dislocation cases and the most recent methods of diagnosis and management are included. PMID- 11489585 TI - Bilateral severe keratoconus after laser in situ keratomileusis in a patient with forme fruste keratoconus. AB - We report a case of bilateral keratoconus after laser in situ keratomileusis (LASIK). Before surgery, the patient had a forme fruste keratoconus, which evolved rapidly to a severe form of keratoconus in the months following LASIK. From this case, we conclude that forme fruste keratoconus is a contraindication to LASIK. PMID- 11489586 TI - Contact lens fitting in a patient with keratectasia after laser in situ keratomileusis. AB - We present a case of unilateral iatrogenic keratectasia developing 15 months after bilateral laser in situ keratomileusis using a broad-beam excimer laser (Bausch & Lomb Keracor 116) to treat -3.5 -1.5 x 85 diopters of myopia. Preoperative pachymetry in the eye measured 450 microm without topographical changes suggesting keratoconus or forme fruste keratoconus. Contact lens fitting to provide 20/25 visual acuity is described. PMID- 11489587 TI - Late-onset interface keratitis after uneventful laser in situ keratomileusis. AB - Although there have been many proposed etiologies for interface inflammation after laser in situ keratomileusis, the causative factor(s) remains an enigma. This case of late-onset interface inflammation suggests that the many previously suspected interface contaminants from the surgical environment were not involved. Transformation of a previously inert material to an inflammatory stimulus is proposed as a potential cause; however, a sample of inflammatory focus was not obtained because of the immediate and complete response to frequent topical steroid drops. PMID- 11489588 TI - Diffuse lamellar keratitis associated with iritis 10 months after laser in situ keratomileusis. AB - A 35-year-old man developed diffuse lamellar keratitis (DLK) 10 months after laser in situ keratomileusis (LASIK). The DLK was associated with acute iritis, not with manipulation of the LASIK flap or epithelium. This case supports the hypothesis that DLK is a nonspecific inflammatory response of the cornea rather than a specific agent causing the syndrome. PMID- 11489589 TI - Corneal opacity after repeated photorefractive keratectomy. AB - Corneal opacity developed in an eye that had photorefractive keratectomy (PRK) with a 193 nm excimer laser 5 times over 3 years. Six months after the last PRK, a partial penetrating keratoplasty was performed. The cornea was stained and immunohistochemically evaluated for collagen types. Light microscopy showed thickening of epithelial layers, proliferation of subepithelial fibroblasts, and the absence of Bowman's membrane. Transmission electron microscopy showed irregular collagen lamellae and electron-dense deposits adjacent to keratocytes. The staining was positive for Alcian blue, and immunohistochemistry was positive for type IV and VI collagen. This case suggests that corneal opacity after repeated PRK is the result of deposits of type IV and VI collagen and acidic mucoprotein in the extracellular matrix. PMID- 11489590 TI - Rupture of a radial keratotomy incision after 11 years during clear corneal phacoemulsification. AB - We report a case of rupture of a radial keratotomy (RK) incision that occurred during clear corneal phacoemulsification 11 years after the initial surgery. The RK was done in both eyes for correction of high myopia (>8.0 diopters). This was followed by 2 enhancement procedures at 6 month intervals. The patient presented with diminished vision in both eyes. The diagnosis was nuclear cataract in the right eye, and clear corneal phacoemulsification was done. The intraoperative and postoperative courses were uneventful. Nine months later, clear corneal temporal phacoemulsification was done in the left eye. During surgery, 1 of the radial incisions opened to one third its length. The wound was sutured, and the procedure was completed uneventfully. One month later, best corrected visual acuity was 20/20. PMID- 11489591 TI - Fetal effects of cocaine: an updated meta-analysis. AB - BACKGROUND: A very large number of women in the reproductive age group consume cocaine, leading to grave concerns regarding the long term health of millions of children after in utero exposure. The results of controlled studies have been contradictory, leading to confusion, and, possible, misinformation and misperception of teratogenic risk. OBJECTIVE: To systematically review available data on pregnancy outcome when the mother consumed cocaine. METHODS: A meta analysis of all epidemiologic studies based on a priori criteria was conducted. Comparisons of adverse events in subgroups of exposed vs. unexposed children were performed. Analyses were based on several exposure groups: mainly cocaine, cocaine plus polydrug, polydrug but no cocaine, and drug free. RESULTS: Thirty three studies met our inclusion criteria. For all end points of interest (rates of major malformations, low birth weight, prematurity, placental abruption, premature rupture of membrane [PROM], and mean birth weight, length and head circumference), cocaine-exposed infants had higher risks than children of women not exposed to any drug. However, most of these adverse effects were nullified when cocaine exposed children were compared to children exposed to polydrug but no cocaine. Only the risk of placental abruption and premature rupture of membranes were statistically associated with cocaine use itself. CONCLUSIONS: Many of the perinatal adverse effects commonly attributed to cocaine may be caused by the multiple confounders that can occur in a cocaine using mother. Only the risk for placental abruption and PROM could be statistically related to cocaine. For other adverse effects, additional studies will be needed to ensure adequate statistical power. PMID- 11489592 TI - Nonsteroidal anti-inflammatory drugs in early pregnancy. AB - A study was performed of congenital malformations in infants whose mothers used nonsteroidal anti-inflammatory drugs (NSAIDs) in early pregnancy. Data were obtained from an ongoing prospective recording of drug use during the first trimester. During the period July 1, 1995 through December 31, 1998, 2557 infants were born to women who reported the use of NSAIDs in early pregnancy. The OR (after consideration of maternal age, parity, and smoking habits) for any congenital malformation was 1.04 (95%CI 0.84-1.29), but the OR for cardiac defects reported to the Medical Birth Registry was 1.86 (1.32-2.62) based on 36 instances, and for orofacial clefts 2.61 (1.01-6.78) based on only six instances. By using other information sources, another four infants with cardiac defects were identified. There was no drug specificity for cardiac defects but among six mothers of infants with orofacial clefts, five had used naproxen. PMID- 11489593 TI - Effect of dioxin on ovarian function in the cynomolgus macaque (M. fascicularis). AB - Ovarian function was evaluated in mature female cynomolgus macaques 443 to 625 days following a single oral exposure (1, 2, or 4 microg/kg BW) to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). Urinary estrone conjugates (E1C), pregnanediol-3-glucuronide (PdG), and follicle stimulating hormone (FSH) were measured. Three of four animals in the high dose group had no evidence of menstrual cycles while animals in the low and medium dose groups plus one from the high dose group had cycles that were similar to those of control animals. The noncycling animals had baseline E(1)C concentrations without ovulatory midcycle peaks and monotonic PdG profiles. Mean FSH concentrations during the midfollicular phase of the medium dose group and during the entire cycle of the high dose group were elevated compared to those of the control group and the endometria of the noncycling animals were inactive. These data demonstrate that a single exposure of 4 microg/kg BW TCDD leads to long-term adverse effects on ovarian function in primates. PMID- 11489594 TI - Lead accumulation in the mouse ovary after treatment-induced follicular atresia. AB - Although the main target of lead (Pb) toxicity is the red blood cell, Pb associated changes in the nervous system, the kidney, and the reproductive system have also been described. The few Pb studies conducted on females revealed mostly miscarriages, premature delivery, and infant mortality in humans and animals. This study was done to correlate Pb accumulation in the ovary with damage to folliculogenesis. Pb burden was assayed by atomic absorption spectrometry in bone, liver, adrenal glands, ovary, and fetuses taken from mice exposed according to 2 protocols: intraperitoneal (i.p.) injection of Pb(NO(3))(2) 10 mg/kg/day for 15 days or 10 mg/kg/week for 15 weeks. Ovaries were examined histologically. Pb accumulation in the various soft tissues of acutely exposed mice was similar, and significantly higher than in the organs of chronically exposed mice. A low Pb concentration in the ovary caused dysfunction of folliculogenesis, with fewer primordial follicles and an increase in atretic antral follicles. PMID- 11489595 TI - Phenanthrenequinone disrupts progesterone production in rat luteal cells. AB - The ability of the environmental contaminant phenanthrene (PH) and its photooxidized product phenanthrenequinone (PHQ) to disrupt progesterone secretion was examined in a model system of in vitro suspensions of luteal cells from the rat. Treatment with PHQ dramatically inhibited luteinizing hormone (LH) stimulated progesterone secretion. PHQ also generated a significant increase in reactive oxygen species (ROS). In the absence of LH, however, PHQ stimulated a small increase in basal progesterone secretion. The parent compound, PH, did not alter progesterone or ROS release. Since there is evidence that PHQ lowers the activity of nitric oxide synthase (NOS) and that nitric oxide (NO) affects progesterone production, we examined the response to the NOS inhibitors N monomethyl-L-arginine, Zn protoporphyrin-9, and aminoguanidine in luteal cells. However, there was no effect of these agents on LH stimulated progesterone secretion. These results indicated that PHQ is a potent disrupter of progesterone secretion and should perhaps be considered in assessing the risk of PH to humans. PMID- 11489596 TI - Reproductive effects in male and female rats of neonatal exposure to genistein. AB - Sprague-Dawley rats were administered genistein orally at doses of 12.5, 25, 50, or 100 mg/kg on postnatal days 1 through 5 to examine its effects on reproductive function after puberty. In addition, preputial separation and vaginal opening as endpoints of sexual maturation, estrous cycling, sperm count, serum testosterone concentration, and histopathologic changes of reproductive organs of male and female rats were examined. Body weights of male and female rats exposed to genistein at any dose level examined were lower than those of controls. Timing of preputial separation in males and timing of vaginal opening were not affected by genistein treatment. The number of females showing estrous cycle irregularities was increased by genistein treatment. The fertility of female rats exposed neonatally to genistein at 100 mg/kg was disrupted, while neonatal exposure to genistein did not affect male fertility. Neither sperm counts nor serum testosterone concentration were changed by neonatal exposure to genistein. Female rats exposed neonatally to genistein at 100 mg/kg showed histopathologic changes in the ovaries and uterus, while male rats showed no histopathologic alterations in the gonads. The results of this study indicate that early neonatal exposure to genistein caused dysfunction of postpubertal reproductive performance as well as abnormal development of gonads in female but not in male rats. PMID- 11489597 TI - Developmental toxicity evaluation of 1,2,3,4-butanetetracarboxylic acid in Sprague Dawley (CD) rats. AB - 1,2,3,4-butanetetracarboxylic acid (BTCA), proposed as a formaldehyde substitute in the treatment of permanent press fabrics, was evaluated for developmental toxicity. Timed-mated CD rats (25 per group) received BTCA 250, 500, or 1000 mg/kg/day or vehicle (deionized/distilled water) by gavage on gestational days (gd) 6 through 19. Maternal feed and water consumption, body weight, and clinical signs were monitored throughout gestation. At termination (gd 20), confirmed pregnant females (21 to 25 per group) were evaluated for clinical status and gestational outcome; live fetuses were examined for external, visceral, and skeletal malformations. One maternal death, reduced body weight, and reduced weight gain were noted at the high dose; confirmed pregnancy rates were 84 to 100% for each group. There were no treatment-related effects on fetal growth, survival, or morphologic development. The maternal toxicity NOAEL and LOAEL are 500 and 1000 mg/kg/day, respectively. The developmental toxicity NOAEL is > or = 1000 mg/kg/day, and the LOAEL was not established in this study. PMID- 11489598 TI - Antifungal triazoles induce malformations in vitro. AB - Triazole-derivatives are antimycotics used in agriculture as well as in clinical and veterinary therapy. The aim of the present work is the in vitro comparative study of the teratogenic activity of triazole (the parental compound), flusilazole (an agricultural triazole mono-derivative fungicide), and fluconazole (a clinically used bis-triazole derivative). Rat embryos, 9.5 days old (1 to 3 somites) were exposed in vitro to triazole 500 to 5000 microM, flusilazole 3.125 to 250 microM, or fluconazole 62.5 to 500 microM. After 48 h in culture, the embryos were morphologically examined and processed for histologic and biochemical analysis. Flusilazole and fluconazole showed similar teratogenic effects (abnormalities at the branchial apparatus level and cell death at the level of the branchial mesenchyme) at concentration levels of 6.25 microM and higher for flusilazole and of 125 microM and higher for fluconazole. By contrast, only slight developmental retardation and blood discoloration were observed at the highest concentrations of triazole, suggesting no teratogenic activity for the triazole group. PMID- 11489599 TI - (14)C methanol incorporation into DNA and proteins of organogenesis stage mouse embryos in vitro. AB - Methanol (MeOH), a widely used industrial solvent and alternative motor fuel, has been shown to be mutagenic and teratogenic. We have demonstrated that methanol is teratogenic in mice in vivo and causes dysmorphogenesis in cultured organogenesis stage mouse embryos. Although MeOH is a product of endogenous metabolism in the gut and can be found in humans following consumption of various foods, elevated levels of methanol could lead to methylation of cellular macromolecules. DNA methylation has been demonstrated to suppress transcription of fetal genes and may also play an important role in genetic imprinting. Embryonal proteins are also potential targets for methanol-induced methylation. We investigated the potential of administered methanol to incorporate into and/or alter the methylation of embryonal DNA or to affect specific protein methylation. Gestational day 8 CD-1 mouse embryos were grown for 24 h in culture medium (CM) with 0, 4, or 8 mg MeOH + 20 microCi (14)C-MeOH/mL. At the end of the culture period, yolk sacs and embryos were separated for each treatment group. The DNA was purified by cesium chloride gradient centrifugation in the presence of ethidium bromide and (14)C incorporation was determined. Methylation of a selected gene, Hoxc-8, was assessed by using methylation-specific restriction enzymes. The (14)C activity was found superimposed over the DNA-containing fraction, indicating incorporation. DNA from embryos treated with 4 mg MeOH/mL CM gave the highest incorporation of (14)C-MeOH (8 mg/mL was growth inhibiting). Methylation of Hoxc-8 appeared to be increased in embryos treated with 4 mg MeOH/mL CM, but not in embryos treated with 8 mg MeOH/mL. Lack of incorporation of methylation at the higher concentration may be due to the failure of embryos to grow at this concentration of MeOH. The incorporation of (14)C-MeOH into embryo proteins was investigated by polyacrylamide gel electrophoresis (PAGE) and autoradiography. Incorporation of (14)C-MeOH into specific proteins was observed but the labeling specificity was not methanol dose-related. These results indicate that methyl groups from (14)C-MeOH are incorporated into mouse embryo DNA and protein. Our results further suggest that methanol exposure may increase genomic methylation under certain conditions which could lead to altered gene expression. PMID- 11489600 TI - Strain sensitivity differences in the Hershberger assay. AB - The Hershberger assay is a test method for detecting androgenic or antiandrogenic properties based on alterations in the weights of accessory sex organs in castrate male animals. We performed this study to examine strain sensitivity differences in the Hershberger assay. Flutamide (FLU) at a dose of 3.2 mg/kg was administered to castrated F344, SD, or Wistar rats, in addition to testosterone propionate (TP) administered at a dose of 0.4 mg/kg. Although FLU significantly attenuated the TP-induced increase in glans penis weight in SD and Wistar rats, this attenuation was not observed in F344 rats. Statistical analysis showed differences among the strains in all sex accessory organ weights. The interaction in the ventral prostate, seminal vesicle, and glans penis weights was significant between SD and F344 rats, and between Wistar and F344 rats, but not between SD and Wistar rats. F344 rats were less suitable than SD or Wistar rats for detecting FLU-induced changes. PMID- 11489603 TI - Neural activity associated with the realization of a delayed intention. AB - This study examines neural activity associated with the realization of a delayed intention within the context of the noticing+search model of prospective memory (PM) using event-related brain potentials (ERPs). The noticing+search model proposes that PM is supported by two related processes, noticing (the detection of a PM cue in the environment) and search (the retrieval of an intention from memory). In two experiments participants performed a PM task that permitted the dissociation of the noticing and search processes. Noticing was associated a phasic negativity over the occipital-parietal region (N320) and search was associated with a sustained modulation (slow-wave) reflecting a negativity over the right frontal region and a broadly distributed positivity over the parietal region. The amplitude of the N320 was greater when the PM cue was associated with an intention than when the cue was irrelevant to task performance, leading to the proposal that noticing may be accomplished through the attentional modulation of neural systems which support processing of the defining features of the PM cue. The topography of the slow-wave resembled that of modulations of the ERP associated with the recollection of information in studies of retrospective memory leading to the suggestion that similar neural processes may support the recovery of information from memory in both prospective and retrospective memory tasks. PMID- 11489604 TI - Electrophysiological correlates of memory encoding are task-dependent. AB - Event-related potentials (ERPs) were used to investigate whether the neural correlates of successful episodic encoding differ according to the nature of the study task. At study, 16 subjects were cued to make either animacy or alphabetic decisions about visually presented words. A recognition memory test with confidence judgements followed after a delay of 30 min. For the animacy task, words that were subsequently confidently recognised were associated with a positive-going ERP modulation. By contrast, for the alphabetic task, confident recognition was associated with a negative-going ERP modulation. Both types of subsequent memory effects started shortly after word onset. These findings suggest that the neural correlates of memory encoding differ qualitatively, rather than quantitatively, according to the nature of the study task. Episodic encoding thus seems to be supported by multiple, task-specific, neural systems. The early onset of these memory effects suggests that episodic encoding can be facilitated by processes that start before the onset of the to-be-encoded item. PMID- 11489605 TI - Neural activities during Wisconsin Card Sorting Test--MEG observation. AB - The present study recorded activities of magnetoencephalography (MEG) to the presentation of cards, and to the presentation of feedback signals in 12 normal subjects while they performed the Wisconsin Card Sorting Test (WCST), to observe temporal and spatial processing during the task. The MEG responses were compared between two different conditions in the presentation both of cards and of feedback signals: the cards proceeded by the first wrong [W1st(C)] and by the 4th correct feedback signals [C4th(C)]; and the feedback of the first wrong [W1st(FB)] and the 4th correct signals [C4th(FB)]. A multi-dipole model, brain electric source analysis (BESA), was used to explore the dipole sources responsible for the MEG activities. We found that MEG activity differences between the W1st(C) and the C4th(C) condition occurred in the period of 190-220 ms (M190 and M200), and 300-440 ms (M300 and M370) mainly at the supramarginal gyrus, the dorsolateral prefrontal, and the middle and inferior frontal gyrus. MEG differences between the W1st(FB) and the C4th(FB) condition occurred 460-640 ms (M460) after the presentation of the feedback signals, with the activation of the dorsolateral prefrontal cortex and the middle frontal cortex. No significant location differences were found between the frontal responses (M370) of the W1st(C) and M460 of the W1st(FB). Our results proved that the WCST task activates a broad frontal area and the parieto-frontal network across time streaming. Both shifting attention to the wrong feedback and enhanced visual working memory to the sorting shifting condition of the card presentation occur in the same areas at different time points. PMID- 11489606 TI - Theta synchronization during episodic retrieval: neural correlates of conscious awareness. AB - The neural correlates of conscious awareness during successful memory retrieval were examined. In a recognition test, subjects indicated whether they consciously recalled the event in which a word was earlier presented (Remembering), or whether they recognized it on the basis that it was familiar in the absence of recollection (Knowing). An early EEG synchronization in the theta band predicted knowing, and a later remembering. Moreover, early and late event-related potentials were also found to predict knowing and remembering, respectively. The results indicate that the temporal dynamics of theta synchronization are related to the particular conscious experiences associated with memory retrieval. PMID- 11489607 TI - Automatic discriminative sensitivity inside temporal window of sensory memory as a function of time. AB - Neural representation of preceding sound-patterns stored in the human brain, as reflected by mismatch negativity (MMN) related to the automatic discriminative process, is restricted to a duration of 160-170 ms due to the short form of auditory sensory memory termed the temporal window of integration (TWI). To examine the temporal uniformity of deviation-sensitivity inside TWI of sensory memory, magnetic MMN (MMNm) responses were measured with a dual 37-channel magnetometer for complex sounds of 170 ms duration containing an omitted (silent) segment. Frequent standard stimuli (probability of 80%) consisted of five tone segments. Deviant stimuli were different from standard stimuli in that one of four segments was occasionally (probability of 5%) omitted and replaced by a silent segment. The stimulus duration of 170 ms was intended to correspond to the postulated duration of TWI. When the silent segment occurred later in deviant stimulus, the MMNm peak amplitude was attenuated and MMNm peak latency, measured from the onset of each silent segment, was delayed. Thus, automatic deviation detection sensitivity declines nonlinearly toward the end of TWI in auditory sensory memory. In the second experiment, two types of deviant stimuli, which differed from each other only in the period after the occurrence of the silent segment, elicited MMNm with the same peak latency but with a different peak amplitude. Thus, mismatch process is triggered at the moment of change but still lasts after the detection of deviation. In other words, both standard and deviant stimuli are treated as a unitary event within a TWI. PMID- 11489608 TI - Physiological evidence of gender differences in word recognition: a magnetoencephalographic (MEG) study. AB - Magnetic field recordings were made in order to describe brain processes during a word recognition experiment. We investigated 26 healthy young subjects (14 females) and focused on gender differences related to recognition performance and brain activity. From about 200 ms to 350 ms after word onset the event-related field (ERF) patterns differed significantly between women and men, although the mean recognition performances did not. Differences were due to different strengths of activation as well as due to the involvement of different neural structures as underlined with statistical analysis. We interpret that our physiological findings demonstrate that different mental strategies are used for correct word recognition in the brains of women and men as assessed with magnetoencephalography (MEG). Our data might be linked to previous findings about the hemispheric asymmetry in male subjects (left lateralized) compared to women in whom both hemispheres seem to be equally involved in word processing. PMID- 11489609 TI - Differences in EEG current density related to intelligence. AB - Differences in current density between high intelligent (IQ=127), and low intelligent individuals (IQ=87), while solving two oddball tasks (auditive and visual) were analyzed with low resolution brain electromagnetic tomography (LORETA). In highly intelligent individuals a decrease in the volume of activated cortical gray matter between the P300 onset and the P300 peak amplitude was observed. The EEG of low intelligent individuals showed a reverse pattern of cortical activity. In the auditive oddball task the decrease in the activated cortical volume in high intelligent individuals was accompanied by an increase in current density, and a more left hemispheric source location at maximum current density. The results suggest that high intelligent individuals more efficiently distributed their cognitive resources needed to cope with the oddball tasks. PMID- 11489610 TI - An outline of brain function. AB - An outline of how the brain may compute is proposed. In the cerebral cortex memories are stored through long-term potentiation at synapses from layer 1 cortical inputs (representing contexts) on layer 2/3 pyramidal cells linked with the thalamus in a cortico-thalamic (CT) unit. The signals which are memorized are the layer 3 inputs from the thalamus or other cortical areas. Signals are memorized (and later recalled) at the gamma frequency. A conscious thought comprises the outputs of layer 5 cells in CT units in different cortical regions firing in synchrony through the contribution of oscillatory thalamic and cortical inputs. This cortical output influences sub-cortical areas to cause or participate in a movement. Cerebral cortical outputs may be stored in the cerebellum and generated later in a particular context by the basal ganglia and cerebellum. Thus the brain may either generate 'conscious' outputs using the cerebral cortex or 'automatic' outputs using the basal ganglia and cerebellum. When contexts are recognized by the basal ganglia it permits outputs stored in the cerebellum to commence and in this way the basal ganglia can control complex sequences of outputs or movements. Working memory involves the prefrontal cortex using similarly the basal ganglia and cerebellum. The hippocampus has a role in the storage and recall of cortical outputs by providing unique layer 1 contexts to all the CT loops in different cortical areas in a conscious thought. With further recall of the thought new layer 1 contexts may become associated with the CT loops enabling recall without the hippocampal input. PMID- 11489611 TI - Visual activation of frontal cortex: segregation from occipital activity. AB - Studies in primates have found visually responsive neurons that are distributed beyond cortical areas typically described as directly involved in vision. Among these areas are premotor cortex, supplementary motor area, dorsolateral prefrontal cortex and frontal eye fields. Given these findings, visual stimulation would be expected to result in activation of human frontal cortex. However, few human studies have described sensory activations in frontal regions in response to simple visual stimulation. Such studies have classically described event-related potential (ERP) components over occipital regions. The present study sought to further characterize the spatiotemporal dynamics of visually evoked electrocortical responses elicited by simple visual stimuli using scalp current density measures derived from high-density ERP recordings, with particular emphasis on the distribution of stimulus-related activity over frontal cortex. Hemiretinal stimuli were viewed passively and during a simple ipsi- or contramanual (RT) task. The motor requirement was included to investigate the effects of response preparation on premovement frontal activations. The results indicate early frontocentral activation, particularly over the right hemisphere (peak magnitude 124-148 ms) that is independent of input visual field or motor response requirement, and that is clearly separate in timecourse from the posterior responses elicited by visual input. These findings are in accord with the multiplicity of visual inputs to frontal cortex and are discussed in terms of frontal lobe functions as may be required in these tasks. PMID- 11489612 TI - Frontal-parietal activation differences observed before the execution of remembered saccades: an event-related potentials study. AB - Healthy subjects performed saccadic eye movements in one memory (MEM) and two delay tasks (delay, DEL and modified delay, M-DEL) while we recorded scalp event related potentials (ERPs) from 25 electrode sites. In the MEM task the subjects were instructed to retain in memory the location of a visual target for a delay of 1-6 s and then perform a remembered saccade at the go signal. In the DEL task the target remained on until movement completion and in the M-DEL task the target, that was visible during the delay period, disappeared synchronously with the go signal. A reduction in response latency and an increase in the percentage of dysmetric movements were observed for the MEM task compared to the two delay tasks. An increased ERP activity at the central-frontal electrode sites compared to the parietal sites was significant only for the MEM task early on during the delay period (500-1000 ms). During the period preceding the onset of the saccade, a parietal increase of activity was observed for all tasks. Furthermore the activity was smaller for the frontal compared to the parietal areas only for the memory task thus indicating a near reversal of the previous pattern of activity observed during the early delay period. This specific activation pattern of frontal and parietal areas, observed for the MEM task only, requires further investigation focusing on the temporal pattern of activation of large brain areas involved in working memory processing. PMID- 11489613 TI - Activated brain regions in musicians during an ensemble: a PET study. AB - As in visual processing, we speculated that, in music processing, different brain regions would activate according to the mode of music listening. Using motets by a famous composer, we studied changes in regional cerebral blood flow (rCBF) with positron emission tomography associated with concentrating on the alto-part within the harmony (alto-part-listening condition) compared to listening to the harmony as a whole (harmony-listening condition). The alto-part-listening condition was associated with bilateral increases of rCBF in superior parietal lobules, precunei, premotor areas and orbital frontal cortices. Superior parietal lobules are likely to be responsible for auditory selective attention to the alto part within the harmony and the analysis of tone pitch on a mental score. The precuneus possibly participated in writing tones of the alto part on a mental score. Based on our findings, we propose that both auditory selective attention and analytic processing play an important role in concentrating on a certain vocal part within a harmony. During the harmony-listening condition, temporal poles, the anterior portion of the cingulate gyrus, occipital cortex and the medial surface of the cerebellum were bilaterally activated. Further studies are necessary to clarify the difference in music processing between musicians and nonmusicians. PMID- 11489614 TI - Effect of ketamine on the neuromagnetic mismatch field in healthy humans. AB - Mismatch negativity (MMN) is a component of the auditory evoked event-related potentials (ERP) that assesses automatic sound change detection and is disturbed in schizophrenic patients. Animal experimental evidence has linked the generation of MMN to the N-methyl-D-aspartate (NMDA) receptor. We investigated the neuromagnetic mismatch field (MMF) in healthy volunteers before and after intravenous application of a subanesthetic dose of the NMDA receptor antagonist ketamine (0.3 mg/kg). Ketamine had a significant influence on latency and dipole moment of the MMF, whereas the N100m latency of the standard tone was not prolonged and its dipole moment remained stable. Our results suggest that ketamine interferes with aspects of preattentive information processing and is in line with the view that disturbed NMDA receptor function may mediate the deficient auditory mismatch response in patients with schizophrenia. PMID- 11489615 TI - Feedback-dependent modulation of isometric force control: an EEG study in visuomotor integration. AB - The primary purpose of this investigation was to examine the cortical mechanisms underlying visuomotor integration in an experiment directly manipulating visual feedback (control-signal gain) as participants executed a grasping task. This was accomplished by assessing human electroencephalograms in both time and frequency domains and relating these measures to the performance accuracy of isometric force control. The basic experimental manipulation consisted of subjects controlling a grip dynamometer and the subsequent force trace displayed on a computer monitor at various magnitudes of force output and control-signal gain. Several findings from this study were of interest. First, the effects of control signal gain and its interplay with the magnitude of force were most evident across the parietal and frontocentral electrode locations--areas specifically related to multi-modal sensory evaluation (parietal lobe) and higher-order movement control (supplementary and mesial premotor areas). Second, electroencephalography (EEG) measures in the time domain, i.e., slow-wave potentials, were sensitive to control-signal gain only during the ramp phase of force production (period of reaching the target force), not the static phase (period of maintaining the target force level). Third, EEG measures within the frequency domain (event-related desynchronization), unlike the slow-wave potential measures, were sensitive to control-signal gain during the static phase of force production--a sensitivity that was directly related to improvements in the accuracy of isometric force control. The findings of this investigation are described in relation to the existent literature on human visuomotor integration with special attention paid to the distinct spatial and temporal electrocortical patterns exhibited under varying degrees of visual feedback and magnitudes of force output during grasping. PMID- 11489616 TI - Simultaneously active pre-attentive representations of local and global rules for sound sequences in the human brain. AB - Regular sequences of sounds (i.e., non-random) can usually be described by several, equally valid rules. Rules allowing extrapolation from one sound to the next are termed local rules, those that define relations between temporally non adjacent sounds are termed global rules. The aim of the present study was to determine whether both local and global rules can be simultaneously extracted from a sound sequence even when attention is directed away from the auditory stimuli. The pre-attentive representation of a sequence of two alternating tones (differing only in frequency) was investigated using the mismatch negativity (MMN) auditory event-related potential. Both local- and global-rule violations of tone alternation elicited the MMN component while subjects ignored the auditory stimuli. This finding suggests that (a) pre-attentive auditory processes can extract both local and global rules from sound sequences, and (b) that several regularity representations of a sound sequence are simultaneously maintained during the pre-attentive phase of auditory stimulus processing. PMID- 11489617 TI - Attention-dependent suppression of distracter visual input can be cross-modally cued as indexed by anticipatory parieto-occipital alpha-band oscillations. AB - Recent studies show that in addition to enhancing neural processing for attentionally relevant stimuli, selective attention also operates by suppressing the processing of distracter stimuli. When subjects are pre-cued to selectively deploy attention during voluntary (endogenous) attentional tasks, these mechanisms can be set up in advance of actual stimulus processing. That is, the brain can be placed in a biased attentional state. Two recent cueing studies have provided evidence for the deployment of such biased attentional states [J.J. Foxe, G.V. Simpson, S.P. Ahlfors, Neuroreport 9 (1998) 3929-3933; M.S. Worden, J.J. Foxe, N. Wang, G.V. Simpson, J. Neurosci. 20:RC63 (2000) 1-6]. Specifically, these studies implicated oscillatory activity in the alpha frequency-band (8-14 Hz) as an anticipatory mechanism for suppressing distracter visual stimulation. The current study extends these findings by showing that this alpha-suppressive effect is also invoked by cross-modal cues. Auditory symbolic cues were used in an intermodal attention task, to direct subjects' attention to a subsequent task in either the visual or auditory modality. Cueing attention to the auditory features of the imminent task stimuli resulted in significantly higher parieto occipital alpha amplitude in the period preceding onset of this stimulus than when attention was cued to the visual features. Topographic mapping suggests that this effect is generated in regions of the inferior parietal cortex, areas that have been repeatedly implicated in the engagement and maintenance of visual attention. Taken together, the results of this series of studies suggest that these parietal regions are capable of integrating sensory cues from multiple sensory modalities in order to program the subsequent deployment of visual attention. PMID- 11489618 TI - Spatial coordinates of human auditory working memory. AB - The accuracy of localizing remembered sound sources was investigated by employing a delayed-response task, where a small light spot, projected onto a screen by a laser diode attached to the head, had to be spatially aligned with either actual or remembered stimulus positions. Systematic errors indicated overestimation of the eccentricity of remembered targets compared to direct stimulus localization. This overestimation increased with prolonged response delay, suggesting that the coordinates of memorized space are distorted with respect to perceived actual sound location and that this distortion increases as a function of time. PMID- 11489619 TI - Cortical activity associated with vocalization and reading proper. AB - To determine whether motor and premotor areas are involved in silent reading, we report a positron emission tomography study on reading aloud and covert reading. The anterior insula, primary sensorimotor cortex and supplementary motor area were activated separately in reading aloud compared with covert reading, but they were not activated in covert reading compared with fixation control, nor in a conjunction involving reading aloud and covert reading, suggesting the role of articulation or vocalization. PMID- 11489620 TI - Cortical responses to object-motion and visually-induced self-motion perception. AB - We investigated the spatiotemporal cortical dynamics during the perception of object-motion and visually-induced self-motion perception in six normal subjects, using a 143-channel neuromagnetometer. Object-motion specific tasks evoked early transient activity over the right temporooccipital cortex, while self-motion perception, or vection, additionally was followed by sustained bilateral activity in the temporoparietal area. The specific signal distributions suggest to represent the different perceptual modes of object-motion and self-motion sensation. PMID- 11489621 TI - Effects of sequential and temporal probability of deviant occurrence on mismatch negativity. AB - The mismatch negativity (MMN) increases in amplitude as the probability of deviant occurrence decreases. It is unclear whether the determining variable is sequential probability (i.e. the probability of a deviant within a number of standards) or temporal probability (i.e. the probability of a deviant within a period of time). Eight subjects heard a train of frequently occurring 1000 Hz standard tones. The probability of a 1100 Hz pitch deviant was manipulated. In one condition the stimulus-onset-asynchrony (SOA) was 150 ms, with temporal probability of deviant occurrence being either 1/9.00, 1/4.50, 1/2.25, or 1/1.125 s (sequential probability being 1/60, 1/30, 1/15, or 1(deviant)/7.5(standards), respectively). In another condition the SOA was 600 ms, with temporal probability being either 1/9.00, 1/4.50, or 1/2.25 s (sequential probability being 1/15, 1/7.5, or 1/3.75, respectively). In a final condition, the SOA was 2400 ms with temporal probability being 1/9.00 s (sequential probability 1/3.75). Both sequential and temporal probabilities had a marked effect on the MMN. When a deviant occurred every 2.25, 4.50, or 9.00 s, the MMN increased as temporal probability decreased. When a deviant occurred once every 7.5 or 15 standards, the MMN was larger for lower sequential probability, but the effect was not significant. Nevertheless, when temporal probability was held constant at 1/9.00 s, the MMN increased as sequential probability decreased. At rapid rates of stimulus presentation, the MMN was largest. However, it was attenuated when the probability of deviant occurrence was very high perhaps due to the refractoriness of its generator. At the slowest rate, the MMN was diminished perhaps due to memory decay for the standard stimuli. PMID- 11489622 TI - Clathrin-binding proteins: got a motif? Join the network! AB - Clathrin plays a key function in membrane and protein trafficking through the endocytic and late secretory pathways. Its role as a molecular scaffold that drives formation of transport vesicles requires binding to a number of proteins with distinct functional and structural properties. Recent studies have revealed that most of these proteins interact with clathrin through surprisingly simple, linear arrangements of acidic and hydrophobic amino acid residues. This article discusses the different types of clathrin-binding proteins and motifs as well as the physiological significance of these proteins in clathrin-dependent events. PMID- 11489623 TI - The GGAs strike again: cargo capture! PMID- 11489624 TI - A tale of two cultures. PMID- 11489625 TI - Apoptosis: don't get mad, get even! PMID- 11489626 TI - Turning the current up on AMPA receptor trafficking. PMID- 11489627 TI - Histones hold on to RCC1. PMID- 11489628 TI - Zyg-1: elegans personified. PMID- 11489629 TI - First EMBO Young Investigators announced. PMID- 11489630 TI - Job losses in drug development. PMID- 11489632 TI - Rotating chromosomes with lasers. PMID- 11489631 TI - New Royal Society fellows. PMID- 11489635 TI - More noise in the Public Library of Science. PMID- 11489636 TI - Chips help diagnosis of childhood cancers. PMID- 11489637 TI - Synaptic vesicles: is kissing a matter of competence? AB - The "kiss-and-run" model of exocytosis and endocytosis predicts that synaptic vesicles can undergo fast and efficient recycling, after fusion with the plasmalemma, without intermixing of membranes. Evidence is mounting from several new experimental approaches that kiss-and-run occurs at synapses. Distinct vesicle pools, which initially were identified in morphological terms, are now being characterized in biochemical and functional terms. In addition, at least two functional recycling pathways, operating on different time scales (from milliseconds to tens of seconds), have been shown to coexist in the same synaptic system, and the two pathways appear to be differentially regulated. Taken together, these data suggest that kiss-and-run operates in parallel with the classical, coated-vesicle recycling. Here, we review recent evidence for kiss-and run recycling and discuss whether it is a distinct process, dependent on the molecular organization of the fusing vesicle. We propose that vesicles undergo a process of "competence maturation". According to this view, the specific molecular make-up of the vesicles, their location and their interactions with nerve terminal proteins might determine not only the differential availability of the vesicles for fusion and neurotransmitter release but also the recycling path that they will follow. PMID- 11489638 TI - Probing molecular processes in live cells by quantitative multidimensional microscopy. AB - Modern light microscopy has become a most powerful analytical tool for studying molecular processes in live cells. Recent advances in sample preparation, microscope design and image processing allow the generation of "multidimensional" data, simultaneously reporting the three-dimensional distribution and concentrations of several different molecules within cells and tissues at multiple time points with sub-micron spatial resolution and sub-second temporal resolution. Thus, molecular interactions and processes that were approached by biochemical analyses in vitro can now be directly monitored in live cells. Here, we address different aspects of multidimensional microscopy and, in particular, image quantification and the characterization of molecular dynamics, as applied to the study of cell adhesion. PMID- 11489639 TI - Mechanisms that regulate mechanosensory hair cell differentiation. AB - Hair cells of the vertebrate inner ear are mechanosensors that detect sound, gravity and acceleration. They have a specialized cytoskeleton optimized for the transmission of mechanical force. Hair cell defects are a major cause of deafness. The cloning of disease genes and studies of model organisms have provided insights into the mechanisms that regulate the differentiation of hair cells and their cytoskeleton. The studies have also provided new insights into the function of receptors such as integrins and protocadherins, and cytoplasmic proteins such as Rho-type GTPases and unconventional myosins, in organizing the actin cytoskeleton. PMID- 11489640 TI - Signaling pathways in apoptosis as potential targets for cancer therapy. AB - Genetic instability contributes to the origin of cancer as well as to the ability of cancer cells to become resistant to various therapies. Because of this, cytotoxic rather than cytostatic therapies might be most effective against this disease. Many oncogenes and tumor suppressors mediate their effects by interfering with or inducing apoptotic signaling. Thus, apoptotic pathways might be significantly altered in cancer cells relative to untransformed cells, and these differences might present a therapeutic window that can be exploited for development of cancer drugs. PMID- 11489641 TI - Gunter Blobel--still passionate after all these years. AB - What one word best describes Gunter Blobel? ANSWER: passion. Almost all of the nearly 200 investigators who have worked in the Blobel laboratory in the past quarter century would describe him as highly enthusiastic, intense and, above all, passionate. Whether it was in the early days of the signal hypothesis, the initial characterization of nuclear pores and lamina or his more recent foray into rebuilding the historic German City of Dresden, Gunter attacks every project with unbridled passion, intensity, boundless energy and determination. PMID- 11489642 TI - Aortic graft infections: replacement with autogenous vein. AB - The purpose of this review article is to summarize our published experience with the use of the superficial femoral-popliteal vein (SFPV) to replace infected aortic prostheses. The SFPV has proven to be resistant to infections of all types and has shown no signs of degeneration over the long term. Since SFPV bypass and prosthetic graft excision are performed as a single stage, operative times are extensive. Therefore, it may not be appropriate for the sickest patients with severe medical comorbidities. Nevertheless, the operation has been associated with gratifyingly low mortality and amputation rates that are far better than published rates associated with graft excision and extra-anatomic bypass. It is particularly suited to patients with complex aortofemoral graft reconstructions who cannot undergo extra-anatomic revascularization for technical reasons. The venous sequella of SFPV harvest are minimal. These data and those from three other centers support the conclusion that graft excision and replacement with SFPV is an excellent alternative for treatment of aortic graft infections. PMID- 11489643 TI - Thromboprophylaxis in an academic medical center. AB - INTRODUCTION: Questions regarding which patients require prophylaxis for thromboembolism, what methods should be used and the appropriate duration of treatment remain unanswered. METHODS AND MATERIALS: A retrospective review from a single academic medical center was undertaken to evaluate prophylactic strategies. Multiple sources of data were used to identify patients who were prophylaxed and those who developed deep vein thrombosis or pulmonary embolism. These data were analyzed to determine factors associated with successful prophylaxis including age, type of prophylaxis and admitting services. RESULTS: A total of 22,030 patients were admitted of whom 7520 (36%) received prophylaxis and there were 523 thromboembolic events. Pneumatic compression devices and antiembolic stockings had the lowest incidence of failure, 2.2% and 3.2% respectively. There were significant differences in the rates of prophylaxis used by the five admitting services, being highest in surgery (40.8%) and lowest in gynecology (11.4%). However, these groups had the lowest incidence of venous thromboembolism (4.3%, 2.3%). Both of these groups used pneumatic compression as the method of choice (64.3% and 65.2%). CONCLUSIONS: Thromboembolism prophylaxis reduces the incidence of DVT and PE, however, our study demonstrates the variable effectiveness of each method in different types of patients. Our data suggest that patient risk of DVT should be individually assessed and an appropriate method of prophylaxis should be applied when warranted. PMID- 11489644 TI - Growing the vascular surgical practice: venous disorders. AB - For the 48th meeting of the International Society for Cardiovascular Surgery, North American Chapter in June 2000, we were asked to organize a presentation with the title displayed above. We welcomed this opportunity because the phrase 'growing the practice' literally means giving greater service to underserved patients with vascular disorders. Underserved individuals with vascular disorders include those with nonatherosclerotic arterial disorders, patients with lymphedema, and individuals with a variety of venous disorders including venous insufficiency. The majority of these conditions are not surgical, but venous insufficiency commands attention because its treatments are interventional. Therefore, this presentation will discuss prevalence of venous insufficiency, a unifying concept of its pathophysiology, how treatment may be selected, what the new technology provides, and then a description of coding issues and whether or not interventions are actually worthwhile from the point of view of the patient. PMID- 11489645 TI - Surgical outcome of infectious aneurysm of the abdominal aorta with or without SIRS. AB - The surgical outcome of infectious abdominal aortic aneurysms was evaluated based on the preoperative presence or absence of systemic inflammatory response syndrome (SIRS). Nine patients were divided into two groups according to the criteria for SIRS such as body temperature, heart rate, respiratory rate, and white blood cell count. In the group with SIRS, rupture and impending rupture of aneurysms occurred in three of the four patients (75%). All aneurysms were resected with a small part as a remnant; two in situ and two extraanatomic reconstructions were performed. Three patients died after surgery: one after in situ (cerebral infarction) and two after extraanatomic reconstruction (sepsis and multiple organ failure). In the group without SIRS, closed en bloc resection in two patients and resection of the aneurysm with a small part as a remnant in three patients were performed. In situ reconstruction in all patients and omentum wrapping in two patients were performed. One of the five patients died of massive hematemesis 70 days after surgery. The overall mortality rate was 75% in the group with SIRS versus 20% in the group without SIRS. The surgical outcome of infectious abdominal aortic aneurysm depends upon the severity of underlying infection. A possibility exists that SIRS is a useful indicator for predicting the surgical outcome of patients. PMID- 11489646 TI - The cost-effectiveness of exclusion arteriography in extremity trauma. AB - The purpose of this study was to apply decision analysis to an established practice in vascular trauma diagnosis. While exclusion arteriography has resulted in an increase in positive surgical explorations, no formal analysis that determined either the cost-effectiveness of exclusion arteriography or the cost effectiveness ratio has been reported in the literature. We created a decision model that compared exclusion arteriography and surgical exploration, the standard used prior to the development of extremity arteriography. The decision model used predominantly literature derived estimates for the prevalence of arterial injuries and the accuracy, complications, outcomes and costs of both arteriography and exploration. Exclusion arteriography is cost-effective. This finding is robust to changes in the major model variables. Compared to surgical exploration, exclusion arteriography is a superior strategy by dominance (more effective and costs less). Therefore, a cost-effectiveness ratio cannot be calculated. Under the base case assumption of 28% prevalence of arterial injury requiring operation, exclusion arteriography saves about $2000 and adds 0.3 quality adjusted life years (QALY) for each patient. Decision analysis can be successfully applied to problems in vascular trauma diagnosis. PMID- 11489647 TI - Vascular ultrasound surveillance after endovascular intervention for occlusive iliac artery disease. AB - This paper describes vascular ultrasound surveillance after endovascular intervention for occlusive iliac artery disease. There were 105 patients who had 198 procedures in 155 limbs, consisting of 110 balloon dilatations and 88 stentings. The patients were referred to the vascular diagnostic service by several surgeons. All procedures had been considered to be initially technically successful. Colour-Doppler duplex ultrasound studies were performed shortly before and at serial intervals after operation to determine patency rates. Univariate life table analysis showed 69% primary and 96% assisted primary patency at four years. Primary patency rates at four years were significantly worse for stentings (60%) compared to balloon dilatations (71%) (P<0.05). Maximum peak systolic velocities (PSV) were recorded from the treated arteries. Receiver operating characteristics curves showed that PSV >300 cm/s was most accurate for predicting technical failure. Haemodynamic success rates at four years were 72% for PSV >300 cm/s. Results for procedures that were initially successful indicate that long-term primary patency rates for iliac endovascular intervention are acceptable and that assisted primary patency rates are excellent. PMID- 11489648 TI - A prospective evaluation of the risk for venous leg thrombosis associated with prolonged air travel: a pilot study. PMID- 11489649 TI - Carotid endarterectomy with bovine patch angioplasty: a preliminary report. AB - Carotid endarterectomy with patch angioplasty is a durable procedure for prevention of recurrent neurological symptoms and stroke. However, no definitive study has demonstrated a clear benefit of one class of the patch material over another. The aim of this study was to evaluate the clinical outcome of carotid endarterectomy with bovine pericardium patch in comparison with autologous vein patch. One hundred and twenty-two carotid endarterectomies were performed using patch closure of the arteriotomy with bovine pericardium (61 cases) and autologous vein (61 cases) between September 1995 and June 1999. Though this is not a double-blind type randomized comparative study, effort was made to achieve a 1:1 ratio in sequence with a few exceptions such as non-available veins at time of surgery or for future use. In bovine pericardium patch closure group, the mean total operating time was significantly shorter than autologous vein closure group (P<0.01), but the mean carotid clamping time was similar in both groups, regardless of the use of shunt. The incidence of postoperative local complications including groin wound was less in bovine pericardium patch closure group. The patients were followed with duplex scans in one month post-CEA and 6 months interval thereafter. During the follow-up period, three patients developed non-critical stenosis (two in bovine and one in vein patch), and one aneurysmal dilation occurred in vein patch group. The incidence of restenosis was similar in both groups. Although this is a preliminary report, it is concluded that the results of carotid angioplasty using bovine pericardium compare favorably with autologous vein. PMID- 11489650 TI - Six hundred consecutive carotid endarterectomies with temporary shunt and vein patch angioplasty: early and long-term results. AB - PURPOSE: This retrospective study evaluates the long-term clinical outcome and the survival of 600 consecutive carotid endarterectomies performed with a temporary shunt. All arteriotomies were closed by vein patch angioplasty. MATERIALS AND METHODS: Between November 1989 and November 1998, 600 isolated carotid endarterectomies (CEA) were performed in 540 patients by a uniform surgical technique at the University Clinic of Mont-Godinne. An intraluminal shunt and patch closure were systematically used. The mean age was 68 yr (ranging from 41 to 91 yr), 400 patients were men. The risk factors included hypertension in 73%, smoking history in 60%, coronary artery disease in 51% and hyperlipidemia in 35%. The indications were asymptomatic stenosis in 47%, transient ischemic attack in 40%, vertebrobasilar symptoms in 7% and stroke in 6%. EARLY RESULTS: The combined 30-day stroke and death rate was 0.9%. There were four deaths. The stroke and TIAs rates were 0.2% and 1.5% respectively. The incidence of early carotid occlusion was 0.5%. Cranial or cervical nerve dysfunction was identified in 6.3%. LATE RESULTS: The median follow-up was 49 months with a range of 2-124 months. Cumulative survival rates at 5 and 10 yr were 92+/-1% and 89+/-2% respectively. Thirty-two patients died during long-term follow-up; the death was stroke-related in only three patients. CONCLUSION: Carotid endarterectomy using an intraluminal shunt and vein patch closure is a safe and effective procedure associated with low morbidity and mortality rates at short and long-term follow up. PMID- 11489651 TI - Surveillance after endoluminal repair of abdominal aortic aneurysms. PMID- 11489652 TI - Combined upper and lower extremity deep venous thrombosis. AB - In order to elucidate the natural history of upper extremity deep venous thrombosis (UEDVT), we compared the morbidity and mortality of patients with UEDVT and that of patients with both UEDVT and lower extremity deep venous thrombosis (LEDVT). Between 1993 and 1996, 21 patients presented to our institution with both LEDVT and UEDVT (Group 1). During the same time period, 144 patients were diagnosed with UEDVT alone (Group 2). The diagnosis was confirmed by duplex scanning in all patients. In Group 1, there were 14 females (67%) and 7 males (23%) with ages ranging from 25 to 97 yr old [mean 73 yr old +/-17 yr (SD)]. In Group 2, there were 84 females (58%) and 60 males (42%) with ages ranging from 9 to 101 yr old [mean 67 yr old +/-17 yr (SD)]. Differences in age and sex between the two groups were not statistically significant. In Group 1, systemic anticoagulation was implemented in 17 patients (81%). Two patients (9.5%) required placement of a SVC and IVC filters due to contraindication to anticoagulation. One patient did not receive anticoagulation, and one patient was only started on aspirin. In Group 2, treatment consisted of systemic anticoagulation in 94 patients (65%). The remainder of the patients were treated with aspirin in three patients (2%) or no anticoagulation in 31 patients (19%). Sixteen patients (11%) underwent placement of a SVC filter either due to failure of anticoagulation to prevent pulmonary embolism (two patients) or contraindication to anticoagulation (14 patients). Pulmonary emboli were documented by ventilation/perfusion lung scan in two patients (9.5%) in Group 1 and in 16 patients (11%) in Group 2. In the first group, 8 of the 21 patients (38%) were dead within 1 month of the diagnosis of UEDVT, and 11 of 21 patients (52%) were dead within 2 months of the diagnosis of UEDVT. In the second group, 20 of 144 patients (14%) were dead within 1 month of the diagnosis of UEDVT and 38 of 144 patients (26%) were dead within 2 months of diagnosis (P<0.02). Our data suggest that patients with both UEDVT and LEDVT have a higher mortality than patients with UEDVT alone. As the risk for pulmonary embolism is similar in both groups, we speculate that the severity of medical illness in patients with both UEDVT and LEDVT may contribute to the higher mortality. This is the first study to examine the mortality of this group of patients. PMID- 11489653 TI - Oral anticoagulation therapy during pregnancy in patients with mechanical mitral valves: a prospective study. AB - Reports on phenindione toxicity have limited its use as an oral anticoagulant. Our aim was to evaluate its risks in pregnant women. Thirty-one pregnancies in 29 women with mitral (+/-aortic) St. Jude mechanical valves were followed-up prospectively. Eighteen patients received phenindione. Eleven patients (37.9%) received in addition to phenindione 225 mg dipyridamole, which was given in three doses. The target INR was 2.5-3.5 in the former and 2-2.5 in the latter treatment. A fortnight before delivery, intravenous heparinotherapy was substituted. There were no maternal complications, apart from a single postpartum hemorrhage (3.2%). After the deliveries the results were: 26 mature babies (83.9%), 3 premature babies (9.7%) and 2 cases of stillbirth (6.4%). Outcome was dose related; being 57.2+/-20.9 mg/day for mature babies and 82.5+/-11.2 mg/day for prematures and stillbirths (P=0.016). Phenindione provided safe and effective anticoagulation during pregnancy. A larger study is necessary to confirm the relationship between the dosage and outcome. PMID- 11489654 TI - Impact of myocardial protection during coronary bypass surgery on patient outcome. AB - We have recently shown that continuous coronary perfusion with warm blood enriched with the ultra-short acting beta-blocker Esmolol (ES) improves functional and structural myocardial protection during coronary artery surgery as compared with conventional cardioplegia (CP). The purpose of the present study was to compare both myocardial protection techniques in terms of patient outcome. We retrospectively analyzed the charts of 150 consecutive patients subjected to coronary artery surgery using the ES-technique; 150 patients matched for age, gender, preoperative left ventricular function, history of renal failure, and history of neurological symptoms undergoing surgery with conventional CP during the same time period served as control group. There were no significant differences between both groups with respect to perioperative myocardial infarction rate, need for positive inotropic medication, need for mechanical circulatory support, duration of mechanical ventilation, duration of intensive care unit stay, time of mobilization, postoperative renal failure, cardiac arrhythmias, neurological symptoms, infections or in-hospital mortality. ES patients were less frequently readmitted to the intensive care unit (ES: 3/150; 2.2% [95% confidence interval: 0-4.2%] vs. CP: 13/150; 8.7% [4.2-13.2%]; P=0.010) and total hospital stay was shorter (ES: 12.3+/-4.8 days [95% CI: 11.5-13.0] vs CP: 13.5+/-3.8 [12.9-14.1] days; P=0.0013), thus saving 159 patient days on the normal ward. Procedural costs were less for the ES-technique (US$ 60 per patient) as compared to the cardioplegia technique (US$ 120 per patient). These data suggest that myocardial protection using the ES-technique does not improve clinical outcome in patients subjected to routine coronary artery surgery, but may save costs. PMID- 11489655 TI - Coronary artery bypass grafting in octogenarians. AB - Preoperative profiles, postoperative complications, and the early and late results in 32 patients 80 yrs. of age and older (elderly group) who underwent coronary artery bypass grafting were compared with those in patients under 80 yrs. of age (control group). In the elderly group, the prevalence of patients with preoperative creatinine clearance (Ccr.) <50 l/day (34.4%), unstable angina pectoris (78.1%) and left main trunk disease (40.1%) was significantly higher than those in the control group. The incidences of arrhythmia and intensive care unit(ICU) syndrome were also significantly higher in the elderly group than in the control group, however, there was no death due to these complications. In the elderly group, one patient (3.1%) died in the hospital due to low cardiac output syndrome (LOS), while three patients (2.4%) of the control group died in the hospital. As for the long-term results, the 5-yr. survival rates for the elderly group and the control group were 82.6% and 85.2%, respectively, and the effectiveness of surgery was remarkable, with improved postoperative activity in 96.9% of the elderly group. These findings indicate that although the elderly patients have higher risks by undergoing surgery and have a disadvantage in the rate of postoperative complications, the postoperative improvement in activity and survival rate can be similar to those in the younger patients. PMID- 11489656 TI - Expression of vascular endothelial growth factor in aortocoronary saphenous vein bypass grafts. AB - Neovascularisation is a prominent feature of long-term aortocoronary saphenous vein bypass grafts but mechanisms involved in the formation of neovessels have not been previously studied. Vascular Endothelial Growth Factor (VEGF) is an important angiogenic factor that induces migration and proliferation of endothelial cells, enhances permeability and modulates thrombogenecity. This study investigated the expression of VEGF in aortocoronary saphenous vein bypass grafts. Aortocoronary saphenous vein bypass grafts with angiographic luminal stenosis of >75% were explanted from 14 patients at redo coronary artery bypass grafting. The grafts demonstrated two distinct forms of graft occlusion: four out of the 14 graft occlusions (29%) resulted from severe hyperplastic transformation of the intima complicated by thrombi attached to the degenerating liminal endothelium; the remaining graft occlusions (71%) were due to the development of atherosclerotic lesions associated with mural thrombosis. Hiperplastically altered intimal segments were practically free of neovascularisation while atherosclerotic-like lesions contained neovessels irregularly distributed throughout. Intimal neovessels were located exclusively in microzones enriched with VEGF-expressing cells and, furthermore, neovascular endothelial cells themselves also displayed VEGF immunopositivity. Double-immunostaining revealed that in areas of neovascularisation, the vast majority macrophages (CD68+) expressed VEGF. Some CD68+ foam cells that surrounded branches of neovascularisation were also VEGF-positive. These findings suggest that VEGF expressed by neovascular endothelial cells and by macrophages may act as a local regulator of endothelial cells functions and may induce intimal neovascularisation in aortocoronary saphenous vein bypass grafts affected by atherosclerosis. PMID- 11489657 TI - A simple method to determine anastomotic quality of coronary artery bypass grafting in the operating room. AB - Anastomotic quality of coronary artery bypass grafting is directly associated with peri-operative and long-term clinical results. In this study, we investigated a cut-off value for intra-operative flow measurement. This value could be of use to a surgeon in determining the anastomotic quality of grafts. Intra-operative transit-time flow variables (mean flow, pulsatility index, % efficiency, fast Fourier transformation (FFT) of the flow curve) and the 2-week post-operative angiographic findings were examined in 66 coronary artery bypass grafts, including 33 internal thoracic arteries. There were significant differences between patent and non patent grafts in all of the intra-operative flow parameters. Only the FFT ratio, the ratio of powers of the fundamental frequency and its first harmonic, could be utilized as a cut-off value to distinguish patent from non patent grafts. All stenotic or occluded grafts showed an intra-operative FFT ratio of <1.0, while all patent grafts yielded a ratio of >1.0. Based upon these results, we concluded that power spectral analysis of flow measurement might be useful for intra-operative differential diagnosis of the anastomotic quality in coronary artery bypass grafting. PMID- 11489658 TI - The impact of hypertension on the operative and early postoperative outcome of aortic valve surgery. AB - Hypertension is a known risk factor in heart disease. It can lead to pressure overload and hypertrophy of the left ventricle. The aim of this study is to examine the effect of hypertension on the operative and early postoperative outcome after aortic valve surgery using the retrograde cardioplegia. All the data of all the patients who had aortic valve surgery in our department were retrospectively examined during the period from January 1994 until April 1996 and received retrograde blood cardioplegia. 397 patients were included in this study. 213 of them had arterial hypertension, as preoperatively diagnosed by the referring cardiologist. There were 163 females and 234 males. 142 were above 70 yr of age. 22 patients had an ejection fraction (EF) < or =0.4 and in 168 patients the LVEDP was >15 mmHg. Hypertension alone proved to be no risk factor. Decreased EF in hypertensive patients leads to an increase in the occurrence of prolonged ICU-stay, low cardiac output and neurological complications. Hypertension alone does not increase the risk of operative and early postoperative aortic valve surgery. PMID- 11489659 TI - Emergency coronary artery bypass grafting: does excessive preoperative anticoagulation increase bleeding complications and transfusion requirements? AB - Patients requiring urgent surgical revascularization due to unstable coronary artery disease are usually pretreated with multiple antithrombotic drugs. The perioperative risks of this type of treatment were investigated in 123 patients who underwent emergency coronary artery bypass grafting (CABG) at our institution.Eighty-two patients (group A) received heparin and acetylsalicylic acid solely and 41 patients (group B) received additionally ADP-receptor antagonists (82.9%), glycoprotein IIb/IIIa inhibitors (12.2%) or thrombolysis (14.6%) preoperatively. Both groups were similar regarding demographic data and overall clinical status. Preoperative coagulation parameters and intraoperative characteristics were comparable. Blood loss via chest tubes was not significantly different between groups. Transfusion of red blood cells and fresh frozen plasma were slightly, but not significantly increased in group B. Transfusion of pooled platelets was low in general and similar in both groups. Re-exploration rate, medium intensive care unit and hospital stay as well as perioperative mortality were comparable.Excessive antithrombotic pretreatment seems to bear no additional risk in emergency CABG and may be beneficial in this setting. PMID- 11489661 TI - Analytical pharmacology of G protein-coupled receptors by stoichiometric expression of the receptor and G(alpha) protein subunits. AB - The description of a new family of recombinant proteins, which are constructed by the covalent fusion of the cDNA encoding a G protein-coupled receptor with that of a G(alpha) protein subunit, has recently been introduced as an original strategy to explore receptor pharmacology under defined experimental conditions. As such, a controlled 1:1 stoichiometry of receptor and G(alpha) protein expression can be achieved, as well as a forced spatial proximity to each other. Fusion proteins have been revealed as active at the receptor ligand binding level and functional at the G(alpha) protein and effector level. Insights on analytical pharmacological data are discussed for wild-type and mutant receptors interacting with a given G(alpha) protein subunit and different subtypes of either wild-type or mutant G(alpha) proteins activated by a single receptor subtype. A possible alteration of the receptor:G(alpha) protein selectivity may occur due either to the spatial proximity of both protein partners or to a constraint receptor state unable to accommodate to different G(alpha) protein states. Coactivation of endogenous G(alpha) proteins in host cells expressing a fusion protein has also been observed, but depends mainly on the coupling efficiency of the receptor and G(alpha) protein engaged in the fusion process. The ligand's apparent intrinsic activity has been shown to be either enhanced, attenuated, or unmodified when the functional responses of a fusion protein are compared to the coexpression of both fusion protein partners. PMID- 11489663 TI - An apparatus to assay opioid activity in the infused lumen of the intact isolated guinea pig ileum. AB - A modified apparatus is described that provides for the simultaneous bathing of the serosa of an intact piece of isolated guinea pig ileum while allowing infusion of the isolated lumen. The comparative compartmental potency of the opioid agonists morphine, casomorphins, and enkephalins to inhibit electrically driven contractions are described in this system. The rank-order potency for serosally applied opioid agonists was (IC(50) values, nM): [D-Ala(2),N-Me Phe(4),Gly-ol(5)]-enkephalin (DAMGO) (15)>[D-Ala(2),D-Leu(5)]-enkephalin (DADLE) (35)> or =morphine (46)> or =[D-Ala(2)]-met-enkephalinamide (55)>[D-Ala(2)]-beta casomorphin[1--4] amide (122)>beta-casomorphin[1--4] amide (940)>met- and leu enkephalin (>6000). This contrasted to the rank-order potency for the luminally applied opioid agonists: DADLE (63)>DAMGO (135)>[D-Ala(2)]-met enkephalinamide=morphine (4700)>[D-Ala(2)]-beta-casomorphin[1--4] amide (29000). beta-Casomorphin[1--4] amide, leu-enkephalin and met-enkephalin are mostly inactive when applied luminally. Furthermore, the opioid antagonists, casoxin 4 and [D-Ala(2)]-casoxin 4, when infused into the lumen, significantly overcame the inhibitory effect of morphine added to the serosal side. This model provides an assay and screening system to differentiate between the effects of chemical agents applied via the blood stream (serosa) or food side (lumen) on quiescent or electrically driven gut activity of the nervous plexi or receptor systems of the ileum. PMID- 11489662 TI - Nonlinear analysis of partial dopamine agonist effects on cAMP in C6 glioma cells. AB - Most drugs have some efficacy so that improved methods to determine the relative intrinsic efficacy of partial agonists should be of benefit to preclinical and clinical investigators. We examined the effects of partial D(1) or partial D(2) dopamine agonists using a partial agonist interaction model. The dependent variable was the modulation of the dopamine-receptor-mediated cAMP response in C6 glioma cells selectively and stably expressing either D(1) or D(2) recombinant dopamine receptors. The dissociation constant (K(B)) and relative intrinsic efficacy (E(r)) for each partial agonist were calculated using a partial agonist interaction null model in which the effects of fixed concentrations of each partial agonist on the dopamine dose-response curve were evaluated. This model is an extension of the competitive antagonist null model to drugs with efficacy and assumes only that the log-dose--response curve is monotonic. Generally, the partial agonist interaction model fit the data, as well as fits of the independent logistic curves. Furthermore, the partial agonist K(B) values could be shared across partial agonist concentrations without worsening the model fit (by increasing the residual variance). K(B) values were also similar to drug affinities reported in the literature. The model was validated in three ways. First, we assumed a common tissue stimulus parameter (beta) and calculated the E(r) values. This provided a qualitative check on the interaction model results. Second, we calculated new relative efficacy values, E(r)(beta), using the beta estimate. Third, we calculated relative efficacy using relative maxima times midpoint shift ratios (J. Theor. Biol. 198 (1999) 347.). All three methods indicated that the present model yielded reasonable estimates of affinity and relative efficacy for the set of compounds studied. Our results provide a quick and convenient method of quantification of partial agonist efficacy. Special applications and limitations of the model are discussed. In addition, the present results are the first report of the relative intrinsic efficacy values for this set of D(2) ligands. PMID- 11489664 TI - A flowcytometric method for evaluation of acid secretion from isolated rat gastric mucosal cells. AB - Studies on the gastric proton pump are mostly performed on the H(+), K(+)-ATPase enzyme in the microsomal preparation or by aminopyrine accumulation in the gastric parietal cells. H(+), K(+)-ATPase activity is estimated by both spectrophotometric and fluorimetric methods. In the present study, quenching or augmentation in acridine orange (AO) fluorescence was monitored on a flowcytometer in rat gastric mucosal cells. Rat gastric mucosal cells were isolated by the standard pronase--EDTA method. The effect of oleic acid, a proton pump inhibitory was evaluated on gastric parietal cell activity and was compared with its effect on proton transport, H(+), K(+)-ATPase, and p-nitrophenyl phosphatase (p-NPPase) activity in gastric microsomes. In addition, the effect of histamine and carbachol, gastric acid release inducers, was also investigated by flowcytometry in isolated parietal cells. Histamine and carbachol, in a dose dependent manner, stimulated acid release from isolated gastric cells. Oleic acid also dose-dependently inhibited the basal and stimulated acid release from the cells, as well as in all three enzyme preparations associated with gastric proton pump activity. Thus, the results suggest that flowcytometric method might be used to study basal, as well as stimulated, proton pump activity in isolated gastric parietal cells. PMID- 11489665 TI - The applicability of rat and human liver slices to the study of mechanisms of hepatic drug uptake. AB - In the present study we investigated the applicability of the liver slice model to study mechanisms of drug uptake. Four model compounds were investigated that enter hepatocytes via entirely different membrane transport mechanisms. Rhodamine B (RB), which enters hepatocytes by passive diffusion, was homogeneously distributed throughout the rat liver slice (250 microm thickness) within 5 min, indicating that the penetration rate into the slice and the diffusion rate into the cells are rapid. In contrast, lucigenin (LU), which is taken up by hepatocytes through adsorptive endocytosis, was detected in the inner cell layers after 15 min. Digoxin uptake into the slice showed a temperature-dependent component and was stereoselectively inhibited by quinine, which is compatible with the involvement of a carrier-mediated uptake mechanism. The neo-glycoalbumin Lactose(27)-Human Serum Albumin (Lact(27)-HSA) and the negatively charged Succinylated-Human Serum Albumin (Suc-HSA) entered the slices and were taken up temperature-dependently into hepatocytes and endothelial cells, respectively. The liver slice preparation is a valuable tool to investigate the mechanisms of cellular uptake of drugs. Moreover, the precision-cut liver slices offer the unique possibility to study both hepatocyte and endothelial cell function in human and rat liver. PMID- 11489666 TI - Application of in silico approaches to predicting drug--drug interactions. AB - In an environment driven to find the next blockbuster drug, failure years into a project should not be an option. Recent studies have shown that poor absorption, distribution, metabolism, and excretion (ADME), and the related properties of toxicity and pharmacokinetics are responsible for a large proportion of failures. One way to understand and potentially predict molecules likely to be successful in humans as drugs from an ADME point of view is to use simulations. Such simulations may include simple rule-based approaches, structure--activity relationships, three-dimensional quantitative structure--activity relationships (3D-QSAR), and pharmacophores. All of these represent useful tools in understanding metabolism by the cytochromes P450, predicting drug--drug interactions (DDIs), and other pharmacokinetic parameters. The present paper briefly reviews the application of some computational tools applied to predicting DDIs and will provide the reader with an idea of their utility. PMID- 11489667 TI - Evaluation of the BBMEC model for screening the CNS permeability of drugs. AB - Combinatorial synthesis and high-throughput pharmacology screening have greatly increased compound throughput in modern drug-discovery programs. For CNS drugs, it is also important to determine permeability to the blood--brain barrier. Yet, given the increased pace of discovery, it difficult to conduct this screen in a timely fashion. In this presentation, we describe several improvements to an existing CNS permeability screen, the bovine brain microvessel endothelial cell (BBMEC) model. By implementation of these incremental process improvements, we have achieved a robust, facile screen for determination of CNS permeability of multiple compounds. PMID- 11489670 TI - Proceedings of the 5th International Symposium on the Treatment of Parkinson's Disease. September 30-October 1, 2000. Kobe, Japan. PMID- 11489668 TI - Problems associated with in vitro assessment of drug inhibition of CYP3A4 and other P-450 enzymes and its impact on drug discovery. AB - The cytochromes P-450 recognize and metabolize a broad range of structurally diverse therapeutic agents. As a consequence, many clinically relevant drug--drug interactions (DDI) are associated with inhibition and/or induction of a specific P-450 enzymes (in particular human cytochrome P-450 3A4, CYP3A4). In addition to inhibition and induction, CYP-mediated drug metabolism may be enhanced upon coincubation with certain compounds. Moreover, some of these enzyme-based interactions appear to be substrate specific. In this presentation, several issues associated with the generation of accurate DDI information will be discussed. PMID- 11489671 TI - Anatomy and function of dopamine receptors: understanding the pathophysiology of fluctuations in Parkinson's disease. AB - The principal dopamine (DA) receptors mediating the antiparkinson effects of levodopa are D1 and D2, which are anatomically and functionally segregated. We hypothesize that DA receptor-mediated effects are critical for the development of treatment-related fluctuations in Parkinson's disease (PD). We suggest that two sequential processes occur to permit the emergence of the so-called short duration response and dyskinesias: (1) functional uncoupling of D1 and D2 receptor-mediated effects with shift to the left of the dose-response curve; and, (2) sensitization of the D1-mediated striatal output due to repetitive, primarily D1 receptor stimulation by DA. These mechanisms result in excessive, pathological inhibition of basal ganglia output neurons in the GPi producing dyskinesias and the short duration response. PMID- 11489672 TI - Molecular neurotoxicological models of Parkinsonism: focus on genetic manipulation of mice. AB - Parkinson's disease is a neurodegenerative disorder that affects mainly the nigrostriatal dopaminergic system in humans. Several propositions have been put forward to explain the cellular and molecular pathobiology of this syndrome. Initial attempts were made through the use of various agents to manipulate the deleterious effects of toxins that destroy dopaminergic cells both in vitro and in vivo. These studies led to the idea that oxidative stress is an important factor in killing these cells. More recent attempts have made use of genetically modified mice to eliminate or over-express genes of interest. These experiments have suggested that the destruction of dopaminergic cells might be the result of the convergence of dependent and independent molecular pathways and that trigger cellular events might lead to the demise of these dopaminergic cells. PMID- 11489673 TI - Functional brain networks in Parkinson's disease. AB - With the advent of new methods of network analysis, we have utilized metabolic data acquired through positron emission tomography (PET) to identify disease related patterns of functional pathology in the movement disorders. In Parkinson's disease (PD), we have used [(18)F]-fluorodeoxyglucose (FDG)/PET to identify a disease-related regional metabolic covariance pattern characterized by lentiform and thalamic hypermetabolism associated with regional metabolic decrements in the lateral premotor cortex, the supplementary motor area, the dorsolateral prefrontal cortex, and the parieto-occipital association regions. The expression of this network is modulated in a predictable fashion by levodopa therapy and by stereotaxic interventions for PD.We have extended this network analytical approach from studies of glucose metabolism in the resting state to dynamic studies of brain activation during motor performance. These PET studies utilized [(15)O]-water (H(2) (15)O) to measure cerebral blood flow activation responses during the execution of simple and complex motor tasks. In addition to the modulation of abnormal resting metabolic networks, effective PD therapy can enhance brain activation responses during motor execution, with specific regional associations with improvements in timing and spatial accuracy.This approach is also useful in identifying specific brain networks mediating the learning of sequential information. We have found that the normal relationship between brain networks and learning performance are altered in the earliest stages of PD with a functional shift from striatal to cortical processing. Brain activation PET studies during therapeutic interventions for PD demonstrate how normal brain behavior relationships can be restored with successful therapy. Thus, functional brain imaging with network analysis can provide insights into the mechanistic basis of basal ganglia disorders and their treatment. PMID- 11489674 TI - Levodopa prolongs life expectancy and is non-toxic to substantia nigra. AB - The primary objective of the study was to determine the effect of levodopa (LD) on human substantia nigra. The study included patients seen at the Movement Disorder Clinic, Saskatoon over a 32 year period. The evidence provided is based on epidemiological observations of 934 consecutive Parkinson syndrome (PS) patients assessed during 22 years and detailed studies of six patients including two autopsies. Life expectancy increased significantly with LD therapy. The prolonged survival was evident when the patients were treated during early stage of the illness. One parkinsonian patient with substantia nigra (SN) pathology who was extensively studied for 30 years, revealed significant slowing of the disease progression while on LD. Three essential tremor patients who received 24 kg (26 years), 22 kg (21.5 years), and 8.5 kg (12.5 years) LD respectively, had no evidence of PS and one autopsy revealed normal SN. Two dopa-responsive dystonia patients who received LD 3 kg (11 years) and 17 kg (29 years) each had no evidence of PS and one autopsy revealed normal number of SN neurons. These observations indicate that LD is not toxic to human SN and are consistent with salutary effect of the drug on the SN in Parkinson's disease. PMID- 11489675 TI - Motor fluctuations and dyskinesia in Parkinson's disease. AB - Motor fluctuations and dyskinesia are common complications of long-term levodopa therapy. The neural and molecular mechanisms underlying their development are partially understood. A variety of clinical strategies may reduce the unpredictability of motor fluctuations and reduce their impact. Prevention of these complications remains an elusive goal. PMID- 11489676 TI - Dystonia and parkinsonism. AB - Parkinsonism and dystonia may coexist in a number of neurodegenerative, genetic, toxic, and metabolic disorders and as a result of structural lesions in the basal ganglia. Parkinson's disease (PD) and the 'Parkinson-plus' syndromes (PPS) account for the majority of patients with the parkinsonism-dystonia combination. Dystonia, particularly when it involves the foot, may be the presenting sign of PD or PPS and these disorders should be suspected when adults present with isolated foot dystonia. Young age, female gender, and long disease duration are risk factors for PD-related dystonia, but dystonia in patients with PD is usually related to levodopa therapy. The mechanism of dystonia in PD is not well understood and the management is often challenging because levodopa and other dopaminergic agents may either improve or worsen dystonia. Other therapeutic strategies include oral medications (baclofen, anticholinergics and benzodiazepines), local injections of botulinum toxin, intrathecal baclofen, and surgical lesions or high frequency stimulation of the thalamus, globus pallidus, or subthalamus. PMID- 11489677 TI - Approaching disturbed sleep in late Parkinson's Disease: first step toward a proposal for a revised UPDRS. AB - A patient in stage 3-4 of the Unified Parkinson's Disease Rating Scale (UPDRS), or in stage 4-5 of Hoehn and Yahr staging scale, or a patient with 0-50% activities of daily living scale of Schwab and England is considered a Late Parkinson's Disease (LPD) patient. The prevalence of disturbed sleep in Parkinson's Disease (PD) was found to vary according to an objective rating, from 60 to 98%. The factors predicting the quality of life in PD patients are: depression, sleep disturbances and dependence. The present article proposes the insertion of the following items as a chapter in a revised UPDRS based on updated knowledge in sleep arousal disturbances in PD. V. SLEEP-AROUSAL DISTURBANCES: Sleep disturbances 43. Light fragment sleep (LFS) 44. Sleep-related breathing disorders (SRBD) 45. Restless legs-periodic leg movements during sleep (RLS-PLM) 46. REM behavioral disorders (RBD) 47. Sleep-related hallucinations (SRH) 48. Sleep-related psychotic behavior (SRPB) Arousal disturbances 49. Sleep attacks (SA) 50. Excessive daytime sleepiness (EDS). Approaching the treatment of disturbed sleep in LPD means postponement of the institutionalization of the LPD patient, allowing the spouse or the caregiver a quiet nights sleep. This approach consists of three steps, each one of major importance. (1) Correct diagnosis based on detailed anamnesis of the patient, of the spouse or of the caregiver; a one week recording on a symptom diary (log) by the patient or the caregiver; excluding co morbidities. Then choosing the most appropriate sleep test, if necessary: polysomnography (PSG), multiple sleep latency test (MSLT), multiple wake latency test (MWLT), actigraphy or video-PSG. This first step allows the diagnosis of one of the above mentioned sleep-arousal disturbances. (2) The non specific therapeutic approach consists of: (a) checking the sleep effect on motor performance: beneficial, worse or neutral. (b) Dopaminergic adjustment is necessary due to the progression of the nigrostriatal degeneration and the increased sensitivity of the terminals which alter the normal modulator mechanisms of motor centers in LPD patients. Among the many neurotransmitters of the nigro-striatal pathway one can distinguish two with a major influence on REM and non-REM sleep. REM sleep corresponds to an increased cholinergic receptor activity and a decreased dopaminergic activity. This is the reason why REM sleep deprivation by suppressing cholinergic receptor activity ameliorates LPD motor symptoms. L-Dopa and its agonists by suppressing cholinergic receptors suppress REM sleep. L-Dopa has also an arousal effect on Non-REM sleep, repeatedly awakening the patient and enhancing the fragmentation due to the involuntary movements. (c) Socio-physical assistance. (3) The specific therapy consists of: LFS-Sinemet CR, Tolcapone, Intranasal Desmopressin, Domperidon, Cisapride and neurosurgery; SRBD-CPAP, UPPP, nasal interventions, losing weight; RLS-PLM Benzodiazepine (Clonazepam), Opioid, Apomorphine infusion; RBD-Clonazepam and dopaminergic agonists; SRH-Clozapine, Risperidone; SRPD-Nortriptyline, Clozapine, Olanzepine; SA-adjustment; EDS-arousing drugs. Each therapeutic approach must be tailored to the individual LPD patient. PMID- 11489678 TI - The use of estrogen in the treatment of Parkinson's disease. PMID- 11489679 TI - Future of neuroprotection in Parkinson's disease. AB - In Parkinson's disease neuroprotective therapy to rescue dopamine neurons has been proposed. Selegiline is one of neuroprotective drug candidates, as proved by in vivo and in vitro experiments. In this paper, the mechanism underlying neuroprotection by selegiline and related propargylamines was studied against apoptosis induced by an endogenous toxin, N-methyl(R)salsolinol, synthetic 6 hydroxydopamine and peroxynitrite in dopaminergic SH-SY5Y cells. Propargylamines prevented apoptotic DNA damage, through suppression of collapse in mitochondrial membrane potential and following activation of caspase 3 and signal transduction to nuclei. These results suggest that propargylamines may rescue or protect dopamine neurons in Parkinson's disease. PMID- 11489680 TI - The stiff-man syndrome and related disorders. AB - The stiff-man syndrome (SMS) is characterised by rigidity and spasm of predominantly axial and proximal limb muscles. The cause of the condition is unknown but the finding of antibodies to glutamic acid decarboxylase (GAD) in approximately 60% of patients has suggested an autoimmune basis. Pathological findings are limited to a small number of cases which are reviewed in this paper. In some, evidence of an inflammatory aetiology has been found, and there appears to be overlap with progressive encephalomyelitis with rigidity (PER) which may present with a similar clinical picture. The spontaneous muscle activity in SMS and PER is of central origin, related to release of polysynaptic spinal and brainstem reflexes. The SMS is readily distinguished from the continuous muscle activity, spasm and cramps of Isaac's syndrome and neuromyotonia which originate in the peripheral nervous system. Fasciculations, myokymia, myotonia and complex repetitive discharges are characteristic of these peripheral neuromuscular disorders. PMID- 11489681 TI - Perspectives on prostate cancer diagnosis and treatment: a roundtable. AB - This roundtable was held September 30, 2000. It addressed, first of all, the accuracy and proper interpretation of the available prostate-specific antigen assays. Dr. Brawer presented data to demonstrate the specificity of the complexed prostate-specific antigen assay. Dr. Stamey counterpoised evidence that pretreatment prostate-specific antigen levels less than 9 ng/mL are attributable to benign prostatic hyperplasia and therefore are of little value as an indicator of when to initiate treatment for prostate cancer. The other roundtable participants offered reviews and new data regarding hormonal therapy as primary or adjunctive treatment of prostate cancer. Dr. Fowler presented a large retrospective series of men with locally advanced prostate cancer for whom androgen ablation was the primary therapy. Dr. Droller discussed his center's experience in integrating hormonal therapy with brachytherapy. Finally, Dr. Messing reviewed and critiqued the evidence that the combination of hormonal and radiation therapy improves survival. PMID- 11489682 TI - Prognostic assessment of nonmetastatic renal cell carcinoma: a clinically based model. AB - OBJECTIVES: Determining the recurrence risk in patients treated for renal cell carcinoma (RCC) is important for providing prognostic information and planning potential surveillance strategies. The pathologic stage has been the most widely used single prognostic variable. However, with minimally invasive treatment modalities, the pathologic stage may not be readily available. We developed a biostatistical prognostic model for postoperative RCC that is independent of the pathologic stage. METHODS: The records of 296 patients who underwent open nephrectomy for RCC at Johns Hopkins Hospital between 1990 and 1999 were reviewed. Cox proportional hazards regression analysis was used to generate a prognostic model. RESULTS: The recurrence risk (R(rec)) was determined from this model: R(rec)=1.55 x presentation (0-1)+0.19 x clinical size (in centimeters). Using this equation, 79% of patients were identified as low risk compared with 45% of patients considered low risk by pathologic stage (pT1). Moreover, the separation between the high and low-risk survival curves increased. CONCLUSIONS: This model is the first to our knowledge that uses purely clinical variables to assess the postoperative prognosis in patients with RCC. These results, although not validated, provide substantial evidence that preoperative clinical variables may be used instead of the pathologic stage to determine the risk of recurrence. Uncoupling the reliance on pathologic stage for prognostic information removes a potential barrier to novel minimally invasive treatments for renal malignancy and provides a standard to which observation protocols can be compared. In the future, this model may facilitate selection of appropriate patients for less toxic adjuvant or neoadjuvant therapies. PMID- 11489685 TI - Hand-assisted laparoscopic donor nephrectomy minimizes warm ischemia. AB - OBJECTIVES: Traditional open donor nephrectomy is associated with good donor outcomes and excellent allograft function. Laparoscopic donor nephrectomy may accomplish these same goals with less morbidity. We report our initial experience with hand-assisted laparoscopic living donor nephrectomy using a commercially available hand-assist device. METHODS: Donor and allograft outcomes for the first 30 patients undergoing hand-assisted laparoscopic live donor nephrectomy in our institution were prospectively analyzed. RESULTS: Hand-assisted laparoscopic donor nephrectomy was successfully completed in 29 (97%) of 30 donors. Organ dissection was carried out purely laparoscopically. Vessel division and allograft extraction were performed using a hand-assisted technique. The average operative time was 275 minutes (range 193 to 360), with an estimated blood loss of 99 mL (range 50 to 300). Pneumoperitoneum was consistently maintained during the hand assisted portion of the procedure. The mean warm ischemic time was 72.5 seconds (range 30 to 165). On average, the regular diet was resumed after 2.2 days (range 1 to 3), and patients were discharged home 3.4 days (range 2 to 5) after surgery. Eight minor complications occurred in the donor group. Immediate graft function occurred in all 30 cases. No ureteral complications occurred. The recipient creatinine levels ranged from 0.6 to 2.4 mg/dL at an average follow-up of 11.5 months (range 1 to 23). CONCLUSIONS: Laparoscopic donor nephrectomy is technically feasible and can be performed with minimal morbidity. Hand-assisted kidney extraction may help to facilitate immediate allograft function by minimizing the warm ischemic time. PMID- 11489683 TI - Bicalutamide (Casodex) 150 mg as immediate therapy in patients with localized or locally advanced prostate cancer significantly reduces the risk of disease progression. AB - OBJECTIVES: To investigate the efficacy and tolerability of bicalutamide (Casodex) as immediate therapy, either alone or as adjuvant to treatment of curative intent, in patients with localized or locally advanced (T1b-T4, any nodal status, M0) prostate cancer. METHODS: This was a multicenter, prospective, randomized, double-blind, placebo-controlled trial in Europe, South Africa, Australia, and Mexico and is part of the Casodex Early Prostate Cancer program. RESULTS: A total of 3603 men were randomized to receive bicalutamide (n = 1798) or placebo (n = 1805). The patient demographics were well balanced between the two groups. Prior therapy of curative intent had been given to 64% of the patients (prostatectomy [44%], radiotherapy [18%], and prostatectomy and radiotherapy [2%]) and 36% had been monitored with watchful waiting. After a median follow-up of 2.6 years and a median exposure to the study drug of 2.2 years, a significant 43% reduction in the risk of objective progression was observed for the bicalutamide group compared with the placebo group (hazard ratio 0.57, 95% confidence interval 0.48 to 0.69, P << 0.0001). The time to prostate specific antigen doubling was significantly delayed for the bicalutamide group compared with the placebo group (hazard ratio 0.37, 95% confidence interval 0.32 to 0.43, P << 0.001). The survival data were immature, with 7.2% overall mortality. The most frequently reported adverse events with bicalutamide were gynecomastia alone (17.4%), breast pain alone (17.6%), and gynecomastia with breast pain (47.5%). CONCLUSIONS: Bicalutamide 150 mg daily as immediate therapy, alone or as adjuvant to treatment of curative intent, significantly reduced the risk of disease progression in patients with localized or locally advanced prostate cancer. Longer follow-up is underway to assess any benefit in overall survival. PMID- 11489688 TI - Radiographic versus pathologic size of renal tumors: implications for partial nephrectomy. AB - OBJECTIVES: To compare the radiographic size with the pathologic size of renal tumors to determine whether these two measurements vary significantly and to evaluate whether any differences in tumor size could have an impact on the decisions regarding partial nephrectomy. METHODS: In 87 renal tumors excised by partial nephrectomy, the maximum transaxial tumor size on computed tomography (CT) was compared with its corresponding pathologic transverse size. Tumors were locally excised after vascular occlusion and hypothermia. The average size of the tumors selected for partial nephrectomy by preoperative CT scan was 3.4 cm (range 1.9 to 9.3). The difference between the CT size and pathologic size was correlated with the histologic type of the renal tumors. RESULTS: Of the 87 tumors, 52 (60%) were classified as clear cell carcinoma and 35 (40%) as other histologic types (papillary, chromophobe, oncocytoma, and angiomyolipoma). Clear cell carcinomas decreased an average of 0.97 cm versus 0.39 cm for the other tumor types. Of 62 tumors greater than 3 cm on CT, 43 averaged 0.87 cm smaller at pathologic evaluation (36 clear cell and 7 other types) and 19 showed no significant (less than 0.5 cm) decrease in size (2 clear cell and 17 other histologic types). Of 30 tumors greater than 4 cm on CT, 22 clear cell carcinomas shrank more than 1 cm and 8 tumors displaying other histologic features showed no decrease in size. CONCLUSIONS: For renal tumors measuring greater than 4 cm, a decrease in tumor size may help facilitate partial nephrectomy, especially for clear cell carcinomas that do not extensively involve major vascular structures or the collecting system. PMID- 11489689 TI - Predictive values of acute phase reactants, basic fetoprotein, and immunosuppressive acidic protein for staging and survival in renal cell carcinoma. AB - OBJECTIVES: To determine the clinical significance and predictive value of three acute phase reactants (erythrocyte sedimentation rate, C-reactive protein, and ferritin), as well as basic fetoprotein (BFP) and immunosuppressive acidic protein, in patients with renal cell carcinoma. METHODS: Erythrocyte sedimentation rate, C-reactive protein, ferritin, BFP, and immunosuppressive acidic protein levels were measured in 92 patients with renal cell carcinoma diagnosed in 1989 to 1999. The levels were compared with the clinical stage and nuclear grade, and their predictive values of survival were evaluated statistically. RESULTS: All markers, with the exception of BFP, correlated with each other and with the clinical stage and nuclear grade. BFP did not correlate with the acute phase reactants. The log-rank test revealed that the levels of C reactive protein, immunosuppressive acidic protein, and ferritin significantly influenced survival. Multivariate stepwise analysis identified ferritin as the only independent and significant prognostic marker (hazard ratio = 5.624, P = 0.001). However, when age, sex, clinical stage, and nuclear grade were entered into the same analysis, only clinical stage was an independent marker of prognosis. CONCLUSIONS: The results of our study demonstrated that serum ferritin is the most useful marker among five tested factors for staging and predicting survival, although the clinical stage is the best parameter that predicts the prognosis of patients with renal cell carcinoma accurately. PMID- 11489690 TI - Laparoscopic ablation of symptomatic parenchymal and peripelvic renal cysts. AB - OBJECTIVES: To assess the safety and efficacy of laparoscopic ablation of symptomatic renal cysts as minimally invasive therapeutic techniques have largely supplanted open surgical intervention for the treatment of symptomatic renal cysts. METHODS: The records of 32 consecutive adult patients who underwent laparoscopic ablation of renal cysts (11 peripelvic, 21 parenchymal) were retrospectively reviewed. All patients were symptomatic at presentation; 26 had a single cyst, 5 had two cysts, and 1 had four cysts. RESULTS: Twenty patients underwent a transperitoneal laparoscopic approach, and 12 patients underwent a retroperitoneal laparoscopic approach. An average of 3.2 ports were used for each procedure, and no open conversions or transfusions were necessary. When comparing patients with parenchymal and peripelvic cysts, statistically significant differences were noted in the mean operative time (164 versus 233 minutes, respectively; P = 0.003) and mean operative blood loss (98 versus 182 mL, respectively; P = 0.04). Four patients (13%) had complications (one major and three minor), including a persistent ureteral stricture. One patient with negative preoperative aspiration cytology and negative intraoperative frozen section analysis was later found to have malignancy within the cyst wall, necessitating radical nephrectomy and trocar site excision. One patient (3%) developed a recurrence. CONCLUSIONS: Laparoscopic ablation of symptomatic renal cysts is a safe and efficacious procedure. We report an overall complication rate of 13% and a recurrence rate of 3% with a mean follow-up of 18.1 months (median 10.0). PMID- 11489691 TI - Hounsfield unit density in the determination of urinary stone composition. AB - OBJECTIVES: Noncontrast computed tomography (NCCT) has emerged as the diagnostic study of choice in the evaluation of acute flank pain. Recent in vitro studies have suggested that NCCT can be used to predict the composition of urinary stones on the basis of differences in radiodensity (measured in Hounsfield units, HUs). We sought to determine whether the analysis of in vivo urinary stones seen on NCCT could predict their composition. METHODS: Between March 1997 and August 1999, 100 pure stones from patients seen at the Wilford Hall Medical Center in San Antonio, Texas were submitted for analysis. All had been visualized by NCCT before stone passage or retrieval. A General Electric High-Speed Advantage CT scanner evaluated most of these patients by a "flank pain protocol" (ie, helical technique with breath-holding at 120 kV, 200 mA with 5 mm collimation). Each scan was interpreted by one of two staff radiologists who measured the HUs for each stone. A statistical comparison was made between the stone composition and radiodensity. To allow for subsequently observed increases in radiodensity with increasing stone size regardless of composition, the HU value was divided by each stone's largest transverse diameter in millimeters to give the HU density. A statistical comparison was then made between stone composition and HU density. RESULTS: No significant difference was noted between the HU values of calcium oxalate and calcium phosphate stones, and thus they were analyzed collectively as "calcium stones." When the HU values of calcium (n = 87), uric acid (n = 7), struvite (n = 4), and cystine (n = 2) stones were compared, the overlap of ranges precluded accurate identification, and the mean HU values were not significantly different from one another. There was less overlap noted when comparing the HU densities of the stones studied, and no noncalcium stone had an HU density greater than 76 HU/mm. Using one-way analysis of variance, significant differences were noted between the mean HU density of calcium (105 +/- 43) and uric acid (50 +/- 24) stones (P = 0.006). A trend toward significance was found between the mean HU density of the calcium and struvite stones (53 +/- 28, P = 0.073). No significant differences were found among the other stones. CONCLUSIONS: HU density compared with the HU value alone better characterized differences in radiodensities among urinary stones; calcium stones can be distinguished from uric acid stones on the basis of this value. However, neither the HU density nor the mean HU value was able to identify urinary stones in vivo. PMID- 11489692 TI - Is nephroureterectomy necessary in all cases of upper tract transitional cell carcinoma? Long-term results of conservative endourologic management of upper tract transitional cell carcinoma in individuals with a normal contralateral kidney. AB - OBJECTIVES: To evaluate the endoscopic management of upper urinary tract transitional cell carcinoma (TCC) as a first-line treatment in patients with a normal contralateral kidney. METHODS: During an 11-year period, 21 patients diagnosed with upper tract TCC were treated with conservative endourologic techniques using either neodymium:yttrium-aluminum-garnet laser or electrocautery at our institution. The 21 patients were followed up for a mean of 6.1 years (range 1 to 11.6). RESULTS: A total of 8 renal pelvic tumors and 13 ureteral tumors were found. All tumors were Stage T1 or less and grade 3 or less. All tumors were less than 2 cm in the greatest dimension (range 0.4 to 2). Of the 21 patients, 7 (33%) had one local recurrence and 1 (4.7%) developed two local recurrences. Of the 13 ureteral tumors, 6 (46%) recurred; 1 (12%) of the 8 renal pelvic tumors recurred. No recurrent tumor was shown to have an increase in grade. Of the 21 target renal units, 17 (81%) were preserved; 4 (19%) of 21 patients required nephroureterectomy because of tumor recurrence. Overall, 11 patients in the series died, 10 of non-TCC etiology and 1 secondary to invasive bladder TCC that developed after treatment for upper tract TCC. No patients died as a result of conservative management of their upper tract TCC. CONCLUSIONS: Endourologic techniques and conservative treatment of upper tract TCC is an evolving field; however, in properly selected patients, endoscopic treatment can be safely and effectively used as a first-line treatment for upper tract TCC. PMID- 11489693 TI - Use of cadaveric solvent-dehydrated fascia lata for cystocele repair--preliminary results. AB - OBJECTIVES: To present a surgical technique in which cadaveric fascia lata is used for cystocele repair. METHODS: Twenty-one consecutive women (mean age 67 +/- 10 years) with severe cystocele were prospectively enrolled. All patients underwent meticulous clinical and urodynamic preoperative evaluations. Solvent dehydrated, Tutoplast-processed, cadaveric fascia lata was used for cystocele repair. The fascia was anchored transversally between the bilateral arcus tendineus and the cardinal and uterosacral ligaments. Standard endopelvic plication was performed thereafter as a second layer. Patients with overt or occult sphincteric incontinence underwent concomitant pubovaginal sling (PVS) surgery as well, using the same material. The main outcome measures included recurrent urogenital prolapse, persistent or de novo urinary incontinence (stress or urge), and dyspareunia. RESULTS: Of the 21 patients, 19 underwent concomitant PVS, 3 concomitant vaginal hysterectomy, and 8 posterior colporrhaphy in addition to their cystocele repair. The mean follow-up was 20.1 +/- 6.7 months (range 12 to 30). No postoperative complications related to the material or technique occurred. None of the patients developed a recurrent cystocele. Two patients (9%), one of whom underwent concomitant posterior colporrhaphy, developed mild recto-enterocele at 4 to 6 months postoperatively. Six patients underwent concomitant PVS for occult sphincteric incontinence. None developed postoperative stress incontinence. Thirteen other patients underwent concomitant PVS for overt sphincteric incontinence. All but two were stress-continent postoperatively. One half of the patients with preoperative urge or mixed incontinence had persistent urge incontinence postoperatively. None of the patients developed postoperative de novo urge incontinence or dyspareunia. CONCLUSIONS: The use of solvent dehydrated cadaveric fascia lata for cystocele repair, as well as PVS, is associated with encouraging short and medium-term results. Long-term follow-up is needed to evaluate whether these results are durable. PMID- 11489694 TI - Psoas hitch ureteral reimplantation in adults--analysis of a modified technique and timing of repair. AB - OBJECTIVES: The psoas hitch ureteral reimplantation technique has been used with great success to bridge defects in ureteral length due to injury or planned resection. Several surgical principles have been historically stressed when performing this procedure, including adequate mobilization of the bladder, fixation of the bladder to the psoas tendon before reimplantation, the use of a submucosal nonrefluxing-type ureteral anastomosis, and a 6-week delay before attempting repair after a surgical injury. We retrospectively reviewed patients who underwent ureteroneocystostomy with a psoas hitch, evaluated the relevance of these principles, and describe a modification of the technique. METHODS: All patients undergoing psoas hitch ureteral reimplantation were reviewed. The indications, complications, and long-term outcomes were assessed. RESULTS: Between 1989 and 1999, 24 patients underwent psoas hitch reimplantation at our institution. The indications were operative injury in 11, planned surgical resection during nonurologic pelvic surgery in 4, cancer in 4, stricture in 4, and trauma in 1. Refluxing-type ureteral anastomoses were performed in 17 cases. One case of postoperative urosepsis occurred. A delayed repair after operative injury did not improve the operative time or overall morbidity. No cases of chronic flank pain, recurrent pyelonephritis, persistent severe hydronephrosis, or compromised renal function, as measured by a change in baseline serum creatinine level, occurred. No patient required reoperation for either early or delayed complications or failure of the repair at a follow-up of 1 to 122 months (mean 32.75). CONCLUSIONS: Psoas hitch ureteral reimplantation is an effective means of treating defects in ureteral length. Immediate repair may be safely undertaken as soon as the ureteral injury is recognized. Long-term sequelae are unusual in adults, even when using refluxing-type ureteral anastomoses. PMID- 11489695 TI - One 24-hour frequency-volume chart in a woman with objective urinary motor urge incontinence is sufficient. AB - OBJECTIVES: To determine whether one 24-hour frequency-volume (FV) chart in a woman with objective urinary motor urge incontinence is sufficient to gain insight into her voiding habits. METHODS: Retrospectively, women 18 years old and older with urodynamically confirmed urinary motor urge incontinence without neurologic disease or other urologic pathologic findings, who correctly completed two or three 24-hour FV charts during normal daily life, were studied. For each patient, we compared the voiding parameters (mean voided volume, diuria, nocturia, minimum and maximum voided volumes, and total voided volume) and fluid intake calculated from the first FV chart with the same data calculated from all the FV charts. RESULTS: Ninety-eight adult women were recruited. A marked dropout occurred, and 60 patients remained to be evaluated (mean age 54.5 +/- 15.5 years). The calculated voiding parameters agreed with those in published reports. Strong associations existed between all voiding parameters and fluid intake calculated from the first FV chart and those calculated from all FV charts (P <0.001 for all parameters). Compared with the well-known variability, the differences between the voiding parameters calculated from the first FV chart and those from all charts were small. These results agreed with studies in other patient groups (benign prostatic hyperplasia and interstitial cystitis) that analyzed the use of only one 24-hour FV chart. CONCLUSIONS: The use of only one 24-hour FV chart completed during normal daily life by a woman with objective urinary motor urge incontinence is sufficient to gain insight into her voiding habits. PMID- 11489696 TI - Effect of levator ani muscle contraction on urethrovesical and anorectal pressures and role of the muscle in urination and defecation. AB - OBJECTIVES: To investigate the effect of levator ani muscle (LAM) (the main muscle in the pelvic floor) contraction on the anorectal and urethrovesical pressures and elucidate its role in the mechanisms of defecation and urination. METHODS: In 18 healthy volunteers (10 men and 8 women; mean age 38.6 +/- 10.2 years), the LAM was stimulated to contract by a concentric needle electrode, and the anal, rectal, urethral, and vesical pressure responses were recorded. The test was repeated once after LAM anesthetization with Xylocaine injection and again after saline instead of Xylocaine injection into the LAM. RESULTS: On LAM stimulation, the anal and urethral pressures had a significant decline (both P <0.05), and the rectal and vesical pressures exhibited no significant changes (both P >0.05). The anal, rectal, urethral, and vesical pressures did not show significant changes on stimulation of the anesthetized LAM; after saline injection, the pressure response was similar to that before injection (P >0.05). These results were reproducible. CONCLUSIONS: LAM contraction lowers the anal and urethral pressures and appears to assist in the evacuation process. We therefore propose that the LAM is a muscle of evacuation and that LAM dysfunction could lead to defecation and urination disorders. PMID- 11489699 TI - Prevalence of chronic prostatitis in men with premature ejaculation. AB - OBJECTIVES: To investigate the prevalence of chronic prostatitis in men with premature ejaculation. The etiology of premature ejaculation is currently considered psychological in nature. However, the possibility that urologic, hormonal, or neurologic factors may contribute to this condition should be considered in its management. METHODS: We evaluated segmented urine specimens before and after prostatic massage and expressed prostatic secretion specimens from 46 patients with premature ejaculation and 30 controls by bacteriologic localization studies. The incidence of premature ejaculation in the subjects with chronic prostatitis was also evaluated. RESULTS: Prostatic inflammation was found in 56.5% and chronic bacterial prostatitis in 47.8% of the subjects with premature ejaculation, respectively. When compared with the controls, these novel findings were statistically significant (P <0.05). CONCLUSIONS: Considering the role of the prostate gland in the mechanism of ejaculation, we suggest a role for chronic prostate inflammation in the pathogenesis of some cases of premature ejaculation. Since chronic prostatitis has been found with a high frequency in men with premature ejaculation, we stress the importance of a careful examination of the prostate before any pharmacologic or psychosexual therapy for premature ejaculation. PMID- 11489700 TI - Discontinuation of alpha-blockade after initial treatment with finasteride and doxazosin in men with lower urinary tract symptoms and clinical evidence of benign prostatic hyperplasia. AB - OBJECTIVES: To determine the effect of discontinuation of alpha-blockade and continuation of finasteride in men with lower urinary tract symptoms and enlarged prostates receiving combination treatment and to determine whether the alpha blocker dose influences the ability to discontinue it. METHODS: We treated 272 consecutive men with a prostate size greater than 40 g and American Urological Association symptom score greater than 20 with 5 mg finasteride and 2 mg doxazosin daily. Two hundred forty men reported a favorable response to therapy, defined as any reduction in symptom score and toleration of the medications. The dose was maintained at 2 mg doxazosin in 100 men and was titrated to 4 mg doxazosin in 80 patients and to 8 mg doxazosin in an additional 60 patients. We discontinued doxazosin at 3, 6, 9, or 12 months, while continuing finasteride, and then re-evaluated the patients 1 month later to determine whether any worsening of symptoms had resulted. RESULTS: In patients discontinuing doxazosin at 3 months, success (defined as no increase in symptom score and no desire to resume doxazosin) was reported by 20%, 15%, and 13% of those taking 2, 4, and 8 mg, respectively. In patients discontinuing doxazosin at 6 months, success was reported by 48%, 45%, and 40% of those taking 2, 4, and 8 mg, respectively. In patients discontinuing doxazosin at 9 months, success was reported by 84%, 80%, and 73% of those taking 2, 4, and 8 mg, respectively. In patients discontinuing doxazosin at 12 months, success was reported by 84%, 85%, and 87% of those taking 2, 4, and 8 mg, respectively. CONCLUSIONS: Patients with lower urinary tract symptoms and moderately enlarged prostates initially receiving combination therapy using finasteride and an alpha-blocker are likely to experience no significant symptom deterioration after discontinuing the alpha-blocker after 9 to 12 months of combination therapy regardless of the dose of alpha-blocker chosen. PMID- 11489704 TI - Impact of a modified apical dissection during radical retropubic prostatectomy on the occurrence of positive surgical margins: a comparative study in 212 patients. AB - OBJECTIVES: To assess the impact of a modified technique of apical dissection during radical retropubic prostatectomy on the occurrence of positive surgical margins (PSMs). METHODS: Between 1992 and 1998, 212 nonconsecutive patients with localized prostate cancer (57 T1, 155 T2) underwent radical retropubic prostatectomy and were divided into two groups: group 1, 85 patients who underwent surgery before January 1994; and group 2, 127 patients who underwent surgery with the modification of the apical dissection after January 1994. The modified technique consisted of a wide excision of periprostatic soft tissue at the apex, including the bilateral neurovascular bundles. The clinical data (age, prostate-specific antigen, clinical staging) and pathologic findings (pathologic staging, Gleason score, PSM rate) of the two groups were compared. RESULTS: No significant difference was found between the two groups regarding the median prostate-specific antigen level (10.8 ng/mL and 9.5 ng/mL), Gleason score, and pathologic staging. Overall, the PSM rate was 53% in group 1 and 20.5% in group 2 (P <0.001). The number of PSMs decreased 2.6-fold in group 2. The PSM rate was significantly reduced at the apex with the new technique (group 1, 33.3%; group 2, 7.7%; P = 0.008). The PSM rate expressed with the odds ratio was 4.4-fold lower for patients in group 2 than for those in group 1. CONCLUSIONS: The modified apical dissection in radical retropubic prostatectomy significantly improves the PSM rate in patients with localized T1-T2 prostate cancer. PMID- 11489703 TI - Clinical predictors of spontaneous acute urinary retention in men with LUTS and clinical BPH: a comprehensive analysis of the pooled placebo groups of several large clinical trials. AB - OBJECTIVES: To comprehensively evaluate clinical predictors of spontaneous acute urinary retention (AUR) across pooled data of placebo-treated patients from clinical trials conducted in men with lower urinary tract symptoms and clinically diagnosed benign prostatic hyperplasia. METHODS: Data from the placebo-treatment groups of several prospective, randomized clinical trials conducted in the United States (n = 3040), Scandinavia, Canada, and worldwide (n = 2295) were combined in the analyses. More than 110 variables were considered individually and in combination as predictors of AUR using logistic regression analysis and classification and regression tree methods with a split-sample approach to cross validation. RESULTS: The different methods of analysis identified consistent potential predictors of episodes of AUR. When prostate volume was included in the analyses, it was selected as the initial variable discriminating men with and without subsequent AUR. Omitting prostate volume because of its availability in only a subset of men, a logistic model including serum prostate-specific antigen (PSA), urinating more than every 2 hours, symptom problem index, maximum urinary flow rate, and hesitancy of urination had good predictive properties (area under the receiver-operating characteristic curve [AUC] = 0.742 +/- 0.047), as did a model with PSA (AUC = 0.716 +/- 0.045). A classification and regression decision tree with the same variables predicted AUR (AUC = 0.74, sensitivity = 72%, specificity = 67%) as well as did a tree with PSA alone (AUC = 0.70, sensitivity = 75%, specificity = 64%). CONCLUSIONS: Prostate volume and serum PSA are strong predictors of AUR in placebo-treated men with lower urinary tract symptoms and clinically diagnosed benign prostatic hyperplasia who were screened for prostate cancer. From more than 110 variables, logistic models and decision trees with PSA alone were comparable to expanded models that included PSA, urinary frequency and hesitancy, flow rate parameters, and symptom problem index, and to a scoring algorithm. PMID- 11489705 TI - Volume-specific cutoffs are necessary for reproducible application of prostate specific antigen density of the transition zone in prostate cancer detection. AB - OBJECTIVES: To determine the effect of prostate volume on the specificity of prostate-specific antigen density (PSAD) and PSAD of the transition zone (PSA-TZ) in the detection of prostate cancer. METHODS: Between February 1994 and April 1998, transrectal ultrasound-guided prostate needle biopsies were performed in 235 men with serum prostate-specific antigen (PSA) levels between 4.0 and 10.0 ng/mL. The PSAD and PSA-TZ specificities were calculated at 95% sensitivity cutoff levels generated from the whole group, as well as from cohorts stratified by transition zone index or prostate volume. RESULTS: Statistical significance was noted between the benign (n = 176) and prostate cancer (n = 59) groups for all tested PSA parameters. At 95% sensitivity, PSA-TZ carried a specificity of 37.5% compared with 29.6% for PSAD. When applying a single 95% sensitivity cutoff derived from the entire group to individual volume-stratified cohorts, the specificity decreased to 0% in glands less than 30 g in size. A 95% sensitivity PSA-TZ cutoff generated individually for volume-stratified cohorts of glands less than 30, 30 to 40, and 40 to 60 g resulted in more consistent specificity of 28.2%, 35.2%, and 45.7% for each cohort, respectively. CONCLUSIONS: Unlike whole group-derived cutoffs, the use of volume-specific PSA-TZ cutoffs allows consistently high specificity in all volume-stratified cohorts. The discrepancies in the PSA-TZ and PSAD specificities in published reports are likely due to the application of published cutoffs to populations of differing prostate volumes. The use of volume-specific cutoffs results in reproducible specificity in populations with differing prostate volume distribution, and thereby definitively resolves the differences in PSA-TZ specificity reported in published reports. PMID- 11489708 TI - Assessment of the risk of positive surgical margins with pelvic phased-array magnetic resonance imaging in patients with clinically localized prostate cancer: a prospective study. AB - OBJECTIVES: We assessed magnetic resonance imaging (MRI) performance in the prediction of positive surgical margins (PSMs) before radical prostatectomy in a prospective study correlating the MRI results and pathologic findings. METHODS: Between January 1995 and December 1999, 176 patients (mean age 64.2 years, range 49 to 75), with localized prostate cancer (49 with Stage T1 and 127 with Stage T2) underwent preoperative MRI with a pelvic phased-array coil (Tesla-1, Siemens) at a mean interval of 35 days after randomized transrectal biopsies. The mean preoperative prostate-specific antigen level was 10.9 ng/mL (range 1.2 to 39). The MRI studies and specimen analysis were performed by one radiologist unaware of the clinical and biopsy findings and by one pathologist, respectively. Multivariate analysis was performed to compare the predictive value of MRI staging, prostate-specific antigen value, and preoperative Gleason score to identify the PSM rate. RESULTS: Of the 176 patients, 131 (74%) had Stage T2 disease by MRI and 45 (26%) Stage T3 disease by MRI. Pathologic staging showed 103 with pT2 and 73 with pT3. Overall, the PSM rate of the series was 18%. The PSM rate was 13.7% and 31% for patients with T2 and T3 disease by MRI, respectively. For the T3 MRI cases, the PSM rate was 2.32-fold higher. MRI staging, like the prostate-specific antigen value, was a predictive factor of PSMs (P = 0.05). CONCLUSIONS: The results of this study show that preoperative MRI staging with the phased-array coil may be helpful in predicting the PSM risk in radical prostatectomy candidates with clinically localized prostate cancer. PMID- 11489709 TI - Addition of radiation therapy to androgen ablation improves outcome for subclinically node-positive prostate cancer. AB - OBJECTIVES: To determine the outcome for node-positive prostate cancer treated by early androgen ablation with or without prostatic radiation. METHODS: Two hundred fifty-five men with lymphadenectomy-proven pelvic nodal metastases treated with early androgen ablation alone (n = 183) or with combined ablation and radiation (n = 72) between 1984 and 1998 were retrospectively reviewed for disease outcome and survival. Post-treatment disease status was based on the prostate-specific antigen levels or on the clinical and radiographic status for patients treated before 1987. Univariate and multivariate statistics were used to determine the prognostic factors and assess the influence of radiation treatment. RESULTS: With a median follow-up of 9.4 years, the 5, 10, and 13-year overall survival rate for those treated with early ablation alone was 83%, 46%, and 21%, respectively. The freedom from relapse or rising prostate-specific antigen rate for these patients was 41%, 25%, and 19% at 5, 10, and 13 years, respectively. Distant metastasis and local recurrence occurred with a 10-year actuarial incidence of 44% and 51%, respectively. With a median follow-up of 6.2 years, the 5 and 10-year overall survival rate for those treated with radiation and ablation was 92% and 67%, respectively. The freedom from relapse or rising prostate-specific antigen rate in these men was 91% and 80% at 5 and 10 years, respectively. The superior outcome for combined ablation and radiation was substantial and statistically significant in the univariate and multivariate analyses. CONCLUSIONS: Early androgen ablation alone has little curative potential for node-positive prostate cancer. The addition of prostatic radiation to ablation resulted in substantial and significant improvement in disease control and patient survival. PMID- 11489710 TI - Intermittent androgen deprivation: update of cycling characteristics in patients without clinically apparent metastatic prostate cancer. AB - OBJECTIVES: To update the cycling characteristics and patterns of treatment in patients receiving intermittent androgen deprivation (IAD) for clinically localized and recurrent prostate cancer. METHODS: We report our experience with 61 patients treated with IAD. Thirty-four patients had received no prior treatment, and 27 had developed recurrent disease after previous local therapy. No patient had clinically apparent metastatic disease before the initiation of therapy. The mean and median serum prostate-specific antigen (PSA) level before treatment was 25.3 ng/mL and 16.0 ng/mL, respectively (range 0.5 to 190 ng/mL). For each cycle, androgen deprivation was continued until PSA became undetectable or a nadir level was reached. Patients were then observed without treatment, and therapy was reinstituted after the serum PSA value reached a predetermined level. Patients were no longer eligible to cycle off treatment when their serum PSA increased despite ongoing androgen deprivation or if any objective evidence of disease progression was present on imaging studies. RESULTS: Follow-up ranged from 7 to 60 months (mean 30) from the start of treatment. Patients received from one to five treatment cycles (median two), with a median cycle length of 14 months. The median nadir serum PSA level during androgen deprivation was 0.01 ng/mL and was reached within an average of 6 months (range 4 to 9) after beginning treatment. Patients spent an average of 45% of the time not receiving therapy, but the time off therapy decreased as the number of treatment cycles increased. Five patients (8.1%) demonstrated progressive disease, with a median time to progression of 48 months. When examining the cycling characteristics of these patients, no consistent pattern of failure emerged. CONCLUSIONS: IAD appears to be a viable treatment option in select patients with localized prostate cancer. With each consecutive cycle, the amount of time the patient was not receiving therapy decreased, despite achieving a low nadir PSA. Longer follow up with more patients failing IAD will be required before clear patterns of failure emerge in these patients. PMID- 11489711 TI - Neurophysiologic testing in chronic pelvic pain syndrome: a pilot study. AB - OBJECTIVES: Studies of male chronic pelvic pain syndrome (CPPS) have generally centered on the pathologic features of the prostate rather than on the neurology of pain. Electrodiagnostic studies examine the integrity of somatosensory nerve pathways consisting of large, group A fibers. Heat sensation and visceral (autonomic) pain is mediated through small, unmyelinated C fibers, which can be tested cutaneously by thermal sensory analysis. We hypothesized that CPPS pain is mediated by these small C fibers. METHODS: All subjects and controls had no history of neurologic disease and had normal neurologic examinations. Phase I: 14 patients with CPPS underwent electrodiagnostic testing using pudendal somatosensory evoked potentials and bulbocavernosus reflex latency measurements. Phase II: 31 patients with CPPS and 14 controls underwent thermal sensory analysis testing on the perineum and anterior thigh using noxious heat stimuli. Subjects used a computer-generated visual analog scale to dynamically report their discomfort. The peak and slope of the computer-generated visual analog scale were analyzed. RESULTS: Phase I: two patients had delayed latency of the somatosensory evoked potentials, but additional evaluation with magnetic resonance imaging revealed no definable lesion. Phase II: with thermal sensory analysis, men with CPPS reported higher intensity pain at lower temperatures (P = 0.03). Men with CPPS also had higher peak computer-generated visual analog scale scores on perineal testing. No difference in thermal testing on the anterior thigh was found between the two groups. CONCLUSIONS: Large, myelinated somatic fibers do not play a significant role in the pathophysiology of CPPS. Patients with CPPS have an altered sensation of perineal pain elicited by heat, which may represent a C-fiber-mediated effect. PMID- 11489712 TI - Suppression of spermatogenesis in ipsilateral and contralateral testicular tissues in patients with seminoma by human chorionic gonadotropin beta subunit. AB - OBJECTIVES: The pathologic complexity of the testicular tumor makes it difficult to demonstrate exactly the relationship between the impaired spermatogenesis in patients with a testicular tumor and the serum level of the human chorionic gonadotropin beta subunit (beta-hCG). Therefore, we performed quantitative evaluation of spermatogenesis in ipsilateral and contralateral testicular tissues of seminoma to simplify the relation pathologically and endocrinologically and to demonstrate the exact correlation between spermatogenesis and serum beta-hCG levels. METHODS: Fifty-three biopsy specimens from ipsilateral and contralateral testicular tissues of seminoma were analyzed histologically. The quantitative evaluation of spermatogenesis was performed by the mean Johnsen's score count (MJSC). Beta-hCG expression in seminoma was examined immunohistochemically. Serum beta-hCG, testosterone, estradiol, luteinizing hormone, and follicle-stimulating hormone levels were analyzed before orchiectomy. RESULTS: A significant linear relationship (r = -0.82; P <0.005) was found between the serum level of beta-hCG and the MJSC in contralateral testicular tissues but not in ipsilateral ones, although the suppression of spermatogenesis was observed in both sides without suppression of luteinizing hormone and/or follicle-stimulating hormone production. CONCLUSIONS: A clearcut fall in the MJSC with an associated rise in the serum level of beta-hCG was demonstrated in the contralateral testicular tissues but not in the ipsilateral ones of seminoma. It seems most likely that serum beta-hCG suppresses spermatogenesis in both ipsilateral and contralateral testicular tissues without the suppression occurring through the hypothalamus pituitary-gonadal system, and also that some less well recognized factors affect spermatogenesis, making the relation between serum beta-hCG and MJSC obscure in ipsilateral testicular tissues. PMID- 11489713 TI - Biologic variability of sperm DNA denaturation in infertile men. AB - OBJECTIVES: To examine sperm DNA denaturation (DD) in fertile and infertile men and assess the variability of conventional semen parameters and sperm DD in repeated semen samples from infertile men. METHODS: Twenty-one consecutive nonazoospermic, infertile men each submitted two semen samples, 2 to 6 weeks apart. We examined semen samples from consecutive fertile men (n = 10) presenting for vasectomy as controls. Standard semen parameters (World Health Organization criteria) and sperm chromatin structure (evaluated by flow cytometry analysis of acridine orange-treated spermatozoa and expressed as the percentage of spermatozoa with denatured DNA) were monitored. RESULTS: Fertile men had a significantly higher sperm concentration and percentage of sperm motility and a significantly lower percentage of sperm with DD than did infertile men (36 +/- 5.2 x 10(6)/mL versus 12.5 +/- 2.2 x 10(6)/mL, 60.0% +/- 5.2% versus 30.1% +/- 4.1%, and 8.9% +/- 1.9% versus 20.3% +/- 2.5%, respectively, P <0.05). The sperm concentration, sperm motility, and percentage of spermatozoa with DD were not significantly different between the first and second semen samples from the infertile men. Sperm DD showed the lowest average within-subject coefficient of variation (SD/mean), followed by motility and concentration (coefficient of variation 21%, 24%, and 35%, respectively). CONCLUSIONS: Our data demonstrate that infertile men have significantly higher sperm DD compared with fertile men and that sperm DD exhibits a low coefficient of variation ( approximately 20%) on repeated assessment. These data suggest that sperm DD has a relatively low degree of biologic variability. PMID- 11489714 TI - Analysis of the consistency of intraurethral prostaglandin E(1) (MUSE) during at home use. AB - OBJECTIVES: To determine the consistency of a successful response to intraurethral prostaglandin E(1) (MUSE), an effective treatment for a proportion of patients with erectile dysfunction, during at-home use in men who had a successful response in the office, to define the factors that correlated with the consistency of the response, and to determine whether patients continued long term use. METHODS: The study group consisted of men with documented erectile dysfunction who had a successful response (grade 3 or 4 erection) to MUSE during an initial in-office dose. All enrollees completed a home diary assessing the treatment success during at least five medication administrations. Factors such as patient age, vascular risk factor status, and degree of in-office response were prospectively evaluated as correlates of response consistency. Finally, patient follow-up was conducted in an attempt to define how many patients were continuing to use this medication as a long-term treatment modality. RESULTS: Two hundred twelve patients underwent screening and 72 (34%) patients had in-office success. The overall per patient at-home consistency rate was 51%. None of the aforementioned factors correlated with the consistency of the response. At a mean of 9 months after beginning treatment, only 31% of the in-office responders were continuing to use MUSE. CONCLUSIONS: Although MUSE is an appropriate and safe treatment for many patients with erectile dysfunction, the lack of consistency is a significant consideration. Clinicians should pay close attention to patient education in an effort to promote realistic expectations for MUSE therapy. Furthermore, the lack of consistency may significantly lower the patient's interest in continuing this treatment long term. PMID- 11489715 TI - Outcomes of one-stage techniques for proximal hypospadias repair. AB - OBJECTIVES: To evaluate the success of three preputial flap techniques in the one stage correction of proximal hypospadias. METHODS: From 1993 to 1999, 43 children underwent one-stage proximal hypospadias repair using preputial flaps, which were performed by a single surgeon. Of the 43 patients, 21 (48.8%), 10 (23.2%), and 12 (27.9%) underwent tubularized island flap urethroplasty, double-faced tubularized island flap urethroplasty, and onlay island flap urethroplasty, respectively. The age range of the patients at surgery was 18 months to 14 years (mean 3.4). Dorsal plication was required for chordee repair in 12 patients (3 in tubularized island flaps, 3 in double-faced tubularized island flaps, and 6 in onlay island flaps). In addition to the dorsal plication, posterior dissection of the urethral plate without division was performed on 3 of the 6 patients with mild to severe chordee in the onlay island flap group. The incidence of complications requiring repeated surgery was evaluated for each group. RESULTS: The follow-up was 8 months to 7 years (mean 4.1 years). The overall complication rate was 90% for the double faced tubularized island flap repair, 38% for the tubularized island flap repair, and 33% for the onlay island flap repair. Recurrent chordee was observed in 2 (66.6%) of the 3 patients who underwent onlay island flap repair with urethral plate dissection. CONCLUSIONS: The use of a tubularized island flap is suggested for one-stage repair of proximal hypospadias, especially in the patients with severe chordee. Because of the high complication rates, the double-faced tubularized island flap technique is not advocated. The onlay island flap may also be used for proximal hypospadias repair if mild chordee is present. Because of the high recurrent chordee rate, dissection of the urethral plate without division is not suggested in the onlay island flap technique. PMID- 11489717 TI - Subcutaneous frenulum flap (SCUFF) for iatrogenic or primary megameatus and reoperative hypospadias repair. AB - OBJECTIVES: Megameatus, whether primary or iatrogenic, can be managed by one of several tubularization techniques. Repair after prior circumcision or after failed hypospadias repair can occasionally be challenging because of the lack of local vascularized tissues. The use of an inferiorly based frenulum flap as a buttressing layer for these unusual repairs is described. METHODS: An inferiorly based island flap of frenulum skin is de-epithelialized and advanced over the urethral repair. RESULTS: The procedure has been used in 5 cases: two reoperative hypospadias repairs, one primary megameatus repair, and two repairs of traumatic fistula/megameatus after circumcision. Overall, the cosmetic results were excellent, with one postoperative fistula. CONCLUSIONS: This is a straightforward technique providing added vascularity in these relatively unusual cases in which a dorsal or meatal-based flap is not convenient or feasible. PMID- 11489718 TI - Preputial flap for primary closure after excision of tumors on the glans penis. AB - In patients with large defects of the glans penis consequent to organ-preserving tumor excision, we describe a vascularized flap formed from the outer preputial leaf for primary defect coverage. We have used this successfully in 3 men in whom both erections and penile length have been preserved. PMID- 11489719 TI - Retained laser fiber ureteral calculus. PMID- 11489720 TI - Ureteral triplication with ectopic upper pole moiety. PMID- 11489721 TI - Laparoscopic ureteral reimplantation for ureteral lesion secondary to transvaginal ultrasonography for oocyte retrieval. AB - Transvaginal ultrasound-guided follicular puncture for oocyte retrieval is a highly efficient and minimally invasive method for assisted reproductive techniques. Complications related to this procedure are rare. We report the case of a ureteral stricture secondary to ultrasound-guided follicular puncture for oocyte retrieval that was corrected by a laparoscopic approach. This approach can minimize postoperative pain, the length of hospitalization, and the period of convalescence and should be considered a minimally invasive option in the management of this rare complication of oocyte retrieval. PMID- 11489722 TI - Primary localized amyloidosis of the ureter. AB - Primary amyloidosis is a rare condition that can involve the urinary tract. These lesions can occur anywhere in the collecting system and are often mistaken clinically for malignancies. We report a case of localized ureteral amyloidosis. Our treatment consisted of a conservative approach with local resection. PMID- 11489723 TI - Acute bilateral renal cortical necrosis as a cause of postoperative renal failure. AB - Acute renal failure after a major intra-abdominal operation is, unfortunately, not an infrequent occurrence. Acute tubular necrosis, the most common cause of postoperative renal failure, usually follows a predictable clinical course, with most patients recovering full renal function. We describe a patient who developed acute renal failure after orthotopic liver transplantation. Subsequent workup revealed the patient to have acute bilateral renal cortical necrosis. Bilateral renal cortical necrosis is an extremely rare cause of renal failure and an even rarer cause of postoperative renal failure. We discuss the diagnosis and management of this uncommon disorder and review the salient literature. Of the approximately 15 known reported cases involving native kidneys after a major nonobstetric abdominal operation in the world literature, we believe this is the first to be related to an orthotopic organ transplant. PMID- 11489724 TI - Expanding the differential diagnosis of the acute scrotum: ventriculoperitoneal shunt herniation. AB - An 18-month-old boy presented to the emergency department after 4 hours of inconsolability and acute scrotal swelling. The physical examination revealed a new scrotal hydrocele with migration of a ventriculoperitoneal shunt into the right hemiscrotum. The presence of a ventriculoperitoneal shunt has been associated with increased patency of the processus vaginalis and scrotal hydroceles. The presentation of an acute scrotum in a child with a ventriculoperitoneal shunt should be recognized as a possible shunt complication. Migration of the shunt through the processus vaginalis is an extremely uncommon event. PMID- 11489725 TI - Diagnostic difficulties before definitive treatment of an extragonadal retroperitoneal germ cell tumor. AB - A primary extragonadal germ cell tumor of the retroperitoneum was diagnosed in a 47-year-old man without elevated serum alpha-fetoprotein, human chorionic gonadotropin, or lactate dehydrogenase levels. The diagnosis was made by histologic analysis after resection. The patient responded well to a combination of cisplatin, etoposide, and ifosfamide, achieving a partial response with four cycles. Residual tumor resection revealed necrotic tissue only. The patient was alive and disease free 24 months after diagnosis. The diagnostic difficulties of this particular situation are discussed. PMID- 11489726 TI - A case of familial clustering of interstitial cystitis and chronic pelvic pain syndrome. AB - A case of familial clustering of interstitial cystitis (IC) and chronic pelvic pain syndrome (CPPS), a symptom complex similar to IC that occurs in men, is reported. The proband was a 28-year-old woman with a 6-month history of severe frequency, urgency, and genital pain. After cystoscopy with hydrodistention and biopsy, a diagnosis of IC was made. IC was also diagnosed in the patient's mother and in two of her brothers, previously considered to have CPPS (category IIIB CPPS). A third brother was asymptomatic. This case highlights the importance of genetic factors in the onset of symptoms and natural history of IC and CPPS. PMID- 11489727 TI - Penile curvature associated with scleroderma. AB - We present 2 cases of Peyronie's disease in patients with scleroderma. This association is uncommon, and our patients' symptoms and disease course differed from that typical for Peyronie's disease. We suggest a more conservative approach when treating patients whose penile curvature is associated with a systemic disease such as scleroderma. PMID- 11489728 TI - Modulation of omega-3/omega-6 polyunsaturated ratios with dietary fish oils in men with prostate cancer. AB - OBJECTIVES: The results of epidemiologic and animal studies support the role of a low-fat diet supplemented with omega-3 fatty acids contained in fish oil in preventing the development and progression of prostate cancer. As a first step in studying the role of a low-fat, fish oil-supplemented (LF/FOS) diet in a clinical setting, we conducted a prospective study in men with untreated prostate cancer to evaluate whether a 3-month dietary intervention affects the ratio of omega-3 to omega-6 fatty acids in plasma and gluteal fat. In addition, we evaluated the feasibility of studying cyclooxygenase-2 (COX-2) expression in serial prostate needle biopsy specimens before and after the diet. METHODS: Nine men with untreated prostate cancer consumed an LF/FOS diet for 3 months. Plasma, gluteal adipose tissue, and prostate needle biopsy specimens were obtained from each patient before and after the intervention. The fatty acid compositions of the plasma and gluteal adipose tissue were determined by gas-liquid chromatography, and the COX-2 expression in the prostatic tissue specimens was determined by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Short-term intervention with an LF/FOS diet caused a significant increase in the omega-3/omega-6 fatty acid ratio in plasma (P = 0.002) and gluteal adipose tissue (P = 0.002) in men with prostate cancer. The COX-2 expression in prostatic tissue was quantitated by RT-PCR in 7 of 9 patients, and COX-2 expression decreased in 4 of these 7 patients. CONCLUSIONS: A short-term dietary intervention in men with prostate cancer leads to a significant increase in the omega-3/omega-6 fatty acid ratios in plasma and adipose tissue. The potential for this diet to prevent the development and progression of prostate cancer by way of altered COX-2 expression and prostaglandin production in prostatic tissue requires further study. PMID- 11489729 TI - Different expression of androgen receptor coactivators in human prostate. AB - OBJECTIVES: To investigate the expression of androgen receptor (AR) coactivators in the human prostate for a better understanding of androgen action in prostate cancer. METHODS: Using reverse transcriptase-polymerase chain reaction, we examined the expression levels of AR coactivators (ARA55, SRC1, ARA54, TIF2, RAC3) in four prostate cancer cell lines (DU145, PC3, LNCaP, and LN-TR2), nine benign prostatic tissue samples, and 21 prostate cancer tissue specimens. RESULTS: In the cell lines, SRC1 was expressed ubiquitously at almost equal amounts. Contrary to this, ARA55, ARA54, TIF2, and RAC3 displayed cell line specific expression. In the LN-TR2 cells, established from LNCaP cells by long term treatment with tumor necrosis factor-alpha, the expression levels of ARA55 and TIF2 were much higher than those in the LNCaP cells. In every prostatic tissue specimen, the expression levels of TIF2 and RAC3 were very low. The expression levels of ARA55 and SRC1 were higher in the cancer specimens with a higher grade or poor response to endocrine therapy than in those with a lower grade or good response to endocrine therapy. CONCLUSIONS: Prostate cancer cells express AR coactivators. Long-term stimulation by tumor necrosis factor-alpha could increase ARA55 and TIF2 expression in LNCaP cells. The different expression of coactivators may contribute to the different response of prostate cancer to androgenic stimulation or endocrine therapy. PMID- 11489730 TI - Increased blood flow after catheterization and drainage in the chronically obstructed rabbit urinary bladder. AB - OBJECTIVES: To determine the effect of drainage on rabbit bladder blood flow after 4 weeks of partial outlet obstruction. Previous studies have shown that catheterization and drainage of the urinary bladder in control rabbits resulted in a significant nitric oxide-induced increase of blood flow to the bladder. It was also shown that 4 weeks' partial outlet obstruction caused a significant decrease in blood flow to the bladder. METHODS: Male New Zealand White rabbits underwent partial outlet obstruction by standard methods. After 4 weeks, the blood flow to the bladder muscle and mucosa was determined by a microsphere technique. Within 1 to 2 minutes after transurethral catheterization and complete drainage of the bladder, the blood flow was again determined. Unobstructed animals served as controls. Four other control animals underwent a repetitive blood flow study during 10 minutes to determine the time frame of blood flow changes after drainage. Blood flow was also measured in 2 control rabbits after transurethral catheterization without drainage and in 2 control rabbits after drainage by suprapubic puncture. To exclude the possibility that increased intravesical pressure alters the blood flow measurements, the relationship between the intravesical volume and the bladder pressure was examined in the obstructed rabbits. RESULTS: After drainage of the bladder, the blood flow to the bladder muscle increased 4.5-fold in the decompensated obstructed group (bladder weights greater than 15 g) and 2.5-fold in the compensated animals (bladder weights less than 5 g) and control animals. Blood flow to the mucosa followed the same pattern but without reaching significance. Blood flow returned to near baseline values within 5 minutes. Catheterization without drainage did not alter the blood flow. In contrast, drainage by puncture increased the blood flow significantly. Higher intravesical volumes increased the intravesical pressure slightly, but after opening the abdominal fascia, the intravesical pressure did not change with increasing volumes. CONCLUSIONS: Although the previously shown decreased blood flow to the bladder smooth muscle may be an etiologic factor in bladder contractile dysfunction secondary to partial outlet obstruction, the bladder does have the ability to increase the blood flow after drainage. This ability could be a compensatory and possibly protective mechanism after outlet obstruction. PMID- 11489731 TI - Partial ureteral obstruction dysregulates the renal renin-angiotensin system in the fetal sheep kidney. AB - OBJECTIVES: To investigate whether partial ureteral obstruction (PUO) in the fetus induces dysregulation of the renin-angiotensin system (RAS) and of transforming growth factor-beta 1 (TGF-beta1) and tissue inhibitors of metalloproteinase (TIMP1) expression. Previous studies have indicated that renal and urinary tract development depend on an intact renal RAS. Fetal urinary obstruction is distinct from postnatal obstruction. It has been suggested in postnatal animal studies that dysregulation of the RAS, and subsequent increased expression of TGF-beta1 and TIMP1, leads to changes in extracellular matrix composition. METHODS: Bilateral PUO was created in 4 fetal sheep. Seven animals (four obstructed and three controls) were killed at birth and their kidneys removed. Semiquantitative reverse transcriptase-polymerase chain reaction was used to quantify the levels of renin, angiotensinogen, angiotensin receptor type 1 (AT1 receptor), angiotensin receptor type 2 (AT2 receptor), TGF-beta1, and TIMP1. These messages were normalized to glyceraldehyde-3-phosphate dehydrogenase mRNA. RESULTS: All obstructed animals had moderate to severe hydronephrosis with enlarged kidneys (mean weight 22.0 g versus 9.4 g for the control animals; P <0.05). The increase in the levels of renin, angiotensinogen, AT1 receptor, TGF beta1, and TIMP1 mRNA was significant in the PUO group compared with the control group (P <0.05). AT2 receptor levels did not increase, but the AT1/AT2 mRNA ratio was significantly increased over normal (P <0.005). Also, a significant linear correlation was found between the increased renal weight and increased TGF-beta1 mRNA levels (P <0.005). CONCLUSIONS: Our findings suggest that fetal PUO can cause upregulation of the renal RAS and increased expression of TGF-beta1 and TIMP1, which may alter the balance between the generation and degradation of the extracellular matrix. The coordinate increases in renin, angiotensinogen, and AT1 receptor mRNA levels in chronic fetal PUO may represent a maladaptive response that contributes to interstitial fibrosis and prolonged vasoconstriction. RAS components and growth factors, particularly TGF-beta1, may be considered relevant targets in the prevention and treatment of congenital obstructive nephropathy. PMID- 11489732 TI - Advances in cytoarchitectonic mapping of the human cerebral cortex. AB - The classic cytoarchitectonic maps of the human cerebral cortex (e.g., that of Brodmann [1909]) considerably influenced the concept of localization of function. Although they suffer from drawbacks such as the highly observer-dependent definition of areal borders and the lack of information on intersubject variability of location and size of a cortical area in a spatial reference system, these maps often serve as anatomic references in functional imaging studies. Recent methodologic progress in computerized image analysis of histologic specimens and the development of warping techniques to compensate for intersubject variability of brain structure in the three-dimensional (3-D) space made it possible to overcome these drawbacks. The authors propose a new concept of cytoarchitectonic mapping, which is based on (1) a definition of areal borders using multivariate statistical analysis, (2) a quantitative analysis of similarity and dissimilarity in architecture between cortical areas, and (3) a probabilistic mapping of cytoarchitectonic areas in a 3-D reference system. The combined analysis of architectonic maps and functional imaging data in an identical reference space offers exciting perspectives in the analysis of structural-functional correlations in the living human brain and provides new insights into the architecture of the cerebral cortex. PMID- 11489733 TI - The motor cortex: anatomic substrates of function. AB - This article provides a firm foundation in the gross and microscopic anatomy of the motor cortex. It reviews the sites, Brodmann regions, normal function, and deficits associated with the primary motor cortex, supplementary motor area, presupplementary motor area, and the cingulate motor area in preparation for understanding the functional MR imaging studies that are reviewed in the later articles. PMID- 11489734 TI - Functional magnetic resonance imaging: factors modulating the cortical activation pattern of the motor system. AB - The use of functional MR imaging (fMRI), especially in a clinical setting, requires knowledge and control of the physiologic factors that can affect the hemodynamic response of the brain. In this article, the authors review the influence of elementary parameters of the motor system such as hemispheric dominance, rate, and force of action, and complexity of a paradigm in determining the cortical fMRI signal change. PMID- 11489736 TI - Clinical applications of functional magnetic resonance imaging. AB - This article addresses the application of clinical functional MR imaging to evaluate tumors, epilepsy, and questions of brain plasticity. PMID- 11489735 TI - Motor system: cortex, basal ganglia, and cerebellum. AB - In this article, the authors summarize results from imaging studies, analyzing the functional anatomy of motor sequence learning and timing. Emphasis is on the relationship between the cortical motor areas, the basal ganglia, and the cerebellum. PMID- 11489737 TI - Functional magnetic resonance imaging of human olfaction. AB - Olfaction is our basic sense phylogenetically and embryologically. Little is known, however, about how the human brain encodes the quality of odors, odor associated memories, and emotions. Olfactory information is projected from the olfactory bulb to the primary olfactory cortex, which is composed of the anterior olfactory nucleus, the olfactory tubercle, the piriform cortex, the amygdala, the periamygdaloid region, and the entorhinal cortex. From there, the primary olfactory cortex projects to secondary olfactory regions including the hippocampus, ventral striatum and pallidum, hypothalamus, thalamus, orbitofrontal cortex, agranular insular cortex, and cingulate gyrus. Functional MR studies using olfactory stimuli as paradigms show activation of many of these areas and can advance our understanding of odor perception in humans. PMID- 11489738 TI - Ocular motor system: anatomy and functional magnetic resonance imaging. AB - This article reviews current positron emission tomography and functional MR imaging (fMRI) studies of brain activation and indicates some of the insights they provide into the neuronal network mediating motion-perception and ocular motor control. The complexity of the network and the activation of the entire system seemingly independent of the specific ocular motor task performed makes functional interpretation extremely difficult. At the same time, however, the complexity illustrates the dilemma of associating a particular locus with one specific function. Caution is required to differentiate specific from nonspecific effects, to identify activations and deactivations, and to distinguish which of the systems has been activated inadvertently through poor stimulus control. The next step required for meaningful correlations of structure with function is for fMRI studies to test neurologic patients with distinct and separate lesions of the network. Such a comparison must evaluate the effect of the ocular motor deficit on the lesion site and the connected network in the acute stage and then following functional recovery caused by plasticity and neuronal repair. PMID- 11489739 TI - Vestibular system: anatomy and functional magnetic resonance imaging. AB - In this article, the authors present evidence that the multisensory parietoinsular cortex in humans is involved in perceiving both verticality and self-motion. This evidence is based on (a) lesional studies in patients who have acute unilateral infarctions and (b) positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) evidence of cerebral activation by use of vestibular caloric stimulation, galvanic stimulation, and optokinetic stimulation. PMID- 11489740 TI - Auditory system: functional magnetic resonance imaging. AB - Functional MR imaging (fMRI) is being used increasingly to explore the human central auditory system. The considerable background noise produced by echo planar imaging (EPI) and other fMRI sequences, however, interferes in an unpredictable way with the experimental stimuli. Several approaches exist to overcome this problem. Each has its advantages and disadvantages. These different approaches allow researchers to tailor the experimental designs to specific research questions. Recent studies have yielded significant information about human auditory function. Compared with other sensory systems such as the visual system, the auditory database still is relatively small. It is expected that novel methodologic approaches will stimulate scientific exploration of auditory processing and eventually lead to clinically meaningful applications of auditory fMRI. PMID- 11489741 TI - Functional magnetic resonance imaging of the auditory cortex as a diagnostic tool in cochlear implant candidates. AB - Current use of functional MR imaging (fMRI) on cochlea implant candidates is reviewed in this article. Also included are problems of using promontory testing (PT) inside the MR scanner and the results of the latest studies and illustrative cases of fMRI of the auditory cortex in deaf patients using PT for stimulation. PMID- 11489742 TI - Anatomic substrates of language: emphasizing speech. AB - This article reviews in detail the gross anatomy of the auditory and language areas, the present concepts of unimodal and multimodal association cortices, the classic model of language, problems with the classic model, and newer approaches to conceptualizing the structural interconnections that subserve audition and language. PMID- 11489743 TI - Approach to functional magnetic resonance imaging of language based on models of language organization. AB - Functional MR imaging (fMRI) has been a useful tool in the evaluation of language both in normal individuals and patient populations. The purpose of this article is to use various models of language as a framework to review fMRI studies. Specifically, fMRI language studies are subdivided into the following categories: word generation or fluency, passive listening, orthography, phonology, semantics, and syntax. PMID- 11489744 TI - Determination of cerebral hemisphere language dominance with functional magnetic resonance imaging. AB - Functional MR imaging (fMRI) is useful for the determination of cerebral hemisphere language dominance, but it can be misleading if the imaging studies are not performed properly. The theoretic and technical problems involved in this use of fMRI are discussed, including behavioral paradigms, data analysis methods, subject handedness, and gender. PMID- 11489745 TI - Functional magnetic resonance imaging research in psychiatry. AB - Advanced functional imaging techniques now make it possible to study in vivo the relationship between altered cerebral activation patterns and the psychopathologic and cognitive features of psychiatric disorders. Functional MR imaging (fMRI) offers new approaches to research questions in psychiatry that could not have been addressed earlier by positron emission tomography and single photon emission CT. Basic features of the biology of psychiatric disorders now can be elucidated by means of fMRI. Study designs involving fMRI capitalize on the technique's enhanced spatial and temporal resolution, noninvasive nature, and absence of radiation exposure. PMID- 11489746 TI - Pharmacologic modulation of human cerebral activity: contribution of functional neuroimaging. AB - This article highlights the following: (1) How the tools of functional neuroimaging analyze the effect of drug therapy on the central nervous system; (2) how the interactions between the drugs and the cortical neuronal networks have illustrated new aspects of the concepts of cerebral neuroplasticity; and (3) how functional neuroimaging has proved to be a valuable tool in understanding the mechanisms that govern the therapeutic activity of experimental drugs. PMID- 11489747 TI - Alcohol consumption and lung cancer: a review of the epidemiologic evidence. AB - A role for alcohol consumption in lung cancer etiology has been suggested in some studies, but this possible relationship has been often regarded with skepticism, with any indication of an association being attributed to confounding by cigarette smoking. The purpose of this work was to review the epidemiological evidence for an association of alcohol and lung cancer and to identify gaps in that research. The studies reviewed here provide some indication that alcohol and particularly beer intake may increase lung cancer risk after controlling for cigarette smoking. Although the evidence is not conclusive, it warrants additional consideration of alcohol as a risk factor in lung cancer etiology, independent of cigarette smoking. Recommendations for future studies are provided. PMID- 11489748 TI - The impact of smoking status, disease stage, and index tumor site on second primary tumor incidence and tumor recurrence in the head and neck retinoid chemoprevention trial. AB - Second primary tumors (SPTs) develop at an annual rate of 3-7% in patients with head and neck squamous cell cancer (HNSCC). In a previous Phase III study, we observed that high doses of 13-cis-retinoic acid reduced the SPT rate in this disease. In 1991, we launched an intergroup, placebo-controlled, double-blind study to evaluate the efficacy of low-dose 13-cis-retinoic acid in the prevention of SPTs in patients with stage I or II squamous cell carcinoma of the larynx, oral cavity, or pharynx who had been previously successfully treated with surgery, radiotherapy, or both, and whose diagnoses had been established within 36 months of study entry. As of September 16, 1999, the Retinoid Head and Neck Second Primary (HNSP) Trial had completed accrual with 1384 registered patients and 1191 patients randomized and eligible. All of the patients were followed for survival, SPT development, and index cancer recurrence. Smoking status was assessed at study entry and during study. Smoking cessation was confirmed biochemically by measurement of serum cotinine levels. The annual rate of SPT development was analyzed in terms of smoking status and tumor stage. As of May 1, 2000, SPTs have developed in 172 patients. Of these, 121 (70.3%) were tobacco related SPTs, including 113 in the aerodigestive tract (57 lung SPTs, 50 HNSCC SPTs, and 6 esophageal SPTs) and 8 bladder SPTs. The remaining 51 cases included 23 prostate adenocarcinomas, 8 gastrointestinal malignancies, 6 breast cancers, 3 melanomas, and 11 other cancers. The annual rate of SPT development observed in our study has been 5.1%. SPT development related to smoking status was marginally significant (active versus never, 5.7% versus 3.5%; P = 0.053). Significantly different smoking-related SPT development rates were observed in current, former, and never smokers (annual rate = 4.2%, 3.2%, and 1.9%, respectively, overall P = 0.034; current versus never smokers, P = 0.018). Stage II HNSCC had a higher overall annual rate of SPT development (6.4%) than did stage I disease (4.3%; P = 0.004). When evaluating the development of smoking-related SPTs, stage was also highly significant (4.8% for stage II versus 2.7% for stage I; P = 0.001). Smoking-related SPT incidence was significant for site as well (larynx versus oral cavity, P = 0.015; larynx versus pharynx, P = 0.011). Primary tumors recurred at an annual rate of 2.8% in a total of 97 patients. The rate of recurrence was higher in patients with stage II disease (4.1% versus 2.2%, P = 0.004) as well as oral cavity site when compared with larynx (P = 0.002). This is the first large-scale prospective chemoprevention study evaluating smoking status and its impact on SPT development and recurrence rate in HNSCC. The results indicate significantly higher SPT rates in active smokers versus never smokers and significantly higher smoking-related SPT rates in active smokers versus never smokers, with intermediate rates for former smokers. PMID- 11489749 TI - Protein adducts of 1,4-benzoquinone and benzene oxide among smokers and nonsmokers exposed to benzene in China. AB - Hemoglobin (Hb) and albumin (Alb) adducts of the benzene metabolites benzene oxide (BO) and 1,4-benzoquinone (1,4-BQ) were analyzed by gas chromatography-mass spectrometry in 43 exposed workers and 44 unexposed controls from Shanghai, China, as part of a larger cross-sectional study of benzene biomarkers. When subjects were divided into controls (n = 44) and workers exposed to 31 ppm (n = 22) of benzene, median 1,4-BQ-Alb adducts were 2110, 5850, and 13,800 pmol/g Alb, respectively (correlation with exposure: Spearman r = 0.762; P < 0.0001); median BO-Alb adducts were 106, 417, and 2400 pmol/g Alb, respectively (Spearman r = 0.877; P < 0.0001); and median BO-Hb adducts were 37.1, 50.5, and 136 pmol/g Hb, respectively (Spearman r = 0.757; P < 0.0001). To our knowledge, this is the first observation that adducts of 1,4-BQ are significantly correlated with benzene exposure. When compared on an individual basis, Alb adducts of 1,4-BQ and BO and Hb adducts of BO were highly correlated with each other and with urinary phenol and hydroquinone (P < 0.0001 for all of the comparisons). Although detectable in the assays, Hb adducts of 1,4-BQ and both Hb and Alb adducts of 1,2-BQ produced erratic results and are not reported. Interestingly, cigarette smoking increased Alb adducts of 1,4-BQ but not of BO, suggesting that benzene from cigarette smoke was not the primary contributor to the 1,4-BQ adducts. PMID- 11489750 TI - Human lung microsomal cytochrome P4501A1 (CYP1A1) activities: impact of smoking status and CYP1A1, aryl hydrocarbon receptor, and glutathione S-transferase M1 genetic polymorphisms. AB - There are numerous conflicting epidemiological studies addressing correlations between cytochrome P450 1A1 (CYP1A1) genetic polymorphisms and lung cancer susceptibility, with associations plausibly linked to alterations in carcinogen bioactivation. Similarly, correlations between aryl hydrocarbon receptor gene (AHR) codon 554 genotype and CYP1A1 inducibility are controversial. The objective of this study was to determine whether smoking status, and CYP1A1, AHR, and glutathione S-transferase M1 gene (GSTM1) polymorphisms correlate with altered CYP1A1 activities. Lung microsomal CYP1A1-catalyzed 7-ethoxyresorufin O dealkylation (EROD) activities were much higher in tissues from current smokers (n = 46) than in those from non-/former smokers (n = 24; 12.11 +/- 13.46 and 0.77 +/- 1.74 pmol/min/mg protein, respectively, mean +/- SD; P < 0.05). However, EROD activities in lung microsomes from current smokers CYP1A1*1/1 (n = 33) and heterozygous MspI variant CYP1A1*1/2A (n = 10) were not significantly different (12.23 +/- 13.48 and 8.23 +/- 9.76 pmol/min/mg protein, respectively, P > 0.05). Three current smokers were heterozygous variant CYP1A1*1/2B (possessing both *2A and *2C alleles), and exhibited activities similar to individuals CYP1A*1/1. One current smoker was heterozygous variant CYP1A1*4 and exhibited activities comparable with individuals CYP1A1*1/1 at that locus. EROD activities in microsomes from current smokers AHR(554)Arg/Arg (n = 41) and heterozygous variant AHR(554)Arg/Lys (n = 5) were not significantly different (12.13 +/- 13.56 and 12.01 +/- 14.23 pmol/min/mg protein, respectively; P > 0.05). Furthermore, microsomal EROD activities from current smokers with the GSTM1-null genotype (n = 28) were not significantly different from those (n = 18) carrying at least one copy of GSTM1 (12.61 +/- 14.24 and 11.34 +/- 12.53 pmol/min/mg protein, respectively; P > 0.05). Additionally, when genotypic combinations of CYP1A1, AHR, and GSTM1 were assessed, there were no significant effects on EROD activity. On the basis of microsomal enzyme activities from heterozygotes, CYP1A1*1/2A, CYP1A1*1/2B, CYP1A1*1/4, and AHR(554) Arg/Lys variants do not appear to significantly affect CYP1A1 activities in human lung, and we observed no association between CYP1A1 activity and the GSTM1-null polymorphism. PMID- 11489751 TI - Activities and exposures during leisure and prostate cancer risk. AB - The objective of this study was to identify activities and exposures during leisure that might be associated with the development of prostate cancer. We analyzed data derived from a population-based case-control study that was carried out in Montreal between 1979 and 1985. Men (>4000) were interviewed, including cases of prostate cancer, other cancers, and population controls. The present analysis was restricted to the subset, aged 45-70 years, who underwent face-to face interviews in which aspects of activities and exposures during leisure were ascertained. There were 400 incident cases of prostate cancer and 476 population controls. We calculated odds ratios (OR) for prostate cancer, adjusted for age, ethnic origin, respondent status, family income, body mass index, cigarette smoking, and alcohol consumption. Home or furniture maintenance was associated with an increased risk [OR, 1.4; 95% confidence interval (CI), 1.0-1.9], as was painting, stripping, or varnishing furniture (OR, 2.1; 95% CI, 0.7-6.7). Exposure during leisure to metal dust was associated with prostate cancer (OR, 3.2; 95% CI, 1.0-9.9), as was exposure to lubricating oils or greases (OR, 2.2; 95% CI, 1.2-3.7) and exposure to pesticides or garden sprays (OR, 2.3; 95% CI, 1.3-4.2). These findings are consistent with results derived from studies of occupational exposures. PMID- 11489753 TI - Vitamin D receptor polymorphism and the risk of colorectal adenomas: evidence of interaction with dietary vitamin D and calcium. AB - Laboratory studies and epidemiological investigations suggest that vitamin D plays a role in the etiology of colorectal adenomas, possibly through a mechanism mediated by the vitamin D receptor (VDR). We conducted a clinic-based case control study to examine the association between VDR polymorphisms and colorectal adenomas. We selectively identified a random subset of 393 cases of colorectal adenomas and 406 colonoscopy-negative controls from a clinic-based case-control study conducted in the metropolitan Minneapolis/St. Paul area during 1991-1994. A self-administered questionnaire was used to collect data on dietary and supplement intake of vitamin D and calcium, as well as on demographics, physical activity, medical information, lifestyle factors, reproductive history, and anthropometry. DNA was extracted from whole blood and assayed for the BsmI VDR polymorphism using an ABI 7700 TaqMan assay. Adjusted odds ratios (OR) and 95% confidence intervals (CIs) were evaluated using logistic regression. Compared with the bb genotype (33% of controls), neither the Bb (48.8% of controls) nor the BB (18.2% of controls) genotypes was strongly associated with risk of colorectal adenomas (OR = 0.86, CI = 0.63-1.19 and OR = 0.77, CI = 0.50-1.18, respectively). However, those with the lowest tertile of vitamin D intake and the BB genotype had a lower risk of colorectal adenoma (OR = 0.24, CI = 0.08-0.76) than those with the highest tertile of intake and the bb genotype. Similarly, those with the lowest tertile of calcium intake and the BB genotype had a reduced risk of colorectal adenoma (OR = 0.34, CI = 0.11-1.06). Although it has generally been shown that higher calcium and vitamin D intake are associated with a modestly reduced risk of colorectal neoplasia, our data suggest that those with the BB BsmI VDR genotype may be at reduced risk of colorectal adenoma in the presence of lower calcium and vitamin D intake. PMID- 11489752 TI - Phase II randomized clinical trial of lycopene supplementation before radical prostatectomy. AB - An inverse association has been observed between dietary intake of lycopene and the risk of prostate cancer. We investigated the effects of lycopene supplementation in patients with prostate cancer. Twenty-six men with newly diagnosed, clinically localized (14 T(1) and 12 T(2)) prostate cancer were randomly assigned to receive 15 mg of lycopene (n = 15) twice daily or no supplementation (n = 11) for 3 weeks before radical prostatectomy. Biomarkers of differentiation and apoptosis were assessed by Western blot analysis on benign and malignant parts of the prostate gland. Prostatectomy specimens were entirely embedded, step-sectioned, and evaluated for pathological stage, Gleason score, volume of cancer, and extent of high-grade prostatic intraepithelial neoplasia. Plasma levels of lycopene, insulin-like growth factor-1 (IGF-1), IGF binding protein-3, and prostate-specific antigen were measured at baseline and after 3 weeks of supplementation or observation. Eleven (73%) subjects in the intervention group and two (18%) subjects in the control group had no involvement of surgical margins and/or extra-prostatic tissues with cancer (P = 0.02). Twelve (84%) subjects in the lycopene group and five (45%) subjects in the control group had tumors <4 ml in size (P = 0.22). Diffuse involvement of the prostate by high grade prostatic intraepithelial neoplasia was present in 10 (67%) subjects in the intervention group and in 11 (100%) subjects in the control group (P = 0.05). Plasma prostate-specific antigen levels decreased by 18% in the intervention group, whereas they increased by 14% in the control group (P = 0.25). Expression of connexin 43 in cancerous prostate tissue was 0.63 +/- 0.19 absorbance in the lycopene group compared with 0.25 +/- 0.08 in the control group (P = 0.13). Expression of bcl-2 and bax did not differ significantly between the two study groups. IGF-1 levels decreased in both groups (P = 0.0002 and P = 0.0003, respectively). The results suggest that lycopene supplementation may decrease the growth of prostate cancer. However, no firm conclusions can be drawn at this time because of the small sample size. PMID- 11489754 TI - Epoxide hydrolase Tyr113His polymorphism is associated with elevated risk of colorectal polyps in the presence of smoking and high meat intake. AB - Microsomal epoxide hydrolase (mEH) metabolizes polycyclic aromatic hydrocarbons, carcinogens found in cigarette smoke and cooked meat. Polymorphisms in exon 3 and exon 4 of the mEH gene have been found to alter mEH activity. We investigated the association between these polymorphisms and colorectal polyps within the Minnesota Cancer Prevention Research Unit case-control study. Cases were diagnosed with colonoscopically confirmed adenomas (n = 530) or hyperplastic polyps (n = 202); controls (n = 649) were polyp-free at colonoscopy. Smoking history and meat consumption were obtained from self-administered questionnaires before colonoscopy. mEH genotypes were determined by PCR/RFLP or oligonucleotide ligation assay. The overall risks associated with exon 3 or exon 4 polymorphisms for both adenomas and hyperplastic polyps were not statistically different from 1.0. Compared with exon 3 Tyr/Tyr, 0 pack-years, risk was highest among those with the exon 3 His/His genotype and >25 pack-years of smoking [adenoma, odds ratio (OR) = 4.9 (1.9-12.8); hyperplastic, OR = 7.7 (2.5-24.0)]. Risks were not elevated among exon 4 homozygous variants, even in the presence of heavy smoking. Fried, baked, or broiled meat intake of > or =two servings/week (high) compared with < or =one serving/week was associated with a 2-fold increase in risk of adenoma. The highest risks were seen for those with the exon 3 His/His genotype and high cooked meat intake [OR = 3.3 (1.4-7.9); reference group: Tyr/Tyr, < or = 1 serving/week). Although mEH polymorphisms are not associated with an increased risk of colorectal polyps overall, genotypes that produce a slow phenotype appear to be associated with an increased risk in the presence of smoking and high intakes of cooked meat. PMID- 11489755 TI - Cancer risk assessment for the environmental mutagen and carcinogen crotonaldehyde on the basis of TD(50) and comparison with 1,N(2) propanodeoxyguanosine adduct levels. AB - Humans are ubiquitously exposed to crotonaldehyde to a strongly varying extent, in particular, via food and alcoholic beverages. Like other alpha,beta unsaturated carbonyl compounds, crotonaldehyde forms 1,N(2)-propanodeoxyguanosine adducts and is genotoxic, mutagenic, and carcinogenic. This study was designed to perform a cancer risk assessment on the basis of TD(50), which was available from a long-term cancer study with F-344 rats (F. L. Chung et al., Cancer Res., 46: 1285-1289, 1986), and the estimated daily intake via food and beverages. A relatively high cancer risk of 0.1-1 cancer incidence/10(3) humans was extrapolated on the basis of the TD(50) from the cancer study of Chung et al. for the estimated dietary intake and drinking wine. To compare the 1,N(2) propanodeoxyguanosine DNA adduct levels of crotonaldehyde with the assessed cancer risk, we synthesized adduct standards and developed a (32)P-postlabeling method for DNA adducts of crotonaldehyde providing a detection limit of 3 adducts/10(9) nucleotides. Repeated gavages of 10 and 1 mg/kg were given to simulate the steady-state situation of the animal cancer study of Chung et al. and to estimate the adduct levels after intake of crotonaldehyde via food. The estimated adduct levels at these crotonaldehyde intakes were in the range of 3 adducts/10(9) nucleotides. The adducts persisted to a certain extent. The persistence is important for considering the steady-state situation after permanent intakes of crotonaldehyde via food. However, the adducts are repaired to some extent; 2 weeks after the last of repeated gavages, only 19% of the initial amount measured directly after the last gavage is left. According to our results, a steady-state concentration in the range of 3 adducts/10(9) nucleotides is responsible for the induction of cancer in the study of Chung et al., in the case that cancer from crotonaldehyde depends exclusively on the 1,N(2) propanodeoxyguanosine adducts considered here. No propanodeoxyguanosine adducts of crotonaldehyde were found in the DNA of untreated animals in our studies. PMID- 11489756 TI - Biomarker modulation in a nonhuman rhesus primate model for ovarian cancer chemoprevention. AB - OBJECTIVE: The objective of this study was to explore whether a nonhuman primate model could be developed to test drugs for the prevention of ovarian cancer. METHODS: Nineteen adult female Rhesus macaques were given fenretinide (4HPR), oral contraceptives (OCP), the combination (4HPR + OCP), or no medication for 3 months. Exploratory laparotomy was done pre- and postdrug to assess intermediary biomarkers of neoplastic phenotype, proliferation, response pathways, and growth regulatory and metabolic markers. Fluorescence emission spectra were plotted for each group pre- and postdrug and means were overlaid on these plots and normalized. Fluorescence intensities were compared using the 2-tailed Student t test, (P = 0.1-0.01). RESULTS: All monkeys tolerated drugs and surgeries without difficulty. Histochemical markers showed no significant trend. However, fluorescence spectroscopy showed increased intensity at 450 nm excitation, 550 nm emission correlating with increased FAD presence. The 4HPR group (P = 0.01) showed higher intensity than the OCP group (P = 0.05-0.07) when compared with the controls. Decreased emission was seen at 350 nm excitation, 450 nm emission correlating with decreased NAD(P)H presence. The OCP group showed the largest change (P < 0.01), and the control group showed the smallest change. CONCLUSIONS: The nonhuman primate is an excellent model to test drug effect on the ovarian surface epithelium and merits additional study. Fluorescence spectroscopy was the most sensitive marker for drug activity and the apparent increase in NAD and FAD in the 4HPR group is consistent with the effect of 4HPR observed in cell culture. The differences between the OCP and the 4HPR groups suggest a different mechanism of activity of these drugs. PMID- 11489757 TI - Communicating breast cancer risks to women using different formats. AB - Using a pre-post test design with a baseline, laboratory, and a 6-month follow up, we communicated women's objective breast cancer risks, based on the Gail Model, using two formats: (a) range of risks (e.g., risk of breast cancer can be as low as 1% and as high as 5%); and (b) as a point estimate (e.g., your risk of breast cancer is 3%). We examined how these presentations individually and jointly affected women's perceived lifetime breast cancer risks. Overall, providing risk estimates either as a range of risks or as a point estimate lowered women's perceived lifetime risks compared with women who did not get information presented this way shortly after receipt of this information relative to baseline. At the 6-month follow-up, perceptions of lifetime risks generally returned to their baseline values. Overall, women viewed their risk feedback, whether presented as a point estimate or as a range of risks, as equally credible, trustworthy, accurate, and personally relevant. These results suggest that women evaluate risk feedback containing either point estimates or range of risks as equally acceptable. Both formats lead to short-term reductions in perceived risk (i.e., greater accuracy). PMID- 11489758 TI - Alcoholism and risk for cancer of the cervix uteri, vagina, and vulva. AB - We conducted a population-based cohort study to analyze the risk of developing cancers of the female genitals among 36,856 patients with a hospital discharge diagnosis of alcoholism (ICD-7: 307, 322; ICD-8: 291, 303; ICD-9: 291, 303, 305A) in Sweden between 1965 and 1995. The follow-up was done by linkages of national registries. Standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) were computed based on nationwide specific cancer rates. The first year of follow-up was excluded from all analyses to minimize the impact of selection bias. We found that alcoholic women had excess risks for in situ cervical cancer (SIR, 1.7; 95% CI, 1.6-1.9), for invasive cervical cancer (SIR, 2.9; 95% CI, 2.4 3.5), and for cancer of the vagina (SIR, 4.6; 95% CI, 2.2-8.5) but not for cancer of the vulva (SIR, 1.0; 95% CI, 0.4-2.0). The fact that alcoholics had an excess risk also for the in situ cancer suggests that the observed excess in invasive cervical cancer may not only be attributable to less use of Pap smear screening among them. The alcoholic women may be at higher risk for the progression from human papillomavirus infection to a malignant lesion for lifestyle-related reasons (promiscuity, smoking, use of contraceptive hormones, and dietary deficiencies). We conclude that alcoholic women are at high risk for in situ and invasive cervical cancer and for cancer of the vagina. PMID- 11489759 TI - Regular use of analgesic drugs and ovarian cancer risk. AB - Analgesics have been shown to reduce risk for colorectal cancer. Results from three recent reports (D. W. Cramer et al., Lancet, 351: 104-107, 1998; C. Rodriguez et. al., Lancet, 352: 1354-1355, 1998; L. Rosenberg et al., Cancer Epidemiol. Biomark. Prev., 9: 933-937, 2000) suggest that these drugs might be associated with decreased risk for ovarian cancer. In this hospital-based case control study, we compared 547 patients with ovarian cancer to 1094 age-matched patients with nonneoplastic conditions. All of the participants received treatment at the Roswell Park Cancer Institute between 1982 and 1998 and completed a comprehensive epidemiological questionnaire that included information on demographics, life-style factors, and reproductive characteristics as well as frequency and duration of aspirin and acetaminophen use. Women who reported that they had used one or more of these agents at least once a week for at least 6 months were classified as analgesic users. Logistic regression was used to compute crude and adjusted odds ratios (ORs) with 95% confidence intervals (CIs). Aspirin users were not at reduced risk of ovarian cancer compared with nonusers (adjusted OR, 1.00; CI, 0.73-1.39). There was also no evidence of a decrease in risk as a function of greater frequency of use or prolonged duration of use. Regular acetaminophen use was associated with a reduced risk (adjusted OR, 0.56; 95% CI, 0.34-0.86), and risk reductions were observed for women with the greatest frequency of use (adjusted OR, 0.32; 95% CI, 0.09-1.08) and longest duration of use (adjusted OR, 0.51; 95% CI, 0.27-0.97). These data suggest that regular use of acetaminophen, but not aspirin, may be associated with lower risk of ovarian cancer. PMID- 11489760 TI - Exposure to environmental tobacco smoke is associated with lower plasma beta carotene levels among nonsmoking women married to a smoker. AB - We evaluated the association between exposure to environmental tobacco smoke (ETS) from husbands who smoke and plasma levels of antioxidant vitamins among nonsmoking women. A total of 1249 women from four areas in Italy answered a self administered questionnaire, reported their diets on a food frequency questionnaire, had a medical examination, and gave their blood for alpha and beta carotene, retinol, L-ascorbic acid, alpha-tocopherol, and lycopene determinations. Urinary cotinine was used to evaluate the level of recent exposure to ETS. After adjusting for study center, age and education, we found no association between ETS exposure and daily nutrient intake of beta-carotene, retinol, L-ascorbic acid, and alpha-tocopherol. However, we found an inverse dose response relationship between intensity of current husband's smoke and concentrations of plasma beta-carotene and L-ascorbic acid. The associations remained even after controlling for daily beta-carotene and vitamin C intake and for other potential confounders (vitamin supplementation, alcohol consumption, and body mass index). Moreover, when urinary cotinine was considered as the exposure variable, a significant inverse association with plasma beta-carotene was found. The findings may be of interest to explain the biological mechanism that link ETS exposure with lung cancer and ischemic heart diseases. PMID- 11489761 TI - No association between the XPD (Lys751G1n) polymorphism or the XRCC3 (Thr241Met) polymorphism and lung cancer risk. PMID- 11489762 TI - No association between fat and fatty acids intake and risk of colorectal cancer. PMID- 11489764 TI - Importance of TIMI 3 flow. PMID- 11489765 TI - Troponisms, necrosettes, enzyme leaks, creatinine phosphokinase bumps, and infarctlets: what's behind this new lexicon and what does it add? PMID- 11489766 TI - Admission troponin T level predicts clinical outcomes, TIMI flow, and myocardial tissue perfusion after primary percutaneous intervention for acute ST-segment elevation myocardial infarction. AB - BACKGROUND: In ST-segment elevation myocardial infarction, a troponin T >/=0.1 microg/L on admission indicates poorer prognosis despite early reperfusion. To evaluate the underlying reason, we studied the value of cardiac troponin T (cTnT) for prediction of outcomes, epicardial blood flow, and myocardial reperfusion after primary percutaneous intervention. METHODS AND RESULTS: Patients (n=140) admitted within 12 hours after onset of symptoms were stratified by admission cTnT. Epicardial and myocardial reperfusion were graded by the TIMI score and by measurement of relative increases of myoglobin, cTnT, and creatine kinase (CK)-MB 60 minutes after recanalization, respectively. cTnT was positive in 64 patients (45.7%) and was associated with longer median time intervals to admission (5.5 versus 3.5 hours, P<0.001) and higher mortality rates after 30 days (12.5% versus 3.9%, P=0.06) and 9 months (14% versus 3.9%, P=0.005). cTnT independently predicted a 3.2-fold risk for incomplete epicardial reperfusion (P=0.03). In addition, cTnT >/=0.1 microg/L was associated with more severely impaired myocardial perfusion despite normal epicardial flow, as indicated by lower 60 minute ratios of myoglobin (2.6 versus 7.6, P=0.007), cTnT (6.6 versus 29.2, P<0.001), and CK-MB (3.5 versus 21.4, P=0.002) and a tendency for less resolution of ST-segment elevations (54% versus 60%, P=0.08). CONCLUSIONS: cTnT predicts poorer clinical outcomes, lower rates of postprocedural TIMI 3 flow, and more severely compromised myocardial perfusion despite normal epicardial flow. Thus, a cTnT-positive patient may require more aggressive adjunctive therapy when treated by percutaneous coronary intervention. The impact of preexisting or evolving microvascular dysfunction and the effect of therapies that target myocardial perfusion require further prospective evaluation. PMID- 11489767 TI - Normal flow (TIMI-3) before mechanical reperfusion therapy is an independent determinant of survival in acute myocardial infarction: analysis from the primary angioplasty in myocardial infarction trials. AB - BACKGROUND: Whereas survival after lytic therapy for myocardial infarction is strongly dependent on early administration, it is unknown whether the otherwise excellent outcomes in patients undergoing primary PTCA for acute myocardial infarction, in whom TIMI-3 flow rates of >90% may be achieved, can be further improved by early reperfusion. METHODS AND RESULTS: Among 2507 patients enrolled in 4 PAMI trials undergoing primary PTCA, spontaneous reperfusion (TIMI-3 flow) was present in 16% at initial angiography. Compared with patients without TIMI-3 flow, those with TIMI-3 flow before PTCA had greater left ventricular ejection fraction (57+/-10% versus 53+/-11%, P=0.003) and were less likely to present in heart failure (7.0% versus 11.6%, P=0.009). Patients with initial TIMI-3 flow had significantly lower in-hospital rates of mortality, new-onset heart failure, and hypotension and had a shorter hospital stay. Cumulative 6-month mortality was 0.5% in patients with initial TIMI-3 flow, 2.8% with TIMI-2 flow, and 4.4% with initial TIMI-0/1 flow (P=0.009). By multivariate analysis, TIMI-3 flow before PTCA was an independent determinant of survival (odds ratio 2.1, P=0.04), even when corrected for by postprocedural TIMI-3 flow. CONCLUSIONS: Patients undergoing primary PTCA in whom TIMI-3 flow is present before angioplasty present with greater clinical and angiographic evidence of myocardial salvage, are less likely to develop complications related to left ventricular failure, and have improved early and late survival. These data warrant prospective randomized trials of pharmacological strategies to promote early reperfusion before definitive mechanical intervention in acute myocardial infarction. PMID- 11489768 TI - Differential impact on survival of electrocardiographic Q-wave versus enzymatic myocardial infarction after percutaneous intervention: a device-specific analysis of 7147 patients. AB - BACKGROUND: The relative prognostic importance of ECG myocardial infarction (MI) after intervention compared with varying degrees of enzymatic elevation has not been characterized, and the device-specific implications of periprocedural MI are also unknown. METHODS AND RESULTS: Serial creatine phosphokinase (CPK)-MB levels were determined after elective percutaneous intervention of 12 098 lesions in 7147 consecutive patients at a tertiary referral center. Procedural, in-hospital, and follow-up data were collected by independent research nurses, and clinical and ECG events were adjudicated by a separate committee. Stents were implanted in 50.6% of lesions, atheroablation was performed in 54.8%, and PTCA alone was performed in 9.8%. The peak periprocedural CPK-MB level was >3x the upper limit of normal (ULN) in 17.9% of patients, and Q-wave MI developed in 0.6%. By multivariate analysis, the periprocedural development of new Q waves was the most powerful independent determinant of death (2-year mortality rate, 38.3%; hazard ratio, 9.9; P<0.0001). Non-Q-wave MI with CPK-MB >8x ULN was also a strong predictor of death (2-year mortality rate, 16.3%; hazard ratio, 2.2; P<0.0001); survival was unaffected by lesser degrees of CPK-MB elevation. Though CPK-MB elevation was more common after atheroablation and stenting than PTCA, the rates of Q-wave MI and survival were device-independent. CONCLUSIONS: Myonecrosis after percutaneous intervention is common in a high-risk referral population dominated by atheroablation and stent use. Large periprocedural infarctions (signified by new Q waves and CPK-MB >8xULN) are powerful determinants of death, whereas lesser degrees of CPK-MB release and specific device use do not adversely affect survival. PMID- 11489769 TI - Randomized comparison of enoxaparin, a low-molecular-weight heparin, with unfractionated heparin adjunctive to recombinant tissue plasminogen activator thrombolysis and aspirin: second trial of Heparin and Aspirin Reperfusion Therapy (HART II). AB - BACKGROUND: Adjunctive unfractionated heparin (UFH) during thrombolytic therapy for acute myocardial infarction (AMI) promotes the speed and magnitude of coronary artery recanalization and reduces reocclusion. Low-molecular-weight heparins offer practical and potential pharmacological advantages over UFH in multiple applications but have not been systematically studied as adjuncts to fibrinolysis in AMI. METHODS AND RESULTS: Four hundred patients undergoing reperfusion therapy with an accelerated recombinant tissue plasminogen activator regimen and aspirin for AMI were randomly assigned to receive adjunctive therapy for at least 3 days with either enoxaparin or UFH. The study was designed to show noninferiority of enoxaparin versus UFH with regard to infarct-related artery patency. Ninety minutes after starting therapy, patency rates (thrombolysis in myocardial infarction [TIMI] flow grade 2 or 3) were 80.1% and 75.1% in the enoxaparin and UFH groups, respectively. Reocclusion at 5 to 7 days from TIMI grade 2 or 3 to TIMI 0 or 1 flow and TIMI grade 3 to TIMI 0 or 1 flow, respectively, occurred in 5.9% and 3.1% of the enoxaparin group versus 9.8% and 9.1% in the UFH group. Adverse events occurred with similar frequency in both treatment groups. CONCLUSIONS: Enoxaparin was at least as effective as UFH as an adjunct to thrombolysis, with a trend toward higher recanalization rates and less reocclusion at 5 to 7 days. PMID- 11489770 TI - Lumen loss in transplant coronary artery disease is a biphasic process involving early intimal thickening and late constrictive remodeling: results from a 5-year serial intravascular ultrasound study. AB - BACKGROUND: Coronary artery disease is the major cause of late cardiac allograft failure. However, few data exist regarding the natural history of changes in intimal and external elastic membrane (EEM) areas after heart transplantation. METHODS AND RESULTS: In 38 transplant recipients, serial intravascular ultrasound examinations were performed 3.7+/-2.2 weeks after transplantation and annually thereafter for 5 years. In 59 coronary arteries, we compared 135 matched segments among serial studies. In each segment, intravascular ultrasound images were digitized at 1-mm intervals, and mean values of EEM and lumen and intimal areas were analyzed. In the first year after transplantation, the intimal area increased significantly from 1.8+/-1.6 to 3.0+/-2.1 mm(2) (P<0.001). Subsequently, the annual increase in intimal area decreased. EEM area did not change during the first year; however, between years 1 and 3, significant expansion of EEM area occurred (15.4+/-4.6 to 17.2+/-5.4 mm(2), P<0.001). Thereafter, EEM area decreased significantly from 17.2+/-5.4 mm(2) (year 3) to 15.1+/-4.9 mm(2) (year 5, P=0.01). Different mechanisms of lumen loss were observed during 2 phases after transplantation: early lumen loss primarily caused by intimal thickening and late lumen loss caused by EEM area constriction. CONCLUSIONS: This serial ultrasound study revealed that most of the intimal thickening occurred during the first year after heart transplantation. Changes in the EEM area showed a biphasic response, consisting of early expansion and late constriction. Thus, different mechanisms of lumen loss were observed during the early and late phases after transplantation. PMID- 11489771 TI - Reduced myocardial sarcoplasmic reticulum Ca(2+)-ATPase mRNA expression and biphasic force-frequency relations in patients with hypertrophic cardiomyopathy. AB - BACKGROUND: The relationship between left ventricular (LV) contractile functional reserve and gene expression of Ca(2+)-handling proteins in patients with hypertrophic cardiomyopathy (HCM) remains to be clarified. METHODS AND RESULTS: We calculated the maximum first derivative of LV pressure (LV dP/dt(max)) and the LV pressure half-time (T(1/2)) during pacing in 14 patients with nonobstructive HCM (LV ejection fraction >55%) and 7 control subjects. Endomyocardial tissue was obtained, and mRNA levels of sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2), ryanodine receptor-2, phospholamban, calsequestrin, and Na(+)/Ca(2+) exchanger were quantified by use of a real-time quantitative reverse transcription polymerase chain reaction method. Group A consisted of 7 HCM patients who showed a progressive rise in the LV dP/dt(max) with increased heart rate. Group B consisted of 7 HCM patients in whom the heart rate-LV dP/dt(max) relation was biphasic at physiological pacing rates. Both the mean maximal wall thickness and the LV hypertrophy score in group B were greater than in group A (20+/-5 versus 15+/-3 mm and 7+/-1 versus 5+/-2 points, respectively). SERCA2 mRNA levels were significantly lower in group B (SERCA2/GAPDH ratio 0.34+/-0.15) compared with group A (0.72+/-0.27) and control subjects (0.85+/-0.47), whereas the mRNA expression of ryanodine receptor-2, phospholamban, calsequestrin, and Na(+)/Ca(2+) exchanger were similar in all groups. CONCLUSIONS: These results suggest that downregulation of SERCA2 mRNA, resulting in altered Ca(2+) handling, may contribute to impaired LV contractile reserve in HCM patients with severe hypertrophy, even in the absence of detectable baseline systolic dysfunction. PMID- 11489772 TI - Catheter ablation in patients with multiple and unstable ventricular tachycardias after myocardial infarction: short ablation lines guided by reentry circuit isthmuses and sinus rhythm mapping. AB - BACKGROUND: Extensive lines of radiofrequency (RF) lesions through infarct (MI) can ablate multiple and unstable ventricular tachycardias (VTs). Methods for guiding ablation that minimize unnecessary RF applications are needed. This study assesses the feasibility of guiding RF line placement by mapping to identify a reentry circuit isthmus. METHODS AND RESULTS: Catheter mapping and ablation were performed in 40 patients (MI location: inferior, 28; anterior, 7; and both, 5) with an electroanatomic mapping system to measure the infarct region and ablation lines. The initial line was placed in the MI region either through a circuit isthmus identified from entrainment mapping or a target identified from pace mapping. A total of 143 VTs (42 stable, 101 unstable) were induced. An isthmus was identified in 25 patients (63%; 5 with only stable VTs, 5 with only unstable VTs, and 15 with both VTs). Inducible VTs were abolished or modified in 100% of patients when the RF line included an isthmus compared with 53% when RF had to be guided by pace mapping (P=0.0002); those with an isthmus identified received shorter ablation lines (4.9+/-2.4 versus 7.4+/-4.3 cm total length, P=0.02). During follow-up, spontaneous VT decreased markedly regardless of whether an isthmus was identified. VT stability and number of morphologies did not influence outcome. CONCLUSIONS: A 4- to 5-cm line of RF lesions abolishes all inducible VTs in more than 50% of patients. Less ablation is required if a reentry circuit isthmus is identified even when multiple and unstable VTs are present. PMID- 11489773 TI - Comparison of right and left ventricular responses to left ventricular assist device support in patients with severe heart failure: a primary role of mechanical unloading underlying reverse remodeling. AB - BACKGROUND: Left ventricular assist devices (LVAD) reverse ventricular, myocardial, and systemic abnormalities characteristic of severe heart failure (reverse remodeling). The relative contributions of hemodynamic unloading and normalized biochemical milieu to reverse remodeling are unknown. METHODS AND RESULTS: Structural and functional characteristics were measured from 53 hearts of patients undergoing transplantation without LVAD support (medical support) and 33 hearts from patients receiving a median of 46 days of LVAD support (range, 8 to 360 days). Compared with medical support alone, patients receiving LVAD support for >/=30 days had higher central venous pressures (11+/-6 versus 8+/-5 mm Hg, P=0.04), lower pulmonary artery diastolic pressures (14+/-9 versus 21+/-9 mm Hg, P=0.01), and higher cardiac outputs (5.1+/-1.6 versus 3.7+/-1.0 L/min, P<0.001). In LVAD versus transplantation hearts, V(30) (ex vivo volume yielding ventricular pressure of 30 mm Hg) was decreased in the left ventricle (LV) (179+/ 75 versus 261+/-118 mL, P=0.005) but not in the right ventricle (RV) (140+/-59 versus 148+/-52 mL, P=NS). LV myocyte diameter decreased more significantly after LVAD support (17%, P=0.05) than in the RV (11%, P=NS). Compared with transplantation, LVAD support increased normalized SERCA2a content in the LV (0.51+/-0.26 versus 1.04+/-0.34, P<0.001) but not in the RV (0.48+/-34 versus 0.67+/-0.55, P=NS). Finally, LVAD support improved force-frequency relations of isolated superfused LV trabeculae (P=0.01) but not RV trabeculae. CONCLUSIONS: Reduction of hemodynamic load is a primary factor underlying several important features of reverse remodeling. These findings do not preclude a possible primary role of neurohormonal factors underlying other facets of reverse remodeling during LVAD support. PMID- 11489774 TI - Cardiac hypertrophy after transplantation is associated with persistent expression of tumor necrosis factor-alpha. AB - BACKGROUND: The mechanisms that contribute to cardiac allograft hypertrophy are not known; however, the rapid progression and severity of hypertrophy suggest that nonhemodynamic factors may play a contributory role. Tumor necrosis factor alpha (TNF-alpha) is a cytokine produced in cardiac allografts and capable of producing hypertrophy and fibrosis; therefore, we suggest that TNF-alpha may play a contributory role. Accordingly, the aims of our study were to define the role of systemic hypertension in the development of hypertrophy, characterize the histological determinants of hypertrophy, and characterize the expression of myocardial TNF-alpha after heart transplantation. METHODS AND RESULTS: To separate the effect of hypertension from immune injury in the development of cardiac allograft hypertrophy, we measured the gain in left ventricular mass by 2D echocardiography in heart transplant recipients and lung transplant recipients who developed similar rates of systemic hypertension. The gain in left ventricular mass was 73% in heart transplant recipients and 7% in lung transplant recipients (P<0.0001). By comparing myocardial samples obtained during the first week after transplant and at 1 year, we found that there was a significant increase in total collagen content (P<0.0001), collagen I (P<0.0001), collagen III (P<0.0001), and myocyte size (P<0.0001). These changes were associated with persistent myocardial TNF-alpha expression. CONCLUSIONS: We suggest that the contribution of hypertension to cardiac allograft hypertrophy is minimal and that persistent intracardiac expression of TNF-alpha may contribute to the development of cardiac allograft hypertrophy. PMID- 11489775 TI - Predictors of outcome of biventricular repair in infants with multiple left heart obstructive lesions. AB - BACKGROUND: Decisions regarding surgical strategy in patients with multiple left heart obstructive or hypoplastic lesions often must be made in the newborn period and are seldom reversible. Predictors of outcome of biventricular repair have not been well defined in this heterogeneous group of patients, and risk factors described for critical aortic valve stenosis have been shown to be inapplicable to patients with other left heart obstructive lesions. The goal of this study was to identify echocardiographic predictors of outcome of biventricular repair for infants with multiple left heart obstructive lesions. METHODS AND RESULTS: Patients with >/=2 areas of left heart obstruction or hypoplasia, diagnosed at 1 L/min. SMVs without clenbuterol administration failed. Thus, we expected that clenbuterol-supported SMVs might become integrated into the circulation by a 1-step operation instead of the 2-step procedure required up to now. METHODS AND RESULTS: In adult Boer goats (n=5), latissimus dorsi muscle was wrapped around a polyurethane chamber of 150 mL that was connected to the descending aorta. This muscular flow-through pumping chamber containing a stabilizing inner layer (called a biomechanical heart [BMH]) was formed and immediately made to work against a systemic load with the support of clenbuterol (5x150 microg/wk). During surgery, the mean stroke volume of BMHs was 53.8+/-22.4 mL. One month after surgery, in peripheral arterial pressure, the mean diastolic (P(MD)) and minimal diastolic (P(min)) pressures of BMH-supported heart cycles differed significantly from unsupported ones (P(MD)=+2.9+/-1.1 mm Hg [P<0.04], P(min)=-2.4+/-0.9 mm Hg [P<0.04]). After BMH-supported heart contractions, the subsequent maximal rate of pressure generation, dP/dt(max), increased by 20.5+/ 8.1% (P<0.02). One BMH, catheterized 132 days after surgery, shifted a volume of 34.8 mL per beat and 1.4 L/min with a latissimus dorsi muscle of 330 g. Depending on duration of training, the percentage of myosin heavy chain type 1 ranged between 31% and 100%. CONCLUSIONS: Under support of clenbuterol, BMHs of a clinically relevant size can be trained effectively in the systemic circulation after a 1-step operation and offer the prospect of a sufficient volume shift and probably unloading of the left ventricle. PMID- 11489782 TI - Electrical induction of ventricular fibrillation for resuscitation from postcountershock pulseless and asystolic cardiac arrests. AB - BACKGROUND: There is increasing evidence that defibrillation from prolonged ventricular fibrillation (VF) before CPR decreases survival. It remains unclear, however, whether harmful effects are due primarily to initial countershock of ischemic myocardium or to resultant postdefibrillation rhythms (ie, pulseless electrical activity [PEA] or asystole). METHODS AND RESULTS: We induced 15 dogs into 12 minutes of VF and randomized them to 3 groups. Group 1 was defibrillated at 12 minutes and then administered advanced cardiac life support (ACLS); group 2 was allowed to remain in VF and was subsequently defibrillated after 4 minutes of ACLS; group 3 was defibrillated at 12 minutes, electrically refibrillated, and then defibrillated after 4 minutes of ACLS. All group 1 and 3 animals were defibrillated into PEA/asystole at 12 minutes. After 4 minutes of ACLS, group 2 and 3 animals were effectively defibrillated into sinus rhythm. The extension of VF in group 2 and 3 subjects paradoxically resulted in shorter mean resuscitation times (251+/-15 and 245+/-7 seconds, respectively, versus 459+/-66 seconds for group 1; P<0.05) and improved 1-hour survival (10 of 10 group 2 and 3 dogs versus 1 of 5 group 1 dogs; Fisher's exact, P<0.005) compared with more conservatively managed group 1 subjects. CONCLUSIONS: Precountershock CPR during VF appears more conducive to resuscitation than CPR during postcountershock PEA or asystole. The intentional induction of VF may prove useful in the management of PEA and asystolic arrests. PMID- 11489783 TI - Upregulation of the cardiac monocarboxylate transporter MCT1 in a rat model of congestive heart failure. AB - BACKGROUND: Cardiac metabolism becomes more dependent on carbohydrates in congestive heart failure (CHF), and lactate may be used as an important respiratory substrate. Monocarboxylate transporter 1 (MCT1) promotes cotransport of lactate and protons into and out of heart cells and conceivably flux of lactate between cells, because it is abundantly present in the intercalated disk. METHODS AND RESULTS: Six weeks after induction of myocardial infarction (MI) in Wistar rats, left ventricular end-diastolic pressures were >15 mm Hg, signifying CHF. MCT1 and connexin43 protein levels in CHF were 260% and 20%, respectively, of those in sham-operated animals (Sham), and the corresponding mRNA signals were 181% and not significantly changed, respectively. Confocal laserscan immunohistochemistry and quantitative immunogold cytochemistry showed that MCT1 density was much higher in CHF than in Sham both at the surface membrane and in the intercalated disk. In CHF, a novel intracellular pool of MCT1 appeared to be associated with cisternae, some close to the T tubules. In contrast, connexin43 particles, seen exclusively at gap junctions, were substantially fewer. Maximum lactate uptake was 107+/-15 mmol. L(-1). min(-1) in CHF and 42+/-6 mmol. L(-1). min(-1) in Sham cells (P<0.05). The K(m) values were between 7 and 9 mmol/L (P=NS). CONCLUSIONS: In cardiomyocytes from CHF rats, (1) the amount of functional MCT1 in the sarcolemma, including in the intercalated disk, is increased several-fold; (2) a new intracellular pool of MCT1 appears; (3) another disk protein, connexin43, is much reduced; and (4) increased reliance on lactate and other monocarboxylates (eg, pyruvate) could provide tight metabolic control of high-energy phosphates. PMID- 11489784 TI - Microcirculation in hypertension: a new target for treatment? PMID- 11489786 TI - Platypnea-orthodeoxia due to aortic elongation. PMID- 11489787 TI - Giant left atrium. PMID- 11489788 TI - Combining cytotoxics and 17-allylamino, 17-demethoxygeldanamycin: sequence and tumor biology matters. Commentary re: P. Munster et al., Modulation of Hsp90 function by ansamycins sensitizes breast cancer cells to chemotherapy-induced apoptosis in an RB- and schedule-dependent manner. Clin. Cancer Res., 7: 2228 2236, 2001. PMID- 11489789 TI - Cancer genomics: promises and complexities. AB - The impending final deciphering of the complete human genome, coupled with the advancement of high-throughput technologies, is positioned to bring about a fundamental transformation in cancer research. The era of molecular biology is transforming into the era of genomic biology, with an unprecedented promise of understanding multifactorial diseases and of identifying specific targets that can be used to develop patient-tailored therapies. Although the genomic approach is in an early phase of its development and its tools need to be honed, the application of genomic technologies to cancer research has already generated exciting results both in target identification and in disease classification. In this article, we review some of the developments pertinent to cancer research, discuss potentially problematic areas associated with them, and comment on future trends and issues. PMID- 11489790 TI - Cell cycle-mediated drug resistance: an emerging concept in cancer therapy. AB - The concept of combining chemotherapeutic agents to increase cytotoxic efficacy has evolved greatly over the past several years. The rationale for combination chemotherapy has centered, in the past, on attacking different biochemical targets, overcoming drug resistance in heterogeneous tumors, and by taking advantage of tumor growth kinetics with increasing the dose-density of combination chemotherapy. The overall goal was to improve clinical efficacy with acceptable clinical toxicity. With our increased understanding of the cell cycle and the impact chemotherapeutic agents have on the cell cycle, it is increasingly apparent that this physiology can create drug resistance, thereby reducing combination chemotherapeutic efficacy. This is particularly relevant with the advent of cell cycle-specific inhibitors but also has relevance for the action of standard chemotherapeutic agents currently in clinical practice. This cell cycle mediated resistance may be overcome by a greater understanding of chemotherapeutic cell cycle effects and by appropriate sequencing and scheduling of agents in combination chemotherapy. In this review, we have elected to illustrate the evolving concept of cell cycle-mediated drug resistance with novel drug combinations that include the taxanes, camptothecins, and fluorouracil. This review indicates that as our understanding of the cell cycle grows, our ability to appropriately sequence chemotherapy to overcome cell cycle-mediated drug resistance can have a great impact on our therapeutic approach in the treatment of human cancers. PMID- 11489791 TI - Clinical pharmacokinetics and metabolism of irinotecan (CPT-11). AB - CPT-11 belongs to the class of topoisomerase I inhibitors, and it acts as a prodrug of SN-38, which is approximately 100-1000-fold more cytotoxic than the parent drug. CPT-11 has shown a broad spectrum of antitumor activity in preclinical models as well as clinically, with responses observed in various disease types including colorectal, lung, cervical, and ovarian cancer. The pharmacokinetics and metabolism of CPT-11 are extremely complex and have been the subject of intensive investigation in recent years. Both CPT-11 and SN-38 are known in an active lactone form and an inactive carboxylate form, between which an equilibrium exists that depends on the pH and the presence of binding proteins. CPT-11 is subject to extensive metabolic conversion by various enzyme systems, including esterases to form SN-38, UGT1A1 mediating glucuronidation of SN-38, as well as CYP3A4, which forms several pharmacologically inactive oxidation products. Elimination routes of CPT-11 also depend on the presence of drug-transporting proteins, notably P-glycoprotein and canalicular multispecific organic anion transporter, present on the bile canalicular membrane. The various processes mediating drug elimination, either through metabolic breakdown or excretion, likely impact substantially on interindividual variability in drug handling. Strategies to individualize CPT-11 administration schedules based on patient differences in enzyme or protein expression or by coadministration of specific agents modulating side effects are under way and may ultimately lead to more selective chemotherapeutic use of this agent. PMID- 11489792 TI - Current clinical and laboratory strategies to augment the efficacy of immunotherapy in multiple myeloma. AB - Multiple myeloma is still an incurable, lethal disease for the vast majority of patients. Myeloablative chemotherapy combined with autologous or allogeneic hematopoietic stem cell transplantation only partially met the great expectations initially set in its efficacy and is associated with a high level of toxicity. However, the considerable progress in understanding the biology of multiple myeloma led to the development of promising molecular therapies. Numerous immunotherapy-based approaches are currently evaluated in clinical trials. Moreover, remarkable progress has been achieved in gene therapy during the last decade, and the repertoire of gene transfer techniques can be expected to improve continuously. Gene transfer is increasingly applied in biological therapies in multiple myeloma. This article reviews the currently applied clinical and laboratory strategies to augment the efficacy of immunotherapy in multiple myeloma and aims to define its perspectives in multimodality treatment of multiple myeloma. PMID- 11489793 TI - Influence of cellular factors and pharmacokinetics on the formation of platinum DNA adducts in leukocytes of children receiving cisplatin therapy. AB - The formation of platinum (Pt)-DNA adducts is thought to be crucial to the antitumor activity of cisplatin, and relationships between adduct formation in peripheral blood leukocytes (PBLs) and response to cisplatin therapy have been reported. The current study directly tests, for the first time, whether pharmacokinetic or other factors predominantly determine the drug-target interaction of cisplatin in a pediatric patient population. Cisplatin pharmacokinetics and Pt-DNA adduct formation in PBLs were determined in 10 children in parallel with measurement of adduct levels after incubation of pretreatment blood samples with cisplatin in vitro. Total and unbound plasma Pt concentrations were determined by atomic absorption spectrophotometry and adduct measurements performed by competitive ELISA. Pt-DNA adduct levels determined after cisplatin treatment showed considerable interindividual variation (peak levels at 24 h ranged from 0.15 to 1.31 nmol/g DNA) and correlated strongly with adduct levels determined after incubation of pretreatment whole blood with cisplatin (r = 0.92; P = 0.0002). No significant correlation was observed between in vivo adduct formation and either unbound or total cisplatin plasma concentrations (r = 0.14 and 0.18, respectively). A correlation was also observed between the degree of myelosuppression, as determined by WBC nadirs measured over a 14-day period after cisplatin treatment, and the extent of adduct formation, with greater WBC toxicity observed in patients with higher levels of Pt-DNA adducts (P = 0.010). These preliminary results provide evidence that interpatient variation in formation of Pt-DNA adducts in PBLs of children is determined by host-specific factors other than cisplatin pharmacokinetics. These results imply that analysis of adducts in PBLs after incubation of pretreatment blood samples with cisplatin may be used to predict in vivo adduct levels, leukopenia, and, potentially, response to cisplatin therapy. PMID- 11489794 TI - Identification of differentially expressed genes in esophageal squamous cell carcinoma (ESCC) by cDNA expression array: overexpression of Fra-1, Neogenin, Id 1, and CDC25B genes in ESCC. AB - PURPOSE: This study aims to identify differentially expressed genes in esophageal squamous cell carcinoma (ESCC) through the use of a membrane-based cDNA array. EXPERIMENTAL DESIGN: Two newly established human ESCC cell lines (HKESC-1 and HKESC-2) and one corresponding to a morphologically normal, esophageal epithelium tissue specimen, prospectively collected from the HKESC-2-related patient, were screened in parallel using a cDNA expression array containing gene-specific fragments for 588 human genes spotted onto nylon membranes. RESULTS: The results of cDNA expression array showed that 53 genes were up-regulated 2-fold or higher and 8 genes were down-regulated 2-fold or higher in both ESCC cell lines at the mRNA level. Semiquantitative RT-PCR analysis of a subset of these differentially expressed genes gave results consistent with cDNA array findings. Four of the differentially expressed genes that belong to the categories of oncogenes/tumor suppressor genes (Fra-1 and Neogenin) and cell cycle-related genes (Id-1 and CDC25B) were studied more extensively for their protein expression by immunohistochemistry. The two ESCC cell lines and their corresponding primary tissues, 61 primary ESCC resected specimens and 16 matching, morphologically normal, esophageal epithelium tissues were analyzed. The immunostaining results showed that Fra-1, Neogenin, Id-1, and CDC25B were overexpressed in both ESCC cell lines and their corresponding primary tumors at the protein level, validating the microarray findings. The results of the clinical specimens showed that the Fra-1 gene was overexpressed in ESCC compared with normal esophageal epithelium in 53 of 61 cases (87%), Neogenin in 57 of 61 cases (93%), Id-1 in 57 of 61 cases (93%), and CDC25B in 48 of 61 cases (79%). Furthermore, the expression of Fra-1, Neogenin, and Id-1 in ESCC correlated with tumor differentiation. CONCLUSIONS: Overall, this study demonstrates that multiple genes are differentially expressed in ESCC and provides the first evidence that oncogenes Fra-1 and Neogenin and cell cycle-related genes Id-1 and CDC25B are overexpressed in ESCC. PMID- 11489795 TI - p53 mutation in plasma DNA and its prognostic value in breast cancer patients. AB - PURPOSE: Tumor-specific DNA has recently been detected in the plasma of lung, head and neck, breast, and colon cancer patients. Detection of tumor-specific genetic materials in cancer patients at sites distant from the tumor, such as in the blood, may provide a unique and valuable tumor marker for diagnosis and prognosis. EXPERIMENTAL DESIGN: The present investigation was aimed at determining the presence of p53 mutations in the peripheral blood of breast cancer patients and its prognostic value in these patients. RESULTS: In this study, we found that the mean concentration of plasma DNA in healthy women was 21 ng/ml, whereas in patients with breast cancer the mean concentration was 211 ng/ml (P < 0.01). p53 mutations were detected in the primary tumors of 46 of 126 (36.5%) breast cancer patients. Of these 46 patients, 30 (65.1%) were found to have p53 mutations in their plasma DNA. p53 mutations in plasma DNA were strongly correlated with clinical stage, tumor size, lymph node (LN) metastasis, and estrogen receptor status (P < 0.05). After a median follow-up of 29 months, univariate and multivariate analysis revealed that both primary tumor and plasma DNA p53 mutations were significant prognostic factors for both relapse-free and overall survival. Furthermore, we demonstrated that patients with both primary tumor and plasma p53 mutations have the worst survival. This outcome occurs in both LN-positive and LN-negative groups. Thirteen of the 22 (59%) patients with recurrence and/or metastasis later had detectable p53 mutations in their plasma DNA. CONCLUSIONS: Detection of p53 mutations in plasma DNA may be used as a prognostic factor and an early marker to indicate recurrence or distant metastasis. PMID- 11489796 TI - Modulation of Hsp90 function by ansamycins sensitizes breast cancer cells to chemotherapy-induced apoptosis in an RB- and schedule-dependent manner. See: E. A. Sausville, Combining cytotoxics and 17-allylamino, 17-demethoxygeldanamycin: sequence and tumor biology matters, Clin. Cancer Res., 7: 2155-2158, 2001. AB - 17-allyl-aminogeldanamycin (17-AAG) is an ansamycin antibiotic that binds to a highly conserved pocket in the Hsp90 chaperone protein and inhibits its function. Hsp90 is required for the refolding of proteins during cellular stress and the conformational maturation of certain signaling proteins. 17-AAG has antitumor activity in cell culture and animal xenograft models and is currently in clinical trial. It causes an RB-dependent G(1) arrest, differentiation, and apoptosis. RB negative cells arrest in mitosis and undergo apoptosis. Hsp90 plays an important role in the cellular response to environmental stress. Therefore, we tested whether the regulation of Hsp90 function by 17-AAG could sensitize cells to cytotoxic agents. 17-AAG sensitized tumor cells to Taxol and doxorubicin. Taxanes cause growth arrest in mitosis and apoptosis. The addition of 17-AAG to cells after exposure to Taxol significantly increased both the activation of caspases 9 and 3 and apoptosis. In cells with intact RB, exposure to 17-AAG before Taxol resulted in G(1) arrest and abrogated apoptosis. Schedule dependence was not seen in cells with mutated RB, because both agents blocked cells in mitosis. Schedule- or RB-dependence was also not observed when cells were treated with 17-AAG and doxorubicin, a DNA-intercalating agent that acts on different phases of the cell cycle. These findings suggest that inhibition of Hsp90 function by 17-AAG enhances the apoptotic effects of cytotoxic agents. The sequence of drug administration and the RB status significantly influence efficacy. PMID- 11489797 TI - Pan-trk inhibition decreases metastasis and enhances host survival in experimental models as a result of its selective induction of apoptosis of prostate cancer cells. AB - During the progression of prostate cancer, molecular changes occur resulting in the autocrine production of a series of neurotrophins by the malignant cells. This is coupled with expression of high-affinity cognate receptors for these ligands, termed trk receptors, by these cancer cells. The binding of the neurotrophins to their trk receptors activates the receptor's latent tyrosine kinase activity inducing a series of signal transduction pathways within these prostate cancer cells. These molecular changes result in the acquisition by prostate cancer cells of a restricted requirement for these trk signaling pathways for optimal survival. CEP-701 is an indolocarbazole compound specifically designed as a potent inhibitor (IC(50), 4 nM) of the tyrosine kinase activity of the trk receptors required for initiation of these survival pathways. In the present studies, the consequences of CEP-701 inhibition of these trk signaling survival pathways were tested in vivo using both rat (R3327 AT 6.3 and H) and human (TSU-pr1 and CWR-22Rv1) prostatic cancer models. These in vivo studies demonstrated that treatment with CEP-701 inhibits the growth of both rodent and human prostate cancers, without being toxic to the normal tissue including the host prostate. Because of this selective effect, CEP-701 inhibits metastasis and growth of both primary and metastatic sites of prostate cancer. Based upon this profile, long-term survival studies were performed using the slow growing Dunning H rat prostate cancer model. For these latter studies, the dosing regimen was 10 mg CEP-701/kg/dose twice a day via gavage 5 days a week. This regimen maintains CEP-701 tumor tissue concentrations of 25-50 nM. Such chronic dosing increased (P < 0.001) the median survival of rats bearing the slow growing H prostate cancers from 408 days (395-432 days, 95% confidence interval) for the vehicle group (n = 18) to 566 days (497-598 days, 95% confidence interval) for the CEP-701-treated group (n = 24). PMID- 11489798 TI - Phase I trial of gemcitabine combined with radiation for the treatment of locally advanced pancreatic adenocarcinoma. AB - Gemcitabine has modest activity in the treatment of advanced pancreatic cancer and is a potent radiosensitizer. We conducted a Phase I trial to determine the maximum tolerated dose of weekly gemcitabine delivered concurrently with radiation therapy for the treatment of locally advanced adenocarcinoma of the pancreatic head and to assess the treatment-related toxic effects associated with such a regimen. Eighteen patients with pathologically proven, locally advanced adenocarcinoma of the pancreatic head were enrolled in this study. Patients received seven weekly doses of gemcitabine with 3000 cGy of external beam radiation therapy delivered during the first 2 weeks of therapy. Six patients received gemcitabine at 350 mg/m(2)/week, nine at 400 mg/m(2)/week, and three at 500 mg/m(2)/week. Grade 3-4 hematological toxicity was observed in over half the patients treated. Nonhematological toxicities were significant and included fatigue, anorexia, nausea, vomiting, and dehydration. Forty-four % of the patients required admission to the hospital for management of nausea/vomiting and dehydration. The risk of hospitalization appeared to be dose-related; all of the three patients treated at 500 mg/m(2)/week required hospital admission during treatment. Seventeen patients were evaluated for response, and eight patients (47%) had evidence of a local anticancer effect. Four of these eight patients (24%) had a partial response to therapy. The median survival for the entire group was 6 months. The 1-year survival rate for patients with an objective response to therapy was 66%. The clinical responses observed in this group of patients suggest gemcitabine is a clinically relevant radiosensitizer in patients with pancreatic adenocarcinoma. However, the toxic effects are significant, suggesting that until dose and scheduling issues are explored further, concomitant administration of gemcitabine and radiation therapy should still be considered investigational. PMID- 11489799 TI - Dose-reduced conditioning and allogeneic hematopoietic stem cell transplantation from unrelated donors in 42 patients. AB - PURPOSE: A fludarabine-based "nonmyeloablative" preparative regimen was investigated in 42 patients with hematological malignancies receiving hematopoietic stem cell grafts from unrelated volunteer donors. EXPERIMENTAL DESIGN: Recipient conditioning consisted of fludarabine 30 mg/m(2) on days -6 to 2 and i.v. busulfan 3.3 mg/kg on days -6 to -5. Antithymocyte globuline was added at 2.5 mg/kg i.v. on days -5 to -2. The patients were grafted with bone marrow (n = 13) or peripheral blood stem cells either unmanipulated (n = 20) or CD34+ selected (n = 9). Graft-versus-host disease prophylaxis was performed with cyclosporine A (CsA, n = 12), CsA/methotrexate (n = 12), or CsA/mycophenolate mofetil (n = 18). RESULTS: With a median follow-up of 13 months (range, 5-26 months), the actuarial disease-free survival is 64% and 38% for patients with lymphoid malignancies and standard-risk leukemia compared with only 14% for patients with high-risk disease. The main cause of treatment failure was relapse of disease in high-risk patients (n = 14). An increased incidence of primary (n = 1) or secondary graft-failure (n = 8) was observed (21%). Chimerism analysis of CD56+/CD3--sorted natural killer (NK) cells, available in 10 patients, showed an impaired increase of donor NK cell chimerism between day 10 and 30 after transplantation in three of four patients with graft failure, whereas the percentage of donor NK cells surpassed 75% in all of the six patients with stable engraftment. CONCLUSIONS: Unrelated transplants after dose-reduced conditioning are associated with a higher risk of graft-failure. Pretransplant host immunosuppression has to be optimized to overcome resistance to grafts from unrelated donors after nonmyeloablative conditioning therapy. PMID- 11489800 TI - Classic Kaposi's sarcoma associated with human herpesvirus 8 infection in a 13 year-old male: a case report. AB - PURPOSE: Classic Kaposi's sarcoma (KS) is rare in children. Although its etiology is not fully understood, human herpesvirus 8 (HHV-8) is present in the angiogenic lesions. We report an HIV-negative, 13-year-old patient of Sicilian descent with HHV-8-associated classic KS to facilitate the diagnosis and treatment of this entity in children. EXPERIMENTAL DESIGN: DNA was extracted from the skin specimen of the patient and analyzed via PCR assay and Southern blot hybridization for HHV 8 DNA. The amplified HHV-8 DNA was cloned, sequenced, and compared with the prototype HHV-8-KS330/BAM. RESULTS: The patient presented with purpuric lesions on the distal lower extremities and the tip of his nose, associated with thrombocytopenia and leukopenia, suggesting an immune-mediated cytopenia. While on prednisone, he developed marked vascular proliferation in the groins. Biopsy of the skin lesions showed KS, and HHV-8 was detected in the tissues by PCR. Sequence analysis of the amplified DNA was homologous to the prototype HHV-8 KS330/BAM. His HHV-8 strain was the A subgroup, the type associated with Mediterranean classic KS. Stopping prednisone and treatment with IFN-alpha and IgG resulted in regression of the groin lesions. CONCLUSIONS: This report emphasizes the importance of recognizing classic KS in children and avoiding immunosuppressive therapies in indolent classic KS. The diagnostic and therapeutic strategies were effective and well tolerated. PMID- 11489801 TI - Meningioma treated with interferon-alpha, evaluated with [(11)C]-L-methionine positron emission tomography. AB - PURPOSE: In meningioma patients with postoperative residual masses, recurrent or primarily inoperable tumors, positron emission tomography (PET) with [(11)C]-L methionine was used to evaluate treatment efficacy of IFN-alpha. EXPERIMENTAL DESIGN: Twelve patients were treated with IFN-alpha at a dose of 1.5-5 million IU s.c. daily. PET, computed tomography, and/or magnetic resonance imaging were performed in all patients before and, at regular intervals, during IFN-alpha treatment. The ratio of tumor hot-spot uptake to cerebellar uptake or to cortex uptake was calculated. This ratio estimates the relative methionine accumulation in the tumor and presumably the proliferative activity in the tumor. RESULTS: During IFN-alpha treatment, PET demonstrated a mean relative percentage of reduction in the uptake ratio (MRelR) of 22.3% in the meningiomas. In nine patients who were considered responders, defined as patients with a positive MRelR, the MRelR was 30.4%. For the three nonresponders, defined as patients with a negative MRelR, the MRelR was -1.8%. Three patients were followed for a long time: two patients for 8 years and one patient for 4 years and 6 months; the two patients followed for 8 years are still on IFN. The volumes of these tumors were constant or showed a slight decrease. No correlation was found between histopathological diagnosis (PAD) WHO grading I-III of meningiomas and response to IFN-alpha treatment. CONCLUSIONS: PET was judged a useful method to predict which patients are suitable for long-term treatment with IFN-alpha and also for dose finding. In five patients treated from 9 months to 8 years, IFN-alpha seemed to be an effective oncostatic drug. The clinical usefulness of IFN-alpha, taking adverse reactions into account, must be evaluated in a larger series of patients. PMID- 11489802 TI - Dendritic cell vaccination with MAGE peptide is a novel therapeutic approach for gastrointestinal carcinomas. AB - The MAGE gene is selectively expressed in cancer tissues such as melanoma or gastrointestinal carcinomas, whereas no expression is observed in normal tissues except testis. There are several reports of successful induction of HLA class I restricted antitumor CTLs using MAGE peptides, and some clinical trials with these immunogenic peptides were reported as effective for some patients with malignant melanoma. However, there are no similar studies in gastrointestinal carcinomas, which are important neoplasms. Autologous dendritic cells (DCs) were generated ex vivo and were pulsed with MAGE-3 peptide, depending on the patient's HLA haplotype (HLA-A2 or A24). Patients were immunized with DC pulsed with MAGE-3 peptide every 3 weeks at four times. Twelve patients with advanced gastrointestinal carcinoma (six stomach, three esophagus, and three colon) were treated, and no toxic side effects were observed. Peptide-specific CTL responses after vaccination were observed in four of eight patients. Improvement in performance status was recognized in four patients. Tumor markers decreased in seven patients. In addition, minor tumor regressions evidenced by imaging studies were seen in three patients. These results suggested that DC vaccination with MAGE-3 peptide is a safe and promising approach in the treatment of gastrointestinal carcinomas. PMID- 11489803 TI - Phase II study of direct intralesional gene transfer of allovectin-7, an HLA B7/beta2-microglobulin DNA-liposome complex, in patients with metastatic melanoma. AB - Cutaneous melanoma is one of the most rapidly increasing cancers in the United States. Because of the lack of effective treatment options and toxicities of most chemotherapeutic and radiation regimes, immunotherapies such as vaccination therapy represent an attractive approach for patients with advanced melanoma. The purpose of this study was to evaluate the response rate, time to progression, and survival of patients with metastatic melanoma treated by direct intratumoral injection with Allovectin-7 (a plasmid DNA encoding the genes HLA-B7 and beta2 microglobulin complexed with a cationic lipid mixture, DMRIE/DOPE. Fifty-two patients with metastatic melanoma were enrolled in this Phase II study. Therapy consisted of six intratumoral injections of 10 microg of Allovectin-7 over a 9 week period. Treatment was well tolerated. Treatment-related adverse events were mild to moderate, the most frequent of which were ecchymosis, pruritus (and/or discomfort at the injection site), and pneumothoraces. Regression of the injected lesion was observed in 18% of patients, including one complete response, three partial responses, and five minor responses. An overall response rate of 4% (two partial responses) was documented, and nine patients (18%) maintained stable disease for at least 11 weeks. Six patients remained alive 25.1 to 39.4 months from their first injection, including two patients with local (injected tumor) responses and one patient with an overall disease partial response. This study demonstrates that intratumoral administration of Allovectin-7 in metastatic melanoma is safe and can produce both responses in injected lesions and in overall disease. Clinical trials optimizing patient selection and combining Allovectin-7 with other modalities of therapy are currently ongoing in an effort to improve response rates. PMID- 11489804 TI - A phase I dose escalation and bioavailability study of oral sodium phenylbutyrate in patients with refractory solid tumor malignancies. AB - PURPOSE: Phenylbutyrate (PB) is an aromatic fatty acid with multiple mechanisms of action including histone deacetylase inhibition. Preclinically, PB demonstrates both cytotoxic and differentiating effects at a concentration of 0.5 mM. We conducted a Phase I trial of p.o. PB patients with refractory solid tumor malignancies to evaluate toxicity, pharmacokinetic parameters, and feasibility of p.o. administration. EXPERIMENTAL DESIGN: Twenty-eight patients with refractory solid tumor malignancies were enrolled on this dose-escalation to maximally tolerated dose trial. Five dose levels of PB were studied: 9 g/day (n = 4), 18 g/day (n = 4), 27 g/day (n = 4), 36 g/day (n = 12), and 45 g/day (n = 4). Pharmacokinetic studies were performed and included an p.o. bioavailability determination. Compliance data were also collected. RESULTS: The recommended Phase II dose is 27 g/day. Overall the drug was well tolerated with the most common toxicities being grade 1-2 dyspepsia and fatigue. Nonoverlapping dose limiting toxicities of nausea/vomiting and hypocalcemia were seen at 36 g/day. The p.o. bioavailability of PB was 78% for all dose levels, and the biologically active concentration of 0.5 mM was achieved at all dose levels. Compliance was excellent with 93.5% of all possible doses taken. No partial remission or complete remission was seen, but 7 patients had stable disease for more than 6 months while on the drug. CONCLUSIONS: PB (p.o.) is well tolerated and achieves the concentration in vivo that has been shown to have biological activity in vitro. PB may have a role as a cytostatic agent and should be additionally explored in combination with cytotoxics and other novel drugs. PMID- 11489805 TI - Pharmacodynamic model of topotecan-induced time course of neutropenia. AB - Pharmacodynamic measures of neutropenia, such as absolute neutrophil count at nadir and neutrophil survival fraction, may not reflect the overall time course of neutropenia. We developed a pharmacokinetic-pharmacodynamic model to describe and quantify the time course of neutropenia after administration of topotecan to children and to compare this with nonhuman primates (NHPs) as a potential preclinical model of neutropenia. Topotecan was administered as a 30-min infusion daily for 5 days, repeated every 21 days. As part of a Phase I Pediatric Oncology Group study, topotecan was administered at 1.4 and 1.7 mg/m(2)/day without filgrastim (POG), and at 1.7, 2, and 2.4 mg/m(2)/day with filgrastim (POG+G). In NHPs, topotecan was administered at 5, 10, and 20 mg/m(2)/day without filgrastim. A pharmacokinetic-pharmacodynamic model was fit to profiles of topotecan lactone plasma concentrations and neutrophil survival fraction from cycle 1 and used to calculate topotecan lactone area under the plasma concentration-versus-time curve from 0 to 120 h (AUC(LAC)) and the area between the baseline and treatment related neutrophil survival fraction (ABC) from 0 to 700 h. The mean +/- SD neutrophil survival fraction at nadir for the POG, POG+G, and NHP groups was 0.12 +/- 0.09, 0.11 +/- 0.17, and 0.09 +/- 0.08, respectively (P > 0.05). The mean +/- SD for the ratio of ABC to AUC(LAC) for the POG and NHP groups was 1.02 +/- 0.38 and 0.16 +/- 0.09, respectively (P < 0.05). The model estimate of ABC and the ratio of ABC to AUC(LAC) in children and NHPs may better reflect sensitivity to chemotherapy-induced neutropenia. PMID- 11489806 TI - Temozolomide: the effect of once- and twice-a-day dosing on tumor tissue levels of the DNA repair protein O(6)-alkylguanine-DNA-alkyltransferase. AB - Temozolomide (TMZ) is a methylating agent of the imidotetrazine class, whose cytotoxic product is O(6)-methylguanine DNA adducts, which initiate a futile recycling of the mismatch repair pathway causing DNA strand breaks and apoptotic cell death in mismatch repair proficient cells. The DNA repair protein O(6) alkylguanine DNA alkyltransferase (AGT) repairs these adducts in a suicide manner and reduces the cytotoxic action of TMZ. An antitumor threshold is reached when sufficient adducts are formed by TMZ to inactivate AGT. In this study, we evaluated the relation between TMZ dosing and AGT depletion in patients with deep visceral tumors and in peripheral blood mononuclear cells (PBMCs) to determine whether the dose of TMZ was sufficient to inactivate AGT and lead to therapeutic efficacy. To do so, we compared single dose therapy with a novel twice daily regimen in a laboratory correlate-driven Phase I dose escalation study. p.o. bolus dose TMZ 200 mg/m(2) daily times five was compared with the same bolus on day 1 followed by nine doses at 12-h intervals of 50, 75, 90, or 100 mg/m(2). Dose-limiting toxicity in the bid regimen (grade IV thrombocytopenia and neutropenia) was seen at 100 mg/m(2), cumulative dose 1100 mg/m(2), and the maximum tolerated dose was 1010 mg/m(2). The degree of tumor tissue AGT activity depletion measured in biopsies before and on day 5 of therapy varied widely, between 0 (in 3 patients) and 99% (in 1), with the majority of patients (10 of 15) having 52-84% tumor AGT depletion. In contrast, AGT activity in PBMCs fell rapidly during TMZ administration to undetectable levels in all dosage groups on day 5 but did not correlate with tumor AGT depletion. TMZ pharmacokinetics were dose proportional; no accumulation occurred >5-day period in the bid regimen. Two partial responses were seen, lasting 3 and 4 months. Five additional patients achieved prolonged stabilization of disease for 4-6 monthly cycles. This is the first study to document that at maximum tolerated doses, TMZ depletes PBMC AGT but only partially and variably depletes visceral tumor AGT in most patients, even during twice daily dosing. Drug combinations or schedules designed to maximally deplete tumor AGT might improve TMZ efficacy. PMID- 11489807 TI - Differential degradation rates of inactivated alkyltransferase in blood mononuclear cells and tumors of patients after treatment with O(6)-benzylguanine. AB - O(6)-Alkylguanine-DNA alkyltransferase (AGT) repairs O(6)-alkylating DNA adducts generated by alkylating therapeutic agents. Therefore, AGT activity may be an important marker of tumor and normal tissue sensitivity to chemotherapeutic agents and a predictor for the success of chemotherapeutic regimens. It is rapidly inactivated by O(6)-benzylguanine (BG) that mimics its substrates, O(6) methylguanine and O(6)-chloroethylguanine DNA adducts. In a Phase I clinical trial, BG was given in increasing doses (from 10 to 120 mg/m(2)) by 1-h infusion. We previously reported depletion of AGT activity, and in this report, we demonstrate the relationship between degradation of BG-inactivated AGT protein and the depletion of AGT activity in peripheral blood mononuclear cells (PBMCs) and tumor samples obtained by computed tomography-guided cutting needle biopsy from patients prior to BG and either 2 or 18 h after BG. In PBMCs, BG inactivated AGT activity by over 95-100% at the end of a 1-h infusion, and depletion was maintained for 18 h. In contrast, AGT protein remained almost unchanged for up to 18 h after BG, suggesting that inactivated AGT proteins remain immunoreactive and are not rapidly degraded in PBMCs. In patient tumor biopsies, AGT activity was depleted approximately 90% 2 h after BG. Tumor AGT protein levels were reduced to approximately 40% of pretreatment values when detected by either Western blot or immunohistochemistry staining. In tumor samples obtained 18 h after BG, >95% inactivation of tumor AGT activity was observed at BG doses of 36-80 mg/m(2), and complete depletion of tumor AGT activity occurred at 120 mg/m(2) BG. However, residual AGT protein (5-10% of baseline) was detectable in all tumor samples. Therefore, the degradation of BG-inactivated AGT protein appeared to be much more rapid in tumors than that in PBMCs, which may impact on AGT regeneration rates as well. Because degradation of BG-inactivated AGT takes place slowly, antibody based measurements of AGT protein correlate poorly with depletion of AGT activity immediately after BG. Thus, biochemical activity measurements remain the appropriate monitor of AGT during therapeutic modulation. These data provide the first and conclusive evidence of differential degradation rates of inactivated AGT in PBMCs and tumors of patients after treatment with BG and suggest that immunoreactive AGT measurements in PBMCs are a poor surrogate for AGT activity in tumor tissue. PMID- 11489808 TI - Cisplatin, etoposide, and paclitaxel with granulocyte colony-stimulating factor in untreated patients with extensive-stage small cell lung cancer: a phase II trial of the Southwest Oncology Group. AB - PURPOSE: This study was designed to determine the efficacy and toxicity of cisplatin, etoposide, and paclitaxel (PET) in patients with extensive-stage small cell lung cancer (ES-SCLC). EXPERIMENTAL DESIGN: Chemo-naive adult patients with a performance status (PS) of 0-2 and adequate organ function were eligible. Patients received cisplatin 80 mg/m(2) i.v., etoposide 80 mg/m-2 i.v., and paclitaxel 175 mg/m(2) i.v. over a 3-h period on day 1 followed by etoposide 160 mg/m(2) p.o. on days 2 and 3 every 21 days for six cycles. G-CSF 5 microg/kg was injected s.c. on days 4-14. RESULTS: Eighty-eight patients were assessable. The median age was 60 years; 50% were male, 78% had PS of 0-1, 28% had PS of 2, 53% had multiple sites, and 13% had brain involvement. The overall response rate was 57% with 10 (12%) of 84 patients achieving a complete response. Median progression-free survival was 6 months [95% confidence interval (CI), 5-7 months] with a median survival of 11 months (95% CI, 8-13 months) and a 1-year survival rate of 43% (95% CI, 33-54%). Six patients (7%) died from toxicity. Grade 5 toxicity occurred in 3 (14%) of 22 patients (with a PS of 2) versus 3 (5%) of 61 patients (with a PS of 0-1; P, not significant). Grade 4 neutropenia developed in 40% of patients. Grade 3 nonhematological toxicities were primarily nausea (20%), vomiting (16%), and fatigue (14%). CONCLUSION: The survival result achieved was superior to prior SWOG experiences; however, the toxic death rate was unacceptably high in PS-2 patients. These results provide the largest database for the ongoing randomized Intergroup trial comparing PET to cisplatin+etoposide in PS-0-1 patients with ES-SCLC. PMID- 11489809 TI - Impact of the putative differentiating agent sodium phenylbutyrate on myelodysplastic syndromes and acute myeloid leukemia. AB - Sodium phenylbutyrate (PB) is an aromatic fatty acid with cytostatic and differentiating activity against malignant myeloid cells (ID(50), 1-2 mM). Higher doses induce apoptosis. Patients with myelodysplasia (n = 11) and acute myeloid leukemia (n = 16) were treated with PB as a 7-day continuous infusion repeated every 28 days in a Phase I dose escalation study. The maximum tolerated dose was 375 mg/kg/day; higher doses led to dose-limiting reversible neurocortical toxicity. At the maximum tolerated dose, PB was extremely well tolerated, with no significant toxicities; median steady-state plasma concentration at this dose was 0.29 +/- 0.16 mM. Although no patients achieved complete or partial remission, four patients achieved hematological improvement (neutrophils in three, platelet transfusion-independence in one). Other patients developed transient increases in neutrophils or platelets and decrements in circulating blasts. Monitoring of the percentage of clonal cells using centromere fluorescence in situ hybridization over the course of PB administration showed that hematopoiesis remained clonal. Hematological response was often associated with increases in both colony-forming units-granulocyte-macrophage and leukemic colony-forming units. PB administration was also associated with increases in fetal erythrocytes. These data document the safety of continuous infusion PB and provide preliminary evidence of clinical activity in patients with myeloid malignancies. PMID- 11489810 TI - Increased incidence of luteinizing hormone-releasing hormone receptor gene messenger RNA expression in hormone-refractory human prostate cancers. AB - There are few options for treating hormone-refractory prostate cancer (PC). Various studies indicate that luteinizing hormone-releasing hormone (LHRH) agonists may have a direct inhibitory effect on prostate tumors mediated by specific LHRH receptors. One study evaluated LHRH receptors in hormone-dependent PC tissue, but no data have thus far been obtained on the presence of LHRH receptors in benign prostatic hyperplasia (BPH) and especially hormone-refractory PC in patients. Thus, it is not yet clear whether LHRH receptors indicate tumor related differentiation or even hormone-refractory dedifferentiation or are likewise associated with BPH. The aim of this study was to determine the rate of LHRH receptor mRNA expression in BPH and in primary, potentially androgen dependent and in hormone-refractory PC with clinical progression. Multiplex reverse transcription-PCR was used to simultaneously detect the expression of mRNA for LHRH receptors and beta-actin in 48 patients with BPH, 14 with a primary, possibly hormone-dependent, prostate carcinoma (PPC), and 18 with a hormone-refractory prostate carcinoma (HRPC). Sixteen of 18 samples with HRPC showed intact RNA and expressed mRNA for LHRH receptors (100%). However, the RNA intact PPC and BPH showed significantly lower expression of mRNA for LHRH receptors (46.2 and 55.3%, respectively; variance analysis: P = 0.0017). The significantly higher expression of mRNA for LHRH receptors in HRPC indicates that therapeutic concepts should be developed that target this site of action. In addition to possible direct effects of LHRH agonists or antagonists demonstrated previously in vitro, it seems useful to apply targeted cytotoxic LHRH analogues or monoclonal antibodies. PMID- 11489811 TI - Matrix metalloproteinase-1 promoter polymorphism 1G/2G is correlated with colorectal cancer invasiveness. AB - PURPOSE: Matrix metalloproteinase-1 (MMP-1) is likely to be involved in invasion and metastasis of several tumors by degrading the extracellular matrix. A single guanine insertion polymorphism (2G) in the MMP-1 promoter region creates an Ets binding site causing the elevation of transcriptional level and local expression of MMP-1. The aim of this study was to evaluate the impact of this 2G insertion type polymorphism on invasion and metastasis of colorectal cancer (CRC). EXPERIMENTAL DESIGN: We genotyped for this 1G/2G polymorphism 60 patients, who were operated on for CRC and followed for 6-30 months (median: 21). A control population of 164 age- and sex-matched tumor-free subjects was also genotyped for the same polymorphism. RESULTS: The proportion of 2G homozygotes was higher in the CRC group than in the controls (P = 0.014; odds ratio, 2.21; 95% confidence interval, 1.17-4.16). The CRC group was divided in a group without metastasis (M ) and a group that had developed metastasis (M+). At the time of diagnosis, 2G homozygotes were more represented in the M+ group than in M- (P = 0.0082; odds ratio, 4.73; 95% confidence interval, 1.46-15.26). The difference between M- patients and controls did not achieve statistical significance (P = 0.52). CONCLUSIONS: Our results suggest that the presence of 2G polymorphism at the MMP 1 promoter region may favor the growth and the metastatic process in CRC patients and could be looked at as a risk factor for a worse prognosis. PMID- 11489812 TI - Proliferation, apoptosis, and survival in high-level microsatellite instability sporadic colorectal cancer. AB - Sporadic colorectal cancer (CRC) characterized by high-level DNA microsatellite instability (MSI-H) has a favorable prognosis. The reason for this MSI-H survival advantage is not known. The aim of this study was to correlate proliferation, apoptosis, and prognosis in CRC stratified by MSI status. The proliferative index (PI) was measured by immunohistochemical staining with the Ki-67 antibody in a selected series of 100 sporadic colorectal cancers classified according to the level of MSI as 31 MSI-H, 29 MSI-Low (MSI-L), and 40 microsatellite stable (MSS). The Ki-67 index was significantly higher in MSI-H cancers (P < 0.0001) in which the PI was 90.1 +/- 1.2% (mean +/- SE) compared with 69.5 +/- 3.1% and 69.5 +/- 2.3% in MSI-L and MSS subgroups, respectively. There was a positive linear correlation between the apoptotic index (AI) and PI (r = 0.51; P < 0.001), with MSI-H cancers demonstrating an increased AI:PI ratio indicative of a lower index of cell production. A high PI showed a trend toward predicting improved survival within MSI-H cancers (P = 0.09) but did not predict survival in MSI-L or MSS cancers. The AI was not associated with survival in any MSI subgroup. In conclusion, this is the first study to show that sporadic MSI-H cancers are characterized by a higher AI:PI ratio and increased proliferative activity compared with MSI-L and MSS cancers, and that an elevated PI may confer a survival advantage within the MSI-H subset. PMID- 11489813 TI - A re-evaluation of carcinoembryonic antigen (CEA) as a serum marker for breast cancer: a prospective longitudinal study. AB - PURPOSE: Carcinoembryonic antigen (CEA) is still a widely used test for monitoring breast cancer, although recent reports discourage its routine use because of low sensitivity. This is a prospective study evaluating the efficacy of CEA and CA 15.3 in monitoring breast cancer. EXPERIMENTAL DESIGN: Serum CEA and CA 15.3 were measured in 2191 patients with either benign (n = 738) or malignant (n = 1453) breast diseases. Five hundred and forty-nine patients were monitored during postsurgical follow-up for either a minimum of 5 years or until time of recurrence. Fifty-three patients with metastases were also monitored during chemotherapy. RESULTS: Elevated CEA and CA 15.3 levels were found in 16.7% and 33.0% of patients, respectively. CEA sensitivity rose to 41.3% and CA 15.3 sensitivity rose to 80.8% in metastatic patients. The adjunct of CEA increased the CA 15.3 sensitivity by 6% in the overall population and by only 2.1% for patients with metastases. During postsurgical follow-up, CEA was elevated in 38.0% and CA 15.3 in 70.2% of patients with recurrence. The combination of CEA and CA 15.3 increased the overall sensitivity by only 1.4%. Longitudinal monitoring of 53 metastatic patients undergoing chemotherapy demonstrated that, when positive, both CEA and CA 15.3 paralleled response to treatment, although CA 15.3 was a significantly more powerful marker for determining response to treatment. The cost effectiveness ratio of CEA was clearly less favorable than that of CA 15.3. CONCLUSIONS: CEA monitoring should be considered an expensive and inefficient method of follow-up evaluation for breast cancer patients, and it provides no additional value when used in combination with CA 15.3. PMID- 11489814 TI - Human kallikrein 4 (KLK4) is highly expressed in serous ovarian carcinomas. AB - Previous studies indicated that a new member of the human kallikrein (KLK) gene family, KLK4, was expressed in prostate, breast, and endometrial carcinoma cell lines and may have potential as a tumor marker. The aim of this study was to examine the expression of KLK4 in the normal ovary and ovarian tumors of different histology, stage, and differentiation and to determine its association with ovarian tumor progression. Using reverse transcription-PCR, Southern blot, and densitometry analyses, we found the level of KLK4 expression was higher in late stage serous (SER) epithelial-derived ovarian carcinomas than in normal ovaries, mucinous epithelial tumors, and granulosa cell tumors. KLK4 was highly expressed in all of the SER ovarian carcinoma cell lines (eight of eight), SER epithelial carcinomas (11 of 11), and two adenomas, whereas it was expressed at a lower level (or not at all) in normal ovaries (four of six), mucinous epithelial tumors (three of four), endometrioid carcinomas (four of five), clear cell carcinomas (two of three), or granulosa cell tumors (three of six). Of particular interest, KLK4 mRNA variants were detected in SER ovarian carcinoma cell lines and primary cultured ovarian tumor cells, but they were not present in normal ovaries. In situ hybridization analysis showed that KLK4 mRNA transcripts are localized to adenocarcinoma cells of ovarian tumor tissues. Similarly, immunohistochemical staining of ovarian carcinoma sections showed immunoreactivity to KLK4 protein product (hK4) antipeptide antibodies. In addition, intracellular hK4 levels, as detected on Western blot analysis, were induced by 100 nM estrogen treatment of the estrogen receptor positive ovarian carcinoma cell line OVCAR-3, >8-24 h. Our results show that the level of KLK4 expression and expression of KLK4 mRNA variants are associated with progression of ovarian cancer, particularly late stage SER adenocarcinomas. Moreover, hK4 may be a candidate marker for the diagnosis and/or monitoring of ovarian epithelial carcinomas. PMID- 11489815 TI - Prognostic value of human kallikrein 10 expression in epithelial ovarian carcinoma. AB - PURPOSE: Human kallikrein 10 (hK10; also known as the normal epithelial cell specific 1 gene and protein) is a secreted serine protease, which belongs to the human kallikrein family. It has been reported that hK10 is down-regulated in breast and prostate cancer cell lines and that it may function as a tumor suppressor. Recently, we developed a highly sensitive and specific immunoassay for hK10 and found that this protein is abundantly expressed in ovarian tissue. In this study, we measured quantitatively hK10 levels in ovarian cancer cytosolic extracts and evaluated the prognostic value of this biomarker in ovarian cancer. EXPERIMENTAL DESIGN: Specimens from eight normal ovarian tissues, eight ovarian tissues with benign disease, and 182 ovarian tumors were investigated. RESULTS: hK10 concentration in ovarian tumor cytosols ranged from 0 to 84 ng/mg of total protein, with a median of 2.6. This median was highly elevated in comparison with normal and benign ovarian tissues (P < 0.001). A cutoff of 1.35 ng/mg was selected to categorize tumors as hK10 high and hK10 low. With chi(2) test and Fisher's exact test, high concentration hK10 was found to be associated with advanced disease stage, serous histological type, suboptimal debulking, and large residual tumor (>1 cm; all P < 0.05). hK10 status was additionally correlated with clinical outcome, including progression-free (PFS) and overall survival (OS) using the Cox model. In univariate analysis, we found that patients with hK10 high tumors were more likely to die and relapse, in comparison with patients with hK10 low tumors (hazards ratios for PFS and OS were 1.93 and 2.42, respectively; P < 0.05). Although this correlation disappeared after the entire patient population was subjected to multivariate analysis, it remained significant in the subgroup of patients with stage III/IV ovarian cancer (hazards ratios for PFS and OS were 1.98 and 2.12, respectively; P < 0.05). CONCLUSIONS: Our results indicate that hK10 is a new, independent, unfavorable prognostic marker, especially for late-stage ovarian cancer. PMID- 11489816 TI - Higher human kallikrein gene 4 (KLK4) expression indicates poor prognosis of ovarian cancer patients. AB - PURPOSE: Kallikrein gene 4 (KLK4, also known as prostase/KLK-L1), located on chromosome 19q13.4, is one of the newly discovered members of the human KLK-like gene family. This gene is up-regulated by androgens in the LNCaP prostatic carcinoma cell line and by androgens and progestins in the BT-474 breast cancer cell line. On the basis of its apparent association with hormonally regulated tissues, we have undertaken to examine the prognostic value of KLK4 expression in 147 malignant ovarian tissues. EXPERIMENTAL DESIGN: Tumors were pulverized, total RNA was extracted, and cDNA was prepared by reverse transcription. KLK4 was amplified by PCR using gene-specific primers, and its identity was verified by sequencing. Ovarian tissues were then classified as KLK4-positive or -negative, based on ethidium bromide visualization of the PCR product on agarose gels. RESULTS: KLK4 was found to be expressed in 69 (55%) of 147 of ovarian cancer samples. We found a strong positive association between KLK4 expression and tumor grade (P = 0.02) and clinical stage (P < 0.001). Univariate survival analysis revealed that patients with ovarian tumors positive for KLK4 expression had an increased risk for relapse and death (P = 0.003 and 0.001, respectively). Whereas knowledge of KLK4 status did not significantly increase the prognostic power of the multivariate models, additional analyses did determine that KLK4 was an independent unfavorable prognostic factor in patients with grade 1 and 2 tumors. CONCLUSIONS: Our findings indicate that KLK4 expression is associated with more aggressive forms of ovarian cancer. PMID- 11489817 TI - Prognostic and pathologic significance of quantitative protein expression profiling in human gliomas. AB - PURPOSE: Analysis of tumor-derived genetic lesions has provided insights into molecular pathogenesis of human gliomas. Because these changes represent only one of several mechanisms that alter gene expression during tumorigenesis, it is likely that further information will be obtained from a careful analysis of important regulatory proteins present in these tumors. EXPERIMENTAL DESIGN: We have quantified the levels of key cell cycle/signaling proteins in 94 prospectively collected, meticulously preserved, "snap frozen" glioma specimens and have compared these levels with histopathological data and patient outcome. RESULTS: The results of these experiments confirm that the levels of wild-type tumor suppressor proteins, such as p53, pRB, PTEN, p14(ARF), and p16(INK4), are lost or severely reduced in most gliomas, and that epidermal growth factor receptor, 2human telomerase reverse transcriptase, and cyclin-dependent kinase 4 are overexpressed frequently and with a few exceptions, almost exclusively, in glioblastomas. In addition, we report frequent underexpression of E2F-1 (in 55% of gliomas) and cyclin E overexpression (in 26% of gliomas), which have not yet been reported on the genomic level. Several of these markers significantly correlated with histopathological grade, and the levels of five proteins showed significant association with patient outcome. In particular, overexpression of epidermal growth factor receptor, human telomerase reverse transcriptase, cyclin dependent kinase 4, and cyclin E was largely restricted to glioblastomas and was significantly associated with reduced patient survivals. CONCLUSIONS: We conclude that the quantitation of cell cycle/signaling proteins from meticulously preserved glioma specimens provides further insights into the molecular pathogenesis of human gliomas and yields valuable prognostic information. PMID- 11489818 TI - Increased expression of matrix metalloproteinases (MMP)-2, MMP-9, and the urokinase-type plasminogen activator is associated with progression from benign to advanced ovarian cancer. AB - Proteases are linked to the malignant phenotype of different solid tumors. Therefore, the expression of the matrix metalloproteinase (MMP)-2 and MMP-9 and of the serine protease urokinase-type plasminogen activator (uPA) and its inhibitor plasminogen activator inhibitor type 1 (PAI-1) in the progression of ovarian cancer was investigated. Gelatinolytic activity and protein expression of MMP-2 and MMP-9 were analyzed in tissue extracts of 19 cystadenomas and 18 low malignant potential (LMP) tumors, as well as 41 primary tumors of advanced ovarian cancer stage International Federation of Gynecology and Obstetrics IIIc/IV and their corresponding omentum metastases by quantitative gelatin zymography and Western blot. In the same tissue extracts, antigen levels of uPA and its inhibitor PAI-1 were determined by ELISA. Protein expression of pro-MMP-2 (72 kDa) and pro-MMP-9 (92 kDa as well as antigen levels of uPA and PAI-1 were low in benign ovarian tumors but increased significantly from LMP tumors to advanced ovarian cancers. The highest values of all of the proteolytic factors were detected in omentum metastases. Active MMP-2 enzyme (62 kDa) was detected only in ovarian cancer (66%) and corresponding metastases (93%) but never in benign or LMP tumors. The activation rate of MMP-2 to its active isoform was higher in the metastases. Comparing both proteolytic systems, higher PAI-1 concentrations were consistently found in cancers with high pro-MMP-9 expression. These data indicate that members of the plasminogen activator system, as well as the metalloproteinases MMP-2/9, increase with growing malignant potential of ovarian tumors. These findings are of particular relevance to the development of protease inhibitors as new therapeutic approaches in ovarian cancer. PMID- 11489819 TI - Expression of the Endostatin gene in epithelial ovarian cancer. AB - Endostatin, a M(r) 20,000 COOH-terminal fragment of collagen XVIII, is currently in preclinical development as a novel antiangiogenic agent. The gene expression of this molecule in 23 normal ovaries with follicle or corpus luteum and in 64 cases of epithelial ovarian cancer (27 serous, 18 mucinous, 13 endometrioid, 4 clear cell, and 2 undifferentiated carcinomas) was analyzed by PCR of RNA after reverse transcription. Seven of the cases were of low malignant potential. With regard to staging, 23 cases had stage I disease, 5 had stage II disease, 29 had stage III disease, and 7 had stage IV disease. The level of endostatin gene expression was described in terms of the ratio of the relative yield of the endostatin gene to that of the beta2-microglobulin gene. Endostatin gene expression in ovarian cancers (median, 0.14; range, 0.02-1.11) was significantly higher than that in normal ovaries with follicle or corpus luteum (median, 0.08; range, 0.03-0.26; P = 0.009). International Federation of Gynecology and Obstetrics stage (P = 0.009) and residual tumor (P = 0.005) were significantly associated with endostatin gene expression; however, other clinicopathological features (e.g., patient age at diagnosis, histological subtype, and histological grade) were not significantly associated with endostatin gene expression. Survival data were available for all patients. Univariate Cox regression analysis showed the prognosis of the patients with high endostatin gene expression [equal to or greater than the median (> or =0.14)] to be significantly worse than that of patients with low endostatin gene expression [less than the median (<0.14); P = 0.044]. Our results with regard to the gene expression of this endogenous inhibitor of angiogenesis present a new insight to understand the biology of epithelial ovarian cancer and may lead to the development of a new therapeutic strategy for epithelial ovarian cancer. PMID- 11489820 TI - Genetic changes in paired atypical and usual ductal hyperplasia of the breast by comparative genomic hybridization. AB - PURPOSE: Breast cancer is thought to develop from noninvasive precursor lesions, although the earliest steps of neoplastic transformation are still undefined. Usual ductal hyperplasia (UDH) is considered to represent a benign proliferation of ductal epithelial cells, whereas atypical ductal hyperplasia (ADH) may represent the first clonal neoplastic expansion of these cells. The aim of this study was to examine genetic alterations in UDH and ADH and to determine the relationship between these lesions in the same breast biopsy. EXPERIMENTAL DESIGN: Comparative genomic hybridization analysis was used to define copy number alterations in DNA extracted from archival sections of 18 patients. Nine patients showed ADH with adjacent UDH, and nine showed pure UDH. None showed evidence of invasive cancer or ductal carcinoma in situ. RESULTS: Five of the nine ADH lesions showed chromosome copy number alterations. 16q loss (five cases) and 17p loss (two cases) were the most frequent changes. The associated UDH lesions in these five patients also showed copy number alterations, always a subset of the changes present in the paired ADH. In one other patient, the UDH showed eight chromosomal alterations, whereas the paired ADH showed no changes. Only one of nine cases with pure UDH showed comparative genomic hybridization abnormalities. CONCLUSIONS: These data support the likelihood that UDH is a precursor of ADH, at least in some cases representing neoplastic growth. The frequencies of 16q and 17p losses suggest that alterations of candidate genes located in these chromosomal regions may play a role early in breast carcinogenesis. PMID- 11489821 TI - Gelsolin as a negative prognostic factor and effector of motility in erbB-2 positive epidermal growth factor receptor-positive breast cancers. AB - PURPOSE: erbB-2 and epidermal growth factor receptor (EGFR) may mediate motility via signaling that enables changes in the actin cytoskeleton. A physical basis for this motility may depend on the coexpression of gelsolin, a M(r) 80,000 actin binding protein. EXPERIMENTAL DESIGN: The expression of erbB-2, EGFR, and gelsolin was analyzed in 790 archival invasive breast cancers. These data were compared with histological, clinical, and outcome data (median follow-up, 16.3 years). RESULTS: Protein overexpression was observed in overlapping subsets of breast cancers (38% of cases were erbB-2+; 15% of cases were EGFR+; and 56% of cases were gelsolin+). Tumor gelsolin was associated with overexpression of erbB 2 and EGFR, as well as with an aggressive tumor phenotype. By univariate and multivariate analyses, tumor gelsolin alone was not a prognostic factor. Overexpression of all three factors significantly predicted poor clinical outcome by univariate and multivariate analyses. For example, in node-positive patients, coexpression of all three markers was associated with a 3-year disease-specific survival (as compared with erbB-2+, EGFR+, gelsolin- patients, who had a median survival of 6 years). CONCLUSIONS: These data suggest that gelsolin coexpression may be an important additional prognostic factor in erbB-2+, EGFR+ breast cancer patients. We hypothesize that this is due to the role of gelsolin in mediating motility and invasion. PMID- 11489822 TI - MYC messenger RNA expression predicts survival outcome in childhood primitive neuroectodermal tumor/medulloblastoma. AB - PURPOSE AND EXPERIMENTAL DESIGN: Cerebellar primitive neuroectodermal tumors/medulloblastomas (PNET/MB) are the most common malignant brain tumors in childhood. To identify PNET/MB biological prognostic factors that define a patient group with a sufficiently good prognosis to permit a reduction in treatment intensity, we determined the expression levels of MYC mRNA in fresh frozen tumor samples from 26 PNET/MB patients using semiquantitative reverse transcription-PCR. RESULTS: MYC mRNA expression levels in primary PNET/MB showed a wide range with a 22-fold difference between the highest and lowest values and did not correlate with MYC gene amplification. MYC mRNA expression was an independent significant prognostic factor for progression-free survival outcome and was more predictive than standard clinical factors. The combination of low MYC mRNA expression and high TrkC mRNA expression identified a good outcome group of PNET/MB patients (n = 7) with 100% progression-free survival after a median follow-up time of 55 months (range, 15-91 months). Three of these seven good outcome patients survived without radiotherapy. CONCLUSIONS: Low MYC mRNA expression is a powerful independent predictor of favorable clinical outcome in PNET/MB. Assessment of MYC mRNA levels is feasible and may be incorporated in prospective PNET/MB clinical trials to aid in treatment planning for patients with PNET/MB on confirmation of our results in larger studies. PMID- 11489823 TI - Biological behavior of human breast cancer micrometastases. AB - PURPOSE: Clinically undetectable micrometastases may account for disease recurrence in breast cancer patients after variable disease-free intervals. However, little is known about the cellular mechanisms controlling human breast cancer micrometastases. We compared tumor proliferation rate, apoptotic index, and angiogenesis in human breast cancer micrometastases with those of macroscopic axillary lymph node metastases. EXPERIMENTAL DESIGN: Seven breast cancer micrometastases (<2 mm) obtained from the sentinel nodes of seven patients were compared with 13 macrometastases (lymph node replaced with tumor) obtained from 13 patients. The tissue was fixed in formalin, embedded in paraffin, serially sectioned, and evaluated by H&E and immunohistochemistry for cytokeratin. Tumor proliferation rate was assessed as the number of Ki-67-positive nuclei/total number of tumor nuclei. Tumor vascularity was quantified using antibody to factor VIII to identify microvessels per high-power field (at x400). Apoptosis was quantified using the terminal deoxynucleotidyl transferase (Tdt)-mediated nick end labeling method. Results were analyzed with the Wilcoxon rank-sum test. RESULTS: Median size of micrometastases was 0.5 mm (range, 0.4-1.0), and the median number of tumor nuclei/section was 143 (range, 90-312). Median proliferation rate for macrometastases was greater than for micrometastases (35% versus 12%; P = 0.003). Median microvessel density/high-power field for macrometastases was greater than for micrometastases (17 versus 1; P < 0.001). There was no difference in apoptotic index between macrometastases and micrometastases (1.1% versus 0.7%; P = not significant). CONCLUSIONS: Human breast cancer micrometastases have lower tumor proliferation rates and angiogenesis than breast cancer macrometastases. These characteristics may explain their differential growth patterns. PMID- 11489824 TI - Her-2/neu overexpression in muscle-invasive urothelial carcinoma of the bladder: prognostic significance and comparative analysis in primary and metastatic tumors. AB - PURPOSE: The prognostic significance of Her-2/neu overexpression in muscle invasive urothelial carcinoma of the bladder is largely unknown. Accurate determination of Her-2/neu overexpression may have therapeutic importance. EXPERIMENTAL DESIGN: Eighty consecutive cases of muscle-invasive urothelial carcinoma of the bladder treated by radical cystectomy with available follow-up were analyzed. In each case, one representative section was stained with anti-Her 2/neu. Staining was graded as 1 = faint/equivocal, 2 = moderate, and 3 = strong and was considered positive if > or =2. In those cases with a metastasis, the stain was also performed in the metastatic tumor. Results were correlated with survival. RESULTS: Twenty-two (28%) cases were considered Her-2/neu-positive in the primary tumor, and 17 of 32 (53%) were considered Her-2/neu-positive in the lymph node metastasis. Median survival for Her-2/neu-positive primary tumors was 33 months, compared with 50 months for Her-2/neu-negative cases (P = 0.46). Similarly, Her-2/neu overexpression in the lymph node metastasis did not predict survival. Sixty metastatic urothelial carcinomas were further studied by comparing Her-2/neu expression in the primary tumor with that of the lymph node and/or distant metastasis. Forty-five percent of Her-2/neu-negative primary tumors had a Her-2/neu-positive lymph node metastasis, whereas only one case (8%) of Her-2/neu-positive primary tumors was Her-2/neu-negative in the lymph node metastasis (P = 0.009). Similarly, 67% of Her-2/neu-negative primary tumors had a Her-2/neu-positive distant metastasis, whereas no Her-2/neu-positive primary tumor was negative in the metastasis (P = 0.429). CONCLUSIONS: Her-2/neu overexpression in primary or metastatic tumor did not predict survival in this cohort of muscle-invasive tumors. Overexpression in the primary tumors consistently predicts overexpression in a distant or regional metastasis. However, some Her-2/neu-negative primary tumors may show overexpression in their corresponding metastasis. Her-2/neu analysis in a metastasis may be necessary to accurately determine Her-2/neu status in metastatic bladder urothelial carcinoma. PMID- 11489825 TI - Her-2/neu and urokinase-type plasminogen activator and its inhibitor in breast cancer. AB - PURPOSE: Recent studies suggest that HER-2/neu specifically promotes the invasive capacity of tumor cells by up-regulating secretion of the proteolytic enzyme, urokinase-type plasminogen activator (uPA), or its inhibitor, plasminogen activator inhibitor-1 (PAI-1), in colon and gastric cancer. It was the purpose of this study to: (a) evaluate the association between HER-2/neu and uPA and PAI-1 expression in a large primary breast cancer cohort; (b) perform the first multivariate analysis, including HER-2/neu, uPA, and PAI-1 in breast cancer; and (c) define the effect of HER-2/neu overexpression on uPA and PAI-1 expression in breast cancer cells. EXPERIMENTAL DESIGN: HER-2/neu, uPA, and PAI-1 were measured as continuous variables by ELISA in primary breast cancer tissue extracts from 587 patients with clinical follow-up and analyzed for correlations with clinical outcome. Furthermore, a full-length human HER-2/neu cDNA was introduced into five human breast cancer cell lines to define the effects of HER-2/neu overexpression on uPA and PAI-1 expression. In addition, we tested whether HER-2/neu antibodies could reverse any given alteration of uPA and PAI-1 levels. RESULTS: Our findings indicate a weak positive association between HER-2/neu and uPA (r = 0.147; P < 0.001) and no association between HER-2/neu and PAI-1 (r = 0.07; P = 0.085). HER 2/neu overexpression (> or =400 fmol/mg) and high levels of uPA/PAI-1 (> or =5.5 ng/mg and/or > or =14 ng/mg, respectively) were significantly associated with shorter disease-free survival (DFS; P < 0.001 and P = 0.003) and metastasis-free survival (MFS; P = 0.015 and P < 0.001). Multivariate analysis revealed prognostic independence between HER-2/neu and the uPA/PAI-1 axis for DFS and MFS. Both uPA and PAI-1 had no significant discriminatory effect among HER-2/neu positive patients for DFS. The prognostic value of HER-2/neu overexpression for MFS, however, was significantly enhanced by elevated uPA expression (P = 0.053). Stable transfection of the HER-2/neu gene into multiple human breast cancer cell lines resulted in consistent down-regulation of uPA or PAI-1 expression. In addition, anti-HER-2/neu antibodies did not significantly affect uPA or PAI-1 expression in human cancer cell lines naturally overexpressing HER-2/neu. CONCLUSIONS: The present findings suggest that the invasive phenotype elicited by HER-2/neu overexpression in breast cancer is not a direct effect of uPA or PAI-1 expression. HER-2/neu and the uPA/PAI-1 axis have been shown to affect the invasive capacity of breast cancer independently. Determination of uPA can provide significant additional prognostic information for MFS in HER-2/neu positive and -negative patients. PMID- 11489826 TI - Clinical significance of cyclin B1 protein expression in squamous cell carcinoma of the tongue. AB - PURPOSE: Cyclin B1 plays an important role in control of the G(2)-M transition of the cell cycle. We have shown recently that overexpression of cyclin B1 is associated with poor outcome in patients with early stage squamous cell carcinoma (SCC) of the lung. EXPERIMENTAL DESIGN: To determine the role of cyclin B1 in SCC of the tongue, we analyzed tumor specimens from 41 patients with stage II-IV SCC of the tongue who underwent curative surgery using immunohistochemistry. RESULTS: The median follow-up of all patients was 83 months. Overexpression of cyclin B1 was observed in 15 (37%) of the 41 tumors, a similar frequency to that found in SCC of the lung. Patients whose tumors showed overexpression of cyclin B1 had a poor event-free survival compared with those lacking this feature (P = 0.04 by Log-rank test). Multivariate analysis of traditional clinical/pathological factors showed that cyclin B1 overexpression was an independent prognostic indicator. CONCLUSIONS: Our study indicates that cyclin B1 is overexpressed in a subset of SCC of the tongue and is associated with a more aggressive biological behavior of the disease. PMID- 11489827 TI - Fluorescence in situ hybridization evaluation of c-erbB-2 gene amplification and chromosomal anomalies in bladder cancer. AB - Oncogene amplification and chromosomal anomalies are found in many solid tumors and are often associated with aggressiveness of cancer. We evaluated the frequency and the role of c-erbB-2 gene amplification, relative increase in c erbB-2 gene copy number, and gain of chromosome 17 in bladder cancer. A total of 29 bladder cancer specimens were examined using fluorescence in situ hybridization (FISH). Dual labeling hybridization with a directly labeled centromere probe for chromosome 17 together with a probe for the c-erbB-2 locus was performed. c-erbB-2 gene amplification was found in 3.4% (1 of 29) of specimens. Relative increase in c-erbB-2 gene copy number was found in 41.4% (12 of 29) of specimens and was significantly associated with tumor grade (P = 0.044 by Fisher's exact test). Gain of chromosome 17 was identified in 65.5% (19 of 29) of specimens and was significantly associated with tumor grade (P = 0.002 by Fisher's exact test) and tumor stage (P = 0.003 by Fisher's exact test). Our results suggest that c-erbB-2 gene amplification, relative increase in c-erbB-2 gene copy number, and gain of chromosome 17 may play important roles in the development and progression of bladder cancers. Moreover, the use of c-erbB-2 amplification, relative increase in c-erbB-2 gene copy number, and gain of chromosome 17 using FISH, together with tumor grade and stage, may provide a more useful clinical indicator in bladder cancer. PMID- 11489828 TI - Expression of X-linked inhibitor of apoptosis as a novel prognostic marker in radically resected non-small cell lung cancer patients. AB - PURPOSE: To assess the pattern of expression and the prognostic value of the inhibitor of apoptosis family member X-linked inhibitor of apoptosis (XIAP; MIHA/ILP-a) in radically resected non-small cell lung cancer patients. EXPERIMENTAL DESIGN: The expression of XIAP and its relationship with overall survival was analyzed by immunohistochemistry on tumors from 144 patients with early-stage non-small cell lung cancer. In addition, the apoptotic and mitotic index, Ki-67, p53, and bcl-2 levels were also assessed. RESULTS: XIAP expression was specific for tumor cells, and the pattern was cytoplasmic. The median expression of XIAP was 20%, and when this value was used as a cutoff point for statistical analyses, 63 of the samples were considered high XIAP-expressing and 81 low XIAP-expressing. Surprisingly, high XIAP-expressing patients had a longer overall survival than the group expressing lower levels (60 versus 24 months of median survival; log rank, P = 0.01). The positive impact of XIAP expression on survival was confirmed by multivariate analysis (P = 0.026). Although no correlation was observed between XIAP expression and the apoptotic index, a significant inverse correlation was observed between XIAP, Ki-67 (P = 0.006), and mitotic index (P = 0.04). CONCLUSIONS: The unexpected inverse correlation of XIAP with proliferation markers and the absence of correlation with apoptotic index, coupled with its role as an independent positive prognostic factor for survival in radically resected NSCLC patients imply a more complex role for XIAP in tumor biology than anticipated by in vitro data. PMID- 11489829 TI - AKT-1, -2, and -3 are expressed in both normal and tumor tissues of the lung, breast, prostate, and colon. AB - PURPOSE: The AKT/PKB kinase controls many of the intracellular processes that are dysregulated in human cancer, including the suppression of apoptosis and anoikis and the induction of cell cycle progression. Three isoforms of AKT have been identified: AKT-1, -2, and -3. Selective up-regulation of AKT-3 RNA expression has been reported in hormone-independent breast and prostate cancer cell lines suggesting that AKT-3 expression may be increased with breast or prostate tumor progression. To determine whether AKT-3 RNA expression is selectively up regulated in human cancers and whether the patterns of AKT RNA expression may change with tumor development, we examined AKT isoform expression by RT-PCR in human cancer cell lines, primary human cancers, and normal human tissues. EXPERIMENTAL DESIGN: AKT-1, -2, and -3 RNA expression was examined by RT-PCR. Because up-regulated AKT-3 expression has been implicated in human breast and prostate cancer progression, we also examined AKT-3 expression levels by semiquantitative RT-PCR using matched normal/tumor first-strand cDNA pairs from colon, breast, prostate, and lung cancers. RESULTS: Our data reveal that the overwhelming majority of both normal and tumor tissues express all three of the AKT isoforms. Moreover, semiquantitative RT-PCR of matched normal/tumor pairs confirmed similar AKT-3 RNA expression levels in both normal and tumor tissue. CONCLUSIONS: Our data show that both normal and tumor tissues express all three of the AKT isoforms and indicate that tumorigenesis does not involve a dramatic shift in the RNA expression patterns of the three AKT isoforms. PMID- 11489830 TI - Death-associated protein 3 (Dap-3) is overexpressed in invasive glioblastoma cells in vivo and in glioma cell lines with induced motility phenotype in vitro. AB - PURPOSE: To discover the genetic determinants of glioma invasion in vivo, we compared the mRNA expression profiles of glioblastoma cells residing at the tumor core versus those at the invasive rim of a human tumor resection. EXPERIMENTAL DESIGN: From a single glioblastoma specimen, 20,000 individual cells from each region (core and invasive rim) were collected by laser capture microdissection and analyzed by mRNA differential display. Differential expression of gene candidates was confirmed by laser capture microdissection and quantitative reverse transcription-PCR in additional glioblastoma multiforme specimens, and the role in migration was further evaluated in glioma cell lines in vitro. RESULTS: Reproducible overexpression the death-associated Protein 3 (Dap-3) mRNA (NM 004632, GenBank; also reported as human ionizing resistance conferring protein mRNA, HSU18321, GenBank) by invasive cells was identified. Although the full-length Dap-3 protein has been described as proapoptotic, the NH(2)-terminal fragment can act in a dominant negative way resulting in protection from programmed cell death. In glioma cell lines T98G and G112 with an induced motility phenotype, Dap-3 was up-regulated at the mRNA and protein level as assessed by quantitative reverse transcription-PCR, cDNA microarray, and Western blot analysis. These cells showed an increased resistance to undergo camptothecin induced apoptosis, which was overcome by effective Dap-3-antisense treatment. Antisense treatment also decreased the migration ability of T98G cells. CONCLUSIONS: Dap-3 is up-regulated in invasive glioblastoma multiforme cells in vivo and in glioma cells with an induced motility phenotype in vitro. When migration is activated, Dap-3 is up-regulated and cells become resistant to apoptosis. These findings suggest that Dap-3 confers apoptosis-resistance when migration behavior is engaged. PMID- 11489831 TI - Synergistic effects of the fenretinide (4-HPR) and anti-CD20 monoclonal antibodies on apoptosis induction of malignant human B cells. AB - Retinoids have been shown to be clinically useful in the biological therapy of certain myeloid and T-cell malignancies, whereas CD20 has proven to be an effective target in B-cell lymphoma immunotherapy. Both retinoic acid derivatives and anti-CD20 monoclonal antibodies have also been shown to induce apoptosis of malignant cells in vitro. Retinoid-induced apoptosis is thought to be mediated by nuclear retinoid receptor binding and transcriptional activation, whereas CD20 ligation appears to initiate transmembrane Ca(2+) influx with resultant programmed cell death. In this report, we evaluate the in vitro effects of N-(4 hydroxyphenyl) retinamide (4-HPR) with and without anti-CD20 antibodies in B-cell lymphoma lines. We demonstrate that 4-HPR inhibits the growth of malignant B cells beyond that of all-trans-retinoic acid and 13-cis-retinoic acid. We also show that this 4-HPR-mediated growth inhibition is attributable to apoptosis, is consistent across a variety of malignant B-cell lines (Ramos, Ramos AW, SU-DHL4, and Raji), peaks at 96 to 144 h, and is attainable with concentrations as low as 2 microM. As with CD20-mediated apoptosis, we show that the final common pathway includes caspase activation that can be blocked by 2-val-Ala-Asp-fluoromethyl ketone (z-VAD), a specific inhibitor of caspase function. Coincubation of a 2 microM concentration of 4-HPR and the anti-CD20 antibodies rituximab and tositumomab exhibited a supra-additive increase in levels of apoptosis induction of 24% (P = 0.009) and 42% (P = 0.0019) relative to expected additive levels of these same agents. These in vitro findings suggest that the potential in vivo synergy of these well-tolerated drugs may augment the previously demonstrated clinical activity of anti-CD20 monoclonal antibodies in the treatment of B-cell malignancies. PMID- 11489832 TI - Strategies to accomplish targeted expression of transgenes in ovarian cancer for molecular therapeutic applications. AB - PURPOSE: The purpose of the study was to determine the capability of the midkine (MK) and cycooxygenase-2 (cox-2) gene promoter regions to function as tumor specific promoters for use in targeted gene therapy of ovarian cancer. EXPERIMENTAL DESIGN: Established and primary ovarian cancer and mesothelial cells were transduced by adenoviral vectors containing a reporter or thymidine kinase gene expressed under the control of the MK, cox-2, or cytomegalovirus (CMV) promoters. SCID or C57BL/6 mice were injected i.p. with these same vectors. In vitro reporter gene expression and cellular cytotoxicity was determined using luciferase and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, respectively. Acute toxicity in vivo was assessed by histological evaluation of harvested tissues. RESULTS: Consistent activation of the MK and cox 2 promoters was noted in all of the ovarian cancer cell lines in addition to primary ovarian cancer cells. In contrast, reduced reporter activity was reported in mesothelial cells transduced with adenoviruses containing the test promoters, which was especially apparent for the cox-2 promoter. Additionally, the cox-2 promoter exhibited significantly lower reporter gene levels in liver and peritoneum than the control promoter in in vivo experiments. Tumor-cell killing induced by Adcox-2 MTK was comparable to that observed with AdCMVTK. However, a clear differential toxicity pattern was observed in favor of animals treated with Adcox-2 MTK when compared with controls. CONCLUSIONS: These data clearly demonstrate that the transcriptional control afforded by the cox-2 promoter is tumor-specific and is able to mitigate associated toxicity in normal tissue while maintaining therapeutic efficacy in the context of an ovarian cancer molecular chemotherapeutic approach. PMID- 11489833 TI - Analysis of cross-resistance of the selective estrogen receptor modulators arzoxifene (LY353381) and LY117018 in tamoxifen-stimulated breast cancer xenografts. AB - PURPOSE: Cross-resistance is the primary issue facing the evaluation of new antiestrogens to treat metastatic breast cancer because they may be tested, initially, in populations of patients that have failed long-term adjuvant tamoxifen (Tam) therapy. EXPERIMENTAL DESIGN: We have tested the benzothiophene derivatives, arzoxifene (Arzox; LY353381) and LY117018 in two models of Tam stimulated tumor growth derived from either MCF-7 (M. M. Gottardis and V. C. Jordan, Cancer Res., 48: 5183-5187, 1988) or T47D (J. MacGregor Schafer et al., Clin. Cancer Res., 6: 4373-4380, 2000) breast cancer cells. RESULTS: Using the MCF-7:Tam model, we found that both Arzox and LY117018 (1.5 mg/day) resulted in tumor growth and, therefore, were partially cross-resistant with Tam. Next, using the T47D:17beta-estradiol (E(2)) model, we compared the antiestrogenic/antitumor properties of Arzox and LY117018 and determined that neither Arzox nor LY117018 caused T47D:E(2) tumor growth after 21 weeks. In addition, we determined that long-term treatment does not result in failure and subsequent development of transplantable Arzox- or LY117018-stimulated tumors. To establish whether Arzox and LY117018 are cross-resistant in T47D:Tam tumors, mice were treated with Arzox or LY117018 (1.5 mg/day), and, again, we found that neither resulted in the growth of transplantable tumors. Lastly, we showed that Arzox and LY117018 were only partially able to compete with postmenopausal E(2) (0.3 cm silastic capsule) in T47D:Tam tumors. However, when T47D:E(2) tumors were treated for 7 days instead of 5 days, both Arzox and LY117018 were more effective. CONCLUSIONS: Arzox is not cross-resistant with Tam in the T47D athymic mouse model but does exhibit cross-resistance in the MCF-7 model. PMID- 11489834 TI - Selective activation of apoptosis program by S-p-bromobenzylglutathione cyclopentyl diester in glyoxalase I-overexpressing human lung cancer cells. AB - PURPOSE: Glyoxalase I (GLO1) is an enzyme that plays a role in the detoxification of methylglyoxal, a side-product of glycolysis. We previously reported that GLO1 was a resistant factor to antitumor agent-induced apoptosis, and that S-p bromobenzylglutathione cyclopentyl diester (BBGC), an effective inhibitor of GLO1, selectively sensitized to etoposide the drug-resistant human leukemia cells that overexpressed GLO1. In this study, we quantitatively measured GLO1 enzyme activity in various human solid tumor cells, and the antiproliferative effect of the GLO1 inhibitor was examined. EXPERIMENTAL DESIGN: BBGC-induced apoptosis was assessed by flow cytometry. To evaluate antitumor activity of BBGC in vivo, we developed human cancer xenografts in nude mice. RESULTS: We found that GLO1 enzyme activity was higher in all of the 38 human cancer cell lines that we examined than in the normal tissue samples. Moreover, GLO1 activity was frequently elevated in human lung carcinoma cells. Positive correlation between cellular GLO1 activity and BBGC sensitivity was observed in the lung cancer cell lines. Human lung cancer NCI-H522 and DMS114 cells, expressing higher GLO1 activity, underwent apoptosis when treated with BBGC, whereas A549 cells, expressing lower activity, did not. BBGC induced the activation of the stress activated protein kinases c-Jun NH(2)-terminal kinase 1 (JNK1) and p38 mitogen activated protein kinase (MAPK), which led to caspase activation in GLO1 overexpressing tumor cells. BBGC significantly inhibited the growth of xenografted DMS114 and human prostate cancer DU-145. CONCLUSIONS: Our present results indicate that GLO1 is a tumor-specific target enzyme especially in human lung carcinoma cells and that the GLO1 inhibitor is a potent chemotherapeutic agent to repress GLO1-overexpressing human tumors. PMID- 11489835 TI - Stable transfection of urokinase-type plasminogen activator antisense construct modulates invasion of human glioblastoma cells. AB - The diffuse and extensive infiltration of malignant gliomas into the surrounding normal brain is believed to rely on modifications of the proteolysis of extracellular matrix components. A key molecule in regulating plasminogen mediated extracellular proteolysis is the urokinase-type plasminogen activator (uPA). To investigate the role of uPA in the invasive process of brain tumors, we stably transfected a human glioblastoma cell line SNB19 with a vector capable of expressing an antisense transcript complementary to the 1020 bases at the 3' end of the uPA cDNA. Parental, vector-, and antisense construct-stably transfected cell lines were analyzed for uPA mRNA transcript by Northern blot analysis, for uPA enzyme activity by zymography, and for uPA protein levels by Western blotting. The levels of uPA mRNA, protein, and enzyme activities were significantly lower in antisense clones than in parental and vector controls. Radioreceptor binding studies demonstrated that uPA receptor levels remained the same in parental, vector-, and antisense-transfected cells. The antisense transfected cells showed a markedly lower level of invasion in the Matrigel invasion assays, and their spheroids failed to invade the fetal rat brain aggregates in the coculture system. Green fluorescent protein (GFP) expressing parental and antisense transfectants was generated for detection in mouse brain tissue without any posttreatment. Intracerebral injection of antisense stable transfectants significantly reduced tumor formation compared with that in controls. Our results suggested that down-regulation of uPA expression may be a feasible approach to reducing the malignancy and invasiveness of glial tumors. PMID- 11489836 TI - Flavopiridol increases sensitization to gemcitabine in human gastrointestinal cancer cell lines and correlates with down-regulation of ribonucleotide reductase M2 subunit. AB - As a single agent, gemcitabine (2',2'-difluorodeoxycytidine) has shown minimal activity against gastrointestinal malignancies with only a modest improvement in survival in patients with pancreatic cancer. Recently, gemcitabine resistance has been associated with the up-regulation of mRNA and protein levels of the ribonucleotide reductase M2 subunit (RR-M2), a rate-limiting enzyme in DNA synthesis that is cell cycle regulated. In this study we show that flavopiridol, a cyclin-dependent kinase inhibitor, enhances the induction of apoptosis by gemcitabine in human pancreatic, gastric, and colon cancer cell lines. As determined by quantitative fluorescence microscopy, flavopiridol enhanced gemcitabine-induced apoptosis 10-15-fold in all of the cell lines tested in a sequence-dependent manner. This was confirmed by poly(ADP-ribose) polymerase cleavage and mitochondrial cytochrome c release. Colony formation assays confirmed the apoptotic rates, showing complete suppression of colony formation only after exposure to sequential treatment of G(24)-->F(24). This is associated with suppression of the RR-M2 protein. This appears to be related to down regulation of E2F-1, a transcription factor that regulates RR-M2 transcription and hypophosphorylation of pRb. The proteasome inhibitor PS-341 could restore the protein levels of E2F-1 in G(24)-->F(24) treatment indicating that E2F-1 down regulation is attributable to its increased degradation via ubiquitin-proteasome pathway. This also resulted in restoration of RR-M2 mRNA and protein. These results indicate that flavopiridol in gemcitabine-treated cells inhibits parts of the machinery necessary for the transcription induction of RR-M2. Thus, combining flavopiridol with gemcitabine may provide an important and novel new means of enhancing the efficacy of gemcitabine in the treatment of gastrointestinal cancers. PMID- 11489837 TI - Combined blockade of protein kinase A and bcl-2 by antisense strategy induces apoptosis and inhibits tumor growth and angiogenesis. AB - Protein kinase A type I (PKAI) plays a key role in neoplastic transformation, conveys mitogenic signals from different sources, and is overexpressed in the majority of human tumors. Inhibition of PKAI by different tools results in cancer cell growth inhibition in vitro and in vivo. We and others have recently shown that a novel class of mixed-backbone oligonucleotides targeting the PKAI subunit RIalpha exhibits improved pharmacokinetic properties and antitumor activity accompanied by increased apoptosis in several human cancer types in vitro and in vivo. The role of bcl-2 in the control of apoptosis has been widely documented, and the inhibition of bcl-2 expression and function may have important therapeutic implications. In fact, oligonucleotides antisense bcl-2 have shown antitumor activity in animal models and have successfully completed early clinical trials. Recent studies have demonstrated a direct role of PKA in the regulation of the bcl-2-dependent apoptotic pathway. Therefore, we have investigated the combined blockade of PKA and bcl-2 by antisense strategy as a potential therapeutic approach. The novel hybrid DNA/RNA mixed-backbone oligonucleotide antisense RIalpha (AS RIalpha) in combination with the antisense bcl-2 (AS bcl-2), cooperatively inhibited bcl-2 expression and soft agar growth and induced apoptosis in different human cancer cell lines. p.o. administration of AS RIalpha in combination with i.p. AS bcl-2 caused a marked antitumor effect and a significant prolongation of survival in nude mice bearing human colon cancer xenografts. Moreover, histochemical analysis of tumor specimens showed inhibition of RIalpha and Ki67 expression, inhibition of angiogenesis, and parallel induction of apoptosis in vivo. The results of our study imply an interaction between the PKA and bcl-2 signaling pathways and, because both antisenses have now entered Phase II trials, provide the rationale to translate this novel therapeutic strategy in a clinical setting. PMID- 11489838 TI - Inhibition of insulin-like growth factor signaling pathways in mammary gland by pure antiestrogen ICI 182,780. AB - The antiestrogens ICI 182,780 (ICI) and tamoxifen are clinically useful in the treatment of estrogen receptor-positive breast tumors. We assessed the in vivo effects of ICI, tamoxifen, and estradiol on the insulin-like growth factor (IGF) signaling pathway in the rat mammary gland. ICI significantly decreased the size of the lobular structures, Ki-67 labeling index, and insulin-like growth factor binding protein (IGFBP)-2 and IGFBP-5 gene expression. Treatment of rats with 1, 1.5, and 2 mg of ICI/kg body weight/week resulted in a 2-, 7-, and 8-fold increase in IGFBP-3 transcripts. High doses of ICI increased mammary IGF-1 gene expression by 2-fold (P < 0.01) but decreased IGF-1R and its autophosphorylation to approximately 30% of the control mammary gland. IRS-1, IRS-2, and c-Raf-1 levels in the ICI-treated mammary glands were approximately 30, 15, and 40% of controls, respectively. Basal phosphorylation of IRS-1, Akt-1, and the p85 subunit of phosphatidylinositol 3-kinase (PI-3K) were low but detectable after ICI treatment. Despite a significant reduction in levels of IGF-1R, IRS-1, and IRS-2 phosphorylation, phospho p42/p44 MAPK levels were only slightly decreased. Tamoxifen-induced growth inhibition was associated with slight stimulation of IGFBP-3 gene expression and reduction in IRS-2 levels. Basal phosphorylation of IGF-1R, IRS-1, and p85 subunit of PI-3K was decreased by tamoxifen. Estradiol induced epithelial cell proliferation was associated with inhibition of IGFBP-3 gene expression, stimulation of IGFBP-2 gene expression, and increases in IGF-1R, IRS-1, IRS-2, and c-Raf-1 levels. Although basal phosphorylation of IGF-1R, IRS 1, IRS-2, Akt-1, and the p85 subunit of PI-3K was significantly increased by estradiol, basal phospho p44/42 MAPK was significantly reduced. The data indicate that in addition to their classic actions, antiestrogens have major effects on IGF signaling pathways. PMID- 11489839 TI - A hammerhead ribozyme suppresses expression of hepatocyte growth factor/scatter factor receptor c-MET and reduces migration and invasiveness of breast cancer cells. AB - PURPOSE: Hepatocyte growth factor/scatter factor (HGF/SF), via its receptor c MET, has been implicated to play a pivotal role in breast cancer development and progression. This study examined a transgene-consisting of a combination of U1snRNA, hammerhead ribozyme, and antisense, designed to inhibit c-met expression and its impact on the migration and in vitro invasion of breast cancer cells. EXPERIMENTAL DESIGN: A hammerhead ribozyme targeting human c-MET was cloned into a modified pZeoU1EcoSpe vector and transfected into breast cancer cells MDA MB 231 and MCF-7 by electroporation. Expression of MET mRNA and protein was determined. Migration and in vitro invasiveness of transfected cells were also analyzed. RESULTS: Breast cancer cells were transfected with the ribozyme containing plasmids. Stable transfectants manifested an almost complete loss of MET mRNA and protein, as shown by reverse transcription-PCR, Northern blotting, and Western blotting, respectively, whereas the wild-type plasmid had no effects. Met-ribozyme transfected cells exhibited reduced migration and in vitro invasiveness through extracellular matrix (Matrigel), compared with the wild-type cells and cells transfected with empty plasmid. CONCLUSIONS: These data show that targeting c-MET by way of a hammerhead ribozyme encoding antisense to c-MET is an effective approach in reducing the invasiveness of breast cancer cells. PMID- 11489840 TI - Optimization for the blockade of epidermal growth factor receptor signaling for therapy of human pancreatic carcinoma. AB - We determined the optimal administration schedule of a novel epidermal growth factor receptor (EGFR) protein tyrosine kinase inhibitor (PKI), PKI 166 (4-(R) phenethylamino-6-(hydroxyl)phenyl-7H-pyrrolo[2.3-d]-pyrimidine), alone or in combination with gemcitabine (administered i.p.) for therapy of L3.6pl human pancreatic carcinoma growing in the pancreas of nude mice. Seven days after orthotopic implantation of L3.6pl cells, the mice received daily oral doses of PKI 166. PKI 166 therapy significantly inhibited phosphorylation of the EGFR without affecting EGFR expression. EGFR phosphorylation was restored 72 h after cessation of therapy. Seven days after orthotopic injection of L3.6pl cells, groups of mice received daily or thrice weekly oral doses of PKI 166 alone or in combination with gemcitabine. Treatment with PKI 166 (daily), PKI 166 (3 times/week), or gemcitabine alone produced a 72%, 69%, or 70% reduction in the volume of pancreatic tumors in mice, respectively. Daily oral PKI 166 or thrice weekly oral PKI 166 in combination with injected gemcitabine produced 97% and 95% decreases in volume of pancreatic cancers and significant inhibition of lymph node and liver metastasis. Daily oral PKI 166 produced a 20% decrease in body weight, whereas treatment 3 times/week did not. Decreased microvessel density, decreased proliferating cell nuclear antigen staining, and increased tumor cell and endothelial cell apoptosis correlated with therapeutic success. Collectively, our results demonstrate that three weekly oral administrations of an EGFR tyrosine kinase inhibitor in combination with gemcitabine are sufficient to significantly inhibit primary and metastatic human pancreatic carcinoma. PMID- 11489841 TI - Marked antitumor activity of a new potent acronycine derivative in orthotopic models of human solid tumors. AB - S 23906-1 is a novel acronycine derivative selected on the basis of its potency in vitro. We investigated the antitumor activity of S 23906-1 against several murine transplantable tumors (C38 colon carcinoma, P388 leukemia, B16 melanoma, and Lewis lung carcinoma) and in orthotopic models of human lung (NCI-H460 and A549), ovarian (IGROV1 and NIH:OVCAR-3), and colorectal cancers (HCT116 and HT 29). Against established C38 colon carcinoma, S 23906-1 administered twice i.v. from 1.56-6.25 mg/kg markedly inhibited tumor growth. Treatment at the optimal dose (6.25 mg/kg) induced tumor regression in all of the mice. Acronycine was 16 fold less potent and only moderately active at the maximum tolerated dose, 100 mg/kg. Against other murine tumors of the former National Cancer Institute panel, S 23906-1 was either only moderately active or totally inactive. When evaluated in human orthotopic models, S 23906-1 given p.o. or i.v. demonstrated a marked antitumor activity against human carcinomas. In the two human lung cancer models, S 23906-1 increased the survival of the animals in a dose-dependent manner and induced treated versus control values of 162% (NCI-H460) and 193% (A549). Vinorelbine was less active, with treated versus control values of 119% and 174%, respectively. A significant survival benefit was also observed against the two i.p. ovarian tumors in which S 23906-1 was as active as paclitaxel, inducing 80% long-term survivors in the NIH:OVCAR-3 model. Lastly, S 23906-1 inhibited the growth of primary HT-29 and HCT116 colon tumors grafted onto the cecum as efficiently as irinotecan and eradicated the formation of lymph node, hepatic, and pulmonary metastases in the aggressive HCT116 model. The novel spectrum of activity of S 23906-1 compared with existing anticancer agents warrants further preclinical investigation. PMID- 11489842 TI - Radiosensitization by gemcitabine in p53 wild-type and mutant MCF-7 breast carcinoma cell lines. AB - The nucleoside analogue 2',2'-difluoro-2'-deoxycytidine (dFdCyd) is a potent radiosensitizer in several solid tumor cell lines. Radiosensitization has correlated with the dFdCyd-mediated decrease in dATP levels and is S-phase specific. Previous studies suggested that a cell line that was unable to progress through S phase after dFdCyd and radiation was not radiosensitized apparently because of the expression of wild-type p53. We have extended these results by using the MCF-7 human breast carcinoma cell line (wild-type p53) and the MCF 7/Adr subline (mutant p53) to determine whether p53 status affected radiosensitization or cell cycle progression after dFdCyd and radiation treatment. Both cell lines were sensitive to nanomolar concentrations of dFdCyd and showed significant radiosensitization, with radiation enhancement ratios of 1.6-1.8 after a 24-h exposure to either the IC(10) or IC(50) for dFdCyd. Nucleotide pool analysis demonstrated a >85% reduction in dATP pools in both cell lines within 8 h after drug addition. Both cell lines accumulated in S phase after a 24-h incubation with dFdCyd. After subsequent irradiation, MCF-7/Adr cells continued to progress through the cell cycle for at least 72 h. MCF-7 cells progressed for at least 24 h, and then exhibited a G(1) block at 48 h after drug and radiation treatment. These results demonstrate that a wild-type p53 cell line can be radiosensitized by dFdCyd, presumably because it was able to deplete dATP levels and progress through the cell cycle for at least 24 h after drug and radiation treatment. PMID- 11489843 TI - The novel tubulin-binding drug BTO-956 inhibits R3230AC mammary carcinoma growth and angiogenesis in Fischer 344 rats. AB - BTO-956 [methyl-3,5-diiodo-4-(4'-methoxyphenoxy)benzoate], a novel tubulin binding drug and thyroid hormone analogue, was originally found to inhibit human carcinoma cell proliferation in vitro and to have potent growth delay activity in human breast and ovarian carcinoma xenografts in nude mice. Here we report that BTO-956 given to Fischer 344 rats also inhibits corneal angiogenesis and the growth and neovascularization of the R3230Ac rat mammary carcinoma tumor implanted in skin-fold window chambers. Hydron pellets containing recombinant human basic fibroblast growth factor (50 ng) and Sucralfate (20 microg) were implanted into surgically created corneal micropockets (day 0). BTO-956 was administrated by oral gavage (500 mg/kg, twice a day for 6 days) on days 1-6 (controls received vehicle alone). On day 7, rats received retrograde infusions of India ink via the thoracic aorta to visualize the corneal vasculature. Digitized images of slide-mounted corneas from control and treated animals were taken with a microscope. For the tumor growth and angiogenesis study, small pieces of R3230Ac tumor from a donor rat were implanted into surgically prepared window chambers (day 0). BTO-956 was given during days 5-11, and images of the tumors and their vasculature were recorded on day 12. No body weight loss was observed in either study. BTO-956 significantly inhibited corneal angiogenesis (by 50-80%), as assessed by measurements of limbal circumference displaying neovascularization, vessel length, vascularized area, and vascular area density. In the window chamber assay, tumors from treated animals were >50% smaller than tumors in control animals. In addition, vascular length densities in peripheral tumor zones were 30% less in treated compared with control animals. Together, these findings demonstrate that BTO-956 can inhibit angiogenesis induced by a growth factor in the rat cornea and in the peripheral area of implanted tumors, where tumor angiogenesis is most active. PMID- 11489844 TI - Keeping signals straight in phosphorelay signal transduction. PMID- 11489845 TI - Transcriptional regulation of the two sterol esterification genes in the yeast Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae transcribes two genes, ARE1 and ARE2, that contribute disproportionately to the esterification of sterols. Are2p is the major enzyme isoform in a wild-type cell growing aerobically. This likely results from a combination of differential transcription initiation and transcript stability. By using ARE1 and ARE2 promoter fusions to lacZ reporters, we demonstrated that transcriptional initiation from the ARE1 promoter is significantly reduced compared to that from the ARE2 promoter. Furthermore, the half-life of the ARE2 mRNA is approximately 12 times as long as that of the ARE1 transcript. We present evidence that the primary role of the minor sterol esterification isoform encoded by ARE1 is to esterify sterol intermediates, whereas the role of the ARE2 enzyme is to esterify ergosterol, the end product of the pathway. Accordingly, the ARE1 promoter is upregulated in strains that accumulate ergosterol precursors. Furthermore, ARE1 and ARE2 are oppositely regulated by heme. Under heme-deficient growth conditions, ARE1 was upregulated fivefold while ARE2 was down-regulated. ARE2 requires the HAP1 transcription factor for optimal expression, and both ARE genes are derepressed in a rox1 (repressor of oxygen) mutant genetic background. We further report that the ARE genes are not subject to end product inhibition; neither ARE1 nor ARE2 transcription is altered in an are mutant background, nor does overexpression of either ARE gene alter the response of the ARE-lacZ reporter constructs. Our observations are consistent with an important physiological role for Are1p during anaerobic growth when heme is limiting and sterol precursors may accumulate. Conversely, Are2p is optimally required during aerobiosis when ergosterol is plentiful. PMID- 11489846 TI - Construction of an in vivo nonsense readthrough assay system and functional analysis of ribosomal proteins S12, S4, and S5 in Bacillus subtilis. AB - To investigate the function of ribosomal proteins and translational factors in Bacillus subtilis, we developed an in vivo assay system to measure the level of nonsense readthrough by utilizing the LacZ-LacI system. Using the in vivo nonsense readthrough assay system which we developed, together with an in vitro poly(U)-directed cell-free translation assay system, we compared the processibility and translational accuracy of mutant ribosomes with those of the wild-type ribosome. Like Escherichia coli mutants, most S12 mutants exhibited lower frequencies of both UGA readthrough and missense error; the only exception was a mutant (in which Lys-56 was changed to Arg) which exhibited a threefold higher frequency of readthrough than the wild-type strain. We also isolated several ribosomal ambiguity (ram) mutants from an S12 mutant. These ram mutants and the S12 mutant mentioned above (in which Lys-56 was changed to Arg) exhibited higher UGA readthrough levels. Thus, the mutation which altered Lys-56 to Arg resulted in a ram phenotype in B. subtilis. The efficacy of our in vivo nonsense readthrough assay system was demonstrated in our investigation of the function of ribosomal proteins and translational factors. PMID- 11489847 TI - Role of DnaB helicase in UV-induced illegitimate recombination in Escherichia coli. AB - To study the involvement of DNA replication in UV-induced illegitimate recombination, we examined the effect of temperature-sensitive dnaB mutations on illegitimate recombination and found that the frequency of illegitimate recombination was reduced by an elongation-deficient mutation, dnaB14, but not by an initiation-deficient mutation, dnaB252. This result indicates that DNA replication is required for UV-induced illegitimate recombination. In addition, the dnaB14 mutation also affected spontaneous or UV-induced illegitimate recombination enhanced by the recQ mutation. Nucleotide sequence analyses of the recombination junctions showed that DnaB-mediated illegitimate recombination is short homology dependent. Previously, Michel et al. (B. Michel, S. Ehrlich, and M. Uzest, EMBO J. 16:430--438, 1997) showed that thermal treatment of the temperature-sensitive dnaB8 mutant induces double-stranded breaks, implying that induction of illegitimate recombination occurs. To explain the discrepancy between the observations, we propose a model for DnaB function, in which the dnaB mutations may exhibit two types of responses, early and late responses, for double-stranded break formation. In the early response, replication forks stall at damaged DNA, resulting in the formation of double-stranded breaks, and the dnaB14 mutation reduces the double-stranded breaks shortly after temperature shift-up. On the other hand, in the late response, the arrested replication forks mediated by the dnaB8 mutation may induce double-stranded breaks after prolonged incubation. PMID- 11489849 TI - Cellobiose uptake in the hyperthermophilic archaeon Pyrococcus furiosus is mediated by an inducible, high-affinity ABC transporter. AB - The hyperthermophilic archaeon Pyrococcus furiosus can utilize different beta glucosides, like cellobiose and laminarin. Cellobiose uptake occurs with high affinity (K(m) = 175 nM) and involves an inducible binding protein-dependent transport system. The cellobiose binding protein (CbtA) was purified from P. furiosus membranes to homogeneity as a 70-kDa glycoprotein. CbtA not only binds cellobiose but also cellotriose, cellotetraose, cellopentaose, laminaribiose, laminaritriose, and sophorose. The cbtA gene was cloned and functionally expressed in Escherichia coli. cbtA belongs to a gene cluster that encodes a transporter that belongs to the Opp family of ABC transporters. PMID- 11489848 TI - A program of Yersinia enterocolitica type III secretion reactions is activated by specific signals. AB - Successful establishment of Yersinia infections requires the type III machinery, a protein transporter that injects virulence factors (Yops) into macrophages. It is reported here that the Yersinia type III pathway responds to environmental signals by transporting proteins to distinct locations. Yersinia enterocolitica cells sense an increase in extracellular amino acids (glutamate, glutamine, aspartate, and asparagine) that results in the activation of the type III pathway. Another signal, provided by serum proteins such as albumin, triggers the secretion of YopD into the extracellular medium. The third signal, a decrease in calcium concentration, appears to be provided by host cells and causes Y. enterocolitica to transport YopE and presumably other virulence factors across the eukaryotic plasma membrane. Mutations in several genes encoding regulatory molecules (lcrG, lcrH, tyeA, yopD, yopN, yscM1, and yscM2) bypass the signal requirement of the type III pathway. Together these results suggest that yersiniae may have evolved distinct secretion reactions in response to environmental signals. PMID- 11489850 TI - The Saccharomyces cerevisiae Isw2p-Itc1p complex represses INO1 expression and maintains cell morphology. AB - In the yeast Saccharomyces cerevisiae, IRE1 encodes a bifunctional protein with transmembrane kinase and endoribonuclease activities. HAC1 encodes a transcription factor which has a basic leucine zipper domain. Both gene products play a crucial role in the unfolded protein response. Mutants in which one of these genes is defective also show the inositol-auxotrophic (Ino(-)) phenotype, but the reason for this has not been clear. To investigate the mechanism underlying the Ino(-) phenotype, we screened a multicopy suppressor gene which can suppress the Ino(-) phenotype of the Delta hac1 strain. We obtained a truncated form of the ITC1 gene that has a defect in its 3' region. Although the truncated form of ITC1 clearly suppressed the Ino(-) phenotype of the Delta hac1 strain, the full-length ITC1 had a moderate effect. The gene products of ITC1 and ISW2 are known to constitute a chromatin-remodeling complex (T. Tsukiyama, J. Palmer, C. C. Landel, J. Shiloach, and C. Wu, Genes Dev. 13:686--697, 1999). Surprisingly, the deletion of either ITC1 or ISW2 in the Delta hac1 strain circumvented the inositol requirement and caused derepression of INO1 even under repression conditions, i.e., in inositol-containing medium. These data indicate that the Isw2p-Itc1p complex usually represses INO1 expression and that overexpression of the truncated form of ITC1 functions in a dominant negative manner in INO1 repression. It is conceivable that the repressor function of this complex is regulated by the C-terminal region of Itc1p. PMID- 11489851 TI - Identification and characterization of fhuD1 and fhuD2, two genes involved in iron-hydroxamate uptake in Staphylococcus aureus. AB - Staphylococcus aureus can utilize several hydroxamate siderophores for growth under iron-restricted conditions. Previous findings have shown that S. aureus possesses a cytoplasmic membrane-associated traffic ATPase that is involved in the specific transport of iron(III)-hydroxamate complexes. In this study, we have identified two additional genes, termed fhuD1 and fhuD2, whose products are involved in this transport process in S. aureus. We have shown that fhuD2 codes for a posttranslationally modified lipoprotein that is anchored in the cytoplasmic membrane, while the deduced amino acid sequence predicts the same for fhuD1. The predicted FhuD1 and FhuD2 proteins share 41.0% identity and 56.4% total similarity with each other, 45.9 and 49.1% total similarity with the FhuD homolog in Bacillus subtilis, and 29.3 and 24.6% total similarity with the periplasmic FhuD protein from Escherichia coli. Insertional inactivation and gene replacement of both genes showed that while FhuD2 is involved in the transport of iron(III) in complex with ferrichrome, ferrioxamine B, aerobactin, and coprogen, FhuD1 shows a more limited substrate range, capable of only iron(III)-ferrichrome and iron(III)-ferrioxamine B transport in S. aureus. Nucleotide sequences present upstream of both fhuD1 and fhuD2 predict the presence of consensus Fur binding sequences. In agreement, transcription of both genes was negatively regulated by exogenous iron levels through the activity of the S. aureus Fur protein. PMID- 11489852 TI - Proteolysis of the Caulobacter McpA chemoreceptor is cell cycle regulated by a ClpX-dependent pathway. AB - Proteolysis is involved in cell differentiation and the progression through the cell cycle in Caulobacter crescentus. We have constitutively expressed the transmembrane chemoreceptor McpA from a multicopy plasmid to demonstrate that McpA degradation is modulated during the cell cycle. The level of McpA protein starts to decrease only when the swarmer cells differentiate into stalked cells. The reduction in McpA protein levels is maintained until the stalked cells develop into predivisional cells, at which point the level returns to that observed in swarmer cells. The cell-cycle-regulated degradation of McpA does not require the last 12 C-terminal amino acids, but it does require three amino acids (AAL) located 15 residues away from the C terminus. The ClpXP protease is essential in C. crescentus for viability, and thus, we tested McpA degradation in xylose conditional mutants. The effect on McpA degradation occurred within two generations from the start of ClpX depletion. The conditional mutants' growth rate was only slightly affected, suggesting that ClpX is directly involved in McpA proteolysis. PMID- 11489853 TI - Characterization of PitA and PitB from Escherichia coli. AB - Escherichia coli contains two major systems for transporting inorganic phosphate (P(i)). The low-affinity P(i) transporter (pitA) is expressed constitutively and is dependent on the proton motive force, while the high-affinity Pst system (pstSCAB) is induced at low external P(i) concentrations by the pho regulon and is an ABC transporter. We isolated a third putative P(i) transport gene, pitB, from E. coli K-12 and present evidence that pitB encodes a functional P(i) transporter that may be repressed at low P(i) levels by the pho regulon. While a pitB(+) cosmid clone allowed growth on medium containing 500 microM P(i), E. coli with wild-type genomic pitB (pitA Delta pstC345 double mutant) was unable to grow under these conditions, making it indistinguishable from a pitA pitB Delta pstC345 triple mutant. The mutation Delta pstC345 constitutively activates the pho regulon, which is normally induced by phosphate starvation. Removal of pho regulation by deleting the phoB-phoR operon allowed the pitB(+) pitA Delta pstC345 strain to utilize P(i), with P(i) uptake rates significantly higher than background levels. In addition, the apparent K(m) of PitB decreased with increased levels of protein expression, suggesting that there is also regulation of the PitB protein. Strain K-10 contains a nonfunctional pitA gene and lacks Pit activity when the Pst system is mutated. The pitA mutation was identified as a single base change, causing an aspartic acid to replace glycine 220. This mutation greatly decreased the amount of PitA protein present in cell membranes, indicating that the aspartic acid substitution disrupts protein structure. PMID- 11489854 TI - Unusual regulatory elements for iron deficiency induction of the idiA gene of Synechococcus elongatus PCC 7942. AB - Expression of a thylakoid membrane-associated protein called IdiA (iron deficiency-induced protein A) is highly elevated and tightly regulated by iron limitation in Synechococcus elongatus PCC 6301 and PCC 7942. Although this protein is not essential for photosystem II (PSII) activity, it plays an important role in protecting the acceptor side of PSII against oxidative damage, especially under iron-limiting growth conditions, by an unknown mechanism. We defined the iron-responsive idiA promoter by using insertional inactivation mutagenesis and reporter gene assays. A 67-bp DNA region was sufficient for full iron deficiency-inducible idiA promoter activity. Within this fragment is a palindromic sequence 4 bp upstream of a putative -35 promoter element, which resembles the binding site of FNR/CAP-type helix-turn-helix transcription factors. The absence of this palindromic sequence or a 3-bp mutation in a putative -10 region eliminated promoter activity completely. A previously identified candidate for a positively acting transcription factor is the IdiB protein, whose gene lies immediately downstream of idiA. IdiB shows strong similarity to helix-turn-helix transcription factors of the FNR/CAP family. A His(6x)-tagged IdiB that was overexpressed in Escherichia coli bound to a 59-bp fragment of the idiA regulatory region that included the palindrome. Although the idiA promoter lacks a consensus binding site for the iron-sensing regulator Fur, we attempted to inactivate fur in order to investigate the potential role of this factor. The resulting merodiploid mutants showed constitutive partial derepression of IdiA expression under iron-sufficient growth conditions. We concluded that IdiB is a specific iron-responsive regulator of idiA and that Fur has an indirect role in influencing idiA expression. PMID- 11489855 TI - Characterizations of highly expressed genes of four fast-growing bacteria. AB - Predicted highly expressed (PHX) genes are characterized for the completely sequenced genomes of the four fast-growing bacteria Escherichia coli, Haemophilus influenzae, Vibrio cholerae, and Bacillus subtilis. Our approach to ascertaining gene expression levels relates to codon usage differences among certain gene classes: the collection of all genes (average gene), the ensemble of ribosomal protein genes, major translation/transcription processing factors, and genes for polypeptides of chaperone/degradation complexes. A gene is predicted highly expressed (PHX) if its codon frequencies are close to those of the ribosomal proteins, major translation/transcription processing factor, and chaperone/degradation standards but strongly deviant from the average gene codon frequencies. PHX genes identified by their codon usage frequencies among prokaryotic genomes commonly include those for ribosomal proteins, major transcription/translation processing factors (several occurring in multiple copies), and major chaperone/degradation proteins. Also PHX genes generally include those encoding enzymes of essential energy metabolism pathways of glycolysis, pyruvate oxidation, and respiration (aerobic and anaerobic), genes of fatty acid biosynthesis, and the principal genes of amino acid and nucleotide biosyntheses. Gene classes generally not PHX include most repair protein genes, virtually all vitamin biosynthesis genes, genes of two-component sensor systems, most regulatory genes, and most genes expressed in stationary phase or during starvation. Members of the set of PHX aminoacyl-tRNA synthetase genes contrast sharply between genomes. There are also subtle differences among the PHX energy metabolism genes between E. coli and B. subtilis, particularly with respect to genes of the tricarboxylic acid cycle. The good agreement of PHX genes of E. coli and B. subtilis with high protein abundances, as assessed by two-dimensional gel determination, is verified. Relationships of PHX genes with stoichiometry, multifunctionality, and operon structures are also examined. The spatial distribution of PHX genes within each genome reveals clusters and significantly long regions without PHX genes. PMID- 11489856 TI - Release of compact nucleoids with characteristic shapes from Escherichia coli. AB - The genomic DNA of bacteria is contained in one or a few compact bodies known as nucleoids. We describe a simple procedure that retains the general shape and compaction of nucleoids from Escherichia coli upon cell lysis and nucleoid release from the cell envelope. The procedure is a modification of that used for the preparation of spermidine nucleoids (nucleoids released in the presence of spermidine) (T. Kornberg, A. Lockwood, and A. Worcel, Proc. Natl. Acad. Sci. USA 71:3189--3193, 1974). Polylysine is added to prevent the normal decompaction of nucleoids which occurs upon cell lysis. Nucleoids retained their characteristic shapes in lysates of exponential-phase cells or in lysates of cells treated with chloramphenicol or nalidixate to alter nucleoid morphology. The notably unstable nucleoids of rifampin-treated cells were obtained in compact, stable form in such lysates. Nucleoids released in the presence of polylysine were easily processed and provided well-defined DNA fluorescence and phase-contrast images. Uniform populations of nucleoids retaining characteristic shapes could be isolated after formaldehyde fixation and heating with sodium dodecyl sulfate. PMID- 11489857 TI - Extracellular synthesis, specific recognition, and intracellular degradation of cyclomaltodextrins by the hyperthermophilic archaeon Thermococcus sp. strain B1001. AB - A unique extracellular and thermostable cyclomaltodextrin glucanotransferase (CGTase) from the hyperthermophilic archaeon Thermococcus sp. strain B1001 produces predominantly (>85%) alpha-cyclomaltodextrin (alpha-CD) from starch (Y. Tachibana, et al., Appl. Environ. Microbiol. 65:1991--1997, 1999). Nucleotide sequencing of the CGTase gene (cgtA) and its flanking region was performed, and a cluster of five genes was found, including a gene homolog encoding a cyclomaltodextrinase (CDase) involved in the degradation of CDs (cgtB), the gene encoding CGTase (cgtA), a gene homolog for a CD-binding protein (CBP) (cgtC), and a putative CBP-dependent ABC transporter involved in uptake of CDs (cgtDE). The CDase was expressed in Escherichia coli and purified. The optimum pH and temperature for CD hydrolysis were 5.5 and 95 degrees C, respectively. The molecular weight of the recombinant enzyme was estimated to be 79,000. The CDase hydrolyzed beta-CD most efficiently among other CDs. Maltose and pullulan were not utilized as substrates. Linear maltodextrins with a small glucose unit were very slowly hydrolyzed, and starch was hydrolyzed more slowly. Analysis by thin layer chromatography revealed that glucose and maltose were produced as end products. The purified recombinant CBP bound to maltose as well as to alpha-CD. However, the CBP exhibited higher thermostability in the presence of alpha-CD. These results suggested that strain B1001 possesses a unique metabolic pathway that includes extracellular synthesis, transmembrane uptake, and intracellular degradation of CDs in starch utilization. Potential advantages of this starch metabolic pathway via CDs are discussed. PMID- 11489858 TI - Halohydrin dehalogenases are structurally and mechanistically related to short chain dehydrogenases/reductases. AB - Halohydrin dehalogenases, also known as haloalcohol dehalogenases or halohydrin hydrogen-halide lyases, catalyze the nucleophilic displacement of a halogen by a vicinal hydroxyl function in halohydrins to yield epoxides. Three novel bacterial genes encoding halohydrin dehalogenases were cloned and expressed in Escherichia coli, and the enzymes were shown to display remarkable differences in substrate specificity. The halohydrin dehalogenase of Agrobacterium radiobacter strain AD1, designated HheC, was purified to homogeneity. The k(cat) and K(m) values of this 28-kDa protein with 1,3-dichloro-2-propanol were 37 s(-1) and 0.010 mM, respectively. A sequence homology search as well as secondary and tertiary structure predictions indicated that the halohydrin dehalogenases are structurally similar to proteins belonging to the family of short-chain dehydrogenases/reductases (SDRs). Moreover, catalytically important serine and tyrosine residues that are highly conserved in the SDR family are also present in HheC and other halohydrin dehalogenases. The third essential catalytic residue in the SDR family, a lysine, is replaced by an arginine in halohydrin dehalogenases. A site-directed mutagenesis study, with HheC as a model enzyme, supports a mechanism for halohydrin dehalogenases in which the conserved Tyr145 acts as a catalytic base and Ser132 is involved in substrate binding. The primary role of Arg149 may be lowering of the pK(a) of Tyr145, which abstracts a proton from the substrate hydroxyl group to increase its nucleophilicity for displacement of the neighboring halide. The proposed mechanism is fundamentally different from that of the well-studied hydrolytic dehalogenases, since it does not involve a covalent enzyme-substrate intermediate. PMID- 11489859 TI - Functional and evolutionary relationship between arginine biosynthesis and prokaryotic lysine biosynthesis through alpha-aminoadipate. AB - Our previous studies revealed that lysine is synthesized through alpha aminoadipate in an extremely thermophilic bacterium, Thermus thermophilus HB27. Sequence analysis of a gene cluster involved in the lysine biosynthesis of this microorganism suggested that the conversion from alpha-aminoadipate to lysine proceeds in a way similar to that of arginine biosynthesis. In the present study, we cloned an argD homolog of T. thermophilus HB27 which was not included in the previously cloned lysine biosynthetic gene cluster and determined the nucleotide sequence. A knockout of the argD-like gene, now termed lysJ, in T. thermophilus HB27 showed that this gene is essential for lysine biosynthesis in this bacterium. The lysJ gene was cloned into a plasmid and overexpressed in Escherichia coli, and the LysJ protein was purified to homogeneity. When the catalytic activity of LysJ was analyzed in a reverse reaction in the putative pathway, LysJ was found to transfer the epsilon-amino group of N(2)-acetyllysine, a putative intermediate in lysine biosynthesis, to 2-oxoglutarate. When N(2) acetylornithine, a substrate for arginine biosynthesis, was used as the substrate for the reaction, LysJ transferred the delta-amino group of N(2)-acetylornithine to 2-oxoglutarate 16 times more efficiently than when N(2)-acetyllysine was the amino donor. All these results suggest that lysine biosynthesis in T. thermophilus HB27 is functionally and evolutionarily related to arginine biosynthesis. PMID- 11489860 TI - Genetic and structural organization of the aminophenol catabolic operon and its implication for evolutionary process. AB - The aminophenol (AP) catabolic operon in Pseudomonas putida HS12 mineralizing nitrobenzene was found to contain all the enzymes responsible for the conversion of AP to pyruvate and acetyl coenzyme A via extradiol meta cleavage of 2 aminophenol. The sequence and functional analyses of the corresponding genes of the operon revealed that the AP catabolic operon consists of one regulatory gene, nbzR, and the following nine structural genes, nbzJCaCbDGFEIH, which encode catabolic enzymes. The NbzR protein, which is divergently transcribed with respect to the structural genes, possesses a leucine zipper motif and a MarR homologous domain. It was also found that NbzR functions as a repressor for the AP catabolic operon through binding to the promoter region of the gene cluster in its dimeric form. A comparative study of the AP catabolic operon with other meta cleavage operons led us to suggest that the regulatory unit (nbzR) was derived from the MarR family and that the structural unit (nbzJCaCbDGFEIH) has evolved from the ancestral meta cleavage gene cluster. It is also proposed that these two functional units assembled through a modular type gene transfer and then have evolved divergently to acquire specialized substrate specificities (NbzCaCb and NbzD) and catalytic function (NbzE), resulting in the creation of the AP catabolic operon. The evolutionary process of the AP operon suggests how bacteria have efficiently acquired genetic diversity and expanded their metabolic capabilities by modular type gene transfer. PMID- 11489861 TI - LcrG-LcrV interaction is required for control of Yops secretion in Yersinia pestis. AB - Yersinia pestis expresses a set of plasmid-encoded virulence proteins called Yops and LcrV that are secreted and translocated into eukaryotic cells by a type III secretion system. LcrV is a multifunctional protein with antihost and positive regulatory effects on Yops secretion that forms a stable complex with a negative regulatory protein, LcrG. LcrG has been proposed to block the secretion apparatus (Ysc) from the cytoplasmic face of the inner membrane under nonpermissive conditions for Yops secretion, when levels of LcrV in the cell are low. A model has been proposed to describe secretion control based on the relative levels of LcrG and LcrV in the bacterial cytoplasm. This model proposes that under secretion-permissive conditions, levels of LcrV are increased relative to levels of LcrG, so that the excess LcrV titrates LcrG away from the Ysc, allowing secretion of Yops to occur. To further test this model, a mutant LcrG protein that could no longer interact with LcrV was created. Expression of this LcrG variant blocked secretion of Yops and LcrV under secretion permissive conditions in vitro and in a tissue culture model. These results agree with the previously described secretion-blocking activity of LcrG and demonstrate that the interaction of LcrV with LcrG is necessary for controlling Yops secretion. PMID- 11489862 TI - Differential regulation of ftsZ transcription during septation of Streptomyces griseus. AB - Streptomyces has been known to form two types of septa. The data in this research demonstrated that Streptomyces griseus forms another type of septum near the base of sporogenic hyphae (basal septum). To understand the regulation of the septation machinery in S. griseus, we investigated the expression of the ftsZ gene. S1 nuclease protection assays revealed that four ftsZ transcripts were differentially expressed during morphological differentiation. The vegetative transcript (emanating from P(veg)) is present at a moderate level during vegetative growth, but is switched off within the first 2 h of sporulation. Two sporulation-specific transcripts predominantly accumulated, and the levels increased by approximately fivefold together shortly before sporulation septa begin to form. Consistently, the sporulation-specific transcripts were expressed much earlier and more abundantly in a group of nonsporulating mutants that form their sporulation septa prematurely. Promoter-probe studies with two different reporter systems confirmed the activities of the putative promoters identified from the 5' end point of the transcripts. The levels and expression timing of promoter activities were consistent with the results of nuclease protection assays. The aseptate phenotype of the P(spo) mutant indicated that the increased transcription from P(spo) is required for sporulation septation, but not for vegetative or basal septum formation. PMID- 11489863 TI - Insertional mutagenesis in the n-alkane-assimilating yeast Yarrowia lipolytica: generation of tagged mutations in genes involved in hydrophobic substrate utilization. AB - Tagged mutants affected in the degradation of hydrophobic compounds (HC) were generated by insertion of a zeta-URA3 mutagenesis cassette (MTC) into the genome of a zeta-free and ura3 deletion-containing strain of Yarrowia lipolytica. MTC integration occurred predominantly at random by nonhomologous recombination. A total of 8,600 Ura(+) transformants were tested by replica plating for (i) growth on minimal media with alkanes of different chain lengths (decane, dodecane, and hexadecane), oleic acid, tributyrin, or ethanol as the C source and (ii) colonial defects on different glucose-containing media (YPD, YNBD, and YNBcas). A total of 257 mutants were obtained, of which about 70 were affected in HC degradation, representing different types of non-alkane-utilizing (Alk(-)) mutants (phenotypic classes alkA to alkE) and tributyrin degradation mutants. Among Alk(-) mutants, growth defects depending on the alkane chain length were observed (alkAa to alkAc). Furthermore, mutants defective in yeast-hypha transition and ethanol utilization and selected auxotrophic mutants were isolated. Flanking borders of the integrated MTC were sequenced to identify the disrupted genes. Sequence analysis indicated that the MTC was integrated in the LEU1 locus in N083, a leucine-auxotrophic mutant, in the isocitrate dehydrogenase gene of N156 (alkE leaky), in the thioredoxin reductase gene in N040 (alkAc), and in a peroxine gene (PEX14) in N078 (alkD). This indicates that MTC integration is a powerful tool for generating and analyzing tagged mutants in Y. lipolytica. PMID- 11489864 TI - Regulation of the glv operon in Bacillus subtilis: YfiA (GlvR) is a positive regulator of the operon that is repressed through CcpA and cre. AB - Maltose metabolism and the regulation of the glv operon of Bacillus subtilis, comprising three genes, glvA (6-phospho-alpha-glucosidase), yfiA (now designated glvR), and glvC (EIICB transport protein), were investigated. Maltose dissimilation was dependent primarily upon the glv operon, and insertional inactivation of either glvA, glvR, or glvC markedly inhibited growth on the disaccharide. A second system (MalL) contributed to a minor extent to maltose metabolism. Northern blotting revealed two transcripts corresponding to a monocistronic mRNA of glvA and a polycistronic mRNA of glvA-glvR-glvC. Primer extension analysis showed that both transcripts started at the same base (G) located 26 bp upstream of the 5' end of glvA. When glvR was placed under control of the spac promoter, expression of the glv operon was dependent upon the presence of isopropyl-beta-D-thiogalactopyranoside (IPTG). In regulatory studies, the promoter sequence of the glv operon was fused to lacZ and inserted into the amyE locus, and the resultant strain (AMGLV) was then transformed with a citrate controlled glvR plasmid, pHYCM2VR. When cultured in Difco sporulation medium containing citrate, this transformant [AMGLV(pHYCM2VR)] expressed LacZ activity, but synthesis of LacZ was repressed by glucose. In an isogenic strain, [AMGLVCR(pHYCM2VR)], except for a mutation in the sequence of a catabolite responsive element (cre), LacZ activity was expressed in the presence of citrate and glucose. Insertion of a citrate-controlled glvR plasmid at the amyE locus of ccpA(+) and ccpA mutant organisms yielded strains AMCMVR and AMCMVRCC, respectively. In the presence of both glucose and citrate, AMCMVR failed to express the glv operon, whereas under the same conditions high-level expression of both mRNA transcripts was found in strain AMCMVRCC. Collectively, our findings suggest that GlvR (the product of the glvR gene) is a positive regulator of the glv operon and that glucose exerts its effect via catabolite repression requiring both CcpA and cre. PMID- 11489865 TI - Identification of the UDP-MurNAc-pentapeptide:L-alanine ligase for synthesis of branched peptidoglycan precursors in Enterococcus faecalis. AB - Many species of gram-positive bacteria produce branched peptidoglycan precursors resulting from the transfer of various L-amino acids or glycine from amino acyl tRNA to the epsilon-amino group of L-lysine. The UDP-MurNAc-pentapeptide:L alanine ligase and alanyl-tRNA synthetase genes from Enterococcus faecalis were identified, cloned, and overexpressed in Escherichia coli. The purified enzymes were necessary and sufficient for tRNA-dependent addition of L-alanine to UDP MurNAc-pentapeptide in vitro. The ligase belonged to the Fem family of proteins, which were initially identified genetically as factors essential for methicillin resistance in Staphylococcus aureus. PMID- 11489866 TI - Role of ptsO in carbon-mediated inhibition of the Pu promoter belonging to the pWW0 Pseudomonas putida plasmid. AB - An investigation was made into the role of the ptsO gene in carbon source inhibition of the Pu promoter belonging to the Pseudomonas putida upper TOL (toluene degradation) operon. ptsO is coexpressed with ptsN, the loss of which is known to render Pu unresponsive to glucose. Both ptsN and ptsO, coding for the phosphoenolpyruvate:sugar phosphotransferase system (PTS) family proteins IIA(Ntr) and NPr, respectively, have been mapped adjacent to the rpoN gene of P. putida. The roles of these two genes in the responses of Pu to glucose were monitored by lacZ reporter technology with a P. putida strain engineered with all regulatory elements in monocopy gene dosage. In cells lacking ptsO, Pu activity seemed to be inhibited even in the absence of glucose. A functional relationship with ptsN was revealed by the phenotype of a double ptsN ptsO mutant that was equivalent to the phenotype of a mutant with a single ptsN disruption. Moreover, phosphorylation of the product of ptsO seemed to be required for C inhibition of Pu, since an H15A change in the NPr sequence that prevents phosphorylation of this conserved amino acid residue did not restore the wild-type phenotype. A genomic search for proteins able to phosphorylate ptsO revealed the presence of two open reading frames, designated ptsP and mtp, with the potential to encode PTS type I enzymes in P. putida. However, neither an insertion in ptsP nor an insertion in mtp resulted in a detectable change in inhibition of Pu by glucose. These results indicate that some PTS proteins have regulatory functions in P. putida that are independent of their recognized role in sugar transport in other bacteria. PMID- 11489867 TI - Two membrane-associated NiFeS-carbon monoxide dehydrogenases from the anaerobic carbon-monoxide-utilizing eubacterium Carboxydothermus hydrogenoformans. AB - Two monofunctional NiFeS carbon monoxide (CO) dehydrogenases, designated CODH I and CODH II, were purified to homogeneity from the anaerobic CO-utilizing eubacterium Carboxydothermus hydrogenoformans. Both enzymes differ in their subunit molecular masses, N-terminal sequences, peptide maps, and immunological reactivities. Immunogold labeling of ultrathin sections revealed both CODHs in association with the inner aspect of the cytoplasmic membrane. Both enzymes catalyze the reaction CO + H(2)O --> CO(2) + 2 e(-) + 2 H(+). Oxidized viologen dyes are effective electron acceptors. The specific enzyme activities were 15,756 (CODH I) and 13,828 (CODH II) micromol of CO oxidized min(-1) mg(-1) of protein (methyl viologen, pH 8.0, 70 degrees C). The two enzymes oxidize CO very efficiently, as indicated by k(cat)/K(m) values at 70 degrees C of 1.3. 10(9) M( 1) CO s(-1) (CODH I) and 1.7. 10(9) M(-1) CO s(-1) (CODH II). The apparent K(m) values at pH 8.0 and 70 degrees C are 30 and 18 microM CO for CODH I and CODH II, respectively. Acetyl coenzyme A synthase activity is not associated with the enzymes. CODH I (125 kDa, 62.5-kDa subunit) and CODH II (129 kDa, 64.5-kDa subunit) are homodimers containing 1.3 to 1.4 and 1.7 atoms of Ni, 20 to 22 and 20 to 24 atoms of Fe, and 22 and 19 atoms of acid-labile sulfur, respectively. Electron paramagnetic resonance (EPR) spectroscopy revealed signals indicative of [4Fe-4S] clusters. Ni was EPR silent under any conditions tested. It is proposed that CODH I is involved in energy generation and that CODH II serves in anabolic functions. PMID- 11489868 TI - Diversity of Streptococcus salivarius ptsH mutants that can be isolated in the presence of 2-deoxyglucose and galactose and characterization of two mutants synthesizing reduced levels of HPr, a phosphocarrier of the phosphoenolpyruvate:sugar phosphotransferase system. AB - In streptococci, HPr, a phosphocarrier of the phosphoenolpyruvate:sugar phosphotransferase transport system (PTS), undergoes multiple posttranslational chemical modifications resulting in the formation of HPr(His approximately P), HPr(Ser-P), and HPr(Ser-P)(His approximately P), whose cellular concentrations vary with growth conditions. Distinct physiological functions are associated with specific forms of HPr. We do not know, however, the cellular thresholds below which these forms become unable to fulfill their functions and to what extent modifications in the cellular concentrations of the different forms of HPr modify cellular physiology. In this study, we present a glimpse of the diversity of Streptococcus salivarius ptsH mutants that can be isolated by positive selection on a solid medium containing 2-deoxyglucose and galactose and identify 13 amino acids that are essential for HPr to properly accomplish its physiological functions. We also report the characterization of two S. salivarius mutants that produced approximately two- and threefoldless HPr and enzyme I (EI) respectively. The data indicated that (i) a reduction in the synthesis of HPr due to a mutation in the Shine-Dalgarno sequence of ptsH reduced ptsI expression; (ii) a threefold reduction in EI and HPr cellular levels did not affect PTS transport capacity; (iii) a twofold reduction in HPr synthesis was sufficient to reduce the rate at which cells metabolized PTS sugars, increase generation times on PTS sugars and to a lesser extent on non-PTS sugars, and impede the exclusion of non-PTS sugars by PTS sugars; (iv) a threefold reduction in HPr synthesis caused a strong derepression of the genes coding for alpha-galactosidase, beta-galactosidase, and galactokinase when the cells were grown at the expense of a PTS sugar but did not affect the synthesis of alpha-galactosidase when cells were grown at the expense of lactose, a noninducing non-PTS sugar; and (v) no correlation was found between the magnitude of enzyme derepression and the cellular levels of HPr(Ser-P). PMID- 11489869 TI - Roles of FrxA and RdxA nitroreductases of Helicobacter pylori in susceptibility and resistance to metronidazole. AB - The relative importance of the frxA and rdxA nitroreductase genes of Helicobacter pylori in metronidazole (MTZ) susceptibility and resistance has been controversial. Jeong et al. (J. Bacteriol. 182:5082--5090, 2000) had interpreted that Mtz(s) H. pylori were of two types: type I, requiring only inactivation of rdxA to became resistant, and type II, requiring inactivation of both rdxA and frxA to become resistant; frxA inactivation by itself was not sufficient to confer resistance. In contrast, Kwon et al. (Antimicrob. Agents Chemother. 44:2133--2142, 2000) had interpreted that resistance resulted from inactivation either of frxA or rdxA. These two interpretations were tested here. Resistance was defined as efficient colony formation by single cells from diluted cultures rather than as growth responses of more dense inocula on MTZ-containing medium. Tests of three of Kwon's Mtz(s) strains showed that each was type II, requiring inactivation of both rdxA and frxA to become resistant. In additional tests, derivatives of frxA mutant strains recovered from MTZ-containing medium were found to contain new mutations in rdxA, and frxA inactivation slowed MTZ-induced killing of Mtz(s) strains. Northern blot analyses indicated that frxA mRNA, and perhaps also rdxA mRNA, were more abundant in type II than in type I strains. We conclude that development of MTZ resistance in H. pylori requires inactivation of rdxA alone or of both rdxA and frxA, depending on bacterial genotype, but rarely, if ever, inactivation of frxA alone, and that H. pylori strains differ in regulation of nitroreductase gene expression. We suggest that such regulatory differences may be significant functionally during human infection. PMID- 11489870 TI - The molecular weight distribution of succinoglycan produced by Sinorhizobium meliloti is influenced by specific tyrosine phosphorylation and ATPase activity of the cytoplasmic domain of the ExoP protein. AB - It is thought that in the gram-negative soil bacterium Sinorhizobium meliloti the protein ExoP is involved in biosynthesis of the acidic exopolysaccharide succinoglycan (EPS I). The amounts and compositions of EPS I produced by mutants expressing ExoP proteins characterized by specific amino acid substitutions in the C-terminal cytoplasmic domain were analyzed. The cytoplasmic domain of the ExoP protein was shown to have ATPase activity. Mutations in the highly conserved Walker A ATP-binding motif prevented ATPase activity of the ExoP protein. Phenotypically, these mutations resulted in much lower levels of succinoglycan which consisted only of monomers of the octasaccharide repeating unit. The ExoP protein has similarities to proteins with autophosphorylating protein tyrosine kinase activity. We found that ExoP was phosphorylated on tyrosine and that site directed mutagenesis of specific tyrosine residues in the cytoplasmic domain of ExoP resulted in an altered ratio of low-molecular-weight succinoglycan to high molecular-weight succinoglycan. PMID- 11489871 TI - Influence of a functional sigB operon on the global regulators sar and agr in Staphylococcus aureus. AB - The growth phase-dependent activity profile of the alternate transcription factor sigma(B) and its effects on the expression of sar and agr were examined in three different Staphylococcus aureus strains by Northern blot analyses and by the use of reporter gene fusion experiments. Significant sigma(B) activity was detectable only in the clinical isolates MSSA1112 and Newman, carrying the wild-type rsbU allele, but not in the NCTC8325 derivative BB255, which is defective in rsbU. sigma(B) activity peaked in the late exponential phase and diminished towards the stationary phase when bacteria were grown in Luria-Bertani medium. Transcriptional analysis and a sarP1-sarP2-sarP3 (sarP1-P2-P3)-driven firefly luciferase (luc+) reporter gene fusion demonstrated a strong sigma(B) activity- and growth phase-dependent increase in sar expression that was totally absent in either rsbU or Delta rsbUVWsigB mutants. In contrast, expression of the agr locus, as measured by RNAIII levels and by an hldp::luc+ fusion, was found to be higher in the absence of sigma(B) activity, such as in rsbU or Delta rsbUVWsigB mutants, than in wild-type strains. Overexpression of sigma(B) in BB255 derivatives resulted in a clear increase in sarP1-P2-P3::luc+ expression as well as a strong decrease in hldp::luc+ expression. The data presented here suggest that sigma(B) increases sar expression while simultaneously reducing the RNAIII level in a growth phase-dependent manner. PMID- 11489872 TI - Cytological evidence for association of the ends of the linear chromosome in Streptomyces coelicolor. AB - The chromosome of the filamentous bacterium Streptomyces coelicolor is linear, but the genetic map is circular. We present cytological evidence based on the use of fluorescence in situ hybridization showing that the ends of the chromosome frequently colocalize, in agreement with the idea that the ends are held together, effectively forming a circular chromosome. These observations provide a possible explanation for how a linear bacterial chromosome can exhibit a circular genetic map. PMID- 11489873 TI - Quorum sensing is a global regulatory mechanism in enterohemorrhagic Escherichia coli O157:H7. AB - Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is responsible for outbreaks of bloody diarrhea and hemolytic-uremic syndrome in many countries. EHEC virulence mechanisms include the production of Shiga toxins (Stx) and formation of attaching and effacing (AE) lesions on intestinal epithelial cells. We recently reported that genes involved in the formation of the AE lesion were regulated by quorum sensing through autoinducer-2, which is synthesized by the product of the luxS gene. In this study we hybridized an E. coli gene array with cDNA synthesized from RNA that was extracted from EHEC strain 86-24 and its isogenic luxS mutant. We observed that 404 genes were regulated by luxS at least fivefold, which comprises approximately 10% of the array genes; 235 of these genes were up regulated and 169 were down-regulated in the wild-type strain compared to in the luxS mutant. Down-regulated genes included several involved in cell division, as well as ribosomal and tRNA genes. Consistent with this pattern of gene expression, the luxS mutant grows faster than the wild-type strain (generation times of 37.5 and 60 min, respectively, in Dulbecco modified Eagle medium). Up regulated genes included several involved in the expression and assembly of flagella, motility, and chemotaxis. Using operon::lacZ fusions to class I, II, and III flagellar genes, we were able to confirm this transcriptional regulation. We also observed fewer flagella by Western blotting and electron microscopy and decreased motility halos in semisolid agar in the luxS mutant. The average swimming speeds for the wild-type strain and the luxS mutant are 12.5 and 6.6 microm/s, respectively. We also observed an increase in the production of Stx due to quorum sensing. Genes encoding Stx, which are transcribed along with lambda like phage genes, are induced by an SOS response, and genes involved in the SOS response were also regulated by quorum sensing. These results indicate that quorum sensing is a global regulatory mechanism for basic physiological functions of E. coli as well as for virulence factors. PMID- 11489875 TI - Methanococcus jannaschii generates L-proline by cyclization of L-ornithine. AB - Cell extracts of Methanococcus jannaschii have been shown to readily convert L ornithine to L-proline. This cyclization reaction proceeds with the loss of only the C-2 nitrogen, as has been documented for ornithine cyclodeaminase (EC 4.3.1.12). Since no gene homologous to that coding for ornithine cyclodeaminase is present in the genome of M. jannaschii, these results indicate that proline biosynthesis in M. jannaschii is accomplished by a previously unrecognized enzyme. PMID- 11489874 TI - Regulation and mechanism of action of the small heat shock protein from the hyperthermophilic archaeon Pyrococcus furiosus. AB - The small heat shock protein (sHSP) from the hyperthermophile Pyrococcus furiosus was specifically induced at the level of transcription by heat shock at 105 degrees C. The gene encoding this protein was cloned and overexpressed in Escherichia coli. The recombinant sHSP prevented the majority of E. coli proteins from aggregating in vitro for up to 40 min at 105 degrees C. The sHSP also prevented bovine glutamate dehydrogenase from aggregating at 56 degrees C. Survivability of E. coli overexpressing the sHSP was enhanced approximately sixfold during exposure to 50 degrees C for 2 h compared with the control culture, which did not express the sHSP. Apparently, the sHSP confers a survival advantage on mesophilic bacteria by preventing protein aggregation at supraoptimal temperatures. PMID- 11489876 TI - Lambda excision revisited: testing a model for synapsis of prophage ends. AB - Excision of lambda prophage was reexamined to test a model for prophage end synapsis. The model proposes that, during in situ prophage replication, following induction, the diverging replication forks are held together. Consequently, prophage DNA is spooled through the replication machinery, drawing the prophage ends together and facilitating synapsis. The model predicts that excision will be slowed if in situ lambda replication is inhibited, and the predicted low rate of excision of a nonreplicating prophage was observed after thermoinduction. However, excision was rapid if additional Int protein was supplied or if the temperature was reduced after induction, showing that (i) Int is partially thermosensitive for excision at 42 degrees C and (ii) in situ replication is not required for rapid excision, a finding that is inconsistent with the model. PMID- 11489877 TI - Dehalogenation of dichloromethane by dichloromethane dehalogenase/glutathione S transferase leads to formation of DNA adducts. AB - Formation of DNA adducts following conversion of dichloromethane by bacterial dichloromethane dehalogenase/glutathione S-transferase was demonstrated. Adducts included dichloromethane carbon and glutathione sulfur atoms. A reaction with DNA occurred preferentially at guanine bases. Increased DNA degradation in a polA mutant of Methylobacterium dichloromethanicum DM4 grown with dichloromethane confirmed the genotoxicity associated with dichloromethane degradation, suggesting an important role of DNA repair in the metabolism of halogenated, DNA alkylating compounds by bacteria. PMID- 11489878 TI - Steroidogenic acute regulatory protein binds cholesterol and modulates mitochondrial membrane sterol domain dynamics. AB - The steroidogenic acute regulatory protein (StAR) mediates the rate-limiting step of steroidogenesis, delivery of cholesterol to the inner mitochondrial membrane. However, the mechanism whereby cholesterol translocation is accomplished has not been resolved. Recombinant StAR proteins lacking the first N-terminal 62 amino acids comprising the mitochondrial-targeting sequence were used to determine if StAR binds cholesterol and alters mitochondrial membrane cholesterol domains to enhance sterol transfer. First, a fluorescent NBD-cholesterol binding assay revealed 2 sterol binding sites (K(d) values near 32 nm), whereas the inactive A218V N-62 StAR mutant had only a single binding site with 8-fold lower affinity. Second, NBD-cholesterol spectral shifts and fluorescence resonance energy transfer from StAR Trp residues to NBD-cholesterol showed (i) close molecular interaction between these molecules (R(2/3) = 33 A) and (ii) sensitized NBD cholesterol emission from only one of the two sterol binding sites. Third, circular dichroism showed that cholesterol binding induced a change in StAR secondary structure. Fourth, a fluorescent sterol transfer assay that did not require separation of donor and acceptor mitochondrial membranes demonstrated that StAR enhanced mitochondrial sterol transfer as much as 100-fold and induced/increased the formation of rapidly transferable cholesterol domains in isolated mitochondrial membranes. StAR was 67-fold more effective in transferring cholesterol from mitochondria of steroidogenic MA-10 cells than from human fibroblast mitochondria. In contrast, sterol carrier protein-2 (SCP-2) was only 2.2-fold more effective in mediating sterol transfer from steroidogenic cell mitochondria. Taken together these data showed that StAR is a cholesterol-binding protein, preferentially enhances sterol transfer from steroidogenic cell mitochondria, and interacts with mitochondrial membranes to alter their sterol domain structure and dynamics. PMID- 11489879 TI - Mutations in both sides of the photosystem I reaction center identify the phylloquinone observed by electron paramagnetic resonance spectroscopy. AB - The core of photosystem I (PS1) is composed of the two related integral membrane polypeptides, PsaA and PsaB, which bind two symmetrical branches of cofactors, each consisting of two chlorophylls and a phylloquinone, that potentially link the primary electron donor and the tertiary acceptor. In an effort to identify amino acid residues near the phylloquinone binding sites, all tryptophans and histidines that are conserved between PsaA and PsaB in the region of the 10th and 11th transmembrane alpha-helices were mutated in Chlamydomonas reinhardtii. The mutant PS1 reaction centers appear to assemble normally and possess photochemical activity. An electron paramagnetic resonance (EPR) signal attributed to the phylloquinone anion radical (A(1)(-)) can be observed either transiently or after illumination of reaction centers with pre-reduced iron-sulfur clusters. Mutation of PsaA-Trp(693) to Phe resulted in an inability to photo-accumulate A(1)(-), whereas mutation of the analogous tryptophan in PsaB (PsaB-Trp(673)) did not produce this effect. The PsaA-W693F mutation also produced spectral changes in the time-resolved EPR spectrum of the P(700)(+) A(1)(-) radical pair, whereas the analogous mutation in PsaB had no observable effect. These observations indicate that the A(1)(-) phylloquinone radical observed by EPR occupies the phylloquinone binding site containing PsaA-Trp(693). However, mutation of either tryptophan accelerated charge recombination from the terminal Fe-S clusters. PMID- 11489880 TI - Caffeine sensitizes human H358 cell line to p53-mediated apoptosis by inducing mitochondrial translocation and conformational change of BAX protein. AB - The mechanisms involved in p53-mediated cell death remain controversial. In the present study, we investigated this cell death pathway by stably transfecting the p53-null H358 cell line with a tetracycline-dependent wild type p53-expressing vector. Restoration of p53 triggered a G(2)/M cell cycle arrest and enhanced BAX protein expression, without inducing apoptosis or potentiating the cytotoxic effect of etoposide, vincristine, and cis-platinum. Accordingly, overexpression of BAX in H358 cells, through stable transfection of a tetracycline-regulated expression vector, did not induce cell death. Interestingly, the methylxanthine caffeine (4 mm) promoted the translocation of BAX from the cytosol to the mitochondria. In the setting of an overexpression of BAX, caffeine induced a conformational change of the protein and apoptosis. The consequences of caffeine were independent of its cell cycle-related activities. All together, caffeine synergizes with p53 for inducing cell death through a cell cycle-independent mechanism, involving mitochondrial translocation and conformational change of BAX protein. PMID- 11489881 TI - Rho and Rho-associated kinase modulate the tyrosine kinase PYK2 in T-cells through regulation of the activity of the integrin LFA-1. AB - We have examined the role of the small GTPase Rho and its downstream effector, the Rho-associated kinase (ROCK), in the control of the adhesive and signaling function of the lymphocyte function-associated antigen-1 (LFA-1) integrin in human T-lymphocytes. Inhibition of Rho (either by treatment with C3-exoenzyme or transfection with a dominant-negative form of Rho (N19Rho)) or ROCK (by treatment with Y-27632) results in the following: (a) partial disorganization and aggregation of cortical filamentous actin (F-actin); (b) induction of LFA-1 mediated cellular adhesion to the LFA-1 ligand intercellular adhesion molecule-1 (ICAM-1) through a mechanism involving clustering of LFA-1 molecules, rather than alterations in the level of expression or in the affinity state of this integrin; and (c) induction of cellular polarization and activation of the tyrosine kinase PYK2. Transfection of T-cells with a constitutively active form of Rho (V14Rho) blocks the clustering of LFA-1 on the membrane and the LFA-1-mediated activation of PYK2. Importantly, the activation of PYK2 caused by inhibition of Rho or ROCK takes place only when the T-cells are plated onto ICAM-1 but not when they are either prevented from interacting with ICAM-1 with anti-LFA-1 blocking antibodies or when they are plated on the nonspecific poly-l-lysine substrate. These results indicate that the small GTPase Rho regulates the tyrosine kinase PYK2 in T-cells through the F-actin-mediated control of the activity of the integrin LFA-1. These findings represent a novel paradigm for the regulation of the activity of a cytoplasmic tyrosine kinase by the small GTPase Rho. PMID- 11489882 TI - Rac1 inhibits myogenic differentiation by preventing the complete withdrawal of myoblasts from the cell cycle. AB - The small GTPase protein Rac1 is involved in a wide range of biological processes, yet its role in cell differentiation is mostly unknown. Here we show that Rac1 activity is high in proliferating myoblasts and decreases during the differentiation process. To analyze the involvement of Rac1 in muscle differentiation, different forms of the protein were expressed in muscle cells. A constitutively activated form of Rac1 (Rac1Q61L) inhibited the activity of MyoD in promoting muscle differentiation, whereas a dominant negative form of Rac1 (Rac1T17N) induced the activity of MyoD in promoting muscle differentiation. Expression of Rac1T17N imposed myogenic differentiation on myoblasts growing under mitogenic conditions. In inquiring whether Rac1 affected the withdrawal of myoblasts from the cell cycle, we analyzed the expression of cyclin D1 and p21(WAF1) and the phosphorylation state of the retinoblastoma protein. According to these markers and bromodeoxyuridine incorporation, C2 myoblasts expressing Rac1T17N exited the cell cycle earlier than control C2 cells. Myoblasts expressing Rac1Q61L did not permanently withdraw from the cell cycle. An indication of the possible involvement of the mitogen-activated protein kinase (MAPK) pathway in Rac1-mediated myoblast proliferation was obtained by the use of MAPK kinase inhibitors U0126 and PD098059. These inhibitors arrested C2-Rac1Q61L cell cycling. Taken together, our results show that Rac1 activation interferes with myoblast exit from the cell cycle via or in concert with the MAPK pathway. PMID- 11489883 TI - The mechanism of superoxide scavenging by Archaeoglobus fulgidus neelaredoxin. AB - Neelaredoxin is a mononuclear iron protein widespread among prokaryotic anaerobes and facultative aerobes, including human pathogens. It has superoxide scavenging activity, but the exact mechanism by which this process occurs has been controversial. In this report, we present the study of the reaction of superoxide with the reduced form of neelaredoxin from the hyperthermophilic archaeon Archaeoglobus fulgidus by pulse radiolysis. This protein reduces superoxide very efficiently (k = 1.5 x 10(9) m(-1)s(-1)), and the dismutation activity is rate limited, in steady-state conditions, by the much slower superoxide oxidation step. These data show unambiguously that the superfamily of neelaredoxin-like proteins (including desulfoferrodoxin) presents a novel type of reactivity toward superoxide, a result of particular relevance for the understanding of both oxygen stress response mechanisms and, in particular, how pathogens may respond to the oxidative burst produced by the defense cells in eukaryotes. The actual in vivo functioning of these enzymes will depend strongly on the cell redox status. Further insight on the catalytic mechanism was obtained by the detection of a transient intermediate ferric species upon oxidation of neelaredoxin by superoxide, detectable by visible spectroscopy with an absorption maximum at 610 nm, blue-shifted approximately 50 nm from the absorption of the resting ferric state. The role of the iron sixth ligand, glutamate-12, in the reactivity of neelaredoxin toward superoxide was assessed by studying two site-directed mutants: E12Q and E12V. PMID- 11489884 TI - Characterization of CD36/LIMPII homologues in Dictyostelium discoideum. AB - The CD36/LIMPII family is ubiquitously expressed in higher eukaryotes and consists of integral membrane proteins that have in part been characterized as cell adhesion receptors, scavenger receptors, or fatty acid transporters. However, no physiological role has been defined so far for the members of this family that localize specifically to vesicular compartments rather than to the cell surface, namely lysosomal integral membrane protein type II (LIMPII) from mammals and LmpA from the amoeba Dictyostelium discoideum. LmpA, the first described CD36/LIMPII homologue from lower eukaryotes, has initially been identified as a suppressor of the profilin-minus phenotype. We report the discovery and initial characterization of two new CD36/LIMPII-related proteins, both of which share several features with LmpA: (i) their size is considerably larger than that of the CD36/LIMPII proteins from higher eukaryotes; (ii) they show the characteristic "hairpin" topology of this protein family; (iii) they are heavily N-glycosylated; and (iv) they localize to vesicular structures of putative endolysosomal origin. However, they show intriguing differences in their developmental regulation and exhibit different sorting signals of the di-leucine or tyrosine-type in their carboxyl-terminal tail domains. These features make them promising candidates as a paradigm for the study of the function and evolution of the as yet poorly understood CD36/LIMPII proteins. PMID- 11489885 TI - Thyroid hormone-deficient period prior to the onset of hearing is associated with reduced levels of beta-tectorin protein in the tectorial membrane: implication for hearing loss. AB - The genes for alpha- and beta-tectorin encode the major non-collagenous proteins of the tectorial membrane. Recently, a targeted deletion of the mouse alpha tectorin gene was found to cause loss of cochlear sensitivity (). Here we describe that mRNA levels for beta-tectorin, but not alpha-tectorin, are significantly reduced in the cochlear epithelium under constant hypothyroid conditions and that levels of beta-tectorin protein in the tectorial membrane are lower. A delay in the onset of thyroid hormone supply prior to onset of hearing, recently described to result in permanent hearing defects and loss of active cochlear mechanics (), can also lead to permanently reduced beta-tectorin protein levels in the tectorial membrane. beta-Tectorin protein levels remain low in the tectorial membrane up to one year after the onset of thyroid hormone supply has been delayed until postnatal day 8 or later and are associated with an abnormally structured tectorial membrane and the loss of active cochlear function. These data indicate that a simple delay in thyroid hormone supply during a critical period of development can lead to low beta-tectorin levels in the tectorial membrane and suggest for the first time that beta-tectorin may be required for development of normal hearing. PMID- 11489886 TI - 4'-phosphopantetheine transfer in primary and secondary metabolism of Bacillus subtilis. AB - 4'-Phosphopantetheine transferases (PPTases) transfer the 4'-phosphopantetheine moiety of coenzyme A onto a conserved serine residue of acyl carrier proteins (ACPs) of fatty acid and polyketide synthases as well as peptidyl carrier proteins (PCPs) of nonribosomal peptide synthetases. This posttranslational modification converts ACPs and PCPs from their inactive apo into the active holo form. We have investigated the 4'-phosphopantetheinylation reaction in Bacillus subtilis, an organism containing in total 43 ACPs and PCPs but only two PPTases, the acyl carrier protein synthase AcpS of primary metabolism and Sfp, a PPTase of secondary metabolism associated with the nonribosomal peptide synthetase for the peptide antibiotic surfactin. We identified and cloned ydcB encoding AcpS from B. subtilis, which complemented an Escherichia coli acps disruption mutant. B. subtilis AcpS and its substrate ACP were biochemically characterized. AcpS also modified the d-alanyl carrier protein but failed to recognize PCP and an acyl carrier protein of secondary metabolism discovered in this study, designated AcpK, that was not identified by the Bacillus genome project. On the other hand, Sfp was able to modify in vitro all acyl carrier proteins tested. We thereby extend the reported broad specificity of this enzyme to the homologous ACP. This in vitro cross-interaction between primary and secondary metabolism was confirmed under physiological in vivo conditions by the construction of a ydcB deletion in a B. subtilis sfp(+) strain. The genes coding for Sfp and its homolog Gsp from Bacillus brevis could also complement the E. coli acps disruption. These results call into question the essential role of AcpS in strains that contain a Sfp-like PPTase and consequently the suitability of AcpS as a microbial target in such strains. PMID- 11489887 TI - Characterization of a novel mammalian SUMO-1/Smt3-specific isopeptidase, a homologue of rat axam, which is an axin-binding protein promoting beta-catenin degradation. AB - A novel SUMO-1/Smt3-specific isopeptidase, SMT3IP2/Axam2 (Smt3-specific isopeptidase 2), was cloned and characterized. The catalytic domains in the carboxyl-terminal region were very much similar to those of other SUMO-1/Smt3 specific proteases, but the amino-terminal part was quite different. The enzyme specifically bound to Smt3a and Smt3b but not to SUMO-1. The SMT3IP2 expressed by Escherichia coli could cleave SUMO-1, Smt3a, or Smt3b from a SUMO-1/RanGAP1, Smt3a/RanGAP1, or Smt3b/RanGAP1 conjugate, respectively, and had the activity of a carboxyl-terminal hydrolase to produce a glycine residue in the carboxyl terminus of these ubiquitin-like proteins. The sequence data indicated that the amino acid sequence of SMT3IP2 was mostly identical to that of rat Axam, which binds to Axin and promotes the degradation of beta-catenin, although its amino terminal region was much shorter than that of Axam. Therefore, we designated this isopeptidase SMT3IP2/Axam2. When human SW480 cells were transfected with wild type SMT3IP2/Axam2, the beta-catenin disappeared. When the cells were transfected with the SMT3IP2/Axam2 C500A mutant, which had neither isopeptidase nor carboxyl terminal hydrolase activity, or with the 1-352 mutant, which lacked the catalytic domain of the enzyme, again the beta-catenin disappeared, indicating that the enzyme activities were not necessary for the instability of beta-catenin in this transfection assay system and that its competition with Dvl for binding to Axin may be important for the instability of beta-catenin as suggested previously for Axam (Kadoya, T., Kishida, S., Fukui, A., Hinoi, T., Michiue, T., Asashima, M., and Kikuchi, A. (2000) J. Biol. Chem. 275, 37030-37037). The involvement of its enzyme activities in the Wnt signaling pathway remains to be elucidated. PMID- 11489889 TI - Molecular cloning and expression of human bile acid beta-glucosidase. AB - A novel microsomal beta-glucosidase was recently purified and characterized from human liver that catalyzes the hydrolysis of bile acid 3-O-glucosides as endogenous compounds. The primary structure of this bile acid beta-glucosidase was deduced by cDNA cloning on the basis of the amino acid sequences of peptides obtained from the purified enzyme by proteinase digestion. The isolated cDNA comprises 3639 base pairs containing 524 nucleotides of 5'-untranslated and 334 nucleotides of 3'-untranslated sequences including the poly(A) tail. The open reading frame predicts a 927-amino acid protein with a calculated M(r) of 104,648 containing one putative transmembrane domain. Data base searches revealed no homology with any known glycosyl hydrolase or other functionally identified protein. The cDNA sequence was found with significant identity in the human chromosome 9 clone RP11-112J3 of the human genome project. The recombinant enzyme was expressed in a tagged form in COS-7 cells where it displayed bile acid beta glucosidase activity. Northern blot analysis of various human tissues revealed high levels of expression of the bile acid beta-glucosidase mRNA (3.6-kilobase message) in brain, heart, skeletal muscle, kidney, and placenta and lower levels of expression in the liver and other organs. PMID- 11489888 TI - ActA from Listeria monocytogenes can interact with up to four Ena/VASP homology 1 domains simultaneously. AB - The facultative intracellular human pathogenic bacterium Listeria monocytogenes actively recruits host actin to its surface to achieve motility within infected cells. The bacterial surface protein ActA is solely responsible for this process by mimicking fundamental steps of host cell actin dynamics. ActA, a modular protein, contains an N-terminal actin nucleation site and a central proline-rich motif of the 4-fold repeated consensus sequence FPPPP (FP(4)). This motif is specifically recognized by members of the Ena/VASP protein family. These proteins additionally recruit the profilin-G-actin complex increasing the local concentration of G-actin close to the bacterial surface. By using analytical ultracentrifugation, we show that a single ActA molecule can simultaneously interact with four Ena/VASP homology 1 (EVH1) domains. The four FP(4) sites have roughly equivalent affinities with dissociation constants of about 4 microm. Mutational analysis of the FP(4) motifs indicate that the phenylalanine is mandatory for ActA-EVH1 interaction, whereas in each case exchange of the third proline was tolerated. Finally, by using sedimentation equilibrium centrifugation techniques, we demonstrate that ActA is a monomeric protein. By combining these results, we formulate a stoichiometric model to describe how ActA enables Listeria to utilize efficiently resources of the host cell microfilament for its own intracellular motility. PMID- 11489890 TI - A coding region determinant of instability regulates levels of manganese superoxide dismutase mRNA. AB - The mitochondria-localized manganese superoxide dismutase (MnSOD), serves a key cytoprotective role against reactive oxygen species arising from a variety of cellular processes and immunological stresses. Previous data from our laboratory suggest that the regulation of the rat MnSOD gene may occur not only at the transcriptional but quite possibly at the post-transcriptional level. To verify this hypothesis, we have attempted to identify regions within the rat MnSOD cDNA that may be functionally involved in regulating the stability of the mRNA. Using a c-fos-based promoter activation system, we have identified an approximately 280 nucleotide fragment within the MnSOD mRNA coding region that, when fused to a rabbit beta-globin gene, destabilizes the normally stable beta-globin mRNA. This cis-directed destabilization phenomenon confers its effects independent of position and stimulus. Most importantly, the MnSOD coding region determinant functions when placed in the 3'-untranslated region of the beta-globin transcript, demonstrating its activity in the absence of ribosome transit. We feel that these data provide a mechanistic basis for both the basal and stimulus dependent post-transcriptional regulation of MnSOD. PMID- 11489891 TI - MKP-7, a novel mitogen-activated protein kinase phosphatase, functions as a shuttle protein. AB - Mitogen-activated protein kinase (MAPK) phosphatases (MKPs) negatively regulate MAPK activity. In the present study, we have identified a novel MKP, designated MKP-7, and mapped it to human chromosome 12p12. MKP-7 possesses a long C-terminal stretch containing both a nuclear export signal and a nuclear localization signal, in addition to the rhodanese-like domain and the dual specificity phosphatase catalytic domain, both of which are conserved among MKP family members. When expressed in mammalian cells MKP-7 protein was localized exclusively in the cytoplasm, but this localization became exclusively nuclear following leptomycin B treatment or introduction of a mutation in the nuclear export signal. These findings indicate that MKP-7 is the first identified leptomycin B-sensitive shuttle MKP. Forced expression of MKP-7 suppressed activation of MAPKs in COS-7 cells in the order of selectivity, JNK p38 > ERK. Furthermore, a mutant form MKP-7 functioned as a dominant negative particularly against the dephosphorylation of JNK, suggesting that MKP-7 works as a JNK specific phosphatase in vivo. Co-immunoprecipitation experiments and histological analysis suggested that MKP-7 determines the localization of MAPKs in the cytoplasm. PMID- 11489892 TI - Unreliability of the cytochrome c-enhanced green fluorescent fusion protein as a marker of cytochrome c release in cells that overexpress Bcl-2. AB - A cytochrome c-enhanced green fluorescent protein chimera (cyt-c.EGFP) was used to monitor the release of cytochrome c from mitochondria in Bcl-2-negative and Bcl-2-positive MDA-MB-468 breast cancer cells. A comparison was made with the intracellular distribution of endogenous cytochrome c based on Western blotting of cell fractions and immunocytochemistry. The release of endogenous cytochrome c from mitochondria into the cytoplasm was detected in Bcl-2-negative cells treated with the kinase inhibitor staurosporine or the calcium-ATPase inhibitor thapsigargin. No release of endogenous cytochrome c was evident in Bcl-2-positive cells, consistent with earlier evidence that Bcl-2 overexpression inhibits cytochrome c release from mitochondria. Cyt-c.EGFP appeared to be localized to the mitochondria in Bcl-2-negative cells and to be released into the cytoplasm following treatment with either staurosporine or thapsigargin. However, in Bcl-2 positive cells the pattern of distribution of cytochrome c-EGFP was inconsistent with that of endogenous cytochrome c, due to accumulation of both cyt-c.EGFP and free EGFP in the cytoplasm of both treated and untreated cells. In summary, cyt c.EGFP may be useful for monitoring cytochrome c release in living cells that do not express high levels of Bcl-2 but is an unreliable marker of cytochrome c release in cells that overexpress Bcl-2. PMID- 11489893 TI - Functional analysis of the hydrophobic patch on nuclear transport factor 2 involved in interactions with the nuclear pore in vivo. AB - Nuclear transport factor 2 (NTF2) is a small homodimeric protein that interacts simultaneously with both RanGDP and FxFG nucleoporins. The interaction between NTF2 and Ran is essential for the import of Ran into the nucleus. Here we use mutational analysis to dissect the in vivo role of the interaction between NTF2 and nucleoporins. We identify a series of surface residues that form a hydrophobic patch on NTF2, which when mutated disrupt the NTF2-nucleoporin interaction. Analysis of these mutants in vivo demonstrates that the strength of this interaction can be significantly reduced without affecting cell viability. However, cells cease to be viable if the interaction between NTF2 and nucleoporins is abolished completely, indicating that this interaction is essential for the function of NTF2 in vivo. In addition, we have isolated a dominant negative mutant of NTF2, N77Y, which has increased affinity for nucleoporins. Overexpression of the N77Y protein blocks nuclear protein import and concentrates Ran at the nuclear rim. These data support a mechanism in which NTF2 interacts transiently with FxFG nucleoporins to translocate through the pore and import RanGDP into the nucleus. PMID- 11489894 TI - The lipid/protein interface as xenobiotic target site: kinetic analysis of the nicotinic acetylcholine receptor. AB - Membrane proteins are known to be solvated and functionally activated by a fixed number of lipid molecules whose multiple binding can be described by Adair-type binding equations. Lipophilic xenobiotics such as general anesthetics may act by competitive displacement of protein-bound lipids. A kinetic equation is now presented for various binding stoichiometries of lipid and xenobiotic, and microscopic binding constants of anesthetics and organic solvents are derived from two independent assay systems for the enhancement of agonist binding to the nicotinic acetylcholine receptor. These constants lead to the first available free energy estimate (-6.4 kcal/mol) for the binding of membrane lipid to an integral membrane protein. PMID- 11489895 TI - Molecular characterization of the mouse Tem1/endosialin gene regulated by cell density in vitro and expressed in normal tissues in vivo. AB - Human tumor endothelial marker 1/endosialin (TEM1/endosialin) was recently identified as a novel tumor endothelial cell surface marker potentially involved in angiogenesis, although no specific function for this novel gene has been assigned so far. It was reported to be expressed in tumor endothelium but not in normal endothelium with the exception of perhaps the corpus luteum. Here we describe the cDNA and genomic sequences for the mouse Tem1/endosialin homolog, the identification and characterization of its promoter region, and an extensive characterization of its expression pattern in murine and human tissues and murine cell lines in vitro. The single copy gene that was mapped to chromosome 19 is intronless and encodes a 92-kDa protein that has 77.5% overall homology to the human protein. The remarkable findings are 1) this gene is ubiquitously expressed in normal human and mouse somatic tissues and during development, and 2) its expression at the mRNA level is density-dependent and up-regulated in serum starved cells. In vitro, its expression is limited to cells of embryonic, endothelial, and preadipocyte origin, suggesting that the wide distribution of its expression in vivo is due to the presence of vascular endothelial cells in all the tissues. The ubiquitous expression in vivo is in contrast to previously reported expression limited to corpus luteum and highly angiogenic tissues such as tumors and wound tissue. PMID- 11489896 TI - Role of the deafness dystonia peptide 1 (DDP1) in import of human Tim23 into the inner membrane of mitochondria. AB - Tim8 and Tim13 of yeast belong to a family of evolutionary conserved zinc finger proteins that are organized in hetero-oligomeric complexes in the mitochondrial intermembrane space. Mutations in DDP1 (deafness dystonia peptide 1), the human homolog of Tim8, are associated with the Mohr-Tranebjaerg syndrome, a progressive neurodegenerative disorder. We show that DDP1 acts with human Tim13 in a complex in the intermembrane space. The DDP1.hTim13 complex is in direct contact with translocation intermediates of human Tim23 in mammalian mitochondria. The human DDP1.hTim13 complex complements the function of the TIM8.13 complex in yeast and facilitates import of yeast and human Tim23. Thus, the pathomechanism underlying the Mohr-Tranebjaerg syndrome may involve an impaired biogenesis of the human TIM23 complex causing severe pleiotropic mitochondrial dysfunction. PMID- 11489897 TI - The smooth muscle myosin heavy chain gene exhibits smooth muscle subtype selective modular regulation in vivo. AB - Previous studies in our laboratory demonstrated that the transgene consisting of the -4.2 to +11.6 kilobase (kb) region of the smooth muscle (SM) myosin heavy chain (MHC) gene was expressed in virtually all SM tissue types in vivo in transgenic mice and that the multiple CArG elements within this region were differentially required in SMC subtypes, implying that the SM-MHC gene was controlled by multiple transcriptional regulatory modules. To investigate this hypothesis, we analyzed specific regulatory regions within the SM-MHC -4.2 to +11.6 kb region by a combination of deletion analyses of various SM-MHC transgenes as well as by DNaseI hypersensitivity assays and in vivo footprinting in intact SMC tissues. The results showed that SM-MHC transgene expression depended on a large number of required regulatory modules that were widely spread over the -4.2 to +11.6 region. Moreover, the results revealed several unexpected novel features of regulation of the SM-MHC gene including: 1) unique combinations of regulatory modules were required for SM-MHC expression in different SMC subtypes; 2) repressor modules as well as activator modules were both critical for SMC specificity of the gene; 3) certain modules were required in certain contexts but were dispensable in others within a given SMC-subtype (i.e. the net activity of the module was determined by interaction between modules not simply by the sum of module activities); and 4) we identified a highly conserved 200 base pair transcriptional regulatory module at +8 kb that was required in the large arteries but dispensable in the coronary arteries and airways in transgenic mice and contained multiple potential cis-elements that were occupied by nuclear proteins in the intact aorta based on in vivo footprinting. Taken together, the results suggest a model of complex modular control of expression of the SM-MHC gene that varies between SMC subtypes. Moreover, the studies establish the possibility of designing derivatives of the SM-MHC promoter that might be used for targeting gene expression to specific SMC subtypes in vivo. PMID- 11489898 TI - Parathyroid hormone-related protein promotes quiescence and survival of serum deprived chondrocytes by inhibiting rRNA synthesis. AB - Parathyroid hormone-related protein (PTHrP) was initially recognized for its ability to promote parathyroid hormone-like bioactivity in kidney, bone, and squamous epithelial cells. PTHrP is a multifunctional protein in which bioactivity is mediated by two distinct pathways. Its classic parathyroid hormone like activity results from binding of its amino terminus to cell surface PTH1R and activation of signal transduction pathways. Another less well recognized pathway involves translocation of PTHrP to the nucleus via a mid-region bipartite nuclear targeting sequence (NTS), similar in structure and function to those found in retroviral regulatory proteins. PTHrP was identified in the nucleus of several different cell types in vivo and in vitro, where it has been implicated in cell cycle progression, cellular differentiation, and apoptosis. In previous work we showed that nuclear translocation of PTHrP enhanced the survival of serum deprived chondrogenic cells, associated with RNA, and localized to a region of the nucleus rich in complexes of newly transcribed ribosomal RNA and protein. In this work we have used two chondrogenic cell lines, CFK2 (PTH1R+) and 27m21 (PTH1R-) to further explore mechanisms whereby PTHrP rescues immature chondrocytes from apoptosis. Endogenous PTHrP and exogenous PTHrP NTS peptide protected serum-deprived cells from apoptosis, in the presence and absence of PTH1R. The survival of cells expressing PTHrP and those treated with PTHrP NTS peptide was associated with a rapid shift into G(o)/G1 accompanied by a significant down-regulation of rRNA synthesis and a decrease in the number of actively translating polyribosome complexes. Together with our previous observations, this work predicts a role for PTHrP in modulating ribosome biogenesis and preventing chondrogenic cells from progressing through the cell cycle in an unfavorable environment. PMID- 11489899 TI - The 5'-untranslated region of the FMR1 message facilitates translation by internal ribosome entry. AB - Fragile X syndrome, the leading heritable form of mental impairment, is generally caused by large expansions of a CGG repeat in the promoter region of the FMR1 gene followed by transcriptional silencing. However, there is growing evidence that translation of the FMR1 message is also impaired, presumably because of the expanded CGG element in the 5'-untranslated region (5'-UTR) of the FMR1 message. To study the properties of the FMR1 5'-UTR, deletions were generated within a normal 5'-UTR with 16 CGG repeats for both monocistronic and dicistronic (luciferase) reporter constructs. Transient transfection experiments revealed a approximately 20-nucleotide region upstream of the CGG repeat element that functions as an internal ribosome entry site (IRES). The normal CGG element itself does not appear to influence the efficiency of IRES-mediated stimulation of downstream reporter activity (approximately 18-fold over controls). Additional controls indicate that the enhanced activity of the downstream reporter is not due to readthrough from the upstream cistron, nor is it due to translation of cryptic monocistronic transcripts. The role of the FMR1 IRES element is not known at present; however, by analogy to other IRES-containing mRNAs expressed in neurons, the FMR1 IRES element may help to promote translation in dendrites. PMID- 11489900 TI - Heterodimerization of calcium sensing receptors with metabotropic glutamate receptors in neurons. AB - Calcium sensing (CaR) and Group I metabotropic glutamate receptors exhibit overlapping expression patterns in brain, and share common signal transduction pathways. To determine whether CaR and Group I metabotropic glutamate receptors (mGluRs) (mGluR1alpha and mGluR5) can form heterodimers, we immunoprecipitated CaR from bovine brain and observed co-precipitation of mGluR1alpha. CaR and mGluR1alpha co-localize in hippocampal and cerebellar neurons, but are expressed separately in other brain regions. In vitro transfection studies in HEK-293 cells established the specificity and disulfide-linked nature of the CaR:mGluR1alpha (CaR:mGluR5) interactions. CaR:mGluR1alpha (CaR:mGluR5) heterodimers exhibit altered trafficking via Homer 1c when compared with CaR:CaR homodimers. CaR becomes sensitive to glutamate-mediated internalization when present in CaR:mGluR1alpha heterodimers. These results demonstrate cross-family covalent heterodimerization of CaR with Group I mGluRs, and increase the potential role(s) for CaR in modulating neuronal function. PMID- 11489901 TI - Three-dimensional structure of a hyperthermophilic 5'-deoxy-5' methylthioadenosine phosphorylase from Sulfolobus solfataricus. AB - The structure of 5'-deoxy-5'-methylthioadenosine phosphorylase from Sulfolobus solfataricus (SsMTAP) has been determined alone, as ternary complexes with sulfate plus substrates 5'-deoxy-5'-methylthioadenosine, adenosine, or guanosine, or with the noncleavable substrate analog Formycin B and as binary complexes with phosphate or sulfate alone. The structure of unliganded SsMTAP was refined at 2.5 A resolution and the structures of the complexes were refined at resolutions ranging from 1.6 to 2.0 A. SsMTAP is unusual both for its broad substrate specificity and for its extreme thermal stability. The hexameric structure of SsMTAP is similar to that of purine-nucleoside phosphorylase (PNP) from Escherichia coli, however, only SsMTAP accepts 5'-deoxy-5'-methylthioadenosine as a substrate. The active site of SsMTAP is similar to that of E. coli PNP with 13 of 18 nearest residues being identical. The main differences are at Thr(89), which corresponds to serine in E. coli PNP, and Glu(163), which corresponds to proline in E. coli PNP. In addition, a water molecule is found near the purine N 7 position in the guanosine complex of SsMTAP. Thr(89) is near the 5'-position of the nucleoside and may account for the ability of SsMTAP to accept either hydrophobic or hydrophilic substituents in that position. Unlike E. coli PNP, the structures of SsMTAP reveal a substrate-induced conformational change involving Glu(163). This residue is located at the interface between subunits and swings in toward the active site upon nucleoside binding. The high-resolution structures of SsMTAP suggest that the transition state is stabilized in different ways for 6 amino versus 6-oxo substrates. SsMTAP has optimal activity at 120 degrees C and retains full activity after 2 h at 100 degrees C. Examination of the three dimensional structure of SsMTAP suggests that unlike most thermophilic enzymes, disulfide linkages play a key in role in its thermal stability. PMID- 11489902 TI - Gleevec (STI571) influences metabolic enzyme activities and glucose carbon flow toward nucleic acid and fatty acid synthesis in myeloid tumor cells. AB - Chronic myeloid leukemia cells contain a constitutively active Bcr-Abl tyrosine kinase, the target protein of Gleevec (STI571) phenylaminopyrimidine class protein kinase inhibitor. Here we provide evidence for metabolic phenotypic changes in cultured K562 human myeloid blast cells after treatment with increasing doses of STI571 using [1,2-13C2]glucose as the single tracer and biological mass spectrometry. In response to 0.68 and 6.8 microm STI571, proliferation of Bcr-Abl-positive K562 cells showed a 57% and 74% decrease, respectively, whereas glucose label incorporation into RNA decreased by 13.4% and 30.1%, respectively, through direct glucose oxidation, as indicated by the decrease in the m1/Sigma(m)n ratio in RNA. Based on the in vitro proliferation data, the IC50 of STI571 in K562 cultures is 0.56 microm. The decrease in 13C label incorporation into RNA ribose was accompanied by a significant fall in hexokinase and glucose-6-phosphate 1-dehydrogenase activities. The activity of transketolase, the enzyme responsible for nonoxidative ribose synthesis in the pentose cycle, was less affected, and there was a relative increase in glucose carbon incorporation into RNA through nonoxidative synthesis as indicated by the increase in the m2/Sigma(m)n ratio in RNA. The restricted use of glucose carbons for de novo nucleic acid and fatty acid synthesis by altering metabolic enzyme activities and pathway carbon flux of the pentose cycle constitutes the underlying mechanism by which STI571 inhibits leukemia cell glucose substrate utilization and growth. The administration of specific hexokinase/glucose-6 phosphate 1-dehydrogenase inhibitor anti-metabolite substrates or competitive enzyme inhibitor compounds, alone or in combination, should be explored for the treatment of STI571-resistant advanced leukemias as well as that of Bcr-Abl negative human malignancies. PMID- 11489903 TI - Polyamine regulation of ribosome pausing at the upstream open reading frame of S adenosylmethionine decarboxylase. AB - Synthesis of S-adenosylmethionine decarboxylase (AdoMetDC), a key regulated enzyme in the pathway of polyamine biosynthesis, is feedback-controlled at the level of translation by spermidine and spermine. The peptide product of an upstream open reading frame (uORF) in the mRNA is solely responsible for polyamine regulation of AdoMetDC translation. Using a primer extension inhibition assay and in vitro protein synthesis reactions, we found ribosomes paused at or close to the termination codon of the uORF. This pause was greatly diminished with the altered uORFs' sequences that abolish uORF regulation in vivo. The half life of the ribosome pause was related to the concentration of polyamines present but was unaffected by magnesium concentration. Furthermore, inhibition of translation initiation at a reporter gene placed downstream of the AdoMetDC uORF directly correlated with the stability of the ribosome pause at the uORF. These observations are consistent with a model in which regulation of ribosome pausing at the uORF by polyamines controls ribosome access to the downstream AdoMetDC reading frame. PMID- 11489904 TI - A membrane protein enriched in endoplasmic reticulum exit sites interacts with COPII. AB - Although all mammalian COPII components have now been cloned, little is known of their interactions with other regulatory proteins involved in exit from the endoplasmic reticulum (ER). We report here that a mammalian protein (Yip1A) that is about 31% identical to S. cerevisiae and which interacts with and modulates COPII-mediated ER-Golgi transport. Yip1A transcripts are ubiquitously expressed. Transcripts of a related mammalian homologue, Yip1B, are found specifically in the heart. Indirect immunofluorescence microscopy revealed that Yip1A is localized to vesicular structures that are concentrated at the perinuclear region. The structures marked by Yip1A co-localized with Sec31A and Sec13, components of the COPII coat protein complex. Immunoelectron microscopy also showed that Yip1A co-localizes with Sec13 at ER exit sites. Overexpression of the hydrophilic N terminus of Yip1A arrests ER-Golgi transport of the vesicular stomatitis G protein and causes fragmentation and dispersion of the Golgi apparatus. A glutathione S-transferase fusion protein with the hydrophilic N terminus of Yip1A (GST-Yip1A) is able to bind to and deplete vital components from rat liver cytosol that is essential for in vitro vesicular stomatitis G transport. Peptide sequence analysis of cytosolic proteins that are specifically bound to GST-Yip1A revealed, among other proteins, mammalian COPII components Sec23 and Sec24. A highly conserved domain at the N terminus of Yip1A is required for Sec23/Sec24 interaction. Our results suggest that Yip1A is involved in the regulation of ER-Golgi traffic at the level of ER exit sites. PMID- 11489905 TI - Polyunsaturated eicosapentaenoic acid displaces proteins from membrane rafts by altering raft lipid composition. AB - Polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (20:5 (n-3)) inhibit T lymphocyte activation probably by displacing acylated signaling proteins from membrane lipid rafts. Under physiological conditions, saturated fatty acyl residues of such proteins partition into the cytoplasmic membrane lipid leaflet with high affinity for rafts that are enriched in saturated fatty acyl-containing lipids. However, the biochemical alteration causing displacement of acylated proteins from rafts in PUFA-treated T cells is still under debate but could principally be attributed to altered protein acylation or changes in raft lipid composition. We show that treatment of Jurkat T cells with polyunsaturated eicosapentaenoic acid (20:5 (n-3)) results in marked enrichment of PUFAs (20:5; 22:5) in lipids from isolated rafts. Moreover, PUFAs were significantly incorporated into phosphatidylethanolamine that predominantly resides in the cytoplasmic membrane lipid leaflet. Notably, palmitate-labeled Src family kinase Lck and the linker for activation of T cells (LAT) were both displaced from lipid rafts indicating that acylation by PUFAs is not required for protein displacement from rafts in PUFA-treated T cells. In conclusion, these data provide strong evidence that displacement of acylated proteins from rafts in PUFA-treated T cells is predominantly due to altered raft lipid composition. PMID- 11489906 TI - Ligand binding characteristics of CXCR4 incorporated into paramagnetic proteoliposomes. AB - The G protein-coupled receptor CXCR4 is a coreceptor, along with CD4, for the human immunodeficiency virus type 1 (HIV-1) and has been implicated in breast cancer metastasis. We studied the binding of the HIV-1 gp120 envelope glycoprotein (gp) to CXCR4 but found that the gp120s from CXCR4-using HIV-1 strains bound nonspecifically to several cell lines lacking human CXCR4 expression. Therefore, we constructed paramagnetic proteoliposomes (CXCR4-PMPLs) containing pure, native CXCR4. CXCR4-PMPLs specifically bound the natural ligand, SDF-1alpha, and the gp120s from CXCR4-using HIV-1 strains. Conformation-dependent anti-CXCR4 antibodies and the CXCR4 antagonist AMD3100 blocked HIV-1 gp120 binding to CXCR4-PMPLs. The gp120-CXCR4 interaction was blocked by anti-gp120 antibodies directed against the third variable (V3) loop and CD4-induced epitopes, structures that have also been implicated in the binding of gp120 to the other HIV-1 coreceptor, CCR5. Compared with the binding of R5 HIV-1 gp120s to CCR5, the gp120-CXCR4 interaction exhibited a lower affinity (K(d) = 200 nm) and was dependent upon prior CD4 binding, even at low temperature. Thus, although similar regions of X4 and R5 HIV-1 gp120s appear to be involved in binding CXCR4 and CCR5, respectively, differences exist in nonspecific binding to cell surfaces, affinity for the chemokine receptor, and CD4 dependence at low temperature. PMID- 11489907 TI - Sak serine-threonine kinase acts as an effector of Tec tyrosine kinase. AB - The murine sak gene encodes a putative serine-threonine kinase which is homologous to the members of the Plk/Polo family. Although Sak protein is presumed to be involved in cell growth mechanism, efforts have failed to demonstrate its kinase activity. Little has been, therefore, elucidated how Sak is regulated and how Sak contributes to cell proliferation. Tec is a cytoplasmic protein-tyrosine kinase (PTK) which becomes activated by the stimulation of cytokine receptors, lymphocyte surface antigens, heterotrimeric G protein-linked receptors, and integrins. To clarify the in vivo function of Tec, we have tried to isolate the second messengers of Tec by using the yeast two-hybrid screening. One of such Tec-binding proteins turned out to be Sak. In human kidney 293 cells, Sak became tyrosine-phosphorylated by Tec, and the serine-threonine kinase activity of Sak was detected only under the presence of Tec, suggesting Sak to be an effector molecule of Tec. In addition, Tec activity efficiently protects Sak from the "PEST" sequence-dependent proteolysis. Internal deletion of the PEST sequences led to the stabilization of Sak proteins, and expression of these mutants acted suppressive to cell growth. Our data collectively supports a novel role of Sak acting in the PTK-mediated signaling pathway. PMID- 11489908 TI - Interaction of Met297 in the seventh transmembrane segment of the tachykinin NK2 receptor with neurokinin A. AB - We report the use of thiol chemistry to define specific and reversible disulfide interactions of Cys-substituted NK2 receptor mutants with analogues of neurokinin A (NKA) containing single cysteine substitutions. The NKA analogues were N biotinylated to facilitate the rapid detection of covalent analogue-receptor interactions utilizing streptavidin reactivity. N-biotinyl-[Tyr1,Cys9]NKA, N biotinyl-[Tyr1,Cys10]NKA were both found to reversibly disulfide bond to the NK2 receptor mutant Met297 --> Cys. This is consistent with the improved affinities of these particular analogues for the Met297 --> Cys receptor as compared with those for the wild-type and Met297 --> Leu receptors. In our three-dimensional model, Met297 occupies the equivalent position in helix 7 to the retinal binding Lys296 in rhodopsin. Binding of the NK2 receptor antagonist [3H]SR 48968 and of 125I-NKA was used to characterize additional receptor mutants. It seems that the aromatic residues Trp99 (helix 3), His198 (helix 5), Tyr266, His267, and Phe270 play an important role in NKA binding as structural determinants. The existence of overlapping SR 48968 and NKA binding sites is also evident. These data suggest that the peptide binding site of the NK2R is at least in part formed by residues buried deep within the transmembrane bundle and that this intramembranous binding domain may correspond to the binding sites for substantially smaller endogenous GPCR ligands. PMID- 11489909 TI - Evidence for selective coupling of alpha 1-adrenergic receptors to phospholipase C-beta 1 in rat neonatal cardiomyocytes. AB - Activation of phospholipase C (PLC) in neonatal rat cardiomyocytes (NCM) generates primarily inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3)) in response to rises in intracellular Ca(2+), or inositol 1,4-bisphosphate (Ins(1,4)P(2)) in response to norepinephrine (NE) (Matkovich, S. J. and Woodcock, E. A. (2000) J. Biol. Chem. 275, 10845-10850). To examine the PLC subtype mediating the alpha(1) adrenergic receptor response, PLC-beta(1) and PLC-beta(3) were overexpressed in NCM using adenoviral infection (Ad-PLC-beta(1) NCM and Ad-PLC-beta(3) NCM, respectively) and PLC responses assessed from [(3)H]inositol phosphate (InsP) generation in the presence of 10 mm LiCl. The [(3)H]InsP response to NE (100 microm) was enhanced in Ad-PLC-beta(1) NCM relative to cells infected with blank virus (Ad-MX NCM), but was reduced in Ad-PLC-beta(3) NCM. In contrast, the [(3)H]InsP response to ATP (100 microm) was not elevated in Ad-PLC-beta(1) NCM, and was enhanced rather than diminished in Ad-PLC-beta(3) NCM, showing that effects of the two PLC-beta isoforms were specific for particular receptor types. PLC-delta(1) overexpression selectively reduced NE-induced [(3)H]InsP responses, without affecting the ATP stimulation. The reduced NE response was associated with a selective loss of PLC-beta(1) expression in Ad-PLC-delta(1) NCM. alpha(1) Adrenergic receptor activation caused phosphorylation of PLC-beta(1) but not PLC beta(3), whereas stimulation by ATP induced phosphorylation of PLC-beta(3) but not PLC-beta(1.) Taken together, these studies provide evidence that NE stimulated InsP generation in NCM is primarily mediated by PLC-beta(1), despite the presence of both PLC-beta(1) and PLC-beta(3) isoforms. PMID- 11489910 TI - Novel differences between two human prion strains revealed by two-dimensional gel electrophoresis. AB - The phenotype of human sporadic prion diseases is affected by patient genotype at codon 129 of the prion protein (PrP) gene, the site of a common methionine/valine polymorphism, and by the type of the scrapie PrP (PrP(Sc)), which likely reflects the prion strain. However, two distinct disease phenotypes, identified as sporadic Creutzfeldt-Jakob disease (M/M2 sCJD) and sporadic fatal insomnia (sFI), share methionine homozygosity at codon 129 and PrP(Sc) type 2. One-dimensional gel electrophoresis and immunoblotting reveal no difference between the M/M2 sCJD and sFI species of PrP(Sc) in gel mobility and glycoform ratio. In contrast, the two-dimensional immunoblot demonstrates that in M/M2 sCJD the full-length PrP(Sc) form is overrepresented and carries glycans that are different from those present in the PrP(Sc) of sFI. Because the altered glycans are detectable only in the PrP(Sc) and not in the normal or cellular PrP (PrP(C)), they are likely to result from preferential conversion to PrP(Sc) of rare PrP(C) glycoforms. This is the first evidence that a qualitative difference in glycans contributes to prion diversity. PMID- 11489911 TI - Neurotrophin p75 receptor is involved in neuronal damage by prion peptide-(106 126). AB - In this work we have investigated the molecular basis of the neuronal damage induced by the prion peptide by searching for a surface receptor whose activation could be the first step of a cascade of events responsible for cell death. By using a human neuroblastoma cell line lacking all the neurotrophin receptors and derived clones expressing the full-length or truncated forms of the low affinity neurotrophin receptor (p75(NTR)), we have been able to demonstrate that the neuronal death induced by the prion protein fragment PrP-(106-126) is an active process mediated by a) the binding of the peptide to the extracellular region of p75(NTR), b) the signaling function of the intracytoplasmic region of the receptor, and c) the activation of caspase-8 and the production of oxidant species. PMID- 11489912 TI - Activated platelets mediate inflammatory signaling by regulated interleukin 1beta synthesis. AB - Platelets release preformed mediators and generate eicosanoids that regulate acute hemostasis and inflammation, but these anucleate cytoplasts are not thought to synthesize proteins or cytokines, or to influence inflammatory responses over time. Interrogation of an arrayed cDNA library demonstrated that quiescent platelets contain many messenger RNAs, one of which codes for interleukin 1beta precursor (pro-IL-1beta). Unexpectedly, the mRNA for IL-1beta and many other transcripts are constitutively present in polysomes, providing a mechanism for rapid synthesis. Platelet activation induces rapid and sustained synthesis of pro IL-1beta protein, a response that is abolished by translational inhibitors. A portion of the IL-1beta is shed in its mature form in membrane microvesicles, and induces adhesiveness of human endothelial cells for neutrophils. Signal-dependent synthesis of an active cytokine over several hours indicates that platelets may have previously unrecognized roles in inflammation and vascular injury. Inhibition of beta3 integrin engagement markedly attenuated the synthesis of IL 1beta, identifying a new link between the coagulation and inflammatory cascades, and suggesting that antithrombotic therapies may also have novel antiinflammatory effects. PMID- 11489913 TI - Junctional adhesion molecule (JAM) binds to PAR-3: a possible mechanism for the recruitment of PAR-3 to tight junctions. AB - At tight junctions (TJs), claudins with four transmembrane domains are incorporated into TJ strands. Junctional adhesion molecule (JAM), which belongs to the immunoglobulin superfamily, is also localized at TJs, but it remains unclear how JAM is integrated into TJs. Immunoreplica electron microscopy revealed that JAM showed an intimate spatial relationship with TJ strands in epithelial cells. In L fibroblasts expressing exogenous JAM, JAM was concentrated at cell-cell adhesion sites, where there were no strand-like structures, but rather characteristic membrane domains free of intramembranous particles were detected. These domains were specifically labeled with anti-JAM polyclonal antibody, suggesting that JAM forms planar aggregates through their lateral self association. Immunofluorescence microscopy and in vitro binding assays revealed that ZO-1 directly binds to the COOH termini of claudins and JAM at its PDZ1 and PDZ3 domains, respectively. Furthermore, another PDZ-containing polarity-related protein, PAR-3, was directly bound to the COOH terminus of JAM, but not to that of claudins. These findings led to a molecular architectural model for TJs: small aggregates of JAM are tethered to claudin-based strands through ZO-1, and these JAM aggregates recruit PAR-3 to TJs. We also discuss the importance of this model from the perspective of the general molecular mechanisms behind the recruitment of PAR proteins to plasma membranes. PMID- 11489914 TI - RNA-mediated interaction of Cajal bodies and U2 snRNA genes. AB - Cajal bodies (CBs) are nuclear structures involved in RNA metabolism that accumulate high concentrations of small nuclear ribonucleoproteins (snRNPs). Notably, CBs preferentially associate with specific genomic loci in interphase human cells, including several snRNA and histone gene clusters. To uncover functional elements involved in the interaction of genes and CBs, we analyzed the expression and subcellular localization of stably transfected artificial arrays of U2 snRNA genes. Although promoter substitution arrays colocalized with CBs, constructs containing intragenic deletions did not. Additional experiments identified factors within CBs that are important for association with the native U2 genes. Inhibition of nuclear export or targeted degradation of U2 snRNPs caused a marked decrease in the levels of U2 snRNA in CBs and strongly disrupted the interaction with U2 genes. Together, the results illustrate a specific requirement for both the snRNA transcripts as well as the presence of snRNPs (or snRNP proteins) within CBs. Our data thus provide significant insight into the mechanism of CB interaction with snRNA loci, strengthening the putative role for this nuclear suborganelle in snRNP biogenesis. PMID- 11489915 TI - Head-to-tail oligomerization of calsequestrin: a novel mechanism for heterogeneous distribution of endoplasmic reticulum luminal proteins. AB - Many proteins retained within the endo/sarcoplasmic reticulum (ER/SR) lumen express the COOH-terminal tetrapeptide KDEL, by which they continuously recycle from the Golgi complex; however, others do not express the KDEL retrieval signal. Among the latter is calsequestrin (CSQ), the major Ca2+-binding protein condensed within both the terminal cisternae of striated muscle SR and the ER vacuolar domains of some neurons and smooth muscles. To reveal the mechanisms of condensation and establish whether it also accounts for ER/SR retention of CSQ, we generated a variety of constructs: chimeras with another similar protein, calreticulin (CRT); mutants truncated of COOH- or NH2-terminal domains; and other mutants deleted or point mutated at strategic sites. By transfection in L6 myoblasts and HeLa cells we show here that CSQ condensation in ER-derived vacuoles requires two amino acid sequences, one at the NH2 terminus, the other near the COOH terminus. Experiments with a green fluorescent protein GFP/CSQ chimera demonstrate that the CSQ-rich vacuoles are long-lived organelles, unaffected by Ca2+ depletion, whose almost complete lack of movement may depend on a direct interaction with the ER. CSQ retention within the ER can be dissociated from condensation, the first identified process by which ER luminal proteins assume a heterogeneous distribution. A model is proposed to explain this new process, that might also be valid for other luminal proteins. PMID- 11489917 TI - Expression of alpha-catenin in alpha-catenin-deficient cells increases resistance to sphingosine-induced apoptosis. AB - Alpha-catenin, an intracellular protein, associates with the COOH-terminal region of cadherin cell adhesion molecules through interactions with either beta-catenin or gamma-catenin (plakoglobin). The full activity of cadherins requires a linkage to the actin cytoskeleton mediated by catenins. We transfected alpha-catenin deficient colon carcinoma cells with a series of alpha-catenin constructs to determine that alpha-catenin expression increases the resistance to apoptosis induced by sphingosine. Two groups of constructs, containing deletions in either the middle segment of the molecule or the COOH terminus, induced morphological changes, cell compaction, and decreases in cell death. In alpha-catenin expressing cells, inhibition of cadherin cell adhesion by treatment with anti-E cadherin antibodies did not decrease the cells viability. alpha-Catenin expression partially suppressed the downregulation of Bcl-xL and the activation of caspase 3. Expression of p27kip1 protein, an inhibitor of cyclin-dependent kinases, was increased by alpha-catenin expression in low density cell cultures. The increased levels of p27kip1 correlated with both increased resistance to cell death and morphological changes in transfectants containing deletion mutants. Transfection-mediated upregulation of p27kip1 decreases sphingosine-induced cell death in alpha-catenin-deficient cells. We postulate that alpha-catenin mediates transduction of signals from the cadherin-catenin complex to regulate the apoptotic cascade via p27kip1. PMID- 11489918 TI - Neuronal survival induced by neurotrophins requires calmodulin. AB - It has been reported that phosphoinositide 3-kinase (PI 3-kinase) and its downstream target, protein kinase B (PKB), play a central role in the signaling of cell survival triggered by neurotrophins (NTs). In this report, we have analyzed the involvement of Ca2+ and calmodulin (CaM) in the activation of the PKB induced by NTs. We have found that reduction of intracellular Ca2+ concentration or functional blockade of CaM abolished NGF-induced activation of PKB in PC12 cells. Similar results were obtained in cultures of chicken spinal cord motoneurons treated with brain-derived neurotrophic factor (BDNF). Moreover, CaM inhibition prevented the cell survival triggered by NGF or BDNF. This effect was counteracted by the transient expression of constitutive active forms of the PKB, indicating that CaM regulates NT-induced cell survival through the activation of the PKB. We have investigated the mechanisms whereby CaM regulates the activation of the PKB, and we have found that CaM was necessary for the proper generation and/or accumulation of the products of the PI 3-kinase in intact cells. PMID- 11489916 TI - A protein interaction map for cell polarity development. AB - Many genes required for cell polarity development in budding yeast have been identified and arranged into a functional hierarchy. Core elements of the hierarchy are widely conserved, underlying cell polarity development in diverse eukaryotes. To enumerate more fully the protein-protein interactions that mediate cell polarity development, and to uncover novel mechanisms that coordinate the numerous events involved, we carried out a large-scale two-hybrid experiment. 68 Gal4 DNA binding domain fusions of yeast proteins associated with the actin cytoskeleton, septins, the secretory apparatus, and Rho-type GTPases were used to screen an array of yeast transformants that express approximately 90% of the predicted Saccharomyces cerevisiae open reading frames as Gal4 activation domain fusions. 191 protein-protein interactions were detected, of which 128 had not been described previously. 44 interactions implicated 20 previously uncharacterized proteins in cell polarity development. Further insights into possible roles of 13 of these proteins were revealed by their multiple two-hybrid interactions and by subcellular localization. Included in the interaction network were associations of Cdc42 and Rho1 pathways with proteins involved in exocytosis, septin organization, actin assembly, microtubule organization, autophagy, cytokinesis, and cell wall synthesis. Other interactions suggested direct connections between Rho1- and Cdc42-regulated pathways; the secretory apparatus and regulators of polarity establishment; actin assembly and the morphogenesis checkpoint; and the exocytic and endocytic machinery. In total, a network of interactions that provide an integrated response of signaling proteins, the cytoskeleton, and organelles to the spatial cues that direct polarity development was revealed. PMID- 11489919 TI - Discovery of a novel murine keratin 6 (K6) isoform explains the absence of hair and nail defects in mice deficient for K6a and K6b. AB - The murine genome is known to have two keratin 6 (K6) genes, mouse K6 (MK6)a and MK6b. These genes display a complex expression pattern with constitutive expression in the epithelia of oral mucosa, hair follicles, and nail beds. We generated mice deficient for both genes through embryonic stem cell technology. The majority of MK6a/b-/- mice die of starvation within the first two weeks of life. This is due to a localized disintegration of the dorsal tongue epithelium, which results in the build up of a plaque of cell debris that severely impairs feeding. However, approximately 25% of MK6a/b-/- mice survive to adulthood. Remarkably, the surviving MK6a/b-/- mice have normal hair and nails. To our surprise, we discovered MK6 staining both in the hair follicle and the nail bed of MK6a/b-/- mice, indicating the presence of a third MK6 gene. We cloned this previously unknown murine keratin gene and found it to be highly homologous to human K6hf, which is expressed in hair follicles. We therefore termed this gene MK6 hair follicle (MK6hf). The presence of MK6hf in the MK6a/b-/- follicles and nails offers an explanation for the absence of hair and nail defects in MK6a/b-/- animals. PMID- 11489920 TI - Role of phosphatidylinositol 3-kinase and Rab5 effectors in phagosomal biogenesis and mycobacterial phagosome maturation arrest. AB - Phagosomal biogenesis is a fundamental biological process of particular significance for the function of phagocytic and antigen-presenting cells. The precise mechanisms governing maturation of phagosomes into phagolysosomes are not completely understood. Here, we applied the property of pathogenic mycobacteria to cause phagosome maturation arrest in infected macrophages as a tool to dissect critical steps in phagosomal biogenesis. We report the requirement for 3 phosphoinositides and acquisition of Rab5 effector early endosome autoantigen (EEA1) as essential molecular events necessary for phagosomal maturation. Unlike the model phagosomes containing latex beads, which transiently recruited EEA1, mycobacterial phagosomes excluded this regulator of vesicular trafficking that controls membrane tethering and fusion processes within the endosomal pathway and is recruited to endosomal membranes via binding to phosphatidylinositol 3 phosphate (PtdIns[3]P). Inhibitors of phosphatidylinositol 3'(OH)-kinase (PI-3K) activity diminished EEA1 recruitment to newly formed latex bead phagosomes and blocked phagosomal acquisition of late endocytic properties, indicating that generation of PtdIns(3)P plays a role in phagosomal maturation. Microinjection into macrophages of antibodies against EEA1 and the PI-3K hVPS34 reduced acquisition of late endocytic markers by latex bead phagosomes, demonstrating an essential role of these Rab5 effectors in phagosomal biogenesis. The mechanism of EEA1 exclusion from mycobacterial phagosomes was investigated using mycobacterial products. Coating of latex beads with the major mycobacterial cell envelope glycosylated phosphatidylinositol lipoarabinomannan isolated from the virulent Mycobacterium tuberculosis H37Rv, inhibited recruitment of EEA1 to latex bead phagosomes, and diminished their maturation. These findings define the generation of phosphatidylinositol 3-phosphate and EEA1 recruitment as: (a) important regulatory events in phagosomal maturation and (b) critical molecular targets affected by M. tuberculosis. This study also identifies mycobacterial phosphoinositides as products with specialized toxic properties, interfering with discrete trafficking stages in phagosomal maturation. PMID- 11489921 TI - High resolution mapping of mast cell membranes reveals primary and secondary domains of Fc(epsilon)RI and LAT. AB - In mast cells, cross-linking the high-affinity IgE receptor (Fc(epsilon)RI) initiates the Lyn-mediated phosphorylation of receptor ITAMs, forming phospho ITAM binding sites for Syk. Previous immunogold labeling of membrane sheets showed that resting Fc(epsilon)RI colocalize loosely with Lyn, whereas cross linked Fc(epsilon)RI redistribute into specialized domains (osmiophilic patches) that exclude Lyn, accumulate Syk, and are often bordered by coated pits. Here, the distribution of Fc(epsilon)RI beta is mapped relative to linker for activation of T cells (LAT), Grb2-binding protein 2 (Gab2), two PLCgamma isoforms, and the p85 subunit of phosphatidylinositol 3-kinase (PI3-kinase), all implicated in the remodeling of membrane inositol phospholipids. Before activation, PLCgamma1 and Gab2 are not strongly membrane associated, LAT occurs in small membrane clusters separate from receptor, and PLCgamma2, that coprecipitates with LAT, occurs in clusters and along cytoskeletal cables. After activation, PLCgamma2, Gab2, and a portion of p85 colocalize with Fc(epsilon)RI beta in osmiophilic patches. LAT clusters enlarge within 30 s of receptor activation, forming elongated complexes that can intersect osmiophilic patches without mixing. PLCgamma1 and another portion of p85 associate preferentially with activated LAT. Supporting multiple distributions of PI3-kinase, Fc(epsilon)RI cross-linking increases PI3-kinase activity in anti-LAT, anti Fc(epsilon)RIbeta, and anti-Gab2 immune complexes. We propose that activated mast cells propagate signals from primary domains organized around Fc(epsilon)RIbeta and from secondary domains, including one organized around LAT. PMID- 11489922 TI - Coordinated expression of matrix Gla protein is required during endochondral ossification for chondrocyte survival. AB - Matrix Gla protein (MGP) is a 14-kD extracellular matrix protein of the mineral binding Gla protein family. Studies of MGP-deficient mice suggest that MGP is an inhibitor of extracellular matrix calcification in arteries and the epiphyseal growth plate. In the mammalian growth plate, MGP is expressed by proliferative and late hypertrophic chondrocytes, but not by the intervening chondrocytes. To investigate the functional significance of this biphasic expression pattern, we used the ATDC5 mouse chondrogenic cell line. We found that after induction of the cell line with insulin, the differentiating chondrocytes express MGP in a stage specific biphasic manner as in vivo. Treatment of the ATDC5 cultures with MGP antiserum during the proliferative phase leads to their apoptosis before maturation, whereas treatment during the hypertrophic phase has no effect on chondrocyte viability or mineralization. After stable transfection of ATDC5 cells with inducible sense or antisense MGP cDNA constructs, we found that overexpression of MGP in maturing chondrocytes and underexpression of MGP in proliferative and hypertrophic chondrocytes induced apoptosis. However, overexpression of MGP during the hypertrophic phase has no effect on chondrocyte viability, but it does reduce mineralization. This work suggests that coordinated levels of MGP are required for chondrocyte differentiation and matrix mineralization. PMID- 11489923 TI - T cell homeostasis and systemic autoimmunity. PMID- 11489924 TI - Molecular properties and involvement of heparanase in cancer metastasis and angiogenesis. PMID- 11489925 TI - Heparan sulfate proteoglycans: heavy hitters in the angiogenesis arena. PMID- 11489926 TI - Heparan sulfate: growth control with a restricted sequence menu. PMID- 11489927 TI - Indispensable tools: embryonic stem cells yield insights into the human heart. PMID- 11489928 TI - Statins' benefits begin to sprout. PMID- 11489929 TI - Advances in hepatocyte transplantation: a myth becomes reality. PMID- 11489930 TI - Insulin-induced cortical actin remodeling promotes GLUT4 insertion at muscle cell membrane ruffles. AB - Insulin stimulates glucose uptake by recruiting glucose transporter 4 (GLUT4) from an intracellular compartment to the cell surface; this phenomenon is defective in type 2 diabetes. Here we examine the involvement of actin filaments in GLUT4 translocation and their possible defects in insulin resistance, using L6 myotubes expressing myc-tagged GLUT4. Insulin caused membrane ruffling, a dynamic distortion of the myotube dorsal surface. Fluorescence microscopy and immunogold staining of surface GLUT4myc coupled to backscatter electron microscopy revealed a high density of this protein in membrane ruffles. The t-SNAREs syntaxin4 and SNAP-23 were also abundant in these regions. Below the membrane, GLUT4 and the vesicular protein VAMP2, but not VAMP3, colocalized with the actin structures supporting the membrane ruffles. GLUT4myc externalization and membrane ruffles were reduced by jasplakinolide and by swinholide-A, drugs that affect actin filament stability and prevent actin branching, respectively. Insulin resistance generated by prolonged (24 hours) exposure of myotubes to high glucose and insulin diminished the acute insulin-dependent remodeling of cortical actin and GLUT4myc translocation, reminiscent of the effect of swinholide-A. We propose that GLUT4 vesicle incorporation into the plasma membrane involves insulin dependent cortical actin remodeling and that defective actin remodeling contributes to insulin resistance. PMID- 11489931 TI - Loss of p27(Kip1) enhances the transplantation efficiency of hepatocytes transferred into diseased livers. AB - p27(Kip1) is an important regulator of cyclin-dependent kinases. Studies with p27 knockout mice have revealed abnormalities in proliferation and differentiation of multiple cell types. Here we show that primary hepatocytes isolated from livers of adult p27 knockout mice exhibit higher levels of DNA synthesis activity in culture than do wild-type cells. Interestingly, we found that, compared with control hepatocytes, p27 knockout hepatocytes proliferate better after transplantation into diseased livers to reverse liver failure. These results reveal an aspect of p27 that could be used to benefit cell-based therapy. PMID- 11489932 TI - HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. AB - HMG-CoA reductase inhibitors (statins) have been developed as lipid-lowering drugs and are well established to reduce morbidity and mortality from coronary artery disease. Here we demonstrate that statins potently augment endothelial progenitor cell differentiation in mononuclear cells and CD34-positive hematopoietic stem cells isolated from peripheral blood. Moreover, treatment of mice with statins increased c-kit(+)/Sca-1(+)--positive hematopoietic stem cells in the bone marrow and further elevated the number of differentiated endothelial progenitor cells (EPCs). Statins induce EPC differentiation via the PI 3 kinase/Akt (PI3K/Akt) pathway as demonstrated by the inhibitory effect of pharmacological PI3K blockers or overexpression of a dominant negative Akt construct. Similarly, the potent angiogenic growth factor VEGF requires Akt to augment EPC numbers, suggesting an essential role for Akt in regulating hematopoietic progenitor cell differentiation. Given that statins are at least as potent as VEGF in increasing EPC differentiation, augmentation of circulating EPC might importantly contribute to the well-established beneficial effects of statins in patients with coronary artery disease. PMID- 11489933 TI - HMG-CoA reductase inhibitor mobilizes bone marrow--derived endothelial progenitor cells. AB - Endothelial progenitor cells (EPCs) have been isolated from circulating mononuclear cells in peripheral blood and shown to incorporate into foci of neovascularization, consistent with postnatal vasculogenesis. These circulating EPCs are derived from bone marrow and are mobilized endogenously in response to tissue ischemia or exogenously by cytokine stimulation. We show here, using a chemotaxis assay of bone marrow mononuclear cells in vitro and EPC culture assay of peripheral blood from simvastatin-treated animals in vivo, that the HMG-CoA reductase inhibitor, simvastatin, augments the circulating population of EPCs. Direct evidence that this increased pool of circulating EPCs originates from bone marrow and may enhance neovascularization was demonstrated in simvastatin-treated mice transplanted with bone marrow from transgenic donors expressing beta galactosidase transcriptionally regulated by the endothelial cell-specific Tie-2 promoter. The role of Akt signaling in mediating effects of statin on EPCs is suggested by the observation that simvastatin rapidly activates Akt protein kinase in EPCs, enhancing proliferative and migratory activities and cell survival. Furthermore, dominant negative Akt overexpression leads to functional blocking of EPC bioactivity. These findings establish that augmented mobilization of bone marrow-derived EPCs through stimulation of the Akt signaling pathway constitutes a novel function for HMG-CoA reductase inhibitors. PMID- 11489934 TI - Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes. AB - The study of human cardiac tissue development is hampered by the lack of a suitable in vitro model. We describe the phenotypic properties of cardiomyocytes derived from human embryonic stem (ES) cells. Human ES cells were cultivated in suspension and plated to form aggregates termed embryoid bodies (EBs). Spontaneously contracting areas appeared in 8.1% of the EBs. Cells from the spontaneously contracting areas within EBs were stained positively with anti cardiac myosin heavy chain, anti--alpha-actinin, anti-desmin, anti--cardiac troponin I (anti-cTnI), and anti-ANP antibodies. Electron microscopy revealed varying degrees of myofibrillar organization, consistent with early-stage cardiomyocytes. RT-PCR studies demonstrated the expression of several cardiac specific genes and transcription factors. Extracellular electrograms were characterized by a sharp component lasting 30 +/- 25 milliseconds, followed by a slow component of 347 +/- 120 milliseconds. Intracellular Ca(2+) transients displayed a sharp rise lasting 130 +/- 27 milliseconds and a relaxation component lasting 200--300 milliseconds. Positive and negative chronotropic effects were induced by application of isoproterenol and carbamylcholine, respectively. In conclusion, the human ES cell--derived cardiomyocytes displayed structural and functional properties of early-stage cardiomyocytes. Establishment of this unique differentiation system may have significant impact on the study of early human cardiac differentiation, functional genomics, pharmacological testing, cell therapy, and tissue engineering. PMID- 11489935 TI - Conjugation of a self-antigen to papillomavirus-like particles allows for efficient induction of protective autoantibodies. AB - High avidity and long-lasting autoantibodies to a self-polypeptide (TNF-alpha) were generated after parenteral vaccination of mice with low doses of virus-like particle-based (VLP-based) vaccines that were constructed by linking mouse TNF alpha peptides to the surface of papillomavirus VLPs. High-titer autoantibodies were induced with or without coadministration of potent conventional adjuvants, but were enhanced by coadministration of CFA. Compared with immunization with the fusion protein alone, attachment to VLPs increased autoantibody titers 1,000 fold. A comparison of Ab responses against the self (TNF-alpha) and foreign components of the fusion protein showed that VLP conjugation abrogated the ability of the humoral immune system to distinguish between self and foreign. Similar levels of IgM were detected to self and foreign epitopes regardless of the assembly state of the antigen, suggesting that conjugation of self-peptides to VLPs promotes survival or expansion of mature autoreactive B cells. In a mouse model, vaccination with conjugated particles inhibited development of type II collagen-induced arthritis. Together, these results suggest a potentially flexible method to efficiently generate autoantibodies against specific self proteins that mediate arthritis and other diseases. PMID- 11489936 TI - TNF-alpha mediates SDF-1 alpha-induced NF-kappa B activation and cytotoxic effects in primary astrocytes. AB - Stromal-derived cell factor-1 alpha (SDF-1 alpha; CXCL12) and its receptor, CXCR4, are constitutively expressed on neuroepithelial cells and are believed to be involved in both development and pathological processes, such as AIDS associated neurologic disorders. Here, we demonstrate that SDF-1 alpha activates NF-kappa B, stimulates production of chemokines and cytokines, and induces cell death in primary astrocytes, effects that depend on ongoing secretion of TNF alpha. SDF-1 alpha upregulated TNF-alpha mRNA and protein secretion, as well as TNF receptor 2 expression. TNF-alpha treatment mimicked SDF-1 alpha induction of NF-kappa B, IL-1 alpha/beta, and RANTES, as well as cell death; neutralizing antibodies against TNF-alpha opposed these responses. We also found that SDF-1 alpha activated Erk1 and Erk2 (Erk1/2) MAPK in a biphasic fashion. Early Erk1/2 activation was stimulated directly by SDF-1 alpha and late activation was mediated by TNF-alpha. PD98059 suppression of early Erk1/2 activation correlated with reduction of SDF-1 alpha-induced TNF-alpha expression. Late Erk1/2 activation was involved in TNF-alpha-stimulated NF-kappa B activation and cytokine induction. SDF-1 alpha was induced in reactive CXCR4-positive astrocytes near axotomized spinal cord motor neurons, consistent with autocrine SDF-1/CXCR4 signaling in these cells. We propose that these novel effects of SDF-1 alpha are relevant to the pathogenic and developmental roles of SDF-1 alpha in the CNS. PMID- 11489937 TI - Prevention of fat-induced insulin resistance by salicylate. AB - Insulin resistance is a major factor in the pathogenesis of type 2 diabetes and may involve fat-induced activation of a serine kinase cascade involving IKK-beta. To test this hypothesis, we first examined insulin action and signaling in awake rats during hyperinsulinemic-euglycemic clamps after a lipid infusion with or without pretreatment with salicylate, a known inhibitor of IKK-beta. Whole-body glucose uptake and metabolism were estimated using [3-(3)H]glucose infusion, and glucose uptake in individual tissues was estimated using [1-(14)C]2-deoxyglucose injection during the clamp. Here we show that lipid infusion decreased insulin stimulated glucose uptake and activation of IRS-1-associated PI 3-kinase in skeletal muscle but that salicylate pretreatment prevented these lipid-induced effects. To examine the mechanism of salicylate action, we studied the effects of lipid infusion on insulin action and signaling during the clamp in awake mice lacking IKK-beta. Unlike the response in wild-type mice, IKK-beta knockout mice did not exhibit altered skeletal muscle insulin signaling and action following lipid infusion. In summary, high-dose salicylate and inactivation of IKK-beta prevent fat-induced insulin resistance in skeletal muscle by blocking fat-induced defects in insulin signaling and action and represent a potentially novel class of therapeutic agents for type 2 diabetes. PMID- 11489938 TI - Avoidance of stimulation improves engraftment of cultured and retrovirally transduced hematopoietic cells in primates. AB - Recent reports suggest that cells in active cell cycle have an engraftment defect compared with quiescent cells. We used nonhuman primates to investigate this finding, which has direct implications for clinical transplantation and gene therapy applications. Transfer of rhesus CD34(+) cells to culture in stem cell factor (SCF) on the CH-296 fibronectin fragment (FN) after 4 days of culture in stimulatory cytokines maintained cell viability but decreased cycling. Using retroviral marking with two different gene transfer vectors, we compared the engraftment potential of cytokine-stimulated cells versus those transferred to nonstimulatory conditions (SCF on FN alone) before reinfusion. In vivo competitive repopulation studies showed that the level of marking originating from the cells continued in culture for 2 days with SCF on FN following a 4-day stimulatory transduction was significantly higher than the level of marking coming from cells transduced for 4 days and reinfused without the 2-day culture under nonstimulatory conditions. We observed stable in vivo overall gene marking levels of up to 29%. This approach may allow more efficient engraftment of transduced or ex vivo expanded cells by avoiding active cell cycling at the time of reinfusion. PMID- 11489939 TI - Hyperinsulinism in short-chain L-3-hydroxyacyl-CoA dehydrogenase deficiency reveals the importance of beta-oxidation in insulin secretion. AB - A female infant of nonconsanguineous Indian parents presented at 4 months with a hypoglycemic convulsion. Further episodes of hypoketotic hypoglycemia were associated with inappropriately elevated plasma insulin concentrations. However, unlike other children with hyperinsulinism, this patient had a persistently elevated blood spot hydroxybutyrylcarnitine concentration when fed, as well as when fasted. Measurement of the activity of L-3-hydroxyacyl-CoA dehydrogenase in cultured skin fibroblasts with acetoacetyl-CoA substrate showed reduced activity. In fibroblast mitochondria, the activity was less than 5% that of controls. Sequencing of the short-chain L-3-hydroxyacyl-CoA dehydrogenase (SCHAD) genomic DNA from the fibroblasts showed a homozygous mutation (C773T) changing proline to leucine at amino acid 258. Analysis of blood from the parents showed they were heterozygous for this mutation. Western blot studies showed undetectable levels of immunoreactive SCHAD protein in the child's fibroblasts. Expression studies showed that the P258L enzyme had no catalytic activity. We conclude that C773T is a disease-causing SCHAD mutation. This is the first defect in fatty acid beta oxidation that has been associated with hyperinsulinism and raises interesting questions about the ways in which changes in fatty acid and ketone body metabolism modulate insulin secretion by the beta cell. The patient's hyperinsulinism was easily controlled with diazoxide and chlorothiazide. PMID- 11489940 TI - Impaired JAK-STAT signal transduction contributes to growth hormone resistance in chronic uremia. AB - Chronic renal failure (CRF) is associated with resistance to the growth-promoting and anabolic actions of growth hormone (GH). In rats with CRF induced by partial renal ablation, 7 days of GH treatment had a diminished effect on weight gain and hepatic IGF-1 and IGFBP-1 mRNA levels, compared with sham-operated pair-fed controls. To assess whether GH resistance might be due to altered signal transduction, activation of the JAK-STAT pathway was studied 10 or 15 minutes after intravenous injection of 5 mg/kg GH or vehicle. Hepatic GH receptor (GHR) mRNA levels were significantly decreased in CRF, but GHR protein abundance and GH binding to microsomal and plasma membranes was unaltered. JAK2, STAT1, STAT3, and STAT5 protein abundance was also unchanged. However, GH-induced tyrosine phosphorylation of JAK2, STAT5, and STAT3 was 75% lower in the CRF animals. Phosphorylated STAT5 and STAT3 were also diminished in nuclear extracts. The expression of the suppressor of cytokine signaling-2 (SOCS-2) was increased twofold in GH-treated CRF animals, and SOCS-3 mRNA levels were elevated by 60% in CRF, independent of GH treatment. In conclusion, CRF causes a postreceptor defect in GH signal transduction characterized by impaired phosphorylation and nuclear translocation of GH-activated STAT proteins, which is possibly mediated, at least in part, by overexpression of SOCS proteins. PMID- 11489941 TI - Impaired activation of murine platelets lacking G alpha(i2). AB - The intracellular signaling pathways by which G protein-coupled receptors on the platelet surface initiate aggregation, a critical process for hemostasis and thrombosis, are not well understood. In particular, the contribution of the G(i) pathway has not been directly addressed. We have investigated the activation of platelets from mice in which the gene for the predominant platelet G alpha(i) subtype, G alpha(i2), has been disrupted. In intact platelets from G alpha(i2) deficient mice, the inhibition of adenylyl cyclase by ADP was found to be partially impaired compared with wild-type platelets. Moreover, both ADP dependent platelet aggregation and the activation of the integrin alpha IIb beta 3 (GPIIb-IIIa) were strongly reduced in platelets from G alpha(i2)-deficient mice. In addition, G alpha(i2)-deficient platelets displayed impaired activation at low thrombin concentrations. This defect was mimicked by blocking the adenylyl cyclase--coupled platelet ADP receptor (P2Y(12)) on wild-type platelets with a selective antagonist. These observations suggest that G alpha(i2) is involved in the inhibition of platelet adenylyl cyclase in vivo and is a critical component of the signaling pathway for integrin activation by ADP, resulting in platelet aggregation. In addition, thrombin-dependent activation of mouse platelets is mediated, at least in part, by secreted ADP acting on the G alpha(i2)-linked ADP receptor. PMID- 11489942 TI - Plasma CD14 decreases monocyte responses to LPS by transferring cell-bound LPS to plasma lipoproteins. AB - CD14, a myeloid cell-surface receptor and soluble plasma protein, binds LPS and other microbial molecules and initiates the innate immune response to bacterial invasion. The blood concentration of soluble CD14 (sCD14) increases during the systemic response to infection. Although high sCD14 blood levels have correlated with increased risk of dying from severe sepsis, sCD14 can diminish cell responses to LPS. We show here that in human serum, sCD14 increases the rate at which cell-bound LPS is released from the monocyte surface and binds to plasma lipoproteins. This enhanced rate of LPS efflux is associated with a significant reduction in the ability of monocytes to produce cytokines in response to LPS. Serum from septic patients reduced the LPS-monocyte interaction by as much as tenfold, and depletion of sCD14 from the serum restored LPS-monocyte binding and release kinetics to near normal levels. In serum from septic patients, monocyte bound LPS also moved more rapidly into lipoproteins, which completely neutralized the biologic activity of the LPS that bound to them. In human plasma, sCD14 thus diminishes monocyte responses to LPS by transferring cell-bound LPS to lipoproteins. Stress-related increases in plasma sCD14 levels may help prevent inflammatory responses within the blood. PMID- 11489943 TI - NTB-A [correction of GNTB-A], a novel SH2D1A-associated surface molecule contributing to the inability of natural killer cells to kill Epstein-Barr virus infected B cells in X-linked lymphoproliferative disease. AB - In humans, natural killer (NK) cell function is regulated by a series of receptors and coreceptors with either triggering or inhibitory activity. Here we describe a novel 60-kD glycoprotein, termed NTB-A, that is expressed by all human NK, T, and B lymphocytes. Monoclonal antibody (mAb)-mediated cross-linking of NTB A results in the induction of NK-mediated cytotoxicity. Similar to 2B4 (CD244) functioning as a coreceptor in the NK cell activation, NTB-A also triggers cytolytic activity only in NK cells expressing high surface densities of natural cytotoxicity receptors. This suggests that also NTB-A may function as a coreceptor in the process of NK cell activation. Molecular cloning of the cDNA coding for NTB-A molecule revealed a novel member of the immunoglobulin superfamily belonging to the CD2 subfamily. NTB-A is characterized, in its extracellular portion, by a distal V-type and a proximal C2-type domain and by a cytoplasmic portion containing three tyrosine-based motifs. NTB-A undergoes tyrosine phosphorylation and associates with the Src homology 2 domain-containing protein (SH2D1A) as well as with SH2 domain-containing phosphatases (SHPs). Importantly, analysis of NK cells derived from patients with X-linked lymphoproliferative disease (XLP) showed that the lack of SH2D1A protein profoundly affects the function not only of 2B4 but also of NTB-A. Thus, in XLP NK cells, NTB-A mediates inhibitory rather than activating signals. These inhibitory signals are induced by the interaction of NTB-A with still undefined ligands expressed on Epstein-Barr virus (EBV)-infected target cells. Moreover, mAb-mediated masking of NTB-A can partially revert this inhibitory effect while a maximal recovery of target cell lysis can be obtained when both 2B4 and NTB-A are simultaneously masked. Thus, the altered function of NTB-A appears to play an important role in the inability of XLP-NK cells to kill EBV-infected target cells. PMID- 11489944 TI - Transplanted long-term cultured pre-BI cells expressing calpastatin are resistant to B cell receptor-induced apoptosis. AB - Long-term cultured pre-B cells are able to differentiate into immunoglobulin (Ig)M-positive B cells (IgM(+) cells) when transplanted into severe combined immunodeficient (SCID) mice. Based on previous studies, here we report the development of a reconstitution assay in nonobese diabetic/SCID (NOD/SCID) mice using pre-B cells, which allows us to study the role of calpains (calcium activated endopeptidases) during B cell development as well as in B cell clonal deletion. Using this model, we show that calpastatin (the natural inhibitor of calpains) inhibits B cell receptor-induced apoptosis in IgM(+) cells derived from transplanted mice. We thus hypothesize an important function for calpain in sculpting the B cell repertoire. PMID- 11489946 TI - Phosphoinositide 3-kinase-dependent membrane recruitment of p62(dok) is essential for its negative effect on mitogen-activated protein (MAP) kinase activation. AB - A major pathway by which growth factors, such as platelet-derived growth factor (PDGF), regulate cell proliferation is via the receptor tyrosine kinase/Ras/mitogen-activated protein kinase (MAPK) signaling cascade. The output of this pathway is subjected to tight regulation of both positive and negative regulators. One such regulator is p62(dok), the prototype of a newly identified family of adaptor proteins. We recently provided evidence, through the use of p62(dok)-deficient cells, that p62(dok) acts as a negative regulator of growth factor-induced cell proliferation and the Ras/MAPK pathway. We show here that reintroduction of p62(dok) into p62(dok)-(/)- cells can suppress the increased cell proliferation and prolonged MAPK activity seen in these cells, and that plasma membrane recruitment of p62(dok) is essential for its function. We also show that the PDGF-triggered plasma membrane translocation of p62(dok) requires activation of phosphoinositide 3-kinase (PI3-kinase) and binding of its pleckstrin homology (PH) domain to 3'-phosphorylated phosphoinositides. Furthermore, we demonstrate that p62(dok) can exert its negative effect on the PDGFR/MAPK pathway independently of its ability to associate with RasGAP and Nck. We conclude that p62(dok) functions as a negative regulator of the PDGFR/Ras/MAPK signaling pathway through a mechanism involving PI3-kinase-dependent recruitment of p62(dok) to the plasma membrane. PMID- 11489945 TI - Epstein-Barr virus latent membrane protein 2A (LMP2A) employs the SLP-65 signaling module. AB - In latently infected B lymphocytes, the Epstein-Barr virus (EBV) suppresses signal transduction from the antigen receptor through expression of the integral latent membrane protein 2A (LMP2A). At the same time, LMP2A triggers B cell survival by a yet uncharacterized maintenance signal that is normally provided by the antigen receptor. The molecular mechanisms are unknown as LMP2A-regulated signaling cascades have not been described so far. Using a novel mouse model we have identified the intracellular adaptor protein Src homology 2 (SH2) domain containing leukocyte protein (SLP)-65 as a critical downstream effector of LMP2A in vivo. Biochemical analysis of the underlying signaling pathways revealed that EBV infection causes constitutive tyrosine phosphorylation of one of the two SLP 65 isoforms and complex formation between SLP-65 and the protooncoprotein CrkL (CT10 regulator of kinase like). This leads to antigen receptor-independent phosphorylation of Cbl (Casitas B lineage lymphoma) and C3G. In contrast, phospholipase C-gamma2 (PLC-gamma2) activation is completely blocked. Our data show that in order to establish a latent EBV infection, LMP2A selectively activates or represses SLP-65-regulated signaling pathways. PMID- 11489947 TI - p62(dok), a negative regulator of Ras and mitogen-activated protein kinase (MAPK) activity, opposes leukemogenesis by p210(bcr-abl). AB - p62(dok) has been identified as a substrate of many oncogenic tyrosine kinases such as the chronic myelogenous leukemia (CML) chimeric p210(bcr-abl) oncoprotein. It is also phosphorylated upon activation of many receptors and cytoplamic tyrosine kinases. However, the biological functions of p62(dok) in normal cell signaling as well as in p210(bcr-abl) leukemogenesis are as yet not fully understood. Here we show, in hemopoietic and nonhemopoietic cells derived from p62(dok)-(/)- mice, that the loss of p62(dok) results in increased cell proliferation upon growth factor treatment. Moreover, Ras and mitogen-activated protein kinase (MAPK) activation is markedly sustained in p62(dok)-(/)- cells after the removal of growth factor. However, p62(dok) inactivation does not affect DNA damage and growth factor deprivation-induced apoptosis. Furthermore, p62(dok) inactivation causes a significant shortening in the latency of the fatal myeloproliferative disease induced by retroviral-mediated transduction of p210(bcr-abl) in bone marrow cells. These data indicate that p62(dok) acts as a negative regulator of growth factor-induced cell proliferation, at least in part through downregulating Ras/MAPK signaling pathway, and that p62(dok) can oppose leukemogenesis by p210(bcr-abl). PMID- 11489948 TI - Point mutation in essential genes with loss or mutation of the second allele: relevance to the retention of tumor-specific antigens. AB - Antigens that are tumor specific yet retained by tumor cells despite tumor progression offer stable and specific targets for immunologic and possibly other therapeutic interventions. Therefore, we have studied two CD4(+) T cell recognized tumor-specific antigens that were retained during evolution of two ultraviolet-light-induced murine cancers to more aggressive growth. The antigens are ribosomal proteins altered by somatic tumor-specific point mutations, and the progressor (PRO) variants lack the corresponding normal alleles. In the first tumor, 6132A-PRO, the antigen is encoded by a point-mutated L9 ribosomal protein gene. The tumor lacks the normal L9 allele because of an interstitial deletion from chromosome 5. In the second tumor, 6139B-PRO, both alleles of the L26 gene have point mutations, and each encodes a different tumor-specific CD4(+) T cell recognized antigen. Thus, for both L9 and L26 genes, we observe "two hit" kinetics commonly observed in genes suppressing tumor growth. Indeed, reintroduction of the lost wild-type L9 allele into the 6132A-PRO variant suppressed the growth of the tumor cells in vivo. Since both L9 and L26 encode proteins essential for ribosomal biogenesis, complete loss of the tumor-specific target antigens in the absence of a normal allele would abrogate tumor growth. PMID- 11489949 TI - A secreted chemokine binding protein encoded by murine gammaherpesvirus-68 is necessary for the establishment of a normal latent load. AB - Herpesviruses encode a variety of proteins with the potential to disrupt chemokine signaling, and hence immune organization. However, little is known of how these might function in vivo. The B cell-tropic murine gammaherpesvirus-68 (MHV-68) is related to the Kaposi's sarcoma-associated herpesvirus (KSHV), but whereas KSHV expresses small chemokine homologues, MHV-68 encodes a broad spectrum chemokine binding protein (M3). Here we have analyzed the effect on viral pathogenesis of a targeted disruption of the M3 gene. After intranasal infection, an M3 deficiency had surprisingly little effect on lytic cycle replication in the respiratory tract or the initial spread of virus to lymphoid tissues. However, the amplification of latently infected B cells in the spleen that normally drives MHV-68-induced infectious mononucleosis failed to occur. Thus, there was a marked reduction in latent virus recoverable by in vitro reactivation, latency-associated viral tRNA transcripts detectable by in situ hybridization, total viral DNA load, and virus-driven B cell activation. In vivo CD8(+) T cell depletion largely reversed this deficiency, suggesting that the chemokine neutralization afforded by M3 may function to block effective CD8(+) T cell recruitment into lymphoid tissue during the expansion of latently infected B cell numbers. In the absence of M3, MHV-68 was unable to establish a normal latent load. PMID- 11489950 TI - CD1-restricted NK T cells protect nonobese diabetic mice from developing diabetes. AB - NK T cells are a unique subset of T cells that recognize lipid antigens presented by CD1d. After activation, NK T cells promptly produce large amounts of cytokines, which may modulate the upcoming immune responses. Previous studies have documented an association between decreased numbers of NK T cells and the progression of some autoimmune diseases, suggesting that NK T cells may control the development of autoimmune diseases. To investigate the role of NK T cells in autoimmune diabetes, we crossed CD1 knockout (CD1KO) mutation onto the nonobese diabetic (NOD) genetic background. We found that male CD1KO NOD mice exhibited significantly higher incidence and earlier onset of diabetes compared with the heterozygous controls. The diabetic frequencies in female mice showed a similar pattern; however, the differences were less profound between female CD1KO and control mice. Early treatment of NOD mice with alpha-galactosylceramide, a potent NK T cell activator, reduced the severity of autoimmune diabetes in a CD1 dependent manner. Our results not only suggest a protective role of CD1 restricted NK T cells in autoimmune diabetes but also reveal a causative link between the deficiency of NK T cells and the induction of insulin-dependent diabetes mellitus. PMID- 11489951 TI - Genetic influences on the end-stage effector phase of arthritis. AB - K/BxN T cell receptor transgenic mice are a model of inflammatory arthritis, most similar to rheumatoid arthritis, that is critically dependent on both T and B lymphocytes. Transfer of serum, or just immunoglobulins, from arthritic K/BxN animals into healthy recipients provokes arthritis efficiently, rapidly, and with high penetrance. We have explored the genetic heterogeneity in the response to serum transfer, thereby focussing on the end-stage effector phase of arthritis, leap-frogging the initiating events. Inbred mouse strains showed clear variability in their responses. A few were entirely refractory to disease induction, and those which did develop disease exhibited a range of severities. F1 analyses suggested that in most cases susceptibility was controlled in a polygenic additive fashion. One responder/nonresponder pair (C57Bl/6 x NOD) was studied in detail via a genome scan of F2 mice; supplementary information was provided by the examination of knock-out and congenic strains. Two genomic regions that are major, additive determinants of the rapidity and severity of K/BxN serum-transferred arthritis were highlighted. Concerning the first region, on proximal chromosome (chr)2, candidate assignment to the complement gene C5 was confirmed by both strain segregation analysis and functional data. Concerning the second, on distal chr1, coinciding with the Sle1 locus implicated in susceptibility to lupus-like autoimmune disease, a contribution by the fcgr2 candidate gene was excluded. Two other regions, on chr12 and chr18 may also contribute to susceptibility to serum-transferred arthritis, albeit to a more limited degree. The contributions of these loci are additive, but gene dosage effects at the C5 locus are such that it largely dominates. The clarity of these results argues that our focus on the terminal effector phase of arthritis in the K/BxN model will bear fruit. PMID- 11489952 TI - Toward a defined anti-Leishmania vaccine targeting vector antigens: characterization of a protective salivary protein. AB - Leishmania parasites are transmitted to their vertebrate hosts by infected phlebotomine sand fly bites. Sand fly saliva is known to enhance Leishmania infection, while immunity to the saliva protects against infection as determined by coinoculation of parasites with vector salivary gland homogenates (SGHs) or by infected sand fly bites (Kamhawi, S., Y. Belkaid, G. Modi, E. Rowton, and D. Sacks. 2000. Science. 290:1351-1354). We have now characterized nine salivary proteins of Phlebotomus papatasi, the vector of Leishmania major. One of these salivary proteins, extracted from SDS gels and having an apparent mol wt of 15 kD, was able to protect vaccinated mice challenged with parasites plus SGH. A DNA vaccine containing the cDNA for the predominant 15-kD protein (named SP15) provided this same protection. Protection lasted at least 3 mo after immunization. The vaccine produced both intense humoral and delayed-type hypersensitivity (DTH) reactions. B cell-deficient mice immunized with the SP15 plasmid vaccine successfully controlled Leishmania infection when injected with Leishmania plus SGH. These results indicate that DTH response against saliva provides most or all of the protective effects of this vaccine and that salivary gland proteins or their cDNAs are viable vaccine targets against leishmaniasis. PMID- 11489953 TI - A critical role for interleukin 18 in primary and memory effector responses to Listeria monocytogenes that extends beyond its effects on Interferon gamma production. AB - The stimulation of interferon (IFN)-gamma by interleukin (IL)-12 has been shown to provide protection from intracellular pathogens such as Listeria monocytogenes. Tumor necrosis factor (TNF) is also a major player in the resolution of Listeria infections and is suggested to have more global effects than can be explained by the induction of IFN-gamma alone. Since IL-18 synergizes with IL-12 to induce IFN-gamma production by natural killer and T helper (Th)1 cells, we determined its role in responses to Listeria. IL-18 appeared to be even more potent than either IL-12 or IFN-gamma for protection against this pathogen and IL-18 enhanced bacterial clearance in the complete absence of IFN-gamma. Indeed IL-18 was comparable to TNF in its ability to resolve the infection and showed a lowered protective capacity in the absence of TNF. Moreover, IL-18 induced macrophages to secrete both TNF and nitric oxide after a Listeria infection. IL-18 was also essential for optimal IFN-gamma production by antigen specific T cells. Therefore, IL-18 operates via its effects on both the innate immune response, including macrophages, as well as on Th1 cells, to protect against Listeria. PMID- 11489954 TI - Interleukin (IL)-18 promotes the development of chronic gastrointestinal helminth infection by downregulating IL-13. AB - Expulsion of the gastrointestinal nematode Trichuris muris is mediated by a T helper (Th) 2 type response involving interleukin (IL)-4 and IL-13. Here we show that Th1 response-associated susceptibility involves prior activation of IL-18 and caspase-1 followed by IL-12 and interferon (IFN)-gamma in the intestine. IL 18-deficient mice are highly resistant to chronic T. muris infection and in vivo treatment of normal mice with recombinant (r)IL-18 suppresses IL-13 and IL-4 secretion but does not affect IFN-gamma. In vivo treatment of T. muris-infected IFN-gamma-deficient mice with rIL-18 demonstrated that the inhibitory effect of IL-18 on IL-13 secretion is independent of IFN-gamma. Hence, IL-18 does not function as an IFN-gamma-inducing cytokine during chronic T. muris infection but rather as a direct regulator of Th2 cytokines. These results provide the first demonstration of the critical role of IL-18 in regulating Th cell responses during gastrointestinal nematode infection. PMID- 11489955 TI - Quantitative regulation of class switch recombination by switch region transcription. AB - The isotype specificity of immunoglobulin (Ig) class switching is regulated by a cytokine which induces transcription of a specific switch (S) region, giving rise to so-called germline transcripts. Although previous studies have demonstrated that germline transcription of an S region is required for class switch recombination (CSR) of that particular S region, it has not been shown whether the level of S region transcription affects the efficiency of CSR. We addressed this question by using an artificial DNA construct containing a constitutively transcribed mu switch (Smu) region and an alpha switch (Salpha) region driven by a tetracycline-responsive promoter. The construct was introduced into a switch inducible B lymphoma line and the quantitative correlation between Salpha region transcription and class switching efficiency was evaluated. The level of Salpha transcription was linearly correlated with CSR efficiency, reaching a plateau at saturation. On the other hand, we failed to obtain the evidence to support involvement of either RNA-DNA heteroduplex or trans germline transcripts in CSR. Taken together, it is likely that S region transcription and/or transcript processing in situ may be required for CSR. We propose that because of the unusual properties of S region DNA, transcription induces the DNA to transiently be single stranded, permitting secondary structure(s) to form. Such structures may be recognition targets of a putative class switch recombinase. PMID- 11489956 TI - Somatic hypermutation shapes the antibody repertoire of memory B cells in humans. AB - High-affinity antibodies produced by memory B cells differ from antibodies produced in naive B cells in two respects. First, many of these antibodies show somatic hypermutation, and second, the repertoire of antibodies expressed in memory responses is highly selected. To determine whether somatic hypermutation is responsible for the shift in the antibody repertoire during affinity maturation, we analyzed the immunoglobulin lambda light chain (Iglambda) repertoire expressed by naive and antigen-selected memory B cells in humans. We found that the Iglambda repertoire differs between naive and memory B cells and that this shift in the repertoire does not occur in the absence of somatic hypermutation in patients lacking activation-induced cytidine deaminase (AID). Our work suggests that somatic hypermutation makes a significant contribution to shaping the antigen-selected antibody repertoire in humans. PMID- 11489957 TI - Leishmaniasis vaccination: targeting the source of infection. PMID- 11489958 TI - Interleukin 18: tipping the balance towards a T helper cell 1 response. PMID- 11489960 TI - In memoriam Alfred Nisonoff 1923-2001. PMID- 11489961 TI - Cutting edge: T cell migration regulated by CXCR4 chemokine receptor signaling to ZAP-70 tyrosine kinase. AB - Chemokines regulate the homeostatic trafficking of lymphocytes and lymphocyte influx into sites of injury and inflammation. The signaling pathways by which chemokine receptors regulate lymphocyte migration remain incompletely characterized. We demonstrate that Jurkat T cells lacking the ZAP-70 tyrosine kinase exhibit reduced migration in response to the CXCR4 ligand CXCL12 when compared with wild-type Jurkat T cells. Expression of wild-type, but not kinase inactive, ZAP-70 resulted in enhanced migration of ZAP-70-deficient Jurkat T cells. The tyrosine residue at position 292 in the interdomain B region of ZAP-70 exerts a negative regulatory effect on ZAP-70-dependent migration. Stimulation of Jurkat T cells with CXCL12 also resulted in ZAP-70-dependent tyrosine phosphorylation of the Src homology 2 domain-containing leukocyte protein of 76 kDa (SLP-76) adapter protein. Although CXCL12-dependent migration of SLP-76 deficient Jurkat T cells was impaired, re-expression of SLP-76 did not enhance migration. These results suggest a novel function for ZAP-70, but not SLP-76, in CXCR4 chemokine receptor signaling in human T cells. PMID- 11489962 TI - Cutting edge: selective usage of chemokine receptors by plasmacytoid dendritic cells. AB - The existence of dendritic cell (DC) subsets is firmly established, but their trafficking properties are virtually unknown. In this study, we show that myeloid (M-DCs) and plasmacytoid (P-DCs) DCs isolated from human blood differ widely in the capacity to migrate to chemotactic stimuli. The pattern of chemokine receptors expressed by blood M-DCs and P-DCs, with the exception of CCR7, is similar. However, most chemokine receptors of P-DCs, in particular those specific for inflammatory chemokines and classical chemotactic agonists, are not functional in circulating cells. Following maturation induced by CD40 ligation, the receptors for inflammatory chemokines are down-regulated, and CCR7 on P-DCs becomes coupled to migration. The drastically impaired capacity of blood P-DCs to migrate in response to inflammatory chemotactic signals contrasts with the response to lymph node-homing chemokines, indicating a propensity to migrate to secondary lymphoid organs rather than to sites of inflammation. PMID- 11489963 TI - Cutting edge: predominant expression of a novel Ikaros isoform in normal human hemopoiesis. AB - Murine studies implicate Ikaros proteins as regulators of hemopoiesis, particularly in the lymphoid lineages. High homology between murine and human Ikaros suggests that Ikaros expression in the two might be similar. However, initial human studies that focused on leukemia detected novel Ikaros transcripts in patient samples. Thus, novel Ikaros splice forms and DNA nonbinding isoforms were linked with malignancy. We undertook an extensive analysis of normal human Ikaros expression to determine whether novel mRNAs are expressed as proteins and the extent to which these splice variants are unique to leukemia. Here we show that both mRNA and protein for DNA nonbinding Ikaros isoforms and splice variants previously linked to leukemia are expressed in normal human cells. However, our studies identify a new Ikaros isoform not previously described in mouse or human. This isoform is the predominant Ikaros protein in normal human cells, but not in leukemia cell lines. PMID- 11489964 TI - Cutting edge: the nucleotide receptor P2X7 contains multiple protein- and lipid interaction motifs including a potential binding site for bacterial lipopolysaccharide. AB - The nucleotide receptor P2X7 has been shown to modulate LPS-induced macrophage production of numerous inflammatory mediators. Although the C-terminal portion of P2X7 is thought to be essential for multiple receptor functions, little is known regarding the structural motifs that lie within this region. We show here that the P2X7 C-terminal domain contains several apparent protein-protein and protein lipid interaction motifs with potential importance to macrophage signaling and LPS action. Surprisingly, P2X7 also contains a conserved LPS-binding domain. In this report, we demonstrate that peptides derived from this P2X7 sequence bind LPS in vitro. Moreover, these peptides neutralize the ability of LPS to activate the extracellular signal-regulated kinases (ERK1, ERK2) and to promote the degradation of the inhibitor of kappaB-alpha isoform (IkappaB-alpha) in RAW 264.7 macrophages. Collectively, these data suggest that the C-terminal domain of P2X7 may directly coordinate several signal transduction events related to macrophage function and LPS action. PMID- 11489965 TI - Cutting edge: induction of IFN-gamma production but not cytotoxicity by the killer cell Ig-like receptor KIR2DL4 (CD158d) in resting NK cells. AB - Activated NK cells lyse tumor cells and virus-infected cells and produce IFN gamma upon contact with sensitive target cells. The regulation of these effector responses in resting NK cells is not well understood. We now describe a receptor, KIR2DL4, that has the unique property of inducing IFN-gamma production, but not cytotoxicity, by resting NK cells in the absence of cytokines. In contrast, the NK cell-activation receptors CD16 and 2B4 induced cytotoxicity but not IFN-gamma production. The induction by KIR2DL4 of IFN-gamma production by resting NK cells was blocked by an inhibitor of the p38 mitogen-activated protein kinase signaling pathway, in contrast to the IL-2-induced IFN-gamma secretion that was sensitive to inhibition of the extracellular signal-regulated kinase mitogen-activated protein kinase pathway. These results reveal a functional dichotomy (cytokine production vs cytotoxicity) in the response of resting NK cells, as dictated by the signals of individual receptors. PMID- 11489966 TI - Cutting edge: bacterial flagellin activates basolaterally expressed TLR5 to induce epithelial proinflammatory gene expression. AB - Flagellin, the structural component of bacterial flagella, is secreted by pathogenic and commensal bacteria. Flagellin activates proinflammatory gene expression in intestinal epithelia. However, only flagellin that contacts basolateral epithelial surfaces is proinflammatory; apical flagellin has no effect. Pathogenic Salmonella, but not commensal Escherichia coli, translocate flagellin across epithelia, thus activating epithelial proinflammatory gene expression. Investigating how epithelia detect flagellin revealed that cell surface expression of Toll-like receptor 5 (TLR5) conferred NF-kappaB gene expression in response to flagellin. The response depended on both extracellular leucine-rich repeats and intracellular Toll/IL-1R homology region of TLR5 as well as the adaptor protein MyD88. Furthermore, immunolocalization and cell surface selective biotinylation revealed that TLR5 is expressed exclusively on the basolateral surface of intestinal epithelia, thus providing a molecular basis for the polarity of this innate immune response. Thus, detection of flagellin by basolateral TLR5 mediates epithelial-driven inflammatory responses to Salmonella. PMID- 11489967 TI - Cutting edge: expansion and activation of a population of autoreactive marginal zone B cells in a model of estrogen-induced lupus. AB - We have demonstrated previously that 17 beta-estradiol (E2) treatment of BALB/c mice transgenic for the heavy chain of a pathogenic anti-DNA Ab induces a lupus like phenotype with expansion of anti-DNA B cells, elevation of anti-DNA Ab titers, and glomerular immunoglobulin deposition. To understand this loss of B cell tolerance, the effects of E2 on B cell development and activation were examined. A sustained increase in E2 resulted in an altered distribution of B cell subsets, with a diminished transitional population and an increase in marginal zone B cells. Depletion of CD4+ T cells did not abrogate these effects. Furthermore, the B cells that spontaneously secreted anti-DNA Abs displayed a marginal zone phenotype. Thus, a sustained increase in E2 alters B cell development, leading to the survival, expansion, and activation of a population of autoreactive marginal zone B cells implicating this B cell subset in autoimmunity. PMID- 11489968 TI - Role of CD4+ and CD8+ T cells in allorecognition: lessons from corneal transplantation. AB - Corneal transplantation represents an interesting model to investigate the contribution of direct vs indirect Ag recognition pathways to the alloresponse. Corneal allografts are naturally devoid of MHC class II+ APCs. In addition, minor Ag-mismatched corneal grafts are more readily rejected than their MHC-mismatched counterparts. Accordingly, it has been hypothesized that these transplants do not trigger direct T cell alloresponse, but that donor Ags are presented by host APCs, i.e., in an indirect fashion. Here, we have determined the Ag specificity, frequency, and phenotype of T cells activated through direct and indirect pathways in BALB/c mice transplanted orthotopically with fully allogeneic C57BL/6 corneas. In this combination, only 60% of the corneas are rejected, while the remainder enjoy indefinite graft survival. In rejecting mice the T cell response was mediated by two T cell subsets: 1) CD4+ T cells that recognize alloantigens exclusively through indirect pathway and secrete IL-2, and 2) IFN-gamma-producing CD8+ T cells recognizing donor MHC in a direct fashion. Surprisingly, CD8+ T cells activated directly were not required for graft rejection. In nonrejecting mice, no T cell responses were detected. Strikingly, peripheral sensitization to allogeneic MHC molecules in these mice induced acute rejection of corneal grafts. We conclude that only CD4+ T cells activated via indirect allorecognition have the ability to reject allogeneic corneal grafts. Although alloreactive CD8+ T cells are activated via the direct pathway, they are not fully competent and cannot contribute to the rejection unless they receive an additional signal provided by professional APCs in the periphery. PMID- 11489969 TI - Human cytomegalovirus circumvents NF-kappa B dependence in retinal pigment epithelial cells. AB - The human CMV (HCMV) is a persistent virus that may cause severe inflammatory responses especially in immunocompromised hosts. In different cell types, HCMV infection leads to the activation of the pleiotropic transcription factor, NF kappaB, which triggers virus replication but also propagates cell-mediated inflammatory mechanisms that largely depend on PG synthesis. We investigated the interactions of HCMV and the NF-kappaB-dependent PG synthesis pathway in cultures of retinal pigment epithelial (RPE) cells that are known to be infected in HCMV retinitis patients. Unlike in other cell types, HCMV increased neither NF-kappaB activity nor p65 and p105/50 mRNA levels in RPE cells. Both TNF-alpha and phorbol ester 12,0-tetradecanoylphorbol 13-acetate (TPA) enhanced NF-kappaB activity but only TPA increased HCMV replication. Cyclooxygenase-2 expression and PGE2 release was increased by TPA and TNF-alpha but not by HCMV infection. Stimulatory activity of TPA on HCMV replication was suppressed by protein kinase C inhibitors and inhibitors of p42/44 and p38 mitogen-activated protein kinases but not by NF kappaB inhibitors. In conclusion, HCMV circumvents the NF-kappaB route in favor of the protein kinase C-dependent mitogen-activated protein kinase pathway in RPE cells. This virus/host cell interaction might be a mechanism that promotes HCMV persistence in immune-privileged organs such as the eye. PMID- 11489970 TI - Essential role of RelB in germinal center and marginal zone formation and proper expression of homing chemokines. AB - High levels of the Rel/NF-kappaB family member RelB are restricted to specific regions of thymus, lymph nodes, and Peyer's patches. In spleen, RelB is expressed in periarteriolar lymphatic sheaths, germinal centers (GCs), and the marginal zone (MZ). In this study, we report that RelB-deficient (relB(-/-)) mice, in contrast to nfkb1(-/-), but similar to nfkb2(-/-) mice, are unable to form GCs and follicular dendritic cell networks upon Ag challenge in the spleen. RelB is also required for normal organization of the MZ and its population by macrophages and B cells. Reciprocal bone marrow transfers demonstrate that RelB expression in radiation-resistant stromal cells, but not in bone marrow-derived hemopoietic cells, is required for proper formation of GCs, follicular dendritic cell networks, and MZ structures. However, the generation of MZ B cells requires RelB in hemopoietic cells. Expression of TNF ligand/receptor family members is only moderately altered in relB(-/-) splenocytes. In contrast, expression of homing chemokines is strongly reduced in relB(-/-) spleen with particularly low mRNA levels of the chemokine B lymphocyte chemoattractant. Our data indicate that activation of p52-RelB heterodimers in stromal cells downstream of TNF/lymphotoxin is required for normal expression of homing chemokines and proper development of spleen microarchitecture. PMID- 11489971 TI - Signaling through TNF receptor p55 in TNF-alpha-deficient mice alters the CXCL13/CCL19/CCL21 ratio in the spleen and induces maturation and migration of anergic B cells into the B cell follicle. AB - The organization of secondary lymphoid tissues into distinct T and B cell compartments supports proper regulation of an immune response to foreign Ags. In the splenic white pulp, this compartmentalization is also thought to be important in the maintenance of B cell tolerance. Using lymphotoxin-alpha-(LT-alpha)-, TNF alpha-, or TNFRp55-deficient mice, all with disrupted splenic architecture, we tested whether normal T/B segregation and/or intact follicular structure are necessary for the maintenance of anti-dsDNA B cell anergy. This study demonstrates that anti-dsDNA B cells remain tolerant in LT-alpha(-/-), TNF-alpha( /-), and TNFRp55(-/-) mice; however, TNF-alpha or a TNF-alpha-dependent factor is required for their characteristic positioning to the T/B interface. Providing a TNF-alpha signal in TNF-alpha(-/-) mice by systemic administration of an agonist anti-TNFRp55 mAb induces the maturation of the anti-dsDNA B cells and their movement away from the T cell area toward the B cell area. Additionally, the agonist Ab induces changes in the follicular environment, including FDC clustering, up-regulation of the CXC chemokine ligand CXCL13, and down-regulation of the CC chemokine ligands CCL19 and CCL21. Therefore, this study suggests that a balance between B and T cell tropic chemokine signals may be an important mechanism for positioning anergic B cells at the T/B interface of the splenic white pulp. PMID- 11489972 TI - Differential roles of cytokine receptors in the development of epidermal gamma delta T cells. AB - IL-7 and IL-15 play important roles in gammadelta T cell development. These receptors transmit proliferation and/or survival signals in gammadelta T cells. In addition, the IL-7R promotes recombination and transcription in the TCR gamma locus. To clarify the role of the cytokine receptors in the development of epidermal gammadelta T cells, we introduced a Vgamma3/Vdelta1 TCR transgene, derived from Thy-1+ dendritic epidermal T cells (DETC), into IL-7Ralpha-deficient mice, and we found that they partly rescued gammadelta T cells in the adult thymus but not in the spleen. Introduction of an additional Bcl-2 transgene had a minimal effect on gammadelta T cells in the adult thymus of these mice. In contrast to the adult thymus, the introduction of the Vgamma3/Vdelta1 TCR transgene into IL-7Ralpha-/- mice completely restored Vgamma3+ T cells in the fetal thymus and DETC in the adult skin. On the contrary, the same Vgamma3/Vdelta1 TCR transgene failed to rescue DETC in the skin of IL-2Rbeta deficient mice, even with the additional Bcl-2 transgene. These results suggest that the IL-2/IL-15R, rather than the IL-7R, plays an essential role in proliferation and survival of DETC in the fetal thymus and the skin. In contrast, the IL-7R is probably essential in the induction of V-J recombination of TCRgamma genes. Thus, this study proves that IL-7R and IL-2/IL-15R serve differential functions in epidermal gammadelta T cell development. PMID- 11489973 TI - Somatic hypermutation and selection of B cells in thymic germinal centers responding to acetylcholine receptor in myasthenia gravis. AB - The muscle weakness in myasthenia gravis (MG) is mediated by autoantibodies against the nicotinic acetylcholine receptor (AChR) at the neuromuscular junction. Production of these pathogenic autoantibodies is believed to be associated with germinal centers (GC) and anti-AChR-secreting plasma cells in the hyperplastic thymus of patients with early onset MG (EOMG). Here, we describe the repertoire of rearranged heavy chain V genes and their clonal origins in GC from a typical EOMG patient. Three hundred fifteen rearranged Ig V(H) genes were amplified, cloned, and sequenced from sections of four thymic GC containing AChR specific B cells. We found that thymic GC contain a remarkably heterogeneous population of B cells. Both naive and circulating memory B cells undergo Ag driven clonal proliferation, somatic hypermutation, and selection. Numerous B cell clones were present, with no individual clone dominating the response. Comparisons of B cell clonal sequences from different GC and known anti-AChR Abs from other patients showed convergent mutations in the complementarity determining regions. These results are consistent with AChR driving an ongoing GC response in the thymus of EOMG patients. This is the first detailed analysis of B cell clones in human GC responding to a defined protein Ag, and the response we observed may reflect the effects of chronic stimulation by autoantigen. PMID- 11489974 TI - Regulatory T cells induced by 1 alpha,25-dihydroxyvitamin D3 and mycophenolate mofetil treatment mediate transplantation tolerance. AB - 1alpha,25-dihydroxyvitamin D3, the active form of vitamin D3, and mycophenolate mofetil, a selective inhibitor of T and B cell proliferation, modulate APC function and induce dendritic cells (DCs) with a tolerogenic phenotype. Here we show that a short treatment with these agents induces tolerance to fully mismatched mouse islet allografts that is stable to challenge with donor-type spleen cells and allows acceptance of donor-type vascularized heart grafts. Peritransplant macrophages and DCs from tolerant mice express down-regulated CD40, CD80, and CD86 costimulatory molecules. In addition, DCs from the graft area of tolerant mice secrete, upon stimulation with CD4+ cells, 10-fold lower levels of IL-12 compared with DCs from acutely rejecting mice, and induce a CD4+ T cell response characterized by selective abrogation of IFN-gamma production. CD4+ but not CD8+ or class II+ cells from tolerant mice, transferred into naive syngeneic recipients, prevent rejection of donor-type islet grafts. Graft acceptance is associated with impaired development of IFN-gamma-producing type 1 CD4+ and CD8+ cells and an increased percentage of CD4+CD25+ regulatory cells expressing CD152 in the spleen and in the transplant-draining lymph node. Transfer of CD4+CD25+ cells from tolerant but not naive mice protects 100% of the syngeneic recipients from islet allograft rejection. These results demonstrate that a short treatment with immunosuppressive agents, such as 1alpha,25 dihydroxyvitamin D3/mycophenolate mofetil, induces tolerance to islet allografts associated with an increased frequency of CD4+CD25+ regulatory cells that can adoptively transfer transplantation tolerance. PMID- 11489975 TI - Development of the thymus requires signaling through the fibroblast growth factor receptor R2-IIIb. AB - Mice deficient for fibroblast growth factor (Fgf)R2-IIIb show a block in thymic growth after embryonic day 12.5, a stage that just precedes its detection in thymic epithelial cells. Fgf7 and Fgf10, the main ligands for FgfR2-IIIb, are expressed in the mesenchyme surrounding the thymic epithelial primordium, and Fgf10-deficient mice also exhibit impaired thymic growth. Hence, Fgf signaling is essential for thymic epithelial proliferation. In addition to the proliferative block, most thymic epithelial cells fail to progress from an immature cytokeratin 5-positive to a cytokeratin 5-negative phenotype. Nevertheless, sufficient epithelial cell differentiation occurs in the severely hypoplastic thymus to allow the development of CD4/CD8-double-positive thymocytes and a very small number of single-positive thymocytes expressing TCRs. PMID- 11489976 TI - Impairment in the expression and activity of Fyn during differentiation of naive CD4+ T cells into the Th2 subset. AB - We previously showed that the amounts of Fyn protein in Th2 clones were approximately one-third to one-fifth of those in Th1 clones. In this study we examined the role of Fyn in the polarization of naive CD4+ T cells toward the Th2 subset using fyn(-/-) mice. The fyn(-/-) naive CD4+ T cells efficiently produced Th2 cytokines and polarized toward the Th2 subset even in the absence of IL-4 and IL-13. The expression of Fyn in wild-type CD4+ T cells decreased at a transcription level concomitant with polarization toward the Th2 subset. These results suggest that Fyn plays a role in the down-regulation of the differentiation of naive CD4+ T cells into the Th2 subset. PMID- 11489977 TI - Human recent thymic emigrants--identification, expansion, and survival characteristics. AB - This study shows that, in humans at birth, circulating T cells represent recent thymic emigrants (RTEs) as reflected in their high level of expression of TCR excision circles. RTEs express "thymocyte-like" characteristics with regard to rapid rate of apoptosis. In the presence of common gamma-chain cytokines, in particular IL-7, they show enhanced potential to survive, entry into cell cycle, and proliferation. Although common gamma-chain cytokines were also potent antiapoptotic stimuli for mature adult-derived naive CD4+CD45RA+ T cells, these cells were refractory to IL-7-induced expansion in vitro. RTEs cultured with IL-7 could not reinduce recombination-activating gene-2 gene expression in vitro. These data suggest that postthymic naive T cells in the periphery during early life are at a unique stage in ontogeny as RTEs, during which they can undergo homeostatic regulation including expansion and survival in an Ag-independent manner while maintaining their preselected TCR repertoire. PMID- 11489978 TI - Impairment of bleomycin-induced lung fibrosis in CD28-deficient mice. AB - Lung fibrosis is an important pulmonary disease with a high mortality rate, but its pathophysiological mechanism has not been fully clarified. Various types of cells have been implicated in the development of lung fibrosis, including T cells. However, the contribution of functional molecules expressed on T cells to the development of lung fibrosis remains largely unknown. In this study, we determined whether costimulation via CD28 on T cells was crucial for the development of lung fibrosis by intratracheally administering bleomycin into CD28 deficient mice. Compared with wild-type mice, the CD28-deficient mice showed markedly impaired lung fibrosis after injection with low doses of bleomycin, as judged by histological changes and hydroxyproline content in the lungs. In addition, bleomycin-induced T cell infiltration into the airways and production of several cytokines and chemokines including IL-5 were also impaired in the CD28 deficient mice. Furthermore, adoptive transfer of CD28-positive T cells from wild type mice recovered the impaired bleomycin-induced lung fibrosis in CD28 deficient mice. These findings suggest that the CD28-mediated T cell costimulation plays a critical role in the development of lung fibrosis, possibly by regulating the production of cytokines and chemokines in the lung. Thus, manipulation of the CD28-mediated costimulation could be a potential therapeutic strategy for the prevention of lung fibrosis. PMID- 11489979 TI - CD8- dendritic cell activation status plays an integral role in influencing Th2 response development. AB - Whether dendritic cells (DC) play a passive or active role in Th2 response induction is poorly understood. In this study, we show that CD8- DC pulsed with Th2-polarizing Ag (soluble egg Ag (SEA)) from Schistosoma mansoni potently stimulate Th2 responses in vivo and in vitro while failing to undergo a conventional maturation process. Thus, in contrast to DC pulsed with the Th1 response inducing Ag Propionebacterium acnes, SEA-exposed DC exhibit a phenotype that is most similar to that of immature DC, failing to up-regulate expression of CD40, CD54, CD80, CD86, or OX40L; producing no detectable IL-4, IL-10, or IL-12; and displaying only a minor increase in MHC class II expression. Importantly, in vitro derived DC exposed to SEA were phenotypically similar to CD8- DC isolated from active S. mansoni infection. By discriminating between different types of pathogen and responding appropriately, CD8- DC play a major role in the decision process to mount either a Th1 or Th2 response. PMID- 11489980 TI - Signaling lymphocytic activation molecule is expressed on mature CD83+ dendritic cells and is up-regulated by IL-1 beta. AB - Signaling lymphocyte activation molecule (SLAM), a 70-kDa costimulatory molecule that mediates CD28-independent proliferation of T cells and IFN-gamma production, has been identified on human T cells, immature thymocytes, and a subset of B cells. We have found that SLAM is expressed on mature but not immature dendritic cells (DC). However, the SLAM-associated protein, is missing in DC. SLAM surface expression is strongly up-regulated by IL-1beta. Addition of IL-1beta to the DC maturation mixture also increases the stimulatory properties of DC. These findings provide a new marker for DC maturation and help to explain two areas of DC biology. First, SLAM is a receptor for the measles virus, previously shown to infect DC. Second, SLAM could possibly contribute to the enhanced immunostimulatory functions of DC that are observed following the addition of IL 1. PMID- 11489981 TI - Inducible costimulator regulates Th2-mediated inflammation, but not Th2 differentiation, in a model of allergic airway disease. AB - A novel costimulatory molecule expressed on activated T cells, inducible costimulator (ICOS), and its ligand, B7-related protein-1 (B7RP-1), were recently identified. ICOS costimulation leads to the induction of Th2 cytokines without augmentation of IL-2 production, suggesting a role for ICOS in Th2 cell differentiation and expansion. In the present study, a soluble form of murine ICOS, ICOS-Ig, was used to block ICOS/B7RP-1 interactions in a Th2 model of allergic airway disease. In this model, mice are sensitized with inactivated Schistosoma mansoni eggs and are subsequently challenged with soluble S. mansoni egg Ag directly in the airways. Treatment of C57BL/6 mice with ICOS-Ig during sensitization and challenge attenuated airway inflammation, as demonstrated by a decrease in cellular infiltration into the lung tissue and airways, as well as by a decrease in local IL-5 production. These inhibitory effects were not due to a lack of T cell priming nor to a defect in Th2 differentiation. In addition, blockade of ICOS/B7RP-1 interactions during ex vivo restimulation of lung Th2 effector cells prevented cytokine production. Thus, blockade of ICOS signaling can significantly reduce airway inflammation without affecting Th2 differentiation in this model of allergic airway disease. PMID- 11489982 TI - The source of early IFN-gamma that plays a role in Th1 priming. AB - When naive CD4 T cells are primed, they rapidly differentiate into polarized Th1 and/or Th2 phenotypes. A major factor in producing such polarization is the early production of cytokines (IL-12 and IFN-gamma in the case of Th1 cells and IL-4 in the case of Th2 cells). One issue that remains unresolved is the source of the early IFN-gamma that synergizes with IL-12 to fully polarize CD4 T cells into Th1 cells. We have examined this question by injecting mice with anti-CD3 and examining cells from normal and various MHC-knockout mice. We found that IFN gamma is induced rapidly in a small subset of CD8 T cells. This subset is absent in mice that lack beta2-microglobulin, but not in K(b)D(b)-double-knockout mice, indicating that these CD8 T cells are dependent on nonclassical MHC class Ib molecules. The early burst of IFN-gamma polarizes CD4 T cells toward Th1 cells, in part by stimulating the release of IL-12 from APC. We also use TAP- and CD1 knockout mice to show that such cells are not CD1-restricted NK T cells, nor are they dependent on TAP-1 transport for surface expression of the relevant MHC class Ib molecule. Therefore, they arise on MHC class Ib molecules that do not depend on TAP-1 transporters. PMID- 11489983 TI - IFN-gamma-dependent and -independent initiation of switch recombination by NK cells. AB - We have examined the effect of IL-2-propagated NK or NK-T cells on each of the steps required for B cell switch recombination leading to IgG2a production. The results indicate that NK cells, on their own and in the absence of IFN-gamma, can induce germline transcription in resting, IgG(-) B lymphocytes from the gamma2a locus as well as mRNA for activation-induced cytidine deaminase (AID) via a process that requires cell-cell interactions. The results also show that, in contrast to induction by T cells, activation by NK cells does not involve CD40 CD40 ligand interactions and does not extend to the induction of Igamma1 transcription. Furthermore, in contrast to stimulation by LPS and IFN-gamma or by T cells, the activation events initiated by NK cells do not result in significant synthesis of functional gamma2a mRNA in resting B lymphocytes even in the presence of IFN-gamma. Thus, induction of germline and AID transcripts are necessary but not sufficient events for functional switching to IgG2a. These experiments, showing that NK cells themselves cannot induce IgG2a production but can polyclonally program B lymphocytes so that they preferentially switch to this isotype may explain how activated NK cells can skew the Ag-specific immune response toward IgG2a. The findings also provide further demonstration of the definitive yet limited extent of how a non-Ag-specific component of the innate system can modulate the direction of the adaptive immune response. PMID- 11489984 TI - EBV-specific CD8+ T cell memory: relationships between epitope specificity, cell phenotype, and immediate effector function. AB - EBV infection in humans induces CD8+ T cell memory to viral epitopes derived from both lytic and latent cycle Ags. We have analyzed the relationship between the phenotype and function of the memory pool of T cells specific for these Ags. Lytic epitope-specific populations were heterogeneous in terms of CD45RO/RA and CD28 expression, whereas latent epitope-specific populations were uniformly CD45RO+ and CD28+, consistent with the higher antigenic challenge from lytic epitopes driving some memory cells toward a CD45RA+, CD28- phenotype. However, both types of memory population showed immediate epitope-specific cytotoxicity and type 1 cytokine production in ex vivo assays. Cytotoxic function was not associated with preactivated T cells, as EBV-specific populations were negative for activation markers such as CD69 or CD38, nor could cytotoxic function be ascribed to CD27- or CD56+ subsets, as such cells were not detected in EBV specific memory. Furthermore, cytotoxicity was not limited to CD45RA+ and/or CD28 fractions, but also was observed in CD45RO+, CD28+ populations in lytic and latent epitope-specific memory. Cytokine (IFN-gamma, TNF-alpha) responses, measured by intracytoplasmic staining after peptide stimulation, also were detectable in CD45RO+ and RA+ subsets as well as CD28+ and CD28- subsets. Of other markers that were heterogeneous in both lytic and latent epitope populations, CCR7 gave the best discrimination of functionality; thus, CCR7+ cells consistently failed to give an IFN-gamma or TNF-alpha response, whereas many CCR7- cells were responsive. Our data are consistent with effector functions having a broad distribution among phenotypically distinct subsets of "effector memory" cells that have lost the CCR7 marker. PMID- 11489985 TI - Adaptive tolerance of CD4+ T cells in vivo: multiple thresholds in response to a constant level of antigen presentation. AB - The in vivo T cell response to persistent Ag contains a hyporesponsive phase following an initial expansion and subsequent partial deletion of the responding cells. The mechanism(s) responsible for this tolerance process is poorly understood. In this study, we describe a new paired transgenic model (TCR and Ag), which within 7-14 days produces 20-40 million hyporesponsive T cells. This state is characterized by an 85-95% reduction in all cytokine production, an impairment of re-expression of CD25 and CD69, and a desensitization of the proliferative response to Ag. TCR levels were normal, and in vivo mixing experiments showed no evidence for active suppression. The hyporesponsiveness partially dissipated without proliferation when the cells were transferred into a non-Ag-bearing host. If the second host expressed Ag, the T cells initially regained responsiveness, but then slowly entered an even deeper state of tolerance characterized by an additional 7- to 10-fold lowering of cytokine production and a greater desensitization of proliferation. Surprisingly, this readaptation took place with the same level of Ag presentation, suggesting that other parameters can influence the tolerance threshold. Both the readjustment in sensitivity and the reversal without Ag convincingly demonstrate for the first time a truly adaptive tolerance process in CD4+ T cells in vivo. PMID- 11489986 TI - Signaling alterations in activation-induced nonresponsive CD8 T cells. AB - Costimulation-dependent production and autocrine use of IL-2 by activated CD8 T cells results in initial clonal expansion, but this is transient. The cells quickly become anergic, unable to produce IL-2 in response to Ag and costimulation, irrespective of the form of costimulation. This activation-induced non-responsiveness (AINR) differs from "classical" anergy in that it results despite the cells receiving both signal 1 and signal 2. AINR cells can still proliferate in response to exogenous IL-2, but can no longer produce it. Other TCR-mediated events including cytolytic function and IFN-gamma production are not affected in the AINR state. To characterize the mechanism(s) responsible for lack of IL-2 production in CD8 T cells in the AINR state, microspheres bearing immobilized anti-TCR Abs or peptide-MHC complexes, B7-1, and ICAM-1 were used to provide well-defined stimuli to the cells. Comparison of normal and AINR cells revealed that in AINR cells extracellular signal-regulated kinase (ERK) is upregulated more transiently, Janus kinase activation is substantially reduced, and activation of p38 is eliminated. PMA and ionomycin restored proliferation and IL-2 production in AINR cells, indicating a signaling defect upstream of Ras and protein kinase C. Inhibitors of ERK (PD98059) and of p38 kinase (SB202190) blocked IL-2 mRNA expression and proliferation of both peptide-MHC/B7-1/ICAM-1 stimulated normal cells and PMA/ionomycin-stimulated AINR cells. Together these results demonstrate that activation of at least ERK and p38 is essential for IL-2 production by CD8 T cells and that up-regulation of these mitogen-activated protein kinases, along with Janus kinase, is defective in AINR cells. PMID- 11489987 TI - Nicotinamide adenine dinucleotide (NAD) and its metabolites inhibit T lymphocyte proliferation: role of cell surface NAD glycohydrolase and pyrophosphatase activities. AB - The presence of NAD-metabolizing enzymes (e.g., ADP-ribosyltransferase (ART)2) on the surface of immune cells suggests a potential immunomodulatory activity for ecto-NAD or its metabolites at sites of inflammation and cell lysis where extracellular levels of NAD may be high. In vitro, NAD inhibits mitogen stimulated rat T cell proliferation. To investigate the mechanism of inhibition, the effects of NAD and its metabolites on T cell proliferation were studied using ART2a+ and ART2b+ rat T cells. NAD and ADP-ribose, but not nicotinamide, inhibited proliferation of mitogen-activated T cells independent of ART2 allele specific expression. Inhibition by P2 purinergic receptor agonists was comparable to that induced by NAD and ADP-ribose; these compounds were more potent than P1 agonists. Analysis of the NAD-metabolizing activity of intact rat T cells demonstrated that ADP-ribose was the predominant metabolite, consistent with the presence of cell surface NAD glycohydrolase (NADase) activities. Treatment of T cells with phosphatidylinositol-specific phospholipase C removed much of the NADase activity, consistent with at least one NADase having a GPI anchor; ART2- T cell subsets contained NADase activity that was not releasable by phosphatidylinositol-specific phospholipase C treatment. Formation of AMP from NAD and ADP-ribose also occurred, a result of cell surface pyrophosphatase activity. Because AMP and its metabolite, adenosine, were less inhibitory to rat T cell proliferation than was NAD or ADP-ribose, pyrophosphatases may serve a regulatory role in modifying the inhibitory effect of ecto-NAD on T cell activation. These data suggest that T cells express multiple NAD and adenine nucleotide-metabolizing activities that together modulate immune function. PMID- 11489988 TI - Testosterone acts directly on CD4+ T lymphocytes to increase IL-10 production. AB - Males are less susceptible than females to experimental autoimmune encephalomyelitis and many other autoimmune diseases. Gender differences in cytokine production have been observed in splenocytes of experimental autoimmune encephalomyelitis mice stimulated with myelin proteins and may underlie gender differences in susceptibility. As these differences should not be limited to responses specific for myelin proteins, gender differences in cytokine production upon stimulation with Ab to CD3 were examined, and the mechanisms were delineated. Splenocytes from male mice stimulated with Ab to CD3 produced more IL 10 and IL-4 and less IL-12 than those from female mice. Furthermore, splenocytes from dihydrotestosterone (DHT)-treated female mice produced more IL-10 and less IL-12 than those from placebo-treated female mice, whereas there was no difference in IL-4. IL-12 knockout mice were then used to determine whether changes in IL-10 production were mediated directly by testosterone vs indirectly by changes in IL-12. The results of these experiments favored the first hypothesis, because DHT treatment of female IL-12 knockout mice increased IL-10 production. To begin to delineate the mechanism by which DHT may be acting, the cellular source of IL-10 was determined. At both the RNA and protein levels, IL 10 was produced primarily by CD4+ T lymphocytes. CD4+ T lymphocytes were then shown to express the androgen receptor, raising the possibility that testosterone acts directly on CD4+ T lymphocytes to increase IL-10 production. In vitro experiments demonstrated increased IL-10 production following treatment of CD4+ T lymphocytes with DHT. Thus, testosterone can act directly via androgen receptors on CD4+ T lymphocytes to increase IL-10 gene expression. PMID- 11489989 TI - A central role for death receptor-mediated apoptosis in the rejection of tumors by NK cells. AB - NK cells provide a line of defense against tumors and virus-infected cells that have lost the expression of one or more MHC class I isoforms. Here, we investigate whether inhibitors of apoptosis can block the rejection of tumors mediated by NK cells, by introducing the long form of Fas-associated death domain like IL-1beta-converting enzyme-associated inhibitory protein (FLIP(L)) and poxvirus cytokine response modifier A (CrmA) into the MHC class I-deficient T lymphoma cell line RMA-S. RMA-S cells do not normally express Fas in vitro, and it was previously postulated that the rejection of these tumors by NK cells is strictly perforin dependent. We show that perforin-deficient NK cells directly mediate Fas up-regulation on RMA-S cells and thereafter kill the cells in a Fas dependent manner, and that RMA-S FLIP(L) and RMA-S CrmA are protected from such killing. When injected in immunocompetent recipients, RMA-S cells up-regulate Fas, rendering in vivo-passed mock-transduced cells sensitive to Fas-mediated apoptosis. Moreover, RMA-S FLIP(L) and RMA-S CrmA cells establish aggressive tumors, in contrast to RMA-S mock cells that are rejected. These results demonstrate that FLIP(L) and CrmA function as tumor progression factors by protecting MHC class I-deficient tumors from rejection mediated by NK cells. Moreover, our data indicate that death receptor-mediated apoptosis has a more prominent role in the clearance of NK-sensitive tumors than previously suggested. PMID- 11489990 TI - Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent inhibition of IL-5 from human T lymphocytes is not mediated by the cAMP-dependent protein kinase A. AB - IL-5 is implicated in the pathogenesis of asthma and is predominantly released from T lymphocytes of the Th2 phenotype. In anti-CD3 plus anti-CD28-stimulated PBMC, albuterol, isoproterenol, rolipram, PGE2, forskolin, cholera toxin, and the cAMP analog, 8-bromoadenosine cAMP (8-Br-cAMP) all inhibited the release of IL-5 and lymphocyte proliferation. Although all of the above compounds share the ability to increase intracellular cAMP levels and activate protein kinase (PK) A, the PKA inhibitor H-89 failed to ablate the inhibition of IL-5 production mediated by 8-Br-cAMP, rolipram, forskolin, or PGE2. Similarly, H-89 had no effect on the cAMP-mediated inhibition of lymphocyte proliferation. Significantly, these observations occurred at a concentration of H-89 (3 microM) that inhibited both PKA activity and CREB phosphorylation in intact cells. Additional studies showed that the PKA inhibitors H-8, 8-(4-chlorophenylthio) adenosine-3',5'-cyclic monophosphorothioate Rp isomer, and a myristolated PKA inhibitor peptide also failed to block the 8-Br-cAMP-mediated inhibition of IL-5 release from PBMC. Likewise, a role for PKG was considered unlikely because both activators and inhibitors of this enzyme had no effect on IL-5 release. Western blotting identified Rap1, a downstream target of the cAMP-binding proteins, exchange protein directly activated by cAMP/cAMP-guanine nucleotide exchange factors 1 and 2, in PBMC. However, Rap1 activation assays revealed that this pathway is also unlikely to be involved in the cAMP-mediated inhibition of IL-5. Taken together, these results indicate that cAMP-elevating agents inhibit IL-5 release from PBMC by a novel cAMP-dependent mechanism that does not involve the activation of PKA. PMID- 11489991 TI - Il-17 mobilizes peripheral blood stem cells with short- and long-term repopulating ability in mice. AB - Autologous and allogeneic bone marrow transplantations have evolved as important cancer therapy modalities. For both indications, peripheral blood has been shown to have distinct advantages over bone marrow as the stem cell source. Cytokine combinations for mobilization have enhanced stem cell yield and accelerated engraftment. However, novel mobilizing agents and strategies are needed to further improve clinical outcomes. Within the donor graft, the dynamic equilibrium between T cells and stem cells critically influences engraftment and transplantation results. IL-17 is a cytokine produced almost exclusively from activated T cells. IL-17 was expressed in vivo with adenovirus technology. Here, proof-of-principle studies demonstrate that IL-17 effectively mobilizes hemopoietic precursor cells (CFU-granulocyte-erythrocyte-macrophage-monocyte, CFU high proliferative potential) and primitive hemopoietic stem cells (Lin(-/low)c kit(+)Sca1(+)). Moreover, mouse IL-17 adenovirus-mobilized peripheral blood stem cells rescued lethally irradiated mice. Bone marrow was found to be 45-75% of donor origin at 1 year. In secondary recipients, donor-derived bone marrow cells ranged from 45 to 95%. These data show that IL-17 mobilizes stem cells in mice with short- and long-term reconstituting capacity. Additional comparative studies are needed as well as studies in tumor models to refine distinct potential clinical applications for IL-17-mobilized peripheral blood stem cells. PMID- 11489992 TI - Predominance of NK1.1+TCR alpha beta+ or DX5+TCR alpha beta+ T cells in mice conditioned with fractionated lymphoid irradiation protects against graft-versus host disease: "natural suppressor" cells. AB - We developed a nonmyeloablative host conditioning regimen in a mouse model of MHC mismatched bone marrow transplantation that not only reduces radiation toxicity, but also protects against graft-vs-host disease. The regimen of fractionated irradiation directed to the lymphoid tissues and depletive anti-T cell Abs results in a marked change in the residual host T cells, such that NK1.1+ or DX5+asialo-GM1+ T cells become the predominant T cell subset in the lymphoid tissues of C57BL/6 and BALB/c mice, respectively. The latter "natural suppressor" T cells protect hosts from graft-vs-host disease after the infusion of allogeneic bone marrow and peripheral blood cells that ordinarily kill hosts conditioned with sublethal or lethal total body irradiation. Protected hosts become stable mixed chimeras, but fail to show the early expansion and infiltration of donor T cells in the gut, liver, and blood associated with host tissue injury. Cytokine secretion and adoptive transfer studies using wild-type and IL-4(-/-) mice showed that protection afforded by NK1.1+ and DX5+asialo-GM1+ T cells derived from either donors or hosts conditioned with lymphoid irradiation is dependent on their secretion of high levels of IL-4. PMID- 11489993 TI - Tapasin enhances peptide-induced expression of H2-M3 molecules, but is not required for the retention of open conformers. AB - H2-M3 is a class Ib MHC molecule that binds a highly restricted pool of peptides, resulting in its intracellular retention under normal conditions. However, addition of exogenous M3 ligands induces its escape from the endoplasmic reticulum (ER) and, ultimately, its expression at the cell surface. These features of M3 make it a powerful and novel model system to study the potentially interrelated functions of the ER-resident class I chaperone tapasin. The functions ascribed to tapasin include: 1) ER retention of peptide-empty class I molecules, 2) TAP stabilization resulting in increased peptide transport, 3) direct facilitation of peptide binding by class I, and 4) peptide editing. We report in this study that M3 is associated with the peptide-loading complex and that incubation of live cells with M3 ligands dramatically decreased this association. Furthermore, high levels of open conformers of M3 were efficiently retained intracellularly in tapasin-deficient cells, and addition of exogenous M3 ligands resulted in substantial surface induction that was enhanced by coexpression of either membrane-bound or soluble tapasin. Thus, in the case of M3, tapasin directly facilitates intracellular peptide binding, but is not required for intracellular retention of open conformers. As an alternative approach to define unique aspects of M3 biosynthesis, M3 was expressed in human cell lines that lack an M3 ortholog, but support expression of murine class Ia molecules. Unexpectedly, peptide-induced surface expression of M3 was observed in only one of two cell lines. These results demonstrate that M3 expression is dependent on a unique factor compared with class Ia molecules. PMID- 11489994 TI - A point mutation in the IL-12R beta 2 gene underlies the IL-12 unresponsiveness of Lps-defective C57BL/10ScCr mice. AB - Lps-defective C57BL/10ScCr (Cr) mice are homozygous for a deletion encompassing Toll-like receptor 4 that makes them refractory to the biological activity of LPS. In addition, these mice exhibit an inherited IL-12 unresponsiveness resulting in impaired IFN-gamma responses to different microorganisms. By positional cloning methods, we show here that this second defect of Cr mice is due to a mutation in a single gene located on mouse chromosome 6, in close proximity to the Igkappa locus. The gene is IL-12Rbeta2. Cr mice carry a point mutation creating a stop codon that is predicted to cause premature termination of the translated IL-12Rbeta2 after a lysine residue at position 777. The truncated beta2 chain can still form a heterodimeric IL-12R that allows phosphorylation of Janus kinase 2, but, unlike the wild-type IL-12R, can no longer mediate phosphorylation of STAT4. Because the phosphorylation of STAT4 is a prerequisite for the IL-12-mediated induction of IFN-gamma, its absence in Cr mice is responsible for their defective IFN-gamma response to microorganisms. PMID- 11489995 TI - Isolation and characterization of a novel IgD-binding protein from Moraxella catarrhalis. AB - A novel surface protein of the bacterial species Moraxella catarrhalis that displays a high affinity for IgD (MID) was solubilized in Empigen and isolated by ion exchange chromatography and gel filtration. The apparent molecular mass of monomeric MID was estimated to approximately 200 kDa by SDS-PAGE. The mid gene was cloned and expressed in Escherichia coli. The complete mid nucleotide gene sequence was determined, and the deduced amino acid sequence consists of 2123 residues. The sequence of MID has no similarity to other Ig-binding proteins and differs from all previously described outer membrane proteins of M. catarrhalis. MID was found to exhibit unique Ig-binding properties. Thus, in ELISA, dot blots, and Western blots, MID bound two purified IgD myeloma proteins, four IgD myeloma sera, and finally one IgD standard serum. No binding of MID was detected to IgG, IgM, IgA, or IgE myeloma proteins. MID also bound to the surface-expressed B cell receptor IgD, but not to other membrane molecules on human PBLs. This novel Ig binding reagent promises to be of theoretical and practical interest in immunological research. PMID- 11489996 TI - Distinct control of the frequency and allelic exclusion of the V beta gene rearrangement at the TCR beta locus. AB - Ag receptor gene loci contain many V gene segments, each of which is recombined and expressed at a different frequency and is subject to allelic exclusion. To probe the parameters that mediate the different levels of regulation of V gene rearrangement, a Vbeta gene segment together with 3.6-kb 5' and 0.7-kb 3' flanking sequences was inserted 6.8 kb upstream of the Dbeta1 gene segment in the murine TCRbeta locus. Despite its proximity to the Dbeta gene segments and the Ebeta enhancer, the inserted Vbeta segment underwent VDJ recombination at the same frequency as the natural copy located 470 kb upstream. However, the inserted Vbeta segment was no longer under allelic exclusion control as it recombined at a similar frequency in the presence of a TCRbeta transgene. These results suggest that while the inserted fragment contains the necessary cis-regulatory elements for determining the frequency of Vbeta rearrangement, additional cis-regulatory elements are required for mediating Vbeta allelic exclusion. Interestingly, most of the inserted Vbeta rearrangements were not transcribed and expressed in the presence of a TCRbeta transgene, suggesting that TCRbeta allelic exclusion can also be achieved by blocking the transcription of the rearranged gene segments. These findings provide strong evidence for distinct control of the frequency and allelic exclusion of Vbeta gene rearrangement. PMID- 11489997 TI - Combinatorial peptide libraries and biometric score matrices permit the quantitative analysis of specific and degenerate interactions between clonotypic TCR and MHC peptide ligands. AB - The interaction of TCRs with MHC peptide ligands can be highly flexible, so that many different peptides are recognized by the same TCR in the context of a single restriction element. We provide a quantitative description of such interactions, which allows the identification of T cell epitopes and molecular mimics. The response of T cell clones to positional scanning synthetic combinatorial libraries is analyzed with a mathematical approach that is based on a model of independent contribution of individual amino acids to peptide Ag recognition. This biometric analysis compares the information derived from these libraries composed of trillions of decapeptides with all the millions of decapeptides contained in a protein database to rank and predict the most stimulatory peptides for a given T cell clone. We demonstrate the predictive power of the novel strategy and show that, together with gene expression profiling by cDNA microarrays, it leads to the identification of novel candidate autoantigens in the inflammatory autoimmune disease, multiple sclerosis. PMID- 11489998 TI - SCID in Jack Russell terriers: a new animal model of DNA-PKcs deficiency. AB - We recently described the incidence of a SCID disease in a litter of Jack Russell terriers. In this study, we show that the molecular defect in these animals is faulty V(D)J recombination. Furthermore, we document a complete deficit in DNA dependent protein kinase activity that can be explained by a marked diminution in the expression of the catalytic subunit DNA-dependent protein kinase catalytic subunit (DNA-PKcs). We conclude that as is the case in C.B-17 SCID mice and in Arabian SCID foals, the defective factor in these SCID puppies is DNA-PKcs. In mice, it has been clearly established that DNA-PKcs deficiency produces an incomplete block in V(D)J recombination, resulting in "leaky" coding joint formation and only a modest defect in signal end ligation. In contrast, DNA-PKcs deficiency in horses profoundly blocks both coding and signal end joining. Here, we show that although DNA-PKcs deficiency in canine lymphocytes results in a block in both coding and signal end joining, the deficit in both is intermediate between that seen in SCID mice and SCID foals. These data demonstrate significant species variation in the absolute necessity for DNA-PKcs during V(D)J recombination. Furthermore, the severity of the V(D)J recombination deficits in these three examples of genetic DNA-PKcs deficiency inversely correlates with the relative DNA-PK enzymatic activity expressed in normal fibroblasts derived from these three species. PMID- 11489999 TI - Circulating human B cells that express surrogate light chains display a unique antibody repertoire. AB - Circulating human B cells that coexpress V-preB and conventional L chains (V preB+L+ B cells) are a recently described subset of B cells that express Abs with features of self-reactivity. Initial analysis of V-preB+L+ B cells was limited to Ig-kappa and to the small, underused VH5 family. To determine whether Abs commonly expressed by V-preB+L+ B cells show similar features, we analyzed Ig H chains from three highly expressed VH families, VH1, VH3, and VH4, and Ig-lambda. We find that VH1 and VH3 Abs expressed by V-preB+L+ B cells resemble VH5 in that they display increased JH6 use, long CDR3s, and an increased frequency of D-D fusions. Abs in all three of these VH families also show skewed D reading frame use resulting in predominance of hydrophobic amino acids, which are counterselected in conventional B cells. Like Ig-kappa genes, the Ig-lambda genes in V-preB+L+ B cells show long CDR3s, but they differ from Ig-kappa genes in that they display no evidence of receptor editing. We conclude that a large number of H and L chain Abs expressed by V-preB+L+ B cells display features associated with self-reactive Abs. PMID- 11490000 TI - E2A and HEB activate the pre-TCR alpha promoter during immature T cell development. AB - The pre-TCRalpha (pTalpha) is exclusively expressed in immature thymocytes and constitutes the pre-TCR complex with TCRbeta, which regulates early T cell differentiation. Despite the recent identification of the pTalpha enhancer, the contribution of the promoter region, the direct DNA-protein interaction, and the regulation of such interaction along with T cell development have not been investigated. We analyzed the pTalpha promoter region and identified the critical elements for transcription of the pTalpha gene. The pTalpha promoter was found to contain two consecutive E-box elements that are critical for pTalpha transcription. The E-box elements in the promoter region formed the specific DNA protein complex that was exclusively observed in immature thymocytes, not in mature thymocytes and T cells. The E proteins in this complex were identified as E2A and HeLa E-box binding protein (HEB), and overexpression of E2A and HEB resulted in activation of the pTalpha promoter. The binding complex in the consecutive E-boxes in the pTalpha promoter changed along with T cell development, as a distinct DNA-binding complex was observed in mature T cells. Comparing the E-box regions in the enhancer and the promoter, those in the promoter appear to make a greater contribution to pTalpha gene transcription. PMID- 11490001 TI - Characterization of the active sites in decay-accelerating factor. AB - Decay-accelerating factor (DAF) is a complement regulator that dissociates autologous C3 convertases, which assemble on self cell surfaces. Its activity resides in the last three of its four complement control protein repeats (CCP2 4). Previous modeling on the nuclear magnetic resonance structure of CCP15-16 in the serum C3 convertase regulator factor H proposed a positively charged surface area on CCP2 extending into CCP3, and hydrophobic moieties between CCPs 2 and 3 as being primary convertase-interactive sites. To map the residues providing for the activity of DAF, we analyzed the functions of 31 primarily alanine substitution mutants based in part on this model. Replacing R69, R96, R100, and K127 in the positively charged CCP2-3 groove or hydrophobic F148 and L171 in CCP3 markedly impaired the function of DAF in both activation pathways. Significantly, mutations of K126 and F169 and of R206 and R212 in downstream CCP4 selectively reduced alternative pathway activity without affecting classical pathway activity. Rhesus macaque DAF has all the above human critical residues except for F169, which is an L, and its CCPs exhibited full activity against the human classical pathway C3 convertase. The recombinants whose function was preferentially impaired against the alternative pathway C3bBb compared with the classical pathway C4b2a were tested in classical pathway C5 convertase (C4b2a3b) assays. The effects on C4b2a and C4b2a3b were comparable, indicating that DAF functions similarly on the two enzymes. When CCP2-3 of DAF were oriented according to the crystal structure of CCP1-2 of membrane cofactor protein, the essential residues formed a contiguous region, suggesting a similar spatial relationship. PMID- 11490002 TI - Human cathepsin W, a cysteine protease predominantly expressed in NK cells, is mainly localized in the endoplasmic reticulum. AB - Human cathepsin W (also called lymphopain) is a recently described papain-like cysteine protease of unknown function whose gene expression was found to be restricted to cytotoxic cells. Here we demonstrate that cathepsin W is expressed predominantly in NK cells and, to a lesser extent, in CTLs. Quantitative RT-PCR revealed that NK cells contained approximately 21 times more cathepsin W transcript than CTLs. The predominant expression of cathepsin W in NK cells was further confirmed by Western blot analysis and immunohistochemistry. IL-2 mediated stimulation of NK cells and CTLs revealed a stronger up-regulation of the cathepsin W gene and protein expression in NK cells (7-fold) than in CTLs (2 fold). Transfection experiments of HeLa cells and biochemical analyses revealed that cathepsin W is exclusively "high mannose-type" glycosylated and is mainly targeted to the endoplasmic reticulum (ER). Interestingly, the ER localization of cathepsin W was also found in NK cells, in which colocalization studies revealed an overlapping staining of cathepsin W and Con A, an ER-specific lectin. Furthermore, subcellular fractionation of cathepsin W-expressing cells confirmed the ER localization and showed that cathepsin W is membrane associated. Based on the results of this study, cathepsin W might represent a putative component of the ER-resident proteolytic machinery. The constitutive expression in NK cells and the stronger up-regulation of cathepsin W by IL-2 in NK cells than CTLs suggest that cathepsin W is not just a marker of cytotoxic cells but is, rather, specifically expressed in NK cells. PMID- 11490003 TI - Mutation of a single conserved residue in VH complementarity-determining region 2 results in a severe Ig secretion defect. AB - During an immune response, somatic mutations are introduced into the VH and VL regions of Ig chains. The consequences of somatic mutation in highly conserved residues are poorly understood. Ile51 is present in 91% of murine VH complementarity-determining region 2 sequences, and we demonstrate that single Ile51-->Arg or Lys substitutions in the PCG1-1 Ab are sufficient to severely reduce Ig secretion (1-3% of wild-type (WT) levels). Mutant H chains, expressed in the presence of excess L chain, associate with Ig binding protein (BiP) and GRP94 and fail to form HL and H2L assembly intermediates efficiently. The mutations do not irreversibly alter the VH domain as the small amount of mutant H chain, which assembles with L chain as H2L2, is secreted. The secreted mutant Ab binds phosphocholine-protein with avidity identical with that of WT Ab, suggesting that the combining site adopts a WT conformation. A computer-generated model of the PCG1-1 variable region fragment of Ig (Fv) indicates that Ile51 is buried between complementarity-determining region 2 and framework 3 and does not directly contact the L chain. Thus, the Ile51-->Arg or Ile51-->Lys mutations impair association with the PCG1-1 L chain via indirect interactions. These interactions are in part dependent on the nature of the L chain as the PCG1-1 VH single Ile51-->Arg or Ile51-->Lys mutants were partially rescued when expressed with the J558L lambda1 L chain. These results represent the first demonstration that single somatic mutations in V(H) residues can impair Ig secretion and suggest one reason for the conservation of Ile51 in so many Ig VH. PMID- 11490004 TI - Mycobacterial lysocardiolipin is exported from phagosomes upon cleavage of cardiolipin by a macrophage-derived lysosomal phospholipase A2. AB - Pathogenic mycobacteria are able to survive and proliferate in phagosomes within host macrophages (Mphi). This capability has been attributed in part to their cell wall, which consists of various unique lipids. Some of these are important in the host-pathogen interaction, such as resistance against microbicidal effector mechanisms and modulation of host cell functions, and/or are presented as Ags to T cells. Here we show that two lipids are released from the mycobacterial cell wall within the phagosome of infected Mphi and transported out of this compartment into intracellular vesicles. One of these lipids was identified as lysocardiolipin. Lysocardiolipin was generated through cleavage of mycobacterial cardiolipin by a Ca2+-independent phospholipase A2 present in Mphi lysosomes. This result indicates that lysosomal host cell enzymes can interact with released mycobacterial lipids to generate new products with a different intracellular distribution. This represents a novel pathway for the modification of bacterial lipid Ags. PMID- 11490005 TI - Toxoplasma gondii tachyzoites inhibit proinflammatory cytokine induction in infected macrophages by preventing nuclear translocation of the transcription factor NF-kappa B. AB - Control of microbial infection requires regulated induction of NF-kappaB dependent proinflammatory cytokines such as IL-12 and TNF-alpha. Activation of this important transcription factor is driven by phosphorylation-dependent degradation of the inhibitory IkappaB molecule, an event which enables NF-kappaB translocation from the cytoplasm to the nucleus. In this study, we show that intracellular infection of macrophages with the protozoan parasite Toxoplasma gondii induces rapid IkappaB phosphorylation and degradation. Nevertheless, NF kappaB failed to translocate to the nucleus, enabling the parasite to invade cells without triggering proinflammatory cytokine induction. Infected cells subsequently subjected to LPS triggering were severely crippled in IL-12 and TNF alpha production, a result of tachyzoite-induced blockade of NF-kappaB nuclear translocation. Our results are the first to demonstrate the ability of an intracellular protozoan to actively interfere with the NF-kappaB activation pathway in macrophages, an activity that may enable parasite survival within the host. PMID- 11490006 TI - Expression of TNF-alpha by herpes simplex virus-infected macrophages is regulated by a dual mechanism: transcriptional regulation by NF-kappa B and activating transcription factor 2/Jun and translational regulation through the AU-rich region of the 3' untranslated region. AB - Here we have investigated the regulation of TNF-alpha expression in macrophages during HSV-2 infection. Despite a low basal level of TNF-alpha mRNA present in resting macrophages, no TNF-alpha protein is detectable. HSV-2 infection marginally increases the level of TNF-alpha mRNA and protein in resting macrophages, whereas a strong increase is observed in IFN-gamma-activated cells infected with the virus. By reporter gene assay it was found that HSV infection augments TNF-alpha promoter activity. Moreover, treatment of the cells with actinomycin D, which totally blocked mRNA synthesis, only partially prevented accumulation of TNF-alpha protein, indicating that the infection lifts a block on translation of TNF-alpha mRNA. EMSA analysis showed that specific binding to the kappaB#3 site of the murine TNF-alpha promoter was induced within 1 h after infection and persisted beyond 5 h where TNF-alpha expression is down-modulated. Binding to the cAMP responsive element site was also induced but more transiently with kinetics closely following activation of the TNF-alpha promoter. Inhibitors against either NF-kappaB activation or the activating transcription factor 2 kinase p38 abrogated TNF-alpha expression, showing a requirement for both signals for activation of the promoter. This observation was corroborated by reporter gene assays. As to the translational regulation of TNF-alpha, the AU-rich sequence in the 3' untranslated region of the mRNA was found to be responsible for this control because deletion of this region renders mRNA constitutively translationable. These results show that TNF-alpha production is induced by HSV-2 in macrophages through both transcriptional and translational regulation. PMID- 11490007 TI - Dynamic antigen presentation patterns of Listeria monocytogenes-derived CD8 T cell epitopes in vivo. AB - Little information exists regarding the presentation of antigenic peptides in infected tissues. In this study the in vivo presentation of four different CD8 T cell epitopes of Listeria monocytogenes was monitored. Peptide presentation was measured by a new, highly sensitive, ex vivo Ag presentation assay that was based on the testing of freshly isolated cells from infected spleens with peptide specific CD8 T cell lines in an IFN-gamma-specific ELISPOT assay. Remarkably, the peptide presentation pattern of splenocytes and that of macrophages purified from spleens of L. monocytogenes-infected mice were different from those of in vitro infected macrophage-like cell lines. The in vivo Ag presentation pattern of splenocytes also exhibited dynamic changes during the first 48 h of infection. In vivo peptide presentation at later time points postinfection was biased toward immunodominant CD8 T cell epitopes, while at an early time point, 6 h postinfection, subdominant and dominant CD8 T cell epitopes were presented with similar strength. In summary, our studies show that Ag presentation during an infection is a highly dynamic process that only can be fully appreciated by the study of cells infected in their physiological environment. PMID- 11490008 TI - V beta 6+ T cells are obligatory for vaccine-induced immunity to Histoplasma capsulatum. AB - We examined TCR usage to a protective fragment of heat shock protein 60 from the fungus, Histoplasma capsulatum. Nearly 90% of T cell clones from C57BL/6 mice vaccinated with this protein were Vbeta6+; the remainder were Vbeta14+. Amino acid motifs of the CDR3 region from Vbeta6+ cells were predominantly IxGGG, IGG, or SxxGG, whereas it was uniformly SFSGG for Vbeta14+ clones. Short term T cell lines from Vbeta6+-depleted mice failed to recognize Ag, and no T cell clones could be generated. To determine whether Vbeta6+ cells were functionally important, we eliminated them during vaccination. Depletion of Vbeta6+ cells abrogated protection in vivo and upon adoptive transfer of cells into TCR alphabeta(-/-) mice. Transfer of a Vbeta6+, but not a Vbeta14+, clone into TCR alphabeta(-/-) mice prolonged survival. Cytokine generation by Ag-stimulated splenocytes from immunized mice depleted of Vbeta6+ cells was similar to that of controls. The efficacy of the Vbeta6+ clone was associated with elevated production of IFN-gamma, TNF-alpha, and GM-CSF compared with that of the Vbeta14+ clone. More Vbeta6+ cells were present in lungs and spleens of TCR alphabeta(-/-) on day 3 postinfection compared with Vbeta14+ cells. Thus, a single Vbeta family was essential for vaccine-induced immunity. Moreover, the mechanism by which Vbeta6+ contributed to protective immunity differed between unfractionated splenocytes and T cell clones. PMID- 11490009 TI - Functional transitions in macrophages during in vivo infection with Mycobacterium bovis bacillus Calmette-Guerin. AB - Macrophage activation during the immune response to intracellular bacteria is critical for resolution of the infection. We have investigated the pathway of macrophage activation during murine Mycobacterium bovis bacillus Calmette-Guerin (BCG) infection. Three distinct phenotypes of macrophages were identified and compared: resident peritoneal macrophages, day 2 postinfection macrophages, and 12-day postinfection macrophages. Compared with resident peritoneal macrophages, day 2 BCG macrophages expressed intermediate levels of the cell surface receptors Mac1 and F4/80 and low levels of MHC class II molecules. These cells were highly phagocytic and produced large amounts of mRNA encoding the chemokine IP-10. In addition, day 2 BCG macrophages did not generate reactive nitrogen intermediates, though they were primed to do so, and did not have increased levels of TNF-alpha mRNA. Blockade of monocyte influx into the peritoneal cavity using Abs to platelet endothelial cell adhesion molecule 1 had no effect on the appearance of day 2 BCG macrophages, suggesting this cell can differentiate from resident peritoneal macrophages. In contrast to day 2 BCG macrophages, day 12 BCG macrophages were poorly phagocytic, but produced high levels of reactive nitrogen intermediates, IP-10 and TNF-alpha mRNA, and class II MHC molecules. We propose that day 2 BCG macrophages are specialized for phagocytic uptake of pathogens from the extracellular space, whereas day 12 BCG macrophages are specialized for killing of the internalized pathogens. This functional transition during activation is reminiscent of that seen during maturation/activation of the related dendritic cell lineage induced by bacterial or inflammatory stimuli. PMID- 11490010 TI - The role of IL-4 in Heligmosomoides polygyrus-induced alterations in murine intestinal epithelial cell function. AB - IL-4 and IL-13 promote gastrointestinal worm expulsion, at least in part, through effects on nonlymphoid cells, such as intestinal epithelial cells. The role of IL 4/IL-13 in the regulation of intestinal epithelial function during Heligmosomoides polygyrus (Hp) infection was investigated in BALB/c mice infected with Hp or treated with a long-lasting formulation of recombinant mouse IL 4/alphaIL-4 complexes (IL-4C) for 7 days. Separate groups of BALB/c mice were drug-cured of initial infection and later reinfected and treated with anti-IL-4R mAb, an antagonist of IL-4 and IL-13 receptor binding, or with a control mAb. Segments of jejunum were mounted in Ussing chambers, and short circuit current responses to acetylcholine, histamine, serotonin, PGE2, and glucose were determined. Although only modest changes in epithelial cell function were observed during primary Hp infection, IL-4C or a secondary Hp infection each induced more dramatic changes, including increased mucosal permeability, reduced sodium-linked glucose absorption, and increased Cl- secretory response to PGE2. Some, but not all, effects of IL-4C and Hp infection were dependent on enteric nerves. Hp-induced changes in epithelial function were attenuated or prevented by anti-IL-4R mAb. Thus, IL-4/IL-13 mediate many of the effects of Hp infection on intestinal epithelial cell function and do so both through direct effects on epithelial cells and through indirect, enteric nerve-mediated prosecretory effects. These immune system-independent effector functions of IL-4/IL-13 may be important for host protection against gastrointestinal nematodes. PMID- 11490011 TI - Targeting of Pseudomonas aeruginosa in the bloodstream with bispecific monoclonal antibodies. AB - We examined the ability of a bispecific mAb reagent, consisting of a mAb specific for the primate erythrocyte complement receptor cross-linked with an anti bacterial mAb, to target bacteria in the bloodstream in an acute infusion model in monkeys. In vitro studies demonstrated a variable level of complement-mediated binding (immune adherence) of Pseudomonas aeruginosa (strain PAO1) to primate E in serum. In vivo experiments in animals depleted of complement revealed that binding of bacteria to E was <1% before administration of the bispecific reagent, but within 5 min of its infusion, >99% of the bacteria bound to E. In complement replete monkeys, a variable fraction of infused bacteria bound to E. This finding may have significant implications in the interpretation of animal models and in the understanding of bacteremias in humans. Treatment of these complement-replete monkeys with the bispecific reagent led to >99% binding of bacteria to E. Twenty four-hour survival studies were conducted; several clinical parameters, including the degree of lung damage, cytokine levels, and liver enzymes in the circulation, indicate that the bispecific mAb reagent provides a degree of protection against the bacterial challenge. PMID- 11490012 TI - Protective roles of mast cells against enterobacterial infection are mediated by Toll-like receptor 4. AB - Toll-like receptors (TLRs) are mammalian homologues of the Drosophila Toll receptors and are thought to have roles in innate recognition of bacteria. We demonstrated that TLR 2, 4, 6, and 8 but not TLR5 were expressed on mouse bone marrow-derived mast cells (BMMCs). Using BMMCs from the genetically TLR4-mutated strain C3H/HeJ, we demonstrated that functional TLR4 was required for a full responsiveness of BMMCs to produce inflammatory cytokines (IL-1beta, TNF-alpha, IL-6, and IL-13) by LPS stimulation. TLR4-mediated stimulation of mast cells by LPS was followed by activation of NF-kappaB but not by stress-activated protein kinase/c-Jun NH2-terminal kinase signaling. In addition, in the cecal ligation and puncture-induced acute septic peritonitis model, we demonstrated that genetically mast cell-deficient W/W(v) mice that were reconstituted with TLR4 mutated BMMCs had significantly higher mortality than W/W(v) mice reconstituted with TLR4-intact BMMCs. Higher mortality of TLR4-mutated BMMC-reconstituted W/W(v) mice was well correlated with defective neutrophil recruitment and production of proinflammatory cytokines in the peritoneal cavity. Taken together, these observations provide definitive evidence that mast cells play important roles in exerting the innate immunity by releasing inflammatory cytokines and recruitment of neutrophils after recognition of enterobacteria through TLR4 on mast cells. PMID- 11490013 TI - Induction of tolerance to lipopolysaccharide and mycobacterial components in Chinese hamster ovary/CD14 cells is not affected by overexpression of Toll-like receptors 2 or 4. AB - Down-regulation of cell surface expression of Toll-like receptor (TLR) 4 following LPS stimulation has been suggested to underlie endotoxin tolerance. In this study, we examined whether overexpression of TLR2 or TLR4 would affect the ability of cells to become tolerant to LPS or the mycobacterial components, arabinose-capped lipoarabinomannan (LAM) and soluble tuberculosis factor (STF). To this end, Chinese hamster ovary/CD14 cells stably transfected with a NF-kappaB dependent reporter construct, endothelial leukocyte adhesion molecule CD25 (the 3E10 clone), were engineered to overexpress either human TLR2 or TLR4. Transfected TLRs exhibited proper signaling functions, as evidenced by increased LPS responsiveness of 3E10/TLR4 cells and acquisition of sensitivity to TLR2 specific ligands upon transfection of TLR2 into TLR2-negative 3E10 cells. Pretreatment of cells with LPS, LAM, or STF did not modulate TLR2 or TLR4 cell surface expression. Following LPS exposure, 3E10, 3E10/TLR2, and 3E10/TLR4 cells exhibited comparable decreases in LPS-mediated NF-kappaB activation and mitogen activated protein (MAP) kinase phosphorylation. Likewise, LPS pretreatment profoundly inhibited LPS-induced NF-kappaB translocation in Chinese hamster ovary cells that concomitantly overexpressed human TLR4 and myeloid differentiation protein-2 (MD-2), but failed to modulate TLR4 or MD-2 cell surface expression. Pretreatment of 3E10/TLR2 cells with LAM or STF decreased their NF-kappaB responses induced by subsequent stimulation with these substances or LPS. Conversely, prior exposure of 3E10/TLR2 cells to LPS led to hyporesponsiveness to LPS, LAM, and STF, indicating that LPS and mycobacterial products induce cross tolerance. Thus, tolerance to LPS and mycobacterial components cannot be attributed solely to a decrease in TLR/MD-2 expression levels, suggesting inhibition of expression or function of other signaling intermediates. PMID- 11490014 TI - Differential requirements for core2 glucosaminyltransferase for endothelial L selectin ligand function in vivo. AB - L-selectin is a calcium-dependent lectin on leukocytes mediating leukocyte rolling in high endothelial venules and inflamed microvessels. Many selectin ligands require modification of glycoproteins by leukocyte core2 beta1,6-N acetylglucosaminyltransferase (Core2GlcNAcT-I). To test the role of Core2GlcNAcT I for L-selectin ligand biosynthesis, we investigated leukocyte rolling in venules of untreated and TNF-alpha-treated cremaster muscles and in Peyer's patch high endothelial venules (HEV) of Core2GlcNAcT-I null (core2(-/-)) mice. In the presence of blocking mAbs against P- and E-selectin, L-selectin-mediated leukocyte rolling was almost completely abolished in cremaster muscle venules of core2(-/-) mice, but not littermate control mice. By contrast, leukocyte rolling in Peyer's patch HEV was not significantly different between core2(-/-) and control mice. To probe L-selectin ligands more directly, we injected L-selectin coated beads. These beads showed no rolling in cremaster muscle venules of core2( /-) mice, but significant rolling in controls. In Peyer's patch HEV, beads coated with a low concentration of L-selectin showed reduced rolling in core2(-/-) mice. Beads coated with a 10-fold higher concentration of L-selectin rolled equivalently in core2(-/-) and control mice. Our data show that endothelial L selectin ligands relevant for rolling in inflamed microvessels of the cremaster muscle are completely Core2GlcNAcT-I dependent. In contrast, L-selectin ligands in Peyer's patch HEV are only marginally affected by the absence of Core2GlcNAcT I, but are sufficiently functional to support L-selectin-dependent leukocyte rolling in Core2GlcNAcT-I-deficient mice. PMID- 11490015 TI - Taurine chloramine inhibits inducible nitric oxide synthase and TNF-alpha gene expression in activated alveolar macrophages: decreased NF-kappaB activation and IkappaB kinase activity. AB - Taurine prevents tissue damage in a variety of models that involve inflammation, including oxidant-induced lung damage. The mechanism of protection is uncertain, but is postulated to involve the actions of taurine chloramine (Tau-Cl) derived via halide-dependent myeloperoxidase associated with neutrophils. Understanding the influence of Tau-Cl on the production of inflammatory mediators by alveolar macrophages provides an opportunity for determining the mechanism of Tau-Cl action. The effects of Tau-Cl were evaluated on the production of NO and TNF alpha in NR8383, a cloned cell line derived from rat alveolar macrophages (RAM), and in primary cultures of RAM. Production of NO and TNF-alpha, and expression of inducible NO synthase was inhibited by Tau-Cl in activated NR8383 cells as well as in RAM. Temporal (2, 4, 8, 24 h) expression of inducible NO synthase and TNF alpha mRNAs was reduced by Tau-Cl in NR8383 cells. Tau-Cl depressed NF-kappaB migration into the nucleus of activated NR8383 cells and caused a more sustained presence of IkappaB in the cytoplasm. Stabilization of cytoplasmic IkappaB-alpha in Tau-Cl-treated cells resulted from decreased phosphorylation of IkappaB-alpha serine-32 and a lower activity of IkappaB kinase (IKK). Additional experiments demonstrated that Tau-Cl does not directly inhibit IKK activity. These results suggest that Tau-Cl exerts its effects at some level upstream of IKK in the signaling pathway and inhibits production of inflammatory mediators through a mechanism that, at least in part, involves inhibition of NF-kappaB activation. PMID- 11490016 TI - Differences in functional consequences and signal transduction induced by IL-3, IL-5, and nerve growth factor in human basophils. AB - Previous studies have indicated a redundancy in the effects of the cytokines, IL 3, IL-5, and nerve growth factor (NGF) on acute priming of human basophils. In the current study, we have examined the effects of these three cytokines on 18-h priming for leukotriene C4 generation, their ability to induce Fc(epsilon)RIbeta mRNA expression, or their ability to sustain basophil viability in culture. We also examine a variety of the signaling steps that accompany activation with these cytokines. In contrast with the ability of IL-3 to alter secretagogue mediated cytosolic calcium responses following 18-h cultures, 18-h treatment with IL-5 or NGF did not affect C5a-induced leukotriene C4 generation or alter C5a induced intracellular Ca2+ concentration elevations. IL-3 and IL-5, but not NGF, induced Fc(epsilon)RIbeta mRNA expression and all three improved basophil viability in culture with a ranking of IL-3 > IL-5 > or = NGF. All three cytokines acutely activated the extracellular signal-regulated kinase pathway and the signaling elements that preceded extracellular signal-regulated kinase and cytosolic phospholipase A2 phosphorylation, consistent with their redundant ability to acutely prime basophils. However, only IL-3 and IL-5 induced Janus kinase 2 and STAT5 phosphorylation. This pattern of signal element activation among the three cytokines most closely matched their ability to induce expression of Fc(epsilon)RIbeta mRNA. Induction of the sustained calcium signaling that follows overnight priming with IL-3 appeared to be related to the strength of the early signals activated by these cytokines but the relevant pathway required was not identified. None of the signaling patterns matched the ability of the cytokines to promote basophil survival. PMID- 11490017 TI - Eosinophil tissue recruitment to sites of allergic inflammation in the lung is platelet endothelial cell adhesion molecule independent. AB - Platelet endothelial cell adhesion molecule (PECAM or CD31) is a cell adhesion molecule expressed on circulating leukocytes and endothelial cells that plays an important role in mediating neutrophil and monocyte transendothelial migration in vivo. In this study, we investigated whether eosinophils, like neutrophils and monocytes, utilize PECAM for tissue recruitment to sites of allergic inflammation in vivo. Eosinophils express similar levels of PECAM as neutrophils as assessed by FACS analysis. RT-PCR studies demonstrate that eosinophils like neutrophils express the six extracellular domains of PECAM. Eosinophils exhibit homophilic binding to recombinant PECAM as assessed in a single-cell micropipette adhesion assay able to measure the biophysical strength of adhesion of eosinophils to recombinant PECAM. The strength of eosinophil adhesion to recombinant PECAM is the same as that of neutrophil binding to recombinant PECAM and can be inhibited with an anti-PECAM Ab. Although eosinophils express functional PECAM, anti-PECAM Abs did not inhibit bronchoalveolar lavage eosinophilia, lung eosinophilia, and airway hyperreactivity to methacholine in a mouse model of OVA-induced asthma in vivo. Thus, in contrast to studies that have demonstrated that neutrophil and monocyte tissue recruitment is PECAM dependent, these studies demonstrate that eosinophil tissue recruitment in vivo in this model is PECAM independent. PMID- 11490018 TI - Sensitized mast cells migrate toward the antigen: a response regulated by p38 mitogen-activated protein kinase and Rho-associated coiled-coil-forming protein kinase. AB - Although mast cells accumulate within the mucosal epithelial layer of patients with allergic rhinitis and bronchial asthma, the responsible chemotactic factors are undefined. We investigated whether mast cells sensitized with Ag-specific IgE migrate toward the Ag. MC/9 mast cells sensitized with anti-DNP IgE migrated toward DNP-conjugated human serum albumin. This migration was directional, and the degree was stronger than that induced by stem cell factor. IL-3 and stem cell factor-dependent cultured mast cells derived from mouse bone marrow also migrated toward the Ag. Subsequent migration mediated by the Fc(epsilon)RI was significantly inhibited by incubating the cells with Y-27632, a Rho-associated coiled-coil-forming protein kinase inhibitor, or with SB203580, a p38 mitogen activated protein kinase (MAPK) inhibitor. Both p38 MAPK and MAPK-activated protein kinase (MAPKAPK)2 were activated following Fc(epsilon)RI aggregation, and activation of MAPKAPK2 was almost completely inhibited by 10 microM SB203580. Wortmannin or a low concentration of SB203580 partially inhibited MAPKAPK2, but did not block mast cell migration. In contrast, Y-27632 did not affect the activation of MAPKAPK2. These results indicate that Ag works not only as a stimulant for allergic mediators from IgE-sensitized mast cells, but also as a chemotactic factor for mast cells. Both p38 MAPK activation and Rho-dependent activation of Rho-associated coiled-coil-forming protein kinase may be required for Fc(epsilon)RI-mediated cell migration. PMID- 11490019 TI - C5b-9 terminal complement complex protects oligodendrocytes from death by regulating Bad through phosphatidylinositol 3-kinase/Akt pathway. AB - Apoptosis of oligodendrocytes is induced by serum growth factor deprivation. We showed that oligodendrocytes and progenitor cells respond to serum withdrawal by a rapid decline of Bcl-2 mRNA expression and caspase-3-dependent apoptotic death. Sublytic assembly of membrane-inserted terminal complement complexes consisting of C5b, C6, C7, C8, and C9 proteins (C5b-9) inhibits caspase-3 activation and apoptotic death of oligodendrocytes. In this study, we examined an involvement of the mitochondria in oligodendrocyte apoptosis and the role of C5b-9 on this process. Decreased phosphatidylinositol 3-kinase and Akt activities occurred in association with cytochrome c release and caspase-9 activation when cells were placed in defined medium. C5b-9 inhibited the mitochondrial pathway of apoptosis in oligodendrocytes, as shown by decreased cytochrome c release and inhibition of caspase-9 activation. Phosphatidylinositol 3-phosphate kinase and Akt activities were also induced by C5b-9, and the phosphatidylinositol 3-phosphate kinase inhibitor LY294002 reversed the protective effect of C5b-9. Phosphatidylinositol 3-phosphate kinase activity was also responsible for the phosphorylation of Bad at Ser112 and Ser136. This phosphorylation resulted in dissociation of Bad from the Bad/Bcl-xL complex in a G(i)alpha-dependent manner. The mitochondrial pathway of oligodendrocyte apoptosis is, therefore, inhibited by C5b-9 through post translational regulation of Bad. This mechanism may be involved in the promotion of oligodendrocyte survival in inflammatory demyelinating disorders affecting the CNS. PMID- 11490020 TI - Up-regulation of IL-10R1 expression is required to render human neutrophils fully responsive to IL-10. AB - We have recently shown that IL-10 fails to trigger Stat3 and Stat1 tyrosine phosphorylation in freshly isolated human neutrophils. In this study, we report that IL-10 can nonetheless induce Stat3 tyrosine phosphorylation and the binding of Stat1 and Stat3 to the IFN-gamma response region or the high-affinity synthetic derivative of the c-sis-inducible element in neutrophils that have been cultured for at least 3 h with LPS. Similarly, the ability of IL-10 to up regulate suppressor of cytokine signaling (SOCS)-3 mRNA was dramatically enhanced in cultured neutrophils and, as a result, translated into the SOCS-3 protein. Since neutrophils' acquisition of responsiveness to IL-10 required de novo protein synthesis, we assessed whether expression of IL-10R1 or IL-10R2 was modulated in cultured neutrophils. We detected constitutive IL-10R1 mRNA and protein expression in circulating neutrophils, at levels which were much lower than those observed in autologous monocytes or lymphocytes. In contrast, IL-10R2 expression was comparable in both cell types. However, IL-10R1 (but not IL-10R2) mRNA and protein expression was substantially increased in neutrophils stimulated by LPS. The ability of IL-10 to activate Stat3 tyrosine phosphorylation and SOCS 3 synthesis and to regulate IL-1 receptor antagonist and macrophage-inflammatory protein 1beta release in LPS-treated neutrophils correlated with this increased IL-10R1 expression, and was abolished by neutralizing anti-IL-10R1 and anti-IL 10R2 Abs. Our results demonstrate that the capacity of neutrophils to respond to IL-10, as assessed by Stat3 tyrosine phosphorylation, SOCS-3 expression, and modulation of cytokine production, is very dependent on the level of expression of IL-10R1. PMID- 11490021 TI - Real-time imaging of vascular endothelial-cadherin during leukocyte transmigration across endothelium. AB - Vascular endothelial-cadherin (VE-cadherin) is a component of the adherens junctions of endothelial cells whose role in endothelial transmigration of leukocytes has been controversial. Using a VE-cadherin/green fluorescent protein fusion construct (VEcadGFP) that mimics the native molecule, we visualized alterations in endothelial junctional structure in real time during transmigration of human neutrophils and monocytes in an in vitro flow model. We observed abundant transmigration occurring exclusively at the cell borders (paracellularly). Surprisingly, transmigration occurred both through de novo formation of transient gaps in VEcadGFP junctional distribution, and also through preexisting gaps. De novo gaps 4-6 microm in size were formed after a leukocyte arrived at a junction, whereas preexisting gaps were present even before the leukocyte had interacted with the endothelial cells contributing to a junction. Gaps rapidly resealed within 5 min after leukocyte transmigration. Migrating leukocytes appeared to push aside VEcadGFP in the plane of the junction, and this displaced material subsequently diffused back to refill the junction. To our knowledge, this is the first example where molecular events at the lateral junction have been tracked in real time during transmigration. PMID- 11490022 TI - Regulation of class II expression in monocytic cells after HIV-1 infection. AB - Human macrophage hybridoma cells were used to study HLA-DR expression after HIV-1 infection. HLA-DR surface expression was lost 2 wk after infection that was associated with decreased mRNA transcription. Transfecting HLA-DR-alpha and HLA DR-beta cDNA driven by a nonphysiological CMV promoter restored expression, suggesting that regulatory DNA-binding proteins may be affected by HIV-1 infection. There was no protein binding to conserved class II DNA elements (W/Z/S box, X-1 and X-2 boxes, and Y box) in a HIV-1-infected human macrophage hybridoma cell line, 43(HIV), and in primary monocytes that lost HLA-DR expression after HIV-1(BaL) infection. PCR analysis of the HIV-1-infected cells that lost HLA-DR expression revealed mRNA for W/Z/S (RFX-5), X-1 (RFX-5), X-2 (hX-2BP), and one Y box DNA-binding protein (NF-YB), and CIITA, a non-DNA-binding protein necessary for class II transcription. There was no mRNA for the Y box-binding protein, NF YA. However, HLA-DR expression could be restored by transfection with NF-YA driven by a CMV promoter, although HLA-DR failed to localize in either the late endosomes, lysosomes, or acidic compartments. This was associated with a loss of class II-associated invariant chain peptide and leupeptin-induced protein in the 43(HIV) cells. To address this further, non-HIV-1-infected 43 cells were infected with vaccinia virus containing HIV-1 gag, nef, pol, and env proteins. HLA-DR failed to localize in neither the late endosomes, lysosomes, or acidic compartments in the vaccinia-infected cells containing HIV-1 env protein. HIV-1 appears to have multiple effects on class II expression in monocytic cells that may contribute to the immune defects seen in HIV-1-infected patients. PMID- 11490023 TI - Catalytic subunit of protein kinase A is an interacting partner of the inflammation-responsive transcription factor serum amyloid A-activating factor-1. AB - Serum amyloid A-activating factor-1 (SAF-1) is a zinc finger transcription factor that is activated by many mediators of inflammation including IL-1, IL-6, and bacterial LPS. However, the mechanism of activation is not fully understood. To identify possible activation partners for SAF-1, we used a yeast two-hybrid system that detected interaction between the catalytic subunit of cyclic AMP dependent protein kinase (PKA-Calpha) and SAF-1. Immunofluorescence and combined immunoprecipitation-Western blot analyses revealed colocalization and interaction between SAF-1 and PKA-Calpha. In vivo evidence of SAF-1 and PKA-Calpha interaction was further revealed by coimmunoprecipitation of these two proteins in cAMP-activated liver cells. We further show that SAF-1 is phosphorylated in vitro by PKA-Calpha and that addition of cAMP markedly induces in vivo phosphorylation of SAF-1 and transcription of SAF-regulated reporter genes. These results showed that SAF1-PKA-Calpha interaction is involved in functional activation of SAF-1. PMID- 11490024 TI - A critical role for mouse CXC chemokine(s) in pulmonary neutrophilia during Th type 1-dependent airway inflammation. AB - Ag-specific Th1 and Th2 cells have been demonstrated to play a critical role in the induction of allergic diseases. Here we have investigated the precise mechanisms of Th1-induced airway inflammation. Airway inflammation was induced in BALB/c mice by transfer of freshly induced OVA-specific Th1 or Th2 cells followed by OVA inhalation. In this model, both Th1 and Th2 cells induced airway inflammation. The former induced neutrophilia in airways, whereas the latter induced eosinophilia. Moreover, we found that Th1 cells induced more severe airway hyperresponsiveness (AHR) than Th2 cells. The eosinophilia induced by Th2 cell infusion was almost completely blocked by administration of anti-IL-5 mAb, but not anti-IL-4 mAb. In contrast, Th1-induced AHR and pulmonary neutrophilia were inhibited by the administration of anti-human IL-8R Ab, which blocks the function of mouse CXC chemokine(s). These findings reveal a critical role of mouse CXC chemokine(s) in Th1-dependent pulmonary neutrophilia and AHR. PMID- 11490025 TI - The role of alpha beta+ T cells and homeostatic T cell proliferation in Y chromosome-associated murine lupus. AB - Male BXSB mice develop an early life, severe lupus-like disease largely attributed to an undefined Y-chromosome-associated autoimmunity accelerator, termed YAA: Although the exact disease pathogenesis is uncertain, indirect evidence suggests that T cells play an important role in the male BXSB disease. We have developed TCR alpha-chain gene-deleted BXSB mice to directly examine the role of alphabeta+ T cells and the mode by which Yaa promotes disease in this strain. All disease parameters, including hypergammaglobulinemia, autoantibody production, glomerulonephritis, and the unique monocytosis of BXSB males, were severely reduced or absent in the alphabeta+ T cell-deficient mice. Adoptively transferred CD4+ T cells of either male or female BXSB origin showed equal homeostatic proliferation in alphabeta+ T cell-deficient male recipients. Moreover, deficient male mice eventually developed equally severe lupus-like disease after adoptive transfer and homeostatic expansion of T cells from wild type BXSB males or females. The results directly demonstrate that the Yaa mediated disease requires alphabeta+ T cells that are not, in themselves, abnormal in either composition or properties, but are engaged by a Yaa-encoded abnormality in a non-T cell component. In addition, homeostatic anti-self proliferation of mature T cells derived from a small number of precursors can induce systemic autoimmunity in an appropriate background. PMID- 11490026 TI - Increased frequency of pre-germinal center B cells and plasma cell precursors in the blood of children with systemic lupus erythematosus. AB - We have analyzed the blood B cell subpopulations of children with systemic lupus erythematosus (SLE) and healthy controls. We found that the normal recirculating mature B cell pool is composed of four subsets: conventional naive and memory B cells, a novel B cell subset with pregerminal center phenotype (IgD(+)CD38(+)centerin(+)), and a plasma cell precursor subset (CD20( )CD19(+/low)CD27(+/++) CD38(++)). In SLE patients, naive and memory B cells (CD20(+)CD38(-)) are approximately 90% reduced, whereas oligoclonal plasma cell precursors are 3-fold expanded, independently of disease activity and modality of therapy. Pregerminal center cells in SLE are decreased to a lesser extent than conventional B cells, and therefore represent the predominant blood B cell subset in a number of patients. Thus, SLE is associated with major blood B cell subset alterations. PMID- 11490027 TI - Fas/Fas ligand deficiency results in altered localization of anti-double-stranded DNA B cells and dendritic cells. AB - Autoantibodies directed against dsDNA are found in patients with systemic lupus erythematosus as well as in mice functionally deficient in either Fas or Fas ligand (FasL) (lpr/lpr or gld/gld mice). Previously, an IgH chain transgene has been used to track anti-dsDNA B cells in both nonautoimmune BALB/c mice, in which autoreactive B cells are held in check, and MRL-lpr/lpr mice, in which autoantibodies are produced. In this study, we have isolated the Fas/FasL mutations away from the autoimmune-prone MRL background, and we show that anti dsDNA B cells in Fas/FasL-deficient BALB/c mice are no longer follicularly excluded, and they produce autoantibodies. Strikingly, this is accompanied by alterations in the frequency and localization of dendritic cells as well as a global increase in CD4 T cell activation. Notably, as opposed to MRL-lpr/lpr mice, BALB-lpr/lpr mice show no appreciable kidney pathology. Thus, while some aspects of autoimmune pathology (e.g., nephritis) rely on the interaction of the MRL background with the lpr mutation, mutations in Fas/FasL alone are sufficient to alter the fate of anti-dsDNA B cells, dendritic cells, and T cells. PMID- 11490028 TI - Adoptive immunotherapy of experimental autoimmune encephalomyelitis via T cell delivery of the IL-12 p40 subunit. AB - CD4+ T cells are believed to play a central role in the initiation and perpetuation of autoimmune diseases such as multiple sclerosis. In the murine model for multiple sclerosis, experimental autoimmune encephalomyelitis, pathogenic T cells exhibit a Th1-like phenotype characterized by heightened expression of proinflammatory cytokines. Systemic administration of "regulatory" cytokines, which serve to counter Th1 effects, has been shown to ameliorate autoimmune responses. However, the inherent problems of nonspecific toxicity limit the usefulness of systemic cytokine delivery as a potential therapy. Therefore, we used the site-specific trafficking properties of autoantigen reactive CD4+ T cells to develop an adoptive immunotherapy protocol that provided local delivery of a Th1 cytokine antagonist, the p40 subunit of IL-12. In vitro analysis demonstrated that IL-12 p40 suppressed IFN-gamma production in developing and effector Th1 populations, indicating its potential to modulate Th1 promoted inflammation. We have previously demonstrated that transduction of myelin basic protein-specific CD4+ T cells with pGC retroviral vectors can result in efficient and stable transgene expression. Therefore, we adoptively transferred myelin basic protein-specific CD4+ T cells transduced to express IL 12 p40 into mice immunized to develop experimental autoimmune encephalomyelitis and demonstrated a significant reduction in clinical disease. In vivo tracking of bioluminescent lymphocytes, transduced to express luciferase, using low-light imaging cameras demonstrated that transduced CD4+ T cells trafficked to the central nervous system, where histological analysis confirmed long-term transgene expression. These studies have demonstrated that retrovirally transduced autoantigen-specific CD4+ T cells inhibited inflammation and promoted immunotherapy of autoimmune disorders. PMID- 11490029 TI - Glomerulonephritis induced by recombinant collagen IV alpha 3 chain noncollagen domain 1 is not associated with glomerular basement membrane antibody: a potential T cell-mediated mechanism. AB - Glomerulonephritis is believed to result commonly from Ab-mediated glomerular injury. However, Ab-associated mechanisms alone cannot explain many cases of human glomerulonephritis. We developed a rat model of human anti-glomerular basement membrane (GBM) disease to investigate T cell and Ab response, and their associations with the disease. A single immunization of highly denatured recombinant mouse collagen IV alpha3 chain noncollagen domain 1 (rCol4alpha3NC1) induced severe glomerulonephritis in 100% of Wistar Kyoto rats, 33% of which died of this disease around day 35 postimmunization. The renal pathology demonstrated widespread glomerular damage and a mononuclear cell infiltration within the interstitial tissue. T cells from immunized rats responded not only to rCol4alpha3NC1, but also to isolated rat GBM. Sera Abs to rCol4alpha3NC1 were detectable in 100% of the rats, but only 20% of the rats had low levels of Ab to isolated rat GBM by Western blot, and none by immunofluorescence. Furthermore, IgG/M binding to or C3 deposition on endogenous GBM in immunized rats were not detected in most of the experimental rats, and showed no statistical correlation with disease severity. Additionally, no electronic dense deposition in the glomeruli was detected in all rats. Those data revealed a disassociation between the disease and anti-GBM Ab. T cell-mediated mechanisms, which are currently under our investigation, may be responsible for the glomerular disease. PMID- 11490030 TI - Resistance to xenobiotic-induced autoimmunity maps to chromosome 1. AB - Although evidence indicates that environmental factors play a major role in precipitating systemic autoimmunity in genetically susceptible individuals, little is known about the mechanisms involved. Certain heavy metals, such as mercury, are potent environmental immunostimulants that produce a number of immunopathologic sequelae, including lymphoproliferation, hypergammaglobulinemia, and overt systemic autoimmunity. Predisposition to such metal-induced immunopathology has been shown to be influenced by both MHC and non-MHC genes, as well as susceptibility to spontaneous lupus, in mice and other experimental animals. Among the various mouse strains examined to date, the DBA/2 appears to uniquely lack susceptibility to mercury-induced autoimmunity (HgIA), despite expressing a susceptible H-2 haplotype (H-2d). To define the genetic basis for this trait, two genome-wide scans were conducted using F2 intercrosses of the DBA/2 strain with either the SJL or NZB strains, both of which are highly susceptible to HgIA. A single major quantitative trait locus on chromosome 1, designated Hmr1, was shown to be common to both crosses and encompassed a region containing several lupus susceptibility loci. Hmr1 was linked to glomerular immune complex deposits and not autoantibody production, suggesting that DBA/2 resistance to HgIA may primarily involve the later stages of disease pathogenesis. Identification and characterization of susceptibility/resistance genes and mechanisms relevant to the immunopathogenesis of mercury-induced autoimmunity should provide important insights into the pathogenesis of autoimmunity and may reveal novel targets for intervention. PMID- 11490031 TI - Inhibition of autoimmune diabetes in nonobese diabetic mice by transgenic restoration of H2-E MHC class II expression: additive, but unequal, involvement of multiple APC subtypes. AB - Transgenic restoration of normally absent H2-E MHC class II molecules on APC dominantly inhibits T cell-mediated autoimmune diabetes (IDDM) in nonobese diabetic (NOD) mice. We analyzed the minimal requirements for transgenic H2-E expression on APC subtypes (B lymphocytes vs macrophages/dendritic cells (DC)) to inhibit IDDM. This issue was addressed through the use of NOD stocks transgenically expressing high levels of H2-E and/or made genetically deficient in B lymphocytes in a series of genetic intercross and bone marrow/lymphocyte chimera experiments. Standard (H2-E(null)) NOD B lymphocytes exert a pathogenic function(s) necessary for IDDM. However, IDDM was inhibited in mixed chimeras where H2-E was solely expressed on all B lymphocytes. Interestingly, this resistance was abrogated when even a minority of standard NOD H2-E(null) B lymphocytes were also present. In contrast, in NOD chimeras where H2-E expression was solely limited to approximately half the macrophages/DC, an active immunoregulatory process was induced that inhibited IDDM. Introduction of a disrupted IL-4 gene into the NOD-H2-E transgenic stock demonstrated that induction of this Th2 cytokine does not represent the IDDM protective immunoregulatory process mediated by H2-E expression. In conclusion, high numbers of multiple subtypes of APC must express H2-E MHC class II molecules to additively inhibit IDDM in NOD mice. This raises a high threshold for success in future intervention protocols designed to inhibit IDDM by introduction of putatively protective MHC molecules into hemopoietic precursors of APC. PMID- 11490032 TI - Down-regulation of IgE and IgG4 antibodies to tetanus toxoid and diphtheria toxoid by covaccination with cellular Bordetella pertussis vaccine. AB - Pertussis (P) toxin acts as adjuvant for IgE formation against simultaneously administered Ags in animal models. P vaccination may also have an adjuvant impact on IgE formation against coadministered diphtheria (D) and tetanus (T) Ags in humans. Sera of 103 D-T-P-immunized and 319 D-T-immunized children aged 2 years were analyzed for IgE, IgG4, and IgG to D and T (radioallergosorbent test), total IgE and IgE against common inhalant allergens (CAP radioallergosorbent test fluoroenzyme immunoassay). Fewer D-T-P- than D-T-immunized children had sera positive for T-IgE (12.6 vs 53.6%, p < 0.001), T-IgG4 (71.6 vs 89.2%, p < 0.001), D-IgE (31.0 vs 70.5%, p < 0.001), and D-IgG4 (85.2 vs 93.4%, p = 0.039). Suppression of T-IgE was not dependent on the cutoff chosen for a positive test result, but was dependent on the proportion of D-T immunizations given with P. The risk for sensitization to common environmental allergens did not differ (odds ratio 0.953, 95% confidence interval 0.815-1.114). No significant differences between D-T- and D-T-P-immunized children were found with regard to T-IgG or D IgG. In summary, IgE and IgG4 (but not IgG) serum levels to coadministered D- and T-Ags are suppressed among P-immunized children as compared with nonimmunized children. These results suggest that the presence of a microbial product during Ag exposure can down-regulate an IgE/IgG4 response in humans. PMID- 11490033 TI - Effects of home environment, socioeconomic status, and health status on cognitive functioning in children with HIV-1 infection. AB - OBJECTIVE: To investigate the effects of the home environment, socioeconomic status (SES), and health status on cognitive functioning in a sample of children with HIV-1 infection in a cross-sectional study. METHODS: Forty-three caregivers and their children (2.5 to 12 years) participated. Caregivers completed two self report measures of the home environment that included questions regarding the organization of the environment, play materials, parental involvement, variety of stimulation, and parental attitudes toward the provision of a cognitively stimulating environment. Cognitive functioning was assessed using a standardized intelligence (IQ) test. Children's medical charts were reviewed for HIV-1 classification status (CDC, 1994), CD4 cell counts, and current medication. RESULTS: This study revealed two primary findings. First, measures of the home environment mediated the association between SES and child IQ. Second, measures of the home environment had a stronger association with child IQ during the advanced stages of disease than earlier stages of disease. CONCLUSIONS: The home environment is associated with cognitive functioning among children with HIV-1 infection. Moreover, interventions aimed at enhancing the quality of the home environment may have a positive impact on these children's cognitive development. PMID- 11490034 TI - Psychological adjustment in caregivers of school-age children infected with HIV: stress, coping, and family factors. AB - OBJECTIVE: To assess for significant differences in psychological functioning between caregivers of HIV-infected children and caregivers of healthy children, and to examine the utility of applying a stress and coping model to caregivers of children with HIV disease. METHODS: Participants included caregivers of HIV infected children (n = 36) and caregivers of a demographically matched control group of healthy children (n = 32). During their child's pediatric clinic visits, caregivers completed measures of psychological adjustment, stress, coping style, and family resources and support. They also completed a measure of their child's psychological adjustment. RESULTS: Caregiver psychological distress scores did not significantly differ between the HIV and control groups, and clinically significant rates of psychological distress were reported by more than a third of caregivers in both groups. Hierarchical multiple regression analyses revealed that independent of their child's illness status, stress and coping style were significant predictors of caregiver's psychological adjustment. In addition, caregiver psychological distress was a significant predictor of children's maladjustment. CONCLUSIONS: High rates of psychological distress were observed in caregivers of children with HIV disease; however, similarly high rates of psychological adjustment problems were found in caregivers of healthy children. Caregivers who reported high levels of daily stress and emotion-focused coping styles tended to report more psychological distress. Further, caregivers who reported more psychological distress also reported more internalizing and more externalizing behavior problems in their children, regardless of the child's illness status. These findings reflect the impact of poverty and environmental stress on caregivers' adjustment. PMID- 11490035 TI - Predictors of psychological adjustment in school-age children infected with HIV. AB - OBJECTIVE: To assess for significant differences in psychological functioning between HIV-infected children and a demographically matched healthy control group and to examine the utility of applying a stress and coping model to children with HIV disease. METHODS: Participants included HIV-infected children (ages 6-16) and their caregivers (n = 36) and a control group of healthy children and their caregivers (n = 32). During routine clinic visits, children completed measures of psychological adjustment, health locus of control, and coping style, and caregivers completed measures of their own and their child's psychological adjustment. RESULTS: Caregiver-reported and child self-reported psychological adjustment scores did not significantly differ between the HIV and control groups, with the exception of significantly more internalizing behavior problems reported in the control group. Hierarchical multiple regression analyses revealed that the stress and coping model accounted for 36% of the variance in HIV infected children's self-reported psychological adjustment. In addition, child age and coping style were significant predictors of child self-reported psychological adjustment, but not of caregiver-reported child adjustment. CONCLUSIONS: Approximately 25% of children with HIV disease exhibited clinically significant emotional or behavioral problems; however, even higher rates of psychological adjustment problems were found in healthy children. Children with HIV disease who have not been told their diagnosis and children who endorse more emotion-focused coping strategies tend to exhibit more psychological adjustment problems. PMID- 11490036 TI - Brief report: fathering a child living with HIV/AIDS: psychosocial adjustment and parenting stress. AB - OBJECTIVE: To examine the psychosocial stressors experienced by fathers of children diagnosed with HIV/AIDS. METHODS: Thirty-one fathers whose children (ages 6 to 19) were participating in pediatric HIV clinical trials completed self report measures of parenting stress, psychological distress, and need for psychosocial services. RESULTS: Over half of this sample experienced significantly elevated levels of both parenting stress and psychological distress compared to standardized norms. Ninety-seven percent of these men reported the need for services including gender-specific support groups, assistance with discipline, disease management, and assistance with planning for the future. CONCLUSIONS: Elevated levels of parenting stress and psychological distress in fathers of children living with HIV suggest the need for additional psychological intervention in this population. PMID- 11490037 TI - Commentary: coping over the long haul: understanding and supporting children and families affected by HIV disease. PMID- 11490038 TI - Commentary: HIV infection and family processes: toward the next generation of research. PMID- 11490039 TI - Children's expectations and memories of acute distress: short- and long-term efficacy of pain management interventions. AB - OBJECTIVE: To examine the effect of psychologic and pharmacologic interventions on children's expectations and 6-month recollections of painful procedures. METHODS: A repeated measures design allowed examination of 22 fourth graders' expectations, experiences, and memories of distress across three conditions (typical care, distraction, topical anesthetic) for a three-injection vaccination series. All participants were African American and from urban, low-income families. RESULTS: Across conditions, children's expectations of distress were significantly higher than their experience of distress. Distress ratings did not differ among conditions prior to or immediately following the injections; however, children later recalled that the treatment conditions were superior to control for distress relief. Analyses of recall accuracy suggest that the interventions buffered the children from forming negative recollections that occurred with typical care. CONCLUSIONS: Children have negative expectations prior to a procedure despite knowing that a distress management intervention will be employed. However, interventions may thwart the development of negative memories of distress. PMID- 11490040 TI - Mothers' representations of relationships with their children: relations with mother characteristics and feeding sensitivity. AB - OBJECTIVE: To examine parenting representations and feeding interactions of mothers and their children with cerebral palsy (CP) and the extent to which mothers' representations predict their feeding behavior beyond other mother and child characteristics. METHODS: Fifty-eight mothers of children with mild to severe CP ages 16 to 52 months were interviewed with an adapted form of the Parent Development Interview (PDI). Correlation and regression analyses examined relations between representations (compliance with parental requests, achievement, secure base, enmeshment, worry about the child's future, and emotional pain), demographic characteristics, diagnostic severity, and developmental status. RESULTS: Mothers with more compliance-related concerns showed less sensitivity, acceptance, and delight during feeding. Mothers experiencing more emotional pain displayed more hostility. Mothers reporting worries about the child displayed sensitivity and delight. Representations of compliance-related experiences and worry about the child's future accounted for significant increments in explained variance in mothers' feeding behavior, after we controlled for children's skills and abilities. CONCLUSIONS: Findings suggest maternal representations of relationships are associated with caregiving behavior for mothers of children with CP apart from other child and maternal characteristics and may be a useful focus for research and practice related to parenting children with special needs. PMID- 11490041 TI - Assisted suicide: a few answers--many questions. PMID- 11490042 TI - A comparison of characteristics of Kevorkian euthanasia cases and physician assisted suicides in Oregon. AB - PURPOSE: The sociodemographic and clinical characteristics of Kevorkian euthanasia cases were compared with Oregon physician-assisted suicide (PAS) cases and U.S. mortality data. DESIGN AND METHODS: Two hundred variables were coded from medical examiner reports on all 69 Kevorkian euthanasia cases who died and were autopsied by the Oakland County Medical Examiner. Data on the 43 Oregon PAS cases in the first two years and U.S. mortality data were obtained from published sources. RESULTS: Only 25% of patients euthanized by Kevorkian were terminally ill as compared to 100% of Oregon PAS cases. PAS cases were significantly more likely to have cancer (72%) than euthanasia cases (29%). Women and those who were divorced or had never married were significantly more likely to seek euthanasia than would have been predicted by national mortality statistics. IMPLICATIONS: Gender and marital status appeared to influence decisions to seek an assisted death, and research on the role of these factors in end-of-life decision making is merited. PMID- 11490044 TI - The stress process of family caregiving in institutional settings. AB - PURPOSE: This study adapts the Stress Process Model (SPM) of family caregiving to examine the predictors of depression in a sample of caregivers (n = 133) with demented relatives residing in suburban skilled nursing facilities. DESIGN AND METHODS: We interviewed family caregivers of family members residing in skilled nursing homes using a variety of measures to assess primary stressors, secondary strain, nursing home stressors, and caregiver depression. We used block-wise regression analyses to determine the predictors of caregiver depression. RESULTS: Results indicated that positive resident adjustment to placement was best predicted by the closeness of the resident-caregiver relationship and nursing home stressors. Caregiver strain, resident adjustment, and nursing home stressors best predicted caregiver adjustment. In turn, the best predictors of caregiver depression included caregiver age, caregiver adjustment to the nursing home, and nursing home stressors. IMPLICATIONS: These results suggest that caregiver depression is closely linked to how well both the resident and caregiver adjust to the nursing home environment. Results also indicate that by broadening the SPM to include stressors common to the nursing home experience, researchers will be able to understand more clearly the specific components of the stress process that may lead to depression in family caregivers of persons living in nursing homes. PMID- 11490043 TI - Come talk with me: improving communication between nursing assistants and nursing home residents during care routines. AB - PURPOSE: We examined the effects of communication skills training and the use of memory books by certified nursing assistants (CNAs) on verbal interactions between CNAs (n = 64) and nursing home residents (n = 67) during care routines. DESIGN AND METHODS: CNAs were taught to use communication skills and memory books during their interactions with residents with moderate cognitive impairments and intact communication abilities. A staff motivational system was used to encourage performance and maintenance of these skills. Formal measures of treatment implementation were included. RESULTS: Results were compared with those for participants on no-treatment control units. Trained CNAs talked more, used positive statements more frequently, and tended to increase the number of specific instructions given to residents. Changes in staff behavior did not result in an increase in total time giving care to residents. Maintenance of CNA behavior change was found 2 months after research staff exited the facility. Although an increase was found in positive verbal interactions between CNAs and residents on intervention units, other changes in resident communication were absent. IMPLICATIONS: Nursing staff can be trained to improve and maintain communication skills during care without increasing the amount of time delivering care. The methodological advantages of including measures to assess treatment implementation are discussed. PMID- 11490045 TI - Home- and community-based long-term care: lessons from Denmark. AB - PURPOSE: Denmark is cited as a model in the development of home- and community based systems for the frail elderly population. We examined the results of this natural experiment and considered implications for U.S. policy. DESIGN AND METHODS: We used international comparative policy analysis, including site visits and semistructured interviews with Danish leadership in conjunction with a review of published literature, reports, and administrative data from Denmark and the United States. RESULTS: After 12 years of implementing integrated systems for home- and community-based services in 275 municipalities, growth in Danish long term care expenditures has leveled off; expenditures appear to be decreasing for the over-80 population and have dropped as a percentage of the gross domestic product. Access to and quality of long-term care services appear to remain generally satisfactory. During this period, comparable expenditures in the United States have increased, and deficits in access and quality persist. IMPLICATIONS: These findings should be of interest to state and federal policy makers considering strategies to reduce the rate of growth in Medicaid and Medicare expenditures for elders and to expand home- and community-based services. PMID- 11490046 TI - Judging outcomes in psychosocial interventions for dementia caregivers: the problem of treatment implementation. AB - PURPOSE: In published dementia caregiver intervention research, there is widespread failure to measure the level at which treatment was implemented as intended, thereby introducing threats to internal and external validity. The purpose of this article is to discuss the importance of inducing and assessing treatment implementation (TI) strategies in caregiving trials and to propose Lichstein's TI model as a potential guide. DESIGN AND METHODS: The efforts of a large cooperative research study of caregiving interventions, Resources for Enhancing Alzheimer's Caregiver Health (REACH), illustrates induction and assessment of the three components of TI: delivery, receipt, and enactment. RESULTS: The approaches taken in REACH vary with the intervention protocols and include using treatment manuals, training and certification of interventionists, and continuous monitoring of actual implementation. IMPLICATIONS: Investigation and description of treatment process variables allows researchers to understand which aspects of the intervention are responsible for therapeutic change, potentially resulting in development of more efficacious and efficient interventions. PMID- 11490047 TI - Gender differences in cardiovascular response to dementia caregiving. AB - PURPOSE: This study examined gender differences in cardiovascular responses to laboratory-based stress, as well as in ambulatory hemodynamic (i.e., blood pressure and heart rate) functioning among caregivers of persons with dementia. DESIGN & METHODS: Participants were 25 men and 25 women caregivers, matched on age, type of care recipient's dementia, and relationship to the care recipient. After cardiovascular reactivity to a laboratory-based caregiving stressor was assessed, the ambulatory hemodynamic functioning levels of caregivers were measured in caregivers' natural environments. RESULTS: Female caregivers displayed greater systolic and diastolic blood pressure reactivity to a laboratory-based stress task (i.e., discussing caregiving difficulties) compared with male caregivers (p < or =.01). In contrast, no gender differences were found for ambulatory hemodynamic functioning when aggregated overall or when in the presence of the care recipient. IMPLICATIONS: Laboratory-based findings suggest that female caregivers experience greater blood pressure reactivity to caregiving related stress than do male caregivers. However, these laboratory-based gender differences may not generalize to differences in hemodynamic functioning in caregivers' daily lives. PMID- 11490048 TI - Explaining the role of sex on disability: a population-based study. AB - PURPOSE: There is no clear consensus about how sex influences prevalent disability and through what mechanisms. We investigated whether sex had a direct independent effect on disability or whether sex had an interactive effect on the relationship between chronic diseases/conditions and disability, and whether these effects differed in middle-aged versus older adults. DESIGN AND METHODS: We used baseline data from two nationally representative health interview surveys, the Health and Retirement Study (HRS) and the Study of Asset and Health Dynamics Among the Oldest Old (AHEAD), and disability and covariate measures that were nearly identical in both surveys. Logistic regression models tested the contributions of diseases, impairments, and demographic and social characteristics on difficulties with prevalent activities of daily living (ADLs), mobility, and strength. RESULTS: Models demonstrated no direct sex effect for ADL disability in either age group after adjusting for key covariates. However, sex did exert an indirect effect on ADL disability in older adults via musculoskeletal conditions and depressive symptoms. In contrast, female sex remained strongly associated with mobility and strength disability in both age groups, net of covariates. Major interactions were also significant, including a female sex/body mass index (BMI) interaction for mobility difficulty and several sex-disease interactions for strength disability in the middle-aged group. IMPLICATIONS: The effect of sex on ADL difficulty is largely explained by social and health-related covariates in middle-aged and older adults. In contrast, the independent association of female sex with decreased strength and mobility in both groups cannot be explained by our models' social or health-related variables. In addition, the positive association of BMI with mobility difficulty is significantly worse for women than for men. PMID- 11490049 TI - Improving attitudes regarding the elderly population: the effects of information and reinforcement for change. AB - PURPOSE: Altering negative attitudes associated with ageism may be possible by giving people accurate information about older people in conjunction with reinforcement for change. DESIGN AND METHODS: Ninety-nine college students (35 men, 63 women; mean age = 20 years, SD = 2.78) participated in one of three groups: information only, information plus an innocuous discussion group, and information plus a reinforcement-to-change discussion group. The participants' attitudes toward elderly people were measured before, immediately after the intervention, and at a one-month follow-up. Changes in attitudes across groups and time were analyzed using analysis of variance (ANOVA) and t tests. RESULTS: Information alone produced initial improvements in attitudes in all groups; however, only the group members who received additional reinforcement for change maintained positive attitude changes at one-month follow-up. IMPLICATIONS: This study supports the premise that negative attitudes toward older people are amendable; however, the new attitude may be lost without reinforcement for change. PMID- 11490050 TI - Social network type and morale in old age. AB - PURPOSE: The aim of this research was to derive network types among an elderly population and to examine the relationship of network type to morale. DESIGN AND METHODS: Secondary analysis of data compiled by the Israeli Central Bureau of Statistics (n = 2,079) was employed, and network types were derived through K means cluster analysis. Respondents' morale scores were regressed on network types, controlling for background and health variables. RESULTS: Five network types were derived. Respondents in diverse or friends networks reported the highest morale; those in exclusively family or restricted networks had the lowest. Multivariate regression analysis underscored that certain network types were second among the study variables in predicting respondents' morale, preceded only by disability level (Adjusted R(2) =.41). IMPLICATIONS: Classification of network types allows consideration of the interpersonal environments of older people in relation to outcomes of interest. The relative effects on morale of elective versus obligated social ties, evident in the current analysis, is a case in point. PMID- 11490051 TI - Randomized clinical trial of a quality improvement intervention in nursing homes. AB - PURPOSE: The purpose of the study was to determine if simply providing nursing facilities with comparative quality performance information and education about quality improvement would improve clinical practices and subsequently improve resident outcomes, or if a stronger intervention, expert clinical consultation with nursing facility staff, is needed. DESIGN AND METHODS: Nursing facilities (n = 113) were randomly assigned to one of three groups: workshop and feedback reports only, workshop and feedback reports with clinical consultation, and control. Minimum Data Set (MDS) Quality Indicator (QI) feedback reports were prepared and sent quarterly to each facility in intervention groups for a year. Clinical consultation by a gerontological clinical nurse specialist (GCNS) was offered to those in the second group. RESULTS: With the exception of MDS QI 27 (little or no activity), no significant differences in resident assessment measures were detected between the groups of facilities. However, outcomes of residents in nursing homes that actually took advantage of the clinical consultation of the GCNS demonstrated trends in improvements in QIs measuring falls, behavioral symptoms, little or no activity, and pressure ulcers (overall and for low-risk residents). IMPLICATIONS: Simply providing comparative performance feedback is not enough to improve resident outcomes. It appears that only those nursing homes that sought the additional intensive support of the GCNS were able to effect enough change in clinical practice to improve resident outcomes significantly. PMID- 11490052 TI - Care-as-service, care-as-relating, care-as-comfort: understanding nursing home residents' definitions of quality. AB - PURPOSE: This study explored how nursing home residents define quality of care. DESIGN AND METHODS: Data were collected through in-depth interviews and were analyzed using grounded dimensional analysis. RESULTS: Residents defined quality in three ways: (a) Care-as-service residents focused on instrumental aspects of care. They assessed quality using the parameters of efficiency, competence, and value. (b) Care-as-relating residents emphasized the affective aspects of care, defining quality as care that demonstrated friendship and allowed them to show reciprocity with their caregivers. (c) Care-as-comfort residents defined quality as care that allowed them to maintain their physical comfort, a state that required minute and often repetitive adjustments in response to their bodily cues. IMPLICATIONS: Residents' perceptions of care quality have implications for long-term care practice. The integration of these perceptions into quality assurance instruments could improve the usefulness of tools designed to obtain resident input. PMID- 11490053 TI - The feasibility of using a telephone-administered survey for determining nutritional service needs of noninstitutionalized older adults in rural areas: time and costs. AB - PURPOSE: This study examined response, participation, time, and costs for a telephone-administered survey to obtain comprehensive information on general health, eating habits, living environment, and functional status from a sample of older persons in a rural North Carolina county. DESIGN AND METHODS: A probability sample of persons aged 60 years and older from the most recent electoral rolls were mailed a personalized letter, which was followed by telephone contact to recruit them into a contemporaneous survey that used a modified version of the Nutrition Screening Initiative's Level I and II screens. Time requirements and costs associated with the completion of surveys were calculated. RESULTS: Seventy six percent of the persons contacted by telephone (residents of 96% of county precincts) completed the survey. Because minority elders were more likely to lack a working telephone, they were underrepresented in the sample. With 555 calling attempts (58% of surveys completed on first attempt), we estimated a cost of $10.65 per completed survey. IMPLICATIONS: Telephone-administered surveying of older adults may be considered as an appropriate component of an overall community-based service strategy. The estimation of the constituents of nutritional risk, by geographic area, economic status, or ethnicity, may aid in providing estimates of service needs and procuring and allocating resources. Additional methods of data collection are necessary in order to target older persons without telephone service. PMID- 11490054 TI - Response to "Pain in U.S. nursing homes: validating a pain scale for the Minimum Data Set". PMID- 11490056 TI - [Health promotion. Concept development]. AB - The development of health promotion in theory and practice is presented-from the Ottawa Charter in 1986, to the community based health promotion programmes, as the vision for the 21 Century. The historical mile stones in the process of change and the conceptualisation of health promotion are discussed with reference to the World WHO Conferences and documents from these conferences. These events and documents have been vital as guidelines for member countries, both for implementation of community based programmes, as well as for healthy public policy and for training, especially concerning evaluation. The paper also discusses the main trends in research; definitions of principal concepts are highlighted, concerning planning, implementation and evaluation of health promotion programmes. PMID- 11490057 TI - [Evaluation of health promotion programmes]. AB - The paper contains a review of definitions of evaluation and discusses the need to evaluate health promotion programmes. The classification of types of evaluation is presented. It is aimed to create a common language of communication between evaluation and the common understanding of terms. The relation between evaluation of health promotion programmes and quality assurance, best practice and evidence based health promotion are discussed. PMID- 11490059 TI - [Health promotion indicators - selected model approaches]. AB - The aim of the paper is to present the problem, how to find the optimal set of indicators necessary in the different phases of planning, programming and evaluation of health promotion programmes. In health promotion the application of classic health measurements is limited, because they are not related to "positive health". Moreover, time perspective when effects of promotion could appear is difficult to define. Some model approaches are presented. The PRECEDE-PROCEED model is recognised as a robust framework in health promotion. This model provides a continuous series of steps and phases in planning, implementation and evaluation process. Some authors recommend already in the early phase of programming to draw a diagram showing a relationship between the objectives, activities, indicators and resources. In the process of evaluation it seems very important for health promotion initiatives to focus on the changes in human knowledge, attitudes and behaviours. For that reason models like KAP analysis (knowledge, attitudes, practice), HBM (Health Belief Model) and TTM (Transtheoretical Prochaska's model) are commonly applied. However, the- re is still a need to develop and choose indicators, which would consider environmental aspects of health promotion and link local actions with national health promotion policy. PMID- 11490058 TI - [Development of the concept of evaluation in educational programmes]. AB - The term evaluation has only recently found its place in educational terminology, in Poland. As in other domains, it has now become established, but even today there are studies in which such terms as control or assessment of teaching, without mentioning evaluation, are used. In the leading Polish education dictionary by Okon from 1984, there is no entry under evaluation. However, the concept of educational evaluation has been taken up in many pedagogic works by the 1990. The precise functions, the object of evaluation and the information needed should be well defined with respect to the evaluated programme. A ISO the criteria must be spelled out and the values of the planners and of the consumers ought to be concordant. This paper attempts to show the trends and the developing concepts in educational evaluation and their significant at the time of Poland's accession to the European Union. PMID- 11490060 TI - [Epidemiological studies in evaluation of effectiveness of health promotion programmes]. AB - Basic characteristics of modern epidemiology and basic characteristics of epidemiological study are presented. First of all, the description of the model of randomised controlled trial is taken into account, because of its importance for evaluation of effectiveness of treatment. Consequences of main elements of randomised controlled trial for scientific inference is explained and the possibility of application of these elements to evaluation of health promotion programmes is discussed. The main points in the discussion on the possibility of application of epidemiological methods to evaluation of health promotion programmes are mentioned. PMID- 11490061 TI - [Social context of health promotion program evaluation]. AB - The concept and methodology of health promotion programmes evaluation have much in common with the evaluation of all social programmes. Its goal is to improve the decision-making and to engage the community in this process. In this way, health promotion programmes gain procedures which analyse their effectiveness and acquire social validity. The role of evaluation studies is linked with the right to possess information about factors determining community participation in the managerial process. Designing and implementing evaluation research is in contradiction with the technocratic approach to problem solving and to planning and implementing programmes, which will improve community health. PMID- 11490062 TI - [Qualitative technics for social sciences--focus group interview for evaluation of health promotion programs and prevention of problems with adolescents]. AB - The paper presents basic information about focus group interviews (FGI). The presentation has been designed as a form of introduction for people with limited experience in applied social sciences. FGI are discussed in the context of their usefulness in evaluation of health promotion and prophylactics. The discussion concentrates on the following main topics; Specificity of qualitative studies (diversity of evaluation). FGI applicability. General interview methodology. Practical solutions for focus group research. PMID- 11490064 TI - [Evaluation as a component of breastfeeding promotion programme]. AB - Recent years have seen increased emphasis on appropriate development of public health intervention programmes with attention to evaluation. With reference to the breastfeeding promotion programme in Poland, the conceptual framework of evaluation development and implementation is presented. The use of analysis per formed for programme sustainability and progress and in the decision making process is underlined. PMID- 11490063 TI - [Principles of economic evaluation in intervention programmes]. AB - Since human and financial resources are limited there is a need to use them for justified health care projects. Cost-effectiveness is one, but not the only criterion which health care system may be asked to respect. This paper presents the principles of economic analysis as a tool of evaluation in intervention programmes. It describes the three most commonly used methods: cost effectiveness, cost-utility and cost-benefit analysis. An overview of concepts, calculation techniques and real applications of economic evaluation are presented. A review of the literature showed, that cost-utility and cost-benefit methods are developing very fast, and that is an increasing interest in knowledge and attitudes of decision-makers towards economic evaluation. PMID- 11490065 TI - [Evaluation of the health promoting school. Concepts and practice in Poland and Europe]. AB - The history of health promoting school movement in Poland, Europe and other regions in the last decade and the concept of this school was presented. The basic principles of process and outcome evaluation implemented in practice in the project "The health promoting school" (1992-1995) and the model (concept, methods, organisation) of evaluation of outcomes (at national and school level) of this project were described. The European Network of Health Promoting Schools focused on many initiatives during the past five years looking for effective and useful for schools methods of evaluation of health promoting school. Examples of indicators at international, national and school levels, focusing on dissemination, structures and impact, were presented. PMID- 11490066 TI - [Basic principles of evaluation of neural tube defects primary prevention programme]. AB - Neural Tube Defects (NTD) Primary Prevention Programme has been developed in Poland since 1997. The Programme directed by Professor Z. J. Brzezinski is based on an information and education campaign directed at health professionals, women in childbearing age and high school students. The Central Programme Office has been directing the development and implementation of the Programme through a network of provincial programmes. In the present paper the main principles of NTD primary prevention were presented as well as some principles of Programme building and evaluation. Due to the relatively short period of Programme implementation, it is too early to perform the full evaluation. However, some examples were presented including process evaluation and testing the main tools (questionnaires, school programme project). It was assumed that the effectiveness of NTD Primary Prevention Programme in Poland will be assessed by: 1) changes in women's knowledge, attitude and behaviour concerning folic acid supplementation, 2) changes in NTD mortality and morbidity rates, 3) changes in the amount of sold tablets containing 0.4 mg of folic acid. For each of these indicators some international results and Polish baseline results were discussed. PMID- 11490067 TI - [Evaluation in school anti-tobacco programmes]. AB - The National Research Institute of Mother and Child (NRIMC) school anti-tobacco programmes are presented. They are discussed with reference to the Centres for Disease Control and Prevention (CDC) guidelines and other effective school programmes to prevent tobacco use among children and young people. The general objectives of the programmes at different educational levels and the elaborated strategic recommendations are presented. The own conceptualised evaluation model of school anti-tobacco programmes is analysed, including process and outcome evaluation. The presented model clarifies concepts and procedures of planning the design and evaluation of the programme in its different stages. In our opinion, the NRIMC evaluation model can be applied in other heath promotion programmes. PMID- 11490068 TI - [Elaboration and evaluation of mental health promotion programmes exemplified by a primary prevention alcohol programme]. AB - The article's aim is a presentation of the process a/elaboration and evaluation of the designed mental health prevention programmes. Keeping the logic of this process (from overall goal setting to outcome assessment) increases the chance that resources for preventive and research activities will be used in the most favourable way. The process starting point is the establishment of the general goal of the programme. At this stage the clarification how the term "mental health promotion" and prevention activities are understood is crucial. The definition choice determines the area of activity and points out the target group. It is important to base the programme on solid theoretical and empirical evidence. This implies the necessity of collecting data from various fields, e.g. mental health, developmental, educational and cognitive psychology as well as epidemiological and evaluation studies. All gathered information will permit the formulation of more operational programme goals, which will lead the authors to choose prevention topics and methods. It is also easy to convert the operational goals to evaluation questions related to the key elements of the programme. In turn, evaluation questions allow the creation of evaluation indicators. The following step is the selection of measurement methods and evaluation strategy. The process description is illustrated by the example of primary prevention alcohol programme "Program Domowych Detektywow" which is the Polish adaptation of the US "Slick Tracy Home Team" programme (a part of the "Northland Project"). The problems and difficulties related to the various phases of the process of programme development are discussed. The problems with evaluation planning and finding a compromise between methodological standards and real life conditions are underlined. PMID- 11490069 TI - [Health promotion and safety promotion. New trends in injury prevention]. AB - The aim of the paper is to develop the concept of safety promotion that supports injury prevention in the last decade. The relationship between health promotion and safety promotion -was described, considering similarities, differences and possible interactions. Historically both concepts represent different traditions of public health and preventive medicine. Now they converge to a common point where actions aimed at both individuals and their life style and the community should be considered. In the second part, some international efforts to exchange experiences and provide information on effective strategies were presented. Systematic literature reviews and meta-analyses could help health care providers in setting priorities for future research and for implementations of interventions. PMID- 11490070 TI - [Monitoring the quality of obstetric care]. AB - The aim of this study was to analyse some aspects and methods use to evaluate the quality of obstetric care, and their implementation into obstetric practice. Assurance of proper quality of services in obstetrics becomes today a key problem in Poland. The article describes the evaluation of structures, processes, and results as methods of quality assurance and control. Authors suggest that special attention should by paid to correct leader- ship, medical records analysis and later formulating constructive conclusions and quality improvement. Audit and quality improvement at the national level seems to be very important. Monitoring of quality is not only necessary, but also continuous management and improvement. It is also necessary to remember that the process of quality improvement once begun should be continued. PMID- 11490071 TI - Lower limb muscle activity in ambulatory children with cerebral palsy before and after the treatment with Botulinum toxin A. AB - Purpose: The study investigated the effect of Botulinum toxin A on the gait and lower limb muscle activity of ambulatory CP children. Methods: 19 spastic diplegic and 4 left hemiparetic CP children were injected with a mean dose of 23.5 units of Botulinum toxin A/kg body weight into the gastrocnemius and hamstring muscles. Muscle tone and gait analysis including the kinesiological electromyogram of the shank and thigh muscles were assessed before and four weeks after injection and compared with the help of a multivariate analysis (p < 0.05). Results: Botulinum toxin A caused a definite reduction of plantarflexor, knee and hip hypertonia in 21 children, resulting in a more plantar grade and erect gait in 17 children four weeks after injection. Gait analysis showed a statistically significant improvement in peak ankle dorsi-flexion and knee extension during stance, and the length of the force point of action under both feet increased. Electromyography revealed sig-nificantly less co-contraction of the lower leg muscles, due to a more phasic instead of a tonic activity of the tibialis anterior muscle, and an improved activation pattern of the left rectus and biceps femoris muscles. Conclusions: The present study demonstrated that the injection of Botulinum toxin A resulted in a more mature muscle activation pattern of CP children. Most of the children walked more plantigrade and erect, the functional gait parameters, however, did not change. PMID- 11490072 TI - A synthesized pyrimidine compound, MS-818, promotes walking function recovery from crush injury of the sciatic nerve through its indirect stimulation of Schwann cells. AB - Purpose: We evaluated the effects of the drug MS-818 (2-piperadino-6-methyl-5-oxo 5,6-dihydro-(7H) pyrrolo-[3,4-d] pyrimidine maleate), a synthesized pyrimidine compound, on regeneration in crush-injured sciatic nerves of rats. Methods: MS 818 at 1.0 or 10 mg/kg or the vehicle was intraperitoneally injected into rats daity. The pinch test (PT) was performed 5 days after the operation. Walking function recovery was assessed by the sciatic nerve functional index (SFI). Time dependent changes in the levels of transcripts of nerve growth factor (NGF) and apolipoprotein E (ApoE) genes were monitored by RT-PCR. NGF peptide levels retained in the crushed nerves of rats 5 days after surgery and in the culture medium of IMS32 cells, a mouse Schwann cell line, incubated for 24 h with high or low doses of MS-818, were measured by enzyme immunoassay. Results: The PT showed that MS-818 injection promoted axonal elongation by 19.3 % compared to the vehicle injected control (n = 7, *p < 0.03). The SFIs 3 weeks after injection of MS-818 at 1.0 mg/kg and 1 0 mg/kg were significantly increased to the control level (n = 5-6, **p < 0.006 and *p < 0.03, respectively). Injection of MS-818 at 1.0 mg/kg induced NGF gene expression more than twofold compared to that of the control at 5 to 6 days after surgery (n = 4). NGF levels in crushed nerves after MS-818 injection at 1.0 and 10 mg/kg tended to be higher than those of the vehicle-injected controls by approximately 20 %, although it did not reach statistical significance. Treatment of IMS32 cells with MS-818 failed to give rise to NGF overproduction and its secretion into the culture medium. Conclusions: These present evidences suggest that MS-818 enhances functional recovery of damaged sciatic nerves by promoting axonal sprouting through indirect activation of Schwann cells and that local production of NGF rnay be activated by MS-818. PMID- 11490073 TI - Uncoupling of human short and long latency stretch reflex responses with operant conditioning. AB - Purpose: Successful operant conditioning of the biceps brachii spinal stretch reflex (SSR) has resulted in concurrent changes in the magnitude of long latency reflex responses (LLRRs). This finding suggests a coupling of the SSR and LLRR. The purpose of the present study was to downtrain the LLRR using operant conditioning and to observe any concurrent change in the SSR. Methods: Fourteen, able-bodied, human subjects were randomly assigned to either the control group or the training group. The LLRR and SSR responses were measured as magnitude of electromyographic response to a quick stretch of the elbow flexors, delivered by a torque motor. All the subjects attended fourteen sessions. The first six sessions were baseline sessions during which no conditioning or feedback occurred. The next eight sessions were the same as the baseline sessions (extended baseline sessions) for the control group; no feedback or operant conditioning of the LLRR occurred. The next eight sessions for the training group comprised the operant conditioning. Results: Operant conditioning of the LLRR resulted in a statistically significant reduction of that response within the training group and between the two groups. Also, operant conditioning of the biceps brachii LLRR did not result in concurrent changes in the magnitude of the SSR suggesting an uncoupling of these responses. Conclusions: The LLRR of the biceps brachii could be operantly conditioned without significant changes in the SSR which suggests that these two responses can be volitionally uncoupled. PMID- 11490074 TI - Axonal regeneration of mouse retinal ganglion cells by peripheral nerve transplantation; a quantitative study. AB - Purpose: Retinal ganglion cells (RGCs) of adult mammals can regenerate their axons along a segment of the peripheral nerve (PN) that is transplanted to the cut optic nerve. There have been many trials of PN transplantation to induce axonal regeneration of RGCs in adult rodents, cats and ferrets. However, because of the technical difficulty in transplant operation, PN transplantation in adult mice has not been carried out in spite of the availability of many kinds of gene manipulated animals. Here we report the procedures for successful PN transplantation in this species. Methods: We made intraretinal (IR) and retrobulbar (RB) approaches for PN transplantation. Four weeks after PN transplantation, RGCs with regenerated axons were identified by retrograde labeling with rhodamine or horseradish peroxidase applied into the PN segment. Results: A quantitative survey showed that the mean regeneration ratio was 1.0 % (n = 8) in IR transplantation, whereas it was only 0.1 % in RB transplantation (n = 11). As previously shown in other species, the regenerated RGCs were predominantly larger-bodied cells in com-parison to intact cells. Conclusion: Possible reasons for the difference in regeneration ratio between the two transplant approaches and the feature of soma size of regenerated RGCs are discussed. PMID- 11490075 TI - Single administration of GDNF into the striatum induced protection and repair of the nigrostriatal dopaminergic system in the intrastriatal 6-hydroxydopamine injection model of hemiparkinsonism. AB - Purpose: Neurotrophic factor delivery into the brain is a promising approach in the treatment of Parkinson's disease. Glial cell line-derived neurotrophic factor (GDNF) is one of the most potent neurotrophic factors for dopaminergic neurons. Although multiple injections of GDNF into the brain are commonly performed in experimental studies, the present study investigates the efficacy of using a single injection of GDNF, which may be useful in elinically applying this treatment. Methods: Unilateral 6-hydroxydoparnine (6-OHDA) administration into the striatum was perforrned in Sprague-Dawley rats to create a partial lesion of the nigrostriatal DA system. These parkinsonian model rats received a single injection of human recombinant GDNF into the same portion of the striatum either 24 h before or 4 weeks after 6-OHDA treatrnent. Results: GDNF injected into the striatum before 6-OHDA administration potently protected the dopaminergic system, as shown by the numbers of mesencephalic dopaminergie neuron cell bodies and dopaminergic nerve terminal densities in the striatum. Dopaminergic neuron cell bodies and fiber densities were also significantly restored when GDNF was given after 6-OHDA administration, although the degree of restoration was lower than in the protective experiment. ODNF administration ameliorated apomorphine-induced rotational behavior in animals receiving it either before or after 6-OHDA treatment. However, the degree of improvement was less prominent when GDNF was iniected after 6-OHDA. Conclusion: Intracerebral GDNF adininistration exerts both protective and regenerative effects on the nigrostriatal dopaminergic system, a finding which may have implications for the development of new treatment strategies for Parkinson's disease. PMID- 11490076 TI - Acceleration of peripheral nerve regeneration following FK506 administration. AB - Purpose: The severe functional and sensory deficits seen following injury to peripheral nerves makes facilitation of nerve regeneration a primary goal of the reconstructive surgeon. This study examines whether daily administration of FK506 or Cyclosporin A expedites peripheral nerve regeneration following neurotmetic injury in a rat model Methods: Inbred Buffalo rats were randomized to three experimental groups. Group I rats served as untreated controls. Rats in groups II and III received daily subcutaneous CsA (5 mg/kg), and FK506 (1 mg/kg), respectively. Each animal underwent unilateral posterior tibial nerve transection with immediate epineurial reapproximation. Functional recovery of the injured limb was assessed by serial walking track analysis. Nerve regeneration was assessed histomorphometrically via light microscopy. Results: Return of hindlimb function in control animals occurred at 32 days post injury. CsA and FK506 treated transection animals recovered at 26 and 18 days post injury, respectively. Statistically significant greater fiber density and percent neural tissue were seen in FK506- treated animals compared to control animals four weeks post transection. Conclusions: This data suggest that the daily systemic administration of both CsA and FK506 accelerate the rate of functional regeneration, following neurotmetic injuries in tbc rat model. FK506's effect on nerve growth is significantly greater than that of CsA. PMID- 11490077 TI - Phosphonate quinoxalinedione AMPA antagonists. AB - In the Western world, over 350,000 deaths and $30 billion in medical costs are attributed annually to stroke. Head and spinal cord trauma cause an estimated 250,000 deaths annually and result in medical costs of $15 billion. Although stroke and head/spinal cord trauma are leading causes of disability and death in humans, no adequate neuroprotective treatment is available. Glutamate antagonists derived from the quinoxa-linedione scaffold are as drug candidates for neuroprotection in stroke and trauma. Quinoxalinedione derivatives such as 2,3 dihydroxy-6- nitro-7-sulfamoylbenzo(f)quinoxaline and 6-(1H-imidazol-1-yl)-7 nitro-2,3-(1H,4H)-quinoxalinedione failed clinical trials because of insolu bility and resulting nephrotoxicity. Introduction of a phosphonate group into the quinoxalinedione skeleton improves solubility and leaves potency for the alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor unchanged. Phosphonate quinoxalinedione derivatives ZK202000 and ZK200775 protected rodent brain against sequelae of permanent occlusion of the middle cerebral artery and head trauma. No major deleterious effects on motor coordination, cardiovascular, or respiratory systems were detected in doses required for neuroprotection. No psychotomimetic and no neurotoxic side effects, typical for N-methyl-D-aspartate antagonists, were observed following treatment with phosphonate quinoxalinediones. PMID- 11490078 TI - Is there an optimal age for recovery from motor cortex lesions? II. behavioural and anatomical consequences of unilateral motor cortex lesions in perinatal, infant, and adult rats. AB - Purpose: The purpose of this study was to compare the behavioural and anatomical effects of unilateral motor cortex ablation in neonatal, infant, and adult rats. Methods: Rats were given unilateral lesions of the motor cortex on the day of birth (P1), at ten days of age (P10), or in adulthood. They were trained on several motor tasks (skilled forelimb reaching, beam traversing, tongue extension), general motor activity, and a test of spatial learning (Morris water task). Results: Although all lesion groups were equally impaired at skilled reaching with the forelimb contralateral to the lesion, rats with P1 lesions also were impaired at traversing a narrow beam and at learning the Morris task. Gross anatomical analyses revealed that the P1 rats had smaller brains than the other groups, a result that may account for the larger behavioural deficits in the P1 group. Analysis of Golgi-Cox stained neurons showed that relative to control groups, all lesion groups showed an increase in dendritic length in the basilar dendrites of layer III pyramidal cells and, paradoxically a decrease in length of the apical dendrites of the same cells. Conclusions: The bilateral alterations in dendritic organization following the motor cortex lesions suggest that there has been a bilateral reor-ganization of intrinsic cortical connectivity following motor cortex lesions at any age. These alterations in connectivity are likely not identical in the young and adult animals, however, because relative to controls, both the young operated groups, but not the adult group, showed a bilat-eral drop in spine density in the basilar dendrites of layer V pyramidal cells. These findings are discussed with respect to the idea that there may be critical ages in development in which animals can use anatomical modifications to compensate for deficits produced by cortical injury. PMID- 11490079 TI - Neuronal nitric oxide synthase inhibitor has a neuroprotective effect in a rat model of brain injury. AB - Purpose: The aim of the present study was to assess the effects of neuronal nitric oxide synthase (NOS I) inhibitors and a combination of NOS I and NOS II inhibitors on lesion volume after experimental brain injury. Methods: Cold lesion of the brain was induced by application of a precooled (-78 degrees C) copper cylinder to the intact dura of the rat for 6 s. Brains were removed 24 h after the injury and lesion volume determined using the triphenyltetrazolium-chloride method. Results: The specific NOS I inhibitor 3-bromo-7-nitroindazole (Br-7-NI) reduced lesion volume significantly by 21 % compared with the vehicle control. In contrast, 7-nitroindazole had no effect on lesion volume. When aminoguanidine, a specific NOS II inhibitor, was adminis-tered after Br-7-NI, lesion volume was significantly reduced but not significantly more than with either compound alone. Conclusion: Brain injury after cold lesion is partly mediated by NOS I activity and can be attenuated successfully with Br-7-NI, while coin-hibition of NOS II does not improve the outcome significantly. PMID- 11490080 TI - Training ameliorates deficits in visual detection and orienting following lesions of primary visual cortex sustained in adulthood and in infancy. AB - Purpose: Damage of primary visual cortex in adult humans, monkeys and cats severely disrupts vision by disconnecting much of the cogni-tive processing machinery of extrastriate cortex from its source of visual signals in the retina. Equivalent lesions sustained early in life result in partial sparing of visual processing functions and evidence from the mature brain suggests that systematic training procedures can ameliorate the impact of remaining deficits. The purpose of the present work was to use two reflex-based, visual detection and orienting tasks to test for the therapeutic effects of reha-bilitative training in cats that sustained lesions of primary cortical areas 17 & 18 in adulthood, and to test whether similar training is of benefit to cats that incurred equivalent lesions at one month-of-age (P28) or shortly after birth (P1). Methods: Cats were trained to attend to static visual and auditory cynosures and tested on their ability to disengage the cynosure and orient towards a target presented in the periphery of the testing arena. Targets were: 1) a high contrast, dark, moved rod; 2) an illuminated static light-emitting diode (LED); and 3) a broad band, white noise, sound stimulus. Results: On Task 1, cats with lesions of areas 17 & 18 sustained in adulthood are markedly impaired whereas cats that sustained lesions in infancy exhibit partial sparing of the visual operations underlying this task. With training, the performance of all cats improved. On Task 2, performance by the adult-lesion and the P1-groups were markedly impaired, whereas the P28-group exhibited partial sparing. On Task 3, per-formance by all groups was uniformly high. No detectable benefits of training were identified on tasks 2 and 3. Conclusions: Overall, the results show that cats incur definite benefits of training following visual cortex lesions regardless of whether the lesions were sustained early in life or later, but the benefits are limited to specific types of visual stimuli. PMID- 11490081 TI - Stroke-blind for colors, faces and locations: Partial recovery after three years. AB - Purpose. To study and follow-up achromatopsia, prosopagnosia, and topographagnosia in a patient who suffered a bilateral stroke of the posterior cerebral arteries. Methods. Ophthalmological, neuropsychological and neuroradiological examinations were conducted over a span of 3 years to assess the amount of brain damage and look for signs of functional recovery. Results. After the onset of achromatopsia, perception of the color green re-appeared first, followed by red, yellow, and brown. Blue which had appeared entirely black was last to return. While reading and color naming have largely recovered, color discrimination after three years remains poor especially in dim lighting. Similarly, with prosopagnosia, while the patient has learn to identify people (including photographs) by individual features, his ability to perceive and recognize faces and facial expressions holistically remains severely impaired. Recognition of streets, houses, and topographical layouts also continues to be affected, while the perception of speed and distance has somewhat improved. Perimetry further suggests a mild improvement of the superior hemianopia in his visual fields. Conclusions. Although major deficits in color, face and place perception remain, some functions impaired by the stroke have returned par alleling a partial recovery from the brain lesion as demonstrated by NMR. PMID- 11490082 TI - Long-term TGFbeta2 protein expression in heterotopic cortical grafts located in the rat striatum. AB - Purpose: Transforming growth factor beta 2 (TGFbeta2) is a multifunctional cytokine thought to play a crucial role in neuronal growth, differ-entiation and survival. In the cortex of adult rats, TGFbeta2 is constitutively expressed in a subset of neurons and astrocytes. In the present study we analyzed whether TGFbeta2 is also present in intrastriatal transplants of cortical anlage. In addition we investigated the temporo-spatial expression pattern of TGFbeta2 in the surrounding host striatum. Methods: Cortical primordia of rat fetuses (E14) were stereotactically grafted into the rostral striatum of adult recipient rats. Grafts were allowed to differentiate for 1, 2, 3, 4, 8 weeks or one year, respectively, followed by morphological and immunohistochemical analysis. Results: From week 2 on, TGFbeta2-immunoreactivity (IR) was detectable in transplanted neurons. Within the graft, GFAP-IR was already present one week after transplantation, whereas TGFbeta2-immunostained astrocytes were first seen after 2 weeks. One year after transplanta-tion, TGFbeta2 positive neurons and astrocytes were still present. In the host striatum and at the graft-host interface an increase of TGFbeta2-immu-noreactive astrocytes first occured after one week both in grafted animals and in sham-operated (lesioning without grafting) rats. Conclusions: Our data suggest that at least a subpopulation of transplanted neurons develops a phenotype as cortical neurons in situ with respect to TGFbeta2 expression. Upregulation of astrocytic TGFbeta2 expression within the graft, however, is due to the trauma imposed by the transplantation procedure rather than an intrinsic differentiation program of co-grafted astrocytes. PMID- 11490083 TI - Target-dependent axonal sprouting following vagal-hypoglossal nerve anastomosis in cats. AB - Purpose: To investigate the relationships between the axonal sprouting and target neurotization by central neurons after nerve heterocon-nection. Methods: Unilateral (right) vagal-hypoglossal nerve anastomosis (VHA) was performed in adult cats. Following 3-315 days postoperation (dpo), quantitative analyses and ultrastructural changes in the proximal portion of the vagal-hypoglossal heteroconnected nerve as well as the time course of neuronal regeneration were studied. Along with this, horseradish peroxidase (HRP) retrograde tracing technique was used to label the neurons of dorsal motor vagal nucleus (DMV) and nucleus ambiguus (NA) to ascertain if target neurotization was established. Results: The contralateral (left) intact vagus nerve proximal to the level of ansa cervicalis showed an average of 33 +/- 1 myelinated and 74 +/- 4 unmyelinated axons in 727 &mgr;m(2) sectional area of the nerve. In the heteroconnected nerve at the corresponding level just proximal to the anastomosis site, there was a marked increase in the number of small axons sprouting from the unmyelinated nerve fibers between 18 and 25 dpo. The number of these axonal sprouts appeared to decline at 32 dpo but its increase of 131 % was sustained until the late regeneration stage at 315 dpo when compared with the contralateral nerve serving as a control. The mean number of myelinated axons per area unit (727 &mgr;m(2)) was reduced to 18 at 3 dpo but was immediately restored to the normal range at 7 dpo. The retrograde labelling of neurons in both the DMV and NA was first detected at 22 dpo and was progressively increased peaking by about 67 dpo. Conclusions: We conclude that compared with the unmyelinated axons, the myelinated axons may acquire a superior interaction with the new target. Furthermore, the postoperative neurotization of tongue muscles may initiate and facilitate the retraction of the redundant axonal sprouts. PMID- 11490084 TI - Changes in BDNF and neurotrophin receptor expression in degenerating and regenerating rat retinal ganglion cells. AB - Purpose: Exogenously applied BDNF has been shown to rescue rat retinal ganglion cells (RGCs) from axotomy-induced apoptotic death, presumably via activation of its high affinity receptor TrkB. Since both TrkB and BDNF are endogenously expressed in RGCs, auto- or para-crine neurotrophic loops in the retina may be involved. In the present study, we investigated whether expression levels of BDNF, TrkA, TrkB, TrkC and p75 protein in RGCs are specifically regulated following axonal lesion and during regeneration of optic fibres in the adult rat. Methods: By double labelling retinal cryosections with Fluorogold and respective antibodies we determined the percentage of RGCs expressing the above-mentioned markers. In addition, mRNA levels of BDNF and TrkB were measured using quantitative RT-PCR. Results: Compared to controls the number of BDNF-positive RGCs increased twofold 2 days after axotomy and the percentage of RGCs expressing TrkB was elevated by 50 %. Correspondingly, mRNA levels of BDNF increased about twofold 2 days after axotomy. During regen-eration, the percentage of BDNF immunoreactive RGCs was further elevated compared to axotomy alone. The number of TrkA-positive RGCs doubled after axotomy, whereas no significant change in TrkC expression was observed. P75 expression was not detected in adult rat RGCs. Conclusion: Our results suggest that intrinsic rescue mechanisms may contribute to short term neuronal survival and axonal regeneration of RGCs after axonal lesions. PMID- 11490085 TI - Diethyldithiocarbamate (DEDC) impairs neuronal recovery following sciatic nerve injury in rats. AB - Purpose: Diethyldithiocarbamate (DEDC) is a substituted dithiocarbamate that is metabolically interconvertible with disulfiram (Ant-abuse). In recent years DEDC has received considerable attention because of its clinical applications and potential role in mediating both the toxic and therapeutic actions of disulfiram which is frequently used for alcohol aversion therapy. DEDC is known for its multiplicity of action that exerts both pro- and antioxidant effects. In rodents DEDC has been shown to produce neuroprotective as well as neurotoxic effects. The purpose of this study was to examine the effect of DEDC on neurological recovery following sciatic nerve crush injury (SNCI) in rats. Methods: Adult female Wistar rats were subjected to SNCI with a haemostat under deep anaesthesia. The animals were orally treated with DEDC at the doses of 250 mg/kg, 500 mg/kg and 750 mg/kg body weight 1 hr before SNCI and then once daily for 60 days. The animals were observed for sciatic functional index (walking deficit), electrophysiological and histological changes. Vitamin E level was measured to deter-mine antioxidant status of sciatic nerve. Results: Crush injury to the sciatic nerve resulted in a significant impairment of functional response which gradually recovered over a period of 22 days. Treatment of animals with DEDC caused a significant delay in functional recovery which was accompanied by poor histo-logical and electrophysiological outcome. Prooxidant effect of DEDC is quite evident from a significant decrease in vitamin E levels in both injured and uninjured sciatic nerves. Conclusions: Our results demonstrate that exposure to DEDC adversely affects recovery from peripheral nerve injury. The delay may to some extent be attributed to DEDC induced oxidative stress. PMID- 11490086 TI - Conditionally immortal neuroepithelial stem cell grafts restore spatial learning in rats with lesions at the source of cholinergic forebrain projections cholinergic forebrain projections Conditionally immortal neuroepithelial stem cell grafts restore spatial leaming in rats with lesions at the source of cholinergic forebrain projections. AB - Purpose: Loss of cholinergic projections from the basal forebrain (BF) to the cortex and from the medial septal area (MSA) to tbe hippocampus is a reliable correlate of cognitive deficits in aging and Alzheimer's disease (AD). We assessed the capacity of grafts of the conditionally immortal MHP36 clonal stem cell line to improve spatial learning in rats showing profound deficits after lesions to these projections. Methods: Rats were lesioned by infusions of S-AMPA unilaterally into BF or bilaterally into both BF and MSA. MHP36 cells were implanted ipsilaterally in cortex or basal forebrain two weeks after unilateral BF lesions, and in cortex and hippocampus bilaterally six months after bilateral BF-MSA lesions. Intact and lesion-only controls received vehicle. Six weeks later rats were assessed in spatial learning and memory tasks in the water maze, and then perfused for identification of grafted cells by beta-galactosidase immunohistocheniistry. Results: Lesioned rats with MHP36 grafts, whether implanted two weeks or six months after lesioning, learned to find a submerged platform in the water maze as rapidly as intact controls, and showed a strong preference for the platform quadrant on probe trials, whereas lesioned controls were impaired in all measures. Grafted cells of both neuronal and glial morphologies, migrated away from cortical implantation sites in BF Lesioned rats to the striatum, thalamus and basal forebrain lesion area. Cells implanted in basal forebrain showed a similar distribution. In rats with bilateral BF-MSA lesions, grafts implanted in the hippocampus migrated widely through all layers but cortical grafts largely escaped up the needle tract into the meninges. Conclusions: Although MHP36 grafts were functionally effective in both lesion models, the site and age of lesions and site of implantation influenced the pattern of engraftment. This flexibility encourages the development of conditionally immortal human stem cell lines with similar capacities for functional repair of variable neuronal degeneration in AD or aging. PMID- 11490087 TI - Methylprednisolone exacerbates axonal loss following optic nerve trauma in rats. AB - Purpose: This study investigates the clinical dogma that very high doses of methylprodnisolone helpful in spinal cord injury are also helpful in optic nerve trauma. Methods: The right optic nerve of 29 male rats received a 5 second traumatic crush followed 30 minutes later by one of five intravenous treatments (methylprednisolone 30 mg/kg, 60 mg/kg, 90 mg/kg, 120 mg/kg, or saline). Treatment was continued for three additional administrations at 6 hour intervals. Untreated sham controls (n=7) were also prepared. Six weeks after injury, animals were sacrificed, perfused and optic nerves systematically counted. Results: Axon counts (means +/- s.e.m.) were as follows: Saline = 16,670+/-8,900 (n=5); Methylprednisolone: 30 mg/kg = 8,098+/-4,741 (n=5); 60 mg/kg=6,925+/-6,517 (n=4); 90 mg/kg=2,663+/-2,653 (n=4); 120 mg/kg=6,149+/-3,487 (n=6). Consequently, the data revealed that saline treated animals retained more axons than those that were administered methylprednisolone (p<0.02). Conclusions: We conclude that methylprednisolone exacerbates axonal loss following crush injury in the rat optic nerve. Based on the results of this study, clinical studies of traumatic optic neuropathy in the future should also examine the possibility that corticosteroid treatment may have an adverse effect on visual outcome following optic nerve trauma. PMID- 11490088 TI - Axonal outgrowth in muscle grafts made acellular by chemical extraction. AB - Purpose: To compare nerve regeneration in autologous detergent extracted and freeze-thawed muscle grafts and to electrophoretically characterize the grafts. Methods: Autologous acellular muscie grafts were created either by freeze/thawing or by detergent extraction and then used to bridge a 10 mm gap in rat sciatic nerve. The autologous grafts were compared with respect to protein content, using electrophoresis preimplantation, and axonal outgrowth, Schwann cell and macrophage content, using immunocytochemistry (neurofilaments, S-100 protein, ED 1 macrophages) at 5-20 days postimplantation. Results: The extracted muscle grafts were elastic, but the amount of several proteins was reduced and laminin was still present at a position of basal laminae of the muscle fibers. The freeze/thawed grafts were brittle and lacked elasticity, but resulted in minor changes in major proteins. The axons regenerated through both types of grafts (initial delay 6 days and rate 0.7-0.8 mm/day), which shrunk in length by 25%. There were no apparent differences with respect to Schwann cells and macrophages. Conclusions: The results suggest that detergent extracted muscle tissue, in which some basal lamina proteins remain but cells are removed, could present a new favourable option for nerve grafting. PMID- 11490089 TI - Motor reorganization after spinal cord injury: evidence of adaptive changes in remote muscles. AB - Purpose: Given that SCI leads to substantial changes in biomechanical properties of the body and to widespread postlesional reorganization of the motor system as determined by functional imaging studies, we sought to identify neurophysiological correlations and time course of reorganization affecting muscles more distant to a SCI. Methods: Two arm muscles distant to a SCI (T2-L3), M.biceps brachii (BIC), M.abductor pollicis brevis (APB), were studied in 13 SCI patients and 15 controls. Motor thresholds at rest (MT), facilitatory effects on MEP-amplitudes (FE) with voluntary activation, MEP-amplitudes with maximal stimulation (MA) and recruitment curves (RC) were measured and correlated with level, age and severity of the lesion. Follow-up studies (t2) were performed in five patients with clinical recovery. Results: Patients exhibited smaller MA from activated BIC, a tendency towards smaller FE and smaller RC-slopes at t1. With clinical recovery, activated BIC-FE, MA and RC-slopes tended to normalize. Conclusions: Our data support the hypothesis that postlesional reorganization of the motor system also involves remote muscles. Considering pattern and time course of reorganization, we speculate that they appear as sequelae of the trauma, possibly representing an adaptation of the motor system to an altered biomechanical status after SCI. PMID- 11490090 TI - The Effect of Methylprednisolone on Caspase-3 Activation after Rat Spinal Cord Transection. AB - Purpose: Caspase-3 is known as a crucial effector for apoptotic cell death. Apoptosis has recently been recognized as an important cell death mechanism after spinal cord injury (SCI). This study attempts to define the effect of methylprednisolone (MP) on the activation of caspase-3 in the lesioned area following SCI. Methods: Forty-eight rats with a complete transection of the thoracic spinal cord received a placebo or MP (30 mg/kg, iv.) at 5 min, 2 and 4 h post-injury and were then sacrificed at 12, 24 h, 3 or 7 days thereafter. Results: Caspase-3 positive cells in the lesioned area were immunocytochemically observed in both cord stumps and decreased in number with increasing distance from the lesion site. More caspase-3 positive cells were present in the MP treated group than the control group at all time points, but the differences were not statistically significant. Conclusion: These results suggest that the MP induced decrease of tissue loss following SCI may not involve a reduction of apoptotic cell death. PMID- 11490091 TI - Effects-of fluoxetine on sensorimotor and spatial learning deficits following focal cerebral ischemia in rats. AB - Purpose: The present study investigated the effects of fluoxetine, a serotonin reuptake blocker, on behavioral deficits of rats subjected to transient focal cerebral ischemia. Methods: The right middle cerebral artery of rats was occluded for 120 min using the intraluminal filament method. Fluoxetine treatment (5 mg/kg, i.p.) was started 2 days after ischemia induction and treatment was continued for 10 days thereafter. Sensorimotor recovery was assessed using the limb-placing test and cognitive impairment was assessed using a water-maze test at the end of the experiment. Results: Fluoxetine treatment did not improve performance of ischemic rats in the limb-placing test. Nor was the ischemia induced deficit in the water-maze test affected by fluoxetine. The infarct volumes in the cortex or striatum, determined after the experiment, were not different between ischemic groups. Conclusion: These results suggest that subchronic fluoxetine treatment following experimental focal cerebral ischemia is not detrimental to behavioral outcome, but it also does not enhance spontaneous sensorimotor recovery or attenuate spatial learning deficits. PMID- 11490092 TI - Nerve growth factor and striatal glutathione metabolism in a rat model of Huntington's disease. AB - Purpose: To test the influence of nerve growth factor (NGF) on striatal glutathione (GSH) content and the activities of GSH-related enzymes from quinolinic acid-lesioned rats. Methods: Rats were intrastriatally injected with QA and NGF. Enzymatic and GSH assays were performed one week later. Results: NGF prevented the QA-induced decline in glutathione reductase activity and GSH content. Conclusions: NGF is able to prevent some of the disturbances induced by the excitotoxic insult in the striatal GSH metabolism. PMID- 11490093 TI - Fibroblast growth factor-8b-stimulated myogenic cell proliferation is suppressed by the promyelocytic leukemia gene. AB - Muscle cell growth is regulated by growth-promoting and -inhibiting factors. In this study, the physiological effects of fibroblast growth factor (FGF)-8b and the promyelocytic leukemia (PML) gene on G8 myogenic cells were examined. FGF-8b was found to strongly stimulate myogenic cell proliferation. Signal transduction assays using AP-1/SEAP and E-box/SEAP reporters revealed that the transcriptional factors junB/c-fos and c-myc were involved in FGF-8b-stimulated G8 cell growth. Besides examining factors that positively stimulate myogenic cell growth, we also examined genes that negatively affect cell growth. PML is a growth suppressor gene and we studied its expression in G8 cells under different growth conditions. Immunohistochemical staining revealed that in the presence of low serum, PML was expressed in approximately 23.2% of all cultured G8 cells. However, under normal culture conditions (10% serum), PML expression dropped to about 2.6%. We found that the PML gene acted antagonistically to FGF-8b, as the overexpression of PML in G8 cells significantly inhibited FGF-8b-stimulated cell proliferation. It also inhibited AP-1 and E-box transactivation. However, we believe that PML functions as a stress-response gene in G8 cells rather than as a gene normally involved in regulating muscle development. PMID- 11490094 TI - The anti-tumour effect of Klebsiella pneumoniae capsular polysaccharides. AB - K24 capsular polysaccharide (K24-CPS), with a known structure of a repeating unit, was isolated from the capsule of Klebsiella pneumoniae serotype K24. The polysaccharide was found to suppress the proliferation of Ehrlich ascites tumour (EAT) cells in vitro, but did not alter the cell cycle distribution of cells. K24 CPS treatment reduced the tyrosine phosphorylation of some proteins in EAT cells. Furthermore, the treatment also decreased the expression of c-JUN, but had no effect on the levels of c-FOS and c-MYC. It is speculated that the growth suppression effect of K24-CPS may be related to its effect in down-regulating c JUN expression. PMID- 11490095 TI - The responses of rat liver glucocorticoid receptors and genes for tyrosine aminotransferase, alpha-2-macroglobulin and gamma-fibrinogen to adrenalectomy-, dexamethasone- and inflammation-induced changes in the levels of glucocorticoids and proinflammatory cytokines. AB - The responses of liver glucocorticoid receptor (GR) and genes coding for a glucocorticoid-inducible tyrosine aminotransferase (TAT) and two acute-phase proteins (APP) [alpha2-macroglobulin (alpha2-M) and gamma-fibrinogen (Fb)] to changes in glucocorticoid (GC) and proinflammatory (AP) cytokine contents have been examined in rats after single or combined treatments with turpentine oil, dexamethasone (Dex) and adrenalectomy. Activation of two APP genes in turpentine induced inflammation was accompanied by an increase in the level of GR mRNA and a preferential translocation of GR-GC complexes to the nucleoplasm, while the expression of TAT remained unaltered. Dex alone caused a decrease in the levels of GR and Fb mRNAs, activation of TAT and alpha2-M genes, a decrease in the affinity of hormone binding sites and redistribution of translocated GR-Dex complexes within the nuclei. Inflammation potentiated the effect which Dex alone exerted on the GR content and the number of GR binding sites but counteracted its influence on the affinity of GR binding sites and nuclear distribution of GR-Dex complexes. Adrenalectomy promoted a fall in TAT mRNA, no changes in the GR and Fb mRNA, a decrease in the affinity of GR hormone binding sites and redistribution of GR-hormone complexes within the nuclei. The AP cytokines released in response to inflammation exerted a counteracting effect on the adrenalectomy-induced changes in the affinity of hormone binding sites and nuclear distribution of GR hormone complexes. They potentiated a fall of TAT mRNA but promoted full expression of the Fb gene. These results argue strongly for the influence of AP cytokines on the functional state of the GR and GC signaling pathways. PMID- 11490096 TI - Effects of halothane, pentobarbital and ketamine on serum melatonin levels in the early scotophase in New Zealand white rabbits. AB - In mammals, the nocturnal rise in pineal melatonin is regulated by signals from the endogenous clock, the hypothalamic suprachiasmatic nuclei. There have been few reports on whether anaesthetics which modulate multisynaptic neuronal functions affect melatonin secretion. We studied the effects of three commonly used anaesthetics, halothane, pentobarbital and ketamine, on serum melatonin levels in male New Zealand white rabbits. Seven blood samples were collected, 30 60 min apart, before, during and after anaesthesia. Experiments were performed in the late light and early dark period, so that changes in melatonin secretion would be reflected in the onset and/or level of nocturnal serum melatonin. Serum melatonin levels were determined by radioimmunoassay. Our results indicated that halothane attenuated the release of melatonin and pentobarbital had no apparent effect, whereas ketamine potentiated the release of melatonin. These findings suggest that melatonin levels may be affected in patients anaesthetized with halothane or ketamine, resulting in disturbed biological rhythms, especially the sleep-wake cycle following recovery. PMID- 11490097 TI - Maternal transfer of melatonin alters the growth and sexual maturation of young Indian palm squirrel Funambulus pennanti. AB - To date, the phenomenon of maternal transfer of hormones to the young is an enigma. The present study explains for the first time the maternal transfer of melatonin (MEL) to the young, affecting neonatal growth and sexual maturation. The suckling pups of MEL-treated mothers exhibited significant decreases in body, testicular, vas deferens (male pups), ovarian and uterine (female pups) weights and increases in pineal gland activity along with high plasma MEL levels. The plasma level of testosterone decreased significantly in male pups, while estradiol increased and progesterone decreased in female pups of MEL-treated mothers. These results clearly suggest that MEL could be transported from the mothers to their young postnatally via the milk in order to influence neonatal growth and sexual maturation. Our results support the earlier concept and show for the first time that MEL can be transported from the mother to the young either prenatally through the placenta or postnatally via the milk. Therefore, maternal MEL can act as a biological signal for neonatal growth and sexual maturation. PMID- 11490098 TI - Tumour necrosis factor induced an early release of superoxide and a late mitochondrial membrane depolarization in L929 cells. Increase in the production of superoxide is not sufficient to mimic the action of TNF. AB - Tumour necrosis factor alpha (TNF) cytotoxicity is mediated, at least in part, by oxidative stress. One of the post-receptor events shortly after the addition of TNF is the generation of the superoxide anion (O2-*). In the present study, we attempted to examine the role of O2-* in the regulation of mitochondrial membrane potential (Delta(Psi)m) and the release of cytochrome c (cyto c) in L929 cells after stimulation with TNF. Challenge of cells with TNF (50 ng/ml) resulted in an early (30 min after the addition of TNF) increase in the production of O2-*. The use of mitochondrial electron transport chain inhibitors such as antimycin A and rotenone could, respectively, potentiate or suppress the TNF-mediated release of O2-* and cytotoxicity. TNF also induced a late (>3 h after the addition of TNF) depolarization in the Delta(Psi)m. Reduction in the release of O2-* by rotenone (50 microM) or thenoyltrifluoroacetone (250 microM) suppressed both the TNF mediated Delta(Psi)m depolarization and cyto c release. However, increase in the production of O2-* by antimycin A (25 microM) only slightly enhanced the TNF effect in altering the Delta(Psi)m and the release of cyto c. Treating cells with antimycin A alone could not induce a reduction in Delta(Psi)m nor a release of cyto c. Taken together, our results indicate that TNF induced damage in mitochondria in L929 cells. Our data also show that an increase in the production of O2-* was important in the TNF cytotoxicity, but was not sufficient to mimic the action of TNF. PMID- 11490099 TI - L-arginine immunoreactive enteric glial cells in the enteric nervous system of rat ileum. AB - L-arginine is a precursor of nitric oxide (NO) that may be involved in neuronal activity in the gastrointestinal tract. It is known that NO is formed from L arginine by NO synthase which is localized in neurons in the enteric nervous system. The present study demonstrated that significant L-arginine immunoreactivity was present in the enteric ganglia. Ultrastructural examination showed that L-arginine immunoreactivity was present in the ganglionic glial cells but not in neurons. These findings suggest that enteric glial cells may represent the main reservoir of L-arginine, which may possibly be transferred to neurons when used. PMID- 11490100 TI - Fatigue after stroke: a major but neglected issue. AB - Subjective fatigue, defined as a feeling of early exhaustion developing during mental activity, with weariness, lack of energy and aversion to effort, remains virtually unstudied in patients with stroke, bur recent surveys suggest that it is a major, commonly overlooked, stroke sequela. While the few existing series did not show significant correlations between fatigue and stroke severity, lesion location, cognitive and neurological impairment and depression, recent neurobehavioral studies have highlighted an association between fatigue and brainstem and thalamic lesions. This suggests that fatigue may be linked to the interruption of neural networks involved in tonic attention, such as the reticular activating system. In fact, several subtypes of fatigue may develop after stroke, in connection with cognitive sequelae, neurological impairment, psychological factors and sleep disorders. A challenge is to identify and delineate these different subtypes and to distinguish them from mood disorders, which frequently coexist. We emphasize the concept of 'primary' poststroke fatigue, which may develop in the absence of depression or a significant cognitive sequela, and which may be linked to attentional deficits resulting from specific damage to the reticular formation and related structures involved in the subcortical attentional network. In the patients with excellent neurological and neuropsychological recovery, poststroke fatigue may be the only persisting sequela, which may severely limit their return to previous activities. The recognition of poststroke fatigue may be critical during recovery and rehabilitation after stroke. PMID- 11490101 TI - Characterization of cerebral small vessel disease by proton spectroscopy and morphological magnetic resonance. AB - This study sought to investigate whether clinical and neuropsychological impairment in cerebral small vessel disease (CSVD) can be evaluated by proton spectroscopy ((1)H-MRS) and structural magnetic resonance (MR) imaging. Sixteen patients with CSVD and 15 healthy age-matched controls participated in the study. In addition to spectroscopic and structural MR examination all patients underwent a comprehensive clinical and neuropsychological investigation. Significant differences in between patients and controls were revealed by (1)H-MRS in the parietal white matter: decreased metabolic ratios of N-acetyl aspartate to choline (NAA/Cho; patients: 1.37 +/- 0.17, control: 1.72 +/- 0.25, p < 0.001) and of N-acetyl aspartate to creatin (NAA/Cr; patients: 1.41 +/- 0.15, control: 1.66 +/- 0.2, p < 0.01) indicated a pathological state. Evaluation of spectroscopic and neuropsychological data revealed a close relation between attentional impairment, i.e. delayed cerebral transmission time and decreased NAA/Cho and NAA/Cr (r = 0.62, p = 0.014). In sum, (1)H-MRS allowed a clear discrimination between patients with CSVD and age-matched normal controls. Moreover, comparisons of (1)H-MRS and neuropsychological data suggested that NAA metabolic levels, and particularly the delay in cerebral transmission time, could be potential predictors of the severeness of attentional impairment. PMID- 11490102 TI - Hyperhomocyst(e)inemia is a risk factor of secondary vascular events in stroke patients. AB - OBJECTIVE: Moderate hyperhomocyst(e)inemia is an independent risk factor for stroke, but it is unclear whether it also would be a risk factor for secondary vascular events after stroke. METHODS: Longitudinal study of 137 consecutive ischemic stroke patients (age 45-91 years) who were prospectively studied with a standard clinical protocol. Vascular events (stroke recurrence, ischemic heart disease, deep venous thrombosis or peripheral arterial disease) were identified during 2 years of follow-up. Serum homocyst(e)ine was determined 3 months after the stroke. The cumulative proportion of patients with homocyst(e)ine above or below the 75th percentile who survived free of vascular events was determined by Kaplan-Meier analysis. Cox models were used to estimate the relative risk of vascular events after controlling for other confounding factors. RESULTS: Serum homocyst(e)ine was significantly higher in patients with vascular events (26.2 versus 19.4 micromol/l; p = 0.016). The cumulative proportion of patients with vascular events was 46.5% in the group with homocyst(e)ine over the 75th percentile (>30 micromol/l) and 20.2% in the other group (log-rank test 7.5; p = 0.0062). After adjustment for age, sex, high blood pressure, diabetes, heart disease, previous cerebrovascular disease, smoking and serum cholesterol, the relative risk of vascular event for patients above compared with those below the 75th percentile of serum homocyst(e)ine was 2.8 (CI 95% 1.3-6; p = 0.01). CONCLUSION: Hyperhomocyst(e)inemia is a significant risk factor for vascular events after ischemic stroke. This finding is independent of other risk factors such as hypertension, and may have therapeutic relevance in the secondary prevention of vascular diseases in stroke patients. PMID- 11490103 TI - Lack of evidence for a poor haemodynamic or metabolic state of the brain in patients with haemodynamic clinical features associated with carotid artery occlusion. AB - OBJECTIVE: To assess whether patients with carotid artery occlusion (CAO) who have clinical features suggesting a haemodynamic origin have a poor haemodynamic or metabolic state of the brain. METHODS: In 117 patients with ischaemic symptoms of the eye or brain that were transient or at most moderately disabling and associated with a CAO, we compared CO(2) reactivity, quantitative flow measurement by magnetic resonance (MR) angiography, metabolic ratios measured by (1)H-MR spectroscopy, collateral blood flow patterns and the presence of infarcts of the borderzone type between patients grouped by the following clinical features: (1) presence or absence of at least one of the 'classical' haemodynamic symptoms: limb shaking, retinal claudication, precipitation of symptoms by exercise, by rising from a sitting or lying position, by transition from a cold to a warm environment, or by documented hypotension, and (2) symptoms having occurred after demonstration of the CAO or only before the occlusion was documented. RESULTS: Patients with (n = 16) and without (n = 101) one of the 'classical' haemodynamic symptoms did not differ in any of the measured indices. Patients with recurrent symptoms after documentation of the CAO (n = 56) had lower CO(2) reactivity (difference 8.3%, 95% confidence interval 0.1-16.5) than those with symptoms only before documentation of the occlusion (n = 61), whereas no significant differences were found in any of the other measured indices. The difference in CO(2) reactivity was no longer significant after adjustment for the interval between the patients' last symptoms and the CO(2) reactivity measurement. CONCLUSION: In patients with CAO we could not find an association between symptoms that have been associated with hypoperfusion and a poor haemodynamic or metabolic state of the brain. PMID- 11490104 TI - Retinal ischemia and embolism. Etiologies and outcomes based on a prospective study. AB - OBJECTIVES: To identify the most likely mechanisms of retinal ischemia and embolism in a hospital-referred population, and to determine the frequency of recurrent vascular events during the 3-month period following initial presentation. METHODS: Consecutive patients presenting to 2 tertiary medical centers and their outpatient clinics were prospectively enrolled over a 22-month period. Eligible patients presented with histories of transient or permanent monocular visual loss, or had evidence of asymptomatic retinal embolism on routine ophthalmological examination. They underwent a rapid and standardized evaluation that included imaging studies as well as blood tests, and follow-up was obtained at 1 and 3 months. RESULTS: Seventy-seven patients were enrolled. Enrollment diagnoses consisted of amaurosis fugax (n = 32), asymptomatic retinal embolism (n = 34), and central or branch retinal artery occlusion (n = 11). Eight different presumed etiologies of retinal artery distribution embolism or hypoperfusion were identified. Extracranial internal carotid artery occlusion or more than 50% stenosis was observed in 17/77 (22.1%) cases, making it the largest etiologic subgroup. Uncommon but treatable conditions were identified in 8/77 (10.4%) patients, and an etiologic diagnosis could not be made in 35/77 (45.5%) patients. Recurrent events occurred in, respectively, 14/77 (18.2%) and 6/73 (8.2%) patients at the 1- and 3-month follow-ups. They included 2 infarcts and 2 deaths; ischemic events of the retina were more common than those involving the brain. CONCLUSION: Severe stenosis of the extracranial internal carotid artery is the most common identified condition associated with retinal ischemia and embolism, but a variety of other, potentially treatable, conditions can be diagnosed if appropriate and specific evaluations are conducted. The frequency of recurrent vascular ischemic events is highest during the 1st month of follow-up and decreases during the 2nd and 3rd months. Recurrences range from relatively innocuous episodes of amaurosis fugax to vascular death. PMID- 11490105 TI - Determinants of social outcome measured by the Frenchay Activities Index at one year after stroke onset. AB - We report an investigation to identify patient factors which influence social outcome measured by the Frenchay Activities Index at 12 months after a disabling stroke. Using data from a previous randomised trial a logistic regression model identified gait speed, prestroke Frenchay Activities Index, the Abbreviated Mental Test score, sensory neglect, chronic obstructive airways disease and left hemiplegia as significantly associated with 12-month social outcome scores. The model correctly predicted the social outcome for 78% of the study population and, when tested on a second data set, correctly predicted 76% of cases. These patient factors provide insight into social activity recovery after stroke and could be considered as stratification factors in future community stroke randomised trials. PMID- 11490106 TI - Stroke pattern interpretation: the variability of hypertensive versus amyloid angiopathy hemorrhage. AB - INTRODUCTION: It is commonly felt that cerebral amyloid angiopathy (CAA) related intracerebral hemorrhage (ICH) can be distinguished from hypertension (HTN) related ICH by certain typical features on computerized tomography (CT) and magnetic resonance imaging (MRI). The purpose of this study was to investigate the performance of clinicians who were asked to differentiate between CAA and HTN based on hemorrhage pattern interpretation and to assess the feasibility of such classification. METHODS: The admission scans from 83 patients who were admitted to our service with an acute ICH were presented to 5 clinicians in a randomized and blinded fashion (1 junior, and 1 senior neurosurgical resident, 1 attending neurosurgeon, and 2 neurosurgeon-neuroradiologists). There were no patients who received oral anticoagulants other than low-dose aspirin, or who suffered from vascular malformations or tumors. Scans from 41 patients with a histologically proven diagnosis of CAA and from 42 patients with a clear history of HTN were investigated. Hematoma evacuation was done in all CAA patients and in 59% of HTN patients (n = 25). RESULTS: The overall average classification accuracy was 66.8% (range: 62.7-69.9). For correct HTN classification it was 69.5% (range: 64.3-81), and 63.9% for CAA, respectively (range: 48.9-75.6). There were negligible differences in classification accuracy among all observers. Patients with a CAA related ICH were significantly older than patients with a HTN-related ICH (74 vs. 66.5 years, p < 0.05). There was a significantly higher number of hematomas >30 ml in CAA (85.3%) when compared with HTN (59.5%). No basal ganglionic hemorrhage was seen in CAA, but in 40.5% in HTN. Intraventricular hemorrhage was seen in 24.4% in CAA, and in 26.2% in HTN. Two patients (4.9%) with CAA, and 7 patients with HTN (16.7%) presented with cerebellar hematomas. CONCLUSIONS: Three of 10 scans were not correctly diagnosed regardless of the examiner's level of training. This calls into question the reliability of classifying the underlying pathological condition based on hemorrhage pattern interpretation on CT or MRI. The definite diagnosis of CAA- versus HTN-related hemorrhage requires a histopathological confirmation and should not be based solely on hemorrhage pattern interpretation. PMID- 11490107 TI - Randomized controlled trial of rehabilitation at home after stroke: one-year follow-up of patient outcome, resource use and cost. AB - BACKGROUND AND PURPOSE: This study sought to evaluate early supported discharge and continued rehabilitation at home after stroke, at a minimum of 6 months after the intervention, in terms of patient outcome, resource use and health care cost. METHODS: Eighty-three patients, moderately impaired 5-7 days after acute stroke, were included in a randomized controlled trial, 42 being allocated to the intervention and 41 to routine rehabilitation. One-year follow-up of patient outcome included mortality, motor capacity, dysphasia, activities of daily living, social activities, perceived dysfunction, and self-reported falls. Resource use over 12 months included inpatient hospital care, outpatient health care, use of health-related services, informal care, and cost of health care. RESULTS: On univariate analysis there was no difference in patient outcome. Multivariate regression analysis showed that intervention had a significant effect on independence in activities of daily living. A significant difference in inpatient hospital care, initial and recurrent, was observed, with a mean of 18 (intervention) versus 33 days (control) (p = 0.002). Further significant differences were that the control group registered more outpatient visits to hospital occupational therapists (p = 0.02), private physical therapists (p = 0.03) and day-hospital attendance (p = <0.001), while the intervention group registered more visits to nurses in primary care (p = 0.03) and home rehabilitation (p = <0.001). Other differences in outcomes or resource utilization were nonsignificant. CONCLUSION: In Sweden, early supported discharge with continued rehabilitation at home proved no less beneficial as a rehabilitation service, and provided care and rehabilitation for 5 moderately disabled stroke patients over 12 months after stroke onset for the cost of 4 in routine rehabilitation. PMID- 11490108 TI - Ventricular fibrillation during off-pump coronary artery bypass grafting: transcranial Doppler and clinical findings. AB - A 73-year-old male developed ventricular fibrillation which lasted for 2 min and 24 s, during off-pump coronary artery bypass grafting. Cerebral hemodynamics were assessed by continuous transcranial Doppler monitoring. Ventricular fibrillation resulted in an immediate fall in cerebral blood flow velocities to almost zero with only slight fluctuations. This was then followed by a pronounced reactive hyperperfusion. Cerebral magnetic resonance imaging examinations and detailed neurological and neuropsychological evaluations were performed before and at 3 and 12 months after surgery. No evidence of cerebral damage was found. PMID- 11490109 TI - Vertebra, rib and spinal cord infarction caused by probable fibrocartilaginous embolism. PMID- 11490110 TI - Rapid resorption of intraventricular hemorrhage after thrombolytic therapy of ischemic stroke. PMID- 11490111 TI - Somatic cell exchange occurs between mouse fetal gonads and mesonephroi during in vitro culture. AB - Mesonephric cell contribution to the developing mouse gonad has previously been shown to be male-specific in in vitro culture models. This study shows that somatic cell exchange occurs between fetal mouse mesonephroi and gonads during in vitro culture of assembled tissues. Assembling gonads with cell marker-carrying mesonephroi and vice versa identified cell exchange. Two different cell markers were used. One of the markers was the thymidine analogue bromodeoxyuridine (BrdU), which was added to the culture medium during an initial culture period before in vitro culture of assembled tissues in medium without BrdU. The other was bacterial beta-galactosidase, which is expressed ubiquitously in tissues from the transgenic mouse strain ROSA26. The tissues were taken from either 10.5- or 11.5-day-old mouse fetuses, and the assembled tissues were cultured for 36-60 h. Both cell markers demonstrated the occurrence of a cell exchange in both directions between the mesonephros and the gonad. The cell contribution was found to be minor and independent of the initial age of the tissue and hours in culture. Further, mesonephric cell contribution was not male-specific. Cell exchange between the two tissues may be due to growth between the assembled tissues rather than active signal-regulated migration. It is suggested that in vitro culture models may not be applicable for investigating mesonephric cell contribution to the developing mouse gonad. PMID- 11490112 TI - Localization of cells of the prospective neural plate, heart and somites within the primitive streak and epiblast of avian embryos at intermediate primitive streak stages. AB - By constructing avian transplantation chimeras using fluorescently-labeled grafts and antibodies specific for grafted cells, we have generated a prospective fate map of the primitive streak and epiblast of the avian blastoderm at intermediate primitive-streak stages (stages 3a/3b). This high-resolution map confirms our previous study on the origin of the cardiovascular system from the primitive streak at these stages and provides new information on the epiblast origin of the neural plate, heart and somites. In addition, the origin of the rostral endoderm is now documented in more detail. The map shows that the prospective neural plate arises from the epiblast in close association with the rostral end of the primitive streak and lies within an area extending 250 microm rostral to the streak, 250 microm lateral to the streak and 125 microm caudal to the rostral border of the streak. The future floor plate of the neural tube arises within the midline just rostral to the streak, confirming our earlier study, but unlike at the late-primitive streak stages when both Hensen's node and the midline area rostral to Hensen's node contribute to the floor plate, only the area rostral to the primitive streak contributes to the floor plate at intermediate primitive streak stages. Instead of contributing to the floor plate of the neural tube, the rostral end of the primitive streak at intermediate primitive-streak stages forms the notochord as well as the rostromedial endoderm, which lies beneath the prechordal plate mesoderm and extends caudolaterally on each side toward the cardiogenic areas. The epiblast lateral to the primitive streak and caudal to the neural plate contributes to the heart and it does so in rostrocaudal sequence (i.e., rostral grafts contribute to rostral levels of the straight heart tube, whereas progressively more caudal grafts contribute to progressively more caudal levels of the straight heart tube), and individual epiblast grafts contribute cells to both the myocardium and endocardium. The prospective somites (i.e., paraxial mesoderm) lie within the epiblast just lateral to the prospective heart mesoderm. Comparing this map with that constructed at late primitive-streak stages reveals that by the late primitive-streak stages, prospective heart mesoderm has moved from the epiblast through the primitive streak and into the mesodermal mantle, and that some of the prospective somitic mesoderm has entered the primitive streak and is undergoing ingression. PMID- 11490113 TI - Morphological characteristics of dying cells in axial structures of developing human embryos. AB - Programmed cell death (PCD) is a widespread phenomenon in the development of vertebrates. In most cases, dying cells during development exhibit generalized morphological features typical of apoptosis. We analyzed the morphological features of dying cells in the developing axial structures of 5 human embryos between 5 and 8 weeks of postovulatory age. Cell death in the axial structures, i.e. spinal cord, notochord and surrounding mesenchyme and somites, was analyzed using light and electron microscopy. Tissue samples were taken from the cervicothoracic region of normal human conceptuses. Two morphological types of cell death were found: apoptosis which was characterized by round or semilunar nuclear chromatin condensations, condensation and shrinkage of the cytoplasm and formation of apoptotic bodies, and cell death without the morphological features of apoptosis which was characterized by pyknotic nuclear chromatin condensations, vacuolated cytoplasm and the formation of numerous intercellular spaces. Apoptotic death occurred during the 5th week of normal development in all the axial structures. Later, apoptotic death appeared in all the axial structures, with the exception of the notochord, where some dying cells displayed features of secondary necrosis. According to our findings, apoptosis seems to be the most frequently observed type of PCD, but it is not the exclusive type of morphological cell death during the development of axial structures in human embryos. PMID- 11490114 TI - Immunohistochemical expression of S-100 protein in human embryonal fat cells. AB - The aim of the present study was to investigate immunocytochemically the presence of S-100 protein in subcutaneous fat cells during early human embryogenesis (6-12 weeks of gestation). We found that preadipocytes in the subcutaneous tissue which were at different stages of differentiation were positive for S-100 protein. The other cells in the embryonal subcutis (mesenchymal cells differentiating into fibroblasts and fibrocytes and endothelial cells) showed a negative reaction for S-100. Our results imply that the S-100 protein is expressed from the beginning of lipidogenesis and possibly acts as a factor regulating lipid storage and body fat formation. It can be used as a reliable biochemical marker of human fat cell differentiation and for distinguishing them from the mesenchymal and the fibroblast cells in the human embryonal subcutis. PMID- 11490115 TI - Mammary epithelial cell and adipocyte co-culture in a 3-D matrix: the first step towards tissue-engineered human breast tissue. AB - Reconstruction of the female breast after cancer surgery is a demanding task where the methods used today suffer from several disadvantages. In the present study we have investigated the possibility to use tissue engineering methods to regenerate human autologous breast tissue. Human mammary epithelial cells and preadipocytes were derived from breast tissue biopsies from healthy women undergoing reduction mammoplasty, and the two celltypes were co-cultured with conventional cell culture methods as well as in 3-D matrices. The study shows that it is possible to harvest both human mammary epithelial cells and preadipocytes in a single session, propagate several subcultures, and that the cells maintain a normal intercellular distribution and growth-pattern when co cultured in a 3-D collagen gel. We propose that growth and formation of a tissue closely resembling normal human breast tissue be readily obtained in the described in vitro cell culture set-up using basic tissue engineering principles. This concept may be of great importance in the development of new methods for reconstruction of the human breast. PMID- 11490116 TI - Cryopreservation of artificial cartilage: viability and functional examination after thawing. AB - In biomedical research and in reconstructive surgery, preservation of intact tissue has been an unsolved problem. In this study, we investigated the viability of cryopreserved artificial cartilage and its synthetic activity of cartilage specific matrix proteins after thawing for in vitro use. A polymer fleece cylinder (diameter = 3 mm; height = 3 mm) was loaded with a suspension of bovine chondrocytes (25 x 10(6)/ml) and encapsulated with fibrin glue. After a culture period of 1 week, the artificial cartilage units were frozen in a cryoprotection solution containing 10% basal medium (RPMI 1640), 10% DMSO and 80% FCS. The freezing procedure consisted of three steps: a 30-min period at +4 degrees C followed by a 24-hour storage at -80 degrees C. After that, the tissue units were transferred into liquid nitrogen (-196 degrees C) for final storage. Using histochemical staining techniques of cryogenic slices, we investigated the ability of cryopreserved artificial cartilage to produce its specific matrix after thawing. A modified MTT assay was used to determine the viability of frozen tissue units in comparison with unpreserved samples at different moments after thawing. Depending on the chondrocytes used for the formation of artificial cartilage, the viability of cryopreserved tissue varied between 65 and 85%. Both the intensity of alcian blue staining for proteoglycans and the azan staining for collagens increased proportionally with incubation time after thawing. These findings indicate that cryopreservation of small artificial cartilage units is possible with a minor loss of cell viability. Secondly, its synthetic activity of cartilage-specific matrix did not decline after the freezing process. PMID- 11490117 TI - Expression of nucleotide P2X receptor subtypes during spermatogenesis in the adult rat testis. AB - The expression of ATP-gated ion channels (P2X receptors) was investigated in testes of adult rats by immunohistochemistry and Western blotting with antibodies against all seven P2X receptor subtypes. Immunoreactive cells were identified and monitored during germ cell maturation. Results of immunohistochemical and Western blotting experiments showed the expression of P2X(1), P2X(2), P2X(3), P2X(5) and P2X(7) receptors, while P2X(4) and P2X(6) receptors were absent from the testis. Blood vessels displayed immunostaining for P2X(1) and P2X(2) receptors; the P2X(1) receptors were present exclusively in blood vessels. P2X(2), P2X(3) and P2X(5) receptors were found to be expressed differentially in the various germ cell types throughout the different stages of the cycle of the seminiferous epithelium; P2X(2) and P2X(3) receptors were always observed together in the same cell types and at the same stages. Sertoli cells also showed differential staining for P2X(2) and P2X(3) receptors during the cycle of the seminiferous epithelium, whereas P2X(7) receptor expression was present throughout all stages. No immunostaining for P2X receptors was detected on Leydig cells. The possible roles of purinergic signalling in the control of germ cell maturation are discussed. In particular, it is suggested that purinergic signalling may play a role in controlling the maturation of germ cell subsets of different developmental ages that exist alongside each other in the adult testis. PMID- 11490118 TI - Muscle spindles in extraocular muscles of human infants. AB - The capacity of muscle spindles in adult human extraocular muscles (EOM) to provide effective proprioception was questioned on the grounds of their peculiar morphology. Their appearance could be attributable to ageing and to test this possibility examples of infant muscle spindles have been examined. Forty encapsulated structures from five extraocular muscles removed post mortem from 4 infant patients aged 6 days, 5, 23 and 30 months were examined by means of light microscopy using serial transverse sections. Seven of them were identified as false spindles. The remaining 33 structures, identified as spindles, contained a total of 175 intrafusal fibres varying from 2 to 12 (mean: 6) in each. 130 of these fibres (74.3%) were of nuclear chain type. Unequivocal evidence of bag fibres was not found. Spindles lacked or had a limited equatorial expansion, and the inner capsule was incomplete and irregularly shaped. 45 (25.7%) of the intrafusal fibres had extrafusal features with large diameters, peripherally placed nuclei, no equatorial modification and without associated sensory nerve terminals. Serial sections revealed that a majority of the nuclear chain fibres were interrupted, fragmented or terminated abruptly, and most spindles contained at least one incomplete fibre. These observations show that the atypical features observed in adult human EOM spindles are also present in infants and are therefore not attributable to ageing. PMID- 11490119 TI - Muscle fibre damage and regeneration resulting from surgical limb distraction. AB - Using an animal model of limb distraction, the extent of muscle fibre damage and atrophy resulting from distraction at two different rates (1.3 or 3.0 mm day(-1)) was investigated. It was found that at the high rate of distraction there was a significantly greater loss of range of joint movement and more muscle fibre atrophy and fibre damage than at the low rate. Muscle fibre damage is usually followed by regeneration. This involves the expression of the neonatal form of myosin heavy chain, which can therefore be used as an indicator of regeneration. It was found that whilst many more fibres showed evidence of damage at the high compared to the low rate, the number of fibres expressing neonatal myosin was significantly reduced, indicating the presence of a population of fibres which was undergoing degeneration without subsequent regeneration. Thus it would appear that beyond a certain rate of distraction, regeneration may be insufficient to replace contractile material damaged by overstretching. It is suggested that these fibres are replaced with connective tissue. This process may contribute to the muscle weakness and loss of range of joint movement which sometimes accompanies limb distraction procedures. PMID- 11490120 TI - In vivo measurement of fascicle length and pennation angle of the human biceps femoris muscle. AB - The purpose of this investigation was to measure in vivo fascicle length (L(fas)) and pennation angle (PA) of the long head of the biceps femoris muscle (BF(lh) m.) at different hip and knee angles while the muscle was relaxed using ultrasonography (US). Data were collected from 18 healthy females (23.0 +/- 1.8 years). To validate the measurements of the L(fas) and PA, the US measurements on cadavers (n = 5), embalmed with the hip and knee in anatomical position, were compared to actual fiber lengths and PA from the dissected muscles. US images from volunteer subjects were recorded when the hip joint was positioned at 0 degrees, 45 degrees and 90 degrees of hip flexion, and at each hip position, the knee joint was placed at 0 degrees, 45 degrees and 90 degrees from full extension. The images were digitized to measure L(fas) and PA. Results showed no significant differences between US and direct measurements of the L(fas) and PA on the cadaver BF(lh) m. (p > 0.05). L(fas) and PA changed significantly between the different hip and knee positions in volunteer subjects (p < 0.05). Changes in the L(fas) and PA are more sensitive to changes in hip position with the knee position constant than to changes in knee position with the hip position constant. This difference may be related to the larger muscle moment arm at the hip resulting in greater excursion of the muscle with changing hip position. Based on the changes in L(fas) with changing joint positions, BF(lh) sarcomere length was estimated to occupy a portion of the ascending limb, the plateau and descending limb of the length-tension relationship over the range of motion studied. US scanning is valid and reliable for measuring L(fas) and PA of the BF(lh) m. An understanding of the dynamic nature of muscle architecture will assist in determining effective and efficient clinical evaluation and rehabilitation techniques. PMID- 11490121 TI - Ultrastructural study of anterograde transport of glial cell line-derived neurotrophic factor from dorsal root ganglion neurons of rats towards the nerve terminal. AB - The glial cell line-derived neurotrophic factor (GDNF) is a potent neurotrophic substance in the central and peripheral nervous systems. The present immunohistochemical study clarified the ultrastructural localization of GDNF immunoreactive substance (GDNF-IR) accumulated at transfected sciatic nerve stumps and also at normal spinal dorsal horn, and has demonstrated that GDNF-IR products appear to be located in dense-cored vesicles within the axons. Furthermore, to determine the source of proximally accumulated GDNF in the transected sciatic nerve, we attempted a transection and a double ligation maneuver involving the sciatic nerve. In the early period after the ligation (20 h), GDNF-IR fibers were observed in the proximal and distal segment of the ligations, but no immunoreactivities were detected in the middle segment. On the other hand, at a late period (8 days) after the transection, GDNF-IR fibers had almost disappeared, but weak GDNF-IR was observed in Schwann cells in the proximal and distal stumps of transected nerve. These findings suggest that most of GDNF-IR was transported from the proximal or distal side in the early period, but was locally synthesized by Schwann cells around the ligations in the late period. Spinal rhizotomy caused prominent accumulation of GDNF-IR products at the cut end of the ganglion side of the dorsal root, but not at the ventral root. These results suggested that dorsal root ganglionic (DRG) sensory neurons are one of the origins of GDNF. The fact that small- to medium-sized DRG neurons show enhanced GDNR-IR after the colchicine treatment may support the above suggestion. In conclusion, the present results strongly suggest that a subgroup of DRG sensory neurons synthesized GDNF-containing dense-cored vesicles in the neuronal somata and anterogradely transports the vesicles to peripheral or central axon terminals. PMID- 11490122 TI - Hepatic indocyanine green uptake and excretion in a rabbit model of steatosis. AB - Transplantation of a fatty liver is associated with a higher incidence of primary non-function of the graft. Indocyanine green (ICG) has been used for assessing hepatic dysfunction but not for quantifying liver steatosis. New Zealand white rabbits were fed a normal diet (group A) or a high-cholesterol (2%) diet for 4, 8, and 12 weeks in groups B, C, and D, respectively. Laparotomy was performed for liver exposure. Hepatic artery, portal vein, and total blood flow, hepatic microcirculation, portal pressure, liver function parameters, and blood cholesterol levels were measured. The hepatic ICG concentration was measured using near-infrared spectroscopy, and its uptake and excretion rates were calculated. The severity of steatosis was assessed from liver biopsy specimens by a semiquantitative grading system. Cholesterol feeding resulted in mild steatosis after 4 weeks and in moderate steatosis after 8 and 12 weeks. Mild steatosis was associated with insignificant changes in haemodynamic parameters, liver function, and ICG handling as compared with controls. Moderate steatosis caused a significant reduction in portal and total hepatic blood flow and microcirculation with a significant increase in hepatic artery flow and portal pressure. These haemodynamic changes were associated with a significant alteration in liver function tests. With moderate steatosis, ICG uptake and excretion were significantly reduced. The ICG uptake rate significantly correlated with total blood flow and microcirculation. The ICG excretion rate significantly correlated with the changes in bilirubin, liver enzymes, and albumin. Direct ICG quantification by near-infrared spectroscopy could be used to assess the severity of hepatic steatosis by reflecting impaired parenchymal perfusion and liver dysfunction. PMID- 11490123 TI - Attenuation of liver and lung injury after hepatic ischemia and reperfusion by a cytokine-suppressive agent, FR167653. AB - BACKGROUND: Hepatic ischemia/reperfusion (I/R) injury is an important clinical problem and leads to the release of the proinflammatory cytokines, TNF-alpha and IL-1. These cytokines play important roles in the induction of polymorphonuclear neutrophil (PMN) activation and infiltration, and induce not only localized hepatic injury but also remote organ injury, especially pulmonary injury. Using a total hepatic ischemia model in rats, we tested our hypothesis that suppression of TNF-alpha and IL-1 by FR167653 ameliorates I/R injury in the liver and lung. METHODS: Male Wistar rats, weighing 240-280 g, were divided into 3 groups, an FR group, a control group and a sham group. In the FR group, FR167653 (1 mg/kg/h) was administered continuously to the animals for 30 min prior to the onset of ischemia and for 2 h after reperfusion. The control group received normal saline. A porto-systemic shunt was placed between the cecal branch of the portal vein and the jugular vein, and total hepatic ischemia was produced for 90 min. The sham group was treated with placement of the porto-systemic shunt only. The 1-week survival rate, liver enzyme activity, hepatic tissue blood flow (HTBF), cytokine mRNA expression, myeloperoxidase (MPO) activity and histological results were studied. RESULTS: The 1-week survival rate and HTBF were significantly higher in the FR group than in the control group. Serum AST, ALT, and LDH levels were significantly lower in the FR group at 30 min, 1 h and 3 h after reperfusion. MPO levels in liver and lung tissue were also significantly lower in the FR group. The expression of IL-1beta mRNA remarkably decreased up to 6 h after reperfusion in the FR group. CONCLUSIONS: We concluded that the inflammatory cytokines, IL 1beta, play important roles in hepatic I/R injury. FR167653 might ameliorate I/R injury and be useful in liver surgery with ischemia. PMID- 11490124 TI - Influence of suture material and technique on end-to-end reconstruction in tracheal surgery: an experimental study in sheep. AB - Resection and end-to-end anastomosis of the trachea represent the preferred treatment for various benign and malignant diseases involving the trachea. Various studies have reported conflicting results with alternative techniques and suture materials for tracheal anastomosis. Our objective was to evaluate three frequently used techniques concerning stenosis rate and histological reaction in a large-animal species. Tracheal resection of 3 cm and end-to-end anastomosis were performed in 15 sheep with the use of three different techniques. In the first group, an interrupted suture with polyglactin, in the second group an interrupted suture with polydioxanone, and in the last group a continuous suture with polypropylene were used. The animals were killed 1, 2, 4, 8, and 24 weeks postoperatively. The luminal stenosis was determined by means of computerized planimetry. All three techniques appeared to be appropriate for tracheal anastomosis. The luminal stenosis developed within the first 8 weeks after surgery. A cross-sectional area of approximately 40-70% was finally achieved. Differences dependent on the suture material are less important than the technical details of the operation. PMID- 11490126 TI - Influence of pentoxifylline and interferon-alpha on prevention of stricture due to corrosive esophagitis. An experimental study in the rat. AB - The effects of pentoxifylline (PTX) and interferon alpha (IFN-alpha) in the prevention of strictures due to corrosive esophagitis in rats were investigated. Forty rats were randomly divided into four equal groups. Corrosive esophagitis was induced in all groups by application of 37.5% NaOH to the distal esophagus for a period of 90 s followed by saline rinse. Histopathologic damage was significantly lower in the PTX and IFN-alpha-treated groups than in the untreated group. During the study period, PTX and INF-alpha-treated animals showed a significant increase in body weight when compared to controls. However, PTX provided more significant prevention of stricture formation than IFN-alpha. In the PTX-treated group, the wall thickness and quantity of hydroxyprolin were significantly lower than in the untreated and IFN-alpha-treated groups. Stenosis index in the PTX group was significantly reduced compared to the control group. PTX prevents the stricture formation due to corrosive esophagitis in this experimental model. IFN-alpha was also shown to prevent stricture formation when considering amelioration of histopathologic damage and increase in body weight. PMID- 11490125 TI - Mechanism of hyporesponsiveness caused by donor-specific transfusion to allogeneic minor antigens. AB - Donor-specific transfusion (DST) is one of the methods to prevent allograft rejection, presumably through induction of donor-specific nonresponsiveness in humans and animal models. In this study, we used a skin graft model in fully allogeneic (AKR into B6), H-2 class I-disparate (bm1 into B6), class II-disparate (bm12 into B6), minor-H-disparate (C3H/SW into B6) and whole MHC-disparate (B10.BR into B10) combinations to analyze the mechanisms of DST-induced immunomodulation from the view point of antigen disparity. Skin graft survival was prolonged by DST only in the combination of bm1 and B6 as already confirmed by many reports. In other combinations, skin graft survivals were not prolonged at all by DST but rather shortened. In a fully allogeneic combination between AKR and B6 in which I-E antigens or Mls-1 antigens are recognized by T-cell subsets with TCR Vbeta11 or Vbeta6, respectively, responses to stimulation by anti Vbeta11 and Vbeta6 monoclonal antibodies were decreased by DST. Furthermore, the number of Vbeta6+ T cells was decreased in the periphery probably due to peripheral clonal deletion. In such a condition, the mixed lymphocyte reactions of B6 to AKR were reduced to some extent, but were clearly detected. In addition, skin allografts from AKR were more rapidly rejected in B6 mice given AKR spleen cells. From these results, DST seems to induce nonresponsiveness to some antigens (I-E antigens, Mls antigens and bm1 antigens in this study), but not to others. Our study also indicated that DST alone, at least in the absence of other treatments, does not contribute to allograft tolerance in fully allogeneic combinations. PMID- 11490127 TI - Synergistic effect of CGS16949A and 5-fluorouracil on a human breast cancer cell line. AB - The effects of the aromatase inhibitor, CGS16949A, and the fluoropyrimidine, 5 fluorouracil (5-FU), on cell cycle distribution and growth were studied using FACS analysis and MTT assay in the human breast cancer cell line, SK-BR-3. CGS16949A induced an increase in the G0-G1 fraction on SK-BR-3 cells, and the growth inhibition rate of the combination of both (65.7 +/- 3.0%) was significantly higher than 10 nM CGS16949A (37.9 +/- 6.9%) or 100 microg/ml 5-FU (45.6 +/- 4.5%); p < 0.01). Administering 5-FU after preincubation with CGS16949A significantly increased the combined cytotoxic efficacy, suggesting that clinical therapy using this combined therapy may be more efficient. PMID- 11490128 TI - Evaluation of splanchnic perfusion and oxygenation during positive end-expiratory pressure ventilation in relation to subcutaneous tissue gases and pH. An experimental study in pigs with oleic acid-induced lung injury. AB - Evaluation of splanchnic perfusion and oxygenation was performed by measurements of serosal tissue oxygen tension (PserO2) and intramucosal pH (pHi) in relation to subcutaneous oxygen tension (PscO2), subcutaneous carbon dioxide tension (PscCO2) and subcutaneous pH (pHsc) in pigs subjected to oleic acid-induced lung injury during ventilation with increasing levels of positive end-expiratory pressure (PEEP). Lung injury resulted in a general hypoxia and redistribution of perfusion away from the subcutaneous and splanchnic tissues, illustrated by a decrease in PaO2 from 93 to 37 mm Hg (p < 0.01), PscO2 from 45 to 17 mm Hg (p < 0.01), PserO2 from 80 to 30 mm Hg (p < 0.01) and pHi from 6.84 to 6.74 (p < 0.05) and a decrease of porta flow from 0.77 to 0.57 l/min. Application of PEEP up to 10-15 cm H2O resulted in an increase of portal vein oxygen tension (PportaO2) from 21 to 34 mm Hg (p < 0.01), PscO2 from 17 to 26 mm Hg (p < 0.05) and PserO2 from 30 to 55 mm Hg (p < 0.05). At PEEP 20 cm H2O PserO2 decreased to 47 mm Hg (p < 0.05). Porta flow decreased continuously with increasing levels of PEEP. PserO2 correlated with PportaO2 (r = 0.7, p < 0.001). pHi correlated poorly with PportaO2 (r = 0.2) and porta flow (r = 0.4). PscO2 and PserO2 correlated well (r = 0.8, p < 0.001). In summary, splanchnic perfusion and oxygenation was better reflected by serosal oxygen tension than pHi in the colon. Changes in serosal oxygenation of the colon paralleled changes in subcutaneous tissue oxygenation. PMID- 11490129 TI - Seroma prevention by using Corynebacterium parvum in a rat mastectomy model. AB - Seroma formation is the most common complication after mastectomy and continues to be an important problem during the early postoperative period. Several surgical and medical methods have been developed to try to overcome this problem; however, so far none have been used successfully in the routine clinical practice. The aim of this study is to evaluate the effects of Corynebacterium parvum (CP) as a sclerosing agent in both prevention and treatment of seromas after mastectomy and axillary dissection in an animal model. Sixty female Sprague Dawley rats underwent mastectomy and axillary dissection under general anaesthesia. Following surgery, the rats were treated in 1 of 3 ways. In the prevention group, 1 cm3 (0.35 mg) CP solution was injected beneath the skin flap just before closure of the incision after mastectomy. In the treatment group, animals in which a seroma was formed, the fluid was aspirated, and 1 cm3 CP solution was injected beneath the flap. In the control group, animals in which seromas formed, aspiration only was performed. The frequency of seroma formation decreased when CP solution was injected immediately after the operation (p < 0.01). In addition, seroma formation was effectively treated by CP injection when compared with the control group (p < 0.05). CP was effective as a prophylactic agent in the prevention group and as a therapeutic agent in the treatment group in this experimental model. CP injection may be useful for the management of this problem in a clinical setting. PMID- 11490130 TI - Behavioural diversity among the wild chimpanzee populations of Bossou and neighbouring areas, Guinea and Cote d'Ivoire, West Africa. A preliminary report. AB - We present a preliminary report on the differences and similarities in material culture among four neighbouring chimpanzee (Pan troglodytes verus) communities. One of these communities includes Bossou, a long-term field site of wild chimpanzees, in Guinea, West Africa. We also conducted surveys of three new sites. Two of those surveyed areas, Seringbara in Guinea and Yeale in Cote d'Ivoire, are located less than 12 km away from Bossou in the Nimba Mountains region, which forms a natural boundary between Guinea and Cote d'Ivoire. The third, Diecke, is situated further south-west, closer to the border with Liberia. During the surveys, we gathered behavioural information about these neighbouring populations of chimpanzees. The differences, as well as similarities, in material culture were tabulated based on our findings. The three behavioural variants found so far involve differences in nut cracking behaviour with regard to the species of nut cracked. Some variation in materials used for nut cracking has also been recorded. However, we still need to establish whether these local variations can be explained by the demands of the physical and biotic environments in which the populations of chimpanzees live. If these alternative hypotheses can be excluded with continuing research at the study sites, these differences are likely to be cultural behaviours that are influenced by the social context and mode, i.e. horizontal, vertical or oblique, of transmission, by the social structure and organisation of each community and/or perhaps by some form of social norms prevalent within these communities. PMID- 11490131 TI - Social organization, reproduction and rearing strategies of Callimico goeldii: new clues from the wild. AB - The callitrichines are a specialized radiation of primates that are characterized in part by variable social systems and cooperative infant care. Callimico goeldii, unlike the other callitrichines, have single rather than twin offspring, reducing the need for allocare and permitting synchronous breeding within groups. Low mortality rates among offspring and unstable social groups are suggested to be possible factors that have led to single births among C. goeldii. Single offspring may benefit from greater maternal investment and more frequent food sharing than twin offspring, factors that may help to explain why C. goeldii reaches sexual maturity more rapidly than other callitrichines. In addition, increased breeding opportunities for young C. goeldii females may have selected for rapid maturation rates among this species. Postpartum ovulation and aseasonal resource availability appear to permit females to have biannual birth seasons, further increasing the potential reproductive output. PMID- 11490132 TI - Using proximity measures to describe mother-infant relationships. AB - What are the relative roles of mothers and infants in maintaining proximity with one another? How do the relative roles of the mother and infant change over time? What do proximity measures tell us about the mother-infant relationship? Questions such as these have been addressed using proximity indices devised by Hinde and coworkers in the 1960s. In this paper, I review how these indices are derived and discuss what they do, and do not, infer about the nature of the mother-infant relationship. An additional index is proposed which indicates whether the mother or the infant is responsible for the majority of changes in proximity. PMID- 11490133 TI - Sequence conservation of nine Barbary macaque (Macaca sylvanus) microsatellite loci: implication of specific primers for genotyping. PMID- 11490134 TI - Cheek pouch use in Papio cynocephalus. PMID- 11490135 TI - Bamboo leaf-based diet of Rhinopithecus bieti at Lijiang, China. PMID- 11490136 TI - Vertical distribution of different age-sex classes in a foraging group of Sichuan golden monkeys (Rhinopithecus roxellana). PMID- 11490137 TI - Lemur responses to diurnal raptor calls in Ranomafana National Park, Madagascar. PMID- 11490138 TI - Foot-rubbing as a multifunctional form of intraspecific communication in Garnett's greater bushbabies (Otolemur garnettii). PMID- 11490139 TI - Group demographics and dispersal in a wild group of Goeldi's monkeys (Callimico goeldii). PMID- 11490140 TI - Pulmonary rehabilitation--the evidence base. PMID- 11490141 TI - Smoking cessation. AB - Smoking cessation has been shown to be cost effective, even in older individuals. It is therefore important that this health promotion is not missed in this vulnerable group. PMID- 11490142 TI - Pulmonary rehabilitation and outcome measures in elderly patients with chronic obstructive pulmonary disease. PMID- 11490143 TI - Salivary concentration of secretory leukocyte protease inhibitor, an antimicrobial protein, is decreased with advanced age. AB - BACKGROUND: Secretory leukocyte protease inhibitor (SLPI) exhibits antimicrobial activities that, in addition to other well-characterized proteins such as lysozyme and lactoferrin, is thought to play a critical role in mucosal defenses. Although elderly individuals are particularly susceptible to mucosal infections, salivary production of SLPI has not been assessed in an aged cohort. OBJECTIVES: Hypothesizing that oral SLPI concentrations are reduced with advanced age, this cross-sectional study compared SLPI concentrations to concentrations of lysozyme, lactoferrin and total protein in unstimulated salivary secretions of healthy, community-dwelling 79+-year-old and younger adults. METHODS: Study participants were 45 non-hospitalized dentate adults aged 79-89 (23 elderly) or 21-51 years (22 non-elderly). Home-based interviews and clinical examinations determined dentate status and confirmed the absence of dentures, oral mucosal disease, anti infective medication use, irradiation therapy for head and neck cancer and self perceived xerostomia. Whole unstimulated saliva was collected from all subjects and analyzed for antimicrobial protein concentration by enzyme-linked immunosorbent assay and for total protein content by the bicinchoninic acid method. Bivariate and multivariate (generalized linear modeling) analyses evaluated the relationships between age, gender and salivary protein concentrations. RESULTS: Mean salivary levels of SLPI and lysozyme were lower in elderly compared with non-elderly subjects (p < 0.001), unlike lactoferrin and total protein levels. Similar results were obtained when concentrations of the individual proteins were normalized to the total protein concentration, suggesting that glandular production of SLPI and lysozyme preferentially decreases with aging. Gender differences were detected only for SLPI concentrations; males had lower SLPI levels than females regardless of age (p < 0.01). Generalized linear models confirmed that age (p < 0.001) and gender (p < 0.05) were each associated with the SLPI concentration and together accounted for 50% of the variation in SLPI concentration in this population. CONCLUSION: These findings indicate that SLPI production is diminished among healthy community dwelling older adults, particularly elderly males. Further investigation should determine the impact of decreased local SLPI production on the increased risk of oral mucosal disease with advanced age. PMID- 11490144 TI - Association of comorbidity with depressive symptoms in community-dwelling older persons. AB - BACKGROUND: Depression in later life poses a serious threat to mental health and well-being of older persons burdened with medical illnesses. Comorbid medical conditions independently, as well as through interactions, may influence the degree of depressive symptoms manifested by the elderly. Insight into the role medical comorbidity plays in the manifestation of depression may help to better address both physical and mental health care needs of the depressed elderly. OBJECTIVE: To examine independent and synergistic effects of comorbid medical conditions in the presentation of depressive symptoms among older adults living in the community. METHODS: Data from a national survey of community-dwelling older persons aged 60 years and over (n = 2,058) in Korea were used. The levels of depression were assessed using an eleven-item Center for Epidemiological Studies Depression Scale. Comorbidity was defined as the number of chronic medical conditions and the combination of disease pairs. Sociodemographic characteristics, self-rated health, physical functioning, history of hospital admission in the past year, frequency of contact with friends or neighbors, and emotional support were used as covariates. RESULTS: Overall, persons with a medical condition compared to those without tended to show a higher depressive symptom score. Comorbidity, in terms of the number of medical conditions, tended to show a significant linear association with depression, controlling for confounders. More important, independent effects of medical conditions as well as a tendency to synergy among specific medical conditions were found, with notable gender differences. CONCLUSION: Given the significant but complex association of comorbid medical conditions with depressive symptoms in aged persons, clinicians should exercise care to address their physical and mental health needs within a common therapeutic context. PMID- 11490145 TI - Physical and sporting activities improve vestibular afferent usage and balance in elderly human subjects. AB - BACKGROUND: Ageing is associated with a reduction in balance, in particular through dysfunction of each level of postural control, which results in an increased risk of falling. Conversely, the practice of physical activities has been shown to modulate postural control in elderly people. OBJECTIVE: This study examined the potential positive effects of two types of regular physical and sporting activities on vestibular information and their relation to posture. METHODS: Gaze and postural stabilisation was evaluated by caloric and rotational vestibular tests on 18 healthy subjects over the age of 60 who regularly practised low-energy or bioenergetic physical activities and on 18 controls of a similar age who only walked on a regular basis. These subjects were also submitted to static and dynamic posturographic tests. RESULTS: The control group displayed less balance control, with a lower vestibular sensitivity and a relatively high dependency on vision compared to the group practising low-energy physical activities, which had better postural control with good vestibular sensitivity and less dependency on vision. The postural control and vestibular sensitivity of subjects practising bioenergetic activities was average, and required higher visual afferent contribution. CONCLUSION: Low-energy exercises, already shown to have the most positive impact on balance control by relying more on proprioception, also appear to develop or maintain a high level of vestibular sensitivity allowing elderly people practising such exercises to reduce the weight of vision. PMID- 11490146 TI - Risk factors of aspiration pneumonia in Alzheimer's disease patients. AB - BACKGROUND: Pneumonia is the reported cause of death of a high proportion of Alzheimer's disease (AD) patients. OBJECTIVES: This study assessed swallowing function to explore the potential risk factors of aspiration pneumonia in AD patients. METHODS: Data were gathered on 121 AD patients (mean age 78.2, SD 7.5, male/female ratio = 48/73) treated in two psychiatric hospitals. Multiple logistic regression analysis was used to estimate odds ratios for the risk of aspiration pneumonia. The following risk factors were considered: age, severity of dementia, silent brain infarction (SBI) in the basel ganglia, use of psychotropic drugs (neuroleptics and benzodiazepines), and gender. The latency time of the swallowing reflex was analyzed (1) among groups based on the severity of dementia; (2) between groups with and without SBI in the basal ganglia; (3) before and after the use of psychotropic drugs in the group without SBI in the basal ganglia, and (4) between males and females. RESULTS: Aspiration pneumonia in ambulatory AD patients was significantly and independently associated with severe dementia, presence of SBI in the basal ganglia, intake of neuroleptics, and the male gender. The mean latency of the swallowing reflex in the severe AD group was significantly longer compared to that in the mild-to-moderate AD group. The mean latency of the swallowing reflex after the intake of neuroleptics was significantly prolonged compared to that obtained before any neuroleptic intake, whereas there was no significant difference in the mean latency of the swallowing reflex before and after benzodiazepine intake. CONCLUSION: We recommend the use of the smallest efficacious dosage of neuroleptics especially in treating severely demented AD patients with basal-ganglia infarction. As soon as the behavioral and psychiatric problems are resolved, drug tapering or discontinuation should be considered in order to prevent life-threatening aspiration pneumonia. PMID- 11490147 TI - Predicting fallers in a community-based sample of people with Parkinson's disease. AB - BACKGROUND: The risk of people with Parkinson's disease (PD) falling is greater than that of the general population but to date, disease-specific predictors of falling have not been identified. OBJECTIVES: To identify one or more features, which would predict individuals at risk of falling during a 3-month prospective follow-up study. METHOD: A battery of standardised tests administered in the home and the laboratory with a 3-month follow-up telephone interview. RESULTS: Sixty three people with PD were recruited from GP practices. Eleven interview variables and six gait laboratory variables were used with subsamples (55 and 44 subjects, respectively) to fit predictive models for identifying future fallers. The number of falls in the previous year was the most important variable, without exception, to be selected as a predictor in various logistic regression models. A history of two or more falls had a sensitivity of 86.4% (95% CI 67.3-96.2%) and a specificity of 85.7% (95% CI 71.2-94.2%) in predicting falling in the next 3 months. CONCLUSION: Healthcare workers should be asking their patients with PD regularly and carefully about falling, and should consider instigating programmes of fall management for patients with PD who have fallen two or more times in the previous 12 months. PMID- 11490148 TI - Natural killer activity and thyroid hormone levels in young and elderly persons. AB - BACKGROUND: On the basis that (1) multiple interactions exist between the hormonal and immune systems, and (2) aging is accompanied by changes in thyroid hormone metabolism and responsiveness, we postulate that thyroid hormones may be involved in the observed decrease in natural killer (NK) activity in a population of apparently healthy elderly subjects. The purpose of the study is to compare NK cytotoxic activity and serum concentrations of TSH and thyroid hormones in healthy old and young people, and to assess in vitro the effects of triiodothyronine (T(3)) on NK activity. MATERIALS AND METHODS: Sixteen of the 47 healthy old people (mean age 64 +/- 5.2) were classified as optimally healthy, and the remainder as 'almost healthy' (according to the criteria of the Senieur protocol) [Ligthart et al., Mech Ageing Dev 1984;28:47-55]; the mean age of the healthy young people was 23.3 +/- 2.3 years. NK cytotoxic activity of peripheral blood mononuclear cells was assessed using (51)Cr release from K562 target cells. The cutoff level for defining low and high NK responses was set at a value of 20%. Serum concentrations of TSH, total thyroxine (T(4)) and total triiodothyronine (T(3)) were measured by radioimmunoassay. RESULTS: NK activity in the 'optimally healthy' elderly was high (mean 41 +/- 12%, SE), whereas 'almost healthy' subjects showed low NK activity (mean 6 +/- 5%). Serum T(4) and TSH levels, but not T(3) concentrations were similar in both the young and old. We observed a significant correlation (r = 0.53, n = 21, p < 0.05) between the serum total T(3) level and the NK activity in the elderly individuals. Under in vitro conditions exogenous T(3) significantly increased NK activity in the elderly subjects who had serum T(3) values at the lower end of the reference range. However, no effect of T(3) on NK activity was observed in peripheral blood mononuclear cells obtained from either old or young individuals who had serum T(3) levels at the midpoint of the range. CONCLUSION: Decreased serum concentrations of total T(3) may contribute to low NK activity in the 'almost healthy' subgroup of the elderly. PMID- 11490149 TI - Two-year decline in vision but not hearing is associated with memory decline in very old adults in a population-based sample. AB - BACKGROUND: Recent cross-sectional research in cognitive aging has demonstrated a robust association between visual acuity, auditory thresholds and cognitive performance in old age. However, the nature of the association is still unclear, particularly with respect to whether sensory and cognitive function are causally related. OBJECTIVE: This study aimed to determine whether marked declines in performance on screening measures of either visual acuity or auditory thresholds have an effect on cognitive decline over 2 years. METHODS: The sample from the Australian Longitudinal Study of Ageing (n = 2,087) were assessed in 1992 and 1994 on measures of sensory and cognitive function as part of a larger clinical assessment. A quasi-experimental design involving comparison of extreme groups using repeated measures MANCOVA with age as a covariate was used. RESULTS: Group performance on measures of hearing, memory, verbal ability and processing speed declined significantly. Decline in visual acuity had a significant effect on memory decline, but not on decline in verbal ability or processing speed. Decline in hearing was not associated with decline in any cognitive domain. CONCLUSION: The common association between visual acuity, auditory thresholds and cognitive function observed in cross-sectional studies appears to be disassociated in longitudinal studies. PMID- 11490150 TI - The spectrum of olive pollen allergens. AB - Olive pollen is one of the most important causes of seasonal respiratory allergy in Mediterranean countries, where this tree is intensely cultivated. Among the high number of protein allergens detected in this pollen, 8 - Ole e 1 to Ole e 8 have been isolated and characterized. Ole e 1 is the most frequent sensitizing agent, affecting more than 70% of the patients suffering of olive pollinosis, although others, such as Ole e 4 and Ole e 7, have also been shown to be major allergens. In this context, the prevalence of many olive pollen allergens seems to be dependent on the geographical area where the sensitized patients live. Some of the olive allergens have been revealed as members of known protein families: profilin (Ole e 2), Ca(2+)-binding proteins (Ole e 3 and Ole e 8), superoxide dismutase (Ole e 5) and lipid transfer protein (Ole e 7). No biological function has been demonstrated for Ole e 1, whereas Ole e 4 and Ole e 6 are new proteins without homology to known sequences from databases. cDNAs encoding for Ole e 1, Ole e 3 and Ole e 8 have been overproduced in heterologous systems. The recombinant products were correctly folded and exhibited the functional activities of the natural allergens. In addition to the Oleaceae family, other species, such as Gramineae or Betulaceae, contain pollen allergens structurally or immunologically related to those of the olive tree. This fact allows to detect and evaluate antigenic cross-reactivities involving olive allergens. The aim of this research is the development of new diagnostic tools for olive pollinosis and new approaches to improve the classical immunotherapy. PMID- 11490151 TI - Microchimerism and systemic sclerosis. AB - Systemic sclerosis (SSc) is a connective tissue disease characterized by progressive fibrosis of the skin and internal organs. SSc is an immunologically mediated disease. A prominent immunological abnormality in SSc patients is the presence of circulating autoantibodies against a variety of nuclear proteins. Furthermore, SSc is characterized by the presence of increased numbers of activated T cells, with the prevalence of CD4+ cells, present in the periphery of skin lesions as well as in other organs in the early stages of the disease. We have recently shown the existence of a predominant activation of IL-4-producing Th2-like T cells in patients with SSc, which may account for the major alterations which occur in this disease. SSc has clinical and serological similarities to chronic graft versus host disease (cGVHD), although there are some important differences. T cells, which orchestrate the tissue damage, are present in great amounts in the inflammatory infiltrates in SSc- and cGVHD affected tissues. More importantly, T cells from cGVHD tissues produce Th2-like cytokines, thus showing a pathogenetic similarity with SSc. SSc has been postulated as a type of cGVHD resulting from the transplacental transfer of cells between mother and fetus. Very recently, we have shown that in SSc, the microchimeric T cells react with the maternal MHC antigens and are able to produce Th2-type cytokines. Both features are characteristics of cGVHD, supporting the hypothesis that SSc is a disease similar to cGVHD. PMID- 11490152 TI - gammadelta T cells as regulators of airway hyperresponsiveness. AB - Airway responsiveness (AR) is determined by complex mechanisms reflecting lung responses to airborne stimuli. Murine studies have identified a number of potential factors modulating AR and thus have contributed to the current understanding of these mechanisms. In allergic inflammation, immune cells, in particular alphabeta T cells, have emerged as important contributors to increased AR. We have found that in contrast to alphabeta T cells, gammadelta T cells can have a negative regulatory effect on AR. Here, we review the current studies on gammadelta T cells in allergic inflammation and discuss their role in modulating AR. We propose that gammadelta T cells exhibit different immune properties depending on the type of stimulus and inflammation. These differential immune properties appear to be associated with specific gammadelta T cell subsets, which control AR to airborne stimuli. In particular, our recent data indicate that the Vgamma4(+) T cell subset acts as an important negative regulator of AR and contributes to maintaining normal lung function in mice. PMID- 11490153 TI - Underdiagnosis and undertreatment of asthma: a 9-year study of Italian conscripts. AB - BACKGROUND: During the last 20 years, the epidemiology of asthma has been investigated in various populations, thus providing interesting data including the accuracy of diagnosis and the adequacy of treatment. METHODS: We studied for 9 years the epidemiological features of asthma in a homogeneous population of 18 year-old males referred to La Spezia Military Navy Hospital for a call-up visit (conscripts). RESULTS: During the period of 1990-1998, 91,700 men were screened at their call-up visit; out of them 5,371 (7%) had asthma, but in 7.4% the disease had not been previously diagnosed. During the observation period there was a significant decrease of the level III severity of asthma, but about one quarter of the asthmatic subjects received no treatment at all. On the other hand, a significant increase in the prescription of inhaled steroids was observed. CONCLUSION: Despite educational efforts, a not negligible underdiagnosis and inadequate treatment of asthma in young adults still persist. PMID- 11490154 TI - Hev b 8, the Hevea brasiliensis latex profilin, is a cross-reactive allergen of latex, plant foods and pollen. AB - BACKGROUND: Plant profilins are important pan-allergens. They are responsible for a significant percentage of pollen-related allergies. Limited information is available about their involvement in the latex-fruit syndrome and the cross reactivities between latex and pollen. We aimed to clone and express the Hevea brasiliensis latex profilin to investigate its allergological significance and serological cross-reactivities to profilins from plant foods and pollens. METHODS: A DNA complementary to messenger RNA (cDNA) coding for the Hevea latex profilin, Hev b 8, was amplified by polymerase chain reaction from latex RNA. Recombinant (r)Hev b 8 was produced in Escherichia coli and used to screen sera from 50 latex- allergic health care workers (HCWs) with well-documented histories of food and pollen allergy and 34 latex-allergic spina bifida (SB) patients. The cross-reactivity of natural Hev b 8 and rHev b 8 with other plant profilins was determined by ELISA inhibition assays. A three-dimensional homology model of Hev b 8 was constructed based on known profilin structures. RESULTS: The cDNA of Hev b 8 encoded a protein of 131 amino acids with a predicted molecular mass of 14 kD. Twelve of the 50 HCWs and 2 of the 34 SB patients were sensitized to Hev b 8. All Hev b 8-sensitized patients showed allergic symptoms to pollen or plant foods. Cross-reactivities between profilins of latex, pollen and plant food were illustrated by their ability to inhibit IgE binding to rHev b 8. Homology modeling of Hev b 8 yielded a structure highly similar to Bet v 2, the birch pollen profilin, with the most distinct differences located at the N-terminus. CONCLUSIONS: We conclude that primary sensitization to latex profilin in the majority of cases takes place via pollen or food profilins. Additionally, pollinosis and food-allergic patients with profilin-specific IgE can be at risk of developing latex allergy. PMID- 11490155 TI - IgG-mediated histamine release from canine mastocytoma-derived cells. AB - BACKGROUND: Recent data suggest that normal tissue mast cells can express functional receptors for IgG under certain conditions. However, little is known about IgG receptor expression and functional consequences in mast cell neoplasms. METHODS: In this study, neoplastic mast cells were obtained from a dog with cutaneous mastocytoma (CM-MC) and from a dog with visceral mastocytoma (VI-MC). Both cell populations were characterized morphologically and functionally. RESULTS: Most cells proliferated constantly in suspension without particular supplements. Doubling times of CM-MC and VI-MC were 52.2 and 27.5 h, respectively. Both cell types were sensitive to formalin fixation, did not contain heparin and were tryptase and chymase positive. Electron microscopy showed fine granules with electron-dense content in both cell populations. The total histamine content of CM-MC and VI-MC was 0.25 and 0.10 pg/cell, respectively. Calcium ionophore A23187 and substance P induced dose-dependent histamine release, whereas compound 48/80 had no effect. Most significantly, both cell types, when sensitized with monomeric dog IgG, released histamine upon stimulation by anti-dog IgG. CONCLUSIONS: Dog mastocytoma-derived cells may be useful to study the regulation of neoplastic mast cell growth and differentiation, as well as IgG receptor-mediated activation in neoplastic mast cells. Further research is required to clarify the pathophysiological significance of constitutive expression of IgG receptors in neoplastic (canine) mast cells. PMID- 11490156 TI - Interleukin-18 is elevated in the sera from patients with atopic dermatitis and from atopic dermatitis model mice, NC/Nga. AB - BACKGROUND: Several lines of in vitro and in vivo studies have demonstrated that interleukin-18 (IL-18) shows both antiallergic and allergy-promoting activities. But its expression in allergic diseases remains unknown. METHODS: Serum IL-18 levels from atopic dermatitis (AD) model mice, NC/Nga and control mice and from patients with AD and healthy volunteers were measured by ELISA. The relationship between IL-18 levels and serum IgE levels or clinical severity was also examined. RESULTS: Serum IL-18 levels from NC/Nga mice were significantly increased compared to those from control mice. The elevation of IL-18 in the sera was observed prior to the onset and during the development of dermatitis in NC/Nga mice. In addition, IL-18 levels in the sera from patients with AD were significantly (p < 0.05) elevated compared to those from healthy volunteers. However, serum IL-18 levels tended to correlate negatively with serum IgE levels in patients with AD and NC/Nga mice. CONCLUSION: IL-18 is overexpressed in AD. PMID- 11490157 TI - Expression and purification of recombinant rat eosinophil-associated ribonucleases, homologues of human eosinophil cationic protein and eosinophil derived neurotoxin, and their characterization. AB - BACKGROUND: Human eosinophils contain two eosinophil ribonucleases, eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). In rats, 8 homologues of human ECP and EDN have been identified. To clarify the biological activity of rat eosinophil ribonucleases, we cloned rat eosinophil-associated ribonuclease (EAR)-1/rat ribonuclease 7 and rat EAR-2/rat ribonuclease 4, and produced recombinant rat pre-EAR-1 and pre-EAR-2 in a bacterial expression system. METHODS: As we have already cloned the complete nucleotide sequence for rat EAR-1, we determined that for rat EAR-2 cDNA by the rapid amplification of cDNA ends procedure. Recombinant rat pre-EAR-1 and pre-EAR-2 were expressed in Escherichia coli as N-terminal 6 x histidine-tagged proteins, isolated from the insoluble fraction of the cell lysate and purified by a single-step method using an Ni-NTA resin column after solubilization with a 6 M guanidine solution. RESULTS: The deduced amino acid sequence revealed that the molecular weight of EAR-2 containing the signal peptide is 17.3 kD and the isoelectric point is 8.59. The homology in amino acid sequence between rat pre-EAR-2, and human pre-ECP and human pre-EDN is 51 and 53%, respectively. The purified and refolded recombinant rat pre-EAR-1 and pre-EAR-2 showed bactericidal activity against E. coli and Staphylococcus aureus. CONCLUSIONS: These findings suggest that rat EAR-1 and EAR 2 act as host defense factors against bacterial infection in rats. PMID- 11490158 TI - Sera from patients with Kawasaki disease induce intercellular adhesion molecule-1 but not Fas in human endothelial cells. AB - BACKGROUND: Kawasaki disease (KD) is an acute vasculitis of unknown etiology occurring in childhood, characterized by abnormalities of the immune system including elevations of proinflammatory cytokines in the serum. We investigated the effect of serum from patients with KD on the expression of intercellular adhesion molecule-1 (ICAM-1) and Fas by human umbilical vein endothelial cells (HUVEC). METHODS: Confluent monolayers of HUVEC were incubated with sera from patients in the acute or convalescent phase of KD. Expression of ICAM-1 and Fas by HUVEC was assessed by flow cytometry. Concentrations of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in sera from patients with KD were measured by an immunoradiometric assay and an enzyme-linked immunosorbent assay, respectively. RESULTS: Sera from patients in the acute phase of KD produced significantly greater ICAM-1 expression by HUVEC than sera from patients in the convalescent phase. In contrast, KD sera did not induce Fas expression. While the mean serum concentration of TNF-alpha in patients in the acute phase of KD was significantly higher than in those in the convalescent phase, IL-1beta concentrations did not differ between the acute and convalescent phases. Exposure of HUVEC to recombinant human TNF-alpha increased the expression of both ICAM-1 and Fas, but a much lower concentration was required for an effect upon ICAM-1. Exogenous TNF-alpha did not induce apoptosis in HUVEC. CONCLUSIONS: These results suggest that increased expression of ICAM-1 by endothelial cells might be involved in the pathogenesis of acute KD, and that TNF-alpha might induce ICAM-1 expression. PMID- 11490159 TI - Antitumor activity of expanded human tumor-infiltrating gammadelta T lymphocytes. AB - BACKGROUND: The objective of this study was to investigate the antitumor activity of selectively expanded gammadelta T cells in tumor-infiltrating lymphocytes (gammadeltaTILs) or tumor ascites lymphocytes (gammadeltaTALs) from patients with colorectal and ovarian epithelial carcinoma (OEC) in vitro and in vivo. METHODS: gammadeltaTILs/TALs were expanded by the solid-phase antibody method; their cytolytic and proliferative activities in vitro were detected by the MTT method and 3H-TdR incorporation and their effect in vivo was evaluated by the nude mice model. RESULTS: Expanded gammadeltaTILs from colorectal tumors demonstrated marked cytotoxicities to allogeneic human colon adenocarcinoma HR8348 and lymphoma Daudi cells, as well as xenogeneic murine thymoma EL-4 cell lines. Cytokines, including IL-2, IL-4, IL-12, IL-15, TNF-alpha and INF-gamma, could promote the cytotoxicities of gammadeltaTILs to tumor cells, whereas IL-10, GM CSF and TFG-beta had no effect on such killing activities. Rested gammadeltaTILs could proliferate strongly in response to mitomycin C-treated Daudi and EL-4 tumor cells, but not to HR8348 tumor cells, suggesting that the latter might possess only cytotoxicity-related antigen recognized by gammadeltaTILs. Either alphabetaTILs or gammadeltaTILs from patients with OEC displayed cytotoxicities to allogeneic or autologous OEC cell lines at a similar strength in vitro. Transferring gammadeltaTILs into Daudi cell-bearing BALB/c nude mice with an injection of IL-2 was able to maintain a high survival rate of the mice for 30 days, when compared with mice treated with alphabetaTILs or without any treatment (p < 0.05). Without coinjection of IL-2, after 3 months of Daudi tumor inoculation, a high survival rate was observed in gammadeltaTIL-treated mice. Similarly, adoptive gammadeltaTALs from the ascites of patients with OEC transferred into nude mice displayed a stronger antitumor response to OEC SKOV3 cells than alphabetaTALs in vivo. Tumor volumes in gammadeltaTAL-treated mice were smaller than in alphabetaTAL-treated or non-TAL-treated mice within the period from day 23 to day 50 after tumor inoculation (p < 0.05). Fifty days after SKOV3 tumor inoculation, a decreasing trend of carcinogenic rate was observed in gammadeltaTAL-treated nude mice. CONCLUSION: Taken together, our results suggest that gammadeltaT cells could be a new candidate for adoptive immunotherapy in the future treatment of patients with cancer. PMID- 11490160 TI - Food-dependent exercise-induced anaphylaxis: clinical and laboratory findings in 54 subjects. AB - BACKGROUND: In some subjects, specific foods trigger anaphylaxis when exercise follows ingestion (specific food-dependent exercise-induced anaphylaxis, FDEIAn). Skin test and/or RAST positivity to foods suggest an IgE-mediated pathogenic mechanism. Others suffer from anaphylaxis after all meals followed by exercise, regardless of the food eaten (nonspecific FDEIAn). We sought to identify the culprit foods with a diagnostic protocol. METHODS: We collected detailed histories and performed skin prick tests (SPT) with 26 commercial food allergens, prick plus prick tests (P+P) with 15 fresh foods (including 9 assessed with SPT), and RAST for 31 food allergens. Treadmill stress tests were administered after a meal without any positive food (food plus exercise challenge, FEC). RESULTS: Among the 54 patients, 6 could not recall any suspect food. The other 48 suspected a specific food in at least one episode. The most frequent were tomatoes, cereals and peanuts. Fifty-two subjects were positive to at least one food (22 to more than 20), whereas 2 showed no positive results. All suspect foods were positive. SPT, P+P and RAST displayed different degrees of sensitivity. Each test disclosed some positivities not discovered by others. Two subjects reacted to FEC. Overall, 48 patients probably had specific FDEIAn and the other 6 nonspecific FDEIAn. CONCLUSION: It is useful to test both in vivo and in vitro an extensive panel of foods. Avoidance of foods associated with skin test and/or RAST positivity for at least 4 h before exercise has prevented further episodes in all our patients with specific FDEIAn. PMID- 11490161 TI - Multiple sclerosis and ultraviolet radiation: time to shed more light. PMID- 11490162 TI - Regional variation in multiple sclerosis prevalence in Australia and its association with ambient ultraviolet radiation. AB - The aim of this study was to conduct an ecological analysis of the extent to which ultraviolet radiation (UVR) levels might explain the regional variation of multiple sclerosis (MS) in Australia. MS prevalence data for six Australian regions were compared with UVR levels of the largest city in each region, with some other climatic variables and with the melanoma incidence in the same regions. A close association was found between the theoretical MS prevalence predicted from UVR levels and the actual prevalence. Furthermore, the negative correlation between UVR and MS prevalence (r = -0.91, p = 0.01) was higher than the positive correlation observed for UVR and malignant melanoma incidence (r = 0.75, p = 0.15 for males and r = 0.80, p = 0.10 for females). This study demonstrated that the regional variation in MS prevalence in the continent of Australia could be closely predicted by regional UVR levels. It is consistent with the hypothesis that UVR exposure may reduce the risk of MS possibly via T lymphocyte-mediated immunosuppression. Analytical epidemiology studies are required to investigate this specific hypothesis. PMID- 11490163 TI - Multiple sclerosis and ionizing radiation. AB - The etiology of multiple sclerosis (MS) may involve exposure to infectious, chemical or physical agents damaging the blood-brain barrier and an autoimmune reaction against myelin breakdown products. Here we report a pooled analysis of 174 MS cases and 815 population controls from two case-control studies with regard to such a potentially damaging exposure, namely X-ray examinations, radiological work and treatment with ionizing radiation. Exposure was assessed by questionnaires to the subjects. We obtained odds ratios of 4.4 (95% confidence interval, CI, 1.6-11.6) and 1.8 (95% CI 1.2-2.6) for radiological work and X-ray examinations, respectively; 5 cases, but no controls, in one of the studies had been treated with ionizing radiation. Our data and some other observations reported in the literature suggest a contributory role for ionizing radiation to the development of MS in some cases. PMID- 11490164 TI - The incidence of vascular dementia in Canada: a comparison with Europe and East Asia. AB - Vascular dementia (VaD) is the second commonest subtype of dementia after Alzheimer's disease (AD). However, its incidence has been studied much less extensively than that of AD. This article reviews the incidence data for VaD reported in the international literature. Results from 10 incidence studies are compared to those of the Canadian Study of Health and Aging using age standardized incidence ratios (SIR). SIRs vary from 0.42 to 2.68, indicating that geographical variation is still present after taking into account the countries' differential age distributions. It is still unclear if these differences are due to genetic and/or environmental factors since a large part reflects methodological differences between studies. PMID- 11490165 TI - Prognosis of Alzheimer's disease: the Rotterdam Study. AB - The aim of this study was to construct a prognostic model to predict the progression of Alzheimer's disease (AD). Prevalent and incident cases with AD came from the Rotterdam Study, a population-based prospective cohort study of persons aged 55 years and older, including those living in institutions. Rate of cognitive decline, as measured by the Mini Mental State Examination (MMSE score), was predicted by a random effects model. Risk of institutionalization and death were estimated with polytomous logistic regression analysis. At baseline, 306 subjects were diagnosed with prevalent AD and had complete data on living conditions and cognitive function. After a mean follow-up of 2.1 years, 95 subjects with incident AD had been diagnosed. Prevalent patients showed a slower decline in cognitive function than incident patients (p = 0.004). For prevalent and incident AD patients, high age and low cognitive performance were the strongest predictors for institutionalization and death. These prognostic risk functions can provide information on the decline of Alzheimer patients and might be used to better evaluate the effect of treatments for AD. PMID- 11490166 TI - Delay between stroke onset and emergency department evaluation. AB - BACKGROUND: Public educational programs have been developed to reduce delays between the onset of ischemic stroke symptoms and emergency department evaluation. An increase in the proportion of patients presenting soon after stroke would reflect the effectiveness of these efforts. METHODS: All patients (n = 506) with ischemic stroke admitted to an academic medical center located within the 'Stroke Belt' of the USA were prospectively identified over 2 years (1998 1999). Demographics, stroke characteristics and time from symptom onset to arrival in the emergency department were recorded. RESULTS: A higher proportion of ischemic stroke patients presented within 3 h of symptoms in 1998 than in 1999 (18% of 234 vs. 8% of 272, p = 0.0001). Those with less severe strokes (Canadian Neurological Scale score; Spearman r = 0.18, p < 0.0001) and younger patients (r = -0.09, p = 0.04) had greater delays. There was no difference in time to presentation based on race (13% of whites and blacks presented within 3 h, p = 0.70) or sex (16% of women vs. 9% of men, p = 0.10). Logistic regression showed that time to presentation was independently related to both stroke severity and year. CONCLUSIONS: These data show that, after accounting for other variables, the proportion of stroke patients presenting within 3 h of symptom onset to one academic medical center decreased by 10% between 1998 and 1999. Revision of public stroke-related educational programs may need to be considered. PMID- 11490168 TI - Estimation of mortality and morbidity due to strokes in India. AB - In order to control the stroke problem, its magnitude should be assessed. India is ranked among the countries where the information on stroke is minimal. We decided to review the information available in order to estimate the mortality and morbidity due to stroke in India. Information was collected through electronic search, hand search and contact with experts. Each article was reviewed for relevance and epidemiological rigor. The demographic data were as derived from published government figures. The prevalence from individual studies was pooled and weighted based on sample size. Analysis was done separately for males and females at 10-year intervals (20 years onwards). A total of 7 studies was located, but 2 were discarded. All were done in rural areas except 2 which also included urban areas. The prevalence was estimated as 203 per 100,000 population above 20 years amounting to a total of about 1 million cases. The male to female ratio was 1.7. Around 12% of all strokes occurred in population below 40 years. The estimation of stroke mortality was seriously limited by the method of classification of cause of death in the country. The best estimate derived was 102,000 deaths; which represented 1.2% of total deaths in the country. There is need to initiate steps to collect data on morbidity and mortality due to stroke in the country as a first step towards control measures. PMID- 11490167 TI - Stroke in the urban population of Calcutta--an epidemiological study. AB - A population-based cluster survey on stroke disorders was conducted for the first time in the city of Calcutta, India. The population surveyed totaled 50,291. The crude prevalence rate of stroke was 147/100,000 (age-adjusted prevalence 334/100,000). The annual incidence rate of stroke for the year 1998-1999 was 36/100,000 (age-adjusted annual incidence rate 105/100,000). Women outnumbered men regarding stroke prevalence in all age groups except in the 50- to 69-year age group. There were relatively more cases of cerebral haemorrhage in our study, compared to those in the western countries. Case-control analysis found hypertension to be the most significant risk factor for stroke. PMID- 11490169 TI - Prevalence of Parkinson's disease in Bulgaria. AB - Despite some publications on Parkinson's disease prevalence in Bulgaria, its frequency in the country still remains uncertain. Earlier studies in Bulgaria were carried out on large population groups and were based on medical records without further diagnostic confirmation. The aim of the present study was to perform an epidemiological investigation on Parkinson's disease in Bulgaria, using strict diagnostic criteria in an attempt to obtain a more accurate estimate of its frequency in this country. We studied the prevalence of Parkinson's disease in two small regions of Bulgaria. Cases were ascertained by the medical records and confirmed by personal examination. The prevalence of Parkinson's disease was found to be 164.2 per 100,000 with 47 cases in the first region and 169.8 per 100,000 with 155 cases in the second region. Based on two small population studies, the prevalence ratio in different parts of Bulgaria is not significantly different and is similar to other European countries. PMID- 11490171 TI - Selective handling of information in patients suffering from restrictive anorexia in an emotional Stroop test and a word recognition test. AB - Several studies have been carried out using the Stroop test in eating disorders. Some of these studies have brought to light the existence of cognitive and attention deficits linked principally to weight and to food in anorexic and bulimic patients. The aim of the current study is to replicate and to clarify the existence of cognitive and attention deficits in anorexic patients using the Stroop test and a word recognition test. The recognition test is made up of 160 words; 80 words from the previous Stroop experiment mixed at random and matched from a semantic point of view to 80 distractions. The recognition word test is carried out 2 or 3 days after the Stroop test. Thirty-two subjects took part in the study: 16 female patients hospitalised for anorexia nervosa and 16 normal females as controls. Our results do not enable us to confirm the existence of specific cognitive deficits in anorexic patients. PMID- 11490172 TI - Study of family history in seasonal affective disorder. AB - OBJECTIVE: In our investigation we assessed the risk of morbidity for psychiatric disorders among the first-degree relatives of patients with seasonal affective disorders (SAD) and compared it with a control group of patients suffering from nonseasonal mood disorders (NSMD). METHODS: Over a period of 12 months (June 1994 to May 1995) we recruited patients consecutively admitted to our psychiatric university outpatient clinic in a prospective study. All patients were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, revised 4th edition. A total of 344 patients presented themselves with a diagnosis of affective disorder. Out of these, 36 were diagnosed as having SAD. From the same group of 344 patients, we selected a matched control group of 36 patients suffering from NSMD. The experimental and control groups were matched according to sex, age, severity of illness and number of siblings. RESULTS: There was no significant difference concerning the lifetime prevalences for psychiatric disorders among the fist-degree relatives in both groups (SAD = 16.5% and NSMD = 19%). CONCLUSION: It seems that there is no difference in familiarity for psychiatric disorders between SAD and NSMD. PMID- 11490173 TI - Caudate nucleus dopamine D(2) receptors in depressed suicide victims. AB - Several lines of evidence indicate the involvement of the dopamine system in depressive states. In this post-mortem study, the binding of [(3)H]raclopride to dopamine D(2) receptors in the caudate nucleus was investigated in 13 depressed suicide victims and 19 controls. There were no differences in B(max) or K(d) between the two groups. A subgroup consisting of individuals with major depression, however, had significantly higher K(d) values than controls. Previous findings regarding changes in dopamine metabolism in depression and antidepressant effects of dopamine agonists seem, according to the present study, not to be reflected by alterations in density or affinity of dopamine D(2) receptors in depressed suicide victims. PMID- 11490174 TI - HPA axis dysfunction in major depression: relationship to 5-HT(1A) receptor activity. AB - Major depression is associated with a dysfunction of the serotonergic activity and the hypothalamic-pituitary-adrenal (HPA) axis. Moreover, a reciprocal relationship between the serotonergic and HPA axis systems has been hypothesized. The purpose of the present study was to assess the relationship between sensitivity of 5-HT(1A) receptors as measured by hormonal (ACTH, cortisol and PRL) and temperature responses to flesinoxan and HPA axis activity as measured by the dexamethasone suppression test. The sample included 21 inpatients with major depression. Dexamethasone nonsuppressors exhibited lower ACTH responses to flesinoxan as compared with dexamethasone suppressors. The results showed that a dysfunction in 5-HT(1A) receptor activity could be due to a hypersecretion of cortisol. PMID- 11490175 TI - Pharmacological characterization of the serotonin transporter in young and elderly subjects. AB - The potency of some tricyclics (imipramine and clomipramine) and selective 5-HT reuptake inhibitors (fluoxetine, paroxetine, and citalopram) in displacing the [(3)H]paroxetine binding to platelet membranes was measured in young and elderly subjects of both sexes. The results showed that the most potent compound in all subjects was paroxetine, followed by clomipramine, citalopram, fluoxetine, and imipramine, with no differences between male and female subjects. All drugs, except paroxetine and clomipramine, showed significantly lower pKi values in the elderly subjects of both sexes. These findings would suggest that although the pharmacological profile of the 5-HT transporter is not modified qualitatively by age, quantitative changes in its affinity do perhaps occur which would justify more careful studies on this topic in order to get optimal dosages of drugs acting at this level. PMID- 11490176 TI - Intraspinal pressure influences CSF disposition of tryptophan and 5-HIAA. AB - Cerebrospinal fluid (CSF) concentrations of monoamine compounds are influenced by factors such as age, gender, height, body weight, tapping time, and atmospheric pressure. We have now examined the role of intraspinal pressure. Thirteen male volunteers underwent lumbar puncture in the right decubitus position without preceding strict bed rest. The intraspinal pressure was recorded, and monoamine precursors, transmitters, and metabolites were analyzed in two consecutively collected CSF fractions. Tryptophan in 12 ml of CSF and the 5-hydroxyindoleacetic acid concentration ratio [fraction II (7--12 ml CSF)/fraction I (0--6 ml CSF)] correlated with the intraspinal pressure. Hypothetically, the intraspinal pressure may be a confounding factor for a correct interpretation of CSF tryptophan and 5-hydroxyindoleacetic acid concentrations, and this is an issue that has to be addressed in future CSF studies. PMID- 11490177 TI - Neurotrophic factor S100 beta in major depression. AB - Disturbances in the serotonergic system are considered to be implicated in the pathophysiology of depressive disorders. The possible role of the neurotrophic factor S100 beta, which is suspected to regulate regeneration of serotonergic synapses, has not been investigated in depressive disorders. The S100 beta concentration in the cerebrospinal fluid was measured in 11 patients with the current diagnosis of mild or moderate depressive episodes (DSM-IV) and in 11 matched control patients. Using the t test for paired samples, the presence of a depressive episode was significantly associated with an elevation of the cerebrospinal fluid concentration of S100 beta (t = 2.6, d.f. = 10, p = 0.024). Replications of this finding in severely depressed patients are necessary to confirm the association between neurotrophic factor S100 beta and depressive disorders. PMID- 11490178 TI - Alpha-2-adrenoreceptors in depressed suicide attempters: relationship with medical lethality of the attempt. AB - Several lines of evidence tend to suggest a role for noradrenaline, and more specifically alpha-2-adrenoreceptors, in the biology of suicidal behavior. The purpose of this study was to assess the growth hormone (GH) response to clonidine, an alpha-2-adrenergic agonist, in majorly depressed inpatients with a history of highly lethal suicide attempt compared to depressed patients with a history of low lethal suicide attempt and nonattempters. Our sample included 20 male depressed inpatients with a history of suicide attempt compared to 20 male depressed nonattempters. We did not observe any significant difference between suicide attempters and nonattempters for GH peak values (2.4 +/- 2.9 vs. 4.1 +/- 3.7 ng/ml; F = 2.52, d.f. = 1, 38, p = 0.12). Moreover, GH peak responses to clonidine were not related to the degree of lethality of the attempt. The results of the present study do not support a major role for noradrenaline in the biology of suicidal behavior. PMID- 11490179 TI - Association between the Ser9Gly polymorphism of the dopamine D3 receptor gene and tardive dyskinesia in Chinese schizophrenic patients. AB - It has been suggested that dopamine D3 receptor (DRD3) may have important implications for antipsychotic-induced tardive dyskinesia (TD). Previous studies have demonstrated an association between a serine to glycine polymorphism in the first exon of the DRD3 gene and TD; however, the results have been inconsistent. Therefore, we have replicated these studies using a Chinese sample population. A total of 115 schizophrenic patients from chronic wards were assessed for TD severity using the Abnormal Involuntary Movements Scale (AIMS) and were subsequently genotyped for the DRD3 polymorphism. The mean AIMS score for patients carrying the heterozygote (DRD3(ser-gly)) was significantly greater than for those with the homozygotes (DRD3(ser-ser) and DRD3(gly-gly)). Our results are in line with a previous report, the results of which suggest that the presence of the DRD3(ser-gly) genotype may be a risk factor for the development of TD in patients treated with antipsychotics. PMID- 11490180 TI - Chronic treatment with clozapine, but not haloperidol, increases striatal ecto-5' nucleotidase activity in rats. AB - In the search for differential mechanisms underlying clozapine's superior antipsychotic efficacy, the purinergic system has been considered, since an antagonist of the adenosine receptor A(2A) was shown to block clozapine acute effects on c-fos expression in rat striatum. Further investigating the interaction of clozapine with the purinergic system, we studied the effects of chronic treatment (28 days, intraperitoneal) with clozapine (25 mg/kg) and haloperidol (1.5 mg/kg) on the activity of ectonucleotidases in the striatum and hippocampus of rats. Clozapine selectively increased striatal 5'-nucleotidase activity (22%) compared to control and haloperidol groups. In vitro, neither drug affected enzyme activities. These results reinforce the differential effects of clozapine compared to haloperidol on the purinergic system. PMID- 11490181 TI - Inhibition of K(+)-evoked release of rat striatal 5-hydroxytryptamine by an atypical antidepressant: trazodone. AB - Using microdialysis, extracellular concentrations of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were determined in the striatum of rats. In rats given trazodone, m-chlorophenylpiperazine dihydrochloride, or imipramine, the concentrations of 5-HT were unchanged. 5-HIAA in trazodone- or imipramine treated rats, however, was respectively, decreased to 80 or 65% of preinjections levels. When the potassium concentration (K(+)) was increased up to 150 mmol/l in the perfusate, the concentrations of 5-HT increased to about ten times the basal levels in the rats given saline. In rats treated with trazodone, K(+)-evoked elevations of 5-HT were less than five times the basal level. Multiple trazodone administrations prolonged the duration of inhibition of 5-HT release. In rats treated with other drugs, the K(+)-evoked 5-HT release was not affected. These observations suggest that trazodone itself might reduce 5-HT neural transmission. PMID- 11490182 TI - Spatial structure of brain electric fields during intermittent photic stimulation. AB - EEG changes in 27 young healthy male right-handed volunteers on intermittent photic stimulation (IPS) were estimated using global field power (GFP), EEG microstate modeling and analysis (EMMA), and low-resolution electromagnetic brain tomography (LORETA). The GFP significantly increased at flashing frequency and high harmonics. Three model maps were extracted with the EMMA procedure, from which high alternation rates of each microstate were observed. Moreover, two of the three model maps contributed very highly, occurring most frequently. LORETA imaging of the three model maps obtained from the EMMA procedure showed that both visual dominant cortical areas were activated, especially in the left hemisphere. These results suggest that IPS does not cause peculiar spatial configurations of the brain electric field, but does cause acceleration and deviation of the microstate alternation. Also, a functional laterality between hemispheres might be enhanced by symmetric IPS. PMID- 11490183 TI - Vertex epidural hematoma with communicating bifrontal subgaleal hematomas treated by percutaneous needle aspiration. AB - The case of an 11-year-old boy is presented who suffered a bicycle accident with a parasagittal skull fracture, a small vertex epidural hematoma, frontal contusions and a frontal subgaleal hematoma. Enlargement of the vertex epidural hematoma was diagnosed after development of a slight paraparesis on day 11 with the aid of MRI. Three percutaneous needle aspirations of the subgaleal hematoma with a total of 59 ml being evacuated led to quick recovery and disappearance of the subgaleal as well as the vertex epidural hematoma. It is speculated that both hematomas communicated via the skull fracture thus making the evacuation of the epidural hematoma by subgaleal punctures possible. PMID- 11490184 TI - Abnormal coagulation during pediatric craniofacial surgery. AB - INTRODUCTION: This prospective study of children undergoing major craniofacial surgery was undertaken to determine whether abnormal hemostasis occurred and to characterize any coagulopathy found. METHODS: Coagulation tests, blood loss and blood product transfusions were recorded perioperatively. Packed red blood cells (PRBC) were transfused to maintain target hematocrit. Patients with blood loss >100 ml/kg (group A, n = 5) were compared to patients with blood loss <100 ml/kg (group B, n = 22) using Mann-Whitney U test (p < 0.05). RESULTS: Twenty-seven children (age range 2.9--27.9 months) had median total blood loss of 64 ml/kg. At completion of surgery, median coagulation values differed significantly between groups for prothrombin time (A: 16.6 s; B: 13.8 s), partial thromboplastin time (A: 44 s; B: 29 s), thrombin time (A: 28 s; B: 23 s), thromboelastograph reaction time (A: 7 mm; B: 4 mm), prothrombin fragment F1.2 (A: 1.9 nmol/l; B: 3.3 nmol/l) and platelet count (A: 174 K/mm(-3); B: 239 K/mm(-3)). Fibrinolysis was not associated with blood loss. Median units transfused were in group A 3 units and group B 1 unit (p = 0.001). All patients received PRBC transfusions but only group A patients received other blood products (fresh frozen plasma, platelets). CONCLUSION: Children transfused with PRBC during craniosynostosis repair can become coagulopathic from coagulation factor depletion when hemorrhage approaches 1.5 times estimated blood volume. PMID- 11490185 TI - The Influence of hair shave on the infection rate in neurosurgery. A prospective study. AB - OBJECTIVE: To investigate whether not shaving hair in neurosurgical operations carries an increased infection rate. METHODS: Taking advantage of different practices among neurosurgeons in a single institution, we embarked upon a prospective non-randomised study of 100 consecutive neurosurgical procedures involving 90 paediatric patients aged 7 days to 16.8 years. The patients were split into two groups ('hair shave' and 'no hair shave'). The differences with respect to wound complications, positive microbiology on wound culture swabs and wound infection rates were analysed. Other factors considered were the cleansing solution, prophylactic antibiotic regime, duration of the operation, the surgeon's experience and the patient's age. RESULTS: The only complications observed were 4 incidences of wound dehiscence (2 in the hair shave and 2 in the no hair shave group) and 3 shunt infections (2 in the hair shave and 1 in the no hair shave group). We did not find any significant difference between the two arms for any of the factors assessed. Age was a significant factor in shunt infection, as all shunt infections were seen in patients aged less than 6 months, regardless of whether the hair was shaved or not (p = 0.024, Fisher's exact test). CONCLUSION: This study confirms our clinical experience that no hair shave is a good alternative to the traditional hair shaving approach, allowing patients to enjoy the psychological benefits of undisturbed body image while recovering from major surgery. PMID- 11490186 TI - Aspergillus spinal epidural abscess. AB - Spinal abscess due to Aspergillus is rare. A young boy with chronic granulomatous disease and aspergillosis of the rib had been treated with antifungal treatment 3 months earlier. The patient presented with a brief history of progressive paraparesis. Imaging showed D9--11 vertebral involvement and destruction of the D10 vertebral body with angulation and a large dorsally placed, multiloculated epidural abscess extending from D6 to L2. There was also extensive granulation anterior to and on either side of the vertebrae. The patient underwent extensive laminectomy and decompression of all the loculi and partial removal of the granulation tissue. Aggressive medical treatment was started. The authors recommend an aggressive surgical and medical approach in such cases of disseminated invasive aspergillosis, even though the result may not be very satisfactory. This report discusses the full clinical profile and management of Aspergillus spinal epidural abscess and emphasizes the need to follow up these cases to detect recurrence and new lesions, even if the patients are on adequate medical treatment. In spite of all efforts, high morbidity and mortality is common in such patients. PMID- 11490187 TI - Sudden death from fulminant acute cerebellitis. AB - Acute cerebellitis is postulated to result from viral and/or autoimmune etiologies. This disease has been reported to have a variable course. We report a case of sudden death from acute fulminant cerebellitis in a 13-year-old ballet dancer. Serial CT and MRI demonstrated rapid progression of the disease. Histopathologic correlation is provided. The etiologies, clinical course, therapeutic interventions and postmortem evaluation of this potentially life threatening condition are briefly reviewed. PMID- 11490188 TI - Spinal intramedullary ependymal cyst. Report of two cases and review of the literature. AB - Two pediatric patients (4 and 5 years of age) with spinal intramedullary ependymal cysts located at the cervical and dorsal cord are reported here. One patient was admitted with subtle signs, while the other had disabling spastic quadriparesis. In both patients, MRI depicted a well-demarcated, localized, nonenhancing intramedullary lesion isointense with CSF on T(1)- and T(2)-weighted images. Total excision of the covering membrane of the cyst was not possible in either case. The child with spastic quadriparesis had a remarkable recovery, while the subtle signs of the other patient regained normalcy at the follow-up of 4 and 5 months, respectively. PMID- 11490189 TI - Neurosurgical interventions in children with Maroteaux-Lamy syndrome. Case report and review of the literature. AB - This paper reports the case of a 14-year-old child with Maroteaux-Lamy syndrome (mucopolysaccharidosis type 6) who was treated consecutively for compressive damage of the optic nerves, hydrocephalus communicans and progressive spastic tetraparesis within 2 years. The clinical course of the patient is presented and the pathophysiologic mechanisms of disease progression in patients with Maroteaux Lamy syndrome are discussed and reviewed. PMID- 11490190 TI - Maternal lung adenocarcinoma metastatic to the scalp of a fetus. Case report. AB - Maternal malignancy metastatic to the fetus is a rare event, with most neoplasms being either melanocytic or hematopoietic in origin. This report is the first known case of a maternal lung adenocarcinoma metastatic to a fetus. At 2 months of age, this male infant developed multiple scalp masses that were locally resected but rapidly recurred. The histology of the scalp lesions and that of a biopsy of the mother's tumor were both adenocarcinoma and were remarkably similar in appearance. In situ hybridization of tumor cells from this male infant found many large nuclei with XX signals indicating that the tumor was of maternal origin. This patient is alive, well and free of malignancy 5 years after a wide local resection of the scalp and skin grafting. PMID- 11490191 TI - Membranous obstruction of the fourth ventricle outlet. A case report. AB - We report the case of a 15-year-old girl with amenorrhea and a several-week history of headache. After the diagnosis of membranous obstruction of the foramen of Magendie suggested by MRI, suboccipital craniotomy for removal of the membrane was carried out. The patient made an excellent postoperative recovery, and postoperative phase-contrast MRI demonstrated patent cerebrospinal fluid (CSF) pathways at the level of the foramina of Magendie and Luschka. We believe that this case is of interest because of the unequivocal evidence on MRI studies of the occlusion of the foramen of Magendie preoperatively, and because of the dramatic postoperative MRI findings demonstrating the effectiveness of the surgical procedure both in terms of ventricular size and CSF flow characterization. PMID- 11490192 TI - Colloid cyst and epidermoid cyst. PMID- 11490193 TI - Orbital growing fracture. PMID- 11490194 TI - Vein of galen aneurysmal malformation. PMID- 11490195 TI - Urinary incontinence and ventriculoperitoneal shunts. PMID- 11490196 TI - A seven-week-old infant with fever, CSF pleocytosis and intraspinal mass. PMID- 11490197 TI - Inhibitory effect of a TP-receptor antagonist, S-1452, on antigen-induced nasal plasma exudation in guinea pig model for allergic rhinitis. AB - S-1452, a selective thromboxane (Tx) A(2) receptor (TP-receptor) antagonist, was evaluated in antigen- and U-46619 (a TxA(2) mimetic)-induced guinea pig nasal plasma exudation models. Exposure of the nasal cavity of actively sensitized guinea pigs to aerosolized ovalbumin (OA) caused marked exudation of dye into both the nasal mucosa and nasal airway lumen. These responses were significantly inhibited by S-1452 (30 mg/kg, p.o.) as well as an H(1)-antihistamine, diphenhydramine (5 mg/kg, i.v.). In addition, exposure of the nasal cavity of nonsensitized guinea pigs to aerosolized U-46619 or histamine also resulted in nasal plasma exudation, and S-1452 (1 mg/kg, p.o.) almost completely suppressed the U-46619-induced response but did not affect the histamine-induced one, even at a high dose of 30 mg/kg. These results indicate that TxA(2) as well as histamine may play an important role in antigen-induced nasal plasma exudation in guinea pigs, and S-1452 can be expected to be useful for the treatment of allergic rhinitis. PMID- 11490198 TI - Effects of bergenin, the major constituent of Mallotus japonicus against D galactosamine-induced hepatotoxicity in rats. AB - The hepatoprotective effects of bergenin, a major constituent of Mallotus japonicus, were evaluated against D-galactosamine (GalN)-induced liver damage in rats. Bergenin (50, 100 and 200 mg/kg) was given orally once daily for 7 successive days and then GalN 400 mg/kg was injected intraperitoneally to rats at 24 and 96 h after the final administration of bergenin. Pretreatment with bergenin reduced the increased enzyme activities of alanine/aspartate aminotransferase, sorbitol dehydrogenase, gamma-glutamyltransferase and the elevated level of malondialdehyde induced by GalN. Bergenin restored the decreased hepatic contents of glutathione as well as the decreased activities of glutathione S-transferase and glutathione reductase by GalN towards normalization, suggesting that the hepatoprotective effects of bergenin may consist in maintaining adequate levels of hepatic glutathione for the removal of xenobiotics. The present results indicate that bergenin has hepatoprotective effects against GalN-induced hepatotoxicity in rats. PMID- 11490199 TI - The Na(+)/H(+) exchanger SM-20220 attenuates ischemic injury in in vitro and in vivo models. AB - The aim of this study is to clarify whether the activation of a Na(+)/H(+) exchanger (NHE) is tightly concerned with neuronal and glial cell injury induced by ischemia using a selective NHE inhibitor, SM-20220 (N-(aminoiminomethyl)-1 methyl-1H-indole-2-carboxamide methanesulfonate). Two hours of hypoxia followed by 24 h of reoxygenation induced lactate dehydrogenase (LDH) release, a marker of cell membrane damage, in cultured neurons and glia derived from rats. SM-20220 significantly reduced LDH release in both cells in a concentration-dependent manner, and this effect was statistically significant at concentrations of more than 10(-8) mol/l for neurons and 10(-7) mol/l for glia. A standard NHE inhibitor, 5-(N-ethyl-N-isopropyl)-amiloride, also reduced LDH release in neurons at concentrations of more than 10(-7) mol/l. In a rat transient middle cerebral artery occlusion model, intravenous infusion of SM-20220 reduced cerebral infarction when the serum concentration of SM- 20220 was maintained at about 10( 7) mol/l. These results suggest that the activation of the NHE plays an important role in ischemic neuronal and glial cell injury, and NHE inhibitor may have good therapeutic value for the treatment of ischemic stroke. PMID- 11490200 TI - Characterization of Ascaris-induced biphasic skin allergic reaction model in mice: possible roles of mast cells in early-phase and CD4-positive T cells in late-phase reactions. AB - We established an Ascaris-induced biphasic skin allergic reaction in mice. In the early-phase reaction (EPR), mast cell degranulation was observed, and tranilast inhibited ear edema. In mast-cell-deficient mice (WBB6F(1)-W/W(V) mice), ear edema in the EPR disappeared, whereas that in the late-phase reaction (LPR) remained. Eosinophils increased, and CD4-positive T cells tended to increase in the LPR. Anti-CD4 antibody, anti-IL-4 antibody and anti-IL-5 antibody all inhibited ear edema and had a tendency to inhibit eosinophil infiltration in the LPR. These data suggest that the EPR is induced by histamine released from mast cells, whereas the LPR is induced by IL-4 and IL-5 produced from CD4-positive T cells. PMID- 11490201 TI - Alteration of drug kinetics in rats following exposure to trichloroethylene. AB - The effect of trichloroethylene (TCY) was investigated to determine whether repeated exposure alters the pharmacokinetics of some drugs. Sprague-Dawley rats were given intraperitoneal injections of TCY (5 mmol/kg) in corn oil once daily for 3 days, while the control group received only corn oil. Four hours after the last dose, theophylline, quinidine, or pentobarbital were administered. Blood samples were collected at appropriate intervals for drug analyses. There was a small decrease in plasma clearance of theophylline, with no change in volume of distribution (V(d)) as compared with controls. For quinidine, the elimination half-life was unchanged, and the V(d) was decreased by 40%. The clearance of pentobarbital was decreased by 40% in male rats, but not in the females. Nonetheless, the duration of the sleeping time for both sexes was remarkably prolonged as compared with the control group. There was a decrease in the cytochrome P-450 content only in male rats. In conclusion, exposure to TCY causes changes in some drug kinetics, probably resulting from differential effects on the drug-metabolizing enzymes. PMID- 11490202 TI - Gastroprokinetic effect and mechanism of SK-896, a new motilin analogue, during the interdigestive period in conscious dogs. AB - SK-896 [(Leu(13))motilin-Hse] is a new human motilin analogue synthesized from Escherichia coli using a biotechnological method. We investigated the gastrointestinal motor-stimulating effect of SK-896 and the mechanism of this effect using implanted force transducers in conscious dogs. Infusion of SK-896 during phase I in the interdigestive state induced interdigestive migrating contractions like motility in the gastroduodenum. The motility index (MI(0-20)) of gastric antrum motor activity induced by SK-896 was increased dose dependently (r = 0.830, p < 0.001), and the MI(0-20) induced by SK-896 at a dose of 0.25 microg/kg/h for 20 min was the same as that for spontaneous phase III contractions. The SK-896-induced MI(0-20) was significantly decreased by atropine, hexamethonium, dopamine, granisetron, and yohimbine. Conversely, ketanserin, phentolamine, timolol, and naloxone did not have significant effects on SK-896-induced MI(0-20). The effects of these drugs on human motilin (0.25 microg/ kg/h for 20 min) induced MI(0-20) were the same as those of SK-896. These results indicate that SK-896 induces gastrointestinal motility during the interdigestive period in dogs with regulation of acetylcholine release from the cholinergic nerve terminal via the parasympathetic nervous system in the same fashion as human motilin. PMID- 11490203 TI - Angiotensin-induced enhancement of excitatory junction potentials evoked by periarteriolar nerve stimulation and vasoconstriction in rat mesenteric arteries are both mediated by the angiotensin AT1 receptor. AB - The aims of the present study were to determine the angiotensin II (AngII) receptor subtype(s) involved in vasoconstriction and enhancement of sympathetic neurotransmission in rat isolated mesenteric arteries. Vasoconstriction was assessed in mesenteric artery ring preparations suspended under 0.5 g of tension in a myograph. In control arteries, with an intact endothelium, AngII (1 nmol/l-3 micromol/l) caused a concentration-dependent contraction. The pEC(50) for AngII was 7.6 +/- 0.2 and the maximum response of 0.24 +/- 0.07 g was reached with 100 nmol/l. In the presence of indomethacin (3.0 micromol/l) and N(omega)-nitro-L arginine (NOLA) (100 micromol/l) to remove the influence of endothelium-derived prostaglandins and nitric oxide, the maximum response evoked by AngII was increased to 0.48 +/- 0.1 g and the pEC(50) was 7.6 +/- 0.3. The AT1 receptor antagonist losartan (30 nmol/l) competitively blocked the AngII-induced contractions with an estimated pA(2) of 8.2 in both the control arteries and in arteries treated with indomethacin and NOLA. The AT2 receptor antagonist PD 123319 (1 micromol/l) did not affect AngII-induced contractions under either condition. Conventional intracellular microelectrode recording techniques were used to investigate the effects of AngII on excitatory junction potentials (EJP) evoked by stimulation of periarteriolar sympathetic nerves. Stimulation with trains of 10 pulses delivered at 0.9 Hz evoked EJP which were blocked by tetrodotoxin (0.1 micromol/l), guanethidine (30 micromol/l) and the P(2X) receptor desensitizing agent alpha,beta-methylene ATP (30 micromol/l) suggesting the EJP were mediated by ATP, or a related purine, released from sympathetic nerves. AngII (0.3- 100 nmol/l) did not affect the resting membrane potential or the amplitude of the first EJP, but did enhance the amplitude of the plateau EJP later in the train. A maximum 49.2 +/- 3.9% enhancement of the plateau EJP amplitude was elicited by 10 nmol/l AngII and the pEC(50) was 9.1 +/- 0.1. The facilitatory effect of AngII on EJP amplitude was not altered in the presence of indomethacin and NOLA. Losartan (30 nmol/l) competitively blocked the AngII induced enhancement of plateau EJP amplitude, with an estimated pA(2) of 8.6. PD 123319 did not alter the enhancement of plateau EJP amplitude by AngII. The results from the present study show that both the vasoconstriction and enhancement of plateau EJP amplitude by AngII in rat mesenteric arteries are blocked by the AT1 receptor antagonist losartan and are unaffected by the AT2 receptor antagonist PD 123319. PMID- 11490204 TI - In vitro and in vivo activity of new rhodium (III) complexes against Leishmania donovani. AB - The activities of 17 new rhodium drug complexes were determined against Leishmania donovani promastigotes. The five most active salts were selected: [Rh(III)(2-amino-6-ethoxybenzothiazole)(4)Br(2)](+)Br(-); [Rh(III)(2 bromothiazole)(4)(Br)(2)](+)Br(-); [Rh(III)(mefloquine)(4)(Cl)(2)](+)Cl(-); [Rh(III)(2-mepacrine)(4)(Cl)(2)](+)Cl(-), and [Rh(III)(oxamniquine)(4)(Cl)(2)](+)Cl(-), which induced growth-inhibition rates of more than 50% at 24 h of treatment and at the maximum dosage tested. The cytotoxicity assays on the macrophage cell line J-774 showed high cytotoxicity for the salts [Rh(III) (mefloquine)(4)(Cl)(2)](+)Cl(-), [Rh(III)(2 mepacrine)(4)(Cl)(2)](+)Cl(-) and [Rh(III)(oxaminquine)(4)(Cl)(2)](+)Cl(-) with a percentage of specific (15)Cr release of 49.3, 64.8 and 53.2% at 24 h of incubation and 100 microg/ml. Meanwhile, assays of the other compounds showed practically no cytotoxicity. The ultrastructural studies in the flagellates treated with the salt [Rh(III)(2-amino-6-ethoxybenzothiazole)(4)Br(2)](+)Br(-) showed some alterations in the nucleus of the parasites with a very condensed chromatin and an electrodense endosome. This compound showed a high in vivo activity in parasitized Wistar rats. PMID- 11490205 TI - Distribution of P2X(1) and P2X(3) receptors in the rat and human urinary bladder. AB - Adenosine 5'-triphosphate (ATP) is known to play a significant role as a neurotransmitter in smooth muscle. There is evidence to show that ATP can cause bladder contractions and may also be involved in the processing of sensory information in the urinary bladder. These effects are likely to be mediated by P2X receptors, namely P2X(1) and P2X(3), respectively. This study set out to investigate their distribution in rat and human urinary bladders. P2X(1) receptor immunoreactivity was found on detrusor muscle fibres and P2X(3) receptor immunoreactivity was found in the urothelium of both species. This is the first demonstration of a non-neuronal localisation for P2X(3) receptors. No clear evidence was found for the presence of P2X(3) receptors on calcitonin gene related peptide-containing sensory nerves and therefore P2X(3) receptors may not have a direct role in the mediation of sensory responses to ATP in the urinary bladder. PMID- 11490206 TI - Prognostic value of renal cell carcinoma nuclear grading: multivariate analysis of 333 cases. AB - OBJECTIVE: To evaluate the independent predictive value of the nuclear grading system according to Fuhrman in relation to the disease-specific survival of patients with renal clear cell carcinoma. MATERIAL AND METHODS: 333 patients who underwent radical nephrectomy for renal clear cell carcinoma between 1983 and 1999 were evaluated. In all patients we retrospectively studied nuclear grading, average tumor size, multifocality, pathologic stage of primary tumor, vein invasion, lymph node involvement and distant metastases. The Kaplan-Meier method was applied to evaluate disease-specific survival rates. The log rank test was used to compare survival curves and for univariate analysis. The Cox proportional hazards model was used for the multivariate analysis. RESULTS: Histologic grade was G1 in 83 cases (25%), G2 in 117 cases (35%), G3 in 110 cases (33%) and G4 in 23 cases (7%). Our data showed that nuclear grading according to Fuhrman is related to medium tumor size (p < 0.0001), pathologic stage of cancer (p < 0.001), venous system invasion (p < 0.001), lymph node involvement (p < 0.001) and distant metastases (p < 0.001). The disease-specific survival after 5 and 10 years was 94 and 88%, respectively, in patients with G1, 86 and 75% in patients with G2, 59 and 40% in patients with G3 and 31% in patients with G4 (log rank p value < 0.0001). Multivariate analysis showed that nuclear grading by Fuhrman has a prognostic independent predictive value (hazard ratio = 1.8461, p = 0.002). CONCLUSIONS: Nuclear grading is an important independent predictive factor of disease-specific survival in patients with renal cell carcinoma. PMID- 11490207 TI - Risk factors for the development of bladder transitional cell carcinoma following surgery for transitional cell carcinoma of the upper urinary tract. AB - To determine the risk factors for development of transitional cell carcinoma (TCC) of the bladder (BTCC) following surgery for TCC of the upper urinary tract (UUT-TCC) in patients without history of BTCC, 85 patients surgically treated for UUT-TCC (34 female, 51 male; median age 66, range 42-85 years) were reviewed retrospectively. The Cox proportional hazards model was used to assess the association of relevant clinicopathologic factors with BTCC-free survival in patients without a history of BTCC and TCC-specific survival in all. Median follow-up duration was 35 (range 1-193) months. Six patients (7%) had previous histories of BTCC, and 6 others (7%) had concurrent BTCC at the time of surgery for UUT-TCC. Of 70 patients who had no history of BTCC and underwent follow-up cystoscopy, 24 (34%) developed BTCC during follow-up after surgery. Univariate analysis identified female sex, postoperative systemic chemotherapy, and incomplete distal ureterectomy as significant risk factors for new development of BTCC. After multivariate analysis adjusted for age and pathological (p) T stage in the TNM classification, all three factors remained significant, with respective hazard ratios of 5.56 (95% confidence interval (CI), 1.99-15.6; p = 0.001), 3.19 (95% CI, 1.34-7.62; p = 0.009) and 2.99 (95% CI, 1.08-8.26; p = 0.03). Only pT stage was a significant independent risk factor for TCC-specific death. Female sex and postoperative systemic chemotherapy, as well as incomplete distal ureterectomy, are possible riks factors for development of BTCC following surgery for UUT-TCC. PMID- 11490208 TI - Natural history of refluxing distal ureteral stumps following upper tract surgery in children with ectopic ureter or ureterocele. AB - BACKGROUND: It is a difficult problem to manage refluxing distal ureteral stumps after an upper tract approach in the ectopic ureter and ureterocele. We evaluated the natural history of the remaining refluxing distal ureteral stumps through a close follow-up. PATIENTS AND METHODS: From 1991 to 1998, nine patients with the ectopic ureter or ureterocele, whose age ranged from one to 20 (median 5.0) months, had undergone an upper tract surgery for ectopic ureter (2) or ectopic ureterocele (7). The upper tract surgery for ureterocele or ectopic ureter included a total nephrectomy (2), a partial nephrectomy (6) or a proximal ureteroureterostomy (1), leaving distal ureteral stump. We evaluated the clinical results of the remaining ureteral stumps every 6 or 12 months. The postoperative follow-up period was 51 (median; range 21-96) months. RESULTS: Among the 9 patients, 5 showed disappearance of the reflux and a markedly reduced stump size in the follow-up study. Improvement was noted in 4 patients within 6 months and in 1 by 20 months postoperatively. Two patients subsequently underwent a bladder level surgery due to problems not associated with the ureteral stump itself. Two patients have been followed up for 30 and 39 months, but without any improvement. An intermittent asymptomatic bacteriuria was noted in all patients during the follow-up, but no patient suffered from symptomatic urinary tract infection requiring an additional surgery. CONCLUSIONS: Since refluxing distal ureteral stumps were spontaneously resolved in more than half of the cases and did not cause significant problems during the follow-up, it is not necessary to remove the refluxing ureter simultaneously at the time of the upper tract surgery. PMID- 11490209 TI - Endocrine alteration in the adrenal gland in kidney transplant patients. AB - INTRODUCTION: Endocrine alterations associated with chronic renal failure have been reviewed recently. Some of these alterations are of clinical relevance. The aim of this study was to investigate the effect of renal transplantation on the endocrine system of the adrenal gland of the transplant recipients. METHODS: The serum angiotensin-converting enzyme (SACE), plasma renin (PR) and plasma aldosterone (PA) were examined in 30 patients before and after renal allotransplantation. Additionally measured parameters were blood pressure, serum creatinine, potassium, sodium, the duration of dialysis and immunosuppressive medication. RESULTS: Six weeks after renal transplantation, serum creatinine decreased from 820.07 +/- 172.01 to 138.12 +/- 67.54 micromol/l. In the same period, serum potassium decreased from 5.42 +/- 0.89 to 4.17 +/- 0.42 mmol/l. PA and PR decreased from 1,150.84 +/- 976.06 to 233.52 +/- 217.07 micromol/l, and from 121.07 +/- 100.12 to 26.16 +/- 10.86 microU/ml, respectively. SACE decreased from 0.21 +/- 0.21 to 0.13 +/- 0.11 micromol/l. No significant correlation was seen with blood pressure, serum sodium, the duration of dialysis and immunosuppressive drugs. Additionally, 2 patients with acute renal graft dysfunction showed significant increases in PR and PA. After successful treatment both levels declined very quickly to prerejection levels. Patients after binephrectomy show no elevation in PR (5-47 microU/ml) or PA (21-416 micromol/l) neither before nor after renal transplantation. CONCLUSIONS: We conclude that renal transplantation has profound effects on the recipient's renin-angiotensin aldosterone system. Because of the rapid depression after renal transplantation, it does not appear to be involved in the pathogenesis of post transplantation hypertension but may reflect a role for repair processes after renal allotransplantation. PMID- 11490210 TI - DNA methylation analysis in immature testicular sperm cells at different developmental stages. AB - Testicular sperm extraction and intracytoplasmic sperm injection (TESE-ICSI) is a frequently used therapeutic option in azoospermic males. Genetic imprinting is a mechanism of gene regulation - mediated by the methylation of deoxyribonucleic acids (DNA) - by which only one of the parental copies of a gene is expressed. Whether the establishment of the genetic paternal imprint in this immature haploid sperm cell is complete and stable at this stage of maturation has never been analysed. In the present study, a highly sensitive heminested methylation specific polymerase chain reaction for the target region 15q11-13 was developed for imprinting analysis at the single-cell level. Imprinting analysis was then carried out on DNA extracted from single diploid leucocytes (n = 25), ejaculated spermatozoa (n = 88), elongated testicular spermatids (n = 30), and round spermatids (n = 25). Amplification was obtained in 57% of the ejaculated spermatozoa, in all elongated spermatids and in 20% of the round spermatids. In the amplified samples, only the paternal imprint was detected. The maternal imprint was not found at all in any of the sperm cells; in 56% of the diploid leucocytes, the maternal and paternal imprints were detected at the same time (complete failure in the other 44%). The data reveal the completed establishment of the correct paternal imprint in ejaculated spermatozoa, elongated spermatids and amplified round spermatids. However, the high rate of amplification failure in round spermatids remains a factor of uncertainty. PMID- 11490211 TI - Effect of terazosin on the lipid profile in patients with symptomatic benign prostatic hyperplasia. AB - INTRODUCTION: To determine the changes in plasma lipid levels in symptomatic benign prostatic hyperplasia (BPH) patients receiving terazosin treatment. MATERIALS AND METHODS: The study included 99 patients with BPH aged 44-74 years. The patients were divided into 3 groups: in group 1 (n = 25) with baseline total cholesterol levels of >220 mg/dl, terazosin 5 mg/day was used; in group 2 (n = 56) with basal total cholesterol levels of < 220 mg/dl, terazosin 5 mg/day was used, and group 3 (n = 18) did not use terazosin and was defined as the control group. Plasma levels of total cholesterol, low-density lipoprotein, high-density lipoprotein and triglyceride were recorded, and the high-density lipoprotein to total cholesterol ratio was calculated at the beginning of the study and after 12 weeks. RESULTS: The total cholesterol level decreased from the baseline level by 10.88% after 12 weeks (p < 0.05) in group 1. The decrease was observed in 22 of 25 patients (88%). In group 1, the mean plasma total cholesterol level decreased significantly (p < 0.05), but the decrease was not significant in group 2 and no change was observed in group 3. The mean plasma low-density lipoprotein level decreased significantly in group 1 (p < 0.05), but no change was observed in the other 2 groups. The mean plasma high-density lipoprotein level increased in group 1, whereas no change was observed in the other 2 groups. The mean plasma triglyceride level decreased significantly in groups 1 and 2 (p < 0.05), but no change was observed in group 3. The high-density lipoprotein to total cholesterol ratio increased significantly in group 1, but no change was observed in the other 2 groups. CONCLUSION: We suggest that terazosin may be a reasonable choice because of the beneficial effect on the lipid profile in older symptomatic BPH patients with a higher ratio of dyslipidemia. PMID- 11490212 TI - Objective penile vascular response to intraurethral prostaglandin E2 (dinoprostone). AB - PURPOSE: Intraurethral prostaglandin E2 (PGE2) administration is a noninvasive treatment modality for erectile dysfunction. The purpose of this study was to evaluate the objective effects of this agent by measuring peak systolic velocities of cavernosal arteries after intraurethral PGE2 administration and comparing with the results obtained with an intraurethral placebo gel and intracavernous papaverine injection. MATERIALS AND METHODS: The study group consisted of 22 consecutive impotent volunteers with a mean age of 46 years who had normal penile arterial responses as determined by penile arterial responses on papaverine-stimulated penile duplex ultrasonography. The peak systolic velocity in cavernosal arteries was recorded after intracavernous injection of 60 mg of papaverine. All patients received 1 mg of intraurethral PGE2 gel and placebo at 15-day intervals. The peak systolic velocities were recorded after each treatment. RESULTS: Mean peak systolic velocity achieved by intraurethral administration of PGE2 gel (25 +/- 8 cm/s) was less than that achieved by intracavernous papaverine (40 +/- 6 cm/s) but higher than that obtained by placebo (15 +/- 4 cm/s). Twelve patients had erections, while 9 had partial and 1 had no erection with intraurethral PGE2. Placebo did not cause any erections. No serious side effects were observed. CONCLUSION: Intraurethral administration of PGE2 appears to be an effective and simple method for increasing penile arterial flow and can be used during penile Doppler ultrasonography to stimulate the penile arterial system. PMID- 11490213 TI - Spontaneous erections in a patient with erectile dysfunction after palliative chemotherapy for non-small cell lung cancer. AB - Erectile dysfunction is a common problem, especially among older men. It is often caused by psychological problems, and is also the reason for pronounced impairment of psychosocial well-being. Many systemic diseases, genitourinary surgery, drugs, particularly antihypertensive and psychotropic drugs, and also chemotherapeutic agents and dexamethasone are attributed as being causes of erectile dysfunction. In our case, severe erectile dysfunction was present for 8 months before non-small cell lung cancer was diagnosed. Normal sexual function, observed for a short period immediately following chemotherapy, is a highly unusual finding and has not been published before. Chemotherapeutic agents have repeatedly been shown to result in cessation of sexual function including erection. While we cannot offer a definite explanation for our findings, undefined paraneoplastic processes leading to erectile dysfunction amenable to successful cytotoxic intervention could be a possible explanation for our observation. PMID- 11490214 TI - Solitary cervical lymph node manifestation of a seminoma without detectable primary. AB - The rare case of a 65-year-old male with a cervical lymph node manifestation as the only tumour manifestation is reported. The diagnostic and therapeutic management of the case is discussed. This represents the fourth case with such a condition reported in the literature. PMID- 11490215 TI - Renal angiomyolipoma extending into the right atrium. AB - Renal angiomyolipomas (AMLs) are benign renal tumors that may possess the features of a malignant neoplasm, such as local and vascular invasion. We describe the diagnosis and management of a rare case of AML associated with tumor thrombus extending into the right atrium. PMID- 11490216 TI - Percutaneous resection of renal transitional carcinoma: venous injury and its conservative management. AB - An interesting observation encountered at percutaneous resection of a renal transitional cell carcinoma and its conservative management is described. During resection of the tumour sudden heavy haematuria was encountered. Nephroscopic inspection revealed the blood to be coming from behind a loop-generated flap, the raising of which allowed entry of the endoscope into a sizeable vein and thence upwards into the renal vein and inferior vena cava (IVC). On table nephrostogram confirmed contrast outlining the renal vein and IVC. Placement and clamping of a nephrostomy tube was followed by stabilization of the patient and the bleeding stopped. The urine became clear within 24 h. Later on repeat nephrostogram showed an intact pelvicalyceal system with an acceptable tumour clearance. This case highlights the importance of a trial of conservative management in the treatment of bleeding complications during nephroscopy. PMID- 11490217 TI - Skin involvement from micropapillary bladder carcinoma as the first clinical manifestation of metastatic disease. AB - Cutaneous metastases from transitional cell carcinoma (TCC) of the bladder occur infrequently. We report the case of a 68-year-old man presenting with an isolated skin nodule a few weeks after local excision of a micropapillary bladder carcinoma. This rare variant of TCC, which is strikingly reminiscent of the histological pattern of ovarian papillary serous carcinoma, is associated with poor prognosis and apparently high metastatic potential. PMID- 11490218 TI - Rapidly relapsing squamous cell carcinoma of the renal pelvis associated with paraneoplastic syndromes of leukocytosis, thrombocytosis and hypercalcemia. AB - A case history is reported here in which leukocytosis, thrombocytosis and hypercalcemia associated with rapidly relapsing squamous cell carcinoma (SCC) of the renal pelvis were observed. In a 58-year-old man, SCC of the renal pelvis was documented during nephrolithotomy, and right nephrectomy was performed. Local relapse of the tumor occurred rapidly in 2 months' time and hypercalcemia, leukocytosis and thrombocytosis worsened in accordance with tumor volume. Cranial computerized tomography (CT), thorax CT and bone scintigraphy were negative for metastasis. The serum parathyroid hormone level was 28 pg/ml (normal 9- 55 pg/ml). To disclose leukocytosis and thrombocytosis, peripheral smear and bone marrow aspiration were performed and no pathologic finding regarding any hematologic disorder was found; the samples were also BCR-ABL negative and Philadelphia chromosome negative. Production of several factors by tumor cells may be responsible for this paraneoplastic syndrome. The association of SCC of the renal pelvis with this triple paraneoplastic syndrome is an extremely rare occurrence. PMID- 11490219 TI - Multidisciplinary treatment of advanced testicular tumor with bulky liver metastasis. AB - A 21-year-old man with far-advanced nonseminomatous germ cell tumor of the left testis is presented. He had multiple bulky metastases in the liver and retroperitoneum with an extraordinarily elevated serum alpha-fetoprotein (23,500 ng/ml). He received multidisciplinary treatment consisting of systemic chemotherapy, cytoreductive left hepatic lobectomy, percutaneous ablation therapy, transarterial chemoembolization, and external beam irradiation for median segments of the liver. The efficient combination treatment normalized the tumor markers within 6 months and has maintained complete serological remission for 4.7 years. PMID- 11490220 TI - Alpha-fetoprotein-producing renal cell carcinoma. AB - Alpha-fetoprotein (AFP) is recognized as a tumor marker of yolk sac tumors, liver cancer and some other cancers of the digestive organs. Renal cell carcinoma (RCC) producing AFP is a rare entity. A case of AFP-producing RCC with solitary bone metastasis, but without liver involvement, is reported. The stain specific to AFP proved the presence of AFP in the cytoplasms of more cells of the renal tumors. Additionally, the other published cases are reviewed. These cases indicate that mesoderm-originating malignant tumors such as RCCs can produce AFP in some situations. So, AFP is probably more universal than believed, although it is generally a popular and useful tumor marker for hepatocellular carcinomas and yolk sac tumors. PMID- 11490221 TI - Excessive verumontanum hyperplasia causing infertility. AB - Retrograde ejaculation is an uncommon cause of infertility in otherwise healthy men. We hereby report a previously undescribed verumontanum hyperplasia as a cause of retrograde ejaculation in a 32-year-old male that was successfully treated by transurethral resection. PMID- 11490222 TI - Prostatic abscess due to Candida with no systemic manifestations. AB - Prostatic abscess due to fungi is a rare condition. It is generally secondary to systemic disease in immunosuppressed patients. It usually occurs with affection of other organs in a septic patient. Only in exceptional cases does it occur isolatedly. We present the case of a prostatic abscess due to Candida albicans with no systemic manifestations. The diagnosis is helped by transrectal ultrasound, which allows to differentiate this condition from nonabscessed acute prostatitis. The treatment of choice is ultrasound-guided transrectal needle aspiration after antibiotic therapy has been started. As with abscesses of bacterial origin, an ultrasonographic follow-up is required due to the possibility of persistence or recurrence. PMID- 11490223 TI - Genitourinary complications of sickle cell disease. AB - PURPOSE: In the last half century the molecular biology, pathophysiology and natural history of sickle cell disease have been well defined. Sickle cell disease causes microvascular occlusion, which is manifested in most organ systems. The genitourinary tract is most commonly affected by hematuria, urinary tract infection and priapism but other more serious sequelae have been identified. MATERIALS AND METHODS: We performed a computerized MEDLINE search from 1965 to the present and a bibliographic review of cross references. These references were analyzed for meaningful findings and case reports. RESULTS: The diagnosis and management of sickle cell disease have advanced rapidly with a significant increase in the life expectancy of affected patients and recognition of a greater number of genitourinary complications. Renal function may be mildly altered or lost completely. Patients with sickle cell disease are at increased risk for urinary tract infection. Priapism is a painful complication of sickle cell disease that is poorly understood and challenging to treat and prevent. Testicular infarction has also been noted. Furthermore, renal medullary carcinoma, a highly lethal tumor, develops almost exclusively in young patients with sickle cell trait. CONCLUSIONS: Heightened awareness of the genitourinary complications of sickle cell disease may prevent end stage disease, including renal failure and impotence. New forms of therapy for sickle cell disease, such as hydroxyurea, may prevent these complications in the future. PMID- 11490224 TI - Lymphatic mapping and detection of sentinel nodes in patients with bladder cancer. AB - PURPOSE: We examined the possibility for detecting sentinel nodes in patients with bladder cancer and whether the histopathological status of identified sentinel nodes reflected that of the lymphatic field. MATERIALS AND METHODS: A total of 13 patients with bladder cancer who met the criteria qualifying them for radical cystectomy had intravesical injections of radioactive tracer and blue dye marker around the tumor followed by lymphoscintigraphy to visualize lymphatic drainage and detect sentinel nodes. Sentinel nodes were identified preoperatively by the blue color and increased radioactivity and were compared histopathologically with other routinely excised lymph nodes. RESULTS: Sentinel nodes were detected in 85% (11 of 13) of patients. There were 4 patients who had sentinel nodes containing tumor cells, and each metastasis was only seen in the detected sentinel node. There were no false-negative sentinel nodes. Of the metastatic sentinel nodes 3 were located outside the normally excised lymph nodes of the obturator fossa. CONCLUSIONS: Sentinel nodes can be detected in patients with bladder cancer. The histopathological status of the identified sentinel nodes was diagnostic for all other excised lymph nodes. Sentinel nodes often seem to be located outside the obturator lymphatic field, which is normally examined during preoperative staging of bladder cancer. PMID- 11490225 TI - Incisional hernia and its repair with polypropylene mesh in renal transplant recipients. AB - PURPOSE: We evaluate the incidence of incisional hernia after kidney transplantation, predisposing factors and the results of surgical repair with polypropylene mesh. MATERIALS AND METHODS: We reviewed the records of 371 consecutive kidney transplants performed between April 1995 and February 2000. Patients with clinical signs of hernia at the transplant incision site were included in the study. Predisposing factors for incisional hernia were also reviewed. A prospective protocol of surgical correction was established using polypropylene mesh and patient outcome was studied. RESULTS: We identified 14 patients (3.8%) with an incisional hernia at the transplant incision site. Hernias developed 3 to 840 days after transplant surgery and were significantly more common in white (p = 0.019) and cadaveric graft (p = 0.02) recipients. Predisposing factors in 11 cases included complications of transplant surgery in 7, bladder obstruction in 2, large polycystic kidneys in 1 and chronic pulmonary disease in 1. Surgical repair was performed by primary fascial approximation and polypropylene mesh reinforcement in 13 cases and by pre-peritoneal mesh placement in 1. Minor subcutaneous wound infection developed in 1 patient. No relapses were noted at a mean followup of 17.8 months. CONCLUSIONS: In the majority of cases incisional hernia develops in the first 3 months after transplant surgery. The incidence is significantly higher in white patients and after cadaveric donor transplantation. Surgical complications of transplant surgery are important predisposing factors for incisional hernia after kidney transplantation. Surgical repair using polypropylene mesh is safe and effective in this group of patients. PMID- 11490226 TI - Eight-year experience with transperitoneal laparoscopic adrenal surgery. AB - PURPOSE: Laparoscopic adrenalectomy is currently the technique of choice for removing benign adrenal lesions. Various laparoscopic techniques and approaches have been reported using the transperitoneal or retroperitoneal approach. We present our 8-year experience with and long-term results of transperitoneal laparoscopic adrenalectomy. MATERIALS AND METHODS: Between October 1992 and October 2000, 161 laparoscopic approaches to the adrenal gland were performed, including 145 unilateral and 10 bilateral adrenalectomies, and 6 conservative operations. Patients were placed in the 60-degree flank position with the bed flexed to increase the surgical field. To avoid hypertensive crisis, especially in patients with pheochromocytoma, the first step involved early ligation of the adrenal vein. RESULTS: The laparoscopic procedure was successfully completed in all except 4 cases, which were converted to open surgery. Mean operative time was 160 minutes in the unilateral, 245 in the bilateral and 90 in the conservative group. Delayed complications included hemoperitoneum in 3 patients, which was drained surgically, severe blood loss in 3 treated with blood transfusion and wound infection in 2. Patients were ambulatory on the morning of postoperative day 1 and were discharged home 2.8, 5 and 1.8 days after unilateral, bilateral and conservative surgery, respectively. CONCLUSIONS: Laparoscopic transperitoneal adrenalectomy is a safe, effective, minimally invasive approach in patients with benign functioning or nonfunctioning adrenal masses. This technique involves low morbidity, minimal postoperative analgesic requirements and a short hospital stay. PMID- 11490227 TI - Clinical role of F-18 fluorodeoxyglucose positron emission tomography for detection and management of renal cell carcinoma. AB - PURPOSE: We evaluate the accuracy of F-18 fluorodeoxyglucose (FDG)-positron emission tomography (PET) for staging and management of renal cell carcinoma. MATERIALS AND METHODS: FDG-PET was performed in 25 patients with known or suspected primary renal tumors and/or metastatic disease and compared with conventional imaging techniques, including computerized tomography (CT). Histopathological confirmation was obtained in 18 patients and confirmation of the disease was by followup in the remainder. The impact of FDG-PET on disease management was also assessed. RESULTS: Of the 17 patients with known or suspected primary tumors FDG-PET was true positive in 15, true negative in 1 and false negative in 1. Comparative CT was true positive in 16 patients and false-positive in 1. The accuracy of FDG-PET and CT was similar (94%). All patients would have undergone radical nephrectomy after conventional imaging findings but FDG-PET results altered treatment decisions for 6 (35%), of whom 3 underwent partial nephrectomy and 3 avoided surgery due to confirmation of benign pathology or detection of unsuspected metastatic disease. Of the 8 cases referred for evaluation of local recurrence and/or metastatic disease FDG-PET changed treatment decisions in 4 (50%), with disease up staged in 3 and recurrence excluded in 1. Compared with CT, FDG-PET was able to detect local recurrence and distant metastases more accurately and differentiated recurrence from radiation necrosis. CONCLUSIONS: FDG-PET accurately detected local disease spread and metastatic disease in patients with renal cell carcinoma and altered treatment in 40%. FDG-PET may have a role in the diagnostic evaluation of patients with renal cell carcinoma preoperatively and staging of metastatic disease. PMID- 11490228 TI - Complex perioperative immuno-dysfunction in patients with renal cell carcinoma. AB - PURPOSE: Patients with renal cell carcinoma have an impaired function of the immune system, which is the basis for different approaches of immunotherapy. We address perioperative changes of several parameters of the immune system in these patients. MATERIALS AND METHODS: Parameters of cellular and humoral immunity, including differential blood count, T cell markers CD2, 3, 4 and 8, B cell markers CD19 and 20, monocyte markers CD13 and 14, natural killer cell marker CD16, activation markers CD25, CD26 and HLA-DR, and cytokines interleukin-1 (IL 1) receptor antagonist, IL-2, soluble IL-2 receptor, IL-6, IL-10 and transforming growth factor-beta, were measured in the venous blood of patients who underwent renal surgery extracorporeal shock wave lithotripsy (ESWL, Dornier Medical Systems, Inc., Marietta, Georgia). Patients were grouped and age matched, and 37 underwent tumor nephrectomy, 20 open renal surgery for nonmalignant reasons and 24 ESWL. A group consisting of 39 controls received no treatment. RESULTS: Little change was detected in controls and those patients who received ESWL. Patients who underwent open renal surgery had increased leukocyte and granulocyte counts until postoperative day 3 but had low T cell counts. The postoperative decrease in CD25 expressing cells corresponded to an increase in the soluble IL-2 receptor. Cytokines IL-6 and 10, which also have immunosuppressive properties, were markedly increased postoperatively. These changes were more noted (p <0.01) in those patients who underwent tumor nephrectomy than open renal surgery for nonmalignant reasons and remained detectable when paired patients with similar surgical trauma were compared. In tumor nephrectomy cases renal venous IL-6 was higher than peripheral venous levels. CONCLUSIONS: Patients with renal cell carcinoma suffer from selective immuno-dysfunction, indicating a rationale for perioperative immunomodulation. PMID- 11490229 TI - Sexuality preserving cystectomy and neobladder: initial results. AB - PURPOSE: Standard cystectomy for bladder cancer in males and females includes removal of organs that are vital to normal sexual function. We report the initial results of modified cystectomy in males and females meant to preserve all sexual function, called sexuality preserving cystectomy and neobladder. MATERIALS AND METHODS: Sexuality preserving cystectomy and neobladder consists of pelvic lymph node dissection followed by cystectomy alone with preservation of the vasa deferentia, prostate and seminal vesicles in males, and all internal genitalia in females. An ileal neobladder is anastomosed to the margins of the prostate in males and urethra in females. Indications for this type of surgery are bladder cancer stages T1-T3 with absent tumor growth in the bladder neck in males and females, absent tumor in the prostatic urethra in males and absent invasive tumor in the trigone in females. Further requirements are patient motivation for the preservation of sexual function, no prostate cancer and no cervical/uterine abnormalities. Preoperative evaluation in males involves prostate specific antigen measurement and transrectal ultrasound with sextant prostate biopsies, while females undergo gynecological examination with a cervical smear and transvaginal ultrasound. Voiding and sexual function are assessed by a structured interview with preoperative urodynamics. Erectile function is evaluated by RigiScan (UroHealth Systems, Inc., Laguna Niguel, California) nocturnal penile erection measurement. A short course of 20 Gy. external radiation therapy to the bladder is given shortly before surgery. No patient has been lost to followup, which involves repeat RigiScan examination and regular endoscopy. All patients were entered in a prospective clinical trial approved by the medical ethics committee. RESULTS: From 1995 to 1998, 10 males and 3 females 38 to 71 years old (mean age 55) were enrolled in this protocol. Bladder cancer was stage T carcinoma in situ N0M0 in 1 case, Ta multiple grade 3 N0M0 in 1, T1 multiple grade 3 N0M0 in 4, T2 grade 3 N0M0 in 5, T2 grade 3 N1M0 in 1 and T3 grade 3 N1M0 in 1. Mean followup was 3.5 years (range 3 to 6). Two patients died of widespread metastasis without local recurrence. In 1 case prostate cancer developed 5 years after sexuality preserving cystectomy and neobladder, which was treated with external radiation therapy. Erection was normal in 7 men with antegrade ejaculation in 5 and vaginal lubrication was reported to be normal in all women. Daytime continence was achieved in 9 of the 10 males and 2 of the 3 females, while nighttime continence was achieved in 7 and 2, respectively. One woman and 3 men perform intermittent catheterization because of post-void residual urine after voiding. Postoperatively a vaginal fistula and ureteral stenosis developed in 1 case each. CONCLUSIONS: Sexuality preserving cystectomy and neobladder achieves maximal tissue conservation, resulting in preserved normal sexual function and satisfactory urinary tract reconstruction. Using strict criteria oncological results have not been jeopardized to date. PMID- 11490230 TI - Telomerase activity and cytokeratin 20 as markers for the detection and followup of transitional cell carcinoma: an unfulfilled promise. AB - PURPOSE: Telomerase activity compensates for the erosion of chromosomes and it has been detected in a wide variety of human tumors. Cytokeratin 20, an intermediate filament of epithelial cells, is expressed particularly in the urinary tract. These 2 molecules are candidates to become markers for the detection and followup of bladder carcinoma. We evaluate whether each molecule may serve as a potential marker and whether the 2 combined would improve the detection or followup of bladder carcinoma in a noninvasive manner. MATERIALS AND METHODS: We obtained 44 morning urine samples from patients with transitional cell carcinoma patients and 26 from age matched patients with a wide variety of clinical disorders but no malignancy of any kind. A telomerase polymerase chain reaction-enzyme-linked immunosorbent assay kit was used to determine telomerase activity and cytokeratin 20 expression was determined by nested reverse transcriptase-polymerase chain reaction. RESULTS: All samples tested positive for cytokeratin 8 expression, which verified epithelial cells in the urine samples. Of the 44 transitional cell carcinoma cases of all stages and grades 37 (84.1%) were positive for telomerase activity, 36 (81.8%) were positive for cytokeratin 20 expression and 65.9% were double positive. Of the 29 controls with various clinical conditions other that malignancy 22 (75.9%) were positive for telomerase activity, 13 (44.83%) were positive for cytokeratin 20 expression and 34.6% were double positive. CONCLUSIONS: Telomerase activity and cytokeratin 20 expression are not specific for malignancy and may be detected in many nonmalignant pathological conditions. Therefore, their use as potential markers of bladder carcinoma should be carefully reevaluated. PMID- 11490231 TI - The value of a single biopsy with 12 transperineal cores for detecting prostate cancer in patients with elevated prostate specific antigen. AB - PURPOSE: Prostate cancer detection on standard sextant biopsy is considered inadequate. Various biopsy protocols have been introduced to improve cancer diagnosis. We report our experience with transperineal 12-core prostate biopsy. MATERIALS AND METHODS: In a prospective study 650 patients underwent prostate specific antigen (PSA) measurement during a 15-month period, of whom 141 with PSA greater than 4 ng./ml. also underwent transperineal 12-core prostate biopsy using the fan technique. Median PSA was 8 ng./ml. (range 4.1 to 5,000). RESULTS: Prostate cancer was detected in 72 of the 141 patients (51%), including 44 of the 97 (45%) with PSA between 4.1 and 10 ng./ml. This incidence is higher than previously reported in the literature using other biopsy techniques. Disease was low grade Gleason 2 to 4 in 4 cases (5%), intermediate grade Gleason 5 to 6 in 26 (35%) and high grade Gleason 7 to 10 in the remaining 42 (60%). CONCLUSIONS: A high cancer detection rate is achieved by 12-core transperineal prostate biopsy. Most tumors represent clinically significant cancer. Further randomized trials are required to confirm these data. PMID- 11490232 TI - Prospective detection of clinically relevant prostate cancer in the prostate specific antigen range 1 to 3 ng./ml. combined with free-to-total ratio 20% or less: the Aarau experience. AB - PURPOSE: Little is known about the incidence rate and clinical relevance of prostate cancer in a low prostate specific antigen (PSA) level. In a prospective PSA based screening study we investigated the incidence and clinicopathological features of prostate cancer that occurred within PSA range 1 to 3 ng./ml. when the free-to-total ratio was 0.20 or less. MATERIALS AND METHODS: Men participating in the Aarau, Switzerland, section of the European Randomized Study of Screening for Prostate Cancer between October 1998 and July 2000 were included in the study. As a side study, all men with PSA between 1 and 3 ng./ml. and free to-total ratio 0.20 or less were invited to undergo further evaluation with ultrasound guided sextant prostate biopsy. RESULTS: Overall, 168 (7.8%) participants fulfilled inclusion criteria. A total of 158 (94%) patients underwent prostate biopsy, and prostate cancer was detected in 17 (10.8%). There were no statistically significant differences between prostate cancer and benign prostatic hyperplasia in regard to patient age (60.7 versus 59.8 years), prostate volume (23.9 versus 23.0 cc), PSA (1.98 versus 1.86 ng./ml.), free-to-total ratio (0.161 versus 0.160), PSA density (0.089 versus 0.076 ng./ml.) or PSA transition zone density (0.33 versus 0.24 ng./ml., respectively). Median Gleason score was 5 on prostate biopsy versus 6 on retropubic prostatectomy specimen. Of the 14 patients who underwent surgery there were positive lymph nodes in 1, stage pT3b Gleason 7 disease in 1, and pathologically organ confined Gleason 5 in 2, Gleason 6 in 5 and Gleason 7 in 5. Mean tumor volume was 1.01 cc (range 0.02 to 5.17). There were 2 (14.3%) insignificant (less than 0.2 cc, Gleason grade 3 or less), 1 (7.1%) minimal (less than 0.5cc, Gleason grade 3 or less) and 11 (78.6%) clinically relevant and potentially harmful cancers. CONCLUSIONS: There is a significant number of prostate cancer cases diagnosed at PSA as low as 1 to 3 ng./ml. A majority of these tumors are clinically significant. This free-to-total ratio range may be helpful for identifying prostate cancer. The "window of opportunity" for detection of curable cancer may change in populations with higher life expectancy towards lower PSA. Lack of specificity and characterization of tumor aggressiveness remains an unsolved issue for PSA. PMID- 11490233 TI - Safety and morbidity of first and repeat transrectal ultrasound guided prostate needle biopsies: results of a prospective European prostate cancer detection study. AB - PURPOSE: We prospectively evaluate the safety, morbidity and complication rates for first and repeat transrectal ultrasound guided prostate needle biopsies. MATERIALS AND METHODS: In this prospective European Prostate Cancer Detection Study 1,051 men, with total prostate specific antigen between 4 and 10 ng./ml., underwent transrectal ultrasound guided sextant biopsy plus 2 additional transition zone biopsies. Biopsy samples were also obtained from suspicious areas identified during transrectal ultrasound and digital rectal examination. All 820 patients with biopsy samples negative for prostate cancer underwent re-biopsy after 6 weeks. Immediate and delayed (range 1 to 7 days) morbidity, patient satisfaction and complication rates were recorded. RESULTS: Of the 1,051 subjects the initial biopsy was positive for prostate cancer in 231 and negative, including benign prostatic hyperplasia or benign tissue, in 820. Of these 820 patients prostate cancer was detected in 10% (83) on re-biopsy. Minor or no discomfort was observed in 92% and 89% of patients at first and re-biopsy, respectively (p = 0.29). Immediate morbidity was minor and included rectal bleeding (2.1% versus 2.4%, p = 0.13), mild hematuria (62% versus 57%, p = 0.06), severe hematuria (0.7% versus 0.5%, p = 0.09) and moderate to severe vasovagal episodes (2.8% versus 1.4%, respectively, p = 0.03). Delayed morbidity of first and re-biopsy was comprised of fever (2.9% versus 2.3%, p = 0.08), hematospermia (9.8% versus 10.2%, p = 0.1), recurrent mild hematuria (15.9% versus 16.6%, p = 0.06), persistent dysuria (7.2% versus 6.8%, p = 0.12) and urinary tract infection (10.9% versus 11.3%, respectively, p = 0.07). Major complications were rare and included urosepsis (0.1% versus 0%) and rectal bleeding that required intervention (0% versus 0.1%, respectively). Furthermore, an age dependent pattern of pain apprehension during biopsy was observed with the highest scores in patients younger than 60 years. CONCLUSIONS: Transrectal ultrasound guided biopsy is generally well tolerated with minor morbidity only rarely requiring treatment. Re-biopsy can be performed 6 weeks later with no significant difference in pain or morbidity. Patients younger than 60 years should be counseled in regard to a higher level of discomfort, and local and topical anesthesia if desired. PMID- 11490234 TI - Interval after prostate specific antigen testing and subsequent risk of incurable prostate cancer. AB - PURPOSE: Studies of the potential effect of prostate specific antigen (PSA) screening on a less than yearly basis have been limited to computer simulations using relatively small sample sets. Primary clinical data on this relationship have not been generally available. We examined the relationship of less frequent testing and the risk of nonlocalized incurable cancer. The effect of testing frequency on the risk of prostate biopsy in men ultimately diagnosed with cancer was also assessed. MATERIALS AND METHODS: The study included a population based sample of 36,422 men 65 years old or older residing in 9 geographic areas with newly diagnosed prostate cancer during 1989 to 1993. The primary end point was the risk of nonlocalized cancer, as determined by logistic regression. Patient age, geographic region, year of diagnosis and race were included as covariates. RESULTS: In men who would be diagnosed with prostate cancer the risk of nonlocalized cancer did not differ in those tested 2 or 3 years compared with the risk in those tested 1 year before diagnosis (relative risk 1.00, 95% confidence interval 0.84 to 1.20 and 1.02, 95% confidence interval 0.74 to 1.41, respectively). However, the risk of prostate biopsy in these men was directly related to the number of PSA tests performed (test for trend p = 0.0061). CONCLUSIONS: Patients who choose to undergo PSA testing may be tested on a biennial instead of annual basis without an increased risks of nonlocalized cancer. Decreasing the frequency of PSA testing may lead to fewer prostate biopsies. PMID- 11490235 TI - Repeat biopsy strategy in patients with atypical small acinar proliferation or high grade prostatic intraepithelial neoplasia on initial prostate needle biopsy. AB - PURPOSE: Isolated high grade prostatic intraepithelial neoplasia and/or atypical small acinar proliferation on prostate biopsy increases the risk of identifying cancer on repeat biopsy. We report the results of repeat prostate biopsy for high grade prostatic intraepithelial neoplasia and/or atypical small acinar proliferation, and propose an optimal repeat biopsy strategy. MATERIALS AND METHODS: Of 1,391 men who underwent standard systematic sextant biopsy of the prostate 137 (9.8%) had isolated high grade prostatic intraepithelial neoplasia or atypical small acinar proliferation, including 100 who underwent repeat prostate biopsy within 12 months of the initial biopsy. RESULTS: Adenocarcinoma was detected in 47 of the 100 patients who underwent repeat biopsy. The initial biopsy site of high grade prostatic intraepithelial neoplasia and/or atypical small acinar proliferation matched the sextant location of cancer on repeat biopsy in 22 cases (47%). Repeat biopsy directed only to the high grade prostatic intraepithelial neoplasia and/or atypical small acinar proliferation site on initial biopsy would have missed 53% of cancer cases. In 12 of the 47 men (26%) cancer was limited to the side of the prostate contralateral to the side of high grade prostatic intraepithelial neoplasia and/or atypical small acinar proliferation. Of the 31 patients with cancer in whom the transition zone was sampled cancer was limited to the transition zone in 4 (13%) and evident at other biopsy sites in 13 (42%). The only significant predictor of positive repeat biopsy was mean prostate specific antigen velocity plus or minus standard error (1.37 +/- 1.4 versus 0.52 +/- 0.8 ng./ml. per year, p <0.001). CONCLUSIONS: Patients with isolated high grade prostatic intraepithelial neoplasia and/or atypical small acinar proliferation on prostate biopsy are at 47% risk for cancer on repeat biopsy. The optimal repeat biopsy strategy in this setting should include bilateral biopsies of the standard sextant locations. We also strongly recommend that transition zone sampling should be considered. PMID- 11490236 TI - The expression of neuropeptides in hyperplastic and malignant prostate tissue and its possible clinical implications. AB - PURPOSE: We characterized the incidence and pattern of distribution of neuroendocrine differentiated tumor cells in prostatic hyperplastic and carcinomatous tissue, correlated neuroendocrine differentiation with prostate specific antigen (PSA) and assessed whether neuroendocrine cells have value as an independent indicator of poor prognosis in patients with prostate carcinoma. MATERIALS AND METHODS: We immunohistochemically evaluated hyperplastic and carcinomatous prostate specimens for chromogranin A, neuron specific enolase and serotonin expressing tumor cells. The expression of various markers in cells was analyzed and correlated with tumor DNA ploidy, disease grade and stage, PSA and clinical course in patients with prostate cancer. RESULTS: Enrolled in our study were 31 patients with hyperplastic prostate tissue and 30 with prostatic carcinoma. Followup in cancer cases was 1 to 9 years (mean 3.7). During followup 9 patients (30%) died of cancer. We noted DNA content aneuploidy in 5 cases (16.7%) of prostate carcinoma. Chromogranin A, neuron specific enolase and serotonin were expressed in 80%, 43% and 77% of cases of prostate carcinoma and in 29%, 10% and 36% of hyperplastic tissue, respectively. Larger prostates had no higher content of various neuroendocrine cells than smaller prostates. There was higher expression of neuropeptides in carcinomatous than in hyperplastic tissue. Of the 3 peptides chromogranin A was significantly related to all parameters, including Gleason score, tumor stage, PSA and patient survival. In addition to PSA, neuron specific enolase was also closely associated with other clinicopathological parameters. Serotonin was significantly related to patient survival only but we noted no correlation with Gleason score, tumor stage or PSA. In regard to factors predictive of patient prognosis expression of the 3 neuropeptides in tumor cells, Gleason score, tumor stage and PSA were closely related to patient survival in this study CONCLUSIONS: The growth of hyperplastic prostate tissue is related to neuroendocrine cell activity. The chromogranin A marker has the highest expression in prostate cancer. Neuroendocrine cells may represent an independent indicator of poor prognosis in patients with prostate carcinoma. PMID- 11490237 TI - High dose radiation delivered by intensity modulated conformal radiotherapy improves the outcome of localized prostate cancer. AB - PURPOSE: We present the long-term outcome and tolerance of 3-dimensional (D) conformal and intensity modulated radiation therapy for localized prostate cancer. MATERIALS AND METHODS: Between October 1988 and December 1998, 1,100 patients with clinical stages T1c-T3 prostate cancer were treated with 3-D conformal or intensity modulated radiation therapy. Patients were categorized into prognostic risk groups based on pretreatment prostate specific antigen (PSA), Gleason score and clinical stage. Sextant biopsies were performed 2.5 years or greater after treatment to assess local control. PSA relapse was defined according to the consensus guidelines of the American Society for Therapeutic Radiation Oncology. Late toxicity was classified according to the Radiation Therapy Oncology Group morbidity grading scale. Median followup was 60 months. RESULTS: At 5 years the PSA relapse-free survival rate in patients at favorable, intermediate and unfavorable risk was 85% (95% confidence interval [CI] +/- 4), 58% (95% CI +/- 6) and 38% (95% CI +/- 6), respectively (p <0.001). Radiation dose was the most powerful variable impacting PSA relapse-free survival in each prognostic risk group. The 5-year actuarial PSA relapse-free survival rate for patients at favorable risk who received 64.8 to 70.2 Gy. was 77% (95% CI +/- 8) compared to 90% (95% CI +/- 8) for those treated with 75.6 to 86.4 Gy. (p = 0.04) [corrected]. The corresponding rates were 50% (95% CI +/- 8) versus 70% (95% CI +/- 6) in intermediate risk cases (p = 0.001), and 21% (95% CI +/- 8) versus 47% (95% CI +/- 6) in unfavorable risk cases (p = 0.008) [corrected]. Only 4 of 41 patients (10%) who received 81 Gy. had a positive biopsy 2.5 years or greater after treatment compared with 27 of 119 (23%) after 75.6, 23 of 68 (34%) after 70.2 and 13 of 24 (54%) after 64.8 Gy. The incidence of toxicity after 3-D conformal radiation therapy was dose dependent. The 5-year actuarial rate of grade 2 rectal toxicity in patients who received 75.6 Gy. or greater was 14% (95% CI +/- 2) compared with 5% (95% CI +/- 2) in those treated at lower dose levels (p <0.001). Treatment with intensity modulated radiation therapy significantly decreased the incidence of late grade 2 rectal toxicity since the 3-year actuarial incidence in 189 cases managed by 81 Gy. was 2% (95% CI +/- 2) compared with 14% (95% CI +/- 2) in 61 managed by the same dose of 3-D conformal radiation therapy (p = 0.005). The 5-year actuarial rate of grade 2 urinary toxicity in patients who received 75.6 Gy. or greater 3-D conformal radiation therapy was 13% compared with 4% in those treated up to lower doses (p <0.001). Intensity modulated radiation therapy did not affect the incidence of urinary toxicity. CONCLUSIONS: Sophisticated conformal radiotherapy techniques with high dose 3-D conformal and intensity modulated radiation therapy improve the biochemical outcome in patients with favorable, intermediate and unfavorable risk prostate cancer. Intensity modulated radiation therapy is associated with minimal rectal and bladder toxicity, and, hence, represents the treatment delivery approach with the most favorable risk-to-benefit ratio. PMID- 11490238 TI - Safety and efficacy of exisulind for treatment of recurrent prostate cancer after radical prostatectomy. AB - PURPOSE: We evaluated the safety and efficacy of exisulind for delaying disease progression in men with increasing prostate specific antigen (PSA) after radical prostatectomy. MATERIALS AND METHODS: A total of 96 men with increasing PSA after radical prostatectomy were randomized to receive placebo (49) or 250 mg. exisulind twice daily (47) for 12 months. The primary efficacy parameter was the difference in change from baseline PSA between the placebo and exisulind groups. The PSA doubling time was also evaluated before and during study. A subgroup analysis classified patients based on the risk of developing metastatic disease. RESULTS: Compared with placebo, exisulind significantly suppressed the increase in PSA in all patients (p = 0.017). The results were also statistically significant in men at high risk for metastasis (p = 0.0003) and those who could not be classified according to risk (p = 0.0009). In addition, median PSA doubling time was lengthened in high risk patients on exisulind (2.12 month increase) compared with those on placebo (3.37 month decrease, p = 0.048). Exisulind was well tolerated. CONCLUSIONS: Exisulind inhibited the increase in PSA overall and prolonged PSA doubling time in high risk patients compared with placebo. These results suggest that Exisulind has the potential to extend the time from biochemical recurrence to the need for androgen deprivation therapy. Exisulind was well tolerated in this patient population. Our results support further study of Exisulind in the treatment of patients with prostate cancer. PMID- 11490239 TI - Glanuloplasty with scrotal flap for partial penectomy. AB - PURPOSE: Reconstructing a penile stump secondary to trauma or cancer should result in satisfactory penile function and appearance. The lack of penile skin, stump retraction in the scrotum and stenosis of the neomeatus must be resolved in these cases. MATERIALS AND METHODS: A 2-stage surgical technique with a scrotal flap was used in 34 patients with a mean age of 43.2 years to reconstruct the glans. Mean followup was 73.2 months. After penectomy a scrotal flap was designed and its distal extreme was transferred to the penile stump. The urethral end was sutured to a hole in the scrotal flap and the flap borders were sutured to the adjacent albuginea. The flap pedicle was resected 4 to 6 weeks later. RESULTS: Patient recovery was characterized by a normal-appearing penis and unobstructed urinary flow. Definite depilation of the neoglans was required in 17.6% of cases. Partial necrosis of 2 flaps (5.8%) required grafts. Sexual potency was preserved in 7 men (20.5%). In 1 case (2.9%) urethral meatal stenosis resolved with minor surgical procedures. CONCLUSIONS: This technique enables us to design a neoglans with acceptable function and appearance, no penile retraction, satisfactory voiding and in certain cases possible intercourse with vaginal penetration. PMID- 11490240 TI - Optimal prevention and management of proximal ureteral stent migration and remigration. AB - PURPOSE: We evaluated issues associated with proximal ureteral stent migration and remigration, including causes and management, and the predictability of ureteral length. MATERIALS AND METHODS: All proximal ureteral stent migrations that occurred from January 1997 to March 2000 were reviewed. Characteristics and treatment of the 33 patients with proximal ureteral stent migration were compared with those of 66 randomly selected controls who did not have stent migration. We also analyzed a subgroup of 6 cases of remigration. RESULTS: Of the ureteral stents 2% migrated proximally. Mean height was greater in patients with versus without a migrated stent (p = 0.028). The stent-to-ureter length ratio was lower in the migrated than in the nonmigrated group (p <0.0001). Patient height and side of migration were significant predictors of ureteral length (R2 = 0.3511, p <0.0001 and 0.0007, respectively). Of the patients who required continued ureteral stenting migrated stent management included placement of a longer stent in 9 (group 1) and a stent of equal length in 4 (group 2), and repositioning of the original stent in 4 (group 3). There was no remigration in group 1. However, migration recurred in 2 patients in group 2 (50%) and in all 4 in group 3 (100%). CONCLUSIONS: Proximal migration occurs when a stent is too short for the ureter. We recommend that ureteral length should be measured directly from an x-ray to select the optimal stent length. If it is necessary to continue stenting a ureter after migration has been detected, a longer stent should be placed. PMID- 11490241 TI - Periprostatic nerve blockade for transrectal ultrasound guided biopsy of the prostate: a randomized, double-blind, placebo controlled study. AB - PURPOSE: We performed a randomized, double-blind, placebo controlled study to assess the safety and efficacy of periprostatic anesthesia administration during prostate biopsy. MATERIALS AND METHODS: From May to November 2000 transrectal ultrasound guided prostate biopsy was performed in 132 consecutive men due to an abnormal digital rectal examination and/or elevated prostate specific antigen. During biopsy 66 patients each were randomly assigned to receive an injection of 1% lidocaine or normal saline. Immediately after biopsy the pain score was recorded independently by patients and the physician using a 10-point linear scale. In addition, patients were given a descriptive questionnaire to be completed at home and mailed back within 2 weeks of biopsy. RESULTS: Mean patient perceived pain scores plus or minus standard deviation of 2.7 +/- 0.21 in the lidocaine and 4.7 +/- 0.26 in the saline groups were significantly different (p <0.0001), as were mean physician perceived scores of 2.9 +/- 0.27 and 5.1 +/- 0.34, respectively (p = 0.0001). Mean questionnaire scores for pain during biopsy of 2.3 +/- 0.13 in the lidocaine and 3.1 +/- 0.18 in the saline groups were also significantly different (p = 0.0006), as were mean questionnaire scores for pain after biopsy of 1.8 +/- 0.11 and 2.3 +/- 0.13, respectively (p <0.006). There were no adverse effects of injection. CONCLUSIONS: Our results show a significant benefit of periprostatic anesthesia over placebo in a randomized double-blind trial. This safe, simple and rapid technique should be applied at transrectal ultrasound guided prostate biopsy to limit undue patient discomfort. PMID- 11490242 TI - A nonrefluxing, serous lined extramural tunnel for ureteroileal anastomosis in ileal conduit urinary diversion: first clinical experience in 10 patients. AB - PURPOSE: We report the surgical technique and functional outcome of a new application for serous lined, anti-refluxing ureteroileal anastomosis in ileal conduit urinary diversion. Reflux prevention relies on the construction of a serous lined extramural ileal tunnel. MATERIALS AND METHODS: A 25 cm. distal ileal segment was isolated. The proximal 7 cm. of the ileal segment was folded and the 2 proximal 7 cm. segments were joined by seromuscular sutures. The antimesenteric borders of these 7 cm. segments were incised and the medial edges of each ureter were joined. A mesenteric window was opened at the level of ileal folding and the ureters were passed through it. They were inlaid within the trough and the conjoined ureteral end were anastomosed to the intestinal mucosa. The tunnel was then closed over the implanted ureters. The lateral limbs of the detubularized ileal segment were then joined. The technique was performed in 10 patients with a mean followup of 9.9 months (range 3 to 19). The patients were evaluated clinically and radiologically. RESULTS: None of the 10 patients had reflux on x-ray of the loop. One patient had previously undergone unilateral nephrectomy. Excretory urography showed a stabilized or improved upper tract in 18 renal units. Left ureterohydronephrosis was present in 1 renal unit because of ureteroileal stenosis. CONCLUSIONS: The initial clinical results of the serous lined extramural ileal tunnel technique for ureteroileal anastomosis in ileal conduit cases are promising. The technique appears effective and reliable. PMID- 11490243 TI - Influence of stent size on the success of antegrade endopyelotomy for primary ureteropelvic junction obstruction: results of 2 consecutive series. AB - PURPOSE: We evaluated the influence of stent size in 2 consecutive series of unselected patients in whom primary ureteropelvic junction obstruction was managed by antegrade endopyelotomy and stenting with a 14 or 27Fr stent at the level of the incision. MATERIALS AND METHODS: Antegrade endopyelotomy was performed in 132 patients with primary ureteropelvic junction obstruction. The endopyelotomy was stented for 6 weeks. In 77 patients (group 1) a 14/8.2Fr percutaneous endopyelotomy (Smith) catheter was used. In 55 patients (group 2) a modified 14/8.2Fr Smith catheter was over pulled with a 27Fr wound drain. The wound drain was removed after 2 to 3 weeks and the standard 14/8.2Fr stent remained in place for another 3 to 4 weeks. Success at 6 to 8 weeks, and 6 and 24 months postoperatively was based on clinical evaluation, and excretory urography and/or diuretic renography. Thereafter clinical and ultrasound followup was performed every 2 to 3 years. RESULTS: Preoperatively data on the risk factors of large pyelocaliceal volume and impaired renal function were similar in the 2 groups. The overall success rate was 70% in group 1 at a median followup of 67 months (range 2 to 118) and 94% in group 2 at a median followup of 23 months (range 2 to 52). The early success rate after 6 to 8 weeks in groups 1 and 2 was 83% and 94%, respectively. The long-term success rate after 2 years was 71% and 93%, respectively. Perioperatively and postoperatively the incidence of complications was 16% in group 1 and 24% in group 2. When group 2 complications due to a lack of experience with the new stent were excluded from analysis, the remaining 15% complication rate was comparable to that in group 1. Mean pyelocaliceal volume decreased significantly in each group and remained stable. Split renal function did not change preoperatively to postoperatively with no significant difference in the 2 groups. CONCLUSIONS: Stenting an antegrade endopyelotomy with a modified 27Fr instead of a 14Fr catheter seems to increase the early and, even more impressively, the long-term success rate to a level similar to that of open pyeloplasty. PMID- 11490244 TI - Videourodynamic studies in men with lower urinary tract symptoms: a comparison of community based versus referral urological practices. AB - PURPOSE: We compared the clinical and urodynamic characteristics of men referred for evaluation of lower urinary tract symptoms in community based versus referral urological practices and examined the various pathophysiological mechanisms of these symptoms. MATERIALS AND METHODS: We reviewed a multicenter urodynamics database of 963 consecutive men referred for the evaluation of persistent lower urinary tract symptoms at 2 community based and 1 urological referral center. Of the 963 patients in the database 422 (44%) were excluded from study due to neurological disorder in 41%, previous urinary or pelvic surgery in 27% and the use of medications known to affect voiding in 24%. A total of 541 patients with a mean age plus or minus standard deviation of 64.4 +/- 13.8 years met study inclusion criteria and were analyzed further. We compared the clinical and urodynamic characteristics of patients at the community and referral centers. RESULTS: Lower urinary tract symptoms were equally common in men presenting to community and referral centers. The most common symptom was difficult voiding, followed by frequency, urgency and nocturia in 58%, 54%, 43% and 40% of the study population, respectively. Urodynamic diagnoses were also similar in the 2 groups. Although bladder outlet obstruction was diagnosed in 69% of patients, it was the only urodynamic finding in a third of the patients with obstruction. The main concomitant urodynamic diagnoses were detrusor overactivity, bladder hyposensitivity, impaired detrusor contractility, low bladder compliance and bladder hypersensitivity in 47%, 10%, 10%, 9% and 3% of obstructed cases, respectively. CONCLUSIONS: The pathophysiology of lower urinary tract symptoms in men is multifactorial, and similar at community practice and tertiary referral centers. The disparity in urodynamic findings and subjective symptoms emphasizes the need for a thorough and early clinical and urodynamic evaluation. PMID- 11490245 TI - Posterior tibial nerve stimulation as neuromodulative treatment of lower urinary tract dysfunction. AB - PURPOSE: Recently, intermittent percutaneous posterior tibial nerve stimulation was introduced as a treatment modality filling the gap between conservative and surgical therapies in patients with certain types of lower urinary tract dysfunction. MATERIALS AND METHODS: In a prospective multicenter trial posterior tibial nerve stimulation was evaluated in 37 patients who presented with symptoms of bladder overactivity, that is the urgency and frequency syndrome and/or urge incontinence, and 12 with nonobstructive urinary retention. Results were recorded in voiding diaries and on quality of life questionnaires before and after treatment. Patients were classified as responders, including those in whom therapy was successful and chose to continue treatment after the initial 12 weeks, and nonresponders, those who chose to stop treatment. RESULTS: Overall, a positive response was seen in 60% of all patients. In patients with bladder overactivity a statistically significant decrease was observed in leakage episodes, number of pads used, voiding frequency and nocturia, and an equal increase in mean and smallest volume voided. Improvements were also seen in nonobstructive urinary retention, including number of catheterizations, total and mean volume catheterized, and total and mean volume voided. Disease specific quality of life and some domains of general quality of life improved, especially of bladder overactivity. Only mild side effects were observed. CONCLUSIONS: Posterior tibial nerve stimulation is a minimally invasive and successful treatment option for patients with certain types of lower urinary tract dysfunction. PMID- 11490246 TI - Expansion and bioabsorption of the self-reinforced lactic and glycolic acid copolymer prostatic spiral stent. AB - PURPOSE: Self-reinforced bioabsorbable stents can be made self-expanding due to the viscoelastic memory of the oriented bioabsorbable materials. A new self expandable self-reinforced copolymer of lactic/glycolic acid, lactic/glycolic molar ratio 80:20 stent was developed to prevent postoperative urinary retention after procedures that induced prostatic edema. In in vitro experiments the expansion rate has been up to 100% during the first few hours at body temperature. We investigated the expansion rate and biodegradation of the self reinforced lactic and glycolic acid copolymer prostatic spiral stent in vivo in the prostatic urethra. MATERIALS AND METHODS: A total of 39 men, 52 to 84 years old, with lower urinary tract symptoms due to benign prostatic enlargement underwent interstitial laser coagulation of the prostate. A self-reinforced copolymer of lactic/glycolic acid, lactic/glycolic molar ratio 80/20 stent was inserted into the prostatic urethra at the end of the operation. The stent lumen diameter was 4.5 mm. The location and diameter of the lumen and degradation of the stent were studied with transrectal ultrasound at 1, 2, 4 and 6 months postoperatively. At 6 months patients underwent cystoscopy. RESULTS: All except 1 patient voided on postoperative day 1. Mean lumen diameter was 7.4 mm. (range 6.2 to 8.2) at 1 month and 7.2 mm (range 6.2 to 7.5) at 2 months. At 4 months the stent was degraded into small pieces. No pieces of stent were found in the prostatic urethra on ultrasound or cystoscopy at 6 months. However, a portion of the spiral stent was found at the bottom of the bladder in 2 patients. CONCLUSIONS: The speed and expansion rate of the self-reinforced copolymer of lactic/glycolic acid, lactic/glycolic molar ratio 80/20 stent was sufficient to lock the stent in place and ensure voiding in cases of edema induced bladder outlet obstruction. Strength retention greater than 2 months was long enough to avoid later impairments of voiding. PMID- 11490247 TI - Improving the accuracy of vascular testing in impotent men: correcting hemodynamic alterations using a vasoactive medication re-dosing schedule. AB - PURPOSE: Arteriogenic or venogenic vascular insufficiency is the most common pathophysiology of organic erectile dysfunction. While vascular insufficiency may be suspected on history and physical examination, the definitive diagnosis is made by vascular testing. Dynamic infusion cavernosometry is a recognized technique for evaluating the hemodynamics of erection. Assigning the correct vascular diagnosis during testing is important. Complete corporeal smooth muscle relaxation is essential for obtaining accurate data. Previously others have suggested that repeat dosing with vasoactive medication may improve the diagnostic accuracy of vascular testing. We investigated the hemodynamic effect of repeat doses of medication. MATERIALS AND METHODS: Data were prospectively obtained on men undergoing dynamic infusion cavernosometry. When veno-occlusive parameters were abnormal, the intracavernous vasoactive agent dose was repeated to a maximum of 3 doses. Standard dynamic infusion cavernosometry criteria were used for diagnosing arteriogenic and venogenic erectile dysfunction. We analyzed the proportion of men in whom the vascular diagnosis was altered using this regimen. RESULTS: Of 420 men undergoing dynamic infusion cavernosometry, 70% warranted re-dosing based on abnormal veno-occlusive parameters. Of these 294 men veno-occlusive values were corrected by repeat medication in 32% and, therefore, they would have been falsely diagnosed with venous leakage during vascular evaluation. Half of all corrections occurred with the second dose, while half of the patients required a third dose of medication. CONCLUSIONS: These data should encourage clinicians to consider re-dosing during the vascular evaluation of men in whom incomplete corporeal smooth muscle relaxation is suspected. In this way a false diagnosis of venous leakage may be avoided in a significant number of cases. PMID- 11490248 TI - Long-term efficacy of sildenafil and tachyphylaxis effect. AB - PURPOSE: We observed that patients who initially responded to sildenafil frequently became resistant to it with time. We evaluated the long-term efficacy of sildenafil. MATERIALS AND METHODS: A telephone survey was conducted of patients during the first year of sildenafil usage, and another one was completed 2 years later of the same group. RESULTS: During the first survey, the etiology of impotence included post radical prostatectomy in 25, arterial insufficiency in 26, diabetes in 19, neurogenic impotence in 12, suspected venous leak in 9, proved venous leak in 7, Peyronie's disease in 6 and unspecified in 47 patients. The overall improvement rate, which was defined as the ability to initiate and maintain erections for successful intercourse, was 74%. The dose necessary to achieve this response was 100 mg. sildenafil in 15% of patients, 50 mg. in 83% and 25 mg. in 2%. During the second survey, information was collected on 82 patients and only 43 (52%) had continued treatment. Of the 69 patients who reported an initial good response 41 (59%) were still using sildenafil, and of the 43 who were still using it 16 (37%) had to increase the dose by 50 mg. to achieve an adequate result. There was no significant relationship between the need to increase the dose and frequency of treatment per month. Reduction in efficacy ranged from 15% to 50% (mean 36 +/- 12%) and the time to loss of efficacy ranged from 1 to 18 months (mean 11 +/- 5). Of the 82 patients in the second surgery, 39 (48%) stopped using sildenafil. A total of 28 (74%) patients reported a good initial response on the first survey, and 6 had spontaneous erections and no longer needed treatment. However, 14 (50%) patients discontinued sildenafil because of the loss of efficacy. In this group 10 patients had side effects, mainly headache and flushing of the face and nose, and only 2 discontinued treatment because of them. CONCLUSIONS: Our data suggest that there is a possible tachyphylaxis effect with sildenafil. Of the patients who were followed for 2 years 20% had to increase the sildenafil dose to have the same effect and 17% discontinued use due to loss of efficacy. PMID- 11490249 TI - Mechanical reliability and safety of, and patient satisfaction with the Ambicor inflatable penile prosthesis: results of a 2 center study. AB - PURPOSE: We reviewed the experience with the 2-piece Ambicor penile prosthesis (American Medical Systems, Minnetonka, Minnesota) at 2 medical centers to investigate its mechanical reliability and complication rates as well as patient and partner satisfaction with the device. MATERIALS AND METHODS: From 1995 through 1999, 131 men underwent implantation of an Ambicor penile prosthesis at 2 medical centers. We performed a 3-part study consisting of a retrospective clinical record review, mailed patient and partner questionnaire, and mailed modified patient and partner Erectile Dysfunction Inventory of Treatment Satisfaction questionnaire. Mean followup was 43.4 months (range 12 to 73). RESULTS: All 131 men had a history consistent with an organic etiology of erectile dysfunction, including vascular disease in 62%, radical retropubic prostatectomy in 17%, Peyronie's disease in 15%, neophallus construction in 4% and radical pelvic surgery in 2%. Mean patient age was 56.8 years (range 22 to 76) at implantation. Overall there were complications in 10 cases (7.6%), including infection, hematoma and mechanical failure in 6 (4.6%), 1 (0.7%) and 3 (2.3%), respectively. A total of 112 men (85%) and 91 partners completed the questionnaire. All patient respondents still had an Ambicor prosthesis implanted and 96.4% had erection suitable for coitus. Overall patient and partner satisfaction was 96.4% and 91.2%, respectively. Of the respondents 92.9% of patients and 90.1% of partners would recommend the device to others. Of the 85 men (65%) and 46 partners who completed the modified Erectile Dysfunction Inventory of Treatment Satisfaction survey 90.6% and 82.6%, respectively, were satisfied or very satisfied overall with the penile prosthesis. CONCLUSIONS: The Ambicor penile prosthesis is associated with a low complication rate and reliable mechanical function. High satisfaction was reported by patients and partners. PMID- 11490250 TI - Radical cystectomy is safe in elderly patients at high risk. AB - PURPOSE: Radical cystectomy is standard treatment for bladder cancer in healthy individuals. We determined the safety of radical cystectomy in elderly patients at high risk. MATERIALS AND METHODS: We reviewed the records of all patients who underwent radical cystectomy at our institution between January 1994 and June 2000. Of these 382 patients we identified 44 who were elderly and at high risk, as defined by age 75 years or greater and American Society of Anesthesiologists classification 3 or greater. We examined postoperative care, perioperative minor/major complications, the mortality rate and the need for rehospitalization. RESULTS: Median age of the 44 patients was 77.5 years (range 75 to 87). American Society of Anesthesiologists class was 3 in 40 patients and 4 in 4. Median hospitalization was 7 days (range 4 to 20). Postoperatively 31 of the 44 patients (70%) were transferred directly to the general urology floor, while cardiac monitoring was required postoperatively in 30%. Nine of these patients were transferred to a step-down unit and the remaining 4 required surgical intensive care unit admission. Minor and major complications developed in 10 (22.7%) and 2 (4.5%) cases, respectively. No patients died in the perioperative period and 4 patients were hospitalized within 6 months of discharge home. CONCLUSIONS: Our results support the safety of radical cystectomy in elderly patients at high risk. Acceptable perioperative morbidity and mortality may be achieved without routine intensive monitoring postoperatively. PMID- 11490251 TI - Predictors of utilities for health states in early stage prostate cancer. AB - PURPOSE: When faced with treatment choices for early stage prostate cancer, patients must balance the survival benefit of a treatment with its morbidity. Little is known about how patients balance these trade-offs. To further our understanding of patient decision making we assessed patient utilities for prostate cancer treatment related morbidities. We determined whether patient utilities were predicted by sociodemographic characteristics or baseline genitourinary function. MATERIALS AND METHODS: We evaluated 401 men undergoing prostate needle biopsy for suspicion of prostate cancer at university, Veterans Affairs and public hospitals. Study design included a prospective cross-sectional cohort with correlation and multivariate analysis. Subjects were studied with 2 established health related quality of life instruments. Patient utilities were assessed with an interactive software application. RESULTS: On multivariate analysis utility for current general health was a significant predictor of utilities for treatment related morbidities. Surprisingly baseline urinary, sexual and bowel function scores did not correlate well with respective utilities for potential incontinence, impotence or radiation proctitis. In other words, men with good and imperfect baseline function were equally willing to risk impairment to preserve life. CONCLUSIONS: Men who perceived that general health was better appear to place higher value on quantity of life, while those who already are suffering from poor general health place higher value on quality of life. Ethnicity appears to modify some effects of other variables on patient preference. Utility assessment provides a quantitative tool to aid physicians in counseling patients when making treatment decisions for localized prostate cancer. PMID- 11490252 TI - Quality of life after treatment for localized prostate cancer: differences based on treatment modality. AB - PURPOSE: Brachytherapy with 103palladium (103Pd) is an increasingly administered treatment modality for localized prostate cancer. We compared general and disease specific health related quality of life after 103Pd treatment, radical prostatectomy and external beam radiation therapy given during the same time frame. MATERIALS AND METHODS: We performed a retrospective cross-sectional survey study of patients treated at a single community medical center between 1995 and 1999. We mailed 5 validated health related quality of life survey instruments to 269, 142 and 222 men who underwent radical prostatectomy, 103Pd treatment and external beam radiation therapy, respectively, with a response rate of greater than 80% in all groups. RESULTS: General health related quality of life assessed by the SF-36 showed the same scores in patients who underwent prostatectomy and 103Pd treatment. The University of California-Los Angeles Prostate Cancer Index was used to assess bowel, urinary and sexual function/bothersomeness. External beam radiation therapy reported was associated with worse bowel function and greater bowel bothersomeness. Prostatectomy was associated with worse urinary function compared to 103Pd and external beam radiation therapy. Prostatectomy was associated with worse sexual function than 103Pd or external beam radiation therapy, although nerve sparing surgery and erectile aids minimized the difference. American Urological Association symptom scores were initially higher for 103Pd but became equal to those in the other groups in patients treated greater than 12 months from survey time. Disease-free men who underwent prostatectomy and 103Pd brachytherapy were equally confident that cancer would not recur in the future. Satisfaction rates were equivalent and biochemical failure significantly decreased satisfaction in all groups. CONCLUSIONS: While general health related quality of life was mostly unaffected by the 3 most common treatments for prostate cancer, there were differences in bowel, urinary and sexual function. This information may aid patients in the decision making process. PMID- 11490253 TI - Patient reported complications after prostate brachytherapy. AB - PURPOSE: Prostate brachytherapy has gained popularity due partly to the low rates of short-term complications shown in studies from highly select clinical practices. These series rely on medical records generated by the treating physician and are prone to underreport complications. We summarize the complication reports obtained directly from patients to establish a more realistic incidence of treatment related problems. MATERIALS AND METHODS: In 1997, 160 consecutive patients treated with prostate brachytherapy at the University of Washington were studied. A questionnaire was designed to determine the rate of complications occurring within 1 year of the procedure. The questions were formulated for ease of use and conciseness, while accounting for easily recalled events associated with complications. A total of 147 (92%) patients completed the questionnaire. RESULTS: There were 8 (5%) patients who required hospital admission for an average of 2 days (range 1 to 7) as a result of the procedure. A total of 56 (38%) patients required nonroutine visits with a physician in an office setting or at an emergency room. Radiation proctitis diagnosed by endoscopy developed in 8 (5%) patients but no one needed surgical intervention. A total of 47 (32%) patients required urinary catheterization at some point after implantation. CONCLUSIONS: We demonstrated a higher rate of short-term complications than those previously reported. Fortunately, the majority of side effects were self-limited and no treatment related mortality or cardiovascular morbidity was seen. Our findings may provide a more realistic account of the complications likely to occur after implantation than might be surmised from previous reports. PMID- 11490254 TI - Assessment of early continence recovery after radical prostatectomy: patient reported symptoms and impairment. AB - PURPOSE: Patients considering radical prostatectomy often inquire as to when they can expect to regain urinary continence. However, there is a paucity of patient self-reported data regarding the recovery of continence during the initial 3 months after surgery. Our objectives were to assess urinary continence changes early in the postoperative period and determine which of 2 commonly used definitions of continence more closely relate to patient reported urinary impairment. MATERIALS AND METHODS: A prospective study of 90 men with clinically localized prostate cancer who selected radical prostatectomy as primary therapy was conducted. Repeated measures of urinary continence as defined by 1) total urinary control, 2) the use of 1 or 0 pads daily, and 3) small or no problem with urinary function were obtained with a brief survey preoperatively and postoperatively. RESULTS: At 56 days after removal of urethral catheters, the actuarial rates of urinary continence recovery based on definitions 1 to 3 were 43%, 84% and 82%, respectively. The use of definition 2 for continence resulted in a 1.9 times higher actuarial rate for continence recovery when compared to definition 1 at 56 days (p <0.001). However, strong agreement was observed between definitions 2 and 3 (kappa = 0.69). CONCLUSIONS: Urinary control is recovered in a significant proportion of men who undergo radical prostatectomy during the initial 3 months. Continence rates will vary significantly based on the use of alternative definitions. The clinical practice of asking patients how many pads daily they use may be valid, as it corresponds well to the impairment they have. PMID- 11490255 TI - Quality of life. PMID- 11490256 TI - Inguinal hernia after radical retropubic prostatectomy for prostate cancer: a study of incidence and risk factors in comparison to no operation and lymphadenectomy. AB - PURPOSE: The incidence, mechanisms and risk factors of inguinal hernia after radical retropubic prostatectomy are sparsely elucidated in the literature. We determined the rate of inguinal hernia after radical retropubic prostatectomy and compared it to the incidence in patients with prostate cancer who did not undergo operation or underwent only pelvic lymph node dissection. MATERIALS AND METHODS: We followed 375, 184 and 65 men who underwent radical retropubic prostatectomy plus pelvic lymph node dissection, pelvic lymph node dissection only and no surgery with respect to inguinal hernia for a mean of 39, 47 and 45 months, respectively. The prostatectomy group was also evaluated in regard to the potential risk factors of previous hernia surgery and post-prostatectomy anastomotic stricture. RESULTS: The incidence of hernia was 13.6%, 7.6% and 3.1% in the prostatectomy, lymph node dissection and unoperated group, respectively. The difference was statistically significant in the prostatectomy and unoperated groups according to the Mantel-Cox log rank test and Cox proportional hazards rate. Previous hernial surgery and post-prostatectomy anastomotic stricture were more common in patients with an inguinal hernia after prostatectomy. CONCLUSIONS: The incidence of inguinal hernia is clearly increased in men who have undergone radical retropubic prostatectomy plus pelvic lymph node dissection compared with those who undergo no surgery for prostate cancer. Inguinal hernia appears to develop more often in men with prostate cancer who undergo radical retropubic prostatectomy and pelvic lymph node dissection than in those who undergo pelvic lymph node dissection only. While surgical factors trigger hernial development, previous hernial surgery and post-prostatectomy anastomotic stricture may be important risk factors. In fact, the latter may largely explain the difference in the incidence of inguinal hernia in our lymph node dissection and prostatectomy groups. Prophylactic surgical procedures must be evaluated to address this problem. PMID- 11490257 TI - Risk adjustment for evaluating the outcome of urological operative procedures. AB - PURPOSE: Considerable public and media attention has been directed in recent years toward comparing performance at individual hospitals. So-called death league tables have been published in the media, ranking hospitals according to crude mortality rates. Crude rates of mortality and morbidity are clearly misleading. Therefore, scoring systems comparing treatment outcomes among physicians or hospitals on an objective basis are urgently required. MATERIALS AND METHODS: During a 12-month period we prospectively evaluated 651 patients at 2 urological units using a simple and well validated surgical scoring system. Patients had been admitted to the units for routine urological operations. The scoring system consists of a simple preoperative physiological score, a postoperative severity score and defined types of complications. RESULTS: The morbidity and mortality rates for unit 1 were 7.4% and 1.3%, respectively. For unit 2 the morbidity and mortality rates were 14% and 8.8%, respectively. Despite the marked differences in these crude rates risk adjusted analysis revealed no significant difference (p <0.05). Receiver operating characteristics curve analysis likewise demonstrated no significant difference in mortality and morbidity for the 2 units. CONCLUSIONS: Raw rates of mortality and morbidity are often inappropriately used to compare the performance of various surgical procedures, especially vascular and cardiothoracic surgery. In Great Britain at some institutions urological units are ranked according to mortality and morbidity outcome. As a consequence, important variables, such as patient physiological state at surgery and the type of procedure, are not considered. Our study shows that the scoring system applied is suitable for urological audit and may be a valuable tool for comparing performance at various units. PMID- 11490258 TI - Reconstruction of ureter with transverse retubularized colon segments. AB - PURPOSE: We present a technique in which colon segments are reconfigured as substitutes for ureteral defects. MATERIALS AND METHODS: Via a flank incision a 3 cm. segment was obtained from the ascending or descending colon and reconfigured into a well vascularized tube. In 2 patients a single retubularized colon segment was interposed into an extensive defect of the upper and mid ureter, respectively. In an additional 2 patients 2 combined anastomosed reconfigured colon segments were used for pyelo-colo-cutaneostomy. RESULTS: At a followup of 9 to 27 months (mean 15), no complications related to the reconfigured colon segments were noted. One patient died of progressive disease 9 months postoperatively but with no evidence of ureteral substitute malfunction. CONCLUSIONS: Reconfigured colon segments may be used successfully to reconstruct extensive ureteral defects. Access is via a flank or pararectal incision and intraperitoneal surgery is minimal. The colon segments are obtained immediately proximal to the ureteral defect, necessitating little mobilization of the mesenteric pedicle. Metabolic consequences should be absent or low since only minimal amounts of intestine are isolated. In patients with renal insufficiency or a history of irradiation this technique may be superior to the use of ileum. PMID- 11490259 TI - Glutaraldehyde cross-linked collagen occlusion of the ureteral orifices with percutaneous nephrostomy: a minimally invasive option for treatment refractory hemorrhagic cystitis. PMID- 11490261 TI - Vesical calculus caused by vascular prosthetic graft. PMID- 11490260 TI - Ureteral quadruplication with contralateral triplicate ureter. PMID- 11490262 TI - Iatrogenic pneumocephalus after laparoscopic renal biopsy. PMID- 11490263 TI - Rupture of adrenal carcinoma after biopsy. PMID- 11490264 TI - Local recurrence of an oncocytic adrenocortical carcinoma with ovary metastasis. PMID- 11490265 TI - Complete genitourinary exenteration for multifocal transitional cell carcinoma in renal transplant recipient. PMID- 11490266 TI - Subtotal radical cystectomy after radical prostatectomy: a source of pelvic recurrence of bladder cancer with life threatening bleeding. PMID- 11490267 TI - Squamous cell carcinoma of the phallus in a patient with congenital adrenal hyperplasia. PMID- 11490268 TI - Hemangiopericytoma of the seminal vesicle presenting with hypoglycemia. PMID- 11490269 TI - Undetectable prostate specific antigen in disseminated prostate cancer. PMID- 11490270 TI - Prostatic sarcoma with rapid tumor progression after nerve sparing radical cystoprostatectomy. PMID- 11490271 TI - Local tract metastasis of prostatic adenocarcinoma 8 years after (125)iodine brachytherapy. PMID- 11490272 TI - Arteriovesical fistula: a complication of bladder trauma. PMID- 11490273 TI - Intrarenal varix presenting as an enhancing renal mass with calcifications. PMID- 11490274 TI - Transrectal ultrasound findings of pelvic arteriovenous malformation. PMID- 11490275 TI - Plasma cell granuloma of the testis: unusual localization. PMID- 11490276 TI - Prostatosymphyseal fistula after transurethral resection of the prostate. PMID- 11490277 TI - Tension-free vaginal tape: is it truly tension-free? PMID- 11490278 TI - Reoperation after complicated tension-free vaginal tape procedures. PMID- 11490279 TI - Recurrent bulbar urethral stricture in the region of an artificial urinary sphincter. PMID- 11490280 TI - Female urethral hemangioma. PMID- 11490281 TI - Re: A randomized prospective trial of intrarectal lidocaine for pain control during transrectal prostate biopsy: the Emory University experience. PMID- 11490282 TI - Re: Percutaneous sacral nerve root neuromodulation for intractable interstitial cystitis. PMID- 11490283 TI - Re: Protracted urinary retention necessitating urethrolysis following tension free vaginal tape surgery. PMID- 11490284 TI - Re: The development of erectile dysfunction in men treated for prostate cancer. PMID- 11490285 TI - Re: Is human chorionic gonadotropin useful for identifying and treating nonpalpable testis? PMID- 11490286 TI - Re: Third year medical student attitudes toward learning urology. PMID- 11490287 TI - Histological analysis of urethral healing after tubularized incised plate urethroplasty. AB - PURPOSE: Tubularized incised plate urethroplasty has rapidly gained popularity for treating hypospadias. It is presumed that healing occurs with the postoperative migration of epithelial cells into the incised urethral plate. We describe the time course and histology of the healing urethral wound in an animal model after dorsal incision and stenting. MATERIALS AND METHODS: A procedure was developed for use in an immature porcine model. The ventral aspect of the urethra was opened and a dorsal incision was made in the urethra to the level of the corpus spongiosum. The urethra was then catheterized and closed ventrally. Animals were sacrificed at intervals of 1, 2, 3, 5, 7, 14 and 21 days. Slides were made from multiple cross sections taken from each penis, and stained with hematoxylin and eosin, and Masson trichrome before analysis. RESULTS: Migration of epithelial cells into the dorsal epithelial defect was evident on postoperative day 2 with apparent complete re-epithelialization by postoperative day 5. Regions of increased fibroblastic activity were observed in the subepithelial stroma below the incised area on postoperative day 3 and early collagen deposition was noted in these areas when stained with Masson trichrome. These areas appeared to organize and by postoperative day 21 there was little evidence of increased fibroblastic activity or excess collagen deposition. CONCLUSIONS: Urethral healing after incision and tubularization over a catheter in our model occurred through normal re-epithelialization without excess collagen deposition or scarring. PMID- 11490288 TI - Prepubertal high flow priapism: incidence, diagnosis and treatment. AB - PURPOSE: We reviewed the literature of the last 40 years and report our experience with treating high flow priapism with fistula embolization in prepubertal boys. MATERIALS AND METHODS: Two boys had blunt perineal trauma and 1 had penile trauma (ages 6, 6 and 10 years). Painless priapism developed within 24 hours and lasted for 4 to 7 days before the patients presented to the hospital. Primary diagnosis was made on color Doppler ultrasound. When high flow priapism was diagnosed angiography of the internal iliac artery and embolization of the arteriocavernosal fistula were performed. Mean followup was 26 months. RESULTS: Color Doppler ultrasound revealed bilateral arteriocavernosal fistulas in 2 boys and a unilateral fistula in 1. Angiography showed fistulas of the branches of the internal pudendal artery in 2 patients and fistulas of the bulbourethral artery in 1. Microcoils were used in the bulbourethral artery and a gelatin sponge was used in other penile arteries. Complete detumescence with restored erectile function was achieved in all cases. CONCLUSIONS: High flow priapism in children can be diagnosed easily by typical clinical features combined with color Doppler ultrasound. In children with posttraumatic priapism embolization of the arteriocavernosal fistula is superior to surgical or medical procedures and should be the first line therapy. Embolization using microcoils for bulbourethral arteries and a gelatin sponge for other penile arteries has proved to be safe and successful therapy. PMID- 11490289 TI - Magnetic resonance imaging for the evaluation of hydronephrosis, reflux and renal scarring in children. AB - PURPOSE: We studied the effectiveness of magnetic resonance imaging (MRI) for evaluating hydronephrosis, differential renal function, vesicoureteral reflux and renal scarring in children. MATERIALS AND METHODS: A total of 16 patients with hydronephrosis were evaluated with MRI. Coronal T1 and axial T1 and T2-weighted images were obtained before and after the administration of intravenous contrast material. Patients with vesicoureteral reflux underwent magnetic resonance voiding cystourethrography. Differential renal function was estimated from differential parenchymal volumes determined from MRI using computer software. The results were compared to standard imaging modalities. RESULTS: A total of 19 MRI studies were performed in 16 patients, including 3 for ureteropelvic junction obstruction, 11 for vesicoureteral reflux and 2 for other conditions. MRI provided the best anatomic detail and clear corticomedullary differentiation. MRI identified renal scarring and cortical thinning in 8 cases, while mercaptoacetyltriglycine 3 scans did not show any renal scarring and single photon emission computerized tomography-dimercapto-succinic acid nuclear scans diagnosed only 4 of 5 cases. There was good correlation between differential function obtained from nuclear scans and differential parenchymal volumes obtained from MRI (correlation coefficient 0.86, r2 = 0.74). Magnetic resonance voiding cystourethrography diagnosed reflux in 4 of 5 patients in whom vesicoureteral reflux was previously documented by standard voiding cystourethrography. CONCLUSIONS: MRI provides an alternative for the evaluation of hydronephrosis in children by combining the information provided by functional and anatomic nuclear scans, voiding cystourethrography and ultrasonography in a single study without ionizing radiation. MRI appears to be as good as existing modalities in the evaluation of renal scarring and cortical thinning. PMID- 11490290 TI - Concealed umbilical stoma: long-term evaluation of stomal stenosis. AB - PURPOSE: The technique of forming a concealed umbilical stoma has been described previously and includes a posterior umbilical flap for improved cosmesis and stenosis prevention. We assessed long-term stomal stenosis. MATERIALS AND METHODS: We reviewed retrospectively the charts of 46 patients (mean age at surgery 14 years) of whom 35 had undergone concealed umbilical stoma creation and 11 the Malone antegrade continence enema procedure for continent urinary diversion. Urinary stomas were created from appendix in 20 cases, ileum in 8, sigmoid colon in 5, bladder in 1 and stomach in 1. Malone antegrade continence enema stomas were constructed from appendix in 10 cases and sigmoid colon in 1. A total of 21 patients underwent urinary diversion and augmentation cystoplasty. RESULTS: At followup of 12 to 84 months (median 3.4 years) 93.5% of patients had an intact stoma with no need for surgical revision. Of the remaining patients 3 (6.5%) required revision of the stoma at skin level for stomal stenosis at 1, 4 and 38 months after initial surgery and 2 had a brief period of indwelling catheterization for correction of stenosis. CONCLUSIONS: The concealed umbilical stoma technique provides an excellent cosmetic result with a low rate of stomal stenosis in patients requiring intermittent bladder or bowel catheterization. PMID- 11490291 TI - Outcome of ureteral reimplantation in children with the urge syndrome. AB - PURPOSE: While bladder dysfunction, particularly detrusor-sphincter incoordination, appears to be associated with a higher ureteral reimplantation failure rate, the potential effect of the urge syndrome or detrusor instability alone on surgical outcome remains unclear. We assessed the effect of the underlying urge syndrome on the outcome of ureteral implantation in cases of primary vesicoureteral reflux. MATERIALS AND METHODS: We retrospectively reviewed the charts of all patients who underwent ureteral reimplantation for primary vesicoureteral reflux at our institution in an 11-year period. The urge syndrome was considered frequency, urgency or urge incontinence with absent urinary tract infection. Patients with and without the urge syndrome were compared with respect to the rate of urinary tract infection and renal scarring as well as postoperative outcome. RESULTS: We identified 25 patients (40 ureters) with and 67 (113 ureters) without the urge syndrome. Reimplantation was successful in all except 1 ureter in a patient without the urge syndrome. Postoperatively 2 patients without the syndrome had transient contralateral reflux and 1 in each group had transient ipsilateral reflux. No case required reoperation. Postoperatively patients with the syndrome had a significantly higher incidence of febrile (16% versus 1.5%, p <0.05) and afebrile (52% versus 12%, p <0.05) urinary tract infection. CONCLUSIONS: The results of ureteral reimplantation are excellent with 99.3% success irrespective of the presence or absence of the urge syndrome. Thus, when clinically indicated, reimplantation may be performed safely in such cases. The higher incidence of postoperative urinary tract infection in patients with the urge syndrome may require continuing antibiotic prophylaxis in addition to anticholinergics until voiding symptoms resolve. PMID- 11490292 TI - Outcome analysis of vesicoamniotic shunting in a comprehensive population. AB - PURPOSE: Prenatal detection of obstructive uropathy is used widely and vesicoamniotic shunting is the accepted procedure in well-defined cases. We present outcomes of vesicoamniotic shunting in a consecutive series from a coordinated prenatal-postnatal treatment program. MATERIALS AND METHODS: From 1989 to 1998 bladder obstruction was identified in 89 fetuses at a multidisciplinary prenatal high risk clinic. Prenatal intervention was considered in cases of bilateral hydronephrosis associated with evidence of bladder outlet obstruction and oligohydramnios. RESULTS: Indications for prenatal shunting were present in 12 patients (13%), and 9 underwent vesicoamniotic shunt insertion between weeks 20 and 28 of gestation. None of the prenatal procedures was associated with preterm labor, chorioamnionitis or urine chemistry values greater than cutoff threshold on bladder tap. Shunts were extruded from 2 fetuses, which required sequential insertion. After prenatal intervention 1 patient elected pregnancy termination and the others proceeded to term. Two neonates died at birth, and 6 survived. The underlying urological diagnosis was posterior urethral valves in 4 newborns, urethral atresia in 1, and prune belly variant and urethral atresia in 1. Of the patients 3 had relatively normal renal function, 2 had severe renal insufficiency and 1 had mild renal impairment. Five newborns are voiding freely and 1 underwent bladder reconstruction and is on intermittent catheterization. None of the patients had pulmonary problems at last followup. CONCLUSIONS: Although vesicoamniotic shunting is effective in reversing oligohydramnios, its ability to achieve sustainable good renal function in infancy is variable. No specific prenatal parameters were effective in predicting eventual good renal function. Pulmonary function cannot be assured with restoration of amniotic fluid. PMID- 11490293 TI - Duodenal atresia associated with midgut deletion in cloacal exstrophy: a new association? PMID- 11490294 TI - Extraperitoneal placement of renal allografts in children weighing less than 15 kg. AB - PURPOSE: Extraperitoneal renal transplantation is not routine in small recipients, in whom transperitoneal engraftment is the norm. The outcome of extraperitoneal placement of renal allografts in children weighing less than 15 kg. was evaluated at 2 institutions. MATERIALS AND METHODS: We retrospectively reviewed all pediatric renal transplantations at 2 institutions from 1988 to 2000 and identified 29 children 14 to 72 months old (mean age 29.2) weighing less than 15 kg. (range 8 to 14.8, mean 11.2). All children underwent allograft placement extraperitoneally via a modified Gibson and low midline retroperitoneal incision in 27 and 2, respectively. A concurrent procedure was done via the same incision during 2 ipsilateral and 2 bilateral nephrectomies. RESULTS: Of the 29 patients 25 have a functioning renal allograft. In 2 cases the initial allograft was lost due to early postoperative thrombosis and acute rejection in 1 each. Two patients with a functioning allografts died of medical complications greater than 2 years after transplantation. One child required reexploration secondary to fascial dehiscence and an additional recipient required pyeloureterostomy due to ureteral necrosis after living related donor transplantation. CONCLUSIONS: Extraperitoneal renal transplantation is technically feasible in children who weigh less than 15 kg. This approach preserves the peritoneal cavity, limits potential gastrointestinal complications and allows the confinement of potential surgical complications, such as bleeding and urinary leakage. In addition, this approach provides complete access to the retroperitoneum to enable concurrent retroperitoneal surgery, such as nephrectomy, to be performed safely. We recommend that extraperitoneal renal engraftment should become routine in children weighing less than 15 kg. rather than using the more common transperitoneal approach for allograft placement. PMID- 11490295 TI - Urological complications after renal transplantation using ureteroureteral anastomosis in children. AB - PURPOSE: Ureterovesical reimplantation is most often performed for renal transplantation in children. We reviewed our experience to evaluate the safety and efficacy of ureteroureteral reimplantation in pediatric renal transplantation. MATERIALS AND METHODS: We retrospectively evaluated the charts of 92 boys and 72 girls who underwent a total of 166 ureteroureteral anastomoses for renal transplantation from January 1990 to December 1999. Spatulated end-to end anastomosis was performed between recipient and graft ureters without stenting and with a bladder catheter for at least 10 days. RESULTS: Mean patient age at transplantation was 11.2 years (range 1 to 21.5). There were 22 living related donor and 144 cadaveric grafts. Urological anomalies and nephropathy were the cause of end stage renal disease in 146 and 20 patients, respectively. Urological complications were noted in 14 of the 166 transplantations (8.4%) in 10 boys and 4 girls, including 12 initial and 2 repeat grafts from 2 living related and 12 cadaveric donors. Five of these patients had undergone previous urological surgery. The 2 children (1.2%) with acute ureteral obstruction underwent repeat intervention after stent failure. Anastomotic leakage in 7 cases (4.2%) was treated conservatively in 1 and with a Double-J stent (Medical Engineering Corp., New York, New York) only required in 3. Reoperation was required in 3 cases. One patient (0.6%) with late ureteral stenosis underwent repeat anastomosis, 1 (0.6%) required reimplantation for recurrent pyelonephritis due to vesicoureteral reflux in the graft, 1 (0.6%) with a valve bladder required bladder augmentation and ureteral reimplantation, and 1 (0.6%) with lymphocele and 1 (0.6%) with lithiasis were successfully treated conservatively. Complications were associated with acute rejection in 6 cases. Mean followup without graft loss in patients who presented with versus without complications was 58.3 months (range 1 to 112) versus 75 (range 1 to 118). In the former patients with a mean age of 16 years 9 months versus those without urological complications mean serum creatinine was 116 and 108 mol./l., respectively. Two grafts were lost in patients with urological complications, including 1 who died of pulmonary embolism and 1 with refractory chronic rejection. Seven patients were lost to followup after 54 months (range 12 to 113) of adequate graft function. CONCLUSIONS: Ureteroureteral anastomosis is a safe and effective technique for pediatric renal transplantation with a low complication rate, which may be due to better vascularization of the shorter ureteral end of the graft. Our results should encourage the use of this technique in pediatric renal transplantation. Efforts to preserve the recipient ureters should be made at nephrectomy. PMID- 11490296 TI - Management of grade IV renal injury in children. AB - PURPOSE: Conservative nonsurgical management of major renal trauma in children is well established. However, when blunt trauma is accompanied by significant urinary extravasation, options are less than clearly defined. Endoscopic techniques, such as stents and percutaneous drainage, have not been widely used because of small caliber. We present our experience with endoscopic management of grade IV renal trauma. MATERIALS AND METHODS: From 1983 to 1996, 15 children satisfied the criteria for grade IV renal trauma. We retrospectively reviewed the charts to assess the mechanism of injury, associated injury, treatment, hospital stay and transfusion requirement. Patients were followed clinically with blood pressure and creatinine monitoring, and by radiograph with computerized tomography. RESULTS: Nine patients with isolated kidney injury were successfully treated with observation, 1 underwent early partial nephrectomy for persistent anemia and hypotension, and 5 had a urinoma, which was successfully treated with percutaneous drainage only in 2. The other 3 patients underwent cystoscopy and ureteral stent placement for high drainage output, leading to the resolution of urine leakage. In 1 patient who underwent percutaneous drainage only renovascular hypertension developed, requiring partial nephrectomy 3 months after the original injury. The remaining 13 patients had complete radiographic resolution of the injury and no evidence of hypertension. CONCLUSIONS: In the pediatric population grade IV blunt renal trauma usually resolves without intervention. When a symptomatic urinoma develops, percutaneous drainage, accompanied at times by ureteral stenting provides the complete resolution of persistent urine leakage. PMID- 11490297 TI - Severe hypospadias repair with meatal based paracoronal skin flap: the modified Koyanagi repair. AB - PURPOSE: Various techniques have been reported to repair severe hypospadias. The Koyanagi repair uses a meatal based foreskin flap. We modified this procedure and reviewed the outcome of the repair of severe hypospadias. MATERIALS AND METHODS: The modified Koyanagi repair was performed in 151 children 7 months to 15 years old (mean age 3 years) with severe hypospadias. The records of these patients were reviewed. In all cases the meatus was at or proximal to the penoscrotal junction. Followup was 6 months to 16 years (mean 6 years). Flap design is the same as in the Koyanagi repair, although our modified technique requires removal of the subcutaneous tissue of the distal portion of the flap, which acts as a full-thickness free skin graft. RESULTS: A fistula developed in 19 patients (12.6%), meatal stenosis in 3 (2%) required meatoplasty and infection in 2 (1.3%) resulted in a regressed meatal position. Good cosmetic results were achieved in all except the latter 2 cases. CONCLUSIONS: Our modified Koyanagi repair affords an excellent cosmetic appearance. The complication rate is relatively low and compares favorably with that of other techniques. PMID- 11490298 TI - Sonographic characteristics of the urinary tract in healthy neonates. AB - PURPOSE: In infancy it has been reported that intravesical pressure is generally higher in males than in females. We investigated whether there are significant differences in the sonographic characteristics of the urinary tract in male and female neonates. MATERIALS AND METHODS: The study comprised 280 consecutive full term newborns, including 146 males and 134 females. Kidneys were assessed for hydronephrosis and graded according to Society for Fetal Urology guidelines. Kidney measurements included maximum longitudinal length and largest longitudinal area. Bladder measurements included bladder dimensions and wall thickness. Bladder wall thickness was measured only in neonates with an estimated bladder volume of greater than 10 ml. RESULTS: Of the 280 neonates 114 had mild renal pelvic dilatation in at least 1 kidney and the incidence in males was significantly higher than in females (53% versus 27%). Kidney longitudinal length and largest area were significantly greater in males on each side. Mean bladder volume was essentially equal in the 169 male and female neonates with an estimated bladder volume of greater than 10 ml. but mean bladder wall thickness was significantly greater in males than in females (1.63 versus 1.38 mm.). CONCLUSIONS: Our study shows that there are marked differences in sonographic findings in male and female neonates in regard to renal pelvic dilatation, renal size and bladder wall thickness. These differences should be considered when sonography is done for screening for urinary tract anomalies in newborns. PMID- 11490299 TI - Measurement of urinary flow rate using ultrasound in young boys and infants. AB - PURPOSE: We present a technique for measuring urinary flow rates with ultrasound in male infants and children. MATERIALS AND METHODS: Urinary flow rate was measured simultaneously by an ultrasound probe placed around the base of the penis and by a funnel with a rotating disk at the bottom in 30 boys with a mean age of 6.7 years (range 4.5 to 10.5), and by ultrasound in 8 infants with a mean age of 10 months (range 1 to 28). Voided volume was measured with a graded cylinder or calculated from the weight change of diapers in infants. Ultrasound and rotating disk maximum flow rates were calculated. The ultrasound signal was calibrated by comparing the collected voided volume to the area under the curve for that void. The volume calculated from the rotating disk flow rate curve was also compared with the collected volume. RESULTS: Both methods yielded similar flow curves. However, ultrasound maximum flow rate significantly exceeded rotating disk maximum flow rate (13 +/- 6 ml. per second, range 5 to 22 versus 10 +/- 4 ml. per second, range 4 to 21, t test p <0.001). The underestimation of the flow rate by the rotating disk method may have been due to adherence of urine to the funnel wall. Rotating disk maximum flow rate was lower and voided volume was underestimated by up to 50% (average 15 +/- 2%) in 21 cases. Ultrasound maximum flow rate averaged 6 +/- 3 ml. per second (range 3 to 11.6 [oldest infant]) in the 8 infants. CONCLUSIONS: Urinary flow rates can be measured accurately using ultrasound in boys who produce small volumes and/or who are not toilet trained and also in infants. In future studies ultrasound will be applied to subsets of male infants with bladder dysfunction. PMID- 11490300 TI - Can standard open pediatric urological procedures be performed on an outpatient basis? AB - PURPOSE: We questioned whether it is feasible in the current era of cost consciousness to perform standard open pediatric urological procedures on an outpatient basis while maintaining patient safety and satisfaction. MATERIALS AND METHODS: We report on 51 consecutive patients 2 months to 13 years old (mean age 4 years 3 months) who underwent a standard open pediatric urological procedure between August 1999 and June 2000. The procedures included ureteral reimplantation in 22 cases (tapered in 2), pyeloplasty in 20, partial nephrectomy in 2, nephrectomy in 2, complete ureterocele reconstruction in 1 and other in 4. The expectation that the procedure would be performed on an outpatient basis was discussed with parents preoperatively. We excluded only cases requiring bowel for reconstruction. A caudal block was administered at the start of the procedure using 0.25% bupivacaine with 1:200,000 epinephrine at a dose of 1 cc/kg. The wound was infiltrated with 1 cc/kg. 0.25% bupivacaine and 0.5 mg./kg. ketorolac was administered at the end of the procedure. As soon as the child awakened, an age appropriate diet was started and 0.5 to 1 mg./kg. codeine with acetaminophen was given every 4 hours. RESULTS: Of the 51 children 44 (86%) were discharged home the day of surgery. Average postoperative hospitalization was 7 hours. One of the 44 children discharged home required a single catheterization elsewhere. There were no other complications or repeat hospitalizations. CONCLUSIONS: Our experience shows that standard open pediatric urological procedures may be performed safely and comfortably on an outpatient basis. PMID- 11490301 TI - Novel expression patterns of the myc/max/mad transcription factor network in developing murine prostate gland. AB - PURPOSE: Expression of myc proto-oncogenes and myc-antagonizing mad/mxi genes typically predominate in proliferating versus differentiating cells, respectively. C-myc expression in prostate cells is well established but to our knowledge that of several recently discovered mad/mxi genes is completely uncharacterized. Such characterization is particularly relevant because mxi1 is lost or mutated in some human prostate tumors and mouse mxi1-null mutants show prostatic hyperplasia. MATERIALS AND METHODS: Developing murine prostatic lobes at select postnatal days 1 to 28 were analyzed by in situ immunohistochemical and in vitro RNA analysis. The expression patterns of the 3 myc genes c-, L- and N myc, and the mad1, mxi1 and mad4 genes were studied in most detail with nonradioactive in situ and immunohistochemical analyses. RESULTS: We describe what is to our knowledge previously unreported expression of N- and L-myc in the prostate with particularly the latter strongly expressed throughout development. High c-myc expression was lost at day 7 with re-elevation at day 14, followed by subsequent low expression, representing a unique in vivo confirmation of c-myc expression changes seen previously in several in vitro differentiation systems. The alternatively spliced weak and strong repressor mxi1 isoforms showed distinct, partially overlapping expression patterns. Of particular interest were continual mad1 and mad4 expression during the proliferative and differentiative phases. Similarly mad1 was evident in proliferating normal prostate cell cultures but not in tumor cell lines, suggesting that mad1 expression in prostate may be clinically relevant. CONCLUSIONS: Myc network expression in developing mouse prostate is novel and does not completely fit previous simpler models of Myc versus Mad expression based on other cell types. PMID- 11490302 TI - The influence of sex hormones on renal osteopontin expression and urinary constituents in experimental urolithiasis. AB - PURPOSE: To our knowledge the influence of sex hormones on urinary stone formation remains undetermined. We investigated the effect of castration on urinary lithogenic factors and renal osteopontin expression in rats previously treated with ethylene glycol. MATERIALS AND METHODS: Sprague-Dawley rats were divided normal males, castrated males, males with 2 weeks of 0.75% ethylene glycol treatment, castrated males with 2 weeks of 0.75% ethylene glycol treatment, normal females, castrated females, females with 2 weeks of 0.75% ethylene glycol treatment and castrated females with 2 weeks of 0.75% ethylene glycol treatment. We analyzed 24-hour urine samples for urinary constituents, such as calcium, oxalate, citrate, uric acid, phosphate, magnesium, sodium, potassium and creatinine. The kidneys were examined for osteopontin expression by Northern blot analysis and for crystal deposition by histological examination. RESULTS: In intact male rats calcium and citrate excretion decreased and oxalate excretion increased significantly after ethylene glycol treatment. Castrated male rats with ethylene glycol had greater calcium and less oxalate excretion than male intact rats with ethylene glycol. In intact female rats uric acid excretion decreased and only calcium excretion increased significantly after ethylene glycol treatment. Castrated female rats with ethylene glycol excreted significantly more oxalate and less calcium than intact female rats with ethylene glycol. Renal osteopontin expression was the same in male intact and castrated rats, and in female intact and castrated rats. In males with ethylene glycol expression was stronger in castrated than in intact rats. In females with ethylene glycol expression was weaker in castrated than in intact rats. No crystal deposits were found in the kidneys in any group. CONCLUSIONS: Testosterone appears to promote stone formation by suppressing osteopontin expression in the kidneys and increasing urinary oxalate excretion. Estrogen appears to inhibit stone formation by increasing osteopontin expression in the kidneys and decreasing urinary oxalate excretion. PMID- 11490303 TI - Effects of isoproterenol and butylscopolamine on the friction between an artificial stone and the intraureteral wall in anesthetized rabbits. AB - PURPOSE: We evaluated the effects of the nonselective beta-adrenoceptor agonist isoproterenol and the nonselective muscarinic antagonist butylscopolamine on ureteral wall tension, namely friction between an artificial stone and the intraureteral wall, in anesthetized rabbits. MATERIALS AND METHODS: The relaxing effect of the drugs on the KCl induced tonic contraction was examined in isolated rabbit ureters. The effect of the drugs on the applied force needed for the artificial stone to pass at a constant speed through the ureter, called sliding force, was evaluated in anesthetized rabbits. RESULTS: In a concentration dependent manner isoproterenol but not butylscopolamine reduced the KCl induced contraction in isolated ureter (mean pD2 7.35 +/- 0.06). Intravenous administration of 1 and 10 microg./kg. isoproterenol significantly decreased the friction between the artificial stone and intraureteral wall in anesthetized rabbits with sliding force at 15 minutes after drug administration decreased by 54.5% and 63.6%, respectively. In contrast, 100 and 1,000 microg./kg. butylscopolamine intravenously had no evident effect on ureteral wall tension. CONCLUSIONS: Our results strongly suggest that ureteral smooth muscle relaxation by beta-adrenergic stimulation reduces ureteral wall tension, thereby, diminishing mechanical effects impeding the movement of a ureteral stone down the ureter. PMID- 11490304 TI - Frequently deleted loci on chromosome 9 may harbor several tumor suppressor genes in human renal cell carcinoma. AB - PURPOSE: Loss of various loci on chromosome 9 has been reported in various cancers. To determine the frequency of deletions at different loci of chromosome 9 in renal cell carcinoma microdissected samples of normal renal epithelium and carcinoma from the same patients were analyzed. MATERIALS AND METHODS: DNA was isolated from microdissected sections of normal and tumor cells of 60 renal specimens, amplified by polymerase chain reaction and analyzed for loss of heterozygosity on chromosome 9 using the 16 microsatellite markers D9S178, D9S157, D9S274, D9S168, D9S285, D9S156, D9S1839, D9S162, IFNA, D9S736, D9S171, D9S1749, D9S273D9S270, D9S153 and D9S170. Loss of heterozygosity was analyzed by a polymerase chain reaction based technique developed at our laboratory. RESULTS: This study showed a high incidence of loss of heterozygosity on chromosome 9 in renal cell carcinoma. Of 60 cases 44 (73%), 24 (40%) and 14 (23%) showed loss of heterozygosity at a minimum of 1, at a minimum of 3 and at 4 or more loci, respectively. The main deletion was found on the 9p21 region at loci DS171 in 38% of cases, D9S1749 in 42% and DS270 in 14%. Overall deletion on chromosome 9p21 was noted in 57% of renal cancer cases. Other deleted regions were on chromosome 9p'0022 to 23 at loci D9S157 in 37% of cases, D9S274 in 20%, D9S168 in 27%, D9S285 in 20%, D9S156 in 12%, D9S1839 in 17% and D9S162 in 24%. Overall deletion at chromosome 9q32 to 33 was noted in 46% of renal cell carcinoma cases. Chromosome 9q32 to 33 also showed deletion at locus D9S170 in 22% of renal cell carcinoma cases. When we compared the incidence of deletion at various loci on chromosome 9 according to renal cell carcinoma grade, we found a higher rate of deletion in advanced grades of renal cell carcinoma. A candidate target tumor suppressor gene, p16 (MTS-1/CDKN2), has been identified within the 9p21 deleted region in various cancers. In our study the expression of p16 protein was absent or low in renal cell cancer samples, suggesting that loss of the p16 gene may be involved in renal cell carcinogenesis. CONCLUSIONS: Our study demonstrates a high incidence of loss of heterozygosity on chromosome 9, mainly 9p21 and 9p22 to 23, in renal cell carcinoma, suggesting several putative tumor suppressor genes on these regions. The identification of other tumor suppressor genes on the 9p21 and 9p22 to 23 regions warrants further studies. PMID- 11490305 TI - Potent antitumor effects of CD154 transduced tumor cells in experimental bladder cancer. AB - PURPOSE: Current intravesical immunotherapy for bladder cancer with bacillus Calmette-Guerin instillations is standard treatment for patients with high risk superficial tumors but relapses are common. We evaluated the tumor vaccine concept in murine bladder cancer by comparing tumor cell transduction with genes coding for the immunostimulatory molecules CD154, interleukin (IL)-12 and CD80 to design a novel vaccination strategy. MATERIALS AND METHODS: Adenoviral vectors were used to transduce murine bladder cancer MB-49 cells with genes coding for CD154, IL-12 and CD80. Parental or transduced MB-49 cells were injected subcutaneously into syngeneic mice. The effects of transgene expression on tumorigenicity and the generation of protective immunological memory against challenge with parental tumor were studied. RESULTS: All 76 animals injected with parental MB-49 cells had tumors within 8 to 12 days. Tumor cell expression of CD154 combined with IL-12 completely inhibited tumor outgrowth with all 21 mice tumor-free and CD154 transduction alone was almost as effective with 33 of 35 tumor-free. IL-12 production by tumor cells delayed tumor outgrowth and 4 of 10 mice remained tumor-free. Over expression of CD80 had no effect on tumorigenicity. CD154 expressing tumors were rapidly infiltrated with large numbers of CD4+ and CD8+ T cells. Mice vaccinated 4 times with adenoviral CD154 transduced MB-49 cells were completely protected against challenge with parental tumor. Co-injection of CD154 modified cells with parental MB-49 cells retarded tumor growth. CONCLUSIONS: Our experimental results suggest that the potent antitumor effects of CD154 gene transduction should be considered for immunostimulatory gene therapy for bladder cancer. PMID- 11490306 TI - The effect of antisense Bcl-2 oligonucleotides on Bcl-2 protein expression and apoptosis in human bladder transitional cell carcinoma. AB - PURPOSE: Bcl-2 is an important determinant of transitional cell carcinoma of the bladder recurrence and progression as well as a factor in patient response to chemotherapy or radiotherapy. We determined Bcl-2 down-regulation after antisense oligonucleotide therapy and synergism with mitomycin C in transitional cell carcinoma of the bladder. MATERIALS AND METHODS: Bcl-2 protein was quantified using flow cytometry and immunohistochemistry in 4 bladder cancer cell lines, in bladder washings from 6 patients with carcinoma in situ and in 16 patient tumor samples. The synergistic effects of antisense oligonucleotides G3139 and 2009, and mitomycin C were investigated in 4 cell lines, while 2009 down-regulation was examined in 20 tumor explants in an ex vivo model. RESULTS: Bcl-2 protein expression was found in all 4 cell lines and in 5 of the 6 cell populations derived from patients with carcinoma in situ. Of the 16 tumors 7 were classified positive by frozen section immunohistochemistry and quantitative flow cytometry. G3139 and 2009 down-regulated Bcl-2 protein expression in all 4 cell lines and 2009 down-regulated Bcl-2 protein expression in half of the Bcl-2 positive tumor specimens. There was only evidence in 1 cell line, T24/83, that Bcl-2 protein expression down-regulation enhanced mitomycin C induced apoptotic cell death. CONCLUSIONS: Bcl-2 was expressed in a significant proportion of bladder tumors and in carcinoma in situ. Therefore, antisense oligonucleotides represent a viable strategy for Bcl-2 protein down-regulation. However, it may not always translate into an increased level of mitomycin C induced apoptosis in transitional cell carcinoma of the bladder. PMID- 11490308 TI - Histological and neurotrophic changes triggered by varying models of bladder inflammation. AB - PURPOSE: We determined whether bladder inflammation causes elevated expression of nerve growth factor by bladder parenchymal cells, leading to alterations in neurons innervating the bladder. To answer this question biochemical, histological and neuronal size data were obtained in rats following various experimental models of bladder inflammation. MATERIALS AND METHODS: Chemical (2.5% formalin), immune (lipopolysaccharide 2 x 104 cfu/ml.) and mechanical (chromic catgut) inflammation was evaluated at various times and compared to control bladders. Hematoxylin and eosin, and Giemsa staining was done to characterize inflammation and quantify mast cells in the bladder. Nerve growth factor protein and messenger RNA were assayed in the bladder and major pelvic ganglion using 2-site enzyme-linked immunosorbent assay and reverse transcriptase polymerase chain reaction, respectively. Retrograde axonal tracing was done to size bladder neurons in the major pelvic and dorsal root ganglia. RESULTS: All forms of inflammation increased bladder weight and produced diffuse hyperplasia, intramural edema, acute and chronic inflammatory cells, infiltration and mastocytosis. Generally bladder inflammation resulted in a 50% increase in nerve growth factor and 52% to 58% enlargement of peripheral neurons. CONCLUSIONS: Inflammation results in altered nerve growth factor content of the bladder, and morphological changes in sensory and motor neurons innervating the bladder. Such neuroplasticity may be a possible explanation for the association of bladder inflammation with long-term symptoms and pain after inflammation subsides. PMID- 11490307 TI - Pyridinoline cross-linked carboxyterminal telopeptide of type I collagen as a useful marker for monitoring metastatic bone activity in men with prostate cancer. AB - PURPOSE: We investigated the clinical usefulness of measuring the serum concentrations of pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) and carboxyterminal propeptide of type I procollagen (PICP) as markers for monitoring metastatic bone activity in patients with prostate cancer. MATERIALS AND METHODS: Serum levels of ICTP, PICP, alkaline phosphatase, prostatic acid phosphatase and prostate specific antigen (PSA) were analyzed in 104 untreated patients with prostate cancer, including 62 with and 42 without bone metastasis. Serial measurements of ICTP, PICP and PSA were performed during hormonal therapy in 35 of 62 prostate cancer patients with bone metastasis. RESULTS: Serum levels of all markers except prostatic acid phosphatase were significantly higher with than without bone metastasis. The median values of each marker increased according to the extent of bone metastasis. Serial ICTP, PICP and PSA in 19 patients with a partial response or no change in bone scans demonstrated a downward trend after treatment, while in 16 with progression they showed an upward trend after treatment. The rate of detecting bone metastasis and progression using ICTP were highest compared with other markers based on the percent clinical effectiveness and receiver operating characteristic curves. CONCLUSIONS: Measuring serum ICTP may be useful for detecting bone metastasis and prostate cancer progression, and may augment PSA and bone scan monitoring of metastatic bone activity. PMID- 11490309 TI - Prevention of further cyclophosphamide induced hemorrhagic cystitis by hyperbaric oxygen and mesna in guinea pigs. AB - PURPOSE: Hyperbaric oxygen therapy and mesna have been successfully used for hemorrhagic cystitis. We defined the protective effects of hyperbaric oxygen and mesna in further cyclophosphamide induced hemorrhagic cystitis in guinea pigs. MATERIALS AND METHODS: A total of 48 male guinea pigs were divided into 6 groups. All groups received 2 doses of 68.1 mg./kg. cyclophosphamide intraperitoneally at the same time intervals but group 1 served as controls. Group 2 received cyclophosphamide only, group 3 received hyperbaric oxygen treatment (2.8 ATA for 90 minutes twice daily) before and the day after further cyclophosphamide, group 4 received 21.5 mg./kg. mesna intraperitoneally only with further cyclophosphamide, group 5 received hyperbaric oxygen and mesna with further cyclophosphamide, and group 6 received hyperbaric oxygen before initial cyclophosphamide, between the 2 doses and after the further dose of cyclophosphamide, and mesna on the days of cyclophosphamide. RESULTS: Although mesna alone provided protection against cyclophosphamide induced cystitis in animal bladders, there was also significant damage compared with controls. When the uroprotective efficacy of mesna was supported with hyperbaric oxygen, bladder protection was promoted since mean histological scores and hematuria levels in this group did not differ from those in controls. CONCLUSIONS: According to this animal study using hyperbaric oxygen as adjuvant therapy in humans may be a better tool than mesna alone for the prophylaxis and treatment of cyclophosphamide induced hemorrhagic cystitis. PMID- 11490310 TI - Does cyclooxygenase-2 inhibitor prevent renal tissue damage in unilateral ureteral obstruction? AB - PURPOSE: We determined whether the cyclooxygenase-2 inhibitor etodolac affects renal tubular damage and interstitial fibrosis in unilateral ureteral obstruction. MATERIALS AND METHODS: Etodolac (10 mg./kg.) was administered to rats 1 day before unilateral ureteral obstruction and every day thereafter. Kidneys were harvested at day 14 after unilateral ureteral obstruction. Tissue transforming growth factor-beta and prostaglandin E2 were measured by bioassay using mink lung epithelial cells and enzyme linked immunosorbent-sandwich assay. Renal tubular proliferation and apoptosis were detected by immunostaining with proliferating cellular nuclear antigen and by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling, respectively. Cyclooxygenase-2 expression was detected by immunohistochemistry. Fibrosis was assessed by measuring collagen deposition in trichrome stained slides. RESULTS: Bioassay showed that in the control group obstructed kidneys contained significantly higher mean transforming growth factor-beta1 than unobstructed kidneys (79.1 +/- 8.3 versus 33.6 +/- 4.2 ng./gm. tissue) and etodolac significantly decrease the mean value in obstructed kidneys (46.2 +/- 10.0 ng./gm. tissue). Assay demonstrated that obstructed control kidneys had significantly more mean tubular apoptosis than their unobstructed counterparts (26.6 +/- 5.4 versus 2.2 +/- 1.4 nuclei per high power field) and etodolac significantly decreased mean renal tubular apoptosis in the obstructed kidneys (16.2 +/- 1.9 nuclei per high power field). In addition, immunostaining with proliferating cellular nuclear antigen showed that obstructed kidneys in the control group had significantly more mean renal tubular proliferation than unobstructed kidneys (9.8 +/- 3.4 versus 3.9 +/- 0.1 per high power field) and etodolac significantly increased mean proliferating renal tubule in the obstructed kidneys (24.9 +/- 4.3 per high power field). Control obstructed kidneys had significantly more fibrosis and prostaglandin E2 production, which were also significantly blunted by etodolac. CONCLUSIONS: The cyclooxygenase-2 inhibitor etodolac significantly reduces tissue transforming growth factor-beta, resulting in decreased tubular damage and interstitial fibrosis. This finding suggests that etodolac is a promising agent for preventing renal tissue damage in unilateral ureteral obstruction. PMID- 11490311 TI - Combined evaluation of detrusor pressure and bladder wall thickness as a parameter for the assessment of detrusor function: an experimental in vivo study. AB - PURPOSE: We developed a new method for assessing detrusor function, including active detrusor pressure in relation to bladder wall thickness. This parameter should enable the determination of bladder wall tension, defined as detrusor force per cross-sectional area of bladder tissue. MATERIAL AND METHODS: In acute studies in 8 male foxhounds sacral anterior roots S2 to S3 were placed into a modified Brindley electrode. The bladder was stimulated at different intravesical volumes with bladder filling in 50 ml. stages up to 700 ml. and intravesical pressure was measured and registered. The volume of bladder tissue was determined following cystectomy. Bladder tissue volume and bladder wall thickness were correlated with intravesical pressure during stimulation, resulting in the determination of detrusor force per cross-sectional area in N./cm.2 bladder tissue. RESULTS: The curve of the intravesical pressure rise during stimulation between 0 and 700 ml. showed a rapid incline with a maximum mean of 70.8 cm. water at 100 ml., followed by a slow decline. The bladder wall tension curve had an approximately symmetrical course with a slow incline, a maximum mean of 10.15 N./cm.2 at 350 ml. and a subsequent slow decline. CONCLUSIONS: The different curves of intravesical pressure and bladder wall tension show that these parameters are not identical. Combining detrusor pressure with bladder wall thickness enables further information on detrusor function to be obtained. Therefore, the clinical assessment of bladder wall tension may become a promising supplement to standard diagnostic methods with predictive value in patients with disturbed micturition. PMID- 11490312 TI - Role of ovarian hormones in the pathogenesis of impaired detrusor contractility: evidence in ovariectomized rodents. AB - PURPOSE: Although detrusor hyperactivity with impaired contractility is a common urodynamic finding in elderly subjects, to our knowledge its pathogenesis remains unknown. Biopsy studies indicate that subjects with detrusor hyperactivity and impaired contractility have ultrastructural evidence of dysjunction and degeneration patterns in isolated detrusor hyperactivity and impaired contractility, respectively. Based on the known cellular effects of estrogen we postulated that declines in ovarian hormone production may contribute to the pathogenesis of detrusor hyperactivity with impaired contractility. MATERIALS AND METHODS: Mature 13 to 14-month-old female Fisher 344 rats were studied 4 months after bilateral ovariectomy or sham surgery. Detrusor structure was evaluated by electron microscopy and contractility was evaluated by muscle strip studies. RESULTS: After bilateral ovariectomy detrusor smooth muscle decreased by 25% with a 12% decrease in the number of nucleated muscle profiles and degenerative changes in many axons. Muscle strips from bilaterally ovariectomized animals generated 40% to 50% less tension per strip in response to carbachol than strips of equal size from sham operated animals with no apparent change in muscarinic receptor affinity. CONCLUSIONS: Bilateral ovariectomy resulted in many changes of the degeneration ultrastructural pattern but in none of the characteristic features of the dysjunction pattern. Our results indicate that the mature rodent detrusor and its innervation are sensitive to prolonged ovarian hormonal deficiency, contributing to impaired contractility in rodents. Future studies are required to establish whether estrogen has a role in the degeneration ultrastructural pattern or impaired contractility in humans. PMID- 11490313 TI - Efficacy of the beta3-adrenergic receptor agonist CL-316243 on experimental bladder hyperreflexia and detrusor instability in the rat. AB - PURPOSE: Recent evidence indicates that in a number of species detrusor relaxation is mediated through activation of the beta3-adrenergic receptor. We determined whether activation of the beta3-adrenergic receptor would be a useful therapeutic approach for bladder instability. We profiled in vitro activity of the beta3-adrenergic receptor agonist CL-616243 and the efficacy of this compound in experimental models of detrusor instability and bladder hyperreflexia. MATERIALS AND METHODS: Isolated rat detrusor strips were contracted by depolarizing the preparations with 20 mM. KCl. CL-316423 was added to the tissue bath in increasing concentrations and contraction inhibition was assessed. Efficacy against bladder instability was evaluated using the obstructed hypertrophied bladder model in the rat. The acetic acid bladder cystometry model was used to assess the efficacy of CL-316423 in bladder hyperreflexia. Isovolumetric contractions were evoked by electrical stimulation using a silver bipolar electrode. Data are expressed as the mean plus or minus standard error of mean. RESULTS: CL-316243 inhibited spontaneously contracting, isolated rat detrusor strips in a concentration dependent manner with a mean concentration inhibiting 50% of maximal response of 2.65 +/- 0.36 nM. Intrinsic activity relative to forskolin was 1. In vivo CL-316243 administered intravenously or orally significantly increased the voiding interval and bladder compliance. In addition, there was a decrease in the number of spontaneous contractions during the filling phase in a model of neurogenic and obstruction induced bladder instability. The amplitude of electrically evoked isovolumetric contractions was significantly smaller after CL-316243 exposure. CONCLUSIONS: These data suggest that activating the beta3-adrenergic receptor in rat bladder using CL-316243 may directly inhibit smooth muscle contractility, experimental hyperreflexia and detrusor instability, and be useful for urge urinary incontinence. PMID- 11490314 TI - Expression of neural plasticity related gene in the pontine tegmental area of rats with overactive bladder after cerebral infarction. AB - PURPOSE: We investigated the expression of the neural plasticity related genes c fos, zif268, c-jun, brain-derived neurotrophic factor and tissue plasminogen activator in the pontine tegmental area in rats with overactive bladder induced by cerebral infarction. MATERIALS AND METHODS: Cerebral infarction was induced by left middle cerebral artery occlusion in female Sprague-Dawley rats. Bladder activity was monitored by continuous infusion cystometrography in awake rats. Specimens were obtained from the pontine tegmental area 1, 3, 5, 12 and 24 hours after cerebral infarction or sham operation. The effect of 0.1 mg./kg. intravenously of the N-methyl-d-aspartate glutamatergic receptor antagonist MK 801 on bladder activity, and c-fos and zif268 expression after middle cerebral artery occlusion were studied. Real-time reverse transcriptase-polymerase chain reaction was performed with the LightCycler system (Roche Diagnostics, Mannheim, Germany) to evaluate cerebral infarction influences on gene expression in the pontine tegmental area. RESULTS: Bladder capacity in cerebral infarcted rats was significantly reduced 1 to 24 hours after middle cerebral artery occlusion compared with that of sham operated rats (p <0.05 to 0.01). One hour after occlusion mean c-fos messenger (m)RNA expression plus or minus standard error had significantly increased to 18.9 +/- 4.0 in terms of its density relative to the outer control in a sample obtained immediately after occlusion compared with that in sham operated rats (p <0.05). It returned to the control level within 3 hours after occlusion. Mean zif268 mRNA expression significantly increased to a relative density of 3.2 +/- 1.4 3 hours after middle cerebral artery occlusion (p <0.01) and returned to the control level within 5 hours after occlusion. The expressions of c-jun, brain-derived neurotrophic factor and tissue plasminogen activator was not influenced by occlusion. Pretreatment with MK-801 inhibited bladder overactivity and significantly reduced the expression of c-fos and zif268 mRNA in the pontine tegmental area. CONCLUSIONS: These results indicate that the development of bladder overactivity after middle cerebral artery occlusion is mediated by activation of an N-methyl-d-aspartate receptor and accompanied by an increase in c-fos and zif268 mRNA expression in the pontine tegmental area. PMID- 11490315 TI - The response of fetal sheep bladder tissue to partial outlet obstruction. AB - PURPOSE: We characterized the response of fetal ovine bladder strips to stimulated contraction and relaxation, and compared this response to that of strips from the pregnant mother and those obtained after a short duration of fetal bladder outlet obstruction. MATERIALS AND METHODS: Sham surgery or bladder obstruction was performed in fetal sheep at 90 days of gestation (term 147 days). Bladder tissue was obtained 3 and 5 days later. Isolated strips of full-thickness bladders from fetuses and pregnant females were mounted individually in Tyrode's solution containing glucose. The strips were subjected to electrical field stimulation. Alternate strips were stimulated by adding carbachol, adenosine triphosphate and KCl. Each strip stimulated by carbachol also underwent field stimulation in the presence of carbachol. Relaxation was also tested using isoproterenol and nitroprusside. RESULTS: The response of isolated strips to field stimulation showed phasic contraction or biphasic response, consisting of initial phasic contraction followed by phasic relaxation and a return to control tension after the end of stimulation. In fetal bladder strips field stimulation at all frequencies after carbachol stimulation produced phasic relaxation or a biphasic response with an initial relaxation phase followed by phasic contraction. This field stimulated relaxant response was not present in adult female bladder strips. In addition, field stimulation stimulated relaxation was completely eliminated by pretreatment with N-nitro-L-arginine-methyl ester, indicating that relaxation was nitric oxide mediated. The fetal responses to all forms of stimulation and relaxation were significantly greater than those of pregnant females. After 5 days or greater of obstruction the responses to field stimulation were reduced significantly. In contrast, there were no significant differences in contractile responses to adenosine triphosphate, carbachol or KCl, or the relaxant response to field stimulation after obstruction. However, there was a significant reduction in relaxant responses to isoproterenol and nitroprusside. CONCLUSIONS: In mid gestation sheep fetus contractile responses to field stimulation, adenosine triphosphate, carbachol and isoproterenol are well developed. The fetal ovine bladder shows a strong neuronal nitric oxide response that is not present in the pregnant mother and is maintained after short-term obstruction. PMID- 11490316 TI - Spontaneous Ca2+ activated Cl- currents in isolated urethral smooth muscle cells. AB - PURPOSE: We identified and characterized the membrane currents underlying spontaneous transient depolarization in the urethra. MATERIALS AND METHODS: Myocytes were isolated from sheep urethra by enzymatic digestion and studied by the amphotericin B patch clamp method. RESULTS: Just more than 10% of cells had spontaneous transient inward currents when maintained at -60 mV. Mean amplitude plus or minus standard error of mean of the spontaneous transient inward currents was 102 +/- 35 pA. and mean frequency was 17 +/- 3 minutes-1 in 18 preparations. Within each cell currents sometimes consisted of up to 3 phases but in 16 of 18 cells monophasic spontaneous transient inward currents were also identified. These currents decayed relatively slowly with a mean time constant of 570 +/- 97 ms. Spontaneous transient inward currents were identified as Ca2+ activated Cl- currents because they reversed near the calculated Nernst potential for chloride ions. They were blocked by the Cl- channel blockers 100 microM. niflumic acid and 1 mM. anthracene-9-carboxylic acid as well as in Ca2+-free solution, 10 mM. caffeine and 30 microM. ryanodine. The latter results suggest that spontaneous transient inward currents require intact intracellular Ca2+ stores. Amplitude and frequency were unaffected by 10 microM. nifedipine but were reduced by the nonspecific Ca2+ entry blockers 10 microM. SKF 96365 and 1 mM. La3+. We interpret these results as indicating that the Ca2+ stores underlying the spontaneous transient inward currents may refill by plasmalemmal Ca2+ channels that differ from L-type channels. CONCLUSIONS: Urethral cells fire large spontaneous transient inward currents, mediated by Ca2+ activated Cl- channels, which are adequate to account for the spontaneous transient depolarizations seen in whole urethral tissue. PMID- 11490317 TI - Potassium outward currents in freshly dissociated rabbit corpus cavernosum myocytes. AB - PURPOSE: Cavernous smooth muscle cells have a key role in the control of penile erection and detumescence. In this study the types of smooth muscle cells and currents present in isolated rabbit corpus cavernosum myocytes were characterized. MATERIALS AND METHODS: Immunohistochemical methods were used to identify cavernous smooth muscle cells. Currents were recorded from freshly dissociated myocytes using the whole cell and amphotericin perforated patch clamp techniques. RESULTS: Cavernous myocytes were identified by alpha-smooth muscle actin and smooth muscle myosin immunoreactivity. Based on electrical properties at least 2 types of myocytes were present. Type I cells showed more depolarized membrane potentials, lower capacitance, higher input resistance and increased current densities at positive potentials than type II cells. In types I and II cells at voltages positive to 30 mV, maxi K+ channel (Ca2+ activated large conductance K+ channel or BK) blockade with iberiotoxin or charybdotoxin reduced outward currents by approximately 40% to 80% at 80 mV. Maxi K+ channel blocking did not affect cell membrane potential. Type II cells showed delayed rectifier K+ channel-type outward currents that were not detected in type I cells. Delayed rectifier K+ channel-type currents were resistant to iberiotoxin or charybdotoxin, activated at approximately -50 to -40 mV. and inactivated weakly. CONCLUSIONS: The data suggest that cavernous smooth muscle cells are heterogeneous with at least 2 subtypes identified based on membrane potential, capacitance, input resistance, current density and delayed rectifier K+ channel expression. The activation threshold suggests that delayed rectifier K+ channels are open at the resting membrane potential and, therefore, contribute to control and regulation of the cavernous myocyte excitability. PMID- 11490318 TI - Differences in the antigen pattern recognized by antisperm antibodies in patients with infertility and vasectomy. AB - PURPOSE: Antisperm antibodies may impair sperm fertilizing capacity. They are found in infertile patients and in men after vasectomy. Little is known to date of the biochemical nature of the antigens that induce the production of antisperm antibodies. MATERIALS AND METHODS: Sperm membrane proteins were prepared from donor spermatozoa, separated by 1-dimensional polyacrylamide gel electrophoresis and exposed to seminal plasma samples of 36 infertile men and 34 after vasectomy containing antisperm antibodies. RESULTS: Ten antigenic protein bands with different molecular weight were recognized by antisperm antibodies. Antisperm antibodies binding to the antigen band at 55 kDa. were significantly more common in infertile men, while those binding to the 72 kDa. band were more common after vasectomy. Significant differences also occurred in the incidence of detecting the 55 kDa. antigen band by the antisperm antibodies of patients with and without varicocele. Comparing antisperm antibodies from patients with or without a history of genital diseases or trauma did not reveal significant differences in the antigens detected. CONCLUSIONS: It seems likely that the development of antisperm antibody binding to different antigens is related to the mode of antibody induction. Since the antigenic properties of spermatozoa change during passage through the epididymis, the antigens detected by antisperm antibodies from men with vasectomy are mostly related to epididymal passage. The identification of human sperm antigens is essential for understanding the mechanism by which antisperm antibodies influence the fertilization capacity of spermatozoa. It is also necessary for the potential development of reliable diagnostic methods for antisperm antibodies that are relevant to infertility. PMID- 11490319 TI - Canine model of infertility after spinal cord injury: time course of acute changes in semen quality and spermatogenesis. AB - PURPOSE: We established a canine model of subfertility after spinal cord injury and examined the time course of acute changes in semen quality and spermatogenesis after spinal cord injury. MATERIALS AND METHODS: Seven dogs underwent surgical T7 spinal cord injury. Six dogs were used as controls. Electroejaculation and testicular fine needle aspiration were performed at baseline and twice weekly for 3 weeks after spinal cord injury. Semen quality change was examined by standard semen analysis. Spermatogenesis was assessed by flow cytometry of testicular fine needle aspiration in all dogs as well as by testicular histology at study conclusion in 4 controls and 4 spinal cord injured dogs. RESULTS: No significant changes in spinal cord injured dogs were noted before 3 weeks after injury. From baseline to 3 weeks after injury certain changes were evident in spinal cord injured dogs. Mean antegrade sperm motility decreased from 62.9% to 20.1% (p = 0.008), mean total sperm (antegrade plus retrograde total sperm) decreased from 423 to 294 x 106 which was not statistically significant, and the incidence of testicular haploid cells decreased from 75.6% to 48.3% (p = 0.028). No significant change in any parameter was present in control dogs. The mean number of mature spermatids per cross sectional tubule on final testicular histology was significantly decreased in spinal cord injured dogs compared with controls (13.6 versus 43.9, p = 0.02). CONCLUSIONS: In the canine model tested the dogs readily survived spinal cord injury, electroejaculation was effective for obtaining ejaculate and fine needle aspiration allowed serial examination of spermatogenesis. Three weeks after spinal cord injury but not before 3 weeks sperm motility and spermatogenesis were significantly decreased. However, at the same point this decrease in spermatogenesis was not yet reflected in the total ejaculated sperm count. PMID- 11490321 TI - Insulin glargine (Lantus), a new long-acting insulin. PMID- 11490320 TI - Evidence of an inhibitory effect of diet and exercise on prostate cancer cell growth. AB - PURPOSE: A high fat diet and sedentary lifestyle may predispose men to prostate cancer through effects on serum factors such as hormones. We evaluated the effects of a low fat, high fiber diet and exercise intervention on serum stimulated growth of established prostate cancer cell lines. MATERIALS AND METHODS: Fasting serum was obtained from 13 overweight men before and after undergoing an 11-day low fat, high fiber diet and exercise intervention. Serum was also obtained from 8 men who had complied with the regimen for a mean of 14.2 years. Hormone dependent LNCaP and independent PC-3 prostate cancer cell lines were grown in culture medium containing 10% of subject pre-intervention or post intervention serum and viable cells were counted after 48 hours. Anthropometry, serum free testosterone, lipids and glucose were measured in all subjects. RESULTS: Post-intervention serum from each of the 11-day intervention subjects reduced LNCaP cell growth by a mean of 30% compared with pre-intervention serum from each (p <0.01). LNCaP cell growth in serum from long-term subjects was 15% below that of post-intervention serum (p <0.01). There was no difference in the growth of PC-3 cells when cultured with serum from either intervention group. Serum free testosterone, body weight, glucose and lipids were significantly reduced in 11-day subjects. CONCLUSIONS: A low fat, high fiber diet and exercise intervention resulted in serum changes that significantly reduced the growth of androgen responsive LNCaP prostate cancer cells in vitro. PMID- 11490322 TI - Twinrix: A combination hepatitis A and B vaccine. PMID- 11490323 TI - Color M-mode and pulsed wave tissue Doppler echocardiography: powerful predictors of cardiac events after first myocardial infarction. AB - To assess the association between color M-mode flow propagation velocity and the early diastolic mitral annular velocity (E(m)) obtained with tissue Doppler echocardiography and to assess the prognostic implications of the indexes, echocardiography was performed on days 1 and 5, and 1 and 3 months after a first myocardial infarction in 67 consecutive patients. Flow propagation velocity correlated well with E(m) (r = 0.72, P <.0001). The ratio of peak E-wave velocity (E) to flow propagation velocity also correlated well with E/E(m) (r = 0.87, P <.0001). The positive predictive value of E/FPV > or =1.5 to identify patients with Killip class > or =II was 90%, and the negative predictive value 92%. The corresponding values for E/E(m) > or =10 were 70% and 90%. Cox proportional hazards analysis identified E/flow propagation velocity > or =1.5 (relative risk, 12.4 [95% confidence interval, 4.1-37.3]), E/E(m) > or =10 (relative risk, 11.5 [95% confidence interval, 3.8-34.7]), and Killip class > or =II (relative risk, 7.8 [95% confidence interval, 1.6-40.4]) to be predictors of the composite end point of cardiac death and readmission because of heart failure. Thus flow propagation velocity and E(m) are closely related after myocardial infarction and appear to have similar prognostic information. PMID- 11490324 TI - Detection of early abnormalities of left ventricular function by hemodynamic, echo-tissue Doppler imaging, and mitral Doppler flow techniques in patients with coronary artery disease and normal ejection fraction. AB - We have investigated the possibility of detecting early abnormalities of left ventricular function at the initial phase of ischemic cardiomyopathy. Sixteen normotensive patients with coronary artery disease and normal left ventricular ejection fraction and 6 control patients were studied by invasive hemodynamic techniques in combination with transmitral Doppler flow or with echo-tissue Doppler imaging. The extent of the percentage of left ventricular longitudinal shortening and the systolic peak velocity at echo-tissue Doppler were significantly higher in the control patients than in patients with ischemic cardiomyopathy (P <.01). Left ventricular end-diastolic pressure was higher (P <.05), whereas mean values of isovolumic contraction and relaxation indexes (dP/dt/P: P <.05; +dP/dt: P <.05; -dP/dt: P <.01) were lower in patients with ischemic cardiomyopathy. Tau was significantly longer in ischemic patients (42.7 +/- 8.8 versus 34.5 +/- 3.7 ms, P <.05). In the control patients, the aortic valve closure to peak E interval by transmitral Doppler flow was significantly longer than that measured by echo-tissue Doppler (P <.001), whereas in patients with ischemic cardiomyopathy, this interval difference was still present and significantly shorter (P <.05). In patients with coronary artery disease and normal ejection fraction, minor and early abnormalities of left ventricular function related to isovolumic contraction and relaxation as well as to longitudinal shortening could be detected. In addition, a suction-like effect, detected during early filling evaluation with echo-tissue Doppler, is significantly decreased but not abolished during the early stages of coronary artery disease. PMID- 11490325 TI - Assessment of no-reflow phenomenon after acute myocardial infarction with harmonic angiography and intravenous pump infusion with Levovist: comparison with intracoronary contrast injection. AB - Myocardial contrast echocardiography (intracoronary application) has emerged as an accurate method to detect the "no-reflow phenomenon." To investigate the diagnostic value of harmonic angiography after intravenous infusion of Levovist in assessing "no-reflow," both intracoronary and intravenous contrast injections were performed in a group of patients with acute myocardial infarction. Seventeen consecutive patients with a successfully reperfused acute myocardial infarction within 6 hours of symptom onset were selected for this study. All patients underwent contrast echocardiography with harmonic angiography with Levovist (400 mg/mL, intravenous pump infusion, trigger intervals 1:4 to 1:8) and sonicated albumin (0.5 to 1 mL, intracoronary bolus) on day 1 after the achievement of a sustained coronary reflow. Myocardial perfusion was qualitatively assessed with a 12-segment model. The endocardial length of the residual contrast defect after reflow was also calculated. Forty-four of 204 segments were not analyzed after intravenous contrast echocardiography and 37 after intracoronary contrast echocardiography because of artifacts. Intracoronary and intravenous injections showed a perfusion defect in 31 (19%) segments, with a concordance of 89% (kappa coefficient, 0.72). Concordance in anteroseptal, anterolateral, and inferolateral segments was 95% (kappa = 0.92), 88% (kappa = 0.66), and 83% (kappa = 0.57), respectively. With intracoronary injection used as the reference method, intravenous injection had a sensitivity of 74% and a specificity of 93% for diagnosing contrast defects. The endocardial extent of no-reflow was 18 +/- 19 after intravenous and 21 +/- 17 after intracoronary contrast echocardiography (P = not significant). Intravenous contrast echocardiography with Levovist reliably identifies the no-reflow phenomenon after successful reperfusion, especially in acute anteroseptal myocardial infarction. PMID- 11490326 TI - Transmural variation of myocardial attenuation and its potential effect on contrast-mediated estimates of regional myocardial perfusion. AB - Promising technical developments suggest that it may be feasible to use contrast echocardiography to estimate regional myocardial perfusion. Although the optimal approach has not yet been determined, the use of a nonlinear (harmonic) response of the contrast agent is common to several recent advances. The purpose of this article is to delineate the relation between the anisotropic (angle-dependent) ultrasonic attenuation of the myocardium through which the sound wave has propagated and the regional, nonlinear response of the contrast agent. Apparent perfusion will be modulated by this regionally varying, path-dependent attenuation, which is determined by the local angle between the propagating sound wave and the myofiber orientation. We illustrate the potential magnitude of the effect of myocardial anisotropy for the apical 4-chamber view by examining propagation along the septum and the lateral wall. We present experimentally measured values of the attenuation of excised sheep myocardium, showing statistically significant differences in the attenuation in the mid wall compared with that in symmetrical zones to the left and right of the mid wall, reflecting the well-known myofiber orientations in these 3 regions. The nonlinear (harmonic) response of a contrast agent depends on the local pressure amplitude, which for a given mechanical index is determined by the attenuation accumulated along the path to the point where the regional perfusion is estimated. PMID- 11490327 TI - Radiofrequency spectral analysis of attenuated ultrasound signals in experiments with echo contrast microbubbles. AB - Conventional gray-scale myocardial contrast echocardiography cannot distinguish perfused but attenuated from nonperfused myocardium because both may appear similar at low image intensity. We hypothesized that with radiofrequency spectral analysis of attenuated ultrasound signals, the harmonic-to-fundamental frequency ratio of the peak power spectrum (HFR(P)) could determine the presence of contrast microbubbles. We measured frequency responses of Optison microbubbles at defined degrees of ultrasound signal attenuation with different formulations of silicone (55D, 80A, and 3M); gray-scale intensities of Optison plus water compared with degassed water were analyzed at different attenuation settings ( 25, -32, and -44 dB, respectively). HFR(P) values of Optison plus water were significantly higher than reference values of degassed water at each attenuation setting (55D, -14 +/- 2 dB versus -30 +/- 2 dB, P <.001; 80A, -19 +/- 2 dB versus -30 +/- 3 dB, P <.01; 3M, -22 +/- 2 dB versus -30 +/- 3 dB, P <.05), even though conventional videodensitometric analysis could not distinguish them. HFR(P) analysis objectively detects microbubbles in clinically relevant conditions of attenuation. PMID- 11490328 TI - Stratification of single-vessel coronary stenosis by ischemic threshold at the onset of wall motion abnormality during continuous monitoring of left ventricular function by semisupine exercise echocardiography. AB - We studied the relation between the ischemic threshold at the onset of wall motion abnormality on exercise echocardiography (EE) and the severity of coronary stenosis in patients with 1-vessel coronary artery disease (CAD). We screened 216 consecutive patients who underwent coronary angiography and EE for suspected CAD. Ninety-five (74 men; age, 56 +/- 12 years) satisfied the study criteria, that is, the presence of 1-vessel disease or no evidence of CAD on angiography and a normal baseline echocardiogram. Eighty-seven patients had 1-vessel CAD on angiography, and exercise-induced wall motion abnormality occurred in 73 (77%). Optimal cutoff values of percent diameter stenosis and minimal lumen diameter for predicting a positive EE were 61% (sensitivity and specificity of 76%) and 1.12 mm (sensitivity and specificity of 74%). Among patients with positive EE, heart rate-blood pressure product at ischemic threshold was correlated with quantitative coronary stenosis (r = -0.72, P <.001). The ischemic threshold from continuous monitoring of left ventricular function during semisupine EE is correlated with the severity of coronary stenosis among patients with 1-vessel disease and a normal resting echocardiogram. PMID- 11490329 TI - Four-year prospective study of pulmonary venous thrombosis after lung transplantation. AB - The objective of this study was to prospectively assess pulmonary venous anastomosis by transesophageal echocardiography after lung transplantation. Thrombus formation at the pulmonary venous anastomotic site after lung transplantation may have catastrophic consequences, including allograft failure and stroke. Eighty-seven consecutive adult lung transplant recipients underwent transesophageal echocardiography within 48 hours after surgery. Thrombosis of a pulmonary vein was diagnosed in 13 (15%) of 87 patients in the early postoperative period after lung transplantation. Mean thrombus width was 0.9 +/- 0.4 cm (range, 0.5 to 1.7 cm), with an average peak flow velocity at the site of obstruction of 127 +/- 23 cm/s (range, 90 to 150 cm/s). Five patients with pulmonary vein thrombosis died in the perioperative period, yielding a 90-day mortality rate of 38%. Larger thrombus size and greater acceleration of flow through a narrowed pulmonary vein correlated with poor clinical outcome. During each year of the study, the incidence of pulmonary vein thrombosis declined progressively. Pulmonary vein thrombosis is a potentially ominous complication in the early postoperative period after lung transplantation. Transesophageal echocardiography is a valuable tool for detecting abnormalities of the pulmonary venous anastomosis. Thrombus size and flow velocity at the anastomotic site may guide prognosis and clinical management. Complications of the pulmonary venous anastomosis are in part technical in nature. PMID- 11490330 TI - Left atrial dissection: pathogenesis, clinical course, and transesophageal echocardiographic recognition. AB - Left atrial dissection is an uncommon entity. It is generally associated with mitral valve replacement, but other predisposing factors should be considered in pathogenesis. We discuss a series of 11 patients with pathologically confirmed left atrial dissection who had been diagnosed previously by transesophageal echocardiography. Predisposing factors and surgical or pathologic findings were reviewed to identify the pathogenic mechanism and to explain the clinical course, hemodynamic disorder, and echocardiographic features. Dissection of the coronary sinus secondary to retrograde cardioplegia, endocarditis, cardiac rupture after myocardial infarction, and blunt chest trauma also could be related to its development. Transesophageal echocardiography identified a mobile intimal flap of the atrial wall that was creating a false chamber and allowed accurate diagnosis of prosthetic mitral valve function, endocarditis complications, and a left ventricular pseudoaneurysm after acute myocardial infarction. Color flow Doppler was particularly useful in identifying complications: communication between the false chamber and true left atria, permitting mitral regurgitation through the periannular route; development of atrial shunts; and severe tricuspid regurgitation caused by disruption of the anterior papillary muscle. PMID- 11490332 TI - Caseous calcification of the mitral annulus: a neglected, unrecognized diagnosis. AB - BACKGROUND: Mitral annular calcification is a common echocardiographic finding. Caseous calcification is a rare variant seen as a large mass with echolucencies that resembles a tumor, occasionally resulting in exploratory cardiotomy. The aim of this study was to assess the prevalence of caseous calcification of the mitral annulus, to evaluate patient characteristics and the echocardiographic variables for diagnosing this entity, and to describe the clinical outcome on follow-up of such patients. METHODS: Caseous calcification was defined as a large, round, echo dense mass with smooth borders situated in the periannular region, with no acoustic shadowing artifacts and containing central areas of echolucencies resembling liquefaction. Eighteen patients were diagnosed by 2-dimensional echocardiography as having caseous calcification of the mitral annulus. One had calcification of the tricuspid annulus. Nine patients underwent transesophageal echocardiographic studies. RESULTS: A typical finding of a round, sometimes semilunar, large, echo-dense, soft mass with central echolucencies seen on both transthoracic and in particular transesophageal echocardiography, resembling a periannular mass, was demonstrated. The mass was posteriorly located in all mitral patients. Transesophageal echocardiography added limited information. Three patients underwent mitral valve replacement. The operative findings were a solid mass adherent to the posterior portion of the mitral valve. Sectioning revealed a toothpaste-like, white, caseous material. Sixteen (84%) patients were treated conservatively. On follow-up of 3.8 +/- 2.4 years, 4 patients died of unrelated causes. CONCLUSION: The characteristic appearance of a large, soft, echo-dense mass containing central areas of echolucencies resembling liquefaction at the posterior periannular region of the mitral valve on 2D echocardiography is compatible with the diagnosis of caseous abscess. Such a finding should not be confused with a tumor. Transesophageal echocardiography does not appear to contribute to the diagnosis. This rather impressive lesion appears to carry a benign prognosis. PMID- 11490331 TI - Echocardiography-guided pericardiocentesis with probe-mounted needle: report of 53 cases. AB - Fifty-three pericardiocentesis procedures were performed on 48 patients from 1993 to 2000 at our coronary care unit. Percutaneous puncture (anterior thoracic in 43 cases, subxiphoid in 10 cases) was performed at the site closest to the exploring probe, where the largest amount of fluid was detected. A needle carrier supported by a bracket with two fixed angulations was mounted on the probe. The needle was advanced through the tissues and inside the pericardial space under continuous visualization. The procedure was successful in 52 of 53 cases. In 1 case of diagnostic pericardiocentesis, the pericardial space was impossible to reach because of the minimal amount of pericardial fluid. In 1 case of acute tamponade after transcatheter ablation of the atrioventricular node, the pericardial puncture caused a pleural-pericardial shunt with consequent drainage of pericardial fluid into the pleural space and symptom resolution. In 1 case, a transient atrioventricular type III block occurred. Emergency surgical drainage was not required in any of the cases. No puncture of cardiac walls ever occurred in this series of patients. No major complications occurred; the incidence of minor sequelae was lower than the incidence reported by other studies on pericardiocentesis without continuous visualization. Our technique appears to be safe and easy to perform even in the presence of minimal amounts of pericardial fluid. PMID- 11490333 TI - More than just the "Q-tip sign". AB - In the early days of transesophageal echocardiography, the "Q-tip sign" was occasionally mistaken for a left atrial mass. This area is now well recognized as a normal anatomic landmark dividing the left atrial appendage from the left upper pulmonary vein. However, in this report we describe 2 patients in whom pathology was found in this area, which should not be overlooked. PMID- 11490334 TI - Differential atrial stunning after electrical cardioversion: an echo tissue Doppler case study. AB - Left atrial stunning after cardioversion is a well-known phenomenon. It has been associated with higher risk of postcardioversion thromboemboli and increased risk of recurrence of atrial fibrillation. We present a case of differential atrial stunning after electrical cardioversion for atrial fibrillation. Diagnosis was made by pulsed wave Doppler of mitral, tricuspid, and pulmonary vein inflow and mitral and tricuspid annuli. Differential mechanical atrial stunning may be a common phenomenon after cardioversion and may suggest difference in right and left atrial transport function. Its prevalence needs to be determined by a large study. Doppler tissue imaging might be routinely used in patients after cardioversion for atrial fibrillation to detect atrial stunning. PMID- 11490335 TI - Papillary fibroelastoma of the left atrial appendage. AB - Cardiac papillary fibroelastomas are benign tumors that usually arise from the valvular endocardium. They are clinically important because of their propensity to embolize. We describe, to the best of our knowledge, the first reported case of a papillary fibroelastoma arising from the left atrial appendage, giving rise to multiple cerebral embolic events. The tumor was excised surgically, with no further embolic events. PMID- 11490336 TI - Septal perforator to left ventricular outflow tract fistula. PMID- 11490337 TI - Acquired aorta-pulmonary artery fistula: diagnosis by multiple imaging modalities. AB - Acquired communication between the aorta and the pulmonary artery is a rare phenomenon. We describe two patients with a thoracic aortic aneurysm in whom the diagnosis of a communication with the pulmonary artery was first made on transthoracic echocardiography and then more completely elucidated by means of multiple imaging modalities: transesophageal echocardiography, epiaortic ultrasound, computed tomography, and magnetic resonance imaging. Representative images from these complementary studies are presented. A successful repair of the fistula was subsequently accomplished in both patients. PMID- 11490338 TI - Doppler echocardiographic findings in 2 identical variants of a rare cardiac anomaly, "subtotal" cor triatriatum: a critical review of the literature. AB - Improved diagnostic procedures of echocardiography and better awareness of the anomaly and its possible variants lead the examiner to the right diagnosis of the rare cor triatriatum sinister. We describe two cases of a variant of "subtotal" cor triatriatum sinister. A uniform classification system is desirable to compare cases, treatment, and outcome. PMID- 11490339 TI - Unusual finding of cor triatriatum in a newborn with hypoplastic left heart syndrome. AB - Cor triatriatum is a rare anomaly in which the left atrium is partitioned into 2 distinct chambers with a variable degree of obstruction to left atrial inflow. This anomaly often occurs in isolation but can occur in combination with other cardiac lesions. The association of cor triatriatum with hypoplastic left heart syndrome has not been reported. We describe a newborn with hypoplastic left heart syndrome who also had the unusual finding of cor triatriatum sinistrum. PMID- 11490340 TI - Routine complexity. PMID- 11490341 TI - Role of hypothalamic monoamines in nicotine-induced anorexia in menopausal rats. AB - BACKGROUND: Nicotine reduces body weight by reducing appetite. Estradiol modulates food intake. Menopause or ovariectomy (Ovx) increases food intake and body weight. Nicotine and estradiol individually influence hypothalamic dopamine (DA) and serotonin (5-HT), whose interaction influences food intake and body weight. We investigated whether lower weight gain in menopausal smokers is mediated via changes in hypothalamic DA/5-HT. METHODS: Ovx or sham-operated female rats had 2 microdialysis guide cannulas simultaneously implanted in ipsilateral ventromedial nucleus of hypothalamus (VMN) and contralateral lateral hypothalamic area (LHA). Rats were divided into 4 groups and received a continuous subcutaneous infusion of nicotine or saline Ovx and sham. DA and 5-HT in LHA and VMN were measured by in vivo microdialysis. RESULTS: Nicotine infusion decreased food intake and body weight in Ovx and sham groups. Increase in LHA-DA and VMN-5-HT in sham group occurred with nicotine, whereas an increase in VMN-DA in Ovx groups with and without nicotine and VMN-5-HT in Ovx group with nicotine was observed. CONCLUSIONS: In the presence of estradiol (ovary intact sham operated rats), nicotine lowers food intake and body weight via increased LHA-DA and VMN-5-HT. In menopause (Ovx rats), nicotine lowers food intake and body weight only via increased VMN-DA and 5-HT. Data show that lower weight gain is mediated via changes in hypothalamic monoamines, primarily via ventromedial hypothalamus. PMID- 11490342 TI - NF-kappa B-mediated chemoresistance in breast cancer cells. AB - BACKGROUND: Nuclear factor-kappa B (NF-kappa B) is a known survival pathway, and it may explain differential sensitivity to tumor necrosis factor-alpha (TNF alpha) and chemotherapeutic-induced apoptosis in apoptotically sensitive (APO+) and apoptotically resistant (APO-) Michigan Cancer Foundation-7 breast cancer cells. METHODS: Crystal violet viability and luciferase reporter gene assays were used to determine the inhibitory concentration of viability at 50% (IC(50)) and the inhibitory concentration of activity at 50% (EC(50)) values in APO- and APO+ cells with the selective NF-kappa B inhibitor, BAY 11-7082 (BAY). The apoptotic reporter assay was used to determine the effects of the transfection of the inhibitory kappa B-dominant negative (I kappa B-DN) construct in conjunction with TNF, paclitaxel, or doxorubicin treatments in these cells. RESULTS: The concentrations at which 50% of cell viability is inhibited (IC(50)) and at which 50% of NF-kappa B activity is inhibited (EC(50)) for BAY in APO- and APO+ cells were 95.24 micromol/L and 1.53 micromol/L, respectively, and 7.62 micromol/L and 2.64 micromol/L, respectively. The IC(50) and the EC(50) values were equivalent for the APO+ cells (P =.665), but not for the APO- cells (P =.025). I kappa B-DN- transfection alone, or with TNF, doxorubicin, or paclitaxel treatments resulted in cell death of both APO- and APO+ cells as compared with vector-control; however, greater cytotoxicity was seen in the APO+ cells. Direct comparison of the APO+ cells versus the APO- cells revealed that these differences were significant (P =.05). CONCLUSIONS: Pharmacologic or molecular inhibition of the NF-kappa B pathway blocked cell survival in MCF-7 APO+ cells, while only molecular inhibition induced cytotoxicity in the APO- cells. Selective manipulation of the NF-kappa B pathway in combination with standard chemotherapeutic agents may lead to an increased potency and efficacy of these agents. PMID- 11490343 TI - Factors that predict the presence of sentinel lymph node metastasis in patients with melanoma. AB - BACKGROUND: This analysis was performed to identify prognostic factors that are predictive of sentinel lymph node (SLN) metastasis in melanoma. METHODS: Analysis was performed of a multi-institutional, prospective, randomized trial of SLN biopsy for melanoma. Eligibility criteria included age 18 to 70 years, Breslow thickness of 1.0 mm or more, and clinically negative regional lymph nodes. SLNs were evaluated by serial sectioning and immunohistochemistry for S100. Univariate chi-square and multivariate logistic regression analyses were performed to assess factors predictive of the presence of a positive SLN. Probability values of less than.05 were considered significant. RESULTS: SLNs were identified in 99.7% of patients. A total of 1058 patients were evaluated; 961 patients had complete data and were included in the statistical analysis. SLNs were positive for tumor in 208 of 961 patients (22%). Breslow thickness, Clark level, ulceration, and patient age were factors that were found to be independently predictive of the presence of SLN metastasis. CONCLUSIONS: Increasing Breslow thickness, Clark level of more than III, the presence of ulceration, and patient age of 60 years or less are the most important independent prognostic factors associated with the finding of positive SLN in patients with melanoma. PMID- 11490344 TI - Synergistic induction of IL-10 by hypertonic saline solution and lipopolysaccharides in murine peritoneal macrophages. AB - BACKGROUND: Liver injury after ischemia/reperfusion is an important cause of morbidity in surgical patients. We have shown that the preconditioning of animals that were subjected to liver ischemia/reperfusion with hypertonic saline solution (HTS) prevented injury by inhibiting Kupffer cell tumor necrosis factor (TNF) production. We postulated that the induction of anti-inflammatory interleukin-10 (IL-10) by HTS might contribute to protection. METHODS: Murine thioglycolate- elicited peritoneal exudative macrophages (PEMs) were used to model the effects of HTS on IL-10 release from Kupffer cells. Cells were preconditioned with 500 mOsm HTS (or isotonic saline medium) for 2 hours and then stimulated with lipopolysaccharide (LPS; 1 microg/mL) or vehicle for 4 hours under isotonic conditions. TNF-alpha and IL-10 were measured in the culture supernatant by enzyme-linked immunosorbent assay; TNF, IL-10, and SOCS-3 messenger RNA expression were assessed by Northern blot. NF-kappa B activation was examined by electrophoretic mobility shift assay and Western blot for I kappa B degradation. RESULTS: In the absence of LPS, isotonic medium--and HTS-pretreated PEMs produced little IL-10 (24.9 +/- 66.0 and 0 pg/mL, respectively); however, stimulation of PEMs with LPS increased IL-10 (134.9 +/- 72.2 pg/mL). Preconditioning with HTS significantly augmented LPS-induced IL-10 production, resulting in a 2-fold increase in IL-10 compared with the isotonic solution LPS group (270.7 +/- 106.8 pg/mL; P <.01). HTS alone increased IL-10 mRNA levels and markedly augmented levels induced by LPS alone. To determine whether IL-10 accounted for HTS-induced TNF inhibition, cells from IL-10 knockout animals were studied. A lack of IL-10 did not reverse the inhibitory effect of HTS on LPS-induced TNF. NF-kappa B activation was the same in HTS-and isotonic solution--pretreated groups after LPS. CONCLUSIONS: HTS augments IL-10 induction by LPS at the gene level. Although TNF is reduced, it is not causally related to increased IL-10 or altered NF-kappa B signaling. HTS might exert its beneficial effects by independently modulating pro- and anti-inflammatory molecules, accounting for the potent immunomodulation exerted by HTS in vivo. PMID- 11490345 TI - Gene transfer of the Bcl-2 gene confers cytoprotection to isolated adult porcine pancreatic islets exposed to xenoreactive antibodies and complement. AB - BACKGROUND: Exposing adult porcine pancreatic islets (PI) to xenoreactive natural antibodies (XNA) induces brisk inflammatory injury that involves activation of the complement system. Gene transfer of Bcl-2 has been shown to protect PI from apoptosis and necrosis in several models. In this study, we investigated the effect of Bcl-2 gene transfer on protection of PI from primate XNA and complement mediated injury. METHODS: The PI were isolated from adult female sows. Only islet preparations that exhibited >90% viability and purity were used. Fresh rhesus monkey serum served as the XNA source. Gene transfer of Bcl-2 was achieved with an adenoviral vector (AdBcl-2) at 500 particle forming units (pfu)/cell. The Bcl 2 expression was confirmed by Western blot technique. Untransfected and transfected PI were incubated in 50% fresh complete serum (CS) or heat inactivated (HI) rhesus serum for 24 hours. The PI viability was analyzed with acridine orange and ethidium bromide staining. Antibody and complement-mediated cytotoxicity were tested by intracellular lactate dehydrogenase (LDH) release. The PI function was assessed in vitro by static incubation studies and in vivo after intraportal transplantation in diabetic severe combined immunodeficiency (SCID) mice. RESULTS: The AdBcl-2 gene transfer resulted in Bcl-2 gene expression in >90% of PI cells. Following exposure to XNA, <15% of the untransfected cells were viable. Similar results were obtained in PI transfected with a similar recombinant adenovirus encoding the reporter gene E coli beta-galactosidase (AdLacZ), an irrelevant gene. A significant increase in LDH release was observed in control PI after exposure to CS compared with PI that overexpressed Bcl-2 (82.89% +/- 7.78% vs 34.31% +/- 5.4%, P <.005). Higher insulin release was observed in vitro in PI transfected with Bcl-2 compared with untransfected PI or islets transfected with AdLacZ (stimulation index of 0.9 +/- 0.31, 0.9 +/- 0.3 vs 2.67 +/- 0.4, respectively). Only PI treated with AdBcl-2 were able to achieve euglycemia after exposure to XNA and complement after transplantation. CONCLUSIONS: Transfer of the antiapoptotic and antinecrotic Bcl-2 gene into PI can reduce primate XNA and complement-mediated lysis. Cytoprotection of PI with Bcl-2 has potential to improve survival of PI xenotransplants. PMID- 11490346 TI - Regional effects of nafamostat, a novel potent protease and complement inhibitor, on severe necrotizing pancreatitis. AB - BACKGROUND: We evaluated the effect of the novel protease inhibitor nafamostat on rat necrotizing pancreatitis through different routes of administration. METHODS: Three hours after the induction of severe pancreatitis, the rats received intravenous gabexate or intravenous or local mesenteric intra-arterial nafamostat. At 9 hours, ascites and bronchoalveolar lavage fluid were collected for the evaluation of capillary leakage (Evans blue extravasation). Pancreas and lung were excised for histologic features, myeloperoxidase, and trypsinogen activation peptide. Twenty-four hour survival was evaluated. RESULTS: Only the intravenous infusion of nafamostat significantly reduced myeloperoxidase (11.7 +/ 2.3 vs 18.3 +/- 1.8 mU/mg; P <.05) and capillary leakage in lungs (Evans blue dye, 1.6 +/- 0.3 vs 2.6 +/- 0.3; P <.05). Only intra-arterial infusion of nafamostat significantly diminished capillary peritoneal leakage (Evans blue dye, 3.6 +/- 0.9 vs 9.4 +/- 0.4; P <.01). Typsinogen activation peptide levels were significantly reduced in all groups, but only intra-arterial infusion did so to baseline. Histologic inflammation in the pancreas was most significantly reduced after intra-arterial infusion (0.92 +/- 0.08 vs 2.91 +/- 0.06; P <.05). No form of protease inhibition reduced mortality rates. CONCLUSIONS: The effects of protease inhibition depend on the route of administration. Nafamostat has maximal effects on the pancreas and peritoneal capillary leakage when delivered by way of local intra-arterial infusion, and shows a greater reduction of lung leukocyte infiltration and capillary leakage by the intravenous route. Nafamostat is more effective than gabexate. PMID- 11490347 TI - Oscillation in the activities of MEK/ERK1/2 during cardiopulmonary bypass in pigs. AB - BACKGROUND: Because cardiopulmonary bypass (CPB) is associated with edema and vasoreactive dysfunction and ERK1/2 pathway is involved in vascular contractility and permeability, a time course study was performed to monitor MEK/ERK1/2/Elk-1 activities during CPB. METHODS: Pigs were subjected to normothermic CPB for 90 minutes followed by post-CPB perfusion for 180 minutes. Atrial myocardium was sampled before CPB, 5 minutes after CPB onset, 5 minutes after weaning from CPB, and at the end of post-CPB. Skeletal muscle and mesenteric vessels samples were harvested before CPB, 5 minutes after CPB institution, and every 30 minutes thereafter to the end of post-CPB. Samples were analyzed by immunoblotting and immunofluorescence microscopy with the use of specific antibodies against active (phosphorylated) forms of ERK1/2, MEK1/2, and Elk-1. RESULTS: Pigs that were subjected to CPB showed an increase in phospho-ERK1/2 after 30 minutes of CPB, followed by a decrease after 90 minutes. Another phosphorylation peak was observed 30 to 60 minutes of post-CPB, followed by a decrease to below baseline at the end of reperfusion. MEK1/2 and Elk-1 activation profiles paralleled ERK1/2 activity peaks. Control samples showed no significant increase above basal levels. CONCLUSIONS: Activation of MEK/ERK1/2/Elk-1 pathways closely follows major CPB surgical manipulations (institution and termination) and could be related to morbidity during and after CPB. PMID- 11490348 TI - Accelerated internalization and detoxification of endotoxin by anti lipopolysaccharide antibody is an Fc receptor--mediated process. AB - BACKGROUND: Interaction between lipopolysaccharide (LPS), LPS-binding protein, and the CD14 receptor at the surface of LPS-responsive cells results in inflammatory cytokine release and internalization and detoxification of LPS. Monoclonal antibodies (mAbs) raised against the deep-core lipid A or the O-linked polysaccharide moieties of LPS accelerate internalization and detoxification of LPS without stimulating cytokine release. This study was conducted to test the hypothesis that the antibody-mediated internalization of LPS is an Fc receptor (FcR)--mediated process. METHODS: Fluoroisothiocyanate (FITC)-conjugated Escherichia coli O111:B4 LPS was incubated with RAW 264.7 cells and allowed to internalize for 2 hours in the presence and absence of anti-LPS, anti-CD14, and isotype control mAbs, and Fab fragments from the anti-CD14, anti--Fc receptor, and control mAbs. Tumor necrosis factor--alpha (TNF-alpha) release was measured by WEHI 164 cell bioassay. FITC-LPS uptake was measured by flow cytometry. Statistical analysis was by analysis of variance and Fisher exact test. RESULTS: Addition of anti-LPS antibodies resulted in a 30- to 40-fold acceleration of LPS internalization (P <.01) in agreement with previous studies. This increase was blunted by anti-CD14 and also by isotype control holo-antibody (P <.01), but not by Fab fragments from anti-CD14 or isotype control antibody. Both anti-FcR antibodies and Fab fragments blocked anti-LPS antibody--stimulated uptake of FITC LPS. Both intact anti-CD14 holo-antibody and Fab fragments blocked TNF-alpha release (P <.01). CONCLUSIONS: Clearance and detoxification of LPS are thought to be essential to the host response to endotoxin. It has been shown that antibodies to LPS accelerate its internalization by monocytic cell lines without increasing the elaboration of cytokines. We found that specific blockade of CD14 by Fab fragments could block TNF-alpha release but not alter the accelerated internalization of LPS produced by anti-LPS antibodies. In contrast, a nonspecific blockade of internalization was produced by competing antibody, which suggests a mechanistic role for the FcR. Specific blockade of FcR by either holo antibody or Fab fragments blocked accelerated internalization, which confirms a FcR mechanism. We conclude that the accelerated internalization of LPS produced by anti-LPS antibody is an Fc receptor--mediated process. These results have significance for the development of adjuvant immunotherapy for gram-negative bacterial sepsis. PMID- 11490349 TI - Phospholipase A(2)--derived neutral lipids from posthemorrhagic shock mesenteric lymph prime the neutrophil oxidative burst. AB - BACKGROUND: Our previous work identified posthemorrhagic shock mesenteric lymph (PHSML) lipids as key elements in polymorphonuclear neutrophil (PMN)--provoked acute lung injury. We hypothesize that gut phospholipase A(2) (PLA(2)) is responsible for the generation of proinflammatory lipids in PHSML that primes circulating PMNs for enhanced oxidative burst. METHODS: Mesenteric lymph was collected from rats (n = 5) before (preshock), during the induction of hemorrhagic shock (mean arterial pressure, 40 mm Hg x 30 minutes), and at resuscitation (shed blood + 2x lactated Ringer's solution). PLA(2) inhibition (quinacrine, 10 mg/kg, intravenously) was given before shock was induced. Extracted lipids were separated by normal phase high-pressure liquid chromatography and resuspended in albumin. PMNs were exposed to a 5% vol:vol concentration of eluted lipids and activated with N-formyl-methionyl-leucyl phenylalanine (1 micromol/L). Superoxide production was assessed by cytochrome C reduction. RESULTS: High-pressure liquid chromatography--extracted neutral lipids of lymph collected before hemorrhagic shock did not prime the PMN oxidase, whereas isolated neutral lipids of postshock lymph primed PMNs 2.6- +/- 0.32-fold above baseline (P <.05). PLA(2) inhibition returned PHSML neutral lipid priming to baseline levels. CONCLUSIONS: PLA(2) inhibition before hemorrhagic shock abrogates the neutrophil priming effects of PHSML through reduction of the accumulation of proinflammatory neutral lipids. Identification of these PLA(2) dependent lipids provides a mechanistic link that may have therapeutic implications for postshock acute lung injury. PMID- 11490350 TI - Apolipoprotein J inhibits the migration and adhesion of endothelial cells. AB - BACKGROUND: Apolipoprotein J (ApoJ) is expressed after vascular injury and remodeling and may inhibit endothelial cell activation in the vascular wall. Recently, ApoJ was identified as upregulated in hyperplastic lesions after prosthetic arterial grafting. This study analyzed the effect of ApoJ on human umbilical vein endothelial cell (HUVEC) migration, adhesion, and proliferation. METHODS: Cell migration towards ApoJ + fetal bovine serum (FBS) or vascular endothelial growth factor (VEGF) was evaluated with the use of a microchemotaxis chamber with or without a fibronectin-coated membrane. For migration that involved fibronectin, cells were exposed to ApoJ before or after placement on the membrane. Cell adhesion to fibronectin was studied similarly but without stimulant. The vital dye alamar blue assessed proliferation of ApoJ + FBS- or VEGF-stimulated HUVECs. RESULTS: ApoJ alone did not cause migration or proliferation of HUVECs. Without fibronectin, ApoJ decreased the migration of HUVECs towards FBS or VEGF. When fibronectin was introduced, ApoJ decreased cell migration toward FBS or VEGF and decreased adhesion only when HUVECs in solution were exposed to ApoJ before the placement on fibronectin. ApoJ had no effect on FBS- or VEGF-induced proliferation. CONCLUSIONS: ApoJ inhibits HUVEC migration and adhesion. By altering endothelial function during vascular injury, ApoJ appears to regulate, in part, the early development of intimal hyperplasia after prosthetic arterial grafting. PMID- 11490351 TI - CD8(+) T cells express a T-helper 1--like phenotype after burn injury. AB - BACKGROUND: Previous studies suggest that CD8(+) T cells are immunosuppressive after burn injury, but recent reports indicate that CD8(+) T cells have several functions similar to CD4(+) T cells, including the secretion of cytokines. This study uses HY male antigen in transgenic HY female mice to determine the antigen specific response of activated CD8(+) T cells after burn injury. METHODS: HY TCR transgenic female mice underwent burn or sham injury. Seventy-two hours after the burn, splenocytes were stimulated with 20 micromol/L HY peptide for 16, 48, and 64 hours; cellular proliferation, intracellular interferon-gamma and interleukin 2, and apoptosis were measured. RESULTS: Burn injury significantly impaired proliferation to HY antigen (P < or =.05). Activated CD8(+) T cells from burned mice showed increased intracellular interferon-gamma and interleukin-2 16 hours after stimulation compared with sham (P < or =.05) and at no time was less than control mice. The percent of CD8(+) T cells decreased with the time of stimulation but was not due to apoptosis by Annexin V staining. CONCLUSIONS: Activated CD8(+) T cells express a T(h1)-like phenotype after burn injury. This provides evidence that CD8(+) T cells are not simply suppressive and that is consistent with data that CD4(+) T cells are primed for a T(h1) response after burn injury. PMID- 11490352 TI - Immunogenicity of L(d+) transgenic mouse hearts. AB - BACKGROUND: C57BL/6 mice transfected with the L(d) gene coupled to the alpha myosin heavy chain promoter result in transgenic mice with L(d) antigen expressed only on cardiac tissue. These transgenic animals allow the examination of immune reactivity against cardiac L(d) by "self" or by adoptively transferred L(d) specific 2C cells, and the response of nontransgenic C57BL/6 mice to the transplanted L(d+) heart. METHODS: Naive cardiac L(d+) transgenic mice were examined for evidence of L(d) "autoimmunity." Forty million fresh 2C cells or 2C cells sensitized in vitro for 7 days against Balb/c (L(d+)) + interleukin-2 were also given intravenously to L(d+) transgenic mice. At 5 and 12 days after injection, heart-infiltrating lymphocytes were analyzed by fluorescence-activated cell sorter. The L(d+) transgenic hearts were also transplanted to syngeneic L(d ) nontransgenic C57BL/6 to evaluate the heart's immunogenicity. RESULTS: Naive L(d+) transgenic mice did not exhibit any evidence of lymphocytic infiltration on histologic examination. Adoptive transfer of either fresh or in vitro sensitized 2C cells was also unable to reject the native L(d+) heart in transgenic mice (100% of the mice survived long term [more than 60 days]). Sensitization of the L(d+) transgenic mice with a Balb/c skin graft and interleukin-2 pump infusion (7 days) beginning 1 day before 2C cell injection also did not promote rejection of the native L(d+) heart. However, fluorescence-activated cell sorter analysis did reveal that a significantly greater number of in vitro sensitized 2C cells homed to the L(d+), but not L(d-), heart after both 5 and 12 days (P <.01, P <.001). In contrast, C57BL/6 mice rejected the L(d+) (C57BL/6 background) transgenic heart in a mean survival time of 17 +/- 9.7 days (P <.01), whereas a syngeneic C57BL/6 heart transplant was accepted indefinitely. Lymphocytic infiltration consistent with rejection was present in all animals receiving an Ld+ transgenic heart transplant, whereas no infiltrate was present in those receiving a syngeneic C57BL/6 heart transplant. CONCLUSIONS: Although the class I L(d) transgene is not recognized in its native host, its immunogenicity is shown by the homing of anti L(d) 2C cells to the heart in situ and rejection of L(d+) heart grafts when transplanted into syngeneic C57BL/6 mice. PMID- 11490353 TI - The role of iNOS in wound healing. AB - BACKGROUND: We have previously shown that the blockade of nitric oxide (NO) synthesis impairs wound healing, in particular collagen synthesis. Conversely, impaired wound healing is accompanied by decreased wound NO synthesis. Fibroblast collagen synthesis, proliferation, and fibroblast-mediated matrix contraction are critical to wound healing. We examined the wound healing-related phenotypic changes that are induced by the loss of inducible nitric oxide synthase (iNOS) gene function in fibroblasts. METHODS: Dermal fibroblasts were obtained from 8- to 12-week-old iNOS--knock out (KO; C57BL/Ai-[KO] Nos2 N5) and wild type mice by an explant technique and used after 1 to 3 passages. Proliferation ([(3)H] thymidine incorporation) and collagen synthesis ([(3)H]-proline incorporation into collagenase-sensitive protein) were studied after stimulation with 10% fetal bovine serum. Matrix remodeling was assessed by the measurement of the contraction of fibroblast-populated collagen lattices. RESULTS: iNOS-KO fibroblasts proliferated more slowly, synthesized less collagen, and contracted fibroblast-populated collagen lattices more slowly than wild-type fibroblast. Collagen synthesis was restored to normal in KO fibroblasts in response to NO donors (s-nitroso-N-acetylpenicillamine). CONCLUSIONS: iNOS deficiency causes significant impairment in wound healing-related properties of fibroblasts, which suggests that NO plays an important role in wound healing. PMID- 11490354 TI - Adenosine analogue reduces spinal cord reperfusion injury in a time-dependent fashion. AB - BACKGROUND: We hypothesized that inflammation during spinal cord reperfusion worsens ischemic injury. ATL-146e, an adenosine A(2A) agonist with known anti inflammatory properties, was used to test this hypothesis at varied intervals to determine the time course of reperfusion injury. METHODS: Forty rabbits underwent cross-clamping of the infrarenal aorta for 45 minutes. One group (n = 14 animals) received 0.06 microg/kg/min systemic ATL-146e over 3 hours, beginning after 30 minutes of ischemic time. A second group (n = 6 animals) received ATL-146e over 1.5 hours. A third group (n = 3 animals) received ATL-146e over 1 hour, and a fourth group (n = 17 animals) received saline solution. All animals were assessed at 48 hours for hind limb motor function (Tarlov scale, 0-5). RESULTS: Animals that received ATL-146e for 3 hours (Tarlov score, 4.3 +/- 0.22; P <.001) or 1.5 hours (Tarlov score, 2.7 +/- 0.6; P <.05) had improved neurologic outcomes compared with rabbits that received saline solution (Tarlov score, 0.6 +/- 0.29). Animals that received ATL-146e for 1 hour (Tarlov score, 0.7 +/- 0.8) were not significantly different from those animals that received saline solution. CONCLUSIONS: Systemic ATL-146e, given during reperfusion, results in time dependent improvement in spinal cord function after ischemia. This implies that the mechanism of spinal reperfusion injury includes leukocyte-mediated inflammation at a critical post-ischemic time interval. PMID- 11490355 TI - Both serotonin and a nitric-oxide donor cause chloride secretion in rat colonocytes by stimulating cGMP. AB - BACKGROUND: Previous studies have demonstrated that an antagonist of nitric oxide synthase inhibits neurally mediated chloride secretion in response to serotonin (5-HT). The purpose of this study was to demonstrate that chloride secretion in rat colonocytes that were caused by stimulation of neural 5-HT receptors is mediated by way of a nitrergic pathway that involves the activation of guanylate cyclase. METHODS: The nitric oxide (NO) donor, diethylenetriamine/NO (DNO), was added to an enriched suspension of rat colonocytes that were preloaded with (36)Cl(-). In parallel experiments, DNO (1 micromol/L) was added to cells that were pretreated with the specific inhibitor of soluble guanylate cyclase, NS2028 (2 micromol/L). In additional studies, the neural 5-HT(3) receptor agonist, 2 methyl-5-HT (10 micromol/L), was added to the serosal surface of muscle-stripped sheets of rat colonic mucosa that were mounted in Ussing chambers under voltage clamp conditions, both in the absence and presence of NS2028 (20 mircro). RESULTS: DNO induced 18.0% +/- 8.0% greater (36)Cl(-) efflux than controls (P <.05; n = 14 animals). This efflux was abolished by previous treatment with NS2028. In the chamber experiments, 2-methyl-5-HT induced electrogenic chloride secretion that was significantly inhibited by previous treatment with NS2028 (2.2 +/- 0.5 microA/cm(2) vs 13.1 +/- 2.1 microA/cm(2); P <.001; n = 9 animals). CONCLUSIONS: The predominant secretomotor neurotransmitter that mediates the chloride secretory effects of 5-HT in vitro is nitric oxide. Both the secretory effect initiated at the 5-HT(3) receptor on enteric neurons and at the NO(-) receptor on the rat colonocytes are mediated through the activation of intracellular guanylate cyclase and the production of cyclic guanosine monophosphate. PMID- 11490356 TI - Inhibition of p38 mitogen activated protein kinase increases lipopolysaccharide induced inhibition of apoptosis in neutrophils by activating extracellular signal regulated kinase. AB - BACKGROUND: Prolonged polymorphonuclear neutrophil (PMN) survival has been implicated in tissue injury after sepsis. Previously we reported that lipopolysaccharide (LPS) inhibits PMN apoptosis via the activation of the extracellular signal-regulated kinase (ERK). Conversely, the p38 mitogen activated protein kinase (MAPK) pathway is involved in the spontaneous apoptosis of PMNs. The interaction between these 2 pathways and their ability to regulate apoptosis during sepsis remain largely undefined. We hypothesize that there is interaction between the ERK and p38 pathways during sepsis. METHODS: PMNs were isolated from healthy volunteers by Ficoll gradient centrifugation and red blood cell sedimentation. Cells were then pretreated for 1 hour with the ERK inhibitor (PD98059, 10 micromol/L), p38 inhibitor (SB203580, 1 micromol/L), or vehicle. After treatment with LPS, apoptosis and MAPK activity were correlated. RESULTS: LPS stimulation significantly inhibits PMN apoptosis compared with unstimulated cells. Furthermore, inhibition of ERK significantly abrogates this effect, whereas inhibition of p38 augments LPS induced inhibition of apoptosis. Elk-1 phosphorylation (ERK target) is significantly increased by LPS alone and by inhibition of the p38 pathway during LPS stimulation. This correlates with ERK phosphorylation by Western blot. CONCLUSIONS: These data show that p38 inhibition enhances ERK activity during endotoxemia. Furthermore, these data suggest that cooperation between ERK and p38 MAPK pathways dictates the apoptotic potential of PMNs during inflammatory states. PMID- 11490357 TI - Prostaglandin E2 receptors EP2 and EP4 are down-regulated in human mononuclear cells after injury. AB - BACKGROUND: Recent characterization of prostaglandin receptor subtypes shows that each is critical to cellular functions and operates through separate signaling pathways that may explain differing effects of prostanoids. This study aimed to determine whether prostaglandin receptors EP2 and EP4 are modulated after injury and to evaluate the effect of prostaglandin E(2) (PGE(2)) addition and blockade on EP receptor expression. METHODS: Peripheral blood mononuclear cells (PBMCs) isolated from 10 patients sustaining fracture or burn injury and 10 control subjects were stimulated with lipopolysaccharide +/- NS-398, an inhibitor of PGE(2) production. Samples were evaluated for production of PGE(2), tumor necrosis factor--alpha, and leukotriene B(4) as well as mRNA expression of EP receptors and COX-2. EP receptor expression was also evaluated after treating control PBMCs with PGE(2). RESULTS: PBMCs from injured patients exhibited significant increases in PGE(2) production and COX-2 mRNA compared with control subjects, and these increases were inhibited by NS-398. In contrast, EP2 and EP4 receptors were markedly down-regulated after injury and NS-398 restored expression to control levels. Decreased EP2 and EP4 receptor expression after injury was replicated by coincubation of PBMCs with PGE(2). CONCLUSIONS: Specific PGE(2) receptors are down-regulated after injury and NS-398 reverses this response. Furthermore, PGE(2) mediates EP2 and EP4 down-regulation. These data suggest that specific EP receptor subtypes may provide critical targets for augmenting the immune response after injury in humans. PMID- 11490358 TI - Catecholamines decrease nitric oxide production by cytokine-stimulated hepatocytes. AB - BACKGROUND: Catecholamines are significantly elevated in inflammatory responses and play a regulatory role in sepsis. Nitric oxide (NO), also a key inflammatory mediator in sepsis, is produced in large amounts by the inducible nitric oxide synthase (iNOS) in the liver. The purpose of this study was to test the hypothesis that catecholamines play a role in the regulation of NO production by hepatocytes. METHODS: Primary hepatocytes were isolated from healthy male Sprague Dawley rats and either cultured with normal medium or stimulated with cytomix (interleukin-1 beta, interferon-gamma, and tumor necrosis factor-alpha) in the presence or absence of epinephrine or norepinephrine at varying concentrations. Total RNA was isolated 6 hours after treatment and analyzed by Northern blotting for iNOS mRNA. Protein extracts were obtained at 12 hours and were analyzed by Western immunoblotting for iNOS. Cell culture supernatants were analyzed for NO, determined as the stable end-product NO(2)(-), at 24 hours. RESULTS: Epinephrine and norepinephrine significantly decreased NO(2)(-) levels in stimulated hepatocytes but had no effect on iNOS mRNA or protein levels. The decrease in NO(2)(-) was reproduced by the adenylate cyclase stimulator, forskolin. The catecholamine-induced decrease in NO(2)(-) was completely reversed by the protein kinase A inhibitor Rp-8-Br-cyclic adenosine monophosphate. CONCLUSIONS: Catecholamines decrease hepatocyte production of NO in response to cytokine stimulation. This effect seems to be due to post-translational events and appears to be mediated in part by cyclic adenosine monophosphate. PMID- 11490359 TI - Butyrate sensitizes human colon cancer cells to TRAIL-mediated apoptosis. AB - BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a novel member of the tumor necrosis factor family, induces apoptosis in TRAIL sensitive tumors through the activation of the caspase pathway. Sodium butyrate (NaBT) induces differentiation and apoptosis in certain colorectal cancers; the molecular mechanisms for these effects have not been clearly defined. The purpose of our study was to determine whether NaBT sensitizes TRAIL-resistant human colon cancer cells to the effects of TRAIL. METHODS: Human colon cancer cells (KM12C, KML4A, and KM20) that are resistant to TRAIL treatment alone were treated with TRAIL (100 ng/mL), NaBT (5 mmol/L), or a combination of these agents and harvested for total RNA and protein. Western blots were performed to assess intracellular expression of Flice-like inhibitory protein (FLIP), a caspase inhibitor. Percent-specific apoptosis, relative caspase-3 activity, and Annexin-V immunofluorescence were determined at 24 and 48 hours. Cell cycle--related gene expression was assessed by RNase protection. RESULTS: Treatment with NaBT for 24 and 48 hours decreased FLIP protein expression in all cell lines. Furthermore, NaBT sensitized these resistant cancer cells to the effects of TRAIL with significant increases noted in cell death, caspase-3 activity, and Annexin-V staining compared with NaBT alone. CONCLUSIONS: Our findings suggest that the reduction of FLIP protein levels by NaBT renders TRAIL-resistant human colon cancer cells sensitive to TRAIL-mediated apoptosis. The combination of TRAIL with agents (such as NaBT, which target proteins that prevent cell death) may provide a more effective and less toxic regimen for the treatment of resistant colon cancers. PMID- 11490360 TI - Influence of trauma system implementation on process of care delivered to seriously injured patients in rural trauma centers. AB - BACKGROUND: Statewide trauma systems are implemented by health care policy makers whose intent is to improve the process of care delivered to seriously injured patients. In Oregon, Advanced Trauma Life Support (ATLS) training was mandated for all physicians employed in the emergency department of trauma centers. The purpose of this study was to test the hypothesis that mandatory ATLS training favorably influenced processes of care. METHODS: Seriously injured patients treated at 9 rural Level 3 and Level 4 hospitals were studied before (PRE) and after (POST) implementation of Oregon's trauma system. The processes of care evaluated on the basis of chart review were 20 diagnostic and therapeutic interventions advocated in the ATLS course. A cumulative process score (CPS) between 0 and 1 was assigned on the basis of the processes of care delivered. A CPS of 1 indicated optimal process of care. RESULTS: Mean CPS for 506 PRE period patients (0.44 +/- 0.27) was significantly lower than the mean CPS for 512 POST period patients (0.57 +/- 0.27) with an unpaired t test (P <.001). For the subgroup with injury severity score of 16 to 34, the mean CPS of survivors (0.67 +/- 0.19) was significantly higher than the mean CPS of decedents (0.57 +/- 0.25). CONCLUSIONS: Process of care for seriously injured patients improved after categorization of rural trauma centers in Oregon. Evidence shows improved process of care may have benefitted patients with serious but survivable injuries. Measurement of process of care is an alternative to mortality analysis as an indication of the quality of care. PMID- 11490361 TI - CaSm/gemcitabine chemo-gene therapy leads to prolonged survival in a murine model of pancreatic cancer. AB - BACKGROUND: CaSm, the cancer-associated Sm-like oncogene, is overexpressed in greater than 80% of pancreatic tumors. We previously reported that an adenovirus expressing antisense RNA to CaSm (Ad-alpha CaSm) can decrease pancreatic tumor growth in vivo but is not curative. In the current study we investigated the mechanism of Ad-alpha CaSm's antitumor effect to rationally approach combinatorial therapy for improved efficacy. METHODS: AsPC-1 and Panc-1 human pancreatic cancer cells were treated with Ad-alpha CaSm and examined by MTT assay for in vitro proliferation changes. Flow cytometry determined the effect of CaSm down-regulation on the cell cycle, and then cells treated with Ad-alpha CaSm in combination with cisplatin, etoposide, or gemcitabine chemotherapies were reexamined by MTT assay. SCID-Bg mice bearing subcutaneous AsPC-1 tumors were treated with Ad-alpha CaSm, gemcitabine, or the combination and monitored for tumor growth and survival. RESULTS: Treatment with Ad-alpha CaSm reduced the proliferation of AsPC-1 and Panc-1 cells (59% and 44%, respectively; P <.05). The cell cycle revealed a cytostatic block with decreased G(1) phase and increased DNA content in treated cells. The combination of Ad-alpha CaSm with gemcitabine significantly reduced in vitro proliferation (66% vs 39% and 48% for controls), decreased in vivo AsPC-1 tumor growth by 71% (n = 10), and extended survival time from 57 to 100 days. CONCLUSIONS: Down-regulation of CaSm reduces the growth of pancreatic cancer cells by altering the cell cycle in a cytostatic manner. The combination of Ad-alpha CaSm with gemcitabine is more effective than either agent used separately. PMID- 11490362 TI - Measuring service-specific performance and educational value within a general surgery residency: the power of a prospective, anonymous, Web-based rotation evaluation system in the optimization of resident satisfaction. AB - BACKGROUND: We used a Web-based evaluation system to institute specific changes to various clinical teaching services in our integrated residency in an effort to optimize the overall quality of the educational experience and measured the resident satisfaction in these rotations. METHODS: Residents rated 8 categories of experience on a scale of 1 to 5 (maximum summation score, 40 points). Data were analyzed by t-test for equality of means. A probability value of less than.05 was considered significant. RESULTS: Compliance with completion of the evaluations was 100%. The Chronbach's alpha reliability coefficient of the tool was 0.826. Tukey's estimate of power to achieve additivity was 1.5. Six under performing services were re-engineered with prominent effects on 7 postgraduate year (PGY) rotations. On 2 general surgery services at 1 hospital, the workload was redistributed, and a dedicated team teaching time was instituted (PGY-3 [a]: before, 22 points/after, 31 points; P =.003; PGY-3 [b]: before, 25 points/after, 31 points; P =.004; PGY-1: before, 24 points/after, 29 points; P =.07). A general surgery service at another hospital redistributed coverage of the attending surgeons to create a nonteaching service (PGY-1: before, 22 points/-after, 27 points; P =.01). The transplantation service (PGY-3) was examined, and the role of the point was redefined (before, 24 points/after, 31 points; P =.01). One vascular service (PGY-2) redistributed cases and workload (before, 27 points/after, 22 points; P =.07). The vascular PGY-2 position was eliminated and replaced by a mid-level practitioner. The cardiothoracic service (PGY-1) rotation was converted into a preceptorship (before, 23 points/after, 30 points; P =.015). CONCLUSIONS: A web-based clinical rotation evaluation provides a means for the assessment of the impact of programmatic changes while preserving resident anonymity and maintaining accountability. PMID- 11490363 TI - Targeting platelet aggregation: CD39 gene transfer augments nucleoside triphosphate diphosphohydrolase activity in injured rabbit arteries. AB - BACKGROUND: CD39, the major endothelial nucleoside triphosphate diphosphohydrolase (NTPDase), plays an important role in local thromboregulation. We hypothesized that balloon injury (BI) leads to an acute reduction in arterial NTPDase activity that could be restored by a targeted gene delivery strategy. METHODS: Recombinant adenoviral vectors containing human CD39 (Ad-CD39) or beta galactosidase (Ad-LacZ) were used. Endothelial (ECs) and smooth muscle cells (SMCs) were infected in vitro and NTPDase activity measured. New Zealand white rabbits (N = 28) underwent bilateral iliofemoral artery balloon injury, followed by incubation with Ad-CD39, Ad-LacZ, or vehicle. Explanted vessels were analyzed for NTPDase activity and localization of CD39 expression by immunohistochemistry. Deposition of fluorescent-labeled platelets was studied 3 days after injury and vector treatment. RESULTS: In vitro, Ad-CD39 infection resulted in a greater than 40-fold increase in adenosine diphosphatase activity in ECs and a 3-fold increase in SMCs. In vivo, CD39 transgene expression localized to the luminal aspect of Ad CD39--treated vessels. BI resulted in an acute reduction in vessel wall NTPDase activity (P <.05). Ad-CD39 augmented NTPDase activity when compared with vehicle or Ad-LacZ (P <.05). Platelet deposition on the injured arterial surface was modest and not different between Ad-CD39-- and Ad-LacZ--treated vessels. CONCLUSIONS: BI decreases native NTPDase activity, which can be augmented by adenovirus-mediated gene transfer of CD39. Further studies are required to determine whether targeted delivery of CD39 could convey thromboprotective properties to an injured vessel. PMID- 11490364 TI - Morphine directs T cells toward T(H2) differentiation. AB - BACKGROUND: Failure of cell-mediated immunity is thought to increase the morbidity and mortality rates after trauma and major surgical procedures and to be the result, in part, of a redirection of CD4(+) T cells toward T(H2) differentiation. We tested the hypothesis that morphine treatment after injury promotes T(H2) differentiation of precursor T cells through the mu-opioid receptor. METHODS: Human peripheral blood mononuclear cells (PBMCs) or splenocytes from either wild type or mu-opioid receptor knock-out mice were treated in vitro with either vehicle or morphine and then stimulated with anti CD3/anti-CD28. The supernatant was assayed for T(H1) (interleukin-2 [IL-2], interferon gamma [IFN gamma]) and T(H2) (IL-4, IL-5) cytokines (enzyme-linked immunosorbent assay). Morphine regulation of IL-4 transcription was investigated in PBMCs (IL-4 messenger RNA, nuclear factor of activated T-cells) and Jurkat T cells transfected with a murine IL-4 promoter-luciferase construct. Morphine induced nuclear factor of activated T-cell (NFAT) binding was assayed with the electromobility shift assay in Jurkat T cells. RESULTS: Morphine treatment of PBMCs decreases IL-2 and IFN gamma and increases IL-4 and IL-5 as a function of morphine concentration. Morphine treatment in wild type splenocytes inhibited IFN gamma and stimulated IL-4 protein synthesis. Changes in cytokine synthesis were abolished in mu-opioid receptor knockout mice. Morphine treatment increases IL-4 messenger RNA accumulation in PBMCs and increases IL-4 promoter activity in Jurkat T cells. Morphine increases NFAT nuclear protein binding to an NFAT DNA response element. CONCLUSIONS: We conclude that morphine treatment promotes T(H2) differentiation through a mu-opioid receptor mechanism and that morphine treatment increases IL-4 transcription, in part, through an NFAT mechanism. PMID- 11490365 TI - Protein kinase epsilon dampens the secretory response of model intestinal epithelia during ischemia. AB - BACKGROUND: Luminal fluid sequestration and diarrhea are early manifestations of mesenteric ischemia. This can be modeled in vitro with the use of T84 intestinal epithelia, where ischemia induces Cl(-) secretion with adenosine-mediated autocrine feedback. Protein kinase C (PKC) regulates epithelial transport and, in some organ systems, is involved in the response to ischemic stress. The purpose of this study was to define the role of PKC on epithelial transport during ischemia. METHODS: By voltage-current clamp, short-circuit current (Isc) equals Cl(-) secretion. Ischemic conditions were simulated with the use of a well established chemical hypoxia protocol. RESULTS: Chemical hypoxia briskly activated Isc. Go6850, an antagonist of novel and conventional PKC isoforms, markedly enhanced the ischemia-induced Isc response, although Go6976 (which inhibits only conventional isoforms) had no effect. Rottlerin, a specific inhibitor of PKC delta, did not attenuate ischemic Isc. Both phorbol 12 myristate, 13-acetate and bryostatin-1, which selectively activate PKC epsilon in T84 cells, markedly attenuated the Isc response to ischemia. Both agents also inhibited the Isc response to exogenous adenosine. CONCLUSIONS: PKC (likely the novel epsilon isoform) in intestinal epithelia modulates ischemia-induced alterations in ion transport. Inhibition of PKC epsilon exaggerates the secretory response that is induced by ischemia and by authentic adenosine; conversely, augmented activation of PKC epsilon inhibits secretion. Manipulation of PKC epsilon could limit luminal fluid sequestration during mesenteric ischemia. PMID- 11490366 TI - Vascular cell adhesion molecule--1 expression is obligatory for endotoxin-induced myocardial neutrophil accumulation and contractile dysfunction. AB - BACKGROUND: Sepsis-induced cardiac dysfunction occurs commonly in critically ill patients and is associated with high mortality rates. Neutrophils play a central role in sepsis-induced lung and liver injury; however, the mechanism of sepsis induced cardiac dysfunction remains unclear. Vascular cell adhesion molecule-1 (VCAM-1) has been implicated in neutrophil-mediated liver injury during endotoxemia and is also expressed in myocardium. The purposes of this study were to examine the temporal relationship of myocardial VCAM-1 expression with neutrophil accumulation during endotoxemia and to determine whether VCAM-1 mediates neutrophil accumulation and cardiac dysfunction during endotoxemia. METHODS: Mice were subjected to lipopolysaccharide (LPS; 0.5 mg/kg, intraperitoneally). Myocardial VCAM-1 expression and neutrophil accumulation were determined by immunofluorescence staining. Cardiac performance with or without VCAM-1 blocking antibody (5 mg/kg, intravenously) was determined by the Langendorff technique. RESULTS: LPS caused a time-dependent increase in both myocardial VCAM-1 expression and neutrophil accumulation. At 6 hours after LPS, the immunofluorescent intensity for VCAM-1 increased from 2.5 +/- 0.6 x 10(6) in saline solution controls to 19.9 +/- 3.5 x 10(6) (P <.05, analysis of variance), and neutrophil count increased from 2.4 +/- 1.7/mm(2) in saline solution controls to 13.0 +/- 2.5/mm(2) (P <.05). Left ventricular developed pressure was decreased maximally at 6 hours after LPS compared with saline solution controls (29.1 +/- 1.1 mm Hg vs 53.1 +/- 3.9 mm Hg; P <.05). Treatment with VCAM-1 monoclonal antibody abrogated both myocardial neutrophil accumulation and cardiac dysfunction during endotoxemia. CONCLUSIONS: LPS-induced myocardial dysfunction is associated with increased expression of VCAM-1 and with neutrophil accumulation. Blockade of VCAM-1 abrogates myocardial neutrophil accumulation and preserves cardiac function during endotoxemia, which supports a role for VCAM-1 as a therapeutic target for myocardial protection during sepsis. PMID- 11490367 TI - Nitric oxide mediates dendritic cell apoptosis by downregulating inhibitors of apoptosis proteins and upregulating effector caspase activity. AB - BACKGROUND: Dendritic cells (DCs) play a crucial role in the amplification of the immune response by promoting antigen presentation, T-lymphocyte proliferation, and proinflammatory cytokine and nitric oxide (NO) production. We have previously shown that the exogenous NO donor, s-nitroso-N-acetyl-penicillamine, promotes DC apoptosis by disrupting the mitochondrial membrane potential, which induces cytochrome-C release and activates caspase 3. To further elucidate the signaling pathway, we examined the expression of cellular inhibitors of apoptosis proteins (cIAPs) and poly (ADP-ribose) polymerase cleavage (PARP), a terminal event in the apoptotic cascade. METHODS: DC2.4 were exposed to 250 micromol/L s-nitroso-N acetyl-penicillamine for various intervals. Apoptosis and necrosis were measured by terminal deoxynucleotidyl transferase nick-end labeling assay or flow cytometry with Annexin V and propidium iodide. DC2.4 were cultured with the pan caspase inhibitor, ZVAD (100 micromol/L). cIAP, pro-caspases, and PARP expression or activation was measured by Western blot. Caspase enzyme activity was confirmed with the use of specific substrates. RESULTS: NO-induced DC apoptosis correlated with the downregulation of cIAP expression. Caspase 3 and 6 were upregulated by SNAP and significantly inhibited by ZVAD. Maximal PARP cleavage occurred at 8 hours and coincided with the downregulation of cIAP and peak caspase 3 and near maximal caspase 6 activity. CONCLUSIONS: NO-induced DC apoptosis is associated with the downregulation of cIAP expression, which facilitates caspase cascade activation and subsequent PARP cleavage. PMID- 11490368 TI - A tripartite anoikis-like mechanism causes early isolated islet apoptosis. AB - BACKGROUND: This study examines the mechanisms of early isolated islet apoptosis (II-APO) and loss of functional islet mass. METHODS: Rhesus islets were isolated for transplantation, and an aliquot was used for in vitro molecular studies of II APO. These studies used Western blotting to examine caspase activation and perinuclear envelope protein cleavage that are associated with II-APO and used immunofluorescence analysis of Annexin V and mitochondrial permeability index to examine spontaneous and tripartite anoikis-like (TRAIL) mechanism--induced II APO. RESULTS: Caspase 6 was prominently activated in association with spontaneous II-APO, which occurred after overnight culture. In contrast, caspase 7, 8, and 9 were not activated. Cleavage of focal adhesion kinase and Lamin, substrates of caspase 6, was also evident in spontaneous II-APO. II-APO was exaggerated by the addition of the TRAIL mechanism. The TRAIL mechanism--induced II-APO was blocked by the caspase 6 inhibitor, VEID, and by the soluble fusion proteins, DR4 or DR5, which act as decoy receptors. In vivo studies in diabetic severe combined immunodeficiency disease mice showed that rhesus islets were cytoprotected by either ex vivo gene transfer of Bcl-2 or treatment of the isolated islet with VEID. CONCLUSIONS: These studies suggest 3 major mechanisms involved in II-APO: caspase 6 activation, a TRAIL-induced apoptosis pathway, and the mitochondrial associated apoptosis pathway. Inhibition of these II-APO pathways may improve isolated islet survival and reduce functional islet mass loss, which compromises the stable reversal of diabetes. PMID- 11490369 TI - Inhibition of Fas signaling prevents hepatic injury and improves organ blood flow during sepsis. AB - BACKGROUND: Fas/Fas ligand (FasL) system is one of the major pathways triggering apoptosis that has been shown to play an important role in development and pathogenesis of various diseases including liver and gastrointestinal diseases. Studies indicate that FasL deficiency provides a survival advantage in mice subjected to polymicrobial sepsis. However, the extent to which Fas/FasL contributes to organ injury during sepsis is unclear. Thus, the aim of this study was to determine whether in vivo administration of a Fas-signaling inhibitor during sepsis preserves organ function. METHODS: Male adult C3H/HeN mice were subjected to cecal ligation and puncture (CLP) or sham CLP (sham). Twelve hours after CLP, mice received either Fas-receptor fusion protein (FasFP) (200 microg/kg body weight) or the saline vehicle. Twenty-four hours after the onset of sepsis, cardiac output and organ blood flow were measured with radioactive microspheres. Plasma levels of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase were assessed as indexes of liver damage. Changes in systemic cytokines were measured by enzyme-linked immunosorbent assay. RESULTS. The data indicate that although cardiac output and organ blood flow in the liver, intestine, kidneys, spleen, and heart decreased markedly at 24 hours after CLP, treatment with FasFP maintained the measured hemodynamic parameters and improved hepatic, intestinal, and heart blood flow (P <.05) and partially restored spleen and renal blood flow. Moreover, FasFP treatment markedly attenuated the systemic rise in alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and interleukin 10 (P <.05). CONCLUSIONS: These results not only indicate that there is a role for Fas/FasL mediated processes in the induction of organ injury but suggest that inhibition of Fas/FasL pathway may represent a novel therapeutic modality for maintaining organ perfusion and preventing liver injury during sepsis. PMID- 11490370 TI - The "July phenomenon" and the care of the severely injured patient: fact or fiction? AB - BACKGROUND: The "July phenomenon," a common belief in medical academia, refers to purported errors, inefficiency, and negative outcomes during the summertime transition of the house staff. We hypothesized that care in a trauma service is consistent throughout the year and that the July phenomenon therefore is a myth. METHODS: The records of adults admitted to a trauma service between July 1994 and September 1999 were evaluated. The care of and outcomes for patients admitted in July and August were compared with those of patients admitted in April and May. RESULTS: Nine hundred seventeen patients were evaluated over 5 years. Patients were well matched by the Injury Severity Score, the Glasgow Coma Score, by mechanism, and by survival probability. Patients admitted in the spring were significantly older, by a mean of 5.1 years. Length of stay and intensive care unit stay were similar. Emergency department times were similar, as were resuscitation times, infection rates, and hospital costs. The mortality of patients was similar between the 2 times. CONCLUSIONS: There was no evidence of an increase in negative outcomes early in the academic year compared with the end of the academic year. We believe that a systematic approach to the diagnosis, resuscitation, and treatment of trauma prevented a July phenomenon. PMID- 11490371 TI - Results of a phase I trial evaluating a liver support device utilizing albumin dialysis. AB - BACKGROUND: We have previously reported the clearance of protein-bound and water soluble hepatic toxins, in vitro and in an animal model, using albumin dialysis as an extracorporeal hepatic support (ECHS) device. OBJECTIVE: The objective of this study was to evaluate albumin dialysis through a phase I clinical trial. We hypothesized that albumin dialysis would (1) decrease elevated levels of hepatic toxins, (2) increase the Fischer ratio, and (3) decrease hepatic encephalopathy (HES) and intracranial pressure (ICP), while (4) maintaining stable hemodynamics. METHODS: Patients with acute liver failure were treated with an ECHS device utilizing continuous hemodiafiltration with continuous albumin dialysis. Mean arterial blood pressure (MAP), heart rate (HR), systemic venous oxygen saturation (Svo(2)), ICP, and HES were recorded. Blood samples were evaluated for hepatic toxins and factor VII levels. RESULTS: Nine patients were enrolled (status I, n = 5; status IIA, n = 4). There was no significant change in MAP, HR, or Svo(2) (MAP: Pre = 81 +/- 5.6 mm Hg, Post = 79 +/- 5.9 mm Hg, P =.70; HR: Pre = 104 +/- 5.2 bpm, Post = 107 +/- 6.2 bpm, P =.62; Svo(2): Pre = 72 +/- 3.5, Post = 71 +/- 1.7, P =.77). There was a decrease in the ammonia and total bilirubin levels (NH(3): Pre = 129.8 +/- 23.8 mg/dL, Post = 63.9 +/- 16.1 mg/dL, P =.01; total bilirubin: Pre = 20.3 +/- 2.5 mg/dL, Post = 17.6 +/- 2.7 mg/dL, P =.4). There was a significant increase of the Fischer ratio and factor VII levels (Fischer ratio: Pre = 0.98 +/- 0.2, Post = 2.17 +/- 0.5, P =.038; factor VII: Pre = 13.9 +/- 4.9, Post = 23.2 +/- 4.8, P =.015). There was a significant decrease in the HES and ICP (HES: Pre = 3.8 +/- 0.1, Post = 2 +/- 0.7, P =.02; ICP: Pre = 37 +/- 3.9, Post = 13.3 +/- 2.8, P =.048). Of 5 status I patients, 1 recovered native hepatic function and 3 were bridged to transplantation. CONCLUSIONS: This phase I study suggests that albumin dialysis as a liver support device is safe and effective in clearing hepatic toxins, with an associated decrease in the HES and ICP. This encouraging efficacy data warrant further investigation with a phase II/III trial. PMID- 11490372 TI - Nuclear factor-kappa B is upregulated in colorectal cancer. AB - BACKGROUND: Chemoresistance may involve the anti-apoptotic transcriptional regulator, nuclear factor-kappa B (NF-kappa B). The purpose of this study was to determine whether chemotherapy induces NF-kappa B activation in a human colon cancer cell line (SW48) and whether NF-kappa B is constitutively activated in colorectal cancer. METHODS: SW48 cells were incubated with gemcitabine hydrochloride (Gemzar) in the presence and absence of the 26s proteasome inhibitor, MG132, and NF-kappa B binding (electrophoretic mobility shift assay), DNA synthesis (tritiated thymidine uptake), cell viability (3-[4,5 dimethylthiazol-2-yl]-diphenyl-tetrazolium bromide assay), and apoptosis (caspase 3 activity) were measured at 24 hours. NF-kappa B binding (electrophoretic mobility shift assay) was also assayed in 10 colorectal cancer tumors. RESULTS: SW48 cells demonstrated constitutive NF-kappa B binding that was enhanced by gemcitabine hydrochloride in a dose-dependent manner. MG132 inhibited NF-kappa B binding and enhanced gemcitabine hydrochloride's inhibition of DNA synthesis (gemcitabine hydrochloride = 73% +/- 1.4% vs gemcitabine hydrochloride + MG132 = 6% +/- 0.4%, P <.05), cell killing (gemcitabine hydrochloride = 87% +/- 2.0 vs gemcitabine hydrochloride + MG132 = 25% +/- 1.3%, P <.05), and caspase-3 activity (gemcitabine hydrochloride = 870 +/- 17.4 vs gemcitabine hydrochloride + MG132 = 1075 +/- 20.4, P <.05). NF-kappa B binding was increased in 8 of 10 colorectal cancer tumors compared with adjacent normal mucosa. CONCLUSIONS: Gemcitabine hydrochloride enhances NF-kappa B binding in a colorectal cancer cell line, whereas inhibition of NF-kappa B enhances gemcitabine hydrochloride's antitumor activity. NF-kappa B is also activated in human colorectal cancer. NF-kappa B may identify chemoresistant tumors, whereas inhibition of NF-kappa B may be a novel, biologically based therapy. (Surgery 2001;130:363-9). PMID- 11490373 TI - Predictors of health after operation for aortoiliac occlusive and aneurysmal disease. AB - BACKGROUND: We assessed the impact of comorbid conditions and revascularization for aortoiliac occlusive and aneurysmal disease and determined the functional health status of patients with the Medical Outcomes Study SF36 Health Survey. METHODS: One hundred twenty-five patients were surveyed prospectively, before operation, and at intervals ranging from 2 weeks to 2 years after operation. To identify the factors that influenced functional health, multiple regression analysis was performed to test the hypothesis that age, pulmonary disease, atherosclerotic heart disease, diabetes, aortoiliac occlusive disease (AOD) versus aneurysmal disease, and the preoperative physical summary score affected outcome. RESULTS: Regression analysis identified that before operation, the physical summary score (PCS) was affected by pulmonary disease, atherosclerotic heart disease, and AOD, and patients with AOD had significantly worse PCS than patients with aneurysmal disease (43.2 +/- 12.6 vs 30.1 +/- 8.3, P <.05). This difference was also present after 3 to 12 months, and the preoperative PCS was the strongest predictor of the postoperative score. For patients followed up between 1 and 2 years, there was no significant difference among the groups, and atherosclerotic heart disease and pulmonary disease were identified to most affect the PCS. CONCLUSIONS: Patients with AOD have significantly impaired physical function (as compared with patients with aneurysmal disease) that is successfully reversed with a surgical procedure. The functional health of patients after operation for aneurysmal disease returns to baseline after 3 and 12 months. Ultimately, cardiac and pulmonary comorbidities have a continued effect on the functional health of patients. PMID- 11490374 TI - Cyclic strain activates the pro-survival Akt protein kinase in bovine aortic smooth muscle cells. AB - BACKGROUND: Pulsatile pressure induced by the beating heart causes cyclic strain on arterial endothelial cells and smooth muscle cells (SMCs). This study examined whether Akt, a serine/threonine protein kinase known to promote cell survival by inhibiting apoptosis, is activated by cyclic strain in bovine aortic SMCs. METHODS: Bovine aortic SMCs were cultured on flexible-bottomed membranes and then serum-starved for 24 to 36 hours. The cells were then exposed to 150-mm Hg repetitive deformations, which created an average of 10% strain on the monolayer SMCs at a frequency of 60 cycles/minute for 0 (negative control) and 30 minutes. Platelet-derived growth factor (PDGF)--stimulated SMCs were used as positive controls. Phosphorylation of Akt was determined by means of Western blot analysis. An apoptosis assay (TUNEL) was also performed on SMCs exposed to cyclic strain. RESULTS: Akt phosphorylation was significantly increased over that of the negative control after 30 minutes of cyclic strain and in the PDGF group. Cyclic strain did not increase the prevalence of apoptosis in SMCs over the control. CONCLUSIONS: Cyclic strain activated the pro-survival Akt kinase. The pro survival function was supported by the fact that cyclic strain did not increase apoptosis in bovine aortic SMCs. This experiment suggests that cyclic strain may induce arterial wall thickening by tipping the balance toward arterial SMC proliferation through the inhibition of apoptosis. PMID- 11490375 TI - HER2 signaling--induced microvessel dismantling. AB - BACKGROUND: The human epidermal growth factor receptor 2 protein (HER2) signaling in breast cancer imparts a metastatic advantage to the cell, likely by regulating gene expression. The HER2 signaling up-regulates angiopoietin-2 (Ang-2), which disrupts endothelial cell (EC) adherens junctions. We postulated that HER2 signaling may facilitate angioinvasion by disrupting microvessel integrity. METHODS: Rat microvessels, embedded in collagen, were grown into capillary networks and cocultured with MCF-7 or HER2 overexpressing MCF-7 (HER) to test for microvessel breakdown. We quantitated this effect by determining the cumulative length of intact microvessels. Other experiments used Herceptin- or heregulin beta 1-pretreated MCF-7 cells to modulate HER2 signaling, or soluble Tie-2/Fc receptor fusion protein (sTie2) to sequester tumor-cell released Ang-2. RESULTS: The MCF-7 cells induced a time-dependent loss of microvessel integrity. At 12 hours, HER cells induced a 90% reduction in cumulative length (P <.05). Pretreatment with Herceptin reduced whereas heregulin beta 1 augmented microvessel dismantling (P <.01). Sequestration of Ang-2 significantly, though not dramatically, reduced the MCF-7 cell induction of microvessel dismantling (P <.01). CONCLUSIONS: We show that HER2 signaling in breast cancer cells leads to induction of microvessel dismantling, which may open a portal for angioinvasion. It appears that Ang-2 affects this mechanism, although other factors also function in microvessel dismantling. PMID- 11490376 TI - The effect of surgical site infections on outcomes and resource utilization after liver transplantation. AB - BACKGROUND: Although postoperative infections have a significant impact on morbidity and mortality after orthotopic liver transplantation (OLT), less is known about their economic implications. In this study, we sought to identify risk factors and estimate the impact of surgical site infections on 1-year mortality, graft survival, and resource utilization after OLT. METHODS: We studied 777 first, single-organ liver transplant recipients from the National Institute of Diabetes and Digestive and Kidney Diseases Liver Transplantation Database. Surgical site infections (n = 292, 37.8%) were defined as bacterial or fungal infections of the liver, intestine, biliary tract, surgical wound, or peritoneum within 1 year of transplantation. A subset of these (n = 159) occurred during the transplant hospitalization and were used to estimate excess charges associated with surgical site infections. RESULTS: Leaks in the choledochojejunostomy (odds ratio [OR] = 7.1, P =.001) and choledochocholedochostomy (OR = 2.5, P =.002), extended operation duration in hours (OR = 1.2, P =.002), serum albumin levels in grams per liters (OR = 0.71, P =.009), ascites (OR = 1.43, P =.037), and administration of OKT3 within 7 days (OR = 1.49, P =.039) significantly increased risk of infection. Surgical site infections did not significantly increase 1-year mortality (88.5% vs 91.5%, P =.19) but significantly increased 1-year graft loss (79.8% vs 86.5%, P =.022). Patients with surgical site infections incurred approximately 24 extra hospital days and $159,967 in excess charges (P =.0001). Multivariate analysis reduced the estimate of excess charges to $131,276 (P =.0001). CONCLUSIONS: Liver transplant recipients who develop surgical site infection have significantly higher resource utilization requirements than those who do not. These results imply substantial returns to preventative efforts directed at surgical site infections in patients undergoing OLT. PMID- 11490378 TI - Diamond stents for palliation of malignant bile duct obstruction: a prospective multicenter evaluation. AB - BACKGROUND AND STUDY AIMS: Various types of self-expandable metal stents have been introduced for biliary drainage in patients with malignant jaundice, showing prolonged patency compared with plastic endoprostheses. However, there has only been prolonged experience with a meaningful number of patients using the Wallstent. We evaluated the Diamond stent, a self-expanding uncoated biliary metal stent, in a prospective uncontrolled multicenter setting. PATIENTS AND METHODS: The eligibility criterion was obstructive jaundice due to inoperable malignant disease. Between August 1995 and January 2000, 126 patients, who received a total of 134 Diamond stents in four European centers, were followed prospectively. RESULTS: Technical and clinical success rates were 96 % and 98 %, respectively. No major procedure-related complications occurred. The 30-day mortality rate was 13 %. Stent occlusion occurred in 28 patients (22 %). Overall median stent patency was 477 days; overall median survival was 173 days. Stent occlusion, confirmed by endoscopic retrograde cholangiopancreatography, was successfully treated with plastic stents in all patients. Cost analysis revealed estimated costs of 3440 euros per patient for palliative treatment with the Diamond stent. CONCLUSIONS: The Diamond stent compares favorably with other biliary metal stents for patients requiring biliary drainage of malignant jaundice. PMID- 11490379 TI - Development of a sleeve sensor for measurement of sphincter of Oddi motility. AB - BACKGROUND AND STUDY AIMS: Unavoidable catheter movement during sphincter of Oddi (SO) manometry can produce considerable variations in the basal pressure, due to movement of the recording sidehole. The sleeve sensor is a perfused channel which records the highest pressure point along its length. The aim of the study was to develop and evaluate a prototype sleeve sensor for SO manometry. MATERIALS AND METHODS: Bench-testing was used to assess the dynamic performance of the sleeve and sidehole assemblies. Recordings were initially made with a standard triple lumen catheter and then with a purpose-built manometric assembly which had a 15 mm long sleeve sensor. RESULTS: A perfusion rate of 0.04 ml/min gave the best balance between baseline pressure offset and rise rate. Recordings were attempted in nine patients and successfully achieved in four. The sleeve and sidehole recordings of the maximal basal pressure did not differ significantly (mean +/- SEM, 86.1 +/- 26.5 mmHg vs. 90.1 +/- 21.0 mmHg, P = 0.57, r = 0.998). CONCLUSIONS: Unnecessarily high perfusion rates are being used for SO manometry. The sleeve sensor has the potential to monitor SO pressure more reliably than the currently used perfused sidehole method and should enhance the safety of prolonged SO manometry. PMID- 11490380 TI - Do we need the overtube for push-enteroscopy? AB - BACKGROUND AND AIMS: Push-enteroscopy is well established in many gastroenterological departments. There is no agreement about whether the use of an overtube is helpful to optimize the depth of insertion. In this prospective and randomized study we investigated the effect of the overtube for push-type enteroscopy. PATIENTS AND METHODS: Between August 1999 and August 2000, 80 patients (47 male, 33 female; age 63.1, range 20 - 90 years), who underwent investigation by push-enteroscopy were randomly allocated to two groups. Group T underwent investigation with the Olympus push-enteroscope SIF 100 with an overtube and group NT without an overtube. The length of insertion was estimated by fluoroscopy, by counting the folds of the small bowel and by insertion length in centimeters, while the scope was drawn back to the pylorus. The number of pathological findings was documented and patient comfort was measured by means of a standard questionnaire completed after enteroscopy. RESULTS: Both groups were comparable with regard to age, gender, indication for enteroscopy and pathological findings. There was a highly significant correlation between the length of insertion measured in centimeters and the counted folds in both groups (correlation 0.57, P < 0.001 in group T; 0.80, P < 0.001 in group NT). The length of insertion estimated by means of fluoroscopy was not reliable in determination of the exact length of insertion. There was a significant difference (P < 0.05) in the insertion depth between group T (mean values: insertion length 72.4 cm, counted folds 99.3) and group NT (mean values: insertion length 60.8 cm, counted folds 74.1). CONCLUSIONS: The use of an overtube in push-enteroscopy is advantageous with respect to the depth of insertion (significant difference between group T and NT in counted folds and the insertion depth in centimeters). At this time, this gain of insertion length did not result in a higher rate of pathological findings. Nevertheless, we would recommend the use of an overtube in performing push-type enteroscopy in order to increase the number of pathological findings. PMID- 11490381 TI - An assessment of the potential value of endoscopic ultrasound as a cost minimizing tool in dyspeptic patients with persistent symptoms. AB - STUDY AIMS: To quantify resource utilization in dyspeptic patients with persistent symptoms and to determine whether using both the endoscopic and ultrasound capabilities of endoscopic ultrasound could reduce costs. METHODS: Consecutive patients with persistent dyspepsia, after a minimum 1-month trial of acid suppression, underwent endoscopic ultrasound (EUS) and upper endoscopy using the GF-UM20 echo endoscope. Assuming EUS could replace imaging tests which had been requested in addition to upper endoscopy, the hypothetical costs of the EUS based and upper endoscopy-based strategies were compared. RESULTS: 116 patients with persistent dyspepsia underwent EUS, of whom 64.6 % had > or = 2 imaging procedures, most commonly computed tomography (CT) (70.6 %) and abdominal ultrasound (64.7 %). The number of tests did not correlate strongly with any demographic variables. The fiberoptic echo endoscope provided an adequate endoscopic and ultrasound examination but was damaged by retroflexion. Direct hospital costs were lowest for the EUS-based strategy. Total avoidable cost for 116 patients was $ 4137 to $ 14 121 (or $ 36 to $ 122 per patient), depending on whether upper endoscopy was performed in the non-EUS strategies. CONCLUSIONS: Patients with persistent dyspepsia may undergo multiple abdominal imaging procedures. Clinical variables do not predict the need for additional testing. An EUS-based strategy may reduce overall costs if it prevents additional testing. PMID- 11490382 TI - Potential impact of magnetic resonance cholangiopancreatography on endoscopic retrograde cholangiopancreatography workload and complication rate in patients referred because of abdominal pain. AB - BACKGROUND AND STUDY AIMS: Endoscopic retrograde cholangiopancreatography (ERCP) has a significant mortality, morbidity, and failed cannulation rate. Magnetic resonance cholangiopancreatography (MRCP) is a safer, noninvasive method of imaging the pancreaticobiliary tree. A substantial number of patients are referred for ERCP because of abdominal pain, a high proportion of whom have normal ducts or pathology not requiring interventional ERCP. The aim was to assess the potential impact of MRCP on overall ERCP workload and patient outcome if MRCP were the primary investigation in patients referred for ERCP because of abdominal pain. PATIENTS AND METHODS: 1758 consecutive ERCPs performed in 1148 patients over a 3-year period in a single tertiary referral center in the pre MRCP era were reviewed. Cannulation failure, ERCP findings, need for follow-up ERCP and all 30-day major complication rates were analyzed with regard to clinical indications. RESULTS: The overall workload comprised 1108 (63 %) successful initial ERCPs, 188 (11 %) failed cannulation attempts and 462 (26 %) follow-up ERCPs. Of the patients, 299 (27 %) had normal ERCP findings, 331 (30 %) had choledocholithiasis and 246 (22 %) had strictures. lf MRCP had been used as the primary imaging investigation in the 451 patients (39 %) referred for ERCP because of abdominal pain, we estimate that 197 patients (44 %) would have avoided ERCP, and the overall ERCP workload would have been reduced by 13 %. Initial MRCP in suspected gallstone pancreatitis and certain miscellaneous groups, it was estimated, would have further decreased ERCP workload by 9 %. Four of 40 major ERCP-related complications (3.5 %) and one of four ERCP-related deaths (0.35 %) would potentially have been avoided. CONCLUSIONS: Initial MRCP in patients referred with abdominal pain would potentially have avoided ERCP in 44 % of cases, reduced ERCP workload by 13 % and significantly reduced patient morbidity and mortality. The relatively small reduction in ERCP workload among these patients reflects the fact that over half of them had probable sphincter dysfunction, a significant proportion of whom might have benefited from biliary manometry and/or endoscopic intervention despite a normal MRCP. Furthermore, a small number of patients with calculi and subtle biliary and pancreatic strictures would be missed by this approach. PMID- 11490383 TI - Virtual CT cholangioscopy: comparison with fiberoptic cholangioscopy. AB - BACKGROUND AND STUDY AIMS: No studies comparing virtual computed tomography (CT) cholangioscopy of the common bile duct compared with fiberoptic cholangioscopy are available. The aim of our study was to evaluate the feasibility of virtual CT cholangioscopy of the common bile duct. PATIENTS AND METHODS: The study population comprised 52 patients (25 women, 27 men; mean age 56.5, range 32 - 81) with biliopancreatic disorders. Endoscopic images were produced by a volume rendering method and a perspective projection. The ability to detect the endoluminal view and abnormalities of the common bile duct by virtual CT cholangioscopy and fiberoptic cholangioscopy was evaluated. RESULTS: Except for two cases (4 %), virtual CT cholangioscopy revealed excellent and moderate endoluminal visualization. There was no significant difference between the techniques (virtual CT cholangioscopy vs. fiberoptic cholangioscopy: excellent, 73 % vs. 85 %, P = 0.149; moderate 23 % vs. 15 % (P = 0.319); poor, 4 % vs. 0 %, P = 0.153). Virtual CT cholangioscopy revealed no significantly different ability to detect stenosis and obstruction of the common bile duct, compared with fiberoptic cholangioscopy. However, the ability of virtual CT cholangioscopy to detect minute papillary tumors (virtual CT cholangioscopy 30 % vs. fiberoptic cholangioscopy 100 %, P = 0.001) and stones smaller than 5 mm (virtual CT cholangioscopy 25 % vs. fiberoptic cholangioscopy 100 %; P = 0.002 was significantly less than that of fiberoptic cholangioscopy. CONCLUSIONS: Virtual CT cholangioscopy cannot replace fiberoptic cholangioscopy completely. However, the use of this technique, instead of fiberoptic cholangioscopy, may be feasible for following up patients after biliary intervention. PMID- 11490384 TI - Endoscopic treatment for laterally spreading tumors in the colon. AB - BACKGROUND AND STUDY AIMS: Laterally spreading tumors (LST) of the colon are best removed by endoscopic mucosal resection (EMR) as they extend laterally rather than vertically. Since they sometimes invade deeply into the submucosal layer, it is important to assess the depth of invasion endoscopically before treatment. In the present study, we examined the endoscopic features of a large number of LSTs in order to assess which features correlated with depth of invasion. MATERIALS AND METHODS: 257 LSTs removed at the National Cancer Center Hospital, Tokyo, between January 1988 and September 1998 were retrospectively analyzed. RESULTS: With univariate analysis, unevenness of nodules, presence of large nodules, size, histological type, and presence of depression in the tumor were significantly associated with depth of invasion. Multivariate analysis revealed that histological type and depression in the tumor were independent factors predicting massive submucosal invasion. When an LST showed: 1) even nodules without depression, or 2) uneven nodules without depression and less than 3 mm in diameter, the risk of massive submucosal invasion was 0 % (0/121) and 3.7 % (3/82), respectively. CONCLUSION: When LSTs meet the above endoscopic criteria, EMR should be the first-line treatment because of the low risk of massive submucosal invasion. PMID- 11490385 TI - A new endoscopic tattooing technique for identifying the location of colonic lesions during laparoscopic surgery: a comparison with the conventional technique. AB - BACKGROUND AND STUDY AIMS: Knowledge of the exact location of colorectal lesions is necessary but difficult to establish during surgery. Thus, endoscopic tattooing has been used as an important preoperative marker for identification. Using the conventional technique, we injected tattooing agents directly into the colonic wall. However, to make sure that the tattooing agents were adequately injected into the submucosal layer, and to prevent spillage into the peritoneal cavity, we modified the conventional method and developed a new tattooing technique: using India ink with prior and subsequent injection of saline into the submucosa. The aim of this study was to retrospectively assess the clinical utility and potential complications of the above two techniques of endoscopic tattooing using India ink. PATIENTS AND METHODS: A total of 153 patients underwent laparoscopically assisted colectomy at the National Cancer Center Hospital East, Kashiwa, Chiba, Japan, between June 1994 and December 1999, and 91 patients underwent preoperative endoscopic tattooing by either the conventional or the new technique. The conventional and new techniques were used from June 1994 to December 1997 (n = 36) and from January 1998 to December 1999 (n = 55), respectively. Informed consent was obtained from all the patients. RESULTS: Using the conventional technique, the exact location of the lesion was identified in 31 of 36 cases (86.1 %) during surgery. The complications of this procedure were silent local peritonitis in two patients and reactive lymph node swelling in one patient (3/36, 8.3 %). Using the new technique, in 54 of 55 cases lesions (98 %) were easily identified (p = 0.034),and in only one there was a small leakage of India ink into the peritoneal cavity (1/55; 1.8 %), no other serious complications were observed. CONCLUSIONS: The results demonstrate that our new technique for endoscopic tattooing is probably better than the conventional technique for clinical use, in terms of diagnostic accuracy and safety, but this would have to be proven in a randomized comparison. PMID- 11490386 TI - Endoscopic management of foreign bodies in the upper gastrointestinal tract: report on a series of 414 adult patients. AB - BACKGROUND AND STUDY AIMS: Ingestion of foreign bodies is a common occurrence. Few papers in the literature report experience and outcome at tertiary centers. The aim of this paper is to report the management and the outcomes in 414 patients admitted for suspected ingestion of foreign body between May 1995 and December 1999. METHODS: A plain radiographic film of the neck, chest or abdomen was obtained in the case of radiopaque objects, and in order to rule out suspected perforation: in such cases a computed tomography (CT) study was also performed. All patients were asked to give their informed consent, which was refused by three patients. Anesthesia was always used, either conscious sedation (86.8 %), or general anesthesia in the case of poor patient tolerance (13.2 %). All patients underwent an endoscopic procedure within six hours of admission. A flexible scope was used in all patients and a wide range of endoscopic devices was employed. RESULTS: Foreign bodies were found in 64.5 % of our patients. Almost all were found in the esophagus. The types of foreign body were very different, but they were chiefly food boluses, bones or cartilages, dental prostheses or fish bones. In three patients (1.1 %) it was impossible to endoscopically remove the foreign body, which was located in the cervical esophagus: all these three patients required surgery. No complications relating to the endoscopic procedure were observed, but 30.7 % of patients had an underlying esophageal disease, such as a stricture. Only eight patients required a second endoscopic procedure, performed by a more experienced endoscopist. CONCLUSION: Foreign body ingestion represents a frequent reason for emergency endoscopy. The endoscopic procedure is a successful technique which allows the removal of the foreign bodies in almost all cases without significant complications. Surgery is rarely required. PMID- 11490387 TI - Thoracoscopic thermal ablation therapy for hepatocellular carcinoma located beneath the diaphragm. AB - BACKGROUND AND STUDY AIMS: Percutaneous interstitial thermal ablation therapy effectively treats hepatocellular carcinoma (HCC) that can be visualized on percutaneous ultrasonography. However, when the tumor is located just under the top of the diaphragm, visualization can be difficult with conventional ultrasonographic examination. There are also problems concerning complete tumor ablation. We performed thoracoscopic thermal ablation therapy for HCC located just beneath the diaphragm in nine patients with advanced liver cirrhosis. PATIENTS AND METHODS: Eight patients underwent thoracoscopic microwave coagulation therapy, and one patient underwent thoracoscopic radiofrequency ablation therapy. RESULTS: Despite the poor hepatic reserve, postoperative recovery after thoracoscopic thermal ablation therapy was rapid in all patients, without deterioration of hepatic function. CONCLUSIONS: This preliminary study suggests that the new technique of thoracoscopic thermal ablation therapy is a less invasive optional therapy for HCC located in segments VII or VIII in cirrhotic patients. PMID- 11490388 TI - Is there an ideal biliary metal stent? PMID- 11490389 TI - Endoscopic ultrasonography and magnetic resonance cholangiopancreatography in abdominal pain: what makes sense? PMID- 11490390 TI - Endoscopic mucosal resection. AB - We are now finding more malignancies in their early stages than previously. Attempts to ablate these lesions are difficult and do not provide the histological information required to decide on further treatment. Surgery is difficult to justify, as only a minority of lesions are associated with lymph node metastases and lesions may not become clinically relevant within the lifetime of an elderly patient. Endoscopic mucosal resection allows cancers to be resected at minimal cost, morbidity and mortality. It is also the most reliable investigation when assessing lesions which are suspicious for containing early cancer. After endoscopic removal, histological assessment of depth of penetration and a search for invasion into lymphatics or venules allows the risk of microscopic lymph node metastases to be predicted. The risk of developing metastatic disease can then be balanced against the risks of surgery in view of the patient's age and health. PMID- 11490391 TI - Transpapillary microwave coagulation therapy for recanalizing self-expandable metallic stents occluded by tumor ingrowth: initial experience. AB - Percutaneous microwave coagulation for recanalizing stents occluded by tumor ingrowth has been reported. With this technique, however, the percutaneous drain diminishes the quality of life in patients with unresectable tumors and a limited prognosis. Transpapillary microwave ablation was attempted in three patients with occluded stents. After a sheath had been inserted into the proximal hepatic duct across the occluded region, a microwave electrode was introduced into the intrahepatic duct via the sheath. We used microwave therapy with an output power of 40 W, based on our previous in vitro study. Except in one patient, the stents were successfully recanalized with one or two attempts. In one patient who underwent ablation in the intrahepatic duct, a 1.8-mm electrode enabled recanalization of the stent. In another who underwent ablation in the extrahepatic duct, however, a larger electrode was required. There were no procedure-related complications. Transpapillary microwave coagulation of occluded stents appears to be an alternative to percutaneous microwave coagulation with an electrode fitting the stent size. The technique might be easier with the use of a redesigned electrode with a guide wire lumen. PMID- 11490392 TI - Another novel use of endoscopic clipping: to anchor an esophageal endoprosthesis. AB - Endoscopic Hemoclips were developed as a method of securing permanent hemostasis in cases of nonvariceal gastrointestinal bleeding. However, Hemoclips have also been found useful in several other situations, such as closure of small perforations, as marking devices, and for anchoring feeding tubes. We describe here a further novel use of Hemoclips--to anchor an esophageal endoprosthesis. PMID- 11490393 TI - Duodenocolic fistula: an unusual sequela of stent migration. PMID- 11490394 TI - High-grade dysplasia in heterotopic gastric mucosa in the upper esophagus after radiotherapy: successful eradication 2 years after endoscopic treatment by argon plasma coagulation. PMID- 11490395 TI - Unusual cause of severe upper gastrointestinal hemorrhage treated using a simple endo-loop technique. PMID- 11490396 TI - Giant diverticulum of the ascending colon. PMID- 11490397 TI - Cytomegalovirus acute necrotizing esophagitis. PMID- 11490398 TI - Changes in periodontal therapy: evolution or revolution? PMID- 11490399 TI - Efficacy of implant placement after bone grafting for three-dimensional reconstruction of the posterior jaw. AB - A series of 21 men and 41 women received grafts resulting in mean faciolingual augmentation of 5.1 mm and mean vertical augmentation of 3.9 mm. In most patients, bone was obtained from the iliac crest. At the time of implant placement, the bone at the site was restored anatomically to type B in 50 patients and to type C in 12. The patients received from one to ten posterior implants, which were placed simultaneously with (n = 5) or approximately 6 months after grafting. All of the implants were loaded, with the follow-up ranging from 12 to 96 months (mean 37.3 months). Ceramometal restorations were ultimately used in all patients. Five patients suffered partial graft loss at a total of 21 implants, of which five (24%) failed. The total failure rate for implants placed in patients who received sinus + veneer grafts was 4% (9/222). In patients who received sinus and J grafts, the final implant failure rate was 2%. All implants placed in anteroposterior J grafts and mandibular grafts were successful. Overall, including replacement implants, the failure rate was 7% (23/329). Posterior implants can be placed after graft reconstruction with a success rate similar to that obtained without grafting, thereby improving the function and esthetic outcome. PMID- 11490400 TI - The clinical relevance of microbiologic testing: a comparative analysis of microbiologic samples secured from the same sites and cultured in two independent laboratories. AB - A field study using five different private periodontal practices was conducted; it compared two microbiologic culture samples simultaneously secured from the same sites within 23 individual patients and submitted for bacterial identification and antibiotic sensitivity testing to two separate laboratories. The results from the two laboratories were often different. In no instance did both laboratories agree on the presence of identical bacterial species. When only bacteria above threshold levels were compared, agreement was found in only nine of 23 cases. When examining antibiotic sensitivity, using 100% kill of all tested pathogens as the ideal, agreement between the two laboratories was poor. The laboratories agreed on the use of amoxicillin 17% of the time, tetracycline 26% of the time, and metronidazole 48% of the time. The use of amoxicillin and metronidazole in combination yielded a 78% agreement when the results of both laboratories were combined. It would appear from the data that the empirical use of amoxicillin-metronidazole combination therapy may be more clinically sound and cost effective than culturing and antibiotic selection based on the results of culture from any single microbiologic testing laboratory. PMID- 11490401 TI - Histologic evaluation of root coverage obtained with GTR in humans: a case report. AB - The purpose of this study was to histologically evaluate four teeth with recession defects that were treated with guided tissue regeneration (GTR). The amount of root coverage obtained ranged from 16.7% to 50.0%. In one of the defects treated, new bone was formed but none of the new bone was coronal to the original gingival margin. Additionally, in this one case new cementum and connective tissue attachment was formed. However, because the new bone, new cementum, and new connective tissue were not coronal to the original gingival margin, this result could not be classified as regeneration. The vast majority of the attachment in this defect was long junctional epithelial attachment in the portion of the recession defect covered by the root-coverage procedure. In the other three defects treated, there was a loss of bone. All of the root coverage obtained was a long junctional epithelial attachment in three of the four defects. The results of this study do not show regeneration in any of the four defects treated. PMID- 11490402 TI - Maxillary sinus floor augmentation in rabbits: a comparative histologic histomorphometric study between rhBMP-2 and autogenous bone. AB - The present study was designed to compare recombinant human bone morphogenetic protein-2 (rhBMP-2) impregnated on an absorbable collagen sponge (ACS) with particulate cancellous bone and marrow (PCBM) harvested from the iliac crest in an animal model of maxillary sinus floor augmentation. Bilateral sinus floor augmentation procedures were performed in 30 adult Japanese white rabbits. rhBMP 2/ACS or PCBM was grafted to each maxillary sinus. Animals were sacrificed at 2, 4, or 8 weeks after grafting, and bone formation in response to each implant was evaluated histologically and histometrically. Histometric results compared by analysis of variance revealed no statistically significant difference in the bone volume at augmented areas between the two types of implant (P > 0.05). Histologic evaluation documented that the trabeculae with a lamellar structure were embedded in fatty marrow at 8 weeks in both implant sites. These results suggest that sinus floor augmentation with rhBMP-2/ACS or PCBM induces comparable histologic and histometric evidence of bone formation in rabbits. PMID- 11490403 TI - Occlusal disease revisited: Part I--Function and parafunction. AB - This article will address the evolution of occlusal disease from childhood and the deciduous dentition forward. It will include the evolution of anterior wear with emphasis on "cross over" of the mandible. Having classified the types and extent of occlusal disease, new evidence will be shown, on the adult dentition with pathognomonic manifestations, of cross-over or bruxed-braced occlusal disease. Its potential effect on restorative dentistry and dental implants will be discussed. PMID- 11490404 TI - Occlusal disease revisited: Part II. AB - In part I of this article, the evolution of bruxism from childhood was discussed. Further, the different types of anterior tooth wear were reviewed. Specifically, the type of wear noted in bruxed-braced or cross-over position was pointed out. Examples were illustrated to allow the practitioner to recognize the type of parafunction in advance of treatment. This article will continue the discussion of cross over with moderate to extreme examples. Suggestions for treatment are discussed depending on the severity of the problem. Restorative failure and the implications for implant dentistry are noted. PMID- 11490405 TI - Plastic surgery at the time of membrane removal around mandibular endosseous implants: a modified technique for implant uncovering. AB - Removal of barrier membranes may complicate second-stage implant surgery, particularly in mandibular areas characterized by a shallow vestibule and minimal amount of keratinized tissue. A new surgical technique that permits implant exposure and membrane removal combined with a plastic procedure to improve soft tissue quality both buccally and lingually is presented. A midcrestal incision preserving the keratinized tissue available on the lingual side is designed. A double-layer flap is elevated, allowing membrane removal. The inner, full thickness layer is then sutured back into place, thus protecting the regenerated bone and allowing a recipient bed for a free gingival graft. The outer, partial thickness flap is sutured apically, thus deepening the vestibule. The advantages and technical aspects of the procedure are discussed. PMID- 11490406 TI - Alveolar ridge augmentation with Bio-Oss: a histologic study in humans. AB - The aim of the present study was to investigate the healing of alveolar ridge defects augmented with cancellous bovine bone mineral. In six partially edentulous patients, bone augmentation was necessary prior to implant placement because of severe alveolar ridge resorption. The defect sites, all located in the maxilla, were filled with Bio-Oss and covered with the resorbable collagen membrane Bio-Gide. Biopsies were obtained from the defect sites 6 to 7 months following grafting and were processed for ground sectioning. The histologic analysis revealed that the Bio-Oss particles occupied 31% of the total biopsy area. An intimate contact between woven bone and Bio-Oss was detected along 37% of the particle surfaces. A mixed type of bone was found; it contained woven bone and parallel-fibered bone, which demonstrates features of remodeling activity. Signs of resorption of the grafting material were observed in the histologic sections, which indicates that the material takes part in the remodeling process. It is suggested that Bio-Oss may be a very suitable material for staged localized ridge augmentation in humans. PMID- 11490407 TI - Root reshaping: an integral component of periodontal surgery. AB - It is the aim of this article to present a surgical option to the traditional method of returning lost biologic width where invasion of the junctional and/or connective tissue attachment associated with a tooth has occurred. The alternative to conventional osseous resection involves reshaping the existing tooth surface in combination with conservative removal of the supporting alveolar bone to create the width needed for the restoration to be biologically acceptable. This procedure accomplishes several goals: (1) minimum supporting bone is removed; (2) deleterious root surface anatomy, such as grooves, concavities, and cementoenamel projections, is diminished; (3) a smooth root surface that is more biologically acceptable to soft tissue is created; (4) Class I and II furcation lesions may be decreased or eliminated; and (5) improved gingival contours and space for restorative materials can be created in situations in which close root proximity is present. This article will present a step-by-step approach to using root reshaping as an alternative to traditional crown lengthening. PMID- 11490408 TI - [Congenital forms of a prolonged QT interval syndrome]. PMID- 11490409 TI - [Use of echocardiography with dobutamine for diagnosis of ischemic heart disease and evaluation of myocardial viability]. PMID- 11490410 TI - [Characteristics of arrhythmias and their treatment with diltiazem in patients with bronchial asthma]. AB - The authors' studies demonstrate that 87.5% of patients with exacerbation of moderate bronchial asthma have cardiac arrhythmias most of which have favorable prognosis. Most of the arrhythmias were supraventricular. Hypercatecholaminemia is considered one of the mechanisms contributing to development of arrhythmia in bronchial asthma exacerbation. Administration of diltiazem proved that this drug is highly effective in the treatment of both supraventricular and ventricular arrhythmias in asthma patients. PMID- 11490411 TI - [Inheritance of Wolff-Parkinson-White syndrome and evolution of its clinical symptoms in families of patients in a prospective trial]. AB - Medicogenetic examination was conducted in families of 46 patients (21 women and 25 men aged 16-74 years) with Wolff-Parkinson-White (WPW) syndrome. A total of 256 relatives were investigated (136 women and 120 men aged 2 to 85 years). As a result, the diagnosis of preexcitation syndrome and phenomenon was made initially in 75(29.3%) of the relatives: WPW syndrome, Clerc-Levy-Cristesco (CLC) syndrome, CLC phenomenon was made in 6(2.3%), 27(10.6%) and 42(16.4%) relatives, respectively. Additional conduction pathways in the families with WPW syndrome are inherited by the autosome-dominant type with penetrability 0.94(94%) and clinical polymorphism. Prospective observation of the families revealed evolution of the clinical symptoms (development of arrhythmia) in the relative with CLC or WPW phenomenon in unfavorable exo- and endogenic factors. WPW syndrome evolution in the patients ran with aggravation of arrhythmia though 12 patients showed improvement. PMID- 11490412 TI - [Regulation of arterial pressure in healthy subjects and hypertensive patients of old age]. AB - Central hemodynamics, microcirculation and vascular reactivity were studied in 61 patients with essential hypertension stage II and 64 healthy subjects at the age of 40 to 74 years. The process of aging is accompanied with elevation of arterial pressure and peripheral vascular resistance, left ventricular diastolic dysfunction, reduction of vascular sensitivity to noradrenaline and cold stimulation, deterioration of microcirculation and augmentation of oxygen consumption of the myocardium. PMID- 11490413 TI - [Effectiveness and side-effects of long-term treatment with glucocorticoids in patients with juvenile rheumatoid arthritis]. AB - The authors studied the response to and side effects of long-term glucocorticoid (GC) therapy of systemic juvenile rheumatoid arthritis (SJRA). GC were given to 24 girls and 9 boys at the age of 4 to 15 years with classic SJRA in a mean dose 1.6 +/- 0.13 mg/kg (24.0 +/- 0.55 mg/day) for 6.3 +/- 0.55 years, on the average. Mean duration of the disease was 7.1 +/- 0.55 years. 94% of the patients had lymphaticohypolastic constitution. Long-term GC monotherapy in a dose 0.2-0.5 mg/kg/day failed to control a SJRA course and to produce an immunosuppressive effect while side effects were severe: inhibition of activity of the hypothalamohypophysioadrenal system and resultant hormone dependence (100%), Cushing's syndrome (97%), nanism (85%), osteoporosis (64%), secondary infections (58%), aseptic necrosis of heads of the femur (52%). Oral GC treatment is not effective as first-line therapy of SJRA. It should be used as adjuvant to immunodepressants. Intraarticular and/or intravenous administration is preferable. PMID- 11490414 TI - [Acupuncture correction of autonomic nervous system disorders in patients with bronchial asthma]. AB - Acupuncture was added to medicines in the treatment of 192 patients suffering from bronchial asthma with autonomic dysfunction. The effect was assessed by changes in rhythmogram, external respiration function, psychological status. It was found that corporal acupuncture not only improves bronchial permeability but also reduces psychovegetative disorders. The effect of acupuncture was not related to that of placebo. PMID- 11490415 TI - [Non-narcotic analgesics and gastrointestinal lesions: risk factors, treatment, prevention]. PMID- 11490416 TI - [Pneumonia in patients with terminal renal failure on programmed hemodialysis]. AB - Of 183 patients with terminal renal failure (TRF) on programmed hemodialysis 12(6.5%) were diagnosed to have acute pneumonia. As a rule, the disease develops in hyperhydration and hypertension of the lesser circulation, more rarely concomitant heart disease is diagnosed which runs with general circulation insufficiency. This indicates that pneumonia in dialysis patients is secondary. Pulmonary congestion in pneumonia debut may mask infiltrative alterations. Therefore, in obscure cases it is desirable to combine x-ray examination with polyposition tomoscintigraphy of the lungs. The agent was verified in 33.3% cases. The sputum contained for the most part gram-positive bacteria. Antibacterial therapy was combined with measures against hyperhydration and hypertension. Mean duration of the treatment was 21.25 +/- 13.6 days. PMID- 11490417 TI - [Clinical-diagnostic significance of the study of medium mass molecular spectrum in patients with bronchial asthma combined with diabetes mellitus]. AB - Bronchial asthma (BA) and diabetes mellitus (DM) aggravate each other. Endogenic intoxication (EI) takes place both in BA and DM. Therefore, patients with BA and DM combination have chronic EI. A severe course of BA and DM can be judged not only clinically but also by spectrogram of medium mass molecules (MMM). BA patients have two peaks, maximal of them reaches 0.20 units of total area, total area being 0.4822 +/- 0.0186. In combination of BA with DM there are five peaks, on the average, maximal one reaches 0.30 uta and total area is 0.9207 +/- 0.029. Plasmapheresis, adjuvant to basic therapy, improves clinical course, MMM spectrogram exhibits less number of the peaks, their height and total area. PMID- 11490418 TI - [Antibacterial drugs in the treatment of osteoarthrosis: results of a prospective randomized trial]. AB - 65 patients with knee and hip joints osteoarthrosis with concomitant urinary infection and/or irritable colon syndrome were randomized into three groups: group 1 (21 patients) received doxicyclin 200 mg/day with nistatine for three weeks with 3-week interval between the courses and bifidum-bacterin administration between the courses and after the treatment; group 2 (22 patients) received biseptol 1920 mg for 3 weeks then 960 mg 10 days a month; group 3 (22 patients) consisted of controls who received no antibacterial drugs. The effect was assessed upon randomization, 3 weeks, 3, 6 and 12 months after the treatment. It was established that antibacterial treatment not only attenuated dysuria and dyspepsia but also had a significant positive effect on osteoarthrosis especially in patients with x-ray stage II, allowed to reduce doses of nonsteroid antiinflammatory drugs. Further studies will help specify the role of the infection factor in development of osteoarthrosis and potential of antibacterial treatment. PMID- 11490419 TI - [Aspects of development of spinal cord compression, neurological status and quality of life in patients with multi-organ tuberculosis]. AB - The authors study features of development of spinal cord compression, neurological affections and quality of life in patients with polyorganic tuberculosis. As shown by examination and treatment of 72 patients having pulmonary and spinal tuberculosis vs 46 control patients with spinal tuberculosis only, patients with pulmonary and spinal tuberculosis demonstrate faster development of spinal cord compression, neurological disorders, more severe and faster loss in motor activity and quality of life than patients with isolated spinal lesion. PMID- 11490420 TI - [Circadian rhythms of hemodynamic load on the cardiovascular system in patients with essential hypertension: implication for drug therapy rationale]. AB - Circadian rhythm of hemodynamic load by double product (heart rate x systolic blood pressure) was studied in 224 patients with essential hypertension (EH) treated with different antihypertensive drugs for 8-12 weeks and in 23 healthy subjects. The main feature of the rhythm was a marked morning rise in all the patients including non-dippers and night-peakers without nocturnal blood pressure fall. The examined drugs showed different effects on mean 24-h and morning values of the double product. PMID- 11490421 TI - [Chronological structure of arterial pressure in hypertensive patients on enalapril]. AB - 34 male patients with hypertension stage I and II aged 29-52 years (mean age 40.9 +/- 6.00) having mean 24-h arterial pressure (AP) above 135/85 mm Hg in mean daytime AP above 140/90 mm Hg and heart rate maximum 80 b/m entered the study of AP chronostructure in conditions of pure background and on enalapril treatment week 4, 8 and 12. The initial dose of the drug was 5 mg. Dose selection was controlled by 24-h AP monitoring. Enalapril was shown to significantly reduce mean daytime and 24-h AP as well as hyperbaric index, chronobiological time index, variability of systolic AP. The above dose selection brought more balanced AP lowering at daytime and at night. After 11 weeks of treatment no night-peakers were registered, the number of over-dippers decreased. Circadian rhythm of some hemodynamic parameters was characterized by a significant fall of rhythm average in unchanged acrophase and circadian AP amplitude indicating physiological action of enalapril. PMID- 11490423 TI - [A new blocker of proton pump pariet: pharmacological properties and effectiveness of clinical application]. PMID- 11490422 TI - [The role of heparin in prevention of deep veins thrombosis and pulmonary artery thromboembolism in therapeutic regime patients]. AB - The author reviews available literature on occurrence and effectiveness of prevention of deep vein thrombosis (DVT) and pulmonary artery thromboembolism (PAT) in therapeutic patients, MEDINOX trial, in particular. The latter trial has established that enoxaparine in a dose 40 mg/day for 6-14 days decreases the risk of DVT/PAT by 63% in inpatients. PMID- 11490424 TI - [Two cases of pachydermoperiostosis]. PMID- 11490425 TI - [Differential diagnosis of benign hyperbilirubinemia]. PMID- 11490426 TI - [Clinical pathogenic peculiarities of chronic Russian tick-born encephalitis]. AB - Clinical, serologic, epidemiological and neurological studies were carried out in 23 patients (18 men, 5 women) aged 15-69 years who had chronic Russian tick-borne encephalitis (RTE). With permanent stable humoral immunity the fact that these forms might develop was confirmed in 73.9% of the cases. In 26.1% of the patients were found to have incomplete vaccination and vaccine therapy combined with RTE viral infection. The following conditions underlay chronization were observed in all forms of acute RTE: fever in 4.3%, meningitis in 30.4%, focal lesion in 43.5% (the encephalitic syndrome in 8.7%, poliomyelitis in 8.7%, encephalopoliomyelitis in 26.1%). Initially progressive course was found in 21.7% of the cases. There was a primary rise of the hyperkinetic syndrome after the meningeal and focal forms of RTE with the encephalitic syndrome; while development of the amyotrophic syndrome (ATS) was found after the focal form with the poliomyelitic and encephalopoliomyelitic syndrome. The amyotrophic lateral sclerosis (ALS) syndrome occurs as both primary progressive RTE and ATS transformation independently of the initial RTE form. Latent periods of development of ATS and ALS syndrome were virtually equal (mean 1.4 years) and did not depend on the initial form of acute RTE. The duration of development of latent chronic forms makes it necessary to follow up RTE patients for at least 3 years. PMID- 11490427 TI - [Clinical characteristics of the endogenous intoxication syndrome in Russian tick born encephalitis]. AB - The paper characterizes the clinical and laboratory indices of the endogenous intoxication syndrome (EIS) in 173 patients with acute Russian tick-borne encephalitis. In its feverish form, EIS is characterized by the general intoxication syndrome and a slight increase in the luciferase index (LI). In its meningeal form, EIS is accompanied by overall cerebral symptoms, cardiovascular, respiratory and hepatic detoxifying dysfunctions, which are manifested by increases in both the intoxication leukocyte index of (ILI) and LI. In its focal form, EIS is characterized by the signs of multiorgan failure with the most pronounced alterations in the laboratory indices of endotoxemia (LID and LI). PMID- 11490428 TI - [Anticipation in the families of patients with alcoholism and drug addiction (son father dyads)]. AB - The clinical pattern of psychoactive drug addiction (PDA) was investigated in 41 father-son dads. Seven clinical characteristics were taken into consideration: The age at the onset of systematic PDA abuse; the rate of addiction development; the severity of the withdrawal syndrome; rate of progression, the form of drug abuse; the duration and quality of remissions; somatoneurologic consequences; and the clinical picture as a whole. These characteristics was evaluated in the members of each dad in terms of more or less severity. There is a strong tendency for more severe clinical picture in sons as compared with their fathers. The findings support the hypothesis that there is anticipation in the representatives of two generations of the families with drug dependence. PMID- 11490429 TI - [The role of clinical-electrophysiological indices in therapy for late-stage residual stroke by dynamic correction of proprioception]. AB - The new method dynamic proprioceptive correction using a medical loading costume was included into complex therapy in patients with late-stage residual stroke. Application of the costume promoted normalization of complex locomotor acts of walking by correcting proprioceptive pulsation resulted from the system of elastic draughts. Thus, a new motor stereotype was forced upon the patients. Clinical observation, computer analysis of the motor potential, spectrum of heart rhythm variabilities before and after therapy, and psychological testing were performed in 120 patients with motor disorders (pareses, paralyses) resulted from acute cerebral circulatory disorders. A steady-state clinical effect (p < 0.05) was found in 71.9% of the patients after 15 sessions of therapy. The patients became to walk independently, a possibility to self-attendance appeared, their speech became better. Lateralization of a damage and preservation of both the most significant homeostatic vasomotor mechanisms and the cerebral mechanisms of preparation of a movement with the minimal manifestations of emotional-affective disorders were shown to have impact on the patients' rehabilitation and on the early clinical features of pyramidal defect. PMID- 11490430 TI - [Balneotherapy of children with sequelae of brain injury]. AB - The paper presents manifestations of aftereffects of cranio-cerebral trauma in children. A lot of clinical observations grounded the expediency of a combined sanatorium treatment of such patients using mineral baths of different chemical composition. Radon baths are preferable, especially in patients with hypertensive hydrocephalus syndrome. PMID- 11490431 TI - [Design basis of randomized clinical trial: practical approach]. PMID- 11490432 TI - [Proton magnetic-resonance spectroscopy in remitting and secondary-progressive multiple sclerosis]. AB - Proton magnetic-resonance spectroscopy (PMRS) was used to measure the levels of inositol/myoinositol (Ins), choline, creatine/phosphocreatine (Cr), glutamine/glutamate (Glx/Glx1), N-acetylaspartate (NAA), gamma-aminobutyric acid (GABA) and lipids were measured in the foci of demyelinization in the brains of 59 patients with multiple sclerosis (MS). Magnetic-resonance imaging was performed during a single investigation. A control group comprised 20 healthy individuals. PMRS revealed significant alterations in the levels of metabolite in all the patients as compared with the controls: decreases in NAA by 23-52%, in Cr by 12-21%, in choline by 15-26%; increases in Ins by 51-63%; as well as the appearance of lipids (up to 100%). In MS, there were reduction in NAA/Cr, NAA/choline, and NAA/choline/Cr ratios by 12-53; 10-19; and 57-82%, respectively. As compared with the remitting MS, secondary-progressive MS showed decreases in the content of NAA by 23-25%, NAA/(choline + Cr) by 48-54% and increases in the levels of Ins and lipids by 50-76%. In remitting MS, there was a strong correlation between the NAA/Cr ratio and the volume brain lesion. It is concluded that PMRS evaluated the extent, pattern and activity of demyelinization (by the levels of Ins, NAA, Cr, lipids) and the intensity of cerebral atrophy (by NAA levels, NAA/Cr ratio). The findings testify that there are neurochemical differences between remitting and secondary-progressive MS. PMID- 11490433 TI - [Personality disorders: old and new problems]. AB - Correlations were found between the structure of personality disorders (PD) (neurotic--borderline personality organization) and the regularities of PD dynamics. Taking into consideration the regularities of interaction of psychopathologic manifestations with positive symptomatology, three types of PD dynamics were psychopathologically identified, which show information on polar (compensation--decompensation) processes: 1) that in form of personality development; 2) that in form of PD-associated quasipsychoses; associated with DP; 3) that in form of transformation of characterological disorders into psychopathologic ones. The first two types of dynamics, which appear as as actualization of pathocharacterological formations in the pattern of mental disorders, reflect decompensation processes. In neurotic personality (emotionally labile, anxious, dependent, sensitive schizoids, etc) there is a deep (in terms of personality development) reorganization of characterological abnormalies. In borderline personality (paranoid, expansive schizoids, anancasts, borderline, etc.), aggravation and chronic pattern of mental disorders occurs with dynamics of PD-associated quasipsychoses. The third type of dynamics is not accompanied by aggravation of pathocharacterological manifestations and reflects compensation process. Separation of the isolated symptom complexes, PD derivatives (steady state catathymically charged pathocharacterological formations are symptom forming) is possible in any type of personality organization, but more frequently is observed at complex forms with a high proportion of facultative complexes and accentuations. PMID- 11490434 TI - [Psychological aspect of psychosomatic pathology of the colon]. AB - Taking into consideration the age of individuals, 100 patients (25 men and 75 women) aged 21-60 years were examined by using clinical, psychological and statistic (correlation) analysis to determine whether there are relations between clinical manifestations of the irritable bowel syndrome and personality. They were all referred to a psychiatrist from a gastroenterology hospital. The formation and dynamics of this syndrome, as well as its the clinical variations (diarrhea, pain, constipation) are largely determined by personality traits (increased anxiety; high inclination to somatization of emotions with inability to adequately verbalize them, pronounced intrapunitivity; direction of hostility to themselves; permanent inclination to depressive reactivity) and a passive defensive behavior under the conditions of frustration. PMID- 11490435 TI - [Analysis of mutations in ATP7B gene and experience with direct DNA-diagnosis in hepato-lenticular degeneration]. AB - Hepatolenticular degeneration (HLD) is a severe autosomal-recessive disorder of the copper metabolism. It is characterized by excessive accumulation of copper in the brain and in viscera and is conditioned by the damage in the gene of copper ATP-ase (ATP7B). The paper presents the results of screening of ATP7B gene mutation in 42 patients with HLD from Russian population. The regions of ATP7B gene that are the most frequently exposed to the mutation have been studied (the exzones 14, 15, 16, 18). It is demonstrated that A-->C mutation in the 14-th exzone that led to the change of histidine1069 amino acid for glutamine, was found in more than 60% of patients--Slavs from the European Russia. This mutation was observed in both homo- and heterozygous states. The deletion of (CCC-->CC) nucleotide in the 15-th exzone of the gene was observed in 2 cases. The detailed analysis of the clinical-genetic correlations was performed in patients with the determined damages of ATP7B gene. In Russia the experience of the direct DNA diagnosis of HLD is described for the first time. It is significant for early evaluation of the patients in preclinical state and for prescription of the preventive copper-eliminating therapy. PMID- 11490437 TI - [The instrumental method in the investigation of the muscular tone]. PMID- 11490436 TI - [Molecular genetic polymorphism of the genes of neurotransmitter systems in schizophrenics with early manifestation of the disease]. AB - Molecular-genetic polymorphism of genes-candidates was investigated: the genes of serotonin receptor--type 2a (HTR2A), dopamine receptor gene--type 2, serotonin transporter (5HTTLPR). Thirty one schizophrenic patients whose age was 12.6 +/- 3.6 years at the onset of the disease and 208 patients whose age was 23.5 +/- 6.7 years at the onset of the disease were examined. The frequencies of HTTLPR and DRD2 genotypes differed insignificantly in both groups. The distribution of 5HTR2A genotypes in the schizophrenic group with an early manifestation of the disease differed from that with a later manifestation significantly (chi 2 = 6.27; df = 2; p = 0.044). The relative risk (odds ratios) was 7.9 with 95% significance interval 1.008-61.94; p = 0.045. The severity of the disease and a positive family history were also examined in A2A2 genotype carriers. A positive family history was found in 9 (52.9%) of the 17 schizophrenics with an early manifestation and only in 15 (21.1%) of 71 patients of the similar group with a later one. Assessment of the clinical symptoms revealed that the total scores by the negative symptomatology subscale (PANSS) was higher in the patients with an early manifestation than in those with later one; but these differences did not achieve the significance level. These and earlier findings lead to the conclusion that A2A2 genotype was more frequently observed in the patients with more pronounced negative symptoms and high hereditary burden, which suggests that the A2A2 genotype is associated with an early onset. PMID- 11490438 TI - [Dynamics of the aged out-patients with mental disorders]. PMID- 11490439 TI - [The role of the cerebellum in the organization of higher mental functions]. PMID- 11490440 TI - [Magnetic resonance imaging in the diagnosis of multiple sclerosis]. PMID- 11490441 TI - [WHO/EURO databases on the trends of the development of mental health services (1992). The address by V.S. Iastrebov]. PMID- 11490442 TI - [Neuropsychological deficit in chronic heroin abusers]. AB - Neurological consequences of chronic heroin exposure are poorly known. 38 male patients with current heroin abuse or dependence were examined in withdrawal period that lasted more than 10 days, and were compared with 19 healthy controls. Wisconsin Card Sorting Test (WCST), "Delayed Alternation" Test (DAT), "Tower of London" Test (TLT), Russian version of WAIS (1995) were used. Patients were medicated and medication status was evaluated by psychomotor speed level. Patients with the duration of daily heroin abuse more than 1.5 years performed significantly less effectively TLT solutions as compared with the healthy controls (after Bonferroni correction, p = 0001). Patients with shorter duration of daily heroin abuse had a trend to perform TLT solutions poorer as compared to healthy controls and better than group with longer duration (after Bonferroni correction, p = 0.07 and 0.08). Three groups did not differ by WCST and DAT significantly, and general intelligence was in normal range in three groups. Multiple regression analysis confirmed significant influence of daily heroin abuse duration on TLT performance efficiency in our population (beta = -0.426, p < 0.05) without effect of age, education, IQ, dosage of heroin per day, withdrawal duration and current medication status (psychomotor speed level). Perseverative responses on DAT were significantly related to daily heroin dosages before treatment (beta = 0.405, p < 0.05) and negatively correlated with the withdrawal duration. These data give grounds to suppose, that chronic heroin exposure impairs planning functions of prefrontal cortex (TLT), that can be explained by cumulative neuronal damages of prefrontal cortex and VTA dopamine neurons. That was demonstrated in experimental and morphological studies of opiate addicts who died after opiate overdose. Large doses of heroin can induce more extensive functional impairment with possible involvement of orbit frontal cortex. The latter deficit may be partially reversible during short-term withdrawal. PMID- 11490443 TI - [The disturbances of sensitivity in women with opium addiction]. AB - 52 women with opium addiction in remission (deprivation as a result of imprisonment) were examined. The disorders of appreciation pain were found in 50 cases (96 +/- 3%). There were clinical peculiarities of their topography (composed distal, maculary-mosaic and perioral-axial versions). These peculiarities were shown in alcoholism, neuroses, neurosis-like nonpsychogenic conditions of infection and traumatic genesis. The supposed mechanisms of described disorders are also presented. PMID- 11490444 TI - [Optimizing the therapy of symptomatic partial epilepsy with sustained release carbamazepine]. AB - This article summarizes a three-year experience of treating 34 patients with carbamazepine (CMZ) retard with partial epilepsy who had previously been treated with a standard form of CMZ. The following steps were taken in order to optimize the therapy: the usage of monotherapy with CMZ, an increase of CMZ dose if it was necessary, usage of CMZ retard-form. Increase of the dose (more than 1400 mg) resulted in adverse reactions in 6 patients, which regressed when the patients started to be treated with CMZ retard form. On the whole, optimization of therapy provided the following effects: 18 patients (53%) did not experience any seizures, in 11 patients (32%) a frequency of the seizures was considerably decreased (by 50% and more as compared with the baseline level). Individual analysis of the seizures frequency showed that in patients treated with CMZ retard-form the maximal decrease of this frequency was observed for sleep seizures. A frequency of seizures decreased by 63% at night and by 34% in day hours. PMID- 11490445 TI - [Modern psychiatric service and criteria for the estimation of its activity]. AB - During the last decade the efforts of psychiatrists and managers of psychiatric service were directed to solving the problem of modern system of psychiatric care and its quality. In Russia the problem has to be solved under conditions of both social instability and economic crisis. The paper presents the data concerning investigation of the problem in terms of epidemiological, social, economic, ethical-juridical and organization criteria. The author concluded that the significant criteria for efficiency of the psychiatric service activity may be the following: satisfaction of the patients with this service; quality of their life; the degree of social integration; the level of their stigmatization in the society; the data about consequences of mental diseases for the family and the society; scientific-founded standards of medical-diagnostic and medical social aid. Social support and care of mentally ill patients and members of their families are essential in modern psychiatry. PMID- 11490446 TI - [Dynamics and prognosis of mental disorders incidence in elderly patients]. AB - Mental disorders incidence grows constantly in patients of 60 and older. It has grown by 9.8% in 1998 as compared with 1993 and by 21.2% as compared with 1991. In 1998 psychoses and mental retardation incidence was 117.1, while nonpsychotic forms of the disorders--136.4 per 100 thousands of population at the age of 60 and older. The main increase of the incidence was found in the group with disorders characteristic for the late age. A number of the new patients with senile psychoses and dementias increased in 1998 by 21.3% as compared with 1993, while a number of patients with nonpsychotic disorders of vascular nature during the same period increased by 61.2% and with the disorders of the other organic nature--by 35.3%. From 1995 the individuals of the old age had high incidence of reactive states--by 58.9%, of neuroses--by 71.8% and of psychosomatic diseases- by 72.2%. The paper predicts further increase of mental disorders incidence in the population of elderly and old age. PMID- 11490447 TI - [Contemporary system of mental health care: epidemiologic aspects]. AB - The authors suppose that adequate mental health care management should be based on the knowledge about the number of its potential patients. In order to determine this number the authors formulated a hypothesis that the necessary period of the observation of patients depends on the course of the disease and on the risk of the relapse. From the authors' point of view all mental disorders comprise 3 groups. Patients who belong to the 1-st group need observation during all their life; patients from the 2-nd group need observation during an active period of the illness; patients from the 3-d group need observation during the acute period only. On the basis of estimation of the full number of mentally ill people in Russia the authors calculate a size of each group and the total number of the potential patients--about 21 million of people (14% of the Russian population). This rate has been defined as "the actual number of mental patients". PMID- 11490448 TI - [Pathogenic pharmacotherapy in the acute phase of the withdrawal syndrome in opium addiction]. PMID- 11490449 TI - [Aspects of clinical manifestations of the disease in right and left-sided stroke]. PMID- 11490450 TI - [The experience of the work of social-psychological school for the relatives of mentally ill patients]. PMID- 11490451 TI - [Genetics of amyotrophic lateral sclerosis]. PMID- 11490452 TI - [Protection of mental health: a priority task of the state and the society]. AB - The paper is devoted to the World Health Day 2001--Mental Health Day. It gives a history of World Health Day. The state of the psychiatric science in Russia, the reforms carried out in psychiatry during the last decade and some measures of the perspective development of Russian psychiatry, proposed by the scientists and by the heads of some regional psychiatric services during the last years are considered. PMID- 11490453 TI - [Formal expert estimation of the quality of the research publications in psychiatry]. PMID- 11490454 TI - [Very high frequency electromagnetic irradiation in multimodal treatment of patients with disseminated infiltrative pulmonary tuberculosis]. AB - Multimodality treatment involving very high-frequency electromagnetic radiation (VHFER) in combination with the antioxidants alpha-tocopherol and sodium thiosulfate, which had been performed in 27 patients with disseminated infiltrative pulmonary tuberculosis, was effective. As compared to patients receiving chemotherapy in combination with electromagnetic radiation (n = 29) and routine chemotherapy alone (n = 29), these patients had more benefits from the multimodality treatment in terms of bacterial isolation cessation and reduced hospital stay by 1.5-2 months, minimal pneumofibrotic changes occurred in 63% of the patients. Combined VHFER and antioxidative therapy were found to exert a normalizing effect on lipid peroxidation and immunity. PMID- 11490455 TI - [Pathogenetic mechanisms of impaired intestinal microflora in patients with destructive pulmonary tuberculosis]. AB - Following 2-3 months of combined chemotherapy, most (97.5%) patients with destructive pulmonary tuberculosis were found to develop pathological changes in the intestinal microflora. Of them 92.3% had bifidobacterium deficiency, which creates conditions for opportunistic microorganisms to colonize the bowel, promoting the development of an inflammatory process in the gastroduodenal area, Bifidobacterium deficiency being a pathogenetic factor and opportunistic microbes acting as an etiological factor of the impaired intestinal microflora. Assessing of dysbiotic impairments makes it possible to specify the mechanisms of development of toxic reactions and inflammatory processes in the digestive tract. PMID- 11490456 TI - [Differentiated approach to preventive treatment of new cases of tuberculosis in children]. AB - The efficiency of various preventive treatment regimens was studied in 330 new cases of childhood tuberculosis by using clinical and laboratory findings and follow-up data. The paper shows it expedient to conduct a comprehensive immunological studies by tuberculin diagnosis and to examine immunity both at the time of infection detection in children and at their follow-ups. Monotherapy by the routine regimen can be performed in low tuberculin-sensitive children without risk factors. Children who have a complex of risk factors need long-term preventive treatment with two tuberculosis controlling agents. When immune responsiveness declines in children, it is necessary to apply pathogenetic agents that possess an immunomodulating effect, which substantially raise the efficiency of preventive treatment. PMID- 11490457 TI - [Components of monitoring drug resistance of tuberculosis agent in the evaluation of effectiveness of the national tuberculosis control program]. AB - The authors analyzed the changes and pattern of drug-resistance in tuberculosis pathogen by using the reports of over 700 microbiological laboratories of tuberculosis control service in Russia in the past 20 years (1979-1998), which yielded a monitoring system for drug resistance in the pathogen in this country. Seven underlying informational blocks were identified. These included more than 20 different quantitative and qualitative parameters covering the characteristics of a patient and the pathogen and regionally environmental and epidemiological indices. Analysis of the data on the united northern, west-northern, and Baltic regions is given as an example of successful use of the monitoring system. PMID- 11490458 TI - [Recognition of lymphatic tumors of the mediastinum]. PMID- 11490459 TI - [Surgical treatment of pulmonary tuberculosis: experience and prospects]. AB - The author's experience accumulated over 40 years shows that two major forms of surgical pulmonary tuberculosis (restrictive and disseminated) should be distinguished. The treatment of the former patients is highly effective and safe. To treat patients with progressive disseminated tuberculosis, especially in those who isolate Mycobacteria resistant to previously used drugs is difficult and expensive, which requires non only new generation antituberculous drugs, but comprehensive complex preparation for surgery. The author prefers collapse surgical interventions, but, if necessary, resorts even to pulmonectomy and application of an artificial pneumothorax contralaterally. Lobectomy is a major type of resection of the lung. Thus, surgery as a constituent of ensures recovery (stabilization of the process) in the most critically ill patients with pulmonary tuberculosis. PMID- 11490460 TI - [Complex radiologic diagnosis of renal tuberculosis]. AB - The paper analyzes a complex of physical, laboratory, and radiation studies in 71 patients with renal tuberculosis verified by the bacteriological urine test for Mycobacterium tuberculosis (MT) or postoperative histology. Ultrasound study, computed tomography, and excretory urotomography are not shown to be of informative value in diagnosing parenchymatous tuberculosis of the kidney. The diagnosis of this form of tuberculosis is made only on its clinical manifestations and positive urinary MT test. In all other forms of renal tuberculosis and its sequels, pathological changes can be detected on the basis of complex radiation techniques. PMID- 11490461 TI - [Ex juvantibus therapy in the differential diagnosis of urogenital tuberculosis]. AB - To make a differential diagnosis, 274 patients were followed up. Of them 68 patients were further found to have urinary tuberculosis that was verified by hypodermic tuberculin test, 15 patients underwent tuberculin diagnosis without special preparation, and 20 required type 1 ex juvantibus therapy. Challenge treatment with low-spectrum antituberculous agents in usual doses for 2 months confirmed the diagnosis in 3 patients and excluded it in other 3. PMID- 11490462 TI - [Features of hysterosalpingogram in verified genital tuberculosis]. AB - Two hundred and fifty one women were examined. Of them, 93 patients with active tuberculosis made up Group 1, 38 with inactive tuberculosis Group 2, and 120 with nonspecific gynecological diseases Group 3. In All tuberculosis was verified by morphological and bacteriological studies. Analyzing the X-ray pattern demonstrated that their leading X-ray signs were rigidity and clavity deformity of the uterine tubes and contrast shadows in the projection of uterine adnexa, and obliteration of the uterus. These signs were observed in 63.2% of the patients with inactive or long-lasting tuberculosis. At the same time young women with active tuberculosis firstly detected had no pronounced anatomic changes, which makes it possible to be initiate treatment timely and preserve reproductive function. PMID- 11490463 TI - [Effects of plasmapheresis with transfusion solutions on blood biochemical parameters and central hemodynamics in patients with chronic obstructive bronchitis]. AB - The impact of plasmapheresis (P) using 0.9% physiological solutions of sodium chloride solution and 5% lactoprotein on some blood biochemical and central hemodynamic parameters was studied in 20 patients with chronic obstructive bronchitis. No matter what hemocorrector was used, P was found to promote reductions in blood medium-weight molecules of peptides, overall blood proteolytic activity, alpha 1-proteinase inhibitor, ceruloplasmin, transferrin and to increase lipid peroxidation. At the same time curative P using lactoprotein favours better central hemodynamics, which indicates its ability to correct postexfusion plasma volume when used in a dosage of 10 ml/kg body weight). PMID- 11490464 TI - [Intrapleural hemorrhage after lung surgery]. PMID- 11490465 TI - [Significance of bronchospasm in the development of bronchial obstruction in lung diseases]. AB - VC, FVC, FEV1/VC%, PEF, MEF25, MEF50, MEF75 were studied in 254 patients with lung diseases with bronchial obstruction before and after berotec and antrovent inhalation. Bronchospasm was detected in most patients with bronchial asthma, in a half with exogenous allergic alveolitis, in more than a third with chronic bronchitis and pulmonary tuberculosis, and in a fifth with pulmonary sarcoidosis. In most examinees with bronchospasm, the proportion of the latter in bronchial obstruction amounted to 61-100%. PMID- 11490466 TI - [History, traditions and the present day]. PMID- 11490467 TI - [Morphological features of tuberculosis of the central nervous system]. AB - To study extrapulmonary tuberculosis, the authors analyzed deaths from that of the meninges and central nervous system over 5 years (1993-1997). The clinical and morphological variants of the disease, age-sex pattern, and the specific features of its course and outcomes are described. PMID- 11490468 TI - [Causes of death of patients with tuberculosis]. AB - The authors analyzed 310 case histories of patients who had died from tuberculosis. The factors predisposing to death were shown to be poor social being, bad habits, severe tuberculosis, and concomitant diseases. Among them there were prevalent fibrocavernous or disseminated tuberculosis that was complicated by the development of caseous pneumonia in every four patients. The main causes of death were progressive tuberculosis (52.7%), cardiopulmonary failure (26.9%). Generalized tuberculosis was responsible for death in 6% of the patients. PMID- 11490469 TI - [Causes of death of patients admitted to the tuberculosis dispensary from the pulmonological units]. AB - At present, pulmonological units admit patients with pulmonary tuberculosis forms that are epidemiologically hazardous, such as disseminated caseous and fibrocavernous pneumonia. The forms are characterized by abundant bacterial isolation and multiple bilateral lung tissue destruction. The causes of death in new cases of tuberculosis were their late referral for medical aid in the period of drastic deterioration of their health due to progression. Nineteen (63.3%) patients were found to have a gradual onset detected while thoroughly obtaining history data. Salivary MT tests were performed only in 50% of the patients admitted to pulmonological units, which makes an error diagnosis of acute pneumonia and unjustifiable treatment with broad-spectrum antibiotics. PMID- 11490470 TI - [Somatic diseases as cause of death of patients with active tuberculosis]. AB - Two hundred and fifteen case histories were studied to clarify causes of death in patients with active tuberculosis who had died from it. Males accounted for the largest proportion (86%) (n = 185). Patients above 50 years were 63.3% (n = 136). Two thirds were patients with restrictive tuberculosis without destruction and bacterial isolation. A hundred and fifty (70%) and 30% patients died from somatic diseases and other causes (sequels to alcohol abuse, violent death, suicide, etc.), respectively. Of the somatic diseases, cardiovascular diseases head the list, cancer ranks next to it, nonspecific respiratory diseases occupy the third place. PMID- 11490471 TI - [Fatal outcomes of caseous pneumonia in alcohol abusers]. AB - In alcoholics, caseous pneumonia is characterized by a malignant progressive course and therapy resistance. The toxic allergic syndrome in such patients may be a diagnostic sign of transition of benign forms of tuberculosis to caseous pneumonia. PMID- 11490472 TI - [Case of calculous pancreatitis in a patient with active form of pulmonary tuberculosis]. PMID- 11490473 TI - [A case of tertiary syphilis of the lungs and spine masking as tuberculosis]. PMID- 11490474 TI - [Formation of multiple tuberculomas in the lung with cystic hypoplasia]. PMID- 11490475 TI - [Pleural mesotheliomas]. PMID- 11490476 TI - [Radiation diagnosis of respiratory sarcoidosis]. PMID- 11490477 TI - [Tuberculosis in epidemics of HIV/ AIDS and drug addiction]. PMID- 11490479 TI - [Development of tuberculosis control system in the Sumsk region at the beginning of the 19th century]. PMID- 11490478 TI - [Principles and features of teaching phthisis specialty at the Department of Pediatrics of medical higher education establishments]. PMID- 11490480 TI - [Interregional Scientific-Practical Conference on Tuberculosis Control in the Far North: results, problems, prospects (Yakutia)]. PMID- 11490481 TI - [International Conference on Current Aspects of Thoracic Surgery]. PMID- 11490482 TI - [Improved technology of chemotherapy of pulmonary tuberculosis complicated by other diseases]. AB - Ample clinical material obtained in Yakutia was used to study the efficiency of combined treatment in patients with disseminated pulmonary tuberculosis concurrent with gastrointestinal and respiratory diseases. At the same time it was justifiable to employ alternatives to deliver antituberculous agents to the lesion focus (rectal dropwise administration, phono- and photophoresis of isoniazid, endobronchial colloid diluted rifampicin, and activated silver water diluted antituberculous drugs) with additional biophysical exposure. This made this group of patients have much better antituberculous agent intolerance, which increased nonbacterial isolation rates and closed decay cavities as compared with conventional methods for administering antituberculous agents. PMID- 11490483 TI - [Percutaneous laser-induced thermotherapy of nodular goiter]. AB - Substantiation of use of laser radiation Nd:YAG (1064 nm) and diode (805 nm) for destruction of thyroid tissue was the purpose of this study based on experimental research of reparative and adaptive processes in thyroid gland. The influence of high-intensity laser radiation was studied in vivo in thyroid gland of 41 dogs. The optimal modes of laser radiation ensured focal destruction of thyroid gland without damage of surrounding organs were selected. The developed modes were used in the treatment of 18 patients with recurrent nodal and multinodal euthyroid goiter. Exposure was performed by puncture method without denudation of thyroid gland. There were no complications during irradiation and nearest postoperative period. In dynamic ultrasonic examination over 2 years the decrease of nodes sizes was revealed in all the patients. PMID- 11490484 TI - [Combined use of low-frequency ultrasound and photodynamic therapy in the prevention of postoperative pleural empyema]. AB - Results of surgical treatment of 20 patients with malignant lung tumors operated in N.N. Burdenko faculty surgical clinic were analyzed. The authors have developed the method of prophylaxis of acute postoperative pleural empyemas patented in Russian Federation. This method is: after main stage of operation (lob- or pneumonectomy) pleural cavity in filled with antiseptic solutions (furacillin 1:5000, 0.02% chlorgexidin). 5 ml of photosense (sulfured ftalocyanin of aluminium) were injected in pleural cavity, after it pleural cavity is treated by low-frequency ultrasound. After it the antiseptic is removed and pleural cavity is irradiated by red light with use of KAMIN-VIDEO lamp. In all the 20 operated patients who had undergone sanation of pleural cavity by low-frequency ultrasound and it irradiation by KAMIN-VIDEO lamp the postoperative period was uncomplicated. PMID- 11490485 TI - [Proximal resection of the cardiac portion of the stomach]. AB - From 1989 to 1999 75 proximal resections of the stomach were performed. There were two variants of operation. In the first variant, invaginative valve from submucous-mucous membrane of the esophagus and the stomach was formed in the zone of gastroesophageal anastomosis. In the second variant, for prevention of anastomotic sutures eruption late after surgery and prophylaxis of valve expanding, duplication of muscular membrane on esophagus was formed. 58 patients were operated by the first variant, 17--by second variant. Postoperative lethality was 2.7% (1 patient died after operation of the first variant, 1 patient--after the second variant). In early postoperative period the complications were revealed in 8 (10.7%) patients. There was no insufficiency of anastomotic sutures. Mean hospital stay after operation was 14.7 +/- 3.5 bed days. Reflux-esophagitis of severe degree in remote period after operation was seen in 1 (1.3%) patient, catarral--in 4 (5.3%), all the patients were operated by the first variant. PMID- 11490486 TI - [Surgical prevention of postresection reflux-gastritis]. AB - Results of treatment of 331 patients operated in surgical clinic of Karaganda Medical Academy from 1990 to 1998 for complicated form of gastric and duodenal ulcers are presented. For surgical prophylaxis of postresective reflux-gastritis and to reduce the risk of sutures insufficiency two original methods of creation of transverse terminolateral gastroduodenoanastomosis with a reflux mechanism were proposed. These methods were used in 129 (39%) patients. Biochemical, endoscopic, X-ray, ultrasonic, morphological examinations demonstrated advantages of the proposed methods of stomach resection which promote portion-tardive type of evacuation with lower rate of postresective pathological states. It is concluded that these operations may be used as methods of choice in surgical treatment of ulcer. PMID- 11490487 TI - [Valvular pyloroplasty in the treatment of pyloroduodenal ulcers]. AB - Original method of treatment of complicated pyloroduodenal ulcers is proposed. The results of different organ-saving operations in complicated pyloroduodenal ulcers were analyzed in 136 patients aged from 15 to 89 years. Evaluation of the results was based on the data obtained during study of secretory and motor evacuatory function of the stomach, state of gastric and duodenal mucous membrane, degree of duodenogastric reflux. Gastroscopy in remote period after organ-saving operations enabled visual evaluation of state of anew formed pyloric canal. Results of treatment of complicated pyloroduodenal ulcers by different methods of pyloroplasty demonstrate the advantages of valvular pyloroplasty. PMID- 11490489 TI - [Portal embolization (an experimental-morphological study)]. AB - Results of experimental-morphological study of influence of unilobar portal embolization by biologic occlusive material RABROM on the liver of 6 laboratory animals are presented. It is shown that portal venous embolization leads to focal necrosis of parenchyma of embolized hepatic lobe, it atrophy and formation of portal cirrhosis. In non-embolized hepatic lobe the distinct signs of increased regeneration and hypertrophy of hepatocytes were revealed. RABROM didn't lead to damage and inflammatory changes of vascular wall that testifies to it biologic inertia. It is recommended to use the method of portal venous embolization for preparation of patients with low functional hepatic reserve for extensive resections. PMID- 11490488 TI - [Characteristics of immunologic disorders in patients with acute destructive pancreatitis]. AB - Examination of immunological status in patients with acute destructive pancreatitis with uncomplicated (14 patients) and complicated (infectious inflammatory processes--18 patients) postoperative period illustrated depression of T- and B-links of immunity, reduction of absolute and relative number of TFU- and TFC-lymphocytes. All the patients demonstrated reliable elevation of phagocytic rate, phagocytic index and number of circulating immune complexes. These changes were more significant in patients with complicated postoperative period. Level of lactoferrin in patients with complicated period was by 10% higher than in patients without complications. Significant elevation of tumor necrosis factor Ia in blood was registered in patients of both groups. During all the periods of examination the level of interleukin-8 was higher in patients with complicated postoperative period than in the patients with favorable postoperative period. This interleukin-8 is a reliable marker of postoperative complications in acute destructive pancreatitis. PMID- 11490490 TI - [Transurethral electrosurgery in the treatment of benign prostatic hyperplasia]. AB - The article is devoted to topical problem of modern urology, one of the lines of surgical treatment of benign prostatic hyperplasia (BPH)--transurethral electrosurgery. Physical principles which are the basis of transurethral electrosurgical interventions and influence of transurethral electrosurgery on prostatic tissue are described. Methods of necessary preoperative examination of patients with BPH are presented. The most often used transurethral electrosurgical technique for destruction of hyperplastic prostatic tissue is described. The results of 5-year use of combination of transurethral electroresection and electrovaporisation in the treatment of BPH and its complications are presented. Ways of prophylaxis of possible complications are proposed. PMID- 11490491 TI - [Long-term results of the surgical treatment of patients with multinodular euthyroid goiter]. AB - Long-term results of treatment of 280 patients operated for multinodal euthyroid goiter (MNEG) in surgical department of Endocrinology Research Center RAMS from 1983 to 1997 were studied. Hormonal examination of peripheral blood, ultrasonic examination of thyroid gland (TG) and zones of regional lymph outflow, thin needle punch biopsy were used as special methods of investigation. Correlation between recurrence and histological form was studied. In cystic-colloid goiter with adenomatosis the recurrence of MNEG was revealed in 15 (34.1%) cases. In primary multiple TG adenomas of different types the number of recurrences was smallest--1 (3.3%) cases. There were no recurrences in cancer of TG in combination with cystic-colloid goiter or adenomas of TG. Recurrence of cystic colloid goiter was revealed in 5 (5.7%) cases, different types of adenomas--in 5 (5.7%) cases. The smallest number of recurrences of MNEG was seen after maximum subtotal resection of TG--in 4 patients (7.7% of the patients with this volume of operation). In all the 12 cases of partial resection of TG the recurrence of different morphological forms of MNEG was revealed. Influence of thyroid therapy on remote recurrence rate in different form of MNEG was also studied. In cystic colloid goiter (CCG) of different proliferation degree with adenomatosis the recurrence rate was not significantly different irrespective of hormonal therapy- 29.4%, 37.5% and 36.4%, respectively, in adequate therapy, inadequate therapy and its absence, that testifies to small influence of thyroid therapy on reduction of recurrence rate in this group of patients. The highest recurrence rate in CCG and CCG with adenomas of TG was revealed in the group without thyroid therapy--30.8% and 35.7%, respectively. PMID- 11490492 TI - [Use of absorbable polymers for plastic repair of postoperative ventral hernias]. AB - In surgical clinic of Vladivostok State Medical University 48 patients were operated for postoperative ventral hernias with use of absorbable polymers. In group 1 (42 patients) the explant "Carbonicus-I" was used (period of absorption-2 years). 6 patients of group 2 underwent plastic repair with suture-glue duplication of aponeurosis of abdominal white line. The analysis of the results demonstrated that explants "Carbonicus-I" and tissue glue "MK-7M" can be used for plasty of hernial ring on anterior abdominal wall in large ventral hernias. Polymers with long period of absorption (over 2 years) form dense scar which prevents recurrence of hernia. At the same time polymers with short period of absorption (4-6 months) reduce possibility of early reaction of explant rejection. PMID- 11490493 TI - [Ambulatory surgical treatment of inguinal hernia]. AB - Results of ambulatory treatment of 4000 patients with inguinal hernias are presented. Principles of treatment with use of different methods and techniques are developed. Indications and contraindications for ambulatory treatment of patients with inguinal hernias are formulated. Results of treatment of 159 patients by original author's method of hernioplasty are presented. The authors consider ambulatory inguinal herniology as independent line in surgical practice. Foundation of ambulatory herniological centers are proposed. PMID- 11490494 TI - [State of the immune system in patients with penetrating thoracic and abdominal injuries complicated by massive hemorrhage]. AB - Results of immune status examination on day 1-2 and 4-8 after operation in 217 patients with penetrating gunshot and stab-incised wounds of thorax and abdomen were analyzed. Correlation between immunogram and volume of acute hemorrhage was studied. Types of immunograms characteristic of good and complicated postoperative period were determined, their frequency was estimated, that permits to perform substantiated and timely special correction for acceleration of immunorehabilitation and prevention of infectious complications. PMID- 11490495 TI - [Use of ozone therapy and hydro-pressure technologies in complex intensive therapy of surgical sepsis]. AB - Results of treatment of 214 patients with severe sepsis and septic shock were analyzed. 125 patients treated with various methods of ozonotherapy and hydropressive sanation of infectious foci formed the study group. Control group consisted of 89 patients treated according to generally accepted principles. Comparative analysis of treatment efficacy was carried out with numerous laboratory and instrumental study methods. It is shown that ozonotherapy and hydropressive technologies reduced a lethality from 39.2% in the control group to 25.6% in the study group. PMID- 11490496 TI - [Rapid method of bile detection in the abdominal cavity in closed abdominal trauma]. PMID- 11490497 TI - [Method of suturing duodenal stump in giant penetrating duodenal ulcers]. PMID- 11490498 TI - [Risk and prevention of venous thromboembolic complications in surgery]. PMID- 11490499 TI - [Beta blockers in therapy of hypertension. For obese patients only the second choice? (interview by Dr. Beate Schumacher)]. PMID- 11490500 TI - [Munchausen in general practice. Experiences with factitious disorder patients]. PMID- 11490501 TI - [Patient with factitious disorder. Searching for motives instead of exposure]. PMID- 11490502 TI - [Diagnosis of space-occupying lesions of the liver. What is the value of diagnostic imaging?]. AB - Ultrasound in B-mode or tissue harmonic imaging is the procedure of first choice for investigating the liver. If US is employed to screen for metastatic disease, B-mode contrast harmonic imaging should be used in addition to conventional ultrasound. For the diagnosis "space-consuming liver lesion", the echogenicity and clinical aspects determine the further diagnostic procedure. Echogenic liver lesions in patients with known malignant disease, and echo-poor or "echo-complex" lesions always indicate a need for further spiral CT or MRT investigations. In tumour patients, spiral CT offers the advantage of enabling simultaneous evaluation of the parenchyma, and abdominal staging. Primary SPIO-amplified MRT is indicated whenever the primary interest is the detection of space-consuming liver lesions. In the case of echogenic lesions in the absence of underlying malignant disease, US follow-up suffices. Here, differential diagnostic information can be gained from the vascular distribution patterns of a lesion obtained with color-coded duplex US. Hepatic lesions that, after exhausting all diagnostic imaging procedures remain suspicious, require biopsy. PMID- 11490503 TI - [Viruses, bacteria, drugs, allergies. What is behind exanthemas?]. PMID- 11490504 TI - [Every 8th patient suffers from onychomycosis. Eliminating the fungus! (interview by Petra Eiden)]. PMID- 11490505 TI - [Emergencies in general practice, 12. Hypoglycemic crisis]. PMID- 11490506 TI - [Anemias in general practice. Do not simply prescribe iron!]. PMID- 11490507 TI - [Secondary forms of hypertension, 1: Renal artery stenosis. How to recognize, how to treat?]. PMID- 11490508 TI - [Acute therapy of heart failure with nitroglycerin. Rapid help in routine practice]. PMID- 11490509 TI - [Alpha-glucosidase inhibitor in type 2 diabetic patients. Metabolism and body weight are favorably modified]. PMID- 11490510 TI - [Myeloproliferative diseases. Sometimes waiting it out is the best therapy]. PMID- 11490511 TI - The diagnosis of Sjogren's syndrome: definition and validation of classification criteria for this disorder. AB - Classification criteria currently exist for most of the rheumatic diseases. Different criteria sets have also been proposed in the past for Sjogren's syndrome (SS), but none of these has been validated and no single one is in general use by the entire scientific community. Between 1988 and 1996 the European Study Group for the Classification Criteria for SS performed a multicenter study in which a new classification criteria set for this disease was defined and then validated. This European set of classification criteria for SS is rapidly becoming the most widely accepted procedure to classify patients with the primary and secondary variants of the syndrome. PMID- 11490512 TI - Immunopathology of Sjogren's syndrome. AB - Sjogren's syndrome is a chronic autoimmune disorder characterized by mononuclear cell infiltration around epithelial cells of exocrine glands. In recent years, several studies have tried to elucidate the components of the immunopathologic interaction in Sjogren's syndrome as well as the function of these components. The majority of the mononuclear infiltrating cells are CD4 positive T lymphocytes (60-70%) whereas B cells constitute one fourth of the infiltrating cells. Macrophages and natural killer cells are poorly represented in the lesion. Epithelial cells of minor salivary glands of patients with Sjogren's syndrome express several cytokines (IL-1 beta, IL-6, NO), protooncogenes (c-myc), autoantigens (Ro, La, Fodrin) and costimulatory molecules (B71, B72). The characteristic destruction of epithelial cells of Sjogren's syndrome patients is probably due to activation of several apoptotic pathways since epithelial cells express different apoptosis related molecules such as Fas, FasL, Bax, while mononuclear cells express Perforin and Granzymes. Finally epithelial cells seem to exert a regenerative effort since they express trefoil proteins (pS2). The above mentioned properties give epithelial cells the leading role in the pathophysiology of the syndrome but the exact causative agent which drives the immune system towards an autoimmune reaction still remains obscure. PMID- 11490513 TI - The role of apoptosis in Sjogren's syndrome. AB - It has been suggested that defects in the modulation of programmed cell death/apoptosis might lead to autoimmune disease, such as Sjogren's syndrome (SS) and systemic lupus erythematosus. In this review some basic information on apoptosis is introduced together with three aspects on apoptosis in relation to SS: i) defective apoptosis could lead to lymphoid cell accumulation and chronic inflammation in exocrine glands; ii) increased apoptosis of epithelial cells might explain the loss of secreting epithelium; and iii) orderly destruction of cellular components might induce autoantibody production. Altogether, the idea that defects in the apoptotic process could be of importance for explaining autoimmune diseases, makes research on the different factors in this pathway valuable for achieving a better understanding of the etiopathogenesis of SS. PMID- 11490514 TI - Viral studies in rheumatic disease. AB - Rheumatic disease has long been thought to represent an interaction between environmental agents on a background of genetic susceptibility. In this review herpesviruses and retroviruses are considered as possible aetiological agents in autoimmune disease with a particular emphasis on Sjogren's syndrome. A possible role for cytomegalovirus, Epstein-Barr virus (EBV), human herpesvirus-6 (HHV-6) and human herpesvirus-8 (HHV-8) is reviewed. We conclude that there is no compelling evidence for the involvement of any of these herpesviruses. Retroviruses, however, are attracting increasing interest. In Man, both Human immunodeficiency virus (HIV) and human T lymphotropic virus type I (HTLV-I) infections cause autoimmune phenomena, including Sjogren's syndrome and arthritis in a minority of infected individuals. Similar reactions to retroviral infection are also seen in animal models. A possible role for the newly described human retrovirus-5 (HRV-5) is discussed, though current evidence does not support a role in Sjogren's syndrome. Other autoimmune diseases are under investigation. PMID- 11490515 TI - Antibodies to Ro and La. AB - Precipitating antibodies to Ro and La occur in a subset of patients with systemic lupus erythematosus (SLE) constituting just under 50 and 20% respectively of the total spectrum. These precipitating autoantibodies are even more prevalent in primary Sjogren's syndrome (SS) occurring in 60-80% (anti-Ro) and 40-60% (anti La) of that disease. Patients with an overlap of SLE and SS virtually all possess both anti-Ro and anti-La. These autoantibodies appear years before the appearance of clinical disease as evidenced by the behavior of women who possess them and herald this presence by having children born with the manifestations of neonatal lupus, principally a characteristic lupus dermatitis or complete congenital heart block. The close association of several clinical manifestations involving the skin, lung, and blood elements suggests a pathogenic role for these autoantibodies in disease expression. The elucidation of these relationships should greatly improve our understanding of the etiopathogenesis of both SLE and SS. PMID- 11490516 TI - Sjogren's syndrome: an historical perspective. PMID- 11490518 TI - Immunogenetics of Sjogren's syndrome. AB - The etiology for SS remains unknown where multifactorial influences contribute to the pathogenesis of subsequent development of the disease. The genetic influence is also multifactorial and the data suggests a familial component indicative of autosomal-dominant genes as well as genetic contributions associated with class-I and class-II HLA alleles. Various auto-antibodies found with increased frequency in SS (anti-Ro and anti-La) were shown to be associated with HLA class-II alleles at the DQA1 and DQB1 loci that were also found to have in common specific amino acid residues (10). Ethnic groups have also been studied and show varying HLA associations with primary SS. Multiple ethnic groups, however, share a DQ allele supporting the idea that the majority of SS patients carry a common allele which may predispose to primary SS. In some cases, the HLA-DR antigen may be induced and cause to appear on epithelial cells where they present antigen to CD 4+ T cells, which then go on to aid in the destruction of salivary gland epithelial cells specifically. The further elucidation of disease associations as well as possible immunogenetic pathogenic mechanisms may help to explain the causes for the development of various autoimmune diseases such as Sjogren's Syndrome. This may eventually result in the ability to immunomodulate these abnormal immune responses. In so doing, an approach to treatment by genetic engineering may also be possible once a further understanding of the genetic influences and mechanisms in the causation of autoimmune disease is further elucidated. PMID- 11490517 TI - Changes in glycosylation of immunoglobulins in primary Sjogren's syndrome. AB - Serum IgG and IgA are glycoprotein and significant glycoform abnormalities have been established in primary Sjogren's syndrome. The proportion of asialylated IgG is abnormally high in the patients, whereas IgA1 and IgA2 appear to be over sialylated. This peculiarity might explain the defective binding of IgA to asialoglycoreceptors. Furthermore, the activity of alpha 2,6 sialyl transferase is higher in the IgA-producing B cells from the patients than in the controls, whereas the alpha 2,3 sialyl transferase operates in the former cells but not in the latter. The mechanism of this enzyme dysregulation warrants elucidation. PMID- 11490519 TI - Therapy of Sjogren's syndrome. New aspects and future directions. AB - Therapy of Sjogren's syndrome remains empirical and symptomatic. The main goals are to treat the disease related features, especially sicca manifestations, since the immunosuppressive therapy has not given promising results. For the treatment of keratoconjunctivitis sicca: local stimulators of tear secretion, protective bicarbonate buffered solutions, replacement therapy or supportive operative procedures should be tried. For oral manifestations: stimulators of salivary secretion such as pilocarpine, or agents changing the consistency of saliva such as bromhexine orally should be tried. Saliva substitutes have a transient effect. Frequent ingestion of sugar free liquids may help. Oral hygiene is important to avoid oral candidiasis and dental caries. Treatment of parotid gland swelling is not necessary. Pulmonary manifestations include pulmonary infiltrates in perialveolar areas, nodular or cavitary lesions which may represent lymphoma. Hilar adenopathy, solid or cavitary lesions should be biopsied. In case of vasculitis prednisolone 1 mg/kg/day with progressive tapering should be tried. Renal involvement is manifested mainly as interstitial disease. Administration of NaHCO3 or sodium citrate is important to prevent acidosis and nephrocalcinosis. Vasculitis, when it is of the leukoclasic form, does not need therapy; when it is manifested with severe major organ involvement corticosteroids and/or cytotoxic therapy should be tried. Lymphoma is treated as in the patients without Sjogren's in close collaboration with the oncology department. PMID- 11490520 TI - Epidemiology of Sjogren's syndrome, especially its primary form. AB - During the last decades different sets of criteria have been used for defining primary Sjogren's syndrome. Using these criteria prevalence estimates have varied between 0.04%-4.8%. This hundred-fold distribution in estimated frequency is partly due to differences in age groups studied, classification criteria used and methods used for objective evaluation of lachrymal and salivary gland hypofunction. Another problem which makes comparison between studies difficult is that workshop recommendations for the various objective tests are very often not strictly followed. Population based studies tend to identify cases with less severe disease and higher prevalence rates. Furthermore, there seems to be little correlation between dryness symptoms reported by subjects and objective hypofunction of lachrymal and/or salivary glands. Possible explanations to these findings include selection bias towards more severe cases in clinical (university) studies and a low predictive value for some criteria in population studies. PMID- 11490521 TI - [Cutaneous manifestations of activated protein C resistance]. PMID- 11490522 TI - [Cardiovascular manifestations of pseudoxanthoma elasticum. Apropos of 2 cases]. PMID- 11490523 TI - [Retroperitoneal fibrosis associated with Takayasu arteritis]. PMID- 11490524 TI - [Bronchocentric granulomatosis syndrome and actinomycosis caused by Actinomyces meyeri]. PMID- 11490525 TI - [Drug-induced fever caused by hydroxyurea. Apropos of 3 new cases]. AB - Drug-induced fever is a frequent (3-5% of all adverse effects) but under recognized adverse effect of several drugs. Hydroxyurea, an antimetabolite cytostatic agent, has rarely been involved in the occurrence of fever. We report three additional cases of hydroxyurea-induced fever including one case with pulmonary involvement (acute alveolitis). In these cases, the role of hydroxyurea was strongly suggested by the delay to onset of symptoms (16-36 days), the disappearance of fever within a few hours after drug withdrawal, the recurrence of fever shortly after rechallenge in two patients, and the absence of any other obvious cause. PMID- 11490526 TI - [Cerebral toxoplasmosis in HIV infected patients intolerant of cotrimoxazole]. AB - Using results of a multicentric randomized prospective trial of primary prophylaxis of Pneumocystis carinii pneumonia in HIV-infected patients which compared sulfamethoxazole-trimethoprim and pentamidine isethionate, the risk to develop cerebral toxoplasmosis was analyzed in the two assigned groups and in the groups of patients who stopped sulfamethoxazole-trimethoprim prophylaxis. The risk to develop cerebral toxoplasmosis appeared significantly higher in the group of patients who stopped sulfamethoxazole-trimethoprim consecutively to cutaneous hypersensitivity. PMID- 11490527 TI - Renal ostodystrophy during the developing stage of maintenance dialysis in Transylvania. Early development of periarticular calcifications and beta 2 microglobulin amyloidosis in spite of a relatively good prevention of secondary hyperparathyroidism. AB - BACKGROUND: Dialysis facilities have been introduced only recently in Transylvania with many limitations, in particular a standard high calcium dialysate, Al(OH)3 as phosphate binder and pharmacological doses of native vitamin D2, but neither CaCO3 nor 1 alpha hydroxylated vitamin D. Rheumatological complaints and metastatic calcifications were frequent, leading to suspect either overt hyperparathyroidism, adynamic bone disease or beta 2 microglobulin amyloidosis. AIMS OF THE STUDY: Evaluate the prevalence of radiological osteitis fibrosa, amyloid osteoarthropathy and periarticular calcification and their link with PTH secretion, phophocalcic disorders, acidosis, bone turn over, aluminum and beta 2 microglobulin accumulation in the dialysis population of Sibiu (Transylvania). METHODS: The clinical and radiological rheumatological data of the 49 uremic patients dialyzed in Sibiu since 1990 were reviewed as well as the monthly routine monitoring of their plasma phosphocalcic parameters. Furthermore in July 1994, 36 of them had an X rays of the hands for evaluation of subperiosteal resorption of the phalanges, periarticular calcifications and carpal cysts as well as a determination of plasma concentrations of intact PTH (normal range: 10-55; optimal range: 100-200 pg/ml), osteocalcin, bone alkaline phosphatase, aluminum and 25 OH vitamin D. RESULTS: The prevalence of subperiostal resorption of the phalanges was 8% and that of severe biological hyperparathyroidism (PTH > 400 pg/ml) 22%, whereas that of a relative hypoparathyroidism (PTH < 100 pg/ml) was 31%. Mean plasma concentrations of calcium was 2.07 +/- 0.15; of phosphate 2.50 +/- 0.35; of bicarbonate 15 +/- 2.0 mmol/l, of 25 OHD 30 +/- 20 ng/ml, of aluminum 1.1 +/- 0.5 mumol/l. Plasma PTH concentrations were negatively correlated to dialysis duration, and to plasma concentrations of aluminum, calcium and 25 OH vitamin D but not to those of phosphate and bicarbonate. Multivariate analysis showed however that only duration of dialysis and plasma aluminum concentration were independently and negatively correlated to plasma PTH concentrations. The prevalence of periarticular calcifications (26%) and of carpal cysts suggestive of beta 2 microglobulin amyloidosis (10%) were relatively high considering the young age of the population (42 years) and the short duration of dialysis (2.6 years). Patients with calcifications comparatively to those without calcifications were older, had longer duration on dialysis, higher prevalence of carpal cysts and higher plasma beta 2 microglobulin concentrations, lower plasma PTH (98 versus 313 pg/ml) and higher plasma aluminum concentration (1.3 versus 0.8 mumol/l). Patients with carpal cysts comparatively to those without cyst were older, had a longer duration on dialysis and a higher prevalence of periarticular calcifications. CONCLUSIONS: a) In spite of no use of 1 alpha hydroxylated vitamin D derivatives, and poor control of hyperphosphatemia and acidosis, hyperparathyroidism declined with duration of dialysis due to the use of a high dialysate calcium concentration, Al(OH)3 as sole phosphate binder and high supplement of native vitamin D. b) Considering the relative young age and short duration on dialysis, the prevalence of periarticular calcifications and carpal cysts were high. c) Calcifications were possibly favored by relative hypoparathyroidism and moderate aluminum overload. d) The association of periarticular calcifications and subchondrial cysts suggest a causal relationship. PMID- 11490528 TI - Idiopathic osteoporosis: a heterogeneous entity. AB - DEFINITION: Idiopathic osteoporosis refers to the development of osteopenia and fractures with minimal or no trauma in otherwise young, healthy individuals who are not postmenopausal or have other, identifiable secondary causes of osteoporosis. EPIDEMIOLOGY: It is a relatively rare disorder, with an incidence of 0.4 cases per 100,000 person-years. It appears to affect both sexes equally and results primarily in the development of trabecular bone fractures such as vertebral compression fractures and Colles' fractures, although hip fractures are also seen. PATHOPHYSIOLOGY: The disease may be temporally related to pregnancy and/or lactation in some patients, although it is unclear whether pregnancy plays a pathophysiological role or, more likely, simply leads to the clinical presentation of the disease in individuals who are already affected. Various pathophysiological abnormalities have been described in these patients, including hypercalciuria, abnormalities in vitamin D metabolism, and in the production of insulin-like growth factor I and interleukin 1. Findings on bone biopsy have been variable, with some patients having evidence of a defect in osteoblast function, whereas others having evidence for increased bone resorption. TREATMENT: No specific therapy has been proven to be effective in these patients. However, an individualized approach based on an assessment of bone turnover may be reasonable and may decrease the bone loss and subsequent fracture risk. PMID- 11490529 TI - [Leukemic risk in thrombocythemia]. AB - OBJECTIVES: To define pathogenesis, incidence, clinical presentation and prognosis of leukemic transformation (LT) in patients with essential thrombocytemia (ET); to compare the incidence of LT in previously treated versus untreated patients; to search for risk factors of LT, with special reference to the iatrogenic risk of myelosuppressive drugs. DATA SOURCES AND EXTRACTION: Most significant French and English-language papers published between 1981 and 1997 dealing with either ET, LT, clonality, cytogenetics, alkylating agents or hydroxyurea were reviewed. Patient characteristics, clinical presentation, bone marrow and cytogenetic findings, and survival data were extracted from each case report and review, and possible risk factors were analyzed. DATA SYNTHESIS: Despite the existence of clonal hematopoiesis in up to 60-80% of patients with ET, suggesting an intrinsic potential for subsequent leukemia, spontaneous LT is poorly documented and the common presumption that TL occurs in excess in treated patients with ET is still questionable. The incidence of LT, probably underestimated in the past, varies from 0.7% to 5.3% in retrospective series, and from 8% to 12% in prospective studies. Recurrent cytogenetic abnormalities are significantly more frequent and complex in marrow karyotypes performed at the time of blastic crisis, as compared with those performed at diagnosis. A review of 62 reports of LT highlights some important characteristics: a) LT can occur anytime during the chronic phase of ET, especially within the first eight years, b) all subtypes of leukemia are described, notably unclassifiable cases and megakary oblastic acute leukemias, c) bone marrow fibrosis is frequent, as is a progressive onset of a myelodysplastic syndrome preceeding the blastic crisis, d) the prognosis of LT is very poor, two-third of all patients die within 6 months. Apart from the onset of a myelodysplastic syndrome, which carries a poor outcome, there is no well-established risk factor of LT. The leukemogenic risk of myelosuppressive therapies, clearly demonstrated for years in polycythemia vera, cannot be directly ascertained in patients with ET, but is suggested by various indirect observations. This putative risk of inducing secondary leukemia affects not only alkylating agents but also hydroxyurea. CONCLUSION: LT is a highly serious and largely unpredictable complication of ET. The current debate on the specific risk of myelosuppressive treatments deserves large-scale prospective randomized studies. Until this crucial point is elucidated, any cytotoxic therapy should be reserved to elderly and/or symptomatic patients and to those having other vascular risk factors. PMID- 11490530 TI - [Treatment of polycythemia. I--Using radiophosphorus with or without treatment in 483 patients over 65 years of age]. AB - AIMS: To compare by a prospective study in high risk polycythemia vera (PV) patients 33P alone and 32P followed by low-dose hydroxyurea (HU) maintenance therapy. Toxicity, efficiency, and leukemogenic potential were studied. PATIENTS: 483 patients with a documented PV, aged more than 65 years at diagnosis, were included between 1980 and 1996 in a prospective study comparing 32P alone and 32P followed by low-dose HU maintenance therapy. Blood cell counts were performed every two months and a clinical evaluation by a specialist was obtained every four or six months. RESULTS: Treatments were well tolerated, but chronic leg ulcers were observed in the maintenance therapy arm. The risk of leukemia was about 15% at the 15th year in the group of patients treated by 32P alone, but reached 30% in the group receiving maintenance therapy. In both arms, there was no significant correlation between occurrence of leukemia and the total dose of 32P. There was a correlation between the leukemic risk and disease severity, estimated on the frequency of relapse. Cancer occurrence was slightly higher than expected in the maintenance arm. HU treatment did not protect against progression to myelofibrosis, probably due to the lack of maintenance of an efficient myeloid or megakaryocytic suppression. Median life-span was slightly shorter in the group receiving HU maintenance. In all cases, life-span was only one year lower than that observed in the reference population. CONCLUSION: For all these reasons, we suggest the us of 32P alone in elderly patients; complementary chemotherapy should only be prescribed in the cases with short-term relapse, and late resistance to 32P. PMID- 11490531 TI - [Treatment of polycythemia. II.--Comparison of hydroxyurea with pipobroman in 294 patients less than 65 years of age]. AB - AIMS: To compare by a prospective study in low risk polycythemia vera (PV) patients alone two drugs: hydroxyurea and pipobroman. Toxicity, efficiency, and leukemogenic potential were studied. PATIENTS: 294 patients with a documented PV, aged less than 65 years, have been included since 1980 in a prospective study comparing hydroxyurea and pipobroman. Blood cell counts were performed every two months and a clinical evaluation by a specialist was obtained every four or six months. RESULTS: Hematologic toxicity of both drugs was higher than expected, requiring strict surveillance. These drugs were tolerated in some (gastric pain and diarrhea on pipobroman, buccal aphtosis and chronic leg ulcers on hydroxyurea), leading to a change of arm in 10% of the cases. Hydroxyurea did not control the megakaryocitic hyperplasia in 40% of the cases, which probably explains a high rate of progression to myelofibrosis with myeloid metaplasia in this arm. Both drugs were leukemogenic with an actuarial risk of about 15% at the 15th year, not significantly lower than that observed in the 32P treated patients. A significant risk of cutaneous malignancy was observed in the hydroxyurea arm. The mean expectancy of life cannot yet be accurately evaluated, but seems significantly lower than that of the reference population. CONCLUSION: The treatment of PV by hydroxyurea or pipobroman has to account for these results less optimistic than those traditionally well-known to hematologists and internists. PMID- 11490532 TI - [Chronic pseudo-hypertrophic myopathy in sarcoidosis. Apropos of 3 cases]. AB - We report three cases of pseudohypertrophic chronic myopathy in patients with sarcoidosis. The patients were aged 41-50 years and were of African or mixed ethnic origin. All three patients had multiorgan sarcoidosis recently diagnosed by the presence of hiliary and/or mediastinal node enlargement with or without reticulomicronodulation in the lung fields. Hypertrophy of the proximal portion of the limbs dominated the clinical presentation. In 2 cases, the myopathy was preceded by myalgia and in 2 cases associated with muscle nodules. The muscle enzymes were elevated in 2 cases. The importance of tuberculoid granulomas and the discretion of the myopathy characterised the histological findings in all 3 cases. Cortisone therapy was given early and long-term treatment led to favorable course in 2 cases, probably due to the importance of the granulomatous muscle mass. In the third case, peripheral neuropathy developed. PMID- 11490534 TI - [The spondylarthropathy concept. Prognosis and treatment]. PMID- 11490533 TI - [Still's disease in the adult]. PMID- 11490535 TI - HLA-B27 transgenic rats model. AB - To investigate the role of HLA-B27 in the pathogenesis of spondylarthropathies, rats transgenic for HLA-B27 and human beta 2-microglobulin were produced. Several lines of B27 transgenic rats spontaneously develop a multisystem inflammatory disorder reminiscent of human spondylarthropathies, with gut, joint, skin and male genital lesions. The role of HLA-B27 in this model was studied and it was found that a high expression level of the transgene in cells of hematopoietic origin was critical to induce inflammatory manifestations. Furthermore, a combined implication of CD4+ T cells and antigen presenting cells in this model is suspected, based on passive transfer studies, although a specific influence of HLA-B27 upon T cell education in the thymus is unlikely to be important. Beside the immune system, bacterial flora exerts an important influence in this model, as a triggering agent of gut and joint inflammation. Finally, endogenous rat genetic factors are suspected to be involved in this model. PMID- 11490536 TI - [Subtypes of the HLA-B27 molecule and association with spondylarthropathies]. AB - HLA-B27 subtypes differ in their ethnic distribution and in their susceptibility to spondylarthropathies (SA). B*2705 and B*2702 are the most frequent disease associated subtypes in Caucasians as well as B*2704 and B*2707 in Asia while B*2706 in Asia and B*2709 in Sardinia have been reported not to be associated to SA. Differences in antigenic peptide presentation could underlie such behavior. Several studies suggested that a Tyr C-terminal peptide anchor could be found preferentially in disease-associated subtypes and could be therefore one of the criteria in the search of putative arthritogenic peptide(s). We analyzed by HPLC and Edman sequencing peptides eluted from immunopurified HLA-B27 molecules expressed on B-lymphoblastoid cell lines or C1R transfectans of human origin. We focused our work on B*2707, associated with SA in the same geographical area where B*2706 is not. We found the same preference for Leu at the C-terminus in the peptides bound by both subtypes without any significant signal for Tyr. In the same experimental conditions a Tyr C-terminal anchor was found for B*2705, B*2702, B*2704, B*2703 and also for B*2701 and B*2708, 2 rare subtypes for which binding specificity was previously unknown. Comparison of the F-pocket aminoacid composition in these various subtypes showed a correlation between Asp at position 116 and Tyr at the peptide C-terminus. Asp116 is changed for Tyr in B*2706, B*2707 and His in B*2709, all subtypes allowing a Leu C-terminal anchor. Therefore a Tyr C-terminal anchor correlates with the HLA-B27 F-pocket composition rather than with susceptibility to SA. PMID- 11490537 TI - [Follow-up of the patient with spondyloarthropathy]. AB - Spondylarthropathies consist of several disorders: reactive arthritis, psoriatic arthritis, arthritis associated with inflammatory bowel disease, a subgroup of juvenile chronic arthritis, and ankylosing spondylitis. Their clinical presentation may consist in the following inter-related features: axial involvement, peripheral articular involvement, enthesiopathic lesions, extra articular disease. The monitoring of these diseases is related more to their clinical presentation than to the precise diagnosis. Modalities for monitoring peripheral arthritis are similar to those of rheumatoid arthritis (essentially based on the number of tender and swollen joints). The modalities of the monitoring of extra-articular features (uveitis, psoriasis,...) are specific to these clinical features and can be categorized in 2 ways: the first one consists in considering the occurrence of the episodes (for example, number of acute anterior uveitis per year), the second one consists in the evaluation of the severity of the clinical features (for example the area of psoriatic skin lesions). Numerous tools have been proposed to evaluate the axial involvement of the disease. The international rheumatologic community (ILAR for International League Against Rheumatism) via specific task force (ASAS for Assessment in Ankylosing Spondylitis) tried to standardize the medical language (at least in clinical research studies) by giving recommendations to evaluate specific domains and within each domain specific tools. Currently, it is generally agreed that pain and functional impairment are the two main domains to consider. For each of these domains, different tools have been proposed (for example the ASFI: Ankylosing Spondylitis Functional Index and the BASFI: Bath Ankylosing Functional Index are both tools proposed to evaluate the functional impairment). The domain "range of motion" is probably one of the most important for long term outcome in clinical research studies and for facilitating the orthopedic indications in daily practice. Finally, radiological tools permitting the evaluation of the structural damage of the disease are available. They take into account the presence and/or the severity of hip and sacroiliac involvement and also the extent of the spinal syndesmophytes. Longitudinal studies are required to evaluate their clinical relevance. PMID- 11490538 TI - Spondylarthropathies in children. AB - Spondylarthropathies in children are still classified among juvenile chronic (rheumatoid arthritides, at least in their undifferentiated forms. However, the ESSG and Armor criteria are very satisfactory for identifying this group of patients. Work conducted by the ILAR Taskforce for Pediatric Rheumatology is in progress for a better identification of these patients. PMID- 11490539 TI - [Palinopsia associated with partial seizures secondary to right parietal hemorrhage]. AB - Palinopsia and motor partial seizures were associated in a 51-year-old man. The manifestation appeared six months after a right parietal hemorrhage. The relationship between the hallucinatory phenomena and epilepsy is discussed. PMID- 11490540 TI - [Pulmonary sarcoid-like granulomatosis associated with Hodgkin's disease and complicated by bleomycin-induced pulmonary nodules]. AB - We report the case of a man in whom multiple lung macronodules developed after chemotherapy containing bleomycin for Hodgkin's disease, which was itself shortly preceded by a pulmonary sarcoid-like granulomatosis. Biopsy of the nodules showed fibrotic and granulomatous lesions. The etiological diagnosis is discussed. PMID- 11490541 TI - [Pleuropneumopathy caused by bromocriptine in a patient with Parkinson disease. Review of the literature apropos of a new case]. AB - Bromocriptine, a derivate of ergot of rye, is employed in high doses for the treatment of Parkinson's disease, and may induct pleuropulmonary affections. We report the case of a 75-years-old patient, on bromocriptin for 5 years, who presented a progressive dyspnea, due to a pleural thickening, and moderate interstitial infiltrate, associated with an inflammatory syndrome. The outcome was favorable with 3 months of bromocriptin cessation. These complications are rarely described, probably underestimated, and justify a respiratory follow-up for these patients, who are difficult to stabilize in their neurologic treatment. PMID- 11490542 TI - [Emperor Maximin I: a case history of gigantism in acromegaly]. PMID- 11490543 TI - Alzheimer's disease. From the syndrome to the management of the patients and their family. PMID- 11490544 TI - Epidemiology of dementia. AB - With an unprecedented demographic change in the world population, the prevention and effective treatment of age-dependent diseased has become a compelling need. Epidemiology contributes in this way by estimating the incidence and prevalence of specific categories of morbidity and by identifying the risk or protective factors of dementia. An integrative analysis of 47 surveys across 17 countries has suggested approximate rates under 1% for dementia from any cause in persons aged 60 to 69 years, rising to about 39% in persons 90 to 95 years old. The prevalence doubles with every five years of age within that range, with few differences taking into account secular changes, age, gender, place of living... Risks factors of vascular dementia are well known. Worldwide epidemiological surveys allow to differentiate "confirmed risk factors" of Alzheimer's type dementia (age, family history, Apolipoprotein E epsilon 4 genotype and Down's syndrome), "possible risk factors" (head injury, aluminum, previous depression...) and "protective factors" (education, anti-inflammatory drugs and estrogen replacement...). PMID- 11490545 TI - Cellular vulnerability in brain aging and Alzheimer's disease. Clinical correlates and molecular background. AB - The neuropathological changes associated with normal brain aging and Alzheimer's disease involve specific cortical circuits. Extensive hippocampal alterations are correlated with age-associated memory impairment, while substantial neurofibrillary tangle formation in neocortical association areas of the temporal lobe is a prerequisite for the development of Alzheimer's disease. Several lines of evidence indicate that there is no correlation between senile plaque densities and the degree of dementia in this disorder. The cortical involvement in the ninth and tenth decades of life is different from that observed in younger patients in that parietal and cingulate areas are affected early in the course of Alzheimer's disease, and neocortical senile plaques densities are strongly correlated with the severity of dementia. Moreover, Alzheimer's disease pathology is characterized in these very old patients by high neurofibrillary tangle densities in the anterior CA1 field, but not in the entorhinal cortex and inferior temporal cortex. These patterns of lesion distribution are discussed in respect to the neurochemical, genetic and metabolic factors which may influence the neuronal vulnerability in Alzheimer's disease. PMID- 11490546 TI - [Early diagnosis of Alzheimer disease]. AB - On of the major challenges for public health in developed countries for the 21st century is that of aging, and in particular Alzheimer's disease. Progress has been made in the development of symptomatic treatment and prevention and early diagnosis of Alzheimer's disease is becoming mandatory. In the present state of our knowledge, no marker for early diagnosis has been described. However, the combined use of clinical observations (behavioral and neuropsychological features) and complementary information (brain morphology and functional imaging, biological data, neurophysiology explorations) could be helpful for early diagnosis of Alzheimer's disease well before the onset of dementia. PMID- 11490547 TI - Vascular dementia. Differential diagnosis and therapeutic issues. AB - Although it has long been felt that dementia may be due to atherosclerosis, the concept has recently evolved to include multiple pathophysiological mechanisms related to deficiencies in cerebral blood supply. Epidemiological data has identified hypertension and stroke as the most potent risk factors for the development of vascular dementia (VaD). New diagnostic criteria have been proposed and new neuroimaging techniques have led to a better detection of cerebral vascular pathology. However, the differential diagnosis between Alzheimer's disease and VaD, the two most common causes of dementia, remains clinically challenging. Therapeutic interventions for VaD are limited, nevertheless several lines of evidence suggest a strong potential for preventive treatment through the control of vascular risk factors. PMID- 11490548 TI - Dementia with Lewy bodies. AB - The presence of a high number of Lewy bodies--the morphological marker of Parkinson's disease--in the cerebral cortex of some cases of dementia has been frequently observed in association to Alzheimer type lesions (mainly senile plaques) and changes in the substantia nigra, that may be held responsible for the frequently associated symptoms of parkinsonism. The term "dementia with Lewy body" (DLB) has recently been suggested by a consensus conference and indicates that the pathogenetic mechanism of the dementia remains poorly understood. Marked fluctuations of alertness and of the cognitive performances, moderate parkinsonism and episodes of visual hallucinations may lead to suspect this diagnosis in cases of dementia. Unexplained falls, syncopes, delirium or alterations of consciousness may also be observed, and the patients may then be admitted in departments of internal medicine or geriatrics. The Lewy body is an intraneuronal spherical inclusion, present in Parkinson's disease. It is observed in the brainstem (substantia nigra, locus coeruleus, dorsal nucleus of the Xth nerve) and in the nucleus basalis of Meynert. The cortical Lewy bodies have a different aspect, but retain their antigenic characteristics: they are, in particular, stained by the antiubiquitin antibodies. Recently, they were found to be also labeled by antisynuclein antibodies. A mutation of the synuclein gene was recently identified in cases of familial Parkinson's disease. Clinically as well as pathologically, DLB may thus be difficult to distinguish from Alzheimer's disease on the one hand, and from Parkinson's disease, on the other. That diagnosis, however, is associated with a poor prognosis and should lead to specific therapeutic measures. PMID- 11490549 TI - Behavioral abnormalities in Alzheimer's disease. AB - Fundamental and therapeutic research into Alzheimer's disease (AD) initially focused on cognitive aspects. However, AD also frequently involves complex affective and behavioral disorders, which are currently grouped under the heading Behavioral and Psychological Signs and Symptoms of Dementia (BPSSD). BPSSD further handicap the patient in his or her daily activities and are a major source of problems for the family and other carers. Studies of these aspects must take into account various clinical aspects such as the development of reliable tools for assessment and diagnosis. BPSSD can only be understood on the basis of neuroanatomical and neurochemical findings, i.e. the link between the dysfunction of cortical-subcortical regions and networks and the onset of a particular behavioral disorder. Finally, therapeutic research must go beyond simple adaptation of an adult treatment protocol to affected elderly subjects by learning more about the different neurotransmitter systems in Alzheimer's disease. PMID- 11490550 TI - Prevention of Alzheimer's disease. AB - Delaying the emergence of Alzheimer's disease (AD) symptoms by 5 to 10 years would have a significant public health impact in the long term. Data generated from large scale epidemiological studies on risk as well as protective factors for AD are leading to treatment strategies that can be tested in transgenic animal models as well as human populations at higher genetic risk. PMID- 11490551 TI - Treatment of Alzheimer's disease. New developments. AB - Cholinesterase inhibitors (ChEI) represent the drug of choice for Alzheimer's disease treatment. They produce significant improvement on cognitive as well as non-cognitive function for a period up to one year during the first three years period following clinical onset. The magnitude of cognitive improvements is similar for different ChEI; however, differences are seen with regard to incidence and severity of side effects, incidence of drop-outs, optimal ChE inhibition, pharmacokinetics properties and mode of administration. PMID- 11490552 TI - The Ontario Hospital Reporting System: looking good, getting better. PMID- 11490553 TI - Building a national nursing agenda. A timely response for the sickest workers in the country. PMID- 11490554 TI - The First Ministers' Communique on Health: provincial health reform joined by federal health dollars. PMID- 11490555 TI - Operational review: the London Health Sciences Centre experience. PMID- 11490556 TI - The ministry perspective. PMID- 11490557 TI - Canada's specialty care system: the need for urgent reinvestment and rejuvenation. AB - Later this year, the Canadian Medical Association will publish a background discussion paper on Canada's specialty care delivery system entitled, "An Overview of Specialty Care in Canada: Issue Identification and Policy Challenges." This article draws on the information contained within this discussion paper. PMID- 11490559 TI - Physician recruitment: what hospitals need to know. PMID- 11490558 TI - The role of the physician executive in managing the healthcare value chain. PMID- 11490560 TI - A strategy for recruiting and retaining health information management professionals. AB - In the summer of 1998, the University Health Network (UHN) faced an acute personnel situation in which an exodus of skilled resources was taking place. The turnover of information management staff was increasing dramatically, and despite best efforts to recruit new employees, the vacancy rate remained at an unsustainable level. This case study reviews both the short- and long-term strategies that were employed to address these issues and to establish UHN as an employer of choice in health information management. Material in this article is based in part on papers presented at HIMSS 2000 and INFOCUS 2000 (Cameron, Claiter and Murphy 2000). PMID- 11490561 TI - Restructuring a national treasure: the new plan for England's National Health Service. PMID- 11490562 TI - Where is the patient in healthcare change? PMID- 11490564 TI - Strategic imperatives that will drive healthcare in a networked world. PMID- 11490565 TI - Wiring a new business model for healthcare procurement. PMID- 11490566 TI - The new healthcare accord: will it change public opinion? PMID- 11490567 TI - Nutritional modulation of the carcinogenesis process: targets and examples. PMID- 11490568 TI - Nutrition and breast cancer: epidemiology and mechanisms. PMID- 11490569 TI - The European prospective investigation into cancer and nutrition: perspectives for cancer prevention. PMID- 11490570 TI - Catabolism of skeletal muscle proteins and its reversal in cancer cachexia. PMID- 11490571 TI - New mediators in cancer cachexia. PMID- 11490572 TI - Animal models for nutrition in cancer. PMID- 11490573 TI - Bidirectional interplay of nutrition and chemotherapy. PMID- 11490574 TI - Nutrition in hematopoietic stem cell transplantation. PMID- 11490575 TI - Nutritional support of patients during radiotherapy. PMID- 11490576 TI - Epidemiology of nutrition and lung cancer. PMID- 11490577 TI - Nutrition in oncological surgery. PMID- 11490578 TI - Is immune nutrition the Holy Grail for cancer patients? PMID- 11490579 TI - Nutritional support in pediatric cancer patients. PMID- 11490580 TI - Workshop on what constitutes an optimal biomarker in cancer trials. PMID- 11490581 TI - Antioxidants and lung cancer prevention. PMID- 11490582 TI - Prostate cancer: epidemiology and prevention. PMID- 11490583 TI - Nutrition and colon cancer prevention. PMID- 11490584 TI - The mechanisms by which folate depletion enhances colorectal carcinogenesis: a unified scheme. PMID- 11490585 TI - Comprehensive geriatric assessment (CGA) and the MNA: an overview of CGA, nutritional assessment, and development of a shortened version of the MNA. PMID- 11490586 TI - The Mini Nutritional Assessment for preoperative nutritional evaluation: a study on 419 elderly surgical patients. PMID- 11490587 TI - The Mini Nutritional Assessment in clinical practice. PMID- 11490588 TI - The Mini Nutritional Assessment and body composition in healthy adults. PMID- 11490589 TI - MNA and nutritional intervention. PMID- 11490590 TI - MNA and cost of care. PMID- 11490591 TI - Animal model: metabolic and thermic responses to diet and environment (4 degrees C) in obesity during aging in the LA/Ntul//-cp rat. AB - The results of this study indicate that aging is associated with progressive declines in the normal dietary and metabolic responses to diet and environment in this animal model. The normal thermic and hormonal responses to diet and environment are thought to contribute to the fine regulation of energy balance by modulation of the efficiency of energy utilization or storage [25]. In the obese phenotype, the declines in the above variables may be further complicated by the presence of long-standing insulin resistance, and by perturbations in the normal metabolism and action of thyroid hormones, particularly T3. These may further contribute, individually or in combination with other factors, to an enhancement of energy storage and thereby contribute to maintaining the obese state. Moreover, the effects of progressive increases in obesity further exacerbate the decline in the economy of energy utilization and storage. Thus the impact of these physiologic changes of aging and obesity may impart significant alterations on the efficiency of energy metabolism, which in turn may contribute to some of the pathophysiologic changes associated with longevity, and could influence nutritional indices in affected individuals. As nations become progressively more industrialized, the incidence of overweight conditions--including obesity, NIDDM, and related metabolic disorders--has been shown to become increased and, along with those changes, the metabolic and pathophysiologic sequelae related to those disorders become more common [6-8]. A greater understanding of mechanisms of impaired energy metabolism and energy balance in aging may provide new insight into the nutritional factors that may contribute to obesity in aging, their modulation, and the emergence of a longer, healthier lifestyle. PMID- 11490592 TI - MNA and immunity: nutritional status and immunological markers in the elderly. PMID- 11490593 TI - The Mini Nutritional Assessment (MNA) for grading the nutritional state of elderly patients: presentation of the MNA, history and validation. PMID- 11490594 TI - Comparative nutrition evaluation with the Mini Nutritional Assessment and the Nutritional Risk Assessment Scale. PMID- 11490596 TI - The MNA score in people who have aged successfully. PMID- 11490595 TI - MNA and odor perception. PMID- 11490597 TI - Anorexia of aging, leptin, and the Mini Nutritional Assessment. AB - In this chapter we have reviewed the evidence for physiological anorexia of aging and stressed that its pathophysiology involves both central and peripheral mechanisms. Early satiation in the older person appears to involve signals predominantly arising in the stomach. The increased feeling of satiety in older persons is mainly related to changes in the central feeding drive, in particular a decrease in the opioid rewarding properties for fatty foods. Increased cytokines, secondary to inflammatory conditions which are common in old age, may further increase the anorexia seen in older persons. Leptin, the fat hormone, is an excellent indicator of fat mass in women, in whom leptin concentrations correlate with the MNA. In men, testosterone inhibits leptin, and the fall in testosterone with age results in an increase in leptin concentrations. In males the MNA is not related to leptin concentrations. Finally, we have examined the interrelation of two nutritional screening indices, MNA and SCALES. The two indices were well correlated and were both predictive of poor basic function. We conclude that the MNA is an excellent predictor of nutritional status. These findings suggest that malnutrition is a major predictor of frailty or the "failure to thrive" syndrome in older persons. Depression is a major cause of poor nutritional status in older persons. PMID- 11490598 TI - Associations among the Mini Nutritional Assessment instrument, dehydration, and functional status among older African Americans in St. Louis, Mo., USA. PMID- 11490599 TI - Mini Nutritional Assessment and Alzheimer patients. PMID- 11490600 TI - Mini Nutritional Assessment and cancer. PMID- 11490602 TI - Physicochemical considerations of protein utilization. PMID- 11490601 TI - Proteins, peptides and amino acids in enteral nutrition: overview and some research challenges. AB - We have selectively reviewed various aspects of protein and amino acid metabolism and nutrition which we feel are relevant and important in any comprehensive consideration of proteins, peptides, and amino acids in enteral nutrition. These, together with various clinical aspects of the problem that we have not addressed, are covered in more specific detail elsewhere in this volume. Our hope is that by the time this workshop is at an end a reasonable picture of the state of the art will emerge in terms of an understanding of the physiology and biochemistry of protein, peptide, and amino acids in enteral nutrition. Simultaneously, we will need to know, through careful clinical investigation, how this knowledge can best be exploited for optimizing the design of the protein/peptide/amino acid component of enteral nutrition formulations. In doing so it should be possible to achieve a more effective enteral use of proteins, peptides, and amino acids in the comprehensive clinical management of individuals under various pathophysiological states. PMID- 11490603 TI - The 'fast' and 'slow' protein concept. PMID- 11490604 TI - Postprandial protein utilization: implications for clinical nutrition. PMID- 11490605 TI - Catabolic states and immune dysfunction: relation to gastrointestinal feeding. PMID- 11490606 TI - Cysteine and glutathione in catabolic states. PMID- 11490607 TI - Conditionally indispensable amino acids (glutamine, cyst(e)ine, tyrosine, arginine, ornithine, taurine) in enteral feeding and the dipeptide concept. PMID- 11490609 TI - Role of enteral nutrition in the pathophysiology and treatment of pancreatitis and cystic fibrosis. PMID- 11490608 TI - Tolerance and utilization of enteral nitrogen. PMID- 11490610 TI - Nutrition in renal failure--the role of enteral feeding. PMID- 11490611 TI - Role of the gut in the amino acid economy of the host. PMID- 11490612 TI - Proteins, peptides and amino acids: which and when? PMID- 11490613 TI - Regulation of splanchnic protein synthesis by enteral feeding. PMID- 11490614 TI - Mechanisms of peptide and amino acid transport and their regulation. PMID- 11490615 TI - Nitrogen trafficking and recycling through the human bowel. PMID- 11490616 TI - Role of the systemic inflammatory response in the development of protein-energy malnutrition in inflammatory bowel disease. PMID- 11490617 TI - Diagnostic criteria for chronic inflammatory bowel disease in childhood. PMID- 11490618 TI - Nutrition and gene expression. PMID- 11490619 TI - Nutrition versus drug therapy. PMID- 11490620 TI - Use of macro- and micronutrients for nutrition support in inflammatory bowel disease. PMID- 11490621 TI - Enteral nutrition in children. PMID- 11490622 TI - Remission induced by a new specific oral polymeric diet in children with Crohn's disease. AB - We have been able to show that CT3211 is an effective oral treatment in children with active Crohn's disease. It was well tolerated, and there were minimal side effects. At the mucosal site of disease there was macroscopic and histological improvement, together with evidence of downregulation of the pro-inflammatory cytokines IL-1 beta, IL-8, and IFN-gamma. PMID- 11490623 TI - Lipid treatment in inflammatory bowel disease. PMID- 11490624 TI - Potential role of glutamine administration in inflammatory bowel disease. PMID- 11490625 TI - Genetic aspects of inflammatory bowel disease. PMID- 11490626 TI - Monoclonal antibody therapy in inflammatory bowel disease. PMID- 11490627 TI - Immunopathogenesis of inflammatory bowel disease: role of cytokines and immune cell-enterocyte interactions. PMID- 11490628 TI - Nutritional influences on inflammation. PMID- 11490629 TI - Epidemiology of inflammatory bowel disease. PMID- 11490630 TI - Clinical and pathological aspects of inflammatory bowel disease. PMID- 11490631 TI - Diagnostic criteria for inflammatory bowel disease in adults. PMID- 11490633 TI - Periodontal diseases in children and adolescents: I. Aetiology and diagnosis. AB - A number of periodontal diseases, some forms of which can be aggressive, can affect children and adolescents. Plaque is the key aetiological agent but local and systemic factors that can modify the response of the periodontal tissues to plaque may be identified from the history and during examination of the child or adolescent. Periodontal screening should be an integral part of the dental examination of younger individuals. It can be used in conjunction with the history and examination to reach a diagnosis, which then forms the basis for future management. PMID- 11490632 TI - A dental dream. PMID- 11490634 TI - Implant complications and failures: the complete overdenture. AB - The implant-retained overdenture has been advocated as an effective method of restoring the edentulous patient with an appliance that offers significant benefits over conventional prosthetics. However, the success and survival of such appliances have been shown to vary considerably, depending on which jaw is treated: implants supporting overdentures in the maxilla have a significantly higher failure rate. The restoration also requires considerable maintenance, which is both time consuming and costly. The purpose of this paper is to look at common restorative complications and maintenance problems following the placement of dental implants to support an overdenture. A number of implant systems have been reviewed and the results of the authors' clinical experiences with referred and their own cases are reported. PMID- 11490635 TI - Treatment planning for the problem patient: restorative, ethical, legal and psychological perspectives. Case I: Emma. AB - 'Emma' is the first of a series of four articles that aims to look at some of the problems patients may present with when they visit their dentist. Each article starts with a brief scenario about the hypothetical patient and his/her clinical problems. As well as the clinical problem, each case also raises ethical, legal or psychological problems. All of these issues are discussed and possible management strategies and treatment options described. PMID- 11490636 TI - The assessment of the endodontically treated tooth. AB - This paper reviews the assessment of success and failure following endodontic treatment and highlights the fact that many referrals for specialist management of endodontic failure do not take into consideration established guidelines in this important area. PMID- 11490637 TI - Snoring, sleep apnoea and the role of dental appliances. AB - This article describes the problems of snoring and obstructive sleep apnoea, together with an outline of treatment options. The Glasgow approach, whereby patients are investigated at a sleep clinic and a custom-made mandibular advancement device is made in semi-soft material, is also described. We have demonstrated the acceptability and effectiveness of a simple appliance in patients with varying dental states, some with simple snoring and some with mild to moderate sleep apnoea. Our experience relates to around 260 patients, extending over a period of 4 years with good success. The simple intraoral device is recommended as a first line of approach for patients with problem snoring. PMID- 11490638 TI - Multiple myeloma presenting as a painful mandibular swelling: a case report. AB - Multiple myeloma is a disease characterized by monoclonal proliferation of plasma cells, the most differentiated stage of B-cells. Primary manifestation of multiple myeloma in the jawbones is rare. In the case reported here, a 29-year old woman who presented with a right mandibular swelling after extraction of a mobile painful tooth turned out to have multiple myeloma. Current diagnostic criteria and management strategies of the disease are discussed. PMID- 11490639 TI - Does orthodontics damage faces? PMID- 11490640 TI - Lateral thinking required! PMID- 11490641 TI - Lanarkshire children's tooth decay halved. PMID- 11490642 TI - Rural dentistry is what you make of it. PMID- 11490643 TI - The journey back. Part Four: The central regional boards. PMID- 11490644 TI - The effect of extensive consumption of soda pop on the permanent dentition: a case report. PMID- 11490645 TI - What's new at the dental school? PMID- 11490646 TI - Dental management of HIV/AIDS patients. PMID- 11490647 TI - Sleep apnea: what's a dentist to do? PMID- 11490648 TI - Articaine: the new, old anesthetic. PMID- 11490649 TI - The state of the state: dental practice in 2000. PMID- 11490650 TI - A crisis in the community. PMID- 11490651 TI - Joint venture versus outreach: a financial analysis of case studies. AB - Medical centers across the country are facing cost challenges, and national commercial laboratories are experiencing financial declines that necessitate their capturing market share in any way possible. Many laboratories are turning to joint ventures or partnerships for financial relief. However, it often is in the best interest of the patient and the medical center to integrate laboratory services across the continuum of care. This article analyzes two hypothetical joint ventures involving a laboratory management agreement and full laboratory outsourcing. PMID- 11490652 TI - Physicians and compliance: developing a system that works. AB - This article will describe the development of a method to educate physicians and other health-care providers about the components of the Model Compliance Plan for Clinical Laboratories. It incorporates adult learning principles through the use of academic detailing and other educational tools. The improvement process and indicator data used by our health-care facility is outlined. Since the release of the Model Compliance Plan for Clinical Laboratories in 1997, it has been an ongoing challenge to meet its guidelines and develop processes to ensure compliance of medical necessity documentation and submission of accurate claims for Medicare reimbursement. Most laboratories have the appropriate policies and procedures in place to achieve effective compliance programs. However, the existence of a policy does not ensure compliance. In our organization, Mercy Medical Center-North Iowa, Mason City, Iowa, baseline data demonstrated the need for process improvement in the area of physician office compliance. Often, advance beneficiary notices or notices of non-coverage were nonexistent or incomplete; medical necessity documentation for the requested tests was inconsistent; and patient demographic information was incomplete. As a result, Medicare could not be billed and laboratory write-offs were at an unacceptable level. We will outline some of the compliance issues currently facing laboratories and describe the systems approach used to improve physician compliance in our system. PMID- 11490653 TI - EXCEL gradebooks: an EXCELlent strategy for saving teacher time. AB - Time ultimately is the most precious commodity across professions. In the teaching profession, educators put in pre-class development time, class contact time, and post-class grading and grade calculation time (as well as other administrative time). End-of-term grade calculation can be a massive, time wasting headache. If teachers develop electronic spreadsheets that track attendance and use formulas to do the final calculations automatically as data is entered into the spreadsheets, then teachers can nearly eliminate end-of-term number crunching and use that time for other endeavors. PMID- 11490654 TI - Successful community-based laboratory services program for long-term care facilities, Part 2. AB - Hagerstown Medical Laboratory, Inc., located in Hagerstown, Maryland, has an outreach program that currently provides laboratory services to 52 long-term care facilities. Part 1 of this series, published in the May/June 2001 issue of Clinical Leadership & Management Review, discussed general organization, staffing, and safety issues for their Nursing Home Program. Part 2 relates their experience with contracts, fees, and reimbursement. PMID- 11490655 TI - Key elements of government contracting: an untapped resource for hospital outreach revenues. AB - Historically, hospitals hesitate to compete against commercial laboratories for public sector and managed care market opportunities that would increase a hospital's revenue base. Hospital laboratories perceive that they cannot compete in the scope and complexity of testing, the service requirements, or commodity pricing strategies of commercial laboratories. Since 1997, the Department of Pathology Laboratories at Virginia Commonwealth University Health System's Medical College of Virginia Hospitals and Physicians has been awarded major contracts with government agencies and managed care companies to provide laboratory services for local and regional networks. This article reviews the key elements and ongoing price and service strategies in a sequential, easily followed format that hospital outreach programs can use as a resource when expanding their sales effort into the public sector market. Readers instantly should realize the immediate financial rewards from public sector contracts that offset the industry's declining reimbursements from third party payers. PMID- 11490656 TI - Understanding and using organizational politics. PMID- 11490657 TI - Genetic test results and information systems. PMID- 11490658 TI - Common cent$ 1: One-armed economists and the invisible hand. AB - This article is the first in a series called Common Cent$. There is a need for leaders and managers to have a basic understanding--Common Cent$--of elementary economics. The limited, retrospective view of the accountant must be supplemented by the broader, prospective view of the economist. The limits and scope of economics are defined. The First and Second Fundamental Theorems of Welfare Economics are introduced. The mythology behind the mechanism of action of Adam Smith's Invisible Hand is dissected, and the mechanism of the free market is explained in terms of the effect of marginal cost on net market efficiency. The apparently simple case of the effect of legislating a minimum wage on a free market is discussed. This provides an example of the real-world complexity of economies and of applying economic concepts to the business world. PMID- 11490659 TI - The same side of the fence: Part 1. Managers, educators, and health-care administrators all depend on each other. PMID- 11490660 TI - Meeting the challenges of customer retention and staff retention. PMID- 11490661 TI - Establishing and maintaining efficient courier services. AB - Your couriers are your link to your outreach customers. They represent your facility to and carry information (and, of course, specimens) from the communities you service. When establishing your outreach program, who is in the driver's seat and how specimens will be transported are primary concerns. Cost conscious laboratorians rely on a variety of courier options, from transporting specimens on foot using refrigerated Styrofoam nylon packs to calling taxis to contracting with drivers to having a dedicated arrangement with in-house employees. Often, the expense and bother of maintaining a fleet of cars and drivers, probably most desirable in terms of control and standardizing processes, forces laboratory managers to seek alternatives that may not sound practical or reliable, but that actually do work quite well. Whatever courier route you choose, you should emphasize to your carriers the respect you have for the precious cargo they carry. Of course, there must be no doubt about specimen integrity, carefully and conscientiously maintained from its source to you. But the integrity of your organization is also in the hands of those who accept the responsibility of bringing you all of those potential questions to be answered. Your livelihood depends on it. Just be as certain as possible that your customers want to celebrate--not shoot--your messenger. PMID- 11490662 TI - Patient confidentiality: privacy versus clinician access. PMID- 11490663 TI - Encouraging professional service as a staff development tool. PMID- 11490664 TI - [Medical manpower in Switzerland: past and future trends]. AB - In 1966, an official report predicted a 3-decades period of shortage of physicians in private practice in Switzerland. However a demographic analysis of the trends of medical students provided a totally different picture, as early as 1971. As a matter of fact, the ratio of doctors in private practice per 100,000 population rose to 195 in 2000 from 88 three decades earlier. In 2020, Switzerland will have a ratio of 240-270, whatever the decisions made today. Will such a tremendous increase introduce a damaging surplus of private practitioners? Medical progress, population aging, increase in the per capita demand for health care and other factors as well give raise to an other conclusion, much more optimistic. PMID- 11490665 TI - [Factors affecting regional distribution of French physicians]. AB - Regional maldistribution of French physicians--as measured by the physicians/population ratio--has been reduced during the period 1985-2000. The numerus clausus policy (the medical student intake is determined every year by the government) which favors to some extent the underserved regions has contributed to the process of equalization. However this is applicable only to GPs whose the training at post graduate level is carried out in their own region. The students who are admitted to specialty training have a greater regional mobility and have not strongly contributed to the regional equalization process. After their training, medical doctors may migrate from a region to another. The migratory flows have increased the regional maldistribution. On the other side, the retirement flows, which have not had the same importance in all regions, have strengthened the equalization process. PMID- 11490666 TI - [Regional disparities in the distribution of French physicians in private practice (1998-2008)]. AB - Forecasts are showing that the gap among French regions as concerns supply of doctors in private practice will narrow during the decade 1998-2008. However the trend will be more pronounced for GPs. Unequal supply of specialists at regional level will remain. For GPs and specialists as well, unequal supply is more important within a region that between various regions. The feature will remain during the coming decade. PMID- 11490667 TI - [Regulating the distribution of health professionals in Germany, Belgium, the United Kingdom, Quebec and the United states: monograph study]. AB - The Province of Quebec and the United Kingdom have both a strong system of medical manpower regulation. Growth of the number of doctors--and consequently of health expenditures--is kept under control. Shortage of supply of health services increases the queuing process. In the two countries, the main problem is not how to curb health costs but to cope with the unmet demand. In Germany, the control of medical school intake is very weak. However, agreement of young doctors by the Sickness Fund is strictly controlled, the Fund having the right to close the list of "agreed physicians" in areas or specialties where doctor surplus is observed. In Belgium, a numerus clausus system is established very recently for medical school intake. In the USA, the health care system is traditionally regulated by the market forces. The strong increase of health expenditures during the recent decades has shifted the power from the medical profession to the paying bodies (HMO, MCO...) who are now supporting the emergence of new health personnels, less costly. PMID- 11490668 TI - [Emigration of Quebec physicians: motivation for departure and return]. AB - Since the 80's, outmigration of physicians from Quebec is steadily increasing. About 46 percent of outmigrating doctors explain their move by factors related to their occupational life (higher income, greater opportunity in the academic career, larger amount of resources devoted to the health care system). Nearly 40 percent relate their decision to personal factors (greater job opportunity for their wife/husband, quality of family life...). The factors linked to the context of the receiving place (political climate, linguistic regulations, income tax level...) play a minor role on the migration decision. As concerns the returning physicians, 80 percent explain their decision by personal factors. The factors linked to the occupational life have a lower role. It appears therefore that doctor outmigration from Quebec is not directly determined by manpower policies adopted by the Province during the last two decades, except the policies directly linked to the income level of professionals. PMID- 11490670 TI - The psychotherapy of the schizophrenic patient: an interview with Dr. Silvano Arieti. Interview by Dr. Milton M. Berger. PMID- 11490669 TI - [Beyond the statistics: why are Quebec physicians emigrating?]. AB - During the period 1986-1999, about 16% of the annual average number of active physicians in Quebec have left for the US or an other Canadian province. The absolute number was 2367. During the same period, 661 would have returned to Quebec. The majority of the outmigrating doctors were graduated from the anglophone university of the Province (although 44 percent are francophone) whereas the majority of the returning doctors were graduated from the 3 francophone universities of the Province. As a matter of fact, 78 percent of the returning doctors are francophone. The migration decision is equally linked to post-graduate training and occupational opportunities and job satisfaction. This feature should be related to an other: those who have returned to Quebec have had a geographic and occupational mobility level far lower than those who have not returned. Finally, it is noteworthy that there are significant differences between the doctors emigrating to the US and their colleagues going to an other Canadian province. PMID- 11490671 TI - Franz Alexander: a unique outstanding pioneer. PMID- 11490672 TI - Androgyny in Indian art and culture: psychoanalytic implications. PMID- 11490673 TI - Idealization and reverence. PMID- 11490674 TI - Mourning and adaptation following the death of a parent in childhood. PMID- 11490675 TI - Transference and countertransference contributions toward understanding the phenomenon of institutionalization of schizophrenic patients. PMID- 11490676 TI - Wounded healer: the impact of a therapist's illness on the therapeutic situation. PMID- 11490677 TI - Facing mortality while treating patients: a plea for a measure of authenticity. PMID- 11490678 TI - Psychoanalytic perspectives on theories regarding the development of antisocial behavior. PMID- 11490679 TI - Countertransference and ethnicity: the analyst's psychic changes. PMID- 11490680 TI - Adolescent trauma in Japanese schools: two case studies of Ijime (bullying) and school refusal. PMID- 11490681 TI - Psychiatric Darwinism = survival of the fittest + extinction of the unfit. AB - This article is a critical analysis of the American Health Security Act of 1993. Although AHSA was soundly defeated when first proposed, parts of it have been enacted into law in 1996, with the prospect of further piece-meal enactments in the future. It includes matters of fundamental importance to American mental health practitioners, to vulnerable citizens with psychiatric disorders, to their families, and to their few champions in medicine and law. Utilitarianism is the unstated philosophical substructure of AHSA and its legislative progeny, i.e., whatever cuts medical costs and saves money is good. The author delineates AHSA's mental health entitlements and limitations of in-patient, out-patient, and other patient care. She enumerates a dozen major imperfections and dangers of this mental health law, especially its medical utilitarianism emphasizing outcomes and quality of life. Dr. Cosman argues that medical cost, outcome, quality of life, and managed competition threaten the essential liberties and the lives of older persons, persons who are chronically ill, fatally ill, and most particularly those who are mentally impaired. She concludes that if limited money, medicine and time are invested only in inevitable medical success, then America's medicine by its medical law will be Medical Darwinism encouraging survival of the fittest by requiring extinction of the unfit. PMID- 11490682 TI - An outsider's view of Dutch euthanasia policy and practice. AB - This article provides a critical analysis of Dutch euthanasia policy and practice. The research benefited from twenty-eight interviews conducted in the Netherlands during the summer of 1999 with some of the leading figures who dictate the decision-making process and take an active part in the debates. The discussion begins with a review of the two major Dutch reports on euthanasia and the conflicting views and interpretations offered by the literature. Next, I provide some data about the interviews, and then analysis indicating that the Dutch Guidelines on the policy and practice of euthanasia do not provide ample mechanisms against abuse. I argue that the Dutch Guidelines are insufficient, do not provide adequate control over the practice of euthanasia, and that the entire policy should be revised and made more coherent and more comprehensive. PMID- 11490683 TI - Robert William Latimer v. Her Majesty The Queen: in the Supreme Court of Canada. PMID- 11490684 TI - A perennial controversy. PMID- 11490685 TI - Proverbs shine light on ergonomics. PMID- 11490686 TI - Second-hand smoke may cause caries in children. PMID- 11490687 TI - Tea fights cavities, reduces plaque. PMID- 11490688 TI - Oral, tongue cancer rates rise among young Americans. PMID- 11490689 TI - How and why politics affect dentistry. AB - Dental practices are under scrutiny every day. The dynamics of the public, the media, the lawmakers, the regulators, and other special-interest groups create endless possibilities for influence over a practice and continue to challenge a dentist's ability to provide quality dental care to patients. This article describes examples of laws and regulations affecting dentistry and the impetus for them, whether real or perceived. PMID- 11490690 TI - Proposition 65 in the dental office. AB - In the mid-1990s, dentistry became included in enforcement activity for Proposition 65, the Safe Drinking Water and Toxic Enforcement Act of 1986. This year, 80 dental offices were served with 60-day notices for failing to post Prop. 65 warnings. The California Dental Association has been inundated with inquiries on what dental offices should be doing with regard to Prop. 65 requirements. This article provides a brief history of Prop. 65 and answers the questions most frequently asked of CDA. PMID- 11490691 TI - Dentistry, amalgam, and pollution prevention. AB - California has issued fish consumption advisories because of mercury in lakes, reservoirs, creeks, rivers, and bays. Mercury in these waterways leads to the formation of methylmercury, which is toxic and bioaccumulative. Dental practices and other health care settings contribute a portion of this mercury. Government agencies are implementing programs to reduce mercury pollution. Dentists can reduce their contributions by implementing best management practices. They may also consider using pretreatment technologies as more information becomes available about their use and effectiveness. PMID- 11490692 TI - Monitoring external forces that affect the practice of dentistry. AB - Dealing with the varied and often changing requirements and regulations that affect every dental practice can be a daunting task for the dentist and dental staff. Knowing how and where to access accurate and timely information provides relief for the busy dental professional. This article explores the impact of regulatory mandates and government agency standards on dentistry and provides suggestions for simplifying the search for reliable information. PMID- 11490693 TI - The ethical responsibility of professional autonomy. AB - Dentistry has historically been practiced autonomously, and many dentists place a high value on professional independence. This article outlines the conceptual basis for professional autonomy and asserts that dentists can retain independence only by aligning values with patients and remaining trustworthy in the eyes of the public. PMID- 11490694 TI - A comparison of the cost effectiveness of pressure-indicating materials and their ability to detect pressure areas in complete dentures. AB - Pressure areas on the fitting surfaces of dentures can cause great discomfort and pain to denture wearers if not detected and removed. Pressure-indicating materials are commonly used to detect these areas, and several commercial varieties are available, but these tend to be expensive. The cost effectiveness of these materials has not been investigated, nor has this been linked to their efficacy and ease of use. The aim of this study therefore, was to compare the different pressure-indicating materials available commercially with that of a home-made paste. An assumption of efficacy was made by the number and size of pressure areas revealed, by taking standardised photographs and analysing the images. User friendliness was determined by the time taken to mix, apply and remove the material. Cost was determined by a cost per unit calculation based on the average or minimum quantity required for each material. The results indicated that a home-made paste made of equal quantities of hand lanolin (BP) and zinc oxide powder was not only the most effective, but was also the cheapest, being only 3% of the cost of the most expensive of the materials. PMID- 11490695 TI - Wastage of a silicone impression material in a general practice setting: a comparison between hand and automixing methods. AB - A practice-based, randomised, controlled trial has been undertaken to investigate the wastage of impression material when recording full arch one-stage, polyvinyl siloxane impressions using four techniques: putty and automix light viscosity material, putty and automix regular viscosity material, putty and tubed light viscosity material, and putty and tubed regular viscosity material. A total of 100 maxillary impressions, comprising 25 recorded using each of the four techniques, were obtained according to a predetermined scheme for randomisation. All material wasted in dispensing, mixing and applying each impression was determined by repeated weighing. Statistical comparisons of the mean weights of material wasted for each type of impression revealed that significantly (P < 0.01) less wastage occurred with the automix techniques. It is concluded that automix impression techniques may, in more extensive studies, be found to be more cost effective than tubed impression material techniques when recording impressions of the type investigated. PMID- 11490696 TI - A subjective assessment of pain and swelling following the surgical removal of impacted third molar teeth using different surgical techniques. AB - Pain and swelling are two of the most common problems experienced by patients who have undergone surgical removal of impacted third molars. The purpose of this research project was to ascertain which of two surgical techniques was judged by patients to cause the least pain and swelling. The survey comprised 20 patients with bilaterally symmetrical impacted third molar teeth. In each patient, the third molars on one side were removed using a standard mucoperiosteal flap, while on the opposite side, a smaller access incision was used. The results of this survey show conclusively that when a small incision was used, with minimal reflection of the mucoperiosteum, the subjective evaluation of patients is that there is significantly less postoperative pain and swelling than when the larger standard incision is used. PMID- 11490697 TI - Gardner's syndrome--the importance of early diagnosis: a case report and a review. AB - Gardner's syndrome (familial polyposis coli) is a genetic condition characterised by colonic polyps that carry a 100% risk of malignancy if untreated. Early diagnosis, in which an astute and knowledgeable dentist can play an essential role, is therefore of paramount importance. The presence of multiple unerupted teeth provides the dentist with a major pointer to the possible presence of this disease, which may be provisionally diagnosed on the detection of two other obvious features, namely osteomas and cutaneous lesions. This fact is clearly illustrated by the accompanying case report. Extracolonic manifestations increase the morbidity and make treatment more difficult. The presence of large desmoid tumours may be a serious complicating factor. Gardner's syndrome may be accompanied by adenomas of the stomach and duodenum and very rarely by malignant tumours of the central nervous system. Regular surveillance by means of colonoscopies must be carried out on all individuals suspected of having the disease in order to implement timeous life-saving prophylaxis. PMID- 11490698 TI - Risks and values in treatment decision making. AB - The 1984 Oral Health Strategy defined oral health as a subjective state rather than an objective one. It also made clear that oral health was far more than the mere absence of oral disease. Healthcare outcomes can therefore only be evaluated by the individual receiving healthcare rather than by those who provide it. The task of the healthcare provider is to determine the probability of achieving particular health outcomes. Weighting the value of an outcome by the probability with which it will occur can inform decision-making. The decisions which result from using such a system are sometimes counter-intuitive but ensure that the patient's values are fully incorporated into the decision-making process. This is particularly important in cases where the problem is aesthetic rather than pathological. Patients' evaluations of health outcomes are also known as health state utilities. Even if only considered implicitly, use of utility weighting ensures that patient consent is informed consent and encourages communication and understanding between patient and practitioners. PMID- 11490699 TI - Oral signs and symptoms as predictors of salivary gland hypofunction in general dental practice. AB - OBJECTIVE: To evaluate the signs and symptoms of oral dryness as predictors of salivary gland hypofunction (SGH) in general dental practice. DESIGN AND SETTING: Prospective study recruiting adult patients attending five general dental practices in Merseyside in 1999. MATERIALS AND METHOD: Patients were screened for subjective symptoms of oral dysfunction and clinical signs of oral dryness. Patients with oral symptoms or signs of SGH were invited to undergo sialometry. Results were analysed using multiple logistic regression. RESULTS: 1103 patients were screened for signs and symptoms of oral dryness, 115 reported continuous xerostomia, of these 65 were also clinically (subjectively) assessed as having a dry oral mucosa. One hundred and one patients attended for sialometry and 73% of these had objective evidence of SGH. Neither the patients' complaints of oral dryness or the assessment of dryness of the oral mucosa were significant predictors of SGH. CONCLUSIONS: Symptoms of oral dysfunction and clinical signs of oral dryness were not significant predictors of SGH in dental practice. PMID- 11490700 TI - To prescribe or not to prescribe. PMID- 11490701 TI - Accessing the literature: using bibliographic databases to find journal articles. Part 1. AB - Research in primary dental care, recertification, continuing professional development, lifelong learning, peer review and quality healthcare are all informed by the published literature. Dental practitioners can find out about reliable and up-to-date information available in the published literature by searching bibliographic databases. Published in two parts, this article describes the databases relevant to clinical dental practice and explains the generic skills required to search them effectively, focusing on MEDLINE, the database most relevant for the majority of dental practitioners, which is freely available via the World Wide Web (WWW). The article differentiates between sensitivity (maximum recall) and specificity (relevance of recall), and suggests how to identify a manageable number of relevant citations, how to save the citations, and how to obtain the full text. In part 2, the article concludes by alerting readers to some of the limitations and pitfalls of database-searching. PMID- 11490702 TI - An evaluation of attitudes of Midlands female dentists to general dental practice: will it impact upon the future delivery of primary dental care? PMID- 11490703 TI - Visual standards in a sample of dentists working within Scotland. AB - INTRODUCTION: This study aims to measure the visual acuity of practising dentists and to establish the average time elapsed since their last routine eye examination. MATERIALS AND METHODS: Distance visual acuity, near visual acuity, convergence, accommodation, contrast sensitivity, heterophorias and stereoscopic vision were measured in a diverse group of 46 dentists practising in Scotland. RESULTS: The results indicated that distance visual acuity was at least 6/6 for all but one of the subjects. All had acceptable near visual acuity. Accommodation followed the expected age-related trend of decreasing with age. No subjects had a definitely abnormal contrast sensitivity score although four were in the borderline zone. No subjects had a horizontal squint but two had a significant vertical phoria. Three subjects had no stereoscopic vision and one subject had a very high threshold level. All other subjects were able to see stereoscopically. Of the subjects 57% had been for an eye examination within the previous two years, with the range being from less than one month to 17 years. CONCLUSION: The eyesight of this group of clinicians exceeds generally accepted normal population values. Although the majority of the clinicians had attended for an eye examination within two years, the large range of time since last attendance is of concern and regular eye examinations are to be encouraged. PMID- 11490704 TI - Looking back--Dean's address. AB - General dental practitioners have been part of The Royal College of Surgeons of England since the LDS RCS examinations in 1860. For 87 years, it was the general dentists who were part of the College. Later, the political necessity of introducing hospital dentists to the National Health Service changed the ethos for dentists with the formation of the Faculty of Dental Surgery in 1947. Secondary care dentists became entrenched within The College at the expense of the majority of those who practiced dentistry to the same level as secondary care dentists. It took a further 45 years before general practitioners could return to their home. The Faculty of General Dental Practitioners (UK), within a short space of time, has become the largest Faculty for dentists within any of the Royal Colleges. This article includes excerpts from the Dean's address at the Ceremony of Presentation of Diplomats held on 3rd March 2001 (see page 75). It unveils the accidents of history which formulated a Royal College, the perseverance of committed dentists which took them into the fold of The College. It describes the beginning of the Faculty of Dental Surgery and unravels the story of the Advisory Board in General Dental Practice which later became the Faculty of General Dental Practitioners (UK). The next article 'Looking Forwards' will describe a way forward for this Faculty, which has grown up all too quickly in a rapidly changing world. PMID- 11490705 TI - The development and use of a triage protocol for patients with dental problems contacting an out-of-hours general medical practitioner cooperative. AB - PURPOSE OF THE STUDY: The number of patients contacting general medical practitioner (GMP) out-of-hours services with dental problems is perceived to be a significant problem by the medical profession. This study was undertaken to design and pilot a triage protocol that could be used by non-dental staff to refer callers with dental complaints for appropriate treatment. BASIC PROCEDURES: A triage protocol was designed to address the patient conditions considered to be relevant to emergency dental care. The triage protocol was piloted for three months at a GMP cooperative in North Wales, which provided an out-of-hours service for 61 GMPs. Baseline data were collected for three months prior to the introduction of the triage protocol. MAIN FINDINGS: The study showed that the number of dental callers contacting the out-of-hours service was not as great as GMPs imagined, and was similar to other studies. The introduction of the triage protocol resulted in a rise in the number of callers asked to call the general dental practitioner (GDP) support line. The number of unregistered patients given the name of a dentist also increased. The need to make a second telephone call seemed not to affect patient satisfaction since when patients were later questioned on their experience, those who had used the service since the introduction of the triage protocol were more likely to be satisfied with the service. PRINCIPAL CONCLUSIONS: This study provides some evidence that a triage protocol can be used successfully by non-dental staff to sift requests for out-of hours emergency dental care. PMID- 11490706 TI - Advances in dental public health. AB - Dental public health has been defined as 'the science and art of preventing oral diseases, promoting oral health and improving the quality of life through the organised efforts of society'. Dental practitioners most often have the oral health of individual patients as their primary focus but the aim of public health is to benefit populations. Early developments in dental public health were concerned largely with demonstrating levels of disease and with treatment services. With greater appreciation of the nature of oral health and disease, and of their determinants has come recognition of the need for wider public health action if the effects of prevention and oral health promotion are to be maximized. PMID- 11490707 TI - Toward science-based practice: a charge from the US Surgeon General. PMID- 11490708 TI - [Surgical tactics in complicated forms of acute purulent pyelonephritis in pregnancy]. AB - Out of 3000 pregnant women observed in 1986-1999, pyodestructive forms of pyelonephritis were diagnosed in 120 women. They were treated surgically. The policy of the treatment depended on the degree of pyoseptic intoxication, severity of toxic affection of the parenchymatous organs, severity of pyodestructive changes in the kidney. Plasmapheresis expanded indications to organ-saving operations, effectively and quickly arrested purulent intoxication. Various organ-saving operations have been successfully performed in 77(64.1%) pregnant women. In 12 patients nephrostomy was uneffective. In these patients a second nephrectomy was made. Primary nephrectomy was made in 37 cases of life threatening pyoseptic intoxication. Hard to treat bilateral pyodestructive forms of pyelonephritis were detected in 15(12.5%) women. Bilateral nephrostomy with cleaning of purulent infection foci was effective in 9 patients. It is shown that early surgical detoxication of the most affected kidney prevents development of extended pyodestructive alterations in the contralateral kidney and avoid surgical intervention. PMID- 11490709 TI - [Pharmacological and cost effectiveness bases of the use of categel and categel S [correction of F] in urological practice]. AB - Preparations catedgel and catedgel S made in Austria (Montavit) was tried in Moscow hospital N 50. Categel is a sterile gel of methylcellulose with 2% lidocain and 0.05% chlorhexidine, catedgel S contains the same components but lidocain. Categel significantly reduces the risk of infectious-inflammatory complications after endourological manipulations, improves endoscopic diagnosis and makes some manipulations less painful. Comparative pharmacological cost effect assessment of categel S and glycerine effects in prostatic transurethral resection. Categel was found 2.11 times more effective. It also improves quality of life of the patients. Categel can be recommended for wide use in urology. PMID- 11490710 TI - [Relationship between biological variants of U. urealyticum and clinico morphological manifestations of urogenital inflammation in experimental animals]. AB - Dependence of clinicomorphological changes in urinary and sexual organs of rats on biological variants of U.urealyticum (biovar Parvo or T960) used in experimental modelling of ureaplasmic infection by two variants: a) introduction in the sterile conditions of culture U.urealyticum (10(4)-10(6) U/ml) directly into the bladder by catheterisation b) infection of female anogenital region with culture U.urealyticum (10(6) U/ml) for 5 days and keeping healthy males and females together. Clinicomicrobiological and morphological evaluation of the urinary and sexual organs of the rats was made 3, 7, 14 and 30 days after the infection. Features of the inflammation were specified with reference to a certain biovar of U.urealyticum. Identity of U.urealyticun biovars used for infection and isolated from the urogenital organs of the test animals 14 and 30 days after the infection was confirmed microbiologically. PMID- 11490711 TI - [Role of thromboxane A2 (B2) in the pathogenesis of stress proteinuria in healthy persons]. AB - Analysis of published data and the author's own findings indicate that processes coursing in the miners of deep coal mines during hard physical work lead to hemodynamic and biochemical shifts, causing decreased production of thromboxane A2 in the kidneys and involvement of the proximal canaliculi. This, in turn, results in increased protein filtration and decrease reabsorption, which eventuates in stress proteinuria. PMID- 11490712 TI - [Connective tissue dysplasia as one of the possible causes of urinary incontinence in women with genital prolapse]. AB - Urinary incontinence (UI) in patients with genital prolapse is not always related to stress. It may result from dyslocation of the vesicourethral segment. While genital prolapse in young patients after a single normal delivery is common manifestation of connective tissue dysplasia (CTD) at the level of the reproductive system, 51.8% cases of UI may be of neurogenic origin and present as dysfunction of the vesicourethral segment and pelvic diaphragm. Unstability of the bladder and urethra in these cases may represent CTD at the level of the pelvic organs. PMID- 11490714 TI - [Comparative analysis of long term results of modified transcutaneous urethro cysto-cervico-pexy with vaginal flap in the treatment of stress urinary incontinence in women]. AB - The authors present a comparative analysis of long-term results of transcutaneous urethrocystocervicopexy (TU) with a vaginal graft according to O. B. Loran-D. Yu. Pushkar and its modification. The former method provided good 2-year results in correction of urinary stress incontinence (USI) in 84.8% of cases, satisfactory and unsatisfactory results were achieved in 6.5 and 8.7%, respectively. In modified TU the results were good, satisfactory and unsatisfactory in 89.7, 6.9 and 3.4%, respectively. This modified method of surgical USI treatment in women allows to create individual degree of tension of pubovaginal ligatures in each female, provides optimal anatomic correlations of the urinary bladder and urethra in TU, prevents obstruction and induced by it disorder in urine evacuation from the bladder in the postoperative period, reduces duration of catheterization of the bladder up to 1-2 days and hospital stay to 4.5 days, results in normal physiological urination. PMID- 11490713 TI - [Intestinal plasty of the ureters in the treatment of acquired obstructive ureterohydronephrosis]. AB - 222 patients with obstructive ureterohydronephrosis aged 14 to 65 years were treated surgically. They have undergone nephroureteroectomy (n = 7), direct ureterocystoneostomy (n = 84). In 134 patients 156 ureters were replaced by an iliac segment (unilaterally in 102 cases, bilaterally in 50 cases, in 4 cases of ureterohydronephrosis of a single kidney). In addition to routine diagnostic methods for ureterohydronephrosis, intraoperative electroureteromyography was used. By intensity of ureteral biopotentials, the borders of nonfunctioning zone of the ureter were estimated. Normal bioelectricity was found 40 to 60 mm down and up from the pathological focus, respectively. The presence of structural changes in the ureters' wall within this zone was proved by morphological investigations. Indications for intestinal plastic surgery of the ureter were the following: recurrent ureteral strictures, ureteral fistula, long strictures of the ureter, prepelvic strictures of the ureter, multiple strictures (36, 21, 69, 16 and 7 cases, respectively). All the operations were performed in one stage through the abdominal cavity with a single intestinal transplant. The above plastic surgery restores both patency of the urinary tracts and transportation of uric bolus to the bladder. Postoperative course depends mostly on the initial condition of the patient and function of the kidneys. Postoperative lethality was 0.45%. Intestinal plastic surgery of the ureter is a valuable alternative to nephroureteroectomy in bilateral pathology of the kidney and in ureterohydronephrosis of a single kidney. PMID- 11490715 TI - [Prostamol-uno in the treatment of benign prostatic hyperplasia]. PMID- 11490716 TI - [Differential diagnostic value of prostate-specific antigen in prostatic hyperplasia]. AB - Diagnostic value of prostate specific antigen (PSA) was studied in 163 patients with prostatic hyperplasia. As PSA is not a cancer-specific test, it is recommended to obtain biopsy in patients with prostatic hyperplasia and PSA > 4 ng/ml to diagnose prostatic cancer early. Methods raising PSA specificity for differential diagnosis of prostatic hyperplasia are not absolutely reliable. PMID- 11490717 TI - [Clinical significance of the determination of R-proteins in the ejaculate of patients with chronic prostatitis]. AB - Products of cell receptor (R-proteins) catabolic lysis were measured in ejaculate of 121 patients with chronic prostatitis and 15 healthy males. R-protein levels were higher in the patients (U = 172.5; p = 0.047). When immunological indices of the ejaculate (IgA, sIgA, IgG, IgM, interleukin-8) were measured, it was found that in patients with chronic prostatitis there is a positive correlation between concentration of interleukin-8 and content of R-proteins (r = 0.43; p = 0.028). Positive correlation also exists between R-protein content and prostate volume (r = 0.24; p = 0.034) while negative one--between R-proteins concentration and size of the hypoechogenic sites in the prostate (r = -0.25; p = 0.039), between R proteins in the ejaculate and serum level of fibrinogen B (r = -0.25; p = 0.021). It is inferred that elevation of ejaculate R-protein level occurs in alteration of the prostate. The test for R-proteins in ejaculate can be helpful in detection of early stages of chronic prostatitis. PMID- 11490718 TI - [Immune disorders in congenital hydronephrosis complicated by obstructive pyelonephritis]. AB - Congenital hydronephrotic transformation complicated by obstructive pyelonephritis with intact renal function in children exhibited association with some antigens of the major histocompatibility system and their combinations, changes in immunity and nonspecific resistance. Marked and stable shifts in immune reactivity give grounds for inclusion of immunomodulating drugs in combined treatment of the above patients. PMID- 11490720 TI - [Terminal renal failure in children]. PMID- 11490719 TI - [Acute urinary retention caused by urethral polyp]. AB - A polyp of the prostatic part of the urethra is a rare pathological finding. A 20 year-old male visited our clinic with acute urinary retention. Ultrasound investigation, x-ray and endoscopic examinations revealed a polyp of the prostatic urethra. The polyp was resected by Ho-YAG laser. The resection resolved the urinary retention. The histopathological diagnosis was fibroepithelial polyp. PMID- 11490721 TI - [Methods of diagnosis and treatment of acute diseases of organs of the scrotum]. PMID- 11490722 TI - [Effects of impaired urodynamics and contractile function of the upper urinary tract on elimination of stone fragments after extracorporeal lithotripsy]. AB - The authors analyze effects of amplitude, tone and frequency of ureteral contractions on elimination of stones fragmented after extracorporeal shock-wave lithotripsy (ECSWL) in 30 patients with concrements in the kidneys and ureters. Before ECSWL the patients had indications for renal pelvis cathetarization. The measurements were made with multichannel impedance ureterography. According to initial dilatation of the upper urinary tracts (UUT), the patients were divided into 3 groups. It is shown that urodynamic UUT disorders determine effectiveness of ECSWL in urolithiasis patients: the greater is the dilatation the more probable are unsatisfactory results of ECSWL. Ureteral contractile activity is essential for rehabilitation in early postoperative period after ECSWL in patients with unrevealed UUT dilation: high amplitude of ureteral contractions in low tone is good for the fragments elimination, while high tone may result to the fragments ejection into the kidney. Low contractility may contribute to development of stone path. In patients with pronounced UUT dilation ureteral contractility is less important for ECSWL effectiveness than the wall tone and retrograde ureteral peristaltic movements. ECSWL results are better in patients with low tone of UUT wall. High UUT tone, especially in combination with retrograde peristaltic ureteral contractions registered prior to ECSWL indicates high probability of residual stones and additional fragmentation procedures irrespective of initial UUT dilation. PMID- 11490723 TI - Pennsylvania dental workforce. Statistics and trends. PMID- 11490724 TI - Mobile dentistry in the nursing home. PMID- 11490725 TI - Preparing for the future: priorities in dental education at Penn. PMID- 11490726 TI - Refocusing its vision: the University of Pittsburgh School of Dental Medicine. PMID- 11490727 TI - Dentistry at Temple University--the tradition continues. PMID- 11490728 TI - Dudley and his friends "Sport a Winning Smile" during National Children's Dental Health Month! PMID- 11490729 TI - Survey reveals practices, attitudes and preferences of major Pennsylvania employers about dental benefits. PMID- 11490730 TI - Legislated to death. PMID- 11490731 TI - First half of 2001: "Action Packed" legislative session. PMID- 11490733 TI - We answer your questions about Smilemichigan.com. PMID- 11490732 TI - Release of dental records. PMID- 11490734 TI - [Basic principles of lacrimal apparatus surgery]. PMID- 11490735 TI - [Classical lacrimal apparatus surgery from the ophthalmological viewpoint]. PMID- 11490736 TI - [Endoscopic lacrimal duct surgery]. PMID- 11490737 TI - [Endonasal lacrimal apparatus surgery from ENT specialist viewpoint]. PMID- 11490738 TI - [Intracorneal ring segments for correction of myopia. Results of microsurgery technique]. AB - BACKGROUND AND OBJECTIVE: Since May 1998, small incision instruments for the implantation of intrastromal corneal ring segments (ICRS) are available enabling incisions only 1.2 mm in width instead of 1.8 mm to be made. After a follow-up period of 22 months in the conventional group and 8 months in the small incision group, the results of both groups were compared. PATIENTS AND METHODS: Since April 1997, 73 intrastromal corneal ring segments have been implanted, the first 43 ICRS implantations in 29 patients were performed using the conventional technique, the next 30 ICRS implantations in 22 patients were performed with the small incision technique. The average preoperative myopia was -3.5 D. RESULTS: After 6 months, 66.7% of the patients of both groups achieved the refraction target +/- 0.5 D. Even on the first postoperative day, the final refraction is achieved and shows stability for 1 year without regression. After 6 months, 80% of the conventionally operated patients and 77.8% of the patients operated on with the small incision technique showed an uncorrected visual acuity of 1.0 or better. With the small incision technique the preparation of the corneal dissection channel in 2/3 corneal stromal depth is significantly easier and can be carried out more safely. Perforations towards the inner or outer surface of the cornea can be practically excluded. Due to the smaller superficial wound in contrast to the conventional technique, a corneal suture is not necessary. Stromal depth of implantation was more than 50% and in both groups no decentration of 1.0 mm or more occurred. CONCLUSIONS: The small incision technique increases the intraoperative safety in the dissection of the stromal lamellae in the correct level. Both techniques are comparable with regards to visual acuity and refraction. The smaller corneal wound does not need a corneal suture. Because of these advantages we have completely switched our technique to the small incisions. PMID- 11490739 TI - [Optimizing laser parameters for intrastromal incision with ultra-short laser pulses]. AB - BACKGROUND: With the assistance of ultrashort laser pulses (ca. 200 fs pulse duration) it is possible to perform precise incisions inside the corneal stroma with a width of a few microns. The advantage of ultrashort pulses is that the required energy of a few microjoules is more than an order of magnitude lower compared with longer pulse durations, i.e. ps or ns pulses. Therefore, the secondary effects, such as thermal and mechanical damage to the surrounding tissue and the amount of radiation reaching the retina, are reduced. This method of intrastromal photodisruption allows a very well defined deposition of energy within the laser focus inside the corneal stroma, accompanied by minimal collateral damage. METHODS: The possibilities of performing intrastromal cuts using fs-laser pulses at a wavelength of 780 nm and pulse durations of 200 fs were studied using a titanium-sapphire laser system. The treated tissue samples were analysed by light and scanning electron microscopy to determine incision quality, reproducibility and achievable accuracy. The mechanical side effects of fs-photodisruption inside the surrounding tissue were analysed by pressure measurements using pyroelectric transducers. CONCLUSION: The thermal and mechanical side-effects of this method are very low and comparable to the effects during excimer treatment. Therefore an application of ultrashort laser pulses in refractive surgery appears to be a feasible alternative. PMID- 11490740 TI - [Follow-up of trabeculectomy. Complications and therapeutic consequences]. AB - BACKGROUND: Intensive follow-up after trabeculectomy is important for long-term success. In the postoperative period, complications must be detected early in order to initiate adequate therapeutic measures. PATIENTS AND METHODS: In a retrospective study 113 eyes from 113 consecutive patients were followed-up for a period of 6 months after trabeculectomy with regard to postoperative complications and therapeutic decisions (invasive and conservative). Criteria of success were defined as an intraocular pressure (IOP) < 21 mmHg without antiglaucomatous treatment as well as an IOP reduction > 20% compared to the preoperatively treated IOP. RESULTS: Most of the postoperative complications occurred within the first 3 weeks: scarring of the filtering bleb (47.6%), choroidal detachment (18.6%), shallowing of the anterior chamber (8%), fibrinous reaction (5.3%), overfiltration (4.4%), reduced flow under the scleral flap (4.4%), corneal dellen (3.5%), prolapse of the iris (3.5%), external fistulation (2.7%), increase of the IOP due to steroids (1.8%), detachment of the ciliary body (1.8%) and malignant glaucoma (0.9%). In the postoperative period, 45.1% of the eyes needed further interventions such as 5-fluorouracil (5-FU) injections (28.3%), needling and 5-FU-injections (14.2%), laser suturolysis (6.2%). Repositioning of the iris, YAG-laser iridotomy, laser treatment of the filtering bleb, autologous blood injection and cyclocryoapplication were needed only in rare cases and 36.3% received other conservative treatment (i.e. modification of topical steroids, increase of cycloplegic treatment). After 6 months the success rate without antiglaucoma medication was 88.5%. CONCLUSIONS: An intensive follow up after trabeculectomy is mandatory to avoid postoperative failure. In about half of the patients postoperative additional therapeutic measures became necessary, particularly if scarring of the filtering bleb occurred. Paying attention to these factors yields in a high success rate after filtering surgery. PMID- 11490741 TI - [Results of implantation of a Worst Iris Claw Lens for correction of high myopia]. AB - BACKGROUND: The purpose of this clinical study was to evaluate refraction, best visual acuity and anterior chamber flare after implantation of a Worst Iris Claw Lens to correct high myopia. PATIENTS AND METHODS: From September 1996 to February 1999 the implantation of a Worst Iris Claw lens was performed on 44 highly myopic phakic eyes of 28 patients. Data after 6 weeks and 6 months follow up were assessed. RESULTS: The mean preoperative spherical equivalent was--17.35 +/- 3.57 dpt and 6 months after implantation 79% of the eyes were within +/- 1.0 dpt of target refraction. The mean best corrected visual acuity increased from 0.43 preoperatively to 0.65 postoperatively and 6 months after the treatment there was no flare (< 10 photons/ms) in 21% and a mild anterior chamber flare (10 80 photons/ms) in 76% of the treated eyes. The median laser flare was 20 p/ms. CONCLUSIONS: In conclusion, implantation of a Worst Iris Claw lens has proved to be a precise method to correct high myopia. No or mild anterior chamber flare and a good visual acuity were measured in most eyes 6 weeks and 6 months after surgery. Longer follow-up periods are needed to assess the long-term safety and efficiency. PMID- 11490742 TI - [Morphological results after eccentric perforating keratoplasty]. AB - OBJECTIVE: Due to unfavourable optical and immunological conditions, an eccentric position of the corneal transplant is limited to curative or tectonic indications. The purpose of this study was to assess the morphological results of eccentric homologous penetrating keratoplasty (PK) with respect to diagnosis as well as position and size of the corneal graft. PATIENTS AND METHODS: In our retrospective study, 122 cases of eccentric homologous PK in 96 eyes between 01/1989 and 10/1997 were included, which in 104 patients was necessary because of a corneal ulcer (36 previous PK, 21 areactive/rheumatic, 20 herpetic, 15 bacterial, 12 other). Elective PK was performed on 6 eyes with corneal scars, 7 eyes with ectatic corneal diseases (ECT) and 5 others. Using postoperative slides the following parameters were quantified: the graft diameter (GD) and transparency, the distance of the geometric centre of the cornea from the central trephination margin (ABS) as well as from the geometric centre of the graft (decentration, DEZ). Of these, 17 grafts were classified as eccentric peripheral keratoplasty (EPK) with the optical axis through the host cornea and 67 as eccentric central keratoplasty (ECK) with the optical axis through the graft. RESULTS: The average follow-up period was 25 months. In EPK (ulcers only, 5.0 +/- 1.7 mm) the mean GD was significantly lower than in ECK (78% ulcers, 7.0 +/- 1.3 mm) (p < 0.001). The decentration of EPK (3.5 +/- 1.1 mm) was significantly higher than for ECK (1.4 +/- 0.8 mm, p < 0.001). ABS of EPK was +1.3 +/- 0.9 mm and for ECK was -2.0 +/- 1.0 mm. Of the grafts in EPK and ECK 46%/57%, respectively were crystal clear, 18%/15% respectively showed minor decompensation and 36%/28% were completely cloudy. In cases of ulcers/scars/ECT/others, 45%/75%/100%/75% were crystal clear, respectively, 18%/25%/0%/0% showed minor decompensation and 37%/0%/0%/25% were completely cloudy. With increasing stages of cloudiness, the mean graft decentration increased from 1.4 +/- 1.1 mm (crystal clear) to 1.7 +/- 1.2 mm (minor decompensation) and 2.2 +/- 1.4 mm (completely cloudy). Transplants with greater GD (p = 0.04) and ABS (p = 0.01) were significantly more often clear. Ulcers were significantly more cloudy than scars/ECT (p < 0.01). The position of the graft and its cloudiness was not significantly correlated. CONCLUSION: The diagnosis leading to eccentric PK seems to have the greatest impact on long-term graft transparency. Our results indicate that an increasing decentration of the graft is not inevitably associated with an increasing probability of clouding. A central penetrating repeat keratoplasty may be considered in the non-inflamed interval after eccentric tectonic mini keratoplasty. PMID- 11490743 TI - [Immunosuppressive therapy with mycophenolate mofetil (CellCept) in treatment of uveitis]. AB - BACKGROUND: Severe forms of uveitis can often only be managed sufficiently with systemic immunosuppression. All available drugs are known for their relative high rate of side-effects. Mycophenolate mofetil (MMF), an immunosuppressant successfully used in management after organ transplantation and many autoimmune diseases, has shown remarkably less side-effects when used for various forms of uveitis in monotherapy or in combination with corticosteroids. The aim of this multicenter-study was to investigate if monotherapy with MMF is effective in various forms of uveitis. METHOD AND PATIENTS: Ten patients with anterior uveitis (n = 3), intermediate uveitis (n = 2), panuveitis (n = 4) and retinal vasculitis (n = 1) were treated in a prospective study with 2 x 1 g MMF daily. Previous immunosuppression had been discontinued because of side-effects or ineffectivity in all patients. In these patients MMF was given in addition to the other immunosuppressant at the beginning of treatment. RESULTS: The follow-up time ranged from 1 to 12 months (mean 4.5 months). Under therapy with MMF (monotherapy in 4 patients, additional prednisolone in 5 patients and additional metotrexate in 1 patient) 8 patients remained free of recurrences. In one female patient depression of inflammation activity was only achieved after cessation of therapy with Cyclosporin A in combination with MMF and a switch to methotrexate. Another patient with a bilateral uveitis was free of recurrences in only one eye, the second eye did not develop recurrence due to the additional corticosteroid treatment. Side-effects were diarrhoea in one patient and probably gastrointestinal problems in another (leading to cessation of therapy in both patients) and in another case nausea, vomitus and alopecia 10 months after beginning therapy. CONCLUSIONS: MMF as a new immunosuppressant stopped inflammation or drastically reduced the rate of recurrences in 8 out of 10 patients with uveitis which was previously not brought under control by other immunosuppressants. The side-effects were tolerable in comparison with other immunosuppressive agents. More patients, longer follow-up times and a comparative study with Cyclosporin A are required to assess the long-term therapeutical success. PMID- 11490744 TI - [Ultrasound biomicroscopy diagnosis of traumatic choroid effusion without cyclodialysis]. AB - BACKGROUND: Chamber angle changes due to trauma represent a diagnostic challenge in modern ophthalmology and two examination techniques are compared: gonioscopy which has been used in ophthalmology for almost a century and is still undergoing continuous improvements and ultrasound biomicroscopy (UBM) which was introduced into clinical ophthalmology in 1991. CASE REPORT: We report the case of a 14-year old boy with ocular trauma caused by a soft gun projectile. Gonioscopy showed a large goniosynechia in the presence of ocular hypotension, therefore, cyclodialysis was suspected. However, a control investigation using UBM showed an intact and circularly attached but anteverted ciliary body behind the synechia, a circular choroidal effusion and an anterior displacement of the iris-lens diaphragm. CONCLUSION: In ocular trauma, UBM may under certain conditions clearly be of a higher diagnostic value than gonioscopy. Therefore, UBM should not only be considered as an additional examination technique in the evaluation of traumatic ocular pathologies but rather as the technique of choice. PMID- 11490745 TI - [Differentiation between dyslexia and ocular causes of reading disorders]. AB - BACKGROUND: Dyslexia is defined as a reading and/or writing disability persisting after exclusion of organic causes. Studies show that ocular disorders, especially small refraction errors, hypoaccommodation and symptomatic heterophoria, are often not detected or treated in cases of reading and/or writing problems which were otherwise diagnosed as dyslexia. METHODS: We evaluated the data of patients referred to our department from December 1997 to March 2000 with the diagnosis of dyslexia. RESULTS: We found ocular disturbances in 28 (84.8%) out of 33 children, 26 (78.8%) showed improved reading after therapy. They had mostly accommodative problems: uncorrected hyperopia, hypoaccommodation and/or exophoria compensated by accommodative convergence (pathophoria). CONCLUSIONS: Our results underline the importance of the correction of even small refraction and/or motility errors in the presence of reading and writing difficulties. PMID- 11490746 TI - [Spontaneous course of classical, subfoveal choroid neovascularization in age related macular degeneration]. AB - In a prospective, double-blind study, 19 patients with a classical subfoveal choroidal neovascularisation due to AMD were followed-up over a period of 2 years. Every 3 months a standardised visual acuity test, a contrast sensitivity test and fluorescein-angiography were performed. Overall, visual acuity dropped from 20/125 initially down to 20/250 after 2 years. Contrast sensitivity decreased from 23 down to 19 recognised letters. The functionally superior group 1 (n = 7) with a visual acuity of 20/65 at baseline, improved up to 20/50 after 2 years, while group 2 (n = 12) with a baseline visual acuity of 20/125 showed a decrease down to 20/400 after 24 months. Contrast sensitivity stabilised or improved in 10 patients from 22 to 25 letters, while in 9 patients the score dropped from 23 down to 8 recognised letters. The results underline the unfavourable functional prognosis of classical subfoveal lesions in AMD. However, stabilisation is possible depending on baseline visual acuity which is critical whenever results of nonplacebo-controlled studies have to be assessed. Moreover these results suggest that the urgency of a therapeutical intervention might be overestimated. As treatment, a fairly gentle, primarily atraumatic, CNV-selective strategy should be chosen, so that the self-limiting potential of this disease can be utilised in addition to the therapeutic effect. PMID- 11490747 TI - [Severe, bilateral vision loss in malignant melanoma of the skin. Anterior ischemic optic neuropathy with irreversible vision and visual field loss in adjuvant interferon alfa-2b therapy]. PMID- 11490748 TI - [Circular corneal opacities]. PMID- 11490749 TI - [Is diabetes in the elderly patient a risk factor for cataracts?]. AB - The higher prevalence of diabetes for cataracts as a component of the multifactorial genesis of age-related cataracts has been proven in several epidemiological studies. Most of these results identified diabetes as a highly significant risk factor for cataracts in older diabetic males and females. The increased cataract risk of diabetics then decreases after the age of 65 years and is related to different types of opacity. In addition to the sorbitol pathway, diabetic hyperglycaemia causes increased formation of glucose-derived advanced glycation cytotoxic end-products. In connection with these specific pathological changes, the crucial role of the lens epithelium with respect to cataractogenesis is emphasised in several studies. The results of our own prospective clinical study proved the significantly lower mean cell density in type II diabetics compared with eyes from non-diabetics. Together with the other determined morphological cell characteristics, this result seems to be due to the cataractogenic influence of diabetic metabolic disorders on the lens There are some indications for the primary cataractogenic importance of the lens epithelium in type II diabetes. PMID- 11490750 TI - [Searching for a position on the internet]. PMID- 11490751 TI - [Extreme challenges in hepatobiliary surgery]. PMID- 11490752 TI - [Adjuvant chemotherapy in liver malignancy for improving operability]. AB - Complete tumor resection is the only curative option for patients with colorectal liver metastases. Hepatic resection is frequently not possible for technical reasons: because of large tumors, multiple or bilateral metastases, or tumors that are too close to vessels. In these cases chemotherapy might downstage the tumor volume and facilitate secondary curative resection in patients initially not eligible for curative surgery. Treatment with fluorouracil (5-Fu) alone has resulted in disappointing response rates of about 10-20% in patients with colorectal liver metastases, which make these protocols useless in the neoadjuvant setting. Because regional chemotherapy into the hepatic arteria results in significantly higher response rates (40-50%), some studies have documented some success in secondary curative surgery after regional chemotherapy of initially unresectable colorectal liver metastases. However, regional chemotherapy is invasive and therefore not standard therapy for every patient with colorectal liver metastases. Recently new exciting treatment options have become available for colorectal cancer. Combinations of chemotherapy consisting of irinotecan and 5-Fu/FA or oxaliplatin and 5-Fu/FA result in response rates of 50% and can be considered a new standard first-line chemotherapy for patients with metastatic colorectal cancer. Recently, two encouraging retrospective studies have been published with chronomodulated chemotherapy of oxaliplatin and 5-Fu/FA in the setting of neoadjuvant chemotherapy for patients with unresectable colorectal liver metastases. With this multidisciplinary approach, antitumor activity of chemotherapy appears to be translated into a long-term survival benefit and some patients with initially unresectable colorectal liver metastases can potentially be cured. As a consequence, on the premises of close cooperation between surgeons and internists, more patients with metastatic colorectal cancer will be cured in the future. PMID- 11490753 TI - [Multistage liver resections in colorectal liver metastases. The Paul Brousse concept]. AB - Hepatic resection is currently the only form of treatment that offers a chance of long-term survival, with rates ranging from 25% to 39%. However, a curative operation can be performed in only 10% of patients with colorectal metastases to the liver. Our policy is to increase the number of patients that can benefit from liver resection. Liver metastases can be considered as irresectable mainly in three different situations (sometimes associated): (I) large and/or poorly located tumors; (II) bilateral tumors in both liver lobes; (III) tumors technically resectable, but not operable because the liver remnant is too small, which is associated with a prohibitive risk of postoperative severe liver failure. The aim of this paper is to report the strategies we use in our center to achieve curative resection in these three schematic situations despite initial contraindications. PMID- 11490754 TI - [Preoperative portal embolization for induction of liver hypertrophy]. AB - Preoperative portal embolization (PVE) is now widely used to induce hypertrophy of the anticipated liver remnant. Here, pathophysiology, indications, technical procedures, and problems of PVE are reviewed and our experience of 105 PVEs is presented. The reported complication rate after PVE is low (0-10%) and the increment of the future remnant liver volume is 12% of total liver volume (range: 7-27%). The resection rate after PVE ranged from 58% to 100%. In our series of 105 PVEs, the morbidity rate was 3.8%, the future remnant liver increased by about 10% of total liver volume, and 34 patients (32.4%) did not undergo planned extended hepatectomy. After hepatectomy, hepatic failure occurred in one patient (< 1%). In conclusion, although PVE is a safe procedure for enlarging the future remnant liver, more appropriate indication criteria should be established. PMID- 11490755 TI - [Surgical therapy of liver hilus tumors]. AB - Hilar resections and hemihepatectomies as surgical strategies for hilar cholangiocarcinoma achieve only limited rates of resectability and radicality. Principles of surgical oncology have to be applied in order to increase the numbers of patients undergoing resection as well as their long-term survival. Due to the anatomical architecture of the hepatic hilum and side-specific variations within the biliary tree, right trisectorectomy and principal portal vein resection have the potential to comply with basic rules of surgical oncology, i.e. wide tumor-free margins and a no-touch dissection technique. In our experience, 5-year survival after formally curative right trisectorectomy and portal vein resection is 65% in spite of advanced tumor stages. Resection of the entire biliary tract without dissection of the tumor is possible by combining total hepatectomy, partial pancreatoduodenectomy and liver transplantation. However, even this procedure does still not fully prevent tumor cell dissemination. So far, a putative alteration of tumor cell kinetics due to posttransplant immunosuppressive treatment results in an increased rate of implantation metastases. PMID- 11490756 TI - [Liver resection in liver cirrhosis]. AB - The benefits of liver resection for patients with hepatocellular carcinoma (HCC) and concomitant liver cirrhosis have been questionable due to high rates of postoperative complications. As a result of advance in surgical techniques, along with improved perioperative management and increased knowledge of the pathophysiology of liver cirrhosis, liver resection in cirrhotic patients has become a safer surgical procedure during the 1990s. This article introduces our techniques of parenchymal resection in patients with liver cirrhosis, avoiding inflow occlusion of the hepatic circulation by using a Cavitron Ultrasonic Surgical Aspirator (CUSA) and bipolar cautery that is equipped with a mechanism for channeling water at the point of cauterization. Analysis of survival rates showed that surgical resection was more advantageous than treatment such as percutaneous ethanol injection, although the discovery of multicentric carcinogenesis in livers with hepatitis C viral infection required us to reconsider surgical indications for HCC and concomitant liver cirrhosis. PMID- 11490757 TI - [Definition of surgical degree of freedom by functional anatomy in liver resection surgery]. AB - Liver resections have developed to very complex and differentiated operations, clearly adapted to individual anatomical and physiological conditions. In parallel, perioperative morbidity has been dramatically reduced. Intraoperative strict consideration of various details of hepatic anatomy, particularly of functional liver anatomy, has proved to be of particular importance when liver surgery reaches indication and technical limits. The term "functional anatomy" stands for a form of hepatic substructurization, which is primarily based on the existence of hemodynamically independent regions of liver parenchyma. A selection of some of the most important details and facts of functional liver anatomy and secondary derived guidelines for surgical strategy and technique is presented in an overview, with special focus on liver resection. PMID- 11490758 TI - [Pancreatoblastoma: a rare malignant neoplasm in early adulthood]. AB - INTRODUCTION: Here we report the ninth case of pancreatoblastoma in an adult. PATIENT AND METHOD: At the time of diagnosis the female patient was 22 years old. She had a tumor in the left upper abdomen, measuring 9 cm in diameter. Preoperatively the diagnosis of a pancreatoblastoma was suspected and an explorative laparotomy was performed followed by radical tumor extirpation and intraoperative radiotherapy. After the diagnosis of a pancreatoblastoma was confirmed by definitive histopathological examination, combined Radiochemotherapy was performed. DISCUSSION: Special features in histomorphological characterization of the tumor and problems concerning diagnosis and differential diagnosis as well as the lack of a definitive therapy regime are discussed. PMID- 11490759 TI - [Selective pre- intra- and postoperative bile duct diagnosis--an efficient and low complication regimen within the scope of laparoscopic cholecystectomy. Early and late results of a prospective study]. AB - INTRODUCTION: The discussion of diagnostic pre- and intraoperative procedures of the common bile duct (CBD) for laparoscopic cholecystectomy is still open. METHODS: Results of a prospective study of 4000 laparoscopic cholecystectomies were evaluated, adhering to the concept of selective common bile duct examination. In 1999 an inquiry was performed on 1197 patients an average of 38.7 months postoperatively to judge this concept. RESULTS: Three hundred and nineteen patients (8%) examined preoperatively by ERC revealing a CBD stone in 126 cases. The morbidity of this procedure was 3.1%. In 0.18% a lesion of the common bile duct was documented following laparoscopic cholecysTectomy. A intraoperative cholangiography was performed in 67 patients (1.7%) because of unclear anatomy or to exclude a CBD stone. Perioperatively a CBD stone was found in 0.5% of cases. Additionally, in 0.3% the same problem occurred in the long-term follow-up, so that the risk for unknown CBD stones following the selective diagnostic concept is 0.8%. CONCLUSION: The concept of selective diagnostic procedures of the common bile duct shows a low morbidity with sufficient efficiency in avoiding CBD stones as well as duct lesions. Therefore it is recommended for routine use in laparoscopic cholecystectomy. PMID- 11490760 TI - [PET examinations of recurrent rectal carcinoma, 2]. AB - We have shown previously that the bodies of vertebrates are made of a somatic and a visceral component, separated by thin fascial sheets (Chirurg 66: 1230). We postulated that advanced visceral cancers are confined in their pattern of lymphatic spread to one or a few distinct metastatic pathways. In this they are fundamentally different from somatic cancers. To investigate this hypothesis in vivo, we evaluated positive lymph nodes (LNs) on PET scan series of 58 consecutive patients with rectum cancers (RC) compared to breast cancers (BC). We conclude that in RC, a visceral cancer spreads mostly along only one caudocranial pathway (regional, para-aortic, mediastinal, supraclavicular LNs) and never affects somatic LNs. In contrast, BC and urogenital carcinomas, as somatic cancers spread to regional somatic LNs, but do not affect visceral LNs. PMID- 11490762 TI - [Jejunum interposition as replacement of the pars descendens duodeni in extensive tubulovillous adenoma]. AB - Reconstruction after partial duodenectomy with resection of the ampulla of Vater is often troublesome. We report the case of a 70-year-old patient with endoscopically non-resectable tubulo-villous adenoma of the descending duodenum including the ampulla of Vater in which subsequent biopsies revealed dysplastic areas. A partial resection of the descending duodenum including the ampulla of Vater was performed. Reconstruction was achieved by the interposition of a jejunal limb in which the ampulla could be reinserted to the posterior wall. The postoperative course was uneventful; a carcinoma was not found within the specimen. In cases of widespread adenomas of the ampulla of Vater, duodenum preserving resection by interposition of a jejunal limb with reinsertion of the ampulla into the posterior wall may be used as an alternative to Roux-Y reconstruction and to Whipple's procedure. PMID- 11490761 TI - [Long-term outcome of stomach carcinoma achieved in an Austrian standard hospital with an oncologic focus]. AB - INTRODUCTION: Although two large prospective and randomized planned European studies failed to show any benefit of radical D2 lymphadenectomy for gastric cancer, the value of radical lymphadenectomy is still a matter of controversy. METHODS: A radical surgical approach principally using D2, D3 lymphadenectomy, as defined by the Japanese Research Society for Gastric Cancer, has been prospectively performed since January 1984. Out of 729 patients with gastric cancer, 521 were surgically treated for potential cure between 1984 and 31 December, 1998. Clinical, histopathological and surgical factors were evaluated for their influence on long-term survival by means of univariate and multivariate analysis. RESULTS: Tumor-specific 5- and 10-year survival rates for all patients were 58.5% and 57.5% for patients who underwent tumor resection 59% and 58%. For operated patients upon with the aim of achieving cures, the tumor-specific 5- and 10-year survival rates were 63.3% and 62.2% and the median survival time was more than 144 months. Postoperative hospital mortality was 7.7%, 4.6% for R0 resected patients, 8.6% for R1,2 resected patients and 21.3% for those undergoing palliative procedures. Multivariate analysis using the Cox model identified an age older than 65, total gastrectomy as well as high pN- and pT category as detrimental factors with an independent influence on survival. CONCLUSION: After updating the long-term results of gastric cancer, as already published earlier, it is impressively obvious that also in a European setting of gastric cancer patients, with a presupposed appropriate surgical technique and experience, very constant cure rates are achievable with comparatively low mortality and morbidity. PMID- 11490763 TI - [Blunt trocar with modified tip for open laparoscopy]. AB - A blunt trocar for open laparoscopy is presented. Its modified design leads to better visual control of the trocar tip during insertion. Thus the risk of misplacement of the trocar is reduced. PMID- 11490764 TI - [Intra-gastric penetration of an adjustable gastric band]. AB - Between November 1995 and August 2000 we performed adjustable silicone gastric banding laparoscopically in 252 patients. The body mass index varied from 37 to 86 kg/m2. We report on a 38-year-old woman who was operated on in 1997 with a body mass index of 47 kg/m2 (167 cm, 132 kg). The postoperative follow-up was uneventful until January 2000. The patient lost weight until she weighed 78 kg. Then she complained of diffuse epigastric pain. Gastroscopy revealed gastritis. Omeprazol was prescribed. No amelioration occurred. Endoscopic control showed partial intragastric migration of the band. After laparoscopic removal of the band, the patient was free of symptoms. Band erosion is a possible complication of adjustable gastric banding. As is known from intragastric penetration of the Angelchik prosthesis, the clinical symptoms of this complication may be mild. Since the follow-up of most patients with gastric banding is less than 5 years, more complications similar to that one described may be diagnosed in the future. PMID- 11490765 TI - [Leiomyosarcoma of the splenic artery]. AB - Leiomyosarcomas of vascular origin are a very rare group of tumors developing from smooth vascular muscle cells. The most common location are in large vessels, e.g. vena cava, saphenous vein or pulmonary artery, but fewer than 200 cases are reported in the literature. We report a case of a patient with a leiomyosarcoma of the splenic artery. This location has not been described previously. PMID- 11490766 TI - [Fibrovascular esophageal polyp--diagnosis and therapy]. AB - Fibrovascular polyps are extremely rare benign neoplasms of the esophagus. The most prominent clinical symptom is enoral tumor regurgitation, which may lead to asphyxiation by pharyngeal impaction. Usually fibrovascular polyps cause dysphagia and progressive weight loss. Diagnosis by endoscopy and barium swallow may be unexpectedly difficult. The most frequent incorrect diagnoses are achalasia or an intramural or mediastinal tumorous mass compressing the esophagus. CT scan and MR imaging are of little help. Small polyps may be resected endoscopically by means of electrocautery or Nd:YAG laser ablation. In most cases, however, surgical resection is required. Since the basis of the polyp is usually located subcricoidally, tumor exposure and resection are achieved by esophagotomy via a left cervical approach. Thoracotomy is seldom required. PMID- 11490767 TI - [Development of modern physical diagnosis and its application in cardiothoracic surgery]. AB - The year 2000 marked the 300th anniversary of the birth of Gerard Van Swieten (1700-1772). He reformed medicine in Vienna, putting new emphasis on diagnosis based on clinical observation in combination with anatomical-pathological findings. This led to the introduction by Leopold Auenbrugger (1722-1809) of percussion in the examination of thoracic organs. However, further work by Joseph Skoda (1805-1881) was required to finally establish this method as a diagnostic tool for pathologies in the thorax. The surgeon Franz Schuh (1804-1865) carried out further basic research in respiratory physiology that cleared the way for the use of percussion and auscultation in thoracic surgery. He is also remembered for introducing experimental surgery in Austria, thus making surgery a science. This article aims to recall these men and their fundamental work behind early paracentesis of the thorax, and especially the first successful pericardiotomy by Schuh 160 years ago, which is also considered a milestone in cardiac surgery. PMID- 11490768 TI - [Injuries of the thoracic and lumbar spine]. PMID- 11490769 TI - [Organizational consequences for introduction of diagnostic related groups (DRGs)]. PMID- 11490770 TI - [Agreement on elective procedures in unconscious (emergency) patients]. PMID- 11490771 TI - [Business tax responsibility for medical expert assessment]. PMID- 11490772 TI - [Legal and organizational possibilities of assistant change from the viewpoint of hospital administration]. PMID- 11490773 TI - [Differential diagnostic problems in assessment of benign bone tumors]. AB - The diagnosis of benign bone tumors can cause many problems and a definitive histological diagnosis is often impossible without the knowledge of clinical datas, age of the patient and anatomical location of the tumor. Therefor cooperation between pathologist, orthopedic surgeon and radiologist is necessary, to reduce the risk of misinterpretation of benign and malignant bone tumors. Benign bone tumors often show a heterogenous histology and overlapping morphology between different entities has to be kept in mind, as well as the sampling error in biopsy- and frozen section material. In most cases the pathologist has to rely on conventional stainings because the immunohistochemistry stainings are often not helpful. PMID- 11490774 TI - [Benign bone-forming tumors]. AB - Benign bone-forming tumors include osteomas, enostomas, osteoid osteomas, and osteoblastomas. These lesions are often characterized by typical imaging findings on radiographs, CT and MR imaging studies. Radiologic findings and additional clinical information allow for a specific diagnosis in most cases. This review article emphasizes the radiological patterns of benign bone-forming tumors as well as their epidemiological, clinical, and pathological characteristics. In addition, minimally invasive interventional procedures for the therapy of osteoid osteoma are reviewed. PMID- 11490775 TI - [Benign cartilaginous tumors]. AB - Benign cartilaginous tumors are the most frequent bone tumors. At radiography they are not seldomly detected as an additional finding. Most osteochondromas and enchondromas can be diagnosed radiologically based upon their morphology. However, differentiation of a low grade chondrosarcoma from an enchondroma is difficult, and differentiation of a peripheral chondrosarcoma from an osteochondroma can be difficult. Clinics, localization, age distribution and morphology of benign cartilaginous tumors are presented and the most important differential diagnoses are given. PMID- 11490776 TI - [Cysts and cystoid formations]. AB - Cystic and cyst-like lesions of bone are very common. They can be of primary origin or secondarily caused by several diseases. For exact diagnosis or at least narrow differential diagnosis the assessment of radiological findings is important. However, patient data, clinical symptoms, laboratory parameters, and site of the lesion have to be included for a reasonable final diagnosis or differential diagnosis. PMID- 11490777 TI - [Diagnostic imaging of giant cell tumors]. AB - Although classified as benign neoplasms, giant cell tumors are locally aggressive with a high recurrence rate of 30-50%. The histological appearance include osteoclast-like, multinucleated giant cells and round to spindle-shaped mono nuclear stromal cells. These neoplasms predominately affect adults in the third and forth decades of life (70-80%). Giant cell tumors predominately arise in long tubular bones (75-95%) with the majority occurring around the knee (50%). The next most common site being the distal radius (10%). The epicenter of giant cell tumors is in the epiphysis. The radiographic signs of giant cell tumors are a geographical radiolucency with no internal mineralization, a thinning of the cortex, eccentric in the epiphysis of long bones. MRI signs of giant cell tumors are high signal intensity in T2-weighted images, high contrast media enhancement, fluid levels, signs according to haemorrhage and haemosiderin deposition. CT shows the expanded and thinned cortex. Plain radiographs remain the mainstay of diagnosis of giant cell tumors. MRI and CT are important for staging and therefore for surgical planning. PMID- 11490778 TI - [Characteristic perfusion pattern of osseous giant cell tumor in dynamic contrast enhanced MRI]. AB - PURPOSE: To describe the perfusion pattern of giant cell tumor (GCT) of bone with Gd-enhanced dynamic MR imaging. To compare time-intensity-curves in patients with local recurrence and postoperative alterations without recurrence. METHODS: Nine patients (5 women, 4 men) with GCT of bone underwent 19 dynamic MRI examinations. Mean age was 34 years (range 24-64 years). All diagnoses were proven by pathology. Dynamic contrast-enhanced MRI was performed at 1.0 T using T1-weighted gradient echo sequences. GCT was located in the distal radius (4x), tibia (3x), fibula (1x) and humeral head (1x). RESULTS: All giant cell tumors showed a uniform perfusion pattern with a steep slope and maximum intensity value followed by an early and rapid washout phase. The same pattern appeared in five local recurrences of GCT in four patients. In nine follow-up examinations without local recurrence dynamic MRI yielded in uncharacteristic perfusion patterns. CONCLUSION: These results demonstrate a uniform perfusion pattern of GCT of bone obtained by dynamic MRI. It is characterized by a steep slope followed by an early and rapid washout phase. This characteristic pattern can also be obtained in local recurrences. Dynamic contrast-enhanced MRI appears a helpful method for primary diagnosis of GCT of bone and detection of local recurrences after surgery. PMID- 11490779 TI - [Diagnostic-therapeutic procedure in benign bone tumors]. AB - The indications of surgical treatment of benign bone tumours are strongly related to the clinical behaviour, the patient's complaints and the activity in Tc-bone scan. Some lesions--like the nonossifying fibroma--that can safely be diagnosed by conventional x-rays may not be treated surgically--as long as the patients are free of pain, and there is no risk of pathologic fracture. In case of clinically relevant lesions, activity in Tc-bone scan, or a risk of fracture, biopsy and curettage combined with autologous or homologous bone grafting is indicated. Some aggressive tumors, like the giant cell tumor, have to be treated more aggressively by curettage with adjuvant measures (Phenole) or by marginal excision. PMID- 11490780 TI - [Post-traumatic dissection of the abdominal aorta]. AB - Dissections due to deceleration trauma are rarely limited to the infradiaphragmal aorta (only 2-3%) and are usually lethal. Here we report the unusual course of an abdominal aortic dissection with aneurysmatic enlargement of the false lumen. Based on diagnostic imaging, a therapeutic stent application was planned in order to close the entry and to prevent rupture. During the intervention sondation of the false lumen revealed that the left renal artery had a reentry. Due to the complexity of the entry-reentry situation of the left renal artery the intervention was not possible, and the patient had to undergo vascular surgery. PMID- 11490781 TI - ["Service oasis USA". ...or experiences with a book order from an American publisher]. PMID- 11490782 TI - [Conspicuous MRI image of the spine after treatment of a medulloblastoma. Diffuse bone marrow metastasis of medulloblastoma]. PMID- 11490783 TI - [Cerebral ischemia]. PMID- 11490784 TI - Spontaneous biliary peritonitis in acalculous cholecystitis: fact or misdiagnosis? AB - It is often speculated that an inflamed gallbladder weeps bile to produce bile peritonitis. This may be so, but more likely the problem is a peritoneal effusion in a jaundiced patient which thus resembles bile. So-called "spontaneous or idiopathic biliary peritonitis" in acute acalculous cholecystitis without a proven cause is a further example of this very rare condition. Spontaneous perforations of the extrahepatic biliary ductal system associated with acalculous cholecystitis are uncommon albeit reported in adults. Most patients present with an acute abdomen and are operated upon urgently without diagnostic iter. A recent experience with such a case prompted a thorough review of 27 similar cases previously reported. PMID- 11490785 TI - DIC-CT findings of biliary cystadenocarcinoma communicating with the bile duct: a case report. AB - Among the intrahepatic cystic diseases except Caroli's disease, only biliary cystadenoma/cystadenocarcinoma may communicate with the bile duct. We present a case of biliary cystadenocarcinoma in which drip infusion cholangiographic computed tomography demonstrated communication between an intrahepatic cyst and the biliary system preoperatively. Drip infusion cholangiographic-computed tomography, a simple and noninvasive examination, is useful for differentiating biliary cystadenoma/cystadenocarcinoma from other intrahepatic cystic lesions. PMID- 11490786 TI - Correlation between intestinal metaplasia of the gastric cardia and gastroesophageal reflux disease. AB - BACKGROUND/AIMS: IMC (intestinal metaplasia of the cardia) has been a subject of great interest, given the rapidly increasing incidence of adenocarcinoma in this location, over the past two decades. Whether this histological alteration is a consequence of gastroesophageal reflux disease, or a manifestation of an H. pylori-related multifocal atrophic gastritis, is unclear. Furthermore, whether IMC should be considered a premalignant lesion of gastric cardia is still unknown. We performed a prospective study in order to determine the prevalence of IMC in patients presenting for elective esophagogastric-duodenal endoscopy and to evaluate a potential association between IMC and some clinical, endoscopic and histological variables. METHODOLOGY: Biopsy specimens were taken from 105 unselected patients undergoing routine diagnostic endoscopy. Eight biopsies were taken from different sides, for histological evaluation: 1 above and 3 below the squamocolumnar junction, 2 from gastric fundus and 2 from gastric antrum. All specimens were stained with hematoxylin and eosin-Alcian blue and modified Giemsa to facilitate the detection of H. pylori. RESULTS: Eighty-six patients (50 males, 36 females) with a mean age of 46.5 years (range: 23-75 years), were included in the study. Twenty-one (24.5%) were found to have IMC; 4 (19%) of these had concomitant low-grade dysplasia of the cardiac mucosa. IMC was associated with: males (P = 0.04), endoscopic diagnosis of esophagitis (P = 0.02), histological diagnosis of esophagitis (P = 0.008), mucosa of the cardiac type (P = 0.02), chronic carditis (P = 0.002) and dysplasia (P = 0.04). There was no correlation with: age, reflux symptoms, activity of carditis, H. pylori infection of the cardia and intestinal metaplasia of the distal gastric mucosa. CONCLUSIONS: IMC is common in our area. It is associated with endoscopic and histological changes of gastroesophageal reflux disease, but not with H. pylori infection of the cardia. Although 19% of patients presented concomitant dysplasia (of low grade), long-term follow-up studies will be necessary to assess the effective risk of IMC for cancer transformation. PMID- 11490787 TI - The effect of augmenting portal venous inflow on intrahepatic pressure and resistance in the isolated perfused porcine liver. AB - BACKGROUND/AIMS: The literature regarding the relationship between portal venous flow and pressure is controversial. The aim of this study was to examine the effects of doubling portal venous inflow on hepatic hemodynamics. METHODOLOGY: Portal venous pressure, intrahepatic portal venous resistance, hepatic arterial pressure and intrahepatic arterial resistance were assessed during basal portal venous inflow (756 +/- 142 mL/min; mean +/- SD) and enhanced portal venous inflow (1512 +/- 284 mL/min) in an isolated perfused normal porcine liver model (n = 6). Hepatic arterial flow was maintained constant throughout the experiments. RESULTS: During the period of enhanced portal venous flow there was an increase in: portal venous pressure (from 9 +/- 2 to 22 +/- 7 mm Hg, P = 0.0076); the difference between portal venous and hepatic venous pressures (from 2 +/- 2 to 10 +/- 5 mm Hg; P = 0.0289); hepatic arterial pressure (from 84 +/- 9 to 151 +/- 33 mm Hg, P = 0.0019); and intrahepatic arterial resistance (from 0.3488 +/- 0.0637 to 0.6387 +/- 0.2020, P = 0.0046). CONCLUSIONS: The increases in hepatic artery pressure and intrahepatic arterial resistance are a result of the hepatic arterial 'buffer response', a phenomenon not previously demonstrated in vitro. The magnitude of the observed changes in portal venous and hepatic venous pressure leads to the conclusion that, in the porcine liver, the intrahepatic venous resistance sites react by constricting to increases in portal venous inflow. PMID- 11490788 TI - Study on endoscopic esophageal mucosal resection with ligating device. I- Clinical study. AB - BACKGROUND/AIMS: EEMRL (endoscopic esophageal mucosal resection with a ligating device) has become increasingly popular. In this article, we review 13 clinical cases of EEMRL. METHODOLOGY: Since 1993, we have performed EEMRL to treat 15 lesions in 13 patients. Twelve squamous cell carcinomas (mucosal cancer in 10 and submucosal cancer in 2) were included among the 15 lesions. RESULTS: EEMRL failed to achieve complete resection of the 2 submucosal lesions (3.0 and 2.8 cm in maximum diameter). However, esophageal lesions could be removed successfully when 2.5 cm or less in maximum diameter. The procedure was not associated with any complication. CONCLUSIONS: Our clinical study showed that this technique may be indicated for esophageal cancer with a maximum diameter < or = 2.5 cm and confined to the mucosa. EEMRL is a technically easy and minimally invasive therapy which could be useful for the treatment of early esophageal cancer. PMID- 11490789 TI - Study on endoscopic esophageal mucosal resection with ligating device. II- Experimental study. AB - BACKGROUND/AIMS: This study reports on animal experiments regarding the safety of endoscopic esophageal mucosal resection with a ligating device (EEMRL), as well as the amount of mucosa which can be removed by this technique, the depth of resection and the feasibility of piecemeal resection. METHODOLOGY: Three experiments were performed in six mongrel dogs under general anesthesia. RESULTS: When EEMRL was done without submucosal injection of saline, resection reached the muscular layer and caused esophageal perforation. The average dimensions of the mucosal pieces resected using 8-, 10-, and 12-mm devices was 13 x 10 mm, 18 x 15 mm, and 22 x 18 mm, respectively. Resection reached the mid-plane of the submucosa and the depth was almost uniform. After piecemeal resection, there was no macroscopically visible mucosa at the resection site and each mucosal piece was resected along the mid-plane of the submucosa. CONCLUSIONS: The experimental study indicated that submucosal injection of saline is essential to prevent esophageal perforation. It also showed that EEMRL allows resection up to the mid plane of the submucosa, that the 12-mm device allows en bloc resection of lesions < or = 15 mm in diameter and that EEMRL is suitable for piecemeal resection. PMID- 11490791 TI - Spontaneous perforation of the right hepatic duct. AB - Spontaneous perforation of the biliary ductal system is a rare cause of peritonitis with a high morbidity and mortality rate. We present a case of an 80 year-old female who presented to the Emergency Department with acute onset abdominal pain and peritoneal signs. She was taken to the operating room for exploratory laparotomy and discovered to have a perforation of her right hepatic duct. Several risk factors were found to play a role in her biliary perforation: biliary calculi, infection, and vessel thrombosis. She underwent a cholecystectomy, choledochotomy with removal of gallstones, repair of the perforation, and placement of a T-tube. She had a prolonged recovery in the intensive care unit and was eventually discharged to a skilled nursing facility. We conclude that the management of this unique and highly fatal disease can be applied with an excellent outcome. PMID- 11490790 TI - Splenectomy enhances liver regeneration through tumor necrosis factor (TNF)-alpha following dimethylnitrosamine-induced cirrhotic rat model. AB - BACKGROUND/AIMS: We demonstrated that partial splenic embolization for hematological disorders in cirrhotic patients also improved liver function. Therefore, we investigated the mechanism of the beneficial effects of splenectomy on a rat cirrhotic model. METHODOLOGY: 1) Rats were administered DMN (dimethylnitrosamine) after splenectomy (splenectomized DMN rats) or a sham operation (DMN rats). 2) After completion of DMN administration, a tumor necrosis factor-alpha inhibitor (E3330) was administered on the same day as the splenectomy. Histological examination and cytokine expressions were analyzed. RESULTS: The splenectomy apparently reduced liver damage. This may be partially due to the enhancement of liver regeneration since the proliferating cell nuclear antigen labeling index in the DMN-treated liver was significantly increased by splenectomy. Tumor necrosis factor-alpha was down-regulated in the DMN rats, whereas its expression was preserved in the splenectomized DMN rats. There were no apparent differences in the number of Kupffer cells between the splenectomized DMN and the DMN rats, suggesting that the down-regulation of tumor necrosis factor-alpha may contribute to the reduction of Kupffer cells' function. In addition, a tumor necrosis factor-alpha production inhibitor (E3330) significantly reduced the proliferating cell nuclear antigen labeling index after splenectomy. CONCLUSIONS: Splenectomy, in this model, may promote liver regeneration by preserving Kupffer cell function, especially the secretion of tumor necrosis factor-alpha. PMID- 11490792 TI - Untractable diarrhea due to late onset celiac disease of the adult following pancreatoduodenectomy. AB - This case describes a rare cause of severe diarrhea that may occasionally plague the postoperative course of pancreatic resections. Although exceedingly rare this complication has already been described in another two patients undergoing pancreatoduodenectomy with diagnosis of duodenojejunal adenocarcinoma. Surgeons dedicated to pancreatic resections as well as pancreatologists caring for these patients should retain this possibility in their cultural background. Diarrhea, due to either exocrine insufficiency or interruption of sympathetic nerves to the small intestine, is a common complaint after pancreatoduodenectomy. A 54-year-old white female after a pancreatoduodenectomy with standard lymphatic clearance developed intractable diarrhea leading to severe cachexia. Recognition of the underlying occult celiac disease and institution of the appropriate dietary regimen allowed quick and expeditious recovery. PMID- 11490793 TI - Primary squamous cell carcinoma of the stomach. Report of a case and review of literature. AB - Primary squamous cell carcinomas of the stomach represent a rare entity. Since the first report in 1895 by Rorig et al. (1) only 80 cases have been published. These reports show a peak incidence in the sixth decade of life and preference of male gender (5:1). We report the case of a 61-year-old patient who presented with anemia and weight loss due to a large tumor of the gastric wall with adhesion to the pancreatic tail. After radical regional "en bloc" gastrectomy, splenectomy and pancreatic tail resection, the diagnosis of primary gastric squamous cell carcinoma could be confirmed, since the esophageal wall and the pancreatic tail were not infiltrated and extragastric squamous cell primaries could be excluded. After postoperative irradiation of the upper abdominal area, the patient developed a single liver metastasis in the left hepatic lobe that decreased with polychemotherapy. It was resected half a year later. Due mainly to advanced tumor stages, survival after surgical resection is poor. However, adjuvant radio and chemotherapy have resulted in survival rates of more than 3 years in reported cases, as in the present case. Five years after the diagnosis was established the patient is free of recurrence and without any complaint. Pathophysiological features, therapy and outcome are discussed by reviewing the cases reported in world literature. PMID- 11490794 TI - Retroperitoneal localization of hydatid cyst disease. AB - In this study we discuss a rare case; hydatid cyst disease that is located in the retroperitoneum. Two cases were treated in our department with hydatid cyst disease located in the retroperitoneum; one of the cysts was in the minor pelvis and the other was on the left psoas muscle. Diagnosis was confirmed with blood tests, and radiological examinations. The cyst wall was excised partially and was oversewn. Following the operation both of the patients were given albendasole tablets for 10 weeks with blood count and liver enzyme monitorization. Hydatic cysts located in retroperitoneum were treated successfully. Especially in the endemic areas hydatid cyst should be remembered when evaluating cystic masses in the retroperitoneum. It can be treated successfully with surgery. PMID- 11490795 TI - Tumor immunity and immunotherapy. AB - Recent advances in understanding fundamental mechanisms of regulation of the immune response are throwing light into mechanisms of tumor growth. The understanding of the immunological aspects of tumor expansion is leading to the development of new strategies to stimulate the immune system to mount more effective responses to tumors. This progress as well as some promising results in clinical trials is generating considerable optimism. The goal of this review is to present recent advances concerning tumor immunology, identify some of the pitfalls and problems that still exist and present new areas of immunotherapy that look promising. PMID- 11490796 TI - Laparoscopic colectomy for sigmoid diverticulitis: a prospective study in the elderly. AB - BACKGROUND/AIMS: The aim of this prospective study was to determine the feasibility and the complications or benefits of laparoscopic colectomy for sigmoid diverticulitis in patients aged 75 years or more. METHODOLOGY: From January 1993 to December 1999, 85 patients underwent an elective colectomy for sigmoid diverticulitis. Twenty-two patients over 75-years old (group 1) were compared to 63 younger patients (group 2). RESULTS: In group 1, there were 12 women and 10 men, with a mean age of 77.2 years (range: 75-82); In group 2, there were 35 women and 28 men, with a mean age of 53.7 years (range: 38-74) (P = 1.10 14). The operative time was shorter in group 2 (183 vs. 234 min). There was no difference between the 2 groups with regard to the postoperative period during which parenteral analgesics were required (5.4 vs. 5.2 days, P = 0.48) and the postoperative morbidity (18% vs. 14%, P = 0.06). Postoperative length of hospital stay (13.1 vs. 8.8 days, P = 0.003) was shorter in group 2 than in group 1. There was no perioperative mortality. Conversion rate was 9% (group 1) and 6% (group 2) (P = 0.6). CONCLUSIONS: In summary, data from the present study suggest that laparoscopic colectomy for sigmoid diverticulitis can be applied safely to older patients with fewer complications, less pain, shorter hospital stay and a rapid return to preoperative activity levels. PMID- 11490797 TI - Successful treatment for the IVC syndrome due to recurrence of colon cancer- chemotherapy in combination with the use of the expandable metallic stent placement. AB - A 73-year-old woman developed severe edema of the abdominal walls and legs due to rapid regrowth of unresectable paraaortic lymph node metastasis from ascending colon cancer. The expandable metallic stent was placed at the site of stenosis. Following the stent placement, she had marked improvement in her urinary output, ascitic drainage and edema of the lower extremity. Subsequently, intravenous infusion of 5-fluorouracil with high-dose leucovorin was performed. The irritable symptom disappeared completely after 5 weeks, and the patient recovered the ability to walk. The patient is alive 22 months after primary noncurative operation. Symptoms did not recur 4 months after the placement of the stent. The authors recommend the multimodality for palliation therapy in unresectable colorectal cancer. PMID- 11490798 TI - Idiopathic chronic constipation: pathophysiology, diagnosis and treatment. AB - Constipation is one of the most common disease in Western countries. Its prevalence is widely variable (5-30%), depending on criteria utilized for diagnosis. Usually constipated patients have mild-moderate symptoms and are self treated. Only patients that do not benefit from simple therapy (dietary supplementation of fiber and water, laxatives) need examinations such as blood tests, colonoscopy, radiography with bario-enemas, intestinal transit time and manometry. These tests are able to differentiate idiopathic from secondary constipation. Idiopathic chronic constipation is a collective term that involves different functional disorders. More specific investigations (oro-cecal transit time, manometry, electromyography) allow one to distinguish between two different patterns of idiopathic chronic constipation (slow transit constipation and pelvic floor disease) and to utilize the appropriate treatment of disease. The aims of this paper are to review the recent world literature on chronic constipation and to propose a diagnostic and a therapeutic flow chart for this disorder. PMID- 11490800 TI - Using radionuclide esophageal emptying test to evaluate pneumatic dilatation effects for achalasia. AB - BACKGROUND/AIMS: Radionuclide esophageal emptying test using a solid meal is a simple, noninvasive, and physiologic test of esophageal function. METHODOLOGY: In this study, we evaluated esophageal emptying using an isotope-labeled solid meal (an egg salad sandwich labeled with 99mTc-MAA) in the assessment of pneumatic dilatation treatment in achalasia. Twenty achalasic patients (12 males, 8 females, age: 51.4 +/- 13.0 years) underwent esophageal emptying measurement before and after pneumatic dilatation. After treatment, the dysphagic symptoms of all patients improved. RESULTS: In comparison with the pretreatment test, the posttreatment retention fraction at 1, 5, 10, and 15 min were significantly lower (P < 0.001). CONCLUSIONS: The results suggest that radionuclide esophageal emptying test using a solid meal remains a useful objective study of esophageal function and may have an important future role in the follow-up evaluation of treatment for achalasia. PMID- 11490799 TI - Recurrent bleeding from a duodenal diverticulum 8 years after endoscopic treatment: case report and review of the literature. AB - A 70-year-old woman presented with a 2-day history of tarry stool. She had a history of hemorrhage from a duodenal diverticulum of the 2nd portion 8 years previously that had been managed successfully by endoscopic hemostasis. Initial gastrointestinal endoscopy revealed ulceration of the diverticulum with no active bleeding; nevertheless the ulceration was presumed to be the source of the tarry stool. Despite medical treatment, bleeding started again, but endoscopic ethanol injection achieved hemostasis. When bleeding started yet again 8 days after the endoscopic therapy, the patient underwent diverticulectomy. Although duodenal diverticula are frequently found in the adult gastrointestinal tract, they rarely show hemorrhage. Recently, there has been controversy about whether bleeding diverticula should be managed surgically or endoscopically. We describe for the first time a rare case of recurrent hemorrhage of a duodenal diverticulum after an 8-year interval; the case was treated by surgical diverticulectomy as a definitive therapy for the recurrent bleeding ulcer. We also present a review of the literature. PMID- 11490801 TI - Prevalence of esophagitis in H. pylori-positive peptic ulcer disease and the impact of eradication therapy. AB - BACKGROUND/AIMS: There have been recent reports of reflux esophagitis apparently occurring de novo after cure of H. pylori in peptic ulcer disease. The possibility that this phenomenon might be explained, at least in part, by unmasking of coexistent disease has not been assessed. The aim of this study was to assess the prevalence of esophagitis in H. pylori-positive peptic ulcer disease and examine the short-term impact of H. pylori therapy on the esophagus. METHODOLOGY: Esophagitis was systematically graded and the presence of hiatal hernia was noted in 244 peptic ulcer patients (duodenal 223; gastric 21) before and at least four weeks after triple therapy. H. pylori status was assessed using CLO test and histology, and esophagitis grade was assigned without knowledge of H. pylori status. RESULTS: Of the 244 patients, 49 (20%) had esophagitis which was grade 2 or more in over two-thirds. The prevalence of esophagitis was similar in duodenal and gastric ulcer patients. The presence of hiatal hernia was strongly associated with the finding of esophagitis (P < 0.001). Of 241 patients evaluable after therapy, 215 (89%) were H. pylori-negative and 26 remained H. pylori-positive. Esophagitis tended to improve or remain stable after H. pylori therapy and worsened in only 2 of the 49 patients (4%). Of 192 patients with a normal esophagus at baseline endoscopy, 14 (7%) showed evidence of esophagitis after therapy. The presence of hiatal hernia, but not cure of H. pylori, was significantly associated with the development of esophagitis. CONCLUSIONS: Our results indicate that esophagitis can coexist with peptic ulcer disease and persists after cure of H. pylori. Development of de novo esophagitis seems uncommon in the short-term after H. pylori therapy. Esophagitis in peptic ulcer disease is strongly associated with the presence of hiatal hernia. PMID- 11490802 TI - Obstruction due to persimmon bezoars: computed tomography detection. AB - BACKGROUND/AIMS: Small-bowel obstructions caused by persimmon bezoars are uncommon, and the diagnosis is difficult before operation. The aim of this study was to present computed tomography findings of persimmon bezoars in the small bowel. METHODOLOGY: We reviewed 3 cases diagnosed with small-bowel obstruction caused by persimmon bezoars between April 1997 and March 1999 at Chikamori Hospital. RESULTS: The cause of the small-bowel obstruction was bezoars in the 3 patients. Abdominal computed tomography performed before operation revealed a well-defined, intraluminal inhomogeneous mottled mass containing gas bubbles. CONCLUSIONS: Computed tomography is useful for diagnosing obstructed small-bowel persimmon bezoars. PMID- 11490803 TI - Surgery for Crohn's disease in Greece: a follow-up study of 79 cases. AB - BACKGROUND/AIMS: The aim of this prospective study was to elucidate the clinical features, indications for surgery and long-term outcome of a series of 79 Greek patients operated on for Crohn's disease. METHODOLOGY: The clinical features and follow-up of 79 out of 155 patients with definite diagnosis of Crohn's disease (51%), who were operated on at least once during the course of their disease, are analyzed. Three main locations of the disease were identified (small bowel, large bowel and small and large bowel). The need for surgery, indications for surgery and outcome of patients were analyzed and compared separately for these three main locations. The mean follow-up period after the first operation was 8.8 +/- 6.5 years. RESULTS: The proportion of men to women was 1.55:1 (P = 0.068). The main indication for surgery was poor response to conservative treatment, followed by obstructive ileus, erroneous diagnosis of acute appendicitis and development of fistulae or abscesses. Statistically significant differences between the three main locations of the disease were found for obstructive ileus (P < 0.01), and bowel perforation (P < 0.0297). Enterectomy and end-to-end anastomosis was the most frequently performed operation. Minor surgical procedures were performed mainly for drainage of perianal abscess. Differences in the number of operations required (one, two and three or more) according to the three main locations of the disease were statistically significant (P < 0.044). Emergency operation was required in 17.3%. Most of the urgently operated patients had only small bowel involvement. Twenty-six percent of patients required a surgical procedure for perianal disease. One or more, major or minor, perioperative complications occurred in 13 out of 79 operated patients (16.4%). No perioperative deaths were noticed. Evolution to cancer was observed in 2% (3 patients). The outcome of patients after the operation was characterized by exacerbations and remissions. A mortality rate of 11.6% was noticed in the follow-up period. However, most deaths were unrelated to the underlying Crohn's disease. CONCLUSIONS: The clinicoepidemiological characteristics of patients with Crohn's disease of Greek origin operated-on for their disease do not differ significantly from those reported from other Western or neighboring Mediterranean countries. However, other parameters such as the relatively low incidence of overall surgical need, the low incidence of colorectal cancer and the low incidence of surgery for perianal disease, all underline the importance of various genetic and environmental factors on the evolution and behavior of the disease in different parts of the world. PMID- 11490804 TI - A 3-day anti-Helicobacter pylori therapy is a good alternative for bleeding peptic ulcer patients with Helicobacter pylori infection. AB - BACKGROUND/AIMS: One-week triple therapy has been recommended as a standard regimen for eradicating Helicobacter pylori infection. The emergence of antibiotic-resistant strains, adverse drug effects, poor compliance and high cost of therapy add problems to the management of these patients. In this study, we assessed whether a 3-day triple therapy could be effective in eradicating Helicobacter pylori infection in bleeding peptic ulcer patients. METHODOLOGY: Peptic ulcer patients with Helicobacter pylori infection were enrolled in this study. Patients enrolled at the outpatient department (group A) received a 7-day oral regimen: bismuth subcitrate colloid 300 mg + amoxicillin 500 mg + metronidazole 250 mg four times per day. Patients who were admitted to the wards due to peptic ulcer bleeding (group B) received a 3-day regimen including omeprazole 40 mg intravenously every 6 hours, amoxicillin 500 mg + metronidazole 250 mg orally four times daily after hemostasis had been achieved. Patients of both groups received omeprazole 20 mg once per day or cimetidine 400 mg twice daily per os for at least-one month after anti-Helicobacter pylori therapy. We followed every patient endoscopically two months after anti-Helicobacter pylori therapy. RESULTS: From June 1997 to April 1999, a total of 57 patients (30 in group A and 27 in group B) with gastric or duodenal ulcer and Helicobacter pylori infection completed anti-Helicobacter pylori therapy. Two months after anti Helicobacter pylori therapy, peptic ulcer was found to be healed with a scar in 26 (86.7%) of group A and 23 (85.2%) of group B (P > 0.1). The eradication rates of Helicobacter pylori in the two groups were not significantly different in an intention-to-treat analysis [group A: 78.8% (26/33), 95% CI: 64.9-92.7%; group B: 80% (24/30), 95% CI: 65.7-94.3%, P > 0.1] and in a per protocol analysis [group A: 86.7% (26/30), 95% CI: 74.5-98.9%, group B: 88.9% (24/27), 95% CI: 77.1 100.7%, P > 0.1]. Fewer side effects occurred in group B (3/30) than those in group A (7/33) (P > 0.1). CONCLUSIONS: In patients with peptic ulcer bleeding a 3 day anti-Helicobacter pylori therapy is a good alternative for eradicating Helicobacter pylori infection. PMID- 11490805 TI - Role of upright chest radiography and ultrasonography in demonstrating free air of perforated peptic ulcers. AB - BACKGROUND/AIMS: The purpose of this study was to determine the value of upright chest radiography and ultrasonography in demonstrating free air of perforated peptic ulcers. METHOLOGY: Eighty-four patients with perforated peptic ulcers receiving both upright chest radiography and ultrasonography before laparotomy. The sensitivity of each modality in demonstrating free air was correlated. RESULTS: Among the 84 patients receiving both examinations, free air was demonstrated in only 39 (46%) upright chest radiographs and 46 (55%) ultrasonographs, the direct sign could be demonstrated in 57 (68%) patients by combined radiography and ultrasonography. Besides, the indirect sign of ascites could be demonstrated in 26 (31%) ultrasonographs. CONCLUSIONS: Ultrasonography is more sensitive than upright chest radiography to demonstrate free air of perforated peptic ulcers, and it should be considered in those patients of suspected perforated peptic ulcers with negative upright chest radiography. Combined methods of upright chest radiography and ultrasonography can increase the overall sensitivity in demonstrating free air. PMID- 11490806 TI - Regional chemoimmunotherapy for nonresectable metastatic liver disease of colorectal origin. A prospective randomized study. AB - BACKGROUND/AIMS: The study has been designed to assess the potential advantages of regional targeting chemoimmunotherapy versus systemic chemotherapy and immunotherapy in a number of patients suffering from nonresectable metastatic liver disease of colorectal origin. METHODOLOGY: From January 1996 to January 2001 (86) patients with unresectable metastatic liver disease of colorectal origin were randomly assigned to two groups. Group A (n = 44 patients), who received regional targeting chemoimmunotherapy through an arterial catheter introduced subcutaneously under echo guidance into the hepatic artery. Group B (n = 42 patients) received systemic chemoimmunotherapy. RESULTS: Survival of Group A patients ranged from 9 to 48 months (mean: 33 months), while Group B patients' survival ranged from 8 to 18 months (mean: 10 months). Response to treatment was 75% for Group A patients versus 52% for Group B. There were statistically significant differences in survival between the two groups (P = 0.0001). CONCLUSIONS: We believe that the data of this study reconfirms previously reported experience about the advantages of regional chemoimmunotherapy when compared with systemic chemoimmunotherapy for the management of advanced metastatic liver disease. PMID- 11490807 TI - Clinicopathological characteristics of patients with extrahepatic recurrence following a hepatectomy for hepatocellular carcinoma. AB - BACKGROUND/AIMS: None of the previous studies have compared the prognosis or clinicopathological factors between the patients with extrahepatic recurrence and those with intrahepatic recurrence of hepatocellular carcinoma after a hepatic resection. METHODOLOGY: The clinicopathological features and prognoses of patients with extrahepatic recurrence after a curative hepatectomy for hepatocellular carcinoma were investigated. RESULTS: Twenty-three patients with extrahepatic recurrence had more advanced-stage hepatocellular carcinoma at the primary operation compared to 186 patients with intrahepatic recurrence. After adjusting for tumor size, the prognosis of the 2 groups were comparable. However, among the patients with hepatocellular carcinoma exceeding 5 cm in diameter, the number of patients whose plasma levels of des-gamma-carboxy prothrombin was higher than 2.0 AU/mL in the patients with extrahepatic recurrence (62.5%) was significantly more (P < 0.05) than that in the patients with intrahepatic recurrence (20.0%). On the other hand, the prognosis of the 13 patients with extrahepatic recurrence alone was significantly better than in the 10 patients with both intrahepatic and extrahepatic recurrences. The prognoses of the 3 patients who underwent a surgical resection for isolated extrahepatic recurrence were markedly better than that of the remaining 10 patients only treated palliatively. CONCLUSIONS: If patients have tumors exceeding 5 cm in diameter and their plasma levels of des-gamma-carboxy prothrombin are higher than 2.0 AU/mL, more careful follow-up examinations than usual may thus be necessary in order to detect extrahepatic recurrence as early as possible. Furthermore, a surgical resection for the isolated extrahepatic recurrence of hepatocellular carcinoma is also recommended to produce long-term survivors. PMID- 11490808 TI - Management of intrabiliary ruptured hydatid disease of the liver. AB - BACKGROUND/AIMS: Intrabiliary rupture of hydatid cyst is an important complication of hydatid disease of the liver. The purpose of this study was to evaluate outcomes of patients with intrabiliary ruptured hydatid disease of the liver. METHODOLOGY: Two hundred and eleven patients were operated upon for hydatid cyst of the liver between 1990 and 1998 in our hospital. Twenty-four patients with intrabiliary rupture of hepatic hydatid cyst were retrospectively reviewed. RESULTS: Diagnosis of hydatid cyst was principally made using ultrasonography. We performed partial cystectomy + omentoplasty + transduodenal sphincteroplasty for 18 patients (75%). Daughter cysts and hydatid debris were found in common bile duct in 8 patients (33%). Internal opening of biliary fistula was found in 21 patients (87.5%). Morbidity rate was 12%. One patient died of unrelated causes. The average postoperative hospitalization period for the sphincteroplasty group was 7.6 days. CONCLUSIONS: Preoperative radiologic examination and laboratory values were not helpful to exclude an intrabiliary rupture of hydatid cysts in our patients. If bile stained cystic fluid is found, it is clear that a communication between hydatid cysts and the biliary tree exist. Transduodenal sphincteroplasty can be preferred in the management of intrabiliary ruptured hydatid disease because of short hospitalization, low morbidity, and ability to decompress intrabiliary pressure. PMID- 11490809 TI - Magnetic resonance cholangiopancreatography for diagnosing hepatolithiasis. AB - BACKGROUND/AIMS: Direct cholangiography with endoscopic retrograde cholangiopancreatography and percutaneous transhepatic cholangiography sometimes fails to adequately opacify the entire biliary tract, because of severe biliary obstruction caused by ductal stricture or lodged stones. We assessed the diagnostic accuracy of magnetic resonance cholangiopancreatography for hepatolithiasis. METHODOLOGY: Five patients with hepatolithiasis underwent ultrasonography, computed tomography, direct cholangiography, and magnetic resonance cholangiopancreatography, using a half-Fourier acquisition single-shot turbo spin-echo sequence. Surgical exploration or pathologic examination revealed stricture and dilatation of the intrahepatic ducts in all patients. Diagnostic accuracies for stones and ductal abnormalities were compared among the imaging studies. RESULTS: No complications occurred during magnetic resonance cholangiopancreatography studies. Magnetic resonance cholangiopancreatography fully depicted the biliary tract. Magnetic resonance cholangiopancreatography accurately detected and localized intrahepatic stones, as well as bile duct stricture and dilatation, in all patients. Intrahepatic stones were detected by endoscopic retrograde cholangiopancreatography in one of four patients and by percutaneous transhepatic cholangiography in all three who underwent this procedure. Endoscopic retrograde cholangiopancreatography and percutaneous transhepatic cholangiography demonstrated ductal stricture in all patients but failed to completely demonstrate the biliary tree in three of four patients, and one of three, respectively. On ultrasonography and computed tomography, precise localization of stones was difficult. Ultrasonography and computed tomography failed to demonstrate ductal stricture in one and two of the five patients, respectively. CONCLUSIONS: Magnetic resonance cholangiopancreatography diagnoses intrahepatic stones and bile duct abnormalities less invasively and more accurately than endoscopic retrograde cholangiopancreatography and percutaneous transhepatic cholangiography. PMID- 11490810 TI - Superoxide dismutase in hepatocellular carcinoma affects patient prognosis. AB - BACKGROUND/AIMS: The free radicals play an important role in the pathogenesis of neoplastic transformation of the tissues. Superoxide dismutase is a metalloenzyme, protecting the cells from oxygen radical insult. The superoxide dismutase activity may therefore alter the cellular signaling pathways against the insults derived from oxidative stress especially in the tumor tissues. Therefore, it is considered that superoxide dismutase activity is crucial in affecting the survival of the cancer bearing patients. This study aims to investigate the level of superoxide dismutase activity in hepatocellular carcinoma tissues and correlate this with patients' survival after surgery for hepatocellular carcinoma. METHODOLOGY: Thirty-six patients who had hepatectomy for hepatocellular carcinoma at the National Taiwan University Hospital from 1992 to 1993 were included in this study. Superoxide dismutase activity of the tumor tissues was determined. The results were correlated with the patients' survival. The patients were grouped based on their postoperative survival time. Those patients who were deceased less then one year after surgery were in group I. Group II included patients who survived more than one year but less than 3 years after operation. Group III patients survived more than 3 years but less than 5 years. Ten patients who survived longer than 5 years after surgical intervention were in group IV. Data were expressed as mean and analyzed with ANOVA. RESULTS: The demographic and clinical information of patients, such as age, gender, plasma albumin, globulin, alpha-fetoprotein levels, and hepatitis markers were comparable among these groups. The superoxide dismutase levels in the hepatocellular carcinoma were significantly higher in group IV than the other three groups (P < 0.05). Similarly, the superoxide dismutase levels of the hepatocellular carcinoma tissues from group III patients were significantly greater than those tissues from patients of either group I or group II. Tumor superoxide dismutase levels tended to be higher in group II than in group I, although it did not reach a statistical significance. CONCLUSIONS: Patients with higher superoxide dismutase levels in the hepatocellular carcinoma survived longer after hepatectomy. The superoxide dismutase levels of the tumor tissue may influence the malignancy and the outcome of the patients. It serves as prognostic factor for patients after hepatocellular carcinoma operation. PMID- 11490811 TI - Lactate levels in cirrhotic patients undergoing liver resection. AB - BACKGROUND/AIMS: The role of lactate in liver ischemia-reperfusion injury in cirrhosis has not been clarified. METHODOLOGY: One hundred patients with hepatocellular carcinoma who underwent partial liver resection under Pringle's maneuver were included in this study. Blood lactate was measured before the operation, three times during the surgery and on the first postoperative day to calculate its rate of accumulation or elimination. Aminotransferase levels were also recorded perioperatively. We calculated the rate of lactate accumulation during the pre-ischemic and ischemic phases and the elimination rate during the post-ischemic phase, and examined the correlation between these results and the clinical findings. RESULTS: The rate of lactate accumulation during the pre ischemic and ischemic phases was correlated with the preoperative indocyanine green retention rate (P = 0.04 and P = 0.004, respectively). The indocyanine green retention rate tended to be correlated with the rate of lactate elimination during the post-ischemic phase (P = 0.06). CONCLUSIONS: The blood lactate profile might be a reliable indicator of liver function during the perioperative period in cirrhotic patients who undergo liver resection with Pringle's maneuver. PMID- 11490812 TI - Treatment of hepatocellular carcinoma occurring after distal splenorenal shunt for esophagogastric varices. AB - BACKGROUND/AIMS: Whereas endoscopic therapy is hardly effective, distal splenorenal shunt is expected to have permanent hemostatic effects on the esophagogastric varices complicated with hepatocellular carcinoma and to sustain favorable general condition of the patient. In this study, we examined the effects of the shunt in the patients who developed hepatocellular carcinoma during the follow-up of the shunt operation. METHODOLOGY: Among the patients who had undergone distal splenorenal shunt operation for portal hypertension caused by cirrhosis, we selected only those who developed hepatocellular carcinoma during the follow-up, and then we reviewed our treatment of hepatocellular carcinoma. RESULTS: Hepatocellular carcinomas developed postoperatively in 12 out of 59 patients with the shunt operation. At onset of the carcinomas, the varices were well controlled with no risk of bleeding; and the liver function was reasonably maintained and pancytopenia was alleviated, compared to those at shunt operation. We performed hepatectomy in 4 cases and nonoperative therapies in 8 cases. After the therapies, no variceal bleeding occurred. Those therapies caused minor complications but no death. CONCLUSIONS: Distal splenorenal shunt is a useful therapy for postcirrhotic esophagogastric varices as it enables us to safely perform therapies for the hepatocellular carcinomas that develop during the follow-up period. PMID- 11490813 TI - Expression of matrix metalloproteinases in spontaneous regression of liver fibrosis. AB - BACKGROUND/AIMS: Spontaneous regression of liver fibrosis would depend on the degradation of the excessive matrix in the liver. In this study, we tried to determine the kinetics of the expression of genes for matrix metalloproteinase-2 and -13. METHODOLOGY: Liver fibrosis induced by carbon tetrachloride was resolved after withdrawal of this toxin. Histological staining for fibrous septa and determination of liver collagen content were used to evaluate the extent of liver fibrosis. Expression in liver of matrix metalloproteinase-2 and -13 was determined by reverse transcription-polymerase chain reaction. RESULTS: The fibrous septa became thinner and interrupted and liver fibrosis resolved rapidly within 10 days. Expression of matrix metalloproteinase-2 and -13 was elevated to 2.5- and 8.7-fold, respectively, at peak fibrosis. The former was maintained at 88%-76% and the later dropped rapidly to 30%-20% in the recovery periods. CONCLUSIONS: Resolution of liver fibrosis began within 10 days but only to 70%. Gene expression kinetics suggested metalloproteinase-13 might play a more important role in the resolution because it surged more markedly at peak fibrosis and returned to nearly basal levels in the recovery periods in parallel with liver collagen content. PMID- 11490814 TI - Inflammatory pseudotumors of the liver: prevalence and clinical impact in surgical patients. AB - BACKGROUND/AIMS: Progress in diagnostic imaging has increased the number of focal liver lesions detected and reports of an occasional finding of inflammatory pseudotumors of the liver are becoming numerous. To estimate their prevalence and clinical impact in surgical series we evaluate retrospectively our experience. METHODOLOGY: Four hundred and three patients carriers of a total of 717 focal liver lesions underwent liver resection consecutively in our Department from October 1995 to August 1999. All these patients underwent surgery. RESULTS: After surgical resection, 3 patients each proved to be carrying an IPT nodule accounting for 0.7% of all patients and 0.4% of all focal liver lesions. One inflammatory pseudotumor was only disclosed intraoperatively in a patient with an hepatocellular carcinoma. The other 2 accounted for 20% of the patients whose preoperative diagnoses were wrong. The operative procedures for the inflammatory pseudotumor nodules were: wedge resection, because the inflammatory pseudotumor was considered a new malignancy, a limited resection and a left extended hepatectomy with bilioenteric anastomosis, distal gastrectomy and lymphoadenectomy in one patient each. Inflammatory pseudotumors accounted for 33% of wrong indication for surgery. CONCLUSIONS: Our experience shows that, despite the low prevalence of hepatic inflammatory pseudotumors, their impact in the appropriate management of patients with focal liver lesions is not irrelevant. PMID- 11490815 TI - Long-term follow-up of interferon-treated chronic hepatitis C and serum hepatic fibrosis markers. AB - BACKGROUND/AIMS: We investigated the clinical application of serum fibrosis markers in a long-term follow-up of patients with chronic hepatitis C treated with interferon-alpha. METHODOLOGY: This study included 52 patients treated with interferon-alpha (total: 480 MU) for 6 months. They each underwent liver biopsy before and after treatment. Twenty-eight patients who underwent liver biopsy less than 2 years after treatment were classified as group 1, and 24 patients as group 2. The two groups were subdivided into HCV RNA-negative responders and HCV RNA positive nonresponders. Liver specimens were estimated using grading and staging scores. Serum hyaluronan, PIIIP, and type IV collagen levels were measured before and after treatment. RESULTS: In the responders of groups 1 and 2, grading score after treatment was significantly decreased compared with that before treatment. Staging score after treatment was significantly improved only in the responders of group 2. In the responders of group 2, serum hyaluronan level was significantly decreased compared with that before treatment. In group 2, the grading score was significantly correlated with serum PIIIP and type IV collagen levels, and the staging score was significantly correlated with only serum hyaluronan level. CONCLUSIONS: These findings indicate that the serum PIIIP and type IV collagen levels reflect the activity, and serum hyaluronan reflects the degree of fibrosis in liver specimens of HCV RNA-negative patients in a long-term follow-up of patients after interferon-alpha treatment. PMID- 11490816 TI - Hepatic resection for intrahepatic cholangiocarcinoma: relation to gross tumor morphology. AB - BACKGROUND/AIMS: The aim of this study was to investigate the clinicopathologic features and biological behaviors related to the gross appearance of intrahepatic cholangiocarcinoma. METHODOLOGY: Fourteen patients with intrahepatic cholangiocarcinoma who underwent hepatic resection between 1986 and 1998 were divided into four groups according to the gross appearance of the tumor: ID (intraductal growth) type (n = 1), PD (periductal-infiltrating) type (n = 4), MF (mass-forming) type (n = 5), MF-with-PD type (n = 4). RESULTS: Overall survival at 1, 5, and 10 years was 50.0%, 35.7%, and 35.7%, respectively. All three long term survivors without recurrence had tumors unassociated with vascular invasion, intrahepatic metastasis, or lymph node metastasis. The MF and MF-with-PD tumors were more frequently associated with vascular invasion and/or lymph node metastasis than the ID or PD type. The Ki-67-positive grade of the cancer cells was clearly higher in the MF and MF-with-PD tumors than in the ID or PD type. All of the cases of MF-with-PD tumors were stage IV-A and had a poor outcome. CONCLUSIONS: Extended hepatic resection with a sufficient surgical margin yielded good results in intrahepatic cholangiocarcinoma patients without vascular invasion, intrahepatic metastasis, or lymph node metastasis. However, it is necessary to develop a new effective strategy for advanced intrahepatic cholangiocarcinomas, such as the MF-with-PD type. PMID- 11490817 TI - Is mannitol effective against platelet-activating factor (PAF)-induced liver damage in obstructive jaundice? AB - BACKGROUND/AIMS: Platelet-activating factor, is a unique phospholipid with a broad range of biological activities that may be relevant in the development of inflammatory reactions. Platelet-activating factor has been suspected to play an important role in liver pathophysiology. The cultured Kupffer and endothelial cells produce and release platelet-activating factor in order to facilitate communication between hepatic sinusoidal and parenchymal cells. In this study, in the experimental jaundice model, platelet-activating factor levels were measured in liver tissue and plasma and the possible effects of mannitol on this mediator were assessed. METHODOLOGY: The experimental model consisted of 7 rats in the control group (CG), 7 rats in the sham operation group (ShG), and 7 rats in the obstructive jaundice group (JG) created by ligating the common bile duct. The last group was the mannitol-treated jaundiced group (MJG) and all animals in this group received 20% mannitol in doses of 2 mL/day, intraperitoneally, following common bile duct ligation. A week later all animals were sacrificed and plasma and liver tissue samples were collected. Platelet-activating factor levels were measured by radioimmunoassay technique. RESULTS: Liver tissue platelet activating factor levels (pg/mg tissue protein) were 72 +/- 18 in the CG, 183 +/- 51 in the JG, 84 +/- 17 in ShG, and 124 +/- 36 in MJG. Plasma levels were 460 +/- 13, 1600 +/- 40, 560 +/- 19, and 1200 +/- 23, respectively. In both sample types, MJG and JG values were significantly different from CG and ShG as well. MJG levels were also different from JG. CONCLUSIONS: These results showed that plasma and liver tissue platelet-activating factor levels are increased in experimental obstructive jaundice; and activation of this mediator contributes to the ongoing liver injury. Mannitol may improve or lessen this damage. PMID- 11490818 TI - Orthotopic liver transplantation in a young adult (33 years old) using an 84-year old donor. AB - Shortage of liver donors and the increasing number of patients on the waiting list for liver transplantation have led to a widening of the definition of liver donor suitability. Although the age limit for liver donors is controversial, current opinion is towards using liver allografts from donors older than 60 years. However, to date only a few cases that showed a good performance of liver graft by donors older than 60 years have been described. In this case report, orthotopic liver transplantation in a 33-year-old patient who received a graft from an 84-year-old donor is presented. A careful evaluation of the conventional donor-related risk factors (hemodynamics, hepatic function and histologic features) was carried out. Moreover, free radical scavenger glutathione was measured before cold ischemia and at the time of reperfusion in hepatic biopsies. After a 1-year follow-up, the recipient exhibits good general conditions and normal liver function values. PMID- 11490819 TI - Anomaly of the portal vein with an anomalous hepatic vein--the first case report. AB - An anomaly of the portal vein associated with an anomalous hepatic vein is described as the first reported case. A 44-year-old woman was incidentally found to have a huge hemangioma by ultrasonography. Computed tomography revealed an anomalous portal system with a normally located gallbladder and round ligament. Arterial portography revealed anomalous branching of the portal vein, with absence of the left umbilical portion, the curved right portal branch mimicked the right-sided umbilical portion. Hepatic venography demonstrated patent umbilical veins communicating with the left hepatic vein. Intraoperatively the hepatic vein branch was found on the surface of the right hepatic lobe and the common bile duct was dorsal to the hepatic artery. A limited excision of the involved liver was performed safely with the help of the preoperative definition of the abnormal liver anatomy. PMID- 11490820 TI - Vascular endothelial growth factors and liver diseases. AB - Vascular endothelial growth factor plays an important role in neovascularization both in normal tissues and most tumors. It has been extensively investigated recently in various hepatic diseases such as primary and secondary hepatocellular carcinoma, liver cirrhosis, hepatitis and even benign tumors in liver. Vascular endothelial growth factor has been verified to be closely involved in the development and metastases of hepatocellular carcinoma and correlated to the high risk of hepatic metastases and a poor prognosis in gastrointestinal cancer. Using antibodies to vascular endothelial growth factor or other drugs to suppress its expression has also been successfully tried to restrain hepatocellular carcinoma cells and metastases in vitro and in animal models. The protein of vascular endothelial growth factor has an inclination to increase in acute and chronic hepatitis and tends to decrease in cirrhosis both in tissue expression and circulating levels. This circulating level is closely related to the Child-Pugh classification in cirrhotic liver. However, there are indeed some disagreements concerning vascular endothelial growth factor and liver disease, for example, opinions on the positive rates of vascular endothelial growth factor in protein and mRNA level are far from reaching a general consensus. Further study should be performed in the future in antitumor research and its significance in the process of liver cirrhosis. PMID- 11490821 TI - Pancreaticoduodenectomy for periampullary cancer in patients more than 70 years of age. AB - BACKGROUND/AIMS: To assess the indications for and results of pancreaticoduodenectomy in patients more than 70 years old with periampullary cancer. METHODOLOGY: Thirty-four consecutive patients older than 70 years with periampullary cancer. The surgical procedure was pancreaticoduodectomy (Whipple's operation) with an extensive dissection of lymph nodes and the connective tissue in the peripancreatic region. Main outcome measures were postoperative morbidity and mortality, median and 5-year survival rates. RESULTS: Postoperative medical complications occurred in 24% and surgical complications in 53% of the patients. Four patients (12%) died in the postoperative period (within 30 days), and 3 patients (9%) died later in the postoperative course. The cumulative and age corrected 5-year survival rate for the remaining patients was 26%. Fifteen patients died of recurrence, and 7 patients of other causes. Five patients are still alive more than 5 years after surgery. In patients with noncurative operation the median survival time was 1 1/2 years, which is longer than would be expected from other palliative procedures. Apart from a moderately increased postoperative mortality the results were similar to those reported for younger patients. CONCLUSIONS: Pancreaticoduodenectomy should be considered in patients older than 70 years with resectable periampullary cancer. A 5-year survival rate of 20-35% can be obtained. Palliative resection may be indicated in patients in good general condition, as resection gives the best palliation and longer survival than other palliative methods. PMID- 11490822 TI - Long-term survival after surgical resection for pancreatic cancer. AB - BACKGROUND/AIMS: Pancreatic cancer remains one of the most formidable tumors defying early detection and effective treatment. Long-term survivors, however, do exist after resection. We investigated the clinicopathologic features of patients with pancreatic cancer who survived more than 5 years to draw out some suggestions concerning the indication of surgical treatment. METHODOLOGY: We studied the clinicopathologic features of 13 patients with pancreatic cancer who survived more than 5 years after resection. We reviewed their clinical records to investigate preoperative symptoms, serum tumor markers, operative findings, postoperative adjuvant therapy, and modes of recurrence and survival periods. Information on the location, size, histology and spread of the primary tumors were mainly obtained from pathology reports. RESULTS: Histologic types of the long survivors included ductal adenocarcinoma of common type in 4 patients, mucinous noncystic adenocarcinoma in 2, intraductal papillary-mucinous carcinoma (invasive) in 4, undifferentiated carcinoma in 1, endocrine tumor (islet cell carcinoma) in 1 and acinar cell carcinoma in 1. All 4 cases of ductal adenocarcinoma of the common type showed a moderate invasion either to the retroperitoneum, the portal vein or the duodenum. Two patients with mucinous noncystic carcinoma attained a long survival despite extensive invasion of the pancreatic stroma, although one died of peritoneal carcinomatosis. Two of 4 patients with intraductal papillary-mucinous cancer (invasive) died of peritoneal dissemination 6 and 11 years after resection, respectively. Three patients with cancer of other special histologic types, i.e., undifferentiated, well differentiated endocrine carcinoma and acinar cell carcinoma, showed invasion of the portal vein and splenic artery, involvement of the retroperitoneum and a metastatic tumor in the liver, respectively. CONCLUSIONS: Whereas special histologic types including ductal variants tended to predispose to long-term survival, ductal adenocarcinoma of the common type had some chance of long survival even with invasion of the surrounding tissues. PMID- 11490823 TI - Surgical treatment for mucin-producing tumors of the pancreas. AB - BACKGROUND/AIMS: Our objectives in this study were to evaluate the surgical treatment for mucin-producing tumor of the pancreas from the clinicopathological and imaging features. METHODOLOGY: Thirty-one patients with mucin-producing tumor of the pancreas were examined based on clinicopathological analyses to determine the appropriate surgical treatment. RESULTS: The clinical and imaging features easily distinguished the main duct type of intraductal papillary lesions (type Ia), branch type of intraductal papillary lesions (type Ib) and mucinous cystic neoplasms (type II). From pathological examinations, a dilated main pancreatic duct had the malignant potentiality and multicentric development. CONCLUSIONS: Pancreatic segments containing a dilated main pancreatic duct should be resected in type Ia. Type Ib is sufficient for partial resection without lymphadenectomy. Type II also requires partial resection of the cystic neoplasm. A standard lymphadenectomy may be an option when type Ia and II show invasive features. PMID- 11490824 TI - Correlation of postoperative survival and angiogenic growth factors in pancreatic carcinoma. AB - BACKGROUND/AIMS: VEGF (vascular endothelial growth factor) and EGF (epidermal growth factor) are promoters of angiogenesis. It was the aim of this study to investigate a possible coexpression of both growth factors in tumor samples of pancreatic cancer patients in relation to survival after resection of the tumor. METHODOLOGY: We investigated the expression of VEGF165 and EGF in tumor specimen from 19 patients that underwent pancreaticoduodenectomy. Growth factor expression was determined using immunohistochemical methods. RESULTS: Coexpression of VEGF165 and EGF was observed in tumor samples of 9 (47%) patients. VEGF165 and EGF expression in the same tumor correlates significantly (P < 0.05, Fisher test). UICC stage III pancreatic carcinoma patients with VEGF165 negative tumor cells had a significantly better outcome after surgery compared to UICC stage III patients with VEGF165-positive tumor cells (median survival time 19 months vs. 9 months respectively; P < 0.05, Wilcoxon-test). CONCLUSIONS: Antiangiogenic therapy after surgery for pancreatic cancer may be beneficial, especially for UICC III patients. PMID- 11490826 TI - Hypersplenism induced by hepatectomy. AB - BACKGROUND/AIMS: We encountered a case of posthepatectomy splenic enlargement and hypersplenism followed by disseminated intravascular coagulopathy with airway hemorrhage causing death. METHODOLOGY: We, therefore, retrospectively investigated postoperative splenic enlargement, hypersplenism and disseminated intravascular coagulopathy by computed tomography and laboratory data in 57 hepatectomized patients with a malignant or benign disease in the postoperative period. RESULTS: Of 32 patients with hepatocellular carcinoma or biliary tract carcinoma (group A), 12 with metastatic hepatic lesions (group B), and 13 with benign liver disease (group C); remarkable (20%) splenic enlargement was noted in 8 patients in group A, 2 in group B, and 2 in group C. Seven of the 12 patients were associated with liver cirrhosis, 5 with preoperative splenomegaly, and 8 had undergone major hepatectomy. Postoperative hypersplenism developed in 5 patients in group A, and one patient in group C. All of them were associated with liver cirrhosis or chronic hepatitis and preoperative splenomegaly, and five had undergone hepatic lobectomy or more extensive resections. All except for the disseminated intravascular coagulopathy case recovered. Statistically, splenic enlargement was significantly related to the extent of hepatectomy; lobectomy versus segmentectomy = 28.3 +/- 28.5% (n = 14) versus 12.4 +/- 13.8% (n = 20), (unpaired Student's t test, P = 0.037). Platelet counts of the patients with liver cirrhosis or chronic hepatitis is lower than those without the diseases, both pre- and postoperatively (14.0 +/- 6.0 x 10(4)/mm3 vs. 21.5 +/- 6.2 x 10(4)/mm3, P = 0.0001). CONCLUSIONS: Postoperative hypersplenism was noted only in the patients with liver cirrhosis or chronic hepatitis and preoperative splenomegaly, and developed more frequently after larger hepatectomies than after smaller hepatectomies; 5 (45%) of 11 versus 1 (7%) of 14, chi 2 test, P = 0.026). PMID- 11490825 TI - Right hepatic artery interruption and prostaglandin E1 in total or proximal pancreatectomy for pancreatobiliary malignancy. AB - BACKGROUND/AIMS: Advanced hepato-biliary-pancreatic malignancy can frequently involve the hepatic artery. We evaluated the use of prostaglandin E1 in total or proximal pancreatectomy with the right hepatic artery interruption. METHODOLOGY: A Consecutive seven of 117 patients (6.0%) in whom the right hepatic artery was interrupted and not reconstructed were reviewed retrospectively. Four of them received prostaglandin E1 (10-20 ng/kg/min) until the fifth postoperative day, while, the remaining three did not. The effect of prostaglandin E1 was compared concerning complication and hepatic function. RESULTS: The right hepatic artery was intentionally resected because of cancer invasion in five patients with biliary tract carcinoma, while, accidentally transected in two with pancreatic carcinoma. Operative deaths did not occur. The biliary leakage was identified in one patient treated without prostaglandin E1. Although a marked rise in glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and lactate dehydrogenase levels was observed, hepatic dysfunction was successfully treated conservatively in all patients. The glutamic oxaloacetic transaminase and lactate dehydrogenase values were significantly lower (P < 0.05) in patients treated with prostaglandin E1 compared with those without prostaglandin E1. CONCLUSIONS: The prostaglandin E1 infusion can be helpful for biliary anastomosis and hepatic function in radical hepato-biliary-pancreatic surgery with the right hepatic artery interruption. PMID- 11490827 TI - Treatment of Helicobacter pylori in patients with lymphocytic gastritis. AB - BACKGROUND/AIMS: Lymphocytic gastritis is a subtype of chronic gastritis characterized by a marked increase in the number of intraepithelial lymphocytes in the gastric mucosa. Its etiology is unknown, but a proportion of these patients have Helicobacter pylori infection. The aim was to assess the significance of H. pylori treatment in lymphocytic gastritis patients. METHODOLOGY: The 10 patients with lymphocytic gastritis and either serologically or histologically diagnosed H. pylori infection were treated with a triple therapy and followed by serology and histology after 6-18 months. RESULTS: The levels of IgG antibodies for H. pylori decreased below 50% of the pretreatment values in all patients. The maximum numbers of intraepithelial lymphocytes decreased significantly (P = 0.005) from the pretreatment values. CONCLUSIONS: Treatment of H. pylori infection cures lymphocytic gastritis associated with H. pylori infection. H. pylori appears to be one etiological cause of lymphocytic gastritis. PMID- 11490828 TI - Improvement of the prognosis of gastric cancer with extensive serosal invasion using left upper abdominal evisceration. AB - BACKGROUND/AIMS: The prognosis of gastric cancer patients with serosal invasion is very poor. In this study, the effectiveness of the LUAE (left upper abdominal evisceration) procedure for these patients was evaluated retrospectively. METHODOLOGY: Thirty-seven gastric cancer patients who had serosal invasion but no massive peritoneal metastasis or hepatic metastasis, and underwent LUAE, were enrolled in this study (LUAE group). As a control, 66 gastric cancer patients who had the same disease conditions as the LUAE group, and underwent conventional total gastrectomy with the combined resection of the pancreatic body and tail and spleen (TPS group), were also investigated. RESULTS: The survival rate (5-year, 42.2%) of the LUAE group was significantly better than that (5-year, 21.2%) of the TPS group (P = 0.009). Although D4 super-extended lymphadenectomy and intraperitoneal chemotherapy during surgery was performed more frequently in the LUAE group than those in the TPS group, multivariate analysis demonstrated that the LUAE procedure was a better independent prognostic factor. CONCLUSIONS: The LUAE procedure in combination with D4 super-extended lymphadenectomy and intraperitoneal chemotherapy improved the prognosis of gastric cancer patients with extensive serosal invasion. PMID- 11490829 TI - Delayed gastric emptying in subjects with Gilbert's syndrome. AB - BACKGROUND/AIMS: Recently, it has been proposed that decreased intestinal motility in fasting-induced hyperbilirubinemic rats probably results in an increase in the enterohepatic cycling of unconjugated bilirubin. We investigated the association among gastric emptying, intestinal transit time, and serum unconjugated levels in subjects with Gilbert's syndrome. METHODOLOGY: Ten subjects with Gilbert's syndrome were included in this study according to the following criteria: fasting hyperbilirubinemia; no hemolysis or gastrointestinal disorders and free of any medication. Five normal, healthy volunteers acted as controls. Gastric emptying and intestinal transit time were evaluated after overnight fasting by administration of a standard meal mixed with 1-2ci of 99Tc labeled diethylene-triamine-pentacetic acid. Serum unconjugated bilirubin levels were determined by high-performance liquid chromatography. RESULTS: The gastric emptying in Gilbert's syndrome subjects was 134.1 +/- 38.9 and 90.9 +/- 6.5 min in controls, P < 0.03. It was a tendency to have a shorter intestinal transit time in subjects with Gilbert's syndrome, 138.3 +/- 59.0, than in control subjects, 183.8 +/- 11.3 min. Serum unconjugated bilirubin levels (mg/dL) were 2.6 +/- 1.04 and 0.95 +/- 0.34, P < 0.01. CONCLUSIONS: Gastric emptying is delayed significantly in Gilbert's syndrome, and intestinal transit time differences between Gilbert's syndrome subjects and controls were not significantly different. PMID- 11490830 TI - A novel operative technique on proximal gastrectomy reconstructed by interposition of a jejunal J pouch with preservation of the vagal nerve and lower esophageal sphincter. AB - BACKGROUND/AIMS: This article describes the surgical techniques and postoperative status for proximal gastrectomy reconstructed by interposition of a jejunal J pouch with preservation of the vagal nerve and lower esophageal sphincter. METHODOLOGY: We have performed a new technique for reducing postgastrectomy sequelae such as reflux esophagitis, early dumping syndrome, and microgastria in early gastric cancer located in the proximal third of the stomach. The technique consists of proximal gastrectomy with preservation of the hepatic, pyloric, celiac branch of the vagal nerve, and abdominal esophagus (lower esophageal sphincter), and reconstruction by interposition of a jejunal J pouch. To reserve pyloric function, pyloroplasty can be omitted by preservation of the pyloric branch from the vagal nerve. To restore loss of reservoir function, the reconstruction is performed with an interposed jejunal J pouch. Sacrifice of the mesenteric arcades is kept to a minimum to preserve the autonomic nerve and blood flow in the mesentery. RESULTS: All of the patients who underwent this operation were able to eat an adequate amount of food at 6 months after surgery and they were satisfied with their postoperative status. And that, we have not experienced postgastrectomy disorders such the dumping syndrome and reflux esophagitis. CONCLUSIONS: Therefore, this method is useful for preventing the postoperative disorders in patients with early gastric cancer located in the proximal third of the stomach. PMID- 11490831 TI - Is the local acid output an important factor for Helicobacter pylori colonization? AB - BACKGROUND/AIMS: Low local acid production is considered to be an important factor for Helicobacter pylori colonization. The increased bacterial density and inflammation are expected in cardiac mucosa because of low acid secretion. We aimed to investigate the bacterial density and the histologic pattern of gastritis in the cardia, and to compare with those of the antrum and corpus. METHODOLOGY: The biopsy specimens taken from the antrum, corpus and cardia in 97 patients during endoscopy were examined histopathologically. RESULTS: The mean scores for bacterial density in the cardia, corpus and antrum were 1.32 +/- 0.35, 1.09 +/- 0.86, 1.42 +/- 0.97, respectively. Bacterial density of the cardia was significantly higher than that in corpus, but was similar to the antrum. The mean scores for the degree of gastritis in the cardia, corpus and antrum were 1.56 +/- 0.79, 1.64 +/- 0.75, 1.85 +/- 0.77, respectively. Gastritis was significantly more severe in the antrum than in the corpus and cardia. The activity scores of gastritis in the three regions were statistically similar. CONCLUSIONS: Although bacterial density in the cardiac mucosa were found to be high, the degree of gastritis was low. PMID- 11490832 TI - Endoscopic treatment of hemorrhagic gastric ulcer in patients aged 80 years or more. AB - BACKGROUND/AIMS: Hemorrhagic gastric ulcer is a more serious disorder in elderly patients than in young patients. This study aimed to evaluate the efficacy of endoscopic treatment for hemorrhagic gastric ulcer in patients aged 80 years or more. METHODOLOGY: A total of 269 patients underwent endoscopic treatment for hemorrhagic gastric ulcer with hemoclipping and pure ethanol injection, either alone or in combination. These patients were divided into two groups: elderly (> or = 80 years old) and younger (< 80 years) groups. The clinical data, endoscopic findings and outcomes of endoscopic treatment were compared between these two groups. RESULTS: Patients in the elderly group had a significantly higher incidence of concomitant disease (73% vs. 23%), anemia (7.9 +/- 2.1 g/dL vs. 9.3 +/- 2.8 g/dL) and large ulcer (23% vs. 6%) compared with the younger group. However, all patients in the elderly group underwent endoscopic hemostasis successfully with no complications and no deaths. The rebleeding rate was not significantly different (8% vs. 4%) between the two groups. CONCLUSIONS: Elderly patients with hemorrhagic gastric ulcer have high incidences of severe ulcer disease and concomitant medical problems. Endoscopic hemostasis for hemorrhagic gastric ulcer is effective and safe, even for such elderly patients. PMID- 11490833 TI - Serum levels of interleukin-2 and tumor necrosis factor-alpha correlate to tumor progression in gastric cancer. AB - BACKGROUND/AIMS: The aim of this study was to assess serum levels of Interleukin 2 and tumor necrosis factor-alpha with disease progression and correlate these levels with CEA and CA19-9 serum levels. METHODOLOGY: Serum levels of interleukin 2 and tumor necrosis factor-alpha were measured in 23 patients with gastric cancer (being 9 stage I or II and 14 stage III or IV) and 10 patients without cancer by ELISA using Predicta Genzyme Diagnostica. The patients were followed for at least 2 years or until death. CEA and CA19-9 were also measured in both groups by ELISA (Abbott Diagnostic). RESULTS: Patients with gastric cancer stage III or IV had elevated levels of these cytokines (P = 0.002 for IL-2 and P = 0.003 for tumor necrosis factor-alpha). There was no difference between the serum levels of tumor necrosis factor-alpha and interleukin-2 in patients with gastric cancer stage I or II and the control group (P > 0.05). We also found no difference among the groups for CEA and CA19-9 (P = 0.17 and 0.72, respectively). Only one gastric cancer patient stage I or II had elevated level of IL-2 and none had elevated levels of tumor necrosis factor-alpha. In the group of patients with gastric cancer stage III or IV, 87.5% of them with elevated levels of tumor necrosis factor-alpha and 75% of them with elevated levels of interleukin-2 died during the follow-up. CONCLUSIONS: We conclude that serum interleukin-2 and tumor necrosis factor-alpha are associated with advanced gastric cancer and that these cytokines might be a useful tumor marker for gastric cancer, being associated with poor prognosis. PMID- 11490834 TI - Survival after curative resection for stage IE and IIE primary gastric lymphoma. AB - BACKGROUND/AIMS: There is considerable controversy regarding the optimal treatment of patients with primary gastric lymphomas. However, surgery still plays an important role in the management of stage IE and IIE gastric lymphomas. We aimed at assessing survival of primary gastric lymphoma cases with stage IE or IIE that were surgically treated at the Surgical Oncology Department. METHODOLOGY: Thirty-seven patients with stage IE and IIE primary gastric lymphoma who were surgically treated and had complete follow-up from January 1990 to September 1998 were reviewed retrospectively. Patients' age, gender, tumor location, tumor grade, histologic type, depth of tumor invasion, regional lymph node status, tumor stage, type of gastrectomy (total/subtotal), combined resection, extensive lymphadenectomy, adjuvant chemotherapy were used as the clinicopathologic variables. RESULTS: Five-year survival rates for stage IE and stage IIE disease were 75% and 37%, respectively. The overall 5-year survival rate of the patients was 57%. Univariate analysis demonstrated that age, tumor stage, and type of gastrectomy were associated with prognosis, but only type of gastrectomy (subtotal gastrectomy) and tumor stage were found to be independent prognostic factors (P < 0.05). CONCLUSIONS: To obtain prolonged survival we recommend radical resection with extensive lymphadenectomy for malignant lymphoma stages IE and IIE. Patients with small distal lymphomas of the stomach can be treated with subtotal gastric resection. PMID- 11490835 TI - A case of AFP-producing early gastric carcinoma with rapid growth liver metastasis. AB - An 84-year-old man presented with complaints of epigastric discomfort. Upper gastrointestinal series and endoscopy showed an elevated lesion at the posterior wall of greater curvature on the gastric fundus. Diagnosed as moderately differentiated tubular adenocarcinoma by biopsy, wedge resection and 4sa regional lymph node dissection were carried out. The tumor morphology showed type I with slight elevation, 2.5 x 1.7 cm in size; histological showed papillary, tubular, and solid formations having clear cytoplasm and large bizarre nuclei invading the deep submucosal layer (sm2). This case was evaluated as T1(sm) N0 M0 stage Ia early gastric cancer. In the 5th month after operation, multiple liver metastases were detected. He died of liver failure by rapid growth of metastatic tumors in the 6th month after operation. The serum alpha-fetoprotein level at recurrence was 1,900 ng/mL, and alpha-fetoprotein-positive cells were immunohistochemically detected in operative and liver biopsy specimens. PMID- 11490836 TI - Gastric sarcoidosis--a single polypoid appearance in the involvement. AB - We describe a case of gastric sarcoidosis which developed during a 10-year period of observation of generalized sarcoidosis. The patient was asymptomatic, but gastroduodenoscopy revealed a polypoid lesion in the antral region. The specimen obtained by polypectomy showed noncaseating granuloma, suggesting sarcoidosis. Chest radiograph and computed tomographic examination revealed no involvement of the lung or mediastinal lymph nodes, although these findings were present at the initial diagnosis. The present case indicates that gastric involvement should be considered in patients with sarcoidosis, even when no hilar lymphadenopathy is present. Furthermore, the macroscopic appearance of a single polyp in gastric sarcoidosis is extremely rare in gastric sarcoidosis in the literature. PMID- 11490837 TI - Krukenberg tumor from gastric mucosal carcinoma without lymphatic or venous invasion: report of a case. AB - A 49-year-old woman was admitted because of early gastric carcinoma. Subtotal gastrectomy was performed. In the resected specimen, gastric mucosal carcinoma without lymph node metastases was located in the prepyloric region. Histologic type was moderately differentiated adenocarcinoma and signetring cell carcinoma, and there was no lymphatic or venous invasion. One year after operation, a left ovarian tumor was detected. At the second operation, bilateral oophorectomy and hysterectomy were performed. Pathological findings revealed Krukenberg tumors originating from the gastric carcinoma in the bilateral ovaries. One year after the second operation, a hard mass due to cancer recurrence developed in the pelvis with symptoms including tenesmus and abdominal pain. Chemotherapy and palliative colostomy were performed. She died of peritonitis carcinomatosa six years and two months after the first operation. We experienced a rare case of Krukenberg tumor with two interesting points; its origin was gastric mucosal carcinoma without lymphatic or venous invasion, and the patient survived for more than four years after the diagnosis. PMID- 11490839 TI - Recent advances in diagnosis and treatment of pancreatic cancer. PMID- 11490838 TI - Extreme hypergastrinemia caused by atrophic gastritis and Helicobacter pylori infection--a case report. AB - We present a case with extremely high serum gastrin induced by atrophic gastritis and Helicobacter pylori infection. The patient, a 95-year-old male, was diagnosed with idiopathic chronic diarrhea. During diagnostic work-up, his fasting serum gastrin was up to 2078 pg/mL. The secretin test was negative for gastrinoma. Octreotide scan showed no suspicious lesion except for diffuse faint uptake over the gastric antrum on the 48-hour-delay film. Gastric acidity test revealed achlorhydria. On histology examination, atrophic gastritis with Helicobacter pylori infection was found in the gastric body, but the antral mucus was normal with a slight increase in gastrin-secreting cells. To our knowledge, such extremely high serum gastrin induced by atrophic gastritis and Helicobacter pylori infection has never been reported before. PMID- 11490840 TI - Recent advances in US diagnosis of pancreatic cancer. AB - Recent years have seen dramatic developments in extracorporeal ultrasonography: Color, including-power Doppler ultrasonography, ultrasonography angiography, harmonic imaging (tissue harmonic imaging and contrast harmonic imaging) and 3 dimensional ultrasonography (including virtual endoscopy). In this report, we describe the present situation and the prospective outlook for these new diagnostic modalities mainly on the basis of our own experiences. PMID- 11490841 TI - CT and MR imaging of pancreatic cancer. AB - Recent improvements in imaging techniques have made it possible to improve the diagnostic accuracy for detection, staging, and indicating surgical resectability of pancreatic cancer. The latest advance in the computed tomography technique, is the introduction of subsecond multislice helical scanning that improves z-axis resolution in the reformatted images and three-dimensional rendering with a large volume data. Magnetic resonance imaging provides versatile information including magnetic resonance cholangiopancreatography that allows noninvasive delineation of the pancreatic and biliary duct systems. The presence of pancreatic cancer may best be evaluated by dynamic computed tomography or dynamic magnetic resonance imaging with administration of intravenous contrast material. Both computed tomography and magnetic resonance imaging are valuable for the preoperative assessment of local invasion and vascular involvement. Multislice helical computed tomography is currently considered as the best single modality for the diagnosis of pancreatic cancer as it provides excellent image quality. When advanced magnetic resonance imaging equipment is used as a primary modality, in the future, it may have a possibility to replace other imaging modalities. PMID- 11490842 TI - Endoscopic diagnosis of pancreatic cancer using intraductal ultrasonography. AB - BACKGROUND/AIMS: At present developed modalities are not sufficient for detecting early stage pancreatic cancer. We previously reported the clinical usefulness of intraductal ultrasonography in various pancreatobiliary diseases. In the present study we assessed the usefulness of intraductal ultrasonography in diagnosing pancreatic cancer. METHODOLOGY: Thirty-one patients with pancreatic cancer were examined by intraductal ultrasonography. We approached the main pancreatic duct (pancreatic duct-intraductal ultrasonography) in 24 of 31 patients and the bile duct (bile duct-intraductal ultrasonography) in 20 patients with pancreatic cancer. We compared the diagnostic ability of pancreatic duct-intraductal ultrasonography with that of extracorporeal ultrasonography, computed tomography, endoscopic ultrasonography or endoscopic retrograde pancreatography. We examined the usefulness of bile duct-intraductal ultrasonography in diagnosing tumor invasion to the bile duct. RESULTS: Pancreatic duct-intraductal ultrasonography was able to demonstrate a tumor in 22 of 24 patients. Extracorporeal ultrasonography, computed tomography, endoscopic ultrasonography or endoscopic retrograde pancreatography detected tumors in 26, 27, 29, 29 of 31 patients, respectively. In two patients, only intraductal ultrasonography could demonstrate a tumor, which was not detected by any other modalities. We examined bile duct invasion of the tumor according to our grading system. The overall accuracy rate was 90%. No complications were noted in any patients throughout the study period. CONCLUSIONS: Intraductal ultrasonography is useful to diagnose pancreatic cancer, and it is suggested that it should be actively performed after endoscopic retrograde pancreatography. PMID- 11490843 TI - Molecular diagnosis of pancreatic cancer. AB - Pancreatic ductal adenocarcinoma is a result of accumulated genetic alterations, including oncogenes such as K-ras, tumor-suppressor genes such as p53, p16 and DPC4 and genome-maintenance genes such as BRCA2, microsatellite instability and telomerase. Recent findings which characterize the molecular genetic profile of the pancreatic cancer have reshaped the nomenclature describing histological progression in pancreatic ductal tumorigenesis. K-ras mutations frequently occur early, whereas changes in the expression and genetic integrity of the p16 gene appear in intermediate lesions, and the inactivation of the p53, DPC4 genes and activation of telomerase occur late in the neoplastic progression. So far K-ras and telomerase activity have been used as molecular markers for the diagnosis of pancreatic carcinoma, whereas p53 and p16 may be a prognostic indicator of pancreatic cancer. Additional tumor-suppressor genes and the related signaling pathway such as ALK-5 are likely to be defined. In addition to the human genome project, these new advances hopefully will accelerate the development of diagnostic and screening techniques for this grave condition. PMID- 11490844 TI - Clinical usefulness of the immunostaining of the tumor markers in pancreatic cancer. AB - The effect of the rapid immunostaining of gastrointestinal cancer-associated antigens, CA19-9, CEA, DUPAN2, and CA50 was discussed for intraoperative pathological diagnosis of pancreatic cancer. The method can be completed in only 13 minutes with microwave irradiation to accelerate the incubation of the primary antibody. Only 3 seconds of irradiation at 500 W for fresh-frozen sections produced specific antigen staining of greater intensity than that obtained with longer incubation by the conventional method. Preservation of the tissue structure was satisfactory with minimal nonspecific background staining enabling us to diagnose the intrapancreatic spread of cancer. This method was also applied to intraoperative peritoneal washing cytology. As with frozen section biopsy, the sensitivity of intraoperative cytology is greater than by the conventional staining method, which is able to achieve more precise staging of pancreatic cancers. Our rapid immunoperoxidase staining method on the cryostat section of pancreatic biopsy specimens and on cytology samples provides important information to determine an appropriate operative approach for pancreatic cancer. PMID- 11490845 TI - Intraoperative diagnosis of pancreatic cancer extension using IVUS. AB - BACKGROUND/AIMS: Pancreatic cancer easily invades retroperitoneal tissue, especially the portal vein and extrapancreatic nerve plexus. We evaluated the diagnostic accuracy of intraportal endovascular ultrasonography in portal vein and extrapancreatic nerve plexus invasion. METHODOLOGY: Intraportal endovascular ultrasonography was performed in 78 cases of pancreatic cancer (head 67, body 8, total 3). Intraportal endovascular ultrasonography was performed intraoperatively from the superior mesenteric vein with an 8-French, 20-MHz intravascular ultrasound catheter. Three-dimensional intraportal endovascular ultrasonography was constructed by volume rendering. RESULTS: Intraportal endovascular ultrasonography visualized the portal vein as an echogenic band with a thickness of 0.5 mm to 1.0 mm. The diagnostic criterion of portal vein invasion was obliteration of this echogenic band. Intraportal endovascular ultrasonography visualized segment II of the extrapancreatic nerve plexus as the high-echoic area around the inferior pancreaticoduodenal artery. The diagnostic criterion of extrapancreatic nerve plexus invasion was low-echoic infiltration around the inferior pancreaticoduodenal artery. The sensitivity, specificity, and overall accuracy of intraportal endovascular ultrasonography for diagnosis of portal vein invasion was, respectively, 97.4%, 92.5%, and 94.9%. The values for diagnosis of extrapancreatic nerve plexus invasion, respectively, were 94.4%, 97.1%, and 96.2%. Three-dimensional intraportal endovascular ultrasonography depicted the invasion area as a defect of the portal vein wall. CONCLUSIONS: Intraportal endovascular ultrasonography detected subtle portal invasion and provided accurate portal invasion area which was useful for portal vein an reconstruction. Intraportal endovascular ultrasonography could also diagnose the extrapancreatic nerve plexus invasion of segment II. PMID- 11490846 TI - The role of extended radical operation for pancreatic cancer. AB - BACKGROUND/AIMS: To clarify the indication of extended operation for pancreatic carcinoma, a clinical study was carried out. METHODOLOGY: From July 1981 to 1999, 196 of 307 (63.8%) patients with pancreatic carcinoma underwent resection of the tumor. Portal vein resection was performed in 145 of these 196 (74.0%) resected cases. The postoperative survival rate was studied according to the operative and histopathological findings. RESULTS: In spite of the aggressive surgery, there was no patient who survived over 3 years after operation in the group carcinoma positive on the surgical margins. Patients who survived over 3 years postoperatively were observed in the group of carcinoma-free surgical margins. CONCLUSIONS: The most important indication of extended operation combined with portal vein resection for pancreatic cancer is to obtain surgical cancer-free margins. There is no indication of extended operation for cases in which surgical margins will become cancer-positive, if such an operation is employed. PMID- 11490847 TI - The role of adjuvant therapy for pancreatic cancer. AB - BACKGROUND/AIMS: In spite of radical pancreatectomy with lymphadenectomy for adenocarcinoma of the pancreas, survival remains poor. We add two forms of adjuvant therapy, i.e., intraoperative radiotherapy and liver perfusion chemotherapy, to the radical resection. The aim of this study is to investigate the utility of these two forms of adjuvant therapy prospectively and to review their effectiveness. METHODOLOGY: One hundred and ninety-six patients with pancreatic cancer who underwent radical pancreatectomy in our institute were enrolled. We conducted intraoperative radiation therapy 30 Gy against the retroperitoneal connective tissues around the superior mesenteric artery during surgery. Adjuvant liver perfusion chemotherapy was added immediately after operation via the portal vein using 5-fluorouracil (250 mg/body/day) for 3 to 4 weeks continuously. Overall survival analyses were done by the method of Kaplan and Meier. RESULTS: Intraoperative radiation therapy did not influence the prognoses directly. However, the patients who received adjuvant liver perfusion chemotherapy had better prognoses (P < 0.05). CONCLUSIONS: Although adjuvant therapy after radical resection with wide lymphadenectomy improves prognoses, the results are still not satisfactory. We should develop a new and more efficacious treatment. PMID- 11490848 TI - Gene therapy for pancreatic cancer. AB - Despite improvements in surgical care and locoregional therapy, the prognosis of patients with pancreatic cancer has seen little improvement over the last several decades. It is difficult to diagnose pancreatic cancer at its earliest stages when it is amenable to cure by surgical resection because it is too small to produce symptoms in the affected patient. Recent improvements in radiographic modalities aimed at earlier detection and extent of cancer spread have enabled the clinician to provide the most efficacious treatment regimen possible. Nevertheless, pancreatic cancer is very aggressive locally and frequently metastasizes to the liver and peritoneum. New strategies are necessary to treat pancreatic cancer and gene therapy offers hope in this regard. Many studies have revealed the promise of gene therapy in the treatment of pancreatic cancer in rodent models. Early clinical trials are ongoing to evaluate the success of these gene therapy regimens in humans. In this article we review the gene therapy strategies currently employed in the fight against pancreatic cancer, including antisense strategies, gene-directed prodrug activation therapy, promoter gene strategies, and oncolytic viral therapy. PMID- 11490849 TI - Evaluation of various imaging methods in the differential diagnosis of intraductal papillary-mucinous tumor (IPMT) of the pancreas. AB - BACKGROUND/AIMS: IPMT (intraductal papillary-mucinous tumor) of the pancreas has unique clinicopathological characteristics. The lesions which show characteristic clinical features of IPMT exhibit a wide spectrum of histological types ranging from atypical hyperplasia to invasive cancer. Therefore, surgical treatment cannot be recommended for all patients with IPMT. It is necessary to assess the malignant potential of IPMT in individual patients in order to select an appropriate approach. The aim of this study was to evaluate the effectiveness of endoscopic ultrasonography and intraductal ultrasonography as compared with ultrasonography and computed tomography for this purpose. METHODOLOGY: Ultrasonography, computed tomography, endoscopic ultrasonography and intraductal ultrasonography were performed in 49 cases of IPMT (atypical hyperplasia 7, adenoma 23, noninvasive 7 and invasive adenocarcinoma 12). On the basis of the histopathological analysis of another 28 cases of resected IPMT specimens, criteria for differential diagnosis by imaging modalities were defined as follows: Nonneoplastic lesion (atypical hyperplasia): no wall thickening or nodule; noninvasive IPMT (adenoma and intraductal carcinoma): a nodule or wall thickening is present; and invasive IPMT with pancreatic parenchymal invasion: a mass with a heterogenous pattern or interruption of the pancreatic duct wall by the mass. RESULTS: The diagnostic accuracy rate for differentiating nonneoplastic lesion noninvasive IPMT, and invasive IPMT was 33% by ultrasonography, 38% by computed tomography, 77% by endoscopic ultrasonography, and 67% by intraductal ultrasonography. Sensitivity, specificity and accuracy rates for differentiating neoplastic and nonneoplastic IPMT by ultrasonography was 33%, 100%, 42%, by computed tomography 36%, 100%, 44%, by endoscopic ultrasonography 90%, 71%, 88%, by intraductal ultrasonography 94%, 29%, 84%, respectively. Sensitivity, specificity and accuracy rates for differentiating invasive and noninvasive IPMT by ultrasonography was 25%, 100%, 80%, by computed tomography 33%, 100%, 83%, by endoscopic ultrasonography 55%, 97%, 88%, by intraductal ultrasonography 56%, 91%, 84%, respectively. Diagnostic accuracy for invasive IPMT except minimally invasive cases by endoscopic ultrasonography and intraductal ultrasonography was 80%, based on the results of the examination which demonstrated a higher grade lesion. CONCLUSIONS: With these criteria, ultrasonography and computed tomography showed high specificity, but low sensitivity for the differential diagnosis of neoplastic/nonneoplastic and invasive/noninvasive IPMT. However, endoscopic ultrasonography and intraductal ultrasonography had high sensitivity and diagnostic accuracy for the differential diagnosis of neoplastic/nonneoplastic lesions. Combination of endoscopic ultrasonography and intraductal ultrasonography showed a high accuracy rate in the diagnosis of invasive IPMT. Thus endoscopic ultrasonography and intraductal ultrasonography contributed significantly to the choice of the treatment for IPMT. PMID- 11490850 TI - Surgical treatment of intraductal papillary-mucinous tumors of the pancreas. AB - BACKGROUND/AIMS: IPMT (Intraductal papillary-mucinous tumor of the pancreas) is increasingly recognized. The aim of this study was to investigate the appropriate surgical treatment for these tumors. METHODOLOGY: Between January 1981 and September 1998, 62 patients with IPMT underwent surgery. We retrospectively examined the clinicopathological features and surgical outcomes of the patients. RESULTS: The types of IPMT were as follows: hyperplasia (20); adenoma (31); and carcinoma, both invasive (5) and noninvasive (6). Lymph node metastasis was found in 36% of the carcinomas. The size of mural nodules was more than 3 mm in all adenoma or carcinoma cases, while the percentage of hyperplasia less than 3 mm was 75%. Intraoperative pancreatoscopy and annular array ultrasonography were very useful, because they detected 10 lesions that could not be found by preoperative examinations, such as computed tomography, endoscopic retrograde pancreatography, and endoscopic ultrasonography. All patients underwent surgical resection, including 10 pancreaticoduodenectomies (Whipple's procedure), 10 pylorus-preserving pancreaticoduodenectomies, 13 pancreatic head resections with segmental duodenectomies, 17 distal pancreatectomies, 9 segmental resections of the pancreas, 2 duodenum-preserving pancreatic head resections, and 1 total pancreatectomy. No operative or hospital death was observed. The postoperative survival rate at 5 years was 71.6% for carcinoma in IPMT. All of the cases with hyperplasia, adenoma and noninvasive carcinoma survived. Only two of the patients with invasive carcinoma died. CONCLUSIONS: IPMT had a favorable prognosis, as compared with pancreatic duct carcinoma. When selecting a surgical procedure for treating these tumors, it is important to confirm the tumor extent, as well as the diagnosis of invasion or noninvasion. In cases with invasion, radical resection is required. On the other hand, organ-function-preserving procedures should be selected for diseases without invasion. PMID- 11490851 TI - Problems in histological diagnosis of intraductal papillary-mucinous tumor (IPMT). AB - IPMTs (intraductal papillary-mucinous tumors) of the pancreas have been recognized as a distinct clinical entity. WHO used this term in most recent classification (1996). The present report reviews the WHO classification and recent descriptions of IPMT. Problems regarding the histological diagnosis and differential diagnosis are also discussed. In the WHO classification, IPMTs are classified into three categories: intraductal papillary-mucinous adenoma, intraductal papillary-mucinous tumor with moderate dysplasia and intraductal papillary-mucinous carcinoma. The classification is based on the tissue morphology, such as degree of dysplasia and pattern of proliferation. Some immunohistochemical and molecular markers have been reported for differential diagnosis and estimating the prognosis of IPMT. MUC1, Dpc-4, p53 and Ki-67. In making a differential diagnosis, mucinous cystic tumors are the most problematic. Communication with the pancreatic ducts, the presence of ovarian type stroma and capsular formation are key histological factors for a differential diagnosis between IPMTs and mucinous cystic tumors. The prognosis of IPMTs is favorable in general. However, once massive invasion has occurred, the prognosis is very poor, as in cases of ductal carcinoma. For further studies of IPMT, pathologists and clinicians involved in the diagnosis and treatment of IPMTs need to understand the concept of IPMTs and use the WHO classification. PMID- 11490852 TI - Topical nitrates and the higher doses of botulinum toxin for chronic anal fissure. AB - BACKGROUND/AIMS: Combined BT-A (botulinum toxin A) therapy and local application of nitrates can be more effective than BT-A alone for chronic anal fissure treatment, but so far the optimal dose of BT-A is not known. The aim of our study was to learn if BT-A doses higher than those used so far could change the outcome of fissure treatment. METHODOLOGY: We enrolled 14 consecutive patients suffering from idiopathic chronic anal fissure who did not respond to previous local treatment of nitric oxide donor and subsequent BT-A therapy (25 U of Botox). They were offered a local nitroglycerin treatment. In failure cases patients received the greater doses of BT-A (50 U of Botox). RESULTS: In all 11 patients with chronic anal fissure who applied nitroglycerin after BT-A injection, an effect on the internal anal sphincter relaxation was observed but fissure healing after topical nitroglycerin occurred only in 1 case. Of 13 patients with chronic anal fissure who received 50 U of BT-A no healing was reported in 6 cases. One male from this group received a greater dose (100 U of Botox) and then the fissure healed. CONCLUSIONS: The effect of topical nitrates on internal anal sphincter relaxation after botulinum toxin injection is not the last line for nonsurgical treatment of chronic anal fissure. Always we ought to consider using the next greater dose of BT-A before surgical treatment. PMID- 11490853 TI - Inferior mesocaval shunt for bleeding anorectal varices and portal vein thrombosis. AB - Intractable bleeding from anorectal varices is a serious and often misdiagnosed complication of portal hypertension and no agreement has been reached on which could be the optimal diagnostic and therapeutic strategy. Indeed, fatal outcome has been often reported resulting from delayed diagnosis and improper treatment. The case of a 67-year-old gentleman with life-threatening bleeding from anorectal varices who successfully underwent inferior mesocaval shunt is reported, and surgical technique for establishing a shunt between the inferior mesenteric vein and inferior vena cava is described. A review of other therapeutic options is presented and results are discussed and compared to those obtained with this novel form of treatment. In our experience, immediate control of recurrent bleeding from anorectal varices was obtained with inferior mesocaval shunt. Technical ease, promptness of action and effectiveness, low procedure-related morbidity are the main features of the shunt. With the introduction of new promising second-line treatment modalities to primary and metastatic liver tumors, like percutaneous radiofrequency thermal ablation, and improvement in outcome of portal vein thrombosis, the inferior mesocaval shunt may represent a sound alternative for patients who are ineligible for transjugular intrahepatic portosystemic shunt or presenting with clotted shunt. PMID- 11490854 TI - Surgical treatment for non-dilated biliary tract with pancreaticobiliary maljunction should include excision of the extrahepatic bile duct. AB - BACKGROUND/AIMS: The authors evaluated the surgical treatment for non-dilated biliary tract with pancreaticobiliary maljunction. METHODOLOGY: Sixty-nine patients with pancreaticobiliary maljunction were divided into 61 patients with the dilated biliary tract and 8 with the non-dilated biliary tract. The levels of amylase activity in the bile in the gallbladder and the bile duct, the incidence and severity of postoperative cholangitis, and cell proliferating activity of the biliary tract epithelium, examined the proliferating cell nuclear antigen labeling index (PCNALI), were examined. RESULTS: Of the 61 dilated type patients, 12 were of Ia, 1 was of Ib, 22 were of Ic, 25 were of IV-A, and 1 was of IV-B according to Todani's classification. Cancer was detected in 7 dilated type patients and in 3 non-dilated type patients. A high level of amylase activity was measured in the bile juice in both the gallbladder and bile duct in all of the patients with pancreaticobiliary maljunction. PCNALI of the biliary tract epithelium of the patients without cancer (dilated type: bile duct 11.4%, gallbladder 12.7%; non-dilated type: bile duct 5.9%, gallbladder 13.8%) was higher than that of the patients without pancreaticobiliary maljunction (bile duct 1.5%, gallbladder 1.4%). CONCLUSIONS: In a non-dilated type, as well as in a dilated type, a high level of amylase activity and increase of cell proliferative activity of the biliary tract epithelium were observed. Therefore, these results suggest that the extrahepatic bile duct should be prophylactically removed in patients with non-dilated type as well as in those with dilated type pancreaticobiliary maljunction. PMID- 11490855 TI - Acute biliary pancreatitis: staging and management. AB - BACKGROUND/AIMS: Acute biliary pancreatitis is a clinical entity with a high morbidity rate (15-50%) and mortality rate (20-35%). Early diagnosis has a primary importance for an appropriate treatment (75% of cases of idiopathic acute pancreatitis are of biliary origin). METHODOLOGY: Diagnosis of acute biliary pancreatitis in 78 patients was based on careful clinical and instrumental assessment: ultrasonography (76.9% of cases) and laboratory tests in 23.1% of cases. In our study we used the Ranson and APACHE II scores and 24 of the cases (30.7%) were classified as severe, while 54 (69.2%) were mild. All patients with severe acute biliary pancreatitis underwent emergency endoscopic retrograde cholangiopancreatography + endoscopic sphincterotomy (within 24-48 hours) followed by laparoscopic cholecystectomy (10 days). Patients with mild acute biliary pancreatitis underwent laparoscopic cholecystectomy associated with intraoperative cholangiography, within 10 days. RESULTS: In 19 patients with severe acute biliary pancreatitis operative endoscopy was curative. Subsequent laparoscopic cholecystectomy provoked subcutaneous emphysema only in one case but did not show any other serious morbidity. In the remaining 5 cases laparotomy was required because of necrosis, with a mortality rate of 60%. In all cases of mild acute biliary pancreatitis, laparoscopic cholecystectomy was successfully performed with a morbidity rate of 7.3%. Common bile duct stones were revealed with intraoperative cholangiography in 31.4% of the mild cases and in 75% of the severe cases. CONCLUSIONS: In conclusion acute biliary pancreatitis treatment is always surgical; in almost all severe cases it is performed with minimally invasive procedures (endoscopic retrograde cholangiopancreatography + endoscopic sphincterotomy with laparoscopic cholecystectomy < or = 10 days) if surgery is carried out within 24-48 hrs, as well in the mild cases (laparoscopic cholecystectomy + intraoperative cholangiography) when surgery is performed within 10 days. PMID- 11490856 TI - Comparison between pancreatoduodenectomy and hepatopancreatoduodenectomy for bile duct cancer. AB - BACKGROUND/AIMS: Pancreatoduodenectomy has been accepted as a standard operative procedure for distal bile duct cancer with low operative mortality. However, hepatopancreatoduodenectomy has not been accepted as a standard treatment modality for diffuse bile duct cancer. METHODOLOGY: From December 12, 1992 to December 15, 2000, 37 patients with the diagnosis of extrahepatic bile duct adenocarcinoma (cholangiocarcinoma) underwent pancreatoduodenectomy or hepatopancreatoduodenectomy, at the department of surgery, Ibaraki Prefectural Central Hospital and the Cancer Center. The differences in indications and results of both operative procedures were investigated retrospectively. RESULTS: Thirty-day operative mortality was 0% after either pancreatoduodenectomy or hepatopancreatoduodenectomy. One- to 5-year cumulative survival rates for the 24 patients after pancreatoduodenectomy were 76.3%, 41.5%, 41.5%, 41.5%, 41.5%, respectively. One- to 4-year cumulative survival rates for the 13 patients after hepatopancreatoduodenectomy were 48.0%, 32.0%, 32.0%, 16.0%, respectively. There were no statistically significant differences between cumulative survival rates after pancreatoduodenectomy and hepatopancreatoduodenectomy either in all the patients or in patients with UICC stage IV. CONCLUSIONS: Hepatopancreatoduodenectomy should be tried for patients with diffuse bile duct cancer, because only hepatopancreatoduodenectomy has the possibility of a cure at this time. PMID- 11490857 TI - Ventral dissection of replaced right hepatic artery during pancreatoduodenectomy. AB - A novel technique for dissecting a replaced right hepatic artery during pancreatoduodenectomy in patients with middle or lower bile duct carcinoma is presented. After skeletonizing the left, proper, common hepatic arteries and the portal vein, the replaced right hepatic artery is dissected from the ventro medial side of the hepatoduodenal ligament by severing the thin connective tissue behind the portal vein. Thus the hepatic arteries and the portal vein are completely isolated without the Kocher maneuver, leaving the cancer and the bile duct untouched en bloc with the surrounding lymph nodes and the pancreas head. The periaortic lymph nodes can also be resected en bloc with the main lesion. PMID- 11490858 TI - [Surgical therapy of female stress incontinence 2001. TVT (tension-free vaginal tape)--what else?]. PMID- 11490859 TI - The basic understanding and clinical results of tension-free vaginal tape for stress urinary incontinence. AB - Since 1994, over 100,000 surgical reconstructions of the female lower urinary tract have been performed worldwide using tension-free vaginal tape (TVT). This technique, which differs from traditional methods, is effective against symptoms of stress urinary incontinence and may be performed as an ambulatory procedure with no need of indwelling catheterization. The primary indication is genuine stress incontinence, but it is also effective as a secondary procedure for relapse after other anti-incontinence measures such as the Burch procedure. The TVT operation was developed according to study results on the mechanisms of urethral closure in women and has become the primary surgical procedure at several Scandinavian clinics for combating genuine stress incontinence in women. It is also useful in reducing symptoms after prolapse repair for combined prolapse and stress incontinence and in cases of mixed stress and urge ("sensory") incontinence. The small incisions and canals involved with the TVT technique minimize surgical trauma and enable it to be performed under local anesthesia. By the same token, it makes fairly small demands on postoperative care and attendant costs. Here we describe the instrumentation, procedure, and risks of this minimally invasive operative technique. PMID- 11490860 TI - [Sling-plasty in therapy of female urinary incontinence]. AB - Traditionally, women with type III stress incontinence (intrinsic sphincter deficiency) are treated with sling procedures, which have undergone multiple modifications during the last 90 years regarding surgical approach, sling course, and materials. The latest variation of the established sling concept is the tension-free vaginal tape (TVT) procedure. The choice of sling material influences the postoperative complication rate and reveals a conflict between unrestricted availability (alloplastic material) and optimal tissue compatibility (autologous material). Although valid information about the surgical outcome of sling procedures is rare, at least some evidence-based conclusions may be drawn from the meta-analysis of the published data: sling procedures and colposuspensions are more efficient and more durable than needle suspensions or anterior repairs in the treatment of female stress incontinence. The complication profile does not show a significant difference between slings and colposuspensions. The lack of long-term results for the TVT procedure precludes any definite assessment of this innovation. In any case, to meet the patient's interests, no surgical approach for correction of stress incontinence should be undertaken without complete diagnostic evaluation of the problem. PMID- 11490861 TI - [Submucous mucous membrane support in intrinsic sphincter insufficiency. A therapeutic approach in female stress incontinence]. AB - Urethral bulking agents are an attractive alternative for treating patients with intrinsic sphincter deficiency (ISD), i.e., type III urinary stress incontinence. These endoscopic techniques are minimally invasive and can be used in high-risk patients. As bulking agents, Teflon, autologous fat, collagen, silicone particles, and detachable microballoons have been the object of considerable clinical research. Teflon forms granulomas in the surrounding tissue and tends to migrate and is now considered obsolete. More recently, human collagen, autologous cartilage, Bioglass, and hyaluronic acid with dextranomer and polycarbon particles have come under clinical investigation, but long-term results are still lacking. The optimal bulking agent should be nondegradable and biologically inert and should not migrate or change its bulking capability. To date, there is no consensus on the best agent fulfilling these requirements, but encapsulated substances appear most promising. PMID- 11490862 TI - [Improvements in surgical therapy of incontinence--failed outcome!]. AB - The general advances made in minimal invasive surgery in the last 15 years has also led to the introduction of several new techniques for treating female incontinence. In the further development of bladder neck suspension according to Stamey-Pereyra, the use of miniature bone anchors received considerable support. Bladder neck suspension according to Stamey-Pereyra yields good initial results with a low complication rate but achieves permanent continence in only 40-71%. The anterior percutaneous implantation of miniature bone anchors with the attached suspension effects continence rates between 24% and 94%. Healing rates for transvaginal application of miniature bone anchors range from 52% to 100%. Reactions to foreign bodies are particularly common with synthetics but also occur with autologous materials. They are often associated with detrusor instability or sensory urge symptoms. Though these minimally invasive techniques can reduce the severity of stress incontinence, long-term healing is only achieved in about half the cases. The techniques described appear to be particularly unsuitable for treating grade III stress incontinence. The morbidity is unacceptable, especially when synthetic material is used in combination with bone anchors. Impaired vaginal wound healing often occurs in conjunction with irritative symptoms. PMID- 11490863 TI - [Colposuspension in treatment of female stress incontinence]. AB - Colposuspension has emerged as the "golden standard" for the surgical treatment of female urinary incontinence. With a lateral and tangential approach for the placement of the sutures, the rate of obstructed micturition and de novo urgency is low. Success rates of 85% for primary and 70-75% for recurrent incontinence can be achieved. In competition with the tension-free vaginal tape (TVT) procedure, colposuspension will remain the first choice for all laparotomies necessitated by other pathologies, in cases of paravaginal herniation as the cause of prolapse, and in women with unstable bladders caused by anatomical defects. The endoscopic approach has not yet gained general acceptance. PMID- 11490864 TI - [Surgical therapy of penis carcinoma]. AB - Primary therapy of penile cancer (carcinoma in situ/T1 tumors) consists of circumcision, microsurgical excision, application of 5-fluorouracil cream, radiation, or laser treatment. In cases of larger T1 tumors or T2 and distal T3 tumors, partial penectomy with a 2-cm margin of clearance is mandatory. Secondary therapy includes inguinal lymphadenectomy 4-6 weeks after primary treatment and antibiotic prophylaxis. Independent prognostic factors for the presence of lymph node metastases are T stage and grading. Only patients with noninvasive G1 or G2 tumors and nonpalpable inguinal lymph nodes are candidates for surveillance with careful follow-up. Inguinal lymphadenectomy is performed in a radical or modified (Catalona) manner. Sentinel biopsy (Cabanas) may regain importance with the use of gamma probes. Complication rates of inguinal lymphadenectomy correlate to the extent of the procedure and must be weighed against the possibility of cure with lymphadenectomy. In cases of inguinal lymph node metastasis, removal of the iliac lymph nodes (one- or two-step procedure) is necessary. PMID- 11490865 TI - [Radiotherapy of penis carcinoma]. AB - Penile cancer is rare. Thus, there are no therapeutic recommendations fulfilling the requirements of evidence-based medicine. The empirically based therapeutic approach consists of local excision, laser therapy, or radiotherapy with comparable local control rates. Radiation is delivered by external beam radiotherapy or as brachytherapy. After radiotherapy, 5-year survival rates of 66 92% and organ preservation in 55-84% are reported. Serious long-term sequelae are necrosis (3-23%) and urethral stenosis (6-45%) requiring surgery. In the adjuvant treatment of the locoregional lymph nodes, lymphadenectomy and radiotherapy of both inguinal regions are therapeutic options. Inguinal lymph node metastases may be irradiated pre- or postoperatively to reduce the local recurrence rates. In addition, palliative radiotherapy of the primary tumor, lymph node, or distant metastases is of use for incurable patients. New combined therapies, e.g., radiochemothermotherapy, are currently under clinical evaluation and may offer a curative and organ-preserving therapeutic option to patients with locally advanced tumors. PMID- 11490866 TI - [Radiochemotherapy of penis carcinoma]. AB - Some authors report successful use of radiochemotherapy in patients with penile cancer. The most promising chemotherapeutic agents in penile cancer are cisplatin, methotrexate, bleomycin, vinblastine, and vincristine. There are different protocols for the use of chemotherapeutic agents such as mono- or polychemotherapy in combination with radiotherapy. Operative treatment is still the primary approach in patients with penile cancer. However, in some patients with relevant co-morbidity who wish to receive organ-sparing therapy, radiochemotherapy may be applied when low-stage tumors (carcinoma in situ or T1) are present. There is no chemotherapeutic agent of choice to be recommended. The results of radiochemotherapy in patients with T2 tumors or higher are not satisfactory because local tumor control often cannot be achieved. PMID- 11490867 TI - [Fibromyxoid pseudosarcoma of the urinary bladder. Case report and review of the literature of a rare disease]. AB - Pseudosarcomatous fibromyxoidtumors (PSF) of the bladder are rare, benign tumors consisting of myofibroblastoid bladder wall proliferations of unknown etiology. Misinterpretation as a sarcoma is common and due to infiltrative growth and cellular pleomorphism and may leaed to false radical treatment. We present a 27 year old man with PSF presenting with microhematuria. Results of clinical, radiological and histopathological examinations and therapy are presented. A. review of the literature data of the to our knowledge 73 cases that have been reported in the literature is given. PMID- 11490868 TI - [Benign prostatic hyperplasia (BPH) syndrome. Surgical and interventional therapy options]. PMID- 11490869 TI - [Guideline for diagnosis and therapy of libido and erectioni disorders. Guidelines of the German Society of Urology]. PMID- 11490870 TI - [Reply of the authors to the letter by H. Palmtag. Intermittent urinary catheterization]. PMID- 11490871 TI - [Bioethical aspects of genetic studies in psychiatry]. PMID- 11490872 TI - [Changes in the recurrence rate of neurotic symptoms]. AB - The frequency of neurotic disorders' symptoms measured by symptom check-list in those admitted for specialist treatment, varied significantly even in a time-span of more than ten months (studies 1979-1999). PMID- 11490873 TI - [Main problems in psychoneuroimmunology. Part III]. AB - The paper presents the salutogenesis model of Antonovsky in the perspective of the psychoneuroimmunology. PMID- 11490874 TI - [The level of self-esteem in patients with anorexia nervosa]. AB - The aim of this work was to measure the self-acceptance level in patients with anorexia nervosa. The obtained results indicate that patients with anorexia nervosa have lower self-acceptance level in comparison with the control group. PMID- 11490875 TI - [Intergenerational family patterns in families with eating disorders and schizophrenia]. AB - The paper analyse the correlation between autonomy of patients with eating disorders and schizophrenia and autonomy of their parents. Triangulation within the 3-generations family system in both diagnostic groups is described. PMID- 11490876 TI - [To disclose or not to disclose the diagnosis of schizophrenia? A social constructionist approach]. AB - From a social constructionist perspective the author considers the advantages and disadvantages of using the word schizophrenia when talking with the patient and the family. PMID- 11490877 TI - [A map of delusional world according to Zdzislaw Bizon]. AB - The paper describes an instrument for presentation of content of delusions, which thanks to its synthetic graphic form, leads to improved systematization and understanding of delusional beliefs. PMID- 11490878 TI - [The way they see you, the way they perceive you: psychiatrists through the eyes of their patients]. AB - The article presents views of Polish psychiatric patients on their doctors' clothing styles at work and the way they address them. PMID- 11490880 TI - ["7 Minute Screening". Part I: a new tool in the diagnosis of Alzheimer's disease -- Polish translation and description of its clinical application]. AB - A polish translation and the description of the clinical use of the novel screening tool in Alzheimer's disease, a "7 Minute Screen", is presented. PMID- 11490879 TI - [Polish version of Temperament and Character Inventory (TCI): the analysis of reliability]. AB - The reliability of every main scale of the Polish version of Temperament and Character Inventory (TCI) was proven on the group of 144 students. PMID- 11490881 TI - ["7 Minute Screening. Part II: the evaluation of the test sensitivity and specificity as compared to Mini-Mental State examination]. AB - In the paper an evaluation of basic accuracy parameters (sensitivity and specificity) of the polish version of "7 Minute Screen" as a screening tool for Alzheimer's disease is presented as well as its comparison to Mini Mental State. PMID- 11490882 TI - [The involvement of the right hemisphere in affective manifestations]. AB - This paper concerns various aspects of the hemispheric contribution to the emotions formation. PMID- 11490883 TI - [Cost of depression: prospective analysis. Follow-up study in patients with recurrent depression]. AB - This article presents a comparison of costs of the first, the second, the third or next episode of depression, based on preliminary results of an observation clinical trial. PMID- 11490884 TI - [Mental status of subjects who 15 years ago (in childhood and/or puberty) were diagnosed with depressive disorders]. PMID- 11490885 TI - [The administration of St. John's wort (Hypericum perforatum) in the treatment of depression]. AB - Over the last years St. John's wort (Hypericum perforatum) has been extensively studied for its potential antidepressant activity. In Poland, it has not been approved yet as an antidepressant drug by the State Committee of Pharmaceutical Agents and Medical Materials Registration. However, since this agent is widely and commonly used (on a parapharmaceutical base), and also taking into account that possible hazard of such treatment and the risk of interactions with other drugs (which have been reported)--it seems reasonable to review and summarise the collected up-to-date data concerning these issues. PMID- 11490886 TI - [Phototherapy in non-seasonal affective disorders]. AB - This is a review of papers assessing effectiveness and tolerability of phototherapy in non-seasonal affective disorders. Phototherapy (therapy with bright, white light) is a well known method of treatment in seasonal affective disorder (SAD). The results in non-SAD are somewhat controversial. PMID- 11490887 TI - [Comparative studies of adverse effects in patients with refractory depression treated with amitryptyline, mianserin and unilateral ECT]. AB - The comparative studies of side effects in 180 patients with refractory depression treated with amitriptyline, mianserin and unilateral ECT were performed. PMID- 11490888 TI - [The effect of antidepressants on regional cerebral blood flow measured with 99m Tc HMPAO SPECT in depressive patients]. AB - The results of investigations of regional cerebral blood flow using 99mTc HMPAO SPECT in 19 depressed patients before treatment and after 6 weeks of pharmacotherapy are presented. PMID- 11490889 TI - [The influence of mood-normalizing agents on cognitive functions]. AB - The influence of lithium, carbamazepine or valproate treatment on the cognitive functions is presented. PMID- 11490890 TI - [Zinc homeostasis and glutamatergic system in the pathogenesis and treatment of depression]. AB - This article presents a review of the interactions between zinc and glutamate system in the pathophysiology and treatment of depression. PMID- 11490891 TI - [Case of depression in the course of pancreatic carcinoma]. AB - Author presents the diagnostic difficulties of a pancreas carcinoma case with depression. PMID- 11490892 TI - [Psychiatric disorders in thyroid and adrenal gland diseases]. AB - Clinical picture of psychiatric disturbances occurring in dysfunctions of thyroid or adrenal axes is presented. PMID- 11490893 TI - [Determination of Aeromonas spp. and Pseudomonas spp. in chlorinated water supply]. PMID- 11490894 TI - [Tuberculosis trends in the region of Abruzzo (1988-1998)]. PMID- 11490895 TI - [Disinfection procedures in digestive endoscopy. Experience at a large Roman hospital]. PMID- 11490896 TI - [Prevalence of tuberculin index in the school community of a health district. Considerations regarding currency of this procedure in public health]. PMID- 11490897 TI - [Fungal contamination of various cheeses from the south of Italy: hygienic sanitary considerations]. PMID- 11490898 TI - [Catering service and food microbiology]. PMID- 11490899 TI - [Limitations and/or prescriptions in health personnel. Study at a Veneto hospital]. PMID- 11490900 TI - [Environmental quality in operating rooms in Campania]. PMID- 11490901 TI - [Microbiological contents of superficial waters in a protected forest area]. PMID- 11490902 TI - [Trends of standards regarding health and safety at the workplace]. PMID- 11490903 TI - [Control of arterial pressure: aim for clinicians of any level of care]. PMID- 11490904 TI - [Round atelectasis]. AB - The objective of this study was to analyse the epidemiologic, clinical, radiological and functional characteristics as well as the evolutive pattern of a group of patients diagnosed of round atelectasis (RA). Patients with a radiological diagnosis of RA were retrospectively identified from January 1993 to January 1998. Cases with diagnosis not confirmed by high resolution computerized axial tomography (HRCAT) were excluded. A total of 29 patients were identified, with a mean age of 65 +/- 13 years (27 men and 2 women). At diagnosis 14 patients (34%) were smokers and 14 (49%) ex-smokers. Regarding occupation, 11 individuals (38%) had history of occupational exposure to asbestos. Regarding symptomatology, round atelectasis was a radiological finding in 15 patients (52%) and the most common symptom was chest pain (34%). The most common findings detected in the chest X-ray included pleural thickening (45%), pleural effusion (38%), nodular lesion (34%) and loss of volume (24%). The most common changes detected by HRCAT were pleural thickening (45%) and bronchovascular arch (55%). In two cases magnetic resonance (MR) was performed and in no case did this examination provided additional information for the diagnosis of RA. Functional respiratory examination did not identify and predominant pattern. During the follow-up period (2.2 years) 24 patients (83%) remained radiologically stabilized, one improved and the other four worsened (two due to enlargement and two due to increase in number). Chest X-ray is a good method for the presumptive diagnosis of RA. HRCAT is an excellent technique to confirm the diagnosis and rule out the presence of malignancy. MR provides no additional information in the study of RA. There is a frequent association between RA and asbestos exposure. PMID- 11490905 TI - [Control of arterial pressure: experience at a hospital monographic unit arterial hypertension]. AB - OBJECTIVE: To analyse the evolution of blood pressure control in patients referred to our centre for study, proportion of controlled hypertensive patients, changes in the characteristics of hypertensive patients and in therapeutic guidelines in the last twenty years. MATERIAL AND METHODS: A total of 4,103 patients were referred to study of hypertension. The analysed variables year by year included sex, age, weight, systolic blood pressure (SBP) and diastolic blood pressure (DBP). Cross sections at five-year intervals were performed to collect basal SBP and DBP and the evolution of BP in the following five years. RESULTS: From 1977 to 1997 the following changes were observed: basal mean age changed from 67 +/- 9 down to 56 +/- 10 years among males and from 68 +/- 8 down to 52 +/ 9 years among females; basal mean weight changed from 71 +/- 7 up to 81 +/- 8 kg among males and from 69 +/- 12 up to 73 +/- 13 years among females. Males showed a decrease in basal mean values of SBP/DBP from 176 +/- 25/112 +/- 20 mmHg down to 154 +/- 21/94 +/- 12 mmHg. In women, the corresponding values changed from 182 +/- 36/105 +/- 19 mmHg down to and 154 +/- 23/93 +/- 12 mmHg. In the early 1980's, 62% of patients received one or no antihypertensive drug. In the last few years such percentage has been 36% and 64% have been treated with two or more drugs. CONCLUSIONS: A decrease in age and an increase in weight of hypertensive patients referred to study have been observed in the last twenty years. Such patients show a less severe hypertension compared with referred patients in the early years. The use of new hypertensive drugs and their combined use are measures that contribute to improve the percentage of controlled hypertensive patients. PMID- 11490906 TI - [Medical training in the information society. Preparing legislation for health care revolution]. AB - The information society is continuously pushing to a rapid change und updating of laws and training and qualification programs from which medicine is not excluded. This paper summarises the norms and laws applicable to telemedicine with three basic principles involved: medical practice, data management, and communication technologies. Following the subsidiary principle, the applicable legislative levels are: European Union, National, Autonomous Community, Medical Professional Colleges, and Local Medical Colleges. Contradictory and little innovative issues appear in the results and discussion sections of the deontologic codes, the pressing demand on doctors' awareness and to provide them with technical skills. An exhortation follows to work out an informative-ethic code for the telemedicine practice. PMID- 11490907 TI - [Brucella hepatic abscess. Report of a case and review of the literature]. AB - Hepatic involvement in infections caused by Brucella organisms is common but the production of liver abscess is exceptional. Therefore, the most appropriate therapeutic approach is unclear. The use of antibiotics can be effective but in some cases abscess drainage is required. We report here a case of brucellar hepatic abscess and a review of fourteen cases reported in the literature. An emphasis is laid on the clinical, laboratory and radiological findings, treatment, and patient evolution. On the basis of the reported cases a therapeutic approach is proposed. PMID- 11490908 TI - [Indications and limitations of temporal artery biopsy in Horton arteritis]. PMID- 11490909 TI - [Indications and control of home oxygen therapy]. PMID- 11490910 TI - [Diagnostic strategies in interstitial pulmonary diseases]. PMID- 11490911 TI - [Clinical management of hyperprolactinemia]. PMID- 11490912 TI - [Applications of digestive echoendoscopy]. PMID- 11490913 TI - [Cutaneous and articular manifestations in a 52-year old male]. PMID- 11490914 TI - [Gingival hyperplasia in a patient with Wegener granulomatosis]. PMID- 11490915 TI - [Partial epilepsy refractory to medical treatment and occipital porencephalic lesion]. PMID- 11490916 TI - [Subcutaneous sarcoidosis with bracelet GA-67 uptake]. PMID- 11490917 TI - [Acute myositis ossificans]. PMID- 11490918 TI - [Gastric ulcer caused by cytomegalovirus in a patient with rheumatoid arthritis]. PMID- 11490919 TI - [Severe hypothyroidism secondary to interferon alpha]. PMID- 11490920 TI - [Extrahepatic manifestations of type I autoimmune hepatitis]. PMID- 11490921 TI - [Multiple fast growing pulmonary nodules as presentation form of large B cell primary lymphoma of the lung]. PMID- 11490922 TI - [Hepatitis B following autologous hematopoietic transplantation. Report of a case of inverse seroconversion]. PMID- 11490923 TI - Anatomic study of the tricuspid valve in children. AB - We performed an anatomic study of the right atrioventricular valve in children under one year of age using a conservative method of dissection of the heart valve. The main aspects studied were the number of cusps and their morphometric characteristics, such as the width of the base and the depth of the cusps. Other parameters studied were the number of papillary muscles, number of tendinous cords, and diameter of the fibrous ring and the last one were divided in three regions, anterior, posterior and septal for localization of cusps. Our results showed that the number of cusps varied from two to four. Three cusps was the commonest finding and the fourth cusp, if present, was classified as anterolateral in location. The anterior and septal cusps had bases bigger than those of the posterior and anterolateral cusps; the septal cusp was deeper than the others; and the number of tendinous cords was greater for the anterior and septal cusps than for the posterior and anterolateral cusps. In addition, the posterior region showed great variability: in 35.7% it was occupied by undeveloped valve tissue and the posterior valve in these cases was located anteriorly. PMID- 11490924 TI - Relations of the accessory nerve with the internal jugular vein: surgical implications in cervical lymph node clearances. AB - The position of the lateral branch of the accessory nerve in relation to the internal jugular vein is given variously by different authors. In surgery of the neck, and especially in conservative cervical lymph node clearances, the lateral branch of the accessory nerve is protected when it is situated lateral to the vein. However, when the nerve is medial to the vein there is a risk of damage to the internal jugular vein. A prospective preoperative study of 123 cervical lymph node clearances, as well as a dissection study of 5 fresh subjects, was carried out to determine the position of the lateral branch of the accessory nerve in relation to the internal jugular vein. The surgical study showed that the lateral branch of the nerve was anterior and lateral to the vein in 122 of the 123 clearances, while the cadaveric study found the nerve always anterior and lateral to the vein. Thus the risk of injuring the internal jugular vein during cervical lymph node clearances is very small. The differences observed by authors may be explained by collapse of the internal jugular vein observed during cadaveric dissections. PMID- 11490925 TI - Mediastinal lymphatic efferents from the diaphragm. AB - The prognosis of non small cell lung cancer (NSCLC) invading the diaphragm is poor, probably due to the richness of the lymphatic drainage of the diaphragm. The aim of this study was to determine mediastinal lymphatic efferents from the diaphragm. The diaphragms of 20 adult cadavers (77-104 years) were injected with a dye (modified Gerota's medium) to permit the lymph vessels to be catheterised and then dissected. Each stage of the dissection was described and photographed: 23 injections on the right and 25 on the left. Diaphragmatic lymph vessels passed to one of three lymph centres: posterior (paraaortic nodes, n = 16), anterior (juxtasternal nodes, n = 16) and mediastinal (visceral nodes, n = 16). From these lymph centres arose ascending lymph pathways: posteriorly to the thoracic duct (8/16), anteriorly along the internal thoracic vessels (10/16) and in the mediastinum to the peritracheobronchial nodes (6/10). Lymphatics from the diaphragm are abundant and drain towards mediastinal node lymph centres connecting to the blood stream via the thoracic duct. These lymph pathways are common with those of the pulmonary segments. Poor prognosis of NSCLC invading the diaphragm may be explained by the common lymphatic drainage of both the lung and diaphragm. PMID- 11490926 TI - Fusiform enlargement of the nasolacrimal canal: a rare anatomic variation. AB - To determine the standard width and shape of the nasolacrimal canal (NLC) in the coronal plane, a retrospective review of 950 coronal paranasal CT sections was undertaken. The coronal diameter of NLC was measured at its upper and lower ends and anomalies in its configuration noted. The mean diameters of the NLC were 4.8 +/- 1.2 mm and 4.7 +/- 1.1 mm on the right and left sides, respectively. There was no difference in the mean NLC diameter between the right and left sides, upper and lower portions or males and females. In two cases, an abnormal dilation of the NLC at its inferior portion was encountered. A marked enlargement of the NLC can occur in the absence of a neoplastic or obstructive process and may be detected as an isolated finding representing idiopathic or congenital enlargement. Segmental NLC dilatation is not always an indicator of pathology, particularly in asymptomatic cases, and should be considered a rare anatomic variation. PMID- 11490927 TI - Persistent primitive trigeminal artery associated with posterior communicating artery aneurysm and hypoplastic vertebral artery. AB - We report a patient who presented with headache due to diffuse subarachnoid hemorrhage. A four-vessel angiogram revealed a left posterior communicating artery aneurysm with fetal origin, persistent primitive trigeminal artery and hypoplastic vertebral artery on the same side. This association was considered coincidental and co-existence of a saccular aneurysm with an anatomical variation of intracranial vasculature is briefly discussed. PMID- 11490928 TI - The use of MR images to evaluate the risks associated with proximal locking of intramedullary tibial nails. AB - Vascular complications associated with locked intramedullary tibial nailing are infrequent but always serious, with a secondary amputation rate of 30%. These complications, based on an MR study of the anatomic relationships between the interlocking screws and the neurovascular or articular structures at the proximal end of the tibia, have been analysed. Two types of nails were used: one with anterior-posterior locking and a specially designed prototype with oblique locking. In particular this study emphasised the anatomic risk of injury to the popliteal structures when using anterior-posterior locking, as well as to the anterior tibial vessels and proximal tibiofibular joint when using oblique locking. As no locking method appears to be better than any other, the best is that associated with the lowest morbidity. Consequently, the use of frontal instead of anterior-posterior locking is recommended; the use of oblique locking, which this study has shown carries a much higher risk of complications, is strongly advised against. PMID- 11490929 TI - Anatomical basis of lumboscopy. AB - Lumboscopy provides an approach to the retroperitoneum for the majority of upper urinary tract operations. The technique involves a knowledge of specific anatomical landmarks that differ from those in classical open surgery, because of the inferior approach to the renal pedicle with the kidney initially mobilized in a ventral and cranial position. An anatomical study was conducted on five cadavers, using images from surgical operations, in order to describe the vascular elements of the retroperitoneal spaces as they are approached during lumboscopy, as well as to define specific anatomical landmarks. Identification of psoas major is crucial as it is then possible to locate the renal pedicle opposite and the area of the great vessels medially. During dissection of the renal pedicle the renal vein and its branches are the first elements to be approached. On the left side the distal portion of the renal vein can be confused with a reno-hemi-azygo-lumbar trunk, due to the initial ventral mobilization of the kidney stretching it. Because of the inferior view of the renal pedicle the reno-hemi-azygo-lumbar trunk can obscure part of the renal artery and may be confused with the renal vein. On the right side initial identification of the inferior vena cava on the medial aspect of psoas major facilitates identification of the gonadal, renal and suprarenal veins all located in the same plane. PMID- 11490930 TI - Arterial blood supply of the proximal humeral epiphysis. AB - The arterial blood supply of the proximal humeral epiphysis is known to derive mainly from the anterior humeral circumflex a. (ACA), but this description may minimize the role of the posterior circumflex humeral a. (PCA). The studies of Laing [9] and Gerber [3] emphasized the role of the ACA and of its branches, the ascending anterolateral artery and arcuate artery Thus, this description could not explain either the necrosis or the absence of necrosis in all the cases of fracture or dislocation of the glenohumeral joint. The evaluation of the risk of a vascular post-traumatic necrosis of the humeral head requires a knowledge of its arterial vascularization, and the aim of this study was to determine the respective areas of vascularization of both the humeral circumflex aa. 32 shoulders of adult cadavers were studied: the ACA and the PCA were injected with latex containing two differently colored fluids. The proximal humeral epiphysis was removed with the arteries. The extraosseous vessels and the coloration of the capsule were noted; then the epiphysis was sectioned in 5 mm horizontal scans, and the bone staining was studied in order to define the distribution of the arterial supplies. The origin of the ACA and PCA was common in only 10 cases. The mean diameters were: ACA 0.8 mm (0.3 to 2) and PCA 2.1 mm (1.5 to 4). The subchondral bone was colored in 29 specimens by the PCA, and by both the ACA in the cranial part and PCA in the caudal part in 3. The apex of the head was colored by the ACA in 7 cases, the PCA in 7 and both ACA and ACP in 1 case; the head was colored by the PCA in 17 and the ACA in 12 cases; the lesser tubercle by the ACA in 23, the PCA in 2 and both arteries in 7 cases; the greater tubercle by the PCA in 19, the ACA in 5 cases and both in 1 case; the intertubercular groove by the ACA in 29, the PCA in 1 and both arteries in 2 specimens. The arcuate a. was distributed along the metaphyseal side of the epiphyseal plate, and small branches crossed the plate to reach the epiphyseal side and give numerous anastomoses to the branches of the ACA or the PCA. The diameter of the ACA was constantly smaller than that of the PCA. Exclusive vascularization of the humeral head by the ACA was not confirmed. The roles of both the ACA and PCA remain important, and must be taken into account in evaluating the risk of necrosis after a fracture, by carefully considering the topography of the separation and the displacement of the different parts. PMID- 11490931 TI - Ideal intraspinal implantation site for the repair of ventral root avulsion after brachial plexus injury in humans. A preliminary anatomical study. AB - The advances made to date in root reimplantation for avulsion of the brachial plexus are modest considering that there are only reports from ten patients in the literature. However, the results are promising and should be applauded. The problem with reimplantation, in addition to the difficult surgical exposure, is to determine the ideal intraspinal implantation site for the graft. Given the non permissive substrate properties of the CNS white matter, the outcome of grafts elsewhere in the spinal cord need to be evaluated. An inappropriate implantation site might explain the still modest recovery in this challenging group of patients. Consequently the microscopic anatomy of the cervical spinal cord segments C5 to T1 was studied. The aims were to 1--determine the relationships between the ventral gray horn and the ventrolateral sulcus of the spinal cord, and 2--analyse the location and course of the motoneuron fibres passing through the white matter to reach the ventral root. On the basis of the present findings it is proposed that reimplantation of nerve grafts or rootlets should be performed directly through the ventral root exit zone in contact with the ventromedial region of the ventral gray horn at a depth of 2 mm, rather than into the white matter of the lateral aspect of the cord. Initially we used a posterior approach with total facetectomies. An anterior approach is now evaluated for current use. PMID- 11490932 TI - Unusual blood supply to the pancreas by a dorsal pancreatic artery. AB - A variation of the blood supply to the pancreas was observed in an 89-year-old female, in which the celiac trunk gave off four arteries: the hepatic, splenic, left gastric arteries and an additional dorsal pancreatic artery. One of the branches the dorsal pancreatic artery joined with the superior mesenteric artery to form a longitudinal anastomosis. The anterior and posterior pancreaticoduodenal arcades arose from branches of the superior pancreaticoduodenal and the dorsal pancreatic arteries. The inferior pancreaticoduodenal artery, a branch of the superior mesenteric artery, was missing. The majority of the pancreas was, therefore supplied by the dorsal pancreatic artery. The clinical implications of this finding are that the size, location and course of a dorsal pancreatic artery should be established given its central role in the blood supply to the pancreas observed in the present study. PMID- 11490933 TI - Accessory suprarenal gland: report of a case. AB - The suprarenal glands are normally located at the superomedial aspect of the kidneys. Accessory cortical masses are seen in approximately half of the newborn but usually disappear later. Several cases with accessory cortical tissues located near the main suprarenal glands have been reported but their usual locations have been rarely described. Here we report a case with accessory cortical tissue located on the right in the retrocrural space with compression symptoms. Such a lesion may be confused with lymphadenopathy radiologically. PMID- 11490934 TI - A case with dextrocardia, ventricular septal defect, persistent left superior vena cava and drainage of the great cardiac vein into the left internal thoracic vein. AB - A 17-month-old female patient was operated on for ventricular septal defect and pulmonary stenosis. During the operation several cardiac anomalies were observed; these were dextrocardia, ventricular septal defect, persistent left superior vena cava, drainage of the great cardiac vein into the left internal thoracic vein and a pericardial pouch. These anomalies have not previously been reported together. PMID- 11490935 TI - High origin and unusual suprarenal branch of a testicular artery. AB - In a 42 year-old male cadaver, the left testicular artery was found to originate from the anterior surface of the abdominal aorta at the level of origin of the left renal artery. It ran parallel and just inferior to the left renal artery and gave off a branch which supplied the left suprarenal gland. The course, relations and branching of this suprarenal branch differed from the very rare cases found in the literature. Awareness of the possible existence of such variations of the testicular arteries is of great importance during surgical procedures. PMID- 11490936 TI - [The year 1989. A turning point for pathology at the University of Leipzig?]. PMID- 11490937 TI - [Gerhard Domagk. A pathologist conquers bacterial infectious diseases]. PMID- 11490938 TI - [Morphological findings and medical insurance aspects in 371 exhumations]. AB - OBJECTIVES: The morphological findings in organ systems following exhumation, form the basis for answering a number of medical insurance issues. The aim of this study was to analyse the development of the number of exhumations performed and the medical insurance relevance over a 31-year-period. METHODS: A total of 371 exhumations, performed between 1967-1998 at the Institute of Pathology, Occupational Associations Hospital, Bochum, for medical insurance reasons were evaluated. RESULTS: The average number of days after burial was 74, ranging from 9 to 47.8. For the first third of the period investigated, the proportion of exhumations was 3.5% of all autopsy cases, for the second third this fell to about 0.4% and rose to 1.5% for the last third. In the first two-thirds, the main reasons for the exhumations were related to the grading and effects of pneumoconioses in connection with the cause of death. In the last third, asbestos associated diseases were mainly involved. In 99.2% of all cases, the autopsy results revealed important evidence for clarification of the medical insurance issues. The current catalogue of expectations listing the pathomorphological findings which can be expected after certain periods of internment, could be extended by our own results. PMID- 11490939 TI - ["Physiological" and "neoplastic" C-cell hyperplasia of the thyroid. Morphologically and biologically distinct entities?]. AB - C-cell hyperplasia (CCH) occurs regularly in the setting of type 2 multiple endocrine neoplasia (MEN2), either separately or in association with medullary thyroid carcinoma (MTC). It can also accompany sporadic MTC and appear without any tumour association. To test the practicability of the terms "physiologic" and "neoplastic", 18 cases with incidental sporadic, non-MTC associated CCH were investigated and the morphological patterns were described. We found CCH of various degrees, including so-called neoplastic CCH. In 16 of the 18 cases, a MEN2 setting could be ruled out by mutation analysis of the RET proto-oncogene. Morphologically, one can not distinguish with certainty between sporadic and hereditary or reactive and tumour-associated CCH. While MEN2-associated CCH can be regarded as true preneoplasia, sporadic CCH possesses variable biologic potential. The preneoplastic potential of sporadic CCH is still obscure. A pure morphological distinction between "physiologic" and "neoplastic" CCH regardless of the RET status should not be used. PMID- 11490940 TI - [Arachnoid cyst with consecutive brain stem atrophy, hypoglossal nerve paresis and tongue atrophy]. AB - We report the pathological anatomy of a patient aged 77 years at the time of death, who suffered from an arachnoid cyst situated near the right lower olive nucleus. This led to palsy of the hypoglossal nerve and partial gliosis of the brainstem with consecutive hemiatrophy of the tongue. This lesion did not seem to have any further effects onto clinical presentation. The patient died of a fulminant lung embolism and also presented an adenocarcinoma of the lung. PMID- 11490941 TI - [Disseminated infection by Histoplasma capsulatum with AIDS]. AB - Histoplasmosis is an illness which occurs very rarely in Europe and it is especially rare in Germany. A generalised infection with Histoplasma capulatum, a systemic mycosis of the mononuclear phagocyte system (MPS), occurs only in individuals with weakened immune systems. Within the framework of diagnostics, a pathologist can be confronted with histoplasmosis since there has been an increase in travel to and from endemic regions, as well as an increase in the number of diseases of the immune system. The presented case reports the histological intravital and post-mortem diagnostics of disseminated histoplasmosis in existing HIV-infection in the stage of manifest AIDS. PMID- 11490942 TI - [Pigmented pheochromocytoma. Case report with immunohistochemical and electron microscopic characterization]. AB - We present a case of pigmented adrenal paraganglioma in a 39-year-old female patient with associated neurofibromatosis type 1 (NF1). Histology showed features typical for phaeochromocytomas except for varying amounts of brown pigment within the cytoplasm of tumour cells, which proved to be melanin by histochemical and ultrastructural analysis. The occurrence of melanin is believed to reflect the origin of this neoplasm from multipotent cells of the neural crest. Pigmented phaeochromocytoma has to be taken in consideration in the differential diagnosis of pigmented neoplasms, especially in the adrenal gland, where it has to be discriminated from pigmented cortical adenoma (so-called black adenoma) and primary malignant melanoma. PMID- 11490943 TI - [Current standards for the diagnosis of breast carcinoma in routine practice]. AB - Every pathologist in active practice must be aware of and follow, set standards and recommendations for the diagnosis of breast carcinoma in order to ensure that the patient has the preconditions for an adequate therapy, regardless of the particular hospital setting. According to present-day knowledge, the standard obligatory examinations include conventional morphology (staging, typing and grading of the tumour) and the analysis of the steroid hormone receptor status (estrogen and progesterone). The immunohistochemical determination of tumour growth fraction with the Ki-67 antibody is economically justifiable, since it serves as an adjunct for histological/cytological tumour grading, a method that has a poor level of reproducibility. Since the introduction of herceptin immunotherapy, the determination HER-2/neu overexpression has gained a practical role and should now be included as an obligatory component of the diagnostic testing. Additional tumour markers are not components of routine diagnosis and these should be limited to clinical studies as long as their prognostic and therapeutic value have not been proven. PMID- 11490944 TI - [Baltic Bernstein amber. Its use in medicine, historical review and current prospects]. PMID- 11490945 TI - [War is the father of all things...(Heraclitus)]. PMID- 11490946 TI - [The UICC Telepathology Consultation Center. "Second opinion" over the Internet for pathologists]. PMID- 11490947 TI - [Abdominal compartment syndrome. 100 years later--is it not time to acknowledge the syndrome?]. PMID- 11490948 TI - [The abdominal compartment syndrome]. AB - The abdominal compartment syndrome (ACS) causes dysfunctions of various organs through a progressive unphysiologic increase of the intraabdominal pressure. While the primary ACS is a result of the underlying disease/injury, secondary ACS is caused by surgical interventions. In the severely injured patient intra- and/or retroperitoneal bleeding, edema of viscera due to systemic ischemia reperfusion injury following hemorrhagic shock, abdominal/pelvic packing, and laparotomy closure under tension lead to ACS. The clinical signs of ACS are a tense abdomen with a decreased abdominal wall compliance. Early signs of ACS are a rise in inspiratory pressure and oliguria. Manifest ACS results in anuria, respiratory failure, reduced intestinal perfusion, and low cardiac output syndrome. If untreated, patients die due to left ventricular failure. Diagnosis of ACS is made using the patient's history including the injury pattern, the symptoms, the time period between injury and the occurrence of organ dysfunctions, and the physiologic response to decompression. Frequent determinations of the bladder pressure represent the "golden standard" for early recognition of ACS. Decompressive laparotomy should be performed with a bladder pressure > or = 20 mmHg and rapidly restores impaired organ functions. In the case of a multiple injured patients in shock or with associated severe head injury decompressive laparotomy may even be carried out at a lower bladder pressure. The abdomen is left open. In most patients staged laparotomy is necessary. The final closure of the abdominal wall is carried out after the edema have resolved between day 6 and 8 after primary laparotomy. PMID- 11490949 TI - [Evaluation of subdural hemorrhage in infants after alleged minor trauma]. AB - QUESTIONING: Recently the discussion concerning the causes of infantile subdural bleedings (SDB) has become quite controversial. The wide-spread interpretation that most of these cases are the result of abuse, especially by the shaken-baby syndrome, was doubted, and the role of (even minor) accidental events was emphasized. METHODS: This situation should be analyzed basing on the official statistics of the causes of death in the city of Berlin (1978-1998) and the autopsy material of our institute (1978-1999). RESULTS: In this period, approximately 440.000 children lived their first year of life in our city. Only 80 violent deaths of infants (up to 1 year old) were recorded in the official statistics, including 27 deaths due to blunt forces, with 24 lethal head injuries as the main group. Only two cases were attributed to "falls under unclear conditions"; all other accidental cases were the results of traffic accidents or falls from a height. No death due to an undoubted minor fall was recorded, nor was any in our autopsy material. We investigated 10 cases of infantile SDB, all without skull fractures and gross brain injuries. Only 1 victim had a SDB of a significant volume; in all other cases only small amounts of blood were present in the subdural space. Bridging vein ruptures were directly demonstrated in 8 cases and were bilaterally in most instances; recently their detection has been simplified by postmortem x-ray using contrast material. All 10 cases were interpreted as typical acceleration-deceleration injuries (as in shaking), although only in 2 cases a confession of this procedure could be obtained. CONCLUSION: Comparing cases of accidental and non-accidental SDB in the literature, infantile SDB obviously cannot be looked at as a homogeneous entity: two quite different types should be kept separate: the patients suffering from an accidental SDB due to a minor fall mostly do not deteriorate immediately after the trauma, develop SDB of some volume, up to a space-occupying mass lesion, and have often a good prognosis. A lethal outcome is extremely uncommon; we have not observed a single case of an infantile lethal SDB resulting from such a minor injury for more than 20 years. The source of the SDB in those cases currently is unknown in most instances. The second group of infantile SDB includes the well known group of shaken-baby-syndrome: no adequate history, infants dead or nearly dead on clinical presentation, often a poor outcome if the event is survived, typically no significant volume of SDB, and--according to our experiences--in all cases BV ruptures. This combination of several BV ruptures with no significant subdural bleeding is not compatible with a supposition of a minor fall causing this. PMID- 11490950 TI - [Effect of graduate education on complication rate and costs of hip prosthesis implantation]. AB - PURPOSE: We studied the effect of surgical experience with regard to complications and costs after 3290 primary total hip arthroplasties (diagnosis: osteoarthritis). METHODS: We retrospectively analysed duration of clinical treatment and anaesthesia, blood loss, number of banked blood and perioperative complications (fractures of the shaft and acetabulum, dislocations, peripheral nerve injuries, thrombosis of the deep veins, embolism and deep wound infections) related to the experience of the surgeon (junior surgeons in comparison to the consultant) after 3290 primary THA's from 1974 to 1997. Cost analysis was performed by measurement of time and material expenditure. RESULTS: 39.2% of all operations were performed by junior surgeons. This number emphasizes the realisation of the order of education in our clinic. Both groups showed just few differences in number of duration of clinical treatment, banked blood, nerve leasions and embolism. We found statistically not significant increased rates of fractures and dislocations in the THA's performed by an unexperienced surgeon, however the rates decreased the last 14 years. Significantly higher rates of deep wound infection at the beginning decreased in the last 9 years to the level of experienced surgeons. Due to longer operation times, we calculated higher expenses of DM 85 per case in the group of junior surgeons. CONCLUSIONS: Using an educational program, carried out with an experienced surgeon, there is nearly no additional risk for the patient regarding the complication rates in THA's. However, there are higher financial expenses that have to be calculated. PMID- 11490951 TI - [Surgical treatment of injuries of the thoracolumbar transition--3: Follow-up examination. Results of a prospective multi-center study by the "Spinal" Study Group of the German Society of Trauma Surgery]. AB - OBJECTIVES: Prospective, multicenter study addressing late results after operative treatment of acute thoracolumbar spinal injuries. METHODS: 682 patients (T10-L2) were included and 372 (80%) were postoperatively followed for 2 1/4 years (4-61 months). RESULTS: Comparing the initially included patients (n = 682) with the study group (n = 372), no differences were observed and results were assumed to be representative. A C-type lesion or polytrauma significantly prolonged the hospital stay. The method of operative treatment did not affect the length of the rehabilitation period. Neurological improvement was observed in 3 out of 7 patients with complete, and in 44 out of 64 (69%) with incomplete lesion. The operative method did not affect the improvement rate. The physical capacity significantly decreased. After a mean of 1/2 year of disability only 71% returned to work. 48% returned to their preoperative physical level. The mean Hannover Spine Score was 68 points (preoperative 94, p < 0.001), indicating permanent impairment of function. The angle-stable internal fixator was superior in restoration of spinal alignment and best radiological results were noted after combined stabilization. Posterior stabilization lead to high re-kyphosing. No correlations between radiologic and clinical parameters were observed. CONCLUSIONS: All treatment methods under study were appropriate for achieving comparable clinical and functional outcome. The internal fixator is superior in restoration of the spinal alignment. Best radiological outcome is achieved by combined stabilization. Merely by direct reconstruction of the anterior column the postoperative re-kyphosing is prevented and a gain in segmental angle is achieved. PMID- 11490952 TI - [Hannover Fracture Scale '98--reevaluation and new prospects for an established score system]. AB - The Hannover Fracture Scale (HFS) was first introduced in 1983. As the treatment of open fractures has improved over the years, mainly due to new implants, the prognosis of open fractures has also changed. Thus a reevaluation of this limb salvage score has become necessary. DESIGN: Retrospectively all parameters of the HFS were evaluated in 182 open fractures of the upper and lower extremity treated in our institution between June '94 and June '96. Statistical means included the multivariant analysis, ROC analysis, calculation of sensitivity, specify and accuracy for the criterion of primary amputation. Finally the HFS'98 was established, which is characterized by eight domains (bone loss, skin injury, muscle injury, wound contamination, neurology, periost stripping, local and systemic circulation), a total score range from 0-22 points and a cut off point (amputation recommended) at a score of > or = 11. This score was then prospectively applied on another 87 open long bone fractures, treated in our institution between July '96 and Dec. '97. RESULTS: This validation of the HFS'98 revealed a sensitivity of 0.82 and a specify of 0.99. In comparison the NISSSA and the MESS presented a sensitivity and specify of 0.71, 0.99 and 0.82, 0.99 respectively based on data of the prospective study group. In addition the Gustilo classification for open fractures has been defined by certain score ranges of the HFS'98. CONCLUSION: In conclusion the HFS'98 has become a reliable extremity salvage score with a fairly high positive predictive value of 0.99, which is applicable for all open long bone fractures regardless of their location. PMID- 11490954 TI - [Diagnostic value of a clinical test for exclusion of fractures after acute ankle sprains. A prospective study for evaluating the "Ottowa Ankle Rules" in Germany]. AB - Previous studies have shown that clinical criteria can be used for determining the need for radiographs after spraining injuries of an ankle. Following the criteria of the "Ottawa Ankle Rules" (OAR) fractures could be ruled out with a sensitivity of 100% and a specificity of 50% while reducing radiographs by 28%. The aim of this study was to assess the "OAR" when applied in a German emergency department. Patients older than 18, who presented with blunt ankle trauma were examined by clinicians, then radiographs were ordered in all and the "OAR" were retrospectively applied. In 397 treated injuries 79 fractures were diagnosed and 5 patients had radiologically suspected fractures. Following to the "OAR" 58 were unnecessarly X-rayed and 5 fractures would not have been discovered, all of which were minor. Sensitivity using the "OAR" was 94% and specificity 17%. We found that 15% less radiographs can be ordered applying the "OAR". The "OAR" have the ability to rule out significant fractures at the ankle and midfoot. PMID- 11490953 TI - [Fractures involving the distal femoral epiphysis. Long-term outcome after completion of growth in primary surgical management]. AB - During a period of 12 years (1983-1994) we operated 13 patients with fractures involving the distal epiphyseal cartilage of the femur. All patients could be followed up in clinical and radiological examination following the completion of growth. By examination no patients had any complaints. The clinical examination revealed no limitation in range of motion of the hip or knee joint compared to the other side in any of the patients. 5 patients out of 13 (38%) were found to have a correct axial alignment with no axis deviation, shortening or lengthening of the leg. The other 8 patients (62%) presented the following one- or multidimensional deformities (compared to the contralateral, uninjured femur): 2 patients had varus deformities, 5 patients had valgus deformities, the axial malalignment in the frontal plane was always less than 10 degrees in all cases. 5 patients were found to have a shortened femur (between 8 and 19 mm), while an elongation of the primarily injured femur (10 and 15 mm) was established in 2 patients. On the basis of this study, we recommend that clinical and, if necessary, radiological and/or sonographic examinations are performed following the completion of growth in all patients with injuries involving the epiphyseal plate of the distal femur. PMID- 11490955 TI - [Changes in bone density of the femur after cement-free implantation of a modular hip prosthesis with a long shaft]. AB - Until now, no reports exist on the existence of femoral bone loss after hip arthroplasty using long-stem cementless prostheses in elderly patients. In a prospective evaluation the amount of bone loss (stress-shielding) after implantation of a long stem hip prosthesis in patients with femoral neck fractures (group A) or pertrochanteric femoral fractures (group B) was examined. Eleven patients (five from group A and six from group B) were treated with a long stem modular hip prosthesis (MHP). Change of bone mass was evaluated using quantitative computed tomographie (QCT) immediately following and at six months (group A and B) and twelve months (group A) after implantation of the prosthesis. Clinical results, expressed with the modified Harris Hip Score, and relative changes of bone mass were compared with mean periprosthetic bone mass of the femur after operation. After implantation of the MHP, the maximum decrease of mean femoral bone mass was 19.1% at six months and 20.2% at twelve months for group A and 29.5% at six months for group B. Bone loss in the proximal periprosthetic area was higher than in the distal part. There was no correlation between baseline values of bone mass and the amount of stress-shielding or clinical outcome. PMID- 11490956 TI - [Evaluation of cartilage changes within the scope of second-look arthroscopy 12 months after surgical reconstruction of anterior cruciate ligament rupture]. AB - The anterior cruciate ligament (ACL) rupture is among the most frequent injuries of the joints. Actually, there is no doubt regarding the necessity for surgical treatment. However, little is known about the progression of osteochondral damages and their distribution following reconstruction of the ACL. We treated 150 tears of the ACL either acute (median 7d) with augmented reinsertion (n = 41) and with autologous bone patella tendon graft (n = 24) or chronic (median 369d) with autologous bone patella tendon graft (n = 85). The study design was prospective. All osteochondral defects were documented in the six compartments. Second look arthroscopy and clinical examination were performed after twelve months in the course of removal of implants and possible changes of the osteochondral state were evaluated. Detoriation of osteochondral state was seen in 86 (57%) of 150 patients, while there was no statistical difference between the three groups. The time of operation had no influence on the osteochondral damage. There was a significant increase of chondral damages in the medial and retropatellar compartment after autologous bone patella tendon graft, while the changes after augmented reinsertion were diffuse. There were no significant differences in the clinical examination between the three groups regarding the Lysholm score and the functional and subjective outcome. However, postoperative instability was frequently observed (positive pivot-shift 25%, positive Lachmann test 57%). We suggest that postoperative instability should have led to the increased osteochondral damages. PMID- 11490957 TI - [Principles of intramedullary fracture stabilization. 1]. PMID- 11490958 TI - [High pressure injection injuries of the hand with surfacer]. AB - The most commonly injected materials in high-pressure injection injury to the hand include paint, automotive grease, solvents and diesel oil. High-pressure injection injuries are a frequently mismanaged problem, as they are often not recognized and treated early, which may result in a compromised function of the hand or even the amputation of a digit. Early diagnosis is an important factor, as well as the type and amount of injected material and the injection pressure at appliance. We present one case of high-pressure injection injury to the hand with the surfacer Caparol Accord. This report describes the principles of treatment and gives an overview of the literature about tissue injuries caused by components of different substances encountered in high-pressure injection injuries. PMID- 11490959 TI - [Therapeutic concept for preventing chronic phantom pain after traumatic brachial plexus lesion]. AB - We report on a 29-year-old motorcyclist, who had suffered a traumatic right side arm plexus lesion. The myelo-CT image showed a avulsion of the cervical roots C7/C8. Five days after the accident the patient complained of phantom pain in the right plegic arm and was presented to our acute pain service (APS). The patient complained of lancinating attacks of severe phantom pain in the right arm (visual analogue scale intensity of 80-100 pts.). The initial pain treatment was performed with PCA (piritramide), and because of the lancinating pain character carbamazepine treatment was introduced. The pain intensity increased under carbamazepine (VAS = 100 pts.), and after treatment with five cycles of salmon calcitonin infusion the pain intensity decreased (VAS = 10 pts). After withdrawal of the infusion therapy with salmon calcitonin the pain intensity increased up to VAS = 70 pts. TENS therapy five times per day showed no analgetic effect. We repeated the calcitonin-infusion therapy and after five i.v. cycles we continued with 200 I.U. salmon calcitonin intranasal per day. The initial phantompain intensity decreased (VAS = 40 pts.), but showed no long term analgesia. The additional psychological treatment with relaxation techniques (Jacobson/Bensen) showed the desired phantom pain relief. An interdisciplinary and multimodal cooperation between anesthesiologists, trauma surgeons, neurosurgeons and psychologists is needed for successful phantom pain treatment after traumatic brachial plexus lesion. Intravenous salmon calcitonin showed only short-term analgetic effect. PMID- 11490960 TI - [External femorotibial transfixation in femoral fracture with joint involvement in a child]. AB - Long bone fractures combined with joint injuries run a high risk of destabilising the articulations. Remaining joints incongruence can lead to early arthosis especially in cases of severe injuries or not achieved anatomical reduction. A number of osteosynthesis methods are available for anatomical repair of the articular facet. This report presents a seven years old boy with an open comminuted fracture of the distal femur and consecutive joint instability, treated with a Transfixation (Orthofix) of the knee joint. The functional results suggest this method as an alternative treatment. PMID- 11490961 TI - [Expert assessment of malpractice claims. Current status and prospects]. PMID- 11490962 TI - [Definition of rehabilitation relevant concepts. Basic principle for improving process quality]. PMID- 11490963 TI - [Comment on the contribution by C. Gerstenkorn et al. "Delayed diagnosis of odontoid fracture after whiplash trauma of the cervical spine"]. PMID- 11490964 TI - [Comments on the contribution by A. Kropfl et al. "Callus formation after reamed and unreamed femoral intramedullary nailing"]. PMID- 11490965 TI - [Comment on the contribution by A. Siegel et al. Fatal iliac vein injury caused by guide wire used during dynamic hip screw (DHS) implantation"]. PMID- 11490966 TI - [Is "unjustified admission" in trauma surgery preventable?]. PMID- 11490967 TI - [Rehabilitation]. PMID- 11490968 TI - [Rehabilitation nursing: thoughts on a specific role]. AB - This article presents some thoughts about nursing care rehabilitation. The specifications and the evolution of the place of practice, as well as the patients' needs, make the professional's role continue after initial learning. In relation to some particularities of the patient's rehabilitation needs, we suggest exploration of concepts allowing to develop the nurse's role in rehabilitation. The co-animation of the therapeutic project, health education and the relation to the patient will be the guiding principles to investigate our practice. While a break between the initial training and the practice of rehabilitation seems inevitable, it will be important to determine the nature of the continuous training which we can propose. PMID- 11490969 TI - [Common lumbago and returning to work: various thoughts about a complex problem]. AB - Non specific low back pain (NSLBP) is commonly a persistent or recurrent problem. In general the longer a worker is off work with NSLBP, the more disabling the condition becomes, the less successful any form of treatment, and the greater the probability of long term sickness absence. The obstacles to return to work are diverse. Scientific evidence shows that the development of chronic low back pain and disability depends more on individual and work-related psychosocial issues than on physical or clinical features. There is strong empirical evidence that treatment at the subacute stage (NSLBP lasting for approximately 5-12 weeks) is more effective at preventing chronic pain and disability than attempts to treat chronic intractable pain and disability once it is established. Active rehabilitation programmes should be interdisciplinary, adapted to the local socioeconomic context and include education toward overcoming fear avoidance beliefs and promoting self-care, some kind of active exercises, some behavioural principles of pain management, and some intervention at the workplace to help and assist the worker in early return to work. Some of these programs have produced desirable occupational outcomes when all the stakeholders in the disability problem (worker, employer, insurer, attending physician) worked together. PMID- 11490970 TI - [Evaluation of functional capacity]. AB - A functional capacity evaluation (FCE) is a detailed examination and evaluation that objectively measures the individual's current level of safe functional abilities, primarily within the context of the demands of competitive employment. It measures the ability to perform functional or work-related tasks and predicts the potential to sustain these tasks over a defined time-frame. Measurements of function from an FCE are compared to the physical demands of a job or other functional activities, and are used to make return-to-work/activity decisions, disability determinations, or to generate a rehabilitation plan. Scientific evidence on validity and reliability is limited so far. An FCE is time-consuming and cannot be recommended as a routine evaluation. PMID- 11490971 TI - [Role of psychotherapy at a rehabilitation clinic]. AB - The authors describe the psychopathology which is referred to the consultation liaison psychiatry in a rehabilitation center. They motive their choice for the cognitive behavioral approach of psychotherapy and present the more frequent psychotherapeutic interventions in this specific setting. PMID- 11490972 TI - [Can we still give a fibromyalgia diagnosis?]. AB - This article is an attempt at a critical analysis of the fibromyalgia concept. The author applies himself to describing how the profile of this syndrome, associating chronic widespread pain to allodynia, became more and more precise, until it obtained the status of disease. He emphasizes that the concept lost its initial meaning when the criteria, resulting from an ambitious scientific methodology, were used to establish a diagnosis on an individual scale. He comes to the conclusion that after a century of existence, fibromyalgia does not possess any specific quality that would distinguish it from other chronic widespread pain syndromes to make it a pure nosological entity. It is an artificial construct that adds nothing to the understanding of a pain phenomenon and allows no rational therapeutic approach. PMID- 11490973 TI - [Functional assessment of the hand in neurologic rehabilitation]. AB - Functional evaluation of hand is essential in neurorehabilitation because it provides crucial information about capacities/incapacities of patients during their daily activities. Then therapists can plan realistic aims. This evaluation is usually devoted to occupational therapists, who have different tests and scales to measure the functions of hand. The movements and gestures of hands are so various and complex that no test is better than another. Here we select a few specific tests to evaluate hand functions and we present their advantages and interest in neurorehabilitation. PMID- 11490974 TI - [Assessment of car driving skills in brain injured individuals]. AB - Independent driving is considered as very important for a full independence and social integration of disabled people. In these patient with disabilities or handicaps, the driving skills must be carefully assessed, especially in presence of a moderate dysexecutive syndrome. A precise neurological and neuropsychological examination is usually necessary to evaluate aptitudes for driving. Standard neuropsychological tests are often limited to value the brain injury patients. At the moment, assessment on driving simulators really improves the opinion of medical experts, as they give qualitative and quantitative data on reproducible conditions. In fact, in more than 80% of cases, the tests on driving simulators allow to determine if the patient is able to drive or no, with or without a specially adapted vehicle. However, in a small percentage of cases, the evaluation of open-road driving by a driving instructor is necessary, proving that driving simulators will never take over from reality. PMID- 11490975 TI - [The aphasia group of Sion: 15 years of experience]. AB - The overall plan for this article reflects the author's experience of a full rehabilitation program for speech therapy in group. The organisation, the procedures, and the goals of an aphasia group are presented. The efficacy of such a training is discussed based on the controversial literature. The main current thoughts about this specific treatment as well as theoretical rationale for treatment processes are explained. PMID- 11490976 TI - [Spinal cord injury: potential for neurologic and functional recovery]. AB - Since antiquity, spinal cord injury was recognised mortal. At the beginning of the 19th century, the situation had hardly changed and the mortality rate remained high. Since the Second World War the care of the spinal man evolved considerably. Over the last past years, promising experiments on the neurological recovery in animals were achieved. While waiting for their application to the human being, global rehabilitation in specialised centres offers to spinal cord injured patient the possibility of functional recovery with social and professional reinsertion. PMID- 11490977 TI - [What to do in a case of acute ear pain?]. PMID- 11490978 TI - [Current indications for the treatment with botulin toxin]. AB - Botulinum toxin is more and more frequently used as a therapeutic agent. The toxin blocks selectively and reversibly the neuromuscular junction, causing a muscle relaxation. Indications are mainly muscular hypercontraction, such as dystonia, blepharospasm, focal spasticity, strabismus or tics. The range of action extend to focal hyperhydrosis, palmar, axillary or plantar. It seems now that some painful syndrome such as migraine or tension headache may benefit from toxin injections. Esthetic indications constitute an extension to the pure medical indications. PMID- 11490979 TI - [Intensive Psychiatric Support at the Setting in Lausanne: a pilot project]. AB - While the hospital remains an important element of the psychiatric health-care system, the fact that it is always the best place to treat acute psychotic episodes is still debated. After a brief review of the literature relative to the main existing community care models, the authors describe the development in the Department Universitaire de Psychiatrie Adulte (DUPA), of an alternative to hospitalisation for patient going through a severe acute psychiatric episode. They present three clinical situations and the aims of the research project, which will follow this pilot phase. PMID- 11490981 TI - [Communication and confidentiality in the management and follow-up of cases of child abuse --several principles]. PMID- 11490980 TI - [The health professional's approach to chronic lumbago: clinical experience at the Spinal Cord Unit at the orthopedic hospital]. AB - The treatment of back pain patients refers to the biopsychosocial model of care. This model includes illness in patient's personal and relational life. In this context, it is not only the physical symptom of the patient which is focused but also his psychological distress often hidden by algic complain. Clinical interviews conducted with back pain patients have highlighted psychosocial aspects able to influence the relationship between health care user and provider. Taking account of psychosocial aspects implies an interdisciplinary approach that identify and assesses patients' needs through adequate tools. As a result, the different health care providers implied with back pain patients have to collaborate in a structured network. PMID- 11490982 TI - Blue 'spot' on the nose. PMID- 11490983 TI - Prevention for the older woman. A practical guide to assessing physical and cognitive function. AB - Life expectancy of women is greater than that of men at every age. Thus, an older woman is more likely than a man to be living without a spouse and living alone. It is important in caring for an older woman to establish how well she is functioning physically and cognitively and in what activities she may need assistance. Given the high prevalence of dementia with advanced age, screening for dementia should be considered in women over age 75. Although their risk of suicide is lower, older women are more likely to be depressed than older men, regardless of race, ethnic background, or economic status. Late-life depression is treatable, and screening should be considered in the presence of a major life change, recent stroke, or MI. PMID- 11490984 TI - New guidelines on high cholesterol. PMID- 11490985 TI - Preoperative assessment. Primary care work-up to identify surgical risks. AB - For many older patients, surgery is an inevitable component of healthcare. Thorough and effective preoperative patient evaluation is imperative because comorbidities, polypharmacy, functional problems, and socio-economic issues all can affect perioperative decisions and complicate recovery. A methodical assessment includes a history and physical exam, review of prescription and nonprescription medications, evaluation of major organ systems, and discussion with the patient of advance directives. Other major considerations include functional status, the risk level of the surgery to be performed, expected blood loss, and anesthesia duration. The primary care physician must synthesize and relay this patient data to the surgical team so that appropriate measures can be taken to minimize perioperative risks and promote uncomplicated recovery. PMID- 11490986 TI - Suicide in late life. Identifying and managing at-risk older patients. AB - The patient in this case displayed multiple risk factors for suicide, including depression, loneliness following recent widowhood, use of alcohol, and distribution of possessions to other people. In retrospect, it was found that he was stockpiling old medications and refused to comply with his plan of care. The patient's outcome was tragic despite numerous attempts to provide a support system to meet his needs. Although he appeared to improve with therapy, the patient may have had a suicidal agenda that he managed to hide from everyone, including his family and multiple healthcare professionals. The patient's daughter was offered bereavement counseling services, which she appreciated. PMID- 11490987 TI - Updated NCEP guidelines set new decision points for managing dyslipidemia. PMID- 11490988 TI - Vector-borne infections in the tropics and health policy issues in the twenty first century. AB - Over the past 2 decades scientific advances and evolving strategies have significantly contributed to improved tools for control of vector-borne infections. These are: diagnostics--rapid assessment methods, non-invasive or minimally so yet sensitive and specific; new chemotherapeutics; pyrethroid insecticides and biological insecticidal products; refined strategies, such as combination therapy, rotation of insecticides for resistance management, community-directed treatment, standardized monitoring and evaluation to define programme progress; better epidemiological knowledge through improved identification of parasites and vectors; GIS, remote sensing and climate models which provide tools for epidemic prediction, planning control programmes and permit effective policy analysis; greater involvement of NGDOs (non-governmental development organizations) and CSOs (civil society organizations) in control; advent of donation programmes which involve community-based or directed mass drug distribution. Future problems could be: (1) the over-emphasis on inflexible financing by the insistence of donors on SWAps (sector-wide investment), (2) the over-reliance on pyrethroid pesticides, (3) the over-expectation that basic research will provide new drugs and vaccines for resource-poor settings in the necessary time scales, and (4) the failure to recognize that biological processes have an inherent capacity for change which outstrips the capacity of health services to respond. Malaria is a paradigm of an 'emerging disease'. (5) The challenge of implementing a 'vertical' approach to disease control within national health programmes, in the face of significant donor opposition to such programmes is a challenge even when such approaches will secure a 'public good'. PMID- 11490989 TI - The increase in risk factors for leishmaniasis worldwide. AB - Economic development leads to changing interactions between humans and their physical and biological environment. Worldwide patterns of human settlement in urban areas have led in developing countries to a rapid growth of mega-cities where facilities for housing, drinking-water and sanitation are inadequate, thus creating opportunities for the transmission of communicable diseases such as leishmaniasis. Increasing risk factors are making leishmaniasis a growing public health concern for many countries around the world. Certain risk factors are new, while others previously known are becoming more significant. While some risk factors are related to a specific eco-epidemiological entity, others affect all forms of leishmaniasis. Risk factors are reviewed here entity by entity. PMID- 11490990 TI - Screening for severe anaemia in pregnancy in Kenya, using pallor examination and self-reported morbidity. AB - Severe anaemia in pregnancy is an important preventable cause of maternal and perinatal morbidity and mortality. Different methods of screening for severe anaemia in pregnancy were evaluated in a 2-phased study conducted in Kilifi, Kenya. In phase 1 (in 1994/95), pallor testing was evaluated alone and in addition to raised respiratory/pulse rates: 1787 pregnant women were examined by one of 2 midwives. Sensitivities for detecting severe anaemia (haemoglobin < 7 g/dL) were 62% and 69% and specificities 87% and 77%, respectively for each of the midwives. Addition of high pulse rate increased sensitivity to 77% and 81%, but specificity reduced to 60% and 51%, respectively. In phase 2, following qualitative in-depth work, a screening questionnaire was developed. An algorithm based on screening questions had 80% sensitivity and 40% specificity. Midwife pallor-assessment was conducted following the screening questionnaire. In this phase (conducted in 1997), the midwife performed very highly in detecting severe anaemia, achieving sensitivity of 84% and specificity of 92%. Spending a few minutes asking women questions may have improved the ability to interpret pallor findings. This study demonstrates the value of pallor testing and raises alternative approaches to improving it. PMID- 11490991 TI - Impact of installation of a water pump on schistosomiasis transmission in a focus in Cameroon. PMID- 11490992 TI - Epidemic dengue transmission in southern Sumatra, Indonesia. AB - An outbreak of dengue fever (DF), dengue haemorrhagic fever (DHF), and dengue shock syndrome (DSS) in the city of Palembang, south Sumatra, Indonesia was investigated to (i) validate epidemic occurrence, (ii) confirm dengue virus aetiology and associated serotype(s), (iii) provide a demonstrable measure of community impact, and (iv) identify causative relationship (if any) with climatic El Nino Southern Oscillation (ENSO) influences. Trend analysis based on a 6-year retrospective review of hospital records demonstrates a 3-fold increase in clinical cases for the outbreak period (January-April 1998), relative to historical records. In the 2 hospitals surveyed, the monthly mean number of outbreak-related dengue cases over 4 months was 833 (range 650-995 cases/month); the mean monthly value for the previous 72 months was 107 (range 14-779 cases/month). An apparent trend in epidemic transmission was observed, evolving from a 5-year cyclic phenomenon to an annual occurrence, often indistinguishable from one year to the next. The proportional distribution of clinical outbreak cases into DF, DHF and DSS diagnostic categories was 24%, 66%, and 10%, respectively. The population aged 10-19 years accounted for the largest (35%) proportion of hospitalized DHF cases, followed by children aged 5-9 years (25%) and children aged 4 years (16%). Serum samples obtained during acute illness from 221 hospitalized patients were examined using serology, RT-PCR, and virus isolation in cell culture: 59% of samples had laboratory evidence of a dengue infection. All 4 dengue virus serotypes (DEN 1-4) were identified in epidemic circulation, with DEN 3 predominating (43%). DEN 1 was the principal serotype associated with less severe dengue illness, suggesting that virulence may be, in part, a function of infecting serotype. The climatic influence of ENSO on rainfall and temperature in the months leading up to and during the outbreak was dramatic, and is likely to contribute to favourable outbreak conditions. PMID- 11490993 TI - Validity of Lot Quality Assurance Sampling to optimize falciparum malaria surveys in low-transmission areas. AB - To control the reappearance of malaria in the Madagascan highlands, indoor house spraying of DDT was conducted from 1993 until 1998. Before the end of the insecticide-spraying programme, a surveillance system was set up to allow rapid identification of new malaria epidemics. When the number of suspected clinical malaria cases notified to the surveillance system exceeds a predetermined threshold, a parasitological survey is carried out in the community to confirm whether or not transmission of falciparum malaria is increasing. Owing to the low specificity of the surveillance system, this confirmation stage is essential to guide the activities of the control programme. For this purpose, Lot Quality Assurance Sampling (LQAS), which usually requires smaller sample sizes, seemed to be a valuable alternative to conventional survey methods. In parallel to a conventional study of Plasmodium falciparum prevalence carried out in 1998, we investigated the ability of LQAS to rapidly classify zones according to a predetermined prevalence level. Two prevalence thresholds (5% and 15%) were tested using various sampling plans. A plan (36, 2), meaning that at least 2 individuals found to be positive among a random sample of 36, enabled us to classify a community correctly with a sensitivity of 100% and a specificity of 94%. LQAS is an effective tool for rapid assessment of falciparum malaria prevalence when monitoring malaria transmission. PMID- 11490994 TI - The life-cycle of Leishmania infantum MON-77 in the Priorat (Catalonia, Spain) involves humans, dogs and sandflies; also literature review of distribution and hosts of L. infantum zymodemes in the Old World. PMID- 11490995 TI - Soil-transmitted helminth infections at very high altitude in Bolivia. AB - A cross-sectional study of soil-transmitted helminthiases in the Northern Bolivian Altiplano was carried out over the 6-year period 1992-97. Prevalences, intensities and associations were analysed from coprological results obtained in 31 surveys (28 in schools and 3 in individuals of all age-groups) performed in 24 Aymara communities located between the city of La Paz and Lake Titicaca, at an altitude of 3800-4200 m. Ascaris lumbricoides and Trichuris trichiura were detected, with local prevalences in the range 1.2-28.0% and 0.0-24.0%, respectively. Significant differences in prevalence rates of trichuriasis were detected, with highest prevalences in male schoolchildren and in subjects aged > 40 years. The global intensity ranged from 24 to 86,544 eggs per gram of faeces (epg) and from 24 to 4560 epg for ascariasis and trichuriasis, respectively. Higher intensities were noted in girls. A. lumbricoides egg counts were statistically significantly higher in the 5-8-years age-group. A positive association between A. lumbricoides and T. trichiura infections was detected. The proportion of heavy infections for A. lumbricoides was 0.1% and 1.0% in the school and community surveys, respectively. No heavy infection for T. trichiura was detected. The very high altitude and its severe environmental conditions may determine the relatively low prevalences and intensities in this area. PMID- 11490996 TI - Migration and dispersal of lymphatic filariasis in Papua New Guinea. PMID- 11490997 TI - A recombinant antigen-based IgG4 ELISA for the specific and sensitive detection of Brugia malayi infection. AB - An IgG4 ELISA based on a novel recombinant antigen was evaluated for detection of Brugia malayi infection, using 2487 sera from various institutions: 2031 samples from Universiti Sains Malaysia, 276 blinded sera from 2 other institutions in Malaysia, 140 blinded sera from India and 40 blinded sera from Thailand. These sera were from various groups of individuals, i.e., microfilaraemics, chronic patients, endemic normals, non-endemic normals and individuals with other parasitic and bacterial infections. Based on a cut-off optical density reading of 0.300, the IgG4 ELISA demonstrated specificity rates of 95.6-100%, sensitivity rates of 96-100%, positive predictive values of 75-100% and negative predictive values of 98.9-100%. These evaluation studies demonstrated the high specificity and sensitivity of this test for the detection of active B. malayi infection. Thus, the IgG4 ELISA would be very useful as a tool in diagnosis and in elimination programmes for brugian filariasis. PMID- 11490998 TI - Immunodiagnostic differentiation of alveolar and cystic echinococcosis using ELISA test with 18-kDa antigen extracted from Echinococcus protoscoleces. AB - Immunoblotting analysis was carried out using crude extracts of Echinococccus multilocularis and E. granulosus. A total of 214 serum samples were examined for the detection of antibody responses against 18-kDa polypeptides (Eg18 and Em18). Of 44 sera from patients with alveolar echinococcosis (AE) 91% were positive against both Eg18 and Em18, compared with 10% and 13%, respectively, of 70 cystic echinococcosis (CE) samples and 13% and 17% of 29 cysticercosis samples. A relatively purified 18-kDa antigen was extracted from E. granulosus protoscoleces and an 18-kDa-ELISA test which is simple, fast and highly sensitive and specific has been established. A comparative ELISA analysis, using the purified 18-kDa antigen and pooled AE and CE serum, indicated that there are significant differences in antibody levels between AE and CE sera. The maximum ratio for the evaluation of AE to CE was 2.26. The 18-kDa-ELISA was positive for 91% (40/44) and 11% (8/70) of AE and CE sera, respectively, and there was no cross-reactivity with cysticercosis sera or healthy controls. The overall diagnostic values of 18 kDa-ELISA for AE sera were sensitivity 91%, specificity 94%, positive predictive value 83% and negative predictive value 97%. We conclude that (i) the 18-kDa antigenic component may not be species specific but (ii) levels of antibody to the 18-kDa antigen are significantly different between AE and CE sera. Therefore, the 18-kDa antigen can be a reliable serological marker for differentiation of AE from CE. PMID- 11490999 TI - Serodiagnosis of ascariasis with specific IgG4 antibody and its use in an epidemiological study. AB - In an earlier study Ascaris-specific IgG4 antibody was found to be elevated in cases of ascariasis. However, the usefulness of the elevated levels of this antibody in Ascaris infection as a diagnostic marker has not been well established. In India, in early 1999, blood samples of 83 cases of Ascaris infection, 35 cases of other nematode infection and 53 control subjects (without any helminth infection) were tested for anti-Ascaris IgG4 by ELISA. Further anti Ascaris IgG4 levels in the blood of Ascaris-infected patients were determined, after eradication of the worms with drugs, at regular intervals to ascertain the duration of elevation of titre of the serological marker following initial infection. This information would indicate the sensitivity of the test as a diagnostic marker for recent infection. Blood samples of 422 rural people were also tested for anti-Ascaris IgG4 titre to ascertain the prevalence of ascariasis in the community. High levels of anti-Ascaris IgG4 antibody (OD 1.246 +/- 0.212) were found in all the 83 Ascaris-infected subjects compared to controls (OD 0.158 +/- 0.047). Anti-Ascaris IgG4 antibody levels of other nematode-infected subjects were comparable to the controls. Anthelmintic treatment of 8 Ascaris-infected subjects caused sequential fall of IgG4 level in their blood, and its titre reached control level within 6 months of deworming. Of 422 individuals from the rural community 229 (54.3%) had significantly high levels of specific IgG4 antibody against Ascaris excretory-secretory antigen, suggesting that they were infested with Ascaris. Thus, this study demonstrated that anti-Ascaris IgG4 antibody is a very sensitive and specific marker for the diagnosis of Ascaris infection. Utilizing this test, a significant number of a rural population could be diagnosed with Ascaris infection in West Bengal, India. PMID- 11491000 TI - Detection of malaria parasites in paraffin-embedded spleen and placental tissues by nested PCR. PMID- 11491001 TI - Natural progression of Oesophagostomum bifurcum pathology and infection in a rural community of northern Ghana. AB - An estimated 250,000 people in northern Ghana and Togo are infected with the intestinal helminth parasite Oesophagostomum bifurcum, as detected by stool cultures. Clinical disease caused by O. bifurcum is responsible for about 50 cases per year at the region's central hospital, and presents as painful abdominal masses: inflammatory colonic nodules containing live juvenile stages of the helminth. In individuals living in villages highly endemic with O. bifurcum infection, colonic pathology visible by ultrasound is also highly prevalent. These nodules also contain O. bifurcum juvenile worms but are apparently asymptomatic. Thus, O. bifurcum infection and asymptomatic colonic pathology are highly prevalent within this area, but clinical disease is relatively uncommon. The natural evolution and regression of the colonic pathology in an endemic community in northern Ghana and its distribution within the population is described. Of the 299 individuals in the study group, 28% had colonic pathology at recruitment in the late-rainy season, which decreased with a half-life of 3-4 months during the dry season. Of those negative at recruitment, 28% developed nodules during the year, the majority appearing at the end of the subsequent rainy season. Children tended to have a higher prevalence and intensity of ultrasound-visible pathology compared to adults. Almost half (49%) of the study group had colonic nodules at least once during the year, and 2% of these individuals presented with clinical disease to the local hospital during the mid rainy season. PMID- 11491002 TI - Presence of Schistosoma mansoni eggs in the cervix uteri of women in Mwanga District, Tanzania. PMID- 11491003 TI - Zoonotic infections as causes of hospitalization among febrile Bedouin patients in southern Israel. AB - A prospective 12-month study was conducted throughout 1998 to determine the frequency of selected bacterial zoonoses as causes of fever among hospitalized Bedouins in southern Israel. One or more zoonoses were diagnosed in 30 (27%) of 110 patients admitted with fever. Brucellosis was diagnosed in 9 (8%), rickettsial infections in 20 (18%), and ehrlichiosis in 2 (2%), one of whom had also evidence of rickettsial spotted fever infection. None of the patients was diagnosed with Q fever. Compared with patients without zoonoses, patients with zoonoses were younger (P = 0.01), fewer of them had underlying conditions (P < 0.02), they had a longer febrile period prior to hospitalization (P = 0.04), a significantly higher proportion had arthralgia (P = 0.02), rash (P = 0.03), and splenomegaly (P = 0.04) and a lower proportion had pathological findings on chest auscultation (P < 0.01). Patients with zoonoses were found to have more commonly anaemia (P = 0.03) and leucopenia (P = 0.02) compared to the rest of the study population. Of the 30 patients with zoonoses 60% were misdiagnosed and only 57% received adequate antibiotic treatment. Zoonotic infections are a common cause of fever in adult Bedouins living in southern Israel. Because of the non-specific features of these diseases they are often misdiagnosed. Blood cultures and multiple serological tests should be used in the investigation of fever in such patients and tetracycline should be considered for initial empirical treatment. PMID- 11491004 TI - Asymptomatic intestinal microsporidiosis in Thai orphans and child-care workers. PMID- 11491005 TI - Dengue virus infection during infancy. PMID- 11491006 TI - Can pretreatment screening for dhps and dhfr point mutations in Plasmodium falciparum infections be used to predict sulfadoxine-pyrimethamine treatment failure? AB - This study examines the relationship between malaria treatment failure after sulfadoxine-pyrimethamine (S-P) chemotherapy and presence of mutations in the Plasmodium falciparum dihydropteroate synthase (dhps) and dihydrofolate reductase (dhfr) genes (associated with resistance in vitro to S and P) before treatment. In Kenya, 38 malaria patients in a holoendemic area, and 21 in an epidemic area, participated in the trial in 1997-98. In the 2 areas, drug failure occurred in 76% and 75% of cases where any mutation in dhfr was seen (positive predictive values 76% and 75%: P = 0.003 and 0.008) and an identical association was seen with dhfr Asn-108. In the holoendemic area all occurrences of > or = 2 mutations in dhfr predicted drug failure. Only 3 instances were seen in the epidemic focus, but treatment failed in all. Only in the epidemic focus, 7 (88%) of 8 occurrences of > or = 1 mutations in dhps, and all occurrences of the Gly-437 allele of dhps, predicted failure. Association between mutations in dhps and mutations in dhfr was noted in the combined sites, irrespective of outcome. Although this makes the relationship of combined dhfr and dhps mutations to failure more difficult to interpret, it nevertheless supports S-P selection acting on both genes. In the holoendemic site, treatment success increased with age. In this location, acquired immunity may mask the impact of mutations in dhps, since sulfadoxine is a less effective treatment than pyrimethamine. PMID- 11491007 TI - Compliance to correct dose of chloroquine in uncomplicated malaria correlates with improvement in the condition of rural Nigerian children. AB - Non-compliance to correct dosing is thought to be one of the main causes of treatment failure of chloroquine in the home management of childhood malaria. There are few studies of compliance to drugs used for tropical diseases. In order to study compliance in the rural setting, chloroquine syrup was packaged with a novel pictorial insert for compliance to correct dosing. Compliance was assessed in a field trial in September 1996-December 1997, involving 632 children with uncomplicated malaria in Udi local government area in Nigeria. Written informed consent was obtained from mothers/guardians before children were enrolled in the study. There were 3 arms to the trial: control villages (group I) received chloroquine syrup without further intervention, group II received a pictorial insert with chloroquine syrup, and group III received chloroquine syrup, the pictorial insert and verbal instructions. Each group was made up of 3 health centres. Compliance was assessed by volumetric measurement of the chloroquine syrup left in 30-mL bottles and by questionnaires administered to mothers/helpers of the children. Control villages recorded full compliance for 36.5 +/- 4.4% of the children, group II for 51.9 +/- 7.9% and group III for 73.3 +/- 4.2%. There was a significant correlation (P < 0.0001) between full compliance, improvement and time for improvement of the condition. This study is deemed important because it focuses on children, who bear the greatest burden of malaria. It is unique for introducing a pictorial insert that illiterate villagers, who may not understand the use of age or weight in drug dispensing, may utilize as a substitute. PMID- 11491008 TI - Resistance of Plasmodium falciparum to antimalarial drugs in a highly endemic area of southern Viet Nam: a study in vivo and in vitro. AB - To assess the antimalarial sensitivity of Plasmodium falciparum in vivo and in vitro in a highly endemic area of southern Viet Nam, a field study was conducted (in 1999) at a rubber plantation in Binh Phuoc Province north of Ho Chi Minh City. Fifty patients were treated with either artesunate (4 mg/kg on day 0, then 2 mg/kg on day 1 to 4) or mefloquine (10 mg/kg at 0 h, then 5 mg/kg at 6 h), and their progress was followed for 28 days under standard WHO protocols. Blood spots were taken at baseline from all patients, as well as from those who redeveloped parasitaemia during follow-up, for polymerase chain reaction (PCR) determination of parasite genotypes to assist differentiation of re-infection from recrudescence. Both treatments cleared parasites within 5 days. Of the 25 mefloquine-treated patients, 2 (8%) re-presented with probable re-infections. For artesunate, 4 patients (16%) had re-infections and 5 (20%) had recrudescences. Sensitivity tests in vitro of pre-treatment P. falciparum isolates showed geometric mean IC50 values of 29, 38, 209 and 15 nmol/L for chloroquine (n = 32), mefloquine (n = 33), quinine (n = 31) and artemisinin (n = 31), respectively. There were significant correlations between IC50s for artemisinin and mefloquine (r = 0.72, P = 0.004), and chloroquine and quinine (r = 0.44, P = 0.05). These data show that, although mefloquine has been used for 10 years in Binh Phuoc Province, it remains fully effective, perhaps because an artemisinin derivative is commonly given at the same time. The recrudescence rate for artesunate is similar to those reported in other epidemiological contexts. The present in-vitro data imply that quinine remains effective and that reduced drug pressure has been associated with increased sensitivity of local strains of P. falciparum to chloroquine. Although from one hyperendemic area, these results may have implications for antimalarial prophylaxis and treatment strategies for residents and travellers to southern Viet Nam. PMID- 11491009 TI - Plasmodium falciparum: emerging mefloquine resistance in vitro in Para State, north Brazil. PMID- 11491010 TI - Long-term efficacy of single-dose combinations of albendazole, ivermectin and diethylcarbamazine for the treatment of bancroftian filariasis. AB - In a 'blinded' trial (in Sri Lanka, 1996-98) of 47 male asymptomatic microfilaraemic subjects with Wuchereria bancrofti infection, the safety, tolerability and filaricidal efficacy of 3 single-dose combination regimens were compared: albendazole 400 mg with ivermectin 200 micrograms/kg, albendazole 400 mg with diethylcarbamazine citrate (DEC) 6 mg/kg or albendazole 600 mg with ivermectin 400 micrograms/kg. Treated subjects were followed-up for 24 months. This represents the first long-term study using combinations of albendazole with DEC or ivermectin in the above doses against bancroftian filariasis. All subjects had pre-treatment microfilaria (mf) counts over 100/mL. All 3 treatments significantly reduced mf counts, with the albendazole-DEC-treated group showing the lowest mf levels at 18 and 24 months post-treatment. Filarial antigen tests suggested that all 3 treatments had significant activity against adult W. bancrofti; albendazole-DEC combination had the greatest activity according to this test, with antigen levels decreasing to 30.5% of pre-treatment antigen levels, 24 months after therapy. All 3 treatments were clinically safe and well tolerated. These results suggest that a single dose of albendazole 400 mg together with DEC 6 mg/kg is a safe and effective combination for suppression of microfilaraemia of bancroftian filariasis that could be considered for use in filariasis control programmes based on mass treatment of endemic populations. PMID- 11491011 TI - Placebo-controlled community trial of four cycles of single-dose diethylcarbamazine or ivermectin against Wuchereria bancrofti infection and transmission in India. AB - A double-blind placebo-controlled trial was carried out in 1994-98 to compare the effects of 4 cycles of single-dose diethylcarbamazine (DEC) or ivermectin on prevalence and geometric mean intensity (GMI) of microfilaraemia in the human population, infection rates in the vector population, and transmission intensity of Culex-transmitted Wuchereria bancrofti in rural areas in Tamil Nadu state, south India. Fifteen villages (population approximately 26,800) were included in the study: 5 villages each were randomly assigned to community-wide treatment with DEC or ivermectin or placebo. People over 14 kg bodyweight received DEC 6 mg/kg, ivermectin 400 micrograms/kg or a placebo, all identically packaged. After 2 cycles of treatment at a 6-month interval, the code was broken and the study continued as an open trial, with third and fourth cycles of treatment at a 12 month interval; 54-77% of eligible people (20,872) received treatment during the 4 cycles. Microfilaraemia prevalence and GMI fell by 48% and 65% with DEC and 60% and 80% with ivermectin respectively after 4 cycles of treatment. There was no change in the incidence of acute adenolymphangitis. Infection in resting mosquitoes fell significantly in all arms: 82%, 78% and 42% in the ivermectin, DEC and placebo arm, respectively. Landing mosquitoes also showed the same trend. The decline in infectivity was significant for resting (P < 0.05) and landing mosquitoes (P < 0.05) with ivermectin and DEC (P < 0.05), and for neither in the placebo group (P > 0.05). Transmission intensity was reduced by 68% with ivermectin and 63% with DEC. Transmission was apparently interrupted in 1 village with ivermectin, but infected resting mosquitoes were consistently found in this village. Single-dose community-level treatment with DEC or ivermectin is effective in reducing W. bancrofti infection in humans and mosquitoes, and may result in total interruption of transmission after several years of control. There is an immediate need to define the role of vector, parasite and community factors that influence the elimination of lymphatic filariasis, particularly the duration of treatment vis-a-vis efficacy of drugs, treatment compliance and efficiency of vectors. PMID- 11491012 TI - Booster effect of a new chromatographically purified Vero-cell rabies vaccine (CPRV): immunogenicity and safety of a single or double injection. PMID- 11491013 TI - Antiangiogenic and antitumor effects of a protein kinase C beta inhibitor in human hepatocellular and gastric cancer xenografts. AB - Hepatocellular carcinoma and gastric cancer are the most prevalent tumors worldwide. Hep3B hepatocellular carcinoma and HS746T gastric cancer were used as models for these diseases in culture and in vivo. The PKC beta inhibitor 317615.2HCl was not very cytotoxic toward HS746T or Hep3B cells in culture and was, in the main, additive in cytotoxicity with cisplatin, 5-fluorouracil and gemcitabine when cell in monolayer were exposed to these agents in combination with 317615.2HCl. Treatment of nude mice bearing HS746T or Hep3B xenografts with 317615.2HCl orally twice daily resulted in a small decreased in CD31-stainable intratumoral vessels in the HS746T tumors and 60% decrease in CD31-stainable vessels in the Hep3B tumors. Somewhat larger decreases were observed in the vessel stained with CD105. As a single agent 317615.2HCl produced tumor growth delays between 6.5 and 15 days in the HS746T xenograft and between 5 and 25 days in the Hep3B xenograft over the dosage range (3 to 30 mg/kg). Sequential and simultaneous combinations with 317615.2HCl and 5-fluorouracil and gemcitabine resulted in increases in tumor growth delay on both schedules. Gemcitabine produced a 15-day tumor growth delay of the HS746T gastric carcinoma that was increased to 40 days when combined simultaneously with 317615.2HCl and to 30 days with the sequential treatment regimen. 5-Fluorouracil produced a 9-day tumor growth delay of the Hep3B hepatocellular carcinoma that increased to 31 days by simultaneous treatment with 317615.2HCl and to 43 days with the sequential treatment regimen. Treatment with the protein kinase C beta inhibitor 317615.2HCl decreased HS746T and Hep3B angiogenesis and improved treatment outcome with 5 fluorouracil and gemcitabine. PMID- 11491014 TI - Dose and dose rate effects of whole-body gamma-irradiation: I. Lymphocytes and lymphoid organs. AB - The major goal of part I of this study was to compare varying doses and dose rates of whole-body gamma-radiation on lymphoid cells and organs. C57BL/6 mice (n = 75) were exposed to 0, 0.5, 1.5, and 3.0 Gy gamma-rays (60Co) at 1 cGy/min (low dose rate, LDR) and 80 cGy/min (high-dose rate, HDR) and euthanized 4 days later. A significant dose-dependent loss of spleen mass was observed with both LDR and HDR irradiation; for the thymus this was true only with HDR. Decreasing leukocyte and lymphocyte numbers occurred with increasing dose in blood and spleen at both dose rates. The numbers (not percentages) of CD3+ T lymphocytes decreased in the blood in a dose-dependent manner at both HDR and LDR. Splenic T cell counts decreased with dose only in HDR groups; percentages increased with dose at both dose rates. Dose-dependent decreases occurred in CD4+ T helper and CD8+ T cytotoxic cell counts at HDR and LDR. In the blood the percentages of CD4+ cells increased with increasing dose at both dose rates, whereas in the spleen the counts decreased only in the HDR groups. The percentages of the CD8+ population remained stable in both blood and spleen. CD19+ B cell counts and percentages in both compartments declined markedly with increasing HDR and LDR radiation. NK1.1+ natural killer cell numbers and proportions remained relatively stable. Overall, these data indicate that the observed changes were highly dependent on the dose, but not dose rate, and that cells in the spleen are more affected by dose rate than those in blood. The results also suggest that the response of lymphocytes in different body compartments may be variable. PMID- 11491016 TI - Presence of four stem cell populations in monolayer cultures derived from teratocarcinoma embryoid bodies. AB - In the present work, flow cytometry techniques together with morphologic studies were used to perform multiparametric analyses in cell cultures derived from CE44 teratocarcinoma embryoid bodies. The intrinsic cell parameters studied by flow cytometry were size (FALS), cytoplasmic complexity (ISS) and autofluorescence, expressed as LIGFL/FALS (green fluorescence intensity on a logarithmic scale/FALS). Our results showed that CE44 teratocarcinoma yields monolayers whose cells show a marked morphological heterogeneity and can be grouped according to flow cytometric criteria into four populations that remain stable throughout the entire time of culture. Moreover, these populations showed a different immunolabelling with the differentiation markers SSEA-1, TROMA-1 and anti vimentin. PMID- 11491015 TI - Dose and dose rate effects of whole-body gamma-irradiation: II. Hematological variables and cytokines. AB - The goal of part II of this study was to evaluate the effects of gamma-radiation on circulating blood cells, functional characteristics of splenocytes, and cytokine expression after whole-body irradiation at varying total doses and at low- and high-dose-rates (LDR, HDR). Young adult C57BL/6 mice (n = 75) were irradiated with either 1 cGy/min or 80 cGy/min photons from a 60Co source to cumulative doses of 0.5, 1.5, and 3.0 Gy. The animals were euthanized at 4 days post-exposure for in vitro assays. Significant dose- (but not dose-rate-) dependent decreases were observed in erythrocyte and blood leukocyte counts, hemoglobin, hematocrit, lipopolysaccharide (LPS)-induced 3H-thymidine incorporation, and interleukin-2 (IL-2) secretion by activated spleen cells when compared to sham-irradiated controls (p < 0.05). Basal proliferation of leukocytes in the blood and spleen increased significantly with increasing dose (p < 0.05). Significant dose rate effects were observed only in thrombocyte counts. Plasma levels of transforming growth factor-beta 1 (TGF-beta 1) and splenocyte secretion of tumor necrosis factor-alpha (TNF-alpha) were not affected by either the dose or dose rate of radiation. The data demonstrate that the responses of blood and spleen were largely dependent upon the total dose of radiation employed and that an 80-fold difference in the dose rate was not a significant factor in the great majority of measurements. PMID- 11491017 TI - Myxoid malignant fibrous histiocytoma of the uterus: a case with immunohistochemical, ultrastructural and tumor cell culture studies. AB - The clinical and pathological features, including ultrastructural and immunohistochemical findings, of a primary myxoid malignant fibrous histiocytoma of the uterus in a 60-year-old woman are reported. Microscopically, the principal feature of the tumor was a hypocellular area with diffuse degeneration, containing thin-walled curvilinear vessels, in which hyperchromatic small spindle and stellate cells, sometimes with vacuolated cytoplasm, were found. The transplanted tumor of primary cultured cells in nude mice presented as a prominent myxoid stroma confirming the histological structure of the primary tumor. Immunohistochemically, the presence of epithelial or heterogenous mesenchymal tumor components or cells of smooth muscle derivation were excluded and the tumor cells were positive for vimentin, CD 68, alpha 1-antitrypsin and alpha 1-antichymotrypsin. Ultrastracturally, pseudopodia and filopodia at the cell membrane and intracytoplasmic lysosomal granules were common. The patient had debulking surgery but died 38 days after the primary onset with the tumor occupying the entire abdomen and the pelvis. PMID- 11491018 TI - Vascular endothelial growth factor (VEGF) expression regulates angiogenesis accompanying tumor growth in a peritoneal disseminated tumor model. AB - Quantitative analysis of the process of tumor angiogenesis was performed in a new animal model of tumor microcirculation, in which colon carcinoma cells were inoculated into the peritoneal cavity of rats. Time-dependent changes in the microvascular architecture of mesenteric microvessels of tumor-bearing rats were visualized using an intravital microscope. Simultaneously, the expression of vascular endothelial growth factor (VEGF) by the tumor cells and VEGF secretion into ascites were analyzed. The results showed that VEGF increases microvascular permeability and stimulates the growth of microvessels into the tumor and that the spatial and temporal concentration of VEGF is strongly correlated. Such a correlation was stronger in the early angiogenic stages of tumor growth than in the subsequently occurring multiple metastatic stage, when VEGF was still observed at a high level in tumor surroundings. Thus, VEGF is suggested to be primarily involved in the pathophysiological control of angiogenesis accompanying tumor progression. PMID- 11491019 TI - Reconstituted normal human breast in nude mice: estrogen and progesterone receptors regulation. AB - Actions of estrogen and progesterone on normal human breast are mediated by their respective receptors. Since receptor regulation studies in humans are difficult to perform, we have utilized our model system of reconstituted normal human breast in nude mice to determine expression of estrogen and progesterone receptors in response to exogenous estrogen and progesterone. In normal human breast in situ, only a subset of epithelial cells (about 10%) are positive for estrogen receptor. These cells which remain positive after transplantation into nude mice can be completely downregulated by administration of exogenous estrogen resulting in all transplanted human breast epithelial cells now being negative for estrogen receptor. Exogenous estrogen also upregulates the progesterone receptor in a subset of human breast epithelial cells rather than in all cells. Exogenous progesterone, alone or in combination with estrogen, did not affect the estrogen or progesterone receptors. Double immunofluorescent labelings for a proliferation marker and progesterone receptor in both surgical specimens and nude mice transplants demonstrate that proliferation and progesterone receptor expression do not take place in the same cell. Our in vivo model system enables studies on the biology of primary normal human breast epithelial cells which otherwise are difficult to perform in humans. PMID- 11491020 TI - A modified method for the detection of differentially expressed mRNAs without using radioactivity. AB - We present here a modification of the original differential display approach using a single oligo(dT) primer for the reverse transcription reaction (instead of the various oligo(dT)NM primers that subdivide the pool of mRNAs) and a combination of 25-mer or 26-mer arbitrary primers together with 30-mer anchored primers for the PCR reaction. The PCR products are, then, efficiently separated in a non-denaturing polyacrylamide gel and the bands are visualized after staining with silver nitrate. The model for the development of our differential display approach was seven clones of an insect species: the aphid Myzus pesicae (Sulzer) (Homoptera: Aphididae). We believe that our modified differential display technique, with the efficient resolution of the DNA bands in a non denaturing gel and staining with silver can be applied as an alternative non radioactive detection of differentially expressed messages in various cell populations. In addition, the method could be used as a supplementary tool to other techniques for examining inter- and intraspecific genetic variation in aphids. PMID- 11491021 TI - Diverse biological activities of moxa extract and smoke. AB - Diverse biological activities of moxa extracts and smoke (gas phase) were investigated. Moxa was extracted with hot water (Fr. I), or ethanol (Fr. II), or extracted with hot water after ethanol wash (Fr. III) and then lyophilized to obtain the dried powders. Moxa smoke (containing a lot of gaseous components obtained by burning Moxa) (Fr. IV) was collected into phosphate-buffered saline and quantified spectrophotometrically. These extracts and Moxa smoke showed comparable cytotoxic activity against human oral tumor cell lines (HSC-2, HSG). Human gingival fibroblasts (HGF) were more resistant to any Moxa fractions. Neither of the extracts showed anti-HIV activity. Pretreatment of mice with Fr. I significantly reduced the lethal effect of E. coli infection. All extracts produced radicals under alkaline condition, with a maximum intensity at pH 10.5, and enhanced the radical intensity of sodium ascorbate. It was unexpected that these extracts show significant O2- scavenging activities. These data suggest the medicinal efficacy of Moxa extracts and smoke. PMID- 11491022 TI - Clinician perseverance. Helping patients overcoming tobacco dependence. PMID- 11491023 TI - Firstbreath. A collaborative effort to help Wisconsin's pregnant women stop smoking. PMID- 11491024 TI - Where there's smoke, there's help. PMID- 11491025 TI - Physicians key in reaching teen smokers. PMID- 11491026 TI - Trends in self-reported cigarette smoking, Wisconsin, 1984-1999. AB - The purpose of this study is to assess trends in self-reported cigarette smoking among adults in Wisconsin. Behavioral Risk Factor Survey data from 1984-1999 were analyzed to determine changes in smoking prevalence. Results of this analysis show almost no change in overall adult smoking prevalence over the past 15 years. Smoking prevalence rates did, however, decrease slightly for men from 29% in 1984 to 22% in 1999. Moreover, smoking prevalence for adults 18 to 24 years of age increased from 25% in 1994 to 36% in 1999. These results show that despite state and national efforts over the past 15 years to reduce cigarette smoking among adults, minimal progress has been made. Furthermore, the increase in smoking rates among 18 to 24 year olds highlights the need for additional prevention and cessation efforts targeted toward this group. Understanding past trends in cigarette smoking is critical for the Wisconsin Tobacco Control Board and other public health advocates to monitor progress toward the goal of reducing tobacco use. PMID- 11491027 TI - Cigarette smoking in Wisconsin: the influence of race, ethnicity, and socioeconomics. AB - A disparate burden of cigarette use has been demonstrated among demographic subgroups both in the United States and Wisconsin. We examined patterns of adult current smoking prevalence in Wisconsin by race, Hispanic ethnicity, household income, and education to assess whether differences exist among these subgroups. This analysis revealed a strong graded relationship between household income, education, and smoking prevalence, consistent among non-Hispanic whites and blacks, though not Hispanics. Respondents with less than a high school education had significantly higher smoking prevalence rates (41%) than those with a college degree or more (13%). Smoking prevalence rates did not significantly differ between the race and ethnicity subgroups overall, or by gender and education, although they differed in some age and income subgroups. Possible explanations for the socioeconomic gradient include differences in tobacco product marketing practices, indoor workplace smoking policies, and access to health information, resources, and consistent, high-quality health care. PMID- 11491028 TI - Smoking during pregnancy in Wisconsin compared to the United States, 1997. AB - PURPOSE: The purpose of this study is to identify disparities in maternal smoking between Wisconsin and United States women and to determine differences that explain the higher percentage of pregnant women who smoke in Wisconsin compared to the United States. METHODS: 1997 Wisconsin and US birth certificate data were compared and stratified by age, education, and race/ethnicity. The relative risks (the risk of Wisconsin women smoking during pregnancy compared to US women smoking during pregnancy) were calculated by direct standardization to the 1997 US distribution for these characteristics. RESULTS: In 1997, 17.9% of Wisconsin women smoked during pregnancy, compared with 13.2% nationally (relative risk [RR] = 1.4; Wisconsin women were 40% more likely to smoke than US women). Age and education adjusted RRs among Wisconsin women aged 20 and older were 2.3 (American Indian), 2.8 (Hispanic), and 2.2 (non-Hispanic black), while the RR was 1.2 for non-Hispanic white mothers. Among women 20 and older, the crude RR for Wisconsin was 1.5; adjusting for age, education, and race/ethnicity only slightly decreased the RR to 1.4. CONCLUSIONS: The percentage of women who smoke during pregnancy in Wisconsin continues to be above the national rate and the Healthy People 2000 goal of 10% or less. Disparities with the US average are particularly great for minority women in Wisconsin. Adjusting for age, education and ethnicity does not explain Wisconsin's higher prenatal smoking rate. PMID- 11491029 TI - Research shows effectiveness of safety belts. PMID- 11491030 TI - Physicians need to listen to patients. PMID- 11491031 TI - Tobacco use among American Indian/Alaska Native youth in Wisconsin. AB - The Great Lakes Inter-Tribal Council investigated the level of tobacco use among American Indian/Alaska Native (AI/AN) youth to help Tribal communities appropriately target tobacco prevention and intervention efforts. A youth tobacco survey of AI/AN middle school students in Wisconsin was conducted. Nine hundred six AI/AN students in grades 6-8 at 15 tribal and public schools were surveyed with the Great Lakes Inter-Tribal Council Youth Tobacco Survey (GLITC YTS) Overall, 79% of students reported having tried some tobacco product; 45% reported currently using some form of tobacco. Tobacco use among AI/AN middle school students was higher than that of middle school students participating in the Wisconsin Youth Tobacco Survey. PMID- 11491032 TI - Tobacco use trends and correlates among students in the Madison Metropolitan School District. AB - Trends in cigarette use among students in grades 7-12 in the Madison Metropolitan School District were examined using data from the Dane County Youth Survey, which has been conducted every 5 years since 1980. Past year use of smoking tobacco among these students declined from 40% to 45% through 1995 to 32% in 2000. Past month cigarette use declined from 27% in 1995 to 18% in 2000; daily smoking dropped from 15% in 1990 to 8% in 2000. Among 12th grade students, 1999 national data show that 35% smoked in the past month, compared to 30% of Madison 12th graders. Bivariate analyses show significant socio-economic and race/ethnic disparities in cigarette smoking rates. Hispanic, Native American and white students reported higher rates of smoking than Asian and African American students. Students whose parents had more education were less likely to smoke, as were those students who themselves were academically successful and who had college plans. Students who smoked were far more likely than non-smokers to engage in other risky behaviors, including use of alcohol and marijuana, and to be sexually active. Parental monitoring and supervision, support in one's neighborhood, and participation in volunteer and leadership activities were associated with lower rates of smoking. Risk factors associated with increased rates of smoking included other drug use, aggressive behavior, less family disapproval of risky behavior, and self-concerns about one's own behaviors. These data can assist in targeting prevention and cessation efforts for Wisconsin adolescents. Physicians and other care providers should consider cigarette smoking as a sentinel event that signals the need for screening for potential substance abuse, sexually transmitted diseases (STDs) and contraceptive care. School-based preventive efforts should be continued, particularly at the middle school level, with cessation programs added at the high school level. PMID- 11491033 TI - Progress in reducing per capita cigarette sales in Wisconsin, 1985-1999. AB - Over the past several decades, initiatives have been undertaken both on a national and state level to reduce cigarette smoking and its resultant health consequences. We compared trends in per capita cigarette sales in Wisconsin versus other states from 1985-1999. During this time period, per capita cigarette sales declined 21% in Wisconsin, versus 26% in the United States. Although the average per capita sales in Wisconsin were less than sales in the United States in both 1985 (107 versus 122 packs per capita) and 1999 (84 versus 90 packs per capita), Wisconsin's sales have declined at a slower rate, narrowing the gap between Wisconsin and US sales. Other states, such as California and Massachusetts, with large statewide tobacco control programs had rates of decline in cigarette sales over twice the Wisconsin decline from 1985-1999. In 1985, only 10 states had lower per capita sales than Wisconsin. By 1999, 24 states had lower rates, indicating greater relative progress in reducing sales in other states. Possible explanations for the greater decline in per capita cigarette sales in other states include differences in tobacco control programs, tobacco excise tax increases, and other tobacco policy initiatives. PMID- 11491034 TI - Medipay for medical bills. PMID- 11491035 TI - Under-use of tobacco dependence treatment among Wisconsin's fee-for-service Medicaid recipients. AB - BACKGROUND: Wisconsin Medicaid enrollees are eligible for treatment for tobacco dependence at minimal charge to the enrollee. This paper describes an evaluation of the use of this treatment within the Wisconsin fee-for-service Medicaid program. METHODS: Pharmaceutical claims data for Medicaid fee-for-service patients were analyzed for the year 1999 to determine rates of treatment use. RESULTS: Of 261,435 adults enrolled in fee-for-service Medicaid for 1 or more months, only 1131 adults received pharmacotherapy for tobacco dependence in 1999 at a modest cost of approximately $135 per treatment user. This represents less than 2% of the adult Medicaid fee-for-service patients who smoke. DISCUSSION: Few Medicaid fee-for-service enrollees are receiving evidence-based treatment for tobacco dependence, the leading preventable cause of illness and death in Wisconsin. The 16,000 Wisconsin physicians caring for these patients are urged to intervene with every Medicaid patient who smokes. To assist in this effort, the Wisconsin Medicaid program's coverage for tobacco dependence treatment is explained and a series of myths are corrected. PMID- 11491036 TI - Meeting the challenge of tobacco use within the Medicaid population. AB - The Wisconsin Medicaid Program, which purchases health care for Wisconsin's low income residents, has entered into a number of partnerships designed to systematically address tobacco use by Medicaid recipients. With the help of Wisconsin HMOs, a chart audit established an overall tobacco use prevalence of 45.7%, about double that of the general population. A direct survey of Medicaid recipients confirmed a rate of 45.0%. The chart audit also revealed a dramatic under-utilization of currently available Medicaid smoking cessation benefits. Plans are now being made to increase this utilization on a statewide basis. In addition, a partnership with the Wisconsin Women's Health Foundation and the Bureau of Family and Community Health Program within the Division of Public Health has resulted in a pilot project to address tobacco use by low income, pregnant women. Finally, a partnership with a specialized HMO serving those with disabilities and the Mental Health Association in Milwaukee County plans to address tobacco use by those with chronic and persistent mental illness. PMID- 11491037 TI - Smoking policies in manufacturing and assembly workplaces, Wisconsin, 1999. AB - Although Wisconsin's clean indoor air law prohibits or restricts smoking in certain areas, it specifically exempts manufacturing and assembly workplaces from its provisions. We conducted a mail survey of 1500 randomly selected employers to determine the nature and extent of smoking policies in Wisconsin's blue-collar workforce. Of the 1042 (70%) respondents, 49% prohibit all smoking; 26% allow smoking only in designated areas; 18% allow smoking in all areas except designated non-smoking areas; and 7% allow smoking anywhere. Larger employers were more likely to have smoking policies. Of the 61% of respondents who indicated having a formal smoking policy, the reasons for having the policy were safety (40%), health (38%), or employee request (12%). For those employers without a smoking policy, the main reasons were that few employees smoke (37%), the decision is left to the employee's discretion (32%), or employees may object to having a policy (10%). About half of the employees in manufacturing and assembly workplaces continue to be exposed to environmental tobacco smoke. PMID- 11491038 TI - Lung cancer mortality report card: measuring progress in Wisconsin's counties, 1979-1998. AB - PURPOSE: To compare and contrast trends in lung cancer mortality in Wisconsin's 72 counties, as a measure of long-term progress in tobacco control. METHODS: Lung cancer mortality data were abstracted from the Centers for Disease Control and Prevention's WONDER database from 1979 to 1998. Percent change in lung cancer mortality rates were analyzed at the county and national levels from 1979-1983 to 1994-1998. RESULTS: In Wisconsin, lung cancer mortality rates increased 23%, compared to a 19% increase in the United States. There was more variation in lung cancer mortality trends between Wisconsin counties than between states, with increases in 67 of Wisconsin's 72 counties. Lung cancer mortality rates increased for men in 44 counties and for women in 68 counties. Only 4 counties in Wisconsin had fewer lung cancer deaths in 1994-1998 compared with 1979-1983. CONCLUSIONS: Progress in reducing lung cancer mortality in Wisconsin--a long-term measure of progress in tobacco control--has lagged behind the rest of the United States. Nevertheless, some Wisconsin counties have experienced more progress in reducing the health burden from lung cancer, suggesting that differences exist between communities in the effectiveness of their tobacco control efforts. PMID- 11491039 TI - Solving complex health problems with interdisciplinary approaches. PMID- 11491040 TI - Ask, assess, advise, assist, arrange are keys to smoking cessation. PMID- 11491041 TI - Men's health matters: a gender in crisis. PMID- 11491042 TI - Ingested foreign bodies in the paediatric patient. AB - BACKGROUND: Paediatric foreign body (FB) ingestion is a common problem and while most can be managed conservatively, a sub-population require intervention. AIMS: To establish clear guidelines for management of paediatric FB ingestion. METHODS: A retrospective chart review analysing all paediatric admissions with FB ingestion over a 10-year period from 1990 to 1999. RESULTS: Of 339 patients presenting to the accident and emergency department with FB ingestion, 59 required admission. Ingestion was accidental in 93.0% of patients. The reasons for admission were as follows: large FBs; dangerous FBs; and living far from the hospital. Nineteen patients (32.2%) were discharged without intervention. Thirty seven (62.7%) required endoscopic retrieval. In two, the FB was not identified at endoscopy. Only three (5%) required surgery. CONCLUSION: Conservative management of FB ingestion in the paediatric population is possible in the majority of cases. However, a minority require intervention. While guidelines for intervention are ill-defined, definitive indications include symptomatic patients, or dangerous objects. PMID- 11491043 TI - Outcome of infants born to hepatitis C infected women. AB - BACKGROUND: Hepatitis C virus (HCV) can be transmitted vertically from mother to infant, either late in pregnancy or at delivery. AIMS: To determine the outcome of infants born to HCV infected women, to characterise epidemiology and to design an appropriate infant monitoring schedule. METHODS: Three hundred and fourteen infants, born to 296 HCV positive women between 1994 and 1999 were monitored for a median of 18 months (range 1-52). RESULTS: Forty per cent of infants were small for age and 46% had neonatal abstinence syndrome (NAS). Of 173 infants of defined status, 11 were infected (vertical transmission rate [VTR] 6.4%, 95% CI 2.8-10). Infected infants were diagnosed at a median of three months (range 0.5-10). Liver transaminases elevation was documented in 8% of uninfected infants. A negative HCV PCR test before one month of age did not exclude infection but all infected patients had detectable HCV RNA when next tested (range 2-10 months). CONCLUSIONS: 94% of infants born to HCV antibody positive women are not HIV infected. Liver transaminase elevation in exposed infants is not always indicative of infection. A minimum monitoring schedule of testing (PCR and antibody) at six to eight weeks, six and 18 months allows early diagnosis while detecting late seroconversions. PMID- 11491044 TI - Soluble adhesion molecules in inflammatory bowel disease. AB - BACKGROUND: Soluble adhesion molecules are elevated in a number of inflammatory conditions. AIMS: To investigate the correlation of soluble intercellular adhesion molecule-1 (sICAM-1) and sE-selectin with the activity of inflammatory bowel disease (IBD). METHODS: sICAM-1 and sE-selectin were measured by an enzyme linked immunosorbent assay (ELISA) in 53 patients with ulcerative colitis (UC) and 38 patients with Crohn's disease (CD). RESULTS: Patients with active UC and CD had significantly higher sICAM-1 than patients with inactive disease and controls. Patients with pancolitis had significantly higher levels than patients with distal colitis. There was a significant difference in sE-selectin levels between patients with active CD and control sICAM-1. sE-selectin did not correlate with the Harvey Bradshaw index (HBI). C-reactive protein (CRP) and microalbuminuria were better markers than sICAM-1 or sE-selectin which correlated with serum tumour necrosis factor (TNF)-alpha. CONCLUSION: sICAM-1 and sE selectin are elevated in the serum of patients with IBD but CRP and microalbuminuria reflect clinical disease activity more accurately. This study does not support the routine use of soluble adhesion molecules as disease activity markers in IBD. PMID- 11491045 TI - Inhibitory effect of IL-4 on the production of IL-1 beta and TNF-alpha by gastric mononuclear cells of Helicobacter pylori infected patients. AB - BACKGROUND: The mechanisms by which Helicobacter pylori damages host tissues are complex and unclear. AIMS: To determine the effect interleukin (IL)-4 and interferon-gamma (IFN-gamma) on the production of proinflammatory cytokines by the gastric mononuclear cells of H. pylori infected patients was determined. METHODS: The effect of IL-4 and IFN-gamma on the production of proinflammatory cytokines by the gastric mononuclear cells of H. pylori infected patients was determined. RESULTS: IL-4 markedly reduced the production of IL-1 beta and tumour necrosis factor-a (TNF-alpha) by the gastric mononuclear cells of H. pylori infected patients (P < 0.01). Enzyme-linked immunospot (ELISPOT) assay indicated a decrease in IL-4 producing cells (P < 0.05) and an increase in IFN-gamma secreting cells (P < 0.01). CONCLUSIONS: The increased level of proinflammatory cytokines may be due to hyposecretion of IL-4 in H. pylori infected patients. T helper type 1 (Th1) immune response with increased IFN-gamma also contributes to the inflammation of the gastric mucosa. PMID- 11491047 TI - Scabies management in the community. AB - BACKGROUND: Unidentified and undertreated cases of scabies are the source of continued spread. AIMS: To evaluate management of scabies in the community. METHODS: Eighty-six outpatient and ward referrals were reviewed. Fifty-six had previously applied scabicidal treatments (Group 1), whilst in 30 patients the diagnosis was made de novo (Group 2). Treatment failure was identified by a detailed questionnaire. In both groups, patients were reviewed at four to six weeks. RESULTS: Sixty-four per cent of Group 1 was initially correctly diagnosed. Seventy per cent of Group 1 patients had received verbal and written instruction as part of previous unsuccessful treatments. The clearance rate in the combined group treated according to protocol was 96%; 100% clearance was ultimately achieved. CONCLUSION: Inconsistency in diagnosis and treatment of patients is common. A treatment protocol should be made available to family practitioners. PMID- 11491046 TI - Premature hair greying may predict reduced bone mineral density in Graves' disease. AB - BACKGROUND: Premature hair greying has been associated with low bone mineral density (BMD), and it may be more frequent in Graves' disease. AIMS: To determine whether premature greying is associated with reduced BMD in women with Graves' disease and in control women, and to examine whether premature greying is more common in Graves' disease. METHODS: Premature greying (> 50% grey by 40 years) and BMD were determined in 44 women with a history of Graves' disease and 133 female controls referred for routine BMD measurement. Exclusion criteria included diseases or drugs known to affect BMD. RESULTS: Mean Z and T scores at the lumbar spine were significantly lower (P < 0.04) in subjects with premature greying than in those not prematurely grey among women with Graves' disease, but not among control women. Multiple regression confirmed this difference between Graves' and control women (P = 0.041). There were no differences at other measurement sites. Of Graves' patients, 36% were prematurely grey compared with 25% of control women (P = 0.14). CONCLUSION: Premature greying may be a weak marker for reduced BMD in women with a history of Graves' disease, but it is not a marker in normal women. PMID- 11491048 TI - Designing a medical Web site. AB - BACKGROUND: A web site is a valuable shop window for any medical unit with something to sell or something to say. AIMS: The aim of this report is to outline the basic steps of web page design for the individual or unit with limited financial resources. METHODS: There are two ways of designing a web site. A reputable web design company can be employed, but this is usually expensive. Alternatively, a web site can be designed in-house using commercial software, following a few simple steps. The basic requirements are a personal computer, software that is available on most computers and access to a few peripheral items of hardware. An outline of the page design should first be put down on paper. This can be transferred to a computer file using a web page design program. This file is then sent to a server for publication on the World Wide Web (WWW). CONCLUSION: Designing and publishing a web page can take time and effort, but the rewards can be great and the results will reflect the message and motto of the unit. PMID- 11491051 TI - John Mallet Purser (1839-1929). PMID- 11491049 TI - Electrogastrography: a non-invasive measurement of gastric function. AB - BACKGROUND: Electrogastrography (EGG) is the non-invasive measurement of gastric electrical activity. With the development of modern technology, improved recording and automated analysis, it is a reliable and accurate technique for the measurement of gastric myoelectrical activity providing information about the frequency and regularity of the gastric slow wave. AIM: The aim of this report is to evaluate its role in clinical practice. METHODS: The literature is reviewed and its role investigated. RESULTS: EGG has been successfully used in the investigation of gastroparesis, non-ulcer dyspepsia (NUD), gastric emptying (GE) disorders and diabetes mellitus (DM). EGG also provides an insight into the effect of medications on gastric function, e.g. edrophonium, cisapride, erythromycin and proton-pump inhibitors (PPI). CONCLUSIONS: EGG has a developing role in the assessment of gastric dysfunction and on the effect of medical treatment. The effect of surgery and anaesthesia on gastric myoelectric activity is less clear. PMID- 11491052 TI - The mythology of medicine. PMID- 11491053 TI - Meckel's diverticulum present as pyogenic liver abscess: report of first case. PMID- 11491050 TI - The role of apoptotic cell death in cardiovascular disease. AB - BACKGROUND: Programmed cell death, or apoptosis, is a distinct, managed form of cell death. It is fundamentally different from necrosis. It is a genetically controlled, energy-dependent method of cellular deletion without inflammation. In the cardiovascular system, apoptosis occurs as a primary and secondary event in disease pathogenesis. This review addresses our current understanding of the initiation, propagation and significance of apoptosis in the cardiovascular system, as well as assessing therapeutic potentials arising therefrom. METHODS: A Medline search was performed and relevant publications reviewed. Further articles were obtained from the references of these publications. RESULTS: and conclusions Apoptotic cell death is a key element in the pathogenesis and progression of ischaemia-reperfusion (IR) injury, cardiac failure, myocardial infarction, atherosclerosis, endothelial dysfunction and the clinical syndromes which these situations produce. Our increased understanding of the role of apoptosis in the pathogenesis of cardiovascular disease offers potential to develop new therapeutic strategies. PMID- 11491054 TI - Universal precautions, preventing infection and protecting staff. PMID- 11491055 TI - Meralgia paresthetica following total hip replacement. PMID- 11491056 TI - Stethoscope versus ophthalmoscope? PMID- 11491057 TI - Host cytokine responses to Helicobacter pylori: an important determinant of clinical outcome. PMID- 11491058 TI - Scabies management in the community. PMID- 11491061 TI - Neuroprotection of the infant brain--how many roads? Proceedings of a workshop. London, United Kingdom, 9-10 February 2000. PMID- 11491060 TI - Day case laparoscopic cholecystectomy is feasible. AB - BACKGROUND: Laparoscopic cholecystectomy is the operation of choice for cholelithiasis. AIMS: The aims of our study were to assess the feasibility of day case laparoscopic cholecystectomy (DCLC) in selected patients. METHODS: DCLC was introduced in this unit in July 1999. The first 50 patients were prospectively evaluated up to February 2001. RESULTS: All patients were under 55 years of age with an ASA grade of I (n = 48) or II (n = 2). The mean age was 41.1 years (range 20-55 years) and the male:female ratio was 1:6. All patients had a standard anaesthetic protocol. Patients were discharged 10 to 12 hours postoperatively with a pro forma, which was reviewed at one week in the clinic. The conversion rate was 2%. Three required overnight admission due to excessive nausea, hypertension and for an unforeseen psychosocial problem. Ninety per cent of patients were suitable for same day discharge. No patient required subsequent readmission. CONCLUSION: DCLC is feasible and safe in carefully selected patients and has the advantages of convenience and cost-effectiveness. PMID- 11491059 TI - Deliberate self-poisoning in an Irish county hospital. AB - BACKGROUND: In 1997, 433 people committed suicide in Ireland, one-quarter of whom were less than 24 years of age. AIM: To determine demographics, agent choice and source, suicidality and follow-up care of deliberate self-poisoning patients. METHOD: Details of 111 patients admitted to one hospital in 1997 following drug overdose were studied retrospectively. Eleven had been accidental ingestions, the remaining 100 were deliberate self-poisoning. RESULTS: Men accounted for 38% of the presentations and were more likely to have suicidal intent than women. An average of 2.3 different agents were used. Paracetamol was taken by 37%, hypnotics/anxiolytics by 33% and nonsteroidal anti-inflammatories by 17%. Alcohol was consumed synchronously by 51% and 17% fulfilled criteria for alcohol dependency. One-third of patients were clinically depressed. All six patients requiring ventilation had consumed a combination of tricyclic antidepressants and alcohol. There were no deaths. CONCLUSION: Deliberate self-poisoning remains a significant problem. Paracetamol and alcohol use are particularly marked in this population. The combination of tricyclic antidepressant drugs and alcohol were the most dangerous. PMID- 11491062 TI - [Current problems in animal welfare. Hannover, Germany, October 12-13, 2000. Proceedings]. PMID- 11491063 TI - ["Diagnosis related groups" in natural medicine]. PMID- 11491064 TI - Proceedings of the workshop on epilepsy and learning disabilities. Troina, Italy, April 28-30, 2000. PMID- 11491065 TI - Immunocompromised Host Society Consensus Conference on Epidemiology, Prevention, Diagnosis, and Management of Infections in Solid-Organ Transplant Patients. Davos, Switzerland, 23 June 1998, fully updated summer 2000. PMID- 11491066 TI - A commitment to safety. Tools for implementing JCAHO's new patient safety standards. PMID- 11491067 TI - Meeting challenges of professional certification. PMID- 11491068 TI - HCFA revises proposed ABN (advanced beneficiary notice) for labs. PMID- 11491069 TI - Proceedings of the 11th International Congress of Immunology. 22-27 July 2001, Stockholm, Sweden. PMID- 11491070 TI - The role of teaching the doctor-patient relationship in medical education. PMID- 11491071 TI - Current readings in nuclear medicine. PMID- 11491072 TI - Curing potential of dual-polymerizable resin cements in simulated clinical situations. AB - STATEMENT OF PROBLEM: Little is known about the ability of dual-polymerizable resin cements to polymerize when they are used in various clinical scenarios. Purpose. This study was conducted to determine whether any of 6 commercially available dual-polymerizable resin cements should be classified as an "all purpose" resin cement. MATERIAL AND METHODS: Chemical conversion values (C=C converted to C-C, or the extent of the curing reaction) of 6 commercially available dual-polymerizable resin cements were determined with infrared spectroscopy in 5 clinically relevant scenarios. Scenarios included: using each cement in a dual-polymerizable mode (mixing of 2 pastes); light polymerizing curing through Mylar sheets (dual-Mylar), which served as the control; light polymerizing through 3-mm porcelain (dual-3 mm); and no exposure to light (dual no light). The single-component light-polymerizable product was also tested as follows: exposed directly through Mylar (light-Mylar) or exposed through 3 mm of porcelain (light-3 mm). RESULTS: For each product, dual-Mylar treatment yielded the highest conversion value of all treatments (control for each product). For all products, dual-3 mm conversion was at least 97% of control and equivalent to control, with the exception of Lute-It!. Dual-no light conversion was less than control treatment but at least 86% of control for all products except for Variolink II (62% of control). For all products in dual-no light mode, except Choice and Variolink II, conversion was at least equal to the light-Mylar values. Only 1 product (Variolink II) did not demonstrate increased conversion values for dual-Mylar compared with light-Mylar treatments. For most other products (Calibra, Insure, and Lute-It!), conversion values for light-3 mm were significantly less than for light-Mylar. Conversion values for Nexus, Choice, and Variolink II were equivalent between light-Mylar and light-3 mm treatments. CONCLUSION: The choice of a dual-polymerizable cement should be based on its intended use because not all products polymerize adequately in every clinical situation. Although no cement met the stated criteria for an "all-purpose" cement, those tested did produce a range of product-specific results. PMID- 11491073 TI - Alternative uses of a visible light-polymerized material. AB - Three different procedures that involve the use of Triad gel are described. This light-polymerized resin material is versatile and easy to use in the dental clinic and laboratory. The procedures described include the transfer of ridge topography for ovate pontic sites, implant lab analog fixation, and solid/soft tissue cast fabrication. PMID- 11491074 TI - Quality issues in otorhinaryngology: Part I. PMID- 11491075 TI - Non-invasive mechanical ventilation. PMID- 11491076 TI - BTS asthma guide. PMID- 11491077 TI - Asthma and early childhood infectious disease. Critical time for protective effect of large family on asthma may not be during first year of life. PMID- 11491078 TI - Winter deaths: warm housing is not enough. PMID- 11491079 TI - A Desulfitobacterium strain isolated from human feces that does not dechlorinate chloroethenes or chlorophenols. AB - An anaerobic bacterium, strain DP7, was isolated from human feces in mineral medium with formate and 0.02% yeast extract as energy and carbon source. This rod shaped motile bacterium used pyruvate, lactate, formate, hydrogen, butyrate, and ethanol as electron donor for sulfite reduction. Other electron acceptors such as thiosulfate, nitrate and fumarate stimulated growth in the presence of 0.02% yeast extract and formate. Acetate was the only product during fermentative growth on pyruvate. Six mol of pyruvate were fermented to 7 mol of acetate. 13C NMR labeling experiments showed homoacetogenic 13C-CO2 incorporation into acetate. The pH and temperature optimum of fermentative growth on pyruvate was 7.4 and 37 degrees C, respectively. The growth rate under these conditions was approximately 0.10 h(-1). Strain DP7 was identified as a new strain of Desulfitobacterium frappieri on the basis of 16S rRNA sequence analysis (99% similarity) and DNA-DNA hybridization (reassociation value of 83%) with Desulfitobacterium frappieri TCE1. In contrast to described Desulfitobacterium strains, the newly isolated strain has not been isolated from a polluted environment and did not use chloroethenes or chlorophenols as electron acceptor. PMID- 11491080 TI - Further studies on RpoS in enterobacteria: identification of rpoS in Enterobacter cloacae and Kluyvera cryocrescens. AB - RpoS, the alternative sigma factor sigma(s), is important for bacterial survival under extreme conditions. Many enterobacteria are opportunistic human pathogens and their ability to survive in a changing environment could be an essential step for their virulence. To determine the presence of this gene in enteric bacteria, an Escherichia coli rpoS probe was constructed and used to detect the presence of this gene in different species. A gene homologous to rpoS was found in Citrobacter amalonaticus, Enterobacter cloacae, Klebsiella planticola, Kluyvera cryocrescens, Serratia rubidaea, Shigella sonnei, and Yersinia ruckeri. Providencia stuartii and Proteus vulgaris were the only tested enterobacteria that did not show any signal with the E. coli rpoS probe or that did not lead to amplification of an rpoS fragment using specific primers. The rpoS gene from E. cloacae and from K. cryocrescens was cloned and sequenced and a mutant allele was constructed in E. cloacae. Survival rates under different harsh conditions were followed in order to determine the effect of rpoS inactivation in exponential- and stationary-phase cells of both strains. E. cloacae rpoS mutants were more sensitive to extreme pH, high osmolarity, and high temperature than the wild type. PMID- 11491081 TI - Isolation and characterization of a thermostable intracellular enzyme with peroxidase activity from Bacillus sphaericus. AB - During a screening for bacteria producing enzymes with peroxidase activity, a Bacillus sphaericus strain was isolated. This strain was found to contain an intracellular enzyme with peroxidase activity. The native enzyme had a molecular mass of above 300 kDa and precipitated at a salt concentration higher than 0.1 M. Proteolytic digestion with trypsin reduced the molecular mass of the active enzyme to 13 kDa (dimer) or 26 kDa (tetramer) and increased its solubility, allowing purification to homogeneity. Spectroscopic investigations showed the enzyme to be a hemoenzyme containing heme c as the covalently bound prosthetic group. The enzyme was stable up to 90 degrees C and at alkaline conditions up to pH 11, with a pH optimum at pH 8.5. It could be visualized by activity staining after SDS-PAGE and showed activity with a number of typical substrates for peroxidases, e.g., 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) diammonium salt, guaiacol and 2,4-dichlorophenol; however the enzyme had no catalase and cytochrome c peroxidase activity. PMID- 11491082 TI - Cell compartmentalisation in planctomycetes: novel types of structural organisation for the bacterial cell. AB - The organisation of cells of the planctomycete species Pirellula marina, Isosphaera pallida, Gemmata obscuriglobus, Planctomyces maris and "Candidatus Brocadia anammoxidans" was investigated based on ultrastructure derived from thin sections of cryosubstituted cells, freeze-fracture replicas, and in the case of Gemmata obscuriglobus and Pirellula marina, computer-aided 3-D reconstructions from serial sections of cryosubstituted cells. All planctomycete cells display a peripheral ribosome-free region, termed here the paryphoplasm, surrounding the perimeter of the cell, and an interior region including any nucleoid regions as well as ribosome-like particles, bounded by a single intracytoplasmic membrane (ICM), and termed the pirellulosome in Pirellula species. Immunogold labelling and RNase-gold cytochemistry indicates that in planctomycetes all the cell DNA is contained wholly within the interior region bounded by the ICM, and the paryphoplasm contains no DNA but at least some of the cell's RNA. The ICM in Isosphaera pallida and Planctomyces maris is invaginated such that the paryphoplasm forms a major portion of the cell interior in sections, but in other planctomycetes it remains as a peripheral zone. In the anaerobic ammonium oxidising ("anammox" process) chemoautotroph "Candidatus Brocadia anammoxidans" the interior region bounded by ICM contains a further internal single-membrane bounded region, the anammoxosome. In Gemmata obscuriglobus, the interior ICM bounded region contains the nuclear body, a double-membrane-bounded region containing the cell's nucleoid and all genomic DNA in addition to some RNA. Shared features of cell compartmentalisation in different planctomycetes are consistent with the monophyletic nature of the planctomycetes as a distinct division of the Bacteria. The shared organisational plan for the planctomycete cell constitutes a new type not known in cells of other bacteria. PMID- 11491083 TI - Purification and characterization of a lactonase from Burkholderia sp. R-711, that hydrolyzes (R)-5-oxo-2-tetrahydrofurancarboxylic acid. AB - A lactonase hydrolyzing (R)-5-oxo-2-tetrahydrofurancarboxylic acid to D-alpha hydroxyglutaric acid was purified 170-fold with 2% recovery to near homogeneity from crude extracts of Burkholderia sp. R-711, which had been isolated as a bacterium able to assimilate (R)-5-oxo-2-tetrahydrofurancarboxylic acid. The molecular mass was estimated to be 33 kDa by gel filtration. The purified preparation migrated as a single band of molecular mass 38 kDa upon SDS-PAGE. The maximum activity was observed at pH 7.0-8.0 and 35-40 degrees C. The enzyme required no added cofactors or metal ions; the activity was inhibited to 60-100% by SH-blocking reagents, but was not affected by metal-chelating reagents. The enzyme showed lower activity and affinity toward (S)-5-oxo-2 tetrahydrofurancarboxylic acid, but did not act on other natural and synthetic lactones tested. PMID- 11491084 TI - Rhodococcus sp. RB1 grows in the presence of high nitrate and nitrite concentrations and assimilates nitrate in moderately saline environments. AB - Rhodococcus sp. RB1 was able to thrive in media with up to 0.9 M NaCl or KCl and in the presence of high concentrations of nitrate (up to 0.9 M) and nitrite (up to 60 mM), but only under oxic conditions. An adaptation period was not required for salt tolerance, but a rapid extrusion of K+ and intake of Na+ was observed after addition of 0.5 M NaCl. Nitrate assimilation was limited by the carbon supply, but nitrite was not accumulated in the culture medium, even at nitrate concentrations as high as 0.8 M, thus suggesting that nitrite reduction does not limit nitrate assimilation. The presence of NaCl or KCl did not affect nitrate or nitrite uptake, which were completely inhibited by ammonium or glutamine. Rhodococcus sp. RB1 nitrate reductase had an apparent molecular mass of 142 kDa and used NADH and reduced bromophenol blue or viologens as electron donors, independently of the presence of salt. The enzyme was associated with an NADH diaphorase activity and was induced by nitrate and repressed by ammonium or glutamine, thus showing typical biochemical and regulatory properties of bacterial assimilatory NADH-nitrate reductases. The enzyme was active in vitro in the presence of 3 M NaCl or KCI, but the maximal activity was observed at 0.5 M salt. Addition of 2 M NaCl increased the optimal temperature of the enzyme from 12 to 32 degrees C, but the optimal pH (10.3) was unaffected. PMID- 11491085 TI - Characterization of glucose-repression-resistant mutants of Bacillus subtilis: identification of the glcR gene. AB - In Bacillus subtilis, carbon catabolite repression (CCR) is mediated by the pleiotropic repressor CcpA and by ATP-dependent phosphorylation of the HPr protein of the phosphotransferase system (PTS). In this study, we attempted to identify novel genes that are involved in the signal transduction pathway that ultimately results in CCR in the presence of repressing carbon sources such as glucose. Seven mutants resistant to glucose repression of the levanase operon were isolated and characterized. All mutations were trans-acting and pleiotropic as determined by analyzing CCR of beta-xylosidase and of the sacPA and bglPH operon. Moreover, all mutations specifically affected repression exerted by glucose but not by other sugars. The mutations were mapped to three different loci on the genetic map, ptsG, glcR, and pgi. These three genes encode proteins involved in glucose metabolism. A novel repressor gene, glcR (ywpI), defined by two mutations, was studied in more detail. The glcR mutants exhibit loss of glucose repression of catabolic operons, a deficiency in glucose transport, and absence of expression of the ptsG gene. The mutant GlcR proteins act as super repressors of ptsG expression. PMID- 11491086 TI - Analysis of the stress resistance of commercial wine yeast strains. AB - Alcoholic fermentation is an essential step in wine production that is usually conducted by yeasts belonging to the species Saccharomyces cerevisiae. The ability to carry out vinification is largely influenced by the response of yeast cells to the stress conditions that affect them during this process. In this work, we present a systematic analysis of the resistance of 14 commercial S. cerevisiae wine yeast strains to heat shock, ethanol, oxidative, osmotic and glucose starvation stresses. Significant differences were found between these yeast strains under certain severe conditions, Vitilevure Pris Mouse and Lalvin T73 being the most resistant strains, while Fermiblanc arom SM102 and UCLM S235 were the most sensitive ones. Induction of the expression of the HSP12 and HSP104 genes was analyzed. These genes are reported to be involved in the tolerance to several stress conditions in laboratory yeast strains. Our results indicate that each commercial strain shows a unique pattern of gene expression, and no clear correlation between the induction levels of either gene and stress resistance under the conditions tested was found. However, the increase in mRNA levels in both genes under heat shock indicates that the molecular mechanisms involved in the regulation of their expression by stress function in all of the strains. PMID- 11491087 TI - Purification and characterization of a novel extracellular protease from Bacillus cereus KCTC 3674. AB - Bacillus cereus KCTC 3674 excretes several kinds of extracellular proteases into the growth medium. Two proteases with molecular masses of approximately 36-kDa and 38-kDa, as shown by SDS-PAGE, were purified from the culture broth. The 38 kDa protease was purified from B. cereus cultivated at 37 degrees C, and the 36 kDa protease was obtained from the B. cereus cultivated at 20 degrees C. The 38 kDa protease was identified as an extracellular neutral (metallo-) protease and was further characterized. The 36-kDa protease was shown to be a novel enzyme based on its N-terminal amino acid sequence, its identification as a metallo enzyme that was strongly inhibited by EDTA and o-phenanthroline, its hemolysis properties, and its optimal pH and temperature for activity of 8.0 and 70 degrees C, respectively. PMID- 11491088 TI - Growth of the purple bacterium Rhodobacter capsulatus on the aromatic compound hippurate. AB - The purple nonsulfur bacterium Rhodobacter capsulatus strain B10 grew phototrophically on the aromatic compound hippurate (N-benzoyl-L-glycine) and related benzoyl amino acids. Absorption spectra, extraction, and GC/MS analysis of culture supernatants showed that hippurate was stoichiometrically converted to benzoate and glycine, with the latter used as a carbon or nitrogen source for growth. This conclusion was supported by detection of the enzyme hippuricase in permeabilized intact cells. Chemotrophic growth on hippurate by Rba. capsulatus, either at full or reduced oxygen tensions, was not observed. The type strain of Rhodobacter sphaeroides as well as four strains of Rhodopseudomonas palustris also grew phototrophically on hippurate, while several other aromatic-degrading species of purple bacteria did not. PMID- 11491089 TI - [Diagnosis-related groups]. PMID- 11491090 TI - Safe design and use of panoramic, wet source storage gamma irradiators (category IV) and dry storage gamma irradiators (category II). PMID- 11491091 TI - Vascular endothelial-cell turnover: a new factor in the vascular microenvironment of the liver. PMID- 11491093 TI - 7th Congress of the European Confederation of Medical Mycology (ECMM). Rhodes, Greece, June 16-19, 2001. Abstracts. PMID- 11491092 TI - MHC class II-restricted tumor antigens and CD4+ T cells play a role in hematological malignancies as well as solid tumors. PMID- 11491094 TI - Limited inferior macular translocation for the treatment of subfoveal choroidal neovascularization secondary to age-related macular degeneration. PMID- 11491095 TI - Maximizing surgeon safety during excimer laser photorefractive procedures. PMID- 11491096 TI - Clinicopathology conference: infant with meconium-stained amniotic fluid, poor Apgar scores, hypoxia, and respiratory problems. PMID- 11491098 TI - 23rd Congress of the International Association for Breast Cancer Research. Dusseldorf, Germany, 13-16 June 2001. Abstracts. PMID- 11491097 TI - Biology of chronic lymphocytic leukemia. AB - B-cell chronic lymphocytic leukemia (CLL) lies at the cross-roads of hematology, immunology and oncology for at least three major reasons: (a) it is the prototype of human malignancies that primarily involve defects in the induction of apoptosis; (b) CLL patients develop a severe immunodeficiency with progressive hypogammaglobulinemia; and (c) they have a high prevalence of autoimmune phenomena. Recent advances in the biology of the malignant cell in CLL lead to a scenario comprised of two basic elements: first, CLL cells are optimally organized to survive in their niches because their ability to undergo apoptosis is severely hampered; second, they have a microenvironment-dependence that promotes their extended survival, a situation that arises most probably through direct cell-to-cell contacts. In addition, CLL cells themselves are the major accessory cells in CLL, but are inefficient antigen-presenting cells. This latter defect may provide a clue to reinterpret the events of immunodeficiency and autoimmunity. PMID- 11491099 TI - India backtracks on Bt cotton. PMID- 11491100 TI - MorphoSys, CAT fight. PMID- 11491101 TI - Confirmant to sell protein data. PMID- 11491102 TI - BCG report rosy on genomics. PMID- 11491103 TI - Haplotype map planned. PMID- 11491104 TI - New WTO database? PMID- 11491105 TI - NASD rule requires disclosure. PMID- 11491106 TI - ESF endorses therapeutic cloning. PMID- 11491107 TI - Off air benzene degradation. PMID- 11491108 TI - ES cells leave their imprint. PMID- 11491109 TI - Piezoelectric arrays. PMID- 11491110 TI - ECM 2000. Abstracts of an International Conference on the Biology and Pathology of the Extracellular Matrix. St. Louis, Missouri, USA. October 12-15, 2000. PMID- 11491112 TI - [53rd Congress of the German Association of Urology. 19-22 September 2001, Dusseldorf. Abstracts]. PMID- 11491114 TI - Abstracts of the 24th annual meeting of the American Society of Primatologists. August 8-11, 2001. Savannah, Georgia, USA. PMID- 11491113 TI - Abstracts of the 11th Conference of the European Society for Psychosocial Oncology. June 13-16, 2001. Heidelberg, Germany. PMID- 11491115 TI - An ancient cluster of Wnt paralogues. PMID- 11491116 TI - Minocycline and Pseudotumor cerebri: The well-known but well-kept secret. PMID- 11491117 TI - Occult bacteremia from a pediatric emergency department. PMID- 11491118 TI - Back to sleep: good advice for parents but not for hospitals? PMID- 11491119 TI - Circumcisions: again. PMID- 11491120 TI - Barotrauma, oxygen toxicity, and chronic lung disease. PMID- 11491121 TI - Pacifier use is associated with shorter breastfeeding duration among low-income women. PMID- 11491122 TI - A sudden rise in the prevalence of retinopathy of prematurity blindness? PMID- 11491124 TI - American Association of Oral and Maxillofacial Surgeons 83rd annual meeting. September 12-15, 2001. Orlando, Florida, USA. Abstracts. PMID- 11491123 TI - ISBT VII European Congress. Paris, France, July 15-18, 2001. Abstracts. PMID- 11491125 TI - The health effects of tea and tea components. PMID- 11491126 TI - American Society for Mass Spectrometry directory of members and corporate members. PMID- 11491127 TI - Abstracts from the 26th Congress of the Scandinavian Society of Anaesthesiology and Intensive Care Medicine. June 13-17, 2001. Tromso, Norway. PMID- 11491128 TI - Anomalous pH dependence of the reactions of carbenicillin and sulbenicillin with Bacillus cereus beta-lactamase I. Influence of the alpha-substituent charge on the kinetic parameters. AB - The pH dependence of k(cat) for the Bacillus cereus beta-lactamase I catalyzed hydrolysis of carbenicillin(VI), which differs from benzylpenicillin (I) in having a carboxylic moiety alpha to the phenyl ring, exhibits a profile consistent with a model in which the alpha-COOH and alpha-COO forms of the ES complex turn over with respective rate constants of 2152 s(-1) and 384 s(-1). The pK(a)(app) for the alpha-COOH is shifted from 3.2 in solution to 6.1 in the ES complex. The normalized k(cat)/K(m) vs. pH profile for VI is not superimposable on that of I, indicating that both the neutral and anionic forms of the carboxyl moiety of VI combine with the enzyme to give the first irreversibly formed complex, presumably the acyl-enzyme. Quantitative accord with the kinetic data is achieved only through fitting to a model where kinetically significant proton transfer in the ES complex is permitted. The second-order rate constants for the reaction of the enzyme with the alpha-COOH and alpha-COO forms of VI are 2.2 x 10(8) M(-1) s(-1) and 3.8 x 10(6) M(-1) s(-1), respectively. The high value for the alpha-COOH form suggests that this reaction may be in part diffusion controlled. This conjecture is borne out by the observation that the sensitivity of k(cat)/K(m) to eta(rel) decreases with increasing pH for VI, whereas this sensitivity is pH independent for I. These conclusions are further supported by the results of a kinetic investigation of the pH dependence of sulbenicillin (VII) where an alpha-SO3H replaces the alpha-COOH of VI. The strongly acidic sulfonic acid moiety of VII is fully ionized throughout nearly the entire pH range of interest, and its kinetics, as a function of pH, are very similar to those observed and calculated for the alpha-COO form of VI. Solvent deuterium kinetic isotope effects are reported for k(cat) and k(cat)/K(m) for both VI and VII. PMID- 11491129 TI - Diffusion-limited component of reactions catalyzed by Bacillus cereus beta lactamase I. AB - The Bacillus cereus beta-lactamase I catalyzes the hydrolysis of a wide variety of penicillins and cephalosporins with values of k(cat)/K(m) varying over several orders of magnitude. The values of this parameter for the most reactive of these compounds, benzylpenicillin, I, and furylacryloyl-penicillin, II (k(cat)/K(m) = 2.43 x 10(7) M(-1) s(-1) and 2.35 x 10(7) M(-1) s(-1), respectively, at pH 7.0 in potassium phosphate buffer containing 0.17 M KCl, I(c) = 0.63, 25 degrees C) are decreased markedly by increasing viscosity in sucrose- or glycerol-containing buffers. The relative sensitivities to viscosity of k(cat)/K(m) values for I and for cephaloridine, III, were found to be virtually unchanged at pH 3.8 from those observed at pH 7.0. The differential effects of viscosity on the reactive vs. the sluggish [e.g., cephalothin (IV), k(cat)/K(m) = 1 x 10(4) M(-1) s(-1)] substrates support the contention that the rates of reaction of the former with the enzyme are in part diffusion controlled. Quantitative analysis gives values for the association rate constants, k(1), of 7.6 x 10(7) M(-1) s(-1), 4 x 10(7) M(-1) s( 1), and 1.1 x 10(7) M(-1) s(-1) for I, II, and III, respectively. As both reactive and sluggish substrates associate with the active site of the enzyme with relatively similar rate constants, the variation in k(cat)/K(m) values is primarily due to the variation in the partition ratios k(-1)/k(2), for the ES complex, which are 2.3, 0.77, and 30 for I, II, and III, respectively. The preceding analysis is based on direct application of the Stokes-Einstein diffusion law to enzyme kinetics. The range of applicability of this law to the diffusion of substrate size molecules and the mechanics of diffusion of ionic species through viscous solutions of sucrose vs. polymers are explored. PMID- 11491130 TI - A simple provider-based educational intervention to boost infant immunization rates: a controlled trial. AB - We sought to determine if a simple educational intervention initiated at the first well-child care visit, with reinforcement at subsequent visits, can improve inner-city infant immunization rates. We conducted a controlled trial involving 315 newborn infants and their primary caregivers in 3 inner-city primary care centers. Child health care providers gave caregivers in the intervention group an interactive graphic card with verbal reinforcement. At later visits, stickers were applied to the card when immunizations were given. Routine information was given to controls. After the trial, age-appropriate immunization rates at 7 months were 58% in each group. Intervention infants had 50% fewer missed opportunities to immunize (p=0.01) but cancelled 77% more appointments (p=0.04) than controls. We conclude that a brief educational intervention at the first well-child care visit did not boost 7-month immunization rates, although it was associated with fewer missed opportunities to immunize. PMID- 11491132 TI - Factors in head injury in children. PMID- 11491131 TI - Association among duration of unconsciousness, Glasgow Coma Scale, and cranial computed tomography abnormalities in head-injured children. AB - The impact of head trauma leads to generation of forces that cause both the intracranial injuries and loss of consciousness. Glasgow Coma Score (GCS) and cranial CT scan are widely used to gauge the severity of head injury. Milder cranial impacts insufficient to cause intracranial injuries generally cause no or minimal loss of consciousness. Nevertheless, cases with contradictory findings are also seen. This study was undertaken to find out the association among the three above-stated variables. A knowledge of such an association can enable the attending clinician in prognosticating a cranial injury and selecting out those patients with mild head injuries who deserve closer observation. While a significant positive association was found between duration of unconsciousness and GCS, no signifficant association of either of these variables with CT scan findings was noted. PMID- 11491133 TI - A randomized, blinded comparison of chloral hydrate and midazolam sedation in children undergoing echocardiography. AB - The objective of this prospective, randomized, and blinded study was to compare the use of chloral hydrate versus oral midazolam sedation in children undergoing echocardiography. No adverse effects (nausea, vomiting, paradoxical agitation, or significant deviations from baseline vital signs) were noted with either medication. No differences were noted in onset of sedation between the 2 groups, however, the time to complete recovery was significantly shorter with midazolam than with chloral hydrate. The children in the chloral hydrate group had a significantly deeper level of sedation and were more likely to receive a more nearly comprehensive echocardiographic evalation. PMID- 11491134 TI - Antacid-induced rickets in infancy. AB - A 3-month-old premature infant presented with a "soft skull." Clinical and radiologic findings confirmed the diagnosis of rickets. Biochemistry revealed normal serum parathyroid hormone (PTH) and undetectable urine phosphate. These findings combined with a history of 5-6 weeks' treatment with high-dose aluminum rich antacid established the diagnosis of antacid-induced rickets. Discontinuation of the medicine combined with phosphate and vitamin D supplementation resulted in quick resolution of all clinical, radiologic, and biochemical abnormalities. Our patient demonstrates that in premature infants antacid-induced rickets can develop within a few weeks; normal serum PTH concentration and hypophosphaturia are highly indicative of the diagnosis, and contrary to the situation in adults in whom hypercalciuria has been often described, in infants hypocalciuria is more commonly observed. Pediatricians should avoid or minimize the use of aluminum-containing antacids, and when used, carefully monitor mineral metabolism. PMID- 11491135 TI - An unusual cause of a red and swollen eye in an 11-month-old child. PMID- 11491136 TI - Allergic constipation: association with infantile milk allergy. PMID- 11491137 TI - Lipoid congenital adrenal hyperplasia (CAH): patient report and a mini-review. PMID- 11491138 TI - A training model for circumcision of the newborn. PMID- 11491139 TI - Reversible subacute combined degeneration of the spinal cord in a 14-year-old due to a strict vegan diet. PMID- 11491140 TI - A case of idiopathic gynecomastia treated by endoscopic reduction mammoplasty. PMID- 11491141 TI - Cutis laxa with growth and developmental delay. PMID- 11491142 TI - Bone mineral density in adolescents. PMID- 11491143 TI - Do long-acting beta2-adrenoceptor agonists enhance the anti-inflammatory effect of glucocorticoids in asthma? PMID- 11491144 TI - Chest computed tomography: is it ready for major studies of chronic obstructive pulmonary disease? PMID- 11491145 TI - Severe pulmonary hypertension after the discovery of the familial primary pulmonary hypertension gene. AB - The recent discoveries of the familial primary pulmonary hypertension gene and somatic mutations in key cell growth and cell death regulatory genes in primary pulmonary hypertension have added a new dimension to severe pulmonary hypertension research. These findings have already impacted on how the disease is viewed, and ultimately, how severe pulmonary hypertension is diagnosed and treated. However, this new information raises several fundamental questions related to the role of bone morphogenetic protein receptor signalling in the control of lung vascular cell function. Furthermore, additional genes and gene products may also be involved in the pathogenesis of the disease. The way severe pulmonary hypertension is viewed and studied is on the verge of shifting from a vasoconstrictive to a cell growth paradigm. PMID- 11491146 TI - Effects of formoterol and budesonide on GM-CSF and IL-8 secretion by triggered human bronchial epithelial cells. AB - The effect of formoterol, alone and in combination with budesonide, upon tumour necrosis factor-alpha stimulated (10 ng x mL(-1)) human bronchial epithelial cells was investigated. Addition of formoterol (> or = 10(-10) M) reduced granulocyte macrophage-colony stimulating factor (GM-CSF) levels, as assessed by enzyme-linked immunosorbent assay, by 40-50% and increased interleukin (IL)-8 levels by approximately 50%. The effects of formoterol were long lasting (23 h). Budesonide (10(-8) M) reduced the amounts of both cytokines (GM-CSF and IL-8) by 40%. Simultaneous addition of formoterol and budesonide reduced GM-CSF levels approximately 75%, while IL-8 levels were decreased approximately 40%, similar to the reduction obtained with budesonide alone. The glucocorticoid receptor (GR) antagonist RU486 did not influence the effect of formoterol, suggesting no involvement of the GR. Formoterol rapidly induced an elevation in intracellular cyclic adenosine monophosphate, which was reduced in the presence of propranolol. In addition, the alterations in cytokine secretion induced by formoterol could be fully blocked by propranolol, demonstrating that these effects are beta2-receptor mediated. In conclusion, the combination of budesonide and formoterol reduces the secretion of granulocyte macrophage-colony stimulating factor to basal levels and counteracts the capacity of formoterol alone to induce interleukin-8 production, modulations which may facilitate improved asthma control. PMID- 11491147 TI - Host immune response to Chlamydia pneumoniae heat shock protein 60 is associated with asthma. AB - Chlamydia pneumoniae infection has been associated with asthma. It has also been suggested that heat shock protein 60 (Hsp60) belonging to a class of highly conserved proteins may play a role in the pathogenesis of chlamydial infections. The purpose was to study whether the host immune response to C. pneumoniae Hsp60 is associated with asthma and decreased pulmonary function. An enzyme immunoassay was used to measure immunoglobulin-(Ig)A and IgG antibodies against recombinant C. pneumoniae Hsp60 and human Hsp60 in a study group consisting of 24 cases of recently symptomatic asthma and 62 nonasthmatic controls. A strong (r=0.50) and significant (p<0.001) correlation was observed between C. pneumoniae and human Hsp60 IgA antibodies, but only C. pneumoniae Hsp60 IgA antibodies were significantly associated with asthma (p = 0.02). Pulmonary function, as measured by forced expiratory volume in one second, also inversely correlated (r = -0.23, p = 0.04) with the quantity of C. pneumoniae Hsp60 IgA antibodies, suggesting an association with the severity of pulmonary obstruction. By showing an association of Chlamydia pneumoniae heat shock protein 60 immunoglobulin A antibodies with asthma, the results support the hypothesis of an association between Chlamydia pneumoniae infection and asthma and support the need for further investigations on the role of heat shock protein 60 in the pathogenesis of asthma. PMID- 11491148 TI - Treatment of asthma by the inhaled corticosteroid ciclesonide given either in the morning or evening. AB - The study addressed the question whether the novel inhaled prodrug corticosteroid ciclesonide is equally effective when inhaled in the morning compared to the evening. For this purpose a double-blind, randomized, parallel group study was initiated in which 209 asthmatic patients (forced expiratory volume in one second = 50-90% predicted) inhaled either 200 microg ciclesonide in the morning or in the evening, for 8 weeks. Efficacy was assessed by means of spirometry as well as daily recordings of morning and evening peak expiratory flow (PEF), symptoms and use of rescue medication. The 24-h urinary cortisol excretion was measured to evaluate any effect on hypothalamic-pituitary-adrenol axis. Ciclesonide significantly improved asthma control. Morning and evening administration was shown to be equally effective for the different spirometry variables, evening PEF, symptoms, use of rescue medication and number of asthma exacerbations. Regarding morning PEF, the improvements after evening dosing were more prominent and equivalence of morning and evening administration could not be demonstrated. No relevant influence on cortisol excretion was found. Overall, the study indicates that ciclesonide can be given either in the morning or in the evening to meet the patients' preference and individual medical needs, although evening administration may lead to a more pronounced improvement in morning peak expiratory flow. PMID- 11491149 TI - High prevalence of asthma in five remote indigenous communities in Australia. AB - Data on the prevalence of asthma in children residing in remote indigenous communities in Australia are sparse, despite the many reports of high prevalence in nonindigenous children of this country. Two previous Australian studies have had poor participation rates, limiting interpretation of their results. A study of children in the Torres Strait and Northern Peninsula Area of Australia was conducted to document the prevalence of asthma symptoms. Five indigenous communities were randomly selected and trained interviewers, who were local indigenous health workers, recruited participants using a house-by-house approach. Information was collected by a structured face-to-face interview based on a standardized questionnaire constructed from the protocol International Study of Asthma and Allergy in Childhood; 1,650 children were included in the study with a 98% response rate. Overall, the prevalence of self-reported ever wheezing was 21%; 12% reported wheezing in the previous year; and 16% reported ever having asthma. There was significant variation in the prevalence of asthma symptoms between communities. It is concluded that there are significant intercommunity variations in the prevalence of asthma symptoms in remote communities and that the prevalence in these communities is as high as in nonindigenous groups. PMID- 11491150 TI - High-resolution computed tomography scanning in alpha1-antitrypsin deficiency: relationship to lung function and health status. AB - The development of computed tomography (CT) has enabled emphysema to be assessed noninvasively. Objective quantification of lung density correlates well with lung function in patients with chronic obstructive pulmonary disease and has been shown to be a sensitive tool for monitoring disease progression. In order to determine the clinical impact of changes seen on high-resolution computed tomography (HRCT), the relationship between the objective quantification of emphysema on HRCT, lung function and health status in 111 patients with alpha1 antitrypsin deficiency was examined (PiZ). The degree of HRCT scan abnormality correlated well (p<0.001 for all comparisons) with forced expiratory volume in one second (r = -0.60- -0.75), specific airway conductance (r = -0.67-0.76), residual volume/total lung capacity (r = 0.46-0.58) and transfer factor of the lung for carbon monoxide (r = -0.64- -0.81). In addition, the CT scans correlated (p<0.001) with health status as assessed by the St. George's Respiratory Questionnaire (SGRQ total: r = -0.38-0.50) and the Short-Form health survey (e.g. physical functioning: r = -0.39-0.54). In summary, other workers have shown high resolution computed tomography to be a sensitive indicator of disease progression. This study confirms the relationship between high-resolution computed tomography and lung physiology, and suggests the relationship is even stronger in patients with predominantly lower zone pan-lobular emphysema than in usual chronic obstructive pulmonary disease. High-resolution computed tomography also relates to patients disability and impairment as defined by health status questionnaires and, therefore, should be considered as an alternative outcome measure particularly in alpha1-antitrypsin deficiency. PMID- 11491151 TI - Increased circulating levels of alpha1-antitrypsin and calprotectin are associated with reduced gas diffusion in the lungs. AB - The aim was to examine the relationship of serum inflammatory markers to the level of single-breath diffusing capacity of carbon monoxide (TL,CO). A stratified sample (n = 1,121) of a Norwegian general population aged 18-73 yrs was examined. The inflammatory markers measured were calprotectin, a prominent protein in the cytosol fraction of neutrophil granulocytes, and alpha1 antitrypsin (alpha1-AT), the major inhibitor of neutrophil elastase in the lower respiratory system. Both markers have increased circulating levels in the course of an acute inflammatory reaction. Subjects with a TL,CO < 80% of predicted value had a higher level of both alpha1-AT (p = 0.003) and calprotectin (p < 0.03) than those with a TL,CO > 100%. In multiple linear regression analyses, alpha1-AT was still significantly associated with TL,CO after adjusting for sex, age, smoking habits, haemoglobin, carboxyhaemoglobin, forced expiratory volume in one second and alveolar volume. In a similar analysis, no significant overall association was found between calprotectin and TL,CO, but in a stratified analysis, calprotectin was significantly related to TL,CO in females. However, no significant sex interaction in the relationship between the inflammatory markers and TL,COo was found. The findings suggest that increased levels of alpha1 antitrypsin and of calprotectin are risk factors for decreased diffusing capacity of carbon monoxide. PMID- 11491152 TI - Changes in bronchial inflammation during acute exacerbations of chronic bronchitis. AB - There are little data describing noncellular changes in bronchial inflammation during exacerbations of chronic bronchitis. The relationship between sputum colour and airway inflammation at presentation has been assessed during an exacerbation in patients with chronic bronchitis and a primary care diagnosis of chronic obstructive pulmonary disease. Sputum myeloperoxidase, neutrophil elastase, leukotriene B4 (LTB4), interleukin-8 (IL-8), sol:serum albumin ratio and serum C-reactive protein were measured in patients presenting with an exacerbation and mucoid (n = 27) or purulent sputum (n = 42). Mucoid exacerbations were associated with little bronchial or systemic inflammation at presentation, and sputum bacteriology was similar to that obtained in the stable state. Purulent exacerbations were associated with marked bronchial and systemic inflammation (p < 0.025 for all features) and positive sputum cultures (90%). Resolution was related to a significant reduction in LTB4 (p < 0.01), but no change in IL-8, suggesting that LTB4 may be more important in neutrophil recruitment in these mild, purulent exacerbations. In the stable state, IL-8 remained higher in patients who had experienced a purulent exacerbation (2p < 0.02). The presented results indicate that exacerbations of chronic bronchitis, defined by sputum colour, differ in the degree of bronchial and systemic inflammation. Purulent exacerbations are related to bacterial infection, and are associated with increased neutrophilic inflammation and increased leukotriene B4 concentrations. PMID- 11491153 TI - Breathing pattern and gas exchange at peak exercise in COPD patients with and without tidal flow limitation at rest. AB - Expiratory flow limitation (FL) at rest is frequently present in chronic obstructive pulmonary disease (COPD) patients. It promotes dynamic hyperinflation with a consequent decrease in inspiratory capacity (IC). Since in COPD resting IC is strongly correlated with exercise tolerance, this study hypothesized that this is due to limitation of the maximal tidal volume (VT,max) during exercise by the reduced IC. The present study investigated the role of tidal FL at rest on: 1) the relationship of resting IC to VT,max; and 2) on gas exchange during peak exercise in COPD patients. Fifty-two stable COPD patients were studied at rest, using the negative expiratory pressure technique to assess the presence of FL, and during incremental symptom-limited cycling exercise to evaluate exercise performance. At rest, FL was present in 29 patients. In the 52 patients, a close relationship of VT,max to IC was found using non-normalized values (r=0.77; p < 0.0001), and stepwise regression analysis selected IC as the only significant predictor of VT,max. Subgroup analysis showed that this was also the case for patients both with and without FL (r=0.70 and 0.76, respectively). In addition, in FL patients there was an increase (p < 0.002) in arterial carbon dioxide partial pressure at peak exercise, mainly due to a relatively low VT,max and consequent increase in the physiological dead space (VD)/VT ratio. The arterial oxygen partial pressure also decreased at peak exercise in the FL patients (p < 0.05). In conclusion, in chronic obstructive pulmonary disease patients the maximal tidal volume, and hence maximal oxygen consumption, are closely related to the reduced inspiratory capacity. The flow limited patients also exhibit a significant increase in arterial carbon dioxide partial pressure and a decrease in arterial oxygen partial pressure during peak exercise. PMID- 11491154 TI - Usefulness of transcutaneous Doppler jugular venous echo to predict pulmonary hypertension in COPD patients. AB - Pulmonary hypertension is an important factor that determines the prognosis of chronic obstructive pulmonary disease (COPD) patients. Echocardiography is a noninvasive and useful bedside method for measurement of pulmonary artery pressure. However, this method is sometimes difficult because of the overinflated lungs in COPD patients. This study attempted to estimate pulmonary hypertension in COPD patients using transcutaneous Doppler jugular vein flow velocity recording. The mean pulmonary artery pressure (MPAP) of 64 COPD patients was examined using cardiac catheterization. The right jugular vein flow velocity was measured within 24 h using transcutaneous Doppler echo, after which the ratio of diastolic flow (Df) and systemic flow (Sf) velocity was calculated. Subsequently, the statistical correlation of MPAP and the Df/Sf ratio was examined. MPAP was also measured using standard cardiac echo methods and the results were compared. The Df/Sf velocity ratio showed significant correlation with MPAP in COPD patients (r=0.844, p<0.0001). The sensitivity was 71.4%, and the specificity 95.3% (cut-off ratio= 1.0). Jugular venous Doppler echo could be performed in all patients while other cardiac echo methods could not be performed in all patients. The specificity of the methods used was higher than other cardiac echo methods. Transcutaneous jugular vein flow velocity measurement may be applicable to bedside prediction of pulmonary hypertension in chronic obstructive pulmonary disease patients. PMID- 11491155 TI - Effectiveness of salmeterol versus ipratropium bromide on exertional dyspnoea in COPD. AB - The hypothesis of the study was that salmeterol and ipratropium would have similar dyspnoea ratings during steady-state cycle ergometry at 1 h, but that salmeterol would reduce dyspnoea at 6 h after administration in patients with chronic obstructive pulmonary disease (COPD). The study design was a randomized, double-blind trial in 16 patients (aged 63 +/- 11 yrs) with symptomatic COPD. Two days after familiarization with testing procedures, patients were randomly assigned to receive either two puffs (42 microg) of salmeterol and two puffs of placebo inhaler, or two puffs (36 microg) of ipratropium from each of two inhalers (total, 72 microg). Two days later, patients received the alternative medication. During exercise at 60% of peak oxygen consumption patients rated dyspnoea and performed inspiratory capacity manoeuvres each minute. Forced expiratory volume in one second was 1.13+/-0.48 L (37+/-13% predicted). Dyspnoea ratings were similar for salmeterol and ipratropium at 1 and 6 h. Inspiratory capacity was similar for salmeterol and ipratropium at 1 h, but significantly higher for salmeterol at 6 h (delta = 120 mL; p = 0.03). It is concluded that with the doses used, salmeterol and ipratropium provided similar dyspnoea ratings during exercise at 1 and 6 h after administration. PMID- 11491156 TI - Supported discharge shortens hospital stay in patients hospitalized because of an exacerbation of COPD. AB - This prospective, controlled, but not formally randomized study investigates the feasibility and efficiency of an alternative to standard hospitalization for patients with exacerbated chronic obstructive pulmonary disease (COPD), based upon supported discharge with nurse supervision at home. Over a 12-month period, emergency physicians, not directly involved in the study, admitted 205 patients with exacerbated COPD to the authors' respiratory unit. Patients were included in the supported discharge group (n=105) if they voluntarily chose to participate in the programme and lived in the city of Palma de Mallorca (where adequate home support could be provided). Patients not fulfilling these criteria (mainly residents outside the city) served as controls (n=100). Inpatient treatment was standardized in all patients and included oxygen therapy, bronchodilators, antibiotics and steroids. Both groups were comparable in terms of age (mean +/- SD: 70 +/- 10 versus 65 +/- 11 yr for supported discharge and control group, respectively), severity of airflow obstruction (forced expiratory volume in one second 45 +/- 18% reference versus 46 +/- 19% ref.), comorbidity and socioeconomic status. Length of hospital stay (LOS) in the supported discharge group was shorter (5.9 +/- 2.8 versus 8.0 +/- 5.1 days, p < 0.001). After discharge, a respiratory nurse visited supported discharge patients at home during 7.3 +/- 3.8 days. Only one patient (1%) required hospital readmission during this period of time. The reduced LOS resulted in a lower utilization of hospital beds at any given point in time throughout the study period. Within the framework and potential limitations of this study, the results indicate that the supported discharge programme in Spain: 1) allows a significant reduction in the length of hospital stay of patients hospitalized because of an exacerbation of chronic obstructive pulmonary disease; 2) does not result in an inappropriately increased rate of hospital readmissions; and 3) reduces the utilization of hospital resources. PMID- 11491157 TI - Air pollution and hospital admissions for respiratory conditions in Rome, Italy. AB - Most of the evidence regarding the association between particulate air pollution and emergency room visits or hospital admissions for respiratory conditions and asthma comes from the USA. European time-series analyses have suggested that gaseous air pollutants are important determinants of acute hospitalization for respiratory conditions, at least as important as particulate mass. The association between daily mean levels of suspended particles and gaseous pollutants (sulphur dioxide, nitrogen dioxide, carbon monoxide, ozone) was examined. The daily emergency hospital admissions for respiratory conditions in the metropolitan area of Rome during 1995-1997 were also recorded. Daily counts of hospital admissions for total respiratory conditions (43 admissions day(-1)), acute respiratory infections including pneumonia (18 day(-1)), chronic obstructive pulmonary disease (COPD) (13 day(-1)), and asthma (4.5 day(-1)) among residents of all ages and among children (0-14 yrs) were analysed. The generalized additive models included spline smooth functions of the day of study, mean temperature, mean humidity, influenza epidemics, and indicator variables for day of the week and holidays. Total respiratory admissions were significantly associated with same-day level of NO2 (2.5% increase per interquartile range (IQR) change, 22.3 microg x m(-3)) and CO (2.8% increase per IQR, 1.5 mg x m( 3)). No effect was found for particulate matter and SO2, whereas O3 was associated with admissions only among children (lag 1, 5.5% increase per IQR, 23.9 microg x m3). The effect of NO2 was stronger on acute respiratory infections (lag 0, 4.0% increase) and on asthma among children (lag 1, 10.7% increase). The admissions for all ages for asthma and COPD were associated only with same-day level of CO (5.5% and 4.3% increase, respectively). Multipollutant models confirmed the role of CO on all respiratory admissions, including asthma and COPD, and that of NO2 on acute respiratory infections. Among children, O3 remained a strong indicator of acute respiratory infections. Carbon monoxide and photochemical pollutants (nitrogen dioxide, ozone) appear to be determinants of acute respiratory conditions in Rome. Since carbon monoxide and nitrogen dioxide are good indicators of combustion products from traffic related sources, the detected effect may be due to unmeasured fine and ultrafine particles. PMID- 11491158 TI - Fewer allergic respiratory disorders among farmers' children in a closed birth cohort from Sweden. AB - The aim of this study was to investigate the prevalence of respiratory allergy, eczema and atopic sensitization in a closed birth cohort of Swedish schoolchildren, 7-8 yrs of age (n=707), of farmers and nonfarmers on the island of Gotland, in the Baltic Sea. All children were born and raised on the island. The survey comprised a questionnaire on atopic diseases and lifestyle factors. Atopic sensitization was assessed by the skin-prick test (SPT) with 15 standardized allergens. The risk ratio (RR) for ever having asthma and/or allergic rhinoconjunctivitis was significantly lower among children of farmers compared to children of nonfarmers (RR=0.38, confidence interval (CI) 95% 0.19 0.77). SPTs (test rate 92%) showed that 32% of the children had at least one positive test. Although the number of positive SPTs did not differ between the groups, there was a reduced risk among children of farmers for having both respiratory symptoms and sensitization to any International Study on Asthma and Allergy in Childhood allergen (RR=0.28, CI 95% 0.09-0.88). The present indicate that living in a farming population seems to protect against development of respiratory allergic disorders but not against allergic sensitization. PMID- 11491159 TI - Monitoring of nonlinear respiratory elastance using a multiple linear regression analysis. AB - The elastic pressure/volume (P/V) curve obtained by the multiple linear regression (MLR) technique using a new model, was compared with the quasi-static P/V points obtained by the rapid airway occlusion technique. Seven infants were studied during mechanical ventilation using a pressure controlled mode. The resistive pressure was subtracted from airway opening pressure, thus determining the elastance related pressure, which was then plotted against the volume to make an MLR-elastance curve. Quasi-static P/V curves of the rapid occlusion technique were constructed by plotting the different inspiratory and expiratory volumes against the corresponding values of the quasi-static airway pressure. The calculated MLR-elastance curves closely fit the experimental quasi-static P/V points obtained by the occlusion technique. There were, however, some discrepancies due to the viscoelastic behaviour of the respiratory system. Although slightly altered by these discrepancies, the multiple linear regression elastance curves did fit the observed quasi-static pressure/volume characteristics for use in clinical practice. The multiple linear regression technique may prove to be clinically useful by continuous monitoring of respiratory system mechanics during mechanical ventilation. PMID- 11491160 TI - Noninvasive measurement of mean alveolar carbon dioxide tension and Bohr's dead space during tidal breathing. AB - The lack of methodology for measuring the alveolar carbon dioxide tension (PA,CO2) has forced investigators to make several assumptions, such as that PA,CO2 is equal to end-tidal (PET,CO2) and arterial CO2 tension (Pa,CO2). The present study measured the mean PA,CO2 and Bohr's dead space ratio (Bohr's dead space/tidal volume (VD,Bohr/VT)) during tidal breathing. The method used is a new, simple and noninvasive technique, based on the analysis of the expired CO2 volume per breath (VCO2) versus the exhaled VT. This curve was analysed in 21 normal, healthy subjects and 35 chronic obstructive pulmonary disease (COPD) patients breathing tidally through a mouthpiece apparatus in the sitting position. It is shown that: 1) PA,CO2 is similar to Pa,CO2 in normal subjects, whilst it is significantly lower than Pa,CO2 in COPD patients; 2) PA,CO2 is significantly higher than PET,CO2 in all subjects, especially in COPD patients; 3) VD,Bohr/VT is increased in COPD patients as compared to normal subjects; and 4) VD,Bohr/VT is lower than the "physiological" dead space ratio (VD,phys/VT) in COPD patients. It is concluded that the expired carbon dioxide versus tidal volume curve is a useful tool for research and clinical work, because it permits the noninvasive and accurate measurement of Bohr's dead space and mean alveolar carbon dioxide tension accurately during spontaneous breathing. PMID- 11491161 TI - A bibliometric evaluation of European Union research of the respiratory system from 1987-1998. AB - This study analyses the evolution of the bibliometric indicators of productivity and repercussion of European Union (EU) research into the respiratory system during the period from 1987-1998, describing the geographical distribution. Using MedLine, a selection was made of those articles by EU authors published between 1987-1998 in 38 respiratory system journals (classification from the Institute for Scientific Information). The journals, country of origin, number of articles and the relation to socioeconomic data, productivity index, visibility index, expected impact factor (EIF) and relative impact factor (RIF) were all analysed. The number of EU publications in respiratory system journals experienced an exponential increase, going from 606 articles (14.3% of world production) in 1987, to 2,325 (33.2%) in 1998. During this same period, the EIF increased from 1,258 to 2,111. The greatest gross productivities were those of the UK, France, Italy and Germany, although when corrected for number of inhabitants, Sweden, the Netherlands, Belgium and Denmark headed the list. The countries with the greatest mean EIF were the Netherlands, the UK, Spain and Belgium. In conclusion, productivity and repercussions of European Union research of the respiratory system experienced an important increase during this period. PMID- 11491162 TI - High morbidity and mortality in cystic fibrosis patients compound heterozygous for 3905insT and deltaF508. AB - Genotype-phenotype association in cystic fibrosis (CF) is difficult because of heterogeneous disease expression. The genotype-phenotype correlation for the 3905insT mutation in comparison to deltaF508 was studied here. Thirty CF patients compound heterozygous for 3905insT were compared to clinical presentation of matched patients homozygous for deltaF508 (1960-1997). Sweat tests, age at diagnosis, at death and at onset of Pseudomonas aeruginosa colonization were analysed. Chrispin-Norman scores and pulmonary function forced expiratory volume in one second (FEV1) determined severity of lung disease. Twenty-five of the patients with 3905insT had deltaF508 as a second mutation and five had another rare mutation. At the age of 15 yrs, 60% of patients with 3905insT had an FEV1 < 60% predicted in comparison to 25% of patients with deltaF508 (p<0.05). Age at death and cumulative survival rate was significantly lower (p<0.05) in the 3905insT than in the deltaF508 group (20.3 and 24.0 yrs, respectively). Age at onset of P. aeruginosa colonization was not different in the study groups. Sweat chloride concentrations were lower in patients homozygous for deltaF508 (105.63+/ 15.3 mmol L(-1)) than in patients with 3905insT (119.9+/-22.1 mmol x L(-1)) (p<0.05). Patients compound heterozygous for 3905insT have similar high morbidity and mortality to patients homozygous for deltaF508. PMID- 11491163 TI - The German cystic fibrosis quality assurance project: clinical features in children and adults. AB - Cystic fibrosis (CF) is a complex disease which requires interdisciplinary care in specialized CF centres. In Germany, 97 paediatric and adult outpatient clinics agreed to report clinical data of their patients to a newly established registry, the Cystic Fibrosis Quality Assurance (CFQA) project. This article characterizes the design of the CFQA and the health status of the patients enrolled by the end of 1997. Data from 4,306 patients reported to the CFQA project were analysed. Nutritional status and lung function of the patients were examined as well as the use of specific therapeutic interventions. Mean age of all 4,182 patients alive by the end of 1997 was 15.7 yrs (maximum, 58 yrs), and 35.8% of patients were > 18.0 yrs of age. One-third of the CF population were treated in the nine largest centres (each caring for > 100 patients). Abnormal nutritional status (weight-for height >90% of predicted or body mass index < 19.0 kg m2, respectively) was observed in 26.8% of children and adolescents and in 38.3% of adults. Lung function was abnormal (forced expiratory volume in one second < 80% predicted) in the majority of adults (83.9%) and in 42.5% of the younger patients. The mortality rate was 1.4 of 100 patients in 1997. No clear association of clinical status with centre size was observed. The clinical features of patients treated in German cystic fibrosis centres were generally comparable to those reported from other countries, although improvements are certainly warranted. The Cystic Fibrosis Quality Assurance project represents an important tool for future progress in the quality of cystic fibrosis care. PMID- 11491164 TI - Intron-8 polythymidine sequence in Australasian individuals with CF mutations R117H and R117C. AB - Compound heterozygotes for a severe cystic fibrosis transmembrane conductance regulator (CFTR) mutation and the R117H or R117C mutation (R117H/C) have clinical presentations that vary from classic cystic fibrosis (CF) to an incidental genetic finding. The aim of this study was to assess the influence of the intron 8 polythvmidine sequence (IVS8) on the relationship between genotype and phenotype of individuals with R117H/C. All individuals with R117H/C known to CF clinics in Australia and New Zealand were retrospectively studied by collecting information on genotype, age, pancreatic status, sweat electrolytes, sputum microbiology and pulmonary function. Forty-one individuals (39 with R117H and two with R117C), 16 on an IVS8-5T background and 25 on an IVS8-7T background were identified. Twelve individuals presented clinically, four were siblings of known R117H/C compound heterozygotes and 25 were detected by newborn screening. Eleven of 14 of the IVS8-5T group (78%) with sweat chloride results available had sweat CI > 60 mmol x L(-1) compared to 5 (20%) of the R117H/7T group (Chi-squared=10.4, p=0.001). Two were pancreatic insufficient, both IVS8-5T. Two IVS8-5T individuals have recently died (aged 43 and 19) and of the 14 surviving IVS8-5T group, 11 (79%) are symptomatic compared to eight (32%) of the IVS8-7T individuals (Chi squared=6.1, p=0.01). In conclusion, most individuals with R117H/C on a IVS8-5T background have an elevated sweat chloride and clinical cystic fibrosis, which in some cases is severe. Most individuals with R117H/C on an IVS8-7T background do not have clinical cystic fibrosis but should be followed for the development of clinical disease. PMID- 11491165 TI - Increased nitrotyrosine in exhaled breath condensate in cystic fibrosis. AB - Exhaled nitric oxide (ENO), a marker of inflammation in airway diseases is decreased in cystic fibrosis (CF) patients, perhaps because nitric oxide (NO) is metabolized to oxidative end-products. A stable product, 3-nitrotyrosine, may indicate local formation of reactive nitrogen species. Whether NO metabolites in exhaled breath condensate may be increased in CF patients was investigated. The fractional concentration of ENO (Feno), nitrotyrosine and oxides of nitrogen in exhaled breath condensate from 36 stable CF patients were compared to 14 normal subjects using an enzyme immunoassay and fluorescence assay. Nitrotyrosine levels in breath condensate were increased significantly in stable CF patients, compared with normal subjects (25.3 +/- 1.5 versus 6.3 +/- 0.8 ng x mL(-1), p<0.0001). There was an inverse correlation between the levels of nitrotyrosine and the severity of lung disease. Feno levels were significantly lower in CF patients than in normal subjects (4.4 +/- 0.3 versus 5.6 +/- 0.4 (parts per billion), p<0.05). No correlation was found between nitrotyrosine and Feno levels in CF. There was no significant difference in the levels of nitrite and nitrate between CF patients and normals. The elevation in nitrotyrosine may reflect increased formation of reactive nitrogen species such as peroxynitrite or direct nitration by granulocyte peroxidases, indicating increased oxidative stress in airways of cystic fibrosis patients. PMID- 11491166 TI - Nasal potential difference measurements in patients with atypical cystic fibrosis. AB - The diagnosis of cystic fibrosis (CF) is based on characteristic clinical and laboratory findings. However, a subgroup of patients present with an atypical phenotype that comprises partial CF phenotype, borderline sweat tests and one or even no common cystic fibrosis transmembrane conductance regulator (CFTR) mutations. The aim of this study was to evaluate the role of nasal potential difference (PD) measurements in the diagnosis of CF patients with an atypical presentation and in a population of patients suspected to have CF. Nasal PD was measured in 162 patients from four different groups: patients with classical CF (n = 31), atypical phenotype (n = 11), controls (n = 50), and patients with questionable CF (n = 70). The parameter, or combination of nasal PD parameters was calculated in order to best discriminate all CF patients (including atypical CF) from the non-CF group. The patients with atypical CF disease had intermediate values of PD measurements between the CF and non-CF groups. The best discriminate model that assigned all atypical CF patients as CF used: e(response to chloride free and isoproterenol/response to amiloride) with a cut-off >0.70 to predict a CF diagnosis. When this model was applied to the group of 70 patients with questionable CF, 24 patients had abnormal PD similar to the atypical CF group. These patients had higher levels of sweat chloride concentration and increased rate of CFTR mutations. Nasal potential difference is useful in diagnosis of patients with atypical cystic fibrosis. Taking into account both the sodium and chloride transport elements of the potential difference allows for better differentiation between atypical cystic fibrosis and noncystic fibrosis patients. This calculation may assist in the diagnostic work-up of patients whose diagnosis is questionable. PMID- 11491167 TI - Lung cancer in patients with idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) was reported to be associated with increased risk of lung cancer as a result of the occurrence of atypical or dysplastic epithelial changes in fibrosis which progressed to invasive malignancy. In that situation, the cancer will develop in the area of major fibrosis. To investigate the direct relationship between fibrosis and cancer development, the real concordance rate of the two lesions in the chest computed tomography (CT) was analysed and compared to the histological types of lung cancer. The subjects included 63 patients with combined lung cancer and IPF (IPF-CA), 218 patients with lone IPF, and 2,660 patients with primary lung cancer. All patients were diagnosed at Asan Medical Center during the same period. The age, percentage of smokers, and the male sex were significantly higher in IPF-CA compared with lone IPF. The odds ratio of smoking was 2.71 compared to nonsmoking IPF controls. In IPF-CA, 56% of the cancer was located in the periphery of the lung and 52% in the upper lobe. The majority of the cancers (64%) were found in the nonfibrotic area at chest CT. The most frequent cell type was squamous cell carcinoma (35%), and there was no significant difference in the cancer cell type between IPF-CA and total lung cancer population. These findings suggest that in combined lung cancer and idiopathic pulmonary fibrosis patients, the features of the lung cancer are similar to the total lung cancer population. PMID- 11491168 TI - Type II alveolar epithelial cells and interstitial fibroblasts express connective tissue growth factor in IPF. AB - Connective tissue growth factor (CTGF) is a growth and chemotactic factor for fibroblasts encoded by an immediate early gene that is transcriptionally activated by transforming growth factor-beta. Previous studies have shown that both CTGF messenger ribonuclear acid (mRNA) and protein are expressed in renal fibrosis and bleomycin-induced pulmonary fibrosis in mice. The aim of the present study was to investigate the localization of CTGF protein and its mRNA expression in the fibrotic lung tissue of patients with idiopathic pulmonary fibrosis (IPF). Using human fibrotic lung tissue obtained from eight autopsy cases and four biopsy cases with IPF, immunohistochemical staining, in situ hybridization, and reverse transcription-polymerase chain reaction (RT-PCR) were performed. The cellular immunoreactivity for CTGF was markedly increased in the lung tissue of patients with IPF, compared to normal lungs. The immunolocalization of CTGF was confined predominantly to proliferating type II alveolar epithelial cells and activated fibroblasts. In the normal lung, type II alveolar epithelial cells stained for CTGF were sparsely distributed. CTGF mRNA was localized in proliferating type II alveolar epithelial cells and activated fibroblasts in the interstitium of fibrotic lung tissues. RT-PCR analysis showed that CTGF mRNA was expressed at a higher level in fibrotic lungs than in normal lungs. In both an autocrine and a paracrine manner, type II alveolar epithelial cells and activated fibroblasts may play a critical role in pulmonary fibrosis by producing connective tissue growth factor which modulates fibroblast proliferation and extracellular matrix production. PMID- 11491169 TI - Attenuation by oral N-acetylcysteine of bleomycin-induced lung injury in rats. AB - Antioxidant therapy may be useful in diseases with impaired oxidant-antioxidant balance such as pulmonary fibrosis. This study examines the effect of N acetylcysteine (NAC) on bleomycin-induced lung fibrosis in rats. NAC (3 mmol x kg(-1); oral) was given daily from 1 week prior to a single intratracheal instillation of bleomycin (2.5 U x kg(-1)) or saline, until 14 days postinstillation. NAC partially decreased the augmented collagen deposition in bleomycin-exposed rats (hydroxyproline content was 4,354+/-386 and 3,416+/-326 microg x lung(-1) in vehicle-treated and NAC-treated rats, respectively; p < 0.05). The histological assessment using a semiquantitative score showed less collagen deposition and inflammatory cells in NAC-treated rats compared to those receiving bleomycin alone. NAC failed to inhibit the bleomycin-induced increases in lung wet weight and in cell counts and protein levels of bronchoalveolar lavage fluid, but significantly increased total glutathione and taurine levels in bronchoalveolar lavage fluid. These results indicate that oral N-acetylcysteine improves the pulmonary antioxidant protection and may be useful in reducing lung damage produced by bleomycin. PMID- 11491170 TI - ABO/Secretor genetic complex and susceptibility to asthma in childhood. AB - A positive association has recently been reported in adult subjects between O/nonSecretor phenotype and asthma. To confirm this association, this study investigated the joint ABO/Secretor phenotype in a cohort of 165 asthmatic children. Three-hundred and sixty-two consecutive newborn infants from the same population were also studied as controls. The proportion of O/nonSecretor in asthmatic children was higher than in controls, thus confirming the association found in adults. The association was more marked in males than in females. In males, the pattern of association between the joint ABO/Secretor phenotype and asthma is dependent on the age at on-set of symptoms. Since the oligosaccharide composition of cell membrane and mucosal secretions is controlled by the cooperative interaction of ABO and Secretor genes, and since such composition influences the adhesion of infectious agents, the age pattern could reflect a more general interaction between developmental maturation and oligosaccharide structure concerning their effects on susceptibility to viral and bacterial agents. PMID- 11491171 TI - Effect of airway opening manoeuvres on thoraco-abdominal asynchrony in anaesthetized children. AB - Thoraco-abdominal asynchrony is frequently encountered during inhalation anaesthesia in children with adenotonsillar hypertrophy causing an upper airway obstruction. The study goal was to evaluate the impact of different airway opening manoeuvres on thoraco-abdominal asynchrony as a measure of airway obstruction. Thirty anaesthetized children (aged 2-8 yrs; sevoflurane 3% in 50% oxygen/nitrous oxide) were studied prior to elective adenotonsillectomy using respiratory inductance plethysmography to record ribeage and abdominal wave forms as a basis for calculation of the phase angle. Five airway situations were compared: 1) baseline (unsupported mandible); 2) chin lift; 3) chin lift combined with continuous positive airway pressure of 10 cmH2O; 4) jaw thrust; and 5) jaw thrust combined with continuous positive airway pressure of 10 cmH2O. Three children had complete upper airway obstruction at baseline and were excluded from the study. With chin lift, thoraco-abdominal asynchrony improved in three patients, worsened in three patients and was unchanged in 21 patients. Additional continuous positive airway pressure during chin lift did not markedly reduce thoraco-abdominal asynchrony (phase angle 89 +/- 43 , p = 0.33). Jaw thrust resulted in a significant decrease of the phase angle (from 106 +/- 53 at baseline to 65 +/- 49 , p < 0.01); when combined with continuous positive airway pressure, no further effect on thoraco-abdominal asynchrony was found (72 +/- 44). In anaesthetized children with adenotonsillar hypertrophy, airway opening manoeuvres have distinct effects on thoraco-abdominal asynchrony. Delivery of continuous positive airway pressure and jaw thrust can be the first airway opening manoeuvres to improve breathing patterns. Chin lift without additional continuous positive airway pressure should be used with caution in these patients because it may convert partial into almost complete airway obstruction. PMID- 11491172 TI - How should airways resistance be measured in young children: mask or mouthpiece? AB - The reproducibility and acceptability of airways resistance measurements using the interrupter technique (MicroRint) obtained using a mouthpiece were compared with those using a face mask. Fifty children aged 4-7 yrs performed four sets of six Rint measurements; two using a mouthpiece and two using a face mask with integral mouthpiece. Complete data were obtained from 45 (90%) children using the mouthpiece and 43 (86%) children using the mask. The two methods were equally repeatable with comparable intraclass correlation coefficients (ICC) and coefficients of variation. Mean Rint values obtained using the mouthpiece were significantly lower than those using the face mask ((mean+/-SD) mouthpiece=0.81+/ 0.18 kPa x L(-1) x s, mask=0.88+/-0.24 kPa x L(-1) x s p=0.0002). Although the mean paired differences between the two methods were small (0.07 kPa x L(-1) x s), the ICC and limits of agreement confirmed that the two methods could not be used interchangeably. Sixty-seven per cent of children preferred the face mask but this was more time-consuming (p = 0.03). Children did not produce more repeatable results using their preferred method, nor did they improve with practice. Repeatable airway resistance measurements using the interrupter technique can be obtained from young children using either a mouthpiece or a face mask, but there are significant clinical and statistical differences between the results obtained. PMID- 11491173 TI - Noninvasive pressure preset ventilation for the treatment of Cheyne-Stokes respiration during sleep. AB - Cheyne-Stokes respiration (CSR) during sleep is common in patients with congestive heart failure (CHF). This pattern of breathing fragments sleep, leading to daytime symptoms of sleepiness and fatigue. It was hypothesized that by controlling CSR with noninvasive pressure preset ventilation (NPPV), there would be a decrease in sleep fragmentation and an improvement in sleep quality. Nine patients (eight males, one female; mean +/- SD 65 +/- 11 yrs) with symptomatic CSR diagnosed on overnight polysomnography (apnoea/hypopnoea index (AHI) 49 +/- 10 x h(-1), minimum arterial oxygen saturation (Sa,O2, 77 +/- 7%) and CHF (left ventricular ejection fraction 25 +/- 8%) were studied. After a period of acclimatization to NPPV (variable positive airway pressure (VPAP) II ST, Sydney, NSW, Australia and bilevel positive airway pressure (BiPAP), Murraysville, PA, USA), sleep studies were repeated on therapy. NPPV almost completely abolished CSR in all patients with a reduction in AHI from 49 +/- 10 to 6 +/- 5 x h(-1) (p<0.001). Residual respiratory events were primarily due to upper airway obstruction at sleep on-set. Arousal index was markedly decreased from 42 +/- 6 to 17 +/- 7 x h(-1) (p <0.001). Sleep architecture showed a trend toward improvement with a reduction in stage 1 and 2 (79 +/- 7% during the diagnostic night versus 72 +/- 10% during NPPV, (p=0.057)), whilst sleep efficiency, slow-wave sleep (SWS), and rapid eye movement (REM) were not altered. Controlling Cheyne-Stokes respiration with noninvasive pressure preset ventilation resulted in reduced arousal and improved sleep quality in the patients with congestive heart failure. Noninvasive pressure preset ventilation should be considered a potential therapy for Cheyne-Stokes respiration in congestive heart failure in those patients who do not respond or fail to tolerate nasal continuous positive airway pressure therapy. PMID- 11491174 TI - The sleep apnoea/hypopnoea syndrome depresses waking vagal tone independent of sympathetic activation. AB - The modest daytime hypertension and sympathetic upregulation associated with the sleep apnoea/hypopnoea syndrome (SAHS), does not explain the relatively large increased risk of cardiac morbidity and mortality in the SAHS patients population. Therefore, efferent vagal and sympathetic activity was evaluated during wakefulness in SAHS subjects and matched healthy controls, in order to determine if vagal downregulation may play a role in the aetiology of cardiac disease in the SAHS. The awake autonomic nervous system function of 15 male subjects, with mild-to-moderate SAHS was compared to that of 14 healthy controls matched for age, body mass index, gender and blood pressure. All subjects were free from comorbidity. Vagal activity was estimated from measurements of heart rate variability high frequency power (HF) and sympathetic activity was measured from urine catecholamine excretion. The %HF power was significantly (p < 0.03) reduced in SAHS patients (10+/-1.6 (mean+/-SEM)) as compared to controls (17 +/- 3). In addition, HF power correlated with the apnoea/hypopnoea index in the SAHS subjects (R = -0.592, p = 0.02). There was no statistically significant difference in the daytime excretion of nonadrenaline between control (242 +/- 30 nmol x collection(-1)) and SAHS (316 +/- 46 nmol x collection(-1)) subjects (p = 0.38). In these sleep apnoea/hypopnoea syndrome patients there was limited evidence of increased waking levels of urine catecholamines. The principal component altering waking autonomic nervous system function, in the sleep apnoea/hypopnoea syndrome subjects, was a reduced daytime efferent vagal tone. PMID- 11491175 TI - Glucocorticoid treatment reduces exhaled nitric oxide in cystic fibrosis patients. AB - In cystic fibrosis (CF), low concentrations of exhaled nitric oxide (NO) and reduced expression of inducible nitric oxide synthase (iNOS) in airway epithelium have been reported. However, abundant iNOS expression has been found in the subepithelial tissues and elevated concentrations of NO metabolites in breath condensate and sputum. These conflicting results may be explained by increased scavenging of NO by superoxide radicals, resulting in rapid conversion to peroxynitrite, so that only a small proportion of the NO produced in the lung tissue reaches the airway lumen. If iNOS were active in the CF lung, exhaled NO would be further reduced by glucocorticoid treatment. CF patients (n = 13) were recruited to a double-blind, placebo-controlled study with crossover. Treatment comprised prednisolone or placebo for 5 days with a 9 day washout. After each treatment, exhaled NO was measured, spirometry performed and blood collected for measurement of serum nitrogen dioxide/nitrous oxide (NO2/NO3). Ten patients (8 male) completed the study. Following prednisolone treatment (mean +/- SD) exhaled NO concentration (3.1 +/- 1.6 parts per billion (ppb)) was significantly reduced versus placebo treatment (4.9 +/- 4.2 ppb; p<0.05, Wilcoxon signed-rank test). Spirometric indices and serum NO2/NO3 concentration were unchanged. These findings support the hypothesis that glucocorticoids suppress nitric oxide production in cystic fibrosis airways by reducing inducible nitric oxide synthase expression or by inhibiting recruitment of neutrophils, cells which express inducible nitric oxide synthase. PMID- 11491176 TI - Noninvasive ventilation and obstructive lung diseases. AB - The key role of noninvasive positive pressure ventilation (NPPV) is well documented in chronic obstructive pulmonary disease (COPD) patients with acute respiratory failure (ARF) since it may avoid endotrachal intubation in >50% of cases when used as the initial treatment. However, currently only minimal data is available to assess usefulness of NPPV in COPD patients on a long-term basis. Even if such studies are difficult to manage, there is clearly a need for prospective studies comparing long-term oxygen therapy (LTOT) and NPPV in the most severe COPD in a large amount of patients and on a real long-term basis of several years. Two randomized prospective studies are being completed in Europe and the first preliminary results show that NPPV is associated with a reduction of hospitalization for chronic respiratory failure decompensation. The main beneficial effect of long-term mechanical ventilation in COPD patients with chronic respiratory failure implies a correction of nocturnal hypoventilation that could persist beyond the ventilation period because of a temporary improvement in carbon dioxide sensitivity that is often blunted in these patients. A synthesis from the literature suggest to consider NPPV for severe COPD patients who present with chronic hypoxia and hypercapnia and develop an unstable respiratory condition. Instability may be appreciated on a clinical basis and confirmed by a progressive worsening of arterial blood gas tensions, leading to frequent cardiorespiratory decompensations with ominous ARF episodes. NPPV should also be considered after an ARF episode successfully treated by noninvasive ventilation but with the impossibility to wean the patient from the ventilator. Thus, noninvasive positive pressure ventilation could be proposed as a preventive treatment in severe chronic obstructive pulmonary disease patients with unstable respiratory condition associated with fluctuating hypercapnia before, during and after an acute respiratory failure episode, avoiding the need for a tracheotomy. Adjunction of noninvasive ventilation to exercise rehabilitation is under evaluation. PMID- 11491177 TI - Influenza: vaccination and treatment. AB - Few conditions exert such an enormous toll of absenteeism, suffering, medical consultations, hospitalization, death and economic loss as influenza. Patients at high risk of complications and mortality include the elderly and those with pre existing cardiopulmonary disease. The outbreak in 1997 in Hong Kong, of avian H5N1 influenza in man, which resulted in six deaths among 18 hospitalized cases, and the recent isolation of H9N2 viruses from two children in Hong Kong, are reminders that preparation must be made for the next pandemic. Since the 1970s, efforts to control influenza have mostly focussed on the split product and surface antigen vaccines. These vaccines are of proven efficacy in healthy adults and are effective in elderly people with and without medical conditions putting them at high risk of complications and death following influenza infection. However, vaccine coverage is patchy and often low, and outbreaks of influenza are not uncommon in well-immunized residents of nursing homes. New vaccines and methods of vaccine delivery are being developed in attempts to overcome the limitations of existing vaccines. The antiviral drugs amantadine and rimantadine were developed in the 1960s, but have not been used widely due to their spectrum of activity, rapid emergence of resistance, and adverse effects associated with amantadine. The site of enzyme activity of the influenza neuraminidase is highly conserved between types, subtypes and strains of influenza and has emerged as the target of an exciting new class of antiviral agents that are effective both prophylactically and as therapy. PMID- 11491178 TI - Small airways diseases: detection and insights with computed tomography. AB - Diseases affecting the small airways are difficult to detect by traditional diagnostic tests. Widespread involvement is needed before symptoms and abnormalities on pulmonary function testing or chest radiography become apparent. Obstruction of the bronchioles may be detected indirectly by computed tomography (CT) because regional under-ventilation results in reduced perfusion which in turn is shown as a mosaic attenuation pattern of the lung parenchyma. When there is inflammation of the bronchioles with accompanying exudate, the airways may become directly visible on CT, for example in cases of diffuse panbronchiolitis. Quantification of the various morphological features of small airways disease is possible from CT images and this increased precision has aided investigations of structure/function relationships. An understanding of the pathology and microscopic distribution of disease in relation to the airways allows some prediction of the likely computed tomography appearances in this wide spectrum of conditions, and thus helps to refine the differential diagnosis. PMID- 11491179 TI - Pharmacological treatment of the biochemical defect in cystic fibrosis airways. AB - The understanding of the biochemical defect in cystic fibrosis (CF) has advanced considerably since discovery of the CF gene in 1989 and characterization of its product. Studies showing that the abnormality in chloride flux could be corrected by transfection of wild-type cystic fibrosis transmembrane conductance regulator (CFTR) complimentary deoxyribonucleic acid (cDNA) have led to gene therapy trials on both sides of the Atlantic. However, gene therapy as a treatment for CF has yet to be realized. Pharmacological manipulation of the biochemical defect may provide an alternative or complementary approach to treatment. This review will discuss pharmacological agents in development which could correct the abnormal ion movement. The mechanisms of action of these pharmacological agents can be divided broadly into drugs which affect the most common CF mutation, deltaF508, which increase trafficking of the mutant CF protein to the apical membrane; drugs which increase chloride secretion; and drugs which reduce sodium reabsorption across the apical membrane. Treatment options for cystic fibrosis have developed rapidly since discovery of the cystic fibrosis gene over a decade ago. The targeting of specific therapies for particular cystic fibrosis genotypes and the use of combination treatments of chloride channel openers with sodium channel blockers are likely to be key advances in the next decade. PMID- 11491180 TI - A small outbreak of Legionnaires' disease in a cargo ship under repair. AB - It was reported that two mechanics working on a cargo ship under repair in the port of Barcelona had died after having fever. An investigation was made into the possibility of any additional cases and the presence of Legionella pneumophila in the ship they were repairing and in their hotel. The contaminated water system was treated with sodium hypochlorite. Both patients died after having been repeatedly diagnosed as having influenza. The two cases occurred among those who had been working with the pump of the ship's water system, while no cases were observed among the other workers (p = 0.02). Various serogroups of L. pneumophila were isolated from the ship's water pump and distribution system. However, organism of serogroup 1, subgroup Pontiac (Knoxville) were identified with identical deoxyribonucleic acid (DNA) patterns in the lung tissue of one patient and in the cooling water circuit valve of the ship's water pump. The first postintervention control water samples showed no further growth of legionella, but serogroups 4 and 8 were identified 8 months later. This legionellosis outbreak, although small, was highly lethal, probably due to the high levels of bacteria to which the patients were exposed and also because of the failure of correct diagnosis. International recommendations on prevention and control of legionellosis, which include ships under repair, are required. PMID- 11491181 TI - A 48-yr-old female with headache and dyspnoea. Pulmonary arteriovenous malformation. PMID- 11491182 TI - Comparative efficacy and safety of mometasone furoate dry powder inhaler and budesonide Turbuhaler. PMID- 11491183 TI - Lung cancer in young females. PMID- 11491184 TI - Long-term treatment of pulmonary hypertension with aerosolized iloprost. PMID- 11491185 TI - Risk of myocardial ischaemia and beta-adrenoceptor agonists. AB - Modern therapy for both cardiovascular disease and obstructive lung disease involves diametrically opposed manipulations of the beta-adrenoceptor. Beta agonists reduce airflow limitation and improve symptoms among patients with obstructive lung disease while beta-blockers reduce symptoms, recurrent myocardial ischaemia and all-cause mortality among patients with ischaemic heart disease. There is biological plausibility for beta-agonists leading to adverse cardiovascular outcomes, and observational trials have raised concern about the safety of beta-agonists among patients with cardiovascular disease. Although there are many potential causal and noncausal explanations for these observational findings, the implications from these studies are the same. Physicians should be careful when prescribing beta-agonists for patients at risk for ischaemic heart disease. Furthermore, careful consideration should be given to distinguish symptoms caused by cardiovascular versus respiratory aetiologies. PMID- 11491186 TI - Oncolytic viruses as therapeutic agents. AB - The concept of using viruses as oncolytic agents has a long history. However, relatively new developments are the use of these viruses as gene delivery vehicles and the restriction of viral replication and lysis to tumour cells. The latter is attempted by the use of tumour-specific promoters, which transcriptionally target viral genes involved in replication, or by deletion of viral functions dispensable for replication in tumour cells but essential for productive infection of normal cells. In addition, retargeting of the viral tropism towards tumours by capsid modifications has been examined. Although much progress has been made in developing oncolytic vectors for clinical use, there is still a long way to go to determine which combinations of virus, gene therapy, surgery, radiation, and/or chemotherapy will provide improved therapy for the control and eradication of a variety of human cancers. First controlled clinical trials with an oncolytic adenovirus in combination with chemotherapy have shown encouraging antineoplastic activity. For future vector developments it will be crucial to achieve maximum vector distribution and transgene expression within tumours, to trigger a specific systemic immune effector response against treated and untreated lesions, and to modulate the immune system to avoid immune-mediated inactivation or destruction of the virus. In the context of replication-competent vectors, suicide genes might be used as fail-safe mechanism in the case of a runaway infection. PMID- 11491187 TI - Assessing the role of oestrogen in the prevention of cardiovascular disease. AB - Coronary heart disease remains the leading cause of death in postmenopausal women around the world, and interest is high in discovering the best treatments and methods of prevention for this disease. For many years, it appeared that one such treatment could be oestrogen, because of its beneficial effects on the vascular endothelium and on cholesterol concentrations. However, recent clinical trials have shown no beneficial effect of long-term hormone replacement therapy (HRT) on risk for major cardiovascular events among women with established coronary disease. These surprising findings have led to still further analyses to elucidate plausible explanations. This paper will review the results from recent trials and clinical studies of HRT, as well as ongoing trials that continue to examine the role of oestrogen in the treatment and prevention of cardiovascular disease. PMID- 11491188 TI - The transcription factor Egr-1: a potential drug in wound healing and tissue repair. AB - In the United States, between 40 and 90 million hospital days are lost per year as a result of trauma and surgical procedures which result in the loss of functional tissue. This is estimated to cost the economy and healthcare providers in excess of US$ 500 billion, a figure that is increasing because of extending population lifespan. Tissue engineering and gene therapies are radical new treatments that are aimed at tissue regeneration ranging from dermal, osteal and occular repair to the replacement of failing tissue with entire biosynthetic organs. Over the last decade, numerous proteins have been identified that are able to direct the synthesis of new tissue. Such proteins include growth factors, cytokines and, more recently, transcription factors. PMID- 11491189 TI - Be generic and specific: quality of life measurement in clinical studies. PMID- 11491190 TI - Measuring quality of life in evaluating clinical interventions: an overview. AB - Health professionals provide care with some purpose in mind. It is reasonable to assume that they are therefore interested in observing and measuring health outcomes. In the recent past, it has become fashionable to describe health outcomes in terms of their impact on 'quality of life', a widely used term that is usually left undefined. Measures of health-related quality of life (HRQoL) are now in common usage in clinical studies. The construction of both generic and condition-specific measures of HRQoL relies heavily on the choice of descriptive domains, a process that often reflects the personal values of the developer of the instrument. Subjective valuation is a recurring phenomenon in the construction and application of HRQoL measures. These values should not be restricted to the private judgement of the clinician alone. The challenge for those evaluating clinical interventions is to make such values explicit and to extend the franchise to society as a whole. PMID- 11491191 TI - The 15D instrument of health-related quality of life: properties and applications. AB - The 15D is a generic, comprehensive, 15-dimensional, standardized, self administered measure of health-related quality of life (HRQoL) that can be used both as a profile and single index score measure. This paper examines the acceptability, reliability, validity, discriminatory power and responsiveness to change of its health state descriptive system and valuation system and presents some examples of applications. As a profile measure on roughly comparable dimensions the 15D performs equally well as the Nottingham Health Profile (NHP) and SF-20, in some respects even better, and clearly better than EQ-5D. The remaining nine to ten dimensions of the 15D provide a large reserve in terms of discriminatory power and responsiveness to change. The valuation system is based on an application of the multiattribute utility theory. The single index score (15D score) on a 0-1 scale, representing the overall HRQoL, is calculated from the health state descriptive system by using a set of population-based preference or utility weights. The 15D scores are shown to be highly reliable, sensitive and responsive to change, generalisable at least in Western-type societies, and particularly valid for deriving quality-adjusted life years (QALYs) gained for resource allocation purposes. The instrument is recommended by the Washington Panel and is available in several languages for clinical economic evaluation and population studies. PMID- 11491192 TI - EQ-5D: a measure of health status from the EuroQol Group. AB - Established in 1987, the EuroQol Group initially comprised a network of international, multilingual and multidisciplinary researchers from seven centres in Finland, the Netherlands, Norway, Sweden and the UK. Nowadays, the Group comprises researchers from Canada, Denmark, Germany, Greece, Japan, New Zealand, Slovenia, Spain, the USA and Zimbabwe. The process of shared development and local experimentation resulted in EQ-5D, a generic measure of health status that provides a simple descriptive profile and a single index value that can be used in the clinical and economic evaluation of health care and in population health surveys. Currently, EQ-5D is being widely used in different countries by clinical researchers in a variety of clinical areas. EQ-5D is also being used by eight out of the first 10 of the top 50 pharmaceutical companies listed in the annual report of Pharma Business (November/December 1999). Furthermore, EQ-5D is one of the handful of measures recommended for use in cost-effectiveness analyses by the Washington Panel on Cost Effectiveness in Health and Medicine. EQ-5D has now been translated into most major languages with the EuroQol Group closely monitoring the process. PMID- 11491193 TI - Introducing economic and quality of life measurements into clinical studies. AB - Although the collection of cost and quality of life data alongside clinical studies generates detailed patient level data in a timely fashion, it also raises practical and methodological challenges. These include the fact that the settings and patients enrolled in trials may not be typical of those found in regular clinical practice, that costs and quality of life may be influenced by the trial protocol, that the clinical alternatives compared in trials may not be the most relevant for cost-effectiveness assessments, that the length of follow-up may be too short to observe changes in cost and quality of life, and that adding these data will increase the overall measurement burden in the trial. This paper discusses these challenges and the ways in which they might be overcome, focussing particularly on preference-based measures of quality of life. In particular, recommendations are given for choosing the range of quality of life instruments, sample size calculations for quality of life measurement and the measurement of quality of life in multinational studies. PMID- 11491194 TI - The RAND-36 measure of health-related quality of life. AB - The RAND-36 is perhaps the most widely used health-related quality of life (HRQoL) survey instrument in the world today. It is comprised of 36 items that assess eight health concepts: physical functioning, role limitations caused by physical health problems, role limitations caused by emotional problems, social functioning, emotional well-being, energy/fatigue, pain, and general health perceptions. Physical and mental health summary scores are also derived from the eight RAND-36 scales. This paper provides example applications of the RAND-36 cross-sectionally and longitudinally, provides information on what a clinically important difference is for the RAND-36 scales, and provides guidance for summarizing the RAND-36 in a single number. The paper also discusses the availability of the RAND-36 in multiple languages and summarizes changes that are incorporated in the latest version of the survey. PMID- 11491195 TI - A comparison of the Assessment of Quality of Life (AQoL) with four other generic utility instruments. AB - As part of the validation of the Assessment of Quality of Life (AQoL) instrument comparisons were made between five multiattribute utility (MAU) instruments, each purporting to measure health-related quality of life (HRQoL). These were the AQoL, the Canadian Health Utilities Index (HUI) 3, the Finnish 15D, the EQ-5D (formerly the EuroQoL) and the SF6D (derived from the SF-36). The paper compares absolute utility scores, instrument sensitivity, and incremental differences in measured utility between different instruments predicted by different individuals. The AQoL predicted utilities are similar to those from the HUI3 and EQ-5D. By contrast the 15D and SF6D predict systematically higher utilities, and the differences between individuals are significantly smaller. There is some evidence that the AQoL has greater sensitivity to health states than other instruments. It is concluded that at present no single MAU instrument can claim to be the 'gold standard', and that researchers should select an instrument sensitive to the health states they are investigating. Caution should be exercised in treating any of the instrument scores as representing a trade-off between length of life and HRQoL. PMID- 11491196 TI - Health state values from multiattribute utility instruments need correction. AB - Cost-utility analysis uses the quality-adjusted life year (QALY) as a measure of the benefit of health interventions. It presupposes the assignment of utility scores to different states of health on a scale from zero (dead) to unity (healthy). A number of so-called multiattribute utility (MAU) instruments are available for this purpose. Analysts who wish to use MAU instruments in economic evaluations of health programmes and technologies may improve their performance by conducting two different analyses: the first is a conventional cost-utility study, in which the utilities from MAU instruments are used as they stand, and the second is a study in which the utilities are transformed into numbers that also encapsulate concerns for giving priority to the worst off. The term 'cost value analysis' is used for the latter, broader approach. A figure is offered as a preliminary tool to help conduct the required transformations. PMID- 11491197 TI - The Health Utilities Index (HUI) system for assessing health-related quality of life in clinical studies. AB - This paper reviews the Health Utilities Index (HUI) systems as means to describe health status and obtain utility scores reflecting health-related quality of life (HRQoL). The HUI Mark 2 (HUI2) and Mark 3 (HUI3) classification and scoring systems are described. The methods used to estimate multiattribute utility functions for HUI2 and HUI3 are reviewed. The use of HUI in clinical studies for a wide variety of conditions in a large number of countries is illustrated. HUI provides a comprehensive description of the health status of subjects in clinical studies. HUI has been shown to be a reliable, responsive and valid measure in a wide variety of clinical studies. Utility scores provide an overall assessment of the HRQoL of patients. Utility scores are also useful in cost-utility analyses and related studies. General population norm data are available. The widespread use of HUI facilitates the interpretation of results and permits comparisons. HUI is a useful tool for assessing health status and HRQoL in clinical studies. PMID- 11491198 TI - The role of troponin abnormalities as a cause for stunned myocardium. AB - Myocardial stunning is a form of ischemic injury, which occurs with transient ischemia followed by re-establishment of flow, and which results in reversible cardiac dysfunction. There is evidence that the molecular defect in stunning is at the level of the contractile apparatus. Selective proteolysis of the myofilament protein, troponin I, appears to underlie the phenotype of stunning in some models, but other myofilament protein modifications may also have a role. PMID- 11491199 TI - Evidence for stunned myocardium in humans: a 2001 update. AB - This article describes clinical situations in which stunning occurs and updates previous reviews on the topic. Stunning following angioplasty, angina and exercise-induced ischemia, infarction, and after cardiac surgery are described. In addition, newer concepts regarding stunning, including neurogenic stunned myocardium, are discussed. Left atrial stunning following cardioversion is a recently recognized phenomenon with important clinical implications, but differs from the original concept of post-ischemic stunning. PMID- 11491200 TI - Clinical manifestations of myocardial stunning. AB - The phenomenon of myocardial stunning has been observed in all animal species studied. The possible occurrence of myocardial stunning in man has been demonstrated after either regional ischemia (such as exercise-induced angina, vasospastic or unstable angina) or after global ischemia (i.e., after cardioplegic arrest during cardiac surgery, or cardiac arrest, or heart transplantation). Finally, it may also be observed in patients with acute myocardial infarction, subjected to recanalization therapy, because viable myocardium, salvaged by reperfusion, may remain stunned, with delayed contractile recovery. Occurrence of stunning may aggravate hemodynamic conditions in already unstable patients, and it may lead to underestimation of the extent of myocardium salvaged by thrombolysis. Repeated episodes of stunning may lead to a condition of apparently 'chronic' contractile dysfunction that may be difficult to differentiate from hibernation, because of the technical difficulties in accurately measuring myocardial blood flow in patients, and because both phenomena may coexist and overlap in the same patient. In addition, recent evidence suggests that repeated episodes of stunning may lead to a progressive worsening of the residual contractile dysfunction and to longer recovery times, and it has thus been suggested, and it is much debated, that hibernation might at least in part be the consequence of repetitive episodes of stunning. PMID- 11491201 TI - Pharmacologic treatment of the stunned myocardium: the concepts and the challenges. AB - During the past two decades (i.e., since 1980), in excess of 1000 published papers have focused on the phenomenon of the 'stunned myocardium', with many of these studies seeking to identify mechanisms-based treatment strategies to attenuate post-ischemic contractile dysfunction. Early investigations focused largely on abrogating the deleterious effects of oxygen-derived free radicals and unfavorable alterations in calcium homeostasis, both considered to contribute significantly to the pathogenesis of the stunned myocardium. More recently, favorable results have also been obtained using a somewhat different paradigm: that is, attempting to capitalize on endogenous cardioprotective mediators, most notably adenosine, nitric oxide, and the ATP-sensitive potassium channel. Now that potential therapeutic candidates have been identified in the experimental laboratory, the as-yet unmet challenge is to translate this information into the design of effective pharmacologic therapies to treat myocardial stunning in the clinical arena. PMID- 11491202 TI - Lessons from experimental models of hibernating myocardium. AB - Chronic animal models of viable dysfunctional myocardium are now available that recapitulate most if not all of the physiological findings in humans with hibernating myocardium. These include chronic reductions in resting perfusion and contractile function, critical limitations in coronary flow reserve and increased uptake of 18F-2-deoxyglucose. These changes occur in the absence of infarction or necrosis and are accompanied by regional reductions in sarcoplasmic reticulum calcium-handling proteins and myocyte loss that arise secondary to apoptosis. Longitudinal studies of viable dysfunctional myocardium indicate that a state of chronic stunning with normal resting flow precedes the development of hibernating myocardium but these are distinct entities within a continuum of chronic adaptations to ischemia. This indicates that reductions in resting flow are the result rather than cause of chronic contractile dysfunction. Thus, the original concept proposing an acute prolonged reduction in flow as the initial stimulus producing hibernating myocardium needs to be revised. PMID- 11491203 TI - Clinical pathophysiology of hibernating myocardium. AB - Our current knowledge of the pathophysiology of chronic hibernating myocardium is mainly based on results from clinical studies, because of the absence of appropriate and validated animal models. These clinical observations have given rise to two major controversies: the role of reduced blood flow and that of histological changes in the hibernating segments. In this review, these two subjects will be briefly discussed, and put into the perspective of findings emerging from recently developed animal models. PMID- 11491204 TI - Clinical importance of stunned and hibernating myocardium. AB - Myocardial stunning and hibernation are states of potentially reversible myocardial dysfunction, which were first described more than 20 years ago (c.1980). Important advances have now been made in the ability to detect stunned and hibernating myocardium, as well as in the understanding of the impact of these conditions on patient outcomes. We discuss here the clinical importance of stunned and hibernating myocardium for patients with several common cardiac conditions. PMID- 11491205 TI - Determinants of left ventricular systolic function after acute myocardial infarction: the role of residual myocardial ischaemia. AB - BACKGROUND: Left ventricular systolic function (LVSF) is one of the major determinants of survival after acute myocardial infarction (AMI). Some factors such as the infarct size and localization, and the patency of the infarct-related artery are known determinants of LVSF. However, the long-term effect of myocardial ischaemia on LVSF has been poorly studied in clinical settings. OBJECTIVES: To assess the acute and long-term effects of myocardial ischaemia on LVSF in patients recovering from an AMI. METHODS: A cohort of 74 patients recovering from AMI was studied. Myocardial ischaemia was detected by means of ambulatory electrocardiogram (ECG) monitoring at recruitment (4+/-2 days after AMI), exercise ECG test and stress echocardiography at discharge (7+/-4 days after AMI). LVSF was studied by means of two-dimensional echocardiography at recruitment, at discharge, and at 1, 3, 6 and 12 months after AMI. RESULTS: Patients with myocardial ischaemia on ambulatory ECG monitoring and stress echocardiography had worse LVSF at recruitment than those without ischaemia. The presence of myocardial ischaemia on ambulatory ECG monitoring was an independent determinant of LVSF at recruitment together with infarct localization and size (assessed by creatine kinase MB isoenzyme (CK-MB) levels). Patients with signs of myocardial ischaemia on ambulatory ECG monitoring and stress echocardiography had a progressive left ventricular dysfunction compared with those without ischaemia. CONCLUSIONS: Residual ischaemia is an independent determinant of LVSF after AMI and its presence implied a progressive worsening of the LVSF. Because left ventricular systolic dysfunction is a major determinant of survival after AMI, its precursors, among them residual myocardial ischaemia, should be identified. Treatment of ischaemia is known to be associated with improved prognosis and improved LVSF. PMID- 11491206 TI - Leukocyte attack in a 3D human coronary in-vitro model. AB - OBJECTIVE: The importance of peripheral blood leukocytes for the development of early atherosclerosis and restenosis has confronted cardiologists with classical hematologic issues. Three-dimensional human coronary in-vitro units of leukocyte attack (3DLA-units) open the field for exact studies of leukocyte attack and its subsequent effects on human medial coronary smooth muscle cells (HCMSMC). METHODS: Central part of 3DLA-units are polycarbonate membranes with a pore size of 5 microm that correspond to the internal elastic membrane. Human coronary endothelial cells (HCAEC) were cultured on one side of the membranes, HCMSMC on the other side. Before leukocyte attack expression of adhesion molecules was up regulated by tumour necrosis factor-alpha (TNF-alpha). Leukocyte attack was mimicked by selective adding of human monocytes (MC), respectively human CD4+ lymphocytes (CD4+-LC) to the HCAEC side of the 3DLA-units. Three-dimensional leukocyte attack units were fixed and stained after a period of 30 min, 1, 2, 3, 4, 6, and 24 h. Cell divisions of HCMSMC were analysed by measuring the uptake of bromodeoxyuridine (BrdU). RESULTS: Monocytes were able to adhere to the endothelial surface, pass through the filter-pores, and penetrate the HCMSMC side of the 3DLA-units. Human CD4+-lymphocytes (CD4+-LC) only attached to the HCAEC side, and no chemotaxis to the HCMSMC side was detected. Proliferation of HCMSMC was increased 2.9-fold (P< 0.001) after selective MC-attack and 3.5-fold after selective MC-attack and TNF-alpha stimulus. No significant increase was found after selective CD4+-LC attack, a significant increase (2.1-fold; P < 0.001) was seen after selective CD4+-LC attack and TNF-alpha, stimulus. CONCLUSIONS: Within the given limitations of the model the study emphasizes a predominance of MC in comparison to CD4+-LC in the process of adhesion, chemotaxis, and triggered reactive proliferation of co-cultured HCMSMC within the first 24 h after leukocyte attack. 3DLA-units offer an elegant method to study directly the effects of intravascular and intramural treatment strategies. PMID- 11491207 TI - The impact of pioglitazone on glycemic control and atherogenic dyslipidemia in patients with type 2 diabetes mellitus. AB - BACKGROUND: To evaluate the glycemic control, lipid effects, and safety of pioglitazone in patients with type 2 diabetes mellitus. DESIGN AND METHODS: Patients (n = 197) with type 2 diabetes mellitus, a hemoglobin A1c (HbA1c) > or = 8.0%, fasting plasma glucose (FPG) > 7.7 mmol/l (140 mg/dl), and C-peptide > 0.331 nmol/l (1 ng/ml) were enrolled in this 23-week multi-center (27 sites), double-blind clinical trial and randomized to receive either a placebo or pioglitazone HCl 30 mg (pioglitazone), administered once daily, as monotherapy. Patients were required to discontinue all anti-diabetic medications 6 weeks before receiving study treatment. Efficacy parameters included HbA1c fasting plasma glucose (FPG), serum C-peptide, insulin, triglycerides (Tg), and cholesterol (total cholesterol [TC], high-density lipoprotein-cholesterol [HDL C], low-density lipoprotein-cholesterol [LDL-C]). Adverse event rates, serum chemistry, and physical examinations were recorded. RESULTS: Compared with placebo, pioglitazone significantly (P= 0.0001) reduced HbA1c (-1.37% points), FPG (-3.19 mmol/l; -57.5 mg/dl), fasting C-peptide (-0.076+/-0.022 nmol/l), and fasting insulin (-11.88+/-4.70 pmol/l). Pioglitazone significantly (P < 0.001) decreased insulin resistance (HOMA-IR; -12.4+/-7.46%) and improved beta-cell function (Homeostasis Model Assessment (HOMA-BCF); +47.7+/-11.58%). Compared with placebo, fasting serum Tg concentrations decreased (-16.6%; P = 0.0178) and HDL-C concentrations increased (+12.6%; P= 0.0065) with pioglitazone as monotherapy. Total cholesterol and LDL-C changes were not different from placebo. The overall adverse event profile of pioglitazone was similar to that of placebo, with no evidence of drug-induced elevations of serum alanine transaminase (ALT) concentrations or hepatotoxicity. CONCLUSIONS: Pioglitazone improved insulin resistance and glycemic control, as well as Tg and HDL-C - which suggests that pioglitazone may reduce cardiovascular risk for patients with type 2 diabetes. PMID- 11491209 TI - NRA adverse reactions report. PMID- 11491208 TI - Bibliography. Current world literature. PMID- 11491210 TI - Members should ratify policies. PMID- 11491211 TI - FMD fight justifies the horror. PMID- 11491212 TI - Welfare: much more than science. PMID- 11491213 TI - Petalia websites--not for me! PMID- 11491214 TI - Vet pathology is disappearing. PMID- 11491215 TI - Reactions to tick antitoxin serum and the role of atropine in treatment of dogs and cats with tick paralysis caused by Ixodes holocyclus: a pilot survey. AB - OBJECTIVE: To determine the incidence and nature of adverse reactions of dogs and cats to tick antitoxin serum and to re-evaluate the role of atropine in the treatment of tick paralysis. DESIGN: A retrospective questionnaire of veterinarians. PROCEDURE: Questionnaires were posted to 320 veterinarians in tick endemic regions of Australia. Questions referred to dogs and cats treated for tick paralysis over a period of three years: the number treated, treatment protocols and adverse systemic reactions to tick antitoxin serum. Ninety completed questionnaires were returned and responses analysed. RESULTS: Veterinarians reported that approximately 3% of dogs exhibited adverse reactions immediately following treatment with tick antitoxin serum. Eighteen percent of these reactions were described as anaphylaxis, with the remaining 82% attributed to the Bezold-Jarisch reflex. Six percent of cats treated with tick antitoxin serum reacted adversely and the majority of reactions (63%) were ascribed to the Bezold-Jarisch reflex. Atropine was used routinely by 10% of responding veterinarians in the treatment of dogs and cats with tick paralysis. A similar number of veterinarians used atropine only in selected cases. Most veterinarians (76%) reported that they never used atropine in the treatment of tick paralysis in either dogs or cats. Within the survey population, premedication with atropine reduced the number of Bezold-Jarisch reactions following tick antitoxin administration approximately five-fold in dogs and four-fold in cats. CONCLUSIONS: Data from this pilot survey indicate that more cats than dogs have adverse systemic reactions to tick antitoxin serum and that the majority of these reactions in both dogs and cats could be related to the Bezold-Jarisch reflex. The number of reactions to tick antitoxin serum in dogs and cats could be significantly reduced by the routine use of atropine prior to administration of tick antitoxin serum. PMID- 11491216 TI - Thoracic actinomycosis (arcanobacteriosis) or nocardiosis causing thoracic pyogranuloma formation in three dogs. AB - OBJECTIVE: To describe three cases of canine thoracic actinomycosis (arcanobacteriosis) or nocardiosis in which the primary pathological lesion was a pyogranulomatous abscess in the mediastinum. Clinical signs, difficulties in diagnosis, treatment and prognosis are examined. Comparisons are made between human and veterinary literature to assist in formulating a rational treatment plan. DESIGN: Retrospective clinical study. PROCEDURE: Review of case records from 1984 to 1998. RESULTS: Three dogs presented with large intrathoracic pyogranulomas producing variable clinical signs, not necessarily associated with the respiratory tract. Ages ranged from 2 to 5 years old. Two dogs responded to surgical opening and passive drainage of the abscess, or surgical excision of the granuloma with associated structures, and medical therapy. One dog died intra operatively. CONCLUSION: A combination of surgical and antimicrobial therapy may carry a fair-to-good prognosis for thoracic granuloma caused by actinomycosis (arcanobacteriosis) or nocardiosis. The extent of surgery should be based on assessment of individual cases and must include surgical biopsy for histology and culture to enable a specific diagnosis to be made. Complete surgical excision is not necessarily required. Prolonged antimicrobial therapy is indicated. PMID- 11491217 TI - Defaecation syncope and pulmonary thromboembolism in a cat. AB - A 7-year-old male neutered domestic shorthair cat was referred for worsening gastrointestinal and haematologic abnormalities. Physical status deteriorated further despite intravenous crystalloids, blood transfusion and nutritional support. Cardiorespiratory signs developed and the cat died suddenly while straining to defaecate. Diffuse thrombosis, pulmonary thromboembolism, metastatic pancreatic carcinoma and histologic evidence of cardiomyopathy were present at necropsy. This is the first reported case of feline pulmonary thromboembolism associated with defaecation syncope. Predisposing factors for thrombotic disease in this case and aspects of human defaecation syncope are discussed. The risk of clot dislodgement by the Valsalva manoeuvre in patients with a thrombotic tendency is highlighted. PMID- 11491218 TI - Simian immunodeficiency virus infections in vervet monkeys (Clorocebus aethiops) at an Australian zoo. AB - A number of monkey species, including African green monkeys and African vervet monkeys (Chlorocebus aethiops), are frequently infected in the wild and in captivity with a Simian immunodeficiency virus strain, SIVagm, a primate lentivirus. Up to 50% of African green monkeys are estimated to be infected with SIVagm. SIV strains are very closely related to HIV-2 strains, which are a cause of AIDS in humans, predominantly in western Africa, although cases in Australia have also been reported. It is generally thought that SIV is non-pathogenic in several natural hosts, including African green monkeys. Nevertheless many SIV strains induce a profound immunodeficiency virtually identical to HIV-1 induced AIDS in humans when administered to Asian macaque species such as rhesus (Macaca mulatta) or pigtailed macaques (M nemestrina). SIV infection of Asian macaque species is frequently employed as an animal model for AIDS vaccine studies. In November 1996 a group of 10 African vervet monkeys were imported from the USA for display at Victoria's Open Range Zoo in Werribee. Two animals in this group of monkeys later developed a fatal gastroenteric illness. These diagnoses led us to initiate SIV testing of the colony. PMID- 11491219 TI - Acute pancreatitis in parrots. AB - Acute pancreatitis was diagnosed in three parrots. Antemortem diagnosis of pancreatitis in the avian patient is rarely documented, and should be suspected in birds showing clinical signs attributable to abdominal pain or gastro intestinal dysfunction. However, not all birds with confirmed pancreatitis display these signs. A serum amylase level greater than 1,500 U/L is suggestive of pancreatitis and pancreatic biopsy can be used to confirm a diagnosis. The aetiology of acute pancreatitis in parrots is discussed and a suggested treatment protocol is described. PMID- 11491220 TI - Prospective survey of tick paralysis in dogs. AB - OBJECTIVE: To obtain information on tick paralysis in dogs, including the nature of disease, host signalment, tick-host relationship, treatment, disease progression and recovery, and preventive measures. DESIGN: A prospective survey of 577 dogs affected by tick paralysis was conducted during 1998. Forty-two veterinary clinics along the eastern coast of Australia were instructed to complete survey forms for the first 15 dogs that presented with tick paralysis during September to November. RESULTS: Five percent of dogs died from tick paralysis. Younger dogs were more likely to survive. Long coat length was associated with a greater tick burden but not greater tick size, whereas coat thickness had no bearing on either. Dogs with mild disease recovered more quickly from tick paralysis. Respiratory and gait scores reflected disease severity and were good prognostic indicators. The size of the tick did not reflect the severity of the clinical condition it induced in the host. No method of tick removal or in situ treatment improved recovery time or reduced mortality. However, the time spent in hospital was significantly less for dogs from which the live tick was manually removed. Inspiratory stridor, evident in some dogs with tick paralysis, was not related to tick attachment on the neck. The use of acepromazine maleate or dexamethasone did not reduce recovery time or mortality. Increasing the dose of tick antitoxin serum (TAS) above 0.1 mL/kg had no effect on mortality or recovery time. Dogs with severe disease that received an additional dose of TAS were significantly less likely to survive. Subcutaneous use of TAS at the site of tick attachment was of no benefit in reducing mortality or time to initial clinical improvement. A registered preventative product had not been used on the majority of dogs. Clipping the coat to search for ticks did not reduce mortality. CONCLUSIONS: Therapy needs to address cardiopulmonary dysfunction that may be due directly to the effect of tick toxin and not just respiratory compromise caused by progressive respiratory muscle failure. PMID- 11491221 TI - 1,25(OH)2 vitamin D concentration in the plasma of Solanum glaucophyllum intoxicated rabbits. AB - OBJECTIVE: To test Solanum glaucophyllum calcinotic effects in adult New Zealand White rabbits in relation to cumulative dose and active principle concentration in plasma. DESIGN: An intoxication assay with controls. PROCEDURE: Rabbits were orally dosed with aqueous extracts of dry leaves of S glaucophyllum for 5, 7 or 9 days. During the experiment, body weight, calcaemia and phosphataemia were measured; retinal blood vessel calibre was observed by ophthalmoscopic examination of the ocular fundus. 1,25(OH)2 vitamin D plasma concentration was determined at the end of the experimental periods. Soft tissue calcium concentration and the presence of calcinotic lesions were studied after euthanasia. RESULTS: Toxic effects were evident in S glaucophyllum treated groups (loss of body weight, elevation of soft tissue calcium concentration, and presence of calcinotic lesions). Plasma 1,25(OH)2 vitamin D concentrations were negatively correlated with final body weight (r = -0.97; P < or = 0.001), and positively correlated with renal calcium concentration (r = 0.74; P = 0.02). There was also a significant regression of plasma 1,25 (OH)2 vitamin D concentration on the cumulative dose of S glaucophyllum (R2 = 0.87; P < or = 0.001). CONCLUSIONS: The procedure described here offers a sensitive and practical experimental model for the study of the pathogenesis of enteque seco. PMID- 11491222 TI - Serological examination for evidence of infection with Hendra and Nipah viruses in Queensland piggeries. AB - OBJECTIVE: To examine piggeries in Queensland for evidence of infection with Hendra virus and Nipah virus. DESIGN: A serological survey was designed to provide 99% confidence of detecting at least one infected pig herd in Queensland, assuming that for each virus, at least 5% of herds would have been exposed to virus and that at least 40% of the finisher pigs in these herds would have detectable antibodies to virus. PROCEDURE: A two stage sampling regimen was used. All samples were tested with serum neutralisation tests developed and performed at the Australian Animal Health Laboratory. RESULTS: There was no evidence of antibody to either virus in the 500 samples collected from 100 herds. CONCLUSION: The results of the survey support a case that commercial pigs in Queensland are free of both Hendra virus and Nipah virus infections. PMID- 11491223 TI - A neurological locomotor disorder in sheep grazing Stachys arvensis. AB - OBJECTIVE: To describe the clinical signs and pathology of a neurological locomotor disorder in sheep associated with ingestion of Stachys arvensis. DESIGN: Field observations and laboratory examinations. PROCEDURE: Clinical and pathological examinations were performed on sheep with suspected S arvensis intoxication. FIELD OBSERVATIONS: Merino sheep in a flock on the southwestern slopes of New South Wales developed a neurological disorder after grazing S arvensis. Affected animals had pelvic limb paresis and a proprioceptive deficit. When forced to exercise they stumbled and collapsed. Recovery following removal from the plant was slow. Many deaths were associated with the outbreak. RESULTS: Affected sheep developed a mild degenerative myelopathy and peripheral neuropathy. Deficiencies of vitamins A and E were also observed in the affected flock. CONCLUSION: Grazing S arvensis is sometimes associated with a neurological locomotor disorder in sheep. PMID- 11491224 TI - Eradication of ovine footrot by repeated daily footbathing in a solution of zinc sulphate with surfactant. AB - OBJECTIVE: To investigate the effect on ovine footrot of repeated daily footbathing in a solution of zinc sulphate with surfactant. DESIGN: Merino sheep were allocated to control and treatment groups of 119 sheep each at week 0. The sheep had a history of S1, U1, T and/or U6 types of Dichelobacter nodosus in interdigital and underrunning footrot lesions. Feet were not pared prior to treatment. PROCEDURE: Treatment sheep were footbathed in a 15 to 18% (w/v) solution of zinc sulphate with surfactant for 10 min on five consecutive days during week 1. At week 2, and fortnightly to week 52, all feet were inspected, lesion scores were recorded and samples were taken for laboratory tests. At week 53, all feet with no lesions at week 52, but with underrunning lesions prior to week 1, were pared and samples were taken. RESULTS: After footbathing, there were no lesions in any treatment sheep at any inspection to week 52. The percentage of feet of control sheep with lesions increased from 9% (391 of 4,284) between weeks 20 and 36, to 14% (593 of 4,284) between weeks 36 and 52. Ninety-five of 96 control sheep with no lesions at week 20 were still asymptomatic at week 52. D nodosus was not isolated from samples taken from 99 and 87 pared feet of treatment and control sheep, respectively. CONCLUSION: Repeated daily footbathing combined with prolonged exposure to a dry environment eradicated footrot in sheep with both interdigital and underrunning lesions in feet that were not pared prior to treatment. PMID- 11491225 TI - Cupped lesions of early onset dental erosion in young southeast Queensland adults. AB - BACKGROUND: Dental erosion manifests as cupped lesions on cusp apices and in fissures of teeth in patients from southeast Queensland referred with excessive tooth wear. When found in young adults, these lesions may indicate early onset of active dental erosion. If the numbers and extent of cupped lesions increase with age, erosion may be a slow cumulative process. METHODS: This cross-sectional study recorded the presence or absence and the relative sizes of cupped lesions from all cusps and occlusal fissures on premolar and permanent molar teeth from study models by image analysis. Type-specimens of cupped lesions were examined. RESULTS: The incidence by tooth reflected time in the mouth, post-tooth emergence. A linear increase in lesion number and size, with age, was found. However, cupped lesions occurred on mandibular first molar cusp apices as often, and attained greater extent, in adults under 27 years compared with older subjects. CONCLUSION: Marked differences were found between lesion number and size, between maxillary and mandibular molar sites that reflect differences in salivary protection against dental erosion. The significance of this study is that the mandibular first permanent molar indicates the age of onset and severity of dental erosion. PMID- 11491226 TI - A retrospective analysis of oral hairy leukoplakia in South Australia. AB - BACKGROUND: The features of oral hairy leukoplakia (OHL) have been widely reported in the literature. However, no studies have described this lesion in the Australian setting. This study retrospectively examines, with respect to specific clinical factors, the prevalence of OHL in a South Australian HIV-infected population. METHODS: Clinical data were collected from the records of 197 HIV infected patients who had attended the Adelaide Dental Hospital between January 1986 and February 1995. Data were analysed using the chi-square test. RESULTS: The prevalence of OHL in South Australian HIV-infected patients was 45.2 per cent. The study found the presence of OHL was not related to CD4+ T-lymphocyte count or AIDS-defining illness nor did the length of time a patient had been infected with HIV relate to the presence of OHL. An association was observed between a reduced prevalence of OHL in patients who were taking antiviral medication. CONCLUSION: The prevalence of OHL in South Australia is comparable with results of other studies. This study supports the notion that OHL is not an indicator of immunosuppression in South Australian HIV-infected patients. Further longitudinal studies are required to ascertain the relationship of OHL to HIV disease progression. PMID- 11491227 TI - Prevalence of periodontal conditions among public-funded dental patients in Australia. AB - BACKGROUND: The aims of this study were to describe periodontal health status by gender, type of care, geographic location and age and to investigate associations of these factors with tooth loss. METHODS: In 1995-96, state and territory dental services surveyed a random sample of 6,109 public-funded patients. Dentists recorded oral health status at the initial visit of a course of care, using written instructions but without formal calibration. Periodontal status was assessed using the community periodontal index of treatment needs. RESULTS: Prevalence of periodontal conditions among dentate patients was higher among older, male, emergency and urban patients (chi2; p<0.05). Age-specific edentulism was lower for male, emergency and urban patients (chi2; p<0.05). Numbers of missing teeth were lower for younger, non-emergency and urban patients (Anova; p<0.05). Prevalence of periodontal pockets 6mm or more was associated with gender, type of care, geographic location, age and number of missing teeth (logistic regression; p<0.05). CONCLUSIONS: Prevalence of severe periodontal conditions was lower in 1995-96 compared with public-funded patients in 1984 and 1992-93, but prevalence of periodontal health remained lower than the general population in 1987-88. A survivor effect may result in lower recording of severe periodontal conditions among subgroups with higher tooth loss. PMID- 11491228 TI - Patterns of access to public oral health care in Queensland by gender, indigenous status and rurality. AB - Data were collected on courses of care from all attendances at public sector dental clinics between July 1994 and June 1996 and examined by gender, rurality and indigenous status. Males and indigenous people were more likely to attend dental clinics when a problem with pain existed, resulting in a high need for emergency services, more diagnoses of dental caries, and treatment more often involving oral surgery, including extractions. Similarly, remote area dwellers had more diagnoses of dental caries and oral surgery, although their attendance at clinics was complicated by access. PMID- 11491229 TI - Ascending necrotising fasciitis as a result of odontogenic infection: a report of two cases. AB - Necrotising fasciitis is a severe soft tissue infection which spreads rapidly through fascial planes, is characterised by soft tissue necrosis and is potentially life-threatening. It is a rare entity in the head and neck region. The management of this condition is difficult and early diagnosis and aggressive surgical and medical management are essential. This paper reports two cases of necrotising fasciitis as a result of ascending odontogenic infection involving the temporalis muscle. PMID- 11491230 TI - Continuing education: the 1998 Survey of the Royal Australasian College of Dental Surgeons. AB - BACKGROUND: Continuing education (CE) is an essential professional activity. In the last decade, CE has been actively pursued by the medical profession in Australia and abroad. However, the uptake of CE in dentistry has been much slower and there is minimal Australian data on dental CE. METHODS: To determine the level of CE activity, in 1998, postal questionnaires were sent to all fellows of the Royal Australasian College of Dental Surgeons. The responses were analysed. RESULTS: There was a high response rate (90 per cent) but a moderate usable rate (54 per cent). The results show a biphasic distribution between high and low CE activity. The average amount of activity of those involved in CE was 116 hours per year, above the usually accepted minimum of 100 hours/year. Some groups, particularly members of the specialist divisions of oral and maxillofacial surgeons (215 hours) and periodontists (205 hours), have high levels of CE. However, approximately 25 per cent of college fellows reported little or no CE activity. The survey revealed that inactive fellows are more likely to be older and in general practice. Inactive fellows were also tardy in replying to the questionnaire. CONCLUSION: The high activity CE group needs to be recognised and encouraged to continue. Specific plans to help the low CE activity group should be developed. Although these findings relate directly to the Royal Australasian College of Dental Surgeons, they are presented as they have implications for the dental profession at large. PMID- 11491231 TI - Did dental flossing cause endocarditis? PMID- 11491232 TI - The hazards of radiography. PMID- 11491233 TI - Oral health report. PMID- 11491234 TI - Full occlusal protection--theory and practice of occlusal therapy. AB - In this review the full occlusal protection theory is proposed and its clinical practice demonstrated for varying Angle's malocclusions. The concept of developing restorative anatomical shapes or bio-designing occlusal schemes of crowns to compensate for poor intra-arch tooth positioning is proposed and demonstrated. Clinical parameters for full occlusal protection in occlusal therapy are presented. PMID- 11491235 TI - Water fluoridation, osteoporosis, fractures--recent developments. AB - BACKGROUND: Optimal (1ppm) water fluoridation is seen as the most socially equitable way to prevent dental caries, however concerns about the safety of fluoridation are periodically raised. METHODS: Research on effects on bone published since the 1991 National Health and Medical Research Council report on water fluoridation was reviewed. RESULTS: Thirty-three studies were identified. Adverse effects in animal feeding studies were only seen at doses much greater than those currently used in artificial water fluoridation. The majority of animal studies showed no effect or a beneficial effect of low fluoride doses. The results of ecological studies were conflicting. One of the two cohort studies showed an increase in fracture incidence at fluoride levels four times greater than optimal water fluoridation and the other showed no effect after 20 years' optimal fluoridation. The cross-sectional studies showed a favourable effect on bone mineral density. The clinical trials predominantly showed increased bone density in several sites associated with fluoride treatment of 9-22.6mg fluoride per day for one-four years. CONCLUSION: These studies provide a substantial body of evidence that fluoride at up to 1ppm does not have an adverse effect on bone strength, bone mineral density or fracture incidence. PMID- 11491236 TI - Dental complications of head and neck radiotherapy: Part 1. AB - Radiotherapy for head and neck tumours is a viable treatment modality. However, a wide range of potentially debilitating dental complications may accompany this treatment. The nature and impact of these complications are outlined in this first part of a two-part article. In Part 2, prevention and management strategies available to the dental practitioner to stave off the dental side effects of radiotherapy will be explored. PMID- 11491237 TI - A comparison of Kodak Ultraspeed and Ektaspeed Plus dental X-ray films for use in endodontics. AB - BACKGROUND: The advantage of using a faster film for length determination in endodontic therapy is obvious. However, for such a film to be generally accepted, it must demonstrate comparable diagnostic quality to traditionally used films. METHODS: The comparative accuracy of canal length determination of Ultraspeed and Ektaspeed Plus dental X-ray films was assessed in maxillary first and second molars; for different canals, for different teeth, for different exposures, and for different examiners (five general dentists and three endodontic specialists). RESULTS: In general, there were no significant differences between films, among examiners, or any interaction between films and exposures. That is, an assessor's ability to estimate lengths was not significantly influenced by the film type or by exposure used. There was a wide divergence in the individual assessor's ability to estimate lengths. Specialists estimated lengths more accurately than general practitioners and estimated lengths more accurately with Ektaspeed Plus film. Length determination in distobuccal and mesiobuccal canals was more accurate than in palatal canals. Most palatal canals were underestimated in length by more than 1mm. The use of file sizes larger in number than size 15 is recommended in these canals. CONCLUSION: For length determination, Ektaspeed Plus dental X-ray film is as effective as Ultraspeed film. Given the acceptable quality and accuracy of Ektaspeed Plus film, there seems to be no clinical reason to subject patients to greater radiation by using a slower film during endodontic therapy. PMID- 11491238 TI - The effects of introducing or lowering legal per se blood alcohol limits for driving: an international review. AB - In this review evidence on the impact of introducing or lowering legal blood alcohol limits on traffic safety measures is examined. There is substantial variability in the types and rigour of methods used to evaluate these legislative measures, and thus not surprisingly there is variability in the results observed. In most but not all cases where an evaluation of an introduced or lowered legal limit has been conducted, some beneficial effect on traffic safety measures has been reported. These effects are in some cases relatively small, and in other cases may be temporary. In some jurisdictions, lasting reductions in collision rates have been reported. Available evidence suggests that where beneficial effects are observed they are due to general deterrence, and not restricted only to drivers at blood alcohol concentrations (BAC) specifically affected by the legal change. PMID- 11491239 TI - Do speed bumps really decrease traffic speed? An Italian experience. AB - Italy introduced the extensive use of speed bumps only in 1990, in an attempt to limit the high number of fatalities involving pedestrians in urban streets caused by the high speed of vehicles. In many countries, such devices have been the subject of careful investigations (in order to assess their effectiveness and disadvantages for the traffic circulation) and this has resulted in a number of modifications in the design to improve their performance. On the contrary, no systematic and scientific studies have been carried out on Italian installations: moreover, the type of undulation adopted is known to produce a series of problems for some categories of users and is not so effective in reducing speed as larger devices such as 'speed humps' or 'speed cushions'. This paper proposes a study of the effectiveness of 23 speed bumps installed in the city of Cagliari; to this aim, a speed analysis was performed at speed bump locations, at the crosswalks protected by the devices and at sections of the streets where bumps are installed but far from them. The results show that in one third of the cases the 85th percentile of speed measured at the speed bumps is higher than the posted speed limit (50 km/h) and an equal percentage of vehicles travel at a speed in the range of 45-50 km/h. No statistically significant differences were found from the comparison of speed values observed in free, bump or crosswalk sections of the same streets, while speed profiles calculated at four sites, where a high percentage of braking vehicles was observed, showed a common trend from which it clearly emerges that the effect of the device on driver's behaviour is restricted to a short spatial range (about 20-30 m before and after the bump). The current situation thus suggests the use of more effective devices such as humps or cushions, or the integration of speed bumps with other traffic calming techniques. PMID- 11491240 TI - Evaluation of inter-organizational traffic injury prevention in a WHO safe community. AB - The objective of the study was to examine the effect of a community-based injury prevention program on traffic injuries. A population-based quasi-experimental design was used with pre- and post-implementation measurements in an intervention and a control area. The program was based on inter-organizational participation in detecting and taking action against traffic injuries. The total relative risk for traffic injury in the study area showed only a tendency to decrease following program exposure (odds ratio 0.91; 95% confidence interval 0.81-1.02). No change in relative risk was observed in the control area. The analyses of program impact on injury severity showed that the relative risk for moderate injuries in the study area was reduced by almost half (odds ratio 0.59; 95% confidence interval 0.49-0.69), the risk for severe or fatal injuries remained constant (odds ratio 1.27; 95% confidence interval 0.80-2.02), and the risk for minor injuries increased (odds ratio 1.34; 95% confidence interval 1.13-1.59). The relative risk for moderate injuries was reduced by at least half for mopedists, cyclists, pedestrians, and those leaving or entering a motor vehicle. Community-based injury prevention can be a complement to national traffic safety programs. PMID- 11491241 TI - Predicting repeat DUI offenses with the alcohol interlock recorder. AB - The aim of this report has been to use information contained in the alcohol ignition interlock recorder to determine whether systematic analysis of it can be used to predict which DUI offenders will recidivate during the first 2 years after the interlock is removed. The interlock record was accumulated during a 4 year intervention study in Alberta, Canada. Data from more than 5.5 million breath tests collected during interlock use were analyzed retrospectively after allowing repeat DUI offenses to accumulate for up to 2 years post-interlock. The rate of interlock warns at low BAC (0.02-0.04%) and fails at higher BAC ( > 0.04%) were found to be predictive of later repeat DUI. The interlock record was used along with selected driver record variables and questionnaire data to identify predictor sets. CHAID segmentation analysis was used to identify combinations of predictor variables; these were joined with sensitivity analysis to compare different predictor combinations. Several variables, but primarily more prior DUIs and more interlock warns and fails logged during the first 5 months of interlock usage predict greater than 60% of repeat DUI with a false positive rate of less than 10%. PMID- 11491242 TI - The development of a test specification to determine the rollover protection of passengers in light commercial vehicles fitted with canopies. AB - In this paper, a basic test specification is proposed to assess the structural integrity of a canopy that covers the load-area of a light commercial vehicle (LCV) used to carry passengers in the rear. The proposed test specification is a first step towards improving the safety of these passengers during rollover accidents. Different test methods were evaluated for applicability, ease of use and financial implications. The mass and geometric characteristics of three different one-ton LCVs were experimentally obtained and used to determine the minimum lateral velocity that will result in a rollover should a tripping mechanism be encountered. These velocities, in conjunction with other published data, were used to determine the energy required for a pendulum qualification test. It was decided that 7.7 kJ of energy delivered by a pendulum travelling between 4 and 7 m/s just prior to impact would be the most representative load case. A finite element analysis (FEA) of the proposed impact test on a specimen canopy was completed. Thereafter, the actual pendulum impact test was conducted to demonstrate the applicability of the proposed test procedure as well as the correlation with the FEA. It was shown that both FEA and actual testing are viable qualification procedures and that they can in fact be used to reliably assess the strength of the canopy. Finally, the outline of a test specification was developed and proposed, which can be used by national regulatory institutions to assess the structural integrity of commercially manufactured canopies. PMID- 11491243 TI - Drivers' biased perceptions of speed and safety campaign messages. AB - One hundred and thirteen drivers were surveyed for their perceptions of driving speed to compare self-reported average speed, perceived average-other speed and the actual average speed, in two conditions (50 and 100 kph zones). These contrasts were used to evaluate whether public safety messages concerning speeding effectively reach their target audience. Evidence is presented supporting the hypothesis that drivers who have a biased perception of their own speed relative to others are more likely to ignore advertising campaigns encouraging people not to speed. A method of self-other-actual comparisons detects biased perceptions when the standard method of self-other comparison does not. In particular, drivers exaggerate the perceived speed of others and this fact is masked using traditional methods. The method of manipulation is proposed as a way to evaluate the effect of future advertising campaigns, and a strategy for such campaigns is proposed based on the results of the self-other comparisons. PMID- 11491244 TI - Helmet laws and motorcycle rider death rates. AB - We investigated motorcycle rider death rates between states with full motorcycle helmet laws and those without. This was done using both unadjusted bivariate analyses and multivariate random-effects generalized least squares regression models of rider death rates. Multivariate models were adjusted for the competing influences of several explanatory variables, including the existence of a motorcycle helmet law. From 1994 to 1996, states with helmet laws experienced a median death rate of 6.20 riders per 10000 registered motorcycles and states without helmet laws experienced a median death rate of 5.07 riders per 10000 registered motorcycles (P = 0.008). After controlling for other factors that affect motorcycle rider fatalities (most notably population density and temperature), death rates in states with full helmet laws were shown to be lower on average than deaths rates in states without full helmet laws (P = 0.740). Our study weakens the claim that rider death rates are significantly lower in states without full motorcycle helmet laws. PMID- 11491245 TI - Adolescent antecedents of high-risk driving behavior into young adulthood: substance use and parental influences. AB - Driver history data, in combination with previously collected tenth-grade questionnaire data, for 4403 subjects were analyzed by Poisson regression models to identify the significant substance use and parental characteristics predicting subsequent high-risk driving of new drivers (starting at age 16) through age 23 24 years. Substance use (cigarettes, marijuana, and alcohol) reported at age 15 was shown to be an important predictor of subsequent excess risk of serious offenses and serious crashes for both men and women. In addition, negative parental influences (lenient attitudes toward young people's drinking; low monitoring, nurturance, family connectedness), were also demonstrated to increase the risk of serious offenses and serious crashes for both men and women. PMID- 11491246 TI - The influence of demographic factors on seatbelt use by adults injured in motor vehicle crashes. AB - This study determined demographic factors associated with reported seatbelt use among injured adults admitted to a trauma center. A retrospective chart review was conducted including all patients admitted to a trauma center for injuries from motor vehicle crashes (MVC). E-codes (i.e. ICD-9 external cause of injury codes) were used to identify all patients injured in a MVC between January 1995 and December 1997. Age, sex, race, residence zip code (i.e. a proxy for income based on geographic location of residence), position in the vehicle, and seatbelt use were obtained from the trauma registry. Forward logistic regression was used to identify significant predictors of seatbelt use. Complete data was available for 1366 (82%) patients. Seatbelt use was reported for 45% of patients under age of 25 years, 52% of those 25-60 years, and 68% of those over 60 years. Overall, seatbelt use was reported for 45% of men and 63% of women, as well as for 56% of Caucasians (i.e. Whites) and 34% of African Americans. In addition, seatbelt use was reported for 33% of those earning less than $20,000 per year and 55% of those earning over $20,000. Finally, seatbelt use was reported for 57% of drivers and 43% of passengers. Logistic regression revealed that age, female gender, Caucasian race, natural log of income, and driver were all significant predictors of reported seatbelt use. These results show that seatbelt use was more likely to be reported for older persons, women, Caucasians, individuals with greater incomes, and drivers. Seatbelt use should be encouraged for everyone; however, young people, men, African Americans, individuals with lower incomes, and passengers should be targeted specifically. PMID- 11491247 TI - Explaining national road fatalities. AB - The purpose of this paper is to report on a model that can be used to explain the differences in road fatalities of individual countries. National infrastructure, transportation and socio-economic variables from international databases were considered as possible variables. The model was developed by means of stepwise regression analyses. It was found that the passenger car ownership is a better predictor of fatalities per 100000 passenger cars than vehicle ownership as a predictor of fatalities per 100000 vehicles. Many individual infrastructure and socio-economic variables have a significant effect on the fatality rate. The final model includes passenger car ownership, the Human Development Index (HDI), and the percentage of other vehicles as explanatory variables. PMID- 11491248 TI - Factors contributing to the amount of vehicular damage resulting from collisions between four-wheel drive vehicles and passenger cars. AB - This study examines the vehicular damage resulting from motor vehicle crashes involving four-wheel drive vehicles and passenger cars in the state of Oklahoma. In particular, the focus of the analysis is on differences in vehicular damage to passenger cars and four-wheel drive vehicles. Results indicate that passenger cars sustain significantly greater vehicular damage than four-wheel drive vehicles. In addition, several other factors significantly influence the level of damage resulting from collisions between four-wheel drive vehicles and passenger cars. Driver behavior or unsafe acts, represented by the rate of travel prior to the collision, failure to yield, failure to obey a stoplight or a stop sign, the consumption of alcohol and the use of drugs, also contributed to the amount of vehicular damage. In addition, results also indicate that the level of damage was influenced significantly by environmental factors, represented by a reduced intensity of light, wet or slippery roadways and the type of collision. The findings reinforce the importance of a number of policy initiatives that may reduce the vehicular damage resulting from collisions involving four-wheel drive vehicles and passenger cars. For example, the study indicates a need to initiate legislation that lowers the speed limit during dark and twilight hours, commits additional resources to road maintenance to reduce unsafe road conditions, and stimulates improvements in automotive design that provide better lateral protection to vehicles. PMID- 11491249 TI - Sensation seeking, risky driving and behavioral adaptation. AB - A study on the relationship between sensation seeking (SS) and risky driving, aggressive driving and behavioral adaptation is reported on. College students completed a questionnaire concerning their driving behavior and level of sensation seeking. Results indicated that high SS's were significantly more likely than low SS's to speed, not wear belts, drink frequently, drive after drinking, perceive a low risk of detection for impaired driving, and perceive that they could drink more beer before being impaired. High SS's were also more likely to report aggressive driving habits. High SS's were significantly more likely than low SS's to say that they would drive faster on highways and on wet roads and drive after drinking, if operating a vehicle equipped with anti-lock brakes. The results are consistent with previous research. PMID- 11491250 TI - Putting head restraints to rest. PMID- 11491251 TI - Changes in head injury with the New Zealand bicycle helmet law. AB - It was claimed that the bicycle helmet law in New Zealand reduced head injuries to adult cyclists by 28% (Povey, L.J., Frith, W.J., Graham, P.G., 1999. Cycle helmet effectiveness in New Zealand. Accident Analysis and Prevention 31, 763 770). However, the pre-law increase in adults wearing helmets (from 30% in 1990 to 43% in 1993) was accompanied by a fall of 45 head injuries per 100 limb injuries (i.e. -3.47 for every 1% increase in helmet wearing) compared with a fall of 11 when wearing increased from 43 to 93% with the law (-0.23 for every 1% increase in wearing). Unless voluntary wearing is 15 times more effective in reducing head injuries, it seems likely that the apparent effects (as described by Povey et al., 1999) were an artefact caused by failure to fit time trends in their model. Such inconsistency of effects over periods of substantial change compared with periods of little change in helmet wearing may be a useful indicator of the presence of trends. Because the large increases in wearing with helmet laws have not resulted in any obvious change over and above existing trends, helmet laws and major helmet promotion campaigns are likely to prove less beneficial and less cost effective than proven road-safety measures, such as enforcement of speed limits and drink-driving laws, education of motorists and cyclists and treatment of accident black spots and known hazards for cyclists. PMID- 11491252 TI - Viewpoint. Environmental issues: insight into corresponding problems of traffic safety. PMID- 11491253 TI - Interview with Janusz Szajewski. AB - Janusz Szajewski, the founder of the Warsaw Poisons Control Centre and Haemodialysis Unit at Praski Hospital in Warsaw, Poland, was born on February 25, 1924 in Grodzisk, Poland. He undertook his undergraduate studies from 1939 to 1945 at the underground Chrobry College in Piotrkow, Poland, at a time when higher education was outlawed to Poles and punishable by imprisonment at Auschwitz. At war's end, Dr. Szajewski went to medical school at the Medical University of Warsaw, Warsaw, Poland, from 1945-1950. Further training in Internal Medicine was obtained at the Medical University of Warsaw. Because of his former activity in the underground Polish Army during the war his training was interrupted in 1951 by a 6-month deportation by the Polish communist regime to Silesia andforced employment as a coal-mine physician. During the 1950s, he also worked as chief of the Department of Internal Medicine in the Polish Red Cross Hospital in Hamhyng, North Korea. Subsequently, from 1959-1960, Dr. Szajewski served as a Rockefeller Fellow at the Cleveland Clinic Foundation, under the guidance of Irvine H. Page. Dr. Szajewski began his work with the Warsaw Poisons Control Centre and Haemodialysis Unit in 1970 and directed the center 1970-1996. PMID- 11491254 TI - Risk assessment by physicians and by the poison center: are nondrug exposures handled too actively? AB - BACKGROUND: The study was intended to evaluate the possible impact of poison center advice on physicians' treatment and referral of nondrug exposures. METHODS: Doctors seeking advice on nondrug poisoning were asked for their management plan before any recommendations were given. The proportions of cases in different treatment and referral categories were compared to the poison center recommendations. Rate ratios were used as effect measure. RESULTS: A total of 175 cases were included in the study. For 90% of these, the exposure was estimated by the poison center to be of no or minor risk. The inquiring physicians had intended to treat 43 more cases than the poison center recommended, rate ratios: 1.6 (95% CI: 1.3-2.0). The most marked difference wasfor treatment by gastrointestinal decontamination, rate ratios: 5.0 (95% CI: 2.5-9.8). For referral, 42 more cases would have been observed in a stationary hospital unit, rate ratios: 2.3 (95% CI: 1.6-3.3) and 28 excess cases would have been observed in an outpatientfacility, rate ratios: 2.0 (95% CI: 1.3-1.6). A more active attitudefor treatment and referral by the inquirers was independently associated with patients < or =14 years of age, lack of symptoms at presentation, and insignificant exposures. CONCLUSIONS: Doctors seeking advice on the management of nondrug poisoning intended to treat and refer to health care facilities more actively than recommended by the poison center. The tendency was most pronouncedfor children exposed to substances of minor toxicological significance and treatment with gastrointestinal decontamination. PMID- 11491255 TI - Parent and caregiver compliance with poison center advice. AB - BACKGROUND: The purpose of this study was to assess the compliance of parents and caregivers whose children (under 6 years of age) accidentally ingested antihistamines, decongestants, or both. METHODS: We used a prospective descriptive study design and developed a convenience sample from eligible calls retrieved from the records from January 19, 1999 to February 28, 1999. Poison control specialists categorized the children into two groups: toxic and needing treatment in a hospital or nontoxic and not needing treatment at a hospital. Compliance with Poison Control Center advice was measured by follow-up telephone calls made by a research assistant within 7 days of the initial call. RESULTS: One hundred forty-seven calls were retrieved and followed. One hundred and seven (72.8%) callers had been advised that no treatment was needed. One hundred (93.4%) of these 107 followed Poison Control Centre advice. Forty (27.2%) of the 147 callers were advised to take their child to an emergency department. Thirty nine (97.5%) of these 40 complied with the advice. There was no statistically significant difference in compliance rates between the two groups (chi2 = 0.306, df = 1, p = 0.58). DISCUSSION AND CONCLUSIONS: In this group of callers, compliance was high. Reasons for noncompliance included caregiver anxiety, specifically the need for a second opinion. Future studies are needed to determine the level of compliance more definitively. Studies should include subjects from other age groups, other exposures, and other types of callers. PMID- 11491256 TI - Protected renal stenting with the PercuSurge GuardWire device: a pilot study. AB - PURPOSE: To evaluate the feasibility and safety of renal artery angioplasty and stenting utilizing a distal protection device to reduce the risk of intraprocedural atheroembolism. METHODS: Twenty-eight hypertensive patients (18 men; mean age 71.3 +/- 8.6 years, range 49-87) with atherosclerotic renal artery stenosis (4 bilateral) underwent angioplasty and stenting with distal protection in 32 renal arteries (29 ostial lesions). The lesion was crossed with a GuardWire temporary occlusion balloon, which was inflated to provide parenchymal protection. Generated debris was aspirated and analyzed. Blood pressure and serum creatinine levels were followed. RESULTS: Immediate technical success was 100%. All lesions were stented, either directly (14 ostial lesions), after predilation (15 ostial lesions), or owing to suboptimal angioplasty (3 nonostial lesions). Visible debris was aspirated from all patients. Mean particle number and diameter were 98.1 +/- 60.0 per procedure (range 13-208) and 201.2 +/- 76.0 microm (range 38-6206), respectively. Mean renal artery occlusion time was 6.55 +/- 2.46 min (range 2.29-13.21). Mean follow-up was 6.7 +/- 2.9 months (range 2-17). Systolic and diastolic blood pressure declined from 167.0 +/- 15.2 and 103.0 +/- 12.0 mm Hg, respectively, to 154.7 +/- 12.3 and 93.2 +/- 6.8 mm Hg after the procedure. The mean creatinine level dropped from 1.34 +/- 0.35 mg/dL preprocedurally to 1.22 +/- 0.36 mg/dL at 24 hours and remained constant. At 6-month follow-up, renal function did not deteriorate in any patient, whereas 5 patients with baseline renal insufficiency improved after the procedure. CONCLUSIONS: These preliminary results suggest the feasibility and safety of distal balloon occlusion during renal interventions to protect against atheroembolism. This technique's beneficial effects should be evaluated by randomized studies. PMID- 11491257 TI - Intravascular ultrasound-guided renal artery stenting. AB - PURPOSE: To evaluate the clinical outcomes of patients undergoing renal artery stenting with intravascular ultrasound (IVUS) guidance and compare measurements between IVUS and angiography. METHODS: One hundred thirty-one patients (71 women; mean age 71 +/- 8 years) underwent IVUS-guided Palmaz stent implantation in 153 stenotic renal arteries at a single center. The indications for stenting were uncontrolled hypertension (102, 77.9%), renal insufficiency (10, 7.6%), and both conditions (19, 14.5%). The majority of lesions were ostial (114, 74.5%); the remainder occupied the proximal renal artery (39, 25.5%). The mean lesion length and diameter stenosis were 6.5 +/- 3.0 mm and 74% +/- 10%, respectively, as measured by angiography. Data were recorded in a prespecified database; angiographic and IVUS images were analyzed at dedicated core laboratories and compared. RESULTS: Angiographic success was achieved in all patients, but IVUS indicated the need for additional intervention in 36 (23.5%) cases. There was strong correlation between the angiographic and IVUS measurements of lesion length (r = 0.60, p < 0.0001) and pre-/postprocedural minimal luminal diameter (r = 0.72 and 0.63, respectively; p < 0.0001). The mean contrast volume was 74 +/- 18 mL per case. In-hospital renal failure occurred in 8 (6.1%) patients; 2 (1.5%) required transient hemodialysis. At a mean 15-month follow-up, patients were treated with fewer antihypertensive medications (p = 0.05), and systolic and diastolic arterial blood pressures had decreased (p = 0.001); no significant change was noted in serum creatinine. CONCLUSIONS: IVUS-guided stenting facilitates safe renal artery revascularization. IVUS imaging may complement angiography in certain cases, which should be studied further in prospective studies with iodinated or noniodinated contrast agents. PMID- 11491258 TI - Corroded nitinol wires in explanted aortic endografts: an important mechanism of failure? AB - PURPOSE: To analyze surface alterations and fractures observed in the nitinol stent wires of explanted endovascular grafts used for treatment of abdominal aortic aneurysm. METHODS: Twenty-one explanted Stentor devices and 1 Cragg stent were received from investigators in Germany and France. After macroscopy and photography, the explants were cleaned and the polyester coating removed. The frame was examined completely by stereomicroscopy, and irregularities were assayed by scanning electron microscopy and energy dispersive x-ray analysis (EDAX). The observed alterations were classified according to stereomicroscopic and electron microscopic morphology. RESULTS: The mean implantation interval for the endografts was 29.1 +/- 13.2 months (range 5-46). All examined explants, even those retrieved after only a few months in situ, showed pitlike surface damage 10 25 microm in diameter. Larger, irregularly shaped surface alterations were observed in approximately 70% of the explants. Older explants (age >32 months) presented vast regions of decay, with bending of the wire and stress cracks in some areas. EDAX examination revealed decreased nickel concentration in the corroded regions. CONCLUSIONS: Corrosion of the nitinol wire in endovascular grafts is confirmed. Presumably, the observed pitting and irregularly shaped corrosion defects are the precursors of material failure. They weaken the thin wire, which leads to stress cracks and eventually fracture of the stent wire under circulatory pulsation. Cell-induced electrochemical corrosion and active cellular destruction of surfaces are well-known mechanisms that must be investigated for their possible roles in the corrosion of stent metals. PMID- 11491259 TI - Failed AAA endograft exclusion due to type II endoleak: explant analysis. AB - PURPOSE: To report the patient history and analysis of an explanted modular bifurcated endograft that was implanted to exclude an abdominal aortic aneurysm (AAA). CASE REPORT: An 80-year-old man with a 6-cm AAA underwent uneventful endovascular implantation of a bifurcated AneuRx stent-graft. His postprocedural clinical course was uneventful, although persistent contrast enhancement of the aneurysm remained via the inferior mesenteric artery (IMA). By 6 months, an endoleak connecting to the lumbar and mesenteric arteries became apparent. Over the ensuing 12 months, the endoleak and aneurysm enlarged; branch artery embolization was attempted in 4 percutaneous procedures. Despite successful IMA occlusion, the aneurysm continued to increase in diameter and volume, necessitating conversion to a conventional bypass at 20 months. Analysis of the explanted specimen revealed an intact endograft with fibrous incorporation of the stent framework at the proximal and distal fixation sites only; no incorporation of the endograft was noted within the aneurysm. The feeding channel for the endoleak was not identified. CONCLUSIONS: Serial imaging is a vital component of endograft surveillance, and persistent type II endoleaks that cannot be completely embolized endanger the longevity of the aneurysm exclusion. Explant analysis can play an important role in understanding the mechanisms of endograft failure. PMID- 11491260 TI - Endovascular grafting of traumatic aortic aneurysms in contaminated fields. AB - PURPOSE: To evaluate the potential of endovascular stent-grafts to treat traumatic aortic lesions in contaminated areas. METHODS: Four patients (3 women; ages 26-78 years) underwent stent-grafting to repair an aortic rupture sustained in a motorcycle accident, aortic lacerations secondary to surgical treatment of spondylitis in 2 patients, and an aortobronchial fistula following surgical thoracic aortic repair 10 years earlier. Stent-grafts (2 Corvita, 1 Talent, and 1 Vanguard) were placed endoluminally into the infected areas via a transfemoral approach. Follow-up included erythrocyte sedimentation rate, white blood count, C reactive protein, blood cultures, and computed tomography (CT). RESULTS: The stent-grafts were successfully placed in all cases and excluded the aortic lesion. Under supportive antibiotic therapy, inflammation parameters returned to normal. CT imaging showed no evidence of paraprosthetic infection, nor were there any other complications over a follow-up that ranged from 3 to 34 months. CONCLUSIONS: Endovascular therapy may be an alternative in the acute management of aortic ruptures in the setting of infection. Long-term results are required for definitive evaluation of the method. PMID- 11491261 TI - Intraoperative proximal endoleaks during AAA stent-graft repair: evaluation of risk factors and treatment with Palmaz stents. AB - PURPOSE: To study factors that might contribute to intraoperative proximal type I endoleak and to evaluate the placement of giant Palmaz stents as a therapeutic option. METHODS: Thirty-three patients (30 men; median age 72 years, range 50-85) with abdominal aortic aneurysms underwent implantation of fully supported Gianturco Z-stent-based endografts (12 custom-made aortomonoiliac and 21 bifurcated Zenith devices). Ten (30%) patients were treated for intraoperative proximal endoleaks. Stent-graft oversizing and neck angulation, length, and shape were compared between patients with and without leaks. RESULTS: In 9 cases, the endoleaks were successfully treated with intraoperative placement of Palmaz stents without complications. In 1 patient, a leak that was resolved intraoperatively with balloon dilation reappeared 1 month later; a Palmaz stent was deployed successfully. Stent-graft oversizing did not differ significantly between patients who developed proximal endoleaks and those who did not (median 4.0 mm in both groups, p = 0.47). Median neck length was 21.0 mm in patients with endoleak and 28.0 mm in those without (p > 0.99). Median neck angulation was 30 degrees in both groups (p = 0.33), and the presence of a conical aneurysm neck was not significantly different (2/10 versus 6/23, p > 0.99). All aneurysms remained excluded at a median follow-up of 13 months (range 6-24). CONCLUSIONS: Stent-graft oversizing and neck morphology (length, angulation, and conical shape) do not seem to correlate with the incidence of proximal type I endoleaks. Palmaz stent placement appears to be a feasible and safe treatment option for this complication. PMID- 11491262 TI - Healing response of normal canine aorta and iliac artery to a nitinol stent encapsulated in carbon-lined ePTFE. AB - PURPOSE: To evaluate the healing response of normal canine arteries to a self expanding nitinol stent encapsulated in carbon-lined expanded polytetrafluoroethylene (ePTFE). METHODS: Twenty-eight dogs were divided into aortic (n = 18) or iliac (n = 10) groups. In the latter, 2 animals were assigned to implantation intervals of 7, 30, and 90 days, respectively; 4 were designated for 180-day implantation. Half of the animals in each subgroup received a second overlapping stent-graft in one iliac artery. In the aortic cohort, 6 animals were assigned to the 180-day implantation group (2 with dual devices) and 3 to each of the others (1 dual implantation in each group). The devices were evaluated with angiography and intravascular ultrasound at implantation and explantation. After harvesting and gross examination, the specimens were examined microscopically and with scanning electron microscopy. RESULTS: The 49 implanted devices (24 aortic and 25 common iliac) were all widely patent at explantation, save for 2 iliac stents that had moderate (<40%) stenosis. No neointima was present at the 7-day interval. All stents were covered by thin neointima (<150 microm) at 30 days. At 180 days, an endothelial lining was present in the proximal and distal segments of all stents; in 4 of the 6 aortic stents, this endothelial lining was complete, whereas none of the iliac devices had endothelium in the midsegment at 180 days. At 1 year, 2 of the aortic specimens had an incomplete endothelial lining, whereas the lining was complete in the third. There was no evidence of stent graft migration or inflammation associated with any device. CONCLUSIONS: The carbon-lined ePTFE-encapsulated stent is a novel approach to arterial stenting. The progressive endothelialization and lack of inflammatory reaction may provide improved long-term patency. Further study of this stent-graft design is warranted. PMID- 11491263 TI - Arterial thrombus dissolution in vivo using a transducer-tipped, high-frequency ultrasound catheter and local low-dose urokinase delivery. AB - PURPOSE: To examine the hypothesis that a transducer-tipped high-frequency ultrasound drug-delivery catheter may augment the thrombolytic effects of locally delivered low-dose urokinase and result in improved recanalization rates and reduced residual thrombotic burden. METHODS: Thrombi were induced in situ bilaterally in 5- to 6-cm-long segments of the superficial femoral arteries in 9 dogs by intraluminal thermal damage and injection of thrombin. A transducer tipped high-frequency local drug-delivery catheter was applied at 1.1 MHz and 0.6 W for 60 minutes to one superficial femoral artery segment, and an identical catheter with an inactivated ultrasound transducer was used to treat the contralateral control segment. Urokinase (5000 IU/kg) was delivered bilaterally into the thrombi during the treatment interval. RESULTS: Angiography documented TIMI grade 2 or 3 flow in 9 (100%) segments in the ultrasound-treated group versus 6 (67%) of the controls (no ultrasound) (p = 0.058). Angiographically detected distal embolization was found in 2 ultrasound-treated segments compared with 5 controls (p = 0.02). Protruding or occlusive thrombi were seen angioscopically in 8 (89%) control segments but in only 1 (11%) of the ultrasound treated arteries (p < 0.001). By histopathology, 7 (78%) segments in the control group had occlusive thrombi, whereas only 3 nonocclusive thrombi were found in the ultrasound-treatment group (p < 0.001). CONCLUSIONS: Catheter-delivered high frequency ultrasound and local low-dose urokinase infusion is efficacious for the treatment of acute thrombotic occlusions as evaluated by angiography, angioscopy, and histopathology. PMID- 11491264 TI - Percutaneous techniques for managing maldeployed or migrated stents. AB - PURPOSE: To report our experience in the percutaneous management of dislocated endovascular stents. METHODS: During a 6-year period, 28 (2.7%) patients with a maldeployed or migrated endovascular stent (20 Palmaz, 5 Wallstent, 2 Memotherm, and 1 AVE) were recognized among 1021 patients undergoing noncardiac vascular stent procedures. Percutaneous stent management was performed using balloon catheters, gooseneck snares, grasping forceps, or additional stent implantation on the basis of the stent type, degree of expansion, mechanical characteristics, and location of the stent. RESULTS: Three stents that embolized into the pulmonary artery were left in situ, but percutaneous management was successful in 23 (92%) of the remaining 25 dislocated stents (12 venous and 13 arterial stents). Twelve stents were repositioned in an alternate, stable position or the primarily intended location; 4 stents were anchored by a second stent, and 7 stents were removed percutaneously. In the 2 failed cases, the stents were retrieved using a minor surgical procedure. There were 2 minor groin hematomas but no secondary complications during a median follow-up of 26.2 months (range 1 62). CONCLUSIONS: Percutaneous management of migrated or maldeployed endovascular stents is highly effective with few complications. On the basis of our findings, these techniques should be considered the therapy of choice. PMID- 11491265 TI - Pseudoaneurysm formation at the site of external iliac artery stents: percutaneous stent-graft treatment. AB - PURPOSE: To present a patient who developed an asymptomatic large iliac pseudoaneurysm complicating stent-supported iliac artery recanalization. CASE REPORT: The pseudoaneurysm was detected in an asymptomatic 69-year-old man during routine angiography 6 months after an uncomplicated procedure to implant 3 overlapping Palmaz stents in an occluded external iliac artery. There was no evidence of stent infection. During a second intervention, the pseudoaneurysm was successfully treated by percutaneous implantation of an EndoPro System I stent graft. Contrast-enhanced spiral computed tomography at 6 and 12 months confirmed the durability of aneurysm exclusion and the patency of the endoprosthesis. CONCLUSIONS: Angioplasty-induced pseudoaneurysm is rare and usually asymptomatic, but elective percutaneous stent-graft repair should be considered as the first treatment option. PMID- 11491266 TI - Intravascular ultrasound evidence for stabilization of compensatory enlargement of the femoropopliteal segment after endograft placement. AB - PURPOSE: To document whether the vasodilatory response seen at the anastomotic segment 6 months after placement of a balloon-expandable endograft in the femoropopliteal segment progresses between 6 and 24 months. METHODS: Twelve patients (9 men; median age 65 years, range 47-75) treated with an investigational polytetrafluoroethylene (PTFE) endograft for obstructive disease of the femoropopliteal segment were studied with intravascular ultrasound (IVUS) immediately after placement and at 6 months (first follow-up period) and 24 months (second follow-up period). Matched IVUS cross sections derived from the endograft and the anastomotic segment were analyzed for changes in lumen (LA), vessel (VA), and plaque areas (PLA). RESULTS: Five patients had complete IVUS surveillance at both the first (mean 8 months, range 7-9) and second (mean 25 months, range 23-26) follow-up periods; 1 patient was lost to follow-up during the second interval, and another 6 were excluded owing to graft occlusion (n = 4) or no IVUS surveillance available (n = 2) during the second follow-up period. Matched IVUS cross sections derived from the endograft showed no significant change in LA during both follow-up periods (-8% and +1%, respectively). There was no evidence for intimal hyperplasia or endograft recoil. During both follow-up periods, IVUS cross sections derived from the anastomotic segment revealed significant increases in LA (+37% and +8%, respectively) and VA (+26% and +6%, respectively) (both p < 0.05). The change in PLA during both follow-up periods was not significant (+13% and +3%, respectively). CONCLUSIONS: The PTFE endograft seems to inhibit both intimal hyperplasia and constrictive remodeling. The short term (6-month) vascular dilatory response seen at the anastomotic segment tends to stabilize at 2 years. Therefore, this endovascular anastomosis acts as an "ideal" end-to-end anastomosis. PMID- 11491267 TI - Endovascular stent-grafting in the treatment of superficial femoral artery occlusive disease. AB - PURPOSE: To explore the feasibility of using a flexible stent-graft in the treatment of superficial femoral artery occlusive lesions. METHODS: Thirty-five patients (26 men; median age 65 years, range 41-85) with long-segment occlusions of the femoropopliteal arteries were treated with the Hemobahn endovascular prosthesis. The median occlusion length was 22 cm (range 4-40). Endografting was performed in conjunction with additional recanalization procedures as needed. RESULTS: Primary recanalization and deployment of 47 Hemobahn endografts was successful in all cases; 12 patients had 2 devices implanted. The adjunctive recanalization techniques included balloon thrombectomy (n = 28), ring-stripper disobliteration (n = 7), local thrombolysis (n = 2), mechanical rotational thrombectomy (n = 2), local thromboendarterectomy and femoral bifurcation patch angioplasty (n = 14), iliac artery stenting (n = 3), and bypass grafting (n = 2). The median ankle-brachial index rose from 0.25 preoperatively to 0.87. Over an average 7-month follow-up (range 1-18), 10 (28.6%) occlusions occurred; 6 stent graft thromboses were treated successfully with thrombectomy only. The other 4 occlusions were permanent. Life-table analysis documented cumulative primary and secondary patency rates of 73.2% and 82.6%, respectively, at 1 year. CONCLUSIONS: Intraoperative use of the Hemobahn endograft is feasible and effective in restoring adequate distal perfusion in the short term, especially when combined with conventional recanalization techniques. PMID- 11491268 TI - Ischemic transverse myelopathy after endovascular repair of a thoracic aortic aneurysm. AB - PURPOSE: To report a dramatic complication after endovascular repair of a descending thoracic aortic aneurysm (TAA) and to present a classification system and possible methods to avoid spinal cord ischemia. CASE REPORT: A 48-year-old man with a descending TAA between T5 and T9 was treated with endovascular stent grafts. Fourteen hours after the operation, the patient developed partial transverse myelopathy at level T10. During emergency conversion to open surgery and implantation of a conventional tube graft, 3 intercostal arteries that had been covered by the stent-graft were revascularized. Postoperatively, the neurological deficit improved, and the patient was able to walk again. Methods to predict and possibly prevent the induction of spinal cord ischemia after endovascular repair of TAA are suggested. CONCLUSIONS: Endovascular repair of TAA may induce spinal cord ischemia; pre- and intraoperative assessment of involved intercostal arteries should be performed. PMID- 11491269 TI - Re: Intracranial vertebral artery intervention: impressive and controversial. PMID- 11491270 TI - President's year in review. PMID- 11491271 TI - On the horizon: an overview of the Student National Medical Association's plans in the upcoming year. PMID- 11491272 TI - Bacteriuria in the elderly population in a developing country. AB - Among 1470 elderly hospitalized and nonhospitalized people, 566 cases of bacteriuria were identified. There were 663 men (41.5% with bacteriuria) and 807 women (36.0% with bacteriuria). The overall prevalence of bacteriuria was 38.5%. More than 90% of the isolates were gram-negative organisms with Proteus species being the predominant pathogen among men, with 68.1% seen among inpatients. Escherichia coli was the main pathogen in women, with 62.0% recovered from inpatients. Catheterization was seen most commonly among non-hospitalized males with outflow obstruction. Catheter care in this population is often performed at home by these men who either refuse prostate surgery, are not fit for surgery, or are awaiting surgery. Polymicrobic bacteriuria was identified more frequently (approximately 60%) among the catheterized group. Of the 440 gram-negative bacilli recovered as single organisms, 352 (80.0%) were resistant to ampicillin, cephalothin, and tetracycline, whereas 229 (52.0%) were resistant to co trimoxazole. The most effective antibiotics (in increasing order of sensitivity; 80% - 100%) were augmentin, nalidixic acid, cefuroxime, gentomicin, and ciprofloxacin. No mortality due to bacteremia complicated by bacteriuria was observed during the study period. PMID- 11491273 TI - Antiretroviral adherence and use of alternative therapies among older HIV infected adults. AB - OBJECTIVE: To investigate adherence to antiretroviral therapy and use of alternative therapies among older human immunodeficiency virus (HIV)-infected adults, and to assess relationships between antiretroviral adherence and clinical outcomes. METHODS: One hundred older HIV-infected patients, aged 50 and over, treated at two large HIV clinics in Washington, DC, were enrolled. A cross sectional methodology used structured interviews to investigate antiretroviral regimens, use of alternative therapies, and demographics. Medical records provided viral load and CD4 count within 3 months of interview. RESULTS: The mean self-reported adherence was 94%, and 55 patients reported 100% adherence to antiretroviral therapy. Correlation analysis showed a significant negative correlation between adherence and viral load (r = -312, p = 0.005). There was no significant difference in adherence based on race, gender, mode of transmission, or education. Twenty-one patients (21%) reported the use of an alternative therapy, with several patients using multiple alternative therapies. There was no significant difference in adherence score (p = 0.514) or viral load (p = 0.860) based upon use of alternative therapies. CONCLUSIONS: Older HIV-infected study patients reported high levels of adherence to antiretroviral regimens, and adherence was highly correlated with HIV viral load. Use of alternative therapies did not significantly impact adherence to antiretroviral agents or viral load. High adherence among this older population may be related to older patients' familiarity with medication usage, their increasing awareness of HIV as a disease that requires optimal adherence, and educational efforts promoted by the two clinics in which they are clients. PMID- 11491274 TI - Nonfunctional parathyroid carcinoma. AB - Parathyroid carcinoma is a rare entity accounting for 0.5% to 5% of parathyroid neoplasia. Most of these malignancies present as functional hormone-producing masses with elevated serum levels of parathormone and calcium. These tumors may also be nonfunctional. Clinical detection of nonfunctioning parathyroid malignancies preoperatively is primarily based on symptoms of an expanding neck mass. This ominous complaint is typically accompanied with an advanced stage of the disease at initial diagnosis. Because there is a paucity of data in the literature regarding nonfunctioning parathyroid carcinoma, prognosis can not be readily assessed. In both functional and nonfunctional parathyroid carcinoma, early surgery has proven to be the only curative treatment approach whereas both chemotherapy and radiation therapy fail to produce systemic or regional benefit when used alone. Hence, parathyroid cancer should be considered in every patient evaluated for a neck mass regardless of the blood calcium and blood parathormone level. PMID- 11491275 TI - Cyclic gmp is a measure of physiologic stress. AB - This study was undertaken to determine whether patients who were critically ill evidenced elevated levels of blood cyclic guanosine monophosphate (cGMP). Cyclic guanosine monophosphate levels correlated with severity of illness as measured by the APACHE II severity of illness scoring system (p < 0.01). Cyclic guanosine monophosphate also correlated with the level of carboxyhemoglobin (HbCO) (p < 0.001). The correlation between cGMP and creatinine was p < 0.0001. Patients with end-stage disease (renal or liver) tended to have elevated levels of cGMP (p < 0.0001). We conclude that the induction of these two molecules may be linked in patients with increasing severity of illness. PMID- 11491276 TI - Health disparity in black women: lack of pharmaceutical advertising in black vs. white-oriented magazines. AB - CONTEXT: Racial disparities in health care between black women and white women may be attributed in part to socioeconomic status and lack of insurance, but also may be due to lack of the dissemination of health information in black communities via black popular magazines. OBJECTIVE: Comparison of the number and type of pharmaceutical advertisements between black-oriented magazines and white oriented magazines. DESIGN: Descriptive study. SETTING: Morehouse School of Medicine. MAIN OUTCOME MEASURES: Recording of the type and number of over-the counter and prescription drug advertisements. RESULTS: Five black-oriented magazines (Black Woman, Black Elegance, Essence, Ebony, and Upscale) and 5 white oriented magazines (Family Circle, Working Mother, Good Housekeeping, Ladies' Home Journal, Women's Day) were evaluated for 3 months from June-August, 2000. White-oriented magazines had four to eight times more pharmaceutical ads than black-oriented magazines. Types of medications advertised in the white-oriented magazines and not the black-oriented magazines were, for example, cholinesterase inhibitors, calcium supplements, COX II-inhibitors, intranasal steroids, anorexiants, proton pump inhibitors, and smoking deterrent agents. Conversely, medications advertised in the black-oriented magazines and not the white-oriented magazines were antiviral agents and oral contraceptives. Pharmaceutical companies gave several reasons for the disparity, including the explanation that their particular company was advertising about HIV in the black community. CONCLUSIONS: A barrier to equitable health care for black women may be a low prioritization for health prevention and health management. This low prioritization or disinterest may be a reflection of the black magazine that she is reading due to the lack of pharmaceutical advertisements in black-oriented magazines. The result of this disinterest of black females may be seen in the increased morbidity and mortality for selected diseases. PMID- 11491278 TI - Streptococcus milleri: a cause of pyogenic liver abscess. AB - We report a case of Streptococcus milleri in a patient with a history of hypertension, alcohol abuse, and tobacco smoking. S. milleri is associated with liver abscesses much more frequently than other streptococci. PMID- 11491277 TI - Strenuous physical activity and breast cancer risk in African-American women. AB - OBJECTIVES: Some studies of white women suggest that exercise reduces the incidence of breast cancer. There are no data on black women. We assessed the relationship between strenuous physical activity and prevalent breast cancer among participants in the Black Women's Health Study. METHODS: Data on strenuous recreational physical activity at various ages and other factors were collected in 1995 by mail questionnaire from 64,524 United States black women aged 21 to 69 years. The 704 women who reported breast cancer (cases) were matched on age and on menopausal status at the time of the breast cancer diagnosis with 1408 women who did not report breast cancer (controls). Odds ratios for levels of physical activity at various ages were derived from conditional logistic regression with control for potential confounding factors. RESULTS: Odds ratios for > or =7 h per week relative to < 1 were significantly reduced for strenuous activity at age 21 for breast cancer overall and premenopausal breast cancer, at age 30 for breast cancer overall, and at age 40 for postmenopausal breast cancer. There was no evidence of a reduction associated with exercise in high school. CONCLUSIONS: The findings of the present study suggest that strenuous physical activity in early adulthood is associated with a reduced risk of breast cancer in African-American women. PMID- 11491279 TI - Radiation dose monitoring in a breast cancer patient with a pacemaker: a case report. AB - A pacemaker-bearing patient with left-sided breast cancer was treated with adjuvant external beam radiation therapy to the intact breast. She was treated via tangential fields and a single anterior supraclavicular field using 6-MV x rays. The pacemaker, originally in the treatment field, was removed and a new one placed 4 cm outside the radiation field prior to treatment. Silicon diode chamber Keithley-Farmer type 0.6 cc ionization chamber, and lithium fluoride (LiF) (TLD) chips were used to measure, in vivo, the dose to the pacemaker. From all the fields treated, total dose to the pacemaker was 164 cGy by diode measurements, 182 cGy by ionization chamber measurements, and 171 cGy by TLD measurements. The pacemaker functioned normally throughout the course of treatment. PMID- 11491280 TI - Thoracic aortic dissection in a patient with autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease is one of the most common hereditary diseases, and frequently has well defined extrarenal manifestations. Very few cases of aortic aneurysms associated with this disorder are described in literature. We report a 42-year-old male with autosomal dominant polycystic kidney disease presenting with dissecting aneurysm of the thoracic aorta. PMID- 11491281 TI - Therapeutic options: hormone replacement therapy-soy therapy. PMID- 11491282 TI - Comparison of ARM and HEAT protein repeats. AB - ARM and HEAT motifs are tandemly repeated sequences of approximately 50 amino acid residues that occur in a wide variety of eukaryotic proteins. An exhaustive search of sequence databases detected new family members and revealed that at least 1 in 500 eukaryotic protein sequences contain such repeats. It also rendered the similarity between ARM and HEAT repeats, believed to be evolutionarily related, readily apparent. All the proteins identified in the database searches could be clustered by sequence similarity into four groups: canonical ARM-repeat proteins and three groups of the more divergent HEAT-repeat proteins. This allowed us to build improved sequence profiles for the automatic detection of repeat motifs. Inspection of these profiles indicated that the individual repeat motifs of all four classes share a common set of seven highly conserved hydrophobic residues, which in proteins of known three-dimensional structure are buried within or between repeats. However, the motifs differ at several specific residue positions, suggesting important structural or functional differences among the classes. Our results illustrate that ARM and HEAT-repeat proteins, while having a common phylogenetic origin, have since diverged significantly. We discuss evolutionary scenarios that could account for the great diversity of repeats observed. PMID- 11491283 TI - The RegB/RegA two-component regulatory system controls synthesis of photosynthesis and respiratory electron transfer components in Rhodobacter capsulatus. AB - Recently, we demonstrated that the RegB/RegA two-component regulatory system from Rhodobacter capsulatus functions as a global regulator of metabolic processes that either generate or consume reducing equivalents. For example, the RegB/RegA system controls expression of such energy generating processes as photosynthesis and hydrogen utilization. In addition, RegB/RegA also control nitrogen and carbon fixation pathways that utilize reducing equivalents. Here, we use a combination of DNase I protection and plasmid-based reporter expression studies to demonstrate that RegA directly controls synthesis of cytochrome cbb3 and ubiquinol oxidases that function as terminal electron acceptors in a branched respiratory chain. We also demonstrate that RegA controls expression of cytochromes c2, c(y) and the cytochrome bc1 complex that are involved in both photosynthetic and respiratory electron transfer events. These data provide evidence that the RegB/RegA two-component system has a major role in controlling the synthesis of numerous processes that affect reducing equivalents in Rhodobacter capsulatus. PMID- 11491284 TI - The RNA i-motif. AB - Oligodeoxynucleotides with stretches of cytidine residues associate into a four stranded structure, the i-motif, in which two head-to-tail, intercalated, parallel-stranded duplexes are held together by hemiprotonated C.C+ pairs. We have investigated the possibility of forming an i-motif structure with C-rich ribonucleic acids. The four C-rich RNAs studied, r(UC5), r(C5), r(C5U) and r(UC3), associate into multiple intercalated structures at acidic pH. r(UC5) forms two i-motif structures that differ by their intercalation topologies. We report on a structural study of the main form and we analyze the small conformational differences found by comparison with the DNA i-motif. The stacking topology of the main structure avoids one of the six 2'-OH/2'-OH repulsive contacts expected in a fully intercalated structure. The C3'-endo pucker of the RNA sugars and the orientation of the intercalated C.C+ pairs result in a modest widening of the narrow grooves at the steps where the hydroxyl groups are in close contact. The free energy of the RNA i-motif, on average -4 kJ mol(-1) per C.C+ pair, is half of the value found in DNA i-motif structures. PMID- 11491285 TI - The N-terminal domain of Homer/Vesl is a new class II EVH1 domain. AB - Cellular activities controlled by signal transduction processes such as cell motility and cell growth depend on the tightly regulated assembly of multiprotein complexes. Adapter proteins that specifically interact with their target proteins are key components required for the formation of these assemblies. Ena/VASP homology 1 (EVH1) domains are small constituents of large modular proteins involved in microfilament assembly that specifically recognize proline-rich regions. EVH1 domain-containing proteins are present in neuronal cells, like the Homer/Vesl protein family that is involved in memory-generating processes. Here, we describe the crystal structure of the murine EVH1 domain of Vesl 2 at 2.2 A resolution. The small globular protein consists of a seven-stranded antiparallel beta-barrel with a C-terminal alpha-helix packing alongside the barrel. A shallow groove running parallel with beta-strand VI forms an extended peptide-binding site. Using peptide library screenings, we present data that demonstrate the high affinity of the Vesl 2 EVH1 domain towards peptide sequences containing a proline rich core sequence (PPSPF) that requires additional charged amino acid residues on either side for specific binding. Our functional data, substantiated by structural data, demonstrate that the ligand-binding of the Vesl EVH1 domain differs from the interaction characteristics of the previously examined EVH1 domains of the Evl/Mena proteins. Analogous to the Src homology 3 (SH3) domains that bind their cognate ligands in two distinct directions, we therefore propose the existence of two distinct classes of EVH1 domains. PMID- 11491286 TI - A study of the structure-activity relationship for diazaborine inhibition of Escherichia coli enoyl-ACP reductase. AB - Enoyl acyl carrier protein (ACP) reductase catalyses the last reductive step of fatty acid biosynthesis, reducing the enoyl group of a growing fatty acid chain attached to ACP to its acyl product using NAD(P)H as the cofactor. This enzyme is the target for the diazaborine class of antibacterial agents, the biocide triclosan, and one of the targets for the front-line anti-tuberculosis drug isoniazid. The structures of complexes of Escherichia coli enoyl-ACP reductase (ENR) from crystals grown in the presence of NAD+ and a family of diazaborine compounds have been determined. Analysis of the structures has revealed that a mobile loop in the structure of the binary complex with NAD+ becomes ordered on binding diazaborine/NAD+ but displays a different conformation in the two subunits of the asymmetric unit. The work presented here reveals how, for one of the ordered conformations adopted by the mobile loop, the mode of diazaborine binding correlates well with the activity profiles of the diazaborine family. Additionally, diazaborine binding provides insights into the pocket on the enzyme surface occupied by the growing fatty acid chain. PMID- 11491287 TI - Binding site differences revealed by crystal structures of Plasmodium falciparum and bovine acyl-CoA binding protein. AB - Acyl-CoA binding protein (ACBP) maintains a pool of fatty acyl-CoA molecules in the cell and plays a role in fatty acid metabolism. The biochemical properties of Plasmodium falciparum ACBP are described together with the 2.0 A resolution crystal structures of a P. falciparum ACBP-acyl-CoA complex and of bovine ACBP in two crystal forms. Overall, the bovine ACBP crystal structures are similar to the NMR structures published previously; however, the bovine and parasite ACBP structures are less similar. The parasite ACBP is shown to have a different ligand-binding pocket, leading to an acyl-CoA binding specificity different from that of bovine ACBP. Several non-conservative differences in residues that interact with the ligand were identified between the mammalian and parasite ACBPs. These, together with measured binding-specificity differences, suggest that there is a potential for the design of molecules that might selectively block the acyl-CoA binding site. PMID- 11491288 TI - The antiterminator NusB enhances termination at a sub-optimal Rho site. AB - Interactions between the antiterminator NusB and boxA elements in the nut sites are necessary to ensure lambda N-mediated processive antitermination. Similarly, in the bacterial cell, interactions between NusB and boxA elements help RNA polymerase to counteract polarity during transcription of rrn operons. We analyzed the effects of NusB on intragenic termination at the level of two tandem terminators located in the hisG cistron, GTTE1 and GTTE2. Unexpectedly, we found that NusB enhances transcription termination at the sub-optimal Rho site GTTE1. Moreover, site-directed mutagenesis of a boxA homolog located within GTTE1 and the masking of this element by translating ribosomes demonstrated that the recruitment of NusB in the termination complex is mediated by a boxA element. The mutated boxA also abolishes the formation of a NusB-dependent complex on GTTE1 RNA. On the whole, results provide evidence that interactions between NusB and boxA can enhance Rho-dependent termination. PMID- 11491289 TI - Weak protein-protein interactions in lectins: the crystal structure of a vegetative lectin from the legume Dolichos biflorus. AB - The legume lectins are widely used as a model system for studying protein carbohydrate and protein-protein interactions. They exhibit a fascinating quaternary structure variation, which becomes important when they interact with multivalent glycoconjugates, for instance those on cell surfaces. Recently, it has become clear that certain lectins form weakly associated oligomers. This phenomenon may play a role in the regulation of receptor crosslinking and subsequent signal transduction. The crystal structure of DB58, a dimeric lectin from the legume Dolichos biflorus reveals a separate dimer of a previously unobserved type, in addition to a tetramer consisting of two such dimers. This tetramer resembles that formed by DBL, the seed lectin from the same plant. A single amino acid substitution in DB58 affects the conformation and flexibility of a loop in the canonical dimer interface. This disrupts the formation of a stable DBL-like tetramer in solution, but does not prohibit its formation in suitable conditions, which greatly increases the possibilities for the cross linking of multivalent ligands. The non-canonical DB58 dimer has a buried symmetrical alpha helix, which can be present in the crystal in either of two antiparallel orientations. Two existing structures and datasets for lectins with similar quaternary structures were reconsidered. A central alpha helix could be observed in the soybean lectin, but not in the leucoagglutinating lectin from Phaseolus vulgaris. The relative position and orientation of the carbohydrate binding sites in the DB58 dimer may affect its ability to crosslink mulitivalent ligands, compared to the other legume lectin dimers. PMID- 11491290 TI - The structure of Escherichia coli nitroreductase complexed with nicotinic acid: three crystal forms at 1.7 A, 1.8 A and 2.4 A resolution. AB - Escherichia coli nitroreductase is a flavoprotein that reduces a variety of quinone and nitroaromatic substrates. Its ability to convert relatively non-toxic prodrugs such as CB1954 (5-[aziridin-1-yl]-2,4-dinitrobenzamide) into highly cytotoxic derivatives has led to interest in its potential for cancer gene therapy. We have determined the structure of the enzyme bound to a substrate analogue, nicotinic acid, from three crystal forms at resolutions of 1.7 A, 1.8 A and 2.4 A, representing ten non-crystallographically related monomers. The enzyme is dimeric, and has a large hydrophobic core; each half of the molecule consists of a five-stranded beta-sheet surrounded by alpha-helices. Helices F and F protrude from the core region of each monomer. There is an extensive dimer interface, and the 15 C-terminal residues extend around the opposing monomer, contributing the fifth beta-strand. The active sites lie on opposite sides of the molecule, in solvent-exposed clefts at the dimer interface. The FMN forms hydrogen bonds to one monomer and hydrophobic contacts to both; its si face is buried. The nicotinic acid stacks between the re face of the FMN and Phe124 in helix F, with only one hydrogen bond to the protein. If the nicotinamide ring of the coenzyme NAD(P)H were in the same position as that of the nicotinic acid ligand, its C4 atom would be optimally positioned for direct hydride transfer to flavin N5. Comparison of the structure with unliganded flavin reductase and NTR suggests reduced mobility of helices E and F upon ligand binding. Analysis of the structure explains the broad substrate specificity of the enzyme, and provides the basis for rational design of novel prodrugs and for site-directed mutagenesis for improved enzyme activity. PMID- 11491291 TI - The crystal structure of Bacillus subtilis lipase: a minimal alpha/beta hydrolase fold enzyme. AB - The X-ray structure of the lipase LipA from Bacillus subtilis has been determined at 1.5 A resolution. It is the first structure of a member of homology family 1.4 of bacterial lipases. The lipase shows a compact minimal alpha/beta hydrolase fold with a six-stranded parallel beta-sheet flanked by five alpha-helices, two on one side of the sheet and three on the other side. The catalytic triad residues, Ser77, Asp133 and His156, and the residues forming the oxyanion hole (backbone amide groups of Ile12 and Met78) are in positions very similar to those of other lipases of known structure. However, no lid domain is present and the active-site nucleophile Ser77 is solvent-exposed. A model of substrate binding is proposed on the basis of a comparison with other lipases with a covalently bound tetrahedral intermediate mimic. It explains the preference of the enzyme for substrates with C8 fatty acid chains. PMID- 11491292 TI - Direct observation of three conformations of MutS protein regulated by adenine nucleotides. AB - Mismatched base-pairs, which are caused by either DNA replication errors, DNA damage or genetic recombination, are repaired by the mismatch-repair system. The MutS protein, a component of the mismatch-repair system, recognizes mismatched base-pairs in DNA, and its DNA-binding activity is affected by ATP and ADP. Here, we show that the MutS protein from Thermus thermophilus HB8 can have three different conformations in solution, based on direct observations made by small angle X-ray scattering. The conformation of MutS in solution is drastically influenced by the presence of ADP and ATP; the ATP-bound form has the most compact conformation, the ADP-bound form the most stretched, and the nucleotide free form has a conformation intermediate between the two. Based on these findings, we conclude that the DNA-binding activity of MutS may depend on conformational changes triggered by both the binding and hydrolysis of ATP. PMID- 11491293 TI - Mechanism of hydrolysis of phosphate esters by the dimetal center of 5' nucleotidase based on crystal structures. AB - 5'-Nucleotidase belongs to a large superfamily of distantly related dinuclear metallophosphatases including the Ser/Thr protein phosphatases and purple acid phosphatases. The protein undergoes a 96 degrees domain rotation between an open (inactive) and a closed (active) enzyme form. Complex structures of the closed form with the products adenosine and phosphate, and with the substrate analogue inhibitor alpha,beta-methylene ADP, have been determined at 2.1 A and 1.85 A resolution, respectively. In addition, a complex of the open form of 5' nucleotidase with ATP was analyzed at a resolution of 1.7 A. These structures show that the adenosine group binds to a specific binding pocket of the C terminal domain. The adenine ring is stacked between Phe429 and Phe498. The N terminal domain provides the ligands to the dimetal cluster and the conserved His117, which together form the catalytic core structure. However, the three C terminal arginine residues 375, 379 and 410, which are involved in substrate binding, may also play a role in transition-state stabilization. The beta phosphate group of the inhibitor is terminally coordinated to the site 2 metal ion. The site 1 metal ion coordinates a water molecule which is in an ideal position for a nucleophilic attack on the phosphorus atom, assuming an in-line mechanism of phosphoryl transfer. Another water molecule bridges the two metal ions. PMID- 11491294 TI - E. coli 5'-nucleotidase undergoes a hinge-bending domain rotation resembling a ball-and-socket motion. AB - Structures of nine independent conformers of E. coli 5'-nucleotidase (5'-NT) have been analyzed using four different crystal forms. These data show that the two domain protein undergoes an unusual 96 degrees hinge-bending domain rotation. Structures of the open and closed forms with substrates and inhibitors reveal that the substrate moves by approximately 25 A with the large domain rotation into the catalytic site. The domain motions derived from a comparison of the nine conformations agree well with motions obtained from a normal mode analysis in that all independent domain rotations are around axes that are roughly located in the plane which includes the domain centers and the hinge. Two residues, Lys355 and Gly356, form the core of the hinge region and undergo a large change of the main-chain torsion angles. The hinge-bending movement observed for 5' nucleotidase differs markedly from a classical hinge-bending closure motion which involves an opening of the substrate or ligand-binding cleft between two domains. In contrast, the movement observed in 5'-nucleotidase resembles that of a ball and-socket joint. The smaller C-terminal domain rotates approximately around its center such that the residues at the domain interface move in a sliding motion along the interface. Few direct interdomain contacts and a layer of water molecules between the two domains facilitate the sliding motion. PMID- 11491295 TI - Solution structures of C-1027 apoprotein and its complex with the aromatized chromophore. AB - C-1027 is one of the most potent antitumor antibiotic chromoproteins, and is a 1:1 complex of an enediyne chromophore having DNA-cleaving ability and a carrier apoprotein. The three-dimensional solution structures of the 110 residue (10.5 kDa) C-1027 apoprotein and its complex with the aromatized chromophore have been determined separately by homonuclear two-dimensional nuclear magnetic resonance methods. The apoprotein is mainly composed of three antiparallel beta-sheets: four-stranded beta-sheet (43-45, 52-54; 30-38; 92-94; 104-106), three-stranded beta-sheet (4-6; 17-22; 61-66), and two-stranded beta-sheet (70-72; 83-85). The overall structure of the apoprotein is very similar to those of other chromoprotein apoproteins, such as neocarzinostatin and kedarcidin. A hydrophobic pocket with approximate dimensions of 14 A x 12 A x 8 A is formed by the four stranded beta-sheet and the three loops (39-42; 75-79; 97-100). The holoprotein (complex form with the aromatized chromophore) structure reveals that the aromatized chromophore is bound to the hydrophobic pocket found in the apoprotein. The benzodihydropentalene core of the chromophore is located in the center of the pocket and other substituents (beta-tyrosine, benzoxazine, and aminosugar moieties) are arranged around the core. Major binding interactions between the apoprotein and the chromophore are likely the hydrophobic contacts between the core of the chromophore and the hydrophobic side-chains of the pocket forming residues, which is supplemented by salt bridges and/or hydrogen bonds. Based on the holoprotein structure, we propose possible mechanisms for the stabilization and the release of chromophore by the apoprotein. PMID- 11491296 TI - Role of native topology investigated by multiple unfolding simulations of four SH3 domains. AB - The relative importance of amino acid sequence and native topology in the unfolding process of two SH3 domains and two circular permutants was investigated by 120 molecular dynamics runs at 375 K for a total simulation time of 0.72 micros. The alpha-spectrin (aSH3) and src SH3 (sSH3) domains, which have the same topology and a sequence identity of only 34%, show similar unfolding pathways. The disappearance of the three-stranded antiparallel beta-sheet is the last unfolding event, in agreement with a large repertoire of kinetic data derived from point mutations as well as glycine insertions and disulfide crosslinks. Two alternative routes of beta-sheet unfolding have emerged from the analysis of the trajectories. One is statistically preferred in aSH3 (n-src loop breaks before distal hair-pin) and the inverse in sSH3. An elongation of the beta2-beta3 hairpin was observed during the unfolding of sSH3 at 375 K and in 300 K simulations started from the putative transition state of sSH3 in accord with unusual kinetic data for point mutations at the n-src loop. The change of connectivity in the permutants influenced the sequence of unfolding events mainly at the permutation site. Regions where the connectivity remained unaffected showed the same chronology of contact disappearance. Taken together with previous folding simulations of two designed three-stranded antiparallel beta-sheet peptides, these results indicate that, at least for small beta-sheet proteins, the folding mechanism is primarily defined by the native state topology, whilst specific interactions determine the statistically predominant folding route. PMID- 11491297 TI - Appropriate initiation of the strand exchange reaction promoted by RecA protein requires ATP hydrolysis. AB - The DNA-dependent ATPase activity of the Escherichia coli RecA protein has been recognized for more than two decades. Yet, the role of ATP hydrolysis in the RecA promoted strand exchange reaction remains unclear. Here, we demonstrate that ATP hydrolysis is required as part of a proofreading process during homology recognition. It enables the RecA-ssDNA complex, after determining that the strand exchanged duplex is mismatched, to dissociate from the synaptic complex, which allows it to re-initiate the search for a "true" homologous region. Furthermore, the results suggest that when non-homologous sequences are present at the proximal end, ATP hydrolysis is required to allow ssDNA-RecA to reinitiate the strand exchange from an internal homologous region. PMID- 11491298 TI - Essential dynamics of reversible peptide folding: memory-free conformational dynamics governed by internal hydrogen bonds. AB - A principal component analysis has been applied on equilibrium simulations of a beta-heptapeptide that shows reversible folding in a methanol solution. The analysis shows that the configurational space contains only three dense sub states. These states of relatively low free energy correspond to the "native" left-handed helix, a partly helical intermediate, and a hairpin-like structure. The collection of unfolded conformations form a relatively diffuse cloud with little substructure. Internal hydrogen-bonding energies were found to correlate well with the degree of folding. The native helical structure folds from the N terminus; the transition from the major folding intermediate to the native helical structure involves the formation of the two most C-terminal backbone hydrogen bonds. A four-state Markov model was found to describe transition frequencies between the conformational states within error limits, indicating that memory-effects are negligible beyond the nanosecond time-scale. The dominant native state fluctuations were found to be very similar to unfolding motions, suggesting that unfolding pathways can be inferred from fluctuations in the native state. The low-dimensional essential subspace, describing 69% of the collective atomic fluctuations, was found to converge at time-scales of the order of one nanosecond at all temperatures investigated, whereas folding/unfolding takes place at significantly longer time-scales, even above the melting temperature. PMID- 11491299 TI - On the mechanism of biological methane formation: structural evidence for conformational changes in methyl-coenzyme M reductase upon substrate binding. AB - Methyl-coenzyme M reductase (MCR) catalyzes the final reaction of the energy conserving pathway of methanogenic archaea in which methylcoenzyme M and coenzyme B are converted to methane and the heterodisulfide CoM-S-S-CoB. It operates under strictly anaerobic conditions and contains the nickel porphinoid F430 which is present in the nickel (I) oxidation state in the active enzyme. The known crystal structures of the inactive nickel (II) enzyme in complex with coenzyme M and coenzyme B (MCR-ox1-silent) and in complex with the heterodisulfide CoM-S-S-CoB (MCR-silent) were now refined at 1.16 A and 1.8 A resolution, respectively. The atomic resolution structure of MCR-ox1-silent describes the exact geometry of the cofactor F430, of the active site residues and of the modified amino acid residues. Moreover, the observation of 18 Mg2+ and 9 Na+ ions at the protein surface of the 300 kDa enzyme specifies typical constituents of binding sites for either ion. The MCR-silent and MCR-ox1-silent structures differed in the occupancy of bound water molecules near the active site indicating that a water chain is involved in the replenishment of the active site with water molecules. The structure of the novel enzyme state MCR-red1-silent at 1.8 A resolution revealed an active site only partially occupied by coenzyme M and coenzyme B. Increased flexibility and distinct alternate conformations were observed near the active site and the substrate channel. The electron density of the MCR-red1 silent state aerobically co-crystallized with coenzyme M displayed a fully occupied coenzyme M-binding site with no alternate conformations. Therefore, the structure was very similar to the MCR-ox1-silent state. As a consequence, the binding of coenzyme M induced specific conformational changes that postulate a molecular mechanism by which the enzyme ensures that methylcoenzyme M enters the substrate channel prior to coenzyme B as required by the active-site geometry. The three different enzymatically inactive enzyme states are discussed with respect to their enzymatically active precursors and with respect to the catalytic mechanism. PMID- 11491300 TI - Effects of cobalt hexammine on folding and self-cleavage of the Neurospora VS ribozyme. AB - We have investigated the effects of Co(NH3)6(3+), an analog of hexahydrated Mg2+, on folding and catalysis of the Neurospora VS ribozyme. Most of the metal ion induced changes detected by chemical modification structure probing in either metal ion are similar, but occur at approximately 33-fold lower concentrations of Co(NH3)6(3+) than Mg2+. However, Co(NH3)6(3+) is not as effective at inducing two functionally important structural changes: stabilizing the pseudoknot interaction between loops I and V, and rearranging the secondary structure of helix Ib. Comparison of the folding of the precursor and the downstream cleavage product, which lacks helix Ia, shows that helix Ia inhibits stable pseudoknot formation and rearrangement of helix Ib. The VS ribozyme does not self-cleave with Co(NH3)6(3+) as the sole polyvalent cation; however, mixed-metal kinetic experiments show that Co(NH3)6(3+) does not inhibit Mg2+-induced self-cleavage. In contrast, at sub-saturating concentrations of Mg2+, Co(NH3)6(3+) increases the rate of Mg2+-induced self-cleavage, indicating that Co(NH3)6(3+) contributes to the functionally relevant folding of the VS ribozyme. PMID- 11491301 TI - Role of counterion condensation in folding of the Tetrahymena ribozyme. II. Counterion-dependence of folding kinetics. AB - Condensed counterions contribute to the stability of compact structures in RNA, largely by reducing electrostatic repulsion among phosphate groups. Varieties of cations induce a collapsed state in the Tetrahymena ribozyme that is readily transformed to the catalytically active structure in the presence of Mg2+. Native gel electrophoresis was used to compare the effects of the valence and size of the counterion on the kinetics of this transition. The rate of folding was found to decrease with the charge of the counterion. Transitions in monovalent ions occur 20- to 40-fold faster than transitions induced by multivalent metal ions. These results suggest that multivalent cations yield stable compact structures, which are slower to reorganize to the native conformation than those induced by monovalent ions. The folding kinetics are 12-fold faster in the presence of spermidine3+ than [Co(NH3)6]3+, consistent with less effective stabilization of long-range RNA interactions by polyamines. Under most conditions, the observed folding rate decreases with increasing counterion concentration. In saturating amounts of counterion, folding is accelerated by addition of urea. These observations indicate that reorganization of compact intermediates involves partial unfolding of the RNA. We find that folding of the ribozyme is most efficient in a mixture of monovalent salt and Mg2+. This is attributed to competition among counterions for binding to the RNA. The counterion dependence of the folding kinetics is discussed in terms of the ability of condensed ions to stabilize compact structures in RNA. PMID- 11491302 TI - FokI requires two specific DNA sites for cleavage. AB - FokI is a bipartite restriction endonuclease that recognizes a non-palindromic DNA sequence, and then makes double-stranded cuts outside of that sequence to leave a 5' overhang. Earlier kinetic and crystallographic studies suggested that FokI might function as a dimer. Here, we show, using dynamic light-scattering, gel-filtration and analytical ultracentrifugation, that FokI dimerizes only in the presence of divalent metal ions. Furthermore, analysis of the DNA-bound complex reveals that two copies of the recognition sequence are incorporated into the dimeric complex and that formation of this complex is essential for full activation of cleavage. These results have broad implications for the mechanism by which monomeric type II endonucleases achieve high fidelity. PMID- 11491303 TI - Structural requirements for cooperative binding of HMG1 to DNA minicircles. AB - DNA minicircles, where the length of DNA is below the persistence length, are highly effective, preferred, ligands for HMG-box proteins. The proteins bind to them "structure-specifically" with affinities in the nanomolar range, presumably to an exposed widened minor groove. To understand better the basis of this preference, we have studied the binding of HMG1 (which has two tandem HMG boxes linked by a basic extension to a long acidic tail) and Drosophila HMG-D (one HMG box linked by a basic region to a short and less acidic tail), and their HMG-box domains, to 88 bp and 75 bp DNA minicircles. In some cases we see cooperative binding of two molecules to the circles. The requirements for strong cooperativity are two HMG boxes and the basic extension; the latter also appears to stabilize and constrain the complex, preventing binding of further protein molecules. HMG-D, with a single HMG box, does not bind cooperatively. In the case of HMG1, the acidic tail is not required for cooperativity and does not affect binding significantly, in contrast to a much greater effect with linear DNA, or even four-way junctions (another distorted DNA substrate). Such effects could be relevant in the hierarchy of binding of HMG-box proteins to DNA distortions in vivo, where both single-box and two-box proteins might co-exist, with or without basic extensions and acidic tails. PMID- 11491304 TI - Covalent joining of the subunits of a homodimeric type II restriction endonuclease: single-chain PvuII endonuclease. AB - The PvuII restriction endonuclease has been converted from its natural homodimeric form into a single polypeptide chain by tandemly linking the two subunits through a short peptide linker. The arrangement of the single-chain PvuII (sc PvuII) is (2-157)-GlySerGlyGly-(2-157), where (2-157) represents the amino acid residues of the enzyme subunit and GlySerGlyGly is the peptide linker. By introducing the corresponding tandem gene into Escherichia coli, PvuII endonuclease activity could be detected in functional in vivo assays. The sc enzyme was expressed at high level as a soluble protein. The purified enzyme was shown to have the molecular mass expected for the designed sc protein. Based on the DNA cleavage patterns obtained with different substrates, the cleavage specificity of the sc PvuII is indistinguishable from that of the wild-type (wt) enzyme. The sc enzyme binds specifically to the cognate DNA site under non catalytic conditions, in the presence of Ca2+, with the expected 1:1 stoichiometry. Under standard catalytic conditions, the sc enzyme cleaves simultaneously the two DNA strands in a concerted manner. Steady-state kinetic parameters of DNA cleavage by the sc and wt PvuII showed that the sc enzyme is a potent, but somewhat less efficient catalyst; the k(cat)/K(M) values are 1.11 x 10(9) and 3.50 x 10(9) min(-1) M(-1) for the sc and wt enzyme, respectively. The activity decrease is due to the lower turnover number and to the lower substrate affinity. The sc arrangement provides a facile route to obtain asymmetrically modified heterodimeric enzymes. PMID- 11491305 TI - Computer-assisted identification of cell cycle-related genes: new targets for E2F transcription factors. AB - The processes that take place during development and differentiation are directed through coordinated regulation of expression of a large number of genes. One such gene regulatory network provides cell cycle control in eukaryotic organisms. In this work, we have studied the structural features of the 5' regulatory regions of cell cycle-related genes. We developed a new method for identifying composite substructures (modules) in regulatory regions of genes consisting of a binding site for a key transcription factor and additional contextual motifs: potential targets for other transcription factors that may synergistically regulate gene transcription. Applying this method to cell cycle-related promoters, we created a program for context-specific identification of binding sites for transcription factors of the E2F family which are key regulators of the cell cycle. We found that E2F composite modules are found at a high frequency and in close proximity to the start of transcription in cell cycle-related promoters in comparison with other promoters. Using this information, we then searched for E2F sites in genomic sequences with the goal of identifying new genes which play important roles in controlling cell proliferation, differentiation and apoptosis. Using a chromatin immunoprecipitation assay, we then experimentally verified the binding of E2F in vivo to the promoters predicted by the computer-assisted methods. Our identification of new E2F target genes provides new insight into gene regulatory networks and provides a framework for continued analysis of the role of contextual promoter features in transcriptional regulation. The tools described are available at http://compel.bionet.nsc.ru/FunSite/SiteScan.html. PMID- 11491306 TI - Prenatal diagnosis of familial hypercholesterolemia: importance of DNA analysis in the high-risk South African population. AB - In this report on the outcome of the first prenatal diagnosis performed for familial hypercholesterolemia (FH) in a South African family, we aim to demonstrate the value of a population-directed screening strategy to identify FH patients in populations with an enrichment for certain low-density lipoprotein receptor (LDLR) gene mutations. Prenatal diagnosis was offered to an Afrikaner couple, both partners heterozygous for the FH mutation D206E, whose first child was diagnosed with heterozygous FH and the second with homozygous FH. Genomic DNA isolated from parental peripheral blood and subsequently amniotic fluid was amplified by the polymerase chain reaction (PCR) and subjected to mutation analysis. Heterozygosity for mutation D206E was confirmed in both parents, whilst this mutation was not detected in DNA directly amplified from amniotic fluid. To exclude the possibility of a false-negative result due to the limited number of cells in the uncultured amniotic fluid sample, cells were also cultured in vitro, and the DNA extracted and subjected to a second round of analysis. This confirmed the absence of mutation D206E in the fetus. This case illustrates the application of a DNA-based mutation detection technique as a simple and rapid diagnostic aid that can be carried out at a relatively early gestational stage. Prenatal diagnosis of FH, aimed at the detection of homozygous cases, is particularly feasible in populations and families with molecularly defined LDLR gene mutations. PMID- 11491307 TI - One novel and two recurrent missense DKC1 mutations in patients with dyskeratosis congenita (DKC). AB - X-linked dyskeratosis congenita (DKC) is a progressive multisystem disorder most severely affecting tissues with a high cellular turnover such as skin, mucous membranes, and blood. Most patients die of bone marrow failure, although the chances of succumbing to various types of cancer and pulmonary disease are also high. DKC is caused predominantly by missense mutations in the DKC1 gene linked to Xq28. Some of the clinical features are reminiscent of premature ageing and this agrees with recent indications that DKC could be a telomere maintenance disorder. There is considerable variability in the type, severity, and age at onset of the various anomalies. Recognition of this has increased with the finding that patients with Hoyeraal-Hreidarsson syndrome (HHS) who exhibit severe neurological problems in addition to early-onset pancytopenia, also bear mutations in the DKC1 gene. For these reasons, and compounded by the range of mutations, phenotype-genotype correlations and accurate assessments of prognosis have not been possible. To complement the present data, we here report on three new cases of DKC and their mutations. One is a novel mutation in the exon 3 (K43E). The other two represent a frequently recurring mutation in exon 11 (A353V) and a less frequently recurring mutation in the exon 3 (T49M). PMID- 11491308 TI - Combined trisomy 9P and Shprintzen syndrome resulting from a paternal t(9;22). AB - The authors report on a female infant with partial trisomy 9 (pter-->q12) together with partial monosomy 22 (pter-->q11.23) that included DiGeorge critical region (DGCR), as a result of adjacent-2 disjunction. In addition to the clinical features characteristic of trisomy 9p syndrome, the patient had Truncus arteriosus type A2, bilateral hydronephrosis, palatal anomaly, retrognathia, and laryngeal hypotonia, which are likely to be attributed to 22q11.2 deletion. This patient appears to be the first reported case with such unbalanced translocation resulting from a paternal reciprocal translocation. For live birth, the risk for male carrier is 8.7-17.4%. It is important to consider this higher risk when counseling. Precise study concerning the presence of the DGCR can facilitate in the better understanding of the condition. PMID- 11491309 TI - Trisomy 6q syndrome: a case with a << de novo >> 6q23 tandem duplication. AB - In this study, we report the combined use of whole and partial chromosome 6 painting probe and YACS probes to define the unbalanced region of a de novo 6q+ marker chromosome. A male patient with peculiar features of << distal 6q trisomy syndrome >> showed a direct duplication of 6q23 region. Comparing the phenotype of this child with the phenotype of other << de novo >> partial 6q trisomy, we conclude that band 6q23 has an important role in defining 6q trisomy. PMID- 11491310 TI - De novo ring chromosome 3 in a girl with hypoplastic thumb and coloboma of iris. AB - We describe a female child with a ring chromosome 3, found after investigation for short stature. Her karyotype was 46,XX,r(3)(p26-q29). Her phenotype mainly differs from that of the nine patients previously reported with ring chromosome 3, by the presence of hypoplastic right thumb and bilateral coloboma of the iris. PMID- 11491311 TI - Extrahepatic biliary atresia with laterality sequence anomalies. AB - The patient was the first child of first cousin parents. He was born at term after an uneventful pregnancy with normal height, weight and head circumference. Jaundice appeared at 15 days of age. Ventricular septal defects and valvular pulmonary stenosis were diagnosed. An hepatic workup revealed extrahepatic biliary atresia and abdominal situs inversus. Hepatic biopsy showed cirrhosis with intrahepatic cholestasis. Genetic factors are suggested in extrahepatic biliary atresia. Analysis of segregation patterns suggested the existence of two major groups, one with various combinations of anomalies within the laterality sequence and the other with one or two anomalies mostly involving the cardiac, gastrointestinal, and urinary systems. This patient belongs to the first group. PMID- 11491312 TI - Lateral facial clefts: a case report. AB - A female new-born with bilateral lateral facial clefts, resulting in macrostomia, is reported. In addition she had a diminished palpebral fissure length. She died suddenly at the age of 6 months, presumably due to respiratory insufficiency. The unique combination of facial anomalies, present in this patient, has not been reported before. We discuss the differential diagnosis and the classification of facial clefts. PMID- 11491313 TI - Prenatal diagnosis and false negative result for a case of Down syndrome with normal karyotype. PMID- 11491315 TI - Presymptomatic testing for autosomal dominant spinocerebellar ataxia type 1 in a French family. PMID- 11491314 TI - A fetus with pseudo Fryns syndrome and t(18;22) translocation. PMID- 11491316 TI - Anophthalmia/microphtalmia associated with right lateral facial cleft/microtia. PMID- 11491317 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 11491318 TI - Notification that new names and new combinations have appeared in volume 51, part 3, of the IJSEM. PMID- 11491319 TI - Flavobacterium frigidarium sp. nov., an aerobic, psychrophilic, xylanolytic and laminarinolytic bacterium from Antarctica. AB - A psychrophilic, aerobic bacterium designated A2iT was isolated from marine sediment recovered from shallow waters surrounding Adelaide Island, Antarctica (67 degrees 34' S, 68 degrees 07' W). The organism exhibited xylanolytic and laminarinolytic activity and was halotolerant. Basic characterization showed that it was gram-negative, non-motile, yellow-pigmented (beta,beta-carotene-3,3'-diol) and positive for oxidase and catalase synthesis. Analysis of the 16S rDNA sequence suggests that the organism belongs to the Flexibacter-Cytophaga Bacteroides phylum. On the basis of its 16S rDNA sequence, the bacterium is 96.8% similar to Flavobacterium columnare ATCC 43622--its closest relation. The genomic DNA G+C content was 35 mol%. Growth on xylan occurs optimally at 15 degrees C, though growth also occurs at 0 degrees C, and the doubling times are 9.6 and 34.8 h, respectively. The maximum growth temperature on xylan is at 24 degrees C. The bacterium is a neutrophile, growing across the pH range 5.6-8.4 and having an optimum at pH 7.5. Analysis of the 16S rDNA sequence, together with phenotypic characterization, suggests that the organism is a member of the genus Flavobacterium. DNA-DNA hybridization experiments have shown that it is a novel species; it is proposed, therefore, that the organism be designated as the type strain of Flavobacterium frigidarium sp. nov. (= ATCC 700810T = NCIMB 13737T). PMID- 11491320 TI - Alkaliphilus transvaalensis gen. nov., sp. nov., an extremely alkaliphilic bacterium isolated from a deep South African gold mine. AB - A novel extreme alkaliphile was isolated from a mine water containment dam at 3.2 km below land surface in an ultra-deep gold mine near Carletonville, South Africa. The cells of this bacterium were straight to slightly curved rods, motile by flagella and formed endospores. Growth was observed over the temperature range 20-50 degrees C (optimum 40 degrees C; 45 min doubling time) and pH range 8.5 12.5 (optimum pH 10.0). The novel isolate, one of the most alkaliphilic micro organisms yet described, was a strictly anaerobic chemo-organotroph capable of utilizing proteinaceous substrates such as yeast extract, peptone, tryptone and casein. Elemental sulfur, thiosulfate or fumarate, when included as accessory electron acceptors, improved growth. The G+C content of genomic DNA was 36.4 mol %. Phylogenetic analysis based on the 16S rDNA sequence indicated that the isolate is a member of cluster XI within the low G+C gram-positive bacteria, but only distantly related to previously described members. On the basis of physiological and molecular properties, the isolate represents a novel species, for which the name Alkaliphilus transvaalensis gen. nov., sp. nov. is proposed (type strain SAGM1T = JCM 10712T = ATCC 700919T). The mechanism of generation of the highly alkaline microbial habitat and the possible source of the alkaliphile are discussed. PMID- 11491321 TI - Bordetella petrii sp. nov., isolated from an anaerobic bioreactor, and emended description of the genus Bordetella. AB - A novel Bordetella species was isolated from an anaerobic, dechlorinating bioreactor culture enriched from river sediment. The only strain, Se-1111R(T) (= DSM 12804T = CCUG 43448T), for which the name Bordetella petrii is proposed, is designated the type strain of the novel species. Strain Se-1111R(T) was isolated from the dechlorinating mixed culture due to its ability to anaerobically reduce selenate to elemental selenium. Comparative 16S rDNA sequence analysis showed a close relationship between Se-1111R(T) and members of the genus Bordetella within the beta-Proteobacteria. This close phylogenetic relatedness was also reflected in several metabolic properties of Se-1111R(T), including its incapacity to utilize carbohydrates, by the high G+C content (63.8 mol%) of its DNA and by the presence of Q-8 as the major isoprenoid quinone. DNA-DNA hybridization experiments with type strains of all species of the genus Bordetella and closely related species Achromobacter xylosoxidans subsp. denitrificans provided further evidence for the assignment of strain Se-1111R(T) as a novel species of the genus Bordetella. This genus currently consists of seven aerobic species, all of which are known to occur in close pathogenic, opportunistic or possibly commensal relationships with various host organisms. B. petrii is the first member of this genus isolated from the environment and capable of anaerobic growth. The proposal of the novel species and an emended description of the genus Bordetella is presented. PMID- 11491322 TI - Description of Microbacterium foliorum sp. nov. and Microbacterium phyllosphaerae sp. nov., isolated from the phyllosphere of grasses and the surface litter after mulching the sward, and reclassification of Aureobacterium resistens (Funke et al. 1998) as Microbacterium resistens comb. nov.. AB - The taxonomic position of a group of coryneform bacteria isolated from the phyllosphere of grasses and the surface litter after sward mulching was investigated. On the basis of restriction analyses of 16S rDNA, the isolates were divided into two genotypes. According to the 16S rDNA sequence analysis, representatives of both genotypes were related at a level of 99.2% similarity and clustered within the genus Microbacterium. Chemotaxonomic features (major menaquinones MK-12, MK-11 and MK-10; predominating iso- and anteiso-branched cellular fatty acids; G+C content 64-67 mol%; peptidoglycan-type B2beta with glycolyl residues) corresponded to this genus as well. DNA-DNA hybridization studies showed a reassociation value of less than 70% between representative strains of both subgroups, suggesting that two different species are represented. Although the extensive morphological and physiological analyses did not reveal any differentiating feature for the genotypes, differences in the presence of the cell-wall sugar mannose enabled the subgroups to be distinguished from one another. DNA-DNA hybridization with type strains of closely related Microbacterium spp. indicated that the isolates represent two individual species, which can also be differentiated from previously described species of Microbacterium on the basis of biochemical features. As a result of phenotypic and phylogenetic analyses, the species Microbacterium foliorum sp. nov., type strain P 333/02T (= DSM 12966T = LMG 19580T), and Microbacterium phyllosphaerae sp. nov., type strain P 369/06T (= DSM 13468T = LMG 19581T), are proposed. Furthermore, the reclassification of Aureobacterium resistens (Funke et al. 1998) as Microbacterium resistens (Funke et al. 1998) comb. nov. is proposed. PMID- 11491323 TI - Streptococcus australis sp. nov., a novel oral streptococcus. AB - Strains of streptococci were isolated from the mouths of children attending the United Dental Hospital, Sydney, Australia. These strains were analysed biochemically using the Rapid ID32 Strep microsystem, were subjected to DNA-DNA hybridization with other members of the oral streptococci and had their 165 rRNA analysed. On the basis of DNA-DNA hybridization, their nearest relative was Streptococcus parasanguinis, whereas, on the basis of 16S rRNA analysis, it was Streptococcus infantis. The name Streptococcus australis sp. nov. is proposed for the new species. The type strain is AI-1T (= ATCC 700641T = NCTC 13166T). PMID- 11491324 TI - Thalassomonas viridans gen. nov., sp. nov., a novel marine gamma-proteobacterium. AB - A new genus and species are proposed for two halophilic, strictly aerobic, chemo organotrophic, marine bacterial strains. These bacteria are gram-negative, motile rods isolated from oysters cultivated off the Mediterranean coast at Valencia (Spain). They produce green/blue-green diffusible pigment. The G+C content of the DNA of the proposed type strain (XOM25T) is 48.4 mol %. A 16S rRNA gene sequence analysis of the two strains has shown that the new isolates represent a branch within the gamma-Proteobacteria, close to the genus Colwellia. The type species of the new genus is Thalassomonas viridans gen. nov., sp. nov., with the type strain XOM25T (= CECT 5083T = DSM 13754T). PMID- 11491325 TI - Samsonia erythrinae gen. nov., sp. nov., isolated from bark necrotic lesions of Erythrina sp., and discrimination of plant-pathogenic Enterobacteriaceae by phenotypic features. AB - Bacterial strains isolated from diseased erythrina (Erythrina sp.) trees in Martinique (French West Indies) were studied using phenotypic tests, 16S rDNA sequence analysis and DNA-DNA hybridization. Numerical analysis of phenotypic characteristics showed that these strains formed an homogeneous phenon among plant-pathogenic Enterobacteriaceae, and gave useful and updated information for the identification of these bacteria. Results of DNA-DNA hybridization indicated that strains from erythrina belonged to a discrete genomospecies (89-100% hybridization) and had low levels of DNA relatedness (2-33% hybridization) with reference strains of phytopathogenic Erwinia, Brenneria, Pectobacterium, Pantoea and Enterobacter species. 16S rDNA sequence analysis using three different methods revealed that the position of strain CFBP 5236T isolated from erythrina was variable in the different trees, so that strains from erythrina could not be assigned to any recognized genus. It is proposed that these strains are included in a new genus, Samsonia. The name Samsonia erythrinae is proposed for the new species. The G+C content of the DNA of the type strain, CFBP 5236T (= ICMP 13937T), is 57.0 mol%. PMID- 11491326 TI - Novel nitrogen-fixing acetic acid bacteria, Gluconacetobacter johannae sp. nov. and Gluconacetobacter azotocaptans sp. nov., associated with coffee plants. AB - Diazotrophic bacteria were isolated, in two different years, from the rhizosphere and rhizoplane of coffee (Coffea arabica L.) plants cultivated in Mexico; they were designated as type DOR and type SAd isolates. They showed characteristics of the family Acetobacteraceae, having some features in common with Gluconacetobacter (formerly Acetobacter) diazotrophicus, the only known N2-fixing species of the acetic acid bacteria, but they differed from this species with regard to several characteristics. Type DOR isolates can be differentiated phenotypically from type SAd isolates. Type DOR isolates and type SAd isolates can both be differentiated from Gluconacetobacter diazotrophicus by their growth features on culture media, their use of amino acids as nitrogen sources and their carbon-source usage. These results, together with the electrophoretic mobility patterns of metabolic enzymes and amplified rDNA restriction analysis, suggested that the type DOR and type SAd isolates represent two novel N2-fixing species. Comparative analysis of the 16S rRNA sequences revealed that strains CFN-Cf55T (type DOR isolate) and CFN-Ca54T (type SAd isolate) were closer to Gluconacetobacter diazotrophicus (both strains had sequence similarities of 98.3%) than to Gluconacetobacter liquefaciens, Gluconacetobacter sacchari (similarities < 98%) or any other acetobacteria. Strain CFN-Cf55T exhibited low levels of DNA-DNA reassociation with type SAd isolates (mean 42%) and strain CFN Ca54T exhibited mean DNA-DNA reassociation of 39.5% with type DOR isolates. Strains CFN-Cf55T and CFN-Ca54T exhibited very low DNA reassociation levels, 7 21%, with other closely related acetobacterial species. On the basis of these results, two novel N2-fixing species are proposed for the family Acetobacteraceae, Gluconacetobacter johannae sp. nov. (for the type DOR isolates), with strain CFN-Cf55T (= ATCC 700987T = DSM 13595T) as the type strain, and Gluconacetobacter azotocaptans sp. nov. (for the type SAd isolates), with strain CFN-Ca54T (= ATCC 70098ST = DSM 13594T) as the type strain. PMID- 11491327 TI - DNA-DNA hybridization study of Bradyrhizobium strains. AB - DNA-DNA hybridizations were performed between Bradyrhizobium strains, isolated mainly from Faidherbia albida and Aeschynomene species, as well as Bradyrhizobium reference strains. Results indicated that the genus Bradyrhizobium consists of at least 11 genospecies, I to XI. The genospecies formed four subgeneric groups that were more closely related to each other (>40% DNA hybridization) than to other genospecies (<40% DNA hybridization): (i) genospecies I (Bradyrhizobium japonicum), III (Bradyrhizobium liaoningense), IV and V; (ii) genospecies VI and VIII; (iii) genospecies VII and IX; and (iv) genospecies II (Bradyrhizobium elkanii), X and XI. Photosynthetic Aeschynomene isolates were found to belong to at least two distinct genospecies in one subgeneric group. DNA-DNA hybridization data are compared with data from amplified fragment length polymorphism analysis and 165-23S rDNA spacer sequence analysis. PMID- 11491328 TI - Actinomyces suimastitidis sp. nov., isolated from pig mastitis. AB - An unusual Actinomyces-like bacterium originating from a pig with mastitis was subjected to a polyphasic taxonomic investigation. The morphological and biochemical characteristics of the organism were consistent with its preliminary assignment to the genus Actinomyces but it did not appear to correspond to any recognized species. PAGE analysis of whole-cell proteins confirmed the phenotypic distinctiveness of the bacterium and 16S rRNA gene sequence analysis demonstrated that it represents a hitherto unknown sub-line amongst a cluster of Actinomyces species which embraces Actinomyces canis, Actinomyces georgiae, Actinomyces hyovaginalis, Actinomyces meyeri, Actinomyces odontolyticus, Actinomyces radingae and Actinomyces turicensis. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium isolated from pig mastitis be classified as Actinomyces suimastitidis sp. nov. The type strain of Actinomyces suimastitidis is CCUG 39279T (= CIP 106779T). PMID- 11491329 TI - Thermosipho geolei sp. nov., a thermophilic bacterium isolated from a continental petroleum reservoir in Western Siberia. AB - Three strictly anaerobic, thermophilic bacteria (SL31T, SL30 and MLM39636) were isolated from a deep continental oil reservoir in Western Siberia (Russia). Following the mid-exponential phase of growth, the non-motile rod-shaped organisms were surrounded by a sheath-like structure. As DNA-DNA hybridizations showed that these strains were highly related genomically, only strain SL31T was studied in detail. The temperature range for growth of strain SL31T was between 45 and 75 degrees C, with optimum growth at 70 degrees C. Its optimum pH and NaCl concentration for growth were pH 7.5 and 20-30 g l(-1), respectively. The novel isolate reduced elemental sulfur and cystine, but not thiosulfate or sulfate, to hydrogen sulfide. The G+C content of the genomic DNA was 30.0 mol %. As determined by 16S rDNA sequence analysis, this organism belonged to the genus Thermosipho. DNA-DNA hybridization levels between strain SL31T and type strains of the previously described species of Thermosipho were less than 10%. On the basis of physiological and molecular properties, it is proposed that this organism should be placed in a new species, Thermosipho geolei sp. nov. The novel organism represents the first species of the genus Thermosipho that has been isolated from a petroleum reservoir. The type strain is SL31T ( = DSM 13256T = JCM 10986T). PMID- 11491330 TI - Thermoanaerobacter tengcongensis sp. nov., a novel anaerobic, saccharolytic, thermophilic bacterium isolated from a hot spring in Tengcong, China. AB - A new, extremely thermophilic bacterium, designated strain MB4T, was isolated from a Chinese hot spring. The new isolate was an obligately anaerobic, rod shaped, gram-negative, saccharolytic bacterium. Spore formation was not observed. Growth occurred at temperatures between 50 and 80 degrees C, with an optimum of around 75 degrees C; at pH values between 5.5 and 9.0, with an optimum of 7.0 7.5; and at salinities between 0 and 2.5% NaCl, with an optimum of around 0.2% NaCl. The organism utilized glucose, galactose, maltose, cellobiose, mannose, fructose, lactose, mannitol and starch. Acetate was the main end product from glucose fermentation. Thiosulfate and sulfur were reduced to hydrogen sulfide. Sulfate, sulfite and nitrate were not reduced. Growth was inhibited by hydrogen. The G+C content of the DNA was 33 mol%. Phylogenetic analyses based on the 16S rDNA sequence indicated that the isolate was a new member of the genus Thermoanaerobacter and formed a monophyletic unit within the Thermoanaerobacter cluster. Based on its phenotypic and phylogenetic characteristics, the isolate was proposed as a new species, Thermoanaerobacter tengcongensis. The type strain is MB4T (= Chinese Collection of Microorganisms AS 1.2430T = JCM 11007T). PMID- 11491331 TI - Comparative ribotyping of Staphylococcus intermedius isolated from members of the Canoidea gives possible evidence for host-specificity and co-evolution of bacteria and hosts. AB - A total of 41 Staphylococcus intermedius isolates were isolated from skin of healthy members of six phylogenetic groups within the Canoidea (the dog family, skunk subfamily, weasel subfamily, racoon family, red panda and bear family) of different geographical origin and compared by EcoRI ribotyping and cluster analysis. The S. intermedius isolates from the different families and subfamilies clustered together in separate groups, almost completely following the phylogenetic relationship of the animal hosts. These ribotype data indicate host specificity of different types of S. intermedius and suggest co-evolution between the animal hosts within the Canoidea and S. intermedius. PMID- 11491332 TI - Corynebacterium testudinoris sp. nov., from a tortoise, and Corynebacterium felinum sp. nov., from a Scottish wild cat. AB - Two unknown gram-positive, rod-shaped bacteria isolated from a tortoise and a Scottish wild cat were subjected to a polyphasic taxonomic analysis. Chemical analysis revealed the presence of straight-chain and monounsaturated fatty acids and short-chain mycolic acids in the two isolates consistent with the genus Corynebacterium. Comparative 16S rRNA gene sequencing confirmed that the unknown isolates were members of the genus Corynebacterium, with the two organisms displaying greater than 3% sequence divergence from each other and from established species of the genus. The unknown Corynebacterium isolates were readily distinguished from each other and from all recognized species of the genus by biochemical tests. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown organisms from a tortoise and a cat be classified in the genus Corynebacterium as Corynebacterium testudinoris sp. nov. and Corynebacterium felinum sp. nov., respectively. The respective type strains of C. testudinoris and C. felinum are CCUG 41823T and CCUG 39943T. PMID- 11491333 TI - Phylogeny of Rickettsia spp. inferred by comparing sequences of 'gene D', which encodes an intracytoplasmic protein. AB - 'Gene D' is the PS120-protein-encoding gene, first described in Rickettsia conorii and Rickettsia japonica. Sequence analysis of a 3030 bp fragment of 'gene D' in 24 representatives of the genus Rickettsia was carried out to complete phylogenetic analyses previously inferred by comparison of gene sequences encoding citrate synthase, 17 kDa antigen and rOmpA and rOmpB. The phylogenetic relationships between rickettsiae were inferred from the comparison of both the gene and the derived protein sequences, using the parsimony, neighbour-joining and maximum-likelihood methods. Five distinct groups of rickettsiae were identified. These were: the Rickettsia massiliae group, including R. massiliae, Bar 29, Rickettsia rhipicephali and Rickettsia aeschlimannii; the Rickettsia rickettsii group containing Rickettsia sibirica, 'Rickettsia mongolotimonae', Rickettsia parkeri, strain S, Rickettsia africae, the R. conorii complex, Rickettsia slovaca, Rickettsia honei, R. rickettsii, R. japonica and Rickettsia montanensis; the group currently containing only Rickettsia helvetica; the Rickettsia akari group including Rickettsia australis, R. akari and the ELB agent; Rickettsia prowazekii and Rickettsia typhi clustered in the typhus group. As significant bootstrap values were obtained for most of the nodes, sequence comparison of 'gene D' should be considered as a complementary approach in phylogenetic studies of rickettsiae. PMID- 11491334 TI - Simplified technique for identification of the aerobic spore-forming bacteria by phenotype. AB - The use of modern research approaches of genetics, biochemistry and molecular biology has led to progress in bacterial taxonomy. Systematic study of the aerobic spore-forming bacteria has resulted in the realignment of the genus Bacillus into several new genera. In the meantime, the identification process has become more difficult for the non-specialist in Bacillus taxonomy. This paper presents a key for the simplified phenotypic identification of the mesophilic, aerobic, spore-forming bacteria belonging to the genera Bacillus, Paenibacillus, Brevibacillus, Aneurinibacillus, Geobacillus and Virgibacillus. A total of 81 species were included and 115 morphological and physiological tests were analysed for their discriminative efficiency. This key is practical for rough but quick identification of aerobic spore-forming bacteria isolated from nature. Such preliminary identification will be helpful for the selection of reference strains and methods for more precise identification using the newest techniques. The reliability of the proposed identification key was tested on 100 cultures from the Ukrainian Collection of Microorganisms. The developed identification key is represented in interactive mode on a website (http://www/imv.kiev.ua/key/). PMID- 11491335 TI - Propionibacterium microaerophilum sp. nov., a microaerophilic bacterium isolated from olive mill wastewater. AB - A new gram-positive, facultative anaerobic, microaerophilic bacterium, designated strain M5T, was isolated from a decantation reservoir of olive mill wastewater. The cells were rod-shaped, non-motile, non-spore-forming and catalase-negative. Growth occurred at pH ranging from 4.5 to 9.5, with optimum growth at 7.0. The optimum temperature for growth was around 30 degrees C. Although growth occurred under anaerobic and aerobic conditions, the optimum O2 concentration for growth was determined as 5% in the gas phase of the culture. During anaerobic growth, glucose or lactate were mainly fermented to propionate, acetate and CO2. In the presence of O2 (more than 2%), glucose was oxidized completely to CO2. The G+C content of the DNA was 67.7+/-0.6 mol% and 16S rRNA gene analysis revealed that the new isolate belonged to the cluster of 'dairy' propionibacteria, Propionibacterium acidipropionici being its closest phylogenic relative (97.5% similarity). However, the level of DNA relatedness between strain M5T and P. acidipropionici was 56.2%. Consequently, both the phenotypic (range of substrates used) and genotypic characteristics of strain M5T allow it to be assigned as a new species of the genus Propionibacterium, Propionibacterium microaerophilum sp. nov. The type strain is strain M5T (= CNCM I-2360T = DSM 13435T). PMID- 11491336 TI - Vibrio shiloi sp. nov., the causative agent of bleaching of the coral Oculina patagonica. AB - The aetiological agent of bleaching of the coral Oculina patagonica was characterized as a new Vibrio species on the basis of 16S rDNA sequence, DNA-DNA hybridization data and phenotypic properties, including the cellular fatty acid profile. Based on its 16S rDNA and DNA-DNA hybridization, the new Vibrio species is closely related to Vibrio mediterranei. The name Vibrio shiloi sp. nov. is proposed for the new coral-bleaching species, the type strain being AK1T (= ATCC BAA-91T = DSM 13774T). PMID- 11491337 TI - Phylogeny of the seal mycoplasmas Mycoplasma phocae corrig., Mycoplasma phocicerebrale corrig. and Mycoplasma phocirhinis corrig. based on sequence analysis of 16S rDNA. AB - The nucleotide sequences of the 16S rRNA genes from the type strains of three seal mycoplasmas, Mycoplasma phocicerebrale, Mycoplasma phocae and Mycoplasma phocirhinis (formerly Mycoplasma phocacerebrale, Mycoplasma phocidae and Mycoplasma phocarhinis, respectively), were determined by direct DNA cycle sequencing. Polymorphisms were found in all three 16S rRNA gene sequences, showing the existence of two different rRNA operons. In M. phocae, a length difference was found between the operons, caused by an insertion or a deletion of an adenosine in one of the operons. The sequence information was used to construct phylogenetic trees. All three species were found to belong to the hominis group, but to different clusters. M. phocicerebrale and M. phocae were found to be members of the Mycoplasma hominis cluster, within which M. phocicerebrale grouped in the Mycoplasma alkalescens subcluster. M. phocirhinis was found to be a member of the Mycoplasma bovigenitalium subcluster of the Mycoplasma bovis cluster. The 16S rRNA gene sequences of all hitherto validly described species within the M. hominis and M. bovis clusters have now been determined. PMID- 11491338 TI - A physical and genetic map of the Mycoplasma flocculare ATCC 27716 chromosome reveals large genomic inversions when compared with that of Mycoplasma hyopneumoniae strain J(T). AB - Pulsed-field gel electrophoresis and DNA hybridization data were used to construct a chromosomal map of Mycoplasma flocculare ATCC 27716, a non-pathogenic inhabitant of porcine respiratory tracts. Twenty-one genetic markers were placed on the map. Comparison of the genetic map with that of the closely related Mycoplasma hyopneumoniae strain J(T), the type strain of the causative agent of enzootic pneumonia in pigs, identified three chromosomal inversions that differentiate these genomes. One of these inversions involves genes that possibly may be involved in M. hyopneumoniae pathogenicity. PMID- 11491339 TI - Facklamia miroungae sp. nov., from a juvenile southern elephant seal (Mirounga leonina). AB - An unusual gram-positive, catalase-negative, facultatively anaerobic, coccus shaped organism that originated from a juvenile elephant seal was characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing showed that the unknown coccus represents a new subline within the genus Facklamia. The unknown strain was readily distinguishable from all currently recognized species of the genus Facklamia (Facklamia hominis, Facklamia languida, Facklamia ignava, Facklamia sourekii and Facklamia tabacinasalis) by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Facklamia miroungae sp. nov. The type strain of F. miroungae is CCUG 42728T (= CIP 106764T). F. miroungae is the first member of the genus Facklamia to be isolated from an animal other than man. PMID- 11491340 TI - Proposal of the genus Sphingomonas sensu stricto and three new genera, Sphingobium, Novosphingobium and Sphingopyxis, on the basis of phylogenetic and chemotaxonomic analyses. AB - Phylogenetic analyses of 16S rRNA gene sequences by distance matrix and parsimony methods indicated that the currently known species of the genus Sphingomonas can be divided into four clusters. Some chemotaxonomic and phenotypic differences were noted among these clusters. Three new genera, Sphingobium, Novosphingobium and Sphingopyxis, are proposed in addition to the genus Sphingomonas sensu stricto. The genus Sphingobium is proposed to accommodate Sphingomonas chlorophenolica, Sphingomonas herbicidovorans and Sphingomonas yanoikuyae. The genus Novosphingobium is proposed for Sphingomonas aromaticivorans, Sphingomonas capsulata, Sphingomonas rosa, Sphingomonas stygia, Sphingomonas subarctica and Sphingomonas subterranea. Sphingomonas macrogoltabidus and Sphingomonas terrae are reclassified in the genus Sphingopyxis. The type species of Sphingobium, Novosphingobium and Sphingopyxis are Sphingobium yanoikuyae, Novosphingobium capsulatum and Sphingopyxis macrogoltabida, respectively. PMID- 11491341 TI - Nocardia cyriacigeorgici sp. nov.. AB - Chemotaxonomic and 16S rRNA gene sequence analyses of an isolate from the bronchial secretions of a patient with chronic bronchitis demonstrated clearly that it belongs to the genus Nocardia. The 16S rRNA gene sequence data, as well as the biochemical characteristics of the isolate, indicated that it belongs to a new species that differs from previously described members of the genus Nocardia. The name Nocardia cyriacigeorgici sp. nov. is proposed for this isolate, and is represented by strain IMMIB D-1627T (= DSM 44484T). PMID- 11491342 TI - Hydrogenobacter subterraneus sp. nov., an extremely thermophilic, heterotrophic bacterium unable to grow on hydrogen gas, from deep subsurface geothermal water. AB - A novel extreme thermophile was isolated from a water sample derived from a deep subsurface geothermal water pool at a depth of 1500 m in the Hacchoubaru geothermal plant in Oita Prefecture, Japan. The cells were found to be straight rods, each being motile by means of a polar flagellum. Growth was observed at temperatures between 60 and 85 degrees C (optimum 78 degrees C; 120 min doubling time) and between pH 5.5 and pH 9.0 (optimum 7.5). The isolate was a strictly aerobic heterotroph capable of utilizing a number of substrates such as yeast extract, peptone, tryptone, various carbohydrates, sugars, amino acids and organic acids. Elemental sulfur, thiosulfate, sulfide or cysteine-hydrochloride was required as an electron donor for growth. Hydrogen gas did not support growth. The G+C content of the genomic DNA was 44.7 mol%. Phylogenetic analysis based on 16S rDNA sequences and DNA-DNA hybridization analysis indicated that the isolate was closely related to members of the hydrogen-oxidizing, autotrophic and thermophilic genera Hydrogenobacter and Calderobacterium. However this isolate was differentiated from the previously described species of these genera on the basis of the physiological and molecular properties of the new isolate. The name Hydrogenobacter subterraneus sp. nov. is proposed; the type strain is HGP1T (= JCM 10560T = IFO 16485T). PMID- 11491343 TI - Nonomuraea dietzii sp. nov.. AB - The taxon 'Sebekia benihana', an actinomycete of industrial significance, has been subjected to a polyphasic taxonomic study. 16S rDNA sequence analyses of strain DSM 44320 (= NRRL 11111) revealed 100% similarity to the 16S rDNA sequence of Nonomuraea roseola DSM 43767T and high values (>98.5%) to some other species of this genus. DNA-DNA similarity values of less than 31% observed between strain DSM 44320 and N. roseola DSM 43767T, Nonomuraea recticatena DSM 43937T and Nonomuraea africana DSM 43748T indicate that this strain represents an individual genospecies. Differences between strain DSM 44320T and the 16 validly described Nonomuraea species occur in the primary structure of the 16S rDNA, the physiological test profile, the morphology of mycelium and the spore surface and the colour of substrate mycelium. The only strain of 'S. benihana', DSM 44320T (= NRRL 11111T), is proposed as the type strain of a new species of the genus Nonomuraea, Nonomuraea dietzii sp. nov.. PMID- 11491344 TI - Transfer of Halomonas canadensis and Halomonas israelensis to the genus Chromohalobacter as Chromohalobacter canadensis comb. nov. and Chromohalobacter israelensis comb. nov.. AB - 16S rRNA gene sequence comparisons and DNA-DNA hybridization data support the conclusion that two species previously described as members of the genus Halomonas, Halomonas israelensis and Halomonas canadensis, should be placed in the genus Chromohalobacter. Both H. israelensis ATCC 43985T (= Ba1T) and H. canadensis ATCC 43984T (= NRCC 41227T) have been used extensively for physiological studies for many years; nevertheless, they were not named and classified taxonomically until quite recently. Their phenotypic resemblance (at least 65% Jaccard similarity) to some members of the genus Halomonas and the degree of DNA-DNA relatedness (lower than 60%) to other described species of this genus permitted the conclusion that they were distinct species belonging to the genus Halomonas. In this study, the 16S rDNA of both species has been sequenced completely and found to share higher similarity to the available sequences of the moderately halophilic bacterium Chromohalobacter marismortui than to sequences of members of the genus Halomonas. C. marismortui is the sole species of the genus Chromohalobacter, also included in the family Halomonadaceae, and shares many phenotypic features with H. canadensis and H. israelensis. It is proposed that the two species should be renamed as Chromohalobacter canadensis comb. nov. and Chromohalobacter israelensis comb. nov. An emended description of the genus Chromohalobacter is given in order to include the features of these two species. PMID- 11491345 TI - Vibrio lentus sp. nov., isolated from Mediterranean oysters. AB - Twelve phenotypically similar marine bacteria have been studied by means of ribotyping, DNA-DNA hybridization and cultural and physiological characterization. Phylogenetic analysis has been performed of the 16S and 23S rRNA genes of two representative strains. Phylogenetically, they belong to the Vibrio/Photobacterium branch of the gamma-Proteobacteria and they share all of the properties that define the genus Vibrio. The strains represent a new Vibrio species that is phenotypically similar to Vibrio splendidus. However, resistance to the vibriostatic agent 0129 and production of acid from several carbohydrates allow differentiation between V. splendidus and the proposed new species. The DNA G+C content of the proposed type strain is 44.0 mol %. The name Vibrio lentus sp. nov. is proposed for the new species and strain 40M4T (= CECT 5110T = DSM 13757T) is the type strain. PMID- 11491346 TI - Chromohalobacter salexigens sp. nov., a moderately halophilic species that includes Halomonas elongata DSM 3043 and ATCC 33174. AB - Two strains that were originally isolated and characterized as members of the moderately halophilic species Halomonas elongata, strains DSM 3043 (= 1H11) and ATCC 33174 (= 1H15), were studied in detail. Their complete 16S rRNA sequences were determined and, when compared to sequences available from the databases, they showed a close phylogenetic relationship to Chromohalobacter marismortui. In addition, DNA-DNA hybridization experiments showed that both strains are members of the same species, but their DNA relatedness to the type strains of Halomonas elongata, ATCC 33173T, and Chromohalobacter marismortui, ATCC 17056T, is very low. Phenotypically, the two strains showed very similar features, related to those of Chromohalobacter, but clear differences were found between these two strains and Chromohalobacter marismortui. On the basis of these data, it is proposed that Halomonas elongata DSM 3043 and ATCC 33174 should be included in a new species of the genus Chromohalobacter, Chromohalobacter salexigens sp. nov. The type strain is DSM 3043T (= ATCC BAA-138T = CECT 5384T = CCM4921T = CIP106854T = NCIMB 13768T). PMID- 11491347 TI - Limnobacter thiooxidans gen. nov., sp. nov., a novel thiosulfate-oxidizing bacterium isolated from freshwater lake sediment. AB - Two novel thiosulfate-oxidizing strains were isolated from sediment of the littoral zone of a freshwater lake (Lake Chiemsee, Bavaria, Germany). The new isolates, designated CS-K1 and CS-K2T, were gram-negative, slightly curved rods with pointed ends that were motile by means of single polar flagella. Both strains were obligately aerobic and grew on a variety of organic substrates, but not autotrophically. The utilization of thiosulfate led to an increase in the growth yield, indicating that these strains were able to grow chemolithoheterotrophically by oxidation of thiosulfate to sulfate. The optimum thiosulfate concentrations for growth were determined to be 10 mM for strain CS K1 and 20 mM for strain CS-K2T. Phylogenetically, both strains were affiliated to the beta-Proteobacteria. Their characterization by a polyphasic approach resulted in the placement of both strains into a single species that is related only distantly to any known type species. Thus, the creation of a novel taxon is proposed, with the name Limnobacter thiooxidans gen. nov., sp. nov., to include the novel strains. In addition, the phylogenetic position of the chemolithoheterotrophic strain 'Thiobacillus' Q was determined. PMID- 11491348 TI - Description of Tropheryma whipplei gen. nov., sp. nov., the Whipple's disease bacillus. AB - A detailed characterization was performed of the Whipple's disease bacillus, strain Twist-MarseilleT, isolated from the cardiac valve of a patient with Whipple's disease bacillus endocarditis. This strain was isolated and maintained on human embryonic lung fibroblast monolayers, but could not be cultivated in the absence of living eukaryotic cells. Two morphological forms were observed, with differing staining properties; an intracellular form with intact and degenerating bacteria within vacuoles of infected cells and an extracellular form with masses of bacteria embedded in an extracellular matrix. Determination of the DNA G+C content confirmed that it belongs to the high-G+C gram-positive bacteria. Strain Twist-MarseilleT (= CNCM I-2202T) is proposed as the type strain of a new species within a new genus, Tropheryma whipplei gen. nov., sp. nov., that was provisionally created solely on the basis of 16S rRNA gene sequence data. PMID- 11491349 TI - Burkholderia ambifaria sp. nov., a novel member of the Burkholderia cepacia complex including biocontrol and cystic fibrosis-related isolates. AB - A polyphasic taxonomic study, including amplified fragment length polymorphism (AFLP) fingerprinting, DNA-DNA hybridizations, DNA base-ratio determinations, phylogenetic analysis, whole-cell fatty acid analyses and an extensive biochemical characterization, was performed on 19 Burkholderia cepacia-like isolates from the environment and cystic fibrosis (CF) patients. Several of the environmental isolates have attracted considerable interest due to their biocontrol properties. The polyphasic taxonomic data showed that the strains represent a new member of the B. cepacia complex, for which the name Burkholderia ambifaria sp. nov. is proposed. The type strain is strain LMG 19182T. B. ambifaria can be differentiated from the other members of the B. cepacia complex by means of AFLP fingerprinting, whole-cell fatty acid analysis, biochemical tests (including ornithine and lysine decarboxylase activity, acidification of sucrose and beta-haemolysis) and a newly developed recA gene-based PCR assay. 16S rDNA-based RFLP analysis and PCR tests allowed differentiation of B. ambifaria from Burkholderia multivorans, Burkholderia vietnamiensis and B. cepacia genomovar VI, but not from B. cepacia genomovars I and III and Burkholderia stabilis. The finding that this new taxon includes both strains isolated from CF patients and potentially useful biocontrol strains supports the general consensus that the large-scale use of biocontrol strains belonging to the B. cepacia complex would be ill-advised until more is known about their potential pathogenic mechanisms. PMID- 11491350 TI - Sphingomonas aquatilis sp. nov., Sphingomonas koreensis sp. nov., and Sphingomonas taejonensis sp. nov., yellow-pigmented bacteria isolated from natural mineral water. AB - The taxonomic status of four strains from several mineral water sources in Taejon, Korea, was examined. The DNA base compositions of these strains ranged from 62 to 66 mol %. All of the strains contained sphingolipids and ubiquinone 10 as the main respiratory quinone. The cellular fatty acids of these strains included octadecenoic acid, hexadecanoic acid and 2-hydroxymyristic acid, without 3-hydroxy fatty acids. On the basis of morphological, physiological and chemotaxonomic characteristics, together with 16S rDNA sequence comparison and DNA-DNA reassociation data, three new species of the genus Sphingomonas are proposed for the strains isolated from natural mineral water: Sphingomonas aquatilis sp. nov. for strain JSS7T (= KCTC 2881T = KCCM 41067T), Sphingomonas koreensis sp. nov. for strains JSS26T (= KCTC 2882T = KCCM 41069T) and JSS28 (= KCTC 2883 = KCCM 41066) and Sphingomonas taejonensis sp. nov. for strain JSS54T (= KCTC 2884T = KCCM 41068T). PMID- 11491351 TI - Pseudoalteromonas ulvae sp. nov., a bacterium with antifouling activities isolated from the surface of a marine alga. AB - A dark-purple marine bacterium that inhibits the germination of marine algal spores and the settlement of invertebrate larvae has been characterized and assessed for taxonomic assignment. Two strains, designated UL12T and UL13, were isolated from the surface of the common marine alga Ulva lactuca. Based on 16S rDNA sequencing, UL12T and UL13 were found to show the highest similarity (97%) to members of the genus Pseudoalteromonas. DNA-DNA hybridization studies demonstrated less than 28% genomic DNA relatedness between these isolates and closely related Pseudoalteromonas species and greater than 65% homology between UL12T and UL13. The two isolates were found to display identical characteristics and are strict aerobes, motile by means of single polar flagella, exhibit non fermentative metabolism and require sodium ions for growth. The isolates hydrolyse gelatin and can utilize citrate, maltose, mannose and glucose but not trehalose, sucrose, fructose, lactose or glycerol as sole carbon sources. The molecular evidence together with the phenotypic characteristics show that this bacterium constitutes a new species within the genus Pseudoalteromonas. The name Pseudoalteromonas ulvae sp. nov. is proposed for this bacterium and the type strain is UL12T (= UNSW 095600T = NCIMB 13762T). PMID- 11491352 TI - Pseudonocardia kongjuensis sp. nov., isolated from a gold mine cave. AB - The taxonomic position of an isolate that was recovered from a gold mine cave near Kongju, Republic of Korea, was determined by 16S rDNA sequence studies and chemotaxonomic characterization. Comparative studies of 16S rDNA sequences indicated that this organism was phylogenetically related to members of the genus Pseudonocardia, branching outside a cluster encompassing Pseudonocardia autotrophica and Pseudonocardia compacta. The affiliation to the genus was also supported by the cell chemistry, which was represented by a type IV cell wall, MK 8(H4) as the major menaquinone, a phospholipid type PIII pattern (phosphatidylcholine as a diagnostic phospholipid) and a DNA G+C content of 71 mol%. The fatty acid profile contained saturated, unsaturated and 10-methyl branched fatty acids, but tuberculostearic acid and hydroxy fatty acids were not present. The isolate differed from its phylogenetic neighbours in the presence of phosphatidylethanolamine, dodecanoate, 16-methylheptadecenoate and 16 methylheptadecanoate and the absence of phosphatidylinositol mannoside and phosphatidylmethylethanolamine. The unique combination of physiological properties, the cellular fatty acid profile and DNA-DNA hybridization data indicates that this organism is readily differentiated from the type strains of all of the validly published species of the genus Pseudonocardia. The name Pseudonocardia kongjuensis sp. nov. is proposed for the type strain, LM 157T (= IMSNU 50583T = KCTC 9990T = DSM 44525T). PMID- 11491353 TI - Planomicrobium koreense gen. nov., sp. nov., a bacterium isolated from the Korean traditional fermented seafood jeotgal, and transfer of Planococcus okeanokoites (Nakagawa et al. 1996) and Planococcus mcmeekinii (Junge et al. 1998) to the genus Planomicrobium. AB - A bacterial strain, JG07T, isolated from the Korean traditional fermented seafood jeotgal, was subjected to a polyphasic taxonomic study. Cells of strain JG07T are cocci or short rods in the early growth phase but change to rods as the cultures age. The peptidoglycan type is A4alpha, based on L-Lys-D-Glu. The menaquinone profile is characterized by the predominance of MK-8 followed by MK-7 and MK-6. The cellular fatty acid profile contains major amounts of saturated, unsaturated and branched fatty acids. The cellular phospholipids are phosphatidylethanolamine, phosphatidylglycerol and bisphosphatidylglycerol. The G+C content of the DNA is 47 mol%. Phylogenetic analysis showed that strain JG07T forms a cluster with Planococcus okeanokoites and Planococcus mcmeekinii, and the relationship between this cluster and two other Planococcus species described previously is supported by bootstrap analysis at a confidence level of 100%. The 16S-23S internally transcribed spacer (ITS) sequence similarity and DNA-DNA relatedness values between strain JG07T and the type strains of other Planococcus species are in the range 74.6-83.2% and 10.4-20.5%, respectively. On the basis of the phenotypic and phylogenetic data and the genomic distinctiveness, strain JG07T is considered to represent a new genus and a new species, for which the name Planomicrobium koreense gen. nov., sp. nov. is proposed. It is also proposed that Planococcus okeanokoites and Planococcus mcmeekinii be transferred to the new genus Planomicrobium as Planomicrobium okeanokoites and Planomicrobium mcmeekinii, respectively. PMID- 11491354 TI - Proposal of the genera Anaerococcus gen. nov., Peptoniphilus gen. nov. and Gallicola gen. nov. for members of the genus Peptostreptococcus. AB - Members of genus Peptostreptococcus have previously been found to be distantly related to the type species, Peptostreptococcus anaerobius, on the basis of 16S rDNA sequence similarities. They were divided into three major phylogenetic groups, and their peptidoglycan structure and biochemical traits differed between groups. The reclassification of the species of these three groups into three new genera, Peptoniphilus gen. nov., Anaerococcus gen. nov. and Gallicola gen. nov., is proposed. The genus Peptoniphilus gen. nov. includes the following butyrate producing, non-saccharolytic species that use peptone and amino acids as major energy sources: Peptoniphilus asaccharolyticus comb. nov. (type species), Peptoniphilus lacrimaris comb. nov., Peptoniphilus harei comb. nov., Peptoniphilus indolicus comb. nov. and Peptoniphilus ivorii comb. nov. The genus Anaerococcus gen. nov. contains the saccharolytic, butyrate-producing species Anaerococcus prevotii comb. nov. (type species), Anaerococcus tetradius comb. nov., Anaerococcus lactolyticus comb. nov., Anaerococcus hydrogenalis comb. nov., Anaerococcus vaginalis comb. nov. and Anaerococcus octavius sp. nov. The genus Gallicola gen. nov. contains a single species, Gallicola barnesae comb. nov. PMID- 11491355 TI - Agromyces luteolus sp. nov., Agromyces rhizospherae sp. nov. and Agromyces bracchium sp. nov., from the mangrove rhizosphere. AB - The taxonomic positions of four strains isolated from the mangrove rhizosphere were studied by a polyphasic approach using phenotypic, chemotaxonomic and genetic methods. The four isolates contain 2,4-diaminobutyric acid in their peptidoglycan, and rhamnose as the major cell wall sugar. The predominant menaquinones are MK-12 and MK-11. The predominant cellular fatty acids are iso C16:0, anteiso-C15:0 and/or anteiso-C17:0. The G+C content of the DNA ranges from 70.0 to 73.3 mol%. The four strains formed a coherent cluster with Agromyces species in a phylogenetic inference based on 16S rDNA sequences. Interestingly, the four isolates grew well in the presence of 5% NaCl. The differences in some phenotypic and chemotaxonomic characteristics, 16S rDNA sequence similarity data and DNA-DNA relatedness data indicate that the four isolates represent three new species in the genus Agromyces, for which are proposed the names Agromyces luteolus for strain 8T (IFO 16235T = VKM Ac-2085T), Agromyces bracchium for strain 65T (IFO 16238T = VKM Ac-2088T) and Agromyces rhizospherae for strains 14T (IFO 16236T = VKM Ac-2086T) and 58(5) (IFO 16237 = VKM Ac-2087). PMID- 11491356 TI - Thermoanaerobacter yonseiensis sp. nov., a novel extremely thermophilic, xylose utilizing bacterium that grows at up to 85 degrees C. AB - A novel strictly anaerobic, extremely thermophilic, spore-forming and xylose utilizing bacterium, designated strain KB-1TP (type and patent strain), was isolated from a geothermal hot stream at Sileri on Java island, Indonesia. The cells were rod-shaped, motile and had terminal spores. The newly isolated strain stained gram-positive and the cells occurred singly or in pairs during the exponential growth phase. The temperature optimum for growth was 75 degrees C and growth occurred in the range 50-85 degrees C. The pH range for growth was 4.5 9.0, with an optimum at pH 6.5. Strain KB-1TP grew chemo-organotrophically by fermenting a wide range of substrates such as glucose, fructose, D-xylose, lactose, maltose, sucrose, mannose, galactose, cellobiose, pullulan and soluble starch. Arabinose, xylan, cellulose, olive oil and Tween 80 were not fermented. The predominant fermentation end products after growth on glucose were lactate, acetate, ethanol, CO2 and small amounts of isovaleric acid, butyric acid, propionic acid, 1-pentanol and 2-propanol. Thiosulfate was reduced to H2S. Strain KB-1TP was sensitive to tetracycline, chloramphenicol, penicillin G, neomycin, kanamycin, vancomycin and rifampicin at concentrations of 100 microg ml(-1). No effect was observed with chloramphenicol and neomycin at concentrations of 10 microg ml(-1). This indicates that strain KB-1TP belongs to the bacterial domain. The G+C content of the DNA was 37 mol%. The comparison of the 165 rDNA sequence to that of closely related strains revealed that strain KB-1TP belonged to clostridial cluster V, showing highest sequence identities (92.7%) to members of the genus Thermoanaerobacter. Taking into account the physiological and molecular properties of the new isolate, it is proposed that strain KB-1TP should be classified as a new species of the genus Thermoanaerobacter, designated Thermoanaerobacter yonseiensis. The type strain, KB-1TP, has been deposited in the Korean Federation of Culture Collections (KFCC 11116P) as a patent strain and in the Deutsche Sammlung von Mikroorganismen und Zellkulturen as a type strain (= DSM 13777T). PMID- 11491357 TI - Pseudomonas kilonensis sp. nov., a bacterium isolated from agricultural soil. AB - A total of 131 bacterial isolates related to Pseudomonas corrugata were obtained from an agricultural soil from northern Germany. Based on 16S rRNA gene sequences, PCR-based genome fingerprinting and multilocus enzyme electrophoresis, they formed two groups, A (119 strains) and B (12 strains). As members of each group were highly similar, a single strain of each group was subsequently characterized by a polyphasic taxonomic approach. The selected member of group A was identified as a strain of Pseudomonas brassicacearum, whereas the selected member of group B was distinct from other species of the genus Pseudomonas. Although DNA-DNA hybridization suggested a close affiliation of the group B strain with P. brassicacearum and Pseudomonas thivervalensis and ribotyping suggested a close affiliation with P. brassicacearum, RAPD data, 16S rRNA gene sequencing and phenotypic characterization indicated the presence of a distinct taxonomic entity. This strain differed from the type strains of P. thivervalensis and P. brassicacearum in 10 and 12 metabolic properties, respectively, whereas the two organisms differ from one another by only two properties. Strains of group B are therefore considered to be members of a new species, for which the name Pseudomonas kilonensis sp. nov. is proposed. The type strain is strain 520 20T (= DSM 13647T = CFBP 5372T). PMID- 11491358 TI - Bartonella schoenbuchii sp. nov., isolated from the blood of wild roe deer. AB - The genus Bartonella comprises two human-specific pathogens and a growing number of zoonotic or animal-specific species. Domesticated as well as wild mammals can serve as reservoir hosts for the zoonotic agents and transmission to humans may occur by blood sucking arthropods or by direct blood to blood contact. Humans may come into intimate contact with free-ranging mammals during hunting, especially during evisceration with bare hands, when accidental blood to blood contact frequently occurs. The objective of this work was to determine the presence and the polymorphism of Bartonella strains in wild roe deer (Capreolus capreolus) as the most widely spread game in Western Europe. We report the isolation of four Bartonella strains from the blood of five roe deer. These strains carry polar flagella similar to Bartonella bacilliformis and Bartonella clarridgeiae. Based on their phenotypic and genotypic characteristics, three of the four roe deer isolates were different and they were all distinct from previously described Bartonella species. They can be distinguished from each other and from other Bartonella species by their protein profile, ERIC-PCR pattern, 16S rRNA and citrate synthase (gitA) gene sequences, as well as by whole DNA-DNA hybridization. In spite of their considerable heterogeneity, all four strains fulfil the criteria for belonging to a single new species. The name Bartonella schoenbuchii is proposed for this new species. The type strain R1T of Bartonella schoenbuchii has been deposited in the National Collection of Type Cultures as NCTC 13165T and the Deutsche Sammlung von Mikroorganismen und Zellkulturen as DSM 13525T. PMID- 11491359 TI - Enterococcus haemoperoxidus sp. nov. and Enterococcus moraviensis sp. nov., isolated from water. AB - A polyphasic taxonomic approach was used to study atypical enterococci isolated from surface waters. All strains were characterized by physiological and biochemical tests as well as by genotyping. The results of biochemical tests and tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all studied strains uniformly into two groups. Because these groups were clearly separated from all enterococcal species described to date, 16S rDNA sequence analysis, DNA base composition analysis and DNA-DNA hybridization of representative strains were done to elucidate the taxonomic position of the analysed groups. On the basis of the results obtained, the names Enterococcus haemoperoxidus (type strain CCM 4851T = LMG 19487T) and Enterococcus moraviensis (type strain CCM 4856T = LMG 19486T) are proposed for the two hitherto undescribed species. The type strains and reference cultures have been deposited in the Czech Collection of Microorganisms (CCM), Masaryk University, Brno, Czech Republic, and in the BCCM/LMG Culture Collection, Ghent University, Belgium. PMID- 11491360 TI - Crossiella gen. nov., a new genus related to Streptoalloteichus. AB - Phylogenetic analysis of the genera within the suborder Pseudonocardineae based on almost complete sequences of 16S rDNA showed that Saccharothrix cryophilis NRRL B-16238T was misplaced within the genus Saccharothrix. Saccharothrix cryophilis NRRL B-16238T appeared to be phylogenetically closest to Streptoalloteichus, but is morphologically distinct from this genus because sporangia with motile spores are not observed. The aerial mycelium fragments into rod-shaped elements and sclerotium-like bodies are observed occasionally in the substrate mycelium. The cell wall contains meso-diaminopimelic acid, whole-cell hydrolysates contain galactose, rhamnose and ribose, the phospholipid pattern is type PIV and the principal menaquinone is MK-9(H4). A new genus to accommodate Saccharothrix cryophilis is proposed, Crossiella gen. nov., in recognition of the contributions of Thomas Cross, a distinguished actinomycete biologist at the University of Bradford, UK. The type species is Crossiella cryophila gen. nov., comb. nov. PMID- 11491361 TI - Phylogeny of six naviculoid diatoms based on 18S rDNA sequences. AB - 18S rDNA sequences of six Naviculaceae species [Amphora montana, Gomphonema parvulum, Eolimna minima (syn. Navicula minima), Eolimna subminuscula (syn. Navicula subminuscula), Navicula veneta and Phaeodactylum tricornutum] were determined in order to assess the monophyly of this important group of diatoms, to date not included in 18S rDNA databases, and also that of the recently described genus Eolimna. Phylogenetic trees were constructed using other known diatom 18S rDNA sequences, and best tree topologies obtained were tested against alternative trees for their reliability. The analyses do not reject the monophyly of Naviculaceae and strongly support the separation of the genus Eolimna from Navicula sensu lato. The two species of Eolimna, however, do not appear to be each other's closest relatives among the species investigated: rather, E. subminuscula shows affinities to A. montana, and E. minima to P. tricornutum. A. montana, a species which it has been proposed should be transferred into a separate taxon from the other five species, was found to have grouped well within them in all analyses. PMID- 11491362 TI - Evolution of nuclear dualism in ciliates: a reanalysis in light of recent molecular data. AB - Ciliates are defined by the presence of dimorphic nuclei. Following conjugation, both the functional macronucleus (MAC) and the transcriptionally inactive germline micronucleus (MIC) develop from a zygotic nucleus. As the MAC develops, germline chromosomes are processed by excision of internal sequences, fragmentation and amplification of the remaining chromosomes. The extent of processing varies among lineages and, in all but one class of ciliates, the resulting MACs divide by an unusual process termed 'amitosis'. Research on these chromosomal rearrangements, largely from studies of only a handful of taxa from two of the nine classes of ciliates, has failed to find evidence of homologous processing among ciliate lineages. This observation, coupled with the structural diversity of MAC genomes among ciliates, led to the hypothesis of multiple origins of at least two MAC properties: (1) the ability to divide and (2) the mechanisms underlying chromosomal processing. Applying this logic to a more inclusive analysis of ciliate lineages, where an even greater diversity of MAC structure is observed, increases the potential number of origins of these MAC characteristics. Here, it is proposed that a single origin of a relatively plastic mechanism underlying MAC development better explains the observed diversity in MAC structure and processing among ciliates. Such a mechanism is suggested by the demonstration of epigenetic effects during MAC development in Paramecium and Tetrahymena. PMID- 11491363 TI - Partial sequence analysis of the actin gene and its potential for studying the phylogeny of Candida species and their teleomorphs. AB - The actin gene has been studied as a potential phylogenetic marker for selected members of the anamorphic genus Candida and seven related teleomorphic genera (Debaryomyces, Issatchenkia, Kluyveromyces, Saccharomyces and Pichia from the Saccharomycetaceae; Clavispora and Metschnikowia from the Metschnikowiaceae). The nucleotide sequences of 36 fungal taxa were analysed with respect to their molecular evolution and phylogenetic relationships. A total of 460 bp (47%) of the coding 979 bp were variable and 396 bp (40%) of these were found to be phylogenetically informative. Further analysis of the sequences showed that the genic G+C contents were higher than the nuclear G+C contents for most of the taxa. A strong positive correlation was found between G+C content over all codon positions and third positions. First and second codon positions were considered to be independent of the genic G+C content. The expected transition/transversion bias was detected only for third positions. Pairwise comparisons of transitional and transversional changes (substitutions) with total percentage sequence divergences revealed that the third position transitions showed no saturation for ingroup comparisons. A specific weighting scheme was set up, combining codon position weights with change-frequency weights to enable the inclusion of distant outgroup taxa. Parsimony analyses of the investigated taxa showed four groups, three of which corresponded to major clusters that had been established previously in Candida by rDNA analysis. Interrelationships among the species groups in this heterogeneous anamorphic genus were determined. The polyphyletic origin of the selected Candida species and their close associations with several ascomycete genera were verified and known anamorph/teleomorph pairs confirmed. The actin gene was established as a valuable phylogenetic marker with the particular advantage of an unambiguous alignment. PMID- 11491364 TI - Analysis of the constitution of the beer yeast genome by PCR, sequencing and subtelomeric sequence hybridization. AB - The lager brewing yeasts, Saccharomyces pastorianus (synonym Saccharomyces carlsbergensis), are allopolyploid, containing parts of two divergent genomes. Saccharomyces cerevisiae contributed to the formation of these hybrids, although the identity of the other species is still unclear. The presence of alleles specific to S. cerevisiae and S. pastorianus was tested for by PCR/RFLP in brewing yeasts of various origins and in members of the Saccharomyces sensu stricto complex. S. cerevisiae-type alleles of two genes, HIS4 and YCL008c, were identified in another brewing yeast, S. pastorianus CBS 1503 (Saccharomyces monacensis), thought to be the source of the other contributor to the lager hybrid. This is consistent with the hybridization of S. cerevisiae subtelomeric sequences X and Y' to the electrophoretic karyotype of this strain. S. pastorianus CBS 1503 (S. monacensis) is therefore probably not an ancestor of S. pastorianus, but a related hybrid. Saccharomyces bayanus, also thought to be one of the contributors to the lager yeast hybrid, is a heterogeneous taxon containing at least two subgroups, one close to the type strain, CBS 380T, the other close to CBS 395 (Saccharomyces uvarum). The partial sequences of several genes (HIS4, MET10, URA3) were shown to be identical or very similar (over 99%) in S. pastorianus CBS 1513 (S. carlsbergensis), S. bayanus CBS 380T and its close derivatives, showing that S. pastorianus and S. bayanus have a common ancestor. A distinction between two subgroups within S. bayanus was made on the basis of sequence analysis: the subgroup represented by S. bayanus CBS 395 (S. uvarum) has 6-8% sequence divergence within the genes HIS4, MET10 and MET2 from S. bayanus CBS 380T, indicating that the two S. bayanus subgroups diverged recently. The detection of specific alleles by PCR/RFLP and hybridization with S. cerevisiae subtelomeric sequences X and Y' to electrophoretic karyotypes of brewing yeasts and related species confirmed our findings and revealed substantial heterogeneity in the genome constitution of Czech brewing yeasts used in production. PMID- 11491365 TI - Combined intrauterine and ovarian pregnancy after in vitro fertilization and embryo transfer: a case report. AB - We describe the natural course and the management of a very rare combined intrauterine and ovarian pregnancy after IVF/ET. The rarity of heterotopic and ovarian pregnancies, with the etiologic, diagnostic and therapeutic aspects of this rare case are reported. PMID- 11491367 TI - Bilateral, tubal pregnancy treated with conservative endoscopic surgery. AB - Bilateral tubal pregnancies are very rare and those diagnosed during surgery, and at the same gestational age, are even more rare. A 28-year-old womand who had been treated with conservative laparoscopy for another tubal pregnancy a few years before and lateral by laparotomy for corpus luteal hemorrage had a bilateral tubal pregnancy treated by conservative endoscopic surgery. PMID- 11491366 TI - Presence of LH in gonadotropins associated with higher IVF pregnancy rates when basal serum LH is increased. AB - PURPOSE: To determine if pregnancy rates following in vitro fertilization-embryo transfer (IVF-ET) correlate with the presence or not of luteinizing hormone (LH) in the gonadotropins used for stimulation. Furthermore to see if the early follicular phase serum LH level affects pregnancy outcome according to the type of gonadotropins used. METHODS: The type of gonadotropins were prescribed randomly according to finances and convenience. Serum LH was obtained on day 2 or 3 of the menstrual cycle. RESULTS: When LH was > the median, significantly higher pregnancy rates were obtained in those treated with the follicle stimulating hormone (FSH)/human menopausal gonadotropin combination. When LH was < or = the median, significantly more oocytes were retrieved with FSH exclusively. No confounding variables were found to explain the data. CONCLUSIONS: Considering concerns of published studies that LH may have a toxic effect on pregnancy outcome, and if LH is suppressed too low, gonadotropins with exclusive FSH may not stimulate sufficient oocytes, the results were opposite to expectations. PMID- 11491368 TI - Adenoid cystic carcinoma of Bartholin's gland: a case report. AB - Adenoid cystic carcinoma is an uncommon histological type of the already rare carcinoma of Bartholin's gland with 51 cases described in the literature. We present a case of a 66-year-old woman who was admitted with severe pelvic pain. In an examination under anesthesia a 10x5 cm apparently fixed mass at the left vaginal wall originating from the area of Bartholin's gland was found. and biopsy indicated carcinoma. The patient underwent wide local excision. Although clinically inoperable, the tumor did not infiltrate the bony pelvis and no evidence of metastasis was found. Pathology examination revealed adenoid cystic carcinoma of Bartholin's gland. PCR did not detect human papillomavirus DNA in the specimen. The patient has been treated with adjuvant radiotherapy, and is alive with no evidence of disease after ten months. PMID- 11491369 TI - Voluminous ovarian cystoma in term pregnancy: case report. AB - Treatment of a large voluminous serous cystoadenofibroma during a cesarean section is described. The relative frequency of such pathology in pregnancy and the importance of early diagnosis are discussed. PMID- 11491370 TI - The successful use of hatched blastocysts in assisted reproductive technology. AB - This retrospective study was undertaken to determine the value of blastocyst culture and transfer as a tool in assisted reproductive technology. Six hundred and fifty-five cycles in patients undergoing IVF treatment for infertility were involved. All patients were aged < 40 years. Day-2 embryos were transferred to 427 (group 1) and day-6 embryos (blastocysts) were transferred to 228 patients (group 2). Pronucleate oocytes obtained from IVF were cultured in vitro for 2 or 6 days. One to five embryos were transferred. A total of 10,146 oocytes were retrieved, 6,105 oocytes were fertilized, 2,222 embryos were transferred and 197 clinical pregnancies were achieved in all groups. Blastocystes were transferred to almost 90% of group 2 patients. The pregnancy rate per cycle and implantation rate per transferred embryo was 42.1% and 19.4%, respectively, in the blastocyst group compared to 23.6% and 8.6%, respectively, when embryos were transferred on day 2. Even though in the blastocyst group there was an increased number of oocytes fertilized at the same time there was a significant reduction in the number of embryos being replaced (3.2 vs 3.8). This study demonstrate that transfer of blastocysts increases the success of IVF when compared with day-2 transfers and reduces the number of embryos to be transferred. PMID- 11491371 TI - Unscarred uterine rupture after induction of labor with misoprostol: a case report. AB - The rupture of an unscarred uterus is very rare and presents an emergency situation that threatens the life of the fetus and mother. The agents used for induction of labor, like oxytocin and/or prostaglandins, can be responsible for this catastrophic event. We report a case of intrapartum rupture of an intact uterus after using intravaginal misoprostol for cervical ripening and labor induction in a term pregnancy and we discuss the other cases reported in the literature. PMID- 11491372 TI - Abdominal wall endometriosis--ultrasound research: a diagnostic problem. AB - Abdominal wall endometriosis (AWE) is a rare event. Only a few reports in the literature mention sonographic features of this clinical entity. We describe a case of a young woman with subcutaneous endometriosis under the surgical scar of a previous cesarean section. Physical examination, ultrasound findings, histopathological features and differential diagnostic problems are discussed. Ultrasound examination, in combination with clinical history, is a useful method in the diagnosis of abdominal wall endometriosis and the avoidance of diagnostic pitfalls. PMID- 11491373 TI - Use of continuous fluid drainage for severe polyhydramnios due to twin to twin transfusion syndrome. AB - Acute polyhydramnios due to twin to twin transfusion is a rare complication of twin pregnancies which, despite treatment, has a high perinatal mortality. Repeated decompression amniocentesis has been used but is associated with certain risks. We report the use of continuous, gradual fluid drainage as an alternative method of uterine decompression. PMID- 11491374 TI - Comparison of the resorbable barrier interceed (TC7) and preoperative use of medroxyprogesterone acetate in postoperative adhesion prevention. AB - PURPOSE: The purpose of this study was to compare the effectiveness of barrier agent Interceed (TC7) (Johnson & Johnson Medical, Inc) and preoperative use of commonly used progesterone, medroxyprogesterone acetate. METHODS: Forty-five Spraque-Dawley white rats of reproductive age (225-250 g) were employed as a model for postsurgical adhesion formation. Group 1 consisted of control rats, Group 2 consisted only of rats with Interceed applied to the denuded areas, Group 3 consisted of rats where preoperative MPA was used. Fifteen rats were exposed to daily IM injections of 15 mg medroxyprogesterone acetate (MPA) two weeks before the surgery. RESULTS: The total adhesion score of the MPA group was significantly less than the control and Interceed group respectively (chi2=10.15) (p<0.001), (chi2=4.67) (p<0.03). There was no significant difference between the Interceed and the control group (p>0.05). CONCLUSION: Preoperative long-term MPA treatment significantly decreases primary adhesion formation. It seems that there are some other mechanisms responsible for this effect rather than anti-inflammation and/or immunosuppression. It may be that the hypoestrogenic milieu depends on the use of progesterone. PMID- 11491375 TI - Pregnancy following transfer of cryopreserved-thawed embryos that had been a result of fertilization of all in vitro matured metaphase or germinal stage oocytes. case report. AB - PURPOSE: To see if pregnancies can be achieved after performing intracytoplasmic sperm injection (ICSI) of in vitro matured metaphase I or germinal vesicle stage oocytes and then cryopreserving them at the 2 pronuclear stage. METHODS: Transfer of frozen/thawed hatched day 3 embryos to two women. All embryos were a result of in vitro maturation of immature oocytes followed by ICSI. RESULTS: Both women conceived. One has delivered and one has successfully completed the first trimester. CONCLUSION: These two cases represent only the second and third reported cases of pregnancies following frozen embryo transfer where the embryos resulted from fertilizing immature oocytes by ICSI. PMID- 11491376 TI - Salpingectomy improves outcome in the presence of a unilateral hydrosalpinx in a donor oocyte recipient: a case report. AB - PURPOSE: To determine if unilateral salpingectomy for hydrosalpinx can improve fecundity in a woman with many cycles of failure to conceive despite the fertilization of donor oocytes and subsequent embryo transfer. METHODS: Salpingectomy performed after failure to conceive despite IVF-ET with the patient's own oocytes (n=5) or transfer of donor embryos (n=2) or embryo transfer cycles as a donor oocyte recipient (n=5). RESULTS: The patient conceived three of four times following salpingectomy. Transfer of four frozen-thawed donor embryos and two frozen-thawed embryos of her own led to a successful delivery. In one of two cycles as a donor oocyte recipient she had a successful delivery and subsequently, the transfer of cryopreserved/thawed embryos from a previous donor oocyte cycle led to a chemical pregnancy. CONCLUSION: Unilateral hydrosalpinx can be a cause of recalcitrant failure to conceive despite assisted reproductive technology. Salpingectomy can restore fecundity. PMID- 11491377 TI - Pregnancy rates (prs) according to embryo cell number at time of embryo transfer (ET). AB - PURPOSE: To evaluate pregnancy and implantation rates following fresh and frozen embryo transfer (ET) according to blastomere number. METHODS: A retrospective study from 1/1/97 to 9/30/98 including all cycles with ETs irrespective of age. RESULTS: 65% of fresh transfers had at least one 8-cell embryo vs only 39.6% for frozen ET. The clinical pregnancy and implantation rates were higher when one 8 cell embryo was transferred (64% and 24%) vs a 5-7 cell embryo (41% and 14.5%) for fresh transfers. There was less of a difference with frozen ETs (46% and 19% for 8-cell vs 38% and 17% for 5-7 cell). CONCLUSIONS: Since mostly only 8-cell embryos at day 3 reach the blastocyst stage, these data raise questions as to whether the quest to attain the highest pregnancy rate per transfer through blastocyst transfer, may be at the expense of overall pregnancy rate (fresh and frozen) from a given oocyte harvest. PMID- 11491378 TI - Morphological diagnosis of HPV lesions and cervical intraepithelial neoplasia (CIN) is highly reproducible. AB - PURPOSE: to assess the value of individual histological criteria in the diagnosis of cervical HPV lesions. METHODS: 138 women referred for colposcopic evaluation (due to abnormal PAP smears) were subjected to cervical punch biopsy. The biopsies were classified as no HPV lesion, CIN 1, or CIN 2-3 by two observers independently. Kappa tests were used for interobserver agreement of the diagnosis. The presence of binucleation, multinucleation, abnormal mitosis. koilocytosis, spindle koilocytosis and dyskeratosis was similarly assessed. RESULTS: the Kappa statistic was 0.638 (CI 95% 0.533-0.743), showing substantial inter-observer agreement. Abnormal mitosis and multi-nucleation were the two most powerful discriminators between CIN 2-3 and CIN 1. Koilocytosis proved to be the single most powerful discriminator between CIN 1 lesions and non-HPV lesions. CONCLUSION: the results advocate the use of histology as the gold standard in diagnosing cervical precancerous lesions. The classical criteria can be also used to differentiate low-grade lesions, which has practical implications by avoiding the unnecessary treatment of minor abnormalities. PMID- 11491379 TI - Beneficial effects of low doses of ethinyl-estradiol on the lipid profile in postmenopausal women. AB - The purpose of this study was to investigate the beneficial effects of low doses of ethinyl-estradiol on the lipid profile in postmenopausal women. One hundred and five patients (mean age [+/-S D] 42.9 +/- 5.0 years) who underwent a hysterectomy and bilateral salpingo-oophorectomy were included in the study. For the present study serum levels of total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, apolipoprotein B (apoB), and lipoprotein(a) [Lp(a)] were investigated. When all patients were considered together (Table 1), EE2 therapy significantly increased serum levels of total cholesterol, HDL cholesterol and LDL cholesterol. The ratio of HDL to LDL cholesterol, Lp(a) and triglyceride concentrations did not change significantly from the baseline value. Although our study was not randomized or controlled with a placebo, the beneficial metabolic effects of ethinyl-estradiol on lipid patterns should be considered in patients needing hormonal replacement therapy in postmenopause. PMID- 11491380 TI - Ovarian cancer in a woman previously diagnosed with endometriosis and an extremely high serum CA-125 level. AB - PURPOSE: Follow-up of a woman with a serum CA-125 level > 1000 U/mL where laparoscopy only found endometriosis. METHODS: Case report-- re-evaluation several years later. RESULTS: Extensive clear-cell carcinoma of ovary with metastases leading to death. CONCLUSION: This case suggests that bilateral oophorectomy should be performed in women not desiring any more children if the serum CA- 125 level is very high even if only endometriosis is found initially. PMID- 11491381 TI - Effect of fibroids on cumulative probability of pregnancy in women taking follicle maturing drugs without assisted reproductive technology. AB - PURPOSE: To determine if the presence of uterine fibroids adversely affect in vivo conception rates. METHODS: Pelvic ultrasound evaluation of the presence or absence of fibroids in consecutive infertility patients not treated with assisted reproductive technology in a two-year period. Conception outcome noted. Data analyzed according to the presence or absence of fibroids. RESULTS: No difference in cumulative probability of pregnancy after five months was seen in patients with or without the presence of fibroids. There were no confounding variables found that could have skewed the pregnancy rates in one direction or the other. CONCLUSION: In general the presence of fibroids do not adversely affect conception outcome for in vivo pregnancies. However, since the majority of the fibroids were small (< 6 cm) and were not submucosal and did not compress the endometrial cavity, larger studies are needed to address specific subtypes and circumstances on pregnancy outcome. PMID- 11491382 TI - Successful treatment of cervical and simultaneous cervico-isthmic pregnancy with methotrexate. AB - OBJECTIVE: To describe the monitoring of a case of cervical and simultaneous cervico-isthmic pregnancy. SETTING: University of Bari (Italy), Department of Obstetrics and Gynecology. PATIENT: A 30-year-old white woman, nulliparous, at 8 weeks of amenorrhoea. INTERVENTION: Methotrexate and folinic acid administered systemically. MAIN OUTCOME MEASURE: Treatment success was defined as elimination of the cervical and cervico-isthmic pregnancy, with non-invasive treatment and preservation of the uterus and normal ovarian activity restored. RESULT: Methotrexate and folinic acid were administered, elimination of a twin pregnancy with declining serum beta-hCG levels and with ultrasound was observed. The patient had only occasional dark vaginal bleeding and temporary movement of the transaminase. CONCLUSION: This case report shows that methotrexate is a valid, conservative and non-invasive treatment for a patient affected by cervical pregnancy who wishes to keep fertility. PMID- 11491383 TI - A successful pregnancy from zygotes cryopreserved for >9 years: case report. AB - PURPOSE: To evaluate the longest interval that embryos can remain frozen and still result in a viable pregnancy after thaw and transfer. METHODS: Case report. RESULTS: A 42-year-old woman conceived and successfully completed the first trimester after transfer of frozen donated embryos from a 32-year-old woman whose eggs had been cryopreserved for over nine years. CONCLUSIONS: Patients can now be informed that there is anecdotal evidence that embryos can survive and result in successful pregnancies even after nine years of cryopreservation. This information may affect legislative decisions in requiring destruction of these stored embryos. PMID- 11491384 TI - Quantitative evaluation of collagen and muscle fibers in the lower urinary tract of castrated and under-hormone replacement female rats. AB - OBJECTIVE: To evaluate the number of collagen and muscle fibers in the muscle layer of the urethra and in the bladder wall of castrated and under-hormone replacement female rats. METHOD: We studied 37 castrated female rats assigned to the following groups: Group C (n=9): received no medication; Group P (n=8) was given 0.1 ml of placebo, subcutaneous (SC) route; Group E (n=10): 17beta estradiol, 10 microg/kg/day, SC route; Group PR (n=9): medroxyprogesterone acetate. 0.2 mg/kg/day, SC route; Group E+PR (n=9): association of 17beta estradiol and medroxyprogesterone acetate. Sections were taken from the bladder wall and from the middle third of the urethra, and the specimens were stained with picrosirius for collagen and muscle fiber identification. RESULTS: Groups C and P showed a similar amount of collagen in the bladder and in the urethra, however greater than the other groups. Group E showed the smallest number of collagen fibers in the urethra. Groups E and E+PR presented a larger number of muscle fibers in the bladder. Group PR presented a larger number of muscle fibers than groups C and P, however smaller than groups E and E+PR. In the muscle layer of the urethra, the number of collagen fibers was smaller in Group E than in all the other groups, which were similar among one another. In regard to the urethral muscles, Group E was found to present the largest number of muscle fibers as compared to the other groups analyzed, while Group PR showed a significant decrease in the muscle layer, even in relation to the groups that were given no hormone medication. CONCLUSION: Estrogens significantly decrease the amount of collagen fibers, increase the amount of muscle fibers and determine a significantly decreased collagen/muscle ratio in both the detrusor muscle and in the urethral muscle layer. It is also noticed that isolated progestogen decreases the amount of collagen fibers and increases the number of muscle fibers in the detrusor muscle, but with less intensity than replacement with estrogens alone. It neither alters the number of collagen fibers nor decreases the muscle fibers in the muscle layer of the urethra, with increased collagen/muscle ratio in that structure. Finally, the estrogen-progestogen combination determines significantly decreased collagen fibers and increased muscle fibers in the detrusor muscle, causing no alteration to the collagen or muscle fibers in the muscle layer of the urethra. PMID- 11491385 TI - Endometriosis arises from implant of endometriotic cells outside the uterus: a report of active vesicouterine centrifugal fistula. AB - Vesicouterine fistula is rare, accounting for nearly 4% of all urogenital fistulas. Lower segment cesarean delivery is the main predisposing event but in the last few years other possible predisposing factors have been pointed out. Clinically, it can show itself in different forms and the diagnosis is often delayed although it is not difficult. We report our experience about a case of postcesarean vesicouterine fistula arising on a focus of vesical endometriosis and we discuss an eventual hypothetical pathogenetic correlation between bladder endometriosis and uterovesical fistula. PMID- 11491386 TI - Metabolism of amiodarone (Part III): identification of rabbit cytochrome P450 isoforms involved in the hydroxylation of mono-N-desethylamiodarone. AB - 1. Amiodarone (AMI) is a potent anti-arrhythmic drug and mono-N desethylamiodarone (MDEA) is its only known metabolite. It was found recently that in rabbit liver microsomes MDEA was biotransformed to n-3-hydroxybutyl-MDEA (3OH-MDEA). 2. In liver microsomes isolated from the untreated rabbit, the formation of 3OH-MDEA obeyed Michaelis-Menten enzyme kinetics with Km = 6.39 +/- 1.07 microM and Vmax = 0.56 +/- 0.21 nmolmin(-1) mg(-1) protein. 3. Furthermore, (1) among chemicals usually used as inhibitors of cytochrome P450, only midazolam (MDZ), cyclosporin A and ketoconazole inhibited the MDEA hydroxylase activity significantly (>60% inhibition), (2) MDZ, a substrate of CYP3A, inhibited the 30OH-MDEA formation competitively (Ki = 10 +/- 5 microM), (3) the formation rates of 3OH-MDEA correlated positively with those of 1'OH-MDZ (r = 0.81; n = 6), and (4) MDEA hydroxylase activity of microsomes isolated from rabbit rifampicin induced cultured hepatocytes was 4-fold more active than the control. 4. Since CYP3A6 is mainly induced by rifampicin in rabbit-cultured hepatocytes, the data suggest that this isoform is involved in the biotransformation of MDEA to 3OH MDEA. 5. Since alpha-naphthoflavone, cimetidine and quinidine also partially inhibited the MDEA hydroxylase activity, it is possible that other CYPs, such as 1A, 2C and 2D, may also be active in the metabolism of amiodarone. PMID- 11491387 TI - Role of endogenous nitric oxide in liver-specific functions and survival of cultured rat hepatocytes. AB - 1. The role of endogenous nitric oxide in rat hepatocyte functionality and survival in cell culture was examined. Towards this aim, cytochrome P450 activities (CYP1A1/2, 2B1, 2A1, 2C11, 2D1, 2E1 and 3A1), liver-specific metabolic functions and cell survival were comparatively evaluated in hepatocytes isolated from the male Sprague-Dawley rat and/or cultured in control conditions or in the presence of N-omega-nitro-L-arginine methyl ester (NAME), an inhibitor of nitric oxide synthesis. 2. Suppression of nitric oxide production by NAME paralleled a substantial preservation of hepatocyte phenotype in culture. The presence of NAME was particularly important during isolation and/or the 6-24h culture. By 24h, beneficial effects were evident in parameters particularly unstable in culture (glycogen content, P450), whereas no changes were produced in well-preserved functions (glucose, urea and albumin synthesis, glutathione, drug-conjugating enzymes). 3. Long-term treatment of hepatocytes with NAME also produced a reduction in caspase 3 activation and in the percentage of spontaneous apoptotic cells, and an increase in cell survival and transcriptional activity as shown by attached cellular protein content and the protein-DNA ratio respectively. 4. In conclusion, inhibition of early endogenous nitric oxide formation is an efficient procedure for obtaining hepatocyte cultures with stable expression of differentiated functions, high cell survival and few signs of cell senescence. PMID- 11491388 TI - Cooperativity of alpha-naphthoflavone in cytochrome P450 3A-dependent drug oxidation activities in hepatic and intestinal microsomes from mouse and human. AB - 1. The effects of several CYP3A substrates (alpha-naphthoflavone (alphaNF), terfenadine, midazolam, erythromycin) on nifedipine oxidation and testosterone 6beta-hydroxylation activities were investigated in hepatic and intestinal microsomes from mouse and human. 2. alphaNF (10 microM) and terfenadine (100 microM) inhibited nifedipine oxidation activities (at substrate concentration of 100 microM) in mouse hepatic microsomes to approximately 50%, but not in mouse intestinal microsomes. alphaNF (30 microM) stimulated nifedipine oxidation activities in mouse and human intestinal microsomes and in human hepatic microsomes to approximately 1.3-1.8-fold. Inhibitory potencies (50% inhibition concentration, IC50) of midazolam and erythromycin for nifedipine oxidations were calculated to be approximately 90 microM in human intestinal microsomes. In contrast, testosterone (100 microM) stimulated the nifedipine oxidation activities approximately 1.5-fold in hepatic and intestinal microsomes from mouse and human. 3. alphaNF showed different effects on the kinetic parameters including the Hill coefficients of nifedipine oxidation and testosterone 6beta hydroxylation catalysed by hepatic and intestinal microsomes from mouse and human. Cooperativity in nifedipine oxidation was increased by the addition of alphaNF to pooled human hepatic microsomes, but little effects of alphaNF could be observed in individual human intestinal microsomes. 4. These results suggest that CYP3A enzymes in liver and intestine might have different characteristics and that observations from hepatic microsomes should not be directly applicable to intestine metabolism in some cases. Studies of drug-drug interactions of CYP3A substrates are recommended to be performed using intestinal samples. PMID- 11491389 TI - Identification and reactivity of the major metabolite (beta-1-glucuronide) of the anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) in humans. AB - 1. The novel anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) is extensively metabolized by glucuronidation and 6-methylhydroxylation, resulting in DMXAA acyl glucuronide (DMXAA-G) and 6-hydroxymethyl-5-methylxanthenone-4 acetic acid (6-OH-MXAA). 2. The major human urinary metabolite of DMXAA was isolated and purified by a solid-phase extraction (SPE) method. The isolated metabolite was hydrolysed to free DMXAA by strong base, and by beta glucuronidase. Liquid chromatography-mass spectrometry (LC-MS) and spectral data indicated the presence of a molecular ion [M + 1]+ at m/z 459, which was consistent with the molecular weight of protonated DMXAA-G. 3. The glucuronide was unstable in buffer at physiological pH, plasma and blood with species variability in half-life. Hydrolysis and intramolecular migration were major degradation pathways. 4. In vitro and in vivo formation of DMXAA-protein adducts was observed. The formation of DMXAA-protein adducts in cancer patients receiving DMXAA was significantly correlated with plasma DMXAA-G concentration and maximum plasma DMXAA concentration. 5. At least five metabolites of DMXAA were observed in patient urine, with up to 60% of the total dose excreted as DMXAA-G, 5.5% as 6 OH-MXAA and 4.5% as the glucuronide of 6-OH-MXAA. 6. These data suggest that the major metabolite in patients' urine is DMXAA beta-1-glucuronide, which may undergo hydrolysis, molecular rearrangement and covalent binding to plasma protein. The reactive properties of DMXAA-G may have important implications for the pharmacokinetics, pharmacodynamics and toxicity of DMXAA. PMID- 11491390 TI - Glutathione depletion in rat hepatocytes: a mixture toxicity study with alpha, beta-unsaturated esters. AB - 1. Glutathione (GSH) depletion is often reported as an early cytotoxic effect, caused by many reactive organic chemicals. In the present study, GSH depletion in primary rat hepatocytes was used as an in vitro effect-equivalent to measure the toxic potency of alpha,beta-unsaturated esters (acrylates and methacrylates). 2. When these compounds were administered as a mixture, GSH depletion was dose additive. The result of the mixture study shows that GSH depletion may be a useful effect-equivalent for the risk assessment of mixtures of alpha,beta unsaturated esters. 3. To get more insight in the underlying mechanisms of GSH depletion, the metabolism of two esters was investigated in greater detail. One of them, allyl methacrylate, was metabolized to acrolein. This metabolic pathway can explain the high potency of allyl methacrylate to deplete GSH despite its low intrinsic chemical reactivity. PMID- 11491391 TI - Four new stilbene dimers from the lianas of Gnetum hainanense. AB - Four new stilbene dimers, gnetuhainins P (1), Q (2), K (3) and L (4), were isolated from the lianas of Gnetum hainanense C. Y. Cheng. Their structures and relative configurations were determined on the basis of spectroscopic evidence, especially 2D NMR techniques. PMID- 11491392 TI - Atemoyacin E, a bis-tetrahydrofuran annonaceous acetogenin from Annona atemoya seeds. AB - Atemoyacin E (I), a new adjacent bis-tetrahydrofuran annonaceous acetogenin was isolated and characterized from the seeds of Annona atemoya. PMID- 11491393 TI - Chinese bittersweet alkaloid III, a new compound from Celastrus angulatus. AB - A novel skeleton alkaloid named Chinese Bittersweet Alkaloid III from the leaves of C. angulatus was reported. The structure and its stereochemistry were established by IR, ID and 2DNMR (1H-1HCOSY, HMQC, HMBC, NOESY), MS and elemental analysis. PMID- 11491395 TI - The structure of physalin T from Physalis alkekengi var. franchetti. AB - A new steroidal constituent named physalin T (3) was isolated from the aqueous extract of Physalis alkekengi var. francheti. Based on 1H and 13C NMR spectral studies the structure was assigned as 2,3-dihydrophysalin D, i.e., 5alpha,6beta dihydroxy-2,3,5,6-tetrahydrophysalin B, which is the first example of a natural physalin possessing a saturated ring A moiety. The structure was confirmed by the chemical transformation from the known physalin D (2) to physalin T. PMID- 11491394 TI - Sesquiterpene glucosides from anti-leukotriene B4 release fraction of Taraxacum officinale. AB - Chemical examination of the MeOH extract of the root of Taraxacum officinale, which exhibited inhibitory activity on the formation of leukotriene B4 from activated human neutrophils, has resulted in the isolation of 14-O-beta-D glucosyl-11,13-dihydro-taraxinic acid (1) and 14-O-beta-D-glucosyl-taraxinic acid (2). The absolute stereostructure of 1 has been established by X-ray chrystallographic examination. PMID- 11491396 TI - Triterpene glycoside from Terminalia arjuna. AB - A new triterpene glycoside, arjunetoside, together with oleanolic and arjunic acids has been isolated from the root bark of Terminalia arjuna. The structure of arjunetoside has been established as 3-O-beta-D-glucopyranosyl-2alpha,3beta, 19alpha-trihydroxyolean-12-en-28-oic acid, 28-O-beta-D-glucopyranoside by chemical and spectral data. PMID- 11491397 TI - Eclalbatin, a triterpene saponin from Eclipta alba. AB - From the whole plant of Eclipta alba, a new triterpene saponin, named eclalbatin, together with alpha-amyrin, ursolic acid and oleanolic acid were isolated. The structure of eclalbatin has been established as 3-O-beta-D-glucopyranosyl-3-beta hydroxy-olean-12-en-28-oic acid, 28-O-beta-D-arabinopyranoside (1) on the basis of chemical and spectral data. PMID- 11491398 TI - Synthesis and biological activity of amide derivatives of ginkgolide A. AB - Amide derivatives of ginkgolide A were prepared and evaluated for their in vitro ability to inhibit the PAF-induced aggregation of rabbit platelets. They showed less activities than their parent compound ginkgolide A. PMID- 11491399 TI - A new phenolic glycoside and a new trans-clerodane diterpene from Conyza blinii. AB - A new phenolic glycoside, 4-propionyl-2,6-dimethoxyphenyl beta-D-glucopyranoside (1) and a new trans-clerodane diterpene named 19-deacetylconyzalactone (2), were isolated from the aerial parts of Conyza blinii. PMID- 11491400 TI - A pair of 24-hydroperoxyl epimeric dammarane saponins from flower-buds of Panax ginseng. AB - Further investigation on the saponins of the flower-buds of Panax ginseng C. A. Meyer has resulted in the isolation and structural elucidation of a pair of new 24-epimers of dammarane type saponins named ginsenoside I and II. The structures of the epimers were characterized on the basis of chemical and spectral evidence as 3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20-S-O-beta-D glucopyranosyl-3beta, 12beta,20(S)-trihydroxy-24xi-hydroperoxydammar-25-ene, except for their C-24 configurations. Ginsenoside I is a new triterpene glycoside, and ginsenoside II is a known compound first isolated from a natural plant. PMID- 11491402 TI - Cryopreservation of epididymal spermatozoa collected by needle biopsy from cynomolgus monkeys (Macaca fascicularis). AB - We have examined the motility, morphology, and cryopreservation of epididymal spermatozoa collected by needle biopsy from cynomolgus monkeys (Macaca fascicularis). At collection, epididymal sperm (23 x 10(6) +/- 4 x 10(6) sperm/sample; 611 x 10(6) +/- 116 x 10(6) sperm/ ml; n = 18) were alive (79 +/- 2%), motile (67 +/- 2%), and exhibited intact membranes (65 +/- 2%). Sperm maintained at room temperature in handling medium exhibited decreased motility over time, but head-to-head agglutination was limited. Tris egg-yolk extender containing 6% glycerol and dimethylsulfoxide (DMSO) did not significantly affect functional morphology, whereas extender containing propanediol significantly reduced motility, survival, and membrane integrity. Cryostorage reduced all measures of functional morphology independent of cryoprotectant. Post-thaw motility was superior for glycerol and DMSO compared to propanediol. Variation in glycerol concentration (4, 6, and 8%) produced equivocal effects on sperm functional morphology post-thaw. Needle biopsy may be a useful technique for laboratory and field-based collection of spermatozoa from nonhuman primates. PMID- 11491401 TI - A cyclic peroxide of clerodenoic acid from the Taiwanese liverwort Schistochila acuminata. AB - Two minor diterpenoids, 15(16)-peroxy-3, 13-clerodadien-18-oic acid (4) and totarol (7), were isolated from the liverwort Schistochila acuminata. The former is a further oxidized cyclic peroxide from the major component. The latter is a rearranged abietane alcohol which skeleton was found in liverworts for the first time. PMID- 11491403 TI - Immunolocalization of proliferating cell nuclear antigen (PCNA) in cynomolgus monkey (Macaca fascicularis) testes during postnatal development. AB - The age-related distribution of proliferating cell nuclear antigen (PCNA) in the testes of cynomolgus monkeys (Macaca fascicularis) during postnatal development was detected using light-microscopic immunohistochemistry. In neonatal testes, some PCNA-positive spermatogonia, Sertoli cells, peritubular cells, and Leydig cells were detected. In early infantile testes, only a few of these cell types were positive. In late infantile testes, the numbers of positive cells were greater than in the earlier developmental stages. In pubertal testes, the numbers of positive spermatogonia, spermatocytes, Sertoli cells, peritubular cells, and Leydig cells were considerably higher. In adult testes, a larger percentage of spermatogonia and spermatocytes was positive, and peritubular cells and Leydig cells were occasionally positive; secondary spermatocytes, spermatids, and Sertoli cells were not positive. We concluded that immunolocalization of PCNA can serve as a tool for studying proliferation status in developing testes of cynomolgus monkeys. A relatively low proliferative activity in early infantile testes and a remarkable increase of proliferative activity in pubertal testes correlate with the fluctuations of steroidogenic functions during postnatal development in cynomolgus monkeys. PMID- 11491405 TI - A spontaneously occuring mammary gland ductal carcinoma in situ in a rhesus macaque (Macaca mulatta) and a review of spontaneous mammary gland tumors in rhesus monkeys. AB - A spontaneous mammary gland ductal carcinoma in situ was diagnosed in a 6-8-year old female rhesus macaque (Macaca mulatta). To our knowledge, this is only the tenth case of spontaneous mammary gland tumors to be reported in rhesus monkeys. Despite the paucity of case reports, several theories exist to explain the occurrence of mammary tumors. The Mason Pfizer monkey virus, a type D retrovirus similar to the virus that causes simian acquired immunodeficiency syndrome, has been implicated as a possible etiologic agent. Because this virus has been isolated from normal primate mammary tissue, it is unlikely to be the sole etiologic agent. Other theories include the tumorogenic effects that androgens, growth hormones, irradiation, and aging have on the mammary gland. PMID- 11491404 TI - Seasonal changes of humoral and cellular immune responses to Japanese cedar (Cryptomeria japonica) pollen allergens in Japanese monkeys (Macaca fuscata) with pollinosis. AB - The natural occurrence of Japanese cedar [Cryptomeria japonica (CJ)] pollinosis has been reported in Japanese monkeys (Macaca fuscata). The present study was designed to investigate seasonal changes in immunological reactions to CJ pollen allergens in monkeys with CJ pollinosis. Blood samples were collected from six monkeys with CJ pollinosis before and after CJ pollen season. Seasonal changes in specific IgE and IgG to major allergens (Cry j 1 and Cry j 2) were observed before and after CJ pollen season. The humoral responses decreased significantly before CJ pollen and increased after CJ pollen season. Similar seasonal changes in peripheral blood mononuclear cells proliferative responses to CJ allergens were observed before and after CJ pollen season. These humoral and cellular immune responses might serve as a biomarker for assessing new immunotherapies for monkeys with pollinosis. PMID- 11491406 TI - Subcutaneous hemangiosarcomas in a rhesus macaque (Macaca mulatta). AB - Hemangiosarcoma is a malignant tumor of vascular endothelial cell origin. The occurrence of hemangiosarcoma in nonhuman primates has been rarely documented. An adult male rhesus monkey was reported having a firm subcutaneous swelling, approximately 4.5 cm in diameter, on the ventral midline of the abdomen. Fine needle aspiration, microbial culture, biopsy, radiographs, exploratory laparotomy, histopathology, immunohistochemistry, hematology, and serology were performed. A second subcutaneous mass approximately 4.5 x 4.0 x 2.7 cm developed on the ventral midline several weeks later. A fine-needle aspirate of the first mass consisted of numerous erythrocytes with few polymorphonuclear cells and lymphocytes. Histopathology showed foci of spindle-shaped cells surrounding the vascular spaces. Many spindle-shaped cells had prominent nucleoli, and mitotic figures could occasionally be seen. Immunohistochemical staining of the masses for Factor VIII-related antigen, an endothelial cell and tumor marker, yielded positive results. Both masses were consistent with hemangiosarcoma. PMID- 11491407 TI - The rehabilitation of captive baboons. AB - Eleven baboons who had been singly housed indoors for an average of 5 years were moved to outdoor social groups in an attempt to provide a more species-typical environment and reduce high levels of abnormal behavior. Nine of the baboons were observed while in single housing and, over a 6-month period, while housed outdoors socially to document long-term changes in behavior. Abnormal behavior decreased significantly from an average of 14% of the observation time in the single cages to 3% in the sixth month of social housing. Cage manipulation and self-directed behaviors also significantly decreased, while social behavior, enrichment-directed behavior, and locomotion increased in social housing. Baboons that had been in long-term indoor single housing were able to reproduce and form stable social groups without injury. This study provides evidence that even behaviorally disturbed nonhuman primates can be successfully rehabilitated to live in social groups. PMID- 11491408 TI - Radiation-induced morphologic changes in the rhesus monkey (Macaca mulatta) brain. AB - The right cerebral hemisphere of 24 rhesus monkeys scheduled for necropsy at the completion of another project were studied histopathologically 1-30 days after a single dose of 60Co-irradiation. Histopathologically, inflammation and gliosis consistently occurred at specific time points but varied in severity between individuals. Multifocal hemorrhage, edema, and an acute neutrophilic inflammatory response were observed initially whereas perivascular accumulations of lymphocytes were observed in specimens at the end of the study. Microglia/macrophages were most prominent during the first week after irradiation, whereas astrocytes were reactive throughout the observation period. The early clinical manifestations of the central nervous system (CNS), because of brain irradiation in humans, correspond temporally with acute vascular responses, acute and subacute inflammatory cell responses, and subacute demyelination and reactive astrocytic and microglial responses observed in the rhesus monkey. Initial responses of the CNS to gamma-irradiation may have potential implications for the development of radiation-induced late injury of the CNS. PMID- 11491409 TI - Ultrasound detection of non-Hodgkin's lymphoma in three cynomolgus monkeys after renal transplantation and cyclosporine immunosuppression. AB - PURPOSE: To describe the early detection of non-Hodgkin's lymphoma (NHL) with ultrasound in three clinically normal cynomolgus monkeys post-renal transplantation and immunosuppression with cyclosporine. MATERIALS AND METHODS: The monkeys in this report were treated with cyclosporine (Neoral) after receiving renal transplants. In addition to clinical and laboratory (hematology, serum chemistry) monitoring, renal allografts were monitored every 2 weeks with ultrasound and ultrasound-guided allograft biopsies were performed. RESULTS: Enlarged renal hilar and mesenteric lymph nodes were detected with ultrasound in three monkeys on days 36, 49 and 134 post-transplantation. Sonographically the lymph nodes were inhomogeneous, of low echogenity and rounded. In two animals, the spleen was sonographically enlarged and inhomogeneous. All three monkeys were symptom-free at the time of ultrasound detection and NHL was diagnosed histologically. CONCLUSION: Ultrasound provides a rapid, non-invasive means of early detection of NHL in animal transplantation models prior to the onset of clinical symptoms of disease. PMID- 11491410 TI - Accuracy and precision of dual-energy X-ray absorptiometry for body composition measurements in rhesus monkeys. AB - Accuracy of body composition measurements by dual-energy X-ray absorptiometry (DXA) was compared with direct chemical analysis in 10 adult rhesus monkeys. DXA was highly correlated (r-values > 0.95) with direct analyses of body fat mass (FM), lean mass (LM) and lumbar spine bone mineral content (BMC). DXA measurements of total body BMC were not as strongly correlated (r-value = 0.58) with total carcass ash content. DXA measurements of body FM, LM and lumbar spine BMC were not different from data obtained by direct analyses (P-values > 0.30). In contrast, DXA determinations of total BMC (TBMC) averaged 15%, less than total carcass ash measurements (P = 0.002). In conclusion, this study confirms the accurate measurement of fat and lean tissue mass by DXA in rhesus monkeys. DXA also accurately measured lumbar spine BMC but underestimated total body BMC as compared with carcass ash determinations. PMID- 11491411 TI - Meta-analysis, overviews and publication bias. PMID- 11491412 TI - A sensitivity analysis for publication bias in systematic reviews. AB - There is no simple method of correcting for publication bias in systematic reviews. We suggest a sensitivity analysis in which different patterns of selection bias can be tested against the fit to the funnel plot. Publication bias leads to lower values, and greater uncertainty, in treatment effect estimates. Two examples are discussed. An appendix lists the S-plus code needed for carrying out the analysis. PMID- 11491413 TI - Approximations for trimmed Fisher procedures in research synthesis. AB - When combining the results of independent studies it often happens that some studies are potentially aberrant either in quality or in actual values. Because aberrant studies are often at the extremes, we may wish to trim some of the largest and smallest effects. In such a case the use of p-values may well serve as a diagnostic method. However, the use of ordered effects changes the distribution of the underlying statistics. We provide a discussion of the exact distribution of the trimmed version of the Fisher procedure. Because of the complexity of the exact distribution, several approximations are presented. These alternatives are applied to a meta-analysis on the effect of the dose of a drug on the risk of mortality. PMID- 11491414 TI - Bayesian methods in meta-analysis and evidence synthesis. AB - This paper reviews the use of Bayesian methods in meta-analysis. Whilst there has been an explosion in the use of meta-analysis over the last few years, driven mainly by the move towards evidence-based healthcare, so too Bayesian methods are being used increasingly within medical statistics. Whilst in many meta-analysis settings the Bayesian models used mirror those previously adopted in a frequentist formulation, there are a number of specific advantages conferred by the Bayesian approach. These include: full allowance for all parameter uncertainty in the model, the ability to include other pertinent information that would otherwise be excluded, and the ability to extend the models to accommodate more complex, but frequently occurring, scenarios. The Bayesian methods discussed are illustrated by means of a meta-analysis examining the evidence relating to electronic fetal heart rate monitoring and perinatal mortality in which evidence is available from a variety of sources. PMID- 11491415 TI - Antiretroviral activity and safety of abacavir in combination with selected HIV-1 protease inhibitors in therapy-naive HIV-1-infected adults. AB - OBJECTIVE: To assess antiretroviral efficacy and safety of abacavir in combination with selected HIV-1 protease inhibitors. DESIGN: A 48-week, open label study. MATERIALS AND METHODS: Eighty-two antiretroviral naive HIV-1 infected adults (CD4 cell count > or = 100 cells/mm3, plasma HIV-1 RNA > or = 5,000 copies/ml) were randomly assigned to receive abacavir (300 mg twice daily) in combination with standard doses of one of five protease inhibitors: indinavir, saquinavir soft-gel, ritonavir, nelfinavir or amprenavir. Adults who met protocol defined switch criteria at or after week 8 could modify their randomized therapy. Antiretroviral activity was assessed by the proportion of subjects with plasma HIV-1 RNA < or = 400 and < or = 50 copies/ml, and by changes in plasma HIV-1 RNA levels and CD4 cell counts. Safety was assessed by monitoring clinical adverse events and laboratory abnormalities. RESULTS: At week 48, the proportion of subjects in the indinavir, saquinavir, ritonavir, nelfinavir and amprenavir groups with plasma HIV-1 RNA < or = 400 copies/ml was 53, 50, 50, 41 and 56%, respectively, and the proportion with HIV-1 RNA < or = 50 copies/ml was 47, 56, 50, 47, and 44%, respectively (by intent-to-treat analysis). Median reductions from baseline in plasma HIV-1 RNA for each group ranged from 1.7 to 2.4 log10 copies/ml. The median CD4 cell count increase from baseline was 195, 131, 116, 136 and 259 cells/mm3 in the indinavir, saquinavir, ritonavir, nelfinavir, and amprenavir groups, respectively. Overall, the most common adverse events attributed to study drugs were diarrhoea, nausea, malaise/fatigue, headache and perioral paresthesia. The frequency of treatment-limiting adverse events did not differ between groups. CONCLUSIONS: Abacavir is safe and effective when used in combination with a protease inhibitor. PMID- 11491416 TI - Increased drug susceptibility of HIV-1 reverse transcriptase mutants containing M184V and zidovudine-associated mutations: analysis of enzyme processivity, chain terminator removal and viral replication. AB - The presence of the HIV reverse transcriptase (RT) resistance mutation, M184V, induced by lamivudine and abacavir treatment results in increased tenofovir, adefovir and zidovudine susceptibility for HIV-1 with zidovudine-associated RT mutations in vitro. Treatment with oral prodrugs of tenofovir and adefovir has resulted in substantial HIV-1 RNA reductions in antiretroviral-experienced patient populations who have lamivudine- and zidovudine-resistant HIV-1. An enzymatic analysis was undertaken to elucidate the mechanisms of altered drug susceptibilities of HIV-1 containing zidovudine-associated mutations in the presence or absence of M184V. The inhibition constants (Ki) for the active metabolites of tenofovir, adefovir and zidovudine did not vary significantly between recombinant mutant and wild-type RT enzymes. Although increased removal of chain-terminating inhibitors by pyrophosphorolysis and ATP-dependent unblocking correlated with reduced susceptibility of viruses with zidovudine associated mutations, a reduction in the removal of chain-terminators was not observed, which would explain the increased drug susceptibility of mutants containing M184V plus zidovudine-associated mutations. However, analyses of single-cycle processivity of the mutant RT enzymes on heteropolymeric RNA templates showed that all M184V-containing mutant RT enzymes were less processive than wild-type RT, most notably for mutants expressing both zidovudine-associated mutations and M184V. Similarly, the in vitro replication capacity of a mutant virus expressing a zidovudine-associated mutation and M184V was significantly reduced compared with wild-type virus. The observed decrease in enzymatic processivity of the M184V-expressing RT enzymes might result in decreased viral replication, which then might contribute to the increased drug susceptibility of HIV-1 expressing these RT mutations. PMID- 11491417 TI - AVANTI 3: a randomized, double-blind trial to compare the efficacy and safety of lamivudine plus zidovudine versus lamivudine plus zidovudine plus nelfinavir in HIV-1-infected antiretroviral-naive patients. AB - The objective of our randomized, multicentre, double-blind, placebo-controlled study was to investigate the safety, tolerability, and antiretroviral and immunological effect of double and triple combination therapy regimens. A total of 105 antiretroviral therapy-naive patients were randomized to receive either zidovudine (300 mg twice per day) plus lamivudine (150 mg twice per day) plus nelfinavir placebo (three times per day) (n=52), or zidovudine/lamivudine (dose as before) plus nelfinavir (750 mg three times per day) (n=53) for 28 weeks. After this time, patients were allowed to switch to open-label zidovudine/lamivudine/nelfinavir. The overall log10 reduction from baseline in plasma HIV-1 RNA was significantly greater in the zidovudine/lamivudine/nelfinavir group than the zidovudine/lamivudine group (P=0.001; median treatment difference, -1.01 log10 copies/ml; 95% confidence interval -1.23 to -0.79), as measured by the average area under the curve minus baseline over 28 weeks. Increases from baseline in CD4 cell counts were statistically significantly greater in the zidovudine/ lamivudine/nelfinavir group (101.5 cells/ml) than the zidovudine/lamivudine group (47.0 cells/ml; P=0.027) at week 28. Of note, the addition of nelfinavir from weeks 28-52 led to an increase in the proportion of subjects with plasma HIV-1 RNA <400 copies/ml from 17% (9/52 patients on zidovudine/lamivudine) to 50% (13/26 patients who switched to zidovudine/lamivudine/nelfinavir). Incidence of drug-related adverse events was similar in the two groups, except for nausea (more common in zidovudine/lamivudine group; 40 versus 17%) and diarrhoea (more common in zidovudine/lamivudine/nelfinavir group; 45 versus 14%). In conclusion, our study confirms the efficacy of triple combination therapy with two nucleoside analogues and a protease inhibitor compared with double-nucleoside therapy. Interestingly, the addition of nelfinavir to zidovudine/lamivudine, even after 6 months of double nucleoside therapy, led to a substantial virological benefit that was sustained over 24weeks in a subset of patients. PMID- 11491418 TI - Treatment intensification with abacavir in HIV-infected patients with at least 12 weeks previous lamivudine/zidovudine treatment. AB - OBJECTIVES: To demonstrate that lamivudine and zidovudine, given separately (lamivudine/zidovudine) or as a single combination tablet (Combivir), had equivalent efficacy. To evaluate the safety and antiretroviral activity of intensification with abacavir in patients treated with lamivudine/zidovudine for > or = 12 weeks. DESIGN: A 12-week, equivalence study of lamivudine/ zidovudine versus Combivir. Patients who completed this study could enter a 48-week, intensification study of Combivir plus abacavir. METHODS: In the equivalence study, treatment-naive patients were assessed for HIV-1 RNA, CD4 cell count and genotype. The same assessments plus phenotype were made in the intensification study. Serious adverse events were recorded in the equivalence study and all adverse events in the intensification study. RESULTS: Lamivudine/zidovudine (n=40) and Combivir (n=35) gave equivalent reductions in plasma HIV-1 RNA levels at week 12. An identical proportion of patients (74%) in each treatment group harboured virus with the M184V mutation after 12 weeks. Fifty-two patients entered the intensification study and 44 completed 48 weeks of treatment. At the time of intensification with abacavir, all 35 patients with evaluable isolates harboured HIV-1 containing M184V. Addition of abacavir to Combivir led to further decreases in plasma HIV-1 RNA and increases in CD4 cell counts compared with the start of intensification (P<0.001 at week 48). After 48 weeks of triple therapy, multi-nucleoside resistance mutations at codons 69 and 151 were not detected in any patients. All treatment regimens were generally well tolerated. CONCLUSION: Lamivudine/zidovudine and Combivir have equivalent antiretroviral activity over 12 weeks. Adding abacavir to Combivir can be a safe and effective therapeutic option for patients, including those harbouring virus with the M184V mutation. PMID- 11491419 TI - In vitro anti-HIV-1 synergy between non-nucleoside reverse transcriptase inhibitors nevirapine and efavirenz. PMID- 11491420 TI - Antiretroviral activity of emtricitabine, a potent nucleoside reverse transcriptase inhibitor. PMID- 11491421 TI - A dose-ranging study to evaluate the antiretroviral activity and safety of amprenavir alone and in combination with abacavir in HIV-infected adults with limited antiretroviral experience. AB - OBJECTIVE: To evaluate the antiretroviral activity and safety of multiple escalating doses of amprenavir administered alone, and in combination with abacavir in HIV-1-infected adults. DESIGN: Sixty-two HIV-1-infected subjects were enrolled in a multicentre, open-label, non-randomized, dose-escalating trial. METHODS: Subjects were assigned to one of six dose groups and received amprenavir 300 mg twice daily, 300 mg three times daily, 900, 1050, or 1,200 mg twice daily for 4 weeks. One dose group received amprenavir 900 mg twice daily in combination with abacavir 300 mg twice daily for 4 weeks. Antiretroviral activity was assessed by measuring changes from baseline in plasma HIV-1 RNA levels and CD4 cell counts. Safety was evaluated by monitoring clinical adverse events and changes in laboratory values. Genotypic and phenotypic analyses were performed using ABI sequencing and the recombinant virus assay, respectively. RESULTS: At week 4, amprenavir monotherapy (900, 1,050, or 1,200 mg twice daily) resulted in marked decreases in plasma HIV-1 RNA levels (1.3-1.6 log10 copies/ml), and substantial increases in CD4 cell counts in the two dose groups who received 1,050 mg twice daily (118 x 10(6) cells/mm3) or 1,200 mg twice daily (114 x 10(6) cells/mm3). Amprenavir/abacavir resulted in median plasma HIV-1 RNA reductions of 1.8 log10 copies/ml, and median CD4 cell count increases of 138 x 10(6) cells/mm3. Amprenavir was reasonably well tolerated with few treatment-limiting adverse events. No known active site mutations associated with amprenavir resistance were selected in any of the dose groups, and no significant phenotypic resistance to amprenavir developed during 4 weeks of therapy. CONCLUSIONS: The antiviral effect of amprenavir monotherapy increased with escalating doses, and all amprenavir doses were reasonably well tolerated over 4 weeks of therapy. Amprenavir/abacavir combination therapy elicited a potent antiviral effect. The three highest doses of amprenavir (900, 1,050 and 1,200 mg twice daily) were selected to design subsequent Phase II and III studies that confirmed the safety profile and efficacy of amprenavir in combination regimens and led to the approval of amprenavir in the USA in 1999. PMID- 11491423 TI - Minimally invasive surgery: brave New World revisited. PMID- 11491422 TI - Adherence over 48 weeks in an antiretroviral clinical trial: variable within patients, affected by toxicities and independently predictive of virological response. AB - OBJECTIVES: To investigate adherence to antiretroviral therapy over 48 weeks, to investigate the association between adherence and treatment-related symptoms and to investigate the impact of adherence on virological response over 48 weeks among established predictors of treatment success. METHODS: One-hundred-and-sixty HIV-1 infected protease inhibitor- and stavudine-naive patients participating in a trial of ritonavir/saquinavir versus ritonavir/saquinavir/ stavudine completed an adherence questionnaire and a symptom checklist at weeks 12, 24, 36 and 48. We calculated odds ratios between experienced symptoms and non-adherence. Regression models were used to determine predictors of HIV-1 RNA below 400 copies/ml at week 48, and of the area about the change from baseline over 48weeks (ACFB) in serum HIV-1 RNA. RESULTS: The percentage of patients reporting missing medication, deviation from time schedule, and dietary prescriptions at separate time-points ranged from 12 to 15%, 32 to 35% and 17 to 22%, respectively. The percentage that changed their level of adherence during 48 weeks ranged from 29% for skipping medication to 48% for deviation from time-schedule. Experienced side-effects were associated with an increased likelihood of non-adherence. Not skipping medication was an independent predictor of both having a serum HIV-1 RNA below 400 copies/ml at week 48 and the ACFB over 48weeks in serum HIV-1 RNA. CONCLUSIONS: Adherence was an independent predictor of virological response over 48 weeks. The level of adherence is variable within patients over time. This suggests the need for continued adherence monitoring in all patients as part of standard medical practice. PMID- 11491424 TI - Outpatient anterior cruciate ligament reconstruction: an analysis of charges and perioperative complications. AB - All outpatient anterior cruciate ligament (ACL) reconstructions using patellar tendon autograft performed at an accredited outpatient surgical center between 1994 and 1998 were prospectively studied. Hospital charges pertaining to the procedures were examined, and perioperative morbidities that might be attributed to an outpatient procedure were evaluated. The study group comprised 284 patients; average patient age at surgery was 28.7 years. Patients were subgrouped into group 1 (isolated ACL reconstructions; n=163), group 2 (ACL reconstructions and meniscal repair; n=48), and group 3 (ACL reconstructions and partial meniscectomy; n=73). Surgicenter facility charges, reoperation rate, complication rate, motion, pain management, hospital emergency room visits, hospital admission, and outpatient surgical facility visits were analyzed. Historical controls from our hospital and our initial outpatient pilot study (May 1994 through November 1995) were used as financial controls. The average surgical center charge for all patients was $3,443. On average, there was a $600 increase for all subgroups from May 1994 through November 1995 compared to December 1995 through August 1998. In the latter time interval, the fixed facility charges were $3,150, $4,075, and $4,275 for groups 1, 2, and 3, respectively. Overall, 19 (7%) patients required a reoperation including 7 (2.5%) patients who required arthroscopic debridement for symptomatic motion deficits. This study expands on our initial published report regarding hospital charges pertaining to an outpatient ACL reconstruction. Extended over another 4 years, we noted slight increases reflective of regional inflationary increases. Compared to our initial inpatient study (1988-1993), significant charge reductions were maintained. This study demonstrated a low complication rate and high patient subjective satisfaction level. PMID- 11491425 TI - Effect of femoral and tibial component position on patellar tracking following total knee arthroplasty: 10-year follow-up of Miller-Galante I knees. AB - Rotational alignment of the femoral and tibial components using computed tomography (CT) was evaluated to establish if errors of alignment have a significant effect on patellofemoral complications. From 1987-1990, 54 knees in 39 patients were replaced with the Miller-Galante I knee system. Of these, 10 patients (13 knees) comprised this study. All patients had osteoarthritis. Mean patient age at surgery was 70.3 years. Mean follow-up was 10.3 years. On CT, rotational position of the femoral component relative to the epicondylar axis and the position of the tibial component relative to the tibial tubercle were evaluated. Patellar displacement and patellar tilt angle also were evaluated when the knee was flexed at 30 degrees. Mean rotation of the femoral component was 6.1 degrees of internal rotation (2.7 degrees-11.2 degrees). Mean rotation of the tibial component was 16.7 degrees of internal rotation (2.4 degrees-27.7 degrees). Mean lateral patellar tilt angle was 2.9 degrees (-6.0 degrees-11.9 degrees), and mean lateral displacement was 2.7 mm (-3.2-8.9 mm). Rotational position of both the femoral and tibial components showed a statistically significant correlation with the patellar tilt angle. This study showed the internally rotated femoral and tibial component were related to the patellar maltracking. This malalignment of the components, as well as nonanatomical patellar groove and metal-backed patellar component, could be one of the causes of the patellofemoral complications with the Miller-Galante I knee. PMID- 11491426 TI - "Custom-fit" versus "off-the-shelf" ACL functional braces. AB - Many sports medicine practitioners believe "custom-fit" functional braces are superior in performance to "off-the-shelf" braces for anterior cruciate ligament (ACL)-deficient knees. However, this is not well substantiated. This study compares a Donjoy custom-fit ACL brace (CE 2000), Donjoy off-the-shelf brace (Goldpoint), and an athletic taping technique to determine their role in our clinical practice. Five patients (3 men and 2 women) with isolated, unilateral, chronic ACL tears with an average age of 27 years (range: 19-35 years) were used to evaluate these three restraint systems. Anterior tibial laxity, quadriceps and hamstrings strength, endurance, standing long jump, brace migration with exercise, and pattern of muscle response to forced anterior tibial displacement were studied. Each patient was tested without a brace and then in each of the three test conditions (custom brace, off-the-shelf brace, and tape), with the order of testing randomized. The Donjoy custom-fit ACL functional brace did not reduce anterior laxity or improve standing long jump, muscle strength, endurance, or muscle response times significantly more than the off-the-shelf ACL brace. Both braces improved anterior stability over knee taping when the knee muscles were contracted under the low forces used in this study. After 1 hour of exercise, brace migration was significantly greater (P=.03) for the CE-2000 custom brace (18.6 mm) than for the Goldpoint off-the-shelf brace (4.5 mm). There appears to be no advantage to the more expensive custom-fit knee brace over the off-the-shelf brace. PMID- 11491427 TI - Observations on patellar height following opening wedge proximal tibial osteotomy. AB - Patellar height and patellar ligament length were assessed pre- and postoperatively in 28 patients who underwent a medial opening wedge proximal tibial osteotomy for varus gonarthrosis. This procedure produced no significant change in patellar ligament length. Pre- and postoperative Insall-Salvati ratios were 0.96+/-0.12 and 0.97+/-0.15, respectively (P=.30). The Insall-Salvati ratio decreased in 29% of patients, and no patient experienced a decline >0.07. The distance between the patella and tibiofemoral joint line ("patellar height") decreased in 100% of patients. The mean Blackburne-Peel ratio declined from 0.75+/-0.13 to 0.53+/-0.15 (P<.001). Sixty-four percent of the postoperative Blackburne-Peel values satisfied the radiographic criterion for patella infera (Blackburne-Peel ratio <0.54). Whereas the loss of patellar height, historically associated with lateral closing wedge proximal tibial osteotomy, is a function of patellar ligament contracture, the decreased distance between the patella and the tibiofemoral joint line following medial opening wedge proximal tibial osteotomy is a function of joint line elevation. The high incidence of patella infera following medial opening wedge proximal tibial osteotomy may have deleterious effects on patellofemoral biomechanics or may complicate subsequent total knee arthroplasty. PMID- 11491428 TI - Neuroanatomic basis for pain in patellar tendinosis ("jumper's knee"): a neuroimmunohistochemical study. AB - Chronic patellar tendinosis or "jumper's knee" is a frequent source of persistent and often recurrent anterior knee pain in active, young people. Seventeen pathologic specimens retrieved during surgery for jumper's knee were evaluated by means of immunohistochemical (S-100) analysis. The patellar tendon-bone junction and the Hoffa fat pad adjacent to the inferior pole of the patella are structures that have a nerve supply that can be a potential source of nociceptive output resulting in the perception of pain at any given moment. In this study, pathologic neural changes also were observed. In eight cases, free nerve endings showed a histologic pattern of "nerve sprouting" in the patellar tendon-bone junction. Vascular innervation was seen in seven cases. S-100 positive fibers were observed within the muscular layer of medium and small arteries. These findings show an increase in vascular innervation. Lastly, neuromatous changes were observed in four cases, demonstrating a clear relationship with pain. These observations provide a neuroanatomic basis for pain in active, young patients with jumper's knee. PMID- 11491429 TI - Angioleiomyoma of the patellar tendon sheath. Case report. PMID- 11491430 TI - Neurilemoma of the tibial nerve causing intermittent claudication. Case report. PMID- 11491431 TI - Failed anterior cruciate ligament surgery: overview of the problem. PMID- 11491432 TI - Revision anterior cruciate ligament reconstruction: technical considerations. PMID- 11491433 TI - Revision anterior cruciate ligament reconstruction: associated patholaxity, tibiofemoral malalignment, rehabilitation, and results. PMID- 11491434 TI - Recent developments in the technical approaches of English-language psychoanalytic schools. AB - This paper summarizes developments in the major approaches to psychoanalytic technique derived from the ego psychology, Kleinian, British independent, self psychology, intersubjectivist, and interpersonal schools over the past fifty years. The author proposes that two major contemporary currents may be differentiated from each other, namely, the psychoanalytic "mainstream"--derived from contemporary Kleinian, contemporary Freudian, and British independent sources, and the "intersubjectivist-interpersonal-self psychology" current. In significant contrast to these two major currents within the English-language psychoanalytic approaches, the French psychoanalytic school has evolved a unique third approach to analytic technique. The author proposes that these three currents constitute the dominant trends regarding technique in contemporary psychoanalytic practice. The paper concludes with a brief outline of the characteristics of each of these technical approaches. PMID- 11491435 TI - On the connection between physical defects and the character type of the "exception". AB - A clinical and theoretical study is presented of the effects of physical defects on character structure, especially on its narcissistic aspects. The basic thesis of this paper is that there are two differentiable responses to awareness of a physical disability: various forms of denial, and a clinging to narcissistic overgratifications as a compensation for negative feelings about the self. The first response, of overusing denial, is universal, and of course leaves its mark on a person's character. However, only the second sort of response, of clinging to overgratifications, leads to the character type of the "exception" (Freud 1916). The distinction between moral ideals, embodied in the ego ideal narrowly defined, and nonmoral ideals, embodied especially in the wishful self-image, is presented as a useful tool in understanding various psychological effects of physical disabilities. PMID- 11491436 TI - Some remarks on adolescence with particular reference to Winnicott and Lacan. AB - Utilizing observations on adolescence--notably those of Winnicott, as well as the work of Lacan and a clinical case--the author advances several propositions concerning the unique relationship between adolescents and time. The consequences of this relationship are then framed as a paternal metaphor. PMID- 11491437 TI - Nontraditional family romance. AB - Family stories lie at the heart of psychoanalytic developmental theory and psychoanalytic clinical technique, but whose family? Increasingly, lesbian and gay families, multiparent families, and single-parent families are relying on modern reproductive technologies to form families. The contemplation of these nontraditional families and the vicissitudes of contemporary reproduction lead to an unknowing of what families are, including the ways in which psychoanalysts configure the family within developmental theory. This article focuses on the stories that families tell in order to account for their formation--stories that include narratives about parental union, parental sexuality, and conception. The author addresses three constructs that inform family stories and that require rethinking in light of the category crises posed by and for the nontraditional family: (1) normative logic, (2) family reverie and the construction of a family romance, and (3) the primal scene. These constructs are examined in tandem with detailed clinical material taken from the psychotherapy of a seven-year-old boy and his two mothers. PMID- 11491438 TI - A consideration of knowledge and authority in the case seminar. AB - This paper examines difficulties in the management of knowledge and authority evidenced in some case seminar settings. The author suggests that the traditional model for the case seminar has not kept pace with evolving ideas about authority and knowledge in the psychoanalytic situation. The tension between our current ways of conceptualizing knowledge and authority in psychoanalysis, and the often unwitting idealization and constriction of knowledge and authority in the case seminar format, are explored. Following a review of the literature on the case seminar, three recommendations for change are discussed: (1) differentiation of the goals of the seminar from those of supervision; (2) reconsideration of the way in which the 'failed case" is discussed; and (3) encouragement of the instructor to present clinical material. PMID- 11491439 TI - Teaching with tape-recorded psychoanalysis. AB - The pedagogic advantages of the study of psychoanalytic process with the help of tape-recorded psychoanalytic sessions is described, with some reference to others' experiences with recording drawn from the literature. Illustration is provided through reference to the reactions of fourth- and fifth-year psychoanalytic candidates to such samplings of one patient's recorded analysis. PMID- 11491440 TI - Surgeons and injury prevention: what you don't know can hurt you! AB - BACKGROUND: The most effective treatment for traumatic injuries is to prevent them from occurring. Currently, few surgeons receive any formal training in injury' control and prevention. This study was designed to test the knowledge of injury prevention principles among practicing surgeons, in order to identify areas in need of intensified educational efforts. STUDY DESIGN: Survey questions designed by members of the American College of Surgeons Committee on Trauma were programmed into a specialized touch-screen computer, which was displayed at four different surgery and trauma meetings, including the ACS Clinical Congress in 1999 and 2000. Participants were questioned about their knowledge of trauma epidemiology, bicycle helmet effectiveness, child safety seat usage, suicide, and domestic violence. RESULTS: Seventy-nine surveys were completed by surgeons, including 33 specializing in trauma care, and by 106 nurses attending trauma courses. Overall, the percentage of correct answers was 50%. There were no significant differences in survey scores between trauma surgeons and general surgeons, although both scored higher than trauma nurses. Areas where knowledge deficits were the most apparent included proper use of child safety seats, the effectiveness of airbags, the prevalence of suicide, and the annual cost of injury in America. CONCLUSIONS: The majority of practicing surgeons and nurses, including those working at trauma centers, are unaware of the basic concepts of injury prevention. Advancements in the field of injury control will require efforts to educate medical professionals and the public. PMID- 11491441 TI - What effect does the duration of an inguinal hernia have on patient symptoms? AB - BACKGROUND: Despite the fact that repair of an inguinal hernia is one of the most common operations performed in general surgery, we have very little information on the natural history of the untreated hernia. The aim of this study was to evaluate the association between hernia symptoms and the duration the patients had their hernias before presentation to a surgical outpatient department for an elective or emergency operation. STUDY DESIGN: Data were gathered prospectively on a consecutive series of 699 patients admitted to two University Departments of Surgery for scheduled operations for an inguinal hernia. RESULTS: More than one third (267) of patients had their hernias for 1 year or longer, up to 65 years, before presentation. The most common symptom on presentation was pain or discomfort at the hernia site, which occurred in 457 (66%) patients. The cumulative probability of pain increased with time to almost 90% at 10 years. The hernia had become irreducible in 48 patients (6.9%). The cumulative probability of irreducibility increased from 6.5% (95% confidence interval 4% to 9%) at 12 months to 30% (95% confidence interval 18% to 42%) at 10 years. Leisure activities were affected in 29% of patients although only 13% of patients had to take time off work because of hernia-related symptoms. Only two patients (0.3%) required resection of infarcted bowel or omentum. CONCLUSIONS: Because many patients with an inguinal hernia are asymptomatic or mildly symptomatic, prospective clinical trials to assess the role of operations for such hernias are required. PMID- 11491443 TI - Longterm results of esophageal myotomy for achalasia. AB - BACKGROUND: Achalasia is a progressive, noncurable, motor disorder of the esophagus. Myotomy of the distal esophagus is the principal method of providing palliation. A major controversy is the necessity for a complementary antireflux procedure. STUDY DESIGN: Forty-two patients were studied by clinical history manometrically, roentgenographically, and endoscopically. Transabdominal Heller myotomy is the preferred approach. Nine patients had Nissen fundoplication and parietal cell vagotomy (group 1), and 16 had posterior gastropexy and parietal cell vagotomy (group II). Initially 16 of 17 patients underwent transthoracic Heller myotomy without fundoplication (group III). Twenty-five patients were followed a mean of 10 years (range 5 to 26 years). RESULTS: One postoperative death was from adult respiratory distress. Results in group I were excellent in five, good in three, and fair in one. The patient with a fair result developed a diverticulum at the myotomy site and significant reflux at 9 years. Results in group II patients were excellent in 2, good in 11, there was 1 operative death, and no followup in 1. Of the 17 patients in group III, 3 had resection of an esophageal diverticulum, and 3 had closure of esophageal perforation caused by pneumatic dilatation. Results in the 13 patients followed were excellent in 6, good in 5, and poor in 2. CONCLUSIONS: There is no statistical difference in results by chi-square analysis between transthoracic Heller myotomy without fundoplication and transabdominal Heller myotomy with parietal cell vagotomy and Nissen fundoplication or posterior gastropexy. PMID- 11491442 TI - Soft tissue sarcomas of the groin: diagnosis, management, and prognosis. AB - BACKGROUND: Sott tissue sarcomas (STS) of the groin may present a difficult problem because or misdiagnosis as groin hernia and proximity to major neurovascular structures. We evaluated our management and survival in a large cohort of patients. STUDY DESIGN: Patients treated between July 1, 1982 and July 1, 1998 with primary or recurrent STS of the groin were included. Groin sarcomas were defined as those tumors within 5 cm of the inguinal crease. Patient, tumor, clinical, and survival data were analyzed using a log rank test and Cox regression. RESULTS: We treated and followed 88 patients with STS of the groin. The median age was 52 years (range 16 to 86 years) and 55 patients (63%) were male. Disease-specific survival was 72% at 5 years. Tumors tended to be larger than 5 cm (52%), deep (72%), and high-grade (60%). Unfavorable prognostic factors for disease-specific survival were high grade (p < 0.001), neurovascular invasion (p < 0.001), positive margin (p < 0.01), deep depth (p < 0.01), and selection for adjuvant therapy (p < 0.005). Multivariate analysis indicated age greater than 50 years (p < 0.05), high grade (p < 0.001), neurovascular invasion (p < 0.001), and positive microscopic margins (p < 0.001). Fourteen patients (16%) were diagnosed with STS at hernia operation then went on to a definitive operation with no impact on survival. Seventeen patients (19%) had involvement of a major vessel or nerve, and 5 of these ultimately required amputations, 3 for local recurrence. CONCLUSIONS: High grade, neurovascular invasion, and positive microscopic margins are associated with poor outcomes. The biology of these tumors is similar to other extremity STS, and similar principles of management apply. Even with neurovascular involvement, most patients with primary groin STS do not require amputation. PMID- 11491444 TI - Serial computed tomography is rarely necessary in patients with acute pancreatitis: a prospective study in 102 patients. AB - BACKGROUND: CT has proved to be helpful in patients with acute pancreatitis for differentiating between mild and severe forms. Followup of acute pancreatitis with CT has been advocated but rarely studied. The aim of this study was to determine if late CT performed at day 7 might be helpful in establishing the prognosis or the type of complications, and to select a subgroup of patients in whom CT could be beneficial. STUDY DESIGN: Contrast-enhanced CT was performed at the admission day and 7 days after admission in 102 patients admitted for acute pancreatitis. The extent of pancreatic inflammation was classified according to Balthazar grade, and intrapancreatic necrosis on these examinations was prospectively assessed and compared with clinical and biologic data and with patient outcomes. RESULTS: Among 102 patients, complications developed in 24 (23%). Complications developed in only 8% of patients with Ranson score <2, making routine early CT unnecessary. For the patients with Ranson score <2 and Balthazar grades A and B at day 1 CT, late CT seemed to be useless. Complication was suspected by clinical and biologic tests before day 7 in 22 of 24 complicated patients (92%), suggesting that CT could be proposed only in cases of clinical or biologic deterioration. Late CT was correlated with a complicated course in patients with Balthazar grades D and E or intrapancreatic necrosis >50%. Late CT was predictive of complications in cases of intrapancreatic necrosis enlarging since the first examination. CONCLUSIONS: Our study showed that in acute pancreatitis: 1) there is little justification for systematic early CT, especially in patients with Ranson score <2, and 2) late CT does not need to be performed routinely, but only in cases of clinical or biologic worsening. PMID- 11491445 TI - Simultaneous detection of colorectal carcinoma liver and lung metastases does not warrant resection. AB - BACKGROUND: Recent evidence suggests that metastasectomy is efficacious for selected patients with hepatic and pulmonary metastases from a colorectal primary. The aim of this study was to identify a subgroup of patients who best benefit from hepatic and pulmonary metastasectomy among those with colorectal carcinoma metastases. STUDY DESIGN: We analyzed retrospectively a total of 136 patients who underwent resection of hepatic or pulmonary metastases of colorectal origin at Niigata University Medical Hospital between 1982 and 2000. The median follow-up period was 94 months. Eighty-four patients underwent hepatectomy alone, 25 underwent pulmonary resection alone, and 27 underwent both hepatic and pulmonary resection. The 27 patients undergoing hepatic and pulmonary resection were divided into two groups: 17 patients with sequentially detected hepatic and pulmonary metastases and 10 patients with simultaneously detected metastases. Survival time was determined from the date of initial metastasectomy. Differences in cumulative survival were evaluated using the log-rank test. Sixteen factors were assessed for their influence on the survival of the 27 patients undergoing resection of hepatic and pulmonary metastases; univariate and multivariate analyses were used in this evaluation. RESULTS: Patient survival after hepatic and pulmonary resection was comparable with that after hepatectomy alone (p = 0.536) and that after pulmonary resection alone (p = 0.294). Among the 27 patients undergoing hepatic and pulmonary resection, the outcomes after resection were significantly better in patients with sequentially detected metastases (cumulative 5-year survival of 44%) than in those with simultaneously detected ones (cumulative 5-year survival of 0%) (p < 0.001). On multivariate analysis sequential detection of hepatic and pulmonary metastases was the strongest independent favorable prognostic factor (p <0.001). CONCLUSIONS: Patients with sequentially detected hepatic and pulmonary metastases from a colorectal primary are good candidates for aggressive metastasectomy. Simultaneous detection of these metastases does not warrant resection. PMID- 11491446 TI - Treatment of hemorrhoids with circular stapler, a new alternative to conventional methods: a prospective study of 140 patients. AB - BACKGROUND: Surgical hemorrhoidectomy has a reputation for being a painful procedure. The aim of this study was to determine the efficacy and safety of a new procedure for surgical treatment of hemorrhoid disease. STUDY DESIGN: From April 1998 to August 1998, 140 patients (83 men and 57 women) with an average age of 43.8 years (range 19 to 83 years) underwent hemorrhoidectomy using a circular stapler. Operative times, pen- and postoperative complications, mean hospital stay, assessment of the postoperative pain, period of incapacity for work, and functional results were collected. All patients were evaluated at 2 weeks, 2 months, and 18 months after operation. RESULTS: The average length of the operation was 18 minutes (range 8 to 60 minutes). There were no perioperative complications. The postoperative complication rate was 6.4% (n = 9). Mean hospital stay was 36 hours (range 8 to 72 hours). Paracetamol was the only analgesic used. Eighty-three patients (59.3%) required analgesic for less than 2 days, 45 patients (32.1%) between 2 and 7 days, and 12 patients (8.6%) more than 7 days. No patients had anal wound care. One hundred four patients had professions. The period of incapacity for work was less than 3 days for 22 patients (21.1%), between 3 and 7 days for 13 patients (12.5%), between 7 and 14 days for 62 patients (59.6%), and more than 14 days for 7 patients (6.8%). At 18 months, 95.7% of patients were fully satisfied with the results, 3.6% were somewhat satisfied (n = 4), and 0.7% were unsatisfied. CONCLUSIONS: Treatment of hemorrhoids with a circular stapler appears to be safe, effective, and rapid, causing few postoperative complications and minimal postoperative pain. At 18 months, 95.7% of the patients were fully satisfied with the results. PMID- 11491447 TI - Current indication of a modified Sugiura procedure in the management of variceal bleeding. AB - BACKGROUND: The role of gastroesophageal devascularization (Sugiura-rype procedures) for the treatment of variceal bleeding remains controversial. Although Japanese series reported favorable longterm results, the technique has nor been widely accepted in the Western Hemisphere because of a high postoperative morbidity and mortality. The reasons for the different outcomes are unclear. In a multidisciplinary team approach we developed a therapeutic algorithm for patients with recurrent variceal bleeding. STUDY DESIGN: The Sugiura procedure was offered only to patients with well-preserved liver function (Child A or Child B cirrhosis without chronic ascites) who were not candidates for distal splenorenal shunt, transhepatic porto-systemic shunt, or liver transplantation. RESULTS: Fifteen patients with recurrent variceal bleeding underwent a modified Sugiura procedure between September 1994 and September 1997. All but one patient (operative mortality 7%) are alive after a median followup of 4 years. Recurrent variceal bleeding developed in one patient; esophageal strictures, which were successfully treated by endoscopic dilatation, developed in three patients; and one patient experienced mild encephalopathy. Major complications were noted only in patients with impaired liver function (Child B cirrhosis) or when the modified Sugiura was performed in an emergency setting. The presence of cirrhosis or the cause of portal hypertension had no significant impact on the complication rate. CONCLUSIONS: This series was performed during the last decade when all modern therapeutic options for variceal bleeding were available. Our results indicate that the modified Sugiura procedure is an effective rescue therapy in patients who are not candidates for selective shunts, transhepatic porto-systemic shunt, or transplantation. Emergency settings and decreased liver function are associated with an increased morbidity. PMID- 11491448 TI - The quest for painless surgical treatment of hemorrhoids continues. PMID- 11491449 TI - What's new in urology. PMID- 11491450 TI - What's new in endocrine surgery. PMID- 11491451 TI - Review of sentinel lymph node credentialing: how many cases are enough? PMID- 11491452 TI - A historic perspective of liver surgery for tumors at the end of the millennium. PMID- 11491453 TI - Small bowel intussusception from malignant posttransplantation non-Hodgkin's lymphoma. PMID- 11491454 TI - Frantz's tumor. PMID- 11491455 TI - Laparoscopic continuous hyperthermic peritoneal perfusion. PMID- 11491456 TI - Human error. PMID- 11491457 TI - Assessing competency. PMID- 11491458 TI - Antibacterial and anticholinesterase activities of aplysamine-4, a bromotyrosine derived metabolite of a Red Sea marine sponge. AB - Aplysamine-4, a metabolite of likely bromotyrosine biogenesis, was isolated from an unidentified verongid sponge from the Red Sea. The compound was identified by heteronuclear magnetic resonance experiments, and by electrospray ionization tandem mass spectrometry. The compound exhibited moderate inhibitory activity on several Gram positive and Gram negative bacteria, and was also found to be a non competitive reversible inhibitor of acetylcholinesterase. At pH 7.4, a Ki value of 16 and 2 microM was determined with electric eel and insect recombinant acetylcholinesterase, respectively. A deprotonated form of aplysamine-4 was obtained by alkaline treatment of the natural compound and it was shown to be less active than the protonated form. PMID- 11491459 TI - Sodium current in NG108-15 cell inhibited by scorpion toxin BmKAS-1 and restored by its specific monoclonal antibodies. AB - Effect on Na+ current of BmKAS-1, a novel polypeptide purified from the venom of Chinese scorpion Buthus martensi Karsch (BmK), has been investigated in differentiated NG108-15 cells by using patch-clamp whole-cell recording. Neutralizing effect of four monoclonal antibodies of BmKAS-1 (mAb #2, #3, #4, and #5) has also been observed. The results showed that Na+ current was irreversibly inhibited by BmKAS-1 and the inhibitory effect was abolished by mAb #2 and #5, but not by mAb #3 and #4. PMID- 11491460 TI - Isolation, purification and N-terminal sequencing of a bioactive peptide that alters action potentials from the venom of Buthus martensii Karsch. AB - A bioactive peptide that extensively prolongs action potentials (APs) in frog nerve has been isolated and purified from the venom of the scorpion Buthus martensii Karsch (BMK). The peptide, designated as BMK 18(2), was purified using gel filtration, ion exchange, FPLC, and HPLC chromatography. APs recorded in the presence of nanomolar concentrations of the peptide were extensively prolonged with some attenuation in their heights. The N-terminal sequence of BMK 18(2) was found to be: VRDAYIAEDYD-VYH-ARDA. Sequence similarity comparisons to other alpha scorpion toxins suggest that the two blanks in the sequences are cysteines. The molecular weight (M.W.) of BMK 18(2) was determined by LC/MS/MS to be 7185 Da. Since the peptide prolongs APs when both K+ and Ca++ channels were blocked and shows sequence similarity to other alpha-neurotoxins, it appears likely that BMK 18(2) acts to alter Na channel inactivation to produce its effect. PMID- 11491461 TI - Isolation and characterization of nerve growth factor (NGF) excreted from cultured eukaryotic cells. AB - Since the discovery in 1954, NGF has been isolated from snake venoms and various tissues and organs of different animals. Recently, Lipps (2000b) reported the isolation of NGF from the human body fluids, saliva, serum, and urine. This investigation reports the isolation of NGF excreted by various types of cells from diverse origin, in serum free medium, proving that its presence is not restricted to neural cells. The established cell lines used were Chang's liver and neuroblastoma of human origin, Vero monkey origin, PC12 rat, and SP/2 mouse origin. The fully grown monolayers of the cells were maintained in serum free medium for 48 hours to excrete NGF in the medium and the cell free medium was concentrated. NGF from cell free concentrated medium for each cell line was isolated by high pressure liquid chromatography (HPLC) and was identified as described by Lipps (1998). The HPLC profiles for cell free concentrate from different types were observed to be similar. The NGF fraction was eluted last in the neutral pH (Fig. 1). The identified fraction for NGF from each cell line was further purified, which resolved into a single peak. The purified NGF was used to study the biological and immunological properties. The biological activities of NGFs from cell free medium were minuscule in comparison to the cobra venom derived NGF. The molecular weights of NGFs from cell free medium for all cell lines were identical, 36.0 kDa. Anti-human NGF reacted strongly immunologically with NGFs from human origin cells and poorly with rat and mouse. PC12 NGF antibody reacted immunologically only with PC12 NGF. PMID- 11491462 TI - Characterization of a thrombin-like serine protease, Kangshuanmei, isolated from the venom of a Chinese snake, Agkistrodon halys brevicaudus stejneger. AB - An enzyme, referred to as Kangshuanmei, was isolated from the venom of the Chinese snake Agkistrodon halys brevicaudus stejneger by gel filtration chromatography followed by affinity chromatography. Kangshuanmei is composed of a single polypeptide chain with a molecular weight of approximately 34,000, estimated by SDS-PAGE. The enzyme hydrolyzed both benzoyl-arginine ethyl ester and H-D-Phe-Pip-Arg-p-nitroanilide, specific substrates for thrombin. The protease activity of Kangshuanmei was inhibited by 4-(2-aminoethyl) benzensulfonyl fluoride, but was not affected by EDTA. The enzyme acted on human fibrinogen to form a fibrin clot and released three fragments. These fragments were shown to be fibrinopeptide A, fibrinopeptide B, and the Bbeta1-42 peptide of fibrinogen, respectively. These results indicate that Kangshuanmei is a thrombin like serine protease with coagulant activity. However, the enzyme did not induce activation of blood coagulation factor XIII, unlike thrombin. Moreover, antithrombin-III, the specific thrombin inhibitor in plasma, had no inhibitory effect on the thrombin-like amidolytic activity of Kangshuanmei. The N-terminal amino acid sequence of the enzyme up to 50 residues was determined by a peptide sequencer. The N-terminal sequence of Kangshuanmei was highly homologous to most thrombin-like serine proteases from the venom of the snakes of the crotalidae family. PMID- 11491463 TI - Thrombin-like enzyme, flavovilase, with kinin-releasing activity from Trimeresurus flavoviridis (habu) venom. AB - A thrombin-like enzyme, flavovilase, with kinin-releasing activity was isolated, purified, and characterized from the venom of Trimeresurus flavoviridis (habu) using Sephadex G-100, DEAE-Cellulose, and CM-Cellulose column chromatographies. The final preparation was homogeneous as demonstrated by a single band on polyacrylamide gel electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and isoelectric focusing electrophoresis. The enzyme possesses a molecular weight of 26,500, an isoelectric point of 5.0, and consists of 247 total amino acid residues. Specific electrolytic activities of this enzyme on N tosyl-L-arginine methyl ester (TAME) and N-benzoyl-L-arginine ethyl ester (BAEE) were determined to be 50.9 and 17.4 micromol/min/mg, respectively. The enzyme was inhibited by p-APMSF (p-amidinophenylmethanesulfonyl fluoride hydrochloride), beta-mercaptoethanol, and N-bromosuccinimide. Additionally, the enzyme was found stable to heat treatment. It was also observed that the enzyme cleaved a kininogen analog with the release of bradykinin. PMID- 11491464 TI - Effectiveness of Thai cobra (Naja kaouthia) antivenom against sea snake (Lapemis hardwickii) venom: verification by affinity purified F(AB')2 fragments. AB - Commercial Thai cobra (Naja kaouthia) antivenom was found to be effective in neutralizing sea snake (Lapemis hardwickii) venom. Neurotoxin specific F(ab')2 fragments obtained from the antivenom by chromatography using Thai cobra neurotoxin or sea snake venom affinity columns were able to neutralize both venoms. PMID- 11491466 TI - These changing times. PMID- 11491465 TI - Binding to and hemolysis of human erythrocytes by pyrularia thionin and Naja naja kaouthia cardiotoxin: inhibition by prothrombin. AB - Pyrularia thionin and snake venom cardiotoxin are strongly basic proteins which bind to and induce hemolysis of erythrocytes, cause depolarization of muscle cells, and influence the order and properties of phospholipids in cellular membranes. Earlier studies showed a competition between the thionin and cardiotoxin for a common binding site on erythrocytes, and the present study extends these studies to show a similar competition between prothrombin and both basic proteins. The competition between the thionin and prothrombin for binding sites on erythrocytes was shown by direct binding experiments using radiolabeled thionin. Whereas binding of thionin or cardiotoxin induces hemolysis as a consequence of membrane perturbation, prothrombin does not induce hemolysis. Although it binds to the same site, there is no penetration into and perturbation of the membrane. The competition between prothrombin and pyrularia thionin is not influenced by added Ca++. This indicates that membrane PS interacts in a specific and Ca++-independent manner with at least one site on prothrombin, as proposed earlier (Tendian and Lentz, 1990). The competition between thionin and prothrombin was also demonstrated for the release of radiolabeled chromate from loaded P388 cells. The competition observed with the P388 cells shows that the competition is not a unique phenomenon with erythrocytes, but occurs with other cell membranes. PMID- 11491467 TI - Gathering information 2: methods of assessment. PMID- 11491468 TI - NVQ in oral health care for dental nurses. PMID- 11491469 TI - NHS plan--proposal for new approach. PMID- 11491470 TI - Evolution of occlusion, past and present time. PMID- 11491471 TI - Milk, flavoured milk products and caries. AB - The consumption of flavoured milk increased by 50% between 1992 and 1999 and dental health educators need to know if these and other sugar and fruit juice sweetened milk products, such as fruit yoghurts, are acceptable as snack items. Available evidence suggests that their cariogenicity is negligible to low and consumed in moderation they are a preferable alternative to similarly sweetened soft drinks. PMID- 11491472 TI - Chronic idiopathic orofacial pain: I: What is the evidence base? AB - Chronic idiopathic orofacial pain is currently a problem for the specialists, but is this type of disorder peculiar to the face, and is there a role for the general dental practitioner? This article reviews the current understanding and management of medically unexplained syndromes and discusses their application in facial pain. A second article will merge this contemporary theory and clinical evidence into a management strategy, and stress the importance of early intervention in the primary care setting. PMID- 11491473 TI - Don't forget your toothbrush! AB - Patients with rectally inserted foreign bodies can present to accident and emergency departments or general medical practitioners. Rarely dentally related objects are inserted because of their ready availability in the domestic environment. There are many reasons given for their presence in the rectum, most commonly accidental insertion, assault, and psychosexual motives. This case is the first reported incident of a patient using a toothbrush to relieve his pruritus ani and subsequently losing it up into the rectum. PMID- 11491474 TI - VDP survey. PMID- 11491475 TI - Implant treatment. PMID- 11491476 TI - Consent: the patients' view--a summary of findings from a study of patients' perceptions of their consent to dental care. AB - AIM: A study was carried out which aimed to investigate peoples' perceptions of how their consent was given for dental treatment. METHOD: A structured questionnaire was completed by 50 patients receiving treatment in the British NHS and 12 of them completed an in-depth recorded interview. The data was analysed using a combination of quantitative and qualitative methods. RESULTS: The findings showed that 80% of patients had not been given any written treatment plan (FP17DC). 79% of patients mistakenly thought that the form signed requesting NHS treatment (FP17) was a consent form. When options were offered patients were more likely to consider that they were involved in the consenting process. However consent is still often implied rather than explicit. There was a range of experience reported by patients from those who felt that the dentist made the treatment decisions to those who felt that decisions had been made collaboratively. Although some patients were happy with the way their consent was obtained examples were also given of lack of information, confusion and even of deceit. CONCLUSIONS: There is an urgent need to clarify the status of NHS documentation regarding consent and a general need for awareness to be raised in the dental profession about the importance of obtaining consent which is freely given based on appropriate information which has been adequately understood. PMID- 11491477 TI - It's only teething...a report of the myths and modern approaches to teething. AB - Paediatric dentistry is not my usual field of work. I am now based almost entirely in restorative dentistry and it is five years since I worked in the dental department of a children's hospital. An essay on teething would appear to be an unusual choice of topic. With the current professional climate of 'general professional education' and 'lifelong learning' I can easily justify my time and effort studying a subject somewhat removed from my regular work. However, to be completely honest, I have reached that age when many of my friends, relatives and colleagues are enjoying the sleepless nights that accompany expanding families. Add to this the fact that I have recently married into a family of midwives, health visitors, nurses and new mothers. I was not sure that I was giving the best, most up to date advice when asked about teething. So some reading around was required. If only it were that simple. I now feel equipped to give a little more help than simply saying, "It's only teething..." PMID- 11491478 TI - Modern methods for assessing the cariogenic and erosive potential of foods. AB - Assessment of the acidogenic and cariogenic erosive potential of foods requires the use of reproducible and standardised methods. The methodology laid down at the San Antonio conference in 1985 remain today the basis for research on and the testing of foods. The 1999 workshop has updated the appropriate methodology and introduced guidelines on the testing of erosive potential of foods. It is to be hoped that researchers wishing to evaluate foods and their effect on the teeth will continue to use these methods. PMID- 11491480 TI - 52 and counting. PMID- 11491479 TI - The Commonwealth Oral Health Statement. PMID- 11491481 TI - Clinical diagnostic and intervention studies of children with semantic-pragmatic language disorder. AB - The diagnosis of semantic-pragmatic language disorder (SPLD) has been the subject of a number of research studies over the last two decades. Classification and diagnostic debates, while illuminating, have done little to develop tools to improve services to these children. In this paper, two children whose communication difficulties are suggestive of an SPLD diagnosis but who have differing profiles are studied. Using existing models of psycholinguistics and pragmatics to guide assessment and intervention, the diversity of language and social communicative behaviours that are covered by the label SPLD are exemplified. Consideration is given to whether the term SPLD is appropriate for both children or whether Bishop's revision of the diagnosis to 'pragmatic language impairment' might be more useful. Methods of intervention and evaluation for semantic and pragmatic deficits in these two cases are described. It is argued that existing tools can enable accurate explanation and modelling of the communication of children with SPLD and that there is a role for intervention studies in helping to refine those tools, to improve therapies and to understand the nature of the condition more fully. PMID- 11491482 TI - Comprehension of inferential meaning in language-impaired and language normal children. AB - Three groups of language-normal (LN) 6, 8 and 16/17 year olds, and a group of language-impaired (LI) children were given a task answering questions about pictures that involved inferential meaning. A developmental progression in the types of responses given is seen, with the LI children performing like the youngest LN children. A similar progression is seen in the ability to justify the answers given to inferential questions with the young adult group giving the fewest justifications that were problematical in some way. Larger numbers of problematical justifications in the LI group can be related to some extent to non pragmatic aspects of their impairment, but overall this group also gave more pragmatically irrelevant responses when asked to justify answers given. It was not possible to identify any major differences between subgroups of children within the LI group, identified as pragmatically impaired (PI) and non pragmatically impaired (NPI), in terms of either answers given to inferential questions or in terms of problematical justifications for these answers. Possible reasons for these findings are discussed. PMID- 11491483 TI - Consensus and variation between speech and language therapists in the assessment and selection of preschool children for intervention: a body of knowledge or idiosyncratic decisions? AB - The aim was to investigate consensus between speech and language therapists (SLT) during the initial assessment of preschool children. A multimethod design was used. The qualitative component provided SLT's own views of the process as the starting point for the study. Data were generated from semistructured interviews and focus group discussions. The interviews investigated factors considered during an initial assessment of a preschool child. The focus groups discussed the process and content of a videotaped initial assessment carried out by the author. Quantitative data was generated from six case analyses and from a card sorting exercise. In the former, SLT were asked to record important features of newly referred cases. In the latter, SLT sorted statements about preschool children according to a simple concern/not concerned decision. Data collection tasks and analysis took place successively to build a composite and triangulated picture. The main subjects were 11 SLT, experienced in childrens' work with a further six acting as a broader reference group. The results show an interacting pattern of consensus and variation. SLT agreed about the relative priority of children, categories that informed the assessment; they made similar interpretations of incoming information. There was variation in their preferred assessment methods, in the thresholds of concern and in the interventions offered. Sources of variation that were identified included individual confidence in and access to assessments, personal levels of concern and in the approach to the SLT's role, as well as the influence of workplace policies. The study concluded that, consensus and variation both exist within the context of SLT's initial assessment of preschool children, but that there are discernible patterns to the variation. While such variation may be inevitable, it is suggested that any variation should be made explicit and used to challenge and develop the shared knowledge base of the speech and language therapy profession. PMID- 11491484 TI - Use of cognitive state predicates by language-impaired children. AB - Two studies of the use of cognitive state predicates by children with specific language impairment (SLI) were conducted. Study I analysed longitudinal language samples collected from 26 children with SLI and 25 children with normal language (NL) development, aged 4;4 and 2;11, respectively, at Time I. Study II analysed samples from SLI children with more severe delays at an earlier language stage. There were 10 SLI children and 10 NL children, aged 4;11 and 2;8, respectively, matched by MLU. All cognitive state predicates were identified using both broad and narrow definitions. In Study 1, the SLI children used cognitive state predicates less frequently than their mental age peers, and with no greater frequency or variety than their younger, language peers. In Study II, children with SLI used more predicates referring to communication events, but there were no further group differences. These findings are discussed as they relate to two current psycholinguistic issues: the possible dissociation of grammar and the lexicon, and the role of language in the development of children's theory of mind. PMID- 11491485 TI - Interaction between children with cerebral palsy and their mothers: the effects of speech intelligibility. AB - Children with cerebral palsy and severe speech disorders have been described as passive communicators, whose conversation partners control and dominate interaction. This paper studies the effects of speech intelligibility on interaction between mothers and their children with cerebral palsy to examine if similar patterns of child passivity and adult domination are also observed in dyads in which children are physically dependent but naturally intelligible to their parents. Two groups, each of 20 children who had four-limb cerebral palsy and who differed in their speech intelligibility, participated in the study. Children in Group 1 (whose interaction has been previously reported) were unintelligible to their parents out of context. Children in Group 2 had speech that was understood without situational cues. Children and their mothers were videotaped playing with a set of toys that had been shown to elicit from non disabled children a full range of the conversational moves and speech acts targeted in the study. Videotaped interaction was transcribed and coded at three levels to show conversation structure, the pragmatic functions expressed and the method of communication used. Percentage distribution of coding categories was compared across groups using coda. Results showed that verbally intelligible children initiated more conversations and used their communication for a wider range of functions than did non-speaking children, for whom more restricted patterns of conversation were noted. Unexpectedly, few differences were observed between the two groups of mothers, who initiated and closed most exchanges. Implications for intervention are discussed. PMID- 11491486 TI - Non-word repetition and grammatical morphology: normative data for children in their final year of primary school. AB - It is likely that tests of phonological short-term memory (non-word repetition) and grammatical morphology (past tense, third-person singular) are clinical markers for specific language impairment (SLI). The study provides normative data on three specific tests covering the aforementioned areas for 100 children attending the final year of primary schooling (10-11 year olds). Data from typically developing age controls can be used to provide a benchmark against which to compare the ability of language-impaired children. This is particularly important given that the transition from primary to secondary schooling is a key stage in a child's education. PMID- 11491487 TI - Re: Stuttering therapy results in pseudofluency. PMID- 11491488 TI - Re: Frequency altered feedback as an alternative to 'prolonged speech' techniques for the control of stuttered speech. PMID- 11491489 TI - Re: A commentary on pseudofluency. PMID- 11491490 TI - Erythema nodosum: a clinical approach. PMID- 11491491 TI - Stress of chronic food restriction attenuates the development of adjuvant arthritis in male Long Evans rats. AB - OBJECTIVE: To study the effect of the stress of chronic food restriction on the development of adjuvant arthritis in Long Evans male rats. METHODS: Four groups of animals were compared: non-treated control (C) and arthritic (AA) rats, both with free access to food and water and two analogous groups with a 40% food restriction, i.e. non-treated (FR) and arthritic (AA-FR) animals. All rats were killed 22 days following the injection of complete Freund's adjuvant. The parameters measured were: serum levels of albumin (ALB), nitrate, glucose, insulin, corticosterone (CORT), prolactin (PRL) and PRL mRNA in the adenopituitaries. In addition the activity of gamma-glutamyl transpeptidase (GGTP) was measured in the spleen. The pain threshold was determined by the tailflick method. The body weight of the animals was recorded on day 0, 3, 7, 11, 15 and 18 of the disease. RESULTS: Arthritis caused swelling of the hind paw (2.37 +/- 0.15 ml vs 1.1 +/- 0.05ml in controls, p < 0.01) which was prevented in the AA-FR group (1.44 +/- 0.13 ml, not significant against controls). Arthritis increased serum NO and reduced ALB levels; both changes were significantly restored in the FR-AA group. Food restriction did not alter the activation of GGTP, or the decrease of PRL mRNA observed in the AA group. Serum CORT was elevated in rats with food restriction (15.49 +/- 2.1 vs. 3.7 +/- 0.87 microg/dL) and remained enhanced to the same extent in AA and FR-AA groups. The tailflick latency prolonged in the AA group was reduced by food restriction. CONCLUSION: These results show that 40% food restriction associated with elevated CORT levels mitigated the inflammatory parameters activated during AA. PMID- 11491492 TI - Intra-articular injection of hyaluronate (SI-6601D) improves joint pain and synovial fluid prostaglandin E2 levels in rheumatoid arthritis: a multicenter clinical trial. AB - OBJECTIVE: The relationship between clinicalfeatures and biochemical parameters of synovialfluid after serial intra-articular injections of sodium hyaluronate (SI-6601D) was investigated. METHODS: SI-6601D (sodium hyaluronate with an average molecular weight of 8.4 x 10(5); 25mg/2.5ml/syringe) was injected intra articularly into the knees of 25 patients with rheumatoid arthritis (RA) every week for 5 consecutive weeks. Clinical and biochemical parameters were monitored before and after injection. Clinicalfindings included pain, as a summation of 3 categories (pain at rest, pain in motion and pain in passive motion, each assessed on a 4-step rating scale), and inflammation, also as a summation of 3 categories (swelling, patellar ballotement and local warmth, each assessed on a 4 step rating scale). Pain on walking of patient was qualitatively assessed by visual analogue scale (VAS). The aspirated volume of synovialfluid (SFV) was recorded and levels of prostaglandin (PG) E2, transforming growth factor beta-1, tumor necrosis factor alpha, interleukin I receptor antagonist, chondroitin 4 sulfate (C4S) and chondroitin 6-sulfate were measured. RESULTS: Significant improvement in pain symptoms (p < 0.0001), inflammation (p < 0.0001), VAS pain (p < 0.001) and SFV (p < 0.05) were observed after the 5 injections. Levels of PGE2 (p < 0.05) and C4S (p < 0.05) in the synovialfluid were significantly decreased. DISCUSSION: SI-6601D improved local clinical symptoms in RA patients by suppressing PGE2 and, therefore, may be a useful treatment for local inflammation in RA. PMID- 11491493 TI - Immunization with TCR Vbeta10 peptide reduces the frequency of type-II collagen specific Th1 type T cells in BUB/BnJ (H-2q) mice. AB - OBJECTIVE: Collagen induced arthritis (CIA) in mice is mediated by synergistic T cell and humoral immune responses specific for type II collagen (CII). We have previously shown that in arthritic joints of BUB mice (TCR Vbetaa, H-2q) the TCR repertoire is enrichedfor Vbeta10 expressing T cells, and that immunization with a Vbeta10 peptide (Vbeta10p) prevents the phenotypic expression of disease. The objective of the present study was to understand how immunization with a synthetic TCR Vbeta peptide affected the development of the pathogenic CII specific immune response in BUB mice. METHODS: Arthritic and protected animals were tested for Vbeta10p- and CII-specific cytokine production by a highly specific and sensitive ELISA spot assay, andfor CII-specific antibody production by standard ELISA. In adoptive transfer experiments, Vbeta10p-specific LN cells (INF-gamma producing) were injected into naive mice prior to immunization with type-II collagen/CFA. RESULTS: Immune cells from arthritic animals produced IFN gamma and IL-2, without IL-4 and IL-5 in response to CII and an immunodominant epitope, A2, derivedfrom CII. Serum from these mice contained anti-CII antibodies of both IgGI and IgG2a subtypes. Our results show for thefirst time that immunization with Vbeta10p resulted in Vbeta10p-specific IFN-gamma and IL-2 production that was restricted to the CD4+ T cell subset. Emergence of this Vbeta10p-specific immune response was associated with a dramatic decrease in the frequency of CII and A2-specific, cytokine producing T cells in arthritis protected mice. Protective immunity was cell mediated and could be adoptively transferred. In contrast, the protective immunization had only a marginal effect on the anti-CII antibody response indicating that the CII specific humoral immune response was not significantly affected. CONCLUSION: Immunization with TCR Vbeta10p leads to expansion of a population of Vbeta10p- specific CD4+ Tcells. This anti-TCR Vbeta10p specific type 1 cytokine producing immune response was protective in adoptive transfer studies and appears to inhibit the expansion of the pathogenic anti-CII cellular immunity. Additionally, the anti-TCR Vbeta10p specific cellular immune response was mediated by CD4+ T cells and these T cells did not produce IL-4 or IL-5. Thus, our results suggest that protection against CIA in mice immunized with synthetic TCR Vbeta10p was achieved by a specific down regulation of the CII-specific Thl type cellular immune response and not via immune deviation. PMID- 11491494 TI - Risk factors associated with mortality in systemic lupus erythematosus. A case control study in a tertiary care center in Mexico City. AB - OBJECTIVE: To identify the mortality risk factors in a group of Mexican patients with SLE. METHODS: A case-control autopsy study in a tertiary care center in Mexico, City. Patients with SLE who died during 1958 to 1994 with an autopsy study were selected as cases, and alive patients matched by age (+/- 3 years), decade of SLE onset, and disease duration (+/- 5 years) were defined as controls. Clinical charts were reviewed looking at clinical variables. SLE disease activity was evaluated with the MexSledai index, and SLE disease severity with the Severity Index. Variables were classified as present at any moment during the follow-up and 3 months before death in cases or cut-off date in controls. STATISTICAL ANALYSIS: matched univariate and multivariate analysis by multiple logistic regression were performed, and the results were presented as odds ratio and 95% confidence intervals (OR, 95%CI). RESULTS: 76 matched pairs of patients were studied. Age, gender, and years offormal education were similar in the cases and controls. Variables associated with mortality three months before death were: lung involvement OR= 15.6, 95%CI (4.8-50.3), p<0.001; severe thrombocytopenia 9.6 (2.9-31.7), p<0.001; heart involvement 5.8 (2.6-13.0), p<0.001; and the severity index (cases 8.8 mu, 2.4 sigma vs controls 3.5, 2.0, respectively) 2.2 (1.5-3.4), p<0.001. Variables associated with mortality detected at any moment before death were kidney involvement 2.16 (1.09-4.29), p<0.02; the steroid therapeutic index 2.3 (1.2-4.5), p<0.001; number of previous admissions 2.4 (1.4-4.3), p<0.001; the MEX-SLEDAI index (cases 21.6 mu 6.3 sigma vs controls 12.6, 5.8), 1.2 (1.1-1.3), p<0.001; and the number of severe infections 14.4 (4.4-46.2), p<0.001. Protective variables were skin involvement 0.1 (0.3-0.6), p<0.001; daily dose of chloroquine (cases 3.9 mu, 24.1 sigma vs controls 39.4, 60.0 mg), p <0.0001 and the time from thefirst SLE symptom to the patient's demise or the cut-off date 0.7(0.6-0.9), p<0.001. Multiple logistic regression showed that the model which best explained mortality consisted of a severity index 2.6 (1.7-3.8), p<0.001; heart disease 6.5 (1.5-28.2), p=0.01, and steroid therapeutic index 3.3 (1.6-6.6), p=0.001. CONCLUSIONS: An active SLE with multi-organic involvement, steroids and infections were associated with mortality in Mexican patients with lupus attended in a tertiary care center A protective effect of cutaneous disease and chloroquine use was observed. PMID- 11491495 TI - Undifferentiated connective tissue disease with antibodies to Ro/SSa: clinical features and follow-up of 148 patients. AB - OBJECTIVE: To evaluate the clinical and serologic profile, the rate of progression to well defined CTD and the possible predictors of disease evolution in patients affected by UCTD with antibodies anti-RoISSA. METHODS: 148 patients diagnosed as UCTD were retrospectively evaluated. Antibodies to SSA/Ro were determined by counter-immunoelectrophoresis and ELISA. RESULTS: Thirty-six patients (24.3%) developed a well-defined CTD after a mean follow-up of 4.5 years. Most patients developed primary Sjogren's syndrome (SS) (50%) or systemic lupus erythematosus (SLE) (30.5%). Leukopenia and xerophthalmia developed more frequently in the group of patients evolving to defined CTDs (p < 0.0032 and p < 0.0063). Leukopenia independently predicted the evolution in CTD by multivariate regression analysis (p < 0.019). Anti-dsDNA predicted the evolution in SLE (p < 0.0207), while the presence of additional anti-ENA specificity to anti-Ro/SSA was not associated with the outcome. CONCLUSION: 24.3% of patients with UCTD and antibodies to Ro/SSA can progress in a relatively short period of time to well defined CTDs. The development of primary SS could be predicted by xerophthalmia and SLE by the appearance of anti-dsDNA antibodies. PMID- 11491496 TI - Circulating levels of Th1/Th2 cytokines in patients with primary Sjogren's syndrome: correlation with clinical and immunological features. AB - OBJECTIVE: To analyse the circulating levels of Th1 and Th2 cytokines in patients with primary Sjogren's syndrome (SS), as well as to investigate their association with clinical and immunological manifestations. METHODS: We included 62 consecutive patients (58 women and 4 men) seen in our Unit. All patients fulfilled 4 or more of the European diagnostic criteria for SS. Serum levels of IL-6 (pg/mL), IL-2 (pg/mL), srIL-2 (pM), TNFalpha (pg/mL) and IL-10 (pg/mL) were determined using a solid phase enzyme immunoassav performed on microtiter plate. RESULTS: When compared with the control group, high levels of Th1 (11-2, srIL-2) and Th2 (IL-6, IL-10) cytokines were detected in SS patients, although only IL-6 levels reached statistical significance. On the other hand, analysis of the mean serum concentrations of cytokines showed distinct patterns of elevated cytokines according to the organ involved, and elevated levels of IL-6 (126.5 v 20.6 pg/mL, p < 0.05) and IL-10 (10.6 v 2.2 pg/mL, p < 0.005) were observed in those patients with liver involvement. Analysis of the cytokine levels according to the presence of immunological features showed: higher levels of srIL-2 (95.6 v 54.0 pM, p < 0.05) in patients with anti-Ro/SS-A antibodies; increased levels of srIL-2 (111.4 v 59.4 pM, p < 0.05) in patients with antiLa/SS-B antibodies; higher levels of srIL-2 (90.4 vs 50.8 pM, p < 0.05) and TNFalpha (37.9 v 22.6 pg/mL, p = 0.001) in patients with RF and higher levels of IL-6 (88.0 v 23.1 pg/mL, p < 0.05) in patients with cryoglobulins and in those with hypocomplementemia (130.3 vs 21.0 pg/mL, p < 0.05). CONCLUSION: We found a significant elevation of several circulating cytokines in some clinical and immunological subsets of patients with primary SS. These cytokine patterns may be markers for specific extraglandular involvement in SS and could be of interest in assessing the response to treatment protocols or in monitoring the disease evolution. PMID- 11491497 TI - The effect of radiation synovectomy in patients with persistent arthritis: a prospective study. AB - OBJECTIVE: To investigate and compare the effects of radiation synovectomy of various joints in a rheumatological practice. METHODS: Consecutive patients referredfor radiation synovectomy to Medical Center Alkmaarfrom 1993 till 1996 were analyzed (n = 138). Patients had to have persistent arthritis despite at least two intra-articular glucocorticoid injections with 20 mg triamcinolone hexacetonide. The knees were treated with 185 MBq Yttrium-90; shoulders, elbows, wrists, hips and talocrural joints received 185 MBq Rhenium-186 and meta carpophalangeal joints and proximal interphalangeal joints 37 MBq Erbium-169. The radionuclide injection was followed by injection of 20 mg triamcinolone in order to prevent flare-up of synovitis (due to chemical irritation) and needle-track burn. The clinical effect was assessed by evaluating VAS pain (0-10 point scale), functional disability, tenderness and swelling of the treated joint and patient's and physician's global assessments of the effect of therapy (each on a 4-point scale). RESULTS: The overall success rate for radiation synovectomy one year after treatment was 70% with a significant improvement in the variables VAS pain, functional disability and joint tenderness and swelling, when compared to baseline values (p < 0.000001). Moderate to considerable satisfaction of patients and physicians one year after treatment was found for > 50% of cases. Wrists and shoulders were the joints with highest success rate of treatment, followed by the elbows. Lowest success rates were found for hips and ankles. In RA treatment was effective in 76% of cases whereas patients with OA exhibited a success rate of 50%. In RA all treated joints, except the ankles, exhibited a success rate of > or = 75%. No short-term clinical adverse side effects were noted. CONCLUSION: Radiation synovectomy seems to be a successful treatment for persistent arthritis when other therapeutic modalities have failed. PMID- 11491498 TI - Anti-beta2 glycoprotein I and anticardiolipin antibodies in leptospirosis, syphilis and Kala-azar. AB - OBJECTIVE: Reports have shown that anticardiolipin (aCL) antibodies present in patients with autoimmune diseases are dependent on the cofactor,beta2 glycoprotein I (beta2 GPI), as opposed to aCL antibodies seen in infectious diseases such as syphilis, HIV hepatitis C, etc. The assay for anti-beta2GPI antibodies has been reported to be more specific for antiphospholipid syndrome (APS). However, the prevalence of these antibodies in diseases such as leishmaniasis and leptospirosis remains unknown. The aim of the present study was determine the prevalence of antibodies to cardiolipin and to beta2GPI in patients with different infectious diseases, including leptospirosis, syphilis and leishmaniasis. METHODS: Samples from patients with Kala-azar (visceral leishmaniasis), syphilis or leptospirosis were tested for IgG and IgM anticardiolipin and IgG anti-beta2GPI antibodies by ELISA. RESULTS: In patients with Kala-azar the prevalence of IgG aCL, IgM aCL and anti-beta2GPI was 6% (2/30), 3% (1/30) and 53% (16/30), respectively. In syphilis the prevalence was 18% (14/74), 13% (10/74) and 10% (8/70), respectively. In leptospirosis the frequency of these antibodies was 23% (9/39), 10% (4/39) and 17% (6/34), respectively. There was no statistical correlation between aCL and anti-beta2GPI antibodies in these diseases. DISCUSSION: This study clearly shows a significant prevalence of anti-beta2GPI antibodies in leptospirosis and leishmaniasis and syphilis. This indicates that the assay for antibeta2GPI antibodies should be thoroughly validated before it is introduced as a definitive tool for the diagnosis of APS, testing a larger number of sera from patients with a wider range of clinical conditions. PMID- 11491499 TI - Glucocorticoid effects on myocardial performance in patients with systemic sclerosis. AB - OBJECTIVE: Myocardial inflammation andfibrosis are common autopsyfindings in systemic sclerosis (SSc) and, although symptomatic cardiac involvement occurs less often, current therapies remain empiric and do not prevent or modify its course. In this open, uncontrolled study we assessed the short-term effects of glucocorticoid administration on myocardial performance in patients with SSc in the absence of clinically overt cardiac disease. METHODS: Resting radionuclide ventriculography with 99mTc was performed before and 20 days after the administration of prednisolone, 20 mg daily, in 32 patients with SSc without clinically evident myocardial dysfunction at rest; 13 and 19 patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), respectively, were studied in parallel as controls. RESULTS: The mean left ventricular ejection fraction (LVEF) value at baseline was 59% in the SSc group; similar values were found for the SLE (61%) and RA (59%) groups. An impaired LVEF (i.e., <50%) was found in 6 patients with SSc and in 1 patient with SLE. Prednisolone administration resulted in a significant percent improvement in the baseline LVEF (mean 18%, p = 0.0001) in the SSc group; this improvement was greater in the patients with diffuse SSc than in those with limited skin disease (27% vs 10%, p = 0.02). The improvement was most prominent in the 6 patients with an initial impaired LVEF No significant improvement was observed in the SLE or RA control groups. The linear trend betveen the individual baseline LVEF values in patients with SSc and their percent changes after treatment (r2 = 0.55, p: 0.00001) showed that the lower the initial LVEF the greater the improvement caused by prednisolone. The degree of LVEF improvement was also associated with the individual erythrocvte sedimentation rate values and serum IgG concentrations at baseline. Prednisolone-induced changes in LVEF were not associated with any changes in blood pressure, heart rate, blood, plasma, or red cell volumes. CONCLUSION: Glucocorticoid administration may improve myocardial performance in some patients with SSc. Although further double-blind controlled studies of the long-term effects are warranted, such treatment may be useful in those patients with SSc and documented low LVEF if they are kept under careful observation for objective improvement. PMID- 11491500 TI - Immunogenetics and clinical aspects of Takayasu's arteritis patients in a Mexican Mestizo population. AB - OBJECTIVE: The aim of the present work was to study the association between HLA alleles and Takayasu's arteritis in Mexican Mestizo patients. METHODS: The study included 26 Mexican Mestizo patients with Takayasu's arteritis and 99 healthy unrelated individuals. HLA-A, -B and -DR alleles were determined by polymerase chain reaction PCR-SSP RESULTS: Increased gene frequencies were demonstrated for HLA-B15(p=0.009,pC=0.020,OR=3.24,EF=11.9%) and HLA-B52 (p=0.008, pC=0.027, OR=5.16, EF=7.7%), and a decreased frequency for the HLA-A24 allele in patients compared to normal controls (p=0.035, pC=NS, PF=11.1%). When HLA typing was correlated to clinicalfeatures in 24 cases, wefound an increasedfrequencies of HLA-DR14 in patients with systemic arterial hypertension (p=0.005, pC=0.004, OR=24.6, EF=38.3%) and HLA-A2 on patients with pulmonary involvement (p=0.034, pC=0.036, OR=3.67, EF=40.4%) when compared to patients without these clinical manifestations. CONCLUSION: These data confirm HLA-B52 as a relevant susceptibility allele for Takayasu's arteritis and suggest that HLA-B15 could be important as a marker of the disease in Mexican patients. Other class I and/or class II alleles could also be relevant as markers for the clinical features present in these patients. PMID- 11491501 TI - Focal sialadenitis in patients with early synovitis. AB - OBJECTIVE: To investigate the frequency of sialadenitis on lip biopsy in patients with synovitis of recent onset (ES), and see how sialadenitis relates to clinical and laborator findings of ES. METHODS: Joint involvement, laboratory measures and biopsies of the minor salivary glands were evaluated in 10 ES patients. Diagnosis at a one-year follow-up exam was noted. RESULTS: Six ES patients (60%) had a positive lip biopsy (mononuclear cell focus score greater than 1). ES patients with a positive lip biopsy presented with oligoarthritis, while ES patients with a negative lip biopsy had a more polyarticular presentation. No differences in laboratory measures between patients with a positive and negative lip biopsy were present. Seven ES patients had a diagnosis of rheumatoid arthritis and three had undifferentiated arthritis at the end of one year. CONCLUSION: ES patients had a higher than expected frequency offocal sialadenitis. PMID- 11491502 TI - A comparative study of outcome in myositis and other musculoskeletal disorders assessed using the Nottingham health profile. AB - OBJECTIVE: This study evaluated the comparative impact of myositis and other musculoskeletal disorders on general health using the Nottingham health profile (NHP) as a generic measure of health status. METHODS: A prospective observational study of 113 females with myositis, 142 females with rheumatoid arthritis, 45 females with spinal osteoporosis and 96 females with knee osteoarthritis. RESULTS: All mean NHP section scores were higher in myositis and other musculoskeletal disorders compared to population mean values. Section scores for energy and social isolation were high in myositis compared to all other disorders. Scores for physical disability in myositis were similar to RA. Pain scores were higher in RA and OA compared to myositis. Backwards linear regression models explained 26-42% of the variation in energy and social isolation scores. Emotion and physical section scores were the major determinants and the pattern was similar in all disorders. Disease duration and age had little effect. CONCLUSIONS: Myositis is not simply a disease with physical problems but has wide ranging effects on social and emotional well being. Until disease-specific instruments are available, a generic measure like the NHP can be used to assess problems other than muscle pain and loss of strength. PMID- 11491503 TI - Microsatellite polymorphism of the MICA gene and susceptibility to rheumatoid arthritis. AB - We examined the role of MICA gene alleles in susceptibility to rheumatoid arthritis (RA). Ninety adult Caucasian patients with classical seropositive RA and 85 normal healthy Caucasian subjects from the same geographical area were typed for microsatellite repeat polymorphism in the transmembrane region of the MICA gene by the polymerase chain reaction. The results show that the MICA allele 6 may confer protection from the development of RA. PMID- 11491504 TI - Surface EMG of the lumbar part of the erector trunci muscle in patients with fibromyalgia. AB - OBJECTIVE: To determine differences supposed in EMG parameters of the erector trunci region between patients with fibromyalgia and healthy subjects during defined investigation situations. METHODS: During sitting and standing in upright position surface EMG (SEMG) from 15 subjects with fibromyalgia and 10 healthy controls was performed using a 16-channel technique where the electrodes were applied in a well-defined grid pattern (gain 5000, 3 db points at 5 Hz and 700 Hz respectively). SEMG quantification was done by Fourier algorithm using 512 measurement points for calculation. RESULTS: An increased EMG amplitude could be recorded during rest in fibromyalgia patients compared with controls. Spatial amplitude differences (frequency range 100-500 Hz) in the low back region were significantly (p < 0.01) decreased in the patients' group during sitting. CONCLUSION: It is the first time that a decreased difference in EMG amplitude of different parts within a certain muscle could be proven in patients with fibromyalgia. As far as is known from the literature this result seems to be a uniquefinding in fibromyalgia patients. PMID- 11491505 TI - Hypercapnic coma due to diaphragmatic involvement in a patient with dermatomyositis. AB - We report an acute respiratory distress by diaphragmatic involvement due to dermatomyositis. A fifty year-old patient with typical dermatomyositis presented an acute respiratory insufficiency with hypercapnic coma due to diaphragmatic muscle involvement. Respiratory state required mechanical ventilation initially and improved secondarily gradually upon corticosteroids and intravenous immunoglobulins. Only few cases of acute respiratory distress in dermatomyositis due to respiratory muscle involvement are reported in literature. PMID- 11491506 TI - Widespread nocardiosis in two patients with Behcet's disease. AB - Nocardia spp., a group of gram-positive variably acid-fast aerobic bacteria, are opportunistic pathogens in immunocompromised hosts. We here-within describe 2 cases of widespread nocardiosis in patients with Behcet's disease. In addition to endogen endophthalmitis in case 1, both cases developed lung, brain and skin involvement. Despite brain involvement, the prognosis was good, although vision was completely lost in case 1, which was directly attributable to a delay in both diagnosis and treatment. PMID- 11491507 TI - Cogan's syndrome: organ-specific autoimmune disease or systemic vasculitis? A report of two cases and review of the literature. AB - Cogan's syndrome is a rare disorder of unknown origin characterized by inflammatory eye disease and vestibuloauditory symptoms. Typically, young adults suffer from interstitial keratitis and sudden onset of tinnitus and hearing loss. Few cases (around 150) have been published and thus it is difficult to determine the percentage of patients with underlying systemic disorders such as systemic vasculitis. The variety of systemic manifestations is large and includes fever, splenomegaly, lymphadenopath, and musculoskeletal complaints. Systemic vasculitis can be seen in around 10% of cases and may involve the large vessels, appearing as Takayasu-like vasculitis with affection of the aortic valve but also the coronary arteries and the small kidney vasculature. Evaluating the exact extension of the systemic features determines the choice of treatment. While corticosteroids have proved to be of short-term benefit, long-term treatment with immunosuppressive drugs is controversial. Auditory function in deaf patients has often been restored successfully with cochlear implants. To illustrate the nature of the syndrome, we present two patients with a wide clinical spectrum of symptoms from local disease restricted to the eyes and ears to a widespread vasculitis affecting arteries of the brain, kidney and the upper and lower extremities. We then review the typical aspects as well as the etiology of the disease. PMID- 11491508 TI - Anti-keratin antibodies in patients with juvenile idiopathic arthritis. AB - OBJECTIVE: We discuss the presence of anti-keratin antibodies (AKA) of the IgG class in patients with defined juvenile idiopathic arthritis (JIA). METHODS: An indirect immunofluorescence test with rat oesophagus substrate was used for the detection and quantification of AKA antibodies in patients'sera. RESULTS: Overall 30/60 patients with JIA had sera positiveforAKA (50%, p=0.0005) ranging from 1:20 to 1:160 dilutions. Using the classification criteria for childhood idiopathic arthritis, AKA occurred in 2/7 patients with systemic disease (28.6%), in 13/30 patients with RF negative polyarthritis (43.3%, p=0.008) and in 12/18 RF positive polyarthritis (66.7%, p=0.002). AKA were also found in a small cohort of patients with oligoarthritis (1/3) and psoriatic arthritis (2/2). AKA positivity occurred in 3/26 healthy controls at a 1:20 dilution. The presence ofAKA was correlated as well as with the severity of the disease. Our study revealed that AKA was present overall in 16/29 patients (55.2%) with severe JIA and in 11/26 patients (42.3%) with non-severe disease. We also observed that AKA remained positive regardless of disease activity. AKA were detectable in 44.4% patients with active JIA and in 45.9% patients in the complete or near remission. CONCLUSION: Our data suggest that AKA are present in patients with JIA. However no correlation with severity or disease activity was observed. PMID- 11491509 TI - Uncommon causes of liver disease in juvenile systemic lupus erythematosus. PMID- 11491510 TI - A case report of a man with rheumatoid factor positive rheumatoid arthritis associated with collagenous colitis. PMID- 11491511 TI - Apoptotic neutrophils and anticardiolipin antibodies. PMID- 11491512 TI - Beneficial effects of leflunomide in glucocorticoid- and methotrexate-resistant Takayasu's arteritis. PMID- 11491513 TI - Blue coloured skin in psoriatic arthritis. PMID- 11491514 TI - Giant cell arteritis presenting as occipital neuralgia. PMID- 11491515 TI - Sarcoidosis presenting as Heerfordt's syndrome with myopathy. PMID- 11491516 TI - Spondyloepiphyseal dysplasia tarda in a child with severe and an adult with mild clinical features. PMID- 11491517 TI - Pachydermodactily as a cause of painful swelling of the knuckles: successful treatment with intralesional steroids. PMID- 11491518 TI - Pneumatosis intestinalis in a patient with polymyositis. PMID- 11491519 TI - Sclerodermic lesions after liposuction in a patient with Raynaud's phenomenon and anti-centromeric antibodies. PMID- 11491520 TI - Treatment of TRAPS with etanercept: use in pediatrics. PMID- 11491521 TI - Bioaccumulation of yttrium in Pseudomonas fluorescens and the role of the outer membrane component(s). AB - Pseudomonas fluorescens was grown in millimolar amounts of yttrium. The tolerance to yttrium appeared to be mediated by the ability of the organism to accumulate the trivalent metal predominantly in the outer membrane component(s). At the stationary phase of growth, 65 to 70% of the metal was associated with the constituent(s) of the outer membrane. Treatment with 2 mM (EDTA) did not release the metal. Incubation of the outer membrane fraction with yttrium led to further accumulation of the metal. The outer membrane equivalent to 1 mg of protein was shown to immobilize 175 microg of yttrium. There was no significant variation in uronic acid and the lipid contents of the control and yttrium-stressed cells as monitored by colorimetric assays. The protein profiles of the outer and inner membrane components obtained from the control and metal-stressed cells showed marked variations as revealed by sodium dodecyl sulphate polyacrylamide gel electrophoretic analysis. PMID- 11491522 TI - Correlation between Congo red binding and contact haemolysin production in Shigella species. AB - Haemolytic strains of Shigella dysenteriae type 1, Shigella flexneri, Shigella boydii and Shigella sonnei cultured on Congo red agar produced pigmented colonies (Pcr+) whereas nonhaemolytic strains produced white colonies and did not bind Congo red (Pcr-). S. flexneri-1 haemolysin negative mutant (lacking plasmid) of haemolysin positive prototroph also did not bind Congo red and produced nonpigmented colonies. Among the twelve strains of Shigella included in this study, the characteristics of Congo red binding, plasmid profile and haemolytic activity appeared to be correlated. Congo red binding occurred comparatively more by haemolysin-producing strains. Congo red binding can be used as a quick and reliable method for virulence traits of pathogens, including haemolysin activity. PMID- 11491523 TI - Microcosm studies on the persistence of peanut inoculant in acid soil and its interaction with indigenous soil micro-organisms. AB - The effect of a native peanut (Arachis hypogaea) microsymbiont strain (streptomycin resistant mutant) introduced into acid soil on the indigenous micro organism population as well as the survival of this isolate was studied. Although the isolate survived at a fairly low population level after the 3 month period of soil incubation, it maintained its antibiotic resistance. Slight changes in the total bacteria and fungi soil populations as a result of this inoculation were also detected. PMID- 11491524 TI - An evaluation of a polyantigenic ELISA to detect Epstein-Barr virus reactivation. AB - The diagnostic reliability of the Enzygnost EBV test (DadeBehring, Germany) for the detection of IgG and IgA antibodies in the diagnosis of Epstein-Barr virus (EBV) recurrent disease was investigated. Of 81 serum samples examined there were fourteen asymptomatic patients without EBV infection, 46 with past EBV infection, and 21 patients with EBV reactivation. The Enzygnost EBV test was based on an enzyme-linked immunosorbent assay with a pool of viral antigens. The reliability of IgG at >650 IU/ml, and IgA for the diagnosis of reactivation or chronic persistent EBV infection gave 100% sensitivity, 83.3% and 98.3% specificity, respectively. The data indicated that the appearance of EBV IgA was associated with EBV reactivation together with clinical manifestations. PMID- 11491525 TI - Mycolic acids from Mycobacterium tuberculosis: purification by countercurrent distribution and T-cell stimulation. AB - Bacterial cell wall lipids are recognized as immunostimulatory molecules which make an important component of vaccines against bacterial diseases. Even mycolic acids, forming the waxy outer layer of the bacilli which cause tuberculosis, have been shown to stimulate human CD4/8 double negative T-cells. The role of these cells in resistance to tuberculosis is currently still debated. In this work, a method is described to purify mycolic acids from bacterial crude extracts in a single step using countercurrent distribution. Mycolic acids obtained in this way approach 100% purity and stimulate both double negative and CD4 positive T-cells in peripheral blood leucocytes obtained from healthy human donors. Stimulation of CD4 cells by mycolic acid antigens has not been reported before, emphasizing the potential importance of mycolic acids in the context of the fight against tuberculosis. PMID- 11491526 TI - Enzymatic activity of micro-organisms isolated from cork wine stoppers. AB - The production of enzymes by micro-organisms which are found on vegetal substrates is important due to their ability to decompose cellulose, lignin and other components, which guarantee the integrity of the vegetal cell. The objective of this study was to determine the enzymatic activity of filamentous fungi, yeasts and bacteria, isolated from natural cork stoppers for bottles of still and sparkling wines. Suspensions of fungal conidia, yeasts and bacterial cells of micro-organisms were established in concentrations of 10(6) CFU/ml. The enzymatic activity of these micro-organisms was evaluated by means of the API ZYM system, with which it was possible to determine and semi-quantify nineteen enzymatic activities simultaneously. The enzymes produced by all of the species were esterase (C1), esterase lipase and naphthol-AS-BI-phosphohydrolase. The micro-organisms with the greatest enzymatic activity were Monilia sitophila, Alternaria alternata, Aspergillus niger and Aeromonas sp. PMID- 11491527 TI - Polyamine distribution profiles in the eighteen genera phylogenetically located within the Flavobacterium-Flexibacter-Cytophaga complex. AB - Cellular polyamines of eighteen genera belonging to the Flavobacterium Flexibacter-Cytophaga complex were analysed by ion exchange liquid chromatography. Homospermidine was the major polyamine in the genera Bergeyella, Riemerella, Ornithobacterium, Weeksella, Capnocytophaga, Polaribacter and Psychroflexus belonging to the family Flavobacteriaceae. In the family Spirosomaceae, Runella, Spirosoma and Flectobacillus species contained spermidine whereas Cyclobacterium species contained homospermidine. Within a divergent cluster, Haliscomenobacter and Lewinella species contained spermidine whereas Saprospira grandis contained agmatine alone. The major polyamine of Chitinophaga and Sporocytophaga species was homospermidine. Flexithrix dorotheae contained spermidine. Microscilla marina, the type species of the genus Microscilla, contained spermidine and cadaverine. However, 'Microscilla sericea' contained homospermidine, 'Microscilla furvescens' contained spermidine, and 'Microscilla arenaria' lacked all polyamines. Polyamine profiles serve as a phenotypic chemotaxonomic marker for the reclassification of the genera belonging to the complex. PMID- 11491528 TI - HLA expression in cancer: implications for T cell-based immunotherapy. AB - HLA class I expression is altered in a significant fraction of the tumor types reviewed here, reflecting either immune pressure or, simply, the accumulation of pathological changes and alterations. However, in all tumor types analyzed, a majority of the tumors express HLA class I. with a general tendency for the more severe alterations to be found in later-stage and less differentiated tumors. These results are encouraging for the development of specific immunotherapies, especially considering that (1) the relatively low sensitivity of immunohistochemical techniques might underestimate HLA expression in tumors, (2) class I expression can be induced in tumor cells as a result of local inflammation and lymphokine release, and (3) class I-negative cells would be predicted to be sensitive to Iysis by natural killer cells. PMID- 11491529 TI - A novel polymorphism in the 5' promoter region of the human interleukin-4 receptor alpha-chain gene is associated with decreased soluble interleukin-4 receptor protein levels. AB - Interleukin (IL)-4 exerts its biological effects through binding to the IL-4 receptor (IL4R) complex, plays a central role in stimulating B-cell differentiation, and is crucial for the development of T helper 2 cells. Recently, a soluble form of the human IL4R alpha chain (sIL4R alpha), which is produced by alternate mRNA splicing of exon 8, was discovered. sIL4R is thought to play an important role in either enhancing or inhibiting IL-4 signalling. We analyzed the 5' promoter region of the human IL4R alpha-chain gene (IL4RA) of healthy volunteers by DNA sequencing and found three novel single-nucleotide polymorphisms (SNPs; T-890C, T-1914C, C-3223T) and one novel short tandem repeat [(CAAAA)(5-7)-3600]. The two common promoter region SNPs T-1914C and C-3223T as well as six known coding SNPs in the IL4RA gene were genotyped in healthy blood donors by PCR with sequence-specific primers; total sIL4R levels were measured by ELISA. Results revealed a highly significant association of the -3223T variant with lowered sIL4R levels (two-tailed t-test, P=0.0002). Results remained highly significant after Bonferroni adjustment for multiple comparisons (P=0.0017). Moreover, the C-3223T variant was found to be in strong linkage disequilibrium with the extracellular 150V variant (P<0.001), which was recently described to be associated with atopic asthma in a Japanese population. Since this novel IL4RA promoter region SNP is common (allele frequency 29.8%), we conclude that it may be of importance for the genetic regulation of the IL-4 signalling pathway. PMID- 11491530 TI - Conserved organization of the ILT/LIR gene family within the polymorphic human leukocyte receptor complex. AB - The human leukocyte receptor complex (LRC) at Chromosome 19q13.4 encodes Ig superfamily proteins which regulate the function of various hematopoietic cell types. We investigated characteristics of the Ig-like transcript (ILT)/leukocyte Ig-like receptor (LIR) group of LRC genes in comparison with the other major LRC loci encoding the killer cell Ig-like receptors (KIRs). In direct contrast to KIR genes, the ILT/LIR loci of ethnically diverse individuals did not display haplotypic variations in gene number. Investigation of gene expression identified novel cDNA sequences related to the ILT2/LIR1, ILT4/LIR2, ILT3/LIR5, and ILT7 loci, while phylogenetic analysis revealed two distinct lineages of ILT/LIR genes. These two lineages differ in both the nature and extent of their sequence polymorphism. The presence of certain transcription factor-related motifs in the 5' untranslated region of ILT/LIR cDNAs correlates with the specific cell types in which particular ILT/LIR genes are expressed. Although extensive gene duplications and conversion events have apparently forged the LRC, our results indicate striking conservation in the organization of the ILT/LIR genes when compared with the related and closely linked KIR genes. This suggests the evolutionary maintenance of a significant function consistent with the cellular distribution of the ILT/LIR proteins. PMID- 11491531 TI - Interactions of human NKG2D with its ligands MICA, MICB, and homologs of the mouse RAE-1 protein family. AB - NKG2D is an activating receptor that is expressed on most natural killer (NK) cells, CD8 alphabeta T cells, and gammadelta T cells. Among its ligands is the distant major histocompatibility complex class I homolog MICA, which has no function in antigen presentation but is induced by cellular stress. To extend previous functional evidence, the NKG2D-MICA interaction was studied in isolation. NKG2D homodimers formed stable complexes with monomeric MICA in solution, demonstrating that no other components were required to facilitate this interaction. MICA glycosylation was not essential but enhanced complex formation. Soluble NKG2D also bound to cell surface MICB, which has structural and functional properties similar to those of MICA. Moreover, NKG2D stably interacted with surface molecules encoded by three newly identified cDNA sequences (N2DL-1, 2, and -3), which are identical to the human ULBP proteins and may represent homologs of the mouse retinoic acid-early inducible family of NKG2D ligands. Because of the substantial sequence divergence among these molecules, these results indicated promiscuous modes of receptor binding. Comparison of allelic variants of MICA revealed large differences in NKG2D binding that were associated with a single amino acid substitution at position 129 in the alpha2 domain. Varying affinities of MICA alleles for NKG2D may affect thresholds of NK-cell triggering and T-cell modulation. PMID- 11491532 TI - Cloning of human DECTIN-1, a novel C-type lectin-like receptor gene expressed on dendritic cells. AB - We identified a new Ca2+-dependent lectin-like receptor gene, DECTIN-1 (HGMW approved symbol CLECSF12), the human orthologue of mouse Dectin-1, coding for a putative type II transmembrane glycoprotein with an extracellular C-type lectin like domain. The gene structure and two alternative spliced forms of DECTIN-1 are described. The DECTIN-1 gene was localized in the natural killer gene complex on human Chromosome 12p12.3-p13.2, between OLR1 and CD94 (position 21.8 cM on the genetic map). The DECTIN-1 gene is highly expressed at the mRNA level in dendritic cells and is not further up-regulated during the maturation of these cells with tumor necrosis factor-alpha. The DECTIN-1 gene therefore represents a novel human member of the C-type lectin-like receptor gene family preferentially expressed in dendritic cells. PMID- 11491533 TI - Vanin genes are clustered (human 6q22-24 and mouse 10A2B1) and encode isoforms of pantetheinase ectoenzymes. AB - The mouse Vanin-1 molecule plays a role in thymic reconstitution following damage by irradiation. We recently demonstrated that it is a membrane pantetheinase (EC 3.56.1.-). This molecule is the prototypic member of a larger Vanin family encoded by at least two mouse (Vanin-1 and Vanin-3) and three human (VNN1, VNN2, VNN3) orthologous genes. We now report (1) the structural characterization of the human and mouse Vanin genes and their organization in clusters on the 6q22-24 and 10A2B1 chromosomes, respectively; (2) identification of the human VNN3 gene and the demonstration that the mouse Vanin-3 molecule is secreted by cells, and (3) that the Vanin genes encode different isoforms of the mammalian pantetheinase activity. Thus, the Vanin family represents a novel class of secreted or membrane associated ectoenzymes. We discuss here their possible role in processes pertaining to tissue repair in the context of oxidative stress. PMID- 11491534 TI - R-phycoerythrin-cyanine 5 tandem discerns CD72 polymorphism. AB - By fluorescence analysis, we demonstrated that a fluorochrome tandem composed of R-phycoerythrin and cyanine 5 specifically recognized B cells from SJL, AKR, MRL/Mp, and NOD mouse strains, whereas B cells from C57BL/6, DBA/2, SWR, 129/Sv, and BALB/c were not stained. A strict correlation was observed between the fixation of the fluorochrome and the pattern of expression of the pan-B cell marker CD72, i.e., early expression in B-cell lineage development and downregulation on the terminally differentiated activated B cells. Three allelic forms, CD72a, CD72b and CD72c have been well characterized, and show a high number of amino acid substitutions concentrated in the membrane-distal extracellular domain. Using a PCR approach, we determined that all mouse strains positive for fluorochrome staining display the CD72c allelic form. Moreover, a genetic analysis showed that the fixation of the fluorochrome on the B cell is exclusively dependent on the presence of the CD72c allele. Together, these results strongly suggest that the tandem binds a molecular complex comprising the CD72c molecule or recognizes directly the CD72c molecule itself. PMID- 11491535 TI - Cloning, sequencing, and homology analysis of nonhuman primate Fas/Fas-ligand and co-stimulatory molecules. AB - The finding that a single administration of select recombinant human cytokines to nonhuman primates leads to potent cytokine-neutralizing antibody responses in the heterologous host despite >95% homology at the nucleotide and protein level prompted our laboratory to clone, sequence, and prepare recombinant nonhuman primate cytokines, chemokines, growth factors, and other immunoregulatory molecules. In the present report, we present findings on the gene sequences encoding the nonhuman primate homologues of human CD80, CD86, their ligands CD28 and CD152, CD154, CD95, and CD95-L from rhesus macaques and for phylogenetic analysis from pig-tailed macaques, African sooty mangabey monkeys, baboons, and vervets as well as select molecules from the New World aotus and marmoset monkeys. With the exception of CD95, the homology between nonhuman primate and human co-stimulatory molecules was above 95%. In contrast, CD95 was only 89.2% homologous to human CD95, but the differences were essentially found in the transmembrane and intracellular (death) domains. The extracellular portion of CD95 was more homologous which was in accordance with approximately 98% homology between Old World monkey and human CD95-L. In general, sequences from the New World monkey species appeared equidistant to sequences from Old World species and humans in terms of homology suggesting distinct evolutionary patterns. Of interest was the isolation of various splice variants of monkey CD86, CD152 (CTLA 4), CD154, and CD95 transcripts. This is also the first report documenting the occurrence of natural CD86 variants with deleted transmembrane domains, found both in sooty mangabeys and baboon RNA samples. Monkey CD95 showed various deletions and addition of residues in the transmembrane and intracytoplasmic domains compared with human CD95 and between Old and New World species. Subcloning of rhesus CD154 into an expression vector demonstrated expression of a functional protein in cell culture. The other genes are being cloned into expression vectors for the preparation and biological characterization of the nonhuman primate molecules. These investigations will provide novel reagents for in vivo use as immunomodulatory reagents in nonhuman primates in studies which may provide a rationale for their use in humans. PMID- 11491536 TI - Allelic polymorphism in MHC class II B in four populations of Atlantic salmon (Salmo salar). AB - We sequenced exon 2 of the MHC class II B gene in Atlantic salmon from the Baltic Sea and identified 17 different exon 2 alleles among 22 different restriction fragment length polymorphism haplotypes. The sequences differed at between 1 and 34 bases. Two different tests were used to estimate the importance of recombination in the generation of new alleles. Recombination events appear to have occurred between three and nine times. Only two pairs of sequences differed by less than five nucleotides, minimizing the importance of point mutations for generating new alleles. Phylogenetic analysis showed that sequences did not cluster according to populations, and genetic distances between populations were small compared to those obtained by allele frequency data. These results, together with the similarity found between exon 2 sequences from Baltic salmon and Norwegian salmon, indicate that all of the identified alleles were present in the ancient salmon population colonizing the Baltic rivers after the last glaciation. PMID- 11491537 TI - Complete sequence, genomic organization, and chromosomal localization of the human gene encoding the SHP2-interacting transmembrane adaptor protein (SIT). PMID- 11491538 TI - Multiple species express thiol oxidoreductases related to GILT. PMID- 11491539 TI - The mouse Dap10 gene. PMID- 11491540 TI - Detection of transcripts for a soluble form of the RT1-E MHC class Ib molecule in rat placenta. PMID- 11491541 TI - Paleoecotoxicology: extending environmental toxicology and chemistry to the interpretation of the fossil record. PMID- 11491542 TI - Measurement of Henry's law constant for methyl tert-butyl ether using solid-phase microextraction. AB - Increasing groundwater contamination with methyl tert-butyl ether (MTBE) requires more efficient remediation technologies. Accurate measurement of MTBE's air-water partitioning coefficient (Henry's law constant, H) is important for the design and optimization of removal efficiency for many treatment systems as well as for predicting its fate and transport. Previously published data for MTBE appear to have some unusual nonlinearity at lower temperatures (15-30 degrees C), and a wide range of values exists for dimensionless H at 25 degrees C, from 0.0216 to 0.1226 in the published literature. We measured H for MTBE using headspace solid phase microextraction (SPME) and a static method that considers equilibrium partitioning in a closed system, for temperatures between 15 and 4 degrees C. To validate our methods, we measured H for benzene, toluene, and trichloroethylene and compared our results to previously published values, with excellent agreement. The Arrhenius plot for MTBE indicates that ln(HMBTE) = 6.85 - 2,900 T( 1), with T in K. At 25 degrees C, HMBTE = 0.0555 +/- 0.0122. PMID- 11491543 TI - The relationship of Chironomus riparius larval Se body burden and body concentration to larval dry mass and effects on sensitivity to selenium. AB - Selenium as selenite was added to laboratory cultures of Chironomus riparius. In two sets of cultures, substrate-bound Se concentrations were typically 10(3) times dissolved Se concentrations, while in the other set dissolved Se concentrations were about 10 to 50 times substrate-bound Se concentrations. Body burdens of individual second-, third-, and fourth-instar larvae and eviscerated fourth-instar larvae were measured. Regressions of Se body burden on larval dry mass found evidence for separate intercepts but common slopes for each instar, and Se body concentrations were diluted by growth. The derivation of body concentration from body burden as a function of larval dry mass was used to derive a model of body burden and body concentration as a function of instar. Toxicity tests were performed to check for a change in sensitivity to Se as a function of culture Se concentration. Larval Se concentrations were highly correlated with dissolved Se concentrations but not with substrate-bound Se concentrations. Increases in Se in substrate caused no sensitization, while increases in dissolved Se caused desensitization. We concluded that covariates such as larval dry mass should be incorporated when deriving water quality criteria. PMID- 11491544 TI - Dose- and time-dependent formation of biliary benzo[a]pyrene metabolites in the marine flatfish dab (Limanda limanda). AB - Polycyclic aromatic hydrocarbons (PAHs) are abundant pollutants, and many PAHs are carcinogenic, but only after metabolic activation. Benzo[a]pyrene (BaP) is among the most carcinogenic PAHs. The dose and time response of two enzymes involved in BaP metabolism and the amounts of BaP metabolites excreted into the bile were evaluated in an experiment with dab (Limanda limanda). Ninety dab were exposed orally to one of five doses of BaP (0, 0.08, 0.4, 2, or 10 mg/kg) and sampled at 3, 6, or 12 d after exposure. None of the doses studied caused significant induction of either microsomal ethoxyresorufin-O-deethylase (EROD). which reflects cytochrome P450 1A (CYP1A) activity, or cytosolic glutathione-S transferase activity (GST). Concentrations of biliary BaP metabolites significantly increased with dose and significantly decreased with time after exposure. It is concluded that biliary BaP metabolites provide a much more sensitive method than EROD (CYP1A) or GST activity to monitor recent exposure to PAHs in dab. PMID- 11491545 TI - Determination of microcystins in cyanobacterial blooms by solid-phase microextraction-high-performance liquid chromatography. AB - A solid-phase microextraction (SPME) method was applied to study microcystin (MC) profiles in a natural Microcystis sp. bloom in a freshwater pond in Guangzhou, China. Three dominant MC variants, namely MC-LR, MC-YR, and MC-RR, were quantified. Simultaneous study of their total, extracellular, and intracellular profiles was made possible using SPME coupled to high-performance liquid chromatography. The total and intracellular concentrations of MC-LR in the bloom were 8.67 x 10(-2) microg/ ml and 1.93 mg/g, respectively. The corresponding concentrations of MC-YR were 1.20 x 10(-3) microg/ml and 0.06 mg/g, respectively, and those of MC-RR were 5.57 x 10(-2) microg/ml and 1.49 mg/g, respectively. Only MC-LR was detectable in the extracellular phase (1.49 x 10(-2) microg/ml) of the bloom, and its concentration was 14% of the intracellular content. Mass balance consideration revealed that only 71.1% of total MC-LR, 36.0% of total MC-YR, and 67.4% of total MC-RR within the cyanobacterial cells were released into the aqueous phase immediately after cell lysis. PMID- 11491546 TI - Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 1. Short-term effects on soil microbiology. AB - Linear alkylbenzene sulfonates (LAS) may occur in sewage sludge that is applied to agricultural soil, in which LAS can be inhibitory to biological activity. As a part of a broader risk assessment of LAS in the terrestrial environment, we tested the short-term effects of aqueous LAS on microbial parameters in a sandy agricultural soil that was incubated for up to 11 d. The assays included 10 microbial soil parameters; ethylene degradation; potential ammonium oxidation; potential dehydrogenase activity; beta-glucosidase activity; iron reduction; the populations of cellulolytic bacteria, fungi and actinomycetes; the basal soil respiration; and the phospholipid fatty acid (PLFA) content. Except for beta glucosidase activity, basal respiration, and total PLFA content, all soil parameters were sensitive to LAS, with EC10 values in the range of less than 8 to 22 mg/kg dry weight. This probably reflected a similar mode of LAS toxicity, ascribed to cell membrane interactions, and showed that sensitivity to LAS was common for various soil microorganisms. The extracellular beta-glucosidase activity was rather insensitive to LAS (ECI10, 47 mg/kg dry wt), whereas the basal soil respiration was not inhibited even at 793 mg/kg dry weight. This was interpreted as a combined response of inhibited and stimulated compartments of the microbial community. The PLFA content, surprisingly, showed no decrease even at 488 mg/kg. In conclusion, LAS inhibited specific microbial activities, although this could not be deduced from the basal respiration or the total PLFA content. The lowest EC10 values for microbial soil parameters were slightly higher than the predicted no-effect concentrations recently derived for plants and soil fauna (approximately 5 mg/kg dry wt). PMID- 11491547 TI - Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 2. Effects on soil microbiology as influenced by sewage sludge and incubation time. AB - The anionic surfactant linear alkylbenzene sulfonate (LAS) may inhibit soil microorganisms and may occur in agricultural soil through the application of sewage sludge. For five microbial parameters (microbial biomass C and the potentials of iron reduction, ammonium oxidation, dehydrogenase activity, and arylsulfatase activity), we compared the effects of aqueous LAS and LAS-spiked sewage sludge added to existing levels of 0, 3, 8, 22, 22, 62, 174, and 488 mg/kg soil (dry wt) in a Danish sandy agricultural soil that was incubated for 5 d to eight weeks. Arylsulfatase activity (measured after four weeks of incubation) was rather insensitive to LAS, with an EC 10 of 222 and more than 488 mg/kg in soil samples treated with aqueous LAS and LAS-spiked sewage sludge, respectively. For the other microbial parameters, the short-term effects (approximately one to two weeks) of aqueous LAS were characterized by an EC10 in the range of 3 to 39 mg/kg. Application of LAS via sewage sludge generally reduced the short-term effects for the microbial parameters, and the EC10 for LAS in sludge-amended soil after approximately one to two weeks of incubation ranged from less than 8 to 102 mg/kg. Recovery potential was seen for most microbial parameters as a result of prolonged incubation, both under conditions of LAS persistence (anaerobic conditions, the iron-reduction test) and LAS depletion (aerobic incubations, all other assays). In conclusion, the short-term inhibitory effects of LAS on soil microbiology were decreased in the presence of sewage sludge and by a prolonged (two to eight weeks) laboratory incubation period. PMID- 11491548 TI - Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 3. Sublethal effects on soil invertebrates. AB - Sewage sludge applied to agricultural soils often contains considerable amounts of linear alkylbenzene sulfonates (LAS). Toxic effects of LAS on soil organisms should, therefore. be evaluated to ensure safe use of sewage sludge as a fertilizer. In this study, dose-response relationships for the toxicity of Na-LAS to six species of soil invertebrates (survival, reproduction, and growth) were established using a sandy, agricultural soil as test substrate. In general, toxic effects on reproduction and growth appeared when the concentration in soil exceeded 40 to 60 mg/kg. Reproduction was approximately fourfold more sensitive in earthworms and enchytracids than in springtails and mites. It is argued that this difference in sensitivity is related to the dependency of soil pore water, which is high in the annelids but comparatively low in the arthropods. PMID- 11491549 TI - Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 4. The influence of salt speciation, soil type, and sewage sludge on toxicity using the collembolan Folsomia fimetaria and the earthworm Aporrectodea caliginosa as test organisms. AB - Sewage sludge applied to agricultural soils often contains linear alkylbenzene sulfonates (LAS) in the range of 1 to 10 g/kg dry weight, and their toxicity to relevant soil organisms should, therefore, be assessed to ensure safe use of sewage sludge as a fertilizer. Studies of LAS toxicity to soil organisms are few, and to our knowledge, factors that may influence the toxicity in the field have not been studied in detail. In this paper, we report on the influence of speciation of LAS in the test solution added to soil (soluble Na-LAS vs poorly soluble Ca-LAS or Mg-LAS), the influence of soil type, and the modifying effects of sludge amendment on the toxicity of LAS. These issues were investigated using reproduction of Collembola and growth of juvenile earthworms as test parameters. Speciation of the LAS added to test soil did not have any influence on toxicity for any of the test species. Likewise, in three different agricultural soils (sand, loam, and clay), we found almost equal toxicities. The LAS added to test soil in a sludge-water suspension was equally toxic as when it was added in an aqueous solution. However, anaerobic incubation for 7 and 14 d of the LAS-sludge suspension (with no decay of LAS) caused the toxicity to increase almost threefold in both collembolan and earthworm. The relationships between soil constituents, bioavailability, and toxicity are also discussed. PMID- 11491550 TI - Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 5. Probabilistic risk assessment of linear alkylbenzene sulfonates in sludge-amended soils. AB - Linear alkylbenzene sulfonates (LAS) can be found in high concentrations in sewage sludge and, hence, may enter the soil compartment as a result of sludge application. Here, LAS may pose a risk for soil-dwelling organisms. In the present probabilistic risk assessment, statistical extrapolation has been used to assess the risk of LAS to soil ecosystems. By use of a log-normal distribution model, the predicted no-effect concentration (PNEC) was estimated for soil fauna, plants, and a combination of these. Due to the heterogeneous endpoints for microorganisms, including functional as well as structural parameters, the use of sensitivity distributions is not considered to be applicable to this group of organisms, and a direct, expert evaluation of toxicity data was used instead. The soil concentration after sludge application was predicted for a number of scenarios and used as the predicted environmental concentration (PEC) in the risk characterization and calculation of risk quotients (RQ = PEC/PNEC). A LAS concentration of 4.6 mg/kg was used as the current best estimate of PNEC in all RQ calculations. Three levels of LAS contamination (530, 2,600, and 16,100 mg/kg), three half-lives (10, 25, and 40 d), and five different sludge loads (2, 4, 6, 8, and 10 t/ha) were included in the risk scenarios. In Denmark, the initial risk ratio would reach 1.5 in a realistic worst-case consideration. For countries not having similar sludge regulations, the estimated risk ratio may initially be considerably higher. However, even in the most extreme scenarios, the level of LAS is expected to be well beyond the estimated PNEC one year after application. The present risk assessment, therefore, concludes that LAS does not pose a significant risk to fauna, plants, and essential functions of agricultural soils as a result of normal sewage sludge amendment. However, risks have been identified in worst-case scenarios. PMID- 11491551 TI - Resource allocation-based life histories: a conceptual basis for studies of ecological toxicology. AB - Whereas ecological assessments of contaminants are concerned with populations and higher levels of organization, most mechanistic work in toxicology is directed at effects on individuals and their parts. We propose that studies based on individuals can be useful in ecological analysis of polluted systems when based on the concepts of resource allocation-based life history analysis. At the heart of the resource allocation approach is the concept of operative environments of individuals (i.e., environmental factors influencing birth, death, or migration). Contaminants can have strong influences on operative environments, modifying resource allocation strategies that reflect changes in energy assimilation and demands. By examining contaminant-induced responses of individuals from the perspective of changing operative environments, individual-based changes and population dynamics can be addressed in an ecologically rigorous manner. PMID- 11491552 TI - Effects of pre- and postnatal polychlorinated biphenyl exposure on metabolic rate and thyroid hormones of white-footed mice. AB - Energy budgets have proven to be a valuable tool for predicting life history from physiological data in terrestrial vertebrates, yet these concepts have not been applied to the physiological effects of contaminants. Contaminants might affect energy budgets by imposing an additional metabolic cost or by reducing the overall amount of energy taken in; either process will reduce the energy available for production (i.e., growth or reproduction). This study examined whole animal energetic effects of polychlorinated biphenyl (PCB) exposure in white-footed mice (Peromyscus leucopus). Exposure to PCBs is known to reduce concentrations of plasma thyroid hormones, and thyroid hormones exert strong control over the rate of energy metabolism in mammals. Peromyscus leucopus that were proven breeders were fed PCBs in their food at 0, 10, and 25 ppm. Through lactation, offspring were exposed to PCB from conception and were maintained on the maternal diet to adulthood. No effects were seen on energy metabolism (O2 consumption, measured in adulthood) or on growth, but there were large dose dependent decreases in thyroid hormone concentrations, particularly T4. The apparent disparity in our data between unchanged metabolic rates and 50% reductions in T4 concentrations can be rationalized by noting that free T3 (the fraction not bound to plasma protein) in treated mice was not significantly different from controls and that metabolism is most strongly influenced by free T3. Overall, this study did not demonstrate any energetic consequences of PCB exposure in P. leucopus at dietary concentrations up to 25 ppm. PMID- 11491553 TI - Effects of fungicides on thyroid function, metabolism, and thermoregulation in cotton rats. AB - Among the myriad of recent studies on endocrine-disrupting chemicals, relatively few involve thyroid disruption, and most of these address exposure/disruption during embryonic life. Of those involving adult vertebrates, the endpoints examined are thyroid measurements. Even though thyroid disruption could potentially interfere with energy metabolism and thermoregulation such that over winter survival might be compromised, the possible energetic consequences of these thyroid perturbations have not been investigated. We assessed thyroid function and measured resting metabolic rates of cotton rats chronically exposed to the fungicides vinclozolin or mancozeb. In addition, we measured norepinephrine-induced nonshivering thermogenesis and cold-induced thermogenesis and then cold-acclimated the mancozeb animals. Although thyroid hormone concentrations generally decreased, this was compensated for by an increase in thyroxine turnover (vinclozolin study only) such that thyroxine utilization rate was not different. In addition, there was no difference between the treated and control animals in any of the metabolic parameters measured. It is concluded that wild rodents exposed to these compounds are not energetically compromised. PMID- 11491554 TI - Bioavailability of lab-contaminated and native polycyclic aromatic hydrocarbons to the amphipod Corophium volutator relates to chemical desorption. AB - In the present study, the relationship between bioavailability of polycyclic aromatic hydrocarbons (PAHs) to benthic amphipods and the PAH desorption kinetics was examined. To that end, field-contaminated sediment was treated in three different ways. One subsample had no addition of PAHs and contained native PAHs only. To a second subsample, six PAHs (phenanthrene, fluoranthene, anthracene, pyrene, benzo[b]fluoranthene, and benzo[k]fluoranthene) were added in the laboratory. Two of the PAHs were added at higher concentrations to a third subsample, serving as a control for concentration-dependent uptake. Marine amphipods (Corophium volutator) were exposed to the three subsamples for a maximum of 25 d and were subsequently analyzed. Desorption kinetics were determined for both the lab-contaminated and the native PAHs. The biota-to sediment accumulation factor (BSAF) values of the individual native and lab contaminated PAHs correlated well with the rapidly desorbing fraction (R2 = 0.76). The BSAFs were 1.4 to 3.3 higher for the lab-contaminated PAHs compared with the native PAHs, while the difference between the rapidly desorbing fractions was a factor of 1.1 to 1.8. The BSAFs of the lab-contaminated PAHs in the second and third subsample were equal, indicating concentration-independent accumulation. The results suggest that lab-contaminated PAHs are more available to amphipods than native PAHs and that differences in bioavailability of lab contaminated and native PAHs to marine amphipods are related to differences in desorption behavior. PMID- 11491555 TI - Variability in acetylcholinesterase and glutathione S-transferase activities in Chironomus riparius Meigen deployed in situ at uncontaminated field sites. AB - An in situ system was used to expose fourth instar Chironomus riparius Meigen larvae for 48-h at 13 uncontaminated river sites across southeast England. Acetylcholinesterase (AChE) and glutathione S-transferase (GST) activities were measured in individual larvae recovered from these sites. Activities of both biomarkers varied almost twofold across the sites, with statistically significant differences detectable between sites. There were no clear relationships between biomarker activities and physical or chemical characteristics measured at each site. This study shows that biomarker measurements can be used successfully in conjunction with an in situ system. However, the results of statistically significant biomarker results must be treated with caution because natural variability in responses can occur even in the absence of toxicant exposure. PMID- 11491556 TI - Freeze concentration of ambient waters for toxicity testing. AB - We have developed a method to concentrate aqueous samples for toxicity testing. This method relies on the phenomenon of freezing exclusion, whereby solutes are rejected from the interstices of a growing ice crystal. Tenfold freeze concentration gave excellent recoveries of inorganic and organic analytes, phenol and ZnSO4 toxicity from spiked natural waters, and toxicity of both pre- and postdischarge municipal wastewater. Simultaneous 10-fold concentration of strong mineral or humic ambient matrices did not substantially modify the expressed toxicity of phenol or ZnSO4, and it did not seem to generate spurious toxicity to the marine bioassay organism used (Vibrio fischeri). Hundredfold freeze concentration permitted the quantification of low levels of ambient toxicity in a wide variety of natural waters using a rapid, inexpensive microbioassay. Precipitation of matrix elements may limit the degree of concentration that can be achieved with highly mineralized or strongly humic waters. This approach is well suited to ambient toxicity testing, because it is nonspecific and has low potential for solvent contamination. Furthermore, the low temperatures involved minimize volatilization and degradation of organic contaminants. PMID- 11491557 TI - Influences of aquatic plants on the fate of the pyrethroid insecticide lambda cyhalothrin in aquatic environments. AB - Aquatic exposure assessments for pesticides are generally based on laboratory studies performed in water alone or water sediment systems. Although aquatic macrophytes, which include a variety of bryophytes, macroalgae, and angiosperms, can be a significant component of many aquatic ecosystems, their impact on pesticide fate is generally not included in exposure assessments. To investigate the influence of aquatic plants on the fate and behavior of the pyrethroid insecticide lambda (lambda)-cyhalothrin, two laboratory experiments (to assess adsorption and degradation) and an indoor microcosm study (to assess fate under semirealistic conditions) were conducted. In the laboratory studies, adsorption to macrophytes was extensive and essentially irreversible, and degradation occurred rapidly by cleavage of the ester bond. In the indoor microcosm, which contained water, sediment, and macrophytes from a pond, degradation was also rapid, with DT50 and DT90 values of less than 3 and 19 h, respectively, for dissipation from the water column and of less than 3 and 56 h, respectively, for the whole system. For adsorptive and readily degraded pesticides like lambda cyhalothrin, we conclude that macrophytes have considerable influence on fate and behavior in surface waters. PMID- 11491558 TI - Heritability of resistance to individual contaminants and to contaminant mixtures in the sheepshead minnow (Cyprindon variegatus). AB - Resistance heritability (the additive genetic variance out of the total phenotypic variance, signifying a population's potential to genetically adapt to detrimental levels of contamination) was quantified in the sheepshead minnow (Cyprinodon variegatus). Heritability was estimated for tolerance to individual contaminants (phenanthrene, zinc) and to contaminant mixtures (phenanthrene plus zinc, and a complex mixture with three metals and three polycyclic aromatic hydrocarbons). Estimates were obtained from resemblances between relatives, both parent-offspring pairs, and families of sibs and half-sibs. Heritabilities determined from parent-offspring regressions averaged only 0.08 (scale, 0-1), whereas resemblance among full sibs yielded heritabilities averaging 0.85. The half-sib analysis yielded heritabilities of -0.01 (sire component) and 0.77 (dam component). This pattern in the magnitude of heritabilities indicates that heritabilities for the resistance of C. variegatus to these chemicals are low (with the high resemblances among sibs being due to common environmental and dominance genetic variation rather than additive genetic variation). The parent offspring regressions provide evidence that heritabilities may be lower if more contaminants are involved. Our results mean, then, that C. variegatus in contaminated environments is not likely to become resistant to these contaminants very rapidly, and that resistance may develop even more slowly as more contaminants become involved. PMID- 11491559 TI - Impact of sediment manipulation on the bioaccumulation of polycyclic aromatic hydrocarbons from field-contaminated and laboratory-dosed sediments by an oligochaete. AB - The accumulation kinetics of polycyclic aromatic hydrocarbons (PAHs) by the freshwater oligochaete Lumbriculus variegatus were measured for field contaminated and laboratory-dosed sediment. In addition, sediment manipulations typically used for homogenization and dosing in bioaccumulation assays were compared. Rather than an asymptotic approach to steady state, both resident and dosed PAH accumulation exhibited a peak during the 14-d assays, with steeper declines being noted for the lower-molecular-weight compounds. Lack of evidence of a peak for higher-molecular-weight PAHs may be due to slower kinetics and the short length of the assay. Relative to minimally mixed sediment, slurried sediment enhanced the accumulation of less-soluble resident PAHs, did not affect moderately soluble PAHs, and reduced the uptake of the more-soluble PAHs, fluorene and phenanthrene. Aging sediment after mixing reduced the availability of highly to moderately soluble resident PAHs but had no effect on less-soluble PAHs. A similar effect was noted for dosed PAHs, though a larger reduction in bioavailability was observed. Dosed PAH uptake clearance coefficients (ks) exceeded those of minimally mixed resident PAHs by factors of 3 to 4 for pyrene and 26 for benzo[a]pyrene. These results demonstrate that sediment manipulations and contamination history need to be considered when measuring PAH bioaccumulation. PMID- 11491560 TI - Toxicity of sediment-associated nitroaromatic and cyclonitramine compounds to benthic invertebrates. AB - The toxicity of nitroaromatic (2,4-diaminonitrotoluene [2,4-DANT] and 1,3,5 trinitrobenzene [TNB]) and 14C-labeled cyclonitramine compounds (hexahydro-1,3,5 trinitro-1,3,5-triazine [RDX] and octahydro-1,3,5,7-tetranitro-1,3,5,7 tetrazocine [HMX]) to the marine polychaete Neanthes arenaceodentata and the estuarine amphipod Leptocheirus plumulosus following 10- or 28-d exposures to spiked sediments was investigated. Organismal-level effects on survival, growth, and reproduction and cellular-level effects on apoptosis (programmed cell death) were evaluated. Because cyclonitramines have low affinity for sediment, overlying water was not exchanged in the RDX and HMX exposures. Nitroaromatics sorbed strongly to sediment, resulting in near complete resistance to solvent extraction. Cyclonitramines sorbed weakly to sediment, as more 14C-activity was found in the overlying water than in the sediment at exposure termination. No significant decrease in survival or growth was observed with cyclonitramines at initial sediment concentrations as high as 1,000 microg/g. Survival was significantly affected by nitroaromatics at nominal sediment concentrations as low as 200 microg/g, with L. plumulosus being more sensitive than N. arenaceodentata. Growth was significantly decreased at sublethal concentrations of 2,4-DANT for N. arenaceodentata. Reproduction, measured only with L. plumulosus, was significantly decreased only in the highest RDX treatment and also in the lower TNB treatment. However, no decrease was observed in higher concentrations of TNB. Body burden at exposure termination was below detection limit (1 microg/kg) for all compounds. Significant inhibition of apoptosis was not accompanied by significant decreases in growth or reproduction. Because of its critical function in many biological processes. alterations in this endpoint may result in adverse effects on the organism and could be used as an early indicator of toxicity. PMID- 11491561 TI - Life-cycle effects of sediment-associated 2,4,5-trichlorophenol on two groups of the midge Chironomus riparius with different exposure histories. AB - Effects of 2,4,5-trichlorophenol (TCP) on life-cycle traits of the midge Chironomus riparius and the ability of the midge to evolve tolerance to TCP were assessed using a reference group and a group preexposed to TCP during three generations, both originating from the same laboratory culture. F1 larvae of these groups were then exposed to nominal TCP concentrations of 51, 177, 355, and 532 micromol TCP/kg dry sediment and a control sediment in a life-cycle experiment. Most studied life-cycle traits (mortality, egg production, life span, male dry wt) were fairly insensitive to TCP, and significant effects were observed only at high (> or = 355 micromol/kg) concentrations. Larval development rate was variable, and in some cases it responded more readily to low TCP concentrations than other life-cycle parameters. Some of the observed responses were attributed to changes in food availability. No clear evidence of tolerance to 2,4,5-trichlorophenol was found, but the preexposed midges produced more eggs than the reference group. They also emerged earlier and at a smaller size than the reference midges. These differences between the midge groups suggest that some changes toward tolerance induction had occurred during the preexposure. PMID- 11491562 TI - Dietary uptake and elimination of selected polychlorinated biphenyl congeners and hexachlorobenzene in earthworms. AB - Earthworms (Eisenia foetida) were given food contaminated with hexachlorobenzene (HxCBz) and 20 tetra- to heptachlorinated biphenyls (PCBs). The PCBs were selected to cover a wide range of physicochemical properties. The uptake efficiencies, uptake rate constants, and elimination rate constants in worms varied between the 20 PCBs. The uptake efficiencies were between 10 and 68%. The PCBs that were eliminated fastest contained few chlorine atoms, had more polar characteristics, and were sterically hindered. Slowest was the elimination for less polar PCBs with six or seven chlorine atoms such as the non-ortho substituted PCB 169. Elimination half-lives varied between 6 and 25 d. The biomagnification factors (BMFs) for the PCBs and HxCBz averaged 0.1 based on lipids and 0.04 based on wet (worms) and dry (food) weight. Worm cocoons were analyzed for PCBs for the first time. The PCB pattern in the cocoons differed from the adult worms by the comparatively higher load of non-ortho PCBs. By studying congeners covering a large range of PCB physicochemical characteristics, their fate within the environmental and the factors influencing biological responses can be better understood. PMID- 11491563 TI - Cadmium uptake by earthworms as related to the availability in the soil and the intestine. AB - The free metal concentration in the soil solution is often considered a key parameter for metal uptake by and toxicity to soft-bodied soil organisms. The equilibrium partitioning theory, which assumes a relationship between the contaminant concentration in pore water and the contaminant concentration in the body tissue, can be used to describe uptake by earthworms. This theory has proved useful for organic chemicals, but its applicability is less clear for metals. In this study, the Cd concentration in soil pore water (pw) was varied by increasing the soil pH by the addition of lime (Ca(OH)2) and by adding manganese oxide (MnO2), which has a high metal binding capacity. Both lime (0.135% w/w) and MnO2 (1% w/w) decreased [Cd2+]pw by a factor of 25, while Cdworm was reduced only by a factor of 1.3 in lime-treated soils and 2.5 in MnO2-treated soils. Cadmium uptake was weakly related to the free metal concentration (R2adj = 0.66). Adding pH as an explanatory variable increased R2adj to 0.89, indicating that Cd uptake from pore water is pH dependent, which might be attributed to competition of protons and Cd at the surface of the earthwororm body. However, previous earthworm experiments in reconstituted groundwater showed a conspicuously smaller pH dependency of Cd uptake. The differences in metal uptake between earthworms in lime- and MnO2-treated soils are therefore more likely to reflect the predominance of pH-independent intestinal uptake of Cd. Equilibrating the soil with a solution of 0.01 M CaCl2 and 0.1 M triethanolamine (buffered at pH 7.2), simulating the conditions prevailing in the worm intestine, yielded free Cd concentrations that were closely (R2adj = 0.83) and linearly related to the Cd concentration in the earthworm tissue. PMID- 11491564 TI - Abnormalities in sexual development of the amphipod Gammarus pulex (L.) found below sewage treatment works. AB - Increasing numbers of widely used industrial, agricultural, and natural chemicals are known to elicit endocrine-disrupting effects in a wide range of vertebrate and invertebrate species. The objective of this study was to determine whether the sexual development of the freshwater crustacean Gammarus pulex (L.) was affected below sewage treatment works (STW) previously known to contain endocrine disrupting chemicals in their effluent. The gonadal structure, external sexual characteristics, and size of gammarids from exposed sites were compared to those of gammarids from a reference site. No significant difference was found in the gonadal structure of males collected below two STW. However, a highly significant number of females collected from a site known to elicit high estrogenic responses in vertebrates displayed an abnormal structure of oocytes in vitellogenesis. Body size was significantly shorter and male/female size differential was significantly reduced below one of the STW. Analysis of gnathopod and genital papillae length data suggests that different allometric relationships of these organs to body size exist between sample sites. PMID- 11491565 TI - An assessment of the toxicity of phthalate esters to freshwater benthos. 1. Aqueous exposures. AB - Tests were performed with the freshwater invertebrates Hyalella azteca, Chironomus tentans, and Lumbriculus variegatus to determine the acute toxicity of six phthalate esters, including dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), butylbenzyl phthalate (BBP), di-n-hexyl phthalate (DHP), and di-2-ethylhexyl phthalate (DEHP). It was possible to derive 10-d LC50 (lethal concentration for 50% of the population) values only for the four lower molecular weight esters (DMP, DEP, DBP, and BBP), for which toxicity increased with increasing octanol-water partition coefficient (Kow) and decreasing water solubility. The LC50 values for DMP, DEP, DBP, and BBP were 28.1, 4.21, 0.63, and 0.46 mg/L for H. azteca; 68.2, 31.0, 2.64, and > 1.76 mg/L for C. tentans; and 246, 102, 2.48, and 1.23 mg/L for L. variegatus, respectively. No significant survival reductions were observed when the three species were exposed to either DHP or DEHP at concentrations approximating their water solubilities. PMID- 11491566 TI - An assessment of the toxicity of phthalate esters to freshwater benthos. 2. Sediment exposures. AB - Seven phthalate esters were evaluated for their 10-d toxicity to the freshwater invertebrates Hyalella azteca and Chironomus tentans in sediment. The esters were diethyl phthalate (DEP), di-n-butyl phthalate (DBP), di-n-hexyl phthalate (DHP), di-(2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP), diisodecyl phthalate (DIDP), and a commercial mixture of C7, C9, and C11 isophthalate esters (711P). All seven esters were tested in a sediment containing 4.80% total organic carbon (TOC), and DBP alone was tested in two additional sediments with 2.45 and 14.1% TOC. Sediment spiking concentrations for DEP and DBP were based on LC50 (lethal concentration for 50% of the population) values from water-only toxicity tests, sediment organic carbon concentration, and equilibrium partitioning (EqP) theory. The five higher molecular weight phthalate esters (DHP, DEHP, DINP, DIDP, 711P), two of which were tested and found to be nontoxic in water-only tests (i.e., DHP and DEHP), were tested at single concentrations between 2,100 and 3,200 mg/kg dry weight. Preliminary spiking studies were performed to assess phthalate ester stability under test conditions. The five higher molecular weight phthalate esters in sediment had no effect on survival or growth of either C. tentans or H. azteca, consistent with predictions based on water-only tests and EqP theory. The 10-d LC50 values for DBP and H. azteca were >17,400, >29,500, and >71,900 mg/kg dry weight for the low, medium, and high TOC sediments, respectively. These values are more than 30x greater than predicted by EqP theory and may reflect the fact that H. azteca is an epibenthic species and not an obligative burrower. The 10-d LC50 values for DBP and C. tentans were 826, 1,664, and 4.730 mg/kg dry weight for the low, medium, and high TOC sediments, respectively. These values are within a factor of two of the values predicted by EqP theory. Pore-water 10-d LC50 values for DBP (dissolved fraction) and C. tentans in the three sediments were 0.65, 0.89, and 0.66 of the water-only LC50 value of 2.64 mg/L, thereby agreeing with EqP theory predictions to within a factor of 1.5. The LC50 value for DEP and C. tentans was >3,100 mg/kg dry weight, which is approximately 10x that predicted by EqP theory. It is postulated that test chemical loss and reduced organism exposure to pore water may have accounted for the observed discrepancies with EqP calculations for DEP PMID- 11491567 TI - Kinetics, dose-response, excretion, and toxicity of methylmercury in free-living Cory's shearwater chicks. AB - Methylmercury (MHg) kinetics, dose-response, excretion, and toxicity were experimentally evaluated and compared between small (one to two weeks old) and large (six to seven weeks old), free-living Cory's shearwater chicks. The half time for the terminal elimination phase of MHg in blood (5.7 d) and the average percentage of ingested MHg deposited in the blood volume (12%) were independent of the age at exposure. Therefore, these data were employed to derive a relationship between steady-state blood concentrations and dietary intake of MHg in bird chicks. Plumage:blood ratios were independent of dose and could be used as partition coefficients. Dose-response relationships in plumage and blood were linear over the wide range of exposures employed. Blood dose-responses of MHg in small and large chicks were similar. Excretion percentages into the final plumage varied between 42 and 60% of intake. The body condition of experimental chicks did not indicate sublethal toxicity of the doses administered; hence, the exposure levels provide maximum avian no-observed-adverse-effect levels for external symptoms in wild seabird chicks. PMID- 11491568 TI - Toxicity evaluation of natural and synthetic phenanthrenes in aquatic systems. AB - Seven natural 9,10-dihydrophenanthrenes were isolated from the common reed Juncus effusus by means of chromatographic processes and identified by spectroscopic means. Furthermore, mimics of natural isolated compounds were synthesized to try to evaluate the influence of functional groups on the dihydrophenanthrene skeleton. Syntheses of compounds were based on the cross-coupling of 1-(2-iodo-5 methoxy)phenyl-ethanol with variously substituted iodobenzenes by zerovalent nickel. All the chemicals were tested to evaluate their effects on freshwater organisms from different trophic levels. Toxicity tests were performed on reducers (the bacterium Escherichia coli); producers (the alga Raphidocelis subcapitata, previously known as Selenastrum capricornutum); and consumers including a rotifer (Brachionus calyciflorus), a cladoceran (Daphnia pulex), and an anostracan (Thamnocephalus platyurus). Results suggested no one organism was uniquely sensitive to the chemicals tested. Toxicity depended on the kind and position of substituents on the aromatic skeleton. PMID- 11491569 TI - Heavy metals in soft tissue of white-tailed eagles found dead or moribund in Germany and Austria from 1993 to 2000. AB - Residues of the potentially toxic metals lead, mercury, and cadmium were analyzed in liver and kidney tissue of 61 free-ranging white-tailed eagles (Haliaeetus albicilla) found dead or moribund in Germany and Austria between 1993 and 2000. Highest values and the widest range were detected for lead in liver and for mercury in kidney tissue. Lead concentrations considered to induce lethal lead poisoning (>5 ppm wet wt) were determined in 28% of liver samples. Lead fragments were detected it the gizzards of two specimens, presumably having died from lead intoxication. Histopathological findings in a recently dead white-tailed eagle indicating acute lead exposure comprise degenerative Purkinje cells in the cerebrellum and inclusion bodies in renal tubular cells. Mercury residues in organs are decreasing compared to former studies in periods when organomercury compounds were used as seed dressing. All cadmium values were low or are at background levels in white-tailed eagles. The present study clearly identities lead as a toxic metal poison in white-tailed eagles in Germany and Austria. PMID- 11491570 TI - Effects of three vertebrate hormones on the growth, development, and reproduction of the tomato moth, Lacanobia oleracea L. (Lepidoptera: Noctuidae). AB - In recent years, concern has been growing that numerous manmade chemicals entering the environment are capable of mimicking endogenous hormones in wildlife. In an attempt to define and evaluate the possible impact of endocrine disrupting substances (EDS) on insects, three vertebrate hormones were tested for their effects on growth, development, and reproduction of the tomato moth. Lacanobia oleracea. Dietary administration of estrogen or thyroxine caused a significant increase in the length of the third, fourth, fifth, and sixth larval stadia (p < 0.001). The mean time for development of the pupa, however, was not significantly different between treatments. Relative to the control groups, a significant (p < 0.05) reduction in the mean weights of fifth- and sixth-instar larvae was also observed when larvae were exposed to estrogen or thyroxine and in pupae derived from insects exposed to thyroxine or testosterone (p < 0.001). Despite this, the number of larvae that survived to adulthood was not affected by any of the treatments; neither was the pupal sex ratio affected. However, exposure of larvae to testosterone significantly (p < 0.05) increased the number of deformed pupae. In addition, the reproductive potential of adults derived from the testosterone treatment was markedly reduced. Exposure of L. oleracea larvae to this steroid caused a highly significant (p < 0.001) decrease in egg production. coupled with a significant (p < 0.05) decrease in egg viability. The physiological effects observed in L. oleracea and their possible causes are presented in this paper, and the likely impact of EDS and their effects on terrestrial invertebrates are discussed. PMID- 11491571 TI - Assessing acute and chronic copper risks to freshwater aquatic life using species sensitivity distributions for different taxonomic groups. AB - Using copper as an example, we present a method for assessing chemical risks to an aquatic community using species sensitivity distributions (SSDs) for different taxonomic groups. This method fits probability models to chemical exposure and effects data to estimate the percentage of aquatic species potentially at risk and expands on existing probabilistic risk assessment methodologies. Due to a paucity of chronic toxicity data for many chemicals, this methodology typically uses an acute-chronic ratio (ACR) to estimate the chronic effects distribution from the acute effects distribution. We expanded on existing methods in two ways. First, copper SSDs were developed for different organism groups (e.g., insects, fish) that share similar sensitivities or ecological functions. Integration of exposure and effects distributions provides an estimate of which organism groups may be at risk. These results were then compared with a site-specific food web, allowing an estimation of whether key food web components are potentially at risk and whether the overall aquatic community may be at risk from the perspective of ecosystem function. Second, chronic SSDs were estimated using the relationship between copper ACRs and acute toxicity (i.e., the less acutely sensitive a species, the larger the ACR). This correction in the ACR removes concerns previously identified with use of the ACR and allows evaluation of a significantly expanded chronic data set with the same approach as that for assessing acute risks. PMID- 11491572 TI - Model selection and parameterization of the concentration-response functions for population-level effects. AB - As concentration response functions for chronic population-level effects of pollutant chemicals, three mathematical models were presented and examined for goodness of fit to published toxicological data that estimated the population level effects of chemicals in terms of the intrinsic rate of population growth (r). Among the examined concentration-r functions, the power function model, that is, r(x) = r(0)[1 - (x/alpha)beta], in which x is the exposure concentration and alpha and beta are parameters, performed with the best fit to each data set. The power function model is characterized by two parameters representing the absolute value of toxicity, alpha, and the curvature of responses, beta. The bootstrap simulation, conducted on the entire data set consisting of all published data that we collected, indicated that the observed variance of beta among actual data sets could be mostly explained by the random error variation generated from the bootstrap resamplings. The generic beta value, determined from the entire data set and expected to denote the best estimate of beta if the variability of beta was completely due to random sampling error, was estimated as 1.84. It was implied that the response of the intrinsic rate of natural increase (r) to chemical exposure was nearly quadratic in many cases. PMID- 11491573 TI - Radiation induced peroxidative damage: mechanism and significance. AB - An interest has been generated in free radicals after the discovery of superoxide dismutase. These free radicals cause a number of diseases and are involved in the detrimental effect of ionizing radiation. Efforts have been made to understand their role in damage and death of the cell using lipid peroxidation process. Lipid peroxidation is an important effect of radiation on membranes, which apart from DNA, are critical targets of radiation action. This paper addresses the basic mechanism of radiation induced lipid peroxidation. Various factors, which determine the mode and magnitude of lipid peroxidation, are also discussed. Lipid peroxidation is shown to have importance in understanding the modifications of radiation effects. Efforts are made to show similarities between radiolytic and non-radiolytic lipid peroxidation. Recent findings related to the close link between radiation-induced lipid peroxidation and apoptosis are likely to open new avenues for future research and to develop new approaches for radiomodification of biological effects. PMID- 11491574 TI - Role of garlic (Allium sativum L.) in human and plant diseases. AB - A resurgence of interest in garlic due to recent revelations of its beneficial effects in the treatment of various human and plant diseases and also due to validation of claims made in traditional systems of medicine has resulted a plethora of publications on different aspects of garlic in recent years. Chemical constituents of garlic and their variations on the methods of isolation have been discussed in the present review. Effect of garlic and its constituents against various human and plant pathogenic and saprophytic microorganisms has also been reviewed. PMID- 11491575 TI - Comparative functional characterization of mouse bone marrow-derived mast cells and peritoneal mast cells in response to non-immunological stimuli. AB - The cultured mouse mast cells that are dependent on spleen-derived factor for their proliferation and maintenance and have been shown to be similar to mucosal mast cells in terms of their T-cell dependence and histochemical staining characteristics. Mast cell heterogeneity has been confirmed by functional characterization of mouse bone marrow-derived mast cells (MBMMC) and mouse peritoneal mast cells (MPMCs). MPMCs released around 30% of histamine when stimulated with compound 48/80 whereas MBMMC were almost unresponsive to the same stimulus. Calcium Ionophore A23187 on the other hand, released histamine in dose dependent manner from MBMMC. The study was undertaken to investigate the effect of antiallergic drug, disodium cromoglycate (DSCG), a synthetic cromone and quercetin, a plant-derived flavonoid on Ca ionophore A23187 induced histamine release from MBMMC. MBMMCs were almost unresponsive to DSCG whereas Ca Ionophore induced histamine release was blocked by Quercetin. The results indicate that response of mast cells at one anatomic site to a given stimulus does not necessarily predict the response of mast cells at a different anatomic location to the same stimulus. It shows functional heterogeneity within a single species. So, it cannot be assumed that antiallergic compounds stabilizing mast cells in one tissue site or organ will be equally efficacious against mast cells in other sites. PMID- 11491576 TI - Influence of influenza viral infection on airway smooth muscle activity. AB - The contractility of airway smooth muscle (ASM) plays an important role in pathophysiology of several bronchial disorders. Increased contraction of ASM during asthma and respiratory viral infection has been attributed to the release of mediators acting through different receptors. In the present study, influence of influenza type A virus (H1N1) infection has been examined on ASM responsiveness to various bronchoactive agents e.g. adenosine, histamine, 5 hydroxytryptamine (5-HT) and isoproterenol in an organ bath set up for isolated tissue preparation. The contractile effect of adenosine, histamine and 5-HT was enhanced, however, relaxant response of isoproterenol was attenuated with the duration following viral exposure. The most prominent response was observed 48 to 72 hr after infection and tissues from multiple exposure to virus infected animals showed the maximum contractile response. Results demonstrated the deleterious effect of viral infection on ASM function and the findings will be helpful in understanding the mechanism of influenza virus induced bronchoconstriction. PMID- 11491577 TI - Neurochemical studies on Indian Hypericum perforatum L. AB - The effect of acute administration of 50% standardised ethanolic extract of Indian Hypericum perforatum (IHp) was studied on the rat brain concentrations of monoamines and their metabolites in five different brain regions, viz. hypothalamus, hippocampus, striatum, pons-medulla and frontal cortex by a HPLC technique. IHp extract was administered at the doses of 50 and 200 mg/kg, p.o. and the brain monoamines were assayed after 30 min of the treatment. IHp treatment significantly decreased the levels of serotonin (5-HT) and its metabolite 5-hydroxy indole acetic acid (5-HIAA) and 5-HT turnover in all the brain regions assayed. On the other hand, IHp treatment significantly augmented the levels of norepinephrine (NE) and its metabolite methylhydroxy phenyl glycol (MHPG) and NE turnover in all the brain regions studied. Similarly, the levels of dopamine (DA) were also significantly augmented in the hypothalamus, striatum and frontal cortex. Likewise, the levels of dihydroxyphenyl acetic acid (DOPAC), the major metabolite of DA, were also increased in these brain areas. Pharmacological studies with IHp extract have shown two major behavioural actions, namely, anxiolytic and antidepressant effects. The present findings tend to rationalise these observations, reduced 5-HT activity being consonant with anxiolytic and increased NA and DA activity being consonant with antidepressant action. PMID- 11491578 TI - Anti-inflammatory and analgesic activity of Indian Hypericum perforatum L. AB - A standardised 50% aqueous ethanolic extract of the Indian variety of Hypericum perforatum (IHp) was examined for its putative anti-inflammatory and analgesic activity at the doses of 100 and 200 mg/kg, po. The experimental paradigms used were carrageenan induced pedal edema and cotton pellet induced granuloma for anti inflammatory activity, whereas the tail flick, hot plate and acetic acid induced writhing methods were used to asses analgesic activity. Indomethacin (20 mg/kg, ip) was used as the standard anti-inflammatory drug. Pentazocine (10 mg/kg, ip) and aspirin (25 mg/kg, ip), both clinically used analgesics, were used as standard analgesics for comparison. IHp extract showed significant anti inflammatory and analgesic activity at both dose levels, in all the paradigms used. Additionally, IHp potentiated the anti-inflammatory activity of indomethacin and analgesic activities of pentazocine and aspirin. PMID- 11491580 TI - Effect of Convolvulus pluricaulis Chois on gastric ulceration and secretion in rats. AB - Convolvulus pluricaulis is an indigenous plant commonly mentioned in Ayurveda, an ancient system of Indian medicine, as a rasayana which is mainly advocated for use in rejuvenation therapy. The present study was conducted to evaluate the potential anti-ulcerogenic effect of juice of fresh whole plants of C. pluricaulis (CPJ) against various experimental gastric ulcer models induced by ethanol, aspirin, 2 hr cold restraint stress and 4 hr pyloric ligation in rats. The drug was given orally twice daily for five days in the doses of 375 and 750 mg/kg body weight. CPJ showed anti-ulcerogenic effect at both doses in all the experimental gastric ulcer models and was comparable to the reference drug sucralfate (250 mg/kg). Gastric juice secretion and mucosal studies were undertaken to find out the possible mechanism of action of antiulcer effect by studying its effects both on offensive and defensive mucosal factors. The antiulcerogenic effect of CPJ was found to be due to augmentation of mucosal defensive factors like mucin secretion, lifespan of mucosal cells and glycoprotiens rather than on the offensive factors like acid-pepsin. PMID- 11491579 TI - Anti-stress activity of Indian Hypericum perforatum L. AB - Indian Hypericum perforatum (IHp) was investigated on a 14-day mild, unpredictable and inescapable foot shock stress (FSS) induced perturbations in behaviour (depression), suppressed male sexual behaviour and cognitive dysfunction in albino rats. Gastric ulceration, and adrenal gland and spleen weights, were also used as the stress indices. Panax ginseng (PG) was used as the standard adaptogenic agent for comparison. FSS induced marked gastric ulceration, significant increase in adrenal gland weight with concomitant decrease in spleen weight. Chronic stress also suppressed male sexual behaviour, induced behavioural depression (Porsolt's swim despair test and learned helplessness test) and cognitive dysfunction (attenuated retention of learning in active and passive avoidance tests). All these FSS induced perturbations were attenuated dose dependently by IHp (100 and 200 mg/kg, po) and PG (100 mg/kg, po). The results indicate that IHp has significant anti-stress activity, qualitatively comparable to PG, against a variety of behavioural and physiological perturbations induced by chronic stress, which has been proposed to be a better indicator of clinical stress than acute stress, and may indicate adaptogenic activity. PMID- 11491581 TI - Effect of Maharishi Amrit Kalash an ayurvedic herbal mixture on lipid peroxidation and neuronal lipofuscin accumulation in ageing guinea pig brain. AB - The effects of ayurvedic herbal mixture Maharishi Amrit Kalash(MAK) were studied on brain lipid peroxidation, oxygen consumption, and lipofuscin accumulation in 10 months and 32 months old guinea pigs. Brain regions studied were cerebral cortex, hypothalamus, cerebellum and spinal cord. Parameters assessed were lipid peroxidation, oxygen consumption, and lipofuscin accumulation. The endogenous lipid peroxide was found to be increased significantly (P < 0.05) in the 32-month old animals. Neuronal lipofuscin accumulation in the neurons of cerebral motor cortex, cerebellum and cervical spinal cord was increased (P < 0.05) in the older animals. Oxygen consumption was found to be decreased significantly(P < 0.05) in the 32-month old guinea pigs. Treatment with MAK at a dose of 500 mg/kg body weight daily for two months reduced the lipid peroxidation and lipofuscin pigment accumulation significantly in brain regions and it also helped in restoring the normal oxygen consumption in the older animals. This indicates antioxidant properties of MAK. PMID- 11491582 TI - N-Phthaloyl gamma-aminobutyric acid affects biochemical circadian rhythms in Wistar rats. AB - N-Phthaloyl gamma-aminobutyric acid (P-GABA) was administered to Wistar rats and 24 hr rhythms of glucose, cholesterol, total protein and lactic acid levels in blood were studied under semi-natural light dark conditions. P-GABA administration caused desynchronisation of the rhythms; while glucose and lactic acid rhythms were advanced, cholesterol and total protein rhythms were delayed. Since GABA is being involved in conveying dark information to the clock, exogenous administration of P-GABA may reduce the photic information received by the clock. The results could be explained by slightly less than 1 hr daily delays (or) advances respectively which would bring the peak times to the points 21 days after the start of administration. PMID- 11491583 TI - Blood anticoagulant sulphated polysaccharides of the marine green algae Codium dwarkense (Boergs.) and C. tomentosum (Huds.) Stackh. AB - Cold water extracts of marine green algae Codium dwarkense and C. tomentosum were precipitated with different molar concentrations of KCl and were subjected to anion exchange and gel filtration chromatography. Both the species yielded sulphated arabinan through bioassay-guided purification and both were chemically identified as a polymer of alpha-L-arabinofuranose. Products were assayed for their blood anticoagulant activity using PT, APFT and TT tests and found that they differed in the potency of activity though they are chemically identical. Bioassay-guided purification of cold water extract of C. tomentosum yielded sulphated arabinan and sulphated arabinogalactan. PMID- 11491584 TI - Ultrastructural studies on nodules induced by pyrimidine auxotrophs of Sinorhizobium meliloti. AB - Twenty three pyrimidine auxotrophs of Sinorhizobium meliloti Rmd201 were generated by random mutagenesis with transposon Tn5. On the basis of biochemical characters these auxotrophic mutants were classified into car, pyrC and pyrE/pyrF categories. All auxotrophs induced white nodules which were ineffective in nitrogen fixation. Light and electron microscopic studies revealed that the nodules induced by pyrC mutants were more developed than the nodules of car mutants. Similarly the nodules induced by pyrE/pyrF mutants had more advanced structural features than the nodules of pyrC mutants. The nodule development in case of pyrE/pyrF mutants was not to the extent observed in the parental strain. These results indicated that some of the intermediates and/or enzymes of pyrimidine biosynthetic pathway of S. meliloti play a key role in bacteroidal transformation and nodule development. PMID- 11491585 TI - Effect of anxiolytics on blood sugar level in rabbits. AB - Anxiety disorders are more prevalent not only in normal individuals but also in diabetes mellitus. Diazepam, a benzodiazepine, and buspirone, an azaspirodecanedione, are the most often prescribed anxiolytics. Present study was aimed to investigate the effect of diazepam and buspirone on the blood sugar levels in rabbits. Buspirone (0.5 mg/kg/day p.o.) and diazepam (0.6 mg/kg/day p.o.) did not affect the glucose levels in rabbits even after one month of treatment. Present findings suggest that these two anxiolytics have minimal effect on blood sugar control. PMID- 11491586 TI - Haematinic effect of Hygrophila spinosa T. Anderson on experimental rodents. AB - Ethanolic extract of the aerial parts of H. spinosa a semiwoody herb was examined on male albino rats for certain haematological changes. The extract (100 & 200 mg/kg, po) significantly increased the haemoglobin, haematocrit, RBC and total WBC, as compared with vehicle treated control rat haemogram. In anemic male albino rats, the extract significantly increased haemoglobin, haematocrit and RBC count. Serum iron and serum total iron binding capacity were significantly decreased in H. spinosa extract treated anemic rats as compared with those in the vehicle treated anemic control rats. These findings demonstrated the haematinic effect of H. spinosa extract on experimental animals. PMID- 11491587 TI - Choice of oviposition site between surface of the medium and paper in four Indian species of Drosophila. AB - Choice of oviposition site between surface of the medium and paper positioned vertically on the medium by females was studied in four species of Drosophila: D. ananassae, D. bipectinata, D. malerkotliana, and D. biarmipes. Several geographic strains of these species were used. Females of all the four species laid greater proportions of eggs on the surface of medium as compared to surface of the paper. Thus all the four species tested show preference for oviposition on surface of medium. When the comparison is made for oviposition on paper among the four species, D. ananassae lays more eggs on the surface of paper as compared to other three species. PMID- 11491588 TI - Propagation of Coriandrum sativum L. through somatic embryogenesis. AB - A highly embryogenic callus was obtained from hypocotyl segments of Coriandrum sativum L. when cultured in the medium consisting of MS + H vitamins (MSH). Induction of somatic embryos required 2,4-dichlorophenoxyacetic acid or napthalene acetic acid. Germination of fully developed embryos was accomplished by subculture on half strength MSH medium containing benzylamino purine 0.05 mg/L. Plantlets developed from somatic embryos were transferred to soil and were successfully flowered. PMID- 11491591 TI - Near drowning in cold water. AB - Drowning and near drowning is a common cause of accidental death all over the world; specially in road traffic accidents over bridges, swimming pool and boat tragedies. Cold water drowning resulting in hypothermia can lead to instant death before actual drowning. Five cases of near drowning (ND) in cold water, who presented with varied clinical picture like coma with decerebrate rigidity and fixed dilated pupils, hypertension with coma and delayed pulmonary oedema (Secondary drowning) are reported. Energetic management with continuous positive airway pressure was very rewarding in all patients with ND except in one who had transient organic psychosis persisting for two weeks followed by minimal cognitive defect in the form of slow mentation, lack of drive and mild irritability (Bender Gestald Test Score of 53). We have tried to analyse some of the clinical features of ND and the sequel associated with it. The management as well as recent developments in the field are also discussed. PMID- 11491619 TI - Severe hyponatremia in a chronic schizophrenic patient. PMID- 11491622 TI - The biologic approach to restorative dentistry. PMID- 11491623 TI - Pulp-dentin biology in restorative dentistry. Part 1: normal structure and physiology. AB - Considerable knowledge has accumulated over the years on the structure and function of the dental pulp and dentin. Some of this knowledge has important clinical implications. This review, which is the first of seven articles, will be limited to those parts of the normal structure and physiology of the pulp and dentin that have been shown to result in, or are likely lead to, tissue reactions associated with the clinical treatment of these tissues. Although certain normal structures will be highlighted in some detail, a basic knowledge of pulpal and dentinal development and structure is a prerequisite for an understanding of this text. PMID- 11491624 TI - Calcific metamorphosis: a challenge in endodontic diagnosis and treatment. AB - Calcific metamorphosis (CM) is seen commonly in the dental pulp after traumatic tooth injuries and is recognized clinically as early as 3 months after injury. Calcific metamorphosis is characterized by deposition of hard tissue within the root canal space and yellow discoloration of the clinical crown. Opinion differs among practitioners as to whether to treat these cases upon early detection of CM or to observe them until symptoms or radiographic signs of pulpal necrosis are detected. In this review, the clinical, radiographic, and histopathologic appearance of CM is described; a review of the literature is presented to address these issues in an attempt to establish a sound rationale for treatment. Approximately 3.8% to 24% of traumatized teeth develop varying degrees of CM. Studies indicate that of these, approximately 1% to 16% will develop pulpal necrosis. Most of the literature does not support endodontic intervention unless periradicular pathosis is detected or the involved tooth becomes symptomatic. It may be advisable to manage cases demonstrating CM through observation and periodic examination. PMID- 11491625 TI - Wettability of denture materials. AB - OBJECTIVE: To promote denture retention and denture comfort, denture materials should possess adequate wettability. This in vitro study investigated the wettability of nine commercially available dental materials. METHOD AND MATERIALS: Four denture base materials, two denture hard lining materials, and three denture soft lining materials (with and without varnish treatment) were tested. The wettability measurements were made using the dynamic contact angle analysis technique. The equilibrium and hysteresis angles obtained were used for the comparisons. RESULTS: The equilibrium contact angle (thetae) ranged from 63.9 (Permaflex + varnish) to 81.0 degrees (Mollosil + varnish). The differences observed among the materials tested were statistically significant. The contact angle hysteresis ranged from 16.0 (SR 3/60 Triplex) to 51.2 degrees (Mollosil), and there was a statistically significant difference among the materials. CONCLUSION: The heat-polymerized soft lining materials exhibited the greatest equilibrium contact angle, the autopolymerized soft liner had the lowest value, and the denture base materials had intermediate values. The soft liners showed greater contact angle hysteresis than all other materials. The use of varnish altered the wetting characteristics. PMID- 11491626 TI - Clinical evaluation of glass-ionomer/resin-based hybrid materials used as pit and fissure sealants. AB - OBJECTIVE: The objective of the present study was to clinically evaluate the hybrid materials Vitremer and Dyract when used as pit and fissure sealants. METHOD AND MATERIALS: The materials were applied in pairs on the mandibular permanent first molars of 100 children, 7 to 8 years old. RESULTS: After 12 months of sealing, the clinical evaluation presented rates of 95.9% and 85.7% of complete retention for Dyract and Vitremer, respectively, indicating a statistically significant difference. The hybrid materials presented a statistically significant protective effect against caries compared with the control group at 6 and 12 months. CONCLUSION: The hybrid materials were able to control occlusal caries. PMID- 11491627 TI - Oral rehabilitation of a bulimic patient: a case report. AB - Bulimia nervosa is among the most common health problems in contemporary society. It is a self-induced weight loss syndrome associated with distinct dental manifestations involving physical and psychologic symptoms. It is characterized by recurrent binge-purge episodes that occur at least once a day. This article describes the complete-mouth rehabilitation of a bulimic patient with a generalized enamel erosion of her dentition and a poor esthetic appearance. Porcelain-fused-to-metal restorations were used as the definitive treatment. Good esthetics and high self-esteem were the final results. Comprehensive restorative therapy was applied in this clinical case report to achieve both function and esthetics in a demanding situation. PMID- 11491628 TI - Tongue piercing: a restorative perspective. AB - The implication of a traumatic injury to a permanent molar tooth as a complication by tongue piercing with ornamental jewelry is reported. An appropriate restorative management of the fractured tooth is described. The dentist's role in prevention and treatment of unfavorable complications is outlined. The rationales for selection of specific treatment modalities, in view of the degree of damage to hard dental tissues, are discussed. The range of hard tissue injuries resulting from tongue jewelry varies from a simple crack propagating into the enamel to a fractured tooth. A variety of factors must be considered before employing the available restorative methods. PMID- 11491629 TI - The good practitioner: the story of Brent L. Benkelman. AB - Dr Brent Benkelman has practiced oral surgery in Manhattan, Kansas, since 1971. Having grown up in a tiny farming town in western Kansas, he opted for the lifestyle of a smaller community after graduating from the University of Missouri at Kansas City School of Dentistry in 1966 and completing his training in oral and maxillofacial surgery in 1969. His experiences since then have revolved around family, practice, and community. With his family as his first priority, he has participated in various community activities, including a church-operated food pantry, an emergency shelter for the homeless, and Habitat for Humanity. He was nominated as a moral exemplar in dentistry for his dedicated commitment to his patients and because he understands and cares about the complexities of his patients as human beings, placing his own financial gain secondary to the interests of his patients. PMID- 11491630 TI - Lymphangioma (lymphatic malformation). PMID- 11491631 TI - Wet and dry deep cavity preparations compared by a novel odontoblast culture technique. AB - The human odontoblast's unique cellular extension within dentin does not easily allow culturing by traditional methods. This study leaves these cells in their natural position in the dentin. Deep preparations were made through the occlusal surfaces of extracted human third molars. The crowns were separated from the roots and the pulps gently teased from the chambers, leaving the odontoblast layer intact. These inverted pulp chambers were then incubated in cell culture medium for 2 to 4 days. Trypan blue staining was used to detect non-vital odontoblasts, and the differentiation between vital and nonvital cells was verified by SEM and toluidine vital staining. Control teeth and areas not adjacent to the preparation showed no blue staining, indicating intact cells. Areas of nonvital cells were greatest with wider preparation. Irrigation decreased odontoblast death with wide preparations. No difference due to irrigation was detected in narrow preps. A comparison of wet preparations to which heat was applied versus dry preparations showed statistically similar results. This study provides a simple in vitro method for the study of odontoblasts with their processes intact within dentin. PMID- 11491632 TI - Identification of GM3 as a marker of therapy-resistant periradicular lesions. AB - The purpose of this study was to analyze the profile of glycosphingolipids (GSLs) in periradicular lesions refractory to endodontic treatment. Sixteen periapical lesions were removed surgically from patients (experimental group) and compared with 10 samples of periodontal ligament removed from extracted intact third molars (control group). After the GSLs extraction and purification procedures were performed the neutral and acidic GSL fractions were analyzed by high performance thin-layer chromatography and quantified by densitometry. Data reported herein show that: (i) tissues in the experimental group presented about twice as much GSLs as the control group; (ii) lesion tissues express lactoneotetraosylceramide, and lactofucopentaosyl (IV) ceramide, whereas these neutral GSLs are absent in normal tissues; and (iii) normal tissues express GT1b, whereas lesions cells do not express this ganglioside. In contrast lesion tissues express GM3, which is conspicuously absent in normal tissues. PMID- 11491633 TI - Enamel matrix derivative prolongs primary osteoblast growth. AB - Enamel matrix derivative (EMD) secreted by cells of the epithelial root sheath plays an important role in cementogenesis and periodontal tissue formation. The mechanisms by which EMD influences cell function are not known. The purpose of this study was to determine the effect of EMD on cell growth of primary mouse osteoblasts. Osteoblasts were digested from 6- to 8-day-old mouse calvaria and plated into 6-well cell culture plates at an initial density of 5000 cells/cm2. After 24-h incubation with Dulbecco's modified eagle medium (DMEM) supplemented with 10% fetal bovine serum, cells were incubated in three different groups of media: DMEM only as control, DMEM with 25 microg/ml EMD, and DMEM with 100 microg/ml EMD. At days 3, 7, 10, and 14, the total cell number per well was calculated, and cell morphology was examined. At each observation period the number of cells in the EMD groups was significantly greater (ANOVA, p < 0.01) than that in the control group. EMD had a greater effect on osteoblast survivor in the higher concentration than in the lower concentration. Furthermore normal morphology of the primary osteoblasts was maintained in the EMD groups. These results suggest that EMD prolongs primary osteoblast growth and may have an effect on osteoblasts during periodontal regeneration. PMID- 11491634 TI - Comparison of the first file that fits at the apex, before and after early flaring. AB - The purpose of this study was to compare the first file that fits to the apex (FFFA) in each canal before and after early flaring to analyze if the size of file to fit to the apex would increase after flaring. One hundred mesial canals of lower first and second molars with complete apical formation and patent foramens were selected. The samples were randomly divided into two groups of 50 canals each. A file was fit to the apex in each canal and that size recorded. Radicular flaring was completed using Gates-Glidden drills in group 1 and Rapid Body Shapers in group 2. After flaring a file was again fit to the apex in the same manner as before and its size recorded. The mean diameter of FFFA before flaring (file diameters in mm x 10(-2)) was 14.46 (+/-4.12) and after 23.3 (+/ 7.2) for group 1 (p < 0.001), whereas in group 2 the mean diameter of FFFA was 17.2 (+/-4.96) before and 25.6 (+/-6.36) after (p < 0.001). A Wilcoxon t test indicates a significant difference (p < 0.001) between the diameter of FFFA before and after flaring in both groups. The increase in diameter was approximately two file sizes for both groups. From this observation it is concluded that early radicular flaring increases the size file that is snug at the apex, and awareness of that difference gives the clinician a better sense of canal size. Early flaring of the canal provides better apical size information and with this awareness, a better decision can be made concerning the appropriate final diameter needed for complete apical shaping. PMID- 11491635 TI - Effect of penicillin on postoperative endodontic pain and swelling in symptomatic necrotic teeth. AB - The purpose of this prospective, randomized, double-blind, placebo-controlled study was to determine the effect of penicillin on postoperative pain and swelling in symptomatic necrotic teeth. Forty-one emergency patients participated and each had a clinical diagnosis of a symptomatic necrotic tooth with associated periapical radiolucency. After endodontic treatment patients randomly received a 7-day oral dose (twenty-eight 500 mg capsules to be taken every 6 h) of either penicillin or a placebo control in a double-blind manner. Patients also received ibuprofen; acetaminophen with codeine (30 mg); and a 7-day diary to record pain, percussion pain, swelling, and number and type of pain medication taken. The majority of patients with symptomatic necrotic teeth had significant postoperative pain and require analgesic medication to manage this pain. The administration of penicillin postoperatively did not significantly (p > 0.05) reduce pain, percussion pain, swelling, or the number of analgesic medications taken for symptomatic necrotic teeth with periapical radiolucencies. PMID- 11491636 TI - Evaluation of success rate of endodontic treatment performed by students with stainless-steel K-files and nickel-titanium hand files. AB - A previous study by our group found nickel-titanium (NiTi) 0.02 hand files maintained the original canal shape better than similar stainless-steel K-files (SS-K). Inexperienced dental students used both file types on molar teeth. The purpose of this study was to compare the 1-yr success rate of endodontic treatment of the same teeth used in our previous study. Twelve-month follow-up radiographs were compared with the immediate follow-up radiographs. Both sets of radiographs were taken with the same customized stent. Quantification of osseous changes using digital imaging was used. Thus, a reliable numerical estimation (densitometric ratio) of disease and healing processes could be established. Sixty-seven percent of the patients returned for the 12-month radiographs (19 NiTi vs. 21 SS-K). Immediate postoperative periapical status was found to be similar (p > 0.05). Teeth instrumented with the NiTi files demonstrated a higher mean change in densitometric ratio, compared with SS-K files (p < 0.05). Further tests of success (values: > or =0) and failure (value: < or =0) with the Fisher exact test showed more success (decreasing radiographic density) with NiTi files and more failures (increasing radiographic density) with SS-K type files (p < or = 0.03). This study indicates that maintaining the original canal shape after instrumentation leads to a better prognosis of endodontic treatment. PMID- 11491637 TI - Fiberoptic endoscope usage for intracanal visualization. AB - A new method of magnified intracanal visualization using a fiberoptic endoscope has been developed for use in conventional endodontic treatment. The 0.7 mm flexible fiberoptic endoscope allows the endodontist to view canal morphology and biomechanical canal preparations, and helps to identify fractures, accessory canals, and remnant pulpal tissue. PMID- 11491638 TI - An inexpensive device for transillumination. AB - Patients with cracked-tooth syndrome often present with a chief complaint of vague pains while eating that they cannot localize, making the diagnosis difficult. Frequently practitioners are unable to discover the cause of their symptoms, so treatment is delayed, which can result in apical propagation of the crack, leading to unrestorable vertical root fracture. Transillumination is a valuable aid to diagnosis, but suitable diagnostic lights are difficult to obtain, most being cumbersome and expensive. A portable inspection/bore light that serves as an ideal device for transillumination is available for approximately $10. PMID- 11491639 TI - Clinical application of a fiber-reinforced post system. AB - The tooth structure of an endodontically treated tooth was restored with a core on a post, both prefabricated with fiber-reinforced polymer-ceramic material (Sculpture/FibreKor) and subsequently with a metal-free crown. The tooth restoration is described in detail. It includes root canal preparation, polyvinyl siloxane impression taking, laboratory fabrication of the core on the post, adhesive cementation of the post-and-core system in the root canal, fabrication and cementation of the restorations, adjustment, and finishing. The success of this metal-free system is based on its increased flexural and tensile strength. Its application is safe, effective, and reliable. The high-quality aesthetics of the restorations is due to the translucency of the post-and-core material. PMID- 11491640 TI - Knowledge and advocacy--the keys to strengthening endodontics for the future. PMID- 11491641 TI - Quantitation of catecholamines in inflamed human dental pulp by high-performance liquid chromatography. AB - Catecholamines may play an important role in the control of intrapulpal pressure as mediators of vasoconstriction. A baseline level of catecholamines (dopamine, epinephrine, and norepinephrine) in the uninflamed human dental pulp was previously reported using high-performance liquid chromatography. The purpose of this study was to compare the level of catecholamines present in the inflamed human dental pulp with the baseline level established in virgin teeth. Twelve uninflamed pulps were analyzed as a control and to validate previous findings. Pulp tissue was obtained from 10 vital and inflamed teeth requiring endodontic treatment. Selective criteria for each patient included: absence of systemic disease, medications, and allergies; a vital response to ice, heat, and electric pulp tests; and periodontal probing < or = 3 mm. A prior history of pain associated with the tooth was an additional criterion for inflamed pulps. To avoid the presence of an exogenous catecholamine, local anesthesia without epinephrine was administered. Dopamine, epinephrine, and norepinephrine were chemically extracted and analyzed by high-performance liquid chromatography with ultraviolet detection. Catecholamine levels found to be present in the pulp during inflammation were greater than the baseline level established in uninflamed pulp tissue. PMID- 11491642 TI - Viable bacteria in root dentinal tubules of teeth with apical periodontitis. AB - Two sets of teeth with apical periodontitis were collected at different geographic locations to study the identity of bacteria left in the root dentinal tubules. Root dentin of 20 of these teeth was cultured from three locations between pulp and cementum (A, B, and C). In addition dentin from eight teeth was examined histologically. Using the culturing technique bacteria were found in 77% of the dentin samples from set 1 (Amsterdam) and in 87.5% of the dentin samples from set 2 (Glasgow). At greater distance, in layer C, from the pulp bacteria were found in 62% (13 of 21) of the dentin samples. Twenty-three percent (3 of 13) of set 1 and 25% (2 of 8) of set 2 contained >50,000 colony-forming units/mg of dentin in layer C. In layers closer to the pulp higher numbers of anaerobic bacteria and gram-positive rods were found, as well as a larger number of bacterial species. Histological sections showed bacterial penetration in dentinal tubules in 5 of 8 teeth. In the other three teeth where the colony-forming units/mg recovered was <10,000, no histological signs of tubule penetration was seen. It seems clear that, in more than half of the infected roots, bacteria are present in the deep dentin close to the cementum and that anaerobic culturing of dentin is more sensitive than histology to detect these bacteria. PMID- 11491643 TI - Cytotoxicity of halothane on human gingival fibroblast cultures in vitro. AB - Recently halothane has been reported to be the most suitable alternative to chloroform in dissolving gutta-percha. Periapical tissue toxicity of halothane is not completely known. In this study gutta-percha dissolved by halothane was evaluated with the almar blue dye assay using human gingival fibroblast cultures. The cytotoxic effects of halothane on human gingival fibroblasts depended on the exposure dose, frequency, and duration. A reduced concentration and smaller amount of gutta-percha solvents may minimize the cytotoxic effects on host tissues. PMID- 11491644 TI - Adhesion of glass-ionomer cement sealers to bovine dentin conditioned with intracanal medications. AB - This in vitro study assessed the adherence of glass-ionomer cement (GIC) root canal sealers to dentin conditioned by three endodontic intracanal medications. Three GIC sealers were used: (i) Ketac-Endo; (ii) KT-308, an experimental GIC sealer; and (iii) ZUT, a combination of KT-308 and a silver-containing zeolite (0.2% by weight). Superficial dentin of 120 bovine incisor crowns was used as a substrate. The dentin was irrigated with 2.6% NaOCI for 30 s and then blotted dry. One of the following conditioning media (n = 30) was maintained in contact with the dentin for 7 days: (i) Ca(OH)2 paste; (ii) chlorhexidine gluconate (CHX) liquid 0.12%; (iii) formocresol (FML) liquid; (iv) distilled water (dH2O) used as control. The GIC sealers were applied to the conditioned dentin, bench set for 90 min, stored in 100% humidity at 37 degrees C for 48 h, then tested to failure for shear bond strength (MPa) in an Instron machine. In the ZUT specimens, the shear bond strength did not differ significantly among those conditioned with Ca(OH)2, CHX, FML, and dH2O. For KT-308, the mean scores were significantly lower (p < 0.05) after conditioning with CHX than with dH2O. For Ketac-Endo, the mean scores were significantly lower after conditioning with Ca(OH)2 and FML than with dH2O (p < 0.05). Furthermore Ketac-Endo demonstrated significantly lower (p < 0.05) shear bond strength than KT-308 or ZUT to the dentin conditioned with Ca(OH)2 or FML. The results suggest that intracanal medications differentially influence the adhesion of various GIC sealers to root canal dentin. PMID- 11491645 TI - Heat shock protein 27 expression in the epithelium of periapical lesions. AB - Heat shock protein (HSP) 27 is a member of the small HSP family that plays a part in the regulation of epithelial cell growth and differentiation, wound healing, apoptosis and cell protection against inflammatory cytotoxicity mediators. Thus the expression of HSP27 was investigated immunohistochemically in periapical granulomas with epithelial rests of Malassez and in radicular cysts. Anti-HSP27 mouse monoclonal antibody and peroxidase-labeled streptavidin-biotin standard technique were used to study the expression of HSP27. Proliferating epithelial cell rests, and islands of epithelium and epithelial lining of microcysts strongly reacted throughout all layers, whereas radicular cysts epithelial lining presented mainly a moderate suprabasal staining pattern. However both the proliferating epithelial cell rests and the radicular cysts shared an over expression of HSP27 immunostaining intensity in coincidence with the presence of local infiltration of immune cells. HSP27 may play several roles in periapical lesions that include contributing to the migration of epithelial cell rests and an increased resistance both to necrotic and apoptotic cell deaths. PMID- 11491646 TI - Comparison of fracture rate, deformation rate, and efficiency between rotary endodontic instruments driven at 150 rpm and 350 rpm. AB - Two groups of 30 mature molars each (S and F) were instrumented with Profile .04 taper Series 29 rotary instruments. Group S molars were instrumented at 150 rpm and group F at 350 rpm. The number of fractures, deformed files, and instrumentation time were recorded for each tooth. No instrument fractures occurred in either group. In group S the mean deformation rate and instrumentation time were 1.1 deformed files and 8.0 min per molar. In group F they were 0.57 deformed files and 4.6 min per molar. Both differences were significant (p < 0.05). The results indicate that Profile .04 taper Series 29 rotary instruments should be used at 350 rpm to nearly double efficiency and halve the deformation rate, compared with 150 rpm. Because no instrument fractures occurred while instrumenting 60 mature molars, both speeds should be considered safe. PMID- 11491647 TI - Dentin barrier test with transfected bovine pulp-derived cells. AB - Growth kinetics of SV40 large T-antigen-transfected bovine pulp-derived cells on dentin were investigated. These cells were used in a dentin barrier test device, and the system was evaluated by testing a set of dental filling materials. Cells (120 cells/mm2) were seeded on dentin slices and incubated for up to 21 days. Cell proliferation was recorded using MTT assay. For cytotoxicity tests 3500 cells/mm2 were seeded on dentin discs, which were then incorporated into the dentin barrier test device. After 72 h preincubation test materials were applied. After a 24 h exposure with or without perfusion of the pulpal part of the test device, cell survival was evaluated using MTT assay. The cells revealed similar growth kinetics on dentin slices and on tissue culture plates. In cytotoxicity tests the cells were more sensitive toward the test materials than previously used three-dimensional cultures of human foreskin fibroblasts and as anticipated from clinical experience. Further improvement is expected by using three dimensional cultures of pulp-derived cells. PMID- 11491648 TI - Redox-modulating gene therapies for human diseases. AB - Baseline levels of reactive oxygen species (ROS) are generated as an integral component of cellular function. Under certain conditions, e.g., the presence of an elevated concentration of transition metal (Fe/Cu) ions, drug metabolism, or ischemia-reperfusion, ROS generation is exaggerated to an extent that overwhelms cellular antioxidant defenses and results in oxidative stress. Oxidative stress has been characterized by the assessment of oxidative damage to cellular components, e.g., protein, lipid, and nucleic acid. More recent studies have determined that at a concentration much below that required for inflicting oxidative damage, ROS may serve as cellular second messengers through the regulation of numerous signal transduction pathways. For this reason, much of the current medical focus in this area has been directed toward the understanding of redox-driven physiological and pathophysiological processes in the cell. The goal of such research is to formulate effective strategies for manipulating the cellular redox environment in a manner that is beneficial for restoring normal cell functions in the setting of disease. PMID- 11491649 TI - Modulation of redox signal transduction pathways in the treatment of cancer. AB - Reactive oxygen species (ROS)-mediated damage to DNA is associated with induction of stress-activated protein kinases leading to secondary and tertiary effects on the nuclear matrix, cytoplasmic transport mechanisms, and altered mitochondrial and cell membranes. The cellular defenses against ROS damage are associated with up-regulation of gene products that can significantly alter cell biology, including antiapoptotic Bax family proteins and inflammatory proteins. Altered cell integrity can occur either directly or by indirect paracrine and juxtacrine interactions within tissues. Previous approaches toward therapeutic intervention against ROS damage have included administration of radical scavenger compounds, use of novel drugs that increase cellular production of constitutive antioxidants, or pharmacologic agents that modify the intracellular transport of antioxidants. Strategies to modify the cellular effects of ROS in hyperbaric oxygen injury to the lung, reperfusion injury to transplanted organs, and cancer have led to novel approaches of gene therapy in which the transgenes for antioxidant proteins can be expressed in specific tissues. Reducing tissue damaging effects of ROS may have relevance to cancer patients by ameliorating normal tissue damage from ionizing irradiation therapy, photodynamic therapy, and cancer chemotherapy. PMID- 11491650 TI - Potential mechanisms for the inhibition of tumor cell growth by manganese superoxide dismutase. AB - Studies from many laboratories have shown that overexpression of manganese superoxide dismutase (MnSOD) inhibits the growth of numerous tumor cell types. The inhibition of tumor cell growth can be attributed to the increase in the steady-state levels of H2O2 as a result of the increased dismuting activity of MnSOD. Here we demonstrate that overexpression of MnSOD enhances the activity of the superoxide (O2*-)-sensitive enzyme aconitase, decreases the intracellular GSH/GSSG ratio, and dose-dependently inhibits pyruvate carboxylase activity. Thus, alterations in the steady-state concentrations of mitochondrial O2*- and H2O2 as a result of MnSOD overexpression can alter the metabolic capacity of the cell leading to inhibition of cell growth. Furthermore, we propose that MnSOD overexpression can modulate the activity of nitric oxide (*NO) by preventing its reaction with O2*-. This hypothesis suggests that the redox environment of the mitochondria can be altered to favor the activity of *NO rather than peroxynitrite (ONOO-) and may explain the enhanced toxicity of *NO-generating compounds toward MnSOD-overexpressing cell lines. These findings indicate that therapeutic strategies targeted at overexpressing MnSOD in tumor tissue may be more effective when used in combination with agents that deplete the oxidant buffering and enhance the *NO-generating capacity of the tumor and host, respectively. PMID- 11491651 TI - Constitutive activation of transcription factor AP-2 is associated with decreased MnSOD expression in transformed human lung fibroblasts. AB - Activator protein-2 (AP-2) is a transcription factor with transactivating and transrepressing potential in different promoter contexts. AP-2 contains seven cysteines, and its in vitro DNA binding activity is redox-sensitive. Superoxide dismutase-2 (SOD2), which encodes the antioxidant enzyme manganese superoxide dismutase (MnSOD), is a putative tumor suppressor gene whose loss of expression is associated with the malignant phenotype. SOD2 promoter mutations that generate new AP-2 sites are associated with loss of MnSOD expression in cancer cells. In the current study, we have identified an inverse expression pattern between AP-2 and MnSOD in normal versus transformed human cells. MRC5 cells are a normal human lung fibroblast cell strain that is mortal and senesces after a certain number of passages in vitro. MRC5-VA is a simian virus transformed variant of MRC5. We determined the levels of expression of MnSOD and AP-2 in these two cell types at the levels of mRNA, protein, and activity. Our results indicated that MnSOD expression was significantly decreased in MRC5-VA cells compared with MRC5 cells at each level of investigation, whereas AP-2 showed an opposing pattern of expression and DNA binding activity. These results suggest that AP-2 may participate in the mechanism(s) underlying decreased expression of SOD2 in transformed cells. PMID- 11491652 TI - Gene transfer of extracellular superoxide dismutase to atherosclerotic mice. AB - Clinical and epidemiological studies have provided circumstantial evidence that oxidized low-density lipoprotein (LDL) and antioxidants are involved in the pathogenesis of atherosclerosis. Superoxide dismutases (SODs) have been shown in vitro to protect LDL from deleterious effects of superoxide anions. In the present study, we have used adenoviral gene transfer to determine effect of extracellular SOD (EC-SOD) on atherogenesis in LDL receptor -/- mice. Intravenous administration of EC-SOD adenovirus (2 x 10(9) plaque forming units) into tail vein targeted transgene mainly to liver and induced a 3.5- to sevenfold increase in plasma total SOD activity. EC-SOD was secreted into circulation for 2-3 weeks mostly in a truncated B-form, suggesting that endogenous proteolytic mechanisms control the level and distribution of the enzyme. Therapeutic potential was determined by measuring plasma resistance against copper oxidation and analyzing atherosclerotic lesion areas in aortas of LDL receptor -/- mice. Mice were kept on a cholesterol diet for 10 weeks before gene transfer and 3 or 6 weeks after the gene transfer. Results showed a tendency for a reduction in the overall lesion area after EC-SOD gene transfer as compared with LacZ transduced control mice, but the difference did not reach statistical significance. It is concluded that short-term overexpression of EC-SOD in vivo does not affect atherogenesis in LDL receptor -/- mice. PMID- 11491653 TI - An essential role of the antioxidant gene Bcl-2 in myocardial adaptation to ischemia: an insight with antisense Bcl-2 therapy. AB - Reperfusion of ischemic myocardium results in apoptotic cell death, which can be blocked by adapting the heart to ischemic stress induced by cyclic episodes of brief periods of ischemia and reperfusion. In concert, the antiapoptotic gene bcl 2 is decreased by ischemia/reperfusion, but increased in the ischemically adapted myocardium. To examine if bcl-2 plays a crucial role in cardioprotection, adaptive cardioprotection was further examined in the hearts treated with antisense bcl-2 oligodeoxynucleotides (ODN). Isolated Langendorff-perfused rat hearts were divided into three groups: control (perfused with Krebs-Henseleit bicarbonate buffer for 210 min); 30-min ischemia followed by 2-h reperfusion; ischemic adaptation followed by 30-min ischemia and 2-h reperfusion. The last (adapted heart) group was subdivided into another two groups: one was transfected 48 h earlier with antisense bcl-2 ODN, whereas the other group was transfected with sense bcl-2 ODN. Cardioprotection was examined by determining cardiomyocyte death due to necrosis and apoptosis. Antisense gene therapy almost completely abolished bcl-2 protein expression in the hearts. Bcl-2 mRNA was down-regulated in the ischemic/reperfused heart, but up-regulated in the adapted myocardium. Adapted myocardium showed decreased infarct size and reduced number of apoptotic cardiomyocytes. Ischemia/reperfusion resulted in increased oxidative stress as evidenced by increased malonaldehyde formation. Adapted myocardium had a reduced amount of malonaldehyde. Antisense bcl-2 ODN completely abolished the cardioprotective effects of adaptation by eliminating the antideath signal of bcl 2. In concert, reduced oxidative stress in the adapted myocardium no longer persisted. The results suggest an antioxidant role of bcl-2 that appeared to be essential for the cardioprotection achieved by ischemic adaptation. PMID- 11491654 TI - GPx-1 gene delivery modulates NFkappaB activation following diverse environmental injuries through a specific subunit of the IKK complex. AB - Numerous environmental stimuli alter cell functions by the induction of intracellular reactive oxygen species, such as superoxide and hydrogen peroxide (H2O2). These redox alterations can change the activity of kinases and phosphatases responsible for controlling intracellular signal transduction cascades important in determining how cells react to their environment. One such well known pathway includes nuclear factor-kappaB (NFkappaB); however, the exact redox-sensitive factors important in controlling H2O2-mediated activation of NFkappaB remain unclear. In the present study, we have investigated how intracellular clearance of H2O2, using a recombinant adenovirus expressing glutathione peroxidase-1 (GPx-1), modulates NFkappaB activation following UV irradiation, tumor necrosis factor-alpha, or H2O2 treatment of MCF-7 cells. Findings from these studies demonstrate that GPx-1 overexpression can down regulate NFkappaB DNA binding, and transcriptional activation of an NFkappaB dependent luciferase reporter, to varying extents following these environmental stimuli. Studies using dominant negative adenoviral vectors expressing IKKalpha(KM) and IKKbeta(KA) suggest that GPx-1-mediated H2O2 clearance appears to preferentially inhibit the activity of IKKalpha, but not IKKbeta. These studies demonstrate for the first time that redox regulation of NFkappaB activation by intracellular H2O2 may be specific for a unique subunit in the IKK complex. Such findings suggest that IKK kinases or IKK phosphatases may have unique redox-regulated components. These studies have shed mechanistic insight into the potential application of redox-modulating gene therapies aimed at altering NFkappaB activation following environmental injury. PMID- 11491655 TI - Gene therapeutic approaches to oxidative stress-induced cardiac disease: principles, progress, and prospects. AB - Heart and vascular diseases continue to rank among the most frequent and devastating disorders to affect adults in many parts of the world. Increasing evidence from a variety of experimental models indicates that reactive oxygen species can play a key role in the development of myocardial damage from ischemia/reperfusion, the development of cardiac hypertrophy, and the transition of hypertrophy to cardiac failure. The recent dramatic increase in availability of genomic data has included information on the genetic modulation of reactive oxygen species and the antioxidant systems that normally prevent damage from these radicals. Nearly simultaneously, progressively more sophisticated and powerful methods for altering the genetic complement of selected tissues and cells have permitted application of gene therapeutic methods to understand better the pathophysiology of reactive oxygen species-mediated myocardial damage and to attenuate or treat that damage. Although exciting and promising, gene therapy approaches to these common disorders are still in the experimental and developmental stages. Improved understanding of pathophysiology, better gene delivery systems, and specific gene therapeutic strategies will be needed before gene therapy of oxyradical-mediated myocardial damage becomes a clinical reality. PMID- 11491656 TI - Approaches and limitations to gene therapy for mitochondrial diseases. AB - Mitochondrial dysfunction may be caused by mutations in either the nuclear and/or the mitochondrial genome. Since 1988, mitochondrial DNA mutations have been linked to retinopathies, myopathies, neurodegenerative diseases, and possibly normal aging. Adequate drug therapies for these disorders have yet to be discovered. Therefore, gene therapy must be considered as a possible alternative. In this review, we will discuss the possibilities and the problems associated with gene therapy for mitochondrial disorders. PMID- 11491657 TI - Anticancer therapy by overexpression of superoxide dismutase. AB - Cancer cells are in general low in the enzymatic activities of both manganese containing (MnSOD) and copper- and zinc-containing superoxide dismutase. We have hypothesized that part of the tumor cell phenotype is due to this loss of enzymatic activity. To test this hypothesis, we have overexpressed MnSOD via plasmid and adenovirus transfection in various cancer cell types and have shown tumor suppression. This tumor suppression is via a noncytotoxic mechanism and probably occurs due to cell-cycle perturbations. We have also shown that MnSOD overexpression causes the anticancer drug 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) to have increased cytotoxicity. Our hypothesis for the mechanism of action of this combination is that overexpression of MnSOD leads to increased peroxide levels and that BCNU inhibits peroxide removal. We currently are investigating the use of adenovirus MnSOD plus BCNU in the treatment of cancer. Results thus far are consistent with the idea that we can use the alterations in antioxidant enzymes observed in cancer cells to therapeutic advantage. PMID- 11491658 TI - Osmotic stress-mediated activation of RET kinases involves intracellular disulfide-bonded dimer formation. AB - We showed that osmotic stress induces activation of c-RET and second-set activation of constitutively activated RET-MEN2B. A few percentage of RET proteins normally formed disulfide-bonded dimers in the cell, and osmotic stress promoted formation of these dimers. The disulfide-bonded dimers displayed higher levels of autophosphorylation and catalytic activity per molecule than did monomers. Osmotic stress also promoted activation and disulfide-bonded dimerization of the extracellular domain-depleted mutant RET (RET-PTC-1), suggesting that the target amino acid(s) for dimerization is located intracellularly rather than in the cysteine-rich region of the extracellular domain. In the mutant c-RET and RET-PTC-1 in which Cys987 of c-RET or Cys376 of RET-PTC-1 was replaced with Ala, the levels of intrinsic kinase activity were greatly reduced and barely increased in response to osmotic stress. Correspondingly, the Cys376-defective RET-PTC-1 did not form any demonstrable levels of dimers even after exposure to osmotic stress. In contrast, another RET PTC-1 mutant that had a replacement of Cys365 with Ala mostly behaved like parental RET-PTC-1. These results suggest that Cys987 of c-RET or Cys376 of RET PTC-1 plays a crucial role in maintenance and promotion of dimerization and activation of the RET kinases. PMID- 11491659 TI - Alterations in hepatic kinase activity following whole body gamma-irradiation of mice. AB - The chronological activation of the signaling molecules following whole body gamma-irradiation was investigated in mouse liver. The activity of two kinases, tyrosine kinase and protein kinase C (PKC), was found to respond differently to gamma-irradiation. Tyrosine kinase was found to respond to much lower doses of irradiation (10 cGy), whereas PKC was found to be activated at comparatively higher doses (3 Gy). Tyrosine kinase showed a sharp activation at 30 min and then a decline to normal values at 1 h. Activation of PKC was apparent at as early as 15 min of irradiation and showed a maximal increase at 30 min. This was followed by a decline to normal values at 1 h. The response of the whole organ was found to be different from that of reported effects on a single cell. These results suggest that the data obtained from the single-cell studies would have limited application in the experiments involving the whole animal. Interruption of these signals at various steps is currently being used to manipulate tumor response to radiotherapy. In such cases, the difference in response of a single cell and a whole animal must be considered. PMID- 11491661 TI - Nitric oxide and superoxide radical production by human mononuclear leukocytes. AB - Human mononuclear cells (90% lymphocytes, 9% monocytes, and 1% polymorphonuclear leukocytes) produced spontaneously in resting state 0.11+/-0.01 nmol of nitric oxide (NO)/min/10(6) cells and 0.25+/-0.02 nmol of superoxide anion (O2 )/min/10(6) cells, as primary products. When these cells were stimulated with phorbol 12-myristate 13-acetate (PMA), the NO and O2- production increased by 82% and 204% to 0.25+/-0.02 nmol of NO/min/10(6) cells and 0.76+/-0.12 nmol of O2 /min/10(6) cells, respectively. Oxygen uptake reasonably accounted for the sum of the rates of NO and hydrogen peroxide (H2O2), the latter calculated as 0.5 O2- production, in nonstimulated and in PMA-stimulated cells. H2O2 and peroxynitrite formation were detected and measured as secondary products of the primary products O2- and NO. An original assay to determine H2O2 steady-state concentration and production rates is described. The determined production rates of the involved reactive species are in good agreement with known chemical equations. It is apparent that NO and O2- production by human mononuclear cells may constitute the basis of intercellular signaling and cell toxicity. PMID- 11491660 TI - Hydrogen peroxide activates NFkappaB and the interleukin-6 promoter through NFkappaB-inducing kinase. AB - Aging is associated not only with oxidant stress, but also with increased interleukin-6 (IL-6) levels. To determine if oxidative stress could contribute to the age-associated increase IL-6 expression, we exposed LNCaP prostate carcinoma cells and HeLa cervical carcinoma cells to H2O2 as an oxidant challenge. We found that H2O2 induced IL-6 expression through activation of the IL-6 promoter. Furthermore, H2O2-induced activation of the promoter was mediated through nuclear factor-kappaB (NFkappaB) secondary to H2O2-induced phosphorylation and degradation of IkappaBalpha. NFkappaB-inducing kinase (NIK) is upstream of the IkappaB kinase complex that induces IkappaBalpha degradation. Accordingly, we explored if H2O2 induces IL-6 expression through NIK. In addition to H2O2 inducing NIK autophosphorylation, transfection of LNCaP cells with a dominant negative NIK diminished H2O2-mediated NFkappaB and IL-6 promoter activity. Taken together, these results demonstrate that H2O2 induces the IL-6 promoter by activating NFkappaB through NIK. These data provide a candidate mechanism through which oxidant challenge induces IL-6 gene expression with age. PMID- 11491662 TI - Redox control of mitochondrial functions. AB - Redox reactions and electron flow through the respiratory chain are the hallmarks of mitochondria. By supporting oxidative phosphorylation and metabolite transport, mitochondrial redox reactions are of central importance for cellular energy conversion. In the present review, we will discuss two other aspects of the mitochondrial redox state: (i) its control of mitochondrial Ca2+ homeostasis, and (ii) the intramitochondrial formation of reactive oxygen or nitrogen species that strongly influence electron flow of the respiratory chain. PMID- 11491663 TI - Free radical-dependent Ca2+ signaling: role of Ca2+-induced Ca2+ release. AB - Previously we have shown that Fe3+/ascorbate-induced Ca2+ release from scallop sarcoplasmic reticulum (SR) is due to Ca2+-channel gating by free radicals. This study is aimed at demonstrating that Ca2+-induced Ca2+ release (CICR) plays a role in this kind of Ca2+ release. Scallop SR vesicles were incubated with fluo-3 and exposed to Fe3+/ascorbate. Fluorimetric recordings showed massive Ca2+ release, with maximum rate and 50% release occurring at 30 min after exposure. Conversely, the use of the probe for reactive oxygen species dihydrorhodamine or the assay of malondialdehyde allowed oxyradical production to be traced for approximately 5 min only. Hence, although Ca2+ release started just after exposure to Fe3+/ascorbate, most release occurred after free radical exhaustion. Ruthenium red addition after Fe3+/ascorbate slowed down the Ca2+ release, whereas cyclic adenosine 5'-diphosphoribose addition accelerated it, indicating that the free radical-induced Ca2+ release from SR vesicles triggers a mechanism of CICR that dramatically increases the initial effect. PMID- 11492952 TI - Brain angiotensin and body fluid homeostasis. AB - Angiotensinogen, the precursor molecule of the peptides angiotensin I, II, and III, is synthesized in the brain and the liver. Evidence is reviewed that angiotensin II, and possibly angiotensin III, that are generated within the brain act within neural circuits of the central nervous system to regulate body fluid balance. Immunohistochemical studies in the rat brain have provided evidence of angiotensin-containing neurons, especially in the hypothalamic paraventricular nucleus, subfornical organ, periventricular region, and nucleus of the solitary tract, as well as in extensive angiotensin-containing fiber pathways. Angiotensin immunoreactivity is observed by electron microscope in synaptic vesicles in several brain regions, the most prominent of these being the central nucleus of the amygdala. Neurons in many parts of the brain (lamina terminalis, paraventricular and parabrachial nuclei, ventrolateral medulla, and nucleus of the solitary tract) known to be involved in the regulation of body fluid homeostasis exhibit angiotensin receptors of the AT(1) subtype. Pharmacological studies in several species show that intracerebroventricular administration of AT(1) receptor antagonist drugs inhibit homeostatic responses to the central administration of hypertonic saline, intravenous infusion of the hormone relaxin, or thermal dehydration. Responses affected by centrally administered AT(1) antagonists are water drinking, vasopressin secretion, natriuresis, increased arterial pressure, reduced renal renin release, salt hunger, and thermoregulatory adjustments. We conclude that angiotensinergic neural pathways in the brain probably have an important homeostatic function, especially in regard to osmoregulation and thermoregulation, and the maintenance of arterial pressure. PMID- 11492953 TI - Selective brain cooling in mammals and birds. AB - Artiodactyls and felids have a carotid rete that can cool the blood destined for the brain and consequently the brain itself if the cavernous sinus receives cool blood returning from the nose. This condition is usually fulfilled in resting and moderately hyperthermic animals. During severe exercise hyperthermia, however, the venous return from the nose bypasses the cavernous sinus so that brain cooling is suppressed. This is irreconcilable with the assumption that the purpose of selective brain cooling (SBC) is to protect the brain from thermal damage. Alternatively, SBC is seen as a mechanism engaging the thermoregulatory system in a water-saving economy mode in which evaporative heat loss is inhibited by the effects of SBC on brain temperature sensors. In nonhuman mammals that do not have a carotid rete, no evidence exists of whole-brain cooling. However, the surface of the cavernous sinus is in close contact with the base of the brain and is the likely source of unregulated regional cooling of the rostral brain stem in some species. In humans, the cortical regions next to the inner surface of the cranium are very likely to receive some regional cooling via the scalp-sinus pathway, and the rostral base of the brain can be cooled by conduction to the nearby respiratory tract; mechanisms capable of cooling the brain as a whole have not been found. Studies using conventional laboratory techniques suggest that SBC exists in birds and is determined by the physical conditions of heat transfer from the head to the environment instead of physiological control mechanisms. Thus except for species possessing a carotid rete, neither a coherent pattern of SBC nor a unifying concept of its biological significance in mammals and birds has evolved. PMID- 11492954 TI - Lactate release, concentration in blood, and apparent distribution volume after intense bicycling. AB - To study the release of lactate from muscle and its relationship to the blood lactate concentration during and after intense bicycling, young men cycled at 5.5 W kg(-1) body mass for 2 min to exhaustion or stopped after 1 min (nonexhaustive ride). The leg's release of lactate during and after each ride was taken from the measured blood flow and lactate concentrations in arterial and femoral-venous blood. Muscle biopsies were taken in separate experiments and analyzed for lactate. During the bicycling, 6 to 10% of the lactate produced was released to the blood. During exercise and for the first few minutes after, the rate of lactate release did not differ between 2 min exhaustive and 1 min nonexhaustive bicycling. The integrated release (exercise plus recovery) for the 1 min bicycling was 60 to 80% of the corresponding value of the 2 min exhaustive bicycling. In the late recovery, the blood lactate concentration was 3 to 5 times higher after 2 min exhaustive bicycling than after the 1 min nonexhaustive bicycling. There was thus a mismatch between the amount of lactate released and measured concentration in blood, reflecting a smaller distribution volume after the exhaustive bicycling. The blood lactate concentration may therefore not be a good measure of the lactate production and anaerobic energy release during bicycling. PMID- 11492955 TI - Expression of vascular endothelial growth factor and its receptors in the developing rat lung. AB - We examined the regulation of the expression of vascular endothelial growth factor (VEGF) and its specific receptors, fetal liver kinase receptor (Flk-1), and fms-like tyrosine kinase receptor (Flt-1) during formation of the capillary network in the developing rat lung. An immunohistochemical study of lung tissue from 19- and 21-d-old fetuses and 1-, 3-, 5-, 7-, and 14-d-old animals revealed that the level of expression of both VEGF and Flk-1 is significantly higher before birth (p < 0.0001) than after. Increased expression of Flt-1 on the first day after birth (p < 0.0001) suggests that this receptor might play an important role in capillary growth in the perinatal period. Immunostaining also revealed the colocalization of VEGF, Flt-1, and Flk-1 in endothelial cells of the lung capillaries at the ultrastructural level. The present studies revealed that VEGF and its two receptors are upregulated during the development of capillaries in the fetal and newborn rat lung. PMID- 11492956 TI - Cross-bridge and calcium behavior in ferret papillary muscle in different thyroid states. AB - X-ray diffraction studies were made using synchrotron radiation on ferret right ventricular papillary muscle under three different thyroid states: euthyroidism, hyperthyroidism, and hypothyroidism. The latter two states were induced by treatment with L-thyroxine and methimazole, respectively. The X-ray equatorial reflections were recorded at a time resolution of 10 ms to study the mass movement of myosin cross-bridges from thick to thin filaments. The myosin isomer content was measured by gel electrophoresis which showed that V3 isomer was predominant in euthyroid muscle and 27% of myosin was V1 isomer in hyperthyroid muscle. The intracellular free Ca concentration was measured by using the aequorin method. The peak Ca concentration was similar in all three states, but in the hypothyroid state the time-to-peak was longer and the decay was slower. The time-to-peak of twitch tension was shorter in hyperthyroidism and longer in hypothyroidism than in euthyroidism. The different time courses of twitch tension in different thyroid states accompanied a cross-bridge movement which closely followed the tension development. In hyperthyroidism, the cross-bridge movement significantly preceded tension development, suggesting that hyperthyroid myosin (V1) has a longer latency period between the shift to the vicinity of the thin filament and force development. PMID- 11492957 TI - Involvement of different activator Ca(2+) in the rate-dependent stretch-induced contractions of canine basilar artery. AB - Stretch evoked a contraction in a rate-dependent manner in canine basilar artery; slow stretch at rates less than 3 mm/s produced no active tension, whereas quick stretch at rates over 5 mm/s did. Large conductance Ca(2+)-activated K(+) channel blockers, including charybdotoxin, iberiotoxin, and tetraethylammonium (TEA) sensitized the basilar artery to mechanical stimulation. TEA shifted the stretch rate-tension relationship toward the left. Thus, in the presence of TEA, the slow stretch (0.1-3 mm/s) could increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) and active tension. The contraction in response to slow stretch (1 mm/s) was abolished by nicardipine and Gd(3+). Quick stretch (100 mm/s) increased [Ca(2+)](i) and active tension, both of which were partially inhibited by nicardipine or Gd(3+). The Gd(3+)-insensitive component of quick stretch-induced contraction was eliminated by thapsigargin, but not by nicardipine. Ryanodine, cyclopiazonic acid, thapsigargin, U-73122, and calphostin C also abolished the nicardipine-insensitive component of quick stretch-induced contraction. These results suggest that the slow stretch-induced contraction was exclusively dependent on the Ca(2+) influx through L-type voltage-dependent Ca(2+) channels (VDCs), whereas the quick stretch-induced contraction was dependent on Ca(2+) release from sarcoplasmic reticulum (SR) and Ca(2+) influx through L-type VDCs. PMID- 11492958 TI - Roles of alpha(1) and alpha(1)/beta subunits derived from cardiac L-type Ca(2+) channels on voltage-dependent facilitation mechanisms. AB - Strong depolarization pulses facilitate L-type Ca(2+) channels in various cell types including cardiac myocytes. The mechanisms underlying prepulse facilitation are controversial with respect to the requirements for channel subunits, cAMP dependent protein kinase, and additional anchor proteins. The properties of voltage-dependent facilitation of the L-type Ca(2+) channel was studied in recombinant cardiac alpha(1) subunits with or without cardiac beta subunit, expressed in Chinese hamster fibroblast cells. The magnitude of voltage-dependent I(Ba) facilitation in the alpha(1) subunit channel is dependent on the duration of the prepulse as well as on the interval duration between prepulse and test pulse. The characteristics of this facilitation were not affected by coexpression of the beta subunit. These results indicate that cardiac alpha(1) subunits exhibit voltage-dependent facilitation because of their own intrinsic structure, independent of any other accessory subunit or additional regulatory proteins, and that cardiac beta subunits have no essential regulatory role at the onset or continuance of the voltage-dependent facilitation. PMID- 11492959 TI - A voltage-dependent transient K(+) current in rat dental pulp cells. AB - We characterized a voltage-dependent transient K(+) current in dental pulp fibroblasts on dental pulp slice preparations by using a nystatin perforated patch recording configuration. The mean resting membrane potential of dental pulp fibroblasts was -53 mV. Depolarizing voltage steps to +60 mV from a holding potential of -80 mV evoked transient outward currents that are activated rapidly and subsequently inactivated during pulses. The activation threshold of the transient outward current was -40 mV. The reversal potential of the current closely followed the K(+) equilibrium potential, indicating that the current was selective for K(+). The steady-state inactivation of the peak outward K(+) currents described by a Boltzmann function with half-inactivation occurred at -47 mV. The K(+) current exhibited rapid activation, and the time to peak amplitude of the current was dependent on the membrane potentials. The inactivation process of the current was well fitted with a single exponential function, and the current exhibited slow inactivating kinetics (the time constants of decay ranged from 353 ms at -20 mV to 217 ms at +60 mV). The K(+) current was sensitive to intracellular Cs(+) and to extracellular 4-aminopyridine in a concentration dependent manner, but it was not sensitive to tetraethylammonium, mast cell degranulating peptide, and dendrotoxin-I. The blood depressing substance-I failed to block the K(+) current. These results indicated that dental pulp fibroblasts expressed a slow-inactivating transient K(+) current. PMID- 11492960 TI - A reciprocal regulation of Ca(2+)-activated K(+) channel by insulin and somatostatin in guinea-pig pancreatic acinar cells. AB - The effects of islet hormones, insulin and somatostatin, on the regulation of the opening of Ca(2+)-activated K(+) channels in the basolateral plasma membrane of primary cultured pancreatic acinar cells of guinea-pig were studied by cell attached single-channel recording technique. A single application of insulin or somatostatin did not influence the opening of K(+) channel. The open-state probability (p) of K(+) channel induced by the application of acetylcholine (ACh) to the bath solution was increased by insulin in the presence of ACh. The enhancement effect of insulin on the increased frequency of the channel opening was not seen when concomitantly applied with protein kinase A inhibitor, Rp-cAMPS triethylamine. Insulin increased the p value of K(+) channel, which was reversed by an application of somatostatin (Tyr-somatostatin 28). The addition of ACh followed by forskolin or dibutyryl adenosine 3',5'-cyclic monophosphate (dbcAMP) to the bath solution evoked an increase in the p value of K(+) channel. After increasing the channel opening, the addition of somatostatin reduced the p value, but not with dbcAMP. Taken together, the results suggest that insulin and somatostatin reciprocally modify the ACh-induced opening of the Ca(2+)-activated K(+) channel through a cyclic AMP-dependent signaling pathway. PMID- 11492961 TI - Laterality in direct and indirect inotropic effects of sympathetic stimulation in isolated canine heart. AB - Although sympathetic nerve stimulation is known to increase ventricular contractility, concomitant increases in heart rate (HR) make it difficult to separate its direct inotropic effect from indirect inotropic effect through a force-frequency mechanism. We stimulated the stellate ganglia in 8 isolated canine hearts with functional sympathetic nerves. Right sympathetic stimulation at 10 Hz increased ventricular end-systolic elastance (E(es)) by 95.7 +/- 7.5% (p < 0.001) and HR by 32.5 +/- 4.2% (p < 0.05). In contrast, left sympathetic stimulation at 10 Hz increased E(es) by 70.7 +/- 6.5% (p < 0.001) without significant changes in HR. Preventing the chronotropic response by fixed-rate pacing attenuated the E(es) response to right sympathetic stimulation at 5 Hz (52.0 +/- 5.1 vs. 22.8 +/- 2.8%, p < 0.001), but not to left sympathetic stimulation at 5 Hz (54.5 +/- 3.4 vs. 53.3 +/- 2.2%, NS). In the isolated canine heart, the right sympathetic nerve affected E(es) by both the direct inotropic effect and the indirect HR-dependent inotropic effect. In contrast, the left sympathetic nerve regulated E(es) primarily by its direct inotropic effect. PMID- 11492962 TI - Ca(2+)-dependent inhibition of inwardly rectifying K(+) channel in opossum kidney cells. AB - The effect of intracellular Ca(2+) on the activity of the inwardly rectifying ATP regulated K(+) channel with an inward conductance of about 90 pS was examined by using the patch-clamp technique in opossum kidney proximal tubule (OKP) cells. The activity of the inwardly rectifying K(+) channel rapidly declined with an application of ionomycin (1 microM) in the presence of 10(-6) M Ca(2+) in cell attached patches. The application of 10 microM phorbor-12-myristate-acetate (PMA) with 10(-6) M Ca(2+) reduced the K(+) channel activity. Although the channel activity was not influenced by an increase of bath Ca(2+) from 10(-7.5) to 10(-6) M, the activity was inhibited by protein kinase C (PKC, 1 U/ml) with 10(-6) M Ca(2+) in inside-out patches. The inhibitory effect of Ca(2+) with ionomycin on the channel activity was diminished by the pretreatment with a specific PKC inhibitor, GF 109203X (5 microM), in cell-attached patches. By contrast, the application of Ca(2+)/calmodulin kinase II (CaMK II, 300 pM) dramatically increased this channel activity in inside-out patches. In cell-attached patches, the addition of both GF 109203X and cyclospolin A (5 microM), a potent inhibitor of protein phosphatase 2B (calcineurin), instead stimulated the K(+) channel activity with ionomycin and 10(-6) M Ca(2+). The addition of protein phosphatase 2B (calcineurin) (2 U/ml) to the bath with calmodulin (1 microM) and Ni(2+) (10 microM) to stimulate calcineurin inhibited the channel activity in inside-out patches. Furthermore, the inhibitory effect of PKC or calcineurin on this channel activity was abolished by a removal of Ca(2+) from bath solution. These results suggest that Ca(2+)-dependent inhibitory effect on the inwardly rectifying K(+) channel in OKP cells was mainly mediated by Ca(2+)-PKC-mediated phosphorylation, and that the Ca(2+)-calmodulin-dependent phosphorylation process may be counterbalanced by the Ca(2+)-calmodulin-dependent dephosphorylation process. PMID- 11492963 TI - Anticipatory fall in core temperature in rats acclimated to heat given for various hours at a fixed daily time. AB - Rats were subjected to daily heat exposure limited to a fixed time for 10 consecutive days. An ambient temperature (T(a)) inside an animal room initially set at 24.0 degrees C was raised to 32.0 degrees C, starting at the middle of the dark phase. The high T(a) was maintained for a subsequent 1.5, 3, or 5 h and was then returned to 24.0 degrees C. After the heat exposure schedule, their core temperature significantly fell for about 30 min before and during the period when they had previously been exposed to heat without thermal stimuli. In rats, a time memory for heat exposure could be formed even when the duration of daily heat exposure was short. PMID- 11492964 TI - Effects of sarcomere length and Ca(2+) binding on h reactivity of myofilament bound troponin C in porcine skinned cardiac muscle fibers. AB - Length dependence of cardiac Ca(2+) activation is an essential component of the Frank-Starling relation. The aim of this study is to examine the length effects on the Ca(2+)-induced conformational changes of filament-bound cTnC in skinned cardiac muscle fibers. The two cysteine residues (Cys-35 and Cys-84) in the regulatory domain of cTnC allow for the attachment of conformational probes to this region. Their incorporation with the fluorescent probe, 7-diethylamino-3-[4' maleimidylphenyl]-4-methylcoumarin (CPM), was used to determine the varying cTnC conformations in cardiac fibers. The data obtained show that the length-dependent Ca(2+)-mediated conformational changes require strong-binding cross-bridges for cardiac activation. PMID- 11492965 TI - Reactive blue 2 induces calcium oscillations in HeLa cells. AB - Reactive Blue 2 (RB), which is used as an ATP receptor antagonist, induced Ca(2+) oscillations in HeLa cells. RB-induced Ca(2+) oscillations were abolished in Ca(2+)-free solution. RB, however, did not affect Ca(2+) influx measured by Mn(2+) quenching. The PLC cascade and intracellular Ca(2+) release were involved as U73122 and thapsigargin inhibited RB-induced Ca(2+) oscillations. RB enhanced a Ca(2+) response to histamine that is linked to the PLC cascade. RB may activate the PLC cascade in an extracellular Ca(2+)-dependent manner and induce Ca(2+) oscillations. PMID- 11492966 TI - Two-week, but not 1-week, hypoxic exposure enhances nitric oxide-mediated basal tone regulation in rat resistance pulmonary arteries. AB - We measured internal diameter (ID) changes in resistance and conduit pulmonary arteries of 1- and 2-week hypoxic rats and normoxic control rats in response to nitric oxide synthase (NOS) inhibitors in vivo. At 2 weeks of hypoxic exposure, the ID reduction as a result of NOS inhibition was enhanced within the resistance arteries, but not at 1 week of hypoxia. PMID- 11492967 TI - Epidemiology of primary sclerosing cholangitis. AB - Many patients suffering from primary sclerosing cholangitis (PSC) have no symptoms--or rather unspecific symptoms. Most patients have a cholestatic biochemical profile, but a specific blood test for the diagnosis of PSC is lacking. The diagnostic test (endoscopic retrograde cholangiography (ERC)) is an invasive procedure with potential complications. Also, in some patients the diagnosis of PSC is not easy, even when ERC has been performed. Therefore true incidence and prevalence data on PSC are extremely few. Nevertheless, it seems well established that the epidemiology of PSC is not the same all over the world. PSC is most often seen in Northern Europe. In this part of the world PSC is also associated with inflammatory bowel disease in most cases--and in the Nordic Countries PSC has become the primary indication for hepatic transplantation. PMID- 11492968 TI - Natural history and prognostic models in primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by inflammation and fibrosis of the intra- and extra-hepatic bile ducts. Despite the recognition of immunological and genetic alterations cited as factors in its pathogenesis, the exact cause for PSC remains unknown. Observational cohort studies, however, have demonstrated that PSC is a progressive disease culminating in liver failure or death. Natural history assessment in PSC, however, has been complicated by variable rates of disease progression and the impact of clinical symptoms upon initial presentation. The development of mathematical models by multivariable regression techniques (most notably Cox proportional hazards regression) has allowed for an improved description of overall survival on an individual basis among patients with PSC. Additionally, these models have also been employed for determining the optimal selection and timing for liver transplantation when advanced disease is imminent. PMID- 11492969 TI - Aetiopathogenesis of primary sclerosing cholangitis. AB - The aetiology and pathogenesis of PSC have not yet been clearly defined. The hypothesis that PSC is an immune mediated disease is supported by associations with HLA haplotypes, the presence of autoantibodies, increased levels of total serum immunoglobulins and the association with other autoimmune diseases. PSC does not, however, have many of the characteristics of classical autoimmunity, particularly the usual female preponderance of disease and lack of a good response to immunosuppression. Non-immune mechanisms such as bacterial infection, ischaemia and toxicity are also clearly important in the development of the disease and these factors may trigger peribiliary inflammation and cytokine induced hepatic fibrosis. PSC may be triggered in genetically susceptible individuals by toxic or infectious agents gaining access to the liver via a diseased and permeable colon. PMID- 11492970 TI - Animal models for primary sclerosing cholangitis. AB - Since the aetiopathogenesis of primary sclerosing cholangitis (PSC) in humans remains undefined, investigators have studied a variety of animal models to gain insights into immunopathogenetic mechanisms associated with obliterative fibrous cholangitis of intra- and extra-hepatic bile ducts. To date, no animal model has been developed that exhibits all of the attributes of PSC. Rodent models instigated by bacterial cell components or colitis are promising because they may help to explain the strong association between PSC and inflammatory bowel disease (IBD). Other models of direct injury to biliary epithelia, peribiliary vascular endothelia or portal venous endothelia indicate that inflammation, chemokines and cytokines can produce diffuse sclerosis of bile ducts. Models of toxic, infectious or intra-luminal injury of the biliary tract also exhibit focal biliary sclerosis mediated by inflammation and cytokines. The histopathology of several models suggests a sequence of events beginning with secretion of proinflammatory cytokines by activated hepatic macrophages followed by peribiliary infiltration with CD4 and CD8 T cells with a T helper 1 phenotype. These results strongly suggest co-ordinated, pathogenetic roles for both the innate and adaptive immune responses. However, the stimuli that initiate and perpetuate peribiliary fibrosis remain unknown. Interestingly, several models are also associated with the development of anti-neutrophil cytoplasmic antibodies that react in a perinuclear and cytoplasmic pattern similar to that observed in patients with ulcerative colitis and/or PSC. Finally, models of extra-hepatic biliary obstruction continue to provide important information about the pathogenesis of portal fibrosis and secondary biliary cirrhosis that occurs in PSC and other diseases with obstruction of bile flow. Future studies in either existing or new animal models should advance our understanding of the pathogenesis of PSC, the major prerequisite for the development of effective therapies. PMID- 11492971 TI - Immunogenetics in PSC. AB - Primary sclerosing cholangitis (PSC) does not exhibit simple Mendelian inheritance attributable to a single gene locus and our knowledge of the genetics of this complex disease is based entirely on case-control studies of candidate genes. The prime candidates in PSC are inherited variation (polymorphism) in the genes that regulate the immune response, especially the genes of the major histocompatability complex (MHC). Thus far, five different human leukocyte antigen (HLA) haplotypes have been associated with PSC: three with increased risk of disease and two with reduced risk. More recently studies of non-MHC genes have failed to associate PSC with several cytokine genes (IL-1 and IL-10), with FAS (TNFRSF6), with TGFbeta-1, or with CCR-5 but have found genetic links with MMP-3 and disease progression, whilst the potential role of CTLA-4 gene polymorphism remains in question. With the completion of the human genome project, understanding the genetics of complex (non-Mendelian) disease is a major priority for the research community and the studies summarized herein may guide these future investigations. PMID- 11492972 TI - Anti-neutrophil antibodies in primary sclerosing cholangitis. AB - Anti-neutrophil specific antibodies are detected in up to 88% of patients with PSC. The labelling pattern of neutrophils produced by these antibodies when examined by indirect immunofluorescence microscopy is distinct from that produced by anti-neutrophil cytoplasmic antibodies (ANCA) in vasculitic diseases. The antigen(s) recognized by anti-neutrophil antibodies in PSC is not yet known but appears to be localized to the periphery of the nucleus. Accordingly, the term peripheral anti-neutrophil nuclear antibodies (p-ANNA) is more appropriate than the frequently used p-ANCA. As the titre of p-ANNA in PSC does not correlate with disease-specific clinical parameters, they are not useful markers for the management of patients. However, the high prevalence of p-ANNA in PSC makes them a reasonable diagnostic marker if used in conjunction with other standard diagnostic tests. The role of anti-neutrophil antibodies in the pathogenesis of PSC, if any, remains to be established. PMID- 11492973 TI - Hepatobiliary and extra-hepatic malignancies in primary sclerosing cholangitis. AB - The increased risk for cholangiocarcinoma in primary sclerosing cholangitis (PSC) is well established, but the factors responsible for the malignant development in the bile ducts in this disease are not known. The pathogenesis of cholangiocarcinoma in PSC including the role of chronic inflammation and oncogenic mutations will be discussed. Cholangiocarcinoma is a leading cause of death in PSC and the prognosis even after liver transplantation is poor, with a median survival after cancer diagnosis of 5 months. Therefore, it is of great importance to identify PSC patients who are at risk of developing cholangiocarcinoma in order to transplant them before cancer has developed. PMID- 11492974 TI - Medical and endoscopic treatment in primary sclerosing cholangitis. AB - Primary sclerosing cholangitis is an important cause of chronic cholestatic liver disease. The aetiology is still unknown and an immunological basis is discussed. The disease results in diffuse narrowing and irregularities of intra- and extra hepatic bile ducts that may lead to biliary cirrhosis. Progression of the disease is highly variable and fluctuating. An important issue is the risk for developing cholangiocarcinoma. For end-stage disease liver transplantation is the only therapeutic option. If strictures of the extra-hepatic bile ducts are demonstrable, endoscopic interventions are effective palliative treatment options. The use of immunosuppressive or anti-inflammatory drugs has been shown to have no influence on the course of primary sclerosing cholangitis. PMID- 11492975 TI - Liver transplantation for primary sclerosing cholangitis: timing, outcome, impact of inflammatory bowel disease and recurrence of disease. AB - Over the past decade, the outcome of liver transplantation in primary sclerosing cholangitis (PSC) patients with end-stage liver disease has improved significantly with many centres reporting 1-year patient and graft survival of 90 97% and 85-88%, respectively. Based on these results, liver transplantation has emerged as the treatment of choice for PSC patients. Specific complications related to PSC remain problematical. Inflammatory bowel disease (IBD) occurs in 70% of patients, and there is a distinctly increased risk of colorectal neoplasia both pre- and post-transplantation. Furthermore, symptoms related to IBD post transplantation can become severe and lead to the need for proctocolectomy. Cholangiocarcinoma remains a major risk facing the PSC patient and develops in 15 30% of patients. Markers to detect the early neoplastic changes of cholangiocarcinoma are not available. To date, outcome following liver transplantation in PSC patients who have associated cholangiocarcinoma has been dismal. However, those patients who are found to have an incidental cholangiocarcinoma have an acceptable low incidence of recurrence of disease. To assess optimal timing of liver transplantation, natural history risk scores have been developed and utilized. Utilizing such risk scores, estimated survival for the individual PSC patient can be obtained. Finally, there is an increased incidence of both acute and chronic rejection, hepatic artery thrombosis and biliary stricturing in PSC patients undergoing liver transplantation. A late rise in serum alkaline phosphatase level is almost always indicative of biliary stricturing and recurrence of disease. Approximately 20% of patients followed for 5 years or more will have recurrence of PSC documented both on cholangiography and histology. PMID- 11492976 TI - Sclerosing cholangitis in the paediatric patient. AB - Sclerosing cholangitis in childhood is a heterogeneous condition, which has different aetiologies. Sclerosing cholangitis may be inherited and diagnosed in the neonatal period (neonatal sclerosing cholangitis); it may present later with features of autoimmunity (autoimmune sclerosing cholangitis); or it may be associated with a variety of disorders, including Langerhans cell histiocytosis, immunodeficiency, psoriasis, cystic fibrosis, reticulum cell sarcoma and sickle cell anaemia. In contrast to the experience in adult patients, sclerosing cholangitis occurring as an individual disease (primary sclerosing cholangitis) is rare. The initiating events and possible pathogenic mechanisms differ in the various forms of sclerosing cholangitis and are still obscure. Treatment and prognosis depend on the type of sclerosing cholangitis present. PMID- 11492979 TI - Antibodies against oxidized LDL for non-invasive diagnosis of atherosclerotic vascular disease. PMID- 11492980 TI - Endothelin: a new marker of risk of rapid coronary stenosis progression in patients with stable angina? PMID- 11492981 TI - Studying the neuronal side of the synaptic cleft. A tool for investigating the paradox of sympathetic nervous system and heart failure in dilated cardiomyopathy. PMID- 11492982 TI - Why patients with heart failure die. PMID- 11492983 TI - Early atrial defibrillation; "a stitch in time saves nine" or "haste makes waste"? PMID- 11492984 TI - Guidelines for the diagnosis and treatment of chronic heart failure. PMID- 11492985 TI - Platelet activation: assessment and quantification. PMID- 11492986 TI - Autoantibodies against oxidized low density lipoproteins in patients with stable angina, unstable angina or peripheral vascular disease; pathophysiological implications. AB - BACKGROUND: Antibody antioxidized low density lipoproteins (oxLDL) might play a role both in atherogenesis and in the pathogenesis of acute coronary syndromes. METHODS AND RESULTS: Antibody titres to oxLDL and levels of C-reactive protein were compared in unstable angina, stable angina or peripheral artery disease. Antibody titres to LDL oxidated by CuSO(4)for 2, 4 and 18 h (Cu-oxLDL-Ab(2-4-18)) or by peroxidase (HRP-oxLDL-Ab) were assessed by ELISA. Cu-oxLDL-Ab(2-4-18)were consistently higher in peripheral artery disease than in unstable angina (P<0.001, P<0.001, P=0.01, respectively) or in stable angina (P<0.001, P=0.01, P=ns) but similar in unstable and stable angina. Accordingly, HRP-oxLDL-Ab were higher in peripheral artery disease than in unstable angina (P<0.001) or stable angina (P=0.04) but similar in unstable and stable angina. The number of arterial stenoses was higher in peripheral artery disease than unstable and stable angina (P<0.01). Cu-oxLDL-Ab and HRP-oxLDL-Ab correlated with the severity of atherosclerosis (P<0.01, R=0.4;P=0.02, R=0.3 respectively). Conversely, C reactive protein levels were higher in unstable than in stable angina (P<0.001) or in peripheral artery disease (P<0.03) but similar in stable angina and peripheral artery disease and did not correlate with the severity of atherosclerosis. CONCLUSION: The autoimmune response to oxLDL is likely to play an important role in atherogenesis but not in precipitating acute coronary syndromes. PMID- 11492987 TI - Increased plasma endothelin levels in angina patients with rapid coronary artery disease progression. AB - AIMS: To investigate the association between plasma endothelin levels and rapid coronary artery disease progression, as assessed by quantitative angiography. METHODS AND RESULTS: Changes in diameter were assessed in 224 coronary stenoses of 92 consecutive patients (62 men) with chronic stable angina pectoris who were on a waiting list for routine coronary angioplasty and underwent coronary angiography on two occasions: the first (diagnostic) angiogram was carried out at study entry and the second 5.5+/-3.0 months later, immediately prior to coronary angioplasty. A digital quantitative angiographic analysis system was used to assess differences in stenosis diameter between the first and second angiogram. Plasma immunoreactive endothelin levels were estimated by radioimmunoassay at study entry. Rapid coronary artery disease progression occurred in 29 (31.5%) patients according to pre-established criteria: 12 (41%) had a > or =10% diameter reduction of at least one pre-existing stenosis > or =50%, 10 (34%) had a > or =30% diameter reduction of a pre-existing stenosis <50%, 5 (17%) patients developed a new stenosis and 2 (7%) had progression of a lesion to total occlusion by the second angiogram. Baseline demographic, clinical and angiographic data were similar in patients with and without stenosis progression. Plasma endothelin levels were significantly higher in patients with rapid disease progression than in those without (5.7+/-2.0 pg. ml(-1)vs 3.9+/-1.6 pg. ml(-1), P<0.001). Multiple logistic regression analysis revealed that endothelin was an independent predictor of disease progression (P=0.001). Moreover, endothelin levels above 4.26 pg. ml(-1)(the median of the total endothelin concentrations) were associated with a sixfold increase in the risk of developing rapid stenosis progression. CONCLUSIONS: Plasma endothelin is raised in patients with coronary artery disease progression and may be a marker of risk of rapid stenosis progression. Endothelin may also play a pathogenic role in this process. PMID- 11492988 TI - The influence of stent length on clinical and angiographic outcome in patients undergoing elective stenting for native coronary artery lesions; final results of the Magic 5L Study. AB - AIMS: To prospectively evaluate the influence of stent length on 6 month clinical and angiographic outcome, in patients with native coronary lesions up to 45 mm in length, undergoing elective Magic Wallstent implantation. METHODS AND RESULTS: On the basis of pre-procedural angiography, 276 patients (aged 61.3+/-10.2 years; 78.6% male; 41.7% unstable angina) with a total of 302 lesions were prospectively assigned to one of five different length categories of Magic Wallstent. Angiography in multiple matched projections before and after implantation and at 6 months follow-up was analysed at the core laboratory. Primary end-points for the efficacy analysis were cumulative incidence of major adverse cardiac events and quantitative coronary angiography analysis 6 months after stent implantation. Magic Wallstent implantation was successful in 301 of 302 lesions and in 98.6% a residual stenosis <20% by online quantitative coronary angiography was achieved. At 30 days, 6.2% (1.8% subacute occlusion) of patients had experienced major adverse cardiac events, 27.5% at 6 months and 30.4% at 9 months. Angiographic restenosis occurred in 37%. Restenosis rates for the mini, extra-short, short, medium and long Wallstent groups were 25.9%, 25%, 22.6%, 36.2% and 67.5%, respectively. Multivariate analysis revealed stent length to be independently associated with greater angiographic restenosis and major adverse cardiac events. CONCLUSIONS: While shorter Magic Wallstents provided late outcomes comparable with short balloon-expandable stents, excessive restenosis with longer Wallstents should obviate their use in elective percutaneous intervention. Long coronary lesions provide a challenging substrate for emerging antirestenosis therapies, such as stent coatings and brachytherapy. PMID- 11492989 TI - Relationship between altered sympathetic innervation, oxidative metabolism and contractile function in the cardiomyopathic human heart; a non-invasive study using positron emission tomography. AB - AIMS: To identify functional and metabolic correlates of impaired presynaptic sympathetic innervation in the cardiomyopathic human heart using non-invasive correlative imaging. METHODS AND RESULTS: In 10 patients with idiopathic dilated cardiomyopathy, presynaptic catecholamine uptake sites were quantified by positron emission tomography with C-11 hydroxyephedrine. Oxidative metabolism was measured using C-11 acetate. Global and regional function was assessed by tomographic radionuclide angiography. Left ventricular ejection fraction in patients was 19%+/-10%. Myocardial hydroxyephedrine retention was abnormally low in 58%+/-38% of the left ventricles. Globally and regionally, hydroxyephedrine retention was significantly correlated with ventricular function (r=0.67, P=0.03 with left ventricular ejection fraction; r=0.31, P<0.01 with regional endocardial shortening). Multivariate analysis confirmed hydroxyephedrine retention as the closest independent determinant of left ventricular ejection fraction. Oxidative metabolism was determined by rate pressure product as a measure of workload (r=0.78, P<0.01) and peripheral vascular resistance as a measure of afterload (r= 0.61, P=0.06), but did not correlate with hydroxyephedrine retention (r=0.08 for global, r=0.04 for regional parameters). CONCLUSION: Alterations of presynaptic sympathetic innervation in dilated cardiomyopathy are associated with impaired contractile function, suggesting a common pathogenetic pathway. Overall oxidative metabolism, however, was not directly correlated with these findings. Normal regulatory mechanisms for oxidative metabolism were operational. PMID- 11492990 TI - Cardiovascular critical event pathways for the progression of heart failure; a report from the ATLAS study. AB - AIMS: To determine the sequence of critical cardiovascular events in the progression of heart failure, and whether aetiology or high-dose vs low-dose lisinopril affected these pathways. METHODS AND RESULTS: This was a post-hoc investigation of the ATLAS database, which comprised 3164 patients with chronic heart failure, randomized to low- (2.5-5.0 mg. day(-1)) or high-dose (32.5-35.0 mg. day(-1)) lisinopril, followed up for a median of 46 months. Two-thirds (64.3%) of patients had heart failure attributed to ischaemic heart disease. During the study, most patients (61.1%) had at least one cardiovascular hospitalization and 42.5% of all patients died: most deaths (88.2%) were cardiovascular. Nearly half (49.7%) of the cardiovascular deaths were considered sudden and 45.2% of cardiovascular deaths occurred as the first cardiovascular event. A third (30.2%) of deaths resulted from heart failure and were generally preceded by hospitalization, either for heart failure (85.5%), myocardial ischaemic events (21.7%) or arrhythmias (18.0%). Compared with low-dose, high dose lisinopril was associated with a lower risk of death or hospitalization for any reason (P=0.002) and death or hospitalization with worsening heart failure (P<0.001). High-dose lisinopril delayed the time to all-cause mortality and hospitalization for chronic heart failure by 7.1 months. CONCLUSIONS: Vascular and arrhythmic events may not only be important precipitants of sudden death, but were also seen to contribute to the progression of heart failure. A reduction in vascular events, as well as benefits on ventricular remodelling, could account for the decrease in death or hospitalization with high-dose lisinopril. PMID- 11492991 TI - Atrial defibrillation threshold in humans minutes after atrial fibrillation induction; "A stitch in time saves nine". AB - AIMS: To assess the effects of atrial fibrillation duration on the defibrillation threshold in atrial fibrillation patients seconds or minutes after initiation of the arrhythmia. METHODS AND RESULTS: Nineteen patients with recurrent symptomatic atrial fibrillation were evaluated. After programmed induction of atrial fibrillation, the defibrillation threshold was assessed after two sequential periods of arrhythmia in the same patient: an "ultrashort" period of 30 s duration and a "short" period, which lasted 10 min. After the specified period, internal cardioversion was attempted using a balloon-guided catheter that allows the delivery of biphasic shocks between one electrode array placed in the left pulmonary artery and a proximal electrode array on the lateral right atrial wall. The defibrillation threshold was assessed with energy steps of 0.5 J with a starting level of 0.5 J. Mean time from induction to successful defibrillation was 92+/-30 s after the "ultrashort" period of atrial fibrillation and 910+/-86 s after the short period. The defibrillation threshold was significantly greater after 10 min of atrial fibrillation than after 30 s of arrhythmia (2.32+/-0.61 J vs 1.31+/-0.66 J, P<0.001). Clinical data were not found to affect the defibrillation threshold. CONCLUSIONS: Prolongation of atrial fibrillation over minutes in patients with paroxysmal arrhythmia increases the energy requirements for successful defibrillation. PMID- 11492992 TI - Long-term follow-up of children and adolescents with syncope; predictor of syncope recurrence. AB - BACKGROUND: Previous studies have shown that investigation by tilt testing is very appropriate in paediatrics, but the characteristics of children and adolescents who are at high risk of recurrent syncope, once the diagnosis is established, remain unclear. This study was set up to analyse the risk factors attributed to syncope recurrence in paediatric patients. METHODS: One hundred and one children and adolescents aged 7 to 18 years, undergoing a tilt test for recurrent syncope, were studied. They were subsequently followed-up in clinic visits with a final interview at the clinic or by telephone at the end of the follow-up period. RESULTS: A head-up tilt test elicited syncope or pre-syncope in 67 children. The positive responses included vasovagal syncope in 58 patients and psychogenic syncope in nine patients. Gender, age, number of pre-tilt test syncopal episodes or duration of symptoms made no difference to children with positive or negative tilt test results. Following the tilt test, 43 of 67 children with a positive tilt test were treated empirically. No treatment was prescribed for the remaining 24 with a positive test, or for those with a negative tilt test. There were no differences between treated and untreated children with respect to the number of pre-tilt test syncopes, duration of symptoms and duration of follow-up. Follow-up data were available in 97 children. During a mean follow-up of 46+/-28 months, syncope recurred in 31 children (32%). The recurrence rate was similar between positive and negative tilt test groups (22/66 vs 9/31, respectively; P=ns), as well as between treated and untreated children (14/43 vs 8/23, respectively; P=ns). When comparing syncope-free children at follow-up in a univariate analysis, children with recurrent syncope reported a greater number of historical syncopal spells (7+/-8 vs 3+/-3, P=0.01). In addition linear correlation (r=0.6, 95% CI 0.47 to 0.72, P<0.0001) was significant between the number of historical syncope episodes and the risk of recurrent syncope. CONCLUSIONS: These findings suggest that the risk of syncope recurrence for children and adolescents with such a history is not correlated to the tilt test result or prophylactic treatment. The number of historical syncopal spells is, however, predictive. PMID- 11492993 TI - Prognostic value of exercise testing. PMID- 11492994 TI - Dissent from the consensus on the redefinition of myocardial infarction. PMID- 11492997 TI - Structure of histone acetyltransferases. AB - Histone acetyltranferase (HAT) enzymes are the catalytic subunits of multisubunit protein complexes that acetylate specific lysine residues on the N-terminal regions of the histone components of chromatin to promote gene activation. These enzymes, which now include more than 20 members, fall into distinct families that generally have high sequence similarity and related substrate specificity within families, but have divergent sequence and substrate specificity between families. Significant insights into the mode of catalysis and histone substrate binding have been provided by the structure determination of the divergent HAT enzymes Hat1, Gcn5/PCAF and Esa1. A comparison of these structures reveals a structurally conserved central core domain that mediates extensive interactions with the acetyl-coenzyme A cofactor, and structurally divergent N and C-terminal domains. A correlation of these structures with other studies reveals that the core domain plays a particularly important role in histone substrate catalysis and that the N and C-terminal domains play important roles in histone substrate binding. These correlations imply a related mode of catalysis and histone substrate binding by a diverse group of HAT enzymes. PMID- 11492998 TI - Drop-off during ribosome hopping. AB - Ribosomes bypass a 50 nucleotide non-coding segment of mRNA between the two open reading frames of bacteriophage T4 gene 60 in order to synthesize a topoisomerase subunit. While nearly all ribosomes appear to initiate bypassing, only 50 % resume translation in the second open reading frame. Failure to bypass is shown here to be independent of the stop codon at the end of the first open reading frame and to be amplified by mutant variants of tRNA(Gly)(2) known to diminish bypassing efficiency. Unproductive bypassing may result from premature dissociation of peptidyl-tRNAs from ribosomes (drop-off) or resumption of translation at inappropriate sites. Assessment of the influence of factors known to induce drop-off reveals that ribosome recycling factor accounts for a small fraction of unproductive bypassing products, but none of the other known factors appear to play a significant role. Resumption of translation at inappropriate sites appears to be minimal, which suggests that spontaneous release of the peptidyl-tRNA may account for the remaining unproductive bypassing products and may be inherent to the gene 60 bypassing mechanism. PMID- 11492999 TI - Altered directionality in the Cre-LoxP site-specific recombination pathway. AB - The site-specific recombinase Cre must employ control mechanisms to impose directionality on recombination. When two recombination sites (locus of crossing over in phage P1, loxP) are placed as direct repeats on the same DNA molecule, collision between loxP-bound Cre dimers leads to excision of intervening DNA. If two sites are placed as inverted repeats, the intervening segment is flipped around. Cre catalyzes these reactions in the absence of protein co-factors. Current models suggest that directionality is controlled at two steps in the recombination pathway: the juxtaposition of loxP sites and the single-strand transfer reactions within the synaptic complex. Here, we show that in Escherichia coli strain 294-Cre, directionality for recombination is altered when the expression of Cre is increased. This leads to deletion instead of inversion on substrates carrying two loxP sites as inverted repeats. The nucleotide sequence composition of loxP sites remaining in aberrant products indicates that site alignment and/or DNA strand transfer in the in vivo Cre-loxP recombination pathway are not always tightly controlled. PMID- 11493000 TI - Functional expression of a chimeric myosin-containing motor domain of Chara myosin and neck and tail domains of Dictyostelium myosin II. AB - We succeeded in expressing a chimeric myosin that comprises the motor domain of characean algal myosin, (the fastest known motor protein), and the neck and tail domains of Dictyostelium myosin II. Although the chimeric myosin showed an ATPase activity comparable to that of muscle myosin (15 times higher than that of the wild-type Dictyostelium myosin II), the motile activity of the chimera was only 1.3 times higher than that of the wild-type. However, this is the first chimeric myosin that showed motile activity faster than at least one of the parent myosins. It was suggested, therefore, that the motor domain of Chara myosin has the potential for performing fast sliding movement. PMID- 11493001 TI - XAFS study of the high-affinity copper-binding site of human PrP(91-231) and its low-resolution structure in solution. AB - Here, we describe the structure of a C-terminal high-affinity copper-binding site within a truncated recombinant human PrP containing residues 91-231, which lacks the octapeptide repeat region. We show that at least two extra co-ordinating groups are involved in binding this copper(II) ion in conjunction with histidine residues 96 and 111 in a region of the molecule known to be critical in conferring strain type. In addition, using X-ray solution scattering, a low resolution shape of PrP(91-231) is provided. The restored molecular envelope is consistent with the picture where the N-terminal segment, residues 91-120, extends out from the previously known globular domain containing residues 121 231. PMID- 11493002 TI - Structure and dimerization of HIV-1 kissing loop aptamers. AB - Dimerization of two homologous strands of genomic RNA is an essential feature of the retroviral replication cycle. In HIV-1, genomic RNA dimerization is facilitated by a conserved stem-loop structure located near the 5' end of the viral RNA called the dimerization initiation site (DIS). The DIS loop is comprised of nine nucleotides, six of which define an autocomplementary sequence flanked by three conserved purine residues. Base- pairing between the loop sequences of two copies of genomic RNA is necessary for efficient dimerization. We previously used in vitro evolution to investigate a possible structural basis for the marked sequence conservation of the DIS loop. In this study, chemical structure probing, measurements of the apparent dissociation constants, and computer structure analysis of dimerization-competent aptamers were used to analyze the dimers' structure and binding. The selected aptamers were variants of the naturally occurring A and B subtypes. The data suggest that a sheared base pair closing the loop of the DIS is important for dimerization in both subtypes. On the other hand, the open or closed state of the last base-pair in the stem differed in the two subtypes. This base-pair appeared closed in the subtype A DIS dimer and open in subtype B. Finally, evidence for a cross-talk between nucleotides 2, 5, and 6 was found in some, but not all, loop contexts, indicating some structural plasticity depending on loop sequence. Discriminating between the general rules governing dimer formation and the particular characteristics of individual DIS aptamers helps to explain the affinity and specificity of loop loop interactions and could provide the basis for development of drugs targeted against the dimerization step during retroviral replication. PMID- 11493003 TI - High-affinity DNA binding of HU protein from the hyperthermophile Thermotoga maritima. AB - Prokaryotic genomes are compacted by association with small basic proteins, generating what has been termed bacterial chromatin. The ubiquitous DNA-binding protein HU serves this function. DNA-binding properties of HU from the hyperthermophilic eubacterium Thermotoga maritima are shown here to differ significantly from those characteristic of previously described HU homologs. Electrophoretic mobility shift analyses show that T. maritima HU (TmHU) binds double-stranded DNA with high affinity (K(d)=5.6(+/-0.7) nM for 37 bp DNA). Equivalent affinity is observed between 4 degrees C and 45 degrees C. TmHU has higher affinity for DNA containing a set of 4 nt loops separated by 9 bp (K(d)=1.4(+/-0.3) nM), consistent with its introduction of two DNA kinks. Using DNA probes of varying length, the optimal binding site for TmHU is estimated at 37 bp, in sharp contrast to the 9-10 bp binding site reported for other HU homologs. Alignment of >60 HU sequences demonstrates significant sequence conservation: A DNA-intercalating proline residue is almost universally conserved, and it is preceded by arginine and asparagine in most sequences, generating a highly conserved RNP motif; V substitutes for R only in HU from Thermotoga, Thermus and Deinococcus. A fivefold increase in DNA-binding affinity is observed for TmHU in which V is replaced with R (TmHU-V61R; K(d)=1.1(+/-0.2) nM), but a change in the trajectory of DNA flanking the sites of DNA intercalation is inferred from analysis of TmHU-V61R binding to DNA modified with 4 nt loops or with substitutions of 5-hydroxymethyluracil for thymine. Survival in extreme environments places unique demands on protection of genomic DNA from thermal destabilization and on access of DNA to the cellular machinery, demands that may be fulfilled by the specific DNA-binding properties of HU and by the fine structure of the bacterial chromatin. PMID- 11493004 TI - DNA cleavage reactions by type II restriction enzymes that require two copies of their recognition sites. AB - Several type II restriction endonucleases interact with two copies of their target sequence before they cleave DNA. Three such enzymes, NgoMIV, Cfr10I and NaeI, were tested on plasmids with one or two copies of their recognition sites, and on catenanes containing two interlinked rings of DNA with one site in each ring. The enzymes showed distinct patterns of behaviour. NgoMIV and NaeI cleaved the plasmid with two sites faster than that with one site and the catenanes at an intermediate rate, while Cfr10I gave similar steady-state rates on all three substrates. Both Cfr10I and NgoMIV converted the majority of the substrates with two sites directly to the products cut at both sites, while NaeI cleaved just one site at a time. All three enzymes thus synapse two DNA sites through three dimensional space before cleaving DNA. With Cfr10I and NgoMIV, both sites are cleaved in one turnover, in a manner consistent with their tetrameric structures, while the cleavage of a single site by NaeI indicates that the second site acts not as a substrate but as an activator, as reported previously. The complexes spanning two sites have longer lifetimes on catenanes with one site in each ring than on circular DNA with two sites, which indicates that the catenanes have more freedom for site juxtaposition than plasmids with sites in cis. PMID- 11493005 TI - Analysis of DNA looping interactions by type II restriction enzymes that require two copies of their recognition sites. AB - Before cleaving DNA substrates with two recognition sites, the Cfr10I, NgoMIV, NaeI and SfiI restriction endonucleases bridge the two sites through 3D space, looping out the intervening DNA. To characterise their looping interactions, the enzymes were added to plasmids with two recognition sites interspersed with two res sites for site-specific recombination by Tn21 resolvase, in buffers that contained either EDTA or CaCl2 so as to preclude DNA cleavage by the endonuclease; the extent to which the res sites were sequestered into separate loops was evaluated from the degree of inhibition of resolvase. With Cfr10I, a looped complex was detected in the presence but not in the absence of Ca(2+); it had a lifetime of about 90 seconds. Neither NgoMIV nor NaeI gave looped complexes of sufficient stability to be detected by this method. In contrast, SfiI with Ca(2+) produced a looped complex that survived for more than seven hours, whereas its looping interaction in EDTA lasts for about four minutes. When resolvase was added to a SfiI binding reaction in EDTA followed immediately by CaCl2, the looped DNA was blocked from recombination while the unlooped DNA underwent recombination. By measuring the distribution between looped and unlooped DNA at various SfiI concentrations, and by fitting the data to a model for DNA binding by a tetrameric protein to two sites in cis, an equilibrium constant for the looping interaction was determined. The equilibrium constant was essentially independent of the length of DNA between the SfiI sites. PMID- 11493006 TI - Mapping of binding sites for nidogens, fibulin-2, fibronectin and heparin to different IG modules of perlecan. AB - Perlecan, a major basement membrane proteoglycan, has a complex modular structure designed for the binding of many cellular and extracellular ligands. Its domain IV, which consists of a tandem of immunoglobulin-like modules (IG2-IG15), is rich in such binding sites, which have been mapped to different modules obtained by recombinant production. Heparin/sulfatide binding was restricted to IG5 and shown to depend on four arginine residues that are close in space in beta strands B and E of the C-type IG fold. The nidogen-1 and nidogen-2 isoforms bind to IG3 with high affinity (K(d) approximately 10 nM). This interaction depends on the globular nidogen domain G2 and is crucial for the formation of ternary complexes with laminins. Two loops of IG3 located between beta strands B/C and F/G, which are spatially close, make a major contribution to binding. Fibronectin binding was localized to IG4-5 and fibulin-2 binds to IG2 and IG13-15 with different affinities. This implicates a complex cluster of heterotypic interaction sites apparently important for the supramolecular organization of perlecan in tissues. PMID- 11493008 TI - Inhibition of human alpha-thrombin by a phosphonate tripeptide proceeds via a metastable pentacoordinated phosphorus intermediate. AB - X-ray crystallographic studies of human alpha-thrombin with a novel synthetic inhibitor, an acyl (alpha-aminoalkyl)phosphonate, reveal the existence of a pentacovalent phosphorus intermediate state. Crystal structures of the complex of alpha-thrombin with the phosphonate compound were determined independently using crystals of different ages. The first structure, solved from a crystal less than seven days old, showed a pentacoordinated phosphorus moiety. The second structure, determined from a crystal that was 12 weeks old, showed a tetracoordinated phosphorus moiety. In the first structure, a water molecule, made nucleophilic by coordination to His57 of alpha-thrombin, is bonded to the pentacoordinated phosphorus atom. Its position is approximately equivalent to that occupied by the water molecule responsible for hydrolytic deacylation during normal hydrolysis. The pentacoordinated phosphorus adduct collapses to give the expected pseudo tetrahedral complex, where the phosphorus atom is covalently bonded to Ser195 O(gamma). The crystallographic data presented here therefore suggest that the covalent bond formed between the inhibitor's phosphorus atom and O(gamma) of Ser195 proceeds via an addition-elimination mechanism, which involves the formation of a pentacoordinate intermediate. PMID- 11493007 TI - Crystal structure of the 20 S proteasome:TMC-95A complex: a non-covalent proteasome inhibitor. AB - The 20 S proteasome core particle (CP), a multicatalytic protease, is involved in a variety of biologically important processes, including immune response, cell cycle control, metabolic adaptation, stress response and cell differentiation. Therefore, selective inhibition of the CP will be one possible way to influence these essential pathways. Recently, a new class of specific proteasome inhibitors, TMC-95s, was investigated and we now present a biochemical and crystallographic characterisation of the yeast proteasome core particle in complex with the natural product TMC-95A. This unusual heterocyclic compound specifically blocks the active sites of CPs non-covalently, without modifying the nucleophilic Thr1 residue. The inhibitor is bound to the CP by specific hydrogen bonds with the main-chain atoms of the protein. Analysis of the crystal structure of the complex has revealed which portions of TMC-95s are essential for binding to the proteasome. This will form the basis for the development of synthetic selective proteasome inhibitors as promising candidates for anti-tumoral or anti inflammatory drugs. PMID- 11493009 TI - Crystal structure of PTP-SL/PTPBR7 catalytic domain: implications for MAP kinase regulation. AB - Protein tyrosine phosphatases PTP-SL and PTPBR7 are isoforms belonging to cytosolic membrane-associated and to receptor-like PTPs (RPTPs), respectively. They represent a new family of PTPs with a major role in activation and translocation of MAP kinases. Specifically, the complex formation between PTP-SL and ERK2 involves an unusual interaction leading to the phosphorylation of PTP-SL by ERK2 at Thr253 and the inactivating dephosphorylation of ERK2 by PTP-SL. This interaction is strictly dependent upon a kinase interaction motif (KIM) (residues 224-239) situated at the N terminus of the PTP-SL catalytic domain. We report the first crystal structure of the catalytic domain for a member of this family (PTP SL, residues 254-549, identical with residues 361-656 of PTPBR7), providing an example of an RPTP with single cytoplasmic domain, which is monomeric, having an unhindered catalytic site. In addition to the characteristic PTP-core structure, PTP-SL has an N-terminal helix, possibly orienting the KIM motif upon interaction with the target ERK2. An unusual residue in the catalytically important WPD loop promotes formation of a hydrophobically and electrostatically stabilised clamp. This could induce increased rigidity to the WPD loop and therefore reduced catalytic activity, in agreement with our kinetic measurements. A docking model based on the PTP-SL structure suggests that, in the complex with ERK2, the phosphorylation of PTP-SL should be accomplished first. The subsequent dephosphorylation of ERK2 seems to be possible only if a conformational rearrangement of the two interacting partners takes place. PMID- 11493010 TI - High resolution crystal structures of T4 phage beta-glucosyltransferase: induced fit and effect of substrate and metal binding. AB - beta-Glucosyltransferase (BGT) is a DNA-modifying enzyme encoded by bacteriophage T4 that transfers glucose from uridine diphosphoglucose to 5-hydroxymethyl cytosine bases of phage T4 DNA. We report six X-ray structures of the substrate free and the UDP-bound enzyme. Four also contain metal ions which activate the enzyme, including Mg(2+) in forms 1 and 2 and Mn(2+) or Ca(2+). The substrate free BGT structure differs by a domain movement from one previously determined in another space group. Further domain movements are seen in the complex with UDP and the four UDP-metal complexes. Mg(2+), Mn(2+) and Ca(2+) bind near the beta phosphate of the nucleotide, but they occupy slightly different positions and have different ligands depending on the metal and the crystal form. Whilst the metal site observed in these complexes with the product UDP is not compatible with a role in activating glucose transfer, it approximates the position of the positive charge in the oxocarbonium ion thought to form on the glucose moiety of the substrate during catalysis. PMID- 11493011 TI - Solution structures by 1H NMR of the novel cyclic trypsin inhibitor SFTI-1 from sunflower seeds and an acyclic permutant. AB - SFTI-1 is a recently discovered cyclic peptide trypsin inhibitor from sunflower seeds comprising 14 amino acid residues. It is the most potent known Bowman-Birk inhibitor and the only naturally occurring cyclic one. The solution structure of SFTI-1 has been determined by 1H-NMR spectroscopy and compared with a synthetic acyclic permutant. The solution structures of both are remarkably similar. The lowest energy structures from each family of 20 structures of cyclic and acyclic SFTI-1 have an rmsd over the backbone and heavy atoms of 0.29 A and 0.66 A, respectively. The structures consist of two short antiparallel beta-strands joined by an extended loop containing the active site at one end. Cyclic SFTI-1 also has a hairpin turn completing the cycle. Both molecules contain particularly stable arrangements of cross-linking hydrogen bonds between the beta-strands and a single disulfide bridge, making them rigid and well defined in solution. These stable arrangements allow both the cyclic and acyclic variants of SFTI-1 to inhibit trypsin with very high potencies (0.5 nM and 12.1 nM, respectively). The cyclic nature of SFTI-1 appears to have evolved to provide higher trypsin inhibition as well as higher stability. The solution structures are similar to the crystal structure of the cyclic inhibitor in complex with trypsin. The lack of a major conformational change upon binding suggests that the structure of SFTI 1 is rigid and already pre-organized for maximal binding due to minimization of entropic losses compared to a more flexible ligand. These properties make SFTI-1 an ideal platform for the design of small peptidic pharmaceuticals or pesticides. PMID- 11493012 TI - Solution structure and function of a conserved protein SP14.3 encoded by an essential Streptococcus pneumoniae gene. AB - Streptococcus pneumoniae is a major human pathogen that causes high mortality and morbidity rates and has developed resistance to many antibiotics. The genome of S. pneumoniae has recently been completely sequenced revealing many genes encoding hypothetical proteins of unknown function. We have found that the gene encoding one such conserved protein, SP14.3, is essential for growth of S. pneumonia. Since it is essential, SP14.3 represents a potential target for drug discovery. Here, we describe the three-dimensional solution structure of SP14.3 as determined by NMR spectroscopy. The structure consists of two domains each with an alpha/beta-fold. The N-terminal domain contains two alpha-helices and a three-stranded beta-sheet, while the C-terminal domain is composed of one alpha helix and a five-stranded beta-sheet. The N-terminal domain of the protein contains a highly negatively charged surface and resembles the fold of the N terminal domain of Thermus thermophilus ribosomal protein S3. The C-terminal domain has a protein fold similar to human small nuclear ribonucleoprotein Sm D3 and Haloarcula marismortui ribosomal protein L21E. The two domains of the protein tumble in solution overall as a whole with an overall molecular rotational correlation time (tau(m)) of 12.9 ns at 25 degrees C. The relative orientation of the two domains is not defined by the nuclear Overhauser effect data. Indeed, residual dipolar couplings and the structure calculations indicate that the relative orientation of the two domains is not rigidly oriented with respect to one another in solution. PMID- 11493013 TI - Microsecond exchange of internal water molecules in bacteriorhodopsin. AB - The proton-conducting pathway of bacteriorhodopsin (BR) contains at least nine internal water molecules that are thought to be key players in the proton translocation mechanism. Here, we report the results of a multinuclear (1H, 2H, 17O) magnetic relaxation dispersion (MRD) study with the primary goal of determining the rate of exchange of these internal water molecules with bulk water. This rate is of interest in current attempts to elucidate the molecular details of the proton translocation mechanism. The relevance of water exchange kinetics is underscored by recent crystallographic findings of substantial variations in the number and locations of internal water molecules during the photocycle. Moreover, internal water exchange is believed to be governed by conformational fluctuations in the protein and can therefore provide information about the thermal accessibility of functionally important conformational substates. The present 2H and 17O MRD data show that at least seven water molecules, or more if they are orientationally disordered, in BR have residence times (inverse exchange rate constant) in the range 0.1-10 micros at 277 K. At least five of these water molecules have residence times in the more restrictive range 0.1-0.5 micros. These results show that most or all of the deeply buried water molecules in BR exchange on a time-scale that is short compared to the rate limiting step in the photocycle. The MRD measurements were performed on BR solubilized in micelles of octyl glucoside. From the MRD data, the rotational correlation time of detergent-solubilized BR was determined to 35 ns at 300 K, consistent with a monomeric protein in complex with about 150 detergent molecules. The solubilized protein was found to be stable in the dark for at least eight months at 277 K. PMID- 11493014 TI - On Setting Response Criteria for Calibrated Subjective Probability Estimates. AB - Framing the issue of subjective probability calibration in signal-detection theory terms, this paper first proves a theorem regarding the placement of well calibrated response criteria and then develops an algorithm guaranteed to find such criteria, should they exist. Application of this algorithm to tasks varying in difficulty and number of response categories shows that perfect calibration is easiest to attain under median difficulty levels (d' approximately 1.4) and is practically or theoretically impossible to attain when the task is either very hard (d' approximately 0.5) or very easy (d' approximately 10). Implications for calibration research, including the hard-easy effect, are discussed. Copyright 2001 Academic Press. PMID- 11493015 TI - Judgments of Frequency and Recency in a Distributed Memory Model. AB - TODAM is a theory of distributed associative memory based on the convolution correlation formalism of A. Borsellino and T. Poggio (1973, Kybernetik, 122, 113 122), and TODAM2 is a revised version which includes context, auto-associations for binding, a dual basis for item information, and mediators for associative information. It can explain some complex interactions (differential forgetting and differential attention) between item and associative information. In this paper we derive the basic expressions for the memory-probe dot product for the 2x2 cases (item and pair study crossed with item and pair probe), and then we apply these expressions to judgments of frequency (JOF) and judgments of recency (JOR). We report an experiment which tests both JOF and JOR for single items with post-cuing to control for encoding strategies and suggest an attenuation factor for repetition to improve the fits. With attenuation, TODAM2 can fit the JOR data, but the JOF fits, while not too bad, consistently predict too much dependence between item and associative information. Copyright 2001 Academic Press. PMID- 11493016 TI - Extending General Processing Tree Models to Analyze Reaction Time Experiments. AB - General processing tree (GPT) models are usually used to analyze categorical data collected in psychological experiments. Such models assume functional relations between probabilities of the observed behavior categories and the unobservable choice probabilities involved in a cognitive task. This paper extends GPT models for categorical data to the analysis of continuous data in a class of response time (RT) experiments in cognitive psychology. Suppose that a cognitive task involves several discrete processing stages and both accuracy (categorical) and latency (continuous) measures are obtained for each of the response categories. Furthermore, suppose that the task can be modeled by a GPT model that assumes serialization among the stages. The observed latencies of the response categories are functions of the choice probabilities and processing times (PT) at each of the processing stages. The functional relations are determined by the processing structure of the task. A general framework is presented and it is applied to a set of data obtained from a source monitoring experiment. Copyright 2001 Academic Press. PMID- 11493017 TI - A Mathematical Model of Depth Displacement with a Contracting Bar. AB - When a vertical line segment contracts at both ends according to a (decelerating) time law of hyperbolic type, it appears to rotate around its midpoint. The phenomenon is quite surprising from the point of view of projective geometry as the segment should rather appear to recede along the sagittal plane. Apparent displacement in depth is however obtained when the bar simultaneously contracts and is displaced laterally on the frontal plane. But, here again, projective expectations are contradicted, because apparent displacement in depth occurs whether movement of the bar in two dimensions is hyperbolic (decelerating), uniform, harmonic (accelerating), or a mixture of the three. It is suggested here that the visual system operates in such a way as to minimize the differences between the lengths of the velocity vectors of all points of a moving configuration. The mathematical model derived from this hypothesis allows qualitative and quantitative predictions that are in good agreement with experimental results. Copyright 2001 Academic Press. PMID- 11493018 TI - Strategic Polarization. AB - In joint decision making, similarly minded people may take opposite positions. Consider the example of a marriage in which one spouse gives generously to charity while the other donates nothing. Such "polarization" may misrepresent what is, in actuality, a small discrepancy in preferences. It may be that the donating spouse would like to see 10% of their combined income go to charity each year, while the apparently frugal spouse would like to see 8% donated. A simple game-theoretic analysis suggests that the spouses will end up donating 10% and 0%, respectively. By generalizing this argument to a larger class of games, we provide strategic justification for polarization in many situations such as debates, shared living accommodations, and disciplining children. In some of these examples, an arbitrarily small disagreement in preferences leads to an arbitrarily large loss in utility for all participants. Such small disagreements may also destabilize what, from game-theoretic point of view, is a very stable equilibrium. Copyright 2001 Academic Press. PMID- 11493019 TI - TELEGRAPHIC REVIEWS. PMID- 11493020 TI - Insulin-like growth factor-I in muscle metabolism and myotherapies. AB - The critical anabolic and trophic role of signaling by insulin-like growth factors (IGF) I and II via the type-I IGF receptor (IGF-IR) is reviewed throughout the life of skeletal myocytes. The proliferative effects of IGF-IR stimulation, both during embryogenesis and during satellite cell proliferation following denervation or muscle injury, are mediated primarily through activation of mitogen-activated protein kinases. Signaling through phosphatidylinositol 3 kinase is essential to muscle protein synthesis and glucose uptake and may contribute to the observed resilience of mature muscle to programmed cell death. Degeneration or inhibition of the GH--IGF-I axis by aging, cachexia, sepsis, diabetes, drugs, and disuse all enhance muscle catabolism, and opposition of these effects by IGF-I may form the basis of effective myotherapy. PMID- 11493021 TI - Permeability of proteins at the blood-brain barrier in the normal adult mouse and double transgenic mouse model of Alzheimer's disease. AB - The permeability of albumin, insulin, and human A beta 1--40 at the blood-brain barrier (BBB) was determined in the normal adult mouse (B6/SJL) and in the double transgenic Alzheimer mouse (APP, PS1) by using an I.V. bolus injection technique to quantify the permeability coefficient-surface area (PS) product for each protein after correction for the residual plasma volume (V(p)) occupied by the protein in the blood vessels of different brain regions using a second aliquot of the same protein radiolabeled with a different isotope of iodine ((125)I vs (131)I) as a vascular space marker. This technology for quantifying BBB permeability of proteins was adapted from the rat to the mouse and involved catheterizing the femoral artery and vein of the mouse instead of the brachial artery and vein as for the rat. Because of the smaller blood volume in the mouse, serial sampling (20 microl) of blood from the femoral artery of the mouse was performed and directly TCA precipitated to generate a whole blood washout curve for the intact protein. When similar blood sampling techniques were used in the rat, the PS values for albumin and insulin at the BBB were similar in these two species. In the double transgenic mouse, the V(p) values for albumin were significantly increased 1.4- to 1.6-fold in five of six brain regions compared to the normal adult mouse, which indicated increased adherence of albumin to vessel walls. As a result, the PS values were significantly decreased, from 1.4- to 3.2 fold, which likely reflected decreased transport of albumin by passive diffusion. In contrast, insulin, which is taken up into the brain by a receptor-mediated transport mechanism at the BBB, showed no significant difference in the V(p) values but a significant increase in the PS values in four of six brain regions. This suggests a compensatory mechanism in the Alzheimer's transgenic brain whereby there is an increased permeability to insulin at the BBB. Surprisingly, there was no significant difference in the V(p) or PS values for human A beta 1- 40 at the BBB in the double transgenic Alzheimer mouse at 24, 32, or 52 weeks of age, when there is both significant A beta levels in the plasma and amyloid burden in the brains of these animals. These data suggest that there is not an alteration in permeability to human A beta 1--40 at the BBB with increasing amyloid burden in the double transgenic Alzheimer mouse. Although these observations suggest structural alterations at the BBB, they do not support the concept of extensive BBB damage with substantial increases in BBB permeability in Alzheimer's disease. PMID- 11493023 TI - Regionalized neurofilament accumulation and motoneuron degeneration are linked phenotypes in wobbler neuromuscular disease. AB - Abnormal neurofilament aggregates are pathological hall-mark of most neurodegenerative diseases, although their pathogenic role remains unclear. Increased expression of medium neurofilament (NFM) is an early molecular marker of wobbler mouse, an animal model of motoneuron disease. In the wr/wr, a vacuolar neuronal degeneration (VND) starts at 15 days postnatally, selectively in cervical spinal cord and brain stem motoneurons. Here we show that nfm gene hyperexpression is restricted to the aforementioned motoneurons and is specific for wr mutation. NF proteins accumulate in wr/wr before VND. wr/+ mice, which are asymptomatic, show intermediate NF accumulation between wr/wr and +/+ littermates, suggesting a gene dosage dependence of the wobbler pathology. Altogether our data indicate that NF hyperexpression and regionalized motoneuron degeneration are linked to the wr mutation, although with a still unknown relationship to the mutant gene activity. PMID- 11493022 TI - Cleavage of bid may amplify caspase-8-induced neuronal death following focally evoked limbic seizures. AB - The mechanism by which seizures induce neuronal death is not completely understood. Caspase-8 is a key initiator of apoptosis via extrinsic, death receptor-mediated pathways; we therefore investigated its role in mediating seizure-induced neuronal death evoked by unilateral kainic acid injection into the amygdala of the rat, terminated after 40 min by diazepam. We demonstrate that cleaved (p18) caspase-8 was detectable immediately following seizure termination coincident with an increase in cleavage of the substrate Ile-Glu-Thr-Asp (IETD)-p nitroanilide and the appearance of cleaved (p15) Bid. Expression of Fas and FADD, components of death receptor signaling, was increased following seizures. In vivo intracerebroventricular z-IETD-fluoromethyl ketone administration significantly reduced seizure-induced activities of caspases 8, 9, and 3 as well as reducing Bid and caspase-9 cleavage, cytochrome c release, DNA fragmentation, and neuronal death. These data suggest that intervention in caspase-8 and/or death receptor signaling may confer protection on the brain from the injurious effects of seizures. PMID- 11493024 TI - Tumor necrosis factor-alpha induces cFOS and strongly potentiates glutamate mediated cell death in the rat spinal cord. AB - Excitotoxic cell death due to glutamate release is important in the secondary injury following CNS trauma or ischemia. Proinflammatory cytokines also play a role. Both glutamate and tumor necrosis factor-alpha (TNF(alpha)) are released immediately after spinal cord injury. Neurophysiological studies show that TNF(alpha) can potentiate the effects of glutamatergic afferent input to produce hyperactivation of brain-stem sensory neurons. Therefore, we hypothesized that TNF(alpha) might act cooperatively with glutamate to affect cell death in the spinal cord as well. Nanoinjections of either TNF(alpha) (60 pg) or kainate (KA; 32 ng) alone into the thoracic gray resulted in almost no tissue damage or cell death 90 min after injection. However, the combination of TNF(alpha) plus KA at these same doses produced a large area of tissue necrosis and neuronal cell death, an effect which was blocked by the AMPA receptor antagonist CNQX (17 ng). These results suggest that secondary injury may involve potentiation of AMPA receptor-mediated excitatory cell death by TNF(alpha). PMID- 11493025 TI - Retrovirus-mediated gene transfer and galactocerebrosidase uptake into twitcher glial cells results in appropriate localization and phenotype correction. AB - Galactocerebrosidase (GALC) is deficient in all tissues from human patients and animal models with globoid cell leukodystrophy (GLD) or Krabbe disease. The deficiency results in decreased lysosomal catabolism of certain galactolipids including galactosylceramide and psychosine that are synthesized maximally during myelination. According to current theories, the accumulation of psychosine in humans and animals with GLD induces oligodendrocyte degeneration and myelination ceases. Transduction of oligodendrocytes from twitcher mice with a retroviral vector containing the GALC cDNA can correct the enzyme deficiency in these cells. Our data show that twitcher astrocytes and oligodendrocytes can internalize exogenous GALC, as well as donate the enzyme to the mutant glial cells. Antibodies against human GALC localized the GALC antigen in retrovirally transduced cells and cells receiving enzyme via cell to cell secretion and uptake to the lysosomal fraction. In fact immunocytochemical studies in transduced oligodendrocytes revealed that the GALC colocalizes in vesicles lysosomal associated membrane protein-2 (LAMP2) (+). Moreover, labeling cells with anti GALC and a marker for oligodendrocytes demonstrated that, upon differentiation, transduced, twitcher oligodendrocytes attained the normal branched process configuration, while untransduced cells show only abnormal morphology. Phenotype correction in mutant oligodendrocytes has also been observed after enzyme transfer. These studies indicate that GALC activity supplied to cultured oligodendrocytes from twitcher mice by different methods can correct the pathological phenotype of these cells. PMID- 11493026 TI - Impaired neuronal plasticity in transgenic mice expressing human apolipoprotein E4 compared to E3 in a model of entorhinal cortex lesion. AB - The apolipoprotein E (APOE) epsilon 4 allele is a major risk factor for late onset familial and sporadic Alzheimer's disease (AD) and is associated with a poor outcome after brain injury. Each apoE isoform is suggested to have differential effects on neuronal repair mechanisms within the CNS. In the present study, APOE genotype influence on the immediate response to injury and subsequent repair process was examined in a line of transgenic APOE mice possessing human APOE gene insertions (epsilon 3 and epsilon 4). Quantification of synaptophysin and GAP-43 immunoreactivity was used to measure the extent of degeneration and regeneration after entorhinal cortex lesion (ECL). Progressive neurodegenerative decline occurred in the ipsilateral dentate gyrus until day 28 post-ECL which was more severe in APOE epsilon 3 mice compared to APOE epsilon 4 mice. By day 90 post-ECL compensatory sprouting and reactive synaptogenesis had taken place in the dentate gyrus of APOE epsilon 3 mice such that GAP-43 and synaptophysin immunoreactivity had returned to prelesion levels. In contrast, APOE epsilon 4 mice displayed significant deficits in synaptophysin and GAP-43 immunostaining compared to the APOE epsilon 3 mice (P < 0.05). Expansion of the inner molecular layer (IML) was used as a measure of the sprouting index from the commissural associational pathway and by day 90 post-ECL the IML width in APOE epsilon 3 mice had increased by 45% but only 20% in APOE epsilon 4 mice (P < 0.0001). ApoE immunoreactivity was increased within the neuropil and glia to the same extent in APOE epsilon 3 and APOE epsilon 4 mice post-ECL. There was no significant difference in the deposition and clearance of degeneration products between APOE epsilon 3 and epsilon 4 mice post-ECL. These results indicate that neuronal plasticity is impaired in transgenic mice possessing human APOE epsilon 4 alleles compared to APOE epsilon 3. These isoform-specific differences in plasticity may relate to the severity of AD and poor, long-term recovery after head injury in APOE epsilon 4 individuals. PMID- 11493027 TI - Pyridoxine-induced neuropathy in rats: a sensory neuropathy that responds to 4 methylcatechol. AB - Sensory neuropathies are frequently associated with diabetes or with antimitotic treatments in humans suffering from cancer, and are in this case the most important limitation to the use of antimitotic drugs. For this reason, there is a need to establish and validate animal models of sensory neuropathies that could be routinely used, together with the already known models, for studying and evaluating the effects of putative neuroprotective compounds. In the present study, we prove by behavioral and electromyographical analyses that (a) it is possible to induce a nonlethal, exclusively sensory, reversible neuropathy by intoxicating rats with large amounts of pyridoxine, using a new schedule of intoxication; (b) 4-methylcatechol, a drug known to induce nerve growth factor synthesis, improves the clinical status of pyridoxine-intoxicated animals, shortens the duration of the disease, and restores the morphological integrity of the sensory fibers. Owing to its mode of installation and its clinical features, we propose that this model be used as an additional model for preclinical studies of neuroprotective drugs. PMID- 11493028 TI - Involvement of GDNF in neuronal protection against 6-OHDA-induced parkinsonism following intracerebral transplantation of fetal kidney tissues in adult rats. AB - Exogenous application of transforming growth factors-beta (TGF beta) family proteins, including glial cell line-derived neurotrophic factor (GDNF), neurturin, activin, and bone morphogenetic proteins, has been shown to protect neurons in many models of neurological disorders. Finding a tissue source containing a variety of these proteins may promote optimal beneficial effects for treatment of neurodegenerative diseases. Because fetal kidneys express many TGF beta trophic factors, we transplanted these tissues directly into the substantia nigra after a unilateral 6-hydroxydopamine lesion. We found that animals that received fetal kidney tissue grafts exhibited (1) significantly reduced hemiparkinsonian asymmetrical behaviors, (2) a near normal tyrosine hydroxylase immunoreactivity in the lesioned nigra and striatum, (3) a preservation of K(+) induced dopamine release in the lesioned striatum, and (4) high levels of GDNF protein within the grafts. In contrast, lesioned animals that received grafts of adult kidney tissues displayed significant behavioral deficits, dopaminergic depletion, reduced K(+)-mediated striatal dopamine release, and low levels of GDNF protein within the grafts. The present study suggests that fetal kidney tissue grafts can protect the nigrostriatal dopaminergic system against a neurotoxin-induced parkinsonism, possibly through the synergistic release of GDNF and several other neurotrophic factors. PMID- 11493029 TI - Increased cystatin C in astrocytes of transgenic mice expressing the K670N-M671L mutation of the amyloid precursor protein and deposition in brain amyloid plaques. AB - Cystatin C is an essential secretory cofactor for neurogenesis with potent protease inhibitor activities. Polymorphisms of cystatin C are genetically associated with Alzheimer's disease (AD), and the L68Q mutation causes hereditary cerebral hemorrhage with amyloidosis of the Icelandic type, in which cystatin C and beta-amyloid are colocalized in cortical blood vessels. To determine whether cystatin C and beta-amyloid also colocalize in brain amyloid plaques, we analyzed transgenic mice expressing the Swedish APP (SweAPP) mutation. We found high levels of cystatin C in astrocytes surrounding beta-amyloid plaques, and discrete layers of cystatin C attached to amyloid plaque cores covered by a layer of beta amyloid. In addition, cystatin C accumulated in reactive astrocytes throughout the brain, independently of, and before the onset of, amyloid plaque formation. These results show that expression of SweAPP is associated with increased cystatin C in reactive astrocytes, and they suggest an early role of cystatin C in appositional amyloid plaque growth. PMID- 11493030 TI - Adenovirus-mediated GDNF and CNTF pretreatment protects against striatal injury following transient middle cerebral artery occlusion in mice. AB - During the last few years, adenoviral gene transfer techniques have achieved increasing interest in the treatment of neurodegenerative diseases. However, gene therapy requires that delivered genes are translated into proteins. This may pose a problem in focal ischemia where protein synthesis is compromized. The present study was conducted to find out the feasibility of adenoviral GDNF and CNTF delivery in transient focal ischemia, as induced by 30 min of intraluminar middle cerebral artery (MCA) occlusion in mice. Injections of vehicle, of an adenoviral vector deleted in the E1 region (Ad-dE1) and of vectors expressing the GDNF (Ad GDNF), CNTF (Ad-CNTF), or GFP (Ad-EGFP) gene from a CMV promoter were stereotactically placed in the dorsolateral striatum, i.e., the core of the MCA territory, and focal ischemia was induced seven days later. Thread occlusion resulted in disseminated injury of the striatum, but not the overlying cortex. The number of viable neurons was significantly increased after 1 and 3 days of reperfusion both in Ad-GDNF and Ad-CNTF as compared with vehicle or Ad-dE1 treated animals, whereas the number of injured cells was significantly reduced, as shown by cresyl violet staining, terminal transferase biotinylated-dUTP nick end-labeling (TUNEL), and immunocytochemistry for activated caspase-3. Interestingly, the protective effects of Ad-GDNF were similarly strong in areas of the striatum adjacent and remote of the adenoviral infusion site, while Ad CNTF showed pronounced rescue effects in the surrounding, but rather little effects distant to the infusion. The present study demonstrates that adenoviral delivery of neurotrophic factors may be a useful tool for the treatment of focal ischemia. PMID- 11493031 TI - Beta-amyloid(1-42)-induced cholinergic lesions in rat nucleus basalis bidirectionally modulate serotonergic innervation of the basal forebrain and cerebral cortex. AB - Ample experimental evidence suggests that beta-amyloid (A beta), when injected into the rat magnocellular nucleus basalis (MBN), impels excitotoxic injury of cholinergic projection neurons. Whereas learning and memory dysfunction is a hallmark of A beta-induced cholinergic deficits, anxiety, or hypoactivity under novel conditions cannot be attributed to the loss of cholinergic MBN neurons. As mood-related behavioral parameters are primarily influenced by the central serotonergic system, in the present study we investigated whether A beta(1-42) toxicity in the rat MBN leads to an altered serotonergic innervation pattern in the rat basal forebrain and cerebral cortex 7 days postsurgery. A beta infusion into the MBN elicited significant anxiety in the elevated plus maze. A beta toxicity on cholinergic MBN neurons, expressed as the loss of acetylcholinesterase-positive cortical projections, was accompanied by sprouting of serotonergic projection fibers in the MBN. In contrast, the loss of serotonin positive fiber projections, decreased concentrations of both serotonin and 5 hydroxyindoleacetic acid, and decline of cortical 5-HT(1A) receptor binding sites indicated reduced serotonergic activity in the somatosensory cortex. In conclusion, the A beta-induced primary cholinergic deficit in the MBN and subsequent cortical cholinergic denervation bidirectionally modulate serotonergic parameters in the rat basal forebrain and cerebral cortex. We assume that enhanced serotonin immunoreactivity in the damaged MBN indicates intrinsic processes facilitating neuronal recovery and cellular repair mechanisms, while diminished cortical serotonergic activity correlates with the loss of the subcortical cholinergic input, thereby maintaining the balance of neurotransmitter concentrations in the cerebral cortex. PMID- 11493032 TI - Do fits really beget fits? The effect of previous epileptic activity on the subsequent induction of the tetanus toxin model of limbic epilepsy in the rat. AB - The effect of pretreatment with either tetanus toxin (in ventral hippocampus) or kainic acid (into dorsal hippocampus, with or without suppression of seizures by phenobarbital) on the subsequent development of epilepsy in rats injected with tetanus toxin (into ventral hippocampus) has been studied. Both treatments advanced the timing of the development of the subsequent epilepsy by a few days but did not affect the severity of the syndrome. The fits stopped after 3 weeks in all the rats but recurred in 6 of 20 of those given kainic acid, with or without phenobarbital, but not in those given only tetanus toxin. It is concluded that while fits make the brain more sensitive to a further epileptogenic stimulus they do not themselves increase their severity or persistence. It is the destruction of the CA3/4 area of the hippocampus which results in this advance and in the predisposition to permanent epilepsy. PMID- 11493033 TI - Decreased MK-801 binding in discrete hippocampal regions of prion-infected mice. AB - The neurochemical alterations associated with neurodegeneration in prion diseases are not well defined. It is therefore of interest to study the influence of prion infection on messenger molecules and their receptors. In the present study we have analyzed the possible involvement of NMDA receptors in prion-infected mice using ligand binding autoradiography and iodinated MK-801, a noncompetitive NMDA antagonist. The results show a reduced binding of MK-801 in discrete regions of hippocampus at 110 days after infection, that is before the appearance of behavioral symptoms. In addition, early transient increases in MK-801 binding were observed in several layers. The exact neuroanatomical correlate of these changes in MK-801 binding, as well as its functional significance in relation to prion symptomatology, remain to be analyzed. PMID- 11493034 TI - The D355V mutation decreases EGR2 binding to an element within the Cx32 promoter. AB - Mutations in the early growth response 2 (EGR2) gene are associated with some forms of Charcot--Marie--Tooth disease (CMT) and other demyelinating neuropathies. These mutations modify the EGR2 binding to specific DNA sequences suggesting a role in the transcriptional control of myelination-specific genes. Here we show that the D355V mutation, associated with a CMT case combining axonal and demyelinating abnormalities, reduces three times the affinity of EGR2 to its consensus sequence and ten times its affinity to a sequence in the human Cx32 promoter. These findings could indicate that this EGR2 mutation leads to the development of CMT1 through the transcriptional deregulation of Cx32 gene. PMID- 11493035 TI - Distribution and modulation of histamine H(3) receptors in basal ganglia and frontal cortex of healthy controls and patients with Parkinson's disease. AB - Parkinson's disease (PD) is a brain degenerative disorder with unknown etiology, and specific degeneration of mesencephalic dopaminergic cells is a morphological manifestation of the disease. The central histaminergic system appears to be activated in PD, since the histaminergic innervation is increased in the substantia nigra. The aim of the present study was to investigate the expression and function of histamine H(3) receptors in PD, using receptor mRNA in situ hybridization with oligonucleotide probes, receptor binding assay with a specific radioactive agonist, and GTP-gamma-[(35)S]-binding assay as a tool to study the activation of the receptor G-protein. H(3) receptor binding sites were detected using N-alpha-methylhistamine autoradiography in the basal ganglia and cortex, being most abundant in the substantia nigra and striatum. In PD substantia nigra we detected an increase of the receptor binding density. In situ hybridization study of the receptor mRNA revealed prominent sites of H(3) receptor synthesis in the putamen, cortex, and globus pallidus, whereas very low mRNA expression was seen in the substantia nigra. In the PD pallidum externum, H(3) receptor mRNA expression was elevated as compared with the normal brains. GTP-gamma-[(35)S] binding assay did not reveal any significant difference between PD and normal brains, although the density values in PD substantia nigra tended to be lower than in the normal brain, and density values in PD striatum were higher. The dopaminergic neurons did not express significant amount of H(3) receptor mRNA, suggesting that the effects of H(3) receptor-mediated modulation of dopamine release are indirect. Our data indicates modulation of the histamine H(3) receptor in PD at the level of the mRNA expression in the striatum and receptor density in the substantia nigra. The receptor activity seems to be unchanged or decreased, as revealed by GTP-gamma-[(35)S]-binding assay. Modulation of the histamine H(3) receptor may influence the activity of other neurotransmitter systems, e.g., the GABAergic one, in the substantia nigra. PMID- 11493036 TI - Endoproteolysis of the ER stress transducer ATF6 in the presence of functionally inactive presenilins. AB - Presenilin (PS) proteins facilitate endoproteolysis of selected type I transmembrane proteins such as the Alzheimer's disease (AD) associated beta Amyloid precursor protein (beta APP) and Notch. beta APP is cleaved within its transmembrane domain by an aspartyl protease activity termed gamma-secretase, which may be identical with PS1 and PS2. Notch also undergoes a PS-dependent intramembraneous proteolysis. A similar gamma-secretase-like cleavage may also occur with IRE1 and ATF6, two signaling molecules of the unfolded protein response (UPR) that may require PSs for their activation. Here, we have analyzed whether ATF6 cleavage requires a PS-dependent gamma-secretase activity and whether inhibition of gamma-secretase activity would affect the UPR. Endoproteolysis of ATF6 was observed in the presence of the highly potent gamma secretase inhibitor L-685,458. ATF6 processing also occurred in the presence of functionally inactive dominant negative mutants of PS1 (PS1 D385N) and PS2 (PS2 D366A) that do not support endoproteolysis of beta APP and Notch. Our results therefore demonstrate that ATF6 is not a substrate for PS mediated gamma secretase-like endoproteolysis. This finding indicates that gamma-secretase inhibitors, which are currently developed as therapeutic agents to lower the A beta burden in brains of AD patients, do not interfere with the UPR response. PMID- 11493037 TI - What factors are associated with local enforcement of laws banning illegal tobacco sales to minors? A study of 182 law enforcement agencies in California. AB - OBJECTIVES: The aims of this study were to understand the attitudes of local law enforcement staff regarding policies to reduce youth access to tobacco, to determine what proportion of these agencies have conducted unannounced compliance checks in the past year, and to examine factors associated with conducting compliance checks. METHODS: A written questionnaire was completed by 182 law enforcement officials representing 200 cities and counties in California (some officials represented multiple jurisdictions). Logistic regression analyses were conducted to identify factors associated with conducting compliance checks. RESULTS: Only 36% of local enforcement agencies reported conducting one or more compliance checks in the previous year. Agencies were more likely to conduct compliance checks if they perceived fewer barriers to enforcement, reported more frequent collaboration with other community agencies to enforce youth access laws, and believed that youth access to tobacco is a problem in their community. CONCLUSIONS: Because active enforcement of youth access laws using unannounced compliance checks has been shown to reduce the rate of illegal tobacco sales to minors and may reduce youth smoking, efforts to increase the level of enforcement should be promoted. These study results may be used to inform efforts to encourage local agencies to enforce existing youth access laws. PMID- 11493038 TI - Page for patients. Helping make healthy legislative changes. PMID- 11493039 TI - Factors associated with daily smoking among Israeli adolescents: a prospective cohort study with a 3-year follow-up. AB - BACKGROUND: A prospective, 3-year cohort study of smoking habits was conducted among 9th grade pupils in Israel. METHODS: A self-administered questionnaire was answered by pupils in the 9th grade and again in the 11th and 12th grades. RESULTS: The baseline questionnaire was answered by 748 9th graders. In the 11th grade 448 (60%) completed the questionnaire for the second time, and in the 12th grade 388 (52%) completed the questionnaire. In all, 312 pupils (42% of the original cohort) completed all three questionnaires. There were no significant sociodemographic differences between these 312 and the original 748. Eight 9th graders (2.6%) were active smokers (at least one cigarette daily for the last month) compared with 64 (20.5%) in the 11th grade (P < 0.0001 vs 9th grade) and 70 (22.4%) in the 12th grade (P = 0.34 vs 11th grade). Most pupils knew the health hazards of smoking. Variables associated with smoking in the 11th grade included religiosity (religion was associated with lower smoking rates, P = 0.07), past experimentation with smoking (P < 0.0001), smoking among family members (P < 0.01), perceived future smoking status (P < 0.001), self-image (P < 0.001), influence of a teacher (P = 0.07) or celebrity (P < 0.05), and effect of peer pressure (P < 0.01). These results were similar for the 12th grade students except for active or previous smoking by gender (a significantly greater proportion of females than males, P = 0.04). CONCLUSIONS: The study design reduces the potential bias of follow-up attrition on assessment of predictors for smoking initiation. Gender, social modeling and peer pressure, past experimentation with smoking, smoking among family members, role models, and self image were associated with smoking. These factors should be emphasized in intervention programs. PMID- 11493040 TI - Process evaluation of a clinical preventive nutrition intervention. AB - BACKGROUND: We report process data on the feasibility of delivering a clinical preventive nutrition intervention that was effective in increasing participants' consumption of fruits and vegetables. We also examine relationships between process variables and study outcomes. METHODS: We randomly assigned six practice sites in a managed care organization to a dietary intervention or control condition. We invited adults 18 years of age or older scheduled for routine health visits within the subsequent 2 months to participate. Of the 566 patients we contacted from the intervention sites, 230 (41%) enrolled. From the control sites, we contacted 617, and 274 (44%) enrolled. Intervention participants received a tailored letter providing feedback on their consumption of target foods together with recommendations for improvement, stage-matched nutrition education booklets, a diet-health endorsement from their primary care providers (PCPs), and two motivational counseling telephone calls. Of enrollees, 195 (85%) in the intervention group and 252 (92%) in the control group returned the final survey 3 months later. RESULTS: Seventy-one percent of both participants and PCPs reported that the PCPs had discussed the relationship between diet and health at their visit. Fifty-seven percent of participants and 62% of PCPs reported that they discussed the complete diet-health endorsement, which included: (1) acknowledgment of the relationship between diet and health and (2) tailored study recommendations. The inclusion of both parts of the diet-health endorsement, but not the length of time spent, appeared to correlate with healthful outcomes. CONCLUSIONS: These process data suggest that the brief PCP diet-health endorsement contributed to the intervention effect on fruits and vegetables. PMID- 11493041 TI - EatSmart: efficacy of a multifaceted preventive nutrition intervention in clinical practice. AB - BACKGROUND: We evaluated a multifaceted preventive nutrition intervention to improve dietary habits among adult primary care patients. METHODS: We randomized six group practices in a managed care organization. Participants completed baseline and 3-month follow-up surveys. The intervention comprised (1) mailed personalized dietary recommendations and educational booklets focusing on fruits and vegetables, red meat, and dairy foods, tailored to patients' baseline intake and stage of readiness to change eating behaviors; (2) verbal endorsement by the primary care provider of the benefits of these recommendations; and (3) two motivational counseling sessions with telephone counselors to set dietary goals. RESULTS: Among the 230 subjects in the intervention group and 274 in the control group, mean age was 54 years, 70% were female, and 91% were white and 7% African American. Eighty-nine percent of the participants completed the follow-up survey. We measured change in intake of foods using results from the baseline and follow up food frequency questionnaires. Using an intention-to-treat analysis and adjusting for age, sex, race, and baseline intake, the change in fruit and vegetable intake in the intervention group was 0.6 (95% confidence interval 0.3, 0.8) servings/day higher than in the control group. There was no intervention effect on red meat and dairy products. CONCLUSIONS: Tailored clinical nutrition interventions that combine brief physician endorsement with practice supports may be effective in changing patients' eating behavior. PMID- 11493042 TI - Risk factors for sudden unexpected death among workers: a nested case-control study in central Japan. AB - BACKGROUND; Few studies have focused on sudden death among apparently healthy workers, and the risk factors have not been fully discussed. METHODS: A nested case-control study was conducted among 164,017 male employees receiving annual medical checkups in Japan. Most recent medical checkup data of 242 sudden death victims (mean age, 48.0 years) were compared with corresponding data of 505 age-, workplace-, and job-type-matched male controls. Odds ratios (ORs) and their 95% confidence intervals (CIs) for each variable were calculated by logistic regression. RESULTS: OR (95% CI) significantly increased with advancing blood pressure, reaching 6.6 (3.4-13.1) for systolic blood pressure > or =160 mm Hg relative to that <120 mm Hg. Hypo-HDL-cholesterolemia, hyperuricemia, increased aminotransferases, and abnormal urinary findings were associated with the risk in a dose-dependent manner. The presence of arrhythmias and ST-T abnormalities as well as abnormal Q waves on electrocardiograms yielded a 3.5 to 4.8 times greater risk of sudden death. As for lifestyles, heavy smoking was a positive, and light drinking was a negative risk factor. Multivariate analysis revealed that hypertension, proteinuria, glucosuria, arrhythmias, ST-T abnormalities, and light drinking were independent predictors for sudden death. CONCLUSION: These findings suggest that periodic medical checkups can help to predict and prevent employee sudden death. PMID- 11493043 TI - Hormone replacement therapy use and menopausal symptoms among women participating in a behavioral lifestyle intervention. AB - BACKGROUND: The decision to take hormone replacement therapy (HRT) is a choice many women encounter when entering menopause. The purpose of this study was to examine the choice to take HRT while participating in a lifestyle intervention to reduce cardiovascular risk through the menopause. METHODS: The Women's Healthy Lifestyle Project is a randomized clinical trial designed to examine whether a behavioral lifestyle intervention can decrease the expected rise in cardiovascular risk through the menopause. Participants (N = 535) completed questionnaires and were interviewed regarding menopausal symptoms, menopausal status, hot flashes, and HRT use at baseline and 54 months. RESULTS: The intervention was successful in preventing risk elevation through the 54-month visit. At the final visit, there was no difference between the intervention and control groups in the percentage who had become postmenopausal (32.9% vs 35.0%, respectively), there was no difference between control and intervention with HRT use, with 31.2% reporting use of HRT, and there was no difference between groups with menopausal symptoms. The women started HRT an average of 6 months after they missed a period. Baseline risk factors did not predict HRT use at the 54-month visit. CONCLUSIONS: A sizable number of women reported HRT use. The decision to use HRT was not influenced by the lifestyle intervention or their baseline cardiovascular risk, and these women started HRT very early in the peri- to postmenopause. Further, weight loss in the perimenopause did not affect menopausal symptoms. PMID- 11493044 TI - Determinants of acceptance of a community-based program for the prevention of falls and fractures among the elderly. AB - BACKGROUND: Low-energy fractures among the elderly may be prevented by measures aimed at reducing the risk of falling or increasing the strength of the skeleton. Acceptance of these interventions in the target population is necessary for their success. METHODS: The total elderly population in a Danish municipality 7,543 community-dwelling persons aged 66+ years, were offered participation in one of three intervention programs: 2,550 persons were offered a home safety inspection, evaluation of prescribed medicine, and identification of possible health and food problems (Program I); 2,445 persons were offered 1000 mg of elemental calcium and 400 IU (10 microg) of vitamin D(3) per day in combination with evaluation of prescribed medicine (Program II); and 2,548 persons were offered a combination of the two programs (Program III). Acceptance was defined as willingness to receive an introductory visit by a nurse. RESULTS: Acceptance of Program I was 50%; of Program II, 56% (P < 0.00005 as contrasted with Program I); and of Program III, 46% (P < 0.005). Acceptance was associated with gender (females, 53%; males, 47%) and did not change from ages 66 to 84 but decreased significantly after the age of 85. Widows aged 66-84 had the highest acceptance (57%) and never married males aged 66-84 the lowest (30%). An important determinant, however, was the individual social service center that communicated the specific program. Acceptance varied from 39 to 66% between the social centers. CONCLUSIONS: Acceptance of a fall and fracture prevention program varies with intervention type; with gender, age, and social status of the target population; and with the motivation and attitude of the health workers involved in the implementation of the program. PMID- 11493045 TI - Walking and resting blood pressure in adults: a meta-analysis. AB - BACKGROUND: The purpose of this study was to examine the effects of walking on resting systolic and diastolic blood pressure in adults. METHODS: A total of 24 primary outcomes from 16 studies and 650 subjects (410 exercise, 240 control) met the criteria for inclusion: (1) randomized and nonrandomized controlled trials, (2) walking as the only intervention, (3) subjects apparently sedentary, (4) adult humans > or =18 years of age, (5) English-language studies published between January 1966 and December 1998, (6) resting blood pressure assessed, (7) training studies > or =4 weeks. RESULTS: Using a random effects model, statistically significant decreases of approximately 2% were found for both resting systolic and diastolic blood pressure (systolic, mean +/- SEM = -3 +/- 1 mm Hg, 95% confidence interval: -5 to -2 mm Hg; diastolic, mean +/- SEM = -2 +/- 1 mm Hg, 95% confidence interval: -3 to -1 mm Hg). CONCLUSION: Walking exercise programs reduce resting blood pressure in adults. PMID- 11493046 TI - Health beliefs and endoscopic screening for colorectal cancer: potential for cancer prevention. AB - BACKGROUND: Colorectal cancer can be prevented through endoscopic removal of adenomatous polyps. Because screening endoscopy rates are low, it is critical to identify correlates of screening behavior that are amenable to interventions to improve screening rates. Our purpose was to identify the correlates of endoscopic screening among persons at risk for colorectal cancer. METHODS: We surveyed 1,160 healthy, adult, first-degree relatives of colorectal cancer patients in 583 kindreds, for a 43% response rate. Using multiple logistic regression analysis, we tested the association between screening behavior and perceived risk for colorectal cancer, the belief that colorectal cancer can be prevented, demographic factors, strength of family history, and practical barriers to screening. RESULTS: Persons screened at least once were older, were male, had stronger family histories, had a regular doctor, and had health insurance. After these fixed factors were accounted for, the belief that colorectal cancer can be prevented and higher perceived risk were associated with significantly greater odds of screening. CONCLUSIONS: This study establishes the need for trials evaluating the cancer prevention potential of the link between screening behavior and health beliefs. Physicians must be aware of their patients' family colorectal cancer history and recommend appropriate endoscopic screening for those at increased risk, particularly women. Patients should be educated about their cancer risk and about preventing colorectal cancer. PMID- 11493047 TI - Lack of public awareness toward alcohol consumption as risk factor for oral and pharyngeal cancers. PMID- 11493048 TI - Influence of the dynamic positions of cations on the structure of the DNA minor groove: sequence-dependent effects. AB - Different models for minor groove structures predict that the conformation is essentially fixed by sequence and has an influence on local ion distribution or alternatively that temporal positions of ions around the minor groove can affect the structure if they neutralize cross-strand phosphate charges. Our previous studies show that the minor groove in an AATT dodecamer responds to local sodium ion positions and is narrow when ions neutralize cross-strand phosphate-phosphate charges [J. Am. Chem. Soc. 2000, 122, 10513-10520]. Previous results from a number of laboratories have shown that G-tracts often have a wider minor groove than A-tracts, but they do not indicate whether this is due to reduced flexibility or differences in ion interactions. We have undertaken a molecular dynamics study of a d(TATAGGCCTATA) duplex to answer this question. The results show that the G-tract has the same amplitude of minor groove fluctuations as the A-tract sequence but that it has fewer ion interactions that neutralize cross strand phosphate charges. These results demonstrate that differences in time average groove width between A- and G-tracts are due to differences in ion interactions at the minor groove. When ions neutralize the cross-strand phosphates, the minor groove is narrow. When there are no neutralizing ion interactions, the minor groove is wide. The population of structures with no ion interactions is larger with the GGCC than with the AATT duplex, and GGCC has a wider time-average minor groove in agreement with experiment. PMID- 11493049 TI - Change in spin state and enhancement of redox reactivity of photoexcited states of aromatic carbonyl compounds by complexation with metal ion salts acting as Lewis acids. Lewis acid-catalyzed photoaddition of benzyltrimethylsilane and tetramethyltin via photoinduced electron transfer. AB - The lowest excited state of aromatic carbonyl compounds (naphthaldehydes, acetonaphthones, and 10-methylacridone) is changed from the n,pi triplet to the pi,pi singlet which becomes lower in energy than the n,pi triplet by the complexation with metal ions such as Mg(ClO(4))(2) and Sc(OTf)(3) (OTf = triflate), which act as Lewis acids. Remarkable positive shifts of the one electron reduction potentials of the singlet excited states of the Lewis acid carbonyl complexes (e.g., 1.3 V for the 1-naphthaldehyde-Sc(OTf)(3) complex) as compared to those of the triplet excited states of uncomplexed carbonyl compounds result in a significant increase in the redox reactivity of the Lewis acid complexes vs uncomplexed carbonyl compounds in the photoinduced electron-transfer reactions. Such enhancement of the redox reactivity of the Lewis acid complexes leads to the efficient C-C bond formation between benzyltrimethylsilane and aromatic carbonyl compounds via the Lewis-acid-promoted photoinduced electron transfer. The quantum yield determinations, the fluorescence quenching, and direct detection of the reaction intermediates by means of laser flash photolysis experiments indicate that the Lewis acid-catalyzed photoaddition reactions proceed via photoinduced electron transfer from benzyltrimethylsilane to the singlet excited states of Lewis acid-carbonyl complexes. PMID- 11493050 TI - Catalytic asymmetric ring-opening metathesis/cross metathesis (AROM/CM) reactions. Mechanism and application to enantioselective synthesis of functionalized cyclopentanes. AB - Studies regarding the first examples of catalytic asymmetric ring-opening metathesis (AROM) reactions are detailed. This enantioselective cleavage of norbornyl alkenes is followed by an intermolecular cross metathesis with a terminal olefin partner; judicious selection of olefin is required so that oligomerization and dimerization side products are avoided. Results outlined herein suggest that the presence of suitably positioned heteroatom substituents may be critical to reaction efficiency. Mo-catalyzed tandem AROM/CM affords functionalized cyclopentyl dienes in >98% ee and >98% trans olefin selectivity; both secondary and tertiary ether products can be obtained. The examples provided include the catalytic synthesis of an optically pure cyclopentyl epoxide and dimethyl acetal. Mechanistic studies suggest that it is the more substituted benzylidene or silylated alkylidenes that are involved in the catalytic process (vs the corresponding Mo-methylidenes). Although electron rich benzylidenes react more efficiently, the derived electron poor Mo complexes promote AROM/CM transformations as well; alkylidenes that bear a boron substituent are unreactive. PMID- 11493051 TI - Palladium catalysis for the synthesis of hydrophobic C-6 and C-2 aryl 2' deoxynucleosides. Comparison of C-C versus C-N bond formation as well as C-6 versus C-2 reactivity. AB - Suzuki-Miyaura cross-coupling of haloaromatic compounds with arylboronic acids provides a simple entry to biaryl systems. Despite its ease, to date, there are no detailed investigations of this procedure for deoxynucleoside modification. As shown in this study, a wide variety of C-6 arylpurine 2'-deoxyriboside (C-6 aryl 2'-deoxynebularine analogues) and C-2 aryl 2'-deoxyinosine analogues can be conveniently prepared via the Pd-mediated cross-coupling of arylboronic acids with the C-6 halonucleosides, 6-bromo- or 6-chloro-9[2-deoxy-3,5-bis-O-(tert butyldimethylsilyl)-beta-D-erythro-pentofuranosyl]purine (1 and 2), and the C-2 halonucleoside, 2-bromo-O(6)-benzyl-3',5'-bis-O-(tert-butyldimethylsilyl)-2' deoxyinosine (3). Although bromonucleoside 1 proved to be a good substrate for the Pd-catalyzed Suzuki-Miyaura cross-couplings, we have noted that for several C 6 arylations, the chloronucleoside 2 provides superior coupling yields. Also described in this study is a detailed evaluation of catalytic systems that led to optimal product recoveries. Finally, a comparison of the C-C and C-N bond-forming reactions of deoxynucleosides is also reported. On the basis of this comparison, we provide evidence that C-N bond formation at the C-6 position, leading to N aryl 2'-deoxyadenosine analogues, is more sensitive to the ligand used, whereas C C bond-forming reactions at the same position are not. In contrast to the ligand dependency exhibited in C-N bond formation at the C-6 position, comparable reactions at the C-2 position of purine deoxynucleosides proceed with less sensitivity to the ligand used. PMID- 11493052 TI - High-pressure bulk synthesis of crystalline C(6)N(9)H(3).HCl: a novel c(3)n(4) graphitic derivative. AB - A novel carbon nitride compound, structurally related to the proposed graphitic phase of C(3)N(4), has been synthesized in a bulk well-crystallized form. The new material, with stoichiometry C(6)N(9)H(4)Cl, was prepared through a solid-state reaction of 2,4,6-triamino-1,3,5-triazine with 2,4,6-trichloro-1,3,5-triazine at 1.0-1.5 GPa and 500-550 degrees C and also through a self-reaction of 2-amino-4,6 dichloro-1,3,5-triazine at similar conditions. X-ray and electron diffraction measurements on the yellowish compound indicate a hexagonal space group (P6(3)/m) with cell parameters of a = 8.4379(10) A and c = 6.4296(2) A. This new compound possesses a two-dimensional C(6)N(9)H(3) framework that is structurally related to the hypothetical P6m2 graphitic phase of C(3)N(4), but with an ordered arrangement of C(3)N(3) voids. The large voids in the graphene sheets are occupied by chloride ions with an equivalent number of nitrogen atoms on the framework protonated for charge balance. The composition of the sample was determined by bulk chemical analysis and confirmed by electron energy loss (EELS) spectroscopy. The chemical and structural model is consistent with bulk density measurements and with the infrared and (13)C NMR spectra. This work represents the first bulk synthesis of a well-characterized and highly crystalline material containing a continuous network of alternating carbon and nitrogen atoms. PMID- 11493053 TI - Synthesis of organic monolayer-stabilized copper nanocrystals in supercritical water. AB - When water is heated and pressurized above the critical point, it becomes a suitable solvent to employ organic capping ligands to control and stabilize the synthesis of nanocrystals. Without alkanethiol ligands, Cu(NO(3))(2) hydrolyzes to form polydisperse copper(II) oxide particles with diameters from 10 to 35 nm. However, in the presence of 1-hexanethiol, X-ray photoelectron spectroscopy, selected area electron diffraction, and transmission electron microscopy reveal the formation of copper nanocrystals approximately 7 nm in diameter. The use of a different precursor, Cu(CH(3)COO)(2), leads to particles with significantly different morphologies. A mechanism is proposed for sterically stabilized nanocrystal growth in supercritical water that describes competing pathways of hydrolysis to large oxidized copper particles versus ligand exchange and arrested growth by thiols to produce small monodisperse Cu nanoparticles. PMID- 11493055 TI - Exciton-mediated hydrosilylation on photoluminescent nanocrystalline silicon. AB - A novel white light-promoted reaction using photoluminescent nanocrystalline silicon enables the hydrosilylation of alkenes and alkynes, providing stabilization of the porous silicon without significant loss of the photoemissive qualities of the material. Photopatterning and lithographic fabrication of isolated porous silicon structures are made possible. Experiments and observations are presented which indicate that the light promoted hydrosilylation reaction is unique to photoluminescent silicon, and does not function on nonemissive material. Hydrosilylation using a reactive center generated from a surface-localized exciton is proposed based upon experimental evidence, explaining the photoluminescence requirement. Indirect excitons formed by light absorption mediate the formation of localized electrophilic surface states which are attacked by incoming alkene or alkyne nucleophiles. Supra-band gap charge carriers have sufficient energy to react with nucleophilic alkenes and alkynes, thereupon causing Si-C bond formation, an irreversible event. The light-promoted hydrosilylation reaction is quenched by reagents that quench the light emission from porous silicon, via both charge transfer and energy transfer pathways. PMID- 11493054 TI - Absence of Mn-centered oxidation in the S(2) --> S(3) transition: implications for the mechanism of photosynthetic water oxidation. AB - A key question for the understanding of photosynthetic water oxidation is whether the four oxidizing equivalents necessary to oxidize water to dioxygen are accumulated on the four Mn ions of the oxygen-evolving complex (OEC), or whether some ligand-centered oxidations take place before the formation and release of dioxygen during the S(3) --> [S(4)] --> S(0) transition. Progress in instrumentation and flash sample preparation allowed us to apply Mn Kbeta X-ray emission spectroscopy (Kbeta XES) to this problem for the first time. The Kbeta XES results, in combination with Mn X-ray absorption near-edge structure (XANES) and electron paramagnetic resonance (EPR) data obtained from the same set of samples, show that the S(2) --> S(3) transition, in contrast to the S(0) --> S(1) and S(1) --> S(2) transitions, does not involve a Mn-centered oxidation. On the basis of new structural data from the S(3)-state, manganese mu-oxo bridge radical formation is proposed for the S(2) --> S(3) transition, and three possible mechanisms for the O-O bond formation are presented. PMID- 11493056 TI - Fluorescent sensors for Zn(2+) based on a fluorescein platform: synthesis, properties and intracellular distribution. AB - Two new fluorescent sensors for Zn(2+) that utilize fluorescein as a reporting group, Zinpyr-1 and Zinpyr-2, have been synthesized and characterized. Zinpyr-1 is prepared in one step via a Mannich reaction, and Zinpyr-2 is obtained in a multistep synthesis that utilizes 4',5'-fluorescein dicarboxaldehyde as a key intermediate. Both Zinpyr sensors have excitation and emission wavelengths in the visible range ( approximately 500 nm), dissociation constants (K(d1)) for Zn(2+) of <1 nM, quantum yields approaching unity (Phi = approximately 0.9), and cell permeability, making them well-suited for intracellular applications. A 3- to 5 fold fluorescent enhancement is observed under simulated physiological conditions corresponding to the binding of the Zn(2+) cation to the sensor, which inhibits a photoinduced electron transfer (PET) quenching pathway. The X-ray crystal structure of a 2:1 Zn(2+):Zinpyr-1 complex has also been solved, and is the first structurally characterized example of a complex of fluorescein substituted with metal binding ligands. PMID- 11493057 TI - Correlation between molecular structure and helicity of induced chiral nematics in terms of short-range and electrostatic-induction interactions. The case of chiral biphenyls. AB - The helical structure of the chiral nematic phases induced by chiral dopants in nematic solvents provides a macroscopic image of the molecular chirality of the dopant promoted by the orientational order. Chiral biphenyls are challenging systems because their twisting ability shows a strong dependence on the molecular structure, which does not conform to empirical correlation rules. This points out the need for adequate interpretative tools, able to establish a link between molecular properties and macroscopic response. In this paper the twisting ability of chiral biphenyls is reviewed, by reporting examples taken from the literature together with some new experimental results. The microscopic origin of the observed behavior is explained in terms of chirality and anisotropy of short range and electrostatic-induction interactions. These are described, respectively, by a shape model and a reaction field method, having the common characteristics of a realistic representation of the structure and properties of the chiral dopants in terms of molecular surface, atom charges, and distributed polarizabilities. PMID- 11493058 TI - Dynamics of an inclusion complex of chloroform and cryptophane-E: evidence for a strongly anisotropic van der Waals bond. AB - The motional dynamics of a van der Waals inclusion complex of cryptophane-E and chloroform has been investigated by a combined NMR exchange and relaxation study. The kinetics of exchange of chloroform between the bulk solution and the complex was investigated by means of proton EXSY measurements. The carbon-13 relaxation of the cryptophane-E host and of the bound chloroform guest was analyzed using the Lipari-Szabo "model-free" approach. For interpretation of the carbon-13 relaxation measurements for chloroform, the chemical-exchange process of complex formation and dissociation had to be taken into account in terms of the modified Bloch equations. It was found that the complex behaves as a single molecule without any significant guest chloroform motion inside the host's cavity. PMID- 11493059 TI - Conformational switching and exciton interactions in hemicyanine-based bichromophores. AB - Conformational changes in two hemicyanine-based bichromophores were demonstrated by varying the polarity as well as temperature of the medium. Dramatic changes in the ground and excited singlet state properties were observed upon folding of the bichromophores, due to the formation of intramolecular aggregates of H-type. These aspects were studied, in detail, using steady-state absorption and time resolved fluorescence spectroscopy. Time-resolved fluorescence studies indicate that both the bichromophores exhibit a monoexponential decay, with a short lifetime, in mixed toluene-CH(2)Cl(2) solvents having lower proportions of toluene. Interestingly, biexponential decay with short and long-lived species was observed at higher proportions of toluene, due to the presence of unfolded and folded forms. Folding results in the intramolecular stacking of the chromophores which restrict their torsional dynamics, leading to a longer lifetime. Upon laser excitation, the folded form of the bichromophore undergoes rapid conformational changes, due to photoinduced thermal dissociation. PMID- 11493060 TI - Geometry, reduction potential, and reorganization energy of the binuclear Cu(A) site, studied by density functional theory. AB - The dimeric Cu(A) site found in cytochrome c oxidase and nitrous oxide reductase has been studied with the density functional B3LYP method. We have optimized the structure of the realistic (Im)(S(CH(3))(2))Cu(SCH(3))(2)Cu(Im)(CH(3)CONHCH(3)) model in the fully reduced, mixed-valence, and fully oxidized states. The optimized structures are very similar to crystal structures of the protein, which shows that the protein does not strain the site significantly. Instead, inorganic model complexes of the protein site are strained by the macrocyclic connections between the ligand models. For the mixed-valence (Cu(I)+Cu(II)) state, two distinct equilibrium structures were found, one with a short Cu-Cu distance, 248 pm, similar to the protein structure, and one with a longer distance, 310 pm, similar to what is found in inorganic models. In the first state, the unpaired electron is delocalized over both copper ions, whereas in the latter, it is more localized to one of the ions. The two states are nearly degenerate. The potential energy surfaces for the Cu-Cu, Cu-S(Met), and Cu-O interactions are extremely flat. In fact, all three distances can be varied between 230 and 310 pm at an expense in energy of less than 8 kJ/mol, which explains the large variation observed in crystal structures for these interactions. Inclusion of solvation effects does not change this significantly. Therefore, we can conclude that a variation in these distances can change the reduction potential of the Cu(A) site by at most 100 mV. The model complex has a reorganization energy of 43 kJ/mol, 20 kJ/mol lower than for a monomeric blue-copper site. This lowering is caused by the delocalization of the unpaired electron in the mixed-valence state. PMID- 11493061 TI - Conformational gating of long distance electron transfer through wire-like bridges in donor-bridge-acceptor molecules. AB - A series of five donor-bridge-acceptor (DBA) molecules in which the donor is tetracene, the acceptor is pyromellitimide, and the bridge molecules are oligo-p phenylenevinylenes (OPV) of increasing length has been shown to undergo electron transfer (ET) by means of two mechanisms. When the bridge is short, strongly distance dependent superexchange dynamics dominates, whereas when the bridge is longer, bridge-assisted hopping dynamics prevails. The latter mechanism results in relatively soft distance dependence for ET in which the OPV oligomers act effectively as molecular wires. We now report studies on the critical influence that bridge dynamics have on electron transfer through these oligomers. The temperature dependence of the charge separation (CS) rates in all five molecules does not appear to obey the predictions of standard ET theories based upon the Condon approximation. All five molecules show behavior consistent with CS being "gated" by torsional motion between the tetracene donor and the first bridge phenyl ring. This is based on the near equivalence of the CS activation energies measured for all five molecules with the frequency of a known vibrational mode in 5-phenyltetracene. In the molecule containing a trans-stilbene bridge, a competition occurs between the tetracene-phenyl torsional motion and one that occurs between the vinyl group and the phenyls linked to it. This results in complex temperature-dependent CS that exhibits both activated and negatively activated regimes. The charge recombination (CR) reactions within the molecules which have the two shortest bridges, namely phenyl and trans-stilbene, show a weaker dependence on these molecular motions. The three molecules with the longest bridges all display complex temperature dependencies in both their rates of CS and CR, most likely because of the complex torsional motions, which arise from the multiple phenyl-vinyl linkages. The data show that long-distance electron transfer and therefore wire-like behavior within conjugated bridge molecules depend critically on these low-frequency torsional motions. Molecular device designs that utilize such bridges will need to address these issues. PMID- 11493062 TI - "Dip-Pen" nanolithography on semiconductor surfaces. AB - Dip-Pen Nanolithography (DPN) uses an AFM tip to deposit organic molecules through a meniscus onto an underlying substrate under ambient conditions. Thus far, the methodology has been developed exclusively for gold using alkyl or aryl thiols as inks. This study describes the first application of DPN to write organic patterns with sub-100 nm dimensions directly onto two different semiconductor surfaces: silicon and gallium arsenide. Using hexamethyldisilazane (HMDS) as the ink in the DPN procedure, we were able to utilize lateral force microscopy (LFM) images to differentiate between oxidized semiconductor surfaces and patterned areas with deposited monolayers of HMDS. The choice of the silazane ink is a critical component of the process since adsorbates such as trichlorosilanes are incompatible with the water meniscus and polymerize during ink deposition. This work provides insight into additional factors, such as temperature and adsorbate reactivity, that control the rate of the DPN process and paves the way for researchers to interface organic and biological structures generated via DPN with electronically important semiconductor substrates. PMID- 11493063 TI - High-frequency and field EPR investigation of (8,12-diethyl-2,3,7,13,17,18 hexamethylcorrolato)manganese(III). AB - High-field and frequency electron paramagnetic resonance (HFEPR) of solid (8,12 diethyl-2,3,7,13,17,18-hexamethylcorrolato)manganese(III), 1, shows that in the solid state it is well described as an S = 2 (high-spin) Mn(III) complex of a trianionic ligand, [Mn(III)C(3)(-)], just as Mn(III) porphyrins are described as [Mn(III)P(2)(-)](+). Comparison among the structural data and spin Hamiltonian parameters reported for 1, Mn(III) porphyrins, and a different Mn(III) corrole, [(tpfc)Mn(OPPh(3))], previously studied by HFEPR (Bendix, J.; Gray, H. B.; Golubkov, G.; Gross, Z. J. Chem. Soc., Chem. Commun. 2000, 1957-1958), shows that despite the molecular asymmetry of the corrole macrocycle, the electronic structure of the Mn(III) ion is roughly axial. However, in corroles, the S = 1 (intermediate-spin) state is much lower in energy than in porphyrins, regardless of axial ligand. HFEPR of 1 measured at 4.2 K in pyridine solution shows that the S = 2 [Mn(III)C(3)(-)] system is maintained, with slight changes in electronic parameters that are likely the consequence of axial pyridine ligand coordination. The present result is the first example of the detection by HFEPR of a Mn(III) complex in solution. Over a period of hours in pyridine solution at ambient temperature, however, the S = 2 Mn(III) spectrum gradually disappears leaving a signal with g = 2 and (55)Mn hyperfine splitting. Analysis of this signal, also observable by conventional EPR, leads to its assignment to a manganese species that could arise from decomposition of the original complex. The low-temperature S = 2 [Mn(III)C(3)(-)] state is in contrast to that at room temperature, which is described as a S = 1 system deriving from antiferromagnetic coupling between an S = (3/2) Mn(II) ion and a corrole-centered radical cation: [Mn(II)C(*)(2-)] (Licoccia, S.; Morgante, E.; Paolesse, R.; Polizio, F.; Senge, M. O.; Tondello, E.; Boschi, T. Inorg. Chem. 1997, 36, 1564-1570). This temperature-dependent valence state isomerization has been observed for other metallotetrapyrroles. PMID- 11493064 TI - A solid-state NMR, X-ray diffraction, and ab initio computational study of hydrogen-bond structure and dynamics of pyrazole-4-carboxylic acid chains. AB - Using high-resolution solid-state (15)N CMAS NMR, X-ray crystallography, and ab initio calculations, we have studied the structure of solid pyrazole-4-carboxylic acid (1). The crystal structure was determined at 295 and 150 K. Molecules of 1 are located on a two-fold axis, implying proton disorder of the NH and OH groups; no phase transition was observed between these two temperatures. The compound forms quasi-linear ribbons in which the molecules are linked by cyclic hydrogen bonds between pyrazole and carboxylic acid groups with disordered hydrogen-bonded protons. Crystallography is unable to decide whether the disorder is dynamic or static. NMR shows that this disorder is dynamic, that is, consisting of very fast degenerate double proton transfers between two rapidly interconverting O-H.N and O.H-N hydrogen bridges. However, at low temperature, NMR shows a proton disorder order transition where the protons are preferentially localized on given nitrogen and oxygen atoms. An amorphous phase exhibiting proton order is observed when the compound is precipitated rapidly. In this case, the defects are annealed by moderate heating. Ab initio calculations performed on oligomers of 1 show that the O-H.N hydrogen bridge is about 0.064 A shorter and less bent ( approximately 171 degrees ) than the O.H-N hydrogen bridge ( approximately 150 degrees ). For an isolated ribbon, this result leads to structures with localized protons, either to a cycle with about 200 molecules, or to a quasi-linear ribbon involving an undulated structure, or to a combination of both motifs. Only the undulated structure is compatible with the linear ribbon observed by X-ray crystallography, where the fast proton transfer in the high-temperature phase is assisted by the motions of the undulated chain. A disordered structure is assigned to the amorphous phase, which exhibits the combination of the curved and the undulated motifs. PMID- 11493065 TI - Helix formation in unsolvated peptides: side chain entropy is not the determining factor. PMID- 11493066 TI - Study of C-4 deoxygenation in the biosynthesis of desosamine: evidence implicating a novel mechanism. PMID- 11493068 TI - Facile synthesis of well-defined, biocompatible phosphorylcholine-based methacrylate copolymers via atom transfer radical polymerization at 20 degrees C. PMID- 11493067 TI - A new type of potent inhibitors of HIV-1 TAR RNA-Tat peptide binding by zinc(II) macrocyclic tetraamine complexes. PMID- 11493069 TI - CPI: a recipe for improving applicable properties of discotic liquid crystals. PMID- 11493070 TI - Conducting polymers incorporating tungsten-capped calixarenes. PMID- 11493071 TI - Isotope-edited Raman spectroscopy of proteins: a general strategy to probe individual peptide bonds with application to insulin. PMID- 11493072 TI - Approach to a single-component ferrimagnetism by organic radical crystals. PMID- 11493073 TI - Synthesis and characterization of a distonic nitrene ion: gas-phase reactivity of singlet and triplet N-phenyl-3-nitrenopyridinium ion. PMID- 11493074 TI - Metalative Reppe reaction. Organized assembly of acetylene molecules on titanium template leading to a new style of acetylene cyclotrimerization. PMID- 11493075 TI - Facile dinitrogen reduction via organometallic Tm(II) chemistry. PMID- 11493076 TI - Enantiopure dihydro-[5]-helicenequinones via Diels-Alder reactions of vinyl dihydrophenanthrenes and (SS)-2-(p-tolylsulfinyl)-1,4-benzoquinone. PMID- 11493077 TI - Photorelease of HCl from o-methylphenacyl chloride proceeds through the Z xylylenol. PMID- 11493078 TI - Frontal polymerization synthesis of temperature-sensitive hydrogels. PMID- 11493080 TI - Remarkably facile, unidirectional isomerization of titanated vinylallenes to cyclobutenyltitanium compounds. a practical construction of a cyclobutene framework. PMID- 11493079 TI - A gallium-substituted cubane-type cluster in Pyrococcus furiosus ferredoxin. PMID- 11493081 TI - The structural role of cholesterol in biological membranes. PMID- 11493082 TI - "Cation flow" method: a new approach to conventional and combinatorial organic syntheses using electrochemical microflow systems. PMID- 11493083 TI - Reversal of polymerization stereoregulation in anionic polymerization of MMA by chiral metallocene and non-metallocene initiators: a new reaction pathway for metallocene-initiated MMA polymerization. PMID- 11493084 TI - Intramolecular, nucleophile-catalyzed aldol-lactonization (NCAL) reactions: catalytic, asymmetric synthesis of bicyclic beta-lactones. PMID- 11493085 TI - Phosphine ligand exchange at a phosphine lewis acceptor: the first structural characterization of homoleptic phosphinophosphonium salts. PMID- 11493086 TI - Glycine enolates: the large effect of iminium ion formation on alpha-amino carbon acidity. PMID- 11493087 TI - On the validity of the maximum hardness and minimum polarizability principles for nontotally symmetric vibrations. PMID- 11493088 TI - Enantiospecific desorption of R- and S-propylene oxide from a chiral Cu(643) surface. PMID- 11493089 TI - Self-organization of large gold nanoparticle arrays. PMID- 11493090 TI - Self-organized hybrid silica with long-range ordered lamellar structure. PMID- 11493091 TI - Doping molecular semiconductors: n-type doping of a liquid crystal perylene diimide. PMID- 11493092 TI - DNA-modified core-shell Ag/Au nanoparticles. PMID- 11493093 TI - Topical imiquimod for acyclovir-unresponsive herpes simplex virus 2 infection. PMID- 11493094 TI - Permeability barrier function of skin exposed to ionizing radiation. AB - OBJECTIVE: To characterize the epidermal permeability barrier function of skin during exposure to ionizing radiation. DESIGN: A prospective cohort study. SETTING: University hospital medical center. PATIENTS: Fifteen women receiving local radiation therapy (5000-6000 rad [50-60 Gy]) following breast-conserving surgery for breast cancer. MAIN OUTCOME MEASURES: Clinical symptoms and transepidermal water loss (TEWL). RESULTS: Epidermal permeability barrier function is impaired in patients who exhibit clinical signs of radiation dermatitis. The functional damage to the stratum corneum induced by ionizing radiation occurs with a delayed course, starting within a mean period of 11 days and reaching maximal values after a mean period of 27 days (range, 13-75 days). The onset of TEWL increase precedes the onset of radiation dermatitis and the maximal TEWL measurements precede the peak of skin changes. Patients with an early onset of TEWL increase show a longer duration of skin symptoms. CONCLUSIONS: Skin changes caused by radiation dermatitis are associated with an increase in TEWL. The barrier impairment is comparable to the changes observed with UV radiation exposure but exhibits an even more delayed course. Our results suggest that preservation of the epidermal permeability barrier function by topical treatment may ameliorate radiation dermatitis. PMID- 11493095 TI - Nail lichen planus in children: clinical features, response to treatment, and long-term follow-up. AB - OBJECTIVE: To report clinical features, response to treatment, and long-term follow-up of nail lichen planus in children. DESIGN: Retrospective study involving 15 children with nail lichen planus. SETTING: Outpatient consultation for nail disorders at the Department of Dermatology of the University of Bologna, Bologna, Italy. PATIENTS OR OTHER PARTICIPANTS: We diagnosed nail lichen planus in 15 children younger than 12 years, including 10 children with typical nail matrix lesions, 2 children with 20-nail dystrophy (trachyonychia), and 3 children with idiopathic atrophy of the nails. Only 2 of the 15 children had oral lichen planus; none had cutaneous lesions. A nail biopsy confirmed the diagnosis in all cases. INTERVENTION: Intramuscular triamcinolone acetonide, 0.5 to 1 mg/kg per month, was prescribed to children with typical nail lichen planus and prolonged from 3 to 6 months until the proximal half of the nail was normal. No treatment was prescribed to patients with 20-nail dystrophy or idiopathic atrophy of the nails. RESULTS: Treatment with systemic corticosteroids was effective in curing typical nail lichen planus. Two children experienced a recurrence of the disease during the follow-up. Recurrences were always responsive to therapy. The 2 children with 20-nail dystrophy improved without any therapy. Nail lesions caused by idiopathic atrophy of the nails remained unchanged during the follow-up period. CONCLUSIONS: Nail lichen planus in children is not rare but probably underestimated. It often presents with atypical clinical features such as 20-nail dystrophy or idiopathic atrophy of the nails. PMID- 11493096 TI - Saline spa water or combined water and UV-B for psoriasis vs conventional UV-B: lessons from the Salies de Bearn randomized study. AB - OBJECTIVE: To study the effects of UV-B therapy and saline spa water given alone or in combination for the treatment of psoriasis. DESIGN: Randomized, controlled, comparative study with blinded observers. SETTING: Salies de Bearn, saline spa water center located in the southwest of France. PARTICIPANTS: Seventy-one adult patients with psoriasis with a Psoriasis Area and Severity Index (PASI) score greater than 10. INTERVENTION: Patients were randomly assigned to 1 of 3 treatments: spa water alone (group A); UV-B 311-nm phototherapy alone (group B); and a combination of the 2 therapies (group C). The 3 groups were treated on a daily basis 5 days a week for a total of 21 days. MAIN OUTCOME MEASURES: Change in PASI score from baseline as determined by an investigator blinded to randomization; variation in quality of life, adverse effects, and long-term effects (1 year after treatment). RESULTS: Four patients dropped out because of secondary effects. Efficacy was similar in groups B and C, with changes in PASI of -64% and -55%, respectively at 3 weeks. For group A, change in PASI was -29%, thus showing a minor therapeutic effect of saline spa water alone and poor efficacy compared with groups B and C (P<.001). More adverse effects were reported in groups A and C but did not reach significance. Combined saline spa water and UV-B therapy had no sparing effect on UV-B dosages. One year after treatment, no long-term benefit could be attributed specifically to a given regimen, but the patients had overall significantly better PASI scores than at baseline. CONCLUSIONS: Saline spa water alone had a minor therapeutic effect in psoriasis, and the beneficial effect of bathing to enhance phototherapy was not demonstrated. PMID- 11493097 TI - Cutaneous photodamage in Koreans: influence of sex, sun exposure, smoking, and skin color. AB - BACKGROUND: Severe wrinkles and pigmentary changes of the exposed skin indicate substantial damage due to UV radiation. Many investigators believe that the principal manifestation of photodamage in Asians is pigmentary change rather than wrinkles. However, to our knowledge, no well-designed study has investigated the characteristics of cutaneous photodamage in Asian skin. OBJECTIVE: To access the severity of wrinkles and dyspigmentation in Koreans exposed to sun and who smoked. METHODS: We developed new photographic scales for grading wrinkles and dyspigmentation in 407 Koreans to assess the severity of the wrinkles and dyspigmentation. We interviewed subjects to determine cumulative sun exposure and smoking history, and measured the skin color of individual subjects. RESULTS: Our photographic scales provided a reliable evaluation of photodamage severity in Koreans. The pattern of wrinkling in both sexes is similar, but women tended to have more severe wrinkles (prevalence odds ratio, 3.7). However, the pattern of dyspigmentation differed between the sexes. Seborrheic keratosis is the major pigmentary lesion in men, whereas hyperpigmented macules are the prominent features in women. Cigarette smoking is an independent risk factor for wrinkles, but not for dyspigmentation, in Koreans, and causes additive detrimental effects to wrinkles induced by aging and sun exposure. The constitutive skin color did not show any correlation with wrinkles or dyspigmentation. However, facultative pigmentation (sun exposure index) may reflect lifetime sun exposure, and it shows a good correlation with wrinkles in Koreans. CONCLUSION: Wrinkling is a major feature of photoaging in Koreans, as are pigmentary changes; smoking, sun exposure, and female sex are independent risk factors for wrinkles. PMID- 11493098 TI - Reliability of counting actinic keratoses before and after brief consensus discussion: the VA topical tretinoin chemoprevention (VATTC) trial. AB - OBJECTIVE: To assess the reliability of counts of actinic keratoses (AKs) and the effect of a brief joint discussion of discrepancies on that reliability. DESIGN AND INTERVENTION: Seven dermatologists independently counted AKs on the face and ears before and after a brief joint discussion of discrepancies. SETTING AND PATIENTS: A volunteer sample of 9 patients from the ongoing VA (Department of Veterans Affairs) Topical Tretinoin Chemoprevention (VATTC) Trial. All participating individuals are veterans and have had 2 or more keratinocyte carcinomas (basal or squamous cell carcinoma) in the 5 years before enrollment in the study. MAIN OUTCOME MEASURE: Standard deviation of estimates of the Poisson regression parameter for the dermatologists. RESULTS: Substantial variation was found among the dermatologists in their AK counts. The SD of the parameter estimates for the dermatologists decreased from 0.45 to 0.24 after the brief joint discussion, a 47% decrease (P =.076). The variation attributable to the dermatologists also decreased substantially (chi(2)(6) decrease, 94 to 12). CONCLUSIONS: Actinic keratoses are common, and there is a continuous spectrum of lesions that ranges from sun-damaged skin to squamous cell carcinoma in situ. Clinical distinguishing features may be difficult to delineate precisely. Counts of AK are commonly performed, but appear to be unreliable, even when performed by experienced dermatologists. Joint discussion of discrepancies may enhance the reliability of these counts, although substantial variation remains. Research that relied on these counts must be reevaluated in light of the marked variation among expert observers. Future studies should consider measures to assess and enhance reliability. PMID- 11493099 TI - Predictive model for immunotherapy of alopecia areata with diphencyprone. AB - BACKGROUND: Immunotherapy with diphencyprone (diphenylcyclopropenone) is used in the treatment of alopecia areata (AA). Response rates have varied in the literature. OBJECTIVES: To determine the efficacy of diphencyprone therapy for AA in the largest reported cohort of patients; to identify patient and treatment factors predictive of therapeutic success; and to develop a practical model for predicting patient response. METHODS: The medical records of 148 consecutive patients treated with diphencyprone were reviewed. A clinically significant response to diphencyprone therapy was defined as a cosmetically acceptable response or greater than 75% terminal hair regrowth. Survival analyses using the Kaplan-Meier method and the Cox proportional hazards model were performed to determine significant factors predictive of regrowth and relapse. RESULTS: Using a survival analysis model, the cumulative patient response at 32 months was 77.9% (95% confidence interval, 56.8%-98.9%). Variables independently associated with clinically significant regrowth were age at onset of disease and baseline extent of AA. Older age at onset of AA portended a better prognosis. A cosmetically acceptable end point was obtained in 17.4% of patients with alopecia totalis/universalis, 60.3% with 75% to 99% AA, 88.1% with 50% to 74% AA, and 100% with 25% to 49% AA. A lag of 3 months was present between initiation of therapy and development of significant hair regrowth in the first responders. Relapse after achieving significant regrowth developed in 62.6% of patients. CONCLUSIONS: Response to diphencyprone treatment in AA is affected by baseline extent of AA and age at disease onset. A prolonged treatment course might be necessary. A predictive model has been developed to assist with patient prognostication and counseling. PMID- 11493100 TI - Complete remission of scleromyxedema following autologous stem cell transplantation. PMID- 11493101 TI - A better potassium hydroxide preparation? In vivo diagnosis of tinea with confocal microscopy. AB - BACKGROUND: Traditional diagnostic testing for dermatophyte infection currently requires skin scraping for light microscopy and/or fungal culture or skin biopsy. Immunofluorescent microscopy can also be used with calcofluor stain. All of these tests can be time-consuming to perform, require a waiting period for results, and are invasive. We investigated the use of a real-time, noninvasive, confocal microscope in visualizing dermatophyte hyphae in vivo. OBSERVATIONS: Confocal microscopic imaging of active tinea can clearly identify dermatophyte hyphae within the upper epidermis after potassium hydroxide application. The hyphae appear as bright linear branching objects not found in uninvolved skin. CONCLUSIONS: It is possible to immediately and painlessly image dermatophyte hyphae in active lesions of tinea by means of a confocal microscope. With further improvement, imaging devices may be available to physicians to instantly and noninvasively evaluate a variety of skin disorders in microscopic detail. PMID- 11493102 TI - Does the tail wag the dog? Role of the barrier in the pathogenesis of inflammatory dermatoses and therapeutic implications. PMID- 11493104 TI - Defined UV protection by apparel textiles. AB - This article was written to update information on test methods and standards for determining the UV protection of apparel textiles and on factors affecting UV protective properties of fabrics, from dermatological and textile technological viewpoints. Articles from dermatological and textile technological journals published from 1990 to 2001 were identified from MEDLINE, Excerpta Medica/EMBASE, World Textiles, and Textile Technology Digest. Peer-reviewed dermatological articles, textile technological research articles, and normative publications were selected. Independent data extraction was performed by several observers. Spectrophotometry is the preferred method for determining UV protection factor of textile materials. Various textile qualities affect the UV protection factor of a finished garment; important elements are the fabric porosity, type, color, weight, and thickness. The application of UV absorbers in the yarns significantly improves the UV protection factor of a garment. With wear and use, several factors can alter the UV protective properties of a textile, including stretch, wetness, and degradation due to laundering. Standards in the field exist in Australia and Great Britain, and organizations such as the European Standardization Commission in Europe and the American Association of Textile Chemists and Colorists and the American Society for Testing and Materials in the United States are also establishing standards for the determination and labeling of sun protective clothing. Various textile qualities and conditions of wear and use affect UV protective properties of apparel textiles. The use of UV blocking fabrics can provide excellent protection against the hazards of sunlight; this is especially true for garments manufactured as UV protective clothing. PMID- 11493105 TI - Off-center fold: crusted erythematous postauricular plaques. PMID- 11493106 TI - Off-center fold: a congenital midline nasal mass in a newborn. PMID- 11493107 TI - Off-center fold: verrucous penile lesions on a man with paraplegia. PMID- 11493108 TI - Off-center fold: linear atrophy, telangiectases, and soft nodules along the lines of Blaschko with skeletal abnormalities. PMID- 11493109 TI - Methodological limitations of the study "Isotretinoin use and risk of depression, psychotic symptoms, suicide, and attempted suicide". PMID- 11493111 TI - Cutaneous warts in HIV-positive patients undergoing highly active antiretroviral therapy. PMID- 11493112 TI - Pancytopenia after treatment of keratoacanthoma by single lesional methotrexate infiltration. PMID- 11493113 TI - Continue reading. PMID- 11493114 TI - Skin preparation for laser therapy. PMID- 11493115 TI - Photoinduced Sweet syndrome. PMID- 11493116 TI - Chronic penile lymphedema: a report of 6 cases. PMID- 11493117 TI - Ceramide-dominant, barrier-repair lipids improve childhood atopic dermatitis. PMID- 11493121 TI - Religion and medicine. PMID- 11493122 TI - Long-term pharmacotherapy of obesity 2000: a review of efficacy and safety. AB - To clarify the efficacy of antiobesity drugs, this article reviews all long-term (> or =36 weeks), placebo-controlled trials of obesity pharmacotherapy published since 1960. Since fears of anorexiant-induced heart valve damage preclude many physicians and patients from even considering antiobesity drugs, this area is also reviewed in-depth. Electronic database and manual bibliography search was used to identify all relevant publications. While existing studies are too few and heterogeneous to warrant meta-analysis, their review does provide evidence highly relevant to the safety and efficacy of available anorexiants. Weight loss attributable to obesity pharmacotherapy (ie, in excess of placebo) in trials lasting 36 to 52 weeks was 8.1% or 7.9 kg for those receiving phentermine resin, 5.0 % or 4.3 kg for those receiving sibutramine hydrochloride, 3.4% or 3.4 kg for those receiving orlistat, and -1.5% or -1.5 kg for those receiving diethylpropion hydrochloride. Physiologic, pathologic, and epidemiological studies strongly support that anorexiant-induced valvulopathy is attributable to specific serotonergic properties of the fenfluramines that are not present with available weight loss drugs. PMID- 11493123 TI - Evaluation of patients with chest pain and normal coronary angiograms. PMID- 11493124 TI - Monotherapy may be suboptimal for severe bacteremic pneumococcal pneumonia. AB - BACKGROUND: Although monotherapy for pneumococcal pneumonia is standard, a survival benefit of combination beta-lactam and macrolide therapy has been suggested. HYPOTHESIS: Initial empirical therapy with a combination of effective antibiotic agents would have a better outcome than a single effective antibiotic agent in patients with bacteremic pneumococcal pneumonia. METHODS: A review of adult bacteremic pneumococcal pneumonia within the Methodist Healthcare System, Memphis, Tenn, between January 1, 1996, and July 31, 2000. Empirical therapy was defined as all antibiotic agents received in the first 24 hours after presentation. On the basis of culture results, empirical therapy was classified as single effective therapy (SET), dual effective therapy (DET), or more than DET (MET). Acute Physiology and Chronic Health Evaluation II (APACHE II)-based predicted mortality, and Pneumonia Severity Index scores were calculated. RESULTS: Of the 225 patients identified, 99 were classified as receiving SET, 102 as receiving DET, and 24 as receiving MET. Compared with the other groups, patients who received MET had statistically significantly more severe pneumonia as measured by the Pneumonia Severity Index score (P =.04) and predicted mortality (P =.03). Mortality within the SET group was significantly higher than within the DET group (P =.02, odds ratio, 3.0 [95% confidence intervals, 1.2 7.6]), even when the DET and MET groups (P =.04) were combined. In a logistic regression model including antibiotic therapy and clinical risk factors for mortality, SET remained an independent predictor of mortality with a predicted mortality-adjusted odds ratio for death of 6.4 (95% confidence intervals, 1.9 21.7). All deaths occurred in patients with a Pneumonia Severity Index score higher than 90, and the predicted mortality-adjusted odds ratio for death with SET in this subgroup was 5.5 (95% confidence intervals, 1.7-17.5). CONCLUSIONS: We found that SET is associated with a significantly greater risk of death than DET. Therefore, monotherapy may be suboptimal for patients with severe bacteremic pneumococcal pneumonia who have Pneumonia Severity Index scores higher than 90. PMID- 11493125 TI - Better psychological functioning and higher social status may largely explain the apparent health benefits of wine: a study of wine and beer drinking in young Danish adults. AB - BACKGROUND: Findings from a recent series of Danish studies suggest that moderate wine drinkers are healthier than those who drink other alcoholic beverages or those who abstain. OBJECTIVE: To identify possible explanatory factors associated with the health benefits of wine consumption through the examination of a wide spectrum of social, cognitive, and personality characteristics related to both beverage choice and health in young Danish adults. SUBJECTS AND METHODS: Descriptive cross-sectional study of characteristics associated with beverage choice in a sample of 363 men and 330 women between the ages of 29 and 34 years, selected from the Copenhagen Perinatal Cohort on the basis of perinatal records. MAIN OUTCOME MEASURES: Socioeconomic status, education, IQ, personality, psychiatric symptoms, and health-related behaviors, including alcohol consumption, were analyzed. The outcome variables were subjected to linear and logistic regression analyses with 2 factors (beer and wine), each with 2 levels (drinking or not drinking a certain beverage type). RESULTS: Wine drinking was significantly associated with higher IQ, higher parental educational level, and higher socioeconomic status. Beer drinking was significantly associated with lower scores on the same variables. On scales concerning personality, psychiatric symptoms, and health-related behaviors, wine drinking was associated with optimal functioning and beer drinking with suboptimal functioning. CONCLUSIONS: Our data demonstrate that wine drinking is a general indicator of optimal social, cognitive, and personality development in Denmark. Similar social, cognitive, and personality factors have also been associated with better health in many populations. Consequently, the association between drinking habits and social and psychological characteristics, in large part, may explain the apparent health benefits of wine. PMID- 11493127 TI - Dietary patterns and the risk of coronary heart disease in women. AB - BACKGROUND: Although substantial information on individual nutrients or foods and risk of coronary heart disease (CHD) is available, little is known about the role of overall eating pattern. METHODS: Using dietary information from a food frequency questionnaire in 1984 from the Nurses' Health Study, we conducted factor analysis and identified 2 major dietary patterns-"prudent" and "Western" and calculated factor scores of each pattern for individuals in the cohort. We used logistic regression to examine prospectively the associations between dietary patterns and CHD risk among 69 017 women aged 38 to 63 years in 1984 without history of major chronic diseases. RESULTS: The prudent pattern was characterized by higher intakes of fruits, vegetables, legumes, fish, poultry, and whole grains, while the Western pattern was characterized by higher intakes of red and processed meats, sweets and desserts, french fries, and refined grains. Between 1984 and 1996, we documented 821 CHD cases. After adjusting for coronary risk factors, the prudent diet score was associated with a relative risk (RR) of 0.76 (95% confidence interval (CI), 0.60-0.98; P for trend test,.03) comparing the highest with lowest quintile. Extreme quintile comparison yielded an RR of 1.46 (95% CI, 1.07-1.99; P for trend test,.02) for the Western pattern. Those who were jointly in the highest prudent diet quintile and lowest Western diet quintile had an RR of 0.64 (95% CI, 0.44-0.92) compared with those with the opposite pattern profile. CONCLUSION: A diet high in fruits, vegetables, whole grains, legumes, poultry, and fish and low in refined grains, potatoes, and red and processed meats may lower risk of CHD. PMID- 11493126 TI - Relationship of depression to increased risk of mortality and rehospitalization in patients with congestive heart failure. AB - BACKGROUND: Patients with congestive heart failure (CHF) may have a high prevalence of depression, which may increase the risk of adverse outcomes. OBJECTIVE: To determine the prevalence and relationship of depression to outcomes of patients hospitalized with CHF. METHODS: We screened patients aged 18 years or older having New York Heart Association class II or greater CHF, an ejection fraction of 35% or less, or both, admitted between March 1, 1997, and June 30, 1998, to the cardiology service of one hospital. Patients with a Beck Depression Inventory score of 10 or higher underwent a modified National Institute of Mental Health Diagnostic Interview Schedule to identify major depressive disorder. Primary care providers coordinated standard treatment for CHF and other medical and psychiatric disorders. We assessed all-cause mortality and readmission (rehospitalization) rates 3 months and 1 year after depression assessment. Logistic regression analyses were used to evaluate the independent prognostic value of depression after adjustment for clinical risk factors. RESULTS: Of 374 patients screened, 35.3% had a Beck Depression Inventory score of 10 or higher and 13.9% had major depressive disorder. Overall mortality was 7.9% at 3 months and 16.2% at 1 year. Major depression was associated with increased mortality at 3 months (odds ratio, 2.5 vs no depression; P =.08) and at 1 year (odds ratio, 2.23; P =.04) and readmission at 3 months (odds ratio, 1.90; P =.04) and at 1 year (odds ratio, 3.07; P =.005). These increased risks were independent of age, New York Heart Association class, baseline ejection fraction, and ischemic etiology of CHF. CONCLUSIONS: Major depression is common in patients hospitalized with CHF and is independently associated with a poor prognosis. PMID- 11493128 TI - Nonsevere community-acquired pneumonia: correlation between cause and severity or comorbidity. AB - BACKGROUND: Community-acquired pneumonia frequently constitutes a nonsevere infection manageable at home. However, for these low-risk episodes, the epidemiological features have not been carefully analyzed. OBJECTIVES: To determine the cause of nonsevere community-acquired pneumonia and to investigate if a correlation exists between cause and severity or comorbidity. METHODS: During a 3-year period, all patients with nonsevere community-acquired pneumonia, according to the Pneumonia Patient Outcome Research Team prognostic classification (patients in groups 1-3), were included in the study. Causes were investigated through the following procedures: cultures of blood, sputum, and pleural fluid; serologic tests; and polymerase chain reaction methods to detect Streptococcus pneumoniae DNA in whole blood or Mycoplasma pneumoniae and Chlamydia pneumoniae DNA in throat swab specimens. RESULTS: Of 317 initially included patients, 247 were eligible for the study. A microbial diagnosis was obtained in 162 patients (66%), and the main pathogens detected were S pneumoniae (69 patients [28%]), M pneumoniae (40 patients [16%]), and C pneumoniae (28 patients [11%]). For the 58 patients in prognostic group 1, M pneumoniae was the most prevalent cause, and atypical microorganisms constituted 40 (69%) of the isolated agents. In contrast, for patients in prognostic groups 2 and 3, S pneumoniae was the leading agent, and a significant reduction of M pneumoniae cases and a greater presence of other more uncommon pathogens were observed. The existence of comorbid conditions was not a determining factor for particular causes. CONCLUSIONS: Among low-risk patients with community-acquired pneumonia, there was a certain correlation between severity and cause. In contrast, the existence of a comorbidity did not have a predictive causative value. PMID- 11493129 TI - Antihypertensive drug therapy in Saskatchewan: patterns of use and determinants in hypertension. AB - BACKGROUND: The benefits of continuous treatment of hypertension have been extensively documented in randomized controlled trials. However, clinical trials may not reflect actual drug use in the population. OBJECTIVE: To examine the distribution and determinants of patterns of use of antihypertensive agents in the first 5 years of hypertension treatment in Saskatchewan. METHODS: Patterns of use and modifications to therapy were derived from a careful examination of medication use in a cohort of 19 501 subjects aged 40 to 79 years, without recognized cardiac disease and initiating therapy with an angiotensin-converting enzyme inhibitor, a calcium antagonist, or a beta-blocker in Saskatchewan between 1990 and 1993. RESULTS: Angiotensin-converting enzyme inhibitors (37.4%), followed by calcium antagonists (27.5%) and beta-blockers (26.4%), were the most commonly prescribed agents to initiate treatment in our study population. Patients with diabetes were less likely to be dispensed a beta-blocker, as were younger and female patients. Previous visits to a cardiologist decreased the likelihood of receiving combination therapy or angiotensin-converting enzyme inhibitors but increased that of using calcium antagonists. Apart from dose adjustment, 89% of study subjects underwent at least 1 modification to their initial regimen, at a median time of 134 days. After 1 year, only 33.8% of patients were still using their initial drug. An early decrease in the proportion of patients continuing to receive initial therapy was noted, especially among beta-blocker users. CONCLUSIONS: Erratic drug-taking behaviors were observed in this Saskatchewan population. In addition, initial drug use does not seem to be in accordance with the stepped-care approach to hypertension therapy recommended in the Canadian guidelines. PMID- 11493130 TI - Religious struggle as a predictor of mortality among medically ill elderly patients: a 2-year longitudinal study. AB - BACKGROUND: Although church attendance has been associated with a reduced risk of mortality, no study has examined the impact of religious struggle with an illness on mortality. OBJECTIVE: To investigate longitudinally the relationship between religious struggle with an illness and mortality. METHODS: A longitudinal cohort study from 1996 to 1997 was conducted to assess positive religious coping and religious struggle, and demographic, physical health, and mental health measures at baseline as control variables. Mortality during the 2-year period was the main outcome measure. Participants were 596 patients aged 55 years or older on the medical inpatient services of Duke University Medical Center or the Durham Veterans Affairs Medical Center, Durham, NC. RESULTS: After controlling for the demographic, physical health, and mental health variables, higher religious struggle scores at baseline were predictive of greater risk of mortality (risk ratio [RR] for death, 1.06; 95% confidence interval [CI], 1.01-1.11; chi(2) = 5.89; P =.02). Two spiritual discontent items and 1 demonic reappraisal item from the religious coping measure were predictive of increased risk for mortality: "Wondered whether God had abandoned me" (RR for death, 1.28; 95% CI, 1.07-1.50; chi(2) = 5.22; P =.02), "Questioned God's love for me" (RR for death, 1.22; 95% CI, 1.02-1.43; chi(2) = 3.69; P =.05), and "Decided the devil made this happen" (RR for death, 1.19; 95% CI, 1.05-1.33; chi(2) = 5.84; P =.02). CONCLUSIONS: Certain forms of religiousness may increase the risk of death. Elderly ill men and women who experience a religious struggle with their illness appear to be at increased risk of death, even after controlling for baseline health, mental health status, and demographic factors. PMID- 11493131 TI - Diagnostic patterns and temporal trends in the evaluation of adult patients hospitalized with syncope. AB - BACKGROUND: Syncope is a common clinical problem that is often difficult and expensive to diagnose. We examined diagnostic patterns and trends and use of specialty consultations in the evaluation of syncope. METHODS: We retrospectively reviewed the medical records of consecutive adult patients hospitalized with the principal diagnosis of syncope (International Classification of Diseases, Ninth Revision, code 780.2) during 1994 and 1998 at 2 community teaching hospitals. RESULTS: A total of 649 patients (57% female) with a mean (+/-SD) age of 68 +/- 15 years were identified in 1994 (n = 451) and 1998 (n = 198). Three hundred forty-one patients (53%) underwent at least 1 neurologic test, including brain computed tomographic (CT) scan (n = 283), electroencephalography (n = 253), carotid Doppler echocardiography (n = 185), and brain magnetic resonance imaging (n = 10). Only brain CT scan and electroencephalography yielded diagnoses in 5 (2%) and 6 patients (2%), respectively with history consistent with seizures or stroke. Cardiovascular tests providing the highest diagnostic yields (postural blood pressure check in 52 [30%], head-up tilt-table test in 32 [24%], and electrophysiologic study in 5 [16%]) were used in 176 (27%), 132 (20%), and 31 patients (5%), respectively. Differences in the use of some tests were noted at the participating hospitals and over time (1994 vs 1998). The total number of diagnosed cases was similar for patients undergoing evaluation by primary care physicians alone (65/103 [63%]), compared with cardiology (48/85 [56%]), neurology (29/48 [60%]), or both (81/141 [57%]). After a mean (+/-SD) length of stay of 5 +/- 4 days, 320 (49%) of 649 cases remained undiagnosed. CONCLUSIONS: Despite a reduction in the use of some tests (eg, brain CT scan and carotid Doppler) over time, lower-yield neurologic tests were overused and higher-yield cardiovascular tests were likely underused. The untargeted, seemingly random use of specialty evaluations did not seem to contribute to an increase in the overall number of diagnosed cases. Increased use of specific tests directed by history and results of physical examination may improve diagnostic yield and decrease the cost of evaluating syncope. PMID- 11493132 TI - Academic detailing to improve use of broad-spectrum antibiotics at an academic medical center. AB - BACKGROUND: Antibiotic misuse is common and costly and may promote antibiotic resistance. We tested the efficacy of a targeted one-on-one educational program ("academic detailing") designed to improve the appropriateness of broad-spectrum antibiotic use. METHODS: A randomized controlled trial was conducted in a large US teaching hospital. During an 18-week study period, 17 general medical, oncology, and cardiology services either received academic detailing or did not. The intervention was prompted by an order for either levofloxacin or ceftazidime that led to a computer-based review of data for that patient. Orders for the 2 target antibiotics deemed unnecessary by a priori criteria were included in the study. The primary outcome examined was the number of days that unnecessary levofloxacin or ceftazidime was administered in intervention and control groups. RESULTS: Before the trial, intervention and control services had similar prescribing patterns for the target antibiotics; the drugs were used for similar indications throughout the study period. During the intervention, there was a reduction of 37% in days of unnecessary levofloxacin or ceftazidime use per 2 week interval on services randomized to the educational intervention vs control services (P< .001). In multivariable analyses controlling for baseline prescribing and study interval, the rate of unnecessary use of the 2 target antibiotics was reduced by 41% on the intervention services compared with controls (95% confidence interval, 44%-78%; P< .001). Length of stay, intensive care unit transfers, readmission rates, and in-hospital death rates were similar in both groups (P> or =.10 for all). CONCLUSION: Targeted one-on-one education is a practical, effective, and safe method for reducing excessive broad-spectrum antibiotic use. PMID- 11493133 TI - Relationship between systemic markers of inflammation and serum beta-carotene levels. AB - BACKGROUND: Low serum levels of beta-carotene have been associated with increased risk of cancer and cardiovascular disease. However, in clinical trials, supplementation of the diet with beta-carotene either had no benefit or caused harm. This pattern of findings raises the possibility that confounding by other factors might explain the association between serum beta-carotene level and disease risk. METHODS: We used data from 14 470 current smokers, ex-smokers, and never smokers aged 18 years or older who participated in the Third National Health and Nutrition Examination Survey to assess the relationship between serum beta-carotene and markers of inflammation (C-reactive protein and white blood cell count). RESULTS: After adjustment for beta-carotene intake and other factors, geometric mean levels of serum beta-carotene for individuals with undetectable (< 0.22 mg/dL), mildly elevated (0.22-0.99 mg/dL), and clinically elevated (> or =1.0 mg/dL) C-reactive protein levels were 18.0, 16.1, and 13.6 microg/dL, respectively, in never smokers; 18.1, 15.7, and 13.9 microg/dL in ex smokers; and 11.3, 10.2, and 9.4 microg/dL in current smokers (P< .001 for all). In corresponding analyses, white blood cell count was also inversely related to serum beta-carotene concentration (P< .05 for all). CONCLUSIONS: The strong and inverse association of serum beta-carotene level with C-reactive protein level and white blood cell count suggests that the relationship between serum beta carotene concentration and disease risk might be confounded by inflammation. More broadly, for beta-carotene and likely other nutrients, it seems unwise to interpret biomarker data as prima facie evidence of dietary intake without a more complete understanding of the physiologic processes that affect nutrient levels. PMID- 11493134 TI - Possible relationship between statin use and decreased incidence of dementia: are we ready for a new indication for these drugs? PMID- 11493135 TI - Pacemaker infection due to Brucella melitensis: a case report. PMID- 11493136 TI - Bone marrow failure from medication error: diagnosis by history, not biopsy. PMID- 11493137 TI - Amelanotic melanoma presenting as a scar. PMID- 11493138 TI - Truly universal access. PMID- 11493139 TI - Steps toward universal health care. PMID- 11493140 TI - A collaborative responsibility for health. PMID- 11493141 TI - Cost-saving features of universal coverage. PMID- 11493142 TI - Components of a rational and workable health care system. PMID- 11493144 TI - Thoughtful discharge planning for better outcomes. PMID- 11493146 TI - Prospective payment system and hospitalization for pneumonia in Italy. PMID- 11493148 TI - Tea and coronary heart disease: the flavonoid quercetin is more bioavailable from rutin in women than in men. PMID- 11493149 TI - Drug combination for asthma treatment. PMID- 11493151 TI - Systemic thromboembolism in atrial fibrillation. PMID- 11493153 TI - The case for compensation of tissue donors. PMID- 11493155 TI - Is smoking a risk factor for pulmonary embolism? PMID- 11493157 TI - The diagnosis of imported malaria. PMID- 11493158 TI - Peripheral neuropathy as the first clinical manifestation of Wegener granulomatosis. PMID- 11493159 TI - Subthalamic deep brain stimulation: neurobehavioral concerns. PMID- 11493160 TI - The role of hyperglycemia in acute stroke. AB - BACKGROUND: Ischemic stroke is a leading cause of death and long-term disability, and hyperglycemia is believed to aggravate cerebral ischemia. OBJECTIVES: To review animal and human studies on the relationship between hyperglycemia and brain ischemia that elucidate some of the mechanisms for the deleterious effect of hyperglycemia. To discuss present and future clinical recommendations for glucose control. METHODS: Computerized data sources and published indexes and articles from 1976 through 2000 were searched for human studies that evaluated the association between stroke and hyperglycemia, and studies focused on experimental models of hyperglycemic animals with focal and global brain ischemia. RESULTS: Most human studies have shown that in acute stroke, admission hyperglycemia in patients with or without diabetes is associated with a worse clinical outcome than in patients without hyperglycemia. This association is more consistent in the nonlacunar type of stroke. Animal studies support these findings by showing both in global and in focal postischemic models that hyperglycemia exaggerates the following damaging processes: intracellular acidosis, accumulation of extracellular glutamate, brain edema formation, blood brain barrier disruption, and tendency for hemorrhagic transformation. Insulin treatment of hyperglycemic animals was found to have a beneficial effect in focal and global brain ischemia, which may be mediated by the glucose-reduction effect or by a direct neuroprotection. CONCLUSIONS: Most studies show the deleterious effect of early hyperglycemia, especially in patients with nonlacunar focal or global ischemia. Clinical trials of intensive insulin treatment are needed. Meanwhile simple measures to avoid excessive hyperglycemia are recommended. PMID- 11493161 TI - Standardized neurologic evaluations of 128 patients with Wegener granulomatosis. AB - OBJECTIVE: To assess the frequency and type of neurologic involvement in a cohort of patients with generalized Wegener granulomatosis (WG). PATIENTS AND METHODS: In a prospective analysis the clinical, electrophysiologic, radiological, and serologic data of 128 patients have been studied over a median observation period of 19 months (range, 1-60 months). RESULTS: Sixty-four patients (50%) revealed central or peripheral nervous system involvement. Peripheral neuropathy (PN) affected 56 patients, in 9 cases the central nervous system was involved, and in 6 cases the cranial nerves were involved. Thirty-one patients showed a distal symmetrical polyneuropathy, 25 a mononeuritis multiplex. Within the first 2 years of the disease course 47 of the 56 patients had developed their PN, sometimes as the initial symptom of WG. Patients with PN were significantly more often male (34 of 65 patients) than female (22 of 63 patients, P =.04), were significantly older at the onset of WG (median age, 53 vs 44 years; P =.001), had a significantly larger disease extent (P =.001), and had higher classic antineutrophil cytoplasmic antibody titers (P =.002) than neurologically unaffected patients. Response to immunosuppression was moderate concerning peripheral nervous system manifestations. CONCLUSIONS: Peripheral neuropathy is frequent in generalized WG, occurring early in the disease course. As PN can be the first and sole symptom of a beginning systemic vasculitis, it is important that in cases of PN of an unclear origin, interdisciplinary investigations are initiated to detect, treat, and closely follow-up a possible underlying WG, especially as these patients seem to have a more severe disease course. PMID- 11493162 TI - Effects of bilateral subthalamic stimulation on cognitive function in Parkinson disease. AB - BACKGROUND: Chronic bilateral subthalamic deep brain stimulation (STN-DBS) is known to improve motor function in patients with Parkinson disease (PD). However, the possible effects of STN-DBS on neuropsychological functions have been studied less. OBJECTIVE: To investigate the effects of STN-DBS on neuropsychological functions in PD. DESIGN: Before-after trial. PATIENTS AND METHODS: Fifteen consecutive patients were assessed before and 3 months after implantation of stimulators for STN-DBS (postsurgical assessment with the stimulators switched on). Both assessments were performed with patients in a drug-free condition. The neuropsychological battery consisted of tests measuring memory and visuospatial and frontal functions. RESULTS: The comparison between presurgical and postsurgical performance showed a moderate deterioration in verbal memory and prefrontal and visuospatial functions, and a moderate improvement in a prefrontal task and obsessive-compulsive traits. The motor state improved in all patients. CONCLUSION: Therapy with STN-DBS improves motor symptoms in PD without any clinically relevant neuropsychological deterioration. PMID- 11493163 TI - Contribution of asymmetric synapse loss to lateralizing clinical deficits in frontotemporal dementias. AB - BACKGROUND: Synapse loss has been found to be the major correlate of cognitive decline in Alzheimer disease (AD), and prefrontal synapse loss has been found in patients with frontotemporal dementia (FTD). OBJECTIVE: To see if our hypothesis that within each FTD case, regional synapse loss would correlate with lateralizing neuropsychologic and neurobehavioral deficits would be correct. DESIGN: We analyzed synaptophysin as a marker for synapse loss in snap-frozen brain samples, using an enzyme-linked immunosorbent assay technique. Quantitative results were obtained by comparing patient data with a standard curve made by analyzing serial dilutions of a recombinant synaptophysin protein fragment. A board-certified neuropsychologist and a board-certified neurologist, both unaware of the synaptophysin results, determined areas of primary neuropsychologic and neurobehavioral dysfunction. Relationships between areas of primary dysfunction and synapse loss were analyzed using the binomial test. PATIENTS: Six cases of FTD, 28 cases of AD, and 16 nondemented control subjects. RESULTS: Five of 6 FTD cases had regional synaptophysins correlating with lateralizing frontal or temporal deficits. Three of 6 correlated in 2 or more regions. Although our results were higher than that expected based on a pure-chance mechanism (50% vs 34%), statistical significance was not attained. CONCLUSIONS: We found a trend for an association between synapse loss and lateralizing neuropsychologic and neurobehavioral deficits in FTD. Studies in larger numbers of FTD cases are planned with the goal of attaining statistically significant conclusions. PMID- 11493165 TI - Inclusion body myositis mimicking motor neuron disease. AB - OBJECTIVE: To describe the clinical and electrophysiologic features of patients with inclusion body myositis that was misinterpreted as motor neuron disease. PATIENTS AND METHODS: We retrospectively retrieved the medical records of 70 patients with a pathologic diagnosis of inclusion body myositis. From this group, we selected those who had been first diagnosed as having motor neuron disease or amyotrophic lateral sclerosis. We reviewed the clinical, electrophysiologic, laboratory, and morphologic studies. RESULTS: Nine (13%) of 70 patients with inclusion body myositis had been diagnosed as having motor neuron disease. Six of the 9 patients had asymmetric weakness; in 4 the distal arm muscles were affected. Eight patients had finger flexor weakness. Tendon reflexes were preserved in weak limbs in 6, hyperactive in 2, and absent in 1. Four patients had dysphagia. Fasciculation was seen in 2 patients. None had definite upper motor neuron signs or muscle cramps. Routine electromyographic studies showed fibrillation potentials and positive sharp waves in all 9. Fasciculation potentials were seen in 7 and long-duration polyphasic motor unit potentials were seen in 8. There was no evidence of a myogenic disorder in these 9 patients. Muscle biopsy was done because of slow progression or prominent weakness of the finger flexors and was diagnostic of inclusion body myositis. A quantitative electromyogram was myopathic in 4 of the 5 patients studied. CONCLUSIONS: Inclusion body myositis may mimic motor neuron disease. Muscle biopsy and quantitative electromyographic analysis are indicated in patients with atypical motor neuron disease, especially those with slow progression or early and disproportionate weakness of the finger flexors. PMID- 11493164 TI - The effects of age on rate of progression of Alzheimer disease and dementia with associated cerebrovascular disease. AB - BACKGROUND: Relatively little is known about how cerebrovascular disease affects progression of dementia. Previous studies have found no differences in progression of Alzheimer disease and vascular dementia, but these studies have not specifically examined age effects. OBJECTIVE: To test whether the rate of cognitive decline is different in Alzheimer disease compared with dementia with associated cerebrovascular disease in clinical and autopsy patient series. PATIENTS AND METHODS: We studied the longitudinal course of cognitive function as measured by the Mini-Mental State Examination (MMSE) in patients with clinically and neuropathologically diagnosed conditions evaluated through a university Alzheimer disease center. Clinical patients were grouped according to possible Alzheimer disease without stroke (n = 37), probable Alzheimer disease without stroke (n = 181), and dementia with stroke (n = 50). Autopsy cases were categorized into Alzheimer disease (n = 78) and dementia with vascular disease (n = 13). Data were analyzed using random-effects modeling of longitudinal change. RESULTS: There was a significant interaction between age and diagnosis in determining rate of change on the MMSE scores for both the clinical and autopsy samples. Rate of change decreased slightly with advancing age for Alzheimer disease groups, but increased with age for dementia with cerebrovascular disease groups. CONCLUSIONS: Dementia with cerebrovascular disease declined faster in patients 80 years and older compared with Alzheimer disease without associated cerebrovascular pathological conditions, but showed slower decline in patients younger than 80 years. This effect most likely reflects combined Alzheimer and vascular pathological conditions in older patients with cerebrovascular disease. PMID- 11493166 TI - Brain single-photon emission computed tomography and magnetic resonance imaging in Machado-Joseph disease. AB - BACKGROUND: Machado-Joseph disease (MJD) is one of the most frequently encountered spinocerebellar ataxias. However, few reports on brain single-photon emission computed tomographic (SPECT) imaging (BSI) with hexylmethylpropylene amineoxine labled with technetium Tc 99m and magnetic resonance imaging (MRI) have been performed for the evaluation of patients with MJD. OBJECTIVES: To investigate possible abnormalities with BSI and MRI in patients with MJD and to correlate these findings with the duration of symptoms; cerebellar, extrapyramidal, and pyramidal syndromes; and the molecular characteristics of the MJD mutation. PATIENTS AND METHODS: Twelve patients (8 males and 4 females [mean age, 39 years]) with genetically proven MJD were studied. The patients underwent BSI and MRI on the same day. Brain SPECT imaging was performed after an intravenous injection of 99mTc-hexylmethylpropylene amineoxine. The transaxial, coronal, and sagittal BSIs obtained were submitted to visual and semiquantitative analyses. Magnetic resonance imaging was obtained in a 2-T system with coronal, sagittal, transaxial, and 3-dimensional (volumetric) acquisitions. The volumes of the cerebellar hemispheres and vermis were calculated. Control groups for BSI (22 female and 20 male subjects [mean age, 33 years]) and MRI (13 female and 4 male subjects [mean age, 32.2 years]) were included for comparison. RESULTS: Correlation was observed between the perfusion abnormalities identified by visual analysis in the BSI with the structural abnormalities observed on MRI in the parietal lobes and vermis. Brain SPECT imaging identified (by visual analysis) more perfusion abnormalities in the inferior portion of the frontal lobes, mesial and lateral portions of the temporal lobes, basal ganglia, and cerebellar hemispheres. Magnetic resonance imaging identified more abnormalities in the pons and superior portions of the frontal lobes. Olivary atrophy was identified by MRI. Semiquantitative analysis showed a statistically significant difference of perfusion in the inferior and superior portions of the frontal lobes, lateral portion of the temporal lobes, parietal lobes, left basal ganglia, cerebellar hemispheres, and vermis when compared with the control group. A significant difference was noted between the vermis and cerebellar volumes on MRI when compared with the control group. A significant relationship was observed between the perfusion of the left parietal lobe (P =.05) and extrapyramidal syndrome. There was a tendency toward an inverse relationship between the duration of symptoms and the perfusion of the cerebellar hemispheres (rho = -0.37; P =.24) and volume of the vermis (rho = -0.30; P =.34); between the length of the expanded (CAG)n repeat and the perfusion of the left parietal lobe (rho = -0.32; P =.36), vermis (rho = -0.28; P =.43), and pons (rho = -0.28; P =.42). A direct association was observed between the length of the expanded (CAG)n repeat and the perfusion of the lateral portion of the right temporal lobe (rho = 0.67; P =.03). CONCLUSIONS: Brain SPECT imaging and MRI were capable of identifying subclinical abnormalities in individuals with MJD. These findings may be helpful for a better understanding of the pathophysiology of this disease. PMID- 11493167 TI - Effect of divalproex-lamotrigine combination therapy in frontal lobe seizures. AB - CONTEXT: Patients with intractable frontal lobe seizures represent a difficult subclass of patients with epilepsy. When medications fail, surgical outcomes typically have not been as successful as medial temporal lobe resections. The combination therapy of valproic acid (divalproex sodium) and lamotrigine has shown promising results in patients with uncontrolled seizures. OBJECTIVE: To determine outcome in patients with intractable frontal lobe seizures who were treated with the combination of divalproex and lamotrigine. DESIGN: A nonrandomized, open-label, add-on trial. SETTING: Outpatients evaluated and treated at a tertiary care referral facility. PATIENTS: Twenty-one patients between 16 and 65 years old were studied. Patients were required to have failed at least 3 prior trials with antiepilepsy drugs. Criteria for frontal lobe onset included 1 or more of the following: frontal lesion on scan, positive ictal single-photon emission computed tomographic scan, symptoms consistent with frontal lobe onset, or an electroencephalogram (surface or invasive) consistent with frontal lobe onset. INTERVENTION: Patients were treated with divalproex lamotrigine combination therapy for 1 year. MAIN OUTCOME MEASURES: The main outcome measured was seizure reduction. Safety and tolerability were also evaluated. RESULTS: Four patients discontinued therapy. Ten of the remaining 17 became completely free of seizures. Two rashes occurred, but did not lead to discontinuation of therapy. The most common adverse events were tremor and weight gain. CONCLUSION: Divalproex-lamotrigine combination therapy is a reasonable alternative in intractable frontal lobe epilepsy. PMID- 11493168 TI - Moyamoya: Indiana University Medical Center experience. AB - BACKGROUND: Moyamoya usually presents with cerebral ischemia in children and intracranial hemorrhage in adults. Treatment remains controversial. DESIGN AND OBJECTIVE: We reviewed our experience from June 1995 to August 1999 of 20 adult and pediatric angiographically diagnosed patients with moyamoya to report their clinical presentation, radiological findings, management, and clinical outcomes. RESULTS: The mean age of patients at symptom onset was 17 years (range, 2-54 years). Patients were divided into 2 age groups (group 1, <18 years; group 2, > or =18 years). There were 13 patients in group 1 and 7 patients in group 2. Ischemic strokes or transient ischemic attacks were the predominant initial presentations in both groups. One patient in group 2 had an intraparenchymal brain hemorrhage. Five patients received medical treatment, and 15 had surgical revascularization. The mean time from symptom onset to surgical procedure was significantly longer for patients in group 1 than for those in group 2 (P =.03). The mean follow-up time was 36 months. One patient in group 1 had an ischemic stroke. There was no difference in stroke recurrence, mortality, or modified Rankin scale score among medically or surgically treated patients. CONCLUSIONS: Moyamoya disease may have a different presentation and more benign natural history in our population than in Asian populations. Our findings emphasize the need to better understand the natural history of patients with moyamoya as well as the clinical benefit of different treatment modalities. Structured multicenter randomized clinical trials are needed to further assess the best treatment modalities for patients with moyamoya in the United States. PMID- 11493169 TI - Basal ganglia metabolite abnormalities in minor motor disorders associated with human immunodeficiency virus type 1. AB - BACKGROUND: Minor motor disorders (MMDs) associated with human immunodeficiency virus type 1 (HIV-1) predict HIV-1 dementia and death. Little is known about the time course and neuropathologic mechanisms of HIV-1 MMDs. OBJECTIVE: To investigate the relationship between HIV-1 MMDs, as assessed by psychomotor speed, and metabolic alterations in the basal ganglia, as detected by proton magnetic resonance spectroscopy. PATIENTS AND METHODS: A total of 32 HIV-1 seropositive patients (10 with no MMD, 8 with incipient MMD, and 14 with sustained MMD, assessed through electrophysiologic testing of psychomotor speed including contraction times; 29 treated with highly active antiretroviral therapy) and 14 HIV-1-seronegative control subjects were examined for cerebral metabolite abnormalities in the basal ganglia by means of magnetic resonance spectroscopy. RESULTS: The 3 patient groups showed significantly different ratios of myoinositol/creatine (P =.02) in the basal ganglia. Whereas patients with no MMD or incipient MMD showed normal ratios, patients with sustained MMD showed higher values for myoinositol/creatine as a sign of glial proliferation. No differences in N-acetyl compounds, indicative of neuronal loss, were found. CONCLUSION: Whereas metabolic alterations in the basal ganglia were not detected in patients with incipient HIV-1 MMD, patients with sustained HIV-1 MMD did have significantly altered metabolic spectra indicative of glial proliferation. PMID- 11493170 TI - Bilateral sudden deafness as a prodrome of anterior inferior cerebellar artery infarction. AB - BACKGROUND: Acute ischemic stroke in the distribution of the anterior inferior cerebellar artery is known to be associated with hearing loss, facial weakness, ataxia, nystagmus, and hypalgesia. There have been few reports on bilateral deafness and vertebrobasilar occlusive disease. Furthermore, previous reports have not emphasized the inner ear as a localization of bilateral deafness. OBJECTIVE: To describe the presentation of acute ischemic stroke in the distribution of the anterior inferior cerebellar artery as sudden bilateral hearing loss with minimal associated signs. DESIGN AND SETTING: Case report and tertiary care hospital. PATIENT: A 66-year-old man with diabetes mellitus developed sudden bilateral deafness, unilateral tinnitus, and vertigo 7 days before the onset of dysarthria, facial weakness, and ataxia. T2-weighted magnetic resonance imaging scans showed hyperintensities in the right lateral pons and right middle cerebral peduncle and a possible abnormality of the left middle cerebellar peduncle. A magnetic resonance angiogram showed moderately severe stenosis of the distal vertebral artery and middle third of the basilar artery. The patient's right limb coordination and gait improved steadily over several weeks, but there was no improvement in hearing in his right ear. CONCLUSIONS: The relatively isolated onset of deafness as well as the severity and persistence of the hearing loss led us to conclude that the hearing loss in this case was likely due to prominent hypoperfusion of the internal auditory artery, with labyrinthine infarction as the earliest event. Vertebrobasilar occlusive disease should be considered in the differential diagnosis of sudden bilateral deafness. PMID- 11493171 TI - Vascular compression in trigeminal neuralgia shown by magnetic resonance imaging and magnetic resonance angiography image registration. PMID- 11493172 TI - Myelopathic onychodystrophy. PMID- 11493173 TI - The significance of neutralizing antibodies in patients with multiple sclerosis treated with interferon beta. PMID- 11493174 TI - Measurement of antibodies to interferon beta in patients with multiple sclerosis. PMID- 11493175 TI - Commentary on neutralizing antibodies to interferon beta in patients with multiple sclerosis. PMID- 11493176 TI - Brain abscess. PMID- 11493177 TI - The epilepsy of Fyodor Dostoyevsky: insights from Smerdyakov Karamazov's use of a malingered seizure as an alibi. PMID- 11493178 TI - Altered mental state and nonconvulsive status epilepticus in patients with cancer. PMID- 11493180 TI - Linear pontine and trigeminal root lesions and trigeminal neuralgia. PMID- 11493182 TI - Ocular motor apraxia and ataxia-telangiectasia. PMID- 11493198 TI - Localization of the gene for familial laryngeal abductor paralysis to chromosome 6q16. AB - BACKGROUND: Vocal fold paralysis is a common cause of neonatal stridor. Although it is usually classified as idiopathic or iatrogenic in origin, a small subset of patients have a family history of this disorder, indicating a possible genetic cause. OBJECTIVE: To identify the genetic locus of the gene that causes familial laryngeal abductor paralysis. DESIGN: A standard nonorganic protocol was used to extract DNA from whole-blood samples. The DNA samples were quantified by DNA fluorometry, and the concentration of all samples was standardized at 40 ng/microL. A pooled DNA strategy was used to facilitate rapid polymerase chain reaction screening of markers in the Weber v8.0 genome screening set. Polymerase chain reaction screening of individual DNA samples was performed using possible linked markers initially identified as having an allele that appeared with a higher incidence in the affected DNA pools. Statistical analysis of possible linkage was performed using the LINKAGE 5.1 set of linkage analysis computer programs. SUBJECTS: A family in which a form of familial laryngeal abductor paralysis segregates was ascertained. Whole blood samples were drawn from 40 participating individuals within this family after the subjects' fully informed consent was obtained. RESULTS: Initial screening of the pooled DNA specimens revealed a band pattern for D6S1021 on chromosome 6q16, indicating an allele with a higher incidence in the affected vs the nonaffected pool. Two-point analysis of individual allele patterns confirmed linkage to D6S1021 with an lod score of 3.86 (straight theta = 0.0) at a penetrance value of 0.8. Haplotype analysis with flanking markers defined a 5-centiray critical region between D6S283 and AFMA047YG1. CONCLUSION: An autosomal dominant form of familial laryngeal abductor paralysis is linked to a 5-centiray region on chromosome 6q16 surrounding D6S1021. PMID- 11493199 TI - MYO1F as a candidate gene for nonsyndromic deafness, DFNB15. AB - BACKGROUND: Earlier studies have mapped the autosomal recessive nonsyndromic deafness locus, DFNB15, to chromosomes 3q21.3-q25.2 and 19p13.3-13.1, identifying one of these chromosomal regions (or possibly both) as the site of a deafness causing gene. Mutations in unconventional myosins cause deafness in mice and humans. One unconventional myosin, myosin 1F (MYO1F), is expressed in the cochlea and maps to chromosome 19p13.3-13.2. OBJECTIVE: To evaluate MYO1F as a candidate gene for deafness at the DFNB15 locus by determining its genomic structure and screening each exon for deafness-causing mutations to identify possible allele variants of MYO1F segregating in the DFNB15 family. METHODS: We used radiation hybrid mapping to localize MYO1F on chromosome arm 19p. We next determined its genomic structure using multiple long-range polymerase chain reaction experiments. Using these data, we completed mutation screening using single stranded conformational polymorphism analysis and direct sequencing of affected and nonaffected persons in the original DFNB15 family. RESULTS: Radiation hybrid mapping placed MYO1F in the DFNB15 interval, establishing it as a positional candidate gene. Its genomic structure consists of 24 coding exons. No mutations or genomic rearrangements were found in the original DFNB15 family, making it unlikely that MYO1F is the disease-causing gene in this kindred. CONCLUSIONS: Although we did not find MYO1F allele variants in one family with autosomal recessive nonsyndromic hearing loss, the gene remains an excellent candidate for hereditary hearing impairment. Given its wide tissue expression, MYO1F might cause syndromic deafness. PMID- 11493200 TI - Connexin 26 gene mutations in congenitally deaf children: pitfalls for genetic counseling. AB - OBJECTIVE: To evaluate difficulties encountered in genetic counseling in deaf children carrying connexin 26 gene (CX26 or GJB2) mutations. DESIGN: Prospective study. SETTING: Outpatients, tertiary referral center. PATIENTS: Ninety-six unrelated deaf children in whom CX26 mutations had been detected consecutively. Children were recruited to a center for genetic counseling for deaf children, and all had congenital deafness, sporadic or familial. RESULTS: In 63 children, deafness was clearly a DFNB1 form with autosomal recessive inheritance: 47 of the 63 were homozygous for the most frequent mutation, the deletion of G at position 35 (35delG); 16 of 63 carried on both alleles of CX26 frameshift or stop mutations, or missense mutations affecting a critical region of the gene. In 33 of 96 children, genetic counseling was difficult: 21 of 33 had a single mutation detected, 11 of 33 had new missense mutations or mutations whose pathogenicity remains debated in the literature, and 1 of 33 had a genotype with both a recessive mutation (35delG) and a mutation acting as a dominant mutation. CONCLUSIONS: Interpretation of results for the molecular diagnosis of mutations in the connexin 26 gene is difficult in almost one third of cases. Close collaboration between geneticists familiar with deafness and otolaryngologists is essential to provide a high standard of genetic advice. PMID- 11493201 TI - Pediatric otolaryngologists' knowledge and understanding of genetic testing for deafness. AB - OBJECTIVE: To assess the level of a cohort of pediatric otolaryngologists' knowledge and understanding of genetics and genetic testing for deafness and hard of hearing (D/HOH). METHODS: A questionnaire was designed to assess the level of knowledge and understanding of the genetic basis and genetic testing for deafness among a cohort of pediatric otolaryngologists. Three hundred questionnaires were made available to attendees of the 14th (1999) Annual Meeting of the American Society of Pediatric Otolaryngology, Palm Desert, Calif. A series of questions asked to gauge the respondent's level of knowledge of genetics and hearing impairment addressed estimating recurrence risks for deaf and normal-hearing parents and the likelihood of detecting a mutation in connexin 26 in specific clinical scenarios. RESULTS: A total of 28 questionnaires were completed and returned. All respondents reported that they regularly saw patients for D/HOH. Almost half commonly refer these patients for genetic testing and counseling. Seventeen (71%) of 24 otolaryngologists stated they offered genetic testing in all situations, while 6 offered counseling only at parental request or to address recurrence risk issues. One otolaryngologist offered genetic testing if there was a deaf sibling. Twelve (67%) of 18 offered pretest counseling, which was most frequently provided by a genetic counselor. Although 3 (19%) of 16 otolaryngologists provided the counseling themselves, 2 (13%) reported that they and a genetic counselor provided the counseling. While 24 (89%) of the 27 correctly stated that nonsyndromic D/HOH is usually autosomal recessive, recurrence risks were incorrectly estimated in several examples. CONCLUSIONS: While the surveyed pediatric otolaryngologists have a good knowledge of genetics and genetic testing for D/HOH, recurrence risks were often inaccurate. Since D/HOH testing is clinically available, it is imperative that physicians are educated about genetics and genetic testing and are able to communicate this to their patients and their patients' families. PMID- 11493202 TI - Mutation screening for deafness: more than simply another diagnostic test. PMID- 11493203 TI - Different distribution of HLA class II alleles according to response to corticosteroid therapy in sudden sensorineural hearing loss. AB - OBJECTIVE: To investigate the association of HLA class II alleles with the susceptibility to sudden sensorineural hearing loss and with the results of corticosteroid treatment in the Korean population. DESIGN: HLA-DRB1, -DQA1, DQB1, and -DPB1 genotyping by the sequence-specific oligonucleotide probes method in 41 patients with sudden sensorineural hearing loss and in 206 healthy control subjects. Initial hearing levels at the onset of hearing loss and final hearing levels after treatment were evaluated for the association with HLA class II alleles. SETTING: Tertiary care referral center, ambulatory and hospitalized care. SUBJECTS: Forty-one patients (24 men and 17 women; mean age, 49.2 years) were compared with 206 controls. Patients were divided into 2 groups according to their response to corticosteroid therapy (good response vs nonresponse). RESULTS: The frequencies of HLA-DRB1, -DQA1, -DQB1, and -DPB1 alleles were not significantly different between patients and controls (P>.05). When an association between the results of corticosteroid treatment and the frequency of HLA alleles was evaluated, the frequencies of HLA-DRB1*14 (relative risk [RR] = 3.5, P<.02), -DQA1*03 (RR = 4.2, P<.02), and -DQA1*05 (RR = 3.1, P<.03) were significantly increased, but HLA-DQA1*01 (RR = 0.2, P<.004) and -DQB1*06 (RR = 0.2, P<.009) were decreased in the group nonresponsive to corticosteroid therapy, compared with the controls. The distribution of HLA-DQA1*01 (P<.04), -DQB1*06 (P<.02), and -DQA1*03 (P<.003) was significantly different between the responsive and the nonresponsive groups. HLA-DQA1 allelic combination analysis showed that the frequencies of DQA1*03 and *05 had a high RR value in patients with sudden sensorineural hearing loss (RR = 4.1, P<.003) and in patients in the nonresponsive group (RR = 8.9, P<.001), compared with the controls. CONCLUSION: The presence of HLA class II alleles may be a useful genetic marker in forecasting a prognosis in Korean patients with sudden sensorineural hearing loss. PMID- 11493204 TI - Safety of pediatric bedside tracheostomy in the intensive care unit. AB - BACKGROUND: Elective bedside pediatric tracheostomies in the intensive care unit have not been widely reported. Unlike in the adult population, this is not yet considered a safe or routine procedure in the pediatric population. We performed a preliminary study suggesting that bedside pediatric tracheostomies can be done safely and at reduced cost. DESIGN: Retrospective medical chart review. SETTING: Tertiary care referral center at a single university hospital. PATIENTS: Fifty seven patients, ranging in age from 15 days to 8 years. Thirty operating room tracheostomies and 27 bedside tracheostomies were performed during a 6-year period. The mean age of the patients was 20.5 months, with no significant age difference between the 2 groups. The top 3 diagnoses necessitating tracheostomy were laryngotracheal disorders (18 patients [32%]), bronchopulmonary dysplasia (9 [16%]), and neurologic disorders (6 [11%]). INTERVENTIONS: Tracheostomy. MAIN OUTCOME MEASURES: The initial 48-hour postoperative period was examined to compare complication rates between groups. RESULTS: Overall, the 2 groups had similar complication rates (chi(2) = 0.12; P =.73). The operating room group had 3 complications (10%) related to bleeding, infection, and pneumothorax. The bedside group had 2 complications (7%), both involving pneumothorax. Each operating room tracheostomy incurred charges totaling $1693 vs $235 for each bedside tracheostomy. CONCLUSIONS: Historically, pediatric tracheostomy has been viewed as a technically demanding procedure with a high complication rate, thus encouraging routine operating room use. We found that pediatric tracheostomy performed in the intensive care unit, with attention to prudent patient selection and adherence to consistent, sound techniques, was as safe as operating room tracheostomy. PMID- 11493205 TI - Effects of the Clarion Electrode Positioning System on auditory thresholds and comfortable loudness levels in pediatric patients with cochlear implants. AB - OBJECTIVE: To evaluate the effects of using the Electrode Positioning System on psychophysical auditory thresholds, most comfortable loudness levels, and electric auditory brainstem response (EABR) thresholds in children with the Clarion version 1.2 cochlear implant. DESIGN: Retrospective analysis. SETTING: Academic tertiary care center. PATIENTS AND METHODS: Clinical records of a series of 25 children who received the Clarion version 1.2 cochlear implant at the University of Minnesota, Minneapolis, between January 1997 and August 1999 were examined. Measures evaluated were psychophysical thresholds (T-levels) and most comfortable loudness levels (M-levels) obtained at the 3-month posthookup audiologic evaluation and EABR thresholds obtained during implant surgery. Relevant threshold measures were available for 24 patients, 11 of whom had received the Clarion spiral electrode and electrode positioner (EP group) and 13 of whom had received the spiral electrode without positioner (non-EP group). The 3 measures (T-levels, M-levels, and EABR thresholds) were compared across groups. In addition, EABR thresholds were compared with T-levels and M-levels within groups. RESULTS: Mean T-levels and M-levels were significantly lower for the EP group than for the non-EP group, and interpatient variability for these measures was considerably smaller in the EP group. Electric auditory brainstem response thresholds were not significantly different for EP vs non-EP patients; however, EABR data were available for only a few non-EP patients. CONCLUSIONS: Use of the electrode positioner results in lower T-levels and M-levels in children with the Clarion version 1.2 cochlear implant, consistent with results of previous studies in adults, and reduces across-patient variability for these measures. It is unclear from the present data whether use of the electrode positioner systematically reduces intraoperative EABR thresholds. PMID- 11493206 TI - Measurement of comfort levels by means of electrical stapedial reflex in children. AB - BACKGROUND: Patient success and satisfaction with a cochlear implant largely depend on the adequacy of the speech-processing program. The program is generated by means of behaviorally determined threshold and comfort levels for each electrode. As the minimum age for implantation continues to decrease, behavioral methods of measuring comfort levels have become more problematic, and so the need for objective ways to program speech processors has become more important. OBJECTIVES: To evaluate the use of electrically evoked stapedial reflexes (ESRs) to measure comfort levels for children and compare these results with behavioral measurements, and to report the results of a questionnaire assessing the acceptability and general performance of program before and after adjustment of comfort levels measured with ESRs. DESIGN AND SETTING: Before-after trial in the cochlear implant unit of a tertiary hospital. PATIENTS AND METHODS: Programming with the ESR technique was successfully completed in 20 of a consecutive sample of 26 children undergoing programming of their cochlear implants. OUTCOME MEASURES: Programming units as measured by the 2 programming techniques and numerical score of questionnaire. RESULTS: Comfort levels with the ESR method were found to be consistently lower than those obtained with behavioral techniques. Children using programs set with ESRs wore their implants longer and had fewer episodes of discomfort to environmental sounds. CONCLUSION: Comfort level estimation by means of ESRs is reliable and objective and hence a valuable programming tool in the pediatric population. PMID- 11493207 TI - Younger patients have a higher rate of recovery of taste function after middle ear surgery. AB - BACKGROUND: Although the chorda tympani nerve (CTN) is frequently damaged during tympanoplasty, little attention has been given to the patients' symptoms and taste function. OBJECTIVE: To investigate patients' symptoms and the functional recovery of taste after surgery using electrogustometry (EGM). DESIGN: Prospective study. PATIENTS: Analysis of 163 ears of 156 patients who underwent middle ear surgery from April 1997 through December 1999. There were 18 ears with noninflammatory diseases, 80 with chronic otitis media, and 65 with cholesteatoma. The patients' taste functions were examined 2 days before surgery and 2 weeks and 6 months after surgery. MAIN OUTCOME MEASURES: The taste disturbance before and after middle ear surgery and the relationship between age and the recovery rate of CTN function. RESULTS: Two weeks after surgery, the mean EGM threshold was elevated in all groups regardless of preservation or section of the CTN. Numbness in the tongue and taste disturbance were more frequently found in patients with preservation of CTN than in those with section of the CTN (P =.008 and P =.001, respectively). In patients with preservation of the CTN, 6 months after surgery, the recovery rate of EGM threshold was 83% in those aged 20 years or younger (P =.008 compared with the 2 older groups), 53% in the those aged 21 to 40 years, and 44% in those aged 41 to 60 years. CONCLUSION: Age is an important factor for recovery of taste function after middle ear surgery, which is useful information when explaining complications to patients. PMID- 11493208 TI - Sentinel node localization in oral cavity and oropharynx squamous cell cancer. AB - OBJECTIVE: To evaluate the feasibility and predictive ability of the sentinel node localization technique for patients with squamous cell carcinoma of the oral cavity or oropharynx and clinically negative necks. DESIGN: Prospective, efficacy study comparing the histopathologic status of the sentinel node with that of the remaining neck dissection specimen. SETTING: Tertiary referral center. PATIENTS: Patients with T1 or T2 disease and clinically negative necks were eligible for the study. Nine previously untreated patients with oral cavity or oropharyngeal squamous cell carcinoma were enrolled in the study. INTERVENTIONS: Unfiltered technetium Tc 99m sulfur colloid injections of the primary tumor and lymphoscintigraphy were performed on the day before surgery. Intraoperatively, the sentinel node(s) was localized with a gamma probe and removed after tumor resection and before neck dissection. MAIN OUTCOME MEASURES: The primary outcome was the negative predictive value of the histopathologic status of the sentinel node for predicting cervical metastases. RESULTS: Sentinel nodes were identified in 9 previously untreated patients. In 5 patients, there were no positive nodes. In 4 patients, the sentinel nodes were the only histopathologically positive nodes. In previously untreated patients, the sentinel node technique had a negative predictive value of 100% for cervical metastasis. CONCLUSIONS: Our preliminary investigation shows that sentinel node localization is technically feasible in head and neck surgery and is predictive of cervical metastasis. The sentinel node technique has the potential to decrease the number of neck dissections performed in clinically negative necks, thus reducing the associated morbidity for patients in this group. PMID- 11493209 TI - Prediction of aspiration in patients with newly diagnosed untreated advanced head and neck cancer. AB - OBJECTIVES: To determine the prevalence of aspiration in patients with newly diagnosed nontreated advanced head and neck cancer and to determine the ability of the clinical examination to predict aspiration in this patient population. DESIGN: A descriptive prevalence study of the presence of aspiration in patients with advanced (stages III and IV) head and neck cancer and a prospective correlation study between variables of the clinical evaluation with the criterion standard videofluoroscopy. SETTING: Regional veterans affairs medical center. PATIENTS: A consecutive sample of 27 patients without previous treatment or tracheostomy. INTERVENTIONS: All patients underwent evaluation by an otolaryngologist and speech pathologist prior to videofluoroscopy. MAIN OUTCOME MEASURES: Variables in the dysphagia evaluation (consisting of a directed medical history, physical examination, and food challenge), the self-rated 45-item questionnaire, and the clinicians "educated prediction" of aspiration were analyzed with reference to aspiration on videofluoroscopy using univariant analysis. RESULTS: The prevalence of aspiration determined by videofluoroscopy was 41% (11 of 27 patients). Two (6%) of 32 items in the medical history, 1 (4%) of 25 items in the physical examination, and 8 (50%) of 16 items in the food challenge were found to be statistically significantly correlated (P<.05) with aspiration on videofluoroscopy by univariate analysis. Nine (25%) of 45 items in the self-rated questionnaire were correlated with aspiration on videofluoroscopy. The clinicians' educated prediction of aspiration was not correlated with aspiration determined by videofluoroscopy. CONCLUSIONS: Clinical evaluation alone is inadequate in predicting patients who aspirated, determined objectively by videofluoroscopic swallowing study. Further study is needed to develop an efficient dysphagia evaluation to identify patients at risk for aspiration. PMID- 11493210 TI - Efficacy of gastric aspiration in reducing posttonsillectomy vomiting. AB - OBJECTIVE: To determine the effectiveness of postoperative gastric decompression in reducing the incidence and complications associated with vomiting following tonsillectomy. DESIGN: A prospective, randomized controlled study. SETTING: Private office and clinic of a university teaching hospital and research center. PATIENTS: Eighty pediatric patients ranging in age from 22 months to 11 years, American Society of Anesthesiologists class I or II, undergoing tonsillectomy with or without adenoidectomy were enrolled in the study. Six were excluded from the final analysis, 5 because of failure of the parents to complete and return the data forms and 1 because of postoperative bleeding. Of the 74 patients included in the study, 35 were in the control group and 39 were in the study group. INTERVENTIONS: The 39 patients in the study group underwent postoperative aspiration of gastric contents with an orogastric tube placed under direct visualization while the patient was still under general anesthesia. The 35 patients in the control group did not undergo gastric aspiration following surgery. MAIN OUTCOME MEASURES: The incidence of vomiting, the number of episodes of vomiting before and after hospital discharge, the total volume of emesis, the postoperative length of stay, the need for rescue antiemetic prophylaxis, and the number of readmissions to the hospital for persistent vomiting were noted. RESULTS: No statistically significant difference (P<.05) was noted between the control group and the study group for the percentage of patients experiencing vomiting (74% vs 85%), the mean number of episodes of vomiting before (2.6 vs 2.8) and after (0.8 vs 0.7) hospital discharge, the mean volume of emesis (157 mL vs 222 mL), the postoperative length of stay (394 minutes vs 334 minutes), the percentage of patients requiring rescue antiemetics (34% [12 patients] vs 33% [13 patients]), and the percentage of unplanned admissions because of vomiting (9% [3 patients] vs 15% [6 patients]). CONCLUSION: Our results indicate that gastric aspiration does not decrease the incidence of vomiting following tonsillectomy. PMID- 11493211 TI - C57Bl/6 and BALB/c mice have similar neutrophil response to acute Streptococcus pneumoniae sinus infections. AB - BACKGROUND: Previous investigations have shown that mice with a tendency toward a T(H)1 or T(H)2 lymphocyte response manifest different reactions to inoculation with the parasite Leishmania major. BALB/c mice (with a tendency for a T(H)2 response) showed evidence of systemic infection, whereas C57Bl/6 mice (with a tendency for a T(H)1 response) showed only a local reaction. OBJECTIVE: To investigate whether BALB/c and C57Bl/6 mice respond differently to acute bacterial infection of the sinuses. METHODS: We inoculated the nasal cavities of C57Bl/6 and BALB/c mice with Streptococcus pneumoniae (type ATCC59), or with broth as a control. The mice were humanely killed 2, 5, 10, and 14 days after inoculation. Their heads were fixed, decalcified, and embedded in paraffin blocks. Sections were stained with hematoxylin and eosin, and the degree of inflammation was quantified by the number of neutrophils per square millimeter of the sinus mucosa and the percentage of the sinus cavity occupied by neutrophil clusters. RESULTS: Both groups of mice showed evidence of inflammation that was significantly greater than controls (P =.01), with no difference between groups. There was a correlation between the number of neutrophils per square millimeter in the sinus mucosa and the percentage of neutrophil clusters (C57Bl/6 mice, r = 0.37, P<.001; BALB/c mice, r = 0.20, P<.001). In the infected mice, the number of infiltrating neutrophils was significantly greater (P<.001) in anatomically lower (dependent) areas of the sinuses compared with the upper areas. CONCLUSION: Unlike leishmaniasis, acute bacterial sinusitis is not affected by the tendency of the host to favor either a T(H)1 or T(H)2 response. PMID- 11493212 TI - Microvascular transplantation and replantation of the rabbit submandibular gland. AB - BACKGROUND: Xerostomia is a devastating complication of radiation therapy. Previous research has demonstrated that submandibular glands may be removed from the neck and transplanted using microvascular techniques, with good gland survival. However, microvascular transplantation and replantation has never been attempted on a composite tissue such as a salivary gland. OBJECTIVE: To evaluate the ability of a rabbit submandibular gland to undergo 2 successive microvascular transplantations. SUBJECTS AND DESIGN: Study rabbits underwent a midline neck incision with dissection of a submandibular gland to its arterial and venous pedicle. Microvascular techniques were then used to transplant the gland to the femoral system of the right groin. The incisions were reopened later under surgical conditions. The transferred gland was examined for survival and patency of its artery and vein. Healthy glands were dissected and transferred to a suitable artery and vein within the neck, where they were again reanastamosed using microvascular surgical techniques. After additional time, the gland was again examined for survival and pedicle patency, then removed and evaluated for histopathological evidence of survival. RESULTS: Surgical technique evolved during the course of this work to avoid encountered pitfalls. After refining the technique, we have determined that the rabbit submandibular gland is able to withstand successive microvascular transplantation and replantation with good likelihood of long-term survival, according to histopathological criteria. CONCLUSIONS: The rabbit submandibular gland is able to undergo microvascular transplantation and replantation with evidence of long-term survivability and preserved function. The body's natural response to surgery and tissue transplantation makes replantation a technical challenge; however, methods delineated herein alleviate many of the potential pitfalls. Extending these results to humans, patients who are to undergo radiation therapy could have a disease-free gland removed from the neck, transferred outside of the field of radiation, and then returned to the neck at the completion of radiation therapy. This may enable them to maintain salivary gland function and maintain oral cavity function and comfort. PMID- 11493213 TI - Heterotopic neuroglial tissue causing airway obstruction in the newborn. AB - BACKGROUND: Heterotopic neuroglial (brain) tissue is a rare cause of airway obstruction in newborns. Fewer than 30 cases have been reported in the English literature. Brain heterotopias can mimic more common congenital anomalies of the head and neck. OBJECTIVE: To review our experience in the diagnosis and treatment of children with heterotopic pharyngeal neuroglial tissue. DESIGN: Case series. SETTING: Tertiary care children's hospital. PATIENTS: Four newborns with airway obstruction caused by heterotopic neuroglial tissue. RESULTS: All patients were infants (3 full-term girls and a 32 weeks' gestation boy) who had airway obstruction in the newborn period. All patients underwent preoperative computed tomography and magnetic resonance imaging, which revealed a heterogeneous mass involving the pharynx, neck, and parapharyngeal space. Bony deformities of the skull base and mandible were present in all patients, although intracranial connection was absent. Multiple surgical procedures were performed in all 4 patients. Tracheotomy was performed in 2 patients, gastrostomy tube placement was required in 3, and a nasopharyngeal tube was used in 1. Combined cervicofacial and transoral approaches were used for resection, preserving vital structures. Histopathologic evaluation revealed mature glial tissue and choroid plexus-like structures. CONCLUSIONS: Heterotopic neuroglial tissue must be considered in the differential diagnosis of airway obstruction in the newborn. Management is surgical resection, with attention to vital structures and function-analogous to surgery for lymphangioma. Multiple surgical procedures might be necessary in the treatment of these patients. PMID- 11493214 TI - Pathology quiz case: plasmacytoid variety of myoepithelioma of minor salivary gland. PMID- 11493215 TI - Radiology quiz case: eosinophilic granuloma. PMID- 11493216 TI - Perioperative intravenous steroid treatment and tonsillectomy. PMID- 11493217 TI - Tonsillectomy and postoperative vomiting: Do steroids really work? PMID- 11493218 TI - Do intravenous steroids play a role for tonsillectomy patients? PMID- 11493219 TI - Vascular parkinsonism: a distinct, heterogeneous clinical entity. AB - OBJECTIVES: The aim of this study was to define the symptoms and signs of suspected vascular parkinsonism (VP) which is still a debatable concept. MATERIAL AND METHODS: Patients with parkinsonism were grouped into patients with suspected VP and Parkinson's disease (PD) after other causes for secondary parkinsonism, and parkinsonism-plus syndromes were excluded. The clinical features of 16 patients with suspected VP to those of 50 diagnosed with PD were compared. All patients were assessed using unified Parkinson's disease rating scale (UPDRS) and all had cerebral MRIs. RESULTS: Patients with VP had significantly older onset age and shorter duration of disease with gait disorder as the most frequent initial symptom. All PD patients had satisfactory response to levodopa treatment, whereas only 38% VP patients had satisfactory response to levodopa treatment. Vascular risk factors were more common in VP (81%) than PD (32%). Postural instability, freezing, gait disturbance, pyramidal signs, and postural tremor were significantly more prevalent in patients with VP than in PD. In VP patients these features were more prominent in the lower limbs. Twenty-five percent had acute onset VP. All patients with VP had ischemic lesions, mainly in subcortical white matter, to a lesser extent basal ganglia and brainstem, in their cerebral MRIs, while 70% of PD patients had normal MRIs. CONCLUSION: The differences in the clinical features support the concept that VP is a distinct clinical entity with heterogeneous clinical, MRI, and possibly pathophysiological features. PMID- 11493220 TI - Evidences of vascular origin of cochleovestibular dysfunction. AB - OBJECTIVES: When the vascular disorder of the cochleovestibular system is mentioned, the diagnosis is based on exclusion of other diseases. Since arteries of the cochleovestibular system cannot be directly visualized, physicians must deduce from the vascular risk factors and the vascular lesion of other territories to the vascular cochleovestibular disease. MATERIALS AND METHODS: Authors analyzed the data of 19 patients with vertigo. Detailed blood tests, complete neurootological and audiological examination including ABR, carotid and vertebral artery Doppler sonography, MRI and MRA was performed. RESULTS: Cochleovestibular examination and ABR showed abnormalities in 73.7%, either carotid and vertebral artery Doppler or MRI showed abnormalities in 57.9%. MRA was abnormal in 47.4%. In most of the patients multiple risk factors of cerebrovascular disorder could be found. CONCLUSIONS: The cochleovestibular system disorders can be considered to be of vascular origin if the examinations exclude other diseases, if the patients have vascular risk factors and if other territories of brain accessible for imaging methods show vascular disorders. PMID- 11493221 TI - Paraneoplastic neurological syndromes - patients' cohort profile in the Czech Republic. AB - Reported paraneoplastic neurological syndromes (PNS) are rare disabling neurological diseases with supposed autoimmune pathogenesis. The aims of this study were to evaluate frequency, clinical course and therapeutic response in the cohort of PNS positive patients (n=10) in the Czech Republic for the first time. Second, we determined the presence and distribution of oligoclonal IgG bands (OB IgG) in PNS and compared the clinical and laboratory features of OB IgG positive and negative patients. A total of 2355 suspicious serum and/or CSF samples were screened by immunofluorescence and immunohistochemistry with definite confirmation by Western blot. OB IgG were detected by isoelectric focusing and immunoenzymatic staining and clinical status was scored according to modified Rankin scale (RS). Four patients had anti-Yo antibody, ovarian cancer and the score in range (2-5) on RS. Five patients had anti-Hu antibody, small cell lung cancer (SCLC), prostate cancer and the score between 1-4 grade on RS. One patient with SCLC and anti-Ri antibody had grade 2. Five of 10 patients with PNS had positive OB IgG and average value 4.2 on RS comparing with negative OB IgG patients with average value 2.6. Finally, we add well-defined cohort of PNS patients to emerging European profile of PNS and conclude that the presence of OB IgG in PNS seems to reflect enhanced immune response with more severe neurological damage and clinical course. PMID- 11493222 TI - Double filtration plasmapheresis in the treatment of myasthenic crisis--analysis of prognostic factors and efficacy. AB - OBJECTIVES: To examine the prognostic factors and outcome of myasthenia gravis (MG) patients in crisis with double filtration plasmapheresis (DFP) treatment. MATERIAL AND METHODS: A total of 15 patients experienced 20 episodes of crisis during the study period. Plasmapheresis was carried out using a double filtration METHOD: Demographic information, clinical features of crisis, and associated complications were analyzed. RESULTS: The median duration of crisis was 9 days. Chest infection was the most common precipitant of crisis. Twelve out of the 20 episodes (60%) responded well to DFP and mechanical ventilation was discontinued after the third session of DFP in 8 of them. Three significant predictors for prolonged crisis were shorter intervals between the onset of MG and the first crisis (P=0.04), higher serum bicarbonate levels at baseline (P=0.03) and the thymic pathology of thymoma (P=0.03). CONCLUSION: DFP can ameliorate the profound weakness in crisis and seems to be a rational therapy for patients with myasthenic crisis. PMID- 11493223 TI - Frequency and significance of anti-Ro (SS-A) antibodies in multiple sclerosis patients. AB - OBJECTIVE: To determine the frequency and significance of antinuclear (ANA), anticardiolipin (ACA) and anti-Ro (SS-A) antibodies in multiple sclerosis (MS) patients. METHODS: ANA (indirect immunofluorescence), ACA and anti-Ro (SS-A) antibodies (ELISA) were tested in sera of 42 patients with Poser defined MS and 50 healthy individuals. RESULTS: High levels of anti-Ro (SS-A) antibodies were found in 3 patients (7%) (vs 0 in the control group). Two of them had normal salivary gland biopsy. Clinical MS form was chronic-progressive in 2 cases and relapsing-remitting in the third one. Ten patients (23%) had low levels of ANA (vs 4%), none of them positive for anti-Ro (SS-A) antibodies. Only 1 patient (2%) with RR clinical form had ACA (vs 0). No clinical or neuroradiological differences with conventional MS patients were observed. CONCLUSIONS: ANA, ACA and anti-Ro (SS-A) antibodies in MS patients indicate an underlying autoimmune disease but our series suggests that they are an epiphenomenon of a more diffuse immunological dysfunction. PMID- 11493224 TI - Differential release of beta-chemokines in serum and CSF of patients with relapsing-remitting multiple sclerosis. AB - OBJECTIVE: beta-chemokines were recently demonstrated in active MS-lesions. We tested whether MCP-1 and RANTES can also be detected in CSF and serum of patients with MS and whether release is associated with inflammatory disease activity. MATERIALS AND METHODS: CSF and serum from 34 patients with newly diagnosed relapsing-remitting MS (RR-MS), 17 patients with viral meningitis (VM) and 19 patients with non-inflammatory neurological diseases (NIND) were investigated by ELISA. RR-MS patients underwent lumbar puncture and Gd-enhanced MRI examinations within 2 days. RESULTS: MCP-1 was strong intrathecally released in all patients. Compared to NIND CSF-levels were increased in VM (P<0.001) and were decreased in RR-MS (P<0.05). RANTES was only detected in serum in all patients. Levels were higher in VM and RR-MS compared to NIND (P<0.05). A total of 14/34 RR-MS patients exhibited active Gd-enhancing lesions on MRI. They had lower MCP-1 levels in CSF (P<0.001) and serum (P<0.05) and higher serum levels of RANTES (P<0.05) as compared to patients without active lesions. CONCLUSIONS: MCP-1 and RANTES are differentially released during acute attacks of RR-MS, which might reflect different immunregulatory roles of these beta-chemokines in RR-MS. PMID- 11493225 TI - Cardiovascular effects of domperidone in patients with Parkinson's disease treated with apomorphine. AB - OBJECTIVES: The cardiovascular effects of the peripheral dopamine D2-receptor antagonist domperidone were studied in patients with idiopathic Parkinson's disease treated with continuous subcutaneous infusion of apomorphine. MATERIAL AND METHODS: Blood pressure and heart rate of 10 patients were monitored for 24 h before and after treatment with domperidone using an automatic device for blood pressure recording. RESULTS: Blood pressure and heart rate increased significantly during 24 h of domperidone treatment. When day and night values were analysed separately, a significant difference of blood pressure between the groups was found only during the day whereas heart rate was significantly increased during both day and night. Domperidone shifted blood pressure and heart rate upwards without affecting the drop of blood pressure in the erect position. CONCLUSION: Domperidone increases blood pressure and heart rate without inducing nocturnal hypertension in apomorphine treated patients with idiopathic Parkinson's disease. Increased heart rate by domperidone is a new finding but the clinical significance is unclear and requires further investigation. PMID- 11493226 TI - Recombination breakpoints in the Charcot-Marie-Tooth 1A repeat sequence in Norwegian families. AB - OBJECTIVE: To investigate the recombination breakpoint in a 3.2 kb junction fragment of the 24 kb CMT1A repeat sequences (CMT1A-REPs) on chromosome 17p11.2 12. MATERIALS AND METHODS: Thirty-eight Norwegian CMT1 patients and 15 asymptomatic family members of 15 separate families including 10 normal controls were investigated using repeat (REP)-PCR. RESULTS: Twenty-six (68.4%) of the CMT1 patients from 9 (60%) families were positive for the CMT1A duplication which was not found in any of the controls. In 89.9% of the REP-PCR positive families the recombination breakpoint was mapped to a 1.7 kb "hot-spot" region, and in 11.1% of the families to a 1.5 kb region telomeric to the 1.7 kb region. CONCLUSION: The frequency and regions for CMT1A-REPs crossover events in Norwegian CMT1A cases are similar to what is found in other populations. REP-PCR is not, however, as sensitive as other diagnostic methods to detect the CMT1A duplication. PMID- 11493227 TI - HTLV-1 associated polymyositis in Jamaica. AB - The clinical, laboratory and epidemiological characteristics of 38 adult Jamaican patients with polymyositis were evaluated. Twenty-four patients (63%) were human T-lymphotropic virus 1 (HTLV-1) seropositive and 14 patients (37%) were HTLV-1 seronegative. Polymyositis runs a more protracted course in seropositive patients who had more frequent hospital admissions and a significantly longer duration of symptoms prior to presentation. Joint swelling, chest pain and dyspnoea were more frequent complaints among the seronegative patients. There was no significant difference between the two serological groups in muscle enzyme levels, antinuclear antibody positivity or frequency of Jo-1 antibodies. HTLV-1 infection may define a subgroup of polymyositis patients with a more insidious presentation and poorer response to corticosteroid therapy. PMID- 11493228 TI - Brain dysfunction explored by long latency reflex: a study of adrenomyeloneuropathy. AB - OBJECTIVES: We can assess brain function by measuring the cortical relay time (CRT) of long latency reflex (LLR) of hand muscle. We would study if measurement of CRT of LLR can explore the brain involvement of adrenomyeloneuropathy (AMN). METHODS: Two AMN patients were included in the study. Both of them had spastic gait and mild sensory deficits but normal mental function. The LLRs were provoked at the first dorsal interosseous muscle by electrical stimulation of the middle finger. We measured the latency of LLR and its CRT. RESULTS: Delayed LLR and prolonged CRT were noted in AMN patients, even though the magnetic resonance imaging of brain did not show any significant abnormalities. CONCLUSIONS: Measuring CRT of LLR reveals brain involvement of AMN patients, and it is an adjunct in the assessment of brain function though without specific anatomic diagnosis. PMID- 11493230 TI - Blepharoclonus and Arnold-Chiari malformation. AB - OBJECTIVE: Blepharoclonus (BLC) denotes a large amplitude, involuntary tremors of the orbicularis oculi muscles, observed during gentle closure of the eyelids. BLC may follow major head trauma. Four patients with Arnold-Chiari malformation (ACM) and BLC are described. MATERIALS AND METHODS: The first patient had facial numbness for 5 months; the remaining patients had headaches following minor head or cervical spinal injuries. Brain magnetic resonance imaging (MRI), electroencephalogram (EEG) blink reflexes, mental and facial nerve responses and facial electromyogram (EMG) were performed. RESULTS: All patients exhibited ACM on brain MRI. The first patient had coincidental dural venous malformation, empty sella turcica and familial digital dysplasia. She exhibited oculopterygoid synkinesis. The last 3 patients had posttraumatic headache; the second and third patients had limited features of Ehlers-Danlos syndrome (EDS). The second patient had cervical spinal fusion and the fourth a cervical syrinx. All the patients had BLC on gentle eyelid closure. CONCLUSION: BLC is an underdiagnosed neuro ophthalmological sign of ACM. PMID- 11493229 TI - Gabapentin in the treatment of hemifacial spasm. AB - OBJECTIVES: To evaluate the efficacy of gabapentin in the treatment of hemifacial spasm. MATERIAL AND METHODS: Twenty-three patients with hemifacial spasm not suitable for surgery or therapy with botulinum toxin were treated with gabapentin. The main efficacy parameter was the percentage of spasm reduction. RESULTS: A clinically significant reduction of spasms was obtained by 16 patients. CONCLUSION: Gabapentin was effective and safe in reducing hemifacial spasm in 16 out 23 (69.6%) patients. PMID- 11493232 TI - Resuscitating audit. PMID- 11493231 TI - Alpha-like activity in terminal stage of Creutzfeldt-Jakob disease. AB - We report a case of Creutzfeldt-Jakob disease showing various changes in electroencephalogram (EEG) throughout the course of the disease. The patient's EEG patterns showed periodic synchronous discharge in the intermediate stage of the disease, delta activity in the advanced stage and alpha-like activity in the terminal stage. The mechanism generating alpha-like activity may resemble, at least in part, that of an alpha coma. PMID- 11493233 TI - Risk factors for postoperative anxiety in adults. AB - We identified risk factors for postoperative anxiety and quantified their effect on 712 adults between 18 and 60 years of age (ASA I-III physical status) undergoing elective surgery under general anaesthesia, neural blockade or both. The measuring instruments were a structured questionnaire, a pain visual analogue scale, the McGill Pain Questionnaire, the State-Trait Anxiety Inventory, the Montgomery-Asberg Depression Rating Scale, a Self-Reporting Questionnaire-20, and a Self-Perception of Future Questionnaire. Multivariate conditional regression modelling taking into account the hierarchical relationship between risk factors revealed that postoperative anxiety was associated with ASA status III (OR = 1.48), history of smoking (1.62), moderate to intense postoperative pain (OR = 2.62) and high pain rating index (OR = 2.35), minor psychiatric disorders (OR = 1.87), pre-operative state-anxiety (OR = 2.65), and negative future perception (OR = 2.20). Neural block anaesthesia (OR = 0.72), systemic multimodal analgesia (OR = 0.62) and neuroaxial opioids with or without local anaesthesia (OR = 0.63) were found to be protective factors against postoperative anxiety. PMID- 11493234 TI - Matrix metalloproteinase-9 concentrations in critically ill patients. AB - Matrix metalloproteinase-9 is involved in the processing of cytokines during sepsis. We studied 10 critically ill patients within 12 h of fulfilling the American Consensus Conference criteria for severe sepsis and compared the results with 12 critically ill non-septic control patients and eight healthy subjects. Total matrix metalloproteinase-9 concentrations were measured on days 1, 2 and 3. The median admission Acute Physiological and Chronic Health Evaluation II score was 19.5 (range 13-27) in the septic patients and 20.5 (range 7-28) in the non septic patients. Four patients from each group died within 28 days. Matrix metalloproteinase-9 concentrations were elevated significantly in both groups of patients compared with healthy subjects (p = 0.0004) but there was no difference between patients with and without sepsis. Matrix metalloproteinase-9 levels did not change with time, and there was no difference between survivors and those who died. We conclude that matrix metalloproteinase-9 represents a non-specific marker of systemic inflammation. PMID- 11493235 TI - Hypertension as a risk factor for cerebral injury during cardiopulmonary bypass. Protein S100B and transcranial Doppler findings. AB - We studied 22 patients aged 53-78 years scheduled for cardiac surgery under cardiopulmonary bypass. Blood pressure, cardiac output, transcranial Doppler blood flow velocity, arterial blood gases, body temperature and protein S100B, as a marker for cerebral integrity, were evaluated in normotensive and hypertensive patients. Pre-operative mean (SD) arterial blood pressure was 93 (11) mmHg in the normotensive group compared with 116 (15) mmHg in the hypertensive group. We found an increase in protein S100B levels in both groups. Serum protein S100B concentrations in the hypertensive group were significantly higher than in the normotensive group (p < 0.001). The highest mean (SD) values were 2.04 (0.65) micromol x l(-1) in the normotensive group and 7.02 (4.55) micromol x l(-1) in the hypertensive group. These results suggest that cardiopulmonary bypass is associated with a significantly higher rate of cerebral injury in hypertensive patients than in normotensive patients. This may be due to altered autoregulation and insufficient cerebral perfusion. Modifications of cardiopulmonary bypass management for hypertensive patients might be made to decrease the risk of cerebral injury. PMID- 11493236 TI - A mixture of alfentanil and morphine for rapid postoperative loading with opioid: theoretical basis and initial clinical investigation. AB - Pharmacokinetic modelling of estimated central nervous system concentrations was used to devise the optimal mixture of morphine and alfentanil for the treatment of postoperative pain. Modelling revealed that an intravenous opioid pain protocol using an alfentanil-morphine mixture in the proportions 0.75 : 10 mg would provide a profile of analgesia of rapid onset, yet slow offset. The regimen was evaluated in 58 patients in the recovery ward who were randomly allocated to receive analgesia using pain protocols with either morphine or the mixture. Groups were well matched for age, weight and initial pain scores. The mean (SD) time to patient comfort was 27.6 (20.2) min for the mixture and 41.2 (18.6) min for morphine (p = 0.01). Multiple regression analysis revealed that that initial pain score (p = 0.009) and drug group (p = 0.02), but not age, weight or gender were independent predictors of the time to comfort. Drug group was not a significant predictor of adverse effects. PMID- 11493237 TI - Risk perception and communication: recent developments and implications for anaesthesia. AB - This review begins by outlining the history of probability theory, exposing cultural differences between scientists and lay people in the way risks are viewed. The basic principles of the science of risk perception are described, and the various methods of communicating risk in health care, both verbal and numerical, are then discussed critically. These concepts are then applied to the practice of anaesthesia. Risk perception may affect anaesthetists' choice of career and may be involved in the genesis and evolution of critical incidents; we also discuss possibilities for training in risk perception issues. The place of risk communication in informed consent and its ethical implications are discussed. PMID- 11493238 TI - A comparison of two techniques for manual ventilation of the lungs by non anaesthetists: the bag-valve-facemask and the cuffed oropharyngeal airway (COPA) apparatus. AB - In order to evaluate the possible role of the cuffed oropharyngeal airway during cardiopulmonary resuscitation, we compared its use by non-anaesthetists with the bag-valve-facemask technique of providing ventilation. A group of anaesthetic nurses and operating department practitioners were asked to manually ventilate the lungs of 40 patients undergoing elective surgery following the induction of general anaesthesia with neuromuscular blockade. Ventilation was first attempted using the bag-valve-facemask technique and then using the appropriate size cuffed oropharyngeal airway and self-inflating bag. Ventilation was clinically adequate in 32/40 (80%) patients using the bag-valve-facemask and in 38/40 (95%) patients using the cuffed oropharyngeal airway. Measured expired tidal volumes were greater using the cuffed oropharyngeal airway than with bag-valve-facemask ventilation in two-thirds of patients, despite a higher incidence of audible air leak. Successful ventilation was achieved using the cuffed oropharyngeal airway in seven of the eight patients in whom bag-valve-facemask ventilation was inadequate. The cuffed oropharyngeal airway may offer an effective method of providing ventilation during cardiopulmonary resuscitation by non-anaesthetic hospital staff, particularly when attempted ventilation using a bag-valve facemask technique is proving ineffective. PMID- 11493239 TI - Point-of-care testing apparatus. Measurement of coagulation. AB - Point-of-care testing of coagulation parameters provides a more rapid assessment of test results compared with laboratory testing. A new coagulation monitor (GEM PCL, Instrumentation Laboratory, Kirchheim, Germany) was evaluated. Point-of-care data for activated partial thromboplastin time and prothrombin time (expressed as the international normalised ratio) and turn-around-time were compared. Coagulation parameters were compared in the blood of 57 patients with and without heparin therapy. The point-of-care and laboratory test results showed a bias (SD) of -0.26 (4.55) s for activated partial thromboplastin time and -0.011 (0.150) s for prothrombin time. The average turn-around-time was 3 min for point-of-care testing vs. 52 min for laboratory testing. We conclude that the reliability of point-of-care testing is sufficient for clinical use. PMID- 11493240 TI - Undiagnosed adult diastematomyelia associated with neurological symptoms following spinal anaesthesia. AB - Spinal anaesthesia is a safe, routinely performed procedure. However, in patients with lumbar cutaneous abnormities or deformities of the lower extremities, a previously undetected spinal anomaly may be present. In such patients, it may be prudent to avoid central neuraxial block to prevent neural damage. This instructive case presents such an adult patient in whom previously undiagnosed diastematomyelia, low conus and tethered cord was discovered only in the assessment of postspinal neurological symptoms. PMID- 11493241 TI - Kinin-mediated anaphylactoid reaction implicated in acute intra-operative pulseless electrical activity. AB - A 65-year-old patient undergoing total hip replacement under general anaesthesia suffered acute pulseless electrical activity with a fatal outcome. A kinin mediated analphylactoid reaction following administration of a polygeline plasma expander (Haemaccel) was implicated by in vitro testing. This case report illustrates the diagnostic difficulties posed by non-histaminoid anaphylactoid reactions and the resistance to epinephrine of kinin-mediated hypotension. PMID- 11493242 TI - Adverse reactions to colloids. PMID- 11493243 TI - False-positive epidural catheter aspiration tests in needle through needle combined spinal-epidural anaesthesia. AB - We describe two separate episodes of free aspiration of clear fluid through epidural catheters during needle-through-needle combined spinal-epidural anaesthesia in the lateral position. Although both catheters were assumed to have been located intrathecally, both subsequently gave negative test doses and were used as epidural catheters. In the second patient, the position of the catheter was investigated using contrast spiral computerised tomography scanning, which showed epidural contrast spread only. This is the first time such an occurrence has been recognised, and we consider its significance for catheter site confirmation testing in combined spinal-epidural anaesthesia. PMID- 11493244 TI - Anaesthesia for insertion of bone-anchored hearing aids in children: a 7-year audit. AB - Forty-three children, aged 23 months to 14 years, received 102 anaesthetics for insertion of bone-anchored hearing aids, each lasting approximately 30-60 min. Forty of the children had a recognised syndrome involving the head and neck, including Goldenhar's and Treacher Collin's syndrome. The incidence of congenital heart disease was 19%. Pre-existing conditions, anaesthetic technique, grade of intubation, complications and discharge were audited. Sixteen of the patients were classified as Grade 3 or 4 intubations. Over the 7 years, laryngeal mask airway usage increased for airway maintenance rather than tracheal intubation, as did the use of propofol for induction rather than inhalational methods. Intra operative complications (5.9%) were related to the airway, and postoperative ones (17.6%) mainly to nausea and vomiting. Surgery was performed as a day case in 71% of the patients despite some long-distance travel. PMID- 11493245 TI - Prophylaxis against thromboembolism in patients with traumatic brain injury: a survey of UK practice. AB - Venous thromboembolism is a major complication associated with traumatic brain injury and is responsible for significant morbidity and mortality. There has been a general reluctance over the years to use anticoagulant prophylaxis for patients with head injury who have suffered intracranial bleeding or for whom intracranial surgery is needed. We conducted a postal questionnaire survey of all neurosurgical centres in the United Kingdom, enquiring about the use of thromboprophylactic methods in the management of patients with traumatic brain injury. A diversity of practice and opinion in the use of such methods was evident from the replies received. The survey highlighted concern about the failure to implement even the most simple means of prophylaxis. The evidence for the use of the various methods of prophylaxis is reviewed. PMID- 11493246 TI - A pharmacokinetic model for factor VIII dosing during active haemorrhage in patients with haemophilia A. AB - A theoretical one-compartment pharmacokinetic model is described, the aim of which is to guide factor VIII replacement therapy in patients with haemophilia A suffering from significant and ongoing haemorrhage. Based on our calculations, plasma factor VIII concentrations can decrease at a rapid rate even with non severe blood loss. PMID- 11493247 TI - Extension of epidural blockade in labour for emergency Caesarean section using 2% lidocaine with epinephrine and fentanyl, with or without alkalinisation. AB - In a randomised, double-blind study, we investigated rapid extension of epidural analgesia to surgical anaesthesia for emergency Caesarean section. Parturients receiving epidural analgesia in labour who subsequently required Caesarean section were given a test dose of 3 ml lidocaine 2% with epinephrine 1 : 200 000, followed 3 min later by 12 ml lidocaine 2% with epinephrine 1 : 200 000 and fentanyl 75 microg, to which was added 1.2 ml sodium bicarbonate 8.4% (bicarbonate group; n = 20) or saline (saline group; n = 20). Mean (SD [range]) time to surgical anaesthesia was less in the bicarbonate group (5.2 (1.5) [2-8] min) than the saline group (9.7 (1.6) [6-12] min; mean difference 4.5 min (95% CI 3.5-5.5) min; p < 0.001). Maternal side-effects and neonatal outcome were similar between groups. We conclude that pH-adjusted lidocaine 2% with epinephrine and fentanyl is effective for rapidly establishing surgical anaesthesia in patients with a functioning epidural catheter for labour who require emergency Caesarean section. PMID- 11493248 TI - Prevention and management of hypotension during spinal anaesthesia for elective Caesarean section: a survey of practice. AB - Hypotension during obstetric spinal anaesthesia has traditionally been managed by such measures as fluid preloading, positioning of the patient and the use of vasoconstrictors. However, studies and reports have regularly appeared in the literature disputing the value of conventional management, in particular, the fluid preload. With this in mind, we surveyed UK consultant obstetric anaesthetists to determine current practice in this area. Of the 558 respondents, 486 (87.1%) stated that they routinely give a fluid preload. The fluid chosen by 405 (83.3%) of the preloaders was Hartmann's solution and the usual volume, chosen by 194 (39.9%), was 1000 ml. A simple left lateral position was preferred by 221 respondents (39.6%) overall and in the treatment of hypotension, ephedrine was the sole vasoconstrictor selected by 531 (95.2%). Heavy bupivacaine 0.5% was the local anaesthetic chosen by 545 (97.7%) and 407 (72.9%) respondents indicated the use of additional spinal drugs. PMID- 11493249 TI - Teaching methods. PMID- 11493251 TI - A sticky edge. PMID- 11493252 TI - Hypothermia and rapid, cold, peritoneal crystalloid infusion. PMID- 11493253 TI - Pulse oximeter aided diagnosis of fluid extravasation. PMID- 11493254 TI - Pulse oximetry from a haemofiltration line. PMID- 11493255 TI - Acute epiglottitis, sevoflurane and HIB vaccination. PMID- 11493256 TI - Anaesthesia in pyruvate dehydrogenase deficiency. PMID- 11493257 TI - Sotos' syndrome ellipsis...a rare challenge? PMID- 11493258 TI - Misplaced central venous catheter. PMID- 11493259 TI - Operating theatre exsanguinators pose a low risk for transmission of infection. PMID- 11493260 TI - Three new MHC haplotypes in broiler breeder chickens. AB - Six distinct serotypes of the chicken B blood group system (which encodes the major histocompatibility complex) were identified in a commercial broiler breeder line (Line C). The B serotypes were compared by B-G restriction fragment length polymorphism (RFLP) analysis, allele-specific PCR typing test for B-LBII family genes and nucleotide sequence analysis of expressed B-F and B-LBII family genes. The results indicated the existence of seven distinct B haplotypes. Nucleotide sequence analysis demonstrated that three of the Line C haplotypes encode new B-F and B-LB alleles. PMID- 11493261 TI - Isolation of a porcine UDP-GalNAc transferase cDNA mapping to the region of the blood group EAA locus on pig chromosome 1. AB - In our studies of the genes constituting the porcine A0 blood group system, we have characterized a cDNA, encoding an alpha(1,3)N acetylgalactosaminyltransferase, that putatively represents the blood group A transferase gene. The cDNA has a 1095-bp open reading frame and shares 76.9% nucleotide and 66.7% amino acid identity with the human ABO gene. Using a somatic cell hybrid panel, the cDNA was assigned to the q arm of pig chromosome 1, in the region of the erythrocyte antigen A locus (EAA), which represents the porcine blood group A transferase gene. The RNA corresponding to our cDNA was expressed in the small intestinal mucosae of pigs possessing EAA activity, whereas expression was absent in animals lacking this blood group antigen. The UDP-N acetylgalactosamine (UDP-GalNAc) transferase activity of the gene product, expressed in Chinese hamster ovary (CHO) cells, was specific for the acceptor fucosyl-alpha(1,2)galactopyranoside; the enzyme did not use phenyl-beta-D galactopyranoside (phenyl-beta-D-Gal) as an acceptor. Because the alpha(1,3)GalNAc transferase gene product requires an alpha(1,2)fucosylated acceptor for UDP-GalNAc transferase activity, the alpha(1,2)fucosyltransferase gene product is necessary for the functioning of the alpha(1,3)GalNAc transferase gene product. This mechanism underlies the epistatic effect of the porcine S locus on expression of the blood group A antigen. ABBREVIATIONS: CDS: coding sequence; CHO: Chinese Hamster Ovary; EAA: erythrocyte antigen A; FCS: foetal calf serum; Fucalpha(1,2)Gal: fucosyl-alpha(1,2)galactopyranoside; Gal: galactopyranoside; GGTA1: Galalpha(1,3)Gal transferase; PCR: polymerase chain reaction; phenyl-beta-D-Gal: phenyl-beta-D-galactopyranoside; R: Galbeta1 4Glcbeta1-1Cer; UDP-GalNAc: uridine diphosphate N-acetylgalactosamine PMID- 11493262 TI - Genetic relationship amongst the major non-coding regions of mitochondrial DNAs in wild boars and several breeds of domesticated pigs. AB - We completed phylogenetic analysis of the major non-coding region of the mitochondrial DNA (mtDNA) from 159 animals of eight Euro-American and six East Asian domesticated pig breeds and 164 Japanese and five European wild boars. A total of 62 mtDNA haplotypes were detected. Alignment of these regions revealed nucleotide variations (including gaps) at 73 positions, including 58 sites with transition nucleotide substitutions, and two transversion substitutions. Phylogenetic analysis of the sequences could not organize domestic pig breeds into discrete clusters. In addition, many of the haplotypes found in members of diverged clustering groups were found primarily in Euro-American pig breeds, indicating extensive introgression of Asian domestic pigs into European breeds. Furthermore, phylogenetic analysis allocated the DNA sequences of non-coding regions into two different groups, and the deepest branchpoint of this porcine phylogeny corresponded to 86 000-136 000 years before present. This time of divergence would predate the historical period when the pig is thought to have been domesticated from the wild boar. PMID- 11493263 TI - Chromosome assignment of six dog genes by FISH, and correlation with dog-human Zoo-FISH data. AB - Cross-species chromosome painting analyses have recently demonstrated the presence of regions of conserved synteny between the human and domestic dog genomes, aiding the search for candidate genes for inherited traits. Concerted efforts to subchromosomally assign substantial numbers of dog gene sequences are now needed in order to refine these comparative data, both in terms of marker density and resolution. We have developed novel PCR markers representing three dog genes (ALB, FOS, HNRPA2B1) for which no sequence or mapping data were previously available, to our knowledge. These, in addition to three gene markers previously described (ALDOA, RPE65, VCAM1), were used to isolate and chromosomally assign corresponding large insert genomic clones by fluorescence in situ hybridization (FISH). Chromosome assignments for these six dog genes are discussed in terms of those of the human orthologues, and correlated with existing comparative mapping information, identifying one apparent exception to existing Zoo-FISH data, and aiding refinement of the boundaries of conserved chromosome segments in both genomes. PMID- 11493264 TI - Development and mapping of microsatellites from a microdissected BTA 11-specific DNA library. AB - A chromosome-specific library was developed for Bos taurus autosome 11 by chromosome microdissection and microcloning using a bovine primary fibroblast culture, obtained from a t(X;23) heifer, that spontaneously developed a translocation chromosome involving bovine chromosome 11. The library was screened using (AC)12 oligos, positive clones selected, sequenced and primers developed to generate bovine chromosome 11-specific microsatellite markers. This study suggests that chromosome-specific libraries have great potential for development of microsatellite markers for the construction of marker-saturated linkage maps for each chromosome. PMID- 11493265 TI - DNA cloning and sequence analysis of chicken AFLP. AB - Amplified fragment length polymorphisms (AFLP) have been shown to be useful for linkage mapping in chickens and other domestic animals. It is often desirable to convert AFLP bands to sequence-tagged site (STS) markers, in particular, so that AFLP-based linkage information can be integrated with recombinant DNA clone-based maps. Sixteen chicken AFLP bands were excised from gels, re-amplified, cloned and analysed. All inserts proved to be EcoRI-TaqI fragments, which suggests that unlabelled TaqI-TaqI AFLP fragments do not amplify well, and therefore do not significantly contaminate AFLP bands. For eight of the AFLP, the cloned fragment was used to probe blots of AFLP reaction fingerprints, confirming that the predominant DNA clone indeed contained the polymorphic fragment. Flanking regions of selected AFLP fragments were isolated using Vectorette cloning. The results obtained suggest that the these chicken AFLP most commonly arise from sequence polymorphism at or near the TaqI site. PMID- 11493266 TI - Cytogenetic assignment and genetic characterization of the horse microsatellites, TKY4-18, TKY20, TKY22-24, TKY30-41 derived from a cosmid library. PMID- 11493267 TI - Isolation and characterization of chum salmon microsatellite loci and use across species. PMID- 11493268 TI - A single nucleotide polymorphism in the porcine androgen receptor gene. PMID- 11493269 TI - Characterization of 14 macropod microsatellite genetic markers. PMID- 11493270 TI - Hepatocyte cell lines as the biological component of liver support. PMID- 11493271 TI - Liver stem cells: a potential source of hepatocytes for the treatment of human liver disease. AB - Severe liver injury often leads to the proliferation of oval cells, which differentiate along hepatocytic and biliary lineages. Because oval cells proliferate only when hepatocyte replication is impaired, they are considered to be the progeny of facultative liver stem cells (FLSCs). Identification and isolation of FLSCs has been hampered by the lack of markers that delineate these bipotential progenitors. We hypothesized that transition ductal cells are FLSCs because they are located in a unique anatomical niche sharing tight junctions with a neighboring hepatocyte and another terminal ductular cell. Alternatively, it has been proposed recently that bone marrow-derived stem cells are FLSCs since these cells differentiate along the hepatic lineage following colonization of the liver. The intent of this review is to provide insight into the nature and origin of liver stem cells and to explore the possibility that stem cell technology may lead to the development of clinical modalities for the treatment of human liver disease. PMID- 11493272 TI - Amplification of engrafted hepatocytes by preparative manipulation of the host liver. AB - Scarcity of donor livers is a major obstacle to the general application of hepatocytes for the development of bioartificial liver assist devices as well as intracorporeal engraftment of hepatocytes for the treatment of inherited metabolic diseases. The number of hepatocytes that can be transplanted into the liver safely in a single sitting also limits the utility of this procedure. These limitations could be addressed by providing preferential proliferative advantage to the transplanted cells. Studies using transgenic mouse recipients or donors have indicated that massive repopulation of the host liver by engrafted hepatocytes requires that the transplanted cells are subjected to a proliferative stimulus to which the host hepatocytes cannot respond. Prevention of host hepatocyte proliferation has been achieved by treatment with a plant alkaloid, retrorsine. Because retrorsine is carcinogenic, we have evaluated preparative irradiation for this purpose. The proliferative stimulus may consist of the loss of hepatic mass (e.g., partial hepatectomy, reperfusion injury or induction of Fas-mediated apoptosis by gene transfer) or administration of stimulants of hepatocellular mitosis (e.g., growth factors or thyroid hormone). Potential applications of these preparative manipulations of the host liver include the treatment of inherited metabolic disorders by transplantation of allogeneic hepatocytes, hepatocyte-mediated ex vivo gene therapy, rescuing liver cancer patients from radiation-induced liver damage, and expansion of human hepatocytes in animal livers. PMID- 11493273 TI - Novel strategies for immortalization of human hepatocytes. AB - Normal somatic cells have a finite life span due in part to their inability to maintain telomere length and chromosome stability. Immortalization strategies based on recent advances in telomere biology and aging research have led to the creation of genetically stable, nontumorigenic immortalized cell lines. Reversible immortalization, using the Cre-lox recombination and excision system, has been developed for the expansion of primary cells for cell based clinical therapies. Immortalized human hepatocyte cell lines with differentiated liver functions would find broad applications in biomedical research, especially for pharmacology and toxicology, artificial liver support, and hepatocyte transplantation. The biological basis of these new immortalization methods and their application to human hepatocytes is reviewed. PMID- 11493274 TI - Growth and metabolic activity of immortalized porcine hepatocytes in extracorporeal hollow-fiber liver assist devices. AB - The development of a cell based extracorporeal liver assist device offers a promising clinical approach to bridge individuals suffering from acute liver failure to transplant. However, a major drawback of the existing technology is the lack of a continuous supply of well differentiated hepatocytes. Although some investigators have used primary porcine cells, this approach demands costly, labor-intensive isolation procedures and yields cells with inconsistent detoxification capacity. The limitations of primary cells led us to develop the HepLiu immortalized porcine hepatocyte cell line for use in liver assist devices (LADs). HepLiu cells are nontumorigenic and exhibit multiple hepatic detoxification functions including diazepam and acetaminophen metabolism. To investigate the suitability of HepLiu cells for artificial liver support, morphology, as well as xenobiotic metabolism, was studied in perfused polysulfone hollow-fiber LADs. HepLiu cells were cultured in the intercapillary space of a prototype LAD, and the metabolism of diazepam, acetaminophen, and 7 ethoxycoumarin was evaluated over 25 days in culture. Our results indicated that HepLiu cells proliferated rapidly following inoculation of the LAD until Day 10 when proliferation appeared to cease. Ultrastructural analysis demonstrated that HepLiu cells retained many of the features of primary hepatocytes including desmosomes that sealed bile canalicular-like structures and junctional complexes (intermediate, gap junctions) that appeared concentrated in the paracanalicular areas. Unlike primary porcine hepatocytes, HepLiu cells retained drug metabolic function throughout the 25 day culture period. Diazepam metabolism by HepLiu cells was consistently higher than that of primary cells. Acetaminophen metabolism persisted throughout the 25 day period albeit at a much lower level than the primary cells exhibited on Days 1 or 2. In conclusion, we have shown that HepLiu cells proliferate to occupy the intercapillary space of perfused hollow-fiber LADs following inoculation, and retain their metabolic capacity for Phase I and Phase II detoxification reactions in perfusion culture. Our findings suggest that HepLiu cells may provide an alternative to primary porcine hepatocytes as the cellular component of bioartificial liver support systems. PMID- 11493275 TI - Culture of porcine hepatocytes: the dogma of exogenous matrix revisited. AB - The use of exogenous matrices has been described as an essential component in securing the viability and functionality of hepatocytes in vitro whether cultured for extracorporeal devices or cell transplantation. Here we report on the in vitro culture of porcine hepatocytes in polystyrene tissue-culture flasks without exogenous matrices showing adequate attachment and viability. Cell proliferation was evidenced by uptake of 5-bromo-2'-deoxyuridine, with peaks at Days 2 (19.7 +/ 8.5%), 15 (20.8 +/- 3.3%), and 35 (21.4 +/- 0.3%). Detoxification capacity was assessed by determination of monoethylglycinexylidide, a product of lidocaine metabolism (highest value 156.5 +/- 10.1 ng/ml at Day 4), and by diazepam clearance (maximum clearance 66.2% at Day 6). Diazepam metabolite levels were highest at Day 4 both for temazepam and oxazepam (6.5 +/- 0.1 and 0.10 +/- 0.01, respectively). These results suggest that the need for an exogenous matrix to achieve sustained proliferative activity and differentiated hepatocyte function should not necessarily be considered a sine qua non condition. PMID- 11493276 TI - Monitoring of brain water by chemical shift imaging during ammonia-induced brain swelling in rats after portacaval anastomosis. AB - Brain edema is a leading cause of death in acute liver failure (ALF). In experimental models of ALF, an increase in the content of brain water has been inferred indirectly by measuring intracranial pressure or determined directly via analysis of brain tissue postmortem. In this study, noninvasive proton two dimensional chemical shift imaging (2-D CSI) was used to follow the time course of the development of brain edema in a well characterized model, namely ammonium acetate infusion into rats 48 to 72 h after portacaval anastomosis (PCA). Clear differences between control and experimental rat brains were observed, with an increase of brain water signal only in the parietal cortex of the PCA + ammonia group. Selective swelling of the cerebral cortex points to a cytotoxic mechanism in the evolution of brain edema in this model. CSI signal enhancement was much greater than the gravimetrically determined water content increase. The significantly greater signal change observed with 2-D CSI may reflect enhanced proton density that results from increased water content as well as edema-related alterations in water relaxation times. PMID- 11493277 TI - Extensive in vivo angiogenesis following controlled release of human vascular endothelial cell growth factor: implications for tissue engineering and wound healing. AB - Vascular endothelial cell growth factor (VEGF) has strong stimulating effects on vascularization. Though very potent, VEGF is rapidly degraded due to its short half-life and when administrated by uncontrolled and nonspecific methods; however, its systemic administration in large doses can cause harmful side effects. Controlled release technology would allow delivering desired levels of bioactive VEGF within extended periods and permit examination of the in vivo effects of the compound in a broader way. The objective of this study was to determine the in vitro release behavior of VEGF from calcium alginate microspheres and the potency of this controlled release system in promoting localized neovascularization at the subcutaneous site of the rat model. In vitro release of human VEGF165 (2 and 4 microg/cm3 microsphere) was studied for 3 weeks under static conditions at 25 degrees C, and daily hormone release was measured using a competitive enzyme immunoassay. Following an uncontrolled release within the first 4 days, a quite constant zero-order VEGF release of 50 to 90 and 70 to 120 ng/day was achieved from 2 and 4 microg/cm3 polymer loaded microspheres respectively. In vivo angiogenesis was studied for a period of 8 weeks and evaluated using immunoperoidase staining and histopathological measurements. In vivo studies with rats (n = 24) showed a considerable level of capillary network formation at the epigastric groin fascia of VEGF microsphere-implanted rats starting from the first week. The most extensive neovascularization was observed in the group with 3 week postimplanted 4 microg VEGF containing microspheres; this level of vascularization was quite similar after 8 weeks. While the control group showed no evidence of angiogenesis, the difference in VEGF-induced neovascularization is statistically significant (p < 0.03). Immunostaining of the specimens showed a strong relationship between the release of human VEGF and neovascularization. The controlled VEGF release system described here promotes vigorous angiogenesis and has applicability for tissue engineering and wound healing studies. PMID- 11493278 TI - Bioartificial liver treatment prolongs survival and lowers intracranial pressure in pigs with fulminant hepatic failure. AB - Intracranial hypertension leading to brainstem coning is a major cause of death in fulminant hepatic failure (FHF). We have developed a bioartificial liver (BAL) utilizing plasma perfusion through a bioreactor loaded with porcine hepatocytes and a column with activated charcoal. In a Phase I clinical trial, we observed a decrease in intracranial pressure (ICP) in FHF patients. However, these patients received BAL therapy together with other measures. We therefore examined whether BAL therapy alone could prevent development of intracranial hypertension in pigs with surgically induced FHF. Pigs (40-60 kg) underwent end-to-side portacaval shunt, transection of all hepatic ligaments, and placement of slings around the hepatic artery and bile duct. After 3 days, the slings were tightened to induce liver necrosis. After 4 h, Group 1 pigs (n = 6) underwent a 6 h treatment with the BAL utilizing 10 billion cryopreserved pig hepatocytes and a charcoal column, Group 2 pigs (n = 6) with the BAL containing charcoal but no cells, and Group 3 pigs (n = 6) with the BAL containing neither cells nor charcoal. Group 1 pigs maintained a normal ICP during BAL treatment and for 14 h afterward and because of this effect they survived longer than Groups 2 and 3 animals. In contrast, Groups 2 and 3 pigs showed an early (6-8 h) rise in ICP. PMID- 11493279 TI - In vitro and in vivo evaluation of albumin synthesis rate of porcine hepatocytes in a flat-plate bioreactor. AB - Several configurations of extracorporeal bioartificial liver devices have been developed for the potential treatment of fulminant hepatic failure or as a bridge to liver transplantation. Recently, we developed a microchannel flat-plate bioreactor with an internal membrane oxygenator in which porcine hepatocytes are cultured as a monolayer on the bottom glass surface. In the present study, we investigated synthetic function of porcine hepatocytes in the bioreactor in both in vitro and in vivo flow circuit models. In vitro, albumin synthesis was stable in the bioreactor for up to 4 days of perfusion. In vivo, with the extracorporeal connection of the bioreactor to rat vasculature, porcine albumin was detectable for 24 h in the rat plasma. We also developed a simple mathematical model to predict the in vivo porcine albumin concentration in rat plasma. These results indicate that this configuration of a microchannel flat-plate bioreactor has potential as a liver support device and warrants further investigation. PMID- 11493280 TI - Are standard human coagulation tests suitable in pigs and calves during extracorporeal circulation? AB - The thrombogenicity of membrane oxygenators as well as clotting parameters profiles, using standard human clotting tests, was analyzed in calves and pigs during 6 h perfusion. Three calves and 3 pigs were connected to extracorporeal circulation with standard heparinization. Blood samples were taken for coagulation variables throughout perfusion, and oxygenators were examined for clot deposits at the end of the experiment. Two out of 3 oxygenators of the calf group presented clot deposits while none in the pig group did. Baseline coagulation variables of pigs showed values similar to those of humans while neither extrinsic nor intrinsic pathways could be activated in calves with standard human coagulation tests. The calf model, in conclusion, was confirmed to be a difficult model for the testing of extracorporeal circulation device resistance to thrombus formation, which is, however, not reflected by standard human coagulation tests. The pig model is a better model in which both coagulation pathways could be activated with standard human coagulation tests. PMID- 11493281 TI - In vitro evaluation of the heparin-coated Gyro C1E3 blood pump. AB - The blood-contacting surface, mainly composed of polycarbonate, of the Gyro C1E3 pump was modified using plasma glow discharge to introduce a carboxyl functional group, coated with a base layer of polyethyleneimine as a linker, and coupled with heparin by multi-ionic binding to enhance blood compatibility. A relative surface content of 3.7% sulfur, which demonstrated heparin immobilization on the polycarbonate substrate, was observed on x-ray photoelectron spectroscopy, and a initial bioactivity of approximately 88.5 +/- 7.3 mIU/cm2 was obtained by the chromogenic method for antifactor Xa assay. Furthermore, in vitro observation of platelet and fibrin adherence using bovine blood under dynamic flow conditions for 6 h revealed that the multi-ionically heparinized Gyro C1E3 had significantly stronger antithrombogenecity than the noncoated original type which was evaluated as a good hemocompatible blood pump for clinical use. Not only the Gyro C1E3 but also the ionically heparin-coated Gyro pump are expected to be thromboresistant in clinical use. PMID- 11493282 TI - Dose effect relationship of reviparin in chronic hemodialysis: a crossover study versus nadroparin. AB - Low molecular weight heparins (LMWHs) are used for prevention of clotting in the dialysis circuit. The aim of this trial was to define the optimal dose of a new LMWH and to test the efficiency of a single dose at the start of the session. Fifteen patients were treated according to a double blind and crossover design during 4 blocks of 5 consecutive reviparin doses assigned randomly as 50, 60, 70, 85, and 100 IU anti-Xa/kg. Assessment was carried out on screening of fibrin rings or clots in the arterial and venous air traps and on visual detection of fiber in the dialyzer after rinsing. These clinical results were compared to plasmatic anti-Xa activity and thrombin-antithrombin (TAT) complex generation. A standard dose of 70 IU anti-Xa/kg of nadroparin was used as the control. After a bolus of 50 to 100 IU anti-Xa/kg, the occurrence of fibrin rings and clots in the air traps was dependent on three factors: dose of LMWH, time of the session, and patient status. A bolus of 85 IU anti-Xa/kg of reviparin was effective and safe for sessions of 4 h. For this dose, plasmatic anti-Xa activity was 0.96 +/- 0.28 IU/ml at Hour 2 and 0.82 +/- 0.22 IU/ml at Hour 4. TAT complexes are good markers of the activation of the coagulation. They did not increase during a 4 h session after a reviparin bolus of 100 IU/kg. For the same LMWH dose, the trial shows a great variability of the clinical effect and anti-Xa activities from one patient to another. A single dose of 85 IU anti-Xa/kg of reviparin can be used at the start of the dialysis session as a loading dose. We advise adapting the dose during the subsequent sessions according to the appearance of the blood circuit. The benefit of monitoring anti-Xa activity and TAT complexes could be tested in a further trial. PMID- 11493283 TI - A time for change? PMID- 11493284 TI - On the brink of change? Implications of the review of undergraduate education in New Zealand for mental health nursing. AB - A New Zealand Nursing Council review of undergraduate education provides an ideal opportunity to make much needed changes to the system of preparation for mental health nurses. This article critiques comprehensive nursing education through an examination of its history in New Zealand, recent mental health reports and a projected estimate of workforce needs. Historical analysis reveals a process of marginalization and invisibilization of psychiatric/mental health nursing within comprehensive programmes with a consequent reduction of skills and a weakening of the profession. The author concludes that psychiatric/mental health nursing is a distinct scope of practice which requires specialty undergraduate preparation. PMID- 11493286 TI - Taking neuroleptic medications as the treatment for schizophrenia: A phenomenological study. AB - Research in the area of neuroleptic medications has traditionally relied on quantitative methodologies which view the problem from the researcher's perspective. This phenomenological study was undertaken to explore the experience of taking neuroleptic medications from the individual's perspective using a research approach which has the potential to illuminate and respect participant experiences. Interviews were conducted with 10 individuals who take neuroleptic medications as their main treatment for schizophrenia. Interviews were analysed using an interpretive method which seeks to generate a deeper understanding of the phenomenon under study. The experience of taking neuroleptic medications emerged as incorporating hope for recovery, learning to live with the side effects, embodiment of illness, experiencing issues of control, and struggling to stay well. The results have relevance for practicing and novice mental health nurses, for teachers of undergraduate nursing and allied health students, and carer and consumer groups. PMID- 11493287 TI - Coping with everyday reality: mental health professionals' reflections on the care provided in an acute psychiatric ward. AB - There is little research evidence about how the mental health professionals are coping with the complexity of everyday practice in psychiatric acute care. The aim of this study was to explore mental health professionals' reflections on their work on an acute psychiatric ward. Data were collected using participant observation and interview methods. Three core themes were identified from a qualitative hermeneutic analysis. The first core theme, coping with uncertainty, uncovered a dialectical pattern of the factors contributing to thriving and strain in the working situation. The second core theme, caring for the patient, included the caring process, patients' pathway to acute psychiatric care, as well as the patients' needs and roles on the ward. The third core theme, coping strategies, included five different methods the primary nursing system, concealing versus integrating, milieu therapy, seclusion and the medical orientated model. It was concluded that good mental health care is a result of collaboration between health professionals and the health services. This study highlights the need for support to professionals and for establishing structures that will enable collaboration to take place. Taken together, this may contribute to enhancing the care of the patient and their families. PMID- 11493288 TI - A philosophical analysis of evidence-based practice in mental health nursing. AB - Mental health nurses need to be aware that their knowledge base does not exist in isolation from other cultural practices. They/I/we must become more willing to engage in theoretical problem solving that directly affects clinical practice issues such as the introduction of evidence-based practice. Critical discussion of evidence-based practice should be informed by the complex issues that permeate all our socio-cultural and linguistic practices. This paper examines some of the major philosophical problems in the debate over the use of evidence-based practice in mental health nursing using both Foucault's formulation of discourse analysis and Derrida's construal of deconstruction. The conclusion reached is that postmodern philosophy offers a way to rid nursing of incessant naiive attacks on either quantitative or qualitative research methods which underpin the debate over evidence-based practice in mental health nursing. PMID- 11493289 TI - The relationship between community psychiatric nurses and clients with severe and persistent mental illness: the client's experience. AB - The aim of this phenomenological study was to construct an interpretation of the experience of nurse-patient relationships, in the context of community psychiatric nursing. The purpose of this paper is to focus on the experience of the relationship from the perspective of the clients. Themes of 'having someone looking out for me', 'working in collaboration', and 'being understood and gaining understanding' were identified. This thematic structure was used to understand the meaning of the relationship for the clients. Implications for practice, education, clinical supervision and mental health services are discussed. PMID- 11493290 TI - Unearthing the conflicts between carer and custodian: implications of participation in Section 16 hearings under the Mental Health (Compulsory Assessment and Treatment) Act (1992). AB - Mental health nursing occurs within a legal and ethical framework that presents conflicts between a paternalistic custodial role and an ethical commitment to autonomy. This paper considers the ethical conflict for nurses posed by acting as second health professional at judicial reviews under Section 16 of the Mental Health (Compulsory Treatment and Assessment) Act. Issues of advocacy and paternalism are discussed in light of the conflict between therapeutic and legislated roles. Strategies aimed at protecting the therapeutic relationship are outlined as a response to the current lack of guidelines in this area. Participation in legal processes of committal requires that nurses reflect carefully and critically on the ethical issues raised. PMID- 11493291 TI - Port-wine stain nodules in the adult: report of 20 cases treated by CO2 laser vaporization. AB - BACKGROUND: Port-wine stain (PWS) is a congenital vascular malformation of the superficial dermal vessels. These vessels become progressively ectatic, with development of nodular and hypertrophic areas on the surface of the lesions. OBJECTIVE: To determine the efficacy of CO2 laser vaporization in the treatment of nodules in PWS. METHODS: Twenty adult patients with PWS who developed nodules and hypertrophy on the surface of the lesions were treated by CO2 laser vaporization. Only one pass was performed to each area of the lesions, using a continuous and defocused mode, with a power density of 10 W/cm2. When the treated lesion was very large, several sessions of treatment were necessary to vaporize its entire surface. Photographic controls were performed before and after treatment. RESULTS: The nodules and hypertrophy were removed and the smooth surface of the lesions was reestablished. The color, usually violaceous, was transformed in a slight erythema. CONCLUSION: CO2 laser is a good method for treatment of nodules and hypertrophy in PWS. With adequate precautions, excellent cosmetic results can be obtained. Other more-specific laser systems may be successively used to treat the remaining erythema. PMID- 11493292 TI - Botulinum-A toxin treatment of the lower eyelid improves infraorbital rhytides and widens the eye. AB - Botulinum-A exotoxin (BTX-A) can be used cosmetically to improve rhytides, particularly of the upper one-third of the face. In this study, fifteen women had BTX-A (BOTOX, Allergan, Inc.) injected into the orbicularis oculi muscle. One lower eyelid received two units just subdermally in the midpupillary line three millimeters below the ciliary margin. The opposite periocular area received two units BTX-A in the lower eyelid with 12 units BTX-A injected into the lateral orbital ("crow's foot") area. Three injections of four units each were placed 1.5 cm from the lateral canthus, each 1 cm apart. Patients and physicians independently evaluated the degree of improvement (grade 0 = no improvement, grade 1 = mild improvement, grade 2 = moderate improvement, and grade 3 = dramatic improvement). An independent photographic analysis was performed. Patients reported a grade of 0.73 when two units were injected alone into the lower lid, and a grade of 1.9 when the lower eyelid and the lateral orbital areas were injected. Physician assessment was grade 0.7 with injection of the eyelid alone and grade 1.8 with injection of the lower eyelid and lateral orbital area. Single investigator photographic analysis demonstrated that 40% of the subjects who had injection of the lower eyelid alone had an increased palpebral aperture (IPA), while 86% of the subjects who had injection of the lower eyelid and lateral orbital area had an IPA. Subjects receiving two units alone had an average 0.5 mm IPA and a mean 1.3 mm IPA at full smile. Concomitant treatment of the lateral orbital area produced a mean 1.8 mm IPA at rest and a mean 2.9 mm IPA at full smile. The results were more notable in the Asian eye. Two units of BTX-A injected into the lower eyelid orbicularis oculi muscle improves infraorbital wrinkles, particularly when used in combination with BTX-A treatment of the lateral orbital area. PMID- 11493293 TI - One-pass CO2 versus multiple-pass Er:YAG laser resurfacing in the treatment of rhytides: a comparison side-by-side study of pulsed CO2 and Er:YAG lasers. AB - BACKGROUND: The CO2 laser is normally described as an aggressive resurfacing tool, whereas the erbium:YAG laser has enjoyed a reputation as the ideal tool for superficial resurfacing. The implication from many studies is that the CO2 laser is incapable of "minimally invasive" resurfacing. OBJECTIVE: To compare a short pulsed CO2 laser with an Er:YAG laser over a range of parameters intended to produce equivalent microscopic and clinical injuries. METHODS: A prospective, randomized, comparative interventional trial was conducted in a tertiary care teaching hospital. Thirteen patients with facial wrinkles were enrolled in the study. A side-by-side comparison was performed using periorbital and perioral regions as treatment sites. One side was treated with a pulsed CO2 laser and the other with an Er:YAG laser. Postauricular skin was treated in an identical fashion to the study sites and biopsied for microscopic analysis. The biopsies were obtained before treatment, immediately after treatment, and either 3 or 6 months after treatment to evaluate the acute level of injury and subsequent degree of fibroplasia. Photographs were taken at baseline, immediately after treatment, 1, 2, and 6 weeks, and 3 and 6 months after treatment. Nine physicians evaluated the photographs for erythema, pigmentation, and wrinkle improvement. RESULTS: Investigator assessment showed no statistically significant differences between the lasers with respect to hyperpigmentation and wrinkle reduction. There was less erythema at the CO2 laser-treated sites 2 weeks after treatment; the differences had resolved by 6 weeks after treatment. Histologic examination demonstrated equivalent dermal thermal injury on immediate postoperative biopsies and equivalent fibroplasia on subsequent biopsies. Both CO2 and Er:YAG laser treated sites showed overall modest wrinkle improvement compared to the pretreatment photographs. CONCLUSION: When CO2 and Er:YAG lasers are used in a manner such that there are equivalent immediate postoperative histologic results, equivalent healing and cosmetic improvement occurs. One can use CO2 laser with one pass to mimic a moderately aggressive Er:YAG laser treatment. PMID- 11493294 TI - Evaluation of survival rate after follicular unit transplantation using the KNU implanter. AB - BACKGROUND: Many hair transplant surgeons have advocated the use of micrograft megasessions for the purpose of a more natural looking end result, especially in the Oriental with higher skin/hair color contrast and darker, coarse, straight hairs. But it also has some fundamental limitations. Most important are a low graft yield and a low density after transplantation. There are several reports about the survival rate of micrografts in Caucasians, though the results are variable, but few reports about the survival rate and fate of micrografts in Orientals. OBJECTIVE: To compare the survival rate of one-hair follicular units with that of two-hair follicular units using the KNU implanter, to ascertain the average survival rate of micrografts (one- and two-hair follicular units), and to evaluate the fate of grafted hair according to time. METHODS: Two templates of 1.5 cm2 were made by tattooing on both sides of the frontoparietal recess areas in 11 patients with male pattern baldness (beyond Norwood type IIIa). The authors planted one-hair unit micrografts in the left template and two-hair unit micrografts in the right template, which were prepared by the concept of follicular unit, and counted the surviving number of follicular units at 1 and 3 months and total hairs at 6 and 12 months after transplantation. RESULTS: The mean survival rate by the number of follicular units was 47.3 and 57.4% at 1 and 3 months after transplantation, respectively. The mean survival rate by the total number of hairs was 92.0 and 90.4% at 6 and 12 months after transplantation, respectively. CONCLUSION: About 50% of the transplanted hairs fell out in 1 month, but at 6 months the survival rate of follicular unit transplantation using the KNU implanter showed a good result (92%), and there were no significant differences in the mean survival rate of follicular unit and total hairs between one-hair and two-hair units at 1, 3, 6, and 12 months after transplantation. PMID- 11493295 TI - Silicone versus nonsilicone gel dressings: a controlled trial. AB - BACKGROUND: Silicone gel dressings decrease scar volume and soften hypertrophic tissue, allowing it to be more easily controlled by other methods. Although silicone does not appear to be an essential component of the treatment, nonsilicone dressings have been reported to cause no change in physical parameters during a 2-month treatment period. OBJECTIVE: To compare silicone and nonsilicone gel dressings in the treatment of keloids and hypertrophic scars, including a control group, and to evaluate the effectiveness of these treatments using two new assessment techniques. METHODS: Patients were randomly chosen to receive silicone or nonsilicone gel dressings in a 4.5-month controlled prospective study. Scar size, induration, and symptoms were evaluated before and after the treatment. Scar color was visually measured using a color palette catalog, and a new device was developed to measure intracicatricial pressure. RESULTS: All of the measured parameters were significantly reduced in both silicone- and nonsilicone-treated groups, as compared to the control, with no significant differences between them. CONCLUSION: Silicone and nonsilicone gel dressings are equally effective in the treatment of keloids and hypertrophic scars. PMID- 11493296 TI - Er:YAG laser resurfacing using combined ablation and coagulation modes. AB - BACKGROUND: The two main laser types used in resurfacing, the CO2 and the Er:YAG lasers, have their supporters and detractors, and each system has clear advantages and disadvantages. OBJECTIVE: The Er:YAG laser can be used in the usual efficient ablative mode, followed by reprogramming to achieve nonablative deeper dermal coagulation associated with the CO2 laser, thereby achieving the main advantages of both laser types. PATIENTS AND METHODS: Twenty-three female patients, ages 42-72 yrs, skin types I-IV, were treated. The epidermis was first removed in the ablative settings in a single pass with 50% overlap. The Er:YAG laser was reprogrammed for the subablative mode, and several passes produced controlled residual thermal damage (RTD) without further ablation. RESULTS: At 2 months postresurfacing the results were assessed. Thirteen patients were rated "very good," eight as "good," and two as "fair." CONCLUSION: The dual mode Er:YAG laser can first be used in the ablative mode to remove the epidermis with minimal RTD, following which, in the subablative mode, the same laser induces a controlled layer of dermal RTD, stimulating the dermis to achieve collagenesis and collagen remodeling and giving good long-term results. PMID- 11493297 TI - The efficacy of powered liposuction. AB - BACKGROUND: Powered liposuction is a relatively new innovation for more efficient removal of adipose tissue. OBJECTIVE: To evaluate the effectiveness of powered liposuction in removing adipose tissue when compared to traditional liposuction. METHODS: Four powered liposuction devices were evaluated in the power on mode vs. the power off. The fat extracted in each of these modes was measured in a mucous specimen trap. RESULTS: There was increased fat extraction in the powered mode for all instruments. The increased rate of fat extraction varied from 20 to 45% between instruments. the overall increased extraction in powered vs. nonpowered mode was 30%. CONCLUSION: The powered liposuction devices tested significantly increase the efficacy of subcutaneous fat removal during liposuction. PMID- 11493298 TI - Elliptografting: the right fit. AB - BACKGROUND: A number of new recipient grafting techniques have evolved employing micrografting, minigrafting, and follicular unit technologies as larger hair transplantation sessions incorporating smaller grafts have evolved. OBJECTIVE: To introduce a new recipient graft device-the elliptograft-and compare it to standard circular minigrafts and laser-generated slot grafts. METHODS: A blinded study was performed of 30 patients (mean age 44 years) with grades II-V male pattern Norwood or medium Ludwig female pattern androgenetic alopecia. Ten patients were transplanted with the second-generation elliptograft punch, 10 were treated with 2 mm circular minigrafts, and 10 were transplanted with the recipient sites created by the hybrid Er:YAG/CO2 laser in a slot configuration. The front three rows in each patient were transplanted utilizing No-Kor needle generated micrografts in all three patient study subgroups. RESULTS: Comparable hair growth density was achieved utilizing conventional circular minigraft, freehand laser slit, and elliptograft technologies. Healing time was slightly prolonged in the laser transplant subgroup; however, the time for initial hair growth was comparable in all three patient populations. Aesthetic improvement was judged superior in the elliptograft population by blinded physician observers, which correlated with good patient satisfaction. CONCLUSION: Combined micro-mini elliptografting utilizing the newly described elliptograft punch produces excellent hair density correlated with high patient satisfaction. PMID- 11493299 TI - Histologic evaluation of a Q-switched Nd:YAG laser in the nonablative treatment of wrinkles. AB - BACKGROUND: A variety of nonablative dermal remodeling techniques are currently available. The Q-switched Nd:YAG laser, in addition to its role in tattoo removal, hair removal, and vascular and pigmented lesion treatment, may also play a role in dermal remodeling for the treatment of wrinkles. The histologic changes seen in human skin after Q-switched Nd:YAG laser exposure have yet to be evaluated. OBJECTIVE: To study histologic changes after the use of a Q-switched Nd:YAG laser in the nonablative treatment of photoaged skin. METHODS: Sun-damaged infrauricular skin from six female subjects was exposed to a Q-switched Nd:YAG laser at fluences of 7 J/cm2 with two laser passes. Histologic examinations were performed before laser treatment and 3 months later. RESULTS: Biopsy specimens showed slight fibrosis in the superficial papillary dermis with unremarkable epidermal changes. CONCLUSION: The Q-switched Nd:YAG laser produced morphologic changes similar to, but lesser in degree, than those seen with both CO2 and Er:YAG laser resurfacing. PMID- 11493300 TI - The Webster-type face and neck lift: an extensive cervico-facial rhytidectomy employing a minimally invasive technique. AB - BACKGROUND: Facelifting has incorporated a wide range of new techniques and developments over the past few decades. Many techniques employ a more aggressive surgical approach that may be advocated for seeming advantage, yet upon closer review may be unwarranted for the average patient seeking facial rejuvenation. The Webster-type face and neck lift utilizes a minimally invasive approach to thoroughly resupport the face and neck substructure and skin. This approach affords the patient excellent results while limiting untoward complications. OBJECTIVE: To evaluate the efficacy of the Webster-type face and neck lift. METHODS: The Webster-type face and neck lift was utilized in 200 facelift patients. We reviewed these cases for aesthetic results and associated complications. RESULTS: The great majority of patients have been uniformly pleased with the aesthetic restoration via the Webster-type lift. Postoperative courses have been routinely uneventful and major complications have been avoided. CONCLUSION: The evolution of more invasive approaches to cervicofacial rhytidectomy has not been shown to produce consistently better or longer-lasting results. Many of these aggressive techniques have extended the operating time, heightened the potential morbidity of the operation, and prolonged the duration of convalescence. The Webster-type lift is a proven, modern approach that provides comparable and lasting results while limiting risk and avoiding serious complications. PMID- 11493301 TI - Cutaneous surgery in patients receiving warfarin therapy. AB - BACKGROUND: Anticoagulant treatment with warfarin is an essential therapy in patients with prosthetic heart valves and atrial fibrillation to prevent thromboembolisms. The question whether to stop warfarin treatment in patients undergoing cutaneous surgery is debatable. OBJECTIVE: To evaluate the outcome of surgery in patients that were treated with warfarin and underwent excisional and Mohs surgeries. METHODS: Warfarin therapy was continued in all patients that underwent excisional and Mohs surgery in our practice from November 1999 to November 2000. Perioperative complications such as bleeding and cosmetic outcome are evaluated. RESULTS: A total of 560 patients underwent Mohs surgery and 530 patients underwent excisional surgery. Sixteen patients (1.5%) were treated with coumadin with international normalized ratio (INR) values within the therapeutic values. Seventy-seven patients that underwent surgery on the same days as the warfarin-treated patients served as the control group. Intraoperative bleeding was easily controlled and postoperative bleeding was not recorded in any of the patients. All wounds healed without any complication, including full-thickness grafts. CONCLUSION: Coumadin treatment should be continued in patients undergoing cutaneous surgery. This will decrease the risk of thromboembolic events. PMID- 11493302 TI - The impact of herbal medicines on dermatologic surgery. AB - BACKGROUND: In recent years herbal medicines and supplements have become increasingly popular. With their increased popularity, more publications are warning about the potential harmful effects of some of these products. OBJECTIVE: To present scientific evidence of the benefits and surgical risks of herbal products. METHODS: A Medline search and review of the literature was performed. RESULTS: Many herbal medicines are relevant in dermatologic surgery since Ginkgo biloba, garlic, ginger, ginseng, feverfew, and vitamin E may increase the risk of bleeding, and ephedra may potentiate the side effects of epinephrine. CONCLUSION: Dermatologists should be aware of these herbal products and their uses. Many of these products prescribed by alternative medicine physicians or purchased over the counter should be discontinued prior to dermatologic surgery to minimize the risk of surgical complications. PMID- 11493303 TI - A painless subungual osteoid osteoma. AB - BACKGROUND: Osteoid osteoma is a benign bone tumor. Its etiology is not fully understood and the role of trauma is still elusive. OBJECTIVE: Osteoid osteoma mostly presents with a poorly localized pain that is worst at night and characteristically relieved by salicylates. It usually occurs on the weight bearing bones of the lower extremities, but toe location is quite rare. Here, we present a case of painless osteoid osteoma located subungually on the dorsum of the great toe. RESULT: A 29-year-old woman presented with a painless subungual mass on the dorsum of her great toe. Subungual exotosis, osteochondroma, and osteoma were considered in the differential diagnosis and the lesion was totally excised. Histopathologic examination showed characteristic findings of osteoid osteoma. CONCLUSION: A painless osteoid osteoma is rarely seen and it can be easily misdiagnosed if it occurs in an atypical location such as the subungual area. PMID- 11493304 TI - Erosive pustular dermatosis of the scalp after skin grafting. AB - Erosive pustular dermatosis of the scalp is a rare condition of unknown etiology that usually occurs in the elderly and is characterized by pustules that appear on the scalp leading to scarring alopecia. The histopathology is not specific. Its onset has been related with previous trauma on the scalp. Only three cases after skin grafting have been reported. We describe a case of erosive pustular dermatosis of the scalp appearing on a split-thickness skin graft placed after excision of a basal cell carcinoma. PMID- 11493305 TI - Mohs micrographic surgery of a plexiform fibrohistiocytic tumor. AB - BACKGROUND: Plexiform fibrohistiocytic tumor (PFT) is a rare mesenchymal neoplasm first described by Enzinger and Zhang in 1988. Clinically it is characterized by slow growth, frequent local recurrences, and rare systemic metastasis. These tumors occur chiefly in children and young adults and are most commonly located on the shoulders and forearms. OBJECTIVE: To present a case report of an incompletely excised PFT, its complete resection using simple excision and Mohs micrographic surgery, and review of the literature. METHODS: An 11-year-old Hispanic girl was evaluated for the treatment of an incompletely excised plexiform fibrohistiocytic neoplasm located in the right axilla. Mohs micrographic surgery (MMS) was chosen because of the ill-defined borders and the need for tissue conservation. The patient underwent a two-stage, six section, micrographically controlled excision. Upon completion of the MMS a 2 mm final stage, taken as a peripheral and deep section around and underneath the cleared area, was submitted for paraffin embedding. Residual plexiform histiocytic tumor was found at the margin of resection in one location. Reevaluation of the Mohs slides demonstrated the possible presence of tumor at one deep focus. The tumor was reexcised with a 5 mm margin and repeat hematoxylin and eosin staining showed no residual tumor. There has been no evidence of recurrence in 4 years. RESULTS: Complete resection of the PFT and absence of tumor recurrence 4 years later. CONCLUSION: Over the past few years the list of neoplasms for which MMS is the treatment of choice has steadily grown. PFT is a recently described locally aggressive mesenchymal neoplasm with potential for distant metastasis. To our knowledge this is the first time MMS has been used to resect this tumor. To ensure the chance of complete extirpation we recommend the harvest of an additional stage for hematoxylin and eosin staining, as is done in some aggressive squamous cell carcinomas. PMID- 11493306 TI - Granular cell tumor treated with Mohs micrographic surgery: report of a case and review of the literature. AB - BACKGROUND: Granular cell tumors are uncommon soft tissue neoplasms of neural origin that most often arise in the oral cavity. Penile lesions are distinctly uncommon. Mohs micrographic surgery (MMS) is a highly effective treatment for several cutaneous neoplasms when tissue conservation is crucial. OBJECTIVE: To examine the use of MMS for this soft tissue malignancy. METHODS: We report only the eighth granular cell tumor of the penis. The literature was reviewed regarding the use of MMS for these neoplasms. RESULTS: MMS was performed to minimize the destruction of normal tissue in treating this patient's granular cell tumor. Only one previous article documented the use of MMS for this soft tissue tumor. CONCLUSION: Although utilized infrequently to treat granular cell tumors, MMS may prove beneficial when lesion location or size render tissue conservation or assuredness of cure paramount. PMID- 11493307 TI - Reconstruction of the lower lip. PMID- 11493308 TI - A simple device for incision retraction and protection in endoscopic-assisted brow and forehead lifting. AB - BACKGROUND: Endoscopic-assisted cosmetic surgery has revolutionized various procedures. Forehead and brow lifting performed with endoscopic technique has been shown to be predictable and has fewer complications than open techniques. Providing surgical access and protecting the hair follicles is paramount in endoscopically assisted brow and forehead lifting. OBJECTIVE: To describe a simple retraction device to assist in incision retraction and protect hair follicles. METHODS: A simple, inexpensive retraction device is described that has been used in 60 endoscopic brow incisions to effectively protect the hair follicles and retract incisions for operative techniques. In addition, other methods of follicular protection are discussed. RESULTS: Decreased incisional alopecia and improved surgical access are provided by the use of a simple retraction device and attention to follicular preservation. CONCLUSION: Endoscopic-assisted brow and forehead lifting is becoming the preferred method of upper facial rejuvenation. There is a steep learning curve and often the lack of attention to hair follicle protection results in localized incisional alopecia. In addition, improper surgical access complicates the procedure. A simple device is described to assist in retraction and follicular preservation. PMID- 11493309 TI - Short and long-term follow-up of nonablative 1320 nm Nd:YAG laser facial rejuvenation. PMID- 11493310 TI - Molecular genetics of pseudoxanthoma elasticum. AB - Pseudoxanthoma elasticum (PXE), a systemic heritable connective tissue disorder, is characterized by progressive calcification of elastic structures in the skin, the eyes and the cardiovascular system, with considerable intra- and interfamilial phenotypic variability. Recently, underlying genetic defects have been identified in the ABCC6 gene, which resides on the chromosomal locus 16p13.1 and encodes the MRP6 protein, a member of the ATP-binding cassette (ABC) family of proteins. The affected individuals are homozygous or compound heterozygous for a spectrum of genetic lesions, including nonsense and missense mutations, or deletions and splice-site alterations, confirming the autosomal recessive nature of this condition. Analysis of the deduced primary sequence suggests that MRP6 is a transmembrane transporter, but its function has not been delineated yet. Surprisingly, however, MRP6 is expressed primarily, if not exclusively, in the liver and the kidneys, suggesting that PXE may be a primary metabolic disorder with secondary involvement of elastic fibers. Identification of mutations in the ABCC6 gene in PXE provides a means for prenatal and presymptomatic testing in families at risk for recurrence. DNA-based analyses will also identify heterozygous carriers who may be at risk for development of limited manifestations of the disease as a result of compounding genetic factors and/or environmental modifiers. PMID- 11493311 TI - Trans-species hair growth induction by human hair follicle dermal papillae. AB - A series of experimental bioassays has shown that the dermal papilla of the adult rodent vibrissa hair follicle retains unique inductive properties. In view of the many phenotypic and functional differences between specific hair follicle types, and the growing interest in hair follicle biology and disease, it remains important to establish that the human hair follicle dermal papilla has equivalent capabilities. In this study we tested the ability of human hair follicle papillae to induce hair growth when implanted into transected, athymic mouse vibrissa follicles. The implanted papillae that interacted with mouse follicle epithelium created new fibre-producing follicle end bulbs. The origin of the papillae in the recombinant structures was confirmed using laser capture microdissection and human specific gender determination by PCR. The demonstration that intact adult human dermal papillae can induce hair growth has implications for molecular analysis of basic hair growth mechanisms, particularly since the study involved common epithelial-mesenchymal signalling and recognition properties across species. It also improves the prospects for a cell-based clinical approach to hair follicle disorders. PMID- 11493312 TI - Psoriasis scales contain C5a as the predominant chemotaxin for monocyte-derived dendritic cells. AB - Dendritic cells seem to be of major importance for the pathogenesis of psoriasis. They are increased in number in lesional psoriatic skin which is thought to be due to an increased influx from the peripheral blood regulated by chemotaxins. Using a biological/biochemical approach we have addressed the question whether psoriasis scale extracts contain proteinaceous chemotaxins for dendritic cells. Human monocytes differentiated into dendritic cells by culture with GM-CSF and IL 4 (MoDC) served as responder cells. Chemotactic activity for MoDC was purified by several HPLC-steps. The results of our study show that C5a/C5adesarg is the major chemotactic peptide for MoDC in psoriasis scale extracts. In comparison to other stimuli such as fMLP or monocyte chemotactic peptide 1 (MCP-1) C5a proved to be a most potent and efficient chemotaxin for MoDC. C5a co-eluted with MRP14/calgranulin B which is present in large amounts in psoriasis scale extracts as identified by amino acid sequencing. However, MRP14/calgranulin B did not possess any chemotactic activity for MoDC. Our results provide evidence that C5a/C5adesarg although not specific for dendritic cells seems to be the major chemoattractant for these cells in lesional psoriasis skin. PMID- 11493313 TI - Release of soluble tryptase but only minor amounts of chymase activity from cutaneous mast cells. AB - Tryptase and chymase are the major serine proteinases of skin mast cells but their biologic significance depends on their activity. In this study, we demonstrate the release of soluble activity of tryptase, but not markedly that of chymase, into skin blister fluids induced by freezing with liquid nitrogen as well as into supernatant during incubation of 8 whole skin specimens with compound 48/80 for up to 2 days followed by sonication. Incubation of 3 other skin specimens in compound 48/80 for up to 2 days revealed that the number of mast cells displaying tryptase activity decreased significantly on day 2, and the number of mast cells showing chymase activity (but not those showing chymase immunoreactivity) decreased significantly on day 1 but not thereafter on day 2. The results of 3 skin organ cultures for up to 14 days showed steady decrease in the number of tryptase-positive cells but persistence of mast cells containing chymase activity. Chymase in solution was sensitively inhibited by 0.01 mg/ml alpha1-antichymotrypsin but higher concentrations (0.3-3.0 mg/ml) were needed for inhibiting chymase on skin sections. In conclusion, after mast cell degranulation tryptase activity is substantially solubilized and it may potentially affect both local and distant skin structures. Instead, chymase is partially inactivated and the remaining chymase activity persists at the site of degranulation having only local effects. PMID- 11493314 TI - Bullous pemphigoid sera react specifically with various domains of BP230, most frequently with C-terminal domain, by immunoblot analyses using bacterial recombinant proteins covering the entire molecule. AB - By immunoblot analyses of normal human epidermal extracts, the 230 kDa bullous pemphigoid antigen (BP230) is recognized by most bullous pemphigoid sera. By polymerase chain reaction using keratinocyte cDNA library as a template, we successfully amplified 3 cDNAs of about 3 kb, which covered whole human BP230 molecule. By inserting the cDNAs into bacterial expression vector pGEX, we prepared 3 different recombinant glutathione-S-transferase-fusion proteins, which roughly presented N-terminal domain, central rod domain and C-terminal domain of BP230. By immunoblotting using these 3 recombinant proteins, we demonstrated that the majority of bullous pemphigoid sera reacted clearly with multiple recombinant proteins of BP230, most frequently with C-terminal domain. We also examined sera of pemphigus vulgaris, pemphigus foliaceus and herpetiform pemphigus that showed BP230-like protein band by immunoblotting of epidermal extracts, as well as paraneoplastic pemphigus, for reactivity with the 3 recombinant proteins. In the study, we found that only very few of these non-bullous pemphigoid sera reacted with some of the recombinant proteins. These results indicate that the BP230 is specifically reacted by bullous pemphigoid sera, and that the immunoblotting using the BP230 recombinant proteins should be a useful tool for the diagnosis of bullous pemphigoid. PMID- 11493315 TI - An improved and rapid method to construct skin equivalents from human hair follicles and fibroblasts. AB - To produce sufficient amounts of high quality skin equivalents (SE), either allogenic for dermatopharmacological and dermatotoxicological studies or autologous for transplantation purposes, we established a rapid, easy and cost effective three-dimensional SE model on the basis of human dermal fibroblasts, collagen and freshly plucked hair follicles. Acidic liquid collagen was polymerized with sodium hydroxide in the presence of fibroblasts to form a dermal equivalent (DE) resembling normal human dermis. At 24 h later, freshly plucked hair follicles were implanted into the surface of these DEs after cutting their bulbs off. Another 48 h later, the surface of the SEs was lifted to the air liquid interface. Fourteen days after implantation, outgrowing keratinocytes from the outer root sheath of the hair follicles completely covered the surface of the SE and built a fully developed, multi-layered and cornified epidermis. Histology and immunofluorescence studies with specific antibodies directed against components of keratinocytes, fibroblasts, cell-adhesion molecules, different extracellular matrix and basement membrane proteins revealed the similarity of our three-dimensional SEs to the in vivo situation in normal human skin. Using autologous cell sources and cell culture media enriched with serum from the respective cell donor, it will be possible to use these SEs for autologous transplantation, thereby reducing the risk of transplant rejection. PMID- 11493316 TI - Solar simulated irradiation modulates gene expression and activity of antioxidant enzymes in cultured human dermal fibroblasts. AB - Exposure of skin to solar irradiation generates reactive oxygen species that damage DNA, membranes, mitochondria and proteins. To protect against such damage, skin cells have evolved antioxidant enzymes including glutathione peroxidase (GSH Px), copper and zinc-dependent superoxide dismutase (SOD1), the mitochondrial manganese-dependent superoxide dismutase (SOD2), and catalase. This report examines the effect of a single low or moderate dose exposure to solar-simulating combined UVB and UVA irradiation on the gene expression and activities of these antioxidant enzymes in cultured normal human fibroblasts. We find that both doses initially decrease GSH-Px, SOD2 and catalase activities, but within 5 days after irradiation the activities of the enzymes return to pre-irradiation level (catalase) or are induced slightly (SOD1, GSH-Px) or substantially (SOD2) above the basal level. For SOD1, SOD2 and catalase, the higher dose also detectably modulates the mRNA level of these enzymes. Our results indicate that the effects of a single physiologic solar simulated irradiation dose persist for at least several days and suggest that skin cells prepare for subsequent exposure to damaging irradiation by upregulating this antioxidant defense system, in particular the mitochondrial SOD2. Our findings are consistent with the existence of a broad-based SOS-like response in irradiated human skin. PMID- 11493317 TI - Early migration of cutaneous lymphocyte-associated antigen (CLA) positive T cells into evolving psoriatic plaques. AB - The majority of T cells in lesional psoriatic skin express the skin homing receptor, cutaneous lymphocyte-associated antigen (CLA). We investigated whether this reflects the selective migration of CLA positive cells into evolving psoriatic plaques, consistent with an important role in disease onset, or whether this occurs in the context of an established cutaneous inflammatory response. We identified the advancing edge of plaques in 16 patients with chronic plaque psoriasis using scanning laser Doppler fluxmetry, and performed immunohistochemical analysis of i) lesional psoriatic skin, ii) clinically normal skin immediately in front of the advancing plaque edge, and iii) uninvolved skin distant from the plaque edge. The T-cell infiltrate was characterized using monoclonal antibodies to CD3, CLA and the integrin alphaEbeta7, which is associated with the retention of lymphocytes at mucosal sites. Epithelial proliferation was assessed using a monoclonal antibody to the nuclear proliferation marker Ki67. There was enrichment of CLA positive T cells in evolving psoriatic skin compared to distant, uninvolved skin (mean CLA positive 75.9% vs 47.8%; P<0.003). This accumulation of CLA positive cells occurred before epidermal hyperproliferation was evident, suggesting that this population of cells plays an important, early role in disease pathogenesis. Established lesional psoriatic skin contained a mixed infiltrate of CLA positive (mean 53.2%) and alphaEbeta7 positive (mean 18.2%) cells, suggesting less tissue-specific T cell infiltration, although an additional, specific role for alphaEbeta7 in cutaneous inflammation cannot be excluded. Furthermore, this study has highlighted scanning laser Doppler fluxmetry as a useful investigative tool, permitting analysis of the earliest and therefore potentially most important changes in psoriatic plaque formation. PMID- 11493318 TI - Identification of a novel mutation in 3beta-hydroxysteroid-Delta8-Delta7 isomerase in a case of Conradi-Hunermann-Happle syndrome. AB - The X-linked dominant Conradi-Hunermann-Happle (CDPX2, MIM 302960) syndrome belongs to the rare, heterogeneous group of diseases called chondrodysplasia punctata. The disease has been connected recently with deficiency of 3beta hydroxysteroid-Delta8-Delta7-isomerase (also called emopamil-binding protein, EBP), catalysing an intermediate step in the conversion of lanosterol to cholesterol (1, 2). We report a case of CDPX2 with a new missense mutation (C-->G 439) in exon 4, leading to a R147G aminoacid substitution in the EBP. PMID- 11493319 TI - Ought patients to follow professional advice? PMID- 11493320 TI - Healthcare in a land called PeoplePower: nothing about me without me. AB - In a 5-day retreat at a Salzburg Seminar attended by 64 individuals from 29 countries, teams of health professionals, patient advocates, artists, reporters and social scientists adopted the guiding principle of 'nothing about me without me' and created the country of PeoplePower. Designed to shift health care from 'biomedicine' to 'infomedicine', patients and health workers throughout PeoplePower join in informed, shared decision-making and governance. Drawing, where possible, on computer-based guidance and communication technologies, patients and clinicians contribute actively to the patient record, transcripts of clinical encounters are shared, and patient education occurs primarily in the home, school and community-based organizations. Patients and clinicians jointly develop individual 'quality contracts', serving as building blocks for quality measurement and improvement systems that aggregate data, while reflecting unique attributes of individual patients and clinicians. Patients donate process and outcome data to national data banks that fuel epidemiological research and evidence-based improvement systems. In PeoplePower hospitals, constant patient and employee feedback informs quality improvement work teams of patients and health professionals. Volunteers work actively in all units, patient rooms are information centres that transform their shape and decor as needs and individual preferences dictate, and arts and humanities programmes nourish the spirit. In the community, from the earliest school days the citizenry works with health professionals to adopt responsible health behaviours. Communities join in selecting and educating health professionals and barter systems improve access to care. Finally, lay individuals partner with professionals on all local, regional and national governmental and private health agencies. PMID- 11493321 TI - Consumers' views of quality in the consultation and their relevance to 'shared decision-making' approaches. AB - BACKGROUND: There is a recognized need to assess the effects of shared decision making and other communication interventions. However, the outcomes usually assessed for evidence of 'effectiveness' are determined by researchers and have not been based on consumers' views. AIM: This study aimed to identify the important outcomes of consultations for consumers, and to compare with those reported in the current literature. SETTING AND PARTICIPANTS: Forty-seven participants attending six focus group interviews. Most interviews took place in and all were orientated towards the UK primary care setting. METHODS: Focus group study. RESULTS: Many affective outcomes were identified, consistent with the current literature trends. However, many cognitive and behavioural outcomes that are assessed in the current literature were not noted by participants as important. Furthermore, a broader range of outcomes than is evident in the current literature was viewed as important to these participants. CONCLUSIONS: There is a need to revisit the outcomes which are assessed in decision-making and communication research. The outcomes of greatest importance to consumers must be identified and confirmed by new research which is based directly on the views of consumers themselves. PMID- 11493323 TI - A 'Third Way' for lay involvement: what evidence so far? AB - AIMS AND OBJECTIVES: This article considers evidence regarding lay involvement in the NHS, following the White Paper's commitment to rebuild public confidence in an NHS 'accountable to patients and open to the public and shaped by their views'. It looks at two aspects of lay involvement: the lay board member's involvement in primary care group (PCG) decision-making and the engagement of the PCG with the wider public. METHODS: The paper analyses data from the first sweep of the annual Tracker Survey of a sample of PCGs in England, led by the National Primary Care Research and Development Centre in collaboration with the King's Fund between September and December 1999. It draws specifically from the postal questionnaires sent to lay members. Firstly, however, it contextualizes this data by reviewing the history of lay involvement before 1997 in the NHS and particularly in primary care. CONCLUSIONS: The paper concludes that, during the first 6 months of their operation, the lay voice was faintly heard in PCGs. The lay member's role in decision-making at board-level was peripheral. The majority rated their involvement in key aspects of decision-making as low and their influence on decision-making below that of other board members including the Chief Officer, the chair and the GP board members. Beyond the arena of the board, what little contact there was with the lay voice has taken the shape of informing rather than consulting. Mitigating factors include the early stage at which the survey was completed and the lack of precedents for lay involvement in primary care in a broad sense on which PCGs can draw. PMID- 11493324 TI - Eliciting patients' values by use of 'willingness to pay': letting the theory drive the method. AB - OBJECTIVE: To describe the three different ways in which 'willingness to pay' (WTP) has been used to elicit patients' values of alternative interventions. DESIGN: For each of the three approaches a survey of patients or the public was undertaken. SETTING, PARTICIPANTS AND INTERVENTIONS: studied For two surveys, the setting was Aberdeen Maternity Hospital, where pregnant women were asked about their WTP for different methods of prenatal screening for cystic fibrosis. In the third survey, parents of primary and secondary schoolchildren were asked about their WTP for different ways of providing child health services. MAIN OUTCOME MEASURES: Ability of WTP to discriminate between options (i.e. to say whether one option is 'better' than another) and the consistency of WTP with stated preferences. RESULTS: Experience with some methods shows that, despite the apparent logic of the technique, it is difficult to elicit consistent responses whereby WTP values derived match the rankings of interventions compared. The most promising technique, the 'marginal approach', happens to conform more with economic theory than other approaches. Potential limitations of WTP, such as its association with ability to pay, are discussed, as are approaches to dealing with such problems. Finally, if patients prefer an intervention that is more costly than the status quo, logic dictates that those extra resources will have to be obtained from another health-care programme. In such contexts, to aid decision making, values derived from members of the community for different programmes may be more relevant than values derived from patients. Initial studies in the use of WTP in this broader context of eliciting community values are also outlined. CONCLUSIONS: WTP has potential, but its application, and interpretation, are not straightforward. More testing of the 'marginal approach' is required and greater use of qualitative research, to assess the validity of the approach, should be made in this area. PMID- 11493322 TI - Giving tape recordings or written summaries of consultations to people with cancer: a systematic review. AB - OBJECTIVE: To examine the effects of providing recordings or summaries of consultations to people with cancer and their families. DESIGN: Systematic review. DATA SOURCES: MEDLINE, CINAHL, Cancerlit, EMBASE and other electronic bibliographic databases. Bibliographies of relevant papers. SELECTION CRITERIA: Randomized and non-randomized controlled trials of the provision of taped recordings or written summaries of consultations to people with cancer and/or their families. MAIN RESULTS: Eight randomized controlled trials were found, all involving adult participants. No non-randomized controlled trials were found. The quality of the studies was generally poor. Between 83% and 96% of people who received recordings or summaries found them useful to remind them of what was said and/or to inform family members and friends about their illness and treatment. Of seven studies that assessed recall of information given during the consultation, four reported better recall among the groups that received recordings or summaries than among control groups. Receiving a recording or summary had no significant effect on anxiety or depression between the groups. None of the included studies assessed survival or health outcomes other than psychological outcomes. CONCLUSIONS: Wider use of consultation tapes and summary letters could benefit many adults with cancer, without causing additional anxiety or depression, but consideration should be given to individuals' circumstances and preferences. PMID- 11493325 TI - Shaping the trajectory of patients with venous ulceration in primary care. AB - OBJECTIVE: To explore the patient's experience of venous ulceration and how it is shaped within primary care. DESIGN: Qualitative grounded theory study. PARTICIPANTS AND SETTING: Thirty-nine patients, 33 nurses and 14 general practitioners in a major health district in England. RESULTS: The findings indicate that patients with the chronic condition of venous hypertension are handled in an anomalous way within primary care when they present with ulcers on their lower limbs. The trajectory projections for the patients are not developed from the usual basis of a medically defined condition-specific diagnosis but from a symptom-specific diagnosis. This leads to an unusual context of care where there is a serious but unrecognized conflict of focus between the nurses and their patients. The nurses in this study tended to set priorities related to the ulcer and to the underlying pathology, whereas the patients wanted help in pain management and in normalizing their lives. The patients eventually came to a position of 'guarded alliance'2 that in this study took one of three forms: adapting and enduring, emphasizing the positive, or negotiating for comfort. CONCLUSION: The acute care approach applied to patients with this chronic condition led to a situation where the professionals usurped the self-care potential of the patients and navigated rather than piloted them through an acute phase of an underlying chronic illness. This in turn, led to poor quality of life for many of the patients and to frustration for the nurses who were failing to achieve the outcomes they desired. Both perspectives need to be encapsulated into the treatment approach for patients with this condition: healing the ulcer and normalizing the patients' lives can and should form the basis of care. PMID- 11493326 TI - Hepatocellular carcinoma: an overview. AB - Hepatocellular carcinoma remains widely prevalent in tropical Africa and south east Asia and is largely related to chronic hepatitis B virus (HBV) infection. Primary prevention by vaccination of infants at or near birth is effective but any reduction in tumour incidence cannot be expected for decades to come yet, even in those countries in which the necessary resources exist, as millions of adults remain chronically infected. Meanwhile, the incidence is rising in Japan, Mediterranean countries of Europe, Middle East and North Africa and in the USA, largely due to chronic hepatitis C virus (HCV) infection introduced by the indiscriminate use of unscreened blood and blood products in the recent past. Much has been learned from molecular biological studies on hepatocarcinogenesis incriminating the HBX gene of HBV, the core protein of HCV and a unique guanine to thymine transversion at codon 249 has been observed in cases due to aflatoxin exposure. The subject of precancerous lesions, notably adenomatous/dysplastic nodules and large-cell/small-cell change continues to be a source of much debate and the distinction of nodular lesions in cirrhosis from early carcinoma remains uncertain. Spontaneous regression of hepatocellular carcinoma is rare but it is probably immunologically mediated and treatment by activated T-lymphocytes may reduce recurrence rates after surgery. The positive identification of hepatocellular carcinoma by a liver-specific antibody has greatly facilitated the diagnosis in difficult cases. PMID- 11493327 TI - Liver steatosis in chronic hepatitis C: a morphological sign suggesting infection with HCV genotype 3. AB - AIMS: To identify factors associated with liver steatosis in chronic hepatitis C. METHODS AND RESULTS: Occurrence and severity of liver steatosis in 254 chronic hepatitis C patients were compared with presence of alcohol abuse, body mass index (BMI) >26, history of intravenous drug addiction and hepatitis C virus (HCV) genotype. Steatosis was found in 109 (43%) patients. The occurrence of steatosis was significantly associated with ongoing alcohol abuse (P=0.03) or HCV genotype 3 (P= 0.003), but not with BMI >26. A moderate to severe steatosis was present in 60% of patients infected with HCV genotype 3, irrespective of the presence of alcohol abuse, BMI >26 or history of intravenous drug addiction. Using a multivariable stepwise logistic regression analysis, infection with genotype 3 had an odds ratio (OR) of 10 (95% confidence interval (CI)=4.56-22) for a liver steatosis, whereas the presence of a cirrhosis at histology had an OR=0.256 (95% CI=0.07-0.92). CONCLUSIONS: A moderate to severe degree of steatosis of the liver is a morphological sign suggestive of infection with HCV genotype 3, independent of other risk factors of a fatty liver, but it may disappear at late stages of the disease. PMID- 11493328 TI - Spindle cell squamous carcinoma of the oesophagus: an analysis of 17 cases, with new immunohistochemical evidence for a clonal origin. AB - AIMS: To study the morphological and immunohistochemical characteristics of spindle cell squamous carcinoma of the oesophagus, in order better to understand the histogenesis of this tumour. METHODS AND RESULTS: In this study we analysed the morphological and immunohistochemical characteristics of 17 cases of spindle cell squamous carcinoma of the oesophagus. Most tumours were polypoid, but tumours with an ulcerated and infiltrative pattern were also observed. Histologically, most tumours were of superficial type, with a characteristic morphological aspect consisting of two types of tumour cells, i.e. differentiated squamous cells, and spindle cells with transition zones between the two components. On immunohistochemistry, the squamous cells were positive for cytokeratin and the spindle cells showed variable expression of cytokeratin, vimentin and smooth muscle actin. p53 protein was over-expressed in 10 cases, both tumour cell types showing strong nuclear positivity. In most tumours, E cadherin was expressed in the squamous cells and absent in the spindle cells. CONCLUSIONS: The similar pattern of p53 protein expression in the two tumour cell types of spindle cell squamous carcinoma of the oesophagus suggests their common origin. The change in adhesion molecule expression with loss of E-cadherin expression may be associated with the acquisition of spindle cell morphology by the squamous tumour cells. PMID- 11493329 TI - Cytoskeletal and kinetic epithelial differences between NSAID gastropathy and Helicobacter pylori gastritis: an immunohistochemical determination. AB - AIMS: Distinguishing histological features between non-steroidal anti inflammatory drug (NSAID) gastropathy and Helicobacter pylori gastritis have been accepted. However, the molecular basis explaining these dissimilar histologies has not been elucidated. In an attempt to clarify this question we investigated the differences in the structural cytoskeleton and proliferative activity of these two gastropathies. METHODS AND RESULTS: We assessed the distribution of five cytokeratins (CK) (CK7, 8, 18, 19 and 20) and Ki67 for the ability to distinguish NSAID from H. pylori gastropathies. In H. pylori gastritis, CK7, 8, 18 and 19 were expressed comparably to normal mucosa from the deep foveolae up to the tips of the glands. The detection of CK20, normally expressed in the upper foveolar region and surface, was decreased with only an epithelial surface reaction. In NSAID gastropathy, CK expression was increased in intensity, with normal distribution for CK8, 18 and 19. Modification of localization was noted for CK7 and 20, with labelling extending toward the deep foveolar region. Unlike H. pylori gastritis, no surface epithelial labelling with Ki67 was noted with NSAID gastropathy but downward elongation of the proliferative zone occurred instead. CONCLUSIONS: Contrasting cytostructural alterations and distinct proliferative patterns distinguish NSAID gastropathy from H. pylori gastritis, possibly reflecting different injury pathways. PMID- 11493330 TI - Different patterns of beta-catenin expression in gastric carcinomas: relationship with clinicopathological parameters and prognostic outcome. AB - AIMS: The cadherin-catenin complex is known to play a critical role in maintenance of cell adhesion. Additionally beta-catenin (beta-ct) can also take part in signal transduction and nuclear beta-ct expression could be correlated with poor prognosis in several malignancies. Since, in gastric cancer, this role of beta-ct is still uncertain, we investigated the expression pattern of beta-ct as well as the possible prognostic role. METHODS AND RESULTS: beta-catenin expression was immunohistochemically investigated in a retrospective series of 401 R0-resected gastric carcinomas. Out of these cases, 54 tumours (13.5%) revealed a preserved membranous beta-ct expression similar to that in normal gastric mucosa. In 80 tumours beta-ct expression was moderately reduced and in 117 tumours highly reduced. In 150 tumours (37.4%), no or only a weak membranous beta-ct expression was found. Additionally, in 53 tumours, a strong beta-ct expression could be observed in the cytoplasm with a simultaneous nuclear beta-ct immunoreactivity in 17 of these 53 tumours, while nine tumours only showed nuclear immunoreactivity without cytoplasmic staining. There were no significant correlations between the degree of membranous beta-ct expression or the different staining pattern (membranous vs. cytoplasmic/nuclear) and the grade of tumour differentiation, the histological tumour type according to Lauren, as well as with the prognostic parameters pT, pN category and vascular invasion. No associations could be found with tumour cell proliferation and the expression of E-cadherin, irrespectively of the different beta-ct staining pattern. Univariate analysis revealed no influence on survival, either for membranous or for cytoplasmic/nuclear beta-ct expression. CONCLUSION: Our data on 401 tumours suggest that activation of the Wnt/beta-catenin signalling does also occur in a subset of gastric carcinomas. However, in gastric cancer, neither the presence of cytoplasmic/nuclear beta-ct expression nor the reduction or loss of membranous beta-ct expression is correlated with a specific histological tumour type, tumour progression or prognosis. PMID- 11493331 TI - Tryptase-positive mast cells accompany lymphocytic as well as lymphoplasmacytic lymphoma infiltrates in bone marrow trephine biopsies. AB - AIMS: To investigate the specificity of increased bone marrow mast cell numbers in lymphoplasmacytic lymphoma (LPL) and to evaluate the relationship between mast cell number and the immunoglobulin phenotype of neoplastic lymphoid cells. METHODS AND RESULTS: Retrospective study of bone marrow trephine biopsy specimens from patients with LPL, compared with selected cases representing chronic lymphocytic leukaemia (CLL) and multiple myeloma (MM) of known immunoglobulin light and heavy chain phenotype. Bone marrow mast cells were counted following immunohistochemical staining of sections for mast cell tryptase. We have confirmed previous observations that mast cell numbers are increased in bone marrow infiltrates of LPL. However, we found similarly high mast cell numbers in CLL. High mast cell numbers were associated with neoplastic lymphoid cells expressing an IgM kappa phenotype. Mast cell numbers were low in all cases of MM studied and in controls with no lymphoma present. We observed an apparent bias towards kappa light chain expression in our cases of LPL. CONCLUSIONS: Mast cell number should not be considered a reliable factor in the differential diagnosis of LPL and CLL when assessing bone marrow histology. Possible bias towards kappa light chain expression in LPL requires further study, as do the mechanism and functions of mast cell recruitment by neoplastic lymphoid cells. PMID- 11493332 TI - CD10 and Bcl10 expression in diffuse large B-cell lymphoma: CD10 is a marker of improved prognosis. AB - AIMS: Diffuse large B-cell lymphoma (DLBCL), the most common subtype of non Hodgkin's lymphoma, is clinically and pathologically heterogeneous. The Bcl10 gene was recently isolated from the breakpoint region of t(1;14)(p22;q32) in mucosa-associated lymphoid tissue (MALT) lymphomas, and is considered to be an apoptosis-associated gene. CD10 is considered to be a marker of follicular centre B-cell differentiation. To assess the clinical significance and roles of CD10 and Bcl10 in DLBCL, we analysed 138 cases, using immunohistochemical methods. METHODS AND RESULTS: CD10 expression was limited to the cytoplasm, whereas Bcl10 expression was detected in the cytoplasm and/or nuclei. CD10 expression was detected in 39 of 138 cases (28.2%), cytoplasmic Bcl10 in 68 cases (49.2%), and nuclear Bcl10 in 34 cases (24.6%). Nuclear Bcl10 was detected in 14 of 28 cases (50%) of extranodal DLBCL, but only 20 of 110 cases (18.2%) of nodal DLBCL. Cytoplasmic Bcl10 was detected in 19 of 28 cases (67.8%) of extranodal DLBCL and 49 of 110 cases (44.5%) of nodal DLBCL. CD10 expression closely correlated with improved survival (68% overall survival (OS) vs. 48% OS), but not with site of disease. A high International Prognostic Index (IPI) was considered to be a poor prognostic factor associated with a shorter OS. CD10 expression was detected in 27 of 84 cases (32.1%) with low-risk IPIs, and in 12 of 54 cases (22.2%) with high-risk IPIs. In the low-risk group, cases expressing CD10 carried a better prognosis than CD10- cases (93% OS vs. 71% OS), whereas this was not the case in the high-risk group (25% vs. 20%). CONCLUSIONS: Bcl10 expression was associated with extranodal DLBCL, but not with prognosis. CD10 expression was closely associated with improved survival, but not with risk as predicted by IPI. Overall, our results suggest that CD10 expression may be useful, in combination with clinical parameters, for determining the prognosis of DLBCL. PMID- 11493333 TI - Evidence for local immunosuppression and demonstration of c-myc amplification in pyothorax-associated lymphoma. AB - AIMS: Pyothorax-associated lymphoma (PAL) develops in the pleural cavity of patients with a long history of pyothorax. Epstein-Barr virus (EBV) is also involved in PAL, similar to lymphomas in immunodeficient patients. Here we examined T-lymphocyte subsets as well as c-myc and REL gene amplification in PAL tissues. METHODS AND RESULTS: We determined the number and distribution of CD4+ and CD8+ T-lymphocytes, to evaluate T-cells in the host immune reaction in seven cases of PAL. As controls, we also studied 10 cases of extranodal diffuse large B cell lymphoma (DLBL) and 10 cases of nodal DLBL. Chromosomal imbalances in PAL were determined by using comparative genomic hybridization (CGH) analysis. The mean numbers of CD4+ and CD8+ and their ratio were significantly lower in PAL than in nodal DLBL. CGH analysis of PAL showed amplification of the 8q24 chromosomal region. In addition, c-myc amplification was found in four cases of PAL by Southern blot analysis. CONCLUSIONS: Our results suggest that the development of PAL may involve a local immunosuppressive environment and that amplification of c-myc might promote tumour progression, as has been described in the development of Burkitt's lymphoma. PMID- 11493334 TI - Expression of nitric oxide synthase isoforms and nitrotyrosine immunoreactivity by B-cell non-Hodgkin's lymphomas and multiple myeloma. AB - AIMS: Nitric oxide synthases (NOS) are isoenzymes that catalyse the synthesis of nitric oxide (NO). The three main NOS isoforms are: NOS1 or neuronal, NOS2 or inducible, and NOS3 or endothelial. NO plays both physiological and pathological roles, depending on its rate of synthesis and concentration, cellular source and microenvironment. Apoptosis is an important biological factor in low-grade lymphomas, and NO is able to prevent apoptosis. In-situ expression of NOS and synthesis of NO have been shown in several malignant tumours, but not in lymphoid neoplasms. This study evaluates whether human B-cell neoplasms express NOS isoforms, and nitrotyrosine (NY), which is usually interpreted as a marker of NO. METHODS AND RESULTS: We studied the expression of NOS-IR isoforms and NY-IR in 16 cases of B-cell non-Hodgkin's lymphoma (NHL) (five follicle centre cell lymphoma, four small lymphocytic/CLL, and seven diffuse large cell lymphoma), and 10 cases of multiple myeloma (MM). NOS1 was expressed in 5/10 cases of MM, and 15/16 cases of NHL. NOS2 was detected in all cases of MM, and in 14/16 cases of NHL, whereas NOS3 was positive in 3/10 of MM and in only in 1/16 cases of NHL. The expression of NY-IR was observed in 70% of MM cases, and in all cases of B-cell NHL, in a dot-like pattern in few tumour cells. CONCLUSIONS: B-cell neoplasms express neuronal and inducible NOS, and nitrotyrosine. Taken together, our results suggest that B-cell neoplasms can produce NO. The role of NO in the biology, diagnosis and prognosis of B-cell neoplasms remains to be established. PMID- 11493335 TI - Endothelial nitric oxide synthase is strongly expressed in malignant mesothelioma but does not associate with vascular density or the expression of VEGF, FLK1 or FLT1. AB - AIMS: To investigate endothelial nitric oxide synthase (eNOS) expression in malignant mesothelioma and its association with expression of vascular endothelial growth factor (VEGF), its receptors FLK1 and FLT1, and vascular density. METHODS AND RESULTS: eNOS, VEGF, FLK1 and FLT1 were studied in 36 histological mesothelioma samples by immunohistochemistry. Two mesothelioma (M14K, M38K) and one non-neoplastic mesothelial cell line (MET-5A) were studied for eNOS mRNA expression by reverse transcriptase-polymerase chain reaction (RT PCR). Vascular density was determined by staining the samples with an antibody to factor VIII. RT-PCR showed that mesothelioma cells synthesize eNOS in vitro. eNOS immunoreactivity was found in 32/36 (89%) tumours. VEGF, FLK1 and FLT1 expression was found in 17 (45%), 24 (69%) and 25 (71%) cases, respectively. FLK1 or FLT1 immunoreactivity was more often seen in epithelioid and biphasic mesotheliomas than in sarcomatoid ones (P=0.007 and P=0.011, respectively). There was a significant association between FLK1 and FLT1 immunoreactivity (P=0.032). No significant association was found between FLK1, FLT1, VEGF and eNOS immunoreactivity and vascular density. CONCLUSIONS: eNOS is strongly expressed in malignant mesothelioma. Since eNOS did not associate with VEGF, FLK1 or FLT1, its synthesis seems not to be regulated through VEGF in malignant mesothelioma as has been shown in non-neoplastic endothelial cells. PMID- 11493336 TI - Malignant peripheral nerve sheath tumours: high Ki67 labelling index is the significant prognostic indicator. AB - AIMS: We investigated p53, Ki67, MDM2, and p21WAF1/CIP1 in order to evaluate its relationship with prognosis in malignant peripheral nerve sheath tumours (MPNST). METHODS AND RESULTS: In 49 cases of MPNSTs, the immunohistochemical studies of Ki67, p53, MDM2, p21WAF1/CIP1 and polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) with direct sequencing of p53 were performed with the formalin-fixed paraffin-embedded tissue. In 43 cases with survival data available, an evaluation of the prognostic significance of clinicopathological factors was also carried out. A high Ki67 labelling index (LI) (>25%) was correlated with a reduced survival rate in the 43 cases of MPNST (P=0.0106, log rank test). Furthermore, there was a significant correlation between the Ki67 LI and the immunohistochemical expression of p53 or MDM2. In 17 MPNST cases, PCR amplification of exons 5 through 8 of the p53 gene was successful. One case showed a base change of codon 240 (AGT-->AGC), but translated amino acid (Ser) remained unchanged. Multivariate Cox analysis of our series showed that the association of von Recklinghausen's disease, tumour depth, and the presence of rhabdomyoblasts (malignant triton tumour) each had an independent negative impact on overall survival. CONCLUSION: High Ki67 LI (>25%) was of significant prognostic value in MPNST. PMID- 11493337 TI - Immunohistochemical localization of gross cystic disease fluid protein-15, -24 and -44 in ductal carcinoma in situ of the breast: relationship to the degree of differentiation. AB - AIMS: Three major proteins present in breast gross cystic disease fluid and expressed by the cyst lining apocrine epithelium are gross cystic disease fluid protein-15 (GCDFP-15), apolipoprotein-D (APO-D; GCDFP-24) and zinc alpha2 glycoprotein (ZnGP; GCDFP-44). The aim of this study was to investigate the expression of these proteins in ductal carcinoma in situ (DCIS) of the breast and to relate their expression with the degree of differentiation of DCIS. METHODS AND RESULTS: An immunohistochemical study of these proteins was performed in 57 cases of DCIS and nine cases of morphologically apocrine DCIS. Positivity was seen in 24/57 (42.1%) cases with anti-GCDFP-15, 20/57 (35.1%) cases with anti GCDFP-24 and 22/57 (38.6%) cases with anti-GCDFP-44. GCDFP-15 positivity was noted in 5/13 (38.5%) of the well-differentiated, 11/19 (57.9%) intermediately differentiated and 8/25 (32.0%) of the poorly differentiated cases (P=0.217). GCDFP-24 positivity was seen in 3/13 (23.0%) well-differentiated, 9/19 (47.4%) intermediately differentiated and 8/25 (32.0%) poorly differentiated cases (P=0.336). GCDFP-44 was detected in 5/13 (38.5%) of well-differentiated cases, 11/19 (57.9%) intermediately differentiated and 6/25 (24.0%) poorly differentiated cases (P=0.074). In the nine cases of apocrine DCIS, GCDFP-15 positivity was detected in seven (77.8%), while five (55.6%) and six (66.7%) cases were positive for GCDFP-24 and GCDFP-44, respectively. CONCLUSIONS: The results indicate that there is no significant association between the expression of the studied proteins and the degree of differentiation of DCIS of the breast. Moreover, some morphologically apocrine DCIS cases appear to lose expression of these proteins. PMID- 11493340 TI - Dermatofibroma (fibrous histiocytoma): an inflammatory or neoplastic disorder? PMID- 11493338 TI - mRNA expression of urokinase and plasminogen activator inhibitor-1 in human crescentic glomerulonephritis. AB - AIMS: Weak staining for urokinase-plasminogen activator (uPA), tissue type plasminogen activator (tPA), or plasminogen activator inhibitor-1 (PAI-1) confined to crescents has been described in a few cases of severe crescentic glomerulonephritis. We evaluated the molecular mechanism by which these proteins are increased or induced within crescents. METHODS AND RESULTS: We examined uPA, tPA and PAI-1 mRNA expression in 12 renal biopsies with crescentic glomerulonephritis, and in six control renal biopsies with no detectable abnormalities by RNA in-situ hybridization. The expressions of uPA, tPA and PAI-1 proteins were also assessed by immunofluorescence. To better determine the cellular origin of uPA and PAI-1 transcripts, CD68 protein was studied by immunohistochemistry on the same sections on which in-situ hybridization had been performed. In controls, there were very low level signals of uPA and PAI-1 mRNAs in a few glomerular epithelial cells (GECs). Specific signals of uPA and PAI-1 mRNAs were detected in the cells forming crescents in all the cases with crescentic glomerulonephritis. However, weak expression of mRNA for tPA was detected in two cases only. Immunostaining for uPA and PAI-1 was positive in some but not all, cases of crescentic glomerulonephritis. A double-labelling study showed that the signal for PAI-1 and uPA mRNAs was mainly in CD68- cells. CONCLUSIONS: Local accumulation of uPA or PAI-1 in crescents is associated with enhanced mRNA expression of these proteins. The up-regulation of PAI-1 mRNA by GECs, in particular, could play a major role in the formation of persistent fibrin deposits and progression of the lesions in crescents. Whether up regulation of uPA is an epiphenomenon or plays a pathogenic role in the formation of crescents remains to be clarified. PMID- 11493341 TI - Angiomyolipoma of the liver. PMID- 11493342 TI - Prominent cellular whorls within an epithelioid histiocytoma. PMID- 11493343 TI - Spindle cell lipoma with extramedullary haematopoiesis. PMID- 11493344 TI - Myofibrosarcoma of the nasal cavity and paranasal sinus. PMID- 11493345 TI - Systemic and local effects of long-term exposure to alkaline drinking water in rats. AB - Alkaline conditions in the oral cavity may be caused by a variety of stimuli, including tobacco products, antacids, alkaline drinking water or bicarbonate toothpaste. The effects of alkaline pH on oral mucosa have not been systematically studied. To assess the systemic (organ) and local (oral mucosal) effects of alkalinity, drinking water supplemented with Ca(OH)2 or NaOH, with pH 11.2 or 12 was administered to rats (n = 36) for 52 weeks. Tissues were subjected to histopathological examination; oral mucosal biopsy samples were also subjected to immunohistochemical (IHC) analyses for pankeratin, CK19, CK5, CK4, PCNA, ICAM 1, CD44, CD68, S-100, HSP 60, HSP70, and HSP90. At completion of the study, animals in the study groups had lower body weights (up to 29% less) than controls despite equal food and water intake, suggesting a systemic response to the alkaline treatment. The lowest body weight was found in rats exposed to water with the highest pH value and starting the experiment when young (6 weeks). No histological changes attributable to alkaline exposure occurred in the oral mucosa or other tissues studied. Alkaline exposure did not affect cell proliferation in the oral epithelium, as shown by the equal expression of PCNA in groups. The up-regulation of HSP70 protein expression in the oral mucosa of rats exposed to alkaline water, especially Ca(OH)2 treated rats, may indicate a protective response. Intercellular adhesion molecule-1 (ICAM-1) positivity was lost in 6/12 rats treated with Ca(OH)2 with pH 11.2, and loss of CD44 expression was seen in 3/6 rats in both study groups exposed to alkaline water with pH 12. The results suggest that the oral mucosa in rats is resistant to the effects of highly alkaline drinking water. However, high alkalinity may have some unknown systemic effects leading to growth retardation, the cause of which remains to be determined. PMID- 11493346 TI - Apoptosis of rat gastric mucosa and of primary cultures of gastric epithelial cells by indomethacin: role of inducible nitric oxide synthase and interleukin-8. AB - In order to gain insights into indomethacin-induced gastric injury, rats were fed with indomethacin (20 mg/kg), or alternatively, the primary cultures of rat gastric epithelial cells were cultured with different doses of indomethacin (1 1000 microM). Light microscopy, electron microscopy, fluorescence microscopy, TdT mediated dUTP-biotin nick end labelling staining, ssDNA staining and DNA fragmentation assay were employed to evaluate the levels of gastric injury and apoptosis. Cells expressing inducible nitric oxide synthase (iNOS) and interleukin (IL)-8 were localized at the rat gastric mucosa by immunohistochemistry. Administration of indomethacin to rats caused apoptosis and injury of the gastric mucosal epithelial cells. Indomethacin also induced apoptosis of primary cultures of gastric epithelial cells in a dose-dependent manner. Cells expressing iNOS and IL-8 were detected at and around the sites of gastric injury in the indomethacin-fed rats, but not in the control rats. The induction of apoptosis by indomethacin in the primary cultures of gastric epithelial cells suggests that the direct apoptotic capacity of indomethacin. iNOS and IL-8 may be involved in this process. PMID- 11493347 TI - Peplomycin, a bleomycin derivative, induces myofibroblasts in pulmonary fibrosis. AB - To analyse the mechanism by which a bleomycin derivative, peplomycin (PLM) induces pulmonary fibrosis, we investigated differentiation of rat pulmonary fibroblasts to myofibroblasts (MF). In intraperitoneally PLM (5 mg/kg/day) injected rats, the peripheries of lungs adjacent to the pleura revealed advanced fibrosis with a small number of alpha-smooth muscle actin (alpha-SMA)-positive MF, which ultrastructurally possessed abundant microfilaments and cellular organelles. In the fibrotic tissue, the expression of alpha-SMA-mRNA was detected by in situ reverse transcription-polymerase (RT-PCR). The message was strong just after a 2-week administration of PLM then decreased thereafter, although fibrosis advanced. When pulmonary fibroblasts were separated from saline-injected rats (N Fib) and cultivated for 7 days in the presence of 5 mg/mL PLM, alpha-SMA protein was weakly expressed, while the majority of pulmonary fibroblasts separated from PLM-injected rats (P-Fib) became positive for alpha-SMA in 7-day cultivation and the expression of alpha-SMA in P-Fib was strongly increased by cultivation in the presence of PLM and transforming growth factor-beta (TGF-beta), but not basic fibroblast growth factor (bFGF) or platelet-derived growth factor (PDGF), although the cell proliferation was most strongly enhanced by bFGF and only slightly by PLM and TGF-beta. The alpha-SMA-positive cells expressed vimentin, but only weakly expressed desmin. Additionally, P-Fib generated larger amounts of TGF-beta and bFGF than were generated by N-Fib. These results indicate that PLM induces pulmonary fibrosis by differentiating fibroblasts to alpha-SMA-positive MF, and that bFGF and TGF-beta play each critical role in the different phases of PLM-induced pulmonary fibrosis by inducing fibroblast proliferation and transformation, respectively. PMID- 11493348 TI - Changes in serum concentration of antioxidants following treadmill exercise testing in patients with suspected ischaemic heart disease. AB - Twenty-four subjects with suspected ischaemic heart disease underwent a treadmill exercise stress test (TEST). Nine individuals developed ischaemia as defined by standard criteria. Total plasma antioxidant status (TPAS), and serum concentrations of vitamin E were measured pre-TEST, and 0, 1, 2, 4, 8 and 24 h following the treadmill test. Mean serum vitamin E concentrations fell by 33% in the group as a whole (from 9.53 +/- 0.92 mg/L pre-TEST to 6.39 +/- 1.06 mg/L immediately post stress test, P < 0.02) and rose to baseline over the subsequent 24 h. The levels of serum vitamin E fell by 34% in the group of patients who had a positive TEST, and 32% in those who did not develop ischaemia during the TEST. Serum cholesterol concentrations also fell significantly during the TEST. In the total group serum cholesterol fell by 6.5% (P = 0.0052), and in the subgroup who were positive for ischaemia the fall in serum cholesterol was 10.3% (P = 0.004). The reduction in serum cholesterol was 4.1% in the subgroup who did not develop ischaemia (P > 0.05). Mean total plasma antioxidant status showed no significant temporal change for the group as a whole, although there was a nonsignificant decrease immediately post-TEST in the ischaemic group and a slight rise at 8 h in the group negative for ischaemia. PMID- 11493349 TI - Association between oral malodor and adult periodontitis: a review. AB - BACKGROUND: Bad breath has a significant impact on our daily social life to those who suffer from it. The majority of bad breath originates within the oral cavity. However, it is also possible that it can come from other sources such as gastric intestine imbalance. The term "oral malodor" is used to describe a foul or offensive odor emanating from the oral cavity, in which proteolysis, metabolic products of the desquamating cell, and bacterial putrefaction are involved. Recent evidence has demonstrated a link between oral malodor and adult periodontitis. The process of developing bad breath is similar to that noted in the progression of gingivitis/periodontitis. Oral malodor is mainly attributed to volatile sulfur compounds (VSC) such as hydrogen sulfide, methyl mercaptan and dimethyl sulfide. The primary causative microbes are gram-negative, anaerobic bacteria that are similar to the bacteria causing periodontitis. These bacteria produce the VSC by metabolizing different cells/tissues (i.e., epithelial cells, leukocytes, etc.) located in saliva, dental plaque, and gingival crevicular fluid. Tongue surface is composed of blood components, nutrients, large amounts of desquamated epithelial cells and bacteria, suggesting that it has the proteolytic and putrefactive capacity to produce VSC. One of the challenges in dealing with oral malodor is to identify a reliable test for detecting bad breath. AIMS: The purposes of this review article were: (1) to correlate the relationship between oral malodor and adult periodontitis; (2) to analyze current malodor tests and discuss available treatment regimens. PMID- 11493350 TI - Subgingival debridement of root surfaces with a micro-brush: macroscopic and ultrastructural assessment. AB - AIM: The aim of this study was to assess the use of a micro-brush to remove plaque deposits from subgingival, periodontally involved root surfaces in vivo. METHODS: 30 periodontally involved teeth requiring extraction for periodontal or prosthetic reasons in 26 adult patients were utilised. For inclusion, teeth had to display at least 30% bone loss radiographically. Following the establishment of local anaesthesia, grooves were cut on the proximal root surface adjacent to the gingival margin at the line angles. For each tooth, 1 proximal root surface was rubbed with the micro-brush for 2 min to the depth of the pocket whilst the other root surface acted as an undebrided control. The teeth were then extracted, rinsed in 0.85% NaCl, stained with 2% erythrosine solution and photographed. The amount of erythrosine staining on each subgingival, periodontally involved root surface was assessed by tracing the areas of stain on a colour photograph and scanning the tracings into a computerised image tracing program. RESULTS: Results were expressed as the % of the periodontally involved root-surface area that exhibited staining. Stained areas were further examined with the scanning electron microscope (SEM). The undebrided root surfaces each displayed 100% staining. The debrided surfaces (with probing pocket depths of 4-10 mm) displayed mean staining of 16.1% (SD +/-7.1%) of the proximal surface area. SEM assessment showed that undebrided root surfaces were covered with thick deposits of bacteria. On debrided surfaces, stain-free areas were free of plaque whilst areas of faint staining exhibited either no plaque, calculus deposits or scanty, isolated islands of bacteria. Bacteria had been partially removed from the surface of calculus in some areas. CONCLUSIONS: The findings indicate that subgingival debridement with a micro-brush is effective in removing plaque deposits from periodontally involved root surfaces. PMID- 11493351 TI - Interleukin-10 gene promoter polymorphism in Japanese patients with adult and early-onset periodontitis. AB - BACKGROUND: IL-10 is an anti-inflammatory cytokine, which may modulate disease expression in chronic inflammatory periodontal disease. 3 dimorphic polymorphisms within the IL-10 gene promoter have recently been identified and appear to influence regulation of its expression. AIM: The aim of the present study was to investigate whether the promoter polymorphisms are associated with adult periodontitis (AP) and generalized early-onset periodontitis (G-EOP). METHODS: Genomic DNA was obtained from 34 AP patients, 18 G-EOP patients and 52 controls. The promoter region between -506 and -1140 was amplified by polymerase chain reaction, and polymorphisms were detected by nucleotide sequencing. RESULTS: The haplotype frequencies in Japanese were quite different from those of Caucasian and were even slightly different from those of southern Chinese with systemic lupus erythematosus. We found no significant difference in allele or haplotype frequencies between patients and controls. CONCLUSIONS: IL-10 production may be regulated within the complex cytokine network in chronic inflammatory periodontal disease, rather than the gene polymorphisms. PMID- 11493352 TI - Systemic and local antimicrobial use in periodontal therapy in England and Wales. AB - BACKGROUND/AIMS: The aim of this study was to investigate antimicrobial use during periodontal therapy in dental practice in England & Wales. METHOD: This was a postal questionnaire survey of 800 dentists, 400 general dental practitioners (GDP) in National Health Service practice and 400 members of the British Society of Periodontology (Periodontal Society) primarily in dental practice. We designed and piloted a questionnaire to evaluate both systemic and local antibiotic use with periodontal therapy as well as factors affecting their prescription. In addition, we also investigated the potential use of antibiotic sensitivity testing, since this has been recommended prior to prescribing antibiotics. Two follow-up mailings were used to encourage non-responders. RESULTS: The useable return rate for the questionnaires was 587/800 (73%). Systemic antibiotics were used by 7.4% Periodontal Society members and 18.4% GDP for untreated adult periodontitis patients (p<0.001). Antimicrobials were prescribed more frequently by Periodontal Society members in early onset (52.7%) and refractory periodontitis patients (49.6%), and this was highly statistically significantly greater usage than GDP (p<0.001). Regarding local antimicrobials, usage for untreated adult periodontitis was Periodontal Society 8.9% and GDP 5.4%. Higher usage of local antimicrobials was found both for the treatment of recurrent pocketing in adult periodontitis (Periodontal Society 26.3%, GDP 14.8%, p<0.014) and refractory periodontitis (Periodontal Society 30.8%, GDP 15.2%, p<0.001). As reasons for using local antimicrobials, more than 80% of all respondents stated superiority over root debridement alone. Barriers to use included cost, no perceived need and lack of supporting research data. The percentage of responders considering diagnostic microbiology either theoretically or at a cost of pound 60 were by group, Periodontal Society 83% & 70.4% and GDP 76% & 51.2%. 33% of Periodontal Society members and 3.8% of GDP spent at least 45 min per quadrant on root planing and Periodontal Society members had a greater exposure to lectures on both systemic and local drug therapy compared with GDP (p<0.001). CONCLUSIONS: Systemic antimicrobial use was infrequent for adult periodontitis and generally in line with current recommendations for other disease types. Whilst local antimicrobial therapy for periodontitis was not widespread, a substantial minority of dentists use this form of therapy and most believe that it is more effective than root debridement alone. PMID- 11493353 TI - Cytokine gene expression in chronic periodontitis. AB - BACKGROUND: Cytokines play an important role in controlling inflammatory processes and tissue homeostasis. Periodontitis, as any other chronic inflammatory disease, results from a disarrangement of host factors, mainly cytokines and the initiating agent. Modulation of the cytokines is not only controlled by the host but also by infecting bacteria and their products. AIM: In the present study, we examined the cytokine mRNA expression profiles in six patients, each presenting sites affected with (1) severe progressive periodontitis, (2) chronic, but stable periodontal lesions, and (3) with healthy sites. Analysis using a quantitative RT-PCR included IFN-gamma, IL-1beta, IL-2, IL-4, IL-5, IL-6, and TNF-alpha. MATERIAL AND METHODS: 6 patients with chronic periodontitis were following treatment observed for a period of six years for local sites staying healthy, local sites with periodontal pathology but without signs of progression of attachment loss and sites with verified progression were biopsied. The biopsies were lyzed and analyzed for levels of cytokine mRNAs. RESULTS: Results revealed considerable variation not only between patients, but also between individual sites. Each patient's site has thus to be looked at as an independent entity. CONCLUSIONS: The local action of cytokines, which is heavily dependent on recruitment, interaction and activation of immunocompetent cells can explain the site-specific nature of cytokine expression. Cytokine data from individual sites together with the local clinical status and data from the literature demonstrate the complexity of periodontal disease pathogenesis. To gain insight to specific mechanisms further studies are needed. PMID- 11493354 TI - Tacrolimus is not associated with gingival overgrowth in renal transplant patients. AB - BACKGROUND: Cyclosporin A is used extensively to prevent the rejection of allogenic renal transplants. However, it is associated with a variety of undesirable side effects including gingival overgrowth. Tacrolimus (FK506), has been marketed as an effective alternative immunosuppressant to cyclosporin A and recent subjective reports suggest patients taking it complain infrequently of gingival problems. This clinical investigation was undertaken to confirm whether or not tacrolimus adversely affected the gingival health of renal transplant recipients. METHODS: Renal transplant patients (RTPs) under the care of the Renal Transplantation Service at the Manchester Royal Infirmary, who had received a renal allograft at least 18 months earlier, were recruited for this study. All but one of the RTPs had been taking tacrolimus since transplantation. The other had commenced tacrolimus therapy two months after receiving her allograft. A hospital based control group was recruited from non transplanted individuals attending the Turner Dental School, Manchester. Each patient underwent a detailed dental assessment and had dental impressions taken. The extent of gingival overgrowth was determined from plaster models. RESULTS: 25 renal transplant recipients and 26 control patients were included in the study. None of the individuals in either the tacrolimus or control groups had clinically significant overgrowth. The patients in the tacrolimus group with the highest overgrowth scores were those also taking calcium antagonists as treatment for hypertension. CONCLUSION: This study demonstrates that tacrolimus has no adverse effects on the gingival tissues and thus has potential as an alternative immunosuppressant for individuals susceptible to developing cyclosporin A-induced gingival overgrowth. PMID- 11493355 TI - Porphyromonas gingivalis-specific IgG subclass antibody levels as immunological risk indicators of periodontal bone loss. AB - BACKGROUND: It has been well demonstrated a positive association between the magnitude of host antibody response and periodontal disease status. Previous studies also reported that Porphyromonas gingivalis-specific IgG subclass antibodies were elevated in sera from adult periodontitis patients. However, the role and the association of these IgG subclass antibodies to the development of periodontal diseases are poorly understood. AIM: The aim of present investigation was to examine the relation of serum IgG subclass antibody levels and alveolar bone loss in treated and untreated periodontitis patients. METHODS: Serum samples were taken from 20 treated and maintained periodontitis patients (SPT patients), 30 untreated patients and 19 periodontally healthy subjects. We determined the IgG subclass antibody titers to P. gingivalis whole cells using an enzyme-linked immunosorbent assay (ELISA). Subgingival plaque samples were taken from the mesio buccal surface of 6 randomly selected teeth of SPT patients and evaluated for the presence of P. gingivalis by immunofluorescence microscopy. Clinical measurements were also taken including full mouth intraoral radiographs to measure interproximal alveolar bone loss at baseline (BLS1) and at a 5-year recall visit in the SPT patients (BLS2). RESULTS: Our results indicated that both patient groups had detectable levels of IgG1, IgG2, and IgG4. Significantly higher IgG1 was observed in both patient groups compared to the healthy subjects. The untreated patients also exhibited significantly elevated IgG2 response (p<0.05). The mean IgG4 level of the SPT patients was significantly higher compared to the other subject group (p<0.05). A statistically significant positive correlation between IgG2 levels and changes in bone levels (DeltaBLS: BLS2-BLS1) was seen in the SPT patients (p<0.001). SPT patients with high IgG2 and low IgG4 showed greater bone loss than those with low IgG2 and high IgG4 (p<0.05), although the mean prevalence of P. gingivalis in the 2 groups did not differ significantly. CONCLUSION: Our data suggest that the prolonged IgG2 response after periodontal treatment may be indicative of recurrent or persistent periodontal destruction. PMID- 11493356 TI - Yeasts in periodontal pockets. AB - OBJECTIVES: The presence of yeasts in periodontal pockets has been described in a few studies. The association between yeasts and putative periodontal pathogens is not well described. This study aims at assessing the prevalence of yeasts in periodontal pockets and possible associations with the clinical conditions of the sampled sites and other micro-organisms present. MATERIAL AND METHODS: 2 subject groups form the basis for this study. The 1st comprises results from microbiological samples from periodontal pockets of 128 subjects. The 2nd originates from 126 periodontal patients with untreated pockets. Microbiological identification was performed after cultivation on blood and Sabouraud agar plates, and "checkerboard" DNA-DNA hybridisation. RESULTS: The prevalence of subjects with yeasts in the pockets was 15.6% and 17.5% in the 2 groups respectively and was inconsistent according to gender. No correlation was found between age and the presence of yeasts. Eubacterium saburreum was weakly correlated with presence of yeasts (r=0.194 p=0.03). Yeasts were rarely found in both samples from the same individual. CONCLUSIONS: Our results indicate that yeasts can be expected to be present in periodontal pockets in one out of 6 periodontal patients independent of gender and age. Eubacterium saburreum seems to occur frequently together with yeasts. PMID- 11493357 TI - On the association between hypercholesterolemia, cardiovascular disease and severe periodontal disease. AB - BACKGROUND: Premature death in men is known to be significantly associated with coronary heart disease (CHD). More and more studies are pointing toward a possible association between periodontal disease and increased risk of cardiovascular disease. The association of poor oral hygiene and atherosclerosis can be explained by the effect of chronic inflammatory disease on blood rheology. The purpose of the present study was to assess the relationship between CHD and periodontal disease. PATIENTS AND METHODS: The study population included 1094 Israeli army service men aged 26-53 years (mean: 39+/-5 years). The study group comprised 151 subjects classified as having coronary heart disease CHD, i.e., myocardial infarction, and or anginal syndrome with angiographic evidence of significant coronary disease, or suffer from atherosclerotic risk factors, i.e., diabetes (fasting glucose) and HTN according to strict, well-established criteria. Blood levels of cholesterol and triglycerides were also determined. The severity of periodontal disease was assessed by the aid of CPITN. The control group comprised 943 healthy subjects. Statistical analysis was performed with chi2 test. RESULTS: Statistical analysis showed a significant association of CPITN score 4 with hypercholesterolemia and a possible association with CHD. CONCLUSIONS: The generation of higher cholesterol blood levels is proposed as a possible link between chronic periodontal inflammation and atherosclerosis. PMID- 11493358 TI - Reliability of assessing interproximal bone loss by digital radiography: intrabony defects. AB - AIM: The aim of the present study was to assess the reproducibility and validity of linear measurements of interproximal bone loss in intrabony defects on digitized radiographic images after application of different filters and magnifications. METHODS: Immediately before surgery 50 radiographs of 50 periodontally diseased teeth exhibiting interproximal intrabony defects were obtained by a standardized technique in 50 patients. Intrasurgically the distances from the cementoenamel-junction (CEJ) to the alveolar crest (AC) and from the CEJ to the deepest extension of the bony defect (BD) were assessed. All radiographs were digitized by a flatbed scanner (resolution: 600x1200 dpi). Using the FRIACOM-soft ware, the linear distances CEJ to AC and CEJ to BD were measured at 50 intrabony defects on digitized but unchanged radiographic images and also after use of 2 different basic image processing modes (filters: enhancement of grey level differences, spreading of grey values) with 7-fold and 14-fold magnification by 2 different examiners. RESULTS: Repeated measures MANOVA revealed reproducibility of the measurement of the distance CEJ to AC to be significantly influenced by examiner (p=0.027) and filter in combination with the height of 2 wall component of the intrabony defect (p=0.066). For the distance CEJ to BD filters had significant influence on reproducibility in correlation with vertical angulation difference (p=0.001). On the average in this study radiographic measurements tended to overestimate the amount of bone loss as assessed by intrasurgical measurements (CEJ-AC: 0.74-1.91 mm; CEJ-DB: -0.04-0.77 mm). Validity of measurement of the distance CEJ-AC was shown to be significantly influenced by the depth of the intrabony defect (p<0.003). Validity of the distance CEJ-BD was significantly influenced by intrasurgically assessed bone loss (p=0.029), horizontal angulation (p=0.066). Filters influenced the validity only in combination with examiner (p<0.001). CONCLUSIONS: In this study, the chosen digital manipulations (filters: spreading, structure) of radiographic images failed to result in statistically significantly more reproducible or valid measurements of interproximal bone loss within intrabony defects when compared to the digitized but unchanged images. All radiographic assessments on the digitized images except for use of enhancement of grey level differences (structure) came close to the intrasurgical gold standard. PMID- 11493359 TI - Herpes viruses and periodontopathic bacteria in early-onset periodontitis. AB - OBJECTIVES: This study examined the occurrence of human herpes viruses and suspected periodontopathic bacteria in early-onset periodontitis patients who experienced progressive disease in at least 2 periodontal sites during the maintenance phase of therapy. MATERIAL AND METHODS: In each of 16 individuals (9 male and 7 female; mean age 33.1+/-2.6 years), subgingival plaque samples were collected from 2 deteriorating and 2 stable periodontitis sites. A nested polymerase chain reaction method determined the presence of human cytomegalovirus (HCMV), Epstein-Barr virus type 1 (EBV-1) and herpes simplex virus (HSV). A 16s rRNA polymerase chain reaction method identified Porphyromonas gingivalis, Dialister pneumosintes, Bacteroides forsythus and Actinobacillus actinomycetemcomitans. RESULTS: HCMV was detected in 59.4% of active and in 12.5% of stable sites (p<0.001), EBV-1 in 43.8% of active and in 12.5 % of stable sites (p=0.01), HSV in 34.5% of active and in 9.4% of stable sites (p=0.03), and co infection with any of the 3 test herpesviruses in 43.8% of active and in 3.1% of stable sites (p<0.001). P. gingivalis was detected in 71.9% of active and in 37.5% of stable sites (p=0.01), D. pneumosintes in 62.5% of active and in 18.8% of stable sites (p=0.04), co-infection with P. gingivalis and D. pneumosintes in 50% of active and in 0% of stable sites (p<0.001), and co-infection with any 3 or 4 of the test bacteria in 40.6% of active and in 0% of stable sites (p=0.001). All periodontitis sites showing herpesvirus co-infection and all but one site showing P. gingivalis and D. pneumosintes co-infection revealed bleeding upon probing. CONCLUSIONS: HCMV, EBV-1, HSV and herpesvirus co-infection, as well as P. gingivalis, D. pneumosintes and P. gingivalis-D. pneumosintes co-infection were statistically associated with active periodontitis. Herpesviruses are immunosuppressive and may set the stage for overgrowth of subgingival P. gingivalis, D. pneumosintes and other periodontopathic bacteria. Understanding the significance of herpesviruses in human periodontitis may allow for improved diagnosis, more specific therapy and, ultimately, disease prevention. PMID- 11493360 TI - Prevalence of Actinobacillus actinomycetemcomitans in an ethnic adult Chinese population. AB - AIM: The aim of this study was to determine the prevalence and the structure of the leukotoxin promoter region of Actinobacillus actinomycetemcomitans in an ethnic Chinese population. METHOD: Subgingival plaque samples were collected from 42 patients with moderate to advanced periodontitis and 50 periodontally healthy patients. A. actinomycetemcomitans was detected directly from the crude subgingival plaque by PCR using leukotoxin gene specific primers. The presence of A. actinomycetemcomitans was determined by a single 285 bp PCR amplicon. RESULTS: A. actinomycetemcomitans was found to be present in the subgingival plaque of 68 out of a total of 92 patients examined (74%). 29 out of the 42 periodontitis patients tested were carriers of A. actinomycetemcomitans (69%). Among the periodontally healthy patients studied, 39 out of 50 subjects possessed the bacteria (78%). PCR analysis of the promoter region of the ltx operon revealed that none of the 42 moderate to advanced periodontitis patients examined harboured A. actinomycetemcomitans strains with the JP2-like promoter of the ltx operon, known to enhance leukotoxin expression. 2 out of the 27 advanced periodontitis patients clinically diagnosed as suffering from rapidly progressive periodontitis were found to be carriers of the mildly toxic strain of A. actinomycetemcomitans with the characteristic 652-like promoter. CONCLUSIONS: The high prevalence of A. actinomycetemcomitans, regardless of whether the subgingival samples were analysed from patients with healthy or diseased periodontium suggests that this bacterial species is part of the normal oral flora of ethnic Chinese. Our preliminary results also suggested that subjects who harboured the mildly toxic strain of A. actinomycetemcomitans were potentially susceptible to aggressive forms of periodontitis. PMID- 11493361 TI - Multivariate analysis of oral hygiene data from a representative sample. AB - BACKGROUND: This analysis was based on an oral health survey with dental examination and interview by questionnaire. METHOD: The data set comprised a representative random sample of the over 14-year-old residents of the German Federal State of Saxony (n=714), in which a multivariate logistic regression analysis was conducted. The dependent variable was the debris index (DI) score of the oral hygiene index scoring system. The subjects were split into 2 groups by dividing DI scores into tertiles: those with acceptable oral hygiene (1st tertile, DI 0.00< or =1.20) and unacceptable oral hygiene (2nd and 3rd tertile, DI>1.20). The most important explanatory variable was the community periodontal index of treatment need (CPITN). RESULTS: The higher the CPITN score, the lower the probability of acceptable oral hygiene. In relation to the reference category score 0, higher CPITN scores were associated with acceptable oral hygiene, with an odds ratio 0.05 (95% confidence interval (CI)=0.01-0.23) for CPITN score 1, and an odds ratio 0.02 (95% CI: 0.01-0.08) for score 4. Further significant variables were: self-evaluation of tooth condition, gender, and the number of missing teeth. PMID- 11493362 TI - Tobacco smoking with periodontal disease. PMID- 11493364 TI - Prescribing information systems; making sense of primary care data. PMID- 11493363 TI - Prostate cancer and antioxidants. PMID- 11493365 TI - Record-linkage methodology for prescribing research. AB - Record-linkage refers to the linking together of data relating to the same individual from separate source files. In this paper, we discuss ways in which the technique can enhance observational prescribing research in large populations. We draw upon the work of the Medicines Monitoring Unit (MEMO), University of Dundee, to illustrate its contribution to prescribing research. PMID- 11493366 TI - Treatment of aneurysmal subarachnoid haemorrhage in elderly patients. AB - Spontaneous subarachnoid haemorrhage (SAH) due to ruptured saccular aneurysm is the fourth most frequent cerebrovascular disorder following atherosclerosis, embolism, and primary intracerebral haemorrhage (1). SAH is a common and often devastating condition, which is a significant cause of world-wide morbidity and mortality (2). The aim of this article is to review the epidemiology, pathophysiology and current management of SAH. PMID- 11493367 TI - Pharmacology of oral combination analgesics: rational therapy for pain. AB - No single analgesic agent is perfect and no single analgesic can treat all types of pain. Yet each agent has distinct advantages and disadvantages compared to the others. Hence, clinical outcomes might be improved under certain conditions with the use of a combination of analgesics, rather than reliance on a single agent. A combination is most effective when the individual agents act through different analgesic mechanisms and act synergistically. By activating multiple pain inhibitory pathways, combination analgesics can provide more effective pain relief for a broader spectrum of pain, and might also reduce adverse drug reactions. This overview highlights the therapeutic potential of combining analgesic medications with different mechanisms of action, particularly a nonsteroidal anti-inflammatory drug (NSAID) or acetaminophen with an opioid or tramadol. PMID- 11493369 TI - Effects of Paeoniae Radix, a traditional Chinese medicine, on the pharmacokinetics of phenytoin. AB - BACKGROUND: Phenytoin (PHT), one of the most widely prescribed antiepileptic drugs, has been reported to be associated with numerous drug-drug interactions. However, there are far fewer reports about the pharmacokinetic interactions between PHT and traditional Chinese medicines (TCMs). Paeoniae Radix (PR), one of the well-known TCMs, is used as an adjunct in some epileptic patients. OBJECTIVE: In the present work, we studied the influences of PR on the pharmacokinetics of PHT in rats to identify the possible interactions between PR and PHT. METHOD: A single dose of PHT (100 mg/kg) alone or in combination with PR extract (300 mg/kg) was administered by gavage to male SD rats. Serial blood samples of PHT were obtained for up to 24 h post-administration and measured by high-performance liquid-chromatography. The free (unbound) plasma concentrations of PHT were determined by fluorescence polarization immunoassay. The plasma concentrations were used to construct pharmacokinetic profiles by plotting drug concentration time curves. All data were subsequently processed by the computer program WINNONLIN. Statistical comparisons of pharmacokinetic parameters were performed with the unpaired Student t-test. RESULTS: The mean maximum plasma concentration of PHT was attained 2 h after oral administration of PHT alone and 4-6 h after oral administration of PHT in combination with PR. The plasma level of PHT declined with a half-life of 5.38 h after PHT alone and 4.03 h after PHT and PR given together. No statistically significant differences were obtained in most of the pharmacokinetic parameters (Cmax, AUC, t1/2, MRT and CL/F) and protein binding rates of PHT between the two treatments. However, significant differences in Tmax and Vd/F between groups were noted. CONCLUSION: The significant increase in Tmax indicated that simultaneous oral administration of PR delayed the absorption of PHT. The delayed absorption of PHT might lead to its slow onset of clinical effect. There were no significant differences in Cmax, AUC, t1/2, MRT and CL/F of PHT between the two groups, showing that PR could not significantly affect the extent of absorption, metabolism and elimination of PHT. No significant difference in protein binding rate was found, indicating that PR might not significantly alter the protein binding of PHT. While a significant decrease in Vd/F was noted, the mechanism underlying the apparently decreased Vd/F of PHT influenced by PR needs further study. PMID- 11493368 TI - Plasma concentrations of alpha-tocopherol and urate in patients with different types of cancer. AB - OBJECTIVE: To measure plasma concentrations of alpha-tocopherol and urate to determine whether there is any relationship with different types of cancer before treatment. METHOD: Plasma concentrations of those two antioxidants were measured in cancer patients and sex- and age-matched controls. Plasma total cholesterol concentration was measured to derive an alpha-tocopherol/cholesterol ratio. RESULTS: Alpha-tocopherol and urate concentrations as well as alpha tocopherol/cholesterol ratios were significantly lower in the cancer patients as whole, in gastrointestinal cancer and in breast cancer compared to the controls. There was no significant relationship between alpha-tocopherol and urate levels in either the plasma of patients (r=0.048; P=0.653) or controls (r=0.073; P=0.406). Alpha-tocopherol was more positively correlated with cholesterol in the plasma of controls (r=0.539; P < 0.0001) in comparison with the patients (r=0.456; P < 0.0001). There was no significant correlation between urate and cholesterol levels in both patient and control plasma samples. CONCLUSION: The data suggest that although alpha-tocopherol and urate provide important antioxidant defence in cancer, the association between their levels is weak. PMID- 11493370 TI - Lithium reduces maternal child abuse behaviour: a preliminary report. AB - OBJECTIVE: To find out in child abusing mothers whether aggressiveness in general, but which is directed towards their children in particular, could be reduced by giving lithium. METHOD: An open trial of lithium in eight child abusing mothers. Assessment of aggression using Yudofsky's Overt Aggression Scale. RESULTS: There was a statistically significant reduction (P < 0.05) in aggressiveness to the children and to inanimate objects. CONCLUSION: Lithium could be an effective adjunct in the social and psychological treatment of parents who abuse their children. An adequately powered controlled study testing lithium against placebo over a longer period of treatment and observation would be useful. PMID- 11493371 TI - Assessment of weight-based versus standard dosing of heparin in patients with unstable angina. AB - OBJECTIVE: To assess whether a weight-based dosing regimen (80 u/kg + 18 u/kg/h) or a standard-fixed dose regimen (5000 u + 1000 u/h) of heparin is more appropriate in patients with unstable angina (UA). METHOD: A drug use evaluation was conducted. Patient data for all patients weighing less than 100 kg who were in the coronary care unit of a Veterans Administration Hospital and who received heparin for UA (>24 h) over a 7-month period were included. For the first 4 months, patients were given standard heparin dosing and in the final 3 months patients were given heparin based on weight. The proportion of patients achieving activated partial thromboplastin times (aPTTs) that were at least therapeutic during therapy, the time to achieve the aPTT at a level that was at least therapeutic, and the number of patients with aPTTs over the therapeutic range were compared between groups. RESULTS: Patients in the group receiving weight based heparin therapy (n = 23) were significantly more likely to achieve an aPTT that was at least therapeutic than patients receiving standard therapy (n = 42, 100% vs. 76%, respectively, P = 0.011). When all the patients in each group who achieved an aPTT that was at least therapeutic were compared, the weight-based group achieved the levels significantly faster than the standard-fixed dosing group (7.3 +/- 6.1 vs. 22.6 +/- 17.6 h, respectively, P = 0.0003). However, the use of weight-based dosing was associated with a higher incidence of achieving supertherapeutic aPTTs than standard therapy (78.3% vs. 50.0%, respectively, P = 0.049). CONCLUSION: Patients with UA may achieve therapeutic aPTTs faster than those on standard therapy but they also have a higher risk of achieving a supertherapeutic aPTT. PMID- 11493372 TI - Peak expiratory flow rate and symptom self-monitoring of asthma initiated from community pharmacies. AB - OBJECTIVE: To compare the use of patient-performed peak expiratory flow (PEFR) and symptom monitoring as asthma self-management tools initiated from community pharmacies. DESIGN AND SETTING: 110 patients over 6 years of age were recruited from five private-sector community pharmacies. Patients were identified from pharmacist recall as having 'asthma'. Information on the frequency of their asthma symptoms, medication use, level of physical activity, school or work attendance and lung function was obtained using a questionnaire to classify patients as either mild, moderate or severe. Each patient was alternately assigned to either the symptom or PEFR monitoring procedure in the order they were recruited. Patients performing symptom monitoring used a visual analogue scale to assess symptoms, whereas those in the PEFR monitoring group assessed symptoms and used a pocket-size peak flow meter to measure lung function. Both self-monitoring groups were required to adhere to an individualized management plan based on guideline recommendations and to record their monitored data in a diary card for 2 months. Data from the diary cards were reviewed, collated, transcribed and analysed using the Student t and Mann-Whitney tests. OUTCOME MEASURES: The average monthly frequency of appropriate patient responses determined from their adherence to the self-management plan was used to compare the usefulness of symptom and PEFR self-monitoring. In particular, appropriate use of medication and need for medical consultation was compared. RESULTS: 21 symptom and 40 PEFR-assigned patients completed 2 months' monitoring. The average monthly frequency of appropriate responses in patients using PEFR (0.76) was significantly higher than that of patients using symptom monitoring (0.53, P < 0.006). Patients applying symptom monitoring had a higher monthly frequency (0.39) of inappropriate medication use compared to the PEFR group (0.14). Furthermore, the patients' mean daily symptom scores (2.85) were significantly lower than that estimated by the researcher (4.12, P < 0.03). For all three asthma severity groups a higher monthly average of appropriate responses was observed in patients using PEFR monitoring compared to those who used symptom monitoring. CONCLUSION: PEFR self-monitoring proved to be a more useful asthma tool than symptom self-monitoring. Patients applying symptom monitoring tend to underestimate the severity of their condition and use medication inappropriately. Active involvement of community pharmacists in facilitating and reinforcing out patient self-monitoring would help to optimize asthma management. PMID- 11493373 TI - Influence of adjuvant chemotherapy with cyclophosphamide, methotrexate and 5 fluorouracil on plasma melatonin and chosen hormones in breast cancer premenopausal patients. AB - OBJECTIVE: To investigate the effect of chemotherapy on levels of melatonin in patients with breast cancer. BACKGROUND: In light of reports on the possible oncostatic role of melatonin in breast cancer patients, it is essential to know the influence of adjuvant chemotherapy with cyclophosphamide, methotrexate and 5 fluorouracil (CMF) on plasma melatonin concentration as well as on its contributing factors, e.g. current hormonal state of the organism. The combination therapy is one of the oldest, safest and most commonly prescribed adjuvant treatments. METHOD: Twenty-four breast cancer patients on CMF chemotherapy were studied along with a control group of 16 healthy pre-menopausal women. RESULTS: Plasma melatonin concentration (determined by RIA method) in breast cancer patients prior to treatment did not differ significantly from that of healthy women, but it was significantly increased after the initial cycle of CMF, and significantly increased as compared to a group of healthy women. We did not notice any significant interactions between plasma melatonin and growth hormone, prolactin, estradiol, progesterone, cortisol and met-enkephalin concentrations in all studied groups. CONCLUSION: The possible oncostatic action of melatonin warrants further investigation to elucidate whether the induced increase of blood melatonin concentration is essential to successful CMF chemotherapy. PMID- 11493374 TI - Isoprostane 8-epi-PGF2alpha is frequently increased in patients with muscle pain and/or CK-elevation after HMG-Co-enzyme-A-reductase inhibitor therapy. AB - BACKGROUND: Muscle pains with or without CK-elevation are among the most frequently observed side-effects in patients with hyperlipoproteinemia on various statins. The pathophysiological background, however, remains obscure. METHODS: We examined isoprostane 8-epi-PGF2alpha, a marker of in-vivo oxidation injury, in plasma, serum and urine in these patients at baseline, when muscle problems manifested and different time intervals after withdrawing the respective statin. A healthy control group and a group of untreated patients with hyperlipoproteinemia were run as controls. RESULTS: The majority of patients with muscular side-effects show elevated 8-epi-PGF2alpha in plasma and urine, whereas serum values were elevated only to a lesser extent. Stopping statin therapy or successfully changing to another member of this family of compounds resulted in a normalization of the values in all patients. CONCLUSION: These findings indicate a significant involvement of oxidative injury in the muscular side-effects of statins in patients suffering from hyperlipoproteinemia. PMID- 11493375 TI - Does pregnancy alter melanocytic nevi? PMID- 11493376 TI - S100A6 preferentially labels type C nevus cells and nevic corpuscles: additional support for Schwannian differentiation of intradermal nevi. AB - BACKGROUND: Melanocytic nevi typically show a morphologic sequence of maturation from epithelioid "type A" cells to fusiform, Schwann cell-like "type C" cells with dermal descent. Nevi may also produce Wagner-Meissner-like structures (nevic corpuscles). Previous studies have shown that this maturation of intradermal nevi recapitulates intermediate stages in Schwann cell development. In intradermal nevi, we have evaluated the pattern of S100A6 protein, a form of S100 found in Schwann cells. METHODS: Formalin-fixed, paraffin-embedded archival tissues were evaluated by immunohistochemistry using antibodies specific for S100A6 and S100B in 38 intradermal nevi (IDN). Ten neurofibromas (NF), 3 Schwannomas (SCH), 2 palisaded and encapsulated neuromas (PEN), and 2 granular cell tumors (GCT) were included as positive controls since these lesions have large numbers of Schwann cells. RESULTS: Melanocytic nevi demonstrated preferential anti-S100A6 staining of "type C" cells (36/38; 28 strong, 8 weak) and nevic corpuscles (25/38; 19 strong, 6 weak) compared to "type A" cells (17/38; 17 weak) and "type B" cells (17/38; 4 strong, 13 weak). All NF, SCH, and PEN stained strongly with anti S100A6. Both GCT were negative with anti-S100A6 but positive with anti-S100B. CONCLUSIONS: The pattern of S100A6 expression in intradermal nevi further supports the hypothesis that maturation in these lesions recapitulates features of Schwann cell differentiation. The lack of S100A6 expression by both GCT suggests that these lesions have lost this feature of Schwann cells, which may play a role in their peculiar phenotypic appearance. PMID- 11493377 TI - Recurrent melanocytic nevus: a histologic and immunohistochemical evaluation. AB - BACKGROUND: Recurrent melanocytic lesions may histologically resemble malignant melanoma. METHODS: We evaluated the original nevi (ON) and recurrent nevi (RN) of 15 patients by routine histology and immunohistochemistry (IHC), examining expression of S-100 protein, gp100 (with HMB-45), MART-1, tyrosinase, and the Ki 67 proliferation marker. RESULTS: Compared with ON, RN had a dermal scar, a significantly greater number of melanophages, and a greater extent of cellular atypia including prominent nucleoli and larger cell size. Architecturally, RN showed significantly less symmetry than ON; however, the percentage of junctional cohesive nests, the presence of suprabasal spread, and the degree of confluence were similar between ON and RN. Both ON and RN showed a decrease in expression of gp100 and tyrosinase with increasing depth ("maturation gradient") and low proliferative activity in both the junctional (4.6% for ON vs. 4.13% for RN) and the dermal components (0.93% for ON vs. 1.45% for RN). CONCLUSIONS: RN exhibit a dermal scar, a greater number of melanophages, cytologic atypia, and asymmetry than ON, features that may raise concern about the possibility of malignant melanoma. However, the area with the irregular architectural pattern is restricted to the epidermis and dermis immediately above the scar. In addition, IHC helps to distinguish RN from malignant melanoma; specifically, RN demonstrate an immunohistochemical "maturation pattern" (with HMB-45 and anti-tyrosinase) and a low proliferative index (with Ki-67). PMID- 11493378 TI - Bullous amyloidosis: the mechanism of blister formation revealed by electron microscopy. AB - BACKGROUND: Few electron microscopic studies of blister lesions in bullous amyloidosis have been reported, and the mechanism of blister formation remains to be elucidated. This study was designed to examine the nature of bullous amyloidosis ultrastructurally, and clarify the pathogenesis of blister formation. METHODS: We examined a 47-year-old woman with IgD-lambda type myeloma, suffering from bullous lesions on her hands and feet caused by trauma or rubbing. Light and electron microscopic studies were performed. RESULT: Ultrastructurally, amyloid deposits aggregated under the lamina densa. Keratinocyte protrusions penetrated the dermis through the gap in the lamina densa and enfolded amyloid deposits. Amyloid globules were found in enlarged intercellular spaces of keratinocytes. Desmosomes were sparsely distributed in some areas of the epidermis. CONCLUSION: These results indicate that keratinocytes enfold the amyloid globules and take them in the intercellular space of epidermis, and that the breakdown of the lamina densa and widening of the intercellular space between keratinocytes induce skin fragility. Trauma or rubbing of her hands and feet appears to act as the localized precipitating factor of blister formation in bullous amyloidosis. PMID- 11493379 TI - New insights into the applicability of T-cell receptor gamma gene rearrangement analysis in cutaneous T-cell lymphoma. AB - BACKGROUND: Detection of clonal T-cell receptor (TCR) gamma gene rearrangement by polymerase chain reaction (PCR) based method is a marker for cutaneous T-cell lymphoma (CTCL) although it can be seen in some benign dermatoses. To determine the accuracy of histologic criteria alone as well as the adjuvant diagnostic role of TCR gene rearrangement for the diagnosis of CTCL, we studied 100 patients with cutaneous T-cell infiltrates by both histology and TCR gene rearrangement. METHODS: The histologic features of the 100 patients were first reviewed by two independent dermatopathologists and their confidence in the diagnosis of CTCL was assigned one of four levels. Then the specimens were analyzed for TCR gene rearrangement either on paraffin-embedded or fresh-frozen tissue by PCR/denaturing gradient gel electrophoresis (DGGE). RESULTS: The clonality was detected in 100% (15/15) diagnostic of, 84.6% (11/13) consistent with, 57.6% (19/33) suggestive of CTCL. In 9 cases TCR gene rearrangement was compared between formalin-fixed and fresh specimens of the same individual, but with different degrees of histologic confidence (no lower than suggestive). In all cases fresh specimens were positive. In 5 of the cases (2-diagnostic, 2 consistent, 1-suggestive) formalin-fixed specimens were positive as well, and in 4 cases (1-consistent, 3-suggestive) formalin-fixed specimens were negative. When TCR gene rearrangement was studied in eight cases on sequential biopsies from the same patient, the clonality was detected in only one or two biopsies in four cases in which the histologic confidence was low (suggestive or nondiagnostic). The TCR gene rearrangement study showed identical banding patterns in lesions from different clinical stages in most patients. However, we observed that in one case, oligoclonal-banding pattern was seen in initial biopsy with histopathologic consistent with CTCL, while monoclonal banding pattern in more advanced lesion. CONCLUSIONS: Our data have demonstrated that TCR gene rearrangement studies by PCR/DGGE are consistently positive regardless of tissue fixation (formalin-fixed, paraffin-embedded vs. fresh-frozen tissue) and biopsy site when the histologic degree of confidence is very high (diagnostic). So, it may be of less importance as an adjuvant to histopathologic diagnosis for the cases with diagnostic CTCL histology. However, TCR gene rearrangement studies are particularly important in earlier cases with less conclusive histology, which provides strong confirmatory evidence of an evolving CTCL. In these cases, multiple biopsies may be required to establish the diagnosis and analysis of fresh tissue is suggested to increases the sensitivity. Moreover, our observation also suggested that some CTCL might not be monoclonal de novo, but oligoclonal instead. PMID- 11493380 TI - Granulomatous lymphangitis of the scrotum and penis. Report of a case and review of the literature of genital swelling with sarcoidal granulomatous inflammation. AB - BACKGROUND: Acquired lymphedema of the genitalia is a rare childhood presentation and is more common in elderly individuals secondary to pelvic/abdomenal malignancy or its therapy or worldwide due to filariasis. OBJECTIVE: Herein, we report a case of a healthy 11-year-old boy who presented with a 1-year history of chronic, asymptomatic scrotal and penile swelling. Biopsy revealed edema, lymphangiectases and peri- and intralymphatic sarcoidal type granulomas. This histologic pattern of granulomatous lymphangitis is most commonly associated with orofacial granulomatosis (granulomatous cheilitis and Melkersson-Rosenthal syndrome) and Crohn's disease. Treatment with topical steroids and physical support has resulted in marked improvement. No systemic disease (Crohn's disease) is evident 1 year later. Literature review revealed 44 cases of genital lymphedema with non-infectious granulomas. The majority of these young patients had Crohn's disease, frequently with anal involvement and a minority, both with and without Crohn's disease, had orofacial granulomatosis. CONCLUSIONS: Granulomatous lymphangitis should be considered in the differential diagnosis of chronic idiopathic swelling of the genitalia, particularly in younger individuals. Further clinical examination, additional laboratory studies and close follow-up for co-existing or subsequent development of Crohn's disease should be performed. The overlap between granulomatous lymphangitis of the genitalia, Crohn's disease and orofacial granulomatosis suggest that granulomatous lymphangitis of the genitalia may represent a forme fruste of Crohn's disease. PMID- 11493381 TI - Metastatic adenocarcinoma of the esophagus to the skin: new patterns of tumor recurrence and alternate treatments for palliation. AB - BACKGROUND: Esophageal cancer, particularly adenocarcinoma of the esophagus (ACE), has been steadily increasing in incidence in the United States. In the past, patients usually died rapidly with locoregional disease that leads to inanition and aspiration. However, today when patients with ACE are treated successfully with induction chemotherapy and radiation therapy, followed by surgical excision, ACE usually does not recur locally, but presents with metatastic disease. We present a 62-year-old white male with ACE, which was treated with induction chemotherapy and radiation therapy followed by surgical excision. After approximately 1 year with no evidence of locoregional recurrence, the patient presented with diffuse cutaneous metastatic disease. METHODS: In addition to routine staining immunohistochemical stains for CK(AE1/AE3), CK7, CK 20, EMA, alpha-smooth muscle (SM) actin, S-100 protein, CD34, P53, Bcl-2, c-erbB 2 were performed. RESULTS: The immunohistochemical profile was consistent with an esophageal origin showing positive staining with CK20 and CK7 as well as AE1/AE3 and EMA. In addition, there was marked nuclear expression of p53, as well as membrane expression of c-erb-B2; consistent with progression of the disease and poor response to further cytotoxic therapeutic regimes. CONCLUSIONS: With new therapeutic protocols, we can expect to see more metastatic disease with recurrences of ACE. The histopathologic features and the immunohistochemical profile of the recurrent tumors may be helpful in determining alternate forms of therapy that either alone or in combination could be useful in palliation and delaying progression. PMID- 11493382 TI - Cutaneous involvement by angioimmunoblastic T-cell lymphoma with remarkable heterogeneous Epstein-Barr virus expression. AB - INTRODUCTION: Initially described as an abnormal immune reaction, most cases of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD)-like T-cell infiltrates are now regarded as a peripheral T-cell lymphoma (AILD T-NHL). AILD T NHL is characterized clinically with constitutional symptoms, generalized lymphadenopathy, hepatosplenomegaly, skin rash, and polyclonal hypergammaglobulinemia. Epstein-Barr virus (EBV) is frequently detected in involved lymph nodes, but the presence of EBV in cutaneous infiltrates of AILD T NHL has rarely been examined. We present a patient with AILD T-NHL with cutaneous involvement that shows marked heterogeneity of EBV expression in the lymph node and skin biopsies, and review the histological findings of AILD T-NHL in the skin. METHODS: Two skin biopsies of a diffuse maculopapular rash and a lymph node were examined and immunophenotyped. In situ hybridization for detection of EBV in the lymph node and skin biopsies was utilized. In order to attempt to delineate which lymphocytes were EBV positive, skin biopsies were dual labeled with CD3, CD45RO, CD20 and EBV. The skin biopsies and lymph node were submitted for gene rearrangement studies by polymerase chain reaction (PCR). Capillary electrophoresis of fluorescently labeled PCR products was utilized for PCR product quantitation. RESULTS: The histological features of the lymph node were diagnostic of AILD T-NHL and a T-cell clone was identified by PCR. The skin biopsies showed an atypical superficial and deep perivascular polymorphous infiltrate consistent with cutaneous involvement by AILD T-NHL. Both skin biopsies showed the same clonal T-cell receptor gene rearrangement as the lymph node. In situ hybridization of the lymph node and one skin biopsy showed a few scattered EBV-positive lymphocytes (<1% of the infiltrate). A second skin biopsy revealed 40-50% of the lymphocytes as EBV positive. Dual staining for CD20 and EBV identified a minority of EBV-infected lymphocytes as B-cells, but most of the EBV-positive cells lacked staining for CD3 and CD45RO. CONCLUSIONS: In our patient, the same T-cell receptor gene rearrangement was found by PCR in all three biopsy sites. Most cases of AILD T-NHL contain only a few EBV-positive cells, but in our patient the extent of EBV expression ranged from <1% to 40-50% of the AILD T-NHL cutaneous infiltrate. To our knowledge, this case is the most extensive and heterogeneous expression of EBV in cutaneous AILD T-NHL to date. PMID- 11493383 TI - Ossified cylindroma in familial cylindromatosis (Brooke-Spiegler syndrome). PMID- 11493384 TI - Dietitians are constantly faced with new challenges and opportunities as new horizons emerge from our increased knowledge and understanding of dietetics. PMID- 11493385 TI - Validation of a nutrition screening tool: testing the reliability and validity. AB - BACKGROUND: The aim of this study was to validate a nutrition screening tool for use in South Manchester University Hospitals Trust. METHOD: A sample of 100 patients was selected from medical, surgical and elderly care wards. To test the reliability of the screening tool, nurses and dietitians completed the screening tool on the same patient. These results were compared for interobserver error to determine whether the screening tool was reproducible with different observers. To ascertain if the screening tool identified malnutrition at ward level, four markers commonly used to assess nutritional status were collected. These included body mass index (BMI), mid upper arm circumference MUAC, percentage weight loss, and energy intake calculated from the patient's first full day in hospital and expressed as a percentage of their estimated average requirements (EAR). RESULTS: There was a 95% level of agreement between nurses and dietitians within +/-3. The screening tool had a sensitivity level of 78% and a specificity of 52% when compared to all patients who had one or more markers indicating malnutrition. This association was found to be statistically significant (P < 0.005). CONCLUSION: The screening tool is reliable when completed by different observers and is valid for wide scale nutritional assessment. The screening tool identifies an acceptable number of patients who are malnourished but overestimates patients at moderate risk. PMID- 11493386 TI - Women's readiness to follow milk product consumption recommendations: design and evaluation of a 'stage of change' algorithm. AB - OBJECTIVE: To investigate readiness to follow milk product consumption recommendations in two random samples of New Zealand women, using an algorithm designed and evaluated to assess the stage of change construct of the Transtheoretical Model. PROTOCOL: Women were classified according to stage of readiness to perform two goal behaviours: consumption of two or four servings of milk products per day. Stage classification, as determined by the algorithm, was evaluated by estimating mean calcium intake in each stage using a validated food frequency questionnaire. This was undertaken in two independent samples of women randomly selected from the electoral rolls of two cities in New Zealand's South Island. RESULTS: Over 60% of women were classified as maintaining an intake of two servings of milk products per day. Of those women not meeting the goal of two servings per day, 73% had no intention of increasing their consumption. Over 80% of women were in the precontemplation stage for consuming four servings of milk products per day. Mean calcium intakes were significantly higher in women classified in action and maintenance stages than in preaction stages of change for both goal behaviours. CONCLUSION: Of those women not currently meeting the recommendation for two servings of milk products per day, most are in precontemplation, a stage characterized by resistance to change. Use of the staging algorithm developed in this study makes possible the classification of women according to their readiness to change, and thus the provision of appropriate stage-tailored advice. PMID- 11493387 TI - Women's confidence to consume adequate servings of milk products: validation of a summated rating scale. AB - OBJECTIVE: To design and validate a scale to assess women's confidence to consume adequate servings of milk products. DESIGN: Using the electoral rolls as a sampling frame, two large random population surveys of women aged 25-70 years were conducted. The structure of the self-efficacy scale was explored in the first survey of 500 women, and the component structure was refined and confirmed in an independent sample of 1200 women. RESULTS: The final single component self efficacy scale consisted of nine items to assess confidence to consume milk products in various situations. Confidence was higher in women who were already consuming adequate servings of milk products than in women who were not yet ready to consume adequate servings of milk products, providing construct validity for the scale. CONCLUSION: The data provide preliminary support for the validity of the self-efficacy scale. The potential usefulness of the scale in interventions designed to improve women's milk product consumption is discussed. PMID- 11493388 TI - Dietetic guidelines: diet in secondary prevention of cardiovascular disease. AB - AIM: To produce dietetic guidelines summarizing the systematic review evidence on dietary advice for secondary prevention of cardiovascular disease. METHODS: The Cochrane Library, MEDLINE and EMBASE were comprehensively searched for systematic reviews on aspects of diet and heart health. Reviews were included if they searched systematically for randomised controlled trials relating to diet and secondary prevention of cardiovascular disease. Two members of the UK Heart Health and Thoracic Dietitians Group critically appraised each review. The quality and results of each review were discussed and summarized in a whole group meeting. The guidelines were trialled in seven dietetic departments and modified where necessary. RESULTS: Providing evidence-based dietary information (including increasing omega-3 fat plus fruit and vegetable intake) to all people after myocardial infarction will save more lives than concentrating dietary advice on those in need of weight loss or lipid lowering. The practice of prioritizing dietetic time to those with raised lipids is out of date since the advent of statin therapy. CONCLUSION: There is good systematic review evidence that dietary advice to those with cardiovascular disease can reduce mortality and morbidity as well as modify risk factors. Dietary advice that does this most effectively should be prioritized. PMID- 11493389 TI - Survey of UK dietetic departments: diet in secondary prevention of myocardial infarction. AB - AIM: To collate information on current British dietetic practice in the area of diet in secondary prevention of myocardial infarction, and to compare this with best evidence. METHODS: A questionnaire was sent to all Chief Dietitians, to be completed by the whole department, asking about current dietary advice and practice, referrals and written dietary information for people following myocardial infarction. RESULTS: Fifty-seven per cent of questionnaires were returned. Dietary fat advice was prioritized by 84% of departments, fruit and vegetables by 49%, oily fish by 45% and fibre by 28%. Most dietitians (81%) felt that this advice would protect from further cardiovascular disease. Percentages of post-myocardial infarction patients given advice by a dietitian varied (median 60%), reasons for non-referral included patients having normal lipids or weight. More were given dietary advice by another health professional (median 90%); however, most dietitians were unsure about the quality of this advice. Diet sheets were felt to reflect advice priorities, but where sheets were sent they were much less likely than departments to prioritize oily fish or fruit and vegetable advice. CONCLUSION: Dietetic practice for people following myocardial infarction is out of line with current best evidence. Almost half of departments correctly prioritize oily fish advice, but often only see patients with raised lipids or weight. This is not the most effective strategy for preventing deaths. PMID- 11493390 TI - The involvement, understanding and attitudes of dietitians towards research and audit. AB - AIM: To investigate the involvement of dietitians in research and audit and to assess their understanding of and attitudes towards these activities. METHODS: A postal questionnaire was used to investigate the knowledge, attitude towards and involvement in research and audit of State Registered Dietitians practising in the National Health Service (NHS) in the UK. RESULTS: A response rate of 64% was achieved. Few respondents (15%) were currently involved in research, although more (65%) were currently involved in audit. Involvement in audit rose significantly with increasing grade and managerial responsibility. Dietitians qualifying more recently were more likely to believe that their college training prepared them to undertake research, but felt less confident about undertaking audit. Experience of audit made respondents more confident about auditing their work and improved their belief that they understood audit terminology. Higher grade dietitians were more likely to see audit as an important part of their role than were their more junior colleagues. Understanding of the terms 'research' and 'audit' was poor, although 48% of respondents identified the link between the two activities. Perceived constraints to involvement in research and audit were identified. CONCLUSION: The present level of involvement of dietitians in research and audit activities falls below that recommended in the BDA's (1997a) National Professional Standards for Dietitians Practising in Healthcare. This study identifies factors that influence dietitians' involvement in, understanding of and attitudes towards research and audit and recommends ways of increasing their involvement in these activities. PMID- 11493392 TI - Editorial. PMID- 11493395 TI - Infectious complications of lung transplantation. AB - Lung transplantation is associated with a high incidence of infection which directly impacts the morbidity and mortality associated with the procedure. In addition, these infections may also have immunologic consequences that play a role in the evolution of lung injury syndromes, resulting in earlier loss of graft than otherwise would be expected to occur. Although bacteria are responsible for the majority of infections following lung transplantation, fungal infections are associated with the highest mortality. This paper is an overview of the major infectious complications encountered in the lung transplant population. The epidemiology, prophylaxis, and treatment of infections following lung transplantation are critical areas for continued research. PMID- 11493396 TI - Community respiratory virus infections following lung transplantation. AB - Respiratory infections remain a significant cause of morbidity and mortality after lung transplantation. In addition to cytomegalovirus, the community respiratory viruses such as respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus, and adenovirus, are important causes of infection in transplant recipients, often involve the lower respiratory tract, and may be associated with significant morbidity and mortality. In this review, we summarize the current state of knowledge regarding the epidemiology, clinical manifestations, diagnosis, treatment and outcomes associated with RSV, PIV, influenza virus, and adenovirus infections in lung transplant recipients. PMID- 11493397 TI - Burkholderia cepacia complex: a contraindication to lung transplantation in cystic fibrosis? AB - Previous studies have indicated that pulmonary infection with Burkholderia cepacia is associated with poor clinical outcome after lung transplantation in cystic fibrosis (CF). Many treatment centers consider B. cepacia infection an absolute contraindication to lung transplantation. However, the B. cepacia complex actually consists of several closely related bacterial species. Although each of these has been isolated from CF sputum culture, certain species are much more frequently recovered than others, and it is not yet clear whether all species have the same potential for virulence in CF. Additional study is needed to better define the relative risks associated with each species of the B. cepacia complex. PMID- 11493398 TI - Aspergillosis in lung transplantation: incidence, risk factors, and prophylactic strategies. AB - Invasive aspergillosis remains a significant cause of morbidity and mortality in transplantation, especially lung and allogeneic bone marrow transplant recipients. The epidemiology, classic and newly recognized risk factors, and incidence of aspergillosis are reviewed. Risk factors include environmental exposures, airway colonization, profound immunosuppression, neutropenia, prior cytomegalovirus infection, and renal dysfunction. Clinical and radiographic presentations of invasive aspergillosis are discussed, including some unusual manifestations in lung transplant recipients. Early and accurate diagnosis of aspergillosis remains a challenge, and diagnostic strategies are reviewed, with an emphasis on the chest computerized tomography scan and on transbronchial or open lung biopsy. Recent advances include prophylactic and pre-emptive antifungal strategies, newer therapeutic agents, and improved risk stratification. PMID- 11493399 TI - The role of neutrophils in the pathogenesis of obliterative bronchiolitis after lung transplantation. AB - Obliterative bronchiolitis (OB) represents the most important long-term complication after lung transplantation. Elevated numbers of neutrophils within the airways are a hallmark of OB. It is unclear what causes the recruitment and activation of neutrophils in the airways of patients with OB: the process of chronic rejection itself or infection, which may (especially in latent virus infection) often be overlooked by the currently applied diagnostic procedures. It is well known that besides their physiologic functions in the clearance of invading micro-organisms, activated neutrophils have a remarkable potential to cause damage to lung tissue. This is attributable to their capability to generate reactive oxygen species and to release potentially toxic proteases. It has been shown that the increased numbers of neutrophils in bronchoalveolar lavage fluid of patients with bronchiolitis obliterans syndrome (BOS) after lung transplantation are associated with elevated levels of interleukin-8, the predominant neutrophil chemotactic factor in the lung. As evidence for the impact of neutrophils on the pathogenesis of BOS, there is significant oxidative stress within the airways of patients with BOS. In addition, the milieu within the airways is characterized by an imbalance between neutrophil elastase (NE) and molecules that inhibit NE as a result of an increased burden of NE released by neutrophils. A defective antiprotease shield due to the loss of secretory leukoprotease inhibitor could be demonstrated in BOS. These mechanisms may provide possible targets to develop new therapeutic strategies that either prevent neutrophil sequestration and activation, or inhibit neutrophil products in order to prevent or attenuate airway damage. PMID- 11493400 TI - Epstein--Barr virus post-transplant lymphoproliferative disease and virus specific therapy: pharmacological re-activation of viral target genes with arginine butyrate. AB - Lymphoproliferative disorders associated with the Epstein-Barr virus (EBV) include non-Hodgkin's lymphoma, Hodgkin's lymphoma, and "post-transplant lymphoproliferative disorders" (PTLD), which occur with immunosuppression after marrow and organ transplantation. PTLD is characterized by actively proliferating, latently infected EBV(+) B-lymphocytes, and often manifests a rapidly progressive fatal clinical course if the immunosuppression cannot be reversed. Lung transplant recipients are a subset of patients at special risk for developing PTLD. The incidence of PTLD development in these patients has been estimated at 5--10%. Whereas immunologic and antiviral therapy have been moderately effective for treating EBV-associated infections in the lytic phase, they have been less useful in the more common latent phase of the disease. One common treatment for herpesvirus infections has targeted the virus-specific enzyme thymidine kinase (TK). The lack of viral TK expression in EBV(+) tumor cells, due to viral latency, makes anti-viral therapy alone ineffective as an anti-neoplastic therapy, however. We have developed a strategy for the treatment of EBV-associated lymphomas/PTLD using pharmacologic induction of the latent viral TK gene and enzyme in the tumor cells, followed by treatment with ganciclovir. Arginine butyrate selectively activates the EBV TK gene in latently EBV-infected human lymphoid cells and tumor cells. A Phase I/II trial has been initiated, employing an intra-patient dose escalation of arginine butyrate combined with ganciclovir. In six patients with EBV-associated lymphomas or PTLD, all of which were resistant to conventional radiation and/or chemotherapy, this combination produced complete clinical responses in four of six patients, with a partial response occurring in a fifth patient. Pathologic examination in two of three patients demonstrated complete necrosis of the EBV lymphoma, with no residual disease, following a single three-week course of the combination therapy. Possible side-effects of the therapy included nausea and reversible lethargy at the highest doses. One patient suffered acute liver failure, thought to be secondary to release of FasL from the necrotic tumor. Analysis of patient derived tumor cells in culture demonstrated that arginine butyrate produced selective induction of the EBV TK gene, which then conferred sensitivity to ganciclovir, resulting in tumor apoptosis. Additional patient accrual is sought for further evaluation of this therapy. PMID- 11493401 TI - Commercial dry dog food in the north central United States is not contaminated by Dermatophagoides house dust mites. AB - Contamination of home-stored cereal grain food products with Dermatophagoides spp. house dust mites (HDM) was reported recently, along with anaphylaxis after consumption of these foods by dust mite-allergic people. We hypothesized that commercial dry dog food could become similarly contaminated, particularly if stored improperly, and could eventually contribute to allergic signs in dogs. Newly purchased bags of dry dog food (n = 30), from a variety of sources and manufacturers, and client samples of dry dog food (n = 50), stored under a variety of conditions, were obtained. Food samples were extracted in aqueous buffer, and extracts were assayed using ELISA for Dermatophagoides group II (Der II) allergen, as a marker for the presence of HDM. Der II allergen was not detected in any of the 30 newly purchased or 50 stored samples tested. Positive control samples consisting of house dust or dog food mixed with house dust, similarly extracted, and Dermatophagoides commercial allergen extract were positive for Der II in the same assay. We could find no evidence of HDM contamination in newly purchased or stored commercial dry dog food in the north central United States. PMID- 11493402 TI - Double-blinded, placebo-controlled, cross-over pilot study on the efficacy of zileuton for canine atopic dermatitis. AB - Nine dogs meeting the diagnostic criteria for canine atopic dermatitis were enrolled in a double-blind, placebo-controlled, cross-over clinical trial. In this pilot study, zileuton (a 5-lipoxygenase inhibitor) given orally at 2 mg kg( 1) three times daily for 4 weeks significantly decreased erythema in dogs with atopic dermatitis but had no effect on pruritus. Zileuton was well tolerated and no adverse clinical signs were noted. However, one dog developed mild alanine aminotransaminase elevation, which resolved within 1 week of discontinuation of therapy. Monitoring of alanine aminotransaminase may be necessary in dogs receiving zileuton. Further studies with larger number of dogs are needed to evaluate the efficacy of zileuton as treatment for canine atopic dermatitis. PMID- 11493403 TI - Adverse reactions to FMD vaccine. AB - A case of adverse post-vaccination allergic reactions, which occurred in a dairy cattle herd 8 days after the annual foot-and-mouth disease (FMD) vaccination, is described. The dermatologic lesions observed in these cattle included: urticaria, exudative and necrotic dermatitis, along with oedema and vesicles on the teats. These reactions occurred in 11.3% of the heifers, in 10% of the first-lactating cows and in 14.6% of the adult cows. The average loss of milk production for an affected cow on this farm was 21.5% per day, for seven consecutive days. The extent of the lesions was apparently related to concurrent diseases such as bovine virus diarrhea - mucosal disease complex (BVD-MD) and Johne's disease and, to a lesser degree, correlated with the age or breed. PMID- 11493404 TI - Two cases of dermatophytic pseudomycetoma in the dog: an immunohistochemical study. AB - Two cases of canine dermatophytic pseudomycetoma resulting in subcutaneous nodules resembling those previously reported in Persian cats are described. Culture performed from one nodule yielded dark yellow colonies consistent with Microsporum canis. Immunohistochemistry, using rabbit anti-M. canis, demonstrated specific binding to fungal elements in paraffin sections. The specificity of the antiserum was further tested by an agar gel immunodiffusion assay using a soluble extract from a feline isolate of M. canis as antigen. The antiserum did not cross react with an Aspergillus fumigatus antigen. These are the first two reported cases of canine dermatophytic pseudomycetoma and immunohistochemical staining supported the diagnosis. PMID- 11493406 TI - Congenital hypotrichosis in a Percheron draught horse. AB - A blue roan Percheron foal was born with poorly circumscribed patchy alopecia of the trunk and legs. Teeth and hoof development were normal. Alopecia was progressive, becoming almost complete by 1 year of age. Histopathological findings in a skin biopsy obtained at 7 months of age were consistent with severe follicular hypoplasia. Sebaceous glands, epitrichial sweat glands and arrector pilae muscles were normal. The horse is alive and otherwise well at 6 years of age, although adult stature is considered small for this breed. The clinical history and histopathological findings are most consistent with a form of congenital hypotrichosis. PMID- 11493405 TI - Idiopathic linear pustular acantholytic dermatosis in a young Brittany Spaniel dog. AB - A unilateral, linear, papular-pustular dermatosis is described in a young Brittany Spaniel dog. The dermatosis appeared to follow Blaschko's lines and extended from the left inguinal region to the medial aspect of the left metatarsal area. The predominant histological finding was an eosinophilic and neutrophilic pustular mural folliculitis with prominent acantholysis of infundibular epithelium. There was a rapid and long-lasting (> 15 months) resolution after oral administration of methylprednisolone (1.6 mg kg(-1)). PMID- 11493407 TI - Zonal dermal separation: a distinctive histopathological lesion associated with hyperelastosis cutis in a Quarter Horse. AB - This case report describes a distinctive deep cutaneous lesion in a 1-year-old Quarter Horse filly with hyperelastosis cutis. The horse had a typical clinical presentation of hyperelastic skin associated with a 6-month history of cutaneous wounds that developed following minor cutaneous trauma. Punch biopsies of skin from the affected horse were thinner than similar biopsies from an age- and breed matched control. Significant microscopic lesions were not seen in cutaneous punch biopsies stained with haematoxylin and eosin and Masson's trichrome stains, but the ultrastructure of the dermis from the affected horse was characterized by variation in collagen fibre diameter and loose packing of collagen fibres within bundles. The horse was euthanized and necropsied, and full-thickness sections of skin were collected and examined microscopically. Affected skin was of normal thickness; however, the deep dermis contained a distinctive horizontal linear zone in which separation of collagen bundles resulted in the formation of large empty cleft-like spaces between the upper and lower regions of the deep dermis. We suggest the term 'zonal dermal separation' for this microscopic lesion. Incisional full-thickness skin biopsies should be taken in suspected cases of equine hyperelastosis cutis because punch biopsies may not obtain enough deep dermis to adequately represent pathological change in the skin of horses with this disorder. PMID- 11493408 TI - In vitro activity of fluoroquinolones (gatifloxacin, levofloxacin and trovafloxacin) and seven other antibiotics against Streptococcus pneumoniae. AB - In recent years, the level of resistance of S. pneumoniae to beta-lactam and/or macrolides has increased around the world including some countries in South America. Because of this resistance, it is necessary to test the therapeutic alternatives for treating this pathogen, including the newer quinolones. This study was carried out in order to compare the in vitro activity of fluoroquinolones gatifloxacin, levofloxacin and trovafloxacin, to penicillin G, amoxicillin, amoxicillin-clavulanate, cufuroxime sodium, ceftriaxone, azithromycin and clarithromycin, against 300 strains of S. pneumoniae. Of the 300 samples tested, 18.6% were not susceptible to penicillin (56 strains) and 7% (21 strains) were resistant to the second generation cephalosporin. Among the macrolides, resistance ranged from 6.7% for clarithromycin to 29.6% for azithromycin. Susceptibility to the newer quinolones was 100% including the 56 strains not susceptible to penicillin. Among the 10 antibiotics evaluated, the fluoroquinolones gatifloxacin, levofloxacin, and trovafloxacin displayed high levels of in vitro activity against S. pneumoniae. PMID- 11493409 TI - Low-dose adrenocorticotropin test in patients with the acquired immunodeficiency syndrome. AB - Adrenocortical insufficiency is a serious complication of AIDS. Usually, integrity of the hypothalamo-pituitary-adrenal (HPA) axis in AIDS patients is assessed by measuring basal cortisol levels and cortisol response to 250 microg of ACTH. Recent studies suggest that a lower ACTH dose increases the sensitivity of the procedure. In the present study, we investigated the prevalence of adrenal hypofunction in AIDS patients using a low-dose ACTH test (1 microg), evaluated the clinical characteristics that might suggest this diagnosis, and the diseases and/or drugs that could be associated with it. We prospectively evaluated 63 very ill AIDS patients and 16 normal controls. A standard examination assessed the presence of signs and symptoms of adrenal insufficiency. Blood samples were collected before and 30 and 40 minutes after an injection of 1 microg 1-24 ACTH. No opportunistic disease, signs, symptoms or drugs were associated with an abnormal cortisol response to ACTH. The lowest stimulated cortisol level in the control group was 18.5 microg/dL; cortisol levels > or = 18 microg/dL were taken to indicate a normal HPA axis. Test results revealed that 12/63 AIDS patients (19%) had an abnormal HPA axis. With these data in mind, we suggest a prospective adrenal function evaluation of all severely ill AIDS patients. PMID- 11493410 TI - Evaluation of efficacy and safety of itraconazole oral solution for the treatment of oropharyngeal candidiasis in aids patients. AB - This study was a non-comparative multicenter clinical trial to evaluate the efficacy and tolerability of itraconazole oral solution 200 mg/day (100 mg twice a day in the fasting state) for the treatment of oropharyngeal candidiasis in AIDS patients. We included 50 patients who were treated and followed for up to 3 weeks after ending therapy in the analysis. Mycological cures at the end of therapy occurred in 20/50 patients (40%), but colonization by Candida sp. was recorded in 42/50 (84%) by the end of follow-up. A high rate of clinical response was observed in 46/50 (92%), and the response was sustained for up to 21 days after stopping therapy in 24/46 patients (52%). Clinical relapses were documented among 22 patients, but all causative fungal organisms associated with a relapse were susceptible to itraconazole. There were many patients with persistence or recurrence of Candida, but without mucositis. Relapse of Candida mucositis was significantly related to low levels of CD(4) lymphocytes exhibited by symptomatic patients. The drug was well tolerated by all but 1 patient. We conclude that itraconazole oral solution (100 mg bid for 7-14 days) is a well tolerated and effective treatment for suppressing the symptoms of oropharyngeal candidiasis in AIDS patients. Patients with severe immunosuppression may relapse and require frequent cycles of treatment or longterm suppressive therapy. PMID- 11493411 TI - Atovaquone and proguani hydrochloride compared with chloroquine or pyrimethamine/sulfodaxine for treatment of acute Plasmodium falciparum malaria in Peru. AB - The efficacy and safety of a fixed-dose combination of atovaquone and proguanil hydrochloride (Malarone) were compared with chloroquine or pyrimethamine/sulfadoxine in patients with acute falciparum malaria in northern Peru. Patients were initially randomized to receive 1,000 mg atovaquone and 400 mg proguanil hydrochloride daily for 3 days (n=15) or 1,500 mg chloroquine (base) over a 3 day period (n=14) (phase 1). The cure rate with chloroquine was lower than expected and patients were subsequently randomized to receive a single dose of 75 mg pyrimethamine and 1,500 mg sulfadoxine (n=9) or atovaquone/proguanil as before (n=5) (phase 2). In phase 1, atovaquone/proguanil was significantly more effective than chloroquine (cure rate 100% [14/14] vs. 8% [1/13], P<0.0001). In phase 2, atovaquone/proguanil and pyrimethamine/sulfadoxine were both highly effective (cure rates 100% [5/5] and 100% [7/7]). There were no significant differences between treatment groups in parasite or fever clearance times. Adverse events were typical of malarial symptoms and did not differ significantly between groups. Overall efficacy of atovaquone/proguanil was 100% for treatment of acute falciparum malaria in a region with a high prevalence of chloroquine resistance. PMID- 11493412 TI - Chest radiograph abnormalities in patients hospitalized with leptospirosis in the city of Salvador, Bahia, Brazil. AB - This study was designed to estimate the prevalence of pulmonary radiograph abnormalities and describe the distribution of the patterns of radiographic alterations among patients hospitalized with leptospirosis. Chest radiographs of 139 patients hospitalized with leptospirosis in Couto Maia Hospital, in Salvador, Bahia, Brazil, between July, 1997, and July, 1999, were analyzed. The radiographs were requested soon after hospital admission, independent of the clinical manifestations of the patients. Only the first radiograph was considered. Pulmonary radiograph alterations were recorded in 35/139 patients (25.2%); 95% mid-point confidence interval = 18.5% to 32.9%. Among the patients with radiograph alterations, alveolar infiltrate was seen in 26/35 (74.3%). The lesions were bilateral in 54.3% and located in the inferior lobes in 45.5%. Pleural effusion, represented by blunting of the costo-phrenic angle, was detected in 8.6% of the patients. The pattern of the pulmonary alterations, predominantly bilateral alveolar infiltrates, is consistent with the evidence that the basic pulmonary alteration in leptospirosis is a generalized capillaritis. PMID- 11493413 TI - Successful prevention of hiv transmission from mother to infant in Brazil using a multidisciplinary team approach. AB - OBJECTIVES: To determine the HIV vertical transmission rate (VTR) and associated risk factors by use of zidovudine and infant care education in Brazil. METHODS: Since 1995, a prospective cohort of HIV infected pregnant women has been followed at the Federal University of Rio de Janeiro. A multidisciplinary team was established to implement the best available strategy to prevent maternal-infant HIV transmission. Patients with AIDS or low CD(4) and high viral load received anti-retroviral drugs in addition to zidovudine. Children were considered infected if they had 2 positive PCR-RNA tests between 1 and 4 months of age, or were HIV antibody positive after 18 months. Education regarding infant treatment and use of formula instead of breast feeding was provided. RESULTS: Between 1995 and August, 2000, HIV status was determined for 145 infants. Compliance with intra-partum treatment, infant treatment and use of formula was 88.2%. Intra partum zidovudine treatment was completed in 134/145 (92.6%) of patients; 88.1% had rupture of membranes < 4 hours; 85.4% of mothers were asymptomatic. The mean CD(4) count was 428.4 cells and mean viral load 39,050 copies. HIV vertical transmission rate was 4/145 (2.75%; CI: 0.1%-5.4%). The only risk factor significantly associated with transmission was a failure to use zidovudine intra partum in 2 of the 4 mothers (50% versus 6.4% in non-transmitting mothers). A trend toward low CD(4) and high viral load at entry, and rupture of membranes > 4 hours were associated with increased HIV transmission. CONCLUSION: HIV vertical transmission in Brazil was reduced to a level similar to other countries with the most effective prevention programs using a multidisciplinary team approach. A high level of compliance for use of anti-retroviral drugs, the provision of health education to mothers, and use of formula for all exposed infants. PMID- 11493414 TI - Etiology of childhood community acquired pneumonia and its implications for vaccination. AB - Pneumonia is an important cause of morbidity and mortality among children throughout the world. Vaccines are available for some organisms, but they are underutilized and/or still in development. To evaluate the potential impact of vaccines, we review studies in which the etiology of childhood community-acquired pneumonia was recorded. In North America and Europe (9 studies), the etiology of pneumonia was established in 62% of studied children (range 43%-88%) by use of noninvasive specific methods for microbiologic diagnosis. The most often identified agents were S. pneumoniae (22%), respiratory syncytial virus (RSV) (20%), Haemophilus influenzae (7%), and Mycoplasma pneumoniae (15%). In Africa and South America (8 studies), bacteria were recovered from 56% (range 32%-68%) of severely ill children studied by lung aspirate. The most often isolated bacteria were Streptococcus pneumoniae (33%) and Haemophilus influenzae (21%). A high percentage of H. influenzae strains were not serotype b. Throughout the world, children requiring hospitalization were most likely to have infection caused by pneumococcus, H. influenzae or RSV. Out patients also had Mycoplasma pneumoniae. Countries in Africa and Asia recorded 2 to 10 times more children with pneumonia (7 to 40/100 annually) than in the USA. Widespread use of pneumococcal and H. influenzae type b conjugate vaccines could reduce the frequency of childhood pneumonia by one-third. Further reduction will require development of non-type b H. influenzae, RSV and M. pneumoniae vaccines. This could result in a > 50% reduction of pneumonia in children. This goal should be sought and achieved as soon as possible. PMID- 11493415 TI - Mycobacterial spindle cell pseudotumor of the appendix vermiformis in a patient with aids. AB - Mycobacterial pseudotumor (MP) is a rare pathologic presentation of both Mycobacterium tuberculosis and non-tuberculous mycobacterial disease, hitherto reported to occur only in immunosuppressed patients with or without human immunodeficiency virus infection. This lesion shares close pathologic resemblance to certain mesenchymal neoplasms, particularly Kaposi's sarcoma (KS), from which it must be properly differentiated due to distinct prognosis and therapy. We report a case of MP obliterating the lumen of the appendix vermiformis in a 34 year-old patient who died of complications of AIDS at our hospital in Rio de Janeiro. A total of 24 cases of MP (including our patient) have been described in the literature. MP has been found especially in lymph nodes, but extranodal lesions have been described in the skin, spleen, lung, bone marrow, brain and, in our patient, the appendix vermiformis. We offer a review of the other 23 published case reports of MP in both HIV-infected and uninfected patients and discuss the pathologic features that differentiate MP from KS. PMID- 11493416 TI - Preventive health care for an endangered species--the physician scientist. PMID- 11493417 TI - Breast imaging: mammography and ultrasonography. AB - Mammography is the primary imaging for breast cancer screening and diagnosis. Recent improvements in mammography are related to programs established by professional societies and government agencies to bring standardization and quality assurance in the technical improvements that have extended its contributions to breast imaging. This article addresses the current roles and limitations of mammography and ultrasonography in the detection and diagnosis of breast cancer. PMID- 11493418 TI - MR imaging of the breast. AB - The results of clinical investigation suggest that MR imaging can provide clinically important information that cannot be obtained with conventional imaging methods, and that this modality will, in the future, be an invaluable adjunctive breast imaging tool, just as breast imaging is today. MR imaging as a method to detect, diagnose, and stage breast cancer remains in the investigational stage, but is emerging as perhaps the most promising imaging modality for breast cancer detection. PMID- 11493419 TI - An overview of interpretation strategies for breast MR imaging. AB - This article provides an overview of the different approaches to the interpretation of breast MR imaging. It provides an overview of the use of architectural features for breast MR imaging interpretation. It also includes a discussion of the various types of contrast kinetic data that are used for breast MR imaging interpretation. Approaches to combine architectural and kinetic features are also discussed. This article serves as an introduction to the other articles in this issue that discuss specific interpretation and strategies. PMID- 11493420 TI - Progress report from the American College of Radiology Breast MR Imaging Lexicon Committee. AB - Given the wide variety of terms for abnormal findings on contrast-enhanced breast MR imaging studies, the United States Public Health Services' Office on Women's Health, the Susan G. Komen Foundation for Breast Cancer Research, and the American College of Radiology, funded a committee (or "working group") of international experts to develop a standard language for breast MR imaging reporting. During the last three years, the group members have developed a preliminary Breast MR Imaging Lexicon to describe abnormal contrast-enhancing findings in the breast on MR imaging studies, with a special focus on lesion morphology on high-resolution scans and kinetic curve data descriptions. The initial version of the Breast MR Imaging Lexicon has undergone preliminary testing and is undergoing further development and refinement based on test results. Future work on the new Breast MR Imaging Lexicon includes continued lexicon development, accumulation of a breast MR imaging case set for lexicon testing, and planning for subsequent education for general radiologists on the new lexicon. PMID- 11493421 TI - Architectural-based interpretations of breast MR imaging. AB - Architectural features extracted from high spatial resolution MR images of the breast can help distinguish malignant from benign breast abnormalities and can help predict specific histopathologic diagnoses. Various breast pathologies consistently display certain MR architectural features. Individual architectural features have been incorporated into interpretation models that can improve the diagnostic performance of MR imaging over the use of individual features alone. PMID- 11493422 TI - Vascularity assessment of breast lesions with gadolinium-enhanced MR imaging. AB - Gadolinium-enhanced MR imaging of the breast can be used to characterize both tumor morphology and vascularity. An empirical three-point method using high resolution three-dimensional MR imaging that combines high spatial resolution with an estimate of tumor pharmacokinetics is described. This technique appears to improve diagnostic specificity of breast MR imaging and may provide a noninvasive method of tumor characterization of prognostic value. PMID- 11493423 TI - Cancer staging with breast MR imaging. AB - Breast MR imaging is a useful tool for staging breast cancer patients, and staging is more accurate with MR imaging than with conventional imaging techniques. MR imaging is the preferred imaging test for the accurate staging of breast cancer before surgery and for assessment of patients with positive axillary adenopathy and negative mammogram and physical examination. There are many important questions regarding the role of MR imaging in breast cancer staging that must be addressed by future research and involvement of MR imaging of the breast in clinical trials. PMID- 11493424 TI - Potential role of magnetic resonance imaging and other modalities in ductal carcinoma in situ detection. AB - Ductal Carcinoma In Situ (DCIS) is the earliest form of ductal cancer, with a high rate of care if treated early. This article outlines the use of breast imaging in DCIS diagnosis, including mammography, MR imaging, and nuclear medicine studies. While MR imaging and nuclear medicine show great promise in DCIS diagnosis, mammography remains the mainstay of DCIS detection by the presence of microcalcifications in early tumors on the mammogram. PMID- 11493425 TI - MR imaging in screening women at increased risk for breast cancer. AB - Hereditary breast cancer accounts for 5% to 10% of the total breast cancer burden. Screening of this group of women has been done by palpation and conventional mammography until recently, but because of the age group, mammography has a limited value. MR mammography has been demonstrated to be a reliable imaging modality in this group of patients. PMID- 11493426 TI - MR imaging-guided interventions in the breast. AB - MR imaging of the breast has high sensitivity for the detection of invasive breast cancer. However, not all enhancing lesions are malignant. A needle localization or biopsy system is necessary to differentiate false positive benign enhancing lesions from the true carcinomas. In this article, the techniques, pitfalls, and potential clinical indications for MR imaging-guided needle localization and percutaneous biopsy are discussed. PMID- 11493427 TI - MR-guided minimally invasive procedures. AB - This article outlines the integration of breast MR imaging minimally invasive therapy for breast tumors. Technical obstacles discussed include accurate determination of margins, DCIS, and localization methods. Treatment methods such as cryotherapy, interstitial hyperthermia, and focused ultrasound are discussed. Other subjects include the amount of minimally-invasive therapy performed to date and the ethical dilemma of clinical trials. PMID- 11493428 TI - Imaging of the patient with silicone gel breast implants. AB - There are two reasons for radiologic evaluation of the augmented breast. Because women with implants are at the same risk for breast cancer as other women, imaging is performed to screen for cancer or to work up clinical abnormalities. Additionally, imaging allows assessment of implant integrity. The various methods for imaging implants and breast tissue in the augmented patient are discussed. Imaging findings suggestive of silicone gel implant rupture are presented. PMID- 11493431 TI - Granulocyte colony-stimulating factor regulates myeloid differentiation through CCAAT/enhancer-binding protein epsilon. AB - Granulocyte colony-stimulating factor (G-CSF) is a major cytokine that regulates proliferation and differentiation of myeloid cells, although the underlying mechanisms by which G-CSF controls myeloid differentiation are largely unknown. Differentiation of hematopoietic cells is regulated by lineage-specific transcription factors, and gene-targeting studies previously revealed the critical roles of CCAAT/enhancer-binding protein (C/EBP) alpha and C/EBP epsilon, respectively, in the early and mid-late stages of granulocyte differentiation. The expression of C/EBP epsilon in 32Dcl3 cells and FDCP1 cells expressing mutant G-CSF receptors was examined and it was found that G-CSF up-regulates C/EBP epsilon. The signal for this expression required the region containing the first tyrosine residue of G-CSF receptor. Dominant-negative signal transducers and activators of transcription 3 blocked G-CSF--induced granulocytic differentiation in 32D cells but did not block induction of C/EBP epsilon, indicating that these proteins work in different pathways. It was also found that overexpression of C/EBP epsilon greatly facilitated granulocytic differentiation by G-CSF and, surprisingly, that expression of C/EBP epsilon alone was sufficient to make cells differentiate into morphologically and functionally mature granulocytes. Overexpression of c-myc inhibits differentiation of hematopoietic cells, but the molecular mechanisms of this inhibition are not fully understood. In 32Dcl3 cells overexpressing c-myc that do not differentiate by means of G-CSF, induction of C/EBP epsilon is completely abrogated. Ectopic expression of C/EBP epsilon in these cells induced features of differentiation, including changes in nuclear morphologic characteristics and the appearance of granules. These data show that C/EBP epsilon constitutes a rate-limiting step in G-CSF-regulated granulocyte differentiation and that c-myc antagonizes G-CSF-induced myeloid differentiation, at least partly by suppressing induction of C/EBP epsilon. (Blood. 2001;98:897 905) PMID- 11493432 TI - Depletion of circulating natural type 1 interferon-producing cells in HIV infected AIDS patients. AB - Natural interferon-alpha producing cells (IPCs) are a newly characterized blood cell type, which is the major source of type I interferons in antiviral innate immune responses. The relationship between the number of circulating IPCs, HIV disease progression, and the occurrence of HIV-related complications was investigated. The study of 25 healthy donors and 54 HIV-infected subjects demonstrated a direct correlation between blood IPC number, interferon-alpha production, and clinical state of HIV-infected subjects. Asymptomatic long-term survivors had increased IPC number and function relative to uninfected controls and infected individuals with progressive disease. IPC numbers were markedly reduced in AIDS patients developing opportunistic infections and cancer. A negative correlation was found between the IPC number in the blood and the HIV viral load, suggesting that IPCs are important in controlling HIV replication. This study provides the first evidence that IPCs are being affected during the course of HIV infection and suggests that these cells can play a vital role in the protection against opportunistic pathogens and cancer. (Blood. 2001;98:906 912) PMID- 11493433 TI - Signaling pathways activated by daunorubicin. AB - The anthracycline daunorubicin is widely used in the treatment of acute nonlymphocytic leukemia. The drug has, of course, been the object of intense basic research, as well as preclinical and clinical study. As reviewed in this article, evidence stemming from this research clearly demonstrates that cell response to daunorubicin is highly regulated by multiple signaling events, including a sphingomyelinase-initiated sphingomyelin-ceramide pathway, mitogen activated kinase and stress-activated protein/c-Jun N-terminal kinase activation, transcription factors such as nuclear factor kappa B, as well as the Fas/Fas ligand system. These pathways are themselves influenced by a number of lipid products (diacylglycerol, sphingosine-1 phosphate, and glucosyl ceramide), reactive oxygen species, oncogenes (such as the tumor suppressor gene p53), protein kinases (protein kinase C and phosphoinositide-3 kinase), and external stimuli (hematopoietic growth factors and the extracellular matrix). In light of the complexity and diversity of these observations, a comprehensive review has been attempted toward the understanding of their individual implication (and regulation) in daunorubicin-induced signaling. (Blood. 2001;98:913-924) PMID- 11493434 TI - Blocking of c-FLIP(L)--independent cycloheximide-induced apoptosis or Fas mediated apoptosis by the CC chemokine receptor 9/TECK interaction. AB - Chemokines play a pivotal role in regulating leukocyte migration as well as other biological functions. CC chemokine receptor 9 (CCR9) is a specific receptor for thymus-expressed CC chemokine (TECK). It is shown here that engagement of CCR9 with TECK leads to phosphorylation of Akt (protein kinase B), mitogen-activated protein kinases (MAPKs), glycogen synthase kinase--3 beta (GSK-3 beta), and a forkhead transcription factor, FKHR, in a human T-cell line, MOLT4, that naturally expresses CCR9. By means of chemical inhibitors, it is shown that phosphoinositide-3 kinase (PI-3 kinase), but not MAPK, is required for CCR9 mediated chemotaxis. Akt, GSK-3 beta, FKHR, and MAPK have been previously implicated in cell survival signals in response to an array of death stimuli. When MOLT4 cells, which expressed Fas as well as CXCR4, were stimulated with cycloheximide (CHX), an agonistic anti-Fas antibody, or a combination of these, the cells rapidly underwent apoptosis. However, costimulation of MOLT4 cells with TECK or stromal derived factor--1 significantly blocked CHX-mediated apoptosis, whereas stimulation only with TECK partially blocked Fas-mediated apoptosis. Concomitant with this blocking, cleavage of poly (adenosine 5'-diphosphate- ribose) polymerase and activation of caspase 3 were significantly attenuated, but the expression level of FLICE inhibitory protein c-FLIP(L), which had been shown to be regulated by CHX, was unchanged. This demonstrates that activation of CCR9 leads to phosphorylation of GSK-3 beta and FKHR and provides a cell survival signal to the receptor expressing cells against CHX. It also suggests the existence of a novel pathway leading to CHX-induced apoptosis independently of c FLIP(L). (Blood. 2001;98:925-933) PMID- 11493435 TI - T-cell--depleted allogeneic bone marrow transplantation followed by donor lymphocyte infusion in patients with multiple myeloma: induction of graft-versus myeloma effect. AB - Previous trials of allogeneic bone marrow transplantation (BMT) in patients with multiple myeloma (MM) have demonstrated high response rates but also high transplantation-related mortality (TRM) and high relapse rates. Exploitation of this strategy remains of interest because donor lymphocyte infusions (DLIs) can induce a potent graft-versus-myeloma (GVM) effect. CD6 T-cell--depleted allogeneic BMT was combined with prophylactic CD4(+) DLI administered 6 to 9 months after BMT in an effort to reduce TRM and to induce a GVM response after BMT. Twenty-four patients with matched sibling donors and chemotherapy-sensitive disease underwent BMT. CD6 T-cell depletion of donor bone marrow was the sole method of graft-versus-host disease (GVHD) prophylaxis. GVHD after BMT was minimal, 1 (4%) grade III and 4 (17%) grade II GVHD. Fourteen patients received DLI, 3 in complete response and 11 with persistent disease after BMT. Significant GVM responses were noted after DLI in 10 patients with persistent disease, resulting in 6 complete responses and 4 partial responses. After DLI, 50% of patients developed acute (> or = II) or extensive chronic GVHD. Two-year estimated overall survival and current progression-free survival (PFS) for all 24 patients is 55% and 42%, respectively. The 14 patients receiving DLI had an improved 2-year current PFS (65%) when compared with a historical cohort of MM patients who underwent CD6-depleted BMT survived 6 months with no GVHD and did not receive DLI (41%) (P =.13). Although this study suggests that prophylactic DLI induces significant GVM responses after allogeneic BMT, only 58% of patients were able to receive DLI despite T-cell--depleted BMT. Therefore, less toxic transplantation strategies are needed to allow a higher proportion of patients to receive DLI and the benefit from the GVM effect after transplantation. (Blood. 2001;98:934-939) PMID- 11493436 TI - Lack of correlation between numbers of circulating t(14;18)-positive cells and response to first-line treatment in follicular lymphoma. AB - In follicular lymphoma, the t(14;18) status of the peripheral blood and bone marrow analyzed by polymerase chain reaction (PCR) is assumed to correlate with disease activity in patients with relapsed disease. The clinical significance of quantitating circulating lymphoma cells by real-time PCR is reported in patients on first-line treatment. Thirty-four consecutive patients with previously untreated follicular lymphoma and detectable t(14;18)-positive cells in pretreatment peripheral blood samples were monitored. All patients were treated with standard chemotherapy in combination with interferon alfa-2b. Before and after induction therapy, blood samples were taken for quantitative analysis of t(14;18). At presentation, a median of 262 t(14;18)-positive cells per 75,000 normal cells was found (range, 1-75 000). Patients with lower numbers of circulating tumor cells more frequently had bulky disease (P =.02). Seventy-nine percent of the patients responded clinically to treatment. In 22 of 28 patients, including 4 patients in whom treatment had failed clinically, the number of circulating t(14;18)-positive cells decreased to undetectable or low levels after therapy. In the remaining responding patients, circulating tumor cells persisted after therapy. These quantitative data on circulating t(14;18)-positive cells call into question the usefulness of molecular monitoring of the blood in a group of patients with follicular lymphoma uniformly treated with a noncurative first line regimen. T(14;18)-positive cells decreased in peripheral blood after treatment, irrespective of the clinical response. Therefore, the significance of so-called molecular remission should be reconsidered in follicular lymphoma. (Blood. 2001;98:940-944) PMID- 11493437 TI - Expression of a single gene, BCL-6, strongly predicts survival in patients with diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphoma (DLBCL) is characterized by a marked degree of morphologic and clinical heterogeneity. Establishment of parameters that can predict outcome could help to identify patients who may benefit from risk adjusted therapies. BCL-6 is a proto-oncogene commonly implicated in DLBCL pathogenesis. A real-time reverse transcription-polymerase chain reaction assay was established for accurate and reproducible determination of BCL-6 mRNA expression. The method was applied to evaluate the prognostic significance of BCL 6 expression in DLBCL. BCL-6 mRNA expression was assessed in tumor specimens obtained at the time of diagnosis from 22 patients with primary DLBCL. All patients were subsequently treated with anthracycline-based chemotherapy regimens. These patients could be divided into 2 DLBCL subgroups, one with high BCL-6 gene expression whose median overall survival (OS) time was 171 months and the other with low BCL-6 gene expression whose median OS was 24 months (P =.007). BCL-6 gene expression also predicted OS in an independent validation set of 39 patients with primary DLBCL (P =.01). BCL-6 protein expression, assessed by immunohistochemistry, also predicted longer OS in patients with DLBCL. BCL-6 gene expression was an independent survival predicting factor in multivariate analysis together with the elements of the International Prognostic Index (IPI) (P =.038). By contrast, the aggregate IPI score did not add further prognostic information to the patients' stratification by BCL-6 gene expression. High BCL-6 mRNA expression should be considered a new favorable prognostic factor in DLBCL and should be used in the stratification and the design of risk-adjusted therapies for patients with DLBCL. (Blood. 2001;98:945-951) PMID- 11493438 TI - Rituximab chimeric anti-CD20 monoclonal antibody treatment for adults with chronic idiopathic thrombocytopenic purpura. AB - The role of rituximab, a chimeric monoclonal antibody directed against the CD20 antigen, in the treatment of patients with chronic idiopathic thrombocytopenic purpura (ITP) has not been determined. The effectiveness and side effects of this therapeutic modality were investigated in a cohort of 25 individuals with chronic ITP. All patients had ITP that had been resistant to between 2 and 5 different therapeutic regimens, including 8 patients who had already failed splenectomy. Patients were scheduled to receive intravenous rituximab at the dose of 375 mg/m(2) once weekly for 4 weeks. Rituximab infusion-related side effects were observed in 18 patients, but were of modest intensity and did not require discontinuation of treatment. A complete response (platelet count greater than 100 x 10(9)/L) was observed in 5 cases, a partial response (platelet count between 50 and 100 x 10(9)/L) in 5 cases, and a minor response (platelet count below 50 x 10(9)/L, with no need for continued treatment) in 3 cases, with an overall response rate of 52%. In 7 cases, responses were sustained (6 months or longer). In 2 patients with relapsed disease, repeat challenge with rituximab induced a new response. In patients with a complete or partial response, a significant rise in platelet concentrations was observed early during the course of treatment, usually 1 week after the first rituximab infusion. No clinical or laboratory parameter was found to predict treatment outcome, although there was a suggestion that women and younger patients have a better chance of response. In conclusion, rituximab therapy has a limited but valuable effect in patients with chronic ITP. In view of its mild toxicity and the lack of effective alternative treatments, its use in the setting of chronic refractory ITP is warranted. (Blood. 2001;98:952-957) PMID- 11493439 TI - Thalidomide produces transfusion independence in long-standing refractory anemias of patients with myelodysplastic syndromes. AB - Thalidomide was administered to 83 patients with myelodysplastic syndrome (MDS), starting at 100 mg by mouth daily and increasing to 400 mg as tolerated. Thirty two patients stopped therapy before 12 weeks (minimum period for response evaluation), and 51 completed 12 weeks of therapy. International Working Group response criteria for MDS were used to evaluate responses. Intent-to-treat (ITT) analysis classified all off-study patients as nonresponders. Off-study patients belonged to a higher risk category (P =.002) and had a higher percentage of blasts in their pretherapy bone marrow than patients who completed 12 weeks of therapy (P =.003). No cytogenetic or complete responses were seen, but 16 patients showed hematologic improvement, with 10 previously transfusion-dependent patients becoming transfusion independent. Responders had lower pretherapy blasts (P =.016), a lower duration of pretherapy platelet transfusions (P =.013), and higher pretherapy platelets (P =.003). Among responders, 9 had refractory anemia (RA); 5 had RA with ringed sideroblasts; and 2 had RA with excess blasts. By ITT analysis, 19% of patients (16 of 83) responded, and when only evaluable patients were analyzed, 31% (16 of 51) responded. It was concluded that thalidomide, as a single agent, is effective in improving cytopenias of some MDS patients, especially those who present without excess blasts. (Blood. 2001;98:958-965) PMID- 11493440 TI - The nonexpression of CD36 on reticulocytes and mature red blood cells does not modify the clinical course of patients with sickle cell anemia. AB - It is thought that an increase in the adhesion of circulating reticulocytes to the vascular endothelium may initiate the vascular occlusion underlying the painful crises and organ failures typical of sickle cell disease (SCD). At least 2 receptors, usually present on reticulocytes, seem to be involved in this adhesion process: glycoprotein CD36 (glycoprotein IV) and integrin alpha(4)beta(1) (very late activation antigen--4). Recently, a high frequency of the platelet CD36--deficient phenotype was reported in black Africans. The frequency of this deficiency was similar in subjects with and without SCD. The role of CD36 in vaso-occlusion was then investigated by comparing the clinical course in 2 groups of black Africans homozygous for hemoglobin S, with and without CD36 deficiency, but similar in age, sex, geographical origin, number of alpha-globin genes, and beta-globin gene haplotype. Flow cytometry showed that CD36 was absent from the circulating red blood cells and reticulocytes of platelet CD36--deficient individuals but present on those from patients with normal platelet CD36 expression, and that alpha(4)beta(1) integrin levels were similar on the reticulocytes of the 2 groups. Neither clinical severity, as evaluated by the frequency and characteristics of vaso-occlusive events, nor biological data differed significantly in the 2 groups of patients. Finally, although CD36 has been suggested to play a critical role in the pathogenesis of vaso-occlusion, this study, despite including only a small number of patients, supports the idea that the modulation of expression of a single type of adhesion molecule is insufficient to counteract the pathological process leading to vaso occlusion in SCD patients. (Blood. 2001;98:966-971) PMID- 11493441 TI - Epstein-Barr virus (EBV) reactivation is a frequent event after allogeneic stem cell transplantation (SCT) and quantitatively predicts EBV-lymphoproliferative disease following T-cell--depleted SCT. AB - Reactivation of the Epstein-Barr virus (EBV) after allogeneic stem cell transplantation (allo-SCT) may evoke a protective cellular immune response or may be complicated by the development of EBV-lymphoproliferative disease (EBV-LPD). So far, very little is known about the incidence, recurrence, and sequelae of EBV reactivation following allo-SCT. EBV reactivation was retrospectively monitored in 85 EBV-seropositive recipients of a T-cell--depleted (TCD) allo-SCT and 65 EBV seropositive recipients of an unmanipulated allo-SCT. Viral reactivation (more than 50 EBV genome equivalents [gEq]/mL) was monitored frequently by quantitative real-time plasma polymerase chain reaction until day 180 after SCT. Probabilities of developing viral reactivation were high after both unmanipulated and TCD allogeneic SCT (31% +/- 6% versus 65% +/- 7%, respectively). A high CD34(+) cell number of the graft appeared as a novel significant predictor (P =.001) for EBV reactivation. Recurrent reactivation was observed more frequently in recipients of a TCD graft, and EBV-LPD occurred only after TCD-SCT. High-risk status, TCD, and use of antithymocyte globulin were predictive for developing EBV-LPD. Plasma EBV DNA quantitatively predicted EBV-LPD. The positive and negative predictive values of a viral load of 1000 gEq/mL were, respectively, 39% and 100% after TCD. Treatment-related mortality did not differ significantly between TCD and non-TCD transplants, but the incidence of chronic graft-versus-host disease was significantly less in TCD patients. It is concluded that EBV reactivation occurs frequently after TCD and unmanipulated allo-SCT, especially in recipients of grafts with high CD34(+) cell counts. EBV-LPD, however, occurred only after TCD, and EBV load quantitatively predicted EBV-LPD in recipients of a TCD graft. (Blood. 2001;98:972-978) PMID- 11493442 TI - Diagnostic utility of flow cytometric immunophenotyping in myelodysplastic syndrome. AB - The myelodysplastic syndromes (MDSs) are characterized by bilineage or trilineage dysplasia. Although diagnostic criteria are well established for MDS, a significant number of patients have blood and bone marrow findings that make diagnosis and classification difficult. Flow cytometric immunophenotyping is an accurate and highly sensitive method for detection of quantitative and qualitative abnormalities in hematopoietic cells. Flow cytometry was used to study hematopoietic cell populations in the bone marrow of 45 patients with straightforward MDS. The results were compared with those obtained in a series of patients with aplastic anemia, healthy donors, and patients with a history of nonmyeloid neoplasia in complete remission. The immunophenotypic abnormalities associated with MDS were defined, and the diagnostic utility of flow cytometry was compared, with morphologic and cytogenetic evaluations in 20 difficult cases. Although morphology and cytogenetics were adequate for diagnosis in most cases, flow cytometry could detect immunophenotypic abnormalities in cases when combined morphology and cytogenetics were nondiagnostic. It is concluded that flow cytometric immunophenotyping may help establish the diagnosis of MDS, especially when morphology and cytogenetics are indeterminate. (Blood. 2001;98:979-987) PMID- 11493443 TI - Multidrug-resistance phenotype and clinical responses to gemtuzumab ozogamicin. AB - Expression of multidrug resistance (MDR) features by acute myeloid leukemia (AML) cells predicts a poor response to many treatments. The MDR phenotype often correlates with expression of P-glycoprotein (Pgp), and Pgp antagonists such as cyclosporine (CSA) have been used as chemosensitizing agents in AML. Gemtuzumab ozogamicin, an immunoconjugate of an anti-CD33 antibody linked to calicheamicin, is effective monotherapy for CD33(+) relapsed AML. However, the contribution of Pgp to gemtuzumab ozogamicin resistance is poorly defined. In this study, blast cell samples from relapsed AML patients eligible for gemtuzumab ozogamicin clinical trials were assayed for Pgp surface expression and Pgp function using a dye efflux assay. In most cases, surface expression of Pgp correlated with Pgp function, as indicated by elevated dye efflux that was inhibited by CSA. Among samples from patients who either failed to clear marrow blasts or failed to achieve remission, 72% or 52%, respectively, exhibited CSA-sensitive dye efflux compared with 29% (P =.003) or 24% (P <.001) among samples from responders. In vitro gemtuzumab ozogamicin--induced apoptosis was also evaluated using an annexin V--based assay. Low levels of drug-induced apoptosis were associated with CSA-sensitive dye efflux, whereas higher levels correlated strongly with achievement of remission and marrow blast clearance. In vitro drug-induced apoptosis could be increased by CSA in 14 (29%) of 49 samples exhibiting low apoptosis in the absence of CSA. Together, these findings indicate that Pgp plays a role in clinical resistance to gemtuzumab ozogamicin and suggest that treatment trials combining gemtuzumab ozogamicin with MDR reversal agents are warranted. (Blood. 2001;98:988-994) PMID- 11493444 TI - The mononucleotide-dependent, nonantisense mechanism of action of phosphodiester and phosphorothioate oligonucleotides depends upon the activity of an ecto-5' nucleotidase. AB - Many reports indicate different nonantisense yet sequence-specific effects of antisense phosphorothioate oligonucleotides. Products of enzymatic degradation of the oligonucleotides can also influence cell proliferation. The cytotoxic effects of deoxyribonucleoside-5'-phosphates (dNMPs) and their 5'-phosphorothioate analogs, deoxyribonucleoside-5'-monophosphorothioates (dNMPSs) on 4 human cell types (HeLa, HL-60, K-562, and endothelial cells) were examined, and the effects were correlated with the catabolism of these compounds. The results indicate that differences in cytotoxicity of dNMPs or dNMPSs in these cells depend upon different activity of an ecto-5'-nucleotidase. It has also been found that dNMPSs stimulate proliferation of human umbilical vein endothelial cells and HL-60 cells in a concentration-dependent manner. This stimulation might be caused by the binding of deoxynucleoside-5'-phosphorothioates to as-yet unidentified nucleotide receptor(s) at the cell surface. (Blood. 2001;98:995-1002) PMID- 11493445 TI - Defective hematopoiesis and hepatic steatosis in mice with combined deficiencies of the genes encoding Fancc and Cu/Zn superoxide dismutase. AB - Several lines of evidence point to an abnormality in the response of Fanconi anemia cells to reactive oxygen species. To investigate the potential pathologic consequences of an in vivo alteration of redox state in mice lacking one of the Fanconi anemia genes, animals were generated having combined deficiencies of the cytosolic Cu/Zn superoxide dismutase (Sod1) and Fanconi anemia complementation group C (Fancc) genes. Interestingly, hepatocytes of Fancc(-/-)Sod1(-/-) mice exhibited a zonal pattern of microvesicular steatosis, possibly as a result of oxidative stress-induced injury to hepatocyte membranes. Consistent with this idea, freshly explanted Fancc(-/-)Sod1(-/-) hepatocytes demonstrated increased spontaneous production of superoxide in vitro. The second phenotypic feature of Fancc(-/-) Sod1(-/-) mice was that of bone marrow hypocellularity accompanied by significant decreases in peripheral blood erythrocyte and leukocyte numbers as compared with wild-type controls. Although flow cytometry analysis with monoclonal antibodies against cell surface antigens revealed normal numbers of primitive hematopoietic progenitor populations in Fancc(-/-)Sod1(-/-) marrow, lineage-positive progenitor numbers were significantly reduced in these mice. Furthermore, the in vitro clonogenic growth of Fancc(-/-)Sod1(-/-) erythroid, myeloid, and early B-lymphoid colonies in semisolid media was profoundly compromised. These results suggested that the altered redox state likely present in Fancc(-/-) Sod1(-/-) hematopoietic progenitors was responsible for an impairment of cell proliferation or survival. (Blood. 2001;98:1003-1011) PMID- 11493446 TI - An in vitro model of hematopoietic stem cell homing demonstrates rapid homing and maintenance of engraftable stem cells. AB - Hematopoietic stem cell (HSC) homing is believed to rely heavily on adhesion interactions between stem cells and stroma. An in vitro assay was developed for adhesion of engraftable HSCs in bone marrow suspensions to pre-established Dexter type long-term bone marrow culture stromal layers. The cell numbers in the adherent layer and supernatant were examined, along with the engraftment capability of adherent layer cells to indicate the number of HSCs that homed to in vitro stroma. The cell number in the supernatant declined over the 24-hour period. The number of test cells adhering to the stromal layer increased during the first hour and then fell at 6 and 24 hours. The number of test HSCs adhering to the stromal layer was substantial at 20 minutes, increased during the first hour, and then remained constant at 1, 6, and 24 hours of adhesion. These data indicate that adhesion of engraftable HSCs occurs quickly and increases during the first hour of contact with pre-established stroma, that adhesion plateaus within 1 hour of contact, and that HSCs maintain their engraftment capability for at least 24 hours of stromal adhesion. Long-term engraftment from test cells at more than 1 hour of adhesion represents 70.7% of the predicted engraftment from equivalent numbers of unmanipulated marrow cells, indicating that 2 of 3 test engraftable HSCs adhered. These findings demonstrate the usefulness of this model system for studying stem-stromal adhesion, allowing further dissection of the mechanism of HSC homing and exploration of possible manipulations of the process. (Blood. 2001;98:1012-1018) PMID- 11493447 TI - Thrombocytopenic c-mpl(-/-) mice can produce a normal level of platelets after administration of 5-fluorouracil: the effect of age on the response. AB - Administration of 5-fluorouracil (5-FU) to mice results in a marked increase in the level of circulating platelets in 10 days. Mice lacking Mpl, the receptor for thrombopoietin (TPO), are thrombocytopenic. To gain insight into the mechanism by which 5-FU produces such a substantial stimulation of platelet production, this study investigated whether 5-FU (150 mg/kg) produced thrombocytosis in c-mpl(-/-) mice, thus establishing whether TPO was required for this response. A 5- to 6 fold increase in platelet levels in c-mpl(-/-) mice (to approximately 1000 x 10(9)/L) was observed on days 20 and 25 after 5-FU injection. Thus, at the peak of the response, c-mpl(-/-) mice had platelet levels comparable to those in normal mice. Administration of 5-FU also produced thrombocytosis in previously splenectomized c-mpl(-/-) mice. Comparison of the platelet response to 5-FU in young (6-12 weeks) and old (33-46 weeks) c-mpl(-/-) mice found that older mice produced a much more marked response than younger mice, with a mean maximum platelet level of approximately 1700 x 10(9)/L. To determine whether this increase in circulating platelets was preceded by an increase in hematopoietic progenitors, serial cultures of bone marrow and spleen were evaluated. A considerable increase in all colony types studied was observed on days 15 and 20 in spleens of c-mpl(-/-) mice, but no similar elevations were detected in bone marrow. These results indicate that c-mpl(-/-) mice can achieve a normal level of platelets after 5-FU injection, by means of a TPO-independent mechanism, and that they respond to 5-FU myelosuppression by producing large numbers of megakaryocytic, myeloid, and erythroid progenitors. (Blood. 2001;98:1019-1027) PMID- 11493448 TI - Stromal cells expressing ephrin-B2 promote the growth and sprouting of ephrin B2(+) endothelial cells. AB - Ephrin-B2 is a transmembrane ligand that is specifically expressed on arterial endothelial cells (ECs) and surrounding cells and interacts with multiple EphB class receptors. Conversely, EphB4, a specific receptor for ephrin-B2, is expressed on venous ECs, and both ephrin-B2 and EphB4 play essential roles in vascular development. The bidirectional signals between EphB4 and ephrin-B2 are thought to be specific for the interaction between arteries and veins and to regulate cell mixing and the making of particular boundaries. However, the molecular mechanism during vasculogenesis and angiogenesis remains unclear. Manipulative functional studies were performed on these proteins in an endothelial cell system. Using in vitro stromal cells (OP9 cells) and a paraaortic splanchnopleura (P-Sp) coculture system, these studies found that the stromal cells expressing ephrin-B2 promoted vascular network formation and ephrin B2(+) EC proliferation and that they also induced the recruitment and proliferation of alpha-smooth muscle actin (alpha-SMA)-positive cells. Stromal cells expressing EphB4 inhibited vascular network formation, ephrin-B2(+) EC proliferation, and alpha-SMA(+) cell recruitment and proliferation. Thus, these data suggest that ephrin-B2 and EphB4 mediate reciprocal interactions between arterial and venous ECs and surrounding cells to form each characteristic vessel. (Blood. 2001;98:1028-1037) PMID- 11493449 TI - Platelet glycoprotein V binds to collagen and participates in platelet adhesion and aggregation. AB - Glycoprotein V (GPV) is a subunit of the platelet GPIb-V-IX receptor for von Willebrand factor and thrombin. GPV is cleaved from the platelet surface during activation by thrombin, but its role in hemostasis is still unknown. It is reported that GPV knockout mice had a decreased tendency to form arterial occluding thrombi in an intravital thrombosis model and abnormal platelet interaction with the subendothelium. In vitro, GPV-deficient platelets exhibited defective adhesion to a collagen type I-coated surface under flow or static conditions. Aggregation studies demonstrated a decreased response of the GPV deficient platelets to collagen, reflected by an increased lag phase and reduced amplitude of aggregation. Responses to adenosine diphosphate, arachidonic acid, and the thromboxane analog U46619 were normal but were enhanced to low thrombin concentrations. The defect of GPV null platelets made them more sensitive to inhibition by the anti-GPVI monoclonal antibody (mAb) JAQ1, and this was also the case in aspirin- or apyrase-treated platelets. Moreover, an mAb (V.3) against the extracellular domain of human GPV selectively inhibited collagen-induced aggregation in human or rat platelets. V.3 injected in rats as a bolus decreased the ex vivo collagen aggregation response without affecting the platelet count. Finally, surface plasmon resonance studies demonstrated binding of recombinant soluble GPV on a collagen-coupled matrix. In conclusion, GPV binds to collagen and appears to be required for normal platelet responses to this agonist. (Blood. 2001;98:1038-1046) PMID- 11493450 TI - The inflammatory action of CD40 ligand (CD154) expressed on activated human platelets is temporally limited by coexpressed CD40. AB - Recently, we have demonstrated that human platelets carry preformed CD40 ligand (CD154) molecules, which rapidly appear on the platelet surface following stimulation by thrombin. Once on the surface, platelet CD154 induces an inflammatory reaction of CD40-bearing endothelial cells. This study shows that strong platelet agonists other than thrombin also lead to the expression of CD154 on the platelet surface. At the same time, several lines of evidence are presented that together indicate that thrombotic events in the vasculature are generally accompanied by activation of the inflammatory potential of platelet CD154. This study also reports the constitutive expression of CD40, the receptor for CD154, on platelets. The binding of CD154 to coexpressed CD40 in the platelet aggregate leads within minutes to hours to the cleavage of membrane-bound surface CD154 and the release of an 18-kd soluble form of the molecule. Soluble CD154 (sCD154), in contrast to transmembrane CD154, can no longer induce an inflammatory reaction of endothelial cells. These findings indicate that the interaction of platelet CD154 with CD40 on neighboring cells is temporally limited to prevent an uncontrolled inflammation at the site of thrombus formation. Thus, similar to the very tight regulation of the CD154-CD40 interaction in the immune system, an effective mechanism controls the inflammatory potential of platelet CD154 in the vascular system. (Blood. 2001;98:1047-1054) PMID- 11493451 TI - Variable protection of beta 3-integrin--deficient mice from thrombosis initiated by different mechanisms. AB - Platelet integrin alpha IIb beta 3 (GPIIb/IIIa) plays a central role in the initiation of arterial thrombosis, but its contribution to disseminated microvascular thrombosis is less well defined. Therefore, wild-type mice (beta 3(+/+)), beta 3-integrin-deficient mice (beta 3(-/-)), and wild-type mice treated with a hamster monoclonal antibody (1B5) that blocks murine alpha IIb beta 3 function were tested in models of large-vessel and microvascular thrombosis. In the large-vessel model, ferric chloride was used to injure the carotid artery, and the time to thrombosis was measured. In beta 3(+/+) mice, the median time to occlusion was 6.7 minutes, whereas occlusion did not occur in any of the beta 3( /-) mice tested (P <.001). Fab and F(ab')(2) fragments of 1B5 increased the median time to occlusion. To initiate systemic intravascular thrombosis, prothrombotic agents were administered intravenously, and platelet thrombus formation was monitored by the decrease in circulating platelet count. Three minutes after the injection of adenosine diphosphate (ADP), collagen + epinephrine, or tissue factor, the platelet counts in beta 3(+/+) mice decreased by 289, 424, and 429 x 10(3)/microL, respectively. beta 3(-/-) mice and wild-type mice pretreated with 1B5 Fab (1 mg/kg, IP) were nearly completely protected from the effects of ADP. In contrast, beta 3(-/-) mice were only partially protected from the effects of collagen + epinephrine and minimally protected from the effects of tissue factor. In all cases, less fibrin became deposited in the lungs of beta 3(-/-) mice than in wild-type mice. These results suggest that though alpha IIb beta 3 plays a dominant role in large-vessel thrombosis, it plays a variable role in systemic intravascular thrombosis. (Blood. 2001;98:1055-1062) PMID- 11493452 TI - Identification of a region in glycoprotein IIIa involved in subunit association with glycoprotein IIb: further lessons from Iraqi-Jewish Glanzmann thrombasthenia. AB - The most frequent mutation causing Glanzmann thrombasthenia in Iraqi-Jews (IJ-1) is an 11-bp deletion in exon 13 of the glycoprotein (GP) IIIa gene. This deletion predicts a frameshift that results in the elimination of the C406-C655 disulfide bond and a premature termination codon shortly before the transmembrane domain. To determine the contribution of each of these alterations to the thrombasthenic phenotype, Chinese hamster ovary or baby hamster kidney cells were cotransfected with normal GPIIb complementary DNA (cDNA) and the following GPIIIa cDNAs: normal, cDNA bearing IJ-1 mutation, 2011T>A mutated cDNA predicting C655S (single letter amino acid codes) substitution, and 2019A>T mutated cDNA predicting Stop657. Elimination of the C406-C655 disulfide bond by C655S substitution did not affect GPIIb/IIIa surface expression or binding of the transfected cells to immobilized fibrinogen, whereas elimination of the transmembrane and cytoplasmic domains in IJ-1 and Stop657 mutants prevented both surface expression and binding of the transfected cells to immobilized fibrinogen. Immunohistochemical staining and immunoprecipitation demonstrated that the elimination of amino acids 657-762 in IJ-1 and Stop657 prevented intracellular GPIIb/IIIa complex formation, and differential immunofluorescence staining of GPIIIa and cellular organelles suggested that the truncated uncomplexed GPIIIa protein was retained in the endoplasmic reticulum. Because the use of GPIIIa Stop693 and normal GPIIb cDNAs yielded GPIIb/IIIa complex formation, though with lower efficiency, it is suggested that amino acids 657-692 of GPIIIa are essential for the intracellular association of GPIIb and GPIIIa. (Blood. 2001;98:1063-1069) PMID- 11493453 TI - Cross-presentation of tumor antigens by bone marrow-derived antigen-presenting cells is the dominant mechanism in the induction of T-cell tolerance during B cell lymphoma progression. AB - Tumor antigen-specific T-cell tolerance may limit the efficacy of therapeutic cancer vaccines. Direct presentation of antigens by tumor cells incapable of providing adequate costimulation to tumor-specific T cells has been suggested as the basis for this unresponsiveness. Using parent-into-F1 bone marrow (BM) chimeras, this study unambiguously demonstrates that the induction of this tolerant state requires T-cell recognition of tumor antigen presented by BM derived antigen-presenting cells (APCs), not tumor cells themselves. In the absence of host APC presentation, tumor-specific T cells remained functional, even in the setting of antigen expressed by B-cell lymphomas residing in secondary lymphoid tissues. The intrinsic APC capacity of tumor cells has therefore little influence over T-cell priming versus tolerance, a decision that is regulated at the level of host APCs. (Blood. 2001;98:1070-1077) PMID- 11493454 TI - Antiapoptotic mechanism of HIV protease inhibitors: preventing mitochondrial transmembrane potential loss. AB - Treatment of cells with the HIV drugs ritonavir, saquinavir, or nelfinavir (Nfv) inhibits apoptosis induced by a variety of stimuli. Because these drugs are protease inhibitors, they have been postulated to inhibit apoptosis by blocking caspase activity. This study shows that Nfv has no effect on caspase activity or on the transcription or synthesis of a variety of apoptosis regulatory molecules. Instead, Nfv inhibits mitochondrial transmembrane potential loss (Delta psi(m)) and the subsequent release of apoptotic mediators. Consequently, the antiapoptotic effects of Nfv are restricted to apoptotic pathways that involve Delta psi(m). (Blood. 2001;98:1078-1085) PMID- 11493455 TI - Cdc42, Rac1, and the Wiskott-Aldrich syndrome protein are involved in the cytoskeletal regulation of B lymphocytes. AB - Patients with the immunodeficiency disorder Wiskott-Aldrich syndrome (WAS) have lymphocytes with aberrant microvilli, and their T cells, macrophages, and dendritic cells are impaired in cytoskeletal-dependent processes. WAS is caused by a defective or a missing WAS protein (WASP). Signal mediators interleukin-4 (IL-4) and CD40 are important for actin-dependent morphology changes in B cells. A possible function of WASP and its interacting partners, Cdc42 and Rac1, was investigated for these changes. It was found that active Cdc42 and Rac1 induced filopodia and lamellipodia, respectively, in activated B cells. Evidence is given that IL-4 has a specific role in the regulated cycling of Cdc42 because IL-4 partially and transiently depleted active Cdc42 from detergent extract of activated B cells. WASP-deficient B lymphocytes were impaired in IL-4-- and CD40 dependent induction of polarized and spread cells. Microvilli were expressed on WASP-deficient B cells, but they appeared shorter and less dense in cell contacts than in wild-type cells. In conclusion, evidence is provided for the involvement of Cdc42, Rac1, and WASP in the cytoskeletal regulation of B lymphocytes. Aberrations in WASP-deficient B lymphocytes, described here, provide further evidence that WAS is a cytoskeletal disease of hematopoietic cells. (Blood. 2001;98:1086-1094) PMID- 11493456 TI - Therapeutic efficacy of intravenous immunoglobulin preparations depends on the immunoglobulin G dimers: studies in experimental immune thrombocytopenia. AB - The clinical benefit of intravenous immunoglobulin (IVIG) preparations in the treatment of immune thrombocytopenic purpura (ITP) is supposed to be mediated by blockade of Fc gamma receptor--bearing phagocytes. In 2 experimental models for ITP, it is shown that the therapeutic efficacy of IVIG preparations is related to the IgG dimer content present in these preparations. A rat monoclonal antibody (mAb; MWReg30) directed to the murine platelet-specific integrin alpha(IIb)beta(3) (gpIIb/IIIa) was administered intraperitoneally either as bolus injection or continuous infusion. With bolus injection, the circulating platelet count dropped to almost zero within 3 hours. Pretreatment with cobra venom factor did not affect platelet depletion, whereas pretreatment with anti-Fc gamma RII/III mAb 2.4G2 or IVIG greatly reduced platelet clearance. With continuous infusion, platelet numbers reached a steady state after 4 days, at approximately 25% of control. This reduction in platelets was, however, not observed in mice deficient for the FcR gamma-chain, lacking Fc gamma RI, Fc gamma RIII, and Fc gamma RIII(-/-) mice. Infusion of a single dose of IVIG with a high IgG dimer content on the 4th day--ie, mimicking therapeutic administration--resulted in a platelet increase for several days. IVIG predominantly consisting of monomeric IgG had no effect on platelet numbers. In conclusion, continuous infusion of MWReg30 induces thrombocytopenia in mice by enhancing Fc gamma receptor--mediated clearance of platelets. In this model, it is shown that IgG dimers present in IVIG preparations are responsible for the increase in platelet counts. (Blood. 2001;98:1095-1099) PMID- 11493457 TI - Age-related increase of frequency of a new, phenotypically distinct subpopulation of human peripheral blood T cells expressing lowered levels of CD4. AB - Aging is associated with modifications of T-cell phenotype and function, leading to impaired activation in response to both new and recall antigens. It is not known if T-cell activation results in elimination of a number of the CD4 molecules from the cell surface, as is the case with CD3/T-cell receptor complexes, or how aging influences the process. The T cells of young and elderly donors with reduced expression of CD4 were examined to see whether these cells exhibit other phenotypic features suggesting their active state. It was found that T lymphocytes expressing CD4 can be divided into 2 semidiscrete subpopulations: the major (CD4(+)) population, in which the level of expression of CD4 is constant and high, and a minor population (CD4(lo)), in which the expression of CD4 can be up to an order of magnitude lower than on the CD4(+) cells. The proportion of CD4(lo) cells is age dependent and highly variable in the apparently healthy human population, with the expression of CD4 ranging from around 10% of all peripheral blood lymphocytes in the young to more than 30% in the elderly. Lowered expression of CD4 is correlated with a reduced expression of CD3, as well as with a decreased amount of CD28 and CD95Fas. Activation of CD4(lo) cells is suggested by their expression of CD25 and increased amounts of HLA-DR. Phenotypic characteristics of the CD4(lo) T-cell subpopulation suggest that it might be formed by (perhaps chronically) activated, temporarily apoptosis resistant cells, possibly accumulating in the elderly. (Blood. 2001;98:1100-1107) PMID- 11493458 TI - Tumor necrosis factor-alpha induces coordinated changes in major histocompatibility class I presentation pathway, resulting in increased stability of class I complexes at the cell surface. AB - It is demonstrated that similar to interferon gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) induces coordinated changes at different steps of the major histocompatibility complex (MHC) class I processing and presentation pathway in nonprofessional antigen-presenting cells (APCs). TNF-alpha up regulates the expression of 3 catalytic immunoproteasome subunits--LMP2, LMP7, and MECL-1--the immunomodulatory proteasome activator PA28 alpha, the TAP1/TAP2 heterodimer, and the total pool of MHC class I heavy chain. It was also found that in TNF-alpha--treated cells, MHC class I molecules reconstitute more rapidly and have an increased average half-life at the cell surface. Biochemical changes induced by TNF-alpha in the MHC class I pathway were translated into increased sensitivity of TNF-alpha--treated targets to lysis by CD8(+) cytotoxic T cells, demonstrating improved presentation of at least certain endogenously processed MHC class I--restricted peptide epitopes. Significantly, it was demonstrated that the effects of TNF-alpha observed in this experimental system were not mediated through the induction of IFN-gamma. It appears to be likely that TNF-alpha- mediated effects on MHC class I processing and presentation do not involve any intermediate messengers. Collectively, these data demonstrate the existence of yet another biologic activity exerted by TNF-alpha, namely its capacity to act as a coordinated multi-step modulator of the MHC class I pathway of antigen processing and presentation. These results suggest that TNF-alpha may be useful when a concerted up-regulation of the MHC class I presentation machinery is required but cannot be achieved by IFN-gamma. (Blood. 2001;98:1108-1115) PMID- 11493459 TI - Quantitation of T-cell neogenesis in vivo after allogeneic bone marrow transplantation in adults. AB - Following myeloablative therapy, it is unknown to what extent age-dependent thymic involution limits the generation of new T cells with a diverse repertoire. Normal T-cell receptor gene rearrangement in T-cell progenitors results in the generation of T-cell receptor rearrangement excision circles (TRECs). In this study, a quantitative assay for TRECs was used to measure T-cell neogenesis in adult patients with leukemia who received myeloablative therapy followed by transplantation of allogeneic hematopoietic stem cells. Although phenotypically mature T cells had recovered by 1 to 2 months after bone marrow transplantation (BMT), TREC levels remained low for 3 months after BMT. T-cell neogenesis became evident by 6 months, and normal levels of adult thymic function were restored at 6 to 12 months after BMT. Subsequent leukemia relapse in some patients was associated with reduced TREC levels, but infusion of mature donor CD4(+) T cells resulted in rapid restoration of thymic function. These studies demonstrate that T-cell neogenesis contributes to immune reconstitution in adult patients and suggest that thymic function can be manipulated in vivo. (Blood. 2001;98:1116 1121) PMID- 11493460 TI - Peripheral blood stem cell allograft rejection mediated by CD4(+) T lymphocytes recognizing a single mismatch at HLA-DP beta 1*0901. AB - Little is known about the molecular characteristics of alloantigens recognized by alloreactive T cells mediating hematologic stem cell graft rejection. In particular, it has never been shown that such alloantigens can be encoded by HLA DP beta alleles. Indeed, matching for HLA-DP antigens is generally not considered to be of functional importance for the outcome of allogeneic bone marrow or peripheral blood stem cell transplantation. In this study, a case of peripheral blood stem cell allograft rejection was investigated in which the patient and donor differed for a single mismatch at HLA-DP in the rejection direction. Patient-derived T lymphocytes circulating at the time of rejection showed direct ex vivo cytotoxic activity against donor-derived B-lymphoblastoid cells as well as other HLA-DP beta 1*0901--expressing targets. The presence of HLA-DP beta 1*0901--specific effectors in vivo was further confirmed by in vitro stimulation experiments. CD4(+) T-cell lines and clones with specific cytotoxic activity against HLA-DP beta 1*0901--expressing targets including donor B-lymphoblastoid cells were generated both by nonspecific and by donor-specific in vitro stimulation. Taken together, these data demonstrate that HLA-DP can be the target antigen of cytotoxic CD4(+) T lymphocytes involved in peripheral blood stem cell allograft rejection. (Blood. 2001;98:1122-1126) PMID- 11493461 TI - Gene expression screening of human mast cells and eosinophils using high-density oligonucleotide probe arrays: abundant expression of major basic protein in mast cells. AB - Mast cells (MCs) and eosinophils are thought to play important roles in evoking allergic inflammation. Cell-type--specific gene expression was screened among 12,000 genes in human MCs and eosinophils with the use of high-density oligonucleotide probe arrays. In comparison with other leukocytes, MCs expressed 140 cell-type--specific transcripts, whereas eosinophils expressed only 34. Among the transcripts for expected MC-specific proteins such as tryptase, major basic protein (MBP), which had been thought to be eosinophil specific, was ranked fourth in terms of amounts of increased MC-specific messenger RNA. Mature eosinophils were almost lacking this transcript. MCs obtained from 4 different sources (ie, lung, skin, adult peripheral blood progenitor--derived and cord blood progenitor--derived MCs, and eosinophils) were found to have high protein levels of MBP in their granules with the use of flow cytometric and confocal laser scanning microscopic analyses. The present finding that MCs can produce abundant MBP is crucial because many reports regarding allergic pathogenesis have been based on earlier findings that MBP was almost unique to eosinophils and not produced by MCs. (Blood. 2001;98:1127-1134) PMID- 11493462 TI - Th2 polarization by Der p 1--pulsed monocyte-derived dendritic cells is due to the allergic status of the donors. AB - The polarization of the immune response toward a Th2 or a Th1 profile can be mediated by dendritic cells (DCs) following antigen presentation and interaction with T cells. Costimulatory molecules such as CD80 and CD86 expressed by DCs, the polarizing cytokine environment during DC--T-cell interaction, and also the nature of the antigen are critical in the orientation of the immune response. In this study, the effect of the cysteine protease Der p 1, one of the major allergens of the house dust mite Dermatophagoides pteronyssinus, on these different parameters was evaluated comparatively on monocyte-derived DCs obtained from healthy donors, from pollen-sensitive patients, or from patients sensitive to Dermatophagoides pteronyssinus. Results showed that Der p 1 induced an increase in CD86 expression only on DCs from house dust mite--sensitive patients. This was also associated with a higher capacity to induce T-cell proliferation, a rapid increase in the production of proinflammatory cytokines, tumor necrosis factor--alpha and interleukin (IL)-1 beta, and the type 2 cytokine IL-10. No changes in the release of IL-12 p70 were induced by Der p 1. Finally, purified T cells from house dust mite-sensitive patients stimulated by autologous Der p 1- pulsed DCs preferentially produced IL-4 rather than interferon-gamma. These effects were abolished in the presence of the inactive precursor of Der p 1 (ProDer p 1). Taken together, these data suggest that DCs from house dust mite- sensitive patients, in contrast to DCs from healthy donors and from pollen sensitive patients, exposed to Der p 1 play a pivotal role in the enhancement of the Th2 response associated with the allergic reaction developed in response to house dust mite exposure. (Blood. 2001;98:1135-1141) PMID- 11493463 TI - Configuration of human dendritic cell cytoskeleton by Rho GTPases, the WAS protein, and differentiation. AB - The cellular mechanisms that configure the cytoskeleton during migration of dendritic cells (DCs) are poorly understood. Immature DCs assemble specialized adhesion structures known as podosomes at their leading edge; these are associated with the localized recruitment of the Wiskott-Aldrich Syndrome protein (WASp) and the actin organizing actin-related protein 2/3 complex. In immature DCs lacking WASp, podosomes are absent, residual dysmorphic lamellipodia and filopodia are nonpolarized, and migration is severely compromised. Microinjection studies indicate that podosome assembly and polarization require concerted action of Cdc42, Rac, and Rho, thereby providing a link between sequential protrusive and adhesive activity. Formation of podosomes is restricted to cells with an immature phenotype, indicating a specific role for these structures during the early migratory phase. (Blood. 2001;98:1142-1149) PMID- 11493464 TI - Autocrine antiapoptotic stimulation of cultured adult T-cell leukemia cells by overexpression of the chemokine I-309. AB - Adult T-cell leukemia (ATL) is an aggressive malignancy of CD4(+) T cells caused by the human T-cell leukemia virus type 1 (HTLV-1). The viral leukemogenesis is critically dependent on its oncoprotein Tax because the protein as well as the virus can immortalize primary human lymphocytes to permanent growth. As a transcriptional transactivator, Tax can stimulate the expression of distinct cellular genes. Alterations in the expression levels of unknown growth-relevant genes may contribute to the changed growth properties of Tax-immortalized and leukemic cells. To identify genes that are linked to Tax transformation and ATL leukemogenesis, this study systematically compared the gene expression of cultured cells from patients with acute ATL with that of stimulated peripheral blood T lymphocytes. Several overexpressed RNAs that encode signal transduction functions were identified. These include a dual-specific protein phosphatase (PAC1), an interferon-inducible factor (ISG15), a basic helix-loop-helix transcription factor (DEC-1), and the secreted antiapoptotic chemokine I-309. The ATL cell culture supernatants contained an antiapoptotic activity that could be specifically inhibited by antibodies directed against I-309. Inhibition of I-309 receptor (CCR8) signaling by pertussis toxin increased the apoptosis rate of ATL cell cultures in the presence and absence of external apoptotic stimuli. Both the I-309--specific antiapoptotic activity and the proapoptotic effect of inhibitors of I-309 signaling suggest the existence of an antiapoptotic autocrine loop in ATL cells. Thus, the overexpression of this chemokine may inhibit apoptosis in ATL cells and could substantially contribute to their growth. (Blood. 2001;98:1150-1159) PMID- 11493465 TI - Expression of functional lung resistance--related protein predicts poor outcome in adult T-cell leukemia. AB - Chemotherapy of patients with adult T-cell leukemia (ATL) has been unsuccessful. The poor outcome is thought to be caused mainly by the drug resistance of ATL cells. Lung resistance-related protein (LRP) is a novel protein associated with drug resistance. The expression of LRP messenger RNA (mRNA) was evaluated by slot blot analysis in 55 patients with ATL. Of these patients, 36 had acute, 12 chronic, and 7 lymphoma-type ATL. The expression levels of LRP mRNA were significantly higher in chronic ATL than in lymphoma-type ATL (P =.007). The expression of LRP mRNA was higher in patients with white blood cell counts above 30,000/microL (P =.038) or with abnormal lymphocyte counts above 10,000/microL (P =.007) than in the remaining patients. The enhanced efflux of [(14)C]doxorubicin from nuclei isolated from ATL cells that expressed high levels of LRP was inhibited by a polyclonal antibody against LRP, and the accumulation of doxorubicin in the isolated nuclei was increased by the anti-LRP antibody. In acute and lymphoma-type ATL patients, high expression of LRP mRNA at diagnosis correlated with shorter survival, and a Cox proportional hazards model showed that LRP expression is an independent prognostic factor. These findings suggest that functionally active LRP is expressed in some ATL cells and that it is involved in drug resistance and poor prognosis in ATL. (Blood. 2001;98:1160-1165) PMID- 11493466 TI - A leukemic stem cell with intrinsic drug efflux capacity in acute myeloid leukemia. AB - The hematopoietic stem cell underlying acute myeloid leukemia (AML) is controversial. Flow cytometry and the DNA-binding dye Hoechst 33342 were previously used to identify a distinct subset of murine hematopoietic stem cells, termed the side population (SP), which rapidly expels Hoechst dye and can reconstitute the bone marrow of lethally irradiated mice. Here, the prevalence and pathogenic role of SP cells in human AML were investigated. Such cells were found in the bone marrow of more than 80% of 61 patients and had a predominant CD34(low/-) immunophenotype. Importantly, they carried cytogenetic markers of AML in all 11 cases of active disease examined and in 2 out of 5 cases in complete hematological remission. Comparison of daunorubicin and mitoxantrone fluorescence emission profiles revealed significantly higher drug efflux from leukemic SP cells than from non-SP cells. Three of 28 SP cell transplants generated overt AML like disease in nonobese diabetic--severe combined immunodeficient mice. Low but persistent numbers of leukemic SP cells were detected by molecular and immunological assays in half of the remaining mice. Taken together, these findings indicate that SP cells are frequently involved in human AML and may be a target for leukemic transformation. They also suggest a mechanism by which SP cells could escape the effects of cytostatic drugs and might eventually contribute to leukemia relapse. (Blood. 2001;98:1166-1173) PMID- 11493467 TI - Tumor cells of hairy cell leukemia express multiple clonally related immunoglobulin isotypes via RNA splicing. AB - Hairy cell leukemia (HCL) derives from a mature B cell and expresses markers associated with activation. Analysis of immunoglobulin variable region genes has revealed somatic mutation in most cases, consistent with an origin from a cell that has encountered the germinal center. One unusual feature of hairy cells (HCs) is the frequent expression of multiple immunoglobulin heavy chain isotypes, with dominance of immunoglobulin (Ig)--G3, but only a single light chain type. The origin and clonal relationship of these isotype variants have been unclear. In order to probe the isotype switching status of HCL, RNA transcripts of V(H)DJ(H)--constant region sequences from 5 cases of typical HCL, all expressing multiple surface immunoglobulin isotypes, were analyzed. Tumor V(H)DJ(H)--C(mu) sequences were identified and found to be somatically mutated (range, 1.4% to 6.5%), with a low level of intraclonal heterogeneity. Additional immunoglobulin isotypes of identical V(H)DJ(H) sequence were also identified, including IgD (5 of 5), IgG3 (5 of 5), IgG1 (3 of 5), IgG2 (2 of 5), IgA1 (4 of 5), and IgA2 (1 of 5). Derivation of multiple isotypes from individual cells was demonstrated by analyzing transcripts in single sorted cells from one patient, with evidence for coexistence of isotype variants in 10 of 10 cells. These findings indicate that clonally related multiple isotypes coexist in single HCs, with individual isotypes presumably generated via RNA splicing. Production of IgG3 appears common, but IgG1, IgG2, IgA1, and IgA2 also arise, indicating a continuing influence of a directed process on the tumor clone. These HCs appear to be arrested at the point of isotype switch, where RNA processing may precede deletional recombination. (Blood. 2001;98:1174-1181) PMID- 11493468 TI - T(11;18)(q21;q21) is associated with advanced mucosa-associated lymphoid tissue lymphoma that expresses nuclear BCL10. AB - The development of gastric mucosa-associated lymphoid tissue (MALT) lymphoma is a multistep process and can be clinico-pathologically divided into Helicobacter pylori-associated gastritis, low-grade tumors, and high-grade tumors. The molecular events underlying this progression are largely unknown. However, identification of the genes involved in MALT lymphoma-specific t(11;18)(q21;q21) and t(1;14)(p22;q32) has provided fresh insights into the pathogenesis of this disease. T(11;18)(q21;q21) results in a chimeric transcript between the API2 and the MALT1 genes, whereas t(1;14) (p22;q32) causes aberrant nuclear BCL10 expression. Significantly, nuclear BCL10 expression also occurs frequently in MALT lymphomas without t(1;14)(p22;q32), suggesting an important role for BCL10 in lymphoma development. Thirty-three cases of H pylori gastritis, 72 MALT lymphomas, and 11 mucosal diffuse large B-cell lymphomas (DLBCL) were screened for t(11;18)(q21;q21) by reverse transcription-polymerase chain reaction followed by sequencing. BCL10 expression in lymphoma cases was examined by immunohistochemistry. The API2--MALT1 fusion transcript was not detected in H pylori gastritis and mucosal DLBCL but was found in 25 of 72 (35%) MALT lymphomas of various sites. Nuclear BCL10 expression was seen in 28 of 53 (53%) of MALT lymphomas. Of the gastric cases, the largest group studied, the frequency of both t(11;18)(q21;q21) and nuclear BCL10 expression was significantly higher in tumors that showed dissemination to local lymph nodes or distal sites (14 of 18 = 78% and 14 of 15 = 93%, respectively) than those confined to the stomach (3 of 29 = 10% and 10 of 26 = 38%). Furthermore, t(11;18)(q21;q21) closely correlated with BCL10 nuclear expression. These results indicate that both t(11;18)(q21;q21) and BCL10 nuclear expression are associated with advanced MALT lymphoma and that their oncogenic activities may be related to each other. (Blood. 2001;98:1182 1187) PMID- 11493469 TI - Loss of heterozygosity in childhood de novo acute myelogenous leukemia. AB - A genome-wide screening for loss of heterozygosity (LOH), a marker for possible involvement of tumor suppressor genes, was conducted in 53 children with de novo acute myelogenous leukemia (AML). A total of 177 highly polymorphic microsatellite repeat markers were used in locus-specific polymerase chain reactions. This comprehensive allelotyping employed flow-sorted cells from diagnostic samples and whole-genome amplification of DNA from small, highly purified samples. Nineteen regions of allelic loss in 17 patients (32%) were detected on chromosome arms 1q, 3q, 5q, 7q (n = 2), 9q (n = 4), 11p (n = 2), 12p (n = 3), 13q (n = 2), 16q, 19q, and Y. The study revealed a degree of allelic loss underestimated by routine cytogenetic analysis, which failed to detect 9 of these LOH events. There was no evidence of LOH by intragenic markers for p53, Nf1, or CBFA2/AML1. Most lymphocytes lacked the deletions, which were detected only in the leukemic myeloid blast population. Analysis of patients' clinical and biologic characteristics indicated that the presence of LOH was associated with a white blood cell count of 20 x 10(9)/L or higher but was not correlated with a shorter overall survival. The relatively low rate of LOH observed in this study compared with findings in solid tumors and in pediatric acute lymphoblastic leukemia and adult AML suggests that tumor suppressor genes are either infrequently involved in the development of pediatric de novo AML or are inactivated by such means as methylation and point mutations. Additional study is needed to determine whether these regions of LOH harbor tumor suppressor genes and whether specific regions of LOH correlate with clinical characteristics. (Blood. 2001;98:1188-1194) PMID- 11493470 TI - The Kit-activating mutation D816V enhances stem cell factor--dependent chemotaxis. AB - The D816V mutation of c-kit has been detected in patients with mastocytosis. This mutation leads to constitutive tyrosine kinase activation of Kit. Because stem cell factor (SCF), the ligand for Kit (CD117(+)), is a chemoattractant for CD117(+) cells and one feature of mastocytosis is an abnormal collection of mast cells in tissues derived from CD34(+)CD117(+) mast cell precursors, the hypothesis was considered that the D816V mutation would enhance chemotaxis of these precursor cells. Constructs encoding wild-type Kit or Kit bearing the D816V mutation were transfected into Jurkat cells, labeled with Calcein-AM, and migration to SCF assessed in the presence or absence of tyrosine kinase inhibitors. Chemotaxis to SCF was enhanced in D816V transfectants compared to wild-type Kit transfectants (P <.002). Migration of both transfectants was inhibited by tyrosine kinase inhibitors, although D816V transfectants were more sensitive. Chemotaxis was next performed on CD34(+)CD117(+) circulating mast cell precursors obtained from patients with mastocytosis. Analysis of prechemotaxis and migrated cells showed that whereas less than 10% in the prechemotaxis sample had the D816V mutation, 40% to 80% of migrated cells had this mutation. These results demonstrate that the D816V Kit mutation enhances chemotaxis of CD117(+) cells, offering one explanation for increased mast cells observed in tissues of patients with mastocytosis. (Blood. 2001;98:1195-1199) PMID- 11493471 TI - The contribution of NF-kappa B activity to spontaneous proliferation and resistance to apoptosis in human T-cell leukemia virus type 1 Tax-induced tumors. AB - Human T-cell leukemia virus type I is the etiologic agent of adult T-cell leukemia/lymphoma. The Tax protein of this virus is thought to contribute to cellular transformation and tumor development. In this report, we have used a Tax transgenic mouse model of tumorigenesis to study the contribution of nuclear factor (NF)-kappa B activity to spontaneous tumor cell proliferation and resistance to apoptosis. We have demonstrated elevated expression levels of NF kappa B--inducible cytokines, including interleukin (IL)-6, IL-10, IL-15, and interferon (IFN)-gamma, in freshly isolated primary tumors from Tax transgenic mice. Inhibitors of NF-kappa B activity, sodium salicylate and cyclopentenone prostaglandins (prostaglandin A(1) and 15-deoxy-Delta(12,14)-prostaglandin J(2)), blocked spontaneous proliferation of Tax transgenic mouse spleen cells. In addition, Tax-induced tumor cells, which are resistant to irradiation-induced apoptosis, became sensitive to apoptosis in the presence of sodium salicylate and prostaglandins. These results strongly suggest that Tax-mediated induction of NF kappa B activity contributes to tumorigenesis in vivo. (Blood. 2001;98:1200-1208) PMID- 11493472 TI - Unusual childhood extramedullary hematologic malignancy with natural killer cell properties that contains tropomyosin 4--anaplastic lymphoma kinase gene fusion. AB - This report describes an unusual extramedullary hematologic malignancy in an 18 month-old child who presented with a capillary leak syndrome that evolved into hyperleukocytosis with malignant cells. The circulating tumor cells did not express an antigen profile typical of any subtype of leukemia commonly observed in children. Tumor cells were CD3(-)/CD56(+); had germline TCR genes; and strongly expressed CD30, epithelial membrane antigen, and anaplastic lymphoma kinase (ALK) consistent with a null cell anaplastic large cell lymphoma (ALCL). The malignant cells contained a t(2;19)(p23;p13.1) that interrupted ALK and translocated it to the der(19). Reverse transcriptase-polymerase chain reaction and nucleotide sequence analysis revealed fusion of ALK to tropomyosin 4, an ALK fusion partner not described previously in hematologic malignancies. The clinical presentation and phenotypic features of this malignancy were not typical for ALCL because tumor cells expressed both myeloid (CD13, CD33, HLA-DR) and natural killer (NK) cell antigens. The neoplastic cells most resembled NK cells because in addition to being CD3(-)/CD56(+) with germline TCR genes, these cells were CD25(+)/CD122(+)/granzyme B(+) and possessed the functional properties of immature NK cells. The unusual clinical presentation, immunophenotype, and functional properties of these neoplastic cells suggest that this malignancy may be derived from the putative myeloid-NK precursor cell. Furthermore co-expression of NK and ALCL features supports the concept that a minority of null-ALCL may be derived from NK cells and expands the spectrum of phenotypes that can be seen in tumors produced by ALK fusion proteins. (Blood. 2001;98:1209-1216) PMID- 11493473 TI - Synthetic unmethylated cytosine-phosphate-guanosine oligodeoxynucleotides are potent stimulators of antileukemia responses in naive and bone marrow transplant recipients. AB - Immunostimulatory cytosine-phophate-guanosine (CpG)--containing motifs in bacterial DNA are potent immune system activators. Depending on the bases flanking the CpG motif and on the DNA backbone, CpG oligodeoxynucleotides (ODNs) can induce relatively more B-cell activation or relatively more natural killer (NK)--cell activation. To evaluate their antitumor activities, an NK-optimized ODN (1585) and 2 B-cell--optimized ODNs (1826 and 2006) were compared for their ability to protect naive mice against a lethal acute myelogenous leukemia (AML) challenge. CpG 2006, but not CpG 1585, administered 2 days before the AML challenge, allowed mice to survive more than 100 times a lethal tumor dose. Cell depletion studies showed that protection did not require T or B cells but depended on NK cells and also on an NK-independent mechanism. CpG 2006 protected against AML challenge in both syngeneic and allogeneic bone marrow transplant (BMT) recipients at both early and late time points after transplantation. Although CpG 1585 had no protective effect on its own, it showed a striking synergy with CpG 2006 to induce prolonged survival to AML challenge in allogeneic recipients of T-cell-depleted marrow grafts, exceeding the survival benefit of donor lymphocyte infusion (DLI). When combined with DLI, a synergistic effect was observed in recipients of CpG2006 or 2006 + 1585 with 88% of mice surviving long term. These data are the first to indicate that the systemic administration of CpG ODNs is a potent means of inducing therapeutic anti-AML innate immune responses in naive and BMT recipients. (Blood. 2001;98:1217-1225) PMID- 11493474 TI - Role of the liver in regulating numbers of circulating neutrophils. AB - Neutrophils (polymorphonuclear leukocytes [PMNs]) carry potent destructive enzymes that can destroy invasive bacteria or damage normal tissue. PMNs have a half-life of only 6 hours in the blood, but the details of this homeostasis are unknown. In a rat model of endotoxemia, P-selectin was selectively up-regulated in hepatic sinusoids and veins where it was necessary for phagocytosis of PMNs by Kupffer cells in the liver, as opposed to the spleen or the lungs. Apoptotic PMNs appeared in the lungs and spleen only after inactivation of Kupffer cells by gadolinium chloride (GdCl(3)). Blocking of Fas protein reduced the number of apoptotic cells in the liver; binding of annexin V to phosphatidylserine (PS) reduced the number of PMNs phagocytosed by Kupffer cells. The results support a clearance pathway in which apoptosis and phagocytosis are effected by Kupffer cells after P-selectin-mediated sequestration. (Blood. 2001;98:1226-1230) PMID- 11493475 TI - Molecular basis of biomaterial-mediated foreign body reactions. AB - Despite being inert and nontoxic, implanted biomaterials often trigger adverse foreign body reactions such as inflammation, fibrosis, infection, and thrombosis. With regard to the inflammatory responses to biomaterial implants, it was previously found that a crucial precedent event was the spontaneous adsorption and denaturation of fibrinogen on implant surfaces. It was further found that interactions between the phagocyte integrin Mac-1 (CD11b/CD18) and one short sequence within the fibrinogen D domain (gamma 190-202; P1) at least partially explained phagocyte accumulation on implant surfaces. However, the reason that adsorbed fibrinogen is proinflammatory--while soluble fibrinogen clearly is not- remained obscure. In this study, therefore, the question of how fibrinogen is converted to a proinflammatory state when adsorbed to biomaterial surfaces is investigated. In soluble fibrinogen, the 13 amino acid P1 sequence was found to be hidden. However, the adsorption and denaturation of fibrinogen on the surfaces of commonly used biomaterials lead to the exposure of P1 and a second neo epitope, gamma 377-395 (P2), which also interacts with Mac-1 and is similarly occult in the soluble protein. The extent of biomaterial-mediated P1 and P2 exposure appears directly related to the severity of inflammatory responses to a test panel of biomaterials. Finally, thrombin-mediated conversion of fibrinogen to fibrin also exposes both P1 and P2 epitopes. These observations may help explain both the inflammation caused by many types of implanted biomaterials and that which occurs naturally following thrombotic events. (Blood. 2001;98:1231 1238) PMID- 11493476 TI - Characterization of immunologic tolerance induced by transfusion of UV-B- irradiated allogeneic mononuclear leukocytes. AB - Transfusions of UV-B--irradiated peripheral blood mononuclear cells (UV-B--PBMCs) from BALB/c (H-2(d)) mice into CBA (H-2(k)) mice can induce humoral immune tolerance to H-2(d) antigens, and the induced tolerance is partially mediated by negative regulatory PBMCs. To further identify which subset of spleen mononuclear leukocytes (MNLs) in the tolerant CBA mice is responsible for the negative regulatory activity, adoptive transfer experiments were conducted using spleen MNLs from the tolerant CBA mice. Results showed that only CD4(+) T cells could transfer the negative regulatory activity in a dose-dependent manner. This negative regulatory activity was significantly reduced when CD25(+) helper T cells were removed. Further study suggested that inhibition of IL-12 production by UV-B--irradiated PBMCs played a role in the induction of immune tolerance. In vitro study of the cytokine production profile by CBA CD4(+) T cells, after stimulation with gamma-irradiated BALB/c spleen cells, revealed an enhanced production of the type 2 T-cell cytokines after tolerance induction. Induction of tolerance also prevented the development of cytotoxic T cells in CBA mice against BALB/c MNLs. Adoptive transfer study suggested that the cellular immune tolerance was also mediated by CD4(+) negative regulatory T cells. The induced immune tolerance was nullified after 400 cGy sublethal gamma irradiation. These results suggest that the ex vivo study of cytokine production by T cells may be used to monitor tolerance induction and the selection of gamma radiation dose is critical for potential clinical application of the tolerance induced by UV-B--PBMCs. (Blood. 2001;98:1239-1245) PMID- 11493477 TI - Host marrow stem cell potential and engraftability at varying times after low dose whole-body irradiation. AB - High levels of chimerism in syngeneic BALB/c transplants were reported when hosts were exposed to 1 Gy (100 cGy) whole body irradiation (WBI) and infused with 40 x 10(6) marrow cells. The recovery of host stem cells and alterations of enhanced host engraftability at varying times after 1 Gy WBI have now been evaluated in this study. Male BALB/c marrow (40 x 10(6) cells) was infused into female BALB/c hosts immediately or at 6, 12, and 24 weeks after 1 Gy WBI of host female BALB/c mice; engraftment percentages 8 weeks after cell injection at week 0, 6, 12, or 24 were 68% +/- 12%, 45% +/- 15%, 51% +/- 12%, or 20% +/- 8%, respectively. Eight week engraftment levels in nonirradiated hosts average 7.7%. Conversely, engraftable stem cells measured at 8 weeks postengraftment in 1 Gy--exposed hosts were reduced to 8.6% +/- 3% of nonirradiated mice at time 0, 35% +/- 12% 6 weeks later, 49% +/- 10% at 3 months, and 21% +/- 7% at 6 months. Engraftment was still increased and stem cell decreased 1 year after 1 Gy. Furthermore, the primary cells transplanted into 1 Gy hosts can be serially transplanted, and the predominant effect of 1 Gy is directly on engrafting stem cells and not through accessory cells. These data show that transplantation in 1 Gy mice may be delayed until recovery of hematopoiesis, suggesting strategies in allogeneic transplantation to avoid the adverse effects of cytokine storm. The incomplete recovery of engraftable stem cells out to 12 months indicates that stem cell expansion, especially in patients previously treated with radiomimetic drugs, may not be feasible. (Blood. 2001;98:1246-1251) PMID- 11493478 TI - Antibodies from patients with heparin-induced thrombocytopenia stimulate monocytic cells to express tissue factor and secrete interleukin-8. AB - Thrombosis is a life-threatening complication that occurs in a subset of patients with heparin-induced thrombocytopenia (HITT). The pathogenic mechanisms underlying the variable occurrence of thrombosis in HITT is poorly understood. It was hypothesized that monocyte activation leading to tissue factor expression may play a role in promoting a thrombogenic state in HITT. This study demonstrates that a human platelet factor 4 (PF4)/heparin-specific murine monoclonal antibody (KKO) binds to peripheral blood-derived human monocytes in a PF4-dependent manner. KKO and antibodies from patients with HITT induce monocytes to synthesize and secrete interleukin-8 and induce cell-surface procoagulant activity, which is abrogated following treatment with antihuman tissue factor antibody. The findings suggest a novel mechanism by which PF4/heparin antibodies may promote a hypercoagulable state in patients with HITT. (Blood. 2001;98:1252-1254) PMID- 11493479 TI - Levels of vascular endothelial growth factor are elevated in patients with obstructive sleep apnea--hypopnea syndrome. AB - To better understand how humans adapt to hypoxia, the levels of hemoglobin (Hb), serum erythropoietin (Epo), and vascular endothelial growth factor (VEGF) were measured in 106 patients with severe obstructive sleep apnea-hypopnea syndrome. The results indicated that temporal hypoxic stimulation increases Hb. Furthermore, a minor increase in Epo and a substantial increase in VEGF were found. The induction in patients with severe sleep apnea was greater than that reported in other types of hypoxia. (Blood. 2001;98:1255-1257) PMID- 11493480 TI - Natural history of congenital dyserythropoietic anemia type II. AB - Congenital dyserythropoietic anemia type II (CDA-II) is an autosomal recessive disease characterized by anemia, jaundice, splenomegaly, and erythroblast multinuclearity. The natural history of the disease is unknown. The frequency, the relevance of complications, and the use of splenectomy are poorly defined. This study examined 98 patients from unrelated families enrolled in the International Registry of CDA-II. Retrospective data were obtained using an appropriate questionnaire. The mean age at presentation was 5.2 +/- 6.1 years. Anemia was present in 66% and jaundice in 53.4% of cases. The mean age at correct diagnosis was 15.9 +/- 11.8 years. Twenty-three percent of patients for whom data were available developed anemia during the neonatal period, and 10 of these individuals required transfusions. Splenectomy produced an increased hemoglobin (P <.001) and a reduced bilirubin level (P =.007) in comparison with values before splenectomy. Preliminary data indicate that iron overload occurs irrespective of the hemochromatosis genotype. (Blood. 2001;98:1258-1260) PMID- 11493481 TI - A novel delta beta fusion gene expresses hemoglobin A (HbA) not Hb Lepore: Senegalese delta(0)beta(+) thalassemia. AB - This study identified and characterized a novel delta beta fusion gene in which the delta-globin gene promoter is linked to intact beta-globin coding sequences in a Senegalese family. It results from a 7.4-kb deletion that removes the delta globin coding sequences, the delta beta intergenic region as well as the beta globin gene promoter and causes delta(0)beta(+) thalassemia with hemoglobin A expressed at the 11% to 15% range. The phenotype of this naturally occurring delta beta hybrid gene not only clarifies, in an in vivo context, the respective strength of delta- and beta-globin gene promoters, but also emphasizes the importance of beta-globin intragenic sequences in the expression of beta-globin chains. (Blood. 2001;98:1261-1263) PMID- 11493482 TI - A novel gene, NSD1, is fused to NUP98 in the t(5;11)(q35;p15.5) in de novo childhood acute myeloid leukemia. AB - The recurrent translocation t(5;11)(q35;p15.5) associated with a 5q deletion, del(5q), has been reported in childhood acute myeloid leukemia (AML). We report the cloning of the translocation breakpoints in de novo childhood AML harboring a cryptic t(5;11)(q35;p15.5). Fluorescence in situ hybridization (FISH) analysis demonstrated that the nucleoporin gene (NUP98) at 11p15.5 was disrupted by this translocation. By using 3'--rapid amplification of complementary DNA ends (3' RACE) polymerase chain reaction, we identified a chimeric messenger RNA that results in the in-frame fusion of NUP98 to a novel gene, NSD1. The NSD1 gene has 2596 amino acid residues and a 85% homology to the murine Nsd1 with the domain structure being conserved. The NSD1 gene was localized to 5q35 by FISH and is widely expressed. The reciprocal transcript, NSD1-NUP98, was also detected by reverse transcriptase--polymerase chain reaction. This is the first report in which the novel gene NSD1 has been implicated in human malignancy. (Blood. 2001;98:1264-1267) PMID- 11493483 TI - SH2D1A mutations in Japanese males with severe Epstein-Barr virus--associated illnesses. AB - X-linked lymphoproliferative disease (XLP), a genetic disorder characterized by immunodeficiency to Epstein-Barr virus (EBV) infection, has been linked to mutations in the SH2D1A gene. To search for the occurrence of SH2D1A mutations in Japan, we performed genetic analysis of the SH2D1A gene in 40 males presenting with severe EBV-associated illnesses, including fulminant infectious mononucleosis, EBV-positive lymphoma, and severe chronic active EBV infection. SH2D1A mutations were detected in 10 of these 40 patients. Five of these 10 cases were sporadic. Patients with SH2D1A mutations displayed severe acute infectious mononucleosis with hyperimmunoglobulin M, hypogammaglobulinemia, and B-cell malignant lymphoma. By contrast, chronic active EBV infection was not associated with SH2D1A mutations. XLP survivors exhibited normal levels of circulating EBV DNA during convalescence, suggesting that SH2D1A protein is not directly responsible for control of EBV replication. Thus, genetic analysis of the SH2D1A gene is particularly useful in the diagnosis of sporadic cases and carriers of XLP. (Blood. 2001;98:1268-1270) PMID- 11493485 TI - Sedation and analgesia-which way is best? PMID- 11493484 TI - Nitric oxide as mediator, marker and modulator of microvascular damage in ARDS. PMID- 11493486 TI - BJA concise - a step too far. PMID- 11493487 TI - Sedative and analgesic practice in the intensive care unit: the results of a European survey. AB - Sedation and analgesia are important aspects of patient care on the intensive care unit (ICU), yet relatively little information is available on common sedative and analgesic practice. We sought to assess international differences in the prescription of sedative and analgesic drugs in western European ICUs by means of a short, self-administered questionnaire. Six hundred and forty-seven intensive care physicians from 16 western European countries replied to the questionnaire. Midazolam was used as a sedative often or always by 63% of respondents and propofol by 35%. There were considerable international variations, with midazolam being preferred over propofol in France, Germany, the Netherlands, Norway and Austria. For analgesia, the drugs most commonly used were morphine (33%), fentanyl (33%) and sufentanil (24%). Morphine was preferred over fentanyl and sufentanil in Norway, UK and Ireland, Sweden, Switzerland, the Netherlands, and Spain and Portugal. Fentanyl was preferred in France, Germany and Italy. Sufentanil was preferred in Belgium and Luxemburg and in Austria. Multivariate analysis showed that the combination of midazolam with fentanyl was most often used in France; propofol with morphine in Sweden, the UK and Ireland, and Switzerland; midazolam with morphine in Norway; and propofol with sufentanil in Belgium and Luxemburg, Germany and Italy. The use of a sedation scale varied from 72% in the UK and Ireland to 18% in Austria. When used, the most common sedation scale was the Ramsay scale. This study demonstrates substantial international differences in sedative and analgesic practices in western European ICUs. PMID- 11493488 TI - Effects of desflurane on cerebral autoregulation. AB - The aim of this study was to determine the effects of desflurane, at 1 and 1.5 MAC, on cerebral autoregulation. Data were analysed from eight patients undergoing non-neurosurgical procedure. The blood flow velocity in the middle cerebral artery was measured by transcranial Doppler ultrasound and cerebral autoregulation was assessed by the transient hyperaemic response test. Partial pressure of the end-tidal carbon dioxide (PE'(CO(2))) and mean arterial pressure were measured throughout the study. Anaesthesia was induced with propofol and was maintained with desflurane at end-tidal concentrations of 7.4% (1 MAC) or 10.8% (1.5 MAC). The order of administration of the desflurane concentrations was determined randomly and a period of 15 min was allowed for equilibration at each concentration. The transient hyperaemic response tests were performed before induction of anaesthesia and after equilibration with each concentration of desflurane. An infusion of phenylephrine was used to maintain pre-induction mean arterial pressure and ventilation was adjusted to maintain the pre-induction value of PE'(CO(2)) throughout the study. Two indices derived from the transient hyperaemic response test (the transient hyperaemic response ratio and the strength of autoregulation) were used to assess cerebral autoregulation. Desflurane resulted in a marked and significant impairment in cerebral autoregulation; at concentrations of 1.5 MAC, autoregulation was almost abolished. PMID- 11493489 TI - Association of airway obstruction, sleep, and phasic abdominal muscle activity after upper abdominal surgery. AB - We recorded nasal gas flow, sleep stage, and abdominal muscle EMG pattern in 11 patients throughout the night after abdominal surgery, to examine the association between phasic activity of the abdominal muscles, sleep stage, and flow disturbance. We used a miniaturized data logging system, and obtained satisfactory records in eight patients. The data were divided into 30-s epochs. Each epoch was classified as either awake or asleep. The epochs were also classified for the presence of phasic activity in the external oblique abdominal muscle, and for evidence of airway obstruction. Association between these features was tested by a quasi likelihood log linear model. Values given are median (quartiles) for the eight subjects. Sleep occurred for 62 (46-69)% of the study time. During sleep, inspiratory flow was normal for 69 (48-81)% of the time, whereas during wakefulness, the flow pattern was normal for 51 (28-77)% of the time. Phasic activity was present 16 (12-25)% of the time during sleep and 24 (19-37)% of the time during wakefulness (P<0.001). In the awake state, when breathing was normal, phasic activity was present 16 (11-30)% of the time. When breathing was obstructed, phasic activity was present 38 (25-44)% of the time (P<0.001). These surprising findings suggest that sleep may be seriously disturbed by airway obstruction, so that a stable sleep state is not reached. We could not confirm previous findings that disturbed breathing in post-operative patients only occurs during sleep. PMID- 11493490 TI - Pulmonary scintigraphy for diagnosis of aspiration during intravenous propofol anaesthesia for colonoscopy. AB - A specific technique for detection of pulmonary aspiration during the perioperative period is lacking. In this study, we developed a scintigraphic method for its diagnosis. Technetium 99m sulphur colloid was given orally 2 h before an i.v. infusion of propofol in patients undergoing elective colonoscopy. During the procedure, patients were spontaneously breathing 100% oxygen via a face mask. After recovery from anaesthesia, patients had a chest scinti-scan. As a control group, 10 healthy men were studied. The lung scan was considered positive if any tracer activity greater than background level was detected in the lung field. Among 96 patients studied, three patients had a positive chest scinti scan. One of the three patients developed pneumonia while the other two remained asymptomatic. In none of the control asymptomatic group was tracer detected in the chest. We suggest that this technique is specific and can be used as a tool to assess the risk of pulmonary aspiration during different anaesthetic procedures. PMID- 11493491 TI - Manoeuvres used to clear the airway during fibreoptic intubation. AB - Fibreoptic orotracheal endoscopy under general anaesthesia may be more difficult to perform if the upper airway cannot be fully cleared. We have studied the effectiveness of jaw thrust, lingual traction and the application of both manoeuvres simultaneously, in opening up the orolaryngeal airspace in 30 ASA group 1 or 2 patients aged between 16 and 70 yr undergoing elective general surgery requiring orotracheal intubation. Airway clearance was assessed fibreoptically at soft palate level by observing whether or not the uvula or soft palate was apposed to the base of the tongue, and at epiglottic level by observing whether or not the epiglottis was apposed to the posterior pharyngeal wall. Lingual traction with Duval's forceps cleared the tongue away from the uvula and soft palate significantly more times than did jaw thrust (P<0.05). Jaw thrust cleared the epiglottis away from the posterior pharyngeal wall more frequently than did lingual traction (P=0.052). Applying both jaw thrust and lingual traction simultaneously cleared the airway at both soft palate and epiglottic level in every patient. When used alone, jaw thrust and lingual traction fail to produce full airway clearance in a significant number of patients. Combined jaw thrust and lingual traction clears the airway more effectively but requires two assistants. PMID- 11493492 TI - A comparison of cardiac output derived from the arterial pressure wave against thermodilution in cardiac surgery patients. AB - In three clinical centres, we compared a new method for measuring cardiac output with conventional thermodilution. The new method computes beat-to-beat cardiac output from radial artery pressure by simulating a three-element model of aortic input impedance, and includes non-linear aortic mechanical properties and a self adapting systemic vascular resistance. We compared cardiac output by continuous model simulation (MF) with thermodilution cardiac output (TD) in 54 patients (18 female, 36 male) undergoing coronary artery bypass surgery. We made three or four conventional thermodilution estimates spread equally over the ventilatory cycle. In 490 series of measurements, thermodilution cardiac output ranged from 2.1 to 9.3, mean 5.0 litre min(-1). MF differed +0.32 (1.0) litre min(-1) on average with limits of agreement of -1.68 and +2.32 litre min(-1). Differences decreased when the first series of measurements in a patient was used to calibrate the model. In 436 remaining series, the mean difference became -0.13 (0.47) litre min(-1) with limits of agreement of -1.05 and +0.79 litre min(-1). When consecutive measurements were made, the change was greater than 0.5 litre min( 1), on 204 occasions. The direction of change was the same with both methods in 199. The difference between the methods remained near zero during surgery suggesting that a single calibration per patient was adequate. Aortic model simulation with radial artery pressure as input reliably monitors changes in cardiac output in cardiac surgery patients. Before calibration, the model cannot replace thermodilution, but after calibration the model method can quantitatively replace further thermodilution estimates. PMID- 11493493 TI - Continuous intra-jugular venous blood-gas monitoring with the Paratrend 7 during hypothermic cardiopulmonary bypass. AB - We measured the accuracy of the continuous intra-vascular blood-gas monitoring system (Paratrend 7, PT7) placed in the jugular venous bulb in 18 adult patients having cardiac or aortic surgery with hypothermic cardiopulmonary bypass (CPB). After induction of anaesthesia, a PT7 sensor was inserted through a 20-gauge venous catheter into the right jugular venous bulb. Blood samples were drawn from the venous catheter and measured with a blood gas analyser (BGA). Five to eight paired measurements using the PT7 and blood samples were made per patient, and bias and precision were calculated for each patient using the Bland-Altman method. The ranges for the blood sample measurements were: pH 7.12 to 7.59, PCO(2) 3.7 to 9.6 kPa, PO(2) 3.5 to 16.0 kPa, oxygen saturation 40 to 99%, bicarbonate 18.6 to 34.4 mmol l(-1), and base excess -7.8 to 12.5 mmol l(-1). Bias and precision values were 0.014/0.071 for pH, 0/0.90 kPa for PCO(2), and 0.16/1.18 kPa for PO(2). These values were comparable with those previously made on arterial blood. However, precision for oxygen saturation in each patient varied 2.3 to 23.6% (95% CI: 6.3 to 12.9%), which was unsatisfactory for clinical measurements. Deep hypothermia ( approximately 19.6 degrees C) and marked haemodilution ( approximately 13.5%) during CPB did not influence the reliability of the PT7 sensor. Thus, we concluded that continuous intra-jugular venous blood gas monitoring is clinically feasible using the PT7 and may provide valuable information during CPB. PMID- 11493494 TI - Cerebral hypoperfusion in immediate postoperative period following coronary artery bypass grafting, heart valve, and abdominal aortic surgery. AB - Perioperative levels of jugular bulb oxyhaemoglobin saturation (Sj(O(2))) and lactate concentration (Lj), and postoperative duration of Sj(O(2))<50% were compared between patients undergoing coronary artery bypass grafting (CABG) (n=86), heart valve (n=14) and abdominal aortic (n=16) surgery. Radial artery and jugular bulb blood samples were aspirated after induction of anaesthesia, during re-warming on cardiopulmonary bypass (CPB) (36 degrees C), on arrival in the intensive care unit (ICU) and, subsequently, at 1, 2 and 6 h after ICU admission. Most patients having heart surgery were hypocapnic at 36 degrees C on CPB. Following CABG and heart valve surgery, many patients were hypocapnic whereas after abdominal aortic surgery, most were hypercapnic. During CPB and postoperatively, Sj(O(2)) and Lj were significantly correlated to Pa(CO(2)) and the arterial concentration of lactate (La) respectively (P<0.05). After correction for arterial carbon dioxide tension (Pa(CO(2))) and La, there were no significant changes in Sj(O(2)) or Lj on CPB. Postoperatively, having corrected for Pa(CO(2)), there were significant effects on Sj(O(2)) over all groups as a result of time from surgery (P<0.001) and its interaction with operation type (P<0.001). Following correction for La, there were no postoperative effects on Lj. No significant differences (P=0.2) in duration of Sj(O(2))<50% existed between patients undergoing CABG (1054 (82) min), abdominal aortic (893 (113) min) and heart valve (1073 (91) min) surgery. The lack of significant reciprocal effects on Lj combined with the frequency of hypocapnia and strong influence of Pa(CO(2))()on Sj(O(2)), suggest that Sj(O(2))<50% during CPB and after cardiac surgery represents hypoperfusion as a consequence of hypocapnia rather than cerebral ischaemia. PMID- 11493495 TI - Do physiological changes in pregnancy change defibrillation energy requirements? AB - Resuscitation during pregnancy is uncommon, with approximately 70 deaths occurring during pregnancy in the UK per annum. Physiological changes during pregnancy may affect transthoracic impedance (TTI), which determines transmyocardial current. Increased blood volume, cardiomegaly, haemodilution, changes in lung volume and changes in thoracic volume may alter impedance in ways that are difficult to predict. We measured TTI at term and after delivery once physiological changes had resolved. Mean (SD) TTI was 91.3 (15.8) Omega at term and 91.6 (11.8) Omega 6-8 weeks after delivery; the difference was not statistically significant. We conclude that current energy requirements for adult defibrillation are appropriate for use during pregnancy. PMID- 11493496 TI - Homozygous and heterozygous Arg614Cys mutations (1840C-->T) in the ryanodine receptor gene co-segregate with malignant hyperthermia susceptibility in a German family. AB - The determination of susceptibility to malignant hyperthermia (MH) by genetic investigation is a controversial issue because of the genetic heterogeneity of this disorder. The requirement for such an approach in MH diagnosis is a strong correlation between MH-associated genetic abnormalities and phenotypic findings in the in vitro contracture test (IVCT). After a severe clinical MH crisis during general anaesthesia a patient was diagnosed by the IVCT in which susceptibility to MH was confirmed. Genetic screening for MH-related mutations in the RYR1 gene revealed the presence of a homozygous 1840C-->T base exchange (Arg614Cys substitution) in this patient. A specific search for this defect in 20 relatives led to the identification of a total of 11 Arg614Cys mutations. Of these, 10 were heterozygous (including both parents) and one was homozygous (sister). Further IVCTs were subsequently performed on the parents of the index patient, the homozygous sister and all relatives who did not carry the Arg614Cys in order to determine the genotype/phenotype correlation. After analysing these data, and because of the strong correlation between clinical, phenotypic, and genetic results in the index patient, we assigned the diagnosis 'MHS' to all the remaining Arg614Cys mutation carriers of that family without performing the IVCT. PMID- 11493497 TI - Isolated reduction of haematocrit does not compromise in vitro blood coagulation. AB - Low haematocrit values are generally well tolerated in terms of oxygen transport but a low haematocrit might interfere with blood coagulation. We thus sampled 60 ml of blood in 30 healthy volunteers. The blood was centrifuged for 30 min at 2000 g and separated into plasma, which contained the platelet fraction, and packed red blood cells. The blood was subsequently reconstituted by combining the entire plasma fraction with a mixture of packed red blood cells, 0.9% saline, so that the final haematocrit was either 40, 30, 20, or 10%. Blood coagulation was assessed by computerized Thrombelastograph analysis. Data were compared using repeated measures analysis of variance and post-hoc paired t-tests with Bonferroni correction. Decreasing the haematocrit from 40 to 10% resulted in a shortening of reaction time (r) and coagulation time (k), and an increase in angle alpha, maximum amplitude (MA) and clot strength (G) (all P<0.02). This pattern represents acceleration of blood coagulation with low haematocrit values. The isolated reduction in haematocrit, therefore, does not compromise in vitro blood coagulation. PMID- 11493498 TI - A reduction in type and screen: preoperative prediction of RBC transfusions in surgery procedures with intermediate transfusion risks. AB - In many patients, a 'type and screen' procedure is routinely performed before surgery. However, most patients are not transfused after all. Can we predict, which surgical patients will and will not be transfused, to reduce the number of these investigations? We studied 1482 consecutive surgical patients with intermediate risk for transfusion. Multivariate logistic regression modelling and the area under the Receiver Operating Characteristic curve (ROC area) were used to quantify how well age, gender, surgical procedure, emergency or elective surgery and anaesthetic technique predicted transfusion, and whether the preoperative haemoglobin concentration had added predictive value. Gender, age > or =70 yr, and type of surgery were independent predictors of transfusion, with a ROC area of 0.75 (95% CI: 0.72-0.79). Validating this model with an easily used prediction rule in a second patient population yielded a ROC area of 0.70 (95% CI: 0.63-0.77). With this rule type and screen could correctly be withheld in 35% of these patients. In the remaining 65% of the patients, a further reduction in type and screen investigations of 15% could be achieved using the preoperative haemoglobin concentration. Using a simple prediction rule, preoperative type and screen investigations in patients who have to undergo surgery procedures with intermediate transfusion risk can be avoided in about 50%. This may reduce patient burden and hospital costs (on average: 3 million US$ per 100 000 procedures). PMID- 11493499 TI - Isoflurane-induced protection against myocardial stunning is independent of adenosine 1 (A(1)) receptor in isolated rat heart. AB - Volatile anaesthetics can pharmacologically enhance the recovery of stunned myocardium, but the mechanism is still unknown. This study sought to determine whether isoflurane attenuates myocardial stunning, and whether the myocardial protection of isoflurane is mediated by adenosine A(1) receptors. Five groups (n=8) of isolated rat hearts were studied in the Langendorff apparatus. The control groups underwent 20-min ischaemia with or without adenosine receptor antagonist (DPCPX, A(1)()selective) treatment (Cont group and DPCPX group). In the isoflurane groups, isoflurane (1.5 MAC) was present throughout the experiment (Iso group) and DPCPX (200 nM) was administered from 10 min before ischaemia (Iso+DPCPX(pre-I) group) or the beginning of reperfusion (Iso+DPCPX(post-I) group) to the end of experiment. The isoflurane groups had a lower end-diastolic pressure than the control groups (P<0.05). Developed pressure recovered to 77, 76, and 82% in Iso, Iso+DPCPX(pre-I) and Iso+DPCPX(post-I) groups, respectively (P<0.05 compared with control groups). LV+dp/dt(max) recovered to 53, 86, 81, 84, and 60% of pre-ischaemic values in Cont, Iso, Iso+DPCPX(pre-I), Iso+DPCPX(post I), and DPCPX groups. LV-dp/dt(min) recovered to 55, 84, 83, 81, and 62%, respectively. Both LV+dp/dt(max) and LV-dp/dt(min) were significantly different (P<0.05) between control and isoflurane groups during reperfusion. There were no significant differences among the isoflurane groups. Our data show that isoflurane enhances the post-ischaemic functional recovery of isolated rat heart and that block of A(1) receptors does not abolish the beneficial effects of isoflurane. We conclude that A(1)()receptors are not involved in isoflurane induced myocardial protection in the isolated rat heart. PMID- 11493501 TI - Microvessel damage in acute respiratory distress syndrome: the answer may not be NO. PMID- 11493500 TI - Aerosolized surfactant therapy for endotoxin-induced experimental acute respiratory distress syndrome in rats. AB - We have compared the effects of inhalation of aerosolized surfactant on experimental acute respiratory distress syndrome. Escherichia coli endotoxin (55 (SD 20) mg kg(-1)) was injected into the tracheas of 36 adult rats anaesthetized and mechanically ventilated with pure oxygen. When the Pa(O(2)) decreased to 11.3 (3.3) kPa, the animals were randomly subjected to inhalation of aerosolized modified natural surfactant (MNS) for 0 min (control group), 30, 60, and 120 min. In the control group, Pa(O(2)) remained below 12 kPa for 180 min. In the groups receiving inhalation of surfactant for 30 and 60 min, Pa(O(2)) increased but decreased soon after termination of the inhalation. In contrast, Pa(O(2)) of the group receiving inhalation of surfactant for 120 min continued to increase, reaching 52.1 (12.5) kPa at 180 min (P<0.05 vs control). Thus, we conclude that improvement in gas exchange as a result of inhalation of MNS depends on the duration of inhalation. PMID- 11493502 TI - Influence of sepsis on minimum alveolar concentration of desflurane in a porcine model. AB - The effect of sepsis on the minimum alveolar concentration of desflurane (MAC(DES)) in humans and other animals has not been reported previously. The aim of this study was to test the hypothesis that sepsis might alter MAC(DES) in a normotensive septic porcine model. Twenty-four young healthy pigs were premedicated with ketamine 10 mg kg(-1 )i.m and then anaesthesia was established with propofol 3 mg kg(-1) and the trachea was intubated. Baseline MAC(DES) in each pig was evaluated by pinching with a haemostat applied for 1 min to a rear dewclaw. MAC(DES) was determined by changing desflurane concentrations stepwise until purposeful movement appeared. Pigs were randomly assigned to two groups of 12 animals: the saline group received a 1 h i.v. infusion of saline solution while the sepsis group received a 1 h i.v. infusion of live Pseudomonas aeruginosa. Epinephrine and hydroxyethylstarch were used to maintain normotensive and normovolaemic haemodynamic status. In both groups, MAC(DES) was evaluated 5 h after infusion. Significant increases in heart rate, cardiac output, mean pulmonary artery pressure and pulmonary vascular resistance occurred in the sepsis group. MAC(DES) was 9.2% (95% confidence interval (CI) 6.8-10.6%) for the saline group and 6.7% (95% CI: 4.7-10.4) for the sepsis group (P<0.05). These data indicate that MAC(DES) is significantly decreased in this normotensive hyperkinetic septic porcine model. PMID- 11493503 TI - Loss of volition and pain response during induction of anaesthesia with propofol or sevoflurane. AB - We compared the time to reach two anaesthetic end-points during induction of anaesthesia with a potent inhalation agent (sevoflurane) and an i.v. agent (propofol). We used a method to ensure steady breathing during inhalation induction, and measured loss of tone in the outstretched arm and loss of response to a painful stimulus. Thirty-eight female patients (age 39 (9) yr, weight 65 (11) kg, and height 165 (8) cm) (mean (SD)) were randomly allocated to receive either propofol or sevoflurane. The predicted induction dose of propofol, estimated from age and weight for each patient, was given at a rate of 1% of the induction dose per second, to a possible maximum of 2.5 times the predicted induction dose. Sevoflurane was given with an inhaled concentration of 8%, which was anticipated to cause loss of arm tone within 90-120 s. After loss of consciousness, we applied a painful electrical stimulus to a finger at 15-s intervals and measured the time to loss of motor response. The median times and interquartile values for loss of arm tone were 105 (88-121) s for sevoflurane and 65 (58-80) s for propofol. This was equivalent to 0.65 of the ED(50) of propofol. The time to loss of response to pain was 226 (169-300) s for sevoflurane. The variances of these three measurements were not significantly different, indicating that these dose-response relationships were similar. In contrast, only 11 of the patients given propofol lost the response to pain after 2.5xED(50) had been given. These results support previous evidence of substantial differences between anaesthetic end-points, and show that this evidence can be obtained using a simple and rapid method. PMID- 11493504 TI - Influence of nitrous oxide on induction of anaesthesia with sevoflurane. AB - Nitrous oxide is often used during inhalation induction of anaesthesia with sevoflurane. Although the value of using nitrous oxide during inhalation induction with other volatile anaesthetics has been studied, the popularity of sevoflurane induction and the different characteristics of this agent make a study of the combination of nitrous oxide with this agent of interest. We compared induction times, oxygenation, and excitatory events during inhalation induction of anaesthesia using sevoflurane, with and without nitrous oxide. We studied 64 female patients, randomly allocated to receive inhalation induction of anaesthesia using sevoflurane with or without 50% nitrous oxide in the fresh gas, using a co-axial breathing system (Mapleson D) and a fresh gas flow rate of 3-6 litre min(-1). Mean time to induction of anaesthesia (fall of an outstretched arm) was 102 s in both groups, but excitation (limb or head movement) was more frequent in those receiving nitrous oxide (10 patients) than in those receiving oxygen only (five patients) (P<0.05). Oxygenation was similar in both groups. We conclude that nitrous oxide confers no advantage when anaesthesia is induced with sevoflurane in this way. PMID- 11493505 TI - Air flow resistance of three heat and moisture exchanging filter designs under wet conditions: implications for patient safety. AB - Heat and moisture exchanging filters (HMEFs) can be blocked by secretions. We have studied HMEF performance under wet conditions to see which particular design features predispose to this complication. Dar Hygrobac-S (composite felt filter and cellulose exchanger), Dar Hygroster (composite pleated ceramic membrane and cellulose exchanger) and Pall BB22-15 (pleated ceramic membrane) HMEFs were tested. Saline retention, saline concealment, and changes in air flow resistance when wet were assessed. The cellulose exchanger in the composite Hygrobac-S and Hygroster retained saline, producing a 'tampon' effect, associated with bi directional air flow resistances in excess of the international standard of a 5 cm H(2)O pressure drop at 60 litre min(-1) air flow. Furthermore, high air flow resistances occurred before free saline was apparent within the transparent filter housing. The pleat only BB22-15 showed a significant increase in expiratory air flow resistance, but only after the presence of saline was apparent. These data imply that composite HMEFs with cellulose exchangers are more likely to block or cause excessive work of breathing as a result of occult accumulation of patient secretions than pleat only HMEFs. PMID- 11493506 TI - Touch contamination levels during anaesthetic procedures and their relationship to hand hygiene procedures: a clinical audit. AB - After different methods of hand preparation, volunteers rolled segments of sterile central venous catheter between their fingertips, and bacterial transfer was evaluated by standardized quantitative culture. The number of bacteria transferred differed between methods (P<0.001). Comparisons were made with the control group (no preparation at all; median, third quartile and maximum count=6.5, 24, 55). Bacterial transfer was greatly increased with wet hands (1227, 1932, 3254; P<0.001). It was reduced with a new rapid method, based on thorough drying with a combination of 10 s using a cloth towel followed by either 10 or 20 s with a hot-air towel (0, 3, 7 and 0, 4, 30, respectively; P=0.007 and 0.004, respectively). When asked to follow their personal routines, 10 consultant anaesthetists used a range of methods. Collectively, these were not significantly better than control (7.5, 15, 55; P=0.73), and neither was an air towel alone (2.5, 15, 80; P=0.176) nor the hospital's standard procedure (0, 1, 500; P=0.035). If hand preparation is needed, an adequate and validated method should be used, together with thorough hand drying. PMID- 11493507 TI - Primary pulmonary hypertension in pregnancy; a role for novel vasodilators. AB - We describe the case of a 28-week pregnant woman presenting with severe primary pulmonary hypertension (PPH). She had an elective Caesarean section under general anaesthesia at 32 weeks gestation. Pulmonary artery pressures (PAP) measured from a pulmonary artery catheter before anaesthesia were in excess of 100 mm Hg. Intraoperative nitric oxide was used to reduce PAP. After the delivery of a healthy infant PAP was controlled with nebulized iloprost and a prostacyclin infusion. Seven days later she was discharged from intensive care taking an oral calcium antagonist and warfarin. She developed intractable right heart failure and died 14 days after delivery. Despite increasing experience in the use of drugs to reduce PAP, the clinical course of pregnancy complicated by severe PPH is usually fatal. PMID- 11493508 TI - Central venous catheterization and fatal cardiac tamponade. AB - Cardiac tamponade is a poorly recognized complication of central venous catheterization associated with a high mortality. We present a case of fatal cardiac tamponade after intra- pericardial infusion of total parenteral nutrition in a patient who had two central venous catheters. We suggest that catheter tip position should always be confirmed before use of a catheter. Tamponade should be suspected in a patient who deteriorates when a central venous catheter is used and resuscitation via the catheter should be avoided. PMID- 11493509 TI - Ruptured superior thyroid artery from central vein cannulation: treatment by coil embolization. AB - Central vein cannulation is associated with several complications, some of which may be fatal. This case report describes a life-threatening complication after insertion of a central venous line. An obese female patient with end-stage renal failure due to nephrotic syndrome developed a huge neck swelling and sudden airway obstruction after attempted cannulation of the internal jugular vein for haemodialysis. Tracheal intubation was achieved using a gum-elastic bougie. Investigations revealed abnormal blood clotting. The coagulopathy was treated, but the neck swelling continued to increase in size. Carotid angiography showed a ruptured right superior thyroid artery. The haemorrhage was controlled by coil embolization of the artery. This case report demonstrates the usefulness of the gum-elastic bougie in the presence of a difficult airway and of interventional radiology in the management of vascular accidents. PMID- 11493510 TI - Two episodes of life-threatening anaphylaxis in the same patient to a chlorhexidine-sulphadiazine-coated central venous catheter. AB - Chlorhexidine allergy has been described in the literature, mainly in Japanese individuals. Most reactions have been limited to the skin, mild in severity and a result of chlorhexidine containing solutions such as 'Savlon' (Novartis Consumer Health, Horesham, UK). We describe what we believe is the first reported case of anaphylaxis in a European patient to a chlorhexidine- sulphadiazine-coated central venous catheter. PMID- 11493511 TI - Delayed diagnosis of cardiac tamponade following isolated blunt abdominal trauma. AB - Traumatic haemopericardium is an uncommon but life threatening condition. It is usually caused by penetrating cardiac injuries or cardiac rupture from blunt chest trauma. We report haemopericardium and cardiac tamponade in a young girl after blunt abdominal trauma. She presented with mild upper abdominal pain, tachycardia and hypotension having been kicked in the abdomen by a horse. No damage was found at laparotomy and she remained haemodynamically unstable. Further investigation found cardiac tamponade and haemopericardium. This was managed by insertion of a pericardial drain using transthoracic echocardiogram guidance, with later drainage in the operating theatre using guidance with a transoesophageal echocardiogram. PMID- 11493512 TI - Ipsilateral thoraco-lumbar anaesthesia and paravertebral spread after low thoracic paravertebral injection. AB - We report ipsilateral thoraco-lumbar anaesthesia and paravertebral spread of contrast after injection through a thoracic paravertebral catheter that was placed at the right T8-9 spinal level for pain management in a patient with multiple fractured ribs. We review the literature and describe the subendothoracic fascial communication between the thoracic paravertebral space and the retroperitoneal lumbar paravertebral region, which we propose, is the anatomical basis for ipsilateral thoraco-lumbar anaesthesia and paravertebral spread of contrast in our patient. PMID- 11493513 TI - Another feature of TURP syndrome: hyperglycaemia and lactic acidosis caused by massive absorption of sorbitol. AB - Endoscopic transurethral resection of the prostate (TURP) can be complicated by absorption of a large volume of irrigation fluid. The clinical features of this complication are referred as the TURP syndrome. We report a case where hyperglycaemia and lactic acidosis complicated the TURP syndrome caused by the massive absorption (approximately 15 litres) of a sorbitol- mannitol irrigation solution. The proposed mechanism is a type B lactic acidosis related to the metabolism of sorbitol. PMID- 11493515 TI - Abstracts of the Intensive Care Society and Riverside Group "State of the Art" meeting. London, United Kingdom, December 7-8, 2000. PMID- 11493516 TI - The Handlebars gene is required with Phantastica for dorsoventral asymmetry of organs and for stem cell activity in Antirrhinum. AB - In angiosperms, individual lateral organs and whole flowers may develop asymmetrically along their dorsoventral axes. Dorsoventral asymmetry of Antirrhinum leaves requires activity of the Phantastica gene and other factors acting redundantly with it. We describe the effects of a mutation in the Handlebars gene, identified as an enhancer of the phantastica mutant phenotype. Genetic analysis suggests that Handlebars functions redundantly with Phantastica to promote dorsal fate in lateral organs and to maintain activity of stem cells within shoot apical meristems. Handlebars appears dispensable in vegetative development but is needed for asymmetry of petals along the dorsoventral axis of the flower as a whole. This suggests that common mechanisms may control dorsoventral asymmetry in lateral organ primordia and in floral meristems. PMID- 11493517 TI - TGF-beta modulates programmed cell death in the retina of the developing chick embryo. AB - Programmed cell death (PCD) is a key phenomenon in the regulation of cell number in multicellular organisms. We have shown that reduction of endogenous transforming growth factor beta (TGF-beta) prevents apoptotic PCD of neurons in the developing peripheral and central nervous system, suggesting that TGF-beta is an important mediator of ontogenetic neuron death. Previous studies suggested that there are other pro-apoptotic molecules, nerve growth factor (NGF) and brain derived neurotrophic factor, that induce cell death in the nervous system. In the developing chick retina, NGF induces PCD by activation of the p75 receptor. We have studied the role of TGF-beta and its putative interdependence with NGF mediated PCD in the chick retina. We found that TGF-beta is present in the developing chick retina during the period of PCD and is essentially required to regulate PCD of retinal cells. TGF-beta 2, TGF-beta 3 and the ligand-binding TGF beta receptor can be detected immunocytochemically in the central retina, a region where apoptosis is most prominent during the early period of PCD. Application of a TGF-beta-neutralizing antibody to chick embryos in ovo resulted in a decrease in the number of TUNEL-positive cells and a reduction of free nucleosome levels. In terms of magnitude, reduction of PCD caused by the neutralization of endogenous TGF-beta was equivalent to that seen after anti-NGF application. Neutralization of both factors did not result in a further decrease in apoptosis, indicating that NGF and TGF-beta may act on the same cell population. Furthermore, neutralization of TGF-beta did not affect the expression of NGF or the p75-receptor. Our results suggest that TGF-beta and NGF are both required to regulate cell death in the chick retina in vivo. PMID- 11493518 TI - Requirements for the kit receptor tyrosine kinase during regeneration of zebrafish fin melanocytes. AB - Embryonic neural crest-derived melanocytes and their precursors express the kit receptor tyrosine kinase and require its function for their migration and survival. However, mutations in kit also cause deficits in melanocytes that make up adult pigment patterns, including melanocytes that re-establish the zebrafish fin stripes during regeneration. As adult melanocytes in mice and zebrafish are generated and maintained by stem cell populations that are presumably established during embryonic development, it has been proposed that adult phenotypes in kit mutants result from embryonic requirements for kit. We have used a temperature sensitive zebrafish kit mutation to show that kit is required during adult fin regeneration to promote melanocyte differentiation, rather than during embryonic stages to establish their stem cell precursors. We also demonstrate a transient role for kit in promoting the survival of newly differentiated regeneration melanocytes. PMID- 11493519 TI - A regulatory cascade of three homeobox genes, ceh-10, ttx-3 and ceh-23, controls cell fate specification of a defined interneuron class in C. elegans. AB - The development of the nervous system requires the coordinated activity of a variety of regulatory factors that define the individual properties of specific neuronal subtypes. We report a regulatory cascade composed of three homeodomain proteins that act to define the properties of a specific interneuron class in the nematode C. elegans. We describe a set of differentiation markers characteristic for the AIY interneuron class and show that the ceh-10 paired-type and ttx-3 LIM type homeobox genes function to regulate all known subtype-specific features of the AIY interneurons. In contrast, the acquisition of several pan-neuronal features is unaffected in ceh-10 and ttx-3 mutants, suggesting that the activity of these homeobox genes separates pan-neuronal from subtype-specific differentiation programs. The LIM homeobox gene ttx-3 appears to play a central role in regulation of AIY differentiation. Not only are all AIY subtype characteristics lost in ttx-3 mutants, but ectopic misexpression of ttx-3 is also sufficient to induce AIY-like features in a restricted set of neurons. One of the targets of ceh-10 and ttx-3 is a novel type of homeobox gene, ceh-23. We show that ceh-23 is not required for the initial adoption of AIY differentiation characteristics, but instead is required to maintain the expression of one defined AIY differentiation feature. Finally, we demonstrate that the regulatory relationship between ceh-10, ttx-3 and ceh-23 is only partially conserved in other neurons in the nervous system. Our findings illustrate the complexity of transcriptional regulation in the nervous system and provide an example for the intricate interdependence of transcription factor action. PMID- 11493520 TI - SDF-1 alpha induces chemotaxis and enhances Sonic hedgehog-induced proliferation of cerebellar granule cells. AB - The chemokine SDF-1 alpha (CXC12) and its receptor CXCR4 have been shown to play a role in the development of normal cerebellar cytoarchitecture. We report here that SDF-1 alpha both induces chemotactic responses in granule precursor cells and enhances granule cell proliferative responses to Sonic hedgehog. Chemotactic and proliferative responses to SDF-1 alpha are greater in granule cells obtained from cerebella of animals in the first postnatal week, coinciding with the observed in vivo peak in cerebellar CXCR4 expression. SDF-1 alpha activation of neuronal CXCR4 differs from activation of CXCR4 in leukocytes in that SDF-1 alpha induced calcium flux is activity dependent, requiring predepolarization with KCl or pretreatment with glutamate. However, as is the case in leukocytes, neuronal responses to SDF-1 alpha are all abolished by pretreatment of granule cells with pertussis toxin, suggesting they occur through G(alpha i) activation. In conclusion, SDF-1 alpha plays a role in two important processes of granule cell maturation - proliferation and migration - assisting in the achievement of appropriate cell number and position in the cerebellar cortex. PMID- 11493521 TI - Cortical upper layer neurons derive from the subventricular zone as indicated by Svet1 gene expression. AB - The cerebral cortex is composed of a large variety of different neuron types. All cortical neurons, except some interneurons, are born in two proliferative zones, the cortical ventricular (VZ) and subventricular (SVZ) zones. The relative contribution of both proliferative zones to the generation of the diversity of the cortical neurons is not well understood. To further dissect the underlying mechanism, molecular markers specific for the SVZ are required. Towards this end we performed a subtraction of cDNA libraries, generated from E15.5 and E18.5 mouse cerebral cortex. A novel cDNA, Svet1, was cloned which was specifically expressed in the proliferating cells of the SVZ but not the VZ. The VZ is marked by the expression of the Otx1 gene. Later in development, Svet1 and Otx1 were expressed in subsets of cells of upper (II-IV) and deep (V-VI) layers, respectively. In the reeler cortex, where the layers are inverted, Svet1 and Otx1 label precursors of the upper and deeper layers, respectively, in their new location. Interestingly, in the Pax6/small eye mutant, Svet1 activity was abolished in the SVZ and in the upper part of the cortical plate while the Otx1 expression domain remained unchanged. Therefore, using Svet1 and Otx1 as cell type-specific molecular markers for the upper and deep cortical layers we conclude that the Sey mutation affects predominantly the differentiation of the SVZ cells that fail to migrate into the cortical plate. The abnormality of the SVZ coincides with the absence of upper layer cells in the cortex. Taken together our data suggest that while the specification of deep cortical layers occurs in the ventricular zone, the SVZ is important for the proper specification of upper layers. PMID- 11493522 TI - Pax3 acts cell autonomously in the neural tube and somites by controlling cell surface properties. AB - Pax3 is a member of the paired-box-containing transcription factors. It is expressed in the developing somites, dorsal spinal cord, mesencephalon and neural crest derivatives. Several loss-of-function mutations are correlated with the Splotch phenotype in mice and Waardenburg syndrome in humans. Malformations include a lack of muscle in the limb, a failure of neural tube closure and dysgenesis of numerous neural crest derivatives. In this study we have used embryonic stem (ES) cells to generate a lacZ knock-in into the Pax3 locus. The Pax3 knock-in Splotch allele (Sp(2G)) was used to generate Pax3-deficient ES cells in order to investigate whether, in chimeric embryos, Pax3 is acting cell autonomously in the somites and the neural tube. We found that while Pax3 function is essential for the neuroepithelium and somites, a wild-type environment rescues mutant neural crest cells. In the two affected embryonic tissues, mutant and wild-type cells undergo segregation and do not intermingle. The contribution of mutant cells to the neural tube and the somites displayed temporal differences. All chimeric embryos showed a remarkable contribution of blue cells to the neural tube at all stages analyzed, indicating that the Pax3 deficient cells are not excluded from the neural epithelium while development proceeds. In contrast, this is not true for the paraxial mesoderm. The somite contribution of Pax3(-/-) ES cells becomes less frequent in older embryos as compared to controls with Pax3(+/-) ES cells. We propose that although Pax3 function is related to cell surface properties, its role may differ in various tissues. In fact, apoptosis was found in Pax3-deficient cells of the lateral dermomyotome but not in the neural tube. PMID- 11493523 TI - Binary specification of nerve cord and notochord cell fates in ascidian embryos. AB - In the ascidian embryo, the nerve cord and notochord of the tail of tadpole larvae originate from the precursor blastomeres for both tissues in the 32-cell stage embryo. Each fate is separated into two daughter blastomeres at the next cleavage. We have examined mechanisms that are responsible for nerve cord and notochord specification through experiments involving blastomere isolation, cell dissociation, and treatment with basic fibroblast growth factor (bFGF) and inhibitors for the mitogen-activated protein kinase (MAPK) cascade. It has been shown that inductive cell interaction at the 32-cell stage is required for notochord formation. Our results show that the nerve cord fate is determined autonomously without any cell interaction. Presumptive notochord blastomeres also assume a nerve cord fate when they are isolated before induction is completed. By contrast, not only presumptive notochord blastomeres but also presumptive nerve cord blastomeres forsake their default nerve cord fate and choose the notochord fate when they are treated with bFGF. When the FGF-Ras-MAPK signaling cascade is inhibited, both blastomeres choose the default nerve cord pathway, supporting the results of blastomere isolation. Thus, binary choice of alternative fates and asymmetric division are involved in this nerve cord/notochord fate determination system, mediated by FGF signaling. PMID- 11493524 TI - Otx genes are required for tissue specification in the developing eye. AB - Patterning of the vertebrate eye appears to be controlled by the mutual regulation and the progressive restriction of the expression domains of a number of genes initially co-expressed within the eye anlage. Previous data suggest that both Otx1 and Otx2 might contribute to the establishment of the different eye territories. Here, we have analysed the ocular phenotype of mice carrying different functional copies of Otx1 and Otx2 and we show that these genes are required in a dose-dependent manner for the normal development of the eye. Thus, all Otx1(-/-); Otx2(+/-) and 30% of Otx1(+/-); Otx2(+/-) genotypes presented consistent and profound ocular malformation, including lens, pigment epithelium, neural retina and optic stalk defects. During embryonic development, optic vesicle infolding was severely altered and the expression of pigment epithelium specific genes, such as Mitf or tyrosinase, was lost. Lack of pigment epithelium specification was associated with an expansion of the prospective neural retina and optic stalk territories, as determined by the expression of Pax6, Six3 and Pax2. Later in development the presumptive pigment epithelium region acquired features of mature neural retina, including the generation of Islet1-positive neurones. Furthermore, in Otx1(-/-); Otx2(+/-) mice neural retina cell proliferation, cell differentiation and apoptotic cell death were also severely affected. Based on these findings we propose a model in which Otx gene products are required for the determination and differentiation of the pigment epithelium, co-operating with other eye patterning genes in the determination of the specialised tissues that will constitute the mature vertebrate eye. PMID- 11493525 TI - Roles of retinoic acid receptors in early embryonic morphogenesis and hindbrain patterning. AB - Mutants mice carrying targeted inactivations of both retinoic acid receptor (RAR) alpha and RAR gamma (A alpha/A gamma mutants) were analyzed at different embryonic stages, in order to establish the timing of appearance of defects that we previously observed during the fetal period. We show that embryonic day (E)9.5 A alpha/A gamma embryos display severe malformations, similar to those already described in retinaldehyde dehydrogenase 2 null mutants. These malformations reflect early roles of retinoic acid signaling in axial rotation, segmentation and closure of the hindbrain; formation of otocysts, pharyngeal arches and forelimb buds; and in the closure of the primitive gut. The hindbrain of E8.5 A alpha/A gamma embryos shows a posterior expansion of rhombomere 3 and 4 (R3 and R4) markers, but fails to express kreisler, a normal marker of R5 and R6. This abnormal hindbrain phenotype is strikingly different from that of embryos lacking RAR alpha and RAR beta (A alpha/A beta mutants), in which we have previously shown that the territory corresponding to R5 and R6 is markedly enlarged. Administration of a pan-RAR antagonist at E8.0 to wild-type embryos cultured in vitro results in an A alpha/A beta-like hindbrain phenotype, whereas an earlier treatment at E7.0 yields an A alpha/A gamma-like phenotype. Altogether, our data suggest that RAR alpha and/or RAR gamma transduce the RA signal that is required first to specify the prospective R5/R6 territory, whereas RAR beta is subsequently involved in setting up the caudal boundary of this territory. PMID- 11493526 TI - Regulation of left-right asymmetry by thresholds of Pitx2c activity. AB - Although much progress has been made in understanding the molecular mechanisms regulating left-right asymmetry, the final events of asymmetric organ morphogenesis remain poorly understood. The phenotypes of human heterotaxia syndromes, in which organ morphogenesis is uncoupled, have suggested that the early and late events of left-right asymmetry are separable. The Pitx2 homeobox gene plays an important role in the final stages of asymmetry. We have used two new Pitx2 alleles that encode progressively higher levels of Pitx2c in the absence of Pitx2a and Pitx2b, to show that different organs have distinct requirements for Pitx2c dosage. The cardiac atria required low Pitx2c levels, while the duodenum and lungs used higher Pitx2c doses for normal development. As Pitx2c levels were elevated, the duodenum progressed from arrested rotation to randomization, reversal and finally normal morphogenesis. In addition, abnormal duodenal morphogenesis was correlated with bilateral expression of Pitx2c. These data reveal an organ-intrinsic mechanism, dependent upon dosage of Pitx2c, that governs asymmetric organ morphogenesis. They also provide insight into the molecular events that lead to the discordant organ morphogenesis of heterotaxia. PMID- 11493527 TI - Differential contributions of AF-1 and AF-2 activities to the developmental functions of RXR alpha. AB - We have engineered a mouse mutation that specifically deletes most of the RXR alpha N-terminal A/B region, which includes the activation function AF-1 and several phosphorylation sites. The homozygous mutants (RXR alpha af1(o)), as well as compound mutants that further lack RXR beta and RXR gamma, are viable and display a subset of the abnormalities previously described in RXR alpha-null mutants. In contrast, RXR alpha af1(o)/RAR(-/-)(alpha, beta or gamma) compound mutants die in utero and exhibit a large array of malformations that nearly recapitulate the full spectrum of the defects that characterize the fetal vitamin A-deficiency (VAD) syndrome. Altogether, these observations indicate that the RXR alpha AF-1 region A/B is functionally important, although less so than the ligand dependent activation function AF-2, for efficiently transducing the retinoid signal through RAR/RXR alpha heterodimers during embryonic development. Moreover, it has a unique role in retinoic acid-dependent involution of the interdigital mesenchyme. During early placentogenesis, both the AF-1 and AF-2 activities of RXR alpha, beta and gamma appear to be dispensable, suggesting that RXRs act as silent heterodimeric partners in this process. However, AF-2 of RXR alpha, but not AF-1, is required for differentiation of labyrinthine trophoblast cells, a late step in the formation of the placental barrier. PMID- 11493528 TI - Difference in XTcf-3 dependency accounts for change in response to beta-catenin mediated Wnt signalling in Xenopus blastula. AB - Wnt signalling functions in many tissues and during different stages of animal development to produce very specific responses. In early Xenopus embryos there is a dramatic change in response to Wnt signalling within only a few hours of development. Wnt signalling in very early embryos leads to a dorsalising response, which establishes the endogenous dorsal axis. Only a few hours later in development, almost the opposite happens: Xwnt-8 functions to pattern the embryonic mesoderm by promoting ventral and lateral mesoderm. The specificity of the response could conceivably be carried out by differential use of different signal transduction pathways, many of which have recently been described. We have found, however, that this dramatic shift in response to Wnt signalling in early Xenopus is not brought about by differential use of distinct signal transduction pathways. In fact beta-catenin, a downstream component of the canonical Wnt signal transduction pathway, functions not only in the early dorsalising response but also in the later ventrolateral-promoting response. Interaction of beta catenin with the XTcf-3 transcription factor is required for the early dorsalising activity. In contrast, our experiments suggest that late Wnt signalling in the ventrolateral mesoderm does not require a similar dependency of beta-catenin function on XTcf-3. Our results highlight the potential versatility of the canonical Wnt pathway to interact with tissue-specific factors downstream of beta-catenin, in order to achieve tissue-specific effects. PMID- 11493529 TI - Role of FGFs in the control of programmed cell death during limb development. AB - We have investigated the role of FGFs in the control of programmed cell death during limb development by analyzing the effects of increasing and blocking FGF signaling in the avian limb bud. BMPs are currently considered as the signals responsible for cell death. Here we show that FGF signaling is also necessary for apoptosis and that the establishment of the areas of cell death is regulated by the convergence of FGF- and BMP-mediated signaling pathways. As previously demonstrated, cell death is inhibited for short intervals (12 hours) after administration of FGFs. However, this initial inhibition is followed (24 hours) by a dramatic increase in cell death, which can be abolished by treatments with a BMP antagonist (Noggin or Gremlin). Conversely, blockage of FGF signaling by applying a specific FGF-inhibitor (SU5402) into the interdigital regions inhibits both physiological cell death and that mediated by exogenous BMPs. Furthermore, FGF receptors 1, 2 and 3 are expressed in the autopodial mesoderm during the regression of the interdigital tissue, and the expression of FGFR3 in the interdigital regions is regulated by FGFs and BMPs in the same fashion as apopotosis. Together our findings indicate that, in the absence of FGF signaling BMPs are not sufficient to trigger apoptosis in the developing limb. Although we provide evidence for a positive influence of FGFs on BMP gene expression, the physiological implication of FGFs in apoptosis appears to result from their requirement for the expression of genes of the apoptotic cascade. We have identified MSX2 and Snail as candidate genes associated with apoptosis the expression of which requires the combined action of FGFs and BMPs. PMID- 11493530 TI - Bves: prototype of a new class of cell adhesion molecules expressed during coronary artery development. AB - Bves is a protein expressed in cells of the developing coronary vascular system, specifically in the proepicardium, migrating epithelial epicardium, delaminated vasculogenic mesenchyme and vascular smooth muscle cells. Here, we show that Bves protein undergoes a dynamic subcellular redistribution during coronary vessel development. Bves is a membrane protein with three predicted transmembrane helices, an extracellular C terminus and an intracellular N terminus, and is confined to the lateral membrane compartment of epithelial cells. When epicardial cells are dissociated into single cells in vitro, Bves accumulates in a perinuclear region until cells make contact, at which time Bves is trafficked to the cell membrane. Bves accumulates at points of cell/cell contact, such as filopodia and cell borders, before the appearance of E-cadherin, suggesting an early role in cell adhesion. While Bves shares no homology with any known adhesion molecule, transfection of Bves into L-cells readily confers adhesive behavior to these cells. Finally, Bves antibodies inhibit epithelial migration of vasculogenic cells from the proepicardium. This study provides direct evidence that Bves is a novel cell adhesion molecule and suggests a role for Bves in coronary vasculogenesis. PMID- 11493531 TI - Lung hypoplasia and neonatal death in Fgf9-null mice identify this gene as an essential regulator of lung mesenchyme. AB - Mammalian lung develops as an evagination of ventral gut endoderm into the underlying mesenchyme. Iterative epithelial branching, regulated by the surrounding mesenchyme, generates an elaborate network of airways from the initial lung bud. Fibroblast growth factors (FGFs) often mediate epithelial mesenchymal interactions and mesenchymal Fgf10 is essential for epithelial branching in the developing lung. However, no FGF has been shown to regulate lung mesenchyme. In embryonic lung, Fgf9 is detected in airway epithelium and visceral pleura at E10.5, but is restricted to the pleura by E12.5. We report that mice homozygous for a targeted disruption of Fgf9 exhibit lung hypoplasia and early postnatal death. Fgf9(-/-) lungs exhibit reduced mesenchyme and decreased branching of airways, but show significant distal airspace formation and pneumocyte differentiation. Our results suggest that Fgf9 affects lung size by stimulating mesenchymal proliferation. The reduction in the amount of mesenchyme in Fgf9(-/-) lungs limits expression of mesenchymal Fgf10. We suggest a model whereby FGF9 signaling from the epithelium and reciprocal FGF10 signaling from the mesenchyme coordinately regulate epithelial airway branching and organ size during lung embryogenesis. PMID- 11493532 TI - Armadillo nuclear import is regulated by cytoplasmic anchor Axin and nuclear anchor dTCF/Pan. AB - Drosophila melanogaster Armadillo plays two distinct roles during development. It is a component of adherens junctions, and functions as a transcriptional activator in response to Wingless signaling. In the current model, Wingless signal causes stabilization of cytoplasmic Armadillo allowing it to enter the nucleus where it can activate transcription. However, the mechanism of nuclear import and export remains to be elucidated. In this study, we show that two gain of-function alleles of Armadillo activate Wingless signaling by different mechanisms. The S10 allele was previously found to localize to the nucleus, where it activates transcription. In contrast, the Delta Arm allele localizes to the plasma membrane, and forces endogenous Arm into the nucleus. Therefore, Delta Arm is dependent on the presence of a functional endogenous allele of arm to activate transcription. We show that Delta Arm may function by titrating Axin protein to the membrane, suggesting that it acts as a cytoplasmic anchor keeping Arm out of the nucleus. In axin mutants, Arm is localized to the nuclei. We find that nuclear retention is dependent on dTCF/Pangolin. This suggests that cellular distribution of Arm is controlled by an anchoring system, where various nuclear and cytoplasmic binding partners determine its localization. PMID- 11493533 TI - STAT1 mediates the increased apoptosis and reduced chondrocyte proliferation in mice overexpressing FGF2. AB - Unregulated FGF receptor signaling results in bone malformations that affect both endochondral and intramembranous ossification, and is the basis for several genetic forms of human dwarfism. FGF signaling inhibits chondrocyte proliferation and we have previously shown that the transcription factor STAT1 mediates the growth inhibitory effect of FGF in vitro. We provide genetic evidence that STAT1 is a modulator of the negative regulation of bone growth by FGF in vivo. We crossed Stat1(-/-) mice with a transgenic mouse line overexpressing human FGF2 (TgFGF). TgFGF mice exhibit phenotypes characterized by chondrodysplasia and macrocephaly, which affect endochondral and intramembranous ossification. We found that the chondrodysplasic phenotype of these mice results both from reduced proliferation and increased apoptosis of growth plate chondrocytes. Loss of STAT1 function in TgFGF mice led to a significant correction of the chondrodysplasic phenotype, but did not affect the skull malformations. The reduced proliferation of TgFGF growth plate chondrocytes, as well as their excessive apoptosis, were restored to near-normal levels in the absence of STAT1 function. Unregulated FGF signaling in TgFGF mice also induced apoptosis in calvarial osteoblasts that was not, however, corrected by the absence of STAT1. Detailed analysis of Stat1(-/-) growth plates uncovered a transient phenotype, characterized by an expansion of the proliferative zone and by acceleration of longitudinal bone growth, that attenuated as the animals grew older. These results document an essential role for STAT1 in FGF-mediated regulation of cell growth that is specific to the epiphyseal growth plate. PMID- 11493534 TI - The zebrafish space cadet gene controls axonal pathfinding of neurons that modulate fast turning movements. AB - All vertebrates depend on neural circuits to produce propulsive movements; however, the contribution of individual neural cell types to control such movements are not well understood. We report that zebrafish space cadet mutant larvae fail to initiate fast turning movements properly, and we show that this motor phenotype correlates with axonal defects in a small population of commissural hindbrain neurons, which we identify as spiral fiber neurons. Moreover, we demonstrate that severing spiral fiber axons produces space cadet like locomotor defects, thereby providing compelling evidence that the space cadet gene plays an essential role in integrating these neurons into the circuitry that modulates fast turning movements. Finally, we show that axonal defects are restricted to a small set of commissural trajectories, including retinal ganglion cell axons and spiral fiber axons, and that the space cadet gene functions in axonal pathfinding. Together, our results provide a rare example in vertebrates of an individual neuronal cell type that contributes to the expression of a defined motor behavior. Movies available on-line PMID- 11493535 TI - FGF2 promotes skeletogenic differentiation of cranial neural crest cells. AB - The cranial neural crest gives rise to most of the skeletal tissues of the skull. Matrix-mediated tissue interactions have been implicated in the skeletogenic differentiation of crest cells, but little is known of the role that growth factors might play in this process. The discovery that mutations in fibroblast growth factor receptors (FGFRs) cause the major craniosynostosis syndromes implicates FGF-mediated signalling in the skeletogenic differentiation of the cranial neural crest. We now show that, in vitro, mesencephalic neural crest cells respond to exogenous FGF2 in a dose-dependent manner, with 0.1 and 1 ng/ml causing enhanced proliferation, and 10 ng/ml inducing cartilage differentiation. In longer-term cultures, both endochondral and membrane bone are formed. FGFR1, FGFR2 and FGFR3 are all detectable by immunohistochemistry in the mesencephalic region, with particularly intense expression at the apices of the neural folds from which the neural crest arises. FGFRs are also expressed by subpopulations of neural crest cells in culture. Collectively, these findings suggest that FGFs are involved in the skeletogenic differentiation of the cranial neural crest. PMID- 11493536 TI - Hoxa11 and Hoxd11 regulate branching morphogenesis of the ureteric bud in the developing kidney. AB - Hoxa11 and Hoxd11 are functionally redundant during kidney development. Mice with homozygous null mutation of either gene have normal kidneys, but double mutants have rudimentary, or in extreme cases, absent kidneys. We have examined the mechanism for renal growth failure in this mouse model and find defects in ureteric bud branching morphogenesis. The ureteric buds are either unbranched or have an atypical pattern characterized by lack of terminal branches in the midventral renal cortex. The mutant embryos show that Hoxa11 and Hoxd11 control development of a dorsoventral renal axis. By immunohistochemical analysis, Hoxa11 expression is restricted to the early metanephric mesenchyme, which induces ureteric bud formation and branching. It is not found in the ureteric bud. This suggests that the branching defect had been caused by failure of mesenchyme to epithelium signaling. In situ hybridizations with Wnt7b, a marker of the metanephric kidney, show that the branching defect was not simply the result of homeotic transformation of metanephros to mesonephros. Absent Bf2 and Gdnf expression in the midventral mesenchyme, findings that could by themselves account for branching defects, shows that Hoxa11 and Hoxd11 are necessary for normal gene expression in the ventral mesenchyme. Attenuation of normal gene expression along with the absence of a detectable proliferative or apoptotic change in the mutants show that one function of Hoxa11 and Hoxd11 in the developing renal mesenchyme is to regulate differentiation necessary for mesenchymal-epithelial reciprocal inductive interactions. PMID- 11493537 TI - The MCP silencer of the Drosophila Abd-B gene requires both Pleiohomeotic and GAGA factor for the maintenance of repression. AB - Silencing of homeotic gene expression requires the function of cis-regulatory elements known as Polycomb Response Elements (PREs). The MCP silencer element of the Drosophila homeotic gene Abdominal-B has been shown to behave as a PRE and to be required for silencing throughout development. Using deletion analysis and reporter gene assays, we defined a 138 bp sequence within the MCP silencer that is sufficient for silencing of a reporter gene in the imaginal discs. Within the MCP138 fragment, there are four binding sites for the Pleiohomeotic protein (PHO) and two binding sites for the GAGA factor (GAF), encoded by the Trithorax-like gene. PHO and the GAF proteins bind to these sites in vitro. Mutational analysis of PHO and GAF binding sequences indicate that these sites are necessary for silencing in vivo. Moreover, silencing by MCP138 depends on the function of the Trithorax-like gene, and on the function of the PcG genes, including pleiohomeotic. Deletion and mutational analyses show that, individually, either PHO or GAF binding sites retain only weak silencing activity. However, when both PHO and GAF binding sites are present, they achieve strong silencing. We present a model in which robust silencing is achieved by sequential and facilitated binding of PHO and GAF. PMID- 11493538 TI - sprouty4 acts in vivo as a feedback-induced antagonist of FGF signaling in zebrafish. AB - In looking for novel factors involved in the regulation of the fibroblast growth factor (FGF) signaling pathway, we have isolated a zebrafish sprouty4 gene, based on its extensive similarities with the expression patterns of both fgf8 and fgf3. Through gain- and loss-of-function experiments, we demonstrate that Fgf8 and Fgf3 act in vivo to induce the expression of Spry4, which in turn can inhibit activity of these growth factors. When overexpressed at low doses, Spry4 induces loss of cerebellum and reduction in size of the otic vesicle, thereby mimicking the fgf8/acerebellar mutant phenotype. Injections of high doses of Spry4 cause ventralization of the embryo, an opposite phenotype to the dorsalisation induced by overexpression of Fgf8 or Fgf3. Conversely we have shown that inhibition of Spry4 function through injection of antisense morpholino oligonucleotide leads to a weak dorsalization of the embryo, the phenotype expected for an upregulation of Fgf8 or Fgf3 signaling pathway. Finally, we show that Spry4 interferes with FGF signaling downstream of the FGF receptor 1 (FGFR1). In addition, our analysis reveals that signaling through FGFR1/Ras/mitogen-activated protein kinase pathway is involved, not in mesoderm induction, but in the control of the dorsoventral patterning via the regulation of bone morphogenetic protein (BMP) expression. PMID- 11493539 TI - The third wave of myotome colonization by mitotically competent progenitors: regulating the balance between differentiation and proliferation during muscle development. AB - The myotome is formed by a first wave of pioneer cells originating from the entire dorsomedial region of epithelial somites and a second wave that derives from all four lips of the dermomyotome but generates myofibers from only the rostral and caudal edges. Because the precedent progenitors exit the cell cycle upon myotome colonization, subsequent waves must account for consecutive growth. In this study, double labeling with CM-DiI and BrdU revealed the appearance of a third wave of progenitors that enter the myotome as mitotically active cells from both rostral and caudal dermomyotome edges. These cells express the fibroblast growth factor (FGF) receptor FREK and treatment with FGF4 promotes their proliferation and redistribution towards the center of the myotome. Yet, they are negative for MyoD, Myf5 and FGF4, which are, however, expressed in myofibers. The proliferating progenitors first appear around the 30-somite stage in cervical level myotomes and their number continuously increases, making up 85% of total muscle nuclei by embryonic day (E)4. By this stage, generation of second-wave myofibers, which also enter from the extreme lips is still under way. Formation of the latter fibers peaks at 30 somites and progressively decreases with age until E4. Thus, cells in these dermomyotome lips generate simultaneously distinct types of muscle progenitors in changing proportions as a function of age. Consistent with a heterogeneity in the cellular composition of the extreme lips, MyoD is normally expressed in only a subset of these epithelial cells. Treatment with Sonic hedgehog drives most of them to become MyoD positive and then to become myofibers, with a concurrent reduction in the proportion of proliferating muscle precursors. PMID- 11493540 TI - Hindbrain patterning involves graded responses to retinoic acid signalling. AB - Several recent studies have shown that retinoic acid signalling is required for correct patterning of the hindbrain. However, the data from these studies are disparate and the precise role of retinoic acid signalling in patterning the anteroposterior axis of the neural tube remains uncertain. To help clarify this issue, we have cultured a staged series of chick embryos in the presence of an antagonist to the all three retinoic acid receptors. Our data indicate that retinoic acid is the transforming signal involved in the expansion of posterior hindbrain structures. We find that the hindbrain region of the neural tube down to the level of the sixth somite acquires the identity of rhombomere 4 when retinoic acid signalling is blocked. Specification of future rhombomere boundaries has a retinoic acid dependency between stage 5 and stage 10(+) that is lost progressively in an anterior-to-posterior sequence. Furthermore, the application of various concentrations of antagonist shows that successively more posterior rhombomere boundaries require progressively higher concentration of endogenous retinoic acid for their correct positioning, a result that strengthens the hypothesis that a complex retinoid gradient acts to pattern the posterior hindbrain. Our dissection of early retinoic acid functions allows us to re interpret the wide disparity of hindbrain phenotypes previously observed in various models of retinoic acid deficiency. PMID- 11493541 TI - The Drosophila tumor suppressor gene lethal(2)giant larvae is required for the emission of the Decapentaplegic signal. AB - The Drosophila tumor suppressor gene lethal(2) giant larvae (lgl) encodes a cytoskeletal protein required for the change in shape and polarity acquisition of epithelial cells, and also for asymmetric division of neuroblasts. We show here that lgl participates in the emission of Decapentaplegic (Dpp), a member of the transforming growth factor beta (TGFbeta) family, in various developmental processes. During embryogenesis, lgl is required for the dpp-dependent transcriptional activation of zipper (zip), which encodes the non-muscle myosin heavy chain (NMHC), in the dorsalmost ectodermal cells - the leading edge cells. The embryonic expression of known targets of the dpp signaling pathway, such as labial or tinman was abolished or strongly reduced in lgl mutants. lgl mutant cuticles exhibited phenotypes resembling those observed in mutated partners of the dpp signaling pathway. In addition, lgl was required downstream of dpp and upstream of its receptor Thickveins (Tkv) for the dorsoventral patterning of the ectoderm. During larval development, the expression of spalt, a dpp target, was abolished in mutant wing discs, while it was restored by a constitutively activated form of Tkv (Tkv(Q253D)). Taking into account that the activation of dpp expression was unaffected in the mutant, this suggests that lgl function is not required downstream of the Dpp receptor. Finally, the function of lgl responsible for the activation of Spalt expression appeared to be required only in the cells that produce Dpp, and lgl mutant somatic clones behaved non autonomously. We therefore position the activity of lgl in the cells that produce Dpp, and not in those that respond to the Dpp signal. These results are consistent with a same role for lgl in exocytosis and secretion as that proposed for its yeast ortholog sro7/77 and lgl might function in parallel or independently of its well-documented role in the control of epithelial cell polarity. PMID- 11493542 TI - LvNotch signaling plays a dual role in regulating the position of the ectoderm endoderm boundary in the sea urchin embryo. AB - The molecular mechanisms guiding the positioning of the ectoderm-endoderm boundary along the animal-vegetal axis of the sea urchin embryo remain largely unknown. We report here a role for the sea urchin homolog of the Notch receptor, LvNotch, in mediating the position of this boundary. Overexpression of an activated form of LvNotch throughout the embryo shifts the ectoderm-endoderm boundary more animally along the animal-vegetal axis, whereas expression of a dominant negative form shifts the border vegetally. Mosaic experiments that target activated and dominant negative forms of LvNotch into individual blastomeres of the early embryo, combined with lineage analyses, further reveal that LvNotch signaling mediates the position of this boundary by distinct mechanisms within the animal versus vegetal portions of the embryo. In the animal region of the embryo, LvNotch signaling acts cell autonomously to promote endoderm formation more animally, while in the vegetal portion, LvNotch signaling also promotes the ectoderm-endoderm boundary more animally, but through a cell non-autonomous mechanism. We further demonstrate that vegetal LvNotch signaling controls the localization of nuclear beta-catenin at the ectoderm-endoderm boundary. Based on these results, we propose that LvNotch signaling promotes the position of the ectoderm-endoderm boundary more animally via two mechanisms: (1) a cell-autonomous function within the animal region of the embryo, and (2) a cell non-autonomous role in the vegetal region that regulates a signal(s) mediating ectoderm-endoderm position, possibly through the control of nuclear beta-catenin at the boundary. PMID- 11493543 TI - Pre-pattern in the pronephric kidney field of zebrafish. AB - Vertebrate embryos use a series of transient kidneys to regulate fluid balance, osmolarity and metabolic waste during development. The first kidney to form in the embryo is the pronephros. This kidney is composed of several cell types with very different functions and is organized into discrete segments: glomerulus, tubules and nephric duct. The site of origin of these cells is poorly understood, as are their lineage relationships. We have defined regions of the intermediate mesoderm as candidates for the pronephric field by expression patterns of the Wilms' Tumor suppressor gene (wt1), single-minded 1 (sim1) and pax2.1. All of these potential kidney markers are expressed in a stripe of intermediate mesoderm, with distinct, overlapping antero-posterior borders. We labeled small groups of cells in this area by laser uncaging of a fluorescent dextran, and then tracked their fates. We found that there was a bounded contiguous region of the intermediate mesoderm that provides pronephric progenitors. As is true for other organ fields, the pronephric field regulates after focal destruction, such that a normal pronephros forms after laser-mediated removal of the wt1 domain. The progenitors for podocytes, tubular cells and duct are restricted to subdomains within the pronephric field. The most anterior cells in the pronephric field give rise to podocytes. This corresponds to the wt1-expressing region. The next more posterior cells contribute to the tubule, and express both wt1 and pax2.1. The most posterior cells contribute to the nephric duct, and these express pax2.1 and sim1, but not wt1. Thus, there is a field for the pronephric kidney with classical attributes of defined border, pre-pattern and regulation. The pattern of the fate map reflects particular combinations of transcription factors. PMID- 11493544 TI - fringe and Notch specify polar cell fate during Drosophila oogenesis. AB - fringe encodes a glycosyltransferase that modulates the ability of the Notch receptor to be activated by its ligands. We describe studies of fringe function during early stages of Drosophila oogenesis. Animals mutant for hypomorphic alleles of fringe contain follicles with an incorrect number of germline cells, which are separated by abnormally long and disorganized stalks. Analysis of clones of somatic cells mutant for a null allele of fringe localizes the requirement for fringe in follicle formation to the polar cells, and demonstrates that fringe is required for polar cell fate. Clones of cells mutant for Notch also lack polar cells and the requirement for Notch in follicle formation appears to map to the polar cells. Ectopic expression of fringe or of an activated form of Notch can generate an extra polar cell. Our results indicate that fringe plays a key role in positioning Notch activation during early oogenesis, and establish a function for the polar cells in separating germline cysts into individual follicles. PMID- 11493545 TI - Hoxd10 induction and regionalization in the developing lumbosacral spinal cord. AB - We have used Hoxd10 expression as a primary marker of the lumbosacral region to examine the early programming of regional characteristics within the posterior spinal cord of the chick embryo. Hoxd10 is uniquely expressed at a high level in the lumbosacral cord, from the earliest stages of motor column formation through stages of motoneuron axon outgrowth. To define the time period when this gene pattern is determined, we assessed Hoxd10 expression after transposition of lumbosacral and thoracic segments at early neural tube stages. We present evidence that there is an early prepattern for Hoxd10 expression in the lumbosacral neural tube; a prepattern that is established at or before stages of neural tube closure. Cells within more posterior lumbosacral segments have a greater ability to develop high level Hoxd10 expression than the most anterior lumbosacral segments or thoracic segments. During subsequent neural tube stages, this prepattern is amplified and stabilized by environmental signals such that all lumbosacral segments acquire the ability to develop high levels of Hoxd10, independent of their axial environment. Results from experiments in which posterior neural segments and/or paraxial mesoderm segments were placed at different axial levels suggest that signals setting Hoxd10 expression form a decreasing posterior-to-anterior gradient. Our experiments do not, however, implicate adjacent paraxial mesoderm as the only source of graded signals. We suggest, instead, that signals from more posterior embryonic regions influence Hoxd10 expression after the early establishment of a regional prepattern. Concurrent analyses of patterns of LIM proteins and motor column organization after experimental surgeries suggest that the programming of these characteristics follows similar rules. PMID- 11493546 TI - Fate map of early avian cardiac progenitor cells. AB - Cardiogenic fate maps are used to address questions on commitment, differentiation, morphogenesis and organogenesis of the heart. Recently, the accuracy of classical cardiogenic fate maps has been questioned, raising concerns about the conclusions drawn in studies based on these maps. We present accurate fate maps of the heart-forming region (HFR) in avian embryos and show that the putative cardiogenic molecular markers Bmp2 and Nkx2.5 do not govern the boundaries of the HFR as suggested in the literature. Moreover, this paper presents the first fate map of the HFR at stage 4 and addresses a void in the literature concerning rostrocaudal patterning of heart cells between stages 4 and 8. PMID- 11493547 TI - Two distinct domains of Bicoid mediate its transcriptional downregulation by the Torso pathway. AB - The transcriptional activity of the Bicoid morphogen is directly downregulated by the Torso signal transduction cascade at the anterior pole of the Drosophila embryo. This regulation does not involve the homeodomain or direct phosphorylation of Bicoid. We analyse the transcriptional regulation of Bicoid in response to the Torso pathway, using Bicoid variants and fusion proteins between the Bicoid domains and the Gal4 DNA-binding domain. We show that Bicoid possesses three autonomous activation domains. Two of these domains, the serine/threonine rich and the acidic domains, are downregulated by Torso, whereas the third activation domain, which is rich in glutamine, is not. The alanine-rich domain, previously described as an activation domain in vitro, has a repressive activity that is independent of Torso. Thus, Bicoid downregulation by Torso results from a competition between the glutamine-rich domain that is insensitive to Torso and the serine/threonine-rich and acidic activation domains downregulated by Torso. The alanine-rich domain contributes to this process indirectly by reducing the global activity of the protein and in particular the activity of the glutamine rich domain that might otherwise prevent downregulation by Torso. PMID- 11493548 TI - Hierarchical coupling of phytochromes and cryptochromes reconciles stability and light modulation of Arabidopsis development. AB - In plants, development is a continuing process that takes place under strong fluctuations of the light environment. Here we show that in Arabidopsis thaliana plants grown under intense white light, coupling of the photoreceptor cryptochrome 2 to developmental processes is broader than previously appreciated. Compared to the wild type, the cry2 mutant showed reduced activity of a Lhcb1*2 promoter fused to a reporter, and delayed flowering. The cry2 mutation also reduced the inhibition of hypocotyl growth, the unfolding of the cotyledons, the rate of leaf production during the vegetative phase, and the pace of development after transition to the reproductive stage; but these effects were obvious only in the absence of cryptochrome 1 and in some cases phytochrome A and/or phytochrome B. Complementary, the cry2 mutation uncovered novel roles for cryptochrome 1 and phytochrome A. The activity of the Lhcb1*2 promoter was higher in the cry1 cry2 mutant than in the cry2 mutant, suggesting that cry1 could be involved in blue-light repression of photosynthetic genes. Surprisingly, the phyA cry1 cry2 triple mutant flowered earlier and showed better response to photoperiod than the cry1 cry2 double mutant, indicating that phyA is involved in light repression of flowering. Growth and development were severely impaired in the quadruple phyA phyB cry1 cry2 mutant. We propose that stability and light modulation of development are achieved by simultaneous coupling of phytochrome A, phytochrome B, cryptochrome 1 and cryptochrome 2 to developmental processes, in combination with context-dependent hierarchy of their relative activities. PMID- 11493549 TI - Primitive erythropoiesis in the Xenopus embryo: the synergistic role of LMO-2, SCL and GATA-binding proteins. AB - Hematopoietic stem cells are derived from ventral mesoderm during vertebrate development. Gene targeting experiments in the mouse have demonstrated key roles for the basic helix-loop-helix transcription factor SCL and the GATA-binding protein GATA-1 in hematopoiesis. When overexpressed in Xenopus animal cap explants, SCL and GATA-1 are each capable of specifying mesoderm to become blood. Forced expression of either factor in whole embryos, however, does not lead to ectopic blood formation. This apparent paradox between animal cap assays and whole embryo phenotype has led to the hypothesis that additional factors are involved in specifying hematopoietic mesoderm. SCL and GATA-1 interact in a transcriptional complex with the LIM domain protein LMO-2. We have cloned the Xenopus homolog of LMO-2 and show that it is expressed in a similar pattern to SCL during development. LMO-2 can specify hematopoietic mesoderm in animal cap assays. SCL and LMO-2 act synergistically to expand the blood island when overexpressed in whole embryos. Furthermore, co-expression of GATA-1 with SCL and LMO-2 leads to embryos that are ventralized and have blood throughout the dorsal ventral axis. The synergistic effect of SCL, LMO-2 and GATA-1, taken together with the findings that these factors can form a complex in vitro, suggests that this complex specifies mesoderm to become blood during embryogenesis. PMID- 11493550 TI - Spatzle regulates the shape of the Dorsal gradient in the Drosophila embryo. AB - Dorsal-ventral polarity of the Drosophila embryo is established by a nuclear gradient of Dorsal protein, generated by successive gurken-Egfr and spatzle-Toll signaling. Overexpression of extracellular Spatzle dramatically reshapes the Dorsal gradient: the normal single peak is broadened and then refined to two distinct peaks of nuclear Dorsal, to produce two ventral furrows. This partial axis duplication, which mimics the ventralized phenotype caused by reduced gurken Egfr signaling, arises from events in the perivitelline fluid of the embryo and occurs at the level of Spatzle processing or Toll activation. The production of two Dorsal peaks is addressed by a model that invokes action of a diffusible inhibitor, which is proposed to normally regulate the slope of the Dorsal gradient. PMID- 11493551 TI - Fruit development is actively restricted in the absence of fertilization in Arabidopsis. AB - Flowering plants usually require fertilization to form fruit and seed and to initiate floral organ abscission in structures that do not contribute to the fruit. An Arabidopsis mutant that initiates seedless fruit without fertilization (fwf) or parthenocarpy was isolated and characterized to understand the factors regulating the transition between the mature flower and the initiation of seed and fruit development. The fwf mutant is fertile and has normal plant growth and stature. It sets fertile seed following self-pollination and fertilization needs to be prevented to observe parthenocarpy. The initiation of parthenocarpic siliques (fruit) was found to be dependent upon carpel valve identity conferred by FRUITFULL but was independent of the perception of gibberellic acid, shown to stimulate parthenocarpy in Arabidopsis following exogenous application. The recessive nature of fwf is consistent with the involvement of FWF in processes that inhibit fruit growth and differentiation in the absence of fertilization. The enhanced cell division and expansion in the silique mesocarp layer, and increased lateral vascular bundle development imply FWF has roles also in modulating silique growth post-fertilization. Parthenocarpy was inhibited by the presence of other floral organs suggesting that both functional FWF activity and inter-organ communication act in concert to prevent fruit initiation in the absence of fertilization. PMID- 11493552 TI - Physiological rationale for responsiveness of mouse embryonic stem cells to gp130 cytokines. AB - Embryonic stem cells are established directly from the pluripotent epiblast of the preimplantation mouse embryo. Their derivation and propagation are dependent upon cytokine-stimulated activation of gp130 signal transduction. Embryonic stem cells maintain a close resemblance to epiblast in developmental potency and gene expression profile. The presumption of equivalence between embryonic stem cells and epiblast is challenged, however, by the finding that early embryogenesis can proceed in the absence of gp130. To explore this issue further, we have examined the capacity of gp130 mutant embryos to accommodate perturbation of normal developmental progression. Mouse embryos arrest at the late blastocyst stage when implantation is prevented. This process of diapause occurs naturally in lactating females or can be induced experimentally by removal of the ovaries. We report that gp130(-/-) embryos survive unimplanted in the uterus after ovariectomy but, in contrast to wild-type or heterozygous embryos, are subsequently unable to resume development. Inner cell masses explanted from gp130(-/-) delayed blastocysts produce only parietal endoderm, a derivative of the hypoblast. Intact mutant embryos show an absence of epiblast cells, and Hoechst staining and TUNEL analysis reveal a preceding increased incidence of cell death. These findings establish that gp130 signalling is essential for the prolonged maintenance of epiblast in vivo, which is commonly required of mouse embryos in the wild. We propose that the responsiveness of embryonic stem cells to gp130 signalling has its origin in this adaptive physiological function. PMID- 11493553 TI - Hematopoietic regulatory domain of gata1 gene is positively regulated by GATA1 protein in zebrafish embryos. AB - Expression of gata1 is regulated through multiple cis-acting GATA motifs. To elucidate regulatory mechanisms of the gata1 gene, we have used zebrafish. To this end, we isolated and analyzed zebrafish gata1 genomic DNA, which resulted in the discovery of a novel intron that was unknown in previous analyses. This intron corresponds to the first intron of other vertebrate Gata1 genes. GFP reporter analyses revealed that this intron and a distal double GATA motif in the regulatory region are important for the regulation of zebrafish gata1 gene expression. To examine whether GATA1 regulates its own gene expression, we microinjected into embryos a GFP reporter gene linked successively to the gata1 gene regulatory region and to GATA1 mRNA. Surprisingly, ectopic expression of the reporter gene was induced at the site of GATA1 overexpression and was dependent on the distal double GATA motif. Functional domain analyses using transgenic fish lines that harbor the gata1-GFP reporter construct revealed that both the N- and C-terminal zinc-finger domains of GATA1, hence intact GATA1 function, are required for the ectopic GFP expression. These results provide the first in vivo evidence that gata1 gene expression undergoes positive autoregulation. PMID- 11493554 TI - N-terminal fatty-acylation of sonic hedgehog enhances the induction of rodent ventral forebrain neurons. AB - The adult basal ganglia arise from the medial and lateral ganglionic eminences, morphologically distinct structures found in the embryonic telencephalon. We have previously shown that temporal changes in sonic hedgehog (Shh) responsiveness determine the sequential induction of embryonic neurons that populate the medial and lateral ganglionic eminences. In this report, we show that Shh-mediated differentiation of neurons that populate the lateral ganglionic eminence express different combinations of the homeobox-containing transcription factors Dlx, Mash1 and Islet 1/2. Furthermore, we show that N-terminal fatty-acylation of Shh significantly enhances its ability to induce the differentiation of rat E11 telencephalic neurons expressing Dlx, Islet 1/2 or Mash1. Recent evidence indicates that in utero injection of the E9.5 mouse forebrain with retroviruses encoding wild-type Shh induces the ectopic expression of Dlx2 and severe deformities in the brain. In this report, we show that Shh containing a mutation at the site of acylation prevents either of these phenotypes. These results suggest that N-terminal fatty-acylation of Shh may play an important role in Shh dependent signaling during rodent ventral forebrain formation. PMID- 11493555 TI - Proximodistal domain specification and interactions in developing Drosophila appendages. AB - The morphological diversification of appendages represents a crucial aspect of animal body plan evolution. The arthropod antenna and leg are homologous appendages, thought to have arisen via duplication and divergence of an ancestral structure (Snodgrass, R. (1935) Book Principles of Insect Morphology. New York: McGraw-Hill). To gain insight into how variations between the antenna and the leg may have arisen, we have compared the epistatic relationships among three major proximodistal patterning genes, Distal-less, dachshund and homothorax, in the antenna and leg of the insect arthropod Drosophila melanogaster. We find that Drosophila appendages are subdivided into different proximodistal domains specified by specific genes, and that limb-specific interactions between genes and the functions of these genes are crucial for antenna-leg differences. In particular, in the leg, but not in the antenna, mutually antagonistic interactions exist between the proximal and medial domains, as well as between medial and distal domains. The lack of such antagonism in the antenna leads to extensive coexpression of Distal-less and homothorax, which in turn is essential for differentiation of antennal morphology. Furthermore, we report that a fundamental difference between the two appendages is the presence in the leg and absence in the antenna of a functional medial domain specified by dachshund. Our results lead us to propose that the acquisition of particular proximodistal subdomains and the evolution of their interactions has been essential for the diversification of limb morphology. PMID- 11493556 TI - Msx homeobox genes inhibit differentiation through upregulation of cyclin D1. AB - During development, patterning and morphogenesis of tissues are intimately coordinated through control of cellular proliferation and differentiation. We describe a mechanism by which vertebrate Msx homeobox genes inhibit cellular differentiation by regulation of the cell cycle. We show that misexpression of Msx1 via retroviral gene transfer inhibits differentiation of multiple mesenchymal and epithelial progenitor cell types in culture. This activity of Msx1 is associated with its ability to upregulate cyclin D1 expression and Cdk4 activity, while Msx1 has minimal effects on cellular proliferation. Transgenic mice that express Msx1 under the control of the mouse mammary tumor virus long terminal repeat (MMTV LTR) display impaired differentiation of the mammary epithelium during pregnancy, which is accompanied by elevated levels of cyclin D1 expression. We propose that Msx1 gene expression maintains cyclin D1 expression and prevents exit from the cell cycle, thereby inhibiting terminal differentiation of progenitor cells. Our model provides a framework for reconciling the mutant phenotypes of Msx and other homeobox genes with their functions as regulators of cellular proliferation and differentiation during embryogenesis. PMID- 11493557 TI - Analysis of the zebrafish smoothened mutant reveals conserved and divergent functions of hedgehog activity. AB - Despite extensive studies, there are still many unanswered questions regarding the mechanism of hedgehog signaling and the phylogenic conservation of hedgehog function in vertebrates. For example, whether hedgehog signaling in vertebrates requires smoothened is unclear, and the role of hedgehog activity in zebrafish is controversial. We show that inactivation of smoothened by retroviral insertions in zebrafish results in defects that are characteristic of hedgehog deficiencies, including abnormalities in body size, the central nervous system, adaxial mesoderm, cartilage and pectoral fins. We demonstrate that, as in Drosophila, vertebrate smoothened is essential for hedgehog signaling, and functions upstream of protein kinase A. Further analysis of neural tube defects revealed the absence of lateral floor plate and secondary motoneurons, but the presence of medial floor plate and primary motoneurons in smoothened mutant embryos. Blocking maternal hedgehog signaling by cyclopamine eliminates primary motoneurons, but not medial floor plate. Interestingly, even after inhibition of maternal hedgehog activity, the midbrain dopaminergic neurons still form, and looping of the heart does not randomize in the mutants. We also found decreased proliferation and increased apoptosis in the mutants. Taken together, these data demonstrate the conserved role of vertebrate smoothened in the hedgehog signaling pathway, and reveal similarities and differences of hedgehog function between teleosts and amniotes. PMID- 11493558 TI - Haploinsufficiency of the forkhead gene Foxf1, a target for sonic hedgehog signaling, causes lung and foregut malformations. AB - The murine Foxf1 gene, encoding a forkhead - or winged helix - transcription factor, is expressed in splanchnic mesenchyme during organogenesis. The concentration of expression to subepithelial mesenchyme suggested that Foxf1 is activated by paracrine signals from endodermal epithelia. Homozygous Foxf1-null mice die before embryonic day 10, owing to defects in extra-embryonic mesoderm, and do not provide any information about the role of Foxf1 in morphogenesis of endodermally derived organs. We show that, on CD1 genetic background, Foxf1 heterozygote perinatal mortality is around 90%. The haploinsufficiency causes a variable phenotype that includes lung immaturity and hypoplasia, fusion of right lung lobes, narrowing of esophagus and trachea, esophageal atresia and tracheo esophageal fistula. Similar malformations are observed in mutants that are defective in the sonic hedgehog (Shh) signaling pathway, and we show that exogenous Shh activates transcription of Foxf1 in developing lung. Foxf1 mRNA is absent in the lungs, foregut and sclerotomes of Shh(-/-) embryos, but persists in tissues where indian hedgehog (Ihh) is expressed. In lung organ cultures, activation of Foxf1 by Shh is counteracted by bone morphogenetic protein 4 (BMP4). Fibroblast growth factor (FGF) 10 and FGF7 both decrease Foxf1 expression and we speculate that this is mediated by transcriptional activation of epithelial Bmp4 (in the case of FGF10) and by inhibition of Shh expression for FGF7. PMID- 11493559 TI - The homeobox genes vox and vent are redundant repressors of dorsal fates in zebrafish. AB - Ventralizing transcriptional repressors in the Vox/Vent family have been proposed to be important regulators of dorsoventral patterning in the early embryo. While the zebrafish genes vox (vega1) and vent (vega2) both have ventralizing activity in overexpression assays, loss-of-function studies are needed to determine whether these genes have distinct or redundant functions in dorsoventral patterning and to provide critical tests of the proposed regulatory interactions among vox, vent and other genes that act to establish the dorsoventral axis. We show that vox and vent are redundant repressors of dorsal fates in zebrafish. Mutants that lack vox function have little or no dorsoventral patterning defect, and inactivation of either vox or vent by injection of antisense morpholino oligonucleotides has little or no effect on the embryo. In contrast, embryos that lack both vox and vent function have a dorsalized phenotype. Expression of dorsal mesodermal genes, including chordin, goosecoid and bozozok, is strongly expanded in embryos that lack vox and vent function, indicating that the redundant action of vox and vent is required to restrict dorsal genes to their appropriate territories. Our genetic analysis indicates that the dorsalizing transcription factor Bozozok promotes dorsal fates indirectly, by antagonizing the expression of vox and vent. In turn, vox and vent repress chordin expression, restricting its function as an antagonist of ventral fates to the dorsal side of the embryo. Our results support a model in which BMP signaling induces the expression of ventral genes, while vox and vent act redundantly to prevent the expression of chordin, goosecoid and other dorsal genes in the lateral and ventral mesendoderm. PMID- 11493560 TI - Brn3a is a transcriptional regulator of soma size, target field innervation and axon pathfinding of inner ear sensory neurons. AB - The POU domain transcription factors Brn3a, Brn3b and Brn3c are required for the proper development of sensory ganglia, retinal ganglion cells, and inner ear hair cells, respectively. We have investigated the roles of Brn3a in neuronal differentiation and target innervation in the facial-stato-acoustic ganglion. We show that absence of Brn3a results in a substantial reduction in neuronal size, abnormal neuronal migration and downregulation of gene expression, including that of the neurotrophin receptor TrkC, parvalbumin and Brn3b. Selective loss of TrkC neurons in the spiral ganglion of Brn3a(-/-) cochlea leads to an innervation defect similar to that of TrkC(-/-) mice. Most remarkably, our results uncover a novel role for Brn3a in regulating axon pathfinding and target field innervation by spiral and vestibular ganglion neurons. Loss of Brn3a results in severe retardation in development of the axon projections to the cochlea and the posterior vertical canal as early as E13.5. In addition, efferent axons that use the afferent fibers as a scaffold during pathfinding also show severe misrouting. Interestingly, despite the well-established roles of ephrins and EphB receptors in axon pathfinding, expression of these molecules does not appear to be affected in Brn3a(-/-) mice. Thus, Brn3a must control additional downstream genes that are required for axon pathfinding. PMID- 11493561 TI - Emx2 directs the development of diencephalon in cooperation with Otx2. AB - The vertebrate brain is among the most complex biological structures of which the organization remains unclear. Increasing numbers of studies have accumulated on the molecular basis of midbrain/hindbrain development, yet relatively little is known about forebrain organization. Nested expression among Otx and Emx genes has implicated their roles in rostral brain regionalization, but single mutant phenotypes of these genes have not provided sufficient information. In order to genetically determine the interaction between Emx and Otx genes in forebrain development, we have examined Emx2(-/-)Otx2(+/-) double mutants and Emx2 knock-in mutants into the Otx2 locus (Otx2(+/Emx2)). Emx2(-/-)Otx2(+/-) double mutants did not develop diencephalic structures such as ventral thalamus, dorsal thalamus/epithalamus and anterior pretectum. The defects were attributed to the loss of the Emx2-positive region at the three- to four-somite stage, when its expression occurs in the laterocaudal forebrain primordia. Ventral structures such as the hypothalamus, mammillary region and tegmentum developed normally. Moreover, dorsally the posterior pretectum and posterior commissure were also present in the double mutants. In contrast, Otx2(+/Emx2) knock-in mutants displayed the majority of these diencephalic structures; however, the posterior pretectum and posterior commissure were specifically absent. Consequently, development of the dorsal and ventral thalamus and anterior pretectum requires cooperation between Emx2 and Otx2, whereas Emx2 expression is incompatible with development of the commissural region of the pretectum. PMID- 11493562 TI - Evidence for a role of protein kinase C in FGF signal transduction in the developing chick limb bud. AB - In developing limbs, numerous signaling molecules have been identified but less is known about the mechanisms by which such signals direct patterning. We have explored signal transduction pathways in the chicken limb bud. A cDNA encoding RACK1, a protein that binds and stabilizes activated protein kinase C (PKC), was isolated in a screen for genes induced by retinoic acid (RA) in the chick wing bud. Fibroblast growth factor (FGF) also induced RACK1 and such induction of RACK1 expression was accompanied by a significant augmentation in the number of active PKC molecules and an elevation of PKC enzymatic activity. This suggests that PKCs mediate signal transduction in the limb bud. Application of chelerythrine, a potent PKC inhibitor, to the presumptive wing region resulted in buds that did not express sonic hedgehog (Shh) and developed into wings that were severely truncated. This observation suggests that the expression of Shh depends on PKCs. Providing ectopic SHH protein, RA or ZPA grafts overcome the effects of blocking PKC with chelerythrine and resulted in a rescue of the wing morphology. Taken together, these findings suggest that the responsiveness of Shh to FGF is mediated, at least in part, by PKCs. PMID- 11493563 TI - Inductive signal and tissue responsiveness defining the tectum and the cerebellum. AB - The mes/metencephalic boundary (isthmus) has an organizing activity for mesencephalon and metencephalon. The candidate signaling molecule is Fgf8 whose mRNA is localized in the region where the cerebellum differentiates. Responding to this signal, the cerebellum differentiates in the metencephalon and the tectum differentiates in the mesencephalon. Based on the assumption that strong Fgf8 signal induces the cerebellum and that the Fgf8b signal is stronger than that of Fgf8a, we carried out experiments to misexpress Fgf8b and Fgf8a in chick embryos. Fgf8a did not affect the expression pattern of Otx2, Gbx2 or Irx2. En2 expression was upregulated in the mesencephalon and in the diencephalon by Fgf8a. Consequently, Fgf8a misexpression resulted in the transformation of the presumptive diencephalon to the fate of the mesencephalon. In contrast, Fgf8b repressed Otx2 expression, but upregulated Gbx2 and Irx2 expression in the mesencephalon. As a result, Fgf8b completely changed the fate of the mesencephalic alar plate to cerebellum. Quantitative analysis showed that Fgf8b signal is 100 times stronger than Fgf8a signal. Co-transfection of Fgf8b with Otx2 indicates that Otx2 is a key molecule in mesencephalic generation. We have shown by RT-PCR that both Fgf8a and Fgf8b are expressed, Fgf8b expression prevailing in the isthmic region. The results all support our working hypothesis that the strong Fgf8 signal induces the neural tissue around the isthmus to differentiate into the cerebellum. PMID- 11493564 TI - Consequences of Hox gene duplication in the vertebrates: an investigation of the zebrafish Hox paralogue group 1 genes. AB - As a result of a whole genome duplication event in the lineage leading to teleosts, the zebrafish has seven clusters of Hox patterning genes, rather than four, as described for tetrapod vertebrates. To investigate the consequences of this genome duplication, we have carried out a detailed comparison of genes from a single Hox paralogue group, paralogue group (PG) 1. We have analyzed the sequences, expression patterns and potential functions of all four of the zebrafish PG1 Hox genes, and compared our data with that available for the three mouse genes. As the basic functions of Hox genes appear to be tightly constrained, comparison with mouse data has allowed us to identify specific changes in the developmental roles of Hox genes that have occurred during vertebrate evolution. We have found variation in expression patterns, amino acid sequences within functional domains, and potential gene functions both within the PG1 genes of zebrafish, and in comparison to mouse PG1 genes. We observed novel expression patterns in the midbrain, such that zebrafish hoxa1a and hoxc1a are expressed anterior to the domain traditionally thought to be under Hox patterning control. The hoxc1a gene shows significant coding sequence changes in known functional domains, which correlate with a reduced capacity to cause posteriorizing transformations. Moreover, the hoxb1 duplicate genes have differing functional capacities, suggesting divergence after duplication. We also find that an intriguing function 'shuffling' between paralogues has occurred, such that one of the zebrafish hoxb1 duplicates, hoxb1b, performs the role in hindbrain patterning played in mouse by the non-orthologous Hoxa1 gene. PMID- 11493565 TI - Knockout mice reveal a contribution of the extracellular matrix molecule tenascin C to neural precursor proliferation and migration. AB - The extracellular matrix glycoprotein tenascin-C is widely expressed in the vertebrate central nervous system (CNS) during development and repair. Despite multiple effects of tenascin-C on cell behaviour in culture, no structural abnormalities of the CNS and other organs have been found in adult tenascin-C null mice, raising the question of whether this glycoprotein has a significant role in vivo. Using a transgenic approach, we have demonstrated that tenascin-C regulates both cell proliferation and migration in oligodendrocyte precursors during development. Knockout mice show increased rates of oligodendrocyte precursor migration along the optic nerve and reduced rates of oligodendrocyte precursor proliferation in different regions of the CNS. Levels of programmed cell death were reduced in areas of myelination at later developmental stages, providing a potential corrective mechanism for any reduction in cell numbers that resulted from the proliferation phenotype. The effects on cell proliferation are mediated via the alphavbeta3 integrin and an interaction with the platelet derived growth factor-stimulated mitogenic pathway, emphasising the importance of both CNS extracellular matrix and integrin growth factor interactions in the regulation of neural precursor behaviour. PMID- 11493567 TI - The role of an endogenous PKA inhibitor, PKIalpha, in organizing left-right axis formation. AB - Protein kinase inhibitor (PKI) is an endogenous inhibitor of cAMP-dependent protein kinase A (PKA). We have found that the alpha-isoform of PKI (PKIalpha) is asymmetrically expressed along the left-right (L-R) axis in chick embryos. At stage 6, PKIalpha is expressed on the right side of the node, and this asymmetric expression continues until stage 7+. After stage 8, PKIalpha expression returns symmetric. Treatment of embryos with antisense PKIalpha oligonucleotides increased the incidence of reversed heart looping. Antisense oligonucleotides also induced ectopic expression of the left-specific genes Nodal and Pitx2, and suppressed the expression of the right-specific gene SnR in the right lateral plate mesoderm. Similarly, treatment with PKA activators forskolin and Sp-cAMPs resulted in both reversed heart looping and bilateral expression of NODAL: Ectopic activin induced PKIalpha on the left side of the node, while ectopic Shh and anti-Shh antibody had no effect on PKIalpha expression. Taken together, these data suggest that PKIalpha induced by an activin-like molecule, through the inhibition of PKA activity, suppresses the Nodal-Pitx2 pathway on the right side of the body. PMID- 11493568 TI - A new approach reveals syncytia within the visceral musculature of Drosophila melanogaster. AB - In order to reveal syncytia within the visceral musculature of Drosophila melanogaster, we have combined the GAL4/UAS system with the single-cell transplantation technique. After transplantation of single cells from UAS-GFP donor embryos into ubiquitously GAL4-expressing recipients, the expression of the reporter gene was exclusively activated in syncytia containing both donor- and recipient-derived nuclei. In the first trial, we tested the system in the larval somatic musculature, which is already known to consist of syncytia. By this means we could show that most of the larval somatic muscles are generated by clonally non-related cells. Moreover, using this approach we were able to detect syncytia within the visceral musculature - a tissue that has previously been described as consisting of mononuclear cells. Both the longitudinal visceral musculature of the midgut and the circular musculature of the hindgut consist of syncytia and persist through metamorphosis. This novel application of the transplantation technique might be a powerful tool to trace syncytia in any organism using the GAL4/UAS system. PMID- 11493566 TI - Math5 is required for retinal ganglion cell and optic nerve formation. AB - The vertebrate retina contains seven major neuronal and glial cell types in an interconnected network that collects, processes and sends visual signals through the optic nerve to the brain. Retinal neuron differentiation is thought to require both intrinsic and extrinsic factors, yet few intrinsic gene products have been identified that direct this process. Math5 (Atoh7) encodes a basic helix-loop-helix (bHLH) transcription factor that is specifically expressed by mouse retinal progenitors. Math5 is highly homologous to atonal, which is critically required for R8 neuron formation during Drosophila eye development. Like R8 cells in the fly eye, retinal ganglion cells (RGCs) are the first neurons in the vertebrate eye. Here we show that Math5 mutant mice are fully viable, yet lack RGCs and optic nerves. Thus, two evolutionarily diverse eye types require atonal gene family function for the earliest stages of retinal neuron formation. At the same time, the abundance of cone photoreceptors is significantly increased in Math5(-/-) retinae, suggesting a binary change in cell fate from RGCs to cones. A small number of nascent RGCs are detected during embryogenesis, but these fail to develop further, suggesting that committed RGCs may also require Math5 function. PMID- 11493569 TI - Requirement of FoxD3-class signaling for neural crest determination in Xenopus. AB - Fox factors (winged-helix transcription factors) play important roles in early embryonic patterning. We show here that FoxD3 (Forkhead 6) regulates neural crest determination in Xenopus embryos. Expression of FoxD3 in the presumptive neural crest region starts at the late gastrula stage in a manner similar to that of Slug, and overlaps with that of Zic-r1. When overexpressed in the embryo and in ectodermal explants, FoxD3 induces expression of neural crest markers. Attenuation of FoxD3-related signaling by a dominant-negative FoxD3 construct (FoxD3delN) inhibits neural crest differentiation in vivo without suppressing the CNS marker Sox2. Interestingly, these loss-of-function phenotypes are reversed by coinjecting SLUG: In animal cap explants, neural crest differentiation induced by Slug and Wnt3a is also inhibited by FoxD3delN but not by a dominant-negative form of XBF2. Loss-of-function studies using dominant-negative forms of FoxD3 and Slug indicate that Slug induction by Zic factors requires FoxD3-related signaling, and that FoxD3 and Slug have different requirements in inducing downstream neural crest markers. These data demonstrate that FoxD3 (or its closely related factor) is an essential upstream regulator of neural crest determination. PMID- 11493570 TI - Patterning of fast and slow fibers within embryonic muscles is established independently of signals from the surrounding mesenchyme. AB - During embryonic development, and before functional innervation, a highly stereotypic pattern of slow- and fast-contracting primary muscle fibers is established within individual muscles of the limbs, from distinct populations of myoblasts. A difference between the fiber-type pattern found within chicken and quail pectoral muscles was exploited to investigate the contributions of somite derived myogenic precursors and lateral plate-derived mesenchymal stroma to the establishment of muscle fiber-type patterns. Chimeric chicken/quail embryos were constructed by reciprocal transplantation of somites or lateral plate mesoderm at stages prior to muscle formation. Muscle fibers derived from quail myogenic precursors that had migrated into chicken stroma showed a quail pattern of mixed fast- and slow-contracting muscle fibers. Conversely, chicken myogenic precursors that had migrated into quail stroma showed a chicken pattern of nearly exclusive fast muscle fiber formation. These results demonstrate in vivo an intrinsic commitment to fiber-type on the part of the myoblast, independent of extrinsic signals it receives from the mesenchymal stroma in which it differentiates. PMID- 11493571 TI - Hedgehog-dependent oligodendrocyte lineage specification in the telencephalon. AB - In the caudal neural tube, oligodendrocyte progenitors (OLPs) originate in the ventral neuroepithelium under the influence of Sonic hedgehog (SHH), then migrate throughout the spinal cord and brainstem before differentiating into myelin forming cells. We present evidence that oligodendrogenesis in the anterior neural tube follows a similar pattern. We show that OLPs in the embryonic mouse forebrain express platelet-derived growth factor alpha-receptors (PDGFRA), as they do in more caudal regions. They first appear within a region of anterior hypothalamic neuroepithelium that co-expresses mRNA encoding SHH, its receptor PTC1 (PTCH) and the transcription factors OLIG1, OLIG2 and SOX10. Pdgfra-positive progenitors later spread through the forebrain into areas where Shh is not expressed, including the cerebral cortex. Cyclopamine inhibited OLP development in cultures of mouse basal forebrain, suggesting that hedgehog (HH) signalling is obligatory for oligodendrogenesis in the ventral telencephalon. Moreover, Pdgfra positive progenitors did not appear on schedule in the ventral forebrains of Nkx2.1 null mice, which lack the telencephalic domain of Shh expression. However, OLPs did develop in cultures of Nkx2.1(-/-) basal forebrain and this was blocked by cyclopamine. OLPs also developed in neocortical cultures, even though Shh transcripts could not be detected in the embryonic cortex. Here, too, the appearance of OLPs was suppressed by cyclopamine. In keeping with these findings, we detected mRNA encoding SHH and Indian hedgehog (IHH) in both Nkx2.1(-/-) basal forebrain cultures and neocortical cultures. Overall, the data are consistent with the idea that OLPs in the telencephalon, possibly even some of those in the cortex, develop under the influence of SHH in the ventral forebrain. PMID- 11493573 TI - CRTF is a novel transcription factor that regulates multiple stages of Dictyostelium development. AB - During aggregation, Dictyostelium establish nanomolar oscillation waves of extracellular cAMP, but as development progresses, cells become responsive to higher, non-fluctuating concentrations of cAMP. The regulation of the promoter responsible for expression of cAMP receptor subtype 1, CAR1, during aggregation reflects these signaling variations. Transcription of CAR1 from the early, aggregation promoter is activated by cAMP pulsing, but is repressed by continuous exposure to micromolar concentrations of cAMP. Deletion and mutation analyses of this promoter had defined an element essential for cAMP-regulated expression, and mobility shift assay, DNA crosslinking and DNase I footprinting experiments had identified a nuclear protein (CRTF) with zinc-dependent sequence binding specificity. In our study, CRTF was purified to homogeneity, peptides were sequenced and full-length cDNAs were obtained. The deduced CRTF protein is approximately 100 kDa with a C-terminal, zinc finger-like motif required for DNA binding; CRTF purified from cells, however, represents only a 40 kDa C-terminal fragment that retains DNA-binding activity. As might have been predicted if CRTF were essential for the regulation of CAR1, crtf-null strains fail to develop under standard conditions or to exhibit induced expression of CAR1 or other cAMP regulated genes. Furthermore, crtf-nulls also fail to sporulate, even under conditions that bypass the dependence on early cAMP signaling pathways. In addition, early developmental events of crtf-null strains could be rescued with exogenous cAMP treatment, constitutive expression of CAR1 or co-development with wild-type cells; however, these treatments were insufficient to promote sporulation. This suggests a cell-autonomous role for CRTF during late development that is separate from its capacity to control CAR1 expression. Finally, ablation of CRTF promotes a precocious induction of certain cAMP dependent gene expression pathways. We suggest that CRTF may function to help insulate distinct pathways from simultaneous and universal activation by cAMP. CRTF, thus, exhibits multiple complex and independent regulatory functions during Dictyostelium development. PMID- 11493572 TI - Large-scale cDNA analysis of the maternal genetic information in the egg of Halocynthia roretzi for a gene expression catalog of ascidian development. AB - The ascidian egg is a well-known mosaic egg. In order to investigate the molecular nature of the maternal genetic information stored in the egg, we have prepared cDNAs from the mRNAs in the fertilized eggs of the ascidian, Halocynthia roretzi. The cDNAs of the ascidian embryo were sequenced, and the localization of individual mRNA was examined in staged embryos by whole-mount in situ hybridization. The data obtained were stored in the database MAGEST (http://www.genome.ad.jp/magest) and further analyzed. A total of 4240 cDNA clones were found to represent 2221 gene transcripts, including at least 934 different protein-coding sequences. The mRNA population of the egg consisted of a low prevalence, high complexity sequence set. The majority of the clones were of the rare sequence class, and of these, 42% of the clones showed significant matches with known peptides, mainly consisting of proteins with housekeeping functions such as metabolism and cell division. In addition, we found cDNAs encoding components involved in different signal transduction pathways and cDNAs encoding nucleotide-binding proteins. Large-scale analyses of the distribution of the RNA corresponding to each cDNA in the eight-cell, 110-cell and early tailbud embryos were simultaneously carried out. These analyses revealed that a small fraction of the maternal RNAs were localized in the eight-cell embryo, and that 7.9% of the clones were exclusively maternal, while 40.6% of the maternal clones showed expression in the later stages. This study provides global insights about the genes expressed during early development. PMID- 11493574 TI - Xenopus Dishevelled signaling regulates both neural and mesodermal convergent extension: parallel forces elongating the body axis. AB - During amphibian development, non-canonical Wnt signals regulate the polarity of intercalating dorsal mesoderm cells during convergent extension. Cells of the overlying posterior neural ectoderm engage in similar morphogenetic cell movements. Important differences have been discerned in the cell behaviors associated with neural and mesodermal cell intercalation, raising the possibility that different mechanisms may control intercalations in these two tissues. In this report, targeted expression of mutants of Xenopus Dishevelled (Xdsh) to neural or mesodermal tissues elicited different defects that were consistent with inhibition of either neural or mesodermal convergent extension. Expression of mutant Xdsh also inhibited elongation of neural tissues in vitro in Keller sandwich explants and in vivo in neural plate grafts. Targeted expression of other Wnt signaling antagonists also inhibited neural convergent extension in whole embryos. In situ hybridization indicated that these defects were not due to changes in cell fate. Examination of embryonic phenotypes after inhibition of convergent extension in different tissues reveals a primary role for mesodermal convergent extension in axial elongation, and a role for neural convergent extension as an equalizing force to produce a straight axis. This study demonstrates that non-canonical Wnt signaling is a common mechanism controlling convergent extension in two very different tissues in the Xenopus embryo and may reflect a general conservation of control mechanisms in vertebrate convergent extension. PMID- 11493575 TI - A UAS site substitution approach to the in vivo dissection of promoters: interplay between the GATAb activator and the AEF-1 repressor at a Drosophila ecdysone response unit. AB - An ecdysone response unit (EcRU) directs the expression of the Fat body protein 1 (Fbp1) gene in the third instar larval Drosophila fat body. The tissue-specific activity of this regulatory element necessitates the binding of both the ligand activated EcR/USP ecdysone receptor and GATAb. To analyze the role played by GATAb in the regulation of the Fbp1 EcRU activity, we have replaced the GATA binding sites GBS1, GBS2 and GBS3 in the Fbp1 EcRU with UAS sites for the yeast GAL4 activator and tested the activity of the mutagenized Fbp1 EcRUs in transgenic lines, either in the presence or absence of ubiquitously expressed GAL4. Our results reveal that GATAb plays two distinguishable roles at the Fbp1 EcRU that contribute to the tissue-specific activity of this regulatory element. On the one hand, GATAb mediates a fat body-specific transcriptional activation. On the other hand, it antagonizes specifically in the fat body a ubiquitous repressor that maintains the Fbp1 EcRU in an inactive state, refractory to activation by GAL4. We identified this repressor as AEF-1, a factor previously shown to be involved in the regulation of the Drosophila Adh and yp1-yp2 genes. These results show that, for a functional dissection of complex promoter dependent regulatory pathways, the replacement of specific regulatory target sites by UAS GAL4 binding sites is a powerful alternative to the widely used disruption approach. PMID- 11493577 TI - A large-scale analysis of mRNAs expressed by primary mesenchyme cells of the sea urchin embryo. AB - The primary mesenchyme cells (PMCs) of the sea urchin embryo have been an important model system for the analysis of cell behavior during gastrulation. To gain an improved understanding of the molecular basis of PMC behavior, a set of 8293 expressed sequenced tags (ESTs) was derived from an enriched population of mid-gastrula stage PMCs. These ESTs represented approximately 1200 distinct proteins, or about 15% of the mRNAs expressed by the gastrula stage embryo. 655 proteins were similar (P<10(-7) by BLAST comparisons) to other proteins in GenBank, for which some information is available concerning expression and/or function. Another 116 were similar to ESTs identified in other organisms, but not further characterized. We conservatively estimate that sequences encoding at least 435 additional proteins were included in the pool of ESTs that did not yield matches by BLAST analysis. The collection of newly identified proteins includes many candidate regulators of primary mesenchyme morphogenesis, including PMC-specific extracellular matrix proteins, cell surface proteins, spicule matrix proteins and transcription factors. This work provides a basis for linking specific molecular changes to specific cell behaviors during gastrulation. Our analysis has also led to the cloning of several key components of signaling pathways that play crucial roles in early sea urchin development. PMID- 11493576 TI - Extranuclear sequestration of phospho-Jun N-terminal kinase and distorted villi produced by activated Rac1 in the intestinal epithelium of chimeric mice. AB - Previously, we used a genetic mosaic system to conduct an in vivo analysis of the effects of Rac1 activation on the developing intestinal epithelium ( Stappenbeck, T. S. and Gordon, J. I. (2000) Development 127, 2629-2642). Expression of a constitutively active human Rac1 (Rac1Leu61) in the 129/Sv-derived small intestinal epithelium of C57Bl/6-ROSA26<-->129/Sv chimeric mice led to precocious differentiation of some lineages with accompanying alterations in their apical actin. We have now explored the underlying mechanisms. Rac1Leu61 leads to accumulation of the 46 kDa form of phosphorylated Jun N-terminal kinase (p-Jnk) in the apical cytoplasm, but not in the nucleus of E18.5 proliferating and differentiating intestinal epithelial cells. The effect is cell-autonomous, selective for this mitogen-activated protein kinase family member, and accompanied by apical cytoplasmic accumulation of p21-activated kinase. c-Jun, a downstream nuclear target of p-Jnk, does not show evidence of enhanced phosphorylation, providing functional evidence for cytoplasmic sequestration of p Jnk in Rac1Leu61-expressing epithelium. In adult chimeras, Rac1 activation augments cell proliferation in crypts of Lieberkuhn, without a compensatory change in basal apoptosis and produces a dramatic, very unusual widening of villi. These results reveal a novel in vivo paradigm for Rac1 activation involving p-Jnk-mediated signaling at a distinctive extra-nuclear site, with associated alterations in the actin cytoskeleton. They also provide a new perspective about the determinants of small intestinal villus morphogenesis. PMID- 11493578 TI - Optimizing gh therapy in adults and children. AB - Until the advent of modern neuroradiological imaging techniques in 1989, a diagnosis of GH deficiency in adults carried little significance other than as a marker of hypothalamo-pituitary disease. The relatively recent recognition of a characteristic clinical syndrome associated with failure of spontaneous GH secretion and the potential reversal of many of its features with recombinant human GH has prompted a closer examination of the physiological role of GH after linear growth is complete. The safe clinical practice of GH replacement demands a method of judging overall GH status, but there is no biological marker in adults that is the equivalent of linear growth in a child by which to judge the efficacy of GH replacement. Assessment of optimal GH replacement is made difficult by the apparent diverse actions of GH in health, concern about the avoidance of iatrogenic acromegaly, and the growing realization that an individual's risk of developing certain cancers may, at least in part, be influenced by cumulative exposure to the chief mediator of GH action, IGF-I. As in all areas of clinical practice, strategies and protocols vary between centers, but most physicians experienced in the management of pituitary disease agree that GH is most appropriately begun at low doses, building up slowly to the final maintenance dose. This, in turn, is best determined by a combination of clinical response and measurement of serum IGF-I, avoiding supraphysiological levels of this GH dependent peptide. Numerous studies have helped define the optimum management of GH replacement during childhood. The recent requirement to measure and monitor GH status in adult life has called into question the appropriateness of simplistic weight- and surface area-based dosing regimens for the management of GH deficiency in childhood, with reliance on linear growth as the sole marker of GH action. It is clear that the monitoring of parameters other than linear growth to help refine GH therapy should now be incorporated into childhood GH treatment protocols. Further research will be required to define the optimal management of the transition from pediatric to adult GH replacement; this transition will only be possible once the benefits of GH in mature adults are defined and accepted by pediatric and adult endocrinologists alike. PMID- 11493579 TI - Plasma membrane transport of thyroid hormones and its role in thyroid hormone metabolism and bioavailability. AB - Although it was originally believed that thyroid hormones enter target cells by passive diffusion, it is now clear that cellular uptake is effected by carrier mediated processes. Two stereospecific binding sites for each T4 and T3 have been detected in cell membranes and on intact cells from humans and other species. The apparent Michaelis-Menten values of the high-affinity, low-capacity binding sites for T4 and T3 are in the nanomolar range, whereas the apparent Michaelis- Menten values of the low-affinity, high-capacity binding sites are usually in the lower micromolar range. Cellular uptake of T4 and T3 by the high-affinity sites is energy, temperature, and often Na+ dependent and represents the translocation of thyroid hormone over the plasma membrane. Uptake by the low-affinity sites is not dependent on energy, temperature, and Na+ and represents binding of thyroid hormone to proteins associated with the plasma membrane. In rat erythrocytes and hepatocytes, T3 plasma membrane carriers have been tentatively identified as proteins with apparent molecular masses of 52 and 55 kDa. In different cells, such as rat erythrocytes, pituitary cells, astrocytes, and mouse neuroblastoma cells, uptake of T4 and T3 appears to be mediated largely by system L or T amino acid transporters. Efflux of T3 from different cell types is saturable, but saturable efflux of T4 has not yet been demonstrated. Saturable uptake of T4 and T3 in the brain occurs both via the blood-brain barrier and the choroid plexus cerebrospinal fluid barrier. Thyroid hormone uptake in the intact rat and human liver is ATP dependent and rate limiting for subsequent iodothyronine metabolism. In starvation and nonthyroidal illness in man, T4 uptake in the liver is decreased, resulting in lowered plasma T3 production. Inhibition of liver T4 uptake in these conditions is explained by liver ATP depletion and increased concentrations of circulating inhibitors, such as 3-carboxy-4-methyl-5-propyl-2 furanpropanoic acid, indoxyl sulfate, nonesterified fatty acids, and bilirubin. Recently, several organic anion transporters and L type amino acid transporters have been shown to facilitate plasma membrane transport of thyroid hormone. Future research should be directed to elucidate which of these and possible other transporters are of physiological significance, and how they are regulated at the molecular level. PMID- 11493580 TI - Vitamin D deficiency and secondary hyperparathyroidism in the elderly: consequences for bone loss and fractures and therapeutic implications. AB - Vitamin D deficiency is common in the elderly, especially in the housebound and in geriatric patients. The establishment of strict diagnostic criteria is hampered by differences in assay methods for 25-hydroxyvitamin D. The synthesis of vitamin D3 in the skin under influence of UV light decreases with aging due to insufficient sunlight exposure, and a decreased functional capacity of the skin. The diet contains a minor part of the vitamin D requirement. Vitamin D deficiency in the elderly is less common in the United States than elsewhere due to the fortification of milk and use of supplements. Deficiency in vitamin D causes secondary hyperparathyroidism, high bone turnover, bone loss, mineralization defects, and hip and other fractures. Less certain consequences include myopathy and falls. A diet low in calcium may cause an increased turnover of vitamin D metabolites and thereby aggravate vitamin D deficiency. Prevention is feasible by UV light exposure, food fortification, and supplements. Vitamin D3 supplementation causes a decrease of the serum PTH concentration, a decrease of bone turnover, and an increase of bone mineral density. Vitamin D3 and calcium may decrease the incidence of hip and other peripheral fractures in nursing home residents. Vitamin D3 is recommended in housebound elderly, and it may be cost effective in hip fracture prevention in selected risk groups. PMID- 11493581 TI - Stressor specificity of central neuroendocrine responses: implications for stress related disorders. AB - Despite the fact that many research articles have been written about stress and stress-related diseases, no scientifically accepted definition of stress exists. Selye introduced and popularized stress as a medical and scientific idea. He did not deny the existence of stressor-specific response patterns; however, he emphasized that such responses did not constitute stress, only the shared nonspecific component. In this review we focus mainly on the similarities and differences between the neuroendocrine responses (especially the sympathoadrenal and the sympathoneuronal systems and the hypothalamo-pituitary-adrenocortical axis) among various stressors and a strategy for testing Selye's doctrine of nonspecificity. In our experiments, we used five different stressors: immobilization, hemorrhage, cold exposure, pain, or hypoglycemia. With the exception of immobilization stress, these stressors also differed in their intensities. Our results showed marked heterogeneity of neuroendocrine responses to various stressors and that each stressor has a neurochemical "signature." By examining changes of Fos immunoreactivity in various brain regions upon exposure to different stressors, we also attempted to map central stressor-specific neuroendocrine pathways. We believe the existence of stressor-specific pathways and circuits is a clear step forward in the study of the pathogenesis of stress related disorders and their proper treatment. Finally, we define stress as a state of threatened homeostasis (physical or perceived treat to homeostasis). During stress, an adaptive compensatory specific response of the organism is activated to sustain homeostasis. The adaptive response reflects the activation of specific central circuits and is genetically and constitutionally programmed and constantly modulated by environmental factors. PMID- 11493583 TI - The Endocrine Society Awards. PMID- 11493584 TI - The Endocrine Society 2001 annual awards. PMID- 11493585 TI - Decision or discrimination; the female deficit in the life sciences. PMID- 11493586 TI - On research in a small country. Basic research and its practical applications. PMID- 11493587 TI - Of maize and men. Is the endorsement of GM crops science or politics? PMID- 11493588 TI - Bioscience and its impact on developing countries. A view from Thailand. PMID- 11493589 TI - Losing them is not an option. Women in higher positions in the natural sciences are few and far between. As society cannot afford to lose them and the investment in their education, the question is, what to do about it? PMID- 11493590 TI - Searching for discrimination. Are women treated fairly in the EMBO postdoctoral fellowship scheme? PMID- 11493591 TI - Body, heal thyself. Research on stem cells and therapeutic cloning: great hope tinged with caution. PMID- 11493592 TI - New twists on embryonic patterning. EMBO workshop: embryonic organizer signaling: the next frontiers. PMID- 11493593 TI - A novel lectin in the secretory pathway. An elegant mechanism for glycoprotein elimination. PMID- 11493594 TI - Chromosome motors on the move. From motion to spindle checkpoint activity. AB - Spindle assembly and chromosome segregation require the concerted activities of a variety of microtubule-dependent motors. This review focuses on our current knowledge of the roles played by the chromosome-associated motors during mitosis. While some appear to function conventionally in moving chromosomes along microtubules others seem to act in different ways. For example, by docking microtubules to chromosome arms, chromatin-associated motors prevent chromosome loss and participate in spindle formation and stability. Kinetochore motors participate in the formation of stable kinetochore fibers or in the control of microtubule dynamics and are involved in spindle checkpoint activity. Chromosome associated motors thus appear to be key molecules that function in complementary ways to ensure the accuracy of chromosome segregation. PMID- 11493595 TI - Cavities and packing defects in the structural dynamics of myoglobin. AB - Small globular proteins contain internal cavities and packing defects that reduce thermodynamic stability but seem to play a role in controlling function by defining pathways for the diffusion of the ligand/substrate to the active site. In the case of myoglobin (Mb), a prototype for structure-function relationship studies, the photosensitivity of the adduct of the reduced protein with CO, O2 and NO allows events related to the migration of the ligand through the matrix to be followed. The crystal structures of intermediate states of wild-type (wt) and mutant Mbs show the photolysed CO to be located either in the distal heme pocket (primary docking site) or in one of two alternative cavities (secondary docking sites) corresponding to packing defects accessible to an atom of xenon. These results convey the general picture that pre-existing internal cavities are involved in controlling the dynamics and reactivity of the reactions of Mb with O2 and other ligands, including NO. PMID- 11493596 TI - Multiple pathways contribute to nuclear import of core histones. AB - Nuclear import of the four core histones H2A, H2B, H3 and H4 is one of the main nuclear import activities during S-phase of the cell cycle. However, the molecular machinery facilitating nuclear import of core histones has not been elucidated. Here, we investigated the pathways by which histone import can occur. First, we show that core histone import can be competed by the BIB (beta-like import receptor binding) domain of ribosomal protein L23a suggesting that histone import is an importin mediated process. Secondly, affinity chromatography on immobilized core histones revealed that several members of the importin beta family of transport receptors are able to interact with core histones. Finally, we demonstrate that at least four known and one novel importin, importin 9, can mediate nuclear import of core histones into the nuclei of permeabilized cells. Our results suggest that multiple pathways of import exist to provide efficient nuclear uptake of these abundant, essential proteins. PMID- 11493597 TI - In silico identification of novel selenoproteins in the Drosophila melanogaster genome. AB - In selenoproteins, incorporation of the amino acid selenocysteine is specified by the UGA codon, usually a stop signal. The alternative decoding of UGA is conferred by an mRNA structure, the SECIS element, located in the 3'-untranslated region of the selenoprotein mRNA. Because of the non-standard use of the UGA codon, current computational gene prediction methods are unable to identify selenoproteins in the sequence of the eukaryotic genomes. Here we describe a method to predict selenoproteins in genomic sequences, which relies on the prediction of SECIS elements in coordination with the prediction of genes in which the strong codon bias characteristic of protein coding regions extends beyond a TGA codon interrupting the open reading frame. We applied the method to the Drosophila melanogaster genome, and predicted four potential selenoprotein genes. One of them belongs to a known family of selenoproteins, and we have tested experimentally two other predictions with positive results. Finally, we have characterized the expression pattern of these two novel selenoprotein genes. PMID- 11493598 TI - An essential function of the mitochondrial sulfhydryl oxidase Erv1p/ALR in the maturation of cytosolic Fe/S proteins. AB - Biogenesis of Fe/S clusters involves a number of essential mitochondrial proteins. Here, we identify the essential Erv1p of Saccharomyces cerevisia mitochondria as a novel component that is specifically required for the maturation of Fe/S proteins in the cytosol, but not in mitochondria. Furthermore, Erv1p was found to be important for cellular iron homeostasis. The homologous mammalian protein ALR ('augmenter of liver regeneration'), also termed hepatopoietin, can functionally replace defects in Erv1p and thus represents the mammalian orthologue of yeast Erv1p. Previously, a fragment of ALR was reported to exhibit an activity as an extracellular hepatotrophic growth factor. Both Erv1p and full-length ALR are located in the mitochondrial intermembrane space and represent the first components of this compartment with a role in the biogenesis of cytosolic Fe/S proteins. It is likely that Erv1p/ALR operates downstream of the mitochondrial ABC transporter Atm1p/ABC7/Sta1, which also executes a specific task in this essential biochemical process. PMID- 11493599 TI - Gastrulation defective, a complement factor C2/B-like protease, interprets a ventral prepattern in Drosophila. AB - gastrulation defective (gd) encodes a serine protease required for specification of dorsal-ventral cell fates during Drosophila embryogenesis. Using RNA microinjection, I show that wild-type gd RNA can restore ventrolateral pattern elements with correct polarity with respect to egg shape in embryos lacking gd function. While low RNA concentrations restore ventrolateral pattern elements, higher concentrations ventralize the embryo. Gastrulation defective concentration has a rate-limiting effect on the domain of high Dorsal concentration but little effect upon the slope of the gradient. In embryos from pipe-null females, much higher RNA concentrations generate an ectopic axis oriented with respect to the site of injection. The data suggest that the Dorsal gradient is not directly determined by asymmetric cues in the eggshell but arises de novo within the perivitelline space as a consequence of self-regulatory properties of the protease cascade. A homology to the mammalian complement factors C2 and B is also described. PMID- 11493600 TI - A kinase-independent function of Ask1 in caspase-independent cell death. AB - Ask1 (apoptosis signal-regulating kinase 1) is activated as a consequence of cell exposure to a variety of stresses and can then initiate apoptosis. A known pathway of apoptosis downstream of Ask1 involves the activation of the stress activated protein kinases, the release of cytochrome c from mitochondria, the activation of caspases, and the fragmentation of nuclei. Here, we characterized a novel mechanism of Ask1-mediated cell killing that is triggered by the interaction with Daxx. Co-transfection of Ask1 and Daxx induced a caspase independent cell-death process characterized at the morphological level by distinctive crumpled nuclei easily distinguishable from the condensed and fragmented nuclei seen during classical caspase-dependent apoptosis. The kinase activity of Ask1 was not involved in this process, because mutants lacking kinase activity were as efficient as wild type Ask1 in mediating Daxx-induced cell death. Ask1N, a deletant that lacks the C-terminal half including the kinase domain of Ask1, was constitutively active in producing crumpled nuclei. In contrast, Ask1DeltaN, the reciprocal deletant that possesses constitutive kinase activity, produced fragmented nuclei typical of caspase-dependent death processes. We conclude that in addition to a caspase-dependent pro-apoptotic function that depends on its kinase activity, Ask1 possesses a caspase independent killing function that is independent on its kinase activity and is activable by interaction with Daxx. In the physiological situation, such an activity is induced as a consequence of the translocation of Daxx from the nucleus to the cytoplasm, a condition that occurs following activation of the death receptor Fas. PMID- 11493601 TI - The iota-carrageenase of Alteromonas fortis. A beta-helix fold-containing enzyme for the degradation of a highly polyanionic polysaccharide. AB - Carrageenans are gel-forming hydrocolloids extracted from the cell walls of marine red algae. They consist of d-galactose residues bound by alternate alpha(1 ->3) and beta(1-->4) linkages and substituted by one (kappa-carrageenan), two (iota-carrageenan), or three (lambda-carrageenan) sulfate-ester groups per disaccharide repeating unit. Both the kappa- and iota-carrageenan chains adopt ordered conformations leading to the formation of highly ordered aggregates of double-stranded helices. Several kappa-carrageenases and iota-carrageenases have been cloned from marine bacteria. Kappa-carrageenases belong to family 16 of the glycoside hydrolases, which essentially encompasses polysaccharidases specialized in the hydrolysis of the neutral polysaccharides such as agarose, laminarin, lichenan, and xyloglucan. In contrast, iota-carrageenases constitute a novel glycoside hydrolase structural family. We report here the crystal structure of Alteromonas fortis iota-carrageenase at 1.6 A resolution. The enzyme folds into a right-handed parallel beta-helix of 10 complete turns with two additional C terminal domains. Glu(245), Asp(247), or Glu(310), in the cleft of the enzyme, are proposed as candidate catalytic residues. The protein contains one sodium and one chloride binding site and three calcium binding sites shown to be involved in stabilizing the enzyme structure. PMID- 11493602 TI - Amides are novel protein modifications formed by physiological sugars. AB - The Maillard reaction, or nonenzymatic browning, proceeds in vivo, and the resulting protein modifications (advanced glycation end products) have been associated with various pathologies. Despite intensive research only very few structures have been established in vivo. We report here for the first time N(6) [2-[(5-amino-5-carboxypentyl)amino]-2-oxoethyl]lysine (GOLA) and N(6) glycoloyllysine (GALA) as prototypes for novel amide protein modifications produced by reducing sugars. Their identity was confirmed by independent synthesis and coupled liquid chromatography/mass spectrometry. Model reactions with N(alpha)-t-butoxycarbonyl-lysine showed that glyoxal and glycolaldehyde are immediate precursors, and reaction pathways are directly linked to N(epsilon) carboxymethyllysine via glyoxal-imine structures. GOLA, the amide cross-link, and 1,3-bis(5-amino-5-carboxypentyl)imidazolium salt (GOLD), the imidazolium cross link, share a common intermediate. The ratio of GOLA to GOLD is greater when glyoxal levels are low at constant lysine concentrations. GOLA and GALA formation from the Amadori product of glucose and lysine depends directly upon oxidation. With the advanced glycation end product inhibitors aminoguanidine and pyridoxamine we were able to dissect oxidative fragmentation of the Amadori product as a second mechanism of GOLA formation exactly coinciding with N(epsilon)-carboxymethyllysine synthesis. In contrast, the formation of GALA appears to depend solely upon glyoxal-imines. After enzymatic hydrolysis GOLA was found at 66 pmol/mg of brunescent lens protein. This suggests amide protein modifications as important markers of pathophysiological processes. PMID- 11493603 TI - Ovocalyxin-32, a novel chicken eggshell matrix protein. isolation, amino acid sequencing, cloning, and immunocytochemical localization. AB - The eggshell is a highly ordered structure resulting from the deposition of calcium carbonate concomitantly with an organic matrix upon the eggshell membranes. Mineralization takes place in an acellular uterine fluid, which contains the ionic and matrix precursors of the eggshell. We have identified a novel 32-kDa protein, ovocalyxin-32, which is expressed at high levels in the uterine and isthmus regions of the oviduct, and concentrated in the eggshell. Sequencing of peptides derived from the purified protein allowed expressed sequence tag sequences to be identified that were assembled to yield a full length composite sequence whose conceptual translation product contained the complete amino acid sequence of ovocalyxin-32. Data base searches revealed that ovocalyxin-32 has limited identity (32%) to two unrelated proteins: latexin, a carboxypeptidase inhibitor expressed in the rat cerebral cortex and mast cells, and a skin protein, which is encoded by a retinoic acid receptor-responsive gene, TIG1. High level expression of ovocalyxin-32 was limited to the isthmus and uterus tissue, where immunocytochemistry at the light and electron microscope levels demonstrated that ovocalyxin-32 is secreted by surface epithelial cells. In the eggshell, ovocalyxin-32 localizes to the outer palisade layer, the vertical crystal layer, and the cuticle of the eggshell, in agreement with its demonstration by Western blotting at high levels in the uterine fluid during the termination phase of eggshell formation. Ovocalyxin-32 is therefore identified as a novel protein synthesized in the distal oviduct where hen eggshell formation occurs. PMID- 11493604 TI - The coatomer-interacting protein Dsl1p is required for Golgi-to-endoplasmic reticulum retrieval in yeast. AB - Sec22p is an endoplasmic reticulum (ER)-Golgi v-SNARE protein whose retrieval from the Golgi compartment to the endoplasmic reticulum (ER) is mediated by COPI vesicles. Whether Sec22p exhibits its primary role at the ER or the Golgi apparatus is still a matter of debate. To determine the role of Sec22p in intracellular transport more precisely, we performed a synthetic lethality screen. We isolated mutant yeast strains in which SEC22 gene function, which in a wild type strain background is non-essential for cell viability, has become essential. In this way a novel temperature-sensitive mutant allele, dsl1-22, of the essential gene DSL1 was obtained. The dsl1-22 mutation causes severe defects in Golgi-to-ER retrieval of ER-resident SNARE proteins and integral membrane proteins harboring a C-terminal KKXX retrieval motif, as well as of the soluble ER protein BiP/Kar2p, which utilizes the HDEL receptor, Erd2p, for its recycling to the ER. DSL1 interacts genetically with mutations that affect components of the Golgi-to-ER recycling machinery, namely sec20-1, tip20-5, and COPI-encoding genes. Furthermore, we demonstrate that Dsl1p is a peripheral membrane protein, which in vitro specifically binds to coatomer, the major component of the protein coat of COPI vesicles. PMID- 11493605 TI - Structural and mutational analysis of the PhoQ histidine kinase catalytic domain. Insight into the reaction mechanism. AB - PhoQ is a transmembrane histidine kinase belonging to the family of two-component signal transducing systems common in prokaryotes and lower eukaryotes. In response to changes in environmental Mg(2+) concentration, PhoQ regulates the level of phosphorylated PhoP, its cognate transcriptional response-regulator. The PhoQ cytoplasmic region comprises two independently folding domains: the histidine-containing phosphotransfer domain and the ATP-binding kinase domain. We have determined the structure of the kinase domain of Escherichia coli PhoQ complexed with the non-hydrolyzable ATP analog adenosine 5'-(beta,gamma imino)triphosphate and Mg(2+). Nucleotide binding appears to be accompanied by conformational changes in the loop that surrounds the ATP analog (ATP-lid) and has implications for interactions with the substrate phosphotransfer domain. The high resolution (1.6 A) structure reveals a detailed view of the nucleotide binding site, allowing us to identify potential catalytic residues. Mutagenic analyses of these residues provide new insights into the catalytic mechanism of histidine phosphorylation in the histidine kinase family. Comparison with the active site of the related GHL ATPase family reveals differences that are proposed to account for the distinct functions of these proteins. PMID- 11493606 TI - N-end rule specificity within the ubiquitin/proteasome pathway is not an affinity effect. AB - The N-end rule relates the amino terminus to the rate of degradation through the ubiquitin/26 S proteasome pathway. Proteins bearing basic (type 1) or large hydrophobic (type 2) amino termini are assumed to be targeted through this pathway by their higher affinity for binding to the responsible E3 ligase compared with proteins bearing other residues (type 3). Paradoxically, a significant fraction of eukaryotic protein degradation occurs through the N-end rule pathway, although the majority of cellular proteins are type 3 substrates. We have exploited specific interactions between ubiquitin carrier proteins (E2/Ubc) and their cognate E3 ligases to purify for the first time the mammalian N-end rule ligase E3alpha/Ubr1 to near homogeneity. In vitro studies show that E3alpha forms lysine 48-linked polyubiquitin degradation signals on type 1-3 substrates and is absolutely dependent on Ubc2/Rad6 orthologs. Biochemically defined kinetic studies show that the basis of N-end rule specificity is a k(cat) rather than the K(m) effect originally proposed, since all three substrate classes show similar binding affinities (K(m) approximately 5 microm) but V(max) values that are 100- and 50-fold greater for type 1 and 2 versus type 3 model substrates, respectively. In addition, the N-end rule dipeptides lysylalanine and phenylalanylalanine are general noncompetitive inhibitors for E3alpha-catalyzed ubiquitination of type 1-3 substrates rather than type-specific competitive inhibitors as predicted. These observations are consistent with a model in which the N-end rule effect reflects substrate binding-induced transitions in E3alpha to a catalytically competent conformer, the equilibrium for which depends on the identity of the amino terminus or the presence of basic or hydrophobic surface features. The model reconciles conflicts between specific predictions and empirical observations relating N-end rule targeting in addition to explicating the efficacy of selected dipeptides as potent in vivo inhibitors of this pathway. PMID- 11493607 TI - Functional heterogeneity in liver and lung macrophages. AB - Although initially considered merely "scavenger cells" that participate in immunologic responses only after B and T lymphocytes have performed their biological tasks, more recent evidence suggests that macrophages play a key role in host defense as well as in the maintenance of normal tissue structure and function. For macrophages to perform their biological functions, they must be activated. This involves up-regulation of an array of signaling pathways resulting in altered gene expression and increased biochemical and functional activity. Macrophages have been identified in almost all tissues of the body. However, the basal activity of these cells, as well as their ability to respond to inflammatory mediators, varies considerably with their location. In addition, even within a particular tissue, there is evidence of macrophage heterogeneity. The largest populations of macrophages in the body are located in the liver and lung. Because of the unique attributes of these tissues, hepatic and pulmonary macrophages play essential roles not only in nonspecific host defense but also in the homeostatic responses of these tissues. In this review, the functional and biochemical activities of macrophages localized in the liver and lungs are compared. Evidence suggests that these represent distinct cell populations with unique functions and responsiveness to inflammatory agents. PMID- 11493608 TI - Leukocyte recruitment at sites of tumor: dissonant orchestration. AB - Biopsies of tumors responding to interleukin 2 (IL-2) based immunotherapy have been reported to show a leukocytic infiltration. Clinical responses to IL-2-based immunotherapy, however, are limited, suggesting a failure of leukocyte localization at tumor sites. Leukocyte infiltration at inflammatory sites requires local activation of leukocytes and endothelial cells in a coordinated and defined temporal sequence. There is evidence supporting the theory that infiltration of leukocytes at tumor sites is suboptimal due to a failure of coordination of these localizing events. In this review, factors involved in leukocyte recruitment at sites of inflammation and the coordination of these factors in a successful model of inflammation, i.e., wound healing, are discussed. This example is contrasted with events at tumor sites where alterations in expression of cell adhesion molecules or in the production of activating agents may be present. Additionally, the systemic administration of an activating cytokine such as IL-2 may fail to duplicate events that normally occur within a local environment. These observations may facilitate the design of future immunotherapy trials. PMID- 11493609 TI - Insights into molecular mechanisms of contact hypersensitivity gained from gene knockout studies. AB - Contact hypersensitivity (CHS), a dendritic-cell (DC)-dependent, T-cell-mediated skin immune response to reactive haptens, has been a subject of intense research for many years. The molecular mechanisms underlying CHS are complicated and are not fully understood. During the past few years, varieties of gene-targeted knockout mice have been used in the study of CHS. Such studies have contributed significantly to our understanding of the mechanisms responsible for the initiation of CHS. This review focuses on insights into molecular requirements for CHS gained from knockout studies. PMID- 11493610 TI - IL-1 and TNF independent pathways mediate ICAM-1/VCAM-1 up-regulation in ischemia reperfusion injury. AB - In vitro studies have suggested that targeting interleukin (IL)-1 and tumor necrosis factor (TNF) can be used to regulate intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) and potentially treat kidney inflammation. We therefore evaluated ICAM-1 and VCAM-1 regulation in knockout (KO) mice deficient in both IL-1 receptor 1 (R1) and TNF-R1 during renal ischemia reperfusion injury. ICAM-1 and VCAM-1 mRNA expression was measured with specific murine probes and Northern blotting (n =4/group). Protein expression was measured using immunohistochemistry. Serum creatinine (SCr), tubular histology, and neutrophil infiltration into postischemic kidneys were also quantified. ICAM 1 and VCAM-1 mRNA expression increased in both wild-type (WT) and KO mice at 2, 6, and 24 h. Protein expression of ICAM-1 and VCAM-1 was also increased at 24 h postischemia. SCr levels and tubular necrosis scores were comparable in WT and KO mice at 24 and 48 h. Neutrophil migration in KO mice was decreased at 24 h but comparable to WT at 48 h. These data demonstrate that IL-1 and TNF are not essential for postischemic increases in ICAM-1 and VCAM-1. PMID- 11493611 TI - Reactive oxygen species mediate angiotensin II-induced leukocyte-endothelial cell interactions in vivo. AB - Chronically elevated angiotensin II (Ang-II)-induced hypertension is partly mediated by superoxide production. In this study, we have investigated whether the leukocyte-endothelial cell interactions elicited by Ang-II involve reactive oxygen species (ROS) generation. Intravital microscopy within the rat mesenteric microvessels was used. Superfusion (60 min) with Ang-II (1 nM) induced significant increases in leukocyte rolling flux, adhesion, and emigration, which were inhibited by pretreatment with superoxide dismutase or catalase. Dihydrorhodamine-123 oxidation indicated that ROS are primarily produced by the vessel wall. Administration of dimethylthiourea, desferrioxamine, or N acetylcysteine provoked significant reductions in Ang-II-induced leukocyte endothelial cell interactions. In addition, a blockade of platelet-activating factor or leukotrienes also attenuated such responses significantly. The results presented indicate that in vivo Ang-II-induced leukocyte recruitment is dependent on the generation of intra- and extracellular ROS. Therefore, the use of anti oxidants might constitute an alternative therapy for the control of the subendothelial leukocyte infiltration associated with hypertension and atherosclerosis. PMID- 11493612 TI - IgE-regulated loss, not IgE-regulated synthesis, controls expression of FcepsilonRI in human basophils. AB - Expression of the high-affinity receptor on basophils and mast cells is modulated by immunoglobulin E (IgE) antibody. Recent studies have shown that modulation occurs through interaction of IgE with the receptor itself, but the mechanisms underlying this control are not understood. Taking both a theoretical and experimental approach, we examined several competing models that focus on whether there is IgE-regulated loss, IgE-regulated synthesis, or both regulated loss and synthesis of the Fc receptor for IgE (FcepsilonRI). We report that removing IgE from occupied FcepsilonRI resulted in an accelerated loss only in the unoccupied receptor, with no loss of occupied receptors and no loss of total receptors when all receptors were occupied. Together with previous studies, these results establish that there was IgE-regulated loss of receptors. An examination of synthetic rates of FcepsilonRIalpha using pulse-labeling with (35)S-methionine indicated no difference in synthetic rates in the presence or absence of IgE. Similarly, the presence or absence of IgE had no influence on the levels of mRNA for either alpha, beta, or gamma subunits of FcepsilonRI. Using model simulations, we found that regulated-synthesis models could be distinguished from regulated-loss/constant-synthesis models on the basis of the relationship between starting FcepsilonRI densities and changes in density after culture for 1 week in the absence of IgE. Experimental data from this type of study fit a regulated loss model that did not include regulation of synthesis. Taken together, these results suggest that IgE regulates cell surface expression of FcepsilonRI only by regulating the rate that receptor is lost from the cell surface. PMID- 11493613 TI - Expression of CD94 and NKG2 molecules on human CD4(+) T cells in response to CD3 mediated stimulation. AB - We investigated the ability of human peripheral CD4(+) cells to express CD94 and NKG2 molecules as a consequence of CD3-mediated activation. Using highly purified peripheral CD4(+) T cells, we found expression of both CD94 and NKG2A 15 days after CD3-mediated stimulation of cells. We also determined by reverse transcriptase-PCR that all gene members of NKG2 family-namely, NKG2A, -C, -D, and -E-are sequentially expressed on CD4(+) cells. We found that this expression is tightly regulated by cytokines, and we identified transforming growth factor beta1 and interleukin-10 as the main factors that, on CD3-dependent stimulation, positively contribute to the expression of CD94 and NKG2A on CD4(+) cells. We also investigated the functional role of NKG2A and found that coligation of CD3 and NKG2A by specific monoclonal antibodies results in significant inhibition of interferon gamma and tumor necrosis factor alpha production by stimulated CD4(+) cells. The presence and function of these receptors on CD4(+) lymphocytes support a more general role for NKG2 molecules, whose functions were originally thought to be confined to cytotoxic cells, in the immune system. PMID- 11493614 TI - Growth factor regulation of neutrophil-endothelial cell interactions. AB - The effects of the angiogenic factors basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) on human polymorphonuclear leukocyte (PMNL)-endothelial cell adhesion and transendothelial migration (TEM) were investigated. Stimulation of human umbilical vein endothelial cells by VEGF or bFGF for 18 h up-regulated intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 expression and significantly increased PMNL adhesion and TEM in response to complement fragment 5a (C5a) or interleukin (IL)-8. In contrast, continued exposure to bFGF (24 h-6 days) down-regulated basal and IL-1- or tumor necrosis factor (TNF)-induced intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and E-selectin expression as well as PMNL adhesion and TEM. These effects could be reversed by introduction of high concentrations of TNF alpha, C5a, or IL-8. None of these inhibitory effects was observed with VEGF. The acute effects of bFGF and VEGF may facilitate PMNL emigration during acute inflammation, but continued bFGF production may have anti-inflammatory actions during chronic inflammation, angiogenesis, and tumor defense by inhibition of endothelial activation for leukocyte recruitment. PMID- 11493615 TI - Voltage-dependent and calcium-activated ion channels in the human mast cell line HMC-1. AB - The mechanisms underlying the recruitment, differentiation, and sustained activation of mast cells in disease are likely to include modulation of ion channels. Specific Ca(2+), K(+), and Cl(-) conductances have been identified in rodent mast cells, but there are no equivalent data on human mast cells. We have used the whole-cell patch-clamp technique to characterize macroscopic ion currents in both the human mast cell line HMC-1 and human skin mast cells (HSMCs) at rest and in HMC-1 after activation with calcium ionophore. HSMCs were electrically silent at rest. In contrast, HMC-1 expressed a strong outwardly rectifying voltage-dependent Cl(-) conductance characteristic of ClC-4 or ClC-5 and a small inwardly rectifying K(+) current not carried by the classical Kir family of K(+) channels. Calcium ionophore induced the appearance of outwardly rectifying Ca(2+)-activated Cl(-) and K(+) currents, while hypotonicity induced another outwardly rectifying conductance typical of ClC-3. Reverse transcription PCRs confirmed that mRNAs for the voltage-dependent Cl(-) channels ClC-3 and -5 were expressed. This is the first definitive description of a ClC-4/5-like current in a native leukocyte. We suggest that this current may contribute to the malignant phenotype while the Ca(2+)-activated K(+) and Cl(-) currents may be involved in cell activation. PMID- 11493616 TI - Monocyte differentiation in intestine-like macrophage phenotype induced by epithelial cells. AB - Macrophages in normal colonic mucosa show a specific and distinct phenotype with low expression of the typical monocyte/macrophage surface antigens CD14, CD16, and CD11b and T-cell costimulatory molecules. A method for the in vitro induction of a macrophage phenotype similar to this intestinal phenotype is presented. Multicellular spheroids (MCSs) of intestinal epithelial cell (IEC) and control cell lines were cocultured with elutriated monocytes. Surface antigen expression was analyzed by immunohistochemistry and flow cytometry. Interleukin (IL)-1beta mRNA was measured by quantitative PCR. Monocytes adhered and infiltrated the MCSs within 24 h. In the MCSs of all IEC lines, the typical monocyte/macrophage surface antigens CD14, CD16, CD11b, and CD11c, which are detectable after 24 h of coculture by immunohistochemistry and flow cytometry, were down-regulated after 7 days (e.g., for CD14 at 24 h, expression was 86% of CD33+ cells; at day 7, it was 11%). A clear decrease of lipopolysaccharide (LPS)-stimulated IL-1beta transcription in monocytes cocultured with IEC MCSs could be observed during the 7-day period. For the first time an intestine-like macrophage-phenotype could be induced in vitro. Interactions with IECs play an essential role during this differentiation, which is of functional relevance, e.g., for LPS-induced cytokine secretion. PMID- 11493617 TI - Feedback activation of T-cell antigen-presenting cells during interactions with T cell responders. AB - Like many T cells in the myelin basic protein (MBP)-specific T-cell repertoire, CD4(-) GP2.3H3.16 (3H3) T cells recognize guinea pig MBP as an agonist but recognize autologous rat (R)MBP as a mixed agonist/antagonist. 3H3 T cells do not exhibit proliferative responses to RMBP but nonetheless respond to RMBP by accumulation of T-cell surface I-A/peptide complexes and generation of T-cell antigen-presenting cell (T-APC) activity. This study showed that presentation of RMBP by 3H3 T-APC is long-lived but is lost during interactions with cognate responders or on overt activation of T-APCs. Presentation of RMBP to encephalitogenic T cells resulted in the reciprocal activation of 3H3 T-APCs as evidenced by blastogenesis, proliferation, and induction of interleukin-2R and OX40 markers on 3H3 T-APC. These data indicate that T-APCs, like B-cell APCs, undergo clonal expansion after presentation of a cognate antigen to T-cell responders. PMID- 11493618 TI - Effects of hypertonic saline on expression of human polymorphonuclear leukocyte adhesion molecules. AB - Hypertonic saline prevents vascular adherence of neutrophils and ameliorates ischemic tissue injury. We hypothesized that hypertonic saline attenuates N formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated expression of adhesion molecules on human polymorphonuclear leukocytes (PMNLs). fMLP-stimulated up regulation of beta2-integrins was diminished by hypertonic saline but not by hypertonic choline chloride-, mannitol-, or sucrose-modified Hanks' buffered salt solution. Shedding of L-selectin was decreased by hypertonic saline and choline chloride but not by hypertonic mannitol or sucrose. When the effects of hypertonic sodium chloride- and choline chloride-modified media were compared, neither solution affected fMLP-receptor binding but both equally inhibited fMLP stimulated increase in intracellular calcium, ionophore A23187, and phorbol myristate acetate (PMA)-stimulated numerical up-regulation of beta2-integrins. Analysis of mitogen-activated protein (MAP) kinases p38 and p44/42 for phosphorylation revealed that hypertonic solutions did not differ in preventing fMLP-stimulated increases in phospho-p38 and phospho-p44/42. Resting PMNLs shrunk by hypertonic saline increased their volume during incubation and further during chemotactic stimulation. Addition of amiloride further enhanced inhibition of up regulation of beta2-integrins. No fMLP-stimulated volume changes occurred in PMNLs exposed to hypertonic choline chloride, resulting in significant cell shrinkage. Results suggest a sodium-specific inhibitory effect on up-regulation of beta2-integrins of fMLP-stimulated PMNLs, which is unlikely to be caused by alterations of fMLP receptor binding, decrease in cytosolic calcium, attenuation of calcium or protein kinase C-dependent pathways, suppression of p38- or p44/42 MAP kinase-dependent pathways, or cellular ability to increase or decrease volumes. PMID- 11493619 TI - Deleterious effects of Echinacea purpurea and melatonin on myeloid cells in mouse spleen and bone marrow. AB - The neurohormone, melatonin, a product of the pineal gland, is a potent immune cell stimulant. Phytochemicals contained in root extracts of the plant species Echinacea purpurea are also potent as immune cell stimulants. Both agents are potent stimulants of T, B, and/or natural killer cells, but little is known of their effect on other hemopoietic cells, specifically granular leukocytes, also participants in a wide variety of disease defense processes. Given their current popularity and availability for amelioration of a) jet lag and sleep disorders (melatonin) and b) virus-mediated respiratory infections (E. purpurea), we investigated the effects of these agents on granular leukocytes and their precursors, myeloid cells. Mice received these agents daily for 7 or 14 days via the diet, thus mimicking human administration, after which spleens and bone marrow were removed and assessed for mature, differentiated granulocytes and their myeloid progenitors. The influence of these agents was directly related to the stage of cell maturity. Administration of both agents together resulted in significantly elevated levels of myeloid progenitor cells in both bone marrow and spleen and significantly reduced levels of mature, functional granulocyte progeny in both organs, suggesting a) increased precursor proliferation, b) antiapoptosis among the progenitors, and/or c) inhibition of precursor maturation-the latter readily explaining the paucity of mature granulocyte progeny. In conclusion, individual administration of either the herbal derivative and melatonin was either without effect (E. purpurea) or even advantageous (melatonin) to cells of this lineage, but when administered together, these agents significantly perturbed myelopoiesis. PMID- 11493620 TI - MC148 encoded by human molluscum contagiosum poxvirus is an antagonist for human but not murine CCR8. AB - The viral CC chemokines MC148, encoded by the poxvirus molluscum contagiosum, and viral macrophage inflammatory protein (vMIP)-I and vMIP-II, encoded by human herpesvirus 8, were probed on the murine CC receptor (CCR) 8 in parallel with human CCR8. In calcium mobilization assays, vMIP-I acted as a high-affinity agonist, whereas vMIP-II acted as a low-affinity antagonist on the murine CCR8 as well as the human CCR8. MC148 was found to bind and block responses through the human CCR8 with high affinity, but surprisingly MC148 was unable to bind and block responses through the murine CCR8. Because MC148 is the only high-affinity antagonist known to target and be selective for CCR8, MC148 is a valuable tool to decipher the role played by CCR8 in the immune system. This study shows that MC148 could not be used in murine inflammatory models; however, it will be interesting to see whether it can be used in other animal models to delineate the role played by CCR8. PMID- 11493621 TI - Contributions of Neisseria meningitidis LPS and non-LPS to proinflammatory cytokine response. AB - To determine the relative contribution of lipopolysaccharide (LPS) and non-LPS components of Neisseria meningitidis to the pathogenesis of meningococcal sepsis, this study quantitatively compared cytokine induction by isolated LPS, wild-type serogroup B meningococci (strain H44/76), and LPS-deficient mutant meningococci (strain H44/76[pLAK33]). Stimulation of human peripheral-blood mononuclear cells with wild-type and LPS-deficient meningococci showed that non-LPS components of meningococci are responsible for a substantial part of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta production and virtually all interferon (IFN)-gamma production. Based on tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of LPS in proteinase K-treated lysates of N. meningitidis H44/76, a quantitative comparison was made between the cytokine inducing capacity of isolated and purified LPS and LPS-containing meningococci. At concentrations of >10(7) bacteria/mL, intact bacteria were more potent cytokine inductors than equivalent amounts of isolated LPS, and cytokine induction by non-LPS components was additive to that by LPS. Experiments with mice showed that non-LPS components of meningococci were able to induce cytokine production and mortality. The principal conclusion is that non-LPS parts of N. meningitidis may play a role in the pathogenesis of meningococcal sepsis by inducing substantial TNF-alpha, IL-1beta, and IFN-gamma production. PMID- 11493622 TI - Augmented TNF-alpha and IL-10 production by primed human monocytes following interaction with oxidatively modified autologous erythrocytes. AB - The presence of dysfunctional/damaged red blood cells (RBCs) has been associated with adverse clinical effects during the inflammatory response. The aim of this study was to elucidate whether oxidatively modified, autologous RBCs modulate monocyte cytokine responses in humans. Monocyte tumor necrosis factor alpha (TNF alpha) and IL-10 production was measured in whole blood from healthy volunteers using ELISA and flow cytometry. Oxidatively modified RBCs (15 mM phenylhydrazine, 1 h, OX-RBC) or vehicle-treated RBCs (VT-RBC) opsonized by autologous serum were administered alone or in combination with one of three priming agents: E. coli lipopolysaccharide (LPS, 0.2 ng/ml), zymosan A (1 mg/ml), or phorbol 12-myristate 13-acetate (PMA, 50 ng/ml). OX-RBC or VT-RBC alone did not result in the release of TNF-alpha or IL-10. LPS, zymosan, and PMA caused marked and dose-dependent increases in TNF-alpha and IL-10 production. Addition of OX-RBC augmented the LPS , zymosan-, and PMA-induced TNF-alpha release by approximately 100%. OX-RBC augmented LPS- and zymosan-induced IL-10 release by 400-600%. Flow cytometry analyses showed that monocytes were responsible for TNF-alpha and IL-10 production in whole blood. The presence of OX-RBC alone increased the complexity of CD14+ monocytes but caused no cytokine production. LPS alone induced cytokine production without altering cell complexity. After the combined (OX-RBC+LPS) treatment, monocytes of high complexity were responsible for TNF-alpha production. The presence of mannose or galactose (at 10-50 mM) did not alter the observed augmentation of cytokine production by OX-RBC, suggesting that lectin receptors are not involved in the response. These studies indicate that the interaction between damaged autologous erythrocytes and monocytes has a major impact on the cytokine responses in humans. An augmented cytokine production by the mononuclear phagocyte system may adversely affect the clinical course of injury and infections especially in genetic or acquired RBC diseases or after transfusions. PMID- 11493623 TI - Early effects of insulin-like growth factor-1 in activated human T lymphocytes. AB - This study evaluates the effects of insulin-like growth factor (IGF)-1 receptor (IGF-1R) down-regulation in stimulated T lymphocytes by investigating the expression of early activation proteins CD69, CD25, and interleukin (IL)-2. We found that IGF-1 does not modify CD69 expression but increases transcription and protein synthesis of CD25 and IL-2. The lowest level of IGF-1R detected after 15 min of activation suggested that the effects of IGF-1 occur at the initiation of cell activation. The activation of IGF-1R was confirmed by IGF-1R phosphorylation and increased phosphorylation of microtubule-associated protein kinase. We also detected the alternative IGF-1 transcripts Ea, with paracrine/autocrine regulation, and Eb, with endocrine regulation, in Jurkat cells and in quiescent T lymphocytes, and we detected IGF-1 protein in the culture medium after stimulation. These data suggest that the proliferative effects of IGF-1 on T lymphocytes include both autocrine/paracrine and endocrine processes. PMID- 11493624 TI - A novel bioactive 31-amino acid endothelin-1 is a potent chemotactic peptide for human neutrophils and monocytes. AB - Endothelin (ET)-1(1-31) is a novel 31-amino acid-length peptide derived from big ET-1 by chymase or other chymotrypsin-type proteases and is a major ET derivative in human neutrophils. In this study, we revealed that ET-1(1-31), but not big ET, exhibited chemotactic activities toward human neutrophils and monocytes as an inflammatory mediator, although the effects were less potent than those of formyl methionyl-leucyl-phenylalanine or interleukin-8. However, the chemotactic effects of ET-1(1-31) were much greater than those of the 21-amino acid ET-1, ET-1(1-21). Checkerboard analyses revealed that the effects are chemotactic rather than chemokinetic. The effects of ET-1(1-31) are not mediated by interleukin-8 or monocyte chemoattractant protein-1. The chemotactic effects and an increase in intracellular-free Ca(2)(+) caused by ET-1(1-31) were significantly inhibited by BQ123, an ET(A) receptor antagonist, but not by BQ788, an ET(B) receptor antagonist, suggesting that ET-1(1-31) mediates chemotaxis through an ET(A) or ET(A)-like receptor. PMID- 11493625 TI - Functional HIV CXCR4 coreceptor on human epithelial Langerhans cells and infection by HIV strain X4. AB - HIV can cross the intact epithelium of genital mucosae via Langerhans cells. Fresh Langerhans cells are known to express CD4 and CCR5. The presence of CXCR4 on the surface of cultured but not freshly isolated Langerhans cells has been described. In the present study, we demonstrate that CXCR4 was expressed by fresh Langerhans cells isolated and purified from epidermis. However, the percentage of Langerhans cells expressing CXCR4 or CCR5 increased during maturation of the cells in culture, especially in the presence of exogenous granulocyte-macrophage colony-stimulating factor. To determine whether CXCR4 was functional, freshly isolated Langerhans cells were infected with HIV LAI, a T-cell-tropic strain, and p24 protein production was measured in culture supernatants. p24 production was observed when infected Langerhans cells were cocultured with SupT1 cells. However, the presence of HIV provirus DNA was evidenced within the infected Langerhans cells by nested PCR. Ultrastructural studies confirmed the formation of syncytia when Langerhans cells were cocultured with SupT1 cells. Preincubation of Langerhans cells with azidothymidine or SDF-1-alpha, a natural ligand for CXCR4, prevented infection. These data demonstrated that CXCR4 is present on the surface of Langerhans cells freshly isolated from human skin epidermis and that this expression is functional. PMID- 11493626 TI - FcgammaR-mediated phagocytosis by human macrophages involves Hck, Syk, and Pyk2 and is augmented by GM-CSF. AB - The receptors for the constant region of immunoglobulin G (FcgammaR) are widely expressed on cells of hemopoietic lineage and plays an important role in host defense. We investigated the signaling pathways during FcgammaR-mediated phagocytosis in human monocyte-derived macrophages (MDMs) and examined the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on these events. FcgammaR-mediated phagocytosis resulted in enhanced tyrosine phosphorylation of a wide range of cellular proteins and activation of tyrosine kinases Hck, Syk, and Pyk2, as well as the multidomain adapter protein paxillin. Stimulation of MDMs with GM-CSF augmented FcgammaR-mediated phagocytosis and increased the levels of tyrosine phosphorylation in phagocytosing MDM cultures, indicating tyrosine kinase-mediated activation. GM-CSF treatment of MDMs without a phagocytic stimulus did not activate Syk, suggesting that GM-CSF may act either distally to Syk in the FcgammaR-mediated signaling cascade or on a parallel pathway activated by the FcgammaR. This study shows that early signaling events during FcgammaR mediated phagocytosis in human MDMs involve activation of Syk, Hck, and paxillin. It also provides the first evidence for Pyk2 activation during phagocytosis and a baseline for further studies on the effect of GM-CSF on FcgammaR-mediated phagocytosis. PMID- 11493627 TI - A small-molecule antagonist of LFA-1 blocks a conformational change important for LFA-1 function. AB - Lymphocyte function-associated antigen (LFA)-1/intercellular adhesion molecule (ICAM)-1 interactions mediate several important steps in the evolution of an immune response. LFA-1 is normally expressed in a quiescent state on the surface of leukocytes and interacts weakly with its ligands ICAM-1, -2, and -3. LFA-1 activity may be regulated by receptor clustering and by increasing the affinity of LFA-1 for its ligands. Affinity modulation of LFA-1 has been shown to occur via a conformational change in the LFA-1 heterodimer that can be detected by using monoclonal antibody 24 (mAb24). We have recently described a small-molecule antagonist of LFA-1, BIRT 377, that demonstrates selective in vitro and in vivo inhibition of LFA-1/ICAM-1-mediated binding events. We now demonstrate that BIRT 377 blocks the induction of the mAb24 reporter epitope on LFA-1 on the surface of SKW-3 cells treated with various agonists known to induce high-affinity LFA-1. These data imply that BIRT 377 exerts its inhibitory effects by preventing up regulation of LFA-1 to its high-affinity conformation. PMID- 11493628 TI - Biliary glycoprotein (BGPa, CD66a, CEACAM1) mediates inhibitory signals. AB - Biliary glycoprotein (BGP, CD66a, CEACAM1) is a member of the carcinoembryonic antigen family (CEA, CD66), a group of transmembrane proteins belonging to the immunoglobulin superfamily. The structural features surrounding the tyrosine residues in the cytoplasmic domain of BGP share similarity with the consensus sequence of the immunoreceptor tyrosine-based inhibition motif (ITIM), the docking site for SHIP, SHP-1, and SHP-2 molecules. Using the well-characterized inhibitory receptor, FcgammaRIIB, we constructed a FcgammaRIIB-BGPa chimeric molecule that contained the extracellular and transmembrane domain of FcgammaRIIB and the cytoplasmic tail of BGPa and expressed it in DT40 B cells. Our results showed that FcgammaRIIB-BGPa, just like the unmodified FcgammaRIIB molecule, inhibited calcium influx in activated DT40 B cells. Substitution of tyrosine with phenylalanine (Y459F) in FcgammaRIIB-BGPa completely abrogated its ability to inhibit calcium influx, indicating that the motif surrounding Y459 is ITIM. The presence of ITIM was also supported by showing that the FcgammaRIIB-BGPa-mediated inhibitory effect was reduced in SHP-1and SHP-2 mutant DT40 B cells and further diminished in a SHP-1/-2 double-deficient mutant line. The results suggest that SHP-1 and SHP-2 are required for the FcgammaRIIB-BGPa-mediated inhibitory signals. PMID- 11493633 TI - Cyclic AMP signalling. PMID- 11493634 TI - The Trio family of guanine-nucleotide-exchange factors: regulators of axon guidance. AB - Axon guidance requires the integration of diverse guidance signals presented by numerous extracellular cues and cell-cell interactions. The molecular mechanisms that interpret these signals involve networks of intracellular signaling proteins that coordinate a variety of responses to the environment, including remodeling and assembly of the actin cytoskeleton. Although it has been clear for some time that Rho family GTPases play a central role in the orchestration of cytoskeletal assembly, our understanding of the components that regulate these important molecules is far more primitive. Recent functional studies of the Trio family of guanine-nucleotide-exchange factors reveal that Trio proteins play a vital role in neuronal cell migration and axon guidance. Although the molecular analysis of Trio proteins is still in its infancy, accumulated evidence suggests that Trio proteins function as integrators of multiple upstream inputs and as activators of multiple downstream pathways. Future studies of these mechanisms promise to yield insights not only into neural development but also into the ongoing function and remodeling of the adult nervous system. PMID- 11493635 TI - Myosin learns to walk. AB - Recent experiments, drawing upon single-molecule, solution kinetic and structural techniques, have clarified our mechanistic understanding of class V myosins. The findings of the past two years can be summarized as follows: (1) Myosin V is a highly efficient processive motor, surpassing even conventional kinesin in the distance that individual molecules can traverse. (2) The kinetic scheme underlying ATP turnover resembles those of myosins I and II but with rate constants tuned to favor strong binding to actin. ADP release precedes dissociation from actin and is rate-limiting in the cycle. (3) Myosin V walks in strides averaging approximately 36 nm, the long pitch pseudo-repeat of the actin helix, each step coupled to a single ATP hydrolysis. Such a unitary displacement, the largest molecular step size measured to date, is required for a processive myosin motor to follow a linear trajectory along a helical actin track. PMID- 11493636 TI - Investigation of the roles of Ca(2+) and InsP(3) diffusion in the coordination of Ca(2+) signals between connected hepatocytes. AB - Glycogenolytic agonists induce coordinated Ca(2+) oscillations in multicellular rat hepatocyte systems as well as in the intact liver. The coordination of intercellular Ca(2+) signals requires functional gap-junction coupling. The mechanisms ensuring this coordination are not precisely known. We investigated possible roles of Ca(2+) or inositol 1,4,5-trisphosphate (InsP(3)) as a coordinating messengers for Ca(2+) spiking among connected hepatocytes. Application of ionomycin or of supra-maximal concentrations of agonists show that Ca(2+) does not significantly diffuse between connected hepatocytes, although gap junctions ensure the passage of small signaling molecules, as demonstrated by FRAP experiments. By contrast, coordination of Ca(2+) spiking among connected hepatocytes can be favored by a rise in the level of InsP(3), via the increase of agonist concentrations, or by a shift in the affinity of InsP(3) receptor for InsP(3). In the same line, coordination cannot be achieved if the InsP(3) is rapidly metabolized by InsP(3)-phosphatase in one cell of the multiplet. These results demonstrate that even if small amounts of Ca(2+) diffuse across gap junctions, they most probably do not play a significant role in inducing a coordinated Ca(2+) signal among connected hepatocytes. By contrast, coordination of Ca(2+) oscillations is fully dependent on the diffusion of InsP(3) between neighboring cells. PMID- 11493637 TI - Life without a cell membrane: regeneration of protoplasts from disintegrated cells of the marine green alga Bryopsis plumosa. AB - When the multi-nucleate giant cells of the green alga Bryopsis plumosa (Huds.) Ag. are injured, the protoplasm is extruded from the cells and can generate spontaneously numerous new cells. The cell organelles aggregate rapidly in seawater and become covered with a gelatinous envelope within 15 minutes. A lipid cell membrane is formed inside the envelope within 9 to 12 hours and about 15% of the original cell membrane is recycled to make the membrane of new protoplasts. Cytochemical studies using Nile Red and various enzymes revealed that the primary envelope is initially composed of polysaccharides, and then transformed into a polysaccharide-lipid complex. Fluorescein diacetate staining showed that the primary envelope has some characteristics of a cell membrane including semi permeability and selective transport of materials. The aggregation of cell organelles appears to be mediated by two kinds of materials, one present in vacuolar sap and the other on the surface of the cell organelles. About a thousand new cells were generated from a single disintegrated branch and 40% of them eventually developed into mature plants. PMID- 11493638 TI - The Drosophila Dpit47 protein is a nuclear Hsp90 co-chaperone that interacts with DNA polymerase alpha. AB - Hsp90 is gaining increasing importance as a protein involved in controlling the normal functioning of the cell. To do this it apparently interacts with a battery of co-chaperone proteins that are involved in both substrate recognition and the progression of the Hsp90 catalytic pathway. In this report we have identified the Drosophila Dpit47 protein (DNA polymerase interacting tpr containing protein of 47 kDa) through its interaction with the DNA polymerase alpha. This protein is a predominantly nuclear protein, which forms a tight and stoichiometric interaction with Hsp90 and shows interaction with Hsp70. It also has substantial homology to other known Hsp90 co-chaperones, e.g. CNS1 and hop1, making it likely that this protein also functions as an Hsp90 co-chaperone. The interaction with the DNA polymerase alpha is not related to the special situation in early embryos where there are large amounts of maternal protein stockpiles of the polymerase, as it occurs to the same level in early and late embryos and also in proliferating cell culture. However, it does not occur in quiescent cells, making it likely that the protein is related to proliferation. This is also consistent with Dpit47 expression being higher in proliferating cells. The interaction between the Dpit47 and the polymerase takes place predominantly in the nucleoplasm, and seems to involve several subunits of the polymerase in comparable amounts, making it unlikely that it is solely required for the assembly of the polymerase complex. The polymerase can also be seen to interact with Hsp90, and the interaction between Dpit47 and the polymerase is increased by the specific Hsp90 inhibitor geldanamycin. This suggests that a complex of the Dpit47, Hsp90 and DNA polymerase exists in the cell. The interaction between DNA polymerase alpha and Dpit47 completely inhibits the activity of the polymerase. These results suggest that Hsp90 acts as a chaperone for DNA polymerase alpha and that this interaction is mediated through the novel co-chaperone Dpit47. This provides the first suggestion of a role for chaperones in DNA replication in higher eukaryotes. PMID- 11493639 TI - DNA replication licensing and human cell proliferation. AB - The convergence point of growth regulatory pathways that control cell proliferation is the initiation of genome replication, the core of which is the assembly of pre-replicative complexes resulting in chromatin being "licensed" for DNA replication in the subsequent S phase. We have analysed regulation of the pre replicative complex proteins ORC, Cdc6, and MCM in cycling and non-proliferating quiescent, differentiated and replicative senescent human cells. Moreover, a human cell-free DNA replication system has been exploited to study the replicative capacity of nuclei and cytosolic extracts prepared from these cells. These studies demonstrate that downregulation of the Cdc6 and MCM constituents of the replication initiation pathway is a common downstream mechanism for loss of proliferative capacity in human cells. Furthermore, analysis of MCM protein expression in self-renewing, stable and permanent human tissues shows that the three classes of tissue have developed very different growth control strategies with respect to replication licensing. Notably, in breast tissue we found striking differences between the proportion of mammary acinar cells that express MCM proteins and those labelled with conventional proliferation markers, raising the intriguing possibility that progenitor cells of some tissues are held in a prolonged G1 phase or "in-cycle arrest". We conclude that biomarkers for replication-licensed cells detect, in addition to actively proliferating cells, cells with growth potential, a concept that has major implications for developmental and cancer biology. PMID- 11493640 TI - Repression of Wnt-5a impairs DDR1 phosphorylation and modifies adhesion and migration of mammary cells. AB - The Wnt-5a gene encodes a secreted protein that controls several normal processes during embryogenesis and development of adult tissues by as yet unknown mechanisms. Endogenous expression of Wnt-5a mRNA is known to occur in both mouse and human mammary cell lines. To investigate the biological role of Wnt-5a in the human mammary epithelial cell line HB2, we used an antisense approach to repress endogenous expression of Wnt-5a protein. We also generated a cell population that constitutively overexpresses this protein. We found that overexpression of Wnt-5a protein enhanced cell-to-collagen binding and abolished hepatocyte growth factor stimulated migration of HB2 transfectants through collagen matrices. Conversely, repression of Wnt-5a protein led to cell scattering, impaired cell-collagen interaction and enhanced cell motility. As we were searching for modified collagen receptors in antisense cells, we discovered that the collagen-binding discoidin domain receptor 1 (DDR1) failed to undergo phosphorylation. In reciprocal experiments, phosphorylation of DDR1 was consistently enabled by expression of Wnt-5a-HA protein in non-Wnt-5a-producing MCF-7 breast cancer cells. Activation of the Wnt/beta-catenin signalling pathway did not influence or mimic the Wnt-5a-mediated effect on DDR1 phosphorylation. These data demonstrate that Wnt-5a protein participates in regulation of adhesion to and migration through collagen and is also a co-factor necessary for collagen-induced activation of DDR1 receptors in mammary epithelial cells. PMID- 11493641 TI - Collagen IV synthesis is restricted to the enteroendocrine pathway during multilineage differentiation of human colorectal epithelial stem cells. AB - The human large intestine is lined by a rapidly renewing epithelial monolayer where cell loss is precisely balanced with cell production. The continuous supply of new cells is produced by undifferentiated multipotent stem cells via a coordinated program of proliferation and differentiation yielding three epithelial lineages: absorptive, goblet and enteroendocrine. Cell-matrix interactions have been suggested to be regulators of the multilineage differentiation program of the colorectal crypt but the expression of matrix proteins or their receptors does not appear to have the subtlety expected for this task. We have developed an in vitro model system of intestinal epithelial stem cells to facilitate the direct analysis of stem cells undergoing lineage commitment and differentiation. Using this culture system, we can now directly investigate the role of cell-matrix signalling in stem-cell decisions. In this study, collagen-IV synthesis has been followed in monolayers of multipotent cells that have been induced to differentiate into absorptive, goblet and enteroendocrine cells. Our experiments demonstrate that commitment to the enteroendocrine lineage is specifically accompanied by the expression of type-IV collagen that remains enteroendocrine-cell associated. Undifferentiated cells, absorptive cells and goblet cells do not express collagen IV. To confirm that the differential lineage-specific expression of collagen IV observed in the model system was representative of the in vivo situation, collagen-IV synthesis was analysed in isolated human colorectal crypts and tissue sections using immunocytochemistry and in situ hybridisation. These studies confirmed the in vitro findings, in that implementation of the enteroendocrine differentiation program involves synthesis and accumulation of a collagen-IV matrix. Thus, human colorectal enteroendocrine cells are unique in the colorectal crypt in that they assemble a cell-associated collagen-IV-rich matrix not observed on other colorectal epithelial cells. This study provides the first evidence for differential matrix synthesis between colorectal epithelial lineages in human colorectal epithelium. The specialised pericellular environment of the enteroendocrine cells might explain some of the unique phenotypic characteristics of this cell lineage. Furthermore, these findings suggest a potential mechanism whereby individual epithelial cells could modulate their cell-matrix signalling even while rapidly migrating in heterogeneous sheets over a shared basement membrane. PMID- 11493642 TI - The interaction of plectin with actin: evidence for cross-linking of actin filaments by dimerization of the actin-binding domain of plectin. AB - Plectin is a major component of the cytoskeleton and is expressed in a wide variety of cell types. It plays an important role in the integrity of the cytoskeleton by cross-linking the three filamentous networks and stabilizing cell matrix and cell-cell contacts. Sequence analysis showed that plectin contains a highly conserved actin-binding domain, consisting of a pair of calponin-like subdomains. Using yeast two-hybrid assays in combination with in vitro binding experiments, we demonstrate that the actin-binding domain of plectin is fully functional and preferentially binds to polymeric actin. The sequences required for actin binding were identified at the C-terminal end of the first calponin homology domain within the actin-binding domain of plectin. We found that the actin-binding domain of plectin is able to bundle actin filaments and we present evidence that this is mediated by the dimerization of this domain. In addition we also show that plectin and another member of the plakin family, dystonin, can heterodimerize by their actin-binding domains. We propose a new mechanism by which plectin and possibly also other actin-binding proteins can regulate the organization of the F-actin network in the cell. PMID- 11493643 TI - Rabbit M cells and dome enterocytes are distinct cell lineages. AB - We have studied the M cell origin and differentiation pathway in rabbit gut associated lymphoid tissues. Micro-dissected domes and epithelium isolated by ethylene diamine tetra acetic acid detachment allowed us to view the whole epithelial surface from the bottom of crypts to the top of domes. We used monoclonal antibodies specific to the apex of either M cells or dome enterocytes, lectins, and antibodies to vimentin in appendix, distal Peyer's patches and caecal patches. The earliest vimentin-labeled M cells were observed in the BrdU positive proliferative zone of dome-associated crypts. Gradual differentiation of the M cell vimentin cytoskeleton started at this site to progressively give rise to the first pocket-forming M cells in the upper dome. Therefore, these mitotic cells of the crypts appear as the direct precursors of M cells. In addition to an early appearance of M cell markers, a regular mosaic-like relative distribution of M cells and dome enterocytes was already detected in the vicinity of crypts, similar to that observed on the lateral surface of domes where functional M cells lie. This constant distribution implies that there is no trans-differentiation of enterocytes to M cells along the crypt-dome axis. Together, these observations provide very strong evidence in favor of an early commitment in crypts of M cell and enterocyte distinct lineages. PMID- 11493644 TI - Sonic hedgehog increases the commitment of pluripotent mesenchymal cells into the osteoblastic lineage and abolishes adipocytic differentiation. AB - The proteins of the hedgehog (Hh) family regulate various aspects of development. Recently, members of this family have been shown to regulate skeletal formation in vertebrates and to control both chondrocyte and osteoblast differentiation. In the present study, we analyzed the effect of Sonic hedgehog (Shh) on the osteoblastic and adipocytic commitment/differentiation. Recombinant N-terminal Shh (N-Shh) significantly increased the percentage of both the pluripotent mesenchymal cell lines C3H10T1/2 and ST2 and calvaria cells responding to bone morphogenetic protein 2 (BMP-2), in terms of osteoblast commitment as assessed by measuring alkaline phosphatase (ALP) activity. This synergistic effect was mediated, at least partly, through the positive modulation of the transcriptional output of BMPs via Smad signaling. Furthermore, N-Shh was found to abolish adipocytic differentiation of C3H10T1/2 cells both in the presence or absence of BMP-2. A short treatment with N-Shh was sufficient to dramatically reduce the levels of the adipocytic-related transcription factors C/EBPalpha and PPARgamma in both C3H10T1/2 and calvaria cell cultures. Given the inverse relationship between marrow adipocytes and osteoblasts with aging, agonists of the Hh signaling pathway might constitute potential drugs for preventing and/or treating osteopenic disorders. PMID- 11493645 TI - The non-catalytic domain of the Xenopus laevis auroraA kinase localises the protein to the centrosome. AB - Aurora kinases are involved in mitotic events that control chromosome segregation. All members of this kinase subfamily possess two distinct domains, a highly conserved catalytic domain and an N-terminal non-catalytic extension that varies in size and sequence. To investigate the role of this variable non catalytic region we overexpressed and purified Xenopus laevis auroraA (pEg2) histidine-tagged N-terminal peptide from bacterial cells. The peptide has no effect on the in vitro auroraA kinase activity, but it inhibits both bipolar spindle assembly and stability in Xenopus egg extracts. Unlike the full-length protein, the N-terminal domain shows only low affinity for paclitaxel-stabilised microtubules in vitro, but localises to the centrosomes in a microtubule dependent manner. When expressed in Xenopus XL2 cells, it is able to target the green fluorescent protein to centrosomes. Surprisingly, this is also true of the pEg2 catalytic domain, although to a lesser extent. The centrosome localisation of the N-terminal peptide was disrupted by nocodazole whereas localisation of the catalytic domain was not, suggesting that in order to efficiently localise to the centrosome, pEg2 kinase required the non-catalytic N-terminal domain and the presence of microtubules. PMID- 11493646 TI - trans-dominant inhibition of connexin-43 by mutant connexin-26: implications for dominant connexin disorders affecting epidermal differentiation. AB - Dominant mutations of GJB2-encoding connexin-26 (Cx26) have pleiotropic effects, causing either hearing impairment (HI) alone or in association with palmoplantar keratoderma (PPK/HI). We examined a British family with the latter phenotype and identified a new dominant GJB2 mutation predicted to eliminate the amino acid residue E42 (DeltaE42) in Cx26. To dissect the pathomechanisms that result in diverse phenotypes of dominant GJB2 mutations, we studied the effect of three Cx26 mutants (DeltaE42, D66H and R75W) identified in individuals with PPK/HI, and another (W44C) present in individuals with non-syndromic HI on gap junctional intercellular communication. We expressed mutant Cx26 alone and together with the epidermal connexins Cx26, Cx37 and Cx43 in paired Xenopus oocytes, and measured the intercellular coupling by dual voltage clamping. Homotypic expression of each connexin as well as co-expression of wild-type (wt) Cx26/wtCx43 and wtCx26/wtCx37 yielded variable, yet robust, levels of channel activity. However, all four Cx26 mutants were functionally impaired and failed to induce intercellular coupling. When co-expressed with wtCx26, all four mutants suppressed the wtCx26 channel activity consistent with a dominant inhibitory effect. However, only those Cx26 mutants associated with a skin phenotype also significantly (P<0.05) inhibited intercellular conductance of co-expressed wtCx43, indicating a direct interaction of mutant Cx26 units with wtCx43. These results demonstrate, for the first time, a trans-dominant negative effect of Cx26 mutants in vitro. Furthermore, they support a novel concept suggesting that the principal mechanism for manifestation of dominant GJB2 mutations in the skin is their dominant interference with the function of wtCx43. This assumption is further corroborated by our finding that Cx26 and Cx43 focally colocalize at gap junctional plaques in affected skin tissue of two carriers of DeltaE42. PMID- 11493647 TI - Retargeting of the mitochondrial protein p32/gC1Qr to a cytoplasmic compartment and the cell surface. AB - p32/gC1qR is a small acidic protein that has been reported to have a broad range of distinct functions and to associate with a wide array of cellular, viral and bacterial proteins. It has been found in each of the main cellular compartments including mitochondria, nucleus and cytoplasm and is also thought to be located at the plasma membrane and secreted into the extracellular matrix. The true physiological role(s) of p32 remains controversial because it has been difficult to reconcile all of the findings on protein interactions and the seemingly disparate observations on compartmentalisation. However, it has been proposed that p32 is somehow involved in transport processes connecting diverse cellular compartments and the cell surface. Here we show that native p32 appears to be localised mainly in the mitochondria and is not detectable on the cell surface. However, addition of a short tag to the N-terminus of p32 appears to block its mitochondrial targeting, resulting in redirection into a cytoplasmic vesicular pattern, overlapping with the endoplasmic reticulum. The redirection of p32 results in an alteration in and co-localisation with ER markers including calreticulin, a lumenal ER chaperone. Furthermore, we show both by immunofluorescence and cross-linking studies that this also results in cell surface expression of p32. These results indicate that, at least under certain circumstances, p32 can be retargeted and may help to provide an explanation for the diverse observations on its localization. PMID- 11493648 TI - MT1-MMP in rat testicular development and the control of Sertoli cell proMMP-2 activation. AB - Metalloproteases (MMPs) are likely to be involved in the restructuring events occurring in the testis throughout development. We here demonstrate that membrane type 1 (MT1)-MMP, a physiological activator of proMMP-2 under TIMP-2 control, is present within the testis together with MMP-2 and TIMP-2. In the prepubertal testis MT1-MMP immunoreactivity was uniformly distributed, whereas in the adult it was confined to the apical compartment of the tubules, where meiosis and spermiogenesis occur. We further showed that the two cell lineages (somatic and germinal) expressed MT1-MMP and TIMP-2, whereas MMP-2 was of somatic origin. To get a better picture into proMMP-2 activation, use was made of a model of cultured Sertoli cells treated with FSH or co-cultured with germ cells to mimic an immature or a mature developmental period, respectively. We found that follicle-stimulating hormone enhanced the expression of MMP-2 and TIMP-2 but not of MT1-MMP, and promoted the activation of proMMP-2. In co-cultures, a tremendous elevation and activation of MMP-2 was observed, which might relate to the processed MT1-MMP form solely detected in germ cells. That MMP-2 synthesis and activation are under local (germ cells) and hormonal (follicle-stimulating hormone) regulation emphasizes the importance of MMPs in testicular physiology. PMID- 11493649 TI - Fission yeast mfr1 activates APC and coordinates meiotic nuclear division with sporulation. AB - Meiosis is the developmental program by which sexually reproducing diploid organisms generate haploid gametes. In yeast, meiosis is followed by spore morphogenesis. These two events are normally coordinated in such a way that spore formation is dependent upon completion of the meiotic nuclear divisions. Here we describe a meiosis-specific protein, mfr1, that is involved in this coordination. mfr1 is an activator of the anaphase-promoting complex (APC), which is necessary for the rapid degradation of the cdc13 cyclin at the end of meiosis II, prior to the formation of spores. An mfr1 null mutant completes meiosis II but remains with high levels of cdc13 and cdc2 kinase activity and has considerably delayed spore formation. By analogy with the mitotic cell cycle, where proteolysis and inactivation of cdc2 kinase are necessary to trigger mitotic exit and cytokinesis, we propose that at the end of meiosis rapid and timely proteolysis of cyclins is required to switch on the differentiation program that eventually leads to the formation of haploid gametes. PMID- 11493650 TI - Priming of insulin granules for exocytosis by granular Cl(-) uptake and acidification. AB - ATP-dependent priming of the secretory granules precedes Ca(2+)-regulated neuroendocrine secretion, but the exact nature of this reaction is not fully established in all secretory cell types. We have further investigated this reaction in the insulin-secreting pancreatic B-cell and demonstrate that granular acidification driven by a V-type H(+)-ATPase in the granular membrane is a decisive step in priming. This requires simultaneous Cl(-) uptake through granular ClC-3 Cl(-) channels. Accordingly, granule acidification and priming are inhibited by agents that prevent transgranular Cl(-) fluxes, such as 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and an antibody against the ClC-3 channels, but accelerated by increases in the intracellular ATP:ADP ratio or addition of hypoglycemic sulfonylureas. We suggest that this might represent an important mechanism for metabolic regulation of Ca(2+)-dependent exocytosis that is also likely to be operational in other secretory cell types. PMID- 11493651 TI - Myosin II-dependent cylindrical protrusions induced by quinine in Dictyostelium: antagonizing effects of actin polymerization at the leading edge. AB - We found that amoeboid cells of Dictyostelium are induced by a millimolar concentration of quinine to form a rapidly elongating, cylindrical protrusion, which often led to sustained locomotion of the cells. Formation of the protrusion was initiated by fusion of a contractile vacuole with the cell membrane. During protrusion extension, a patch of the contractile vacuole membrane stayed undiffused on the leading edge of the protrusion for over 30 seconds. Protrusion formation was not inhibited by high osmolarity of the external medium (at least up to 400 mosM). By contrast, mutant cells lacking myosin II (mhc(-) cells) failed to extend protrusions upon exposure to quinine. When GFP-myosin-expressing cells were exposed to quinine, GFP-myosin was accumulated in the cell periphery forming a layer under the cell membrane, but a newly formed protrusion was initially devoid of a GFP-myosin layer, which gradually formed and extended from the base of the protrusion. F-actin was absent in the leading front of the protrusion during the period of its rapid elongation, and the formation of a layer of F-actin in the front was closely correlated with its slowing-down or retraction. Periodical or continuous detachment of the F-actin layer from the apical membrane of the protrusion, accompanied by a transient increase in the elongation speed at the site of detachment, was observed in some of the protrusions. The detached F-actin layers, which formed a spiral layer of F-actin in the case of continuous detachment, moved in the opposite direction of protrusion elongation. In the presence of both cytochalasin A and quinine, the protrusions formed were not cylindrical but spherical, which swallowed up the entire cellular contents. The estimated bulk flux into the expanding spherical protrusions of such cells was four-times higher than the flux into the elongating cylindrical protrusions of the cells treated with quinine alone. These results indicate that the force responsible for the quinine-induced protrusion is mainly due to contraction of the cell body, which requires normal myosin II functions, while actin polymerization is important in restricting the direction of its expansion. We will discuss the possible significance of tail contraction in cell movement in the multicellular phase of Dictyostelium development, where cell locomotion similar to that induced by quinine is often observed without quinine treatment, and in protrusion elongation in general. Movies available on-line PMID- 11493652 TI - c-Cbl ubiquitinates the EGF receptor at the plasma membrane and remains receptor associated throughout the endocytic route. AB - Cbl family members have an evolutionarily conserved role in attenuating receptor tyrosine kinase function. Their negative regulatory capacity depends on a Ring finger domain that interacts with ubiquitin conjugating enzymes. Cbl molecules constitute a novel type of E3 or ubiquitin ligase family that is recruited to phosphotyrosine motifs. Ubiquitination of the receptor system is coupled to its downregulation, but it is unclear at which point in the endocytic pathway Cbl molecules come into play. Using low temperature and a dynamin mutant, we find that c-Cbl associates with and ubiquitinates the activated epidermal growth factor (EGF) receptor at the plasma membrane in the absence of endocytosis. With the aid of confocal microscopy and immunogold electron microscopy, we could demonstrate that c-Cbl associates with the EGF receptor at the plasma membrane prior to receptor recruitment into clathrin-coated pits and remains associated throughout the clathrin-mediated endocytic pathway. c-Cbl and the EGF receptor also colocalize in internal vesicles of multivesicular endosomes. Our data are consistent with a role for c-Cbl in clathrin-mediated endocytosis of tyrosine kinase receptors, as well as their intracellular sorting. PMID- 11493653 TI - Store-operated influx of Ca(2+) in pancreatic beta-cells exhibits graded dependence on the filling of the endoplasmic reticulum. AB - The store-operated pathway for Ca(2+) entry was studied in individual mouse pancreatic beta-cells by measuring the cytoplasmic concentrations of Ca(2+) ([Ca(2+)](i)) and Mn(2+) ([Mn(2+)](i)) with the fluorescent indicator fura-2. Influx through the store-operated pathway was initially shut off by pre-exposure to 20 mM glucose, which maximally stimulates intracellular Ca(2+) sequestration. To avoid interference with voltage-dependent Ca(2+) entry the cells were hyperpolarized with diazoxide and the channel blocker methoxyverapamil was present. Activation of the store-operated pathway in response to Ca(2+) depletion of the endoplasmic reticulum was estimated from the sustained elevation of [Ca(2+)](i) or from the rate of increase in [Mn(2+)](i) due to influx of these extracellular ions. Increasing concentrations of the inositol 1,4,5-trisphosphate generating agonist carbachol or the sarco(endo)plasmatic reticulum Ca(2+)-ATPase inhibitor cyclopiazonic acid (CPA) cause gradual activation of the store-operated pathway. In addition, the carbachol- and CPA-induced influx of Mn(2+) depended on store filling in a graded manner. The store-operated influx of Ca(2+)/Mn(2+) was inhibited by Gd(3+) and 2-aminoethoxydiphenyl borate but neither of these agents discriminated between store-operated and voltage-dependent entry. The finely tuned regulation of the store-operated mechanisms in the beta-cell has direct implications for the control of membrane potential and insulin secretion. PMID- 11493654 TI - SHP-2 complex formation with the SHP-2 substrate-1 during C2C12 myogenesis. AB - Myogenesis is a highly ordered process that involves the expression of muscle specific genes, cell-cell recognition and multinucleated myotube formation. Although protein tyrosine kinases have figured prominently in myogenesis, the involvement of tyrosine phosphatases in this process is unknown. SHP-2 is an SH2 domain-containing tyrosine phosphatase, which positively regulates growth and differentiation. We show that in C2C12 myoblasts, SHP-2 becomes upregulated early on during myogenesis and associates with a 120 kDa tyrosyl-phosphorylated complex. We have identified that the 120 kDa complex consists of the SHP-2 substrate-1 (SHPS-1) and the Grb2-associated binder-1 (Gab-1). SHPS-1, but not Gab-1, undergoes tyrosyl phosphorylation and association with SHP-2 during myogenesis, the kinetics of which correlate with the expression of MyoD. Either constitutive expression or inducible activation of MyoD in 10T(1/2) fibroblasts promotes SHPS-1 tyrosyl phosphorylation and its association with SHP-2. It has been shown that p38 mitogen-activated protein kinase (MAPK) activity is required for the expression/activation of MyoD and MyoD-responsive genes. Inhibition of p38 MAPK by SB203580 in differentiating C2C12 myoblasts blocks MyoD expression, SHPS-1 tyrosyl phosphorylation and the association of SHPS-1 with SHP-2. These data suggest that SHPS-1/SHP-2 complex formation is an integral signaling component of skeletal muscle differentiation. PMID- 11493655 TI - Peroxisomal membrane proteins are properly targeted to peroxisomes in the absence of COPI- and COPII-mediated vesicular transport. AB - The classic model for peroxisome biogenesis states that new peroxisomes arise by the fission of pre-existing ones and that peroxisomal matrix and membrane proteins are recruited directly from the cytosol. Recent studies challenge this model and suggest that some peroxisomal membrane proteins might traffic via the endoplasmic reticulum to peroxisomes. We have studied the trafficking in human fibroblasts of three peroxisomal membrane proteins, Pex2p, Pex3p and Pex16p, all of which have been suggested to transit the endoplasmic reticulum before arriving in peroxisomes. Here, we show that targeting of these peroxisomal membrane proteins is not affected by inhibitors of COPI and COPII that block vesicle transport in the early secretory pathway. Moreover, we have obtained no evidence for the presence of these peroxisomal membrane proteins in compartments other than peroxisomes and demonstrate that COPI and COPII inhibitors do not affect peroxisome morphology or integrity. Together, these data fail to provide any evidence for a role of the endoplasmic reticulum in peroxisome biogenesis. PMID- 11493656 TI - Life in the stone age. By Caveman. PMID- 11493657 TI - Phosphatidylinositol and inositol phosphate metabolism. PMID- 11493661 TI - The organisation and functions of local Ca(2+) signals. AB - Calcium (Ca(2+)) is a ubiquitous intracellular messenger, controlling a diverse range of cellular processes, such as gene transcription, muscle contraction and cell proliferation. The ability of a simple ion such as Ca(2+) to play a pivotal role in cell biology results from the facility that cells have to shape Ca(2+) signals in space, time and amplitude. To generate and interpret the variety of observed Ca(2+) signals, different cell types employ components selected from a Ca(2+) signalling 'toolkit', which comprises an array of homeostatic and sensory mechanisms. By mixing and matching components from the toolkit, cells can obtain Ca(2+) signals that suit their physiology. Recent studies have demonstrated the importance of local Ca(2+) signals in defining the specificity of the interaction of Ca(2+) with its targets. Furthermore, local Ca(2+) signals are the triggers and building blocks for larger global signals that propagate throughout cells. PMID- 11493662 TI - Mechanisms of capacitative calcium entry. AB - Capacitative Ca(2+) entry involves the regulation of plasma membrane Ca(2+) channels by the filling state of intracellular Ca(2+) stores in the endoplasmic reticulum (ER). Several theories have been advanced regarding the mechanism by which the stores communicate with the plasma membrane. One such mechanism, supported by recent findings, is conformational coupling: inositol 1,4,5 trisphosphate (Ins(1,4,5)P(3)) receptors in the ER may sense the fall in Ca(2+) levels through Ca(2+)-binding sites on their lumenal domains, and convey this conformational information directly by physically interacting with Ca(2+) channels in the plasma membrane. In support of this idea, in some cell types, store-operated channels in excised membrane patches appear to depend on the presence of both Ins(1,4,5)P(3) and Ins(1,4,5)P(3) receptors for activity; in addition, inhibitors of Ins(1,4,5)P(3) production that either block phospholipase C or inhibit phosphatidylinositol 4-kinase can block capacitative Ca(2+) entry. However, the electrophysiological current underlying capacitative Ca(2+) entry is not blocked by an Ins(1,4,5)P(3) receptor antagonist, and the blocking effects of a phospholipase C inhibitor are not reversed by the intracellular application of Ins(1,4,5)P(3). Furthermore, cells whose Ins(1,4,5)P(3) receptor genes have been disrupted can nevertheless maintain their capability to activate capacitative Ca(2+) entry channels in response to store depletion. A tentative conclusion is that multiple mechanisms for signaling capacitative Ca(2+) entry may exist, and involve conformational coupling in some cell types and perhaps a diffusible signal in others. PMID- 11493663 TI - Three dimensional configuration of the secretory pathway and segregation of secretion granules in the yeast Saccharomyces cerevisiae. AB - The structural elements of the secretory pathway in the budding yeast Saccharomyces cerevisiae were analyzed by 3D stereo-electron microscopy using relatively thick sections in which membranes were selectively impregnated. In a wild-type strain, tubular networks of various sizes and staining properties were distributed throughout the cytoplasm. As a rule, wide-meshed, lightly stained polygonal networks were connected to more or less fenestrated sheets of endoplasmic reticulum (ER). Some of these networks were continuous with more intensely stained networks and narrower meshes that displayed at their intersections nodular dilations that progressively increased in size and staining properties to reach those of secretion granules. Such networks presumably corresponded to Golgi elements. Indeed, stacked cisternae typical of the mammalian Golgi apparatus are rarely found in wild-type cells. However, if it is assumed that the Golgi apparatus plays a key role in the segregation and maturation of secretion granules, then tubular networks with nodular dilations should be equivalent to parts of this organelle. In correlation with the increase in size and density of the nodules there was a decrease in diameter and staining intensity of the interconnecting tubules. These results parallel observations on the formation of secretory granules in mammalian cells and suggest that the segregation of secretory material is concomitant with the progressive perforation and tubulization of previously unperforated sheets. When the sec21-3 thermosensitive mutant was examined at the nonpermissive temperature (37 degrees C), the secretory pathway was blocked at exit from the ER, which started to accumulate as clusters of narrow, anastomosed, unperforated ribbon-like elements. When the block was released by shifting down to permissive temperature (24 degrees C), tubular networks of various sizes and caliber, presumably Golgi in nature, formed as soon as 5 minutes after release of the block. At later time intervals, granules of various sizes and densities appeared to be released by rupture of these tubular networks or even to form at the edges of ER fenestrae. These observations support a dynamic maturation process in which the formation of secretion granules occurs by means of an oriented series of membrane transformations starting at the ER and culminating with the liberation of secretion granules from Golgi networks. PMID- 11493664 TI - The ARF exchange factors Gea1p and Gea2p regulate Golgi structure and function in yeast. AB - The Sec7 domain guanine nucleotide exchange factors (GEFs) for the GTPase ARF are highly conserved regulators of membrane dynamics. Their precise molecular roles in different trafficking steps within the cell have not been elucidated. We present a functional analysis of two members of this family, Gea1p and Gea2p, in the yeast Saccharomyces cerevisiae. Gea1p and Gea2p can functionally replace each other, but at least one is necessary for viability. Temperature sensitive gea mutants were generated and found to have defects in ER-Golgi and intra-Golgi transport. Similar to mutants in COPI subunits in yeast, gea mutants had a cargo selective secretion defect, in that some proteins continued to be secreted whereas others were blocked in the ER or early Golgi. Like yeast arf mutants, the rate of transport of those proteins that continued to be secreted was slowed. In addition, the structure of Golgi elements was severely perturbed in gea mutants. We conclude that Gea1p and Gea2p play an important role in the structure and functioning of the Golgi apparatus in yeast. PMID- 11493665 TI - FYVE and coiled-coil domains determine the specific localisation of Hrs to early endosomes. AB - Hrs, an essential tyrosine kinase substrate, has been implicated in intracellular trafficking and signal transduction pathways. The protein contains several distinctive domains, including an N-terminal VHS domain, a phosphatidylinositol 3 phosphate (PtdIns(3)P)-binding FYVE domain and two coiled-coil domains. Here we have investigated the roles of these domains in the subcellular localisation of Hrs. Hrs was found to colocalise extensively with EEA1, an established marker of early endosomes. While the membrane association of EEA1 was abolished in the presence of a dominant negative mutant of the endosomal GTPase Rab5, the localisation of Hrs to early endosomes was Rab5 independent. The VHS-domain was nonessential for the subcellular targeting of Hrs. In contrast, the FYVE domain as well as the second coiled-coil domain, which has been shown to bind to SNAP 25, were required for targeting of Hrs to early endosomes. A small construct consisting of only these two domains was correctly localised to early endosomes, whereas a point mutation (R183A) in the PtdIns(3)P-binding pocket of the FYVE domain inhibited the membrane targeting of Hrs. Thus, like EEA1, the endosomal targeting of Hrs is mediated by a PtdIns(3)P-binding FYVE domain in cooperation with an additional domain. We speculate that binding to PtdIns(3)P and a SNAP-25 related molecule may target Hrs specifically to early endosomes. PMID- 11493666 TI - Assembly of C. elegans apical junctions involves positioning and compaction by LET-413 and protein aggregation by the MAGUK protein DLG-1. AB - Specialised subapical junctions play a critical role in maintaining epithelial cell polarity and tissue integrity, and provide a platform for intracellular signalling. Here we analyse the roles of C. elegans genes let-413 and dlg-1, a homologue of Drosophila lethal discs large, in the assembly of the C. elegans apical junction (CeAJ), and provide the first characterisation of this structure. We have identified dlg-1 as an essential gene in an RNA interference screen against C. elegans homologues of genes encoding proteins involved in tight or septate junction formation. We show that DLG-1 colocalises with the junctional protein JAM-1 at CeAJs in a unit distinct from HMP-1/alpha-catenin, and apical to the laterally localised LET-413. Loss of dlg-1 activity leads to JAM-1 mislocalisation and the disappearance of the electron-dense component of the CeAJs, but only mild adhesion and polarity defects. In contrast, loss of let-413 activity leads to the formation of basally extended discontinuous CeAJs and strong adhesion and polarity defects. Interestingly, in LET-413-deficient embryos, CeAJ markers are localised along the lateral membrane in a manner resembling that observed in wild-type embryos at the onset of epithelial differentiation. We conclude that the primary function of LET-413 is to correctly position CeAJ components at a discrete subapical position. Furthermore, we propose that DLG-1 is required to aggregate JAM-1 and other proteins forming the electron-dense CeAJ structure. Our data suggest that epithelial adhesion is maintained by several redundant systems in C. elegans. PMID- 11493667 TI - pp60(c-src) and related tyrosine kinases: a role in the assembly and reorganization of matrix adhesions. AB - Activation of tyrosine kinases during integrin-mediated cell-matrix adhesion is involved both in the regulation of focal contact assembly and in the initiation of signaling processes at the cell-matrix adhesive interface. In order to determine the role of pp60(c-src) and related kinases in these processes, we have compared the dynamic reorganization of phosphotyrosine, vinculin, focal adhesion kinase and tensin in cells with altered expression of Src-family kinases. Both null cells for pp60(c-src) and triple knockout cells for pp60(c-src), pp59(fyn), and pp62(c-yes) exhibited decreased phosphotyrosine levels in focal contacts when compared with wild-type cells. pp60(c-src)-null cells also exhibited faster assembly of cell-matrix adhesions and a more exuberant recruitment of FAK to these sites. Tensin, which normally segregates into fibrillar adhesions was localized in large focal contacts in the two mutant cell lines, suggesting involvement of pp60(c-src) in the segregation of focal contacts and fibrillar adhesions. Moreover, treatment of wild-type cells with tyrphostin AG1007, which inhibits both pp60(c-src) and FAK activity, induced accumulation of tensin in peripheral focal adhesions. These findings demonstrate that Src family kinases, and pp60(c-src) in particular, have a central role in regulating protein dynamics at cell-matrix interfaces, both during early stages of interaction and in mature focal contacts. PMID- 11493668 TI - Functional interaction between the SSeCKS scaffolding protein and the cytoplasmic domain of beta1,4-galactosyltransferase. AB - The beta1,4-galactosyltransferase family contains at least seven unique gene products, of which beta1,4-galactosyltransferase I (GalT) is the most exhaustively studied. GalT exists in the Golgi complex, similar to many other glycosyltransferases, as well as on the cell surface, where it functions as a signaling receptor for extracellular glycoside ligands. When expressed on the surface, GalT associates with the cytoskeleton and, upon ligand-induced aggregation, induces cell-type specific intracellular signal cascades. In an effort to define the mechanisms by which surface GalT exerts these intracellular effects, we used the yeast two-hybrid system to identify proteins that specifically interact with the GalT cytoplasmic domain. The yeast two-hybrid screen identified two distinct clones (1.12 and 2.52) that showed identity to portions of SSeCKS (Src Suppressed C Kinase Substrate). SSeCKS is a previously defined kinase and cytoskeleton scaffolding protein whose subcellular distribution and functions are remarkably similar to those attributed to GalT. Both SSeCKS and GalT have been localized to the perinuclear/Golgi region as well as to filopodia/lamellipodia. SSeCKS and GalT have been implicated in regulating cell growth, actin filament dynamics, and cell spreading. Interestingly, 1.12 and 2.52-GFP constructs were localized to subcellular domains that correlated with the two purported subcellular distributions for GalT; 2.52 being confined to the Golgi, whereas 1.12 localized primarily to filopodia. Coimmunoprecipitation assays demonstrate stable binding between the GalT cytoplasmic domain and the 1.12 and 2.52 domains of SSeCKS in appropriately transfected cells. Similar assays demonstrate binding between the endogenous GalT and SSeCKS proteins also. Coimmunoprecipitation assays were performed in both directions and produced similar results (i.e. using either anti-GalT domain or anti-SSeCKS domain antibodies as the precipitating reagent). A functional interaction between the GalT cytoplasmic domain and SSeCKS was illustrated by the ability of either the 1.12 or 2.52 SSeCKS domain to restore a normal adhesive phenotype in cells overexpressing the TL-GFP dominant negative construct. TL-GFP is composed of the GalT cytoplasmic and transmembrane domains fused to GFP, and leads to a loss of cell adhesion on laminin by displacing the endogenous GalT from its cytoskeleton binding sites. This is the first reported interaction between a glycosyltransferase and a scaffolding protein, and suggests that SSeCKS serve to integrate the various functions ascribed to the GalT cytoplasmic domain. PMID- 11493669 TI - Quantitative analysis of the EGF receptor autocrine system reveals cryptic regulation of cell response by ligand capture. AB - Autocrine signaling is important in normal tissue physiology as well as pathological conditions. It is difficult to analyze these systems, however, because they are both self-contained and recursive. To understand how parameters such as ligand production and receptor expression influence autocrine activity, we investigated a human epidermal growth factor/epidermal growth factor receptor (EGF/EGFR) loop engineered into mouse B82 fibroblasts. We varied the level of ligand production using the tet-off expression system and used metalloprotease inhibitors to modulate ligand release. Receptor expression was varied using antagonistic blocking antibodies. We compared autocrine ligand release with receptor activation using a microphysiometer-based assay and analyzed our data using a quantitative model of ligand release and receptor dynamics. We found that the activity of our autocrine system could be described in terms of a simple ratio between the rate of ligand production (V(LT)) and the rate of receptor production (V(R)). At a V(LT)/V(R) ratio of <0.3, essentially no ligand was found in the extracellular medium, but a significant number of cell receptors (30-40%) were occupied. As the V(LT)/V(R) ratio increased from 0.3 towards unity, receptor occupancy increased and significant amounts of ligand appeared in the medium. Above a V(LT)/V(R) ratio of 1.0, receptor occupancy approached saturation and most of the released ligand was lost into the medium. Analysis of human mammary epithelial cells showed that a V(LT)/V(R) ratio of < 5 x 10(-4)was sufficient to evoke >20% of a maximal proliferative response. This demonstrates that natural autocrine systems can be active even when no ligand appears in the extracellular medium. PMID- 11493670 TI - Chondroitin sulfate and cytoplasmic domain-dependent membrane targeting of the NG2 proteoglycan promotes retraction fiber formation and cell polarization. AB - Targeting of the NG2 proteoglycan to cellular retraction fibers was studied by expressing mutant NG2 molecules lacking specific structural elements of the proteoglycan. Both the cytoplasmic domain and the chondroitin sulfate chain of NG2 appear to have roles in sorting NG2 to subcellular microdomains destined to become retraction fibers. Neither of these structural features alone is sufficient to allow optimal targeting of NG2 to retraction fibers, but together they promote efficient localization of the proteoglycan to these sites. This pattern of NG2 sorting seems to be necessary for optimal retraction fiber formation, as cells expressing poorly targeted NG2 mutants are noticeably deficient in their ability to extend retraction fibers. Furthermore, retraction fiber formation correlates strongly with the tendency of cells to assume a polarized morphology with NG2-positive retraction fibers at one pole of the cell and actin-rich lamellipodia at the other. This polarization can be triggered either through engagement of NG2 by the substratum or by exposure to lysophosphatidic acid, a potent activator of the rho GTPase. These results suggest a possible role for NG2 in regulating rho-dependent mechanisms in the trailing processes of motile cells. PMID- 11493671 TI - Nuclear translocation of ferritin in corneal epithelial cells. AB - Our previous studies have shown that ferritin within developing avian corneal epithelial cells is predominantly a nuclear protein and that one function of the molecule in this location is to protect DNA from UV damage. To elucidate the mechanism for this tissue-specific nuclear translocation, cultured corneal epithelial cells and corneal fibroblasts were transfected with a series of deletion constructs for the heavy chain of ferritin, ferritin-H, tagged with a human c-myc epitope. The subcellular localization of the ferritin was determined by immunofluorescence for the myc-tag. For the corneal epithelial cells, the first 10 or the last 30 amino acids of ferritin-H could be deleted without affecting the nuclear localization. However, larger deletions of these areas, or deletions along the length of the body of the molecule, resulted largely in retention of the truncated proteins within the cytoplasm. Thus, it seems that no specific region functions as an NLS. Immunoblotting analysis of SDS-PAGE separated extracts suggests that assembly of the supramolecular form of ferritin is not necessary for successful nuclear translocation, because one deletion construct that failed to undergo supramolecular assembly showed nuclear localization. In transfected fibroblasts, the endogenous ferritin remained predominantly in the cytoplasm, as did that synthesized from transfected full length ferritin constructs and from two deletion constructs encoding truncated chains that could still assemble into the supramolecular form of ferritin. However, those truncated chains that were unable to participate in supramolecular assembly generally showed both nuclear and cytoplasmic localization, indicating that, in this cell type, supramolecular assembly is involved in restricting ferritin to the cytoplasm. These data suggest that for corneal epithelial cells, the nuclear localization of ferritin most likely involves a tissue-specific mechanism that facilitates transport into the nucleus, whereas, in fibroblasts, the cytoplasmic retention involves supramolecular assembly that prevents passive diffusion into the nucleus. PMID- 11493672 TI - Inhibition of melanogenesis in response to oxidative stress: transient downregulation of melanocyte differentiation markers and possible involvement of microphthalmia transcription factor. AB - H(2)O(2) and other reactive oxygen species are key regulators of many intracellular pathways. Within mammalian skin, H(2)O(2) is formed as a byproduct of melanin synthesis, and following u.v. irradiation. We therefore analyzed its effects on melanin synthesis. The activity of the rate-limiting melanogenic enzyme, tyrosinase, decreased in H(2)O(2)-treated mouse and human melanoma cells. This inhibition was concentration- and time-dependent in the B16 melanoma model. Maximal inhibition (50-75%) occurred 8-16 hours after a 20 minute exposure to 0.5 mM H(2)O(2). B16 cells withstand this treatment adequately, as shown by a small effect on glutathione levels and a rapid recovery of basal lipid peroxidation levels. Enzyme activities also recovered, beginning to increase 16-20 hours after the treatment. Inhibition of enzyme activities reflected decreased protein levels. mRNAs for tyrosinase, tyrosinase-related protein 1, dopachrome tautomerase, silver protein and melanocortin 1 receptor also decreased after H(2)O(2) treatment, and recovered at different rates. Downregulation of melanocyte differentiation markers mRNAs was preceded by a decrease in microphthalmia transcription factor (Mitf) gene expression, which was quantitatively similar to the decrease achieved using 12-O-tetradecanoylphorbol 13-acetate. Recovery of basal Mitf mRNA levels was also observed clearly before that of tyrosinase. Therefore, oxidative stress may lead to hypopigmentation by mechanisms that include a microphthalmia-dependent downregulation of the melanogenic enzymes. PMID- 11493673 TI - The Bub2-dependent mitotic pathway in yeast acts every cell cycle and regulates cytokinesis. AB - In eukaryotes an abnormal spindle activates a conserved checkpoint consisting of the MAD and BUB genes that results in mitotic arrest at metaphase. Recently, we and others identified a novel Bub2-dependent branch to this checkpoint that blocks mitotic exit. This cell-cycle arrest depends upon inhibition of the G protein Tem1 that appears to be regulated by Bfa1/Bub2, a two-component GTPase activating protein, and the exchange factor Lte1. Here, we find that Bub2 and Bfa1 physically associate across the entire cell cycle and bind to Tem1 during mitosis and early G1. Bfa1 is multiply phosphorylated in a cell-cycle-dependent manner with the major phosphorylation occurring in mitosis. This Bfa1 phosphorylation is Bub2-dependent. Cdc5, but not Cdc15 or Dbf2, partly controls the phosphorylation of Bfa1 and also Lte1. Following spindle checkpoint activation, the cell cycle phosphorylation of Bfa1 and Lte1 is protracted and some species are accentuated. Thus, the Bub2-dependent pathway is active every cell cycle and the effect of spindle damage is simply to protract its normal function. Indeed, function of the Bub2 pathway is also prolonged during metaphase arrests imposed by means other than checkpoint activation. In metaphase cells Bub2 is crucial to restrain downstream events such as actin ring formation, emphasising the importance of the Bub2 pathway in the regulation of cytokinesis. Our data is consistent with Bub2/Bfa1 being a rate-limiting negative regulator of downstream events during metaphase. PMID- 11493674 TI - A novel mammalian receptor for the evolutionarily conserved type II GnRH. AB - Mammalian gonadotropin-releasing hormone (GnRH I: pGlu-His-Trp-Ser-Tyr-Gly-Leu Arg-Pro-Gly-NH2) stimulates pituitary gonadotropin secretion, which in turn stimulates the gonads. Whereas a hypothalamic form of GnRH of variable structure (designated type I) had been shown to regulate reproduction through a cognate type I receptor, it has recently become evident that most vertebrates have one or two other forms of GnRH. One of these, designated type II GnRH (GnRH II: pGlu-His Ser-His-Gly-Trp-Tyr-Pro-Gly-NH2), is conserved from fish to man and is widely distributed in the brain, suggesting important neuromodulatory functions such as regulating K+ channels and stimulating sexual arousal. We now report the cloning of a type II GnRH receptor from marmoset cDNA. The receptor has only 41% identity with the type I receptor and, unlike the type I receptor, has a carboxyl-terminal tail. The receptor is highly selective for GnRH II. As with the type I receptor, it couples to G(alpha)q/11 and also activates extracellular signal-regulated kinase (ERK1/2) but differs in activating p38 mitogen activated protein (MAP) kinase. The type II receptor is more widely distributed than the type I receptor and is expressed throughout the brain, including areas associated with sexual arousal, and in diverse non-neural and reproductive tissues, suggesting a variety of functions. Surprisingly, the type II receptor is expressed in the majority of gonadotropes. The presence of two GnRH receptors in gonadotropes, together with the differences in their signaling, suggests different roles in gonadotrope functioning. PMID- 11493675 TI - Cleavage of the human respiratory syncytial virus fusion protein at two distinct sites is required for activation of membrane fusion. AB - Preparations of purified full-length fusion (F) protein of human respiratory syncytial virus (HRSV) expressed in recombinant vaccinia-F infected cells, or of an anchorless mutant (F(TM(-))) lacking the C-terminal 50 amino acids secreted from vaccinia-F(TM(-))-infected cells contain a minor polypeptide that is an intermediate product of proteolytic processing of the F protein precursor F0. N terminal sequencing of the intermediate demonstrated that it is generated by cleavage at a furin-motif, residues 106-109 of the F sequence. By contrast, the F1 N terminus derives from cleavage at residue 137 of F0 which is also C-terminal to a furin recognition site at residues 131-136. Site-directed mutagenesis indicates that processing of F0 protein involves independent cleavage at both sites. Both cleavages are required for the F protein to be active in membrane fusion as judged by syncytia formation, and they allow changes in F structure from cone- to lollipop-shaped spikes and the formation of rosettes by anchorless F. PMID- 11493676 TI - Theoretical emission line ratios for [Fe III] and [Fe VII] applicable to the optical and infrared spectra of gaseous nebulae. AB - Recent calculations of electron impact excitation rates and Einstein A coefficients for transitions among the 3d(6) levels of Fe III and among the 3d(2) levels of Fe VII are used to derive theoretical emission line ratios applicable to the optical and infrared spectra of gaseous nebulae. Results for [Fe III] are generated for electron temperatures T(e) = 7,000-20,000 K and densities N(e) = 10(2)-10(8) cm(-3), whereas those for [Fe VII] are provided for T(e) = 10,000 30,000 K and N(e) = 10(2)-10(8) cm(-3). The theoretical line ratios are significantly different in some instances from earlier calculations and resolve discrepancies between theory and observation found for the planetary nebulae IC 4997 and NGC 7027. PMID- 11493677 TI - Transgene analysis proves mRNA trans-splicing at the complex mod(mdg4) locus in Drosophila. AB - The Drosophila BTB domain containing gene mod(mdg4) produces a large number of protein isoforms combining a common N-terminal region of 402 aa with different C termini. We have deduced the genomic structure of this complex locus and found that at least seven of the mod(mdg4) isoforms are encoded on both of its antiparallel DNA strands, suggesting the generation of mature mRNAs by trans splicing. In transgenic assays, we demonstrate the ability of Drosophila to produce mod(mdg4) mRNAs by trans-splicing of pre-mRNAs generated from transgenes inserted at distant chromosomal positions. Furthermore, evidence is presented for occurring of trans-splicing of mod(mdg4)-specific exons encoded by the parallel DNA strand. The mod(mdg4) locus represents a new type of complex gene structure in which genetic complexity is resolved by extensive trans-splicing, giving important implications for genome sequencing projects. Demonstration of naturally occurring trans-splicing in the model organism Drosophila opens new experimental approaches toward an analysis of the underlying mechanisms. PMID- 11493678 TI - Heart-targeted overexpression of caspase3 in mice increases infarct size and depresses cardiac function. AB - Up-regulation of proapoptotic genes has been reported in heart failure and myocardial infarction. To determine whether caspase genes can affect cardiac function, a transgenic mouse was generated. Cardiac tissue-specific overexpression of the proapoptotic gene Caspase3 was induced by using the rat promoter of alpha-myosin heavy chain, a model that may represent a unique tool for investigating new molecules and antiapoptotic therapeutic strategies. Cardiac specific Caspase3 expression induced transient depression of cardiac function and abnormal nuclear and myofibrillar ultrastructural damage. When subjected to myocardial ischemia-reperfusion injury, Caspase3 transgenic mice showed increased infarct size and a pronounced susceptibility to die. In this report, we document an unexpected property of the proapoptotic gene caspase3 on cardiac contractility. Despite inducing ultrastructural damage, Caspase3 does not trigger a full apoptotic response in the cardiomyocyte. We also implicate Caspase3 in determining myocardial infarct size after ischemia-reperfusion injury, because its cardiomyocyte-specific overexpression increases infarct size. PMID- 11493680 TI - The active site dynamics of 4-chlorobenzoyl-CoA dehalogenase. AB - A molecular dynamics study was performed to compare the differences in the active site dynamics of the wild-type and W137F mutant enzymes of 4-chlorobenzoyl-CoA dehalogenase. Only in the wild-type simulation are conformations formed between the catalytic Asp-145 and 4-chlorobenzoyl-CoA, which resemble the ab initio calculated gas-phase transition-state geometry. In the W137F simulation, the hydrogen bond formed between His-90 and Asp-145 persisted throughout the simulation, causing the carboxylate of Asp-145 to be distant from the benzoyl ring of 4-chlorobenzoyl-CoA. In both simulations, water molecules were able to diffuse into the active site of the enzymes. The trajectories provide insight into the routes that water may use to get into position for the hydrolysis portion of the dehalogenation reaction. In both simulations, the water molecule entering the active site forms a hydrogen bond with Asp-145. PMID- 11493679 TI - The yeast mutant vps5Delta affected in the recycling of Golgi membrane proteins displays an enhanced vacuolar Mg2+/H+ exchange activity. AB - Growth of the yeast vacuolar protein-sorting mutant vps5Delta affected in the endosome-to-Golgi retromer complex was more sensitive to Mg2+-limiting conditions than was the growth of the wild-type (WT) strain. This sensitivity was enhanced at acidic pH. The vps5Delta strain was also sensitive to Al3+, known to inhibit Mg2+ uptake in yeast cells. In contrast, it was found to be resistant to Ni2+ and Co2+, two cytotoxic analogs of Mg2+. Resistance to Ni2+ did not seem to result from the alteration of plasma-membrane transport properties because mutant and WT cells displayed similar Ni2+ uptake. After plasma-membrane permeabilization, intracellular Ni2+ uptake in vps5Delta cells was 3-fold higher than in WT cells, which is consistent with the implication of the vacuole in the observed phenotypes. In reconstituted vacuolar vesicles prepared from vps5Delta, the rates of H+ exchange with Ni2+, Co2+, and Mg2+ were increased (relative to WT) by 170%, 130%, and 50%, respectively. The rates of H+ exchange with Ca2+, Cd2+, and K+ were similar in both strains, as were alpha-mannosidase and H+-ATPase activities, and SDS/PAGE patterns of vacuolar proteins. Among 14 other vacuolar protein sorting mutants tested, only the 8 mutants affected in the recycling of trans Golgi network membrane proteins shared the same Ni2+ resistance phenotype as vps5Delta. It is proposed that a trans-Golgi network Mg2+/H+ exchanger, mislocalized to vps5Delta vacuole, could be responsible for the phenotypes observed in vivo and in vitro. PMID- 11493681 TI - Phase transition between disordered and ordered foraging in Pharaoh's ants. AB - The complex collective behavior seen in many insect societies strongly suggests that a minimum number of workers are required for these societies to function effectively. Here we investigated the transition between disordered and ordered foraging in the Pharaoh's ant. We show that small colonies forage in a disorganized manner, with a transition to organized pheromone-based foraging in larger colonies. We also show that when food sources are difficult to locate through independent searching, this transition is first-order and exhibits hysteresis, comparable to a first-order phase transition found in many physical systems. To our knowledge, this is the first experimental evidence of a behavioral phase transition between a maladaptive (disorganized) and an adaptive (organized) state. PMID- 11493682 TI - Ubiquitination and degradation of Syk and ZAP-70 protein tyrosine kinases in human NK cells upon CD16 engagement. AB - Syk and ZAP-70 nonreceptor protein tyrosine kinases (PTKs) are essential elements in several cascades coupling immune receptors to intracellular responses. The critical role of these kinases in promoting the propagation of intracellular signaling requires a tight regulation of their activity, thus the existence of a negative feedback loop regulating their expression can be hypothesized. Herein, we have investigated whether ubiquitin-dependent proteolysis could be a mechanism responsible for controlling the fate of Syk and ZAP-70 after their immunoreceptor induced activation. We found that both Syk and ZAP-70 become ubiquitinated in response to aggregation of the low affinity Fc receptor for IgG (CD16) on human natural killer cells. We confirmed the identity of the major in vivo ubiquitinated kinase species by performing an in vitro ubiquitination assay. In addition, we found that after CD16 stimulation, ubiquitinated forms of Syk and ZAP-70 associate with the receptor complex. After CD16 engagement, we also observed a decrease in the stability of Syk and ZAP-70 PTKs that is counteracted by pretreatment with either proteasome or lysosomal inhibitors. Moreover, in the presence of the proteasome inhibitor, epoxomicin, we observed an accumulation of ubiquitinated forms of both kinases. Our findings provide evidence of ligand induced ubiquitination of nonreceptor PTKs belonging to the Syk family and propose the ubiquitin-dependent proteasome-mediated degradation pathway as a mechanism for attenuating the propagation of intracellular signaling initiated by immune receptor engagement. PMID- 11493683 TI - Highly selective water channel activity measured by voltage clamp: analysis of planar lipid bilayers reconstituted with purified AqpZ. AB - Aquaporins are membrane channels selectively permeated by water or water plus glycerol. Conflicting reports have described ion conductance associated with some water channels, raising the question of whether ion conductance is a general property of the aquaporin family. To clarify this question, a defined system was developed to simultaneously measure water permeability and ion conductance. The Escherichia coli water channel aquaporin-Z (AqpZ) was studied, because it is a highly stable tetramer. Planar lipid bilayers were formed from unilamellar vesicles containing purified AqpZ. The hydraulic conductivity of bilayers made from the total extract of E. coli lipids increased 3-fold if reconstituted with AqpZ, but electric conductance was unchanged. No channel activity was detected under voltage-clamp conditions, indicating that less than one in 10(9) transport events is electrogenic. Microelectrode measurements were simultaneously undertaken adjacent to the membrane. Changes in sodium concentration profiles accompanying transmembrane water flow permitted calculation of the activation energies: 14 kcal/mol for protein-free lipid bilayers and 4 kcal/mol for lipid bilayers containing AqpZ. Neither the water permeability nor the electric conductivity exhibited voltage dependence. This sensitive system demonstrated that AqpZ is permeated by water but not charged ions and should permit direct analyses of putative electrogenic properties of other aquaporins. PMID- 11493684 TI - Cross talk among calcineurin, Sp1/Sp3, and NFAT in control of p21(WAF1/CIP1) expression in keratinocyte differentiation. AB - Calcium functions as a trigger for the switch between epithelial cell growth and differentiation. We report here that the calcium/calmodulin-dependent phosphatase calcineurin is involved in this process. Treatment of primary mouse keratinocytes with cyclosporin A, an inhibitor of calcineurin activity, suppresses the expression of terminal differentiation markers and of p21(WAF1/Cip1) and p27(KIP1), two cyclin-dependent kinase inhibitors that are usually induced with differentiation. In parallel with down-modulation of the endogenous genes, suppression of calcineurin function blocks induction of the promoters for the p21(WAF1/Cip1) and loricrin differentiation marker genes, whereas activity of these promoters is enhanced by calcineurin overexpression. The calcineurin- responsive region of the p21 promoter maps to a 78-bp Sp1/Sp3-binding sequence next to the TATA box, and calcineurin induces activity of the p21 promoter through Sp1/Sp3-dependent transcription. We find that the endogenous NFAT-1 and 2 transcription factors, major downstream targets of calcineurin, associate with Sp1 in keratinocytes in a calcineurin-dependent manner, and calcineurin up regulates Sp1/Sp3-dependent transcription and p21 promoter activity in synergism with NFAT1/2. Thus, our study reveals an important role for calcineurin in control of keratinocyte differentiation and p21 expression, and points to a so far-unsuspected interconnection among this phosphatase, NFATs, and Sp1/Sp3 dependent transcription. PMID- 11493685 TI - Crystal structure of the dimeric extracellular domain of human carbonic anhydrase XII, a bitopic membrane protein overexpressed in certain cancer tumor cells. AB - Overexpression of the zinc enzyme carbonic anhydrase (CA; EC ) XII is observed in certain human cancers. This bitopic membrane protein contains an N-terminal extracellular catalytic domain, a membrane-spanning alpha-helix, and a small intracellular C-terminal domain. We have determined the three-dimensional structure of the extracellular catalytic domain of human CA XII by x-ray crystallographic methods at 1.55-A resolution. The structure reveals a prototypical CA fold; however, two CA XII domains associate to form an isologous dimer, an observation that is confirmed by studies of the enzyme in solution. The identification of signature GXXXG and GXXXS motifs in the transmembrane sequence that facilitate helix-helix association is additionally consistent with dimeric architecture. The dimer interface is situated so that the active site clefts of each monomer are clearly exposed on one face of the dimer, and the C termini are located together on the opposite face of the dimer to facilitate membrane interaction. The amino acid composition of the active-site cleft closely resembles that of the other CA isozymes in the immediate vicinity of the catalytic zinc ion, but differs in the region of the nearby alpha-helical "130's segment." The structure of the CA XII-acetazolamide complex is also reported at 1.50-A resolution, and prospects for the design of CA XII-specific inhibitors of possible chemotherapeutic value are discussed. PMID- 11493686 TI - Mechanism of termination of DNA replication of Escherichia coli involves helicase contrahelicase interaction. AB - Using yeast forward and reverse two-hybrid analyses, we have discovered that the replication terminator protein Tus of Escherichia coli physically interacts with DnaB helicase in vivo. We have confirmed this protein-protein interaction in vitro. We show further that replication termination involves protein-protein interaction between Tus and DnaB at a critical region of Tus protein, called the L1 loop. Several mutations located in the L1 loop region not only reduced the protein-protein interaction but also eliminated or reduced the ability of the mutant forms of Tus to arrest DnaB at a Ter site. At least one mutation, E49K, significantly reduced Tus-DnaB interaction and almost completely eliminated the contrahelicase activity of Tus protein in vitro without significantly reducing the affinity of the mutant form of Tus for Ter DNA, in comparison with the wild type protein. The results, considered along with the crystal structure of Tus-Ter complex, not only elucidate further the mechanism of helicase arrest but also explain the molecular basis of polarity of replication fork arrest at Ter sites. PMID- 11493687 TI - Role of glutamate delta -2 receptors in activity-dependent competition between heterologous afferent fibers. AB - A principle that regulates detailed architecture in the brain is that active terminals have a competitive advantage over less active terminals in establishing synaptic connections. This principle is known to apply to fibers within a single neuronal population competing for a common target domain. Here we uncover an additional rule that applies when two neuronal populations compete for two contiguous territories. The cerebellar Purkinje cell dendrites have two different synaptic domains with spines innervated by two separate excitatory inputs, parallel fibers (PFs) and climbing fibers (CFs). Glutamate delta-2 receptors are normally present only on the PF spines where they are important for their innervation. After block of activity by tetrodotoxin, numerous new spines form in the CF domain and become innervated mainly by PFs; all spines, including those still innervated by the CFs, bear delta-2 receptors. Thus, in the absence of activity, PFs gain a competitive advantage over CFs. The entire dendritic arbor becomes a uniform territory with the molecular cues associated with the PFs. To access their proper territory and maintain synaptic contacts, CFs must be active and locally repress the cues of the competitor afferents. PMID- 11493688 TI - A metallothionein containing a zinc finger within a four-metal cluster protects a bacterium from zinc toxicity. AB - Zinc is essential for many cellular processes, including DNA synthesis, transcription, and translation, but excess can be toxic. A zinc-induced gene, smtA, is required for normal zinc-tolerance in the cyanobacterium Synechococcus PCC 7942. Here we report that the protein SmtA contains a cleft lined with Cys sulfur and His-imidazole ligands that binds four zinc ions in a Zn(4)Cys(9)His(2) cluster. The thiolate sulfurs of five Cys ligands provide bridges between the two ZnCys(4) and two ZnCys(3)His sites, giving two fused six-membered rings with distorted boat conformations. The inorganic core strongly resembles the Zn(4)Cys(11) cluster of mammalian metallothionein, despite different amino acid sequences, a different linear order of the ligands, and presence of histidine ligands. Also, SmtA contains elements of secondary structure not found in metallothioneins. One of the two Cys(4)-coordinated zinc ions in SmtA readily exchanges with exogenous metal ((111)Cd), whereas the other is inert. The thiolate sulfur ligands bound to zinc in this site are buried within the protein. Regions of beta-strand and alpha-helix surround the inert site to form a zinc finger resembling the zinc fingers in GATA and LIM-domain proteins. Eukaryotic zinc fingers interact specifically with other proteins or DNA and an analogous interaction can therefore be anticipated for prokaryotic zinc fingers. SmtA now provides structural proof for the existence of zinc fingers in prokaryotes, and sequences related to the zinc finger motif can be identified in several bacterial genomes. PMID- 11493689 TI - Bat predation on nocturnally migrating birds. AB - Bat predation on birds is a very rare phenomenon in nature. Most documented reports of bird-eating bats refer to tropical bats that occasionally capture resting birds. Millions of small birds concentrate and cross over the world's temperate regions during migration, mainly at night, but no nocturnal predators are known to benefit from this enormous food resource. An analysis of 14,000 fecal pellets of the greater noctule bat (Nyctalus lasiopterus) reveals that this species captures and eats large numbers of migrating passerines, making it the only bat species so far known that regularly preys on birds. The echolocation characteristics and wing morphology of this species strongly suggest that it captures birds in flight. PMID- 11493690 TI - Segregation of leading-edge and uropod components into specific lipid rafts during T cell polarization. AB - Redistribution of specialized molecules in migrating cells develops asymmetry between two opposite cell poles, the leading edge and the uropod. We show that acquisition of a motile phenotype in T lymphocytes results in the asymmetric redistribution of ganglioside GM3- and GM1-enriched raft domains to the leading edge and to the uropod, respectively. This segregation to each cell pole parallels the specific redistribution of membrane proteins associated to each raft subfraction. Our data suggest that raft partitioning is a major determinant for protein redistribution in polarized T cells, as ectopic expression of raft associated proteins results in their asymmetric redistribution, whereas non-raft partitioned mutants of these proteins are distributed homogeneously in the polarized cell membrane. Both acquisition of a migratory phenotype and SDF-1alpha induced chemotaxis are cholesterol depletion-sensitive. Finally, GM3 and GM1 raft redistribution requires an intact actin cytoskeleton, but is insensitive to microtubule disruption. We propose that membrane protein segregation not only between raft and nonraft domains but also between distinct raft subdomains may be an organizational principle that mediates redistribution of specialized molecules needed for T cell migration. PMID- 11493691 TI - Mapping the interactions between flavodoxin and its physiological partners flavodoxin reductase and cobalamin-dependent methionine synthase. AB - Flavodoxins are electron-transfer proteins that contain the prosthetic group flavin mononucleotide. In Escherichia coli, flavodoxin is reduced by the FAD containing protein NADPH:ferredoxin (flavodoxin) oxidoreductase; flavodoxins serve as electron donors in the reductive activation of anaerobic ribonucleotide reductase, biotin synthase, pyruvate formate lyase, and cobalamin-dependent methionine synthase. In addition, domains homologous to flavodoxin are components of the multidomain flavoproteins cytochrome P450 reductase, nitric oxide synthase, and methionine synthase reductase. Although three-dimensional structures are known for many of these proteins and domains, very little is known about the structural aspects of their interactions. We address this issue by using NMR chemical shift mapping to identify the surfaces on flavodoxin that bind flavodoxin reductase and methionine synthase. We find that these physiological partners bind to unique overlapping sites on flavodoxin, precluding the formation of ternary complexes. We infer that the flavodoxin-like domains of the cytochrome P450 reductase family form mutually exclusive complexes with their electron donating and -accepting partners, complexes that require conformational changes for interconversion. PMID- 11493692 TI - BRCA1 mediates ligand-independent transcriptional repression of the estrogen receptor. AB - Mutational inactivation of BRCA1 confers a cumulative lifetime risk of breast and ovarian cancers. However, the underlying basis for the tissue-restricted tumor suppressive properties of BRCA1 remains poorly defined. Here we show that BRCA1 mediates ligand-independent transcriptional repression of the estrogen receptor alpha (ERalpha), a principal determinant of the growth, differentiation, and normal functional status of breasts and ovaries. In Brca1-null mouse embryo fibroblasts and BRCA1-deficient human ovarian cancer cells, ERalpha exhibited ligand-independent transcriptional activity that was not observed in Brca1 proficient cells. Ectopic expression in Brca1-deficient cells of wild-type BRCA1, but not clinically validated BRCA1 missense mutants, restored ligand-independent repression of ERalpha in a manner dependent upon apparent histone deacetylase activity. In estrogen-dependent human breast cancer cells, chromatin immunoprecipitation analysis revealed the association of BRCA1 with ERalpha at endogenous estrogen-response elements before, but not after estrogen stimulation. Collectively, these results reveal BRCA1 to be a ligand-reversible barrier to transcriptional activation by unliganded promoter-bound ERalpha and suggest a possible mechanism by which functional inactivation of BRCA1 could promote tumorigenesis through inappropriate hormonal regulation of mammary and ovarian epithelial cell proliferation. PMID- 11493693 TI - DNA/DNA hybridization to microarrays reveals gene-specific differences between closely related microbial genomes. AB - DNA microarrays constructed with full length ORFs from Shewanella oneidensis, MR 1, were hybridized with genomic DNA from nine other Shewanella species and Escherichia coli K-12. This approach enabled visualization of relationships between organisms by comparing individual ORF hybridizations to 164 genes and is further amenable to high-density high-throughput analyses of complete microbial genomes. Conserved genes (arcA and ATP synthase) were identified among all species investigated. The mtr operon, which is involved in iron reduction, was poorly conserved among other known metal-reducing Shewanella species. Results were most informative for closely related organisms with small subunit rRNA sequence similarities greater than 93% and gyrB sequence similarities greater than 80%. At this level of relatedness, the similarity between hybridization profiles was strongly correlated with sequence divergence in the gyrB gene. Results revealed that two strains of S. oneidensis (MR-1 and DLM7) were nearly identical, with only 3% of the ORFs hybridizing poorly, in contrast to hybridizations with Shewanella putrefaciens, formerly considered to be the same species as MR-1, in which 63% of the ORFs hybridized poorly (log ratios below 0.75). Genomic hybridizations showed that genes in operons had consistent levels of hybridization across an operon in comparison to a randomly sampled data set, suggesting that similar applications will be informative for identification of horizontally acquired genes. The full value of microbial genomic hybridizations lies in providing the ability to understand and display specific differences between closely related organisms providing a window into understanding microheterogeneity, bacterial speciation, and taxonomic relationships. PMID- 11493694 TI - Mechanism of transfer of NO from extracellular S-nitrosothiols into the cytosol by cell-surface protein disulfide isomerase. AB - N-dansylhomocysteine (DnsHCys) is quenched on S-nitrosation. The product of this reaction, N-dansyl-S-nitrosohomocysteine, is a sensitive, direct fluorogenic substrate for the denitrosation activity of protein disulfide isomerase (PDI) with an apparent K(M) of 2 microM. S-nitroso-BSA (BSA-NO) competitively inhibited this reaction with an apparent K(I) of 1 microM. The oxidized form of DnsHCys, N,N-didansylhomocystine, rapidly accumulated in cells and was reduced to DnsHCys. The fluorescence of DnsHCys-preloaded human umbilical endothelial cells and hamster lung fibroblasts were monitored as a function of extracellular BSA-NO concentration via dynamic fluorescence microscopy. The observed quenching of the DnsHCys fluorescence was an indirect measure of cell surface PDI (csPDI) catalyzed denitrosation of extracellular S-nitrosothiols as decrease or increase in the csPDI levels in HT1080 fibrosarcoma cells correlated with the rate of quenching and the PDI inhibitors, 5,5'-dithio-bis-3-nitrobenzoate and 4-(N-(S glutathionylacetyl) amino)phenylarsenoxide inhibited quenching. The apparent K(M) values for denitrosation of BSA-NO by csPDI ranged from 12 microM to 30 microM. Depletion of membrane N(2)O(3) with the lipophylic antioxidant, vitamin E, inhibited csPDI-mediated quenching rates of DnsHCys fluorescence by approximately 70%. The K(M) for BSA-NO increased by approximately 3-fold and V(max) decreased by approximately 4-fold. These findings suggest that csPDI catalyzed NO released from extracellular S-nitrosothiols accumulates in the membrane where it reacts with O2 to produce N(2)O(3). Intracellular thiols may then be nitrosated by N2O3 at the membrane-cytosol interface. PMID- 11493696 TI - Gene expression profiling of clear cell renal cell carcinoma: gene identification and prognostic classification. AB - To better understand the molecular mechanisms that underlie the tumorigenesis and progression of clear cell renal cell carcinoma (ccRCC), we studied the gene expression profiles of 29 ccRCC tumors obtained from patients with diverse clinical outcomes by using 21,632 cDNA microarrays. We identified gene expression alterations that were both common to most of the ccRCC studied and unique to clinical subsets. There was a significant distinction in gene expression profile between patients with a relatively nonaggressive form of the disease [100% survival after 5 years with the majority (15/17 or 88%) having no clinical evidence of metastasis] versus patients with a relatively aggressive form of the disease (average survival time 25.4 months with a 0% 5-year survival rate). Approximately 40 genes most accurately make this distinction, some of which have previously been implicated in tumorigenesis and metastasis. To test the robustness and potential clinical usefulness of this molecular distinction, we simulated its use as a prognostic tool in the clinical setting. In 96% of the ccRCC cases tested, the prediction was compatible with the clinical outcome, exceeding the accuracy of prediction by staging. These results suggest that two molecularly distinct forms of ccRCC exist and that the integration of expression profile data with clinical parameters could serve to enhance the diagnosis and prognosis of ccRCC. Moreover, the identified genes provide insight into the molecular mechanisms of aggressive ccRCC and suggest intervention strategies. PMID- 11493697 TI - Nephrocystin interacts with Pyk2, p130(Cas), and tensin and triggers phosphorylation of Pyk2. AB - Juvenile nephronophthisis type 1 is caused by mutations of NPHP1, the gene encoding for nephrocystin. The function of nephrocystin is presently unknown, but the presence of a Src homology 3 domain and its recently described interaction with p130(Cas) suggest that nephrocystin is part of the focal adhesion signaling complex. We generated a nephrocystin-specific antiserum and analyzed the interaction of native nephrocystin with endogenous proteins. Immunoprecipitation of nephrocystin revealed that nephrocystin forms protein complexes with p130(Cas), proline-rich tyrosine kinase 2 (Pyk2), and tensin, indicating that these proteins participate in a common signaling pathway. Expression of nephrocystin resulted in phosphorylation of Pyk2 on tyrosine 402 as well as activation of downstream mitogen-activated protein kinases, such as ERK1 and ERK2. Our findings suggest that nephrocystin helps to recruit Pyk2 to cell matrix adhesions, thereby initiating phosphorylation of Pyk2 and Pyk2-dependent signaling. A lack of functional nephrocystin may compromise Pyk2 signaling in a subset of renal epithelial cells. PMID- 11493698 TI - Functional analysis of the LACERATA gene of Arabidopsis provides evidence for different roles of fatty acid omega -hydroxylation in development. AB - We describe lacerata (lcr) mutants of Arabidopsis, which display various developmental abnormalities, including postgenital organ fusions, and report cloning of the LCR gene by using the maize transposon Enhancer/Suppressor-mutator (En/Spm). The pleiotropic mutant phenotype could be rescued by genetic complementation of lcr mutants with the wild-type LCR gene. The LCR gene encodes a cytochrome P450 monooxygenase, CYP86A8, which catalyzes omega-hydroxylation of fatty acids ranging from C12 to C18:1, as demonstrated by expression of the gene in yeast. Although palmitic and oleic acids were efficient substrates for LCR, 9,10-epoxystearate was not metabolized. Taken together with previous studies, our findings indicate that LCR-dependent omega-hydroxylation of fatty acids could be implicated in the biosynthesis of cutin in the epidermis and in preventing postgenital organ fusions. Strikingly, the same pathway seems to control trichome differentiation, the establishment of apical dominance, and senescence in plants. PMID- 11493699 TI - Crossmodal binding of fear in voice and face. AB - In social environments, multiple sensory channels are simultaneously engaged in the service of communication. In this experiment, we were concerned with defining the neuronal mechanisms for a perceptual bias in processing simultaneously presented emotional voices and faces. Specifically, we were interested in how bimodal presentation of a fearful voice facilitates recognition of fearful facial expression. By using event-related functional MRI, that crossed sensory modality (visual or auditory) with emotional expression (fearful or happy), we show that perceptual facilitation during face fear processing is expressed through modulation of neuronal responses in the amygdala and the fusiform cortex. These data suggest that the amygdala is important for emotional crossmodal sensory convergence with the associated perceptual bias during fear processing, being mediated by task-related modulation of face-processing regions of fusiform cortex. PMID- 11493700 TI - p70S6 kinase signals cell survival as well as growth, inactivating the pro apoptotic molecule BAD. AB - Cytokines often deliver simultaneous, yet distinct, cell growth and cell survival signals. The 70-kDa ribosomal protein S6 kinase (p70S6K) is known to regulate cell growth by inducing protein synthesis components. We purified membrane-based p70S6K as a kinase responsible for site-specific phosphorylation of BAD, which inactivates this proapoptotic molecule. Rapamycin inhibited mitochondrial-based p70S6K, which prevented phosphorylation of Ser-136 on BAD and blocked cell survival induced by insulin-like growth factor 1 (IGF-1). Moreover, IGF-1-induced phosphorylation of BAD Ser-136 was abolished in p70S6K-deficient cells. Thus, p70S6K is itself a dual pathway kinase, signaling cell survival as well as growth through differential substrates which include mitochondrial BAD and the ribosomal subunit S6, respectively. PMID- 11493701 TI - Periodic perturbations in Shaker K+ channel gating kinetics by deletions in the S3-S4 linker. AB - Upon depolarization positive charges contained in the transmembrane segment S4 of voltage-dependent channels are displaced from the cytoplasmic to the external milieu. This charge movement leads to channel opening. In Shaker K+ channels four positively charged arginines in the S4 domain are transferred from the internal to the external side of the channel during activation. The distance traveled by the S4 segment during activation is unknown, but large movements should be constrained by the S3-S4 linker. Constructing deletion mutants, we show that the activation time constant and the midpoint of the voltage activation curve of the Shaker K+ channel macroscopic currents becomes a periodic function of the S3-S4 linker length for linkers shorter than 7 aa residues. The periodicity is that typical of alpha-helices. Moreover, a linker containing only 3 aa is enough to recover the wild-type phenotype. The deletion method revealed the importance of the S3-S4 linker in determining the channel gating kinetics and indicated that the alpha-helical nature of S4 extends toward its N terminus. These results support the notion that a small displacement of the S4 segment suffices to displace the four gating charges involved in channel opening. PMID- 11493702 TI - Identification of a family of Fc receptor homologs with preferential B cell expression. AB - Investigation of human genome sequences with a consensus sequence derived from receptors for the Fc region of Igs (FcR) led to the identification of a subfamily of five Ig superfamily members that we term the Fc receptor homologs (FcRHs). The closely linked FcRH genes are located in a chromosome 1q21 region in the midst of previously recognized FcR genes. This report focuses on the FcRH1, FcRH2, and FcRH3 members of this gene family. Their cDNAs encode type I transmembrane glycoproteins with 3-6 Ig-like extracellular domains and cytoplasmic domains containing consensus immunoreceptor tyrosine-based activating and/or inhibitory signaling motifs. The five FcRH genes are structurally related, and their protein products share 28-60% extracellular identity with each other. They also share 15 31% identity with their closest FcR relatives. The FcRH genes are expressed primarily, although not exclusively, by mature B lineage cells. Their conserved structural features, patterns of cellular expression, and the inhibitory and activating signaling potential of their transmembrane protein products suggest that the members of this FcRH multigene family may serve important regulatory roles in normal and neoplastic B cell development. PMID- 11493703 TI - Role of guanylate cyclase-activating proteins (GCAPs) in setting the flash sensitivity of rod photoreceptors. AB - The retina's photoreceptor cells adjust their sensitivity to allow photons to be transduced over a wide range of light intensities. One mechanism thought to participate in sensitivity adjustments is Ca(2+) regulation of guanylate cyclase (GC) by guanylate cyclase-activating proteins (GCAPs). We evaluated the contribution of GCAPs to sensitivity regulation in rods by disrupting their expression in transgenic mice. The GC activity from GCAPs-/- retinas showed no Ca(2+) dependence, indicating that Ca(2+) regulation of GCs had indeed been abolished. Flash responses from dark-adapted GCAPs-/- rods were larger and slower than responses from wild-type rods. In addition, the incremental flash sensitivity of GCAPs-/- rods failed to be maintained at wild-type levels in bright steady light. GCAP2 expressed in GCAPs-/- rods restored maximal light induced GC activity but did not restore normal flash response kinetics. We conclude that GCAPs strongly regulate GC activity in mouse rods, decreasing the flash sensitivity in darkness and increasing the incremental flash sensitivity in bright steady light, thereby extending the rod's operating range. PMID- 11493704 TI - Near-precise interchromosomal recombination and functional DNA topoisomerase II cleavage sites at MLL and AF-4 genomic breakpoints in treatment-related acute lymphoblastic leukemia with t(4;11) translocation. AB - We analyzed the der(11) and der(4) genomic breakpoint junctions of a t(4;11) in the leukemia of a patient previously administered etoposide and dactinomycin by molecular and biochemical approaches to gain insights about the translocation mechanism and the relevant drug exposure. The genomic breakpoint junctions were amplified by PCR. Cleavage of DNA substrates containing the normal homologues of the MLL and AF-4 translocation breakpoints was examined in vitro upon incubation with human DNA topoisomerase IIalpha and etoposide, etoposide catechol, etoposide quinone, or dactinomycin. The der(11) and der(4) genomic breakpoint junctions both involved MLL intron 6 and AF-4 intron 3. Recombination was precise at the sequence level except for the overall gain of a single templated nucleotide. The translocation breakpoints in MLL and AF-4 were DNA topoisomerase II cleavage sites. Etoposide and its metabolites, but not dactinomycin, enhanced cleavage at these sites. Assuming that DNA topoisomerase II was the mediator of the breakage, processing of the staggered nicks induced by DNA topoisomerase II, including exonucleolytic deletion and template-directed polymerization, would have been required before ligation of the ends to generate the observed genomic breakpoint junctions. These data are inconsistent with a translocation mechanism involving interchromosomal recombination by simple exchange of DNA topoisomerase II subunits and DNA-strand transfer; however, consistent with reciprocal DNA topoisomerase II cleavage events in MLL and AF-4 in which both breaks became stable, the DNA ends were processed and underwent ligation. Etoposide and/or its metabolites, but not dactinomycin, likely were the relevant exposures in this patient. PMID- 11493705 TI - DsbC activation by the N-terminal domain of DsbD. AB - The correct formation of disulfide bonds in the periplasm of Escherichia coli involves Dsb proteins, including two related periplasmic disulfide-bond isomerases, DsbC and DsbG. DsbD is a membrane protein required to maintain the functional oxidation state of DsbC and DsbG. In this work, purified proteins were used to investigate the interaction between DsbD and DsbC. A 131-residue N terminal fragment of DsbD (DsbDalpha) was expressed and purified and shown to form a functional folded domain. Gel filtration results indicate that DsbDalpha is monomeric. DsbDalpha was shown to interact directly with and to reduce the DsbC dimer, thus increasing the isomerase activity of DsbC. The DsbC-DsbDalpha complex was characterized, and formation of the complex was shown to require the N-terminal dimerization domain of DsbC. These results demonstrate that DsbD interacts directly with full-length DsbC and imply that no other periplasmic components are required to maintain DsbC in the functional reduced state. PMID- 11493706 TI - The Sec6/8 complex in mammalian cells: characterization of mammalian Sec3, subunit interactions, and expression of subunits in polarized cells. AB - The yeast exocyst complex (also called Sec6/8 complex in higher eukaryotes) is a multiprotein complex essential for targeting exocytic vesicles to specific docking sites on the plasma membrane. It is composed of eight proteins (Sec3, -5, -6, -8, -10, and -15, and Exo70 and -84), with molecular weights ranging from 70 to 144 kDa. Mammalian orthologues for seven of these proteins have been described and here we report the cloning and initial characterization of the remaining subunit, Sec3. Human Sec3 (hSec3) shares 17% sequence identity with yeast Sec3p, interacts in the two-hybrid system with other subunits of the complex (Sec5 and Sec8), and is expressed in almost all tissues tested. In yeast, Sec3p has been proposed to be a spatial landmark for polarized secretion (1), and its localization depends on its interaction with Rho1p (2). We demonstrate here that hSec3 lacks the potential Rho1-binding site and GFP-fusions of hSec3 are cytosolic. Green fluorescent protein (GFP)-fusions of nearly every subunit of the mammalian Sec6/8 complex were expressed in Madin-Darby canine kidney (MDCK) cells, but they failed to assemble into a complex with endogenous proteins and localized in the cytosol. Of the subunits tested, only GFP-Exo70 localized to lateral membrane sites of cell-cell contact when expressed in MDCK cells. Cells overexpressing GFP-Exo70 fail to form a tight monolayer, suggesting the Exo70 targeting interaction is critical for normal development of polarized epithelial cells. PMID- 11493707 TI - Hepatitis C virus replication is directly inhibited by IFN-alpha in a full-length binary expression system. AB - Hepatitis C virus (HCV) is a leading cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. The absence of culture systems permissive for HCV replication has presented a major bottleneck to antiviral development. We sought to recapitulate the early steps in the life cycle of HCV by means of DNA-based expression of viral genomic sequences. Here we report expression of replicating HCV RNA by using a, to our knowledge, novel binary expression system in which cells were transfected with a T7 polymerase-driven full-length HCV cDNA plasmid containing a cis-acting hepatitis Delta ribozyme to control 3' cleavage, and infected with vaccinia-T7 polymerase. HCV genomic and replicative strand synthesis, in addition to protein synthesis, was detectable and depended on full length HCV sequences. Moreover, the system was capable of generating HCV RNA quasispecies, consistent with the action of the low-fidelity HCV NS5B RNA polymerase. IFN-alpha, but not ribavirin, directly inhibited the viral replicative cycle in these cells, identifying the virus itself and not solely the immune system as a direct target of IFN action. The availability of a cell-based test for viral replication will facilitate screening of inhibitory compounds, analysis of IFN-resistance mechanisms, and analysis of virus-host cell interactions. PMID- 11493708 TI - Formation of a selenium-substituted rhodanese by reaction with selenite and glutathione: possible role of a protein perselenide in a selenium delivery system. AB - Selenophosphate is the active selenium-donor compound required by bacteria and mammals for the specific synthesis of Secys-tRNA, the precursor of selenocysteine in selenoenzymes. Although free selenide can be used in vitro for the synthesis of selenophosphate, the actual physiological selenium substrate has not been identified. Rhodanese (EC ) normally occurs as a persulfide of a critical cysteine residue and is believed to function as a sulfur-delivery protein. Also, it has been demonstrated that a selenium-substituted rhodanese (E-Se form) can exist in vitro. In this study, we have prepared and characterized an E-Se rhodanese. Persulfide-free bovine-liver rhodanese (E form) did not react with SeO(3)(2-) directly, but in the presence of reduced glutathione (GSH) and SeO(3)(2-) E-Se rhodanese was generated. These results indicate that the intermediates produced from the reaction of GSH with SeO(3)(2-) are required for the formation of a selenium-substituted rhodanese. E-Se rhodanese was stable in the presence of excess GSH at neutral pH at 37 degrees C. E-Se rhodanese could effectively replace the high concentrations of selenide normally used in the selenophosphate synthetase in vitro assay in which the selenium-dependent hydrolysis of ATP is measured. These results show that a selenium-bound rhodanese could be used as the selenium donor in the in vitro selenophosphate synthetase assay. PMID- 11493695 TI - Exploring the transcriptome of the malaria sporozoite stage. AB - Most studies of gene expression in Plasmodium have been concerned with asexual and/or sexual erythrocytic stages. Identification and cloning of genes expressed in the preerythrocytic stages lag far behind. We have constructed a high quality cDNA library of the Plasmodium sporozoite stage by using the rodent malaria parasite P. yoelii, an important model for malaria vaccine development. The technical obstacles associated with limited amounts of RNA material were overcome by PCR-amplifying the transcriptome before cloning. Contamination with mosquito RNA was negligible. Generation of 1,972 expressed sequence tags (EST) resulted in a total of 1,547 unique sequences, allowing insight into sporozoite gene expression. The circumsporozoite protein (CS) and the sporozoite surface protein 2 (SSP2) are well represented in the data set. A BLASTX search with all tags of the nonredundant protein database gave only 161 unique significant matches (P(N) < or = 10(-4)), whereas 1,386 of the unique sequences represented novel sporozoite-expressed genes. We identified ESTs for three proteins that may be involved in host cell invasion and documented their expression in sporozoites. These data should facilitate our understanding of the preerythrocytic Plasmodium life cycle stages and the development of preerythrocytic vaccines. PMID- 11493709 TI - Essential role of Drosophila Hdac1 in homeotic gene silencing. AB - Deacetylation of the N-terminal tails of core histones plays a crucial role in gene silencing. Rpd3 and Hda1 represent two major types of genes encoding trichostatin A-sensitive histone deacetylases. Although they have been widely found, their cellular and developmental roles remain to be elucidated in metazoa. We show that Drosophila Hdac1, an Rpd3-type gene, interacts cooperatively with Polycomb group repressors in silencing the homeotic genes that are essential for axial patterning of body segments. The biochemical copurification and cytological colocalization of HDAC1 and Polycomb group repressors strongly suggest that HDAC1 is a component of the silencing complex for chromatin modification on specific regulatory regions of homeotic genes. PMID- 11493710 TI - Neural agrin controls acetylcholine receptor stability in skeletal muscle fibers. AB - At mammalian neuromuscular junctions (NMJs), innervation induces and maintains the metabolic stability of acetylcholine receptors (AChRs). To explore whether neural agrin may cause similar receptor stabilization, we injected neural agrin cDNA of increasing transfection efficiencies into denervated adult rat soleus (SOL) muscles. As the efficiency increased, the amount of recombinant neural agrin expressed in the muscles also increased. This agrin aggregated AChRs on muscle fibers, whose half-life increased in a dose-dependent way from 1 to 10 days. Electrical muscle stimulation enhanced the stability of AChRs with short half-lives. Therefore, neural agrin can stabilize aggregated AChRs in a concentration- and activity-dependent way. However, there was no effect of stimulation on AChRs with a long half-life (10 days). Thus, at sufficiently high concentrations, neural agrin alone can stabilize AChRs to levels characteristic of innervated NMJs. PMID- 11493711 TI - Type I topoisomerase activity is required for proper chromosomal segregation in Escherichia coli. AB - Type I DNA topoisomerases are ubiquitous enzymes involved in many aspects of DNA metabolism. Escherichia coli possesses two type I topoisomerase activities, DNA topoisomerase I (Topo I) and III (Topo III). The gene encoding Topo III (topB) can be deleted without affecting cell viability. Cells possessing a deletion of the gene encoding Topo I (topA) are only viable in the presence of an additional compensatory mutation. In the presence of compensatory mutations, Topo I deletion strains grow normally; however, if Topo III activity is repressed in these cells, they filament extensively and possess an abnormal nucleoid structure. These defects can be suppressed by the deletion of the recA gene, suggesting that these enzymes may be involved in RecA-mediated recombination and may specifically resolve recombination intermediates before partitioning. PMID- 11493712 TI - Mitochondria play no roles in Mn(II)-induced apoptosis in HeLa cells. AB - Manganese(II) has been shown to exhibit catalase-like activity under physiological conditions. In the course of studies to test the antioxidant activity of Mn(II) on HeLa cells, it was observed at high concentrations (1-2 mM) that Mn(II) also induced apoptosis, as judged by changes in cell morphology, caspase-3 activation, cleavage of poly(ADP) ribose, and DNA condensation. However, in contrast to established mechanisms, the Mn(II)-induced apoptosis is associated with an increase rather than a decrease in mitochondrial inner membrane potential, as monitored by the fluorescent probe tetramethylrhodamine ethyl ester. Based on immunochemical analysis, Mn(II)-induced apoptosis does not lead to the release of cytochrome c into the cytosol. These and other measurements show that treatment with Mn(II) leads to enhancement of the mitochondrial "membrane mass," has no effect on mitochondrial volume, and does not affect the permeability transition pore. Together, these results support the view that Mn(II)-induced apoptosis occurs by a heretofore unrecognized mechanism. In addition, it was demonstrated that Mn(II) treatment leads to an increase in the production of reactive oxygen species (peroxides) and to the induction of the manganese superoxide dismutase and catalase activities but has no effect on the Cu,Zn-superoxide dismutase level. PMID- 11493713 TI - Heart regeneration in adult MRL mice. AB - The reaction of cardiac tissue to acute injury involves interacting cascades of cellular and molecular responses that encompass inflammation, hormonal signaling, extracellular matrix remodeling, and compensatory adaptation of myocytes. Myocardial regeneration is observed in amphibians, whereas scar formation characterizes cardiac ventricular wound healing in a variety of mammalian injury models. We have previously shown that the MRL mouse strain has an extraordinary capacity to heal surgical wounds, a complex trait that maps to at least seven genetic loci. Here, we extend these studies to cardiac wounds and demonstrate that a severe transmural, cryogenically induced infarction of the right ventricle heals extensively within 60 days, with the restoration of normal myocardium and function. Scarring is markedly reduced in MRL mice compared with C57BL/6 mice, consistent with both the reduced hydroxyproline levels seen after injury and an elevated cardiomyocyte mitotic index of 10-20% for the MRL compared with 1-3% for the C57BL/6. The myocardial response to injury observed in these mice resembles the regenerative process seen in amphibians. PMID- 11493715 TI - Gaucher's disease--an exemplary monogenic disorder. PMID- 11493714 TI - Arousal effect of orexin A depends on activation of the histaminergic system. AB - Orexin neurons are exclusively localized in the lateral hypothalamic area and project their fibers to the entire central nervous system, including the histaminergic tuberomammillary nucleus (TMN). Dysfunction of the orexin system results in the sleep disorder narcolepsy, but the role of orexin in physiological sleep-wake regulation and the mechanisms involved remain to be elucidated. Here we provide several lines of evidence that orexin A induces wakefulness by means of the TMN and histamine H(1) receptor (H1R). Perfusion of orexin A (5 and 25 pmol/min) for 1 hr into the TMN of rats through a microdialysis probe promptly increased wakefulness for 2 hr after starting the perfusion by 2.5- and 4-fold, respectively, concomitant with a reduction in rapid eye movement (REM) and non REM sleep. Microdialysis studies showed that application of orexin A to the TMN increased histamine release from both the medial preoptic area and the frontal cortex by approximately 2-fold over the baseline for 80 to 160 min in a dose dependent manner. Furthermore, infusion of orexin A (1.5 pmol/min) for 6 hr into the lateral ventricle of mice produced a significant increase in wakefulness during the 8 hr after starting infusion to the same level as the wakefulness observed during the active period in wild-type mice, but not at all in H1R gene knockout mice. These findings strongly indicate that the arousal effect of orexin A depends on the activation of histaminergic neurotransmission mediated by H1R. PMID- 11493716 TI - Refsum's disease. PMID- 11493718 TI - Increased oxysterols associated with iron accumulation in the brains and visceral organs of acaeruloplasminaemia patients. AB - Acaeruloplasminaemia is characterized by excessive neurovisceral iron accumulation due to mutation of the caeruloplasmin gene. Excess iron functions as a potent catalyst of biological oxidation, and increased iron concentration is associated with the products of lipid peroxidation in the serum and cerebrospinal fluid. We investigated whether the amount of iron accumulated paralleled lipid peroxidation levels in acaeruloplasminaemia tissues, examining brains and visceral organs of two affected patients at autopsy for iron and copper content, and oxysterols, including 7-hydroxycholesterol and 7-ketocholesterol, which are directly produced from cholesterol by active oxygen species. The amount of iron accumulated in various tissues was correlated with the levels of the oxysterols. These findings suggest that lipid peroxidation produced by the intracellular accumulation of iron is involved in the pathogenesis of acaeruloplasminaemia. PMID- 11493717 TI - Lung involvement and enzyme replacement therapy in Gaucher's disease. AB - Symptomatic lung involvement in Gaucher's disease is relatively rare, being restricted to patients with other severe manifestations. We describe our experience in eight of 411 patients in our referral clinic, who presented with prominent pulmonary signs or symptoms. There were four adults and four children; all have been successfully treated with enzyme replacement therapy. Routine means of monitoring pulmonary status including clinical assessment, chest X-ray, pulmonary function tests, and high-resolution CT (HRCT) were used. Enzyme treatment resulted in decreased hepatosplenomegaly, improved haematological parameters, and increased well-being; There was decreased clubbing and decreased dyspnoea in some of the patients, although on radiology, lung pathology had not normalized. All four children showed improved respiratory compliance, with significant improvement of the radiological findings in one and unchanged disease in the others. Two adults showed improvement in oxygen saturation but worsening of pulmonary hypertension. On chest X-ray, both had increased interstitial markings; one had gradual progression of pulmonary artery accentuation and fine interstitial stable pattern on HRCT. The other two adults had no change in lung function or on chest X-ray, but on HRCT there was apparent improvement in one patient. There is great heterogeneity in presentation and response to enzyme therapy in patients with Gaucher's disease and symptomatic lung involvement. Clinically, some benefited significantly from enzyme therapy, but in contrast to the dramatic reduction in organomegaly, there was no normalization in pulmonary function or lung architecture. PMID- 11493719 TI - Haematopoietic growth factor in antithyroid-drug-induced agranulocytosis. AB - Drug-induced agranulocytosis (DIA) is often caused by antithyroid drugs. We retrospectively studied the use of granulocyte colony-stimulating factor (G-CSF) therapy in antithyroid-DIA. Data for 20 patients (10 treated with G-CSF) with antithyroid-DIA (neutrophil count <0.5x10(9)/l) were extracted from a cohort study of DIA patients (n=110). G-CSF (300 microg/day subcutaneously) was used where the neutrophil count was <0.1x10(9)/l, or the patient was aged >70 years, or there were severe features of infection or underlying disease. Mean patient age was 62 years (range 34-87); sex ratio (M/F) was 0.05. Carbimazole (n=19) and benzylthiouracile (n=1) were the causative drugs, at mean doses of 30 mg/day (range 20-60) and 100 mg/day (range 50-150), respectively, for a mean of 37 days (range 31-90). Antithyroid drugs were prescribed for Graves' disease (n=8), thyrotoxicosis related to amiodarone intake (n=6) and multinodular goitre (n=6). Clinical features included isolated fever (n=7), pneumonia (n=5), septicaemia or septic shock (n=5) and acute tonsillitis (n=3). Mean neutrophil count was 0.07+/ 0.1x10(9)/l. No patient died. Mean durations of haematological recovery, antibiotic therapy and hospitalization were significantly reduced with G-CSF: 6.8+/-4 days vs. 11.6+/-5; 7.5+/-3.8 days vs. 12+/-4.5; and 7.3+/-4.8 days vs. 13+/-6.1, respectively (all p<0.05). G-CSF induced flu-like symptoms in 30% of patients, but reduced overall costs. PMID- 11493720 TI - Asthma admissions and thunderstorms: a study of pollen, fungal spores, rainfall, and ozone. AB - Asthma admissions have been reported to increase during thunderstorms. In some cases, this has been attributed to rises in pollen or fungal spore counts occurring alone or in combination with rainfall. We tested the hypothesis that thunderstorms in general are associated with asthma admissions, and investigated the possible roles of pollen, fungal spores, ozone, and other meteorological factors. We obtained data on multiple pollen and fungal spore counts, rainfall, temperature, ambient ozone concentrations, and asthma admissions for 32 dates when lightning strikes were recorded in the Cardiff/Newport area, and 64 matched dates in previous and subsequent years. Poisson regression models were used to investigate associations between admissions and proposed causative environmental factors. The number of asthma admissions was greater on days with thunderstorms than on control days (p<0.001). There were no associations or interactions between admissions and any pollen or fungal spore counts or rainfall. After adjusting for thunderstorms, there was an independent association between increasing ozone concentration, when temperature was included in the model, and increasing admissions (p=0.02). Asthma admissions are increased during thunderstorms. The effect is more marked in warmer weather, and is not explained by increases in grass pollen, total pollen or fungal spore counts, nor by an interaction between these and rainfall. There was an independent, positive association between ozone concentrations and asthma admissions. PMID- 11493721 TI - Preconception care and the risk of congenital anomalies in the offspring of women with diabetes mellitus: a meta-analysis. AB - Offspring of women with pregestational diabetes mellitus are at increased risk for congenital malformations, largely attributable to poor periconceptional glycaemic control. We assessed the effect of preconception care in reducing congenital malformations, in a meta-analysis of published studies of preconception care in women with diabetes mellitus. Articles were retrieved from Medline (1970 to June 2000) and Embase (1980 to June 2000), and data abstracted by two independent reviewers. The rates and relative risks (RR) for major and minor congenital malformations were pooled from all eligible studies using a random effects model, as were early first-trimester glycosylated haemoglobin values. In 14 cohort studies, major congenital malformations were assessed among 1192 offspring of mothers who had received preconception care, and 1459 offspring of women who had not. The pooled rate of major anomalies was lower among preconception care recipients (2.1%) than non-recipients (6.5%) (RR 0.36, 95%CI 0.22-0.59). In nine studies, the risk for major and minor anomalies was also lower among women who received preconception care (RR 0.32, 95%CI 0.17-0.59), as were the early first-trimester mean glycosylated haemoglobin values (pooled mean difference: 2.3%, 95%CI 2.1-2.4). Women who received preconception care were, on average, 1.8 years older than non-recipients, and fewer smoked (19.6% vs. 30.2%). Only one study described the routine use of periconception folic acid. Out patient preconception care probably reduces the risk of major congenital anomalies among the offspring of women with pregestational diabetes mellitus. Because many women with diabetes neither plan their pregnancy nor achieve adequate glycaemic control before conception, strategies are needed to improve access to these programs, and to maximize those interventions associated with improved pregnancy outcome, such as smoking cessation and folic acid use. PMID- 11493722 TI - Salicylic acid: a link between aspirin, diet and the prevention of colorectal cancer. AB - Aspirin was introduced into clinical practice more than 100 years ago. This unique drug belongs to a family of compounds called the salicylates, the simplest of which is salicylic acid, the principal metabolite of aspirin. Salicylic acid is responsible for the anti-inflammatory action of aspirin, and may cause the reduced risk of colorectal cancer observed in those who take aspirin. Yet salicylic acid and other salicylates occur naturally in fruits and plants, while diets rich in these are believed to reduce the risk of colorectal cancer. Serum salicylic acid concentrations are greater in vegetarians than non-vegetarians, and there is overlap between concentrations in vegetarians and those taking low dose aspirin. We propose that the cancer-preventive action of aspirin is due to its principal metabolite, salicylic acid, and that dietary salicylates can have the same effect. It is also possible that natural salicylates contribute to the other recognized benefits of a healthy diet. PMID- 11493723 TI - Bilateral frontal lobe lesions. PMID- 11493724 TI - Leucotomy: a qualified defence of 'then'. PMID- 11493725 TI - Transfusion medicine illustrated. What does the CD34+ cell look like? PMID- 11493726 TI - To test or not to test for syphilis: a global problem. PMID- 11493727 TI - Anemia-induced increase in the bleeding time: implications for treatment of nonsurgical blood loss. AB - BACKGROUND: Preoperative bleeding time (BT) does not correlate with postoperative bleeding in patients subjected to surgical procedures. A significant positive correlation has been reported between the BT 2 hours after cardiopulmonary bypass surgery and the nonsurgical blood loss during the first 4 hours after bypass surgery. This study was done to investigate the effect of Hct and platelet count on the BT measurement in normal, healthy men and women. STUDY DESIGN AND METHODS: To assess the relative effect of RBCs and platelets on the BT, 22 healthy male and 7 healthy female volunteers were subjected to the removal of 2 units of RBCs (360 mL), followed by the return of the platelet-rich plasma (PRP) from both units and the infusion of 1000 mL of 0.9-percent NaCl. Four of the men and all seven women received their RBCs 1 hour after their removal. Shed blood levels of thromboxane B(2) (TXB(2)), 6-keto prostaglandin F(1 alpha), and peripheral venous Hct were measured. BTs were measured in 15 men and 13 women before and after a plateletpheresis procedure to collect 3.6 x 10(11) platelets per unit. RESULTS: The 2-unit RBC apheresis procedure produced a 60-percent increase in the BT associated with a 15-percent reduction in the peripheral venous Hct and a 9 percent reduction in the platelet count. The plateletpheresis procedure produced a 32-percent decrease in the platelet count, no change in peripheral venous Hct, and no change in the BT. After the removal of 2 units of RBCs, the shed blood TXB(2) level decreased significantly. Reinfusion of 2 units of RBCs restored the BT and restored the TXB(2) level to the baseline levels. CONCLUSION: The acute reduction in Hct produced a reversible platelet dysfunction manifested by an increase in BT and a decrease in the shed blood TXB(2) level at the template BT site. Return of the RBCs restored both the BT and the shed blood TXB(2) level to normal. The platelet dysfunction observed with the reduction in Hct was due in part to a reduction in shed blood TXB(2) and other, unknown mechanisms. PMID- 11493728 TI - Platelet transfusion refractoriness associated with HPA-1a (Pl(A1)) alloantibody without coexistent HLA antibodies successfully treated with antigen-negative platelet transfusions. AB - BACKGROUND: Alloimmune-mediated refractoriness to platelet transfusion is most commonly due to antibody to HLA antigens in multiply transfused or multiparous patients. Published reports of poor transfusion response due to antibodies to platelet-specific antigens are rare and often confounded by the presence of coexistent antibodies against HLA antigens. CASE REPORT: A case is presented of a multiparous woman with acute myelogenous leukemia whose sole cause of transfusion refractoriness was antibody to platelet antigen HPA-1a. She responded dramatically to HPA-1a-negative platelet transfusion. CONCLUSION: This case provides strong serologic and clinical evidence that platelet transfusion refractoriness may result from antibodies to platelet-specific antigens. PMID- 11493729 TI - In vitro immunosuppressive activity of soluble HLA class I and Fas ligand molecules: do they play a role in autologous blood transfusion? AB - BACKGROUND: The immunomodulatory effects of allogeneic blood transfusion may contribute to a poor prognosis in patients with cancer who are undergoing surgery, and clinical trials have been carried out to investigate whether these patients would benefit from autologous blood donation. As the immunomodulatory effects of allogeneic blood transfusion have been related to soluble molecules released from residual WBCs during storage, the in vitro immunomodulatory activity of soluble molecules detected in supernatants from stored autologous blood was evaluated. STUDY DESIGN AND METHODS: Blood was donated by four healthy volunteers. Packed WBC-reduced RBCs were obtained and stored for 30 days, and supernatants were collected. FFP and serum were also obtained. The concentration of soluble molecules was determined by immunoenzymatic assays. The in vitro immunomodulatory activity of undiluted blood component supernatant was assessed by antigen-specific cytotoxic T-cell activity and mixed lymphocyte reactions in autologous combinations and by apoptosis induction in Fas+ cells. RESULTS: The concentrations of soluble Fas-ligand and HLA class I molecules were higher in packed RBCs than in WBC-reduced RBCs, FFP, and serum. Undiluted supernatants of packed RBCs strongly inhibited functional assays and induced apoptosis in Fas+ cells. The immunomodulatory effects were correlated with the amount of soluble Fas ligand and HLA class I molecules. CONCLUSION: The results of the present study are comparable with those already reported in allogeneic blood components, and they indicate that undiluted supernatants of autologous blood components may exert immunosuppressive effects in vitro. PMID- 11493730 TI - Prestorage universal WBC reduction of RBC units does not affect the incidence of transfusion reactions. AB - BACKGROUND: Febrile nonhemolytic transfusion reaction (FNHTR) has been identified as a pivotal reason for prestorage universal WBC reduction. A regional blood center implemented universal prestorage WBC reduction for RBCs on January 1, 2000. Whether prestorage universal WBC reduction of RBC units will affect FNHTR is not known. STUDY DESIGN AND METHODS: All reports of RBC transfusion reactions at Barnes-Jewish Hospital submitted for evaluation to the blood bank, before and after the implementation of WBC reduction of RBCs, were retrospectively evaluated. RESULTS: For the 36,303 allogeneic RBC transfusions administered in 1999, 85 reactions (0.23%) were reported. These reactions were classified as FNHTR in 43 cases, allergic in 13, delayed hemolytic in 19, and miscellaneous in 10. For the 31,543 non-WBC-reduced RBC transfusions performed in 1999, 78 reactions (0.25%) were reported. These reactions were classified as FNHTR in 39 cases, allergic in 13, delayed hemolytic in 19, and miscellaneous in 7. In the first half of 2000, 32 reactions (0.20%) were reported for 16,093 prestorage WBC reduced RBC transfusions (p = 0.41). There were 13 FNHTRs and 10 allergic, 7 delayed hemolytic, and 2 miscellaneous reactions. The use of prestorage WBC reduced RBCs did not significantly affect the rate of reactions classified as allergic (0.04% in 1999; 0.06% in 2000; p = 0.43) or as FNHTR (0.12% in 1999; 0.08% in 2000; p = 0.33). For all patients, universal WBC reduction in 2000 did not reduce the rate of FNHTR from the rate seen with selective bedside WBC reduction, the practice used in 1999 (0.12% in 1999; 0.08% in 2000; p = 0.36). CONCLUSION: No significant difference was found in the incidence of transfusion reactions in patients receiving prestorage WBC-reduced RBCs and non-WBC-reduced RBCs. In addition, no difference was found in transfusion reaction rates when periods of prestorage universal WBC reduction were compared to those of selective WBC reduction. PMID- 11493731 TI - Quantitative and genotypic analysis of TT virus infection in Chinese blood donors. AB - BACKGROUND: The TT virus (TTV) is a member of a newly described family of human viruses related to the C ircoviridae viruses. Its association with specific diseases has not been established, and screening of blood donors has not been implemented. To date, 16 genotypes have been identified. STUDY DESIGN AND METHODS: Sera from 471 healthy blood donors (aged 11-58 years) were randomly selected and tested for TTV by the use of two sets of primers: NG59d/NG61d/NG63d primers and T801/T935 primers. Quantitative competitive PCR (QC-PCR) was developed to measure the TTV DNA concentration among the blood donors. Sequencing of a part of the genome was performed to identify the various genotypes. Several samples showed a mixed genotype infection. RESULTS: TTV was detected in 251 (53.3%) of 471 healthy Hong Kong blood donors by the use of NG59d/NG61d/NG63d primers. The prevalence of the virus increased steadily with age (p = 0.03). TTV DNA was detected in 90 percent (90 of a randomly selected 100) of samples by the use of T801/T935 primers. TTV DNA concentration was also measured by QC-PCR in the blood donors who were positive for TTV DNA in the first round of the heminested PCR. TTV titers ranged from 4.8 x 10(2) copies per mL to 6 x 10(4) copies per mL, with a median value of 1.2 x 10(4) copies per mL. Sequencing and phylogenetic analysis of a 223-bp fragment from open reading frame 1 showed three main genotypes (G1 [60.7%], G2 [24.3%], and G3 [14%]) and a new genotype 17 (G17), with the latter bearing 60-percent nucleotide homology with other genotypes deposited at GenBank. In addition, a new TTV subtype, G2f, was found. CONCLUSION: The prevalence of TTV is high in healthy Chinese blood donors. Three main genotypes (G1, G2, and G3) were detected. In addition, a new TTV genotype, tentatively designated as G17, and a new subtype, G2f, were identified. PMID- 11493732 TI - Inverse relationship between patient peripheral blood CD34+ cell counts and collection efficiency for CD34+ cells in two automated leukapheresis systems. AB - BACKGROUND: The purpose of this study was to analyze the CD34 cell collection efficiency (CE) of automated leukapheresis protocols of two blood cell separators (Spectra, COBE [AutoPBSC protocol] and AS104, Fresenius [PBSC-Lym, protocol]) for peripheral blood progenitor cell (PBPC) harvest in patients with malignant diseases. STUDY DESIGN AND METHODS: PBPCs were collected by the Spectra AutoPBSC protocol in 95 patients (123 collections) and the AS104 PBSC-Lym protocol in 87 patients (115 harvests). Patients underwent a median of one (range, 1-4) conventional-volume apheresis procedure of 10.8 L (9.0-13.9) to obtain a target cell dose of > or =2.5 x 10(6) CD34+ cells per kg. RESULTS: The median overall CD34 CE was significantly better on the AS104 than on the Spectra: 55.8 percent versus 42.4 percent (p = 0.000). This was also true below (59.2% vs. 50.1%; p = 0.022) and above (51.2% vs. 41.3%; p = 0.001) the preleukapheresis threshold of 40 CD34+ cells per microL needed to collect a single-apheresis autograft. However, at > or =40 circulating CD34+ cells per microL, both cell separators achieved the target of > or =2.5 x 10(6) CD34+ cells per kg. The CD34 CE dropped significantly, from 59.2 percent at <40 cells per microL to 51.2 percent at > or =40 cells per microL on the AS104 (p = 0.017) and from 50.1 percent to 41.3 percent on the Spectra (p = 0.033). CONCLUSION: Whereas the CD34 CE was significantly different with the AS104 and the Spectra, the CD34 CE of both machines correlated inversely with peripheral blood CD34+ cell counts, showing a significant decline with increasing numbers of circulating CD34+ cells. Nevertheless, at > or 40 preapheresis CD34+ cells per microL, sufficient hematopoietic autografts of > or =2.5 x 10(6) CD34+ cells per kg were harvested by a single conventional-volume (11 L) leukapheresis on both cell separators. PMID- 11493733 TI - Repopulating activities of human cord blood cells separated by a stem cell collection filter in NOD/SCID mice: a comparative study of filter method and HES method. AB - BACKGROUND: Volume reduction and removal of RBCs are essential for cost-efficient cord blood (CB) banking. It has previously been shown that a newly developed device, a stem cell-collection filter (SCCF), can reduce the CB volume and remove RBCs efficiently, giving high recovery rates for CD34+ cells, colony-forming cells, and long-term culture-initiating cells with short operation time. The aim of this study was to compare the quality of CB cells separated by SCCF and HES by analyzing repopulation in NOD/SCID mice. STUDY DESIGN AND METHODS: A total of 1 x 10(6) or 5 x 10(6) nucleated cells derived from SCCF- or HES-separated, cryopreserved, thawed, and washed CB were transplanted into NOD/SCID mice. Eight weeks after transplantation, bone marrow cells of the recipient mice were examined by flow cytometry and hematopoietic progenitor assay for the engraftment of human cells. RESULTS: Mice given human CB cells, separated by SCCF, showed degrees of engraftment similar to those in mice given HES-separated CB cells. There was no significant difference in the lymphohematopoietic reconstitution pattern in the two groups of mice. CONCLUSION: SCCF processing does not appear to reduce the number of repopulating cells in NOD/SCID mice or alter the number of HPCs. It is now shown that these cells can be captured by SCCF and removed, and that they will engraft. PMID- 11493734 TI - Causality assessment of suspected virus transmission by human plasma products. AB - BACKGROUND: Causality assessment of reports on suspected virus transmission is crucial for early detection of infectious plasma products. Commonly used algorithms, such as the WHO criteria, do not meet the specific requirements for causality assessment of suspected virus transmission. STUDY DESIGN AND METHODS: A special algorithm, based on nucleic acid amplification and gene sequencing technology, effectiveness of validated virus-inactivation methods, empirical data concerning the safety record of the product, and information on batch-related infection clusters, was developed. The algorithm is focused on laboratory test results or otherwise standardized data, with few clinical data being required. To facilitate practical application, the algorithm has been converted into a graphical decision tree. RESULTS: The feasibility of the algorithm is shown by causality assessment of sample cases. Three cases are presented with the details of each case used in the 12-question checklist. The answers provided by the checklist led to the causality classification. CONCLUSION: The algorithm is a tool for evaluating reports of suspected virus transmission in a standardized manner. It thus has the potential to improve early signal detection in pharmacovigilance of plasma products by confirmation or exclusion of suspected infectivity in most cases. PMID- 11493735 TI - WBC reduction in RBC concentrates by prestorage filtration: multicenter experience. AB - BACKGROUND: As universal leukocyte (WBC) reduction (ULR) is being considered as a new standard, few data are available on the performance of WBC-reduction filtration in routine practice. The performance of WBC-reduction in RBCs, using varied filtration practices, in meeting the current FDA requirement (<5 x 10(6)), Council of Europe (EC) recommendation, the proposed FDA requirement (<1 x 10(6)), and a more stringent proposal (<5 x 10(5)) for residual WBCs per RBC unit was assessed and compared. STUDY DESIGN AND METHODS: Participating facilities were the 11 sites of the Viral Activation Transfusion Study (VATS), a prospective study of the impact of transfusion with and without WBC-reduction on survival and HIV viral load in HIV-1-infected patients. Patients randomly assigned to undergo WBC reduction were required to receive RBCs < or =14 days old that had undergone prestorage (within 72 hours of collection) WBC-reduction filtration by a method devised to achieve a postfiltration WBC count of <5 x 10(6). Residual WBC quantitation was performed by PCR in the central VATS laboratory by using frozen WBC-reduced RBC samples obtained at issue for transfusion. RESULTS: A total of 1869 WBC-reduced RBC units were studied. Filtration practices varied within and between sites. There were significant differences in mean residual WBC counts at the 11 sites (p<0.001). Among the WBC-reduced RBC units, 0.8 percent exceeded 5 x 10(6) WBCs per unit, 8.3 percent exceeded 1 x 10(6) WBCs per unit, and 14.3 percent exceeded 5 x 10(5) WBCs per unit. CONCLUSION: Residual WBCs in WBC reduced RBC units vary within and between sites. WBC reduction was successful, in that over 99 percent and 91 percent of VATS WBC-reduced RBC units met US and EC thresholds, respectively. However, the small but measurable failure rate indicates that not every unit will meet these guidelines. PMID- 11493736 TI - Administration of G--CSF plus dexamethasone produces greater granulocyte concentrate yields while causing no more donor toxicity than G--CSF alone. AB - BACKGROUND: G-CSF with or without dexamethasone is becoming the standard agent for mobilizing granulocytes for transfusion. The purpose of this study was to determine if the toxicities of G--CSF with or without dexamethasone are offset by greater collection yields and to define the minimum interval that should separate sequential collections. STUDY DESIGN AND METHODS: Twenty donors were studied on three occasions. They were given either dexamethasone (8 mg, by mouth) plus a placebo injection, G--CSF (5 microg/kg, given subcutaneously) plus placebo capsules, or G--CSF plus dexamethasone. Granulocytes were collected by apheresis. A donor symptom survey was administered, and cell counts and blood chemistries were assessed before collection and 1, 2, 7, 14, 21, 28, and 35 days after collection. RESULTS: More granulocytes were collected when G--CSF was given than when dexamethasone was given (41.1 +/- 20.4 x 10(9) vs. 21.0 +/- 10.0 x 10(9); p<0.001), but the use of G--CSF plus dexamethasone produced the greatest yields (67.1 +/- 22.0 x 10(9); p<0.002). When the donors were given dexamethasone alone, 58 percent experienced at least one symptom, compared to 85 percent of those given G--CSF and 75 percent of those given G--CSF plus dexamethasone. In all three regimens, platelet counts fell 19 percent to 24 percent after collection and remained below baseline for 7 to 14 days. Granulocyte counts returned to baseline within 3 to 7 days, but, in all three regimens, a mild granulocytopenia occurred 21 days after collection. With each of the regimens, blood chemistries changed, but the changes were mild and most returned to baseline within 7 days; however, changes in albumin, bilirubin, and AST persisted until 28 days after collection. CONCLUSION: These results support the use of G--CSF plus dexamethasone in granulocyte donors. G--CSF plus dexamethasone resulted in greater granulocyte yields than either agent alone and was associated with donor symptoms and changes in blood cell counts and chemistries similar to those seen with G--CSF alone or dexamethasone alone. Granulocytes can be safely collected a second time after a 7-day interval; however, for regular donors, it may be best to separate collections by 4 weeks. PMID- 11493737 TI - The role of electrolytes and pH in RBC ASs. AB - BACKGROUND: Experimental additive solutions (EASs) containing saline, adenine, glucose, mannitol and disodium phosphate can support RBCs for 9 or 10 weeks if used in 200- or 300-mL volumes. The effects of variations in the electrolyte composition and volume of EASs were explored. STUDY DESIGN AND METHODS: In three four-arm studies, 24 RBC units were pooled in groups of 4 and realiquoted as test units to ensure that all donors were equally represented in each study arm. In Study 1, units were stored for 11 weeks in EAS containing 0, 10, 20, or 30 mmol per L of sodium bicarbonate. In Study 2, units were stored for 9 weeks in EAS containing 26, 50, 100, or 150 mmol per L of sodium chloride. In Study 3, units were stored in 100 or 200 mL of AS-3 or EAS-61. RBC ATP concentrations and hemolysis were measured weekly. RESULTS: Increasing the sodium bicarbonate content of EASs increased the pH throughout storage and increased RBC ATP concentrations in the later phases of storage, but it had no effect on hemolysis. Increased sodium chloride content of EASs led to lower RBC ATP concentrations and increased hemolysis. In EAS-61, RBC ATP concentrations were increased throughout storage, and hemolysis was lower than that of RBCs stored in AS-3. CONCLUSION: RBC ATP synthesis is highly dependent on the pH of the AS. Hemolysis is affected by the salt content and volume of the AS. PMID- 11493738 TI - A D(V)-like phenotype is obliterated by A226P in the partial D DBS. AB - BACKGROUND: In D category V types, the RHD exon 5 or parts thereof are replaced by the corresponding RHCE DNA segments. In D category V types I and II, the amino acid at position 226 is alanine, which is typical of the prevalent RHD allele and is observed in all RHCE alleles encoding the antigen e. A proline at position 226 in RHCE encodes the antigen E. STUDY DESIGN AND METHODS: A blood sample of ccDEe phenotype was referred as suspected D category VI. The RHD nucleotide sequence and the D epitope pattern were determined. RESULTS: A new partial D, DBS, encoded by an RHD-RHcE(5)-RHD hybrid allele, was found. Although it differed from D(Va) type II by an A226P substitution only, it lacked epitopes epD4, epD12, epD17, epD18, and epD22 that were present in D(Va). The 5' breakpoint region was located between the deletion in RHD intron 4 and the first polymorphic nucleotide of DBS exon 5. CONCLUSION: The phenotypes of RHD alleles with gene conversions limited to exon 5 depended critically on the amino acid at position 226. If alanine was present at this position, gene conversions involving E233Q led to a D(Va)-like phenotype. If proline was present, many additional epitopes were lost, and the phenotype became reminiscent of DFR. The 5' breakpoint region is shared by 10 alleles and may represent the most active "hot spot" for gene conversions known in RH. PMID- 11493739 TI - Labeling D+ RBCs for flow cytometric quantification of fetomaternal hemorrhage after the RBCs have been coated with anti-D. AB - BACKGROUND: D- patients may receive Rh immunoglobulin (RhIg) before blood samples are taken for estimation of the volume of fetomaternal hemorrhage (FMH) by flow cytometry. Anti-D bound to the fetal D+ cells may then block the binding of conjugated D MoAb. This may reduce the fluorescence of the D+ cells, which would lead to ambiguity over setting the positions of the markers on histograms and may result in erroneous values of FMH. STUDY DESIGN AND METHODS: Labeling methods were compared by using FITC-BRAD 3 (anti-D) and/or FITC-anti-IgG (Fab fragment) with mixtures of D+ (R1r) and D- (rr) cells when the D+ cells had first been coated with various amounts (0 molecules/cell and 600-13,000 molecules/cell) of anti-D (RhIg). Variables examined were antibody concentrations, the order and times of incubation with the antibodies, and the effect of washing between the uptake of the antibodies used. RESULTS: In all cases, D+ cells were strongly labeled after incubation with 50 microL of FITC-BRAD-3 and then after washing with 50 microL of FITC-anti-human IgG, with both incubations being for 30 minutes at 37 degrees C. With this double-staining procedure, the fluorescence of D+ cells was found to be similar regardless of how much anti-D (RhIg) was previously bound and greater than that with FITC-BRAD-3 alone, giving an enhanced signal-to noise ratio. CONCLUSION: As the testing laboratory may not know if the patient has received prophylactic RhIg, this labeling method would be suitable for all samples. PMID- 11493741 TI - Syphilis, a disease of direct transfusion. PMID- 11493740 TI - Effects of colloidal resuscitation fluids on the neutrophil respiratory burst. AB - BACKGROUND: Experimental studies have revealed that gelatin and HES produce increased neutrophil respiratory burst activity. It was investigated whether 3 percent gelatin (MW 35,000) and three types of 6-percent HES (MW 70,000; degree of substitution, 0.5; 200,000/0.5; 450,000/0.7) preparations can influence superoxide anion production during respiratory burst under clinical conditions. STUDY DESIGN AND METHODS: Blood samples were obtained from 40 patients before and 1 hour after the infusion, before anesthesia and surgical treatment. After stimulation with bacteria (Escherichia coli), the respiratory burst was measured by oxidation of nonfluorescent dihydrorhodamine 123 to the fluorescent rhodamine 123 by the use of flow cytometry. RESULTS: Respiratory burst activity decreased significantly (p = 0.004) from the baseline (60.0 +/- 6.5%) to 1 hour after the administration of the low-molecular-weight HES preparation (55.0 +/- 6.8%). No significant differences in respiratory burst activity could be found after the administration of gelatin or medium-molecular-weight or high- molecular-weight HES solution. CONCLUSION: The investigated administration of gelatin and medium- and high-molecular-weight HES preparations did not influence respiratory burst activity under clinical conditions. However, the neutrophil respiratory burst was impaired after the administration of low-molecular-weight HES. Neutrophil respiratory burst activity may vary according to the type of colloidal plasma substitutes administered. PMID- 11493742 TI - The quality of RBCs stored in CPD/ADSOL. PMID- 11493743 TI - Erythrophagocytosis on the peripheral blood smear and paroxysmal cold hemoglobinuria. PMID- 11493744 TI - Detection of HBV DNA-positive HBsAg-negative platelet concentrate by routine PCR screening. PMID- 11493746 TI - Editorial. PMID- 11493745 TI - A fatal case of transfusion-transmitted Staphylococcus epidermidis sepsis. PMID- 11493747 TI - The Integration of Genomics into Public Health Research, Policy and Practice in the United States. AB - Objectives: To examine the opportunities for and responsibilities of the public health community in bridging the gap between gene discovery and the application of genetic information to improve health and prevent disease. Methods: We developed genetics-related definitions for the core functions and essential services of public health. We combined these definitions with a visual model to create one possible 'blueprint' for integrating genomics into public health activities. Results: The proposed blueprint and accompanying examples illustrate the important role for genomics throughout public health research, policy and practice. Further refinement and implementation of this blueprint represents an ambitious public health leadership agenda. Conclusions: Opportunities for immediate action include strategic planning for the integration of genomics across programs, developing genomics competencies among health professionals, enhancing surveillance and epidemiologic capacity to aid evidence-based policy making, building partnerships and seeking input from stakeholders and incorporating information about genomics into health communications. Copyright 2001 S. Karger AG, Basel PMID- 11493748 TI - Current Challenges to Appropriate Clinical Use of New Genetic Knowledge in Different Countries. AB - Objective: To describe the challenges facing countries all over the world regarding the appropriate clinical use of genetics in their health care systems. Methods and Results: Aspects of the economic and social contexts in different countries which are of particular relevance to shaping the existing challenges are outlined. Issues which are relevant (but of different prominence) in all countries in providing genetic services are discussed. Conclusions: The challenges facing the provision of appropriate genetic services differ markedly in four major groups of countries. These challenges range from controlling inappropriate commercialization and the overuse of genetic approaches to putting in place even minimal basic community genetics services in countries where the infant mortality rate has fallen to a range where genetic and congenital disorders contribute substantially to ongoing handicap and early mortality. Copyright 2001 S. Karger AG, Basel PMID- 11493749 TI - Public attitudes regarding the donation and storage of blood specimens for genetic research. AB - OBJECTIVE: As sequencing of the human genome is completed, there is a need for population-based research to assess frequencies of genetic variants and their associations with human diseases. The authors therefore assessed the current climate regarding the donation and storage of blood for genetic research. METHODS: Data from the American Healthstyles Survey fo health attitudes and behavior were examined. In the 1998 survey, four questions regarding blood donation and storage for genetic research were posed to the participants. RESULTS: Of 3,130 participants, 2,621 (84%) completed these questions. Of the respondents, 42% were in favor of both blood donation and long-term storage for genetic research, 37% were in favor of either blood donation or storage but not both and 21% were not willing to donate blood or have it stored for genetic research under any circumstances. Loglinear analysis demonstrated that the characteristics of respondents who favored blood donation and long-term storage for genetic research were attitudinal; specifically, those believing that genetic research will prevent disease [odds ratio (OR) 2.9; p<0.001]; those believing in genetic determinism (OR 1.5; p=0.004) and those agreeing they would participate in government research (OR 2.9; p<0.001). The model also demonstrated that characteristics indirectly associated with attitudes toward blood donation/storage for genetic research were demographic and included higher education, white race, living in the Mountain/Pacific or mid-Atlantic regions of the United States and positive family history of a genetic disorder (p<0.05). CONCLUSION: Understanding the various factors contributing to knowledge, attitudes and behavior regarding the donation and storage of blood specimens for genetic research will contribute to future actions in communication genetic research goals to the public and recruitment for population-based genetic studies. PMID- 11493750 TI - Finnish physicians' interest in genetic screening. AB - OBJECTIVES: To report on Finnish physicians' opinions on the possible future use of genetic screening and their interest in knowing their own risk for diseases. METHODS: A questionnaire survey mailed to gynaecologists, paediatricians, general practitioners and clinical geneticists. Both leading physicians and a sample of other practitioners were included (response rate 74%, n=571). RESULTS: Physicians preferred genetic testing of people at risk over population-wide screening. Breast and colon cancer, familial hypercholesterolaemia and juvenile diabetes were diseases for which screening was most often considered useful. The most popular target group was the whole population. Opinions on the stability of genetic screening were not uniform, and physicians did not take a stand on genetic screening as a whole but differentiated by disease and target group. Those wanting to know their own risk for diseases supported screening procedures in general more than did other physicians. CONCLUSIONS: Physicians were not enthusiastic about genetic screening. Nevertheless, testing of individuals with a family history of certain diseases is likely to become more widespread. PMID- 11493751 TI - Disease knowledge and attitudes toward predictive testing and prenatal diagnosis in families with Machado-Joseph disease from the Azores Islands (Portugal). AB - OBJECTIVE: Machado-Joseph disease (MJD) reaches its highest prevalence world-wide in the Azores, thus constituting a public health problem in these islands. The aim of the study was thus to (1) determine the level of knowledge about the disease; (2) estimate the expected level of request for predictive testing, and (3) analyse the intentions of at-risk individuals concerning their reproductive decisions. METHODS: A questionnaire on these points was distributed to 42 affected and 36 at-risk individuals. RESULTS: As expected, the educational level of the respondents was significantly associated with the level of knowledge about the disease. The survey indicated that 83.3% of the at-risk individuals would make use of predictive test and that 77.8% would make use of prenatal diagnosis. Of the latter, 36.1% would terminate pregnancy if confronted with a positive result for the fetus. CONCLUSIONS: The level of knowledge about MJD in the Azorean families is considered to be fair. Although the actual behavior can prove to be different from the intentions put forward by at-risk individuals based solely on the results of this study we can estimate that the request for a predictive test would be quite high. The intentions expressed by at-risk individuals seem to indicate that the prenatal diagnosis will have an effect on their reproductive decisions. Results obtained certify the importance of implementing genetic testing for MJD in the Azores. PMID- 11493752 TI - Triple Test Screening for Down Syndrome: Looking Back on a False-Positive Result and Having or Not Having a Triple Test in Subsequent Pregnancies. AB - Objectives: It was the main aim of the present retrospective study carried out in Flanders to evaluate how women with a false-positive triple test result look back on their experience and decision making and how many of them make use of the triple test in subsequent pregnancies. Methods: All 508 women tested in the Centre for Human Genetics in Leuven in 1995 who had a positive triple test result followed by a normal amniocentesis outcome were invited to participate in a mailed questionnaire study with open and multiple choice questions in 1998. The response rate was 68%. Results: The answers to the multiple choice question assessing how they look back on their initial expectations regarding the exact meaning of the triple test revealed that less than one half reported that it concerned the identification of 'a higher risk of carrying a child with Down syndrome (DS)'. Reporting correct initial expectations was significantly associated with a higher education level. The same holds for indecisiveness regarding pregnancy termination should the amniocentesis have detected a fetus with DS. As expected, a large majority of the women reported a high level of distress or worry after the communication of the positive triple test result. Overall the findings show that retrospectively most women had the feeling that the decision to have amniocentesis was their own decision rather than a professional's. Of the subgroup with one or more subsequent pregnancies 70% had another triple test. Conclusions: The overall results of this study clearly reveal a need for a systematic approach aimed at better informing and counselling pregnant women about the implications and limitations of the triple test. Notwithstanding the reported high level of distress caused by a positive triple test result, a large majority of the women with subsequent pregnancies had another triple test; they represent a clearly higher percentage than in another recent study. Copyright 2001 S. Karger AG, Basel PMID- 11493753 TI - The Tyr978X BRCA1 Mutation in Non-Ashkenazi Jews: Occurrence in High-Risk Families, General Population and Unselected Ovarian Cancer Patients. AB - Background: In Jewish individuals of Ashkenazi (East European) decent, three predominant mutations, 185 delAG and 5382insC (BRCA1) and 6174delT (BRCA2), seem to account for a substantial portion of germline mutations in high-risk breast/ovarian cancer families. Among non-Ashkenazi Jews, the 185delAG and the Tyr978X mutations, as well as several 'private' mutations have been reported within the BRCA1 gene. Objective: Assessing the occurrence rate of the Tyr978X BRCA1 germline mutation in Jewish non-Ashkenazi individuals: high-risk familial cases, unselected ovarian cancer patients and the general average risk Jewish Iraqi population. In addition, finding proof that this is a founder mutation. Methods: PCR amplification of the relevant fragment of the BRCA1 gene from constitutional DNA followed by restriction enzyme digest that differentiates the wild type from the mutant allele. In addition, BRCA1-linked markers were used for haplotype analysis. Results: The Tyr978X BRCA1 mutation was detected in 3/289 (1%) of the average-risk Jewish Iraqi population, in 7/408 (1.7%) high-risk Jewish non-Ashkenazi individuals (representing 332 unrelated families) and in 1/81 (1.2%) of unselected Jewish non-Ashkenazi ovarian cancer patients. Allelotyping using BRCA1-linked markers revealed an identical allelic pattern in all mutation carriers with the intragenic markers. Conclusions: Our findings suggest that this mutation is prevalent in Iraqi Jews, represents a founder mutation, and should be incorporated into the panel of mutations analyzed in high risk families of the appropriate ethnic background. Copyright 2001 S. Karger AG, Basel PMID- 11493754 TI - Community Genetics and Its Evaluation: A European Science Foundation Workshop. PMID- 11493755 TI - Analyses of matrix metalloproteinases and their inhibitors in cyst fluid of serous ovarian tumors. AB - OBJECTIVES: Expression of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) in serous tumors of the ovary was investigated to determine whether and how these proteolytic enzymes are associated with the progression of these tumors. METHODS: Cyst fluid of 24 serous ovarian tumors (8 adenocarcinomas, 2 borderline tumors and 14 adenomas) was analyzed using gelatin/casein zymography and enzyme-linked immunosorbent assay. RESULTS: Concentrations of MMP-9 and MMP-2 were statistically higher in serous adenocarcinomas than in serous adenomas (p < 0.01, p < 0.05, respectively), while the concentrations of TIMP-1 and TIMP-2 showed no significant difference between adenocarcinomas and adenomas. The molar ratio of TIMP-2/MMP-2 was lower in adenocarcinomas than in adenomas (p < 0.05). With gelatin zymography, the MMP-9 band was detected in all serous adenocarcinomas, but only in 8 of 14 serous adenomas (p = 0.05). Using casein zymography, MMP-7 was more frequently detected in serous adenocarcinomas (7/8) than in serous adenomas (4/14; p < 0.05). CONCLUSIONS: These observations indicate that matriolytic enzymes such as MMP-2, MMP-7 and MMP-9 are secreted into cyst fluid from serous adenocarcinoma tissues. In part, the aggressive invasion of serous carcinoma cells may be explained by the expression of matriolytic enzymes. PMID- 11493756 TI - CD50 and CD62L adhesion receptor expression on naive (CD45RA+) and memory (CD45RO+) T lymphocytes in the elderly. AB - A complex reshaping characterizes cellular immunity in the elderly. In particular, the hallmark of the "senescence" of the T cell compartment is a decrease in the proportion of CD45RA+ naive T lymphocytes concomitantly with an expansion of CD45RO+ memory T cells. However, in addition to age-dependent changes in their representation, phenotypical and functional anomalies also characterize naive and memory T cell populations in the elderly. Since cell adhesion molecules (CAMs) are multifunctional receptors which play important roles not only in cell-to-cell and cell-to-matrix interactions but also in signal transduction and cell activation, we analysed, by means of a three-colour flow cytometry method, the proportion, absolute number and density expression or mean fluorescence intensity (MFI) of CD50 (ICAM-3) and CD62L (L-selectin homing receptor) adhesion receptors on CD45RA+ and CD45RO+ peripheral blood CD3+ T cell subsets from 10 healthy elderly subjects and 10 young controls. Our aim was to investigate age-dependent changes in the expression pattern of these CAMs on naive and memory lymphocytes which might contribute to the remodelling of the immune system in the elderly. We considered the mean values +/- standard deviations of the percentage, absolute number and MFI of positive cells. The percentage of naive T cells expressing CD50 was not significantly modified in aged (94.8 +/- 5.0%) compared to young individuals (97.8 +/- 3.2%). On the contrary, the percentage of memory T cells exhibiting CD50 was lower in elderly than young donors (92.0 +/- 6.4 vs. 98.3 +/- 2.2%; p < 0.01). The percentage of naive T cells expressing CD62L was decreased in the elderly donors (53.3 +/- 18.8 vs. 80.8 +/- 11.0%; p < 0.001), whereas the proportion of CD62L+ memory T lymphocytes was substantially comparable between the two age groups (63.5 +/- 15.7 vs. 54.7 +/- 12.3%). The absolute number per mm(3) of CD50+ naive T cells from aged individuals was decreased (251.9 +/- 141.9 vs. 621.8 +/- 238.0/mm(3); p < 0.001), whereas memory peripheral blood T lymphocytes expressing CD50 were substantially unchanged (863.8 +/- 260.9 vs. 802.7 +/- 139.6/mm(3)). On the contrary, the absolute numbers per mm(3) of naive and memory peripheral blood T lymphocytes exhibiting CD62L were respectively decreased (190.8 +/- 133.4/mm(3)) and increased (515.1 +/- 146.8/mm(3)) in elderly donors compared to young controls (601.3 +/- 129.1 and 351.8 +/- 195.0/mm(3); p < 0.001 and p < 0.05, respectively). Finally, CD50 MFI values of naive as well as memory T cell subpopulations from aged subjects were increased compared to young donors (14.0 +/- 2.0 vs. 9.8 +/- 1.2 and 14.0 +/- 2.0 vs. 11.6 +/- 1.3; p < 0.001 and p < 0.01, respectively). CD62L was also overexpressed in both naive (8.4 +/- 1.6 vs. 6.7 +/- 1.4; p < 0.05) and memory (10.3 +/- 2.5 vs. 5.4 +/- 1.1; p < 0.001) T subsets in the elderly. CD50 and CD62L upregulation could be interpreted as a compensatory mechanism for a decreased responsiveness and a greater requirement for activation signals rather than an age-related anomaly. PMID- 11493757 TI - Primary thymic endocrine failure in HIV-1-infected children. AB - Thymulin, an essential hormone for the T lymphocyte differentiation process and function, was evaluated to asses thymic endocrine function in a cohort of 17 HIV 1-infected children aged between 2 months and 14 years, 18 seroreverted subjects and 47 normal controls. The rosette inhibition assay by Dardenne and Bach (1975) is the only method available to evaluate the biologically active form of this hormone (thymulin or Zn-facteur thymique serique, Zn-FTS), as immunoassays cannot discriminate between thymulin and the inactive form of the hormone not containing Zn (FTS). HIV-1 patients presented undetectable or significantly lowered plasma levels of thymulin. Plasma zinc levels were significantly reduced in patients although inactive, zinc-unbound thymulin molecules were not demonstrated. The investigation of inhibitory anti-thymulin molecules performed in all patients was negative. Thymulin titers did not correlate with CD4+ lymphocyte count at the different disease stages. This study suggests that a primary thymic endocrine deficiency is present in HIV children. The critical importance of these results in assessing disease progression and a potential therapeutic approach are discussed. PMID- 11493758 TI - Protective effects of vitamin E and selenium on the renal morphology in rats fed high-cholesterol diets. AB - The histopathological effects of cholesterol and the protective effects of vitamin E and selenium (Se) on renal histology were examined in Sprague-Dawley rats. Light-microscopic evaluation of the renal cortex revealed: glomerular fibrosis, cellular and mesangial proliferation, capillary obliteration and cholesterol crystals in the tubular lumina of the cholesterol-fed group. These results suggest that oxidated LDL (O-LDL) is a cytotoxic factor which stimulates mesangial cell and matrix proliferation. Ultrastructurally, small and large lipid vacuolization in intracapillary lumina, adhesion of epithelial foot processes, mesangial foam cells and polymorphonuclear leukocytes were seen in the cholesterol-fed group. In the groups fed cholesterol + vitamin E, cholesterol + Se and cholesterol + vitamin E + Se, morphological improvements were observed. It appeared that an excess in O-LDL, reactive oxygen species and growth factors might play an important role in the pathogenesis of glomerulosclerosis. In addition, it was concluded that antioxidant therapy may prevent LDL oxidation and generation of free radicals. PMID- 11493759 TI - Stimulation of glycosaminoglycan biosynthesis by umbilical cord serum of newborns delivered by mothers with EPH gestosis (preeclampsia). AB - OBJECTIVE: Edema, proteinuria, hypertension (EPH) gestosis, also known as preeclampsia, is the most common, pregnancy-associated pathological syndrome. It is accompanied by a significant increase in collagen content in the umbilical cord arteries and early replacement of hyaluronic acid by sulfated glycosaminoglycans (GAGs) both in these arteries and in Wharton's jelly. Such a remodelling of the umbilical cord tissues is accompanied by an increase in insulin-like growth factor-I (IGF-I) concentration in the umbilical cord serum. METHODS: Contact-inhibited human fibroblasts were incubated in Dulbecco culture medium containing control or EPH umbilical cord serum and supplemented with (14)C glucosamine or (35)S-sulfate. Radioactive GAGs were isolated, submitted to electrophoretic fractionation and quantified. RESULTS: The presence of umbilical cord serum in culture medium strongly stimulated the incorporation of (14)C glucosamine and (35)S-sulfate into GAGs synthesized by these cells. EPH serum was much more active in stimulation of sulfated GAGs biosynthesis than control serum, whereas the biosynthesis of hyaluronic acid was stimulated by both sera to a similar degree. CONCLUSION: Since IGF-I is known as a stimulator of collagen and sulfated GAG biosynthesis, the high concentration of this growth factor in the umbilical cord plasma may be responsible for preeclampsia-associated remodeling of the umbilical cord. PMID- 11493760 TI - The effect of renin-angiotensin axis inhibition on early atherogenesis in LDL receptor-deficient mice. AB - OBJECTIVE: The renin-angiotensin system may play a role in the development of atherosclerosis. Nevertheless, different results from studies attempting to attenuate the process by inhibiting the converting enzyme were equivocal, and in those who succeeded, blood pressure was lowered and/or the lipid profile was improved in addition to the inhibition of the renin-angiotensin axis. The aim of this study is to investigate the effect of low doses of fosinopril, a converting enzyme inhibitor, on the development of atherosclerosis in LDL-receptor-deficient mice. METHODS: Three groups of 15 mice were fed a high-fat, high-cholesterol western diet. The three study groups received either distilled water (control group), or water supplemented with fosinopril 0.01 mg/kg/day (low-dose group) or with 0.1 mg/kg/day (high-dose group). Plasma aldosterone levels and lipid profiles were measured at the beginning and at the end of the study. After 10 weeks, the mice were sacrificed and the extent of atherosclerosis was assessed at the aortic sinus. RESULTS: Plasma aldosterone levels did not change in the control group, but decreased significantly in both treated groups from 74.7 to 39.3 ng/ml in the low-dose group (p < 0.003) and from 70.7 to 33.6 ng/ml in the high-dose group (p < 0.001). The lipid profile at the end of the study showed significantly lower levels of cholesterol and triglycerides in the high-dose group as compared to the low-dose group (p < 0.05). There was no difference between the three groups regarding the area of atherosclerosis at the aortic sinus: 157,000 +/- 34,000, 130,000 +/- 58,000 and 145,000 +/- 26,000 microm(2) in the control, low-dose and high-dose groups, respectively. CONCLUSION: Inhibition of the renin-angiotensin-aldosterone axis by itself does not prevent the development of early atherosclerosis in LDL-receptor-deficient mice. PMID- 11493761 TI - Ontogeny of pig discrete Peyer's patches: distribution and morphometric analysis. AB - We investigated the development of lymphoid and non-lymphoid cells in discrete Peyer's patches (PP) of the pig using immuno-histology and image analysis. In newborn piglets discrete PP were mainly populated by CD2+, CD3+ T cells, and major histocompatibility complex class II+ cells, many of which were of macrophage and dendritic cell lineage. Four days after birth, cells were localized in defined regions: the follicle; the inter-follicular area and the dome region. Compartmentalization within the follicle started about 6 days after birth. The first signs of secondary follicles were seen from about 14 days. The pig discrete PP attained their mature structure at about 3 weeks after birth. Here we show that despite the demonstration at birth of the cell types that support antigen processing and presentation, PP did not fully differentiate morphologically until at least this time when antigen can be handled in an efficient manner. PMID- 11493762 TI - Investigations into the molecular effects of single nucleotide polymorphism. AB - OBJECTIVES: DNA sequences are very rich in short repeats and their pattern can be altered by point mutations. We wanted to investigate the effect of single nucleotide polymorphism (SNP) on the pattern of short DNA repeats and its biological consequences. METHODS: Analysis of the pattern of short DNA repeats of the Thy-1 sequence with and without SNP. Searching for DNA-binding factors in any region of significance. RESULTS: Comparing the pattern of short repeats in the Thy-1 gene sequences of Turkish patients with ataxia telangiectasia (AT) with the "wild type" sequence from the DNA database, we identified a missing 8-bp repeat element due to an SNP in position 1271 (intron II) in AT-DNA sequences. Only the mutated sequence had the potential for the formation of a stem loop in DNA or pre mRNA. In super-shift experiments we found that DNA oligomers covering the area of this SNP formed a complex with proteins amongst which we identified the proliferating cell nuclear antigen (PCNA) protein. CONCLUSION: SNPs have the potential to alter DNA or pre-mRNA conformation. Although no SNP-depeding formation of the DNA-protein complex was evident, future investigations could reveal differential molecular mechanisms of cellular regulation. PMID- 11493763 TI - Expression of lymphotoxin-alpha by keratinocytes: a further mediator for the lichenoid reaction. AB - OBJECTIVE: Lichen planus (LP) represents a disease in which autoimmune mechanisms mediated by Th1 T cells are involved. Lymphotoxin-alpha (LT-alpha) represents a Th1 cytokine with proinflammatory activities in LP, as has recently been demonstrated for interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). METHODS: Expression of LT-alpha mRNA was investigated by RT-PCR and nonradioactive in situ hybridization. Double staining methods were applied to characterize the phenotype of cells expressing LT-alpha. Cell stimulation experiments were performed on the transformed squamous cell line HaCaT. RESULTS: In contrast to normal skin, LT-alpha-specific RT-PCR products were found in all cases of LP. Cells in the inflammatory infiltrate expressing LT-alpha were identified as mainly T cells and mast cells, as shown by in situ hybridization. Furthermore, predominant LT-alpha mRNA expression could be observed in lesional keratinocytes adjacent to the band-like inflammatory infiltrate. In cell stimulation experiments, it could be shown that IFN-gamma induces LT-alpha and TNF-alpha mRNA in the human squamous cell line HaCaT, concomitant with upregulation of MHC class II and intercellular adhesion molecule-1, which could also be observed on lesional keratinocytes in LP. CONCLUSIONS: In LP, LT-alpha mRNA is predominantly expressed by lesional keratinocytes and to a lesser extent by inflammatory cells. Induction of LT-alpha in keratinocytes is closely related to the expression of TNF-alpha and MHC class II. The loci of TNF-alpha and LT alpha map to MHC class III on chromosome 6, which is closely linked to the MHC class II gene locus. Our results suggest that stimulation of keratinocytes with IFN-gamma results in the upregulation of proinflammatory cytokines such as LT alpha and TNF-alpha as well as MHC class II, which map to the same gene region of immunoregulatory genes on chromosome 6 and may be involved in the induction and maintenance of the disease. PMID- 11493765 TI - Fat reduction to prevent prostate cancer: waiting for more evidence? AB - Complementary medicine has become an important area of interest for patients and researchers around the world. The utilization of some of these therapies by many individuals makes it imperative to understand whether they have any role in treatment of cancer or other diseases. Some of such therapies may play a role in the prevention of prostate cancer. Clinical trials are addressing this issue, and whether these products could also improve prognosis of prostate cancer. That dietary fat reduction may help to prevent prostate cancer is supported by numerous case-control studies reported over the past several decades, but recent prospective studies suggest that the impact of fat reduction in this regard may not be great. Clinicians should be careful not to suggest such a benefit until more research provides a better picture of the situation. Breast cancer is probably the best example of why more research into dietary changes for reduction in cancer risk is needed. Once believed to play a significant role, recent prospective studies suggest that dietary fat reduction may have little effect on breast cancer prevention, although some interesting insights have been gained with regard to the method of meat preparation and the influence of genetics. Fat reduction, together with soy products or other plant estrogen foods, may have a symbiotic relationship. Numerous healthy lifestyle changes incorporated and practiced at one time (healthy diet, attainment and maintenance of normal weight, soy consumption, among others) may hold some promise in the area of cancer prevention. In the meantime any healthy lifestyle or dietary change should be encouraged, because it may reduce the risk of cardiovascular disease, which is still the number one cause of mortality. It is also an important cause of morbidity and mortality in cancer patients. Regardless, complementary medicine should probably be discussed with any patient who is initiating or undergoing conventional treatment, because of the cardiovascular benefits and overall potential impact on all-cause mortality. Whether such therapies impact on prostate carcinoma remains to be determined through additional prospective investigations. PMID- 11493766 TI - The genetic basis of renal epithelial tumors: advances in research and its impact on prognosis and therapy. AB - The genetics of renal cell carcinoma continues to elucidate the pathways of kidney tumorigenesis. The relationship between the VHL gene and clear cell carcinoma, MET and papillary carcinoma, and the families of genes that they regulate, continues to be unraveled. New hereditary kidney cancer syndromes, like familial oncocytoma and the Birt-Hogg-Dube syndrome, have been identified and the search for the genes that cause them is under way. Researching the genetics of these disorders is essential for an understanding of sporadic kidney cancer genetics. This chapter will review the current knowledge of the hereditary kidney cancer syndromes, the genes that cause them, new advances in genetic research and techniques, and how this information impacts upon diagnostic, prognostic, and therapeutic methods of the future. PMID- 11493767 TI - Ultrasound for prostate imaging and biopsy. AB - Transrectal ultrasound guided systemic sextant needle biopsy of the prostate has been the procedure of choice for the diagnosis of prostate cancer. Several shortcomings of this procedure have been recognized and there is concern that it may represent an inadequate sampling of the prostate. Refinements include modifications of biopsy location and an increase in the number of cores obtained. Enhanced ultrasound techniques may improve the accuracy of prostate biopsy. In addition, research continues to develop prognostic factors derived from the core biopsy that may enhance the prediction of tumor biology. This paper provides a basic review of transrectal ultrasound diagnosis of prostate cancer with emphasis on advances in this area. PMID- 11493768 TI - Laparoscopic surgery in urological pelvic cancer. AB - Laparoscopy has become more widely used in urological practice, both for upper and lower urinary tracts. Improvements in instrumentation, and in surgeons' skills and experience have broadened the applications of laparoscopy. In particular, indications for laparoscopy in urological pelvic oncology have emerged as possible but still theoretical alternatives to conventional surgery. The development of the laparoscopic approach is still under close evaluation. Beyond these 'conventional' forms of laparoscopy, potential technical applications of computer assistance provide a glimpse of what could become the surgery of tomorrow. PMID- 11493769 TI - Mechanisms governing bone metastasis in prostate cancer. AB - Why does prostate cancer metastasize to bone? Why is there increased turnover of the bone matrix in the presence of prostate cancer? A recent autopsy study supports a traditional hypothesis that gross, anatomic patterns of blood flow influence the distribution of metastatic deposits. On the other hand, recent developments in animal models of prostate cancer bone metastasis have renewed interest in the traditional 'seed and soil' hypothesis: several studies point to specific biological interactions between prostate cancer cells and the bone microenvironment that can explain the tendency of prostate cancer cells to colonize bone and grow into clinically relevant metastatic deposits. Some studies implicate mechanisms including chemoattraction and enhanced adherence to bone endothelium. Additional data suggest that prostate cancer cells are 'osteomimetic', that is, they take on the properties and behaviors of osteoblasts or osteoclasts upon arrival in bone. These activities lead to enhanced turnover of the bone matrix and may explain the propensity of prostate cancer to grow in bone. Finally, a series of studies have implicated other molecular markers as distinguishing characteristics of bone-metastatic prostate cancer tissue. PMID- 11493770 TI - The prospect of gene therapy for prostate cancer: update on theory and status. AB - Molecularly based novel therapeutic agents are needed to address the problem of locally recurrent, or metastatic, advanced hormone-refractory prostate cancer. Recent basic science advances in mechanisms of gene expression, vector delivery, and targeting have rendered clinically relevant gene therapy to the prostatic fossa and distant sites feasible in the near future. Current research and clinical investigative efforts involving methods for more effective vector delivery and targeting, with enhanced gene expression to selected (specific) sites, are reviewed. These areas of research involve tissue-specific promoters, transgene exploration, vector design and delivery, and selective vector targeting. The 'vectorology' involved mainly addresses selective tissue homing with ligands, mechanisms of innate immune system evasion for durable transgene expression, and the possibility of repeat administration. PMID- 11493771 TI - Allogeneic stem cell transplantation for renal cell carcinoma. AB - Although the prognosis for patients with metastatic kidney cancer remains poor, a number of promising immunotherapeutic approaches for the treatment of metastatic disease have been developed over the past decade. The response of some patients to cytokines such as interleukin-2 and interferon-alpha, and more recently, vaccination with dendritic cell/tumor fusions has laid the ground work for ongoing immune-based investigational approaches. Allogeneic stem cell transplantation is a potent form of immunotherapy capable of delivering potentially curative immune-mediated anti-tumor effects against a number of different hematological malignancies. Knowledge of renal cell carcinoma's unusual susceptibility to immune attack has led to the hypothesis that tumor rejection, mediated through immunocompetent donor T-cells, might be generated against this solid tumor following the transplantation of an allogeneic immune system. Although clinical trials are early and ongoing, the recent observation of metastatic disease regression following non-myeloablative stem cell transplantation has identified renal cell carcinoma as being susceptible to a graft-versus-tumor effect. Disease responses following such therapy have ranged from partial to complete and have been observed even in patients who have failed conventional cytokine based strategies. This article reviews the design, methodology and early clinical results of studies investigating the use of allogeneic stem cell transplantation in metastatic renal cell carcinoma. PMID- 11493772 TI - Can biological markers replace cystoscopy? An update. AB - Cystoscopy is currently considered the gold standard for the detection of bladder tumors. The role of urine cytology in the initial detection and follow-up of patients is under discussion. New elaborative and rapid assays are available that may circumvent the low sensitivity and poor reproducibility of urine cytology. The methods that have been tested extensively are the nuclear matrix protein (NMP22) assay, the BTA stat assay, and the BTA TRAK enzyme-linked immunosorbent assay. Both outperform cytology in the detection of low-grade lesions. The specificity of both assays, however, lags behind that of cytology. The data from retrospective analyses are insufficient to justify clinical integration, and the need to replace cystoscopy with these novel assays remains to be proven. PMID- 11493773 TI - Recent clinical trials in superficial bladder cancer. AB - The present review addresses literature regarding the management of superficial bladder cancer published since March 2000. There is no definitive winner among urinary markers of bladder cancer, because they lack specificity or are insufficiently tested. Pathologists continue in their efforts to improve prediction of evolution of superficial bladder cancer to recurrent or infiltrative disease. A few studies have confirmed the value of previously described prognostic factors for recurrence and progression, and have added some refinements. Transurethral resection is not as complete as was believed. Fluorescence detection of flat bladder carcinoma has been demonstrated to improve diagnosis and treatment. The necessity to perform a repeat transurethral resection in high-grade superficial bladder cancer became evident. Identification of the working mechanisms of bacille Calmette-Guerin on superficial bladder cancer remains an important objective, and may help to improve treatment schedules and avoid the morbidity associated with bacille Calmette-Guerin administration. Patients who are at high risk may benefit from long-term maintenance bacille Calmette-Guerin therapy. Valrubicin and keyhole limpet haemocyanin appear to be promising agents in the treatment of superficial bladder cancer. PMID- 11493774 TI - New chemotherapy combinations for advanced bladder cancer. AB - Transitional cell carcinoma of the urothelium is considered a chemosensitive malignancy. Until recently, the methotrexate, vinblastine, doxorubicin and cisplatin combination has been considered the standard for treating this disease. The development of new chemotherapeutic agents such as gemcitabine and the taxanes has opened up promising new perspectives in the treatment of this disease. However, the preliminary phase II data must be confirmed in adequately conducted phase III trials. PMID- 11493775 TI - Second-line treatment of advanced transitional cell carcinoma of the urothelial tract. AB - Cisplatin-based combination chemotherapy such as methotrexate, vinblastine, adriamycin and cisplatin produces durable improvements in survival in only a minority of patients. Therefore, other therapeutic options and strategies are clearly needed. Strategies include increasing the dose of chemotherapy, modifying the sequencing of chemotherapy, and new therapeutic agents. This paper reviews recent work on high-dose chemotherapy, currently available chemotherapeutic agents and combinations, with an emphasis on gemcitabine and the taxanes. New strategies such as monoclonal antibody therapy and molecular targeted small molecule therapy are becoming a reality in the treatment of many diseases. The rationale for using epidermal growth factor receptor targeted therapies is also reviewed. PMID- 11493776 TI - Prognostic factors in clinical stage 1 non-seminomatous testicular tumours. AB - For patients with a clinical stage 1 non-seminomatous germ cell tumour of the testis cure rates should be close to 100%, whether surveillance, primary surgery, primary chemotherapy or a combination is chosen. The identification of patients with microscopic metastases is difficult. Even with the best predictive factors currently available (vascular invasion and percentage embryonal cell carcinoma in the primary tumour), the identification of micro-metastases is no better than the flip of a coin. Several additional prognostic factors have been studied, but none is yet applicable in daily practice. PMID- 11493777 TI - Leadership, professional heroism, and the Eastern Association for the Surgery of Trauma: presidential speech at the 14th scientific assembly. PMID- 11493778 TI - ARDS after pulmonary contusion: accurate measurement of contusion volume identifies high-risk patients. AB - BACKGROUND: The pathophysiology of pulmonary contusion (PC) is poorly understood, and only minimal advances have been made in management of this entity over the past 20 years. Improvement in understanding of PC has been hindered by the fact that there has been no accurate way to quantitate the amount of pulmonary injury. With this project, we examine a method of accurately measuring degree of PC by quantifying contusion volume relative to pulmonary function and outcome. METHODS: Patients with PC from isolated chest trauma who had admission chest computed tomographic scan were identified from the registry of a Level I trauma center over a 1.5-year period. Subsequently, prospective data on all patients admitted to the intensive care unit with PC during a 5-month period were collected and added to the retrospective database. Using computer-generated three-dimensional reconstruction from admission chest computed tomographic scan, contusion volume was measured and expressed as a percentage of total lung volume. Admission pulmonary function variables (Pao2/FiO2, static compliance), injury descriptors (chest Abbreviated Injury Score, Injury Severity Score, injury distribution), and indicators of degree of shock (admission systolic blood pressure, admission base deficit) were documented. Outcomes included maximum positive end-expiratory pressure, ventilator days, pneumonia, and acute respiratory distress syndrome (ARDS). RESULTS: Forty-nine patients with PC (35 bilateral) were identified. The average severity of contusion was 18% (range, 5-55%). Patients were classified using contusion volume as severe PC (> or =20%, n = 17) and moderate PC (< 20%, n = 32). Injury Severity Score was similar in the severe and moderate groups (23.3 vs. 26.5, p = 0.33), as were admission Glasgow Coma Scale score (12 vs. 13, p = 0.30), admission blood pressure (131 vs. 129 mm Hg, p = 0.90), and admission Pao2/Fio2 (197 vs. 255, p = 0.14). However, there was a much higher rate of ARDS in the severe group as compared with the moderate group (82% vs. 22%, p < 0.001). There was a trend toward higher pneumonia rate in the severe group, with 50% of patients in the severe group developing pneumonia as compared with 28% in the moderate group (p = 0.20). CONCLUSION: Extent of contusion volumes measured using three-dimensional reconstruction allows identification of patients at high risk of pulmonary dysfunction as characterized by development of ARDS. This method of measurement may provide a useful tool for the further study of PC as well as for the identification of patients at high risk of complications at whom future advances in therapy may be directed. PMID- 11493779 TI - Pediatric minor head trauma: indications for computed tomographic scanning revisited. AB - BACKGROUND: Although the use of computed tomographic (CT) scanning in severe head trauma is an accepted practice, the indications for its use in minor injury remain ill defined and subjective. We sought to define the incidence and identify risk factors for intracranial injury (ICI) after minor head trauma in children who did not have suspicious neurologic symptoms in the field or on presentation. METHODS: From January 1, 1992, until April 1, 2000, 569 blunt trauma patients (age < 16 years) with a Glasgow Coma Scale score of 14 or 15 triaged by American College of Surgeons Pediatric Mechanism Criteria at a Level I trauma center received head CT scan. Loss of consciousness (LOC) status was known for 429. This subgroup was retrospectively reviewed for mechanism, age, Injury Severity Score, LOC status, GCS score, associated injuries, and CT scan findings (normal, fracture only, or intracranial injury). Relative risk values for intracranial injury were generated and statistical significance was assessed. RESULTS: Fourteen percent (62 of 429) of study patients (GCS score of 14 and 15) had ICI. Sixteen percent of patients (35 of 215) with GCS score of 15 and (-)LOC (negative for LOC) had intracranial injury manifesting as subdural hematoma, epidural hematoma, subarachnoid hemorrhage, or brain contusion. Three required surgery for intracranial mass lesions. One patient deteriorated and required intubation and intensive care unit management. Neither (+)LOC (positive for LOC) nor GCS score of 14 increased the likelihood of intracranial injury over those patients without loss of consciousness or with GCS score of 15. Distant injury was also not an independent predictor of ICI for those with GCS scores of 14 or 15, as 84% of the ICI group had head injury only. Skull fracture was a risk factor for ICI but had poor negative predictive value, as 45% of patients with ICI did not have fractures. Similarly, minor craniofacial soft tissue trauma was a significant risk factor (relative risk, 11) that had marginal negative predictive value (0.95), as 14% (9 of 62) of ICI patients did not have superficial craniofacial injury. CONCLUSION: A normal neurologic exam and maintenance of consciousness does not preclude significant rates of intracranial injury in pediatric trauma patients. Contrary to convention, neither LOC nor mild altered mentation is a sensitive indicator with which to select patients for CT scanning. Skull fractures and superficial craniofacial injury are similarly unreliable. Identification of these patients is important for the occasional case requiring intervention and for the tracking of complications. A liberal policy of CT scanning is warranted for pediatric patients with a high-risk mechanism of injury despite maintenance of normal neurologic status in the field and at hospital screening. PMID- 11493780 TI - Macrophage cyclooxygenase expression, immunosuppression, and cardiopulmonary dysfunction after blunt chest trauma. AB - BACKGROUND: Two series of experiments were performed in swine who received severe blunt chest trauma. The goals were to determine the time course of constitutive and inducible cyclooxygenase (COX) isozyme expression in pulmonary macrophages (Mphis), and to determine whether COX expression and cardiopulmonary dysfunction were altered when neutrophils (PMNs) were pharmacologically depleted with cyclophosphamide (CYC). METHODS: In series 1 (n = 17), anesthetized, mechanically ventilated swine were subjected to right chest trauma via captive bolt gun, hemorrhage, and a 60-minute shock period. In series 2 (n = 41), CYC (50 mg/kg intravenously) was administered 4 days before trauma, and the shock period was shortened to 30 minutes. In both series, hemodynamic support and supplemental oxygen were provided for an additional 60 to 90 minutes after shock. Mphis were isolated from serial bilateral bronchoalveolar lavages (BALs) and COX protein expression was measured with Western blots. RESULTS: In series 1, death occurred in 11 of 17. In survivors, Mphi COX-1 peaked at > 100 times baseline in both right BAL and left BAL by 60 minutes (before resuscitation). Changes in Mphi COX 2 were minimal. In series 2, before trauma, CYC (n = 16) reduced circulating and BAL PMNs by > 90% relative to control (n = 25, both p < 0.05) with no complicating side effects. After trauma, death occurred in 11 of 25 controls versus 9 of 16 with CYC. In survivors, PaO2/FIO2 was < 250 and PaCO2 was 25% higher on constant minute ventilation, indicating mismatched ventilation/perfusion; both changes were reduced with CYC (p < 0.05). In controls, bilateral histologic damage included edema, alveolar hemorrhage, and interstitial infiltrates. These changes were reduced by one third with CYC (p = 0.08). Trauma-induced changes in BAL protein, BAL elastase, or Mphi COX expression were not lessened by CYC. CONCLUSION: After unilateral chest trauma, Mphi COX-1, not COX-2, is induced bilaterally and before fluid resuscitation; CYC prevented PMN infiltration and attenuated structural and functional changes after resuscitation, which suggests that PMNs have a role in the pathogenic mechanism of secondary lung injury; Mphi COX expression and other injury markers were not altered by CYC; and since Mphis continued to express proinflammatory COX protein even after pretreatment with a powerful nonspecific immunosuppressant, and since there is residual alveolar capillary damage even in the absence of PMNs, it is logical to conclude that no single cell type or mediator is a practical therapeutic target and that novel resuscitation strategies must address multiple elements in the inflammatory cascade. PMID- 11493781 TI - Intraperitoneal blood exacerbates the remote inflammatory response to murine peritonitis. AB - BACKGROUND: This study investigated the effects of intra-abdominal blood on the systemic response to peritonitis using a murine model of hemorrhage, peritonitis, and multiple organ dysfunction syndrome. METHODS: The model used male ICR mice subjected to hemorrhage and intraperitoneal zymosan. Half of the mice received intraperitoneal blood. Outcome measures included lung myeloperoxidase, lung edema, lung injury score, and plasma and lung tissue chemokine production. RESULTS: Peritoneal blood (in association with peritoneal inflammation) increased lung neutrophil sequestration (myeloperoxidase) (2.56 +/- 1.42 vs. 1.45 +/- 0.49 U/left lung, p = 0.04) and lung weight (0.11 +/- 0.04 vs. 0.07 +/- 0.02 g/left lung, p = 0.02), and was associated with significantly higher chemokine levels in plasma (KC and MCP-1) and lung tissue (KC, MIP-2, and MCP-1). Both plasma and lung tissue neutrophil chemoattractants KC and MIP-2 were significantly linearly correlated with myeloperoxidase (p < 0.009), and lung tissue KC (a neutrophil chemokine) and MCP-1 and MIP-1alpha (mononuclear cell chemokines) correlated with lung injury score (p < 0.003). CONCLUSION: Although blood alone in the peritoneal cavity was well tolerated, in conjunction with inflammation, it was synergistic in amplifying the systemic inflammatory response. The amplified lung injury in this model was associated with significant increases in circulating and lung tissue chemokine concentrations. PMID- 11493783 TI - Evolution in damage control for exsanguinating penetrating abdominal injury. AB - OBJECTIVE: Damage control (DC) has proven valuable in exsanguinated patients. The purpose of this study was to quantify and qualify the impact of current damage control principles applied in a penetrating abdominal injury (PAI) population. METHODS: Over a 3-year period (June 1997-May 2000), of 271 laparotomies for PAI, 24 patients underwent DC (8.9%). Demographics, injury grade, resuscitative and operative parameters, acid-base status, coagulation profiles, fluid/transfusion requirements, definitive repairs, abdominal closure, complications, and outcomes were reviewed. Data were compared with our DC experience a decade earlier. Fisher's exact test was used for comparisons. RESULTS: Overall survival improved for equivalent Injury Severity Score, Revised Trauma Score, TRISS, admission systolic blood pressure, operating room systolic blood pressure, and Penetrating Abdominal Trauma Index score. Solids (1.2 vs. 1.3), hollow organ (1.5 vs. 1.7), and major vascular injuries (0.5 vs. 0.8) per patient remain unchanged. Currently, there was less hypothermia with equivalent operating room times. In intensive care unit survivors, acid-base status was similar but coagulopathy and hypothermia were less severe. Definitive colon management has shifted from ostomies to anastomoses. Eventual fascial closure occurred in 14 of 19 (74%) compared with 12 of 14 (86%) in the historical group. There were three gastrointestinal fistulae (one pancreatic), one anastomotic leak, and three intra abdominal abscesses. CONCLUSION: Continued application of DC principles has led to improved survival with PAI. Better control of temperature, experience with the open abdomen, and intensive care unit care may be causative. PMID- 11493784 TI - Implications of the "contrast blush" finding on computed tomographic scan of the spleen in trauma. AB - BACKGROUND: The "contrast blush" (CB) computed tomographic (CT) scan finding has often been used clinically as an indicator for therapeutic splenic intervention (SI) (splenectomy, splenorrhaphy, or angiographic embolization). We sought to examine the prognostic significance of this finding. METHODS: The records and CT scans of 324 trauma patients from two Level I trauma centers who had blunt splenic injury and a CT scan of the abdomen within 24 hours of admission were reviewed and screened for CB. RESULTS: CB was identified in 11% of patients, and its incidence was significantly related to the grade of injury: grade I/II, 3.2%; grade III, 11.8%; and grade IV/V, 26.3% (p < 0.001). SI was also related to the grade: grade I/II, 7.7%; grade III, 37.6%; and grade IV/V, 69.7% (p < 0.001). The chance of having SI was greater in those with CB (75.0%) when compared with those without CB (25.0%) (p < 0.001; odds ratio, 9.2). A multivariate logistic regression analysis revealed that SI correlated independently with splenic grade, emergency department hypotension, and age, but did not demonstrate a correlation with CB. CONCLUSION: CB is not an absolute indication for an operative or angiographic intervention. Factors such as patient age, grade of injury, and presence of hypotension need to be considered in the clinical management of these patients. PMID- 11493785 TI - Blunt cerebrovascular injuries: diagnosis and treatment. AB - BACKGROUND: Blunt cerebrovascular injuries (BCVIs), once thought to be rare, have been recognized with increasing frequency in recent years. An incidence of 0.33% for carotid artery injury (CAI) was noted from our institution, with 24% stroke related mortality. Vertebral artery injury (VAI) has been thought both rare and of questionable significance. Incidence, associated injury patterns, and outcomes were examined during a period of aggressive screening (four-vessel angiography). METHODS: Patients with BCVI were identified from the registry of a Level I trauma center over a 5-year period (1995-1999). RESULTS: One hundred thirty-nine BCVIs were identified in 96 patients: 75 were CAIs (14 bilateral), 64 were VAIs (14 bilateral), and 15 patients had both CAI and VAI. The incidence of CAI was 0.5% of all blunt trauma admissions, significantly higher than our earlier experience (p < 0.0002), whereas that for VAI was 0.4%. Thirty-four percent of CAIs were diagnosed because of ischemic changes and 38% because of injury pattern (neck, Horner syndrome, basilar skull fracture); only 12% of VAIs were diagnosed because of posterior circulation ischemia, with 64% because of injury pattern (cervical spine, soft tissue, facial fracture). Stroke-related mortality for CAI was 13%, and that for VAI was 4%. Forty-three of the 75 CAIs were treated (anticoagulation/antiplatelet) before development of ischemia. Thirty-nine of the 50 VAI patients were treated before development of ischemia. Stroke rate for CAI was 31% (hemiplegia/hemiparesis) and for VAI was 14% (brain stem dysfunction). Stroke rate for treated vessels (heparin, antiplatelet therapy) with CAI was 6.8% compared with 64% in untreated vessels (p < 0.001). Treated patients with VAI had a stroke rate of 2.6%, whereas untreated patients developed stroke 54% of the time. CONCLUSION: Increased awareness and aggressive screening have resulted in significantly increased incidence of diagnosis of CAI, with associated lower stroke-related mortality. VAIs have been noted with similar incidence, and though the stroke rate is lower with VAI, stroke outcomes are generally catastrophic. Anticoagulation therapy is effective for both varieties of BCVI. PMID- 11493786 TI - Perceived threat to life predicts posttraumatic stress disorder after major trauma: risk factors and functional outcome. AB - BACKGROUND: The importance of psychological morbidity after major trauma, such as posttraumatic stress disorder (PTSD), is continuing to gain attention in trauma outcomes research. The Trauma Recovery Project is a large prospective epidemiologic study designed to examine multiple outcomes after major trauma, including quality of life (QoL) and PTSD. The specific objectives of the present report are to examine risk factors for PTSD and to assess the impact on QoL at the 6-, 12-, and 18-month follow-up time points in the Trauma Recovery Project population. METHODS: Between December 1, 1993, and September 1, 1996, 1,048 eligible trauma patients triaged to four participating trauma center hospitals in the San Diego Regionalized Trauma System were enrolled in the study. The enrollment criteria for the study included age 18 years and older, admission Glasgow Coma Scale score of 12 or greater, and length of stay > 24 hours. QoL was measured after injury using the Quality of Well-being scale, a sensitive index to the well end of the functioning continuum (range, 0 = death to 1.000 = optimum functioning). Early symptoms of acute stress reaction (SASR) at discharge were assessed using the Impact of Events Scale (score > 30 = SASR). PTSD at 6-month follow-up was diagnosed using standardized Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, criteria. Patient outcomes were assessed at discharge and at 6, 12, and 18 months after discharge. RESULTS: PTSD was diagnosed in 32% (261 of 824) patients at 6-month follow-up. Perceived threat to life predicted PTSD onset (odds ratio [OR], 1.6; p < 0.01) and early SASR (OR, 2.2; p < 0.001). PTSD was more frequent in women (39%) than in men (29%) and in younger low-income patients. Other major risk factors were penetrating trauma (OR, 2.3; p < 0.001) and assaults (OR, 1.5; p < 0.05). PTSD had a major impact on QoL at 6-, 12-, and 18-month follow-up (Quality of Well-being scale score: 6 months, 0.576 vs. 0.658; 12 months, 0.620 vs. 0.691; 18 months, 0.620 vs. 0.700; p < 0.0001). CONCLUSION: These results provide new and provocative evidence that perceived threat to life and mechanism predict PTSD after major traumatic injury. PTSD had a prolonged and profound impact on short- and long-term outcome and QoL. PMID- 11493787 TI - Preparation and achievement of American College of Surgeons level I trauma verification raises hospital performance and improves patient outcome. AB - OBJECTIVE: The purpose of this study was to assess the impact on patient outcome and hospital performance of preparing for and achieving American College of Surgeons (ACS) Level I trauma verification. METHODS: The center was a previously designated state regional trauma center located adjacent to a major metropolitan area. Preparation for ACS verification began in early 1996 and was completed in early 1998. Final verification took place in April 1999. Data were analyzed before (1994) and after (1998) the process. There was a marked increase in administrative support with trauma named one of the hospital's six centers of excellence. Two full-time board-certified trauma/critical care surgeons were added to the current six trauma surgeons. Their major focus was trauma care. Trauma support staff was also increased with case managers, a trauma nurse practitioner, additional trauma registrars, and administrative support staff. Education and continuous quality improvement were markedly expanded starting in 1996. RESULTS: There were 1,098 trauma patients admitted in 1994, and 1,658 in 1998. Overall mortality decreased (1994, 7.38%; 1998, 5.37%; p < 0.05). There was a marked decrease in mortality for severely injured (Injury Severity Score > 30) patients (1994, 44% mortality [38 of 86]; 1998, 27% [22 of 80]; p < 0.04). Average length of stay also decreased (1994, 12.22 days; 1998, 9.87 days; p < 0.02). This yielded an estimated cost savings for 1998 of greater than $4,000 per patient (total saving estimate of $7.4 million). CONCLUSION: Trauma system improvement as related to achieving ACS Level I verification appeared to have a positive impact on survival and patient care. There were cost savings realized that helped alleviate the added expense of this system improvement. The process of achieving ACS Level I verification is worthwhile and can be cost effective. PMID- 11493788 TI - Public hospital-based level I trauma centers: financial survival in the new millennium. AB - BACKGROUND: The medical benefits of trauma centers have been well documented; studies have reported substantial financial losses attributed to trauma care. This study demonstrates the dependence of Level I trauma centers on Disproportionate Share Hospital (DSH) governmental funds and tax dollars. Furthermore, specific injury groups have greater dependence on these funds. METHODS: Records of 553 trauma patients admitted to a public urban Level I trauma center during a 6-month period were reviewed. Patients were grouped according to blunt, penetrating, and thermal injuries. Data for each group included charges, costs, payments, and the source of reimbursement. Profit and loss margins were compared with and without government funds. RESULTS: With diminished DSH funds and tax dollars, a net loss over $2.1 million was incurred. The greatest disparity originates from Medicaid, self-pay, and prisoner patient groups. Inclusion of government funds provided a positive return of over $600,000. CONCLUSION: The financial stability of urban public Level I trauma centers without additional funding is tenuous because of a high proportion of uninsured and underinsured patients. Government tax dollars and DSH funds are required for their continued solvency. PMID- 11493789 TI - Liberalized screening for blunt carotid and vertebral artery injuries is justified. AB - BACKGROUND: Current literature suggests that blunt carotid injuries (BCIs) and vertebral artery injuries (BVIs) are more common than once appreciated. Screening criteria have been suggested, but only one previous study has attempted to identify factors that predict the presence of BCI/BVI. This current study was conducted for two reasons. First, we wanted to determine the incidence of BCI/BVI in our institution. Second, we wanted to determine the incidence of abnormal four vessel cerebral angiograms ordered for injuries and signs believed to be associated with BCI/BVI and thus to determine whether the screening protocol developed was appropriate. METHODS: From August 1998, we used liberalized screening criteria for patients who were prospectively identified and suspected to be at high risk for BCI/BVI if any of the following were present: anisocoria, unexplained mono-/hemiparesis, unexplained neurologic exam, basilar skull fracture through or near the carotid canal, fracture through the foramen transversarium, cerebrovascular accident or transient ischemic attack, massive epistaxis, severe flexion or extension cervical spine fracture, massive facial fractures, or neck hematoma. Four-vessel cerebral angiograms were used for screening for BCI/BVI. RESULTS: Over the 18-month study period, 48 patients were angiographically screened, with 21 patients (44%) being identified as having a total of 19 BCIs and 10 BVIs. Nine patients had unilateral carotid artery injuries and three patients had bilateral carotid artery injuries. Vertebral artery injuries were unilateral in six patients. One patient had bilateral carotid artery injuries and a unilateral vertebral artery injury. One patient had a unilateral carotid artery injury and a unilateral vertebral artery injury, and one patient had a unilateral carotid artery injury and bilateral vertebral artery injuries. During the same study period, 2,331 trauma patients were admitted, with 1,941 (83%) secondary to blunt trauma. The overall incidence of BCI/BVI was 1.1%. The frequency of abnormal angiograms ordered for cerebrovascular accident or transient ischemic attack, massive epistaxis, or severe cervical spine fractures was 100%. The frequency of abnormal angiograms ordered for the other indications was as follows: fracture through foramen transversarium, 60%; unexplained mono- or hemiparesis, 44%; basilar skull fracture, 42%; unexplained neurologic examination, 38%; anisocoria, 33%; and severe facial fractures, 0%. CONCLUSION: The liberalized screening criteria used in this study were appropriate to identify patients with BCI/BVI. This study suggests BCI/BVI to be more common than previously believed and justifies that screening should be liberalized. PMID- 11493791 TI - Computed tomographic scan can be used for surgical decision making in zone II penetrating neck injuries. AB - BACKGROUND: Controversy surrounds the evaluation of zone II penetrating neck injuries. Current literature supports mandatory exploration or selective management. Computed tomographic (CT) scanning provides high-resolution images that are used for trauma in other body regions. The purpose of this study is to prospectively evaluate the utility of CT scanning in the evaluation of zone II penetrating neck injuries. METHODS: From July 1998 to November 1999, 14 stable patients were studied who sustained zone II penetrating neck injuries. All patients had a physical examination, infusion CT scan of the neck, and an operative exploration. Before surgery, the trauma surgeon evaluated the CT scan and interpreted it as demonstrating either "high" or "low" probability for significant injury. Surgical findings were compared with the surgeon's preoperative interpretation of the CT scan. RESULTS: Three of 14 patients had five significant injuries. All these patients had high probability of injury CT scans, with four of the five injuries specifically diagnosed by CT scan. Sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 91%, 75%, and 100% (p < 0.02), respectively. CONCLUSION: This small prospective study demonstrates that zone II penetrating neck injuries can be accurately evaluated by CT scan. In addition, the CT scan can be used for surgical decision making. This will eliminate the need for mandatory exploration and limit the role of angiography, esophagography, and endoscopy in zone II penetrating neck injuries. PMID- 11493792 TI - The routine use of sonography in penetrating torso injury is beneficial. AB - BACKGROUND: Torso sonography (focused assessment with sonography for trauma [FAST]) has been added to our protocols for the evaluation of penetrating torso injury. The purpose of this study was to evaluate our recent experience and determine whether the use of FAST is beneficial. METHODS: From January 1999 to January 2000, patients with penetrating torso injury and no clinical indication for surgery were evaluated by sonography with a selective use of other investigations. FAST consisted of sonographic views of the peritoneum and/or pericardium to determine the presence or absence of fluid. RESULTS: During the study period, there were 238 victims of penetrating injury assessed by our trauma service, and sonography was performed in 72 (30%) patients as per our protocols. There were 31 stab, 37 gunshot/shotgun and, and 4 puncture wounds. Thirty-eight patients had peritoneal views, 6 patients had pericardial views, and 28 patients had both pericardial and peritoneal views obtained. Thirteen of 66 patients had free fluid in the peritoneal cavity and 12 of the 13 patients had a therapeutic laparotomy. No peritoneal fluid was seen in 53 of 66 patients, of whom 6 had abdominal injuries, 5 requiring surgery for diaphragm or bowel injuries. The sensitivity of FAST alone for abdominal injury was 67%, specificity was 98%, positive predictive value was 92%, and negative predictive value was 89%. Pericardial fluid was seen in 3 of 34 patients; one had a heart wound and two had negative pericardial windows. All 31 patients without pericardial fluid recovered without surgery. CONCLUSION: The routine use of sonography in penetrating torso injury is beneficial. The detection of pericardial or peritoneal fluid is clinically useful. However, a negative FAST examination does not exclude abdominal injury, such as a diaphragm or hollow viscus wound, and further investigation or close follow-up is required. PMID- 11493793 TI - Stent-graft repair of acute traumatic thoracic aortic transection with intentional occlusion of the left subclavian artery: case report. PMID- 11493795 TI - Admission base deficit in pediatric trauma: a study using the National Trauma Data Bank. AB - BACKGROUND: The base deficit, an important indicator of physiologic derangement after severe injury in adults, has not been specifically examined in the pediatric trauma population. The purpose of this study was to assess the ability of the admission base deficit to predict injury severity and outcome in the pediatric trauma population. METHODS: The study group included all patients in the National Trauma Data Bank over a 2-year period aged 0 to 12 years with a base deficit (0 to -30 mEq/L) recorded from the emergency department. Age, presence of a severe closed head injury, and base deficit were analyzed with respect to mortality and other indicators of injury severity. RESULTS: A total of 515 patients constituted the study group. Base deficit less than -4 mEq/L (p < 0.001) and the presence of a closed head injury (odds ratio, 3.8; p < 0.05) were predictors of mortality. For the group, an admission base deficit of -8 mEq/L corresponded to a probability of mortality of 25%. Significant correlations were found between base deficit and emergency department systolic blood pressure, Injury Severity Score, and Revised Trauma Score. There was no relationship between age and mortality. CONCLUSION: In injured children, the admission base deficit reflects injury severity and predicts mortality. The probability of mortality increases precipitously in children with a base deficit less than -8 mEq/L, and should alert the clinician to the presence of potentially lethal injuries or uncompensated shock. PMID- 11493794 TI - Emotional considerations and attending involvement ameliorates organ donation in brain dead pediatric trauma victims. AB - PURPOSE: The purpose of this study was to ascertain a strategy for maximizing parental consent for organ donation in traumatically injured children suffering from brain death. Our hypothesis was that appropriate attending surgeon involvement and delay in evaluating children for brain death leads to an increased percentage of organ donors. METHODS: From January 1993 to August 1999, the records of all children who died in a Level I trauma center were evaluated. Those children suffering brain death that were suitable for organ donation were entered into the study. Cases were reviewed for patient demographics, time to entry into brain death protocol (measured from time of admission), time to parent notification about brain death (measured from time of admission), specific attending involved in the case (with level of involvement), and success of organ donation request. In all, 43 charts were reviewed. RESULTS: Of 43 deaths, 33 were deemed suitable for donation. Age of suitable donors ranged from 1 month to 18 years. In all, 11 attending physicians were involved in the care of these children. Overall, 20 of 33 were organ donors (60%). When the attending surgeon was involved, donation success for organ retrieval was 86%, whereas if the attending was not involved personally, the success rate dropped to 23% (p < 0.04). One senior pediatric surgeon obtained a success rate of 12 of 12 children. It was this surgeon's policy to not initiate brain death protocols in children immediately on entry into the emergency room, but rather to delay initiation until family could be gathered and spend time with the affected child in order that the family could recover from the initial shock of trauma (always at least overnight). When time to initiation of brain death protocol was examined, success was obtained when a delay of 15.5 hours was respected, versus 7.0 hours when donation was requested but denied (p < 0.03). CONCLUSION: These data indicate that attending involvement is important when parents of brain dead children are asked about organ retrieval (p < 0.04). Delay in initiating brain death protocols in order for family members to deal with the shock of the initial trauma appears to increase willingness to participate in organ donation. PMID- 11493796 TI - Introduction of a genitourinary trauma database for use as a multi-institutional urologic trauma registry. AB - BACKGROUND: Injury and/or work up of patients involved in genitourinary (GU) trauma often has a urologic component. By implementing a simple, versatile database to categorizing such injuries, we hoped to elucidate issues involved in the treatment of GU trauma. METHODS: The Genitourinary Trauma Database was created using commercially available software (Microsoft Access). It occupies 848 KB (kilobytes) of hard drive memory and is organized into five categories: Patient Demographics, History/Physical Examination/Etiology, Radiologic Evaluation, Operative Intervention, and Disposition and Complications. RESULTS: All GU trauma patients seen since 1995 have been entered. Thus far, the database has helped assess indications for bladder evaluation of blunt trauma and pelvic fracture and has allowed us to develop a protocol for performing retrograde computed tomographic cystogram in conjunction with abdominal/pelvic computed tomographic scan. CONCLUSION: Ease of creating and using this database demonstrate that it could serve as a multi-institutional GU trauma registry and prove invaluable in developing and refining future trauma protocols. PMID- 11493797 TI - Splenic injury in side impact motor vehicle collisions: effect of occupant restraints. AB - BACKGROUND: Side impact motor vehicle collisions (MVCs) are associated with higher morbidity and mortality compared with other types of MVCs. The stiffness of the lateral aspect of the vehicle and restraint use may play a role. The purpose of this study was to evaluate the role of restraint use, vehicle size, and compartment intrusion on the incidence of splenic injury in side impact MVCs. METHODS: The National Automotive Sampling System was used to identify drivers involved in side impact collisions for the years 1996 to 1998. The incidence of splenic injury in these collisions was compared according to restraint use, vehicle size, and magnitude of vehicle crush. Information on the perceived cause of splenic injuries sustained in the MVC was also obtained from National Automotive Sampling System investigator records. RESULTS: Overall, among drivers involved in side impact MVCs, restraint use was associated with a significantly reduced rate of mortality (odds ratio [OR], 0.40; p < 0.0001) and splenic injury (OR, 0.76; p < 0.0001). Restrained drivers of small vehicles (<2,500 lb), however, had a higher incidence of splenic injury in both minimal (lateral intrusion < 30 cm) (OR, 60.1; p < 0.0001) and severe (lateral intrusion > 30 cm) (OR, 4.0; p < 0.0001) magnitudes of vehicle crush on the driver's side of the vehicle. For both midsize (2,500-3,000 lb) and large (>3,000 lb) vehicles, restraint use was associated with a lower risk of splenic injury regardless of the magnitude of crush. In nearly all cases of splenic injury, the left vehicle interior was the source of injury. CONCLUSION: Overall, restraint use is associated with lower rates of splenic injury and mortality in side impacts. Despite this fact, restrained drivers of small vehicles have a higher risk of splenic injury after lateral impact MVCs when compared with unrestrained drivers. Evaluation of the combined role of restraint use, crash, and injury patterns may provide novel insight regarding vehicle safety design features. PMID- 11493798 TI - A statewide analysis of level I and II trauma centers for patients with major injuries. AB - BACKGROUND: This study examines statewide outcomes and resource use in Level I and II trauma centers for patients with major injuries. METHODS: This study analyzed trauma registry data on patients admitted to North Carolina Level I and II trauma centers from January 1995 to December 1996 with one of four major injuries: thoracic aortic disruption, liver injury, pelvic fracture, or pulmonary contusion. RESULTS: There were 59 thoracic aortic disruptions, 109 liver injuries, 153 pelvic fractures, and 962 pulmonary contusions identified among 26,030 admissions. Case fatality was not significantly different (Level I, 16.8%; Level II, 14.9%). Hospital charges were significantly higher in Level I centers (Level I, $47,366; Level II, $35,490), but this difference was confined to transferred patients. Controlling for Revised Trauma Score, Injury Severity Score, age, gender, and race, multivariable regression confirmed findings regarding hospital charges, and multiple logistic regression confirmed findings regarding case fatality. CONCLUSION: Case fatality was similar in Level I and Level II trauma centers in North Carolina, and hospital charges were comparable in patients with comparable injuries not requiring transfer. This suggests that patients with major injuries may be optimally cared for in both Level I and Level II trauma centers. PMID- 11493799 TI - Antibiotic prophylaxis for group A streptococcal burn wound infection is not necessary. AB - BACKGROUND: Historically, group A beta-hemolytic streptococci (GAS) burn wound infection has been a major source of morbidity and mortality in burn patients and has prompted the prophylactic administration of antibiotics to children with burns. Wound monitoring, surveillance cultures, and early excision of deep wounds may have changed this. Our objective in this project was to determine the efficacy of routine antibiotic prophylaxis in the era of early excision and closure of deep burn wounds. METHODS: Two cohorts of burned children were compared: all children admitted during calendar years 1992 through 1994 (group 1) and during calendar years 1995 through 1997 (group 2). All group 1 children received routine GAS antibiotic prophylaxis. Only those group 2 children with documented positive admission or surveillance cultures for GAS were treated. RESULTS: There were 511 children in group 1 and 406 children in group 2. They were well matched for age (4.7 +/- 0.21 years vs. 5.3 +/- 0.26 years, p = 0.06) and burn size (11.0% +/- 0.7% vs. 12.4% +/- 0.8%, p = 0.18). GAS species were recovered at admission or during hospitalization from 11 (2.6%) of group 1 children and 18 (4.4%) of group 2 children (p = 0.05), indicating a marginally higher rate of carriage in group 2. Nevertheless, in group 1 there were three (0.6%) who developed GAS wound infection and in group 2 there were four (0.98%, p = 0.71). The incidence of GAS infection in those patients with positive admission cultures was three (27%) of group 1 and four (22%) of group 2. No child developed fulminant GAS infection. CONCLUSION: Routine antibiotic prophylaxis of burn wounds in children in not effective in further reducing a low baseline incidence of GAS wound infection if admission screening by culture is used to identify those children who carry the organism and early excision of deep burns is practiced. PMID- 11493800 TI - Mesenteric and renal oxygen transport during hemorrhage and reperfusion: evaluation of optimal goals for resuscitation. AB - BACKGROUND: Changes in flow to the gut and the kidney during hemorrhage and resuscitation contribute to organ dysfunction and outcome. We evaluated regional and splanchnic oxygen (O2) flow distribution and calculated oxygen supply distribution during hemorrhage and reperfusion and compared them with global measures. METHODS: Seven anesthetized pigs were instrumented to evaluate global hemodynamics, visceral blood flow, and oxygen transport. Tonometric pH probes were positioned in the stomach and jejunum. Animals were bled to 45 mm Hg for 1 hour. Crystalloids and blood were infused during the following 2 hours to normalize blood pressure, heart rate, urine output, and hemo- globin. RESULTS: During hemorrhage, mesenteric flow and O2 consumption were significantly decreased, whereas systemic consumption remained normal. Renal flow was reduced, but renal O2 consumption remained normal. After resuscitation, despite normal hemodynamics, neither systemic, mesenteric, nor renal O2 delivery returned to baseline. Lactate remained significantly increased. Arterial pH, base excess, and gastric and jejunal pH were all decreased. CONCLUSION: During hemorrhage, the gut is more prone than other regions to O2 consumption supply dependency. After resuscitation, standard clinical parameters do not detect residual O2 debt. Lactate, arterial pH, base excess, and intramucosal gut pH are all markers of residual tissue hypoperfusion. PMID- 11493801 TI - Effect of acute anemia on the healing of intestinal anastomoses in the rabbit. AB - BACKGROUND: Considering the potential deleterious effects of transfusion, the effect of severe, acute anemia on small intestine (SI) and large intestine (LI) anastomoses was evaluated. METHODS: Thirty-six rabbits underwent segmental resection of the midileum and distal colon. Half underwent preanastomotic removal of 30% of their blood volume with crystalloid resuscitation. At 1 or 2 weeks, the anastomotic bursting pressure (ABP) and histologic grade of white blood cell influx, blood vessel ingrowth, fibroblast proliferation, and collagen deposition were assessed. RESULTS: SI ABP was decreased in anemic rabbits at 2 weeks (190 +/ 6 mm Hg vs. 257 +/- 17 mm Hg). LI ABP was decreased at 1 week in anemic rabbits. In the test rabbits, histologic parameters were altered in both the SI and LI with decreased white blood cell infiltration, attenuated blood vessel ingrowth, and decreased collagen content at both 1 and 2 weeks. Decreased SI fibroblast proliferation was present at 1 week. CONCLUSION: Acute anemia in the rabbit undergoing intestinal resection decreases the SI ABP at 2 weeks and alters histologic parameters of wound healing in both the SI and LI. PMID- 11493802 TI - Development and implementation of a clinical pathway for severe traumatic brain injury. AB - BACKGROUND: Clinical pathways (CPs) have been shown to be beneficial in optimizing patient care and resource use. METHODS: A multidisciplinary CP for the treatment of severe traumatic brain injury (Glasgow Coma Scale score of 3-7) was developed. Data from these patients (group I) were collected prospectively and compared with a retrospective database (group II). RESULTS: There were a total of 119 patients managed in conjunction with the CP and 43 patients in the control group. No statistical differences were found between the groups in age, Glasgow Coma Scale score at 24 hours, or Injury Severity Scores. There was a significant decrease in the length of hospital stay, intensive care unit stay, and length of ventilator support in the study group (group I: 22.5, 16.8, and 11.5 days, respectively; group II: 31.0, 21.2, and 14.4 days, respectively; p < 0.03). CONCLUSION: The use of this CP helped to standardize and improve patient care with fewer complications and a potential cost savings of approximately $14,000 per patient. PMID- 11493803 TI - Endoluminal stent-graft repair of a thoracic aortic transection in a trauma patient with multiple injuries: case report. PMID- 11493804 TI - Acetabular fractures with associated vascular injury: a report of two cases. PMID- 11493805 TI - Surgical complications of Ehlers-Danlos syndrome type IV: case report and review of the literature. PMID- 11493806 TI - Traumatic pneumobilia: case report. PMID- 11493807 TI - Acute colonic necrosis associated with sodium polystyrene sulfonate (Kayexalate) enemas in a critically ill patient: case report and review of the literature. PMID- 11493808 TI - Complete evisceration of the small intestine through a perianal wound as a result of suction at a wading pool. PMID- 11493809 TI - Traumatic and iatrogenic Horner syndrome: case reports and review of the literature. PMID- 11493810 TI - Unusual thoracoabdominal injury by fishing harpoon. PMID- 11493811 TI - Adhesion molecules and hemorrhagic shock. PMID- 11493812 TI - Tetanus: a new threat. PMID- 11493813 TI - Pulmonary parenchymal and bronchial arterial injuries secondary to blunt trauma. PMID- 11493814 TI - The problems with positive end expiratory pressure (PEEP) in association with abdominal compartment syndrome (ACS). PMID- 11493815 TI - The use of a survey to quantify the community's perceptions about neurosurgeons' involvement in trauma. PMID- 11493816 TI - The role of methylprednisolone in acute spinal cord injuries. PMID- 11493817 TI - The validity of the use of ultrasonography in the diagnosis of spontaneous and traumatic pneumothorax. PMID- 11493819 TI - Practitioners of pediatric emergency medicine: a 5-year longitudinal study. AB - STUDY OBJECTIVE: To describe the evolution of the responsibilities, goals and expectations of sub-Board-certified practitioners of pediatric emergency medicine (PEM) over a 5-year period. METHODS: This was a prospective, cohort study. A questionnaire was mailed in January 1994 to all physicians sub-Board-certified in PEM by either the American Board of Pediatrics or the American Board of Emergency Medicine. It included questions about the type of work the physicians did and expectations for the future. This group of physicians was surveyed again in January 1999. The primary outcome measures were changes in the physicians' goals and expectations for the future. Table. RESULTS: Questionnaires were mailed to 232 PEM sub-Board-certified physicians in January 1994. By June 1994, 183 of the 232 responded to the survey. Follow-up questionnaires were mailed to the cohort of 183 physicians in January 1999. By June 1999, 170 of the 183 (93%) had replied. The table summarizes results. In 1994, the most commonly listed career goals were to increase research productivity (52%) and develop excellent teaching skills (35%). In 1999, the most commonly listed goals were to improve hours/lifestyle (61%) and increase administrative work (33%). CONCLUSION: The priorities of this cohort of PEM sub-Board-certified physicians have changed as the physicians grow older. Lifestyle issues must be taken into consideration to ensure longevity in the subspecialty. PMID- 11493818 TI - Safety and efficacy of rocuronium for controlled intubation with paralytics in the pediatric emergency department. AB - BACKGROUND: Controlled intubation in the pediatric emergency department (ED) requires a paralytic agent that is safe, efficacious, and of rapid onset. The safety of succinylcholine has been challenged, leading some clinicians to use vecuronium as an alternative. Rocuronium's onset is similar to that of succinylcholine. OBJECTIVE: To evaluate the safety and efficacy of rocuronium for controlled intubation with paralysis (CIP) in the pediatric ED. METHODS: A retrospective, observational study reviewed the records of patients less than 15 years of age, who received controlled intubation with paralytics at two Dallas EDs. The patients received either vecuronium or rocuronium. RESULTS: The study included 84 patients (vecuronium 19, rocuronium 65). Complications were similar between the two groups. Rocuronium had a shorter time from administration to intubation when compared to vecuronium (P < 0.05). CONCLUSION: Rocuronium is as safe and efficacious as vecuronium for CIP in the pediatric ED. PMID- 11493820 TI - Do published guidelines predict pneumonia in children presenting to an urban ED? AB - OBJECTIVES: In 1997, a Canadian task force published evidence-based guidelines for diagnosing pediatric pneumonia, concluding that the absence of each of four signs (ie, respiratory distress, tachypnea, crackles, and decreased breath sounds) accurately excludes pneumonia. The study was performed to evaluate the accuracy of these guidelines in predicting pneumonia in young children. METHODS: This was an observational study conducted over a 4-month period at an urban emergency department with 80,000 annual visits, approximately 20% of which were children < or =5 years old. Consecutive children < or =5 years old who underwent chest radiography were enrolled. Prior to ordering radiographs, treating physicians were required to enter specific patient signs and symptoms into a computerized database. World Health Organization criteria were used to define tachypnea. Sensitivity, specificity, and predictive values of the task force guidelines in predicting pneumonia were calculated. RESULTS: Three hundred twenty nine children, including 67 (20%) with pneumonia, were enrolled. Guidelines were 45% sensitive (95% confidence interval (CI) = 33-58) and 66% specific (95% CI = 60-72) for diagnosing pneumonia. Positive and negative predictive values were 25% (95% CI = 18-34) and 82% (95% CI = 77-87), respectively. CONCLUSION: Previously published evidence-based guidelines for excluding pediatric pneumonia were found unreliable in this study. PMID- 11493822 TI - Ketamine sedation for pediatric critical care procedures. AB - OBJECTIVES: To describe our experience using ketamine sedation to facilitate pediatric critical care procedures, and to document the safety profile of ketamine in this setting. DESIGN: Retrospective consecutive case series. SETTING: Pediatric intensive care unit of a tertiary children's hospital. PATIENTS: Children receiving ketamine for procedural sedation over a 5-year period. INTERVENTIONS: We reviewed patient records to determine indication, dosing, adverse events, inadequate sedation, and recovery time for each sedation. OUTCOME MEASURES: Descriptive features of sedation including adverse events. RESULTS: During the study period, children in our pediatric intensive care unit received ketamine at total of 442 times to facilitate a wide variety of critical care procedures, most commonly central line placement, esophagogastroduodenoscopy, and wound debridement. Most study children had substantial underlying illness (ASA > or = 3 in 88%; ASA > or = 4 in 39%). Inadequate sedation was noted in only nine (2%) procedures. Adverse effects included transient laryngospasm (n = 9), transient partial airway obstruction (n = 5), apnea with bradycardia (n = 1), emesis during the procedure (n = 2), emesis during recovery (n = 9), mild recovery agitation (n = 10), moderate-to-severe recovery agitation (n = 1), and excessive salivation (n = 4). There were no adverse outcomes attributable to ketamine. CONCLUSION: Pediatric intensivists skilled in ketamine administration can safely and effectively administer this drug to facilitate critical care procedures. Despite the ill nature of our patient sample, adverse effects were uncommon. PMID- 11493823 TI - Home antipyretic use in children brought to the emergency department. AB - OBJECTIVE: To determine how often children with a complaint of fever receive antipyretics at home and if any demographic factors are associated with correct dosing. METHODS: A prospective, descriptive study of children 3 to 36 months old presenting with complaint of fever was conducted. Caregivers were questioned about demographics and antipyretic given. The ability of demographics to predict proper dosing was tested first individually and then with a regression model. The effect of proper home dosing on presence of fever and height of fever was also analyzed. A total of 138 children were analyzed. RESULTS: Of the 118 (86%) who received antipyretics at home, only 47% had been given a proper dose. No demographic variable predicted proper dosing. CONCLUSION: Reported antipyretic dose at home did not predict presence of fever or height of temperature measured in the emergency department. PMID- 11493824 TI - Lenticular lesions: not always an epidural hematoma. AB - Ewing's sarcoma is a tumor of the bone, which occurs most often in the diaphysis of long and flat bone. The most common sites of metastasis are the lungs and bones. Less frequently, the primary site is an intracranial or pelvic lesion (either as a soft tissue or a bone lesion). We report a case of a 16-year-old female with an extraosseous intracranial lesion, who presented with a history of minor trauma, unilateral facial swelling, and head pain. Though head computed tomography scan showed a lesion consistent with an epidural hematoma, further exploration revealed Ewing's sarcoma. PMID- 11493825 TI - Retropharyngeal abscess in a toxic-appearing infant. PMID- 11493826 TI - Vertebral fractures as initial signs for acute lymphoblastic leukemia. AB - Common presenting symptoms of acute lymphoblastic leukemia in children are well known and include pallor, fatigue, and loss of appetite. Limb pain is sometimes described and can be misleading. We describe two recent cases seen in our emergency department, where vertebral fractures, a much rarer finding, were the only presenting symptoms that led to the diagnosis. One case had been thoroughly evaluated only 5 weeks prior to the diagnosis and included magnetic resonance imaging. The second patient was rapidly referred to our center with a history of acute lumbar pain. Emergency physicians caring for children must be aware of this rare type of presentation of leukemia. PMID- 11493827 TI - Spontaneous pneumomediastinum in a shallow-water--diving child. PMID- 11493828 TI - Traumatic appendicitis: fact or fiction? PMID- 11493829 TI - A football player with thigh pain. PMID- 11493830 TI - Delayed presentations of congenital diaphragmatic hernia. AB - The delayed presentation of a congenital diaphragmatic hernia may include symptoms of acute respiratory distress or gastrointestinal obstruction. We review the presentation and management of the different types of diaphragmatic hernias. In addition, we present a case of twins with a left-sided congenital diaphragmatic hernia through the foramen of Morgagni and an associated malrotation of the gastrointestinal tract. PMID- 11493831 TI - Rapid strep tests in the emergency department: an evidence-based approach. PMID- 11493832 TI - Sometimes it's not so clear: altered mental status and transport. PMID- 11493833 TI - Telemedicine in a box: overcoming complexity and high-cost telemedicine barriers using self-contained videoconferencing units. AB - This report describes our experience using low-and moderate-cost videoconferencing systems for telemedicine. After determining that low-cost systems using a standard personal computer and personal computer camera were unsatisfactory, a demonstration project was carried out using a $4000 self contained videoconferencing unit (telemedicine in a box), using eight simulated telemedicine consultation scenarios. The quality of the videoconferencing was good for all eight scenarios. All eight consultation simulations demonstrated different ways of improving patient care. Two of the major problems hindering the broad proliferation of telemedicine (high cost and high complexity) are solved by the telemedicine in a box concept. Focussing on the telemedicine in a box concept when planning a telemedicine system will improve its feasibility in the real world of health care. PMID- 11493834 TI - Practice styles. PMID- 11493835 TI - Case records of the Children's Hospital of Michigan: a 15-year-old with vomiting and weight loss. PMID- 11493836 TI - Mild knee injury with prolonged problems. PMID- 11493837 TI - Pediatric emergency medicine: legal briefs. PMID- 11493838 TI - Recognizing suicide risk in a pediatric emergency department: a change in nursing care. PMID- 11493839 TI - Pumpkin seed aspiration into the middle of the trachea in a wheezy infant unresponsive to bronchodilators. PMID- 11493840 TI - Applications of heliox in the treatment of status asthmaticus. PMID- 11493841 TI - Retinal hemorrhages in infants with hyponatremic seizures. PMID- 11493842 TI - Gamma-hydroxybutyrate, gamma-butyrolactone, and 1,4-butanediol. PMID- 11493843 TI - A randomized controlled trial to prevent patient lift and transfer injuries of health care workers. AB - STUDY DESIGN: Randomized controlled trial (RCT). OBJECTIVES: To compare the effectiveness of training and equipment to reduce musculoskeletal injuries, increase comfort, and reduce physical demands on staff performing patient lifts and transfers at a large acute care hospital. SUMMARY OF BACKGROUND DATA: Back injury to nursing staff during patient handling tasks is a major issue in health care. The value of mechanical assistive devices in reducing injuries to these workers is unclear. METHODS: This three-armed RCT consisted of a "control arm," a "safe lifting" arm, and a "no strenuous lifting" arm. A medical, surgical, and rehabilitation ward were each randomly assigned to each arm. Both intervention arms received intensive training in back care, patient assessment, and handling techniques. Hence, the "safe lifting" arm used improved patient handling techniques using manual equipment, whereas the "no strenuous lifting" arm aimed to eliminate manual patient handling through use of additional mechanical and other assistive equipment. RESULTS: Frequency of manual patient handling tasks was significantly decreased on the "no strenuous lifting" arm. Self-perceived work fatigue, back and shoulder pain, safety, and frequency and intensity of physical discomfort associated with patient handling tasks were improved on both intervention arms, but staff on the mechanical equipment arm showed greater improvements. Musculoskeletal injury rates were not significantly altered. CONCLUSIONS: The "no strenuous lifting" program, which combined training with assured availability of mechanical and other assistive patient handling equipment, most effectively improved comfort with patient handling, decreased staff fatigue, and decreased physical demands. The fact that injury rates were not statistically significantly reduced may reflect the less sensitive nature of this indicator compared with the subjective indicators. PMID- 11493844 TI - Intervertebral disc cells exhibit differences in gene expression in alginate and monolayer culture. AB - STUDY DESIGN: The mRNA levels of aggrecan and collagen were quantified in intervertebral disc cells cultured under three conditions: primary alginate culture, monolayer culture, and re-encapsulation in alginate after monolayer culture. OBJECTIVES: To compare the phenotype of intervertebral disc cells under different culture conditions and to investigate the reversibility of cell phenotype after re-encapsulation in alginate after monolayer culture. SUMMARY OF BACKGROUND DATA: The intervertebral disc contains heterogeneous populations of cells that vary with anatomic region. These cells possess significant differences in phenotype that can be preserved in vitro, although the effect of culture conditions on the phenotype of these cells is poorly understood. METHODS: The intervertebral disc cells of 4-5-month-old pigs were isolated enzymatically from three anatomic zones: anulus fibrosus (AF), transition zone (TZ), and nucleus pulposus (NP). Gene expression levels of aggrecan and collagen Types I and II were measured using a quantitative reverse transcriptase--polymerase chain reaction. RESULTS: Gene expression levels of anulus fibrosus and transition zone cells were shifted in monolayer compared with alginate, although the shift was partially reversed when re-encapsulated in alginate. However, NP cells appeared to be insensitive to culture conditions. Furthermore, characteristic patterns of gene expression among AF, TZ, and NP cells in primary alginate culture did not exist in monolayer culture, but they were also observed after re-encapsulation in alginate. CONCLUSION: The findings of this study suggest that anulus fibrosus and transition zone cells undergo a reversible shift in phenotype when cultured in monolayer compared with alginate. These differences suggest that the culture system exerts a strong influence on cell phenotype and may play a role in the response of these cells to biophysical and biochemical stimuli in vitro. PMID- 11493846 TI - Peak stresses observed in the posterior lateral anulus. AB - STUDY DESIGN: The stress distributions within cadaveric lumbar intervertebral discs were measured for a range of loading conditions. OBJECTIVES: To examine the distribution of stress across the area of the intervertebral disc and to compare regional variations in peak stress during compression loading with various flexion angles. SUMMARY OF BACKGROUND DATA: The rate of disc degeneration and the occurrence of low back disorders increase with higher mechanical loading of the spine. The largest peak stresses occur in the anulus. METHODS: Human lumbar L2- L3 and L4--L5 cadaver functional spinal units were obtained and tested. The distribution of disc stress was measured using a pressure probe with loads applied, pure compression and compression with 5 degrees of either flexion or extension. RESULTS: Stress profiles were recorded across the intervertebral disc at a compressive force of 1000 N and each of the three flexion-extension angles. The highest values (2.99 +/- 1.31 MPa) were measured during extension-compression lateral to the midline of the disc in the posterior anulus. The pressure in the nucleus was relatively unchanged by flexion angle remaining about 1.00 MPa for a 1000-N compression. CONCLUSIONS: Pressure measurements of the cadaveric nucleus have been used to validate models of lumbar spine loading and to evaluate the risk of low back injury and disc herniation. Previous observations limited to midsagittal measurements of the nucleus did not identify the regions of highest stress. The highest values observed here within the posterolateral anulus correspond to common sites of disc degeneration and herniation. PMID- 11493847 TI - The Torg--Pavlov ratio in cervical spondylotic myelopathy: a comparative study between patients with cervical spondylotic myelopathy and a nonspondylotic, nonmyelopathic population. AB - STUDY DESIGN: A radiologic study to compare the Torg--Pavlov ratios between patients with cervical spondylotic myelopathy and a nonspondylotic, nonmyelopathic population. OBJECTIVES: To determine and compare the Torg--Pavlov ratios between the two groups of patients. SUMMARY OF BACKGROUND DATA: Patients with congenital cervical spinal canal stenosis are more likely to develop cervical spondylotic myelopathy. The Torg--Pavlov ratio eliminates errors related to magnification, a problem with determination of spinal canal stenosis from direct measurements of plain cervical spine radiographs. There has only been one other study that directly compares the Torg--Pavlov ratio between patients with cervical spondylotic myelopathy and a normal control population. METHODS: The preoperative plain lateral cervical spine radiographs of 28 patients with cervical spondylotic myelopathy requiring surgical decompression were compared with radiographs of 88 nonspondylotic, nonmyelopathic patients. The Torg--Pavlov ratio was computed for each level from C3 to C7. RESULTS: The study showed that the Torg--Pavlov ratio is significantly smaller (P < 0.001) in myelopathic patients (mean 0.72 +/- 0.08) compared with the control patients (mean 0.95 +/- 0.14). This was so when individual levels and the mean values were compared. Age was also found to be a significant factor (P = 0.002), although lesser in magnitude when compared with the Torg--Pavlov ratio (P = 0.0001). CONCLUSIONS: The Torg--Pavlov ratio is significantly lower in patients with cervical spondylotic myelopathy compared with a nonspondylotic, nonmyelopathic population. It could possibly be used to predict the likelihood of developing cervical spondylotic myelopathy. PMID- 11493848 TI - Fibrocartilage in the transverse ligament of the human atlas. AB - STUDY DESIGN: Immunohistochemical investigation. OBJECTIVE: To determine whether molecules typical of articular cartilage are present in the transverse ligament and whether the ligament may be a target for an autoimmune response in rheumatoid arthritis. SUMMARY OF BACKGROUND DATA: In chronic rheumatoid arthritis there is often a marked instability of the atlantoaxial complex, and the transverse ligament can show degenerative changes that compromise its mechanical function. In some rheumatoid patients there can be an autoimmune response to cartilage link protein, aggrecan, and Type II collagen. METHODS: Transverse ligaments were removed from 13 cadavers and fixed in 90% methanol. Cryosections were immunolabeled with antibodies against proteoglycans (aggrecan, link protein, and versican), glycosaminoglycans (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, and keratan sulfate), and collagens (Types I, II, III, and VI). RESULTS: Labeling for aggrecan and link protein was characteristic of the fibrocartilages, but versican was only detected in the fibrous regions. Equally, Types I, III, and VI collagens and keratan, dermatan, and chondroitin-4-sulfates were found throughout the ligament, but labeling for Type II collagen and chondroitin-6-sulfate was restricted to the fibrocartilages. CONCLUSION: The presence of molecules typical of articular cartilage (aggrecan, link protein, and Type II collagen) in the transverse ligament explains why it can be a target for destruction in rheumatoid arthritis and also suggests that it is subject to constant compression against the dens rather than only at the extremes of movement. PMID- 11493849 TI - Evaluation of various evoked potential techniques for spinal cord monitoring during scoliosis surgery. AB - STUDY DESIGN: This prospective study compared the outcomes of different evoked potential (EP) techniques for intraoperative spinal cord monitoring. OBJECTIVES: To evaluate the reliability of different EP techniques administered during scoliosis surgery. SUMMARY OF BACKGROUND DATA: A number of different methods of intraoperative spinal cord monitoring are available. Because each has its own advantages and limitations, multimodal spinal cord monitoring has been proposed to improve monitoring reliability. MATERIALS AND METHODS: Cortical somatosensory evoked potential (CSEP), cortical motor-evoked potential (CMEP), spinal somatosensory-evoked potential (SSEP), and spinal cord-evoked potential (SCEP) were applied simultaneously to 30 patients undergoing surgical correction for spinal deformity. The presence of the EP waveforms and their reproducibilities over separate tests were compared. In addition, the monitoring outcomes were evaluated with the clinical results. RESULTS: Of the 30 patients, CSEP waveforms were successfully recorded in 28 cases (93%), SCEP in 25 cases (83%), CMEP in 24 cases (80%), and SSEP in 21 cases (70%). Latencies of each EP technique showed no significant variability. However, amplitudes showed significant differences between different techniques. SCEP and CMEP showed clearer waveforms of greater amplitude that could be detected faster than CSEP and SSEP waveforms. SCEP and SSEP waveforms were more easily influenced by the surgical procedure. CONCLUSION: CSEP and CMEP are recommended for routine monitoring, so that both ascending and descending tracts are monitored. If adequate signals for either of these proposed monitoring methods cannot be easily obtained, SSEP can substitute for CSEP, whereas SCEP can substitute for CMEP. PMID- 11493850 TI - Changes in surface and radiographic deformity after Universal Spine System for right thoracic adolescent idiopathic scoliosis: is rib-hump reassertion a mechanical problem of the thoracic cage rather than an effect of relative anterior spinal overgrowth? AB - STUDY DESIGN: Analysis of preoperative, 8-week, 1-year, and 2-year data from patients with right thoracic adolescent idiopathic scoliosis treated by posterior Universal Spine System (Stratec Medical, Oberdorf, Switzerland). OBJECTIVE: Report 2-year results and the association between back surface and radiographic assessments. SUMMARY OF BACKGROUND DATA: Few longitudinal studies have related surface and radiographic data in the follow-up of surgical patients. METHODS: Of 34 patients with right thoracic adolescent idiopathic scoliosis having posterior Universal Spine System instrumentation, 27 had complete prospective back surface and radiographic appraisal. RESULTS: Cobb angle corrected from 58 degrees to 34 degrees (41%), apical vertebral rotation from 26 degrees to 20 degrees (23%), apical vertebral translation from 4.5 to 2.4 cm (47%), and maximum angle of trunk inclination from 17 degrees to 13 degrees (22%) (preoperative to 2 years). Rib hump reassertion occurred between 8 weeks and 1 year, regardless of age, and correlated with changes in vertebral translation (for 10 vertebral levels corresponding to 10 back surface levels between C7 and S1, P = 0.001 MANOVA). Preoperative frontal tilt of L1 with concave fifth rib-spinal angle predicted the percentage correction of maximum angle of trunk inclination, and the concave ninth rib-spinal angle predicted reassertion of maximum angle of trunk inclination. CONCLUSIONS: Almost half of initial back surface correction is lost by 2 years. Segmental vertebral translation measurements most strongly correlate with segmental angle of trunk inclination measurements during follow-up. Rib-hump reassertion is best explained by unwinding of the thoracic cage tensioned by surgery rather than through relative anterior spinal overgrowth. Spine and thoracic cage factors determine rib-hump correction, so surgical disruption of the latter by costoplasty may prevent rib-hump reassertion. Results of scoliosis surgery should include surface data. PMID- 11493851 TI - The association between physical workload and low back pain clouded by the "healthy worker" effect: population-based cross-sectional and 5-year prospective questionnaire study. AB - STUDY DESIGN: A population-based cross-sectional and 5-year prospective questionnaire study. OBJECTIVE: To investigate self-reported physical workload as a risk factor for low back pain. SUMMARY OF BACKGROUND DATA: Both physical and psychosocial workplace factors are considered risk factors for low back pain. However, today no consensus has been reached regarding the exact role of these factors in the genesis of low back pain. METHODS: Questionnaire data were collected at baseline for 1397 (and after 5 years for 1163) men and women aged 31 -50 years at baseline. Low back pain ("any low back pain within the past year," "low back pain < or = 30 days in total during the past year," "low back pain > 30 days in total during the past year") was analyzed in relation to physical workload (sedentary, light physical, and heavy physical work) using logistic regression and controlling for age, gender, and social group. The proportions of workers changing between the workload groups over the 5-year period were analyzed in relation to low back pain status. RESULTS: At baseline no statistically significant differences in low back pain outcomes were found for workers exposed to sedentary, light physical, or heavy physical work. This was true for all age, gender, and social groups. At follow-up there was a statistically significant dose-response association between any low back pain and longstanding low back pain within the past year and increasing physical workload at baseline also after controlling for age, gender, and social group. Subjects with heavy physical workload at baseline changed statistically significantly more often to sedentary work if they experienced low back pain for more than 30 days out of the past year. CONCLUSIONS: Having a sedentary job might have a protective or neutral effect in relation to low back pain, whereas having a heavy physical job constitutes a significant risk factor. Because of migration between exposure groups (the "healthy-worker" effect), longitudinal studies are necessary for investigating the associations between physical workload and low back pain. PMID- 11493853 TI - The effect of wearing a back belt on spine kinematics during asymmetric lifting of large and small boxes. AB - STUDY DESIGN: A crossover design was used to evaluate kinematic measurements collected with an infrared-based motion measurement system. OBJECTIVES: To evaluate belt effects on spine kinematics during asymmetric lifting of large and small boxes and to test for carryover effects between trials from belts. SUMMARY OF BACKGROUND DATA: Conflicting evidence in the literature exists regarding whether belts are beneficial or detrimental to manual material handlers. Studies have not examined belt effects when lifting different sized boxes, nor carryover effects from belts. METHODS: Twenty-eight subjects with manual-handling experience (17 male and 11 female) were randomly assigned to lift either a large or small box (weighing 9.4 kg), from a sagittally symmetric origin at pallet height to a 79 cm height, 60 degrees to the right. Spine flexion, lateral bending and twisting, hip and knee flexion, and angular velocity measurements of the torso with respect to the pelvis were collected for each of three lifting periods, 50 lifts each at 3 lifts per minute, with 18-minute breaks between periods. RESULTS: Belts significantly reduced maximum spine flexion, spine flexion and extension angular velocities, and torso left lateral bending angular velocity, and increased hip and knee flexion, regardless of box size. When lifting large boxes, belts significantly reduced torso right lateral bending and torso left twisting. No significant differential carryover effects were detected from belts. CONCLUSIONS: Subjects with belts lifted more slowly and used more of a squat-lift technique, regardless of box size. Belts reduced more torso motions while lifting large boxes. PMID- 11493854 TI - Within-subject variability in low back load in a repetitively performed, mildly constrained lifting task. AB - STUDY DESIGN: A repeated-measures in vivo experiment. OBJECTIVE: To describe within-subject variability of spinal compression in repetitive lifting. SUMMARY OF BACKGROUND DATA: Epidemiology and failure mechanics suggest that peak loads may be more predictive of injury than average loads. Nevertheless, biomechanical studies usually focus on the latter. METHODS: Ten healthy males performed 360 lifts in 1 hour of a 45-L crate, weighted with a stable 10-kg mass on 1 day and with an unstable mass (10 kg of water) on another day. The maximum compression force in each lift was estimated, using a simple inverse dynamics model and a single equivalent muscle model. RESULTS: The individual distributions of maximum compression force were slightly skewed to the right (average skewness 0.67). Median and 95th percentile values were used to characterize the distribution. The median (50th percentile) compression ranged from 3375 to 6125 N, and from 3632 to 6298 N in the stable and unstable load conditions, respectively. The within subjects peak (95th percentile) compression forces were from 405 to 1767 N and from 526 to 2216 N, respectively, higher than the median values. The peak values differed significantly between conditions, whereas the difference in medians did not reach significance. Only a limited trendwise (fatigue-related) variance could be demonstrated. CONCLUSION: Peak spinal compression by far exceeds median compression in repetitive lifting and can be affected by task conditions independently from the median. Therefore, the variability of spinal loads needs to be taken into consideration when analyzing and redesigning tasks that can cause spinal injuries. PMID- 11493855 TI - A novel technique for surgical resection of spinal meningioma. AB - STUDY DESIGN: A technical note. OBJECTIVES: To describe and discuss a novel technique for surgical resection of spinal meningioma. SUMMARY OF BACKGROUND DATA: With conventional methods for surgical resection of isolated intradural spinal meningioma, there are two ways of dealing with the dural attachment of the tumor. One is complete resection of the involved dura together with the tumor, and the other is coagulation only of the tumor base of the dura. In the case of the novel technique herein described, the dura mater is preserved in a new manner. METHODS: In the surgical procedure a small incision is made in the surface of the dura mater after the conventional laminectomy. The spinal dura can be easily divided into two layers, comprising inner and outer layers. The outer layer is stripped away from the inner layer surrounding the tumor base. The tumor is then resected together with the inner layer alone outside the arachnoid membrane, and finally the outer layer is simply closed. RESULTS: This method has been applied to three cases. The preserved outer layer of the dura mater did not demonstrate the existence of tumor cells histologically. Neither complications nor tumor recurrence have been experienced. CONCLUSION: The authors introduced a novel technique for surgical resection of isolated intradural spinal meningioma. Using this simple procedure the outer part of the dura mater, which is not involved by the tumor, can be preserved and complicated dural reconstruction is not necessary. Furthermore, there is less risk of postoperative cerebrospinal fluid fistulas than when the dura is completely resected together with the tumor. However, long-term observation as a result of the possibility of local recurrence is strongly recommended. PMID- 11493856 TI - Subarachnoid--pleural fistulae--management with a transdiaphragmatic pedicled greater omental flap: report of two cases. AB - STUDY DESIGN: This report describes the treatment of chronic subarachnoid- pleural fistulae using a pedicled greater omentum transfer flap. OBJECTIVE: To describe a new technique for the management of chronic subarachnoid--pleural fistulae resulting from thoracic dural tears. SUMMARY OF BACKGROUND DATA: Thoracic dural tears with leakage of cerebral spinal fluid into the pleural space can occur after thoracic spine surgery. The treatment of chronic subarachnoid- pleural fistulae using an omental flap, however, has not been reported. METHODS: The clinical, radiographic, and surgical details of two cases are described. RESULTS: Pedicled greater omentum transferred to the thoracic spine was an effective method in the treatment of intractable thoracic dural tears for two patients. CONCLUSION: These cases demonstrate that pedicled greater omentum transferred to the thoracic spine can be a safe and effective technique for the management of intractable thoracic dural tears and their complications. PMID- 11493857 TI - Oral extrusion of a screw after anterior cervical spine plating. AB - STUDY DESIGN: A case report of a 76-year-old woman who retched up a screw from a cervical spine locking plate 5 years after anterior cervical spine fusion. The literature relevant to this topic is reviewed. OBJECTIVES: To report the rare but potentially life-threatening complication of oral screw extrusion after anterior cervical spine plating, to review the relevant literature on the topic, and to discuss the clinical management of instrumentation failure in anterior cervical spine plating. SUMMARY OF BACKGROUND DATA: Anterior cervical spine fusion and stabilization is a well-established procedure. Complications include instrumentation failure, which can progress to extrusion through the gastrointestinal tract. Management is dependent on the severity and progression of clinical and radiologic signs and symptoms. Reoperation should be considered in certain cases. METHODS: A rare complication of anterior cervical spine plating in a 76-year-old woman 5 years after the initial operation is reported. The patient was assessed with serial physical examination and radiograph and one further follow-up 3 months after the first presentation. RESULTS: The patient was asymptomatic shortly after she retched up the screw, and at the 3-month follow-up was without evidence of progression of plate dislodgement. CONCLUSION: As reported, oral extrusion of cervical spine grafts or instrumentation is rare but potentially serious. Each case of instrumentation failure should be assessed individually to decide if conservative management is appropriate or if reoperation should be considered. PMID- 11493858 TI - Presacral schwannoma with purely cystic form. AB - STUDY DESIGN: Case report. OBJECTIVE: To point out that presacral schwannoma can display markedly cystic degeneration. SUMMARY OF BACKGROUND DATA: Benign schwannoma sometimes displays degenerative changes, such as cyst formation, calcification, hemorrhage, and hyalinization. Usually these degenerations are partially seen in the tumors. METHODS: Medical history, physical findings, imaging features, and histologic findings were reviewed in a case with a large presacral schwannoma. RESULTS: A 62-year-old woman presented with an 18-month history of right sciatic pain. Magnetic resonance images detected a large cystic, presacral tumor. Initially the tumor was misdiagnosed as a cystic ovarian tumor. Preoperative diagnostic workup led to the diagnosis of cystic schwannoma. Intralesional curettage achieved good pain relief. CONCLUSIONS: This tumor was unusual in its totally cystic appearance and resemblance to a cystic ovarian tumor or anterior sacral meningocele. Preoperative diagnosis is important to prevent major neurologic deficit at surgery. PMID- 11493859 TI - Salvage reconstruction with vascularized fibular strut graft fusion using posterior approach in the treatment of severe spondylolisthesis. AB - STUDY DESIGN: One case is reported in which a failed anterior fusion for Grade 4 spondylolisthesis was treated with a vascularized fibular strut graft using a posterior approach. OBJECTIVES: To demonstrate the applicability of this technique for salvage cases or patients with systemic conditions that may decrease the success of more standard techniques. SUMMARY OF BACKGROUND DATA: Surgical stabilization of spondylolisthesis through posterior approach with a fibular strut graft has been previously described. A vascularized strut graft can be used in the treatment of spondylolisthesis and may have applicability in those patients with underlying disease that may impair the use of more standard techniques or in salvage reconstruction. METHODS: With the patient under general anesthesia, through a posterior approach S1 and L4 were decompressed. The fibula with its vascularity intact was harvested and anastomosed with the superior gluteal artery and vein. The fibular strut was placed into the space formed by reaming between L5 and S1. Ilial autograft was used to augment the posterior fusion. After the procedure the patient was placed in a hip spica cast. RESULTS: At the 2-year follow-up the patient has incorporation of the graft, with no evidence of fracture and no significant progression of anterior slip. CONCLUSION: A vascularized fibular strut graft is a feasible alternative in the treatment of severe spondylolisthesis. No complications were encountered in the involved patient. Future application may include salvage reconstruction of failed arthrodesis or in individuals with systemic conditions that may impair graft incorporation using more standard techniques. PMID- 11493860 TI - Imaging corner: unknown case. PMID- 11493861 TI - Re: van Tulder et al, Exercise therapy for low back pain. Spine 2000;25(21):2784- 96. PMID- 11493862 TI - Re: Exercise therapy for low back pain. A systematic review within the framework of the Cochrane Collaboration Back review Group. Spine 2000;25:2784--96. PMID- 11493863 TI - Re: van Tulder et al, Exercise therapy for low back pain. Spine 2000;25:2784--96. PMID- 11493864 TI - Transpedicular wedge osteotomy for correction of thoracolumbar kyphosis in ankylosing spondylitis: experience with 78 patients. AB - STUDY DESIGN: This is a retrospective study of surgical correction of thoracolumbar kyphosis caused by ankylosing spondylitis. OBJECTIVE: To report the surgical results of thoracolumbar kyphosis deformity corrected with transpedicular wedge osteotomy performed by a single surgeon at a university hospital. SUMMARY OF BACKGROUND DATA: There has not been a large series in the literature reporting on results of the Thomasen-type closing wedge osteotomy for correction of kyphosis deformity secondary to ankylosing spondylitis, nor has two level osteotomy of this type in one patient ever been described. METHODS: From 1991 through 1998, 92 transpedicular wedge osteotomies were performed in 78 patients with ankylosing spondylitis for correction of fixed flexion deformity of the thoracolumbar spine. RESULTS: The mean amount of correction for each level of osteotomy was 34.5 degrees (range, 15 degrees -60 degrees ). The largest amount of overall correction for a single patient was 100 degrees. Most of the osteotomies (64 of 92) were done at L2 and L3. Fourteen patients with severe deformity required staged two-level osteotomy. Excellent and good results were obtained in 77 patients (98.7%) at the final follow-up. There was no mortality, nor were there any major neurological complications. CONCLUSIONS: Transpedicular wedge osteotomy can effectively and safely correct kyphotic deformity of the thoracolumbar spine caused by ankylosing spondylitis, regardless of rigidity of the spinal curves. Two-level osteotomy can provide sufficient correction for severe cases. PMID- 11493865 TI - Back muscle fatigability is associated with knee extensor inhibition in subjects with low back pain. AB - STUDY DESIGN: Cross-sectional study of 25 male golfers with chronic low back pain and 16 healthy controls of similar age. OBJECTIVES: To assess the association between functional capacity of the back extensors and the quadriceps muscles. SUMMARY OF BACKGROUND DATA: Chronic low back pain has been shown to lead to changes in muscle activation patterns of the abdominals and the gluteus maximus. The effect of chronic low back pain on lower limb function has not been investigated. METHODS: Back extensor endurance was assessed by a Biering-Sorensen test; surface EMG was measured bilaterally on the erector spinae at T12 and L4- L5. Muscle inhibition in the quadriceps was assessed by applying an electrical twitch to the maximally contracted muscle. The associations between holding time, decrease in EMG median frequency (i.e., the slope of the regression line on median frequency vs. time), and muscle inhibition were compared for study participants with chronic low back pain and controls. RESULTS: Mean back extensor holding times were 88 +/- 30 seconds for study participants with chronic low back pain and 92 +/- 17 seconds for controls. Both groups showed bilaterally similar decreases in EMG median frequency at L4--L5 and T12; however, the slopes were significantly steeper at L4--L5 than T12. Study participants with chronic low back pain with poor back endurance had significantly higher muscle inhibition compared with study participants with chronic low back pain with good back endurance, whereas such an association was not evident in healthy controls. CONCLUSIONS: In golfers with chronic low back pain reduced back endurance was associated with significant inhibition of the knee extensors, indicating that this muscle group cannot be activated to a full extent. These findings suggest a possible association between back extensor fatigability and knee extensor dysfunction in male golfers with chronic low back pain. PMID- 11493866 TI - Disc herniation-related back pain impairs feed-forward control of paraspinal muscles. AB - STUDY DESIGN: A comparative study of lumbar paraspinal muscle reflexes during sudden upper limb loading in healthy control subjects and patients with sciatica. OBJECTIVES: To assess reflex activation of paraspinal muscles during sudden upper limb loading. SUMMARY OF BACKGROUND DATA: Sudden upper limb loading and upper limb voluntary movements cause reflex activation of trunk muscles. A short latency response of approximately 50 msec of lumbar muscles has been observed before, but the reflexes have not been studied in patients with sciatica. METHODS: The paraspinal muscle responses for upper limb loading during unexpected and expected conditions were measured by surface EMG from 20 patients selected for an operation as a result of disc herniation-related chronic low back pain and 15 back-healthy controls. Pain, disability, and depression scores were recorded. RESULTS: Short latency response of paraspinal muscles for unexpected upper limb loading was similar in healthy controls and patients with sciatica in supported standing. During normal standing anticipation shortened the lumbar reflex latency in healthy controls but not among the patients. CONCLUSIONS: The results provide evidence for impaired feed-forward control of lumbar muscles in patients with sciatica. PMID- 11493867 TI - Changing curve pattern in infantile idiopathic scoliosis: family report with a follow-up of 15 years. AB - STUDY DESIGN: The authors monitored a father and his six children, all of whom had idiopathic scoliosis for more than 13 years. OBJECTIVE: To analyze the similarities and differences in curve pattern between the family members. SUMMARY OF BACKGROUND DATA: The etiology and pathogenesis of idiopathic scoliosis are not well understood. Genetic factors play an important role, as is shown by the high concordance in monozygotic twins. METHODS: Radiographs from all family members were analyzed. Location and direction of the curve, Cobb angle, location of the apex, and rib-vertebra angle difference were compared. RESULTS: A father and his six children had idiopathic scoliosis. The curves differed within the family and changed during follow-up. The father had a left convex thoracic scoliosis of 42 degrees. Only three of the children had a curve of similar severity, and only four children had a left convex thoracic scoliosis. In three children the curve pattern changed during follow-up. CONCLUSIONS: This report of idiopathic scoliosis in a father and six children supports the present theories on a genetic origin of the disorder but also stresses the importance of dynamic processes in the development of scoliosis. PMID- 11493868 TI - Bacteroides fragilis vertebral osteomyelitis secondary to anal dilatation. AB - STUDY DESIGN: A case report of anaerobic vertebral osteomyelitis after anal dilatation. OBJECTIVES: To present a patient with monomicrobial anaerobic vertebral osteomyelitis secondary to a previously undescribed source of infection. SUMMARY OF BACKGROUND DATA: A 17-year-old boy presented with low back pain 3 months after anal dilatation. METHODS: Physical examination, technetium 99m bone scan, plain radiograph, CT, and MRI studies of the lumbar spine were used to clinically diagnose lumbar osteomyelitis. Culture material from the involved disc was positive for Bacteroides fragilis. RESULTS: The patient recovered after 8 weeks of treatment with oral metronidazole. CONCLUSIONS: Bacteroides fragilis hematogenous osteomyelitis is a rare entity. This is the first reported case of such disease after anal dilatation. PMID- 11493869 TI - [Small acoustic neuroma: observation or treatment? A review of 207 cases]. AB - The management of small acoustic neuromas (AN) whether localized in an intracanalar position (stage I) or with a small extension of less then 2 cm into the cerebellopontine angle (stage II) remains under debate. Proposed strategies include surgery, stereotactic irradiation and observation. From 1987 to 1997, among 343 AN referred to our department, 207 were small (83 stage I and 124 stage II). Initially, 72 patients were treated conservatively mainly because of their age (over 60-65) and 132 were operated on. Three patients underwent irradiation because of their poor general condition. Significant tumor growth was observed in 15 cases in the conservative treatment group; 14 of these patients underwent secondary surgery and one irradiation. Among the 146 AN operated on (132 initially and 14 secondarily), 142 small AN were operated on via a transpetrosal approach (64% translabyrinthine, 21% middle-fossa and 15% retrosigmoid) and 4 AN, which became large tumors during the observation period, were treated through the translabyrinthine approach. No mortality was observed in our series. Postoperative complications included 11 CSF leakages necessitating reoperation (8%). In 93% of the cases, postoperative facial function at one year was good. Hearing preservation was attempted in 51 selected cases (pure tone average=50 dB, speech discrimination score 100%) with a 51% success rate (53% and 48% through middle-fossa and retrosigmoid approaches respectively). In our opinion, surgery with this risk-benefit ratio is indicated for small AN, except in the elderly for whom conservative management is preferred and in patients in poor clinical condition with a growing AN, for whom irradiation is recommended. PMID- 11493870 TI - [Diagnostic problems in brain abscess: 45 cases]. AB - BACKGROUND AND PURPOSE: We analysed the difficulties encountered in the differential diagnosis between brain abscess and brain tumor and their influence on treatment and outcome. METHODS: - Forty-five adults with brain abscess operated on between 1993 and 1999 were retrospectively reviewed. We studied preoperative diagnosis, clinical, radiological, bacteriological findings, surgical procedure, primary sources of infection and outcome. RESULTS: Preoperative diagnosis was right in 55.6% (25/45), wrong in 22.2% (10/45) and doubtful in 22.2% (10/45). Diffusion-weighted MR imaging was successfully used in 4 doubtful cases to make the differential diagnosis between abscess and tumor. When the preoperative diagnosis was right, the surgical procedure was a burr-hole aspiration in 73.3% (22/25) whereas when it was wrong, an excision was performed in 60% (6/10) of the cases. Aspiration was the last diagnostic investigation in 80% (8/10) of doubtful cases. Microbacterial organisms were identified in 75.5% (34/45) of the cases and primary cause of infection in 62.2% (28/45). The outcome depended on clinical status on admission, preoperative diagnosis and surgical procedure. In four cases, diffusion-weighted MRI allowed differential diagnosis between brain abscess and tumor through calculation of the Apparent Diffusion Coefficient which is low in abscess and high in cystic tumor. CONCLUSION: The diagnosis of brain abscess remains difficult in certain patients. Correct preoperative diagnosis influences the decision on the appropriate surgical procedure and helps improve outcome. PMID- 11493871 TI - [Factors influencing recurrence and role of radiotherapy in filum terminale ependymomas. 14 cases and review of the literature]. AB - PURPOSE: We reviewed a series of 14 cases of filum terminale ependymoma and 264 cases in the literature, to study the characteristics of these tumors and specifically to determine factors influencing recurrence. MATERIAL AND METHODS: This series analyzed data between 1984 and 1998. The mean follow-up period was 5.5 years and the median age 40 years. Pain was the first symptom, except in one case, with progressive sensitive-motor deficit. Four patients had a myelography and a CT scan, and ten a MRI. All patients were operated with a total resection in 12 cases. All tumors, except one, were a myxopapillary ependymoma. RESULTS: Clinical results were excellent in 2 cases, good in 8, stable in 2 and worse in 2. Two patients had a recurrence after an initial sub-total resection, both were operated on again, followed by post-operative radiotherapy. Among the 278 filum terminale ependymoma, removal was total in 200 (72%). A recurrence occurred in 15% of them after total removal, and in 43% after partial removal (p<0.001). Among patients with partial removal, recurrence was observed in 33% of them if they had post-operative radiotherapy, and in 55% of them if they did not have post-operative radiotherapy (p<0.05). CONCLUSION: The extent of tumor removal has a statistically significant effect upon recurrence. This review is in favor of post-operative radiotherapy in case of partial removal, but this systematic attitude can be discussed after a critical analysis of this study. Long term follow-up is mandatory due to the possibility of late recurrence. PMID- 11493872 TI - [Spondylodiscitis due to Histoplasma duboisii. Report of two cases and review of the literature]. AB - We report two cases of African histoplasmosis caused by Histoplasma duboisii located in the spine, on two Malian adults aged 40 and 47 years respectively. Clinical signs consisted of dorsal spinal syndrome in both cases and a paraplegia in one case. Radiography and CT scan revealed a spondylodiscitis without any bone reconstruction in both cases and a prevertebral abscess in one case. Diagnosis was confirmed by mycological and pathological examinations of surgical specimen which revealed Histoplasma duboisii fungus. Medical treatment, after surgical repair, consisted in ketoconazola with positive results after 12 months. This localisation is rare. It raises diagnosis problems with tuberculous spondylodiscitis because of their radio-clinical resemblance. PMID- 11493873 TI - [Diabetes insipidus after endoscopic third ventriculostomy. A case report and review of the literature]. AB - Endoscopic third ventriculostomy has been found to be successful for treating occlusive hydrocephalus. The complication rate ranges from 6 to 12%. Intraoperative bleeding is the most common incident. Endocrinological failures are rare, mainly due to the proximity of the hypothalamic structures. We report the case of a 33-year-old man who was referred in emergency for subacute hydrocephalus related to a tentorium meningioma. The hydrocephalus was treated by endoscopic third ventriculostomy. During the procedure, the floor of the third ventricle was found to be thick but fenestration was performed without incident. After surgery, the clinical signs of hydrocephalus disappeared but diabetes insipidus was diagnosed the same day. There were no other endocrinology disorders. Medical treatment with vasopressin allowed resolution of the diabetes insipidus in fifteen days. Surgical debulking of the meningioma was then achieved via a subtemporal approach. There was no recurrence of the endocrinology disorder. Diabetes insipidus is an unpredictable complication of third ventriculostomy. The mechanism is not well known. It is however a transient disorder that can easily be treated with vasopresin and therefore should not modify the indications of third ventriculostomy, especially in tumor-related hydrocephalus. PMID- 11493874 TI - [Intramedullary spinal cord sarcoidosis. Case report and review of the literature]. AB - Sarcoidosis is a multisystemic granulomatosis disease of unknown etiology which commonly involves hilary nodes and lungs. Involvement of the nervous system represents 5 to 15% of the cases. We report on a rare case of cervicothoracic spinal cord sarcoidosis in an African man and review the literature on the subject. His only symptoms were those of a subacute myelopathy. On MR-imaging, coexistence of abnormal signal of cervicothoracic spinal cord with mediastinal lymph nodes was suggestive of sarcoidosis, which was confirmed by bronchial biopsies making unnecessary biopsies of the spinal cord lesion. Under corticotherapy both symptoms and radiological abnormalities rapidly regressed. We emphasize the rarity of sarcoidosis exclusively revealed by a myelopathy. We stress the importance of searching other localizations which are easier and safer to biopsy than the spinal cord lesion itself and illustrate the MR appearances of the lesion which are suggestive, however not specific of the disease. PMID- 11493875 TI - [Spontaneous regression of paraplegia caused by spontaneous cervico-thoracic epidural hematoma]. AB - Spontaneous spinal epidural hematoma is a rare condition that usually requires surgical treatment in emergency. We report the unusual and well-documented case of a spontaneous spinal epidural hematoma, extending from C6 to T2, occurring in a 56-year old man and causing paraplegia. Without surgery, the paraplegia spontaneously and completely recovered within the first 24 hours. Serial MRI studies demonstrated the remitting course of the spinal epidural hematoma. Emergency surgical evacuation should still be the standard treatment for patients with disabling and/or persistent neurologic deficit. A conservative approach under close neurologic observation may be recommended for patients with no or mild deficits, for patients who show early and continuous clinical improvement within the first 24 hours and for patients with small and/or non-compressive spinal epidural hematoma. Reported cases of spontaneous remission are very rare. PMID- 11493876 TI - [Central neurocytoma revealed by intraventricular hemorrhage. A case report and review of the literature]. AB - A case of intraventricular tumor presenting with intraventricular hemorrhage in a 38-year-old man is reported. Initial symptoms were an acute onset of headache with projectile vomiting. CT-scan revealed intraventricular hemorrhage. Cerebral angiography did not show any vascular malformation. MRI study, performed initially and several days after, disclosed and confirmed the presence of an intraventricular mass. The patient was operated on via a transfrontal route, and a complete tumor removal was achieved. Pathological examination was consistent with a central neurocytoma. Two similar cases from the literature are discussed with respect to differential diagnosis and practical management considerations. PMID- 11493877 TI - A sound approach to GM debate. PMID- 11493879 TI - Drugs firms inflate research costs, watchdog says. PMID- 11493878 TI - Mad-cow outbreak spurs German drive to combat prion diseases. PMID- 11493880 TI - Medical journals seek means to free authors from industry. PMID- 11493881 TI - Golf course threatens to leave hole in fossil records. PMID- 11493882 TI - Canada plans to give unified voice to science. PMID- 11493883 TI - Commission plots transgenic future. PMID- 11493884 TI - Whistle-blowers wait for overbilling verdict. PMID- 11493886 TI - Senate urges Bush to act on climate change. PMID- 11493887 TI - Enterprising drug company offers cash for chemicals. PMID- 11493889 TI - Down and out in Murray Hill. PMID- 11493890 TI - Faster, better, cheaper genotyping. PMID- 11493891 TI - Finding the right questions to ask about the lives of human clones. PMID- 11493892 TI - Why are Indian journals' impact factors so low? PMID- 11493898 TI - Avoiding ambiguity. PMID- 11493893 TI - Genome helpdesk site keeps information public. PMID- 11493899 TI - Design by numbers. PMID- 11493900 TI - Nifty nanoplankton. PMID- 11493901 TI - Molecular electronics. Momentous period for nanotubes. PMID- 11493903 TI - Genome sequencing. The ABC of symbiosis. PMID- 11493904 TI - Chemistry. On the threshold of stability. PMID- 11493905 TI - Signal transduction. Barriers come down. PMID- 11493907 TI - Pattern of focal gamma-bursts in chess players. PMID- 11493908 TI - Anabolism. Low mechanical signals strengthen long bones. AB - Although the skeleton's adaptability to load-bearing has been recognized for over a century, the specific mechanical components responsible for strengthening it have not been identified. Here we show that after mechanically stimulating the hindlimbs of adult sheep on a daily basis for a year with 20-minute bursts of very-low-magnitude, high-frequency vibration, the density of the spongy (trabecular) bone in the proximal femur is significantly increased (by 34.2%) compared to controls. As the strain levels generated by this treatment are three orders of magnitude below those that damage bone tissue, this anabolic, non invasive stimulus may have potential for treating skeletal conditions such as osteoporosis. PMID- 11493909 TI - Vision. Realignment of cones after cataract removal. AB - Through unique observations of an adult case of bilateral congenital cataract removal, we have found evidence that retinal photoreceptors will swiftly realign towards the brightest regions in the pupils of the eye. Cones may be phototropic, actively orientating themselves towards light like sunflowers in a field. PMID- 11493910 TI - Palaeoceanography. Antarctic stratification and glacial CO2. AB - One way of accounting for lowered atmospheric carbon dioxide concentrations during Pleistocene glacial periods is by invoking the Antarctic stratification hypothesis, which links the reduction in CO2 to greater stratification of ocean surface waters around Antarctica. As discussed by Sigman and Boyle, this hypothesis assumes that increased stratification in the Antarctic zone (Fig. 1) was associated with reduced upwelling of deep waters around Antarctica, thereby allowing CO2 outgassing to be suppressed by biological production while also allowing biological production to decline, which is consistent with Antarctic sediment records. We point out here, however, that the response of ocean eddies to increased Antarctic stratification can be expected to increase, rather than reduce, the upwelling rate of deep waters around Antarctica. The stratification hypothesis may have difficulty in accommodating eddy feedbacks on upwelling within the constraints imposed by reconstructions of winds and Antarctic-zone productivity in glacial periods. PMID- 11493912 TI - Structure of the Ku heterodimer bound to DNA and its implications for double strand break repair. AB - The Ku heterodimer (Ku70 and Ku80 subunits) contributes to genomic integrity through its ability to bind DNA double-strand breaks and facilitate repair by the non-homologous end-joining pathway. The crystal structure of the human Ku heterodimer was determined both alone and bound to a 55-nucleotide DNA element at 2.7 and 2.5 A resolution, respectively. Ku70 and Ku80 share a common topology and form a dyad-symmetrical molecule with a preformed ring that encircles duplex DNA. The binding site can cradle two full turns of DNA while encircling only the central 3-4 base pairs (bp). Ku makes no contacts with DNA bases and few with the sugar-phosphate backbone, but it fits sterically to major and minor groove contours so as to position the DNA helix in a defined path through the protein ring. These features seem well designed to structurally support broken DNA ends and to bring the DNA helix into phase across the junction during end processing and ligation. PMID- 11493913 TI - History of trace gases in presolar diamonds inferred from ion-implantation experiments. AB - Diamond grains are the most abundant presolar grains found in primitive meteorites. They formed before the Solar System, and therefore provide a record of nuclear and chemical processes in stars and in the interstellar medium. Their origins are inferred from the unusual isotopic compositions of trace elements mainly xenon-which suggest that they came from supernovae. But the exact nature of the sources has been enigmatic, as has the method by which noble gases were incorporated into the grains. One observation is that different isotopic components are released at different temperatures when the grains are heated, and it has been suggested that these components have different origins. Here we report results of a laboratory study that shows that ion implantation (previously suggested on other grounds) is a viable mechanism for trapping noble gases. Moreover, we find that ion implantation of a single isotopic composition can produce both low- and high-temperature release peaks from the same grains. We conclude that both isotopically normal and anomalous gases may have been implanted by multiple events separated in space and/or time, with thermal processing producing an apparent enrichment of the anomalous component in the high-temperature release peak. The previous assumption that the low- and high temperature components were not correlated may therefore have led to an overestimate of the abundance of anomalous argon and krypton, while obscuring an enhancement of the light-in addition to the heavy-krypton isotopes. PMID- 11493914 TI - Two-dimensional imaging of electronic wavefunctions in carbon nanotubes. AB - The drive towards the development of molecular electronics is placing increasing demands on the level of control that must be exerted on the electronic structure of materials. Proposed device architectures ultimately rely on tuning the interactions between individual electronic states, which amounts to controlling the detailed spatial structure of the electronic wavefunctions in the constituent molecules. Few experimental tools are available to probe this spatial structure directly, and the shapes of molecular wavefunctions are usually only known from theoretical investigations. Here we present scanning tunnelling spectroscopy measurements of the two-dimensional structure of individual wavefunctions in metallic single-walled carbon nanotubes; these measurements reveal spatial patterns that can be directly understood from the electronic structure of a single graphite sheet, and which represent an elegant illustration of Bloch's theorem at the level of individual wavefunctions. We also observe energy dependent interference patterns in the wavefunctions and exploit these to directly measure the linear electronic dispersion relation of the metallic single walled carbon nanotube. PMID- 11493915 TI - Observation of individual vortices trapped along columnar defects in high temperature superconductors. AB - Many superconductors do not entirely expel magnetic flux-rather, magnetic flux can penetrate the superconducting state in the form of vortices. Moving vortices create resistance, so they must be 'pinned' to permit dissipationless current flow. This is a particularly important issue for the high-transition-temperature superconductors, in which the vortices move very easily. Irradiation of superconducting samples by heavy ions produces columnar defects, which are considered to be the optimal pinning traps when the orientation of the column coincides with that of the vortex line. Although columnar defect pinning has been investigated using macroscopic techniques, it has hitherto been impossible to resolve individual vortices intersecting with individual defects. Here we achieve the resolution required to image vortex lines and columnar defects in Bi2Sr2CaCu2O8+delta (Bi-2212) thin films, using a 1-MV field-emission electron microscope. For our thin films, we find that the vortex lines at higher temperatures are trapped and oriented along tilted columnar defects, irrespective of the orientation of the applied magnetic field. At lower temperatures, however, vortex penetration always takes place perpendicular to the film plane, suggesting that intrinsic 'background' pinning in the material now dominates. PMID- 11493916 TI - Vacancies in solids and the stability of surface morphology. AB - Determining how thermal vacancies are created and destroyed in solids is crucial for understanding many of their physical properties, such as solid-state diffusion. Surfaces are known to be good sources and sinks for bulk vacancies, but directly determining where the exchange between the surface and the bulk occurs is difficult. Here we show that vacancy generation (and annihilation) on the (110) surface of an ordered nickel-aluminium intermetallic alloy does not occur over the entire surface, but only near atomic step edges. This has been determined by oscillating the sample's temperature and observing in real time the response of the surface structure as a function of frequency (a version of Angstrom's method of measuring thermal conductivity) using low-energy electron microscopy. Although the surface-exchange process is slow compared with bulk diffusion, the vacancy-generation rate nevertheless controls the dynamics of the alloy surface morphology. These observations, demonstrating that surface smoothing can occur through bulk vacancy transport rather than surface diffusion, should have important implications for the stability of fabricated nanoscale structures. PMID- 11493917 TI - Generation and characterization of a fairly stable triplet carbene. AB - Most molecules are held together by covalent bonds-electron pairs jointly shared by the two atoms that are linked by the bond. Free radicals, in contrast, have at least one unpaired electron. In the case of carbon-based radicals, the carbon atom at the radical centre no longer makes four bonds with other atoms as it would do in its normal, tetravalent state. The presence of unpaired electrons renders such radicals highly reactive, so they normally occur only as transient intermediates during chemical reactions. But the discovery by Gomberg in 1900 of triphenylmethyl, the first relatively stable free radical containing a central trivalent carbon atom, illustrated that radicals with suitable geometrical and electronic structures can be stable. Compounds containing a divalent carbon atom that uses only two of its four valence electrons for bonding are usually less stable than Gomberg-type radicals with trivalent carbon. Although the role of these so-called carbenes in chemical reactions has long been postulated, they were unambiguously identified only in the 1950s. More recently, stable carbenes have been prepared, but the singlet state of these molecules, with the two nonbonding valence electrons paired, means that they are not radicals. Carbenes in the second possible electronic state, the triplet state, are radicals: the two nonbonding electrons have parallel spins and occupy different orbitals. Here we report the preparation and characterization of a triplet carbene, whose half-life of 19 minutes at room temperature shows it to be significantly more stable than previously observed triplet carbenes. PMID- 11493918 TI - Normal faulting in central Tibet since at least 13.5 Myr ago. AB - Tectonic models for the evolution of the Tibetan plateau interpret observed east west thinning of the upper crust to be the result of either increased potential energy of elevated crust or geodynamic processes that may be unrelated to plateau formation. A key piece of information needed to evaluate these models is the timing of deformation within the plateau. The onset of normal faulting has been estimated to have commenced in southern Tibet between about 14 Myr ago and about 8 Myr ago and, in central Tibet, about 4 Myr ago. Here, however, we report a minimum age of approximately 13.5 Myr for the onset of graben formation in central Tibet, based on mineralization ages determined with Rb-Sr and 40Ar-39Ar data that post-date a major graben-bounding normal fault. These data, along with evidence for prolonged activity of normal faulting in this and other Tibetan grabens, support models that relate normal faulting to processes occurring beneath the plateau. Thinning of the upper crust is most plausibly the result of potential-energy increases resulting from spatially and temporally heterogeneous changes in thermal structure and density distribution within the crust and upper mantle beneath Tibet. This is supported by recent geophysical and geological data, which indicate that spatial heterogeneity exists in both the Tibetan crust and lithospheric mantle. PMID- 11493919 TI - Resistance to mantle flow inferred from the electromagnetic strike of the Australian upper mantle. AB - Seismic anisotropy is thought to result from the strain-induced lattice-preferred orientation of mantle minerals, especially olivine, owing to shear waves propagating faster along the a-axis of olivine crystals than along the other axes. This anisotropy results in birefringence, or 'shear-wave splitting', which has been investigated in numerous studies. Although olivine is also anisotropic with respect to electrical conductivity (with the a-axis being most conductive), few studies of the electrical anisotropy of the upper mantle have been undertaken, and these have been limited to relatively shallow depths in the lithospheric upper mantle. Theoretical models of mantle flow have been used to infer that, for progressive simple shear imparted by the motion of an overriding tectonic plate, the a-axes of olivine crystals should align themselves parallel to the direction of plate motion. Here, however, we show that a significant discrepancy exists between the electromagnetic strike of the mantle below Australia and the direction of present-day absolute plate motion. We infer from this discrepancy that the a-axes of olivine crystals are not aligned with the direction of the present-day plate motion of Australia, indicating resistance to deformation of the mantle by plate motion. PMID- 11493920 TI - Unicellular cyanobacteria fix N2 in the subtropical North Pacific Ocean. AB - Fixed nitrogen (N) often limits the growth of organisms in terrestrial and aquatic biomes, and N availability has been important in controlling the CO2 balance of modern and ancient oceans. The fixation of atmospheric dinitrogen gas (N2) to ammonia is catalysed by nitrogenase and provides a fixed N for N-limited environments. The filamentous cyanobacterium Trichodesmium has been assumed to be the predominant oceanic N2-fixing microorganism since the discovery of N2 fixation in Trichodesmium in 1961 (ref. 6). Attention has recently focused on oceanic N2 fixation because nitrogen availability is generally limiting in many oceans, and attempts to constrain the global atmosphere-ocean fluxes of CO2 are based on basin-scale N balances. Biogeochemical studies and models have suggested that total N2-fixation rates may be substantially greater than previously believed but cannot be reconciled with observed Trichodesmium abundances. It is curious that there are so few known N2-fixing microorganisms in oligotrophic oceans when it is clearly ecologically advantageous. Here we show that there are unicellular cyanobacteria in the open ocean that are expressing nitrogenase, and are abundant enough to potentially have a significant role in N dynamics. PMID- 11493921 TI - Density-dependent mortality in an oceanic copepod population. AB - Planktonic copepods are primary consumers in the ocean and are perhaps the most numerous metazoans on earth. Secondary production by these zooplankton supports most food webs of the open sea, directly affecting pelagic fish populations and the biological pump of carbon into the deep ocean. Models of marine ecosystems are quite sensitive to the formulation of the term for zooplankton mortality, although there are few data available to constrain mortality rates in such models. Here we present the first evidence for nonlinear, density-dependent mortality rates of open-ocean zooplankton. A high-frequency time series reveals that per capita mortality rates of eggs of Calanus finmarchicus Gunnerus are a function of the abundance of adult females and juveniles. The temporal dynamics of zooplankton populations can be influenced as much by time-dependent mortality rates as by variations in 'bottom up' forcing. The functional form and rates chosen for zooplankton mortality in ecosystem models can alter the balance of pelagic ecosystems, modify elemental fluxes into the ocean's interior, and modulate interannual variability in pelagic ecosystems. PMID- 11493922 TI - Erythropoietin-mediated neuroprotection involves cross-talk between Jak2 and NF kappaB signalling cascades. AB - Erythropoietin, a kidney cytokine regulating haematopoiesis (the production of blood cells), is also produced in the brain after oxidative or nitrosative stress. The transcription factor hypoxia-inducible factor-1 (HIF-1) upregulates EPO following hypoxic stimuli. Here we show that preconditioning with EPO protects neurons in models of ischaemic and degenerative damage due to excitotoxins and consequent generation of free radicals, including nitric oxide (NO). Activation of neuronal EPO receptors (EPORs) prevents apoptosis induced by NMDA (N-methyl-d-aspartate) or NO by triggering cross-talk between the signalling pathways of Janus kinase-2 (Jak2) and nuclear factor-kappaB (NF-kappaB). We show that EPOR-mediated activation of Jak2 leads to phosphorylation of the inhibitor of NF-kappaB (IkappaB), subsequent nuclear translocation of the transcription factor NF-kappaB, and NF-kappaB-dependent transcription of neuroprotective genes. Transfection of cerebrocortical neurons with a dominant interfering form of Jak2 or an IkappaBalpha super-repressor blocks EPO-mediated prevention of neuronal apoptosis. Thus neuronal EPORs activate a neuroprotective pathway that is distinct from previously well characterized Jak and NF-kappaB functions. Moreover, this EPO effect may underlie neuroprotection mediated by hypoxic ischaemic preconditioning. PMID- 11493923 TI - Spred is a Sprouty-related suppressor of Ras signalling. AB - Cellular proliferation, and differentiation of cells in response to extracellular signals, are controlled by the signal transduction pathway of Ras, Raf and MAP (mitogen-activated protein) kinase. The mechanisms that regulate this pathway are not well known. Here we describe two structurally similar tyrosine kinase substrates, Spred-1 and Spred-2. These two proteins contain a cysteine-rich domain related to Sprouty (the SPR domain) at the carboxy terminus. In Drosophila, Sprouty inhibits the signalling by receptors of fibroblast growth factor (FGF) and epidermal growth factor (EGF) by suppressing the MAP kinase pathway. Like Sprouty, Spred inhibited growth-factor-mediated activation of MAP kinase. The Ras-MAP kinase pathway is essential in the differentiation of neuronal cells and myocytes. Expression of a dominant negative form of Spred and Spred-antibody microinjection revealed that endogenous Spred regulates differentiation in these types of cells. Spred constitutively associated with Ras but did not prevent activation of Ras or membrane translocation of Raf. Instead, Spred inhibited the activation of MAP kinase by suppressing phosphorylation and activation of Raf. Spred may represent a class of proteins that modulate Ras-Raf interaction and MAP kinase signalling. PMID- 11493924 TI - General transcription factors bind promoters repressed by Polycomb group proteins. AB - To maintain cell identity during development and differentiation, mechanisms of cellular memory have evolved that preserve transcription patterns in an epigenetic manner. The proteins of the Polycomb group (PcG) are part of such a mechanism, maintaining gene silencing. They act as repressive multiprotein complexes that may render target genes inaccessible to the transcriptional machinery, inhibit chromatin remodelling, influence chromosome domain topology and recruit histone deacetylases (HDACs). PcG proteins have also been found to bind to core promoter regions, but the mechanism by which they regulate transcription remains unknown. To address this, we used formaldehyde-crosslinked chromatin immunoprecipitation (X-ChIP) to map TATA-binding protein (TBP), transcription initiation factor IIB (TFIIB) and IIF (TFIIF), and dHDAC1 (RPD3) across several Drosophila promoter regions. Here we show that binding of PcG proteins to repressed promoters does not exclude general transcription factors (GTFs) and that depletion of PcG proteins by double-stranded RNA interference leads to de-repression of developmentally regulated genes. We further show that PcG proteins interact in vitro with GTFs. We suggest that PcG complexes maintain silencing by inhibiting GTF-mediated activation of transcription. PMID- 11493930 TI - Q & A about this issue. PMID- 11493925 TI - A Drosophila Polycomb group complex includes Zeste and dTAFII proteins. AB - A goal of modern biology is to identify the physical interactions that define 'functional modules' of proteins that govern biological processes. One essential regulatory process is the maintenance of master regulatory genes, such as homeotic genes, in an appropriate 'on' or 'off' state for the lifetime of an organism. The Polycomb group (PcG) of genes maintain a repressed transcriptional state, and PcG proteins form large multiprotein complexes, but these complexes have not been described owing to inherent difficulties in purification. We previously fractionated a major PcG complex, PRC1, to 20-50% homogeneity from Drosophila embryos. Here, we identify 30 proteins in these preparations, then further fractionate the preparation and use western analyses to validate unanticipated connections. We show that the known PcG proteins Polycomb, Posterior sex combs, Polyhomeotic and dRING1 exist in robust association with the sequence-specific DNA-binding factor Zeste and with numerous TBP (TATA-binding protein)-associated factors that are components of general transcription factor TFIID (dTAFIIs). Thus, in fly embryos, there is a direct physical connection between proteins that bind to specific regulatory sequences, PcG proteins, and proteins of the general transcription machinery. PMID- 11493931 TI - "Effectiveness" in the evaluation of new drugs: a misunderstood concept? PMID- 11493932 TI - Interactions between doctors and pharmaceutical sales representatives. PMID- 11493933 TI - Digestive Disease Week 2000 conference report: New treatments for chronic hepatitis C. AB - At the Digestive Disease Week (DDW) conference and 101st Meeting of the American Gastroenterological Association in San Diego, California, May 22 to 24, 2000, over 400 abstracts on hepatitis C were submitted for posters or oral presentations to the American Association for the Study of Liver Diseases. A substantial portion of the program discussed the treatment of chronic hepatitis C, focusing on interferon and ribavirin combination therapy, substitution of amantadine for ribavirin, and the use of pegylated interferons. In randomized, clinical trials, combination therapy with interferon- alpha-2B and ribavirin results in a greater sustained virilogical response than treatment with interferon alone. Combination therapy is generally safe and well tolerated, but there is a need to monitor patients throughout treatment for hemolytic side effects, depression, and weight and lipid profiles. In the present review some background information on chronic hepatitis C is given and some of the more relevant abstracts presented on this subject at the DDW conference are highlighted. PMID- 11493934 TI - Development of a standardized, comprehensive "ideal drug detail". AB - OBJECTIVES: To develop a standardized, comprehensive ideal drug detail for use in face-to-face education about individual drugs. METHODS: A random sample of 603 physicians and pharmacists was selected and stratified to include input from each of the following specialties: family practice, internal medicine, surgery, pediatrics, psychiatry, obstetrics/gynecology, geriatric medicine and clinical pharmacology. Thirty-one potential items were generated by the investigators from a preliminary survey of a local convenience sample of physicians and pharmacists. A modified Delphi consensus process was used in the large sample to determine which items should be included in the ideal drug detail. In each round of the Delphi process, respondents rated each item on a seven-point scale of importance and were then given feedback of the cumulative ratings for each item. Rounds were continued until consensus was obtained on all items. RESULTS: The response rate to the first round was 55.3%; 85.5% of these respondents responded to the second round. Response rates varied between specialties from 44% to 70%. Attempts to contact nonresponders to measure potential nonrespondent bias were unsuccessful. Consensus was obtained on 19 items after the first round, and on the remaining 12 items after the second round. Four items were dropped because they were unimportant. There was variation in modal response between specialties on eight items. CONCLUSIONS: Consensus was obtained among a sizable and interested sample of Canadian physicians and pharmacists on the items of information needed to prescribe a drug appropriately. Subsequent work will refine this list into a usable template to develop ideal drug details for specific drugs, to develop an assessment process to measure quality of information, and to assess the impact of this program on prescribing and patient outcomes. PMID- 11493935 TI - Improving prescribing in the elderly: a study in the long term care setting. AB - OBJECTIVES: To determine the prevalence and predictors of potentially inappropriate prescribing of medications in the long term care setting, and to determine the effectiveness of follow-up pharmacist letters to the prescribing physicians in improving prescribing. PATIENTS AND METHODS: The Improving Prescribing in the Elderly Tool was applied to the charts of all long term care patients aged 65 years and over at Parkwood Hospital, a rehabilitation hospital/long term care facility in London, Ontario. All potentially inappropriate prescriptions were verified by a consensus panel consisting of a family physician, a geriatric medicine specialist and a geriatric pharmacist. Follow-up letters to the prescribing physicians were developed that briefly described the concerns with the potentially inappropriate prescriptions and suggested safer alternatives. These letters were sent to the prescribing physicians, accompanied by a brief survey. Patient charts in which a potentially inappropriate prescription had been noted were reviewed for prescription changes two months after the prescribing physicians had received the follow-up letters. RESULTS: A total of 69 potentially inappropriate prescriptions were found in 65 of 355 long term care patients (18.3%). The most common types of potentially inappropriate prescriptions were anticholinergic drugs to manage antipsychotic side effects (17 cases), tricyclic antidepressants with active metabolites (16 cases), and long-acting benzodiazepines (14 cases). The total number of prescription medications (P<0.001), a history of mental illness (P=0.002) and a high minimum data set (MDS) score for depression (P=0.002) were all highly associated with potentially inappropriate prescribing. Variables that were not correlated with increased rates of potentially inappropriate prescribing included age, sex, code status, a diagnosis of dementia (as documented explicitly in the chart), high MDS scores for delirium or cognitive impairment, the date of the prescribing physician's graduation and the total Charlson comorbidity index score. Potentially inappropriate prescriptions were significantly less common in patients seen by a geriatric medicine specialist (P<0.001). In response to the follow-up letter suggesting safer alternatives, 37.9% of potentially inappropriate prescriptions were changed by the prescribing physician. Ninety-two per cent of responding physicians rated the follow-up letter as a "somewhat" or "very" helpful method for improving prescribing in elderly patients. CONCLUSIONS: Potentially inappropriate prescribing in the long term care setting is common and can be improved by the provision of a follow-up letter suggesting safer alternatives. PMID- 11493936 TI - Reporting of fatal adverse drug reactions. AB - OBJECTIVES: To review the reports of fatal adverse drug reactions (ADRs) submitted to the Ontario Medical Association Adverse Drug Reactions Monitoring Program between 1990 and 1994; to identify drugs associated with fatal outcomes; and to assess the causative role of the drug in these events and the completeness of the data in these reports. METHODS: Drug(s) identified on each ADR report as being responsible for the reaction were considered. Agents were classified by the Anatomical Therapeutic and Chemical classification system. The causality of each ADR report was evaluated by using an algorithmic rating scale. RESULTS: From the Ontario Medical Association database, 97 cases of ADRs that resulted in death were reviewed. One hundred fourteen medications were implicated as "suspect" drugs in the 97 deaths. The most commonly implicated drug classes were musculoskeletal agents, blood and blood-forming organ agents, and nervous system agents. Patients over 65 years of age comprised 60% of this series. After independent assessment as to causality, 13% of the cases were rated as probable, 86% were rated as possible and 1% were rated as doubtful. Seventy per cent of reports did not include information regarding medical history. Forty-two per cent of cases failed to provide adequate information to evaluate the feasibility of the time to onset of the ADR. The use of concomitant drugs was not reported in 12% of cases. CONCLUSIONS: The drugs most frequently implicated in fatal ADRs were consistent with those reported in other studies. Algorithmic causality assessments were of limited value in these reports. The completeness of the reports and adequacy of the information were poor. The type of reporting forms and information provided were not homogenous. There is a need to improve quality of reporting and harmonize reporting forms between monitoring bodies. The feasibility of unique data collection forms and obligatory reporting for fatal ADRs should be considered. PMID- 11493937 TI - Motherisk alert: Folic acid fortification of flour--three years later. AB - Fortification of flour with folic acid aims to prevent neural tube defects. In Canada, flour fortification began in 1997. The objective of this study was to quantify the changes in women's erythrocyte folic acid levels. Comparing these levels among normocytic women revealed an almost twofold increase (from 517+/-215 nmol/L in 1995 to 901+/-318 nmol/L in 1998) (P<0.00001). While the mean value increased substantially, even now, women at the low range of measured values (367 nmol/L) confer a relative risk of 3.2 for neural tube defects. For these women, higher intake of nutritional folic acid and perinatal supplementation of folic acid tablets continue to be crucial. PMID- 11493938 TI - Is chronic poisoning with acetaminophen in children a frequent occurrence in Toronto? AB - BACKGROUND: Acetaminophen is a common cause of poisoning in children. Recent American studies suggest that acetaminophen poisonings pose serious risks in children, particularly in the case of chronic poisoning caused by therapeutic error. OBJECTIVE: To evaluate whether chronic acetaminophen poisoning in children is a frequent occurrence in a large, Canadian, urban population. PATIENTS AND METHODS: Retrospective study. Charts of all patients admitted to The Hospital for Sick Children, Toronto, Ontario from January 1, 1990 to June 31, 1996 with an acetaminophen overdose were reviewed. RESULTS: A total of 110 patients were admitted within the study period; only four of whom were preschool children (younger than five years of age). Among the preschool children, three had an acute overdose and one had possible chronic poisoning by therapeutic error. All preschool children were treated with N-acetylcysteine; one developed hepatotoxicity (aspartate aminotransferase or alanine aminotransferase greater than 1000 U/L) after presenting 24 h after acute ingestion. Of the remaining patients, all were adolescents; 102 had acute intentional overdose and four had staggered intentional overdoses. Fifty-three adolescents were treated with N acetyl cysteine. Hepatotoxicity was present in 13 of 63 adolescents (21%). No patients required liver transplantation or died. CONCLUSIONS: Contrary to American experience, chronic acetaminophen poisoning, including therapeutic error in children in Toronto, is a rare occurrence--most cases of acetaminophen poisonings are acute intentional ingestion in adolescents. PMID- 11493939 TI - Metformin-associated lactic acidosis in a low risk patient. AB - Metformin is an oral hypoglycemic agent belonging to the class of biguanides that are commonly used in the treatment of type II diabetes mellitus. Lactic acidosis is a rare but severe adverse reaction that occurs primarily in patients with contraindications such as renal failure. The case of a 71-year-old woman with type II diabetes, in whom severe metformin-associated lactic acidosis was precipitated by acute renal failure in the absence of pre-existing chronic renal failure or other absolute contraindications to biguanide use, is presented. Aggressive correction of the acidosis and prolonged dialysis resulted in a favourable outcome despite severe acidosis. The present case report shows that metformin-associated lactic acidosis can occur in patients without pre-existing renal insufficiency. Metformin should be temporarily stopped when acute renal failure occurs or is anticipated. PMID- 11493940 TI - Steroid-induced mania in an adolescent: risk factors and management. AB - Few studies have clearly defined the risk factors and management of steroid induced mania, particularly in children and adolescents. The case study of a 15 year-old woman with steroid-induced mania is examined to delineate risk factors and management. Genetic predisposition to mood disorders, previous exposure to steroids and the concomitant use of clarithromycin are potential risk factors. Mood stabilizers such as lithium and/or valproic acid may be useful in the acute and prophylactic treatment of steroid-induced mania. PMID- 11493942 TI - H pylori eradication and GERD. PMID- 11493943 TI - Methotrexate and maintenance of remission in Crohn's disease. PMID- 11493944 TI - Droperidol: an effective adjuvant for difficult cases of conscious sedation? PMID- 11493945 TI - The sigmoidoscope: used too much or too little? PMID- 11493946 TI - Diabetes mellitus after liver transplantation: another extrahepatic manifestation of hepatitis C. PMID- 11493947 TI - Efficacy of lamivudine in decompensated cirrhosis secondary to hepatitis B: cautious optimism in the war not yet won. PMID- 11493948 TI - Hemodynamic characterization of arterialized and nonarterialized liver transplants in the rat. AB - Persistent hyperkinetic circulation after liver transplantation has been described in humans, but similar changes have not been well characterized in the rat model. This study aimed to investigate the hemodynamics of the systemic and splanchnic circulations in both arterialized and nonarterialized hepatic allografts. Orthotopic liver transplantation was performed in four groups of Sprague-Dawley rats. Group A comprised sham-operated rats with hepatic artery ligation that did not receive transplants; group B comprised rats that received transplants without arterialization; group C comprised sham-operated rats with intact hepatic artery that did not receive transplants; and group D comprised rats that received transplants with arterialization. Blood flow measurements were performed three weeks after the surgical procedure, using the radioactive microsphere method. The results showed that rats that received transplants exhibited a significantly higher cardiac index and lower systemic vascular resistance than the control rats. Splanchnic hyperemia was also present with increased mesenteric blood flow. However, there was no difference in hemodynamics between rats that received arterialized transplants and those that received nonarterialized transplants. Arterial collateral vessels from adjacent tissues were observed in the nonarterialized grafts; this was confirmed histologically. It is concluded that rats that undergo orthotopic liver transplantation exhibit hyperdynamic circulation, regardless of the arterial reconstruction procedure, possibly due to extensive collateral formation in the hepatosplanchnic circulation. PMID- 11493949 TI - Colorectal cancer screening: video-reviewed flexible sigmoidoscopy by nurse endoscopists--a Canadian community-based perspective. AB - BACKGROUND: Colorectal cancer (CRC) is the third most common incident cancer and the second most fatal cancer in Canada. Flexible sigmoidoscopy (FS) is one of the modalities under consideration for CRC screening. The present series reports on a screening program of FS performed by nonphysician endoscopists in a Canadian community setting, with video review of procedures by physicians and recommendation of follow-up colonoscopy where polyps are identified. RESULTS: Five hundred twenty-five, average-risk, asymptomatic patients were examined. After exclusion of inappropriate referrals, 488 remained for analysis. The duration and extent of examination were comparable with those of previous studies elsewhere. Compliance with suggested follow-up was 97.3%. Polyps were identified at FS in 15.4% of examinees. In 8.2% of patients, the polyps were neoplastic at subsequent histology. Four malignant lesions were detected, all at an early stage. There were no complications of FS. INTERPRETATION: This report shows that FS can be carried out safely and effectively by nonphysician personnel in a community setting in Canada. The manpower cost for nonphysician operators is considerably less than that for specialist physician endoscopists. This approach deserves consideration in cost effectiveness analyses of CRC screening. PMID- 11493950 TI - Small bowel review: Part II. AB - In the past year, there have been many advances in the area of small bowel physiology and pathology. In preparation for this review, over 1500 papers were assessed. Some have been selected and reviewed, with a particular focus on presenting clinically useful information for the practising gastroenterologist. Relevant review articles have been highlighted, and important clinical learning points have been stressed. The topics are varied in scope and wherever possible show a logical progression from basic physiology to pathophysiology to clinical disorders and management. PMID- 11493951 TI - Gastric cancer: past, present and future. AB - Gastric cancer remains a major cause of cancer mortality in the world. However, in the past 10 decades, the view of gastric cancer has been changing. This includes the unexplained decline in the incidence of the cancer, the proximal shift of the cancer in the stomach, the identification of Helicobacter pylori as an etiological agent, rapid development in molecular tumour biology, new treatment modalities and the adoption of mass screening for prevention. This article reviews the changing views of gastric cancer and the latest developments. PMID- 11493952 TI - Topographic mapping of collagenous gastritis. AB - A 74-year-old woman was investigated for abdominal pain and diarrhea. Endoscopic examinations including biopsies of the stomach and colon demonstrated the typical subepithelial deposits characteristic of collagenous gastritis and collagenous colitis. Histochemical and ultrastructural methods confirmed the presence of collagen in the subepithelial deposits. The topographic distribution of these collagen deposits and their relationship to the inflammatory process in the stomach were then defined by endoscopic mapping and multiple site biopsies of the mucosa in the gastric body and antrum. These studies indicate that collagenous gastritis not only is distinctive, but also is a far more extensive and diffuse inflammatory process than has previously been appreciated. PMID- 11493953 TI - Solitary rectal ulcer syndrome presenting with rectal prolapse, severe mucorrhea and eroded polypoid hyperplasia: case report and review of the literature. AB - A case of solitary rectal ulcer syndrome in a 36-year-old woman presenting with severe, persistent mucorrhea and eroded polypoid hyperplasia as the predominant clinical features, who was ultimately noted to have symptoms of rectal prolapse, is presented. Endoscopically, she had multiple (50 to 60) small, whitish polypoid lesions in the rectum that were initially misinterpreted as being a carpeted villous adenoma, juvenile polyposis or atypical proctitis. The lesions were treated with argon plasma coagulation with resolution, but a solitary rectal ulcer developed. The patient then admitted to a history of massive rectal prolapse over the preceding six months and underwent surgical treatment. Severe mucorrhea as the presenting feature and the presence of multiple polypoid lesions consistent with a histological diagnosis of eroded polypoid hyperplasia make the present case unique. PMID- 11493955 TI - A Message from the New Editorial Team. PMID- 11493954 TI - Dysplasia-associated polypoid mucosal lesion in a pelvic pouch after restorative proctocolectomy for ulcerative colitis. AB - A 32-year-old man with ulcerative colitis had a colectomy for toxic colitis. Later, a rectal mucosectomy was performed along with the creation of a pelvic pouch. In 1998, approximately 10 years after this staged restorative proctocolectomy was completed, endoscopic examination of the pelvic pouch detected a small mucosal polypoid mass lesion. Although the lesion had the macroscopic appearance of an inflammatory polyp, microscopic sections of the resected lesion revealed dysplastic changes. Endoscopic polypectomy was performed to remove the lesion, and further histological surveillance examinations of the pelvic pouch have not detected additional dysplastic mucosal changes. PMID- 11493956 TI - The National Wilms Tumor Study Pathology Center Relocates to Hopkins. PMID- 11493957 TI - A Grand Transcontinental Event in Madeira. PMID- 11493958 TI - A Free Immunoquery Database is Made Available as a Website. PMID- 11493959 TI - Molecular Insights into the Histogenesis and Pathogenesis of Gastrointestinal Stromal Tumors. AB - Gastrointestinal stromal tumors (GISTs) have long been problematic in terms of classification and determination of prognosis. Recent studies have suggested that GISTs differentiate toward a phenotype resembling the interstitial cells of Cajal. This has led to the important discovery that activating mutations in the KIT receptor tyrosine kinase play an important role in the pathogenesis of GISTs. These findings have helped clarify the distinction between GISTs and other mesenchymal neoplasms of the gastrointestinal tract and may translate into an improved ability to predict biologic behavior, as well as suggesting possible avenues for rational drug design for the treatment of GISTs. Int J Surg Pathol 8(1):5-10, 2000 PMID- 11493960 TI - Dysplasia in Chronic Ulcerative Colitis: A Molecular Approach to Its Differential Diagnosis. AB - Ulcerative colitis-related epithelial dysplasia represents a premalignant lesion, which may lead to the development of ulcerative colitis-related adenocarcinoma. The proper management of this condition requires proctocolectomy before the acquisition of the invasive phenotype. Patients with chronic ulcerative colitis, however, like the general population, may develop sporadic colorectal adenomas not related to the presence of the inflammatory bowel disease. These adenomas are also characterized by epithelial dysplasia, but their detection carries very different clinical implications. It is therefore essential to distinguish between chronic ulcerative colitis-related dysplasia and sporadic colorectal adenomas. The gross and histologic features do not provide a definite distinction between these two different types of dysplasia of the colonic epithelium. Recent developments in the molecular genetics may provide the necessary means. Int J Surg Pathol 8(1):11-16, 2000 PMID- 11493961 TI - Fine Needle Aspiration Biopsies of the Head and Neck: The Surgical Pathologist's Perspective. AB - Masses of the head and neck comprise a variety of benign and malignant tumors and tumor-like conditions, which may present diagnostic challenges to the surgical pathologist and surgeon. Fine needle aspiration cytology is an increasingly popular technique in the initial evaluation of such lesions. The high diagnostic accuracy of this technique makes it generally preferable to traditional surgical biopsy. It is particularly useful in the sampling of histologically uniform neoplasms of the salivary glands, identification of classical papillary carcinoma and medullary carcinoma of the thyroid, separation of colloid goiter from follicular neoplasms, and confirmation of clinically suspicious lymph node metastases in cases with already documented malignant diseases. Despite its usefulness, aspiration cytology of head and neck lesions has certain inherent pitfalls, and these are briefly examined in this review. Int J Surg Pathol 8(1):17-28, 2000 PMID- 11493962 TI - Differential Expression of Myoepithelial Markers in Salivary, Sweat and Mammary Glands. AB - Myoepithelial cells (MECs) are contractile elements showing a combined epithelial and smooth muscle phenotype. Among the numerous immunohistochemical markers employed to detect MECs, smooth muscle actin (SMA) is the most widely used. Recently, other markers of smooth muscle differentiation have been demonstrated in MECs, such as calponin, heavy caldesmon (h-caldesmon), and smooth muscle myosin heavy chain (SMM-HC). In the present study normal salivary, mammary, and sweat glands have been studied with four markers of smooth muscle differentiation (SMA, calponin, h-caldesmon, and SMM-HC). The four markers were differentially expressed in the various types of glands. In parotid glands MECs mainly expressed calponin and caldesmon; in submandibular and in cutaneous apocrine and eccrine glands, MECs strongly expressed SMA, calponin, and caldesmon; in minor salivary glands all four markers were equally strongly expressed; and in mammary glands SMA, calponin, and SMM-HC were present both in periductal and periacinar MECs while caldesmon was present in periductal MECs only. In addition to MECs, SMA stained stromal myofibroblasts, sometimes hampering the identification of MECs. Among the other markers, calponin stained only rare stromal myofibroblasts, while caldesmon and SMM-HC were confined to MECs. In conclusion, these latter markers are very useful for identifying MECs. It is suggested that the differential expression of smooth muscle contractile proteins might reflect different functions of MECs in the various sites. Int J Surg Pathol 8(1):29-37, 2000 PMID- 11493963 TI - Annual Rhythm in the Growth of Human Breast Carcinomas as Reflected in the Histology of Their Growing Edge. AB - This histologic study of breast carcinoma, based on 100 consecutive cases, identifies a dynamic scenario at the tumor edge. Three main types of tumor edge were identified. An inactive edge, seen throughout the year, consisted of tumor cells lying in fibrous tissue that merged into the adjacent fatty tissue. An infiltrative edge was characterized by an acute microvascular response and the presence of fine lymph channels often containing embolic tumor cells. A capsular edge showed sharp tumor demarcation accompanied by proliferation and followed by lymphocyte infiltration. Infiltrative edges were found mainly in the first half of the year, and capsular edges in the second. Thus, the growth of individual breast carcinomas appears to be the result of a common series of growth phases at their advancing edge that follow an annual cycle. Int J Surg Pathol 8(1):39-47, 2000 PMID- 11493964 TI - Malignant Melanoma Metastatic to the Testis: A Report of Three Cases with Clinically Significant Manifestations. AB - Three cases of malignant melanoma metastatic to the testis, each of which was a challenge in pathologic interpretation, are reported. The patients were 43, 55, and 80 years of age, respectively. A history of malignant melanoma was not known to the initial examining pathologist in two of the cases. The testicular tumors were all unilateral. One took the form of multiple nodules, but the others were discrete solitary masses. On microscopic examination diffuse, nested, follicular, and fascicular patterns, usually in combination, were seen. The tumor cells had moderate to abundant eosinophilic cytoplasm in two cases, but scanty cytoplasm in a third. In one case there was also a component of cells with copious foamy cytoplasm. Rare areas of melanin pigment were identified in two tumors. Immunohistochemistry for S-100 and HMB-45 was positive in the two cases in which it was performed. In the third case electron microscopy revealed aberrant melanosomes within the tumor cells. The differential diagnosis in these and rare other cases of malignant melanoma involving the testis during life is broad and includes seminoma, malignant lymphoma, and Leydig cell tumor. When the diagnosis is entertained, a search for melanin pigment, immunohistochemical stains for S 100 protein and HMB 45, and further inquiry into the clinical history are often crucial. Int J Surg Pathol 8(1):49-57 2000 PMID- 11493965 TI - Epithelial Phenotype in Ewing's Sarcoma/Primitive Neuroectodermal Tumor. AB - Neural differentiation is an integral component of Ewing's sarcoma/primitive neuroectodermal tumor (PNET), which exhibits a continuous spectrum from minimal to prominent neural phenotype. Differentiation of Ewing's sarcomas/PNETs along other lineages or the expression of an epithelial phenotype is less common and-if present-may cause diagnostic difficulties. In this study we evaluated the frequency of epithelial differentiation in formalin-fixed and paraffin-embedded tissues of 33 (22 primary and 11 metastatic) Ewing's sarcomas/PNETs by using an immunohistochemical assay with several antikeratin antibodies. Focal positivity for low- or high-molecular-weight keratins was documented in 18% of the cases, and diffuse coexpression of low- and high-molecular-weight keratins was observed in two cases. Expression of the MIC-2 gene product was documented in 94% of the tumors. The primitive neural phenotype as revealed by expression of either neuron specific enolase or synaptophysin was observed in 30% of the cases, but coexpression of both neural markers was present in only 15% of the tumors. This study documents that, in addition to primitive neural differentiation, Ewing's sarcomas/PNETs frequently exhibit focal positivity for keratins, with rare strong diffuse coexpression of both low- and high-molecular-weight keratins. The findings indicate that the expression of an epithelial phenotype, at least in a focal fashion, is a relatively frequent finding in otherwise typical Ewing's sarcomas/PNETs. Int J Surg Pathol 8(1):59-65, 2000 PMID- 11493967 TI - Hyaline Globules in Adenoid Cystic Carcinoma. PMID- 11493966 TI - Pleomorphic Angiomyolipoma of Digestive Tract: A Heretofore Unrecognized Entity. AB - Two cases of pleomorphic angiomyolipoma (AML) composed of bizarre epithelioid smooth muscle cells located in the gastrointestinal tract are reported. One involved the appendix of a 6-year-old girl, and the second the cecum of a 22-year old woman. In both instances the tumor cells were immunoreactive for HMB-45 and A103. To the best of our knowledge, this peculiar variant of an AML has not previously been recognized in this location. Int J Surg Pathol 8(1):67-72, 2000 PMID- 11493968 TI - Epithelioid Hemangioendothelioma of the Spermatic Cord. AB - This report describes an epithelioid hemangioendothelioma arising on the spermatic cord of a 50-year-old man who presented with a nodule in the right hemiscrotum. Histologic examination showed a cellular tumor composed of epithelioid cells, many of which had intracytoplasmic lumina containing red blood cells. Immunohistochemistry showed focal positivity of tumor cells with the endothelial markers CD34, factor VIII-related antigen, and Ulex and with the anticytokeratin antibodies CAM5.2 and AE1/AE3. Ultrastructural examination revealed polygonal tumor cells forming vascular spaces and intracytoplasmic lumina containing red blood cells. This is the second documented occurrence of epithelioid hemangioendothelioma at this site. The case illustrates the value of immunohistochemistry and electron microscopy in establishing a diagnosis. Int J Surg Pathol 8(1):75-78, 2000 PMID- 11493970 TI - From Other Sources. PMID- 11493969 TI - Angiomyofibroblastoma of the Male Perineum: An Unusual Location for a Rare Lesion. AB - We report a case of angiomyofibroblastoma (AMF)-like tumor of the perineum in a 54-year-old man. The asymptomatic lesion measured 3 cm and appeared as a tan homogeneous mass. Histologically, it appeared as a circumscribed nodular spindle cell proliferation with alternating cellular and hypocellular areas. The spindle cells exhibited minimal nuclear pleomorphism and scanty mitotic activity. Focally, some cells were epithelioid. Large blood vessels were present, with perivascular fibrosis. The spindle cells expressed vimentin, but not desmin, actin, S100, or CD34. These features were similar to those of AMF as initially reported in the vulva. A perineal localion of this lesion in a male has not been reported in the literature, to the best of our knowledge. Int J Surg Pathol 8(1):79-82, 2000 PMID- 11493972 TI - A Search for the Ideal Fixative. PMID- 11493971 TI - A New Society and Granting Opportunity for Tissue Repositories. PMID- 11493973 TI - A Practical Approach to the Differential Diagnosis of Small Round Cell Tumors of Infancy Using Recent Scientific and Technical Advances. AB - Scientific and technical advances have direct impact on the day to day practice of surgical pathology. Pathologists must learn to use new findings to improve diagnostic classification and add clinically useful information in a fiscally responsible manner. The undifferentiated tumors of young people (historically referred to as small round cell tumors of infancy) are a useful example of how new information has become an important part of routine practice. This article describes a practical approach to the differential diagnosis of small round cell tumors using immunophenotypic and genetic analysis as an aid to increase confidence and diagnostic accuracy. Int J Surg Pathol 8(2):87-97, 2000 PMID- 11493974 TI - Fibrous Lesions of the Breast: A Review. AB - Fibrous lesions of the breast are challenging for a number of reasons. They occur infrequently and there is significant clinical, radiologic, and histologic overlap between reactive and neoplastic lesions, as well as with some nonfibrous lesions. Pure fibrous lesions of the breast encompass reactive conditions, such as scars and nodular fasciitis, and neoplastic entities, such as the distinct benign mammary myofibroblastoma, locally aggressive fibromatosis, and rare true sarcomas. In this review, we focus on nodular fasciitis, mammary myofibroblastoma, and primary mammary fibromatosis. We also briefly discuss spindle cell metaplastic carcinoma because it represents the most important differential diagnosis through its ability to closely simulate nodular fasciitis and fibromatosis. Caution should be exercised in the interpretation of fibrous lesions of the breast, especially if the tissue sample is limited. Int J Surg Pathol 8(2):99-108, 2000 PMID- 11493975 TI - Hu Immunolabeling as a Marker of Neural and Neuroendocrine Differentiation in Normal and Neoplastic Human Tissues: Assessment Using a Recombinant Anti-Hu Fab Fragment. AB - The recombinant antibody fragment Fab GLN 495 recognizes an epitope shared by members of the neuron-associated Hu protein family (including HuC, HuD, and HelNl). This novel reagent labels the nuclei of neurons throughout the peripheral and central neuraxes and has been shown to recognize pulmonary small cell carcinomas and central nervous system (CNS) tumors of mature neuronal phenotype or neuronogenic differentiating capacity. Using this Fab fragment, we have undertaken a systematic survey of normal human tissues and an assessment of 554 non-CNS tumor samples for immunohistochemical evidence of Hu expression. Adrenomedullary cells, pancreatic islet cells, paraganglial chief cells, isolated adenohypophyseal cells, and spermatogonia were the only nonneuronal normal tissue elements to bind Fab GLN 495. In addition to labeling all 10 small cell carcinomas studied (six of which were extrapulmonary in origin), this recombinant anti-Hu Fab proved immunoreactive with neuroblastomas (four/four), esthesioneuroblastomas (one/one), typical (three/four) and atypical (one/four) pulmonary carcinoids, pancreatic islet cell tumors (two/six), large-cell neuroendocrine carcinoma of lung (one/four), Merkel cell tumors (two/three), medullary carcinomas of the thyroid (four/six), pheochromocytomas (two/four) and paragangliomas (four/four). Nonneural/neuroendocrine tumor labeling was restricted to the neuronal and immature neuroepithelial components of teratomas, to extraskeletal myxoid chondrosarcomas (three/four) and to small subsets of cells within examples of renal rhabdoid tumor (one/four), desmoplastic small cell tumor (one/four), alveolar rhabdomyosarcoma (two/four), Ewing sarcoma/PNET (two/nine), and Wilms tumor (one/four). Immunoreactivity was principally nuclear, with variable cytoplasmic labeling. Our findings support the largely restricted expression of Hu by neural/neuroendocrine neoplasms, suggest a potential role for Fab GLN 495 in the identification of small cell carcinomas irrespective of primary site, and support a recent proposal that at least some extraskeletal myxoid "chondrosarcomas" actually represent neuroendocrine tumors of soft parts. Int J Surg Pathol 8(2):109-117, 2000 PMID- 11493976 TI - Diagnostic Value of Calretinin in Mast Cell Lesions of the Skin. AB - The diagnosis of mast cell lesions of the skin can occasionally be challenging. Calretinin, a 29 kD neuron-specific calcium-binding protein found mostly in the CNS and retina, has been shown to be a positive marker for mesotheliomas, and is also expressed in mast cells. We studied the diagnostic value of calretinin and compared our results to other established ancillary studies used to identify mast cells, such as Toluidine blue and the Leder stain. Sixty-three cases were studied, including 45 mast cell lesions (22 urticaria pigmentosum, 17 mastocytomas, and six telangiectasia macularis eruptiva perstans [TMEP]), seven nevi, three melanomas, four granular cell minors of the skin, three cutaneous lymphomas, and one granulocytic sarcoma. Patients ranged in age from less than 1 to 85 years with a median age of 29 years. The group consisted of 36 females and 27 males. Calretinin was expressed in all 45 mast cell lesions. Negative staining for calretinin was seen in all skin lesions that potentially could be considered in the differential diagnosis of mast cell lesions such as nevi, melanomas, lymphomas, and the granulocytic sarcoma. However, calretinin expression was noted in four/four granular cell tumors. Leder and Toluidine blue stains were positive in all 45 mast cell lesions, and all nonmast cell lesions were negative with these stains. In conclusion, our study demonstrated that calretinin is a sensitive and specific marker of mast cells and can be an aid in distinguishing mast cell lesions from other skin lesions considered in the differential diagnosis. Calretinin may be more sensitive than the currently used special stains utilized to diagnose mast cell lesions having few diagnostic mast cells such as TMEP. However, this immunoperoxidase stain does not add significant diagnostic information in most cases, when compared with the currently used less expensive special stains and, therefore, is not cost-effective. Int J Surg Pathol 8(2):119-122, 2000 PMID- 11493977 TI - The Incidence of Apoptosis During Colorectal Tumorigenesis. AB - Epithelial homeostasis in colorectal tumorigenesis is dependent not only on the rate of cell production but also on the rate of apoptosis, a genetically programmed process of autonomous cell death. Ideally, an analysis of cell kinetics should be carried out for both cell proliferation and death. We investigated the incidence of apoptosis in 35 colorectal neoplasms (15 adenomas and 20 carcinomas) using the DNA nick end labeling method (TUNEL). The expression of Ki-67 as a marker of proliferating activity and some kinds of oncogene products were analyzed immunohistochemically. When the TUNEL labeling index (TI) and the Ki-67 labeling index (KI) were determined, TI was found to be significantly higher in adenomas with high-grade dysplasia (TI: 2.5%) than in adenomas with low-grade dysplasia (TI: 0.6%) or carcinomas (TI: 1.4%). In contrast, KI increased with the progression of colorectal tumorigenesis. Moreover, TI of the carcinomas was significantly higher in c-Myc-positive cases than in c-Myc-negative cases (p<0.05). The results indicate that apoptosis plays an important role in the early stage of the adenoma-carcinoma sequence, permitting us to speculate that the increased tumor cell proliferation is negated by increased apoptosis at the stage of adenoma with high-grade dysplasia (or intramucosal carcinoma), while cell proliferation overwhelms cell death at the stage of invasive carcinoma. Int J Surg Pathol 8(2):123-132, 2000 PMID- 11493978 TI - Predicting Metastatic Risk of Gastrointestinal Stromal Tumors: Role of Cell Proliferation and Cell Cycle Regulatory Proteins. AB - Gastrointestinal stromal tumors (GIST) are a heterogeneous group of neoplasms whose biologic behavior is difficult to predict. The aim of this study is to evaluate the prognostic value in GIST of some oncoproteins involved in regulation of cell proliferation. Tumor size, mitosis, necrosis, and p53, c-myc, and bcl-2 protein expression of 32 GIST were studied. Proliferative index was assessed with Ki67. The 32 cases were grouped into the following clinical categories: (1) clinically benign (BN) were defined as disease-free survival greater than 3 years (n=10); (2) clinically malignant (MN) in which local recurrence or metastasis occurred regardless of the follow-up time (n=15); and (3) clinically indeterminate (ID) owing to follow-up <3 years without metastasis or local recurrence (n=seven). Discriminant analysis was used to allocate any tumor to one of the two prognostic groups (BN or MN). In univariate analysis all six factors studied above proved to be of significant prognostic value. Using a multivariate stepwise discriminant analysis to take into account the interrelationship between factors, we found that c-myc expression was the most important prognostic factor, followed, in order of statistical weight, by size and Ki67. These were combined to define a discriminant score ([10.75 x c-myc]+[0.39 x size]+[0.078 x Ki67] 15.54=score), which was capable of correctly identifying tumors in our series whose known clinical behavior was BN or MN in 92% of the cases. The classification score was applied subsequently to the seven clinically ID cases: Three (42.9%) were predicted as BN, and four (57.1%) were predicted as MN. Both expression of oncoprotein c-myc and the proliferative index provide prognostic information in GIST, in addition to morphologically established prognostic factors such as size. These factors in a discriminant analysis proved to be useful for the clinical classification of GIST into BN or MN and to predict the clinical outcome of clinically ID tumors. Int J Surg Pathol 8(2):133-144, 2000 PMID- 11493979 TI - Extraskeletal Myxoid Chondrosarcoma with Rhabdoid Features, with Special Reference to Its Aggressive Behavior. AB - The presence of rhabdoid cells has been reported in various types of malignant neoplasms. Thirty-six cases of extraskeletal myxoid chondrosarcoma (EMC) were reviewed, and three of them revealed rhabdoid features. These cases contained cells having prominent eosinophilic globular cytoplasm, but were otherwise typical of EMC. Immunohistochemically, cytokeratin (CAM 5.2) was positive in three of 20 cases (15%), including two of the three with rhabdoid features. The rhabdoid cells were also immunostained with vimentin. The 5-year survival rate in the 26 cases with follow-up information was 73%, and the 10-year survival rate was 63%. The cases with rhabdoid features had a significantly poorer prognosis (p=0.0271). This study suggests that presence of rhabdoid features is a predictor of aggressive behavior in EMC, as it was shown to be in other mesenchymal neoplasms. Int J Surg Pathol 8(2):145-152, 2000 PMID- 11493980 TI - The Perivascular Cuff of Large Lymphoid Cells: A Clue to Diagnosis of Anaplastic Large Cell Lymphoma. AB - The small cell and lymphohistiocytic variants of anaplastic large cell lymphoma are commonly misdiagnosed as reactive lymphadenopathies. A helpful histologic clue is the presence of perivascular cuffs of large lymphoid cells, and the diagnosis can be readily confirmed by immunostaining for CD30 and ALK. Int J Surg Pathol 8(2):153-156, 2000 PMID- 11493981 TI - A Mistake I Made from Which I Learned Much. PMID- 11493982 TI - Diagnostic Errors in Surgical Pathology Uncovered by a Review of Malpractice Claims: Part I. General Considerations. PMID- 11493983 TI - Meigs' Syndrome: Dr. Richard Cabot's Hidden First American Case. AB - The syndrome known most widely as "Meigs' syndrome" is so named as a result of the description of it by Dr. Joe V. Meigs (a gynecologist with a strong interest in pathology) and Dr. John W. Cass (an internist with a major interest in pulmonary disease) in 1937. In his final writings on the topic 17 years later, Dr. Meigs reviewed the literature prior to his first report in more detail than he had initially and credited appropriately those who had noted the association of ovarian fibromatous tumors, ascites and pleural effusion in earlier times. He did not, however, record the description of the association by the great physician, Dr. Richard C. Cabot in 1912 even though they worked in the same hospital and the case in question was one of those that formed the basis of the report of Meigs and Cass. Int J Surg Pathol 8(2):165-168, 2000 PMID- 11493984 TI - Composite Large-Cell Neuroendocrine Carcinoma and Surface Epithelial-Stromal Neoplasm of the Ovary. AB - The clinicopathologic features of two cases of composite large-cell neuroendocrine carcinoma and surface epithelial-stromal neoplasm of the ovary are reported and those of eight previously published cases reviewed. The patients ranged in age from 22 to 77 years (mean, 56 years). The surface epithelial stromal neoplasm was an endometrioid adenocarcinoma in one case, a mucinous cystadenoma in one case, and a mucinous adenocarcinoma in eight cases. The large cell neuroendocrine carcinoma in these tumors may represent dedifferentiation of the neuroendocrine cells present in the surface epithelial-stromal tumor. This composite tumor type is highly aggressive. Of eight patients with follow-up information, all had died of disseminated tumor; six within 10 months, and two in 19 months and 3 years, respectively, after diagnosis. Only the neuroendocrine carcinoma component was found in the metastatic sites. Int J Surg Pathol 8(2):169 174, 2000 PMID- 11493985 TI - Genes and Cancer. PMID- 11493987 TI - Guest Editorial: Two Proposals Regarding the Terminology of Thyroid Tumors. PMID- 11493986 TI - New Entities in Pathology. PMID- 11493988 TI - ret/PTC-1 Activation in Hashimoto Thyroiditis. AB - Activation of ret/PTC-1 has been documented in a minority of papillary thyroid carcinomas (PTC). In a recent study, the authors' group detected the presence of ret/PTC-1 in association with a background of florid lymphocytic thyroiditis (LT) in 58% of cases of PTC studied, which prompted them to examine the incidence of RET/PTC-1 expression in 27 examples of various forms of nonlymphomatous lymphoid infiltration of the thyroid by using TaqMan RT-PCR. Overall, 21 cases (78%) were found to express the chimeric transcript of ret/PTC-1. Eighteen cases of Hashimoto thyroiditis were positive (95%), and, of these, three had concomitant PTC while the remainder had no histologic evidence of associated malignancy. Three cases of lymphocytic thyroiditis demonstrated activated ret/PTC-1 (43%), two having associated PTC. These data suggest either that ret/PTC-1 is an indicator of follicular thyroid cell activation or that ret/PTC-1 activation is an early event in malignant transformation. If the latter is the case, it may be that, in a defined subset of the cell population, ret/PTC-1 activation elicits an autoimmune response, which, while possibly curtailing the development of PTC in the majority of cases, results in destruction of the thyroid parenchyma. Int J Surg Pathol 8(3):185-189, 2000 PMID- 11493989 TI - Adhesion Molecules as Diagnostic Tools in Tumor Pathology. AB - Adhesion molecules are transmembrane glycoproteins mediating cell-cell and cell extracellular matrix interactions. They control a number of fundamental biological processes including cell migration, differentiation, proliferation, and apoptosis. In the last decade there has been an increasing interest in the exploitation of these molecules as diagnostic and/or prognostic markers in tumor pathology. For example, a large number of studies have shown that loss of E cadherin expression correlates with high tumor grade and advanced tumor stage in a number of malignancies. The analysis of adhesion molecule profile in a routine clinical setting needs further investigation in prospective multicenter studies. Int J Surg Pathol 8(3):191-200, 2000 PMID- 11493990 TI - Application of CARD-ISH for Assessment of Numerical Chromosome Aberrations in Interphase Nuclei of Human Tumor Cells. AB - In this study based on the study of a centromeric DNA probe specific for chromosome 8 the authors standardized a method for the enzymatic detection of specific chromosome copy numbers on interphase nuclei from tumor tissue samples. Since the in situ hybridization (ISH) of chromosome 8 specific probe was revealed with a catalyzed reporter deposition (CARD), which allows a high amplification of the hybridization signal, the method was designated as CARD-ISH. This method has been standardized on interphase nuclei isolated from clinical samples of pituitary adenomas, as well as on human normal lymphocytes. On the same samples, they also evaluated chromosome 8 copy number distribution by FISH. Comparison between CARD-ISH and FISH results showed no significant differences between the two methods, proposing CARD-ISH as a reliable alternative to FISH for chromosome numerical aberration assessment in laboratories that do not have specific facilities for epifluorescence microscopy or cytogenetics and that need a long term storage of slides that had been used for diagnostic purposes. Int J Surg Pathol 8(3):201-206, 2000 PMID- 11493991 TI - Allelic Deletion of VHL Gene Detected in Papillary Tumors of the Broad Ligament, Epididymis, and Retroperitoneum in von Hippel-Lindau Disease Patients. AB - Although clinically associated with von Hippel-Lindau (VHL) disease, the pathogenesis of papillary tumors of the broad ligament, epididymis, and peritoneum arising in patients with VHL disease is not clear. The "classic" VHL associated neoplasms, including hemangioblastoma and renal ceil carcinoma, have been found to be associated with the inactivation of both VHL gene copies. It is not known whether a similar inactivation of the VHL gene is also responsible for the development of these uncommon VHL-associated lesions. The authors performed PCR (polymerase chain reaction) and PCR-based SSCP (single-strand conformation polymorphism) analysis on five predominantly papillary tumors in five VHL patients (one papillary cystadenoma of the broad ligament, one endometrioid cystadenoma of the broad ligament, two papillary cystadenomas of the epididymis, one papillary tumor of the retroperitoneum) with four polymorphic markers of VHL gene (D3S1038, D3S1110, D3S2452, 104/105). All five tumors showed allelic loss of VHL gene. The results provide the first genetic evidence for the role of VHL gene in the tumorigenesis of these rare benign neoplasms and confirm these tumors as phenotypic manifestations of VHL disease. Int J Surg Pathol 8(3):207-212, 2000 PMID- 11493992 TI - p53 Overexpression and Steroid Hormone Receptor Status in Endometrial Carcinoma. AB - Mutations of the p53 tumor suppressor gene often occur in a variety of human malignant tumors and are frequently associated with overexpression of p53 protein. This study was designed to examine indirectly the frequency of p53 protein in primary endometrial carcinoma and to correlate the overexpression with steroid hormone receptor status including pS2 protein status. The study was performed on 79 formalin-fixed, paraffin-embedded tissues of endometrial carcinoma. P53 protein overexpression was detected by means of immunohistochemistry using monoclonal antibody NCL-p53-DO7. Estrogen and progesterone receptor status was determined by immunohistochemistry using the monoclonal antibodies NCL-ER-LH(2) and NCL-PGR, respectively, and the pS2 protein using polyclonal antibody NCL-pS2. Overexpression of p53 protein was found in 27 (34%) of the 79 endometrial carcinomas. A strong positive relationship was demonstrated between histologic grade and p53 protein overexpression. There was a significant correlation between p53 protein overexpression and negative estrogen receptor status (49%) negative progesterone receptor status (49%) as well as a negative pS2 protein (45%). The results suggest that overexpression of p53 is associated with high malignant potential. However, p53 overexpression itself does not appear to be an independent prognostic factor in endometrial carcinomas. Int J Surg Pathol 8(3):213-222, 2000 PMID- 11493993 TI - Two Previously Unemphasized Features of Endometriosis: Micronodular Stromal Endometriosis and Endometriosis with Stromal Elastosis. AB - This report draws attention to two unusual features of the stromal component of endometriosis that can create problems in diagnosis. One of these is the exclusive presence of endometriotic stroma, so-called stromal endometriosis, that can occasionally take the form of microscopic nodules of endometriotic stroma on the pelvic peritoneum and other locations. This finding, which we refer to as a micronodular stromal endometriosis, can be overlooked entirely or lead to confusion with peritoneal involvement by low-grade endometrial stromal sarcoma. The other finding we describe is the presence of a prominent elastotic stromal response to endometriosis that can occasionally obscure or even focally obliterate the typical endometriotic stroma. This finding, although nonspecific, can be a diagnostic clue to the presence of endometriosis, especially when accompanied by the presence of the typical endometriod glands of that lesion. Int J Surg Pathol 8(3):223-227, 2000 PMID- 11493994 TI - Diagnostic Pitfalls in Surgical Pathology- Uncovered by a Review of Malpractice Claims: Part II. Breast Fine Needle Aspirations. AB - Breast fine needle aspiration (FNA) biopsy accounted for 6% of surgical pathology and FNA claims reviewed from 1995-97. The majority of these claims were for false negative breast FNA resulting from sampling error in a woman with a palpable breast mass. Controversies concerning the definition of specimen adequacy for breast FNA are discussed and a strategy for minimizing the management and liability consequences of the problem is proposed. Claims for false positive breast FNA usually result from interpretive error. Int J Surg Pathol 8(3):229 231, 2000 PMID- 11493995 TI - The Prolonged Gestation, Birth, and Early Life of the Sex Cord Tumor with Annular Tubules and How it Joined a Syndrome. AB - The sex cord tumor with annular tubules was discovered as a distinctive entity when its unusual-appearing microscopic pattern was encountered repetitively in a pathology consultation practice. The author was stimulated to describe the clinicopathological features of the tumor and assign it a specific name when a minority of the cases in his series were found to be associated with the rare Peutz-Jeghers syndrome. Both these tumors and those occurring in the absence of the syndrome have been shown to have distinctive clinicopathological features that differ from those of other tumors in the sex cord-stromal category. Int J Surg Pathol 8(3):233-238, 2000 PMID- 11493997 TI - Pleuropulmonary blastoma type 1, specimen partially deroofed and photographed under water. PMID- 11493996 TI - Signet ring cells in ischemic ileitis. PMID- 11493998 TI - Primary Adenoidal Hodgkin's Disease: Report of a Case with an Unusual Morphology and Review of the Literature. AB - The nasopharyngeal lymphoid tissue (adenoids) is an uncommonly reported primary site for Hodgkin's disease. We report a case of primary adenoidal, interfollicular, epithelioid cell-rich variant of mixed cellularity Hodgkin's disease. The combination of an interfollicular pattern and richness of epithelioid histiocytes made it very difficult to make the diagnosis of Hodgkin's disease without the help of immunohistochemistry. To our knowledge, only 39 cases of Hodgkin's disease primarily involving the Waldeyer's ring have been reported in the English literature, 24 of these primarily involving the adenoids. Our case shows the difficulty encountered in making the diagnosis when a very unusual morphologic appearance of Hodgkin's disease is seen at a rare presentation site. The appropriate immunohistochemical work-up should be performed in unusual lymphohistiocytic proliferations involving the Waldeyer's ring. Int J Surg Pathol 8(3):241-246, 2000 PMID- 11493999 TI - Melanotic Neuroectodermal Tumor of Infancy: Clinicopathologic Study of a Case, with Emphasis on the Chemotherapeutic Effects. AB - A case of melanotic neuroectodermal tumor of infancy (MNTI) in a 6-months-old white female child is presented. The tumor was located in the left half of the superior maxillary bone and affected the maxillary sinus, inducing large facial asymmetry. The histologic aspects of the tumor were typical for the entity: two cell populations, one pigmented melanocyte-like cell, the other small neuroblastic-like cell. After adjuvant chemotherapeutic treatment with a strong reduction of the tumor, the melanocyte-like cell predominated, allowing a better result of the surgery and probably a better prognosis. Int J Surg Pathol 8(3):247 251, 2000 PMID- 11494000 TI - Warty Carcinoma Arising in Condyloma Acuminatum of Urinary Bladder: A Case Report. AB - We describe the case of a 62-year-old man with chronic irritation of the urinary bladder resulting in dysuria and hypogastric pain. Three neoplasms measuring 0.5, 1, and 1.5 cm, respectively, were observed by cystoscopy and removed by transurethral resection (TUR). Histologic examination showed a complex folding of squamous hyperplastic epithelium around a connective tissue core. The superficial epithelium contained numerous koilocytes. The double polymerase chain reaction (PCR) detected DNA of type 11 human papillomavirus (HPV). The diagnosis was condyloma acuminatum of bladder. Three months later the patient presented with fever, and a new cytoscopy demonstrated an ulcerated, exophytic 4.5 cm mass. Histopathology showed a squamous carcinoma with papillomatous structure, pronounced viral koilocytosis, and irregular invasive deep margin. HPV type 11 was found with double PCR. The diagnosis was warty carcinoma arising in condyloma acuminatum. To the best of our knowledge, this is the first case of warty carcinoma of the urinary bladder described in the literature. We discuss the relationship between the infection by HPV and the development of condyloma acuminatum, its evolution toward a well-differentiated squamous carcinoma, and its distinction from verrucous carcinoma. Int J Surg Pathol 8(3):253-259, 2000 PMID- 11494002 TI - Post-Chernobyl International Cooperation on Thyroid Tumors. PMID- 11494001 TI - Digital Pathology: Science Fiction? PMID- 11494003 TI - K-ras Gene Mutations in Liver Carcinomas from a Mediterranean Area of Spain. AB - The prevalence of human liver cancer shows a broad spectrum of variation in distinct geographical regions, depending on different risk factors, and their pathogenesis is poorly understood. We study the significance of molecular alterations of K-ras gene in human liver carcinomas in a low-incidence region such as a Mediterranean area of Spain (Valencia and Gerona). Our results reveal a low incidence of K-ras codon 12 mutations in hepatocellular carcinomas (HCC) and a higher incidence in cholangiocarcinomas (CCC). Similar results have been obtained in high-risk areas. We conclude that K-ras gene mutations are not a major event in the malignant transformation of hepatic cells in this region of the Mediterranean but that molecular implications of hepatocellular and cholangiocarcinomas appear to be different. Int J Surg Pathol 8(4):267-270, 2000 PMID- 11494004 TI - Differential Diagnosis Between Intraductal Papillary-Mucinous Tumors and Mucinous Cystic Tumors of the Pancreas. AB - Mucinous cystic tumors (MCTs) and intraductal papillary-mucinous tumors (IPMTs) are often confused with each other. However, clinicopathological studies have shown that these are two distinct entities. In this review, clinicopathological differences and important features for differential diagnosis are presented. MCTs are cyst-forming tumors in the body or tail of the pancreas seen almost exclusively in females. IPMTs show distinct duct ectasia in the pancreatic head with male predominance. Histologically both tumors consist of mucin secreting tall columnar epithelium. MCTs are often accompanied by characteristic "ovarian type stroma." Recent immunohistochemical and gene analysis have not demonstrated significant differences between two tumor types. These tumors should be clearly separated from ordinary ductal adenocarcinomas of the pancreas as they follow indolent course. Int J Surg Pathol 8(4):271-278, 2000 PMID- 11494005 TI - Uterine Endometrioid Carcinoma with Small Nonvillous Papillae: An Analysis of 26 Cases of a Favorable-Prognosis Tumor To Be Distinguished from Serous Carcinoma. AB - We have encountered a number of endometrioid carcinomas with small papillary buds lacking fibrovascular cores that could be confused with the small cellular papillae of serous papillary carcinoma (SPC). We have designated these tumors "endometrioid carcinoma with small nonvillous papillae" (ECSP). Because they have not been investigated previously we analyzed 26 examples and compared their features with those of 21 SPCs of the uterus. Three hundred and ninety consecutive cases of endometrial carcinoma diagnosed between January, 1989, and January, 1994, were retrieved from our hospital files; 26 (6.7%) of them, (8% of the endometrioid carcinomas) were identified as ECSP, and 21 (5.4%) as SPC. Tumors were classified as ECSP when the small papillae were present within the glands of otherwise typical endometrioid carcinoma or on the villous projections of villoglandular endometrioid carcinoma. Most of the papillae were in the form of buds of cells with ample eosinophilic cytoplasm and a low nuclear-to cytoplasmic ratio, but some papillae had a more complex pattern. The papillae arose on a background of International Federation of Gynecology and Obstetrics (FIGO) grade 1 or 2 endometrioid carcinomas, which showed squamous differentiation in half the cases. SPCs were identified according to generally accepted criteria. The mean age of the patients with ECSP was 67 years, intermediate between that of the patients with endometrioid carcinoma lacking small nonvillous papillae (62 years) and that of the patients with SPC (71 years). Patients with ECSP more frequently presented at an earlier stage (73% stage I/II) than those with SPC (29% stage I/II). The overall 5-year survival of patients with ECSP was 84% (95%CI: 0.68-1), more than double that of patients with SPC, 33% (95%CI: 0.10-0.56). ECSP may be confused with SPC on microscopic examination but has clinical and pathological features similar to those of endometrioid carcinoma lacking small nonvillous papillae, and unlike SPC, should be treated in the same manner as the former. Int J Surg Pathol 8(4):279-289, 2000 PMID- 11494006 TI - Mesenchymal Chondrosarcoma: A Small Cell Neoplasm with Polyphenotypic Differentiation. AB - We analyzed the clinicoradiographic, microscopic, and immunophenotypic features of 21 tumors from 13 patients with mesenchymal chondrosarcoma (11 primary and 10 metastatic) and addressed the issue of their potential polyphenotypic differentiation. The immunophenotypic profile of the tumors was analyzed by studying the expressions of the MIC2 gene protein (p30/32(MIC2)), S-100 protein, desmin, myoD1, myogenin, myoglobin, smooth-muscle actin, cytokeratin, neuron specific enolase, and HMB-45. The expression of p30/32(MIC2) was typically restricted to the small cell component of the mesenchymal chondrosarcoma and could be documented in 17 tumors. The cartilaginous areas were positive for S-100 protein in 20 tumors. Scattered positivity of small cells for desmin was seen in 8 cases. In 2 primary tumors from different patients (1 intraosseous and 1 extraskeletal) a diffuse expression of desmin with focal coexpression of myoD1 was present within the small cell component of the tumor. The positivity for smooth-muscle actin was documented in 2 cases. Either the small cell or cartilaginous components were at least focally positive for neuron-specific enolase in 11 tumors. All tumors were negative for myogenin, myoglobin, cytokeratins (AE1/AE3, CAM5.2) and HMB-45. This study showed that, in addition to cartilaginous differentiation, mesenchymal chondrosarcomas may exhibit focal expression of desmin. In rare cases more diffuse rhabdomyoblastic differentiation can be seen within the small cell component of the tumor. Thus, mesenchymal chondrosarcoma is another primitive neoplasm with polyphenotypic differentiation and features that overlap those of other small cell malignances of bone and soft tissue. Int J Surg Pathol 8(4):291-301, 2000 PMID- 11494007 TI - Carcinoma Arising in Microglandular Adenosis: An Immunohistochemical Analysis of 20 Intraepithelial and Invasive Neoplasms. AB - Microglandular adenosis (MGA) of the breast is an uncommon, benign lesion that may mimic invasive carcinoma and has recently been recognized as having significant premalignant potential. When carcinomas arise in MGA, there is often a transition from ordinary MGA to atypical MGA (AMGA) to carcinoma. Nineteen cases of carcinoma arising in MGA are reported: 7 invasive carcinomas, 7 intraductal carcinomas (DCIS), and 5 with both invasive and intraductal carcinoma. A single case of AMGA without carcinoma is also reported. The 20 patients ranged in age from 36 to 81 years (mean 52). The most common clinical presentation was either a palpable mass (13 patients) or a mammographic abnormality (4 patients). All 20 cases contained AMGA, and in some cases AMGA was the predominant lesion. In 18 of the 19 cases with carcinoma, there was a clear transition from AMGA to the carcinoma. Twelve cases contained ordinary MGA, but in only 2 cases was MGA a prominent component of the lesion. In contrast to ordinary MGA, the glands of AMGA were more irregularly shaped, closely packed, and cytologically atypical and tended to lack secretions. A solid, occlusive proliferation of cells in the tubules was seen in 10 cases. All 12 examples of in situ carcinoma were either grade 2 or 3 and typically showed a solid proliferation of severely atypical cells within the glands; a cribrifrom pattern was also present in 1 case. The invasive carcinomas were morphologically diverse and included 2 with a basaloid morphology and 2 metaplastic carcinomas. Various immunostains were performed, and each lesion (AMGA, in situ, and invasive carcinoma) was separately assessed for immunoreactivity. As expected, S-100 was positive in the vast majority of AMGA and in situ carcinomas and in all 12 invasive carcinomas. S-100beta was also positive in the majority of cases although the staining was weaker. Laminin and type IV collagen highlighted the basement membrane around the AMGA and in situ carcinoma and are useful stains in difficult cases. Except for a single case, ER and PR were negative in all lesions. Cytokeratin 7 (CK 7) was positive, while cytokeratin 20 (CK 20) was negative in all cases. Immunostains for CK903 showed no reactivity in any of the invasive carcinomas, in situ carcinomas, or atypical MGA but was focally present in the associated MGA in 2 of the 8 cases studied. Immunostains for MIB-1 and p53 were semiquantitatively assessed and both were positive in AMGA but tended to show a more intense staining in the carcinomas. Five cases were also studied for immunoexpression of alpha-1 antitrypsin (AAT), alpha-1 antichymotrypsin (ACTP), lysozyme, and salivary gland amylase. All 5 invasive carcinomas were positive for ACTP, though the staining was very focal in about 10% of the cells in a basaloid carcinoma. The in situ carcinoma as well as the AMGA in 4 of the 5 cases were positive for ACTP. Three of the 5 invasive carcinomas were positive for AAT in 10% to 40% of the cells. The most intense positivity for AAT and ACTP was in cells with coarsely granular apocrine appearance evident in 2 of the 5 cases. Four of the 5 invasive carcinomas were positive for lysozyme in 10% to 50% of the cancer cells; the in situ carcinoma and the associated AMGA showed similar immunoreaction in each case. None of the 5 cases showed convincing positivity for salivary gland amylase. The MGA in all 5 cases was negative for AAT and ACTP; the MGA in 1 of the 5 cases was positive for lysozyme. This study confirms the potential of MGA to develop into an invasive carcinoma, more clearly defines the features of AMGA, highlights the importance of AMGA in the evolution of carcinoma from MGA, and expands our knowledge of the immunophenotype of AMGA and the carcinomas arising from it. The diagnostic criteria briefly noted previously for diagnosis of AMGA and carcinoma arising in MGA are expanded and formally proposed. Int J Surg Pathol 8(4):303-315, 2000 PMID- 11494008 TI - 34betaE12 Cytokeratin Immunodetection in the Differential Diagnosis of Small Cell Tumors of Lung. AB - The group of small cell tumors of the lung includes fine following: (1) small cell carcinoma (SCC) of neuroendocrine (NE) origin, (2) poorly differentiated squamous carcinoma, (3) the rare basaloid (basal cell) carcinomas, and (4) malignant lymphomas, primitive neuroectodermal tumors (PNETs), and rhabdomyosarcomas. The differential diagnosis among these entities carries a heavy therapeutic impact but may be difficult in small biopsy specimens or in cytologic material, especially if necrosis or artifactual alterations are present. The use of additional techniques such as immunostaining for NE markers is not always helpful, since immunoreactive chromogranin A is detectable in only a small percentage of small cell carcinomas. It has recently been reported that in the aerodigestive tract 34betaE12 cytokeratin (CK) immunostaining selectively labels non-NE carcinomas, including squamous cell carcinoma, adenocarcinoma, and the rare basaloid carcinoma. We evaluated the role of such CK immunodetection in the differential diagnosis of small cell lung tumors in cytologic and biopsy specimens. Eighty-one lung tumors diagnosed by means of endoscopic bronchial biopsy, fine needle aspirate, or bronchial washing were collected. They included 43 small cell NE carcinomas and 38 cases used as controls (comprehensive of 2 large cell neuroendocrine carcinomas, 4 carcinoid tumors, 30 cases of non-NE lung carcinomas, 2 cases of bronchial infiltration by non-Hodgkin lymphomas). 34betaE12 CK immunoreactivity was found in 29/30 cases of non-NE carcinomas, but in only 3/43 SCCs. The latter showed positivity in only a few scattered cells. The 2 cases of bronchial infiltration by malignant lymphoma as well as the 4 cases of carcinoid tumors and the 2 cases of large cell neuroendocrine carcinomas were negative. These findings were confirmed in the surgical specimens of operatedon cases. We conclude that, in lung carcinoma biopsies showing a small cell pattern, presence of 34betaE12 CK immunoreactivity favors a non-NE carcinoma, whereas its absence supports the diagnosis of SCC. Int J Surg Pathol 8(4):317-322, 2000 PMID- 11494009 TI - Hyalinized Uterine Mesenchymal Neoplasms with HMB-45-Positive Epithelioid Cells: Epithelioid Leiomyomas or Angiomyolipomas? Report of Four Cases. AB - We present 4 cases of uterine mesenchymal tumors that were located mostly in the myometrium in middle-aged women. Grossly the tumors vaguely resembled conventional leiomyomas. All tumors were extremely hyalinized. The tumor cells were remarkable because of their large size and epithelioid shape. They occurred singly or more often in variously sized clusters with pericellular clear spaces looking like a halo. The extracellular matrix often formed lacunae around the neoplastic cells. The lacunar type of growth was somewhat reminiscent of immature cartilage. In some places the cells lined up in vague cord-like structures. The epithelioid cells often grew within vessel walls and subendothelially, simulating angioinvasion. In 2 cases there were periodic acid-Schiff (PAS)-positive, needle shaped crystalloids in the cytoplasm of the tumor cells. No atypias and no mitoses were found in any of the 4 tumors. Mucicarmine stain was negative in all cases. Immunohistochemically the tumor cells were negative for cytokeratins, synaptophysin, chromogranin, epithelial membrane antigen (EMA) S-100 protein, and inhibin. They were instead strongly positive for smooth muscle actin, desmin and HMB-45. All patients are disease free after a 1-4-year follow-up. Int J Surg Pathol 8(4):323-328, 2000 PMID- 11494010 TI - Massive Exfoliation of Markedly Atypical Cells in Urine Resulting from Polyoma Virus Interstitial Nephritis. AB - Polyoma virus nephritis is increasingly being recognized as a cause of allograft dysfunction in renal transplant patients. We describe and illustrate the findings in a renal biopsy and a concurrent urine sample from a patient with polyoma virus nephritis. The urine cytology findings were highly suggestive of a tumor due to the presence of large number of atypical cells with marked hyperchromasia, high nuclear/cytoplasmic ratio in a necrotic background. Scattered cells with viral cytopathic changes were also identified, therefore indicating an infectious process with associated reactive cellular changes rather than a tumor. The cytological findings could thus be explained by the viral cytopathic and cytolytic changes, as well as marked reactive and degenerative changes in tubular epithelial cells. Int J Surg Pathol 8(4):329-331, 2000 PMID- 11494011 TI - A Mistake I Made from Which I Learned Much. PMID- 11494013 TI - A Look at an Unusual Animal Kingdom. Part I. Earth's Creatures. PMID- 11494012 TI - Diagnostic Errors in Surgical Pathology Uncovered by a Review of Malpractice Claims. Part III. Breast Biopsies. PMID- 11494014 TI - The "Gourmet" Pathologist. PMID- 11494015 TI - Notes on Collecting Classic Pathology Textbooks. PMID- 11494016 TI - Epstein-Barr Virus-Associated Cholangiocarcinoma with Lymphoepithelioma-Like Component. AB - We report the case of a 19-year-old woman who presented with a hepatic mass without cirrhosis. Light microscopy revealed a cholangiocarcinoma having both well-differentiated adenocarcinoma and lymphoepithelioma-like undifferentiated carcinoma components. By immunohistochemistry, the tumor showed strong and diffuse expression for cytokeratin AE1, 5D3, and CK22. The tumor cells were positive for p53 protein (more than 75% of the cells) but negative for bcl-2 and LMP1. Abundant Epstein-Barr virus EBER (1/2) oligonucleotides were detected in both tumor components, but not in the lymphoid stroma or the nontumor liver. To the best of our knowledge, this is the third report of an Epstein-Barr virus associated primary hepatobiliary adenocarcinoma with lymphoepithelioma-like component. Int J Surg Pathol 8(4):347-351, 2000 PMID- 11494017 TI - Esophageal Plexiform Schwannoma. AB - We report a patient with a plexiform schwannoma located in the cervical part of the esophagus. A large pedunculated intraluminal polyp, originating in the postcricoid region and protruding into the oropharynx, was found in association with multiple submucosal nodules. Both the intraluminal polyp and submucosal nodules consisted of cellular schwannomatous tissue. The patient showed no signs of neurofibromatosis or schwannomatosis. Plexiform schwannomas are uncommon tumors, mainly confined to the dermis and subcutis of trunk, head, and neck region and upper extremities. Visceral examples are vanishingly rare, and a location in the esophagus has not yet been described. The intruiging mode of presentation in the present case, mimicking that of so-called giant esophageal fibrovascular polyp, probably relates to mechanical factors inherent to the unique esophageal location, Int J Surg Pathol 8(4):353-357, 2000 PMID- 11494019 TI - Warty Carcinoma of Bladder Containing HPV Type 11. PMID- 11494018 TI - Vaginal Leiomyoma with Heterologous Paragangliomatous Elements. AB - A vaginal leiomyoma with heterologous paragangliomatous elements is reported. The patient was a 62-year-old woman who presented with uterovaginal prolapse and an asymptomatic vaginal mass that had been present for many years. Histology of the excised mass showed a leiomyoma in a submucosal location, with irregularly shaped islands of chief cells scattered throughout the lesion. These cells were arranged in nests and were surrounded by S100 protein-positive sustentacular cells. The chief cells showed immunoreactivity with chromogranin, synaptophysin, and neuron specific enolase. Although various heterologous elements are commonly encountered within uterine leiomyomata, such elements have not been described within vaginal leiomyomata. Furthermore, the occurrence of paragangliomatous tissue within leiomyomata has not been reported to date. Int J Surg Pathol 8(4):359-365, 2000 PMID- 11494020 TI - Co-expression of beta-adrenergic receptors and cyclooxygenase-2 in pulmonary adenocarcinoma. AB - Pulmonary adenocarcinoma (PAC) is the leading type of lung cancer and is highly resistant to conventional cancer therapy. A better understanding of the regulatory mechanisms which control the growth of this deadly malignancy are urgently needed to develop more effective cancer intervention strategies. Recent studies have shown that PAC frequently overexpresses cyclooxygenase-2 (COX-2). This enzyme converts arachidonic acid (AA) into several metabolites, some of which have been identified as modulators of mitogenesis and apoptosis. Accordingly, the AA cascade and COX-2 are currently widely studied as potential targets for lung cancer prevention. Recent studies by our research group have shown that cell lines derived from human PACs express beta1- and beta2-adrenergic receptors, which regulate the release of AA and DNA synthesis. Moreover, we have demonstrated that an antagonist for beta-adrenergic receptors or aspirin inhibited the development of experimentally induced PAC in a hamster model. These findings suggest that beta-adrenergic receptors may serve as upstream regulators of AA and COX-2-mediated PAC growth. However, no information is currently available on the expression of beta-adrenergic receptors and its possible correlation with the expression of COX-2 in tissue samples from human PAC, casting some doubt on the significance of these findings in vitro and in an animal model. In the current study, we have therefore analyzed tissue samples of human PACs for the expression of beta1-and beta2-adrenergic receptors as well as COX-2 by reverse transcription polymerase chain reaction (RT-PCR) or immunohistochemistry. Our data show that seven out of eight samples co-expressed COX-2 and one or both of these beta-adrenergic receptors, supporting the experimental evidence for a functional link between these neurotransmitter receptors and the AA cascade in the regulation of human PAC. PMID- 11494021 TI - High level amplification of 1p32-33 and 2p22-24 in small cell lung carcinomas. AB - Small cell lung cancer (SCLC) is a frequently occurring, highly aggressive tumor with a generally poor clinical outcome. In order to approach the genetic mechanisms behind the tumor progression, a general screen for DNA copy number alterations was performed using comparative genomic hybridization (CGH). In the series of 23 cases analyzed, CGH alterations were frequently detected ranging from 9 to 22 abnormalities in the individual tumors. The most frequent losses were detected on chromosome arms 3p (23/23), 13q14-21 (23/23), 4p (20/23), 4q (20/23), and 2q22-24 (18/23), while gains preferentially involved chromosome arms 19p (18/23), 19q (17/23), 1p31-35 (15/23), 17q22-25 (11/23), and 5p14-15.3 (9/23). In addition, high level amplification at chromosome arms 1p32-33 and 2p22 24 were found in three and two cases, respectively. Candidate genes for these amplifications include the l-MYC (1p32) and n-MYC (2p24.1) oncogenes, which have been previously found to be overexpressed in SCLC. Taken together, the findings demonstrate a high level of chromosomal instability in SCLC, which is well in agreement with the highly malignant phenotype of this tumor type. Subchromosomal regions involved in gains and losses were delineated and the amplifications of 1p32-33 and 2p22-24 were demonstrated, thus providing starting points for the exact characterization of molecular events involved in SCLC tumor progression. PMID- 11494022 TI - A single physiologic dose of ultraviolet light exposure to human skin in vivo induces phosphorylation of epidermal growth factor receptor. AB - Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein of approximate 180 kDa. EGFR is involved in organ morphogenesis, maintenance and repair of tissues, but its signaling has also been shown to be associated with tumor progression. Our study employing immunostaining technique in vivo human skin demonstrated that physiologic dose of UV (4X MED) exposure to human skin enhanced the constitutive level of EGFR by 2.5+/-0.5-fold at 6 and 24 h after UV exposure in comparison to normal skin but declined at 48 h time point. Basal cell layer predominantly expressed EGFR in comparison to other cellular layers of epidermis and account for about 4.0+/-0.5-fold induction when compared to normal non-UV exposed skin. Constitutive EGFR staining was found all over epidermal layers in normal skin samples. In identical experimental conditions, UV exposure to skin induces phosphorylation of EGFR, which has been confined to matured and differentiated layers of the epidermis. Basal cells are completely devoid of EGFR phosphorylation. UV induction of EGFR phosphorylation was found at peak level at 6 h time point after UV exposure. At 24 h time point it remained elevated but appears diffused and declined, however EGFR phosphorylation was markedly declined at 48 h after UV exposure. Results obtained in immunoperoxidase staining were also confirmed by immunoblot analysis where higher induction of EGFR phosphorylation was observed at 6 h after UV exposure, and marked reduction was found at 48 h time point. Normal skin did not show EGFR phosphorylation. EGFR and its downstream signaling have been shown to be associated with cancer progression and its metastasis, thus blocking the EGFR and its downstream signaling molecules can be employed as the targets for therapeutic intervention against solar UV light induced skin cancer in human population. To our knowledge, this is the first in vivo human study, which clarifies the difference in cellular localization of UV-induced constitutive and phosphorylated forms of EGFR in epidermal cells. PMID- 11494023 TI - PPAR-gamma ligands inhibit growth of human esophageal adenocarcinoma cells through induction of apoptosis, cell cycle arrest and reduction of ornithine decarboxylase activity. AB - Peroxisome proliferator-activated receptor gamma (PPAR-gamma), a member of the nuclear hormone receptor superfamily, is involved in suppression of growth of several types of tumors such as liposarcoma, breast cancer, prostate cancer, and colon cancer, possibly through induction of cell cycle arrest and/or apoptosis. In this study, we demonstrated expression of PPAR-gamma mRNA and protein in human esophageal carcinoma cells. Expression of PPAR-gamma protein was higher in an adenocarcinoma cell line (TE-7 cells) than in a squamous cell carcinoma cell line (TE-1 cells). PPAR-gamma ligands such as 15-deoxy-Delta12,14-prostaglandin J2 and troglitazone significantly inhibited the growth of TE-7 cells but had less or no effect on growth of TE-1 cells. 15d-PGJ2 and troglitazone induced apoptosis in TE 7 cells but not in TE-1 cells. Troglitazone caused G1 cell cycle arrest and reduced ornithine decarboxylase activity (ODC) in TE-7 cells but not in TE-1 cells. Inhibition by PPAR-gamma ligands of growth of esophageal adenocarcinoma cells may thus be due to induction of apoptosis, G1 cell cycle arrest and reduction of ODC activity. PMID- 11494024 TI - Virological and immunological connotations of apoptotic and anti-apoptotic forces in neoplasia. AB - Against the background of its earliest recognition, programmed cell death (PCD) or apoptosis (A) is presented in its fundamental biological contexts. Techniques of its demonstration are listed. Former original works of the authors encompass designs for genetically engineered oncolytic viruses. Presented here are observations on mesenchymal stromal cells of the bone marrow serving as feeder layers to chronic lymphocytic leukemia (CLL) cells (recently rediscovered elsewhere as subverted "nurse cells" protecting CLL cells from A). A-resistant human melanoma cells are shown to expropriate the Fas ligand to Fas receptor (CD95; APO-1) (FasL-->FasR) system for their autocrine growth loop not only in melanoma cells coexpressing CD95 and its ligand but also in CD95-positive melanoma cells undergoing divisions when exposed to CD95 ligand. Bi-directional A induction is demonstrated upon the encounter of cytotoxic lymphocytes and targeted tumor cells as exemplified with lymphomas; and chemotherapy-induced A of malignant cells as exemplified by paclitaxel-induced PCD of Reed-Sternberg (RS) cells in a case of chemotherapy-resistant Hodgkin's disease (HD). A list of interventions capable of inducing A in tumor cells is provided. These interventions are of potential therapeutic value. The balance of apoptotic and anti-apoptotic forces in virally infected normal and malignant cells is discussed. PMID- 11494025 TI - Comparative genomic hybridization analysis of chromosomal alterations in patients with long-standing ulcerative colitis. AB - Patients with ulcerative colitis (UC) are prone to develop colorectal cancer which is related to the duration and extent of the disease. One of the earliest events in tumor progression is the development of aneuploidy. Aneuploidy is correlated with the grade of dysplasia which serves as a common but not always reproducible marker for the prediction of UC associated formation of cancer. We analyzed 48 biopsy samples from 5 patients with long-standing ulcerative colitis by comparative genomic hybridization (CGH). The majority of these samples represented premalignant stages which are not well characterized at the molecular level as yet. We compared biopsy samples from different colon locations in regard to chromosomal alterations, dysplasia status and DNA index. Besides chromosomal changes occurring only in certain patients in restricted areas of the colon we also detected amplifications and deletions which were common in all persons throughout the colon. The stage of dysplasia seems to have no influence on the number and appearance of chromosomal changes. Amplifications in 2, 3, 6, 9, 11, 12 and 15 were found in almost all cases. In dysplastic samples chromosomal regions 3, 6 and 11 revealed gains of DNA. Deletions were detected within 8q, 15, 18q, 20p and 22q. The affected chromosomal regions may contain yet unknown oncogenes or tumor suppressor genes participating in UC associated carcinogenesis. The conspicuous regions found in the CGH experiments allow the selective and detailed characterization at a molecular level. PMID- 11494026 TI - Expression of the IL-6 family cytokines in human brain tumors. AB - In our RT-PCR screen for cytokine expression in human brain tumors we discovered increased levels of oncostatin M (OSM), ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF), all belonging to the interleukin-6 (IL-6) cytokine family, in most of the tumors. The expression of these cytokines in normal adult brain tissue was found to be very low or below detection limit. OSM expression was elevated in most of the tumors and immunohistochemistry analysis showed that the tumor cells contained OSM in their cytoplasm, suggesting they produce this factor. Overexpression of OSM has not previously been reported in primary human brain tumors. The IL-6 cytokine family acts through a common gp130 receptor subunit that activates the JAK/STAT signaling pathway and therefore they have been suggested to have overlapping effects. Tissue inhibitor of metalloproteinase-1 (TIMP-1), matrix metalloproteinase 1 (MMP-1) and MMP-3 and IL 6 have been reported to be regulated by OSM. IL-6 was low or absent in the tumors. TIMP-1, MMP-1 and MMP-3 was expressed in most tumors but with no strict correlation to OSM levels. PMID- 11494027 TI - Serum anti-p53 in relation to mutations across the entire translated p53 gene in colorectal carcinomas. AB - Previous reports have claimed that antibodies to mutated p53 protein indicate poor outcome in malignant disease. The mechanism behind this highly specific process is unclear, although it has been claimed that certain DNA alterations are prone to induction of immune response, since wild-type p53 is almost never immunogenic. The aim of the present analysis was to evaluate whether the presence of anti-p53 was statistically significantly related to any certain DNA alterations in the entirely expressed p53 gene in primary tumors of colorectal cancer. P53 serum antibodies were determined by an enzyme linked immunosorbant assay (ELISA). P53 antibodies were detected in serum of 24 of 88 patients (27%). Twenty-two of 24 (92%) sero-positive patients had mutations in their p53 gene while only 22 of 64 (34%) sero-negative patients had p53 mutations (p<0.01). Mutations were mainly missense with a trend to significantly higher frequency of deletions in sero-negative patients compared to sero-positive subjects (8/25, 32% and 2/22, 9% respectively, p<0.08). Mutations in sero-positive patients were mainly located in exon 5 and 7 and within conserved regions (17 of 22 mutations). In sero-negative patients missense mutations were usually located in exon 5, 7 and 8 being also most frequently located within conserved regions. Most of the p53 deletions in sero-negative patients were however located outside conserved regions (seven of eight deletions). There was no statistical difference between sero-positive and negative patients concerning the spectrum of mutations along the expressed gene. Our study demonstrates that p53 antibodies are usually related to p53 gene mutations but a mutational event is not sufficient to elicit self-immunization. Cellular protein binding to p53 or individual differences of major histocompatibility complex based presentation of p53 protein sequences by immune cells is therefore the most likely explanation between sero-negative and sero-positive patients. PMID- 11494028 TI - Expression of telomeric repeat binding factor 1 and 2 and TRF1-interacting nuclear protein 2 in human gastric carcinomas. AB - Telomeric repeat binding factor 1 (TRF1) and 2 (TRF2) may play key roles in the maintenance of telomere function. TRF1 negatively regulates telomere elongation, while TRF2 protects the chromosome ends by inhibiting end-to-end fusions. We examined the expression of TRF1 and TRF2 in 20 gastric carcinomas by reverse transcription polymerase chain reaction and then analyzed the relation with telomerase activity and other telomerase components such as human telomerase reverse transcriptase (TERT), human telomerase RNA component (hTR), human telomerase-associated protein (TEP1) and TRF1-interacting, ankyrin-related ADP ribose polymerase (tankyrase) as well as TRF1-interacting nuclear protein 2 (TIN2). Of 20 gastric carcinomas examined, 10 (50%) and 12 (60%) expressed TRF1 and TRF2 at higher levels than did non-neoplastic mucosa, respectively. No obvious correlation was observed between TRF1 expression and telomerase activity or expression of TERT, hTR and TEP1. Carcinomas with high TRF1 expression expressed significantly higher levels of tankyrase and TIN2 than did those with low TRF2 expression (p<0.05). The telomerase activities and the levels of TERT, hTR and TEP1 showed tendency to be lower in tumors expressing TRF1 at low levels, although it was not significant. On the other hand, carcinomas with short telomere length (shorter than 2 Kbp) expressed significantly stronger telomerase activities and higher TRF1 expression (p<0.05) and tended to express TRF2 and TIN2 at higher levels than those with long telomere length. The results suggest that gastric carcinomas with short telomeres need high levels of telomerase activity and large quantity of TRFs and TIN2, whereas those with long telomeres do not require high levels of telomerase activity and telomere associated proteins. PMID- 11494029 TI - Microsatellite alterations in human hepatocellular carcinoma infected with hepatitis B virus: associated with the elevation of serum alpha-fetoprotein. AB - Identification of the basic genetic changes in human hepatocellular carcinoma (HCC) is very important for the understanding of this cancer. In this study, genomic DNA from 29 pairs of HCC and corresponding non-tumour tissues infected with hepatitis B virus (HBV) was prepared. Five CA-repeated microsatellite markers, including D8S277, D3S1029, D5S409, D2S123, and TP53, were used to analyse microsatellite alterations and their subtypes in these patients by polymerase chain reaction (PCR) amplification and denatured polyacrylamide gel electrophoresis. Microsatellite alterations were found in 15 of the 29 HCC patients (51.72%), implying that microsatellites are unstable in genomic DNA of HBV-infected HCC. It was found that frequency of microsatellite alterations in these HCC patients was not associated with patients' age, sex, status of tumour differentiation, and tumour size. Frequency of microsatellite alterations in HCC patients with cirrhosis tended to be less than that in patients without cirrhosis, but Fisher's exact test, 2-tailed, showed that this difference was not significant. Significantly more microsatellite alterations in serum alpha fetoprotein (AFP)-positive cases were observed than those in serum AFP-negative ones, implying that the elevation of AFP in HBV-infected HCC may be associated with microsatellite stability. PMID- 11494030 TI - Elevated levels of cathepsin B in human glioblastoma cell lines. AB - Degradation of the extracellular matrix is a prerequisite for the invasive phenotype in glioma cells. Several proteases released by invading tumor cells seem to participate in the focal degradation of extracellular matrix proteins. Using enzymatic assays, Western blotting, and Northern blotting techniques, we investigated whether cathepsin B level was associated with malignant grade in seven human glioma cell lines. Cathepsin B activity and protein content levels were higher in glioblastoma cell lines than in anaplastic astrocytoma or low grade glioma cell lines. Cathepsin B transcripts were overexpressed in glioblastoma cell lines relative to their expression in anaplastic astrocytoma and low-grade glioma cell lines. Cathepsin B promoter activity and amount of SP-1 complexes were much higher in glioblastoma cell lines than in anaplastic astrocytoma or low-grade glioma cell lines. Finally, E-64, an inhibitor of cathepsin B, inhibited both cathepsin B enzymatic activity and the invasiveness of glioblastoma cell lines. These results strongly support a role for cathepsin B in glioblastoma cell lines and suggest that inhibition of cathepsin B activity may be proven useful in cancer therapy. PMID- 11494031 TI - The inhibitory effect of simvastatin on growth in malignant gliomas--with special reference to its local application with fibrin glue spray in vivo. AB - Simvastatin is one of the competitive inhibitors of HMG-CoA reductase. During clinical trials, it has shown the ability to lower serum cholesterol. We investigated the effect of simvastatin on the growth of malignant gliomas in vitro, semi-in vivo, and in vivo. An in-vitro MTT assay revealed that human malignant glioma cell lines: U-251MG, U-373MG, and U-87MG, and rat malignant glioma cell line C6 were well inhibited in growth in a dose-dependent fashion. An anchorage-independent growth assay showed that the number of colonies (more than 100 microM in size) of human (U-373MG) and rat malignant gliomas (C6) was markedly reduced in a dose-dependent fashion. A flow cytometry analysis revealed that simvastatin treatment led U-251MG cells to accumulate in sub G0-G1. Immunostaining by TUNEL method showed that most glioma cells treated by 10 microM simvastatin had nuclear immunostaining, suggesting apoptotic changes of the treated cells. The human umbilical vein endothelial cells and human lung fibroblasts were inhibited in growth by no more than 20% of controls even with a high dose (10 microM) of simvastatin. In the semi-in vivo model, using newborn rat brain slice cultures, the rhodamine-labeled glioma cells were abolished after 7 days of local simvastatin treatment with fibrin glue probably suggesting that simvastatin led the cells to apoptosis. In rat models using subcutaneously inoculated C6, the local application of simvastatin combined with fibrin glue (spray method) was quite effective in inhibiting the growth of the tumor. These data suggest that simvastatin may be a novel anti-glioma drug, and the local application of simvastatin combined with fibrin glue (by spray method) may be a crucial new clinical strategy against glioma growth. PMID- 11494032 TI - Up-regulation of WNT10A by tumor necrosis factor alpha and Helicobacter pylori in gastric cancer. AB - WNT signaling pathway is implicated in carcinogenesis and embryogenesis. We have previously cloned and characterized WNT10A and WNT6, which are clustered in human chromosome 2q35 region. In this study, we investigated expression of WNT10A and WNT6 in gastric cancer. The 3.0- and 2.4-kb WNT10A mRNAs were expressed in gastric cancer cell lines MKN7, MKN45 and MKN74. The 2.0-kb WNT6 mRNA was expressed in gastric cancer cell lines MKN28 and MKN74. WNT10A was up-regulated in 3 out of 6 cases of primary gastric cancer, while WNT6 was not up-regulated in primary gastric cancer. Effects of inflammatory cytokines and Helicobacter pylori (H. pylori) on expression of WNT10A and WNT6 were next investigated. Interferon gamma (IFNgamma) failed to induce up-regulation of WNT10A and WNT6. Tumor necrosis factor alpha (TNFalpha) induced up-regulation of WNT10A in MKN45 cells. Up-regulation of WNT10A reached maximum at 6 h after TNFalpha treatment. H. pylori also induced up-regulation of WNT10A in MKN45 cells. These results strongly suggest that up-regulation of WNT10A induced by TNFalpha and H. pylori might play key roles in human gastric cancer through activation of WNT--beta catenin--TCF signaling pathway. PMID- 11494033 TI - Survivin, bcl-2 and matrix metalloproteinase-2 enhance progression of clear cell- and serous-type ovarian carcinomas. AB - Clear cell adenocarcinomas (CA), unlike serous adenocarcinomas (SA) of the ovary, are often at stage I, are resistant to platinum-based drugs and have a poor prognosis. The causes of these differences are unclear. In this study, the differences in progression between CA and SA were examined in terms of apoptosis related and tumor invasion-related factors. The 16 cases of CA and the 16 cases of SA were reviewed. Excised tissues were classified into primary or metastatic loci, and the expressions of survivin, Bcl-2 and matrix metalloproteinase-2 (MMP 2) in each locus immunohistochemically assayed. Whether the expression of each protein was correlated to prognosis was investigated and additionally the invasion ability of cell strains established from CA and SA were examined using in vitro invasion assay. CA at stage I showed significantly higher survivin expression than SA (p<0.05). In CA, survivin tended to be expressed higher in primary locus than in metastatic locus (p=0.068), however, Bcl-2 was expressed relatively higher in the latter (p=0.087). SA did not have these tendencies. While MMP-2 was expressed significantly higher in SA than in CA (p<0.05), and more so in metastatic locus than in primary locus of SA (p<0.05). Invasion assay showed that the invasion of cells derived from SA was significantly inhibited by tissue inhibitors of metalloproteinase-2, an MMP inhibitor. The disease-free interval was significantly shorter when survivin expression was observed in the nucleus. These results suggest that the expression of apoptosis inhibiting factors and enhanced invasion ability affect progression of CA and SA, respectively. PMID- 11494034 TI - Fluorescence in situ hybridization to assess transitional changes of aneuploidy for chromosomes 7, 8, 10, 12, 16, X and Y in metastatic prostate cancer following anti-androgen therapy. AB - There have been few detailed studies conducted on the cell population in relation to cytogenetic changes between the pre- and post-treatment periods in patients with prostate cancer. We investigated numerical chromosome changes associated with anti-androgen therapy, using fluorescence in situ hybridization (FISH). FISH using chromosome-specific centromeric probes was used to assess transitional changes in the frequency of aneuploidy for chromosomes 7, 8, 10, 12, 16, X, and Y in prostate cancer during the pre- and post-treatment periods. Gains of chromosomes 7, 8 and 12 were notable in the pre-treatment samples (8 out of 9 cases in chromosome 7; 8 out of 9 cases in chromosome 8; 7 out of 9 cases in chromosome 12), while a notable reduction in the number of cells with extra copies of these chromosomes was observed in post-treatment specimens. Other chromosomes did not show noticeable change in their FISH signals at each phase of clinical treatment in all 9 cases. Changes in cell number with high ploidies of chromosome 7, 8 and 12 reflect the clinical effects of anti-androgen therapy at the early phase, which might explain the androgen dependency of metastatic prostate cancer cells. PMID- 11494035 TI - P53 and hMSH2 expression in basal cell carcinomas and malignant melanomas from photoexposed areas of head and neck region. AB - Ultraviolet (UV) radiation plays a pivotal role in skin damage and photocarcinogenesis. The basic mechanism of phototoxicity lies in DNA damage, and involves mutation of tumor suppressor genes, oncogenes and genes directly involved in the control of the stability of genome, such as the mismatch repair (MR) genes. The goal of this study was to evaluate the role of p53 and hMSH2 in the UV-related carcinogenetic process. An immunohistochemical study for p53 and hMSH2 was performed in a series of 43 basal cell carcinomas (BCC) and 60 melanomas (MM) from photoexposed areas of head and neck region, comparing the findings with follow-up. A deregulated p53 expression characterized less differentiated, more aggressive BCC (BCC2) but not the well-differentiated ones (BCC1). The hMSH2 protein was present, though expressed at varying levels, in 18 out of 21 BCC1 cases and in 4 out of 22 BCC2. In the remaining 3 cases of BCC1 and 18 cases of BCC2, a complete absence of hMSH2 expression was found, correlating directly with the presence of recurrence and/or death of the disease in case of melanoma (p<0.05). Overall, the expression of hMSH2 correlated inversely with the p53 overexpression (p<0.01). In MM, p53 was found overexpressed in 81.6% of the cases, and this correlated positively with the level of infiltration and with the presence of relapses (p<0.01) or metastasis (p<0.01) and inversely with the disease-free interval (p<0.05). These results are in agreement with the reported association between p53 deregulation and a more aggressive cancer phenotype. The evaluation of the expression of p53 and hMSH2 could improve the management of patients with BCC and MM, and could have a role also in the evaluation of the early cutaneous photo-inducted damage, contributing to the identification of presymptomatic patients predisposed to the development of UV-related new skin tumors, who could become candidates for chemoprevention trials. PMID- 11494036 TI - CD3+CD56+CD8+ cells demonstrating a suppressor T cell-like function in the peripheral blood of colon cancer patients. AB - We previously reported that HLA-unrestricted CTLs against MUC-1 were induced from colon cancer patients by stimulating peripheral blood lymphocytes (PBLs) with recombinant MUC-1 vaccinia virus (rVMUC-1). We have performed adoptive immunotherapy (AI) for two gastric and two colon cancer patients, using the rVMUC 1-stimulated T lymphocytes. A significant level of HLA-unrestricted cytotoxicity against MUC-1 was induced in the two colon cancer patients (pA and pB) during the first adoptive immunotherapy, but extremely reduced during the second AI. During the second stimulation phase, the rate of CD3+CD56+CD8+ cells were significantly increased and that of CD3+CD56-CD4+ cells were significantly decreased in the two colon cancer patients as compared to the first AI. CD3+CD56+CD8+ and CD3+CD56 CD4+ cells were isolated from the second AI of the colon cancer patient (pB) and designated as D856 and D4, respectively. The D4 cells demonstrated a high level of HLA-unrestricted CTL activity against MUC-1, but D856 cells did not. When D856 cells were mixed with D4 cells at a D856/D4 ratio of 1:3, 1:2, and 1:1 and used as effector cells, the HLA-unrestricted and MUC-1-specific CTL activity of D4 cells was suppressed in a D856/D4 ratio-dependent manner. Further, D856 cells were highly lytic for the D4 cells demonstrating HLA-unrestricted cytotoxicity against MUC-1. It is concluded that the reduction in HLA-unrestricted cytotoxicity against MUC-1 during the second AI is attributed to the D856 cells killing MUC-1-specific CTLs (D4). Thus, the CD3+CD56+ CD8+ cells seem likely to behave as a suppressor T cell. PMID- 11494037 TI - hMLH1 and hMSH2 expression in human hepatocellular carcinoma. AB - The role of microsatellite instability (MSI) in the pathogenesis of hepatocellular carcinoma (HCC) is incompletely defined. Although high-frequency MSI (MSI-H) is infrequently seen in HCC, some studies have suggested a role for MSI in HCC development. While MSI has been clearly defined for a subset of tumors, in particular colorectal, gastric and endometrial cancers, generally accepted criteria have not been developed for other tumors. Colorectal cancers (CRC) are classified as MSI-H if >30-40% of >5 microsatellite loci analyzed show instability. The MSI-H phenotype is associated with defective DNA mismatch repair (MMR) and is observed in the majority of tumors from patients with hereditary non polyposis colon cancer (HNPCC) and also in 15% of sporadic CRCs. Inactivating mutations of the hMLH1 or hMSH2 genes lead to defects in MMR in HNPCC. In sporadic CRCs, MMR is usually due to hypermethylation of the hMLH-1 promoter. The role of defective MMR in hepatocellular carcinogenesis is controversial. Immunohistochemistry for hMLH1 and hMSH2 reliably indicates hMLH1 or hMSH2 loss in MSI-H CRC tumors. To investigate the role of defective MMR in HCC carcinogenesis, we performed immunohistochemistry for hMLH1 and hMSH2 on 36 HCCs. BAT26, a microsatellite marker that reliably predicts MSI-H was also examined. All 36 of the tumors stained positively for both hMLH1 and hMSH2, strongly suggesting an absence of either inactivating mutations of hMLH1 and hMSH2 or promoter hypermethylation of hMLH1. None of the tumors showed MSI at the BAT26 locus. These findings suggest that defective MMR does not contribute significantly to hepatocellular carcinogenesis. PMID- 11494038 TI - Targeting of doxorubicin to ES-2 human ovarian cancers in nude mice by linking to an analog of luteinizing hormone-releasing hormone improves its effectiveness. AB - Receptors for luteinizing hormone-releasing hormone (LHRH), expressed by ovarian cancers, can be used for targeting chemotherapeutic compounds more selectively to these tumors. We investigated the effects of cytotoxic LHRH analog AN-152, consisting of doxorubicin (DOX)-14-O-hemiglutarate linked to the epsilon-amino group of [D-Lys6]LHRH, on the growth of LHRH receptor-positive ES-2 human ovarian cancer line xenografted into nude mice. A single injection of AN-152, at a dose of 345 nmol/20 g body weight, caused a 34.5% reduction (P<0.05) in tumor growth after 28 days, while its cytotoxic moiety DOX was inactive at the same dose. Since the overexpression of certain growth factors and/or their receptors, such as vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR) and HER-2/neu, as well as various oncogenes like c-fos and c-jun, is associated with unfavorable prognosis and contributes to progressive growth of ovarian carcinomas, their mRNA levels were analyzed by RT-PCR. Treatment with AN 152 significantly (P<0.05) reduced the expression of EGFR, VEGF, c-fos and c-jun, to 49%, 48%, 55% and 58% respectively, compared to controls. HER-2/neu mRNA expression was also decreased to non-detectable levels. Conversely, DOX decreased non-significantly the expression levels for EGFR by 32%, VEGF 35%, both c-fos and c-jun approximately 20% and HER-2/neu by only 15%. In conclusion, cytotoxic LHRH analog AN-152 could be considered for chemotherapy of ovarian cancers expressing LHRH receptors. PMID- 11494039 TI - Effects of tetramethylpiperidine (TMP)-substituted phenazines on membrane stability and P-glycoprotein function. AB - The lipophilicity and membrane-destabilizing activities of clofazimine and three tetramethyl-piperidine (TMP)-substituted phenazines were compared with the anti tumor and multiple drug resistance (MDR) neutralizing potential of these agents using a P-glycoprotein (P-gp)-expressing small cell lung cancer cell line (H69/LX4). Partition coefficients were measured as an index of lipophilicity, while membrane-destabilizing potential was measured using a conventional hemolytic assay. The membrane-destabilizing potential of the TMP-substituted phenazines was found to correlate positively with the degree of lipophilicity, as well as with MDR reversal activity. The presence of a TMP group, as well as chlorine atoms on the phenyl and anilino rings of these agents contributed to the enhancement of anti-tumor activity by potentiating membrane-destabilizing activity. TMP-substituted phenazines may be useful in the design of novel anti cancer and MDR reversal agents. PMID- 11494040 TI - Effect of thermosensitive liposomal doxorubicin with hyperthermia on primary tumor and lung metastases in hamster osteosarcoma. AB - We evaluated the effect of intravenous thermosensitive liposomal doxorubicin (TL DOX) together with local hyperthermia on primary tumors in highly metastatic hamster osteosarcoma. This combination resulted in higher DOX concentrations in plasma, primary tumors and lungs than standard DOX under the same conditions. Tumor growth and lung metastasis were also inhibited more by TL-DOX and hyperthermia than by hyperthermia alone, DOX with or without hyperthermia, and TL DOX without hyperthermia. In addition, gains in hamster body weight were not suppressed. These results suggest that the combination of TL-DOX and hyperthermia can control primary tumors and suppress lung metastasis in hamsters. PMID- 11494041 TI - A novel role of tissue factor pathway inhibitor-2 in apoptosis of malignant human gliomas. AB - Tissue factor pathway inhibitor-2 (TFPI-2) is a 32 kDa serine protease inhibitor found at high levels in extracellular matrix. Recombinant human TFPI-2 has recently been shown to be a strong inhibitor of trypsin, plasmin, plasma kallikrein, and factor XIa amidolytic activity. Earlier studies in our laboratory showed that the expression of TFPI-2 is lost during tumor progression in human gliomas. We stably transfected this protease inhibitor in multiform glioblastoma cell line (SNB-19) and in low-grade glioma cell line (Hs683) in sense and antisense orientation respectively. This confirmed that the upregulation/down regulation of TFPI-2 plays a significant role in the invasive behavior of human gliomas both in vitro and in vivo models. Collectively, these results suggested an idea to determine whether TFPI-2 is necessary for cell survival and inhibition of tumor formation in nude mice, due to apoptosis of intracerebrally injected SNB 19 cells. In the present study we determined p-ERK levels and found that they are decreased in TFPI-2 over-expressed clones (SNB-19) and increased in TFPI-2 down regulated clones (Hs683). We also checked the levels of BAX/BCl-2, caspases (for e.g., 9, 7, 3, 8), PARP, cytochrome-c and Apaf-1. Moreover, the increase of apoptosis in vitro is associated with increased and decreased expression of apoptotic protein BAX in sense clones (SNB-19) and antisense clones (Hs683) respectively, when compared to controls and vice versa with Bcl-2 the anti apoptotic protein. Caspases (9, 7 and 3), cytochrome-c, Apaf-1 and PARP levels are increased in SNB-19 and decreased in Hs683. Caspase 8 was not expressed in either cell line. Caspases 9 and 3 activity assay revealed higher activity in sense clones (SNB-19) but lesser in antisense clones (Hs683) compared to controls. This is the first report of TFPI-2 playing a novel role in cell survival in human gliomas. PMID- 11494042 TI - Molecular pathways involved in response to ionizing radiation of ID-8 mouse ovarian cancer cells expressing exogenous full-length Brca1 or truncated Brca1 mutant. AB - BRCA1 germline mutations have been linked to the development of hereditary breast and ovarian cancers. Recent studies suggest that BRCA1 may function in the regulation of basic cellular processes, including gene transcription, and sensing and/or repair of DNA damage. To further delineate the BRCA1 upstream and downstream steps involved in its role in the cellular response to ionizing radiation, we compared the effects of expression of an exogenous full-length Brca1 with those of a truncated Brca1 mutant in the ID-8 mouse ovarian cancer cell line after irradiation. We found that expression of both full-length and truncated Brca1 increased resistance to ionizing radiation. Expression of truncated, but not full-length, Brca1 then allowed us to identify new potential downstream targets of mutated BRCA1 like MAPK/ERK pathway members and also key genes involved in mutated BRCA1 signaling pathway response to ionizing radiation such as p53 and p21WAF1/CIP1. We therefore established an in vitro mouse model for studying the molecular effects of human BRCA1 germline mutations. PMID- 11494043 TI - Absence of mutation of the p73 gene in astrocytic neoplasms. AB - In subgroups of astrocytic neoplasms, including glioblastoma (GBM), mutations of the p53 tumour suppressor gene lead to loss of growth-suppressive properties. A p53-related gene termed p73 has recently been identified; its gene product shows structural and functional similarities to p53. After being mapped to chromosome region 1p36, p73 was proposed to act as a tumour suppressor gene, as this region is frequently deleted in a variety of human cancers, including astrocytic tumours. To determine whether p73 is involved in astrocytoma/GBM development, we analysed 10 pilocytic astrocytomas, 15 WHO grade II astrocytomas, 15 WHO grade III anaplastic astrocytomas, and 20 GBM for p73 gene alterations. In parallel, we used six polymorphic markers to determine the allelic status of region 1p36 in this tumour series. Although loss of heterozygosity was evidenced in 12 of 60 cases (20% of samples), PCR-SSCP and direct sequencing failed to detect any gene mutation in the entire coding region and intronic sequences of p73. Eight tumours displayed five distinct polymorphic nucleotide changes, also present in the corresponding normal DNA. These variations consisted of T-->C variation, with no change in Thr173; C-->T transition, with no change in His197; exon 9 simultaneous double change C-->T and T-->C , with no variations in Ala336 and His349, respectively, and C-->T change at exon 9/-24 position of intron 8. These results suggest that, in astrocytic gliomas, p73 may not play a major role as a tumour suppressor, but the relatively high incidence of LOH confirms the presence at 1p36 of an as yet unidentified gene of this category, with a key function in astrocytoma/GBM progression. PMID- 11494044 TI - DNA alkylation-induced phosphorylation of p53 and activation of kinases in colon cancer cells. AB - In response to DNA damage, p53 protein transiently stabilizes and accumulates in the nucleus, where it performs its role as a transcription factor. Phosphorylation of p53 increases its sequence-specific DNA-binding activity. In the present study, we have examined the effect of methylmethane sulfonate (MMS) to HCT-116 human colon cancer cells on the phosphorylation of p53. Results show that p53 protein becomes phosphorylated at serine 15 (Ser15) and Ser392 residues after treatment with MMS in a time-dependent manner. Increased levels of phospho p53(Ser15) and phospho-p53(Ser392) were maintained up to 50 h of the MMS treatment. We also examined the involvement of probable kinase(s), which could be responsible for MMS-induced phosphorylation of p53 at Ser15 and Ser392. In vitro phosphorylation assay, carried out with the immunoprecipates of MMS-treated cells, showed an increased phosphorylation of p53 by c-Jun kinase 1 (JNK1) at early time points (2.5 h). However, with cyclin-dependent kinase (Cdk2) and TFIIH complex associated kinase CAK, the phosphorylation of p53 was increased at later time points (25 h). The phosphorylation of p53 by Cdc2 and MAPK (p38) kinases remained unaffected in the MMS-treated versus untreated cells. The MMS-induced phosphorylation of p53 correlates with our previous findings of p53's ability for increased sequence-specific DNA-binding and transcriptional activity in the cells treated with DNA alkylating agents. PMID- 11494045 TI - FISH monitoring of 100 courses of human leukemias: the cytogenetic viewpoint. AB - Interphase fluorescence in situ hybridization (I-FISH) analyses were performed from 2 up to 13 times along the course of 100 human leukemias (36 chronic myeloid leukemias, 38 acute myeloblastic leukemias, 17 acute lymphoblastic leukemias, and 9 additional hematopoietic neoplasias) in order to control clonality, evolution, and disappearance of the basic cytogenetic changes. The relevance of these data to confirm clinical remission or to detect minimal residual disease and/or relapse was evaluated. Fifty-four patients were monitored following hematopoietic stem cell transplantation, and 46 cases after chemotherapy. Various chromosome or aberration specific DNA probes were applied for follow-up in the time frame of 1 month up to 13 years. From the cytogenetic point of view, the aim was to determine the power of resolution of the I-FISH technique in the detection of clinically significant changes in the course of disease and its usefulness in daily routine cyto-genetics as compared with classical cytogenetics. In addition, reliability standards of the various DNA probes should be established. In 75 patients with remissions during the entire period of I-FISH monitoring no conspicuous signal constitution was detected. Of 25 relapses or progresses of disease, which were clinically confirmed, 22 were reliably detected by I-FISH, in only 1 case I-FISH monitoring failed to detect the aberrant clone. In 2 patients conventional karyotype analysis confirmed the relapse by detecting complex chromosomal aberrations, but specific probes for I-FISH confirmation were not available. These data suggest that I-FISH analyses in the follow-up of leukemias is a simple and in most cases sufficiently sensitive and highly reliable way of monitoring the outcome of therapy. It may well serve to close the gap between conventional karyotyping and the highly sensitive molecular techniques. PMID- 11494046 TI - Expression of alternatively-spliced MDM2 transcripts in giant cell tumours of bone. AB - Giant cell tumours of bone (GCT) are characterized histologically by multinucleated bone resorbing giant cells in a background of ovoid spindle-shaped mesenchymal cells. Current evidence suggests that the latter comprise the tumour element of these lesions, although there are basic questions as to the factors that contribute to the tumourigenesis and progression of GCT. The deregulation of the p53/MDM2 pathway is an important pathogenetic event in many tumour types, prompting us to assess the expression of MDM2 by the stromal cells and giant cells of GCT. Northern blot analysis demonstrated that most of the GCT samples examined expressed increased levels of MDM2 when compared to normal human bone cells. However, Southern analysis failed to show any evidence of MDM2 gene amplification in the same samples, suggesting that increased levels of MDM2 mRNA were not a direct result of gene amplification, but rather due to altered transcriptional regulation of MDM2 gene. By RT-PCR analysis we found that 7/8 giant cell tumours expressed strongly a short alternatively spliced variant of MDM2, whereas other tumours of bone and normal human bone cells expressed predominantly full length MDM2. Sequence analysis confirmed this variant to be MDM2-b, a variant previously reported to confer a transformed phenotype. Cell fractionation of the GCTs has shown that the MDM2-b splice variant was expressed exclusively in the stromal population, whereas the full length MDM2 was expressed in the multinucleated giant cells of these lesions. Overexpression of a green fluorescent protein-tagged MDM2-b in human embryonic kidney cells (HEK-293), demonstrated predominantly nuclear localisation. Immunoprecipitation studies showed that MDM2-b is unable to physically associate with the p53 tumour suppressor protein. These results are consistent with the hypothesis that the stromal cells comprise the tumour element in giant cell tumours of bone and we speculate that expression of the MDM2-b splice variant contributes to their transformed phenotype in a p53 independent manner. PMID- 11494047 TI - Photodynamic-induced apoptosis of human nasopharyngeal carcinoma cells using Hypocrellins. AB - It has been reported that novel photosensitizers Hypocrellin A and B, lipid soluble perylquinone derivatives of the genus Hypericum have a strong photodynamic effect on tumors and viruses. The molecular mechanisms of tumor cell death induction by Hypocrellin A and B are poorly understood. In this study, we have examined the photodynamic effects of Hypocrellin A and B compounds in poorly differentiated (CNE2) and moderately differentiated (TW0-1) human nasopharyngeal carcinoma (NPC) cells. Using these cell lines we investigated the role of the apoptotic pathway in photosensitized Hypocrellin A and B-mediated cell death. Tumor cells photoactivated with Hypocrellin A and B showed cell size shrinkage and an increase in the sub-diploid DNA content. A loss of membrane phospholipid asymmetry associated with apoptosis was induced by both tumor cell lines as evidenced by the externalization of phosphatidylserine (PS). A dose-dependent increase in caspases-3 protease activity inhibitable by the tetrapeptide inhibitor DEVD-CHO was also observed in both cell lines. Western blot analysis of poly (ADP-ribose) polymerase, a caspase substrate, showed the classical cleavage pattern (116 to 85 kDa) associated with apoptosis in Hypocrellin A and B-treated cell lysates. In addition, caspase inhibition blocked the externalization of membrane PS, indicating that the loss of membrane phospholipid asymmetry is a downstream event of caspases activation. These results demonstrate that tumor cell death induced by Hypocrellin A and B is mediated by caspase proteases. In conclusion, this study identifies both Hypocrellins (A and B) as potent and promising photosensitizers for the treatment of NPC. PMID- 11494048 TI - The diverse role of chemokines in tumor progression: prospects for intervention (Review). AB - Chemokines, proteins chemotactic for leukocytes and non-leukocytes, have been intensively studied for their role in tumor growth and metastasis. Recent work has shown that particular chemokines may have multiple effects on tumors including promoting growth, angiogenesis, metastasis, and suppression of the immune response to cancer, while other chemokines inhibit tumor mediated angiogenesis and promote anti-tumor immune responses. Increasing biological evidence supports the hypothesis that tumor-generated chemokines provide more than simply angiogenic signals. Tumor-derived chemokines may potentially act as inhibitors of anti-tumor immune responses as well as autocrine growth factors for the tumor. The complexity and redundancy of tumor chemokine expression suggests that a single chemokine target for tumor therapy may not be appropriate. Indeed, multiple target therapy including blockade of tumor enhancing chemokines while delivering or inducing the secretion of anti-tumor chemokines is the approach that currently holds the most promise. The role of chemokines in tumor biology as well as various means of blocking chemokines in cancer models in order to develop successful therapeutic strategies will be discussed. PMID- 11494049 TI - Galectin-8: a complex sub-family of galectins (Review). AB - Galectins are animal lectins, that can specifically bind beta-galactosides. Twelve galectins have been described in vertebrates, belonging to three different groups: prototype, tandem-repeat and chimeric. These proteins seem to be involved in cellular interactions and neoplastic transformations. We present an overview of a particular galectin member: galectin-8. This galectin, which has been intensively studied over the last six years, presents a particular type of gene regulation. It is widely expressed in tumoral tissues and seems to be involved in integrin-like cell interactions. Studies show that the LGALS8 gene encodes for almost seven mRNAs by alternative splicing pathways and various polyadenylation sites. These mRNAs could encode for six isoforms of galectin-8, of which three belong to the tandem-repeat galectin group (with two carbohydrate binding domains) and the three others to the prototype group (one carbohydrate binding domain). All these isoforms seem to be differentially expressed in various tumoral cells. This untypical galectin-8 subfamily seems to have a complex expression regulation, that could be involved in cancer phenomena. PMID- 11494050 TI - TGF-beta-induced p38 activation is mediated by Rac1-regulated generation of reactive oxygen species in cultured human keratinocytes. AB - p38 has been shown to be involved in TGF-beta-induced gene expression, but the upstream of the signaling pathway leading to the activation of p38 is left undefined. We investigated the pathway in cultured human keratinocytes (HaCat cells). Western blot analysis revealed that TGF-beta induced the activation of p38 within 1 h post TGF-beta treatment. H2O2 also strongly induced p38 activation in a time dependent manner. We also observed that TGF-beta-induced p38 activation was inhibited by PDTC, pyrrolidinedithiocarbamate, a known antioxidant, and DPI, diphenylene iodonium chloride, one of the known NADPH oxidase inhibitors. In contrast, TGF-beta-induced Smad2 phosphorylation was not affected. To test whether reactive oxygen species (ROS) is involved in TGF-beta-induced p38 activation, we examined the generation of ROS and activation of NADPH oxidase. FACS analysis showed that TGF-beta induced generation of ROS in time-dependent manner. DPI, an inhibitor of NADPH oxidase, inhibited TGF-beta-induced ROS production. Lucigenin-based NADPH oxidase assay indicated that TGF-beta-induced NADPH oxidase activity started as early as 5 min following treatment and peaked at about 15 min with induction of about 2-folds. The activity remained elevated up to 1 h. Immunofluorescence microscopy study showed that Rac1, one of the subunits of NADPH oxidase, translocated from cytoplasm to the membrane within 5 min. Pretreatment with DPI dramatically reduced TGF-beta-induced NADPH oxidase activity. Collectively, our data suggest that TGF-beta-induced p38 activation is mediated by Rac1-regulated generation of reactive oxygen species in cultured human keratinocytes. PMID- 11494051 TI - Mouse erythroblast formation is inhibited by anemia-inducing substance from the plasma of a patient with a malignant neoplasm. AB - An anemia-inducing substance (AIS) was found as a protein, with a molecular weight of 50,000 on SDS electrophoresis gel, that decreased the osmotic resistance of erythrocytes. In this study, the plasma fraction containing AIS is shown to inhibit mouse erythroblast formation in vitro. The addition of the plasma fraction from a patient with an advanced malignant neoplasm to a liquid culture of mouse bone marrow cells with erythropoietin results in low numbers of erythroblast formation and low cellular yields of alpha- and beta-globin mRNAs. These inhibitions were not observed after immunoadsorption of the plasma fraction with an antiserum against AIS. PMID- 11494052 TI - Calcitonin gene-related peptide (CGRP), acting via CGRP type 1 receptors, inhibits potassium-stimulated aldosterone secretion and enhances basal catecholamine secretion from rat adrenal gland. AB - Calcitonin gene-related peptide (CGRP) is a potent hypotensive peptide, that acts via two main subtypes of receptors, named CGRP1 and CGRP2. CGRP belongs to a regulatory-peptide family, that includes adrenomedullin (ADM) whose aldosterone antisecretagogue and catecholamine secretagogue actions are well demonstrated. Quantitative autoradiography showed the presence of [125I]CGRP binding sites in both rat adrenal zona glomerulosa (ZG) and medulla. Binding was displaced by the CGRP1-receptor antagonist CGRP(8-37), but not by the CGRP2-receptor agonist [cys(Et)2,7]-alphaCGRP (CGRP2-A). CGRP concentration-dependently inhibited 10 mM stimulated (but not basal) aldosterone secretion from dispersed rat ZG cells, and enhanced basal catecholamine secretion from rat adrenomedullary fragments. The responses to the maximal effective concentration of CGRP (10-8 M) were blocked by 10-7 M CGRP(8-37). CGRP2-A (10-7 M) neither altered aldosterone response to 10 mM K+ nor enhanced basal catecholamine secretion. The conclusion is drawn that CGRP, like ADM, inhibits agonist-stimulated aldosterone secretion and stimulates basal catecholamine release in the rat, exclusively acting via CGRP1 receptors. PMID- 11494053 TI - Angiogenic activity of leptin in the chick embryo chorioallantoic membrane is in part mediated by endogenous fibroblast growth factor-2. AB - Recently, it has been demonstrated that leptin, the product of the ob gene, playing a key role in the regulation of body weight, is angiogenic in vitro and in vivo. In this study we investigated the angiogenic potential of human leptin in vivo by using the chick embryo chorioallantoic membrane (CAM) assay, with the aim to establish whether this angiogenic activity is partly dependent on endogenous fibroblast growth factor-2 (FGF-2), which is normally expressed during CAM development. Results showed that leptin is able to stimulate angiogenesis and that the angiogenic response is similar to that obtained with FGF-2. The stimulating property of leptin is specific, as the application of anti-leptin antibodies onto the CAM significantly inhibits the angiogenic response. Moreover, this angiogenic activity is in part due to the activation of endogenous FGF-2. The application of anti-FGF-2 antibodies reduces the angiogenic response to leptin by 40%. Our study confirms that leptin is angiogenic in vivo and suggests that, at least in the chick CAM, its activity is in part mediated by the activation of endogenous FGF-2. PMID- 11494054 TI - Synergistic insulinotropic effects of succinic acid dimethyl ester and exendin-4 in anaesthetized rats. AB - It was recently proposed that suitable succinic acid esters could be used to potentiate the insulinotropic action of glucagon-like peptide 1 (GLP-1) in the treatment of type-2 diabetes mellitus. In such a perspective, the present study aimed mainly at investigating whether exendin-4 (Ex-4), a peptide structurally related to GLP-1(7-36)amide, and succinic acid dimethyl ester (SAD) also act synergistically upon insulin secretion in anaesthetized rats. Despite a higher plasma insulin concentration in SAD-infused rats (5.5+/-1.1 ng/ml) than in saline infused animals (1.9+/-0.7 ng/ml), the intravenous injection of Ex-4 augmented to a greater extent the plasma concentration of insulin in the former rats (+7.4+/ 2.5 ng/ml) than in the latter animals (+2.8+/-0.6 ng/ml). These findings document that the insulinotropic actions of Ex-4 and GLP-1 display comparable nutrient dependency, being both potentiated by a non-glucidic nutrient secretagogue such as SAD. PMID- 11494055 TI - Cloning and expression of uridine/cytidine kinase cDNA from human fibrosarcoma cells. AB - Uridine/cytidine kinase which converts uridine and cytidine to their corresponding monophosphates is a rate-limiting enzyme involved in the salvage pathway of pyrimidine synthesis. We isolated cDNA encoding the enzyme from human fibrosarcoma cells, then determined its nucleotide sequence by the 5'-RACE method followed by confirmation employing the human genome DNA library. The isolated uridine/cytidine kinase cDNA (UCK cDNA) consisted of 786 nucleotides encoding 261 amino acids and was found to have approximately 70% homology with mouse UCK cDNA. Northern blot analysis of human leukemia RNAs with labeled UCK gene showed a single band at 1.6 kb to be UCK mRNA, and southern blot analysis of the UCK cDNA after digestion with BamHI, SacI and XbaI enzymes showed four band signals, suggesting the UCK gene to have at least 4 exons. A truncated form of UCK cDNA was expressed as the His-tag conjugated protein in Escherichia coli. The expressed and purified protein specifically converted uridine and cytidine to their corresponding monophosphates and also phosphorylated antitumor nucleosides such as 5-fluorouridine, cyclopentenyl-cytosine and 3'-C-ethynylcytidine. The present results suggest that our cloned human UCK cDNA encodes the correct amino acid sequence for UCK protein, showing high intracellular phosphorylation activity forward natural and synthetic pyrimidine nucleosides. PMID- 11494056 TI - Expression of apoptosis regulatory proteins in the skeletal muscle of tumor bearing rabbits compared with diet-restricted rabbits. AB - The mechanism of body weight loss in the tumor-bearing state is still unclear. In this study, we investigated expressions of apoptosis regulatory proteins in the skeletal muscle of tumor-bearing and diet-restricted rabbits, and tried to evaluate the differences between the two groups. The apoptotic index (AI) in the tumor-bearing group was 28.1+/-2.84 on day 10. By day 20, many more apoptotic cells were found (AI: 40.5+/-3.20), but then after day 20 their numbers gradually decreased (AI: 9.67+/-2.22 on day 30 and 0.93+/-0.96 on day 40). By contrast, no apoptotic cells were detected in the diet-restricted group at any of the times examined. Bcl-2 immunoreactivity was either not detected at all or only weakly observed in both groups. By contrast, Bax expression increased gradually after implantation in the tumor-bearing group. Bax expression in skeletal muscle cell was graded (moderate) 10 days after tumor implantation, and (high) by day 20, in 2 of the 5 tumor-bearing rabbits. After day 20, however, Bax immunoreactivity decreased continuously in the tumor-bearing group. By contrast, hardly any Bax immuno-positive cells were detected in the diet-restricted group. These results suggest that loss of body weight in the tumor-bearing group is different from that in the diet-restricted group, and is related to apoptosis of skeletal muscles. PMID- 11494057 TI - Enzymic activities in two populations of purified rat islet beta-cells. AB - In terms of glucose sensing by pancreatic islet beta-cells, emphasis is currently placed on both the role of glucokinase, with negligible activity of low-Km hexokinase(s), and the prevalence of the oxidative over non-oxidative modality of glycolysis, a situation tentatively attributed, in part at least, to a low activity of lactate dehydrogenase. Conflicting information is available, however, on the activity of both low-Km hexokinase(s) and lactate dehydrogenase in purified beta-cell homogenates. This issue was reinvestigated, therefore, in two populations of purified rat islet beta-cells selected on the basis of their low (betaL) or high (betaH) content in reduced pyridine nucleotides. The size and protein content of betaH cells represented about twice that of betaL cells. Such was also the case for low-Km hexokinase(s), lactate dehydrogenase, mitochondrial FAD-linked glycerophosphate dehydrogenase, glutamate dehydrogenase and glutamate alanine and glutamate-aspartate transaminases. Whether in betaH or betaL cells, the activity of low-Km hexokinase(s) was at least as high as or higher than that of glucokinase. In both betaH and betaL, the activity of lactate dehydrogenase exceeded that required to catalyze the full reduction of glucose-derived pyruvate to L-lactate, as estimated from the rate of D-glucose phosphorylation under physiological conditions. These findings thus argue against a low expression of either low-Km hexokinase(s) or lactate dehydrogenase as major determinants of the glucose-sensing device in beta-cells. PMID- 11494058 TI - A new trend of breast cancer research in the genome era (Review). AB - The decipherment of the human genome, as accomplished recently in USA and in Europe, now enables us to search for the cause of a disease at the levels of the whole spectrum of gene expressions (mRNAs) of the human genome. A set of investigations came to the world in AD 2000 to show that: a) A total of 50 genes of the estrogen receptor positive (ER+) human breast cancer cell line MCF-7 in their gene expressions were found to undergo stimulation from a physiological concentration of 17beta-estradiol. b) Comparison of estrogen-responsiveness among a number of estrogen-responsive genes, among cancer cell lines of both the breast and endometrium with and without active ER, and among cell lines of normal tissues revealed that association between the presence of active ER and estrogen responsive genes is quantitative rather than qualitative including some exceptions, and that none of the estrogen-responsive genes tested was classified as of breast cancer specific. For this review, we collected information from our and other laboratories to investigate problems that remain to be disputed. Five items of discussion are given as follows: a) The dose of a steroid used for the production of an experimental tumor was fixed not to a physiological concentration but to a pharmacological concentration. In the case of estradiol, the latter was higher than the former by over 3 orders. The mitotic activity of MCF-7 underwent stimulation from the former but distinct suppression from the latter. b) A massive dose of a single steroid, when given at a good time of the host age, could produce a tumor of any kind. The timing of treatment rather than the nature of a steroid was found critical. c) Experience with the morphological development of Drosophila suggests the possibility that deficiency rather than amplification of gene expression in the infant age of Drosophila is responsible for the induction of morphological changes in an adult fly. Likewise, deficiencies of some escort steroids rather than overflow of estradiol may have more chance of occurrence in the genesis of spontaneous breast cancer, as suggested by many researchers including us. The plasma concentration of estradiol was found to be normal in patients with cancers of both the breast and endometrium. Future studies of breast cancer as well as other cancers should be directed to the multisteroidal carcinogenesis hypothesis rather than the monosteroidal carcinogenesis hypothesis. d) The necessity of recruiting an appropriate case-control data set and the difficulty of data interpretation were emphasized in the search for good biomarkers of breast cancer. e) Case-control studies of tamoxifen use was found useful for the prevention and clinical control of breast cancer of non-hereditary type but not for the breast cancer of hereditary type. Both decreased risk of breast cancer and increased risk of endometrial cancer were detected in the same population of tamoxifen use. The observed dualism of both human breast cancer and tamoxifen action can be taken as evidence to support the multi-steroidal carcinogenesis hypothesis rather than the mono-steroidal carcinogenesis hypothesis. PMID- 11494059 TI - Morphine reduces herpes simplex virus-1 pathogenesis in the murine flank. AB - Here we investigate the effect of morphine on herpes simplex virus-1 (HSV-1) pathogenesis using a murine flank scarification model. Murine flank scarification with HSV-1 results in primary lesions at the site of inoculation within three days and lesions at secondary sites within four days. The lesions are scored based on lesion size. Applying 0.1 mM morphine to the skin one-day post inoculation tested the effect of morphine on the formation of the herpes lesion. On days three through five, mice treated with morphine developed lesions with scores half of that observed in untreated animals, however, skin viral titers on these days were equivalent. Further, 1.0 microM morphine did not effect the replication rate of HSV-1 in Vero cells. Taken together, these data suggest the morphine reduced HSV-1 pathogenesis by modifying the host response to HSV-1 infection and not by reducing viral replication rates. PMID- 11494060 TI - Role of the third intracellular loop and of the cytoplasmic tail in the mitogenic signaling of the protease-activated receptor 1. AB - The activation of the protease-activated receptor 1 (PAR-1) by thrombin has been shown to induce an activation of the MAP kinase cascade and to stimulate cell proliferation. To examine the mechanisms of signal transduction by PAR-1, we constructed several PAR-1 mutants which were stably expressed in CHO cells. When compared to wild-type PAR-1, mutation of Ser306-->Ala (S306A) in the third intracellular loop of PAR-1 inhibited MAP kinase activation and cell proliferation stimulated by thrombin. The thrombin activation of MAP kinase was inhibited by pertussis toxin, suggesting a role for a Gi-like protein. As shown by calcium signaling and inosotol trisphosphate generation, the Ser306-mutated PAR-1 induced a strong activation of phospholipase C after thrombin addition. Deletion of the cytoplasmic tail of PAR-1 also inhibited thrombin-induced DNA synthesis but the MAP kinase pathway was activated as with wild-type PAR-1. In contrast, the deletion of the C-tail of PAR-1 prevented almost completely the activation of the phospholipase C pathway. Taken together these results suggest that the C-tail of PAR-1 is a critical site for PAR-1 coupling to phospholipase C activation, while the third intracellular loop of PAR-1 is implicated in PAR-1 coupling to Gi and MAP kinase activation. In addition, these results also show that MAP kinase activation is necessary but not sufficient for thrombin to induce cell proliferation. PMID- 11494061 TI - Zaltoprofen inhibits concanavalin A-induced decrease of body weight in mice. AB - Treatment of mice with concanavalin A (Con A) (12.5 mg/kg, i.v.) decreased the body weight at 24 h. This Con A-induced body weight decrease was accompanied by reduction in food and water intake. Zaltoprofen is a non-steroidal anti inflammatory drug. The administration of Zaltoprofen (10 mg/kg) at 8 h after Con A treatment was found to inhibit the Con A-induced reduction in body weight. The food intake in mice treated with Con A plus Zaltoprofen (10 mg/kg) was four times greater than that in mice treated with only Con A. The present results showed inhibition of the Con A-induced body weight loss by Zaltoprofen and suggested its possible effectiveness for the treatment of "sickness behavior". PMID- 11494062 TI - Metachromasia of the endocervical epithelium in women treated with gonadotropin releasing hormone agonist. AB - Endocervical epithelium from women treated with gonadotropin releasing hormone agonist (GnRH agonist) were compared with those from untreated women, using metachromatic stain with toluidine blue and immunohistochemistry, for estrogen and progesterone receptors. Smaller endocervical epithelium with lower intraepithelial mucus and more prominent nuclear distribution of estrogen receptors were seen in a woman who was treated for twelve months with GnRH agonists, as compared with those from an untreated premenopausal woman. PMID- 11494063 TI - Organizational commitment to integrated and collaborative management: matching strategies to constraints. AB - Integrated and collaborative approaches to environmental management are being advocated as a more appropriate and effective approach to decision-making. It is based on collaboration among a range of individuals and organizations that have a stake, role, or responsibility in management outcomes. However, researchers have found that implementation of this approach has encountered difficulties. This paper focuses on the role of organizations and their commitment to implementation. Based on case study and survey research in the United States and Australia, the author examines organizational constraints and the range of strategies used to secure commitment. It is argued that participants must more explicitly address the commitment issue and design implementation strategies that respond to organizational constraints. PMID- 11494064 TI - Conceptual design of monitoring and evaluation plans for fish and wildlife in the Columbia River ecosystem. AB - A logical sequence of seven steps is proposed as a generic template to design plans for monitoring and evaluating fish and wildlife in the Columbia River ecosystem. Management programs for these resources fail to include coordinated monitoring and evaluation plans. This short-coming is indicative of pervasive management conflicts detected from regional to local geographic scales. In the absence of a cohesive ecological management framework, monitoring and evaluation activities proceed without a clear understanding of what uncertainty they are intended to address, nor is there a clear description of the process to utilize the information gained. As a result, the accountability for the investment of public funds for fish and wildlife restoration is poor, information collected from the environment is not included in decision-making, and the ability to gain knowledge while taking management actions is compromised. The sequence of steps discussed here does not identify or describe distinct monitoring activities or methodologies at any particular location or listed under any specific monitoring plan. Instead, it concentrates on the generic elements necessary for the design and implementation of coordinated fish and wildlife monitoring plans. It is proposed that at least four major issues demand considerable attention in order to improve regional monitoring and evaluation capabilities: The first is adoption of an ecological framework for the management of fish and wildlife at relevant geographic scales within the ecosystem. Such a framework must include an explicit identification of goals, objectives, and actions to steer coordinated decisions across the boundaries of technical disciplines, management jurisdictions, and institutional responsibilities. The second is that the identification of these management goals for the geographic location of interest must precede the design of monitoring and evaluation plans from the top down. Third, the evaluation component must be considered early on in the planning process, so that it blends smoothly with monitoring at the time of implementation. Fourth, decision-makers and scientists engaged in the planning of fish and wildlife monitoring and evaluation efforts in the region must have a close collaborative relationship. Monitoring and evaluation plans designed under these premises may enhance our collective observational capabilities, promote cost-effectiveness and adequate evaluation, and provide a useful tool to adjust our management practices to the challenges of complex ecosystems. PMID- 11494065 TI - Jamaica's disappearing forests: physical and human aspects. AB - Jamaica is a small island that is losing its forest cover at a rapid rate. Due to the dependency of its largely poor population on the many services and functions its forests provide, this loss threatens to have substantial socioeconomic and ecological consequences for the country. Despite these basic facts, the problem of Jamaican deforestation has received very little attention from the scientific community. This article presents results of an island-wide, satellite-based study of forest change for Jamaica for the period 1987-1992, which was supplemented by a field trip to the island in 1999 to assess the overall accuracy of the estimate. Landsat MSS images, which are available only up until 1992, have proved to be an invaluable and cost-effective resource for mapping forest change in the tropics, particularly in large areas. A supervised classification indicates that Jamaica experienced an average annual deforestation rate of 3.9% for this period, a figure higher than existing estimates based on partial ground surveys but lower than the FAO's 1990 Tropical Forest Assessment of 5.3% for 1981-1990. Deforestation estimates for Jamaica's 14 parishes are also presented, based on the integration of satellite-derived forest classification maps with a parish administrative boundaries map of the island in a GIS. A correlation analysis between parish deforestation estimates and socioeconomic and land use/quality indicators derived from official sources suggests that deforestation is occurring most rapidly in highly populated areas possessing large numbers of small farmers who live and work under resource-poor conditions. By providing a sense of the magnitude of and main forest loss hotspots, it is hoped that these national and subnational level forest estimates will draw scientific attention to the problem of deforestation on the island. In addition, the socioeconomic analysis may provide policy-makers and planners with some sense of the relative contribution of underlying driving process in this deforestation as a first step toward the creation of effective social programs to combat the problem. PMID- 11494066 TI - Is risk associated with drinking water in Australia of significant concern to justify mandatory regulation? AB - Presently in Australia there are no mandatory drinking water standards. Here we argue that the risk associated with drinking water in Australia is of a dimension discernible to warrant mandatory regulations. The catchments that supply the major metropolitan areas of Sydney and Adelaide, and the groundwater for the city of Perth have been seriously compromised by the encroachment of development and activities. Melbourne in the past has generally relied on a closed catchment reservoir system; however, population growth in the near future will sequester the full online operation of additional reservoirs, which have multiple land use catchments. In addition to the current landscape circumstances, the management of a water system in itself proposes significant issues of risk. Two critical assumptions that are unique to a mass medium substance like water and dramatically alter the appraisal of risk are: (1) very large numbers of people are potentially exposed, and (2) small changes in contaminant levels may have adverse population outcomes. It is also known that water reticulation systems frequently suffer from contamination problems caused solely by the distribution system, and optimal management of these facilities would best be served by statutory protected transparency and dedicated water quality programs. In 1979, an Australian parliamentary committee stated that an "uncontaminated water supply is" a "basic requirement for the obtainment of good health"; however, recent surveys of Australian water systems show many are not meeting basic water quality criteria, and many communities are not receiving regular monitoring or testing as required by government authorized Australian drinking water guidelines. Exacerbating this situation is the lack of reporting and statutory endorsed standardized procedures to ensure information is properly and promptly recorded and that data are centralized for maximum benefit. The evaluation of risk associated with drinking water in Australia is often hampered by inadequate or incomplete data. Lastly, regional and rural water supplies face a vast array of contemporary problems and experiences that include widespread usage of pesticides and agricultural chemicals. In recent years, the Darling River has experienced the worst algal bloom known to man, and this river system not only supplies a number of regional and rural towns with water, but eventually connects with the River Murray, which supplies the State of South Australia with approximately 50% of its water requirements. PMID- 11494067 TI - Surface water pollution in three urban territories of Nepal, India, and Bangladesh. AB - In South Asian countries such as Nepal, India, and Bangladesh, pollution of rivers is more severe and critical near urban stretches due to huge amounts of pollution load discharged by urban activities. The Bagmati River in the Kathmandu valley, the Yamuna River at Delhi, and peripheral rivers (mainly Buriganga River) of Dhaka suffer from severe pollution these days. The observed dry season average of biochemical oxygen demand (BOD) in all these rivers is in the range of 20-30 mg/liter and total coliform are as high as 104-105 MPN/100 ml. Per capita pollution load discharge of urban areas has been estimated to be about 31, 19, and 25 g BOD/capita/day in Bagmati, Yamuna, and the rivers of Dhaka, respectively. Regression analysis reveals pollution loads steadily increasing nearly in step with the trend in urbanization. The dissolved oxygen (DO) level of the Bagmati and Buriganga rivers is declining at an average annual rate of nearly 0.3 mg/liter/year. Unplanned urbanization and industrialization occurring in these cities may be largely responsible for this grave situation. Inadequate sewerage, on-site sanitation, and wastewater treatment facilities in one hand, and lack of effective pollution control measures and their strict enforcement on the other are the major causes of rampant discharge of pollutants in the aquatic systems. PMID- 11494068 TI - Environmental indices in irrigation management. AB - Irrigation management calls for objective criteria capable of representing the economy, reliability, and productivity of irrigation systems. These criteria must be compatible with long-term sustainability and conservation goals. The criteria representing the above goals are the economic effect of management on yield reduction, economic effect, and reliability referring to plant growth and operation of the network. In this study environmental indices are introduced to express the above criteria in quantitative terms. The inclusion of these indices at the farm and network level create a multicriteria framework for decision making based on composite programming. An experimental study was conducted during the irrigation periods of 1989 and 1990 in Chania, Greece, concerning water delivered to 40 experimental plots, soil moisture content at the rootzone, and irrigation system operational failures. The data collected in real time were used for the calculation of the corresponding environmental indices. The variation in time and space is high and resulted in up to 62% of yield loss and low system performance (up to 7% of system temporal reliability). The study indicated that environmental indices could be incorporated to select alternatives and also to develop policies on water delivery. The final decision involves a trade-off analysis between cost of application and desired system performance. Measures of both primary objectives can be obtained using environmental indices that represent system operation aggregation at its basic levels (on farm and network). PMID- 11494069 TI - Public and occupational risks of the Nevada (USA) Test Site. AB - The Nevada Test Site (NTS), north of Las Vegas, was the scene of hundreds of nuclear weapons tests over four decades, both above- and below ground. There is considerable interest, both in neighboring communities and elsewhere, in the risks it poses. Overall, the greatest risks are nonradioactive in origin, with occupational risks to employees and accident risks in transporting low-level nuclear wastes to the NTS from other Department of Energy (DOE) sites ranking highest. For radiation risks, that to workers handling radioactive materials is much higher than that to the surrounding population, either present or future. Overall, annual risks are small, with all fatalities approximately 0.008% of total Nevada deaths. At the NTS, the government spends about 5000 times more on radiation as opposed to nonradiation deaths. This suggests that at least some resources may be misallocated towards cleanup of public risks and that the occupational risk of cleanup may be much higher than the public risk. Thus risk may be multiplied by well-meaning programs. PMID- 11494070 TI - Selecting suitable sites for animal waste application using a raster GIS. AB - Rapid growth of intensive animal industries in southeast Queensland, Australia, has led to large volumes of animal waste production, which possess serious environmental problems in the Murray Darling Basin (MDB). This study presents a method of selecting sites for the safe application of animal waste as fertiliser to agricultural land. A site suitability map for the Westbrook subcatchment within the MDB was created using a geographic information system (GIS)-based weighted linear combination (WLC) model. The factors affecting the suitability of a site for animal waste application were selected, and digital data sets derived from up to 1:50,000 scale maps were acquired. After initial preprocessing, digital data sets were clipped to the size of the delineated subcatchment boundary producing input factors. These input factors were weighted using the analytical hierarchy process (AHP) that employed an objectives-oriented comparison (OOC) technique to formulate the pairwise comparison matrix. The OOC technique, which is capable of deriving factor weight independently, formulated the weight derivation process by making it more logical and systematic. The factor attributes were classified into multiple classes and weighted using the AHP. The effects of the number of input factors and factor weighting on the areal extent and the degree of site suitability were examined. Due to the presence of large nonagricultural and residential areas in the subcatchment, only 16% of the area was found suitable for animal waste application. The areal extent resulting from this site suitability assessment was found to be dependent on the areal constraints imposed on each input factor, while the degree of suitability was principally a function of the weight distribution between the factors. PMID- 11494071 TI - Application of life-cycle assessment to type III environmental declarations. AB - The objective of this research is to develop a desirable type III environmental declaration format for the industrial consumer. A total of five different scenarios were chosen for the presentation of life-cycle assessment results in a format that meets the requirements of a type III environmental declaration. Life cycle assessment of an electronic component was performed for this purpose. Environmentally significant impact categories and input/output items of a product system derived from the life-cycle assessment are presented in this format. Presenting these information concisely in a standardized format would accelerate the communication of environmental aspects of materials and components in a product supply chain. Thus the type III environmental declaration can be a useful instrument for the manufacturing of an ecoproduct. PMID- 11494072 TI - Value to wildlife of urban-agricultural parks: a case study from Rome urban area. AB - Urban-agricultural parks could have some advantages to wildlife because of less intensive agricultural procedures, absence of hunting pressure, and reduced human disturbance. In this study, the breeding and wintering bird communities and the small mammal community in an urban-agricultural park of Rome were compared to those of a close urban park and a close agricultural area just outside the city. The aim was to assess the best destination and management of wildlife in natural areas at the urban-rural interface. Richness and diversity of bird communities were higher in the urban-agricultural park. Due to habitat features and probably human disturbances, but not to urbanization, predation, and competition factors, the urban park drastically reduces the abundance of decreasing open-land bird species. Abundance of these species was not significantly different in the urban agricultural park and in the agricultural area. In the urban-agricultural and urban park, bird and mammal pest species were more abundant than they were in the agricultural area. Regarding decreasing abundance of small mammal species, no significant difference among the study areas was observed. Urban-agricultural park is a better choice than urban park for wildlife. Thus, a higher number of preserved urban natural areas should be devoted to urban-agricultural parks. However, to increase the abundance of open-land species and in general wildlife, a less intensive management of cultivated and pasture patches is necessary. PMID- 11494073 TI - The value of sildenafil as mode of stimulation in pharmaco-penile duplex ultrasonography. AB - The purpose of this work was to assess whether a single intracavernous injection (ICI) of a low dose of the combination of papaverine-phentolamine is replaceable by a high dose of the oral erectogenic agent sildenafil as mode of stimulation during pharmaco-penile duplex ultrasonography (PPDU). Eleven patients with complaints of erectile dysfunction were included in a crossover study. With an interval of two weeks the patients were exposed to ICI with papaverine/phentolamine (3.75 mg/0.125 mg) and oral administration with sildenafil (100 mg) preceding PPDU. Five patients started with ICI. Six patients started with sildenafil. In the sildenafil stimulation mode, visual erotic stimulation (VES) was used to initiate erection. VES was applied by personal LCD monitor. Cut-off values to define sufficient arterial response were: peak flow velocity (PSV) >25 cm/s and acceleration time (AT) <72 ms. Cut-off value to define sufficient veno-occlusion was a resistance index > or =1.00. Statistical analysis of PPDU parameters shows no significant difference between the two modes of stimulation for arterial response (PSV, AT), whereas the resistance index, as a parameter of veno-occlusive response was significantly higher in the sildenafil mode. This finding is confirmed in the clinical translation of the results: two patients with an insufficient arterial response to ICI had a sufficient arterial response to sildenafil and only one patient showed an insufficient arterial response following sildenafil, whereas the response following ICI was sufficient. Analysis of veno-occlusive responses shows remarkable differences between both modes of stimulation. Whereas following the administration of sildenafil all veno occlusive responses were classified as sufficient, seven patients showed an insufficient veno-occlusive response following ICI. As mode of stimulation in PPDU, high dose sildenafil yields significantly less false positive diagnoses of 'veno-occlusive dysfunction' than intracavernous injection of the combination papaverine/phentolamine. No difference was found in the quality of the arterial response. Based on this study we conclude that sildenafil may replace ICI as mode of stimulation during PPDU. PMID- 11494074 TI - The efficacy and tolerability of vardenafil, a new, oral, selective phosphodiesterase type 5 inhibitor, in patients with erectile dysfunction: the first at-home clinical trial. AB - Vardenafil, a novel selective phosphodiesterase type 5 inhibitor, was evaluated in its first large-scale at-home trial. A total of 601 men with mild to severe erectile dysfunction (ED) were enrolled in this multi-centre, randomized, double blind, placebo-controlled trial of 12 weeks of treatment with either placebo or 5, 10 and 20 mg of vardenafil. Primary endpoints were Q3 (vaginal penetration) and Q4 (maintenance of erection) of the International Index of Erectile Function (IIEF). In the intent-to-treat population (n=580), the changes from baseline for 5, 10 and 20 mg vardenafil (1.2, 1.3 and 1.5, respectively) were all improved (P<0.001) over placebo (0.2) for Q3 and were similarly improved for Q4 (1.4, 1.5 and 1.7) compared to placebo (0.5) (P<0.001). All vardenafil doses improved all IIEF domains compared to placebo (P<0.001). The percentage of successful intercourses was between 71 and 75% for the three vardenafil doses. For the 20 mg dose, 80% of the patients experienced improved erections (GAQ) compared to 30% for placebo. Most frequent treatment-emergent adverse events were headache (7 15%), flushing (10-11%) and up to 7% for dyspepsia or rhinitis. Vardenafil treatment resulted in a high efficacy and low adverse-event profile in a population with mixed ED etiologies. PMID- 11494075 TI - Is there a role of radial rigidity in the evaluation of erectile dysfunction? AB - RigiScan has been the most widely utilized device for measuring erectile rigidity. However, the use of the RigiScan in the evaluation of erectile dysfunction has questionable because the RigiScan device does not directly determine axial rigidity. The aim of this study is to clarify that radial rigidity measured by RigiScan reflects the intracorporeal pressure and erectile capability efficiently. From January 1998 to May 1999, a total of 23 patients with erectile dysfunction were involved in the study. They were evaluated by RigiScan and duplex ultrasonography after intracorporeal injection of prostaglandin E1. We investigated the relationship between radial rigidity and the resistance index. The results of radial rigidity were also compared with that of the degree of erection. For the entire group, significant correlations were found between radial rigidity and the resistance index (r=0.680, P<0.001 for tip rigidity; r=0.703, P<0.001 for base rigidity). In addition, for 12 patients whose tip rigidity exceeded 60% and for 10 whose base rigidity exceeded 60%, the correlations between radial rigidity and the resistance index remained (r=0.659, P=0.020 for tip rigidity; r=0.759, P=0.011 for base rigidity). Based on the response determined by patients, radial rigidity represented the degree of erection efficiently. Our findings suggest that RigiScan is a useful diagnostic tool. Radial rigidity represents the intracorporeal pressure efficiently and has an acceptable role in the evaluation of erectile dysfunction. PMID- 11494077 TI - Effects of estrogen on nitric oxide synthase and histological composition in the rabbit clitoris and vagina. AB - We investigated the functional and histological changes after oophorectomy in the rabbit clitoris and vagina to determine the mechanism responsible for the development of arousal disorder in postmenopausal women. Twenty mature female New Zealand white rabbits were randomly divided into three groups: control; oophorectomy; and estrogen replacement after oophorectomy. We compared the nitric oxide synthase (NOS) activity and the degree of expression of neuronal (nNOS) and endothelial NOS (eNOS) using biochemical and Western blot analysis in clitoral and vaginal tissues. Histological change of smooth muscle and collagen contents in those tissues were also compared using Masson's trichrome staining. NOS activity and the expression of nNOS and eNOS were significantly increased in the oophorectomized group while there was a decrease to the level of the control group in the estrogen replacement group. Histological examination showed that oophorectomy induced a significant increase in collagen and decrease in muscle content in both clitoris and vagina, while the ratio of smooth muscle content was increased significantly after the estrogen replacement. Our results clearly demonstrate that estrogen deficiency induces compensatory NOS production which may be related to decreases in muscle to collagen ratio in female rabbit genital organs. PMID- 11494078 TI - Erection and NO override the vasoconstrictive effect of alpha-adrenergic stimulation in the rat penile vasculature. AB - Studies in this laboratory are designed to determine the effects of vasoconstrictor agents on the erectile response in rats. We have previously demonstrated that the vasoconstrictor effect of endothelin-1 (ET-1) is sharply reduced by erection and by nitric oxide (NO) administration. The present study was performed to determine if vasoconstriction, resulting from alpha-adrenergic stimulation, is altered by erection and NO. During continuous monitoring of corpus cavernosum pressure (CCP) and mean arterial pressure (MAP), erection was induced by electrical stimulation of the autonomic ganglion for the innervation of the penis. When the alpha-adrenergic agonist methoxamine (METH, 10 microg/kg) was injected before erection (ie, into the non-erect penis), the subsequent erectile response (CCP/MAP) was significantly reduced from 0.68+/-0.03 before METH to 0.34+/-0.08 after METH. Injection of METH into the erect penis (ie, during erection) reduced the vasoconstrictor action of METH; CCP/MAP was 0.74+/ 0.02 before METH and 0.55+/-0.05 after METH (P<0.05). The vasoconstrictor action of METH was slightly reduced when given in conjunction with NOR-1, a NO donor drug; CCP/MAP was 0.70+/-0.05 before METH, 0.55+/-0.09 after METH but this change was not significant. These results demonstrate that the response to alpha adrenergic stimulation is attenuated during erection in response to ganglionic stimulation. Furthermore, it appears that NO, produced during erection, may serve to override agonist-induced vasoconstriction. These results support our hypothesis that NO acts to directly stimulate relaxation of cavernous smooth muscle and to inhibit the vasoconstrictor actions of agents like ET-1 and alpha adrenergic agonists including norepinephrine. PMID- 11494079 TI - ASSESS-3: a randomised, double-blind, flexible-dose clinical trial of the efficacy and safety of oral sildenafil in the treatment of men with erectile dysfunction in Taiwan. AB - The efficacy and safety of sildenafil were evaluated in a randomised, double blind, placebo-controlled, flexible-dose study in Taiwanese men aged 26 to 80 y with erectile dysfunction (ED) of broad-spectrum aetiology and more than 6 months' duration. A total of 236 patients were randomised at six medical centres in Taiwan to receive either sildenafil (50 mg initially increased if necessary to 100 mg or decreased to 25 mg depending on efficacy and toleration) (n=119) or matching placebo (n=117) taken on an 'as needed' basis 1 h prior to anticipated sexual activity for a period of 12 weeks. At the end of 12 weeks, the primary efficacy variables relating to the achievement and maintenance of erections sufficient for sexual intercourse, and the secondary efficacy variables, which included: (1) the five separate domains of sexual functioning of the IIEF (International Index of Erectile Function) scale, (2) the percentage of successful intercourse attempts; and (3) a global assessment of erections, were all statistically significantly improved by sildenafil in comparison with placebo (P<0.0001). Treatment-related adverse events occurred in 43.7% of patients receiving sildenafil and 18.8% receiving placebo. The most common adverse events with sildenafil were flushing, dizziness and headache (25.2, 6.7 and 5.9% of patients, respectively), and most were mild in nature. The efficacy and safety of sildenafil in the population of Taiwanese men appears similar to that reported in other studies in western populations. PMID- 11494080 TI - A conscious-rabbit model to study vardenafil hydrochloride and other agents that influence penile erection. AB - Experimental models to study the effect of agents on penile erection usually include electrical stimulation of peripheral nerves in anesthetized animals combined with systemic or intracavernous injection of drugs. The objective of this study was to demonstrate that conscious rabbits can be used as a simple and quantitative model for the assessment of compounds that show potential for the treatment of erectile dysfunction. Erection was assessed by measuring the length of uncovered penile mucosa before and after the intravenous (i.v.) administration of agents. Animals did not require anesthesia during the course of the study. The phosphodiesterase 5 (PDE5) inhibitors vardenafil x HCl (hereafter called vardenafil) and sildenafil were given intravenously, and measurements were taken for 0-5 h. The effects of phentolamine and milrinone were also evaluated. Vardenafil (0.1-3 mg/kg) induced dose-dependent penile erections in conscious rabbits following i.v. administration. The efficacy of vardenafil was potentiated, and the minimal effective dose was reduced significantly to 0.01 mg/kg by simultaneous administration of the nitric oxide (NO) donor sodium nitroprusside (SNP). Administration of the NO-synthase inhibitor L-NAME abolished the effect. Sildenafil was effective in this model after i.v. administration. The alpha-adrenergic receptor antagonist phentolamine (0.1, 0.3 and 1 mg/kg i.v.) induced erections with a slower t(max) compared with vardenafil and sildenafil. Intravenous administration of the PDE3 inhibitor milrinone (1 mg/kg i.v.) was less effective than the PDE5 inhibitor vardenafil. The conscious rabbit is a suitable and reliable model for the evaluation of compounds with potential for the treatment of erectile dysfunction. This was demonstrated using compounds that target different signaling pathways that induce smooth muscle relaxation in the penis. PMID- 11494081 TI - Development of a mathematical model for the prediction of the area of venous leak. AB - Erectile dysfunction (ED) is an increasingly prevalent medical problem, affecting up to 50% of men aged between 40 and 70-y-old. Many cases are vasculogenic and some of these stem from the inability of the penis to store blood during erection due to leak into the venous system, termed corporo-venocclusive dysfunction (CVOD). The area of leakage during erection could be the most direct measure of erectile function but has not been investigated before. We have developed a simple mathematical model to determine the area of leak during erection and have tested it on data from both normal men (n=3) and men with venogenic impotence (n=16) undergoing dynamic infusion cavernosometry (DIC). The area of leak in the impotent group is significantly greater than in normal men at intracorporal pressures above 30 mmHg and reaches a plateau between 60 and 90 mmHg. Based on this study, we suggest that it may be necessary only to perform DIC at intracorporal pressures between 60 and 90 mmHg. PMID- 11494083 TI - Ferula harmonis 'zallouh' and enhancing erectile function in rats: efficacy and toxicity study. AB - Ferula harmonis, which is locally called 'zallouh' in the Middle East, is used as an aphrodisiac as it is reputed to enhance male sexual behavior, however, there is no scientific verification. In this study, the oil extracted from the seeds of Ferula harmonis was tested for its efficacy in enhancing erectile function and toxicity in male rats. The sexual activities assessed by penile erection index were dose dependent. The ED(50) (12.03 mg/kg) was 880 times less than the LD(50) (10.6 g/kg). However, when doses ranging from 0.05, 0.5 to 2 g/kg were given daily for 28 days, acute and subacute toxicity were observed. There was a decrease in total body weight, hepatomegaly, atrophic testis, significant decrease in hemoglobin and red blood cell count. In addition, there was a significant decrease in cholesterol level. All the above indicate that the crude oil from the plant Ferula harmonis can enhance erectile function, however, it becomes toxic if it is used for a long period of time. Further studies are underway to isolate and identify the active ingredients and their exact mechanisms of action. PMID- 11494082 TI - Dual mechanism of action of nicorandil on rabbit corpus cavernosal smooth muscle tone. AB - The potential of ATP-sensitive potassium channel openers (KCOs) for the treatment of male erectile dysfunction has recently been suggested based on positive clinical outcomes following intra-cavernosal administration of pinacidil. Agents that increase the levels of cGMP via elevation of nitric oxide (NO) nitroglycerin, for example, are also effective in improving erectile function preclinically and clinically. The aim of the present study was to determine the effects and mechanism of the action of nicorandil on rabbit corpus cavernosum. The in vitro regulation of smooth muscle tone was assessed in isolated cavernosal tissues pre-contracted with phenylephrine. Nicorandil, but not its major metabolite, relaxed phenylephrine-precontracted cavernosum smooth muscle with an EC(50) of 15 microM. The effects of nicorandil were only partially reversed by the K(ATP) channel blocker glyburide (10 microM) or by a soluble guanylate cyclase (sGC) inhibitor 1H-[1,2,4] oxadiazole [4,3-a] quinoxalin-1-one (ODQ, 3 microM). However, a combination of ODQ and glyburide completely blocked the relaxant effects of nicorandil. The results of the present study indicate that nicorandil can relax rabbit cavernosal tissue in vitro via a mechanism that involves activation of K(ATP) channels and stimulation of soluble guanylate cyclase. PMID- 11494085 TI - Assessment of as needed use of pharmacotherapy and the pause-squeeze technique in premature ejaculation by Abdel-Hamid et al. PMID- 11494086 TI - Community pharmacies for detection and control of hypertension. PMID- 11494087 TI - Menopause and the characteristics of the large arteries in a population study. AB - In previous cross-sectional and longitudinal population studies, we found that the slope of systolic pressure on age was steeper in postmenopausal than in premenopausal women. We hypothesised that this observation could be due to a specific effect of menopause on the elasticity of the large arteries. We investigated 315 randomly selected women, aged 30 to 70 years. Based on 5.2 years of follow-up, 166 women were premenopausal and 149 menopausal (44 reaching menopause and 105 postmenopausal). These women were matched on age and body mass index with 315 men. We used a wall-tracking ultrasound system to measure the diameter, compliance and distensibility of the brachial and the common carotid and femoral arteries as well as carotid-femoral pulse wave velocity. Pulse pressure was determined from 24-h blood pressure recordings. Both in menopausal women (r = 0.37; P < 0.001) and in matching male controls (r = 0.16; P = 0.04), pulse pressure widened with increasing age. The slope of the 24-h pulse pressure on age was steeper in menopausal women than in their premenopausal counterparts (0.428 vs -0.066 mm Hg per year; P = 0.003) and than in the male controls (0.428 vs 0.188 mm Hg per year; P = 0.06). After adjustment for age, 24-h mean pressure, body mass index, antihypertensive drug treatment, smoking and the use of oral contraceptives or hormonal replacement therapy, postmenopausal women showed a higher carotid-femoral pulse wave velocity (7.77 vs 6.71 m/s; P = 0.02) and had a slightly greater diameter of the common carotid artery (7.09 vs 6.79 mm; P = 0.07) than their premenopausal counterparts. After similar adjustments, menopausal class was not significantly associated with other vascular measurements in women or with any vascular measurement in control men. In conclusion, menopause per se may increase aortic stiffness. We hypothesise that this phenomenon may contribute to the rise in systolic pressure and pulse pressure in women beyond age 50 and, in turn, may lead to a slight dilatation of the common carotid artery. PMID- 11494088 TI - A study of inter-arm blood pressure differences in primary care. AB - OBJECTIVES: To investigate whether there are inter-arm blood pressure differences that are of clinical importance to general practice. DESIGN AND SETTING: Pragmatic study with randomised order of use of left or right arm carried out in routine surgeries in an inner city and suburban general practice. SUBJECTS: There were 237 patients presenting opportunistically for blood pressure measurement to a nurse or general practitioner. MAIN OUTCOME MEASURES: 95% limits of agreement between measurements on the left and right arm and bias between arms. RESULTS: Large inter-arm blood pressure differences exist reflected in wide 95% limits of agreement; -16 mm Hg to 24 mm Hg for the right minus the left arm diastolic blood pressure. There is a small but statistically significant bias to the right arm blood pressure measuring higher than the left (3.7 mm Hg diastolic, 2.4 to 5 mm Hg 95% confidence intervals). An interarm difference of 10 mm Hg or greater for diastolic blood pressure occurred in 40% of subjects and a difference of 20 mm Hg or more for systolic blood pressure occurred in 23% of subjects. CONCLUSIONS: In a primary care setting blood pressure should be measured routinely in both arms. If one arm is to be preferred for pragmatic clinical purposes, then this should be the right arm. PMID- 11494089 TI - Platelet sodium-hydrogen exchanger activity and left ventricular mass. AB - INTRODUCTION: The sodium-hydrogen exchanger (NHE) is integral to the processes that facilitate cell growth and may contribute to the development of left ventricular hypertrophy. The aim of this study was to examine the relationship between platelet sodium-hydrogen exchanger activity and left ventricular mass index (LVMI). METHODS: Twenty male untreated Caucasians (mean age +/- s.d.: 48 +/ 13; body mass index: 29 +/- 4 kg/m(2)) with a wide range of blood pressures were studied (mean BP: 152 +/- 22/93 +/- 15 mm Hg; range: 115-190/61-117 mm Hg). Sodium-hydrogen exchanger activity was determined as the rate of sodium-dependent recovery of intracellular pH in isolated platelets loaded with BCECF and acidified to pH 6.25 using nigericin. LVMI was calculated from measurements made by M-mode echocardiography. Associations between continuous variables were examined using parametric tests. RESULTS: The mean rate of pHi recovery was 0.15 +/- 0.03 dpHi/s (range: 0.09-0.21). Mean LVMI was 120 +/- 32 g/m(2) (range: 56 178) and was not significantly correlated with either systolic (r = 0.39, P = 0.09) or diastolic blood pressure (r = 0.27, P = 0.3). Platelet NHE activity was not significantly correlated with LVMI (r = 0.06; P = 0.8). Platelet NHE activity was not significantly different between subjects with (n = 7 with LVMI >131 g/m(2)) and without left ventricular hypertrophy (n = 13). CONCLUSIONS: The results of this study show that platelet NHE activity is not significantly correlated with LVMI. These data contrast with previously described correlations of LVMI with exchanger activity measured in leucocytes and erythrocytes; and indicate that the relationship between LVMI and exchanger activity may be specific to the cell type in which exchanger activity is measured. PMID- 11494090 TI - A physician-pharmacist model for the surveillance of blood pressure in the community: a feasibility study. AB - Hypertension is poorly managed. Hospital-based pharmacists working with physicians have been shown to improve the rate of achievement of "target" blood pressure in selected patients. It is unknown if such schemes can operate in the community and to what extent they would attract volunteers with poorly managed blood pressure. We assessed the feasibility of pharmacists to provide community based, open-access, blood pressure monitoring. In addition, we describe the blood pressure profile of the group in comparison to that of the 1994 Health Survey of England (HSE). Pharmacists from six pharmacies were trained to deliver the service. Adults living within the postal districts of the pharmacies were invited, through an advertising campaign, to volunteer to have their blood pressure measured. Blood pressure data and information on treatment for hypertension and/or diabetes were collected on 263 registrants. Patients were advised to have their blood pressure managed by the general practitioner immediately (category 1), re-measured within 2-3 months (category 2) or in 12 months time (category 3). The mean (s.d.) blood pressure of patients in categories 1 (n = 16), 2 (n = 117) and 3 (n = 130) was 186(16)/97(29), 151(13)/94(9) and 139(22)/86(13) mm Hg respectively; P < 0.001. Ninety-one patients (35%) were in receipt of antihypertensive therapy. Forty-five percent of the treated group had controlled blood pressure (<160/95 mm Hg) compared with 30% in the HSE dataset. A large proportion of known hypertensive patients with poor blood pressure control who had visited their general practitioner within the previous 6 months were detected by the pharmacist-led service. Pharmacists operating an open-access blood pressure monitoring service may be of value in improving the management of hypertension. PMID- 11494091 TI - The use of ambulatory blood pressure monitoring in managing hypertension according to different treatment guidelines. AB - OBJECTIVE: To investigate the use of ambulatory blood pressure monitoring (ABPM) in identifying and managing a group of patients referred to a tertiary centre for the assessment of their blood pressure and to illustrate the importance of introducing standardised ABPM treatment guidelines. PATIENTS AND METHODS: We examined 2000 sequential ABP recordings, 1557 were first time referrals from General Practitioners, Consultants and other hospitals. All patients were referred with suspected hypertension, resistant hypertension, white coat hypertension and for investigations of secondary hypertension. Fully trained nurse specialists fitted the monitors in the hypertension clinic and recordings were performed for 24 h. The data was then analysed and stratified according to treatment guidelines and categorised according to different definitions. RESULTS: The group of first time referrals (n = 1557) showed an even sex distribution of 789 men and 768 women, mean age 53 +/- 13.8 (12-88 years). Of this group 542 patients (35%) exhibited a white coat effect (WCE), 526 (34%) had a daytime ABP < or =139/89 mm Hg. Of these 81 (15%) had a high clinic blood pressure (ie, white coat hypertension (WCH)) according to our definition. Thirty-five of these patients were not on treatment but may have had it initiated on the basis of their clinic pressures. According to the British Hypertension Society (BHS) guidelines on clinic readings 772 (45%) of our patients would be classified as hypertensive or inadequately treated, 509 (33%) borderline and 326 (21%) as normal. Using daytime ABP levels according to O'Brien: 1031 (67%) would be defined as abnormal, 192 (12%) as borderline and 334 (21%) as normal. CONCLUSION: These results illustrates how patient management may differ markedly when treating in accordance either with the BHS guidelines for clinic readings or the suggested levels for ABP. More patients had abnormal blood pressure levels according to ABPM, even though it is superior in detecting WCE and WCH. Clear guidelines for ABPM treatment levels need to be established. PMID- 11494092 TI - Perceived symptoms and health-related quality of life reported by uncomplicated hypertensive patients compared to normal controls. AB - OBJECTIVES: This study compared the prevalence and intensity of symptoms and the health-related quality of life (HQL) of patients taking antihypertensive medications and patients without disease. METHODS: This cross-sectional study used surveys mailed to patient's homes. All consecutive patients over age 30 years attending either a general medicine or hypertension clinic during 3 months were eligible (n = 437). Hypertension group (HTN-G) patients were diagnosed with primary hypertension, prescribed antihypertensive medications, and had no other symptomatic conditions or drug therapies. Control group (CNTL-G) patients were seen in the general medicine clinic and had no chronic symptomatic conditions or drug therapies. Measures included the Symptom Distress Checklist (SDC, list of 51 symptoms, frequency, and level of distress), the Medical Outcomes Study Short Form-36 (SF-36), medications, blood pressures, and other data obtained from medical records and patient self-report. RESULTS: A total of 222 patients responded (46% CNTL-G, 55% HTN-G). HTN-G patients were somewhat older (59.0 +/- 11.2 vs 48.5 +/- 11.7 years, P = 0.001) and had a higher percent of minorities (24.8% vs 13.5%, P = 0.02), but otherwise similar. After adjusting for age and race differences, HTN-G patients reported significantly more symptoms (8.8 +/- 7.8 vs 4.7 +/- 4.8, P = 0.001) and related distress (32.2 +/- 4.2 vs 12.0 +/- 18.2, P = 0.001) as well as lower scores (reduced HQL) for most of the SF-36 domains. In general, hypertensive patients had more physical, but not mental symptoms than control patients. CONCLUSIONS: Hypertensive patients receiving antihypertensive medications have more symptoms and lower HQL. Differences were detected by both a brief, general HQL instrument and a detailed, disease-specific instrument. Routine screening of treated hypertensive patients using a brief HQL questionnaire to detect physical symptoms may prove feasible and useful. PMID- 11494093 TI - Liquorice-induced rise in blood pressure: a linear dose-response relationship. AB - To clarify the dose-response and the time-response relationship between liquorice consumption and rise in blood pressure and explore the inter-individual variance this intervention study was designed and executed in research laboratories at University hospitals in Iceland and Sweden. Healthy, Caucasian volunteers who also served as a control for himself/herself consumed liquorice in various doses, 50-200 g/day, for 2-4 weeks, corresponding to a daily intake of 75-540 mg glycyrrhetinic acid, the active substance in liquorice. Blood pressure was measured before, during and after liquorice consumption. Systolic blood pressure increased by 3.1-14.4 mm Hg (P < 0.05 for all), demonstrating a dose-response but not a time-response relationship. The individual response to liquorice followed the normal distribution. Since liquorice raised the blood pressure with a linear dose-response relationship, even doses as low as 50 g of liquorice (75 mg glycyrrhetinic acid) consumed daily for 2 weeks can cause a significant rise in blood pressure. The finding of a maximal effect of liquorice after only 2 weeks has important implications for all doctors dealing with hypertension. There does not seem to be a special group of responders since the degree of individual response to liquorice consumption followed the normal distribution curve. PMID- 11494094 TI - Polymorphism in exon 6 of the dopamine D(2) receptor gene (DRD2) is associated with elevated blood pressure and personality disorders in men. AB - A deficient dopamine D(2) receptor (DA2) formation or action may contribute to hypertension via an increase of catecholamine release. In addition, Axis II personality disorders that appears odd or eccentric (cluster A) is associated with a low density of DA2. This study sought to examine if a NcoI restriction fragment length polymorphism (C to T transition) in exon 6 of the dopamine D(2) receptor gene (DRD2) was associated with these characteristics. The genotypes (CC, CT and TT) were compared in anthropometric, endocrine, metabolic and haemodynamic variables as well as estimates of personality disorders in 284 randomly selected 51-year-old men. Homozygotes for the C allele constituted 49% of the men and homozygotes for the T allele 9%, while heterozygotes were 41%. The TT genotype was associated with elevated systolic and diastolic blood pressure, independent of obesity and endocrine abnormalities, including the hypothalamic pituitary-adrenal axis regulation. Moreover, the TT genotype was significantly more frequent among subjects with grade 1 (mild) hypertension (>140/90 mm Hg) compared to normotensive subjects (<130/85 mm Hg). The polymorphism in exon 6 of the DRD2 was also significantly associated with cluster A personality disorders. These results suggest that a polymorphism in exon 6 of the DRD2examined with the restriction enzyme NcoI is associated with an elevated blood pressure, independent of obesity. Paranoid or schizoid personality disorders is also associated with a polymorphism of the DRD2, which might be associated with a previously demonstrated low density of this receptor. PMID- 11494095 TI - Efficacy and safety of a therapeutic interchange from high-dose calcium channel blockers to a fixed-dose combination of amlodipine/benazepril in patients with moderate-to-severe hypertension. AB - BACKGROUND: Recent hypertension trials have demonstrated the importance of achieving goal blood pressures to reduce the risk of target organ damage. In patients with moderate to severe hypertension, the use of high-dose monotherapy and/or combinations of drugs are necessary to achieve these goals. Fixed-dose combination products may be useful in these patients by reducing the number of daily doses required to control blood pressure. OBJECTIVE: The objective of the present study was to evaluate the efficacy and safety of a therapeutic interchange between high-dose calcium channel blocker therapy and a fixed-dose combination of amlodipine/ benazepril (Lotrel; Novartis Pharmaceuticals, USA) in patients with moderate to severe hypertension. METHODS: A total of 75 patients were switched from amlodipine (n = 25), felodipine (n = 25), and nifedipine-GITS (n = 25) to amlodipine/benazepril. Twenty-eight of the 75 patients (37%) were taking either a beta-blocker or a diuretic in addition to the high-dose calcium channel blocker prior to the switch. Blood pressure control, side effects and the cost of the therapeutic interchange were evaluated in the year following the therapeutic interchange. RESULTS: Sixty-six of the 75 (88%) patients were successfully switched with maintenance of blood pressure control and without the development of new dose-limiting side effects. Reasons for treatment failure after the therapeutic interchange included loss of blood pressure control in five patients and the development of new dose-limiting side effects in four patients. These side effects included cough in three patients and rash in one patient. After accounting for differences in drug acquisition cost and costs related to the switch (clinic and emergency room and laboratory tests), a cost savings of $16030 for all 75 patients was realised in the first year. The per patient-per year cost savings was $214. CONCLUSIONS: Our data indicate that a therapeutic interchange from selected high-dose calcium channel blockers to a fixed-dose combination of amlodipine/ benazepril can be successfully accomplished in the majority of patients. PMID- 11494096 TI - Post exercise hypotension is sustained during subsequent bouts of mild exercise and simulated activities of daily living. AB - Our purpose was to examine whether the transient suppression of blood pressure that occurs during the hours following acute exercise (termed post exercise hypotension) persists throughout an active period of subsequent mild exercise and simulated activities of daily living (ADL) using direct measurements of arterial pressure. Eight recreationally active participants, with low borderline systolic hypertension completed 30 min of cycle ergometry at 70% VO(2Peak) and 30 min of quiet seated rest on separate days (randomised order). Following exercise and rest, subjects completed a 70-min protocol of mild exercise and simulated ADL. Blood pressure was monitored throughout by catheterisation of the radial artery. Exercise resulted in lower systolic (SBP), diastolic (DBP) and mean arterial pressure (MAP) throughout the post exercise ADL period compared to control measurements taken without prior exercise. The maximal difference in SBP, DBP and MAP between trials was 26, 7 and 13 mm Hg respectively. Average differences in SBP, DBP and MAP between trials were 16, 5 and 8 mm Hg respectively. This relative hypotension occurred in spite of higher heart rates during the ADL measurement period following the prior exercise. Furthermore, many of the blood pressure measurements during the post exercise period were significantly lower than the pre-exercise values during the same trial. We conclude that post exercise hypotension persists during mild exercise and simulated ADL. Although the duration of this relative hypotension needs to be determined, acute exercise may serve as a non-pharmacological aid in the treatment of hypertension. PMID- 11494097 TI - Hypertension is not a disease of the left arm: a difficult diagnosis of hypertension in Takayasu's arteritis. AB - Hypertension and its cause may be missed by failure to measure blood pressure in both arms. We report a case of Takayasu's arteritis where diagnostic confusion arose because there was a failure to detect a difference in blood pressure between the arms. PMID- 11494098 TI - Alterations of blood pressure and heart rate circadian rhythmic structure in non blind patients affected by retinitis pigmentosa. PMID- 11494099 TI - Microbial reduction of alpha-chloroketone to alpha-chlorohydrin. AB - Microbial reduction of alpha-chloroketone to alpha-chlorohydrin was studied as one of the approaches for construction of the chiral center of the corresponding epoxide. About 100 microorganisms covering many species of Candida, Pichia, Hansenula, Geotrichum, Rhodococcus and Aureobasidium were screened to reduce the alpha-chloroketone stereospecifically. Many strains provided the R-alpha chlorohydrin with 100% enantiomeric excess (ee), e.g., Candida sonorensis SC 16117, Geotrichum candidum SC 5469, Rhodotorula glutinis SC 16293, Sphingomonas paucimobilis SC 16113, Pichia silvicola SC 16159 and Rhodococcus equi SC 15835. Few microorganisms showed preferential formation of S-alpha-chlorohydrin after reduction. Among them, Pichia pinus SC 13864 and two Pichia methanolica strains SC 16116 and SC 13860 were the best, providing the S-alpha-chlorohydrin with ee of 88%, 79% and 78%, respectively. The enantiospecificity of the reduction by these Pichia species can be modified by changing the pH or prior heat treatment of the cells and S-alpha-chlorohydrin with > or =95% ee was obtained by appropriate modification of reaction conditions. PMID- 11494100 TI - Dromedary milk lactic acid fermentation: microbiological and rheological characteristics. AB - The ability of dromedary skim milk to form an acid curd during a lactic acid starter fermentation was investigated. The activity of the starter in dromedary milk was characterized by a longer lag phase ( approximately 5 vs. approximately 1 h) and by an earlier decline phase. This suggests the presence of inhibiting factors. The maximum buffering capacity of dromedary milk as well as its minimum apparent viscosity were obtained at lower pH values. Similarly, its elastic modulus appeared later (pH 5.7 vs. 6.3). Because these rheological and biochemical events took place at lower pH values, dromedary skim milk seems to present a higher physical stability toward the increase of acidity. Determination of the rheological and microscopic characteristics of the dromedary milk coagulum (pH 4.4) did not reveal curd formation but indicated a fragile and heterogeneous structure. This coagulum, which is very different from that of cows' milk, seems to be made up of dispersed casein flakes. PMID- 11494101 TI - Exopectinases produced by Aspergillus niger in solid-state and submerged fermentation: a comparative study. AB - Exopectinase production by Aspergillus niger was compared in submerged fermentation (SmF) and solid-state fermentation (SSF). SSF was carried out using polyurethane foam (PUF) as the solid support. The purpose was to study the effect of sucrose addition (0 or 40 g/l) and water activity level (A(w)=0.99 or 0.96) on the level of enzyme activity induced by 15 g/l of pectin. Mycelial growth, as well as extracellular protease production, was also monitored. Sucrose addition in SmF resulted in catabolite repression of exopectinase activity. However, in SSF, an enhancement of enzyme activity was observed. Protease levels were minimal in SSF experiments with sucrose and maximal in SmF without sucrose. Exopectinase yields (IU/g X) were negligible in SmF with sucrose. The high levels of exopectinase with sucrose and high A(w) in SSF can be explained by a much higher level of biomass production without catabolite repression and with lower protease contamination. PMID- 11494102 TI - Purification and properties of the cell-associated beta-xylosidase from Aureobasidium. AB - beta-Xylosidase was extracted from Aureobasidium sp. ATCC 20524 and purified to homogeneity. The molecular mass was estimated at 411 kDa. The enzyme contained 15.3% (w/w) carbohydrate. The optimum pH and temperature were pH 3.5 and 80 degrees C, respectively. The enzyme was stable at pH 3.5-9 after 3 h and at 80 degrees C after 15 min. The Michaelis constant (K(m)) and maximum velocity (V(max)) toward p-nitrophenyl-beta-D-xyloside were 2.0 mmol l(-1) and 0.94 mmol min(-1) mg(-1) protein, respectively. The enzyme was inhibited strongly by mercury, lead, and copper ions. PMID- 11494103 TI - Anka and anka pigment production. AB - This study was conducted to determine the time-dependent changes of solid-state fermentation of rice with Monascus purpureus to produce anka and anka pigments. Growth of the fungus occurred prior to the synthesis of anka pigments. A steady increase in the yield of pigments occurred between the 5th and 15th days. After 15 days, growth of the fungus on rice substrate ceased and the yield of yellow anka pigments remained constant; however, orange anka pigments were reduced with a decreasing rate of 3.6 mg/g anka/day. PMID- 11494104 TI - Biodegradation of polycyclic aromatic hydrocarbons by Sphingomonas strains isolated from the terrestrial subsurface. AB - Several strains of Sphingomonas isolated from deep Atlantic coastal plain aquifers at the US Department of Energy Savannah River Site (SRS) near Aiken, SC were shown to degrade a variety of aromatic hydrocarbons in a liquid culture medium. Sphingomonas aromaticivorans strain B0695 was the most versatile of the five strains examined. This strain was able to degrade acenaphthene, anthracene, phenanthrene, 2,3-benzofluorene, 2-methylnaphthalene, 2,3-dimethylnaphthalene, and fluoranthene in the presence of 400 mg l(-1) Tween 80. Studies involving microcosms composed of aquifer sediments showed that S. aromaticivorans B0695 could degrade phenanthrene effectively in sterile sediment and could enhance the rate at which this compound was degraded in nonsterile sediment. These findings indicate that it may be feasible to carry out (or, at least, to enhance) in situ bioremediation of phenanthrene-contaminated soils and subsurface environments with S. aromaticivorans B0695. In contrast, strain B0695 was unable to degrade fluoranthene in microcosms containing aquifer sediments, even though it readily degraded this polynuclear aromatic hydrocarbon (PAH) in a defined liquid growth medium. PMID- 11494105 TI - Soy molasses as fermentation substrate for production of butanol using Clostridium beijerinckii BA101. AB - Spray-dried soy molasses (SDSM) contains the sugars dextrose, sucrose, fructose, pinitol, raffinose, verbascose, melibiose, and stachyose. Of the 746 g kg(-1) total sugars in SDSM, 434 g kg(-1) is fermentable using Clostridium beijerinckii BA101. SDSM was used to produce acetone, butanol, and ethanol (ABE) by C. beijerinckii BA101 in batch cultures. Using 80 g l(-1) SDSM, 10.7 g l(-1) ABE was produced in P2 medium. Higher concentrations of SDSM resulted in poor solvent production due to the presence of excessive salt and inhibitory components. C. beijerinckii BA101 in SDSM at 80 g l(-1) concentration produced 22.8 g l(-1) ABE when supplemented with 25.3 g l(-1) glucose. SDSM contains 57.4 g kg(-1) mineral ash and 2% tri-calcium phosphate. Tri-calcium phosphate up to 43.1 g l(-1) was not inhibitory and at a tri-calcium phosphate concentration of 28.8 g l(-1), the culture produced more solvents (30.1 g l(-1)) than the control experiment (23.8 g l(-1)). In contrast, sodium chloride was a strong inhibitor of C. beijerinckii BA101 cell growth. At a concentration of 10 g l(-1) sodium chloride, a maximum cell concentration of 0.6 g l(-1) was achieved compared to 1.7 g l(-1) in the control experiment. The effects of two salts on specific growth rate constant (mu) and specific rate of ABE production (nu) for C. beijerinckii BA101 were examined. PMID- 11494106 TI - Production of tannase by Aspergillus niger Aa-20 in submerged and solid-state fermentation: influence of glucose and tannic acid. AB - Tannase production by Aspergillus niger Aa-20 was studied in submerged (SmF) and solid-state (SSF) fermentation systems with different tannic acid and glucose concentrations. Tannase activity and productivity were at least 2.5 times higher in SSF than in SmF. Addition of high tannic acid concentrations increased total tannase activity in SSF, while in SmF it was decreased. In SmF, total tannase activity increased from 0.57 to 1.03 IU/mL, when the initial glucose concentration increased from 6.25 to 25 g/L, but a strong catabolite repression of tannase synthesis was observed in SmF when an initial glucose concentration of 50 g/L was used. In SSF, maximal values of total tannase activity decreased from 7.79 to 2.51 IU when the initial glucose concentration was increased from 6.25 to 200 g/L. Kinetic results on tannase production indicate that low tannase activity titers in SmF could be associated to an enzyme degradation process which is not present in SSF. Tannase titers produced by A. niger Aa-20 are fermentation system dependent, favoring SSF over SmF. PMID- 11494107 TI - Influence of pH and ionic strength on adhesion of a wild strain of Pseudomonas sp. to titanium. AB - The adherence of an environmental strain of Pseudomonas sp. to titanium was evaluated modifying the pH (2 to 8) and ionic strength (0.1 and 0.6 M NaCl) of the electrolyte solution. Results were analyzed considering the participation of the different interfacial forces under the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. At 0.1 M, maximal bacterial adhesion was at pH 6, in agreement with the point of zero charge of the titanium surface. Similar adhesion values were observed at both sides of this point despite the opposite electrostatic condition of the surface oxide. At 0.6 M an absence of bacterial adhesion was observed throughout the pH range tested. The changes in bacterial adhesion are in agreement with the changes in the number of reinforced H-bond-forming sites on the titanium surface calculated using a simple model for the ionization of OH group adsorbed to the surface. PMID- 11494108 TI - Conversion of gamma-butyrobetaine to L-carnitine by Achromobacter cycloclast. AB - L-Carnitine is an ubiquitous substance that plays a major role in the transportation of long-chain fatty acids. We investigated crucial factors that influence microbial conversion of gamma-butyrobetaine to L-carnitine using an Achromobacter cycloclast strain. Two-stage culture results showed that gamma butyrobetaine induced enzymes essential for the conversion, which suggests that the precursor should be present in the initial cell growth stage. The addition of yeast extract enhanced L-carnitine production whereas inorganic nitrogen sources inhibited it. Under nitrogen-limiting conditions, the cells accumulated poly-beta hydroxybutyrate instead of L-carnitine. Among the trace elements tested, nickel addition enhanced L-carnitine production by almost twice that of the control and copper strongly inhibited the conversion. L-Carnitine production was reduced when the medium contained inorganic salts of sodium, potassium, and calcium at a concentration greater than 2 g l(-1). A higher L-carnitine yield was achieved when cells were incubated in a lower culture volume. The optimal pH for L carnitine production was 5 to 5.5, whereas that of growth was 7.0, indicating that a pH shift was required. Under optimal conditions, L-carnitine concentrations as high as 15 g l(-1) were obtained in 62 h with a 45% molar conversion yield. PMID- 11494109 TI - Biodegradation of BTEX vapors in a silicone membrane bioreactor system. AB - The biotreatment of complex mixtures of volatile organic compounds (VOCs) such as benzene, toluene, ethylbenzene, and xylene isomers (BTEX) has been investigated by many workers. However, the majority of the work has dealt with the treatment of aqueous or soil phase contamination. The biological treatment of gas and vapor phase sources of VOC wastes has recently received attention with increased usage of biofilters and bioscrubbers. Although these systems are relatively inexpensive, performance problems associated with biomass plugging, gas channeling, and support media acidification have limited their adoption. In this report we describe the development and evaluation of an alternative biotreatment system that allows rapid diffusion of both BTEX and oxygen through a silicone membrane to an active biofilm. The bioreactor system has a rapid liquid recycle, which facilitates nutrient medium mixing over the biofilm and allows for removal of sloughing cell mass. The system removed BTEX at rates up to 30 microg h(-1) cm(-2) of membrane area. BTEX removal efficiencies ranged from 75% to 99% depending on the BTEX concentration and vapor flowrate. Consequently, the system can be used for continuous removal and destruction of BTEX and other potential target VOCs in vapor phase streams. PMID- 11494110 TI - Novel DNA copy number losses in chromosome 12q12--q13 in adenoid cystic carcinoma. AB - In order to find common genetic abnormalities that may identify loci of genes involved in the development of adenoid cystic carcinoma (ACC), we investigated DNA copy number changes in 24 of these tumors by comparative genomic hybridization (CGH). Our results indicate that unlike many carcinomas, ACCs have relatively few changes in DNA copy number overall. Twenty tumors had DNA copy number changes, which were mostly restricted to a few chromosomal arms. A frequent novel finding was the loss of DNA copy number in chromosome 12q (eight tumors, 33%) with the minimal common overlapping region at 12q12--q13. Deletion in this region has not been reported to be frequent in other types of cancer analyzed by CGH. In addition, deletions in 6q23-qter and 13q21--q22 and gains of chromosome 19 were observed in 25% to 38% of ACCs. Deletion of 19q, previously reported in a small series of ACC, was not identified in the current group of carcinomas. The current CGH results for chromosomes 12 and 19 were confirmed by microsatellite allelotyping. These results indicate that DNA copy number losses in 12q may be important in the oncogenesis of ACC and suggest that the 12q12--q13 region may harbor a new tumor-suppressor gene. PMID- 11494111 TI - Poly(ADP-ribosyl)ation of p53 in vitro and in vivo modulates binding to its DNA consensus sequence. AB - The tumor-suppressor p53 undergoes extensive poly(ADP-ribosyl)ation early during apoptosis in human osteosarcoma cells, and degradation of poly(ADP-ribose) (PAR) attached to p53 coincides with poly(ADP-ribose)polymerase-1, (PARP-1) cleavage, and expression of p53 target genes. The mechanism by which poly(ADP-ribosyl)ation may regulate p53 function has now been investigated. Purified wild-type PARP-1 catalyzed the poly(ADP-ribosyl) of full-length p53 in vitro. In gel supershift assays, poly(ADP-ribosyl)ation suppressed p53 binding to its DNA consensus sequence; however, when p53 remained unmodified in the presence of inactive mutant PARP-1, it retained sequence-specific DNA binding activity. Poly(ADP ribosyl)ation of p53 by PARP-1 during early apoptosis in osteosarcoma cells also inhibited p53 interaction with its DNA consensus sequence; thus, poly(ADP ribosyl)ation may represent a novel means for regulating transcriptional activation by p53 in vivo. PMID- 11494113 TI - Familial/bilateral and sporadic testicular germ cell tumors show frequent genetic changes at loci with suggestive linkage evidence. AB - Testicular germ cell tumor (TGCT) is the most common tumor type among adolescent and young adult males. Familial clustering and bilateral disease are suggestive of a genetic predisposition among a subgroup of these patients, but susceptibility genes for testicular cancer have not yet been identified. However, suggestive linkage between disease and genetic markers has been reported at loci on chromosome arms 3q, 5q, 12q, 18q, and Xq. We have analyzed primary familial/bilateral (n=20) and sporadic (n=27) TGCTs, including 28 seminomas and 19 nonseminomas, for allelic imbalance (AI) within the autosomal regions. DNA from all tumors were analyzed by fluorescent polymerase chain reaction of 22 polymorphic loci at 3q27-ter, 5q13-35.1, 12q21-ter, and 18q12--ter. All tumor genotypes were evaluated against their corresponding constitutional genotypes. The percentages of TGCTs with genetic changes at 3q, 5q, 12q, and 18q, were 79%, 36%, 53% and 43%, respectively. The frequencies at 3q and 12q in nonseminomas were significantly higher than in seminomas (P=.003 and P=.004). In order to evaluate changes at hemizygous Xq loci, five loci were analyzed by co amplification with an autosomal reference marker known to reveal retained heterozygosity in the tumor DNA. Gain of Xq sequences was seen in more than 50% of the tumors. The degree of amplification varied among the loci in each of five tumors, and based on these breakpoints, a common region of overlapping gains was found at Xq28. No significant differences were found between the frequencies of genetic changes in familial/bilateral versus sporadic tumors, an observation speaking in disfavor of the existence of a single susceptibility gene for TGCT in any of the analyzed regions. Our data suggest that gain of genetic material at distal Xq and losses at 5q and 18q contribute to establishment of seminomas, whereas imbalances at 3q as well as gain at distal part of 12q are associated with further progression into nonseminomas. PMID- 11494112 TI - Using positron emission tomography with [(18)F]FDG to predict tumor behavior in experimental colorectal cancer. AB - This study investigates the relationship between FDG uptake as determined by positron emission tomography (PET) imaging and rates of tumor growth, cellular GLUT1 transporter density, and the activities of hexokinase and glucose-6 phosphatase in a solid tumor implant model. Five different human colorectal xenografts of different growth properties were implanted in athymic rats and evaluated by dynamic (18)F-FDG-PET. The phosphorylating and dephosphorylating activities of the key glycolytic enzymes, hexokinase and glucose-6-phosphatase, were measured in these tumor types by spectrophotometric assays and the expression of GLUT1 glucose transporter protein was determined by immunohistochemistry. Correlations among FDG accumulation, hexokinase activity, and tumor doubling time are reported in these colon xenografts. The results indicate that the activity of tumor hexokinase may be a marker of tumor growth rate that can be determined by (18)F-FDG-PET imaging. PET scanning may not only be a useful tool for staging patients for extent of disease, but may provide important prognostic information concerning the proliferative rates of malignancies. PMID- 11494114 TI - Evaluation of breast cancer polyclonality by combined chromosome banding and comparative genomic hybridization analysis. AB - Cytogenetically unrelated clones have been detected by chromosome banding analysis in many breast carcinomas. Because these karyotypic studies were performed on short-term cultured samples, it may be argued that in vitro selection occurred or that small clones may have arisen during culturing. To address this issue, we analyzed 37 breast carcinomas by G-banding and comparative genomic hybridization (CGH), a fluorescent in situ hybridization--based screening technique that does not require culturing or tumor metaphases. All but two of the 37 karyotypically abnormal cases presented copy number changes by CGH. The picture of genomic alterations revealed by the two techniques overlapped only partly. Sometimes the CGH analysis revealed genomic imbalances that belonged to cell populations not picked up by the cytogenetic analysis and in other cases, especially when the karyotypes had many markers and chromosomes with additional material of unknown origin, CGH gave a more reliable overall picture of the copy number gains and losses. However, besides sometimes revealing cell populations with balanced chromosome aberrations or unbalanced changes that nevertheless remained undetected by CGH, G-banding analysis was essential to understand how the genomic imbalances arose in the many cases in which both techniques detected the same clonal abnormalities. Furthermore, because CGH pictures only imbalances present in a significant proportion of the test sample, the very detection by this technique of imbalances belonging to apparently small, cytogenetically unrelated clones of cells proves that these clones must have been present in vivo. This constitutes compelling evidence that the cytogenetic polyclonality observed after short-term culturing of breast carcinomas is not an artifact. PMID- 11494115 TI - The protrusive phase and full development of integrin-dependent adhesions in colon epithelial cells require FAK- and ERK-mediated actin spike formation: deregulation in cancer cells. AB - Integrins play an important role in tumour progression by influencing cellular responses and matrix-dependent adhesion. However, the regulation of matrix dependent adhesion assembly in epithelial cells is poorly understood. We have investigated the integrin and signalling requirements of cell-matrix adhesion assembly in colon carcinoma cells after plating on fibronectin. Adhesion assembly in these, and in the adenoma cells from which they were derived, was largely dependent on alpha v beta 6 integrin and required phosphorylation of FAK on tyrosine-397. The rate of fibronectin-induced adhesion assembly and the expression of both alpha v beta 6 integrin and FAK were increased during the adenoma-to-carcinoma transition. The matrix-dependent adhesion assembly process, particularly the final stages of complex protrusion that is required for optimal cell spreading, required the activity of extracellular signal-regulated kinase (ERK). Furthermore, phosphorylated ERK was targeted to newly forming cell--matrix adhesions in the carcinoma cells but not the adenoma cells, and inhibition of FAK -tyrosine-397 phosphorylation or MEK suppressed the appearance of phosphorylated ERK at peripheral sites. In addition, inhibition of MEK--ERK activation blocked the formation of peripheral actin microspikes that were necessary for the protrusive phase of cell-matrix adhesion assembly. Thus, MEK--ERK--dependent peripheral actin re-organization is required for the full development of integrin induced adhesions and this pathway is stimulated in an in vitro model of colon cancer progression. PMID- 11494116 TI - Acute metabolic alkalosis enhances response of C3H mouse mammary tumors to the weak base mitoxantrone. AB - Uptake of weak acid and weak base chemotherapeutic drugs by tumors is greatly influenced by the tumor extracellular/interstitial pH (pH(e)), the intracellular pH (pH(i)) maintained by the tumor cells, and by the ionization properties of the drug itself. The acid-outside plasmalemmal pH gradient in tumors acts to exclude weak base drugs like the anthracyclines, anthraquinones, and vinca alkaloids from the cells, leading to a substantial degree of "physiological drug resistance" in tumors. We have induced acute metabolic alkalosis in C3H tumor-bearing C3H/hen mice, by gavage and by intraperitoneal (i.p.) administration of NaHCO(3). (31)P magnetic resonance spectroscopic measurements of 3-aminopropylphosphonate show increases of up to 0.6 pH units in tumor pH(e), and 0.2 to 0.3 pH units in hind leg tissue pH(e), within 2 hours of i.p. administration of NaHCO(3). Theoretical calculations of mitoxantrone uptake into tumor and normal (hind leg) tissue at the measured pH(e) and pH(i) values indicate that a gain in therapeutic index of up to 3.3-fold is possible with NaHCO(3) pretreatment. Treatment of C3H tumor bearing mice with 12 mg/kg mitoxantrone resulted in a tumor growth delay of 9 days, whereas combined NaHCO(3)--mitoxantrone therapy resulted in an enhancement of the TGD to 16 days. PMID- 11494117 TI - Rapid mutation scanning of genes associated with familial cancer syndromes using denaturing high-performance liquid chromatography. AB - Germline mutations in tumor suppressor genes, or less frequently oncogenes, have been identified in up to 19 familial cancer syndromes including Li-Fraumeni syndrome, familial paraganglioma, familial adenomatous polyposis coli and breast and ovarian cancers. Multiple genes have been associated with some syndromes as approximately 26 genes have been linked to the development of these familial cancers. With this increased knowledge of the molecular determinants of familial cancer comes an equal expectation for efficient genetic screening programs. We have trialled denaturing high-performance liquid chromatography (dHPLC) as a tool for rapid germline mutation scanning of genes implicated in three familial cancer syndromes -- Cowden syndrome (PTEN mutation), multiple endocrine neoplasia type 2 (RET mutation) and von Hippel-Lindau disease (VHL mutation). Thirty-two mutations, including 21 in PTEN, 9 in RET plus a polymorphism, and 2 in VHL, were analyzed using the WAVE DNA fragment analysis system with 100% detection efficiency. In the case of the tumor suppressor gene PTEN, mutations were scattered along most of the gene. However, mutations in the RET proto-oncogene associated with multiple endocrine neoplasia type 2 were limited to specific clusters or "hot spots." The use of GC-clamped primers to scan for mutations scattered along PTEN exons was shown to greatly enhance the sensitivity of detection of mutant hetero- and homoduplex peaks at a single denaturation temperature compared to fragments generated using non--GC-clamped primers. Thus, when scanning tumor suppressor genes for germline mutation using dHPLC, the incorporation of appropriate GC-clamped primers will likely increase the efficiency of mutation detection. PMID- 11494118 TI - Chromosome 7 aneusomy. A marker for metastatic melanoma? Expression of the epidermal growth factor receptor gene and chromosome 7 aneusomy in nevi, primary malignant melanomas and metastases. AB - Receptor tyrosine kinases such as the epidermal growth factor receptor (EGFR) play an important role in a variety of malignant neoplasias, making the search for aberrations in the relevant chromosomes an important issue. Differential expression of the EGFR gene was investigated by reverse transcriptase (RT)-PCR on tissue samples of normal skin, nevi, primary melanomas, and melanoma metastases. The EGFR gene is located on chromosome 7p12.3-p12.1. To determine the number of chromosomes 7 in cell nuclei of the mentioned tissue samples we performed fluorescence in situ hybridization (FISH) on touch preparations, using a DNA probe that hybridizes specifically to the centromeric region of chromosome 7. Additionally, chromosome 7 number in interphase nuclei was determined in short term primary cell cultures of nevi, primary melanomas, and metastases. The highest EGFR gene expression frequency was found in melanoma metastases. By FISH we detected the highest fraction of cell nuclei with more than two chromosomes 7 in the group of metastases. Our results suggest that overexpression of the EGFR gene might play an important role in metastasis of malignant melanoma. This is well reflected by polysomy 7, possibly accounting for an increased EGFR gene copy number. PMID- 11494119 TI - A hypoxia-regulated adeno-associated virus vector for cancer-specific gene therapy. AB - The presence of hypoxic cells in human brain tumors is an important factor leading to resistance to radiation therapy. However, this physiological difference between normal tissues and tumors also provides the potential for designing cancer-specific gene therapy. We compared the increase of gene expression under anoxia (<0.01% oxygen) produced by 3, 6, and 9 copies of hypoxia responsive elements (HRE) from the erythropoietin gene (Epo), which are activated through the transcriptional complex hypoxia-inducible factor 1 (HIF-1). Under anoxic conditions, nine copies of HRE (9XHRE) yielded 27- to 37-fold of increased gene expression in U-251 MG and U-87 MG human brain tumor cell lines. Under the less hypoxic conditions of 0.3% and 1% oxygen, gene activation by 9XHRE increased expression 11- to 18-fold in these cell lines. To generate a recombinant adeno associated virus (rAAV) in which the transgene can be regulated by hypoxia, we inserted the DNA fragment containing 9XHRE and the LacZ reporter gene into an AAV vector. Under anoxic conditions, this vector produced 79- to 110-fold increase in gene expression. We believe this hypoxia-regulated rAAV vector will provide a useful delivery vehicle for cancer-specific gene therapy. PMID- 11494121 TI - Link of a new type of apoptosis-inducing gene ASY/Nogo-B to human cancer. AB - Although apoptosis plays an essential role in the embryogenesis and homeostasis of multicellular organisms, this mechanism has not yet been fully clarified. We isolated a novel human apoptosis-inducing gene, ASY, which encodes an endoplasmic reticulum-targeting protein without any known apoptosis-related motifs. This gene is identical to the Nogo-B, a splice variant of the Nogo-A which has recently been shown to be an inhibitor of neuronal regeneration in the central nervous system. Ectopic expression of the ASY gene led to extensive apoptosis, particularly in cancer cells. Furthermore, transcription of the ASY gene was suppressed in small cell lung cancer. These results suggest that a new type of apoptosis-inducing gene, namely, ASY, may be involved in the development of certain types of cancer. PMID- 11494122 TI - Development of mammary adenocarcinomas by tissue-specific knockout of Brca2 in mice. AB - Cre-mediated deletion of exons 3 and 4 of the mouse Brca2 gene occurring specifically in mammary epithelial cells of conditional female mutants carrying a combination of loxP-modified and null Brca2 alleles resulted in a high incidence (77%) of breast tumors that were often palpable and developed in one or more glands after long latency (time for median tumor-free survival of approximately 1.4 years; total of 40 tumors in 20 animals). These invasive adenocarcinomas were histologically quite uniform, exhibiting predominantly a solid, nodular tumor pattern with very few variants, in striking contrast to the morphological heterogeneity of analogous Brca1-associated tumors. The karyotypes of tumor cells lacking Brca2 had various chromosomal aberrations and ranged from diploid to hypertetraploid, but this wide variability was incongruous with the histological appearance of carcinomas that was comparable between specimens. The implications of these observations in the context of models positing that Brca2 is involved in the maintenance of genomic stability are discussed. PMID- 11494123 TI - Protein phosphatases 1 and 2A promote Raf-1 activation by regulating 14-3-3 interactions. AB - Raf-1 activation is a complex process which involves plasma membrane recruitment, phosphorylation, protein-protein and lipid-protein interactions. We now show that PP1 and PP2A serine-threonine phosphatases also have a positive role in Ras dependent Raf-1 activation. General serine-threonine phosphatase inhibitors such sodium fluoride, or ss-glycerophosphate and sodium pyrophosphate, or specific PP1 and PP2A inhibitors including microcystin-LR, protein phosphatase 2A inhibitor I(1) or protein phosphatase inhibitor 2 all abrogate H-Ras and K-Ras dependent Raf-1 activation in vitro. A critical Raf-1 target residue for PP1 and PP2A is S259. Serine phosphatase inhibitors block the dephosphorylation of S259, which accompanies Raf-1 activation, and Ras dependent activation of mutant Raf259A is relatively resistant to serine phosphatase inhibitors. Sucrose gradient analysis demonstrates that serine phosphatase inhibition increases the total amount of 14 3-3 and Raf-1 associated with the plasma membrane and significantly alters the distribution of 14-3-3 and Raf-1 across different plasma membrane microdomains. These observations suggest that dephosphorylation of S259 is a critical early step in Ras dependent Raf-1 activation which facilitates 14-3-3 displacement. Inhibition of PP1 and PP2A therefore causes plasma membrane accumulation of Raf 1/14-3-3 complexes which cannot be activated. PMID- 11494124 TI - The adapter protein, Grb10, is a positive regulator of vascular endothelial growth factor signaling. AB - Vascular endothelial growth factor (VEGF) is an important regulator of vasculogenesis and angiogenesis. Activation of VEGF receptors leads to the recruitment of SH2 containing proteins which link the receptors to the activation of signaling pathways. Here we report that Grb10, an adapter protein of which the biological role remains unknown, is tyrosine phosphorylated in response to VEGF in endothelial cells (HUVEC) and in 293 cells expressing the VEGF receptor KDR. An intact SH2 domain is required for Grb10 tyrosine phosphorylation in response to VEGF, and this phosphorylation is mediated in part through the activation of Src. In HUVEC, VEGF increases Grb10 mRNA level. Expression of Grb10 in HUVEC or in KDR expressing 293 cells results in an increase in the amount and in the tyrosine phosphorylation of KDR. In 293 cells, this is correlated with the activation of signaling molecules, such as MAP kinase. By expressing mutants of Grb10, we found that the positive action of Grb10 is independent of its SH2 domain. Moreover, these Grb10 effects on KDR seem to be specific since Grb10 has no effect on the insulin receptor, and Grb2, another adapter protein, does not mimic the effect of Grb10 on KDR. In conclusion, we propose that VEGF up regulates Grb10 level, which in turn increases KDR molecules, suggesting that Grb10 could be involved in a positive feedback loop in VEGF signaling. PMID- 11494125 TI - Spleen tyrosine kinase (Syk) deficiency in childhood pro-B cell acute lymphoblastic leukemia. AB - The cytoplasmic spleen tyrosine kinase (SYK) is a key regulator of signal transduction events, apoptosis and orderly cell cycle progression in B-lineage lymphoid cells. Although SYK has not been linked to a human disease, defective expression of the closely related T-cell tyrosine kinase ZAP-70 has been associated with severe combined immunodeficiency. Childhood CD19(+)CD10(-) pro-B cell acute lymphoblastic leukemia (ALL) is thought to originate from B-cell precursors with a maturational arrest at the pro-B cell stage and it is associated with poor prognosis. Since lethally irradiated mice reconstituted with SYK-deficient fetal liver-derived lymphohematopoietic progenitor cells show a block in B-cell ontogeny at the pro-B to pre-B cell transition, we examined the SYK expression profiles of primary leukemic cells from children with pro-B cell ALL. Here we report that leukemic cells from pediatric CD19(+)CD10(-) pro-B cell ALL patients (but not leukemic cells from patients with CD19(+)CD10(+) common pre pre-B cell ALL) have markedly reduced SYK activity. Sequencing of the reverse transcriptase-polymerase chain reaction (RT-PCR) products of the Syk mRNA in these pro-B leukemia cells revealed profoundly aberrant coding sequences with deletions or insertions. These mRNA species encode abnormal SYK proteins with a missing or truncated catalytic kinase domain. In contrast to pro-B leukemia cells, pre-pre-B leukemia cells from children with CD19(+)CD10(+) common B lineage ALL and EBV-transformed B-cell lines from healthy volunteers expressed wild-type Syk coding sequences. Examination of the genomic structure of the Syk gene by inter-exonic PCR and genomic cloning demonstrated that the deletions and insertions in the abnormal mRNA species of pro-B leukemia cells are caused by aberrant splicing resulting in either mis-splicing, exon skipping or inclusion of alternative exons, consistent with an abnormal posttranscriptional regulation of alternative splicing of Syk pre-mRNA. Our findings link for the first time specific molecular defects involving the Syk gene to an immunophenotypically distinct category of childhood ALL. To our knowledge, this is the first discovery of a specific tyrosine kinase deficiency in a human hematologic malignancy. PMID- 11494126 TI - Fine chromosomal localization of the mouse Par2 gene that confers resistance against urethane-induction of pulmonary adenomas. AB - BALB/cByJ mice are 14 times more resistant to urethane-induction of pulmonary adenomas than the susceptible A/J strain. Our previous linkage analysis of (A/J x BALB/cByJ)F1 x A/J backcross mice provided statistical evidence that a major resistance locus of BALB/cByJ with a dominant effect, designated Par2 (Pulmonary adenoma resistance 2), exists within an approximately 25 cM section of distal chromosome 18. To facilitate molecular identification of the Par2 locus, the present study was conducted to finely localize its chromosomal position utilizing Par2-congenic mice. Male BALB/cByJ mice were mated with female C57BL/6J mice carrying recessive Par2 alleles and their male F1 progeny were backcrossed to female BALB/cByJ mice. A male backcross mouse heterozygous within the Par2 interval of 25 cM was randomly selected and again backcrossed to female BALB/cByJ mice. This backcross-selection cycle was simply repeated to produce semi-congenic mice with a general BALB/cByJ genetic background except for the Par2 interval, where the mice were heterozygous with paternal C57BL/6J alleles and maternal BALB/cByJ alleles. After the 6th or 7th backcross, nine male mice possessing a recombination within the paternal Par2 interval were retained and crossed to female A/J mice. Resultant progeny were treated with urethane and examined for lung tumor development in order to deduce the Par2 genotypes of the recombinants through linkage analysis. By comparing the deduced Par2 genotype of each recombinant with its recombinational breakpoint, the Par2 locus was confined to an approximately 0.5 cM region flanked by D18Mit103 and D18Mit188 loci. Our results indicate that fully congenic mice conventionally established by at least nine simple backcrosses or by the speed congenic method are not necessarily required for fine mapping of quantitative trait loci. In the case of the Par2 locus, we found that semi-congenic mice after as few as four simple backcrosses were useful for this purpose. The map information obtained in this study should enable subsequent positional cloning of the Par2 gene. PMID- 11494127 TI - C-cell and thyroid epithelial tumours and altered follicular development in transgenic mice expressing the long isoform of MEN 2A RET. AB - Gain-of-function mutations in the gene encoding the receptor tyrosine kinase RET have been identified as the aetiological factor for multiple endocrine neoplasia type 2A (MEN2A). MEN2A is a dominantly-inherited cancer predisposition syndrome characterized by medullary thyroid carcinoma, a tumour of the calcitonin producing thyroid C-cells. There are three isoforms of RET: RET9, RET43 and RET51, and although in vitro evidence suggests they vary in cellular transformation activities, little is known about their function in tumorigenesis in vivo. To address this, we used RET51 cDNA to construct mice in which the most frequent MEN2A mutation, Cys-634-Arg, was expressed under the control of the human calcitonin promoter (CT-2A mice). These mice developed C-cell tumours resembling human MTC and follicular tumours resembling human papillary thyroid carcinoma (PTC) depending on the founder line examined. One founder line developed compound MTC/PTC at low frequency (8%) and pancreatic cystadenocarcinoma. CT-2A mice also displayed a developmental defect in thyroid follicular structure, in which much of the thyroid was occupied by large irregular cystic follicles thought to be derived from the ultimobranchial body, a developmental precursor of the thyroid gland. The CT-2A mice will provide a suitable model to further study the effects of the MEN 2A RET mutation in vivo. PMID- 11494128 TI - Multiple signaling interactions of Abl and Arg kinases with the EphB2 receptor. AB - The Eph family of receptor tyrosine kinases and the Abl family of non-receptor tyrosine kinases have both been implicated in tissue morphogenesis. They regulate the organization of the actin cytoskeleton in the developing nervous system and participate in signaling pathways involved in axon growth. Both Eph receptors and Abl are localized in the neuronal growth cone, suggesting that they play a role in axon pathfinding. Two-hybrid screens identified regions of Abl and Arg that bind to the EphB2 and EphA4 receptors, suggesting a novel signaling connection involving the two kinase families. The association of full-length Abl and Arg with EphB2 was confirmed by co-immunoprecipitation and found to involve several distinct protein interactions. The SH2 domains of Abl and Arg bind to tyrosine phosphorylated motifs in the juxtamembrane region of EphB2. A second, phosphorylation-independent interaction with EphB2 involves non-conserved sequences in the C-terminal tails of Abl and Arg. A third interaction between Abl and EphB2 is probably mediated by an intermediary protein because it requires tyrosine phosphorylation of EphB2, but not the binding sites for the Abl SH2 domain. The connection between EphB2 and Abl/Arg appears to be reciprocal. Activated EphB2 causes tyrosine phosphorylation of Abl and Arg, and vice versa. Interestingly, treatment of COS cells and B35 neuronal-like cells with ephrin-B1 to activate endogenous EphB2 decreased the kinase activity of endogenous Abl. These data are consistent with the opposite effects that Eph receptors and Abl have on neurite ougrowth and suggest that Eph receptors and Abl family kinases have shared signaling activities. PMID- 11494129 TI - Change in gene expression subsequent to induction of Pnn/DRS/memA: increase in p21(cip1/waf1). AB - Pnn (PNN) is a nuclear and cell adhesion-related protein. Previous work has suggested that Pnn/DRS/memA is a potential tumor suppressor involved in the regulation of cell adhesion and cell migration. Using the ecdysone-inducible mammalian expression system, a stable inducible GFP-tagged human Pnn gene (PNNGFP) expressing 293 cell line was created (EcR293-PNNGFP). Cells induced to express PNNGFP not only exhibited increased cell-cell adhesion but also exhibited changes in cell growth and cell cycle progression. cDNA array analyses, together with real time PCR, revealed that the effects of exogenously expressed Pnn on cellular behavior may be linked to the regulation of the expression of specific subset genes. This subset includes cell cycle-related genes such as p21(cip1/waf1), CDK4, CPR2; cell migration and invasion regulatory genes such as RhoA, CDK5, TIMP-1, MMP-7, and EMMPRIN; and MIC-1. Concordant with previous observations of Pnn-induced phenotype changes, genes coding for epithelial associated processes and cell division controls were elevated, while those coding for increased cell motility and cellular reorganizations were downregulated. We utilized p21 promoter-luciferase reporter constructs and demonstrated that a marked stimulation of p21 promoter activity in 293 cells correlated with increased Pnn expression. Taken together, these data indicate that Pnn may participate in the regulation of gene expression, thereby, positively promoting cell-cell adhesion, and negatively affecting cell migration and cell proliferation. PMID- 11494130 TI - A functional screen for genes inducing epidermal growth factor autonomy of human mammary epithelial cells confirms the role of amphiregulin. AB - To gain better understanding of the molecular alterations responsible for the aggressive growth potential of epidermal growth factor receptor (EGFR)-positive breast cancers, we utilized an expression cloning strategy to seek gene products that mediate the EGF-independent growth of human breast cancer cells. A retroviral cDNA expression library was constructed from the EGFR-positive SUM 149PT cell line, and transduced into growth factor-dependent human mammary epithelial (HME) cells. Recipient cells were functionally selected for their ability to proliferate in serum-free, EGF-free medium. Library cDNAs were recovered from EGF-independent colonies by PCR amplification or by biological rescue. Clone H55a#1 contained a library insert encoding amphiregulin. This EGFR ligand was able to confer EGF independence when transduced into HME cells. SUM 149PT and H55a#1 cells overexpressed amphiregulin transcripts, and secreted moderate EGF-like activity in conditioned media, indicating a possible autocrine loop. EGFR membrane levels and constitutive phosphorylation were consistent with this hypothesis, as well as the sensitivity of the cells to an ErbB-specific kinase inhibitor. Expression of the WT1 Wilms' tumor suppressor gene, a transcriptional activator of amphiregulin, did not parallel amphiregulin transcript levels, suggesting that another factor regulates amphiregulin in SUM 149PT. Our data confirm the importance of amphiregulin in the etiology of breast cancer. PMID- 11494131 TI - Downregulation of the type 1 insulin-like growth factor receptor in mouse melanoma cells is associated with enhanced radiosensitivity and impaired activation of Atm kinase. AB - The type 1 insulin-like growth factor receptor (IGF1R) is required for growth, tumorigenicity and protection from apoptosis. IGF1R overexpression is associated with radioresistance in breast cancer. We used antisense (AS) RNA to downregulate IGF1R expression in mouse melanoma cells. Cells expressing AS-IGF1R transcripts were more radiosensitive in vitro and in vivo than controls. Also they showed reduced radiation-induced p53 accumulation and p53 serine 18 phosphorylation, and radioresistant DNA synthesis. These changes were reminiscent of the cellular phenotype of the human genetic disorder ataxia-telangiectasia (A-T), caused by mutations in the ATM gene. Cellular Atm protein levels were lower in AS-IGF1R transfected cells than in control cells, although there was no difference in Atm expression at the transcriptional level. AS-IGF1R cells had detectable basal Atm kinase activity, but failed to induce kinase activity after irradiation. This suggests that IGF1R signalling can modulate the function of Atm, and supports the concept of targeted IGF1R downregulation as a potential treatment for malignant melanoma and other radioresistant tumours. PMID- 11494132 TI - An alternatively spliced HDM2 product increases p53 activity by inhibiting HDM2. AB - The human counterpart hdm2 of the murine double-minute 2 (mdm2) gene encodes a 90 kD protein (HDM2) that inhibits the function of the p53 tumor suppressor. Hdm2 is amplified in approximately 30% of sarcomas, leading to overproduction of HDM2 and inactivation of p53. Using immunohistochemistry to screen a panel of human tumors for HDM2 overproduction, we detected high levels of HDM2 in the cytoplasm in 25% of lung tumors as opposed to its normal localization in the nucleus. These samples contained full-length hdm2 and several alternate-splice forms of hdm2 mRNA. Sequence analysis revealed deletions in the alternate-splice forms of the p53 binding domain and absence of a nuclear localization signal. In transient transfection assays, one of the alternate-splice forms, HDM2(ALT1), bound and sequestered full-length HDM2 in the cytoplasm. In addition, the binding of HDM2(ALT1) to HDM2 inhibited the interaction of HDM2 with p53, thus enhancing p53 transcriptional activity. These data suggest the existence of another level of regulation of HDM2 which increases the activity of p53. PMID- 11494133 TI - p73 cooperates with DNA damage agents to induce apoptosis in MCF7 cells in a p53 dependent manner. AB - p73, a member of the p53 family, can induce apoptosis in cancer cells. Since p53 mediated apoptosis can be augmented by various cancer chemotherapeutic agents, it has been hypothesized that the status of the endogenous p53 gene in cancer cells is a key determinant in the outcome of cancer therapy. To determine whether p73 can sensitize cancer cells to apoptosis by DNA damage agents, several MCF7 adenocarcinoma cell lines that inducibly express p73 or p53 under a tetracycline regulated promoter were generated. We found that at relevant physiological levels, p73, but not p53, is capable of sensitizing MCF7 cells to apoptosis induced by chemotherapeutic agents. In addition, we found that p73 can cooperate with the DNA damaging agent camptothecin to activate the initiator caspase 2. Furthermore, we found that p73 can cooperate with DNA damaging agents or p53 to induce some p53 target genes and activate their promoters. In contrast, in MCF7E6 cells that ectopically express the human papillomavirus E6 oncogene and are functionally p53-null, the ability of p73 to sensitize cells to apoptosis is abrogated. Taken together, these results suggest that a functional interaction between p53 and p73 in MCF7 cells leads to enhanced induction of apoptosis. PMID- 11494134 TI - Regulation of Cbl phosphorylation by the Abl tyrosine kinase and the Nck SH2/SH3 adaptor. AB - The Cbl proto-oncogene product is tyrosine phosphorylated in response to a wide variety of stimuli. Cbl and the Abl nonreceptor tyrosine kinase both bind to SH3 domains from the SH2/SH3 adaptor Nck, and are candidate effectors for Nck function. Numerous additional SH2- and SH3-domain-mediated interactions are also possible between Cbl, Abl, and Nck. We find that these three signaling proteins associate when overexpressed in mammalian cells and can regulate each other's activity. Co-expression of wt Cbl together with c-Abl, the activity of which is normally repressed in vivo, led to extensive Abl-dependent phosphorylation of Cbl. The major proline-rich region of Cbl was required for its phosphorylation by c-Abl, but not by a constitutively activated Abl mutant, suggesting Cbl activates c-Abl by engaging its SH3 domain. Efficient phosphorylation of Cbl and its stable association with Abl required the SH2 domain of Abl, suggesting that SH2 phosphotyrosine interactions prevent dissociation of active Abl from Cbl. We also show that overexpression of Nck could repress the phosphorylation of Cbl by Abl in vivo. Studies with Nck mutants suggested that the Nck SH2 domain is responsible for inhibiting the activity of Abl toward both Cbl and Nck itself, most likely by competing with the Abl SH2 for tyrosine-phosphorylated binding sites. PMID- 11494135 TI - Mechanism of colon cancer cell apoptosis mediated by pyropheophorbide-a methylester photosensitization. AB - Pyropheophorbide-a methylester (PPME) is a second generation of photosensitizers used in photodynamic therapy (PDT). We demonstrated that PPME photosensitization triggered apoptosis of colon cancer cells as measured by using several classical parameters such as DNA laddering, PARP cleavage, caspase activation and mitochondrial release of cytochrome c. Preincubation of cells with N-acetyl cysteine (NAC) or pyrolidine dithiocarbamate (PDTC) protected against apoptosis mediated by PPME photosensitization showing that reactive oxygen species (ROS) are involved as second messengers. On the other hand, photosensitization carried out in the presence of deuterium oxide (D2O) which enhances singlet oxygen (1O2) lifetime only increases necrosis without affecting apoptosis. Since PPME was localized in the endoplasmic reticulum (ER)/Golgi system and lysosomes, other messengers than ROS were tested such as calcium, Bid, Bap31, phosphorylated Bcl-2 and caspase-12 but none was clearly identified as being involved in triggering cytochrome c release from mitochondria. On the other hand, we demonstrated that the transduction pathways leading to NF-kappaB activation and apoptosis were clearly independent although NF-kappaB was shown to counteract apoptosis mediated by PPME photosensitization. PMID- 11494136 TI - Mitochondrial cytochrome c release is caspase-dependent and does not involve mitochondrial permeability transition in didemnin B-induced apoptosis. AB - Permeability transition, and a subsequent drop in mitochondrial membrane potential (DeltaPsi(m)), have been suggested to be mechanisms by which cytochrome c is released from the mitochondria into the cytosol during apoptosis. Furthermore, a drop in DeltaPsi(m) has been suggested to be an obligate early step in the apoptotic pathway. Didemnin B, a branched cyclic peptolide described to have immunosuppressive, anti-tumour, and anti-viral properties, induces rapid apoptosis in a range of mammalian cell lines. Induction of apoptosis by didemnin B in cultured human pro-myeloid HL-60 cells is the fastest and most complete ever described with all cells being apoptotic after 3 h of treatment. By utilizing the system of didemnin B-induced apoptosis in HL-60 cells, and the potent inhibitors of mitochondrial permeability transition, cyclosporin A and bongkrekic acid, we show that permeability transition as determined by changes in DeltaPsi(m) and mitochondrial Ca2+ fluxing, is not a requirement for apoptosis or cytochrome c release. In this system, changes in mitochondrial membrane potential and cytochrome c release are shown to be dependent on caspase activation, and to occur concurrently with the release of caspase-9 from mitochondria, genomic DNA fragmentation and apoptotic body formation. PMID- 11494137 TI - The p36 isoform of BAG-1 is translated by internal ribosome entry following heat shock. AB - BAG-1 (also known as RAP46/HAP46) was originally identified as a 46 kDa protein that bound to and enhanced the anti-apoptotic properties of Bcl-2. BAG-1 exists as three major isoforms (designated p50, p46 and p36 or BAG-1L, BAG-1M and BAG-1S respectively) and one minor isoform (p29), which are translated from a common transcript. The differing amino terminus determines both the intracellular location and the repertoire of binding partners of the isoforms which play different roles in a variety of cellular processes including signal transduction, heat shock, apoptosis and transcription. Although in vitro data suggest that the four BAG-1 isoforms are translated by leaky scanning, the patterns of isoform expression in vivo, especially in transformed cells, do not support this hypothesis. We have performed in vivo analysis of the BAG-1 5' untranslated region and shown that translation initiation of the most highly expressed isoform (p36/BAG-1S) can occur by both internal ribosome entry and cap-dependent scanning. Following heat shock, when there is a downregulation of cap-dependent translation, the expression of the p36 isoform of BAG-1 is maintained by internal ribosome entry. PMID- 11494138 TI - Differential activation of TRAIL-R1 and -2 by soluble and membrane TRAIL allows selective surface antigen-directed activation of TRAIL-R2 by a soluble TRAIL derivative. AB - TNF-related apoptosis-inducing ligand (TRAIL) is a typical member of the tumor necrosis factor (TNF) ligand family that is expressed as a type II membrane protein (memTRAIL) and signals apoptosis via the death domain-containing receptors TRAIL-R1 and -2. Soluble recombinant derivatives of TRAIL (sTRAIL) are considered as novel tumors therapeutics because of their selective apoptosis inducing activity in a variety of human tumors but not in normal cells. Using antagonistic antigen-binding fragment (Fab) preparations of TRAIL-R1- and TRAIL R2-specific antibodies, we demonstrate in this study that TRAIL-R1 becomes activated by both the soluble and the membrane-bound form of the ligand, whereas TRAIL-R2 becomes only activated by memTRAIL or soluble TRAIL secondarily cross linked by antibodies. Furthermore, we show that the restricted signal capacity of sTRAIL can be readily converted into a fully signal competent memTRAIL-like molecule, i.e. a TRAIL-R2 stimulating ligand, by genetic fusion to an antibody derivative that allows antigen-dependent 'immobilization' of the fusion protein to cell surfaces. We conclude that antibody targeting-dependent activation can be used to design selective therapeutics derived of those ligands of the TNF family that are biologically inactive in their soluble form. PMID- 11494139 TI - Tissue-specific expression of SV40 in tumors associated with the Li-Fraumeni syndrome. AB - Inactivation of wild-type p53 tumor suppressor function is the primary mechanism of tumor initiation in Li-Fraumeni syndrome (LFS) individuals with germline p53 mutations. Tumors derived from LFS patients frequently retain the normal p53 allele, suggesting that alternative mechanisms in addition to gene deletion must be involved in inactivating wild-type p53 protein. DNA tumor viruses, such as SV40, target p53 for inactivation through the action of viral oncoproteins. We studied the probands from two unrelated LFS families, each of whom presented with multiple malignant neoplasms. Patient 1 developed an embryonal rhabdomyosarcoma (RMS) and a choroid plexus carcinoma (CPC), while patient 2 developed a CPC and subsequently presented with both an osteosarcoma (OS) and renal cell carcinoma (RCC). We utilized DNA sequence analysis and immunohistochemistry to determine p53 gene status in the germline and tumors, as well as evidence for SV40 T antigen oncoprotein expression. Each patient harbored a heterozygous germline p53 mutation at codons 175 and 273, respectively. In patient 1, the normal p53 gene was lost while the mutant p53 allele was reduced to homozygosity in the RMS. Both normal and mutant genes were maintained in the CPC. In patient 2, normal and mutant p53 alleles were retained in both the CPC and RCC. Both specific PCR and immunostaining detected SV40 T-antigen in both CPCs and the RCC. In addition to chromosomal alterations, epigenetic mechanisms may disrupt p53 function during tumorigenesis. In two LFS patients, we found SV40 DNA sequences and viral T antigen expression that could account for inactivation of the normal p53 protein. Inactivation of p53 or other tumor suppressors by viral proteins may contribute to tumor formation in specific tissues of genetically susceptible individuals. PMID- 11494140 TI - Detection of differentially expressed genes in human colon carcinoma cells treated with a selective COX-2 inhibitor. AB - Numerous reports suggest that use of nonsteroidal anti-inflammatory drugs (NSAIDs) decrease mortality from colorectal cancer. To better understand all of the mechanisms underlying this effect, the global pattern of gene expression in colon carcinoma cells following treatment with NS-398, a selective cyclo oxygenase-2 inhibitor was evaluated. We utilized suppression subtractive hybridization combined with differential screening to identify genes whose expression was affected following treatment. Among the subtracted cDNA fragments confirmed as differentially expressed, there were two which are known to be involved in the regulation of cell adhesion (human FAT and proto-cadherin-7). We identified two other genes whose levels were decreased and these are known to be involved in the regulation of cell proliferation (cyclin K and p-100). We identified additional genes which are involved in different signaling pathways which regulate programmed cell death (Dynamin 2, Pdcd4 and LIP.1). These results provide evidence that some of the effects of NS-398 on carcinoma cells may be due to modulation of genes which regulate programmed cell death, cell proliferation and cell-cell communication. Additional studies are underway to determine the biological function of the novel genes that were identified. PMID- 11494141 TI - Growth-suppressive effects of BPOZ and EGR2, two genes involved in the PTEN signaling pathway. AB - Defects in PTEN, a tumor suppressor, have been found in cancers arising in a variety of human tissues. To elucidate the tumor-suppressive function of this gene, we have been analysing expression profiles of cancer cells after introduction of exogenous PTEN. Those experiments identified 99 candidate genes that were transcriptionally transactivated. Among them, we report here the further analyses of eight genes, EGR2/Krox-20, BPOZ, APS, HCLS1/HS1, DUSP1/MKP1, NDRG1/Drg1/RTP, NFIL3/E4BP4, and a novel gene (PINK1, PTEN-induced putative kinase). Expression of six of them (PINK1, EGR2, HCLS1, DUSP1, BPOZ, and NFIL3) was decreased in ovarian tumors compared with corresponding normal tissues. Colony-formation assays using plasmid clones designed to express each gene indicated that EGR2 and BPOZ were able to suppress growth of cancer cells significantly; in particular, cancer-cell lines stably expressing BPOZ grew more slowly than control cells containing mock vector. Flow cytometry suggested that over-expression of BPOZ inhibited progression of the cell cycle at the G(1)/S transition. Anti-sense oligonucleotides for BPOZ or EGR2 effectively inhibited their expression, and cell growth was accelerated. Therefore both genes appear to be novel candidates as mediators of the PTEN growth-suppressive signaling pathway. PMID- 11494142 TI - Biochemical differences between SUDHL-1 and KARPAS 299 cells derived from t(2;5) positive anaplastic large cell lymphoma are responsible for the different sensitivity to the antiproliferative effect of p27(Kip1). AB - An inverse correlation between p27(Kip1) expression and proliferation has been recently established in tissues derived from human lymphomas. The nucleophosmin anaplastic lymphoma kinase (NPM-ALK)/phospholipase C-gamma (PLCgamma) complex also appears to play an important role in cell proliferation and malignant transformation of anaplastic large cell lymphoma (ALCL). In this study, we report that SUDHL-1 and KARPAS 299 ALCL-derived cell lines present different sensitivity to the antiproliferative effect of recombinant adenovirus-mediated p27(Kip1) expression or to serum-starvation in culture media. The results indicate that exogenous p27(Kip1) may interact with the NPM-ALK/PLCgamma pathway in SUDHL-1 but not in KARPAS 299 cells. This interaction correlates with changes in cell cycle and cell morphology observed mainly in SUDHL-1 cells. The percentage of SUDHL-1 cells in S phase declines, whereas it is almost unchanged in KARPAS 299 cells as compared to the controls after 96 h of infection with the recombinant adenovirus. Furthermore KARPAS 299 cells are resistant to serum-starvation due to deficient p27(Kip1)-upregulation and G1 arrest, whereas SUDHL-1 cells respond with increased G1 phase and p27(Kip1)-upregulation after 48 h of serum-starvation. Both cell lines express appropriate variation of levels of cyclins E and A, and Rb-phosphorylation as expected by growing them in culture media with different FBS content. Although both cell lines express cyclin D2, SUDHL-1 cells only present high level of cyclin D3. Moreover SUDHL-1 cells express high level of PTEN and the PKB/Akt pathway is constitutively activated in both cell lines. Lastly SUDHL-1 cells show higher levels of phosphotyrosine-containing proteins that is correlated with a higher NPM-ALK-associated autophosphorylation activity compared to KARPAS 299 cells. Our study clearly identifies some of the biochemical differences that may explain the difference in sensitivity to antiproliferative stimuli shown by two cell lines derived from the same type of lymphoma. PMID- 11494143 TI - Role of Bax in apoptosis of IL-3-dependent cells. AB - IL-3 removal was reported to induce membrane association of the apoptotic effector Bax. This report demonstrates that IL-3-dependent cells from Bax-null mice failed to activate caspases after IL-3 removal and survived in an 10-fold lower concentration of IL-3. As IL-3 removal also down-regulates expression of Bcl-X, we examined the relationship between Bcl-X decrease and Bax membrane association. IL-3 removal from BAF-3 cells, followed by sorting caspase-active and caspase-inactive populations, showed that both expressed similar levels of Bcl-X. Inhibition of IL-3 signalling via PI-3 kinase and MEK1/2 resulted in cells with minimal Bcl-X, which remained viable with soluble Bax. However BAF-3-derived cells, which maintained Bcl-X expression without IL-3, also remained viable with soluble Bax on IL-3 removal. Therefore a decrease in Bcl-X is necessary, though not sufficient, for Bax membrane association on IL-3 removal. In contrast, treatment of BAF-3 cells with hydroxyurea induced apoptosis in the absence of a Bcl-X decrease. Furthermore, IL-3-dependent cells from Bax-null mice activated caspases after hydroxyurea treatment and show the same sensitivity to a variety of cytotoxic drugs. Thus, apoptosis after IL-3 removal requires a decrease in Bcl X and Bax membrane association, whereas that induced by cytotoxic drugs does not. PMID- 11494144 TI - Genome-wide expression changes induced by HTLV-1 Tax: evidence for MLK-3 mixed lineage kinase involvement in Tax-mediated NF-kappaB activation. AB - The Tax protein of human T-lymphotropic virus type 1 (HTLV-1), an oncoprotein that transactivates viral and cellular genes, plays a key role in HTLV-1 replication and pathogenesis. We used cDNA microarrays to examine Tax-mediated transcriptional changes in the human Jurkat T-cell lines JPX-9 and JPX-M which express Tax and Tax-mutant protein, respectively, under the control of an inducible promoter. Approximately 300 of the over 2000 genes examined were differentially expressed in the presence of Tax. These genes were grouped according to their function and are discussed in the context of existing findings in the literature. There was strong agreement between our results and genes previously reported as being Tax-responsive. Genes that were differentially expressed in the presence of Tax included those related to apoptosis, the cell cycle and DNA repair, signaling factors, immune modulators, cytokines and growth factors, and adhesion molecules. Functionally, we provide evidence that one of these genes, the mixed-lineage kinase MLK-3, is involved in Tax-mediated NF-kappa B signaling. Our current results provide additional insights into Tax-mediated signaling. PMID- 11494145 TI - Induction of immune responses and safety profiles in rhesus macaques immunized with a DNA vaccine expressing human prostate specific antigen. AB - Prostate specific antigen (PSA) is a widely used marker for prostate cancer, which is secreted by normal prostate cells at low levels, but is produced more substantially by cancer cells. We have previously reported on the use of a DNA vaccine construct that encodes for human PSA gene to elicit host immune responses against cells producing PSA. DNA immunization strategy delivers DNA constructs encoding for a specific immunogen into the host, who becomes the in vivo protein source for the production of antigen. This antigen then is the focus of the resulting immune response. In this study, we examine the induction of immune responses and safety profiles in rhesus macaques immunized with DNA-based PSA vaccine. We observed induction of PSA-specific humoral response as well as positive PSA-specific lymphoproliferative (LPA) response in the vaccinated macaques. We also observed that the stimulated T cells from the PSA-immunized rhesus macaques produced higher levels of Th1 type cytokine IFN-gamma than the control vector immunized animals. On the other hand, DNA immunization did not result in any adverse effects in the immunized macaques, as indicated by complete blood counts, leukocyte differentials and hepatic and renal chemistries. The macaques appeared healthy, without any physical signs of toxicity throughout the observation period. In addition, we did not observe any adverse effect on the vaccination site. The apparent safety and immunogenecity of DNA immunization in this study suggest that further evaluation of this vaccination strategy is warranted. PMID- 11494146 TI - BAD/BCL-[X(L)] heterodimerization leads to bypass of G0/G1 arrest. AB - The pro-apoptotic molecule BAD binds BCL-[X(L)] or BCL2 and inactivates their survival function. In addition to their anti-apoptotic function, BCL2 and BCL [X(L)] also delay cell cycle entry from quiescence. We found that the BH3-only molecule BAD also exerted a cell cycle effect. BAD expression resulted in failure to cell cycle block in growth arrest conditions. In low serum and in confluence, fibroblasts constitutively or inducibly expressing BAD persisted in S phase, continued to incorporate BrdU, and exhibited sustained cyclin E/cdk2 activity. Mutation analysis indicated that the cell cycle effect of BAD was not dependent on its phosphorylation status or subcellular localization, but strictly co segregated with BCL-[X(L)] binding. bclx(-/-) MEFs expressing BAD and bad(-/-) MEFs both arrested in G0/G1 in low serum similar to wild-type controls, suggesting that the ability to overcome the G0/G1 checkpoint resulted from the presence of BAD/BCL-x(L) heterodimers, rather than the absence of BCL-[X(L)] or BAD. These data provide evidence that in addition to regulating apoptosis, the BAD/BCL-[X(L)] heterodimer has a novel cell cycle function. PMID- 11494147 TI - The role of tumor necrosis factor alpha in the pathophysiology of human multiple myeloma: therapeutic applications. AB - In this study we demonstrate that tumor necrosis factor alpha (TNFalpha) triggers only modest proliferation, as well as p44/p42 mitogen-activated protein kinase (MAPK) and NF-kappaB activation, in MM.1S multiple myeloma (MM) cells. TNFalpha also activates NF-kappaB and markedly upregulates (fivefold) secretion of interleukin-6 (IL-6), a myeloma growth and survival factor, in bone marrow stromal cells (BMSCs). TNFalpha in both a dose and time dependent fashion induced expression of CD11a (LFA-1), CD54 (intercellular adhesion molecule-1, ICAM-1), CD106 (vascular cell adhesion molecule-1, VCAM-1), CD49d (very late activating antigen-4, VLA-4), and/or MUC-1 on MM cell lines; as well as CD106 (VCAM-1) and CD54 (ICAM-1) expression on BMSCs. This resulted in increased (2-4-fold) per cent specific binding of MM cells to BMSCs, with related IL-6 secretion. Importantly, the proteasome inhibitor PS-341 abrogated TNFalpha-induced NF-kappaB activation, induction of ICAM-1 or VCAM-1, and increased adhesion of MM cells to BMSCs. Agents which act to inhibit TNFalpha may therefore abrogate the paracrine growth and survival advantage conferred by MM cell adhesion in the BM microenvironment. PMID- 11494148 TI - STAT3 activation is required for Asp(816) mutant c-Kit induced tumorigenicity. AB - Activating mutations of c-kit at codon 816 (Asp(816)) have been identified in variety of malignancies, including acute myeloid leukemia (AML), mastocytosis and germ cell tumors. The mutant c-Kit receptor confers cytokine independence and induces tumorigenicity. However, the molecular mechanisms, particularly the changes in the signal transduction pathways, responsible for these biological effects induced by mutant c-Kit are largely undefined. Using the human embryonic kidney cell line, 293, we show in the current report that constitutive activation of STAT3 and STAT1 is associated with D816H mutant c-Kit. Transfection of dominant negative STAT3, but not STAT1 inhibits mutant c-Kit mediated anchorage independent growth in vitro and tumor formation in vivo. Expression of constitutively activated STAT3 restores the mutant c-Kit receptor's transforming ability in 293 cells. These results demonstrate that activation of STAT3 by Asp(816) mutant c-Kit is required for the anchorage-independent growth and tumorigenicity induced by Asp(816) mutant c-Kit. PMID- 11494149 TI - The expression of HMGA genes is regulated by their 3'UTR. AB - Many benign mesenchymal tumors are characterized by chromosomal abnormalities of the regions 12q15 or 6p21.3 leading to aberrant expression of either HMGA2 (formerly HMGIC) or HMGA1 (formerly HMGIY). The proteins of both genes belong to the HMGA (formerly HMGI(Y)) family of architectural transcription factors. As a rule, aberrant HMGA transcripts found in a variety of benign tumors have intact coding regions at least for the DNA binding domains with a truncation of their 3' untranslated regions. Adding this to the finding that an altered HMGA protein level is not always correlated with an increased amount of corresponding mRNA indicates a posttranscriptional expression control mediated by regulatory elements within the 3'UTR. To check if HMGA expression is under control of such elements we performed luciferase assays with several HMGA2 and HMGA1 3'UTRs of different length. Experiments showed that an up to 12-fold increase in luciferase activity is obtained by the truncation of the 3'UTRs suggesting that the expression of HMGA2 and HMGA1 is controlled by negatively acting regulatory elements within their 3'UTR. Chromosomal aberrations affecting the HMGA genes may therefore influence their expression by an altered stability of the truncated transcripts as a result of the cytogenetic aberrations. PMID- 11494150 TI - Cdc25A phosphatase suppresses apoptosis induced by serum deprivation. AB - The phosphatase Cdc25A was shown to be a target of the transcription factor c Myc. Myc-induced apoptosis appeared dependent on Cdc25A expression and Cdc25A over-expression could substitute for Myc-triggered apoptosis. These findings suggested that an important downstream component of Myc-mediated apoptosis was identified. However and in contrast, we recently reported that during TNFalpha induced apoptosis, which required c-Myc function, Cdc25A was down-regulated in a human carcinoma cell line. We now provide evidence that Cdc25A rendered the non transformed rat embryonic cell line 423 refractory to apoptosis, which was induced by serum deprivation and in absence of detectable c-myc levels. The survival promoting activity of cdc25A was abolished upon infection of cells with a full-length cdc25A antisense construct. To identify the signaling proteins mediating the survival function of the phosphatase, cdc25A- and akt- over expressing pooled clones were exposed to selected chemicals, which inhibit or activate key proteins in signaling pathways. Inhibition of apoptosis by SU4984, NF023 and Rapamycin placed Cdc25A and Akt function downstream of FGF.R, PDGF.R, and compensated G-protein- and PP2A- activity. Interestingly, upon treatment with LY-294002, cdc25A- and akt- over-expressing clones exhibited similar apoptotic patterns as control cells, which indicates that neither Akt- nor Cdc25A-mediated survival functions are dependent on PI.3 kinase activity in rat 423 cells. In cdc25A-overexpressing cells increased levels of serine 473 phosphorylated Akt were found, which co-precipitated with Cdc25A and Raf1. Since activation of proteins requires dephosphorylation of particular residues in addition to site specific phosphorylation, the anti-apoptotic effect of Cdc25A might derive from its participation in a multimeric protein complex with phosphoAkt and Raf1, two prominent components of survival pathways. PMID- 11494151 TI - Adhesion-regulated G1 cell cycle arrest in epithelial cells requires the downregulation of c-Myc. AB - Adhesion to the extracellular matrix is required for the expression and activation of the cyclin-cyclin-dependent kinase (CDK) complexes, and for G1 phase progression of non-transformed cells. However, in non-adherent cells no molecular mechanism has yet been proposed for the cell adhesion-dependent up regulation of the p27 cyclin-dependent kinase inhibitor (CKI), and the associated inhibition of cyclin E-CDK2. We now show that in epithelial cells the expression of c-Myc is tightly regulated by cell-substrate adhesion. When deprived of adhesion, two independently derived mammary epithelial cell lines, 184A1N4 and MCF-10A, rapidly decrease their level of c-Myc mRNA and protein. This decrease in levels of c-Myc correlates with G1 phase arrest, as indicated by hypophosphorylation of pRb and inhibition of the activity of the cyclin E-CDK2 complex. In 184A1N4 cells, cell-substrate adhesion is required for the suppression of p27, and induction of cyclin E, E2F-1, but not cyclins D1 and D3. Enforced expression of c-Myc in non-adherent 184A1N4 and MCF-10A cells reverses the adhesion-dependent inhibition of cell cycle progression. Restoration of c-Myc in non-adherent cells induces the expression of E2F-1, and hyperphosphorylation of pRb in response to EGF treatment. In addition, expression of c-Myc results in the anchorage-independent activation of the CDK2 complex, the associated upregulation of cyclin E, and the destabilization and degradation of p27 by the ubiquitin-proteasome pathway. Our study thus suggests that c-Myc is the link between cell adhesion and the regulation of p27 and cyclin E-CDK2. Furthermore, we describe a role for c-Myc in adhesion-mediated regulation of E2F-1. PMID- 11494152 TI - Expression profile of nine novel genes differentially expressed in hepatitis B virus-associated hepatocellular carcinomas. AB - Chronic hepatitis B virus (HBV) infection is known to be one of the major causes in the development of hepatocellular carcinoma (HCC), although the biomolecular mechanism(s) involved remain unclear. To identify the cellular gene(s) involved in HBV-associated hepatocarcinogenesis, we used the mRNA differential display method and examined three paired tumor and nontumor tissues, all of which had chromosomally integrated HBV-DNA through chronic infection. Using 240 different combinations of three one-base anchored oligo-dT primers and 80 arbitrary 13 mers, genes decreased or increased in expression more than twofold between each tumor tissue and its paired nontumor tissue were identified. Twenty-nine known genes and four novel genes were differentially over-expressed in the HCC tumor tissues. In contrast, 27 known genes and five novel genes were under-expressed in those tumor tissues. The nucleotide sequences of the nine novel gene fragments were determined and their expression patterns were examined in 40 HCC samples. HA61T2, PT18, HG63T1, and HG57T1 were preferentially over-expressed in 32 cases (80%, P<0.001), 24 cases (60%), 23 cases (57.5%) and 22 cases (55%) of the 40 tumor tissues, respectively. There was an increased frequency of HG57T1 over expression in HCC patients with HBV-positive serology and low serum alpha-feto protein (AFP) levels (P<0.05). DNT10, PT8, PT19, ENT25 and HA6T4 were under expressed in 26 cases (65%), 23 cases (57.5%), 21 cases (53%), 20 cases (50%) and 18 cases (45%) of the 40 tumor samples, respectively. There was a strong correlation of DNT10 under-expression with high serum AFP level in HCC patients, irrespective of HBV serology (P<0.01). HA6T4 was preferentially under-expressed in HCC tumors in patients with HBV-positive serology and high serum AFP levels (P<0.05). Thus, the functional analyses of the known and novel genes identified in this study should prove valuable to further understand the mechanism(s) of hepatocarcinogenesis. PMID- 11494153 TI - Hdmx and Mdm2 can repress transcription activation by p53 but not by p63. AB - The p53 protein is involved in cell cycle arrest and apoptosis. To ensure that cells under non-stressed conditions are able to grow, p53 sets up a negative feedback loop by inducing Mdm2. Mdm2 is able to both inhibit the transcriptional regulation by p53 and to degrade it, thus maintaining p53 inactive until it is required. The Mdm2 related protein, Hdmx, has also been shown to inhibit the transcriptional activation of p53 but is unable to degrade it. A few years ago, the p53 family member, p63 was identified. Like p53, p63 is able to induce p53 target genes and it was shown to be able to cause cell cycle arrest and apoptosis. In this study we report that, despite the similarities between p53 and p63, neither Hdmx nor Mdm2 are able to interact with p63, to repress p63-induced transcription or to affect its half-life. PMID- 11494154 TI - A rapid bioinformatic method identifies novel genes with direct clinical relevance to colon cancer. AB - Identifying genes whose differential expression affect the survival of patients after primary tumor surgery is a major aim of clinical cancer research. To address this issue we combined rapid bioinformatic search algorithms with quantitative RT-PCR in a panel of clearly defined cases of colorectal carcinomas with detailed patient histories. Search algorithms were written that identified Expressed Sequence Tags (ESTs) from the Unigene EST collection of putative open reading frames (ORFs). Expression ratios of healthy to cancerous tissue of each Unigene ORF were calculated. The first 35 candidates arising from bioinformatic searches were examined for mRNA expression in a panel of 20 well documented cases of colon cancer. Four of these 35 genes showed significant correlations with histopathological parameters. Therefore, their expression was further analysed by quantitative RT-PCR in a larger patient cohort. Kaplan-Meier/log rank statistical tests of up to 49 patients in three of the four genes demonstrated significant association of gene expression with poor survival. All four genes demonstrated a strong association with metastatic tumor progression. Expression of the genes was localized to epithelial cells by in-situ hybridization. PMID- 11494155 TI - Analysis of genetic and epigenetic processes that influence p14ARF expression in breast cancer. AB - The INK4a/ARF locus encodes two unrelated cell cycle-regulatory proteins that both function in tumor suppression, p16INK4a and p14ARF. In human tumors including breast cancer, alterations affecting selectively p14ARF have been poorly analysed. We have performed a comprehensive analysis of the inactivation mechanisms (mutation, homozygous and hemizygous deletion, and promoter hypermethylation) in a large series of 100 primary breast carcinomas. RT-PCR showed expression variable of the p14ARF transcript, with 17% demonstrating overexpression and 26% demonstrating decreased expression. No detectable alterations were observed in the majority of cases with overexpressed p14ARF mRNA, but 77% of tumors with decreased expression presented at least one of these genetic/epigenetic alterations. Nevertheless, a statistically significant correlation was observed between decreased p14ARF expression and several poor prognostic parameters. PMID- 11494156 TI - Cleavage of Bcl-2 in oxidant- and cisplatin-induced apoptosis of human melanoma cells. AB - Although the anti-apoptotic effect of Bcl-2 is well established, the role of Bcl 2 in tumour response to therapy and drug resistance is still unclear. The post translational modifications of Bcl-2 are likely involved in the control of the apoptotic pathway. In the present study we have investigated the role of Bcl-2 in cellular response to oxidative stress (hydrogen peroxide) and cisplatin using a clone of human metastatic melanoma, which, in spite of Bcl-2 (over)expression, exhibited a moderate chemosensitivity. With both treatments melanoma cells died through an apoptotic process, associated with detachment of cells from the monolayer. In the floating apoptotic cells generated by either hydrogen peroxide or cisplatin, along with morphological and biochemical features of apoptosis, we detected a significant Bcl-2 cleavage, yielding the Bax-like fragment of 23 kDa. Preincubation of cells with the caspase-3/-7 inhibitor DEVD-CHO completely suppressed Bcl-2 cleavage, thus confirming that such a specific proteolysis requires activation of caspase-3/-7. The oxidant- and cisplatin-induced processing of Bcl-2 documented in the present study may represent a regulatory mechanism to circumvent the survival function of Bcl-2 upon apoptosis triggering and to enhance apoptotic response. Since the Bcl-2 cleavage should be regarded as a pro-apoptotic event, Bcl-2 expression is expected to increase susceptibility to apoptosis. Thus, such a pathway could be exploited to improve the efficacy of cytotoxic therapy of melanomas expressing Bcl-2. PMID- 11494157 TI - A somatic mutation in the 5'UTR of BRCA1 gene in sporadic breast cancer causes down-modulation of translation efficiency. AB - Mutations in the 5' UTR which cause increment/decrement of translation efficiency have been recently described as a novel molecular mechanism of disease. Alterations in the consensus sequence for the translation initiation may promote context-dependent leaky scanning of ribosomes and/or initiation from a downstream AUG codon. Initiation of translation from a downstream in-frame AUG codon in BRCA1 gene was recently identified in normal cells and possibly in breast cancer. Here we present further insight into BRCA1 translational pathophysiology investigating the role of the canonical structure of the initiation consensus sequence of BRCA1. We have analysed the effect of a somatic point mutation (117 G>C) in position -3 with respect to the AUG of the BRCA1 gene, identified in a highly aggressive sporadic breast cancer. We constructed chimeric genes encoding the luciferase reporter sequence downstream of the wild type or the mutated BRCA1 5'UTR. These transcripts were tested for their activity in in vitro and in vivo systems. In in vitro transcription/translation assays the estimated translation efficiency of the construct with the mutated BRCA1 5'UTR was 30-50% lower than that with the wild type BRCA1 5'UTR. The same chimeric genes were analysed for their expression in vivo by transient transfection in human cells. While the two constructs were equally transcribed, the plasmid carrying the mutated sequence produced 70% less luciferase activity compared to the wild type sequence. Finally, to obtain a direct evaluation on translational efficiency in vivo, we analysed mRNA translation on translationally active and non-active ribosomes separated from transfected cells. Mutant mRNA was partially localized in subpolysomal particles analytically confirming a polysome recruitment defect. Thus, characterization of BRCA1 5'UTR and translation efficiency seems to provide new insight into BRCA1 role in breast and ovarian cancer pathogenesis. PMID- 11494158 TI - Cytomegalovirus infection in transplant recipients resolves when circulating gammadelta T lymphocytes expand, suggesting a protective antiviral role. AB - gammadelta T cells undergo massive expansion in the peripheral blood of renal transplant recipients who are infected with cytomegalovirus (CMV). In a 3-year prospective study, the relationship between the evolution of CMV infection and the kinetics of gammadelta T cell amplification was followed for 10 months after transplantation. Patients with late gammadelta T cell expansion (>/=45 days) had significantly longer (P<.0001) and higher (P<.0003) pp65 antigenemia and more symptomatic CMV disease than did patients with early expansion. Analysis of data for each patient showed that gammadelta T cell expansion is concomitant with the resolution of CMV infection and disease, regardless of the CMV serologic status of donor and recipient before transplantation. These observations point to gammadelta T cell percentage determination as a new, rapid, and reliable prognosis factor to predict the resolution of CMV infection and strongly suggest that gammadelta T cells play a protective role against CMV infection. PMID- 11494159 TI - Ferrets as a transmission model for influenza: sequence changes in HA1 of type A (H3N2) virus. AB - Ferrets were used as an animal model to study whether controlled transmission of type A influenza is similar to human transmission when sequence changes in HA1 are used as the outcome. Ferrets were infected initially with A/Sydney/5/97 (H3N2) or A/LA/1/87 (H3N2) intranasally, and transmission chains were established by housing infected ferrets with noninfected ferrets with no influenza antibody titer against the infecting virus. Ferrets infected with A/Sydney were seronegative for A/Sydney and A/LA; ferrets infected with A/LA were seronegative for A/LA but had hemagglutination inhibition titers against A/Sydney. Titers of naturally transmitted influenza were higher than those after direct intranasal infection, but lymphocyte counts from nasal washes diminished with transmission. Ferrets infected with A/LA had 2 amino acid differences in HA1 after transmission through 5 ferret cohorts, but those infected with A/Sydney had none. The results show the value of the ferret model. A/LA resembled the transmission of influenza in humans when under antibody pressure. PMID- 11494160 TI - Human papillomavirus infection and associated cervical disease in human immunodeficiency virus-infected women: effect of highly active antiretroviral therapy. AB - To determine the effect of highly active antiretroviral therapy (HAART) on high risk human papillomavirus (HR-HPV) infections and related cervical lesions, the virologic and cytologic markers of HPV infection were prospectively studied in 163 human immunodeficiency virus (HIV)-infected women, including 27 untreated, 62 treated with reverse transcriptase inhibitors, and 74 treated with HAART. A high prevalence of both infections with HR-HPV types (68%) and squamous intraepithelial lesions (SILs; low grade, 20.2%; high grade, 6.2%) was observed. The risks of infection and disease were inversely correlated with CD4 cell counts (P=.015 and P=.022, respectively). During the observation period (mean, 15.4 months; range, 6-24 months), CD4 cell counts increased significantly only in subjects receiving HAART (P<.001). Persistence of HR-HPV infection and progression of SILs were comparable in the 3 groups. These results indicate that, even in the era of HAART, HIV-infected women should be monitored carefully for the emergence of high-grade SILs and cervical cancer. PMID- 11494161 TI - Comparison of the frequency of interleukin (IL)-2-, interferon-gamma-, and IL-4 producing T cells in 2 diseases, human immunodeficiency virus types 1 and 2, with distinct clinical outcomes. AB - Human immunodeficiency virus (HIV) type 2 infection is associated with a better clinical outcome, slower rates of CD4 T cell decline, and lower viremia than is HIV-1. This study compares HIV-1 and HIV-2 in regard to the percentages of interleukin (IL)-2-, interferon (IFN)-gamma-, and IL-4-producing cells at the single-cell level, as determined by flow cytometry. At a given degree of CD4 T cell depletion, the frequency of T cells able to produce IL-2 is better preserved in HIV-2 than in HIV-1 infection, particularly within the CD4 T cell subset. As described for HIV-1 immunodeficiency, HIV-2-positive patients exhibit a marked expansion of terminally differentiated effector CD8 T cells (CD28(-)CD27(-)IFN gamma(+)). However, the proportion of CD8 T cells able to simultaneously produce IL-2 and IFN-gamma is higher in HIV-2 disease. Considering the central role of IL 2 as a lymphocyte proliferative and survival factor, these findings provide a possible immunologic basis for the distinct course of HIV-2 immunodeficiency. PMID- 11494162 TI - Interleukin-18 inhibits human immunodeficiency virus type 1 production in peripheral blood mononuclear cells. AB - Interleukin (IL)-18 is an interferon (IFN)-gamma-inducing factor and contributes to the Th1 immune response. IL-18 added after infection of peripheral blood mononuclear cells (PBMC) with monocyte-tropic human immunodeficiency virus type 1 (HIV-1) inhibited p24 antigen production by a maximum of 72%. IFN-gamma levels in these cultures were increased, and a significant inverse relationship between HIV 1 production and IFN-gamma levels was observed. A neutralizing anti-IFN-gamma antibody reversed the IL-18 inhibitory effect. Preincubation of PBMC with IL-18 before infection inhibited p24 without additional IL-18 (64%). However, compared with the degree of IL-18 inhibition observed after a 4-day culture, no additional IL-18 inhibitory effect was observed during days 5-13. IL-18 also reduced cell surface expression of the HIV-1 receptor CD4. These results demonstrate that IL 18 inhibited HIV-1 production in PBMC through intermediate IFN-gamma. Furthermore, inhibition was present during the early stages of viral infection and was associated with reduced HIV-1 receptor expression. PMID- 11494163 TI - Natural development of antibodies to pneumococcal capsular polysaccharides depends on the serotype: association with pneumococcal carriage and acute otitis media in young children. AB - To study the natural development of antibodies to pneumococcal capsular polysaccharides of types 1, 6B, 11A, 14, 19F, and 23F and its association with pneumococcal carriage and acute otitis media (AOM), 329 children were followed-up prospectively during their first 2 years of life. Nasopharyngeal carriage was determined by cultures of nasopharyngeal swab samples, and etiology of AOM was determined by cultures of middle ear fluid. Antibodies were measured in serum samples collected at 6, 12, 18, and 24 months by EIA. Antibodies increased modestly but significantly with age. Contact with serotypes 11A and 14 was associated with increased antibody concentration as early as age 6 months. Children with contact with serotypes 6B, 19F, and 23F had antibody levels similar to those in children without contact. Antibodies increased modestly, even in children without known contact with Streptococcus pneumoniae and in children with contact with heterologous serotypes. Antibody concentrations were equal after carriage or AOM. PMID- 11494164 TI - Do antibodies to pneumococcal surface adhesin a prevent pneumococcal involvement in acute otitis media? AB - Antibodies to the pneumococcal (Pnc) surface protein PsaA are immunogenic and protective in experimental animal models, but their role in protection from Pnc disease in humans is not known. In the present study, the ability of antibodies to PsaA to prevent the progression of Pnc carriage to Pnc acute otitis media (Pnc AOM) was evaluated. Antibodies to PsaA were measured in acute-phase serum samples of children with AOM and with Streptococcus pneumoniae cultured from the nasopharynx. The risk of Pnc AOM was evaluated by a logistic regression model with anti-PsaA concentration as the predictive variable. Higher concentrations of antibodies to PsaA were associated with lower risk of the Pnc nasopharyngeal carriage progression to Pnc AOM. This was true in children 9-24 months old (odds ratio [OR], 0.49; 95% confidence interval [CI], 0.31-0.78) but not in children <9 months old (OR, 0.81; 95% CI, 0.48-1.35). PMID- 11494165 TI - Platelet microbicidal activity is an important defense factor against viridans streptococcal endocarditis. AB - To study the role of platelet microbicidal activity in host defense against infective endocarditis (IE) due to viridans streptococci (VS), the susceptibility to platelet releasate of blood and oral VS isolates from patients with and without IE was compared. The influence of neutralization of platelet microbicidal activity was studied in 2 experimental IE models. Resistance to platelet releasate was more prevalent among VS from blood of patients with IE than from blood of bacteremic patients without IE and among oral VS isolates. Serum from rabbits vaccinated with human platelet sonicate supernatants neutralized human and rabbit platelet-released microbicidal activity and had antibodies recognizing microbicidal proteins thrombocidin-1 and -2 and other human platelet proteins. In the 2 rabbit IE models, vaccination increased the susceptibility to experimental IE due to platelet releasate-susceptible VS. Thus, platelet-released microbicidal activity is an important host defense factor against IE due to VS. PMID- 11494166 TI - Impaired antifungal effector activity but not inflammatory cell recruitment in interleukin-6-deficient mice with invasive pulmonary aspergillosis. AB - A murine model of infection, in which immunocompetent or immunosuppressed interleukin-6-deficient (IL-6(-/-)) mice were infected intranasally with Aspergillus fumigatus conidia and were monitored for parameters of fungal colonization and innate and adaptive immunity, was used to assess the role of IL 6 in invasive pulmonary aspergillosis (IPA). The results indicate that IL-6(-/-) mice were more susceptible than wild-type mice to IPA. Susceptibility was associated with increased inflammatory pathology, decreased antifungal effector functions of phagocytes, and impaired development of protective type 1 responses. Exposure to exogenous IL-6 restored antifungal effector activity. PMID- 11494167 TI - Acquisition and decay of antibodies to pregnancy-associated variant antigens on the surface of Plasmodium falciparum-infected erythrocytes that protect against placental parasitemia. AB - Otherwise clinically immune women in areas endemic for malaria are highly susceptible to Plasmodium falciparum malaria during their first pregnancy. Pregnancy-associated malaria (PAM) is characterized by placental accumulation of infected erythrocytes that adhere to chondroitin sulfate A (CSA). Susceptibility to PAM decreases with increasing parity, apparently due to acquisition of antibodies directed against the variant surface antigens (VSAs) that mediate the adhesion to CSA (VSA(CSA)). This study found that levels of VSA(CSA)-specific antibodies depend on endemicity, that anti-VSA(CSA) IgG is acquired during gestation week 20, and that plasma levels of the antibodies decline during the postpartum period. There is evidence that VSA(CSA)-specific antibodies are linked to placental infection and that high antibody levels contribute to the control of placental infection by inhibiting parasite adhesion to CSA. Data suggest that VSA(CSA) is a target for vaccination against PAM. PMID- 11494168 TI - The influence of placental malaria infection and maternal hypergammaglobulinemia on transplacental transfer of antibodies and IgG subclasses in a rural West African population. AB - Two hundred thirteen mother-baby pairs in The Gambia were studied to determine the influence of placental malaria infection and maternal hypergammaglobulinemia on transplacental antibody transfer. Antibody transfer for herpes simplex virus 1 (HSV-1), respiratory syncytial virus (RSV), and varicella-zoster virus (VZV) was significantly reduced by placental malaria infection by 69%, 58%, and 55%, respectively. Maternal hypergammaglobulinemia was associated with a significant reduction in antibody transfer for HSV-1, RSV, VZV, and pneumococcus by 89%, 90%, 91%, and 88%, respectively. In addition, placental malaria infection was associated with a significant reduction in transfer of IgG1, IgG2, and IgG4 (P<.01, P=.01, and P=.03, respectively) but not of IgG3 (P=.59). Maternal hypergammaglobulinemia significantly impaired the transfer of IgG1 and IgG2 (P=.01) but not of IgG3 or IgG4 (P=.62 and P=.59, respectively). Placental malaria infection and maternal hypergammaglobulinemia were associated with reduction in the transplacental transfer of these specific antibodies, IgG1, and IgG2 in this Gambian population. PMID- 11494169 TI - A functional polymorphism of toll-like receptor 4 is not associated with likelihood or severity of meningococcal disease. AB - Human Toll-like receptor 4 (TLR4) transduces proinflammatory cytokine release by human cells in response to lipopolysaccharide (LPS). This study tested the hypothesis that, if TLR4 is rate limiting for a successful response to bacterial LPS in humans, a human gene polymorphism that results in the amino acid substitution Asp299Gly and causes reduced expression and function of TLR4 should influence susceptibility to or severity of natural gram-negative infection. The allele frequency of the Asp299Gly polymorphism was 5.9% among 879 blood donors, 6.5% among 1047 patients with microbiologically proven meningococcal disease, and 4.1% among 86 patients who died of meningococcal disease. No significant differences were observed, including those analyzed after stratification of the infected population by age and by meningococcal serogroup. Therefore, this functional TLR4 polymorphism does not influence susceptibility to or severity of meningococcal disease. PMID- 11494170 TI - Local production of anti-vibrio cholerae mucosal antibody in reproductive tract tissues after cholera. AB - To investigate whether intestinal presentation of an antigen by Vibrio cholerae, a noninvasive organism, could induce an anatomically distant mucosal immune response in reproductive tract tissues, the endocervical immune responses of women in Bangladesh were evaluated after cholera. Endocervical secretions were analyzed for secretory IgA (sIgA) antibody against the B subunit of cholera toxin (CtxB) in 9 women with cholera and 8 women with diarrhea caused by neither V. cholerae nor heat labile enterotoxin-producing Escherichia coli. Women infected with V. cholerae developed significant sIgA anti-CtxB responses in endocervical samples (P< or =.02). Antibody subtype analysis of endocervical IgA was consistent with local mucosal production (P< or =.001). Women with cholera did not develop sIgA anti-CtxB responses in serum. The ability to generate specific mucosal immune responses in reproductive tract tissues after intestinal presentation of antigen could facilitate development of vaccines effective against reproductive tract pathogens. PMID- 11494171 TI - Azithromycin chemoprophylaxis. PMID- 11494173 TI - Assay validation is needed before presumption of Escherichia coli O157:H7 infection in persons with hemolytic uremic syndrome. PMID- 11494175 TI - Is the prevalence of hepatitis C virus (HCV) RNA in anti-HCV-positive injection drug users positively correlated with age? PMID- 11494176 TI - Meningococcal serogroup a avidity indices as a surrogate marker of priming for the induction of immunologic memory after vaccination with a meningococcal A/C conjugate vaccine in infants in the United Kingdom. PMID- 11494177 TI - Prolactin and NK cells in maternal malaria. PMID- 11494178 TI - Medicare health maintenance organizations and traditional coverage: perceptions of health care among beneficiaries with disabilities. AB - OBJECTIVE: To determine whether perceptions of access to, affordability of, and quality of health care services differ for Medicare beneficiaries with disabilities in health maintenance organizations (HMOs) and traditional Medicare coverage. DESIGN: Bivariate and multivariate analyses were conducted to determine the relationship between Medicare coverage type and perceptions of health care access, affordability, and quality. SETTING: Noninstitutionalized Medicare beneficiaries across the United States. PARTICIPANTS: A random sample of 6116 beneficiaries who qualify for Medicare as working-age disabled (n = 2250), or who qualify as elderly and have at least 1 instrumental activities of daily living limitation (n = 3866). This subsample of the nationally representative Medicare Current Beneficiary Survey (MCBS) represents 11,627,107 beneficiaries with disabilities. INTERVENTION: Questions about perception of access to primary care, affordability of care, and quality of care. Data derived from the 1994 MCBS Access to Care File. MAIN OUTCOME MEASURES: DEPENDENT VARIABLES: perceptions of access to primary care, affordability, and satisfaction with quality of care. INDEPENDENT VARIABLES: indicators of managed care status and health status level, severity of disability, Medicare qualification, age, and gender. RESULTS: Beneficiaries with disabilities in HMOs perceive better access to primary care services, and greater affordability of health services than those with traditional Medicare coverage. Beneficiaries in poor health or with the most severe disabilities were most likely to perceive access and cost difficulties, regardless of coverage type. CONCLUSION: Medicare managed care appears generally to be meeting 2 of its goals-better access to primary care and more affordable care-though these advantages are not being shared by those with poor health status and/or severe disabilities. PMID- 11494179 TI - The use of proxies in community integration research. AB - OBJECTIVE: To assess the level of agreement between persons with various disabilities and their proxies in reporting community integration outcomes using the Craig Handicap Assessment and Reporting Technique (CHART). DESIGN: Reliability study. SETTING: Participants living in the community for a minimum of 6 months after onset of disability or completion of inpatient rehabilitation. PARTICIPANTS: Persons (n = 983) with disability resulting from amputation, burn, multiple sclerosis, spinal cord injury (SCI), stroke, or traumatic brain injury and their self-selected proxies. INTERVENTIONS: Telephone interview of subjects (FIM instrument, CHART); proxies (CHART). FIM instrument assesses the degree of assistance with physical and cognitive subscales; CHART measures community integration in 6 subscales: physical, cognitive, and economic independence, and mobility, social integration, and occupation. MAIN OUTCOME MEASURES: Intraclass correlation coefficients (ICCs) were used to assess participant-proxy agreement, and stepwise multiple regressions were used to identify patterns of difference in agreement based on disability type and demographic variables. RESULTS: Thirty seven of the 38 items examined for the entire sample yielded moderate to strong ICCs. Multiple regression analyses indicated that proxies overrated participants with severe functional cognitive disabilities on the mobility subscale (p < .001), overrated participants with less than a high school education on the total CHART score (p < .01), and underrated participants with SCIs on the occupation subscale (p < .01). Differences in all cases, however, were less than 6 points out of a possible score of 100 per subscale. CONCLUSIONS: Participant-proxy agreement across the 6 disability groups provided evidence in support of the inclusion of proxy data for persons with various types of disabilities in community integration research. PMID- 11494180 TI - Self-report of extent of recovery and barriers to recovery after traumatic brain injury: a longitudinal study. AB - OBJECTIVE: To examine the perspective of survivors of traumatic brain injury (TBI) regarding the extent and nature of their recovery over time. DESIGN: Inception cohort, longitudinal study. SETTING: Level I trauma center. PARTICIPANTS: One hundred fifty-seven consecutively hospitalized individuals with TBI (mean age, 36.1 yr; 80% men) with a broad range of injury severity. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Participants reported the extent of their recovery and barriers to full recovery at 1, 6, and 12 months. RESULTS: Participants reported a median return to normal at the 3 follow-up times of 65%, 80%, and 85%. After 1 month, self-reported extent of recovery correlated well with performance on the Glasgow Outcome Scale (p <.001 at 6 and 12 mo) and Wechsler Adult Intelligence Scale Performance IQ (p =.001 at 12 mo). The major reported barrier to recovery was physical difficulties, which constituted over half of the concerns at all time periods. Report of physical-related concerns decreased significantly (p =.002) over time whereas cognition-related concerns increased significantly (p =.02). CONCLUSION: Brain injury survivors consider themselves to have greater recovery than previously reported by clinicians or family members, consider physical problems a significant factor in their recovery, and appear to become more aware of cognitive impairments over time. PMID- 11494181 TI - The prevalence and risk factors of falling and fear of falling among lower extremity amputees. AB - OBJECTIVE: To estimate the falling experience and fear of falling status and to describe characteristics associated with falling and fear of falling. DESIGN: Population-based consecutive sample survey and chart review. SETTING: Two Canadian, regional, university-affiliated outpatient amputee clinics. PARTICIPANTS: The sample (n = 435; mean age, 62 +/- 15.7 yr) of community-living participants was mostly male (71%), had unilateral (below knee 73%; above knee 27%) amputations primarily for vascular (53% vs 47% nonvascular) reasons. INTERVENTION: Review patient charts and survey questionnaires to determine sociodemographic information (eg, social support), information about the amputation (eg, cause, level, problems), physical health and function (eg, pain, limitations, comorbidity), and psychologic state (depression, adaptation). MAIN OUTCOME MEASURES: Occurrence of a fall in the past 12 months and presence of a fear of falling. RESULTS: Exactly 52.4% subjects reported falling in the past year, whereas 49.2% reported a fear of falling. Logistic regression analyses revealed falling was related to having an above knee amputation (odds ratio [OR] = 2.78; 95% confidence interval [CI] = 1.71-4.51), back (OR = 1.96; 95% CI = 1.08 3.54) and joint (OR = 1.67; 95% CI = 1.01-2.74) pain, and multiple stump and prosthesis problems (OR = 3.09; 95% CI = 1.58-6.04). Having had the amputation > or = 4 years in the past was protective (OR =.53; 95% CI =.29-.89). Factors related to an increase risk of fear of falling included having to concentrate on each step while walking (OR = 4.06; 95% CI = 2.46-6.71) and having a fall in the past 12 months (OR = 1.62; 95% CI = 1.04-2.54), whereas being male (OR = 0.35; 95% CI =.21-.57) and having good to excellent perceived health (OR =.35; 95% CI =.21-.58) were protective. CONCLUSIONS: Falling and fear of falling are pervasive among amputees. Comprehensive and ongoing intervention and education should be considered. Research is required to assess the consequences of falling and fear of falling. PMID- 11494182 TI - Acupuncture and Trager psychophysical integration in the treatment of wheelchair user's shoulder pain in individuals with spinal cord injury. AB - OBJECTIVE: To determine the effectiveness of acupuncture and Trager Psychophysical Integration (a form of manual therapy) in decreasing chronic shoulder pain in wheelchair users with spinal cord injury (SCI). DESIGN: A prospective clinical trial, with subjects randomized to acupuncture or Trager treatment condition. Subjects served as their own controls by including a 5-week pretreatment baseline period and a 5-week posttreatment follow-up period. SETTING: Rehabilitation hospital research department. PARTICIPANTS: Eighteen subjects with chronic SCI and chronic shoulder pain who used manual wheelchairs as their primary means of mobility. INTERVENTION: Ten acupuncture or 10 Trager treatments over a 5-week period. MAIN OUTCOME MEASURES: Changes in performance corrected Wheelchair User's Shoulder Pain Index (PC-WUSPI) scores during baseline, treatment, and follow-up periods were assessed by using analysis of variance. RESULTS: The mean PC-WUSPI score +/- standard deviation of the 18 subjects at entry was 48.9 +/- 24.6 (range, 8.0-94). No significant change in mean PC-WUSPI scores occurred during the pretreatment baseline period. Mean PC WUSPI scores decreased significantly during the treatment period in both the acupuncture (53.4%; 23.3 points) and Trager (53.8%; 21.7 points) treatment groups. The reduced PC-WUSPI scores were maintained in both groups throughout the 5-week posttreatment follow-up period. CONCLUSION: Acupuncture and Trager are both effective treatments for reducing chronic shoulder pain associated with functional activities in persons with SCI. PMID- 11494183 TI - Physical and functional correlations of ankle-foot orthosis use in the rehabilitation of stroke patients. AB - OBJECTIVE: To examine factors associated with the use of ankle-foot orthoses (AFOs) in stroke patients undergoing rehabilitation. DESIGN: Retrospective cohort study of the frequency of AFO use. SETTING: Inpatient rehabilitation unit. PARTICIPANTS: Consecutive stroke patients (n = 423) admitted to an inpatient rehabilitation unit over a 10-year period. INTERVENTION: Discharge with AFO. MAIN OUTCOME MEASURES: Functional outcome measurement scores of patients who were and who were not prescribed an AFO were examined. The groups were compared by using admission and discharge Chedoke-McMaster Stroke Impairment Inventory (CM; each measure analyzed separately), FIMtrade mark instrument (walking, stairs, overall measures), and Berg Balance Scale scores. RESULTS: Ninety-three of the 423 patients (22%) were discharged with an AFO. Overall, they scored consistently lower than patients who were discharged without an AFO. Statistically significant differences (p <.001) were noted between AFO users and nonusers in admission and discharge scores in the arm, hand, leg, and foot components of the CM and the FIM stairs and walking component scores. Average admission and discharge Berg scores differed between the 2 groups (p =.005, p =.013, respectively). Overall FIM scores were also significantly different both at admission and discharge (p <.001, p =.025, respectively). CONCLUSION: Use of AFOs at discharge was associated with significantly lower admission and discharge CM scores of the arm, hand, leg, and foot; FIM walking and stairs scores; total FIM scores; and Berg Balance Scale scores. PMID- 11494184 TI - Gait variability and fall risk in community-living older adults: a 1-year prospective study. AB - OBJECTIVE: To test the hypothesis that increased gait variability predicts falls among community-living older adults attending an outpatient clinic. DESIGN: Prospective, cohort study. SETTING: Three outpatient geriatric clinics. PARTICIPANTS: Fifty-two community-living, ambulatory men and women aged > or = 70 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Subjects walked at a normal pace for up to 6 minutes wearing force-sensitive insoles that measured the gait rhythm on a stride-to-stride basis. Afterward, subjects reported fall status on a weekly basis for 1 year. The primary outcomes were the association between measures of the stride-to-stride fluctuations in gait rhythm and (1) subsequent falls during a 12-month follow-up period and (2) potential contributing factors. RESULTS: Almost 40% of the subjects reported falling during the 12-month follow up period. Stride time variability was 106 +/- 30 ms in subjects who subsequently fell (n = 20) and 49 +/- 4 ms in those who did not experience a fall (n = 32) during the 12-month follow-up period (p <.04). Logistic regression also showed that stride time variability predicted falls (p <.05). Stride time variability correlated significantly with multiple factors including strength, balance, gait speed, functional status, and even mental health, but these other measures did not discriminate future fallers from nonfallers. CONCLUSIONS: These findings show both the feasibility of obtaining stride-to-stride measures of gait timing in the ambulatory setting and the potential use of gait variability measures in augmenting the prospective evaluation of fall risk in community-living older adults. PMID- 11494185 TI - Gait after stroke: initial deficit and changes in temporal patterns for each gait phase. AB - OBJECTIVE: To examine which phases of the gait cycle contributed to decreased gait velocity after stroke. DESIGN: Experimental. SETTING: Inpatient rehabilitation centers. PARTICIPANTS: Forty-two patients with unilateral first stroke who were able to walk 10 meters; and 42 age- and gender-matched controls with no history of stroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Deficit and change expressed as duration (s) and proportion (%) for the 4 phases of the gait cycle at the time of admission to rehabilitation (test 1), a median of 31 days poststroke onset, and again 8 weeks later (test 2). Affected and unaffected single-limb support (SLS) and initial double-limb support (DLS) were compared. RESULTS: At tests 1 and 2, the durations of the 2 DLS and unaffected SLS phases were significantly (p <.001) longer in the stroke patients than in control subjects. No difference was found between the 2 groups for duration of affected SLS at either test time. Significant (p <.001) decreases occurred over the 8-week period in the 3 phases identified to be abnormally long at test 1. CONCLUSION: If the goal of rehabilitation is to increase gait velocity and normalize the gait pattern, treatment should focus on decreasing the DLS and unaffected SLS phases of the gait cycle. PMID- 11494186 TI - Increased H(max):M(max) ratio in community walkers poststroke without increase in ankle plantarflexion during walking. AB - OBJECTIVE: To investigate whether changes in H-reflex response at midswing and midstance are related to excessive plantarflexion during walking in community walkers poststroke compared with control subjects without stroke. DESIGN: Survey of functional walking handicap in a random sample of an annual stroke cohort followed by H-reflex and M(max) testing of a smaller sample. SETTING: Community and laboratory testing. PARTICIPANTS: Forty individuals with stroke (IWS group) completed the functional walking handicap survey, 10 of whom agreed (with 10 age matched controls) to enroll in a study of of the H(max):M(max) ratio in soleus during walking. INTERVENTION: Electromyography during treadmill walking. MAIN OUTCOME MEASURES: Functional Walking Handicap Scale, soleus H(max):M(max) ratio, and the ankle joint's angle of displacement. RESULTS: Nine of the 10 stroke patients were community walkers. All had significantly (p <.05) more variable ankle movement during walking than the controls. The H(max):M(max) ratio was significantly (p <.01) increased in the IWS group because of a decrease in M(max) response without significant (p >.05) increase in H(max) response. CONCLUSIONS: Individuals with community-level walking ability after stroke have significantly (p <.05) less repeatability of ankle joint movement than controls at both midswing and midstance. Simultaneous soleus H(max) and M(max) testing showed a significant (p <.01) reduction in the H(max) and H(max):M(max) ratio at midswing in controls only. This inhibition at midswing was lost by the IWS group without significant increase in H(max), suggesting that central synaptic excitability was within the normal range, and possibly accounting for the absence of excessive ankle plantarflexion during walking in the IWS group with community level walking ability. PMID- 11494187 TI - Recovery from moderate aphasia in the first year poststroke: effect of type of therapy. AB - OBJECTIVES: (1) To determine whether 2 model-based remediation programs affect writing performance in unselected subjects with moderate aphasia and whether there is consequent improvement in everyday life, and (2) to interpret the potential changes observed by recourse to a theoretical model. DESIGN: Consecutive sample, multiple baseline, within subject crossover study. SETTING: Ambulatory care units. PARTICIPANTS: Eight subjects with moderate aphasia from 6 to 12 months postonset. INTERVENTION: A standardized test for reading and writing skills was given at the beginning and the end of each therapy program and 1 month after therapy stopped. MAIN OUTCOME MEASURES: Functional outcome measures were the Communicative Abilities in Daily Living (CADL) test and subtests from standardized aphasia assessment. RESULTS: After the 2 programs, there was improved writing performance, which was maintained after therapy stopped. Patterns of improvement corresponded to each of the 2 programs. Learning transfer was observed on the CADL test and functional writing, but gains on oral language were limited. Only 1 program was effective for 6 of the 8 patients. CONCLUSION: Specific rehabilitation programs aid recovery from aphasic symptoms from 6 to 12 months postonset. Individual response is linked to type of treatment. The interpretation is linked to a model-based description of aphasic symptoms and mechanisms of functional recovery. PMID- 11494188 TI - Contraction of the pelvic floor muscles during abdominal maneuvers. AB - OBJECTIVE: To determine whether voluntary abdominal muscle contraction is associated with pelvic floor muscle activity. DESIGN: Pelvic floor muscle activity was recorded during contractions of the abdominal muscles at 3 different intensities in supine and standing positions. SETTING: Research laboratory. PARTICIPANTS: Six women and 1 man with no histories of lower back pain. INTERVENTION: Not applicable. MAIN OUTCOME MEASURES: Electromyographic activity of the pelvic floor muscles was recorded with surface electrodes inserted into the anus and vagina. These recordings were corroborated by measurements of anal and vaginal pressures. Gastric pressure was recorded in 2 subjects. RESULTS: Pelvic floor muscle electromyography increased with contraction of the abdominal muscles. With strong abdominal contraction, pelvic floor muscle activity did not differ from that recorded during a maximal pelvic floor muscle effort. The pressure recordings confirmed these data. The increase in pressure recorded in the anus and vagina preceded the pressure in the abdomen. CONCLUSIONS: In healthy subjects, voluntary activity in the abdominal muscles results in increased pelvic floor muscle activity. The increase in pelvic floor pressure before the increase in the abdomen pressure indicates that this response is preprogrammed. Dysfunction of the pelvic floor muscles can result in urinary and fecal incontinence. Abdominal muscle training to rehabilitate those muscles may be useful in treating these conditions. PMID- 11494189 TI - Back and abdominal muscle function during stabilization exercises. AB - OBJECTIVES: To assess the paraspinal and abdominal muscle activities during different therapeutic exercises and to study how load increment produced by varying limb movements and trunk positions could affect these muscle activities. DESIGN: A cross-sectional study comparing muscle activities between men and women. SETTING: Rehabilitation clinic in university hospital. PARTICIPANTS: Twenty-four healthy volunteers (14 women, 10 men) aged 21 to 39 years. INTERVENTIONS: Subjects performed 16 different therapeutic exercises commonly used to treat low back pain. MAIN OUTCOME MEASURES: Surface electromyography was recorded from the paraspinal (T9, L5) and abdominal (rectus abdominis, obliquus externus) muscles during these exercises. Average electromyographic amplitudes obtained during the exercises were normalized to the amplitude in maximal voluntary contraction (% MVC) to produce interindividually comparable muscle activity assessments. RESULTS: Mean average normalized electromyographic amplitudes (% MVC) of the exercises were below 50% MVC. At L5 level, the multifidus muscle activities were significantly higher (p <.05) in women than in men, whereas no significant difference was found at T9 level. Similarly, rectus abdominis and obliquus externus activities were significantly higher (p <.001, p <.05) in women than in men. Load increment in hands or unbalanced trunk and limb movements produced higher paraspinal and abdominal muscle activities (p <.05). CONCLUSIONS: Simple therapeutic exercises are effective in activating both abdominal and paraspinal muscles. By changing limb and trunk positions or unbalancing trunk movements, it is possible to increase trunk muscle activities. Women were better able to activate their stabilizing trunk muscles than men; but it is also possible that men, having a much higher degree of strength on maximal contraction, only need to activate a smaller amount of that maximum to perform a similar activity. PMID- 11494190 TI - Short-term outcomes of a back school program for chronic low back pain. AB - OBJECTIVE: To assess the short-term outcome of a back school program for patients suffering from chronic, nonspecific low back pain (LBP). DESIGN: Quasi experimental cohort study with a waiting list control group. SETTING: Dutch rehabilitation department. PARTICIPANTS: Experimental group (n = 14) participating in the back school program and a waiting list control group (n = 10). INTERVENTION: A back school program aimed to achieve optimal functional capacity and functional health status by teaching participants to react appropriately to overload signals. MAIN OUTCOME MEASURES: Functional capacity assessed by the RAND-36 instrument; functional health status assessed by the Roland-Morris Disability Questionnaire; and static and dynamic lifting capacity, endurance, and range of motion assessed by objective measures. RESULTS: The experimental group significantly improved in functional capacity and functional health status, with a large power, likely attributable to adequate reactions to signals of overload. Significant differences existed between the 2 groups, with large powers for the main outcomes. CONCLUSION: The back school program improved the functional capacity and functional health status of patients with chronic, nonspecific LBP. PMID- 11494191 TI - The Montreal rehabilitation performance profile: a task-analysis approach to quantify stair descent performance in children with intellectual disability. AB - OBJECTIVE: To develop a clinical tool to quantify stair descent performance in children with moderate to severe intellectual disability. DESIGN: Item identification, measurement construction, and basic testing of reliability. SETTING: School for children with intellectual disability. PARTICIPANTS: A sample of convenience; 18 children (age range, 5-9 yr) with moderate to severe intellectual disability, but who were ambulatory. INTERVENTION: The Montreal Rehabilitation Performance Profile (MRPP) measurement tool, which has 4 perceptual-motor elements: movement form, time requirement, proprioceptive cues, and external cues. MAIN OUTCOME MEASURE: Identification of perceptual-motor elements that underlie skilled stair descent and that are directly quantified from standard video recording. RESULTS: Interrater reliability measured by percentage agreement (kappa statistic) was high for 3 elements (80%-100%; kappa =.63) and good for the fourth (67%; kappa =.52). The 4 MRPP elements are plotted on cartesian axes to yield (1) the magnitude of the contribution of each independent element to total performance outcome, and (2) a global measurement of level of functional skill, the Performance Composite Score (PCS). CONCLUSIONS: The MRPP, and associated PCS, provide a valid measure of functional stair descent skill that does not rely on cognitive understanding of the process. This tool could be adapted to measure functional capacity in other clinical populations, including geriatric clients. PMID- 11494192 TI - Association among neuromuscular and anatomic measures for patients with knee osteoarthritis. AB - OBJECTIVE: To investigate neuromuscular and anatomic factors involved in varus gonarthrosis by identifying measures associated with degenerative changes. DESIGN: Descriptive study. Individual measures that explained substantial portions of the variability in ratings of knee joint-degenerative changes in patients with knee medial compartment osteoarthritis. SETTING: Outpatient orthopedic clinic and biomechanics and muscular assessment laboratory. PATIENTS: Volunteer sample of 20 subjects (age range, 59 +/- 9 yr) with no history of neurologic disease. INTERVENTION: Not applicable. MAIN OUTCOME MEASURES: We assessed degenerative changes, varus alignment, standing balance, and knee proprioception. Weight-bearing radiographs were used to assess the extent of degenerative changes and the degree of varus alignment. Single-limb standing balance control was assessed through tests performed on a force platform. Knee proprioception was assessed with an isokinetic dynamometer, using a joint angle replication test. RESULTS: Forward-stepwise multiple linear regression indicated that the extent of degenerative changes could be best predicted from a linear combination of the independent variables, varus alignment, and standing balance (R =.80, F(2,17) = 14.81, p =.0002). Sixty-four percent of the variability in ratings of degenerative changes was explained by alignment and standing balance measures (37% by varus alignment, 27% by standing balance). Alignment and balance measures were poorly correlated (r =.12, p =.63), further suggesting that they provided different information about gonarthrosis. CONCLUSIONS: Although varus alignment is widely accepted as a clinically important factor in gonarthrosis, and is the focus of many treatment efforts, our results suggest that objective measures of standing balance are also important. As a result, the potential impact of rehabilitation to improve the control of standing balance should be further evaluated in this patient population. PMID- 11494193 TI - Monitoring in-shoe plantar pressures, temperature, and humidity: reliability and validity of measures from a portable device. AB - OBJECTIVE: To investigate the reliability and validity of measures obtained from a portable electronic device used to monitor changes in plantar pressure, temperature, and humidity that occur within the shoe during prolonged activity. DESIGN: Descriptive study comparing electronic sensor output with criterion values. SETTINGS: Indoor level walkway for pressure data; uncontrolled, outdoor environment for step count data; enclosed environmental control chamber for humidity and temperature data. PARTICIPANTS: Convenience sample of 4 healthy, sensate subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Correlations between sensor output and criterion measures were determined for pressure and temperature data. The absolute differences between sensor output and criterion values of temperature, humidity, and step count were also determined. RESULTS: Pressure measurements from electronic sensors correlated highly with criterion values (r > or =.82), both before and after prolonged use. Relative humidity sensor output were within 5% of hygrometer values. In-shoe temperature data correlated highly with criterion values (r > or =.99), and differed from known temperatures by.50 degrees +/-.84 degrees C and.96 degrees +/- 1.56 degrees C at the forefoot and heel, respectively. Electronic step counts recorded at the central forefoot were within 1 step of visual step counts. Pressure tracings obtained from the device during different weight-bearing activities revealed qualitatively distinct pressure patterns. CONCLUSION: The device provides valid and reliable measures of in-shoe plantar pressures, temperature, and humidity during prolonged activity. PMID- 11494194 TI - Reliability of stationary dynamometer muscle strength testing in community dwelling older adults. AB - OBJECTIVES: To determine the 1-week test-retest reliability of stationary dynamometer scores in the measurement of muscle strength in older adults and to determine the reliability of composite scores obtained by combining right and left lower limb strength scores for each muscle group. DESIGN: In separate sessions, 1 therapist performed repeated measurements of muscle force production. SETTING: Outpatient physical therapy clinic of a large teaching hospital. PARTICIPANTS: A convenience sample of 25 volunteers aged 70 to 87 years residing independently in the community and who did not have significant health problems. INTERVENTION: On 2 separate occasions, 1 week apart, bilateral isometric force measurements were obtained for the flexor and extensor muscle groups of the ankle, knee, and hip joints. MAIN OUTCOME MEASURES: For test-retest reliability of individual and composite scores, the intraclass correlation coefficients (ICCs) and 90% confidence intervals were determined. RESULTS: The mean scores for ankle dorsiflexion, knee flexion and extension, and hip flexion exhibited excellent reliability with ICCs ranging from.90 to.76 for the individual lower limb scores and.91 to.84 for the composite scores. Scores for the remaining muscle groups exhibited good reliability with ICCs ranging from.74 to.71 for the composite scores. CONCLUSION: The stationary dynamometer is a reliable tool to use in determining lower limb muscle force production in elderly adults. PMID- 11494195 TI - Potential role of mental practice using motor imagery in neurologic rehabilitation. AB - For many patients with damage to the central nervous system (CNS), execution of motor tasks is very difficult, sometimes impossible, even after early participation in an active rehabilitation program. Several investigators have recently proposed that mental practice could be used by these patients as a therapeutic tool to improve their performance of motor functions, yet very little empirical work addresses this issue directly. This article discusses the rationale for investigating mental practice as a means of promoting motor recovery in patients with a neurologic disorder. We first present evidence supporting the existence of a similarity between executed and imagined actions using data from psychophysical, neurophysiologic, and brain imaging studies. This parallel is then extended to the repetition of movements during physical and mental practice of a motor skill. Finally, a new model is proposed to emphasize the key role of motor imagery as an essential process of mental practice, and also to stimulate additional research on this type of training in the rehabilitation of patients with motor impairments of cerebral origin. PMID- 11494196 TI - Thoracic disk herniation presenting as abdominal and pelvic pain: a case report. AB - We report the case of a young woman who presented with a 2-month history of severe abdominal and pelvic pain. The past history was significant for a fall from a bicycle 1 week before the onset of her pain. Physical examination was remarkable for periumbilical tenderness. Work-up including pelvic sonogram and diagnostic laparoscopy suggested endomyometritis. The pain was minimally relieved by nonsteroidal anti-inflammatory drugs and narcotic analgesics. Thoracic spine magnetic resonance imaging (MRI) revealed a large disk herniation at the T9-10 level compressing the spinal cord. The patient subsequently underwent T9-10 diskectomy and laminectomy with dramatic relief of her symptoms. Postoperative rehabilitation hastened her functional improvement. This is a rare case of symptomatic thoracic disk herniation after trauma presenting as abdominal and pelvic pain. Physicians should be aware of this unusual presentation of thoracic disk herniation to avoid invasive diagnostic procedures. PMID- 11494197 TI - Ataxia from lithium toxicity successfully treated with high-dose buspirone: a single-case experimental design. AB - Injury to the cerebellum commonly results in clumsiness or uncoordinated movement, which is referred to as ataxia. The severity of ataxia varies according to the extent of the lesion. Severe ataxia usually restricts activities of daily living, impairs mobility, and increases level of disability. Recent studies investigating use of serotonin agonists in the treatment of ataxia have produced mixed results; however, buspirone with an affinity specific to the 5 hydroxytryptamine(1A) subreceptors has shown promise. In this brief report, we use a prospective, open, single-case experimental design to describe substantial subjective and objective dose-dependent improvement of ataxia after unusually high doses of buspirone taken by a patient whose severe ataxia was due to lithium toxicity. PMID- 11494199 TI - Preoperative assessment of the thoracic surgery patient: introduction. PMID- 11494200 TI - Preoperative assessment of the thoracic surgery patient: pulmonary function testing. AB - Tests of pulmonary function before thoracic surgery can help to assess the risk of perioperative morbidity and mortality, and are the basis for estimating remaining lung function after resection of lung tissue. Testing has evolved over the past 50 years from reliance on the maximum breathing capacity to a range of studies including spirometry, and measurement of lung volume, diffusing capacity, and arterial blood gases, and the interpretation of these in conjunction with radionuclide scanning. The surgeon must consider both the early postoperative decrement in function and the level to which the patient is expected to recover. Although it is difficult to establish absolute limiting values, and current surgical techniques are blurring the boundary further, the reported experience underlying current guidelines is reviewed. PMID- 11494201 TI - Evaluation and management of patients with pulmonary disease before thoracic and cardiovascular surgery. AB - The risks of respiratory complications after thoracic and cardiovascular surgeries are particularly high for patients with chronic pulmonary disease and are associated with prolonged hospital stays and increased mortality. The primary goals of preoperative management are to identify risk factors and institute interventions likely to reduce subsequent postoperative pulmonary complications. Smoking, symptomatic obstructive lung disease, respiratory infection, obesity, and malnutrition are all potentially modifiable risk factors. Chest physiotherapy is indicated in all patients regardless of risk factor profile. Providing a thoughtfully designed, multifaceted course of preoperative care can result in a clinically significant reduction in postoperative morbidity and mortality, particularly if instituted well in advance of surgery. PMID- 11494202 TI - Cardiopulmonary exercise testing in the preoperative assessment for lung resection surgery. AB - Whereas pulmonary function tests (PFTs) initially identify high-risk pulmonary patients being evaluated for lung resection surgery, other diagnostic modalities, including cardiopulmonary exercise testing (CPET) and/or split function studies, are then necessary for a more accurate assessment. CPET including VO2max have emerged as integral components of a step approach for the physiologic assessment for lung resection surgery. Increasingly, CPET is being used because it provides the best index of functional capacity and global O2 transport (VO2max) as well as estimating both cardiac and pulmonary reserves not available from other modalities. CPET permits the detection of clinically occult heart disease and provides a more reliable estimate of functional capacity postoperatively compared with PFTs, which routinely overestimate functional loss after lung resection. Currently, though split function studies are clearly established and have traditionally been used before CPET in preoperative decision analysis, recent work favors using CPET including VO2max before split function studies because VO2max % predicted is a good independent predictor of risk. Importantly, both studies are complementary and optimize assessment of surgical risk; this is particularly valuable for borderline patients, so that opportunity for curative resection is not denied. PMID- 11494203 TI - Functional imaging before pulmonary resection. AB - The evaluation of patients before lung resections requires functional imaging. We review the current role of functional imaging in the preoperative evaluation of candidates for lung volume reduction surgery (LVRS) and lung cancer resection. Perfusion and ventilation lung scintigraphy methods as well as computed tomography techniques used to predict postoperative lung function are presented. The areas of current investigation are also described. PMID- 11494204 TI - Cardiac risk assessment in noncardiac thoracic surgery. AB - Preoperative cardiac risk assessment for noncardiac thoracic surgery is limited by the lack of data specific to this type of surgery, especially prospective, controlled data. However, the value of clinical predictors in determining accurate postoperative cardiac outcomes is a reliable tool. Thus, the approach is similar to traditional cardiac risk assessment for noncardiac surgery. The essential elements of cardiovascular evaluation as it pertains to noncardiac thoracic surgery are reviewed with a specific focus on coronary artery disease, perioperative arrhythmias, and selected topics relevant to noncardiac thoracic surgery. The core recommendations of the clinical guidelines by the American College of Cardiology and American Heart Association are discussed in the context of noncardiac thoracic surgery. PMID- 11494205 TI - The neurologic complications of cardiac surgery: introduction. PMID- 11494206 TI - Evaluation of neurologic assessment and outcomes in cardiac surgical patients. AB - It is well recognized that cardiac surgery with cardiopulmonary bypass can potentially induce a wide spectrum of central nervous system (CNS) sequelae. Our awareness of the prevalence of these CNS complications is dependent on the validity of the available diagnostic methods. Current assessment methods designed to detect both focal and diffuse cerebral ischemia include neurologic examination, imaging techniques, biochemical markers, neuropsychologic assessment, and patient perceived outcomes. These techniques vary in their sensitivity and specificity, as well as feasibility for use in everyday clinical practice. There are currently only limited standardized methodologic guidelines for the assessment of CNS complications after cardiac surgery, which has resulted in considerable interstudy variability in the identification and reporting of outcomes. The application of clearly definable endpoints for reporting of CNS outcomes would be beneficial. The wealth of available data suggests that the incidence of overt CNS injury such as stroke has declined since the 1980s and is now approximately 2%, whereas evidence suggests that up to one third of cardiac surgery patients experience postoperative cognitive deficits. One of the clear strengths of the current era is the recognition of CNS injury associated with cardiac surgery, and the quest to improve our understanding of these outcomes. The application of more uniform assessment and reporting practices is surely vital to the continued advancement of cardiac surgery. PMID- 11494207 TI - Beating heart surgery or conventional CABG: are neurologic outcomes different? AB - Although there has been much debate about the causes of neurologic complications associated with coronary artery bypass grafting (CABG), there is good evidence linking such complications with some of the pathophysiologic changes associated with use of conventional cardiopulmonary bypass (CPB). Several studies indicate that it is possible to significantly lower risk of stroke and other central nervous system (CNS) morbidity in patients undergoing CPB for CABG by application of selected techniques and equipment modifications. The resurgence of interest in coronary revascularization by using beating heart surgery (BHS) offers a unique opportunity to evaluate neurologic outcome independent of CPB. Currently, BHS would appear to significantly reduce morbidity in the elderly and to decrease the costs and resource use in coronary revascularization patients. It is hoped that by understanding the mechanisms of CNS injury associated with CABG, techniques can be developed to decrease the risk of neurologic injury associated with coronary revascularization, whether or not CPB is used. Definitive conclusions regarding outcomes after best practice CPB or BHS await large-scale, risk stratisfied multicenter trials. PMID- 11494208 TI - Importance of blood pressure regulation in maintaining adequate tissue perfusion during cardiopulmonary bypass. AB - Patients undergoing surgery with the aid of cardiopulmonary bypass (CPB) have an incidence of end-organ dysfunction, caused by embolization, regional hypoperfusion, or some combination of the two. In this article, we attempt to define the effect of mean arterial pressure (MAP) during CPB on postoperative end organ function. Although early studies reported that cerebral perfusion during hypothermic CPB is independent of MAP, recent laboratory and clinical reports have shown a positive slope in the MAP versus cerebral blood flow relationship. In clinical studies, patients who had higher MAPs during CPB had a lower incidence of cardiac and neurologic complications, as well as late neurocognitive abnormalities compared with patients with lower MAPs. Improving collateral flow in the setting of cerebral embolization has been postulated as the main mechanism for the improved neurologic outcomes in the high MAP groups. Higher perfusion pressure during CPB affects regional blood flow to the kidneys and visceral organs. However, the lower autoregulatory limits of perfusion to abdominal organs differ from the limits to the brain. Enhanced visceral perfusion during CPB is best achieved by increasing perfusion pressure via increases in perfusion flow rates rather than by using peripheral vasoconstriction alone. In conclusion, it is clear that maintenance of a high MAP during CPB may have a significant impact in protecting the brain and abdominal organs, particularly in the subset of patients at high risk for embolization and end-organ dysfunction. PMID- 11494209 TI - Cerebral hyperthermia and cardiac surgery: consequences and prevention. AB - Temperature management during cardiac surgery deserves considerable attention because it has broad effects, altering virtually every physiologic process, including oxygen demand, blood flow, cardiac output, and coagulation. Temperature is also important in cardiac surgery because virtually all patients undergo significant temperature change. These changes can be unique in mammalian physiology both with regard to their magnitude and rate of change. Furthermore, cardiac surgical patients may be uniquely vulnerable to the effects of temperature. Because of vascular disease and embolization, many patients are at risk for cerebral ischemia. Additionally, their cardiac, pulmonary, and renal reserve is typically limited; and there is risk for perioperative bleeding. Patient temperature can affect all of these processes and has its greatest effect on those who are physiologically most fragile. An appreciation for temperature management is also compelling because, unlike new technologies, procedures, or drugs; temperature management is simple, practical, applicable to every patient, and can be performed with very little cost. This article will show why cerebral hyperthermia should be avoided in cardiac surgery. Second, it will discuss why it occurs and the management steps that may prevent it. Finally, we will highlight recent discussion of postoperative hyperthermia and speculate as to its origin and relevance. PMID- 11494210 TI - Approaches to reduce neurologic complications during cardiac surgery. AB - As outcome measures of cardiac surgery are carefully analyzed, neurologic outcomes are a prominent determinant of overall outcome. Sensitive measures of pre- and postoperative neuropsychologic performance and intraoperative emboli reveal a risk group of patients at an advanced age, with severe generalized atherosclerosis who require cardiac surgery. By using the results of observational studies, we have developed a protocol that uses innovative intraoperative techniques to minimize injury and thus improve outcome. PMID- 11494211 TI - Management of patients with concomitant coronary and carotid vascular disease. AB - Stroke is a major complication of coronary artery bypass graft (CABG) surgery. Carotid stenosis is an important cause of stroke in certain CABG patients. Randomized trials have revealed that carotid endarterectomy (CEA) is clearly indicated in non-CABG patients with symptomatic severe carotid stenosis. CEA is also indicated in patients with symptomatic moderate stenosis and asymptomatic severe stenosis if the predicted incidence of perioperative morbidity and mortality is low. Therapeutic options for patients with concomitant coronary and carotid disease include CABG alone, CABG plus CEA, and CABG plus carotid stenting. In this article we discuss each of these management techniques in detail, and make recommendations regarding the preferred approach in specific patient populations. PMID- 11494212 TI - Process performance models in the optimization of multiproduct protein production plants. AB - In this work we propose a model that simultaneously optimizes the process variables and the structure of a multiproduct batch plant for the production of recombinant proteins. The complete model includes process performance models for the unit stages and a posynomial representation for the multiproduct batch plant. Although the constant time and size factor models are the most commonly used to model multiproduct batch processes, process performance models describe these time and size factors as functions of the process variables selected for optimization. These process performance models are expressed as algebraic equations obtained from the analytical integration of simplified mass balances and kinetic expressions that describe each unit operation. They are kept as simple as possible while retaining the influence of the process variables selected to optimize the plant. The resulting mixed-integer nonlinear program simultaneously calculates the plant structure (parallel units in or out of phase, and allocation of intermediate storage tanks), the batch plant decision variables (equipment sizes, batch sizes, and operating times of semicontinuous items), and the process decision variables (e.g., final concentration at selected stages, volumetric ratio of phases in the liquid-liquid extraction). A noteworthy feature of the proposed approach is that the mathematical model for the plant is the same as that used in the constant factor model. The process performance models are handled as extra constraints. A plant consisting of eight stages operating in the single product campaign mode (one fermentation, two microfiltrations, two ultrafiltrations, one homogenization, one liquid-liquid extraction, and one chromatography) for producing four different recombinant proteins by the genetically engineered yeast Saccharomyces cerevisiae was modeled and optimized. Using this example, it is shown that the presence of additional degrees of freedom introduced by the process performance models, with respect to a fixed size and time factor model, represents an important development in improving plant design. PMID- 11494213 TI - Evaluation of photobioreactor heat balance for predicting changes in culture medium temperature due to light irradiation. AB - Microalgal photosynthesis requires appropriate culture medium temperatures to achieve high photosynthetic performance and to maintain production of a high quality biomass product. Enclosed systems, such as our conical, helical tubular photobioreactor (HTP), can accomplish high photosynthetic efficiency and the small amount of culture medium used by these systems means that the culture medium temperature may be effectively controlled. On the other hand, because a high ratio of surface area to culture medium volume leads to rapid heating under the illumination condition and substantial heat loss at night, maintaining a suitable culture medium temperature is necessary to achieve efficient, commercially practical biomass production. In order to predict changes in the culture medium temperature caused by changes in solar irradiance and ambient temperature, it is necessary to understand the heat balance within the photobioreactor. We therefore investigated the heat balance in three major parts (photostage, degasser, and helical heat exchanger) of our conical HTP, analyzed the time-dependent changes in medium temperature at various room temperatures and radiant energy inputs, and predicted changes in the culture medium temperature based on the characteristics of heat transfer among the three parts. Using this model, the predicted changes in culture medium temperature were very similar to the changes observed experimentally in the laboratory and under field conditions. This means that by calculating the time-dependent changes in the culture medium temperature, based on measurements of solar energy input and ambient temperature, we should be able to estimate the energy required to maintain the culture medium temperature within a range where photosynthetic performance of microalgae is high. PMID- 11494214 TI - Effects of nonionic surfactants on the UV/visible absorption of bacterial cells. AB - Nonionic surfactants are used in a number of different microbiological applications, including solubilization of cell membranes, washing bacterial cultures prior to experimentation, and enhancing biodegradation of low-solubility compounds. An important consideration in these applications is the potential for the surfactant to alter the cell membrane. One potential means to monitor the impact of surfactants on the bacterial cell membrane is through monitoring the absorbance spectrum of the bacterial suspension. This is due to the colloidal nature of bacteria, where the absorbance of a bacterial suspension is related to the size and refractive index of the bacterial cells. Through a systematic study it was shown that there can be a significant change in the bacterial absorbance spectrum due to the presence of nonionic surfactants, with the effect a function of surfactant structure and concentration, solution ionic strength and cation valence. The effects were most pronounced with Na(+) as the cation, with surfactants having mid-range hydrophile-lipophile balance (HLB) values, and with surfactant concentrations above the CMC. The results indicate that measurement of the absorbance spectrum of bacterial cultures can provide a means to monitor the effects of nonionic surfactants on the bacterial cell membrane. In addition, depending on the specific application, appropriate selection of surfactant structure and media composition can be made to enhance or minimize the effects. PMID- 11494215 TI - Optimization of carbohydrate fatty acid ester synthesis in organic media by a lipase from Candida antarctica. AB - The effect of solvents and solvent mixtures on the synthesis of myristic acid esters of different carbohydrates with an immobilized lipase from C. antarctica was investigated. The rate of myristyl glucose synthesized by the enzyme was increased from 3.7 to 20.2 micromol min(-1) g(-1) by changing the solvent from pure tert-butanol to a mixture of tert-butanol:pyridine (55:45 v/v), by increasing the temperature from 45 degrees C to 60 degrees C, and by optimizing the relative amounts of glucose, myristic acid, and the enzyme preparation. Addition of more than 2% DMSO to the tert-butanol:pyridine system resulted in a reduction of enzyme activity. Lowering the water content of the enzyme preparation below 0.85% (w/w) resulted in significant decreases in enzyme activity, while increasing the water content up to 2.17% (w/w) did not significantly affect the enzyme activity. The highest yields of myristyl glucose were obtained when an excess of unsolubilized glucose was present in the reaction system. In this case, all of the initially solubilized and a significant amount of the initially unsolubilized glucose was converted to the ester within 24 h of incubation, resulting in a myristyl glucose concentration of 34 mg/mL(-1). Myristic acid esters of fructose (22.3 micromol min(-1) g(-1)), alpha-D-methyl glucopyranoside (26.9 micromol min(-1) g(-1)) and maltose (1.9 micromol min(-1) g(-1)) could also be prepared using the tert-butanol:pyridine solvent system. No synthesis activity was observed with maltotriose, cellobiose, sucrose, and lactose as substrate. PMID- 11494216 TI - High-level expression and stabilization of recombinant human chitinase produced in a continuous constitutive Pichia pastoris expression system. AB - A continuous fermentation process has been developed in Pichia pastoris (P. pastoris) with the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter in order to produce large quantities of recombinant human chitinase (rh-chitinase) for preclinical studies as a potential high-dose antifungal drug. Expression levels of about 200 to 400 mg/L have been demonstrated in fed-batch fermentations using strains with either the traditional methanol-inducible or the constitutive GAP promoter. Proteolytic degradation of the enzyme was typically seen in fed batch fermentations. Continuous production of the enzyme by P. pastoris with the GAP promoter was demonstrated in a 1.5-L working volume fermentor using either glucose or glycerol as the carbon source. The fermentation could be extended for >1 month with a steady-state protein concentration of approximately 300 mg/L. Cell densities were >400 g/L wet cell weight (WCW) (approximately 100 g/L dry cell weight [DCW]) at a dilution rate (D) of 0.83 day(-1) or 1.2 volume exchanges per day (VVD). No proteolytic degradation of the enzyme was seen in the continuous fermentation mode. PMID- 11494217 TI - Factors affecting alpha,-1,2 glucooligosaccharide synthesis by Leuconostoc mesenteroides NRRL B-1299 dextransucrase. AB - The optimization of alpha-1,2 glucooligosaccharide (GOS) synthesis from maltose and sucrose by Leuconostoc mesenteroides NRRL B-1299 dextransucrase was achieved using experimental design and consecutive analysis of the key parameters. An increase of the pH of the reaction from 5.4 to 6.7 and of the temperature from 25 to 40 degrees C significantly favored alpha-1,2 GOS synthesis, thanks to a significant decrease of the side reactions, i.e., dextran and leucrose synthesis. These positive effects were not sufficient to compensate for the decrease of enzyme stability caused by the use of high pH and temperature. However, the critical parameters were the sucrose to maltose concentration ratio (S/M) and the total sugar concentration (TSC). Alpha1,2 GOS synthesis was favored at high S/M ratios. But using these conditions also led to an increase of side reactions which could be modulated by choosing the appropriate TSC. Finally, with S/M = 4 and TSC = 45% w/v, dextran and leucrose productions were limited and the final alpha-1,2 GOS yield reached 56.7%, the total GOS yield being 88%. PMID- 11494218 TI - A priori analysis of metabolic flux identifiability from (13)C-labeling data. AB - The (13)C-labeling technique was introduced in the field of metabolic engineering as a tool for determining fluxes that could not be found using the 'classical' method of flux balancing. An a priori flux identifiability analysis is required in order to determine whether a (13)C-labeling experiment allows the identification of all the fluxes. In this article, we propose a method for identifiability analysis that is based on the recently introduced 'cumomer' concept. The method improves upon previous identifiability methods in that it provides a way of systematically reducing the metabolic network on the basis of structural elements that constitute a network and to use the implicit function theorem to analytically determine whether the fluxes in the reduced network are theoretically identifiable for various types of real measurement data. Application of the method to a realistic flux identification problem shows both the potential of the method in yielding new, interesting conclusions regarding the identifiability and its practical limitations that are caused by the fact that symbolic calculations grow fast with the dimension of the studied system. PMID- 11494219 TI - Simple and robust method for estimation of the split between the oxidative pentose phosphate pathway and the Embden-Meyerhof-Parnas pathway in microorganisms. AB - The flux through the oxidative pentose phosphate (PP) pathway was estimated in Bacillus clausii, Saccharomyces cerevisiae, and Penicillium chrysogenum growing in chemostats with [1-(13)C]glucose as the limiting substrate. The flux calculations were based on a simple algebraic expression that is valid irrespective of isotope rearrangements arising from reversibilities of the reactions in the PP pathway and the upper part of the Embden-Meyerhof-Parnas pathway. The algebraically calculated fluxes were validated by comparing the results with estimates obtained using a numerical method that includes the entire central carbon metabolism. Setting the glucose uptake rate to 100, the algebraic expression yielded estimates of the PP pathway flux in B. clausii, S. cerevisiae, and P. chrysogenum of 20, 42, and 75, respectively. These results are in accordance with the results from the numerical method. The information on the labeling patterns of glucose and the proteinogenic amino acids were obtained using gas chromatography / mass spectrometry, which is a very sensitive technique, and therefore only a small amount of biomass is needed for the analysis. Furthermore, the method developed in this study is fast and readily accessible, as the calculations are based on a simple algebraic expression. PMID- 11494220 TI - Biocatalysis of isomerization of 1,2-benzisoxazole by bovine serum albumin in an organic solvent. AB - The catalytic properties of bovine serum albumin (BSA) have been studied rather extensively. The protein has shown catalytic activities toward a variety of reactions in aqueous media. However, there appears to be no reports of reactions catalyzed exclusively in organic media. In this study, we report the catalytic property of BSA on the isomerization of 1,2-benzisoxazole in an organic solvent. Kinetics were performed using a continuous flow system in a spectrophotometer at 330 nm. The catalysis showed a Michaelis-Menten behavior with an estimated catalytic efficiency (k(cat)/K(M)) of 8.77 M(-1)S(-1). The influence of water content and inhibitors was also studied. The activity of BSA in acetonitrile is approximately 20% of that in water under similar conditions. Inhibition studies show that the active site may be the pyridoxal binding site involving Lys-220. PMID- 11494221 TI - Metabolic-flux analysis of continuously cultured hybridoma cells using (13)CO(2) mass spectrometry in combination with (13)C-lactate nuclear magnetic resonance spectroscopy and metabolite balancing. AB - Protein production of mammalian-cell culture is limited due to accumulation of waste products such as lactate, CO(2), and ammonia. In this study, the intracellular fluxes of hybridoma cells are measured to determine the amount by which various metabolic pathways contribute to the secretion of waste products derived from glucose. Continuously cultured hybridoma cells are grown in medium containing either 1-(13)C-, 2-(13)C-, or 6-(13)C-glucose. The uptake and production rates of amino acids, glucose, ammonia, O(2), and CO(2) as well as the cellular composition are measured. In addition, the (13)C distribution of the lactate produced and alanine produced by the hybridomas is determined by (1)H-NMR spectroscopy, and the (13)CO(2)/(12)CO(2) ratio is measured by on-line mass spectrometry. These data are used to calculate the intracellular fluxes of the glycolysis, the pentose phosphate pathway, the TCA cycle, and fluxes involved in amino acid metabolism. It is shown that: (i) approximately 20% of the glucose consumed is channeled through the pentose shunt; (ii) the glycolysis pathway contributes the most to lactate production, and most of the CO(2) is produced by the TCA cycle; (iii) the pyruvate-carboxylase flux is negligibly small; and (iv) the malic-enzyme flux is estimated to be 10% of the glucose uptake rate. Based on these flux data suggestions are made to engineer a more efficient glucose metabolism in mammalian cells. PMID- 11494222 TI - Solar energy conversion by green microalgae: a photosystem for hydrogen peroxide production. AB - A photosystem for solar energy conversion, comprised of a culture of green microalgae supplemented with methyl viologen, is proposed. The capture of solar energy is based on the Mehler reaction. The reduction of methyl viologen by the photosynthetic apparatus and its subsequent reoxidation by oxygen produces hydrogen peroxide. This is a rich-energy compound that can be used as a nonpollutant and efficient fuel. Four different species of green microalgae, Chlamydomonas reinhardtii (21gr) C. reinhardtii (CW15), Chlorella fusca, and Monoraphidium braunii, were tested as a possible biocatalyst. Each species presented a different efficiency level in the transformation of energy. Azide was an efficient inhibitor of the hydrogen peroxide scavenging system while maintaining photosynthetic activity of the microalgae, and thus significantly increasing the production of the photosystem. The strain C. reinhardtii (21gr), among the species studied, was the most efficient with an initial production rate of 185 micromol H(2)O(2)/h x mg Chl and reaching a maximum of 42.5 micromol H(2)O(2)/mg Chl when assayed in the presence of azide inhibitor. PMID- 11494224 TI - Riboflavin carrier protein: a serum and tissue marker for breast carcinoma. AB - We have earlier shown that the estrogen-modulated riboflavin carrier protein (RCP) first isolated from the chicken egg is evolutionarily conserved in mammals and is elaborated by lactating mammary gland as demonstrated with rat mammary epithelial cells in culture and confirmed by isolation of the vitamin carrier from bovine milk. In view of several earlier reports that many milk proteins as well as other estrogen-inducible proteins are up-regulated and secreted into circulation in animal models and in women with neoplastic breast disease, we analyzed serum RCP levels in a double-blind study using a specific radioimmunoassay in pre- and post-menopausal women with clinically diagnosed breast cancer at early and advanced stages of the disease and compared these levels with those in normal age-matched control volunteers. Our data reveal that the serum RCP levels in cycling breast cancer patients are 3- to 4-fold higher (p < 0.01) than those in their normal counterparts. This difference in circulatory RCP levels between cancer patients and their age-matched normal counterparts is further magnified to 9- to 11-fold (p < 0.005) at the post-menopausal stage. In addition, there seems to be a good correlation between rising RCP levels and disease progression, since significantly higher RCP concentrations (p < 0.005) are encountered in patients with advanced metastasizing breast cancer versus those with early disease. Using specific monoclonal antibodies, RCP could be localized immunohistochemically in the cytoplasm of invading neoplastic cells of lobular and ductal carcinomas of the breast, indicating that the malignant cells are probably the source of the elevated serum RCP levels in breast cancer. These findings suggest that measurement of circulatory RCP and the immunohistochemical staining pattern of RCP in biopsy specimens could be exploited as an additional marker in diagnosis/prognosis of breast cancer in women. PMID- 11494225 TI - Prognostic value of morphometry in patients with normal breast tissue or usual ductal hyperplasia of the breast. AB - Women with usual ductal hyperplasia have a relative risk of 1.6-1.9 of subsequent breast cancer development. This slightly increased risk is generally not considered sufficiently high to justify (chemo)preventive therapy. It is therefore important to identify high-risk ductal hyperplastic lesions that would benefit from such a treatment. Nuclear morphometric features have been shown in previous work to be useful for objectively describing morphologic features associated with high risk in (pre)invasive breast lesions. The aim of this study was to evaluate whether such morphometric features can also predict subsequent invasive cancer development in patients with the common pattern of usual ductal hyperplasia or a normal breast biopsy. The present case-control study included 423 women with normal breast biopsies (n = 89) or biopsies containing usual ductal hyperplasia (n = 334). Of these 423 women, 132 developed invasive breast cancer during follow-up (mean 16.7 +/- 7.0 years). On the original hematoxylin and eosin-stained sections, nuclear morphometry was performed with a digitizing video overlay system, and mitotic and apoptotic indices were assessed. Patients with mean nuclear feature values for area, perimeter, diameter or longest axis above the 75th percentile had 1.6-1.7 times the breast cancer risk of women with mean nuclear feature values below this value. Pairwise combinations of these features yielded slightly higher cancer risks for the fourth quartile patients, with the highest risk (1.9) for patients with SD of nuclear area and perimeter values above the 75th percentile. The number of apoptotic or mitotic cells had no prognostic value for patients with apparently normal tissue or usual ductal hyperplasia. Our results give a first indication that normal breast tissue or usual ductal hyperplasia harbor nuclear morphologic changes that, when assessed by morphometry, may be used to predict breast cancer development. It is worthwhile studying this further in independent groups of patients with long-term follow-up. PMID- 11494226 TI - Polymerase chain reaction for detection of carcinoembryonic antigen-expressing tumor cells on milky spots of the greater omentum in gastric cancer patients: a pilot study. AB - Our recent studies indicate that omental milky spots are frequently involved in the early stage of peritoneal cancer dissemination. We have used carcinoembryonic antigen (CEA)-specific RT-PCR for omental milky spots to predict peritoneal recurrence in gastric cancer patients. CEA mRNA was found to be positive in both 10 peritoneal washes and 16 greater omenta of 30 gastric cancer patients, including all 6 patients who showed positive results for both cytology and RT-PCR of peritoneal wash and omentum. Three of the 6 cases with positive RT-PCR in the greater omentum but not in the peritoneal wash showed recurrence of peritoneal carcinomatosa within 2 years after operation. Micrometastasis on omental milky spots was histologically confirmed in 6 of 30 gastric cancer cases. Non-specific band was detected only in the omentum of 1 case of 15 benign disease (7%), but not in peritoneal washes (0%), probably due to weak expression of CEA in mesothelial cells. Our results show that CEA-specific RT-PCR targeting micro metastases on omental milky spots is more sensitive than targeting the peritoneal wash or conventional cytology, and suggest that this method is useful for the prediction of peritoneal recurrence in gastric cancer patients. PMID- 11494227 TI - Clinicopathologic and prognostic significance of matrilysin expression at the invasive front in human colorectal cancers. AB - Matrix metalloproteinases (MMPs) have been implicated in tumor progression. Matrilysin, one of the matrix metalloproteinases, is frequently overexpressed in gastrointestinal cancers. The aim of our study was to assess the validity of matrilysin as a prognostic marker of colorectal cancers. Matrilysin expression was immunohistochemically analyzed using formalin-fixed, paraffin-embedded specimens from 113 colorectal cancer patients who had undergone curative surgery. The lumenal surface of neoplastic glands in the superficial layer was apically stained, while the cytoplasm of cancer cells at the invasive front was diffusely stained for matrilysin. Sections with immunostaining signals in more than 30% of carcinoma cells at the invasive front, which were observed in 47 (42%) cases, were judged as being positive for matrilysin. Matrilysin positivity was significantly correlated with the depth of invasion, lymph node metastasis, lymphatic invasion, advanced Dukes' stage and poor outcome. Patients with matrilysin-positive cancer had a significantly shorter overall survival time than those with matrilysin-negative cancer. For patients with intermediate invasive tumor (T2 or T3), only matrilysin was a significant prognostic variable for predicting overall survival in multivariate analysis. Matrilysin expression at the invasive front could be an important marker, predicting an unfavorable prognosis after surgical treatment in patients with colorectal cancer. PMID- 11494228 TI - Expression of three extracellular matrix degradative enzymes in bladder cancer. AB - The relationship between expression of extracellular matrix degradative enzymes, angiogenesis and survival of multistage bladder cancer was determined. Expression of 3 extracellular matrix degradative enzymes (metalloproteinase-2, -9 and heparanase) and microvessel formation were examined in 40 resected bladder cancer specimens by immunohistostochemic staining, and then the association of the enzyme expression with angiogenesis and various stages of cancer was investigated. Heparanase protein expression in muscular invasive or lymph-node metastatic cancer was significantly higher than in superficial or nonmetastatic cancer, respectively (69% vs. 8%, p < 0.001, and 80% vs. 40%, p = 0.028, respectively). Interestingly, heparanase was expressed at much higher levels than matrix metalloproteinase-2 and -9. The mean microvessel count in cancers with heparanase expression was significantly higher than that in cancers without heparanase expression (32.3 +/- 18.2 vs. 5.5 +/- 6.1, p = 0.0008). The microvessel formation was not associated with the expression of matrix metalloproteinase-2 and -9. The cancer-specific and overall survival rates of patients with heparanase expression were significantly lower than those of patients without it (p = 0.0001 and p = 0.0008, respectively). Multivariate analysis showed that heparanase expression was a significantly independent prognostic factor for both cancer-specific (p = 0.0047) and overall survival (p = 0.0200). Our study suggested that heparanase plays important roles in invasion, angiogenesis and metastasis of bladder cancer, and thus, this molecule could be a new molecule to inhibit invasion, angiogenesis and metastasis of bladder cancer. Moreover, our results indicate that expression of heparanase could be a new prognostic factor of this disease. PMID- 11494229 TI - Subcellular localisation of cyclin D1 protein in colorectal tumours is associated with p21(WAF1/CIP1) expression and correlates with patient survival. AB - We investigated the expression of the cell cycle regulatory proteins cyclin D1 and p21(WAF1/CIP1) (p21) in human colorectal carcinomas using immunohistochemistry. Cyclin D1 was not detected in normal colonic epithelium; however, expression was observed in 74/126 (58.7%) of the tumour samples studied. Protein was detected in the nucleus in 22/126 (17.4%) and exclusively in the cytoplasm in 52/126 (41.3%) tumours. Nuclear expression of cyclin D1 was associated with poorly differentiated tumours (p = 0.035) and was more common in right- than in left-sided tumours (p = 0.005). Tumours displaying either, expression of cytoplasmic, (p = 0.05, HR 0.56, 95% CI 0.31-1.0) or nuclear (p = 0.021, HR 0.24, 95% CI 0.07-0.81) cyclin D1 were associated with improved patient survival compared with tumours negative for cyclin D1. p21 protein was strongly expressed mainly in the upper crypts of normal colonic epithelial cells, but in 63/126 (50%) of the tumour samples studied p21 expression was absent. Patients with tumours in which >50% of cells expressed p21 had improved survival compared to patients whose tumours were negative or had < or =50% of cells expressing p21 (p = 0.06, HR 0.33, 95% CI 0.1-1.0). We also observed a significant association between cyclin D1 subcellular localisation and p21 expression: 21/22 (95.5%) tumours expressing cyclin D1 in the nucleus also expressed p21, whereas only 17/52 (32.7%) of the tumours displaying exclusive cytoplasmic cyclin D1 staining were positive for p21 (p < 0.001). These data highlight the significance of exclusive cytoplasmic expression of cyclin D1 in colorectal cancer and lend support to recent in vitro studies suggesting that p21 protein may modulate the subcellular localisation of the cyclin D1 protein. Thus, deregulated expression of the cyclin D1 and p21 proteins are important in colorectal tumourigenesis and have implications for patient prognosis. PMID- 11494230 TI - Reverse transcription-polymerase chain reaction assay for multiple mRNA markers in the detection of breast cancer metastases in sentinel lymph nodes. AB - The identification of specific tumor mRNA markers by reverse transcription polymerase chain reaction might be a valuable diagnostic adjunct for the detection of breast cancer metastases in axillary sentinel lymph nodes (SLNs). In this study we have compared the diagnostic accuracy of an extensive histopathologic examination of 146 SLNs from 123 breast carcinoma patients with that of the evaluation of 5 mRNA markers. When analyzed individually, none of the different markers attained a sensitivity higher than 77.8%, and the general concordance with the histopathologic findings ranged from 78.8 to 83.6%. In a multiple-marker assay, taking into account the expression of at least 1 of the 5 tumor markers, the sensitivity of the test rose to 95.6%, with a specificity of 66.3% and a general concordance with the histopathologic status of 75.3%. Finally, when at least 2 of 3 markers (maspin, cytokeratin 19 and mammaglobin 1) were expressed, the concordance with either SLN or axillary lymph node status was highest (88.4% and 84.6%, respectively). The high prevalence of positive reverse transcription-polymerase chain reaction assays in histologically uninvolved SLNs, however, may hamper extensive application of these techniques in the clinical setting. PMID- 11494231 TI - Preoperative serum concentration of hCGbeta as a prognostic factor in ovarian cancer. AB - In spite of a gradual improvement, survival in epithelial ovarian cancer is disappointingly low. New therapeutic regimens are emerging, and it would be important to be able to predict the prognosis and to stratify patients for clinical trials before therapy. We have evaluated the prognostic value of the pretreatment serum concentrations of 3 tumor markers. The free beta subunit of human chorionic gonadotropin (hCGbeta), CA125 and tumor-associated trypsin inhibitor (TATI) were measured in pretreatment serum samples from 146 patients treated for ovarian cancer between 1990-1995. The patients were followed up until 1998. Elevated concentrations of hCGbeta, CA125 and TATI were observed in 29%, 79% and 33%, respectively. When tested as single variables in Cox's proportional hazards model, stage, grade, size of residual tumor and hCGbeta (all p < 0.001) and CA125 (p = 0.004) correlated with prognosis. However, when fitted as multiple variables together with stage, grade and age in the same model, hCGbeta (RR = 3.42) stage (RR = 2.77) and grade (RR = 3.80) were the only significant variables. When serum hCGbeta was normal, 5-year survival was 80%, but it was only 22% when hCGbeta was elevated. In patients with stage III or IV and minimal residual disease, 5-year survival was 75% if hCGbeta was normal compared with 0% if hCGbeta was elevated. hCGbeta in serum is a strong independent prognostic factor in epithelial ovarian cancer, and its prognostic value is similar to that of grade and stage. The availability of this marker before surgery could facilitate selection of treatment modalities. PMID- 11494232 TI - Prognostic significance of BAG-1 expression in nonsmall cell lung cancer. AB - The purpose of this study was to evaluate the expression of BAG-1 in a cohort of patients with nonsmall cell lung cancer (NSCLC). The intensity and subcellular distribution of BAG-1 expression were correlated with overall survival. Tumor samples were collected from 85 patients diagnosed with NSCLC between 1993-1995 in St. John's, Newfoundland. Expression of BAG-1 was determined by immunohistochemistry using polyclonal anti-BAG-1 antibody. There was significant variation in the immunohistochemical staining patterns of BAG-1, including nonstaining and staining of either the cytoplasm, nucleus or both. Univariate Cox regression analysis showed that those patients whose tumor overexpressed BAG-1 had a significant reduction in the risk of death (hazard ratio = 0.53, p = 0.03). The survival advantage of patients with BAG-1 overexpression tumor was also demonstrated by Kaplan-Meier analysis and log-rank tests (median survival 30.10 months versus 17.04 months, p = 0.05). In addition, multivariate Cox regression analysis showed that patients whose tumor exhibited intense cytoplasmic staining had a further reduction of the risk of death (hazard ratio = 0.42, p = 0.03) and this effect was independent of age, stage and histology. All stages were included in the analysis. Our preliminary data strongly indicate that further investigation is warranted to better define the role of BAG-1 as an independent prognostic factor in NSCLC. PMID- 11494233 TI - Clinical and biologic heterogeneity of hereditary nonpolyposis colorectal cancer. AB - MMR gene mutations and MSI are not found in all clinically diagnosed HNPCC families. We evaluated whether MMR genotyping and tumor MSI analysis could identify distinct clinical subgroups among HNPCC families. Twenty-nine clinical HNPCC families were divided into 3 groups: A, families with hMLH1 or hMSH2 gene mutations; B, MMR gene mutations not present but MSI present in at least 50% of tumors tested; C, mutational and MSI analyses negative. We evaluated tumor spectrum, age at onset, risk of cancer in the follow-up and survival for CRC in the 3 groups. Tumors of the target organs in HNPCC (colon and rectum, endometrium, ovary, small bowel, stomach, renal pelvis and ureter) were more frequent in the first 2 groups than in the latter. Colon cancer was more frequently located in the proximal colon and showed an earlier age at onset in families with MMR gene mutation or with MSI than in families with stable tumors. Comparing the occurrence of tumors in the follow-up, in the first 2 groups patients younger than 50 years had a higher RR, which was particularly marked for CRC (RR = 18.6 for group A vs. group C, RR = 16.7 for group B vs. group C). CRC patients in the first 2 groups had a better clinical prognosis. The results of molecular analysis could distinguish, within clinically defined HNPCC families, different subgroups to which specific programs of surveillance could be addressed. PMID- 11494234 TI - Linkage disequilibrium pattern in the L-myc gene in Italian and Japanese non small-cell lung-cancer patients. AB - Italian and Japanese non-small-cell lung-cancer patients were genotyped for an intragenic L-myc EcoRI restriction site polymorphism previously reported to be associated with lung-tumor prognosis in Asian populations but not in Caucasians. Screening of the L-myc sequence in Italian samples allowed identification of 2 additional 3'-UTR SNPs, located 2.3-3.0 kb from the EcoRI polymorphism, but no coding polymorphism was found. No significant association was found between any of the 3 SNPs and lung-tumor prognosis in Italian patients, consistent with the reported difference between Caucasian and Asian populations. Moreover, the newly discovered polymorphisms in the Italian group were not present in Japanese patients. Significant LD between EcoRI and the 2 other SNPs was detected in the Italian population, whereas no significant LD between the 2 3'-UTR markers was detected despite their close proximity (0.7 kb). Thus, the disparate conclusions about the role of L-myc polymorphism in tumor prognosis among different populations may rest in population-specific LD between the functional gene and the L-myc polymorphism. PMID- 11494235 TI - Polymorphisms of 5,10-methylenetetrahydrofolate reductase and risk of gastric cancer in a Chinese population: a case-control study. AB - Low dietary folate intake has been associated with increased risk of gastric cancer. The 5,10-methylenetetrahydrofolate reductase (MTHFR) involved in folate metabolism has 2 variants, C677T and A1298C, that result in decreased MTHFR activity and lower plasma folate levels. Therefore, we hypothesized that these 2 variants play a role in gastric carcinogenesis. We tested this hypothesis in a Chinese population-based case-control study of 187 histopathologically confirmed gastric cancer cases and 166 healthy controls frequency-matched by age (+/-5 years), gender and residential area. The 677TT genotype was associated with increased risk for gastric cancer [adjusted odds ratio (OR) = 1.87, 95% confidence interval (CI) = 1.00-3.48] compared to the 677CC genotype. This association was more pronounced for gastric cardia cancer (adjusted OR = 2.47, 95% CI = 1.14-5.32). However, no evidence was found for risk associated with the MTHFR A1298C polymorphism. Our findings support the hypothesis that MTHFR C677T variants contribute to gastric carcinogenesis, particularly in gastric cardia. Larger studies incorporating dietary folate intake and serum levels are needed to confirm our findings. PMID- 11494236 TI - Expression of the p53 and Maspin protein in primary prostate cancer: correlation with clinical features. AB - The serine protease inhibitor Maspin has been reported to inhibit the invasiveness and motility of prostate cancer tumor cells. Additionally, a p53 dependent regulatory pathway of Maspin in prostate cancer cell lines has been indicated. The first aim of our study was to determine the prognostic value of Maspin protein expression for the recurrence-free survival of patients undergoing radical prostatectomy for the treatment of clinically localized prostate cancer. Secondly, Maspin expression was correlated to p53 protein expression in order to gain additional information on a possible and previously suggested regulatory influence of the wild-type p53 protein on the Maspin protein expression. Tumor specimens obtained from 84 patients undergoing radical prostatectomy for localized prostate cancer were investigated for the expression of the Maspin and p53 protein by an immunohistochemic approach. Maspin protein expression was correlated with further patients' and tumor characteristics such as tumor stage, histologic grading, regional lymph node status, p53 protein expression and recurrence-free survival of the patients following radical prostatectomy. After a median follow-up of 64 months (24-197 months), 23 of 40 patients (58%) with a negative or decreased Maspin expression (group 1) developed local recurrence or systemic tumor progression in contrast to 8 of 44 patients (18%) with a retained expression of the Maspin protein (group 2) (p = 0.02; log-rank test). The median recurrence-free survival following radical prostatectomy was 26 months (12-37 months) for group 1 patients and 41 months (5-134 months) for patients from group 2 (p = 0.04). A positive immunohistochemic staining reaction for the p53 protein was significantly correlated with a decreased expression of the Maspin protein (p = 0.015; Spearman correlation coefficient). Additionally, loss of Maspin protein expression was correlated to higher tumor stages (p = 0.002) and an increasing histologic dedifferentiation (p = 0.03). This is the first study to indicate that Maspin protein possibly functions as a clinically relevant inhibitor of tumor progression, preventing the local invasiveness and further systemic progression of prostate cancer. Our investigation delivers first hints for a p53-dependent regulatory pathway of the Maspin protein in human prostate cancer. PMID- 11494238 TI - The cognitive neuroscience paradigm: a unifying metatheoretical framework for the science and practice of clinical psychology. AB - The emerging discipline of cognitive neuroscience (CN) enjoins the efforts of cognitive psychologists, neuroscientists, computer scientists, clinical neurologists, neurophilosophers, and many others working collaboratively across traditional disciplinary boundaries to elucidate the manner in which the physical operations of the brain give rise to the vast panoply of human mental and behavioral events. The present article describes the foundational tenets of the CN metatheoretical framework and contends that the CN framework is capable of providing a coherent, unifying scientific paradigm for the discipline of clinical psychology. Clinical psychology's adoption of the CN paradigm would facilitate (a) its consilient linkage with the natural sciences, (b) resolution of long standing internecine theoretical schisms, and (c) enhanced understanding and treatment of numerous forms of psychopathology. Nevertheless, psychology's historically influential radical behavioral (RB) perspective is not easily reconciled with the CN paradigm. However, unlike CN, RB (a) is not fully consilient with the natural sciences, (b) fails to articulate the proximal causal mechanisms that mediate environment-behavior relations, and (c) engages in "greedy reductionism" in its disavowal of informational levels of complexity in the patterning of neural activity. The article concludes with a discussion of the possibility of theoretical rapprochement between CN and RB. PMID- 11494239 TI - Clinical science and human behavior. AB - The debate between mentalism/cognitivism and behaviorism is analyzed, and it is concluded that behaviorism is the philosophy more closely associated with psychology as a behavioral science, the cognitive approach being more closely aligned with biological science. Specific objections to mentalistic interpretations of behavioral phenomena are detailed, and examples from clinical psychology are used to show the importance of behavioral approaches in applied domains. It is argued that the relation between behavior theory and clinical psychology is critical to the continued advancement of applied psychology. Behavior analysis is offered as a direct, applied extension of behavior theory as well as a highly practical and effective approach for understanding, explaining, and modifying the factors that contribute to and maintain maladaptive behaviors. PMID- 11494240 TI - On mental events, disciplinary boundaries, and reductionism: a reply to Plaud. AB - Plaud's (2001) radical behavioral manifesto suggests that a psychological science based exclusively upon the study of environment-behavior functional contingencies would yield a discipline unencumbered by mentalism, vaguely delineated disciplinary boundaries, or inappropriate reductionism. In reply, we note that: (a) mental events-e.g., thoughts and feelings-are increasingly accessible to objective investigation, and provide an observable proximal causal mechanism for the environmental selection of behavior; (b) the call for pristine disciplinary boundaries is anachronistic, inasmuch as progress in the natural sciences has engendered disciplinary boundaries that are increasingly porous; (c) cognitive neuroscience facilitates a comprehensive understanding of complex human behavior by mapping out the relationship between such behavior and underlying brain events, thereby engaging in an appropriate form of reductionism (constitutive reductionism) that has become a hallmark of the natural sciences; and (d) ironically, it is radical behaviorism, in its disavowal of the informational level of complexity instantiated in brain events, that engages in inappropriate eliminative reductionism (i.e., reducing neural information to "nothing but" its underlying bring states). PMID- 11494241 TI - Plaud on Ilardi and Feldman. AB - A cultural analysis is given to the premise that clinical psychology is a discipline of theoretical fragmentation. It is argued that the discipline will achieve paradigmatic status as an applied science by reestablishing the link between behavioral theory and clinical applications. PMID- 11494242 TI - Cognitive neuroscience and the progress of psychological science: once more with feeling (and other mental constructs). PMID- 11494243 TI - Plaud's rebuttal to Ilardi and Feldman's response. PMID- 11494244 TI - Toward rapprochement: comments on the role of biological science within cognitive neuroscience and radical behaviorism. PMID- 11494245 TI - Paradigms, promises, and the potential of clinical psychology. AB - Both Plaud's and Ilardi and Feldman's articles call for clinical psychology to redefine itself according to a particular paradigm or "unifying framework." This commentary focuses on the nature of clinical psychology as an applied discipline, whether clinical psychology in fact has an urgent need for a unifying framework, and whether radical behaviorism or cognitive neuroscience could provide such a framework. It is concluded that, as an applied field that draws both theory and method from a number of natural and social sciences, clinical psychology is served best by continued development and appropriation of competing scientific viewpoints rather than by fealty to a single perspective or paradigm. PMID- 11494246 TI - A tale of three blind men on the proper subject matter of clinical science and practice: commentary on Plaud's behaviorism vs. Ilardi and Feldman's cognitive neuroscience. AB - Plaud (J Clin Psychol 57, 1089-1102, 1109-1111, 1119-1120) and Ilardi and Feldman (J Clin Psychol 57, 1067-1088, 1103-1107, 1113-1117, 1121-1124) argue for two very different approaches to clinical science and practice (i.e., behavior analysis and cognitive neuroscience, respectively). We comment on the assets and liabilities of both perspectives as presented and attempt to achieve some semblance of balance between the three protagonists embroiled in this current debate. The vision of clinical science we articulate is more ecumenical and evolutionary, rather than paradigmatic and revolutionary. As we see it, the problem clinical psychology faces is much larger than the authors let on; namely, how best to make clinical science meaningful and relevant to practitioners, consumers, the general public, and the behavioral health-care community. Clinical psychology's immediate internal problem is not pluralism with regard to subject matter, worldview, methodology, or school of thought, but pluralism in clinical psychologists' adherence to a scientific epistemology as the only legitimate form of clinical psychology. On this latter point, we still have a very long way to go. PMID- 11494249 TI - Correlation between electrophysiology and morphology of three groups of neuron in the dorsal commissural nucleus of lumbosacral spinal cord of mature rats studied in vitro. AB - The dorsal commissural nucleus (DCN) in the lumbosacral spinal cord receives afferent inputs from the pelvic organs via pudendal and pelvic nerves. Electrophysiological and morphological properties of neurons in the DCN of L6-S1 were examined using whole-cell recordings with biocytin-filled electrodes in transverse slices of mature rat spinal cord. Neurons were categorized into three groups according to their discharge in response to suprathreshold depolarizing pulses; neurons with tonic (19/42) and phasic (13/42) firing patterns, and neurons (10/42) that fired in bursts arising from a Ca(2+)-dependent hump. The predominantly fusiform somata of neurons labeled during recording (n = 31) had on average 3.1 primary dendrites, 7.5 terminating dendritic branches, 3.1 axon collaterals, and 14.2 axon terminations per neuron. The groups were morphologically distinct on the basis of their dendritic branching patterns. Phasic neurons (n = 10) had the most elaborate dendritic branching and the largest numbers of axon collaterals. All tonic neurons (n = 11) had axons/collaterals projecting to the intermediolateral area but none to the funiculi, suggesting that they function as interneurons in local autonomic reflexes. Many axons/collaterals of all phasic neurons lay within the DCN, suggesting that they integrate segmental and descending inputs. Seven of 10 neurons with Ca(2+)-dependent humps had axons/collaterals extending into one of the funiculi, suggesting that they project intersegmentally or to the brain. Ca(2+) hump neurons also had more axons/collaterals within the DCN and fewer in the intermediolateral area than tonic neurons. This correlation between firing pattern and morphology is an important step toward defining the cellular pathways regulating pelvic function. PMID- 11494248 TI - Structure and functional connections of presynaptic terminals in the vertebrate retina revealed by activity-dependent dyes and confocal microscopy. AB - The fluorescent dyes sulforhodamine 101 (SR 101) and FM1-43 were used as activity dependent dyes (ADDs) to label presynaptic terminals in the retinas of a broad range of animals, including amphibians, mammals, fish, and turtles. The pattern of dye uptake was studied in live retinal preparations by using brightfield, fluorescence, and confocal microscopy. When bath-applied to the retina-eyecup, these dyes were avidly sequestered by the presynaptic terminals of virtually all rods, cones, and bipolar and amacrine cells; ganglion cell dendrites and horizontal cells lacked significant dye accumulation. Other structures stained with these dyes included pigment epithelial cells, cone outer segments, and Muller cell end-feet. Studies of dye uptake in dark- and light-adapted preparations showed significant differences in the dye accumulation pattern in the inner plexiform layer (IPL), suggesting a dynamic, light-modulated control of endocytotic activity. Presynaptic terminals in the IPL could be segregated on the basis of volume: bipolar varicosities in the IPL were typically larger than those of amacrine cells. The combination of retrograde labeling of ganglion cells and presynaptic terminal labeling with ADDs served as the experimental preparation for three-dimensional reconstruction of both structures, based on dual detector, confocal microscopy. Our results demonstrate a new approach for studying synaptic interactions in retinal function. These findings provide new insights into the likely number and position of functional connections from amacrine and bipolar cell terminals onto ganglion cell dendrites. PMID- 11494250 TI - Thalamic terminal morphology and distribution of single corticothalamic axons originating from layers 5 and 6 of the cat motor cortex. AB - We investigated the axonal morphology of single corticothalamic (CT) neurons of the motor cortex (Mx) in the cat thalamus, using a neuronal tracer, biotinylated dextran amine (BDA). After localized injection of BDA into the Mx, labeled CT axons were found ipsilaterally in the thalamic reticular nucleus (TRN), the ventroanterior-ventrolateral complex (VA-VL), the central lateral nucleus (CL), the central medial nucleus, and the centromedian nucleus, but with the primary focus in the VA-VL. The terminals in the VA-VL formed a large laminar cluster, which extended approximately in parallel with the internal medullary lamina. The laminar organization mirrored morphologic features of single CT axons. We reconstructed the trajectories of 25 single CT axons that arose from layer V (16 axons) or layer VI (9 axons) and terminated in the VA-VL. Terminals of single CT axons that originated from both layer V and layer VI were confined within a laminar structure about 700 microm thick, suggesting the existence of laminar input organization in the VA-VL. Otherwise, the two groups of the CT axons showed contrasting features. All of the CT axons derived from layer VI gave rise to a few short collaterals to the TRN and then formed extensive arborization with numerous small, drumstick-like terminals in the VA-VL. On the other hand, the CT axons arising from layer V gave rise to collaterals whose main axons descended into the cerebral peduncle. Each collateral projected to the VA-VL or CL without projection to the TRN and formed a few small clusters of giant terminals. The two groups of CT neurons in the same cortical column had convergent rather than segregated termination in the VA-VL. However, the terminals of layer VI CT neurons were distributed diffusely and widely in the VA-VL, whereas the terminals of layer V CT neurons were much more focused and surrounded by the terminals of the former group. These contrasting features of the two types of CT projections appear to represent their different functional roles in the generation of motor commands and control of movements in the Mx. PMID- 11494251 TI - Expression of the intermediate filament protein nestin by sustentacular cells in mature olfactory neuroepithelium. AB - The intermediate filament protein nestin has been widely used as a marker for proliferating neural progenitor cells in the nervous system. The mammalian olfactory neuroepithelium is a region of the nervous system that robustly supports ongoing neurogenesis, yet where nestin has not been reported to mark proliferating progenitors. Using immunohistochemistry, we examined nestin expression in the mature olfactory neuroepithelium and found it to be tightly restricted to the basal compartment where the olfactory neuronal progenitor cell population resides. The pattern of nestin immunoreactivity was consistent with expression by the endfeet and inferior processes of sustentacular cells rather than basal cells. Using a bank of defined antibody markers, we confirmed nestin's pattern of distribution to be different from that of cytokeratin, vimentin, GBC 1, GAP43, and carnosine. It was highly similar to the pattern of SUS-4 immunoreactivity in the basal region of the neuroepithelium. Following surgical bulbectomy, nestin expression was up-regulated and became evident in the cell bodies of sustentacular cells situated more apically in the neuroepithelium. We have shown nestin to be present in the basal region of the adult olfactory neuroepithelium in the zone that supports ongoing neurogenesis in the adult, but its expression is restricted to the inferior parts of sustentacular cells rather than the neuronal progenitor cells. Nestin may play a potential role in the migration of recently proliferated olfactory neurons on the scaffolding of sustentacular cells in a manner analogous to its proposed role in radial glia during embryonic development of the central nervous system. PMID- 11494252 TI - Localization of two high-threshold potassium channel subunits in the rat central auditory system. AB - The firing pattern of auditory neurons is determined in part by the type of voltage-sensitive potassium channels expressed. The expression patterns for two high-threshold potassium channels, Kv3.1 and Kv3.3, that differ in inactivation properties were examined in the rat auditory system. The positive activation voltage and rapid deactivation kinetics of these channels provide rapid repolarization of action potentials with little effect on action potential threshold. In situ hybridization experiments showed that Kv3.3 mRNA was highly expressed in most auditory neurons in the rat brainstem, whereas Kv3.1 was expressed in a more limited population of auditory neurons. Notably, Kv3.1 mRNA was not expressed in neurons of the medial and lateral superior olive and a subpopulation of neurons in the ventral nucleus of the lateral lemniscus. These results suggest that Kv3.3 channels may be the dominant Kv3 subfamily member expressed in brainstem auditory neurons and that, in some auditory neurons, Kv3.1 and Kv3.3 may coassemble to form functional channels. The localization of Kv3.1 protein was examined immunohistochemically. The distribution of stained somata and neuropil varied across auditory nuclei and correlated with the distribution of Kv3.1 mRNA-expressing neurons and their terminal arborizations, respectively. The intensity of Kv3.1 immunoreactivity varied across the tonotopic map in the medial nucleus of the trapezoid body with neurons responding best to high frequency tones most intensely labeled. Thus, auditory neurons may vary the types and amount of K(+) channel expression in response to synaptic input to subtly tune their firing properties. PMID- 11494253 TI - Bipolar cell diversity in the primate retina: morphologic and immunocytochemical analysis of a new world monkey, the marmoset Callithrix jacchus. AB - The aim of this study was to identify the bipolar cell types in the retina of a New World monkey, the common marmoset, and compare them with those found in the Old World macaque monkey. Retinal whole-mounts, sections, or both, were stained by using DiI labeling and immunohistochemical methods. Semithin sections were analyzed by using quantitative methods. We show that the same morphologic types of bipolar cell as described for the Old World macaque monkey by Boycott and Wassle (Boycott and Wassle [1991] Eur. J. Neurosci. 3:1069-1088) are present in marmoset retina: two types of midget bipolar cells, six type of diffuse bipolar cells, a blue cone bipolar cell, and one type of rod bipolar cell. The pattern of staining with different immunohistochemical markers ("fingerprint") of each bipolar cell type in marmoset was also the same as described for macaque, with one exception: the flat midget bipolar cell (FMB) class is labeled by antibodies to recoverin in macaque but is labeled by antibodies to CD15 in marmoset. The labeled FMB cells in marmoset make contact with multiple cone photoreceptors throughout most of the extrafoveal retina. The spatial density of bipolar cells in marmoset is shown to be sufficient to support one-to-one connectivity of midget bipolar and ganglion cells in the fovea and to allow for parallel pathways to ganglion cells throughout the retina. Quantitative differences in the morphology and receptor connectivity between marmoset and macaque can be related to differences in cone and rod photoreceptor density between the species. We conclude that bipolar cell diversity is a preserved feature of the primate retina. PMID- 11494254 TI - Immunohistochemical localization of atrial natriuretic factor and autoradiographic distribution of atrial natriuretic factor-binding sites in the brain of the cave salamander Hydromantes genei (Amphibia, Plethodontidae). AB - The distribution of atrial natriuretic factor (ANF)-like immunoreactivity in the central nervous system of the cave salamander Hydromantes genei (Amphibia, Plethodontidae) was investigated by using antisera raised against rat and human ANF(1-28). Concurrently, the location of ANF-binding sites was determined by autoradiography, using radioiodinated human ANF(1-28) as a tracer. In several regions of the brain, including the olfactory bulb, the preoptic area, the ventral thalamus, the tectum of the mesencephalon, and the choroid plexuses inside the ventricles, a good correlation was observed between the distribution of ANF-immunoreactive elements and the location of ANF-binding sites. Mismatching was found in the habenular nucleus, the commissura habenularis, the fasciculum retroflexus, and the interpeduncular nucleus, which contained high levels of binding sites but were devoid of ANF-immunoreactive structures. In contrast, a few other regions, such as the pineal gland and the subcommissural organ, showed a high concentration of ANF-like immunoreactivity but did not contain ANF-binding sites. This study provides the first localization of ANF-like immunoreactivity and ANF-binding sites in the brain of an urodele amphibian. The results show that the ANF peptidergic system in the cave salamander has an organization more simple than the organizations described for the brain of frog or other vertebrates. This feature is probably related to the expression of highly pedomorphic characters in plethodontids. The anatomical distribution of ANF-immunoreactive elements and ANF binding sites suggests that ANF-related peptides may act as hypophysiotropic hormones as well as neurotransmitters and/or neuromodulators in the salamander brain. PMID- 11494255 TI - Axonal topography of cortical basket cells in relation to orientation, direction, and ocular dominance maps. AB - The axonal (bouton) distributions of a layer 4 clutch cell (CC), two layer 3 medium-sized basket cells (MBC), and a layer 3 large basket cell (LBC) to orientation, direction, and ocular dominance maps were studied quantitatively. 1) The CC provided exclusively local projections (<380 microm from the soma) and contacted a narrow "niche" of functional representations. 2) The two MBCs emitted local projections (75% and 79% of all boutons), which were engaged with isoorientations (61% and 48%) and isodirections, and long-range projections (25% and 21%, >313 microm and >418 microm), which encountered cross-orientation sites (14% and 12%) and isoorientation sites (7% and 5%). Their direction preferences were mainly perpendicular to or opposite those of local projections. 3) The LBC provided the majority (60%) of its boutons to long-range distances (>437 microm). Locally, LBC boutons showed a rather balanced contribution to isoorientations (19%) and cross-orientations (12%) and preferred isodirections. Remotely, however, cross-orientation sites were preferred (31% vs. 23%) and the directional output was balanced. 4) Monte Carlo simulations revealed that the differences between the orientation specificity of local and long-range projections cannot be explained by a homogeneous lateral distribution of the boutons. 5) There was a similar eye preference in the local and long-range projection fields of the MBCs. The LBC contacted both contra- and ipsilateral eye domains. 6) The basket axons showed little laminar difference in orientation and direction topography. The results suggest that an individual basket cell can mediate a wide range of effects depending on the size and termination pattern of the axonal field. PMID- 11494256 TI - Co-expression of estrogen and thyroid hormone receptors in individual hypothalamic neurons. AB - Estrogen receptors (ER) and thyroid hormone receptors (TR) are members of the nuclear receptor family of transcription factors that induce or repress the expression of target genes. Previous behavioral studies in female rodents have demonstrated that thyroid hormones can antagonize the effects of estrogen in the central nervous system (CNS), particularly by attenuating estrogen's ability to facilitate reproductive behaviors. Additional molecular studies have suggested a mechanism for this antagonism by showing that ligand-activated ER alpha and TRs have the potential to interact in their transcriptional controls. Although the expression patterns of ER alpha and TRs in the rodent brain appear to overlap in behaviorally relevant areas, it remained to be determined whether these two classes of proteins coexist in vivo at the level of single neurons. To address this possibility, we employed a highly sensitive double-label in situ hybridization technique using digoxigenin and (35)S-labeled cRNA probes to analyze, in detail, the expression of ER alpha mRNA with TR alpha 1 and TR alpha 2 mRNAs in the same neurons of the ovariectomized (OVX) adult mouse brain. Our results demonstrate that a large majority of the ER alpha-positive neurons also expresses TR alpha 1 and TR alpha 2 mRNAs. Quantitative examination of the cellular expression in the ventromedial and arcuate nuclei of the hypothalamus (VMH and Arc) showed that 81.5% and 80.5% of the neurons endowed with ER alpha mRNA also contain TR alpha 1 and TR alpha 2 mRNAs, respectively. In the amygdala, more than 60.5% and 67% of ER alpha-positive cells also contain TR alpha 1 and TR alpha 2 mRNAs, respectively. These findings provide the first anatomical evidence that ER and TR can be found in the same neurons, including hypothalamic neurons. This coexpression of ER alpha and TR provides the cellular basis for a new level of neuronal integration in a brain region where estrogens control female reproductive behaviors. PMID- 11494257 TI - Down-regulation of trkA mRNA within nucleus basalis neurons in individuals with mild cognitive impairment and Alzheimer's disease. AB - Recent studies indicate that trkA expression is reduced in end-stage Alzheimer's disease (AD). However, understanding the neuropathologic correlates of early cognitive decline, as well as the changes that underlie the transition from nondemented mild cognitive impairment (MCI) to AD, are more critical neurobiological challenges. In these regards, the present study examined the expression of trkA mRNA in individuals diagnosed with MCI and AD from a cohort of people enrolled in a Religious Orders Study. Individuals with MCI and AD displayed significant reductions in trkA mRNA relative to aged-matched controls, indicating that alterations in trkA gene expression occur early in the disease process. The magnitude of change was similar in MCI and AD cases, suggesting that further loss of trkA mRNA is not necessarily associated with the transition of individuals from nondemented MCI to AD. The loss of trkA mRNA was not associated with education, apolipoprotein E allele status, gender, Braak score, global cognitive score or Mini-Mental Status Examination. In contrast, the loss of trkA mRNA in MCI and AD was significantly correlated with function on a variety of episodic memory tests. PMID- 11494258 TI - Development of three distinct GnRH neuron populations expressing two different GnRH forms in the brain of the African catfish (Clarias gariepinus). AB - The early development of both the catfish gonadotropin-releasing hormone (cfGnRH) and the chicken GnRH-II (cGnRH-II) system was investigated in African catfish by immunocytochemistry by using antibodies against the GnRH-associated peptide (GAP) of the respective preprohormones. Weakly cfGnRH-immunoreactive (ir) neurons and fibers were present at 2 weeks after hatching (ph) but only in the ventral telencephalon and pituitary. Two weeks later, cfGnRH fibers and neurons were also observed in more rostral and in more caudal brain areas, mainly in the preoptic area and hypothalamus. Based on differences in temporal, spatial, and morphologic appearance, two distinct cfGnRH populations were identified in the ventral forebrain: a population innervating the pituitary (ventral forebrain system) and a so-called terminal nerve (TN) population. DiI tracing studies revealed that the TN population has no neuronal connections with the pituitary. The cGnRH-II system is present from 2 weeks ph onward in the midbrain tegmentum and only their size and staining intensity increased during development. Based on the comparison of GnRH systems amongst vertebrates, we hypothesize that during fish evolution, three different GnRH systems evolved, each expressing their own molecular form: the cGnRH-II system in the midbrain, a hypophysiotropic GnRH system in the hypothalamus with a species-specific GnRH form, and a salmon GnRH-expressing TN population. This hypothesis is supported by phylogenetic analysis of known GnRH precursor amino acid sequences. We hypothesize, because the African catfish is a less advanced teleost species, that it contains the cfGnRH form both in the ventral forebrain system and in the TN population. PMID- 11494259 TI - Neuropeptide Y immunoreactivity in cutaneous sympathetic and sensory neurons during development of the guinea pig. AB - Different levels of the cutaneous vasculature are innervated selectively by subpopulations of sympathetic neurons distinguished by the presence or absence of immunoreactivity (-IR) for neuropeptide Y (NPY). This study used multiple labelling immunohistochemistry to examine the appearance of NPY-IR in neurons innervating cutaneous vessels in the ear pinna of embryonic, fetal, and neonatal guinea pigs. NPY-immunoreactive axons were detected in the ear bud at embryonic day 25. However, these axons lacked IR for tyrosine hydroxylase (TH) and often ran in bundles with substance P (SP)-immunoreactive axons close to the epidermis. Many neuronal somata in the cervical dorsal root ganglia (DRG) at late embryonic stages contained NPY-IR with or without SP-IR, but no NPY-IR was detected in DRG or subepidermal axons by late fetal stages. IR for calcitonin gene-related peptide increased in DRG neurons from midfetal to late fetal stages, after the decrease in NPY-IR. Populations of TH-IR neurons with or without NPY-IR were present in the superior cervical ganglion (SCG) from midembryonic stages. TH immunoreactive axons were not detected in the ear pinna until midfetal stages, when axons with TH-IR and NPY-IR innervated proximal arteries and TH immunoreactive axons without NPY-IR innervated distal vessels. Vasoactive intestinal peptide-IR was detected transiently in most fetal SCG neurons with TH IR and NPY-IR but was not detected in cutaneous axons. These results demonstrate that selective expression of NPY by subpopulations of sympathetic neurons occurs prior to innervation of their targets. This suggests that target contact is not required to establish appropriate patterns of expression of peptide neurotransmitters by cutaneous sympathetic neurons. PMID- 11494260 TI - GABA-immunoreactive neurons in the mushroom bodies of the honeybee: an electron microscopic study. AB - Synaptic contacts of gamma-aminobutyric acid (GABA) -immunoreactive neurons in honeybee mushroom bodies were studied by using electron microscopic immunocytochemistry. In the lip region of the calyx neuropil, GABA-immunoreactive profiles formed synapses onto both small postsynaptic profiles (76%) and large immunonegative boutons (4%), which were likely to belong to the intrinsic and extrinsic mushroom body neurons, respectively. Three morphologic types of the large immunonegative boutons were distinguished: "light," "dark," and "dense core"; all of them received synaptic inputs from the GABA-immunoreactive profiles. A significant proportion of the synapses formed by the GABA immunoreactive neurons in the lip region (20%) were input synapses from immunonegative neurons. Analysis of thin serial sections showed that the output and input synapses formed microcircuits in which both large immunonegative boutons and small postsynaptic profiles were involved. We interpret these findings to show that negative feedforward and feedback loops exist within the microcircuits of the lip region. PMID- 11494261 TI - Spatiotemporal pattern of nicotinamide adenine dinucleotide phosphate-diaphorase reactivity in the developing central nervous system of premetamorphic Xenopus laevis tadpoles. AB - We have catalogued the progressive appearance of putative nitric oxide synthase (NOS)-containing neurons in the developing central nervous system (CNS) of Xenopus laevis. Xenopus embryos and larvae were processed in wholemount and in cross section using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH d) histochemistry as a marker for NOS within the CNS. The temporal sequence of NADPH-d reactivity identified discrete groups and subgroups of neurons in the forebrain, midbrain, and hindbrain on the basis of their morphology, location, and order of appearance during development. A proportion of these groups of neurons appeared to be important in sensory processing and motor control. Staining also appeared at specific stages in the spinal cord, the retina, and the skin. After the appearance of labelling, NADPH-d reactivity continued in each of the cell groups throughout the stages examined. We found no evidence for staining that subsequently disappeared at later stages in any cell group, indicating a persistent rather than transient role for NO in the Xenopus tadpole CNS. These results are discussed in light of recent findings on possible roles for NADPH-d positive cell groups within the developing motor circuitry. PMID- 11494262 TI - Structure and response patterns of olfactory interneurons in the honeybee, Apis mellifera. AB - To analyze morphologic and physiological properties of olfactory interneurons in the honeybee, Apis mellifera, antennal lobe (AL) neurons were intracellularly recorded and subsequently labeled with Neurobiotin. Additional focal injections were carried out with cobalt hexamine chloride and dextran fluorescent markers. Olfactory interneurons (projection neurons, PNs) project by means of five tracts, the lateral, the median, and three mediolateral antennocerebral tracts (l-, m-, and ml-ACT, respectively) to the mushroom bodies (MBs) and the protocerebral lobe (PL) of the ipsilateral protocerebrum. Uniglomerular PNs of the m- and l-ACT receiving input from a single glomerulus of the AL also arborize in different regions of the AL. The vast majority of l-ACT innervate the T1 region, whereas m ACT neurons arborize exclusively in the T2, T3, and T4 regions (T1-4 : AL projection area of sensory cells from the antennae). In the calyces of the MB, uniglomerular PNs form varicosities in the basal ring and the lip region. Individual neurons of both types exhibit unequal innervation within and between the two calyces. In addition, m-ACT fibers ramify more densely within the lip neuropil and show a higher incidence of spine-like processes than l-ACTs. In the PL, l-ACTs arborize exclusively within the lateral horn, whereas some m-ACT neurons innervate a broader region. Multiglomerular neurons of the ml-ACT leave the AL by means of three subtracts (ml-ACT 1-3). Two different types can be distinguished according to their protocerebral target areas: ml-ACTs projecting to the lateral PL (LPL) and to the neuropil around the alpha-lobe (tracts 2 and 3) and neurons projecting only to the LPL (tract 1). Intracellular recordings indicate that both l- and m-ACT neurons respond to general odors but with different response properties, indicating that odor information is processed in parallel pathways with different functional characteristics. Just like m-ACT neurons, ml-ACT neurons respond to odors with complex activity patterns. Bilateral interneurons, originating in the suboesophageal ganglion, connect glomeruli of both AL, and send an axon through the m-ACT in each hemisphere of the brain, terminating in the lip region of the calyces. These neurons respond to contact chemical stimuli. PMID- 11494263 TI - Impaired glucose tolerance--does it cause neuropathy? PMID- 11494264 TI - Contractile properties of human motor units in health, aging, and disease. AB - The primary function of skeletal muscle is to produce force for postural control and movement. Although the contractile properties of the whole muscle are useful functional indicators, they do not accurately reflect the heterogeneity of the constituent motor units (MUs) and their changes in health and disease. However, data on the contractile properties of human MUs, in comparison to other animal species, are relatively sparse. This, in part, is due to greater methodological challenges of in vivo studies of MUs in the human. The purpose of this review is to critically appraise the methods used in humans; to describe the normative data from different muscle groups; to discuss differences between data from healthy humans and other animal species; and, last, to characterize changes of the MU contractile properties in aging, disease, and in response to intervention. Because the spike-triggered averaging technique can only be used to study the twitch properties, other methods were subsequently developed to measure a wider range of contractile properties. Although there is general agreement between human data and those from other animal species, major differences do exist. Potential reasons for these discrepancies include true biological differences, but differences in the techniques used may also be responsible. Although limited, measurement of MU contractile properties in humans has provided insight into the changes associated with aging and motoneuronal diseases and provides a means of gauging their adaptive capacity for training and immobilization. PMID- 11494265 TI - Influence of age and height on nerve conduction. AB - Previous studies demonstrated age- and height-related slowing of nerve conduction velocity (NCV) and reduction in nerve response amplitude. Unfortunately, many studies examined discontinuous populations, preventing regression analysis. The correlation coefficients of available studies vary widely, preventing clear conclusions. We retrospectively examined 3969 clinically normal subjects ranging in age from 20 to 95 years from a total of 22,420 electrodiagnostic studies done between 1986 and 1998. The correlation of NCV with height was stronger than with age. Regression equations using both factors account for 12--27% of the variance. Responses were seen in the majority of patients aged 70 years and older, but the percentage of normals who had no response increased with advancing age. Age was strongly inversely correlated with the amplitudes of both sensory and motor responses, accounting for 7--16% of the variance. Regression equations using both height and age improved this correlation, accounting for 7--22% of the variance. Therefore, both height and age must be taken into consideration when normal values are developed. PMID- 11494266 TI - Modulation of H reflex of pretibial and soleus muscles during mastication in humans. AB - A previous study in our laboratory demonstrated that the soleus H reflex was facilitated during mastication in humans. In the present study, we investigated whether there was any modulation of the magnitude of the pretibial H reflex during mastication in five healthy adult volunteers. The pretibial H reflex was significantly facilitated during mastication, and there was no significant difference in the facilitation between jaw-closing and jaw-opening phases; that is, the gain of the H reflex was modulated tonically but not in a phase-dependent manner during mastication. Furthermore, in the same subjects, we confirmed that the soleus H reflex was facilitated during mastication. Based on our findings, we conclude that the H reflexes in both the pretibial and soleus muscles undergo a nonreciprocal facilitation during mastication. It is suggested that mastication contributes to stabilization of postural stance in humans. PMID- 11494267 TI - Diagnostic strategy for familial and sporadic cases of neuropathy associated with 17p11.2 deletion. AB - Clinical, electrophysiologic and molecular studies were performed on at-risk members of 14 families with hereditary neuropathy with liability to pressure palsies (HNPP), in order to detect asymptomatic carriers of the 17p11.2 deletion. Sporadic cases due to de novo deletion accounted for 21% of the investigated HNPP families. Approximately one half of deletion carriers were asymptomatic and did not display significant signs on clinical examination. The electrophysiologic hallmark in both symptomatic and asymptomatic deletion carriers was the presence of a nonuniform sensorimotor demyelinating polyneuropathy with conduction abnormalities preferentially located at common entrapment sites and distal nerve segments. A perfect correlation was found between the molecular and electrophysiologic analyses. A reliable screening method to detect clinically unaffected carriers of the deletion in families with HNPP was the evaluation of motor conduction in at least two nerves across usual entrapment sites, especially the ulnar nerve at the elbow, and evaluation of sensory conduction in the sural nerve. In sporadic cases due to a de novo deletion, electrophysiologic studies were suggestive but not sufficient for the diagnosis, and molecular analysis represented the most sensitive diagnostic tool. PMID- 11494268 TI - Incomplete recovery of voluntary isometric force after fatigue is not affected by old age. AB - The 60-min recovery profiles of voluntary and electrically stimulated force, contractile speed, surface electromyography, muscle activation via twitch interpolation, and muscle compound action potentials (M-waves) in the elbow flexors of seven young men (24 +/- 2 years) and seven men over 80 years of age (84 +/- 2 years) were compared following intermittent (3 s on, 2 s off) contractions at 60% of each subject's maximum voluntary contraction (MVC) force. There was no age-related difference between groups in the average time to fatigue or the rate of voluntary force loss; both groups lost 40% of their force within approximately 5 min. Despite a rapid increase to approximately 83% of the prefatigue MVC by the third minute of recovery for both groups, MVC force did not return to the prefatigue value within 60 min (94 +/- 4% young, 91 +/- 3% old). These results suggest that the incomplete recovery of voluntary force was likely due to a peripheral limitation in the muscle at the level of excitation- contraction coupling, and was not affected by age. Delayed recovery of voluntary force and a greater degree of low-frequency fatigue in the old men were not observed and there were no age-related impairments in any parameter normalized to the prefatigue value during fatigue or recovery. We suggest that the specific fatigue task may be more important to recovery than proposed alterations in the aged neuromuscular system when normalization and matching of the fatigue task criteria occurs. PMID- 11494269 TI - Depletion and sizes of motor units in spinal muscular atrophy. AB - Motor unit number estimation (MUNE) was applied to the biceps brachii muscles of 13 young patients (age 5--24 years) with spinal muscular atrophy (SMA) and the results compared with those of healthy control subjects matched for age and gender. In the SMA patients, all motor unit (MU) estimates fell below the control range, and there was good correspondence between the values for the two arms in the same subject. No correlation could be found between the MUNEs and the severity of the weakness. This unexpected result was attributed to the presence of small and normal-sized MUs in the muscles of patients, in addition to MUs that appeared to be considerably enlarged. The threefold mean increase in MU potential size was insufficient to compensate for the MU loss. In addition, the study confirmed that there are, on average, approximately 130 MUs in the healthy biceps brachii muscle. PMID- 11494270 TI - Contractile speed and fatigue of adductor pollicis muscle in multiple sclerosis. AB - The purpose of the study was to investigate differences in contractile speed, force, and fatigability of the adductor pollicis muscle between 12 patients with multiple sclerosis (MS) and 8 sedentary control subjects matched for age and gender. There were no differences between the patients with MS and control subjects with respect to the percentage of maximal muscle force that could be recruited during voluntary effort (95.5 +/- 3.9% and 98.2 +/- 2.0%, respectively, P = 0.10), the stimulation frequency/force and force/velocity relationships, the rates of force development and relaxation, fatigue resistance, and the recovery rate of adductor pollicis muscle. However, previous results from the same group of MS patients showed that quadriceps femoris muscle force and resistance to fatigue were reduced. Therefore, our data support the clinical experience that, in patients with MS, lower limb muscle function is more or earlier affected than upper limb muscle function. PMID- 11494271 TI - Response to high-intensity eccentric muscle contractions in persons with myopathic disease. AB - Although the response to intense eccentric muscle contractions is well described in normal subjects, concern exists about possible untoward effects in persons with myopathic diseases. We investigated 14 subjects with slowly progressive muscular dystrophies including myotonic muscular dystrophy (n = 9), facioscapulohumeral dystrophy (n = 2), limb-girdle syndrome (n = 2), and Becker muscular dystrophy (n = 1). Control subjects consisted of 18 able-bodied persons. Subjects performed two sets of eight maximal-effort eccentric repetitions of the elbow flexors, with measurement of maximal concentric strength, serum creatine kinase, resting and flexed arm angle, arm circumference, and soreness at days 0, 3, and 7. Although the myopathic group had less initial strength, both groups demonstrated a similar response to the protocol over 7 days. Both groups had a significant rise in serum creatine kinase, which was still elevated at 7 days (P < 0.05). The control group demonstrated a slightly greater injury response in terms of soreness, resting and flexed arm angles, and arm swelling. Both groups of subjects appeared to respond similarly to an acute bout of eccentric contractions. However, the potential long-term effects of this type of exercise in persons with myopathic diseases remains unknown. PMID- 11494272 TI - Changes in glycolytic network and mitochondrial design in creatine kinase deficient muscles. AB - Skeletal muscles respond with high plasticity to pathobiological conditions or changes in physiological demand by remodeling cytoarchitectural and metabolic characteristics of individual myocytes. We have previously shown that muscles of mice without mitochondrial and/or cytosolic creatine kinases (ScCKmit(-/-) and/or M-CK(-/-)) partly compensate for the defect(s) by redirecting metabolic pathways and ultrastructural characteristics. Here, we show by semiquantitative Western blot analysis that the compensatory changes involve mutation- and fiber-type specific coordinated regulation of divergent but functionally coupled groups of proteins. Fast-twitch gastrocnemius muscle of CK(--/--) mice display a two- to fourfold upregulation of mitochondrial cytochrome c oxidase, inorganic phosphate carrier, adenine nucleotide translocator, and voltage-dependent anion channel proteins. In parallel, cytosolic myoglobin is upregulated. Slow-twitch soleus muscle responds with changes in the glycolytic enzyme pattern, including a shift in lactate dehydrogenase isoenzyme composition. Adaptations in the network for oxidative adenosine triphosphate (ATP) production are already apparent at 17 days of age. PMID- 11494273 TI - Prospective follow-up study of chronic polyneuropathy of undetermined cause. AB - Forty consecutive patients with chronic sensorimotor polyneuropathy of undetermined cause were followed to determine disease progression and prognosis. They had had neurological symptoms for at least 1 year before presentation. Neurophysiological examination in all patients showed chronic axonal degeneration, which was confirmed by sural nerve biopsy. Patients were reviewed every 3 months. Laboratory tests were performed every 6 months for the first 2 years and then annually. Neurophysiological examination was performed annually in all patients. Patients were followed up for at least 4 years. In no instance was a possible etiological factor detected during follow-up. The clinical and electrophysiological findings had a slowly progressive course. We suggest that patients with a chronic polyneuropathy of undetermined cause despite detailed investigations do not require further extensive and expensive laboratory tests and neurophysiological studies during follow-up. PMID- 11494274 TI - Changes in a rat facial muscle after facial nerve injury and repair. AB - This study describes changes in a rat facial muscle innervated by the mandibular and buccal facial nerve branches 4 months after nerve injury and repair. The following groups were studied: (A) normal controls; (B) spontaneous reinnervation by collateral or terminal sprouting; (C) reinnervation after surgical repair of the mandibular branch; and (D) chronic denervation. The normal muscle contained 1200 exclusively fast fibers, mainly myosin heavy chain (MyHC) IIB fibers. In group B, fiber number and fiber type proportions were normal. In group C, fiber number was subnormal. Diameters and proportions of MyHC IIA and hybrid fibers were above normal. The proportion of MyHC IIB fibers was subnormal. Immediate and delayed repair gave similar results with respect to the parameters examined. Group D rats underwent severe atrophic and degenerative changes. Hybrid fibers prevailed. These data suggest that spontaneous regeneration of the rat facial nerve is superior to regeneration after surgical repair and that immediacy does not give better results than moderate delay with respect to surgical repair. Long delays are shown to be detrimental. PMID- 11494275 TI - Sensory potentials evoked by tactile stimulation of different indentation velocities at the finger and palm. AB - Previous studies suggest that the rate of indentation of a tactile probe determines which skin mechanoreceptors are activated. To further investigate this possibility, indentations of 300 microm at velocities of 100 (T100) and 400 microm/ms (T400) were applied to the tip (FT) and the proximal phalanx of digit III (PP) and the thenar eminence (Pm) of ten healthy volunteers, and compared with responses after electrical stimulation at the FT. Compound sensory action potentials (CSAPs) were recorded from the median nerve through needle electrodes at the wrist and elbow. The maximal sensory conduction velocities (SNCVs) between wrist and elbow were similar with electrical and T400 stimulation, but on average were 15% lower with T100 stimulation (P < 0.001). With both indentation velocities, SNCVs were similar regardless of stimulation sites. Amplitudes of tactile CSAPs with FT stimulation were 1--2 microV at T400 and 0.3--0.4 microV at T100. The CSAP areas evoked by T100 stimulation showed a reduction from fingertip to proximal finger to palm (P < 0.05-0.005), whereas those obtained with T400 stimulation showed a reduction only at the palm (P < 0.05). The results support previous studies indicating that fast indentation at 400 microm/ms activated deeply placed Pacinian corpuscles as well as superficially situated Meissner corpuscles, whereas slower indentation at 100 microm/ms activated primarily Meissner corpuscles. PMID- 11494276 TI - Recovery from fatigue in fast and slow single intact skeletal muscle fibers from aging mouse. AB - In the present work, we studied the recovery from fatigue (RF) of single intact fast- and slow-twitch muscle fibers from young (age 5--7 months) and old (age 22- 24 months) mice. To examine whether differences in RF underlie decreases in muscle strength and endurance with aging, we performed in vitro experiments in manually dissected extensor digitorum longus (EDL) and soleus muscle fibers. We measured the recovery of the maximum force every 5 min for a total period of 30 min after inducing fiber fatigue. Fibers were classified, according to the fatigue index, into the following three groups: 0.75--0.99, 0.5--0.74, and <0.5. Although the tetanic tension of EDL and soleus fibers from young and old mice recovered significantly, no statistically significant difference in tension or recovery time was observed between age groups. These data support the concept that the reported decline in muscle force and endurance with aging is not related to changes in RF of individual muscles fibers. PMID- 11494277 TI - Painful sensory polyneuropathy associated with impaired glucose tolerance. AB - We examined records of 121 patients coded as idiopathic polyneuropathy, extracting neuropathy symptoms, electromyographic data, and diagnostic blood work. Of 89 patients screened for glucose handling, 28 demonstrated frank diabetes mellitus. Of the remaining 61 patients, 15 (25%) had impaired glucose tolerance (IGT) by American Diabetes Association criteria (serum glucose 140--200 mg/dl 2 h after a 75-g glucose load). Excluding those with diabetes mellitus, 35% of patients with neuropathic pain had IGT, more than twice the prevalence found in large, unselected population studies. No other common etiology of polyneuropathy was identified. Two-hour oral glucose tolerance test results were often abnormal, whereas fasting glucose or hemoglobin A1c was normal. Bias due to referral pattern, body weight, or genetics might affect the comparison of our polyneuropathy cohort with a broader, population-based control. However, our results corroborate an association between IGT and painful sensory polyneuropathy and link these patients syndromically to the typical painful polyneuropathy of diabetes mellitus. PMID- 11494278 TI - The frequency of undiagnosed diabetes and impaired glucose tolerance in patients with idiopathic sensory neuropathy. AB - The aim of the study was to determine the frequency of undiagnosed abnormal glucose metabolism in patients with idiopathic sensory neuropathy. Patients were separated into two groups depending on presence or absence of painful symptoms, and an oral glucose tolerance test was performed. Of the 48 patients studied, those with painful symptoms had a higher frequency of abnormal glucose metabolism than literature-based controls, whereas patients without painful symptoms showed no difference. Comparison of patients with and without painful symptoms had a P value of 0.02. The results indicate the need to consider undiagnosed abnormal glucose metabolism in patients with idiopathic sensory neuropathy. PMID- 11494279 TI - Latency of compound muscle action potentials of the anal sphincter after magnetic sacral stimulation. AB - The aim of this study was to present the failure rate and normal values for motor latency of the anal sphincter after magnetic sacral stimulation (LMSS) using a modified recording technique. A bipolar sponge electrode was placed in the anal canal for recording. A ground electrode was placed in the rectum to reduce stimulus artifact. Magnetic stimulation was induced through a twin coil energized by a Maglite-r25 generator. Two groups were examined: 14 healthy volunteers and 14 patients with a spinal cord injury (SCI) above the conus. Nine of 56 studies (16%) failed. There were no significant differences in latency between right- and left-sided stimulation or between the healthy group and the SCI patients. As described, LMSS measurements are minimally invasive and have a low failure rate. They may be used to test the integrity of the distal motor pathway in patients with bladder or bowel dysfunction who may benefit from continuous electrical sacral root stimulation. PMID- 11494280 TI - Practice parameter for repetitive nerve stimulation and single fiber EMG evaluation of adults with suspected myasthenia gravis or Lambert-Eaton myasthenic syndrome: summary statement. PMID- 11494281 TI - Literature review of the usefulness of repetitive nerve stimulation and single fiber EMG in the electrodiagnostic evaluation of patients with suspected myasthenia gravis or Lambert-Eaton myasthenic syndrome. AB - A retrospective literature review of the electrodiagnosis of myasthenia gravis (MG) and Lambert--Eaton myasthenic syndrome (LEMS) through July 1998 was performed for the purpose of generating evidence-based practice parameters. There were 545 articles identified, of which 13 articles met at least three of the six criteria set previously by the American Association of Electrodiagnostic Medicine (AAEM). An additional 21 articles were identified from review articles or the references of these first 13 articles leading to a total of 34 articles. Results of studies utilizing repetitive nerve stimulation (RNS) showed that a 10% decrement in amplitude from the first to fourth or fifth intravolley waveform while stimulating at 2--5 HZ is valid for the diagnosis of MG. The degree of increment needed for the diagnosis of LEMS is at least 25% but most accurate when greater than 100%. Abnormal jitter or impulse blocking are the appropriate criteria for diagnosis of neuromuscular junction (NMJ) disorders when using single fiber electromyography (SFEMG). SFEMG is more sensitive than RNS for the diagnosis of disorders of neuromuscular transmission, but may be less specific and may not be available. Therefore, RNS remains the preferred initial test for MG and LEMS. PMID- 11494282 TI - Electrophysiologic correlates of anomalous course of the median-nerve thenar motor branch. PMID- 11494283 TI - What can AAEM members do about fraud? American Association of Electrodiagnostic Medicine. PMID- 11494284 TI - Derivated fetal haemoglobin as a marker for red cell age in the human fetus reflecting stimulated or impaired red blood cell production. AB - We have determined whether derivated fetal haemoglobin (dHbF, consisting of glycated and acetylated HbF) can be used as a cell age marker for fetal red blood cells (RBCs). Cord blood was obtained between 19 and 39 weeks of gestation from 28 alloimmunised anaemic fetuses (23 RhD+ and 5 Kell) and from 20 non-anaemic fetuses and newborns (controls). Density gradient centrifugation was applied to 36 samples (20 RhD+, 15 controls and 1 Kell) to obtain fractions of increasing cell age. Blood samples were used for measurements of mean cellular volume (MCV), mean cell haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), pyruvate kinase activity (PK) and derivated fetal haemoglobin (dHbF) by cation exchange HPLC. Reticulocytes were counted only in the whole blood samples. In all density gradient separated RBC fractions, the values for MCV, MCH and PK activity decreased and those of MCHC and dHbF increased with increasing density (equivalent to increasing cell age). The mean density was lower for RBCs of the anaemic RHD group (1.072+/-0.007 g/ml) than for the non-anaemic controls (1.077+/ 0.005 g/ml) (p<0.05) The RBC density of the Kell sensitised fetus did not differ from those of the controls. In the control group, the values of the cell age markers in whole blood changed significantly with the gestational age, showing an increase of mean age of the erythrocyte population. The best linear relationship was found for dHbF (y=6.28+0.17*weeks; r=0.84; p<0.001). In the anaemic RhD+ fetuses, the RBC age markers did not change with gestational age; the dHbF percentages were lower, and the MCV, MCH, PK values and the reticulocyte counts were higher than in the controls (0.05qter) and a partial monosomy of chromosome 16 (p13.3-->pter). PMID- 11494291 TI - Prenatal diagnosis in three cases of iniencephaly with unusual postmortem findings. AB - Iniencephaly is a rare and lethal congenital malformation of the neural tube characterized by occipital bone defect, cervical dysraphism, fixed retroflexion of the fetal head and severe lordosis of the cervicothoracic spine. The etiology is unknown. Prenatally diagnosed cases of iniencephaly are rare because careful and early ultrasonographic evaluation is necessary. We present three cases of iniencephaly prenatally diagnosed by sonography at 20-22 weeks' gestation in which therapeutic abortion was induced. The sonographic findings were compatible with the postmortem findings. The present cases of iniencephaly were found to carry unusual associated malformations such as two lobes in the right lung and chorangiosis of the placenta. Only hypoplastic lungs have been reported by previous authors. We also studied the 677C-->T mutation on the methylenetetrahydrofolate reductase gene in the parents in one of the present cases. The mother was found to be heterozygous for the 677CT polymorphism. PMID- 11494290 TI - Karyotypes found in the population declared at increased risk of Down syndrome following maternal serum screening. AB - Of the 65 328 pregnancies of South Australian mothers screened by the South Australian Maternal Serum Antenatal Screening (SAMSAS) Programme between 1 January 1991 and 31 December 1997, 3431 (5.25%) were declared at increased risk of fetal Down syndrome. Fetal or neonatal karyotype was determined in 2737/3431 (79.8%) of these pregnancies, including 16 with early fetal loss. Interrogation of the database of the South Australian Neonatal Screening Service showed 643 live-born infants whose phenotype was not subsequently questioned among the 694 pregnancies whose karyotype was not determined. Of the remaining 51/3431 pregnancies, 19 ended in early fetal loss without karyotyping and no newborn screening or other records could be found for 32 cases. The 129 instances of abnormal karyotype found were Down syndrome (84), trisomy 18 (four), trisomy 13 (three), triploidy (two), female sex chromosome aneuploidy (six) and male sex chromosome aneuploidy (five), inherited balanced rearrangements (19), mosaic or de novo balanced abnormalities (four) and unbalanced karyotypes (two). In the pregnancies declared at increased risk of fetal Down syndrome, only the karyotype for Down syndrome occurred with a frequency greater than that expected for the general, pregnant population. PMID- 11494292 TI - First trimester screening for Down syndrome and assisted reproduction: no basis for concern. AB - In pregnancies obtained after assisted reproduction the false-positive rate of second trimester Down syndrome (DS) screening is increased by 1.5-3-fold. This may cause an increase in the number of amniocenteses and the fetal loss rate. The present study for the first time examined whether assisted reproductive technologies affect the results of first trimester screening. The markers PAPP-A, free beta-hCG and the nuchal translucency (NT) thickness were examined at 12-14 weeks' gestation. Screening markers in 47 in vitro fertilisation (IVF), 63 ovulation induction (OI) and 3026 spontaneously conceived singleton pregnancies were compared. The MoM (multiples of the median) value in the IVF pregnancies was 1.02 (95% CI: 0.85-1.22) for PAPP-A, 1.14 (95% CI: 0.95-1.37) for beta-hCG and 0.97 (95% CI: 0.89-1.05) for NT; the MoM value in the OI pregnancies was 0.89 (95% CI: 0.76-1.05) for PAPP-A, 1.08 (95% CI: 0.93-1.25) for beta-hCG and 1.02 (95% CI: 0.95-1.11) for NT. The first trimester marker values in assisted reproductive pregnancies and spontaneously conceived pregnancies were not significantly different. Estimated false-positive rates for a risk cut-off of 1:400 varied from 4.7% in IVF pregnancies to 5.1% in OI pregnancies. Therefore the false-positive rate in Down syndrome screening should be independent of the method of conception. PMID- 11494294 TI - Maternal serum activin A and inhibin A in trisomy 18 pregnancies at 10-14 weeks. AB - In 45 cases of trisomy 18 and 493 control pregnancies at 10-14 weeks of gestation, maternal serum inhibin A, total activin A, free beta-hCG and PAPP-A were measured. In the trisomy 18 pregnancies the median values were 0.74 MoM for inhibin A, 1.23 MoM for activin A, 0.38 MoM for free beta-hCG and 0.16 MoM for PAPP-A. The degree of deviation from normal in the levels of inhibin and activin is small in comparison with free beta-hCG and PAPP-A and they are therefore unlikely to be of value in improving the sensitivity of 90% for a 1% false positive rate achieved by screening with fetal nuchal translucency and maternal serum free beta-hCG and PAPP-A. PMID- 11494293 TI - Microsatellite analysis provides efficient confirmation of fetal trophoblast isolation from maternal circulation. AB - Fetal trophoblasts can be found in maternal circulation from an early stage of pregnancy and thus provide a potential source of DNA for non-invasive prenatal diagnosis. We have developed a two-step method for trophoblast isolation between the 8th and 12th week of pregnancy. Blood was sampled from 14 women undergoing termination of pregnancy or spontaneous abortion. Immunomagnetic beads precoated with HLA class I and II, and with anti-cytokeratin-18 monoclonal antibodies, were used to remove CD8+ and other maternal cells, and to select for fetal trophoblasts, respectively. Microsatellite analysis was performed on DNA extracted from the isolated, maternal, paternal and placental cells after PCR amplification. Recovery of the trophoblasts was confirmed in 13/14 cases (93%) by the identification of an identical microsatellite pattern for fetal and placental cells. Further evidence was the presence of heterozygous alleles of both maternal and paternal origin. The correct prediction of gender in all five male fetuses was an additional confirmation of trophoblast recovery. We conclude that trophoblasts can be effectively isolated from maternal blood in the first trimester, and by using polymorphic microsatellite markers to confirm sample purity, this method has potential future application in prenatal diagnosis. PMID- 11494295 TI - Early ultrasound diagnosis of Neu-Laxova syndrome. AB - We report the mid-trimester prenatal diagnosis of Neu-Laxova syndrome (NLS) in two at risk families utilizing serial sonographic examinations. Ultrasound and pathologic findings from seven affected pregnancies, the largest case series of NLS to date, are presented. One fetus had anencephaly and incomplete rachischisis, an anomaly that has not been previously reported in association with NLS. Ultrasonographic detection of severe intrauterine growth restriction (IUGR), abnormally postured limbs, microcephaly, and edema allowed prenatal diagnosis of NLS in five of these at risk pregnancies during the mid-trimester. Growth curves derived from serial sonograms reveal abnormalities of all standard biometric measurements. The growth discrepancy was most pronounced in the measurements of the biparietal diameter, which were consistently less than two standard deviations below the mean across all gestational ages. This case series confirms that aberrant growth and anomalies may be detected sufficiently early in gestation to permit prenatal diagnosis of NLS. PMID- 11494296 TI - Restrictive dermopathy and fetal behaviour. AB - We report three siblings from consecutive pregnancies affected with restrictive dermopathy (RD). During the second pregnancy, fetal behavioural development and growth were studied extensively using ultrasound at 1-4 week intervals. Dramatic and sudden changes occurred in fetal body movements and growth but not until the end of the second trimester of pregnancy. Prominent at that time were prolonged periods of fetal quiescence and very low heart rate variability, together with abnormally executed body movements of short duration. Retarded femoral development and jerky abrupt fetal body movements (abnormal movement quality) were already present in the early second trimester of pregnancy. Facial anomalies emerged despite the presence of fetal mouth movements. The clinical features of RD were only partly explained by present knowledge of skin development and the fetal akinesia deformation sequence hypothesis. Quantitative assessment of fetal movements proved to be a poor early marker for antenatal diagnosis of this disorder. PMID- 11494297 TI - Prenatal evaluation of fetal renal function based on serum beta(2)-microglobulin assessment. AB - The relationship between fetal renal function (FRF) and fetal serum beta(2) microglobulin (B2MG) was investigated by comparing its value in 112 unaffected fetuses with that of 23 fetuses presenting with urinary tract malformations (UTM). Fetal serum level of B2MG was totally unrelated to gestational age; its value increased in cases of severe impairment of FRF but was similar to controls in all mild uropathies (p<0.05). Evaluating serum B2MG could be beneficial in fetuses with severe renal damage, but is of no use in unilateral UTM since only the global FRF is tested and not the function of each single kidney. PMID- 11494298 TI - Isolated mild fetal cerebral ventriculomegaly: a retrospective analysis of 26 cases. AB - We retrospectively studied 26 fetuses with isolated mild cerebral ventriculomegaly diagnosed between 1992 and 1998 and defined by a lateral ventricular atrial diameter of 10-15 mm without any other cerebral anomaly. Our objectives were to determine maternal risk factors, to evaluate complementary investigations, to assess developmental prognosis and to propose possible management. During pregnancy 10/26 patients had regressive ventriculomegalies, ten remained borderline at birth and six were confirmed postnatally. No maternal risk factors were identified. Prenatal investigations were carried out in 69% of cases but in only a few cases supplied any information. Postnatal examinations revealed one case of Down syndrome and one of porencephaly. Four children were lost to follow-up. In the 22 other cases, four had developmental delay. Early and unexplained mild ventriculomegaly appears to have a good prognosis. If ventriculomegaly is persistent, prenatal management should be carried out to investigate chromosomal abnormalities, viral infection, and fetal cerebral parenchymal damage. A long postnatal clinical follow-up is required. PMID- 11494299 TI - Three- versus two-dimensional ultrasound for nuchal translucency thickness measurements: comparison of feasibility and levels of agreement. AB - We have determined feasibility and levels of agreement for visualizing the nuchal translucency within a time limit with three-dimensional (3D) and two-dimensional (2D) transvaginal ultrasound. A total of 213 women of median gestational age 13 weeks (range 11-14 weeks) undergoing first-trimester nuchal translucency (NT) thickness measurements with 2D transabdominal ultrasound were included in the study. Additionally all women underwent 2D and 3D transvaginal NT examination within a time limit of 90 s. These two methods were compared with each other and with our standard method (2D transabdominal ultrasound) with respect to visualization of fetal nuchal fold and to the levels of agreement. The nuchal fold was visualized by 3D and 2D transvaginal ultrasound in 85.9% (95% CI: 80.5%, 90.3%) and 25.8% (95% CI: 20.1%, 32.2%), respectively (p<0.001). There was a statistically significant underestimation of the NT by 2D transabdominal as compared with 3D transvaginal ultrasound of 0.1 mm (p<0.001), and by 3D transvaginal as compared with 2D transvaginal ultrasound of even 0.1 mm (p<0.001). 3D transvaginal ultrasound of the nuchal fold has increased feasibility to 2D transvaginal ultrasound within a short examination time and with minimal, but significant, measurement differences. PMID- 11494303 TI - Current awareness in prenatal diagnosis. PMID- 11494300 TI - Prenatal diagnosis of respiratory chain deficiency by direct mutation screening. AB - Respiratory chain deficiency (RCD) is responsible for a clinically heterogeneous group of early-onset untreatable disorders. Enzymological prenatal diagnosis (PD) can only be offered to a fraction of families. Moreover, due to the two-fold genetic origin of the respiratory chain (nuclear and mitochondrial DNA) and owing to the large number of nuclear genes involved in the respiratory chain assembly, maintenance and functioning, the identification of the disease causing gene in a given family remains challenging. Here, we report on PD of RCD by direct screening of NDUFV1, SDH-Fp, SCO1 and SURF1 mutations in five unrelated families with complex I, II and IV deficiency, respectively. The identification of the disease-causing gene in a given family with RCD is a major issue to provide both adequate genetic counselling and early, reliable PD. PMID- 11494304 TI - Effects of Se, Cu and Se + vitamin E deficiency on the activities of CuZnSOD, GSH Px, CAT and LPO levels in chicken erythrocytes. AB - Antioxidant enzymes and vitamins provide a defence against the damage of cells by reactive oxygen species in living systems. The effect of Cu, Se and vitamin E deficiencies on the antioxidant enzyme activities and lipid peroxide levels of chicken erythrocytes were investigated during 6 weeks of a depletion diet. CuZnSOD activity and the plasma Cu level of the Cu-deficient group which was fed a diet containing 0.2 mg Cu x kg(-1) were reduced to 62 and 71% respectively. GSH Px activity of the Se-deficient group was decreased by 46% but by 21% in the Cu deficient group. CAT activity values of Se- and Cu-deficient groups were increased by 28 and 10% respectively. The maximum increase of LPO levels in erythrocyte membranes was observed as 32% for the Se+E-deficient group. The LPO level of the Cu-deficient group which had decreased CuZnSOD and GSH-Px activity, was also observed to be significantly increased when compared with the controls (p < 0.05). PMID- 11494305 TI - Effects of retinoids and thiazolidinediones on proliferation, insulin release, insulin mRNA, GLUT 2 transporter protein and mRNA of INS-1 cells. AB - Both 9-cis-retinoic acid (9cRA) and all-trans-retinoic acid (ATRA) are active metabolites of vitamin A (retinol). There exists an interaction between retinoid receptors and peroxisome proliferator-activated receptors (PPARgamma). To define their functions in an insulin secreting system the effects of ATRA, 9cRA and the PPARgamma agonist rosiglitazone on cell proliferation, insulin release and glucose transporter (GLUT) 2 of INS-1 cells were tested. Retinoic acid receptor (RAR-alpha and -gamma) and retinoid X receptor (RXR-alpha and -beta) proteins are present (immunoblots). Both 9cRA and ATRA inhibit INS-1 cell proliferation ([3H] thymidine assay) in a concentration dependent manner. Both 9cRA and ATRA increased insulin release, but only ATRA ralsed the GLUT 2 mRNA in a bell-shaped concentration response curve after 48 h. The insulinotropic effect of one compound is not significantly superimposed by the other indicating that the same binding sites are used by 9cRA and ATRA. The acute and chronic effects of the PPARgamma agonist rosiglitazone on insulin release were additionally determined since glitazones act as transcription factors together with RXR agonists. At high concentrations (100 microM) rosiglitazone inhibited glucose (8.3 mM) stimulated insulin secretion (acute experiment over 60 min). Insulin secretion, however, was increased during a 24 h treatment at a concentration of 10 microM and again inhibited at 100 microM. Changes in preproinsulin mRNA expression were not observed. Rosiglitazone (100 microM) increased GLUT 2 mRNA paralleled by an increase of GLUT 2 protein, but only after 24 h of treatment. This data indicate that RAR and RXR mediate insulin release. The changes in GLUT 2 have no direct impact on insulin release; the inhibition seen at high concentrations of either compound is possibly the result of the observed inhibition of cell proliferation. Effects of rosiglitazone on preproinsulin mRNA and GLUT 2 (mRNA and protein) do not play a role in modulating insulin secretion. With the presence of an RXR receptor agonist the effect of rosiglitazone on insulin release becomes stimulatory. Thus the effects of RAR-, RXR agonists and rosiglitazone depend on their concentrations, the duration of their presence and are due to specific interactions. PMID- 11494306 TI - Pancreatic fate of D-[3H] mannoheptulose. AB - D-Mannoheptulose was recently postulated to be transported into cells by GLUT2. The validity of such an hypothesis was assessed by comparing the uptake of tritiated D-mannoheptulose by pancreatic islets versus pieces of pancreas and, in the latter case, by comparing results obtained in control rats versus animals injected with streptozotocin (STZ). The uptake of D-[3H] mannoheptulose by islets represents a time-related and temperature-sensitive process, inhibited by cytochalasin B and enhanced by D-glucose. The uptake of the tritiated heptose was much lower in pieces of pancreatic tissue and inhibited by D-glucose, at least in the STZ rats. Whether in pieces of pancreas exposed in vitro to D-[3H] mannoheptulose or after intravenous injection of the tritiated heptose, the radioactive content of the pancreatic tissue was lower in STZ rats than in control animals. This contrasted with an unaltered radioactive content of liver and muscle in the STZ rats, at least when treated with insulin. Suitably radiolabelled D-mannoheptulose or an analogue of the heptose could thus conceivably be used for quantification of the endocrine pancreatic mass. PMID- 11494307 TI - Antimicrotubular and cytotoxic activity of geiparvarin analogues, alone and in combination with paclitaxel. AB - Geiparvarin is an antiproliferative compound isolated from the leaves of Geijera parviflora, and may represent a new drug which targets tubulin. To better explore the potential use of this agent, we investigated the antimicrotubular and cytotoxic effects of new synthetic aromatic derivatives of geiparvarin. These drugs inhibited polymerization of microtubular protein, particularly when the assembly was induced by paclitaxel. The microtubular network organization of fibroblasts was altered more effectively by some drugs. Normal microtubule architecture completely disappeared when the cells were treated simultaneously with drugs and paclitaxel: microtubules depolymerized or were reorganized into bundles, in a similar but more disarrayed fashion than that observed after treatment with paclitaxel alone. Cytotoxicity studies showed a dose-dependent effect, whereas combined administration of drugs and paclitaxel increased cytotoxicity, more effectively in paclitaxel versus derivatives administration alone. PMID- 11494308 TI - Is epsilon4 allele of apolipoprotein E associated with more severe end-organ damage in essential hypertension? AB - The aim of the present study comparing patients with mild to moderate hypertension with controls, was to explore a possible association between hypertension-related target organ damage and evaluation found in the gene encoding apolipoprotein E (apo E) genotype. Detailed medical history was recorded and physical examination was performed for all patients in the study (88 hypertensives, 63 normotensive controls). PCR (Polymerase Chain Reaction), RFLP (Restriction Fragment Length Polymorphism), and agarose gel electrophoresis techniques were used to determine the apo E genotypes. The frequencies of apo epsilon2, apo epsilon3, and apo epsilon4 alleles were 3.97, 88.06, and 9.95%, respectively in the hypertensive group. The frequencies of apo epsilon2, apo epsilon3, and apo epsilon4 alleles were 5.5, 92.0, and 2.38%, respectively in the control group. There were about twice as many individuals in the heterozygote hypertensive group who had apo E3/4 as compared to the control group (7.30 vs. 2.38%) (p = 0.07). The hypertensive patients who were carriers of the apo epsilon4 had significantly higher organ damage (left ventricular hypertrophy (p < 0.001). dilated left atrium (p < 0.05), retinopathy (p < 0.05)) as compared to those who were not carriers of apo epsilon4. These results showed a trend for the epsilon4 allele to be associated with a higher prevalence of target organ damage in patients with mild to moderate hypertension. PMID- 11494309 TI - Effects of phosphoprotein phosphatase inhibitors (phenylarsine oxide and cantharidin) on Tetrahymena. AB - The effects of phenylarsine oxide (PAO) (phosphotyrosine phosphatase inhibitor) and cantharidin (serine/threonine phosphatase [PP2A] inhibitor) treatments were analysed on the synthesis of phospholipids and glycolipids, and on the cytoskeletal elements (F-actin and tubulin containing structures) of Tetrahymena pyriformis. Both phosphatase inhibitors reduced the amount of incorporated 32P of the whole phospholipid content, but the ratio of phosphatidylserine (PS) and phosphatidylcholine (PC) to the total phospholipid content increased. Both treatments influenced the phosphatidylinositol (PI) system. These inhibitors also influenced the incorporation of palmitic acid into the phospholipids: in general PAO decreased, whereas cantharidin increased the amount of incorporated palmitic acid; 1 microM cantharidin significantly increased the labelling of PE and PA. The incorporation of mannose and glucosamine was influenced differently by PAO and cantharidin treatments: the latter elevated, while PAO decreased the labelling of glycolipids with these sugars. The effects of these treatments were visible also in the case of confocal scanning laser microscopic (CSLM) images: after treatments with both inhibitors, the F-actin containing cortical elements were destroyed, but the tubulin containing ones (longitudinal and transversal microtubules, oral apparatus and deep fibres) did not display significant alterations. The different effects of phosphatase inhibitors were visible also on the scanning electron microscopic (sEM) images: cantharidin treatments (1 microM) decreased the amount of dissolved membrane lipids after chemical dehydration of the cells with 2, 2-dimethoxy propane (DMP), but in the case of treatments with 10 microM, the surface pattern of cells was similar to the controls. On the other hand, after PAO treatments the surface pattern of Tetrahymena showed significant alterations. Both phosphatase inhibitors inhibited the phagocytotic activity of the cells. On the basis of present experiments we suppose that these treatments are able to influence signalling systems (e.g. PI) of Tetrahymena, and also the structure of the cytoskeleton and the functions (e.g. phagocytosis) which are connected with skeletal elements. PMID- 11494310 TI - Tissue malondialdehyde and adenosine triphosphatase level after experimental liver ischaemia-reperfusion damage. AB - Functional irregularities due to damage after ischaemia-reperfusion vary depending upon the organs affected. High energy phosphates such as ATP and ADP are destroyed after ischaemia-reperfusion damage. Subsequently, protons and inorganic phosphates accumulate within the cells and the proton pumps such as adenosine triphosphatase (ATPase), which maintain intracellular ion balance are damaged. In the present study, malondialdehyde (MDA), a product of lipid peroxidation, was measured as an indicator of tissue damage. Additionally, we measured sodium-potassium-ATPase levels and determined the interactions between MDA and Na+-K+ ATPase levels. A total of 31 female guinea pigs were divided into four groups: sham operated guinea pigs (group 1), ischaemia-reperfusion (group 2), ischaemia-reperfusion + superoxide dismutase (SOD) (group 3), ischaemia reperfusion + allopurinol (group 4). Following reperfusion, the livers of guinea pigs in each group were removed for histopathological examination and the levels of MDA and Na+-K+ ATPase were determined in homogenized tissue samples. There was a statistically significant (p < 0.05) reduction in tissue MDA levels in group 2 when compared with group 1. The level of tissue MDA in groups 3 and 4 was significantly lower than tissue MDA levels of group 2. However, there was a statistically significant (p < 0.05) reduction in tissue Na+-K+ ATPase levels of group 2 when compared with group 1. Similarly, the level of tissue Na+-K+ ATPase in groups 3 and 4 was significantly higher than the tissue Na+-K+ ATPase levels of group 2. The results of the histopathologic examination also revealed the beneficial effects of the use of SOD and allopurinol in preventing liver damage in cases of ischaemia-reperfusion. Although the levels of MDA and Na+-K+ ATP ase in group 2 were not equal to the level in group 1, antioxidant therapy significantly improved the tendency to reverse the effects of ischaemia reperfusion and to protect the liver from damage due to ischaemia-reperfusion. PMID- 11494311 TI - Suppression of EGF-induced cell proliferation by the blockade of Ca2+ mobilization and capacitative Ca2+ entry in mouse mammary epithelial cells. AB - The role of intracellular Ca2+ stores and capacitative Ca2+ entry on EGF-induced cell proliferation was investigated in mouse mammary epithelial cells. We have previously demonstrated that EGF enhances Ca2+ mobilization (release of Ca2+ from intracellular Ca2+ stores) and capacitative Ca2+ entry correlated with cell proliferation in mouse mammary epithelial cells. To confirm their role on EGF induced cell cycle progression, we studied the effects of 2,5-di-tert butylhydroquinone (DBHQ), a reversible inhibitor of the Ca2+ pump of intracellular Ca2+ stores, and SK&F 96365, a blocker of capacitative Ca2+ entry, on mitotic activity induced by EGF. Mitotic activity was examined using an antibody to PCNA for immunocytochemistry. SK&F 96365 inhibited capacitative Ca2+ entry in a dose-dependent manner (I50: 1-5 microM). SK&F 96365 also inhibited EGF induced cell proliferation in the same range of concentration (I50: 1-5 microM). DBHQ suppressed [Ca2+]i response to UTP and thus depleted completely Ca2+ stores at 5 microM. DBHQ also inhibited EGF-induced cell proliferation at an I50 value of approximately 10 microM. The removal of these inhibitors from the culture medium increased the reduced mitotic activity reversibly. Using a fluorescent assay of DNA binding of ethidium bromide, no dead cells were detected in any of the cultures. These results indicate that the inhibitory effects of SK&F 96365 and DBHQ on cell proliferation were due to the inhibition of capacitative Ca2+ entry and Ca2+ mobilization suggesting the importance of capacitative Ca2+ entry and Ca2+ mobilization in the control of EGF-induced cell cycle progression in mouse mammary epithelial cells. PMID- 11494312 TI - Studies on p53 and Bax protein expression in Cockayne syndrome cells after UV irradiation and interferon-beta treatment. AB - Human interferon (HuIFN) has a protective effect against ultraviolet (UV)-induced killing of Cockayne syndrome (CS) and xeroderma pigmentosum (XP) cells. Irradiation with ultraviolet (UV) resulted in nuclear accumulation of p53 in normal human fibroblast cells, and this accumulation was suppressed by treatment with HuIFN-beta. On the other hand, a large amount of p53 was found in both nuclear and cytoplasmic fractions of one SV40-transformed XP and two SV40 transformed CS cell strains irrespective of UV irradiation. Treatment with HuIFN beta reduced the level of pro-apoptotic Bax protein without suppression of nuclear accumulation of p53 in the CS cells but not in the XP cells. These findings suggest that there are different mechanisms of UV-refractoriness caused by HuIFN-beta in UV-sensitive CS and XP cells. PMID- 11494313 TI - Hot news: temperature-sensitive humans explain hereditary disease. AB - The skin-cancer-prone hereditary disease xeroderma pigmentosum is typically characterized by defective nucleotide excision repair (NER) of DNA. However, since all subunits of the core basal transcription factor TFIIH are required for both RNA polymerase II basal transcription and NER, some mutations affecting genes that encode TFIIH subunits can result in clinical phenotypes associated with defective basal transcription. Among these is a syndrome called trichothiodystrophy (TTD) in which the prominent features are brittle hair and nails, and dry scaly skin. A recent study provides dramatic support for the so called transcription hypothesis of TTD.(1) Specifically, several patients have been shown to carry a mutation in the XPD gene, which encodes a thermolabile form of XPD protein, resulting in loss of hair during febrile episodes. PMID- 11494314 TI - The role of chemoattraction in cancer metastases. AB - It has long been unclear as to why particular cancers preferentially metastasize to certain sites. The possibilities usually discussed involve differential survival and proliferation at these sites, or selective trapping with or without preferential homing. A recent report by Muller et al.(1) provides evidence for preferential homing of breast cancer to metastatic sites. The findings indicate that the chemokine receptors CXCR4 and CCR7 are found on breast cancer cells and their ligands are highly expressed at sites associated with breast cancer metastases. This results in chemotaxis, or directed migration of tumor cells from their primary site via the circulation to the preferential sites of metastases. The evidence for this model and its significance are reviewed here. PMID- 11494315 TI - Steroid-triggered death by autophagy. AB - Programmed cell death is a critical part of normal development, removing obsolete tissues or cells and sculpting body parts to assume their appropriate form and function. Most programmed cell death occurs by apoptosis of individual cells or autophagy of groups of cells. Although these pathways have distinct morphological characteristics, they also have a number of features in common, suggesting some overlap in their regulation. A recent paper by Lee and Baehrecke provides further support for this proposal.(1) These authors present, for the first time, a genetic analysis of autophagy, using the steroid-triggered metamorphosis of Drosophila as a model system. They demonstrate a remarkable degree of overlap between the control of apoptosis and autophagy as well as a key role for the steroid-inducible gene E93 in directing the autophagic death response. This paper also shows that E93 can direct cell death independently from the known death inducer genes, defining a novel death pathway in Drosophila. PMID- 11494316 TI - The CtBP family: enigmatic and enzymatic transcriptional co-repressors. AB - Transcription factors that associate with DNA sequences in promoters and enhancers often recruit co-regulators that modulate their activity. Many of these co-regulators have intrinsic enzymatic activity and influence gene expression by modifying chromatin and altering its structure. Recently, a new family of co repressors, the C-terminal binding proteins, has been described. These proteins recognize Pro-X-Asp-Leu-Ser (PXDLS) motifs in DNA-binding proteins and function as transcriptional co-repressors in Drosophila, Xenopus and mammals. The precise mechanisms by which they influence transcription are still under investigation. CtBP proteins dimerize and can contact histone deacetylases; hence they may operate by linking deacetylases to DNA-bound factors. But it appears that CtBP proteins also have intrinsic enzymatic activity. They have significant homology to D-isomer-specific 2-hydroxy acid dehydrogenases, and remarkably one family member, rat CtBP, has been shown to have a second role, functioning as an acyl transferase in Golgi maintenance. These observations raise the possibility that CtBP proteins might regulate gene expression directly by means of their enzymatic activities, in addition to serving as simple bridging proteins. Supplementary material for this article can be found on the BioEssays homepage at http://www.interscience.wiley.com/jpages/0265-9247/suppmat/v23_8.684. PMID- 11494317 TI - Drosophila peripodial cells, more than meets the eye? AB - Drosophila imaginal discs (appendage primordia) have proved invaluable for deciphering cellular and molecular mechanisms of animal development. By combining the accessibility of the discs with the genetic tractability of the fruit fly, researchers have discovered key mechanisms of growth control, pattern formation and long-range signaling. One of the principal experimental attractions of discs is their anatomical simplicity - they have long been considered to be cellular monolayers. During larval stages, however, the growing discs are 2-sided sacs composed of a columnar epithelium on one side and a squamous 'peripodial' epithelium on the other. Recent studies suggest important roles for peripodial epithelia in processes previously assumed to be confined to columnar cell monolayers. PMID- 11494318 TI - The development of the Drosophila genital disc. AB - The imaginal discs of Drosophila melanogaster, which form the adult epidermal structures, are a good experimental model for studying morphogenesis. The genital disc forms the terminalia, which are the most sexually dimorphic structures of the fly. Both sexes of Drosophila have a single genital disc formed by three primordia. The female genital primordium is derived from 8(th) abdominal segment and is located anteriorly, the anal primordium (10 and 11(th) abdominal segments) is located posteriorly, and the male genital primordium from the 9(th) abdominal segment lies between them. In both sexes, only two of these three primordia develop to form the adult terminalia. The anal primordium develops in both sexes but, depending on the genetic sex, will form either male or female analia. However, only one of the genital primordia develops in each sex, forming either the male or the female genitalia. This depends on the genetic sex of the fly. Therefore, the genital disc is a very good experimental model of how the sex determination and homeotic genes - which determine cell identity - interact to direct the development of a population of cells into male or female terminalia. It has been proposed that the sexually dimorphic development of the genital disc is the result of an integrated genetic input, made up by the sex-determination gene doublesex and the homeotic gene Abdominal-B. This input acts by modulating the response to Hedgehog, Wingless, and Decapentaplegic morphogenetic signals. PMID- 11494319 TI - Intrinsic DNA bends: an organizer of local chromatin structure for transcription. AB - DNA with a curved trajectory of its helix axis is called bent DNA, or curved DNA. Interestingly, biologically important DNA regions often contain this structure, irrespective of the origin of DNA. In the last decade, considerable progress has been made in clarifying one role of bent DNA in prokaryotic transcription and its mechanism of action. However, the role of bent DNA in eukaryotic transcription remains unclear. Our recent study raises the possibility that bent DNA is implicated in the "functional packaging" of transcriptional regulatory regions into chromatin. In this article, I review recent progress in bent DNA research in eukaryotic transcription, and summarize the history of bent DNA research and several subjects relevant to this theme. Finally, I propose a hypothesis that bent DNA structures that mimic a negative supercoil, or have a right-handed superhelical writhe, organize local chromatin infrastructure to help the very first interaction between cis-acting DNA elements and activators that trigger transcription. PMID- 11494320 TI - Hydra and Niccolo Paganini (1782-1840)--two peas in a pod? The molecular basis of extracellular matrix structure in the invertebrate, Hydra. AB - The body wall of Hydra is organized as an epithelial bilayer with an intervening extracellular matrix (ECM). Molecular and biochemical analyses of Hydra ECM have established that it contains components similar to those seen in more complicated vertebrates such as human. In terms of biophysical parameters, Hydra ECM is highly flexible; a property that facilitates continuous movements along the organism's longitudinal and radial axis. A more rigid ECM, as in vertebrates, would not be compatible with this degree of movement. The flexible nature of Hydra ECM can now be explained in part by the unique structure of the organism's collagens. Interestingly, some aspects of the structural features of Hydra collagens mimic what is seen in Ehlers-Danlos syndrome, an inherited condition in humans that results in an abnormally flexible ECM that can be debilitating in extreme cases. This review will focus on structure-function relationships of the ECM of Hydra. PMID- 11494321 TI - Cryo-electron microscopy as an investigative tool: the ribosome as an example. AB - Cryo-electron microscopy allows the visualization of macromolecules in their native state. Combined with techniques of three-dimensional reconstruction, cryo EM images of single molecules can be used to study macromolecular interactions. The ribosome, a large RNA-protein complex with multiple binding interactions, is an excellent test case illustrating the power of these new techniques. Conformational changes during the binding of tRNA and protein factors to the ribosome can now be studied without the interference of crystal packing. Now that the first X-ray structures of ribosomal subunits have become available, conformational changes observed by cryo-EM in different functional states can be traced back to internal rearrangements of the underlying structural framework. Electron microscopy, X-ray crystallography, and modeling should be used together in the endeavor to understand the functioning of the translational machinery. PMID- 11494322 TI - Presenilin mutations and calcium signaling defects in the nervous and immune systems. AB - Presenilin-1 (PS1) is thought to regulate cell differentiation and survival by modulating the Notch signaling pathway. Mutations in PS1 have been shown to cause early-onset inherited forms of Alzheimer's disease (AD) by a gain-of-function mechanism that alters proteolytic processing of the amyloid precursor protein (APP) resulting in increased production of neurotoxic forms of amyloid beta peptide. The present article considers a second pathogenic mode of action of PS1 mutations, a defect in cellular calcium signaling characterized by overfilling of endoplasmic reticulum (ER) calcium stores and altered capacitive calcium entry; this abnormality may impair synaptic plasticity and sensitize neurons to apoptosis and excitotoxicity. The calcium signaling defect has also been documented in lymphocytes, suggesting a contribution of immune dysfunction to the pathogenesis of AD. A better understanding of the calcium signaling defect resulting from PS1 mutations may lead to the development of novel preventative and therapeutic strategies for disorders of the nervous and immune systems. PMID- 11494323 TI - Securing genome stability by orchestrating DNA repair: removal of radiation induced clustered lesions in DNA. AB - In addition to double- and single-strand DNA breaks and isolated base modifications, ionizing radiation induces clustered DNA damage, which contains two or more lesions closely spaced within about two helical turns on opposite DNA strands. Post-irradiation repair of single-base lesions is routinely performed by base excision repair and a DNA strand break is involved as an intermediate. Simultaneous processing of lesions on opposite DNA strands may generate double strand DNA breaks and enhance nonhomologous end joining, which frequently results in the formation of deletions. Recent studies support the possibility that the mechanism of base excision repair contributes to genome stability by diminishing the formation of double-strand DNA breaks during processing of clustered lesions. PMID- 11494324 TI - Spider flagelliform silk: lessons in protein design, gene structure, and molecular evolution. AB - Spiders spin multiple types of silks that are renowned for their superb mechanical properties. Flagelliform silk, used in the capture spiral of an orb web, is one of the few silks characterized by both cDNA and genomic DNA data. This fibroin is composed of repeating ensembles of three types of amino acid sequence motifs. The predominant subrepeat, GPGGX, likely forms a beta-turn, and tandem arrays of these turns are thought to create beta-spirals. These spring like helices may be critical for the exceptional ability of capture silk to stretch and recoil. Each ensemble of motifs was found to correspond to a different exon within the flagelliform gene. The pattern of sequence similarity among exons indicates intragenic concerted evolution. Surprisingly, the introns between the iterated exons are also homogenized with each other. This unusual molecular architecture in the flagelliform silk gene has implications for the evolution and maintenance of spider silk proteins. PMID- 11494325 TI - The control of cultural evolution has been tried, what's next? PMID- 11494327 TI - In memoriam: Gary J. Miller,MD, PhD. PMID- 11494328 TI - Apoptotic activity of doxazosin on prostate stroma in vitro is mediated through an autocrine expression of TGF-beta1. AB - BACKGROUND: Doxazosin, an alpha-adrenergic antagonist, has been shown to induce apoptosis in prostatic stromal cells. The mechanism of this apoptotic action by Doxazosin remains undefined. The present study was carried out to demonstrate that the effect of Doxazosin on apoptosis of prostate stromal cells is mediated through an autocrine action of TGF-beta1. METHODS: Primary cultures of human prostate cells were treated with varying concentrations of Doxazosin (0, 0.1, 1, 10, and 100 microM) for a period up to 3 days. At the end of the 3-day culture, cell numbers were counted. Apoptosis was assessed by a colorimetric terminal deoxyribonucleotide transferase labeling technique. TGF-beta1 was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared to control cultures, cell numbers were significantly decreased as much as 68.4% in cultures treated with 10 microM of Doxazosin after 3 days incubation, while apoptosis increased by 64.7% in cultures treated with the same concentration of Doxazosin after 24 h. This decrease in cell number was reversed when antibody to TGF-beta1 was added to these cultures. Addition of TGF-beta1 (0, 1.0, and 10 ng/mL) to the cultures also decreased the cell numbers. Quantitation of TGF-beta1 in lysates of cells by ELISA revealed that the cells treated with Doxazosin (10 microM) produced as much as 62.5% more TGF-beta1 than in that of untreated cells. CONCLUSIONS: These results demonstrate that the apoptotic effect of Doxazosin on human prostatic stromal cells is mediated through an autocrine production of TGF-beta1. PMID- 11494329 TI - Impact of preoperative serum PSA level from 0 to 10 ng/ml on pathological findings and disease-free survival after radical prostatectomy. AB - BACKGROUND: To determine the impact of various preoperative serum prostate specific antigen (PSA) levels in the range from 0.1 to 10 ng/ml on pathological stage and disease-free survival after radical prostatectomy. METHODS: We selected a cohort of 585 patients who underwent radical prostatectomy between 1991-1996 for clinically localized prostate cancer and presented with preoperative serum PSA levels from 0.1 to 10 ng/ml. RESULTS: Pathological organ-confined disease was present in 57.6% of patients. The rate of organ-confined disease decreased from an average of 85% for patients with a PSA value < 2 ng/ml, to 46.8% for patients with a PSA value > 7 ng/ml. We found statistically significant correlations between preoperative serum PSA level and overall pathological stage (P = 0.001), pathologically organ-confined disease (P = 0.001), margin positive rates (P = 0.001), extra prostatic extension (P = 0.001), and seminal vesicle invasion (P = 0.001). The overall disease-free survival rate was 87%, with a median follow up of 42.4 months. Disease free survival was significantly better for patients with PSA up to 4 ng/ml (P = 0.005). CONCLUSIONS: Our data suggests that PSA detection programs should strive to detect prostate cancer in men before the PSA level rises above 7 ng/ml. In addition, since patients with a PSA level < 4 ng/ml had better disease-free survival rates than those with a PSA level between 4.1-10 ng/ml, eliminating an arbitrary cutoff of 4 ng/ml, may lead to improved disease free survival. PMID- 11494330 TI - Karyometry of secretory cell nuclei in high-grade PIN lesions. AB - BACKGROUND: The goal of this study was a karyometric characterization of secretory cell nuclei in high-grade prostatic intraepithelial neoplasia (PIN) lesions. Specifically, the hypothesis is tested that distinctly different subgroups of nuclei exist in these lesions. METHODS: High-resolution images of 1,713 nuclei from high-grade PIN lesions were recorded. Karyometric features were computed. Discriminant function scores against normal reference nuclei, and nuclear abnormality values were derived. Data sets were processed by a nonsupervised learning algorithm to establish the presence of subgroups of nuclei with statistically different nuclear chromatin distributions. RESULTS: Three sets of nuclei were formed, facing an intact basal cell layer, a near vanishing basal cell layer, and a gap in the basal cell layer. For each set, a nonsupervised learning algorithm formed three statistically different subgroups of approximately equal sizes. Each subgroup is found in every one of the three sampling locations. The total optical density distribution of nuclei in two subgroups suggests an aneuploid distribution, the third subgroup has a near diploid distribution. CONCLUSIONS: Secretory cell nuclei in high-grade PIN lesions are a heterogeneous population, forming statistically different subgroups. Studies aimed at characterizing the progression of such lesions should consider the inhomogeneous nature of these nuclei. PMID- 11494331 TI - Primary culture of microvascular endothelial cells from human benign prostatic hyperplasia. AB - BACKGROUND: Prostate growth seems to be influenced by paracrine factors like IL-6 originating from the microvascular endothelium. Therefore, our efforts were focused on the primary culture and behavior of microvascular endothelial cells (HPEC) derived from tissue of human benign prostatic hyperplasia (BPH). Until now, the isolation and culture of HPEC from BPH have not been reported. METHODS: BPH tissue was cut into small cubes and gently squeezed after incubation with dispase. HPEC were cultured from the resulting cell suspension after a stepwise selection by use of superparamagnetic beads coated with antibodies against endothelial specific antigens. HPEC were characterized by flow cytometry and immunohistochemistry. gamma-Glutamyl transpeptidase activity (specific for microvascular endothelium) was measured after dissolution of the HPEC with Triton X-100. After the incubation of HPEC either with ATP, VEGF, or TNF-alpha, the release of IL-6 was measured by enzyme linked immunosorbent assay (ELISA). RESULTS: HPEC showed a typical endothelial morphology. They were positive for von Willebrand factor, CD31, CD62E (after stimulation with TNF-alpha), alpha-actin and were negative for fibroblastic antigens and PSA. Proliferation was stimulated by vascular endothelial growth factor (VEGF). gamma-Glutamyl transpeptidase activity in HPEC was 6.3 microIU/microg protein, whereas in human umbilical vein endothelial cells (HUVEC) no gamma-glutamyl transpeptidase activity was detectable. The IL-6 secretion of HPEC was stimulated by VEGF and TNF-alpha, but not by ATP and bradykinin. CONCLUSIONS: For the first time, the primary culture of microvascular endothelial cells from BPH tissue was successfully performed. Our results suggest that HPEC may be actively involved in prostate growth, due to the secretion of regulatory factors such as IL-6. PMID- 11494332 TI - Expression profiles of voltage-gated Na(+) channel alpha-subunit genes in rat and human prostate cancer cell lines. AB - BACKGROUND: Voltage-gated Na(+) channel (VGSC) activity has been implicated in prostate cancer (PC) metastasis. Although VGSCs can occur as multiple-subunit assemblies, the alpha-subunits (VGSCalphas) alone can encode functional channels. The VGSCalpha gene(s) responsible for the functional VGSCalpha expression in strongly metastatic PC cell lines is not known. METHODS: Two reverse transcription-PCR (RT-PCR) methods, degenerate primer screening and a novel semi quantitative PCR (SQT-PCR) technique, were used. These methods enabled a detailed qualitative and quantitative investigation of VGSCalpha mRNA expression in rat (MAT-LyLu/AT-2) and human (PC-3/LNCaP) PC cells of markedly different metastatic potential. RESULTS: Expression of eight different VGSCalpha genes (SCN1A-4A, SCN7A-9A, and SCN11A) was determined in the PC cell lines. Most were expressed as multiple splice variants. SQT-PCR results were consistent with a basal level of VGSCalpha mRNA expression occurring in weakly metastatic (AT-2/LNCaP) cells, and this being greatly elevated in cells of stronger metastatic potential (MAT LyLu/PC-3), primarily due to the elevated expression of the SCN9A gene (also termed PN1/hNe-Na). CONCLUSIONS: (1) Several VGSCalpha genes and their splice variants are expressed similarly in both rat and human PC cell lines. (2) Expression levels are much higher in the strongly metastatic (MAT-LyLu/PC-3) cells. (3) Levels of SCN9A mRNA specifically are predominant in MAT-LyLu and PC-3 cells; thus, SCN9A is highly likely to be the main source of the functional VGSC detected. PMID- 11494333 TI - Histamine H(1) receptor activation inhibits the proliferation of human prostatic adenocarcinoma DU-145 cells. AB - BACKGROUND: Histamine stimulates cell proliferation in some tumor cell lines through the activation of H(1) receptors coupled to phosphoinositide hydrolysis. We therefore set out to study the presence of H(1) receptors in the prostate cancer cell line DU-145 and the effect of their stimulation on cell growth. METHODS: The presence of histamine receptors was studied by radioligand binding. Phosphoinositide hydrolysis was assessed by measuring [(1)H]-inositol phosphate ([(1)H]-IPs) accumulation and changes in the intracellular concentration of free Ca(2+) ([Ca(2+)](i)). Proliferation was assessed by cell counting and by [(1)H] thymidine incorporation. RESULTS: DU-145 cells express H(1) receptors (110+/-14 fmol/mg of protein) whose stimulation results in [(1)H]-IPs accumulation (602+/ 23% of basal, EC(50) 2.2+/-0.4 microM) and calcium mobilization (resting level 96+/-5 nM, Delta[Ca(2+)](i) 517+/-32 nM, EC(50) 6.2+/-0.1 microM). Incubation with histamine (100 microM, 24 hr) resulted in a decrease in both cell number and [(1)H]-thymidine incorporation, blocked by the H(1) antagonist mepyramine (1 microM). CONCLUSIONS: Histamine inhibits the proliferation of DU-145 cells through the activation of H(1) receptors coupled to phosphoinositide hydrolysis. PMID- 11494334 TI - Regulation of prostate-specific antigen (PSA) gene expression and release in LNCaP prostate cancer by antagonists of growth hormone-releasing hormone and vasoactive intestinal peptide. AB - BACKGROUND: Prostate-specific antigen (PSA) is the best tumor marker for diagnosis and prognosis of prostatic carcinoma. The secretion of PSA from LNCaP human prostate cancer cells is influenced by acute stimuli such as vasoactive intestinal peptide (VIP), growth hormone-releasing hormone (GHRH), and chronic stimuli like androgens. METHODS: To study the regulation of basal and VIP/GHRH or androgen-stimulated secretion from LNCaP cells, we used a superfusion system, which allowed us to simultaneously measure PSA gene expression, PSA secretion, and cAMP release from the same cancer cells. LNCaP cancer cells were also implanted orthotopically into nude mice. RESULTS: VIP (30 pM-3 nM), GHRH (3 nM 300 nM), and dihydrotestosterone (100 nM) induced a significant increase in PSA gene expression, PSA secretion, and cAMP release. The dose and time-dependent effects of peptides were manifested only in the presence of androgens. At the end of continuous stimulation of cells with 1 nM VIP for 2 hr, large amounts of stored immunoreactive PSA still remained in the cells. Adenylate cyclase activator, forskolin (FSK), significantly increased PSA secretion and gene expression, and potassium, which causes nonspecific depolarization of membranes, augmented gene expression, and secretion of PSA, but did not influence cAMP release. This suggests that PSA secretion is regulated by cAMP-dependent as well as cAMP-independent pathways. In superfusion system, stimulatory effects of VIP and GHRH on PSA secretion were inhibited by VIP antagonist JV-1-53, and less by GHRH antagonist JV-1-38. In cell cultures, JV-1-38 had a stronger inhibitory effect on proliferation, indicating an involvement of the recently discovered tumoral GHRH receptors in this process. In nude mice, with orthotopically implanted LNCaP cancer cells, GHRH antagonist JV-1-38 alone or androgen ablation by castration had no effect on tumor growth and PSA levels. However, castration combined with treatment with GHRH antagonist, significantly decreased tumor growth and PSA secretion. CONCLUSIONS: Our findings suggest that the secretion of PSA is regulated rather than constitutive, contrary to previous reports. In addition, the effect of GHRH and VIP antagonists on PSA secretion from prostate cancer cells is not correlated with their antiproliferative action. PMID- 11494335 TI - Polymorphic repeats in the androgen receptor gene in high-risk sibships. AB - BACKGROUND: Genetic susceptibility may explain some familial clusters of prostate cancer. The polymorphic androgen receptor (AR) gene, which mediates androgen activity in the prostate, is a candidate gene that may influence predisposition to the disease. METHODS: We analyzed the polymorphic (CAG)n and (GGN)n repeats within the AR gene in men from 51 high-risk prostate cancer sibships, which included at least one affected and one unaffected man (n = 210). We compared repeat lengths of men with prostate cancer (n = 140) to their brothers (n = 70) without disease, stratified by median age at diagnosis of affected men within each sibship. Conditional logistic regression was used to compute odds ratios (OR) and 95% confidence intervals to evaluate associations between prostate cancer and repeat length. RESULTS: The OR for prostate cancer associated with short (CAG)n repeats (< 22) compared to longer repeats (> or =22) was 1.13 (95% CI 0.5-2.4) overall, but was higher in sibships with a median age of <66 years at diagnosis (OR = 1.72, 95% CI 0.5-6.0). The (GGN)n array also was not associated with prostate cancer in general. However, in older men (> or = 66 years), there was a modest elevation in risk (OR = 1.56, 95% CI 0.6-4.1) among those with short repeats (GGN of < or =16). Men with both a short (CAG)n (< 22) and a short (GGN)n (< or =16) array were not at higher risk (OR = 1.06) compared to men with two long repeats [(CAG)n > or =22 and (GGN)n >16)]. CONCLUSIONS: These results suggest that the (CAG)n and (GGN)n repeats in the AR gene do not play a major role in familial prostate cancer. PMID- 11494336 TI - Expression of the matrix metalloproteinase promatrilysin in coculture of prostate carcinoma cell lines. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are involved in tumor progression. Matrilysin (MMP-7) has been shown to be upregulated in prostatic carcinomas and can increase the invasive capacity of DU-145 cells. Because of the heterogenous nature of prostatic tumors, we examined promatrilysin expression in cocultures containing two different prostatic carcinoma cell lines, DU-145 and LNCaP. METHODS: Using enzyme linked immunosorbent assay (ELISA) analyses, promatrilysin expression was measured in DU-145/LNCaP cocultures and conditioned media cross cultures. The effects of blocking IL-6 on promatrilysin expression were examined by pretreating conditioned media with IL-6 neutralizing antibody. RESULTS: A significant induction of promatrilysin expression was observed in DU-145/LNCaP cocultures compared to LNCaP cells alone. In addition, DU-145 conditioned medium induced the same fold induction of promatrilysin as was observed in the cocultures. LNCaP cell conditioned medium did not induce promatrilysin expression in DU-145 cells. Neutralization of IL-6 with neutralizing antibody abrogated DU 145 conditioned media induced promatrilysin expression to baseline levels. CONCLUSIONS: IL-6 secreted by DU-145 cells can induce promatrilysin expression in LNCaP cells. IL-6, in vivo, may act as a paracrine signaling factor that regulates matrix metalloproteinase expression. Therefore, IL-6 may play a role in invasive metastatic processes of a prostate carcinoma. PMID- 11494337 TI - Role of canine basal cells in postnatal prostatic development, induction of hyperplasia, and sex hormone-stimulated growth; and the ductal origin of carcinoma. AB - BACKGROUND: The canine prostate has often been proposed as a model for abnormal growth of the human gland. Hyperplasia of the prostate is common in aging men and has been estimated to be present in 100% of old intact dogs. While prostatic carcinoma is common in older men, it appears to be rare in dogs and unlike the disease in humans, it occurs with relatively high frequency in castrated animals. Since basal cells are thought to be key participants in normal and abnormal growth of the human gland, we used immunohistochemistry to investigate the role that they may play in canine prostatic development, the evolution of hyperplasia and carcinoma, and the effects of sex hormones on these cells. METHODS: Prostate specimens were obtained at autopsy from seven sexually immature dogs, autopsy and biopsy samples from 14 sexually mature intact animals, from four castrates, and from19 dogs with prostatic carcinoma. In addition, we also studied the prostates from two intact dogs treated with 5alpha-dihydrotestosterone (DHT) for 6 months and two castrated dogs that were subsequently treated with 5alpha-androstane 3alpha diol and estradiol-17alpha, as well as specimens from two sexually ablated animals given DHT for 2 weeks. All specimens were immunostained for high molecular weight cytokeratin (HMC), pancytokeratin, androgen receptor (AR), and the proliferative marker KI-67. RESULTS: We find that basal cells are the major proliferative cell type in the neonatal and adult canine prostate and that the expression of HMC staining, which defines these cells, may be regulated by androgens. In the adult gland, ductal basal cells formed a contiguous layer, whereas those lining acini were discontinuous. Populations of both basal cell types were variably AR positive, but while HMC immunostaining was abolished in acinar cells following long-term castration, staining remained in ductal cell counterparts. Paralleling the histological development of hyperplasia, the acinar basal cell population increased with age and were the major cell type that expressed KI-67. In contrast, ductal basal cell populations did not expand in the prostates of older dogs and were seldom positively stained for KI-67. The numbers of HMC and KI-67-stained acinar basal cells were dramatically increased in the prostates of intact dogs treated with DHT when compared with glands of untreated controls. This was not the case with ductal basal cells. Androgens given alone or together with estrogen to castrated dogs induced widespread HMC and KI-67 immunostaining in both populations of basal cells. In addition, our results indicate that the majority of canine prostatic carcinomas likely arise exclusively from ductal epithelium. Only one of the 19 cases of carcinoma contained cells that expressed AR, which suggests that androgens may not be required for the initiation or progression of these cancers. CONCLUSIONS: Our findings indicate that two biologically distinct populations of basal cells may exist in the canine prostate. In this regard, the age-related expansion of proliferating acinar basal cell populations, probably mediated by sex steroids, is a key factor in the pathogenesis of canine prostatic hyperplasia. Additionally, we find that prostatic carcinoma in the dog likely arises from ductal cells. Taken together, these findings may indicate that canine acinar basal cells and ductal epithelium have separate susceptibilities to factors that promote hyperplastic or neoplastic development. PMID- 11494338 TI - Benzene and lymphohematopoietic malignancies in humans. AB - BACKGROUND: Quantitative evaluations of benzene-associated risk for cancer have relied primarily on findings from a cohort study of highly exposed U.S. rubber workers. An epidemiologic investigation in China (NCI/CAPM study) extended quantitative evaluations of cancer risk to a broader range of benzene exposures, particularly at lower levels. METHODS: We review the evidence implicating benzene in the etiology of hematopoietic disorders, clarify methodologic aspects of the NCI/CAPM study, and examine the study in the context of the broader literature on health effects associated with occupational benzene exposure. RESULTS: Quantitative relationships for cancer risk from China and the U.S. show a relatively smooth increase in risk for acute myeloid leukemia and related conditions over a broad dose range of benzene exposure (below 200 ppm-years mostly from the China study and above 200 ppm-years mostly from the U.S. study). CONCLUSIONS: Risks of acute myeloid leukemia and other malignant and nonmalignant hematopoietic disorders associated with benzene exposure in China are consistent with other information about benzene exposure, hematotoxicity, and cancer risk, extending evidence for hematopoietic cancer risks to levels substantially lower than had previously been established. Published 2001 Wiley-Liss, Inc. PMID- 11494339 TI - A pilot evaluation of tibia lead concentrations in Taiwan. AB - BACKGROUND: The aims of this study were to examine some of the factors that influence tibia lead concentrations, tibia lead x-ray fluorescence measurement uncertainty and blood lead concentrations, and to compare tibia lead concentrations in Taiwanese lead workers to those observed in lead workers from other countries. METHODS: A pilot evaluation of 43 adult lead workers who underwent measurements of tibia lead and blood lead concentrations. RESULTS: Mean and maximum tibia lead concentrations were 54 microg of Pb per g of bone mineral(microg/g) and 193 microg/g, respectively. Mean and maximum blood lead concentrations were 44 microg/dl and 92 microg/dl, respectively. CONCLUSION: Past occupational control of lead exposure in Taiwan, ROC, did not prevent these workers from accumulating tibia lead concentrations greater than those in similar workers elsewhere in the world. PMID- 11494340 TI - Respiratory effects of exposure to low levels of concrete dust containing crystalline silica. AB - BACKGROUND: Dusts containing crystalline silica are generated in mining, construction, glass, granite and concrete production industries. The association between exposure to low levels of concrete dust containing crystalline silica and reduction in lung function, was evaluated in a cross-sectional study. METHODS: The study was carried out among 144 concrete workers, from two factories, with exposure assessment of respirable dust and silica by personal samplers. Results of respiratory questionnaires and standardized measurements of lung function were compared with the results in a control population. Multiple linear regression analysis was used in selecting factors that predict (age and standing height standardized residual) lung function. RESULTS: The average concentration of respirable dust in both factories was 0.8 mg/m(3) and 0.06 mg/m(3) for respirable silica. The average silica content of the dust was 9%. The average cumulative dust exposure was 7.0 mg/m(3) year and cumulative silica exposure was 0.6 mg/m(3) year. Significant associations between exposure to concrete dust and a small lung function (FEV(1)/FVC ratio, MMEF) loss were found, independent of smoking habits and of a history of allergy. CONCLUSIONS: Our results indicate that, concrete workers with chronic obstructive pulmonary symptoms and/or work-related lower respiratory symptoms are at risk of having a reduction in lung function (FEV&(1)/FVC ratio) outside the 5th percentile of the external reference population, and therefore, of mild chronic obstructive pulmonary disease, at respirable concrete dust levels below 1 mg/m(3) with a respirable crystalline silica content of 10% (TWA, 8 hr). PMID- 11494341 TI - Intervention for prevention of low back pain in Japanese forklift workers. AB - BACKGROUND: The effectiveness of two different approaches for the prevention of low back pain (LBP) was compared in forklift workers. The first approach (personal) consisted of providing lumbar support, arctic jacket and physical exercise, and the second (facility approach) included the improvement of forklift seats and tires. METHODS: The self-reported prevalence of LBP was surveyed three times before and after the two forms of interventions, in 260 male blue-collar workers including 27 forklift workers, and 55 male white-collar workers of a copper smelter. RESULTS: The initial prevalence of LBP was 63% in the forklift workers, which was significantly higher than that found in the other blue-collar workers (32%) and in the white-collar workers (22%). One year after the first intervention (personal approach) to the forklift workers, the prevalence of LBP fell to 56%. The second intervention (facility approach), which was mainly comprised of a reduction in whole body vibration, was subsequently added, and 9 months later the prevalence of LBP in the forklift workers further decreased to 33%. The reduction of the prevalence from the initial survey was significant (P = 0.008), and that from the second survey was nearly significant (P = 0.070). CONCLUSIONS: These findings suggest that the facility approach is more effective for a reduction of LBP than the personal approach. PMID- 11494342 TI - Evaluation of the effectiveness of educational interventions in the Pennsylvania Central Region Farm Safety Pilot Project. AB - BACKGROUND: Evaluation of agricultural safety interventions has frequently been identified as an area requiring further research. This study prospectively evaluates the effectiveness of three specific educational safety interventions in reducing farm hazards. METHODS: Farm characteristics and hazard conditions at 216 farms in Pennsylvania were assessed through a questionnaire and objective audit, respectively, at both pre- and post-intervention time points. Counties were assigned to one of the following interventions: youth education, community coalition, self-audit, pre/post control, or post-only control group. Changes in hazard were analyzed through linear regression. RESULTS: Self-audit was the most effective intervention, leading to a 20% reduction in hazard scores. The community coalition and pre/post control group also showed reductions. CONCLUSIONS: Intervention effectiveness significantly differed depending on initial hazards, indicating the need to target specific interventions for more or less hazardous farms. Findings of this prospective evaluation differed from the initial cross-sectional results, thus underscoring the need for longitudinal investigations. PMID- 11494343 TI - Pesticide exposure beliefs among Latino farmworkers in North Carolina's Christmas tree industry. AB - BACKGROUND: Pesticide exposure is a major preventable occupational hazard for farmworkers. This study examined the beliefs of Latino farmworkers in North Carolina's Christmas tree industry regarding pesticide exposure. METHODS: In depth interviews were conducted with 20 Mexican male seasonal farmworkers. Participants discussed beliefs about agricultural chemicals, routes of exposure, and health effects of these chemicals. They also discussed their knowledge and use of pesticide safety practices and safety training received. RESULTS: Most farmworkers knew that pesticides could be harmful, though workers varied in their levels of knowledge regarding routes of exposure, specific health effects of pesticides, and ways to avoid and reduce exposure. Workers varied considerably in the amount of safety training received and use of safety practices. Perceived lack of control and health beliefs were salient factors that decreased workers' use of safety practices. CONCLUSIONS: This study adds to the growing body of research which documents the health beliefs of Latino farmworkers in the U.S. relative to pesticides and pesticide safety. This literature is beginning to show convergence on several points (e.g., farmworker knowledge of acute vs. long-term illness resulting from pesticide exposure), as well as regional variation in pesticide safety beliefs. This study substantiates the need for pesticide safety education to address issues of control as well as beliefs. PMID- 11494344 TI - Work environment factors, health, lifestyle and marital status as predictors of job change and early retirement in physically heavy occupations. AB - BACKGROUND: The aim was to assess the rate at which waste collectors and municipal workers leave their job, to establish the outcome and to identify associated risk factors within work environment, health, lifestyle and marital status. METHODS: A questionnaire-based survey among a cohort of 2,918 waste collectors and municipal workers was performed in 1994, with follow-up 2.5 years later. RESULTS: Twenty-five percent had left the job at follow-up. Of these, 31% had changed jobs (associated with low decision authority), 16% were unemployed (associated with low skill discretion, pushing heavy loads and extreme bending of back), 10% received Disability Pension/long-term sick leave (associated with low skill discretion, prevalent diseases, underweight and smoking), and 12% received Early Retirement Pension (associated with extreme bending of back and marital status). CONCLUSIONS: The study suggests a potential for preventing people from leaving physically heavy occupations by improving the physical and psychosocial work environment. PMID- 11494345 TI - Health effects among employees in sewage treatment plants: A literature survey. AB - BACKGROUND: Over the years, there have been regular reports in the media of work related symptoms among employees at sewage treatment plants. Concern has spread among employees over the lack of knowledge of risk agents' symptoms, and diseases in that environment. This paper reviews the investigations of health risks in employees working in the sewage treatment plants. METHODS: A literature search was performed with the search terms; sewage, waste water, health effects, infection, and health hazards. Articles on health effects in relation to sewage were selected. RESULTS: Work in sewage water plants can involve exposure to different types of microorganisms and chemicals. The bacterial exposure is dominated by bacteria that naturally occur in nature. However, different bacteria and viruses that give rise to infections can be present in this environment and thus there exists a risk of infection, especially of hepatitis A. Investigations suggest that gastrointestinal tract symptoms are more common among employees at sewage treatment plants than among controls. Respiratory symptoms, fatigue, and headache have also been reported in several investigations. The cause of the symptoms is unknown, although certain data suggest that they are caused by inflammation. The results suggest that endotoxin in Gram-negative bacteria may be one of the causative agents. As regards cancer, some studies report an increased risk of stomach cancer and a few studies report an increased risk of cancer in the larynx, liver or, prostate or of leukemia. The spread of the cancers over a multitude of organs does not support a hypothesis of causality with agents commonly found in sewage treatment plants. CONCLUSIONS: Further investigations are needed to determine the work-related effects and ascertain the causal agents. PMID- 11494346 TI - Observed versus predicted carboxyhemoglobin levels in cellulose triacetate workers exposed to methylene chloride. AB - BACKGROUND: Occupational exposure to methylene chloride, together with carboxyhemoglobin concentrations, has not been studied previously. METHODS: Carboxyhemoglobin levels were measured in non-smoking employees exposed to varying concentrations of methylene chloride during the manufacture of cellulose triacetate fibers. The observed carboxyhemoglobin levels were compared to predicted concentrations using a pharmacokinetic model. RESULTS: The presence of carboxyhemoglobin in non-smokers exposed to methylene chloride results primarily from the metabolism of methylene chloride in the liver and exhibits a linear dose response relationship. The observed levels of carboxyhemoglobin in non-smokers at the end of an 8-hour shift depend upon exposures to methylene chloride that day but are independent of occupational exposures on previous days. The observed daily concentrations of carboxyhemoglobin are consistent with predicted concentrations using a pharmacokinetic model. While varying exposure patterns were shown to change the rate of metabolite formation at the end of shift, these same exposure patterns had almost no effect on the total amount of carbon monoxide in the blood. CONCLUSION: While the present study addresses the relationship between methylene chloride, carbon monoxide, carboxyhemoglobin and ischemic heart disease, it does not address the issue of tumorigenicity, which is also the basis for the current U.S. Occupational Health and Safety workplace exposure limit of 25 ppm. This study provides support for the conclusion that the current American Conference of Governmental Industrial Hygienists 8-hour Threshold Limit Value of 50 ppm adequately protects human health with regard to ischemic heart disease and carboxyhemoglobin formation among non-smokers. PMID- 11494347 TI - Prevalence of neuropsychiatric and mucous membrane irritation complaints among Palestinian shoe factory workers exposed to organic solvents and plastic compounds. AB - BACKGROUND: Long-term exposure to organic solvents is associated with neuropsychiatric and mucus membrane irritation. In developing countries, efforts to secure a good working environment are inadequate and protection against chemical exposures is often neglected. METHODS: In a cross-sectional survey, the prevalence of self-reported health complaints and the association with long-term exposure to solvents and plastic materials in the work environment was studied among 167 shoe-factory workers. Prevalences and adjusted prevalence ratios (PR)were calculated in Cox regression. 95% confidence intervals were estimated for PRs of adverse health effects. RESULTS: Overall, the workers reported high prevalences of neuropsychiatric and mucus membrane complaints: headache (65%), mental irritability (53%), tingling of limbs (46%), and sore eyes (43%). Cleaning work was associated with tingling of limbs (PR = 1.8, 1.0-3.2), sore eyes (PR = 1.9, 1.1-3.3), and breathing difficulty (PR = 2.0, 1.0-3.9); plastic work was associated with tingling of limbs (PR = 1.8, 1.2-2.9)and sore eyes (PR = 1.7, 1.1 2.7); and varnishing was associated with breathing difficulty (PR = 1.9, 1.1 3.5). CONCLUSIONS: The high prevalence of self-reported health complaints and the exposure-outcome association could be due to volatile organic solvents (dichloromethane, n-hexane)and plastic compounds (isocyanates and polyvinyl chloride). Absence of a satisfactory work environment is likely to contribute to high exposure levels. PMID- 11494348 TI - Medical aspects of global warming. AB - BACKGROUND: Global warming is caused by increased carbon dioxide (CO2)resulting in a greenhouse effect with enhanced warming of the earth. Measurements of CO2 show a steady increase over the past 30 years caused by the burning of fossil fuels and from the loss of natural CO2 sinks. A 100-year increase in global temperature by 0.3 to 0.6 degrees C is reflected in atmospheric warming, glacier shrinkage, and rising sea levels. OBJECTIVES: Planetary ecosystem dynamics are being altered, challenging public health. It is predicted that morbidity and mortality will increase as a result of heat stress, as seen in recent heat waves in the U.S. Weather disaster effects will increase in number and magnitude, and both noninfectious and infectious diseases may flourish. A significant challenge will be the changes in life cycles of microbial species due to the warmer environs. Specific increases in incidence have been noted for vector-borne diseases, in addition to pulmonary findings, cardiovascular morbidity, neurological diseases, and occupational diseases. CONCLUSIONS: Warming can be demonstrated by the observed changes that have already occurred in the environment, particularly the thinning of polar ice caps. The United States Global Research Program has been established to coordinate research activities, which responds to issues deemed important by the United Nations Framework Convention on Climate Change. Research issues pertain to the scientific uncertainties in the greenhouse effect, temperature measurements at various atmospheric levels and latitudes, and impact on biota redistribution. The Kyoto Protocol has mandated specific solutions, e.g., a 7% reduction in CO2 levels within 10 years. Future recommendations involve supporting new technologies that are available to decrease emissions as well as understanding the role that occupational and environmental specialists have in global warming recognition. PMID- 11494349 TI - Malignant mesothelioma from neighborhood exposure to anthophyllite asbestos. AB - BACKGROUND: Anthophyllite asbestos has been reported to cause asbestosis, lung cancer, mesothelioma, and pleural plaques in occupationally exposed workers. Anthophyllite has also been associated with pleural plaques in Finland and Japan among those who live near mines and mills and have neighborhood or environmental exposure. METHODS: We evaluated a 38-year-old patient with pleural mesothelioma who lived, attended school, and delivered newspapers near a manufacturing facility that used exclusively anthophyllite asbestos fiber from ages 8-17 years. He had no work exposure to asbestos. RESULTS: The pleural mesothelioma was an epithelial type with tubulopapillary structures and was treated with an extrapleural pneumonectomy followed by radiation therapy. The malignant cells were positive by immunohistochemistry for cytokeratin but negative for carcinoembryonic antigen, S100, B72.3, and leu M1 antigen. Anthophyllite fibers were > 5 microm in length in lung tissue compared to 3 microm from a general population study. CONCLUSIONS: Anthophyllite asbestos has been associated with neighborhood environmental exposure and pleural plaques; we now report a neighborhood exposure and pleural mesothelioma. PMID- 11494350 TI - Fatal unintentional occupational poisonings by hydrofluoric acid in the U.S. AB - BACKGROUND: Case reports have identified hydrofluoric acid (HF) as causing fatal work injury, and HF has both local and systemic toxicity. Surveillance for HF related mortality is problematic because of the lack of unique coding for this acid in hospital records and vital statistics. METHODS: We identified HF-related fatal work injuries investigated by the Occupational Safety and Health Administration (OSHA) for 1984-94 from coding of Hazardous Substance 1460 (HF) and requested case investigation files under the Freedom of Information Act. We attempted to identify HF-related deaths in the US for the same period through literature case reports, the Consumer Product Safety Commission, and the American Association of Poison Control Centers (AAPCC). RESULTS: For the 11 year period, OSHA investigated nine deaths in eight incidents which involved HF. Four deaths were from skin contact with concentrated HF, and five deaths involved both skin contact and inhalation of vapor. Unsafe work practices were factors in all of the deaths. Calcium chloride or gluconate was noted to have been administered to five of the nine victims. Calcium was administered 90 min after exposure to two victims, and more than 6 h after exposure to a third. We were able to establish that the regional poison control center had been contacted in regard to only one victim. For the period 1984-94, we were able to identify no additional deaths from CPSC reports, one additional death from AAPCC annual reports, and four other deaths from case reports in the medical literature. CONCLUSIONS: For the period of this study, OSHA records identified the greatest number of HF-related fatalities. The limited information in the records suggest that some victims did not receive appropriate medical care, nor was the regional poison center contacted regarding care. The full extent of health problems related to HF could be better quantified if usual surveillance sources, such as vital records, included unique coding for this acid. PMID- 11494351 TI - Interstitial fibrosis following hydrogen sulfide exposure. AB - This case report describes a patient with interstitial fibrosis 4 years after poisoning by hydrogen sulfide. Hydrogen sulfide causes pulmonary edema and is also toxic to the nervous system. Long-term pulmonary sequelae of hydrogen sulfide poisoning has not been reported frequently in the literature. PMID- 11494353 TI - Neurofibromatosis type 1: II. Answers from animal models. PMID- 11494354 TI - Direct cell-cell interactions control apoptosis and oligodendrocyte marker expression of neuroepithelial cells. AB - During brain development, the neuroepithelium generates neurons and glial cells. Proliferation and differentiation of neuroepithelial cells are controlled by a complex combination of secreted factors and more intrinsic or local mechanisms, such as lateral inhibition and asymmetric division. To obtain further insights into the signals governing neuroepithelial cell fate, we used the immortomouse to derive conditionally immortalised cell lines from mouse E10 neuroepithelium. We isolated a nestin-positive basic fibroblast growth factor (bFGF)-responsive cell line (SVE10-23) which mostly differentiate into astrocytes when cocultured with primary cortical cells. We found that, by simply lowering the cell density, SVE10 23 cells embarked on oligodendrocytic differentiation as indicated by the strong expression of galactocerebroside C and 2'3'-cyclic nucleotide 3' phosphodiesterase. Apoptosis accompanied the differentiation, and all cells died within 1 week. We present here evidence that direct interactions between cells are the main mechanism regulating this oligodendrocytic differentiation. We demonstrate that SVE10-23 cells contact or proximity inhibit their differentiation, prevent apoptosis, and promote their proliferation. Similarly, others nestin-positive precursor cell lines and nonimmortalised bFGF-grown E10 cells were found to spontaneously differentiate at low density, thus generalising the idea that neural precursor fate is regulated by direct cell-cell interactions. The SVE10-23 cell line provides a valuable tool with which to study further the molecular components implicated in this mode of regulation. PMID- 11494355 TI - Efficient gene transfer in mouse neural precursors with a bicistronic retroviral vector. AB - Gene transfer into neural precursors is a powerful approach to study the function of specific gene products during nervous system development. Here we describe a retrovirus-based methodology to transduce foreign genes into mouse neural precursors. We used a high-titer bicistronic retroviral vector that encodes a marker gene, placental alkaline phosphatase (plap), and a selection gene, neomycin phosphotransferase II (neoR), under the translational control of two retroviral internal ribosome entry segments. Transduction efficiency even without selection was up to 95% for multipotential neurospheres derived from embryonic striata and grown with basic fibroblast growth factor 2. Expression of plap and neoR was sustained with time in culture and upon differentiation into neurons, astrocytes, and oligodendrocytes, as shown by double immunofluorescence labeling with cell type-specific markers, Western blotting, and neomycin resistance. However, levels of plap were decreased in differentiated oligodendrocytes. Transduction with the same vector of neonatal oligodendrocyte precursors grown in oligospheres consistently resulted in a lower proportion of plap-immunoreactive cells and enhanced cell death in the absence of neomycin. However, plap expression was maintained in some differentiated oligodendrocytes expressing galactocerebroside or myelin basic protein. In that neurospheres can be easily expanded in vitro and factors enabling their differentiation into the three main central nervous system cell types are being elucidated, this methodology could be used in the future to produce large number of transduced, differentiated neural cells. PMID- 11494356 TI - Direct isolation of committed neuronal progenitor cells from transgenic mice coexpressing spectrally distinct fluorescent proteins regulated by stage-specific neural promoters. AB - Many tissues arise from pluripotent stem cells through cell-type specification and maturation. In the bone marrow, primitive stem cells generate all the different types of blood cells via the sequential differentiation of increasingly committed progenitor cells. Cell-surface markers that clearly distinguish stem cells, restricted progenitors, and differentiated progeny have enabled researchers to isolate these cells and to study the regulatory mechanisms of hematopoiesis. Neuronal differentiation appears to involve similar mechanisms. However, neural progenitor cells that are restricted to a neuronal fate have not been characterized in vivo, because specific cell-surface markers are not available. We have developed an alternative strategy to identify and isolate neuronal progenitor cells based on dual-color fluorescent proteins. To identify and isolate directly progenitor cells from brain tissue without the need for either transfection or intervening cell culture, we established lines of transgenic mice bearing fluorescent transgenes regulated by neural promoters. One set of transgenic lines expressed enhanced yellow fluorescent protein (EYFP) in neuronal progenitor cells and neurons under the control of the Talpha1 alpha tubulin promoter. Another line expressed enhanced green fluorescent protein (EGFP) in immature neural cells under the control of the enhancer/promoter elements of the nestin gene. By crossing these lines we obtained mice expressing both transgenes. To isolate neuronal progenitor cells directly from the developing brain, we used flow cytometry, selecting cells that expressed EGFP and EYFP simultaneously. We expect this strategy to provide valuable material with which to study the mechanisms of neurogenesis and to develop cell-based therapies for neurological disorders. PMID- 11494357 TI - Administration of FGF-2 to embryonic mouse brain induces hydrocephalic brain morphology and aberrant differentiation of neurons in the postnatal cerebral cortex. AB - Fibroblast growth factor-2 (FGF-2) was injected into mouse cerebral ventricles at embryonic day (E) 14 in utero and its effects on developing brain morphology and expression of various cell- or differentiation-associated protein markers in the cerebral cortex were examined. High doses of FGF-2 (200 or 300 ng) caused encephalic alternations such as deformation of the calvarium, enlargement of the ventricular spaces, and thinning of the cerebral cortex. There was no gross abnormality in the alignment of the cerebral neuronal layers, however, both cell number and cell density of the upper layers (II/III) and the lower layers (IV-VI) of the cerebral cortex were increased. Brain-derived neurotrophic factor (BDNF), tyrosine hydroxylase, nestin, and microtubule-associated protein 2 were aberrantly or ectopically expressed in the deep areas of the cerebral cortex. A substantial number of these cells coexpressed these antigens. These observations demonstrate that a subpopulation of neurons in the cortical deep layer abnormally differentiated or partly sustained their immature state following a single administration of FGF-2 at E14. Developmental analysis of localization of BDNF positive cells suggested that the abnormality started around P5. Furthermore, cell migration was not affected by FGF-2 administration. FGF-2 seems to play predominant roles in the proliferation of neuronal precursors and in neuronal differentiation in the developing mouse cerebral cortex even at relatively late stages of brain neurogenesis. PMID- 11494358 TI - Movement of endoplasmic reticulum in the living axon is distinct from other membranous vesicles in its rate, form, and sensitivity to microtubule inhibitors. AB - The endoplasmic reticulum (ER) is the major membranous component present throughout the axon. Although other membranous structures such as synaptic vesicles are known to move via fast axonal transport, the dynamics of ER in the axon still remains unknown. To study the dynamics of ER in the axon, we have directly visualized the movement of two ER-specific membrane proteins, the sarcoplasmic/endoplasmic reticulum calcium-ATPase and the inositol 1,4,5 trisphosphate receptor, both of which were tagged with green fluorescence protein (GFP) and expressed in cultured chick dorsal root ganglion neurons. In contrast to GFP-tagged synaptophysin that moved as vesicles at 1 microm/sec predominantly in the anterograde direction in the typical style of fast axonal transport, the two ER proteins did not move in a discrete vesicular form. Their movement determined by the fluorescence recovery after photobleaching technique was bi directional, 10-fold slower (approximately 0.1 microm/sec), and temperature sensitive. The rate of movement of ER was also sensitive to low doses of vinblastine and nocodazole that did not affect the rate of synaptophysin-GFP, further suggesting that it is also distinct from the well-documented movement of membranous vesicles in its relation with microtubules. PMID- 11494359 TI - Modulation of HSP25 expression during anterior horn motor neuron degeneration in the paralyse mouse mutant. AB - The paralyse spontaneous mutation in mice involves degeneration and death of anterior horn motor neurons. Mutant mice are not viable past postnatal day 16. At present, the mechanisms involved in motor neuron death are unknown. Here, we investigate the expression of the small heat shock protein Hsp25, in the spinal cord of paralyse at two different stages during postnatal development, i.e., day 11 and day 14. Western blot analysis reveals that the level of Hsp25 was strikingly different in paralyse as compared to control littermates. Hsp25 expression level in paralyse at day 11 was much lower than in control mice. At day 14, an opposite pattern was observed. Such pattern seems to be restricted to spinal cord, since level of Hsp25 in other tissues (lung, brain, liver, and heart) was quite similar. Immunofluorescence examination of the lumbar spinal cord sections reveals that in control mice, Hsp25 was expressed at high level in motor neurons located in the ventral horn at both day 11 and day 14. By contrast, in paralyse mice, Hsp25 staining within the motor neurons was barely detectable except as a spot in the nucleolus (day 11). At the end stage of the disease (day 14), not only was Hsp25 staining even less intense in motor neurons, but also a strong Hsp25 staining was observed in reactive astrocytes within the gray matter. Taken together, these data suggest that Hsp25 expression is differently modulated in neuronal and glial cells during neurodegenerative processes leading to motor neuron death. PMID- 11494360 TI - Tyrosine hydroxylase and dopamine transporter expression in residual dopaminergic neurons: potential contributors to spontaneous recovery from experimental Parkinsonism. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahyrdropyridine (MPTP)-exposed cats develop severe Parkinsonism that spontaneously resolves in 4-6 weeks. The present study examined the extent to which compensatory changes in tyrosine hydroxylase (TH) and dopamine transporter (DAT) gene and protein expression may underlie this behavioral recovery. In normal cats, TH and DAT protein levels were higher in the dorsal vs. ventral striatum. Expression of DAT and TH mRNA was higher in substantia nigra pars compacta (SNc) than in the ventral tegmental area (VTA). In symptomatic parkinsonian animals, DAT and TH protein levels were significantly decreased in all striatal areas studied. TH and DAT mRNA expression in residual SNc neurons were decreased a mean 32% and 38%, respectively. DAT gene expression in residual VTA neurons in symptomatic animals was decreased 30% whereas TH gene expression was unaffected. In spontaneously recovered cats, TH protein levels were significantly higher than the levels in symptomatic cats only in the ventral striatum, whereas no increase in DAT protein levels were observed in any striatal area. Residual neurons in most ventral mesencephalic regions of recovered cats had increased TH mRNA expression but not increased DAT gene expression, compared with symptomatic animals. Thus, increased TH protein and mRNA and suppression of DAT protein and mRNA expression in the striatum and ventral mesencephalon were associated with functional recovery from MPTP-induced parkinsonism. PMID- 11494361 TI - Role of GluR2 expression in AMPA-induced toxicity in cultured murine cerebral cortical neurons. AB - alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPA-R) mediated neurotoxicity was studied in relation to subunit expression and the presence of Ca(2+)-permeable receptor channels. AMPA-mediated toxicity had two components: 1) a direct AMPA-R-mediated component, which was not due to Ca(2+) influx through voltage-gated Ca(2+) channels, reversal of the Na(+)/Ca(2+) exchanger or release of calcium from dantrolene-sensitive intracellular Ca(2+) stores, and 2) a minor, indirect component involving activation of NMDA receptor channels, because of glutamate release and removal of the Mg(2+) block of the NMDA receptor on AMPA-R stimulation. The involvement of Ca(2+) influx through AMPA-R was also examined. The number of neurons possessing Ca(2+)-permeable AMPA R increased during culture development, concurrently with an increasing susceptibility for AMPA-induced toxicity during development. GluR2(R) levels also increased during development, and channel blockers of Ca(2+)-permeable AMPA-R lacking the GluR2(R) subunit (spermine and philanthotoxin) failed to prevent neurotoxicity or increases in [Ca(2+)](i). Thus, the direct AMPA-R-mediated toxicity may be explained by initiation of cell death by Ca(2+) fluxing through AMPA-R containing GluR2(R). The components of direct AMPA-R-mediated toxicity are proposed to be 1) toxicity mediated by GluR2(R)-lacking AMPA-R and 2) toxicity mediated by low-Ca(2+)-permeability AMPA-R containing GluR2(R). PMID- 11494362 TI - Models of brain injury and alterations in synaptic plasticity. AB - Animal models are crucial for understanding human pathophysiological processes and for understanding how connections are injured, lost, or even regenerated and/or repaired. When animal models are used in conjunction with theoretical computational models, an ideal combination is achieved that potentially yields insight and encourages the formation of new theories concerning connectionism, cognitive functioning, and synaptic mechanisms. Mechanisms regulating glutamate receptor activation and intracellular calcium levels are important for normal synaptic transmission. These mechanisms (and others) are also critical during and after brain injury when the potential exists for these mechanisms to function pathologically. Interestingly enough, the regulation of glutamate receptor activation and intracellular calcium levels is also involved in normal processes of neuronal and synaptic plasticity. In addition, studies have shown that neurotrophins and cytokines, which are released after brain injury, can be neuroprotective and may also be important in synaptic plasticity. Furthermore, synaptic plasticity is a phenomenon thought by many to be necessary for memory encoding. If this is the case, then research described in this review has significant scientific merit concerning plasticity and memory and clinical benefit for understanding pathophysiologic processes associated with brain injury and memory impairment. This paper reviews the application of experimental animal models of brain injury for simulating conditions of stroke, trauma, and epilepsy (and/or seizure generation) and the associated cellular mechanisms of brain injury. The paper also briefly addresses the advantage of using computational models in combination with experimental models for hypothesis building and for aiding in the interpretation of empirical data. Finally, it reviews studies concerning brain injury and synaptic plasticity. PMID- 11494363 TI - In vitro generation and transplantation of precursor-derived human dopamine neurons. AB - The use of in vitro expanded human CNS precursors has the potential to overcome some of the ethical, logistic and technical problems of fetal tissue transplantation in Parkinson disease. Cultured rat mesencephalic precursors proliferate in response to bFGF and upon mitogen withdrawal, differentiate into functional dopamine neurons that alleviate motor symptoms in Parkinsonian rats (Studer et al. [1998] Nat. Neurosci. 1:290-295). The successful clinical application of CNS precursor technology in Parkinson disease will depend on the efficient in vitro generation of human dopaminergic neurons. We demonstrate that human dopamine neurons can be generated from both midbrain and cortical precursors. Transplantation of midbrain precursor-derived dopamine neurons into Parkinsonian rats resulted in grafts rich in tyrosine hydroxylase positive neurons 6 weeks after transplantation. No surviving tyrosine hydroxylase positive neurons could be detected when dopamine neurons derived from cortical precursors were grafted. Our data demonstrate in vitro derivation of human dopamine neurons from expanded CNS precursors and encourage further studies that systematically address in vivo function and clinical potential. PMID- 11494364 TI - Age-dependent and tissue-specific CAG repeat instability occurs in mouse knock-in for a mutant Huntington's disease gene. AB - Huntington's disease (HD) is a neurodegenerative disorder characterized by the expansion of CAG repeats in exon 1 of the HD gene. To clarify the instability of expanded CAG repeats in HD patients, an HD model mouse has been generated by gene replacement with human exon 1 of the HD gene with expansion to 77 CAG repeats. Chimeric proteins composed of human mutated exon 1 and mouse huntingtin are expressed ubiquitously in brain and peripheral tissues. One or two CAG repeat expansion was found in litters from paternal transmission, whereas contraction of CAG repeat in litters was observed through maternal transmission. Elderly mice show greater CAG repeat instability than younger mice, and a unique case was observed of an expanded 97 CAG repeat mouse. Somatic CAG repeat instability is particularly pronounced in the liver, kidney, stomach, and brain but not in the cerebellum of 100-week-old mice. The same results of expanded CAG repeat instability as observed in this HD model mouse were confirmed in the human brain of HD patients. Glial fibrillary acidic protein (GFAP)-positive cells have been found to be increased in the substantia nigra (SN), globus pallidus (GP), and striatum (St) in the brains of 40-week-old affected mice, although without neuronal cell death. The CAG repeat instability and increase in GFAP-positive cells in this mouse model appear to mirror the abnormalities in HD patients. The HD model mouse may therefore have advantages for investigations of molecular mechanisms underlying instability of CAG repeats. PMID- 11494365 TI - IL-6 deficiency allows for enhanced therapeutic value after bone marrow transplantation across a minor histocompatibility barrier in the twitcher (globoid cell leukodystrophy) mouse. AB - Bone marrow transplantation (BMT) has therapeutic value for twitcher (globoid cell leukodystrophy) mice, which suffer from a genetic deficiency of the lysosomal enzyme galactosylceramidase that leads to progressive demyelination and early death. Preliminary investigations indicated that a semiallogeneic BMT resulted in graft vs. host disease (GVHD) in twitcher mice but not normal mice. Increased production of the cytokine IL-6 has been demonstrated in twitcher mice, and it has been linked with induction of GVHD. We investigated the effects of BMT in twitcher/IL-6 deficient mice and compared these findings with those from transplanted twitcher and control mice. After a semiallogeneic BMT, 11.4% of controls died within few weeks while the rest survived >100 days without GVHD. In contrast, 85% of the transplanted twitcher mice died by 70 days and 65% developed clinical signs of GVHD, e.g., alopecia and weight loss. In transplanted twitcher/IL-6 deficient mice, only 21% died by Day 70, none had alopecia, and 23% had weight loss. There was no difference in the onset day and severity of twitching between twitcher and twitcher/IL-6 deficient mice after BMT. In transplanted twitcher/IL-6 deficient mice, there was improvement of BBB integrity and a decrease in globoid cell number compared with nontransplanted twitcher/IL-6 deficient mice. In summary, these results demonstrate that an underlying pathology like globoid cell leukodystrophy leads to activation of GVHD responses in a donor-host combination that would not normally induce GVHD. Furthermore, IL 6 seems to play a key role because a deficiency of IL-6 results in a better prognosis. PMID- 11494366 TI - Proliferating oligodendrocytes are present in both active and chronic inactive multiple sclerosis plaques. AB - The proliferation marker Ki-67 labels cell nuclei in the G(1), S, M, and G(2) phases of the cell cycle. We used Ki-67 immunohistochemistry to quantify proliferating glial cells in brain tissue sections from twenty-four patients, comprised of multiple sclerosis, normal brains, and other neurological disease controls. Glial proliferation was greatly increased in MS lesions when compared with control brain white matter. Both actively demyelinating/early remyelinating plaques and chronic inactive plaques of long standing often displayed large numbers of glial cells in the proliferative cycle. The bulk of these proliferating cells were of oligodendroglial lineage in the MS plaques. Ki-67 positive macrophage/microglial lineage cells were largely restricted to acute lesions. The finding of increased numbers of proliferating oligodendroglia in most MS plaques, regardless of disease duration or activity state, indicates that the MS brain is capable of recruiting unexpectedly large numbers of new oligodendrocytes over long periods of time. The factors within the MS plaque microenvironment that provoke new oligodendrocyte generation and their subsequent loss still need to be identified. PMID- 11494367 TI - Variable galactocerebroside expression by human Schwann cells in dissociated and peripheral nerve explant cultures. AB - It has been well established that rat Schwann cells down regulate their cell surface expression of galactocerebroside (GalC) in vitro under normal cell culture conditions. To determine whether human Schwann cells exhibit a similar down-regulation of GalC in vitro we examined GalC expression in dissociated human Schwann cell cultures derived from normal adult peripheral nerve. Twenty-four hours post-dissociation up to 63% of human Schwann cells were found to express detectable levels of GalC on their surface whereas less than 8% of the Schwann cells expressed detectable levels of GalC at 14 days post-dissociation. In contrast, after nearly 3 months of peripheral nerve explant culture, greater than 30% of human Schwann cells still retained their GalC expression. A similar pattern was also observed when analyzing Schwann cell purity with dissociated cultures exhibiting a rapid decrease in Schwann cell purity under normal culturing conditions although Schwann cell purity was found to be largely unaffected during the period of peripheral nerve explant culture. In summary, we found there was less variation in both GalC expression and Schwann cell purity with time in peripheral nerve explant cultures than dissociated cultures. PMID- 11494368 TI - Neurotrophin secretion from cultured microglia. AB - Because microglia have been suggested to produce neurotrophins, we tested this ability in vitro. Rat primary microglia were found to constitutively secrete a limited amount of brain-derived neurotrophic factor (BDNF), but nerve growth factor (NGF) and neurotrophin-3 (NT-3) were undetectable in the conditioned medium. Stimulation of the cells with lipopolysaccharide (LPS) increased BDNF secretion, and induced NGF secretion. As a first step to examine this regulation system, the association of protein kinase C (PKC) was pharmacologically analyzed. A PKC activator, phorbol-12-myristate-13-acetate, enhanced the secretion of BDNF. Pre-treatment of microglia with a PKC inhibitor, bisindolylmaleimide, suppressed LPS-stimulated BDNF secretion as well as the constitutive one. These results suggest that the PKC signaling cascade is closely associated with BDNF secretion. Among PKC isoforms, PKCalpha probably plays a role in BDNF secretion, based on the results of experiments using a specific PKC activator, 1-oleoyl-2-acetyl-sn glycerol, and a specific PKC inhibitor, Go 6976, and by immunoblotting. Taken together, these findings suggest that the secretion of BDNF from microglia is regulated through PKCalpha-associated signal transduction mechanism. PMID- 11494369 TI - Induction of the proinflammatory cytokine interleukin-18 by axonal injury. AB - Interleukin-18 (IL-18) is an important cytokine in innate immunity and in the induction phase of autoimmunity. We report the expression of IL-18 mRNA and protein after nerve crush during Wallerian degeneration (WD) of the rat nervous system. In normal optic nerves (ON) constitutive IL-18 mRNA levels as revealed by semiquantitative reverse transcriptase polymerase chain reaction were higher than in sciatic nerves (SN). After nerve crush, steady-state levels moderately increased in the distal nerve part of the SN but not the ON. By immunocytochemistry no SN or faint ON IL-18 protein expression was detectable in normal nerves. In contrast, IL-18 expression dramatically increased after SN and ON crush. On the cellular level, ED1(+) macrophages infiltrating the crush site strongly expressed IL-18 at days 2 and 4 after SN crush. By days 4 and 8, in addition, the entire distal nerve part was covered by IL-18(+) macrophages. At day 16, IL-18 immunoreactivity had disappeared despite the persistence of large numbers of ED1(+) macrophages. A similar infiltration of IL-18(+) macrophages was seen at the crush site in the ON. Moreover, microglia in the distal ON stump lacking macrophage infiltration and undergoing delayed myelin degradation up regulated IL-18. In conclusion this study shows that IL-18 is involved in the cytokine network associated with the robust inflammatory response during WD of the SN. Despite up-regulation of the proinflammatory cytokine IL-18, major histocompatibility complex class II, and CD4 molecules similar to macrophages in the PNS, microglial activation after ON injury appears to be insufficient to mount an effective phagocytic response as a prerequisite for successful regeneration in the CNS. PMID- 11494370 TI - Regulation of pp60(c-src) synthesis in rat hippocampal slices by in vitro ischemia and glucocorticoid administration. AB - Corticosteroids, released from the adrenal gland in response to stress, bind to receptors that act as transcription factors to alter gene expression and, subsequently, protein synthesis. Using [(35)S]-methionine-cysteine incorporation to measure protein synthesis in hippocampal slices incubated under ischemic conditions, synthesis of 60 kDa and 78 kDa proteins decreases 4 hr after in vivo administration of corticosterone to rats. The former protein has been identified by immunoblotting and immunoprecipitation to be the proto-oncogene, pp60(c-src). In the absence of prior glucocorticoid administration, ischemic conditions increase the amount of immunoreactive pp60(c-src) in membranes of hippocampal slices. Chronic exposure to elevated titers of glucocorticoids has been demonstrated to result in cell loss as well as in reduced neuronal plasticity and regeneration. Given the involvement of pp60(c-src) in synaptic plasticity and cell growth, glucocorticoid-mediated reduction in its synthesis is a potential molecular marker for stress-induced alterations in brain function. PMID- 11494371 TI - CB1 cannabinoid receptor-mediated neurite remodeling in mouse neuroblastoma N1E 115 cells. AB - The morphological remodeling of neuronal cells influences neurogenesis and brain functions. We hypothesize that psychoactive and neurotoxic effects of cannabinoids may be mediated, at least in part, by their morphoregulatory activities. In the present study, mouse neuroblastoma N1E-115 cells were used as an in vitro model to investigate cannabinoid-induced neurite remodeling effects and to identify the involvement of cannabinoid receptors in this neurite remodeling process. Using reverse transcription-polymerase chain reaction and immunofluorescence microscopy, the endogenously expressed CB1, but not CB2, cannabinoid receptors were detected in morphologically differentiated N1E-115 cells. Activation of these natively expressed CB1 cannabinoid receptors by cannabinoid agonist HU-210 led to a concentration-dependent inhibition of adenylate cyclase activity. Importantly, HU-210 treatment induced neurite retraction in a concentration-dependent manner. Pretreatment of N1E-115 cells with a CB1 antisense oligodeoxynucleotide (ODN) suppressed HU-210-induced inhibition of forskolin-stimulated cAMP accumulation, indicating that the knocking down of functional CB1 cannabinoid receptor expression was achieved. Antisense ODN pretreatment also abolished HU-210-induced neurite retraction, demonstrating the involvement of CB1 cannabinoid receptors in mediating the neurite remodeling effects of HU-210. In addition, reversing HU-210-induced intracellular cAMP declination by 8-Br-cAMP partially prevented HU-210-induced neurite retraction, indicating the involvement of cAMP-dependent signaling pathways in mediating the neurite remodeling function of CB1 cannabinoid receptors in N1E-115 cells. These data demonstrate that neurite remodeling is a newly discovered function of CB1 cannabinoid receptors. This morphoregulatory function of CB1 cannabinoid receptors might be a new mechanism that mediates the psychoactive and neurotoxic effects of cannabinoids in developing and adult brain. PMID- 11494372 TI - Ethanol exposure inhibits the cytotoxic effect induced by gp120 in CHP100 human neuroblastoma cells. AB - The aim of present study was to investigate the acute effects of ethanol on cytotoxicity induced by HIV-1 coat protein gp120 in CHP100 human neuroblastoma cell line. We demonstrate that ethanol, within a range of clinically relevant concentrations (15-90 mM) prevents cell death elicited by gp120 (10 pM) in a dose dependent manner. This protective action seems to be mediated by a reduction of free intracellular Ca(2+) levels because ethanol, at concentrations ranging from 0.1-0.5%, is able to decrease gp120-stimulated Ca(2+) uptake up to 24%. Furthermore, our data show an involvement of NO/cGMP messenger system pathway, because ethanol is also able to reduce gp120-stimulated NO release (up to 45%) and cyclic GMP accumulation (up to 73%). These findings suggest that the protective effect of ethanol against gp120-induced cytotoxicity in CHP100 cells underlies a Ca(2+)-activated, NO/cGMP-dependent mechanism. PMID- 11494374 TI - Experimental research and surgery: why, how, and when? AB - Experimental research faces two great problems: the significant reduction of public funding and the firm opposition of the public opinion. The law forbids the use of large animals, so that it is possible to use small animals only, which require microsurgical techniques. However, even a skillful surgeon does not know how to perform microsurgery and has to begin a long and tiring training to master techniques. We think that experimental surgery should play a role because it tests the validity and safety of new surgical techniques and allows special pathophysiological aspects to be studied. Furthermore experimental surgery could represent an essential stage in the training of young surgeons. We should find a balance between observance of the law and respect of the animals and, on the other hand, the role of experimental surgery because we should not forget that its most important aim is the improvement of the health of the humankind. PMID- 11494375 TI - Basic teaching in microsurgery. AB - We summarize our 15 years of educational experience in the field of teaching microsurgery. The students can be divided into three groups: (1) medical students, (2) researchers, (3) medical doctors and specialists. Characteristics of our method include the following: activity, synchronism, video-assistance, self-controlling, individualization, analysis. The Furka microsurgical educational method, named after one of the authors, is 20 hours long (five 4-hour sessions). The first lesson allows students to become acquainted with the microsurgical instruments. The next lesson consists of learning the probe of layer-feeling. The third lesson is to learn how to produce stitches under the microscope. The fourth lesson includes arterial anastomosis preparation on fresh arterial pieces of animal origin. The fifth lesson means a quality change from previous classes, as practice is performed on living animals, generally rats. The teaching of microsurgical basics requires both patience and empathy. The teaching process is most successful if one teacher deals with a maximum of two students. PMID- 11494376 TI - A new vascularized adrenal transplantation model in the rat. AB - A new microsurgical model of adrenal transplantation in the rat is described. An adrenal graft with its vascular supply, adrenal artery and vein, and the attachment of a segment of aorta and inferior vena cava (IVC) was transplanted to a recipient rat with end-to-side anastomoses between the donor IVC segment and the recipient IVC and between the donor aortic segment and the recipient aorta using 10-0 nylon sutures. Using this model, different groups of recipient rats received iso- or allograft with or without immunosuppressive treatment were tested. This model provides a reliable and useful tool for research on endocrinology. PMID- 11494377 TI - Experimental anaesthesiologic protocol for porto-intracaval shunt for liver total vascular exclusion: preliminary study in the rabbit. AB - The aim of this study was to evaluate the effects and interference of different anaesthetic techniques in rabbits undergoing liver total vascular exclusion using a porto-intracaval shunt. Twenty New Zealand rabbits were divided in three groups: group A receiving diazepam as premedication, ketamine + atropine for induction and maintenance of anaesthesia and undergoing a porto-intracaval shunt operation; group B receiving midazolam as premedication, ketamine + fentanyl + atropine for induction and maintenance of anaesthesia and undergoing a porto intracaval shunt operation; group C receiving the same drugs as group B but undergoing a simple portal and caval clamping. The following parameters were studied: efficacy of premedication, vital parameters before and after clamping and insertion of the shunt, mean time to clamp and insert the shunt, mean survival time after clamping (group C) or activation of the shunt (groups A and B). Midazolam was significantly better for premedication; there was no statistically significant difference between groups A and B for the vital parameters, for the time necessary to clamp and insert the shunt, for the intraoperative course, and for the mean survival time. The absence of a statistically significant difference between groups could be due to the low number of animals used in the study. There is actually evidence that a correct anaesthesiologic protocol, especially referring to analgesia and fluid management, improves the outcome of operated animals. Surely further studies, possibly conducted on a larger number of animals, are required to evaluate better the results observed and to consider applying these data and this experience to humans. PMID- 11494378 TI - Mandible reconstruction and autogenous frozen bone graft: experimental study on rats. AB - The aim of this study was to evaluate the biological behaviour of a frozen bone graft in orthotopic and heterotopic sites in the rat. The previous experimental study on this subject was published 25 years ago without sufficient detail about the histology and comparison between the orthotopic and ectopic sites. Therefore, being very important for future clinical application, we decided to evaluate the frozen bone graft using rats. The procedure was performed on two groups of five rats each (Charles River). After wide dissection of the inferior border of the mandible from the surrounding muscle, an inferior segmental resection 4 mm in length was performed, taking care not to fracture the superior part and to maintain mucosal integrity. This segment was placed in liquid nitrogen for two periods of 10 minutes each with a third period to allow it to reach room temperature. In the first group (A), the frozen segment was placed ectopically in a gluteal muscle pocket, and in the second group (B), the frozen bone was fixed in the same position in the same mandible. After 1 month of follow-up, the animals were killed, the bone graft was removed, and histology was performed. Results were consistent in both groups. In group A, the segment was surrounded by strong inflammatory reaction, with no vital cells or bone cells, but some vascular penetration. We concluded that there was no bone deposition and no bone rehabitation. In group B, the initial segment was strongly fixed to the remaining mandible, there was an increase of the macroscopic dimension that paralleled the increase in the dimension of the remaining mandible and the growth of the animal. The cortical part had thinned down, the medullary part presented signs of bone deposition as well as bone resorption and vascular penetration. The periosteum from the adjacent normal mandible was growing and covering the frozen bone graft, offering additional stimulus to the bone deposition. In conclusion, the frozen bone graft acts as a normal bone graft. It needs to be placed in contact with vascularised bone and surrounded by well vascularised soft tissue to allow deposition of new bone. If the frozen graft is placed ectopically, it will be surrounded by chronic inflammatory reaction with no bone deposition. PMID- 11494379 TI - Long-term results after lymphatic-venous anastomoses for the treatment of obstructive lymphedema. AB - Over the past 25 years, 665 patients with obstructive lymphedema have been treated with microsurgical lymphatic-venous anastomoses; of these, 446 patients were available for long-term follow-up study. Objective assessment was undertaken by water volumetry and lymphoscintigraphy. Lymphangioscintigraphy, lymphangiography (in patients with gravitational reflux pathology), and echo Doppler were used preoperatively. Subjective improvement was noted in 578 patients (87%). Objectively, volume changes showed a significant improvement in 552 patients (83%), with an average reduction of 67% of the excess volume. Of those patients followed up, 379 patients (85%) have been able to discontinue the use of conservative measures, with an average follow-up of more than 7 years and average reduction in excess volume of 69%. There was a 87% reduction in the incidence of cellulitis after microsurgery. In those patients who improved, drainage resulted in increased softness of the limbs. Peripheral edema (hand and foot) diminished considerably in most patients. These long-term results indicate that lymphatic-venous anastomoses have a place in the treatment of obstructive lymphedema and should be the therapy of choice in patients who are not sufficiently responsive to nonsurgical treatment. Improved results can be expected with earlier operations because patients referred earlier usually have fewer lymphatic alterations. PMID- 11494380 TI - Spleen autotransplantation in mice: a novel experimental model for immunology study. AB - The aim of the experimental model to develop a spleen transplant model in mice to study the role of spleen in autoimmune and transplant rejection. After a midline incision, splenectomy was performed. Four tiny segments were cut from the removed spleen and were rinsed at room temperature in physiological salt solution. The greater omentum was lifted and four omental pockets were created; four thin segments were then placed into the "nests," subsequently marked, and fixed using 8-0 suture. The abdomen was then closed. The duration of the survival time was different among the nine groups (n = 3-3). Tissue samples were taken from the marked areas for histological examination stained with hematoxylin and eosin (H&E). H&E staining demonstrated large, well-circumscribed splenic nests with lymphoid zone and red pulp and well-formed trabecules in the spleen. Among the possible applications of this novel model is the ability to study the role of spleen in autoimmune and organ rejection. PMID- 11494381 TI - The use of vein grafts in the treatment of peripheral lymphedemas: long-term results. AB - This study evaluates long-term results of the treatment of peripheral lymphedemas by the microsurgical reconstructive technique of interposed vein grafts. The technique consists of the use of autologous vein grafts to reconstruct lymphatic pathways where there is a block to the lymphatic circulation of the limb, whether of congenital or acquired etiology. The venous segment represents a sort of "bridge" between afferent and efferent lymphatic collectors (lymphatic-venous lymphatic plasty [LVLA]). The results also proved to have positive long-term effects after microsurgical operation. Follow-up evaluation was performed clinically by water volumetry and instrumentally by lymphangioscintigraphy. With this LVLA technique, peripheral lymphedemas can be treated when derivative lymphovenous shunts cannot be used because of impaired venous circulation in the same lymphedematous limb. The new aspect of the study is that we report long-term clinical and instrumental results. PMID- 11494382 TI - Sequential transplantation induces islet allograft tolerance. AB - Liver transplantation (LT) in some rat strain combinations can induce tolerance to other organ grafts of liver donor strain. The aim of the study was to examine the effect of LT on islet allografts. Islets were isolated by collagenase digestion and purified by dextran-gradient separation. Islets from two donor animals (>2,000) were transplanted under the left kidney capsule of streptozotocin-induced diabetic rats. The liver was implanted orthotopically. The results showed that concurrent LT and islet transplantation (IT) prolonged survival of islet allografts modestly, but islet allografts survived indefinitely when LT was performed before or after IT. LT delayed or reversed ongoing islet graft rejection. IT could trigger rejection in otherwise tolerated liver grafts. We conclude that sequential IT-LT or LT-IT can induce islet allograft tolerance or liver rejection and that the mechanisms involved may differ from those involved with LT simultaneously with either IT or vascularized organ grafts. PMID- 11494383 TI - Confocal imaging of Schwann-cell migration along muscle-vein combined grafts used to bridge nerve defects in the rat. AB - Schwann cells guide axonal regrowth during peripheral nerve repair. In a case of a nerve lesion with substance loss, a graft conduit is necessary to enable axons to reach the distal nerve stump. If a non-nervous autograft is used, the question arises as to the presence and origin of Schwann cells along the grafted tube. We addressed this issue using a tubulization technique based on the use of an autologous vein filled with fresh skeletal muscle for the repair of sciatic nerve defects in the rat. We showed that both ends of the graft were early and progressively colonized by a number of glial fibrillar acid protein immunopositive and S-100 immunonegative cells, an immunocytochemical pattern typical of immature Schwann cells. These cells, which were located in the interstice between grafted skeletal muscle fibers, are mainly organized into long chains oriented along the main axis of the graft and progressively colonize all the graft. Schwann cells coming from the distal nerve end are suitable for being responsible for guiding regeneration of nerve fibers along the graft toward the correct periphery (tissue specificity). PMID- 11494384 TI - Orthotopic liver/small bowel transplantation in rats: a microsurgical model inducing tolerance. AB - Liver cirrhosis in patients with short bowel syndrome is successfully treated in humans by simultaneous liver/small bowel transplantation. However, until now, a clinically relevant experimental rat model for this procedure has not existed. We therefore established a protocol that, for the first time in rats, allows the simultaneous transplantation of arterialized liver and small bowel into an orthotopic position. Short-term immunosuppression induced not only allograft acceptance but tolerance (as demonstrated by indicator heart/skin transplantation). The immunosuppressive dose required to achieve this result was dramatically less than that of protocols for successful small bowel transplantation alone. Immunohistochemistry detected a transient rejection crisis before tolerance. During this crisis, apoptotic recipient-type T lymphocytes, mainly CD8+ cells, accumulated in the liver but not in the small bowel allograft. The initiation of T-cell apoptosis is one possible explanation for the specific immunosuppressive effect of the liver allograft, which also supports the simultaneously transplanted small bowel allograft in our model. PMID- 11494385 TI - Liver microcirculation after selective denervation. AB - Microcirculatory disturbances have been related to a decrease in survival after liver transplant. Because innervation is involved in liver hemodynamics regulation, we decided to evaluate microcirculatory hepatic perfusion. Thirty rats were divided into three groups: denervated (DG), hepatic microsurgical denervation; manipulated (MG), hepatic manipulation; control (CG), laparotomy. Hepatic microcirculation was assessed in the median lobe using laser Doppler flowmetry in the following moments: T(0), after laparotomy and T(1), after denervation; and in the following moments after denervation: T(2), 10 minutes, T(3), 20 minutes, T(4), 30 minutes, T(5), 1 hour, T(6), 1.5 hours, and T(7), 2 hours for DG, and in same moments for MG and CG. DG showed a decrease in hepatic perfusion for 20 minutes after denervation, different from MG and CG. After that, there was recovery in hepatic perfusion in MG and DG (Kruskal-Wallis and Friedman tests). Therefore, denervation and manipulation alter hepatic microcirculation, but denervation promotes a more severe decrease than manipulation. PMID- 11494386 TI - Syngeneic consecutive rat spleen transplantation bearing infantile testis. AB - Seventy-seven Lewis adult rats received autologous testicular implants in the spleen. These spleens were retransplanted in a consecutive transplant fashion as we described earlier. Some were observed as long as 26 to 27 months. Although earlier splenic-testicular transplants showed benign granulosa cell tumors, some of the long-term follow-ups showed a seminoma-like transformation. PMID- 11494387 TI - Gangliosides in rat femoral injury: early effect on intimal hyperplasia. AB - Previous studies demonstrated that some immunosuppressive agents inhibit arterial intimal hyperplasia. Our previous studies demonstrated that gangliosides (Gang) have an immunosuppressive effect on as well as an anti-inflammatory role in the wound-healing process. Therefore, we decided to examine the effect of Gang on intimal hyperplasia. Twenty Wistar isogenic rats received a transverse division of the anterior wall of the femoral artery, followed by suturing using mononylon 10-0 under surgical microscopy and were then divided into two groups: Gang group, 3 mg/kg per day of Gang, and control group, vehicle, intramuscularly from surgery to death (1 and 3 weeks, respectively). Concentric intimal hyperplasia was observed in arteries stained by hematoxylin-eosin in control and Gang groups. However, the media layer did not demonstrate any major alterations. After 3 weeks, the Gang group showed more intimal hyperplasia than the control group. Therefore, because intimal hyperplasia worsened in the presence of Gang after 3 weeks, further studies will be necessary to clarify its role in intimal proliferation. PMID- 11494388 TI - Effects of simultaneous kidney-pancreaticoduodenal transplantation on diabetes induced renal insufficiency in rats. AB - An investigation of the functional and histological changes was done after en bloc kidney-pancreaticoduodenal transplantation (kpdt) in the diabetes-induced, renal insufficient Lewis rats. For donor preparation, an end-to-side portocaval shunt was performed, and the aortic, vena caval segments, and ureter-bladder patch were obtained. They were anastomosed microsurgically to recipient's aorta, vena cava, and bladder in end-to-side fashion. Of 15 diabetes-induced kpdt rats, 14 survived. Two of the 14 surviving rats showed ischemic necrosis. The remaining 12 transplants showed well-preserved glomeruli and Langerhans islets for 5 months postoperatively. Biochemical data comparing diabetic and sham-operated rats (six rats each), six diabetic controls, and 12 kpdt rats showed no significant statistical difference at said observation period. The diabetes-induced kpdt rats showed improvement of following biochemical data: within 1 week postoperatively, the glucose level fell from 300 to 115 mg/dL; BUN level from >20 to <20 mg/dL; the creatinine level from 1.5 to <1.2 mg/dL. The insulin level returned to normal, 1.1 ng/mL, in 2 weeks. The results demonstrate that the kpdt model is an effective and successful operative technique in diabetic rats and may provide effective therapeutic methods for diabetes-induced renal insufficiency. PMID- 11494389 TI - Effectiveness of porto-intracaval shunt to reduce the negative effects of portal and caval clamping in the rabbit. AB - In performing experimental liver surgery, it is difficult to prolong anhepatic time because the animals do not tolerate prolonged portal and caval clamping. To counteract prolonged venous stasis, the authors previously developed a simple porto-intracaval shunt. The shunt consists of a self-constructed inverted Y silicone tube. The effectiveness of this shunt was studied comparing two groups of 10 rabbits with shunt (S) versus those with clamped portal and inferior caval vein (C). In the group of rabbits that underwent porto-intracaval shunt, the results concerning intraoperative mortality, intraoperative increase in distal portal vein pressure, and incidence of the histologic signs of gut damage were clearly improved. The proposed porto-intracaval shunt was therefore effective in reducing some principal negative effects of portal and caval clamping. This type of porto-intracaval shunt can be therefore useful allowing improvement of experimental models concerning liver surgery in little animals. PMID- 11494390 TI - Antigenic load and peripheral chimeric levels in entire and partial liver allograft recipients. AB - Orthotopic partial liver transplantation (PLT) models were developed in rats to explore the unique role of the liver in transplant tolerance. In PVG rats, syngeneic PLT established that surgical reduction to one-third of the liver and orthotopic transplantation permitted survival. Allogeneic PLT in the PVG to DA liver-tolerant model, both 50% and 33%, did not affect the tolerogeneic property of the liver, with all PLT recipients surviving indefinitely. Blood samples taken at various time points for detection of donor cells using flow cytometry showed a steady increase in donor cell chimerism in both PLT and whole liver transplantation (WLT) recipients that persisted throughout the 3-month observation period. At each time point, the level of donor cell chimerism in PLT was higher than that in WLT. We conclude that transplantation of one-third of the liver is compatible with survival in rats. Reduction of antigenic load by means of hepatectomy does not affect the tolerogenic effect of the liver in the PVG to DA LT model because of the remarkable regeneration capability of the liver. Peripheral chimeric levels increase progressively after WLT, suggesting that this is an ongoing immunological phenomenon. The earlier and increased chimerism after PLT may be associated with liver regeneration. PMID- 11494391 TI - Rat auxiliary liver transplantation without portal vein reconstruction: comparison with the portal vein-arterialized model. AB - Auxiliary liver transplantation (ALT) has been reintroduced in clinical cases recently and is now believed to be a viable alternative to orthotopic liver transplantation. To provide a simple rat ALT model for studying the physiological and immunological aspects of the ALT graft, a new ALT was performed, and the comparison between this new model and the portal arterialized one that was reported by other investigators was carried out. At first, we confirmed that liver could tolerate the deprivation of its portal flow well, using a portosystemic shunted rat model. The new rat ALT model, in which the ALT graft obtained its blood inflow only from the hepatic artery, was then performed. Our results demonstrated that 50% of the hepatic artery-alone ALT graft showed almost normal structure histologically at 1 month after grafting, with bile secretion preserved. By contrast, only 8% 1-month graft survival was noted in the portal arterialized group, and all grafts stopped bile secretion 1 week after operation. In conclusion, with arterial blood supply alone, the ALT graft survived and demonstrated normal bile secretion function for more than 1 month. Portal vein arterialization is not an appropriate way to establish the graft's blood supply if no pressure adjustment measures were taken in advance. PMID- 11494392 TI - Nerve regeneration through an epineurial sheath: its functional aspect compared with nerve and vein grafts. AB - Although nerve graft is still the only reliable choice in repair of defects in peripheral nerve structure, it has the disadvantage of donor nerve morbidity and of sometimes being unavailable. It has long been researched in alternate nerve grafts with other materials. Studies have shown that nerves could regenerate across short nerve gaps through various conduits, such as veins, pseudosheaths, and bioabsorbable tubes. Despite encouraging studies, their functional results remain unclear. The present study used 40 rats, in which nerve grafts, vein grafts, and epineurial tubes were placed into 1-cm gaps in sciatic nerves created by resection. In one group, sciatic nerves were denuded of the surrounding epineurium, to assess the possible morbidity caused by epineurial sheath technique. At 2, 4, 8, 12, 20, and 28 weeks, functional assessment of nerve regeneration was performed using walking track analysis. The number of myelinated fibers and fiber diameters was measured and electron microscopic evaluation performed. Functionally, the index values were very close to each other in nerve graft and epineurial sheath groups. Morphometric analysis showed significance between the groups. The result of denuded sciatic nerve group was the same as the base track values. It was concluded that the ready availability of epineurial sheath as a conduit to span short nerve gaps could eliminate the morbidity associated with nerve graft harvest and capitalize on the potential benefits of neurotrophism in directing nerve regeneration. PMID- 11494393 TI - Free tissue transfer in pregnancy: guidelines for perioperative management. AB - A successful free tissue transfer of serratus anterior muscle, to provide coverage for an open ankle defect in a pregnant patient, is described. Microvascular surgery in the presence of a viable pregnancy demands considerations unique to this situation. Although rarely possible, an attempt should be made to plan surgery to coincide with the second trimester, to lessen the risk of anesthesia to the fetus. Maternal positioning, fluid balance, and aspiration precautions need to be critically addressed. Close perioperative monitoring by an obstetrician is essential. The condition of pregnancy results in a hypercoagulable state that may lead to an increased risk of anastomotic failure. The use of anticoagulants results in increased risk of bleeding, not only for the patient but also for the fetus, as well as risk of teratogenic effects. Closely monitored heparin is considered safe in pregnancy as is low molecular-weight dextran and low-dose aspirin. Additional considerations include the use of narcotics and sedatives for comfort postoperatively, as well as antibiotic choices, if indicated. PMID- 11494394 TI - Activation of coagulation and fibrinolysis during reconstructive microsurgery in patients with cancer. AB - Cancer patients are subjected to increased systemic risk of thrombotic events and may therefore be at higher risk of even local thrombosis compromising the outcome of reconstructive microsurgery. Coagulation and fibrinolysis activities were studied serially during and after reconstructive microsurgery in seven patients with oropharyngeal cancer or sarcoma in the extremities. A preoperative hypercoagulable state was found in four patients (elevated fibrinogen, TAT, F1+2, or D-dimer); two of these patients also had a local thrombotic event. In all patients, the plasma markers generally varied perioperatively: fibrinogen decreased, whereas TAT and subsequently D-dimer increased at the end of the operation. However, tPA and PAI-1 remained unaltered, except in the patients with thrombosis in whom PAI-1 activity increased progressively during the operation. F1+2 was also clearly elevated in these two patients at the time of thrombosis. Preoperative assessment of hypercoagulability for this group of patients could be helpful in targeting meticulous antithrombotic protection. PMID- 11494395 TI - Horseradish peroxidase retrograde labeling of primary sensory neurons: a comparison of four intraspinal application methods. AB - The purpose of this study was to optimize the methods of retrograde labeling of sensory neurons in demonstrating the continuity of post-ganglionic primary sensory axons. This was accomplished by comparing four different methods of horseradish peroxidase (HRP) application into the lower thoracic spinal cord of adult rats (level T11). HRP application with a piece of Gelfoam via a dorsal myelotomy (group 1, n = 8), stereotactic injections with a 0.72-mm tip diameter needle (group 2, n = 8), with a 0.24-mm tip needle (group 3, n = 8), and with a 0.08-mm tip glass micropipette (group 4, n = 5). Histological examination of the application site showed that the extent of spinal cord injury was directly proportional to the diameter of the needle tip. The mean number of dorsal root ganglia (DRG) sensory neurons retrogradely stained by HRP differed among the four experimental groups: 77 +/- 45 (SEM) per DRG in group 1, 106 +/- 24 in group 2, 652 +/- 90 in group 3, and 238 +/- 60 in group 4. A significant difference was found between group 3 and the other ones (P < 0.05). Intraspinal injection of HRP with a fine needle (0.24-mm tip diameter) using a stereotactic approach can achieve effective and reliable retrograde labeling of primary sensory neurons. This reproducible method may be useful in studies dealing with regeneration of post-ganglionic primary sensory axons. PMID- 11494396 TI - Rat parathyroid allotransplantation: Influence of MHC antigen expression on graft survival. AB - MHC antigen expression in parathyroid tissue and its influence on graft survival after allogeneic transplantation were investigated using a heterotopic rat transplantation model. MHC class I and II expression in parathyroid tissue of Lewis (LEW), Dark Agouti (DA), and Wistar-Furth (WF) rats was analysed semi quantitatively by using immunohistochemistry. MHC class I expression was strong in DA, moderate in WF, and weak in LEW rats parenchyma, whereas MHC class II expression was negative. In the interstitium of all investigated tissue specimens, the proportion of MHC class II-expressing cells was low. Additionally, four groups were transplanted: 1) LEW to LEW, 2) DA to DA, 3) LEW to DA, and 4) WF to LEW. After syngeneic transplantation, graft survival could be documented over the whole observation period. A median graft survival of 20 (+/-2) days was observed after transplantation from LEW to DA, whereas grafts in the group WF to LEW were rejected after 13 (+/-1) days. The number of intra-graft leucocytes expressing MHC class II molecules was the same in all groups, whereas increased levels of MHC class I in parathyroid tissue before transplantation resulted in a more rapid rejection. These results indicate that immunogenicity of rat parathyroid tissue might be determined by the amount of MHC class I expressed in donor parenchymal cells. Further experiments are necessary to validate this observation. PMID- 11494397 TI - Use of colour duplex Doppler imaging in the postoperative assessment of buried free flaps. AB - The postoperative assessment of free flaps is essential to identify and act on signs of incipient flap failure. Where the flap is completely buried, this becomes almost impossible unless part of the flap is exteriorised or an overlying skin window is used. Alternatively, complicated and often impractical monitoring devices have been advocated, but these have failed to gain widespread acceptance. A simpler solution to this problem has been evaluated in a series of patients using colour duplex Doppler imaging. This re-appraisal of a previously reported technique has been facilitated by updated technology in diagnostic radiology. Duplex Doppler imaging was confirmed as an accurate, non-invasive, and inexpensive tool for the postoperative measurement of blood flow within the pedicles of five buried free flaps in four patients undergoing surgery in our unit. PMID- 11494398 TI - Sex differences in [123I]beta-CIT SPECT measures of dopamine and serotonin transporter availability in healthy smokers and nonsmokers. AB - Nicotine and other constituents of tobacco smoke elevate dopamine (DA) and serotonin (5-HT) levels in brain and may cause homeostatic adaptations in DA and 5-HT transporters. Since sex steroids alter DA and 5-HT transporter expression, the effects of smoking on DA and 5-HT transporter availability may differ between sexes. In the present study, DA and 5-HT transporter availabilities were quantitated using single photon emission computed tomography (SPECT) imaging approximately 22 h after bolus administration of [123I]beta-CIT, an analog of cocaine which labels DA and 5-HT transporters. Forty-two subjects including 21 pairs of age-, race-, and gender-matched healthy smokers and nonsmokers (12 female and 9 male pairs) were imaged. Regional uptake was assessed by the outcome measures, V3", which is the ratio of specific (i.e., ROI-cerebellar activity) to nondisplaceable (cerebellar) activity, and V3, the ratio of specific to free plasma parent. Overall, striatal and diencephalic [123I]beta-CIT uptake was not altered by smoking, whereas brainstem [123I]beta-CIT uptake was modestly higher (10%) in smokers vs. nonsmokers. When subgrouped by sex, regardless of smoking status, [123I]beta-CIT uptake was higher in the striatum (10%), diencephalon (15%), and brainstem (15%) in females vs. males. The sex*smoking interaction was not significant in the striatum, diencephalon, or brainstem, despite the observation of 20% higher brainstem [123I]beta-CIT uptake in male smokers vs. nonsmokers and less than a 5% difference between female smokers and nonsmokers. The results demonstrate higher DA and 5-HT transporter availability in females vs. males and no overall effect of smoking with the exception of a modest elevation in brainstem 5-HT transporters in male smokers. Although these findings are preliminary and need validation with a more selective 5-HT transporter radiotracer, the results suggest that brainstem 5-HT transporters may be regulated by smoking in a sex-specific manner. PMID- 11494399 TI - Age-related changes of dopamine D1-like and D2-like receptor binding in the F344/N rat striatum revealed by positron emission tomography and in vitro receptor autoradiography. AB - To clarify age-related changes in dopamine D1-like and D2-like receptor binding in the striatum, positron emission tomography (PET) and in vitro receptor autoradiography (in vitro ARG) were performed using F344/N rats of various ages (6, 12, 18, and 24 months). In the PET study, [11C]SCH23390 and [11C]raclopride were used to image dopamine D1-like receptors and dopamine D2-like receptors, respectively, while [3H]SCH23390 and [3H]raclopride were used for the in vitro ARG study. With PET, we calculated the binding potential (= k3/k4, Bmax/Kd) of [11C]SCH23390 and [11C]raclopride in the striatum according to the curve fitting (CF) and the Logan plot (LP) methods. The binding potential of [11C]SCH23390 in the striatum demonstrated significant decrease as a function of age (max. decrease -26%) by the LP method, while this was not observed in the data analyzed by the CF method. In contrast, the binding potential of [11C]raclopride in the striatum decreased significantly with age by both the CF (max. decrease -28%) and the LP (max. decrease -36%) methods. However, no significant difference by either method was observed in rats between 6 and 12 months old using either ligand. In the in vitro ARG study, the specific binding (fmol/mg tissue) of [3H]SCH23390 and [3H]raclopride in the striatum were determined. Both [3H]SCH23390 and [3H]raclopride binding declined considerably with age as noted by comparing 12, 18, and 24-month-old rats against those 6 months old (max. decrease -29% and 31%, respectively). The substantial difference in binding shown in 12-month-olds in comparison with 6-month-olds using either ligand with in vitro ARG was in contrast with the PET results. These distinctions between the PET and the in vitro ARG studies may be attributed to the receptor microenvironment created under these experimental conditions. The results indicate that PET with LP analysis is useful in obtaining age-related changes of D1-like and D2-like receptor binding in the striatum of living rats. PMID- 11494400 TI - Biphasic modulation of NMDA-induced responses in pyramidal cells of the medial prefrontal cortex by Y-931, a potential atypical antipsychotic drug. AB - Similar to the effects produced by the atypical antipsychotic drugs (APDs) clozapine and olanzapine, Y-931 [8-fluoro-12-(4-methylpiperazin-1-yl)-6H [1]benzothieno[2,3-b][1,5]benzodiazepine maleate, a purported atypical APD] effectively facilitated N-methyl-D-aspartate (NMDA)-induced, but not (+/-)-alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-evoked, responses in pyramidal cells of the rat medial prefrontal cortex (mPFC). Similar to olanzapine and clozapine, the concentration-response curve of Y-931 in these experiments was biphasic. At present, the mechanisms behind the biphasic modulatory actions of Y 931 and olanzapine on NMDA-induced currents in the mPFC are not clear. In addition to augmenting NMDA responses, Y-931 prevented the phencyclidine (PCP) induced block of the NMDA responses and increased the amplitudes and durations of excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation of the forceps minor. Overall, our findings suggest that APDs, particularly the atypical ones, share a common property in that they facilitate NMDA receptor-mediated transmission in the mPFC and perhaps other functionally related limbic structures as well, which could be the cellular basis for their ability to alleviate some schizophrenic negative symptoms and cognitive dysfunctions. PMID- 11494401 TI - Differential time-course profiles of dopamine release and uptake changes induced by three dopamine uptake inhibitors. AB - Using real-time voltammetry, we compared the effects of cocaine (1.0, 3.0, or 10 microM), WIN 35428 (0.1, 0.5, or 2.0 microM), and nomifensine (0.2, 1.0, or 5.0 microM) on electrically evoked dopamine release and uptake in the rat accumbens slice. The time course for onset and offset of the drug effects were determined by perfusing single drug concentration for 30 min, followed by a 60-min washout. Cocaine elicited a rapid, concentration-independent increase in dopamine release and a more gradual, concentration-dependent inhibition of uptake. During washout, uptake inhibition rapidly abated to near baseline values. During the same period, the potentiation of dopamine release exhibited a slower offset for all concentrations and, for 10 microM cocaine, was even greater than that observed during drug perfusion ("rebound" increase). The release rebound was not observed during continuous 90-min perfusion, verifying that cocaine washout per se was a sufficient condition. Selective D1 or D2 antagonists (0.5 microM SCH 39166 or 2 microM sulpiride, respectively) were without effect on cocaine-induced release alterations. WIN 35428 and nomifensine induced similar changes in dopamine kinetics during perfusion. However, in contrast to cocaine, no consistent release rebound was observed during their washout. For 2 microM WIN 38425, washout and continuous perfusion groups exhibited similar changes in dopamine release and uptake. The time-course mismatch between uptake inhibition and DA release potentiation as well as release rebound during washout suggests that altered dopamine release might play a role in behavioral effects of cocaine. PMID- 11494402 TI - Plasmalemmal mu-opioid receptor distribution mainly in nondopaminergic neurons in the rat ventral tegmental area. AB - Opiate-evoked reward and motivated behaviors reflect, in part, the enhanced release of dopamine produced by activation of the mu-opioid receptor (muOR) in the ventral tegmental area (VTA). We examined the functional sites for muOR activation and potential interactions with dopaminergic neurons within the rat VTA by using electron microscopy for the immunocytochemical localization of antipeptide antisera raised against muOR and tyrosine hydroxylase (TH), the synthesizing enzyme for catecholamines. The cellular and subcellular distribution of muOR was remarkably similar in the two major VTA subdivisions, the paranigral (VTApn) and parabrachial (VTApb) nuclei. In each region, somatodendritic profiles comprised over 50% of the labeled structures. MuOR immunolabeling was often seen at extrasynaptic/perisynaptic sites on dendritic plasma membranes, and 10% of these dendrites contained TH. MuOR-immunoreactivity was also localized to plasma membranes of axon terminals and small unmyelinated axons, none of which contained TH. The muOR-immunoreactive axon terminals formed either symmetric or asymmetric synapses that are typically associated with inhibitory and excitatory amino acid transmitters. Their targets included unlabeled (30%), muOR-labeled (25%), and TH labeled (45%) dendrites. Our results suggest that muOR agonists in the VTA affect dopaminergic transmission mainly indirectly through changes in the postsynaptic responsivity and/or presynaptic release from neurons containing other neurotransmitters. They also indicate, however, that muOR agonists directly affect a small population of dopaminergic neurons expressing muOR on their dendrites in VTA and/or terminals in target regions. PMID- 11494403 TI - Vesicular neurotransmitter transporters in Huntington's disease: initial observations and comparison with traditional synaptic markers. AB - Markers of identified neuronal populations have previously suggested selective degeneration of projection neurons in Huntington's disease (HD) striatum. Interpretations are, however, limited by effects of compensatory regulation and atrophy. Studies of the vesicular monoamine transporter type-2 (VMAT2) and of the vesicular acetylcholine transporter (VAChT) in experimental animals indicate that they are robust markers of presynaptic integrity and are not subject to regulation. We measured dopamine and acetylcholine vesicular transporters to characterize the selectivity of degeneration in HD striatum. Brains were obtained at autopsy from four HD patients and five controls. Autoradiography was used to quantify radioligand binding to VMAT2, VAChT, the dopamine plasmalemmal transporter (DAT), benzodiazepine (BZ) binding sites, and D2-type dopamine receptors. The activity of choline acetyltransferase (ChAT) was determined as an additional marker of cholinergic neurons. Autoradiograms were analyzed by video assisted densitometry and assessment of atrophy was made from regional structural areas in the coronal projection. Striatal VMAT2, DAT, and VAChT concentrations were unchanged or increased, while D2 and BZ binding and ChAT activity were decreased in HD. After atrophy correction, all striatal binding sites were decreased. However, the decrease in ChAT activity was 3-fold greater than that of VAChT binding. In addition to degeneration of striatal projection neurons, there are losses of extrinsic nigrostriatal projections and of striatal cholinergic interneurons in HD on the basis of vesicular transporter measures. There is also markedly reduced expression of ChAT by surviving cholinergic striatal interneurons. PMID- 11494404 TI - Prenatal exposure to cocaine reduces the number and enhances reactivity of A10 dopaminergic neurons to environmental stress. AB - Prenatal exposure to cocaine has been shown to result in poor cognitive performance in the resulting offspring in humans and laboratory animals. The underlying biochemical changes that contribute to these behavioral effects are not known but have been proposed to involve changes in dopaminergic function. In these studies, we exposed rats to cocaine in utero using the clinically relevant intravenous model and report a mean loss of 24.8% of the tyrosine hydroxylase immunoreactive, presumed dopaminergic, neurons in the A10, but not A9 and A8, cell groups of the young adult offspring. Additionally, in prenatal cocaine exposed rats dopaminergic neurons in the ventral, midline A10, and lateral A9 regions demonstrated a hyperreactivity to environmental stress, as measured by activation of the immediate-early gene, Fos. Mild, intermittent footshock did not further increase the number of dopamine neurons expressing Fos in prenatal cocaine-exposed rats, as it did in the prenatal saline controls. Because the exposure to cocaine took place during development, other potential changes in dopaminergic and nondopaminergic neuronal systems could result from the cocaine induced reduction in numbers of A10 dopamine neurons. We hypothesize that a perinatal loss of A10 dopamine neurons, and subsequent developmental changes, contributes to a dysregulation of the adult mesoprefrontal system, resulting in the reported cognitive deficits. PMID- 11494405 TI - Maturational increases in c-fos expression in the ascending dopamine systems. AB - The unique maturational period of adolescence is replete with numerous changes in anatomy and function that may yield clues as to why drug abuse emerges at this stage. The behavioral effects of amphetamine are diminished during periadolescence (35 days) relative to younger (21 days) and older (>60 days) rats, prompting us to examine amphetamine effects on neuronal activation with the immediate early gene, c-fos. Amphetamine (1 and 5 mg/kg, i.p.) increased c-fos immunoreactivity in rats 21, 35, and 60 days of age in a dose-dependent manner. When expressed as a percentage of vehicle for each age, amphetamine-induced effects on c-fos immunoreactivity were higher at 21 days of age compared with the effects at 35 and 60 days of age in the nucleus accumbens core and shell, striatum, and prefrontal cortex. These data provide a possible reason as to why stimulants produce dysphoria in children, before transitioning to euphoria during adolescence. Implications of these results are discussed for stimulant use in a pediatric population and the development of drug abuse. PMID- 11494406 TI - Modulation of voltage-dependent calcium currents by serotonin in acutely isolated rat amygdala neurons. AB - The modulation of voltage-dependent calcium currents (I(Ca)) by serotonin (5-HT) was studied in rat acutely dissociated amygdala neurons using whole-cell patch clamp recording techniques. 5-HT inhibited I(Ca) in a concentration-dependent manner with a ED50 of approximately 1 microM and a maximal inhibition of approximately 50%. The inhibition was mimicked by the selective 5-HT1A agonist 8 hydroxy-dipropylaminotetralin (8-OH-DPAT) and was reduced by the 5-HT1A antagonist NAN-190, indicating its mediation by 5-HT1A receptors. Pretreatment of neurons with the alkylating agent N-ethylmaleimide (NEM) or pertussis toxin (PTX) markedly reduced the action of 5-HT. The modulation was partially reversed by strong depolarization and was not seen in cell-attached patches when the agonist was applied outside the recorded patch, suggesting a membrane-delimited, G protein-mediated signaling pathway. Nimodipine (1 microM) reduced the I(Ca) by approximately 30% without reducing inhibition of current by 5-HT significantly, ruling out L-type channels as the target of modulation. 5-HT-mediated inhibition after exposure to omega-conotoxin-GVIA (omega-CgTX, 1 microM) or omega-agatoxin IV (omega-AgTX, 200 nM), which blocked 26% and 21% of the total I(Ca), respectively, was significantly decreased, suggesting involvement of the N- and P/Q-type channels. In the combined presence of omega-CgTX and omega-AgTX, 5-HT still caused a small but significant reduction of I(Ca), suggesting a possible involvement of R-type channels. Stimulation of beta-adrenergic receptor with isoproterenol (Iso) or activation of adenylyl cyclase with forskolin resulted in an enhancement of I(Ca). 5-HT caused the same degree of inhibition with or without Iso or forskolin pretreatment. On the other hand, application of 8-OH DPAT inhibited I(Ca) and blocked Iso- and Sp-cAMPS-induced enhancement. These results provide the first evidence showing a dominant effect of 5-HT-mediated inhibition over Iso-mediated enhancement of I(Ca). PMID- 11494407 TI - Dopamine denervation induces neurotensin immunoreactivity in GABA-parvalbumin striatal neurons. PMID- 11494408 TI - Early expression of glutamate transporter proteins in ramified microglia after controlled cortical impact injury in the rat. AB - Traumatic brain injury is followed by increased extracellular glutamate concentration. Uptake of glutamate is mainly mediated by the glial glutamate transporters GLAST and GLT-1. Extent and distribution of GLAST and GLT-1 were studied in a rat model of controlled cortical impact injury (CCII). Western Blot analysis revealed lowest levels of GLAST and GLT-1 with a decrease by 40%-54% and 42%-49% between 24 and 72 h posttrauma. By 8 h after CCII, CSF glutamate levels were increased (10.5 microM vs. 2.56 microM in controls; P < 0.001), reaching maximum values by 48 h. A significant increase in de novo GLAST and GLT-1 expressing ramified microglia was observed within 4 h, reached a stable level by 48 h, and remained high up to 72 h after CCII. Furthermore, ramified microglia de novo expressed the neuronal glutamate transporter EAAC1 after CCII. Following CCII, GLAST/GLT-1 and GFAP coexpressing astrocytes were immediately reduced, reaching minimum levels within 8 h. This reduction of expression could be either due to protein downregulation or loss of astrocytes. At 72 h, a marked population of GLAST- and GLT-1-positive reactive astrocytes appeared. These results support the hypothesis that reduced astrocytic GLAST and GLT-1 protein levels following CCII contribute to evolving secondary injury. Microglia are capable of de novo expressing glutamate transporter proteins, indicating that the expression of glial and neuronal glutamate transporters is not restricted to a specific glial or neuronal lineage. Ramified microglia may play an important compensatory role in the early regulation of extracellular glutamate after CCII. PMID- 11494409 TI - Amphotericin B potentiates the activation of inducible nitric oxide synthase and causes nitric oxide-dependent mitochondrial dysfunction in cytokine-treated rodent astrocytes. AB - Because the neurotoxic effects of the antifungal drug amphotericin B (AMB) closely resemble those ascribed to the highly reactive gaseous free radical nitric oxide (NO), we investigated the effect of AMB on NO production in rodent astrocytes. AMB caused a dose-dependent increase of NO generation in interferon gamma (IFN-gamma)-stimulated rat and mouse astrocytes, as well as in IFN-gamma + tumor necrosis factor-alpha (TNF-alpha)-activated rat astrocytoma cell line C6. Treatment of rat astrocytes with AMB markedly potentiated IFN-gamma-triggered expression of mRNA for iNOS, but not for its transcription factor IRF-1. The activation of transcription factor NF-kappaB was apparently required for AMB induced iNOS mRNA expression, as the latter was abolished by NF-kappaB inhibitors: pyrrolidine dithiocarbamate and MG132. AMB-mediated enhancement of astrocyte NO production was partly dependent on endogenous IL-1, as shown by partial inhibition of AMB effect with IL-1 receptor antagonist. IFN-gamma + AMB treatment led to reduction of astrocyte mitochondrial respiration (measured by MTT assay) that has been completely reverted by selective iNOS inhibitor aminoguanidine. AMB toxicity toward IFN-gamma-stimulated astrocytes was dependent on both AMB and NO action, since AMB and NO-releasing substance SNP synergized in inducing astrocyte mitochondrial dysfunction. These results suggest that the enhancement of cytokine-induced iNOS activation in astrocytes and the subsequent release of high amounts of NO might be at least partly responsible for AMB neurotoxicity. PMID- 11494410 TI - Effect of elevated K(+), hypotonic stress, and cortical spreading depression on astrocyte swelling in GFAP-deficient mice. AB - Glial fibrillary acidic protein (GFAP) is the main component of intermediate filaments in astrocytes. To assess its function in astrocyte swelling, we compared astrocyte membrane properties and swelling in spinal cord slices of 8- to 10-day-old wild-type control (GFAP(+/+)) and GFAP-knockout (GFAP(-/-)) mice. Membrane currents and K(+) accumulation around astrocytes after a depolarizing pulse were studied using the whole-cell patch-clamp technique. In vivo cell swelling was studied in the cortex during spreading depression (SD) in 3 to 6 month-old animals. Swelling-induced changes of the extracellular space (ECS) diffusion parameters, i.e., volume fraction alpha and tortuosity lambda, were studied by the real-time iontophoretic tetramethylammonium (TMA(+)) method using TMA(+)-selective microelectrodes. Morphological analysis using confocal microscopy and quantification of xy intensity profiles in a confocal plane revealed a lower density of processes in GFAP(-/-) astrocytes than in GFAP(+/+) astrocytes. K(+) accumulation evoked by membrane depolarization was lower in the vicinity of GFAP(-/-) astrocytes than GFAP(+/+) astrocytes, suggesting the presence of a larger ECS around GFAP(-/-) astrocytes. Astrocyte swelling evoked by application of 50 mM K(+) or by hypotonic solution (HS) produced a larger increase in [K(+)](e) around GFAP(+/+) astrocytes than around GFAP(-/-) astrocytes. No differences in alpha and lambda in the spinal cord or cortex of GFAP(+/+) and GFAP(-/-) mice were found; however, the application of either 50 mM K(+) or HS in spinal cord, or SD in cortex, evoked a large decrease in alpha and an increase in lambda in GFAP(+/+) mice only. Slower swelling in GFAP(-/-) astrocytes indicates that GFAP and intermediate filaments play an important role in cell swelling during pathological states. PMID- 11494411 TI - Macrophage depletion impairs oligodendrocyte remyelination following lysolecithin induced demyelination. AB - An association between macrophages and remyelination efficiency has been observed in a variety of different models of CNS demyelination. In order to test whether this association is causal or coincidental, we have examined the effects of macrophage depletion on the rate of remyelination of lysolecithin-induced demyelination in the spinal cord of young adult female rats. Macrophage depletion was achieved by reducing the monocyte contribution to the macrophages within the lesion using the clodronate-liposome technique. This technique not only resulted in a decrease in Ox-42-positive cells in the spleen of treated animals but also in the levels of macrophage scavenger receptor type B mRNA expression within the demyelinating lesion. In animals treated with clodronate-liposomes throughout the remyelination process, there was a significant decrease in the extent of oligodendrocyte remyelination at 3 weeks after lesion induction, but no effect on Schwann cell remyelination. If macrophage depletion was delayed until the second half of the remyelination phase, then there was no effect on the repair outcome, implying that macrophages are required for the early stages of CNS remyelination. The results of this study indicate that the macrophage response is an important component of successful CNS remyelination and that approaches to the treatment of demyelinating disease based on inhibition of the inflammatory response may also impair regenerative events that follow demyelination. PMID- 11494412 TI - Electrophysiological alterations and upregulation of ATP receptors in retinal glial Muller cells from rats infected with the Borna disease virus. AB - Infection with the neurotropic Borna disease virus (BDV) causes an immune mediated neurological disease in a broad range of species. In addition to encephalitis, BDV-infected Lewis rats develop a retinitis histologically characterized by the loss of most retinal neurons. By contrast, the dominating retinal macroglia, the Muller cells, do not degenerate. It is known from several models of neurodegeneration that glial cells may survive but undergo significant alterations of their physiological parameters. This prompted us to study the electrophysiology and ATP-induced changes of intracellular Ca(2+)-concentration ([Ca(2+)](i)) in Muller cells from BDV-infected rat retinae. Freshly isolated cells were used for whole-cell patch-clamp recordings. Whereas neither zero current potentials nor membrane resistances showed significant alterations, the membrane capacitance increased in cells from BDV-infected rats during survival times of up to 8 months. This process was accompanied by a decrease in K(+) current densities. Muller cells from BDV-infected rats were characterized by expression of a prominent fast-inactivating A-type K(+) current which was rarely found in control cells. Moreover, the number of cells displaying Na(+) currents was slightly increased after BDV-infection. ATP evoked increases in [Ca(2+)](i) in Muller cells within retinal wholemounts of both control and BDV-infected animals. However, the number of ATP-responding isolated cells increased from 24% (age-matched controls) to 78% (cells from animals > or =18 weeks after infection). We conclude that in BDV-induced retinopathy, reactive rat Muller cells change their physiological parameters but these changes are different from those in Muller cells during proliferative vitreoretinopathy in man and rabbit. PMID- 11494413 TI - Human MBP-specific T cells regulate IL-6 gene expression in astrocytes through cell-cell contacts and soluble factors. AB - One of the distinctive features of multiple sclerosis (MS) attacks is homing to the CNS of activated T cells able to orchestrate humoral and cell-based events, resulting in immune-mediated injury to myelin and oligodendrocytes. Of the complex interplay occurring between T cells and CNS constituents, we have examined some aspects of T-cell interactions with astrocytes, the major components of the glial cells. Specifically, we focused on the ability of T cells to regulate the gene expression of interleukin-6 (IL-6) in astrocytes, based on previous evidence showing the involvement of this cytokine in CNS disorders. We found that T-cell adhesion and T-cell soluble factors induce IL-6 gene expression in U251 astrocytes through distinct signaling pathways, respectively, resulting in the activation of NF-kappaB and IRF-1 transcription factors. In a search for effector molecules at the astrocyte surface, we found that alpha3beta1 integrins play a role in NF-kappaB activation induced by T-cell contact, whereas interferon gamma (IFN-gamma) receptors dominate in IRF-1 induction brought about by T-cell derived soluble factors. Similar phenomena were observed also in normal fetal astrocyte cultures. We therefore propose that through astrocyte induction, T cells may indirectly regulate the availability of a cytokine which is crucial in modulating fate and behavior of cell populations involved in the pathogenesis of MS inflammatory lesions. PMID- 11494414 TI - Continuous interleukin-6 application in vivo via macroencapsulation of interleukin-6-expressing COS-7 cells induces massive gliosis. AB - The inflammatory cytokine interleukin-6 (IL-6) was found in senile plaques of Alzheimer's patients and might be involved in the pathology of Parkinson's disease and multiple sclerosis. Interestingly, an astocytosis is also found in these neurodegenerative disorders. To evaluate the direct effects of IL-6 in vivo on glial cells, we created a new in vivo model. IL-6 and mock-transfected (control group) COS-7 cells were encapsulated in a poly-acryl-nitril membrane for implantation into the rat striatum. Afterward, the host immune reaction to the membrane without encapsulated cells and the biological action of IL-6-producing capsules was evaluated. Animals with an implanted membrane without cells showed a moderate astrocytosis 5 days after the operation. Furthermore, microglia and T cells could be detected and after 30 days the astrocytosis decreased to a small layer around the membrane. In comparison to the control group, which received a sham operation, our results demonstrate that the response of glial cells is caused by the mechanical damage of the surgical procedure itself rather than due to the introduced membrane material. In contrast, we found a massive proliferation and activation of astrocytes and microglia after 10 days by IL-6 secreting capsules, indicating that IL-6 is involved in the induction of gliosis. Control animals that received encapsulated mock-transfected COS-7 cells showed only a weak response. These data point to an involvement of IL-6 in the proliferation and activation of glial cells as seen in neurodegenerative disorders. PMID- 11494416 TI - Anaesthesia and hair fashion. PMID- 11494417 TI - An unexpected and preventable cause of failed epidural analgesia. PMID- 11494415 TI - Multiple compartments with different metabolic characteristics are involved in biosynthesis of intracellular and released glutamine and citrate in astrocytes. AB - The metabolism of glucose and lactate was investigated in cultured mouse cerebellar astrocytes, a culture preparation consisting of a homogeneous population of cells, by incubating the cells in a medium containing either [U (13)C]glucose or [U-(13)C]lactate in combination with unlabeled lactate and glucose, respectively. After the incubation, cell extracts and media were analyzed by GC/MS (gas chromatography/mass spectrometry) for labeling patterns in aspartate, glutamate, and glutamine, as well as the tricarboxylic acid (TCA) cycle constituents citrate and fumarate. Triple labeling of extracellular citrate exceeded that of double labeling from [U-(13)C]glucose. This was not the case when lactate was the labeled precursor. These results indicate that pyruvate carboxylation in biosynthesis of releasable citrate was less prominent from lactate compared with glucose. As observed in the case of extracellular citrate, triple labeling of intracellular aspartate was higher than double labeling when [U-(13)C]glucose was the precursor, but not with [U-(13)C]lactate as precursor. The pattern of labeling in citrate was different intra- and extracellularly and the extent of labeling extracellularly was 10 times higher using [U-(13)C]glucose compared with [U-(13)C]lactate. However, the intracellular citrate labeling from [U-(13)C]glucose only exceeded that originating from labeled lactate by a factor of two. This is in contrast to the labeling pattern obtained for glutamine, since intracellularly this was equally prominent using [U-(13)C]glucose and [U (13)C]lactate as substrates. Moreover, extracellularly the labeling was only slightly higher when using [U-(13)C]glucose compared with [U-(13)C]lactate. Intracellular glutamate and extracellular glutamine exhibited similar incorporation patterns with regard to double compared with triple labeling and the extent of incorporation of label from [U-(13)C]lactate compared with [U (13)C]glucose. It should be noted that the main intracellular pools of citrate and glutamine were compartmentalized; i.e., releasable citrate and glutamine exhibited a labeling pattern distinctly different from that of their intracellular pools. Moreover, carboxylation of pyruvate using glucose as the precursor was more important for biosynthesis of releasable glutamine and citrate, compared with their intracellular pools. Based on the results a model of multiple compartments is suggested. The compartments differ with regard to utilization of lactate and glucose, synthetic pathways for TCA cycle intermediates and amino acids, particularly citrate and glutamine, as well as the contents of different metabolites. PMID- 11494418 TI - Epidural haematoma and timing of heparin prophylaxis. PMID- 11494419 TI - Damage to the conus medullaris following spinal anaesthesia: 1. PMID- 11494420 TI - Damage to the conus medullaris following spinal anaesthesia: 2. PMID- 11494421 TI - Damage to the conus medullaris following spinal anaesthesia: 3. PMID- 11494423 TI - Pressure sores: blame the mobile epidural! PMID- 11494424 TI - Combined spinal-epidural as an alternative method of anaesthesia for a sigmoid colectomy. PMID- 11494425 TI - Predicting caudal epidural analgesia using nerve stimulation. PMID- 11494426 TI - Vertical infraclavicular block and myasthenia gravis. PMID- 11494427 TI - Use of the laryngeal mask is not contraindicated for laparoscopic cholecystectomy. PMID- 11494429 TI - Adrenaline 0.5%? PMID- 11494432 TI - Shocking postoperative visits. PMID- 11494431 TI - Safety, cost and predrawn emergency drugs. PMID- 11494434 TI - Malignant hyperpyrexia and the laryngeal mask airway. PMID- 11494435 TI - Why KY? PMID- 11494436 TI - Another problem with reusable tubes. PMID- 11494438 TI - Reliability of Mallampati classification in upper airway pathology. PMID- 11494439 TI - [Demographic aspects of prehospital diagnosis of myocardial infarction]. AB - AIM: To assess results of prehospital diagnosis of myocardial infarction (MI) in patients of different age and gender. MATERIALS AND METHODS: The results are available of prehospital diagnosis of MI in 1500 patients in 1992-1996. 49.5 and 32.3% patients (females and males, respectively) were over 75 years of age. In this group of patients MI was atypical and repeated in 34.4 and 60.6% of cases. Atypicity was more frequent in females. RESULTS: No statistically significant differences were found in frequency of diagnostic errors in different demographic groups of patients. For 5 years there was a trend to hyperdiagnosis of MI, especially in young patients and males. Late diagnosis occurred more rarely in both males and females. CONCLUSION: Clinical features of MI in elderly patients and females are essential for the number of diagnostic errors especially in high medical qualification. PMID- 11494440 TI - [State of the upper respiratory tract in patients with stage I arterial hypertension]. AB - AIM: To study morphofunctional features of the upper airways (UAW) in wakeful patients (juveniles with arterial hypertension stage I) and genetic predisposition to UAW obstruction in sleeping patients. MATERIAL AND METHODS: The examination of 18 juveniles with arterial hypertension stage I and II normotensive juveniles included measurements of body mass index (BMI), circumference of the neck (CN), conduction of lateral cephalometry, MR imaging of UAW tissues, evaluation of nasal breathing, smoking habits and collection of family disease history. All the patients were examined by otorhinolaryngologist. RESULTS: Hypertensive juveniles appeared to have significantly higher values of BMI, CN, ANB, NAPg angles, distance between the hyoid bone and the mandible, anteroposterior tongue length in lateral projection, less values of SNPg angle (p < 0.05). Relatives of hypertensive juveniles had significantly greater prevalence of hypertension, snoring and their combination. CONCLUSION: In arterial hypertension there are some anatomofunctional grounds to UAW resistance in sleep. In addition to hereditary predisposition to hypertension, this may underlie development of wakeful systemic hypertension. PMID- 11494441 TI - [Treatment and prevention of cognitive dysfunction in patients with arterial hypertension and atherosclerosis: results of a randomized double-blind placebo controlled trial of cerebrolysin]. AB - AIM: To assess therapeutic and prophylactic effect of large-dose cerebrolysin (15 ml/day for 28 days) in hypertensive and atherosclerotic patients with cognitive disorders. MATERIAL AND METHODS: Cerebrolysin was given annually (15 ml/day for 28 days) for 2 years to 42 patients in a randomized double-blind placebo controlled study. The effect was stated by clinical status, neuropsychological and neurophysiological data. RESULTS: In mild disturbances of cognitive functions in patients with arterial hypertension and atherosclerosis courses of cerebrolysin with one-year interval produce stable improvement of subjective status, productivity of memory, attention and thinking which persist for at least a year after the course. The clinical data agree with positive trend in neurophysiological parameters of cognitive component of the response of evoked potentials P-300. CONCLUSION: A course of 28-day annual treatment with cerebrolysin (15 ml/day) of patients with mild defects of cognitive functions stabilizes the process, leads to regression of cognitive disorders predicting vascular dementia. PMID- 11494442 TI - [Free radical scavenging activity of sulfonylureas: a clinical assessment of the effectiveness of gliclazide]. AB - In long-term clinical studies the beneficial effects of gliclazide on platelets have been related to a reduction in oxidative stress. This property is because of gliclazide's free radical scavenging ability that relates to the unique amino azabicyclo-octane ring, which is grafted on to the sulfonylurea. During a blinded clinical trial, the possible effects of gliclazide were assessed in 30 non insulin-dependent diabetic patients. All patients had been treated for diabetes for more than 2 years (mean 8 years) and had been established on glibenclamide for over 2 years with or without adjunctive metformin therapy. Patients were studied for 6 months and randomized to continue either their present dose of glibenclamide or to be converted to an equipotent dose of gliclazide. Measurements were taken of hemostatic variables, the oxidative status of the plasma, and the redox status, both extracellularly as plasma albuminthiols (PSH) and lipid peroxides, and intracellularly as red blood cell superoxide dismutase activity (SOD). At 3 months, diabetic control was unaltered, but there were significant improvements in the oxidative status of the gliclazide-treated patients. Lipid peroxides decreased (8.3 +/- 1.1 to 7.0 +/- 0.06 mumol/l, P < 0.01) and red blood cell SOD increased (135 +/- 21 to 152 +/- 36 micrograms/ml, P < 0.05). PSH levels were unaltered at 453 +/- 38 mumol/l, whereas they had decreased significantly in the glibenclamide patients (414 +/- 34 mumol/l, P < 0.05), resulting in a significant difference between the 2 treatment groups (P < 0.004). Platelet reactivity to collagen also improved in the gliclazide-treated patients, decreasing from 65.1% +/- 14% to 50.8 +/- 24% (p < 0.01). The reactivity of the platelets remained unaltered in the glibenclamide patients. At 6 months, the significant differences between the 2 treatment groups remained. Hence, gliclazide was shown in a clinical study to have free radical scavenging activity independent of glycemic control. PMID- 11494443 TI - [Experience in the use of vessel due F (sulodexide) in patients with suppurative necrotic forms of diabetic foot without critical ischemia]. AB - AIM: To evaluate effectiveness of sulodexide in patients with pyonecrotic complications of diabetic foot. MATERIAL AND METHOD: Sulodexide was given to 15 patients which were examined for blood fibrinogen, foot tissues saturation with oxygen, microbic contamination of the wound tissue. In addition, ultrasound dopplerography of foot arteries, laser doppler flowmetry were performed. RESULTS: Fibrinogen in peripheral blood fell, arteriovenous shunting diminished, capillary blood flow and oxygen saturation of the tissues improved. CONCLUSION: Even early sulodexide treatment is effective at different stages of the pathological process in diabetic patients with severe pyonecrotic lesion of the lower limbs as it recovers microcirculatory blood flow. PMID- 11494444 TI - [Aspects of diastolic function of the heart in vibration disease]. AB - AIM: To study effects of occupational vibrations on the myocardium, to elucidate early alterations in cardiac performance and to introduce approaches to their correction. MATERIALS AND METHODS: Doppler echocardiography was made to evaluate diastolic function of the heart ventricles in 54 men with vibration disease. Mean exposure to local vibration was 22.4 years. The examinees were divided into two groups: 42 men (mean age 46.8 years) with vibration disease of the first degree- group 1; 12 men (mean age 51.2 years) of the first-second and second degree- group 2. RESULTS: In group 2 compared to group 1 there was a decrease of maximal velocity of early filling (MVEF) by 24.2%, early filling phase (EFP) by 27%, ratio MVEF/MVLF 1.17-fold, increase of isometric relation phase by 15.9%. For the right ventricle there was a decrease in MVEF by 15.4%, MVEF/MVLF 1.51-fold, prolongation of EFP by 20%. By these indices, the groups differed significantly in the diseases severity. Trimetasidine had a corrective effect on diastolic function of both left and right ventricles of the heart. CONCLUSION: Patients with vibration disease were diagnosed to have left and right ventricular diastolic dysfunction. PMID- 11494445 TI - [MR imaging in the diagnosis of rheumatoid arthritis]. AB - AIM: To study diagnostic potential of MR imaging (MRI) in rheumatoid arthritis (RA). MATERIAL AND METHODS: Clinical-radiation and MRI examinations of the joints were performed in 110 patients. RESULTS: MRI symptoms of RA were determined including accumulation of liquid in the joint cavity, degeneration of the joint cartilage, meniscus, ligaments, proliferation of synovial coating, destructive changes in bone epiphysis. CONCLUSION: Comparative analysis of x-ray and MRI results showed advantages of MRI in detection of inflammatory changes in soft tissue structures of the joints in RA. PMID- 11494446 TI - [New approaches to diagnosis and treatment of fibromyalgia in spinal osteochondrosis]. AB - AIM: To compare effectiveness of manual and bioresonance therapies for fibromyalgia (FM) in spinal osteochondrosis (SO). MATERIAL AND METHODS: The trial enrolled 60 FM patients with x-ray diagnosis of SO. In addition to routine clinical examination all the patients have undergone kinesthetic study with estimation of the muscular syndrome index, brain echoscopy, neurological examination, electropuncture diagnosis. Group 1 patients received manual therapy (MT) and point massage (PM); group 2 patients received MT, PM and bioresonance therapy (BRT). The treatment took 5-6 weeks. The examinations were made before the treatment and 1-1.5 months after it. RESULTS: The response was observed in both the groups, but in group 2 it occurred more frequently and earlier, was higher and longer. BRT produces no side effects, has no contraindications, acts on the body systemically. It is rather effective against symptoms of neurocirculatory dystonia frequently diagnosed in FM patients. PMID- 11494447 TI - [Differential diagnostic and prognostic value of serum cytokines (soluble CD30, soluble interleukin-2 receptor, interleukin-10 and interleukin-6) in Hodgkin's disease and lymphosarcomas]. AB - AIM: Determination of the importance of serum cytokines (sCD30, sIL-2R, IL-10, IL 6) for diagnosis, response to chemotherapy and remission in patients with lymphogranulomatosis and lymphosarcoma. MATERIAL AND METHODS: Cytokine concentrations were measured in 87 samples of serum (plasma) from 54 patients by ELISA. Diagnosis of Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) was made histologically in 24 and 30 patients, respectively. RESULTS: The threshold concentrations of sCD30 (and less specific sIL-2R) for HD and NHL patients allowed to estimate sensitivity to chemotherapy after the second course. The threshold concentration of IL-10 can distinguish HD from NHL. Changes in IL-6 concentrations were nonspecific. CONCLUSION: On the basis of the threshold concentrations of sCD30 and IL-10 we offer the scheme of lymphoma diagnosis and prediction of the disease sensitivity to chemotherapy which reduce the duration of lymphoma restaging. PMID- 11494448 TI - [Acute coronary syndrome: stages of diagnosis determining treatment policy]. PMID- 11494449 TI - [Diagnosis and approaches to therapy of emotional disorders in patients with infectious diseases]. AB - AIM: To examine psychological and emotional disorders in patients with infectious diseases, to specify indications for their pharmacological correction. MATERIAL AND METHODS: Clinicopsychological, clinicofunctional and laboratory tests were made to examine 30 patients with infectious mononucleosis (19 females and 11 males) and 30 patients with serous meningitis (16 females and 14 males) aged 16 35 admitted to hospital on the disease day 2-14. RESULTS: Shmishek's questionnaire revealed various types of personality accentuations with dominating hyperthymic (30%) and cyclothymic (20%). According to the data of the clinical scale SCL-90, the greatest number of cases with values over normal was in patients with serous meningitis. Beck's questionnaire revealed clinical depression in 12 patients (40%) with acute serous meningitis, subdepression in 14(46.7%) patients, severe depression in 6(20%) patients with infectious mononucleosis. In convalescence, emotional disorders persisted in 4 patients with serous meningitis. CONCLUSION: Affective disorders in the above patients require consultation of the psychiatrist to decide on psychopharmacotherapy inclusion in combined treatment of infectious diseases to prevent lingering course. PMID- 11494451 TI - [A case of timely diagnosis and surgical treatment of renovascular arterial hypertension in a woman patient with fibromuscular dysplasia of the renal artery and development of aneurysm]. PMID- 11494450 TI - [Diagnosis and differentiated treatment of secondary immunodeficiencies]. AB - AIM: To evaluate alterations in the immune system (IS) in patients with different forms of secondary immunodeficiency and design of differentiated programs of reestablishment of defective functions depending on pathogenetically important type of deficiency of immunocompetent cells. MATERIALS AND METHODS: Clinicoimmunological examination was made in 678 patients with complicated course of infectious-inflammatory diseases. Immunotropic medicines and physicochemical impacts were used in accordance with types of disorders in the system of immune homeostasis. RESULTS: There was a pathogenetic heterogeneity of IS disorders in complicated course of infectious-inflammatory diseases: generalized forms of infection (bacterial shock, sepsis) are in 75% of cases associated with deficiency of effector functions of peripheral blood polymorphonuclear leukocytes of the second-third degree, progressive fall in production of IgG immunoglobulins (42%), cellular-humoral immunodeficiency (92%). In lingering acute inflammatory diseases activation of phagocytosis occurred in 30%, IgG and/or IgM rise was in 50%, phagocytic function deficiency occurred in 48%, low production of immunoglobulins in 24%, humoral-cellular immunodeficiency in 62%. Purulent infection is associated with secondary cellular-humoral immunodeficiency, lowering of the immunoregulatory index (47%), phagocytic function deficiency (up to 35%), hyperproduction of IgM. Recurrent bacterial-viral diseases form in immunocompromised patients with T-lymphocytopenia (56%) and cellular-humoral immunodeficiency (30%). CONCLUSION: Protracted chronic inflammatory diseases are characterized by variability of changes in the immune systems. Combined types of disorders were found in 52% of the examinees. Pathogenetic heterogeneity of the disorders are determined by concomitant and previous diseases, occupational hazards and intoxication, environmental conditions, etc. CONCLUSION: Immunocorrective therapy in secondary immunodeficiency is conducted with allowances for pathogenetically essential types of disorders in the system of immune homeostasis, clinical variant of complication of inflammatory process under control of immunogram. PMID- 11494452 TI - [Myelodysplastic syndromes: current diagnosis and classification]. PMID- 11494454 TI - [Critical states in the practice of underwater work]. PMID- 11494453 TI - [Emergency treatment of acute respiratory disorders in patients with chronic obstructive lung disease and bronchial asthma]. PMID- 11494455 TI - [Methodological aspects of performing exercise tests used in combination with perfusion scintigraphy of the myocardium]. PMID- 11494458 TI - Altering records--it seemed like a good idea at the time. PMID- 11494457 TI - The sound of music. PMID- 11494456 TI - [Substantiation of increasing doses of statins in clinical practice]. PMID- 11494459 TI - How dental practitioners can benefit from the Internet. PMID- 11494460 TI - HIPAA & dentists: a new way of sharing and caring. PMID- 11494461 TI - Oral lichenoid reactions versus lichen planus. PMID- 11494462 TI - Marginal adaptation of three Class II composite restorative techniques in vitro. AB - The purpose of this study was to evaluate the marginal integrity of three different Class II restorative techniques in vitro. Mesioocclusal (MO) preparations (no bevels) 4 mm x 3 mm x 2 mm were made on 15 freshly extracted human premolars mounted in cold cure acrylic. The preparations were randomly divided into three groups of five: Group I: Direct technique Group II: Indirect single-visit technique Group III: Indirect laboratory technique Group II cavity preparations were duplicated using alginate impression material (Jeltrate Plus, DENTSPLY Caulk) and a polyvinylsiloxane die material (Mach II, Parkell). Group III preparations were duplicated using polyether impression material (Impregum, ESPE) and type IV die stone (G.C. America Inc., Fujirock). All restorations were fabricated with Tetric Ceram (Vivadent-Ivoclar) microhybrid composite according to manufacturers' recommendations. The inlays were cemented using a dual-cure bonding system (All Bond II, Bisco) and a resin-based cement (Variolink, Vivadent Ivoclar) and polished according to manufacturers' recommendations. The specimens were examined under an environmental scanning electron microscope (Electroscan). None of the samples was sectioned before examination. The width of the largest gap at the gingival margin, between the restorations and preparations, was recorded in microns. The average gap width for each group was the following: Group I: 63.4 +/- 9.06 microns Group II: 73.54 +/- 13.9 microns Group III: 89.58 +/- 18.5 microns Statistical analysis (one-way ANOVA) revealed that a significant difference exists among the three groups (p = 0.03). Students t-tests revealed a statistical significant difference between Groups I and III (p < 0.02), but did not prove significant between Groups I and II (p > 0.10) and Groups II and III (p > 0.05). PMID- 11494463 TI - Successfully managing a confrontation. PMID- 11494465 TI - NOHIC: reaching special-care patients with a wealth of information. PMID- 11494464 TI - Chew on this: is there a dominant side for chewing? AB - It is well known that individuals tend to favor either their left or their right hand. Medical professionals further acknowledge the existence of a dominant foot and eye. Whether we have a dominant side for chewing has not yet been established, but if so, does it correspond with our favored hand? Based on Decayed, Missing, and Filled (DMF) scores, researchers wonder whether the side of the mouth on which an individual predominantly chews in-fluences the amount of dental treatment a person has had or is in need of. This paper shows results of a survey of 112 people to determine if a pattern of DMF scores is predicative of a dominant side for chewing. In addition to a brief written survey, respondents underwent a complete oral and radiographic exam. Any decayed, missing, and filled surfaces were counted on both the right and left quadrants. These figures were then compared with the answers from the questionnaire to determine if any correlation could be made between DMF scores, age, gender, dominant hand, and eating and brushing habits. Results show that, in general, women believe they chew more on one side than the other, whereas men feel they chew on both sides equally. No statistically significant correlation was found between the hand that holds the fork, the side of the mouth chewed on, and the DMF score. This study did not confirm the existence of a dominant side of the mouth with respect to chewing. PMID- 11494466 TI - Drug interactions: COMT inhibitors. AB - To reduce the incidence of adverse drug interactions during dental treatment, always consult drug references when an unfamiliar drug is detected via the health history. It is crucial to understand the effects of a patient's preexisting disease and the medications used to treat it and their implications on dental therapy. PMID- 11494467 TI - Ameloblastic fibro-odontoma. AB - Ameloblastic fibro-odontoma is a benign, mixed odontogenic tumor most commonly encountered in the mandible of children or teenagers. Treatment of AFO is conservative and requires a long-term follow-up. Although some authors believe that ameloblastic fibroma, ameloblastic fibro-odontoma, and odontomas are extensions of the same disease process, they should be regarded as separate disease entities. PMID- 11494468 TI - The golden rule of dentistry. PMID- 11494469 TI - Invisalign: technology or mythology? PMID- 11494470 TI - The strategy. PMID- 11494471 TI - Practice management: then and now. PMID- 11494472 TI - The practice management revolution. PMID- 11494473 TI - HIV-positive patients. A duty to treat. PMID- 11494474 TI - From the patients' point of view. PMID- 11494476 TI - Bruxism and the bite. PMID- 11494477 TI - Subjugation of autonomy. PMID- 11494478 TI - Glenn Okihiro: a tireless fighter for patients' rights. PMID- 11494479 TI - [Sanitary assessment of currently available methods of drinking water disinfection (review)]. AB - Chemical and physical methods of drinking water disinfection do not meet the present day sanitary requirement in a number of essential indices. The disadvantages of routine methods make investigators search for new procedures generally based on the combined influence of two or several factors. Preliminary assessment of the existing procedures and those under development to disinfect drinking water indicates that physicochemical methods relating to a group of photo-oxidative and electrochemical methods, low-energy and pulse discharges in particular, will hold the greatest promise in future. PMID- 11494480 TI - [Study of carcinogenic hazards of smoking kiln waste fumes]. AB - The disperse phase of waste fumes forming during cold and hot fish smoking was studied by distributing the particles into 6 fractions and determining benz(a)pyrene (BP) in them. The level of BP in less than 2.5 mu was shown to be 85% whether smoking was hot or cold. The levels of 17 priority polycyclic aromatic hydrocarbons (PAH) were measured in three-dimensional fractions of fume particles. In all the fractions, phenanthrene and fluoranthene were of priority in weight. The proportion of BP in the total levels of PAH in waste fumes of cold and hot smokers was 0.69 and 0.74%, respectively and the carcinogenic activity was 46.42 and 49.0%. The investigations demonstrated that the waste fumes of smokers did not contain the particles that are PAH carriers. PMID- 11494481 TI - [Substantiation of an algorithm of sanitary measures to improve quality of drinking water]. AB - The authors analyzed different data available in the literature, reports of inspection of sanitary-and-epidemiological facilities and departments for ensuring the standard quality of water, and the results of long-term activities to eliminate the negative influence of water agent on morbidity. The protocols of epidemiological investigations of water-induced outbreaks of acute intestinal infections (AII) in the Republic of Byelarus in 1987-1997 were explored. The duration of outbreaks of water-borne AII in the country in these years was found to depend on the time of elaboration of an effective scheme of measures and on that of properly informative analysis of the established situation due to the quality of routine drinking water supply for the population. The more rapidly these measures are implemented, the shorter the outbreaks of AII are. An algorithm (schemes of laboratory control and measures) is substantiated and introduced for working out measures to eliminate the cause of water quality deterioration in the central water supply system. PMID- 11494482 TI - [Features of hormonal homeostasis under adverse industrial and ecologic conditions (a review)]. AB - The paper deals with changes in hormonal homeostasis under the long-term influence of industrial and ecological factors. It presents data available in the literature on adaptive and abnormal hormonal changes in workers exposed to noxious agents and in the residents of ecologically poor regions. Emphasis is laid on hormonal imbalance in the pathogenesis of different diseases, including occupational ones. PMID- 11494483 TI - [Results of activities of the State Sanitary and Epidemiological Service of the Russian Federation in 2000 and urgent tasks in the year 20001]. PMID- 11494484 TI - [Sanitary assessment of water quality in the city of Khiva]. AB - The paper deals with the 1994-1999 studies and hygienic assessments of the quality of water of the basis superficial water objects, point and shaft wells, tap water used by the population of the city of Khiva and its region. Sanitary features of the population's water use were found. The quality of tap water does not comply with the sanitary standards in the total hardness and sanitary and bacteriological indices. PMID- 11494485 TI - [Prediction in hygiene science and sanitary practice]. PMID- 11494486 TI - [Monitoring of microbial communities of the aqueous ecosystems]. PMID- 11494487 TI - [Health status of the residents in the area polluted by fluorine-containing discharges from the Tajik aluminium producing plant]. AB - The paper presents the results of investigations and follow-up monitoring to study the environment-human health relationships in the vicinity of an aluminium plant to substantiate a complex of preventive and health-promoting measures. Experience in planning and organizing the monitoring of environmental pollution and the health status of workers and the general population may be useful in doing the same work in the vicinity of other aluminium-making works. PMID- 11494489 TI - [Ergonomic approaches to optimizing the industrial "man-machine-environment" system]. PMID- 11494488 TI - [Diagnostic value of the determination of fatty acid spectrum of expired air condensate in the workers of coal stripping]. AB - The fatty acid spectrum of an expired air condensate was studied in the workers of the Kharanor coal stripping and in the dwellers of Chita (a control group). Regularities in the changes of the expired air fatty acid profile were assessed by the degree of exposure to coal dust and by the length of service. The findings indicated the relationship of profile changes with the degree of exposure to occupational noxious agents: the proportion of saturated fatty acids decreases much more significantly in workers who were directly exposed to the dust than in those were indirectly done. It is concluded that the fatty acid composition of an expired air condensate adequately reflects the changes occurring in the respiratory system upon exposure to dust. PMID- 11494490 TI - [Clinical and epidemiological diagnosis of chronic dust-induced bronchitis]. AB - A unified scheme for comprehensive interview of workers of a large metallurgic works who come in contact with occupational noxious agents that cause respiratory organs was used to select 300 workers (199 men and 101 women). They underwent a bicycle exercise test that involved the determination of forced vital capacity (FVC), forced expiratory volume within the first second (FEV1), peak expiratory air flow (PEAF), and the Tiffeneau index. Moreover, the flow/volume ratio was examined on a Flowscreen device (Erich Eger) and a test with a bronchodilator (ventoline, 200 mu/kg) was also made. The authors conducted 64 assays of eosinophilic cationic protein (ECP) and 64 assays of mucinic antigen (MA). Serum ECP and MA levels were measured by immunofluoresence and sandwich immunofluorescence, respectively. Screening functional indices obtained from the flow/volume and bronchodilator tests are sensitive in both manifestative and latent dust-induced bronchitis so they permit identification of latent and obvious airways obstruction. ECP is a highly reliable diagnostic criterion for the asthmatic form of chronic occupational bronchitis and MA is for its inflammatory form. PMID- 11494491 TI - [Effects of biologically active food additives on digestive function in experimental lead and chromium poisoning]. AB - Biologically active additives in the Multi-Green and Licorice foods were studied for effects on digestive function upon 100-day experimental intoxication with lead and chromium in a dose of 5 mg/kg. The metals were found to produce local irritating and inflammatory effects on the intestinal mucosa and a resorptive toxic effects on pancreatocytic function. Multi-Green and Licorice diminished the manifestations of adverse effects of lead and chromium on the functional status of digestive organs. PMID- 11494492 TI - [Assessment of health risks upon exposure to methylated mercury]. AB - Accumulation of a toxic agent (2-4-fold maximum allowable concentrations) was found in the fish caught in the polluted area of the water storage basin of the Angara river. The authors quantified a risk for major pathological syndromes in individuals exposed to mercury on fish ingestion and assayed biosubstrates for the substance. The integral risk of major pathological syndromes in the potentially mercury loaded persons increased by 62.1% with age and only by 17.1% in the control group. In the risk pattern there were risks for neurological diseases, arterial hypertension, coronary heart disease, borderline mental disorders, and endocrine diseases. PMID- 11494493 TI - [Monitoring of heavy metals in mushrooms and mushroom products in the Tomsk Region]. PMID- 11494494 TI - [Dynamics of some indices of immune defense and physical fitness in school children]. AB - Examinations of 22 young unarmed self-defence sportsmen and 18 untrained schoolchildren indicated that the indices of nonspecific immunity defence--serum complement and lysozyme--depended on a season rather than on a motor activity regimen. Changes in the concentrations of hemoglobin and plasma iron were also obviously seasonal. These indices were the least in spring and winter as compared to autumn and winter. Blood iron levels were higher in the young sportsmen than in the untrained schoolchildren at all stages of a follow-up. There was a direct relationship between direct and indirect parameters of iron exchange, physical fitness, and nonspecific protection factors. In all teenagers, the activity of serum lysozyme and complement was directly related to the plasma level of iron and the school children's physical fitness was closely correlated with plasma and blood cell iron concentrations. PMID- 11494495 TI - [Health-related requirements for the print design of illustrated texts in publications intended for children]. AB - The fact that there are essential differences in the volumes of texts for continuous reading in the belles-lettres and those for episodic reading (encyclopaedias, comic books, etc.) and marked differences in the area of the type pages intended for illustrations made it necessary to substantiate differential requirements for these groups of publications. Hygienic requirements were corrected by using the data obtained from 150 experiments covering 36 secondary-form pupils. Based on the well-known data on temporary visual fatigue thresholds (TVFT) and on the experimental findings, the authors defined the ratio of higher TVFT to the actual time of reading the texts of different volumes. A complex of print design indices to be hygienically regulated varies with the multiplicity that is in fact a safety factor. PMID- 11494496 TI - [Social and sanitary factors and children's health]. AB - The paper shows how a complex of social and sanitary factors contributes to health formation (physical development, morbidity, premorbid conditions) in children aged 5-6 years. PMID- 11494497 TI - [Anthropometric characteristics of rural school children 8-13 years of age living in radioactive pollution areas]. AB - The paper presents the results of a study of age- and sex-specific features of physical development in schoolchildren aged 8 to 13 years who reside in the rural areas exposed to radioisotopes. Changes in their body weight, height, and weight height ratio are analyzed. PMID- 11494498 TI - [Principles of and criteria for sanitary standardization of antitumor cytostatics in the water of aqueous objects]. AB - The list of sanitary standards with the mark "absent" should include antitumor cytostatics as the drugs satisfying the following criteria: extreme hazard and toxicity, late adverse effects, primarily mutagenic and carcinogenic properties; lack of highly sensitive methods for their determination in water; lack of 100% efficiency methods for sewage purification. A complex of protective measures is justified and sanitary recommendations are given to protect the aqueous environment during cytostatic production. PMID- 11494499 TI - [Experimental-hygienic assessment of 2 binary mixtures: lead-copper and lead zinc]. AB - The paper presents the results of experiments to examine the effects of zinc and copper on some indices of the total toxic and specific action of lead. Experiments were conducted on non-inbred albino rats intraperitoneally given the acetates of lead and zinc, as well as copper sulfate alone and in combination with the acetates. Antagonism is a determining type of combined effects of the binary mixtures lead-copper and lead-zinc upon acute and subacute exposures. PMID- 11494500 TI - [Oncologic hygienic aspects of regulation of benz(a)pyrene in food products]. AB - The paper presents a methodological scheme for regulating the acceptable levels of benz(a)pyrene in the foodstuffs is presented. It also gives a list of the acceptable levels of this carcinogen in the foodstuffs whose specific values depend on its daily dosage and the pattern of a standard diet for the Ukraine's population. PMID- 11494501 TI - [Sanitary assessment of building materials containing industrial waste]. PMID- 11494502 TI - [Evaluation of the validity of the results of sanitary-microbiological analysis of water]. PMID- 11494504 TI - [Man-made environmental air pollution and children's health]. AB - Ambient air pollution and morbidity were studied by the visits of children under 7 years to a doctor. A total of 950 children living in the areas that are notable for the level and pattern of ambient air pollution were examined. The greater pollution of development lands is, the higher general morbidity and childhood morbidity of respiratory nervous, sensory, skin, and subcutaneous fat diseases are. Ambient air chlorinated and aromatic hydrocarbons may cause a high incidence of biliary and hepatic diseases and chlorinated hydrocarbons in children. Equations have been derived to predict childhood morbidity due to the level and pattern of air pollution. PMID- 11494503 TI - [Use of gas and liquid chromatography methods in studying the metabolism of organic compounds (review)]. PMID- 11494505 TI - [Dissecting the structure and function of the extracellular hyaluronan-rich matrix by gene manipulation]. PMID- 11494506 TI - [Functional analysis of proteoglycans by knockout technology]. PMID- 11494507 TI - [Heparan sulfate proteoglycans: Drosophila genetics and biochemistry provide a new insight into the regulation of their functions]. PMID- 11494508 TI - [Biosynthetic mechanism of sulfated glycosaminoglycans]. PMID- 11494509 TI - [Proteoglycans in the development, lesion, and aging of the brain]. PMID- 11494510 TI - [Heparanase: heparan sulfate specific endo-beta-D-glucuronidase]. PMID- 11494511 TI - [Functional roles of GlcNAc N-deacetylase/GlcN N-sulfotransferases in the biosynthesis of heparan sulfate/heparin]. PMID- 11494512 TI - Protein expression and cell organelle behavior in spermatogenic cells. AB - Spermatogenic cells stage-specifically produce a wide variety of proteins during spermatogenesis, wherein protein expression is coordinated with cell organelle behavior. It has been shown that the Golgi apparatus and the endoplasmic reticulum (ER) are uniquely coordinated with the expression of an immunoglobulin super-family protein, flagellar plasma membrane MC31 (MC31/CE9), and a molecular chaperone, calmegin, respectively. When the Golgi apparatus begins to generate sperm components in the primary spermatocytes, it actively engages in producing proteins for the acrosome in round spermatids and for the flagellum in elongating spermatids. Structurally, the Golgi apparatus is reduced in size during meiotic division, moves from the apical to the basal region (cytoplasmic lobe) when spermatids differentiate from round to elongating phase, and then collapses in the late maturation phase. The ER is distributed uniformly over the entire cytoplasm of spermatocytes and round spermatids, and then moves distally toward the cytoplasmic lobe along the bundles of microtubule, called the manchette, in elongating spermatids. The ER is resorbed into the radial body in late maturation spermatids. MC31/CE9 expresses strong immunostaining twice on the Golgi apparatus during spermatogenesis, first in early pachytene spermatocytes and then in early elongating spermatids. Calmegin expression exactly parallels ER behavior. This mini-review focuses on the unique relationships in spermatogenic cells, particularly those between protein expression and cell organelle behavior. PMID- 11494513 TI - [Morphology and significance of the left portal vein in human]. AB - The aberrant course of a left gastric vein directly entering the liver has been reported as the left portal vein in a few cases. The authors adopted the name of 'left portal vein' for this vein and gave a brief historical summary of the left portal vein and its significance. The left gastric vein does not enter the gastropancreatic ligament from the lesser curvature, but the vein runs toward the liver alongside the hepatic ramus of vagus nerve in the hepatogastric ligament. The vein either joins the branch of the portal vein or supplies a small area of the liver after reaching the left side of the hepatic hilus. The left portal vein has already been described as the term of 'V. portae sinistra' in the bird. Many vascular vessels are found in the hepatogastric ligament. In this ligament the hepatic ramus of vagus nerve always runs toward the liver and the artery exists at a frequency of 30%. Either the left hepatic artery arising from a left gastric artery runs alongside the nerve in the ligament at a frequency of 20% or the accessory left gastric artery arising from a hepatic artery runs in a reverse direction at a frequency of 10%. Both arteries do not coexist. Moreover, the left gastric vein exits in the hepatogastric ligament in several cases and enters the liver as the left portal vein. Besides, an anastomosis of a portal vein with the renal vein rarely exists in front of the cardiac region of the ligament in cirrhosis. The aberrant left gastric vein is derived from the hepatic afferent vein through the hepatogastric ligament and exists as the left portal vein. PMID- 11494514 TI - Ureter duplication and polar renal arteries in the same case. AB - During cadaver dissection for student education in the anatomy laboratory of Gazi University Medical School, anomalies of the upper and lower polar artery were observed on the left kidney besides bilateral ureter duplication in a 65 year old male cadaver. On the right side the duplication was incomplete, starting from two separate renal pelvises, joining at the medial border of psoas major muscle and terminating as a single specify at the level of urine bladder. On the left side the duplication was complete. The ureters started from two different renal pelvises and terminated separately. In addition, two polar arteries were observed on the left kidney besides the normal renal artery in the same case. The upper polar artery arose just at the beginning of left renal artery and the lower one arose directly from the lateral wall of abdominal aorta. Due to the surgical and radiological significance of the variations they presented, we compared them with the other studies on this subject. PMID- 11494515 TI - Gap junctional communication between the satellite cells of rat dorsal root ganglia. AB - Many studies have described the ultrastructure of the dorsal root ganglia in various embryonic and adult animals, but in spite of the efforts of many investigators the functional role of the satellite cells in this tissue is not clearly understood. In this study, we discuss the function of this cell type based on the concept of cell-to-cell interaction through gap junctions. Five male 60 day-old Wistar strain rats were used. All animals were anesthetized with pentobarbital and perfused with glutaraldehyde fixative, then the dorsal root ganglia in levels L4, L5 and L6 were taken from each rat. After postosmication, the specimens were prepared for observation by transmission electron microscopy. All nerve cells were completely surrounded by satellite cell cytoplasmic expansions. The boundaries between adjacent nerve cells and satellite cells were complicated due to the presence of perikaryal projections of nerve cells. Gap junctions which showed the typical trilamellar structure of plasma membranes were found mainly between satellite cell processes belonging to the same nerve cell. On the other hand, some gap junctions were found between the satellite cell projections belonging to different nerve cells. The size of the gap junctions ranged from 300 to 400 nm. No gap junctions were associated with the plasma membrane of any nerve cell. In conclusion, only satellite cells can share free transcellular exchange of cytoplasmic molecules such as ions, amino acids, sugars and several second messengers including cAMP and inositol 1,4,5-triphosphate by way of gap junctions in dorsal root ganglia. PMID- 11494516 TI - Distribution of preganglionic terminals in the cervical sympathetic ganglia detected by the expression of c-Fos like protein after electric stimulation of the ventral root. AB - To determine the segmental relationship between the upper thoracic spinal cord and cervical sympathetic ganglia, we observed the distribution pattern of postganglionic cells which expressed c-Fos like protein, one of the products of immediate early genes, after electrical stimulation of ventral roots at the T1-T3 spinal segments. We recognized a clear segmental arrangement of postganglionic cells in the stellate ganglion along its rostrocaudal direction corresponding to the segmental arrangement of preganglionic neurons in the spinal cord. That is, postganglionic neurons which expressed c-Fos like protein after stimulation of the T1 ventral root were distributed in the middle region of the stellate ganglion in the rostrocaudal direction. The c-Fos like protein-positive neurons after stimulation of the T2 ventral root were distributed in a more caudal region of the stellate ganglion than after T1 ventral root stimulation. C-Fos like protein-positive neurons after stimulation of the T3 ventral root were mainly situated in a more caudal region of the stellate ganglion than after T2 ventral root stimulation. There was, however, no segmental relationship between the upper thoracic levels of the spinal cord and superior cervical ganglion in the rostrocaudal direction. These results indicate that the segmental innervation of the upper thoracic spinal cord exists in the stellate ganglion, but not in the superior cervical ganglion. PMID- 11494517 TI - [Innervation of metacarpophalangeal joint and distal interphalangeal joint: an anatomical and histological study]. AB - Using 131 fingers for the metacarpophalangeal (MP) joint and 124 fingers for the distal interphalangeal (DIP) joint obtained from 30 hands of human cadavers, the innervation of the MP and DIP joints was investigated anatomically in detail. Two articular branches developing from a dorsal branch of the ulnar nerve and a superficial branch of the radial nerve, and entering the dorsal side of the MP joint from the ulnar side and radial side, respectively, were found in every finger. In addition, articular branches from the deep branch of the ulnar nerve were found in every middle, ring and little finger. However, articular branches from the proper palmar digital nerve were found to exist in 62.7% of the fingers. Articular branches developing from the proper palmar digital nerve and going towards the DIP joint were found in every case, and 97.3% of 244 branches developed directly from the proper palmar digital nerve, while some of the remaining branches were from a dorsal branch of the proper palmar digital nerve and others developed neural loop penetration. Articular branches ran parallel to the distal transverse artery and entered the joint, and some of them went towards the volar plate, dorsal joint capsule, and tendon sheath. Many nerve endings existed in the surface layer of the articular capsule and arthrosynovial membranes in the form of Pacinian corpuscles and corpuscles of Ruffini. The existence of a neural loop in the finger should be taken notice of during somatoscopy in patients with neurovascular symptoms in the fingertip or surgical operation on a vascular pedicular island flap. Further, it was suggested that injury of the articular branch of the DIP joint could induce Heberden nodes due to its anatomical characteristics. PMID- 11494518 TI - A case of hypoplasia of the left hepatic lobe. AB - We report a case of hypoplasia of the left hepatic lobe in a 90-year-old woman. The left lobe was composed of three parts, central, anterior and lateral. The central part was equivalent to the hepatic hilum and consisted of 1/3 of a circle with a radius of about 30 mm and a maximum thickness of 8 mm. The anterior part had an elliptical shape of 65 mm x 33 mm, 21 mm thick. The lateral part had an elongated elliptical shape of 67 x 21 mm, also 16 mm thick. Sublobation was noted in the caudate and quadrate lobes. The caudate lobe was divided into two parts, and the quadrate lobe into three parts. The right lobe was bell-shaped with the top turned toward the diaphragm. The transverse diameter at the base was 108 mm, the antero-posterior diameter was 134 mm, and the distance from the bottom to the top was 95 mm. A deep groove paralleled the anterior rim at the base surface, where the gallbladder was in contact with the liver. No anomalous distribution of blood vessels or biliary canals was found. No atrophic feature was histologically recognized in any lobe. PMID- 11494519 TI - [Anterior turbinectomy: experimental and clinical study]. AB - INTRODUCTION: Minimal cross-sectional areas situated at the anterior end of the inferior turbinate have the most influence on nasal patency. To improve persistent nasal obstruction, 2 types of inferior turbinectomies--conventional complete resection including the anterior and posterior mucosa and anterior resection of the anterior mucosa--were conducted and preoperative and postoperative nasal patency and stuffy sensation were compared and analyze. OBJECTS AND METHODS: Cases undergoing 2 types of turbinectomies between July 1997 and March 2000 numbered 63--conventional in 32 (64 sides) and anterior in 31 (62 sides). Anterior, posterior, and total nasal volume and minimal cross-sectional area were evaluated pre- and postoperatively using an acoustic rhinometer. The stuffy sensation was similarly evaluated by the visual analog scale (VAS). RESULTS: All nasal volumes showed postoperative increase under conventional treatments. Anterior and posterior volumes significantly increased under anterior treatment. Comparing the results of postoperative volume posterior volume was significantly larger under conventional treatment than under anterior treatment. Stuffy sensation significantly improved by VAS in both types of turbinectomy, with no significant difference between them. CONCLUSION: Anterior treatment to solely increase nasal volume at the minimal cross sectional area showed equal postoperative improvement in VAS under conventional treatment. PMID- 11494520 TI - [Medical cost analysis of neoadjuvant chemotherapy for advanced tongue and oropharyngeal cancer]. AB - We performed a medical cost analysis on the use of neoadjuvant chemotherapy (NAC) in multi-modal treatments for advanced tongue and oropharyngeal cancer. Twenty two patients with stage III and IV squamous cell carcinoma of the tongue and oropharynx were enrolled in this study. Two courses of NAC with CDDP and 5-FU followed by radiotherapy were performed in 13 patients who showed a partial response or a complete response after the first course of NAC. The doses of radiation were 60 Gy for 4 patients (rad. group), and 72 Gy for 9 patients who received hyperfractionated radiotherapy with simultaneous Carboplatin (HF rad. group). Radical surgery was performed in 9 patients who did not respond to NAC (surg. group). The mean duration of hospital stay was 89.3 days for the rad. group, 92.0 days for the HF rad. group, and 113.3 days for the surg. group. The mean medical cost was 238,700 points for the rad. group, 264,846 points for the HF rad. group, and 459,468 points for the surg. group. The mean amounts and percentages of cost for NAC were 39,473 points and 16.1% for the rad. group, 44,802 points and 16.9% for the HF rad. group, and 23,451 points and 5.1% for the surg. group. The mean amounts of cost for NAC including examination costs and nursing costs for chemotherapy were 130,196 points and 54.5% for the rad. group, 150,046 points and 55.7% for the HF rad. group, and 113,839 points and 24.8% for the surg. group. The cost of NAC accounted for half of the total cost of chemo radiation treatment for functional preservation. The duration of the hospital stay was prolonged by NAC, which accounted for a quarter of the total cost for the surg. group. Survival benefits are considered in the analysis of treatment cost-effectiveness for head and neck cancers. In the future, QOL, which is defined as the utility achieved as a result of multi-modal treatments, should also be analyzed along with survival benefits. Cost-utility analyses should include quality-adjusted life years, based upon the cost identification results of our analysis for NAC, to evaluate the efficacy of NAC in multi-modal treatments for head and neck cancers. PMID- 11494521 TI - [Nasal allergies in Kushiro]. AB - A clinical study of 107 patients with nasal allergies who were treated at Kushiro General Hospital between April 1998 and March 2000 was performed. Radioallergosorbent tests (RAST), X-rays, and nasal smears for the detection of eosinophilia were performed to obtain a diagnosis of nasal allergy. The patients (50 males, 57 females) ranged in age from 3 to 71 years. The RAST was positive for timothy in 22.4% of the patients, 14.0% for birch and 12.1% for mugwort. The most common pollinosis allergen in the Kushiro area was grass pollen. Other pollinosis allergens were birch pollen and mugwort pollen. We measured the daily count of dispersed birch pollen and timothy pollen in Kushiro and Sapporo. Birch pollen and timothy pollen was dispersed earlier in Kushiro than in Sapporo. Nasal allergies in the Kushiro area appear to be related to local characteristics, such as climate and geographical features. PMID- 11494522 TI - [Changes in otoacoustic emission during the glycerol test in the ears of patients with Meniere's disease]. AB - Monitoring of cochlear function is one proposed clinical application of otoacoustic emission (OAE). To determine the clinical utility of OAE in monitoring, we studied changes in OAE during the glycerol test used to diagnose endolymphatic hydrops. Transiently evoked OAE (TEOAE) and distortion product OAE (DPOAE) were measured before and 3 hours after oral administration of glycerol in 22 ears of patients with Meniere's disease. The positive result in the glycerol test was observed in 11 of 22 ears. TEOAE was recorded using a nonlinear click stimulus, and total echo power (TEP) of the whole response and filtered echo power (FEP) of its mid-frequency (1000-2000 Hz) component were used as TEOAE parameters. The growth functions of DPOAE were recorded at 3 F2 frequencies- 1000, 1500, and 2000 Hz. Changes in the growth function were determined based on changes in 2 parameters--maximum level and detection threshold of DPOAE. When changes in these 4 parameters exceeded 2 standard deviations above mean test retest variability determined from OAE data for 17 normal ears, they were considered significant. Significant changes indicating improved cochlear function in 4 OAE parameters or the appearance of OAE during the glycerol test were judged positive for OAE. Regardless of glycerol test results, positive findings were observed in both OAEs whenever a pure-tone threshold at 1000 or 2000 Hz improved. However, even in ears for which pure-tone thresholds at these frequencies did not improve, both OAEs frequently exhibited positive findings. In all ears with a positive result in the glycerol test, positive findings were observed in DPOAE, but not in TEOAE. Of the 2 parameters of TEOAE, FEP always exhibited a positive finding in ears with a positive TEP finding and FEP positivity was higher than that of TEP. In DPOAE, no cases showed a positive finding only at F2 = 2000 Hz, and DPOAE at F2 = 1000 Hz could not be detected in 4 ears due to high noise floor. Overall, positive findings were obtained in 14 ears (63.9%) for TEOAE and 20 ears (90.9%) for DPOAE. In only 1 ear were no positive findings observed in either OAE. The sensitivity of OAE measurement as a diagnostic test for endolymphatic hydrops was higher than that of pure-tone audiometry. Of the 2 OAEs, test sensitivity was much higher for DPOAE than for TEOAE. These results show that OAE measurement detects changes in cochlear function during the glycerol test more sensitively than pure-tone audiometry. They also suggest that FEP is a better indicator than TEP as a parameter of TEOAE, and that DPOAE measurement at F2 = 1000 and 1500 Hz is sufficient for practical clinical use. However, of the 2 OAEs, DPOAE was considered more appropriate than TEOAE for monitoring during the glycerol test because of its high sensitivity in detection of changes in cochlear function. In conclusion, clinical use of OAE, especially DPOAE, as a test complementary to pure-tone audiometry during the glycerol test is very useful and will improve the diagnosis of endolymphatic hydrops. PMID- 11494523 TI - [Multivariate analysis using quantitative methods to evaluate the subjective loudness of tinnitus]. AB - Factors affecting the subjective loudness of tinnitus were analyzed using quantitative methods. One hundred and eighty-four patients with unilateral tinnitus were studied. Major factors affecting the subjective loudness of tinnitus were found to be subjective tinnitus pitch, annoyance, diagnosis, pitch matched frequency, insomnia, age, tinnitus loudness level in dB HL, the duration of tinnitus and the tinnitus loudness level in dB SL at the first axis. Tinnitus pitch and pitch-matched frequency strongly affected the loudness of the tinnitus; high-pitch tinnitus and middle and low frequencies were associated with the subjective loudness of the tinnitus. In contrast, low-pitch tinnitus and a high frequency were associated with the subjective softness of the tinnitus. Thus, tinnitus pitch and pitch-matched frequency are inversely related. Sudden deafness and dizziness were associated with subjective softness of tinnitus, while hearing loss in elderly patients was associated with subjective loudness of tinnitus. Our results suggest that the level of annoyance from tinnitus and the subjective loudness of tinnitus are closely related. The relation between these factors should be considered when evaluating patients with tinnitus. PMID- 11494524 TI - [A study on the clinical features of pulmonary tuberculosis in elderly patients]. AB - The clinical findings of pulmonary tuberculosis in elderly patients aged 75 years of age or older (elderly group) were compared with the middle-aged patients aged between 45 and 54 years of age (middle-aged group) and the young patients aged 34 years of age or younger (young group). The elderly patients who died in hospital were also compared with the elderly patients who survived. Study subjects were culture-positive pulmonary tuberculosis patients who were discharged from our hospital from December 1996 to November 1998. There were 79 patients in the elderly group, 95 in the middle-aged group, and 88 in the young group. The results were as follows. 1) The male/female ratio was significantly lower in the elderly group (1.9:1) than the middle-aged group (6.9:1). 2) Complication was noted more frequently in the elderly group. However, diabetes mellitus was noted less frequently in the elderly group (12.7%) than the middle-aged group (28.4%). 3) The frequency of cavitation was lower in the elderly group (59.5%) than the middle-aged group (87.4%). 4) The fever over 38 degrees C was noted less frequently in the elderly group (17.7%) than the other groups, while the frequency of the fever over 37 degrees C showed no significant difference between the elderly group and the other groups. 5) The frequencies of hypoalbuminemia and appetite loss were higher in the elderly group than the other groups. 6) The elderly group showed high mortality rate of 31.6%. The complication with cerebrovascular disease was noted significantly higher in the patients who died in hospital than those who survived. The frequency of widespread infiltrates, fever over 38 degrees C, neutrophilia, hypoalbuminemia and appetite loss were all significantly higher in the patients who died in hospital while their sputa were still positive on culture than those who survived. Our study clearly showed the features of elderly pulmonary tuberculosis patients in comparison with middle aged patients and young patients. These features are very important to suspect the diagnosis of pulmonary tuberculosis in elderly patients with some atypical manifestation. Our study also suggests that the delay in diagnosing tuberculosis causes more frequently the patients' deterioration and death in elderly patients than in middle-aged patients and young patients. PMID- 11494525 TI - [A study on family infection of tuberculosis]. AB - The guideline for contacts examination and chemoprophylaxis for tuberculosis was revised in 1993. Secondary cases and chemoprophylaxis cases found by the contacts examination of family members were analyzed in Sapporo City. The average age of index cases and of secondary cases has become higher. Many secondary cases were found among spouse of the index case and children of male index case. Chemoprophylaxis cases were found mostly among children of male or female index cases and grandchildren of male index cases. The results of family contacts examination carried out during the period from 1994 to 1999 were compared with those from 1987 to 1992. The incidence of family infection of tuberculosis and the number of newly infected persons and of secondary cases have decreased in the latter period. The time interval from the registration of the index case to the detection of secondary cases has become shorter, and the ratio of sputum smear positive cases among secondary cases has also decreased. Male to female ratio of the index cases was 2:1, that of secondary cases was 3:5, and that of children indicated chemoprophylaxis was 4:5. All secondary cases found by the contacts examination carried out within 6 months after the previous examination were sputum smear negative for tubercle bacilli, and there were a few smear positive cases among secondary cases who were found by the examination with longer interval with the previous examination. Such cases with longer interval from the last periodic examination was found often among younger cases below 40 years of age. Most smear negative tuberculosis cases were found by the family contacts examination, while most smear positive cases were found by symptomatic visit to doctors. PMID- 11494527 TI - [A case of tuberculous endometritis detected by cytology of mass screening for gynecologic cancer]. AB - A 55-year-old infertile woman was referred to our clinic for further investigation on extragenital tuberculosis, as tuberculous endometritis was strongly suspected by cytology of her vaginal smear carried out on the occasion of the mass examination for gynecologic cancer screening. Her vaginal smear revealed epithelioid cell clusters which are characteristic for tuberculosis, and cultures of her vaginal discharge were positive for M. tuberculosis consecutively. Moreover, she was exposed for tuberculosis infection from her father who died of active pulmonary tuberculosis when she was ten years old. Her tuberculin test was strongly positive, and her chest radiography showed no abnormality, but a small nodular shadow evaluated as primary focus of tuberculosis located beneath the pleura of the right lower lung field was confirmed by chest CT. In addition, calcification of her para-aortic abdominal lymphnode was detected by simple abdominal X-ray. Based on these data, she was diagnosed as tuberculous endometritis via abdominal cavity, and three antituberculous drugs, namely RFP, INH and EB, were administrated. The mycobacterial cultures of vaginal discharge converted to negative, and chemotherapy was terminated after 9 months treatment. A risk factor leading to the onset of gynecologic tuberculosis, in this case was an exposure to infection from her father. In order to evaluate risk factors relating to the development of gynecologic tuberculosis, bibliographic studies were made on 19 cases of tuberculous endometrites reported recently in Japan regarding their age, its pathogenesis and immuno-suppressive conditions, and the summarized results were as follows. 1. approximately 80% of them were elderly, namely 79% were above 50 years, 63% above 60 years, and 26% above 70 years. 2. 50% of them were caused by endogenous reactivation. 3. 25% of them were immuno-compromised host. It can be concluded that more than 70% of the patients with tuberculous endometritis had risk factors on the host side to develop tuberculosis. PMID- 11494526 TI - [A study on resistance of Mycobacterium tuberculosis to four first-line anti tuberculosis drugs in Japan: comparison of results in the local facilities and in the reference laboratory, in 1997]. AB - Five years after the last survey of drug-resistant tuberculosis in Japan, a nationwide survey was conducted by the Tuberculosis Research Committee (Ryoken). A total of 78 hospitals in various districts of Japan participated in this cooperative study. Each collaborating laboratory sent all the isolated mycobacterial cultures during June 1 to November 30, 1997 to the Research Institute of Tuberculosis (RIT), which is one of the Supranational Reference Laboratories of the WHO/IUATLD Global Project on Anti-tuberculosis Drug Resistance Surveillance. At RIT identification and drug susceptibility of Mycobacterium tuberculosis isolates were reexamined. The RIT received a total of 2,167 cultures. Among them, 523 cultures were excluded from further examinations because of various reasons, such as growth of mycobacteria other than tubercle bacilli (MOTT, 453), mixed cultures of M. tuberculosis and MOTT (16), and contamination or non-viability (54). Thus drug susceptibility test results were available for 1,644 cultures, including 47 from foreign-born people. In the local laboratories, the absolute concentration method using 1% Ogawa egg slant (standard method, 26 hospitals), its modified methods using a 48-well plate (Microtiter method, 29 hospitals) and a 16-well plate (Well-pack method, 7 hospitals), combination of above 2 or 3 methods (13 hospitals), and other method (3 hospitals) were used for drug susceptibility testing, and the proportion method using 1% Ogawa egg slant was used in the RIT. The results in the local laboratories were compared with those in the RIT. A high concordance rate (over 90%) was seen in the testing for 1 microgram/ml of isoniazid (INH), rifampin (RFP) and streptomycin (SM), but the rate was lower (under 90%) in the testing for 0.1 microgram/ml of INH and ethambutol (EMB). However, there was no significant difference in the concordance rates according to the test drugs among methods for drug susceptibility testing used in the local laboratories. Median concordance rates between the results with the standard method, Microtiter method and Well-pack method in the local laboratories, and those in the RIT were 95.9%, 93.2% and 96.4% respectively. Relatively lower concordance rates were seen in the laboratories using the Microtiter method related to high overestimation rates (median overestimation rate of 5.3%), compared with 1.2% and 2.3% in the laboratories using the standard method and Well-pack method, respectively. However, relatively lower concordance rates (less than 90%) were seen in the laboratories using any of the three methods, indicating that there are variations among facilities. Part of the results concerning the resistance patterns to four first-line anti-tuberculosis drugs were reported elsewhere. PMID- 11494528 TI - [A case of tuberculous peritonitis diagnosed by a direct smear of ascitic fluid complicated with an active pulmonary tuberculosis and intestinal tuberculosis]. AB - We sometimes encounter difficulties in differentiating tuberculous peritonitis from other inflammatory disorders or ascites due to carcinomatous peritonitis. Acid-fast bacilli are very rarely detected in ascites. In this study, we reported a case of tuberculous peritonitis accompanied with active pulmonary tuberculosis in which acid-fast bacilli were detected in ascites. The patient was a 37-year old single man who had been admitted to our hospital on February 28, 2000, because acid-fast bacilli were detected in sputum, faces and ascites by a direct smear. He had a lower abdominal distention and pain. His serum CA 125 level was high, 121 U/ml. Abdominal ultrasonography showed marked ascites in Douglas pouch. However adenosine deaminase level was not high in his ascites. During treatment by the combination chemotherapy with INH, RFP, EB, and PZA, serum CA 125 level was decreased. PMID- 11494529 TI - [A case of Mycobacterium intracellulare lung disease occurred in a middle-aged man who complained hemoptysis with normal chest roentogenogram]. AB - A 45-year-old man visited our clinic because of intermittent bloody sputum. The chest roentogenogram was normal, but the high-resolution computed tomography (HRCT) showed very small nodules and bronchiolitis adjacent to pleura in the upper right lung field. The bronchoscopic examination revealed blood-streaked bronchial secretion in the right upper lobe bronchus, and the cultures of the sputa and the bronchial washing specimen showed acid-fast bacilli identified as Mycobacterium intracellulare by DNA-DNA hybridization (DDH) method. This case was diagnosed as Mycobacterium intracellulare lung disease. The patient received isoniazid, levofloxacin, and clarithromycin for three years without clinical and bacteriological improvement. His hemoptysis and the number of colonies recovered from sputum cultures decreased without any medication later. The serial chest roentogenograms and HRCTs have showed no changes for 6 years after the diagnosis. This case may show some clues to elucidate the mechanism of the onset of Mycobacterium intracellulare lung disease without predisposing conditions. PMID- 11494531 TI - [Recent progress and prospective view of chronic hepatitis C research]. AB - Hepatitis C is a major public health problem because of the high incidence of its related hepatocellular carcinoma. With the progress in molecular biology, the mechanisms of persistent infection, chronic inflammation, and hepatocarcinogenesis have been described in terms of virus, host, and virus-cell interactions. On the other hand, clinically, some recent studies using a large number of subjects with long-term observation after interferon therapy showed that improving hepatic inflammation might be associated with regression or retardation of fibrosis. However, current therapy for hepatitis C, although effective in some patients, is problematic even though the efficacy of combination therapy, interferon plus ribavirin, or PEG-interferon has been reported. Here we review the progress and discuss the prospective view of hepatitis C virus research from a point of view of both basic and clinical aspects. PMID- 11494532 TI - [Involvement of TAP2 and LMP7 gene polymorphisms in HCV infection]. AB - We aimed to study whether TAP and LMP polymorphisms could influence the severity of liver disease or the response to IFN treatment in patients with chronic HCV infection. TAPZ*0103 gene frequencies in carriers with normal ALT was significantly higher than that in CLD patients. As for the results of IFN responses, LMP7-K gene frequency in sustained-responders was higher than that in non-responders. Multivariate analysis revealed that LMP7-K and HCV RNA quantity were independent factors influencing the outcome of IFN therapy. Furthermore, among patients with a low viral load, the LMP7-K positive patients had a significantly higher ratio of sustained response compared to those without LMP7 K. The TAP2 polymorphism may be closely associated with low hepatitis activity, whereas the LMP7 polymorphism influences the efficacy of IFN treatment and can be a useful predictive parameter in HCV patients with a low viral load. PMID- 11494533 TI - [HCV proteins suppress translation]. AB - Viruses influence host cell protein synthesis in various ways. There are many reports that viral infections inhibit host cell translation, in a process called 'translational shutoff'. In most cases, viral translational shutoff gives an advantage to virus survival because host cell translation is suppressed and the translational machinery is used to translate viral RNA instead. However, there are few reports on the effects of hepatitis C virus (HCV) infection on protein synthesis, because of the lack of a reproducible tissue culture system for HCV. We found that HCV also have the effect of translational inhibition. This novel function may help HCV survival. PMID- 11494534 TI - [Th1/Th2 imbalance in HCV-related liver cirrhosis]. AB - The mechanism by which Hepatitis C virus(HCV) infection promotes the development of hepatocellular carcinoma(HCC) is not known exactly. HCV related HCC occurs frequency in the patients with cirrhosis. There have been reports indicating that Th2-type cytokines down-regulated antitumor immunity, and the activation of type 1 T cell responses produced antitumor immunity. We thought Th1/Th2 imbalance in HCV-related liver cirrhosis might be closely related to the development of HCC. In this study, therefore, we investigated the Th1/Th2 balance at the single lymphocyte level of the patients with HCV-related liver cirrhosis and compared with normal controls by using flow cytometry. Th1-type cytokines(IFN-gamma, IL-2) production was significantly decreased in patients with cirrhosis, whereas Th2 type cytokine production(IL-10) was increased. These suggest Th1/Th2 imbalance in HCV-related cirrhosis would decrease the antitumor immunity and its improvement might present the protective effect from HCC. PMID- 11494535 TI - [HCV core gene transgenic mouse as a model for viral hepatocarcinogenesis]. AB - Overwhelming lines of epidemiological evidence have indicated that chronic infection with hepatitis C virus(HCV) poses a major risk toward development of hepatocellular carcinoma(HCC). In the pathogenesis of HCC associated with HCV, it remains controversial whether these hepatitis viruses play a direct role or merely serve for an indirect role. By virtue of transgenic mouse established by us, it has become evident hat the core protein of HCV has an oncogenic potential. The findings in our studies indicate that HCV is directly involved in hepatocarcinogenesis, albeit other factors such as continued cell death and regeneration associated with chronic hepatitis may play a role, as well. Combined, our results suggest that there would be a mechanism for the development of HCC in persistent infection with hepatitis C virus that is distinct from those for the other cancers. Similar to the pathogenesis of other malignancies represented by colorectal cancer, the accumulation of a set of genetic aberrations may also be necessary for a multi-stage development of HCC. However, HCV core protein, to which an oncogenic potential is ascribed, may allow some of the multiple stages skipped in hepatocarcinogenesis. Unlike for the other cancers, therefore, infection with HCV may be capable of eliciting HCC in the absence of a complete set of genetic aberrations. Such a scenario would explain unusually high incidence and multicentric nature of HCC developing in chronic hepatitis C. PMID- 11494536 TI - [Effects of hepatitis C virus proteins on the interferon-stimulated signal transduction]. AB - The mutation number in the interferon sensitivity determining region(ISDR) (aa2209-2248) in the nonstructural 5A(NS5A) region of hepatitis C virus(HCV) has been shown to correlate with the sensitivity of the virus to interferon treatment. To clearify this mechanism, the functions of the NS5A protein have extensively been studied. The NS5A protein has been shown to be phosphorylated. The NS5A protein 1) is a transactivator, 2) represses the expression of interferon-stimulated genes, 3) binds with PKR, an interferon-stimulated gene, and inhibits its activity, and 4) may inhibit interferon-induced apoptosis of HCV infected hepatocytes. These effects of the NS5A are different depending on the ISDR types. The E2 protein also can bind with PKR and inhibits its activity. These mechanisms may explain the low responsiveness of HCV to interferon treatment and the difference in the sensitivity to interferon among the genotypes and ISDR types. PMID- 11494538 TI - [Interferon-treated hepatitis C virus(HCV) patients with sustained biochemical response without eradication of HCV(asymptomatic HCV carrier)]. AB - Patients with hepatitis C virus(HCV) responding differently to interferon(IFN) therapy were speculated to have different incidence of disease progression to cirrhosis and of the development of hepatocellular carcinoma(HCC). However, the background and prognosis of the patients with sustained biochemical response without eradication of HCV (BR) (asymptomatic HCV carrier) has not been revealed so far. Review of recent studies suggest that the characteristics of the patients with BR are lower HCV RNA load, higher rate of HCV subtype-2 and lower score of liver fibrosis when compared with those with NR. The IFN therapy in patients who have not cleared HCV and showed normal ALT retards progression of fibrosis and reduces the incidence of cirrhosis and HCC. PMID- 11494537 TI - [Treatment of chronic hepatitis C patients complicated with the other disease]. AB - Because of relatively low efficacy, considerable side effect and high cost, HCV infected patients complicated with the other disease(patients with hematologic dysorders, chronic renal failure on hemodialysis, HIV co-infection and collagen disease) have been excluded from large trials evaluating the efficacy of IFN or in combination with ribabirin. So, little is known about treatment of these patients. We analyzed the medical literature focusing on treatment of HCV infection in these patients, to suggest conclusion about indication based on tolerance and efficacy. In case the other disease is well controlled and long term sevival is expected, treatment should be required based on the evaluation of liver function test including liver pathology for preventing liver disease from developing terminal stage. Further studies are needed to better define HCV therapies in these patients. PMID- 11494539 TI - [Hepatitis C in hemodialysis patients]. AB - Characteristics of chronic hepatitis C in 99 hemodialysis patients were studied. Eleven out of 18 anti-HCV-positive patients(61.1%) and 13 out of 81 anti-HCV negative patients (16.0%) were HCV-RNA-positive. Twenty four HCV-RNA-positive patients were divided into 2 groups according to anti-HCV-positivity. Seven out of 11 patients in anti-HCV-positive group and 1 out of 13 in anti-HCV-negative group were infected with more than 100 K copies/ml of HCV. A number of HCV-RNA copy was significantly larger in anti-HCV-positive group. Fourteen biopsies and necropsies in hemodialysis were achieved and the histological diagnosis did not correspond to clinical feature of chronic hepatitis C. PMID- 11494540 TI - [Hepatitis virus C infection in children]. AB - The prevalence of hepatitis C virus(HCV) infection has been becoming much lower in childhood. Blood transfusion was the principal transmission route of HCV in children. Since the year of 1989, screening of the blood products for HCV antibodies has markedly reduced transfusion-related HCV infection. Mother-to infant transmission occurs in less than 10% of infants born to HCV-infected mothers. The maternal viral load is the most important factor determining the risk. The natural history and the outcome of HCV infection in childhood dependent on host and viral factors. The rate of progression to chronicity is 60% in post transfusion infection and 80% in vertical infection. Experience of interferon therapy of chronic hepatitis C in children is limited, with about 50% having a sustained response to the therapy. PMID- 11494541 TI - [Nosocomial infection with hepatitis C virus(HCV)]. AB - Nosocomial infection with HCV occurs in three ways. 1) Infection from medical personnel to patients is extremely rare, and only one report exists. Medical personnel should be aware of HCV infection status, but HCV-infected medical personnel need not be banned from medical activity. 2) The possibility of infection from patient to medical personnel is always present. The creation of a medical environment in which the possibility of injury from instruments contaminated by blood is eliminated is the most important measure. The probability of infection is estimated to be around 1%, and there are no methods of prevention. Interferon therapy is administered when acute hepatitis C infection develops. 3) Patient to patient infection is thought to have occurred frequently in the past as a result of re-use of syringes for intravenous injections. The most important measure against this occurrence is the strict disposal of disposable syringes. PMID- 11494542 TI - [The present state and problems of HCV blood screening system in blood products]. AB - After discovering HCV antibody in 1989, the new cases of posttransfusion hepatitis C has been rare. This HCV antibody screening system provided certainly the powerful preventive effect from HCV infection through blood products. But it was possible to infect HCV through the blood products without HCV antibody but with HCV-RNA. This phenomenon seemed to be observed in a window period between time at infection and antibody appearance. To detect the infectious source at a window period in blood products, NAT(nucleic acid amplification) method was adapted recently in Japanese red cross. For safe blood screening system, continuous efforts such as NAT will be needed forever. PMID- 11494543 TI - [A trial of new interferon therapy for the patients with chronic hepatitis C resistant to interferon therapy]. AB - HCV-RNA clearance rates are reported to be about 30-40% of the patients treated with IFN for less than 6 months. But IFN therapy is ineffective for chronic hepatitis C patients with HCV-genotype 1b and a high virus load. We evaluated the efficacy of different IFN therapy compared with standard IFN therapy for chronic hepatitis C patients resistant to interferon therapy. That is, we assessed the following therapy; 1) prolonged IFN therapy, 2) IFN-beta therapy of twice a day, 3) IFN-beta therapy daily for 24 weeks, 4) combination therapy of IFN-alpha and IFN-beta. In some cases, these IFN therapies were effective than a standard IFN therapy. PMID- 11494544 TI - [Two-step interferon rebound therapy for refractory chronic hepatitis C]. AB - In almost all patients in whom interferon(IFN) treatment dose not result in persistent negative findings for HCV-RNA, HCV-RNA levels show a rebound after discontinuing the administration. In most patients, HCV-RNA levels after administration are increased compared to pretreatment values. When a rapid increase in HCV-RNA levels causes transient exacerbation of transaminase levels, HCV-RNA levels then rapidly decrease. In two-step interferon rebound therapy(TIRT), IFN is additionally administered when HCV-RNA levels are decreased. We previously reported that TIRT was useful for treating type 1b patients with an HCV-RNA level of 1 Meq/ml or more who did not respond to IFN treatment. In the year 2000, health insurance covered the additional administration of IFN. The use of TIRT for additional administration may further improve treatment response in patients who do not respond to IFN treatment. PMID- 11494545 TI - [Combination therapy of IFN and ribavirin]. AB - Ribavirin is a purine nucleoside analog that inhibits the replication of a wide range of DNA and RNA viruses, although ribavirin treatment dose not significantly decrease serum virus load in patients with chronic hepatitis C. Several lines of evidence from the randomized large-scale studies of USA and Europe indicate that combination therapy with ribavirin significantly enhances the sustained response rate of IFN therapy in major patient types with chronic hepatitis C. A recent trial in Japan also proved that the combination therapy is superior to IFN monotherapy. Thus, IFN-ribavirin combination is likely to become the antiviral therapy of choice for chronic hepatitis C. PMID- 11494546 TI - [IFN combined cyclosporin A therapy]. AB - Current therapy of chronic hepatitis C with a 6 months course of alpha interferon shows limited effectiveness for patients with chronic hepatitis C. In particular more effective therapy is needed to treat patients who have high serum level of group 1-HCV RNA. We previously reported that a combination of interferon and cyclosporin A was more effective than interferon monotherapy. We also examined whether CsA has an antiviral effect on HCV using cultured human hepatocyte cell line. Propagation of HCV seems to be inhibited by CsA in a dose dependent manner. Further study is needed to clarify the action of cyclosporin A. PMID- 11494547 TI - [Interferon therapy to chronic hepatitis type C for the prevention of hepatocarcinogenesis]. AB - Interferon(IFN) therapy for chronic hepatitis(CH) related by hepatitis C virus is useful for the prevention of the appearance of hepatocellular carcinoma(HCC) by both prospective and retrospective study. IFN could be reduced an activity of necro-inflammatory reaction leading toward the reduction of fibrogenesis. Therefore, IFN treated group had a low potential carcinogenesis of the liver indicating the prevention of HCC from CH type C, even if virological complete remmision(CR) could not be obtained after IFN treatment. Biochemical response(BR) group as well as CR group could be inhibited hepatocarcinogenesis compare with non-IFN treated group. Recently, IFN applied for liver cirrhosis as same concept for the prevention of HCC. PMID- 11494548 TI - [Hepatocellular carcinogenesis and prognosis of elderly patients with chronic hepatitis type C]. AB - Among 457 elderly patients of 65 years or older with chronic hepatitis or cirrhosis caused by hepatitis C virus, 117 patients underwent interferon therapy for the elimination of hepatitis C virus. A total of 87 patients could be analyzed for the interferon effect, since the remaining 20 patients had still been receiving or just finished the therapy. Thirty-six patients(41.4%) achieved complete elimination of HCV-RNA with interferon therapy. Although those patients with a milder hepatitis stage and better virological condition(low viral concentration or group 2 subtype) were preferentially enrolled in the therapy, 13 patients(11.1%) discontinued the administration with varied side effects: severe general malaise in 6 patients, depression in 3, pneumonia/pneumonitis in 2, and retinopathy in 2. Crude hepatocellular carcinogenesis rates in the subgroup of F1 + F2 and the subgroup of F3 + F4 were 1.8%, 21.2% at the end of 5th year, and 14.3% and 53.7% at the tenth year, respectively. PMID- 11494549 TI - [A possible role of human leukocyte antigen(HLA) typing for predicting response to interferon therapy in chronic hepatitis C]. AB - Hepatitis C viral load, genotype and/or staging of liver fibrosis are known to be factors for predicting response to interferon(IFN) therapy in patients with chronic hepatitis C. The aim of this study is to investigate if human leukocyte antigen(HLA) typing is related to the response to IFN therapy. The seventy six Japanese patients were studied and categorized into two groups: 46 patients with chronic hepatitis C (Group A) and 30 with liver disease unrelated to HCV infection(Group B). In addition, 39 patients who were treated with IFN were classified into complete responders(CR) and non-complete responders (NR). There was not any differences in HLA typing between group A and B, but the frequency of HLA class I B51(5) was higher in CR than in NR patients(p = 0.045). When restricted to those who had low viral load(under 10(55) copies/ml) and genotype 2a or 2b, HLA class I CW1 was found in 7 responders(70%) and in 1 non responder(14%) (p = 0.023). HLA class II DR9 was not found in responders but in 3 non-responders(p = 0.022). These preliminary results suggest that HLA types may be related response to IFN therapy in patients with chronic hepatitis C. PMID- 11494550 TI - [Type I IFN receptor]. AB - Interferon(IFN) alpha and IFN beta apparently share the same receptor structure, the type I IFN receptor, which consist of at least two subunits. The subunits are referred to as IFN AR1(IFN alpha receptor) and IFNAR2(IFN alpha/beta receptor). As IFN elicits antiviral activity by binding to receptors on the cell surface, the expression of the type I IFN receptor in liver may be directly involved in the pathogenesis of viral hepatitis and response to IFN therapy. Here we discuss if the IFN receptor expression in liver and peripheral blood mononuclear cells can be a predictive factor for the response to IFN therapy in patients with chronic hepatitis C. PMID- 11494551 TI - [HCV genotype as a predictor of response to interferon therapy in patients with chronic hepatitis C]. AB - Hepatitis C virus(HCV) genotype is one of the most important predicting factors of response to interferon(IFN) therapy in patients with chronic hepatitis C. According to the molecular evolutionary analysis, HCV is classified into six major genotypes. The patients infected with genotype 1 show high HCV RNA levels and poor response to IFN therapy compared to those with genotype 2 or 3. No sufficient data are observed on response to IFN in patients with genotype 4 to 6. When PEG-IFN plus ribavirin therapy is introduced, high proportion of patients without genotype 1 must show complete response. In the near future, to predict good response to IFN therapy, it will be necessary to know whether patients have HCV genotype 1 or not. PMID- 11494552 TI - [Dynamics of serum HCV RNA levels during IFN therapy in patients with chronic hepatitis C for prediction of outcome of IFN therapy and beneficial dosing]. AB - For beneficial Interferon therapy to patients with chronic hepatitis C, we assess the change of serum HCV RNA levels during the early phase of IFN therapy, comparing the schedule of IFN beta 3 MU x 2 times per day and daily administration of IFN alpha 2b 10 MU once per day. During first 2 weeks, decrease in serum HCV RNA levels was significantly faster in IFN beta 2 times per day than alpha 2b. We concluded that the administration of IFN beta 2 times per day for 2 weeks followed by IFN alpha 10 MU will be suitable for IFN induction therapy. PMID- 11494553 TI - [Present status of clinical trials on long-acting IFNs preparations (PEG-IFN alpha 2a, alpha 2b, IFN alpha-minipellet)]. AB - Interferon(IFN) is an essential component of the treatment of chronic HCV infection and at present, it is the most important to improve its efficacy, not only for HCV chronic liver diseases, but also for the prevention of HCV associated hepatocellular carcinoma. Two long-acting IFN preparations(PEG-IFN alpha 2a and 2b) have been used at present and clinical studies have shown that sustained virologic, biochemical and histological responder rates are significantly higher in PEG-IFN-treated patients with HCV associated chronic hepatitis and cirrhosis comparing with ones treated with conventional IFN. In addition, PEG-IFN treatment in combination with ribavirin seems to be the best for HCV-associated chronic liver diseases. PMID- 11494554 TI - [Immunoadsorption therapy for HCV infected chimpanzee]. AB - Response of Interferon therapy to chronic hepatitis C patients are poor who infected high titer of hepatitis C virus(HCV) RNA. It has been reported that HCV particles coexist low density lipoprotein(LDL) or anti-HCV antibodies as antigen antibody complex in chronic hepatitis C patients. We demonstrated the ability of removal HCV particles by these adsorbents and the efficacy of HCV RNA reduction by immunoadsorption therapy for HCV infected chimpanzee. By batch method, IgG and LDL adsorbent showed 66% HCV RNA reduction and IgG adsorbents kept better adsorption ability than that of others when adsorbent: serum ratio was increased. Immunoadsorption therapy to chronic HCV infected chimpanzee was carried out on line with a plasma separator. HCV RNA titer of this chimpanzee decreased 35% immediately after therapy, but after 4.5 hours, it recovered as same level as that of before procedure. PMID- 11494555 TI - [The present studies of the development of HCV vaccine]. AB - After isolation of a hepatitis C virus genome in 1989, all donated blood samples have been analyzed by a sensitive screening system, which brought us 'safety transfusion' (free from HCV). However it is likely that about 170 million people around the world, more than 2 million people in Japan have already infected with HCV. Despite the numerous efforts, the lack of efficient cell culture system makes it difficult to develop HCV vaccine. Some novel strategies are engineered day by day that might be useful tools. Hereafter we must clarify the mechanisms of replication and infection in depth, to develop a vaccine that clear HCV from carrier. PMID- 11494556 TI - [Epidemiological trend of chronic hepatitis C]. AB - Though patients with chronic hepatitis C after blood transfusion hepatitis decrease dramatically during the past decade, patients with it after non transfused acute hepatitis C are still in existence. This means that new patients with chronic hepatitis C are on the decrease but not diminish. To the contrary, the number of patients with chronic hepatitis C is increasing. This depends on mainly dig up the undiscovered patients. These result in increasing the patients with hepatocellular carcinoma. PMID- 11494557 TI - [Current status of tissue engineering for therapeutic use]. AB - During the past three decades, significant advances have been made in the field of transplantation. However, clinical obstacles such as donor scarcity, cost, infection and life-long immunosuppression(especially within the pediatric age group) have become the major limitations for organ transplantation. This article discusses the recent progresses as well as the remaining challenges in tissue engineering that may enable such technology to replace organ transplantation. Additionally, we describe a practical method for tissue engineering in the field of cardiovascular surgery. PMID- 11494559 TI - [Progress in the research of intractable pancreatic diseases in Japan]. PMID- 11494558 TI - [Ghrelin: a novel growth-hormone releasing peptide]. AB - Growth hormone secretagogues are small synthetic molecules that act through an orphan G-protein coupled receptor(GPCR) called GHS-R. We purified its endogenous ligand from rat stomach and named it 'ghrelin'. Ghrelin is a 28-amino-acid peptide with an essential n-octanoyl modification at Ser3. The discovery of ghrelin indicates that the release of GH from the pituitary may be regulated not only by hypothalamic GHRH, but also by ghrelin from the stomach and hypothalamus. Moreover, ghrelin induces appetite-stimulating effect and adiposity in rat. These facts indicate that ghrelin is a natural leptin antagonist. PMID- 11494560 TI - [Current topics on classification and nomenclature of bacteria. 6. Classification of Mycobacteria and future perspectives]. PMID- 11494561 TI - [Costs of influenza therapy]. AB - The costs of outpatients with influenza. Outbreak of influenza occurs every winter in Japan and it brings a huge impacts on society and the individual. We calculated the costs of medications and medical examinations for outpatients with influenza in hospital. After we confirmed the diagnosis of influenza by rapid diagnosis system (Directigen Flu A) or hemagglutinin inhibition test, neutralization test, comparison of costs was done between influenza patients (89 cases) and non influenza patients (212 cases). Mean cost of total medication and medical examinations were 14,800 +/- 980 yen for an influenza patient and 12,420 +/- 976 yen for a non influenza patient. This data showed that treatment of the influenza patient is more expensive than non influenza patient. It must be considered that prophylaxis with influenza vaccine and treatment with antiinfluenza drugs given in the early stage of influenza are useful for reducing medical costs. PMID- 11494562 TI - [Clinical and bacteriological features of 12 cases of liver abscess caused by Streptococcus milleri group]. AB - We described the clinical and bacteriological features of 12 cases of liver abscess caused by Streptococcus milleri group (SMG) during a 6-year period from 1993 to 1998. The gender was 11 males and 1 female with their ages ranging from 39 to 76 years old (mean: 53.4). The common symptoms were fever (100%), abdominal pain (67%), and appetite loss (58%). Nine cases had underlying diseases such as carcinomas and diabetes mellitus. Predominant causes of the liver abscess were cryptogenic (42%) and biliary tract disease (33%). Three patients died of an exacerbation of the carcinoma. Eight cases (67%) was single infection of SMG and no mixed infection with anaerobes. No strains isolated in this series showed resistance against penicillin G and ampicillin. SMG was highly isolated from the blood culture in eight of the 11 cases (73%). Liver abscess should be taken into consideration as one of the causes of SMG septicemia. PMID- 11494563 TI - [Five cases of erythema infectiosum in adults]. AB - The similarities between clinical features of erythema infectiosum and collagen disease or other viral infections prompted us to investigate clinical manifestations and laboratory data of parvovirus B19 (B19) infection in adults. We diagnosed all five patients as acute B19 infection by antibody assays. The age of patients ranged from 18 to 39 years old (mean 29), and all patients were female. All five patients showed high fever, arthralgia and edema of the extremities. Four of the five patients showed skin rash of the extremities or cheeks. Two patients were diagnosed as erythema infectiosum by family physicians before coming to us. The three remaining patients were suspected to be systemic lupus erythematosus, adult Still disease or rubella indivisually and referred to our hospital. A-27-old female (case 5) visited our hospital because of polyarthralgia and butterfly rash on her face. A test for antinuclear antibodies (ANA) was positive at a dilution of 1:320. Rheumatoid factor (RF) was also detected by latex fixation test. Her AST was 51 IU/L, ALT 68 IU/L and LDH 568 IU/L. Her symptoms persisted for 3 weeks and hepatic dysfunction recovered within 3 weeks. Five months later. ANA was negative at the dilution of less than 1:40. We suggest that the similarities between some symptoms of B19 infection and clinical and serological manifestation of collagen diseases merit closer attention. PMID- 11494564 TI - [Investigation of infective endocarditis clinical isolates of methicillin resistant Staphylococcus aureus non-responsive to vancomycin]. AB - Methicillin resistant Staphylococcus aureus (MRSA) is one of the most important strains which induce hospital and post-operative infection. In cases of infective endocarditis in which VCM was not efficacious, MRSA strains were chronologically isolated at three different times and examined with the following parameters: minimum inhibitory concentration (MIC), fractional inhibitory concentration (FIC) index, Mu 3 agar, population analysis, pulse field gel electropholesis (PFGE). The PFGE banding patterns of the three MRSA isolates were the same, therefore, it was concluded that the same strain of MRSA was selected for reduced susceptibility. A pattern of Mu 3 and Mu 50 was demonstrated under population analysis. PMID- 11494565 TI - [Outbreak of measles in a hospital and measures taken against hospital infection- evidence of cost and benefits]. AB - In Japan, an isolated vaccine of measles is used because MMR vaccines have been suspended due to the frequent occurrence of aseptic meningitis after their use. It is administered only once with the cover rate having been approximately 70%. An outbreak of measles was experienced in eight of our health care workers (three doctors, three nurses and two clerks of our hospital) and in seven of our medical students, accompanying local outbreaks. Their condition was severe enough to require admission to our school hospital. One of medical students developed encephalitis, but he has recovered completely. The social cost including medical cost per worker was approximately yen 500,000 (about $4,500). We measured antibodies against measles, rubella, chickenpox and mumps using the ELISA assay in 1048 health care workers less than 40 years of age and 99 medical students before their clinical practice two or three months after the outbreak. The cost including the measuring of antibodies and vaccination for these workers and students was approximately yen 2,800,000 and it will be yen 700,000 for new workers each year. The negative rates for workers were 1.2% for measles, 8.8% for rubella, 10.0% for mumps and 2.0% for chickenpox and those for the medical students were 1.0%, 22.0%, 9.0% and 5.0%, respectively. Of the workers, 9.6% who had more than the defined very high tiaer were thought to have subclinical reinfection because they were non-symptomatic. The rates were 1.2% for rubella, 0.5% for chickenpox and 0.0% for mumps for more than the defined high titers among the workers, suggesting a risk of outbreaks in the future. Therefore, the antibodies of health care workers and medical students should be measured at the hospital and the antibody-negative person should be vaccinated to prevent infection not only from their patients but also prevent other patients from becoming infected by contracted health care workers or students. PMID- 11494566 TI - [The trends of Vibrio parahaemolyticus foodborne outbreaks in Tokyo: 1989-2000]. AB - The total number of foodborne outbreaks due to Vibrio parahaemolyticus in Tokyo during the last 12 years between 1989 and 2000 were 710. The number of outbreaks in a year was 55 in 1989, 75 in 1990, and there was a gradual decrease to 24 outbreaks in 1993 which was the smallest number during those 12 years. After 1994, the number of outbreaks increased dramatically year by year until 1998 (107 outbreaks). Then they had decreased slightly to 74 in 1999, 65 in 2000. The monthly incidence of V. parahaemolyticus foodborne outbreaks showed a peak in August (44.2%) each year. In the last 12 years, 88.7% of V. parahaemolyticus foodborne outbreaks occurred during the 3 months between July and September, while 99.9% occurred between June and October. The most prevalent serotype of V. parahaemolyticus also changed, the most prevalent was O4:K4 in 1989, O4:K8 in both 1990 and 1991, O1:K56 in 1992, and O4:K8 from 1993 through 1995. Serotype O3:K6 became the most prevalent in 1996 and has remained so to date. In addition, the new serotype O4:K68 had also appeared in 1998. The number of outbreaks due to serotype O4:K68 followed that of O3:K6. Thus, the trends of V. parahaemolyticus foodborne outbreaks during the last 12 years in Tokyo showed various characteristics and dramatic changes in causal organisms. PMID- 11494567 TI - [Characterization of enterovirus type 71 isolated in Saitama Prefecture in 2000]. AB - Coxsackievirus A16 (CA16) has been isolated mainly from hand, foot and mouth disease patients for the past several years in Saitama Prefecture, Japan, but in 2000, enterovirus 71 (EV71) was isolated with high frequency. Also, many cases with aseptic meningitis were reported unusually. We determined partial nucleotide sequences of EV71 isolated in this year and compared them with those of the viruses isolated previously. Eighteen strains of EV71 were isolated from May to September in 2000. The sequence analysis of about 700 nucleotides between 5' NCR and VP2 region indicated that the EV71 strains isolated in Saitama Pref. in 2000 were classified into three groups. The first group was consisted of the strains which showed 89 to 91% nucleotide similarity with TW/2086/98, NCKU 9822 isolated in Taiwan and strain SA/448/99 isolated in Saitama Pref. in 1999. The second group was highly similar to both Taiwan strains and SA/448/99 (92 to 93% nucleotide similarity). The first and second groups were also considered to be subgroups belonging to the same group. And the third group was highly similar to 18/Sin/97, 13/Sin/98 isolated in Singapore. These viruses were genetically close to MS7423/87, which has shown neurovirulence. Furthermore, from geographical origin of specimens, we found that the Taiwan like strains were isolated in the west area (Kawagoe, Tokorozawa) and Singapore like strains were isolated in the south area (Urawa, Kawaguchi, Toda). Also, almost every EV71 strains isolated from the patient with aseptic meningitis were close to Singapore strains. It was found that EV71 strains isolated in 2000 were different from the strains isolated past in Saitama Pref. and some of them were close to the neurovirulent strain. PMID- 11494568 TI - [A case presenting severe respiratory failure with high antibody titers to Chlamydia pneumoniae]. AB - "HITAZYME C. pneumoniae" is a diagnostic reagent that has been recently developed by adopting an ELISA method for detection of anti-Chlamydia pneumoniae (C. pneumoniae) antibodies. A case presenting bilateral interstitial opacities and severe respiratory failure with high titers of "HITAZYME C. pneumoniae" was described. Sputum, blood, serological, and bronchoalveolar lavage examinations failed to reveal other etiology to explain his severe respiratory illness. Clinicians should be aware that C. pneumoniae may cause severe respiratory failure or ARDS. Further studies are needed to evaluate the role of C. pneumoniae infection in the development of severe pneumonia or ARDS. PMID- 11494569 TI - [An adult case of systemic cat-scratch disease with hepatosplenic involvement]. AB - A previously healthy 25-year-old female was admitted to our hospital in November, 1997, for treatment of a spike-fever of 2 weeks' duration. She had a cat in her house but reported no history of cat bites or scratches. No peripheral lymphadenopathy was detected. White blood cell count was within normal limits, but an increased C-reactive protein level of 11.4 mg/dl was noted. Infectious disease was suspected but ruled out as blood cultures were negative. Empiric therapy with clarithromyoin, isoniazid, and rifampicin was ineffective. In January, 1998, abdominal ultrasonogram revealed multiple hypoechoic mass lesions in the spleen and liver, and a splenectomy was performed in March. Histopathologic examination showed numerous necrotizing and caseating granulomas, which tested positive for Bartonella henselae DNA by PCR. Furthermore, the patient tested positive for B. henselae antibody by immunofluorescence assay. A diagnosis of systemic cat-scratch disease with hepatospnenic involvement was made. Combination therapy with minocycline, sulbactam/cefoperazone, and tosufloxacin was administered and her inflammatory findings improved gradually. We report an adult case of systemic cat-scratch disease with liver and spleen involvement in the non-immunocompromised host. PMID- 11494570 TI - [A case of Mycobacterium avium lung infection with a pleural effusion]. AB - Pleural effusions seldom accompany nontuberculous mycobacterial infections. We reported one such case of M. avium lung infection with pleural effusion. A 40 year old male was admitted to our hospital complaining of right chest pain and general fatigue. His chest X-ray showed a consolidation in the right lower lung field. The day after admission, a right pleural effusion appeared. The fluid was exudative and microbiological examinations of the effusions, including staining and culturing, proved negative. However, one month afteradmission, acid fast bacilli were observed in his sputum and a subsequent sputum culture specimen revealed the presence of M. avium. Treatment with antimycobacterial agents was promptly commenced and the patient's effusion and lung consolidation was gradually resolved. PMID- 11494571 TI - [Development of gait in childhood: a systematic gait analysis]. AB - To study the normal gait development in childhood, we performed a systematic gait analysis on 37 normal children (20 boys and 17 girls) aged between two and six years. The gait cycle, single support phase, ratio of stride length to body height, plantiflexor angles of the feet and propelling force increased with age, whereas the ratio of gait width to stride length and dorsiflexor angles of the feet decreased. The knee-flexion waves, heel-strikes were found in all subjects. From two to six years of age, balance-holding ability and feet plantiflexor forces developed, resulting in more efficient locomotion. The pattern of the lower limb movements on gait is considered to mature during the first three years of life. PMID- 11494572 TI - [Medical and educational support to children with neuromuscular disorders in schools: an analysis based on a questionnaire to teachers]. AB - To clarify the medical and educational support necessary for ordinary school life of children with neuromuscular disorders, a questionnaire was sent to their homeroom teachers. The 13 outpatients consisted of 7 with Duchenne muscular dystrophy, 5 other muscular dystrophies, and 1 with another myopathy. Their teachers desired the environment and system, such as elevators and personnel to support their movements, should be improved. Regarding the school curriculum, special advice of medical staff was considered to be necessary to provide physical education and to promote their well-retained activities. The teachers regarded it valuable to teach them together with healthy children, for the children with or without neuromuscular disorders, teachers, and people in the community. In the future, it is expected that national sanatariums and schools for the handicapped will function as special facilities that are open to the community for general counselling and provide various informations. Pediatric neurologists should play a role as managers caring for the patients, their families, teachers, schools, and communities. PMID- 11494573 TI - [Clinical evaluation of treatment with fluvoxamine, a selective serotonin reuptake inhibitor in children with autistic disorder]. AB - The serotonin system has been implicated in the pathoetiology of autistic disorder. To examine the clinical effects of fluvoxamine, a selective serotonin reuptake inhibitor (SSRI) in children with autistic disorder, eighteen patients underwent a cross-over, double-blind trial of fluvoxamine treatment after a written informed consent was obtained from patients' parents. Fluvoxamine treatment resulted in significant improvements in some clinical findings such as eye contact and language use, as tested by behavioral assessment scores consisting of twenty items (p < 0.05). The improvement in language use was also confirmed by parental assessments. Clinical Global Impression Scale was improved in approximately half of the patients. No severe adverse effect was observed during the trial. Thus SSRI treatment in autistic children may be of value. PMID- 11494574 TI - [Clinical and MRI findings in childhood idiopathic intracranial hypertension]. AB - We report here clinical and MRI findings of four children with idiopathic intracranial hypertension (IIH). Their chief complaint was headache. Three patients had papilloedema, and the other one showed the highest cerebrospinal fluid pressure (106 cm H2O) without papilloedema. In two cases, the symptoms disappeared after repeated lumbar punctures. At admission, MRI demonstrated empty sella in all the four cases. Regarding the optic nerves, there were more than two of the following three findings: distension of the perioptic subarachnoid space, vertical tortuosity and elongation of the optic nerve, and flattening of the posterior aspect of the globe. Follow-up MRI showed normalization of sella turtica and/or optic nerve findings in two of the three cases examined. MRI focusing on the optic nerves and pituitary gland may provide important clues for the diagnosis of IIH, especially those without papilloedema. PMID- 11494575 TI - [Methylphenidate therapy in 141 patients with hyperkinetic disorder or with pervasive developmental disorder and hyperkinesia]. AB - The efficacy and adverse reactions produced by methylphenidate (MPD) therapy were evaluated in 141 patients with hyperkinetic disorder or pervasive developmental disorder (PDD) with hyperkinesia. Ninety-nine patients were followed for 1 to 5 years to determine if the treatment could be continued and if the patients' adaptation to their environment improved. The results showed that the MPD therapy was effective in 93% of patients whose IQ was > 80 and in 70% whose IQ was < or = 80. The efficacy was not significantly different between patients with PDD and those without PDD. Of the patients in whom the MPD therapy was effective, the majority received a MPD dosage of 0.3 mg/kg once every morning. Adverse reactions, such as excitability, nausea or anorexia, and insomnia were reported in 23% of the patients. Although this figure was not negligible, no serious events occurred. Seizure induction was suspected in 2 patients. Many of the patients (53/83) in whom the MPD treatment was effective continued to receive the treatment throughout the follow-up period. By the time that the conditions were alleviated to the extent that the treatment could be stopped, the patients had become well adapted to their environment. However, in many other cases, adaptation was unsatisfactory. In these cases, psycosocial interventions were necessary, even if the MPD therapy was effective. PMID- 11494576 TI - [Proton magnetic resonance spectroscopy of the autistic brain]. AB - To evaluate brain dysfunction in autism, proton magnetic resonance spectroscopy (1H-MRS) was performed for 29 autistic patients (5-15 y.o.) and 19 normal children (6-14 y.o.). We obtained magnetic resonance (MR) spectra of the left and right amygdaloid-hippocampal regions and the left cerebellar hemisphere with a STEAM sequence (TR = 5000 ms, TE = 18 ms). In addition to the evaluation of signal intensity ratios, the absolute concentration of three major metabolites (N acetylaspartate [NAA], creatine/phosphocreatine [Cr] and choline-containing substances [Cho]) was quantified by an internal reference method using unsuppressed tissue water. Although no abnormal MR images were found in the three regions examined, the signal intensity and the concentration of NAA in the left amygdaloid-hippocampal region and the left cerebellar hemisphere were reduced significantly in autistic patients compared to normal children. We speculated that this decrease in NAA reflected neuronal loss, immaturity or hypofunction in these regions. The results of our study were in agreement with those of previous studies on autism, one by neuropathological methods and the other using a single photon emission computed tomography with 99mTc HMPAO. Disorders of the amygdaloid hippocampal region and cerebellum are considered to play an important role in the characteristic cognitive and emotional dysfunction in autism. 1H-MRS is a valuable tool to clarify the pathophysiology of autism. PMID- 11494577 TI - [Recurrence of febrile convulsions after the first diphtheria-pertussis-tetanus vaccination and measles vaccination in children with febrile convulsions: a questionnaire survey in Takamatsu City]. AB - A questionnaire about convulsions and other adverse events after vaccination was sent to doctors who administered a diphtheria-pertussis-tetanus (DPT) vaccine (the first dose) or a measles vaccine between April 1, 1995 and December 31, 1997 in Takamatsu City to children with convulsions. DPT and measles vaccines were administered to 300 and 339 such children, respectively. Many of them had febrile seizures, the last of which had occurred before more than 1 year. Among them, 175 cases were administered with DPT and 180 with measles vaccine. There were recurrences of febrile convulsions after immunization in 2 (1.1%) of the cases given DPT and 3 (1.7%) of those given measles vaccination. According to the data of the Monitoring System for Adverse Events Following Immunization (the Ministry of Health and Welfare of Japan), the incidence of convulsion after immunization in healthy children between April 1, 1996 and September 30, 1997 was 0.4% after the first dose of DPT vaccination and 0.3% after measles vaccination. In comparison, the incidence was higher in children who had had febrile convulsions before more than one year. Especially, the rate of convulsions after measles vaccinations was significantly higher (p < 0.05) in children with febrile convulsions. These results suggest that the measles vaccination should be administered with caution to the children with previous febrile convulsions. PMID- 11494578 TI - [Mesial temporal lobe epilepsy in a patient with Wilson's disease receiving FK506 (tacrolimus) after liver transplantation]. AB - Mesial temporal lobe epilepsy (MTLE) developed in a boy receiving FK506 (tacrolimus) after liver transplantation. He had no history of convulsions. At the age of 7, he underwent liver transplantation 13 days after he developed the abdominal form (fulminant hepatitis) of Wilson's disease. On postoperative day 18, he had a generalized tonic seizure (duration 20 min.) with loss of consciousness. FK506 was discontinued under the suspicion of FK506-induced encephalopathy. His symptoms resolved within a few days. FK506 was readministered at 3 months after transplantation. Ten months later, he developed complex partial seizures characterized by right tonic posturing with oral automatism. EEG revealed sporadic spikes in the anterior temporal region. MRI and SPECT showed bilateral (left side dominant) hippocampal lesion, which suggested the diagnosis of MTLE. Since seizures became refractory to medical treatment with progressive worsening of memory functions, FK506 was discontinued again at 36 months after readministration. Six months later, his memory improved remarkably, but there were no changes in seizure frequency and in MRI and SPECT findings. Our findings indicate that FK506 might damage the hippocampus, thereby causing MTLE. Additional case reports, however, will be required to elucidate this new FK506 related neurological complication. PMID- 11494579 TI - [Fatal stenosis of trachea and main bronchus due to compression by the spine and sternum in three patients with severe motor and intellectual disabilities]. AB - Three patients with severe motor and intellectual disabilities developed fatal respiratory insufficiencies caused by stenosis of the trachea and main bronchus due to compression by the spine and trachea. The onset of respiratory insufficiency was by 20, 16, and 29 years of age. Chest computed tomography demonstrated deformation and narrowing of the trachea and main bronchus, and shortening of the distance between spine and sternum. Although respiratory insufficiency was temporarily relieved by insertion of a stent into the main bronchus in patient 1, he died at the age of 21 due to proliferation of granulation tissue. Patient 2 died of bleeding from the granulation tissue around the window of tracheotomy at the age of 18 years, and patient 3 died of recurrent pneumonia at the age of 34 years. In conclusion, stenosis of the trachea and bronchus observed in these patients was caused by progressive deformation of the thorax. The stenosis may result in sudden death or recurrent respiratory infections in patients with severe motor and intellectual disabilities. PMID- 11494580 TI - [A case of peculiar type of post-encephalitic/encephalopathic epilepsy: efficacy of bromide in the control of intractable seizures]. AB - A 5-year-old boy with a peculiar type of post-encephalitic/encephalopathic epilepsy is reported. He had been healthy showing normal development before its onset. Five days after the onset of an upper respiratory infection, he had a severe generalized seizure, that evolved into intractable seizures. They were highly resistant to almost all anticonvulsants and occasionally resulted in status epilepticus. High-dose phenobarbital therapy successfully controlled the convulsions, but was discontinued because of drug-induced aplastic anemia. Alternative bromide therapy was markedly effective in controlling the seizures. PMID- 11494581 TI - [A girl with chief compliance of "easy to forget"--cognitive neuropsychological evaluations and cerebral blood flow]. AB - Cognitive-neuropsychological evaluations were performed on a 12-year-old girl with memory disorder to study the higher brain function of this patient. The cerebral blood flow (r-CBF) was also studied. The results showed that the auditory-verbal memory of this patient were impaired. The regional cerebral blood flow was reduced in the left hippocampus which reportedly cause memory disorder. The disturbance of visual cognition and disability of construction in this patient seem to be related to reduced cerebral blood flow in the left parietal region. PMID- 11494582 TI - [Nucleotide sequence analysis of the hepatitis B virus X-precore region in chronic hepatitis B]. AB - In chronic hepatitis B virus (HBV) infection, hepatitis B e antigen (HBeAg) is seroconverted from positive to negative. HBeAg is associated with active replication of the HBV and a high disease activity. To elucidate the relation between HBV X-precore region mutation and HBeAg status, HBV replication and liver function were surveyed in chronic hepatitis B patients. HBV DNA was isolated from fifteen serum samples of seven patients with chronic hepatitis B, and the X precore region was amplified by PCR and sequenced. Each sequence was compared with the consensus sequence of each patient. The most frequently observed mutations were A to T at nt. 1762 and G to A at nt. 1764 in the core promoter region and G to A at nt. 1896 in the precore region. These mutations were more commonly found in HBeAg-negative serum samples than in HBeAg-positive (p < 0.001, respectively). Of 7 serum samples with the wild type or wild/mutant mixed type in the precore region, 6 were HBeAg-positive and had a high HBV DNA polymerase (DNA p) activity and serum alanine aminotransferase (ALT) elevation. On the other hand, 6 serum samples with the mutant type only were HBeAg-negative and had a low DNA-p activity and normal ALT level. Interestingly, 2 serum samples with the mutant type only had a high DNA-p activity and serum ALT elevation, despite being HBeAg-negative. Core promoter region mutation was not associated with HBV markers or serum ALT level. In conclusion, mutation at nt. 1896 in the precore region causes seroconversion from HBeAg-positive to -negative and is accompanied by reductions of viral replication and hepatitis activity. PMID- 11494583 TI - [Visual memory impairment during postpartum]. AB - Using a visual memory test we studied memory in pregnant women. Benton's Visual Retention Test was used with 54 pregnant women three times: at term, 3-4 days post-partum, and one month after birth. There was a significant decline in visual memory at 3-4 days post-partum (p < 0.01). There was no interaction between hours of sleep and memory of pregnant women at any period. Although memory declined at 3-4 days, there was no interaction between this decline and duration of labor, intra-partum hemorrhage, or memory at one month post-partum. It is not yet clear why visual memory declined shortly after birth. Further studies on post-partum memory decline as well as the possible relationship to hormonal and mood changes are necessary. PMID- 11494584 TI - Sporadic amyotrophic lateral sclerosis showing abnormal somatosensory evoked potentials: a report of three cases. AB - OBJECTIVE: Although dysfunction of the sensory systems in sporadic amyotrophic lateral sclerosis (ALS) has been reported, the clinical characteristics of such cases still remain unknown. We therefore performed a clinico-electrophysiological analysis of sporadic ALS patients. MATERIAL AND METHODS: Twelve ALS patients (aged 36-66 years), who had their somatosensory evoked potentials (SEPs) evaluated, were reviewed and their clinical characteristics were delineated. In addition, needle EMG, sensory nerve conduction velocities, motor evoked potentials (MEPs) and cervical MRI or plain X-ray of the neck were also recorded. RESULTS: Three cases were segregated from the other 9 patients because of predominant upper motor neuron signs with pseudobulbar palsy and abnormal posterior tibial nerve and/or median nerve SEPs. The MEPs were also abnormal in these 3 patients and the brainstem auditory evoked potentials were abnormal in one patient. EMG revealed less involvement in the lower motor neurons. CONCLUSION: Sporadic ALS with a predominant upper motor neuron sign and also demonstrating pseudobulbar palsy with abnormal SEPs, may therefore form a clinical subtype of ALS. PMID- 11494585 TI - [Changes of fatty acid composition of brain synaptosomal phospholipids in isoflurane exposure rat]. AB - We examined effects of inhalational anesthetic drug, isoflurane, on phospholipid and fatty acid in brain synaptosome. Wistar strain male rat was treated by inhalation of isoflurane. Rats were divided into 3 groups each 6 rats, one was 1 minimum alveolar concentration (MAC) exposure group, 1MAC group, and another was 2 times of MAC exposure group, 2MAC group, and the other was non exposure group, control group. The animals were kept in to box (0.343 m3) and the gas flow rate was set in 4 L/min by anesthetic instrument. After 60 minutes of exposure, rats were decapitated. Immediately, cerebrums were removed and fraction of synaptosome was sampled. In 2MAC group, C14:0 of phosphatidylcholine (PC) increased significantly as compared to the control group, but C16:1, C18:0, C18:2 and C20:3 decreased significantly. And also, C18:2 and C20:3 decreased significantly in 1MAC group. In terms of phosphatidylethanolamine(PE), C18:1 in 1MAC group, C14:0 and C16:1 in 2MAC group increased, but C20:3, C20:4 and C22:5 in 2MAC group decreased significantly as compared to the control group. Regarding phosphatidylserine + phosphatidylinositol, C14:0 in 2MAC group increased, but C22:5 decreased. In lysophosphatidylcholine, C12:0 and C14:0 in 2MAC group and C18:0, C20:4 in 1MAC group increased significantly, but C18:1 in 2MAC decreased. The changes of phospholipids and fatty acid in synaptosome were due to the metabolism of phospholipids of basic matrix and this was caused by effects of isoflurane on neural cellular membrane. The results indicated the suppression of membrane activity. Isoflurane has physiological activity on metabolism of phospholipid of cellular membrane. Thus, it has effects on neural cellular functions in brain. PMID- 11494586 TI - Influence of ethanol on metabolism of methamphetamine in rats including hair analysis. AB - To investigate possible influences of ethanol (EtOH) on metabolism of methamphetamine (MA), dark agouti (DA) rats were assigned to 3 groups including EtOH mono-dosing group, MA mono-dosing group and combined dosing group of EtOH and MA. Following successive administration of respective drugs, time-course changes of plasma and urine levels of EtOH and its metabolites, as well as alterations of plasma, urine, hair levels of the MA and its metabolites, were monitored. Preliminary hydrolysis of the glucuronate conjugates was required because parahydroxymethamphetamine (p-OHMA) and parahydroxyamphetamine (p-OHAP) were promptly conjugated with glucuronic acid in the body. When compared with EtOH group, the maximum plasma concentration of EtOH occurred with some delay in EtOH + MA group, suggesting possibility of some delay of its metabolism. MA concentrations in EtOH + MA group were slightly higher until 6 hours after administration when EtOH was detectable in the blood whereas concentrations of amphetamine (AP) in EtOH + MA group were less, on the contrary. Concentrations of p-OHMA and p-OHAP were obviously less in EtOH + MA group. These findings indicate that N-demethylation and parahydroxylation of aromatic ring of MA would be inhibited in the presence of EtOH. Concentrations of MA and AP in hair were higher in EtOH + MA group. Not only difference in AUC but also possible increase in uptake of the drugs into hair could likely account for this phenomenon. PMID- 11494587 TI - [The total-quality-management-analysis of the continuation and discontinuation of alcohol family treatment: a grassroots approach]. AB - A family support/treatment program was provided to thirty-three cases where a drinking family member (identified patient) had shown alcohol related problems but not yet admitted the problem. After a period of between thirteen and twenty one months of family treatment, fourteen (42.4%) identified patients started their own treatment. The only statistically significant factor that was related to the patients' treatment program participation was the continuation of family members' involvement in a family treatment program. Among the thirty-three cases, a little less than one half (48.5%) continued the family program. In order to increase the patients' participation, it is crucial to encourage family members to continue their family support/treatment program. In order to identify factors that contribute treatment continuation as well as dropouts, workshops were held with those who dropped out and those who continued the family treatment program. The Total-Quality-Management (TQM) affinity and arrow diagram techniques were employed to classify the participants' statements and to find cause-effect relationships among the identified factors, respectively. Five family treatment discontinuation factors were identified: 1) a lack of information about family support program, 2) resistance against a "family change" orientation in family treatment program, 3) family member burnout, 4) a misfit between family needs for immediate problem solutions and what family program offers, and 5) a temporal improvement of patients' drinking problems. While widely varied factors were found to contribute discontinuation, only a very few factors were identified to facilitate the treatment continuation. It was concluded that treatment discontinuation, rather than continuation, was the norm among the families of problem drinkers. Based on the above findings, three kaizen plans were proposed. First, in order to make sure that family members obtain necessary information about the family support/treatment program, a pamphlet would be created and handed out to those who come to family treatment. Second, family support efforts would be emphasized more. Treatment staff is expected to become more cautious with regard to the family behavior change facilitation, especially at the early stage. Third, treatment staff is expected to become more aggressive about contacting family group members when they do not show up to a meeting. PMID- 11494588 TI - [The Denial and Awareness Scale (DAS)]. AB - Focusing on denial of drinking problems and raising awareness are two important treatment tasks at the initial stage of alcoholism treatment. However, an objective and clinically practical measure to assess patients' and family members' denial and awareness of alcohol-related problems has not yet been available. The DAS (Denial and Awareness Scale by Aro Ino and Shigeo Tatsuki) was developed in order to predict prognosis and evaluate treatment outcomes of alcoholism patients. The preliminary (DAS-1) patient scale was administered to 370 alcoholism patients. Their family members responded to the DAS-1 family member subscales. The abstinence period was then triangulated among the patient, a family member, and treatment staff members. For the purpose of the current study, 218 responses were used for item analyses and prediction of abstinence periods. Stepwise regression analyses of preliminary (DAS-1) sub-scales demonstrated that several sub-scales significantly predicted abstinence periods. After factor-analytic item re-classification of DAS-I items, DAS-2 was constructed. DAS-2 sub-scales were then regressed onto abstinence periods by a backward stepwise regression method. The analysis using the entire complete sample (N = 189) where both patients and their family members' responses were available demonstrated significantly that "Patient's Awareness" increased abstinence periods while "Family Troubled Feelings" decreased it, and that "Patient's Awareness" also galvanized "Changes in Patient's Mindset". Additional analyses were conducted in order to examine if predictive values of DAS-2 subscales differ according to varying stages of abstinence periods among the study subjects. The results showed that different sets of subscales predicted the abstinence periods depending on the treatment stages, i.e., abstinence within 1 year, 2 years, 3 years, and 4 years. These findings corresponded with the authors' clinical experiences. The current study produced several regression prediction equations utilizing the DAS-2. Further evaluation studies shall examine the effectiveness of the DAS-based prediction/prognosis of the abstinence period of alcoholism patients. PMID- 11494589 TI - [A review of epidemiological studies on the relationship of social networks and support to depressive symptoms in the elderly]. AB - This paper reviews evidence concerning the relationship of social support and networks to psychological well-being in elderly people. Although few studies have used comparative concepts and featured measurements of social networks and social support, the overall findings are that depressive symptoms in elderly people are associated with (a) smaller size of social networks which are supportive, (b) lower emotional support which is anticipated or received, (c) lower instrumental support which is anticipated, and (d) not providing support to others, and that depressive symptoms are greater when the amount of received instrumental support is larger. There is evidence consistent with both direct effects of social networks and support on reducing depressive symptoms, and buffering effects to protect persons from adverse effects of stressful life events. In addition, physical health status, activities of daily living, income, and marital status are associated with depression among the elderly. A well-conducted longitudinal study is essential for research in this area in Japan. PMID- 11494590 TI - [Risk factors for infant mortality from diseases in Japan 1995-98]. AB - OBJECTIVE: Since 1995, additional information (i.e. birth weight, singleton/multiple births, gestational weeks, maternal age, maternal parity and stillbirth experience) has been required for certificates of infant (less than 1 year of age) death from diseases in Japan. The present study examined the effects of biological, demographic and social variables, as reported on birth and death certificates, on infant, neonatal and postneonatal mortality in Japan. METHODS: Using data from vital statistics between 1995 and 1998, more than 4,787,000 livebirths and 16,000 infant deaths from diseases were analyzed. Univariate and multivariate analyses with the Poisson regression model were employed to assess the effects of variables on infant, neonatal and postneonatal mortality by singleton and multiple livebirths separately. RESULTS: The infant mortality rates from diseases were 3.2/1000 for singleton livebirths and 17.7/1000 for multiple livebirths. In singleton livebirths, low birth weight, infant born in earlier years, being a male infant, employment status as "unemployed or unknown", short gestational weeks, late birth in multiparity and maternal stillbirth experience were all significantly related to increased risk of neonatal and postneonatal deaths. Teenage mother were also at high risk of postneonatal deaths. Regional differences were observed. Compared with singleton livebirths, birthweight specific mortality rates in multiple livebirths were relatively low among infants weighing under 2500 g. In multiple livebirths, elevated risk of death was associated with low birth weight, infant born in earlier years, employment status as "unemployed or unknown" and short gestational weeks. However, late birth in multiparity was related to a reduced risk of death, and maternal stillbirth experience was not a significant variable. CONCLUSION: This study provided the first quantitative estimate of risk of infant mortality from diseases in Japan. Since a more detailed elucidation of actual conditions and risk factors of infant deaths by vital statistics has become possible, efficient measures for improvement of infant mortality are to be expected. PMID- 11494591 TI - [Factors allowing continuous working of female nurses in the People's Republic of China--survey of 15 hospitals in 5 areas]. AB - PURPOSE: Few investigations have been conducted on working, childcare and home education among female nurses (nurses) in the People's Republic of China (P.R. of China). The purpose of this study was to clarify the factors allowing continued working of female nurses. METHODS: The subjects were nurses from 15 hospitals in 3 provinces and 2 autonomous areas. They were surveyed by questionnaire regarding their childcare and home education. In was distributed by the Ministry of Health in the P.R. of China and the data were collected between February and April in 1996. RESULTS: A total of 4,284 (80.0%) questionnaires were collected. 1. About 60% of the nurses began working when they were 15 to 19 years old. The average length of service was 13.9 years. 2. Their professional position correlated directly with their years of service. 3. 4.3% of them had not continued working. 4. The nurses entrust their children to day-care institutions (61.2%) or grandparents (22.3%) during the daytime, and to the father (66.3%) or the grandparents (19.5%) during the night. 5. 17.6% of them have experienced being away from home for more than one month due to their job. 6. 66.8% of the nurses have experienced more than one month official trip, and had children under 14 years old at the time. They entrusted their children to day-care institutions (34.1%), to the father (29.2%) and grandparents (25.3%) during the official trip. CONCLUSIONS: In summary, most nurses are employed when they are young and go on working as nurses. It is possible for them to continue their work after marriage, childbirth, and while they are bringing up their children. The high rate of working woman, sufficient nursery schools and family cooperation in housework can be pointed out as main factors that enable them to continue their work. PMID- 11494593 TI - [Utilization of Maternal and Child Health handbook in Japan]. AB - OBJECTIVES: In Japan, Maternal and Child Health (MCH) handbook have been widely used for more than fifty years. However, there has been no evaluation research from the users' point of view. We therefore conducted a questionnaire survey of guardians to evaluate how they utilize MCH handbooks. METHODS: A well-structured questionnaire survey was carried out in 231 municipalities, towns and villages of four prefectures (Gifu, Hiroshima, Niigata and Shizuoka) and one city (Yokohama) in November and December 1999. The targets were 13,271 guardians who visited health stations for 18-month examinations of their children and agreed to participated in our research. The questionnaire covered situation of usage and loss by guardians, utility from the users' perspective, and suggestions for improvement. RESULTS: We obtained answers from 10,900 guardians. As for reading rate and writing rate, 98.3% of respondents had read and 97.8% of them had written down something in relation. Only 0.9% of respondents had lost this MCH handbook. Generally, 87.0% of respondents answered that MCH handbook was helpful for child bearing and 81.6% of them said the record for immunization was useful. However, 34.1% of respondents answered it was not simple to utilize MCH handbook and 60.6% of them requested more detail on child bearing. As for dental health, the completion rate for information was low and only 21.3% of respondents reported for the dental record was useful. CONCLUSIONS: The research shows MCH handbook are highly utilized in Japan by almost all guardians. However, there are still problems to be solved; many guardians feel that they are not simple to use and the section on dental health is not highly particularly helpful. Improvements should be made in future in light of the viewpoints of guardians and parents. PMID- 11494592 TI - [Childcare and home education in families of female nurses in the People's Republic of China--survey of 15 hospitals in 5 areas]. AB - PURPOSE: The purpose of this study was to investigate the actual conditions of childcare and home education among families of female nurses in the P. R. of China. METHODS: The subjects were nurses from 15 hospitals in 3 provinces and 2 autonomous areas. They were surveyed by questionnaire regarding their childcare and home education. It was distributed by the Ministry of Health in the P. R. of China and data were collected between February and April in 1996. RESULTS: A total of 4284 (80.0%) questionnaires were collected. 1. The age of the subjects was between 18 and 62 years old with a mean of 32.9 +/- 9.0 (SD) years 71.4% of them had husbands, whose ages were between 23 and 71 years old, the mean age being 38.3 +/- 8.4 (SD) years. The family types were 63.2% nuclear family and 33.7% extended family, with 3.1% being single. 65.1% of the subjects had children, whose mean number was 1.1 +/- 0.4 (SD). Firstborn children were 49.9% girls and 50.1% boys. 2. The most popular method of infant nutrition was breast feeding, utilized by 60.1%. The highest rate (67.8%) was in the age group of 25 to 29 years old (P < 0.01). 3. The practice of "swaddling" (wrapping the child so as not to allow movement) was more common in the over 40 year old age group than the under 40 year old group (P < 0.01). Swaddling-practice showed significant differences by area. 4. The rates for children who helped with housework were 50.3% for girls and 46.7% for boys. 5. A number of children between 7 and 18 years old were studying English privately. 6. The greatest expectation of the subjects, for their children aged between 7 and 18 years old, was to study. CONCLUSIONS: 1. Infant nutrition showed significant differences between generations, and swaddling-practice differed with the generation and the area. 2. Although girls help their parents more than boys in Japan, boys and girls equally helped their parents in the P. R. of China. 3. Concerning the topics of private learning and parents expectations, the results were similar to those in Japan. PMID- 11494594 TI - [Synthesis and its application to the synthesis of biologically active natural products of new and versatile chiral building blocks]. AB - This article describes a design and synthesis of new and versatile chiral building blocks and its application to the biologically active natural product synthesis. The chiral building blocks were prepared using a biocatalysis in an enantiomerically pure state. As an application of the above chiral building blocks to the synthesis of biologically active natural product, we demonstrated the diastereodivergent synthesis of the 3-piperidinol alkaloids cassine, spectaline, prosafrinine, iso-6-cassine, prosophylline, prosopinine, and also established the flexible route to the 5,8-disubstituted indolizidine or 1,4 disubstituted quinolizidine type of Dendrobates alkaloids. As another application to the synthesis of biologically active alkaloids, we accomplished the first enantioselective total synthesis of marine alkaloids clavepictines A, B, and pictamine using a highly stereoselective Michael type quinolizidine ring closure reaction as the crucial step, and the first total synthesis of a marine alkaloid lepadin B was also achieved using aldol cyclization controlled by a A strain. PMID- 11494595 TI - [Maturation pathway of hemolysin of Aeromonas sobria and the mechanism of action of the hemolysin]. AB - Aeromonas sobria has been recognized as pathogens associated with acute gastroenteritis in both adults and children. The major virulence factor has been proposed to be hemolysin which possesses both hemolytic and enterotoxic activities. Mature (bio-active) hemolysin secreted out of cells binds to the target cells of the host and injure the cells. However, hemolysin remained in bacteria can not express such toxicity. It means that the maturation and secretion pathway of hemolysin is closely related to the pathogenicity of bacteria. Therefore, I examined the pathway and clarified the following events. Hemolysin synthesized in cytoplasm translocates across the inner membrane and appears in a periplasmic space. Hemolysins appeared in the space associates to form dimer in the space. The C-terminal region of hemolysin functions as a trigger in the association. Dimerized hemolysin crosses the outer membrane and emerges in milieu, but monomer can not cross it. Therefore, the C-terminal region of hemolysin attributes not only to the formation of the dimer but also to its secretion into milieu. Hemolysin emerged in milieu is inactive. Inactive hemolysin is converted to bio-Active hemolysin by deleting its carboxyl-terminal 42-amino-acid peptide. Active hemolysin generated binds to the receptor of the target cell and stimulates the production of cyclic AMP by the cell. I assume that this stimulation closely relates to the induction of diarrhea by hemolysin. PMID- 11494596 TI - [Synthetic studies of the bioactive natural products by using the thermal electrocyclic reaction of 6 pi electron systems]. AB - We are developing the synthesis of biologically interesting condensed heteroaromatic compounds, including natural products by the thermal electrocyclic reaction of 6 pi electron system incorporating the double bond of the principal aromatic or heteroaromatic ring. In this report, we describe three types of electrocyclic reactions as follows; 1) the synthesis of highly-substituted carbazole alkaloids based on the allene-mediated electrocyclic reaction involving an indole 2,3-bond, 2) the synthesis of beta-carboline alkaloids and isoquinoline 5,8-quinone alkaloids based on the thermal electrocyclic reaction of an 1 azahexatriene system involving an indole 2,3-bond or benzene 1,2-bond, and 3) the synthesis of new tetracyclic pyrido[2,3-b]indole, grossularines, based on the thermal electrocyclic of an 2-azahexatriene system including the indole 2,3-bond. PMID- 11494597 TI - [Studies on the constituents of Cimicifuga species]. AB - The chemical constituents of Cimicifuga simplex and its related species were reexamined using HPLC and high resolution spectral analysis. From C. simplex, C. acerina and C. japonica, a new alkaloid, 59 new cycloartane triterpene glycosides and 11 new aromatic constituents were isolated with the previously reported compounds. The latter aromatic constituents were identified in HPLC analysis of the extracts from crude drugs due to C. dahurica, C. heracleifolia and C. foetida. Several interesting topics in this study are reviewed about separation of unstable and closely related compounds by HPLC, rearrangement reactions, chemical conversion for confirmation of absolute configuration, isomerism in NMR solutions, X-ray crystal analysis, spectral analysis of complicated structures, isolation of glycoside malonates, high yield of enzymatic hydrolysis, specific CD curve due to a cycloart-7-ene system, and so on. Biological activities of a major glycoside, cimicifugoside, a modified triterpene, acerinol, and benzyltartaric acid derivatives such as fukinolic acid and cimicifugic acids were also mentioned. PMID- 11494599 TI - [Chemistry and biosynthesis of prenylflavonoids]. AB - Many isoprenylated flavonoids have been isolated from mulberry trees and related plants (Moraceae). Among them, kuwanons G (13) and H (14) were the first isolated active substances exhibiting a hypotensive effect from the Japanese Morus root bark. These compounds are considered to be formed through an enzymatic Diels Alder reaction of a chalcone (15) and dehydro-kuwanon C (16) or its equivalent. Since that time, about forty kinds of Diels-Alder type adducts structurally similar to that of 13 have been isolated from the moraceous plants. Some strains of Morus alba as well as M. bombycis callus tissues have a high productivity of mulberry Diels-Alder type adducts, such as chalcomoracin (26) and kuwanon J (28). The biosynthesis of the mulberry Diels-Alder type adducts has been studied with the aid of the cell strain. Chalcomoracin (26) and kuwanon J (28) were proved to be enzymatic Diels-Alder type reaction products by the administration experiment with O-methylchalcone derivatives. Furthermore, for the isoprenoid biosynthesis of prenylflavonoids in Morus alba callus tissues, a novel way through the junction of glycolysis and pentose-phosphate cycle was proposed. The crude enzyme fraction catalyzing the Morus Diels-Alder type reaction could be isolated. Studies of phenolic constituents of licorice (Glycyrrhiza species) were carried out. On the course of the structure determination of the phenolic constituents of licorice, two new NMR structure determination methods for prenylflavonoids were found. Furthermore, the prenylphenols isolated from licorice were summarized according to the origin of the materials. PMID- 11494598 TI - [Molecular science of the living organism: the case of G-proteins]. AB - The concept of Molecular Science of the Living Organism was described, where the living state is explained as the purposive flows of the quantum mechanically controlled chemical reaction systems which support the homeostasis of the living organism. In the 21st century, the post genomic sequence era, the concept may be a self-evident truth. Molecular Science of the Living Organism was presented in the case of G-proteins: i.e., the atomically controlled mechanism of 1. the carcinogenesis which originates from the point mutation of ras p21, 2. the activation of a receptor protein at the cell membrane, especially in the case of bacteriorhodopsin, 3. the activation of an inactive G-protein by the activated receptor protein. PMID- 11494600 TI - [Studies on the mechanism of subcellular distribution of basic drugs based on their lipophilicity]. AB - This paper described the studies on the mechanism of subcellular distribution of lipophilic weak bases. Although the tissue distribution of basic drugs appeared to decrease with time simply in parallel with their plasma concentration, their subcellular distribution in various tissues exhibited a variety of patterns. Basic drugs were distributed widely in various tissues, but were concentrated in lung granule fraction, where their accumulation was dependent on their lipophilicity and lysosomal uptake. As the plasma concentration of drugs decreased after maximum level, the contribution of lysosomes to their subcellular distribution increased. The uptake of the basic drugs into lysosomes depended both on their intralysosomal pH and on the drug lipophilicity. As the lipophilicity of the basic drugs increased, they accumulated more than the values predicted from the pH-partition theory and raised the intralysosomal pH more potently, probably owing to their binding with lysosomal membranes with or without additional intralysosomal aggregation. These phenomena should be considered as a basis of drug interaction in clinical treatments. PMID- 11494601 TI - A parent's perspective: providing compassionate and effective care. PMID- 11494602 TI - Tapping Disney inspiration: transforming healthcare. PMID- 11494603 TI - Collaborating for excellence: taking academia to a rural setting. PMID- 11494604 TI - Shared mental healthcare: a collaborative consultation relationship. The North York General Hospital experience. PMID- 11494605 TI - Hospital mergers: why some fail and others succeed. PMID- 11494606 TI - Trends in public opinion on healthcare worth watching. PMID- 11494607 TI - Changes in Ontario's core mental health care. PMID- 11494608 TI - Towards the virtual hospital: an ecological approach to network development in health services. PMID- 11494609 TI - Development of a new telementoring program. PMID- 11494610 TI - The British Columbia Cancer Agency: a comprehensive and integrated system of cancer control. PMID- 11494611 TI - Insurance considerations for magnetic resonance imaging installations. PMID- 11494612 TI - A genetic revolution in healthcare. PMID- 11494613 TI - A report card on health(care) report cards. PMID- 11494614 TI - The truth about case presentation. PMID- 11494616 TI - Esthetic crown lengthening. PMID- 11494615 TI - Strategies for complete denture success: beyond technical excellence. AB - Many patients are quite satisfied with dentures that are extremely poor, while other patients are dissatisfied with dentures that are technically excellent. Overall, some 25% of patients are likely to be dissatisfied with their artificial teeth. This article addresses the aspects of denture success not covered by technical considerations. The strategies are based on the principles of good communication and involve an understanding and respect for patients' motivations in wearing dentures. The strategies include help in the development of neuromuscular control and in training regarding mastication. Most importantly, they emphasize patient involvement in the development of the denture. Patients must be made aware of their responsibilities in achieving a satisfactory outcome in the complex patient/denture relationship. PMID- 11494617 TI - Improvement of oral health in the postgenomic era: opportunities for government/industry partnerships. PMID- 11494618 TI - Perceived malocclusion and other teeth-associated signs and symptoms in temporomandibular disorders. AB - This article reports the results of a clinical study that evaluated adjunctive teeth-associated signs and symptoms before and after nonsurgical temporomandibular disorder therapy. Eighty-nine patients were referred to a private prosthodontic practice in the Washington, DC metropolitan area. Of the 89 patients, 75 were selected for the clinical study after completing the interview, history, and clinical examination. The major signs and symptoms were recorded, along with adjunctive teeth-associated signs and symptoms. The treatment included an anterior programming device, a centric relation occlusal device, and, when indicated, a selective occlusal equilibration. The results were recorded after five treatment visits. Of 75 patients, 67 made good-to-excellent improvement in the signs and symptoms of their major temporomandibular disorder. Patients with adjunctive teeth-associated signs and symptoms demonstrated a marked reduction in these signs and symptoms as assessed by their mental perception and verbal feedback. The symptoms of perceived awareness of malocclusion were not resolved with centric relation occlusal device therapy in 22 of the 24 patients with this symptom. However, excellent improvement was noted in this symptom when occlusal device therapy was followed by a selective occlusal equilibration. Thirty-eight patients in this study had difficulty in making lateral gliding articulation movements with the occlusal device out of the mouth. This problem was resolved in 36 patients after removal of eccentric occlusal interferences. It is suggested that adjunctive teeth-associated signs and symptoms be initially recorded and addressed in patients, especially if extensive or invasive dental therapy is planned. Although this clinical study in a private practice was carefully performed, it does not meet the criteria of evidence-based research, because a control group was not included. PMID- 11494619 TI - Maxillofacial implications and surgical treatment of arthrogryposis multiplex congenita. AB - A case of arthrogryposis multiplex congenita that affected a patient's temporomandibular joint is described. Preoperatively, the patient's interincisal opening was limited to 9 mm. Elective surgery was performed, which consisted of bilateral coronoidotomies, right and left meninscectomies, capsular release, and lateral pterygoid myotomies. Physical therapy was initiated postoperatively. Eighteen weeks after the surgery, the patient was able to open 18 mm and force open to 20 mm. The patient also noted significant improvement in speech and jaw function in the postoperative period. PMID- 11494620 TI - Pit-and-fissure sealant materials. PMID- 11494621 TI - Comparative seven-day clinical evaluation of two tooth whitening products. AB - A 1-week study was conducted to compare the tooth whitening efficacy of two carbamide peroxide-based products (one containing 5% carbamide peroxide and one containing 10% carbamide peroxide). In addition, the perception of transient tooth hypersensitivity associated with the use of these products was subjectively evaluated. Sixty participants took part in a double-blind, randomized, parallel clinical study. Change in tooth color was measured by chroma meter and Vita shade guide at the initiation of the study and after 7 days of product use. Color change (delta E) was calculated using the color-difference equation established by the Commission Internationale de L'Eclairage. Data concerning dental hypersensitivity was assessed by subjective panelist questionnaires administered after the day 7 recall examination. Results of the whitening data showed that there was no significant difference between the two products. After 1 week, the mean delta E for the product containing 5% carbamide peroxide was 4.43 +/- 1.89 and the mean delta E for the product containing 10% carbamide peroxide was 4.05 +/- 1.84. However, the subjective data collected on tooth hypersensitivity showed that the product containing 5% carbamide peroxide was associated with less discomfort. Of the group using the 5% carbamide peroxide product, 20% reported transient sensitivity of their teeth after product use for 1 week compared with 53% of the group using the product with 10% carbamide peroxide. Statistical analysis using a 2-tailed t test showed significant differences between the two groups (P < .05). The data suggest that these products are clinically equivalent for tooth whitening; however, the product containing 5% carbamide peroxide was associated with less tooth hypersensitivity after 1 week of application. PMID- 11494623 TI - [Small or big]. PMID- 11494622 TI - Esthetic changes using minimal preparations and new composites. PMID- 11494624 TI - [Coronary angioplasty in the acute phase of myocardial infarction in a low-volume center]. AB - The efficacy of coronary angioplasty in the treatment of acute myocardial infarction was assessed in a low volume centre. Between January 1994 and May 1999, 148 consecutive patients (mean age 59 years, 81% men) with acute myocardial infarction, admitted within 12 hours, were included in this retrospective analysis. On admission, 14% of patients were in cardiogenic shock. The average time between the onset of chest pain and arrival at hospital was 244 +/- 183 mins. Reperfusion (TIMI 3 flow) was obtained on average 111 +/- 60 mins after arrival at hospital and 81 mins after informing the on-call team. After angioplasty, residual stenosis < 50% was obtained in 91% of cases. TIMI 3 flow was obtained in 85% of cases (TIMI 2 + 3 in 93% of cases). Over the years, the delay before treatment decreased and the results of angioplasty improved. In the last 79 patients, residual stenosis < 50% was obtained in 95% of cases, TIMI 3 flow in 87% of cases (TIMI 2 + 3 in 97% of cases). The stenting rate increased from 16% before 1997 to 61% thereafter. The hospital mortality was 4%. Direct or salvage angioplasty in the first 12 hours of myocardial infarction in some low volume centres may be carried out safely with intervention times and success rates comparable to those reported in the literature. PMID- 11494625 TI - [Results of coronary artery surgery in octogenarians]. AB - Over an 11 year period from January 1990 to December 2000, 3282 patients underwent isolated or combined surgical myocardial revascularisation. In this group, 42 were aged 80 or over (maximum 87 years), 1.3% of the total patient population. The mean age of this subgroup was 81.8 +/- 1.75 years) with a male predominance (61.9%). All patients were autonomous and considered to be in good general and psychological health. Preoperative coronary angiography showed 33.3% of left main stem lesions either alone or associated with a right coronary lesion. The ejection fraction was over 50% in 78.6% of cases. Saphenous vein grafts were used in all but 5 patients who also had left internal mammary artery grafts. Thirteen patients (31%) underwent combined valvular surgery (11 aortic and 2 mitral valve) and 2 patients underwent combined vascular surgery. Three patients were operated as an emergency. A total of 5 patients died in the first 30 postoperative days, a hospital mortality of 11.9%. There were 2 postoperative hemiplegias and 2 cases of renal failure which were aggravated in the postoperative period. The other patients were discharged from hospital with a satisfactory cardiac and functional status. The global mortality was 14% at 3 years and 18% at 5 years. The main bad prognostic factor for survival was the association of aortic valve surgery. In selected octogenarians in good general and psychological health without severe co-morbid conditions, surgical myocardial revascularisation may be considered with an acceptable operative risk. PMID- 11494626 TI - [Modeling of coronary arteries and cardiovascular risk factors]. AB - The aim of this study was to assess the impact of cardiovascular risk factors on the modelling of atherosclerotic coronary arteries. One hundred consecutive patients who underwent coronary angioplasty were studied by endocoronary ultrasonography at the site of dilatation. At the site of the treated stenosis of the dilated arteries there was either compensatory widening or positive modelling (PM), or focal contraction or negative modelling (NM) if the total surface area (TSA) of the artery at the site of dilatation was greater or smaller than the total surface area of the proximal or distal reference segments. PM was observed in 53 cases (53%) and NM in 47 cases (47%). Lesions with NM had smaller TSA (13.7 +/- 5.8 versus 20.8 +/- 6.4 mm2, p < 0.0001) and a smaller atheromatous plaque (11.8 +/- 5.6 versus 19.1 +/- 6.5 mm2, p < 0.0001) than lesions with PM. Cardiovascular risk factors such as hypercholesterolaemia, smoking and hypertension were not predictive of either form of arterial modelling and there was no relationship between the cardiovascular risk factors and the qualitative appearances of the plaque studied. PMID- 11494628 TI - [Myocardial bridge: description, significance, treatment]. AB - The myocardial bridge is a well known anomalous trajectory of the coronary arteries, the significance of which has been discussed for a long time. It now appears that some myocardial bridges may cause myocardial ischaemia with clinical electrocardiographic, scintigraphic or metabolic signs--even myocardial infarction or sudden death. In these symptomatic cases, treatment is usually medical and rarely surgical. In the last few years, angioplasty and stenting have been used more frequently in cases resistant to medical therapy and appear to be an effective alternative to surgery. PMID- 11494627 TI - [Distribution and treatment of cardiovascular risk factors in coronary patients: the Prevenir Study]. AB - The aim of this study was to evaluate the distribution and hospital treatment of cardiovascular risk factors in coronary patients. A transverse observational study was carried out in 77 cardiological centres throughout France. All patients with a history of myocardial infarction or of unstable angina during the month of January 1998 were enrolled. The clinical features of 1334 patients (71.4% men, 746 myocardial infarction, 588 unstable angina) on hospital admission were analysed. The prevalence of smoking, dyslipidaemia, hypertension and obesity was 49, 45, 42, 19 and 11% respectively in the men and 17, 46, 63, 23 and 10% respectively in the women. The number of treatable risk factors slightly decreased in the oldest age group. The prevalence of hypertension increased with age whereas smoking and dyslipidaemia decreased in both men and women. Obese and diabetic patients had more risk factors than the others. A little less than half of patients with dyslipidaemia were under no preventive measures (diet and/or lipid-lowering drugs) and 40% of men with a previous history of coronary artery disease continued to smoke. The authors conclude that men over 85 and women over 75 years of age have fewer risk factors than other age groups and the type of risk factor varies with age. The treatment of dyslipidaeamia and smoking is still inadequate and should be improved. PMID- 11494629 TI - [Hormone replacement therapy of menopause, heart and blood vessels]. AB - The incidence of coronary heart disease (CHD) is lower in premenopausal women than in men and post-menopausal women of the same age. The higher CHD rate after menopause is currently attributed to estrogen deficiency: many epidemiological (case-control and prospective) studies have reported a reduced risk (0.5-0.63) of CHD in post-menopausal women receiving hormone replacement therapy (HRT). Moreover, estrogens have multiple effects that would be expected to be cardioprotective, including favorable changes in lipids, endothelial function, vascular reactivity and blood flow. However, the observational studies are subject to several biases that could falsely elevate the apparent benefit of estrogens: women taking estrogens tend to be wealthier, more educated and healthier than untreated women. The american HERS (Heart and Estrogen-progestin Replacement Study; 2.763 women) is a large multicenter randomized study of secondary prevention, designed to evaluate the efficacy of HRT. Results are disappointing, since no reduced risk was observed, and the risk of CHD was even higher in women receiving HRT during the first year: 1.52 (CI 95%: 1.01-2.29). In HERS study, the treatments consisted of conjugated equine estrogens and the synthetic progestin medroxyprogesterone acetate (MPA) which are rarely used in Europe. Indeed, the effects of HRT are not equivalent depending on the dose, the route of administration, the type of progestogen. It should be emphasized that MPA, contrarily to progesterone, inhibits the beneficial effect of estrogens on lipids and experimental atherosclerosis. The route of administration of estrogens is also involved: estrogens alter hemostasis factors, and when orally administered, they have a first pass liver effect, which favors hypercoagulability. It is therefore urgent that Europeans undertake a European "HERS study" in order to investigate the possible beneficial effect of non oral estrogens (gel or patch) associated with natural progesterone. PMID- 11494630 TI - [Real time 3D echocardiography]. AB - Three-dimensional representation of the heart is an old concern. Usually, 3D reconstruction of the cardiac mass is made by successive acquisition of 2D sections, the spatial localisation and orientation of which require complex guiding systems. More recently, the concept of volumetric acquisition has been introduced. A matricial emitter-receiver probe complex with parallel data processing provides instantaneous of a pyramidal 64 degrees x 64 degrees volume. The image is restituted in real time and is composed of 3 planes (planes B and C) which can be displaced in all spatial directions at any time during acquisition. The flexibility of this system of acquisition allows volume and mass measurement with greater accuracy and reproducibility, limiting inter-observer variability. Free navigation of the planes of investigation allows reconstruction for qualitative and quantitative analysis of valvular heart disease and other pathologies. Although real time 3D echocardiography is ready for clinical usage, some improvements are still necessary to improve its conviviality. Then real time 3D echocardiography could be the essential tool for understanding, diagnosis and management of patients. PMID- 11494631 TI - [Recommendations of the French Society of Cardiology concerning the treatment of myocardial infarction after the acute phase. Epidemiology and Prevention Group]. PMID- 11494632 TI - [Localization of isolated atrial involvement of cardiac sarcoidosis]. AB - The authors report the first case of cardiac sarcoidosis involving only the interatrial septum and the atrioventricular node in a 30 year old man with known lymph node sarcoidosis. The diagnosis was suspected after the finding of an apparently innocent 1st degree atrioventricular block and confirmed by transoesophageal echocardiography and by IMATRON computerised tomography. Isolated atrial involvement of sarcoidosis is rare in the absence of any left ventricular disease. This explains the negative findings at transthoracic echocardiography and radio-isotopic investigations usually recommended for diagnosing cardiac sarcoidosis. PMID- 11494633 TI - [Dissection of the aorta complicated by aorto-pulmonary fistula]. AB - The authors report the case of chronic dissection of the aorta presenting with congestive cardiac failure. The diagnosis was made for the first time by transoesophageal echocardiography which showed both the dissection of the aorta and its fistulalisation into the pulmonary artery. Aortography confirmed the diagnosis. The patient underwent surgery which consisted of suture of the fistula and replacement of the ascending aorta with a prosthetic tube. The outcome was favourable after 8 months follow-up. PMID- 11494634 TI - [Reversible hypokinetic cardiomyopathy revealing severe hypocalcemia]. AB - Dilated cardiomyopathy may be primary or secondary. Although some causes are well known, such as toxic substances (alcohol, chemotherapy...) or viral infections, biochemical abnormalities are much less common. The authors report the case of a 58 year old woman with no previous history admitted to hospital for an inaugural episode of cardiac failure. The ECG showed sinus tachycardia with a long QT interval (560 mm) and a dilated hypokinetic cardiomyopathy with a left ventricular ejection fraction of 20%. The aetiological investigation showed severe hypocalcaemia (0.66 mmol/L) related to primary hypoparathyroidism. This is an important cause to remember because its treatment leads to correction of the cardiac disease, usually within weeks. PMID- 11494635 TI - Television viewing and childhood obesity. AB - Children spend a substantial portion of their lives watching television. Investigators have hypothesized that television viewing causes obesity by one or more of three mechanisms: (1) displacement of physical activity, (2) increased calorie consumption while watching or caused by the effects of advertising, and (3) reduced resting metabolism. The relationship between television viewing and obesity has been examined in a relatively large number of cross-sectional epidemiologic studies but few longitudinal studies. Many of these studies have found relatively weak, positive associations, but others have found no associations or mixed results; however, the weak and variable associations found in these studies may be the result of limitations in measurement. Several experimental studies of reducing television viewing recently have been completed. Most of these studies have not tested directly the effects of reducing television viewing behaviors alone, but their results support the suggestion that reducing television viewing may help to reduce the risk for obesity or help promote weight loss in obese children. Finally, one school-based, experimental study was designed specifically to test directly the causal relationship between television viewing behaviors and body fatness. The results of this randomized, controlled trial provide evidence that television viewing is a cause of increased body fatness and that reducing television viewing is a promising strategy for preventing childhood obesity. PMID- 11494636 TI - Obesity intervention among African-American children and adolescents. AB - Often, researchers and clinicians approach the African-American community from a deficit model with African Americans viewed as having less desirable health practices and higher disease risk; however, in developing interventions for African Americans, it is important to keep in mind positive aspects of black culture as they relate to obesity. For example, the cultural acceptance of a larger body type and less negative views toward overweight individuals need not be viewed as problematic or abnormal. In fact, it could be argued that majority culture has a dysfunctional view of body image and obesity. The fact that whites are less likely to be overweight than African Americans may stem from a value system that places undue emphasis on thinness, youth, and external beauty and a culture that imbues women with shame about how they look and what they eat. Thus, rather than holding whites and majority culture as the ideal, it may be important to incorporate the positive elements of black culture regarding body image and food rather than attempting to shift their values toward those of European Americans. How best to achieve a reduction in obesity and its medical consequences, without inducing undesirable shifts in body image and attitudes toward food, is a formidable but important challenge. PMID- 11494637 TI - Intensive therapies for pediatric obesity. AB - The intensive therapies that have been used to treat pediatric and adolescent obesity include very-low-calorie diets, pharmacotherapy, and bariatric surgery. None of these approaches have been reported in sufficient numbers of subjects who have taken part in well-designed experiments with long-term follow-up to demonstrate convincingly their true value for the treatment of pediatric obesity. As the potency of the therapy increases, so does its possible adverse consequences. Of the intensive approaches, only bariatric surgery can be said to have even small studies supporting its ability to induce long-lasting (> 1 year) effects on body weight in severely obese adolescents. The risks and benefits of intensive weight-management therapies should be weighed carefully before they are used with pediatric-aged patients. Until further controlled trials become available, intensive therapies for pediatric obesity should be considered only for children who have not responded to conventional weight-management programs but have significant complications of their obesity. Intensive approaches generally should be restricted to specialized centers that have experience with those treatments and should be carried out in the context of a comprehensive weight-management program. PMID- 11494638 TI - Airway management in the obese child. AB - Airway management in severely obese children requires familiarity with the equipment and techniques used for establishing a patent airway. Normal anatomic landmarks are obscured in these patients, making assessment a challenge. Therapy should be individualized, and because the airway is marginal in many of these patients, small insults, such as respiratory tract infection or loss of muscle tone during sleep, can result in a perilous airway. Pediatric airway charts should be maintained and reviewed frequently in emergency departments and clinics caring for these patients. Currently, the body of literature devoted to airway management in obese children is small. Inconsistencies in parameters make comparison of studies difficult. Continued, consistent reporting of airway management issues in these patients is needed. PMID- 11494639 TI - Childhood and adolescent obesity. Prevalence and significance. AB - The prevalence of children and adolescents with body mass index (BMI) of greater than 95th percentile has doubled in the last 2 decades (present prevalence is 10.9%) and there is a 50% increase in the prevalence of those with a BMI greater than 85th percentile (present prevalence is 22.0%) in the US. There are substantial risks for morbidity in obese children even before they reach adulthood. Further, if obesity in childhood persists into the adult years, the morbidity and mortality is greater than if the obesity developed in the adult. Screening using appropriate historical and physical data will reveal those children most in need of modification of weight gain. PMID- 11494640 TI - Ethnic issues in the epidemiology of childhood obesity. AB - Childhood obesity may be seen as a marker for high-risk dietary and physical inactivity practices. Recent increases in the prevalence of overweight and obesity among American children are not limited to one age, gender, or ethnic group, which suggests that unique behaviors of the members of various racial or ethnic subgroups of the population are unlikely to be the major contributing factors. Rather, it seems that environmental changes promoting increased energy intake and decreased energy output are occurring and have widespread impact on children from various backgrounds. Although no ethnic group is immune from the current shift in energy balance, differential rates of overweight seem to exist among ethnic groups. National probability samples of African-American, Hispanic, and white children in the United States provide clear evidence that white children are at lower risk for childhood overweight than are African-American or Hispanic children. Of concern is the lack of national data on the prevalence of overweight and obesity for Native-American and Asian-American groups. Also of concern is the aggregation of racial and ethnic subgroups, which may render prevalence rates meaningless. This possibility is clearly true with some surveys of weight status that combine diverse populations, such as Asians and Pacific Islanders, into one group. The high rates of obesity in African-American, Hispanic, and Native-American children are of concern. Although parental SES is associated inversely with childhood obesity among whites, higher SES does not seem to protect African-American and Hispanic children against obesity. In these groups, childhood obesity does not seem to be associated significantly with parental income and education. Health consequences of childhood obesity include a higher prevalence of type 2 diabetes and an increased risk for adverse levels of lipids, lipoproteins, and blood pressure. The effects of recently reported unprecedented levels of childhood overweight on subsequent risk for obesity in middle age are not known until future longitudinal data can be collected. It seems likely, however, that future health consequences of current early and severe childhood obesity will be staggering. Funding for adult follow-up of longitudinal studies of high-risk African American, Hispanic, and Native-American children is needed urgently to provide information on the long-term effects of childhood obesity. Halting the obesity epidemic is a formidable task, but the success in recent decades of drastically reducing childhood undernutrition offers hope and should spur similar action and leadership efforts. Promotion of efforts to reduce excess caloric intake with efforts to increase energy expenditure should receive paramount attention in the design of health programs. Given the relatively few published obesity-prevention and treatment studies that are designed to address specific cultural issues, it is important to promote the development of culturally appropriate intervention strategies that are shown to be effective among youth of diverse backgrounds. Although the dietary and activity goals will be similar, parental, family, and community messages and techniques grounded in cultural traditions and norms will be different for each ethnic group. This approach is crucial in the United States, a country with an increasingly diverse population. PMID- 11494641 TI - Biological influences on obesity. AB - Severely obese children are even more likely to have mutations in obesity genes than are severely obese adults. Thus, investigators searching for obesity genes commonly focus on children, with the result that many human obesity genes were first identified in studies of children. Although the development of obesity depends on living in an obesity-promoting environment, it also is influenced strongly by individual genetic composition. Thus, the discovery of new obesity genes provides new opportunities to identify causes of severe obesity. Finally, identification of individual causes of obesity may, in the future, provide for a safe, effective, and individualized treatment recommendation for each obese person. PMID- 11494642 TI - Family environmental factors influencing the developing behavioral controls of food intake and childhood overweight. AB - Although a large body of research has assessed direct genetic links between parent and child weight status, relatively little research has assessed the extent to which parents (particularly parents who are overweight) select environments that promote overweight among their children. Parents provide food environments for their children's early experiences with food and eating. These family eating environments include parents' own eating behaviors and child feeding practices. Results of the limited research on behavioral mediators of familial patterns of overweight indicate that parents' own eating behaviors and their parenting practices influence the development of children's eating behaviors, mediating familial patterns of overweight. In particular, parents who are overweight, who have problems controlling their own food intake, or who are concerned about their children's risk for overweight may adopt controlling child feeding practices in an attempt to prevent overweight in their children. Unfortunately, research reveals that these parental control attempts may interact with genetic predispositions to promote the development of problematic eating styles and childhood overweight. Although the authors have argued that behavioral mediators of family resemblances in weight status, such as parents' disinhibited or binge eating and parenting practices are shaped largely by environmental factors, individual differences in these behaviors also have genetic bases. A primary public health goal should be the development of family-based prevention programs for childhood overweight. The findings reviewed here suggest that effective prevention programs must focus on providing anticipatory guidance on parenting to foster patterns of preference and food selection in children more consistent with healthy diets and promote children's ability to self-regulate intake. Guidance for parents should include information on how children develop patterns of food intake in the family context. Practical advice for parents includes how to foster children's preferences for healthy foods and how to promote acceptance of new foods by children. Parents need to understand the costs of coercive feeding practices and be given alternatives to restricting food and pressuring children to eat. Providing parents with easy-to-use information regarding appropriate portion sizes for children is also essential as are suggestions on the timing and frequency of meals and snacks. Especially during early and middle childhood, family environments are the key contents for the development of food preferences, patterns of food intake, eating styles, and the development of activity preferences and patterns that shape children's developing weight status. Designing effective prevention programs will, however, require more complete knowledge than currently available regarding behavioral intermediaries that foster overweight, including the family factors that shape activity patterns, meals taken away from home, the impact of stress on family members' eating styles, food intake, activity patterns, and weight gain. The research presented here provides an example of how ideas regarding the effects of environmental factors and behavioral mediators on childhood overweight can be investigated. Such research requires the development of reliable and valid measures of environmental variables and behaviors. Because childhood overweight is a multifactorial problem, additional research is needed to develop and test theoretic models describing how a wide range of environmental factors and behavioral intermediaries can work in concert with genetic predispositions to promote the development of childhood overweight. The crucial test of these theoretic models will be in preventive interventions. PMID- 11494643 TI - The neuroendocrinology of childhood obesity. AB - The regulation of energy balance is enormously complex, with numerous genetic, hormonal, neural and behavioral, and societal influences. Although the current epidemic of obesity clearly has its underpinnings in the changes in culture during the past half-century (see other articles in this issue), the role of the neuroendocrine system in the genesis of obesity, as described in this article, is physiologically and therapeutically unavoidable. An understanding of this system has suggested organic causes (and therapies) for some rare and not-so-rare forms of obesity. With so many inputs, it is not far-fetched to assume that dysfunction of other parts of this feedback system will be found to explain other forms of obesity in the future. What does this mean for obese children entering the pediatrician's office? Fortunately or unfortunately, diet and exercise are the mainstays of obesity therapy for children and adults. Most diet-exercise programs result in an acute 11-kg weight loss in adults; the question is whether it can be sustained without significant long-term behavioral modification. For instance, the European Sibutramine Trial of Obesity Reduction and Maintenance trial showed that 42% of treated subjects drop out; of those remaining, 77% of subjects lost more than 5% of initial body weight, but only 43% of those maintained more than 80% of this over 2 years. Could there be an organic component in those who do not respond? Of course, obesity pharmacotherapies sometimes have beneficial acute effects, but these drugs work for only as long as they are consumed; discontinuation tends to result in a "rebound" weight gain, suggesting that the cause of the obesity is still present. Furthermore, in 2001, there are no obesity drugs approved for children. A useful guiding principle is that children deserve at the minimum an initial medical evaluation, including birth weight, medical history, family history, dietary evaluation, and exercise assessment. Perhaps the most important feature that can distinguish "organic" from "behavioral" weight gain in childhood is the age of the "adiposity rebound." The Centers for Disease Control and Prevention now supplies BMI charts for boys and girls at www.cdc.gov/growthcharts. Plotting of the BMI versus age allows pediatricians to determine the age at which the BMI starts to increase (mean, 5.5 years). The earlier the adiposity rebound, the more likely the child will be obese as an adult, and the more likely that an organic cause can be determined. In such patients, thyroid levels and fasting insulin and leptin levels should be measured. An initial attempt at diet and exercise is essential; patients who do not respond with BMI stabilization should be investigated for a more ominous cause of their obesity. As the nosology of obesity improves, pediatricians will be able to increase the diagnostic efficiency and therapeutic success of this unfortunate, debilitating, and expensive epidemic. PMID- 11494644 TI - Energy expenditure, physical activity, and obesity in children. AB - Although there are physiologic and genetic influences on the various components of energy metabolism and body weight regulation, and a major portion of individual differences in body weight can be explained by genetic differences, it seems unlikely that the increased global prevalence of obesity has been driven by a dramatic change in the gene pool. It is more likely and more reasonable that acute changes in behavior and environment have contributed to the rapid increase in obesity and that genetic factors may be important in the deferring individual susceptibilities to these changes. The most striking behavioral changes that have occurred have been an increased reliance on high-fat and energy-dense "fast foods," with larger portion sizes, coupled with an ever-increasing sedentary lifestyle. The more sedentary lifestyle is caused by an increased reliance on technology and labor-saving devices, which has reduced the need for physical exertion for everyday activities. Examples of energy-saving devices that have resulted in a secular decline in physical activity include: Increased use of automated transport rather than walking or biking Central heating and use of automated equipment, such as washing machines, in the household. Reduction in physical activity in the workplace because of computers, automated equipment, and electronic mail. Increased use of television and computers for entertainment and leisure activities. Use of elevators and escalators rather than stairs. Increased concern for crime, which has reduced the likelihood of outdoor playing. Poor urban planning that does not provide adequate biking paths or even sidewalks in some communities. Thus, the increasing prevalence, numerous health risks, and astounding economic costs of obesity clearly justify widespread efforts toward prevention efforts. These prevention efforts should begin in childhood because the behaviors are learned and continue through the lifetime. PMID- 11494645 TI - Dietary approaches to the treatment of the overweight pediatric patient. AB - A review of recently published studies on the dieting practices of children and adolescents examines the impact those practices are having on the overall health of youth who are overweight and those who are not. Recommendations for improving the nutritional quality of children's diets are made based on the food consumption patterns of youngsters ages 2 to 18 years. If implemented, these recommendations would lower calories, fat, saturated fat, and cholesterol, while improving nutrient density. Guidelines for the treatment of childhood overweight from four organizations are presented in chart form. PMID- 11494646 TI - Dietary fiber and body-weight regulation. Observations and mechanisms. AB - Dietary fiber may be related to body-weight regulation through plausible physiologic mechanisms that have considerable support in the scientific literature. Many short-term studies suggest that high-fiber foods induce greater satiation and satiety. Epidemiologic studies generally [figure: see text] support a role for fiber in body-weight regulation among free-living individuals consuming self-selected diets, although conclusive intervention studies addressing this point are lacking. Thus, there is considerable reason to conclude that fiber-rich diets, containing non-starchy vegetables, fruits, whole grains, legumes, and nuts, may be effective in the prevention and treatment of obesity in children. Such diets may have additional benefits, independent of changes in adiposity, in the prevention of cardiovascular disease and type 2 diabetes. PMID- 11494647 TI - Behavioral therapy in the treatment of pediatric obesity. AB - This article was designed to make a strong case for the importance of studying behavior and using behavioral therapy in the treatment of pediatric obesity. Behavioral treatments have been the most studied approaches to pediatric obesity, with great success. Six studies that provided long-term results are presented, and ideas for translating behavioral therapy into common pediatric practice are presented. Additional progress is needed to incorporate new findings in learning and behavioral neuroscience into clinical interventions and to integrate behavioral therapy with pharmacologic interventions and genetic predispositions and new advances in nutrition and exercise science. PMID- 11494648 TI - Exercise as a modality in the treatment of childhood obesity. AB - In a review of the literature, Glenny et al determined that family therapy and lifestyle modification seem to be effective in the prevention and treatment of childhood but not necessarily adult obesity. Furthermore, research indicates that obese children are better able to maintain weight loss over a long-term period than are adults. Based on the limited research in the treatment of obesity in children, approaches should include family interventions with nutrition and physical activity education, structured exercise, and behavior modification. These interventions should be delivered by a team of health care experts in a nurturing, nonintimidating environment; however, obese children respond differently physiologically and emotionally to exercise than do normal-weight children. Therefore, obese children may experience negative consequences to participation in activities considered appropriate for normal-weight children. In clinical settings, specialized exercise programs based on appropriate theories that include specific recommendations for children with varied obese conditions have been shown to enhance safety, efficacy, and compliance during pediatric obesity treatment. Optimal results may be achieved by combining programs to reduce sedentary behaviors based on similar theories with specialized, structured exercise prescriptions. When used in conjunction with appropriate dietary prescriptions and consistent behavior modification, exercise serves as a promising modality that may reverse obese conditions during childhood and, perhaps, prevent the onset of adult obesity. PMID- 11494649 TI - Exploring ways to reduce errors. PMID- 11494650 TI - Effects of orthostatic fluid shifts on bioelectrical impedance measurements. AB - This study analyzes the effect of orthostatic fluid shift caused by body postural changes on the reliability and stability of bioelectrical impedance. It has been found that following a fluid distribution perturbation caused by postural changes, impedance values change for a period of time that can be longer than the measurement time. Based on these preliminary measurements, it is recommended that new measurement protocols for bioelectrical impedance be developed in order to minimize perturbations on existing fluid distribution, thus increasing the reliability of bioelectrical impedance data. PMID- 11494651 TI - Evaluation of clinical thermometers for accuracy and reliability. AB - The purpose of this study was to examine the accuracy and reliability of a wide range of clinical thermometry instruments and technologies. In a historical sense, the purpose of this study was to determine if the improvements in speed, ease of use, and safety realized in the last 100 years have been offset by a loss of accuracy and/or reliability. In view of current events, the purpose was to determine if the new generation of electronic, digital clinical thermometers could be used to replace the traditional glass/mercury thermometers. Nine clinical thermometers representing electronic, digital oral, and predictive oral; electronic, digital infrared tympanic; and liquid crystal urinary technologies were evaluated. Accuracy was determined by comparing the temperatures obtained from these test instruments with those of the reference, glass/mercury oral thermometer. Reliability was determined by test-retest evaluation. All of the thermometers evaluated were significantly less accurate when compared with the reference thermometer in this study. All of the test instruments significantly underestimated higher temperatures and overestimated lower temperatures. This study indicated that the improvements in safety, speed, and ease of use of the newer clinical thermometers have been offset by a loss in accuracy and reliability. It also indicated that the current generation of electronic, digital clinical thermometers, in general, may not be sufficiently accurate or reliable to replace the traditional glass/mercury thermometers. PMID- 11494652 TI - How to create a capital process to maximize investments. PMID- 11494653 TI - Using UNIX--Part 1. PMID- 11494654 TI - [The effects of media violence on aggression: focus on the role of anger evoked by provocation]. AB - This study investigated the effects of anger evoked by earlier provocation on cognition, emotion, and aggressive behavior after being exposed to media violence. Sixty male undergraduates participated in the experiment. Before viewing one of three videos (either highly violent, violent with high entertainment, or nonviolent), half of the subjects were provoked by a confederate posing as another subject. Subjects' heart rates and eyeblink rates were recorded while viewing the video. After viewing the video, subjects described their thoughts that occurred while watching the video and rated their affective reactions toward the video. Finally, subjects' aggressive behavior toward the confederate was measured. Results of covariance structure analysis suggested that (a) anger evoked by provocation and high level of violence in videos additively elicited negative cognition and affect, which further facilitated aggressive behavior, and (b) high level of entertainment in videos elicited positive cognition and affect, which alleviated negative cognition and affect. PMID- 11494655 TI - [An examination of the relationships among career decision-making self-efficacy, vocational motives, and vocational indecision: a study of women's junior college students]. AB - The purpose of this study was to examine a causal model leading to the tendency of women's junior college students to delay vocational decisions. The model assumes that Career Decision-Making Self-Efficacy (CDMSE) determines vocational motives, which in turn affect the tendency. A questionnaire was administered to 431, 199 first-year and 232 second-year, women's junior college students. CDMSE was measured with self-efficacy for self-appraisal and occupational information gathering, and vocational motives were self-improvement, interpersonal, and status motives. Results showed that self-efficacy for self-appraisal influenced self-improvement motive for both first-year and second-year students. Self improvement motive and the self-efficacy then influenced vocational indecision among second-year students. Self-efficacy for information-gathering influenced vocational indecision among first-year students. These findings suggest that college vocational guidance should take self-efficacy and vocational motives into account. PMID- 11494656 TI - [The productivity of electronic brainstorming: a comparison of three systems with a control]. AB - Some studies have indicated that the brainstorming in a computer-mediated meeting can be effective. Accordingly, an experiment was conducted to investigate relative effectiveness of three electronic brainstorming systems, compared with a control, and find cognitive variables that mediated its effectiveness. One hundred undergraduate women, in groups of four, participated in the experiment. The number and quality of unique ideas generated by electronic brainstorming groups of three presentation systems: random, sequential, and sequence emphasized, were compared with those of a nominal group. Results indicated that the three brainstorming groups were higher than the control, in terms of originality of generated ideas. Also, the possibility was suggested that pleasure in task performance mediated the originality effect of electronic brainstorming. PMID- 11494657 TI - [Social exchange and inference: an experimental study with the Wason selection task]. AB - Social contract theory (Cosmides, 1989) posits that the human mind was equipped with inference faculty specialized for cheater detection. Cosmides (1989) conducted a series of experiments employing the Wason selection task to demonstrate that her social contract theory could account for the content effects reported in the literature. The purpose of this study was to investigate the possibility that the results were due to experimental artifacts. In the current experiment, the subject was given two versions of the Wason task that contained no social exchange context, but included an instruction implying him/her to look for something, together with the cassava root and the abstract versions used by Cosmides (1989). Results showed that the two versions with no social exchange context produced the same response pattern observed in the original study. It may be concluded that the subject's perception of the rule as a social contract was not necessary to obtain the original results, and that an instruction implying that he/she should look for something was sufficient. PMID- 11494658 TI - [Effects of figural complexity and target-distracter similarity on decay rate in short-term visual memory]. AB - We examined the effects of figural complexity and target-distracter similarity on decay rate in short-term visual memory (STVM) for closed contours. The subject's task was to compare two stimuli presented at different points in time. Experiment 1 showed that even a simple pattern with 6 convex parts cannot be handled in STVM for several seconds without loss. Experiments 1 and 2 indicated that decay rate tends to be higher for patterns with higher complexity and for patterns with higher target-distracter similarity regardless of pattern complexity. These results are interpreted that the value of each convex part represented in STVM is an independent Gaussian random variable and that memory noise increases with retention intervals for each convex part separately. Furthermore, they suggest that memory for local features in STVM decays faster. Yet visual rehearsal appears to operate effectively on memorizing even complex patterns. It is concluded that visually presented patterns can be retained in STVM with relatively high fidelity for prolonged periods of time, although some loss of precision occurs inevitably. PMID- 11494659 TI - [The effects of spatial information for cognitive distance in an architectural space]. AB - The present study investigated the effects of the spatial information for cognitive distance in an architectural space. One hundred undergraduate students participated in the experiment, and they were divided into two groups. The control group (CG) was given the pictures of both ends of the routes, and the experimental group (EG) was given some scenes of the routes in addition to the ends. The subjects have used the building used for the experiment for more than 2 years, and they estimated each walk of the 10 routes in the building. Each of those routes (50 m-144 m) is longer than the standard route (34 m). Before estimating the distance, the pictures were given on display through World Wide Web with questionnaires. The ratio method was used to obtain the cognitive distance data. As a result, the cognitive distance data of both groups showed similar distribution, and the cognitive distance data of EG were longer than those of CG in all routes. However, the effects of the number of the pictures were varied. These results suggested that not only the amount of the spatial information (i.e., chair, pillar, garbage box) but also the subjects' cognitive processing of the information affected the cognitive distance. PMID- 11494660 TI - [Young children's inference of moral disgust from situational and emotional response cues]. AB - The main purposes of this study were to examine young children's ability to infer disgust in another person elicited by someone else's immoral action, and their ability to properly use the person's reactions in order to infer his/her emotion. Stories with pictures were used, and these abilities were compared with those for anger, which was also elicited by someone else's immoral action. Children of five and six years old, 52 and 54 respectively, and 251 undergraduates participated in the study. Main results were as follows: (a) children were adequately able to infer disgust in another person from a story showing a situation with an immoral action, although not as well as undergraduates; (b) children used the person's reaction toward the immoral action for inferring disgust as well as undergraduates did; and (c) inferring disgust for them seemed to be as easy as inferring anger. PMID- 11494661 TI - [The relationship between flexibility of coping to stress and depression]. AB - The present study was conducted to examine the relationship between flexibility of coping to interpersonal stress and mental health as represented by scores on a self-report depression scale (CES-D) in 87 college students. Subjects were first required to complete the Interpersonal Stress-Coping Inventory (ISI) to identify each subject's coping type (ISIpre). They were then asked to complete ISI again (ISIpost) assuming that the coping strategy they adopted in ISIpre did not work well. The results of ISIpre and ISIpost were then compared and the flexibility of coping to stress of each individual was analyzed in terms of his/her tendencies: (a) to abandon the type they adopted in ISIpre (Level A flexibility); and (b) to adopt a new coping type (Level B flexibility). Flexible copers in both Levels A and B were shown to be significantly less depressive as represented by scores in CES-D, those fulfilling both flexibility criteria being the least depressive. The results were discussed with reference to mental health and to functional fixedness/recentering in problem solving. PMID- 11494662 TI - [Typicality effect on holistic processing in face recognition]. AB - Many studies have claimed that faces are more holistically recognized than other objects. It is unclear, however, whether some faces are more holistically recognized than other faces. This study examined whether typical faces are more holistically recognized than distinctive faces. In order to measure the degree of holistic processing, I used an alignment effect, which is a kind of interference effect on the part processing by the holistic processing. The alignment effect was measured as performance difference between aligned and non-aligned presentation conditions. The size of the alignment effects reflects the degree of the holistic processing. The results showed that the typical and distinctive faces showed an equal size of the alignment effects. These results suggested that the typical faces were not more holistically recognized than the distinctive faces. The implication of this results for face recognition research was discussed. PMID- 11494663 TI - Ridge preservation techniques using Bioplant HTR and Biofoil. Interview. PMID- 11494664 TI - [Antioxidant status of inhabitants of Yakutia]. AB - Presented are the results of antioxidant status of inhabitants of Yakutia. The intensity of lipid peroxydation depends on professional factors: influence of the quartz dust and smoking. Miners and smokers have high concentration of malonil dialdegide in blood. The SOD and catalase levels did not statistically depend on sex and nationality, but increased in miners and smokers. Insufficiency of vitamins C, A and E combined with immunoglobulins deviations. It should be noticed that country-people have more deviations in vitamin and immune status than towns-people. PMID- 11494665 TI - [Byproducts of grapes and carbohydrate metabolism]. AB - One of the important problems in treatment of diabetes is the preventing of considerable fluctuations of level of a glucose in a blood during day. In this connection it is recommended to reduce fat consumption, to prefer rations with a high content of dietary fibers. In experiences on rats is found, that a blood sedimentation rate, level of a hemoglobin did not differ from control magnitudes in conditions of long-lived trials of vegetable preparations. Amounts of reticulocytes, erythrocytes, lymphocytes, hematocrit were normal. An amount of thrombocytes and leukocytic formula practically did not change. Is established, that the glucose level in time gaps was reliably reduced when introduction of grape residues dietary fibers (5.9 +/- 0.7 mmol/l) and dietary fibers of grape seeds press cake (5.7 +/- 0.8 mmol/l) in a conventional ration as distinct from control (6.4 +/- 0.8 mmol/l). The low level content of dietary fibers in ration (5%) is reduce a level of a glucose on an empty stomach up to 6.8 +/- 1.4 mmol/l. The content of total protein in all groups of rats was in physiological norm and averaged 70-76 +/- 3 g/l. The investigated preparations of grape residues dietary fibers and dietary fibers of grape seeds press cake are nontoxic, are well tolerated laboratory animal, and can be recommended as the treatment and prophylactic components for correction of carbohydrate and lipide dysbolism. PMID- 11494666 TI - [Use of multivitamin drink and oil solution of beta carotene in the treatment of children with chronic gastroduodenitis]. AB - Inclusion of vitamin-containing soft drink and oil solution of beta-carotene in the diet of the 4-14 years old children suffering from gastrointestinal diseases lead to vitamin and antioxidant status improvement with simultaneous positive effects on their clinical condition. Polyhypovitaminosis frequency had decreased. Thus, there is every reason to fortify diets of the patients suffering from gastrointestinal diseases with vitamins-antioxidants. PMID- 11494667 TI - [Correction of mineral metabolism impairment with an allowance for seasonal fluctuations in patients with ischemic heart disease and hypertensive disease with overweight]. AB - The influence of antiaterosclerotic diet with including some biologically active supplements (BAS), containing Zn, Mg, Na, K, Ca, vit C, E, B2, B6, beta-carotene, I was studied in 164 patients with ischemic heart disease and hypertension disease. BAS has been use with an allowance for seasonal fluctuations of microelements sufficiency. The usage of BAS "Lifepack" has priority in spring. The usage of biologically active supplements during 4 weeks in spring has promoted positive changes of clinical symptoms of diseases against a background of lowering increasing of K, Ca, Mg, Cu, Zn. PMID- 11494668 TI - [Comparative study of effects of diet therapy including eiconol or linseed oil on several parameters of lipid metabolism in patients with type 2 diabetes mellitus]. AB - It was investigated the influence of a diet supplemented with polyunsaturated fatty acids (PUFA) omega-3 on dynamic of basal glycaemia, lipid profile and composition of fatty acids of erythrocytes in type 2 diabetic patients. Traditional 9 diet was supplemented with fish ("Eiconol", 8 grams per day) or vegetable (linseed oil, 17.5 grams per day) PUFA omega-3. The results investigations indicated that a omega-3 PUFA-enriched diet has a beneficial effects on lipid metabolism. But dietary administration of PUFA omega-3 may be used differently for correction of dislipoproteinemia in diabetic patients. PMID- 11494669 TI - [Hygiene and standards aspects of registration, marking and labeling of food products prepared from genetically modified sources]. AB - The basic requirements to problems of registration, marking and labelling of the foodstuff prepared from generically modified sources are stated. The comparative analysis of the above-named problems in the different countries (USA, countries of EU, Russia etc.) is given. PMID- 11494670 TI - [Toxicological and hygienic assessment of nitrates in food products]. AB - The toxicological and hygienic estimation of the contents of nitrates in the basic vegetable cultures (beet, carrots, potato etc.) is given. Is shown, that nitrates, contained in the increased amounts in the aforesaid cultures are not indifferent for human and animals organism. At a unbalanced diet of cattle use of forages with the increased contents of nitrates can result in occurrence them in livestock products. The hygienic regulation of permissible concentrations nitrates in some products should be carried out with an allowance fo structure of nutrition of the population, contents nitrates in drinking water, and also other factors capable to influence on formation of nitrate body burden on the population and to aggravate effect of nitrates action. PMID- 11494671 TI - [Method of identification of natural vegetable oils]. AB - The natural compounds which are present in vegetable oils were investigated by absorption and three-dimensional fluorescence spectroscopy. It was found that some of these components could be used as internal markers for identification and quantitative analysis of vegetable oils. The presence of specific fluorophores permits to develop an individual fluorescent "finger-print" criteria for all the studied oils, which can be used for detection of product falsification. PMID- 11494672 TI - [Thermodynamic approach to the selection of polyuronide sequestrants for protection of the human body from toxic metal ions. Interactions of polyuronides with lead ions]. AB - Binding isotherms of Pb2+ ions with potassium pectate and potassium alginate with relatively low content of blocks of L-guluronic residues (20%) have been determined. Interactions of Pb2+ ions with polyuronides studied is cooperative. Maximum values of binding constants are an order of magnitude higher than previously determined ones for Ca2+ and Sr2+ ions. Along with ion-coordination ("stoichiometric") interactions, alginate is typified by so-called extra stoichiometric binding of Pb2+ ions, which presumably proceeds by a coprecipitation mechanism. Limitations of the thermodynamic approach to the selection of sequestrants for human body protection from toxic metal ions are discussed. PMID- 11494673 TI - [Role of educational and scientific programs in realization of the concept of healthy nutrition in Siberia]. PMID- 11494674 TI - [Dynamic examination of the character of school children's nutrition in Kabardino Balkaria]. AB - The results of dynamic examination 842 high school students 10-17 years old, boys and girls, in the period of socioeconomic conversions in 1992-1998 are presented. An actual nutrition was studied by a reference method of diurnal inquiry. Expressed disbalancement in nutrition consisting in increased consumption of saturated fatty acids (13.3%), saccharum (15.8%) on a background of the decreased consumption of total carbohydrates (52.8%), oligosaccharides (6.2%), polyunsaturated fatty acids (4.5%), animal protein (6.7%), cellulose (2.6 g) and practically of all micronutrients is found. The low content in rations a fish, fruit, vegetables, nuts, dairy produce and high--confectionery, baked products, drinks is shown. Reduction of energy value of daily rations and their contents of series of alimentary substances have aggravated disbalancement of children's nutrition. PMID- 11494675 TI - Possibility of paternity testing using RFLP analysis on a very small amount of material. AB - Paternity testing by DNA analysis was carried out using dental pulpal and chorionic villous tissue from two children respectively, and fresh blood samples obtained from the alleged parents. DNA was extracted spectroscopically from the pulp of an upper wisdom tooth (16 micrograms) and the chorionic villi (53 micrograms). The RFLP method was used for DNA analysis of the parent-child relationships because both of the DNAs extracted had a high molecular weight. Distinct bands were detected with 32P-labelled multi-locus (Myo) and single locus (pYNH24) DNA probes. In the case of the dental specimen all of the bands of the child's DNA were found to be derived from either of the alleged parents, demonstrating a consistent parent-child relationship (the probability of established paternity was 99.86%) whilst in the case of the villous specimen the father-child relationship was denied. This procedure can provide much information using very little material for analysis but where the samples are in a good condition. PMID- 11494676 TI - Diet and age-at-death determinations from molar attrition. A review related to the low countries. AB - To elucidate the impact of diet on age-at-death determinations based on molar attrition a comparison was made between the established rate of attrition in three populations; a pre-mediaeval (British), a late mediaeval (Dutch) and a 17 18th century (Dutch) (western European). It appeared that the rate decreased dramatically during the overall time span and that this change was probably diet related and owing to the coarseness of foodstuffs. This result strongly indicated that molar age-attrition tables should only be used for age-at-death determinations if their application is restricted to a particular cultural period and diet. PMID- 11494677 TI - Identification from a bitemark in a wad of chewing gum. AB - A wad of used chewing gum recovered from the scene of a burglary contained impressions of human teeth. Casts of these impressions displayed unique morphological characteristics which were found to show concordance with corresponding features present on casts of the posterior teeth of a suspect. PMID- 11494678 TI - A review of the most commonly used dental age estimation techniques. AB - This review of literature provides an overview of the most commonly used dental age estimation techniques and focuses on dental age estimation scoring systems in children and adults. In order to obtain a more reliable and reproducible age estimation the forensic odontologist should use several of these available methods whenever an age estimation in the living or dead is required. PMID- 11494679 TI - Pre-ortho mandibular joint assessment. PMID- 11494681 TI - Intra-oral wire bending for tip edge treatment. PMID- 11494680 TI - The Chin-Button. PMID- 11494682 TI - TMD: after the orthotic. PMID- 11494683 TI - Segmental archwire mechanics. AB - Straight wire orthodontic methods cannot treat the significant compromises demonstrated in this article. These types of patients require a three-dimensional method of correction, which involves segmental orthodontics or a modified straight wire technique. This article will discuss how to use the information gathered from a three-dimensional evaluation of the stable mandibular/maxillary relationship using a diagnostic orthotic to treat to a three-dimensional balanced relationship. Structural and functional relationships exist between the final occlusal relationship and the function of the joints, muscles and upper quadrant that must be considered during orthodontic, orthopedic and restorative finalization procedures if long term permanence is to be obtained. This article demonstrates these relationships and provides a means to approximate the dentition to function in harmony with the associated structures. PMID- 11494684 TI - [Delayed birth of the second child in multiple gestation]. AB - Five case histories illustrate the issue of delayed interval deliveries. In the first two cases, the first child was born at a gestational age of 20 and 18 weeks, respectively. The first woman (40 years old) gave birth to the second child after successful prolongation of pregnancy to a gestational age of 38 weeks. In the second case (28 years old), the attempt to delay delivery failed and the second child was born at 19 weeks of gestation. The third case (32 years old), illustrates the enormous differences in neonatal course between a child born at 26 weeks of gestation, who had to be treated at length for respiratory distress syndrome, hypotension and patent ductus arteriosus, and his twin brother born two weeks later and who recovered more quickly. The fourth case (24 years old) describes delayed delivery to allow administration of antenatal glucocorticoids. The last case (32 years old) deals with a serious maternal complication of placental abruption during an attempt to delay the birth of the second twin. Early tocolytic and antibiotic therapy may delay delivery and, in combination with antenatal glucocorticoids to stimulate lung maturation, may thereby improve the condition of the second twin. The role of cervical cerclage remains controversial. There is an important publication bias in the literature due to under-reporting of the failed attempts of delayed deliveries. In multiple gestation with imminent very preterm birth, delayed delivery of the second child is a feasible management option. PMID- 11494685 TI - [Erectile disorders: the reality]. AB - The grouping together of erectile dysfunction and erectile disorder (i.e. dysfunction with distress) has led to the presentation of incredibly high prevalences (up to 52%). When limited to erectile disorder, two Dutch open population studies, among men aged 40-79 years, show remarkably similar and more realistic prevalences, namely 3 to 10%. Although the 'Leiden Impotence Screening Test' appears to reliably exclude somatic aetiological factors, it would be preferable if the general physician posed such diagnostic questions. It is hoped that the availability of pharmacotherapeutic treatments (notably sildenafil) will not tempt the general physician to join the patient in his inclination to 'somatise' his erectile disorder. Both intracavernosal self-injection therapy and the implantation of an erectile prosthesis give rise to a large proportion of dissatisfied men. In the Netherlands to date, the number of continuation prescriptions for sildenafil equals the number of first prescriptions, which suggests that many men also stop with this therapy. In the Netherlands, health insurance companies only reimburse self-injections and prosthesis implantations. It can be argued that all cost-effective treatments for erectile disorder, including psychosexual therapy, should be reimbursed, or none at all. PMID- 11494686 TI - [Report of the Dutch Health Council on the cost-effectiveness of long-term psychotherapy]. AB - A recent report from the Health Council of the Netherlands considers the cost effectiveness of long-term psychotherapy (more than 25 sessions over a period of more than one year). It is concluded that knowledge about the efficacy and effectiveness of this form of therapy is scarce, and the cost-effectiveness is largely unknown. A plea is made for further research and for restraints in the administration of such therapy. However, research into the efficacy of long-term psychotherapy requires a controlled design, with the selection and randomisation of patients. This precludes definitive conclusions with regard to the cost effectiveness in open patient groups. Moreover, psychotherapy is not only aimed at a cure, but also to a fairly large extent at care. Finally, if it is difficult to determine the efficacy of psychotherapy then this is even more so the case for long-term psychotherapy, as the success may be attributable to a wide range of intervening events that occur during the course of treatment. PMID- 11494687 TI - [From limitations to possibilities; the advice from the Donner Committee about disability benefits legislation]. AB - In the Netherlands, the number of people declared unfit to work continues to rise: every working day sees more than 100 new benefit claimants. The average age of these claimants is decreasing and the number of women declared unfit for work is showing a disproportionate increase. Against this background, the Dutch government appointed a committee in June 2000 whose task was to examine the functioning of the legislation. The Donner Committee concluded that the claims made under the disability benefits legislation could not be clarified by a worsening of either the public health or the working conditions. The legislation is based on obsolete ideas in which illness effectively excludes employment. This idea fosters a range of social problems in the employment process, which present themselves as medical problems and therefore remain elusive and unsolved. The result is that people are unnecessarily eliminated from society at a time when paid employment is an increasingly important means of social interaction. The Committee proposes a drastic simplification of the legislation so that a disability benefit can only be claimed in the case of complete unfitness for work. Such a claim can only be made in the case of a serious illness which results in permanent and considerable limitations in everyday functioning. For all other cases involving health problems, the employer and employee need to work together to organise appropriate work. The emphasis is on what the employee can do as opposed to what he/she cannot do. If the employer fails to take the necessary measures then he/she will be obliged to continue paying the salary and if the employee fails to cooperate then he/she can be dismissed. If needs be the employee must be prepared to except a salary reduction of 30%. After two years of absence, dismissal is in every case possible. It is anticipated that as a result of the Committee's proposals, more people will remain in employment but that the number of unemployed as well as the number of legal disputes concerning dismissal will increase. PMID- 11494688 TI - [Age-related macular degeneration; therapeutic options]. AB - The prevalence of age-related macular degeneration varies from 1% in patients aged 65 to 74 years, to 10% in patients aged 85 years and older. In the Netherlands, there is now a total of around 55,000 patients with age-related macular degeneration, and this figure is expected to rise to approximately 114,000 by 2025. Loss of central vision is caused by a degeneration of photoreceptors cells and pigment epithelium. Age-related macular degeneration is subdivided into a dry, atrophic form and an exudative form with neovascularisations. Many international studies have been published over the past ten years on potential new therapies for neovascular macular degeneration. For conventional laser treatment, it has been shown that visual loss in the long term is less marked in the treated group than in the observation group, despite the immediate destruction of the retina. Photodynamic therapy seems a promising new form of local therapy with few side-effects. For both therapies, however, the indication is limited to patients with the classic type of neovascularisation. Data on radiotherapy, submacular surgery and macular translocations are not sufficiently well-founded for firm conclusions to be drawn. PMID- 11494689 TI - [Gastrointestinal surgery and gastroenterology. XIII. Classification and diagnosis of perianal fistulas]. AB - Intersphincteric abscesses due to inflammation of an anal gland are the origin of most perianal fistulas. Perianal fistulas are classified into four types. Intersphincteric and transsphincteric fistulas are the most common. The appropriate surgical treatment of perianal fistulas is dependent upon their correct classification. Despite new imaging-techniques, physical examination of the perianal region remains of paramount importance for the preoperative classification of perianal fistulas. At present, endoanal ultrasound with the injection of hydrogen peroxide into the fistulous tract, is advocated as the most suitable method for the preoperative imaging of perianal fistulas. PMID- 11494690 TI - [Diagnostic image (47). Zenker's diverticulum; diverticuli in the small intestine; Meckel's diverticulum]. AB - Zenker's diverticulum; diverticula of the small intestine; Meckel's diverticulum. PMID- 11494691 TI - [Low 5-year recurrence rate after surgical excision of 126 basal cell carcinomas with frozen section analysis upon indication]. AB - OBJECTIVE: To determine the recurrence rate of basal cell carcinoma (BCC) after surgical excision. DESIGN: Retrospective. METHOD: The records of all 139 patients treated for BCC at the Department of Plastic Surgery and Hand Surgery, Leeuwarden Medical Centre, Leeuwarden, the Netherlands, in 1992 were reviewed. This was done by examining case histories and additional information was obtained from dermatologists, general practitioners, patients and the Dutch National Automated Pathological-Anatomical Archive. One hundred and twenty-six BCCs were investigated in 108 patients: 51 men and 57 women, with a mean age of 66 years (range: 24-92). Of these BCCs, 114 had not been previously treated and 12 were a relapse. Freeze section analysis was carried out during the excision if a sprouting or deep growth was suspected and if a tight excision or reconstruction with transposed or rotated piece was desired. RESULTS: The mean follow-up was 59 months (range: 5-86). Sixty-one BCCs on which frozen section analysis was performed were excised completely. In 65 BCCs which were excised without frozen section analysis, a second operation was necessary on 11 occasions to obtain histologically free margins. In both groups a tumour recurred on one occasion (2/126; 1.6%). CONCLUSION: Surgical treatment of BCC aimed at free histological margins, results in a recurrence rate of 1.6%. PMID- 11494692 TI - [Labile hypertension due to iatrogenic denervation of the carotid sinus]. AB - Two patients, a man and a woman aged 36 and 40 years, respectively, were found to have disturbed blood pressure regulation after medical intervention in the jugular area. The man had undergone radiation therapy of the neck due to regional lymph node involvement of a nasopharyngeal carcinoma. Six years later he developed episodic complaints of headache accompanied by labile hypo- and hypertension. Mental and physical stimuli resulted in excessive sympathetic activation. In the woman, a carotid body tumour was resected bilaterally. There was an immediate onset of hypertension after surgery. In both patients, the clinical syndrome was attributed to afferent baroreflex failure due to carotid sinus denervation. PMID- 11494693 TI - [Specialists and assistant physicians; disciplinary division of responsibilities]. AB - In every day hospital practice, the division of responsibility between specialists and assistant physicians plays an important role. This division of responsibility has repercussions in disciplinary judgements concerning the conduct of the supervisor and the assistant physician. The problem with this is that disciplinary law assumes individual liability. If a patient chooses to lodge a claim solely against the assistant physician then whether the assistant physician can appeal to shortcomings in the supervision is dependent upon the situation concerned. The supervisor is obliged to ensure that the necessary supervision is given or if the assistant physician indicates that a given task is beyond his own abilities then the supervisor should carry this out or find someone else to carry it out on his behalf. Within jurisprudence the value of protocols is becoming increasingly important. PMID- 11494694 TI - [Teaching to inhale; better utilization of powder inhalators after counseling and instruction]. PMID- 11494696 TI - [Roaming through methodology. XXXII. False test results]. PMID- 11494695 TI - [Pharmacotherapy for neuropathic pain caused by injury to the afferent nerve fibers]. PMID- 11494697 TI - [Roaming through methodology. XXXII. False test results]. PMID- 11494698 TI - [Little health benefit gained with hormone replacement therapy in postmenopausal women in the Netherlands]. PMID- 11494699 TI - [Enterobacter cloacae epidemic in a neonatal intensive care unit due to the use of contaminated thermometers]. PMID- 11494700 TI - [CBO guideline 'Larynx carcinoma']. PMID- 11494701 TI - [Pain in the symphyseal region after parturition; possibly osteomyelitis]. PMID- 11494702 TI - Policy, politics, postures, and programs. PMID- 11494703 TI - Survey finds specialists more likely to recommend appropriate treatment. PMID- 11494704 TI - "Punctuated" HIV meds planned for TB patients. PMID- 11494705 TI - Recent advances in the management and treatment of GI and hepatic diseases associated with HIV: Part I. PMID- 11494706 TI - NIH launches online AIDS oral history project. PMID- 11494707 TI - New medical facility will train health providers from all of Africa. PMID- 11494708 TI - HIV treatment adherence research. PMID- 11494709 TI - The changing face of mycoses in patients with HIV/AIDS. AB - The current era of effective antiretroviral therapy has led to a marked reduction in opportunistic infections (OIs) in those countries where such therapies are available. Opportunistic fungal infections are no exception, and the incidence of such infections is now 20% to 25% of that seen in the mid-1990s. Infections associated with very advanced HIV disease, such as azole-resistant candidiasis and aspergillosis, are also rarely seen, reflecting the improvement in immune function. Indeed, the most common issue now is whether patients who have had a systemic mycosis require life-long therapy as had been recommended. Preliminary data from small studies suggest that as with other OIs, it may be possible to stop suppressive therapy in patients with a history of mycosis whose CD4+ lymphocyte count rises with antiretroviral therapy. Thus, it appears that the future of HIV-associated mycoses is linked to the future of effective treatment for HIV itself. PMID- 11494710 TI - Nevirapine-induced hepatitis: a case series and review of the literature. AB - Nevirapine, a nonnucleoside reverse transcriptase inhibitor used as part of combination antiretroviral therapy, can cause mild elevations in transaminase levels. Severe elevations in transaminase levels related to the use of nevirapine developed in 4 patients. Data on these patients were extracted via chart review, and a review of the literature was also completed. Nevirapine-induced hepatitis occurred shortly after drug initiation in patients with and without preexisting liver disease. Significant elevations in liver enzyme levels occurred but resolved promptly in most with discontinuation of the nevirapine. Close monitoring of liver enzyme levels in the early period after starting nevirapine is essential. PMID- 11494711 TI - [Cajal in current urology]. PMID- 11494712 TI - [Urologic disease emergency. Clinico-epidemiologic analysis at a district hospital]. AB - OBJECTIVE: To present the results of a one-year retrospective study that analyzed the clinical and epidemiological aspects of urological emergencies, which is not a negligible percentage of all hospital emergencies. METHODS: Data on all the urological emergencies of the emergency department of our hospital were collected during a one-year period by filling out a form that included sex, age, whether referred or not, disease, etc. RESULTS: There were 1504 urological emergencies; the urologist was required in 458 of these cases (30.45%). There were more men (837) than women (667). Patient ages ranged from 1-101 years (mean 53 years); a higher incidence was found for those aged 65 years. By age groups, there were more patients aged 61-80 years (575 patients). Most of the patients came to the hospital on their own initiative (1114) and 390 had been referred by their primary care physician. Renal colic (670 cases) was the most common diagnosis in both males and females, followed by cystitis, which was prevalent in the females (67.35%). Other conditions seen were urinary retention, hematuria, etc. CONCLUSIONS: There was a higher prevalence of males that consulted for urological conditions at the emergency department and there was a higher incidence in patients in the 6th and 7th decades of life, which is similar to the findings reported in other studies. Renal colic was the main reason for consultation for both genders and for all age groups, except those younger than 20 who presented more scrotal and penile involvement. Infective conditions involving the lower urinary tract (cystitis) and pyelonephritis were prevalent in the females. The large number of patients that were discharged from the emergency department and who consulted on their own initiative shows their confidence in the emergency services and that they could have been attended to at other health centers and did not have to come and unnecessarily strain the hospital emergency services. PMID- 11494713 TI - [Putative chromophobe cell renal carcinoma: are they or are they not?]. AB - OBJECTIVE: According to the literature, the typical histological findings and simple colloidal iron staining permit the identification of chromophobe cell renal carcinoma, a genetically well-established entity. Our doubts on whether this tumor type can be recognized by conventional methods are presented in this study. METHODS: 130 cases of renal carcinoma were treated from 1977 to 1997. Of these, 12 showed characteristic general histological features compatible with chromophobe cell renal carcinoma and reticulated and intense, diffuse cytoplasmic positivity on colloidal iron staining. These tumors were reviewed for the following: 1) gross appearance, 2) architecture, 3) cytoplasmic characteristics, 4) nuclear characteristics, 5) colloidal iron histochemical staining which is considered fundamental and exclusive, 6) immunohistochemical phenotype. Ultrastructural study of material fixed in paraffin was also performed. RESULTS: One case met all 6 criteria, 3 met 5 of them, 6 cases met 4, and 2 cases met 3 of the criteria. The ultrastructural study was not useful in making the diagnosis due to the poor quality of the material. CONCLUSIONS: If all cases were chromophobe cell renal carcinoma, then this tumor type can be recognized and diagnosed by simple techniques in any pathology laboratory, and its incidence, presence of necrosis, hemorrhage and high nuclear grade would be even greater than currently accepted. Furthermore, it would be expected that the prognosis, by each grade and stage, would not be so different from that of the conventional renal carcinoma, as some large series have already indicated. On the other hand, if our cases or some of them were in fact conventional renal cell carcinoma that closely resembled chromophobe cell renal carcinoma, an exact diagnosis cannot be made without a genetic or ultrastructural study (using adequately fixed material), and most of the published studies would therefore have to be questioned. PMID- 11494714 TI - [Renal cancer: descriptive analysis of a series of 267 intervened cases]. AB - OBJECTIVE: To present the results of a descriptive analysis of 267 surgically treated renal tumors. METHODS: From January 1986 to October 1999, 267 patients (153 males and 114 females) with renal tumor were treated in our department. All data were introduced into the Access data base program and analyzed using the SPSS software. Descriptive analysis was performed and life expectancy was calculated with the Kaplan-Meier survival curve. Mean follow-up was 42.72 months. RESULTS: 56.4% were in the right and 43.6% were in the left kidney. 41.2% of the cases were incidentally discovered. In the symptomatic patients, the most common presenting feature was hematuria (51.8%). Radical nephrectomy was performed in 94.2%, partial in 3.1% and tumor resection in 2.7%. The lumbar approach was used in 75.5% of the cases. In regard to the histology, 88.7% were carcinomas. Clear cell was the most frequent cell type (91.5%). By grade, 55.8% were GI, 32.9% GII and 11.3% were GIII. By stage (according to the 1992 TNM classification), 64.3% were stage I, 15.4% stage II, 17% stage III and 3.3% stage IV. 80.5% showed no vascular involvement. The 5-year overall survival was 71.93% (mean 114 months; median 167 months). Currently, 76% of the patients are alive. CONCLUSIONS: Distribution by sex in our series was different to that reported in most of the studies. The tumor was incidentally discovered in a high proportion of the cases. The lumbar access was the most widely used surgical approach. The overall survival is similar to that reported by other groups. PMID- 11494715 TI - [Trigonocervicoprostatotomy. Results in 205 cases]. AB - OBJECTIVE: To analyze the results of transurethral incision of the prostate in 205 patients with prostatic obstruction. METHODS: 205 patients comprised the study. The clinical records and surgical reports were reviewed for patient age, pre and postoperative symptoms, physical examination (principally digital rectal examination), ultrasound findings (prostate weight), endoscopy, type of surgery (type of incision), operating time, hospital stay and complications. RESULTS: 37.07% were aged 60-69 years, 59.51% were grade I according to the DRE and ultrasound findings, and prostatic weight. Bilateral incision was performed in 65.36%. The operating time was 15 minutes in 79.02% of the cases and the length of stay was 3 days in 89.75% of the cases. Postoperative hematuria was the most common complication. CONCLUSIONS: Endoscopic transurethral incision of the prostate is an alternative minimal invasive surgery in the treatment of early onset benign hyperplasia of the prostate with small prostates and short urethras. PMID- 11494716 TI - [Nephrotic syndrome and anasarca status, secondary to treatment with tiopronin in a case of cystinuria]. AB - OBJECTIVE: To describe a case of cystinuria treated with tiopronin that produced the nephrotic syndrome. METHODS/RESULTS: A case of nephrotic syndrome with ascites and heart failure in a patient who had received tiopronin for the treatment of cystinuria is presented. Cystinuria as a rare cause of kidney stones is analyzed. The clinical features, diagnosis and the side effects of treatment with tiopronin are discussed. The patient recovered after withdrawal of the drug. CONCLUSIONS: It must be emphasized that patients treated with tiopronin should be screened for proteinuria. PMID- 11494717 TI - [Diverticuli of the female urethra. Diagnosis with transrectal ultrasonography]. AB - OBJECTIVE: To analyze the role of transrectal ultrasound in the diagnosis of diverticula of the female urethra. METHODS/RESULTS: Transrectal ultrasound assessment was performed with a biplanar 5 MHz probe and translabial ultrasound was performed with a 7 MHz linear or 3.5 MHz sectorial transducer in 9 women suspected to have urethral diverticula. Cystourethrography was also performed in three patients. Eleven diverticula were found in these 9 patients. CONCLUSIONS: The incidence of diverticula of the female urethra has been reported to range from 0.5-6%. Diagnosis is confirmed by urethroscopy or imaging methods. Transrectal ultrasound is the method of choice in the assessment of urethral diverticulum. PMID- 11494718 TI - [Abscess of seminal vesicle: conservative treatment]. AB - OBJECTIVE: To describe a case of primary seminal vesicle abscess that was managed conservatively. METHODS: Antibiotics, according to the urine culture and antibiotic profile, were administered for four weeks. Outpatient follow-up was carried out with digital rectal examination, urine culture and ultrasound. RESULTS: A favorable clinical response was observed at day three. The patient was discharged from hospital on day 7. DRE, urine culture and ultrasound findings were normal after 4 weeks. CONCLUSIONS: Primary abscess of the seminal vesicle can be successfully treated with antibiotics. Percutaneous drainage of the abscess should be reserved for those cases that do not respond to conservative management with antibiotics. PMID- 11494719 TI - [Adrenal ectopy in epididymis]. AB - OBJECTIVE: To report a case of ectopic adrenal in the epididymis. METHODS: A 17 year-old male consulted for a left varicocele and complaints referred to the right epididymis a year and a half after an episode of acute epididymitis. A nodule was excised from the epididymis. RESULTS: The pathological analysis showed an 0.4 cm nodule comprised of adrenocortical tissue. CONCLUSIONS: Ectopic adrenal cortical tissue is a benign lesion. Although routine search for this lesion is not indicated, it should be resected for histological analysis and differential diagnosis when it is found during a surgical procedure in this area. PMID- 11494720 TI - [Crossed renal ectopy with fusion: report of a case]. AB - OBJECTIVE: To present a case of crossed fused renal ectopia that was incidentally discovered in a patient consulting for abdominal pain in the emergency department. METHODS: A 46-year-old patient consulted at the emergency department for abdominal pain. Assessment by diagnostic imaging techniques demonstrated a crossed renal ectopia and associated bone anomalies. RESULTS/CONCLUSIONS: Crossed renal ectopia is an uncommon congenital anomaly and in most of the cases usually presents with fusion of both kidneys. It can also be associated with congenital anomalies of other organs. No treatment is required unless there are other complications or superimposed pathologies. PMID- 11494721 TI - [Extrinsic ureteral obstruction secondary to appendiceal mucocele]. AB - OBJECTIVE: To report a case of extrinsic ureteral obstruction arising from mucocele of the appendix and review the clinical, diagnostic and therapeutic aspects of this appendiceal pathology of interest in urology. METHODS: A 71-year old female patient that had previously undergone radical gynecological surgery and postoperative radiotherapy presented with right lumbar pain. Patient evaluation by ultrasound, IVP and CT showed a pelvic mass causing right extrinsic ureteral obstruction. Release and reinsertion of the right ureter and appendectomy were performed. RESULTS: The anatomopathological study showed an appendiceal mucinous cystadenoma. The patient is asymptomatic after surgery. CONCLUSIONS: Appendiceal mucocele is a dilatation of the vermiform appendix due to collection of mucoid material that is frequently produced by a cystadenoma. The presenting feature is usually that of acute appendicitis. The definitive diagnosis is based on the anatomopathological findings. Treatment is by appendectomy for the benign tumors and by right hemicolectomy in a second stage procedure for malignant tumors. This appendiceal pathology should be taken into account in the differential diagnosis of right extrinsic ureteral obstruction. PMID- 11494722 TI - [Vesico-scrotal hernia with hypocontractile detrusor]. AB - OBJECTIVE: To present a case of massive inguinoscrotal hernia associated with low detrusor contractility. METHODS/RESULTS: A 70-year-old male patient with a history of diabetes and a previous diagnosis of low contractile bladder presented with urinary retention. Acute renal failure secondary to ureteral obstruction and inguinal bladder herniation was diagnosed. The direct bilateral hernia was repaired. Postoperative urodynamic evaluation showed an acontractile bladder. Intermittent catheterization was started, with satisfactory functional result and improvement of renal function. CONCLUSIONS: Massive bladder hernia can present as acute renal failure without lower urinary tract obstruction. The underlying impaired detrusor contractility can be diagnosed by urodynamic evaluation. Surgical treatment, with self-catheterization if required, achieves good results with preservation of renal function. PMID- 11494723 TI - [Adrenocortical macronodular hyperplasia with massive enlargement of the gland. A rare variant of autonomous hypercortisolism]. AB - OBJECTIVE: To present a case of Cushing's syndrome due to massive adrenocortical macronodular hyperplasia that gave the gland the appearance of a pseudotumor. METHODS: In a review of the surgical pathology of the adrenal gland recently performed in our hospital, 82 adrenalectomy specimens obtained from 1978-1998 were found. Of these 82 surgical specimens, 44 (53.6%) were cortical pathologies and 10 of these (12.1%) were hyperplasia. There was only one with massive adrenocortical macronodular hyperplasia. A 53-year-old male, chronic alcoholic with cushingoid obesity and hypercortisolism is described. RESULTS: Ultrasound and CT assessment showed enlarged, asymmetrical adrenal glands and macronodules. The analytical and MRI study of the sella turcica showed primary adrenal hypercortisolism. Adrenalectomy of the dominant gland (the left adrenal whose activity was demonstrated on the 131Iodine scintiscan) was performed. The postoperative cortisol levels returned to normal and remain normal up to the present time, 18 months after surgery. The surgical specimen weighed 104 gms and was completely macronodular. CONCLUSIONS: Massive adrenocortical macronodular hyperplasia is a rare cause of pituitary-independent autonomous primary adrenal hypercortisolism. It is always bilateral. Adrenal weight is significantly increased (total weight of both adrenals 60-180 gms) and present yellow or golden, non-encapsulated nodules of a few millimeters to up to 4 cms, comprised of cells with scanty activity, therefore massive enlargement of the glands is necessary to produce Cushing's syndrome. The clinical, biochemical and surgical aspects can cause confusion and can be suggestive of an adrenocortical neoplasm. Treatment is by adrenalectomy. PMID- 11494724 TI - [Primary adenocarcinoma of the ureter]. AB - OBJECTIVE: To discuss the diagnosis and treatment of primary adenocarcinoma of the ureter with presentation of a case. METHODS/RESULTS: A female patient presented with renal colic. Histological analysis showed a primary adenocarcinoma of the ureter. Segmental ureterectomy was performed with good results. CONCLUSIONS: Adenocarcinoma of the ureter is not very frequent. Diagnosis is usually based on the histological findings. Segmental ureterectomy can be a surgical alternative. PMID- 11494726 TI - Intraoperative sentinel node biopsy in patients with squamous cell carcinomas of the head and neck--experiences using a well-type NaI detector for gamma ray spectroscopy. AB - The sentinel node (SN) biopsy in an actually discussed topic. The use of a well type NaI detector for gamma ray spectroscopy (WTD) beside the use of a handheld gamma probe (HGP), is investigated. In 18 patients suffering from a head and neck squamous cell carcinoma (HNSCC) with different lymph node status an intraoperative SN biopsy was accomplished with a HGP. 64 separately taken lymph nodes were supplied to an additional measurement in a WTD. A tumor-free SN represented the tumor-free lymph node status in 9 patients. In 5 cases an isolated tumor metastasis could be proven in the SN. SN biopsy has no meaningfullness in cases of advanced metastatic spread ipsilaterally, but possibly in contralateral N0-neck. In 64 separately investigated lymph nodes an activity enrichment could be proven in the WTD. With this procedure small count rates could be determined in contrast to the HGP. The distinction of weakly enriching lymph nodes was less favourable with the HGP but successful with the WTD. The additional use of a WTD offers a more exact distinction of intranodal disintegration rates of the draining lymph nodes. The WTD may increase the security of intraoperative SN biopsy. PMID- 11494725 TI - The TURP syndrome: importance of expiratory ethanol measurement and high serum levels of glycine. AB - OBJECTIVE: In a prospective study it was our intention to evaluate the reliability and the predictive value of expiratory ethanol for the early detection of the occurrence of TURP syndrome and emphasize the role of the serum levels of glycine in clinical manifestation. METHODS: We studied 30 patients scheduled for elective traditional transuretral resection of the prostate performed with subarachnoid anesthesia. Serum sodium and glycine concentrations, serum osmolality and end-expiratory ethanol levels were monitored at scheduled intervals. Continuous heart rate and blood pressure monitoring was performed during the perioperative period in the operativing room and, later, in the recovery room. Occurrence of cardiocirculatory, respiratory and neurologic symptoms were recorded. Statistics included Bonferroni's t-test and Fisher's exact test. A decision level plot for end-expiratory ethanol level was performed for the choice of predictivity criterion. RESULTS: In our population we identified three groups of patients: Group I (15 patients) in which no symptom was recorded; Group II (6 patients) in which non-specific anesthesia-related symptoms occurred; Group III (9 patients) in which TURP syndrome of various degree of severity was observed. In this group of patients changes in serum sodium and glycine concentrations, serum osmolality and end-expiratory ethanol levels were significantly different compared with the other two groups. In regard to end-expiratory ethanol levels, we identified a cut-off point at 0.05 mg/ml. In Group III two patients developed transient blindness. These patients had the highest serum glycine concentrations (> 4000 mumol/ml). Mortality was nil. CONCLUSIONS: Our data show the reliability and accuracy of end-expiratory ethanol levels as a predictive test of the occurrence of TURP syndrome. Further, we emphasize the role of serum glycine concentration in the occurrence of neurologic symptoms related to the transurethral resection of the prostate. PMID- 11494727 TI - [20 years of joint research on the carcinoma of the larynx conducted by teams from clinics in Crakow, Poznan, Warsaw and Lublin]. AB - A twenty-year scientific cooperation has been discussed (1979-1998) between four clinics for laryngology at Medical Universities in Cracow, Poznan, Warsaw and Lublin. The results of research concerning the epidemiology, diagnostics and treatment of the carcinoma of the larynx are presented. PMID- 11494728 TI - [Investigative findings on the reconstruction with tissue materials after partial laryngectomy]. AB - Between 1987 and 1999--1289 patients were treated by surgery in the Department of Otolaryngology and Laryngologic Oncology PAM in Szczecin due to the cancer of the larynx. In this group total, eventual subtotal excision of the larynx was carried out in 801 cases. In 154 cases the phonation way was restored. In this study the remaining group of 502 patients was analysed, which underwent partial aiming at restoration of all basic functions of the larynx. In the mentioned group partial vertical laryngectomy was done in 176 cases, and partial horizontal laryngectomy in 326 cases. Larynx reconstruction with the use of tissue materials was carried out in 59 cases after vertical and in 88 cases after horizontal laryngectomy. Shortcomings and advantages of the method of various tissue application were discussed (cartilage of epiglottis, vasculary based pedicle flap of thyroid gland, submaxillary salivary gland, strap muscles among others were used). PMID- 11494729 TI - [Endoscopic intranasal microsurgery in the management of cerebrospinal fluid rhinorrhea]. AB - The surgical management of cerebrospinal fluid rhinorrhoea has changed significantly after the introduction of functional endoscopic sinus surgery in the management of chronic rhinosinusitis. It gave the excellent visualization and atraumatic surgical technique in the management of eight cases of cerebrospinal fluid leaks treated in Department of Otolaryngology Medical University of Warsaw. Five of them were grafted with a pedicled middle turbinate mucosal flap. In the next three cases cerebrospinal fluid leaks came from sphenoid sinus and were stopped by free autograft of temporal fascia or abdominal fat. The indications and the advantages of the endoscopic management of cerebrospinal fluid leaks were discussed. PMID- 11494730 TI - [Primary malignant neoplasms of nasal septum in the material of ENT Department at Medical University in Gdansk]. AB - The authors presented 25 years experience of ENT Department Medical University of Gdansk in treatment 28 primary malignant neoplasms of nasal septum. The aim of the study was to analyse epidemiological (age, sex, addictions etc.), morphological and clinical data of these tumors, methods of treatment and their results. Histologically the most often recognized neoplasm was squamous cell carcinoma (15 of 28 cases--53.6%) and malignant melanoma (6 of 28 cases--21.4%). Diagnostic and clinical difficulties in this group of patients were presented. The treatment results were compared to the results obtained in the whole group of patients with malignant neoplasms of the nose and paranasal sinuses. PMID- 11494731 TI - [The assessment of compensatory mechanisms improving pharyngeal phase of deglutition after partial supra-cricoid laryngectomy. Computer tomography analysis of x-ray images]. AB - Different compensatory mechanisms after supracricoid laryngectomies of CHP and CHEP type have been observed. Wide surgical resection of anatomical structures lead to situation where laryngeal protection of lower respiratory pathway is removed. After such wide resections the lower respiratory pathway is protected against food by so called "upper closure" i.e. high placement of remaining part of the larynx and surgical movement of basis of the tongue or the tongue with epiglottis above the plane of entrance into lower respiratory pathway. Uppermost placement of the remaining laryngo-tracheal complex has been achieved by using a stabilising suture attaching trachea to sternal aponeurosis. Here, the mechanisms that protect the lower respiratory pathway and make easier pharyngeal phase of the deglutition are described. PMID- 11494732 TI - [The analysis of the relationship between integrated grade of the gray and other karyometric parameters in the precancerous lesions and laryngeal cancer]. AB - The authors assessed morphometrically the parameters describing nuclei of the normal epithelium (superficial and basal layers), precancerous lesions (dysplasia and keratosis) and laryngeal cancer in the postoperative samples of 12 patients treated in the II Laryngology Clinic Silesian Medical Academy in Zabrze. Then the relationship between the integrated grade of grey (the index of changes in DNA structure) and the others kariometric parameters was evaluated. The value of the integrated grade of grey was the smallest in the superficial layers nuclei and the biggest in the laryngeal cancer one. The evaluation of the relationship was made by using the Spearman Rang Test. The integrated grade of grey was correlated with others quantitative nuclear measurements, though the correlation power was differ in the individual morphometrical stages. PMID- 11494733 TI - [Psychological aspects of treatment of patients with head and neck cancer]. AB - One of the basic components that describe treatment success in contemporary oncology is patients' quality of life (QL) depending on specificity of the disease, treatment method, side effect of treatment, the medical prognosis and patient expectations. In reality, the QL is determined by the difference between patients' expectations and the actual health condition. The bigger the difference, the worse the evaluation of QL. Solving the crises, stress situation and starting life in a new way are strictly connected with the reshaping of personality and formation of adequate skills. It means it is necessary to create a new "picture of oneself", "self image" appropriate to the actual health condition and find the optimum way of activating personality protecting system (defence mechanisms). This task is not easy to realise; it is often necessary to introduce the psychotherapy to make it work. The main aim of defence mechanisms is the reduction of fear and other negative emotions, typical for oncological patients. However, the immature and poor defence system of "ego" can, at the same time, protect the personality unity and result in harmful behaviours. But when the "new self image" is made real, the newly learnt personality defence mechanisms are more mature, the patient becomes able to fight with the disease, is able to face the reality, which constitutes the main factor in the increase of QL assessment. To estimate the influence of psychotherapy on QL of head and neck cancer patients we have planned the introduction of psychological intervention (meetings of therapeutic groups). PMID- 11494734 TI - [The concentration of IgE antibodies in patients with cystic fibrosis]. AB - The aim of the study was to evaluate the total IgE levels and correlation between total IgE concentration in CF and allergy rhinitis patients and in healthy donors, as well as estimation of the activity of some groups of allergens by measurement of specific IgE concentration. The purpose of the study was to estimate correlation between concentration of specific IgE and skin prick tests. The concentration of total and specific IgE was measured by immunoassay. The study groups contained 16 with CF and 10 with allergy rhinitis patients, the control group contain 15 healthy donors. The highest concentration of total IgE we noted in rhinitis subjects, statistically higher than in CF patients and in the control group. The analysis of the concentration of specific IgE showed that allergens as dermatophagoides, food and grasses are the most effective in CF patients. We did not found correlation between specific IgE level and skin prick tests. PMID- 11494735 TI - [Evaluation of hearing in children with solid tumors treated by chemotherapy]. AB - Ototoxicity of cisplatin is documented in during the last years. In material of 13 children with solid tumors (neuroblastoma, nephroblastoma, rhabdomyoscarcoma) treated with cisplatin, ototoxicity was observed in 2 cases. There were presented audiological findings. The role of otoacoustic emission measurements in monitoring of cisplatin--induced ototoxicity was underlined. PMID- 11494736 TI - [Hyperlipidemia in patients with inner ear disturbances]. AB - The aim of this study was to evaluate the occurrence of hyperlipidemia in patients suffering from vertigo, and/or tinnitus and/or hearing loss of unknown origin. 48 patients (25 women and 23 men) were included into this study. All patients had a negative previous medical history of any metabolic, cardiovascular or neurological disorders. Our results were compared to the control group of 31 healthy persons (16 women and 15 men). All subjects had a complete neurootologic examination, appropriate audiometric and vestibular studies. In biochemical evaluation lipid phenotype studies were performed. Hyperlipidemia were classified according to Friedricson criteria. There were some differences in lipid phenotype and severity of hyperlipidemia between this two group. PMID- 11494737 TI - [Anatomy of the temporal bone assessed by high resolution computed tomography, endoscopy and microscopy]. AB - Visualization of all anatomical structures inside the middle ear is indispensable to perform surgical treatment. The aim of this study was the attempt to show landmarks of the middle ear employing different imaging techniques. The value of high resolution CT with 1 milimeter layers, Hopkins endoscopy and operating microscope with 16 time magnification were assessed. The primary evaluation by the CT scans were made and directed the further efforts. Endoscopy and microscopy were performed as a next diagnostic steps and allowed for tool-eye visualization of all important anatomical structures in the middle ear. The anatomy and pathology of tympanic cavity assessed by three different technics used together occurred to be much more detailed. The results were complementary and useful. PMID- 11494738 TI - [Tumors of external ear canal. Case reports]. AB - The authors described two rare cases of tumors of the external auditory meatus. All patients were male 40 and 51 years old. The tumors were removed via external auditory canal approach and extraauricle approach with successful. PMID- 11494739 TI - [A rare case of malignant lymphoma of the nasal cavity and paranasal sinuses]. AB - The head and neck region is one of the most common sites of extranodal non Hodgkin's lymphoma, but it is an infrequent tumor at the nasal cavity and paranasal sinuses site, representing no more than 5.8-8 percent of the entire malignant tumors in this localization and 0.3-2 percent of all extranodal lymphomas. A case of non-Hodgkin's lymphoma in fifty seven years-old man is reported. The patient has been admitted for curing because of returning during few last years nose bleeding, cured symptomatically. From last few weeks the wall protrusion of the right side of nose. Clinical and radiological examinations have found large pathological changes of nasal cavity and paranasal sinuses. Histological examination has shown the presence of lymphoma malignum. They also haven't found other neoplasm focuses. The sick has been curing by Co-60 radiation with a perfect effects. Now, after a years observation they haven't found a sick returning. PMID- 11494740 TI - [A rare case of adenoid cystic carcinoma of the larynx]. AB - One case of rare laryngeal tumor--adenoid cystic carcinoma in 58 year old woman was reported in this paper. Authors wanted to drew attention to few complaints and only slight dysphagia in comparison to local extensiveness of the neoplasm. Diagnostic difficulties were underline. Epidemiology, clinical course and therapy were demonstrated in laryngeal adenoid cystic carcinoma. PMID- 11494741 TI - [Middle ear adenoma: a case report]. AB - Middle ear adenoma (MEA) is a rare neoplasm with benign clinical behavior. The immunohistochemical and ultrastructural studies revealed the mixed, bidirectional mucinous and neuroendocrine character. These tumors have been known by many different names, reflecting the controversies relating to their histogenesis and differentiation. The term middle ear adenoma was proposed by Hyams and Michaels in 1976 and since this time over 100 cases were described in the literature. The most often symptoms of the MEA are the unilateral hearing loss and the tinnitus. MEA should be treated by surgery and very attentive postoperative follow-up. The prognosis of these tumors are good. The histological structure of middle ear adenoma, as well as symptomatology, treatment and clinical behavior are discussed on the basis of the literature. A case of histologically confirmed middle ear adenoma in a 34 year old women is presented. PMID- 11494742 TI - [Unusual history of the chemodectoma of parapharyngeal space]. AB - A case of chemodectoma in 58 year old woman is presented. The tumour was located in the parapharyngeal space. During preoperative arteriography and embolization a hemiparesis was noticed, probably caused by the thrombus in the cerebral medial artery. Early radiological examination of brain arteries allowed to perform specific treatment in only two hours time from the onset of the paresis. The treatment was successful--neurological symptoms have receded after 24 hours. Few days later patient was operated on--the tumour was removed with part of the vagus nerve. No serious postoperative complications were observed. PMID- 11494743 TI - [A case of malignant melanoma of the parotid gland]. AB - The case of 56 years old patient with melanoma malignant parotid gland is presented. Preoperative diagnostic examinations CCT, ultrasound, fine needle aspiration have not confirmed the neoplasm diagnosis. Fast local growth and unclear prognosis is characteristic for this tumor. PMID- 11494744 TI - [The use of benzydamine Hcl -- Tantum Verde -- in otolaryngology]. AB - Benzydamine HCL--Tantum Verde is a non-steroid drug of anti-inflammatory, pain killing and antibacterial activity. The preparation is very easy to use in the form of mouth and throat wash, nebuliser or 3 mg lozenges. It is non-toxic and does not have side-effects. It allows the patients to improve their ability to eat foods and reduces throat discomfort and pain. On the basis of literature and own observation it is to be stated that Tantum Verde has substantial usefulness in the treatment of various, inflammatory conditions, radiotherapy induced mucositis in anti-neoplastic induced stomatitis, in angina, in neoplasms with necrosis, after surgical operation of the mouth and pharynx, after intubation and endoscopic surgery in the larynx. PMID- 11494745 TI - [New criteria for detecting occult hemorrhage in screening for colorectal cancer]. AB - The author describes new varieties of fecal occult blood screening strategies in respect to old and up-to-date diagnostic methods. The limited sensitivity and specificity of the classical methods used for the demonstration of occult fecal blood necessitate the introduction of new techniques in the screening practice. The new discoveries in molecular biology are likely to improve the effectiveness of the early detection of colorectal cancers and their precursor lesions. Since the implementation of these novel techniques in mass screenings has not taken place yet, higher effectiveness can only be achieved by more specific and sensitive blood tests. PMID- 11494746 TI - [Thiazolidinediones--a new class of oral antidiabetic drugs]. AB - The discovery of a new class of oral antidiabetic drugs was stimulated by difficulties with the treatment currently available for patients with type 2 diabetes mellitus. Thiazolidinediones can lower blood glucose values due to their special insulin-sensitiser effect. In this way, these drugs seem to be very effective in the treatment of type 2 diabetic patients with characteristics of metabolic syndrome. The intracellular action caused by thiazolidinediones differs markedly from that of other oral antidiabetic drugs available. Apart from antihyperglycaemic effect, thiazolidinediones have further beneficial effects in experimental diabetes which require corroboration by clinical studies. Troglitazone was the first drug which reached the market. Unfortunately, this drug was withdrawn soon due to its hepatotoxicity. Rosiglitazone proved to be much safer in clinical studies. Pioglitazone is being tested nowadays in clinical studies. Thiazolidinediones have been already listed among oral antidiabetic drugs in international therapeutical guidelines. Nevertheless, further clinical studies and experiences are needed to determine the final exact indication of thiazolidinediones for the treatment of type 2 diabetic patients. PMID- 11494747 TI - [Serum bone marker measurements in bone metabolism disorders associated with inflammatory bowel diseases]. AB - Patients with inflammatory bowel disease (IBD) have decreased bone mineral density (BMD), which is usually much more remarkable in patients with Crohn's disease (CD) than those with ulcerative colitis (UC). The aim of the present study was to investigate the usefulness of serum beta-Crosslaps (bCL) and osteocalcin (OC) determinations to assess bone metabolism in patients with IBD. Forty-nine patients with IBD (23 UC, 26 CD) and 46 healthy controls were studied. Serum bCL and OC were measured by Elecsys immunoassay. Compared to controls (0.275 +/- 0.14 ng/ml) the mean bCL concentration was significantly higher in the CD (mean = 0.489 +/- 0.25 ng/ml; p < 0.001) and UC groups (mean = 0.439 +/- 0.3 ng/ml; p < 0.01). The mean OC concentration was significantly higher in the CD group (28.52 +/- 14.75 ng/ml) than in controls (21.42 +/- 7.43 ng/ml) but OC level was not significantly increased in the UC group (24.89 +/- 15.08 ng/ml). There was no significant difference in bCL or OC concentrations between the CD and UC groups. These results indicate that the accelerated bone resorption is not associated with increased bone formation in patients with IBD. These two marker of the bone metabolism could be a good laboratory parameter of bone pathology in patients with IBD, especially in CD. PMID- 11494748 TI - [Uniparental disomy 7 in the pathogenesis of Silver-Russell syndrome]. AB - The authors report the frequency and the clinical signs of uniparental disomy of chromosome 7 in Silver-Russell syndrome patients. A cohort of 73 families were typed with Short Tandem Repeat markers from chromosomes 7. In 6 patients maternal uniparental disomy 7 (UPD7) was detected. Summarising their data and those from the literature, an overall frequency of maternal uniparental disomy 7 of approximately 10% can be estimated. Allelic distribution in two of their maternal uniparental disomy 7 families indicates complete isodisomy whereas allelic patterns in the other four families are consistent with partial and complete heterodisomy, respectively. The clinical features of maternal uniparental disomy 7 patients do not show any deviation from the non-uniparental disomy 7 patients. Additionally, there was not hint for possible influences of iso- or heterodisomy, possibly associated with different stages of mosaicism. Their results demonstrate the necessity to screen SRS patients for UPD7 although the effect of UPD7 cannot be correlated to the SRS phenotype yet. Furthermore, an association between UPD for chromosomes other than 7 and SRS seems to be negligible. Vice versa, maternal UPD7 is not detectable in non-SRS patients. Therefore, testing for maternal UPD7 can be restricted to SRS families, searching for other UPDs in this population does not seem to be reasonable. Additionally, cytogenetic analysis should also be performed in SRS patients: identification of commonly involved chromosomal regions should allow narrowing down a SRS-relevant region. PMID- 11494749 TI - [Current treatment of anal fissure]. AB - More and more authors believe, that the cause of the chronic anal fissure is the increased anal resting pressure. It maintains a vicious circle causing ischaemia and ulcer on the anoderm. There are many traditional surgical methods treating fissures apart from the conservative ways. However, these are likely to cause significant morbidity. Chemical sphincterotomy seems to be a novel way and its most effective drugs are the nitrates. 75-80% of the patients can be cured with the topical use of these. This treatment has to be accompanied with the normalisation of the stools. If the symptoms persist after 6-8 weeks the procedure of choice is the lateral sphincterotomy. Authors evaluate the different treatments based on a literature overview and own experiences. They also suggest a treatment protocol. PMID- 11494750 TI - [Epidermoid cyst of the cecum]. AB - This paper describes a case of an epidermoid cyst arising in the anterior wall of the coecum. Epidermoid and dermoid cysts of the coecum are exceptionally rare. These cysts arise by the sequestration of squamous epithelium during embryological development, or following trauma or surgery. The true cysts are the result of inclusion or sequestration of ectodermal elements at the time of closure of the neural groove. Acquired epidermoid cysts are of traumatic or iatrogenic origin, due to implantation of the epidermis. In this reported case the patient had no previous abdominal surgery which is why this cyst represents the congenital variant of the disease. PMID- 11494751 TI - [Milestone in the treatment of diabetic dyslipidemia: the DAIS Study]. PMID- 11494752 TI - [Clostridium difficile diarrhea: frequency of detection in a medical center in Buenos Aires, Argentina]. AB - Clostridium difficile has been recognized as the most important enteric pathogen of nosocomial antibiotic-associated diarrhea (CDAD) in adults from industrialized countries. The importance of C. difficile as a cause of diarrhea in ambulatory patients appears underestimated or under-recognized. Since the 1980's, outbreaks of CDAD have been increasingly reported, but there are few data available in Argentina. We developed a retrospective study to provide some information about CDAD in our country. From July 1998 to November 1999, a total of 245 fecal specimens from hospitalized and some ambulatory patients were tested in order to confirm the diagnosis of CDAD. C. difficile cytotoxin (toxin B) was identified by detecting its cytopathic effect on monolayers of McCoy culture cells. For culture and isolation of C. difficile, stool samples were prepared by ethanol shock prior to plating onto a selective medium which contained blood, cefoxitin and fructose. Of the 245 samples, 14 (5.8%) were identified as positive by the cell cytotoxicity assay. Using the criteria of isolation of cytotoxigenic C. difficile positivity increased to 6.5% (16 samples). Thirteen of the positive results were from hospitalized patients (81.3%) and 3 (18.7%) from outpatients. The mean age of inpatients was 72.9 years (ranging from 47 to 88). All patients had received 2 or more antimicrobial agents (most of them beta-lactams) 2 months before the appearance of diarrhea. There was one patient who had received only chemotherapy. The prevalence of CDAD in this study was less than in others previously reported. This difference may be due to the fact that not all general practitioners include testing for C. difficile when the patient with diarrhea had previously received antibiotics. More educational programs should be directed to all physicians, concerning the role of C. difficile as an important enteric pathogen in patients who have undergone treatment with antimicrobial or chemotherapeutic agents. PMID- 11494753 TI - [Leptospira interrogans serovar Pomona: detection of antigenic differences among 3 regional isolates from cattle and a reference strain]. AB - Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain. PMID- 11494754 TI - [Determination of the antibacterial and antiviral activity of the essential oil from Minthostachys verticillata (Griseb.) Epling]. AB - The in vitro antiviral activity of the essential oil from Minthostachys verticillata was investigated against herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PrV). The viral inhibition was assayed employing viral plaque reduction assay. The antiviral activity of the essential oil specifically affects PrV and HSV-1 multiplication, since it was found that non toxic effects on cells were observed at the concentrations assayed. The therapeutic index values were 10.0 and 9.5 for HSV-1 and PrV, respectively. The antibacterial activity was studied using a diffusion assay and the broth tube dilution method. Gram-positive bacteria were more sensitive to inhibition by plant essential oil than the gram negative bacteria. The essential oil of M. verticillata was analyzed by gas chromatography (GC) technique. Of the six components identified in the volatile oil, pulegone (44.56%) and menthone (39.51%) were the major constituents. The antimicrobial activity can be explained to some extent by the presence of pulegone. Results suggest that further investigations concerning the isolation of the substance responsible for the antimicrobial activity and an effort to define the mechanisms of action are warranted. PMID- 11494755 TI - [AIDS-associated meningeal cryptococcosis in the Hospital Diego Paroissien from 1996-1999]. AB - A total of 148 patients with a diagnosis of HIV infection were studied in order to evaluate the incidence of meningeal cryptococcosis, including epidemic, immunologic and diagnostic characteristics. The diagnosis of cryptococcosis was carried out by direct examination with India ink and culture in Sabouraud agar of CSF in 28 patients (93.3%) and by blood cultures (lysis-centrifugation) in 2 patients (6.6%). All the isolated strains were identified as Cryptococcus neoformans. The incidence was 20.3% (30 patients). The preponderant risk behavior was endovenous drug addiction, and it was observed in 18 patients (60%). The symptomatology that prevailed was headache (87%). The median age was 28 years. At diagnosis, the immunologic impairment was severe (CD4+ lymphocyte count < 200) in 90% of patients. We found that 86.7% of patients had not completed their primary studies and only 13.3% had completed secondary studies. Although the acute mortality was high (36.7%), it was observed that all the patients who survived (24%) had been treated with anti-retroviral drugs. PMID- 11494756 TI - [Blue tongue. Update on the agent and the disease]. AB - Bluetongue (BT) is a viral disease of domestic and wild ruminants. It is particularly damaging in sheep, where up to half of infected animals may die, showing inflammation and hemorrhages of the mucous membranes of the mouth, nose, and intestines. In cattle and goats, BT rarely causes disease, however it can affect the animal's reproductive ability, so that losses are not easily estimated. Bluetongue virus spreads from animal to animal by biting insects of the genus Culicoides; and this is the reason why the disease is more prevalent in geographic areas where climate conditions are favourable for their development. The disease was first recognized in South Africa in the late 1700's, but it was not until the early 1900's that it was described in detail, and at present, epizootiology and pathogenesis studies are still being carried on. PMID- 11494758 TI - [Escherichia coli: diversity of biochemical phenotypes in aquatic environments (Santa, Fe, Argentina)]. AB - During certain environmental conditions, the floating aquatic vegetation, mainly represented by Eichhornia crassipes (water hyacinth) invade and even cover water courses assigned to recreational activities or to the supply of drinkable water. The rhizosphere of these plants constitutes an unknown biotope of bacteria of sanitary interest, possibly different from waters without vegetation and of the sediment of the same aquatic system. To verify such possibility, 206 isolated strains in MacConkey Agar (Difco) were typified and identified (78 from water, 65 from sediment and 63 from rhizosphere) using the API 20 E system (v. 4.0) and Apilab plus software (v 3.3.3), both of bioMerieux (Marcy-l'Etoile, France, 1998). Nineteen different biochemical phenotypes from E. coli were found. The 79% of the population belonged to only 7 phenotypes; the 21% remaining, to the other 12 phenotypes. Twelve phenotypes did not share the biotopes, while only 4 were in the three. These results (and those obtained by other authors who used the API 20 E system in other biotopes) suggest that it would be possible to characterize the rhizosphere using those phenotypes that are found in smaller proportion. The greatest index of diversity (H) and evenness (E) were found in the rhizosphere (H = 2.903; E = 0.874). The dendrogram (average distances and UPGMA method) reaffirms the dissimilarity in biochemical phenotypes of E. coli populations of the rhizosphere with regard to the other biotopes. The most abundant bacterial species in the three biotopes were E. coli, Klebsiella terrigena and K. pneumoniae, corresponding to 75.2% of the community. The rhizosphere differed from Serratia odorifera and from Klebsiella spp. because of its higher rate of isolation. PMID- 11494757 TI - [Degradation of plant waste by Coprinus truncorum using 2 culture methods]. AB - Degradation of yard wastes by Coprinus truncorum growing in a vertical aereated bioreactor or in flasks was studied. There was a constant decay of reducing sugars in the medium that avoided their accumulation and their possible repression of degradative enzymes. Endoxylanase activity at first showed a similar pattern in both culture conditions, with maximal activity on the 12th day, but flasks maintained a high activity thereafter. Flasks also showed a higher endoglucanase activity with a peak on the 18th day, whereas the maximal value in the bioreactor was reached on the 26th day. No Mn-peroxidase and only low values of laccase activity were found. The measurements of pH and soluble proteins during the incubation period were suitable indicators of the degradation process by C. truncorum. PMID- 11494759 TI - [Evaluation of the bacterial flora in natural corn silage]. AB - Silage is the best method for forage conservation, keeping it in a fresh condition and with high nutrient content. In order to study the natural evolution of maize silage without additives during 50 days after sealed, pH, temperature changes, number and type of the indigenous bacteria were studied every 5 days. The initial pH decreased from 6.40 to 4.10 and temperature stabilized at 26 degrees C. The mesophilic aerobic bacteria were quickly reduced across time. Enterococci were descended gradually. Clostridia remained in a low quantity. The microorganisms most frequently isolated were Escherichia coli, Clostridium butyricum, Enterococcus faecalis and Lactobacillus plantarum; this acid lactic bacteria was the principal cause of the abundant production of anaerobic fermentation of sugars in the forage reaching the pH which maintained the silage in adequate conditions. PMID- 11494760 TI - [Molecular characterization of aphthous fever virus isolated during the years 1993-1994 in Argentina]. AB - Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics. PMID- 11494761 TI - [Stability of Lactococcus lactis phages treated with sodium hypochlorite and during storage]. AB - Survival of lytic bacteriophages active against Lactococcus lactis ssp. lactis and ssp. cremoris was determined after treatment with sodium hypochlorite and during storage at 4 degrees C. Three phages were isolated from dairy plants in Argentina (ARG) and the other phages were isolated in the United States of America (US). All of them represent phages that infected cheese manufacture industries and belong to different morphological or serological groups. These phages showed higher survival in M17 broth, buffered with sodium glycerophosphate, than in trypteine soy broth (TSB). Phage populations did not decrease significantly during 14 weeks in M17 broth, whereas in TSB the titers of phage suspensions began to decline around 9 days. In addition, the effect of sodium hypochlorite was more marked in broth than in milk. A higher surviving fraction was obtained in milk, even when tenfold higher concentrations of chlorine were used. The effect of hypochlorite on phages of the same serological group was quite similar and independent of phage morphology. However, phage 137 1, which belongs to other serological group, showed lower resistance to sodium hypochlorite. Comparing the hypochlorite inactivation for ARG and US phages, it was observed that they have their own inactivation values, independently of their origin and morphological group. Long periods of time and high concentrations of chlorine were necessary to reduce the surviving fraction in milk. This indicates that hypochlorite concentrations and times of contact can be critical for the efficiency of the operative sanitization processes. PMID- 11494763 TI - Designing a better biomedical engineer. PMID- 11494762 TI - Corynebacterium diphtheriae threats in cancer patients. AB - The aim of this study was to determine the bacteriological properties of Corynebacterium diphtheriae strains isolated from bronchiole washing and cancer lesions. Bacteriological characterization included fluorescence/double sugar urease (King/DSU) screening tests, pyrazinamidase (PYZ), CAMP-reactions and radial immunodiffusion toxigenicity assay. Microorganisms produced fluorescence under ultraviolet light and were catalase positive; urea and aesculin hydrolysis negative; fermentation of glucose, maltose and sucrose and no fermentation of mannitol and xylose; PYZ and CAMP reaction negative. The API-Coryne system was used for bacterial preliminary identification at local hospital laboratory and produced numerical profiles 1010325 and 0010325 for sucrose positive C. diphtheriae var. mitis (nitrate positive) and C. diphtheriae var. belfanti (nitrate negative), respectively. The hemagglutination, adherence to glass and polystyrene assays evaluated adhesive characteristics. Strains were toxigenic and able to adhere to glass, polystyrene and human erythrocyte surfaces (titer 4). C. diphtheriae strains isolated from cancer patients expressed adhesive characteristics similar to strains isolated from immunocompetent hosts. Circulation of toxigenic C. diphtheriae continues to present a threat for children and adults including patients with cancer in hospital environment. Laboratories should remain alert to the possibility of isolation of diphtheria bacilli from adults with neoplastic disease. PMID- 11494764 TI - Genomes, man, and machines. PMID- 11494765 TI - Computing life: the challenge ahead. PMID- 11494766 TI - A model organism for genomic and postgenomic studies. PMID- 11494767 TI - Integrative data mining: the new direction in bioinformatics. PMID- 11494768 TI - Automated processing of raw DNA sequence data. PMID- 11494769 TI - Visualizing large-scale genomic sequences. PMID- 11494770 TI - Discovering the dynamics of microbial genomes. PMID- 11494771 TI - A compression algorithm for DNA sequences. PMID- 11494772 TI - Structural mining of molecular biology data. PMID- 11494773 TI - A database system for comparative genomic hybridization analysis. PMID- 11494774 TI - Two-dimensional fast magnetic resonance brain segmentation. PMID- 11494775 TI - Pharmacy education bill introduced to Congress. PMID- 11494776 TI - Report urges better end-of-life, palliative cancer care. PMID- 11494777 TI - Physicians reluctant to provide specific life-span estimates to cancer patients, researchers say. PMID- 11494778 TI - FTC cracks down on false dietary supplement ads. PMID- 11494779 TI - Almotriptan joins migraine market. PMID- 11494780 TI - Patient safety standards focus on medical-error reduction, patient notification. PMID- 11494781 TI - New England states band together in Medicaid drug program. PMID- 11494782 TI - Tablet splitting, education on NABP delegates' minds. PMID- 11494783 TI - Pharmacy and 20 years of AIDS. PMID- 11494784 TI - Potential risks and prevention, Part 3: Drug-induced threats to life. AB - Potential risk factors for and the preventability of drug-induced threats to life were studied. Case reports of adverse drug events (ADEs) published in Clin-Alert during 1977-97 were the source of information on drug-induced life threats. Patient, drug, and event variables were identified, and the causality, predictability, and preventability of each case were assessed. Data were entered into a relational database for analysis. The data indicated 846 drug-induced life threats. Seventy-four percent of the cases were assessed as definite or probable. Patients received usual or below-usual dosages in 89% of the cases. Patients tended to be middle-aged and only moderately ill. The drug categories most frequently associated with life threats were antimicrobials and central-nervous system agents. Plasma drug level monitoring should have been performed in 127 cases but occurred only in 31 cases (24%). Event types were distributed as adverse drug reactions (50%), allergic reactions (35%), drug interactions (11%), and medication errors (4%). A commercial reference classified almost half of the drug interactions associated with a life threat as posing minimal or no potential risk to the patient. Half of the life-threatening events were judged to have been preventable; about half of these could have been prevented by a pharmacist. Litigation was reported for only 1% of the cases of drug-induced threats to life; judgments and settlements averaged $1.2 million. A review of published case reports of ADEs for 1977-97 yielded information on possible risk factors for drug induced life threats and on which events may have been preventable. PMID- 11494785 TI - Potential risks and prevention, Part 4: Reports of significant adverse drug events. AB - A summary analysis of three descriptive studies of significant adverse drug events (ADEs) was conducted. Case reports of ADEs published in Clin-Alert during 1976-97 were the source of information on ADEs, including drug-induced deaths, disabilities, and threats to life. The results of the three studies were compared, and recommendations were made. During the 21-year period, 1520 significant ADEs were reported (29% resulting in death, 15% in permanent disability, and 56% in life threats). Event types were distributed as adverse drug reactions (52%), allergic drug reactions (25%), medication errors (15%), and drug interactions (8%). Only 12% of the drug interactions were classified as having highest significance by one drug information reference, while 32% of the drug interactions were unclassified. Typically, patients were 40-69 years old and relatively healthy or only moderately ill and had received usual dosages. However, 29% of the patients with a drug-induced permanent disability were less than 10 years old. Only 17% of the drugs that could have been monitored by blood level tests were so monitored. The drug categories most commonly involved in ADEs were central-nervous-system agents, antimicrobials, antineoplastics, and cardiovascular agents. The nervous, hematopoietic, cardiovascular, and respiratory systems were affected the most. Faulty prescribing was the most common reason for medication error, and wrong dosage was the most common type of error. A lawsuit was reported in 13% of the cases. Overall, 52% of the cases were judged to have been preventable; of these, 50% could have been prevented by a pharmacist. Litigation was reported for 13% of the cases; settlements and judgments averaged $3.1 million. A summary analysis of more than 1500 published case reports of ADEs for 1976-97 yielded information on possible risk factors for drug-related deaths, disabilities, and life threats and on which events may have been preventable. PMID- 11494786 TI - Multidisciplinary education and management program for children with asthma. AB - A multidisciplinary program for managing asthma in a pediatric population is discussed. A coordinated, multidisciplinary program for managing asthma in children was initiated in November 1997 at a U.S. Army medical center. The program, designed to improve care and decrease hospitalizations for asthma, was pharmacist managed and pulmonologist directed and was implemented by pediatricians. Patient education was provided by a pediatric clinical pharmacist or a nurse case manager; providers also received intensive education. Follow-up occurred at predetermined intervals and included asthma education, discussion of expectations and goals, analysis of metered-dose-inhaler and spacer technique, and assessment of compliance. Between November 1997 and January 1999, 210 inpatients were screened for asthma. One hundred seven were believed to have asthma and received inpatient asthma counseling and teaching. Of these 107 patients, 79 were enrolled in the program and monitored in the ambulatory care setting. Seventy-one (90%) of the 79 program enrollees were not rehospitalized during the ensuing two years. The number of children admitted to the hospital for asthma decreased from 147 in 1997 (a rate of 3.2 per 1000 population) to 93 in 1998 (2.1 per 1000) and to 87 in 1999 (1.9 per 1000). A multidisciplinary approach to the management of children with asthma may reduce hospitalizations of such patients. PMID- 11494787 TI - Stability of Adderall in extemporaneously compounded oral liquids. AB - The short-term stability of Adderall in three extemporaneously compounded oral liquids was studied. Three suspensions of Adderall 1 mg/mL were prepared from commercially available 10-mg Adderall tablets with Ora-Sweet, Ora-Plus, and a 1:1 mixture of Ora-Sweet and Ora-Plus. Each suspension was stored in the dark in a stability chamber at 25 degrees C and 60% relative humidity for 30 days. The stability of the active drug (a mixture of levoamphetamine and dextroamphetamine salts) in each of the three vehicles was determined immediately after preparation and at 10, 20, and 30 days by using gas chromatography-mass spectrometry (GCMS). No significant changes in concentrations of either amphetamine isomer occurred during the 30-day study period. Visual inspection of samples revealed no changes in color or odor. Extemporaneously compounded liquid oral formulations of Adderall 1 mg/mL in Ora-Sweet, Ora-Plus, or a 1:1 mixture of Ora-Sweet and Ora Plus were stable for at least 30 days at 25 degrees C and 60% relative humidity. PMID- 11494788 TI - Visual compatibility of sodium nitroprusside with other injectable medications given to pediatric patients. PMID- 11494789 TI - Interventions and documentation for drug-related problems in Dutch community pharmacies. PMID- 11494790 TI - Comparison of eptifibatide and abciximab with decision analysis. PMID- 11494791 TI - ASHP guidelines on managing drug product shortages. PMID- 11494793 TI - Doc Holliday, D.D.S.: 150th anniversary. PMID- 11494792 TI - The 2000 C.T. Rowland Award orthodontic case report. PMID- 11494794 TI - The endodontist's painful dilemma. Response to ethical dilemma #36. PMID- 11494797 TI - New technology helps detect and treat tooth decay. PMID- 11494795 TI - Oral and maxillofacial pathology case of the month. Renal-osteodystrophy induced macrognathia. PMID- 11494798 TI - Potassium nitrate-fluoride reduces bleaching sensitivity, report says. PMID- 11494799 TI - Scheme funding for paedodontic dentures. PMID- 11494800 TI - Bonding agents: adhesive layer thickness and retention to cavity surfaces with time. AB - This study assessed bonding agent thickness, cover and the influence of long-term storage on bonding agent retention to enamel and dentine surfaces in cut occlusal cavities in 46 human molar teeth. Two specimens were etched and set aside. The remaining specimens were divided into two equal groups and treated with either Optibond or Scotchbond Multipurpose Plus (SMPP) up to the adhesive stage. Thereafter two specimens from each group were stored for 0, 1, 3, 7 and 14 days and 1, 2, 3, 6 and 12 months in 1% NaCl. After storage the cavity surfaces were examined in a scanning electron microscope (SEM) and scored as to bonding agent cover. One specimen from each treatment was then embedded in resin, two sections prepared midway through each cavity, polished and re-examined in the SEM to measure bonding agent thickness at 13 sites along the cavity surface. Both bonding agents showed highly variable and significant (P < 0.05) bonding agent cover and layer thickness according to cavity site, SMPP more so than Optibond. Pooling of SMPP adhesive was apparent in cavity angles. Mean film thickness was significantly different between Optibond (221 +/- 130 microns) and SMPP (118 +/- 106 microns). There was no significant difference in bonding agent thickness between long- and short-term storage. PMID- 11494801 TI - Fluoride concentration of bottled drinking waters. AB - The use of bottled water and beverages may be a significant source of systemic fluoride and can therefore be considered as a risk factor for dental fluorosis in young children. The aim of this study was to determine the fluoride content of commercially available bottled drinking waters and to report on the accuracy of the labelling of fluoride concentration. Thirty brands of bottled water, classified as either spring (N = 19) or mineral (N = 11) water were evaluated. A fluoride ions-elective and a fluoride reference electrode were used to measure the fluoride concentrations. The average reading for each brand was compared with the fluoride content printed on the label. Only 56.7% (N = 17) of brands tested mention the fluoride concentration on the label, but 73.3% (N = 22) had a tested fluoride concentration of less than 0.3 ppm. Of the 8 brands testing higher than 0.3 ppm fluoride, 1 did not have the fluoride concentration labelled, while for another the tested fluoride concentration was much higher than the concentration printed on the label. When prescribing fluoride supplements, dentists should be aware of the fluoride content of bottled waters used by child patients, especially brands with a concentration higher than 0.3 ppm. PMID- 11494802 TI - Occlusal reconstruction of a collapsed bite by orthodontic treatment, pre prosthetic surgery and implant supported prostheses. A case report. AB - The loss of mandibular molars can result in a 'collapsed bite' owing to tilting of teeth adjacent to the gap and overeruption of maxillary molar segments. The lost interarch and interdental space must be regained before prosthetic reconstruction. This case report documents the treatment of a patient by orthodontic, surgical and prosthetic means. The teeth were orthodontically aligned to meet predetermined surgical and prosthetic requirements. The surgical phase comprised a posterior segmental maxillary osteotomy and one-stage placement of three large-diameter implants in the mandible. Finally, the occlusion was restored with mandibular implant-supported prostheses. PMID- 11494803 TI - The effect of a single educational input given to school teachers on patient's correct handling after dental trauma. AB - The aim of this study was to evaluate the influence of mailed guidelines and seminars to school teachers on self-care actions taken by children after trauma. Data was collected 6 months before and 5 months after provision of the guidelines. Particulars of school children who sought dental consultation, types of injuries, the medium used to store avulsed teeth and the time elapsed from injury to dental consultation were recorded. A non-significant increase was observed in a number of children who sustained tooth avulsion from primary schools that belonged to a seminar group and from nursery schools that belonged to a mailed guidelines group. There were no significant differences before and after provision of guidelines in the number of patients with soft-tissue injury, periodontal injury and injury to hard dental structures. Of the patients who sustained tooth avulsion, 36 (54%) brought the involved tooth to the dental clinic, (29%) sought dental consultation later than 7 hours after injury. None of those patients stored the avulsed tooth in a recommended transport medium. The findings of this study indicate that a single educational input to school teachers is not enough to promote children's self-care after injury. PMID- 11494804 TI - Short communication: Prevalence of suspected latex allergies among a group of South African dentists. PMID- 11494805 TI - [Lasers in dentistry. Part A--Development, characteristics, structure, principles of operation and types of lasers]. AB - Laser technology is developing very quickly. New Lasers with a wide range of characteristics are available today and are being used in the various fields of dentistry. The search for new devices and technologies for dental procedures was always challenging and in the last two decades much experience and knowledge has been gained. The purpose of this series of articles published in the journal of the Israel Dental Association in three parts is to present the Israeli dentist a comprehensive and contemporary overview which will assist him in understanding the potential of the use of laser technology for clinical applications in dentistry. The first part will describe the history and development of lasers in dentistry, the characteristics and structure of lasers as well as their principles of operation. A description of the different types of lasers used today in dentistry will be presented in the first part. The second part will describe the interaction of lasers with biological tissues and their effect on the soft and hard tissues of the oral cavity as well as on the dental pulp. Laser safety in the dental practice will also be discussed. The third part will focus on clinical applications of Lasers in Dentistry: pits and fissures sealing, dentin desensitization, cavity preparations, composite curing, surgical procedures and tooth whitening. Current status of using lasers in periodontics, endodontics and dental implantology will be discussed and possible future clinical applications of lasers in dentistry will be presented. PMID- 11494806 TI - [Single-stage implants in areas of maxillary sinuses that were augmented with allograft. Case reports]. AB - Demineralized freezed dried bone allograft (DFDBA) is known as a bone inductive material, and used widely in periodontal and bone regeneration procedures. DFDBA can also be used for sinus floor augmentation prior or with implant placement. The present manuscript described cases of single-stage implants that were used successfully in areas where the maxillary sinus was augmented with DFDBA simultaneously with implant placement (Case 1) or in a second procedure, 18 months following grafting of the sinus (Case 2). These cases demonstrated the ability to use single-stage implants in the augmented maxillary sinus, in a separated or simultaneous procedure. PMID- 11494807 TI - [Inferior alveolar nerve repositioning in implant surgery]. AB - Severe resorption of the posterior mandible possesses one of the most difficult restorative challenges to the implant surgery today. This resorption may prevent the placement of dental implants without the potentially damage to the inferior alveolar nerve. To create the opportunity of insertion dental implants of adequately length in those cases, the technique of nerve repositioning has been advocated. The purpose of this article is to describe two cases of nerve repositioning combined with placement of dental implants. Both cases showed appropriate postoperative healing without damage to the inferior alveolar nerve. The inferior alveolar nerve repositioning technique seems to be an acceptable alternative to augmentation procedure prior to dental implants placement in cases exhibiting atrophic posterior mandibular ridges. PMID- 11494808 TI - [The orthodontic treatment of children with disabilities]. AB - Due to the medical progress in the past decades, the life expectancy and quality of the physically and/or mentally disabled children have been much improved. While this has facilitated their entry into society, it has also created greater demands on our profession, from those who care for them. A displeasing dental appearance may have a significant emotional impact on an individual's well being and can bias judgments of social acceptability, ability and personality. Gross misplacement of the teeth occurs more often and with much greater severity in the disabled child than in the general population. The increased concern for facial appearance and oral functions has generated an increased demand for orthodontic treatment, but the difficulties imposed by the handicapping conditions have made many practitioners believe that these patients are not amenable to treatment. At the Center for the Treatment of Craniofacial Anomalies in the Orthodontic Department of the Hadassah School of Dental Medicine, Jerusalem we have been treating children with disabilities for several years. An investigation of the referral patterns of our patients has revealed ignorance of dentists, physicians and social workers of what can and may be done to improve the dental and facial appearance of these children. This finding was the principle reason for this article, in which we will show how successful delivery of treatment is possible for many of the disabled. PMID- 11494809 TI - [Effect of professional training on dental health attitudes of Israeli dental students]. AB - Few studies have been published regarding the importance of oral hygiene education for dental students and little is known about the influence of dental education in dental schools on students' attitudes to the subject. The objective of the present research was to examine the changes that occur in the attitudes of Israeli dental students toward their dental health during the course of their professional training. The research was based on a questionnaire developed at The Hiroshima University, Japan (The Hiroshima University Dental Behavioral Inventory, HU-DBI), and provides a quantitative estimate of the students' attitudes to their dental health. Similar studies have been carried out in several countries (Japan, Australia, Indonesia and Finland). This study was performed on students from the two dental schools in Israel, The Maurice and Gabriela Goldschleger School of Dental Medicine, Tel Aviv University and The Faculty of Dental Medicine at Hebrew University, Jerusalem. Students from all six academic years (1st-6th) were requested to complete the questionnaire within two months, from the beginning of the academic year that started in October 1999. While no significant differences could be detected in the students' attitudes with regard to the dental school of their origin, female students (from both schools) showed a significantly better attitude than their male colleagues throughout the years. Results showed a significant improvement in the students' attitudes through the years of their professional training, especially between the 1st year and the clinical years (5th and 6th) of their studies. This can be related to the teaching curricula in the dental schools in Israel that emphasizes clinical issues in the two last years of study. When comparing the results of this study to similar ones that were conducted in other parts of the world, certain differences become apparent. In spite of the gradual improvement in the Israeli students' attitudes during the course of their studies, their initial attitudes (1st year) as well as their final attitudes (6th year), were the lowest compared to their colleagues from other countries (Japanese, Australian and Finnish students). Apparently, the improvement in the Israeli students' attitudes is not satisfying and a higher level of improvement should be expected. The attitudes of the Israeli students should be further improved by means of comprehensive programs that emphasize the importance of personal oral hygiene and dental health. Hopefully this will enable future improvement in the dental health of their patients, as well as improvement in the community dental health in Israel, in general. PMID- 11494810 TI - [The Israel Periodontal Society position statement: The use of lasers in periodontal treatment]. PMID- 11494812 TI - Dentistry and evidence-based care. PMID- 11494811 TI - Periodontal medicine in the next millennium. AB - Dentistry and medicine have, to a great extent, been somewhat separated during the last 160 years, despite the fact that they have the same patients in common. We have now reached a period in our history when research is bringing medicine and dentistry closer together with data that are cogent to physicians and dentists and, most importantly, to the patient. A new paradigm is emerging with regard to susceptibility to periodontal disease, its etiology, and pathogenesis. Definite relationships between the oral disease and systemic conditions show that some risk factors are a concern for periodontal disease and certain systemic diseased such as cardiovascular disease. Periodontal diseases and systemic diseases--this is a two-way street. It is becoming clear that the dentist needs to know more about systemic diseases, and the physician needs to increase his or her knowledge of oral diseases. We may see an increase in oral microbiology testing for patients with periodontal disease. We also will see more dentists doing glucose testing as well as other tests in their practices. Physicians and dentists working more closely together, more patients with systemic diseases will be managed more successfully, and patients will benefit from predictable treatment regimens to save their dentition. PMID- 11494813 TI - Information on the Internet found to be usually accurate but also incomplete. AB - As hospitals, health systems, and health plans look toward strengthening their Web sites, they may want to keep in mind findings from a recent study that notes that while much of the information that consumers find on the Internet is accurate, many users find it incomplete and difficult to understand. PMID- 11494814 TI - Organizations show greater interest in meeting quality standards. PMID- 11494815 TI - Creating portals to quality: how the Internet is changing health care delivery to consumers. AB - The number of Americans visiting health care sites on the Internet has steadily increased every year. However, while many say the sites are helpful, more are looking to these sites to change from static to interactive--to provide information tailored to their needs, to enhance self-care, to encourage better provider communication, and to aid in lifestyle management. Some health care organizations are beginning to provide new services that promise to give e-health a new look. PMID- 11494816 TI - Tilt of Senate to Democrats speeded debate on patients' rights. PMID- 11494817 TI - Compensating the pharmacy services manager. PMID- 11494818 TI - Too many patients, too few physicians. PMID- 11494819 TI - HMOs need to share gains of DM programs. Physician are more likely to buy in if they see better outcomes--and financial rewards that go with them. AB - Perhaps nowhere in medicine does the friction between physician and HMO translate into lost revenue more noticeably than in disease mnagement, where lack of doctor buy-in is a perennial complaint. The author, a former medical director knowledgeable about all aspects of DM, offers two methods by which health plans can overcome this obstacle. PMID- 11494820 TI - AMA slams managed care even as leadership splinters. PMID- 11494821 TI - Moving beyond a punitive mind-set. Interview by Patrick Mullen. PMID- 11494822 TI - Satisfaction and patient outcomes of a telephone-based nurse triage service. AB - PURPOSE: Describe patient satisfaction and patient-reported outcomes after voluntary use of a telephone-based nurse triage service. METHODS: A random sample of symptomatic callers who contacted the triage service in 1999 was identified. A computer-assisted telephone survey was conducted, resulting in a response rate of 58.9 percent and a sample size of 35,374. SUMMARY: Overall satisfaction with the service was 90.4 percent and did not vary greatly when stratified by demographic and health status characteristics. Of all callers who reported following the triage recommendation to use self-care instructions while monitoring the condition for change (n = 12,037), 11.5 percent scheduled an office visit and 1.5 percent used hospital emergency-room (ER) services for further care. CONCLUSIONS: Overall satisfaction with telephone-based nurse triage services was high and did not vary substantially by caller characteristics. PMID- 11494823 TI - Companies search for ways to pass costs on to employees. PMID- 11494824 TI - Stark allows federal regulators to review claims records fully. PMID- 11494825 TI - Slow, steady drop seen in employee health benefits. PMID- 11494826 TI - A new, encapsulated glass ionomer for band cementation. PMID- 11494827 TI - Micro-implant anchorage for treatment of skeletal Class I bialveolar protrusion. PMID- 11494828 TI - What is a practice worth? PMID- 11494829 TI - TOMAC: an orthognathic treatment planning system. Part 2 VTO construction in the horizontal dimension. PMID- 11494830 TI - Avoiding demineralization and bite alteration from full-coverage plastic appliances. PMID- 11494831 TI - Use of lingual brackets for deep-bite correction. PMID- 11494832 TI - Upper extremity overuse injuries in swimming. A discussion of swimmer's shoulder. AB - Treatment of shoulder pain includes the following: 1. Avoid all painful activities. 2. A 2-week course of nonsteroidal anti-inflammatory medication and ice. 3. Decreased anterior capsule stretching and increased posterior capsule stretching. 4. Increased rotator cuff exercise with emphasis on external rotators. 5. Scapular-positioning muscle exercises and increasing body roll. Shoulder pain can be prevented by the following: 1. Avoid all painful activities, and notify coach of shoulder pain immediately. 2. Do not use nonsteroidal anti inflammatory medications or ice on a chronic basis. 3. Spend equal time stretching the posterior and anterior capsules. 4. Perform general rotator cuff exercises. 5. Perform scapular-positioning muscle exercises, with emphasis on body roll. [figure: see text] Shoulder pain in swimmers is common and can be debilitating. Most of the pain is caused by instability, which stems from swimming-specific demands that increase performance but decrease shoulder stability. These sport-specific demands are (1) increased shoulder range of motion, (2) increased internal rotation and adduction strength, and (3) prolonged, fatiguing, shoulder-intensive training. Instability leads to [figure: see text] inflammation and pain and can become a self-perpetuating process. Treatment consists of patient education, cessation of all activities that cause pain, activity modifications to increase shoulder stability, and pharmacologic treatment of the inflammation. In patients who do not improve using this regimen, surgery can be of benefit, either to reduce capsular laxity or to remove chronic inflammation and scar tissue. The patient must be aware of the risk of decreased performance. PMID- 11494833 TI - Overuse injuries of the upper extremity in tennis players. AB - Several overuse injuries can affect the upper extremity of tennis players. A thorough understanding of anatomy and knowledge of these common conditions and their treatment facilitates the management of these athletes. PMID- 11494834 TI - Overuse injuries of the upper extremity in baseball. AB - The shoulder and elbow are prone to many overuse injuries in baseball. Injury to the muscles or ligaments leads to pain and loss of effectiveness in competition. Although many of these disorders respond to conservative management, surgery is sometimes necessary to provide pain relief and restore function. Unfortunately, overuse injuries that require surgery still are career-threatening injuries. Correct diagnosis and treatment increase the probability of a throwing athlete returning to competition. PMID- 11494835 TI - Overuse injuries of the upper extremity in golf. AB - Golfers are athletes, for golf is a sport. To play it well, one must have athletic ability, strength, agility, coordination, and endurance. The golf swing is physically demanding and has contributed to overuse injuries in the upper extremity. The wrist, elbow, and shoulder are the most frequently injured joints. PMID- 11494836 TI - Upper extremity injuries associated with strength training. AB - Most injuries sustained during strength training are mild strains that resolve with appropriate rest. More severe injuries include traumatic shoulder dislocations, tendon ruptures of the pectoralis major, biceps, and triceps; stress fractures of the distal clavicle, humerus, radius, and ulna; traumatic fractures of the distal radius and ulna in adolescent weightlifters; and compressive and stretch neuropathies. These more severe injuries are usually the result of improperly performing a strength training exercise. Educating athletes regarding proper strength-training techniques serves to reverse established injury patterns and to prevent these injuries in the first place. Recognizing the association of anabolic steroid use to several of the injury patterns further reinforces the need for medical specialists to counsel athletes against their use. With the increasing use of supplements such as creatine, the incidence and nature of strength-training injuries may change further. Greater emphasis on the competitive performance of younger athletes undoubtedly will generate enthusiasm for strength training at earlier ages in both sexes. The importance of proper supervision of these young athletes by knowledgeable persons will increase. As the popularity of strength training grows, there will be ample opportunity to continue to catalog the injury patterns associated with this activity. PMID- 11494837 TI - Impingement syndrome. AB - Pain from rotator cuff pathology is a common problem in upper extremity athletes. Impingement of the rotator cuff tendons on the subacromial arch is an important cause. Primary age-related degeneration and instability, however, should not be forgotten as causes for cuff pathology. Conservative treatment is successful in many athletes. Surgical subacromial decompression is reserved for resistant cases. Although often helpful, surgical treatment generally requires a long recovery for throwing athletes and can be associated with residual symptoms. PMID- 11494838 TI - Biceps tendinitis and subluxation. AB - Since the 17th century, the long head of the biceps tendon as a source of shoulder pain and its functional significance has been a source of debate. Although the term tendinitis is commonly used, overuse tendon injuries infrequently demonstrate inflammatory cells; instead, degenerative changes resulting from the failure of self-repair usually are found. Bicipital tendinitis or bicipital tenosynovitis is most often secondary to impingement beneath the coracoacromical arch. Primary bicipital tendinitis and tendinitis secondary to instability are possible, however. Through a careful history, physical examination, and appropriate imaging studies, the clinician can establish the diagnosis of disorders of the biceps tendon Arthroscopic evaluation greatly improves the diagnosis and treatment of biceps tendon and related shoulder pathology. Although the exact functional role of the biceps tendon remains incompletely defined, a growing body of evidence supports its role as a stabilizer of the glenohumeral joint. This stabilizing function should be incorporated into the treatment of biceps tendon disorders. Routine tenodesis has been replaced by a more individualized approach, taking into consideration physiologic age, activity level, expectations, and exact shoulder pathology present. New repair techniques are under development, and preservation of the biceps-labral complex is now preferred when possible. PMID- 11494839 TI - Pronator syndrome. AB - Although pronator syndrome is often misdiagnosed and does not occur as frequently as carpal tunnel syndrome, its clinical features are well described, and the diagnosis should be made if the clinical features are understood and reasonable suspicion exists. The differentiating features between carpal tunnel syndrome and pronator syndrome should be understood, and evaluation for one site of compressive neuropathy of the median nerve always should include the other potential sites. When the correct diagnosis is made, pronator syndrome can be successfully treated nonoperatively or surgically, if necessary. PMID- 11494840 TI - Medial epicondylitis and cubital tunnel syndrome in the throwing athlete. AB - Medial epicondylitis and ulnar nerve problems are common in the throwing athlete, resulting from the tremendous valgus stress that occurs during the acceleration phase of pitching. They are too often ignored as a cause of medial pain in the throwing athlete in favor of the diagnosis of ulnar collateral ligament injury. The latter seems to be in vogue, and there is a virtual epidemic of reconstructive surgery to address this diagnosis. The question is whether more medial pain is caused by the former two diagnoses than the latter. We need to have a better understanding of the natural history and differential diagnosis of medial side pain, which, I hope, will come with time. Nonetheless, medial stress injuries occur in the throwing athlete, and can cause inflammation of the adjacent anterior capsule flexor pronator mass, the ulnar collateral ligament, and the ulnar nerve. This review highlights these problems, their anatomy, diagnosis, and management. PMID- 11494841 TI - Lateral epicondylitis. AB - Satisfactory treatment of lateral epicondylitis results from correct diagnosis followed by a well-controlled operative or nonoperative treatment program. Many options for nonoperative and operative treatment exist for lateral epicondylitis. More study is needed on outcomes of both nonoperative treatment and operative treatment so that each patient can attain maximal improvement. Balanced assessments of specific patient populations, along with definitions of the optimal treatment for each group, are required. This will allow physicians to integrate the available information and improve patient care. PMID- 11494842 TI - Osteochondritis dissecans of the humeral capitellum. Diagnosis and treatment. AB - Elbow pain seen in the at-risk athlete, such as a baseball player (in particular, a pitcher) or gymnast, should raise suspicion for OCD. OCD of the humeral capitellum remains a difficult problem to treat. Once radiographic changes are obvious, long-term studies suggest that half of affected individuals will be symptomatic. Currently, the key to successful treatment is early detection. Gymnasts, in general, fare worse in returning to sport. The reason is not entirely clear but is likely related to the amount of force directed across the elbow and the nature of the sport. Pitchers sometimes can be returned, but to another position. Radiographs remain the screening test of choice but can give the surgeon a false sense of security, because changes early in the disease process may not be obvious radiographically. Views at 45 degrees flexion and contralateral elbow views are helpful. The advent of MR imaging now allows the practicing orthopaedic surgeon to assess very early lesions effectively that might otherwise be underappreciated on radiographs. With more advanced lesions, radiographic findings are more obvious and demonstrate the more classic capitellar fragment with a surrounding zone of lucency. MR imaging is this setting is helpful in assessing the overlying articular cartilage and, hence, the stability of the fragment. In the absence of obvious loose bodies or mechanical symptoms, rest is the first step in treatment. If symptoms persist, then operative intervention is indicated. About half of these patients heal with nonoperative treatment. Pretreatment assessment of fragment viability has not traditionally been incorporated into the treatment algorithm. Recent anecdotal evidence suggests that stability and viability of a fragment can be assessed using intravenous contrast. Knowledge of the fragment viability could allow those lesions likely to heal without surgical intervention to be distinguished from those requiring surgical intervention. The procedure should be chosen based on the size of the lesion and the integrity of the subchondral bone. Subchondral drilling and microfracture can only resurface defects and cannot reconstitute subchondral bone. Autologous chondrocyte implantation has limited ability to address subchondral bone loss, whereas autograft and allograft osteochondral transplantation can restore subchondral bone. Most authors would agree that there is no role currently for reduction and fixation of long-standing, free loose bodies. No consensus exists regarding acute dislodging of an in situ loose fragment. Long-term results after radiographic changes are present suggest a degenerative course in about half the patients. Whether the newer techniques of cartilage resurfacing will significantly impact the natural history of this process remains to be seen. PMID- 11494843 TI - Wrist and hand overuse syndromes. AB - Overuse syndromes in the wrist or hand can occur from repetitive use of the wrist and hand or from recurrent direct trauma to the hand area. Tendinitis syndromes due to overstretching or shear stress are seen commonly on both the extensor and flexor sides of the wrist. Overuse syndromes also can take the form of neurovascular syndromes, resulting in compression syndromes of the median, ulnar, and superficial branch of the radial nerve in the wrist area and trauma to the ulnar and digital vessels supplying the hand. Treatment in most cases involves rest with splinting, icing, and NSAIDs in acute cases, although surgical decompression is indicated in chronic or recurrent cases. PMID- 11494844 TI - Stress fractures of the upper extremity. AB - Stress fractures are common injuries in the athletic population. Although much of the published literature has focused on lower extremity stress injuries, these injuries also occur in the upper extremities. Stress injuries of bone result from repetitive loads smaller than would be required to cause an acute fracture. As bone is repetitively stressed, it behaves like any solid substance. If deformity occurs within its elastic range, it returns to its original configuration. If stressed into its plastic range, permanent deformity occurs, and microfractures propagate, causing structural failure and complete fracture of the involved bone. High clinical suspicion is required for diagnosis because of historical and physical features can be vague. Plain radiographs are often inconclusive, but bone scans and MR imaging usually help elucidate the diagnosis. Most upper extremity stress injuries will heal with nonoperative management. In rare situations these injuries can progress to nonunion, which requires surgical correction. PMID- 11494845 TI - Rehabilitation concepts and supportive devices for overuse injuries of the upper extremities. AB - Rehabilitation of overuse injuries requires the health care professional to understand normal and common pathologic mechanics of the patient's sport or physical activity. Evaluation of the total mechanics of the body, not just the local area presenting with discomfort, is important. [figure: see text] The cause of the presenting injury can be a dysfunction at a distant site. Rest and supportive devices are helpful in allowing the athlete to return to a normal activity level. A progressive reconditioning program needs to address strength and flexibility issues and endurance and eccentric exercises. PMID- 11494847 TI - Volume changes accompanying interaction of ligands with nucleic acids. PMID- 11494846 TI - Calorimetric techniques in the study of high-order DNA-drug interactions. PMID- 11494848 TI - Calculating sequence-dependent melting stability of duplex DNA oligomers and multiplex sequence analysis by graphs. AB - The analytical methods for characterizing DNA sequence-dependent thermodynamic stability have been reviewed. A set of n-n sequence stability parameters is presented. Examples in which these values are used to calculate the thermodynamic stability of short duplex DNA oligomers are presented. The problem of determining sets of isothermal sequences is addressed by representing DNA sequences as graphs. Representing DNA sequences by a graph descriptor with special mathematical properties minimizes the computational difficulty of determining the number of DNA sequences with identical predicted thermodynamic stability. This is achieved by replacement of a whole set of sequences by a single representative. Applications of this concept were demonstrated for sequences assembled from individual bases and sequences assembled from oligomeric blocks. PMID- 11494849 TI - Thermal denaturation as tool to study DNA-ligand interactions. PMID- 11494850 TI - Kinetics of binding of Hoechst dyes to DNA studied by stopped-flow fluorescence techniques. PMID- 11494851 TI - Surface plasmon resonance biosensor analysis of RNA-small molecule interactions. PMID- 11494852 TI - Scanning force microscopy of nucleic acid complexes. AB - SFM is a viable effective method for determining the mode of binding, the extent of binding, and the site of binding of intercalators to nucleic acids. Establishing the presence of a groove-bound ligand can be achieved either by competitive binding experiments with a well-defined intercalator (minor groove) or by changes in apparent contrast (major groove). In our opinion, SFM has an important role in resolving the structural polymorphisms for small molecule-DNA complexes. Application of these assays in the study of polyintercalator molecules is currently underway in our laboratory. SFM is an important, new tool in the study of protein-DNA complexes. New insights into the structure and function of these complexes are enabled by real-time visualization. Currently the temporal resolution of the SFM limits the degree to which definitive rate data can be determined. Several binding and unbinding events could take place in the time it takes to acquire one image. New developments in SFM technology will allow faster scanning and will improve the temporal resolution of so-called SFM movies. To this end, the Hansma group is developing small cantilevers and improved optical deflection systems to enable intermittent imaging at scanning rates of 1.7 sec per image. These improvements will enable SFM visualization of complex biological processes as they occur, one molecule at a time. PMID- 11494853 TI - Nuclear magnetic resonance studies of drug-DNA complexes in solution. PMID- 11494854 TI - X-ray crystallography of DNA-drug complexes. AB - Here we have stressed important differences between protein and DNA crystallography. Crystal growth and data collection methodologies are not directly transferable between the two subfields. In addition, we note that analysis of symmetry and packing of DNA crystals can be useful and a uniquely aesthetic exercise. PMID- 11494855 TI - Molecular modeling of drug-DNA complexes: an update. PMID- 11494856 TI - Analysis and interpretation of ligand-DNA binding isotherms. AB - Binding studies provide information of fundamental and central importance for the complete understanding of ligand-DNA interactions. Studies of ligand binding to long natural DNA samples, to synthetic deoxypolynucleotides of simple repeating sequence, and to oligonucleotides of defined sequence are all needed to begin to understand the interaction in detail. Binding studies provide entry into the thermodynamics of the DNA interactions, which in turn provides great insight into the molecular forces that drive the binding process. This chapter summarizes both model-dependent and -independent approaches for the analysis and interpretation of binding isotherms, and should serve as a concise guide for handling experimental data. PMID- 11494857 TI - Peptide nucleic acid targeting of double-stranded DNA. PMID- 11494858 TI - Drug interaction with triple-helical nucleic acids. PMID- 11494859 TI - Measurement of covalent drug-DNA interactions at the nucleotide level in cells at pharmacologically relevant doses. PMID- 11494860 TI - Targeting DNA through-covalent interactions of reversible binding drugs. PMID- 11494861 TI - Chemical cross-linking of drugs to DNA. PMID- 11494862 TI - Drug-RNA footprinting. AB - RNase I and RNase T1 can be used to obtain high-quality footprinting information for paromomycin binding to a 176-mer RNA from the packaging region of HIV-1 (LAI). Controls and scanning procedures are necessary for quantitation of autoradiographic data, so that footprinting plots showing cutting behavior as a function of drug concentration can be used to identify binding sites and regions of altered structure on the 176-mer. From the RNase I footprinting results the primary paromomycin binding sites on the 176-mer are on the main stem and on the stem of SL1, but noncontiguous sequences may be involved in the same binding event. Strong enhancements in cleavage with added drug are also observed, indicating drug-induced structural changes. Drug binding may cause linker regions between stem-loops of the 176-mer to change structure, possibly providing a site or sites for additional drug binding. Because drug binding changes the structure of the packaging region, which may alter its function, paromomycin analogs with enhanced specificity for HIV psi RNA have potential as a new class of agent for treating AIDS. PMID- 11494863 TI - Footprinting methods for analysis of pyrrole-imidazole polyamide/DNA complexes. PMID- 11494864 TI - High-resolution transcription assay for probing drug-DNA interactions at individual drug sites. PMID- 11494865 TI - Use of DNA molecules substituted with unnatural nucleotides to probe specific drug-DNA interactions. PMID- 11494866 TI - Drug interactions with nucleosomes and chromatin. PMID- 11494867 TI - Simultaneous measurement of binding constants and unwinding angles by gel electrophoresis. PMID- 11494868 TI - Locating cobalt-binding sites on DNA using restriction endonucleases. PMID- 11494869 TI - Exploiting anthracycline scaffold for designing DNA-targeting agents. PMID- 11494870 TI - Design, synthesis, and characterization of polyintercalating ligands. PMID- 11494871 TI - Targeting telomeres and telomerase. PMID- 11494872 TI - Rapid, high-throughput engineering of sequence-specific zinc finger DNA-binding proteins. PMID- 11494873 TI - DNA relaxation and cleavage assays to study topoisomerase I inhibitors. PMID- 11494874 TI - In vitro human immunodeficiency virus type 1 integrase assays. PMID- 11494875 TI - Use of Xenopus egg extracts to study effects of DNA-binding drugs on chromatin assembly, nuclear assembly, and DNA replication. PMID- 11494876 TI - Linear and circular dichroism of drug-nucleic acid complexes. PMID- 11494877 TI - Rapid screening of structurally selective ligand binding to nucleic acids. PMID- 11494878 TI - [Electrophysiologic parameters determining the success of radiofrequency ablation of atrioventricular reentry tachycardia]. AB - The main objective was to assess electrophysiological parameters which may be important for effective radiofrequency ablation of accessory atrioventricular pathways. In a group of 66 patients a total of 490 applications of radiofrequency current were made, incl. 68 which led to successful severing of the accessory pathways. The ablations were successful in 65 patients (98.5%). The potential of the accessory pathways was present in effective ablations in 19 cases of 68 applications (28%), while in ineffective ablations it was present in 13 cases of 421 applications (3%, p < 0.001). Stability of the electrogram was found in all 68 effective ablations (100%), while in ineffective ablations a stable electrogram was found in 248 cases of 323 applications (77%, p < 0.001). In unstable electrograms the ablation was not effective in any of the patients. The V-delta interval was significantly longer in successful ablations (16 +/- 14.2 ms) than in unsuccessful ones (3 +/- 19.4 ms, p < 0.001). The ventriculoatrial interval was in successful ablations significantly shorter (57.5 +/- 22.5 ms) than in unsuccessful ones (74.5 +/- 20.9 ms, p < 0.001). In 90% applications the ventriculoatrial interval was less than 70 ms. Of 10 evaluated parameters the potential of the accessory pathway, stability of the electrogram, V-delta interval (equal or < or = 0) and short ventriculoatrial intervals (equal to or < or = 70 ms) proved important parameters for successful ablation of accessory pathways. PMID- 11494879 TI - [Heart failure in hypertensive patients with left ventricular hypertrophy]. AB - Left ventricular hypertrophy (LVH) is supposed to be a useful marker of cardiovascular complications during the course of hypertension. Authors compared the presence of heart failure, left ventricular diastolic dysfunction and chronic atrial fibrillation in hypertensive patients with and without left ventricular hypertrophy defined by echocardiography. Hospital records of 192 hypertensives treated in our medical department during years 1996-1999 were analysed. Left ventricular hypertrophy was defined by echocardiography (Penn convention) as left ventricular mass index > 134 g/m2 in men and > 110 g/m2 in women. Presence of LVH was found in 128 patients (mean age 65.9 years), absence of LVH in 64 patients (mean age 64.8 years). Both groups of hypertensives were matched by demographic parameters, by the presence of hyperlipidemia, by smoking habits. Hypertensive patients with left ventricular hypertrophy were more often treated by ACE inhibitors. There were statistically significant more patients with heart failure, left ventricular diastolic dysfunction and chronic atrial fibrillation in LVH-positive patients than in LVH-negative once. There was also statistically significant lower ejection fraction (50.3 +/- 11.4% vs 56.5 +/- 7.4%) in LVH positive patients than in LVH-negative once. Left ventricular hypertrophy in patients with hypertension brings usually a complicated course of the disease with a high contribution to the development of chronic heart failure. PMID- 11494880 TI - [Acute hemorrhage of the upper digestive tract--personal experience]. AB - The authors submit a retrospective investigation of 50 patients hospitalized at the intensive care unit of the Medical gastroenterological department, Faculty Hospital Brno treated in 1999 with the diagnosis of acute haemorrhage into the upper digestive tract. In the investigated group the most frequent cause of haemorrhage was portal hypertension (21 patients, 32.8%) and a peptic gastroduodenal lesion (15 patients, 23.4%). During the investigation period 12 patients died (18.8%), 6 developed haemorrhage as a complication of a serious condition (decompensated cirrhosis of the liver). In haemorrhage from oesophageal and gastric varicosities pharmacotherapy is equally important as endoscopic intervention. PMID- 11494881 TI - [Implantable cardioverter-defibrillators in the prevention of sudden cardiac death]. AB - The aim of this work is to characterize and analyse the spectrum of therapies delivered from implantable cardioverter-defibrillators (ICD), to evaluate their effectivity and to mark the most effective types of antitachycardia stimulations (ATP), cardioversion (CV) and defibrillation (CD). To compare our results with references and give precautions of trouble-shootings. Our patients had implanted ICDs according to standard criteria for ICD implantations. Before discharge from hospital we performed predischarge test of their ICD. Than we followed them periodically each three months. We have observed 72 ICD pts (55 M, 17 F) in the mean age of 62.7 +/- 12.2 years the with mean LVEF was 0.37 +/- 0.11. The mean follow-up was 21 +/- 12.8 months. Each examination was managed through anamnesis of symptoms accompanying the beginning of arrhythmia, the sensation of ICD therapy by patient, followed by interrogation of the ICD memory. All obtained episodes were analysed. During the follow-up 1023 episodes of malignant ventricular arrhythmias were detected and effectively terminated. 7 pts died. During the therapy the ATP reached 83% in comparison with CV, CD which reached only 17%. The dominating symptoms were palpitations and presyncopes. In comparison with initial arrhythmias leading to implantations of ICDs (ventricular fibrillations for most of the cases--54%) the significantly higher number of spontaneous episodes were caused by monomorphic ventricular tachycardias VT (92.0%). We had no sudden cardiac death in our pts. In the indicated pts with a high risk of sudden arrhythmic death, the ICD therapy is characterized as very effective and is associated with high safety, low discomfort and when up to date algorithms for detection being used, then only adequate part of the inappropriate therapies occurs (10% patients, 3.2% from the number of episodes). The therapy by implantable cardioverter-defibrillators has had an important role in treating pts with life-threatening ventricular arrhythmias. PMID- 11494882 TI - [Intestinal permeability in relatives of patients with Crohn's disease]. AB - The etiology of Crohn's disease remains unknown, one of theories is the damage of bowel wall with the impairment of small bowel barrier. In patients with Crohn's disease was found high level of small bowel permeability in dependence of disease activity. The aim of our study was to measure small bowel permeability in first degree relatives of the patients with Crohn's disease. In 14 first-degree relatives of Crohn's disease patients (7 men, 7 women) there was measured small permeability using lactulose, mannitol and D-xylose. Sugars were investigated in 5 hour collected urine and examined using capillary gas chromatography. The absorption of all the sugars--lactulose, mannitol and D-xylose did not differ from healthy controls. The small bowel permeability index (lactulose/mannitol) did not differ from healthy controls too (0.023 vs. 0.019, p = 0.451). These preliminary results confirms the theory, that there is no damage of small bowel barrier in first degree relatives of Crohn's disease patients. PMID- 11494883 TI - [Pitfalls in the diagnosis of infectious endocarditis]. AB - A group of 63 patients with infectious endocarditis (IE) (1991-1998) was subjected to a detailed retrospective analysis. The authors investigated the age of the patients, site of IE, type of infectious agent, incidence of coinciding diseases or interventions in the close premorbid period of IE, size of vegetation, incidence of embilizations, heart failure, indications for vital early cardiosurgery, number of deaths. From the clinical analysis it may be concluded: 1. There is a new group of patients with IE, who before development of the disease have no manifest cardiac disease who however frequently suffer from another coinciding disease; 2. There is a significant increase of dextrolateral IE (frequently "pacemaker" IE); 3. The ratio of staphylococcal strains in the etiopathogenesis of IE is rising; 4. The mean age of the affected patients is rising (as well as the age scatter); 5. IE is unfortunately frequently diagnosed only after embolization of the vegetation (mostly cerebrovascular attacks) in obscure febrile conditions. PMID- 11494884 TI - [Apoptosis, its mechanisms and medical significance. I. Definition of apoptosis and its progression at the cellular level]. AB - Apoptosis is a one example of genetically-regulated cell death. It is characterized by the phylogenetically highly conserved activation of specific genes and proteins followed by energy-consuming proteolytic autodestruction. Apoptosis seems to be physiological and highly selective mechanism to eliminate either old or injured cells in organisms. It is in contrary to mitosis. A highly complex combination of various stimuli both extracellular and intracellular origins is required for induction of apoptosis together with preserved oxidative phosphorylation. PMID- 11494885 TI - [Apoptosis, its mechanisms and medical significance. II. Disorders of apoptosis regulation and their relation to the development of diseases]. AB - Study of apoptosis seems to be relevant to clinical medicine nowadays. Dysregulation of apoptosis is under the certain circumstances the cause of various pathological processes in organism. The numerous methods to analyse apoptosis are now well established. It is very likely that our better understanding of apoptosis could lead to improvement in diagnosis and treatment of diseases such as immunopathology, cancer, and others. PMID- 11494887 TI - [Endoscopic findings and clinical diagnosis in gastroenterology]. PMID- 11494886 TI - [Non-thrombotic pulmonary embolisms]. AB - In the absence of intracardiac or intrapulmonary shunts, the pulmonary microvasculature traps all embolized particles greater than 10 microns. In the vast majority of cases these emboli are venous thrombi which lodge in larger pulmonary arteries, resulting in obstructive phenomena rather than primarily being injurious to lung parenchyma. This review deals exclusively with nonthrombotic pulmonary embolism which involves substances that lodge predominantly in the pulmonary microvessels and cause a true endothelial and parenchymal injury, as well as obstructive phenomena. The article focuses on fat embolism, amniotic fluid embolism, gas embolism, septic emboli, and tumour pulmonary embolism. PMID- 11494888 TI - [Diagnosis and therapy of anemia in chronic diseases]. AB - The article deals with contemporary views on anaemia associated with chronic diseases. The authors present the definition of this nosological unit, draw attention to its high incidence in clinical practice and fact that it is frequently mistaken for iron deficiency anaemia. The authors submit a review of the most frequent diseases which cause the development of this type of anaemia and analyze the role of activation of the system of cellular immunity, the monocyte-macrophage system, agents of the cytokine network in inhibition of proliferation and differentiation of erythroid precursors, reduced production of endogenous erythropoietin with a reduced sensitivity of erythroid progenitor cells to its action and impaired iron homeostasis and inhibition of its reutilization. Special attention is devoted to diagnostic and differential diagnostic criteria in relation to other types of anaemia caused by impaired haeme synthesis and some secondary multifactorially conditioned types of anaemia. More detailed attention is paid to the diagnostic value of evaluating serum levels of soluble transferrin receptors and explanation of the asset of calculation of the transferrin receptor/ferririn index as a sensitive indicator of latent sideropenia as well as the Fe-absorption test using low oral iron doses. Part of the paper is also an account of contemporary possibilities of treatment including the use of the recombinant form of human erythropoietin, and attention is drawn to the unsuitability and pitfalls of iron therapy in this type of anaemia. PMID- 11494889 TI - [Electrocardiographic changes in patients with acute pancreatitis. Case report and review of the literature]. AB - Transient electrocardiographic changes in patients with acute pancreatitis are well known in the literature. Mostly these changes are in the form of T-wave inversion, ST-segment depression, and rarely ST-segment elevation without the presence of coronary artery disease. We report a patient, in whom electrocardiographic changes mimicked acute inferior myocardial infarction with subsequent evolution of Q-waves in the inferior leads and ischaemia in the anterior wall. To the authors' knowledge, this is the first report documenting the evolution of Q-waves on surface ECG in the absence of myocardial necrosis verified by postmortem examination in the patient, who died of cardiorespiratory failure and massive haemoperitoneum as a complication of ongoing acute necrotizing haemorrhagic pancreatitis. The authors also discuss diagnostic and therapeutic options in patients with acute pancreatitis and ECG pattern of acute myocardial infarction. Acute pancreatitis may mimic acute myocardial ischaemia (or infarction) or these two diseases may be present at the same time. In differential diagnosis, selective coronarography might be helpful and it allows also immediate revascularisation. Administration of thrombolytic therapy in such patients is not safe and might end up with fatal consequences. PMID- 11494890 TI - [Transcatheter embolization of a coronary fistula in the treatment of ischemic heart disease]. AB - The authors present the case of a 71-year-old female patient with clinical signs of ischaemic heart disease where they diagnosed a coronary fistula between the ramus interventricularis anterior and the trunk of the pulmonary artery. Because of the marked symptomatology they made a transcatheter embolisation of the fistula by introduction of embolisation spirals which led to the elimination of ischaemic symptomatology. Because of the unusual character, pretentious technical aspects and possible risks this operation is indicated only in adult symptomatic patients. PMID- 11494891 TI - [Prevention of risk of cross-infection through the respiratory system during anesthesia]. AB - The authors present a modern concept of prevention of cross contamination during forced ventilation of the lungs in the course of total anesthesia and operation. The optimal complex of measures of individual protection of a patient includes hygienic measures, use of sterile respiratory systems (better disposable), obligatory use of respiratory filters between the intubation tube and T-piece of the narcosis and respiration device; hydrophobic folded membrane filters should be preferred. Studies carried out at Anesthesiology Department of Research Center of Surgery of Russian Academy of Medical Sciences indicate that BB 22-15 filters (Pall Corporation) reliably protect patients from cross-contamination realized through respiratory gases and biological fluids (blood, saliva, condensate) and save heat and humor. Hydrophobic folded membrane filter BB 22-15 is highly effective, which recommends it for wide use. PMID- 11494892 TI - [Principles of prevention of pneumonia associated with the use of artificial ventilation of the lungs in resuscitation and intensive care units]. AB - Pneumonia ranks among the most incident complications associated with forced ventilation of the lungs (FVL). Its incidence depends on FVL duration and according to published reports varies from 9 to 70%. Pneumonia deteriorates the prognosis and essentially increases the mortality in intensive care wards. Based on published reports and their own experience, the authors formulate the fundamentals of prevention of pneumonia in patients on FVL: use of intubation tubes with low-pressure cuffs; minimum duration or no procedures involving the intubation tube cuff blowing off; regular sanitization of the tracheobronchial tree and oropharynx; use of devices for removal of tracheobronchial secretion in the closed contour and of disposable catheters; inhalation of bronchomucolytics and antibiotics through a nebulizer; patient's position in bed with elevated head part; rigid approach to prescription of antacide drugs and H2-receptor blockers; decontamination and regulation of intestinal function; antibiotic therapy with consideration for the results of bacteriological studies; no or minimum exposure to procedures involving the respiratory contour seal opening; use of sterile gloves; use of disposable respiratory contours and hydrophobic bacterial filters instead of humidifiers. PMID- 11494893 TI - [Cuffed oropharyngeal airway: a new tool for airway management]. AB - The aim of the study was to evaluate the use of cuffed oropharyngeal airway (COPA) in spontaneously breathing patients (n = 162) under total anesthesia and to compare the new method with the laryngeal mask (LM) (n = 172). COPA provided adequate well-hermetized airway in operations under total anesthesia on spontaneously breathing patients. COPA can serve as an alternative to LM in patients with spontaneous respiration, as no appreciable differences in the incidence and type of complications in the two groups were observed. COPA extends the anesthesiologist's potentialities in ensuring reliable patency of airways, is simple, and requires no special training. PMID- 11494894 TI - [Correction of cardiorespiratory disorders in laparoscopic cholecystectomy]. AB - Carboxyperitoneum and traditional forced ventilation of the lungs have a negative impact on external respiration function during laparoscopic operations, leading to impairment of the ventilation device, pressure rise in airways, and decrease in oxygen diffusion and carbon dioxide release. This leads to accumulation of carbon dioxide in the blood and tissue with a trend to development of acidosis of mixed origin. Cardiovascular changes during laparoscopic cholecystectomy manifest by hypertension and tachicardia in the presence of increased central venous pressure and total peripheral vascular resistance, decreased stroke and cardiac indexes, decreased right-ventricular diastolic function, increased pressure in the pulmonary artery, and deceleration of venous bloodflow in the inferior and superior venae cavae. The most rational variant of forced ventilation of the lungs in laparoscopic cholecystectomy is high-frequency injection ventilation, which appreciably attenuates the negative effect of carboxyperitoneum on central hemodynamics, gas exchange, and external respiration function. The optimal variant of total anesthesia in laparoscopic cholecystectmy is endotracheal combined narcosis with diprivane and fentanide. The key factor in the choice of forced ventilation protocols is the maintenance of adequate gas exchange in the lungs in the presence of the lowest possible mean pressure in the airways. PMID- 11494895 TI - [Prolonged epidural analgesia with ultracaine for labor pain relief]. AB - Prolonged epidural analgesia (PEA) for labor pain relief and effect of this method of conduction blockade on the main vital systems of mother and newborn were studied. PEA with ultracain was used in 486 women aged 18-38 years. Puncture and catheterization of the epidural space was made at the level of the second third lumbar vertebrae; a single dose of 1% ultracain was 1.0-1.2 mg/kg. Effects of this method on the hemodynamics, external respiration function, autonomic status, and newborn status were evaluated. PEA with ultracain proved to be highly effective and safe for the mother and exerted no depressive effect on the newborn. Positive characteristics of ultracain are the minimum latent period, long (86.8 +/- 5.1 min) duration of sensory blocking, and no side effects. PMID- 11494896 TI - [Anesthesia in the early postoperative period in patients who had undergone extensive surgical interventions on the colon with plastic repair]. AB - Reconstructive plastic (n = 24) and repair operations (n = 23) on the colon were performed in 47 patients at Coloproctology Research Center (Moscow). The patients were divided into 2 groups administered different anesthesias and postoperative analgesias: total intravenous anesthesia (TIA) and combined epidural anesthesia (CEA). Adequate analgesia by CEA and early activation of patients after the operation promoted earlier (by 2 days) recovery of the motor function of the intestine than after TIA and traditional intramuscular analgesia with a narcotic analgetic. CEA ensured complete blocking of nociceptive pulsation and neurovegetative protection a with lower doses of narcotics during the postoperative period and can be used in this type of operations and postoperative management. CEA prevented gastrointestinal pareses, improved microcirculation and metabolic processes at the zone of intervention, and decreased the incidence of postoperative complications. PMID- 11494897 TI - [Lipid peroxidation and content of medium weight molecules in heart surgery with artificial blood circulation]. AB - Malonic dialdehyde (MDA), level of medium-weight molecules, and chemiluminescence (CL) of the blood were evaluated in 24 coronary patients during and after aorto mammary-coronary bypass operations. Time course of MDA concentration, content of medium-weight molecules, and CL extinction rate were similar in all the patients, but the values were higher in the patients with a complicated course of the postoperative period. Significant difference in CL intensity at the beginning of total anesthesia in patients with uneventful and complicated course of the postoperative period recommend this method for prediction of reperfusion complications. In further studies we intend to determine the diagnostically significant differences in the studied parameters and to distinguish the most informative parameters for each stage, for more effective prevention of oxidative stress in cardiosurgical patients subjected to aortocoronary bypass operations. PMID- 11494898 TI - [Intraoperative reinfusion of erythrocyte mass in neuroanesthesiology]. AB - Automated reinfusion of autoerythrocytes prepared from blood lost during removal of tumors was the main component of transfusion therapy in 49 patients (52 operations) with brain tumors. All patients developed massive blood loss of 0.5-5 TCB during the intervention, reinfusion device cell saver C.A.T.S 2-02 (Fresenius, Germany) was used. Various aspects of clinical application of this method are discussed, its efficiency and factors affecting it are analyzed. Special attention is paid to time course of hemostasis values during automated reinfusion and the problem of tumor contamination of reinfused suspension. This latter problem was solved by using the last-generation leukocyte filter RC-400 Klev (Pall, Germany). Automated reinfusion of autoerythromass effectively compensated for massive intraoperative blood loss, on condition of correction of hemostasis disorders by fresh frozen plasma and purification of reinfused suspension from tumor cells by filtering through leukocytic filters. Moreover, our results indicate that utilization of cell saver is obligatory for some patients with supermassive hemorrhages. PMID- 11494899 TI - [Esterase-metabolized remifentanil hydrochloride opioid (Ultiva-TM), a new step towards the solution of the problem of regulation of analgetic components of general anesthesia]. AB - Hemodynamics, gas exchange, velocity of psychomotor recovery, pain intensity during and after laparoscopic cholecystectomy were studied in patients anesthetized (total intravenous anesthesia) by two methods: 1) remifentanyl and propofol, forced ventilation of the lungs, and myoplegia (n = 21, ASA I-III) and 2) fentanyl and propofol, forced ventilation of the lungs, and myoplegia (n = 18, ASA I-III). Total intravenous anesthesia based on remifentanyl was characterized by stability of hemodynamic and gas exchange parameters at all stages of the intervention. This method ensured a smooth and rapid course of resuscitation, characterized by absence of signs of central respiration depression, predictable normalization of psychomotor status, and low incidence of factors provoking postoperative nausea and vomiting, on condition of their prevention. Further studies of remifentanyl are needed in interventions of different degree of traumatism and within the framework of different protocols of postoperative analgesia. PMID- 11494900 TI - [Changes in cerebral blood flow, intracranial and cerebral perfusion pressure in neurosurgical patients during anesthesia induced with diprivan]. AB - Effect of diprivan on cerebral blood flow, intracranial (ICP) and cerebral perfused pressure (CPP) was studied in neurosurgical patients during induction narcosis. The labored perfusion syndrome was observed in all patients during the initial stage. Infusion of diprivan at the initial stage of narcosis in a dose of 2 mg/kg led to a decrease in arterial pressure, mean arterial pressure, CPP, and ICP. During intubation ICP decreased by 3-7 mm Hg. At the stage of anesthesia stabilization ICP decreased by 27.3%, mean arterial pressure by 10%, CPP by 6%, linear blood flow velocity in the mid-cerebral artery of the intact hemisphere by 28.1% and in the involved hemisphere by 39.9%, with simultaneous leveling of cerebral blood flow between them. PMID- 11494902 TI - [Electrochemical detoxication of the lymph in the treatment of patients with suppurative-resorptive endotoxicosis]. AB - A new method for lymph detoxication in patients with pyoresorptive endotoxicosis is proposed. The method is based on electrochemical oxidation of the lymph, is simple and cheap. After 4-h exposure of the lymph with 0.04% sodium hypochlorite the concentration of the main toxic metabolites appreciably decreased, while the levels of total protein and leukocytes changed negligibly. Electrochemical detoxication of the lymph was used in the treatment of 13 patients with pyoresorptive endotoxicosis and led to improvement of the clinical status and rapid decrease in the levels of the major toxic metabolites, which was particularly expressed 3 days after the treatment. No negative effects were observed. Hence, electrochemical detoxication of the lymph appreciably improved the results of treatment of patients with cholestatic endotoxicosis. PMID- 11494901 TI - [High-volume hemodiafiltration in the treatment of sepsis and multiple organ failure: 2 methods of the elimination of TNF-alpha]. AB - Permanent hemodiafiltration (PHDF) as a method for treating patients with sepsis and multiple organ failure (MOF) corrects the severity of generalized inflammatory reaction, which is caused by hyperproduction of bioactive substances. Cytokines can be eliminated from circulation by 2 methods of kidney replacing therapy: convection and adsorption on hemofilter membrane. Despite the slight adsorption clearance, our results indicate that experimental data not always correspond to the clinical situation. Pronounced cytokinemia persists in patients with sepsis and MOF during PHDF. In addition to correction of the main hemostasis parameters, kidney-replacing therapy eliminates an appreciable amount of TNF-alpha and other proinflammatory cytokines. Estimation of TNF-alpha balance indicates its good adsorption on the surface of hemodiafilter membrane. PMID- 11494903 TI - [Use of plasmapheresis for correction of metabolic disorders in patients with surgical sepsis]. AB - Twenty-four patients aged 16-67 years with surgical sepsis developing as a result of pancreonecrosis, gynecological diseases, urological sepsis. closed abdominal injury, and suppurative inflammation of soft tissues were examined. Endotoxemia caused pronounced disorders in metabolism, particularly purine metabolism, and the associated lipid peroxidation processes. Plasmapheresis exerted a positive effect in patients with high concentration of medium molecular-weight polypeptides (more than 0.32 arb. units) in the blood, decreasing the mortality by 40.5%. At lower concentrations of these polypeptides in the blood (less than 0.32 arb. units) plasmapheresis is inefficient. PMID- 11494904 TI - [Use of hypertonic sodium chloride solution for correction of arterial hypotension in patients with endogenous intoxication]. AB - Reaction to infusion of 7.5% sodium chloride solution was studied in 81 patients with endogenous intoxication during development of critical arterial hypotension which could not be prevented or arrested by routine high-dose therapy. The reaction was considered adequate on condition that arterial pressure increased no higher than the safe level, subsequent maintenance of hemodynamics was effective, toxin binding with plasma albumin was increased, diuresis increased, and hypernatriemia was transitory in the presence of general improvement of patient's status. Such a reaction was observed in 83% patients. The efficiency of this method is determined by indications for low-dose therapy and probability of adequate reaction to infusion. PMID- 11494905 TI - [Possibilities of reparative regeneration of the tracheal epithelium under conditions of ozone therapy in experimental animals]. PMID- 11494906 TI - [Development of anesthesiology and resuscitation service in the Saratov region]. PMID- 11494907 TI - [Characteristics of the organization of anesthesiology-resuscitation service in large multi-profile hospitals]. PMID- 11494908 TI - [Metabolic complications of parenteral nutrition]. PMID- 11494909 TI - [Low-molecular-weight heparin - enoxaparin (clexane) in the prevention of thromboembolic complications in surgical patients]. PMID- 11494910 TI - [Anesthesiology in the surgical treatment of supraventricular tachyarrhythmias]. AB - Different anesthesias were used in 197 patients operated for supraventricular tachiarrhythmias through a transvenous access. Hypnoanalgesia based on preventive injection of a potent nonnarcotic antiinflammatory agent xefocame (lornoxicame), drip infusion of propofol (2-3 mg/kg/h), and bolus injection of dormicum under conditions of spontaneous respiration proved to be the best method. PMID- 11494911 TI - [Therapy of the adrenocortical carcinoma with Lysodren (o,p'-DDD). Therapeutic management by monitoring o,p'-DDD blood levels]. AB - BACKGROUND: o,p'-DDD (1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)-ethane), which leads to a cytotoxic necrosis of the adrenal glands, currently is therapy of choice for metastasized adrenocortical carcinomas. Clinical experience is still poor, but most studies demonstrate an increment of survival time in patients treated with o,p'-DDD after incomplete surgery. The therapeutic range is close and therefore dosage is difficult, mainly based on clinical signs. Methods for routine determination of o,p'-DDD are not yet broadly available. PATIENTS AND METHOD: We developed a method for the determination of o,p'-DDD and report our experience with the monitoring of serum levels of o,p'-DDD in our patients. Nine patients have been included, eight patients with metastasized adrenocortical carcinoma were treated with o,p'-DDD. RESULTS: At time of evaluation six of eight patients had deceased, 27.9 +/- 25.1 months after recurrence of the disease. Time of survival for all patients was 28.2 +/- 22.0 months since diagnosis of recurrence. In three patients the target dose of 9-10 g/d could not be reached due to the clinical situation. The serum levels of these patients were low (6.3 +/- 4.2 micrograms/ml). Mean survival time was significantly longer for those patients who reached serum levels above 14 micrograms/ml in comparison to those who failed to reach such high levels (41.3 +/- 16.2 vs 6.3 +/- 3.6 months, p < 0.01). The dose which was necessary to reach high levels was individually different. All patients developed adrenocortical insufficiency. Other side effects were fatigue (seven patients), gastrointestinal problems and elevated liver enzymes (six patients each), changes in blood count (five patients) and central nervous disorder (four patients). All patients developing intolerable side effects had very high serum levels of o,p'-DDD (> 20 micrograms/ml). CONCLUSIONS: Our data confirm that the efficacy of a therapy with o,p'-DDD as well as the risk to develop intolerable side effects depend on the serum levels of o,p'-DDD. Monitoring of o,p'-DDD therapy by measuring serum levels of o,p'-DDD helps to adapt this therapy individually, avoiding serious side effects as well as to realize antitherapeutical resistance. Determination of o,p'-DDD serum levels helps to decide over intensification or cessation of therapy. PMID- 11494912 TI - [Clinical value of urinary pyridinium crosslinks as osteoporosis markers: evaluation of a population survey of vertebral osteoporosis]. AB - BACKGROUND: The diagnostic value of biochemical parameters of bone turnover for the diagnosis of osteoporosis is open to discussion. We investigated whether the determination of crosslinks, bone type I collagen degradation products, correctly identifies osteoporotic subjects. PATIENTS AND METHOD: In a sample of 370 individuals recruited at random within a population survey of vertebral osteoporosis, urinary concentration of total pyridinoline and desoxypyridinoline were determined by HPLC. Standardized lateral X-rays of the thoracic and lumbar spine were taken and evaluated morphometrically using the method described by Eastell-Melton. RESULTS: Crosslink excretion was significantly higher in female but not in male individuals with vertebral deformities as defined by Eastell. The specificity of these biochemical parameters with regard to radiological osteoporotic alterations was 76-81%, but the sensitivity was 32.4-42.9% only. CONCLUSION: Pyridinoline and desoxypyridinoline reflect the process of bone degradation which leads to vertebral deformity. Crosslinks are specific markers of bone resorption and provide a valid parameter in the diagnosis of osteoporosis. The low sensitivity indicates that the measurement of pyridinoline and desoxypyridinoline is less suitable for screening purposes, but may be useful in confirming presence or extent of osteoporosis. PMID- 11494913 TI - [Fluoroquinolones: utilization in renal and urogenital tract infections]. AB - The purpose of this review is to give an introduction in the class of fluoroquinolones, which has become a large class of substances, and to discuss the use, role, and place of these drugs in the treatment of urinary tract infections. The classification of the fluoroquinolones contains four groups. Antibacterial spectrum, pharmacokinetics, and indications are the main criteria. The various fluoroquinolones differ in their antibacterial spectrum, the antibacterial activity, and pharmacokinetic properties. Therefore selection, use, and dosage regime of the substances should be carried out under respective clinical requirements and medical points of view. The physician has to consider the specific indications for each substance in each individual case. Fluoroquinolones are suitable antibacterial agents for various urinary tract infections and are a valuable and useful addition to the antimicrobial treatment. PMID- 11494914 TI - [HIV-lipodystrophy syndrome]. AB - CLINICAL FINDINGS: The HIV-associated lipodystrophy syndrome is reported with an increasing incidence predominantly in HIV-seropositive patients on antiretroviral therapy. The core of the lipodystrophy syndrome consists of fat loss in face, extremities and buttocks with or without a visceral fat accumulation, hyperlipidemia and a disturbed glucose metabolism are considered essential parts of the syndrome too. Additional metabolic abnormalities are regarded as syndrome related. PATHOGENESIS: Currently identified risk factors associated with the development of lipodystrophy are antiretroviral treatment (duration, number and kind of drugs), low CD4 cell count, higher age and metabolic abnormalities. The currently favored hypotheses to explain the lipodystrophy syndrome are essentially based on the assumption that antiretroviral treatment is the cause of the disorder. TREATMENT: The short-term benefit of antiretroviral treatment is greater than the disadvantages of the lipodystrophy syndrome, in the long term, however, the cardiovascular risk associated with the metabolic disturbances may result in an increased mortality on its own. The current approach to treatment includes modification of antiretroviral therapy and specific treatment of symptoms. PMID- 11494915 TI - [Osteoporosis and multiple pregnancy--a case report with positive outcome]. AB - BACKGROUND: Pregnancy-associated osteoporosis is a rare condition. Due to the rareness of pregnancy-associated osteoporosis, no guidelines concerning an adequate therapy exist. However, since many antiresorptive drugs are potentially teratogenous, the therapeutic approach is limited. CASE REPORT: In a 30-year-old patient, pubic fracture occurred during her first pregnancy. Osteodensitometry revealed a distinct osteoporosis. The bone density improved under therapy with sex hormones, alendronate, 1,000 mg calcium and 1,000 IU cholecalciferol daily, but still remained osteoporotic when the patient again became pregnant 3 years later. During her triplet pregnancy the patient was treated with 3,000 mg calcium and 1,500 IU cholecalciferol daily. After delivery the bone density remained at the same level as immediately before the second pregnancy. CONCLUSION: Regarding the nonoccurrence of the expected considerable bone loss with this treatment the efficacy of this therapeutic approach during pregnancy warrants further study. PMID- 11494916 TI - [48-year old woman with diffuse pulmonary calcifications after allogeneic kidney transplantation. A rare manifestation of tertiary hyperparathyroidism]. AB - HISTORY: A 48-year-old woman was hospitalized because of the presence of asymptomatic bilateral pulmonary infiltrates in the conventional chest radiograph. An antibiotic treatment administered 3 months before had been without any effect on the radiological aspect. The patient had had an end-stage renal disease of unknown cause and was on hemodialysis since 1993. 1997 she received a cadaveric renal transplant. INVESTIGATIONS: Computed tomography scan detected the infiltrates as diffuse calcifications mainly at the lung bases. Extended arterial vascular calcifications were also found. Since approximately 4 years calcium phosphorus product and parathyroid hormone had been increased. TREATMENT AND COURSE: Because of the long-term increased calcium-phosphorus product and the high parathyroid hormone levels we interpreted the pulmonary calcifications as a result of the tertiary hyperparathyroidism. After parathyroidectomy calcium phosphorus product and PTH levels normalized, but our patient suffered from increasing disturbances of the peripheral blood circulation induced by the vascular calcifications, necessitating amputation of finger and toe tips. CONCLUSION: Patients with renal failure should undergo regular controls of calcium, phosphorus and parathyroid hormone when creatinine clearance is below 50 ml/min/1.73 m2. Only an early medical treatment can prevent the various complications induced by the secondary or tertiary hyperparathyroidism. If medical therapy fails, parathyroidectomy has to be done. Especially before renal transplantation the extent of hyperparathyroidism has to be carefully analyzed and treated aggressively, if necessary by surgery before the date of transplantation. PMID- 11494917 TI - [Suspected liver metastases in a patient with porphyria cutanea tarda]. PMID- 11494918 TI - [New series on the topic of quality management]. PMID- 11494919 TI - [Quality management within the requirements of current health care policy; introduction--political and institutional aspects]. AB - The implementation of a DRG-based payment system for hospitals and the introduction of managed care features in health care plans covering care for in- and outpatients lead to an increased cost pressure on the inpatient sector. Quality assessment and quality management programs are considered a useful tool to support the process of restructuring and reorganizing hospitals as an reaction to this situation. However, changed incentives introduced by the new payment system, as risk selection or decrease of length of stay, have the potential to worsen quality in health care. As a result, the recent health care reform included a large number of quality control issues such as quality assessment, internal quality management, clinical guidelines, and evidence-based medicine. From the viewpoint of internal quality control programs, the situation is controversial, because the internal and external motivation for these programs are not identical. While the internal reason to invest in quality management is given by rationalizing processes, development of the organization and, in some cases, to optimize documentation, health legislation is primarily interested in avoiding quality problems. Health legislation is now faced with the task to build up a framework which supports quality of health care as a major argument in competition on the health care market. PMID- 11494920 TI - Time, change, and family practice. PMID- 11494921 TI - Varicella. To be [vaccinated] or not to be: that is the question! PMID- 11494922 TI - Direction of health care. PMID- 11494923 TI - Maternal obesity and risk of neural tube defects. AB - QUESTION: One of my patients is taking olanzapine for schizophrenia. She has gained a lot of weight, which, I understand, often happens with some of the new atypical antipsychotics. Due to her weight gain, she failed to notice she had become pregnant. Is she at risk? ANSWER: Experience with olanzapine is relatively slight, but available prospective data do not show increased teratogenic risk. Adiposity, on the other hand, is associated with increased risk of neural tube defects. Only some of this risk can be reduced by folate supplementation. PMID- 11494924 TI - Ophthaproblem. Chemical injury to the eye. PMID- 11494925 TI - Medicolegal file. Demands from a patient's lawyer. PMID- 11494926 TI - Just the berries. Allergy to local anesthetics. PMID- 11494927 TI - Cyclooxygenase (COX-2) selective inhibitors. Any better than NSAIDs? PMID- 11494928 TI - Physicians certified in family medicine. What are they doing 8 to 10 years later? AB - OBJECTIVE: To determine field of medicine and location of a cohort of physicians certified in family medicine between 1989 and 1991 and residing in Ontario in 1993 and to gather information on the scope of practice of family physicians in the cohort in 1999. DESIGN: Responses to a mailed questionnaire sent in 1999 were compared with responses to a 1993 survey of this group. SETTING AND PARTICIPANTS: All family physicians in Ontario in 1993 who received certification in 1989, 1990, or 1991 after completing a family medicine residency. Seven of 557 respondents to the 1993 survey were ineligible; 293 physicians (53%) responded to the 1999 survey. MAIN OUTCOME MEASURES: Field, location, and scope of practice. RESULTS: About 91% of the cohort were still practising family medicine, although 11% of these had restricted their practices to certain areas within family medicine. Physicians migrated from Ontario (6%) in nearly equal numbers to other provinces and other countries, predominantly the United States. More family physicians offered counseling, shared antenatal care, and newborn care in 1999 than in 1993. Those with restricted family practices provided fewer types of services and were less likely to provide antenatal or intrapartum care or to provide in-hospital services. CONCLUSION: Receiving certification in family medicine does not guarantee that physicians will remain in family practice 8 to 10 years later. Loss from general family medicine to restricted practices within family medicine and specialization was greater than loss from migration. PMID- 11494929 TI - Professional activity. How is family physicians' work time changing? AB - OBJECTIVE: To examine hours worked professionally, work preferences, and changes in both of these and their correlates. DESIGN: Repeated surveys done in 1993 and 1999. SETTING: Ontario family practices. PARTICIPANTS: Cohort of physicians certified in family medicine between 1989 and 1991 after family medicine residency who were surveyed in 1993 when they resided in Ontario. MAIN OUTCOME MEASURES: Self-reported hours spent weekly on professional activities, desired hours of professional work, and balance between work and other activities. RESULTS: Fifty-three percent (293) of 553 physicians responded to the 1999 survey; 91% had remained family physicians; 85% of these had participated in the 1993 survey. The difference between the hours that family physicians preferred to work professionally and their actual hours of work had increased since 1993. Childless physicians, women physicians with preschool children, and women physicians married to other physicians worked fewer hours professionally than other physicians in 1999. Female physicians and physicians without children worked closer to their preferred hours than other physicians. Reporting a preference to work fewer hours professionally in 1993 was linked with a reduction in professional activities by 1999. CONCLUSION: Greater attention should be paid in physician resource planning to the family life cycle of female physicians. Lifestyle changes could lead to a reduction in professional activity among these physicians. PMID- 11494930 TI - Pap test results. Responding to Bethesda system reports. AB - OBJECTIVE: To review the adequacy and diagnostic categories of the Bethesda system for reporting Pap test results (cervicovaginal cytology) and summarize management options. QUALITY OF EVIDENCE: The latest research evidence and guidelines from both international and Canadian sources are reviewed. With a few exceptions, good evidence supports particular management approaches for each adequacy statement and diagnostic category. MAIN MESSAGE: Women with unsatisfactory Pap smears should be re-examined and retested. Women with satisfactory smears and a diagnosis of "within normal limits" (WNL) or "benign cellular changes" (BCC) should be retested only at recommended screening intervals. Women with "satisfactory but limited by..." results and a diagnosis of WNL or BCC should have individualized follow up. Women with diagnoses of high grade squamous intraepithelial lesions, atypical glandular cells of uncertain significance, or malignancy should have further investigation (colposcopy). Optimal management of asymptomatic women with normal cervices and reports of atypical squamous cells of uncertain significance or low-grade squamous intraepithelial lesions is still controversial. CONCLUSION: Management of women following Pap tests is determined by both the adequacy of the test and diagnoses based on the results. PMID- 11494931 TI - Case report: unexplained syncope explained. PMID- 11494932 TI - Hypothesis: the research page. Part 3: Power, sample size, and clinical significance. PMID- 11494933 TI - Organizing the approach to musculoskeletal misuse syndrome. AB - Musculoskeletal misuse syndromes are secondary to misuse of specific tendons, ligaments, or other soft tissues. Despite patient education interventions to prevent acute musculoskeletal injury, these injuries still occur. A well organized approach to musculoskeletal complaints can reduce the risk of missed diagnoses and ensure appropriate care. This article reviews history and physical examination findings indicative of a misuse etiology and provides clinicians with a discussion of available diagnostic tests and therapies. PMID- 11494934 TI - Assessing the female sexual assault survivor. AB - Because sexual assault survivors often seek treatment in primary care settings, clinicians must be prepared to evaluate these patients in a nonjudgmental manner. The initial evaluation includes a medical and assault history. During the physical examination, physical injuries are noted and forensic evidence is collected. Treatment includes prophylaxis for pregnancy and sexually transmitted infections and counseling referrals. This article focuses on care of the adult female sexual assault survivor. Psychosocial support, physical examination, collection of evidence, treatment, and documentation are discussed. PMID- 11494935 TI - Antidepressant prescribing practices of nurse practitioners. AB - Depression continues to be seriously underdiagnosed and undertreated in primary care settings. Many studies have observed antidepressant prescribing practices of nonpsychiatric physicians to discover why inadequate depression detection and treatment is pervasive. This article describes the first large, randomized study to investigate the antidepressant prescribing practices of NPs. This quantitative, descriptive study sought to determine the factors NPs consider when choosing an antidepressant, antidepressant prescribing frequency by NPs, types of antidepressants prescribed by NPs, and health problems that NPs are treating with antidepressants. The study's findings indicate that NPs should adhere more closely to established practice guidelines. PMID- 11494936 TI - An adolescent with a polymorphous rash. PMID- 11494937 TI - Seeing it for yourself. PMID- 11494938 TI - TOMAC: an orthognathic treatment planning system. Part 1 soft-tissue analysis. PMID- 11494939 TI - Choosing the right partner. PMID- 11494940 TI - Orthodontic traction of an impacted maxillary central incisor. PMID- 11494941 TI - Accelerated Invisalign treatment. PMID- 11494942 TI - Digital models: a new diagnostic tool. PMID- 11494943 TI - Retrofitting curing lights for high-speed bonding. PMID- 11494944 TI - Bonding interdental retainers in patients with generalized marginal bone loss. PMID- 11494945 TI - Predictable success with resin bonded fixed partial dentures. AB - The conservative nature of the resin bonded fixed partial denture is the reason it is used. The ability to replace a missing tooth while avoiding aggressive preparation of the abutment teeth is a desirable concept. The use of conservative preparation forms will aid in the long term predictable success of the resin bonded fixed partial denture. PMID- 11494947 TI - The magic tooth' gives children the mouthful they deserve. PMID- 11494946 TI - Non-odontogenic toothache. PMID- 11494948 TI - Special Merit Award for professor John J. Clarkson. PMID- 11494949 TI - Report on the SCAW conference on pain, distress and stress in research animals. Current Standards and IACUC responsibility. May 18-19, 2000, Baltimore, MD. PMID- 11494951 TI - Primer on correlation coefficients. PMID- 11494952 TI - Familial hypertrigliceridemia. PMID- 11494953 TI - Clinical quiz: Henoch-Schonlein-like leukocytoclastic vasculitis. PMID- 11494955 TI - Managing aggressive thoughts and feelings with daily counts of non-aggressive thoughts and feelings: a self-experiment. AB - This article describes a self-experiment with a participant managing aggressive thoughts and feelings. The participant counted occurrences of aggressive thoughts and feelings per 24 h, and displayed these data on Standard Celeration Charts. Our experimental questions addressed the effects of daily 1-minute counts of non aggressive thoughts and feelings, and daily distributed series of six 10-second counts of non-aggressive thoughts and feelings on the occurrences of aggressive thoughts and feelings. We used an A1-B1-C1-B2-C2-A2 experimental design to analyze data from the 1-minute counts and the six 10-second distributed counts as these conditions alternated around two baseline conditions. Compared to the baseline, less aggressive thoughts and feelings occurred during both independent variable conditions. The six 10-second distributed counts produced lower frequencies of aggressive thoughts and feelings than the 1-minute counts. At the end of the second six 10-second counting procedure and during the second baseline, the participant most frequently had 0 aggressive thoughts and feelings per day. PMID- 11494954 TI - Social phobia viewed as a problem in social functioning: a pilot study of group behavioral treatment. AB - Available treatments of social phobia reduce anxiety and avoidance of social situations. Social functioning however, remains relatively unaffected. This study piloted a behavioral group treatment that aimed at improving patients' individual difficulties in social functioning. Five patients fulfilling DSM-IV criteria for social phobia were selected for treatment by a 16-session behavioral group treatment focusing on improving interpersonal functioning both in specific phobic situations as well as in different spheres of social life in general. Four out of 5 patients completed treatment and showed individual patterns of clinically significant improvement. Their individual test-score results and case-histories are presented. Three patients no longer fulfilled to the diagnostic criteria at the end of treatment; the fourth--at 6 month follow-up. This was maintained at a 2 year follow-up. PMID- 11494956 TI - A controlled study into the (cognitive) effects of exposure treatment on trauma therapists. AB - Several researchers have reported on therapists' symptoms as a result of trauma treatment, such as disruptions in cognitive schemata and symptoms resembling PTSD symptoms. Thus far, however, no studies compared the symptoms of trauma therapists and non-trauma therapists. In the present study, both trauma therapists (n = 20) and non-trauma therapists (n = 19) were included. During semi structured interviews, both therapist groups reported negative and positive effects of their work with patients on their personal and professional functioning. Trauma therapists reported more often changes of cognitions due to trauma work. Results from questionnaires, however, showed that compared with norm standards and compared with non-trauma therapists, trauma therapists did not show enhanced psychopathology nor distorted cognitive schemata. It is concluded that although exposure treatments with trauma patients are associated with therapists' distress, therapists seem able to cope with it in a healthy way. PMID- 11494957 TI - Assessing the perceived predictability of anxiety-related events: a report on the perceived predictability index. AB - Predictability, or lack thereof, is believed to play a critical role in the development and maintenance of anxiety, with unpredictability being associated with heightened levels of anxious and fearful responding. Despite the potential importance of predictability in theoretical accounts of emotional dysregulation, currently no standardized assessment instrument exists to assess predictability perceptions for anxiety-related events. The present series of four investigations report on an initial attempt to develop a self-report instrument (i.e., Perceived Predictability Index, PPI) that can measure predictability perceptions for the occurrence, duration, and termination of anxiety-related events. Initial item selection and factor structure of the instrument was based on a large sample of participants and yielded a two-factor solution: (1) prediction of anxiety-related environmental events and (2) prediction of internal events. Our subsequent studies show that the PPI possesses adequate levels of internal consistency and temporal stability over time. Additionally, the PPI demonstrated adequate divergent and convergent validity relative to other standard anxiety and fear measures. The internal dimension of the scale also demonstrated predictive validity for emotional responding during a biological challenge test. We discuss these findings in relation to the role of perceived predictability in the study of anxious and fearful responding, and offer directions for future research. PMID- 11494958 TI - Pharmacotherapy for severe aggression in a child with autism: "open label" evaluation of multiple medications on response frequency and intensity of behavioral intervention. AB - Many persons who have developmental disabilities and challenging behaviors are treated with multiple medications combined with nonpharmacological approaches. However, the comparative effects from pharmacotherapy frequently are not assessed empirically, do not include corollary behavioral measures, and are not evaluated in the long term. The present single-case study incorporated behavioral assessment methodology in an "open label" evaluation of anticonvulsant (clonazepam), beta-blocking (propanolol), and antidepressant (sertraline and clomipramine) medications on severe aggression in a child with autism. Clinically significant reductions in aggressive behavior were attained with the administration of clomipramine and the reductive effects from the medication persisted for 1.7 years. In addition, clomipramine was associated with the elimination of crisis intervention procedures that had been required to manage the child's aggression. These findings add to the clinical literature describing effective treatment of serious behavior disorders in persons with developmental disabilities using antidepressant medication. PMID- 11494959 TI - Investigating the effects of physical discomfort on laboratory-induced mood states. AB - In order to investigate the effects of physical discomfort (i.e., pain) on the induction of mood, 139 participants were randomly assigned to one of four conditions: sad, sad with physical discomfort, happy, and happy with physical discomfort. Consistent with our hypotheses, the addition of physical discomfort resulted in a significantly stronger induction of self-reported depression, with these findings showing temporal specificity and persisting even after statistically controlling for stable response biases. However, other self reported affective states, as well as cognitive and behavioral measures, were not significantly affected by physical discomfort. Findings are discussed with regard to making the laboratory induction of sad affect more consistent with the multifaceted experience of depression. PMID- 11494960 TI - Efficacy of virtual reality exposure therapy to treat driving phobia: a case report. AB - An AB case design was used to examine the efficacy of virtual reality exposure therapy (VRET) in treating driving phobia. After a one week baseline, the patient received three treatment sessions over a ten day period. Treatment included practice of four VR driving scenarios. Peak anxiety decreased within and across sessions. Ratings of anxiety and avoidance declined from pre-treatment and post treatment, with gains maintained at seven month followup. Phobia-related interference in daily functioning similarly decreased. The results suggest that it would be useful to further evaluate the efficacy of VRET for driving phobia in controlled clinical trials. PMID- 11494961 TI - Imaginal exposure for anger reduction in adult outpatients: a pilot study. AB - Although exposure procedures have been widely accepted in the treatment of anxiety disorders, they have rarely been applied to the treatment of anger. The present paper describes an initial attempt to apply an imaginal exposure strategy to adult outpatients (n = 6) referred for anger management. This investigation reflects an empirical clinical practice approach rather than a controlled outcome study. Thus, this paper provides a clinical description of the imaginal exposure program, pre-to-posttest effectiveness data, an exploration of habituation patterns for each participant, and 15-month follow-up data from several patients. In considering the impact of the intervention, statistically significant change was found on most anger variables, the majority of patients met a criteria for clinically significant improvement on important indices of anger, and treatment effect sizes were large and compared favorably to previously studied interventions. Process data revealed a consistent habituation effect, across patients and anger stimuli, in response to repeated exposure practice. Participants' satisfaction was also positive. Finally, statistically significant and clinically meaningful change was evident at 15-months following the intervention. Data from the current pilot project are encouraging and hopefully will stimulate more methodologically rigorous clinical trials. PMID- 11494962 TI - Dutch patients with glycogen storage disease type II show common ancestry for the 525delT and del exon 18 mutations. PMID- 11494963 TI - Recurrent mutations in the deafness gene GJB2 (connexin 26) in British Asian families. PMID- 11494964 TI - A region of homozygosity within 22q11.2 associated with congenital heart disease: recessive DiGeorge/velocardiofacial syndrome? PMID- 11494965 TI - High frequency of the ApoB-100 R3500Q mutation in Bulgarian hypercholesterolaemic subjects. PMID- 11494966 TI - A community based study on intentions regarding predictive testing for hereditary breast cancer. PMID- 11494967 TI - Ocular malformations, postaxial polydactyly, and delayed intramembranous ossification: a new autosomal dominant condition. PMID- 11494968 TI - Recessively inherited lower incisor hypodontia. PMID- 11494969 TI - Congenital diaphragmatic hernia and interstitial deletion of chromosome 3. PMID- 11494970 TI - A supernumerary marker chromosome with a neocentromere derived from 5p14- >pter. PMID- 11494971 TI - A case of Roberts syndrome described in 1737. PMID- 11494974 TI - Image of the month: hepatitis C. PMID- 11494972 TI - Global health fund and global realities. PMID- 11494975 TI - Lichenoid eruptions due to ursodeoxycholic acid administration. PMID- 11494976 TI - Ileorectal anastomosis in patients with familial adenomatous polyposis. PMID- 11494977 TI - Ileorectal anastomosis is appropriate for a subset of patients with familial adenomatous polyposis. PMID- 11494978 TI - Osteoporosis in newly diagnosed inflammatory bowel disease. PMID- 11494979 TI - Acute-phase protein, survival and tumour recurrence in patients with colorectal cancer (Br J Surg 2001; 88: 255-60). PMID- 11494980 TI - Acute-phase protein, survival and tumour recurrence in patients with colorectal cancer (Br J Surg 2001; 88: 255-60). PMID- 11494981 TI - Value and cost of follow-up after adjuvant treatment of patients with Dukes' C colonic cancer (Br J Surg 2001; 88: 101-6). PMID- 11494982 TI - Randomized clinical trial of diathermy versus scalpel incision in elective midline laparotomy (Br J Surg 2001; 88: 41-4). PMID- 11494983 TI - Evaluation of the necessity for gastrectomy with lymph node dissection for patients with submucosal invasive gastric cancer (Br J Surg 2001; 88: 444-9). PMID- 11494984 TI - Randomized clinical trial of Helicobacter pylori from dental plaque(Br J Surg 2001; 88: 200-5). PMID- 11494985 TI - pH dependence and solvent deuterium oxide kinetic isotope effects on Bacillus cereus beta-lactamase I catalyzed reactions. AB - The solvent kinetic isotope effects (SKIE's) on k(cat) (D(V)) and on k(cat/Km[D(V/K)] were determined for the Bacillus cereus beta-lactamase I catalyzed hydrolysis of five substrates that have values of k(cat)/K(m) varying over the range (0.014-46.3) X 10(6)M(-1) s(-1) and of k(cat) between 0.5 and 2019 s(-1). The variation of D(V/K) was only from 1.06 to 1.25 among these compounds and that in D(V) was from 1.50 to 2.16. These results require that Dk(1), the SKIE on the enzyme-substrate association rate constant, and D(k-1/k2), that on the partition ratio of the ES complex, both be near 1. The larger SKIE observed on D(V) requires that an exchangeable proton be in flight for either or both the acylation and the deacylation reaction. The pH dependence of the values k(cat)/K(m) for three substrates shows identical pK(a)s of 5.5. and 8.4. This identity combined with the fact that only one of these three substrates is kinetically "sticky" proves that the substrates can combine productively with only one protonic form of the enzyme. There is considerable substrate variation in the pK(a) values of k(cat) observed vs. pH profiles; the inflection points for all substrates studied are at pH values more extreme than are observed in the pH profiles for k(cat)/K(m). PMID- 11494986 TI - Dirty questions: Indigenous health and 'Western research'. AB - OBJECTIVE: This paper explores both Indigenous and non-indigenous critiques of 'Western' research frameworks in an Aboriginal health context. It also discusses the 'reform' of Aboriginal health research practices since the 1980s, particularly in relation to the development of ethical guidelines. METHOD: The text is based on both archival research and a critical review of secondary literature. CONCLUSIONS AND IMPLICATIONS: It is argued here that efforts to reform the practices of mainstream Indigenous health research since the 1980s have oscillated between taking concrete steps towards actually changing research practice and placing too great a reliance on written guidelines and positive rhetoric. In offering this analysis, the paper argues for a more challenging conception of reforming mainstream research, involving an emphasis on shifts in institutional arrangements as well local research practices. PMID- 11494987 TI - Passive smoking and lung cancer: a cumulative meta-analysis. AB - OBJECTIVE: To review the epidemiological evidence for the association between passive smoking and lung cancer. METHOD: Primary studies and meta-analyses examining the relationship between passive smoking and lung cancer were identified through a computerised literature search of Medline and Embase, secondary references, and experts in the field of passive smoking. Primary studies meeting the inclusion criteria were meta-analysed. RESULTS: From 1981 to the end of 1999 there have been 76 primary epidemiological studies of passive smoking and lung cancer, and 20 meta-analyses. There were 43 primary studies that met the inclusion criteria for this meta-analysis; more studies than previous assessments. The pooled relative risk (RR) for never-smoking women exposed to environmental tobacco smoke (ETS) from spouses, compared with unexposed never smoking women was 1.29 (95% CI 1.17-1.43). Sequential cumulative meta-analysed results for each year from 1981 were calculated: since 1992 the RR has been greater than 1.25. For Western industrialised countries the RR for never-smoking women exposed to ETS compared with unexposed never-smoking women, was 1.21 (95% CI 1.10-1.33). Previously published international spousal meta-analyses have all produced statistically significant RRs greater than 1.17. CONCLUSIONS: The abundance of evidence in this paper, and the consistency of findings across domestic and workplace primary studies, dosimetric extrapolations and meta analyses, clearly indicates that non-smokers exposed to ETS are at increased risk of lung cancer. IMPLICATIONS: The recommended public health policy is for a total ban on smoking in enclosed public places and work sites. PMID- 11494988 TI - Potentially avoidable hospitalisations in New Zealand, 1989-98. AB - OBJECTIVE: To describe potentially avoidable hospitalisation in New Zealand, including recent trends and variations between groups differentiated by age, gender, ethnicity and degree of deprivation. METHOD: Hospital discharges among people aged 0-74 years for the years 1989-98 were classified as 'potentially avoidable' or 'unavoidable' based on the ICD9-CMA code of the principal diagnosis. Potentially avoidable hospitalisations (PAH) were further subcategorised according to the intervention involved--primary prevention, ambulatory care or injury prevention. RESULTS: By 1998, one in three of these hospitalisations was theoretically avoidable--two-thirds of these through more effective primary health care services. Although in practice only a proportion of these could realistically have been avoided, these estimates reveal considerable scope for further reduction in the incidence of serious disease and injury. Maori and Pacific people had age-standardised PAH rates approximately 60% higher than European and other New Zealanders. Similar discrepancies exist by socio-economic deprivation. Had all New Zealanders enjoyed the PAH rates of the most advantaged 40% of the population, 28% fewer potentially avoidable hospitalisations would have occurred in 1998, some 26,000 hospital admissions. CONCLUSION: This analysis has revealed significant scope for the health sector to contribute to population health gain and, in particular, to improvement in equity of outcomes across ethnic and socio-economic groups. Potentially avoidable hospitalisations provide a useful tool for evidence-based population health needs analysis and health policy development. PMID- 11494989 TI - An economic evaluation of universal infant vaccination against hepatitis B virus using a combination vaccine (Hib-HepB): a decision analytic approach to cost effectiveness. AB - OBJECTIVE: To evaluate the health impact and cost effectiveness of two infant vaccination strategies for protection against hepatitis B virus (HBV) infection in the Australian population. Vaccinating only high-risk infants, assuming 65% compliance, was compared with universal vaccination of infants using a combination Hib-HepB vaccine, with 87.4% compliance. METHOD: A Markov model simulated the natural history of HBV infection and disease in an Australian birth cohort. The cohort was divided into those at high risk of infection (infants born into high-risk families) and low-risk infants. Clinical and epidemiological data used were obtained from published reports and a survey of clinical experts. The model included the health costs associated with acute and chronic HBV infection, and the sequelae of chronic HBV infection. RESULTS: The model predicted that universal hepatitis B vaccination of an Australian birth cohort (260,000 births) would result in a 77% reduction in cases of HBV infection. The incremental cost per life year gained was $11,862, which is low compared with many other health care interventions. With no discounting of costs or consequences, universal vaccination with the combination vaccine was predicted to save lives and reduce costs. CONCLUSION: There is no socially accepted threshold value for cost per life year gained to guide decisions about funding Australian health care interventions. Nevertheless, based on these results, universal hepatitis B vaccination of Australian infants using a combination Hib-HepB vaccine would almost certainly be regarded as a worthwhile investment of public funds. PMID- 11494990 TI - Adolescent hepatitis B immunisation--should it be the law? AB - Universal hepatitis B immunisation of young adolescents was included in the Australian Standard Vaccination Schedule in 1998. However, rates of immunisation among adolescents world-wide have often been inadequate. Australia's experience in this area is no exception, particularly in States where school-based delivery is not carried out. Legislation for pre-school immunisation certification currently exists in several States and this legislation is distinctly different from the compulsory or mandatory immunisation that exists in several other developed countries. There have been demonstrable gains in uptake as a result of mandatory immunisation requirements in the United States and there is evidence to suggest that immunisation certification in Australia has also been beneficial. However, it is important to recognise that both certification and mandatory immunisation legislation have inherent difficulties. In this paper, we argue that legislation for high school immunisation certification, as part of a multi faceted vaccine delivery strategy tailored to adolescents, is required to achieve the uptake that will lead to interruption of transmission of the hepatitis B virus in Australia. Not only will it substantially reduce incident cases of hepatitis B for the next decade, it will also provide a framework for the successful introduction of future adolescent vaccine initiatives in Australia. PMID- 11494991 TI - Raw oyster consumption and outbreaks of viral gastroenteritis in New Zealand: evidence for risk to the public's health. AB - OBJECTIVE: To investigate the relationship between consumption of raw Pacific half shell oysters and outbreaks of Norwalk-like virus (NLV) gastroenteritis in Auckland in the last third of 1999. METHOD: Ten outbreaks were investigated as retrospective cohorts using standardised questionnaires relating to food and drink exposures. Trace back of oysters and site inspections of implicated commercial growing areas were performed. Virological analyses compared oysters linked to outbreaks and faecal samples from cases. RESULTS: Eighty-six cases were identified, of whom 32 (37.2%) were confirmed NLV positive on faecal analysis. The summary risk estimate for illness among oyster consumers for all outbreaks was RR 8.23 (95% CI 4.55-14.90; p< 0.001) and in five of seven outbreaks permitting statistical analysis, the risk for those consuming raw oysters was greater than five-fold that of non-consumers. NLVs were identified in two batches of oysters from different growing areas, implicated in four outbreaks. In both the strain (Genogroup II/3 'Mexico-like virus') from cases matched that in the oysters from the same harvest batch. CONCLUSION: The epidemiological and virological evidence implicates oysters as the source of a number of outbreaks of NLV gastroenteritis. This is the first time NLVs have been identified in commercially farmed Pacific oysters in New Zealand. Sewage effluent from recreational boats was the likely source of faecal contamination of growing waters in one site. IMPLICATIONS: Combined use of virological and epidemiological methods have proved invaluable in investigating NLV outbreaks. Further research is needed into enteric viral contamination of commercial oyster farms. PMID- 11494992 TI - Investigation of potential iatrogenic transmission of hepatitis C in Victoria, Australia. AB - OBJECTIVE: To determine the level of exposure to medical and surgical procedures among Australian-born patients whose mode of acquisition of the hepatitis C virus (HCV) is unknown. METHOD: Place and time of study: Melbourne, Australia, 1998 2000. DESIGN: Retrospective case series. INSTRUMENT: Structured questionnaire administered by one interviewer. SETTING: Referral centre for hepatitis C in a tertiary teaching hospital. PARTICIPANTS: Australian-born individuals persistently HCV antibody (anti-HCV) positive on at least two second-generation commercial assays. MAIN OUTCOME MEASURES: Demographic and self-reported exposure data. RESULTS: Of 135 anti-HCV positive individuals with no known mode of transmission, 54 (40%) individuals fulfilled all the entry criteria and agreed to participate. Of the 54 cases, 53 had at least one medical/surgical procedure and/or invasive dental work; 46 (85%) had dental extractions, 19 (35.2%) had complex dental work, e.g. root canal, 44 (82%) had an operation requiring general anaesthesia, 41 (75.9%) had a procedure requiring local anaesthetic, and a number of endoscopic procedures were reported: gastroscopy (n=3), colonoscopy (n=3), laparoscopy (n=4), arthroscopy (n=5), cystoscopy (n=2). CONCLUSION: We have documented exposure to medica/surgical procedures among HCV patients with no previously recognised mode of transmission. IMPLICATIONS: The findings of this study have important public health implications for current cleaning, disinfection and sterilisation procedures and protocols (or lack of these) as well as for the policies and guidelines relating to the re-use of medical equipment such as multi-dose vials, suturing material and anaesthetic circuits. PMID- 11494993 TI - Behavioural factors as predictors of motor vehicle crashes: differentials between young urban and rural drivers. AB - OBJECTIVE: To describe the driver and behavioural factors that predispose young drivers to crash in the first 12 months of driving and to compare whether these factors differ between young urban and rural drivers. METHODS: A cohort comprising 1,796 newly licensed urban and rural drivers from Western Australia was recruited and followed over the first 12 months of driving. Using Cox proportional hazard analysis, driver and behavioural factors were assessed to determine whether they predicted the likelihood of a crash. RESULTS: The incidence rate for a motor vehicle crash was marginally higher for urban drivers compared with rural drivers (Urban: IR=4.2/10,000 driving days; Rural: IR=3.7/10,000 driving days). There was no significant difference in the time to crash between urban and rural drivers. Two factors, namely the frequency of driving before obtaining a learner-driver permit and the driver's level of risk taking, were significantly associated with a motor vehicle crash in the first year of driving. CONCLUSION: Irrespective of whether the driver is licensed in a rural or urban area, high risk-taking drivers are at an elevated risk of a motor vehicle crash in the first 12 months of driving. IMPLICATIONS: Aspects of Graduated Driver Training and Licensing Programs (GDTLP) could be effective in targeting this at-risk group. PMID- 11494994 TI - Increasing mouthguards usage among junior rugby and basketball players. AB - OBJECTIVE: To evaluate a Western Australian mouthguard promotion campaign, launched at the start of the 1997/98 junior rugby union and junior basketball seasons, aimed at increasing mouthguard usage at competition and training. METHOD: A quasi-experimental field design was used to assess the impact of the mouthguard campaign on behavioural change. Observational data were collected pre- and post-campaign on mouthguard usage by players present at a rugby and basketball competition event and at a training session. Junior Australian Rules Football players were used as a control group. RESULTS: Pre-post observational surveys showed a significantly greater increase in mouthguard usage in competition games among rugby union (77% to 84%) and basketball players (23% to 43%) compared with the control group (72% to 73%). All codes showed a post campaign increase in mouthguard usage at training, but the intervention codes' increases were greater than the control's increase (rugby union: 29% to 40%; basketball: 11% to 36%; football: 34% to 40%). CONCLUSIONS: The campaign had a significant and substantial effect on behaviour and provides evidence of the benefits of leveraging a sponsorship to modify the behaviour of the target group. IMPLICATIONS: This campaign provides a model for promoting mouthguard usage in other sports among junior players. PMID- 11494995 TI - Compliance of Dunedin school playground equipment with the New Zealand playground standard. AB - OBJECTIVE: Injuries resulting from falls from playground equipment are a public health concern in New Zealand. Like many other countries, New Zealand has a safety standard aimed at reducing the incidence and severity of these injuries by limiting the height from which children can fall from playground equipment and requiring the provision of impact-absorbing surfaces beneath equipment from which falls are possible. The purpose of this study was to examine progress towards achieving compliance with these requirements in Dunedin school playgrounds. METHODS: Sixty-two schools were audited over the summer of 1997/98 and information recorded on equipment type, maximum fall height, surface type, and depth of loose-fill surface materials. Comparisons were made with audits conducted in 1989 and 1981. RESULTS: Substantial increases in the amount of playground equipment and in the provision of impact-absorbing surfaces were observed. A small increase in compliance with the requirement that the maximum fall height of equipment not exceed 2.5 metres was also observed. CONCLUSIONS: Any gains in safety achieved through increased compliance with the height and surface requirements of the New Zealand Standard have been counteracted by the substantial increase in the amount of equipment available in playgrounds. IMPLICATIONS: A more drastic measure is needed to achieve a meaningful reduction in the incidence of injury following falls from playground equipment. PMID- 11494996 TI - Cervical screening by socio-economic status in Australia. AB - OBJECTIVES: To examine differentials and time trends in self-reported Pap test rates by socio-economic status (SES) from the 1989/90 and 1995 Australian National Health Surveys (NHS). METHODS: The unit record data for females were extracted from the two NHSs and combined. The outcome variable of interest was 'having a Pap test in the past three years'. The principal study factor was SES measured as individual characteristics and SES of area of residence. Migrant status, rurality, year of survey and age were controlled for in logistic regression models. RESULTS: Self-reported rates of having a Pap test in the past three years were higher in women from higher compared with lower SES groups. Compared with women with a bachelor or higher degree, the odds of reporting having a Pap test in the past three years in women with no post-school qualification was 0.86 (p<0.0005). Women with a gross annual income of less than $20,000 had significantly lower odds (OR=0.79) compared with women earning $40,000 or more. Blue collar (OR=0.84) and not employed (OR=0.73) women also had significantly lower odds compared to the referent white collar group. CONCLUSION: This study reveals differentials in Pap screening behaviour by individual measures of SES in Australia. Area-based SES measures under-estimated the SES differentials in Pap test rates compared with individual measures. Derived population attributable fractions reveal that about a quarter of self-reported under-screening is accounted for by low SES when measured individually, compared to 8% when SES is measured ecologically. PMID- 11494997 TI - Long-term care of people below age 65 with severe acquired brain injury: appropriateness of aged care facilities. AB - OBJECTIVE: To identify the number of people younger than 65 years with acquired brain injury (ABI) living in aged care facilities in Queensland, and to evaluate the appropriateness of this accommodation option. METHODS: A cross-sectional descriptive study of all 493 Commonwealth Department of Health and Aged Care registered aged care facilities in Queensland. Associations between a range of demographic factors, resources, care provision and client needs were examined, from the perspective of service providers. RESULTS: The response rate was 75%. Twenty-six per cent of facilities (n=97) were providing care for 209 people younger than 65 years with ABI. The social, cognitive and rehabilitation aspects of client care were found to be inadequate in facilities where staffing levels, training and funding resources were limited (p< 0.05). Smaller facilities (<60 beds) reported higher levels of family participation in specific aspects of client care (p<0.05). Almost 40% of the facilities indicated they did not adequately meet the specific and complex rehabilitation needs of these clients. Aged care facilities were the least favoured model of care for this client group (8%) compared with the most favoured model of small group homes (46%). CONCLUSIONS: The current use of aged care facilities for housing younger people with high-level care needs resulting from ABI is inappropriate and does not meet client needs. PMID- 11494998 TI - John's $12 tonic: press coverage of the government's selling of a private health insurance rebate. AB - OBJECTIVE: To document representations of the 1998 introduction of a 30% rebate on private health insurance in the three most-read daily Sydney newspapers. METHODS: Thematic frame analysis of 131 newspaper articles. RESULTS: The rebate was opposed through two frames: that it was ineffective and unfair, and that it was politically motivated. Four supportive frames were more complex: the rebate was justified by claims that public health care was collapsing, that responsible citizens should pay for their own health care, and that individuals would benefit financially. There was also a focus on the political battle in the Senate. The newspaper with the readership least likely to benefit from the rebate supported it most strongly. CONCLUSIONS: Framing was strongly episodic (two dimensional, decontextualised and case-study based), limiting political accountability, and the anti-rebate case was presented less memorably. Community action around the issue was not encouraged, individual responsibility was emphasised and universal health care was not promoted as fair or necessary. Different readerships received different messages about the rebate. IMPLICATIONS: There is an urgent need to promote the value of the public health care system and make the future of Medicare compelling for news editors and the public. PMID- 11494999 TI - The Second Computer Assisted Telephone Interview (CATI) Forum: the state of play of CATI survey methods in Australia. AB - Computer Assisted Telephone Interview (CATI) systems are recognised in Australia and internationally for their ability to provide timely and relevant data on the health of the population. The Second CATI Population Health Surveys Forum highlighted the importance of this system for surveillance and using this information to determine priority health issues, develop strategies, monitor effectiveness of interventions, and influence health policy. A national CATI data collection strategy is required to determine priorities for health surveillance. Several development and technical issues need to be addressed for such a national strategy to provide a coordinated approach to health surveillance. PMID- 11495000 TI - Public support for smoking bans in bars and gaming venues. PMID- 11495001 TI - Transmission of HCV in the Vietnamese community. PMID- 11495002 TI - Rantes distribution and cellular localization in the brain of HIV-infected patients. AB - OBJECTIVE: To study the immunochemical distribution ofRantes chemokine and its correlation with HIV-p24 expression, in brains with HIV-related lesions. MATERIAL AND METHODS: 17 HIV-positive cases of HIV-related brain lesions, 7 HIV-positive cases without cerebral HIV-related lesions (5 with opportunistic brain diseases), and 7 HIV-negative cases as controls (4 with brain lesion) were selected. RESULTS: High expression of Rantes was observed in the cases with inflammatory brain lesions (22/24 HIV-positive and 2/7 HIV-negative patients). Positivity was observed in the diffuse and nodular microglial cells and lymphocytes. In the patients with HIV-related lesions, the presence of Rantes-stained microglia did not correlate with that of HIV-p24-positive cells. Positive astrocytes were only found in the HIV-positive patients. Multinucleated giant cells were always Rantes negative. CONCLUSIONS: Our results seem to demonstrate the role of Rantes chemokine in inducing inflammatory brain perivascular and microglial reactions both in HIV-positive and -negative patients. PMID- 11495003 TI - Neuropathology and neurodegeneration in human immunodeficiency virus infection. Pathogenesis of HIV-induced lesions of the brain, correlations with HIV associated disorders and modifications according to treatments. AB - A variety of HIV-induced lesions of the central nervous system (CNS) have been described, including HIV encephalitis, HIV leukoencephalopathy, axonal damage, and diffuse poliodystrophy with neuronal loss of variable severity resulting, at least partly, from an apoptotic process. However, no correlation could be established between these changes and HIV dementia (HIVD). From our study of HIV infected patients, it appeared that neuronal apoptosis is probably not related to a single cause. Microglial and glial activation, directly or indirectly related to HIV infection, plays a major role in neuronal apoptosis possibly through the mediation of oxidative stress. In our patients with full-blown AIDS, this mechanism predominated in the basal ganglia and correlated well with HIVD. Axonal damage, either secondary to microglial activation, or to systemic factors also contributes to neuronal apoptosis. Although massive neuronal loss may be responsible for HIVD in occasional cases, we conclude that neuronal apoptosis is a late event and does not represent the main pathological substrate of HIVD. The dementia more likely reflects a specific neuronal dysfunction resulting from the combined effects of several mechanisms, some of which may be reversible. Introduction of highly active antiretroviral therapy dramatically improved patient survival, however, its impact on the incidence and course of HIVD remains debatable. In our series, the incidence of HIVE has dramatically decreased since the introduction of multitherapies, but a number of cases remain whose cognitive disorders persist, despite HAART. The poor CNS penetration of many antiretroviral agents is a possible explanation, but irreversible "burnt out" HIV-induced CNS changes may also be responsible. PMID- 11495004 TI - Glioblastoma with large numbers of eosinophilic hyaline droplets in neoplastic astrocytes. AB - We report a case of glioblastoma with unusual histological features arising in the left frontal lobe of a 79-year-old woman. On routine histological examinations of specimens obtained at the surgical resection, the tumor was consistent with glioblastoma, and intracytoplasmic inclusions of bright eosinophilic, round objects were found in a large number of neoplastic astrocytes. Pathological studies using histochemical/ immunohistochemical stainings and electron microscopy demonstrated that the inclusions were compatible with eosinophilic hyaline droplets (EHD), which are predominantly seen in pleomorphic xanthoastrocytoma and pilocytic astrocytoma. EHD-bearing cells were distributed throughout the tumor tissue and focally abundant (182/mm2). Most of those cells were negative for MIB- I immunostaining, although mitotic figures were rarely observed. Neither round granular body nor Rosenthal fibers were seen. Based on these neuropathological findings and a review of the literature, we concluded that this case was a very rare case of glioblastoma with numerous EHDs. The presence of numerous EHDs is considered a diagnostically helpful feature suggesting low-grade astrocytomas. The present case suggested that diagnostic application of numerous EHDs should be careful, particularly in small biopsy samples such as stereotactic biopsy. PMID- 11495005 TI - Prognostic relevance of intracytoplasmic cytokeratin pattern, hormone expression profile, and cell proliferation in pituitary adenomas of akromegalic patients. AB - Seventy-six pituitary adenomas of akromegalic patients were investigated to find out the prognostic relevance of the intracytoplasmic distribution of cytokeratins (CK), immunohistochemically defined hormone production profile, proliferative activity and clinical presentation. CK distribution, growth fraction (MIB1 index) and hormone production profile were analyzed by means of immunohistochemistry. Apoptotic activity was investigated by the TUNEL method. Two different CK distribution patterns were seen: a dot-like pattern in 29 cases (type 1 adenomas), and a perinuclear fibrillary pattern in 47 cases (type 2 adenomas). Type 2 adenomas showed more prominent coexpression of prolactin (p < 0.0001), luteotrophic hormone (p < 0.002), follicle-stimulating hormone (p < 0.005), thyroid-stimulating hormone (p < 0.0001), and alpha-subunit (p < 0.005), as compared to type 1 adenomas. The mean MIB1 index was significantly higher in type 1 vs. type 2 tumors (4.23%, range: 1.93% - 9.83% vs. 2.07%, range: 0.67% - 4.87%, p < 0.0001). Apoptotic activity was too low in both examined groups to be used for balancing of tumor cell turnover. Clinical analysis of patients with type 1 adenomas revealed female predominance, younger age, larger tumor size, and more frequently aggressive growth with higher incidence of suprasellar extension (p < 0.0001) and cavernous sinus infiltration (p < 0.0001), as well as larger proportions of re-operations and incomplete resections (34.5% vs. 8.51%). Additionally, the interval until re-operation was shorter in type 1 adenomas (mean: 16 months, range: 9 - 21 months vs. mean: 57 months, range: 18- 158 months). We conclude that classification of adenomas of akromegalic patients based on intracytoplasmic CK distribution, combined with examination of proliferative activity, and immunohistochemically defined hormone production profile, provides important prognostic information for the management of akromegalic patients. PMID- 11495006 TI - Polyglucosan bodies and temporal lobe epilepsy: an incidental finding or more? AB - This study reports on histological findings in the temporal lobe of a 36-year-old woman who underwent a right temporal lobectomy for pharmaco-resistant complex partial seizures. Since surgery, the patient has remained seizure-free. The patient had an established diagnosis of right temporal lobe epilepsy, based on video EEG recordings of seizures, MRI hippocampal atrophy, focal interictal hypometabolism on fluoro-desoxyglucose, hypofixation of Cl1-flumazenil in PET studies, and ictal intracerebral recordings. Biopsies were studied under light- and electron microscopy. Histology showed diffuse distribution of a large number of polyglucosan bodies (PBs) in the whole right temporal lobe white matter. PBs were mostly confined to the perivascular areas and in subpial zones rarely and were observed in the most superficial cortical layers. There was some neuronal loss, especially in opercular zone T , but no other histological lesion was found. Ultrastructurally, PBs were made of filamentous and amorphous material, and were found both in intra-astrocytic processes and in axons. The presence of numerous PBs in the temporal lobe of patients with refractory temporal lobe epilepsy has been reported in 3 patients in the literature. It raises the questions whether this histological abnormality could be related to the epileptogenic process as a cause or as a consequence. PMID- 11495008 TI - Nitric oxide scavenging modulates an experimental vasoplesia in-vitro. AB - Endogenous overproduction of nitric oxide (NO) is believed to be a primary cause of refractory hypotension in septic shock. Under this condition, effectiveness of vasopressors is diminished due to hyporeactivity of blood vessels, a condition termed as vasoplesia. Effective reduction of NO levels should alleviate the condition. In this study, we investigated whether NO scavenging could modulate the endotoxin mediated vasoplesia in-vitro. Further, we explored whether NO scavenging in combination with a moderate NO synthase (NOS) inhibition would also be effective in modulating NO mediated vasoplesia. Experimental vasoplesia was produced in-vitro by incubating isolated rat thoracic aortic rings with lipopolysaccharide (LPS). Vessel rings were then treated with N(omega)-nitro-L arginine methyl ester (L-NAME; a NOS inhibitor), human hemoglobin (Hb; a NO scavenger), or both L-NAME and Hb. Vascular reactivity was assessed by measuring vessel ring isometric tension changes to norepinephrine (NE) doses; the median effective doses (logEC50) of NE before and after each experimental treatment were compared. Following a 6-hour LPS treatment, vascular reactivity logEC50 values for NE were significantly increased compared with control vessel rings incubated without LPS. Treatment with either L-NAME alone or Hb alone significantly improved the vessel ring reactivity to NE. When both L-NAME and Hb were used concomitantly, vascular reactivity was also significantly improved. These results indicate that NO scavenging with Hb is as effective as NO synthesis inhibition with NAME in modulating the endotoxin induced vasoplesia. In conclusion, NO scavenging, alone or in combination with a moderate NOS inhibition, may render an alternative therapeutic approach to NOS synthesis inhibition in modulating the vasoplesia in septic shock. PMID- 11495007 TI - Diagnostic utility of polymerase chain reaction and immunohistochemical techniques for the laboratory diagnosis of intracranial tuberculoma. AB - In an attempt to establish a tuberculous etiology, polymerase chain reaction (PCR) and immunohistochemical (IHC) methods were undertaken in formalin-fixed paraffin sections of ten surgical specimens of intracranial tuberculoma. The control group included an equal number of intracranial fungal granuloma. Both PCR and IHC methods did not yield false-positive results in fungal granuloma. PCR was found to be less sensitive (60%) than IHC method (80%) in this study. IHC method definitely possesses several operational advantages over PCR and is more suited to laboratories in developing countries for establishing a tuberculous etiology particularly in those patients in whom the conventional bacteriological methods did not confirm the diagnosis of tuberculoma. PMID- 11495009 TI - Superoxide generation from human polymorphonuclear leukocytes by liposome encapsulated hemoglobin. AB - We investigated the interactions between liposome-encapsulated hemoglobin (Neo Red Cells: NRC) and human polymorphonuclear leukocytes as assessed by superoxide generation. NRC triggered superoxide generation from neutrophils in a dose dependent manner. Empty liposomes also induced superoxide production of neutrophils. Superoxide generation of neutrophils induced by phorbol myristate acetate (PMA) was delayed but intensified both by NRC and empty liposomes. The intensity of superoxide generation induced by NRC was smaller than that by the empty liposomes. As NRC contained superoxide dismutase (SOD) that was copurified with hemoglobin from red blood cells and its activity remained, SOD contained in NRC may partially eliminate superoxide. PMID- 11495010 TI - Cross-linked hemoglobin as a potential membrane for an artificial red blood cell. AB - A cross-linked hemoglobin membrane has been created with discerning permeability between dissolved hemoglobin and small molecules. Such a membrane could be used to enclose a sphere of hemoglobin solution thereby allowing the entire "cell" to transport oxygen. The hemoglobin membrane was cross-linked on a polycarbonate support; the mechanical support was necessary for diffusion experiments in this study and would not be used during any sphere preparation. A 30% methemoglobin solution in phosphate buffer was used to fill the pores of the 10 microm polycarbonate support, then cross-linked with a homobifunctional cross-linking agent. The cross-linked hemoglobin within the support was evaluated for hemoglobin and benzoic acid permeability in a side-by-side diffusion cell. Disuccinimidyl glutarate, disuccinimidyl suberate and disuccinimidyl tartrate were used as cross-linking agents. Disuccinimidyl glutarate, 4.65 mM, created a hemoglobin-impermeable membrane after cross-linking for 20 minutes, reducing the concentration to 0.46 mM required a cross-linking time to 60 minutes. Benzoic acid, representing a typical small molecule, was capable of diffusing through the disuccinimidyl glutarate cross-linked hemoglobin membrane at 87.2% of its diffusion through buffer. PMID- 11495011 TI - Beneficial effects of Pluronic F-68 and artificial oxygen carriers on the post thaw recovery of cryopreserved plant cells. AB - The storage of prokaryotic and eukaryotic cells at ultra-low temperature in liquid nitrogen (-196 degrees C) is a procedure that has assumed an increasingly important role in underpinning many aspects of biotechnology. For eukaryotic cells, the transition from a cryopreserved state to physiologically normal temperatures and oxygen tensions, induces respiratory imbalances that may stimulate the production of toxic oxygen radicals causing impaired cellular functions. Novel treatments, that focus specifically on enhancing oxygen delivery to cells, are important in maximising post-thaw recovery. Recently, several approaches have been evaluated with suspension cultured plant cells as a model, yet biotechnologically-important, totipotent eukaryotic cell system. Such treatments include non-ionic surfactants, primarily Pluronic F-68, and artificial oxygen carriers, the latter based on inert perfluorochemical liquids or chemically-modifed haemoglobin, as supplements to culture medium used during the post-thaw recovery phase of cell growth. When used either alone or in combination, such novel treatments stimulate significantly the post-thaw viability and biomass production of cultured plant cells. Many of these technologies will be exploitable in cryopreservation protocols for eukaryotic cells in general. PMID- 11495012 TI - Pluronic F-68 enhanced shoot regeneration in a potentially novel citrus rootstock. AB - The effects have been studied in vitro of the non-ionic, co-polymer surfactant, Pluronic F-68, on shoot regeneration and bud induction in epicotyl and cotyledon explants of Citrus depressa, a potential alternative rootstock to C. jambhiri for commercial Citrus. Supplementation of Murashige and Skoog (1962)-based, agar solidified shoot regeneration/bud induction (SRBI) medium with 1.0 mg l(-1) 6 benzylaminopurine and 0.5% (w/v) Pluronic F-68 significantly (P < 0.05) increased mean fresh weight by a maximum of 60%, the proportion of explants exhibiting shoot/bud regeneration by 25% and the mean number of shoots per epicotyl explant by 184%, compared to untreated controls. Similarly, 0.5% (w/v) Pluronic F-68 significantly (P < 0.05) enhanced the mean percentage bud induction (91%) and the number of buds regenerated (>4-fold) per cotyledon explant. Interestingly, the mean fresh weight gain for both explants was unaffected across the range of concentrations (0.001-0.1% w/v) of Pluronic F-68 evaluated. Regenerated plants from epicotyl explants were transferred and acclimatized to glasshouse conditions. PMID- 11495013 TI - Bone tissue cultures: an in vitro model for the evaluation of bone defect healing after L-arginine and L-lysine administration. AB - The development of new therapeutic strategies and innovative biomaterials for the muscoloskeletal system, stresses the need for researchers to have reliable, easy, less time-consuming and ethical experimental models. The aim of the present study was to characterise an in vitro model of cultured rat femora and test the possibility of using this model in dynamic studies on bone healing. 24 femurs were explanted after 12 rats were killed for other experimental protocols. A standard bone defect was created in the distal femoral condyles and femurs were cultured in GBJb medium. Arginine and lysine were administered daily in the Arg Lys group. The other femurs were left untreated (Control group). At 1, 7, 14 and 21 days, alkaline phosphatase activity, nitric oxide and calcium were measured on the supernatant. At 21 days, femurs were embedded in polymethylmethacrylate for histomorphometry and microhardness evaluation of the newly formed bone. The current results showed that it was possible to study bone healing in vitro by using cultured bones from adult animals. A process for bone healing was observed also in untreated bones. Moreover, the structural analysis of the cultured bone showed that it had characteristics similar to those of the femurs, when they were embedded in resin immediately after animal sacrifice. The effect of Arg and Lys confirmed data of a previous study, where a faster healing of bone defect and fracture was observed in rabbits after Arg and Lys administration. PMID- 11495014 TI - Study of interaction of mannan-BSA neoglycoconjugates with Concanavalin A. AB - Neoglycoconjugates prepared by synthesis of oxidized mannans from Saccharomyces cerevisiae with bovine serum albumin were studied for interaction with Concanavalin A. The mannan-bovine serum albumin neoglycoproteins, different in degree of mannan oxidation used for synthesis and its content in conjugates were prepared. The interaction of these glycoconjugates with Concanavalin A by using precipitation method was investigated. The conjugates prepared at high weight ratio mannan: protein (4:1) involved 47-65% of saccharides and formed by precipitation with Concanavalin A aggregates with low content of protein. The obtained results showed that conjugates with lower content of mannan (up to 30%) are more efficacious for their aggregation with Concanavalin A than the conjugates with high content of mannan. PMID- 11495015 TI - Hemoglobin based oxygen carriers at a cross road: the old paradigms must be abandoned and much more basic science investigation is necessary. PMID- 11495016 TI - Sorption of herbicides in relation to soil variability and landscape position. AB - Using the soil-water sorption partitioning coefficient (Kd), this study quantified the spatial variation of 2,4-D sorption by soil in an undulating-to hummocky terrain landscape near Minnedosa, MB, Canada. Herbicide sorption was most strongly related to soil organic matter content and slope position, with greatest sorption occurring in lower landscape positions with greater soil organic matter content. The relation between sorption and slope position was more pronounced under conventional tillage (CT) than under long-term zero-tillage (ZT). Using multivariate regression and three independent variables (soil organic matter content, soil clay content and soil pH), the prediction of herbicide sorption by soil was very good for CT (R2 = 0.89) and adequately for ZT (R2 = 0.53). PMID- 11495017 TI - Sorption of atrazine and metolachlor by burrow linings developed in soils with different crop residues at the surface. AB - Atrazine and metolachlor sorption by earthworm (Lumbricus terrestris L.) burrows was measured by introducing herbicides into the burrows and collecting the effluent between 0 to 3, 3 to 6, and 6 to 9 min of simulated burrow flow. On average, sorption by burrow linings reduced the herbicide concentration to 78% (atrazine) and 74% (metolachlor) of the applied herbicide solution concentration. For both herbicides, the amount sorbed was dependent on the food source available to the earthworm, as well as the duration of burrow flow. On average, soybean-fed and corn-fed-earthworm-burrows significantly retained more herbicides relative to the Control Treatment (unfed-earthworms). More herbicides were transported through the burrows with time because the lateral flow movement from the burrow wall into the soil matrix decreased. It is also likely that herbicides retained on burrow linings during the first 3 min of flow saturated the adsorption sites on the burrow wall, which decreased the subsequent retention potential of herbicides in flow between 3 to 9 min. Based on these results, we conclude that herbicide transport through earthworm burrows in the field will be related to crop and crop residue management practices. PMID- 11495018 TI - Sorption behavior of metolachlor, isoproturon, and terbuthylazine in soils. AB - The sorption-desorption of metolachlor [2-chloro-N-(ethyl-6-methyl phenyl)-N-(2 methoxy-1-methyl ethyl) acetamide], isoproturon [3-(4-isopropyl phenyl)-1,1 dimethyl urea] and terbuthylazine [N6-tert butyl-6-chloro-N4-ethyl-1,3,5-triazine 2,4-diamine] herbicides was studied in two German soils at 1:10 soil to water ratio by batch method. Equilibrium of herbicides between soil and water (0.01 M CaCl2) was attained in 2 h. Sorption data fitted very well to Freundlich equation, represented by very high correlation coefficient (r2 > 0.934). Comparison of Freundlich K values indicated that sorption of all the three herbicides was most pronounced in soil having higher organic carbon content. Koc values were as expected nearly identical for each herbicide in the two soils. The Freundlich constant (1/n) was about 1 for metolachlor and less than 1 for terbuthylazine and isoproturon indicating a L-type of sorption isotherms. Desorption of all the three herbicides showed hysteresis. Nearly equal amounts of metolachlor, isoproturon and terbuthylazine were desorbed from both soils. There was a good correlation between Koc and solubility. PMID- 11495020 TI - Insecticide residues in cotton crop soil. AB - Dimethoate, monocrotophos, triazophos, deltamethrin, cypermethrin and endosulfan were applied to a cotton crop soil located at Nurpur village, Punjab, India. The insecticides were applied sequentially at recommended dosages in cotton fields by foliar application in 1995, 1996 and 1998. Soil samples were collected from the cotton crop farms and extracted with acetone. The extracted material was analysed by a gas liquid chromatograph (GLC) equipped with an 63Ni electron-capture detector (ECD-63Ni). Recovery data was obtained by fortifying soil with insecticide. The average recoveries from the fortified soil samples were 76-92% for organophosphorous compounds and 90-98% for synthetic pyrethroids and organochlorines. The results showed that the insecticide residues under study were present in the range of 1.16 to 41.97 ng g(-1) d.wt.soil. The pattern of dissipation of the insecticides used was similar for the duration of the crop. Half lives of the insecticides ranged from 7 to 22 days. Except endosulfan none of the other insecticides used were leached below 15 cm. Endosulfan was found to be rapidly degraded in the soil and formed a sulfate metabolite. Persistence and dissipation pattern in soils with history of exposure to the insecticide compared to non-history soils were similar. PMID- 11495019 TI - Dissipation of epoxiconazole in the paddy field under subtropical conditions of Taiwan. AB - The environmental fate and distribution of fungicide epoxiconazole were studied by a rice paddy field model ecosystem. One week before the head-sprouting stage, rice plant was treated separately once with OPUS (tradename of epoxiconazole) 12% SC 2.1 kg ha(-1) and 1.4 kg ha(-1), respectively. Soil, water and rice plant were sampled seven days intervals nine times after application. The bioconcentration factor of epoxiconazole on mosquito fish in the ecosystem was also determined, based on the amounts of epoxiconazole content both in fish and water. This was initiated one day after the fungicide treatment, and continued for four days. In addition, the residue of epoxiconazole in rice grains was analyzed after harvest. After harvest, both planted water spinach (Ipomoea aquatica Forsk) and edible amaranth (Amaranthus mangostanus L.) were analyzed. The results showed that epoxiconazole degraded in the local environment under the experimental conditions described. The degradation equations were in accordance with the first order kinetics. The DT50 of soil, field water and rice plant were 20-69 days, 11-20 days and 14-39 days, respectively. The bioconcentration factors of epoxiconazole on mosquito fish were 12.9 and 10.6 from 2.1 kg ha(-1) and 1.4 kg ha(-1) treatment, respectively. Residues of epoxiconazole in both rice and harvest vegetables were non-detectable. This indicates that epoxiconazole applied to rice at the recommended rates and application frequencies will not accumulate on rice grain and successive cropping vegetables. PMID- 11495021 TI - Sustainable soil management practices and quality of potato grown on erodible lands. AB - Land productivity can decline when top soil is lost. In Kentucky, limited resource farmers often produce vegetable crops on erodible lands. The objectives of this study were 1) to quantify the impact of three soil management practices (SMPs) on quantity of potato produced on erodible land, 2) to evaluate the impact of pyrethrin and azadirachtin insecticides on potato tuber quality, and 3) to assess the impact of yard waste compost on the chemical composition (ascorbic acid, free sugars, phenol contents) of potato tubers. Potatoes (Solanum tuberosum L. cv. Kennebec) were grown in a silty loam soil of 10% slope. Plots (n= 18) were 3.7 m wide and 22 m long (10% slope), with metal borders of 20 cm above ground level. Two botanical insecticides, Multi-Purpose Insecticide (containing pyrethrin 0.2%) and Neemix 4EC (containing 0.25% azadirachtin) were sprayed twice on potato foliage during each of two growing seasons (1997 and 1999) at the recommended rates of 6 lbs and 2 gallons.acre(-1), respectively. The SMPs were tall fescue strips (FS) intercropped between each two potato rows, soil mixed with yard waste compost (COM) and no-mulch (NM) treatment (roto-tilled bare soil). The experimental designwas a 2 x 3 x 3 factorial with main factors of two insecticides and three SMPs replicated three times. Average potato yield was lowest in NM and FS and highest in COM treatments. Yield obtained from the bottom of the plots was greater than that obtained from the top of plots. Tuber defects (rot, scab, sun green, hollow heart, necrosis, and vascular discoloration) were significantly different between the two growing seasons. The two insecticidal treatments did not have much influence on potato yield or tuber defects. Tubers obtained from tall fescue treatments had low levels of ascorbic acid and reducing sugars compared to compost treatments. PMID- 11495022 TI - Effects of phosphorothionate on the reproductive system of male rats. AB - Acute and Sub-acute toxic effects of a new pesticide phosphorothionate coded as RPR-V on testis of albino rat were studied. For the acute study, rats received a single dose of 30 mg/kg of RPR-V and sacrificed after 24 hours. For the Subacute study, 1.42 mg/kg/day was administered orally to rats for 10 days and 21 days. Acute exposure of rats to RPR-V brought no change either in the Gonadosomatic Index (GSI) or in the structure of testis or in the serum levels of Testosterone. Similarly, no significant change was observed in the Glutathione S-transferase (GST) activity. But, in testis there was significant increased in the reduced Glutathione (GSH) and Acid Phosphatase (AcP), whereas Alkaline Phosphatase (AkP) levels decreased significantly at 24hr post treatment. On 7th day (withdrawal period) after the cessation of the treatment the GSH, AcP, and AkP levels reached to near control. The sub-acute study revealed a significant post treatment. Due to RPR-V treatment the testis AcP levels increased significantly at 21st day of medication but AkP levels decreased both at 10th and 21st day of post treatment. Histopathological studies showed that after 10th day testis showed considerable loss of spermatozoids and at 21st day complete derangement of cellular organization was observed. Testosterone levels decreased significantly after 10th day and remained significantly low at 21st day. However, withdrawal studies showed a recovery in testis of rat treated with RPR-V. GST, GSH, GSI, AcP and AkP values were recovered, testosterone levels were also recovered but recovery in testis structure remained at a low profile. PMID- 11495023 TI - Population-level effects of the neem insecticide, Neemix, on Daphnia pulex. AB - Although natural insecticides from the neem tree are generally perceived as less harmful to the environment than synthetic insecticides, new evidence indicates that these products may pose a risk to certain nontarget organisms. In this paper, acute and chronic effects of commercial neem insecticides on the aquatic invertebrate, Daphnia pulex were examined. The acute toxicity of two commercial neem insecticides, Neemix, Azatin and the experimental insecticide, RH-9999 to D. pulex was investigated using traditional 48 hr concentration-mortality estimates. Neemix and Azatin were equitoxic with LC50's of 0.68 and 0.57 ppm; RH-9999 was significantly less toxic with an LC50 of 13 ppm. A 10 d population growth study was conducted for Neemix and a Neemix formulation blank (Neemix devoid of the active ingredients) to determine whether the active ingredients of Neemix and/or components of the formulation were responsible for toxicity. D. pulex populations went to extinction after exposure to a Neemix concentration of 0.45 ppm azadirachtin (equivalent to the acute LC7). Neemix No Observable Effect Concentration (NOEC) and Lowest Observable Effect Concentration (LOEC) values for population growth were 0.045 and 0.15 ppm azadirachtin, respectively. The mean number of offspring per surviving female (Ro) declined in a concentration dependent manner after exposure to Neemix with no offspring being produced after exposure to 0.45 ppm. Neemix NOEC and LOEC values for reproduction were 0.045 and 0.15 ppm, respectively. The formulation blank caused no mortality in the individuals used to start the population growth study but reduced reproduction and population growth accounting for 47% of the toxicity caused by Neemix at a concentration of 0.15 ppm. Thus, the formulation contributes substantially to the toxicity of Neemix but neem components are also toxic to D. pulex. Because the NOEC for population growth and reproduction were higher than the estimated environmental concentration of 0.035 ppm (a measure developed for forest pest mananagement), Neemix should pose little risk to populations of D. pulex. PMID- 11495024 TI - Spodoptera exigua: mating disruption, measurement of airborne concentration of pheromone, and use of specialist receptor cell responses for comparison to female pheromone emission. AB - Field experiments were conducted to evaluate Shin-Etsu Yoto-con-S 'rope' pheromone dispensers applied at different site densities in cotton to determine efficacy in disrupting sexual communication and mating of beet armyworm moths, Spodoptera exigua (Hubner); obtain atmospheric measurements of the disruptant pheromone components in and away from treated areas; and relate this information to estimates of the concentration of pheromone in the near vicinity of a calling virgin female moth. The number of pheromone-dispensing sites/ha ranged from 25 with 20 ropes/site up to a total of 247 sites with 2 ropes each. A total of 112 g pheromone of a 70:30 blend of (Z,E)-9,12-tetradecadien-l- ol acetate (ZETA) and (Z)-tetradecen-1-ol (Z9-14:OH) was applied/ha. As few as 25 sites/ha effectively disrupted sexual communication by 91%, as measured by reductions of moth captures in traps baited with pheromone lures, and mating by beet armyworm females by 94% for at least 10 weeks post-treatment. The results compared favorably to an earlier large-scale field experiment in which beet armyworm larval populations were reduced 95% using 500 pheromone-dispensing sites/ha (two ropes/site) and 160 g total pheromone/ha. The airborne concentration of ZETA and Z9-14:OH measured within the pheromone-treated plots that disrupted mating was 1.5 x 10(-13) M and 1.2 x 10(-13) M, respectively. The airborne concentration of ZETA and Z9-14:OH in the vicinity of a female was estimated (by excision and electrophysiological methods) to range between 2.1 x 10(-11) M and 2.4 x 10(-12) M, respectively, in a 1.6 km/h (1 mph) wind speed. Thus, the average ZETA concentration in the treated plots was about 140 times less than that expected from a single female; the average Z9-14:OH in the treated plots was about 20 times less. The data support an argument that disruption of mating by beet armyworm resulted from an imbalance in sensory input because mating was almost completely disrupted in all of the plots for at least 10 weeks with a mixture of ZETA:Z9-14:OH that differed significantly with that from excised female sex pheromone glands. Estimates of these parameters provide a benchmark for the requisite pheromone concentration in the atmosphere to achieve a high level of mating disruption for beet armyworm and possibly other insect pest species. PMID- 11495025 TI - Effects of selenium and mercury on the enzymatic activities and lipid peroxidation in brain, liver, and blood of rats. AB - Recent studies have reported on the toxicity and related oxidative stress of selenium and mercury. The present study compares the effects of Se as sodium selenite (Na2SeO3) and Hg as mercuric chloride (HgCl2) separately and in combination. Rats received repeated oral doses of Se (0.5 micromol/ml), Hg (0.5 micromol/ml), or Se in combination with Hg (0.5 micromol/ml of each) for 5 consecutive days. Rat serum, brain and liver samples were collected for biochemical assays. The following biochemical alterations occurred in response to Hg treatment: protein content (brain and liver), acetylcholinesterase (AChE) (brain and serum), acid and alkaline (AcP and AlP) phosphatases (plasma and liver) and glutathione S-transferase (GST) (plasma and liver) activities were significantly (P<0.05) decreased, while lactate dehydrogenase (LDH) (plasma, brain and liver), aspartate and alanine aminotransferase (AST, ALT) (serum and liver) activities were significantly increased. Thiobarbituric acid reactive substances (TBARS) was significantly increased in brain and liver. Effect of Se alone included decreased AcP, AlP and GST (serum and liver) activities. However, LDH (serum, brain and liver) and AST (liver) and TBARS (brain and liver) increased. Selenium in combination with Hg partially or totally alleviated the toxic effects of Hg on different studied enzymes. It is concluded that Se could be able to antagonize the toxic effects of mercury. PMID- 11495026 TI - Absence of apolipoprotein E4 genotype, good gallbladder motility and presence of solitary stones delay rather than prevent gallstone recurrence after extracorporeal shock wave lithotripsy. AB - BACKGROUND/AIMS: Extracorporeal shock wave lithotripsy (ESWL) with adjuvant bile salt dissolution therapy may be successful in selected gallstone patients, but the considerable risk of recurrence is a major drawback. Apolipoprotein E4 genotype and impaired gallbladder motility have been identified as major risk factors for recurrence during short-term follow up. We have now examined their relevance during long-term follow up. METHODS: Eighty-four cholesterol gallstone patients (55 solitary and 29 multiple (two to ten) stones) were followed prospectively up to 10 years after complete stone disappearance. Various potential risk factors for recurrence were evaluated. RESULTS: Gallstone recurrence was found in up to 80% of patients at 10 years follow-up. Absence of the apolipoprotein epsilon4 allele, initial solitary stones, good gallbladder emptying (i.e. minimal postprandial volume < or = 6 ml) and 2-year postdissolution ursodeoxycholic acid prophylaxis (in ten patients) all delayed but did not prevent recurrence. In contrast, regular use of non-steroidal anti inflammatory drugs (NSAIDs) was identified as an independent protective factor, with greatly decreased recurrence (at 10 years: 58 vs 93% in non-NSAID users, P = 0.03). CONCLUSIONS: Non-apolipoprotein E4 genotype, presence of solitary stones and good gallbladder emptying delay rather than prevent recurrence after initially successful ESWL. Regular use of NSAIDs may prevent recurrence. PMID- 11495027 TI - T-helper cell response to woodchuck hepatitis virus antigens after therapeutic vaccination of chronically-infected animals treated with lamivudine. AB - BACKGROUND/AIMS: Immunotherapy of patients chronically-infected with hepatitis B virus (HBV) may have the risk of fulminant hepatitis. This risk might be diminished if immunotherapy was carried out under conditions of low viremia. METHODS: Five woodchucks chronically-infected with woodchuck hepatitis virus (WHV), a virus closely related to HBV, were treated with lamivudine for 23 weeks. At week 10, when viremia had decreased by 3-5 logs, three woodchucks were vaccinated with woodchuck hepatitis virus surface antigen (WHsAg) plus the T helper determinant FISEAIIHVLHSR. RESULTS: It was found that the administration of lamivudine only, had no effect on the T-helper response against WHV antigens. By contrast, vaccination induced T-helper responses against WHV antigens, shifting the cytokine profile from Th2 to Th0/Th1, but was without effect on viremia, WHsAg levels, or anti-WHs antibodies. Analysis of liver biopsies showed that lamivudine administration may have reduced hepatic inflammation. By contrast, vaccination clearly enhanced hepatic inflammation. After lamivudine withdrawal, viremia returned to high levels. CONCLUSIONS: These results suggest that therapeutic vaccination of chronically-infected woodchucks under conditions of low viremia shifts the cytokine profile against viral antigens towards Th0/Th1. This shift may prevent the efficient induction of anti-WHs antibodies. PMID- 11495028 TI - Analysis of full-length hepatitis A virus genome in sera from patients with fulminant and self-limited acute type A hepatitis. AB - BACKGROUND/AIMS: Type A hepatitis still poses a considerable problem worldwide. Why some patients progress to fulminant type A hepatitis and others do not is still unknown. To examine whether genomic differences of hepatitis A virus (HAV) are responsible for the severity of the disease, we analyzed the whole HAV genomes from patients with fulminant and self-limited acute type A hepatitis. METHODS: Sera from three patients with sporadic type A fulminant hepatitis (FH) and three patients with acute hepatitis (AH) were examined for HAV RNA. Full length nucleotide sequences were determined using long reverse transcription polymerase chain reaction, 5' and 3' rapid amplification of cDNA ends methods, and direct sequencing. The amino acid sequences were deduced from the nucleotide sequences. RESULTS: HAV RNA was detected in all six patients examined. From the sequence of viral protein 1/2A, all cases were revealed to be genotype IA. By comparing with genotype IA, wild-type HAV strain GBM, the analysis of whole genomes from the six cases showed no specific substitutions between FH and AH. Completely identical nucleotide sequences were observed at 3' non-translated region (NTR) in all six cases. In 5'NTR, less nucleotide substitutions were found in FH than in AH, and in the non-structural protein 2B region, a little more amino acid substitutions seemed to be found in FH than in AH. CONCLUSIONS: This study showed that full-length HAV could be analyzed from serum samples. Although there were no unique nucleotide or amino acid substitutions, possible associations were suggested between the severity of type A hepatitis and the nucleotide substitutions in 5'NTR and the amino acid substitutions in 2B. PMID- 11495029 TI - Changes in histological lesions and serum fibrogenesis markers in chronic hepatitis C patients non-responders to interferon alpha. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the changes in histological lesions and serum N-terminal peptide of type III procollagen (PIIINP) and hyaluronate (HA) levels in virologic non-responder patients treated by Interferon alpha (IFNalpha). METHODS: We enrolled 183 patients treated by IFNalpha and 56 controls, all with paired biopsy specimens. Yearly liver fibrosis progression was estimated before and during a follow-up of 1 year. RESULTS: By contrast to sustained responders, non-responders (n = 105) did not achieve improvement of histological scores after therapy. Their yearly fibrosis progression rate was similar before and during follow-up (0.18, 95%CI: 0.16-0.20, vs 0.26 (95%CI: 0.12-0.40) fibrosis units/year, NS), and was not different in controls (0.17, 95%CI: 0.06-0.27). The levels of PIIINP and hyaluronate (HA) remained unchanged during follow-up. Histological improvement was observed for the second biopsy in 25% of non-responders, but also in 23% of controls. This improvement was not correlated with decrease of ALT level, viral load, PIIINP, or HA. CONCLUSIONS: Our results suggest that IFNalpha therapy is unable to decrease PIIINP or HA levels and cannot improve the histological outcome in virologic non responder patients. The histological improvement observed in a subset of patients may be linked to sample fluctuation or lack of reproducibility of histological scores. PMID- 11495030 TI - Unravelling complexity--the scientific basis of genetic predisposition in autoimmune liver disease. PMID- 11495031 TI - Alcoholic liver disease: a matter of hormones? PMID- 11495032 TI - Ursodeoxycholic acid 'mechanisms of action and clinical use in hepatobiliary disorders'. AB - UDCA exerts its beneficial effect in liver diseases through a diverse, probably, complementary array of mechanisms. The clinical use and efficacy of UDCA in PBC have been evident. UDCA may also have a place in the management of PSC, ICP, cystic fibrosis, PFIC and GVHD involving the liver, although, more studies are needed to further determine its therapeutic potential in these diseases and in other hepatobiliary disorders such as liver allograft rejection, drug and TPN induced cholestasis, NASH, and alcoholic liver disease. PMID- 11495033 TI - Images in hepatology. Acute schistosoma mansoni infection in the liver. PMID- 11495034 TI - Heterozygosity for a polymorphism in the promoter region of the UGT1A1 gene. PMID- 11495035 TI - The intrahepatic T helper cytokine profile does not predict the response to alpha interferon in chronic hepatitis C. PMID- 11495036 TI - Diabetic KKAy mice exhibit increased hepatic PPARgamma1 gene expression and develop hepatic steatosis upon chronic treatment with antidiabetic thiazolidinediones. AB - BACKGROUND/AIMS: Peroxisome proliferator-activated receptor-gamma, which is involved in the regulation of lipid homeostasis, is upregulated in the liver of obese and diabetic mice, but the biological consequences of this induction are largely unknown. This study was aimed at further characterizing this upregulation and exploring the downstream biological effects of specific activators on hepatic lipid metabolism. METHODS: Hepatic expression of peroxisome proliferator activated receptor-gamma1 and gamma2 mRNA and protein was analyzed by real-time polymerase chain reaction and Western immunoblotting in KKAy mice and ob/ob mice. KKAy mice were treated with thiazolidinediones, and hepatic triglyceride content and lipid distribution were analyzed biochemically and by histopathology. RESULTS: KKAy mice exhibited a marked increase in hepatic peroxisome proliferator activated receptor-gamma1 mRNA and protein levels, whereas the gamma2 isoform was upregulated in ob/ob mice. Treatment of KKAy mice with troglitazone or rosiglitazone resulted in severe microvesicular periacinar steatosis, whereas lean control mice did not develop any pathological liver changes. Hepatic triglyceride levels, however, were not altered by the treatment. CONCLUSIONS: In mice with obesity-associated upregulated hepatic peroxisome proliferator activated receptor-gamma expression, thiazolidinediones may produce hepatic steatosis. Under pathophysiological conditions, such as non-insulin-dependent diabetes, the liver may thus become sensitized towards peroxisome proliferator activated receptor-gamma-activating drugs. PMID- 11495037 TI - Ductular morphogenesis and functional polarization of normal human biliary epithelial cells in three-dimensional culture. AB - BACKGROUND/AIMS: The understanding of the physiology and function of human biliary epithelial cells (hBEC) has been improved by studies in monolayer culture systems. The aim was to develop a polarized model to elucidate the mechanisms of ductular morphogenesis and functional differentiation of hBEC. METHODS: The morphological, phenotypic and functional properties of hBEC cultured as three dimensional aggregates in collagen gel were assessed in medium supplemented with (or without) human hepatocyte growth factor (hHGF) and foetal bovine serum. RESULTS: In the absence of added mitogens and serum, cells maintained as morphologically polarized aggregates, organized around a central lumen, were positive for phenotypic markers of biliary epithelium and negative for markers of other cell types. Functional markers, gamma-glutamyl-transferase, anion exchanger 2, responses to gamma interferon and forskolin induced secretion, were preserved. hHGF increased both the size and number of aggregates and induced hBEC to invade the gel and lumena forming anastomosing networks of cells. CONCLUSIONS: Collagen gel culture in the absence of added growth factors and serum provides a model for analysis of the polarized functions of hBEC. The formation of poorly organized cords of cells in response to hHGF suggests that collagen gel culture may provide a model for the investigation of atypical ductular morphogenesis of the human biliary tract. PMID- 11495038 TI - Autoimmune hepatitis in Brazilian patients is not linked to tumor necrosis factor alpha polymorphisms at position -308. AB - BACKGROUND/AIMS: Susceptibility to autoimmune hepatitis (AIH) has been linked to different HLA-DR antigens. Recently, AIH type 1 was associated with polymorphisms in the tumor necrosis factor alpha gene promoter (TNFA) at position -308. In this respect, the frequency of the TNFA*2 allele, in linkage disequilibrium with HLA DRB1*0301, was shown to be significantly increased in whites with AIH type 1. The aim of this study was to assess the role of TNFA alleles in conferring susceptibility to AIH, studying a population where the disease is not primarily associated with HLA-DRB1*03. METHODS: The determination of HLA-DRB1 and TNFA alleles was performed in 92 patients with AIH type 1, 29 subjects with AIH type 2 and 83 healthy controls by polymerase chain reaction-based techniques. RESULTS: The distribution of TNFA alleles was similar in patients with AIH types 1 and 2, when compared with controls. In addition, the TNFA*2 allele was identified in patients carrying HLA-DR antigens other than HLA-DRB1*03. Interestingly, higher gammaglobulin levels were observed in TNFA*2 positive patients. CONCLUSIONS: Our data indicate that susceptibility to AIH remains primarily linked to the HLA-DRB1 locus, and suggest that the association of AIH with TNFA*2 previously observed in whites might be secondary to a linkage disequilibrium with HLA-DRB1*0301. PMID- 11495040 TI - Prevalence and prognostic value of quantified electroencephalogram (EEG) alterations in cirrhotic patients. AB - BACKGROUND/AIMS: The electroencephalogram (EEG) is frequently altered in cirrhotic patients. We, therefore, performed a study to ascertain the features and the prognosis of cirrhotic patients without current overt hepatic encephalopathy (OHE) who have EEG alterations. METHODS: A series of 296 consecutive cirrhotic patients who had undergone quantified-EEG was studied. The median follow-up was 442 days, 128 patients had bouts of OHE and 78 patients died from liver-related causes. Another group of 124 cirrhotic patients with a median follow-up of 223 days was examined to validate the prognostic model. RESULTS: EEG alterations were detected in 38% of the patients. The prevalence of EEG alterations was associated with the severity of cirrhosis (class B: odds ratio (OR) = 2.3, 95% confidence interval (CI) = 1.2-4.7; class C: OR = 3.5, 95% CI = 1.6-7.7), but not with the aetiology (alcoholic vs. non-alcoholic: OR = 0.9; 95% CI = 0.5-1.5). The EEG predicted the occurrence of OHE (chi2 = 26; P < 0.001) and mortality (chi2 = 34; P < 0.001), also adjusting for Child-Pugh class by a multivariate analysis. In the patients with a Child-Pugh score of > or = 8, the EEG discriminated between those patients with a higher 1-year risk of OHE (hazard ratio (HR) = 3.3, 95% CI = 1.8-6.1) and death (HR = 3.1, 95% CI = 1.7-5.6). CONCLUSIONS: In conclusion, quantified-EEG had a prognostic value for the occurrence of bouts of OHE and mortality in cirrhotic patients. PMID- 11495039 TI - Altered endothelin receptor expression in prehepatic portal hypertension predisposes the liver to microcirculatory dysfunction in rats. AB - BACKGROUND/AIMS: Endothelin (ET) is one of the most active vascular regulators in the liver. It is unknown how partial portal vein ligation (PPVL) induced prehepatic portal hypertension influences the response of the liver to ET and its agonists. Therefore, this study was conducted to determine the expression of ET receptors and its functional significance after PPVL. METHODS: Competitive receptor binding study and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) were performed using liver homogenates after 2 weeks of PPVL or sham operation in rats. Hepatic microcirculation was evaluated in vivo using intravital microscopy. RESULTS: Although there was no significant difference in dissociation constant (Kd) and total amount of receptors (Bmax) between sham and PPVL, the proportion of ET(B) receptor was significantly increased in PPVL. RT-PCR analysis confirmed the up-regulation of ET(B) receptors demonstrated by the competitive receptor binding assay. In the functional study, infusion of ET(B) agonist (IRL 1620) in a low dosage did not change the hepatic microcirculation in sham but strongly constricted the sinusoids leading to a reduction of sinusoidal perfusion in PPVL. CONCLUSIONS: These results suggest that prehepatic portal hypertension may predispose the hepatic microcirculation to dysregulation in stress conditions where ET is upregulated. PMID- 11495041 TI - The antiestrogen toremifene protects against alcoholic liver injury in female rats. AB - BACKGROUND/AIMS: Females are generally considered to be more susceptible to alcohol-induced liver injury than males. To elucidate whether gonadal hormones are involved, female rats were chronically treated with ethanol and with an antiestrogen. METHODS: Ethanol was administered in a low-carbohydrate liquid diet. Estrogen action was blocked by daily intubation of toremifene, a non hepatotoxic second generation estrogen receptor antagonist. RESULTS: The female rats consuming intoxicating amounts of ethanol diet for 6 weeks developed massive microvesicular/macrovesicular steatosis, frequent inflammatory foci and spotty necrosis. Serum alanine aminotransferase increased 7-fold. Toremifene treatment did not affect steatosis, but significantly reduced inflammation and necrosis. Ethanol increased the expression of CD14 and tumor necrosis factor- (TNF) alpha mRNA and also the production of TNF-alpha by isolated Kupffer cells, but toremifene had no significant counteracting effect. However, toremifene significantly alleviated both ethanol induction of the pro-oxidant enzyme CYP2E1 and ethanol reduction of the oxidant-protective enzyme Se-glutathione peroxidase. CONCLUSIONS: The partial protection by toremifene against ethanol-induced liver lesions suggests a pathogenic contribution of estrogens, possibly associated with an oxygen radical mediated mechanism. PMID- 11495042 TI - A non-toxic heat shock protein 70 inducer, geranylgeranylacetone, suppresses apoptosis of cultured rat hepatocytes caused by hydrogen peroxide and ethanol. AB - BACKGROUND/AIMS: A stress-inducible heat shock protein 70 (HSP70) is one of the best-known endogenous factors protecting cell injury under various pathological conditions. The aim of this study was to examine anti-apoptotic actions of a non toxic HSP70 inducer, geranylgeranylacetone (GGA), on hepatocytes exposed to hydrogen peroxide (H2O2) or ethanol. METHODS: Primary cultures of rat hepatocytes were treated with different concentrations of GGA and exposed to 0.5 mM H202 or 100 mM ethanol. The heat shock response was assessed by measuring the activation of heat shock factor 1 (HSF1), HSP70 mRNA expression, and accumulations of HSP70, HSP90, and HSP27. Apoptosis was evaluated by DNA fragmentation. RESULTS: Pretreatment with 1 microM GGA for 2 h enhanced nuclear translocation and phosphorylation of HSF1, HSF1-DNA binding, HSP70 mRNA expression, and its accumulation, when the cells were exposed to H202 or ethanol. In association with this accelerated response, GGA suppressed the insult-induced activation of c-Jun N-terminal kinases, caspase 9, and caspase 3-like proteases, leading to significant inhibition of apoptosis. CONCLUSIONS: GGA exerted anti-apoptotic actions, at least in part, by priming hepatocytes for enhanced HSP70 induction. Our results suggest that GGA may have a potential benefit for the treatment of alcoholic and ischemia-reperfusion liver injuries. PMID- 11495043 TI - p16INK4A-alterations in primary angiosarcoma of the liver. AB - BACKGROUND/AIMS: Alterations in the p16 (CDKN2/MTS-1/INK4A) gene have been implicated in the tumorigenesis of different human cancers. Recent evidence shows that transcriptional silencing as a consequence of hypermethylation of CpG islands is the predominant mechanism of p16INK4a gene inactivation in malignant epithelial tumors. This study was performed to determine whether alterations of p16 are involved in the development of angiosarcoma of the liver. METHODS: The status of p16 was evaluated in 17 angiosarcomas of the liver by methylation specific PCR (MSP), microsatellite analysis, DNA sequencing and immunohistochemical staining. The results obtained were correlated with histopathological variables and with patient survival. RESULTS: Hypermethylation of the 5' CpG island of the p16 gene was found in 12 out of 17 (71%) angiosarcomas examined. Homozygous deletion at the p16 region was present in one case (6%), and loss of heterozygosity was present in two cases (12%). We failed to detect p16 gene missense mutations. The status of p16 correlated with neither histopathological factors nor with the prognosis of the patients with angiosarcomas. CONCLUSIONS: These data suggest that inactivation of the p16 gene is a frequent event in angiosarcomas of the liver. The most common somatic alteration is promotor methylation of the p16 gene. We failed to establish p16 as independent prognostic factors in these tumors. PMID- 11495044 TI - Activation of c-Src gene product in hepatocellular carcinoma is highly correlated with the indices of early stage phenotype. AB - BACKGROUND/AIMS: The aim of this study was to investigate whether c-Src is involved in carcinogenesis and progression of human hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma. METHODS: We designed an immunohistochemical study using Clone 28, an antibody that specifically recognizes the activated form of c-Src. RESULTS: Hepatocytes in normal liver, chronic hepatitis with or without cirrhosis, atypical adenomatous hyperplasia, as well as bile ductular cells, and infiltrating mononuclear cells were all negative for immunohistochemical staining for the activated c-Src. Among 87 cases of HCC tested, 40 (46%) were positively stained for the activated c-Src, and this positive staining was inversely correlated with the Ki-67 labeling index (LI) (P = 0.0031), intrahepatic metastasis (P = 0.0099), TNM stage (P = 0.0062), alpha fetoprotein (P = 0.0103) and epidermal growth factor-receptor expression (P = 0.0153). Positive staining for the activated c-Src was more frequently observed in well- or moderately-differentiated carcinoma (P = 0.0256). In multivariate analysis, the activated c-Src expression was independently related to the Ki-67 LI (P = 0.0197). In contrast to positive staining in HCC, cholangiocarcinoma were classified as negative in all 19 cases examined. CONCLUSIONS: These results strongly suggest the involvement of activated c-Src in early stages of HCC, and suggest that cholangiocarcinoma might employ different signaling mechanisms. PMID- 11495045 TI - The role of overexpression and gene amplification of cyclin D1 in intrahepatic cholangiocarcinoma. AB - BACKGROUND/AIMS: Intrahepatic cholangiocarcinoma (ICC) is a primary liver malignant tumor with an extremely poor prognosis, but less attention has been directed to factors related to molecular carcinogenesis, including cell cycle proteins. We examined the expression and gene amplification of cyclin D1, the cell cycle regulating protein. Our objective was to evaluate correlations with clinicopathological factors in ICC. METHODS: Cyclin D1 overexpression and cellular proliferative activity (Ki-67 labeling index) were investigated immunohistochemically, and 20 cases were further investigated for cyclin D1 gene amplification, using differential PCR. We examined the correlation between the expression and gene amplification of cyclin D1 and clinicopathological factors, including overall survival in patients with ICC. RESULTS: Immunohistochemical analysis revealed an overexpression of cyclin D1 protein in 28 of 66 subjects with ICCs (42%). The cyclin D1 overexpression was associated with poor histological differentiation (P = 0.04), high cellular proliferative activity (P < 0.01), and a poor prognosis (P = 0.02) by univariate analysis, although it is not an independent prognostic factor by multivariate analysis. Cyclin D1 gene amplification was confirmed in five of the 20 patients. Of those five cases of ICC, all had poor histological differentiation, and four of the five ICCs (80%) showed evidence of cyclin D1 immunoreactivity. CONCLUSIONS: Overexpression and gene amplification of cyclin D1 are frequent and contribute to dedifferentiation and cellular proliferative activity of ICCs, and overexpression also indicates a poor prognosis for patients with ICC. PMID- 11495046 TI - Pancreastatin, a chromogranin A-derived peptide, inhibits DNA and protein synthesis by producing nitric oxide in HTC rat hepatoma cells. AB - BACKGROUND/AIMS: Pancreastatin, a chromogranin A-derived peptide, has a counter regulatory effect on insulin action. We have previously characterized pancreastatin receptor and signalling in rat liver and HTC hepatoma cells. A G alpha(q/11)-PLC-beta pathway leads to an increase in [Ca2+]i, PKC and mitogen activated protein kinase (MAPK) activation. These data suggested that pancreastatin might have a role in growth and proliferation, similar to other calcium-mobilizing hormones. METHODS: DNA and protein synthesis were measured as [3H]-thymidine and [3H]-leucine incorporation. Nitric oxide (NO) was determined by the Griess method and cGMP production was quantified by enzyme-linked immunoassay. RESULTS: Contrary to the expected results, we have found that pancreastatin inhibits protein and DNA synthesis in HTC hepatoma cells. On the other hand, when the activity of NO synthase was inhibited by N-monomethyl-L arginine (NMLA), the inhibitory effect of pancreastatin on DNA and protein synthesis was not only reverted, but a dose-dependent stimulatory effect was observed, probably due to MAPK activation, since it was prevented by PD98059. These data strongly suggested the role of NO in the inhibitory effect of pancreastatin on protein and DNA synthesis, which is overcoming the effect on MAPK activation. Moreover, pancreastatin dose-dependently increased NO production in parallel to cyclic guanosine monophosphate (cGMP). Both effects were prevented by NMLA. Finally, an indirect effect of pancreastatin through the induction of apoptosis was ruled out. CONCLUSIONS: Therefore, the NO and the cGMP produced by the NO-activated guanylate cyclase may mediate the dose-dependent inhibitory effect of pancreastatin on growth and proliferation in HTC hepatoma cells. PMID- 11495047 TI - Short-term effects of statin therapy in patients with hyperlipoproteinemia after liver transplantation: results of a randomized cross-over trial. AB - BACKGROUND/AIMS: Hyperlipoproteinemia is frequent following liver transplantation and may lead to atherosclerosis. Lipid-lowering agents may be useful, but could interfere with the function of the transplanted organ and with immunosuppression. We therefore evaluated in a prospective, randomized, open-labeled cross-over trial the effect of two frequently used 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (pravastatin 10 mg d(-1) and cerivastatin 0.1 mg d(-1)) in hyperlipoproteinemic patients after liver transplantation. METHODS: Sixteen patients (6.3 +/- 2.0 years post-transplantation, cyclosporine n = 11, tacrolimus n = 5) with hyperlipoproteinemia (cholesterol 246 +/- 42, triglycerides 191 +/- 87, low-density lipoprotein (LDL)-cholesterol 161 +/- 35, high-density lipoprotein (HDL)-cholesterol 44 +/- 11 mg d(-1)) were included. Treatment periods of 6 weeks were separated by a 4-week washout period. RESULTS: Both medications were tolerated well, no effects on serum concentrations of liver enzymes or immunosuppressive agents were observed. Cerivastatin and pravastatin decreased (P < 0.001) cholesterol by 21 +/- 10% and 15 +/- 10%, LDL-cholesterol by 27 +/- 14% and 17 +/- 15%, respectively, while triglyceride and HDL cholesterol concentrations did not change significantly. LDL/HDL-cholesterol markedly improved (P < 0.001) by 29 +/- 16% (cerivastatin) and 16 +/- 16% (pravastatin). Cerivastatin was more potent than pravastatin in patients receiving cyclosporine A, while there was no significant difference in patients receiving tacrolimus. CONCLUSIONS: Low-dose cerivastatin and pravastatin significantly improve lipid profiles following liver transplantation without affecting liver function or immunosuppression. PMID- 11495048 TI - Emergence of YMDD motif mutant of hepatitis B virus during short-term lamivudine therapy in South Korea. AB - BACKGROUND/AIMS: The emergence of a YMDD mutant resistant to lamivudine therapy has been reported in patients with hepatitis B treated with long-term lamivudine therapy. However, it is not well known whether the YMDD mutant could be detected early in lamivudine therapy in hepatitis B virus (HBV) endemic areas. The aim of this study was to investigate the emergence of the YMDD mutant during short-term lamivudine therapy in South Korea. METHODS: We prospectively investigated the emergence of the YMDD mutant by the nested PCR assay using restriction fragment length polymorphism in 28 patients with chronic hepatitis B who were treated with 100 mg of lamivudine daily for 12 weeks. RESULTS: The YMDD mutant was detected in 17 (60.7%) out of 28 patients at week 12, and the only type of mutation found was the YIDD mutation. When we carried out the nested PCR serially in five patients, YIDD mutants were detected as early as 2 weeks by the nested PCR assay. The nested PCR results were in concordance with DNA sequencing in one patient's serial samples. CONCLUSIONS: YMDD mutants in HBV were detected within a few weeks during lamivudine therapy in South Korea, which suggests that the YMDD mutant may exist even before lamivudine therapy in HBV endemic areas. PMID- 11495050 TI - Modification and characterization of artificially patterned enzymatically active surfaces by scanning electrochemical microscopy. AB - This review summarizes the characterization of localized enzymatic activity by scanning electrochemical microscopy (SECM). After introducing the concepts of feedback imaging and generator-collector experiments with enzyme-modified solid surfaces, a comparison of the merits and limitations of both approaches is given and further illustrated by selected applications. They include enzyme-modified patterned monolayers, enzyme-modified polymer microstructures and enzyme-modified metal microstructures. Such configurations are important for the development of miniaturized bioanalytical systems with proteins, such as miniaturized enzyme electrode arrays. SECM has emerged as an ideal tool for prototyping of such systems. It also offers several mechanisms for local surface modifications under conditions compatible with conservation of protein functionality of enzymes and antibodies. The subsequent imaging of the immobilized activity provides direct information about local immobilized enzyme activity. The range of biotechnological applications can be expanded by labeling other biomolecules, such as monoclonal antibodies, with appropriate enzymes. Miniaturized electrochemical enzyme immunoassays that apply the sandwich format and SECM as the detection method are reviewed. They have been performed on microstructured supports after reagent spotting or on agglomerates of surface-modified magnetic microbeads. Finally, current challenges are listed with indications of ongoing research to overcome current limitations by means of instrumental improvements. PMID- 11495049 TI - Transmission of hepatitis B virus infection in Gambian families revealed by phylogenetic analysis. AB - BACKGROUND/AIMS: Transmission of hepatitis B virus (HBV) in Africa occurs horizontally, with most people becoming infected between the ages of 1 and 5 years. The index cases in such events have been assumed to come from within the family unit or from sources outside the immediate family, such as other families or inhabitants of the same compound or village. Here, we define these routes of transmission by phylogenetic tree analysis of sequences from the entire pre core/core region of the virus, in Gambian chronic carriers. METHODS: Amplification by polymerase chain reaction of serum extracted HBV-DNA was followed by direct sequencing of the target region. Following editing and alignment of these sequences, phylogenetic tree analysis was performed using the neighbour-joining and maximum-likelihood methods. RESULTS: Despite the overall conserved nature of the sequences of the pre-core/core region from 142 chronic carriers, distinct clusters were easily defined at the family and village level, but not on a wider geographical separation. CONCLUSIONS: Phylogenetic tree analysis of sequences obtained from family members provided strong evidence of intrafamilial transmission of HBV in at least two-thirds of the families studied from Gambia. PMID- 11495051 TI - Separation of metal ions by capillary electrophoresis--diversity, advantages, and drawbacks of detection methods. AB - Capillary electrophoresis (CE) has been applied to metal-ion analysis during the last 10 years. To improve sensitivity and selectivity different modes of detection have been adapted or developed. The selection of commercially available detection systems for metal-ion analysis is still primarily limited to UV-Vis detection, although other commercial systems, e.g. fluorescence, conductivity, or interfaces for coupling to mass spectrometry (MS) or inductively coupled plasma mass spectrometry (ICP-MS) are becoming available. High demands are made on any detector used in CE, because the analytical signal has to be extracted from less than 1 nL of sample, which corresponds to a total amount of < or = 10(-12) to 10( 15) mol analyte. This paper compares currently available and recently developed detection methods for CE as applied to the analysis of metal ions. Commercially available techniques, for example UV-Vis, fluorescence, or mass spectrometry, and other new detection methods including electrochemistry, radioactivity, and XRF, are discussed and future trends are anticipated. PMID- 11495052 TI - Laser ablation--reflections on a very complex technique for solid sampling. AB - This paper is an attempt to point out the complex correlations between the experimental conditions in solid sampling by lasers. In particular, the influence of the laser properties, the surrounding gas, and the matrix on the analytical results of laser ablation techniques, such as laser induced breakdown spectrometry or laser ablation-ICP-MS, will be discussed. PMID- 11495053 TI - Self-absorption effects in radially and axially viewed inductively coupled plasma atomic emission spectrometry--the key role of the operating conditions. AB - Self-absorption effects leading to curvatures of the upper part of calibration graphs were investigated in multichannel detection ICP-AES. A dual view Optima 3000 ICP system was used to enable the simultaneous determination of 38 lines for both radial and axial viewing. Resonance and non-resonance lines were selected for both atomic and ionic lines. The concentrations of 22 standards were in the range 0.1-100 mg L(-1) and two sets of operating conditions, namely power and carrier gas flow rate, were used to evaluate their influence. It was found that these two conditions, and in particular the carrier gas flow rate, play a major role in self-absorption effects. Except for strongly absorbing lines, it was possible, under suitable conditions, to reduce or to suppress differences between self-absorption effects in radial and axial viewing, enabling extension of the range of linearity of axial viewing to higher concentrations. A diagnostic tool, based on emission line ratios, is proposed for detection of self-absorption. A calibration procedure is given for strongly absorbing lines affected by self absorption even when operating conditions were optimized. PMID- 11495054 TI - Role of electronegativity in the qualitative inference of the TOF-SIMS fragment pattern of inorganic compounds. AB - The role of the electronegativity of atoms in inorganic compounds in TOF-SIMS fragmentation is discussed. From a study of approximately 30 inorganic compounds- chlorides, oxides, nitrates, and sulfates--a simple rule has been proposed for the dependence of fragment pattern appearance on the electronegativity (electron affinity), which can be easily obtained from handbooks, and the valence of positive and negative ions in these compounds. TOF-SIMS measurements of metal and alloy surfaces, should be corrected for the ionization potentials and/or electronegativities of atoms present in surface contaminants. PMID- 11495055 TI - An interlaboratory study to test instrument performance of hydrogen dual-inlet isotope-ratio mass spectrometers. AB - An interlaboratory comparison of forty isotope-ratio mass spectrometers of different ages from several vendors has been performed to test 2H/1H performance with hydrogen gases of three different isotopic compositions. The isotope-ratio results (unsufficiently corrected for H3+ contribution to the m/z = 3 collector, uncorrected for valve leakage in the change-over valves, etc.) expressed relative to one of these three gases covered a wide range of values: -630% to -790% for the second gas and -368% to -462% for the third gas. After normalizing the isotopic abundances of these test gases (linearly adjusting the delta values so that the gases with the lowest and highest 2H content were identical for all laboratories), the standard deviation of the 40 measurements of the intermediate gas was a remarkably low 0.85%. It is concluded that the use of scaling factors is mandatory for providing accurate internationally comparable isotope-abundance values. Linear scaling for the isotope-ratio scales of gaseous hydrogen mass spectrometers is completely adequate. PMID- 11495056 TI - New cyclodextrin derivatives as chiral selectors in capillary electrophoresis. AB - The separation of three pairs of enantiomeric herbicides has been successfully achieved by capillary electrophoresis at two different pH values in the presence of cyclodextrin derivatives previously synthesized in our laboratory. Two of these derivatives constitute a new class of receptor, the hemispherodextrins, in which a trehalose capping moiety is bonded to beta-cyclodextrin. Because of their peculiar structure hemispherodextrins have very promising characteristics and the low receptor concentration required to achieve separation deserves particular interest. PMID- 11495057 TI - Experimental verification of the properties of ion-selective electrodes for low concentration determination. AB - The influence of several ion-selective electrode properties on electrode response and selectivity at low concentration levels has been investigated experimentally. The properties investigated were the composition of inner electrode solution, the composition of the membrane, the presence of interfering ions in the sample, and the thickness of diffusion layer in the sample solution. All the results obtained confirmed theoretical considerations. PMID- 11495058 TI - Thin-layer chromatographic behavior of rare earths on silica gel with aqueous alkaline earth metal nitrate solutions as mobile phases. AB - The TLC behavior of all the rare earths except Pm has been surveyed on silica gel (pH 7.0) pretreated with 0.1 mol L(-1) tris(hydroxymethyl)aminomethane and 0.1 mol L(-1) HCl with aqueous nitrate solutions of alkaline earth metals as mobile phases. The RF values of the lanthanides varied in a regular and characteristic way accompanied by the tetrad effect with increasing atomic number, and when the mobile phases were changed the RF values of each metal decreased in the order Mg2+ > Ca2+ > Sr2+ > Ba2+, as the crystal ionic radii of the alkaline earth metals increased. This adsorption sequence was not observed with alkali metal nitrate and alkali metal chloride mobile phases. A brief discussion concerning the effect on RF values of the solvent cations and the adsorption mechanism is included; also presented are typical chromatograms for the separation of multi component mixtures containing adjacent lanthanides. PMID- 11495059 TI - Quinalizarin anchored on Amberlite XAD-2. A new matrix for solid-phase extraction of metal ions for flame atomic absorption spectrometric determination. AB - Amberlite XAD-2 has been functionalized by coupling it to quinalizarin [1,2,5,8 tetrahydroxyanthraquinone] by means of an -N = N- spacer. Elemental analysis, thermogravimetric analysis, and infrared spectra were used to characterize the resulting new polymer matrix. The matrix has been used to preconcentrate Cu(II), Cd(II), Co(II), Pb(II), Zn(II), and Mn(II) before their determination by flame atomic absorption spectrometry (FAAS). UO2(II) has been preconcentrated for fluorimetric determination. The optimum pH values for maximum adsorption of the metals are between 5.0 and 7.0. All these metal ions are desorbed (recovery 91 99%) with 4 mol L(-1) HNO3. The adsorptive capacity of the resin was found to be in the range 0.94-5.28 mg metal g(-1) resin and loading half-life (t1/2) between 5.3 and 15.0 min. The effects of NaF, NaCl, NaNO3, Na2SO4, Na3PO4, Ca(II), and Mg(II) on the adsorption of these metal ions (0.2 microg mL(-1)) are reported. The lower limits of detection for these metal ions are between 1 and 15.0 microg L(-1). After enrichment on this matrix flame AAS has been used to determine these metal ions (except the uranyl ion) in river water samples (RSD < or = 6.5%); fluorimetry was used to determine uranyl ion in well water samples (RSD < or = 6.3%). Cobalt from pharmaceutical vitamin tablets was preconcentrated by use of this chelating resin and estimated by FAAS (RSD approximately 4%). PMID- 11495060 TI - PbO2-based graphite-epoxy electrode for potentiometric determination of acids and bases in aqueous and aqueous-ethanolic media. AB - The construction and analytical evaluation of a PbO2-based graphite-epoxy electrode sensitive to H3O+, based on incorporation of lead(IV) oxide in a graphite-epoxy matrix, are described. The data obtained from a variety of acid base titrations in aqueous and aqueous-ethanolic media were compared with those obtained by use of a glass electrode under the same conditions. The proposed electrode provides a linear response in the pH range from 1 to 11 with a slope of -58.7+/-0.3 mV pH(-1) and -60.8+/-0.2 mV pH(-1) in aqueous and ethanolic media, respectively. The response time was less than 15 s and the lifetime of the electrode was at least eight months (ca. 5000 determinations) and its performance is good in pH determination and end-point detection in potentiometric acid-base titrations in both aqueous and aqueous-ethanolic media. PMID- 11495061 TI - Calibration graphs for steels by IR laser ablation inductively coupled plasma atomic emission spectrometry. AB - Infrared laser ablation (IRLA) was studied as a sample-introduction technique for the analysis of steels by inductively coupled plasma atomic emission spectrometry (ICP-AES). A comparison of two IRLA-ICP-AES systems based on Q-switched nanosecond Nd: YAG lasers was performed. The beam of the LINA-Spark atomizer (LSA Sarl, Cully, Switzerland) based on the Surelite 1-20 laser (Continuum, USA) was moved along a circle. A Perkin-Elmer Optima 3000 DV ICP system was used both with lateral and axial viewing modes. A laboratory-made ablation system based on the Brilliant laser (Quantel) was coupled to a Jobin-Yvon 170 Ultrace ICP (lateral viewing, polychromator part employed). A sample was rotated along a circle during ablation. Linearity of calibration plots was verified at least up to 19% Cr and 12% Ni without internal standardization for both LA-ICP-AES systems. Other elements examined were Mo up to 3%, Mn up 1.5%, Si up to 1.7%, and Cu up to 0.15%. The reproducibility was in the range 5-1 %RSD for a mass percentage 0.5 20% of steel constituents. The relative uncertainty of the centroids of the calibration lines was in the range from +/- 4% to +/- 12% for Cr, Ni, Mn, Mo, and Si, and from +/- 8% to +/- 19% for Cu. The lowest determinable quantities were calculated for calibration dependencies. Performances of both the IR-LA-ICP-AES were comparable. PMID- 11495062 TI - A quartz crystal microbalance sensor for the detection of formic acid vapors. AB - A new quartz crystal microbalance sensor is developed to determine formic acid at low concentrations. Four previously selected polymers with acid-base characteristics were tested as possible coatings. Polyoxyethylene bis [amine] presented the best results. The sensor is rapid, sensitive [0.67 Hz/(mg/m3)], and reversible at low concentrations. The detection limit for formic acid (7.2 mg/m3) is comparable with the short term exposure limit and the threshold limit values. It presents a fast mechanical response to pressure changes, so that it can be quickly used in different environments and situations. The sensor also shows a good stability in a temperature range typical of work atmospheres (16-36 degrees C). It has a wide linear range (7.2-911.2 mg/m3) and a long useful time. It is also applicable to other low molecular mass carboxylic acids such as acetic acid. PMID- 11495063 TI - Determination of inorganic acids by ion chromatography with n tetradecylphosphocholine (zwitterionic surfactant) as the stationary phase and pure water as the mobile phase. AB - A new ion chromatographic (IC) system, in which n-tetradecylphosphocholine (TDPC, a phosphobetaine type of zwitterionic surfactant) was used as the stationary phase, pure water as the mobile phase, and conductivity as the method of detection, has been developed for the determination of inorganic acids. Five model acids, HCl, HNO3, HClO4, H2SO4, and H3PO4, were separated to baseline and eluted in the order H3PO4 > HCl > HNO3 > H2SO4 > HClO4. When peak areas were plotted against the concentrations of the acids in samples, linear calibration curves were obtained. Ultimate determination limits were approximately 1 mmol L( 1), but the discrimination of the method between solutions of different concentration was better than 10 micromol L(-1) for those model analytes. Salts of divalent cations could also be separated, but they were eluted faster than the acids. No separation was observed for the salts of monovalent cations. This newly proposed approach is applicable to the simultaneous determination of the inorganic acids (produced by reactions of NOx, SOx, and HCl with water) in aerosols. PMID- 11495064 TI - High performance liquid chromatographic determination of diazinon, permethrin, DEET (N, N-diethyl-m-toluamide), and their metabolites in rat plasma and urine. AB - A rapid method was developed for the analysis of the insecticide (A) diazinon (O,O-diethyl O-2-isopropyl-6-methylpyridimidinyl) phosphorothioate, its metabolites (B) diazoxon (O,O-diethyl O-2-isopropyl-6-methylpyridimidinyl) phosphate, and (C) 2-isopropyl-6-methyl-4-pyrimidinol, the insecticide (D) permethrin [3-(2,2-dichloro-ethenyl)-2,2-dimethylcyclopropanecarboxylic acid (3 phenoxyphenyl)methylester], its metabolites (E) m-phenoxybenzyl alcohol, and (F) m-phenoxybenzoic acid, the insect repellent (G) DEET (N,N-diethyl-m-toluamide), and its metabolites (H) m-toluamide and (I) m-toluic acid in rat plasma and urine. The method is based on using C18 Sep-Pak cartridges (Waters Corporation, Milford, Mass., U.S.A.) for solid phase extraction and high performance liquid chromatography with a reversed phase C18 column, and absorbance detection at 230 nm for compounds A, B, and C, and at 210 nm for compounds D-I. The compounds were separated using a gradient from 1% to 99% acetonitrile in water (pH 3.0) at a flow rate ranging between 1 and 1.7 mL/min in a period of 17 min. The limits of detection were ranged between 20 and 100 ng/mL, while limits of quantification were 80-200 ng/mL. The relationship between peak areas and concentration was linear over a range of 100-1000 ng/mL. This method was applied to determine the above insecticides and their metabolites following dermal administration in rats. PMID- 11495065 TI - Determination of ultratrace dissolved arsenite in water--selective coprecipitation in the field combined with HGAFS and ICP-MS measurement in the laboratory. AB - Because stabilization of arsenite in water samples during transit and storage is troublesome, this work deals with a method to prevent this by on-site selective coprecipitation of arsenite with dibenzyldithiocarbamate and recovery of the coprecipitate by filtration through a 0.45-microm membrane filter. In the laboratory arsenic on the filter is quantitatively released by oxidation of arsenite to arsenate with H2O2 (6%) in alkaline medium (8 mmol L(-1) NaOH) at elevated temperature (85 degrees C) for 30 min followed by ultratrace determination by routine HGAFS and ICP-MS. It is shown that arsenate contamination of the coprecipitate is so low that arsenate concentrations three orders of magnitude higher than the arsenite concentration do not interfere; this is essential, because arsenate is usually the dominant arsenic species in water. Because significant preconcentration can be achieved in the solution obtained from the leached filter (normally a factor 20 but easily increased to 100) very low detection limits can be obtained (only limited by the purity of the materials and the cleanliness of working); a realistic limit of determination is 0.01 microg L(-1) arsenite. The procedure was used for the determination of arsenite in two ground waters from an ash depository site in the Salek valley (Slovenia). The matrix contained some elements at very high levels but this did not impair the efficiency of arsenite coprecipitation. The results obtained by use of HGAFS and ICP-MS were not significantly different at the 5% level for sub-microg L(-1) arsenite concentrations. PMID- 11495066 TI - Heterogeneity and liability of Pb(II) complexation by humic substances: practical interpretation tools. AB - The complexation of Pb(II) by natural organic matter (NOM) is better described by taking into account the dependence of the strength of binding on metal loading conditions. The utility of a linear differential equilibrium function for interpretation of metal ion binding data is demonstrated. This approach considers the binding intensity (log K*) as a function of metal ion loading (theta = bound metal/binding site concentration). Three methods for calculating this function are presented: -direct calculation from metal titration curves, -direct calculation from polarograms, and -compilation of data derived from interpretation of complexation in terms of one- or two- binding sites (e.g. Scatchard analysis), i.e. Cc (complexation capacity = effective site concentration)-K pairs. Heterogeneity also impacts on the apparent lability of complexes; complexes formed at the lowest metal loadings are the least labile. PMID- 11495067 TI - Metal distribution and binding in balneological peats and their aqueous extracts. AB - Binding of metals in typical bath peat samples ("Grosses Gifhomer Moor", Sassenburg/North Germany) and their aqueous extracts was characterized by means of a multi-method approach. For that purpose a sequential extraction procedure based on peat-filled chromatography columns was developed. Water-soluble metal and DOM (dissolved organic matter) fractions were subdivided by use of a stepwise increased pH gradient (pH 3.8-5), finally by the chelator EDTA and 0.1 mol L(-1) hydrochloric acid. Metal fractions very strongly bound to peat were assessed by an aqua regia extraction. Metal determinations required were performed by atomic spectrometry methods (AAS, ICP-OES, and TXRF). The metal and DOM concentrations in the peat extracts varied significantly, depending on the natural variety of the peat matter under study (e.g., Al: 25-674, Cd: 0.05-0.2, Cu: 5-15.4, Fe: 77 1785, Mn: 21-505, Ni: 2-33, Pb: < 1, Zn: 9-715 (microg L(-1)); Na: 8-45, K: 1.3 14.9, Ca: 2-51, Mg: 1.1-7.9 (mg L(-1)); 26-73 mg L(-1) DOC). An increase of the pH increased the DOC (dissolved organic carbon) of the peat extracts, but hardly the concentration of heavy metals. The latter could only be re-mobilized by EDTA and dilute hydrochloric acid. Additional investigations of the peat extracts using tangential-flow ultrafiltration revealed that the heavy metals extracted at pH < 4 were predominantly dissociated. At higher pH (pH > 4.5) they were preferentially bound to macromolecular DOM. Moreover, using multistage ultrafiltration the size distribution of the DOM and their metal species was assessed. PMID- 11495068 TI - Use of zwitterionic micelles in the eluent II: a new approach to ion chromatographic analysis of inorganic cations in biological fluids with direct sample injection. AB - A new ion chromatographic (IC) technique has been developed for the determination of inorganic cations in biological fluids with direct sample injection. This involved the use of a mixed zwitterionic-micelle/electrolyte solution as an eluent. The proteins in the sample became bound to the zwitterionic micelles in the eluent and were thus eliminated from the column. The cations were separated by cation exchange. This method is ideal for the online, simultaneous determination of common inorganic cations (Na+, NH4+, K+, Mg2+, and Ca2+) in urine and serum samples. Such an application was demonstrated experimentally. Non suppressed conductivity was used for analyte detection. The detection limits obtained using this IC system were 2.94, 5.22, 34.9, 32.6, and 56.7 microg/L for Na+, NH4+, K+, Mg2+, and Ca2+, respectively. PMID- 11495069 TI - Determination of inorganic anions in human saliva by zwitterionic micellar capillary electrophoresis. AB - Capillary electrophoresis (CE) using sulfobetaine-type zwitterionic micelles as the background electrolyte (BGE) has been used to determine inorganic anions in human saliva. The zwitterionic micelles resulted in unique migration behavior for the separation of inorganic anions. They also prevented adsorption of proteins on the inner wall of the capillary. These properties of the zwitterionic micelles enabled the direct determination of inorganic anions in human saliva. Three species of inorganic anions, NO2-, NO3-, and SCN-, were found in real samples and the analysis was achieved within 3 min. Direct UV-absorption was used as the detection method and the detection limits for these anions were 2.0, 1.0, and 5.0 micromol L(-1), respectively (0.09, 0.06, and 0.30 microg mL(-1)). PMID- 11495070 TI - Potentiometric stripping analysis (PSA) for monitoring of antimony in samples of vegetation from a mining area. AB - A potentiometric stripping analysis (PSA) method has been developed and checked for the fast and reliable determination of antimony in vegetation samples of Cistus ladanifer from a mining area in Badajoz, Southwest Spain. The method, modified from previous PSA methods for Sb in environmental samples, is based on dry ashing of the homogenized leaves, dissolution in hydrochloric acid, and PSA analysis on a mercury film plated on to a glassy carbon disk electrode. The influence of experimental variables such as the deposition potential, the deposition time, the signal stability and the calibration parameters, has been investigated. The method has been compared with an independent technique (instrumental neutron activation analysis) by analysis of standards and reference materials and comparison of the results. As a result of automation of the PSA equipment, the proposed method enables unattended analysis of 20 digested samples in a total time of 2 h, thus providing a useful tool for Sb monitoring of a large number of samples. PMID- 11495071 TI - Cardiovascular dysfunction in cirrhosis. Pathophysiological evidence of a cirrhotic cardiomyopathy. PMID- 11495072 TI - Patient self-assessment of test-day symptoms in 24-h pH-metry for suspected gastroesophageal reflux disease. AB - BACKGROUND: Marked daily fluctuations may occur in the pattern and degree of gastroesophageal reflux (GOR) and in patients' symptoms. The aim of this study was to observe how patients' self-assessment of their symptoms on the day of a 24 h pH test correlates with the likely outcome of the test and the potential value in repeating it. METHODS: 367 patients with symptoms suggestive of GOR underwent 24-h pH tests. Fifty-eight patients had repeat studies. Patients assessed the severity of their test-day symptoms as 'better than typical', 'typical' or 'worse than typical'. RESULTS: A 'typical' or 'worse than typical' day was more likely to produce an abnormal test result (P < 0.0001). A normal first test on a 'better than typical' day was more likely to be followed by an abnormal second test than a normal first test on a 'typical' or 'worse than typical' day (55% versus 22%; P = 0.025). The symptom index score, the total acid exposure time on the first test and the presence of oesophagitis were not associated with an abnormal second test (P not significant). CONCLUSIONS: Patients' self-assessment of the severity of their test-day symptoms should be included in the interpretation of 24-h pH tests for suspected GOR. Patients with a normal pH test on a 'better than typical day warrant a repeat test. PMID- 11495073 TI - Psychological intervention influences the outcome of laparoscopic antireflux surgery in patients with stress-related symptoms of gastroesophageal reflux disease. AB - BACKGROUND: Psychological aspects such as stress, emotions, illness behaviour or personality are known to affect the severity of symptoms of gastroesophageal reflux disease (GERD) and can influence medical outcome in some patients. The aim of the present study was to evaluate the efficacy of psychological intervention within routine surgical care on the surgical outcome of laparoscopic antireflux surgery (LARS) in patients with stress-related GERD symptoms during a 1-year follow-up. METHODS: Out of a sample of 196 consecutive patients who required LARS (Nissen fundoplication), a group of 89 (45%) believed that stress was a factor in the cause of their symptoms (stress-related versus stress-unrelated GERD patients). Patients with stress-related symptoms were randomly assigned to the psychological intervention (PI group; n = 42) or control group with routine surgical care (RC group; n =42). Five patients were excluded from the study. Assessments of surgical outcome were: objective clinical data such as DeMeester score or lower oesophageal sphincter pressure, Gastrointestinal Quality-of-Life Index (GIQLI), evaluation of potential side effects such as subjective degree of dysphagia, general impairment as a result of LARS, and patient satisfaction with surgery. RESULTS: There were no significant differences in objective clinical data between the different treatment groups before and after surgery. Before surgery, patients with stress-related symptoms had a lower GIQLI and an increased spectrum of gastrointestinal (GI) symptoms compared with patients without stress related symptoms. A significant impact (P < 0.05-0.01) of psychological intervention on quality-of-life data, especially in GI symptoms, degree of dysphagia and general impairment, could be calculated after surgery. No differences in satisfaction with therapy were detectable. Comparing outcome, no significant differences between patients without stress-related GERD symptoms and the PI group were found. Generally, quality-of-life data in all patients improved significantly and patient satisfaction was excellent or good in 98.9% one year after surgery. In two patients a laparoscopic refundoplication was necessary because of a 'slipping Nissen'. CONCLUSIONS: These findings indicate that there is no impact of psychological intervention on objective clinical data. Patients with stress-related GERD symptoms profit significantly from psychological intervention in patient-related factors of surgical outcome such as quality of life or degree of several aspects such as dysphagia and general impairment. Generally, LARS in patients with stress-related GERD symptoms is an effective and safe procedure which improves quality of life with fewer side effects. Psychological intervention reduces non GERD-related GI symptoms and makes the outcome comparable to the outcome of patients without stress-related symptoms. We therefore suggest that surgical treatment alone in patients with stress-related GERD symptoms is incomplete and that psychological intervention can optimize surgical outcome in these patients. PMID- 11495074 TI - High-dose proton-pump inhibitors as a diagnostic test of gastro-oesophageal reflux disease in endoscopic-negative patients. AB - BACKGROUND: To evaluate a high dose of a proton-pump inhibitor as a diagnostic test in endoscopy-negative patients presenting with symptoms indicating gastro oesophageal reflux disease. METHODS: 64 patients were studied in a prospective, randomized, double-blind study, using a cross-over design. After a run-in period with the diary registration of basic GORD symptoms and recording of the consumption of antacid tablets, the patients were given either 60 mg of lansoprazole once daily or placebo in randomized order. Symptoms were recorded, as well as antacid tablets taken in order to relieve pain. GORD was determined by 24-h oesophageal pH monitoring. The test was considered positive when consumption of antacid tablets was reduced > or = 75% compared to pretreatment. RESULTS: In the GORD group, 29 (85%) tested positive during active treatment compared to 3 (9%) when on placebo. Corresponding figures for the non-GORD patients were 50% and 27%, giving a test sensitivity and specificity of 85% and 73%, respectively. During active treatment, VAS scores for acid regurgitation, heartburn and over all were significantly lowered in GORD patients, compared to heartburn only in the non-GORD group. CONCLUSIONS: 60 mg lansoprazole once daily for 5 days is an easy to use method for diagnosing GORD in endoscopy-negative patients. Using 24-h oesophageal pH monitoring as the reference method, the sensitivity was relatively high, while the specificity was lower. Further studies are needed to determine how a PPI could be used as a diagnostic test in GORD. PMID- 11495075 TI - Betaine-palmitate reduces acetylsalicylic acid-induced gastric damage in rats. AB - BACKGROUND: Acetylsalicylic acid (ASA)-induced gastric injury is reduced when ASA is administered along with phosphatidylcholine. The hydrolysis of endogenous phosphatidylcholine leads to the production of betaine, which may participate in the maintenance of cellular homeostasis. The present aims were to investigate the effects of exogenous betaine and its palmitic acid complex (betaine-palmitate) in the protection of the gastric mucosa in ASA-induced subacute damage. METHODS: Repeated doses of ASA were given intragastrically to male Wistar rats. Control rats were given vehicle only, while treated animals were challenged with ASA or with ASA along with betaine, palmitic acid or betaine-palmitate. The gastric mucosa was examined after 3 days and the nature of any microscopic mucosal injury was assessed by histology. The extent of macroscopic damage, changes in permeability (assessed by Evans blue method) and tissue ATP concentrations were determined in separate series. RESULTS: ASA induced a significant fall in the ATP content of the mucosa, which was not affected by the other drugs used in the study. However, the ASA-induced mucosal permeability increase could be completely reversed by betaine-palmitate supplementation. The extent of severity of the macroscopic and microscopic lesions was 33% and 2.45, respectively, for ASA, as compared with 15% and 2.2 for betaine, 14% and 1.9 for palmitic acid and 3% and 1.4 for betaine-palmitate. CONCLUSIONS: Betaine-palmitate affords a significant protective effect against ASA-induced injury, without influencing the ATP synthesis, and this suggests that the defence is due to its ability to prevent secondary damage. PMID- 11495076 TI - Impact of non-steroidal anti-inflammatory drug and aspirin use on the prevalence of dyspepsia and uncomplicated peptic ulcer disease. AB - BACKGROUND: Non-steroidal anti-inflammatory drug and aspirin (here collectively called NSAIDs) use is the second most common aetiologic factor for peptic ulcer disease and a major factor for peptic ulcer complications. The role of NSAIDs in the pathogenesis of uncomplicated peptic ulcer is less well understood and the interaction between NSAIDs and Helicobacter pylori infection on ulcer development is controversial. The aim of the present study was to examine the role of NSAIDs in the occurrence and clinical features of uncomplicated peptic ulcer disease. METHODS: A total of 1091 consecutive patients referred for open-access upper gastrointestinal endoscopy by general practitioners (GPs) were enrolled. The use of NSAIDs was gathered from a structured questionnaire completed by the patients and from patient files by GPs. The exclusion criteria were previous H. pylori eradication and gastric surgery, as well as symptoms and/or signs suggestive of acute gastrointestinal bleeding. RESULTS: Of the whole study group (n = 1091), 76 (7%) patients had a peptic ulcer. Thirty patients had an NSAID-use-associated peptic ulcer and 46 patients a non-NSAID-use peptic ulcer. Of patients with chronic gastritis (n = 599), 71% were H. pylori-positive and 108 used NSAIDs. Of those with chronic gastritis, 23 had an NSAID-use-associated peptic ulcer and 38 a non-NSAID ulcer. Of patients with normal gastric histology (n = 492), 75 patients used NSAIDs, 7 had an NSAID ulcer and 8 a non-NSAID ulcer. The only independent risk factor for peptic ulcer in patients using NSAIDs was H. pylori infection (odds ratio (OR) 3.1, 95% confidence interval (CI) 1.3-7.3), whereas dyspepsia (OR 1.0, 95% CI 0.4-2.4), male sex (OR 1.4, 95% CI 0.6-3.4), age (OR 1.0 per decade, 95% CI 0.8-1.3) and anaemia (OR 2.9, 95% CI 0.9-8.7) were not risk factors. In patients not using NSAIDs, independent risk factors for peptic ulcer were dyspepsia (OR 4.3, 95% CI 2.1-8.8), male sex (OR 2.0, 95% CI 1.1-2.8), age (OR 1.2 per decade, 95% CI 1.0-1.5), anaemia (OR 6.2, 95% CI 2.6-14.9) and H. pylori infection (OR 7.5, 95% CI 3.4-16.6). When comparing patients using NSAIDs or not, the OR of patients on NSAIDs for peptic ulcer was 2.7 (95% CI 1.5-5.0) among patients with chronic H. pylori gastritis (n = 424) and 5.3 (95% CI 1.8 15.0) among patients with normal gastric mucosa (n = 492). CONCLUSIONS: The use of NSAIDs increases the risk of peptic ulcer 3- and 5-fold in H. pylori-positive and H. pylori-negative patients, respectively. Dyspepsia is a poor predictor of peptic ulcer among patients using NSAIDs, and serologic H. pylori testing and treatment for chronic NSAID users is recommended. PMID- 11495077 TI - Refinement of the 14C-urea breath test for detection of Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori is recognized as the main etiological factor for chronic gastritis, peptic ulcer and possibly also gastric malignancies. A 14C urea breath test was successively refined to correctly diagnose the presence of H. pylori. METHODS: After intake of a 14C-urea cocktail, 14CO2 in breath was trapped in benzethoniumhydroxide/ethanol. Serology by ELISA, followed by Western blot, was used as reference method for confirmation of presence or absence of H. pylori. RESULTS: Breath measurements at 10 and 20 min were determined to be optimal for diagnostic purposes. With 2.5 microCi of 14C-urea, a sensitivity of 86% and specificity of 100% was obtained for the 10-min values. Corresponding data for the 20-min values were 86% and 99%, respectively. When 50 mmol L(-1) citric acid was added to the 2.5 microCi 14C-urea cocktail, separation between positive and negative results became more distinct along with improved sensitivity of 91% and specificity of 100%. As the isotope amount was reduced to 1.0 microCi in the citric acid-containing cocktail, a sensitivity of 93% and specificity of 100% were obtained at both 10 and 20 min. CONCLUSION: Diagnostic urea breath test has a high concordance with H. pylori serology, combining ELISA and Western blot. Stepwise refinements proved the optimal urea breath test cocktail to include 1.0 microCi 14C-urea in citric acid solution with breath measurement at 10 min. PMID- 11495078 TI - Sensitivity and distensibility of the rectum and sigmoid colon in patients with irritable bowel syndrome. AB - BACKGROUND: Hyperalgesia to visceral stimuli is a biological marker of the irritable bowel syndrome (IBS). Abnormal pain processing is probably of most importance, but biomechanical abnormalities of the gut wall may also contribute to the findings. In the current study, we investigated the sensation of the gut to electrical stimuli as well as the distensibility of the rectum and sigmoid colon in IBS patients and a control group. METHODS: Nine patients with IBS and 11 controls entered the study. The pain threshold to electrical stimuli at the rectosigmoid junction was determined with bipolar electrodes integrated on the biopsy forceps for the endoscope. Subsequently, controlled distensions of the sigmoid colon and rectum were performed with a balloon integrated on a probe for impedance planimetry, providing the possibility to measure the cross-sectional area (CSA), wall tension and strain to different pressures together with the sensation ratings. RESULTS: The pain detection thresholds to electrical stimuli at the rectosigmoid junction were 12.5 (range 7-39) mA in controls and 7.5 (range 0.75-12) mA in IBS patients (P = 0.03). The calculated pressures at the pain detection threshold in the sigmoid colon were lower in the IBS patients (31.5 (range 5-58) versus 5 cm (range 5-25) water; P = 0.03), otherwise no differences were seen in sensation rating to the different distension pressures. The CSA was slightly higher in controls to the different pressures, whereas no differences between the groups were seen in strain and tension of the rectum and sigmoid colon. CONCLUSION: The visceral hypersensitivity in IBS seems to be related to alterations in the nervous system rather than biomechanical parameters such as the tension and strain of the gut wall. Treatment of pain in IBS should therefore be based on drugs with documented action on the nociceptive pathways in the central nervous system. PMID- 11495079 TI - Abdominal symptoms and anorectal function in health and irritable bowel syndrome. AB - BACKGROUND: It is unclear how the quality and quantity of abdominal symptoms and anorectal function differ between irritable bowel syndrome (IBS) patients and healthy controls, and whether different anorectal function in patients is associated with abdominal symptoms in IBS. METHODS: Fifty-two outpatients with IBS and 12 healthy controls kept daily symptom records over 1 week. At the end of the week, anorectal function was assessed by manovolumetry before and after a standard fatty meal. Patients were divided into symptom and manovolumetric subgroups using a cluster analysis and also into those below (hypersensitive) and those within (normosensitive) the 95% confidence interval of the controls' mean for maximal tolerable distension (MTD). RESULTS: Regardless of subgroup, the patients were distinguished from the controls by pain, bloating, straining and incomplete evacuation. Compared with controls, MTD was lower in the pain/bloating subgroup characterized by considerable pain and the bowel habit subgroup characterized by hard stools, variable stool consistency and heavily disturbed stool passage. Preprandial rectal hypersensitivity was highly prevalent in this bowel habit subgroup. No similar association with the pain/bloating subgroup was found. Patients and controls showed a significant and similar postprandial decrease in MTD. CONCLUSIONS: IBS is distinguished from health by pain, bloating, straining and a feeling of incomplete evacuation. Baseline rectal hypersensitivity is associated with constipation-like bowel habit. Increased rectal sensitivity after a meal and/or preceding distension is a normal reaction unimportant in the genesis of symptoms in IBS. PMID- 11495080 TI - Gastric IgA in cystic fibrosis in relation to the migrating motor complex. AB - BACKGROUND: Gastrointestinal symptoms in cystic fibrosis are frequent, but little is known about the underlying pathophysiology. Mucosal secretion of IgA is important for the immunologic function in the human gastrointestinal tract but has not been studied in cystic fibrosis. The aim of this study was to quantify the release of IgA by the gastric mucosa in relation to interdigestive motor activity in patients with cystic fibrosis with different genotypes. METHODS: The study included 7 healthy adult volunteers and 10 adult patients with cystic fibrosis, all Helicobacter pylori-negative. All patients had pathological sweat tests and clinical symptoms and signs of cystic fibrosis. All but one were colonized with Pseudomonas aeruginosa. Three patients were pancreatic sufficient. The investigation was performed using intragastric perfusion and gastroduodenal manometry. RESULTS: During the investigation, 8 of 10 patients with cystic fibrosis showed the characteristic pattern of interdigestive motility. The patients had significantly lower levels of gastric IgA compared to healthy subjects during phases II and III of migrating motor complex, median (range) 120 (67-442) and 36 (6-299) microg/5 min. 382 (40-1176) and 56 (4-398) (P = 0.03 and P = 0.04), respectively. Only one patient with genotype R668C/unknown showed IgA levels within the normal range. There was no correlation to gastric presence of duodenogastric reflux markers. CONCLUSION: The interdigestive motility pattern was normal in most patients with cystic fibrosis. The low levels of IgA released from the gastric mucosa in the patients might indicate a defective gastric transmucosal IgA transport in cystic fibrosis. PMID- 11495081 TI - Anti-transglutaminase IgA ELISA: clinical potential and drawbacks in celiac disease diagnosis. AB - BACKGROUND: Since the identification of tissue transglutaminase (tTG) as the antigen for the anti-endomysial antibodies (EMA), several antigen-specific immunoassays have been reported for celiac disease (CD) screening. A first objective was to evaluate the suitability for CD screening of three different IgA tTG ELISAs, two of them based on guinea pig liver tTG (gp-tTG) (an in-house ELISA with a partially purified extract and a commercial ELISA with purified gp-tTG antigen) and a third recombinant human tTG (rh-tTG) ELISA. The results are compared with EMA and with the final clinical diagnosis. A second objective was to analyze antibody reactivities in those patients with anti-tTG and EMA discrepancies. METHODS: ELISA and EMA tests were used to measure IgA anti-tTG levels in sera from 259 patients (107 had CD and 72 had Type I diabetes mellitus). RESULTS: The purified gp-tTG ELISA was highly sensitive (97.7%) and specific (98.8%) in the detection of CD, almost equaling EMA. Rh-tTG ELISA did not improved the sensitivity of EMA, but its specificity was slightly superior. Immunoblot analysis with partially purified gp-tTG extract, the antigen most frequently used for anti-tTG detection, showed that the majority of false positives were due to IgA reactivities to contaminant proteins present in the liver antigenic extract. This low specificity was particularly problematic in diabetics. CONCLUSION: Purified tTG ELISAs, either with purified guinea pig liver or recombinant human antigens, can be used as quantitative and observer independent alternatives to the traditional and time-consuming EMA in the screening of CD. PMID- 11495082 TI - Coordinate upregulation of interleukin-8 and growth-related gene product-alpha is present in the colonic mucosa of inflammatory bowel. AB - BACKGROUND: Although alpha-chemokines, such as interleukin (IL)-8 and epithelial neutrophil-activating peptide 78, are implicated in the pathogenesis of inflammatory bowel disease (IBD), little information is currently available on the expression and cellular source of growth-related gene product-alpha (GROalpha) and its functional relationship to other alpha-chemokines in the intestinal mucosa of patients with IBD. METHODS: The contents of IL-8 and GROalpha in organ cultures, the expression of IL-8 and GROalpha mRNA, and the modulatory effects of inflammatory mediators on IL-8 and GROalpha-producing cells were examined using colonic mucosal tissues. In vitro stimulatory effects of IL-8 and GROalpha on neutrophils were investigated in terms of chemotactic migration and superoxide anion generation. RESULTS: The contents of IL-8 and GROalpha in organ cultures were elevated in patients with IBD, especially in those with active ulcerative colitis (UC). Both IL-8 and GROalpha contents increased according to an increase in histological disease activity in patients with UC, but not in those with Crohn disease. In contrast, no significant correlation was found between the contents of these alpha-chemokines and clinical disease activity. In situ hybridization detected increased expression of IL-8 and GROalpha mRNA in macrophages, pericrypt myofibroblasts, and the epithelium of tissue specimens with active lesions of IBD. The secretion of IL-8 and GROalpha from macrophages and myofibroblasts obtained from control patients was upregulated by inflammatory cytokines and bacterial products. The concentrations of recombinant (r)-IL-8, which covered the levels of activity detected in individual organ cultures or cell cultures of fractionated mucosal cells, could induce chemotactic migration and superoxide anion generation in neutrophils in vitro, and r-GROalpha had synergistic effects on r-IL-8-induced neutrophil activation. CONCLUSIONS: A coordinate upregulation of IL-8 and GROalpha may be involved in the tissue injury in patients with IBD through their stimulatory effects on neutrophils. PMID- 11495083 TI - Enhanced cyclooxygenase-2 expression in sporadic and familial adenomatous polyposis of the human colon. AB - BACKGROUND: The cyclooxygenase (COX) enzymes exist in two related but unique isoforms (COX-1 and COX-2) and catalyze the formation of prostaglandins (PGs). COX-1 is constitutively expressed, and is responsible for the synthesis of PGs necessary for gastroprotection and normal renal function. The COX-2 isoform is important in a variety of pathophysiological conditions such as inflammation and tumorigenesis. Numerous studies report that regular use of non-steroidal anti inflammatory drugs (NSAIDs) can decrease the incidence of some tumor types, including gastrointestinal polyposis. METHODS: In this study, we evaluated COX-1 and COX-2 expression in 30 polyps collected from 10 patients with familial adenomatous polyposis (FAP) and in 18 polyps collected from 18 patients with sporadic adenomatous polyposis (SAP) using COX-1 or COX-2 isoform-specific antibodies. All tissues were formalin-fixed and paraffin-embedded. Immunoreactivity was detected using tyramide signal amplification and evaluated utilizing an immunohistochemical scoring system. RESULTS: COX-2 was minimally detected in the distant non-neoplastic epithelium, which also served as an internal negative control. In comparison, all polyps collected from SAP or FAP patients overexpressed COX-2 in the neoplastic epithelial cells (P < or = 0.002). Additionally, pronounced COX-2 expression was observed in the stromal cells underlying and adjacent to adenomatous lesions. COX-1 immunoreactivity was weak to mild throughout each tissue evaluated and did not change in the neoplastic or stromal cells of the polyps. CONCLUSIONS: COX-2 expression is upregulated in the adenomatous epithelium of SAP and FAP, while the COX-1 isoform appears to be constitutively expressed at low levels in both neoplastic and non-neoplastic regions. The differential expression of COX-1 and COX-2 in these neoplasms suggests that COX-2 rather than COX-1 may play a role in adenoma formation and/or growth in cases of SAP and FAP in humans. PMID- 11495084 TI - Cost-effectiveness of interferon alfa-2b with and without ribavirin as therapy for chronic hepatitis C in Sweden. AB - BACKGROUND: Recent trials have shown that treatment with a combination of interferon alfa-2b and ribavirin results in sustained loss of detectable hepatitis C-virus (HCV) RNA in a higher proportion of patients than treatment with interferon alone. Combination therapy, however, is two to three times as expensive as monotherapy. METHODS: Based on data from recent randomized clinical trials and a previously published decision model, we developed a Markov model to estimate the cost-effectiveness of initial combination therapy with interferon and ribavirin versus interferon alone for previously untreated patients with chronic HCV infection in Sweden. Clinical praxis and quality adjustments were based on expert estimates and costs were gathered from different health care providers in Sweden. RESULTS: Combination therapy for 24 or 48 weeks, compared to interferon alone, prolonged quality adjusted life expectancy by 0.5 to 1.1 years at marginal cost-effectiveness ratios of US$ 1,400 to US$ 6,000 per DQALY (discounted quality-adjusted life-year) for patients with genotype 1. In genotype 1, 48 weeks compared to 24 weeks of combination therapy prolonged quality adjusted life expectancy by 0.6 years at a marginal cost-effectiveness ratio of $US 9,800 per DQALY. For patients with genotype non-1, combination therapy for 24 or 48 weeks, compared to interferon alone, prolonged quality adjusted life expectancy by 2.3 years, with combination therapy for 24 weeks being money saving. The results were robust in sensitivity analyses. CONCLUSION: Combination therapy with interferon and ribavirin increased quality-adjusted life expectancy and was cost-effective for patients with chronic hepatitis C. PMID- 11495085 TI - Serum dipeptidyl peptidase IV (DPP IV, CD26) activity in chronic hepatitis C. AB - BACKGROUND: DPP IV is a cell surface ectoenzyme widely distributed in the human body. It has been implicated in T-cell activation, hepatocyte-extracellular matrix interactions and fibroblast proliferation. Furthermore, upregulated CD26 expression has been found on the surface of human hepatoma cells transfected with hepatitis C virus (HCV) c-DNA. We examined the serum DPP IV activity in a large number of patients with chronic HCV infection in a cross-sectional study. We also investigated whether the activity differs from that in controls and depends upon the response to interferon (IFN) therapy. METHODS: Serum DPP IV activity was measured by microplate-based (Multiskan-Plus-MKII, Labsystem) kinetic assay in 144 patients with chronic HCV infection. Seventy-four out of 144 patients (46 non responders, 28 responders) were formerly treated with interferon. Sixty healthy blood donors served as controls. Gly-Pro-PNA (Bachem, Torrance, USA) was used as substrate. Results are expressed in nmol/ml/min (U/l). Shapiro-Wilk's test, Mann Whitney U test and Spearman rank order correlation were used for statistical analysis. RESULTS: Serum DPP IV activity was significantly higher (mean = 20.89 [s 9.6]) in patients with chronic HCV infection than in healthy controls (12.39 [2.76, P < 10(-5)]). The enzyme activities significantly differed in naive HCV positive patients (22.2 [9.89, P < 10(-5)]) and non-responders (23.28 [9.57, P < 10(-5)]) from that in the healthy controls and also from that in responders (13.69 [4.21]). Correlation was found between DPP IV activity and AST (r = 0.44, P < 10(-5)), ALT (r = 0.44, P < 10(-5)), GGT (r = 0.41, P < 10(-5)) levels. CONCLUSION: Serum DPP IV activity seems to be an indicator of HCV induced liver injury. The activity may reflect the efficacy of interferon therapy. PMID- 11495086 TI - HFE haemochromatosis gene mutations in liver transplant patients. AB - BACKGROUND: The majority of patients with inherited haemochromatosis carry two mutant alleles of the recently discovered HFE gene. Individuals heterozygous for the HFE mutation could be predisposed to end-stage liver disease due to other causes. METHODS: The frequencies of the HFE gene mutations C282Y and H63D were determined in DNA samples obtained from 189 liver transplant patients and 225 healthy Finnish blood donors. RESULTS: 5% of the 189 liver transplant recipients were heterozygotes and 0.5% homozygotes for the C282Y mutation, while 16% were heterozygotes and 0.5% homozygotes for the H63D mutation. These figures were not increased in comparison to controls, of whom 11% were C282Y heterozygotes, 16% H63D heterozygotes and 0.9% H63D homozygotes. Among recipients with acute non-A-E hepatitis (n = 31), the frequency of the H63D allele was higher than in controls (21% versus 9.1%, P < 0.01). Perls' stain for iron in explanted liver specimens was positive in 28% of recipients with alcoholic cirrhosis, 26% of patients with acute non-A-E hepatitis and 14% in the rest of the recipients. The HFE genotypes did not correlate with the iron status. CONCLUSION: Individuals heterozygous for either the C282Y or H63D mutation of the HFE gene are not at increased risk of developing chronic end-stage liver disease. However, subjects heterozygous for the H63D mutation may have an increased risk to develop fulminant non-A-E hepatitis. PMID- 11495087 TI - The HLA-DR3,DQ2 heterozygous genotype is associated with an accelerated progression of primary sclerosing cholangitis. AB - BACKGROUND: An improvement of prognostic models in primary sclerosing cholangitis (PSC) is needed. In particular, inclusion of prognostic markers that are independent of the disease stage would be advantageous. We investigated whether HLA class II genes associated with PSC are also related to disease progression. METHODS: The study included 265 PSC patients from five European countries with a median follow-up of 9.1 years. The end-points were death (n = 38) or liver transplantation (n = 52). Thirty patients developed cholangiocarcinoma during follow-up. RESULTS: The DRB1*03,DQA1*0501, DQB1*02 (i.e. DR3,DQ2) heterozygous genotype was associated with an increased risk of death or liver transplantation (hazard ratio = 1.63; 95% confidence interval (CI) = 1.06-2.52). The presence of a DQ6 encoding haplotype (DQB1*0603 or DQB1*0602) in DR3,DQ2 negative individuals was associated with a reduced risk of death or liver transplantation (hazard ratio = 0.57; 95% CI = 0.36-0.88). There was a trend towards an increased risk of developing cholangiocarcinoma among DR4,DQ8 positive patients, but this did not reach significance (odds ratio = 2.27; 95% CI = 0.78-6.62). CONCLUSION: The DR3,DQ2 heterozygous genotype is associated with a more rapid progression of PSC, whereas HLA-DQ6 is associated with a retarded disease progression. It is possible that the DR4,DQ8 haplotype is related to cholangiocarcinoma development. PMID- 11495088 TI - Multiple hepatic abscesses due to Yersinia enterocolitica infection secondary to primary haemochromatosis. AB - A case of hepatic abscesses due to Yersinia enterocolitica in an immunocompetent male is presented. Re-examination after 3 months showed that the patient had primary haemochromatosis. Treatment with repeated phlebotomies was instituted. Two years after the patient was first admitted to hospital. 17.2 g iron had been removed and all haematological and biochemical parameters had returned to normal. Genetic analysis of the patients' two sons showed that one was positive for the chromosome defect found in primary haemochromatosis; further investigation is under progress. A study of the literature showed that prior to this case only 45 cases of hepatic abscess secondary to Yersinia enterocolitica have been registered. Of the 45 reported cases, 64% had underlying haemochromatosis and 29% had diabetes mellitus. The overall mortality was 31%. Mortality before 1987 was 60% (n = 20) and since 1987 it has been 8% (n = 25). PMID- 11495089 TI - Pernicious anemia and stomach cancer. PMID- 11495090 TI - The impact of posttraumatic stress disorder in military situations. AB - The constant threat to life and gruesome sights and sounds of war take their toll on the soldier psychologically as well as physically. A significant number of war veterans suffer from a wide range of debilitating psychological symptoms that vary in duration. For some the symptoms are transient, while for others profound and prolonged psychological and somatic sequelae manifest in the form of posttraumatic stress disorder (PTSD) and other comorbid conditions. This article reviews current understanding regarding combat stress reaction--often the first indicator of psychological breakdown--and posttraumatic sequelae. The longer-term detrimental consequences of PTSD and the impact of secondary traumatization. reactivation, and delayed-onset PTSD are also addressed. PMID- 11495091 TI - The epidemiology of posttraumatic stress disorder: what is the extent of the problem? AB - Until recently, our understanding of posttraumatic stress disorder (PTSD) relied almost entirely on studies of war veterans and disaster victims. A handful of epidemiologic studies have now been conducted that investigate the natural course of PTSD as it occurs in the general population. Estimates of PTSD prevalence have tended to vary according to the diagnostic criteria used to define the disorder, assessment procedures, sample characteristics, and the definition of qualifying traumatic events. This article reviews key findings from these studies to provide insight into the burden of PTSD in the general population. Possible reasons for the observed difference in lifetime prevalence of PTSD between the sexes (a female-to-male lifetime prevalence ratio of 2:1 is typically reported) and factors thought to be associated with an increased risk for the disorder after exposure to trauma are reviewed. PMID- 11495092 TI - Posttraumatic stress disorder in the general population and in children. AB - Posttraumatic stress disorder (PTSD) is a complex psychopathologic condition that represents a significant challenge to the psychiatric profession. This distressing disorder has been found to affect both adults and children, although the pattern of symptoms in children can differ from that commonly seen in adults. This article presents an overview of the prevalence and incidence of PTSD and discusses factors that may be influential in the development of this disorder following exposure to traumatic events. In addition. the clinical presentation of PTSD in children is reviewed and treatment options for affected children are discussed. PMID- 11495093 TI - Women and traumatic events. AB - Posttraumatic stress disorder (PTSD) gained the status of a psychiatric disorder in 1980, although the syndrome had already been recognized widely for many years. PTSD is distinguished by alternations between reexperiencing of the traumatic event that triggered the PTSD in the first place and avoidance and numbing. Increased arousal (e.g., exaggerated startle reaction) also forms part of the diagnosis. Although the majority of trauma victims recover spontaneously, more than 30% develop persistent PTSD symptoms, with women being twice as likely as men to suffer PTSD. To date, the most studied psychosocial treatments for PTSD are the cognitive-behavioral interventions. Exposure therapy (systematic exposure to the traumatic memory in a safe environment) has been demonstrated to be quite effective with adult women who were sexually or nonsexually assaulted in adulthood as well as with women who were sexually abused in childhood. Supportive counseling does not appear as effective as exposure therapy, but is better than no therapy. PMID- 11495094 TI - Acute posttraumatic stress: nonacceptance of early intervention. AB - Psychological resistance may be of considerable importance in the posttraumatic stress disorder (PTSD) population, considering that researchers in the field of traumatic stress are frequently unsuccessful in achieving high response rates, that many subjects suffering from PTSD never seek help, and that dropouts from therapy are frequent. This article presents data on the main complaints reported in the acute aftermath of an industrial disaster by 246 employees who had been exposed to the disaster. The dominant concerns were symptomatic complaints related to posttraumatic stress reactions rather than external problems. Sleep disturbance, anxiety/fear responses, and physical symptoms were reported by individuals with complaints in the acute phase as most problematic, while irritability and depressive symptoms appeared very infrequently among the reported main complaints. A high specificity and sensitivity were achieved in predicting later PTSD (as defined by DSM-III criteria) by applying early response variables: thus, there were few false-positives and false-negatives. There was a considerable overlap between the PTSD predictors and the main symptom complaints. From a prevention point of view, this should be advantageous, since it would bring the right people to seek help. However, in a significant proportion of the acutely distressed, the reluctance to seek help was motivated by the very symptoms that predicted PTSD. Even a relatively high rate of subjects agreeing to be screened (82.8%) would have lost 42% of those who qualified for a diagnosis of PTSD, and more than half of the subjects with severe outcomes would not have been included. For primary and secondary prevention, the findings suggest that early screening and outreach should be very active. PMID- 11495095 TI - What is posttraumatic stress disorder? AB - Our understanding of posttraumatic stress disorder (PTSD) has increased significantly over the last 2 decades. Although the cause of the condition is usually easy to determine in individual patients, the symptoms of PTSD are diverse and a mixture of psychological processes are involved. This article presents a broad overview of PTSD, including its definition according to DSM-IV and ICD-10 diagnostic criteria, and its clinical course with reference to its association with depression and other mental disorders. The article also briefly reviews the assessment of patients and considers physiologic features such as responses to startle stimuli that appear to be useful in diagnosing PTSD and in differentiating it from other anxiety disorders and depression. Finally, a brief overview of the treatment of PTSD is given, including psychological and biological treatment options. PMID- 11495096 TI - Biology of posttraumatic stress disorder. AB - Most biological findings in posttraumatic stress disorder (PTSD) are compatible with those of the chronic stress response, such as increased corticotropin releasing factor (CRF) concentrations, catecholamine depletion within the central nervous system, and reduced hippocampal volume. However, over the last 10 years, biological observations have been made in PTSD that are different from what has been typically associated with chronic stress, notably certain hypothalamic pituitary-adrenal (HPA) axis findings. In particular, urinary and plasma cortisol levels are considerably lower in PTSD patients than in non-PTSD trauma survivors and normal controls. Furthermore, the circadian pattern of cortisol release from the adrenal glands follows a greater dynamic range in PTSD than in patients with major depression or in normal controls. The reduction in cortisol levels results from an enhanced negative feedback by cortisol, which is secondary to an increased sensitivity of glucocorticoid receptors in target tissues. This HPA axis alteration contrasts with the well-known chronic stress cascade in which CRF release results in erosion of negative feedback and down-regulation of glucocorticoid receptors. Sensitization of the HPA axis is consistent with the clinical picture of hyperreactivity and hyperresponsiveness in PTSD. PMID- 11495097 TI - Investigating the pathogenesis of posttraumatic stress disorder with neuroimaging. AB - Rapidly evolving brain neuroimaging techniques such as magnetic resonance imaging (MRI) and positron emission tomography (PET) are proving fruitful in exploring the pathogenesis and pathophysiology of posttraumatic stress disorder (PTSD). Structural abnormalities in PTSD found with MRI include nonspecific white matter lesions and decreased hippocampal volume. These abnormalities may reflect pretrauma vulnerability to develop PTSD, or they may be a consequence of traumatic exposure, PTSD, and/or PTSD sequelae. Functional neuroimaging symptom provocation and cognitive activation paradigms using PET measurement of regional cerebral blood flow have revealed greater activation of the amygdala and anterior paralimbic structures (which are known to be involved in processing negative emotions such as fear), greater deactivation of Broca's region (motor speech) and other nonlimbic cortical regions, and failure of activation of the cingulate cortex (which possibly plays an inhibitory role) in response to trauma-related stimuli in individuals with PTSD. Functional MRI research has shown the amygdala to be hyperresponsive to fear-related stimuli in this disorder. Research with PET suggests that cortical, notably hippocampal, metabolism is suppressed to a greater extent by pharmacologic stimulation of the noradrenergic system in persons with PTSD. The growth of knowledge concerning the anatomical and neurochemical basis of this important mental disorder will hopefully eventually lead to rational psychological and pharmacologic treatments. PMID- 11495098 TI - Outcomes of posttraumatic stress disorder. AB - The typical reaction to a life-threatening experience is one of distress, anxiety, and fear. This is characteristic of the basic survival instinct; these emotions enhance the individual's memory of the traumatic experience and thus serve to help in the recognition and avoidance of similarly dangerous situations in the future. In a significant minority of individuals, however, the natural reaction to trauma becomes uncontrollably and disastrously intensified, resulting in the symptoms of posttraumatic stress disorder (PTSD). PTSD varies in severity and duration between individuals, often relating to personal characteristics and the nature of the trauma to which a person is subjected. However, several factors, namely, chronicity, impairment, comorbidity, and somatization, are significantly related to and can influence the course of PTSD and subsequent outcome. This article briefly reviews each of these factors. PMID- 11495099 TI - Topiramate in posttraumatic stress disorder: preliminary clinical observations. AB - Posttraumatic stress disorder (PTSD) is a serious and debilitating mental condition that affects a significant proportion of the general population at some time in their lives. To date, however, the U.S. Food and Drug Administration has approved only 1 pharmacologic treatment for this indication. Additional effective therapies are urgently required to control the destructive symptoms experienced by individuals with PTSD. This article reviews the effects of the novel antiepileptic drug topiramate on 3 patients meeting DSM-IV criteria for chronic PTSD. In these previously treatment-refractory patients, topiramate had a marked effect: reducing and even eliminating trauma-related intrusive memories and nightmares and normalizing depressed mood. Adverse events were effectively controlled with careful drug titration and discontinuation of concomitant therapies. These findings, together with observations in more than 30 additional patients (reported elsewhere), suggest that further study of topiramate as a treatment for PTSD is warranted. PMID- 11495100 TI - Infections related to breast implants reported to the Food and Drug Administration, 1977-1997. AB - The FDA has a surveillance system for monitoring adverse events related to medical devices. Infection reports submitted to the FDA by breast implant manufacturers between 1977 and 1997 are characterized. Two cases of death caused by toxic shock syndrome after mammoplasty reported to the FDA are presented. Overall, 1,971 reports with a principal adverse event of infection were reported in this time frame. There was a large increase in the number of reports on infections related to breast implants between 1992 and 1995 due to the publicity and litigation surrounding breast implants. When an organism was identified in the report, the most common organism reported was Staphylococcus sp. Information on the time between the implantation and the onset of the infection or the explantation of the implant was not always reported. However, in reports that did contain this information, there were differences between the length of time to infection onset reported for saline breast implants (earlier) compared to silicone gel breast implants (later). More than half of the reports (56.6%) asserted only that there was an infection and that breast implants were explanted as a result; the remaining reports asserted that infection and other signs, symptoms, or diagnoses had afflicted the patient. PMID- 11495101 TI - The scientific basis for selecting wound-closure techniques. PMID- 11495102 TI - Hazards of powder on surgical and examination gloves: a collective review. AB - This article reviews information on the hazards associated with dusting powders on latex surgical and examination gloves. Dusting powders were first applied to latex gloves to facilitate donning. After 1980, manufacturers devised innovative techniques to manufacture gloves without dusting powders. It has been well documented that the powders on gloves present a health hazard to patients, as well as to operating-room personnel. First, these powders elicit tissue toxicity in every tissue in the body. Second, these powders serve as carriers of latex allergen and may precipitate a life-threatening allergic reaction in sensitized patients. These well-documented hazards of glove powders have caused a growing number of hospitals in the world to abandon the use of examination and surgical gloves coated with powder, and instead to use only powder-free gloves. PMID- 11495103 TI - The scientific basis for selecting surgical sutures. AB - The purpose of this study was to examine the handling properties of synthetic absorbable and monofilament polypropylene sutures made by two different manufacturers and to compare a subjective evaluation by surgeons to the results of standardized biomechanical performance tests. The surgeons' clinical subjective evaluation of the monofilament polypropylene sutures correlated with the results of the biomechanical performance studies, whereas the biomechanical evaluation of the two synthetic absorbable sutures detected notable differences in their performance. These distinct differences in biomechanical performance of the absorbable sutures did not alter their performance in the subjective clinical evaluation. PMID- 11495104 TI - An innovative absorbable coating for the polybutester suture. AB - The polybutester (PBE) suture has been coated with an absorbable polytribolate polymer that is composed of glycolide (9%), epsilon-caprolactone (51%), and poloxamer 188 (40%) to reduce its drag forces. It is the purpose of this study to document the influence of this coating on the biomechanical performance of both PBE sutures and polypropylene (PP) sutures. The performance parameters evaluated were breaking strength, elongation, stiffness, knot security, knot run down, and tissue drag. The breaking strength of PP sutures was remarkably similar to that of coated and uncoated PBE sutures. In size 5/0 PP sutures, the sutures exhibited considerably greater elongation at knot break than did comparably sized PBE sutures. The PBE suture elongated under low loads, but returned to its original length when the load was removed. In contrast, the PP suture elongated irreversibly at high loads, exhibiting creep. Coated and uncoated PBE sutures exhibited less stiffness than PP sutures and exhibited limited memory after removal from the suture package. Coating the PBE suture markedly reduced its drag forces in musculoaponeurotic, colonic, and vascular tissue. Knot security with the coated PBE suture was achieved with only one more throw than with comparably sized uncoated PBE sutures. On the basis of the results of this study, coating the PBE suture represents another major advance in suture performance. PMID- 11495106 TI - Fibrin sealants in microvascular surgery: current status. AB - During the last two decades, advances in fibrin sealant formulation have resulted in its investigational and clinical use in various surgical endeavors, including microvascular surgery. Several investigations have comparatively evaluated fibrin adhesive-enhanced microvascular anastomoses vs. conventional suture repair. The purpose of this review is to summarize the collective documentation on fibrin adhesives in microvascular surgery on the basis of the scientific performance parameters of vessel patency, bursting strength, anastomotic competence, and reendothelialization. In addition, other applications of fibrin sealants and other qualities unique to fibrin adhesives are addressed. PMID- 11495105 TI - Biomechanical performance of new cardiovascular needles. AB - Cardiovascular needles are now being manufactured from new stainless steel alloys containing high concentrations of nickel, Surgalloy and Ethalloy. The purpose of this study was to compare the biomechanical performance of a cardiovascular needle made of Surgalloy with a comparably sized needle made of Ethalloy. The parameters of biomechanical performance included sharpness, maintenance of sharpness, resistance to bending, and ductility. Because the biomechanical performance of these needles was remarkably similar, cardiovascular needles made of either the Surgalloy or Ethalloy alloys are recommended for cardiovascular surgery. PMID- 11495107 TI - Revolutionary advances in skin stapling: a collective review. AB - The purpose of this article is to provide a collective review of the literature on skin staplers. On the basis of this review, the Auto Suture Multifire Premium, with its regular and wide metal staples, is recommended for skin closure. The Auto Suture Multifire Graftac-S is ideally suited for skin-graft fixation, because it delivers an absorbable staple that usually does not have to be removed from the healing graft. Dermal skin closure can now be achieved using the Auto Suture SQS-20, which coapts the cut dermal edges with an absorbable straight pin. Dermal wound approximation can be accomplished in significantly less time than dermal suture closure. PMID- 11495108 TI - Long-term follow-up study of bioactive bone cement for repairing a segmental defect in a canine femur. AB - We report on a 7-year long-term follow-up study of a bioactive bone cement (BA cement) that was used to repair a segmental defect in a canine femur. Bilateral femoral segmental defects were repaired with metallic implants that were fixed to the femur using two kinds of bone cement. The BA cement used in this study consists of an apatite- and wollastonite-containing glass ceramic (AW-GC) with a bis-phenol-alpha-glycidyl methacrylate (bis-GMA)-based resin. The bone-cement interface was examined histologically. Previous short-term studies have shown that using BA cement for segmental replacement of the canine femur produced excellent biomechanical and histological results. The BA cement maintained the fixation of a metallic implant to the femur very well. In contrast, the PMMA cement did not maintain alignment under long-term weight-bearing conditions. The results of histological examinations showed direct bonding between the BA cement and bone, while an intervening soft tissue layer was observed at the bone-cement interface with the PMMA cement. The BA cement bonded to the bone through a Ca-P rich reactive layer, which was twice as thick after 7 years than it was at 26 weeks. No adverse effects of BA cement were observed during the 7-year observation period. PMID- 11495109 TI - Why is p = .90 better than p = .70? Preference for definitive predictions by lay consumers of probability judgements. AB - What do people regard as an informative and valuable probability statement? This article reports four experiments that show participants to have a clear preference for more extreme and higher probabilities over less extreme and lower ones. This pattern emerged in Experiment 1, in which no context was provided, and was further explored in Experiment 2 within a positive and a negative context. The findings were further confirmed in Experiment 3, which employed a Bayesian framework with revisions of opinions. Finally, Experiment 4 showed how preference for high probabilities can lead people to prefer an overconfident to a more well calibrated (accurate) forecaster. The results are interpreted as manifestations of a search for definitive predictions principle, which asserts that high probabilities are preferred to medium ones and often favored over the corresponding complementary low probabilities on the basis of their capacity to predict the occurrence of single outcomes. PMID- 11495110 TI - Environmental context-dependent memory: a review and meta-analysis. AB - To address questions about human memory's dependence on the coincidental environmental contexts in which events occur, we review studies of incidental environmental context-dependent memory in humans and report a meta-analysis. Our theoretical approach to the issue stems from Glenberg's (1997) contention that introspective thought (e.g., remembering, conceptualizing) requires cognitive resources normally used to represent the immediate environment. We propose that if tasks encourage processing of noncontextual information (i.e., introspective thought) at input and/or at test, then both learning and memory will be less dependent on the ambient environmental contexts in which those activities occur. The meta-analysis showed that across all studies, environmental context effects were reliable, and furthermore, that the use of noncontextual cues during learning (overshadowing) and at test (outshining), as well as mental reinstatement of appropriate context cues at test, all reduce the effect of environmental manipulations. We conclude that environmental context-dependent memory effects are less likely to occur under conditions in which the immediate environment is likely to be suppressed. PMID- 11495111 TI - How the brain encodes the order of letters in a printed word: the SERIOL model and selective literature review. AB - This paper describes a novel theoretical framework of how the position of a letter within a string is encoded, the SERIOL model (sequential encoding regulated by inputs to oscillations within letter units). Letter order is represented by a temporal activation pattern across letter units, as is consistent with current theories of information coding based on the precise timing of neural spikes. The framework specifies how this pattern is invoked via an activation gradient that interacts with subthreshold oscillations and how it is decoded via contextual units that activate word units. Using mathematical modeling, this theoretical framework is shown to account for the experimental data from a wide variety of string-processing studies, including hemispheric asymmetries, the optimal viewing position, and positional priming effects. PMID- 11495113 TI - Representation of serial order in humans: a comparison to the findings with monkeys (Cebus apella). AB - In a number of studies, serially organized behavior in humans has been examined using a procedure developed for use with pigeons and monkeys. There have been few direct comparisons, however, between the data collected with humans and that collected with nonhumans, and none with respect to the interesting latency effects noted with nonhumans. The purpose of this experiment was to make this comparison. Human subjects were trained to respond to five simultaneously presented stimuli (A, B, C, D, and E) in a specific order (A-->B-->C-->D-->E) and were then tested with all 10 pairwise combinations of the five stimuli, followed by all 10 triplet combinations of the five stimuli. Mirroring the findings with monkeys (Cebus apella), humans showed a first-item effect, a missing-item effect, and a symbolic-distance effect. These results suggest that during the course of learning the five-item serial-order task humans form an internal representation of the series and access that representation to guide their behavior. PMID- 11495112 TI - Richer color experience in observers with multiple photopigment opsin genes. AB - Traditional color vision theory posits that three types of retinal photopigments transduce light into a trivariate neural color code, thereby explaining color matching behaviors. This principle of trichromacy is in need of reexamination in view of molecular genetics results suggesting that a substantial percentage of women possess more than three classes of retinal photopigments. At issue is the question of whether four-photopigment retinas necessarily yield trichromatic color perception. In the present paper, we review results and theory underlying the accepted photoreceptor-based model of trichromacy. A review of the psychological literature shows that gender-linked differences in color perception warrant further investigation of retinal photopigment classes and color perception relations. We use genetic analyses to examine an important position in the gene sequence, and we empirically assess and compare the color perception of individuals possessing more than three retinal photopigment genes with those possessing fewer retinal photopigment genes. Women with four-photopigment genotypes are found to perceive significantly more chromatic appearances in comparison with either male or female trichromat controls. We provide a rationale for this previously undetected finding and discuss implications for theories of color perception and gender differences in color behavior. PMID- 11495114 TI - Counting models of temporal discrimination. AB - A three-category task was employed to test counting models for temporal discrimination. Unlike former approaches, the present one is not based on Weber functions. Specifically, the proposed test does not require the implicit but, nevertheless, debatable assumption that the pulse rate of the internal clock is constant for different durations of the standard interval. Furthermore, the present approach does not necessitate specific distributional assumptions about the interpulse times. An experiment was conducted to evaluate the predictions of this generalized counting model. The results are consistent with predictions of the generalized counting model. A further analysis suggests that the pulse rate decreases as the duration of the standard interval increases. PMID- 11495115 TI - Stationary phantoms and grating induction with oblique inducing gratings: implications for different mechanisms underlying the two phenomena. AB - The visibility of stationary visual phantoms and the grating induction (GI) effect were concurrently analyzed with both black and gray inspection areas (IA) using the same subjects with counterbalanced orders of measurements. Oblique inducing gratings were employed in order to compare the visibility of obliquely aligned and vertically misaligned appearances between the two phenomena. Aligned and misaligned phantom responses with a black IA were similar, whereas overall phantom visibility was severely suppressed when the IA was gray. In contrast, misaligned GI dominated with a gray IA, whereas aligned and misaligned GI responses were similar with a black IA. Phantoms appear to be related to visual mechanisms' selectively utilizing relative luminance information between the inducing grating and IA in a manner consistent with more global figural characteristics of the display (e.g., modal and amodal completion). On the other hand, GI may be predominantly due to locally operating brightness/contrast mechanisms. PMID- 11495116 TI - Using magnitude estimation to investigate the perceptual components of signal detection theory. AB - Several experiments suggest that the relation between detection, signal energy, and perceived tone intensity is very different from the relation between detection, signal energy, and perceived tone duration. We propose a new task, the magnitude estimation and detection (MED) task, that allows one to assess the relation between the psychological dimensions of a stimulus and detection. In Experiment 1, we used the MED task to assess the relation between perceived tone intensity and detection in a yes/no task. The data show a strong relation between the two. In Experiment 2, we used the MED task to assess the relation between perceived tone duration and detection in a yes/no task. The data show a relatively weak relation between the two. Our data suggest that tone intensity is a less perceptually noisy dimension than tone duration. We present the advantages and disadvantages of the MED task with the hope that this task can aid researchers in better understanding the perceptual and decisional mechanisms underlying various cognitive processes. PMID- 11495117 TI - Activation and inhibition of stimulus features in conjunction search. AB - Two visual search experiments were conducted to examine how activation of target relevant features and inhibition of target-irrelevant features are involved in conjunction search. The hypothesis was that if an excitatory mechanism is involved, it should be revealed as facilitation when a target and distractors are repeated in two successive displays. If an inhibitory mechanism is involved, suppression should be obtained when distractor features from one display determine the target in the following display. The results of Experiment 1 showed that facilitation was observed consistently across two set sizes (5 and 9), whereas suppression was obtained only with larger set size (9). This pattern of results was replicated and extended in Experiment 2 with different set size. It seems that both excitatory and inhibitory mechanisms are involved in conjunction search. The excitatory mechanism seems to play a role in conjunction search regardless of search difficulty, whereas the inhibitory mechanism might play a role only when set size is larger. PMID- 11495118 TI - Eliminating flanker effects and negative priming in the flankers task: evidence for early selection. AB - Reports of negative priming in the absence of flanker effects (Fox, 1995) provide support for the notion that unattended stimuli are identified. I evaluated the hypothesis that such results are the outcome of attentional leakage to the flanker location. In Experiment 1, I assessed flanker effects and negative priming as a function of target-flanker proximity (.9 degrees and 2.7 degrees for near and far flankers, respectively) and of attention cuing to the target location (precued vs. uncued) on the prime trials. I report larger flanker effects in uncued than in precued conditions, and larger effects for near than for far flankers. More critically, when attention was precued, both flanker effects and negative priming vanished for far flankers. In Experiment 2, I show that the latter result was not linked to prime-probe contextual similarity (Neill, 1997). These results demonstrate that selective target processing is possible when attention is optimally focused to the target location. PMID- 11495119 TI - The presence of a nonresponding effector increases inhibition of return. AB - Inhibition of return (IOR) refers to the performance disadvantage for targets presented at an exogenously cued location, relative to an uncued location, at relatively long cue-target onset asynchronies. In this experiment, we investigated the influence on IOR of a nonresponding effector (i.e., the index finger of the nonresponding hand) placed on a response key in a simple-RT task. With peripheral cues and targets, IOR and spatial stimulus-response compatibility effects were larger when the nonresponding hand was placed on a response key. IOR -the slowed responding to go signals at the cued location--was accompanied by a lower false alarm rate when no-go signals were presented there. These findings provide direct evidence for a motoric component to IOR wherein some portion of the inhibition is observed as a criterion shift against responding to the cued location. PMID- 11495120 TI - Stroop interference is affected in inhibition of return. AB - In previous research, we have shown that the processing of targets that are presented to locations subject to inhibition of return (IOR) is affected by an inhibitory tagging mechanism. This mechanism acts by disconnecting activated representations of stimuli at inhibited locations from their associated responses. In two experiments, we assessed whether this inhibitory tagging mechanism of visual attention is also applied to task-irrelevant but prepotent dimensions of target stimuli, such as words in the Stroop task. To test this hypothesis, we examined the Stroop effect in an IOR procedure. The results showed that (1) IOR can be found in a color discrimination task, (2) the Stroop interference was reduced (Experiment 1) or eliminated (Experiment 2) when stimuli appeared at cued locations, as compared with cases in which they were presented at uncued locations, and (3) the effect of inhibitory tagging was limited to the shortest stimulus onset asynchrony value, replicating previous findings. These results agree with the idea that inhibitory tagging, occurring in IOR, affects the efficiency with which color words compete for responses in Stroop-like situations. PMID- 11495121 TI - The myth of ballistic processing: evidence from Stroop's paradigm. AB - The Stroop effect is widely considered to be compelling evidence that skilled readers cannot prevent themselves from reading the irrelevant word or even delay such processing. In contrast, several reports indicate that the Stroop effect can be eliminated by various simple manipulations. These reports have been criticized on several grounds, among them that the baseline condition is suspect. These criticisms are addressed by showing that when (1) a neutral baseline is replaced by congruent trials, (2) single letter cuing and coloring manipulations are combined, (3) attentional window conditions are blocked, and (4) the congruent/incongruent trial ratio is 20/80, the Stroop effect is eliminated. A second major finding is that despite no Stroop effect, negative priming is observed, consistent with the hypothesis that a distinct but delayed perceptual act processes the word. The default set may be to process to the highest level (semantics), but these reading processes are (contextually) controlled rather than ballistic. PMID- 11495122 TI - The cocktail party phenomenon revisited: the importance of working memory capacity. AB - Wood and Cowan (1995) replicated and extended Moray's (1959) investigation of the cocktail party phenomenon, which refers to a situation in which one can attend to only part of a noisy environment, yet highly pertinent stimuli such as one's own name can suddenly capture attention. Both of these previous investigations have shown that approximately 33% of subjects report hearing their own name in an unattended, irrelevant message. Here we show that subjects who detect their name in the irrelevant message have relatively low working-memory capacities, suggesting that they have difficulty blocking out, or inhibiting, distracting information. PMID- 11495123 TI - The dual-task SRT procedure: fine-tuning the timing. AB - In the standard sequential reaction time study, subjects are presented with a repeating sequence of targets to which they must respond as rapidly as possible. With practice reaction times decrease, suggesting a learned ability to exploit the repeating patterns in the display. In a common variation, a second task is interposed, typically a tone-counting task in which subjects must keep track of the frequency with which particular tones occur. In the canonical experiment, these tones appear at varying times in the interval between the subjects' response to a target and the next target. Data are presented that show that this latter variable (the RSOA, response-secondary stimulus onset asynchrony) actually plays an important but previously hidden role in these experiments. A model based on an extension of the notion of the psychological refractory period is introduced to explain these findings. PMID- 11495124 TI - Can sequence learning be implicit? New evidence with the process dissociation procedure. AB - Can we learn without awareness? Although this issue has been extensively explored through studies of implicit learning, there is currently no agreement about the extent to which knowledge can be acquired and projected onto performance in an unconscious way. The controversy, like that surrounding implicit memory, seems to be at least in part attributable to unquestioned acceptance of the unrealistic assumption that tasks are process-pure--that is, that a given task exclusively involves either implicit or explicit knowledge. Methods such as the process dissociation procedure (PDP, Jacoby, 1991) have been developed to overcome the conceptual limitations of the process purity assumption but have seldom been used in the context of implicit learning research. In this paper, we show how the PDP can be applied to a free generation task so as to disentangle explicit and implicit sequence learning. Our results indicate that subjects who are denied preparation to the next stimulus nevertheless exhibit knowledge of the sequence through their reaction time performance despite remaining unable (1) to project this knowledge in a recognition task and (2) to refrain from expressing their knowledge when specifically instructed to do so. These findings provide strong evidence that sequence learning can be unconscious. PMID- 11495125 TI - Can orthographic rimes facilitate naming? AB - Although the orthographic rime (body) is thought to play an important role in reading English, previous priming experiments found little or no evidence for facilitatory body-priming effects in the naming task. That is, hose primes NOSE no better than does a completely unrelated prime. In the present study, the hypothesis that facilitatory body-priming effects are typically masked by strong inhibitory onset effects was investigated. It was shown that when the onset of a prime was removed, facilitatory body priming could be obtained with stimuli that previously had produced no evidence of facilitation. The present study thus reconciles conflicting patterns concerning facilitation versus inhibition in body priming. PMID- 11495126 TI - Language differences in verbal short-term memory do not exclusively originate in the process of subvocal rehearsal. AB - Language differences in verbal short-term memory were investigated in two experiments. In Experiment 1, bilinguals with high competence in English and French and monolingual English adults with extremely limited knowledge of French were assessed on their serial recall of words and nonwords in both languages. In all cases recall accuracy was superior in the language with which individuals were most familiar, a first-language advantage that remained when variation due to differential rates of articulation in the two languages was taken into account. In Experiment 2, bilinguals recalled lists of English and French words with and without concurrent articulatory suppression. First-language superiority persisted under suppression, suggesting that the language differences in recall accuracy were not attributable to slower rates of subvocal rehearsal in the less familiar language. The findings indicate that language-specific differences in verbal short-term memory do not exclusively originate in the subvocal rehearsal process. It is suggested that one source of language-specific variation might relate to the use of long-term knowledge to support short-term memory performance. PMID- 11495127 TI - Social contagion of memory. AB - We report a new paradigm for studying false memories implanted by social influence, a process we call the social contagion of memory. A subject and confederate together saw six common household scenes (e.g., a kitchen) containing many objects, for either 15 or 60 sec. During a collaborative recall test, the 2 subjects each recalled six items from the scenes, but the confederate occasionally made mistakes by reporting items not from the scene. Some intrusions were highly consistent with the scene schema (e.g., a toaster) while others were less so (e.g., oven mitts). After a brief delay, the individual subject tried to recall as many items as possible from the six scenes. Recall of the erroneous items suggested by the confederate was greater than in a control condition (with no suggestion). Further, this social contagion effect was greater when the scenes were presented for less time (15 sec) and when the intruded item was more schema consistent (e.g., the toaster). As with other forms of social influence, false memories are contagious; one person's memory can be infected by another person's errors. PMID- 11495128 TI - Goal coordination in narrative comprehension. AB - In this paper, we report two experiments in which the consequence of maintaining multiple, unrelated character goals during comprehension were studied. This is in contrast to previous work that has focused on a single goal, or multiple related goals. The results showed that the presence of active subsequent goals can diminish the availability of earlier mentioned goals, even in the absence of semantic and other relations. In contrast, the status of earlier mentioned goals does not influence the availability of subsequent goals. These results support an explanation-based view of comprehension. PMID- 11495129 TI - Categorization of novel stimuli in well-known natural concepts: a case study. AB - In this study, we investigated to what extent exemplar-based and prototype predictors can be applied to predicting categorization in natural language concepts. Participants categorized novel tropical foods into two well-known natural language concepts: fruits and vegetables. The results indicate that both the prototype predictors and the exemplar predictors contribute significantly in accounting for the categorization choices but that the contribution of the prototype predictor comes from just a limited number of features. PMID- 11495130 TI - Surgical crown lengthening: a 12-month clinical wound healing study. AB - BACKGROUND: Surgical crown lengthening has been proposed as a means of facilitating restorative procedures and preventing periodontal injuries in teeth with structurally inadequate clinical crowns or exposing tooth structure in the presence of deep, subgingival pathologies which may hamper the access for proper restorative measures. The few clinical studies in the current literature on postsurgical soft tissue modifications after crown lengthening procedures report conflicting results. The present study was designed to assess the alterations of the marginal periodontal tissues as an immediate outcome of surgical crown lengthening and over a 12-month healing period. METHODS: The patient sample included 30 patients (84 teeth) who presented with various conditions hampering proper restorative measures in one or more teeth and, therefore, requiring surgical exposure of tooth substance. After initial supportive therapy, the patients were recalled for a baseline examination, and the following parameters were evaluated at interproximal and buccal/lingual sites of each experimental tooth: plaque index, gingival index, position of the gingival margin, probing depth, and attachment level. After baseline examination, the patients underwent apically positioned flap surgery with osseous and connective tissue attachment resection. During surgery, the amount of resection and the achieved lengthening of the clinical crown were evaluated. The patients were enrolled in a maintenance program including professional tooth cleaning every 2 to 4 weeks. The patients were reexamined 1, 3, 6, 9, and 12 months postoperatively. RESULTS: 1) Immediately after surgery, a significantly (P < 0.001) increased clinical crown length of 3.7 +/- 0.8 mm (mean) at interproximal and 4.1 +/- 0.9 mm (mean) at buccal/lingual sites was achieved; 2) healing resulted in a statistically significant coronal displacement of the gingival margin of 3.2 +/- 0.8 mm at interproximal (P < 0.001) and 2.9 +/- 0.6 mm at buccal/lingual (P < 0.002) sites; and 3) as a consequence of this postsurgical soft tissue regrowth, the amount of the available tooth structure immediately after surgery decreased to 0.5 +/- 0.6 mm at interproximal sites (P < 0.0015) and to 1.2 +/- 0.7 mm at buccal/lingual sites (P < 0.001) at the 12-month examination. CONCLUSIONS: The results of the present clinical investigation demonstrated that during a 1-year period of healing following surgical crown lengthening, the marginal periodontal tissue showed a tendency to grow in a coronal direction from the level defined at surgery. This pattern of coronal displacement of the gingival margin was more pronounced (P < 0.001) in patients with "thick" tissue biotype and also appeared to be influenced by individual variations in the healing response (P < 0.001) not related to age or gender. PMID- 11495131 TI - The expression of plasminogen activator system in a rat model of periodontal wound healing. AB - BACKGROUND: The plasminogen activator system has been proposed to play a role in proteolytic degradation of extracellular matrices in tissue remodeling, including wound healing. The aim of this study was to elucidate the presence of components of the plasminogen activator system during different stages of periodontal wound healing. METHODS: Periodontal wounds were created around the molars of adult rats and healing was followed for 28 days. Immunohistochemical analyses of the healing tissues and an analysis of the periodontal wound healing fluid by ELISA were carried out for the detection of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA), and 2 plasminogen activator inhibitors (PAI-1 and PAI-2). RESULTS: During the early stages (days 1 to 3) of periodontal wound healing, PAI-1 and PAI-2 were found to be closely associated with the deposition of a fibrin clot in the gingival sulcus. These components were strongly associated with the infiltrating inflammatory cells around the fibrin clot. During days 3 to 7, u-PA, PAI-1, and PAI-2 were associated with cells (particularly monocytes/macrophages, fibroblasts, and endothelial cells) in the newly formed granulation tissue. During days 7 to 14, a new attachment apparatus was formed during which PAI-1, PAI-2, and u-PA were localized in both periodontal ligament fibroblasts (PDL) and epithelial cells at sites where these cells were attaching to the root surface. In the periodontal wound healing fluid, the concentration for t-PA increased and peaked during the first week. PAI-2 had a similar expression to t-PA, but at a lower level over the entire wound-healing period. CONCLUSIONS: These findings indicate that the plasminogen activator system is involved in the entire process of periodontal wound healing, in particular with the formation of fibrin matrix on the root surface and its replacement by granulation tissue, as well as the subsequent formation of the attachment of soft tissue to the root surface during the later stages of wound repair. PMID- 11495132 TI - Orthodontic tooth movement enhances bone healing of surgical bony defects in rats. AB - BACKGROUND: The question of whether the repair of an alveolar bony defect can be enhanced by orthodontic tooth movement was addressed. METHODS: Alveolar bone defects were created in 52 Wistar male rats anterior to both maxillary first molars. After 1 week of healing, orthodontic protraction was applied for 2 weeks on the right side, resulting in mesial tipping and displacement movement. Subsequently, a retention appliance was inserted for 1 week. The left side served as the untreated (control) group. Vital bone staining (procion brilliant red H-8) was administered before and after orthodontic traction. Histomorphometric analysis was performed on 62 hemimaxillae using UV confocal microscopy and an imaging program. The total area of the bony defect was divided into 4 equal quadrants, and the area of bony apposition in each quadrant was measured. RESULTS: The total area of bony apposition was 6.5-fold larger in the treated (26.41 x 10(4) +/- 28.92 x 10(4) microm2) than in the control group (4.07 x 10(4) +/- 2.82 x 10(4) microm2), approaching statistical significance (P = 0.065). The treated occlusal quadrants demonstrated highly significant (P= 0.010), greater bone apposition compared to the control group (13.8-fold) and to the treated apical quadrants (P= 0.04, 5-fold). CONCLUSIONS: This study confirms that orthodontic tooth movement is a stimulating factor of bone apposition. A conversion in the repair pattern of the bony defect from apicoocclusal in the control group (no tooth movement) to occlusoapical in the treated group (with tooth movement) further supports the linkage between tooth movement and enhanced bone deposition. Clinical implication suggests incorporation of orthodontic tooth movement in regenerative therapy. PMID- 11495133 TI - Interleukin-1beta regulation of adhesion molecules on human gingival and periodontal ligament fibroblasts. AB - BACKGROUND: Adhesion molecules have been implicated in the pathogenesis of rheumatoid arthritis and may also play a role in the pathogenesis of periodontal disease by promoting the recruitment and retention of leukocytes in gingival tissue. METHODS: The aim of the present study was to evaluate the capacity of interleukin-1beta (IL-1beta) to regulate adhesion molecule expression on clinically healthy human gingival (HGF) and periodontal ligament (PDL) fibroblasts. The HGF (n = 6) and PDL (n = 3) fibroblasts were treated with 1.0 ng/ml of IL-1beta for 24 hours and then incubated with primary intercellular adhesion molecule-1 (ICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1) antibodies followed by FITC-conjugated secondary antibodies. The expression of ICAM-1 and VCAM-1 was measured by immunofluorescence flow cytometry. RESULTS: The levels of ICAM-1 expression in IL-1beta treated HGF and PDL fibroblasts were statistically significant (P < or = 0.05) compared to normal untreated controls using log-transformed data and 3-way analysis of variance. Both cells expressed VCAM-1 after IL-1beta treatment, but the levels were not statistically different from controls. CONCLUSIONS: This study demonstrated that IL-1beta upregulated ICAM-1 expression in both HGF and PDL fibroblasts. Even though the level of VCAM 1 was not statistically different from both HGF and PDL fibroblasts treated with IL-1beta compared to controls, both cells do express the VCAM-1 molecules. These results suggest that ICAM-1 and VCAM-1 might be involved in the pathogenesis of periodontal disease. PMID- 11495134 TI - Effects of furcation involvements on periodontal status and healing in adjacent proximal sites. AB - BACKGROUND: In a recent study it was found that following non-surgical treatment, probing depth (PD) reduction in proximal sites is significantly less in the presence of a vertical destruction in the adjacent site of the neighboring tooth. The present investigation was undertaken to study whether the presence of a proximal furcation involvement also influences the periodontal conditions in the adjacent site of the neighboring tooth. METHODS: The investigation was conducted as a retrospective study on a consecutive referral population based on full-mouth oral radiographic examinations and PD and plaque score registrations. The statistical analyses were performed on a final sample of 136 patients, with 153 (periodontal status) and 123 (periodontal healing) second maxillary premolars with an adjacent first maxillary molar. RESULTS: Baseline pockets were significantly deeper, relative radiographic attachment levels significantly reduced, and periodontal PD significantly less reduced after non-surgical treatment in the distal sites of second maxillary premolars adjacent to first maxillary molars with a mesial furcation involvement of degree > or = 2, compared to distal sites adjacent to first maxillary molars with a mesial furcation involvement of degree < or = 1. CONCLUSIONS: Periodontal status and healing after non-surgical treatment in proximal sites are negatively influenced by the presence of a deep furcation involvement in the adjacent site in the same proximal space. The presence of a deep proximal furcation involvement should consequently be considered a risk factor for the adjacent site of the neighboring tooth. PMID- 11495135 TI - Molecular analysis of Peptostreptococcus micros isolates from patients with periodontitis. AB - BACKGROUND: Recent studies provide strong evidence implicating Peptostreptococcus micros in the pathogenesis of various oral infections, including oropharyngeal abscesses and periodontal disease. To date, very little is known regarding the role of P. micros in periodontal disease. Therefore, a genetic analysis was initiated to differentiate among strains of P. micros infecting periodontal patients. METHODS: Sixty DNA samples of P. micros isolated from 15 patients with periodontal disease were evaluated. Arbitrarily primed polymerase chain reactions (AP-PCR) were performed using primer 3 (AGTCAGCCAC) and primer 13 (CAGCACCCAC). The PCR products were analyzed by gel electrophoresis. RESULTS: The primers produced several unique patterns among the strains tested. Primer 3 resulted in 30 different patterns, whereas primer 13 resulted in 31 different patterns, which were distinct from those seen with primer 3. In 8 of 15 patients, the PCR profile was identical for all isolates cultured from that patient, indicating a clonal infection. In 4 of 15 patients, 2 different genotypes were identified. In the remaining 3 patients, all isolates cultured from these patients exhibited a unique genotype. CONCLUSIONS: While P. micros appears to be heterogeneous throughout a population of periodontal patients, each patient is, for the most part, infected with a limited number of genotypes. These results demonstrate the genetic diversity of P. micros and the usefulness of AP-PCR for future epidemiological studies in understanding the role P. micros plays in periodontal disease pathogenesis. PMID- 11495136 TI - Interleukin-1beta and thiobarbituric acid reactive substance (TBARS) levels after phase I periodontal therapy in patients with chronic periodontitis. AB - BACKGROUND: Interleukin-1beta (IL-1beta), a potent stimulator of bone resorption, has been implicated in the pathogenesis of periodontal tissue destruction. There is also a clearly defined and substantial role for free radicals or reactive oxygen species in periodontal destruction. The thiobarbituric acid reactive substances (TBARS) is a commonly applied test to measure free radical activity. The aims of this study were to investigate the amount of crevicular IL-1beta, tissue TBARS levels, and the clinical status of patients with advanced chronic periodontitis and the effect of phase I periodontal therapy on these clinical parameters and measurements. METHODS: Twenty-five chronic periodontitis and 25 healthy control (C) patients were selected for the study. Plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) were recorded from each sampling area. Gingival crevicular fluid (GCF) sampling and clinical index scores were recorded at the initial examination (IE) and 6 weeks after phase I periodontal therapy (APT). Assays for GCF IL-1beta were carried out by enzyme-linked immunosorbent assay (ELISA). Gingival tissue samples were obtained from sites requiring periodontal flap surgery due to unresolved pockets to determine the tissue TBARS levels. The paired-samples t test was used to compare the IL-1beta levels and clinical parameters between IE and APT. The independent-samples t test was used to determine the significance of all parameters between IE and C, and between APT and C. The correlation among the IL 1beta levels, clinical parameters, and tissue TBARS levels was analyzed using the Pearson correlation. RESULTS: The concentration of IL-1beta levels was not statistically different among IE, APT, and C groups, but the total amount of IL 1beta levels was statistically different among the 3 groups. While the levels of IL-1beta and the clinical parameters were reduced following phase I periodontal treatment, pretreatment IL-1beta, post-treatment IL-1beta, and TBARS levels were statistically higher in IE and APT groups than C specimens. Tissue TBARS levels in the APT group were statistically greater than controls. No correlations were noted between tissue TBARS levels and clinical parameters in the APT group. A positive statistical correlation was detected between the total IL-1beta and TBARS levels in the APT group. CONCLUSION: These data suggest that the levels of crevicular IL-1beta and gingival tissue TBARS are closely associated with periodontal status. This relationship may be valuable in treating and monitoring periodontal disease progression. PMID- 11495137 TI - Cyclosporin-induced gingival overgrowth at the newly formed edentulous ridge in rats: a morphological and histometric evaluation. AB - BACKGROUND: Since cyclosporin A (CsA)-induced overgrowth seldom occurs at sites distant from teeth, the periodontal ligament has been considered significant. The aim of this study was to examine overgrowth occurrence at the edentulous ridge- the sites without the ligament--after CsA therapy in rats. METHODS: After extracting all right maxillary molars, 16 Sprague-Dawley rats underwent a 2-week healing period. The animals were separated into CsA and control groups. CsA rats received 15 mg/kg of CsA by gastric feeding for 4 weeks, while the control group received only mineral oil. At the end of study, all animals were sacrificed and stone models were immediately obtained by rubber-based impressions. The edentulous ridge morphology, including the bucco-lingual width and the vertical height, was measured on the models. For histometry, 10 sections were selected from the edentulous ridge of each animal after undecalcified tissue preparation. The soft tissue areas of the edentulous ridge and the trabecular bone morphology of the dental alveolus were measured. RESULTS: CsA therapy produced a significant increase of the ridge width and height, measured from the stone models, when compared to the control group. Under histometry, CsA resulted in a significant increase of the epithelium, connective tissue, and total soft tissue areas. The measured trabecular bone volume was affected by both examining factors: the drug therapy and the location of the dental alveolus. CsA therapy produced a significant loss of bone volume but a significant increase of the bone-specific surface area. Although the mean osteoid volume was similar between CsA and control groups, a significant decrease of the fractional formation surface in the CsA group was revealed. CONCLUSIONS: An enlarged edentulous ridge and an altered dental alveolar bone morphology were observed in CsA-treated animals at the end of the study; therefore, we suggest that CsA may induce not only a soft tissue overgrowth but also an alveolar bone alteration at the edentulous ridge. The hypothesis that tooth or periodontal ligament is an essential component for the overgrowth development is questioned. PMID- 11495138 TI - Intrapocket anesthesia for scaling and root planing: results of a double-blind multicenter trial using lidocaine prilocaine dental gel. AB - BACKGROUND: The efficacy of a novel anesthetic gel (lidocaine 25 mg/g plus prilocaine 25 mg/g in thermosetting agents) for non-invasive periodontal pocket anesthesia was evaluated. METHODS: One hundred twenty-two (122) patients in 8 centers with moderate to severe periodontitis requiring scaling and root planing (SRP) were enrolled in this multicenter, randomized, double-blind, controlled clinical trial. The active dental gel or a matching placebo gel was applied into the periodontal pocket using a blunt applicator. Following a waiting period of 30 seconds to 2 minutes, scaling and root planing were performed. If the patient had any discomfort, a second application of the gel was applied. If the patient continued to experience discomfort, conventional anesthesia was offered. After all teeth in the test quadrant had received SRP, the overall pain was assessed by the patient using a 100 mm horizontal, ungraded visual analog scale in which the left side was marked "no pain" and the right side marked "worst pain imaginable." Patients also assessed pain by using a 5-point verbal rating scale, from "no pain" to "very severe pain." RESULTS: The visual analog scale showed significant reductions in reported pain, favoring the active gel over the placebo (mean reduction, 8 mm; P <0.0005). The verbal rating scale revealed that 90% of patients treated with active gel reported no pain or mild pain compared to 64% of placebo-treated patients (P<0.001). CONCLUSIONS: Intrapocket administration of lidocaine 25 mg/g plus prilocaine 25 mg/g and thermosetting agents may be effective for pain control for scaling and root planing and may offer an alternative to infiltration anesthesia. PMID- 11495139 TI - Evaluation of the dentogingival area during early plaque formation. AB - BACKGROUND: The aims of this investigation were to evaluate the pattern of supragingival plaque formation in the dentogingival area within 96 hours after abstinence from mechanical plaque control and to clinically analyze the gingival inflammatory response observed. METHODS: Six male volunteers, 20 to 23 years of age, had their upper incisors and cuspids polished. Four independent periods of no mechanical plaque control--24, 48, 72, and 96 hours--were instituted. In each period, plaque was disclosed and standardized individual photographs were taken. Impressions were taken and replicas were analyzed by scanning electron microscopy. Gingival crevicular fluid (GCF) was collected at baseline and 96 hours. The height of the gingival papilla was measured at baseline, 24, and 96 hours. RESULTS: Results showed the presence of a plaque-free zone (PFZ) along the gingival margin up to 72 hours. After 96 hours, there was a significant reduction in the presence of PFZ in the proximal thirds of the buccal surface when compared to the other experimental periods. At the end of the study, there was a significant increase in the GCF flow as well as edema of the interdental papilla, when compared to baseline values. CONCLUSIONS: The PFZ observed during the initial phase was less apparent at the 96-hour period. At this time, there was an increase in the gingival inflammatory response, represented clinically by increased GCF flow and edema. PMID- 11495140 TI - Histopathological morphometric evaluation of 2 different hydroxyapatite-bone derivatives in sinus augmentation procedures: a comparative study in humans. AB - BACKGROUND: Xenografts to augment the maxillary sinus have been used extensively. The aim of the present study was to evaluate, qualitatively and quantitatively, two different HA derivatives of natural and synthetic sources on newly formed bone in the augmented sinus. METHODS: A bilateral sinus augmentation procedure with simultaneous (16 out of 20 sites) or subsequent implant placement was performed in 10 patients. The antrum was randomly filled with a deproteinized, bovine hydroxyapatite mineral (B-HA) on one side and a non-ceramic resorbable hydroxyapatite (NC-HA) on the other. Cylindrical specimens were harvested from the augmented core at 12 months. Decalcified specimens were sectioned at a cross horizontal plane and stained with hematoxylin and eosin for histopathologic and histomorphometric examinations. Tissue area fractions of bone, marrow, and the grafted particles were calculated for each specimen from the lateral to the deep region, and changes in values were compared within each material and between them. RESULTS: New bone formation was evident. B-HA and NC-HA particles were observed in all specimens surrounded by newly formed bone in direct connection or by soft tissue marrow. Morphometrically in the B-HA sites, from the lateral to deeper area, bone area fraction increased from 29.8% to 54.2% (average 42.1%) and marrow area fraction decreased from 37.9% to 26.7% (average 33.3%). The mineral area fraction decreased from 32.3% to 19.1% (average 24.7%). All increasing/decreasing patterns were statistically significant (P < 0.001). In the NC-HA sites, from the lateral to deeper area, bone area fraction increased from 25% to 36.5% (average 32.3%) and marrow area fraction decreased from 51.6% to 41.9% (average 43.2%) (P <0.001). The mineral area fraction decreased from 29% to 21.7% (average 24.6%) (P = 0.038). Comparison between the two HA derivative groups showed a significant difference between the bone area fraction averages (P = 0.0053) and between the increasing patterns along the core depth (P = 0.0006). There was also a significant difference between the decreasing marrow patterns (P = 0.003), but not between their averages. Comparison between the mineral area fractions showed no differences. CONCLUSIONS: B-HA and NC-HA were proven to be biocompatible materials. Although the B-HA-augmented sites showed a higher percentage of bone formation at 12 months, both are suitable bone derivatives in sinus augmentation procedures and can accommodate osseointegrated implants. PMID- 11495142 TI - Clinical evaluation of 3 techniques to augment keratinized tissue without root coverage. AB - BACKGROUND: The importance of keratinized tissue is a controversial subject. However, in some situations most clinicians would agree that surgical procedures to increase the amount of keratinized tissue without root coverage are indicated. In this study, 3 surgical procedures were compared in their ability to increase the width of keratinized tissue. They are: the epithelized autogenous masticatory mucosa graft (free gingival graft), autogenous predominately connective tissue graft (connective tissue graft), and acellular dermal matrix. METHODS: Forty-five patients referred for treatment of areas with inadequate keratinized tissue were randomly assigned into 1 of 3 groups of 15 each. Each group was treated with 1 of the 3 surgical procedures to increase the width of keratinized tissue. The width of keratinized tissue pre- and postsurgery was evaluated. RESULTS: All 3 groups started with a similar width of keratinized tissue. All of the surgical procedures resulted in a statistically significant increase in the width of keratinized tissue: free gingival graft, 4.1 mm; connective tissue graft, 3.6 mm; and acellular dermal matrix, 4.1 mm. CONCLUSION: A statistically significant increase in the amount of keratinized tissue was obtained with all 3 surgical procedures evaluated. PMID- 11495143 TI - Excision and repair of the peripheral ossifying fibroma: a report of 3 cases. AB - BACKGROUND: The peripheral ossifying fibroma (POF), one of the most common gingival lesions, has a recurrence rate of nearly 20%. To minimize the reappearance of this lesion, it must be completely excised. In the maxillary anterior region, total excision of a POF can result in an unsightly gingival defect. METHODS: Three cases are presented in which a POF was excised from the gingiva facial to a maxillary central incisor. One of these lesions had previously undergone 2 cycles of conservative excision and recurrence. In all cases, the lesions were excised down to bone. Each of the resulting gingival defects was repaired by a distinct plastic surgery procedure, including a laterally positioned flap, a subepithelial connective tissue graft, and a coronally positioned flap. RESULTS: The defects resulting from the biopsies were satisfactorily repaired. The patients were followed over postsurgical intervals of 10 to 30 months. None of the lesions recurred. CONCLUSIONS: It is customary to manage POF by aggressive excisional biopsy. Several different surgical approaches may potentially be used to repair the resultant gingival defect and minimize patient esthetic concerns. PMID- 11495141 TI - Connective tissue growth factor in drug-induced gingival overgrowth. AB - BACKGROUND: Drug-induced gingival overgrowth is a known side effect of certain chemotherapeutic agents used for the treatment of systemic disorders. The pathogenesis and mechanisms responsible for this condition are not fully understood. This study assesses for the presence and localization of connective tissue growth factor (CTGF) in drug-induced gingival overgrowth tissues. CTGF immunostaining was compared with sections stained with transforming growth factor (TGF)-beta1 and CD31 antibodies in order to investigate possible pathogenic mechanisms. METHODS: Gingival overgrowth samples were obtained from patients undergoing therapy with phenytoin (n = 9), nifedipine (n = 4), cyclosporin A (n = 5), and control tissues from systemically healthy donors (n = 9). Tissue sections were subjected to peroxidase immunohistochemistry and were stained with CTGF and TGF-beta1 polyclonal primary antibodies. Possible relationships between CTGF staining and angiogenesis were also studied using an anti-CD31 antibody as a marker for endothelial cells. Staining was analyzed by computer-assisted quantitative and semiquantitative methodology at 5 defined sites in all samples based on the location of specific landmarks including epithelium and underlying connective tissues. RESULTS: Cellular and extracellular CTGF content in phenytoin gingival overgrowth tissues was significantly (P<0.05) higher compared to the other gingival overgrowth tissues and the controls. Higher CTGF staining in phenytoin gingival overgrowth tissues was accompanied by an increased abundance of fibroblasts and connective tissue fibers. No strong association of CTGF staining with TGF-beta1 or CD31 staining was found. CONCLUSIONS: The data from the present study show significantly higher CTGF staining in phenytoin-induced gingival overgrowth tissues compared to controls, cyclosporin A-, or nifedipine induced gingival overgrowth. Moreover, semiquantitative analyses of histologic samples support the concept that the phenytoin overgrowth tissues are fibrotic. These associations suggest a possible role for CTGF in promoting development of fibrotic lesions in phenytoin-induced gingival overgrowth. PMID- 11495145 TI - Erythema multiforme secondary to herpes simplex infection: a case report. AB - BACKGROUND: Erythema multiforme (EM) is a complex disease that may have cutaneous and/or mucosal involvement. The severity may range from mild to severe and potentially life threatening. The literature cites many factors including viruses, infections, and medications as causes. This report documents a patient who developed EM secondary to a herpes simplex viral (HSV) infection. METHODS: Two weeks following an eruption of herpes labialis, a 20-year-old white female patient developed acutely painful oral and labial ulcers accompanied by target skin lesions. A diagnosis of erythema multiforme (EM) was made. The patient was treated with antivirals, analgesics, and symptomatic therapy. RESULTS: Nine days after the onset of symptoms, the oral and cutaneous lesions had started to heal and the patient no longer required pain medication. CONCLUSIONS: Although the etiology of EM is still often unknown, infections with herpes simplex virus have been implicated as a possible precipitating factor. This case illustrates the association of the occurrence of EM with an HSV infection. PMID- 11495144 TI - Clinical periodontal findings and microflora profiles in children with chronic neutropenia under supervised oral hygiene. AB - BACKGROUND: This is the first known case report that used a polymerase chain reaction (PCR)-based method to help identify the oral microflora in patients with chronic neutropenia. In this study, we report clinical periodontal findings and microflora profiles of 2 children, 1 with severe congenital neutropenia (SCN, Kostmann type) and 1 with cyclic neutropenia (CN). METHODS: The SCN patient had severe gingivitis, whereas the patient with CN had mild gingivitis in the gingival margins. Monthly oral cleaning instruction and review were performed without subsequent periodontal therapy. Oral hygiene conditions remained satisfactory and visible plaque was scarce, despite the persistence of mild gingivitis. Under supervised oral hygiene, we examined the presence of periodontal pathogens from patient plaque samples. RESULTS: By a PCR-based method, Prevotella nigrescens, Bacteroides forsythus, Campylobacter rectus, and Capnocytophaga gingivalis were detected in the SCN patient and P. intermedia, C. rectus, C. gingivalis, and C. sputigena in the CN patient, suggesting the existence of periodontal pathogens. Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, and C. ochracea were not found in either patient. CONCLUSIONS: Use of 1% povidone iodine solution and local antibiotic application under supervised oral hygiene were helpful to improve gingival conditions in patients with chronic neutropenia. PMID- 11495146 TI - Interimplant papilla reconstruction: assessment of soft tissue changes and results of 12 consecutive cases. AB - BACKGROUND: Loss of soft tissue between adjacent implants is a frequent occurrence. Reconstruction of the papilla between adjacent implants is often necessary where there are esthetic demands. The purpose of this article is to describe a fixed and reproducible reference line which can be used to measure changes in the height of the interimplant papillae. A surgical technique carried out at the time of the abutment connection surgery is also described to show the changes that may occur over a 12-month period. Assessment of changes in the soft tissue dimension is shown following a modified flap technique at the time of abutment connection. METHODS: With the aid of clinical photographs taken preoperatively, a reference line can be drawn between the highest point of the gingival margins on teeth distal to the intended site of soft tissue surgery. This reference line can be reproduced on clinical photographs postoperatively. Using this reference line, the changes in soft tissue height using a modified flap technique in 12 consecutive patients are presented. Assessment of the vertical height of the interimplant papillae before and after treatment is described. RESULTS: Four patients showed an increase in the height of the papillae when reviewed 12 months following abutment connection. One patient showed no change in the height of the papillae, and 7 patients showed a loss in papillary height. There was an average loss of 0.5 mm in height of the interimplant soft tissue. CONCLUSIONS: Use of a fixed reference line to assess the outcome of treatment is necessary when evaluating vertical changes in soft tissue. This reference line is used to assess a soft tissue flap technique carried out at the time of implant exposure. PMID- 11495147 TI - Tooth mobility revisited. PMID- 11495148 TI - Guidelines: In-office use of conscious sedation in periodontics. AB - In this time of heightened awareness of periodontal diseases and the potential consequences of untreated disease, a deterrent in the delivery of periodontal care continues to be patient anxiety concerning treatment and the fear of pain. These guidelines are intended for periodontists in the in-office use of enteral, inhalation, and/or parenteral conscious sedation in the delivery of care. The definitions, educational guidelines, and policies presented in these guidelines are consistent with the most current American Dental Association (ADA) documents Guidelines for the Use of Conscious Sedation, Deep Sedation and General Anesthesia for Dentists and the Guidelines for Teaching the Comprehensive Control of Anxiety and Pain in Dentistry available from the American Dental Association, 211 E. Chicago Avenue, Chicago, IL 60611 or http://www.ada.org, and for Revisions to Anesthesia Care Standards Comprehensive Accreditation Manual for Ambulatory Care, effective January 1, 2001, Joint Commission on Accreditation of Health Care Organizations, available through http://www.jcaho.org/standard/anesamb.html. This paper replaces the former position paper entitled "Guidelines for the Use of Conscious Sedation in Periodontics." PMID- 11495149 TI - Evaluation of an Estimation of Physiologic Ability and Surgical Stress (E-PASS) scoring system to predict postoperative risk: a multicenter prospective study. AB - We previously reported generating a scoring system termed E-PASS that predicted postsurgical risk. This study was undertaken to evaluate the usefulness of this system. A consecutive series of 902 patients who underwent elective gastrointestinal operations in six national hospitals in Japan were prospectively assessed for a comprehensive risk score (CRS) of the E-PASS, which was compared with their postoperative course. The postoperative morbidity rates linearly increased as the CRS increased. The postoperative mortality rate was only 0.13%, when the CRS was below 0.5; however, it increased to 9.7% when the CRS ranged from 0.5 to <1.0, and to 26.9% when the CRS was > or =1.0. The CRS correlated significantly with the severity of postoperative complications (rs = 0.527, P < 0.0001) and the costs of hospital stay (rs = 0.810, P < 0.0001). When the CRS adjusted mortality rate at the CRS of > or =0.5 was compared among the hospitals, it was related to the hospital volume of operations, being 44.2% at the volume of <100 cases per year, 20.6% at the range of 100-199 cases, and 8.6% at the volume of > or =200 cases. These results suggest that E-PASS may be useful for predicting postsurgical risk, estimating medical expense, and comparing surgical quality. PMID- 11495150 TI - Increased plasma concentration of adrenomedullin during and after major surgery. AB - Adrenomedullin (AM), a novel vasodilatory peptide, appears to participate in the regulation of circulation. Recent studies have reported a relationship between AM and cytokine. The present study was performed to determine whether major surgery, which is accompanied by an altered cytokine production, affects the concentration of plasma AM. Eighteen patients undergoing major upper abdominal and thoracoabdominal surgery were thus studied. Arterial blood samples were obtained before anesthetic induction (PRE), 2h after a surgical incision was made (2H), at the end of surgery (END), at postoperative day 1 (1POD), and at 3POD. The concentrations of plasma AM and serum interleukin (IL)-6 were measured. The levels of plasma AM were found to be significantly higher at END (P < 0.001), 1POD (P < 0.0001), and 3POD (P < 0.05) compared with PRE. A significant correlation was found between the concentrations of serum IL-6 and plasma AM (r = 0.59, n = 18, P < 0.01). These results suggest that major surgical stress might thus increase the plasma AM concentration, and such an increase may be related to cytokine induction. PMID- 11495151 TI - The outcome of volume reduction surgery according to the underlying type of emphysema. AB - Lung volume reduction surgery (VRS) is widely performed to improve symptoms in patients with severe pulmonary emphysema; however, it remains unclear whether or not the underlying type of emphysema affects the surgical results. A total of 58 patients with advanced symptomatic emphysema underwent thoracoscopic VRS of emphysematous lung tissue. The resected lung tissue was examined microscopically, and the patients were classified into two groups according to the predominant pathological findings, as having either centrilobular-type or panacinar-type emphysema. A predominant pathological type was identified in only 34 patients (59%), 19 of whom had centrilobular-type emphysema and 15, panacinar-type emphysema. Patients with predominately panacinar emphysema had more compromised total lung capacity and residual volume than those with centrilobular emphysema. Significant improvements in forced expiratory volume in 1 s were evident 3 months after surgery in the group with centrilobular emphysema (+515 +/- 141ml) compared with that in the group with panacinar-type emphysema (+109 +/- 40ml, P = 0.03). The results of lung VRS were found to depend on the underlying type of emphysema, as the benefits of surgery for panacinar emphysema were less marked than those for centrilobular emphysema. PMID- 11495152 TI - Modified ultrafiltration may improve postoperative pulmonary function in children with a ventricular septal defect. AB - To evaluate the effectiveness of modified ultrafiltration (MUF) on ventricular septal defect (VSD) repair in children, we retrospectively examined 10 patients who underwent VSD repair with MUF at the Kinki University School of Medicine hospital between June 1998, and December 1998 (MUF group). These patients were compared with 14 patients who underwent the same procedure without MUF (control group) between January 1997 and June 1998. Systolic blood pressure and hematocrit values increased significantly during MUF. By the time of postoperative transfer to the intensive care unit, PaO2 was higher in the MUF group than in the control group (503.3 +/- 112.2mmHg vs 376.3 +/- 149.2mmHg; P = 0.0491), whereas A-aDO2 was lower in the MUF group than in the control group (171.9 +/- 109.2mmHg vs 301.1 x 150.4mmHg; P = 0.0449). These findings demonstrate that MUF had a beneficial effect on pulmonary function in children who underwent surgery to repair a VSD. PMID- 11495153 TI - Presence of serum p53 antibodies is associated with decreased in vitro chemosensitivity in patients with esophageal cancer. AB - Resistance to chemotherapy remains a serious problem inhibiting the successful treatment of advanced esophageal cancer. A number of studies have revealed that p53 genetic alteration and protein overexpression can predict chemosensitivity. Furthermore, p53 protein overexpression in cancer tissues has been found to induce serum p53 antibodies (p53-Abs). This study was conducted to examine whether analysis of serum p53 Abs could predict the chemosensitivity of esophageal cancer. Serum analysis of p53 antibodies was performed by enzyme linked immunosorbent assay in 19 patients with esophageal squamous cell carcinoma preoperatively, then surgically resected specimens were stained immunohistochemically for p53 protein expression. Tumor tissues were also analyzed for chemosensitivity by the histoculture drug response assay (HDRA) using cis-dichlorodiammineplatinum(II) (CDDP), 5-fluorouracil (5-FU), and adriamycin (ADM). Serum p53-Abs were present in 47% (9/19) of the patients and immunohistochemical analysis revealed overexpression of p53 protein in 42% (8/19) of the tumors. The presence of serum p53 antibodies was significantly correlated with p53 immunoreactivity (P = 0.005). The inhibition index of patients positive for p53-Abs was significantly lower than that of patients negative for p53-Abs (P < 0.001). This tendency was also observed in the inhibition index to 5-FU. The presence of serum p53-Abs was associated with decreased in vitro chemosensitivity to CDDP and 5-FU. Thus, the detection of serum p53-Abs is suggested to be useful for predicting chemosensitivity in patients with esophageal cancer. PMID- 11495154 TI - Preclinical study of adenoviral p53 gene therapy for esophageal cancer. AB - An alteration of the p53 gene function is a major factor in the development of esophageal cancer. Recently, p53 gene therapy has been applied for clinical studies in lung cancer and head and neck cancer. However, no preclinical studies have yet demonstrated an anticancer effect of adenoviral-mediated wild-type p53 gene therapy on esophageal cancer. We herein evaluated the effect of p53 adenoviral gene therapy on human esophageal squamous cell carcinoma to test the ability of clinical application. A normal esophageal epithelial cell line (EN53F) and two human esophageal cancer cell lines (ECGI-10 and T.Tn) with a p53 alteration were used. The transduction efficiency, p53 protein expression, p21 protein expression, the induction of apoptosis, and growth suppression were assessed by using the recombinant adenoviral vector Ad5CMV-p53. The transduction efficiency was 60%-80% at 100 plaque-forming units (PFU)/cell and 80%-100% at 300PFU/cell. A significant growth suppression following an Ad5CMV-p53 infection was observed in both cancer cell lines. A Western blot analysis confirmed the presence of both exogenous p53 protein expression and p21 protein induction. Apoptotic cell death was observed with TUNEL staining. T.Tn xenografts in nude mice transduced with Ad5CMV-p53 demonstrated significant growth suppression. These data suggest that Ad5CMV-p53 may thus be a potentially effective therapeutic agent for locally advanced esophageal cancer. PMID- 11495155 TI - Systemic macrophage migration inhibitory factor release following hepatic resection. AB - We set out to determine the responses of macrophage migration inhibitory factor (MIF) to hepatic resection and investigate its role in predicting short-term postoperative morbidity and outcome. Blood samples from 29 patients undergoing hepatic resection and eight healthy volunteers were obtained serially for 24h and assayed for serum MIF, cortisol, and tumor necrosis factor (TNF)-alpha. The MIF and cortisol levels showed a parallel increase and their peak levels were significantly correlated (r2 = 0.33, P = 0.0011). The TNF-alpha levels also increased during and after hepatic resection, but did not correlate with the MIF levels. The patients were classified into an extended hepatectomy group (n = 9); a lobectomy/segmentectomy group (n = 12); and a subsegmentectomy group (n = 8). There were no differences in the time courses of MIF (P = 0.8699), cortisol (P = 0.7485), and TNF-alpha (P = 0.3819) among the three groups. No patients developed organ dysfunction and all were discharged from the intensive care unit without any complications. Our findings demonstrate that MIF production occurs in patients undergoing hepatic resection. Surgical stress may play a more important role in MIF secretion than inflammatory stimulus by TNF-alpha in these patients. Therefore, MIF minimally affects short-term postoperative morbidity and outcome. PMID- 11495156 TI - Comparison of Bassini repair and mesh-plug repair for primary inguinal hernia: a retrospective study. AB - The purpose of this study was to compare the mesh-plug repair with the Bassini repair for the treatment of primary unilateral inguinal hernias. Patients with primary unilateral inguinal hernias who underwent a Bassini repair (n = 118) between January 1992 and May 1996 and a mesh-plug repair (n = 113) between July 1996 and April 1998 were retrospectively reviewed. We recorded information regarding the types of hernia according to Nyhus classification, operation time, complications, postoperative recovery, and recurrence after surgery. The two groups were comparable regarding age, sex, side of hernia, types of hernia, and the follow-up interval. The operation time was 55 +/- 20min for Bassini repair and 54 +/- 18min for mesh-plug repair. There was no incidence of mesh infection in the mesh-plug repair cases. The amount of diclofenac sodium (suppository) was 307 +/- 222mg in the Bassini repair group and 132 +/- 182mg in the mesh-plug repair group (P < 0.0001). The length of hospital stay was 8.2 +/- 2.0 days in the Bassini repair group and 4.3 +/- 2.7 days in the mesh-plug repair group (P < 0.01). Nine patients (7.6%) in the Bassini repair group had recurrence, compared with one patient (0.9%) in the mesh-plug repair group. The recurrence-free survival in the mesh-plug repair group was significantly longer than that in the Bassini repair group (P = 0.03). In conclusion, patients with primary unilateral inguinal hernias who undergo a mesh-plug repair recover more rapidly and have less recurrence in comparison with those who undergo a Bassini repair. PMID- 11495157 TI - Route of feeding influences the production and expression of tumor necrosis factor alpha in burned rats. AB - The effect of nutritional route on tumor necrosis factor alpha (TNF-alpha) production in burned rats was examined. Scald burns covering about 30% of the whole body surface area were inflicted on 43 male Wistar rats weighing about 200g. The animals were divided into three groups: CHOW (n = 10), total parenteral nutrition (TPN) (n = 22), and total enteral nutrition (TEN) (n = 11), continuously given a chow diet, TPN solution, or an enteral diet, respectively, for 7 days after the burn injury. The rate of detection of TNF-alpha in plasma on day 7 was significantly higher in the TPN group than in the CHOW or TEN groups. The messenger RNA (mRNA) expression of TNF-alpha was significantly increased in the spleen, lungs, liver, and ileum of the rats receiving TPN compared with the CHOW and TEN rats. On the other hand, the expression of TNF-alpha mRNA was markedly decreased in the thymus of the TPN group compared with the CHOW group. The mortality rate in the TPN group (63.6%) was higher than that in the CHOW (0.0%) or TEN (27.3%) groups on day 7 after burn injury. These data suggest that TPN increases the expression of TNF-alpha mRNA in organ tissues and systemic TNF alpha production, and reduces the survival rate of rats after thermal injury, but TEN does not. PMID- 11495158 TI - Bilirubin calculi crushing by laser irradiation at a molecular oscillating region wavelength based on infrared absorption spectrum analysis using a free-electron laser: an experimental study. AB - We investigated a new laser technique of crushing bilirubin calculi, our aim being to crush calculi in isolation using a minimally invasive procedure. Infrared absorption spectrum analysis of the bilirubin calculi was conducted, revealing maximum absorption spectrum at a wavelength of the C=O stretching vibration of ester binding that exists within the molecular structure of bilirubin calcium. As an experiment to crush calculi using the free-electron laser, we set the laser at the effective irradiation wavelength of ester binding, and conducted noncontact irradiation of the bilirubin calculi. The calculi began to slowly ablate until the irradiated site had been completely obliterated after 20s of irradiation. Moreover, absorption spectrum analysis of the irradiated site, from a comparison of absorption peak ratios, revealed that absorption peak intensities decreased over time at the absorption wavelength of ester binding. These findings suggest that irradiation of molecular oscillating region wavelengths peculiar to calculi based on infrared absorption spectrum analysis results in the gradual crushing of calculi in isolation by breaking down their molecular structure. PMID- 11495159 TI - Early microcirculatory derangement in mild and severe pancreatitis models in mice. AB - An in vivo microscopic technique was used to clarify the increase in microvascular permeability and enhanced leukocyte-endothelium interaction of pancreatic microcirculation in experimental pancreatitis of differing severity. Using bovine albumin fluorescein isothiocyanate (FITC) and carboxyfluorescein diacetate succinimidyl ester (CFDASE) as tracers, the change in permeability and the behavior of leukocytes in the acinar microcirculation were quantified during the initial 1, 2, 6, and 12h after the induction of caerulein pancreatitis in mice. Cold stress was added to produce the severe model. It was revealed that the early microcirculatory changes in the pancreas of caerulein pancreatitis included the increased permeability of endothelial lining and an accumulation of extravasated fluid in the perilobular space, which were more severe if cold stress was added. A decrease in flow velocity was also noted 2h after the onset of severe pancreatitis. Leukocyte adherence to the endothelial cells was not observed during the first 12h in either model of severity. In contrast, observation of the hepatic microcirculation revealed a significant number of adherent leukocytes 2h after the induction of severe pancreatitis. These results suggest that during the early course of acute pancreatitis, leukocyte adherence in the pancreatic microcirculation is a secondary event following the increase in pancreatic vascular permeability. PMID- 11495160 TI - A reoperation for anomalous origin of right pulmonary artery: report of a case. AB - A reoperation to upsize the conduit placed at infancy for the repair of an anomalous origin of the right pulmonary artery (AORPA) was successfully performed in an 8-year-old girl because of an elevated right ventricular pressure and a reduced right pulmonary blood flow. Although primary direct anastomosis is essential for AORPA, one should not hesitate to perform a conduit repair (interposition with a tube prosthesis) on an infant with AORPA whose right pulmonary artery is distant from the main pulmonary artery, because a reoperation can safely be performed even in cases where the conduit is relatively narrow as the patient grows. This is the first report of a reoperation, including a complete replacement of the conduit, after an initial conduit repair for AORPA. PMID- 11495161 TI - Disseminated peritoneal adenomucinosis associated with a panperitonitis-like onset: report of a case. AB - A 59-year-old man was admitted to our department due to a fever of unknown origin. Abdominal ultrasonography and computed tomography showed a large cystic mass in the lower abdomen and a massive amount of abdominal fluid. A laparotomy was performed under a diagnosis of panperitonitis. Diffuse pyogenic gelatinous ascites and a large cystic mass with a grayish wall, and a hard solid lesion in part were found. The microscopic findings of the hard solid lesion showed calcification, osteogenesis, and focal epithelial proliferation in a tiny area consisting of mucinous cells with no significant cytologic atypia. The remaining part of the cystic wall and small cystic lesions were hyalinized, fibrous, or necrotic tissue. Since a total resection of the masses was not possible, the patient received adjuvant chemotherapy with cisplatin followed by the administration of mitomycin C and 5-fluorouracil. An abdominal fistula with the excretion of pyogenic gelatinous fluid occurred, but the patient is still alive and doing well over 2 years postoperatively. The primary site of this tumor could unfortunately not be identified. PMID- 11495162 TI - Hepatobiliary cystadenoma combined with multiple liver cysts: report of a case. AB - Hepatobiliary cystadenomas are rare benign tumors with malignant potential. They are almost always solitary lesions accompanied by multilocular cysts in the liver, and are difficult to differentiate from cystadenocarcinoma, despite the diagnostic modalities available. This report describes a case of hepatobiliary cystadenoma with multiple cysts in the left hepatic lobe, diagnosed by magnetic resonance imaging in a 48-year-old woman. Abdominal computed tomography revealed only multiple cystic lesions in the left lobe, but cholangiography via a nasogastric biliary drainage tube combined with percutaneous transhepatic cholangiography showed a stenotic region with fine irregularity in the left lateral posterior segmental bile duct and left lateral anterior segmental bile duct. Hepatobiliary cystadenocarcinoma with multiple liver cysts was suspected. We performed left hepatectomy, and microscopic examination confirmed a diagnosis of hepatic cystadenoma with multiple liver cysts. There was no nuclear atypia or mitosis in the epithelium of the locus, which was constructed of simple columnar to-cuboidal epithelium with basal nuclei. The patient is well without recurrence more than 4 years after surgery. PMID- 11495163 TI - An immunohistochemical examination of basaloid squamous cell carcinoma of the esophagus: report of a case. AB - We recently encountered a patient with basaloid carcinoma of the esophagus with extensive node involvement. The patient died of hematogenous metastasis 6 months after surgery. The tumor expressed cytokeratin but did not express either Type IV collagen or laminin. Both tumor cells and metastatic lesions in the regional lymph nodes expressed p53, Bcl-2, and Ki-67 proteins, but did not express cyclin D1 proteins. PMID- 11495164 TI - Ruptured aneurysm of the visceral artery: report of two cases. AB - We present herein two cases of a ruptured aneurysm of the visceral artery. The first case involved a 74-year-old man with abdominal pain who was admitted to our hospital with a tentative diagnosis of intra-abdominal bleeding of unknown origin. Computed tomography revealed a hematoma in the greater curvature of the stomach. At surgery, a hematoma along the right gastroepiploic artery was found and totally removed. Histological examination showed a pseudo-aneurysm of unknown etiology. The second case involved a 68-year-old man with progressive anemia who presented with spontaneous intra-abdominal bleeding. A ruptured aneurysm of the accessory middle colic artery was diagnosed by superior mesenteric angiography. The ruptured aneurysm was ligated and totally resected without a colectomy. Histological examination showed a pseudoaneurysm of unknown etiology. The postoperative courses were uneventful, and both patients were doing well at the time of writing. PMID- 11495165 TI - How does cognitive therapy prevent relapse in residual depression? Evidence from a controlled trial. AB - This study examined the cognitive mediation of relapse prevention by cognitive therapy (CT) in a trial of 158 patients with residual depression. Scores based on agreement with item content of 5 questionnaires of depression-related cognition provided no evidence for cognitive mediation. A measure of the form of response to those questionnaires, the number of times patients used extreme response categories ("totally agree" and "totally disagree"), showed significant and substantial prediction of relapse, differential response to CT. and conformity to mediational criteria. CT reduced relapse through reductions in absolutist, dichotomous thinking style. CT may prevent relapse by training patients to change the way that they process depression-related material rather than by changing belief in depressive thought content. PMID- 11495166 TI - Psychopathy and community violence among civil psychiatric patients: results from the MacArthur Violence Risk Assessment Study. AB - Although psychopathy is recognized as a relatively strong risk factor for violence among inmates and mentally disordered offenders, few studies have examined the extent to which its predictive power generalizes to civil psychiatric samples. Using data on 1,136 patients from the MacArthur Violence Risk Assessment project, this study examined whether the 2 scales that underlie the Psychopathy Checklist: Screening Version (PCL:SV) measure a unique personality construct that predicts violence among civil patients. The results indicate that the PCL:SV is a relatively strong predictor of violence. The PCL:SV's predictive power is substantially reduced, but remains significant, after controlling for a host of covariates that reflect antisocial behavior and personality disorders other than psychopathy. However, the predictive power of the PCL:SV is not based on its assessment of the core traits of psychopathy, as traditionally construed. Implications for the 2-factor model that underlies the PCL measures and for risk assessment practice are discussed. PMID- 11495167 TI - Short- and long-term effectiveness of an empirically supported treatment for agoraphobia. AB - This study examined the effectiveness of individual high-density exposure (2-3 weeks, all day) for panic disorder with agoraphobia (PDAG). Participants were 416 unselected patients with a primary diagnosis of PDAG who were treated by 52 therapists in 3 outpatient clinics of the Christoph-Dornier Foundation of Clinical Psychology in Germany. Results 6 weeks after the end of therapy and at the 1-year follow-up showed highly significant reductions in anxiety symptoms, anxious cognition, agoraphobic avoidance, general symptomatology, and depressive symptoms. Results did not differ significantly between the 3 outpatient clinics and are comparable with the average effect sizes reported by meta-analytic studies of controlled efficacy research, using selected patients and specifically trained therapists. Effectiveness was not dependent on duration of disorder, number of treatment sessions, and therapist experience. The study suggests that high-density exposure can be transported from research settings to the mental health field. PMID- 11495168 TI - Bias in binge eating disorder: how representative are recruited clinic samples? AB - The aim of this study was to investigate sampling bias as it affects recruited clinic samples of binge eating disorder (BED). Demographic and clinical characteristics of a recruited clinic sample were compared with a community sample. The 2 groups met the same operational definition of BED and were assessed using the same primarily interview-based methods. Ethnicity, severity of binge eating, and social maladjustment were found to increase treatment seeking among participants with BED rather than levels of psychiatric distress or comorbidity. These findings suggest that previous studies using recruited clinic samples have not biased estimates of psychiatric comorbidity in BED. PMID- 11495169 TI - Meta-analysis of HIV risk-reduction interventions within drug abuse treatment programs. AB - A meta-analysis was conducted on studies using a treatment-comparison group design to evaluate HIV/AIDS risk-reduction interventions for clients enrolled in drug abuse treatment programs. Overall, the interventions studied were found to have a reliable positive (weighted) effect size (d = 0.31), and this was unlikely to be due to publication bias. Effect sizes for specific categories of outcome variables were 0.31 for knowledge, attitudes, and beliefs; 0.26 for sexual behavior; 0.62 for risk-reduction skills; and 0.04 for injection practices. A number of potential moderators were examined. Effect sizes were negatively correlated with the presence of predominantly ethnic minority samples and positively correlated with the number of intervention techniques used, the intensity of the intervention, intervention delivery at a later stage of drug treatment or within methadone treatment, and the presence of a number of specific intervention techniques. PMID- 11495170 TI - The relationship of caregiver coping to family outcomes during the initial year following pediatric traumatic injury. AB - This study identified coping strategies associated with caregiver outcomes following pediatric injury and examined injury type as a moderator of coping efficacy. Families of 103 children with traumatic brain injury (TBI) and 71 children with orthopedic injuries were followed prospectively during the initial year postinjury. The groups had comparable preinjury characteristics and hospitalization experiences but differed on neurological insult. In hierarchical regression analyses, acceptance was associated with lower burden and denial was associated with greater distress in both groups. Active coping resulted in higher distress following TBI but not orthopedic injuries. Conversely, the use of humor was related to diminishing distress following TBI but unrelated to distress following orthopedic injuries. Results are discussed in terms of the implications for intervention following TBI. PMID- 11495171 TI - Preventing problems with boys' noncompliance: effects of a parent training intervention for divorcing mothers. AB - This study provided a randomized, experimental test of the efficacy of a parent training intervention on coercive discipline, positive parenting practices, and child noncompliance in a sample of 238 divorcing mothers and their sons in Grades 1-3. Intervention effects were evaluated 5 times from baseline to 30 months. The intervention produced enduring benefits to coercive discipline, positive parenting, and boys' noncompliance. These benefits followed a classic prevention effect: Mothers and sons in the experimental group maintained stable outcome trajectories, whereas those in the control group deteriorated. The intervention's impact on boys' noncompliance was mediated independently by its impact on coercive discipline and positive parenting. Change in positive parenting was more strongly associated with change in noncompliance than was change in coercive discipline, although each explained unique variance in change in noncompliance. PMID- 11495172 TI - Strike while the iron is hot: can stepped-care treatments resurrect relapsing smokers? AB - The efficacies of 2 group counseling step-up treatments for smoking cessation, cognitive-behavioral/skill training therapy (CBT) and motivational interviewing/supportive (MIS) therapy, were compared with brief intervention (BI) treatment in a sample of 677 smokers. Differential efficacy of the 2 step-up treatments was also tested in smokers at low and high risk for relapse (no smoking vs. any smoking during the first postquit week. respectively). All participants received 8 weeks of nicotine patch therapy. BI consisted of 3 brief individual cessation counseling sessions; CBT and MIS participants received BI treatment and 6 group counseling sessions. Neither CBT nor MIS treatment improved long-term abstinence rates relative to BI. Limited support was found for the hypothesis that high-risk smokers would benefit more from MIS than CBT. Other hypotheses were not supported. PMID- 11495173 TI - A comparison of weight control and weight control plus body image therapy for obese men and women. AB - Participants were 65 obese men and women who were randomly assigned to either weight control or weight control plus cognitive behavioral body image therapy. Both conditions showed clinically significant improvements in body image at posttreatment and 1-year follow-up. Adding body image therapy to weight control did not result in greater psychological improvements and did not result in better maintenance of body image change when participants regained weight after treatment. Weight loss and maintenance were equivalent between groups. Adding body image therapy did not improve or detract from weight loss. Although body image therapy has been shown to be effective in obese persons, it appears that a well-rounded cognitive-behavioral weight control program is effective as well. PMID- 11495174 TI - Reducing alcohol consumption among heavily drinking women: evaluating the contributions of life-skills training and booster sessions. AB - Problem-drinking women (N = 144) without histories of severe physical dependence on alcohol received drinking-reduction training and were assigned to receive (or not receive) 2 treatment enhancements: life-skills training and booster sessions. The design resulted in 4 treatment conditions: drinking-reduction treatment (DRT) plus life-skills training, DRT plus booster sessions, DRT plus life-skills training and booster sessions, or DRT only. The interventions entailed 13 hr of DRT, 7 hr devoted to the life-skills training or to a no-life-skills training educational module, plus 8 hr of booster sessions for those receiving them. Participants evidenced significant reductions in alcohol use during the 18 months after treatment. Those with greater pretreatment drinking evidenced differential response to the experimental manipulations: The treatment enhancements (life skills and booster sessions) led to significantly improved drinking outcomes among women who were heavier drinkers at pretreatment. There were no significant effects of the treatment enhancements among lighter drinkers at pretreatment. The results provide support for use of treatment enhancements in interventions designed to moderate women problem drinkers' alcohol use. PMID- 11495175 TI - A contextual analysis of externalizing and mixed syndrome boys: when syndromal similarity obscures functional dissimilarity. AB - A contextualized approach to psychopathology that focuses on children's responses to events rather than overall behavior output was examined. Teachers rated boys in special education classrooms using a standardized syndromal instrument and one that assessed boys' social environments and their responses to them. Externalizing and mixed (aggressive and withdrawn) syndrome groups showed distinctive response patterns that could not be derived from syndrome scores, with externalizers showing aggression in response to aversive events, and mixed boys showing contextually anomalous responses to positive events. The externalizing and mixed groups each consisted of functionally distinct subgroups that were not detected using syndrome measures. The results clarify how syndromal measures can confuse psychologically distinct children and demonstrate the utility of a teacher-report method that is efficient to administer and sensitive to contextual influences. PMID- 11495176 TI - Cognitive-behavioral treatment for depression in smoking cessation. AB - Cigarette smokers with past major depressive disorder (MDD) received 8 group sessions of standard, cognitive-behavioral smoking cessation treatment (ST; n = 93) or standard, cognitive-behavioral smokiig cessation treatment plus cognitive behavioral treatment for depression (CBT-D; n = 86). Although abstinence rates were high in both conditions (ST, 24.7%; CBT-D, 32.5%, at 1 year) for these nonpharmacological treatments, no main effect of treatment was found. However, secondary analyses revealed significant interactions between treatment condition and both recurrent depression history and heavy smoking ( > or =25 cigarettes a day) at baseline. Smokers with recurrent MDD and heavy smokers who received CBT-D were significantly more likely to be abstinent than those receiving ST (odds ratios = 2.3 and 2.6, respectively). Results suggest that CBT-D provides specific benefits for some, but not all, smokers with a history of MDD. PMID- 11495177 TI - Appraisals of controllability and coping in caregivers and HIV+ men: testing the goodness-of-fit hypothesis. AB - The authors examined goodness of fit between controllability appraisals and coping in 82 HIV+ and 162 HIV-gay men experiencing the chronic stress of caregiving and 61 HIV+ gay men who were not caregiving. Multiple assessments of each individual over a 2-year period allowed replication of prior cross-sectional research examining goodness of fit, as well as the creation of intraindividual goodness-of-fit scores that were then used to examine within-person fluctuations in goodness of fit over time and goodness of fit as an individual difference variable related to adjustment. Results indicate that the importance of goodness of fit varies for different kinds of coping: The concept was supported for problem-focused coping and, to a lesser extent, for emotion-focused coping but not for meaning-focused coping. Within-subject variation in goodness of fit was related to within-subject variation in depressed mood, but between-subjects variation (individual differences) in fit was not associated with depressed mood, suggesting that goodness of fit is better understood as a transactional variable than as a personality or "coping style" variable. PMID- 11495178 TI - Does competitive employment improve nonvocational outcomes for people with severe mental illness? AB - The authors examined the cumulative effects of work on symptoms, quality of life, and self-esteem for 149 unemployed clients with severe mental illness receiving vocational rehabilitation. Nonvocational measures were assessed at 6-month intervals throughout the 18-month study period, and vocational activity was tracked continuously. On the basis of their predominant work activity over the study period, participants were classified into 4 groups: competitive work, sheltered work, minimal work, and no work. The groups did not differ at baseline on any of the nonvocational measures. Using mixed effects regression analysis to examine rates of change over time, the authors found that the competitive work group showed higher rates of improvement in symptoms; in satisfaction with vocational services, leisure, and finances; and in self-esteem than did participants in a combined minimal work-no work group. The sheltered work group showed no such advantage. PMID- 11495179 TI - Psychotherapy research with ethnic minorities: empirical, ethical, and conceptual issues. AB - There is an increasing demand for psychotherapy among ethnic minority populations. Yet, there is not adequate evidence that empirically supported therapies (ESTs) are effective with ethnic minorities. Ethical guidelines suggest that psychotherapies be modified to become culturally appropriate for ethnic minority persons. Conceptual approaches have identified interdependence, spirituality, and discrimination as considerations for culturally sensitive therapy (CST). However, there is no more empirical support for the efficacy of CSTs than there is for the efficacy of ESTs with ethnic minority populations. The chasm between EST and CST research is a function of differences between methods and researchers in these 2 traditions. Specific recommendations for research collaboration between CST and EST researchers are offered. PMID- 11495180 TI - Influences of gender and weight gain on short-term relapse to smoking in a cessation trial. AB - Few researchers have studied whether weight gain has an impact on short-term relapse to smoking. The authors of this study investigated predictors of relapse among 989 participants (60% women) in a randomized, double-blind, 10-week multicenter trial to determine the effect of fluoxetine (30 or 60 mg) versus placebo in combination with behavioral counseling for smoking cessation. Medication compliance and smoking status were biochemically verified. At Visit 2, participants were asked to set a quit date within the subsequent 2 visits. A proportional hazards regression model was used to predict risk of relapse within the first 3 months of quitting. Weight gain predicted relapse, but for men only. Female gender also predicted relapse. The results led the authors to question whether postcessation weight gain interventions should be restricted to women smokers. PMID- 11495181 TI - Does smoking abstinence self-efficacy vary across situations? Identifying context specificity within the Relapse Situation Efficacy Questionnaire. AB - According to self-efficacy and relapse theories. abstinence self-efficacy (ASE) ratings should be context-specific: they should vary across situations. This variability may be important, as it could signal high-risk for relapse situations. In this study, confirmatory factor analysis was used to identify situational variability in a novel ASE assessment (Relapse Situation Efficacy Questionnaire. or RSEQ). Results supported a hierarchical structure, where both context-specific and unidimensional measures of ASE exist within the assessment. Context-specific factors included Negative Affect, Positive Affect, Restrictive Situations (to smoking). Idle Time, Social-Food Situations, Low Arousal, and Craving. Multiple context-specific factors and the aggregate factor predicted cessation outcome, even after controlling for concurrent smoking rate. However, the context rated with the least confidence proved to be the best outcome predictor, suggesting the existence of "Achilles' heel" situations. These data indicate the internal and predictive validity of the RSEQ. PMID- 11495182 TI - Correlates of intimate partner violence among male alcoholic patients. AB - Male-to-female partner violence was investigated in heterosexual couples with an alcoholic male partner. Partner violent (PV) alcoholic patients (n = 183), when compared with nonviolent (NV) alcoholic patients (n = 120), had more antisocial personality characteristics, greater alcohol problem severity, greater use of other drugs, higher relationship distress, and stronger beliefs in the link between alcohol consumption and relationship problems. Demographic factors did not account for these PV-NV differences. Relationship distress and alcohol problem severity had independent associations with partner violence. Relationship adjustment and drug use remained significantly associated with partner violence, whereas alcohol problem severity did not, after controlling for patient antisocial traits. Beliefs in the link between drinking and relationship problems were associated with partner violence independent of other clinical factors. PMID- 11495183 TI - Posttraumatic stress disorder arising after road traffic collisions: patterns of response to cognitive-behavior therapy. AB - Road traffic collisions (RTCs) are common precipitants of posttraumatic stress disorder (PTSD). Two preliminary studies suggest that cognitive-behavior therapy (CBT) is, on average, effective in treating this disorder, although the major patterns of treatment outcome remain to be identified. Such outcomes might include treatment response, partial response, and response followed by relapse. To identify these patterns. 50 people with RTC PTSD completed a 12-week course of CBT, with outcome assessment extending to 3-month follow up. Dynamic cluster analyses revealed 2 replicable patterns of outcome: one for responders (n = 30) and one for partial responders (n = 20). Partial responders, compared with responders, tended to have more severe pretreatment numbing symptoms and greater anger about their RTC, along with lower global levels of functioning, greater pain severity and interference, and greater depression and were more likely to be taking psychotropic medications. Responders and partial responders did not differ in homework adherence, number of sessions attended, therapist effects, or stressors occurring during therapy or in the presence or absence of RTC-related litigation. Implications for enhancing treatment outcome are discussed. PMID- 11495184 TI - Evaluating a prevention program for teenagers on sexual coercion: a differential effectiveness approach. AB - The authors evaluated a coeducational program for teenagers on preventing sexual coercion in dating situations. Students examined individual and social attitudes underlying coercive sexual behavior and learned communication skills aimed at preventing or dealing with unwanted sexual advances. Instruction was enhanced by video and an interactive video "virtual date." Outcomes were assessed using sexual attitude scales with a sample of 458 high school students. Student health education classes were randomly assigned to either a treatment or a control condition. Findings, based on a latent variable model of differential effectiveness, showed that students in the treatment group with initial coercive attitude scores at or above the mean benefited significantly more than students with the same range of scores in the control group. PMID- 11495185 TI - Response to cognitive therapy in depression: the role of maladaptive beliefs and personality disorders. AB - This study examined whether personality disorder status and beliefs that characterize personality disorders affect response to cognitive therapy. In a naturalistic study, 162 depressed outpatients with and without a personality disorder were followed over the course of cognitive therapy. As would be hypothesized by cognitive theory (A. T. Beck & A. Freeman, 1990), it was not personality disorder status but rather maladaptive avoidant and paranoid beliefs that predicted variance in outcome. However, pre- to posttherapy comparisons suggested that although patients with or without comorbidity respond comparably to "real-world" cognitive therapy, they report more severe depressive symptomatology at intake and more residual symptoms at termination. PMID- 11495186 TI - Categorizing partner-violent men within the reactive-proactive typology model. AB - A system for categorizing partner-violent men as either reactive or proactive aggressors was developed and evaluated in the present study. Sixty partner violent men were reliably categorized, and the distribution (62% reactive, 38% proactive) fell within the expected range. Some construct validity was demonstrated, as several significant predicted group differences were found on factors of theoretical relevance to the typology model (affectivity, personality, and violence in the family-of-origin). Proactively versus reactively categorized participants were (a) more dominant and less angry during a 10-min interpartner interaction, (b) more antisocial and aggressive-sadistic and less dependent, and (c) more frequently classified as psychopathic (17% vs. 0%). Research and clinical implications of the system are discussed, as is the potential overlap between the reactively and proactively categorized partner-violent men in this study with previously identified types. PMID- 11495187 TI - Changing patterns of inpatient care for children and adolescents at the Menninger Clinic, 1988-1994. AB - The authors describe trends in inpatient psychiatric length of stay (LOS) and admissions for the population of children and adolescents (N = 784) at the Menninger Clinic from 1988 to 1994. During this period. median LOS declined dramatically from 7 months to 3 weeks, whereas admissions increased 4-fold. The diagnostic case mix changed substantially, with a crossover in modal principal diagnosis from personality disorder to affective disorder. Use of medications became almost universal. Diagnosis and medication use became less important determinants of LOS over time. The practical implications of these patterns include higher patient turnover, fewer inpatient clinical contact hours, and heightened importance of continuity with outpatient care. Research should center on the impact of declining LOS on clinical and functional outcomes for children and adolescents. PMID- 11495188 TI - Economic globalization and oral health. AB - OBJECTIVES: To briefly review the origins of economic globalization and examine the evidence available concerning its possible impact on oral health. DESIGN: Based on Medline searches 1966-1999 and review of Health Wrights: Politics of Health database. SPECIFIC ORAL DISEASES: Dental caries, destructive periodontal diseases, cancrum oris and oral cancer. CONCLUSIONS: The reported growing disparity between rich and poor populations, both internationally and nationally, is arguably being exacerbated by economic globalization. Increasing levels of the above specific oral diseases might be attributed, in part, to this economic phenomenon. PMID- 11495189 TI - Iron and vitamin deficiencies, endocrine and immune status in patients with primary Sjogren's syndrome. AB - OBJECTIVES: To study the prevalence of iron and vitamin deficiencies, endocrine disorders and immunological parameters in patients with primary Sjogren's syndrome (1 degree SS). DESIGN AND SUBJECTS: At the time of the establishment of the diagnosis of 1 degree SS in 43 consecutive patients, a clinical examination including haematological analyses was performed. The patients' medical records were also reviewed. SETTING: Patients referred for diagnosis to The University Hospital, Linkoping, a secondary or tertiary referral hospital serving the middle part of southern Sweden. RESULTS: In total, current or previously treated iron and vitamin deficiencies were registered for 63% of the 1 degree SS patients (iron 51%, vitamin B12 25%, folate 9%). Current low ferritin was noted in 24%, low iron saturation in 37%, decreased vitamin B12 in 13% and folate in 9%. Thyroid disease was found in a total of 33% and 30% had had autoimmune thyroiditis. Three patients (7%) had verified diabetes mellitus. Erythrocyte sedimentation rate (ESR) was raised in 65% of the patients and 84% had a polyclonal increase of Ig. Rheumatoid factor (RF) was detected in 85%, antinuclear antibody (ANA) in 74%, anti-SS-A in 88% and anti-SS-B in 73% of the patients. CONCLUSION: Iron and vitamin deficiencies and thyroid diseases are common in patients with 1 degree SS. Since these disorders often are treatable and may affect the patients' distress as well as their immune and exocrine function, an active, recurrent search for deficiencies, endocrine diseases and other frequently recorded disorders is recommended. PMID- 11495190 TI - Seasonal variation of acute necrotising ulcerative gingivitis in South Africans. AB - OBJECTIVES: To examine seasonal variations in the presentation of acute necrotising ulcerative gingivitis (ANUG) in a previously unstudied population in Cape Town, South Africa. DESIGN: A descriptive study of all presenting cases of ANUG at a health center over 6 years; patients were examined at one point in time. SETTING: A clinic-based cross-sectional study was carried out among patients presenting routinely to the oral medicine clinic at the University of the Western Cape. Patients at this clinic were mostly colored and black people, and were of low socioeconomic status. SUBJECTS AND METHODS: All patients presenting to the periodontal clinic were examined during the period from 15 March 1992 to 15 March 1998 and interviews were conducted. A comprehensive clinical intra-oral and peri-oral examination was carried out using a dental mirror and blunt probe by three trained calibrated dental examiners. The minimum criteria for inclusion as an ANUG patient were painful, bleeding gums with irreversible destruction of the interdental papillae (cratering) and marginal gingiva. Patients with multiple small ulcerations not restricted to the gingiva, but including other oral sites such as the tongue and buccal mucosa were diagnosed as herpetic stomatitis. Other distinguishing characteristics of ANUG included presence of a pseudomembrane, odor, a strange taste in the mouth, and hypersalivation. Swabs and biopsies taken to verify the diagnosis when necessary. If there was any doubt as to the diagnosis of the lesion it was not included among the ANUG cases. MAIN OUTCOME MEASURE: Presence of ANUG. RESULTS: Among the 19944 patients presenting for periodontal treatment, 684 were found to have ANUG. The ANUG patients consisted of 58% males; 73% of the ANUG patients were between the ages of 5 to 12 years. The majority of ANUG patients presented during the summer (55.4%), 27.7% in autumn and 8.4% during both winter and spring. Significant differences were evident between the numbers seen during the different seasons. CONCLUSIONS: This study demonstrated a significant seasonal variation in the occurence of ANUG in Cape Town, South Africa. PMID- 11495191 TI - Benign fibro-osseous lesions of the jaw bones in Jamaica: analysis of 32 cases. AB - OBJECTIVE: It is intended to analyse all the cases of benign fibro-osseous lesions seen at the two major public hospitals in Jamaica over a 15-year period and to compare our result with what has been previously documented by other authors. No such analysis has been carried out in Jamaica or elsewhere in the English-speaking Caribbean. MATERIALS AND METHODS: The case files of patients histologically diagnosed to have a fibro-osseous lesion at both the Kingston Public Hospital and the Cornwall Regional Hospital in Jamaica from 1980 to 1995 were retrieved and information about these lesions was documented. The WHO classification for benign fibro-osseous lesions was used. RESULTS: A total of 32 cases of benign fibro-osseous lesions were recorded over the 15-year period. Of these, 15 were histologically reported as fibrous dysplasia, 10 as ossifying fibroma, three as gigantiform cementoma, one each as periapical cemental dysplasia and cementoblastoma, and two cases of recurrent fibrous dysplasia. With the exception of gigantiform cementoma the age at presentation for these lesions is not consistent and this can be particularly noted for ossifying fibroma and fibrous dysplasia, both of which occurred between the age of 10 and 68 years. CONCLUSION: These lesions are more common in females, furthermore it is suggested that the inconsistent age at presentation for some of these lesions may be peculiar to Jamaica, and may be due to delay in reporting a lesion by the patient, as routine dental examination is not a common practice. PMID- 11495192 TI - Craniofacial tissues including tooth buds in fetal hypohidrotic ectodermal dysplasia. AB - OBJECTIVE: Hypohidrotic ectodermal dysplasia (HED) comprises defects in hair, teeth, and sweat glands. Disturbances in other ectodermal tissues have been associated with the condition. Our objective was to examine ectodermal craniofacial structures histologically in a fetus with HED and to compare the findings to similar structures in normal control fetuses. MATERIALS AND METHODS: A male fetus diagnosed with HED was therapeutically aborted in the 15th week of gestation. One male and two female healthy fetuses were used as normal controls. All fetuses were examined with parental consent, and had comparable sizes. Their bone maturation stage in the hand was identical. Tissue blocks from the craniofacial region were excised from all fetuses and prepared for histological analysis (formalin fixed, stained with toluidine blue or Alcian blue). The tissues examined were: tooth buds, skin and skin appendages, oral mucosa including minor salivary glands, major salivary glands, lacrimal glands, and adenohypophysis. RESULTS: Fewer tooth buds, minor salivary glands, and hair follicles were observed in the HED fetus as compared to controls. The structures of the epidermal components in the developing HED organs were loose and disorganised. The adhesion between the ectodermal and mesenchymal organ components in the HED fetus seemed to be disturbed. PMID- 11495193 TI - Immunohistochemical detection of amelogenin and cytokeratin 19 in epithelial odontogenic tumors. AB - OBJECTIVE: Epithelial odontogenic tumors exhibit considerable histological variation and are classified into several benign and malignant entities. Expression of amelogenin and cytokeratin 19 (CK19), that are potentially useful polypeptides for identification of odontogenic epithelial components, was evaluated in various types of epithelial odontogenic tumors. MATERIALS AND METHODS: Specimens of 33 ameloblastomas, three calcifying epithelial odontogenic tumors (CEOTs), two clear cell odontogenic tumors (CCOTs) and five malignant ameloblastomas were examined by immunohistochemistry using anti-amelogenin and anti-CK19 antibodies. RESULTS: Immunohistochemical reactivity for amelogenin was detected in many peripheral columnar or cuboidal cells and some central polyhedral cells in ameloblastomas, and histological variants showed various degrees of amelogenin expression. Expression of CK19 was diffusely present in neoplastic cells in ameloblastomas, and decreased expression was found in keratinizing cells of acanthomatous variants and some neoplastic cells of desmoplastic variants. In CEOTs, immunohistochemical reactivity for amelogenin was detected in neoplastic cells and intercellular amyloid-like materials, whereas CK19 was expressed in neoplastic cells. CCOTs showed positive reactivity for amelogenin and CK19 in neoplastic cells. Malignant ameloblastomas exhibited various degrees of amelogenin expression with constant CK19 expression in neoplastic cells. CONCLUSION: Diverse types of epithelial odontogenic tumors express amelogenin and CK19, suggesting that these tumors have ameloblastic differentiation or odontogenic epithelial properties. PMID- 11495194 TI - Maryland dental hygienists' knowledge and opinions of oral cancer risk factors and diagnostic procedures. AB - OBJECTIVES: To examine Maryland dental hygienists' (DHs) knowledge of oral cancer risk factors and diagnostic procedures, as well as opinions about the currency and adequacy of their oral cancer knowledge, educational preparation, interest in, and preferred types of, continuing education courses. METHODS: Seven hundred DHs were randomly selected from a registry of 2677 Maryland licensed dental hygienists. A mailed survey instrument provided baseline data on 331 (RR = 60%) DHs. Stratified tabular and logistic analytical techniques were employed (alpha level < or =0.05). RESULTS: Most correctly identified tobacco use (99.7%) and alcohol use (89%) as risk factors; however, 31% incorrectly identified poor oral hygiene as a risk factor. 64% of DHs correctly identified older age as a risk factor, yet only 16% identified that the majority of oral cancers are diagnosed in the 60+ year old age group. Nearly 91% correctly identified the examination procedures of the tongue for oral cancer detection; while only 16% of DHs correctly identified erythroplakia and leukoplakia as the conditions most likely associated with oral cancer. CONCLUSIONS: Gaps in knowledge exist and strongly suggest the need for continuing education courses to clarify risk factors and diagnostic procedures associated with earlier oral cancer detection and prevention. PMID- 11495195 TI - A model for internal derangement and osteoarthritis of the temporomandibular joint with experimental traction of the mandibular ramus in rabbit. AB - OBJECTIVES: The purpose of this study is to make a model of internal derangement (ID) and osteoarthritis (OA) by postero-superior displacement of the mandible in the rabbit temporomandibular joint (TMJ). MATERIALS AND METHODS: Sixteen Japanese white rabbits (8 months old) were subjected to traction of the mandibular ramus in the postero-superior direction unilaterally using elastic force and five rabbits were used as the control. The operations were performed without surgical invasion of the TMJ capsule. Temporomandibular joint tissue was removed after 4 weeks and Technovit and paraffin sections with or without decalcification were observed histologically. RESULTS: In contrast to the control group, it showed anterior disc displacement, disc deformity, resorption of subchondral bone and osteophyte formation in the drawing side of the experimental group. Intra articular fibrous adhesions were observed in both sides. CONCLUSION: It is concluded that posterosuperior displacement of the mandible caused ID and OA in the rabbit TMJ. This model may contribute to the understanding of the etiology of human temporomadibular disorders (TMD). PMID- 11495196 TI - New local hyperthermia using dextran magnetite complex (DM) for oral cavity: experimental study in normal hamster tongue. AB - The possibility of dextran magnetite complex (DM) as a new hyperthermic material was examined in this study. DM suspension of 56 mg ml(-1) iron concentration was locally injected into the normal tongue of golden hamster. DM injected tongues were heated by 500 kHz alternating current (AC) magnetic field and its serial changes in temperature were recorded at 30-s intervals. The temperature of DM injected tongue was maintained at about 43.0-45.0 degrees C for 30 min by changing the AC magnetic field intensity. While temperature elevations of the contralateral tongue and the rectum were only of minor degree. In experiment on the extent of heating area, there was correlation between volume of black stain area and amount of the injected DM suspension (Y = - 18.1 + 1.94X, r = 0.931, P < 0.0001, n = 9 ). Histological examination after heating revealed brown uniform DM accumulation in the connective tissue between fibers of the tongue muscle. Except for vascular dilatations, no tissue damage was seen in the heated tongue. Thus, DM which has the possibility of selective and uniform heating in local hyperthermia might be useful for oral cancer therapy. PMID- 11495197 TI - Intraoral myxoid nerve sheath tumour. AB - A case of an intraoral myxoid nerve sheath tumour of the dorsum of the tongue in a 73-year-old Caucasian male is reported. This case describes the oldest patient with this pathology to date. Immunoperoxidase staining for neuron-specific enolase (NSE) and epithelial membrane antigen (EMA) expression demonstrated the perineural origin of the lesion. PMID- 11495198 TI - Oro-dental and craniofacial anomalies in LEOPARD syndrome. AB - 'LEOPARD syndrome' is a syndrome affecting many systems or organs. The main anomalies are summarized in the acronym LEOPARD in which each letter corresponds to the damage of a given organ. In the presented case, there are oro-dental and craniofacial anomalies. The relationship between the LEOPARD syndrome and the given anomalies and the possibilities of the management and the follow-up of the diseased are studied. The clinical case presented is a 4-year-old boy with LEOPARD syndrome showing retardation of dental development, delayed development and possible agenesis of permanent teeth, and craniofacial anomalies (osseous hypodevelopment). The bibliographical study shows that LEOPARD syndrome is due to damage of the neural crest cells. Thus, the dental and craniofacial anomalies arise since neural crest cells participate in the formation of the teeth and some craniofacial bones. Therefore, dental and craniofacial anomalies might be expected in some cases of the disease. The therapeutic management of the lesions, and the follow-up of the patient would be done by a multidisciplinary team. PMID- 11495199 TI - CD4 lymphocyte percentage vs CD4 lymphocyte count as an immunological marker of oral candidiasis in HIV-infected patients. PMID- 11495200 TI - The regulation of chiropractic practice in Europe. AB - Non-conventional medicine, including chiropractic, is now in wide use throughout Europe but is often unregulated. The purpose of this paper is to provide an overview of present and future regulation of chiropractic practice in Europe. Companion papers look at specific legal and professional issues in Finland, where chiropractors are regulated by law, and France, where they are not. PMID- 11495201 TI - Cancer genetics: a model for multifactorial conditions? AB - Research in the area of genetic susceptibility and cancer is beginning to challenge traditional socio-ethical and legal norms. In particular, increased personal data protection legislation is severely constraining the ability to maintain or initiate new cancer registries for proper epidemiological purposes. Likewise, the principle and obligation of the confidentiality of genetic information cannot remain sacrosanct in the face of the immediate health needs of biological relatives. Finally, participation in research and even the willingness to be tested and treated is constantly threatened by the uncertainty surrounding insurability. What are the new ethical parameters emerging in research in cancer genetics? PMID- 11495202 TI - Legal issues in the development and use of clinical practice guidelines. AB - Over the last ten years, the development and dissemination of practice guidelines has increased at a rapid pace. From a legal point of view, it should always be made clear whether a guideline has been developed to improve the quality of care and is based on medical evidence and professional experience, or whether other concerns and considerations (organisational, financial) did prevail. Guidelines should not simply be imposed on health professionals; that would result in a standardisation of care that does no justice to individual patient needs and preferences. Patients have the right to be informed about reasonable and realistic treatment alternatives, even if they are not included in the guideline. Using cost effectiveness analysis in guideline development can help to reduce care of dubious effectiveness. But if cost considerations are used as a reason to limit effective medical care, the guidelines in question need political legitimation. PMID- 11495203 TI - Bio-libraries and DNA storage: assessment of patient perception of information. AB - BACKGROUND: Since the creation of places where DNA is stored, a number of questions appear about protecting the individual, confidentiality and the validity of the proposed consent to storage. This study, originating in France, analyzes and assesses the impact of the information given by the clinician to the patient regarding having his DNA stored. How does the patient receive the information? What is left of it? Is he informed enough to sign a "well-informed" consent? METHODS: A questionnaire issued to 170 patients in the same department was used to assess: (1) their knowledge of genetics, (2) their knowledge of DNA storage, (3) their perception of the consent. FINDINGS: A third of the patients answered, 20% of whom understood the objectives of a genetic test. None of the patients contacted was aware that he had samples stored in a DNA bank, that genetic analyses had been undertaken or remembered that he had signed a consent (which indeed exists). INTERPRETATION: The idea of a "well-informed" consent belongs to the criteria defining the validity of a cure and research agreement. Yet, its validity is doubted as soon as the information it implies is not understandable by the patients. Indeed, can the individual be considered autonomous, in the philosophical sense, in a well-informed decision making if the information given to the patient has not been well understood? PMID- 11495204 TI - Informed consent, informed refusal, informed choice--what is it that makes a patient's medical treatment decisions informed? AB - Informed consent is the fundamental ethical and legal doctrine that protects the patient's rights of personal autonomy and bodily self-determination. An adjunct to the doctrine of informed consent advanced by some is the notion of informed refusal. According to advocates of this concept, incoherent, unconscious, or otherwise incapacitated patients cannot make informed treatment choices because such patients cannot receive a full and current explanation of their health problems and treatment options. This, in turn, raises serious questions about what it is that makes a patient's treatment decisions--whether consents or refusals--informed. Is current, detailed information about the patient's medical condition and treatment options an ethical and legal prerequisite? Can non medical values and concerns of the patient ever suffice to make the patient's treatment choices informed? How does the concept of informed refusal affect the use of health-care advance directives? This paper will address these important questions. PMID- 11495205 TI - Medical confidentiality after a patient's death, with particular reference to The Netherlands. AB - This article concerns medical confidentiality after a patient's death. At international level there is an agreement that medical data of deceased patients should continue to be treated as confidential and therefore be protected by law. In the Netherlands, courts have ruled along the same lines. After death, the physician is bound to confidentiality and if necessary should invoke his right to remain silent. However, it is also accepted by the courts that circumstances can occur in which a doctor may disclose confidential information to third parties such as relatives. Disclosure of medical data is justified when the agreement of the deceased can be presupposed, or (if this is impossible) if there are such important interests of third parties involved that the obligation to maintain confidentiality may be put aside. According to present Dutch jurisprudence, confidentiality of medical data is regarded as a personal right that is not inherited by the relatives. Whether the position of the latter should be reinforced by introducing statutory provisions, is to be further explored. PMID- 11495206 TI - Compulsory medication within the prison system. AB - This paper deals with: 1. the legal principles concerning compulsory medication of the mentally disordered prisoner; 2. the legal possibilities and limitations for compulsory medication within penitentiaries in the Netherlands; 3. a critical evaluation of these legal rules with a view to the above-mentioned principles. PMID- 11495207 TI - Further offences by discharged psychiatric patients--can somebody be held liable? AB - The decision to release a state patient who is unfit to stand trial and/or not criminally responsible, who had been committed to a mental institution in terms of the Criminal Procedure Act or who was held in terms of section 28 of the Mental Health Act, may have harmful or even fatal consequences. The multi professional team at Oranje Hospital will only institute discharge procedures regulated in terms of section 29 of the Mental Health Act when it is of the opinion that a state patient's mental illness is stable and well-controlled and his clinical picture is such that he qualifies to be discharged. In this discussion focus will be placed on: a. the two discharge procedures applied in South Africa. b. four case studies which will be presented dealing with the observation period, admission, offence, diagnosis, treatment, hospitalisation, leave periods, discharge on certain conditions, re-admission and further offences. c. the question of whether the State may be held liable for the negligent or wrongful discharge or release of a state patient who subsequently commits a serious crime. PMID- 11495208 TI - Treatment refusal in the involuntarily hospitalized psychiatric population: Canadian policy and practice. AB - Policies governing coercive psychiatric interventions must be scrutinized to ensure they are not only beneficent but just. Within Canada, in seven of ten provinces, persons may be detained in hospital against their will for dangerousness due to mental disorder but, if capable of making treatment decisions, may refuse treatment for the very illness that led to detention. In examining this issue, the purpose of this paper is threefold. First, it will demonstrate that treatment refusal amongst involuntarily hospitalized psychiatric patients has arisen from a recent separation, in law, of hospitalization from treatment. Second, it will argue that legal provisions that allow the detention of psychiatric patients prior to assessment of their treatment decision-making capacity discriminate against the mentally ill and undermine health service delivery. Finally, this paper will propose that a more just policy would allow involuntary hospitalization of only those patients who are both incapable of making treatment decisions and harmful to themselves or others. PMID- 11495209 TI - Has Israel and the Russian Federation legislation a full set of laws securing the protection of mental patients relative to United Nations' proposals and do they require modification? AB - This paper compares sections of laws dealing with the care and protection of mental patients in Israel and in the Russian Federation and matches them with the 24 recommendations on the subject proposed by the United Nations (UN) in 1988. Particular attention is given to issues related to forensic and ethical subjects. The UN proposals constitute a most laudable reform-inducing guideline for global mental health care systems, especially because of their emphasis on the preservation of mental patients' basic human rights. The proposals' strength springs from their influence regarding the prevention of misuse of psychiatry with focus on the care of inpatients and control over executive agencies. The detailed analysis of the similarities between the Israeli and Russian legislative approach reveals some badly matched organizational structure of many articles in comparison with the UN mandate. Forensic and ethical concepts present in the UN 1988 proposals are well addressed in both the Israeli and Russian mental health legislation. We conclude that the 1991 UN recommendations would demand further modification of the national standards. PMID- 11495210 TI - Reproductive health services and the law and ethics of conscientious objection. AB - Reproductive health services address contraception, sterilization and abortion, and new technologies such as gamete selection and manipulation, in vitro fertilization and surrogate motherhood. Artificial fertility control and medically assisted reproduction are opposed by conservative religions and philosophies, whose adherents may object to participation. Physicians' conscientious objection to non-lifesaving interventions in pregnancy have long been accepted. Nurses' claims are less recognized, allowing nonparticipation in abortions but not refusal of patient preparation and aftercare. Objections of others in health-related activities, such as serving meals to abortion patients and typing abortion referral letters, have been disallowed. Pharmacists may claim refusal rights over fulfilling prescriptions for emergency (post-coital) contraceptives and drugs for medical (i.e. non-surgical) abortion. This paper addresses limits to conscientious objection to participation in reproductive health services, and conditions to which rights of objection may be subject. Individuals have human rights to freedom of religious conscience, but institutions, as artificial legal persons, may not claim this right. PMID- 11495211 TI - Assisted human reproductive techniques--emerging ethical and legal implications. AB - The unprecedented and rapid advances in medical sciences have revolutionized modern medicine and surgery in a number of ways. These advances such as human embryo cloning, trans-genetic manipulation, in-vitro fertilization and surrogate motherhood have been able to cure infertility to a large extent by offering 'made to measure or super babies' and can cure a number of conventional diseases and improve the quality of life. Scientists and sociologists are concerned about moral and ethical issues, which have arisen due to these advances in medicine. The new biotechnology and the achievements in bio-engineering offered hope to those couples in fulfilling their yearning for children. What we witness today is just the beginning and not the end of the adventurous march of science in unravelling the mysteries of nature. The moot point now is how can nature be engineered to yield positive results without offending values of human morality, ethics and decency. It is high time that governments and society take stock of the current dilemma, as these advances in biosciences pose a threat to the moral and ethical fabric of modern society. PMID- 11495212 TI - Dutch alcohol policy developments: the last decades and present state of affairs. AB - The article addresses the Dutch alcohol policy over the last decades and the present state of the art. In Dutch society an ambivalent attitude towards alcohol consumption prevails. Developments in legislation and regulations, in prevention and care are described. Several key problems are identified. Among others it is concluded that too little attention is paid to alcohol compared to other drugs. Both in prevention and care evidence-based work has to be stimulated. Legislation and regulations are insufficiently used and evaluated. The government needs to constantly evaluate its policy and develop its options within the area of legislation and regulations, which should also include a critical comparison with the situation in other countries. PMID- 11495213 TI - Euthanasia: a denial of sense. AB - Is active euthanasia an expression of respect for human dignity and freedom or not? Liberal utilitarians and catholic personalists differ on this question because they use different concepts of freedom and have different attitudes to whether life has a sense beyond material utility. While the former can claim that one has no duty to go on living an unpleasant and useless life against one's will, for the latter even a materially substandard life is a chance for spiritual growth not to be discarded because human suffering is meaningful too, and not a mere senseless disutility. Patient autonomy will not, for a personalist doctor, imply a duty to fulfil the patient's wish for euthanasia, because his moral duty towards his own soul overrides all other considerations. The threshold argument, commonly appealed to by personalists against legalizing euthanasia, should not be interpreted as a factual conjecture of the worst possible scenario but rather as a principal claim according to which a state which lets citizens to be reified as means for utility is thereby already entirely immoral. PMID- 11495214 TI - Perspectives on stress in fibromyalgia. PMID- 11495215 TI - Oral contraceptive use is associated with increased cardiovascular reactivity in nonsmokers. AB - Women who smoke and take oral contraceptives (OCs) have significantly increased risk of cardiovascular disease, but the exact mechanismsfor the increased risk are not known. Cardiovascular reactivity to psychological stress may be one mechanism for the enhanced risk, but the small number of studies examining whether OC users who smoke have greater reactivity have produced mixed results. The purpose of this study was to examine the effect of chronic cigarette smoking, acute nicotine administration, and OC use on cardiovascular and lipid reactivity. Sixty healthy women, half of whom had been using OCs for at least the previous 6 months, participated in the study. Approximately two thirds were smokers and were randomized to be tested after either a 12-hr nicotine deprivation or administration of nicotine gum. One third were nonsmokers. Heart rate, blood pressure, and lipid measures were taken at rest, during a videotaped speech task, and during recovery from the task. Results indicated that, among OC nonusers, there was no effect of smoking status or nicotine administration on cardiovascular reactivity. However, among OC users, nonsmokers had significantly greater heart rate and diastolic blood pressure reactivity to stress. These data show that acute nicotine administration, in the form of nicotine gum, has no effect on cardiovascular or lipid stress reactivity in women. However OC use among nonsmoking women is associated with greater cardiovascular reactivity to stress. PMID- 11495216 TI - Appraised control, coping, and stress in a community sample: a test of the goodness-of-fit hypothesis. AB - Lazarus and Folkman proposed one of the most comprehensive theories of stress and coping in the psychology literature, but many of their postulates have received little empirical attention, and some of the existing research hasyielded contradictory findings. This longitudinal study sought to clarify the associations among control appraisal, coping, and stress within this theoreticalframework. The theory postulates that coping strategies used tend to match the level of appraised controllability of the stressor (matching hypothesis). It further states that the effects of problem-focused versus emotion focused coping are moderated by the appraised controllability of the stressor (goodness-of-fit hypothesis). An alternative to the latter is the main-effects hypothesis, which states that problem-focused coping is generally more effective in reducing distress regardless of appraisal. These hypotheses were tested on 72 adults who completed questionnaires on coping and control appraisal. Stress was assessed using self-report (Symptom Checklist-90-Revised) and a behavioral measure (proofreading task) at two times approximately 2 months apart. Appraised control significantly predicted type of coping such that greater control was associated with more problem-focused and less emotion-focused coping. Although the main-effects hypothesis was not supported, the goodness-of-fit hypothesis was partly confirmed by a significant control by emotion-focused coping interaction predicting both self-report and behavioral measures of stress. PMID- 11495217 TI - Illness episodes and cortisol in healthy older adults during a life transition. AB - Alterations in neuroendocrine functioning and in the neuroendocrine response to stress have been observed in older adults. Stressful life events have also been associated with increased illness vulnerability. However effects of natural life stressors on neuroendocrine functioning and health of the elderly have not been well characterized. This research examines relationships among cortisol, dehydroepiandrosterone sulfate (DHEA-S), distress, and illness episodes in an elderly population experiencing the life transition of housing relocation. Thirty older adults moving to congregate livingfacilities were assessed in their homes 1 month premove and 2 weeks postmove. Twenty-eight nonmoving comparison participants were assessed at similar time points. Assessments included measures of intrusion, cortisol, DHEA-S, and self-reported infectious illness episodes. Movers reported more illness episodes between the two assessments than controls. Significant alterations in neuroendocrine measures were not observed among movers at either time point. Individuals with more intrusive thoughts had higher cortisol levels concurrently and prospectively, but these relationships did not vary by group. Greater intrusion at premove was associated with a greater likelihood of reported illness episodes between the two assessments, but there were no relationships between neuroendocrine factors and illness episodes, and intrusion did not mediate the relationships between group and likelihood of illness. In healthy elders, a temporary life stressor may increase vulnerability to illness but does not PMID- 11495218 TI - Neighborhood problems as sources of chronic stress: development of a measure of neighborhood problems, and associations with socioeconomic status and health. AB - The impact of the residential neighborhood on health and well-being is being increasingly recognized in behavioral medicine, with evidence for neighborhood level effects that are independent of the individual characteristics of residents. This study addressed the possibility that the effects of neighborhood are due in part to exposure to community-wide stressors rather than variations inprotective factors such as social capital. A questionnaire survey including a 10-item neighborhood problems scale and measures of self-reported health, health behaviors, and social capital was completed by 419 residents of 18 higher socioeconomic status (SES) neighborhoods and 235 residents of 19 lower SES neighborhoods. Data were analyzed using regression and multilevel methods. Neighborhood problem scores were greater in lower than higher SES neighborhoods, positively associated with individual deprivation, and negatively correlated with social capital. Neighborhood problems were not related to smoking, diet, alcohol consumption, or physical activity. However, neighborhood problems were associated with poor self-rated health, psychological distress on the General Health Questionnaire, and impaired physical function, independent of age, sex, neighborhood SES, individual deprivation, and social capital. Adjusted odds ratios for the highest versus lowest neighborhood problem quartiles ranged from 2.05 (confidence interval = 1.15-3.69) for poor self-rated health to 3.07 (1.63 5.79) for impairedphysical function. The results provide preliminary evidence that residential neighborhood problems constitute sources of chronic stress that may increase risk ofpoor health. PMID- 11495219 TI - The effects of exercise on the quality of life of frail older adults: a preplanned meta-analysis of the FICSIT trials. AB - The Frailty and Injuries: Cooperative Studies of Intervention Techniques (FICSIT) was a linked series of randomized clinical trials focused on the benefits of exercise in the frail elderly. This article uses covariate-adjusted preplanned meta-analyses of FICSIT data to evaluate the effect of exercise on quality of life (QOL) outcomes (N = 1,733; age = 73.4 +/- 6.1 years). Results indicate that (a) exercise produced a small but significant improvement in the emotional health component of QOL, trended toward an improved social component, and did not effect perceptions of general health; (b) exercise-related joint and muscle stresses did not increase bodily pain; and (c) QOL improvements were independent of changes in physical functioning. We conclude that exercise can improve QOL in the frail elderly but that the magnitude of the improvement is modest in size. The benefits of a meta-analytical approach for documenting efficacy outcomes across different types of interventions are discussed. PMID- 11495220 TI - Self-rated quality of life measures: effect of change to a low-fat, high-fiber, fruit and vegetable enriched diet. AB - The Polyp Prevention Trial (PPT) was a multicenter, randomized clinical trial to determine the effect of a low-fat (20% of energy from fat), high-fiber (18 g/1,000 kcal/day), high-fruit/vegetable (3.5 servings/1,000 kcal/day) eatingplan on the recurrence of large bowel adenomatous polyps. The PPT provided an opportunity to examine the impact of dietary changes on quality of life. At baseline and annuallyfor 4 years, participants in the Quality of Life Substudy of PPT completed a Quality of Life Factors (QF) Questionnaire, a modified Block National Cancer Institute Food Frequency Questionnaire, and a Health and Lifestyle Questionnaire. The 51-item QF Questionnaire assessed changes in nine domains: taste, convenience, cost, self-care, social, health assessment, health belief health action, and life satisfaction. The analysis compared annual changes in domain scores for intervention (n = 194) and control (n = 200) participants. At Year 1, 363 (92%) completed a questionnaire, and 325 (82%) participants completed a Year 4 questionnaire. There were no statistically significant differences between treatment groups in the change from baseline to Year 1 for the convenience, cost, taste, health assessment, and life satisfaction domains. At Year 1, intervention participants rated the self-care (p < .001), health belief (p = .021), and health action (p < .001) domains significantly higher and the social domain significantly lower (p <.001) than control participants. These changes were consistent through Years 2, 3, and 4. This study PMID- 11495221 TI - Women's perceived and partners' reported support for smoking cessation during pregnancy. AB - Studies of the influence of social support on successful smoking cessation have been based on the smoker's perceptions only. In this pilot study of 58 couples, pregnant women who had smoked in the 30 days before pregnancy and their partners reported the positive and negative support for cessation they had received (women) or provided (partners). Mean levels of the women's and partners' perceptions of support were compared, and correlations of the two reports were analyzed while controlling for the effect of the couple's smoking status. Women's and partners' reports were similar except partners reported wanting the women to stop smoking more than women perceived. Women's and partners' perceived negative support were moderately correlated (r approximately equal to .48, p approximately equal to .001). Partner-reported positive support also was associated with women's perceived negative support (r approximately equal to .30, p approximately equal to .03). These relationships remained signif cant after controllingfor partners' and women's smoking status. Generally, partners reported giving more positive and less negative support than women perceived. Results suggest the need for further examination of couples' perceptions of support and the impact on smoking cessation during pregnancy. PMID- 11495222 TI - Vulnerability to stress among women in chronic pain from fibromyalgia and osteoarthritis. AB - In two investigations, we studied vulnerability to the negative effects of stress among women in chronic pain from 2 types of musculoskeletal illnesses, fibromyalgia syndrome (FMS) and osteoarthritis (OA). In Study 1, there were 101 female participants 50 to 78 years old: 50 had FMS, 29 had OA knee pain and were scheduled for knee surgery, and 22 had OA but were not planning surgery. Cross sectional analyses showed that the three groups were comparable on demographic variables, personality attributes, negative affect, active coping, and perceived social support. As expected, FMS and OA surgery women reported similar levels of bodily pain, and both groups scored higher than OA nonsurgery women. However, women with FMS reported poorer emotional and physical health, lower positive affect, a poorer quality social milieu, and more frequent use of avoidant coping with pain than did both groups of women with OA. Moreover, the perception and use of social support were closely tied to perceived social stress only among the FMS group. In Study 2, we experimentally manipulated negative mood and stress in 41 women 37 to 74 years old: 20 women had FMS, and 21 women had OA. Participantsfrom each group were randomly assigned to either a negative mood induction or a neutral mood (control) condition, and then all participants discussed a stressful interpersonal eventfor 30 min. Stress-related increases in pain were exacerbated by negative mood induction among women with FMS but not women with OA, and pain during stress was associated with decreases in positive affect in women with FMS but not women with OA. These findings suggest that among women with chronic pain, those with FMS may be particularly vulnerable to the negative effects of social stress. They have fewer positive affective resources, use less effective pain coping strategies, and have more constrained social networks than their counterparts with OA, particularly those who experience similar levels ofpain. They also seem to experience more prolonged stress-related increases in pain under certain circumstances, all of which may contribute to a lowering of positive affect and increased stress reactivity over time. PMID- 11495223 TI - Similarity searching in large combinatorial chemistry spaces. AB - We present a novel algorithm, called Ftrees-FS, for similarity searching in large chemistry spaces based on dynamic programming. Given a query compound, the algorithm generates sets of compounds from a given chemistry space that are similar to the query. The similarity search is based on the feature tree similarity measure representing molecules by tree structures. This descriptor allows handling combinatorial chemistry spaces as a whole instead of looking at subsets of enumerated compounds. Within few minutes of computing time, the algorithm is able to find the most similar compound in very large spaces as well as sets of compounds at an arbitrary similarity level. In addition, the diversity among the generated compounds can be controlled. A set of 17,000 fragments of known drugs, generated by the RECAP procedure from the World Drug Index, was used as the search chemistry space. These fragments can be combined to more than 10(18) compounds of reasonable size. For validation, known antagonists/inhibitors of several targets including dopamine D4, histamine H1, and COX2 are used as queries. Comparison of the compounds created by Ftrees-FS to other known actives demonstrates the ability of the method to jump between structurally unrelated molecule classes. PMID- 11495224 TI - Use of surface area computations to describe atom-atom interactions. AB - Accessible surface (ASA) and atomic contact (ACA) areas are powerful tools for protein structure analysis. However, their use for analysis purposes could be extended if a relationship between them and protein stability could be found. At present, this is the case only for ASAs, which have been used to assess the contribution of the hydrophobic effect to protein stability. In the present work we study whether there is a relationship between atomic contact areas and the free energy associated to atom-atom interactions. We utilise a model in which the contribution of atomic interactions to protein stability is expressed as a linear function of the accessible surface area buried between atom pairs. We assess the validity of this hypothesis, using a set of 124 lysozyme mutants (Matthews, 1995, Adv Protein Chem, 249-278) for which both the X-ray structure and the experimental stability are known. We tested this assumption for residue representations with increasing numbers of atom types. Our results indicate that for simple residue representations, with only 4 to 5 atom types, there is not a clear linear relationship between stability and buried accessible area. However, this relationship is observed for representations with 6 to 9 atom types, where gross heterogeneities in the atom type definition are eliminated. Finally, we also study a version of the linear model in which the atom- atom interactions are represented utilising a simple function for the buried accessible area, which may be useful for protein structure prediction studies. PMID- 11495226 TI - Discovery of a novel serine protease inhibitor utilizing a structure-based and experimental selection of fragments technique. AB - We report a set of strategies to develop novel ligands (Structure Based and Experimental Selection of Fragments: SbE-SF). First, a docking simulation utilizing DOCK3.5 is performed in order to screen the fragment database, which was generated with the in-house program FRAGMENT++ specifically for docking simulation purposes. Although the affinity of these small molecules (fragments) is expected to be low, the affinity of fragments selected by computation is assayed by experiment to determine which ones can be potent inhibitors. After determining such key fragments. additional fragments are attached to the key ones in order to increase the binding affinity,taking into account the binding modes predicted by computation. This method has been applied to a thrombin inhibitor study, resulting in the discovery of a novel inhibitor exhibiting pIC50 = 7.9. PMID- 11495227 TI - Optimization of a mathematical topological pattern for the prediction of antihistaminic activity. AB - Molecular topology was used to develop a mathematical model capable of classifying compounds according to antihistaminic activity. The equations used for this purpose were derived using multilinear regression and linear discriminant analysis. The topological pattern of activity obtained allows the reliable prediction of antihistaminic activity in drugs frequently used for other therapeutic purposes. Based on the results, the proposed pattern is seemingly only valid for drugs that interact with histamine through competitive inhibition with H1 receptors. PMID- 11495225 TI - A sequence and structural study of transmembrane helices. AB - A comparison is made between the distribution of residue preferences, three dimensional nearest neighbour contacts, preferred rotamers, helix-helix crossover angles and peptide bond angles in three sets of proteins: a non-redundant set of accurately determined globular protein structures, a set of four-helix bundle structures and a set of membrane protein structures. Residue preferences for the latter two sets may reflect overall helix stabilising propensities but may also highlight differences arising out of the contrasting nature of the solvent environments in these two cases. The results bear out the expectation that there may be differences between residue type preferences in membrane proteins and in water soluble globular proteins. For example, the beta-branched residue types valine and isoleucine are considerably more frequently encountered in membrane helices. Likewise, glycine and proline. residue types normally associated with 'helix-breaking' propensity are found to be relatively more common in membrane helices. Three dimensional nearest neighbour contacts along the helix, preferred rotamers, and peptide bond angles are very similar in the three sets of proteins as far as can be ascertained within the limits of the relatively low resolution of the membrane proteins dataset. Crossing angles for helices in the membrane protein set resemble the four helix bundle set more than the general non redundant set, but in contrast to both sets they have smaller crossing angles consistent with the dual requirements for the helices to form a compact structure while having to span the membrane. In addition to the pairwise packing of helices we investigate their global packing and consider the question of helix supercoiling in helix bundle proteins. PMID- 11495229 TI - Migrants' right to health. PMID- 11495230 TI - Abdominal drains: a brief historical review. PMID- 11495228 TI - Predicting sequences and structures of MHC-binding peptides: a computational combinatorial approach. AB - Peptides bound to MHC molecules on the surface of cells convey critical information about the cellular milieu to immune system T cells. Predicting which peptides can bind an MHC molecule, and understanding their modes of binding, are important in order to design better diagnostic and therapeutic agents for infectious and autoimmune diseases. Due to the difficulty of obtaining sufficient experimental binding data for each human MHC molecule, computational modeling of MHC peptide-binding properties is necessary. This paper describes a computational combinatorial design approach to the prediction of peptides that bind an MHC molecule of known X-ray crystallographic or NMR-determined structure. The procedure uses chemical fragments as models for amino acid residues and produces a set of sequences for peptides predicted to bind in the MHC peptide-binding groove. The probabilities for specific amino acids occurring at each position of the peptide are calculated based on these sequences, and these probabilities show a good agreement with amino acid distributions derived from a MHC-binding peptide database. The method also enables prediction of the three-dimensional structure of MHC-peptide complexes. Docking, linking, and optimization procedures were performed with the XPLOR program [1]. PMID- 11495231 TI - Meningococcal disease--management of serogroup C clusters in a hyperendemic area. AB - Recent evidence in Ireland highlights an increased incidence of meningococcal disease, with group B the prominent serogroup. However, in developed countries, 'clusters 'are predominantly group C. To retrospectively analyse all patients with meningococcal disease in the North Eastern Health Board (NEHB) Region (circa 300,000 population) from January 1997 to December 1998 inclusively, and to highlight the occurrence of two 'clusters' of Group C meningococcal disease, one in Drogheda and the other in Monaghan (both of whom received national media attention). We studied clinical, laboratory (including PCR) and epidemiological data on all patients in the NEHB with a diagnosis of meningococcal disease. Blood for PCR (Polymerase Chain Reaction) was sent to the National Meningococcal Reference Laboratory on all suspected cases. Details of two 'clusters' were studied in depth from an epidemiological perspective. 131 patients with meningococcal disease were notified, with three deaths of 53 notifications in 1997 (7.5% mortality) and six deaths of 78 notifications in 1998 (9.8% mortality). The rate of meningococcal disease per 100,00 in 1997 was 13.1 and rose to 18.6 in 1998. This contrasts with a national average of 8.5 per 100,00 for the Republic of Ireland in 1997. Age peaks occurred in those under one year old and between 14-16 years old, with seasonal peaks in the winter and spring months. In 1997, 57.5% were group B and 35% were group C, with 7.5% not grouped. In 1998, 60.3% were group B and 39.7% were group C. There were two linked cases in Drogheda, with both attending a local national school. Both were confirmed group C disease. In Monaghan, in January 1998, a cluster of four cases of group C disease occurred over a 48 hour period. All were males, ranging in age from 11-16 years. Of the four, one died and one received intensive care treatment in a children's hospital. The cluster received intense local and national media attention. One week later, a fifth case of group C disease occurred in Monaghan and this 10 year old boy made an uneventful recovery. The high endemic rate of meningococcal disease is striking, with a four fold rise over the past five years. Clusters of serogroup C meningococcal disease will be of historical interest now that new conjugate C vaccines are incorporated into the vaccination program. PMID- 11495232 TI - The effects of age on accident severity and outcome in Irish road traffic accident patients. AB - Road traffic accidents (RTA's) are the leading cause of fatal trauma in Ireland. Although older drivers are the safest group of drivers in the population, once involved in a crash they are more likely to sustain a severe injury or death. The experiences of Irish elderly RTA victims has not been previously documented. We studied older RTA patients admitted to two Irish trauma centres in 1995. Of 525 patients, 39 (7%) were aged over 65. We compared 38 patients aged 16-64 years for comparison and reviewed the notes in detail. Elderly patients were mostly pedestrians (23/38 - 61%) though 21% (8/38) were drivers with 8% (3/38) on public transport. Younger patients were mostly drivers (14/37 - 38%), cyclists (9/37 - 24%) or motorcyclists (7/37 - 19%). Older patients had a higher median Injury Severity Score, p < 0.05, were more likely to be female (p<0.01), involved in RTA's between 9am-5pm (p<0.05) and have pre-existing medical conditions (p<0.01). The following were significantly increased in older patients (p values): surgical (<0.01), medical (<0.01) and therapist workload (p<0.05), complications (<0.0001), length of stay (<0.01). Less elderly were discharged directly to home (p<0.001). Strategies to reduce the mortality and morbidity associated with RTA in the elderly should emphasise a) older pedestrians during daytime hours, b) attention to safety in public transport. Treatment must ensure adequate medical and therapist input to anticipate higher complication rates. PMID- 11495233 TI - How informed is consent? Use of an information booklet in patients undergoing total hip replacement. AB - For informed consent, patients must understand the risks and benefits of the proposed procedure. Fifty patients undergoing total hip replacement (THR) participated in an evaluation of a newly devised information booklet. Patients' understanding of the risks and benefits of THR was assessed using a questionnaire before and after they had read the booklet. Pre-booklet patients knew on average 2 benefits of THR, compared with 3 afterwards. Twenty-two patients initially identified no alternative treatment to THR. Those who knew an alternative stated on average 1 treatment compared with 2 in the post-booklet group. Fifteen patients identified no risks of THR initially, compared with two after reading the booklet. Initially patients reported 1 risk, versus 3 after reading the booklet. Forty-eight patients thought that the booklet increased their understanding of THR and its risks. Fourteen found it upsetting, but acceptable. Forty-six patients wished to keep the booklet. PMID- 11495234 TI - Preventing accidental falls among older people in long stay units. AB - A fall prevention programme for older long-stay patients in a 95 bedded District Hospital was undertaken. Data on falls and resulting injuries for the year prior to the intervention were compared with equivalent data after one year (Year 1) and after two years (Year 2) of the intervention. In the pre-intervention year 25% of patients had at least one fall compared with 20.9% and 17.4% in Year 1 and Year 2 respectively. This difference was not statistically significant. However, there were 21% fewer falls in Year 1 and 49.3% fewer in Year 2 than in the pre intervention year. This difference was significant in Year 2. In both intervention years there was a significant reduction in the incidence of fracture from 20.5% of falls (pre-intervention) to 2.8% in Year 1 and no fractures occurred in Year 2. Significant reductions in soft tissue injuries occurred in Year 2 but not in Year 1, dropping from 38.5% (pre-intervention) to 36.1% and 15.4% respectively. The percentage of patients uninjured after a fall increased from 41% to 61.1% to 84.6%. This intervention reduced falls and their adverse consequences for older people living in the long stay unit. The effect of the intervention escalated in Year 2. The intervention cost IR4,800 pounds. Fall prevention should be part of the routine care of older people in all types of long stay care. PMID- 11495235 TI - A five year review of scar dehiscence in the Rotunda Hospital, Dublin. AB - The aims of this study were to identify risk factors for scar dehiscence in labour, to illustrate the clinical presentations of patients with scar dehiscence and to quantify the risk posed by the use of oxytocin in labour. This was a case controlled, 5 year retrospective study. Patients with scar dehiscence were identified from labour ward records with matched controls and chart review of case and control patients were performed. Our results showed that the vaginal delivery rate for trial of scar was 76.9%. The incidence of scar dehiscence was 0.043%. Oxytocic labour augmentation was a risk factor (OR 4.5, 95% CI 0.9313 42.8, p=0.065) but induction of labour using oxytocin was not (p=0.222). The commonest symptom of scar dehiscence was fetal distress (OR 12.3, 95% CI 1.9-81). There was no maternal or fetal mortality. We concluded that trial of labour after one caesarean section is acceptable practice with a good success rate and a low incidence of serious morbidity. The use of oxytocin to augment labour is associated with scar dehiscence. PMID- 11495236 TI - Folic acid and prevention of neural tube defects in 2000 improved awareness--low peri-conceptional uptake. AB - Eight years have passed since recommendations were made by the Irish Department of Health on the importance of folic acid in the prevention of neural tube defects (NTD). There is currently no mandatory fortification of foodstuffs with folic acid in Ireland, with reliance placed on campaigns promoting increased dietary folate intake and supplements. We assessed knowledge and use of folic acid among 300 women attending ante-natal clinics in Dublin maternity hospitals in the year 2000 using an interviewer administered questionnaire. Qualitative information was obtained through means of a focus group. Ninety two percent of respondents had heard of folic acid and 67% knew it could prevent NTD. Thirty per cent were advised to take it peri-conceptionally but overall only 18% did so; 39% of women had planned their pregnancy. The focus group indicated that folic acid was not 'visible' enough and that fortification of food was more realistic. This study shows that improved folic acid awareness has not been accompanied by corresponding peri-conceptional uptake in 2000. Folic acid promotional campaigns should be continuous and targeted. Mandatory food fortification should be strongly considered. PMID- 11495237 TI - Patent omphalomesenteric duct presenting as an umbilical discharge. PMID- 11495238 TI - A rare cause of intraperitoneal bleeding. PMID- 11495239 TI - Diagnostic review of 66 children with learning disability attending a single centre. PMID- 11495240 TI - Census 200--steps in the right direction, but still room for improvement. PMID- 11495241 TI - Ambulatory paediatrics: improving patient care. PMID- 11495242 TI - Structure of the N-terminal region of Haemophilus influenzae H10017: implications for function. AB - Haemophilus influenzae is a gram-negative pathogen that causes infections ranging from asymptomatic colonization of the human upper respiratory tract to serious invasive diseases such as meningitis. Although the genome of Haemophilus influenzae has been completely sequenced, the structure and function of many of these proteins are unknown. H10017 is one of these uncharacterized proteins. Here we describe the three-dimensional solution structure of the N-terminal portion of H10017 as determined by NMR spectroscopy. The structure consists of a five stranded antiparallel beta-sheet and two short alpha-helices. It is similar to the C-terminal domain of Diphtheria toxin repressor (DtxR). The C-terminal portion of H10017 has an amino acid sequence that closely resembles pyruvate formate-lyase--an enzyme that converts pyruvate and CoA into acetyl-CoA and formate by a radical mechanism. Based on structural and sequence comparisons, we propose that the C-terminus of H10017 functions as an enzyme with a glycyl radical mechanism, while the N-terminus participates in protein/protein interactions involving an activase (iron-sulfur protein) and/or the substrate. PMID- 11495243 TI - Quantitative measurement of water diffusion lifetimes at a protein/DNA interface by NMR. AB - Hydration site lifetimes of slowly diffusing water molecules at the protein/DNA interface of the vnd/NK-2 homeodomain DNA complex were determined using novel three-dimensional NMR techniques. The lifetimes were calculated using the ratios of ROE and NOE cross-relaxation rates between the water and the protein backbone and side chain amides. This calculation of the lifetimes is based on a model of the spectral density function of the water-protein interaction consisting of three timescales of motion: fast vibrational/rotational motion, diffusion into/out of the hydration site, and overall macromolecular tumbling. The lifetimes measured ranged from approximately 400 ps to more than 5 ns, and nearly all the slowly diffusing water molecules detected lie at the protein/DNA interface. A quantitative analysis of relayed water cross-relaxation indicated that even at very short mixing times. 5 ms for ROESY and 12 ms for NOESY, relay of magnetization can make a small but detectable contribution to the measured rates. The temperature dependences of the NOE rates were measured to help discriminate direct dipolar cross-relaxation from chemical exchange. Comparison with several X-ray structures of homeodomain/DNA complexes reveals a strong correspondence between water molecules in conserved locations and the slowly diffusing water molecules detected by NMR. A homology model based on the X-ray structures was created to visualize the conserved water molecules detected at the vnd/NK-2 homeodomain DNA interface. Two chains of water molecules are seen at the right and left sides of the major groove, adjacent to the third helix of the homeodomain. Two water-mediated hydrogen bond bridges spanning the protein/DNA interface are present in the model, one between the backbone of Phe8 and a DNA phosphate, and one between the side chain of Asn51 and a DNA phosphate. The hydrogen bond bridge between Asn51 and the DNA might be especially important since the DNA contact made by the invariant Asn51 residue, seen in all known homeodomain/DNA structures, is critical for binding affinity and specificity. PMID- 11495244 TI - A new approach for obtaining sequential assignment of large proteins. AB - A novel NMR experiment for obtaining sequential assignment of large proteins and protein complexes is described. The proposed method takes full advantage of transverse relaxation optimized spectroscopy (TROSY) and utilizes spin-state selection to distinguish between intraresidual and sequential connectivities in the HNCA-TROSY-type correlation experiment. Thus, the intra- and interresidual cross peaks can be identified without relaying magnetization via carbonyl carbon, which relaxes very rapidly at the high magnetic fields where TROSY is most efficient. In addition, the presented method enables measurement of several scalar and residual dipolar couplings, which can potentially be used for structure determination of large proteins. PMID- 11495245 TI - Improved 3D triple resonance experiments, HNN and HN(C)N, for HN and 15N sequential correlations in (13C, 15N) labeled proteins: application to unfolded proteins. AB - Two triple resonance experiments, HNN and HN(C)N, are presented which correlate HN and 15N resonances sequentially along the polypeptide chain of a doubly (13C, 15N) labeled protein. These incorporate several improvements over the previously published sequences for a similar purpose and have several novel features. The spectral characteristics enable direct identification of certain triplets of residues, which provide many starting points for the sequential assignment procedure. The experiments are sensitive and their utility has been demonstrated with a 22 kDa protein under unfolding conditions where most of the standard triple resonance experiments such as HNCA, CBCANH etc. have limited success because of poor amide, Calpha and Cbeta chemical shift dispersions. PMID- 11495247 TI - Novel 2D and 3D multiple-quantum bi-directional HCNCH experiments for the correlation of ribose and base protons/carbons in 13C/15N labeled RNA. AB - Multiple-quantum 2D and 3D bi-directional HCNCH experiments are presented for the correlation of base and ribose protons/carbons in 13C/15N labeled HIV-1 TAR RNA. In both 2D and 3D experiments, the magnetization of H1' is transferred to H6/H8 and H1' through H1'-C1'-N1/9-C6/8-H6/8 and H1'-C1'-N1/9-C1'-H1' pathways, and the magnetization of H6/8 is transferred to H1' and H6/8 through H6/8-C6/8-N1/9-C1' H1' and H6/8-C6/8-N1/9-C6/8-H6/8 pathways. Chemical shifts of four different nuclei (H1', C1', C6/8 and H6/8) are sampled in the 2D experiment. The correlation of base and ribose protons/carbons is established by the rectangular arrangement of crossover and out-and-back peaks in the proton/carbon correlated spectrum. The rectangular connections can be further resolved using the nitrogen dimension in a 1H/13C/15N 3D experiment. Furthermore, by taking advantage of the well separated chemical shifts of N1 (pyrimidine) and N9 (purine), the 2D spectrum can be simplified into two sub-spectra based on their base type. Both experiments were tested on a 13C/15N labeled 27-mer HIV-1 TAR RNA containing a UUCG hairpin loop. PMID- 11495248 TI - A TROSY relayed HCCH-COSY experiment for correlating adenine H2/H8 resonances in uniformly 13C-labeled RNA molecules. AB - A new TROSY relayed HCCH-COSY pulse sequence is introduced for correlating adenine H2 and H8 resonances in 13C-labeled RNA molecules. The pulse scheme provides substantial improvements in signal-to-noise compared to previously suggested experiments, and therefore will be suitable for NMR studies of larger RNA molecules. The experiment provides 13C chemical shifts for all carbon nuclei in the adenine base. This is advantageous for resolving spectral overlap in larger RNA molecules and provides a starting point for measuring additional parameters for these carbons in the adenine spin system. PMID- 11495246 TI - An improved method for distinguishing between anisotropic tumbling and chemical exchange in analysis of 15N relaxation parameters. AB - Although an accurate description of global tumbling of a protein is essential for correct analysis of internal motions. proper distinction between the effects of anisotropic rotational diffusion and conformational exchange has remained a challenge. We present a novel two-part filtering procedure designed specifically to distinguish between the effects of anisotropy and conformational exchange. The efficacy of this method is assessed using synthetic data sets. The method is then applied to two proteins of dramatically different size and shape, OspA and SH3. The large size and extreme anisotropy of OspA provide a challenging case, where conformational exchange is a small perturbation of the effects of anisotropy on transverse relaxation rates. Conversely, in the chicken c-Src SH3 domain, with its small size and nearly spherical shape, anisotropy is a small perturbation of the effects of conformational exchange on transverse relaxation rates. Accurate extraction of the global tumbling parameters for each protein allows optimal characterization of conformational exchange processes, as well as ps-ns time scale motions. PMID- 11495249 TI - TROSY NMR with partially deuterated proteins. AB - TROSY-type optimization of liquid-state NMR experiments is based on the preservation of unique coherence transfer pathways with distinct transverse relaxation properties. The broadband decoupling of the 1H spins interchanges the TROSY and anti-TROSY magnetization transfer pathways and thus is not used in TROSY-type triple resonance experiments or is replaced with narrowband selective decoupling. To achieve the full advantage of TROSY, the uniform deuteration of proteins is usually required. Here we propose a new and general method for 1H broadband decoupling in TROSY NMR, which does not compromise the relaxation optimization in the 15N-1H moieties, but uniformly and efficiently refocuses the 1JCH scalar coupling evolution in the 13C-1H moieties. Combined with the conventional 2H decoupling, this method enables obtaining high sensitivity TROSY type triple resonance spectra with partially deuterated or fully protonated 13C,15N labeled proteins. PMID- 11495250 TI - 1H, 15N, and 13C resonance assignments for the whole region 4 of Escherichia coli RNA polymerase sigma70 subunit. PMID- 11495251 TI - 1H, 13C and 15N chemical shift assignment of the honeybee pheromone carrier protein ASP1. PMID- 11495252 TI - Sequence-specific assignment and determination of the secondary structure of the 163-residue M. tuberculosis and M. bovis antigenic protein mpb70. PMID- 11495253 TI - Sequence-specific 1H, 13C and 15N signal assignments and secondary structure of archaeoglobusfulgidus SRP19. PMID- 11495254 TI - 1H, 15N and 13C resonance assignments and secondary structure of the liver ribonuclease from bullfrog Rana catesbeiana. PMID- 11495255 TI - 1H, 13C and 15N sequence-specific resonance assignment of the PSCD4 domain of diatom cell wall protein pleuralin-1 [etter]. PMID- 11495256 TI - Assignment of the 1H, 13C and 15N resonances of the PPIase domain of the trigger factor from Mycoplasma genitalium. PMID- 11495257 TI - The potential use of urinary excretion data for assessing the relative bioavailability of rifampicin in fixed dose combination anti-tuberculosis formulations. AB - SETTING: The perceived need for simple, non-invasive methods of assessing the relative bioavailability of rifampicin in fixed-dose combination (FDC) anti tuberculosis formulations. OBJECTIVE: To compare the performance of methods based on urinary excretion data with those utilising plasma concentration-time profiles to assess the relative bioavailability of rifampicin in combined and single-drug formulations. DESIGN: A two-period randomised crossover bioequivalence study in healthy male volunteers with a 1 week washout period between treatments. Plasma rifampicin concentrations were measured at 0, 1, 2, 4, 6 and 8 hours after each drug administration using a high performance liquid chromatography (HPLC) method. The rifampicin and desacetylrifampicin content of complete urinary collections made from 0-4 and 4-8 hours after dosage were determined using both the HPLC and a much simpler colorimetric procedure. RESULTS: There was good agreement between the relative bioavailability of the formulations using plasma and urinary excretion data, although the precision of the urinary-based estimates was slightly less than those derived from the plasma findings. There was also good agreement between the HPLC and colorimetric estimates of the combined urinary excretion of rifampicin plus desacetylrifampicin. CONCLUSIONS: Urinary excretion data may be used for ongoing quality control to confirm that commercial combined rifampicin-containing formulations that were initially shown to be satisfactory continue to be so. PMID- 11495258 TI - The influence of HIV infection and imprisonment on dissemination of Mycobacterium tuberculosis in a large Spanish city. AB - SETTING: Culture-positive cases of tuberculosis (TB) from the urban population of southern Madrid and from all the prisons located throughout the city. OBJECTIVE: To determine the frequency with which common strains of Mycobacterium tuberculosis cause disease among patients from both the urban and prison populations of a large Spanish city. DESIGN: Restriction fragment length polymorphism analysis was performed on culture-positive cases of TB identified between 1 January 1997 and 31 December 1998. Risk factors that might be associated with the dissemination of common strains of TB among the two populations were also investigated. RESULTS: Two hundred and twenty-one cases of culture-positive TB were identified, 99 (47.8%) of which were grouped in 23 clusters. Eleven were general clusters that spanned the prison and urban populations involved 69 patients (31.2%). Univariate analysis of risk factors showed that age <35 years, human immunodeficiency virus (HIV) infection, intravenous drug use and current or previous imprisonment were all associated at a statistically significant level with inclusion in general clusters. The final logistic regression model showed an interaction between HIV infection and incarceration. CONCLUSIONS: Dissemination of common strains of M. tuberculosis between prison inmates and the urban population of Madrid is significant, and involves subjects with a history of imprisonment and HIV infection. PMID- 11495259 TI - Tuberculosis in a prison population in Mwanza, Tanzania (1994-1997). AB - SETTING: Bugando Medical Centre, a referral consultant hospital in the lake zone to which all inmates from Butimba prison are admitted and treated. OBJECTIVE: To determine the extent of open (smear-positive) tuberculosis and other types of tuberculosis among prisoners with tuberculosis. DESIGN: A retrospective cohort study. Case notes of 501 prisoners from January 1994 to December 1997 were retrieved and reviewed. RESULTS: The proportion of open tuberculosis in this study was high, with 204 prisoners (40.7%) having smear-positive tuberculosis. Co infection was recorded in the majority of patients; HIV/AIDS was recorded in 25.9% of cases. The mean length of imprisonment at the time of diagnosis was 19 months. CONCLUSION: The proportion of open, smear-positive tuberculosis among prisoners admitted with tuberculosis is high. Intervention measures specifically targeting this population are urgently needed in order to contain tuberculosis disease in the prison population. PMID- 11495260 TI - The incidence of tuberculosis in drug users with small tuberculin reaction sizes. AB - SETTING: In persons infected with the human immunodeficiency virus (HIV), a decreased tuberculin reaction cut-point of > or = 5 mm induration is recommended. OBJECTIVE: To determine tuberculosis risk in non-anergic HIV-infected persons with 5-9 mm tuberculin reactions. DESIGN: A prospective study with semi-annual tuberculin and anergy testing, HIV antibody and T cell subset assays, and active surveillance for tuberculosis. RESULTS: Participants were 572 HIV-seronegative and 241 HIV-seropositive non-anergic drug users. No tuberculosis occurred in HIV seronegative persons. Tuberculosis incidence among HIV-seropositive drug users was 3.3, 7.7, 0, and 0.34 per 100 person-years in those with tuberculin reaction sizes of > or = 10 mm, 5-9 mm, 1-4 mm, and 0 mm, respectively, and was significantly increased in persons with 5-9 mm induration compared with those with 0-4 mm induration (rate ratio 27.7, 95%CI 2.9-268). Among persons with reaction sizes of 5-9 mm, tuberculosis occurred exclusively in those with CD4+ lymphocyte counts <500/mm3 at the time of their 5-9 mm tuberculin reactions. CONCLUSION: HIV-infected persons with tuberculin reaction sizes of 5-9 mm are at increased risk for tuberculosis compared to non-anergic persons with smaller (0-4 mm) reaction sizes. However, this increased risk may be limited to those with low CD4+ lymphocyte counts at the time of tuberculin testing. PMID- 11495261 TI - Screening for tuberculosis and latent tuberculosis infection among undocumented immigrants at an unspecialised health service unit. AB - SETTING: Practical or cost-effective strategies to identify undocumented immigrants with latent tuberculosis infection and to deliver treatment for latent TB infection are still unavailable. OBJECTIVES: To compare completion rates of screening procedures for TB infection and disease among undocumented immigrants at specialised (TB) and unspecialised health services in Italy. DESIGN: A TB unit (TBU) and an unspecialised health service unit for migrants (MHCU) served as recruitment sites for recent undocumented immigrants from TB endemic areas. The screening included a symptom questionnaire, a tuberculin skin test and a chest X ray. RESULTS: Of 1318 eligible subjects, 1232 (93.4%) accepted the screening. Screening was completed by 993 (80.6%) individuals overall, 86.5% and 71.4% at the TBU and MHCU services, respectively. In a multivariate analysis model, the only variable associated with an increased probability of completing screening was being enrolled at the TBU site (OR 2.5, 95%CI 1.8-3.5; P < 0.001). Three hundred and ninety-two subjects (39.4%) had a TST test of > or = 10 mm. Eight cases of active tuberculosis were detected, with a calculated prevalence of disease of 650/100,000. CONCLUSIONS: Undocumented immigrants to Italy can be screened for TB at an unspecialised health service unit, although not as efficiently as at a specialised TB unit. PMID- 11495262 TI - Efficacy of the BCG revaccination programme in a cohort given BCG vaccination at birth in Hong Kong. AB - SETTING: Revaccination of tuberculin-negative school-children is a regular practice in Hong Kong. OBJECTIVE: To assess the efficacy of BCG revaccination guided by tuberculin skin testing. METHOD: A cohort of 303,692 children vaccinated at birth was followed through the tuberculosis notification register for the development of active disease. The percentage of cohort who participated in the BCG revaccination program during primary school was estimated from the vaccination statistics of the Hong Kong Department of Health. The BCG revaccination history of identified cases was ascertained through vaccination cards and clinic records. RESULTS: A total of 85.2% of the cohort participated in the BCG revaccination programme; 79.7% of the participants were tuberculin negative and revaccinated; 343 developed tuberculosis after the age of 11; 302 were among the participants in the programme while 41 were not. The tuberculosis incidence was 16.5 and 12.9 per 100,000 person-years for participants and non participants, respectively (RR 1.28, 95%CI 0.92-1.77). Among the participants, tuberculosis incidence was 12.5 and 32.0/100,000 person-years, respectively, for the tuberculin-negative/BCG revaccinated group and the tuberculin-positive/non revaccinated group (RR 0.39, 95%CI 0.31-0.49). CONCLUSION: This study failed to demonstrate any significant difference in the incidence rates of tuberculosis among participants and non-participants in a school BCG revaccination programme. The increased risk for tuberculosis in the tuberculin-positive group does not support the use of the tuberculin testing for detection of immunity conferred by neonatal BCG vaccination. PMID- 11495263 TI - Molecular and conventional epidemiology of tuberculosis in an inner city district. AB - SETTING: Molecular epidemiology has underlined the importance of recent tuberculosis (TB) transmission and has uncovered notable discrepancies compared with conventional epidemiology. OBJECTIVES: 1) To determine, by RFLP analysis, the percentage of clustered cases in an inner city district with a high incidence of TB (163/100,000) and the groups at risk of being clustered; and 2) to compare the role of conventional contact tracing (CCT) with that of RFLP. DESIGN: RFLP was carried out using the IS6110 and pTBN12 (PGRS) sequences of 165 cultures positive for Mycobacterium tuberculosis in the Ciutat Vella district of Barcelona during 1997-1998. Contact tracing was carried out in 171 of 251 declared cases (68.1%). Associations were assessed by calculating odds ratios (OR) with 95% confidence intervals (CI). Logistic regression was used for multivariate analysis. RESULTS: Using RFLP, 76 (46.0%) strains were found to be clustered by IS6110 and PGRS. From CCT, 30 new patients were found among 858 contacts (3.5%) and 57 patients were linked. In terms of RFLP and CCT, the main risk factor was intravenous drug use (IVDU). In 44 cases who lived alone and were not involved in CCT, 50% were in RFLP clusters. The concordance rate between RFLP and CCT was 8/13 (61.5%); the disagreement corresponded to a cluster of five recent immigrants from Africa. Subsequent to RFLP, an epidemiological connection was found in 15/55 cases (27.2%). CONCLUSIONS: The percentage of clustered cases is very high. CCT was useful for identifying new cases, but it was insufficient for detecting the pathways of transmission. CCT coverage needs to be improved in marginalized individuals, and the results correlated with those of RFLP. PMID- 11495264 TI - DOT for patients with limited access to health care facilities in a hill district of eastern Nepal. AB - SETTING: The hill district in Nepal, where access to health care facilities is difficult. OBJECTIVE: To compare results before and after a decentralised directly observed treatment (DOT) intervention. DESIGN: Prospective study of patients registered in Dhankuta district, Nepal, 1996-1999. Patients received their intensive phase treatment under health worker supervision via one of three DOT options: 1) ambulatory from the peripheral government health facilities; 2) ambulatory from an international non-governmental organisation (INGO) TB clinic in district centre; or 3) resident in INGO TB hostel in district centre. Historical data from 1995-1996, with unsupervised short-course chemotherapy, were used for comparison. RESULTS: Of 307 new cases, respectively 126 (41%), 86 (28%) and 95 (31%) took their intensive phase treatment via options 1, 2 and 3. Smear conversion (at 2 months) and cure rates in new smear-positive pulmonary tuberculosis cases were respectively 81.6% (vs. 58.8% historical, P = 0.001) and 84.9% (vs. 76.7% historical, P = 0.03). Overall costs to the INGO provider fell by 7%, mainly as a result of staffing reductions in the INGO services made possible by rationalisation with government services during the intervention. CONCLUSION: By offering varied DOT delivery routes, including an in-patient option, satisfactory results are possible with DOT even in areas where access to health care facilities is difficult. Provision of in-patient care via an INGO TB hostel allowed a significant proportion of new cases (31%) to receive their intensive phase treatment who otherwise may have had difficulty accessing treatment, due either to the distance to the nearest health facility or to disease severity. Substitution of government hospital beds or local hotel beds for the INGO hostel beds may allow the model to be reproduced elsewhere in similar geographical conditions in Nepal, but further studies should be performed in a non-INGO supported district beforehand. PMID- 11495265 TI - Feasibility of home-based and health centre-based DOT: perspectives of TB care providers and clients in an HIV-endemic area of Thailand. AB - Focus groups were conducted in a high human immunodeficiency virus (HIV) prevalence area of Thailand to elicit perspectives of health staff and clients regarding the feasibility of directly observed therapy (DOT) for tuberculosis. Most participants perceived health centre-based DOT to be impractical for clients due to severe illness, travel inconvenience, and interference with employment. Most providers perceived home-based DOT to be difficult because of the inconvenience of travel, staff shortages and the high tuberculosis caseload. Most clients except HIV-negative tuberculous females considered home visits to be undesirable due to stigma. The preparedness of providers for home-based DOT might be improved through awareness building among staff about multidrug-resistant tuberculosis. PMID- 11495266 TI - Evaluation of suspected tuberculous pleurisy: clinical and diagnostic findings in HIV-1-positive and HIV-negative adults in Uganda. AB - SETTING: National Tuberculosis Treatment Centre, Mulago Hospital, Kampala, Uganda. OBJECTIVES: To compare clinical and radiographic presentation, and diagnostic methods, in adults with tuberculous pleurisy who are negative and positive for the human immunodeficiency virus (HIV). DESIGN: Adults with suspected pleural tuberculosis were screened by clinical examination, thoracocentesis and closed pleural biopsy. Biopsy material was cultured on Middlebrook 7H-10 solid medium and in BACTEC 12B radiometric vials. Pleural fluid was cultured using Lowenstein-Jensen slants, BACTEC and Kirchner liquid medium. RESULTS: Of 156 individuals enrolled, 142 had tuberculosis, of whom 80% were HIV positive. Among those with tuberculosis, HIV-positive patients bad a more severe and longer illness. The size of effusions was similar in HIV-positive and HIV negative patients. A higher proportion of HIV-positive patients had parenchymal infiltrates but this difference was not statistically significant. Pleural fluid lymphocytosis was present in all HIV-negative and 97% of the HIV-positive patients. HIV-positive patients had lower pleural fluid lymphocyte counts. Pleural fluid cultures were more often positive in HIV-positive patients. BACTEC and Kirchner liquid media gave higher yields than solid media. CONCLUSION: HIV positive patients with tuberculous pleurisy had a more severe illness than HIV negative patients. Mycobacterial cultures from HIV-positive patients were more often positive, suggesting more mycobacterial extension from the lungs into the pleural space. Liquid culture media were superior to solid media with regard to diagnostic yield and time until diagnosis. PMID- 11495267 TI - Diagnostic value of a nested polymerase chain reaction assay on peripheral blood mononuclear cells from patients with pulmonary and extra-pulmonary tuberculosis. AB - SETTING: Saint Louis Hospital, Paris, France. OBJECTIVE: To determine the clinical relevance of detection of Mycobacterium tuberculosis DNA by nested polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMCs) in the rapid diagnosis of tuberculosis. DESIGN: Single-centre prospective case study of 90 hospitalised patients and 50 healthy subjects or blood donors from 1 January to 30 June 1998. RESULTS: Twenty-three patients were diagnosed with tuberculosis (26.7%); 20 tuberculosis patients were culture-positive, with seven smear-positive for acid-fast bacilli. Sensitivity of smear, culture and nested PCR was 30.4 (7/23), 87 (20/23) and 30.4% (7/23), respectively. The specificity of smear and culture was 100%, and the specificity of the nested PCR was 96% in the healthy subjects. However, the specificity decreased to 83.6% in the hospitalised patients, with 11 nested PCR-positive patients without a diagnosis of tuberculosis. The sensitivity of the nested PCR was low in pulmonary tuberculosis (22.2%), but increased in pulmonary/extra-pulmonary tuberculosis (50%), extra-pulmonary tuberculosis (33%), and disseminated tuberculosis (33%). CONCLUSION: The use of a nested PCR assay on PBMC may pose problems for the rapid diagnosis of tuberculosis with regard to low sensitivity and specificity. However, further studies are needed to confirm this technique as an alternative test for the diagnosis of paucibacillary forms of tuberculosis. PMID- 11495268 TI - Mycobacterium tuberculosis bacteremia in a university hospital. AB - SETTING: Patients with blood cultures positive for Mycobacterium tuberculosis between 1988 and 1999. OBJECTIVE: To study the clinical and microbiological characteristics of patients with tuberculous bacteremia, including data about evolution and management. DESIGN: Retrospective review of the clinical charts and microbiological records of patients with culture-proven tuberculous bacteremia between 1988-1999. RESULTS: During the study period, 19 patients with culture proven M. tuberculosis bacteremia were detected (1.42 isolates/patient, 4.7% of all patients with blood cultures for mycobacteria). Four patients were non infected with the human immunodeficiency virus and 15 were HIV-infected. In four patients blood was the only positive sample. Five patients were diagnosed simultaneously with tuberculosis and HIV infection. Only 13 had a temperature higher than 37.5 degrees C. Most patients had symptoms or signs of respiratory tract involvement, and 11 patients died (10 from tuberculosis). The average time for detection of positive blood cultures was 33.25 days for lysis-centrifugation cultures and 26.46 days for BACTEC cultures. The incidence of M. tuberculosis bacteremia remained stable during the study period. CONCLUSIONS: Although blood cultures are useful for definitive diagnosis of disseminated tuberculosis, the long incubation times made them of limited usefulness in the clinical management of patients. Mortality remains high in these patients. PMID- 11495269 TI - Treatment of ulcerative endobronchial tuberculosis and bronchial stenosis with aerosolized streptomycin and steroids. AB - OBJECTIVES: Endobronchial tuberculosis (EBTB) is defined as a tuberculous infection of the tracheobronchial tree. It has been reported that aerosolized therapy with streptomycin and steroids is useful for EBTB; however, the effectiveness of this therapy for bronchial stenosis has yet to be clarified. This study was undertaken to determine the effectiveness of aerosol therapy in the treatment of bronchial stenosis due to EBTB. DESIGN: An observational, historical, controlled comparative study. Retrospective analysis of 27 patients treated with conventional therapy, and prospective analysis of 30 patients treated with aerosol therapy. METHOD AND PATIENTS: Flexible bronchoscopy was performed at least twice in 57 patients with ulcerative EBTB, in whom the degree of bronchial stenosis between the first and last bronchoscopic examinations was estimated. Bronchial stenosis was graded as minimal, mild, moderate, severe or obstructive, and the follow-up of bronchial stenosis assessed as aggravation, no change or improvement. RESULTS: Conventional therapies led to aggravation in 13 patients, no change in 13 patients, and improvement in one patient. Aerosol therapy led to no change in 27 patients, and improvement in three patients. No patients developed aggravation. The differences between the therapeutic groups were significant. CONCLUSION: Aerosol therapy helps to prevent progressively severe bronchial stenosis due to EBTB. PMID- 11495270 TI - The epidemiology of adult lung disease in Botswana. AB - SETTING: Republic of Botswana. OBJECTIVES: To describe and analyse the epidemiology of adult lung disease in Botswana and provide information about risk factors. DESIGN: Register-based retrospective study. RESULTS: Poverty and human immunodeficiency virus (HIV) infection are major risk factors for lung disease. In 1997, TB caused 15.7%, pneumonia 8.3%, asthma/COPD 0.7% and lung cancer 0.4% of adult in-patient deaths, while 15.5% of deaths were classified as AIDS. Respiratory conditions accounted for 14.2% of hospital admissions and 18.1% of out-patient consultations. Pneumoconiosis was under-reported. Classification problems exist between AIDS, TB and pneumonia. The case fatality rate for in patients with pneumonia was 5.6% in 1990 vs. 15.5% in 1997, and for TB patients it was 7.4% in 1990 vs. 11.2% in 1997. Morbidity rates also increased for pneumonia and TB through the 1990s. There was a steeper rise in pneumonia mortality and morbidity rates in females than males. The gender distribution in TB incidence remained unchanged, but the average age of female TB patients decreased by 7.7 years from 1983 to 1998, vs. a decrease of 5.3 years in males. CONCLUSIONS: Pulmonary TB and pneumonia are major causes of adult mortality and morbidity in Botswana. Incidences and case fatality rates are increasing, due to co-existent HIV infection. Non-communicable lung diseases are less common causes of death, and prevalences seem to be stable. PMID- 11495271 TI - The potency of tuberculin PPD RT23. PMID- 11495272 TI - Disseminated Mycobacterium tuberculosis infection of the upper respiratory tract diagnosed by spoligotyping. PMID- 11495273 TI - Resection of the thoracic vertebral bodies to relieve the pressure on the major airways: an approach to reduce respiratory symptoms due to tuberculosis sequelae. PMID- 11495274 TI - Using other people's graphs. PMID- 11495275 TI - Radioprotective, anticarcinogenic and antioxidant properties of the Indian holy basil, Ocimum sanctum (Tulasi). PMID- 11495276 TI - Germins and germin like proteins: an overview. AB - Molecular investigations during wheat germination have revealed unique developmentally regulated proteins, designated as germins, which show remarkable resistance to broad specificity proteases and to dissociation in SDS. Germins in cereals have an oxalate oxidase activity, which generates H2O2 from the oxidative breakdown of oxalate thereby playing a significant role in plant development and defense. Germin like proteins (GLPs) exhibit sequence and structural similarity with the cereal germins but mostly lack oxalate oxidase activity. Germins and germin like proteins (GLPs) are a class of developmentally regulated glycoproteins characterized by a beta-barrel core structure, a signal peptide, and are associated with the cell wall. GLPs exhibit a broad range of diversity in their occurrence and activity in organisms ranging from myxomycetes, bryophytes, pteridophytes, gymnosperms and angiosperms. Germins and GLPs are thought to play a significant role during zygotic and somatic embryogenesis (wheat and Pinus, respectively), salt stress (barley and Mesembryanthemum crystallinum), pathogen elicitation (wheat and barley), and heavy metal stress, etc. Characterization and cloning of some of the genes encoding germins and GLPs has facilitated a better understanding of their regulation and raised their potential of biotechnological application. PMID- 11495277 TI - Liposome encapsulated tumor-associated antigens elicited humoral and cellular immune responses in mice bearing tumor. AB - Chemically induced tumors in mice provide a system to investigate tumor associated antigens (TAA). The cell surface glycoprotein antigens on such tumor cells have been identified as suitable targets for immune attack. The induction of immune responses against (TAA) in N-nitrosodiethylamine (DEN) exposed mice has been examined. In order to present antigens to the immune system, the liposome was used as vehicle to deliver the TAA. Liposomal-TAA formulation, elicited both humoral and the cellular immune responses, when administered intramuscularly in DEN-exposed mice. Presence of circulatory antibodies against TAA and the induction of cellular responses in immunized mice were monitored using ELISA and in vitro cell proliferation assay of lymphocytes respectively. Specificity of antibody against TAA in immune sera was analysed using immunoblotting technique. Based on these results, it is proposed that the liposome encapsulated TAA may successfully be used to induce humoral and cellular immune responses against tumor. PMID- 11495278 TI - Immunochemical and functional characterization of a polyclonal antibody to human sperm antigen. AB - A polyclonal antibody was raised against a 16 kDa human sperm protein identified by a monoclonal antibody to human sperm. The antibody showed significant reactivity with mouse spermatozoa as seen by ELISA. Immunohistochemical analysis showed that the antibody reacted with antigens from mouse testis, prostate as well as seminal vesicle. In both mouse and human testis the antibody localized antigens in round as well as elongated spermatids and mature spermatozoa. By SDS PAGE and Western blot analysis the antibody reacted with a 16 kDa protein in the testis and seminal vesicle, whereas in the prostate it identified two proteins, one at 20 kDa and another at 25 kDa. Immunofluorescent localization by the antibody showed reactivity with acrosomal and/equatorial and midpiece region of human spermatozoa. The antibody showed extensive agglutination both in mouse and human spermatozoa. The results indicate that the antigen may be a conserved antigen. Cross reactivity of the antibody with mouse spermatozoa enabled us to carry out antifertility trials. Passive immunization of female mice with this antibody caused 67% reduction in fertility. It is likely that the antifertility effect could be partly due to agglutinating nature of the antibody which may have caused inhibition of all processes that depend on forward motility such as cervical mucus penetration and possibly preventing sperm egg interaction. Such well characterized and functionally relevant antibodies will enable to identify sperm antigens relevant for fertility. Identification of such antigens may also help in diagnosis of immuno infertility. PMID- 11495279 TI - Apoptosis in endometrium of mouse during estrous cycle. AB - The present study was carried out to evaluate apoptosis in endometrium and to correlate these changes with the circulating levels of estradiol and progesterone in the mouse. Apoptosis was observed in various compartments of mouse uterus i.e. stroma, glandular epithelium and luminal epithelium depending on the stage of cycle. Stromal cell apoptosis was observed during various stages of cyclicity except on estrus day. Luminal epithelial cells showed apoptotic changes during all stages of cyclicity except on diestrus day. During metestrus, apoptosis was observed in glandular and luminal epithelia as well as stromal cells. Steroid antagonists such as tamoxifen and onapristone altered the apoptotic changes in the uterus. The results suggest that epithelial cell apoptosis is regulated by estrogen while stromal cell apoptosis is under the control of progesterone. PMID- 11495280 TI - Effect of single prenatal haloperidol exposure on hippocampus and striatum of developing rat brain. AB - Poor development and differentiation of three layered cytoarchitectural pattern of brain, degenerating pyramidal cells with pyknotic nuclei and substantial loss of both large and small pyramidal cells of the hippocampal CA1 region were observed in fetuses of pregnant Charles-Foster rats exposed to single high dose of haloperidol (50 mg/kg body weight) on day 12 of gestation. In treated striatum, reduction in size, complete degeneration of multipolar cells with fragmented nuclei and increased extracellular spaces were observed. Unsacrificed group of day 12 haloperidol treated rat offsprings at 9 weeks of age exhibited cognitive behavioural dysfunctions in passive avoidance (behaviour) test. These findings indicate that a single (high dose) prenatal haloperidol exposure during critical period of CNS development not only induces micromorphological aberrations in foetal hippocampus and striatum but also lasting cognitive impairment in adult rat offsprings. PMID- 11495281 TI - Effect of short photoperiod on organ growth kinetics and serum hormone profile in pullets of domestic fowl, Gallus gallus domesticus. AB - Effect of short photoperiod (SP; LD 6:18) treatment on serum hormone profile and growth rate of adrenal, thyroid, ovary, oviduct, liver and lymphoid organ was studied in rearing pullets (RIR breed) of 1 to day 90 old. Body weight and growth index of SP pullets were lesser as compared to pullets reared under LD 12:12. Except for ovary (recorded marginal increment), weights and growth indices of thyroid, adrenal and oviduct decreased under SP. Weight of liver and lymphoid organs was higher at 30 and 90 days, in SP pullets as compared to LD 12:12. Histometric data suggested that the transition from small to big follicles was slow in ovary of SP pullets, and also reduced follicular atresia was noted in SP pullets. Except for higher corticosterone level at 30 days and higher progesterone level at 30 and 60 days, relative levels of all the hormones at all other ages were lower in SP pullets. In general, the present observations suggested intraovarian changes in pullets exposed to SP. PMID- 11495283 TI - Butyrate induced alterations during development of the strain ln(1)BM2(reinverted) of Drosophila melanogaster. AB - The polytene male X chromosome of D. melanogaster, has a unique morphology, which is correlated with the property of increased transcription of the sex-linked genes of the male X chromosome. This ensures equalization of X-linked gene products between males (XY) and females (XX). Till date, an invariable correlation between the structure and transcription of the male X chromosome has been reported. However, the strain In(1)BM2(reinverted) of D. melanogaster presents a caveat to this invariable correlation. In this strain, although the male X chromosome appears puffy and diffuse, the transcription remains at the wild type level. This observation suggests the perturbation in the function of a regulator that controls the structure of the male X chromosome. In this report the response of the strain to butyrate, an inhibitor of histone deacetylase, has been studied, with specific reference to development, sex ratio and chromosome morphology of the strain. Two important conclusions arise from these experiments : (a) exposure to butyrate has more severe consequences on the development of the mutant strain and on the survival of females. (b) rearing on butyrate induced a temporal series of structural alteration of the polytene chromosome of the wild type, with the male X chromosome being most vulnerable to structural changes. The butyrate-interaction of In(1)BM2(reinverted) together with our current biochemical analyses of a chromosome coiling protein recovered from this strain, provide data for a working hypothesis explaining the sex and chromosome specific alteration of the structure of the male X chromosome of In(1)BM2(reinverted). PMID- 11495282 TI - Biochemical mechanism of combined effect of ethambutol and sparfloxacin against Mycobacterium smegmatis. AB - M. smegmatis cells grown in the presence of combination of ethambutol (EMB) and sparfloxacin (SPX) had decreased level of total cellular lipids as compared to control as well as cells grown in the presence of sub-inhibitory concentration (MIC50) of individual drugs. Amongst various phospholipids analyzed, maximum decrease was observed in the content of phosphatidylinositolmannosides (PIMs) of the cells grown in combination of EMB and SPX. In contrast, the subcellular distribution of phospholipids revealed a significant increase in PIMs content of both cell wall and cell membrane of the cells grown in the presence of combination of drugs as compared to control as well as individual drugs. Mycolic acids of M. smegmatis cells were found to be main targets as combination of drugs resulted in significant decrease in total cellular as well as cell wall mycolic acids as compared to control and individual drugs. Changed lipid composition of M. smegmatis cells grown in the presence of MIC50 of EMB, SPX and combination resulted in significant surface changes as was evident from decreased limiting fluorescence (Fmax) intensity of 1-anilinonaphthalene-8-sulfonate (ANS). Thus, the results of this study suggested that ethambutol and sparfloxacin in combination exerted their antimycobacterial effect principally due to their action on phosphatidylinositolmannosides (PIMs) and mycolic acids, which form the permeability barrier of mycobacteria. PMID- 11495284 TI - Studies on stibanate resistant Leishmania donovani isolates of Indian origin. AB - Studies with 26 clones of L. donovani promastigotes derived from three different Indian isolates indicated that wild type parasites are mixture of stibanate sensitive and resistant cells. Both forms of the parasite were resistant to the drug. Infection with resistant parasites appears to be the primary reason of high rate of pentavalent antimony unresponsiveness among Indian kala-azar patients. It was observed that the resistant parasites originated as a result of irregular and often incomplete treatment of kala-azar patients with pentavalent antimonials. PMID- 11495285 TI - Identification of a 75 kDa highly immunodominant antigen from Mycobacterium smegmatis and cross-reactivity with other species. AB - Three monospecific antibodies MSAb 1, MSAb 2 and MSAb 3 were raised in BALB/C mice against respective antigens. M. smegmatis whole cell lysate was first separated on SDS-PAGE and randomly chosen bands were cut and then used for immunization. Antibodies were collected as ascites by injecting mice with myeloma cell line P3X63 Ag 658.4. All the three antibodies showed high reactivity with denatured antigens compared to native. Different extent of cross-reactivity was observed as evident from ELISA. MSAb1 recognized a 75 kDa immunodominant antigen from M. smegmatis and 66 kDa from M. tuberculosis (H37Ra), respectively. An apparently similar molecular weight antigen shown to be present in M. tuberculosis (H37Ra) an avirulent strain and BCG, but not recognized by MSAb1. The 75 kDa antigen has a stimulatory effect on T-cell proliferation. PMID- 11495286 TI - Molluscicidal activity of Lawsonia inermis and its binary and tertiary combinations with other plant derived molluscicides. AB - Molluscicidal activity of leaf, bark and seed of Lawsonia inermis against Lymnaea acuminata and Indoplanorbis exustus was studied. Highest toxicity was observed in the seed of Lawsonia inermis. Toxicity of binary (1:1) and tertiary (1:1:1) combinations of the essential oil of cedar (Cedrus deodara Roxh) and neem (Azadirachta indica A. Juss), powder from bulb of garlic (Allium sativum Linn), and oleoresin extracted from rhizome of ginger (Zingiber officinale Rosc) with Lawsonia inermis and Embelia ribes fruit powder were studied against L. acuminata and I. exustus. L. inermis seed powder in combination with Cedrus deodara oil and Azadirachta indica oil was more toxic than their individual components and other combinations. PMID- 11495287 TI - Effect of dietary protein on zinc phosphide toxicity in albino rats (Rattus norvegicus). AB - Adult male and female albino rats (R. norvegicus) were administered rodenticide, zinc phosphide ranging from 4, 2, 1, 0.5, 0.25 and 0.125% in the diet containing 10(A), 17(B) and 24%(C) protein. Zinc phosphide induced 100% mortality at 4, 2, 1, 0.5% in diet A; 4 and 2% in diet B; and 4% in diet C. The results reveal influence of dietary protein in modulating the toxicity of zinc phosphide, suggesting that greater caution should be exercised while formulating its baits for effectiveness. The results also suggest that baits having 10% protein are more suitable to carry the rodenticide from view point of acceptability and efficacy towards the target species. PMID- 11495288 TI - Influence of chronic treatment of rats with isoprenaline and calcium channel blockers on response of isolated right ventricle to noradrenaline. AB - Influence of chronic treatment of rats with and calcium channel blockers (CCBs) and isoprenaline (ISP) on responses to noradrenaline (NA) was investigated on electrically--driven isolated right ventricle preparations. The ventricles were obtained from animals treated with chronic ISP or CCBs alone and chronic nifedipine, verapamil, diltiazem or nimodipine plus chronic ISP. A decreased response to NA as evidenced by an increase in EC50 for contraction which was observed in chronic ISP- treated preparations may be due to development of desensitisation (down-regulation) of beta-adrenoceptors. In chronic CCB-treated preparations there was a significant decrease in the EC50 of NA and decreased contractile response suggesting an increase in the beta-adrenoceptors and decreased availability of calcium, respectively. In chronic CCBs + ISP treated preparations further decreases in the EC50 values were observed suggesting that the voltage gated L-type Ca2+ channels may be affected directly or indirectly by change in beta-adrenoceptor activity. By the above results a proposed mechanism of interrelationship of beta-adrenoceptors with voltage gated L-type calcium channels in cardiac muscle is supported. PMID- 11495289 TI - Methyl parathion induced alterations in monoaminergic system of developing rat pups. AB - Methyl parathion induced alterations in the level of monoamines, viz. norepinephrine, dopamine and serotonin were studied in discrete regions of developing central nervous system of rat pups. A significant decrease in the level of monoamines noticed in methyl parathion toxicosis may be related to the altered neuronal activity and inefficiency, leading to depression and impairment in various behavioural activities. In contrast to AChE inhibition, monoamine oxidase (MAO) activity showed an increasing trend and it could cause deamination of catecholamines and accumulation of its metabolites. This suggests that an increased AChE inhibition may indirectly stimulate MAO activity in developing rat pups exposed to methyl parathion. PMID- 11495290 TI - Alkyl ester prodrugs for improved topical delivery of ibuprofen. AB - Topical delivery of ibuprofen directly to the site of inflammation can overcome gastrointestinal side effects associated with its long term oral administration. The set of physicochemical properties necessary for optimum topical delivery of ibuprofen can be imparted by formation of its ester prodrugs. Various alkyl ester prodrugs (methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-buty, n-pentyl, hexyl, heptyl, octyl, lauryl, cetyl and octadecyl esters) were synthesised and studied for their physicochemical properties and activity in the carrageenan induced rat paw oedema by topical route. Favourable shift in lipophilicity and self penetration enhancing effect of prodrugs responded in improved topical activity over the parent drug ibuprofen. PMID- 11495291 TI - Hypoglycemic effect of Hibiscus rosa sinensis L. leaf extract in glucose and streptozotocin induced hyperglycemic rats. AB - Investigations were carried out to evaluate the effect of aqueous extract of H. rosa sinensis leaves on blood glucose level and glucose tolerance using Wistar rats. Repeated administration of the extract (once a day for seven consecutive days), at an oral dose equivalent to 250 mg kg(-1), significantly improved glucose tolerance in rats. The peak blood glucose level was obtained at 30 min of glucose load (2 g kg(-1)), thereafter a decreasing trend was recorded up to 120 min. The data exhibit that repeated ingestion of the reference drug tolbutamide, a sulphonylurea and the extract brings about 2-3 fold decrease in blood glucose concentration as compared to single oral treatment. The results clearly indicate that tolbutamide improves the glucose tolerance by 91% and extract does so only by 47%. At 250 mg kg(-1), the efficacy of the extract was 51.5% of tolbutamide (100mg kg(-1)). In streptozotocin diabetic rats, no significant effect was observed with the extract, while glibenclamide significantly lowered the glucose level up to 7 hr. These data suggest that hypoglycemic activity of H. rosa sinensis leaf extract is comparable to tolbutamide and not to glibenclamide treatment. PMID- 11495292 TI - Midgut specific immune response of vector mosquito Anopheles stephensi to malaria parasite Plasmodium. AB - Innate immune-related polypeptides expression in midgut in the ageing vector mosquito A. stephensi following infection by malaria parasite, Plasmodium yoelii yoelii has been studied. Twenty polypeptides were induced by an infected blood meal during various stages of adult life. A 24 kDa polypeptide was induced generally in most of the stages. Maximum parasite induced polypeptides i.e. 22, 33, 111, 122, 127, 140, 143 and 146 kDa were found in 5 days of post blood feeding (PBF) which coincides with the presence of oocysts on the midgut. However, in addition, three polypeptides in 11 days PBF and 8 polypeptides in 20 days PBF were also induced due to parasite infection in aged mosquitoes. Quantitatively, the amount of soluble proteins in the midgut in oocyst-sporozoite positive mosquitoes was always less as compared to their normal counterparts. The parasite evidently elicits defined immune responses by inducing specific polypeptides in the midgut of the mosquito. PMID- 11495293 TI - Correlation of total and specific serum immunoglobulin E levels with the severity of chronic sinusitis in children. AB - Chronic sinusitis is frequently associated with allergy and asthma. Previous studies have shown that serum immunoglobulin E (IgE) levels correlate with allergy and asthma in adults. Because the role of allergic inflammation in the severity of chronic sinusitis remains controversial in children, we set out to determine whether a correlation exists between serum levels of total and specific IgE and the severity of chronic sinusitis in children. Forty-four children with chronic sinusitis were enrolled in the study. Computed tomographic scans were reviewed and scored for the severity of sinusitis. All children were mite sensitive. Serum samples were assayed for total IgE and specific IgE antibodies to mite allergen using a fluoroimmunoassay. Fourteen subjects had extensive sinus disease. There was no significant difference in the average of total and specific IgE between the subjects with extensive and limited disease (p = 0.562 and 0.755, respectively). Thirty-four subjects were diagnosed with asthma. The subjects with extensive sinus disease had a higher prevalence of moderate to severe asthma than the subjects with limited disease (p = 0.006), but there was no significant difference in the average of total and specific IgE between the subjects with different severities of asthma. (p = 0.833 and 0.425, respectively). The data suggests that levels of total or specific IgE do not correlate with severity of chronic sinusitis in children. Nonetheless, the severity of chronic sinusitis and asthma correlate well with each other irrespective of total and specific IgE. PMID- 11495294 TI - Low dose intravenous immunoglobulin for acute immune thrombocytopenic purpura in children. AB - Acute immune thrombocytopenic purpura (ITP) is a self-limited autoimmune disorder to platelets. The disease responds well to intravenous immunoglobulin (IVIG) treatment. We studied the efficacy of low dose (1 g/kg) IVIG to treat acute ITP in children. Of 17 children with ITP and platelet counts < 20,000/microl, 13 (76%) had rapid platelet count recovery, reaching safe level (> 50,000/microl) within 4 days after 1 g/kg IVIG. Four children needed a second dose. In 5 of 15 patients, platelet counts recovered to normal without recurrence, while in 10 patients, platelet counts declined again 2-3 weeks after the initial treatment; 6 of whom (40%) needed re-treatment. All but one patient had complete recovery of the platelet count by 6 months. Adverse effects of low dose IVIG were minimal. We recommend that for childhood ITP, 1 g/kg IVIG should be tried initially. If inadequate response is seen (platelet count < 30,000/microl) by 48 hours, a second dose is needed. PMID- 11495295 TI - Cytotoxic T lymphocyte responses to vaccinia virus antigens but not HIV-1 subtype E envelope protein seen in HIV-1 seronegative Thais. AB - The HIV-1 prime boost phase I/II vaccine trial using a recombinant canarypox vector, vCP1521, containing subtype E env (gp120), and subtype B env (gp41), gag and protease has started in Thailand. We have demonstrated that although 4 from 15 human immunodeficiency virus type 1 (HIV-1) seronegative Individuals showed cytotoxic T lymphocyte (CTL) responses to vaccinia virus antigens, none of them showed specific CTL responses to subtype E Env after in vitro stimulation. This preliminary study suggests that specific CTL responses to subtype E envelope detected in HIV-1 seronegative Individuals after vaccination should be considered as specific responses to the immunization. PMID- 11495296 TI - Use of complement receptor 1 (CD35) assay in the diagnosis and prognosis of immune complex mediated glomerulopathies. AB - Complement Receptor 1 (CR1) is a polymorphic glycoprotein expressed on erythrocytes, leukocytes and glomerular podocytes and has a major role in immune complex processing. In addition, it regulates the complement cascade activation by preventing formation of classical and alternative pathway convertases and by acting as a cofactor for Factor I mediated cleavage of C3. In this study, we have examined the expression of erythrocyte CR1 (E-CR1) and glomerular CR1 (G-CR1) in different kinds of nephropathies using ELISA and immunofluorescence microscopy to understand their role in immune complex (IC) mediated renal diseases. E-CR1 was significantly reduced in all categories of lupus nephritis in comparison to normal subjects and non-IC renal diseases. However, other IC mediated diseases like IgA nephropathy and membranoproliferative glomerulonephritis had normal E CR1 levels. G-CR1 showed distinct differences between IC and non-IC mediated diseases. G-CR1 was virtually absent in lupus kidneys. In other IC mediated diseases, there was a correlation of G-CR1 expression to the IC and complement fragment deposition. G-CR1 serves as a useful diagnostic marker for IC mediated diseases while E-CR1 is useful as a prognostic marker to monitor the course of disease after the treatment has initiated. PMID- 11495297 TI - Hydroxychloroquine sulphate inhibits in vitro apoptosis of circulating lymphocytes in patients with systemic lupus erythematosus. AB - The serological hallmark of systemic lupus erythematosus (SLE) is the presence of antibodies against double-stranded DNA. However, several studies have suggested that it is not DNA itself, but nucleosomes that are the immunogenic particles involved both in the induction of anti-DNA antibodies, and in the pathophysiology of SLE. Meanwhile, It has been demonstrated that there is an accelerated in vitro apoptosis of lymphocytes from patients with SLE. Therefore, one can postulate that the process of apoptosis may provide a source of nuclear antigens to drive the autoantibody response seen in SLE. Our study has demonstrated that hydroxychloroquine exhibits an anti-apoptotic action and this anti-apoptotic effect is dependent on monocyte coexistence. We used both morphology assessment and fluorescent antibody cell sorter (FACS) analysis to measure the apoptotic percentage of lymphocytes from 25 SLE patients in medium alone (control) or with the addition of different concentrations of hydroxychloroquine. Our results have shown that there is a significant decrease in the percentage of apoptosis at the therapeutic concentration (10(-6) M) as compared with the control (p < 0.05). It has been reported that the anti-rheumatic properties of hydroxychloroquine result from its interference with antigen processing in macrophages and other antigen presenting cells. We propose that this results in decreased stimulation of autoreactive lymphocytes reactive with self-peptides, and consequently diminution of activation-induced cell death (apoptosis) of mature peripheral lymphocytes. PMID- 11495298 TI - Burkholderia pseudomallei-specific recombinant protein and its potential in the diagnosis of melioidosis. AB - Melioidosis is an important public health problem in Southeast Asia and Northern Australia. This disease is caused by the gram-negative bacilli, Burkholderia pseudomallei. Wide spectra of clinical manifestations are observed in melioidosis ranging from asymptomatic to septicemic infection. Although serodiagnostic methods of melioidosis have been improved significantly in recent years, a highly specific diagnostic test that can differentiate asymptomatic seropositive individuals and melioidosis patients remains to be the subject of current investigations. In this study, a B. pseudomallei-specific gene, pBps-1, expressing a novel 18.7 kDa recombinant protein was selected from genomic libraries of two B. pseudomallei virulent isolates by using pooled sera from septicemic melioidosis patients. Nucleotide sequence analysis demonstrated that this gene is unique and does not show substantial similarity with any known genes in the Genbank database. The Bps-1 recombinant protein was evaluated for its potential in serodiagnosis of melioidosis by Western blot analysis. A high degree of specificity was demonstrated using sera from healthy individuals in the endemic (98.5%) and non-endemic areas (100%), with moderate sensitivity (69.7%) in melioidosis patients. The study demonstrated that this approach can be used to obtain highly specific recombinant antigens such as that described in the present report. A combination of such antigens should provide materials for successful serodiagnosis of melioidosis in the endemic areas. PMID- 11495299 TI - Quantitation of human immunodeficiency virus type 1 DNA by competitive nested PCR assay. AB - A quantitative competitive nested PCR assay was developed for quantifying HIV-1 proviral DNA in clinical samples. A competitor DNA was constructed from a conserved region of the HIV-1 gag gene by deleting a sequence of 18 base pairs. We quantitated HIV-1 proviral DNA copy number in clinical samples. Peripheral blood mononuclear cells (PBMCs) from 35 HIV-infected patients with a CD4 count range of 4-728 cell/mm3 were analyzed by this method. The copy numbers of HIV-1 DNA detected ranged between 518 to 67,340 copies per 10(6) CD4+ T-cells. The copy numbers correlated inversely with the CD4 counts. PMID- 11495300 TI - Predominant human rotavirus genotype G1P[8] infection in infants and children in Bangkok, Thailand. AB - Human rotavirus is the major etiologic agent of infantile diarrhea on a worldwide scale. In this study, rotaviruses were detected by reverse-transcription PCR in 42 of 83 stool specimens from children below the age of 3 years with acute diarrhea in Bangkok, Thailand, between November 1998 and August 1999. G and P types of all samples were characterized by restriction endonuclease analysis (REA) and multiplex PCR typing assay, respectively. Strain G1P[8] (76.1%) was the predominant type, followed by G1P[6] (2.4%). Strain G1 combined with mixed P[8]/P[6] was identified in 2 specimens (4.8%) and 7 untypeable G strains (16.7%) were observed. This information on the circulating G and P combinations should be useful for understanding the epidemiology of human rotavirus in Bangkok, Thailand. PMID- 11495301 TI - Paradoxical vocal cord adduction mimicking as acute asthma in a pediatric patient. AB - We report an adolescent girl with paradoxical vocal cord adduction who presented with acute onset of hyperventilation, wheezing and stridor that did not respond to bronchodilator and anti-inflammation therapy. The paradoxical vocal cord motion was confirmed by flexible fiberoptic bronchoscopic examination. We found the stridor was induced by hyperventilation, and was caused by paradoxical vocal cord movement. The abnormal cord motion may be psychogenic and could be misdiagnosed as asthma. It is important to investigate the underlying background and social history and to avoid unnecessary use of beta-agonists, steroids, and even endotracheal intubation or tracheostomy. PMID- 11495302 TI - The effects of exogenous interleukin-12 on the induction of immune response to Porphyromonas gingivalis in vivo in mice. AB - The effects of treatment with exogenous interleukin-12 (IL-12) on the induction of immune response to Porphyromonas gingivalis, a black pigmented periodontopathic oral bacterium in mice, were determined in the present study. An increased footpad swelling representing a delayed type hypersensitivity (DTH) response to P. gingivalis in IL-12-treated mice could be observed, although increasing doses of IL-12 did not produce cumulative effects on this cellular Immune response. Multiple injections with IL-12 also resulted in elevated serum IFN-gamma levels. Treatment with this cytokine the day before, on and after immunization with heat-killed P. gingivalis augmented the levels of serum antigen specific IgG2a and IgG3 antibodies, but had obviously little or no effects on those of serum antigen-specific IgG1 and IgG2b antibodies. The results of this study suggest that treatment with exogenous IL-12 In P. gingivalis-immunized mice may enhance DTH response and Th1 cell-associated antibody production. PMID- 11495303 TI - The relevance of specific serum IgG, IgG4 and IgE in the determination of shrimp and crab allergies in Malaysian allergic rhinitis patients. AB - The significance of food specific serum IgG4 antibody in food allergy is unclear and this led us to investigate the relevance of specific IgG4, along with IgG and IgE antibodies to two common food allergens in Malaysia. Enzyme-linked immunosorbent assay (ELISA) was used to measure the serum antibodies in 143 allergic rhinitis patients' sera, of which 47 were from patients with clinical indication of shrimp allergy, 46 with clinical indication of crab allergy and 50 without indication to either allergy. Clinical indication of allergy was based on answers to a questionnaire or results of the skin prick test. We found that the elevation of specific IgE or IgG4 is associated with shrimp and crab allergies but elevation of specific IgG is not associated with either allergy. However, the clinical utility of elevated specific IgG and IgG4 levels is pending further investigation. PMID- 11495304 TI - Color vision in children. PMID- 11495305 TI - Preoperative local measures minimize bacteremia during eye surgery. PMID- 11495306 TI - Detection and assessment of color vision anomalies and deficiencies in children. PMID- 11495308 TI - Normal neonatal values of ophthalmic and central retinal artery blood flow velocities. AB - PURPOSE: To define standard values of blood flow velocities and indices in the ophthalmic and central retinal arteries in the neonatal period. METHODS: Forty two healthy full-term neonates comprised the study population. A color Doppler with mechanical sector probe was used for measuring blood flow velocity in the ophthalmic and central retinal arteries. Systolic, end diastolic, and mean enveloped velocities were measured, and the resistance index and pulsatility index were calculated. RESULTS: Ophthalmic artery Doppler velocities were similar on the first and third days of life, but increased significantly on the fifth and seventh days of life; resistance index significantly increased during the first week of life, whereas pulsatility index did not change significantly. Doppler velocities of the central retinal artery were similar on the first and third days; they show a delayed increase compared to the ophthalmic artery. Central retinal artery blood flow velocities increased significantly from the third to seventh postnatal day. Resistance index also increased between the first two days and on the fifth and seventh postnatal days, while pulsatility index did not change. CONCLUSION: These data constitute a starting point for studying the possible relationship between eye circulation and pathogenesis of retinopathy of prematurity. PMID- 11495307 TI - Diagnosis and management of chronic blepharokeratoconjunctivitis in children. AB - PURPOSE: To describe the history, symptoms, and clinical signs and discuss the treatment of blepharokeratoconjunctivitis. METHODS: Eight children (five girls and three boys), ranging in age from 3.5-13 years, were clinically diagnosed with blepharokeratoconjunctivitis. Microbiology studies were performed in four of the eight children. Treatment consisted of lid hygiene, oral erythromycin suspension, and preservative-free steroids. Duration of therapy was directed by clinical improvement. RESULTS: Average age at onset was 3.2 years (range: 0.5-8 years). Lid disease, conjunctival redness, and inferior superficial corneal vascularization were consistent features (100%). Other signs were punctate corneal epithelial staining, inferior subepithelial vascularization and infiltrate, conjunctival phlyctenules, corneal phlyctenules, and circumferential pannus. Microbiology testing demonstrated coagulase-negative staphylococcus and Propionibacterium acnes. Average follow-up was 8.3 months (range: 2-23 months). All patients had relief of symptoms within 2-3 weeks. Clinical signs took more time to regress but all had progressive improvement of the ocular surface by 2 months. Blepharokeratoconjunctivitis reactivated in all patients during follow up, and repeat therapy was administered. CONCLUSION: Blepharokeratoconjunctivitis in childhood is a chronic inflammatory process that can have different presentations. It can be successfully treated with oral erythromycin and topical steroids. PMID- 11495309 TI - Palpebral fissures changes in early childhood. AB - PURPOSE: To describe palpebral fissure changes during early childhood. METHODS: Digital imaging was used to analyze the monocular palpebral fissure images of 185 children and 35 adults. Six variables were quantified for each eye: the distance between the pupil center and the upper and lower eyelid margin, the palpebral fissure area, and the obliquity and length of the fissure. The lower eyelid crease pattern also was evaluated qualitatively. RESULTS: The upper eyelid was at its lowest position after birth, and the lower eyelid margin was close to the pupil center. Between ages 3 and 6 months, the position of the upper eyelid reached its maximum and then declined linearly. At the same time, the distance between the pupil center and the lower eyelid margin increased linearly until age 18 months when its position stabilized. Both the horizontal palpebral fissure length and the fissure area followed a logarithmic pattern of growth and were strongly correlated. The obliquity of the fissure was evident at an early age and did not change until adulthood. At birth, the most common pattern of the lower eyelid crease was a single crease. The number of individuals showing a double crease increased with age, and at approximately age 36 months, a double crease became the most common pattern. CONCLUSION: The palpebral fissure undergoes complex changes during infancy. An accurate knowledge of these changes is important for evaluating eyelid disorders in children. PMID- 11495310 TI - Iris neovascularization in children as a manifestation of underlying medulloepithelioma. AB - PURPOSE: To report the occurrence of unilateral iris neovascularization in children secondary to medulloepithelioma. METHODS: Presenting features and the clinical course of patients confirmed to have medulloepithelioma were reviewed. RESULTS: Seven patients with medulloepithelioma had iris neovascularization during the clinical course. Associated cataract and lens coloboma occurred in two and three cases, respectively. In six (86%) cases, no apparent cause for iris neovascularization could be detected at presentation. Two cases had tube shunts for management of neovascular glaucoma before medulloepithelioma was recognized. All cases eventually required enucleation. CONCLUSION: Presence of iris neovascularization is an early manifestation of medulloepithelioma. Children with iris neovascularization of unknown cause should be evaluated to exclude underlying medulloepithelioma. PMID- 11495311 TI - Neuro-ophthalmic features in pediatric tubercular meningoencephalitis. AB - PURPOSE: To examine the neuro-ophthalmic profile in childhood tubercular meningoencephalitis and assess its prognostic importance. METHODS: One hundred consecutive patients with tubercular meningoencephalitis underwent evaluation for optic nerve functions, supra- and infranuclear oculomotor involvement, and raised intracranial tension. Patients underwent follow-up for 9 months. RESULTS: In 100 patients with tubercular meningoencephalitis, 67 had neuro-ophthalmic features. Common findings were retrobulbar neuritis (32) and gaze palsy (20). Others, in decreasing prevalence, were third and sixth nerve palsy, conjugate deviation, primary optic atrophy, and complete ophthalmoplegia. Raised intracranial tension was associated with neuro-ophthalmic features in 53 of 67 patients (P<.05). Complete ophthalmoplegia was significantly associated with mortality on logistic regression analysis (relative risk, 6.35; P=.012). By 6 months, 56% of patients with retrobulbar neuritis progressed to optic atrophy. CONCLUSION: Neuro ophthalmic features, in association with raised intracranial tension, often are a part of pediatric tubercular meningoencephalitis presentation. The type of neuro ophthalmic features may have prognostic value. Optic nerve involvement is the most common finding in tubercular meningoencephalitis. PMID- 11495312 TI - Characteristics of Okihiro syndrome. AB - PURPOSE: To describe Okihiro syndrome, a syndrome consisting of Duane's syndrome, radial ray anomaly, and sensorineural deafness. METHODS: The incidence of anomalies in 41 individuals from 9 affected families was assessed by literature review and patient examination, and the frequency of the characteristics occurring together was determined. RESULTS: Of all the individuals with at least one of the above characteristics, only 12% had all three characteristics. However, 44% had both Duane's syndrome and radial ray anomaly, whereas sensorineural deafness alone occurred in only 17% of patients. CONCLUSION: Okihiro syndrome is consistent with autosomal dominance with variable penetrance, manifesting primarily with Duane's syndrome and radial ray anomaly, and less frequently, sensorineural deafness. PMID- 11495313 TI - Ocular involvement in childhood acute lymphoblastic leukemia. PMID- 11495314 TI - Microphthalmos with cyst: preservation of the eye by repeated aspiration. PMID- 11495315 TI - Retinoblastoma and the 13q deletion syndrome. PMID- 11495316 TI - Duane's syndrome in a patient with small intestine malrotation. PMID- 11495317 TI - The assessment of sport-related concussion: the evidence behind neuropsychological testing and management. AB - Because of the lack of valid evidence to support the current recommendations for the management of mild traumatic brain injury (concussion), many physicians, athletic trainers, coaches, and athletes have called into question the way concussions are treated in athletics. This review article discusses the current evidence for the management of concussion in high school, college, and professional sports. A complete review of the epidemiologic and neuropsychological studies to date is presented and critically reviewed, as are other assessment and management tools in concussion. The appropriate use of neuropsychological testing, grading scales, and return-to-play recommendations are discussed in depth based on the current evidence. Additionally, areas requiring further research are identified and future trends are briefly discussed. PMID- 11495318 TI - Does second impact syndrome exist? AB - Second impact syndrome (SIS) is a widely feared complication of traumatic brain injury. Although postulated to occur after repeated concussion, the evidence for such a premise is not compelling. This paper reviews the published evidence for and against the existence of this controversial entity. Rather than SIS being a complication of recurrent concussion, it is far more likely that the clinical condition represents "diffuse cerebral swelling," a well-recognized complication of traumatic brain injury. This condition is more common in children and adolescents, which reflects the known demographics of so-called "second impact syndrome." We propose that clinicians abandon the misleading term second impact syndrome and refer to the syndrome as diffuse cerebral swelling. PMID- 11495319 TI - Evidence-Based review of sport-related concussion: clinical science. AB - The clinical nature of sport-related concussion is discussed in this paper. Particularly highlighted are the difficulties with definition, injury severity grading, classification, and understanding of clinical symptoms. In addition, the well-recognized sequelae of concussion including the motor and convulsive manifestations are discussed in detail. Where possible, an evidence-based approach is adopted to assist the understanding of the literature in this complex area. PMID- 11495320 TI - Evidence-based review of sport-related concussion: basic science. AB - The evidence base for sport-related concussive brain injury is reviewed in this paper. In the past, pathophysiological understanding of this common condition has been extrapolated from studies of severe brain trauma. More recent scientific study demonstrates that this approach is unsatisfactory, and the clinical features of concussion represent a predominantly functional brain injury rather than manifest by structural or neuropathological damage. Such understanding of this condition remains incomplete at this stage. PMID- 11495321 TI - New frontiers in diagnostic imaging in concussive head injury. AB - Concussed athletes may have documented incapacitating postconcussive symptoms, neuropsychological deficits, and consequent important changes in their lives and sport, yet the majority of neuroimaging attempts reveal few findings to account for these signs and symptoms. In this paper, we explore new techniques in the neuroimaging of concussion including diffusion-weighted magnetic resonance imaging and functional brain imaging technology. PMID- 11495322 TI - Standardized mental status assessment of sports concussion. AB - Neurocognitive status is often considered the domain of neurologic functioning most sensitive to change following concussion, but the effects are often subtle and difficult to detect on routine clinical examination. Recent efforts have focused on the development of brief, standardized methods of mental status assessment for use by sports medicine clinicians to quantify the acute neurocognitive effects of concussion and objectively track postinjury recovery. Research has demonstrated the reliability, validity, and sensitivity of these measures in detecting concussion in athletes and providing empirical data for consideration in the context of other examination findings, neuropsychologic test data, and neuroimaging results. Standardized measures of mental status and other postconcussive symptoms are valuable tools to assist clinicians in the assessment and management of concussion, but should not be used as a replacement for medical evaluation or viewed as a stand-alone means for determining readiness to return to competition after injury. PMID- 11495323 TI - Postural stability assessment following concussion: one piece of the puzzle. AB - Clinicians regularly assess concussion according to the symptoms that an athlete manifests at the time of injury, as well as during subsequent evaluations. The subjectivity involved with symptom assessment, however, often leaves the clinician without a clear picture of the athlete's true mental status. Neuropsychologic testing has become very popular in the sports medicine community for assessing the cognitive domain of neurologic functioning, and postural stability testing is gaining credence for assessing the motor domain. The objective of this review was to determine the efficacy of postural stability testing as an adjunct to concussion assessment of athletes. Multiple studies, using both sophisticated force plate technology, as well as those using less sophisticated clinical balance tests, have identified postural stability deficits lasting several days following sport-related concussion. It appears that postural stability testing provides a useful tool for objectively assessing the motor domain of neurologic functioning, and should be considered a reliable and valid adjunct to the assessment of athletes suffering from concussion. Although symptom severity, neurocognitive function, and postural stability are often affected initially following concussion, they are not necessarily related or even affected to the same degree. PMID- 11495324 TI - New treatments for concussion: the next millennium beckons. AB - As increased understanding of the pathophysiology of mild traumatic brain injury and concussion develops, so the scientific rationale for interventional pharmacological therapy becomes paramount. A number of agents have been postulated or have been the subject of anecdotal noncontrolled trials. This paper reviews the published evidence in this regard. To date no effective pharmacological therapy exists that satisfies Class I evidence-based medicine criteria. PMID- 11495325 TI - The cumulative effect of repetitive concussion in sports. AB - The increased popularity of contact sports worldwide exposes a large number of participants to both acute and chronic traumatic brain injury. Chronic traumatic brain injury (CTBI) represents the cumulative, long-term neurological consequences of repetitive concussive and subconcussive blows to the brain. Although this condition has been described primarily in boxing, it may be anticipated in other contact sports such as soccer, football, ice hockey, and the martial arts. Since treatment options in CTBI are relatively limited, the prevention of CTBI is of paramount importance. Minimizing the frequency and severity of acute brain injury in sport will be instrumental in accomplishing this goal. The prevention of CTBI will need to be sport specific and will undoubtedly rely on limiting the exposure of high-risk athletes, utilizing of protective equipment, enforcing strict rule adherence, training and supervising athletes, and increasing medical surveillance. PMID- 11495326 TI - Legal issues affecting medical clearance to resume play after mild brain injury. AB - Physician care of athletes suffering mild brain trauma involves medical and legal considerations. Physicians must adhere to their primary responsibility to protect their athletes' health by following good medical practice and not allowing nonmedical factors to unduly influence their judgment. Courts recognize the team physician's role as gatekeeper and generally defer to his or her medical judgment in return-to-play decisions. PMID- 11495327 TI - Preseason conditioning to prevent soccer injuries in young women. PMID- 11495328 TI - Clinical research and the third world. PMID- 11495329 TI - Treating this heavy midlife of men. PMID- 11495330 TI - Constructing ethnicity: culture and ethnic conflict in the new world disorder. AB - The end of the Cold War saw a proliferation of deadly conflicts between ethnic and other so-called identity groups around the world. Building on the tension between the purely "cultural" and politically deployable "ethnic" dimensions of identity, this paper examines the emergence of these conflicts in terms of the disintegration of major power systems--postcolonial, postimperial, postrevolutionary, and postillusionary--that had hitherto operated to constrain conflict. The emergence of the refugee as a new cultural, as well as political, type is discussed as one precipitate of these identity-group conflicts. PMID- 11495331 TI - Resilience in ecosystemic context: evolution of the concept. AB - The evolution of the resilience literature across diverse social science disciplines over the past two decades is reviewed and a synthesis of recent findings is offered, suggesting that resilience is a multidetermined and ever changing product of interacting forces within a given ecosystemic context. Emerging constructions of the concept are examined, and a refined Working definition is proposed. Implications for research and practice are offered. PMID- 11495332 TI - Youth exposure to violence: prevalence, risks, and consequences. AB - Recent empirical work on the distribution, determinants, and consequences of children and adolescents' witnessing of community violence are reviewed. Major findings across studies indicate that males, ethnic minorities, and urban residents are at increased risk for witnessing violence, and that higher rates of PTSD, depression, distress, aggression, and externalizing behavior disturbances are reported among those who witness violence. Degree of family conflict, domestic violence, and family support were demonstrated to modify the impact of exposure to violence. Research and policy recommendations are offered. PMID- 11495333 TI - Understanding links among childhood trauma, dissociation, and women's mental health. AB - Interrelationships among pathological dissociation, child and adult trauma exposure, and adult mental health symptoms were examined in a sample of low income, mostly African-American women. Dissociation was significantly related to both trauma exposure and mental health symptoms but did not mediate this relationship. Implications for research and practice are discussed. PMID- 11495334 TI - Childhood physical abuse, personality, and adult relationship violence: a model of vulnerability to victimization. AB - An association between childhood physical abuse and current relationship violence was observed in a sample of 56 young adult women, suggesting that childhood physical abuse may be a unique risk factor for victimization in intimate adult relationships. Aspects of personality and interpersonal functioning that reflect extreme doubt about one's interpretation and understanding of interpersonal events, and an inability to make sensible causal connections between people's thoughts, feelings, and actions largely mediated this link. Implications for intervention and directions for future research are discussed. PMID- 11495335 TI - Variables in delayed disclosure of childhood sexual abuse. AB - In a study of 41 adult survivors of childhood sexual abuse, the level of childhood traumatization was found to have contributed to delayed disclosure of the abuse. Other delaying variables included: belief in the importance of obedience to grownups, mistrust of people, fear of social rejection, and fear of the criminal justice system. Variables such as media attention to similar cases and experiences of personal achievement were inversely related to the age at disclosure. Recommendations for policy are discussed. PMID- 11495336 TI - Relationship of young adult Chinese Americans with their parents: variation by migratory status and cultural orientation. AB - To examine whether Chinese and American cultural orientations mediate the association between migratory status and parent relationship, 122 American-born, 121 early-immigrant, and 110 late-immigrant Chinese young adults were measured on cultural orientation and parent relationship. The poorest relationships were found in the early-immigrant group. Cultural orientation mediated the difference in parent relationship between early and late immigrant groups, but not between early-immigrant and American-born groups. Implications of the findings for research and practice are discussed. PMID- 11495337 TI - Attribution and serious mental illness: understanding multiple perspectives and ethnocultural factors. AB - Attributions regarding the nature, cause, and best treatment of mental health problems were examined in a total of 563 adults with serious mental illness from four ethnocultural groups, 185 family members, and 43 mental health practitioners. Attributions agreed in many areas, such as stress as a cause, and medical intervention and counseling as best treatment. Differences were found between the practitioner and other groups in attributions regarding the senses in the nature of mental illness, and in drugs and alcohol as a cause. The role of ethnicity was examined and treatment implications discussed. PMID- 11495338 TI - Mental health and marital adaptation among mothers of children with cerebral palsy. AB - Employing a theoretical model that emphasizes risk-resistance factors for maternal adaptation, this study compared 80 mothers of children with cerebral palsy to 80 mothers of children without physical disabilities. Results revealed differences between the two groups, and indicated that, among mothers of children with cerebral palsy, self-esteem, self-mastery, and family network size were the main variables that contributed to mothers' psychological and marital adaptation. PMID- 11495339 TI - Posttraumatic stress symptoms following near-death experiences. AB - Persons who report "near-death experiences" (NDEs) acknowledge more intrusive symptoms of posttraumatic stress disorder (PTSD) than those who came close to death without NDEs, but not more avoidance symptoms, suggesting a nonspecific stress response. Although dissociation generally increases vulnerability to PTSD, the positive affect that distinguishes NDEs from other dissociative experiences may mitigate subsequent PTSD symptoms. PMID- 11495340 TI - Attachment style classification and posttraumatic stress disorder in former prisoners of war. AB - Adult attachment style and post-traumatic stress disorder (PTSD) symptomatology were investigated in 107 former prisoner of war veterans. Those with secure attachment styles scored significantly lower on measures of PTSD than did those with insecure styles, and attachment style was a stronger predictor of PTSD symptom intensity than was trauma severity. The suggested association between attachment style and PTSD's development and persistence are discussed in relation to research and clinical practice. PMID- 11495341 TI - Predictors of homicidal ideation and intent in schizophrenia: an empirical study. AB - Correlations of age, gender, and psychosocial variables with ratings of current homicidal ideation and intent were investigated among psychiatric facility clients with schizophrenia. Multiple regression analyses revealed, irrespective of age or gender, significantly positive correlations of three psychosocial variables--manic symptoms, psychotic symptoms, and impaired global functioning- with homicidal ideation and intent. Implications for research are discussed. PMID- 11495342 TI - An HIV/AIDS risk-reduction program for mentally ill hospital patients: assessing readiness for change. AB - This study evaluated the effectiveness of an HIV/AIDS cognitive-behavioral skills training program in promoting readiness for behavior change among a sample of psychiatric inpatients. Significant differences were found in the domain of precontemplation for women patients, in domain of contemplation for patients diagnosed with a psychotic disorder, and in the domain of action for men and for patients with a mood disorder. In all areas, difference scores reflected an improvement between pre- and posttest levels. PMID- 11495343 TI - H2O2-induced cell death in human glioma cells: role of lipid peroxidation and PARP activation. AB - Reactive oxygen species (ROS) have been implicated in the pathogenesis of a number of neurodegenerative disorders. However, the underlying mechanism of ROS induced cell injury remains to be defined. This study was undertaken to examine the role of lipid peroxidation and poly (ADP-ribose) polymerase (PARP) activation in H2O2-induced cell death in A172 cells, a human glioma cell line. H2O2 induced a dose- and time-dependent cell death. The cell death was prevented by thiols (dithiothreitol and glutathione), iron chelators (deferoxamine and phenanthroline), H2O2 scavengers (catalase and pyruvate), and a hydroxyl radical scavenger (dimethylthiourea). Antioxidants N,N'-diphenyl-p-phenylenediamine (DPPD) and Trolox had no effect on the H2O2-induced cell death. Lipid peroxidation did not increase in human glioma cells exposed to H2O2. The PARP inhibitor 3-aminobenzamide prevented the cell death induced by H2O2. The PARP activity was increased by H2O2 and the H2O2 effect was prevented by 3 aminobenzamide, dithiothreitol, and phenanthroline. The ATP depletion induced by H2O2 was prevented by catalase, dithiothreitol, phenanthroline, and 3 aminobenzamide, but not by DPPD. These results indicate that the H2O2-induced cell death is mediated by PARP activation but not by lipid peroxidation in human glioma cells. PMID- 11495344 TI - D4 dopamine and metabotropic glutamate receptors in cerebral cortex and striatum in rat brain. AB - The characterization of the functional interactions between the metabotropic glutamate receptors (mGluR) and the dopaminergic (DR) receptors in the corticostriatal projections may provide a possible interpretation of synaptic events in the basal ganglia. It has been suggested that presynaptic D2-type receptor located on glutamatergic corticostriatal neurons regulates the release of glutamate. In a first approach we have studied the cellular distribution of the D4R and the mGluRs in cerebral cortex and striatum employing immunocytochemistry. D4R positive neurons were particularly numerous in medial prefrontal cortex mainly occupying layers II and III. An even distribution was found on small round-shaped neurons in the striatum. Group I mGluR1alpha-like immunoreactivity (mGluR1alpha-LI) was found in medial and deep layers of the cerebral cortex while group III mGluR4a labeled more superficial layers; group II mGluR2/3 signal was intense on fine fibers with a punctate appearance. In the striatum, mGluR1alpha and mGluR2/3 stained mainly fibers while mGluR4a labeled round shaped cell bodies. After lateral ventricular injection of colchicine, an axonal transport and firing activity blocker, D4R labeling significantly increased in cerebral cortex and decreased in the striatum. mGluR1alpha and mGluR4a signal decreased in cerebral cortex and only mGluR4a signal decreased in the striatum. These results support previous reports indicating a presynaptic localization of D4R in the striatum. In contrast, striatal mGluR1alpha appears to be a postsynaptic receptor probably synthesized in situ. Our results do not support the hypothesis of a colocalization of D4 receptor and one or more of the metabotropic glutamatergic receptors studied here. PMID- 11495345 TI - The cerebrocortical areas in normal brain aging and in Alzheimer's disease: noticeable differences in the lipid peroxidation level and in antioxidant defense. AB - The markers of oxidative stress were measured in four cerebrocortical regions of Alzheimer's disease (AD) and age-matched control brains. In controls the levels of diene conjugates (DC) and lipid peroxides (LOOH) were significantly higher in the sensory postcentral and occipital primary cortex than in the temporal inferior or frontal inferior cortex. The antioxidant capacity (AOC) was highest in the temporal, and GSH in the frontal inferior cortex. The highest activity of superoxide dismutase (SOD) and catalase (CAT) was found in the occipital primary cortex. Compared with controls, significantly higher level of DC and LOOH and attenuated AOC were evident in AD temporal inferior cortex. In AD frontal inferior cortex moderate increase in LOOH was associated with positive correlation between SOD activity and counts of senile plaques. Our data suggest that in AD cerebral cortex, the oxidative stress is expressed in the reducing sequence: temporal inferior cortex > frontal inferior cortex > sensory postcentral cortex approximately = occipital primary cortex, corresponding to the histopathological spreading of AD from the associative to primary cortical areas. PMID- 11495347 TI - Effect of hypothermia on brain cell membrane function and energy metabolism in experimental Escherichia coli meningitis in the newborn piglet. AB - We evaluated the anti-inflammatory and neuroprotective effects of hypothermia during the early phase of experimental Escherichia coli meningitis in the newborn piglet. Hypothermia significantly attenuated the meningitis-induced acute inflammatory responses such as increased intracranial pressure, decreased glucose level, increased lactate concentration, increased tumor necrosis factor-alpha level and leukocytosis in the cerebrospinal fluid. Decreased cerebral cortical cell membrane Na+,K+-ATPase activity and increased lipid peroxidation products, indicative of meningitis-induced brain damage, were significantly improved with hypothermia. Hypothermia also significantly improved the meningitis-induced reduction in brain ATP and phosphocreatine levels. In summary, hypothermia significantly attenuated the acute inflammatory responses and the ensuing brain injury in experimental neonatal bacterial meningitis. PMID- 11495346 TI - Subchronic treatment with methamphetamine and phencyclidine differentially alters the adenosine A1 and A2A receptors in the prefrontal cortex, hippocampus, and striatum of the rat. AB - Subchronic treatment with MAP (4.6 mg/kg, i.p., once daily for 11 days) significantly decreased the Kd, but not Bmax, values of [3H]1,3-dipropyl-8 cyclopentylxanthine ([3H]DPCPX) binding to adenosine A1 receptors in the prefrontal cortex and hippocampus, but not striatum, of rat brain. However, subchronic treatment with PCP (10 mg/kg, i.p., once daily for 11 days) did not alter the Kd and Bmax values of [3H]DPCPX binding to adenosine A1 receptors in these three regions. Subchronic treatment with MAP or PCP did not alter the Bmax and Kd values of [3H]2-p-(2-carboxyehyl)phenethylamino-5'-N ethylcarboxyamidoadenosine ([3H]CGS21680) binding to adenosine A2A receptors in the striatum. Furthermore, subchronic treatment with MAP or PCP significantly decreased the specific binding of [3H]CGS21680 to adenosine A2A receptors in the hippocampus, but not in the prefrontal cortex. Thus, these results suggest that MAP and PCP may produce differential effects on the adenosine A2A receptors, but not adenosine A1 receptors in rat brain. PMID- 11495348 TI - GM2 ganglioside regulates the function of ciliary neurotrophic factor receptor in murine immortalized motor neuron-like cells (NSC-34). AB - We previously reported that ciliary neurotrophic factor (CNTF) increased the serum-free cell survival of immortalized motor neuron-like cells (NSC-34), and addition of the exogenous ganglioside GalNAc beta4(Neu5Ac alpha3)Gal beta4GlcCer (GM2) facilitated cell survival together with CNTF. Moreover beta 1,4 N acetylgalactosaminyltransferase (GM2 synthase) activity increased in NSC-34 cells cultured with CNTF. We now have examined whether CNTF-induced cell survival is associated with the collaboration between GM2 and the CNTF receptor (CNTF-R). Despite the presence of CNTF (50 ng/ml), anti-CNTF-R antibody caused cell death and prevented the up-regulation of GM2 synthase expression. The addition of GM2 (1 to 20 microM) abrogated the anti-CNTF-R antibody effect which shortened cell survival and blocked GM2 synthase activation. Use of [125I]CNTF showed the specificity of CNTF binding in NSC-34 cells in situ. GM2 produced a 5-fold increase in the CNTF binding affinity per cell but did not change the binding site number. The study by metabolic labeling with [1-(14)C]N-acetyl-D galactosamine ([14C]GalNAc) showed that biosynthesized GM2 was involved in the immunoprecipitation of CNTF-R. These findings indicate that up-regulated GM2 synthesis induces functional conversion of CNTF-R to the activated state, in which it has affinity for CNTF. We conclude that GM2 is a bio-regulating molecule of CNTF-R in motor neurons. PMID- 11495349 TI - Interactions between allosteric modulators and 4-DAMP and other antagonists at muscarinic receptors: potential significance of the distance between the N and carboxyl C atoms in the molecules of antagonists. AB - Allosteric enhancement of the affinity of muscarinic receptors for their ligands offers a new way to influence cholinergic neurotransmission. The structure of the allosteric binding domain(s) and the features of agonists, antagonists and modulators which determine the occurrence of either positive or negative cooperativity require clarification. We tested interactions between allosteric modulators alcuronium, strychnine and brucine and eight antagonists at muscarinic receptors expressed in CHO cells. In experiments with unlabeled antagonists, all three modulators enhanced the affinity for 4-diphenylacetoxy-N dimethylpiperidinium (4-DAMP) at the M2 receptors, and strychnine did so also at the M4 receptors. Positive interactions were also observed between alcuronium and L-hyoscyamine (M2) and scopolamine (M2), between strychnine and butylscopolamine (M4), L-hyoscyamine (M2 and M4) and scopolamine (M4), and between brucine and scopolamine (M2). Positive effects of alcuronium, strychnine and brucine on the affinity of the M2 receptors for 4-DAMP have been confirmed by direct measurements of the binding of [3H]-4-DAMP. A comparison of molecular models of several antagonists which are esters revealed that antagonists in which the distance between the N and the carboxyl C atoms corresponds to five chemical bonds are more likely to display positive cooperativity with alcuronium at the M2 receptors than the antagonists in which the N-carboxyl C distance corresponds to four chemical bonds. PMID- 11495350 TI - A direct chemical interaction between dynorphin and excitatory amino acids. AB - The endogenous opioid peptide dynorphin A elicits non-opioid receptor-mediated neurotoxic effects. These effects are blocked by pretreatment with N-methyl-D aspartate (NMDA) receptor antagonists. Herein, the mechanism for the non-opioid effects of dynorphin and related peptides was studied by matrix-assisted laser desorption ionization (MALDI) mass-spectrometry. We observed that both glutamate or aspartate bind non-covalently to dynorphin A and dynorphin 2-17. However, when dynorphin A or dynorphin 2-17 were added to an equimolar mixture of Glutamate and Aspartate, they both complexed preferentially with glutamate. These data may explain the non-opioid physiological effects of dynorphin A and related peptides and indicate that the direct chemical interaction between neurotransmitters should be monitored when studying interactions between different neurochemical systems. PMID- 11495351 TI - Relationship between locomotor activity and monoamines following single and double transient forebrain ischemia in gerbils. AB - The relationship between locomotor activity and monoamine levels in gerbils after single and/or double forebrain ischemic insult was studied. Locomotor hyperactivity was observed after the first ischemic episode, but the gerbils failed to show hyperactivity after the second ischemic episode induced one week later. The monoamine levels were determined in order to clarify the biochemical basis of post-ischemic locomotor hyperactivity. Norepinephrine increased in response to first ischemic episode but remained at normal levels after the second episode of ischemia. Metabolites of dopamine and serotonin increased after both the first and second ischemic insults, which indicates that these monoamines do not play significant roles in post-ischemic locomotor activity. Therefore, increases in norepinephrine after first ischemic insult may play a role in increasing locomotor activity during the period following such an episode. PMID- 11495352 TI - Effects of fetal bovine serum on ferrous ion-induced oxidative stress in pheochromocytoma (PC12) cells. AB - Ferrous ion (Fe2+) has been considered to be a cause of neuronal oxidative injury. Since body fluids contain protein and serum is an essential component of tissue culture medium, we have examined the role of serum protein on Fe2+ mediated oxidative stress using PC12 cells and rat cerebral cortices. Fe2+ or the combination of ascorbate and Fe2+ increased concentrations of thiobarbituric acid reactive substances (TBARS) in PC12 cells and cerebrocortical homogenates in medium (RPMI 1640), but did not increase TBARS when the medium was supplemented with 10% fetal bovine serum. Treatment with ascorbate/Fe2+ in serum-free medium reduced endogenous glutathione (GSH) concentration in PC12 cells. However, the medium supplemented with serum did not reduce GSH concentrations. PC12 cell death induced by ascorbate/Fe2+ was alleviated by increasing serum or bovine albumin concentrations in the medium. These observations indicated that oxidative injury caused by the transition metal ion could be lessened by adding fetal bovine serum to culture medium. PMID- 11495353 TI - Polyamine oxidase activity in sera of depressed and schizophrenic patients after ECT treatment. AB - High level of polyamine oxidase activity is detected in sera of depressed as well as in schizophrenic patients. ECT treatment of depressed and schizophrenic patients reduced significantly the level of polyamine oxidase activity in their sera. After ECT treatment, clinically improved depressed and schizophrenic subjects were found to have sera polyamine oxidase activity not significantly differ from that of normal subjects. Possible biochemical mechanisms, which link polyamine oxidase activity, schizophrenia, depression and ECT effect are discussed here. PMID- 11495354 TI - Comparative analysis of superoxide dismutase activity between acute pharmacological models and a transgenic mouse model of Huntington's disease. AB - We examined the activity of striatal superoxide dismutase (SOD) in two acute pharmacological models of Huntington's disease (HD), and compared it with SOD activity in the striata of mice transgenic for the HD mutation. Total SOD, and Cu/ZnSOD activities increased in young transgenic mice, but decreased in older (35 week) mice. We consider that the increased enzyme activity represents a compensatory mechanism to protect cells from free radical-induced damage, but the system becomes insufficient in older animals. Major decreases in SOD activity were also observed both after quinolinic acid and 3-nitropropionic acid intrastriatal injections. The present results indicate that in both types of HD models striatal oxidative damage occurs, and that it is associated with alterations in the cellular antioxidant system. PMID- 11495355 TI - Multiple forms of phosphatase from human brain: isolation and partial characterization of affi-gel blue nonbinding phosphatase activities. AB - Phosphatases extracted from a human brain were resolved into two main groups, namely affi-gel blue-binding phosphatases and affi-gel blue-nonbinding phosphatases. Affi-gel blue binding phosphatases were further separated into four different phosphatase activities, designated P1-P4, and described previously. In the present study we describe the affi-gel blue-nonbinding phosphatases which were separated into seven different phosphatase activities, designated P5-P11 by poly-(L-lysine)-agarose and aminohexyl Sepharose 4B chromatographies. These seven phosphatase activities were active toward nonprotein phosphoester. P7-P11 and to some extent P5 could also dephosphorylate a phosphoprotein. They displayed different enzyme kinetics. On the basis of activity peak, the apparent molecular mass as estimated by Sephadex G-200 column chromatography for P5 was 49 kDa; P6, 32 kDa; P7, 150 kDa; P8, 250 kDa; P9, 165 kDa; P10, 90 kDa and P11, 165 kDa. Immunoblot analysis indicated that P8-P11 may belong to PP2B family, whereas P7 may associate with PP2A. The phosphatases P7-P11 were found to be effective in the dephosphorylation of Alzheimer's disease abnormally hyperphosphorylated tau. The resulting dephosphorylated tau regained its activity in promoting the microtubule assembly, suggesting that P7-P11 might regulate the phosphorylation of tau protein in the brain. PMID- 11495356 TI - Role of brain regional GABA: aldrin-induced stimulation of locomotor activity in rat. AB - Aldrin, a chlorinated hydrocarbon group of pesticide, is a well known central nervous system (CNS) stimulant. The CNS stimulating effect of aldrin is manifested in the form of an increase in locomotor activity (LA) of animals. Maximum increase in LA was observed at 2 h following aldrin (2-10 mg/kg, p.o.) treatment and this aldrin-induced increase in LA attained a peak at a dose of 10 mg/kg, p.o. Administration of aldrin (2 or 5 mg/kg/day, p.o.) enhanced LA of rats and reached a maxima after 12 consecutive days of treatment following which aldrin-induced LA was gradually reduced and restored to control value after 20 consecutive days of aldrin treatment. A single administration of aldrin (2-10 mg/kg, p.o.) reduced the GABA system in cerebellum, hypothalamus and pons medulla. The treatment with aldrin (2 mg/kg/day, p.o.) for 12 consecutive days produced more inhibition in those brain regional GABA system than that observed with a single dose of aldrin. These results, thus, suggest that aldrin-induced inhibition of central GABA may be a cause of stimulation of LA with aldrin either at a single dose or for 12 consecutive days. PMID- 11495357 TI - Amyloid beta peptide processing, insulin degrading enzyme, and butyrylcholinesterase. AB - Amyloid beta peptide implicated in Alzheimers disease is cleaved by insulin degrading enzyme (IDE). Abnormal cholinesterases similar to butyrylcholinesterase (BChE) are found in Alzheimer brain. The similarities between IDE and BChE (which is known to have an arylacylamidase and a metallocarboxypeptidase-like activity) such as their zinc metalloenzyme nature, their localization in glia and their ability to bind amyloid peptide in Alzheimers disease raise interesting questions. PMID- 11495358 TI - The development of interpersonal relationships in individuals with schizophrenia. AB - Three research assistants developed 2-week-long relationships with 15 patients with schizophrenia hospitalized at an inpatient psychiatric facility. Research assistants were assigned to no more than six patients during any one 2-week period. At the beginning and end of each 2-week relationship period, interactions between research assistants and patients were videotaped, and research assistants' negative and positive responses to patients were measured. Interaction partners' negative responses to patients increased over time. There were stable individual differences among patients in the degree to which they were liked by the research assistants and in how frequently research assistants made negative comments about patients. There were individual differences among research assistants in the degree to which they responded positively to patients. Patient strangeness and lack of pleasant conversational content were associated with the negative responses of research assistants. PMID- 11495359 TI - Barriers to relationship formation in schizophrenia: implications for treatment, social recovery, and translational research. PMID- 11495360 TI - Group psychotherapy with older adults. AB - This article describes a psychodynamically oriented psychotherapy group for older adults conducted in an agency setting, highlighting the problems specific to such groups. The literature on such groups for older adults is reviewed. The issues discussed are the psychology of late life, diagnostic considerations, medical illness and dementia as complicating factors, the issue of caretaking, and the centrality of shame. A systems-oriented approach to understanding the group's success is suggested. PMID- 11495361 TI - Ethical dilemmas, good intentions, and the road to hell: a clinical-ethical perspective on Yalom's depiction of Trotter's therapy. AB - Fiction in literature can assist in the analysis of therapeutic and ethical dilemmas in psychotherapy. Lying on the Couch (Harper Collins, 1997), Yalom's account of Dr. Seymour Trotter and Mrs. Belle Felini's therapeutic encounter, describes a complex series of issues and dilemmas. In the present article a reanalysis of the case from the perspective of a clinical and ethical consultation is presented. The therapeutic crisis that preceded the sexual relationship and the initial presentation of the client are considered in detail. Following the discussion of each of these periods, alternative modes of understanding and intervening in the treatment are presented. The in-depth exploration of a fictional case such as this one allows therapists to critically evaluate the decisions that underlie problems in the therapeutic encounter. The article concludes with specific recommendations for clinical and ethical practice. PMID- 11495362 TI - A critical look at the assumptions of cognitive therapy. AB - The cognitive perspective has been a dominant force in psychology and psychotherapy since the 1970s. Cognitive therapists assume that changing cognition is the key to bringing about changes in emotional reactions and symptomatic behaviors. This article examines evidence that is not consistent with the assumptions of cognitive therapy. First, neurobiological evidence is not consistent with the assumption that cognitions are necessary for emotions. Limbic structures allow for the possibility that emotions can be experienced without cortical processing. Second, research indicates that experiences can be stored as isolated affective fragments that distort cognitive functions. The assumptions of contemporary psychoanalytic models of therapy are consistent with this evidence. PMID- 11495363 TI - Changes in appearance and psychosis. PMID- 11495364 TI - Culture and eating disorders: a historical and cross-cultural review. AB - Cultural beliefs and attitudes have been identified as significant contributing factors in the development of eating disorders. Rates of these disorders appear to vary among different racial/ethnic and national groups, and they also change across time as cultures evolve. Eating disorders are, in fact, more prevalent within various cultural groups than previously recognized, both within American ethnic minorities and those in other countries. This review examines evidence for the role of culture as an etiological factor for the development of eating disorders. Historical and cross-cultural experiences suggest that cultural change itself may be associated with increased vulnerability to eating disorders, especially when values about physical aesthetics are involved. Such change may occur across time within a given society, or on an individual level, as when an immigrant moves into a new culture. Further research into the cultural factors that promote the development of eating disorders is much needed. Understanding how cultural forces contribute to the development of disorders is needed so that preventive interventions can be created. PMID- 11495365 TI - Complicating causes. PMID- 11495366 TI - Spiritual assessment: a review of major qualitative methods and a new framework for assessing spirituality. AB - This article introduces a new qualitative spiritual assessment instrument. It reviews existing qualitative assessment tools and presents a new multidimensional spiritual assessment framework. The instrument consists of two components: a spiritual history in which consumers relate their spiritual life story in a manner analogous to a family history and an interpretive framework to assist practitioners in eliciting and synthesizing the full potentiality of strengths extant in clients' spiritual lives. Common spiritual strengths the framework is designed to evoke are discussed, and a number of interventions based on prevalent spiritual strengths are suggested. PMID- 11495367 TI - Socialization in changing cultural contexts: A search for images of the "adaptive adult.". AB - In recent years increasing attention has been paid to the significant roles of culture and socialization in child development. The conceptual framework presented in this article is derived from the fields of cross-cultural child development, immigration, and ethnic minority studies. The article focuses on the relationship between cultural contexts and images of an "adaptive adult," which is proposed as a guiding metaphor in the organization of child-rearing ideologies and practices, which in turn are influenced by the values, experience, culture, and behavior of socializing agents within a given culture. The article explains how this concept is rooted in past and future time orientations and in the duality of public and private cultures. The article also explores changes in images of the adaptive adult in interaction with ecological components. Examples from the authors' research in Israel, as well as the research of others, are cited. The article concludes with a general discussion of the potential contribution that this framework can make to social workers dealing with immigrant, migrant, and ethnic minority children and families. PMID- 11495368 TI - Representation of ethnic identity in North American social work literature: a dossier of the Chinese people. AB - Ethnic and cultural identities of people who are not white in North America are conceived as natural and fixed categories. Such conceptualizations are associated with a tendency to take ethnicity as a client characteristic instead of understanding ethnic and cultural differences as constituted by the engagement between social worker and client. Using Foucault's dossier approach, the author uses the Chinese people as a case example to illustrate the politics of identification and identity assignment in professional social work literature in North America. The literature was selected from the Social Work Abstracts database from 1977 to 1997. The article reveals how Chinese people are "essentialized," "otherized," and negatively positioned as an ethnic construct. Four major arguments are presented together with their implications for cross cultural social work practice. PMID- 11495369 TI - Cultural competence for transracial adoptive parents. AB - This article provides a clear conceptual definition of cultural competence for transracial-cultural adoptive (TRA) parents based on an extensive review of the literature and feedback from both experts and parents. Following the differentiation of cultural competence as defined in the social work literature and cultural competence as applied to TRA parents, a three-part definition of cultural competence for TRA parents is presented. The article expands on each of three constructs: racial awareness, multicultural planning, and survival skills. In addition, it describes the process of beginning to operationalize the constructs. Finally, implications for social work practice, education, and research are suggested. PMID- 11495370 TI - The effects of poverty on childrens socioemotional development: an ecological systems analysis. AB - Bronfenbrenner's process-person-context-time model is used to examine theories that explain the adverse effects of economic deprivation on children's socioemotional development. In his model, each of five structures of the ecological environment-microsystems, mesosystems, exosystems, macrosystems, and chronosystems-is subsumed within the next higher level. Theories of the effects of poverty on proximal processes in the microsystem of the family have the most research support, but processes in other microsystems such as the peer group and school and in other levels of the ecological environment may also explain the relation between economic deprivation and children's socioemotional functioning. Social work practice and policy implications are drawn from the analysis. PMID- 11495371 TI - In-home continuing care services for substance-affected families: the bridges program. AB - Addressing substance abuse in families is an important concern for the social work field. This article presents a preliminary view of a continuing care substance abuse recovery services program designed to assist the substance affected family. The intervention approach is a blended model of substance abuse recovery work and family preservation. Services are directed at helping substance abusing parents with "recovering" their role with their families, developing support for their recovery work, and helping them gain the education and skills they need for effective parenting, supportive family involvement, and avoidance of drugs and alcohol. The program focuses on helping substance abusers and their families achieve relapse prevention by addressing functioning in four domains: individual actions and cognitions, individual recovery actions, family actions and cognitions, and family recovery actions. The article presents two case examples to highlight the efficacy of the intervention model and the general positive effect continuing care services are having on substance-affected families. PMID- 11495372 TI - Strengths-based case management: individuals' perspectives on strengths and the case manager relationship. AB - Strengths-based practice in social work has a strong theoretical foundation as an effective helping strategy that builds on a person's successes. Although there is growing empirical evidence informing outcomes associated with strengths-based approaches, missing from the literature is an understanding of how individuals who receive these services view their experiences. Qualitative data collection methods were used to gather individuals' experiences of participating in strengths-based case management implemented in a substance abuse aftercare program. The research questions that guided the study were "What are individuals' perceptions of strengths-based case management?" and "How do those perceptions compare and contrast to the key principles of strengths-based case management?" The emerging themes centered on individuals' responses to a focus on strengths (acceptance of strengths; holding on to strengths and deficits simultaneously; and initial mistrust of the approach) and to the relationship with the case manager (acceptance of the relationship; guilt when success is not achieved; and not needing the relationship). Implications for social work practice are discussed. PMID- 11495373 TI - Introduction: mass screening, health technology assessment, and health policy in some European countries. AB - OBJECTIVE: The series of papers in this issue was developed to examine the use of health technology assessment in policies toward prevention-specifically toward mass screening-in European countries. The papers actually examined three screening strategies: mammography screening for breast cancer, prostate-specific antigen screening for prostate cancer, and routine ultrasound in normal pregnancy. METHODS: Papers were sought from the member states of the European Union, plus Switzerland. Ultimately, nine acceptable papers were received, and were reviewed, revised, and edited. RESULTS: Screening is an accepted strategy in many countries for reducing the burden of disease through early detection and intervention. In part, this is because of successful screening programs that have been evaluated and implemented in many countries. At the same time, unevaluated and even useless and harmful screening programs-unjustified medically or economically-are widespread. Health technology assessment could help assure that only effective and cost-effective screening programs are implemented. CONCLUSION: The main conclusion is that screening is an important preventive strategy. Any screening program, however, should be carefully assessed before implementation. PMID- 11495374 TI - The accuracy and effectiveness of routine population screening with mammography, prostate-specific antigen, and prenatal ultrasound: a review of published scientific evidence. AB - OBJECTIVE: To review published data regarding the accuracy and effectiveness of three screening tests: mammography, prostate-specific antigen (PSA), and prenatal ultrasound. METHODS: Published evidence regarding the accuracy and effectiveness of the three tests was collected by computerized literature search and supplemented by manual review of relevant bibliographies. RESULTS: Screening mammograms lower breast cancer mortality by about 20%. Most data come from women aged 50-64 years; women aged 40-49 years may also benefit, but the absolute risk reduction is lower. Up to 1,500 to 2,500 women must undergo screening to prevent one death from breast cancer. Mammograms miss approximately 12% to 37% of cancers, generate false-positive results, and cause anxiety while abnormal results are evaluated. PSA screening can detect 80% to 85% of prostate cancers but has a high false-positive rate. There is little direct evidence that early detection reduces morbidity or mortality. Indirect evidence includes a trend toward earlier stage tumors and steadily declining mortality rates in geographic areas where PSA screening has become common. Potential harms include the morbidity associated with evaluating abnormal results, and complications from treatment (e.g., impotence, incontinence). The overall balance of benefits and harms remains uncertain in the absence of better evidence. Prenatal ultrasound may reduce perinatal mortality, primarily through elective abortions for congenital anomalies, but does not appear to lower live birth rates. Although ultrasound has no proven effect on neonatal morbidity, it provides more accurate estimates of gestational age that prevent unnecessary inductions for post-term pregnancy. Screening detects multiple gestations, congenital anomalies, and intrauterine growth retardation, but direct health benefits from having this knowledge are unproved. Ultrasound has both positive and negative psychological effects on parents. The scans do not appear to harm childhood development. CONCLUSIONS: Even for the most established screening tests, the appropriateness of routine testing depends on subjective value judgments about the quality of supporting evidence and about the trade-offs between benefits and harms. Individuals, clinicians, policy makers, and governments must weigh the evidence in light of these values and the constraints imposed by available resources. PMID- 11495375 TI - Screening in Austria: the cases of mammography, PSA testing, and routine use of ultrasound in pregnancy. AB - OBJECTIVE: To study cases of screening in Austria to learn about national strategies to handle the health policy challenge of early detection of widespread diseases and about the outcome of those strategies. The article describes three healthcare interventions (mammography, PSA testing, and routine use of ultrasound in pregnancy) and the instruments of Austrian health policy that are used-with or without explicit intention-to enforce or to control the widespread use of (early) diagnoses. METHODS: Data and information collection on healthcare services, their accessibility, rate of use, expert consensus, and official regulations. For all three case studies, expert interviews were carried out with main actors. RESULTS: Risk-group screening is not a priority in Austrian federal health policy. Although health promotion and prevention is a national task, examinations for early detection of specific diseases (i.e., carcinoma) are left to the health insurance funds, which delegate the decision to offer early diagnoses to their contracted physicians. In this opportunistic screening, general practitioners or specialists are encouraged by their health insurance funds or motivated by professional guidelines to offer certain examinations. CONCLUSIONS: Screening is a coordinated effort to acquire a grasp of a common disease at an early stage in a specified population. To achieve this objective, a culture of coordination and centralization has to be implemented. The collection of data is an essential element in coordination of decentralized medical interventions as much as quality control is an essential task in looking at and comparing the outcome of interventions. In the three case studies, neither of these two essential criteria were met. Evaluations and scientific evidence on the effectiveness of interventions were not used. PMID- 11495376 TI - Impact of health technology assessment on preventive screening in Belgium: case studies of mammography in breast cancer, PSA screening in prostate cancer, and ultrasound in normal pregnancy. AB - OBJECTIVE: To describe how scientific evidence has influenced healthcare policy making in Belgium in the field of sickness prevention for mammography, PSA testing in prostate cancer screening, and use of ultrasound in pregnancy. METHODS: Review of published and gray literature and interviews with stakeholders and experts. RESULTS: At the end of 1999, a systematic national/regional screening program had not yet been implemented for any of the three screening strategies. A systematic breast cancer screening program is being prepared for implementation only in Flanders. This limited impact can be attributed to the fragmentation in healthcare policy, the different options among the different regions, fragmentation in healthcare practice, the strong impact of healthcare stakeholders (provider groups and sickness funds) on decision making, and limited attention to scientific evidence in health policy and technology assessment. CONCLUSIONS: Health technology assessment has had very little impact on policy and practice in use of mammography, PSA testing, and ultrasound in pregnancy in Belgium. PMID- 11495377 TI - Evaluation of preventive technologies in Germany: case studies of mammography, prostate cancer screening, and fetal ultrasound. AB - OBJECTIVE: In this article, three preventive strategies-mammography screening for breast cancer, PSA screening for prostate cancer, and routine ultrasound in normal pregnancy-are discussed in the context of German health care. METHODS: Epidemiologic data and German studies evaluating different aspects of these preventive measures were identified and analyzed. RESULTS: Only a few studies could be identified that investigate these preventive measures. Despite sufficient evidence, in part derived from a German study, there is not yet a mammography screening program. In contrast, ultrasound in pregnancy is offered routinely, although there are controversies regarding the benefit of this practice. PSA screening is not offered as part of the screening program for prostate cancer. However, PSA tests as well as mammographies are done in large numbers in German ambulatory care-a practice that could be considered wild or opportunistic screening. CONCLUSIONS: These case studies show that preventive programs and practices in Germany are not sufficiently based on sound evidence. The paucity of evaluation activities related to prevention in Germany is probably due to the low threshold to introduce new preventive programs into the German healthcare system in the past. PMID- 11495378 TI - Health technology assessment in the area of prevention: selected screening cases in Greece. AB - OBJECTIVE: To explore the impact of health technology assessment (HTA) on health policy and practice in Greece through selected screening case studies in the prevention area. The three cases studied were mammography screening, PSA screening, and routine ultrasonography in normal pregnancy. METHODS: Official policy recommendations or reports, a literature review of Greek published research as well as gray literature from various sources, and interviews with specialists and medical associations were performed, and their impact on health policy formulation was examined. RESULTS: The implementation of the screening tests does not take the form of structured mass screening programs. Almost all physicians (urologists-pathologists, gynecologists) apply PSA and routine ultrasonography in normal pregnancy respectively with the purpose of either prevention or diagnosis. Mammography is applied generally for prevention or diagnosis, but there are some mass screening programs at a local level. In addition, the results show no evidence that the efficacy and the cost effectiveness of the three screening programs have been a matter of serious concern and investigation for the purposes of policy formulation in Greece. CONCLUSION: The results point to a need for the implementation of HTA methods on mass screening preventive programs in which real value and cost remain unclear and whose use is based on empirical and personal assessments. PMID- 11495379 TI - Mammography, routine ultrasonography in pregnancy, and PSA screenings in Italy. AB - OBJECTIVE: To present current scientific statements and practice on mammography, ultrasonography in pregnancy, and PSA within the Italian National Health Service (NHS). METHODS: Information on the three screenings was found in several position papers prepared by working groups at national and regional levels. RESULTS: For mammography the adjusted mortality rate for breast cancer increased in the last four decades and decreased in the last few years. Only 4.9% of women aged 50-70 years are covered by screening. In 1996 a consensus paper stated that biennial examination for women aged 50-69 years should be offered to reduce breast cancer mortality. A national program for breast cancer screening was launched under the responsibility of regional governments, but data about its implementation are not yet available. Regarding PSA screening, the results show that prostate cancer is the fourth most frequent neoplasm in males. A formal evaluation of prostate cancer screening was done in 1995 by the National Research Council and the Italian Association for Cancer Research; they did not recommend the NHS implement this service. In the National Health Plan 1998-2000, prostate cancer screening is not mentioned. So far, an organized and standardized screening program, based on routine PSA testing, does not exist in Italy. Nevertheless, a lot of PSA tests are performed at the clinical level to detect early prostate cancers. The results with regard to routine ultrasonography in pregnancy show that in 1996 the birth rate was 9.30 per 1,000 inhabitants, with 536,740 live births. In the period from 1992-94, the percentage of babies with a birth weight less than 2,500 gm and less than 1,500 gm were 5.85% and 0.91%, respectively. A formal screening program for detecting fetal growth retardation, based on the routine ultrasonography in the third trimester of the normal pregnancy, doesn't exist in Italy, but this test is widely performed. It is paid for under NHS coverage. CONCLUSION: In 1998 the National Health Plan 1998-2000 stated that the NHS should offer only evidence based preventive and diagnostic interventions and stressed the importance of appropriate use of health technology. PMID- 11495380 TI - Health technology assessment and screening in The Netherlands: case studies of mammography in breast cancer, PSA screening in prostate cancer, and ultrasound in normal pregnancy. AB - OBJECTIVE: To review the assessment and implementation of three screening methods: mammography for breast cancer, screening for prostate cancer, and routine use of ultrasound in pregnancy. METHODS: To review policy documents and published papers dealing with prevention and screening in the Netherlands, focusing on the three screening methods specified. RESULTS: The results indicate that the Netherlands has an active establishment devoted to health technology assessment (HTA). The Netherlands government has also made prevention a high priority in the health services system. Within prevention policy, HTA is given an important place. The general policy is that prevention programs should meet high standards of effectiveness and efficiency, as well as ethical, legal, and social acceptability. In addition, the Netherlands may be unique in the world in having a specific law requiring that proposals for population screening must be carefully assessed before they are implemented. CONCLUSIONS: The three cases examined in this paper have all been assessed, and the conclusions are similar to those presented in the synthesis published in this issue (33). In the case of mammography, the assessment was followed by a rational implementation of a national screening program for breast cancer. In the other two cases, however, despite negative conclusions from assessment, the tests are frequently carried out, especially in what has been termed opportunistic screening. Prostate cancer screening seems to be spreading rapidly. Use of ultrasound in pregnancy is frequent, not necessarily for medical reasons but because parents wish to have a picture of their fetus. The conclusion is that HTA is well established in the Netherlands, as illustrated in these three cases, and policy is based on the assessments done. However, practice is not in accord with the assessment in the cases of prostate cancer and routine ultrasound. Policies to deal effectively with opportunistic screening are difficult to imagine. PMID- 11495381 TI - Health technology assessment and screening in Sweden. AB - OBJECTIVES: To describe health technology assessment (HTA) and policies concerning three screening procedures in Sweden. METHODS: The main source of information was reports from the Swedish Council for Technology Assessment in Health Care (SBU) and other governmental reports, supplemented by the professional literature. RESULTS: Prevention is emphasized in the healthcare services of Sweden. Specifically, screening is encouraged and supported when it is deemed beneficial. Sweden has a strong orientation toward evidence-based health care and HTA. Since its inauguration in 1987, SBU has fostered the use of HTA in making policy and clinical decisions in Sweden. Government policy in Sweden is to encourage services that are beneficial and cost-effective and discourages services that are not. Screening is no exception to this general rule. The three cases examined in this paper-mammography screening, PSA screening, and routine ultrasound screening in pregnancy-have all been formally assessed in Sweden. Assessments have been an integral part of policy making concerning these and other preventive measures. Mammography screening has been widely implemented. However, as in other countries, screening is often carried out in an opportunistic fashion, so that PSA screening, in particular, is carried out more in Sweden than can be justified by the evidence. CONCLUSIONS: Mammography screening is promoted and is completely available to the target group. PSA screening is discouraged, but not with complete success. Ultrasound in pregnancy is widely used, not because of good evidence of impact on mortality and morbidity among newborns, but because it increases the detection rate of congenitally malformed fetuses and because of evidence of positive effects on the management and planning of deliveries, as well as because of psychological and ethical implications of the technology. HTA is an important part of health policy making in Sweden. PMID- 11495382 TI - Health technology assessment of three screening methods in Switzerland. AB - OBJECTIVE: To describe the level of health technology assessments for three screening methods in Switzerland. METHODS: Analysis of documents and expert opinions on mammography screening, ultrasound examinations during normal pregnancy, and screening for prostate-specific antigen (PSA) with a focus on services provided, recommendations, formal regulations, national papers, projects, and formal assessments. RESULTS: Preventive services are explicitly recommended by the Swiss federal law on health insurance. Two routine ultrasound examinations during normal pregnancy as well as PSA analysis for men over 40 years of age on medical indication are covered by basic health insurance. Mammography screening every 2 years has been covered since 1997 for women over 50 years. A systematic screening program for mammography exists in the western part of Switzerland, and a nationwide program is planned. However, a national program may not come into being until a quality assurance program for evaluation is established, and a nationwide fee for reimbursement-to include all costs of such a program-is accepted by providers and insurers. CONCLUSION: According to the Swiss health insurance law, the effectiveness of mammography screening and ultrasound examinations during normal pregnancy have to be proven. Systematic evaluation of these screening methods is in preparation. PSA is not part of current evaluation. It can be concluded that health technology assessment in Switzerland is now required by law for several medical services. However, limited financial and personnel resources as well as the lack of disease registers may hamper progress in the near future. PMID- 11495383 TI - Evidence-based screening in the United Kingdom. AB - OBJECTIVE: To review the assessment of screening in the United Kingdom, focusing on three methods: mammography for breast cancer, screening for prostate cancer, and routine use of ultrasound in pregnancy. METHOD: To review policy documents and published papers dealing with prevention and screening in the United Kingdom. RESULTS: Indicate that the United Kingdom has an active policy concerning the assessment of screening methods. Generally speaking, this assessment policy is part of the national program for health technology assessment (HTA). The government has given HTA an important place within health care in the United Kingdom, and prevention and screening is no exception to this general rule. The assessment of screening is now implemented through the National Screening Committee, established in 1997. The three issues reviewed in this paper have all been assessed within the context of the Department of Health. In the case of mammography, the assessment was done more than 10 years ago and was followed by a rational implementation of a national screening program for breast cancer. In the case of prostate cancer screening, two systematic reviews have concluded that screening should not be carried out. In general, this recommendation has been accepted in the United Kingdom. Use of ultrasound in pregnancy has been assessed by the National Screening Committee. This complex technology is difficult to assess, and the screening procedure is deeply embedded in clinical practice in the United Kingdom, so assessment has not had much impact on the frequency of screening. CONCLUSION: HTA and the assessment of screening are well established in the United Kingdom. Policy is generally based on the assessments done, and practice generally follows the results of assessment. Assessment of screening is expected to become increasingly important in the United Kingdom during the next years. PMID- 11495384 TI - Health policy, health technology assessment, and screening in Europe. AB - OBJECTIVE: To present a summary of the papers in this volume, illustrating the links between health technology assessment (HTA), health policies and specifically policies toward prevention and screening, and implementation of screening tests in the case of three screening methods: mammography screening for breast cancer, screening for prostate cancer, and routine use of ultrasound in pregnancy. METHOD: To commission papers from eight European countries based on a set of questions to be answered, as well as a paper synthesizing the scientific literature on the three screening procedures. RESULTS: Indicate that few of the countries examined have developed effective links between HTA, health policy, and implementation related to screening procedures. Only Sweden and the Netherlands appear to have developed such links. In the other countries, HTA has a limited role in determining health policy. It also seems to be uncommon for countries to have a specific prevention strategy. CONCLUSION: The major conclusion is that countries of Europe need to develop HTA as part of prevention policies. PMID- 11495385 TI - Single-market regulation and innovation in Europe's medical devices industry. AB - In this article we analyze the influence of the legal regulatory framework in Europe, established by the two directives on medical devices and active implantable devices, on the performance of innovation in a single European market. First, we describe in general the possible influence of a single European market on innovation and the institutional features of the particular harmonization approach ("New Approach") we are looking at here. The empirical results presented derive from a survey investigation involving 150 firms that we defined as best innovators in the European medical devices industry from a pre survey. The results confirm that the total impact of the New Approach regulation on firms' innovation in the long term is positive. However, it also becomes clear that the impact of regulation on innovation is limited if the factors are looked at individually and that there is a clear difference regarding short-term effects. To improve the regulatory framework, several policy actions are recommended. PMID- 11495386 TI - A method for research programming in the field of evidence-based medicine. AB - OBJECTIVE: This paper presents the method we used in programming research on the efficacy of therapeutic interventions for nine chronic benign pain disorders. METHODS: We started with an inventory to identify commonly applied interventions. For these interventions we searched the literature to identify gaps in evidence. First, we searched for recent reviews, of which we assessed the methodologic quality. If only reviews of poor or moderate quality were found, a new systematic review was recommended. When we found no recent reviews, we searched for randomized controlled trials (RCTs). If there was insufficient or inconsistent evidence, the performance of a new RCT was advised. This structured procedure resulted in a list with topics for which new systematic reviews were recommended and topics for which new RCTs were needed. These lists were the starting points for priority setting by four Centers for Pain Management and Research. All members of the centers were asked to state the priority of each topic. The resulting hierarchy of topics for their own center was discussed in a meeting in each center, giving participants the opportunity to elucidate their views and considerations. RESULTS: The final result was a robust priority list for the need of research (systematic reviews and RCTs) on chronic benign pain syndromes. DISCUSSION: The strength and weaknesses of this approach are discussed. This method of priority setting is by no means restricted to treatments for chronic pain; it is also a useful approach for programming research to enable evidence based medicine in other fields of interest. PMID- 11495387 TI - Economic evaluation of a nursing-led intermediate care unit. AB - OBJECTIVES: The aim of this paper is to examine the costs of introducing a nursing-led ward program together with examining the impact this may have on patients' outcomes. METHODS: The study had a sample size of 177 patients with a mean age of 77, and randomized to either a treatment group (care on a nursing-led ward, n = 97) or a control group (standard care usually on a consultant-led acute ward, n = 80). Resource use data including length of stay, tests and investigations performed, and multidisciplinary involvement in care were collected. RESULTS: There were no significant differences in outcome between the two groups. The inpatient costs for the treatment group were significantly higher, due to the longer length of stay in this group. However, the postdischarge costs were significantly lower for the treatment group. CONCLUSIONS: The provision of nursing-led intermediate care units has been proposed as a solution to inappropriate use of acute medical wards by patients who require additional nursing rather than medical care. Whether the treatment group is ultimately cost-additive is dependent on how long reductions in postdischarge resource use are maintained. PMID- 11495388 TI - Cost-effectiveness of various strategies for detecting cases of colon cancer. PMID- 11495389 TI - Harbinger of hope or commodity fetishism: "re-cognizing" dementia in an age of therapeutic agents. PMID- 11495390 TI - Wandering in dementia: a longitudinal study. AB - OBJECTIVE: This article analyzes the natural history of wandering behavior throughout the course of dementia. DESIGN: Prospective, 10-year, longitudinal study of wandering behavior in dementia, with autopsy follow-up. SETTING: Participants with dementia, living at home with a carer. All lived in Oxfordshire, UK. PARTICIPANTS: Eighty-six people with dementia who were living at home with a carer and who were able to walk unaided at entry to study. MEASURES: At 4-monthly intervals, the carers were interviewed using the Present Behavioural Examination to assess wandering behavior in detail; participants with dementia were assessed cognitively. Nine types of "wandering" behavior were distinguished. RESULTS: Changes in wandering behavior were not generally related to gender, age, or time since onset of dementia. Onset of different types of wandering behavior showed some relationship with cognitive state. Various forms of increased walking first appeared during moderate dementia, each type typically persisting for 1 to 2 years. Late dementia was characterized by decreased walking and immobility. CONCLUSIONS: Wandering behavior in dementia can cause great problems for carers. There are different causes for such changes, some of which are related to cognitive ability, for example increased confusion results in ineffectual "pottering" and getting lost. Increased walking at night corresponds with disruption of diurnal rhythm. PMID- 11495391 TI - Combined magnetic resonance imaging and single-photon emission tomography scanning in the discrimination of Alzheimer's disease from age-matched controls. AB - OBJECTIVE: To compare the utility of temporal lobe magnetic resonance imaging (MRI) and single-photon emission tomography (SPET) scanning in discriminating between subjects with Alzheimer's disease (AD) and age-matched controls. METHODS: Thirty subjects with NINCDS-ADRDA AD (23 probable AD, 5 possible AD, 2 definite AD) and 22 age- and sex-matched controls underwent T1-weighted coronal MRI scanning (0.3 T) and technetium 99m-HMPAO SPET scanning. MRI scans were analyzed using a digitizer system with volumes of hippocampus, amygdala, entorhinal cortex, parahippocampal gyrus, and whole cerebral cortex calculated. From SPET scans, regional cerebral blood flow (rCBF) was assessed in anterior and posterior frontal, parietal, occipital, and mesial temporal cortex using a region of interest analysis with the cerebellum as a reference area. RESULTS: Using MRI, the areas that best separated groups were left hippocampal and left amygdala volume, resulting in correct classification (patient vs. control) in 79% of cases (sensitivity 77%, specificity 82%). Exactly the same proportion of subjects were correctly classified by SPET, with the most discriminating rCBF changes being left parietal and right posterior frontal. Combining information from both scans improved the proportion of correctly classified subjects in a discriminant function to 90% (sensitivity 93%, specificity 86%; only 2 AD and 3 controls misclassified). All AD subjects had abnormalities on MRI and/or SPET (sensitivity for combined examinations 100%), while abnormalities on both MRI and SPET had a positive predictive value of 100% for dementia (including the detection of one control subject who later had dementia). Significant correlations between MRI and SPET measures were seen in control subjects but not in patients. CONCLUSION: Both 0.3 T MRI and single rotating gamma camera SPET were equally useful in separating AD subjects from age-matched controls, although the combination of both significantly enhanced discrimination. In particular, all AD subjects had abnormalities on either MRI or SPET and both techniques may have an important role in assisting with clinical diagnosis, though replication in other centers and examination of differentiation of AD from other causes of dementia need to be examined. PMID- 11495392 TI - The Alzheimer's disease activities of daily living international scale (ADL-IS). AB - BACKGROUND: Activities of daily living (ADL) deficits are integral components of dementia disorders, and ADL measures are among the most robust markers of the course of Alzheimer's disease (AD). Despite this acknowledged importance, no clearly useful ADL instrument for cross-cultural application in pharmacologic trials in the early stages of AD had been available. METHOD: An international effort was launched to develop an ADL scale for pharmacologic trials in early AD. Steps taken from 1990 to the present included: (1) international scientific working group meetings and reviews, (2) reviews of existing measures, (3) collating of existent, nonredundant items, (4) querying experts for new items, (5) interviews with informants and subjects in the USA, France, and Germany, toward the identification of potential new items, (6) identification of an item pool based upon these procedures, (7) creation of a trial instrument, (8) piloting of this instrument, and (9) refinement of the scale based upon statistical analysis of the pilot data. Final item selection was based upon: (1) relevance for > or = 80% of subjects in severity-stratified USA and German samples; (2) absence of gender and national biases; (3) significant (p <.05) discrimination between (a) normal versus mildly impaired and (b) mildly impaired versus moderately to moderately severely impaired subjects; and (4) Global Deterioration Scale (GDS) scores accounting for > or = 12% of variance in the item after controlling for age and gender. RESULTS: An ADL scale consisting of 40 items that correlate with the global and cognitive progress of AD is developed for international usage in pharmacologic trials in incipient, mild, moderate, and moderately severe AD. The scale contains 40 items falling within 13 ADL categories. The 40-item scale is shown to have .81 correlation with GDS staging, .81 with mental status assessment (Mini-Mental State Examination), and .81 with a psychometric test (the SKT) (p values < .001). CONCLUSION: This scale can be used to measure therapeutic response in AD. PMID- 11495393 TI - Agitation in demented patients in an acute care hospital: prevalence, disruptiveness, and staff burden. AB - BACKGROUND/LITERATURE REVIEW: The prevalence of agitated behaviors in different populations with dementia is between 24% and 98%. Although agitated behaviors are potentially disruptive, little research attention has been focused on the effects of these behaviors upon nursing staff. The objectives of this study of demented patients in long-term-care beds at an acute care community hospital were to determine the frequency and disruptiveness of agitated behaviors; to investigate the associations of patient characteristics and interventions with the level of agitation; and to explore the burden of these agitated behaviors on nursing staff. METHOD: The study sample comprised 56 demented patients in the long-term care unit during the study period. Twenty-seven staff who cared for these patients during three shifts over a 2-week period were interviewed to rate the frequency and disruptiveness of agitated behaviors using the Cohen-Mansfield Agitation Inventory, and the burden of care using a modified version of the Zarit Burden Interview. Data on patient characteristics and interventions extracted from the hospital chart included scores on the Barthel Index and Mini-Mental State Examination, the use of psychotropic medication, and the use of physical restraints. RESULTS: Ninety-five percent of the patients with dementia were reported to have at least one agitated behavior; 75% had at least one moderately disruptive behavior. A small group of six patients (11%) had 17 or more disruptive behaviors. The frequency of most behaviors did not vary significantly by shift. Length of stay on long-term care, Barthel Index score, and the use of psychotropic medications were significantly associated with the number of agitated behaviors. The number of behaviors, their mean frequency, and their mean disruptiveness were all significantly correlated with staff burden. DISCUSSION: The prevalence of agitated behaviors in patients with dementia in long-term-care beds at an acute care hospital is similar to that reported in long-term-care facilities. These behaviors are associated with staff burden. PMID- 11495394 TI - Long-Term use of rivastigmine in patients with dementia with Lewy bodies: an open label trial. AB - Patients with dementia with Lewy bodies (DLB) have progressive deficits in cognition, parkinsonism, and neuropsychiatric symptoms. Cholinesterase inhibitors have been used to ameliorate cognitive decline and neuropsychiatric symptoms in short-term trials. In this study, patients with DLB were treated with rivastigmine up to 96 weeks. Improvement from baseline was seen in cognitive function as measured by the Mini-Mental State Examination (MMSE), and neuropsychiatric symptoms as measured by the Neuropsychiatric Inventory (NPI) over the first 24 weeks of treatment. By 96 weeks, neither the MMSE scores nor the NPI scores were significantly worse than at baseline. PMID- 11495395 TI - Alcohol, aging, and cognitive performance in a cohort of Japanese Americans aged 65 and older: the Kame project. AB - OBJECTIVE: To investigate the effects of light to moderate alcohol consumption on cognitive performance. DESIGN AND SETTING: A cross-sectional analysis including older Japanese Americans in King County, WA, enrolled in the Kame Project, a population-based study of cognition, dementia, and aging. PARTICIPANTS: 1,836 cognitively intact participants aged 65 and older who participated in the baseline (1992-1994) examination. MEASUREMENT: Cognitive performance was measured using the Cognitive Abilities Screening Instrument, reaction time (simple and choice), and a measure of vocabulary (North American Adult Reading Test). RESULTS: Multivariate analyses were used to examine the relationship between cognitive performance and alcohol consumption at baseline with men and women together and then separately controlling for age, education, smoking, history of stroke, angina, hypertension, diabetes, and coronary heart disease. Findings showed lower cognitive test scores were observed for men who were either abstainers or in the heavy drinking group. For women, a linear relationship between alcohol consumption and cognitive performance was seen on two of the four measures of cognitive functioning. No significant difference in the association of drinking and cognitive function was identified within the different Japanese American subgroups. CONCLUSION: RESULTS suggest a possible positive relationship between light to moderate drinking and cognitive performance in an aging Japanese American population. Additional long-term prospective and cross-cultural studies are needed to determine the generalizability of these findings to other aging cohorts. PMID- 11495396 TI - A controlled study of repetitive transcranial magnetic stimulation as a treatment of depression in the elderly. AB - Rapid transcranial magnetic stimulation (rTMS) applied to the left dorsal lateral frontal cortex has been shown to produce antidepressant effects. Older depressed patients, however, in one study showed a lower response rate than younger patients. The current study examined treatment response in 20 depressed, treatment-refractory patients (mean age 60.7 +/- 9.8 years) given five sessions of rTMS at 20 Hz for 2 seconds over 20 trains at 80% of motor threshold or identical placebo stimulation, after patients had been withdrawn from their antidepressants. There were no significant differences in Hamilton Depression Scale scores either before or after treatment at 7 days' follow-up. There were three responders to active treatment and three to sham treatment and responders had significantly greater frontal lobe volume than nonresponders (p = .03). These findings suggest that the stimulation parameters used in this study were probably insufficient to produce treatment response and that frontal atrophy may interfere with the effectiveness of rTMS. PMID- 11495397 TI - A double-blind, placebo-controlled study of fluoxetine in depressed patients with Alzheimer's disease. AB - OBJECTIVE: To examine the efficacy of fluoxetine in the treatment of depression in patients with probable Alzheimer's disease (AD). METHODS: This double-blind, parallel-design study included a consecutive series of 41 AD subjects meeting DSM IV criteria for major or minor depression who were randomized to receive fluoxetine (up to 40 mg/day) or identical-appearing placebo. All patients received biweekly evaluations consisting of the Hamilton Depression Scale (HAM-D) and the Clinical Global Impression as primary efficacy measures, and the Mini Mental State Exam, Hamilton Rating Scale for Anxiety, and the Functional Independence Measure as secondary efficacy measures. RESULTS: Complete remission of depression was found in 47% of subjects treated with fluoxetine and in 33% of subjects treated with placebo. Both the fluoxetine and the placebo groups showed a significant decline in HAM-D scores over time, but the magnitude of mood improvement was similar for both groups. Fluoxetine was well tolerated, and most side effects were mild. CONCLUSION: Fluoxetine treatment for depression in AD did not differ significantly from treatment with placebo. Our study also confirms the presence of a placebo effect in the treatment of depression in AD. PMID- 11495398 TI - Caregiver report of hallucinations and paranoid delusions in elders aged 70 or older. AB - This study examined the demographic, medical, and psychiatric correlates of hallucinations and paranoid delusions reported by proxy informants for 822 elders aged 70 or older. This sample comprised people who were deemed unable to complete a direct interview in a large nationwide study of aging. Marital status, trouble with vision, and cognitive impairment were associated with report of both paranoid delusions and hallucinations. Depressive symptoms and stroke were associated with hallucinations only. These results suggest that inadequate external stimulation in the elderly leads to psychotic experiences. PMID- 11495399 TI - Sleep dentistry-should oral sedation for adults now be regulated? PMID- 11495400 TI - Comparative study on anesthetic potency depending on concentrations of lidocaine and epinephrine: assessment of dental local anesthetics using the jaw-opening reflex. AB - Anesthetic potency of a local anesthetic on the dental pulp was investigated by increasing or decreasing the concentration of lidocaine and that of epinephrine. An electromyogram of the digastric muscle in Japan White male rabbits was recorded during the jaw-opening reflex induced by electrical stimulation of the dental pulp. Probit analysis was used for the determination of the 50% effective volume (ED50) values of the anesthetic. The anesthetics used were plain 2% lidocaine solution (2Lid-0 group), 2% lidocaine solution with 12.5 microgram/mL of epinephrine (2Lid-1/8 group), 2% lidocaine solution with 6.25 microgram/mL of epinephrine (2Lid-1/16 group), and 4% lidocaine solution with 5 microgram/mL of epinephrine (4Lid-1/20 group). No anesthetic effect was shown in the 2Lid-0 group. The 2Lid-1/8 group indicated adequate anesthetic potency with the smallest dosage at all observation periods. The potency in the 2Lid-1/16 group was 0.3-0.5 times, and that in the 4Lid-1/20 group was 0.3-0.4 times as much as the 2Lid-1/8 group. The decrease in epinephrine concentration produced the decrease in the anesthetic potency on the dental pulp independent of lidocaine concentration. These results suggest that the increase in lidocaine concentration may not compensate the decrease in epinephrine concentration. PMID- 11495401 TI - Allergic response to metabisulfite in lidocaine anesthetic solution. AB - True allergies to local anesthetics are rare. It is common for practitioners to misdiagnose a serious adverse event to local anesthetics as an allergic reaction. The most likely causes for an allergic response are the preservative, antioxidant, or metabolites and not the anesthetic itself. This case report illustrates the need for practitioners to understand the many potential allergens in local anesthetics and to correctly diagnose patients that are truly allergic to the local anesthetic. PMID- 11495402 TI - Pneumothorax with soft tissue emphysema following abrupt wake-up and self extubation. PMID- 11495403 TI - Recruiting phobic research subjects: effectiveness and cost. AB - Efficiently enrolling subjects is one of the most important and difficult aspects of a clinical trial. This prospective study evaluated strategies used in the recruitment of 144 dental injection phobics for a clinical trial evaluating the effectiveness of combining alprazolam with exposure therapy. Three types of recruitment strategies were evaluated: paid advertising, free publicity, and professional referral. Sixty-three percent of subjects were enrolled using paid advertising (the majority of them from bus advertisements [27.0%], posters on the University of Washington campus [20.1%], and newspaper advertisements [13.2%]). Free publicity (eg, television coverage, word of mouth) yielded 18.8% of enrolled subjects and professionaL referrals 14.6% of subjects. The average cost (1996 dollars) of enrolling 1 subject was $79. Bus and poster advertising attracted more initial contacts and yielded the greatest enrollment. PMID- 11495404 TI - The use of EMLA for an intraoral soft-tissue biopsy in a needle phobic: a case report. AB - A case is reported of the removal of a leaf fibroma from the mucosa of the hard palate using EMLA topical anesthesia as the sole means of pain control. PMID- 11495405 TI - Anesthetic efficacy of a combination of hyaluronidase and lidocaine with epinephrine in inferior alveolar nerve blocks. AB - The purpose of this prospective, randomized, double-blind study was to determine the anesthetic efficacy of a buffered lidocaine with epinephrine solution compared to a combination buffered lidocaine with epinephrine plus hyaluronidase solution in inferior alveolar nerve blocks. Thirty subjects randomly received an inferior alveolar nerve block using 1 of the 2 solutions at 2 separate appointments using a repeated-measures design. Mandibular anterior and posterior teeth were blindly pulp tested at 4-minute cycles for 60 minutes postinjection. No response from the subject to the maximum output (80 reading) of the pulp tester was used as the criterion for pulpal anesthesia. Anesthesia was considered successful when 2 consecutive readings of 80 were obtained. A postoperative survey was used to measure pain and trismus. The results demonstrated 100% of the subjects had profound lip numbness with both solutions for inferior alveolar nerve blocks. The anesthetic success rates for individual teeth ranged from 20 to 80%. There were no significant differences (P > .05) between the 2 solutions. However, the combination lidocaine/hyaluronidase solution resulted in a significant increase in postoperative pain and trismus. It was concluded that adding hyaluronidase to a buffered lidocaine solution with epinephrine did not statistically increase the incidence of pulpal anesthesia in inferior alveolar nerve blocks and, because of its potential tissue damaging effect, it should not be added to local anesthetic solutions for inferior alveolar nerve blocks. PMID- 11495406 TI - An analysis of spontaneous hatching in a human endometrial epithelial coculture system: is assisted hatching justified? AB - PURPOSE: To evaluate spontaneous embryo hatching in an endometrial epithelial coculture system, and compare it with cases where coculture was performed because of maternal age, previous repeated implantation failures, or both. To clarify in which cases assisted hatching would be appropriate. METHODS: Individual human embryos were cocultured on an endometrial epithelial cell monolayer until Day 6. RESULTS: Blastocyst hatching rate at Day 6, depending on maternal age, was 9.1% (age <37 years) and 3.4% (age > or = 37 years). However, blastocyst hatching rates depending on number of previous IVF failures were similar. CONCLUSIONS: Maternal age and previous implantation failures are factors affecting the ability of human embryos to reach the blastocyst stage in coculture. However, assisted hatching is not justified in these populations because of the absence of hatching rate differences between blastocysts obtained from these two groups and the control group. PMID- 11495407 TI - Antral follicle counting in predicting the retrieved oocyte number after ovarian hyperstimulation. AB - PURPOSE: To evaluate the antral follicle (AF) counting in predicting the outcome after controlled ovarian hyperstimulation (COH) and IVF-ET. METHODS: Infertile women who accepted the COH and IVF-ET were included prospectively. Day PMID- 11495409 TI - Successful Day 5 embryo transfer and pregnancies resulting after transport of embryos by air for biopsy and genetic analysis. AB - PURPOSE: Case studies of four in vitro fertilization (IVF) cycles where embryo transport by commercial airline followed by biopsy and genetic analysis with subsequent culture to Day 5 and resulting ongoing pregnancy. METHOD: Retrospective clinical case study of 4 patients requiring preimplantation genetic diagnosis (PGD) testing. Normally fertilized embryos were transported in a battery-powered portable incubator by commercial airline following evaluation for fertilization under controlled conditions from the Center for Assisted Reproduction, Bedford, Texas to the Reproductive Genetic Institute, Chicago, Illinois. Following Day 3 embryo biopsy and genetic analysis, embryos were transported back to the Center for Assisted Reproduction for Day 5 embryo transfer. RESULTS: Ongoing clinical pregnancy resulted for all patients receiving embryo transfer. CONCLUSION: These results demonstrate the feasibility of embryo transport by air for centers that do not have the in-house capabilities to perform genetic analysis. With successful pregnancies obtained through extended culture to Day 5, embryos requiring genetic analysis can be successfully transported by air, tested, and returned to the initial facility for embryo transfer without time restriction. PMID- 11495408 TI - Pregnancies resulting from in vitro matured oocytes collected from women with regular menstrual cycle. AB - PURPOSE: To demonstrate that human immature oocytes retrieved from women with regular menstrual cycles can undergo maturation and fertilization, and that the resulting embryos can establish pregnancies. METHODS: Immature oocytes (n = 568) were retrieved from women with regular menstrual cycle. The intact immature oocytes (n = 506) were allowed to mature in YS medium supplemented with 70% human follicular fluid (hFF); the matured oocytes were fertilized with husband sperm. Two pronuclei oocytes were cocultured with cumulus cells in YS medium supplemented with 10% hFF until 2 or 3 days after insemination. The cleaved embryos were transferred in uteri. RESULTS: Follicles were aspirated on Day 9.2 +/- 5.3 of 63 natural cycles from 51 patients (mean age = 34.8 +/- 4.0 years). The average number of retrieved immature oocytes was 9.0. The maturation rate was 74.3% (376/506). The two PN and cleavage rates were 72.6% (273/376) and 89.0% (243/273), respectively. Embryo transfer was achieved in 51 cycles and clinical pregnancy rate was 17.6% (9/51). CONCLUSIONS: The results suggest that in vitro matured oocytes can undergo fertilization and the resulting embryos may successfully lead to pregnancies. However, further research is needed to improve IVM technique to achieve success rate comparable to gonadotrophin stimulated cycles. PMID- 11495410 TI - Maternal serum C-reactive protein level does not change significantly after fetal reduction: it could be used as an indicator of chorioamnionitis. AB - PURPOSE: This study was aimed at investigating the diagnostic value of maternal serum C-reactive protein (CRP) in the recognition of chorioamnionitis in patients undergoing fetal reduction. METHODS: Seventy-one gravidas with high-order multifetal pregnancies, including 46 with triplets, 18 with quadruplets, and 7 with quintuplets, who underwent transabdominal fetal reduction to twins during the 10th-14th gestational week were recruited. The subjects were followed up clinically and ultrasonographically 1 week and 1 month after fetal reduction for signs of infection, premature uterine contraction, and premature rupture of the membranes CRP levels were measured prior to fetal reduction and at follow-up examinations, and were compared. RESULTS: Among the 71 mothers, 65 (92%) were normal after fetal reduction. The CRP levels were not significantly different prior to the procedure (0.27 +/- 0.26 mg/dL), and 1 week (0.23 +/- 0.24 mg/dL) and 1 month (0.24 +/- 0.20 mg/dL) later. There was no correlation between the number of fetuses reduced and the CRP levels. Six (8%) experienced leakage of amniotic fluid after fetal reduction. Three patients had normal CRP levels at that time and at the following tests. The pregnancies continued smoothly after conservative treatment. The other three patients had elevated CRP levels when leakage of amniotic fluid occurred. Fever and uterine irritability developed subsequently despite parenteral antibiotics and tocolytic therapy. Daily checks showed increasing CRP levels. The pregnancies were aborted, and the histology of the placental membranes revealed chorioamnionitis with infiltration of acute inflammatory cells. CONCLUSIONS: The absorption of inactive gestational tissue after fetal reduction did not affect CRP levels. CRP may be used as a marker of intrauterine infection after fetal reduction. PMID- 11495411 TI - In vivo hatching phenomenon of mouse blastocysts during implantation. AB - PURPOSE: To determine whether the blastocyst zona shedding process within the murine uterine cornus in vivo is due to a global lytic process caused by uterine proteolytic enzyme, or is triggered by the blastocyst hatching process as observed in vitro. METHODS: Fifty-one female ICR mice aged 5-8 weeks were used for this study. From 8:00 p.m. of the 4th day postcoitus to 7:00 p.m. of the 5th day postcoitus, the uterine cornua of 51 mice were isolated at 30-min intervals. Blastocysts within the uterine cornua were flushed out with a balanced solution under the dissecting microscope. The stages of blastocyst development and the proportion of hatching or hatched blastocysts and the discarded zona pellucida (ZP) were inspected and counted. RESULTS: A total of 672 blastocysts were recovered from the uterine horns of the 51 mice. They were divided into six groups according to the blastocyst developmental stages (before or after ZP escape; before or after the initiation of implantation). Group I represents the earliest embryonic stage and Group VI represents the most advanced blastocyst developmental stage during the peri-implantation period. The empty ZP recovery rates (number of discarded ZP/all hatched blastocysts) were 52.3%, 21.5%, 17.2%, 6.6%, 1.6%, and 0% in Groups I-VI, respectively. Five hatching blastocysts out of 199 embryos in Group I were found and a 100% of ZP recovery rate was obtained from 6 of 19 mice in Groups I and II. CONCLUSIONS: The present study shows that active blastocyst hatching occurs in vivo because both hatching blastocysts and empty ZP can be found within the uterine cornua of ICR mice before implantation. The empty ZP recovery rate declined significantly, along with a progression of embryo development and implantation, implying that intrauterine zona lytic activity occurs during the peri-implantation stage. PMID- 11495412 TI - Birth of a healthy infant after in vitro fertilization and embryo transfer in patient of total uterine prolapse. PMID- 11495413 TI - Combined intrauterine, tubal, and cervical pregnancies following in vitro fertilization and embryo transfer. PMID- 11495414 TI - Failure to recover oocytes during IVF: case reports and suggestions for management. PMID- 11495415 TI - Ovarian hyperstimulation after the sole use of a gonadotrophin-releasing hormone agonist (Nafarelin) as a complication of in vitro fertilisation treatment. PMID- 11495416 TI - Ottorino Rossi Award 2001. Mitochondrial DNA: a genetic Pandora's box. PMID- 11495417 TI - Neutralizing antibodies (NABS) and interferon beta-1b therapy of multiple sclerosis. PMID- 11495418 TI - The trigemino-cervical reflex in normal subjects. AB - There are only few reports on the trigemino-cervical reflex in humans and there is debate over the best method of reflex examination. The aim of this study was, comparing different methods, to provide a reproducible method for evaluating the trigemino-cervical reflex. The trigemino-cervical reflex was studied in 32 healthy volunteers. The stimulation was applied to the supraorbital, infraorbital or mental nerve. Recordings were performed bilaterally from the sternocleidomastoid and trapezius muscles at rest. The reflex was also examined during maximal voluntary contraction of the sternocleidomastoid muscle after supraorbital nerve stimulation. It presented as a two-component reflex if recorded from a tonically active muscle and as a one-component reflex if recorded from a relaxed muscle. The most reproducible reflex responses were obtained from the resting sternocleidomastoid muscle after stimulation of the supraorbital nerve. In conclusion, the trigemino-cervical reflex may be most easily obtained from the relaxed sternocleidomastoid muscle after supraorbital nerve stimulation. PMID- 11495419 TI - The position of the head in space: a kinematic analysis in patients with cervical dystonia treated with botulinum toxin. AB - Many instruments have been employed in recent years in order to quantify the posture and motion of the head in normal and pathological subjects. Evaluations of this type present many difficulties related to the influence of individual and external factors and to the accuracy of the system used. In patients with cervical dystonia (CD) the only rating scales currently used are semi quantitative and subjective. More precise information on disease severity and response to the treatment is needed. Posture and motion of the head were evaluated by means of ELITE motion analyser (BTS, Milan, Italy) in 6 patients with the left laterocollis form of CD undergoing treatment with botulinum toxin (BTX). The method emerged as very useful for the quantification of the therapeutic response (which was more marked in motion than in posture). We found an inverse relationship between the degree of motion improvement and the restriction of motion before treatment. PMID- 11495420 TI - Parkinsonian rigidity. PMID- 11495421 TI - Rehabilitation of parkinsonian patients. PMID- 11495422 TI - Auditory rhythmicity enhances movement and speech motor control in patients with Parkinson's disease. PMID- 11495423 TI - Solvent-free routes to clean technology. AB - A major aim for the chemical technology of the future is the avoidance of noxious and environmentally unacceptable organic solvents. In this concept article we discuss more environmentally friendly and highly selective alternatives which we have evolved for carrying out a number of important chemical conversions. These entail the use of porous heterogeneous catalysts in which the active sites have been atomically engineered and fully characterized. Such solid catalysts operate under solvent-free conditions and usually entail one-step processes. PMID- 11495424 TI - Novel 4-vinylpyridine-extended metal-dibenzoylmethanate host frameworks: structure, polymorphism, and inclusion properties. AB - In this contribution we show that host materials based on metal dibenzoylmethanates (DBM) can be extended in a versatile way by decreasing the packing efficiency of the simpler metal DBM's reported earlier. Specifically, this can be accomplished by coordinating two 4-vinylpyridines (4-ViPy) to the metal (Ni or Co) DBM units to give [M(4-ViPy)2(DBM)2] host complexes. These display a remarkable polymorphism and an ability to form inclusion compounds with a large variety of organic species. Five non-clathrate phases representing three polymorphic types and twenty-eight inclusion compounds with nineteen guests, representing five structural types were isolated and studied in varying degrees of detail. The inclusion compounds can be prepared by recrystallization or by interaction of the solid host with guest vapor. In the latter case, the process realization, kinetics and final product strongly depend on the host polymorph chosen as starting material. Kinetic studies executed with powder XRD suggest that transient formation of inclusion compounds may occur even during solvent vapor induced transformation of one guest-free polymorph to another. The beta polymorph of the Ni-host reveals the strongest clathratogenic ability as well as a high selectivity towards certain homologues and isomers. Its properties give insight into the concept of "flexible zeolite mimics", or "apohosts", as this empty host form is energetically and structurally predisposed towards inclusion processes. In all eleven (three host and eight clathrate) structures studied by single crystal X-ray diffraction the [M(4-Vi-Py)2(DBM)2] complex molecule is transconfigured. In most, the host molecules show effective packing in one dimension by forming parallel chains. Guest species are located between the chains in cages or channels formed by combining voids in the host molecules belonging to adjacent chains. The corresponding Ni and Co versions of the compounds studied were similar. PMID- 11495425 TI - First experimental observation on different ionic states of the tert-butoxy [(CH3)3CO*] radical. AB - A continuous tert-butoxy (CH3)+CO* radical beam is produced in situ by respective pyrolysis of both (CH3)3CONO at 115(+/-0.5) degrees C and (CH3)3COOC(CH3)3 at 87(+/- 0.5) degrees C. By combining the HeI photoelectron (PE) spectrum with the improved density function theory (DFT) calculations, we have concluded that the (CH3)3CO* radical has C3V symmetry and X2E ground state. The study does not only provide the ionization energies of different ionic states of the (CH3)3CO* radical for the first time, but also the first example in which there have been similar vibrational structures in different ionic states caused by removal of the electron on an orbital. It is also pointed out that (CH3)3CONO is a good source for obtaining the (CH3)3CO* radical beam, and that NO is a stable regent for the active radical. The results will promote the studies in electron spin resonance (ESR) research on the mechanisms of both the initiation of the formation of a new radical and the radical-chain polymerization in which the (CH3)3CO* radical participates. PMID- 11495426 TI - Novel chiral macrocycles containing two electronically interacting arylene chromophores. AB - Novel chiral macrocycles consisting of two rigid oligoarylene rods and two chiral spiroindane clips have been synthesized by condensation of spiroindane diols and CF3-activated alpha-omega-difluorooligoaryls. Since a broad variety of planar aromatic macrocycles is known, our non-planar, chiral rings represent a new class of macrocyclic compounds. The first two examples, which contain quaterphenylene and diphenylbithiophene rods, are presented in this communication; for one of them a crystal structure is given. The chiroptical properties of the macrocycles can be interpreted as an interplay of the "intra-rod" helicity of individual oligoarylene rods and the "inter-rod" helicity between both chromophores of the macrocycle. The macrocycles can act as chiral dopands of commercially available, and novel, polymeric nematic liquid crystals (emissive polyfluorenes). The "intra rod" helicity of individual oligoarylene rods is the main feature in determining the resulting helical twisting power (HTP). The cholestric induction in mesogenic, emissive polyfluorenes is of special interest for a realization of electronic devices that have a circularly polarized electroluminescence. The results are also important for an understanding of larger ensembles of chiral rodlike molecules, especially their pi-pi interactions. PMID- 11495427 TI - Competitive decay pathways of the radical ions formed by photoinduced electron transfer between quinones and 4,4'-dimethoxydiphenylmethane in acetonitrile. AB - The reactivity of the cation radical of (4-MeOC6H4)2CH2 photosensitized by 1,4 benzoquinone (BQ), 2,5-dichloro-1,4-benzoquinone (Cl2BQ), and tetrachloro-1,4 benzoquinone (chloranil, CA) was investigated in acetonitrile. The main photoreaction products obtained by steady-state irradiation were identified to be: (4-MeOC6H4)2-CHOC6H4OH, sensitized by BQ; (4-MeOC6H4)2CHCl, sensitized by Cl2BQ; (4-MeOC6H4)2CHOH, sensitized by CA. The mechanism of their formation was investigated by nanosecond laser flash photolysis that allowed transient species (radical ions, neutral radicals, and ions) to be detected and characterized in terms of absorption spectra, formation quantum yields, and decay rate constants. For all systems, the interaction between the triplet quinone (Q) and (4 MeOC6H4)2CH2 produced the corresponding radical ions (quantum yield phi > or = 0.72) which mainly decay by back electron transfer processes. Less efficient reaction routes for the radical ions Q*- and (4-MeOC6H4)2CH2*+ were also: i) the proton-transfer process with the formation of the radical (4-MeOC6H4)2CH* by use of Cl2BQ; ii) the hydrogen-transfer process with the formation of the cation (4 MeOC6H4)2CH+ in the case of CA. Instead. BQ sensitized a much higher yield of BOH* and (4-MeOC6H4)2CH*, mainly by the direct interaction of triplet BQ with (4 MeOC6H4)2CH2. It was also shown that the presence of salts decreases significantly the rate of the back electron transfer process and enhances the quantum yields of formation of the neutral radicals and ions when Cl2BQ and CA are used, respectively. The behavior of BQ*-, Cl2BQ*-, and CA*- appears to be mainly determined by the Mulliken charges on the oxygen atom obtained from quantum mechanical calculations with the model B3LYP/6-311G(d,p). Spin densities seem to be much less important. PMID- 11495428 TI - High-spin iron(II) as a semitransparent partner for tuning europium(III) luminescence in heterodimetallic d-f complexes. AB - The segmental ligand 2-[6-(N,N-diethylcarbamoyl)pyridin-2-yl]-1,1'-dimethyl-5,5' methylene-2'-(6-methylpyridine-2-yl)bis[1H-benzimidazole] (L3) reacts with a stoichiometric mixture of LnIII (Ln = La, Eu, Gd) and M(II) (M = Zn, Fe) in acetonitrile to produce selectively the heterodimetallic triple-stranded helicates (HHH)-[LnM(L3)3]5+. In these complexes, M(II) is pseudooctahedrally coordinated by the three wrapped bidentate binding units, thus forming a noncovalent tripod which organizes the three unsymmetrical tridentate segments to give ninefold coordination to LnIII. The introduction of a methyl group at the 6 position of the terminal pyridine in L3 sterically reduces the complexing ability of the bidentate segment for M(II). Spectroscopic (ESI-MS, UV/Vis/NIR, NMR), magnetic and electrochemical measurements show that 1) the head-to-head-to-head triple helical complexes (HHH)-[LnM(L3)3]5+ are quantitatively formed in solution only for ligand concentrations larger than 0.01 M, 2) FeII adopts a pure high spin electronic configuration in (HHH)-[LnFe(L3)3]5+ and 3) the FeII/FeIII oxidation process is prevented by steric constraints. Detailed photophysical studies of (HHH)-[Eu-Zn(L3)3]5+ confirm that the pseudotricapped trigonal prismatic lanthanide coordination site is not affected by the methyl groups bound to the terminal pyridine, thus leading to significant Eu-centered emission upon UV irradiation. In (HHH)-[EuFe(L3)3]5+, a resonant intramolecular Eu-->Fe(II)hs energy transfer partially quenches the Eu-centered luminescence; however, the residual red emission demonstrates that high-spin iron(II) is compatible with the sensitization of Eu(III) in heterodimetallic d-f complexes. The influence of the electronic configuration of Fe(II) on the efficiency of Eu(III)-->Fe(II) energy transfer processes is discussed together with its consequence for the design of optically active spin-crossover supramolecular devices. PMID- 11495429 TI - Stannaborate transition metal chemistry: ligand properties, reactivity, and density functional theory calculations of platinum and palladium complexes. AB - Three stannaborate complexes of platinum(II) and a novel stannoborate palladium(II) derivative have been prepared in excellent yield. The tin transition metal bond is formed through nucleophilic substitution and the resulting complexes [Bu3MeN] [trans-[(Et3P)2Pt(SnB11H11)H]] (6), [trans [(Et3P)2Pt(SnB11H11)(CNtBu)]] (7), [Bu3MeN]2[trans-[(Et3P)2Pt(SnB11H11)2 (CNtBu)]] (8), and [Bu3MeN][(dppe)-Pd(SnB11H11)Me] (12) (dppe = 1,2-bis (diphenylphosphanyl)ethane) were characterized by NMR spectroscopy and elemental analysis. In the cases of the zwitterion 7, the pentacoordinated complex 9, the palladium salt 12 and [(triphos)Pt(SnB11H11)] (10) (triphos = 1,1,1 tris(diphenylphosphanylmethyl)ethane), their solid-state structures are determined by X-ray crystal structure analyses. The trans influence of the [SnB11H11] ligand is evaluated from the results of the IR spectroscopy and X-ray crystallographic structures of complexes 6, 7, and 12. The dipole moment of the zwitterion 7 is calculated by density functional theory (DFT) methods. The alignment of the dipole moments of the polar molecules 7 and 12 in the solid state is discussed. PMID- 11495430 TI - Three- and four-membered rings from cycloadditions of 1,3-thiazolium-4-olates and aldehydes. AB - 2-Aminothioisomunchnones, a well-known family of masked dipoles, react with aromatic aldehydes in a domino cascade reaction that produces episulfides (thiiranes) or beta-lactams (2-azetidinones). This sequence is initiated by a [3+2] dipolar cycloaddition followed by ring opening of cycloadducts and intramolecular rearrangement to afford these unusual ring contractions. The nature of the reaction products depends on the structural characteristics of the starting dipole and the experimental conditions. Episulfides are obtained selectively as cis isomers with respect to both aryl groups, whereas beta-lactams are produced as cis/trans mixtures. These structural features were determined unequivocally by X-ray crystallographic analysis. The beta-lactams still possessed a flexible acyclic chain containing sulfur, a salient lead modification of the bioactive cyclic penems and cephems. The preferential production of exo transition structures was rationalized with the aid of computational calculations at the B3LYP/6-31G* level. PMID- 11495431 TI - Reactions of aliphatic ketones R2CO (R=Me, Et, iPr and tBu) with the MCI4/Li(Hg) system (M=U or Ti): mechanistic analogies between the McMurry, Wittig, and Clemmensen reactions. AB - Analysis of the products of the reactions of ketones R2CO (R = Me, Et, iPr, tBu) with the MCl4/Li(Hg) system (M = U, Ti) at 20 degrees C revealed significant differences. For R = Me, the reaction proceeded exclusively (M = U) or preferentially (M = Ti) via a metallopinacol intermediate resulting from dimerization of ketyl radicals. Pinacol was liberated by hydrolysis, and tetramethylethylene was obtained after further reduction at 65 degrees C. For R=iPr, formation of iPr2C=CiPr2 as the only coupling product, the nonproduction of this alkene by reduction of the uranium pinacolate [U]-OCR2CR2O-[U] (R= iPr) at 20 degrees C, and the instability of the corresponding titanium pinacolate towards rupture of the pinacolic C-C bond indicated that reductive coupling of iPr2CO did not proceed by dimerization of ketyl radicals. Formation of 2,4 dimethyl-2-pentene was in favor of a carbenoid intermediate resulting from deoxygenative reduction of the ketyl. These results revealed that for sterically hindered ketones, McMurry reactions can be viewed as Wittig-like olefination reactions. For R=tBu, no coupling product was obtained and the alkane tBu2CH2 was the major product. The involvement of the carbenoid species [M]=CtBu2 was confirmed by its trapping with H2O, leading to tBu2CH2, and with the aldehydes RCHO, giving the cross-coupling products tBu2C=C(R)H (R = Me, tBu). Therefore, in the case of severely congested ketones, McMurry reactions present strong similarities to the Clemmensen reduction of ketones, owing to the involvement in both reactions of carbenoid species which exhibit similar reactivity. PMID- 11495432 TI - Metal-metal and carbon-carbon bonds as potential components of molecular batteries. AB - The reductive coupling of [M(salophen)] derivatives, where M is an early transition metal and salophen is N,N'-o-phenylenebis(salicylideneaminato) dianion, led to the formation of dimers linked through C-C and M-M bonds. Both of these bonds can potentially function as electron reservoirs: each bond can be used as a reversible source of a pair of electrons under the condition that it is not chemically transformed by the incoming substrate which functions as an electron acceptor. To explore this potential function as well as the competition in the redox processes between C-C and M-M bonds within the same molecular framework, we investigated the reduction of [(tBu4-salophen)NbCl3] (1) and [(tBu4 salophen)MoCl2] (7) as model compounds. In the former case, the reduction led to [(Nb-Nb)(tBu4-*salophen2*)] (2) which contains both a Nb-Nb bond (2.6528(7) A) and two C-C bonds across two imino groups of the ligand. Complex 2 can be reduced further to a transient compound 5 that contains an Nb=Nb bond. In the second case, the reduction of 7 by two electrons led to [(Mo[triplebond]Mo)(tBu4 salophen)2] (8), which does not contain any C-C linkages between the two salophen units. Complexes 2 and 5 are able to transfer one pair and two pairs of electrons, respectively, to give compounds 3, 4, and 6, with the consequent cleavage of the Nb-Nb and Nb=Nb bonds. In the present case, it is surprising that the C-C bonds do not participate in the reduction of the substrates. A careful theoretical treatment anticipates, both in the case of 1 and 7, the preferential formation of metal-metal bonds upon reduction. This is indeed the case for 7, but not for 1, where the formation of C-C bonds competes with that of M-M bonds, the latter being the first ones, however, to be involved in electron-transfer reactions. The theoretical approach allowed us to investigate the possibility of intramolecular electron transfer from C-C bonds to M-M bonds and vice versa. PMID- 11495433 TI - Synthesis of (E)-alpha,beta-unsaturated amides with high selectivity by using samarium diiodide. AB - Stereoselective beta-elimination of 2-chloro-3-hydroxyamides 1 is achieved by using samarium diiodide to yield alpha,beta-unsaturated amides 2, in which the C=C bond is di- tri-, or tetrasubstituted. The starting compounds 1 are easily prepared by reaction of the corresponding lithium enolates of alpha-chloroamides with aldehydes or ketones at - 78 degrees C. The influence of the reaction conditions and the structure of the starting compounds on the stereoselectivity of the beta-elimination reaction is also discussed. PMID- 11495434 TI - Oligosaccharides display both rigidity and high flexibility in water as determined by 13C NMR relaxation and 1H,1H NOE spectroscopy: evidence of anti-phi and anti-psi torsions in the same glycosidic linkage. AB - The trisaccharide beta-D-Glcp-(1-->2)-beta-D-Glcp-(1-->3)-alpha-D-Glcp-OMe has been investigated by molecular dynamics (MD) simulations and NMR experiments in water. 13C spin-lattice (T1) and spin-spin (T2) relaxation times, together with 1H,13C NOE data were measured at two magnetic field strengths (9.4 and 14.1 T) in a 277 K D2O solution. Relaxation data interpreted by means of the model-free formalism revealed a rigid (S2 approximately 0.9) oligosaccharide tumbling in solution. 1H,1H Cross-relaxation rates were determined at 600 MHz by 1D DPFGSE NOESY and T-ROESY experiments, which provided high quality data and subsequently proton-proton distances within the trisaccharide. The presence of anti conformers at both torsions of a glycosidic linkage is demonstrated for the first time. MD simulations were carried out to facilitate analysis of the NOE data. In total, 15 simulations-starting from five different conformational states--were performed, with production runs of up to 10 ns, resulting in 83 ns of oligosaccharide dynamics in water. anti Conformers were populated to different degrees in the simulations, especially at the phi2 torsion angle. By combining the results from the NOE experiments and the MD simulations, the anti conformers at the (1-->2) linkage were quantified as 7% anti-phi2 and 2% anti-psi2, revealing a highly flexible trisaccharide in which large conformational changes occur. From the MD simulations, interresidue hydrogen bonding, from HO2" to O2 or O3, was significantly populated (approximately 40%) in both of the anti conformational states. The contentious issue over rigidity versus flexibility in oligosaccharides has thus been thoroughly examined, showing that the dynamics should be taken into account for a relevant description of the molecular system. PMID- 11495435 TI - Homoleptic lanthanide amides as homogeneous catalysts for alkyne hydroamination and the Tishchenko reaction. AB - The homoleptic bis(trimethylsilyl)amides of Group 3 metals and lanthanides of the general type [Ln[N(SiMe3)2]3] (1) (Ln=Y, lanthanide) represent a new class of Tishchenko precatalysts and, to a limited extent, precatalysts for the hydroamination/cyclization of aminoalkynes. It is shown that 1 is the most active catalyst for the Tishchenko reaction. This contribution presents investigations on the scope of the reaction, substrate selectivity, lanthanide-ion size-effect, and kinetic/mechanistic aspects of the Tishchenko reaction catalyzed by 1. The turnover frequency is increased by the use of large-center metals and electron withdrawing substrates. The reaction rate is second order with respect to the substrate. While donor atoms, such as nitrogen, oxygen, or sulfur, on the substrate decrease the turnover frequency, 1 shows a tolerance for a large number of functional groups. For the hydroamination/cyclization of aminoalkynes, 1 is less active than the well-known metallocene catalysts. On the other hand, 1 is much more readily accessible (one-step synthesis or commercially available), than the metallocenes and might therefore be an attractive alternative catalyst. PMID- 11495436 TI - Chiral diphosphites derived from D-glucose: new ligands for the asymmetric catalytic hydroformylation of vinyl arenes. AB - A series of novel diphosphite ligands derived from readily available D-(+) glucose has been synthesized. These ligands have been applied to the Rh-catalyzed hydroformylation of vinyl arenes. Both excellent enantioselectivities (up to 91%) and regioselectivities (up to 98.8%) were achieved under mild conditions. The advantage of these ligands is that their modular natures allow facile, systematic variation in the configurations at the stereocenters [C(3), C(5)] at the ligand bridge and in the biphenyl substituents, enabling their effects on the stereoselectivity to be studied. Results show that the absolute configuration of the product is governed by the configuration at the stereogenic center C(3), while the level of the enantioselectivity is influenced by a cooperative effect between stereocenters C(3) and C(5). Replacement of the tert-butyl substituent by methoxy substituents at the para positions of the biphenyl moieties improved the enantioselectivities. We have characterized the rhodium complexes formed under CO/H2 by NMR techniques and in situ IR spectroscopy and have observed that there is a relationship between the structure of the [HRh(CO)2(PP)] species and their enantiodiscriminating performance in hydroformylation. Enantioselectivities were highest with ligands with a strong bis-equatorial coordination preference, while an equilibrium of species with bis-equatorial and equatorial-axial coordination modes considerably reduced the ee's. PMID- 11495437 TI - Synthesis and characterization of linear, hyperbranched, and dendrimer-like polymers constituted of the same repeating unit. AB - The synthesis of a linear polymer that includes both P=N and P=S double bonds, and P-O and P-C single bonds is reported by using two different paths that involve deprotection reactions and the Staudinger reaction. The preparation of hyperbranched polymers made up of OC6H4P(Ph)2=N-P=S repeating units is also described. Five generations of dendrimers originating from the same building blocks were prepared. The characterisation of all these phosphorus-based macromolecular architectures (solution behaviour, size exclusion chromatography, intrinsic viscosity, thermal behaviour) revealed marked differences in their respective behaviour. PMID- 11495438 TI - Phosphabenzenes as monodentate pi-acceptor ligands for rhodium-catalyzed hydroformylation. AB - A new class of phosphinine/rhodium catalysts for the hydroformylation of terminal and internal alkenes is presented in this study. A series of phosphabenzenes 1-14 has been prepared by condensation of phosphane or tris(trimethylsilyl)phosphane with the corresponding pyrylium salt. Trans-[(phosphabenzene)2RhCl(CO)] complexes 21-25 have been prepared and studied spectroscopically and by X-ray crystal structure analysis. The hydroformylation of oct-1-ene has been used to identify optimal catalyst preformation and reaction conditions. Hydroformylation studies with 15 monophosphabenzenes have been performed. The catalytic performance is dominated by steric influences, with the phosphabenzene 8/rhodium system being the most active catalyst. Turnover frequencies of up to 45370 h(-1) for the hydroformylation of oct-1-ene have been determined. In further studies, hydroformylation activity toward more highly substituted alkenes was investigated and compared with the standard industrial triphenylphosphane/rhodium catalyst. The reactivity differences between the phosphabenzene and the triphenylphosphane catalyst increase on going to the more highly substituted alkenes. Even tetrasubstituted alkenes reacted with the phosphabenzene catalyst, whereas the triphenylphosphane system failed to give any product. In situ pressure NMR experiments have been performed to identify the resting state of the catalyst. A monophosphabenzene complex [(phosphinine 8)Ir(CO)3H] could be detected as the predominant catalyst resting state. PMID- 11495439 TI - Mechanistic aspects of the reaction between Br2 and chalcogenone donors (LE; E=S, Se): competitive formation of 10-E-3, T-shaped 1:1 molecular adducts, charge transfer adducts, and [ (LE)2]2+ dications. AB - The synthesis and spectroscopic characterisation of the products obtained by treatment of N,N'-dimethylimidazolidine-2-thione (1), N,N'-dimethylimidazolidine 2-selone (2), N,N'-dimethylbenzoimidazole-2-thione (3) and N,N' dimethylbenzoimidazole-2-selone (4) with Br2 in MeCN are reported, together with the crystal structures of the 10-E-3, T-shaped adducts 2 . Br2 (12), 3 . Br2 (13) and 4 . Br2 (14). A conductometric and spectrophotometric investigation into the reaction between 1-4 and Br2, carried out in MeCN, allows the equilibria involved in the formation of the isolated 10-E-3 (E = S, Se) hypervalent compounds to be hypothesised. In order to understand the reasons why S and Se donors can give different product types on treatment with Br2 and I2, DFT calculations have been carried out on 1-8, 19 and 20, and on their corresponding hypothetical [LEX]+ cations (L = organic framework; E = S, Se; X = Br, I), which are considered to be key intermediates in the formation of the different products. The results obtained in terms of NBO charge distribution on [LEX]+ species explain the different behaviour of 1-8, 19 and 20 in their reactions with Br2 and I2 fairly well. X-ray diffraction studies show 12-14 to have a T-shaped (10-E-3; E = S, Se) hypervalent chalcogen nature. They contain an almost linear Br-E-Br (E = S, Se) system roughly perpendicular to the average plane of the organic molecules. In 12, the Se atom of each adduct molecule has a short interaction with the Br(1) atom of an adjacent unit, such that the Se atom displays a roughly square planar coordination. The Se-Br distances are asymmetric [2.529(1) vs. 2.608(1) A], the shorter distance being that with the Br(1) atom involved in the short intermolecular contact. In contrast, in the molecular adducts 13 and 14, which lie on a two-fold crystallographic axis, the Br-E-Br system is symmetric and no short intermolecular interactions involving chalcogen and bromine atoms are observed. The adducts are arranged in parallel planes; this gives rise to a graphite-like stacking. The new crystalline modification of 10, obtained from acetonitrile solution, confirms the importance of short intermolecular contacts in determining the asymmetry of Br-E-Br (E = S, Se) and I-Se-I groups in hypervalent 10-E-3 compounds. The analogies in the conductometric and spectrophotometric titrations of 1 and 2-4 with Br2, together with the similarity of the vibrational spectra of 11-14, also imply a T-shaped nature for 11. The vibrational properties of the Br-E-Br (E = S, Se) systems resemble those of the Br3- and IBr2- anions: the Raman spectrum of a symmetric Br-E-Br group shows only one peak near 160 cm(-1), as found for symmetric Br3- and IBr2- anions, while asymmetric Br-E-Br groups also show an antisymmetric Br-E-Br mode at around 190 cm(-1), as observed for asymmetric Br3- and IBr2- ions. Therefore, simple IR and Raman measurements provide a useful tool for distinguishing between symmetric and asymmetric Br-E-Br groups, and hence allow predictions about the crystal packing of these hypervalent chalcogen compounds to be made when crystals of good quality are not available. PMID- 11495440 TI - Modified windmill porphyrin arrays: coupled light-harvesting and charge separattion, conformational relaxation in the S1 state, and S2-S2 energy transfer. AB - The architecture of windmill hexameric zinc(II) -porphyrin array 1 is attractive as a light-harvesting functional unit in view of its three-dimensionally extended geometry that is favorable for a large cross-section of incident light as well as for a suitable energy gradient from the peripheral porphyrins to the meso-meso linked diporphyrin core. Three core-modified windmill porphyrin arrays 2-4 were prepared for the purpose of enhancing the intramolecular energy-transfer rate and coupling these arrays with a charge-separation functional unit. Bisphenylethynylation at the meso and meso' positions of the diporphyrin core indeed resulted in a remarkable enhancement in the intramolecular S1-S1 energy transfer in 2 with tau=2 approximately 3 ps, as revealed by femtosecond time resolved transient absorption spectroscopy. The fluorescence lifetime of the S2 state of the peripheral porphyrin energy donor determined by the fluorescence up conversion method was 68 fs, and thus considerably shorter than that of the reference monomer (150 fs), suggesting the presence of the intramolecular energy transfer channel in the S2 state manifold. Such a rapid energy transfer can be understood in terms of large Coulombic interactions associated with the strong Soret transitions of the donor and acceptor. Picosecond time-resolved fluorescence spectra and transient absorption spectra revealed conformational relaxation of the S1 state of the diporphyrin core with tau = 25 ps. Upon photoexcitation of models 3 and 4, which bear a naphthalenetetracarboxylic diimide or a meso-nitrated free-base porphyrin attached to the modified diporphyrin core as an electron acceptor, a series of photochemical processes proceeded, such as the collection of the excitation energy at the diporphyrin core, the electron transfer from the S1 state of the diporphyrin to the electron acceptor, and the electron transfer from the peripheral porphyrins to the diporphyrin cation radical, which are coupled to provide a fully charge-separated state such as that in the natural photosynthetic reaction center. The overall quantum yield for the full charge separation is better in 4 than in 3 owing to the slower charge recombination associated with smaller reorganization energy of the porphyrin acceptor. PMID- 11495441 TI - Tellurium-bridged manganese carbonyl clusters: synthesis and structural transformations of [Te4Mn3(CO10]-, [Te2Mn3(CO)9]2-, [Te2Mn3(CO)9]-, and [Te2Mn4(CO)12]2-. AB - The reactions of appropriate ratios of K2TeO3 and [Mn2(CO)10)] in superheated methanol solutions lead to a series of novel cluster anions [Te4Mn3(CO)10] (1), [Te2Mn3(CO)9]2- (2), [Te2Mn3(CO)9]- (3), and [Te2Mn4(CO)12]2- (4). When cluster 1 is treated with [Mn2(CO)10]/KOH in methanol, paramagnetic cluster 2 is formed in moderate yield. Cluster 2 is oxidized by [Cu(MeCN)4]BF4 to give the closo-cluster [Te2Mn3(CO)9]- (3), while treatment of 2 with [Mn2(CO)10]/KOH affords the closo cluster 4. IR spectroscopy showed that cluster 1 reacted with [Mn2(CO)10] to give cluster 4 via cluster 2. Clusters 1-4 were structurally characterized by spectroscopic methods or/and X-ray analyses. The core structure of 1 can be described as two [Mn(CO)3] groups doubly bridged by two Te2 fragments in a mu2 eta2 fashion. Both [Mn(CO)3] groups are further coordinated to one [Mn(CO)4] moiety. Cluster 2 is a 49 e- species with a square-pyramidal core geometry. While cluster 3 displays a trigonal-bipyramidal metal core, cluster 4 possesses an octahedral core geometry. PMID- 11495442 TI - Di- and tetranuclear palladium complexes incorporating phospha- and diphosphaferrocenes as ligands. AB - Monophosphaferrocenes and 4 react with [Pd(COD)Cl2] (COD = cyclooctadiene) to afford cis- [Pd(1 or 4)2Cl2] complexes that slowly decompose in solution to give dimeric complexes 3 and 6 of general formula [[Pd(1 or 4)Cl]2]. In these dimers, which incorporate a Pd-Pd bond, phosphaferrocenes act as four electron donors through the phosphorus-atom lone pair (mu2-bonded) and through one orbital of appropriate symmetry at iron. These dimers can also be more conventionally prepared from the reaction of cis- [Pd(1 or 4) Cl2] complexes with [Pd(dba)2] (dba = dibenzylidene acetone). The reaction of octaethyldiphosphaferrocene (7) with [Pd(COD)Cl2] yields a dinuclear complex [Pd2(7)2Cl4] (8) in which the two ligands 7 are coordinated in a trans fashion through the phosphorus-atom lone pairs. Decomposition of 8 in solution yields a dimeric dicationic complex of general formula [[Pd2(7)2Cl]2]2+[FeCl4]2- (9a) incorporating four palladium atoms. In each ligand. one phospholyl ring behaves as a two-electron donor through the phosphorus-atom lone pair whereas the second binds two palladium centers in a mu2-fashion. A plausible mechanism that explains the formation of dimers 3, 6, and 9a involves the preliminary oxidation of the mono- or diphosphaferrocene ligand. Parallel experiments aimed at confirming this hypothesis have shown that complex 9a can be synthesized from the reaction of FeCl2 with complex 8. Also presented is another synthetic approach to the synthesis of the tetranuclear complex 9b (counterion is GaCl4-) from the reaction of the palladium(0) complex [Pd(7)2] (10) with [Pd(COD)Cl2] the presence of GaCl3 as chloride abstractor. PMID- 11495443 TI - The stability of gold iodides in the gas phase and the solid state. AB - The stability of gold iodides in the oxidation state +I and +III is investigated at the ab initio and density functional level using relativistic and nonrelativistic energy-adjusted pseudopotentials for gold and iodine. The calculations reveal that relativistic effects stabilize the higher oxidation state of gold as expected, that is Au2I6 is thermodynamically stable at the relativistic level, whilst at the nonrelativistic level the complex of two iodine molecules weakly bound to both gold atoms in Au2I2 is energetically preferred. The rather low stability of AuI3 with respect to dissociation into AuI and I2 will make it difficult to isolate this species in the solid state as (possibly) Au2I6 or detect it by matrix-isolation techniques. The monomer AuI3 is Jahn Teller distorted from the ideal trigonal planar (D3h) form, but adopts a Y-shaped structure (in contrast to AuF3 and AuCl3), and in the nonrelativistic case can be described as I2 weakly bound to AuI. Relativistic effects turn AuI3 from a static Jahn-Teller system to a dynamic one. For the yet undetected gas-phase species AuI accurate coupled-cluster calculations for the potential energy curve are used to predict vibrational-rotational constants. Solid-state density functional calculations are performed for AuI and Au2I6 in order to predict cohesive energies. PMID- 11495444 TI - Cataract: the main cause of blindness in leprosy. PMID- 11495445 TI - The threshold for protective sensation that prevents neuropathic ulceration on the plantar aspect of the foot: a study of leprosy patients in a rural community in India. AB - The protective sensation threshold is an important concept in the prevention of plantar ulceration in leprosy patients. Previous studies have suggested that skin with sensory nerve damage on the plantar aspect of the foot which can still detect the 5.07 Semmes-Weinstein monofilament (approximately 10 g) is highly unlikely to develop ulceration. While the threshold is thought to be less than the 610 filament (approximately 75 g), no work just testing adjacent to current ulcers has been undertaken to assess this more accurately. This is important, as it has been shown that a significant proportion of healthy individuals who wear sandals or go barefoot in India may fail to detect this 5.07 filament in at least some areas of the sole, especially in older age groups, and in certain cases the 5.46 filament (approximately 30 g) is the lightest detected. In an attempt to address this problem, a cross-sectional study on 26 current plantar ulcers in male adults with stable neuropathy due to leprosy was carried out in the rural town of Salur, India. It was confirmed that the ability to detect the 5.07 filament (approximately 10 g) did prevent the development of ulceration while in contrast the ability to detect the 5.46 filament (approximately 30 g) did not. This suggests that the threshold for protective sensation lies between these two filaments. An approach is suggested which may help to differentiate feet genuinely at risk of ulceration from those merely unable to detect the 5.07 filament on account of thickened skin callus or advancing age. PMID- 11495446 TI - Can people affected by leprosy at risk of developing plantar ulcers be identified? A field study from central Ethiopia. AB - In the ALERT leprosy control programme, 75 people affected by leprosy, in three different geographical areas, were investigated. Each person was documented as having anaesthesia to the 10 g monofilament. The study sought to determine why some people developed ulcers whilst others did not. According to the records, 43 had an ulcer during the last 5 years but 32 had never had an ulcer. In order to examine protective sensation on the sole of the foot, various sensory modalities were tested and the functional anatomy of the foot was examined. The results showed, as may be expected, that it is not possible to define a specific threshold for protective sensation that could be applied to all cases. Some people with only slightly diminished sensation developed ulcers, while many others with almost complete anaesthesia remained ulcer-free. In these rural communities, being a farmer reduced the risk of developing an ulcer, but no other demographic features were significant. Graded monofilaments were found to be the most appropriate test, with loss of sensation at any of five points tested being a 'positive' result. The 10 g filament was the most sensitive, but only 43% of feet identified by this test actually developed an ulcer. As people with partial loss of sensation were excluded from this study, this figure may be lower under programme conditions. The 50 g and 100 g filaments decrease the number of feet identified as at risk, but increase the percentage which actually develop an ulcer, to 46% and 49%, respectively. An appropriate test for selecting those for special programmes which may have a limited capacity, for example the provision of subsidized footwear or involvement in self-care groups, would be a 100 g filament, which would detect 86% of those feet likely to develop an ulcer, while reducing the number of those selected who are not at great risk. Vibrometry was found to be no better than graded filaments and an examination of functional anatomy did not help in identifying those at risk. PMID- 11495447 TI - Dynamics of impairment during and after treatment: the AMFES cohort. AB - This study investigates the dynamics of impairment during and after multidrug therapy treatment for the patient cohort of the prospective ALERT MDT Field Evaluation Study (AMFES). The impairment status was compared at intake, at release from treatment (rft), and at the time of the latest survey between 24 and 48 months after release from treatment (follow-up). The eye-hand-foot impairment score (EHF score), which is the sum of the WHO impairment grades of the eyes, hands, and feet, was used as tool for comparison. In all, 433 out of the 592 patients (224 PB and 209 MB) completed treatment in time and were assessed at release from treatment. The risk of getting impaired was 4% for the 113 PB and 21% for the 91 MB patients who were initially free from impairment. Out of the 111 initially impaired PB patients, 41% recovered or improved and 13% worsened in EHF score. For the 118 initially impaired MB patients, these figures were: recovery or improvement 43% and worsening 13%. Three hundred and twenty-three out of the 433 patients (158 PB and 165 MB) had a follow-up examination in between the next 24-48 months after rft. The risks of impairment at follow-up were 6% for the 79 PB and 18% for the 77 MB patients without impairment at rft. Out of the 79 PB patients with impairment at rft, 35% recovered or improved and 28% worsened. For the 88 impaired MB patients, these figures were: recovery or improvement 26% and worsening 27%. Patients showed a tendency to compensate EHF score improvement before rft by worsening after rft and vice versa. The first main conclusion is that the impairment status at intake was by far the most important determinant for future impairment. The second one is that the dynamics of impairment were less favourable after rft than before. Little is known about the long-term fate of leprosy patients with irreversible nerve damage and the associated risk of developing severe secondary impairment. Especially in this era of the leprosy elimination goal, we should give this accumulating patient group due attention in research and health policy agendas. PMID- 11495448 TI - Fine needle aspiration cytology (FNAC) of nerves in leprosy. AB - Leprosy is primarily a disease of the peripheral nerves and a technique that is simpler than nerve biopsy is required to evaluate nerve involvement, especially in pure neuritic (PN) leprosy. This study was designed to evaluate the role of FNAC of the nerve in the diagnosis and classification of leprosy. A prospective study was carried out on 25 patients with clinically active leprosy and at least one thickened peripheral sensory nerve. Nerve aspirates were evaluated by May Grunwald-Giemsa and Fite's staining. Lepromin test, slit skin smears (SSS), skin biopsies (except PN cases) and nerve biopsies were performed and compared with FNAC. FNAC of nerve from 23 cases (92%) yielded diagnostic aspirates. Acid fast bacilli were observed in six cases by FNAC. FNAC and nerve pathology were equally comparable with the other parameters evaluated. Based on the results, cytological criteria were developed for interpreting nerve aspirates and the cases were classified as paucibacillary (18), BB (2), BL (2), LL (1) and non-diagnostic (2). All PN cases showed diagnostic paucibacillary type cytology. FNAC of the nerve yields diagnostic aspirates in leprosy comparable with nerve pathology and the proposed cytological criteria may be useful in classification of nerve aspirates. PMID- 11495449 TI - Response of Mycobacterium habana vaccine in patients with lepromatous leprosy and their household contacts. A pilot clinical study. AB - Single dose vaccination was carried out with Mycobacterium habana vaccine, 31 lepromatous leprosy cases receiving 1.5 mg (1.5 mg = 6.27 x 10(8) bacilli) and 36 household contacts randomly receiving 1.5, 2.0, 2.5 mg vaccine intradermally. Duration of study was 18 weeks. Vaccination induced lepromin conversion in 100% of lepromatous leprosy cases and lepromin negative household contacts and augmentation of lepromin reactivity in 100% of lepromin positive household contacts, which was stable for the 15 weeks duration of follow-up. The maximum augmentation in lepromin reactivity was obtained with 1.5 mg of vaccine, which is probably the supramaximal dose. Overall, post-vaccination, those without prior BCG vaccination scars showed higher mean values of lepromin augmentation. Local vaccination site changes included induration, ulceration, itching, pain and uncomplicated regional lymphadenopathy, all of which remitted spontaneously by 15 weeks. Systemic side-effects noted were pyrexia, ENL and jaundice, and were seen with no greater frequency than that reported in other vaccine trials. Overall, systemic side-effects were easily controlled and were not accompanied by clinically detectable nerve or ocular damage. The safety profile investigations revealed an increase in the mean values of Hb%, RBC count and PCV in household contacts and of PCV in lepromatous patients, post-vaccination. Alterations in the liver function tests were also observed in patients of lepromatous leprosy. Thus, M. habana vaccine appears to be useful in stimulating specific CMI against M. leprae as evidenced by increased lepromin reactivity. PMID- 11495450 TI - Treatment outcome and impact of leprosy elimination campaign in Sokoto and Zamfara states, Nigeria. AB - A Leprosy Elimination Campaign (LEC) was implemented in 37 districts of Sokoto and Zamfara states, Nigeria from 13 August to 30 November 1998. The campaign utilized intensive community mobilization and training of local health personnel to detect hidden leprosy cases. During 8 weeks of case finding, 160,127 persons were screened; 353 new cases of leprosy were detected and placed on MDT; 236 (67%) of new cases detected were classified as MB, 64 cases (18%) suffered visible deformities and 24 patients (6.8%) were children. Follow-up in December 1999 of patients placed on MDT revealed 97% PB and 96% MB cure rates, respectively. Detection of cases in communities led some community leaders to ask for repeat surveys in their communities. Repeat surveys continue to yield new cases. The authors recommend that LECs be maintained for 3 years to accelerate leprosy elimination in the region. The cost effectiveness and impact of LEC in Sokoto-Zamfara are discussed. PMID- 11495451 TI - Defaulting patterns in a provincial leprosy control programme in Northern Mozambique. AB - Cohort-based multidrug therapy (MDT) completion rates are used to assess adherence to MDT. However this measure gives no information about when during the treatment period defaulting occurs. Two districts in Cabo Delgado province in Northern Mozambique were selected for evaluation of multibacillary patient defaulter data between 1993 and 1997 to examine when patients default during the treatment penod. In all, 548 (59.2%) of 926 MB patients completed treatment and 378 (40.8%) defaulted between 1993 to 1997. The percentage of defaulters fell steadily from 59.8% in 1993 to 23.2% in 1997. Of the 378 defaulters 57.7% defaulted treatment within 6 months and 83.1% within 1 year of starting treatment. It was observed that patients tend to default early rather than late in the treatment period and that this pattern is maintained over time despite a fall in defaulter rates. Patients established early into a treatment routine were more likely to complete treatment. A comprehensive effort to improve and maintain leprosy control services will probably influence adherence more than any single, specific strategy. Shortening MDT treatment from 2 years to 1 year is unlikely to affect the defaulter rate. PMID- 11495452 TI - Should nutritional status evaluation be included in the initial needs assessment of leprosy patients with disability prior to socio-economic. AB - Because of the large numbers of leprosy patients with disability and the limited resources available, it is important that socio-economic rehabilitation (SER) is targeted towards those who are most in need. Towards this purpose, current assessments of leprosy patients prior to initiating SER include the evaluation of income, assets and household possessions. Conspicuously absent is the nutritional assessment of the patient. In the absence of weight loss associated with illness, population studies indicate that undernutrition reflects poor socio-economic conditions. In this study of 151 cured leprosy patients with disability, 57% of the patients were found to be undernourished using body mass index (kg/m2) derived from body weight and height, and 10% of the patients were severely undernourished (grade III). Undernutrition in the patients was poorly though significantly correlated with personal income (r = 0.18, P < 0.05). Total household income, reported amount of money spent on food and estimated cereal intakes were not correlated with the BMI of the patient, possibly due to reporting bias and other methodological issues. We propose the inclusion of nutritional status evaluation by anthropometry as part of the initial screening of leprosy patients prior to instituting SER. We believe that this simple and objective evaluation can add to the assessment of 'threat' of economic deprivation or actual economic 'dislocation', and thus help in the prioritization of leprosy patients for SER. PMID- 11495453 TI - The biology of HIV infection. AB - This article reviews the cell and molecular biology of human immunodeficiency virus (HIV), emphasizing the features that lead to opportunistic infection by organisms such as mycobacteria. Mycobacteria, especially M. avium complex and M. tuberculosis infections, are closely associated with HIV disease. HIV is a very small retrovirus and its high mutation rate leads to extremely variable viral populations, both within and between individuals. It is coated with glycoprotein 120 (gp120), which it uses to bind to and infect a range of CD4+ leukocytes, depending on the co-receptor specificity. T cell-tropic HIV strains tend to use the CXCR-4 chemokine receptor, while macrophage-tropic strains tend to use the CCR-5 chemokine receptor. Immunosuppression is induced in a number of ways. As well as frank depletion of virus-infected T cells, antigen-specific T cell clones can be selectively deleted by mechanisms such as defective antigen presentation by HIV-infected macrophages (activation-induced cell death). Changes in cytokine production in HIV infection are also proposed. All this leads to falling T cell counts, B cell dysregulation and macrophage dysfunction. Opportunistic infections exploit this immunosuppressed environment. Certain infections are prevalent, reflecting factors such as environmental exposure to pathogens, poor mucosal defences and subcellular interactions between HIV and, e.g. viral or mycobacterial infections. Opportunistic infection exacerbates immune destruction by HIV, producing a vicious cycle that is ultimately fatal. PMID- 11495454 TI - Comparative value of active and passive surveillance over time in treated leprosy patients, in the prevention of further disability. PMID- 11495455 TI - Analysis of genetic relationships between various populations of domestic and jungle fowl using microsatellite markers. AB - The genomes of domestic and jungle fowl populations maintained in Ukraine and Germany were screened using microsatellites as molecular markers. Genetic variation and genetic distances between strains of different origins and performance potentials were determined. In total, 224 individuals of 20 populations were genotyped for 14 microsatellite markers covering 11 linkage groups. Of the 14 microsatellite loci, the number of alleles ranged between 2 and 21 per locus, the mean number of alleles being 11.2 per locus. By using Nei's standard distance and the Neighbor-Joining method, a phylogenetic tree was reconstructed; its topology reflected general patterns of relatedness and genetic differentiation among the chicken populations studied. Three major phylogenetic tree groupings were found. The red jungle fowl (Gallus gallus) formed a separate branch and demonstrated a specific allele distribution when compared with domestic fowl breeds analyzed. The second branch comprised commercial layer lines and chicken breeds that were subject to intense selection in the past or had common ancestral breeds with commercial strains. The third group encompassed the German native breed populations. The information about population and breed genetic relationships estimated by microsatellite analysis may be useful as an initial guide in defining objectives for designing future investigations of genetic variation and developing conservation strategies. PMID- 11495456 TI - Resistance to Marek's disease virus in White Leghorn chickens: effects of avian leukosis virus infection genotype, reciprocal mating, and major histocompatibility complex. AB - Genetic improvement for resistance to Marek's Disease (MD) in chickens continues to be of interest to the poultry industry. The aims of this study were to identify effects of the MHC on the molecular level and of avian leukosis virus (ALV) resistance status on MD mortality in two noninbred White Leghorn chicken lines that differ in B blood group type. Previously, within each of the chicken lines, sublines had been selected for resistance or susceptibility to ALV infection with Subgroups A and B. In this study, F2 offspring, obtained by crossing the two ALV-resistant or the two ALV-susceptible sublines, were tested for MD mortality after contact exposure at 1 d of age. Reciprocal matings were made in the grandparental generation. The MD mortality percentages, in an observation period of 17 wk, of F2 offspring from two hatches were 82.63 and 92.35%, respectively. Survival analysis (Cox model) was applied to assess the risk of dying from MD. No differences in MD mortality risk profiles were found between ALV-resistant and ALV-susceptible F2 offspring. Within ALV-susceptible F2 offspring, however, a reciprocal mating effect was observed in both hatches. The MHC Class I, II, and IV restriction fragment length polymorphism (RFLP) analyses were carried out on birds of the first hatch. Although two of 11 MHC class IV RFLP bands displayed a significant effect, in general, a strong association of MHC and MD mortality was not detectable. PMID- 11495457 TI - Effect of route of inoculation on humoral immune response of White Leghorn chickens selected for high or low antibody response to sheep red blood cells. AB - Effects of route of SRBC inoculation and antigen dosage on primary and secondary antibody response of White Leghorn lines selected for high (HA) or low (LA) 5-d antibody response to a single i.v. inoculation with 0.1 mL of a 0.25% suspension of SRBC were studied in two trials. In the first trial, chicks from parents of generation S24 of each line were randomly assigned to one of four treatments. At 35 d of age, they were inoculated into the brachial vein with 0.1 mL of 0.25% suspension of SRBC or into the breast muscle with 0.1 mL of 0.25, 2.50, or 25.00% SRBC. Plasma SRBC antibody was measured 3, 6, 10, and 20 d later. In the second trial, chicks from parents of generation S25 of each line were randomly assigned to treatment groups. At 28 d of age they were inoculated with 0.1 mL of 0.25% SRBC into the brachial vein, 0.1 mL of 25.00% SRBC into the thigh (T-L) or breast muscle (B-L), or 0.5 mL of 25.00% SRBC into the thigh (T-H) or breast muscle (B H). Twenty-one days later, chicks (except five per group) were given a booster inoculation of 0.1 mL of 25.00% SRBC into the thigh muscle. Six and 10 d after each inoculation, plasma SRBC antibody, IgG, and IgM titers were measured. The SRBC antibody titers after primary i.v. inoculation with SRBC were always higher for HA than LA chicks. When inoculations were i.m., differences between lines varied with dosage. Low dosages inoculated into the breast failed to induce line differences consistently, whereas at higher dosages, titers were greater for HA than LA chicks regardless of inoculation site. For Line LA, inoculation into the thigh elicited higher titers than inoculations into the breast. Antibody titers to the booster inoculation of SRBC were similar for the lines. PMID- 11495458 TI - Effect of genetic selection for group productivity and longevity on immunological and hematological parameters of chickens. AB - A line of White Leghorn chickens was selected for high group productivity and longevity resulting in improved survival and feather score as well as reduced cannibalism and flightiness. Improvements in survival might have also been due to improved immunity. The present study was designed to test the hypothesis that selection for high (HGPS) and low (LGPS) group productivity and survivability also altered immune and hematological parameters. The LGPS line was an intense reverse selected line of the HGPS line at the eighth generation of development. Hens were randomly assigned to individual cages at 17 wk of age. Blood samples were collected from the hens at 21 wk of age. Subsets of T lymphocytes (CD4+, CD8+, and gammadelta cells) were measured using flow cytometry. Concentrations of plasma IgG were quantified with western blot analysis and immunoprecipitation assay. Hematological parameters were collected from blood smears. The HGPS hens had significantly higher percentages of blood lymphocytes and CD4+:CD8+ ratios of circulating T cells (P < 0.01) and tended to have more, but not significantly, gammadelta T cells (P = 0.07) than the LGPS hens. In contrast, the LGPS hens exhibited eosinophilia and heterophilia and greater heterophil:lymphocyte ratios (P < 0.01). The concentrations of plasma IgG were also significantly higher in the LGPS hens (P < 0.01). These results suggest that genetic selection for group productivity and longevity also alters the immunological and hematological systems of hens. The line difference in regulation of T cells, leukocytes, and production of IgG may suggest that different genes or modes of gene action are involved. PMID- 11495459 TI - Genetic variation in pure lines and crosses of large-bodied turkey lines. 1. Body weight, walking ability, and body measurements of live birds. AB - An experimental line of turkeys selected for increased BW (F) was reciprocally crossed with sire lines (designated A and B) from each of two major commercial breeders in order to study the inheritance of growth traits. All genetic groups were grown intermingled in confinement with the sexes reared in different houses. Traits measured included BW at 8, 16, and 20 wk of age; shank length, width, and depth; breast width; and walking ability scores at 16 wk of age. The F line had a different growth pattern than the two commercial sire lines. The F line had higher (males) or similar (females) BW in comparison to Line A at 8 wk of age, but by 16 wk of age, Line A was heavier than Line F in both sexes. The difference in BW between the F and A lines increased from 16 to 20 wk of age. Lines F and B did not differ in BW at 8 wk of age, but at 16 and 20 wk of age, Line B birds were heavier than those of F line. Reciprocal effects, a measure of sex linkage and maternal effects, were noted only for breast width of females in crosses of Lines A and F and for shank width and depth of males and 8-wk BW and shank depth of females in crosses of Lines B and F. Heterosis was an important source of variation in BW for males from both crosses. The percentage heterosis at the various ages ranged from 3.1 to 7.5. For females, heterosis (range = 2.6 to 4.9%) was only significant at younger ages (8 wk for the crosses of the A and F lines and 8 and 16 wk for the crosses of the B and F lines). No heterosis for breast width was observed in either cross. Heterosis in walking ability scores was significant only for males from the crosses of Lines B and F. The presence of heterosis was inconsistent for shank measurements. PMID- 11495460 TI - Genetic variation in pure lines and crosses of large-bodied turkey lines. 2. Carcass traits and body shape. AB - An experimental line (F) of turkeys was reciprocally crossed with sire lines (designated A and B) from each of two commercial breeders in order to study the inheritance of carcass traits and body shape. The birds were weighed and killed at 17 wk of age, and various measurements of muscling, leg bones, and body shape were made. Additive genetic variation, as indicated by line differences, was an important source of variation in most traits. The only traits that did not exhibit a line difference in any comparison were weight of the drumstick muscles, tarsometatarsal width, keel length, and Body Depth 2 (body depth at a point 2.54 cm anterior to the posterior end of the keel). Heterosis of live BW was greater in males than in females. Heterosis in the weight of the pectoralis major and p. minor muscles were similar to those of live BW, but heterosis in the weight of the leg muscles were higher than that of live BW. The average heterosis for bone measurements (length of the femur, tarsometatarsal, and tibiotarsal bones and tarsometatarsal width) was very low and ranged from -0.3 to 1.4%. For measurements of body shape [keel length, Body Depth 1 (body depth measured at the cranial process of the keel), Body Depth 2, ratio of Body Depth 1 to Body Depth 2, length, width, and height of the body cavity, and body cavity volume index), heterosis was low and the average ranged from -2.7 to 2.6%. Based on an analysis of both sexes combined, the commercial sire lines differed in BW, weights of the p. minor and drumstick muscles, weights of the tarsometatarsal, femur, and tibiotarsal bones, lengths of the tarsometatarsal and femur bones, and various measurements of body shape (Body Depth 1, body depth ratio, body cavity length and height, and body cavity volume index). Relative to the commercial sire lines, the F line was smaller, had less breast and leg muscling, and, in general, larger leg bones when sexes were combined. The carcass of the F line was deeper than that of the commercial sire lines when measured at the cranial process of the keel but not at 2.54 cm anterior to the posterior portion of the keel. Body cavity height and body cavity volume index were larger in the F line than in the commercial sire lines. PMID- 11495461 TI - Genetic line differences in survival and pathogen load in young layer chicks after Salmonella enterica serovar enteritidis exposure. AB - Early infection may result in long-term colonization of layers with Salmonella enterica sv. enteritidis (S. enteritidis, SE), resulting in shedding into table or hatching eggs. To evaluate genetic factors underlying early response to SE, genetic line differences in mortality and pathogen load at two sites (cecal lumen and spleen) were investigated. At day of hatch, chicks of four genetic lines were intra-esophageally inoculated with one of three doses of SE phage type 13a. There was a significant effect (P < 0.001) of genetic line on chick 6-d survival. The effect of genetic line was significant (P < 0.05) on survivors' SE burden in cecal content but not on SE burden per gram of spleen. The SE pathogen load of the spleen and the cecal content were not significantly correlated, indicating that independent host mechanisms are partly responsible for these two traits. Genetic line differences in chick survival and SE colonization of cecal content were demonstrated in young layer chicks. PMID- 11495462 TI - Identification of new major histocompatibility complex class II restriction fragment length polymorphisms in a closed experimental line of Beltsville Small White turkeys. AB - Beltsville Small White (BSW) turkeys have been utilized as an experimental model in the study of bacterial, parasitic, and fungal diseases. Given the critical role of MHC antigens in the initial steps of the immune response to specific pathogens, the MHC Class II of BSW turkeys was characterized. Southern blot analysis of PvuII-digested turkey DNA that was hybridized with a chicken Class II beta gene genomic clone revealed two restriction fragment length polymorphism profiles not previously identified in experimental or commercial breeder lines of turkeys. These fingerprint profiles differed in a single 6.0-kb band that was present in approximately 38% of the birds examined. The DNA fragments of 5.0, 4.1, 3.3, and 3.1 were present in both profiles. Furthermore, no mixed lymphocyte reaction was observed between individuals within the BSW turkey line. The present results indicate that BSW turkeys represent a unique source of genetic diversity for MHC Class II haplotypes. PMID- 11495463 TI - Effects of light intensity from photostimulation in four strains of commercial egg layers: 1. Ovarian morphology and carcass parameters. AB - We examined the effects of light intensity (LI) from photostimulation to 45 wk of age on egg production parameters and egg size characteristics and on ovarian and carcass morphology at sexual maturity and 45 wk of age in four layer strains. Floor housed pullets were raised in a light-tight facility from 1 d of age until housing in individually illuminated cages at 17 wk of age. Two white egg strains, ISA-White (ISA-W) and Shaver 2000 (S2000), and two brown egg strains, ISA-Brown (ISA-B) and Shaver 579 (S579), were used. Pullets were randomly assigned to a processing group that was killed at sexual maturity (first oviposition) (Group 1) or kept to 45 wk (Group 2). Birds were photostimulated at 18 wk of age using a LI of 1, 5, 50, or 500 lx (4 x 4 factorial design). One bird from Group 1 and one from Group 2 were caged together in individually lit cages (one brown and one white egg layer). Carcass and ovarian morphology data were examined as related to Strain, LI, or the interaction of Strain and LI. The time from photostimulation to sexual maturity did not differ due to LI, but was shorter for brown egg strains (ISA-B = 19.9 d, S579 = 20.2 d) than for white egg strains (ISA-W = 26.6 d, S2000 = 28.1 d). Body weight at sexual maturity differed among all strains, with the white egg strains having the lowest BW. Ovary weight was the greatest in ISA-W birds, in which 8.0 large yellow follicles (LYF) were present compared to 6.8 in S2000 birds. The LI affected ovary development, as birds with the 1 lx exposure had lower ovary weights and fewer LYF than did 50 lx birds, suggesting that the 1 lx LI did not result in an adequate photostimulatory cueing of sexual maturation. The threshold LI for a complete morphological response to photostimulation in this study was 5 lx. Strain differences in BW observed at sexual maturity continued to 45 wk of age. Light intensity affected 45 wk BW, with 5 lx LI birds weighing 7.2 and 8.7% more than the 50 and 500 lx birds, respectively. On an absolute basis, brown egg stains carried significantly more breast muscle at 45 wk of age than did white egg strains. The fatpad was heavier on a relative basis for brown egg layers than white egg layers. The 1 lx hens had lower 45-wk ovary weights than did the other three LI treatments. These data support the conclusion that with the development of highly specific genetic strains, it is increasingly important to match the environmental management practices to a particular hen's genotype. PMID- 11495464 TI - Effects of light intensity from photostimulation in four strains of commercial egg layers: 2. Egg production parameters. AB - The effects of light intensity (LI) from photostimulation to 45 wk of age on egg production parameters and egg size characteristics were examined in four layer strains. Floor housed pullets were raised in a light-tight facility from 1 d of age until housing in individually illuminated cages at 17 wk of age. At 17 wk of age, two white egg strains, ISA-White (ISA-W) and Shaver 2000 (S2000), and two brown egg strains, ISA-Brown (ISA-B) and Shaver 579 (S579), were assigned to a processing group [Group 1 was killed at sexual maturity (first oviposition); Group 2 was kept to 45 wk] and were photostimulated at 18 wk of age using a LI of 1, 5, 50, or 500 lx (4 x 4 factorial design). One bird from Group 1 and one bird Group 2 were caged together in individually lit cages (one brown and one white egg layer). Cages were equipped with hardware to monitor egg laying time. Data of individual egg weight and time of lay were kept on Group 2 birds until 45 wk of age. Egg production data were analyzed for hen-day production, laying sequence length, egg and egg component weights, time of lay, and egg interval time as related to strain or LI. Hen-day production was greater in brown egg strains (ISA B = 86.7%, S579 = 88.1%) than in white egg strains (ISA-W = 83.4%, S2000 = 82.3%) and was reduced in birds under 1 lx compared to 5 or 50 lx. A LI of 1 lx resulted in reduced egg production and laying sequence length compared to birds with a 50 or 500 lx. The effects of LI were strain dependant, however. Postpeak sequence length and egg production declined at more rapidly under 500 lx compared to other LI in brown egg strains, indicating possible development of a photorefractory condition. Mean settable egg weight was lower in 500 lx birds (56.1 g) compared to other groups (mean = 57.9 g), reducing total egg mass produced. Mean interval between successive eggs in a sequence was lengthened in 1 lx birds compared to other LI groups. Mean time of lay was earlier in brown egg strains than in white egg strains by 48 min. Mean time of lay was shifted to occur later by an increasing LI. Light intensity affected sexual maturation and egg production, as layers had differential responses to lighting. LI of 1 lx and 500 lx were found to be limiting to the egg production efficiency of layers. Whereas the birds receiving 1 lx had a reduced rate of egg production, those receiving 500 lx had reduced egg size later in the production period in combination with reduced shell quality, which indicated that inadequate feed intake under high LI conditions may be a factor affecting layer stocks. Exposure to high LI reduced egg size and total egg mass produced. Ultimately, the brown egg strains appeared to be more susceptible to the negative effects of low or high LI, indicating the importance of matching management practices to the particular hen genotype. PMID- 11495465 TI - An epidemiological study of the hatchability in broiler breeder flocks. AB - The aim of the present retrospective longitudinal study was to identify factors that significantly influenced the hatchability observed during the period from 1996 to 1998. Data from 112 Ross 208 breeder flocks in 39 breeding farms were included in a repeated-measures analysis with the flock-level "interval-specific" hatchability of the eggs as outcome variable. The overall mean of interval specific hatchability in the study was 75.9% (95% confidence interal: 75.3 to 76.6%). The hatchability was influenced by the storage time from lay until incubation and whether the farmers delivered floor eggs for hatching. The method used for disinfecting the hatching eggs and the flock average slaughter weight were significantly associated with the hatchability. A significant effect of time (production year) and flock size were also found on the hatching results. The results showed that several factors may influence the hatchability, the most detrimental being long egg storage time. With an optimization of the capacity and administration of the hatcheries, it would be possible to improve the hatching results. PMID- 11495466 TI - The effects of genetic selection on production parameters of single comb White Leghorn hens. AB - Four commercial table egg genetic stocks consisting of the Ottawa Control Strains 5, 7, and 10 (CS5, CS7, and CS10) and the 1993 H&N "Nick Chick" (CCS) were housed in the same environment and compared for production characteristics. These birds were housed in an environmentally controlled laying facility with trideck cages. Feed consumption, egg production, and mortality were monitored daily and compiled every 28 d. The study was conducted for two egg production cycles, including the molt period. Body weight was progressively lower for the more modern strains with CS5 being the heaviest and CCS maintaining the smallest body weight throughout the production periods. The CCS had the highest (P < 0.0001) hen-day production rate, which resulted in the greatest daily egg mass among the strains. The CCS consumed the greatest amount of feed and exhibited the highest gross egg income among the strains. We concluded that genetic selection has improved production parameters in commercial layers as determined by measurements in this study. PMID- 11495467 TI - Effect of level of acidification by phosphoric acid, storage temperature, and length of storage on the chemical and biological stability of ground poultry mortality carcasses. AB - Two experiments were conducted to evaluate the addition of feed-grade H3PO4 in comparison to lactic acid fermentation as a means of preserving ground poultry mortality carcasses. Mortality silage quality in both experiments was evaluated by measuring the rise in pH after initial acidification (deltapH) and the content of nonprotein N (NPN), volatile nitrogen (VN), NH3-N, and fecal coliform bacteria in the silage treatments. Preliminary work in Experiment 1 evaluated the preservative effects of six initial levels of acidification with H3PO4 (pH 2.0, 2.5, 3.0, 4.0, or 5.0) at two storage temperatures (21 and 45 C). Experiment 2 compared silages preserved with different levels of feed-grade 74.5% H3PO4 (4.13, 5.52, 6.90, and 8.28% wt/wt, concentrated acid basis) to silages made by lactic acid fermentation. Based on the evaluations in Experiment 1, acidification with H3PO4 to pH < or = 3.0, when incubated at 45 C, or pH < or = 2.0, when incubated at ambient temperatures, produced the most biologically favorable poultry mortality silage for subsequent use as a feed ingredient. In Experiment 2, mortality silages containing 8.28% H3PO4 (wt/wt, concentrated acid basis) contained significantly (P < 0.05) lower levels of the protein degradation by products VN and NH3-N than silages prepared by lactic acid fermentation. Therefore, feedstuffs manufactured from mortality silages prepared using 8.28% H3PO4 would be expected to result in improved animal performance vs. feedstuffs manufactured using silages prepared by lactic acid fermentation. PMID- 11495468 TI - The effect of ethoxyquin on the quality of ground poultry mortality carcasses preserved by lactic acid fermentation and phosphoric acid stabilization. AB - Fermentation and acidification have been shown to preserve the protein quality of ground poultry coproducts, but the effects of these processes on their lipid stability are unknown, especially in the presence of an antioxidant. To evaluate the effects of these treatments on lipid quality, ground poultry mortality carcasses, with and without an addition of 500 ppm ethoxyquin, were stabilized for 14 and 45 d by lactic acid fermentation or acidification with 2.76, 5.07, 7.35, or 9.65% feed-grade H3PO4. Ethoxyquin treatment significantly (P < 0.001) improved the oxidative stability of lipids from all storage treatments. However, the addition of ethoxyquin increased (P < 0.001) the levels of volatile N (VN) from 2.51 to 3.18% in products stored for 45 d and resulted in an increase (P < 0.001) in free fatty acids in all ensiled products. Ethoxyquin addition had no effect (P > 0.120) on the fatty acid profile of products stored for 14 d but significantly increased (P < 0.001) the levels of stearic (C18:0) and arachidonic acids (C20:4) in products stored for 45 d. In this experiment, the addition of ethoxyquin to preservation systems for the short-term storage of poultry mortality carcasses improved the lipid quality of the ground material without compromising the protein quality or affecting proximate analysis parameters. However, the increased oxidative stability of mortality silage materials that contain ethoxyquin may contribute to enhanced microbial or enzymatic activities that result in proteolytic or lypolytic breakdown products following longer periods of storage. PMID- 11495469 TI - Effects of dietary polyunsaturated fatty acids on in vivo splenic cytokine mRNA expression in layer chicks immunized with Salmonella typhimurium lipopolysaccharide. AB - Effects of dietary polyunsaturated fatty acids (PUFA) on immune responses in poultry have been reported. However, effects on the underlying mechanisms, such as the role of cytokines, have not been documented because the necessary tools were lacking. Recently, primer sets for chicken interleukin (IL)-1beta, IL-2, interferon-gamma (IFN-gamma), myelomonocytic growth factor (MGF), and transforming growth factor (TGF)-beta2 have become available. Therefore, in the present study we first examined the in vivo effects of an inflammatory challenge with Salmonella typhimurium lipopolysaccharide (LPS) on cytokine profiles in growing laying-type chicks. Second, we examined whether dietary fat sources affected the observed cytokine profiles. Two hundred forty chicks were assigned in a 2 x 4 factorial design of treatments, with injection with LPS or saline and dietary fat source as factors. Factors were i.v. injection with S. typhimurium LPS or saline (control) and four dietary fat sources: corn oil, linseed oil, menhaden oil, and tallow. Two hours after injection, birds were killed, and their spleens were removed for RNA extraction. Reverse transcription polymerase chain reactions with primer sets for chicken IL-1beta, IL-2, IFN-gamma, MGF, TGF-beta2, and beta-actin were performed with RNA samples pooled by pen. The expression of cytokine mRNA was expressed relative to the level of beta-actin mRNA. Interleukin 1 (P < 0.001), MGF (P < 0.0001), IL-2 (P < 0.001), and IFN-gamma (P < 0.001) mRNA expressions were enhanced by challenge with LPS. Immunization treatment had no effect on TGF-beta2 or beta-actin expression. Dietary treatment did not affect mRNA expression of IL-1, MGF, IFN-gamma, TGF-beta2, or beta-actin. Interleukin-2 expression in LPS-injected birds that were fed the fish-oil-enriched diet was enhanced (P = 0.05). The present study indicates that in vivo effects of immune challenge on cytokine mRNA expression can be measured in poultry. The observation that mRNA level of IL-2, but not the mRNA levels of IFN-gamma or MGF, is enhanced by dietary fish oil at 2 h suggests that dietary PUFA at this moment initially affected naive T lymphocytes. PMID- 11495470 TI - Influence of source and percentage of fat added to diet on performance and fatty acid composition of egg yolks of two strains of laying hens. AB - We conducted a trial to study the influence of strain of hen (white vs. brown) and source and percentage of added fat to diet on productive performance and fatty acid (FA) profile of the egg yolk. There were nine dietary treatments: a control diet without added fat and eight diets arranged as a 2 x 4 factorial (5 vs. 10% added tallow, olive, soy, or linseed oil). Each treatment was replicated five times, and the trial lasted 12 wk. White hens produced yolks with more saturated FA (P < 0.001) and less monounsaturated FA (P < 0.001) and polyunsaturated FA (P < 0.05) than brown hens. Fat supplementation improved egg mass output (P < 0.05) and feed efficiency of hens (P < 0.001). Linseed and soy oil diets produced greater egg mass than diets based on tallow or olive oil (59.6, 59.4, 57.9, and 57.1%, respectively; P < 0.001). The proportion of saturated FA in the yolk decreased as the percentage of added fat increased (36.5, 32.3, and 31.0% for diets containing 0, 5, and 10% added fat, respectively; P < 0.001). When the alpha-linolenic acid content of the diets increased from 0 to 0.8%, the arachidonic, docosapentanoic, and docosahexanoic acids in the egg yolk increased. Increases in the alpha-linolenic acid content of the diet over 2.3% did not further augment the concentration of docosapentanoic or docosahexanoic acid of the yolk and, in fact, decreased that of arachidonic acid. Only diets supplemented with linseed oil produced eggs with measurable amounts of eicosapentanoic acid. Diets supplemented with soy oil, with moderate levels of alpha-linolenic acid and high levels of linoleic acid, increased arachidonic and docosahexanoic acids in the egg yolk but not eicosapentanoic acid. PMID- 11495471 TI - Nutritive value of the meat and bone meals from cattle or pigs in broiler diets. AB - The nutritive value of meat and bone meals (MBM) was assessed for broilers. The MBM was produced according to the revised (pressure) processing system ordered by the European Union (EC 96/449). Three batches of MBM from cattle (MBMcattle) and three from pigs (MBMpig) with different ash contents (224, 306, 387, and 209,293, 430 g/kg, respectively) were tested for digestibility at a 10% inclusion level. The MBMcattle and MBMpig with the lowest ash (224 and 209 g/kg, respectively) were tested also at 20% inclusion. A basal diet (corn-soybean meal) was used as a control. Two-week-old broiler chickens were used in four replicates per treatment (14 to 32 d of age). The AMEn of MBM was high (10.51 to 13.04 MJ/kg DM). Species origin had no significant effect, whereas more ash and a higher inclusion level decreased the AMEn. The factors investigated showed no significant effect on the excretal digestibility of CP or on total AA. Excretal digestibility of total amino acids (AA) ranged from 60 to 65%. The ileal digestibility of CP and AA of MBMpig with 209 g/kg ash was also tested at 10 and 20% inclusion. Excretal digestibility was significantly higher than ileal digestibility of CP (63.8 and 55.8%, respectively) and total AA (60.9 and 56.2%, respectively). The 20% inclusion level resulted in a lower digestibility for both methods. The digestibility of CP was measured by four different in vitro techniques, based on pepsin digestibility. The data showed a large variation and did not correlate at all with the in vivo digestibility values. PMID- 11495472 TI - Effects of vitamin E and C supplementation on performance, in vitro lymphocyte proliferation, and antioxidant status of laying hens during heat stress. AB - Vitamin E (dl-alpha-tocopheryl acetate) was evaluated for its effects on performance, lymphocyte proliferation, and antioxidation in layers during heat stress. In Trial 1, 25, 45, or 65 IU of vitamin E/kg were fed to four replicated pens (five hens/cage) of DeKalb Delta or Hy-Line W-36 per treatment starting at 20 wk of age. At 34 wk of age, hens were heat-stressed at diurnal temperature ranging from 21 C to 35 C for 3 wk. The performances of hens not exposed to heat stress were not influenced by supplemental vitamin E. Supplemental vitamin E did not affect egg production; however, egg mass was greater (P < 0.05) with supplementation of 65 IU of vitamin E/ kg during heat stress. Egg yolk was significantly increased (P < 0.04) when hens were fed 45 and 65 lU/kg compared with the control vitamin E level (25 lU/kg). Haugh units were higher (P < 0.01) for hens fed 65 IU of vitamin E/kg compared to 25 and 45 lU/kg. Lymphocyte proliferative responses to concanavalin A (Con A) and Salmonella typhimurium lipopolysaccharide (LPS) were greater (P < 0.0001) in hens fed 45 and 65 IU of vitamin E/kg during heat stress. Strain had no effect on any of the parameters measured. In Trial 2, a 2 x 2 factorial was designed to test effects of vitamin C in drinking water (0 and 1,000 ppm) and dietary vitamin E (25 and 65 IU/kg). Eight replications per treatment with four hens per replication cage were heat stressed at constant temperature of 35 C for 3 wk. Egg production and egg mass were higher when hens were fed 65 IU of vitamin E/kg than when hens were fed 25 lU/kg (81.5 vs. 75.9%, P < 0.03 and 48.2 vs. 44.6 g, P < 0.03, respectively). Yolk solids weight for the 65 IU vitamin E/kg group was higher (P < 0.01) compared to the 25 IU/kg group. ConA and LPS mitogenic responses were greater in hens fed 65 IU of vitamin E (P < 0.001 or P < 0.003, respectively) or 1,000 ppm of vitamin C (P < 0.001 or P < 0.002, respectively). The combination of 65 IU vitamin E/kg and 1,000 ppm vitamin C showed the highest ConA and LPS mitogenic responses among the treatments. No interaction effects of the two vitamins on production measurements or lymphocyte proliferative responses were observed. TBA values in egg yolk and plasma of hens fed 65 IU of vitamin E/kg were lower (P < 0.0001) than those of hens that received 25 IU of vitamin E/kg. These results suggest that vitamin E supplementation at 65 IU/kg diet may enhance production, induction of in vitro lymphocyte proliferation by ConA and LPS, and antioxidant properties of egg yolks and plasma of White Leghorn hens during heat stress and that supplementation of 1,000 ppm vitamin C may further enhance in vitro lymphocyte proliferative responses of hens during heat stress. PMID- 11495473 TI - Dietary effects of soy isoflavones on growth and carcass traits of commercial broilers. AB - Three experiments (EXP) were conducted to determine the effect of soy isoflavones (ISF) on growth and carcass traits of commercial broilers. The EXP were conducted simultaneously and a common control was used. In each EXP, treatments were replicated five times with five chicks each. Average initial and final BW were 102 and 2,890 g, 102 and 2,657 g, and 102 and 2,803 g for EXP 1,2, and 3, respectively, and the EXP were conducted from 9 to 52 d posthatching. In EXP 1, the effects of graded levels of supplemental ISF above those typically found in a corn-soybean meal (C-SBM) diet were studied. The treatments were 1) C-SBM, 2) C SBM + ISF levels two times those in C-SBM (2x), and 3) C-SBM + ISF levels five times those in C-SBM (5x). The 2x and 5x levels of ISF decreased gain:feed (linear, P < 0.04) but increased (P < 0.04) breast weight compared with the C-SBM diet. Other performance and carcass traits were not affected (P > 0.05) by treatment. In EXP 2, the effects of low ISF levels in a C-soy protein concentrate (C-SPC) diet were studied. The treatments were 1) C-SBM, 2) C-SPC (low ISF), and 3) C-SPC + ISF (ISF levels equal to those in C-SBM). Average daily gain (ADG) and average daily feed intake (ADFI) were decreased (16 and 9%, respectively; P < 0.01) in chicks fed the C-SPC diets, regardless of ISF level. Gain:feed of chicks fed the C-SPC + ISF diet was decreased 9% (P < 0.02) compared with chicks fed the C-SBM diet, and gain:feed of chicks fed C-SPC was intermediate between the two. Carcass traits were not affected (P > 0.05) by treatment. In EXP 3, the effects of low ISF levels in a low CP diet were studied. The treatments were 1) C-SBM, 2) low CP (17,14, and 12% in the starter, growing, and finishing diets, respectively) with supplemental crystalline amino acids (low CP-AA), and 3) low CP-AA + ISF (ISF levels equal to C-SBM). Daily gain and gain:feed were decreased from 7 to 9% (P < 0.01) in chicks fed the low CP-AA and low CP-AA + ISF diet relative to those fed the C-SBM diet. Abdominal fat pad percentage was increased (P < 0.01) in chicks fed the low CP-AA diets compared with those fed the C-SBM diet. Dietary ISF can affect ADG and ADFI and may affect carcass traits in some instances. PMID- 11495474 TI - Effects of dietary 1, 4-diaminobutane (putrescine) on eggshell quality and laying performance of older hens. AB - We conducted an experiment to evaluate the potential for dietary 1,4 diaminobutane (putrescine) to promote eggshell quality and overall laying hen performance. A total of 128, 60-wk-old Barred Rock hens were fed a corn and soybean meal-based layer diet supplemented with 0.0 (control), 0.2, 0.4, and 0.6% free base 1,4-diaminobutane for 4 wk. The feeding of supplemental putrescine decreased feed consumption (P < or = 0.05) and egg mass (P < or = 0.05) and tended to decrease egg production (P < 0.08). Albumen quality was not significantly affected (P < 0.09) by the end of the experiment, as determined by Haugh units. Eggshell thickness was not significantly improved with lower levels of dietary putrescine (P < 0.08). Although dietary putrescine did not have any effect on the relative weights of duodenum, jejunum + ileum, or pancreas, there was a linear increase in putrescine concentrations in tissues (P < or = 0.05). Supplementation of dietary putrescine also resulted in increased putrescine and spermidine concentrations in egg (P < or = 0.05). Egg weight and eggshell deformation increased over time; however, eggshell weight, eggshell weight as percentage of egg weight, and eggshell thickness decreased (P < or = 0.05). It appeared that eggshell quality declined regardless of diet over the 4-wk experimental period. It was concluded that the lack of effect of dietary putrescine on egg parameters, with the exception of albumen quality and eggshell thickness, was due to putrescine toxicity. Hens transferred excess dietary putrescine and metabolites to eggs. PMID- 11495475 TI - 1. Appetite suppressant activity of supplemental dietary amino acids and subsequent compensatory growth of broilers. AB - This study was conducted to take advantage of the appetite-suppressant effect of excessive dietary amino acids in reducing feed intake and, in turn, restricting the early rapid growth of broilers to minimize metabolic disorders. Dietary amino acids were supplemented to a basal diet to yield a total of 1.57, 2.57, and 3.57% His; 2.7, 4.3, and 5.9% Lys; 1.36, 2.16, and 2.96% Met; 2.8, 3.8, and 4.8% Thr; and 1.27, 2.27, and 3.27% Trp and were fed to 408 chicks from 4 to 11 d of age. Fifteen dietary treatments of His, Lys, Met, Thr, and Trp were compared to the basal diet. Feed consumption was measured daily. Body weight measurements were taken at 0, 4, 7, 11, 14, and 21 d. At 21 d, pectoralis major and minor muscles, liver, and abdominal fat pad were weighed. High levels of Met and His caused the greatest depression in appetite from 4 to 11 d, and Thr, Trp, and Lys were found to be less potent. The exponential growth rate (EGR) of birds from 4 to 11 d of age was significantly reduced by the intermediate and high levels of the amino acid supplementation. From 11 to 14 d, EGR was greatest with high levels of Met or Trp, indicating more potential compensatory growth realized with these treatments. The high level of His decreased the percentage of pectoralis minor muscle yield, whereas the high level of Lys and Met increased the percentage of liver compared to those fed the basal diet. These results indicate that it is possible to use excessive individual amino acids in diets to suppress the appetite and early rapid growth to alleviate or minimize metabolic disorders. PMID- 11495476 TI - Growth and digestive function of turkeys surviving the poult enteritis and mortality syndrome. AB - Stunted growth of poults afflicted by enteritis mortality syndrome (PEMS) may be associated with depressed digestive capabilities. We conducted two experiments to test this hypothesis. Survivors of PEMS were obtained from a research flock that had been purposefully infected at 5 d of age with PEMS inoculum that included turkey corona virus. The PEMS survivors were assigned to Experiments 1 and 2, beginning at 40 and 35 d of age, respectively. Three groups (large, L; medium, M; and small, S) and two groups (L and S) of 20 poults each were selected to represent different degrees of stunting in Experiments 1 and 2, respectively. When the body weights of each group in both experiments were plotted using initial body weight as the starting point, all of the weights fell on the normal growth curve except Group S in Experiment 1. Therefore, there was no evidence of compensatory growth over the period studied. In Experiment 1, apparent fat absorption (AFA) was significantly higher (P < 0.05) for control (85.9%) than for M (78.5%) and S (78.3%) groups, and AME, for the control group was significantly higher than all the PEMS-survivor groups. In Experiment 2, Group L had a higher AFA and AMEn than Group S. AFA and AMEn improved in both groups over time. The digestive capabilities of PEMS survivors were depressed proportionally to the degree of stunting. Impaired fat digestibility and dietary energy utilization in PEMS-afflicted birds are likely contributors to stunted growth and reduced recovery rates. PMID- 11495477 TI - Developmental changes of parathyroid hormone/parathyroid hormone-related peptide and calcitonin receptor binding properties in the chicken calvaria and kidney. AB - The equilibrium dissociation constant (Kd) and the maximum binding capacity (Bmax) of receptors for parathyroid hormone (PTH)/parathyroid hormone-related peptide (PTHrP) and calcitonin (CT) in the membrane fraction of the calvaria and the kidney of chickens were examined from 1 d after the hatch up to 24 wk of age by the use of radioligand binding assays. The Kd values of the PTH/PTHrP receptor in both tissues were decreased at 10 and 24 wk in female birds, whereas the values were increased at 24 wk in male birds. The Bmax values of the PTH/PTHrP receptor in both tissues were decreased at 10 wk and returned to baseline at 24 wk in female birds. The values were increased at 24 wk in male birds. The Kd and Bmax values of CT receptors in the both tissues were constant during the experimental period in female and male birds. The results suggest that the binding properties of PTH/PTHrP receptor and of CT receptor may be influenced by gonadal hormones relating to sexual maturation. PMID- 11495478 TI - The effect of acidified soapstocks on feed conversion and broiler skin pigmentation. AB - The effect of different soapstocks (corn, sunflower, canola, and soybean) on productive performance and skin broiler pigmentation was investigated. Soapstock was added to reach 1.0% polyunsaturated fatty acids in the diet. The addition of soybean soapstock significantly improved live body weight gain of the birds from 1 to 7 wk of age. A live body weight gain of 1,736 g/bird was calculated for broilers fed with the soybean soapstock diet. Feed conversion was significantly higher for broilers fed with the soybean soapstock diet, and no negative effect was observed. Compared to broilers fed with Pixtafil (100.0% pigmentation), those fed soybean soapstock (when added as a supplement of 1.0% polyunsaturated fatty acids in the diet) reached 48.0% pigmentation, and those fed corn soapstock reached only 7.3%. When the diets were complemented with Pixtafil to reach 100% of calculated pigmentation, the soybean soapstock diet reached 100.8% pigmentation compared to a canola soapstock diet that reached 72.0% pigmentation. Acidified soybean soapstock could be a source of polyunsaturated fatty acids and of xantophyl pigments in broiler feeding. PMID- 11495479 TI - Effect of storage and layer age on quality of eggs from two lines of hens. AB - Eggs from ISA-White and ISA-Brown hens between 28 and 59 wk of age were stored for up to 10 d to produce a sample of 5,763 eggs differing in the three major determinants of albumen quality. Eggs from ISA-Brown hens were larger and had less yolk, more albumen, and a greater percentage of shell than those from ISA White hens. Egg size increased with increasing age of the hen, although more for the ISA-White hens than the ISA-Brown hens, and the yolk increased more in size than did the shell and albumen. During storage, albumen weight decreased and yolk weight increased slightly. The height of the inner thick albumen of eggs from ISA White hens was greater than that of eggs from ISA-Brown hens, and it decreased as the hen age increased and with increasing time in storage. The pH of the albumen was not different between strains, and the effect of hen age was small, but it increased with time in storage. Regression coefficients of the height of the inner thick albumen on the weight of the egg were between -0.058 and 0.102, showing that the fixed regression of 0.05-mm albumen height per gram of egg implied by the Haugh unit is wrong. The statistical association between albumen pH and egg weight was very low. If albumen quality is being used as a measure of freshness, then the albumen height is biased by the strain and age of hen, whereas the albumen pH is not. PMID- 11495480 TI - Impulse oscillometry in healthy nonsmokers and asymptomatic smokers: effects of bronchial challenge with methacholine. AB - The clinical application of respiratory impedance measurements by oscillation techniques for monitoring bronchial challenge testing is hampered by the fact that data in healthy nonsmokers and asymptomatic smokers are very limited. The objective of this study was to analyze the changes in impedance to a methacholine provocation test in healthy nonsmokers and asymptomatic smokers, and to investigate whether smokers show a different response compared to nonsmokers. The response to methacholine challenge was assessed by impulse oscillometry (IOS) (resistance R and reactance X at 5, 10, 15, 20, 25, and 35 Hz) and spirometry (FEV1, MEF50) in 105 healthy subjects (55 nonsmokers: "NS"; 50 asymptomatic smokers: "S") in whom the provocation dose of 2.44 mg methacholine did not result in a fall of FEV1 below 20% predicted of the baseline value. The baseline reactance X values of S were significantly lower compared to NS from 10 to 35 Hz (p < or = 0.01), whereas S and NS did not differ in resistance R over the whole frequency spectrum from 5 to 35 Hz. S showed a significantly higher mean baseline resonant frequency X(f0); i.e., the frequency at which inertial forces are equal and opposite to elastic forces, compared to NS (NS: X(fo) = 10.8+/-2.9 Hz; S = 12.6+/-4.0 Hz, p = 0.01). In both groups methacholine challenge resulted in an increase in R (f) especially at 5 and 10 Hz, and a marked decrease in X(f) over the whole frequency spectrum. In S a significantly higher decrease of postchallenge X5Hz and X10Hz was measured compared to NS (NS: deltaX(5Hz) = 0.044+/-0.003; S: deltaX(5HZ) = -0.083+/-0.013; p = 0.0017. NS: deltaX(10Hz) = 0.050+/-0.001; S: deltaX(10Hz) = -0.082+/-0.017; p = 0.008). R and X at low frequencies showed a three to four times higher postchallenge reaction compared to FEV1. Pre- and postchallenge FEV1 did not correlate with resistance R (f) and reactance X(f) over the whole frequency spectrum. In S perchallenge X(f) values from 10 to 35 HZ were significantly lower, and postchallenge decrease of X5Hz and X10Hz values were significantly higher compared to that of NS, whereas pre- and postchallenge R(f) values were similar in both groups. These results can be explained by changes in the elastic properties of the lung induced by a diffuse subclinical respiratory bronchiolitis. PMID- 11495481 TI - Validation of laser diffraction method as a substitute for cascade impaction in the European Project for a Nebulizer Standard. AB - The project for a European standard testing procedure to characterize nebulizers in terms of particle size distribution has been based on using the Andersen Marple personal cascade impactor model 298 (A-MPCI) with a sodium fluoride reference solution. In the present study methods based on laser diffraction (Mastersizer-X) and time-of-flight (TOF)(APS) and another cascade impactor (GS1 CI) were compared with the A-MPCI. Two types of nebulizer (Pari LC+ and Microneb) were tested with all apparatuses, and a third type of nebulizer (NL9) was tested with the A-MPCI and Mastersizer-X. Nebulizers were charged with a solution of sodium fluoride in conditions reproducing the European Committee for Normalization (CEN) protocol. There was no difference between the Mastersizer-X and the A-MPCI or between the GS1-CI and the A-MPCI in terms of mass median aerodynamic diameter (MMAD). Comparison between the APS and the A-MPCI showed a significant difference with the Microneb. The geometric standard deviations (GSD) obtained with the A-MPCI were on average 10% greater than GSD obtained with the other apparatuses, but the differences were not statistically significant. We conclude that laser diffraction can be used for particle size distribution in the context of the European standard, and that the Mastersizer-X is particularly interesting for industrial practice in view of its simplicity and robustness. PMID- 11495482 TI - Effect of particle inlet distributions on deposition in a triple bifurcation lung airway model. AB - Considering a triple bifurcation as a representative lung airway model of the upper bronchial tree, the effect of different random inlet particle distributions on deposition patterns and efficiencies have been numerically analyzed. The steady laminar three-dimensional transport equations for a dilute micron-size particle suspension have been solved using a commercial finite-volume code with user-enhanced programs. Particle release positions were assigned employing a random number generator following random-parabolic, random-uniform, and random random distribution functions. Via back tracking, starting positions of all depositing particles were determined for each particle release distribution, including a deterministic-parabolic one which served as a base case. The results indicate that: (1) The starting regions of the depositing particles in a given bifurcation are fixed for the same inlet Reynolds number and Stokes number combination, regardless of the type of distribution profile. The situation for the particle deposition patterns is somewhat similar. However, the type of distribution of inlet particles strongly influences the particle deposition efficiencies. (2) Values of particle deposition efficiencies are very close for the same (parabolically) distributed deterministic versus random inlet particles when all other conditions are fixed. (3) According to the simulation validations, a determinstic parabolic distribution of inlet particles may be sufficient for laboratory data comparison purposes, but random distributions should reflect realistic environmental or medical aerosol inhalation more accurately. PMID- 11495483 TI - The equivalence of compressor pressure-flow relationships with respect to jet nebulizer aerosolization characteristics. AB - Manufacturers of aerosolized medications, approved by the Food and Drug Administration, specify the nebulizer(s) and compressor to be used with their product, in an attempt to achieve efficacy comparable to that obtained in the clinical trials. The need to limit the compressor to that used in the trials has not been investigated in detail. We suggest a technique to determine the equivalency of different compressors such that a chosen nebulizer's performance is not significantly altered. Aerosol particle size (MMD) was measured with a laser; compressor flow and pressure were measured with a mass flow meter and pressure gauge, respectively. For all models of nebulizer, increased flow or driving pressure caused a decrease in aerosol MMD. The flow resistance of nebulizer models varied, and the flow output of compressors decreased as imposed nebulizer resistance increased. However, for any specific compressor-nebulizer combination there is a unique flow and pressure, and the nebulizer generates a given MMD. We demonstrate methods to choose alternate compressors that may be used to drive a nebulizer and yet keep the nebulizer's MMD and performance within predetermined limits. Once an acceptable range of variance in a nebulizer's MMD is defined, alternate compressors may be safely chosen. We recommend that these techniques be used by manufacturers of medications and of compressors to safely determine the acceptability of several rather than a single model compressor to drive a chosen nebulizer. The techniques assure consistency of the nebulizer's clinically demonstrated performance characteristics. PMID- 11495484 TI - Performance of heated humidifiers with a heated wire according to ventilatory settings. AB - Delivering warm, humidified gas to patients is important during mechanical ventilation. Heated humidifiers are effective and popular. The humidifying efficiency is influenced not only by performance and settings of the devices but the settings of ventilator. We compared the efficiency of humidifying devices with a heated wire and servo-controlled function under a variety of ventilator settings. A bench study was done with a TTL model lung. The study took place in the laboratory of the University Hospital, Osaka, Japan. Four devices (MR290 with MR730, MR310 with MR730; both Fisher & Paykel, ConchaTherm IV; Hudson RCI, and HummaxII; METRAN) were tested. Hummax II has been developed recently, and it consists of a heated wire and polyethylene microporous hollow fiber. Both wire and fiber were put inside of an inspiratory circuit, and water vapor is delivered throughout the circuit. The Servo 300 was connected to the TTL with a standard ventilator circuit. The ventilator settings were as follows; minute ventilation (V(E)) 5, 10, and 15 L/min, a respiratory rate of 10 breaths/min, I:E ratio 1:1, 1:2, and 1:4, and no applied PEEP. Humidifying devices were set to maintain the temperature of airway opening at 32 degrees C and 37 degrees C. The greater V(E) the lower the humidity with all devices except Hummax II. Hummax II delivered 100% relative humidity at all ventilator and humidifier settings. When airway temperature control of the devices was set at 32 degrees C, the ConchaTherm IV did not deliver 30 mg/L of vapor, which is the value recommended by American National Standards at all V(E) settings. At 10 and 15 L/min of V(E) settings MR310 with MR730 did not deliver recommended vapor, either. In conclusion, airway temperature setting of the humidifying devices influenced the humidity of inspiratory gas greatly. Ventilatory settings also influenced the humidity of inspiratory gas. The Hummax II delivered sufficient water vapor under a variety of minute ventilation. PMID- 11495485 TI - What is the GAT? Aerosoltherapy Group. PMID- 11495486 TI - Aerosols and anti-infectious agents. AB - Anti-infectious agents such as pentamidine, antibiotics (mainly colistine and aminoglycosides), and amphotericin B can be administered by aerosol. Apart from pentamidine and Tobi, this route of administration is not officially approved and it constitutes an empirical approach, which has benefited from recent research summarized hereafter. The most fundamental question is related to the potentially deleterious effects of nebulization processes, especially ultrasound, on the anti infectious properties of the drugs. Colimycin, which was chosen as a reference because its polypeptide structure makes it unstable a priori, proved to be resistant to high frequency ultrasound, which is encouraging for other molecules such as aminoglycosides or betalactamins. The nebulizer characteristics also have to be taken into account. An aerosol can be produced from an amphotericin B suspension and from colistine using both an ultrasonic nebulizer and a jet nebulizer. Differentiating between good and bad nebulizers is not dependent upon the physical process involved to nebulize the drug, but on the intrinsic characteristics of the device and its performance with a known drug. The inhaled mass of an aerosol in the respirable range must be high and dosimetric nebulizers represent significant progress. Finally, administration of anti-infectious aerosols requires a new pharmacological approach to monitor treatment, and urinary assays are promising for this purpose. PMID- 11495487 TI - A history of nebulization. AB - The simplest and most natural route of drug delivery to the lungs is the inhaled one. From the historical and medical point of view, it was a Greek, Pedanus Discorides, the father of the science of pharmacy, who, during the first century prescribed inhaled fumigation. Pipes were also used to inhale hallucinogenic substances. All shamans knew the psychotropic effects of poisonous plants such as Datura stramonium, especially Red Indians, in their peace calumets; but Indians of Madras used fumigations of Datura ferox to treat asthma. Since 1803, this therapeutic was imported in Great Britain and cigarettes with leaves of datura were used by asthmatics until 1992. In the middle of the nineteenth century, to treat grapevines diseases and in response to the fashion of inhaling thermal waters, spray technology was developed for the effervescent waters at the thermal spas. The onslaught of tuberculosis, similar to AIDS a century later, brought back into practice the inefficacious use of antiseptic aerosol therapy. With the discovery of adrenaline, ephedrine aerosols enjoyed a rebirth. The perfecting of jet nebulizers by R. Tiffeneau, father of FEV1 and M.B. Wright, father of peak flow, allowed a better practice of inhalotherapy. In 1949, the United States, ultrasonic nebulizers made their first appearance in the form of humidifiers, but doctors were quick to add medications to produce therapeutic aerosols. After 150 years, with the improvement of nebulizer systems and new nebulized medications, the nebulization story is still not concluded. PMID- 11495488 TI - A simple tool for monitoring nebulized amikacin treatments based on a single urine assay. AB - Aerosolized aminoglycosides have demonstrated their efficacy in the treatment of P. aeruginosa pneumonia in cystic fibrosis (CF) patients. There is wide interpatient variability in the deposited and systemic drug doses that depend on both the nebulization and inhalation conditions and result in a risk of inefficacy or toxicity. We have developed a tool to provide a simple method for individual dose monitoring by estimating the total quantity of amikacin excreted, which corresponds to the dose absorbed systemically. It is based on a single urine assay. Thirty-seven urinary pharmacokinetic time courses in healthy volunteers (groups A and B) or in CF patients (groups C and D) were used. The rules for extrapolating the total dose excreted on the basis of 6-, 8-, 10-, and 12-h urine samples, were determined from group A. The accuracy of these rules was then tested in the other three groups. The total amount excreted was poorly predictable, with a coefficient of variation (CV) of 36 and 30% in the healthy volunteers, and of 48 and 82% in the CF group, whereas the CV of the estimated amount, based on 8- to 12-h samples, was only 10-15% in the healthy volunteers and 4-8% in the CF patients. Collecting a single sample over an 8- to 12-h period requires overnight sampling. The very low circadian variations in renal function, ranging from -2% to +5%, demonstrated the absence of any significant bias resulting from overnight sampling. A single urine assay can therefore be proposed as a simple, noninvasive, low cost, and reliable method for the clinical monitoring of nebulized amikacin in CF patients. Further studies are needed before this method can be extended to aerosol treatments with other aminoglycosides. PMID- 11495489 TI - Preliminary study of the deposition of aerosol in the maxillary sinuses using a plastinated model. AB - In spite of the widespread use of aerosols in respiratory diseases, very few studies have been performed in the field of ear, nose, and throat (ENT) disorders. The conditions for penetration of aerosols inside the sinus cavities are thus still not understood fully. The aim of this study was to investigate the penetration of aerosols inside maxillary sinuses in vitro, using plastinated models. Three plastinated specimens of the nose and sinuses were made from three different corpses. These specimens were validated by CT scans and were used to study deposition of aerosol in the maxillary sinuses. We performed scintigraphic images of the models in above, face, and profile views using a technetium (99mTc) labelled solution to show aerosol deposition. We also counted the radioactivity deposited on gauze compresses placed inside the maxillary sinuses. In addition, we constructed a measuring unit with miniature humidity sensors placed inside the sinuses. We recorded the changes in relative humidity observed during nebulization. Results from these studies showed that scintigraphic images of the specimen, whatever the incidence of the views, were not accurate enough to differentiate the aerosol deposition in the maxillary sinuses from that in the nasal cavity. Using indirect counting on gauze compresses made possible the quantification of local aerosol deposition, and we found that aerosols entered into the sinuses. This confirmed that aerosols could reach the middle meatus, which is the main area for sinusitis disorders. The increased activity compared to background varied from 17 to 127%. The humidity sensors recorded changes in relative humidity during the nebulization. These humidity changes fitted a nonlinear model represented by the equation: y = b0 (1 - e(-b1t)), where b0 is the plateau and b1 is the speed to reach the plateau. These techniques may be useful in the future for in vitro characterization of aerosol penetration into the maxillary sinuses. PMID- 11495490 TI - Gene therapy for cystic fibrosis with aerosolized adenovirus-CFTR: characterization of the aerosol and scintigraphic determination of lung deposition in baboons. AB - For cystic fibrosis (CF) gene therapy using an aerosolized adenovirus expressing the CFTR gene, optimization of the inhalation conditions is a prerequisite to obtain sufficient amount of CFTR protein expression in the target areas of the respiratory tract. For such a purpose, in vivo radioisotopic imaging of the radiolabeled virus is a unique strategy for a quantitative assessment of the actual deposition. In the present study, an adenovirus CFTR (AdCFTR) was labeled with 99m Technetium gamma emitting isotope in such conditions that its bioactivity was preserved. The 99mTc-AdCFTR aerosol was characterized using both laser diffraction and cascade impaction for sizing with further determination of nebulized and inhalable fractions. After administration to baboons, scintigraphic quantitation of the regional lung distribution was performed and the actual dose deposited in the target area was estimated and expressed as an equivalent viral titer. Since a virus scintigraphy is not realistic in a hospital setting, we have developed an approach using 99mTc-DTPA (diethylene triamino pentaacetic acid) that could be used to predict the virus deposition. Indeed, similarities observed between 99mTc-DTPA and 99mTc-adenovirus aerosol imaging patterns validates the use of the 99mTc-DTPA scintigraphy that we propose as a pretherapeutic test for each patient prior to gene transfer. PMID- 11495491 TI - Assemblages of animals around urban structures: testing hypotheses of patterns in sediments under boat-mooring pontoons. AB - Assemblages of animals in soft-sediments were studied in relation to pontoons for mooring private boats in two estuaries near Sydney, Australia. Based on previously observed patterns around other types of artificial structures, it was predicted that assemblages of animals under pontoons would be different from those in similar areas away from pontoons. Hypotheses about overall differences in average abundance and composition between sites with and without pontoons were tested, as were hypotheses about variable differences among and within estuaries. Analyses revealed that there were fewer crustaceans under pontoons in one estuary. The most conspicuous patterns related to pontoons were, however, differences in variability among sites with pontoons compared to sites without pontoons. Differences in spatial variability were found for the overall multivariate structure using Bray-Curtis dissimilarities and for abundances of most major taxa. Total abundance was approximately 60 times more variable among sites without pontoons and number of taxa were seven times more variable among sites with pontoons. Such patterns indicate that impacts of pontoons occur at some sites but not at others. This may be explained by intrinsic differences among sites or by differences in practices for maintenance. Predictions from these two contrasting models need to be tested in order to achieve efficient management of this type of structure. PMID- 11495492 TI - Colonization of mine tailings by marine invertebrates. AB - An experiment was conducted to determine if a tailings substrate would inhibit recolonization of benthic macrofauna upon closure of a submarine tailings disposal (STD) operation. Trays of defaunated marine sediment, serving as a reference, and trays of tailings from a proposed gold mine were placed at 21 m depth on the ocean floor to allow colonization via settlement from the water column. Trays of reference sediment and tailings, and cores of ambient sediment were collected 9, 17, and 22 months after tray placement. Probable tray effects, which were desirable given the objectives of this study, precluded direct comparison of ecological succession in the tray sediments to the ambient assemblage. The ambient macrofauna assemblage was distinguishable from the reference sediment and tailings assemblages throughout the experiment and displayed more pronounced seasonality. Differences between the reference sediment and tailings assemblages were generally insignificant, including total abundance, total biomass, number of taxa, average size of individuals, numerically dominant taxa, abundance by ecological guilds, and overall community composition. Upon cessation of STD, recolonization of a benthic macrofauna community should not be inhibited by the presence of these tailings as a benthic substrate. PMID- 11495493 TI - A field-based population model for the sediment toxicity test organism Leptocheirus plumulosus: I. Model development. AB - We describe the development of a stage-structured population matrix model for the estuarine amphipod Leptocheirus plumulosus, a sediment toxicity test organism, based on regular sampling of a natural population for approximately 2 years. Annual population dynamics were characterized by abundance peaks in the spring and fall, and low densities in the summer and winter. We use a sequence of seasonal projection matrices to capture these qualitative patterns in population dynamics. We compare two methods for estimating transition probabilities: multiple regression and truncated singular value decomposition (SVD). Overall, SVD gave more realistic parameter estimates than regression, although fecundity estimation was problematic. There were predictable trends in growth, survival and fecundity within years. The SVD-derived model provides a framework for assessing the population-level effects of contaminants on Leptocheirus. PMID- 11495494 TI - A field-based population model for the sediment toxicity test organism Leptocheirus plumulosus: II. Model application. AB - A stage-structured population model has been developed for the estuarine amphipod Leptocheirus plumulosus to provide interpretive guidance for sediment toxicity tests with this species. This time-varying, field-based model includes several matrices to reflect seasonal changes in demographics. In this paper, we conduct sensitivity analysis of the model to identify which life history parameters have the greatest potential impact on population growth rate (lambda). Results indicate seasonal variability in the relative demographic importance of vital rates. Over winter, annual population growth is most sensitive to the persistence of juveniles and adults and growth from the juvenile to the adult stage. In spring and fall, changes in fecundity are likely to have large effects on population dynamics. In addition, we demonstrate the applicability of the model by using it to interpret toxicological data from an assessment of sediment contamination in Baltimore Harbor, MD. The model was parameterized with survival data from acute toxicity tests with L. plumulosus to project effects on population growth rate (lambda). Results of these model simulations indicate that relatively small changes in survival can result in large changes in lambda, indicating high risk to benthic populations. Furthermore, population projections mirror observed abundances of L. plumulosus at the test sites. These analyses provide a first indication of the usefulness of the Leptocheirus population model as a tool for exploring ecological effects of sediment contamination. PMID- 11495495 TI - Bioaccumulation and biomarkers in the sea star Asterias rubens (Echinodermata: Asteroidea): a North Sea field study. AB - During two North Sea field trips in March 1995 and September 1996 sea stars, Asterias rubens, were collected at various stations along pollution gradients in order to study the relation between biochemical markers and levels of accumulated contaminants. Biomarkers measured were: cytochrome P450 level, benzo[a]pyrene hydroxylase (BPH) activity, acetylcholinesterase (AChE) activity and DNA integrity. Accumulation levels of heavy metals, polychlorinated biphenyls, organochlorine pesticides and polycyclic aromatic hydrocarbons (PAHs) in the pyloric caeca of sea stars indicate different pollution gradients, influenced by rivers in The Netherlands, UK and Germany. For some contaminants, especially PAHs, relatively high levels were found in the central part of the North Sea (Dogger Bank). On the basis of multivariate statistics, stations near the mouth of the Elbe and the Rhine/Meuse were shown to have different patterns of biomarker responses. Sea stars from stations in coastal zones showed relatively high levels of cytochrome P450 and 'P418', another haemoprotein that is present in most marine invertebrates. The station nearest to the Elbe Estuary showed the lowest BPH and AChE activity. DNA integrity was lower especially in stations near the Dutch coast and in a station near the Tees/Tyne estuaries. Using these biomarkers as early warning signals of exposure and/or adverse effects, this type of monitoring can be used also in the future to study the spatial and temporal trends in the quality of coastal waters. PMID- 11495496 TI - Solid phase synthesis of partially protected tocinoic acid: optimization with respect to resin and protecting groups. AB - A few solid phase and solution approaches of good repute were applied in parallel with the aim to provide optimized routes to Boc- and Fmoc-tocinoic acid (3a and 3c) and the corresponding Tyr(Bu(t)) derivatives (3b and 3d). Boc-tocinoic acid is known to couple with tripeptide amides to give substituted oxytocin precursors in high yields, requiring only Boc-cleavage to furnish the corresponding hormone analogs with minimal loss of material. For comparison, two protected linear hexapeptides (2a and 2b) were prepared on three polystyrene supports, two with acid-labile handles and one a conventional chloromethylated resin, in yields of 62-82 and 58-76%, respectively. The intermediate 2a could be converted to 3a with physical data in agreement with those earlier reported. Similarly, the intermediate 2b was converted to 3b. The highest yields for both 2a and 2b were obtained with a 2-chlorotrityl chloride resin, which in addition provided advantages with respect to overall speed and convenience. Additional syntheses of 3c and 3d on this and of 3c on SASRIN resin, in conjunction with trityl instead of benzyl for side-chain protection of cysteine, were also elaborated. PMID- 11495497 TI - A type-II beta-turn, proline-containing, cyclic pentapeptide as a building block for the construction of models of the cleavage site of pro-oxytocin. AB - Previous studies have indicated that proteolytic activation of pro-hormones and pro-proteins occurs most frequently at the level of basic amino acids arranged in doublets and that the dibasic sites are situated in or next to beta-turns. Investigations utilizing synthetic peptides reproducing the N-terminal processing domain of pro-oxytocin-neurophysin have suggested a close relationship between the secondary structure of the cleavage locus and enzyme recognition, the minimal recognized sequence being the -Pro-Leu-Gly-Gly-Lys-Arg-Ala-Val-Leu- segment of the native precursor. NMR investigations and energy minimization studies have demonstrated that this sequence is organized in two type-II beta-turns involving the -Pro-Leu-Gly-Gly- and -Lys-Arg-Ala-Val- sequences. To further strengthen the above reported hypothesis and to study the role of turn subtypes, a new proline containing cyclic substrate of the processing enzyme, in which the N-terminal side that comes before the Lys-Arg pair is constrained to adopt a type-lI beta turn, has been synthesized. The presence of a type-II beta-turn structure in this cyclic peptide model has been demonstrated by a combined NMR, CD and FT-IR absorption investigation. A preliminary study shows that PC1 is able to recognize and process our constrained substrate. PMID- 11495499 TI - Synthesis and solution characterization of a porphyrin-CCK8 conjugate. AB - In this paper we report the synthesis and a detailed NMR solution characterization of a new CCK8 analogue and its indium(III) complex, PK-CCK8 and In-PK-CCK8. The new compounds contain a porphyrin moiety covalently bound through an amide bond to the side chain of a Lys residue introduced at the N-terminus of CCK8. A molecular dynamics simulation, based on the NMR structure of the complex between CCK8 and the N-terminal extracellular arm of the CCK(A) receptor, is also reported. Both the NMR study and the molecular dynamics simulation indicate that the porphyrin-peptide conjugate might be able to bind to the CCK(A) receptor model. The results of the molecular dynamics calculations show that the conformational features of the CCK8/CCK(A) receptor model complex and of the PK CCK8/CCK(A) receptor-model complex are similar. This evidence supports the view that the introduction of the porphyrin-Lys moiety does not influence the mode of ligand binding to the CCK(A) receptor model. The NMR structure of PK-CCK8 in DMSO consists of a well defined pseudo-helical N-terminal region, while the C-terminal region is flexible. Moreover, the absence of NOE contacts between the porphyrin and the peptide indicates that the macrocyclic ring is directed away from the peptide region involved in the binding with the receptor. PMID- 11495498 TI - Probing the shape of a hydrophobic pocket in the active site of delta-opioid antagonists. AB - The change of selectivity and the induction of antagonism by the insertion of 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic) in the second position of several opioid peptides have led to the interpretation of Tyr-Tic as a specific message domain for delta-opioid antagonists and to the discovery of dipeptides with substantial opioid activity. Selectivity and activity increase enormously when Tyr is substituted by 2',6'-dimethyl tyrosine (Dmt), hinting that the side chain of Dmt fits a hydrophobic cavity of the receptor very tightly and precisely. We have investigated the specificity of this fit by systematic changes of the substituents on the aromatic ring of ryr. Mono- and disubstitutions different from 2',6'- invariably lead to catastrophic decreases of activity. The only substitution compatible with retention of substantial antagonism is 2 methyl. An analysis of the conformational properties of all analogues reveals that substitutions do not affect the global shape of the molecule significantly. Accordingly, it is possible to use the shape of the different side chains to map the hydrophobic cavity of the receptor. The resulting complementary image is funnel shaped. PMID- 11495500 TI - The pharmacokinetics of glycyrrhizic acid evaluated by physiologically based pharmacokinetic modeling. AB - Glycyrrhizic acid is widely applied as a sweetener in food products and chewing tobacco. In addition, it is of clinical interest for possible treatment of chronic hepatitis C. In some highly exposed subjects, side effects such as hypertension and symptoms associated with electrolyte disturbances have been reported. To analyze the relationship between the pharmacokinetics of glycyrrhizic acid in its toxicity, the kinetics of glycyrrhizic acid and its biologically active metabolite glycyrrhetic acid were evaluated. Glycyrrhizic acid is mainly absorbed after presystemic hydrolysis as glycyrrhetic acid. Because glycyrrhetic acid is a 200-1000 times more potent inhibitor of 11-beta hydroxysteroid dehydrogenase compared to glycyrrhizic acid, the kinetics of glycyrrhetic acid are relevant in a toxicological perspective. Once absorbed, glycyrrhetic acid is transported, mainly taken up into the liver by capacity limited carriers, where it is metabolized into glucuronide and sulfate conjugates. These conjugates are transported efficiently into the bile. After outflow of the bile into the duodenum, the conjugates are hydrolyzed to glycyrrhetic acid by commensal bacteria; glycyrrhetic acid is subsequently reabsorbed, causing a pronounced delay in the terminal plasma clearance. Physiologically based pharmacokinetic modeling indicated that, in humans, the transit rate of gastrointestinal contents through the small and large intestines predominantly determines to what extent glycyrrhetic acid conjugates will be reabsorbed. This parameter, which can be estimated noninvasively, may serve as a useful risk estimator for glycyrrhizic-acid-induced adverse effects, because in subjects with prolonged gastrointestinal transit times, glycyrrhetic acid might accumulate after repeated intake. PMID- 11495501 TI - Microspheres as floating drug-delivery systems to increase gastric retention of drugs. AB - Gastric emptying is a complex process, which is highly variable and makes in vivo performance of the drug-delivery systems uncertain. In order to avoid this variability, efforts have been made to increase the retention time of the drug delivery systems for more than 12 h. The floating or hydrodynamically controlled drug-delivery systems are useful in such applications. The present review addresses briefly the physiology of the gastric emptying process with respect to floating drug-delivery systems. In recent years, the multiparticulate drug delivery systems are used in the oral delivery of drugs. One of the approaches toward this goal is to develop the floating microspheres so as to increase the gastric retention time. Such systems have more advantages over the single-unit dosage forms. The development of floating microspheres involves different solvent evaporation techniques to create the hollow inner core. The present review addresses the preparation and characterization of the floating microspheres for the peroral route of administration of the drug. PMID- 11495502 TI - Human alpha-1-glycoprotein and its interactions with drugs. AB - For about half a century, the binding of drugs to plasma albumin, the "silent receptor," has been recognized as one of the major determinants of drug action, distribution, and disposition. In the last decade, the binding of drugs, especially but not exclusively basic entities, to another plasma protein, alpha 1 acid glycoprotein (AAG), has increasingly become important in this regard. The present review points out that hundreds of drugs with diverse structures bind to this glycoprotein. Although plasma concentration of AAG is much lower than that of albumin, AAG can become the major drug binding macromolecule in plasma with significant clinical implications. Also, briefly reviewed are the physiological, pathological, and genetic factors that influence binding, the role of AAG in drug drug interactions, especially the displacement of drugs and endogenous substances from AAG binding sites, and pharmacokinetic and clinical consequences of such interactions. It can be predicted that in the future, rapid automatic methods to measure binding to albumin and/or AAG will routinely be used in drug development and in clinical practice to predict and/or guide therapy. PMID- 11495503 TI - Comparison of image compression viability for lossy and lossless JPEG and Wavelet data reduction in coronary angiography. AB - BACKGROUND: Lossless or lossy compression of coronary angiogram data can reduce the enormous amounts of data generated by coronary angiographic imaging. The recent International Study of Angiographic Data Compression (ISAC) assessed the clinical viability of lossy Joint Photographic Expert Group (JPEG) compression but was unable to resolve two related questions: (A) the performance of lossless modes of compression in coronary angiography and (B) the performance of newer lossy wavelet algorithms. This present study seeks to supply some of this information. METHODS: The performance of several lossless image compression methods was measured in the same set of images as used in the ISAC study. For the assessment of the relative image quality of lossy JPEG and wavelet compression, the observers ranked the perceived image quality of computer-generated coronary angiograms compressed with wavelet compression relative to the same images with JPEG compression. This ranking allowed the matching of compression ratios for wavelet compression with the clinically viable compression ratios for the JPEG method as obtained in the ISAC study. RESULTS: The best lossless compression scheme (LOCO-I) offered a mean compression ratio (CR) of 3.80:1. The quality of images compressed with the lossy wavelet-based method at CR = 10:1 and 20:1 was comparable to JPEG compression at CR = 6:1 and 10:1, respectively. CONCLUSION: The study has shown that lossless compression can exceed the CR of 2:1 usually quoted. For lossy compression, the range of clinically viable compression ratios can probably be extended by 50 to 100% when applying wavelet compression algorithms as compared to JPEG compression. These results can motivate a larger clinical study. PMID- 11495504 TI - Echocardiographic evaluation of right ventricular systolic functions in pure mitral stenosis. AB - Detailed echocardiographic evaluation of right ventricular muscle thickness and systolic functions was performed in twenty two cases of isolated rheumatic mitral stenosis without clinical signs of systemic venous congestion, tricuspid regurgitation or atrial fibrillation. Twenty two age and sex matched normal persons formed the control group. Right ventricular thickness was significantly increased in the patients with mitral stenosis. End-diastolic and end-systolic long axis measurements and areas were significantly increased and fractional shortening of these parameters was significantly reduced in the patient group. Our results show that right ventricular systolic functions are significantly impaired even in absence of clinical signs of systemic venous congestion. This impairment of systolic function did not correlate with pulmonary flow acceleration time. Myocardial involvement in rheumatic process could be one possibility. Systolic movement of the Tricuspid annulus and right ventricular mid cavity short axis dimension were not sensitive in detecting right ventricular systolic dysfunction. PMID- 11495505 TI - The use of echocardiographic colour kinetic wall motion to differentiate broad complex tachycardia. AB - Discrimination between supraventricular tachycardia (SVT) with aberrant conduction from ventricular tachycardia (VT) is vital for the safe and effective management of both conditions. Electrocardiographic algorithms for the differentiation of broad complex tachycardia are complex and difficult to implement in the acute setting, with misdiagnosis occurring in up to 40% of acute presentations. This case study shows the potential for echocardiographic colour kinesis (eck) to support electrocardiographic differentiation. A 74-year old man in sinus rhythm with left bundle branch block (lbbb), a history of myocardial infarction and recurrent sustained VT underwent eck analysis of wall motion propagation during a programmed electrical ventricular stimulation study. Sequential 40 ms time frames of echocardiographic colour coded endocardial wall motion velocity were recorded on video during both induced VT of lbbb configuration and near isochronic atrially paced tachycardia in lbbb. During VT there was initial eck propagation of ventricular septal wall motion from the apex to the atria secondary to electrical depolarisation. During atrially paced tachycardia initial eck motion developed in the interatrial septum and atrial wall followed by propagation in the ventricular endocardial septal wall motion from the atria toward the ventricular apex. This eck technique potentially could be used to support the electrocardiographic diagnosis of a broad complex tachycardia. PMID- 11495506 TI - The clinical significance of reverse redistribution in Tl201 cardiac SPET. AB - In an attempt to evaluate the clinical significance of reverse redistribution in Tl201 cardiac SPET, Tl201 SPET examinations included in the data base of our department have been retrospectively reviewed. PATIENTS: Six hundred ninety eight patients demonstrated the pattern of reverse redistribution. For 237 of these patients' cardiac catheterization data were also available. One hundred and six patients of these had a history of myocardial infarction while the rest of them were referred for angina symptoms. RESULTS: Catheterization showed that the infarction area was perfused by a stenosed but patent vessel in 79 of the 106 patients with a history of myocardial infarction (74.5%). Collateral circulation was evident in 21 of these 106 patients (19.8%). Only six patients (5.6%) showed a totally occluded vessel with no obvious evidence of collateral circulation. Of the 131 patients, with no myocardial infarction history, 14 (10.7%) had normal coronary vessels. The rest of the patients (89.3%) demonstrated stenotic vessels perfusing the same area where reverse redistribution was observed. Sixty-three of these patients have been reexamined after 9-36 months. On 15 of these patients (23.8%) the second scintigram showed ischemia in the same segments where reverse redistribution was initially detected. Another 17 of the above patients had a former scintigraphic examination that showed ischemia on the segments that currently demonstrated reverse redistribution. CONCLUSION: The results of the present retrospective study indicate that, in patients with known coronary disease, the pattern of reverse redistribution frequently indicates the ischemic area. PMID- 11495507 TI - Highly automated segmentation of arterial and venous trees from three-dimensional magnetic resonance angiography (MRA). AB - With the development of new magnetic resonance (MR) contrast agents that have longer persistence in the blood, contrast-enhanced magnetic resonance angiography (MRA) facilitates non-invasive imaging of the cardiovascular system at high resolution in large anatomic volumes. These high resolution 'steady state' images have simultaneous enhancement of both the artery and vein blood pools. Consequently, separation of arteries and veins is an emerging challenge in MRA analysis. Because of the complexity of the vascular structure, manual approaches to cardiovascular tree analysis are impractical. A novel, highly-automated low extremity vessel segmentation and display methodology is reported that consists of five main steps: (1) Binary mask generation, (2) tree-structure generation, (3) optimal vessel path calculation, (4) vessel segment labeling, and (5) conflict resolution. The method's performance was tested in computer phantoms and in in vivo data sets. In the computer-generated phantoms, vessel volume errors ranged from 1.0 to 8.8%. In the in vivo data, the labeling errors ranged from 0.1 to 15.5%. The method provided high quality results in individual data sets and demonstrated segmentation robustness. PMID- 11495508 TI - How well can we separate arteries and veins using automated methods? PMID- 11495509 TI - Variance components of two-dimensional strain parameters in the left-ventricular heart wall obtained by magnetic resonance tagging. AB - This study quantifies variance components of two-dimensional strains in the left ventricular heart wall assessed by magnetic resonance (MR) tagging in 18 healthy xxvolunteers. For a 7-mm tagging grid and homogeneous strain analysis, the intersubject variability and measurement error were estimated, as well as the intra- and interobserver variability. The variance components were calculated for the mean strain of a circumferential sector. The results show that the measurement error was almost equal to the intra-observer variability. With four circumferential sectors of 90 degrees each, approximately 65% of the total variance in epsilonr and epsilonc was due to intersubject variability, the remaining 35% was due to measurement error. With 12 sectors of 30 degrees each, the intersubject variability and measurement error both contributed 50% to the total variance. With 18 sectors of 20 degrees each, only 40% of the total variance was due to intersubject variability. The total variability increased with the number of sectors and therefore the number of sectors used in a study will be a trade-off between segment size (defining spatial resolution) and variability. PMID- 11495510 TI - Imaging of an aberrant right coronary artery with retrospective respiratory-gated 3D-MR angiography. PMID- 11495511 TI - Evaluation of the coronary venous system using electron beam computed tomography. AB - New therapeutic strategies in interventional cardiology and electrophysiology involve the coronary veins. This study examines the potential usefulness of electron beam computed tomography to obtain detailed noninvasive definition of the coronary venous anatomy and of arteriovenous relationships. Electron beam computed tomography allows acquisition and three-dimensional reconstruction of tomographic images of the beating heart with high spatial and temporal resolution. Contrast-enhanced, thin-section electron beam computed tomographic coronary arteriographic images of 34 patients (21 men and 13 women, age 60+/-10 years) were analyzed. The visibility of the coronary veins and their spatial relationship to the coronary arteries were assessed qualitatively on two- and three-dimensional displays. The coronary sinus was visible in 91%, the great cardiac vein in 100%, the middle cardiac vein in 88%, at least one vein overlying the lateral surface of the left ventricle in 97%, the anterior interventricular vein in 97%, and the small cardiac vein in 68%. A left marginal and a left posterior vein were seen in 44%, one of the two in 38%, and neither in 3%. The course of the anterior interventricular vein was parallel to the left anterior coronary artery in 79% and a crossover between the two vessels at an obtuse angle occurred in 12%. Contrast-enhanced electron beam computed tomography imaging of the heart noninvasively provides information on the coronary venous system and arteriovenous relationships that may help guide new interventional procedures. PMID- 11495512 TI - Lupus erythematosus. What's new? PMID- 11495513 TI - Pyogenic granuloma: who are you? PMID- 11495514 TI - Primary cutaneous CD30+ T-cell lymphomas: hypothesis for a highly distinctive clinicobiological behaviour. PMID- 11495515 TI - Dendritic cells in the skin and mucosa: what's new. PMID- 11495516 TI - Itraconazole pulse therapy for the treatment of Candida onychomycosis. AB - In this open label, multicentre trial, 44 patients with clinical and mycological evidence of Candida onychomycosis were treated with itraconazole pulse therapy. Onychomycosis of the toes alone and concomitant disease involving the fingers and toes was treated with three pulses, and onychomycosis of the fingers alone with two pulses. Final evaluation for patients with finger and toe onychomycosis was at 6-9 months and 9-12 months, respectively. There were 29 patients with toe onychomycosis (C. albicans, 27; C. glabrata, one; Candida species, one), 12 patients with finger onychomycosis (C. albicans, two; C. glabrata, one) and three patients had combined toe and finger onychomycosis (C. albicans, two; C. guillermondii, one). In the patients with toe onychomycosis mycological cure was observed in 29 of 32 patients (90.6%). There was complete cure [mycological cure (negative culture and KOH at endpoint evaluation) with clinical cure] or marked improvement (mycological cure with 75% or greater decrease in area of involvement of target nail compared with pretherapy) in 24 of 32 patients (75.0%). All 12 patients with finger onychomycosis alone due to Candida species achieved a mycological cure (100%). In this group of patients complete cure or marked improvement was observed in 11 of 12 patients (91.7%). Itraconazole pulse therapy was well tolerated and no serious adverse events were reported in the patients treated with this triazole. In conclusion, itraconazole pulse therapy is an effective and safe treatment for both finger and toe onychomycosis associated with Candida. PMID- 11495517 TI - Dendritic cells and their role in skin-induced immune responses. AB - This artide gives a brief review on dendritic cells (DC) with regard to their origin, life cyde and functions. The regulation of immune responses by DC functioning as antigen-presenting cells is discussed. Special attention is given to epidermal DC, e.g. Langerhans cells. The perspectives of DC-based therapy are also mentioned. PMID- 11495518 TI - Fluorescence urethroscopy following instillation of 5-aminolevulinic acid: a new procedure for detecting clinical and subclinical HPV lesions of the urethra. AB - OBJECTIVE: To report early clinical experience with intraurethral instillation of 5-aminolevulinic acid (ALA) for the detection of clinical lesions (condyloma acuminata) and subclinical human papillomavirus (HPV) lesions of the urethra, not visible by conventional endoscopy. SUBJECTS AND SETTING: Eighty-four men with clinical diagnosis of condyloma acuminata were examined for urethral HPV lesions at the Department of Urology, Ludwig Maximilian University, Munich, Germany. METHODS: The anogenital areas of the patients were thoroughly examined using a magnifying glass before and after application of 5% acetic acid. Conventional as well as fluorescence urethroscopy were performed 1 h after topical application of 0.1% ALA for 15 min. A sensitive colour charge-coupled device camera for fluorescence video inspection was used with spectral analysis. Biopsies were taken for histological examination and HPV detection by polymerase chain reaction (PCR). RESULTS: Forty-three of 84 men attending our clinic for condyloma acuminata had clinical HPV lesions of the urethra. Condylomas of the proximal urethra were found by conventional endoscopy in eight patients. Fluorescence urethroscopy detected additional subclinical lesions in 13 men. All lesions were HPV infections of the urethra confirmed histologically or by PCR. In nine of these subclinical urethra lesions low-risk HPV types (HPV6, 11, 34) were found. Four lesions were associated with high-risk types (HPV18, 31,52,58). CONCLUSIONS: Fluorescence urethroscopy is a promising diagnostic procedure for detecting subtle clinical and subclinical HPV lesions of the urethra, that are normally not visualized by conventional endoscopy. Generally, urethroscopy is recommended in all cases of externally visible condylomas of the urethra after therapy. PMID- 11495519 TI - The multiple faces of Behcet's disease and its aetiological factors. AB - Behcet's disease (BD) is a chronic, inflammatory multisystemic condition of unknown aetiology. It is clinically characterized by recurrent orogenital ulcerations and skin eruptions; ocular manifestations; arthritis; vasculitis and in some cases neurological and large vessel involvement. Aetiology has not been defined, but genetic, environmental, viral, bacterial and immunological factors have been proposed as causative agents. Treatment includes colchicine, thalidomide, steroids and immunosuppressive agents and it is based on the severity of systemic manifestations, such as central nervous system involvement, arterial aneurysms and thrombosis of the major veins. Mortality is related to major system involvement. In this article the different clinical features, the multiple faces of BD and a list of currently suspected aetiological factors of the disease are discussed, and treatment modalities summarized. PMID- 11495520 TI - The effect of hair loss on quality of life. AB - BACKGROUND: The aim of this study was to quantify the effect of hair loss on quality of life. Patients were recruited from an alopecia support group, and were assessed using the Dermatology Life Quality Index (DLQI) and an adapted version of the DLQI. Financial utility questions, an abbreviated version of the Center for Epidemiologic Studies Depression Scale and open-ended questions were also used. OBSERVATIONS: Seventy (90% response rate) questionnaires were returned. DLQI scores in responders with hair loss (mean score = 8.3, SD = 5.6, range 0-23, n = 70) were similar to those recorded in severe psoriasis. The hair loss continued to have a significant impact on life quality well after the initial event (median duration of hair loss = 138 months +/- 114; range 7-588, n = 70). Forty per cent of patients also felt dissatisfied with the way in which their doctor dealt with them. CONCLUSIONS: This study specifically identifies the feelings of loss of self-confidence, low self-esteem and heightened self consciousness in people affected by hair loss. PMID- 11495521 TI - Allergic reactions to decorative skin paintings. AB - BACKGROUND: Dye mixtures containing henna, P-phenylenediamine (PPD), and indigo are applied to the skin in various parts of the world for religious purposes, social recognition or fun. AIM: To identify eventual allergens responsible for contact sensitization to skin decoration in three young children. Lesions remained strictly limited to the site painted during the summer holiday. None of the children had a previous history of contact dermatitis, but two of them suffered from severe seasonal allergic rhinoconjunctivitis. FINDINGS: Patch testing revealed severe contact hypersensitivity to PPD in all three patients, and one child presented a positive prick test to 1% henna in water. The eruptions resolved following spontaneous elimination of the dyes and topical applications of corticosteroids, but left residual depigmentation. CONCLUSION: PPD is the main allergen identified in allergic reactions to decorative skin paintings. PMID- 11495522 TI - Syringoid eccrine carcinoma. AB - A rare case of a syringoid eccrine carcinoma in a 52-year-old male patient is described. During the first hospitalization, syringoma was diagnosed, both clinically and histologically, keeping in mind the possibility of its malignant alteration. Difficulties in making a diagnosis with histological and immunohistochemical examinations are reported. The possibility of differential diagnosis of primary breast carcinoma and methods of its exclusion are also described. Treatment of the patient was surgical with good results. PMID- 11495523 TI - Eccrine syringofibroadenoma: case report and review of the literature. AB - Eccrine syringofibroadenoma (ESFA) is a rare disorder that shows differentiation toward eccrine sweat apparatus. There is a controversy concerning the pathogenesis and differentiation of this tumour. We report a case of ESFA in a 63 year-old Japanese man. We review the literature presenting a classification, including a newly reported subtype. Clinically and pathogenically, ESFA is probably a group of heterogeneous disorders. PMID- 11495524 TI - Familial erythromelanosis follicularis and chromosomal instability. AB - We report a 17-year-old male patient with erythromelanosis follicularis faciei et colli (EFFC), oral leucokeratosis and diabetes mellitus without islet cell antibody. His sister also had minimal findings of EFFC and minimal follicular papules on her shoulders and extensor surfaces of the arms. The father had only fine follicular papules, but no erythromelanosis. Skin and mucous membrane lesions of the proband were investigated histopathologically. Interestingly, in peripheral lymphocyte cultures of the family members, chromosomal breakage was not observed spontaneously, but it was seen with nitrogen mustard, although this disease may be of autosomal recessive inheritance. Thus, we suggest that EFFC may be a polyaetiological disorder (i.e. familial and environmental) and might be considered one of the chromosomal instability syndromes. PMID- 11495525 TI - Post-apopletic trigeminal trophic syndrome. AB - Trigeminal trophic syndrome is an uncommon clinical entity in which cutaneous trophic ulceration develops with continuous manipulation of trigeminal dermatomes. Patients spontaneously refer picking, rubbing and/or scratching at the affected areas because of hypo-anaesthesia, paraesthesia and/or pain following damage of the sensory trigeminal fibres or nuclei. We herein describe a patient who developed the syndrome as a sequela of brain stem infarction. Diagnosis by scrape cytology in ruling-out basal cell carcinoma and other ulcerative skin diseases is discussed and the importance of neurological examination in disclosing hemi-anaesthesia of trigeminal dermatome(s) is emphasized. PMID- 11495526 TI - Recurrent intravascular papillary endothelial hyperplasia developing from a pyogenic granuloma. AB - Intravascular papillary endothelial hyperplasia (IPEH) is a rare benign vascular tumour. Many histological features are similar to those of low-grade angiosarcoma, a common, but more serious condition. Clinical and histological differentiation is important to avoid overtreatment of this benign condition. We report on a 34-year-old woman who developed recurrent IPEH at the site of a previously excised pyogenic granuloma. PMID- 11495527 TI - Phagocytized apoptotic cells in subcutaneous panniculitis-like T-cell lymphoma. AB - We reported on a case of subcutaneous panniculitis-like T-cell lymphoma (SPTCL) with multiple erythematous nodular lesions on the extremities, trunk and face. Histological examination of an excised lesion revealed a dense infiltrate of markedly atypical T-lymphoid cells expressing the CD8+ phenotype located in the subcutaneous tissue with histiocyte-phagocytizing apoptotic cells. The 'bean-bag' histiocytic cells, the characteristic finding of SPTCL, are considered to be products of haemophagocytosis. In our case the 'bean-bag' cells were produced by phagocytosis of apoptotic bodies, as confirmed by electron microscopy. It is suspected that 'bean-bag' cells are related not to haemophagocytosis but to phagocytosis of apoptotic cells in the CD8+ T-cell type of SPTCL. PMID- 11495528 TI - Multiple linear eccrine spiradenomas with eyelid involvement. AB - Eccrine spiradenoma is an uncommon tumour of the sweat gland and usually occurs as a solitary small painful nodule. Very rarely, it may present as multiple tumours arranged in a linear/zosteriform fashion. We present the case of a 23 year-old female who had multiple spiradenomas arranged linearly from the midline of forehead to the left corner of the mouth and a few lesions also on the margin of the left eyelid. The lesions were probably along the Blaschko's lines. Tumours were excised surgically in stages. Literature search revealed only 13 reported cases of linear/zosteriform/naevoid multiple spiradenomas and only four cases of solitary spiradenoma of the eyelid. The review of reported cases suggests that arrangement of tumours in a linear/zosteriform pattern is an important diagnostic clue for multiple (naevoid) spiradenomas. PMID- 11495529 TI - Linear IgA dermatosis presenting with erythema annulare centrifugum lesions: report of three cases in adults. AB - Linear IgA dermatosis presented with erythema annulare centrifugum lesions in three elderly women. Search for underlying malignancy revealed low-grade B-cell lymphoma in one case. In addition to subepidermal blistering, histology showed a typical mixed infiltrate of granulocytes and eosinophils and, occasionally, papillar microabsesses in one case. In the two other subjects, characteristic subepidermal lining with granulocytes was observed. Immunofluorescence studies confirmed the diagnosis, while autoantibodies characteristic for dermatitis herpetiformis were absent. To our knowledge this is the second report of adult linear IgA dermatosis in association with erythema annulare centrifugum lesions. Our observations concord with several other reports of figurate erythema associated with autoimmune blistering disease and other immune disorders. Common antibody-related immunological mechanisms indicate that the two distinct clinical pictures are probably stages of the same pathogenic entity. PMID- 11495530 TI - Lupus erythematosus/lichen planus overlap syndrome with scarring alopecia. AB - Lupus erythematosus (LE) and lichen planus (LP) may occur as an overlap syndrome (LE/LP). The term comprises a heterogeneous group of patients who have clinical, histological and/or immunopathological characteristics of both diseases at the same time. The disease occurs so infrequently as to limit clinical cases for study. We present a case of scarring alopecia due to LE/LP overlap syndrome. Initial biopsy of the scalp showed scarring alopecia only, with subsequent biopsies showing histological features of LE. Direct immunofluorescence was negative for the LE band, but showed features consistent with LP. We report this case as an uncommon cause of scarring alopecia illustrating the importance of multiple biopsies in the diagnosis of LE/LP overlap syndrome. PMID- 11495531 TI - Anetoderma associated with antiphospholipid antibodies: case report and review of the literature. AB - The association of primary anetoderma (PA) with antiphospholipid antibodies (APAs; with or without criteria of primary antiphospholipid syndrome) has been observed repeatedly and a possible pathogenic significance of this asssociation has been hypothesized. We report the case of a 21-year-old-woman who developed anetodermic lesions on her upper trunk and arms. The presence of APAs was demonstrated, but no diagnostic criteria of systemic lupus erythematosus were present. A review of the literature disclosed 20 previously reported patients presenting this peculiar association. Thrombotic phenomena, represented by spontaneous abortions or venous thrombosis, are a frequent complication in this setting. Taking these observations into account a systematic search for APAs in patients with PA seems advisable. PMID- 11495533 TI - Unusual cutaneous metastases presenting breast cancer: case report. PMID- 11495532 TI - Cutaneous umbilical endosalpingiosis with severe abdominal pain. AB - Cutaneous endosalpingiosis is a very rare entity, and to our knowledge umbilical location has been reported in only one patient who developed salpingectomy shortly after. We report cutaneous umbilical endosalpingiosis associated with severe abdominal pain that disappeared after surgical treatment. PMID- 11495534 TI - Epidermolysis bullosa acquisita responsive to cyclosporin therapy. PMID- 11495535 TI - Multiple epidermoid cysts on photodamaged skin sebaceous gland hyperplasia and senile. PMID- 11495536 TI - Pyoderma gangrenosum after cholecystectomy. PMID- 11495537 TI - Lymphoma en cuirase. PMID- 11495538 TI - Chromoblastomycosis in a patient with chronic obstructive pulmonary disease. PMID- 11495539 TI - Common-sense advice for the treatment of selected nail disorders. PMID- 11495540 TI - Distribution of [3H]GR65630 binding in human brain postmortem. AB - We investigated the distribution of serotonin (5-HT) receptors of type 3 (5-HT3) in human brain areas, by means of the the specific binding of [3H]GR65630. The brains were obtained during autoptic sessions from 6 subjects. Human brain membranes and the binding of [3H]GR65630 were carried out according to standardized methods. The highest density (Bmax +/- SD, fmol/mg protein) of [3H]GR65630 binding sites was found in area postrema (13.1+/-9.7), followed at a statistically lower level, by nucleus tractus solitarius (6.7+/-3.4), nervus vagus (5.5+/-2.1), striatum (4.8+/-2.4) with a progressive decrease in amygdala, olivar nuclei, hippocampus, olfactory bulbus and prefrontal cortex, and then by the other cortical areas and the cerebellum, where no binding was detected. These observations extend previous findings on the distribution of 5-HT3 receptors and confirm interspecies variations that might explain the heterogeneous properties of 5-HT3 receptors in different animals. PMID- 11495541 TI - Effects of linalool on glutamate release and uptake in mouse cortical synaptosomes. AB - Linalool, a monoterpene compound prevalent in essential oil of plant species traditionally used as sedatives, has been characterized as anticonvulsant in several experimental models. Linalool inhibits the binding of [3H]glutamate and [3H]dizocilpine to brain cortical membranes, indicating a participation of the glutamatergic transmission its mechanism of action. In this study, we investigated the effects of linalool on [3H]glutamate release (basal and potassium-stimulated) and [3H]glutamate uptake in mice cortical synaptosomes. Linalool significantly reduced potassium-stimulated glutamate release as well as glutamate uptake, not interfering with basal glutamate release. The data indicates that linalool may interfere with several relevant elements of the glutamatergic transmission, including detriment of the K+-stimulated glutamate release. PMID- 11495542 TI - Transgene delivery with a cationic lipid in the presence of amyloid beta (betaAP) peptide. AB - The ability of a cationic lipid to deliver plasmid DNA (pDNA) in presence of the neurotoxic fragment of amyloid beta-peptide was evaluated. Pre-treatment of cells with betaAP (25-35) peptide resulted in a modest increase in transgene expression. When betaAP (25-35) peptide was mixed with the pDNA/liposome complex and used, the complexes lost their ability to transfect. However, the reverse sequenced betaAP (35-25) peptide demonstrated no significant differences in transgene expression in pre-treated cells, and in cells where betaAP (35-25) peptide was mixed with pDNA/liposome complexes and transfected. The amount of pDNA delivered to the cells was decreased in presence of betaAP (25-35) as measured with flow cytometry using fluorescently labeled liposomes. The decreased endocytosis may be due to their rod-like structure formation as demonstrated by electron microscopy and atomic force microscopy (AFM). These results demonstrate that betaAP (25-35) peptide may interfere with gene delivery with cationic systems. PMID- 11495543 TI - Amyloid beta peptide 1-40 and the function of rat hippocampal hemicholinium-3 sensitive choline carriers: effects of a proteolytic degradation in vitro. AB - Effects of amyloid beta peptide 1-40 (Abeta) and of plant cysteine proteases bromelain and papain on the high-affinity uptake of choline (HACU) and the specific binding of [3H]hemicholinium-3 ([3H]HC-3) have been investigated on hippocampal synaptosomes from young adult male Wistar rats under basal and stimulated conditions (55 mM KCl). Depolarization increased significantly the HACU levels (the changes were predominantly in Vmax) and mildly the [3H]HC-3 binding (the changes especially in K(D)). Nonaggregated Abeta at low nM concentrations suppressed the depolarization effects but was ineffective under basal conditions during a short-term incubation. Higher microM concentrations decreased the HACU and binding under basal conditions in a time-dependent manner. The binding changes were firstly associated with alterations in K(D) and secondarily were accompanied also by a drop in Bmax. The results suggest that Abeta directly influences high-affinity carriers, inhibits their transport activity and enhances their sensitivity to proteoLytic cleavage. Stimulation increases the sensitivity of carriers to the interaction with Abeta. PMID- 11495545 TI - Seizure activity produces differential changes in adenosine A1 receptors within rat hippocampus. AB - Specific ligand binding to rat hippocampal adenosine A1 receptor after administration of the convulsant drug 3-mercaptopropionic acid (MP) was studied by means of a quantitative autoradiographic method. 2-Chloro-N6-[cyclopentyl 2,3,4,5-3H adenosine] ([3H]CCPA), a potent and selective A1 receptor ligand, was selected for binding studies. MP administration (150 mg/kg, i.p.), at seizure, caused significant increases in the following CA1 layers: pyramidal (45%), radiatum (18%) and lacunosum molecular (35%); in CA2 area, a significant decrease in stratum oriens (36%) and an increase in stratum radiatum (14%) and lacunosum molecular (33%) layers was observed. In CA3 area a rise in pyramidal (40%) and radiatum layers (26%), as well as in hillus (97%) was found. At postseizure, changes were restricted to CA1, CA2 and CA3 pyramidal layers and to CA1 lacunosum molecular layer, with increases ranging from 22 to 50%. These results show that [3H]CCPA binding is modified diversely in intrahippocampal layers and areas, thus indicating their dissimilar role in seizure activity. PMID- 11495544 TI - Further characterization of 5-HT1A receptors in the goldfish retina: role of cyclic AMP in the regulation of the in vitro outgrowth of retinal explants. AB - The presence of serotonin 5-HT1A receptors and their physiological role were further characterized in the goldfish retina. The effects of the 5-HT6/7 receptor antagonists pimozide, fluphenazine and amoxapine, the 5-HT1A receptor antagonist WAY-100,135, and the alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline, on the 5-HT1A receptor agonist [3H]8-hydroxy-2-(di-n propylamino)tetralin binding to retinal membranes, were evaluated. In addition, the effects of serotonin, 8-hydroxy-2-(di-n-propylamino)tetralin, WAY-100,135, the adenylate cyclase inhibitors SQ22536 and MDL12330A, and the cyclic AMP analog 8-bromoadenosine-3':5' cyclic monophosphate were also studied on neuritic outgrowth from retinal explants. WAY-100,135 but not 5-HT6/7 receptor antagonists inhibited [3H]8-hydroxy-2-(di-n-propylamino)tetralin binding to retinal membranes N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline decreased [3H]8-hydroxy-2-(di-n propylamino)tetralin binding sites up to 70%, while receptor turnover was similar to that reported in other tissues. Serotonin and 8-hydroxy-2-(di-n propylamino)tetralin stimulated cyclic AMP production, both ex vivo and in vitro, and these increases were related to inhibition of neuritic outgrowth. The inhibitory effect was reduced by SQ22536 and by WAY-100,135, and was mimicked by 8-bromoadenosine-3':5'cyclic monophosphate. This study supports previous findings about the role of serotonin as a regulator of axonal outgrowth during in vitro regeneration of the goldfish retina and demonstrates that this effect is mediated, at least in part, by 5-HT1A receptors through a mechanism which involves an increase of cyclic AMP levels. PMID- 11495546 TI - High intensity light increases olfactory bulb melatonin in Venezuelan equine encephalitis virus infection. AB - In mice infected with the Venezuelan equine encephalomyelitis (VEE) virus and exposed to high intensity light (2500 lux) with a 12 h light: 12 h dark photoperiod, a significant increase in the levels of melatonin in the olfactory bulb was observed. The significance of these findings deserves further studies to understand the mechanisms involved in this effect since the olfactory bulbs have been proposed as first portal for VEE virus entry into the CNS. The increase in melatonin content could represent one of the mechanisms of defense against the viral attack. PMID- 11495547 TI - Alteration in cytoskeletal protein levels in sciatic nerve on post-treatment of diisopropyl phosphorofluoridate (DFP)-treated hen with phenylmethylsulfonyl fluoride. AB - Diisopropyl phosphorofluoridate (DFP) is an organophosphorus ester, and a single dose (1.7 mg/kg, sc.) of this compound produces mild ataxia in hens in 7-14 days and a severe ataxia or paralysis (OPIDN) in three weeks. OPIDN is associated with axonal swelling and their degeneration. We have previously observed alteration in neurofilament (NF) protein levels in the spinal cord of DFP-treated hens. The main objective of this investigation was to study NF protein levels in the sciatic nerves (SN) of hens, in which OPIDN has been potentiated by phenylmethylsulfonyl fluoride (PMSF) post-treatment. PMSF is known to protect DFP treated (1.7 mg/kg) hens from developing OPIDN if injected before, and potentiate OPIDN if injected after the administration of DFP (0.5 mg/kg). The potentiation of OPIDN was accompanied by earlier elevation of NF proteins in the SN particulate fraction. In contrast, SN supernatant fraction showed a transient fall in NF protein levels in potentiation OPIDN. Out of the two other cytoskeletal proteins (i.e., tubulin, tau) studied in this investigation, tubulin also showed earlier elevation in its level in the particulate fraction in potentiated OPIDN. The earlier elevation of NF protein levels in SN particulate fraction in potentiated OPIDN suggested the possible involvement of NFs in delayed neurotoxicity. PMID- 11495548 TI - Glycine, serine, and leucine metabolism in different regions of rat central nervous system. AB - We have investigated the glycine, serine and leucine metabolism in slices of various rat brain regions of 14-day-old or adult rats, using [1-14C]glycine, [2 14C]glycine, L-[3-14C]serine and L-[U-14C]leucine. We showed that the [1 14C]glycine oxidation to CO2 in all regions studied occurs almost exclusively through its cleavage system (GCS) in brains of both 14-day-old and adults rats. In 14-day-old rats, the highest oxidation of [1-14C]glycine was in cerebellum and the lowest in medulla oblongata. In these animals, the L-[U-14C]leucine oxidation was lower than the [1-14C]glycine oxidation, except in medulla oblongata where both oxidations were the same. Serine was the amino acid that showed lowest oxidation to CO2 in all structure studied. In adult rats brains, the highest oxidation of [1-14C]glycine was in cerebral cortex and the lowest in medulla oblongata. We have not seen difference in the lipid synthesis from both glycine labeled, neither in 14-day-old rats nor in adult ones, indicating that the lipids formed from glycine were not neutral. Lipid synthesis from serine was significantly high than lipid synthesis and from all other amino acids studied in all studied structures. Protein synthesis from L-[U-14C]leucine was significantly higher than that from glycine in all regions and ages studied. PMID- 11495549 TI - 2,3-Dimercaptopropanol inhibits Ca2+ transport in microsomes from brain but not from fast-skeletal muscle. AB - Ca2+ is involved in the regulation of a variety of physiological processes, but a persistent increase in free cytosolic Ca2+ concentrations may contribute to cell injury. Dimercaprol (BAL) is a compound used in the treatment of mercury intoxication, but presents low therapeutic efficacy. The molecular mechanism responsible for the BAL toxicity is poorly known. In the present study, the effect of BAL and inorganic and organic mercury on Ca2+ transport by Ca2+-ATPases located in the sarco/endoplasmic reticulum of fast-skeletal muscle and brain was examined. Ca2+ uptake by brain and fast-skeletal muscle microsomes was inhibited in a dose-dependent manner by Hg2+. The calculated IC50 for Ca2+ uptake inhibition by HgCl2 was 1.05+/-0.09 microM (n = 8) for brain and 0.72+/-0.06 microM (n = 9) for muscle. The difference was significant at p < 0.01 (data expressed as mean +/- SD). At a low concentration (1 microM), 2,3-dimer captopropanol had no effect on Ca2+ uptake by brain or muscle vesicles and did not abolish the inhibition caused by Hg2+. A high concentration of BAL (1 mM) nearly abolished the inhibition caused by 1.75 microM HgCl2 or 6 microM CH3HgCl in skeletal muscle. Surprisingly, at intermediate concentrations (40-100 microM) BAL partially inhibited Ca2+ transport in brain but had no effect on muscle. Furthermore, ATP hydrolysis by brain or muscle microsomes was not inhibited by BAL. These results suggest that in brain microsomes BAL affects in a different way Ca2+ transport and ATP hydrolysis. The increase in BAL concentration observed after toxic administration of this compound to experimental animals may contribute to deregulate Ca2+ homoeostasis and, consequently, to the neurotoxicity of BAL. PMID- 11495550 TI - The classic approach to diagnosis of vulvovaginitis: a critical analysis. AB - OBJECTIVE: To correlate the symptoms, signs and clinical diagnosis in women with vaginal discharge, based on the combined weight of the character of the vaginal discharge and bedside tests, with the laboratory diagnosis. METHODS: Women presenting consecutively to the women's health center with vaginal discharge were interviewed and examined for assessment of the quantity and color of the discharge. One drop of the material was then examined for pH and the whiff test was done; a wet mount in saline and in 10% KOH was examined microscopically. The clinical diagnosis was based on the results of these assessments. Gram stain and cultures of the discharge were sent to the microbiology laboratory. RESULTS: One hundred and fifty-three women with vaginal discharge with a clinical diagnosis of vulvovaginitis participated in the study. Fifty-five (35.9%) had normal flora and the other 98 (64.1%) had true infectious vulvovaginitis (kappa agreement = 18%). According to the laboratory, the principal infectious micro-organism causing the vulvovaginitis was Candida species. Candida infection was associated with pH levels of less than 4.5 (p < 0.0001, odds ratio = 4.74, 95% confidence interval: 2.35-9.5, positive predictive value 68.4%). The whiff test was positive in only a small percentage of bacterial vaginosis (BV) (p = not significant (NS)). Clue cells were documented in 53.3% of patients with a laboratory diagnosis of BV (p < 0.02, positive predictive value 26.7%). CONCLUSIONS: The current approach to the diagnosis of vulvovaginitis should be further studied. The classical and time consuming assessments were shown not to be reliable diagnostic measures. PMID- 11495551 TI - Adhesion development and the expression of endothelial nitric oxide synthase. AB - OBJECTIVE: This study was conducted to determine whether nitric oxide (NO), a potent vasodilator and inhibitor of thrombus formation, is involved in the formation and maintenance of adhesions. METHODS: Skin, subcutaneous tissues, peritoneum and adhesions were collected from surgical patients and total RNA was isolated. Quantitative reverse transcription polymerase chain reaction (QRT-PCR) was performed to quantitate endothelial nitric oxide synthase (eNOS) and beta actin mRNA levels. RESULTS: eNOS mRNA levels for skin, subcutaneous tissue, peritoneum and adhesions were < or = 3.12 x 10(-4), < or = 3.12 x 10(-4), 6.24 x 10(-4) and 2.5 x 10(-3) attomoles/microl, respectively. Beta-actin mRNA levels for all tissues were between 1.25 x 10(-1) and 6.25 x 10(-2) attomoles/microl. CONCLUSION: eNOS mRNA can be identified in tissue adhesions, and may therefore play a role in adhesion formation and maintenance. PMID- 11495552 TI - Vaginal mucormycosis: a case report. AB - Although Zygomycetes cause life-threatening, opportunistic infections in immunocompromised hosts, the first case of vaginitis caused by Mucor species in a healthy woman is reported. Mucor vaginitis, which caused mild symptoms only, was refractory to conventional azole therapy and resistant to flucytosine. Cure was achieved with topical amphotericin B. PMID- 11495553 TI - Molecular diagnosis of human papillomavirus: comparison between cervical and vaginal sampling. AB - BACKGROUND: Human papillomavirus (HPV) is the most significant cause of cervical cancer. In view of the number of drawbacks associated with endocervical sampling, the gold standard for HPV detection, this study examined the utility and specificity of vaginal sampling as an alternative for endocervical sampling for the routine detection of HPV. CASE STUDY: The study comprised 51 women who tested positive and 54 women who tested negative for endocervical HPV by polymerase chain reaction (PCR), confirmed by histopathology. At the time of specimen collection, both (speculum-assisted) endocervical and vaginal (no speculum) scrapings were isolated from HPV-positive and negative women, and HPV DNA was assessed by PCR using the MY09/MY11 primer system; HPV type was identified by hybridization of PCR products with type-specific biotinylated DNA probes. Each participant served as her own control. HPV was detected in vaginal and cervical scrapes from all HPV-positive but not HPV-negative women. In HPV-positive women the same HPV type was found in vaginal and endocervical scrapings (positive predictive value = 1.0). CONCLUSION: Correlation between vaginal and endocervical sampling methods was excellent in detecting the presence of HPV DNA and for identifying distinct HPV genotypes. Utilization of vaginal testing for routine HPV detection, and for the long-term follow-up of persistent HPV infection, is therefore recommended. PMID- 11495554 TI - Human papillomavirus typing in HIV-positive women. PMID- 11495555 TI - Immunology update: hepatitis B virus immunization today. PMID- 11495556 TI - Candidiasis during pregnancy may result from isogenic commensal strains. AB - OBJECTIVE: Our laboratory previously demonstrated that asymptomatic vaginal colonization during pregnancy is a factor predisposing patients to subsequent symptomatic vulvovaginal candidiasis. It is unknown whether symptoms result from strain replacement or a change in host relationship to the original colonizing strain. This study was undertaken to determine whether Candida albicans isolates from asymptomatic women could be responsible for subsequent symptomatic vaginitis. METHODS: We retained isolates of C. albicans from women followed longitudinally through pregnancy, and identified six pairs of cultures from women who were colonized without symptoms and who later became symptomatic (average time 14 weeks). We used a random amplification of polymorphic DNA (RAPD) analysis to determine whether isolates from our study patients were genetically similar or dissimilar. RESULTS: Analysis of these pairs of yeast strains by RAPD revealed that five of the six women had symptoms apparently due to the same yeast strain that was found initially as a commensal strain. To increase the power of these observations, we also performed RAPD analysis on six randomly selected yeast strains from other women in this study who had not become symptomatic to determine whether any of these unrelated strains matched strains from those women who became symptomatic. CONCLUSION: Symptomatic yeast vaginitis is usually due to strains of C. albicans already carried in the lower genital tract, underscoring the need to understand regulation of growth and virulence of the organism in vivo. PMID- 11495557 TI - Acyclovir suppression to prevent clinical recurrences at delivery after first episode genital herpes in pregnancy: an open-label trial. AB - OBJECTIVE: To continue evaluation of the use of acyclovir suppression in late pregnancy after first episode genital herpes simplex virus (HSV) infection, using an open-label study design. METHODS: Ninety-six women diagnosed with genital herpes for the first time in the index pregnancy were prescribed suppressive acyclovir 400 mg orally three times daily from 36 weeks until delivery in an open label fashion. Herpes cultures were obtained when patients presented for delivery. Vaginal delivery was permitted if no clinical recurrence was present; otherwise a Cesarean delivery was performed. Neonatal HSV cultures were obtained and infants were followed clinically. Rates of clinical and asymptomatic genital herpes recurrences and Cesarean delivery for genital herpes were measured, and 95% confidence intervals were calculated. RESULTS: In 82 patients (85%) compliant with therapy, only 1% had clinical HSV recurrences at delivery. In an intent to treat analysis of the entire cohort, 4% had clinical recurrences (compared with 18-37% in historical controls). Asymptomatic shedding occurred in 1% of women without lesions at delivery. Two of the four clinical recurrences were HSV culture positive. No significant maternal or fetal side-effects were observed. CONCLUSIONS: In clinical practice the majority of patients are compliant with acyclovir suppression at term. The therapy appears to be effective at reducing clinical recurrences after a first episode of genital herpes complicating a pregnancy. PMID- 11495558 TI - Care of HIV-infected pregnant women in maternal-fetal medicine programs. AB - OBJECTIVE: To survey the evolution over the past decade of attitudes and practices of obstetricians in maternal-fetal medicine fellowship programs regarding the management of human immunodeficiency virus (HIV)-infected pregnant women. METHODS: Directors of all 65 approved maternal-fetal medicine training programs were sent questionnaires, responses to which were to reflect the consensus among members of their faculties. Programs were stratified based upon the number of HIV-infected pregnant patients cared for in the previous year. RESULTS: Responses reflect experience with over 1000 infected pregnant women per year, nearly one-quarter with advanced disease. Combination antiretroviral therapy was prescribed by all respondents, universally in the 2nd and 3rd trimesters. A three-drug regimen (often containing a protease inhibitor) was used more often by those who treated at least 20 HIV-infected pregnant patients per year than by those programs seeing a lower number of patients (80 vs 59%). Despite the known and unknown risks of the use of antiretrovirals during pregnancy, only half of all responding programs report adverse events to the Antiretroviral Pregnancy Registry; reporting was more common among the institutions seeing a higher number of patients (61 vs 45%). Seventy-eight percent of higher volume programs enroll their patients in clinical studies, usually multicenter, versus 35% of lower volume programs. CONCLUSIONS: Care for HIV+ pregnant women has dramatically changed over the past decade. Antiretroviral therapy is now universally prescribed by physicians involved in maternal-fetal medicine training programs. Given limited experience with these agents in the setting of pregnancy, it is essential for maternal-fetal medicine practitioners to actively report on adverse events and participate in clinical trials. PMID- 11495559 TI - Human papillomavirus typing in HIV-positive women. AB - OBJECTIVE: Human papillomavirus (HPV) is the major cause of cervical carcinoma and cervical intraepithelial neoplasia worldwide. Certain HPV types have a strong association with and probably a causative role in the pathogenesis of premalignant cervical lesions. Epidemiologic studies in women infected by the human immunodeficiency virus (HIV) have shown an increased incidence of squamous intraepithelial lesions (SILs), which were predominantly high-grade. Six to 30 per cent of women diagnosed with atypical squamous cells of undetermined significance (ASCUS) on a Papanicolaou (Pap) smear harbor SIL in normal screening populations. This study was undertaken to determine the presence of low-and high risk HPV types in women infected by HIV and to correlate the results to those of the Pap smear. STUDY DESIGN: HPV DNA typing (low- and high-risk) by Digene (Digene Corporation, Gathesburg, MD) hybrid capture methodology was performed on cervical swabs from 209 HIV-positive women. The results of HPV typing were correlated with those of the Pap smear in a retrospective analysis. RESULTS: One hundred and one women (48%) tested positive for HPV subtypes by DNA typing by the hybrid capture method. Of these, 64 patients (63%) had Pap smears which were read as being normal, having benign cellular changes, or having ASCUS (favor reactive process). Of these, 19 patients tested positive for both high-risk and low-risk subtypes, 32 patients tested positive only for high-risk subtypes, and 13 patients tested positive only for low-risk subtypes. CONCLUSION: HPV subtyping identifies a significant group of HIV-positive women who are at risk for developing cervical intraepithelial neoplasia, although they may not show significant abnormalities on their Pap smears. PMID- 11495561 TI - Implant or fixed partial denture? PMID- 11495560 TI - Production of interferons and beta-chemokines by placental trophoblasts of HIV-1 infected women. AB - OBJECTIVE: The mechanism whereby the placental cells of a human immunodeficiency virus (HIV)-1-infected mother protect the fetus from HIV-1 infection is unclear. Interferons (IFNs) inhibit the replication of viruses by acting at various stages of the life cycle and may play a role in protecting against vertical transmission of HIV-1. In addition the beta-chemokines RANTES (regulated on activation T cell expressed and secreted), macrophage inflammatory protein-1-alpha (MIP-1alpha), and MIP-1beta can block HIV-1 entry into cells by preventing the binding of the macrophage-trophic HIV-1 strains to the coreceptor CCR5. In this study the production of IFNs and beta-chemokines by placental trophoblasts of HIV-1 infected women who were HIV-1 non-transmitters was examined. METHODS: Placental trophoblastic cells were isolated from 29 HIV-1-infected and 10 control subjects. Supernatants of trophoblast cultures were tested for the production of IFNs and beta-chemokines by enzyme linked immunosorbent assay (ELISA). Additionally, HIV-1 gag and IFN-beta transcripts were determined by a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) assay. RESULTS: All placental trophoblasts of HIV-1-infected women contained HIV-1-gag transcripts. There were no statistical differences in the median constitutive levels of IFN-alpha and IFN gamma produced by trophoblasts of HIV-1 infected and control subjects. In contrast, trophoblasts of HIV-1-infected women constitutively produced significantly higher levels of IFN-beta protein than trophoblasts of control subjects. Furthermore, the median levels of beta-chemokines produced by trophoblasts of HIV-infected and control women were similar. CONCLUSIONS: Since there was no correlation between the placental HIV load and the production of interferons or beta-chemokines, the role of trophoblast-derived IFNs and beta chemokines in protecting the fetus from infection with HIV-1 is not clear. PMID- 11495562 TI - Guided bone regeneration using bone grafts and collagen membranes. AB - When nonabsorbable membranes are used for guided bone regeneration (GBR), second surgeries are required for membrane retrieval. In addition, these types of membranes show a high incidence of flap sloughing and membrane exposure that often lead to infection and unfavorable results. Absorbable barriers such as collagen membranes were developed to overcome these drawbacks. This article presents the principles and the clinical procedure of using barrier membranes composed of absorbable collagen in GBR aimed at the repair and regeneration of ridge dehiscence defects around implants. The unique properties of collagen membranes that make them ideally suited to GBR procedures are reviewed. In addition, the indications and contraindications for using collagen membranes for GBR procedures are examined. Finally, cases are presented to demonstrate details of surgical principles and techniques. PMID- 11495563 TI - Viability of magnetic denture retainers: a 3-year case report. AB - The purpose of this article is to report on the use of a neodymium-iron-boron magnet enclosed in a laser-welded stainless steel casing as a viable method of retention for removable partial dentures. This mode of retention dispenses with unsightly clasps, at the same time providing direct tooth support via the overdenture abutment. Magnetic attachments are simpler to incorporate into the prosthesis than mechanical attachments, which require springs, coplanar surfaces, or screws. Overdentures of this nature have the added advantage of being more stable and retentive than conventional partial dentures, and they are easily removed and seated without the patient having to grapple with clasps and complex paths of insertion. This particular magnetic attachment exhibited no deterioration after 3 years in function, and the results so far have been promising. PMID- 11495564 TI - The cleft palate patient: a challenge for prosthetic rehabilitation--clinical report. AB - Although cleft palate patients are not regularly seen in general dental practice, their number is not negligible. Prosthodontic treatment of such patients requires good planning that takes into account all remaining teeth and roots, deformation of maxillary segments, residual palatal defects, and the disproportion between the maxillary and mandibular alveolar ridge. With the aim to provide satisfactory function and esthetics and alleviation of the deformities, the authors describe prosthetic therapy of a cleft palate patient using root copings, attachments, telescope and cone crowns, and a metal-base partial prosthesis. The patient's mastication, phonation, and esthetics were improved. Successful results can be best achieved through the judicious use of appropriate treatment modalities tempered by clinical experience and creativity. PMID- 11495565 TI - Amalgam--Resurrection and redemption. Part 1: the clinical and legal mythology of anti-amalgam. AB - Dental amalgam has come under attack for its allegedly poor physical properties and clinical performance in addition to its poor appearance. It has been claimed that the American Dental Association has a hidden agenda to protect amalgam and that other countries have banned its use. A literature search revealed that the vast majority of amalgam restorations do not cause fractured cusps or have recurrent caries. Most amalgam restorations have been shown to last longer than resin composite restorations. In addition, the materials and techniques involved in amalgam restorations have vastly improved in recent years. Like resin composite restorations, amalgam restorations can often be repaired. The American Dental Association has no vested interest in protecting amalgam. The use of amalgam has not been banned in any country in the European Union. According to the latest scientific information available, dental amalgam is a remarkably durable and long-lasting restorative material. Although its appearance is unesthetic, its clinical performance and effectiveness are unsurpassed by those of resin composite. PMID- 11495566 TI - Pulp-dentin biology in restorative dentistry. Part 2: initial reactions to preparation of teeth for restorative procedures. AB - Pulpal complications involving inflammation, degradation, and necrosis are the result of a series of traumatic injuries. The restorative dentist must minimize the trauma to dentin and pulp inflicted during clinical procedures, including that inflicted during tooth preparation. Part 11 of this series discusses the structural and physiologic changes in the pulp-dentin complex that result from crown and cavity preparation and the clinical implication of these changes. PMID- 11495567 TI - Response of three types of cast posts and cores to static loading. AB - OBJECTIVE: An analysis is made of the influence of continuous compression loading on different types of cast posts and cores and its effects on the crowns and teeth. METHOD AND MATERIALS: Thirty extracted maxillary canines were prepared and divided into three groups (n = 10) according to the type of cast posts and cores used for restoration: chrome-nickel alloy, acetalic resin, and IPS-Empress porcelain. Metallic cast crowns were luted onto the cores. The specimens were evaluated with an Instron testing machine at an angulation of 45 degrees and a forward speed of 0.5 mm/min. After testing, the specimens were sectioned along the axis and examined under x40 magnification. RESULTS: Mean load resistance of the restorations until fracture was 2,120 N, 1,491 N, and 2,139 N in the chrome nickel, IPS-Empress ceramic, and acetalic resin groups, respectively. CONCLUSION: Significant differences in resistance were observed between metal and porcelain and between resin and porcelain. Microscopically, the apical portion was found to be most affected by fracture and compromised the teeth subjected to chrome-nickel restorations. The metal cast posts and cores exhibited different angulations, while the ceramic restorations showed cracks. The acetalic resin cast posts and cores presented no fractures. PMID- 11495568 TI - Remineralization across the resin-dentin interface: in vivo evaluation with nanoindentation measurements, EDS, and SEM. AB - OBJECTIVE: The purpose of this study was to evaluate the in vivo remineralization of the possible non-resin infiltrated hybridoid layer between the hybrid layer and the subjacent dentin substrate using nanoindentation, energy dispersive x-ray spectroscopy microanalyses (EDS), and scanning electron microscopy (SEM) technologies. METHOD AND MATERIALS: Twenty Class V cavities were placed in healthy adult monkey teeth. Each cavity was total etched with 37% phosphoric acid for 60 seconds, rinsed, and air dispersed, and SA-Primer was applied to the collagen layer. Cavities were divided into two groups: In group 1, Protect Liner (low-viscosity resin) and Clearfil AP-X (resin composite) were placed per manufacturer's directions, and no bonding agent was placed on the acid-etched interface. In group 2, Clearfil Photobond (bonding agent) was applied, and Protect Liner and Clearfil AP-X were placed as in group 1. Teeth were observed at 7 days (control) and 6 months by nanoindentation, EDS, and SEM. RESULTS: Six month data showed an increased nanohardness in areas 5 pm adjacent to the demineralized or partially demineralized dentin interface. Following treatment with a conventional adhesive system on the acid-etched interface (group 2), there were increased nanohardness and calcium EDS measurements in the substrate just below the resin-dentin impregnated layer. CONCLUSION: Our 6-month in vivo nanoindentation and EDS data demonstrate that the non-resin infiltrated zone becomes remineralized following adhesive resin treatment. PMID- 11495569 TI - Effect of fluoride gels on micromorphology of resin-modified glass-ionomer cements and polyacid-modified resin composites. AB - OBJECTIVE: The purpose of this study was to evaluate the surface micromorphology of resin-modified glass-ionomer cements and polyacid-modified resin composites subjected to a neutral sodium fluoride (NNaF) and an acidulated phosphate fluoride (APF) gel application. METHOD AND MATERIALS: Thirty standardized cylindric specimens were randomly obtained from each of two resin-modified glass ionomer cements-Fuji II LC Improved and Photac-Fil Aplicap-and two polyacid modified resin composites--Dyract and F2000-amounting to 120 samples. After 1 week, the specimens were finished and polished with aluminum oxide disks. Surface treatments with fluoride gels, or distilled water as a control, were performed four times, interspersed with eight pH cycles, simulating high cariogenic challenges. Five calibrated evaluators assessed the surface micromorphology through photomicrographs. RESULTS: The Kruskal-Wallis test showed no significant difference between the control and experimental groups for Fuji II LC Improved and Dyract. Photac-Fil Aplicap showed less micromorphologic change as a result of distilled water application, unlike the NNaF and APF treatments, which revealed no significant difference from each other. For F2000, there was no significant difference between the surfaces treated by NNaF and distilled water; the highest degradation occurred with the APF. CONCLUSION: Both the resin-modified glass ionomer cements and the polyacid-modified resin composites showed erratic behaviors concerning their micromorphology when subjected to fluoride gel application. PMID- 11495570 TI - Remember when? PMID- 11495571 TI - Acromegaly. PMID- 11495572 TI - The fungi in Century 21. The XXI Fungal Genetics Conference, Pacific Grove, California, March 13-18, 2001. PMID- 11495573 TI - The secretion pathway in filamentous fungi: a biotechnological view. AB - The high capacity of the secretion machinery of filamentous fungi has been widely exploited for the production of homologous and heterologous proteins; however, our knowledge of the fungal secretion pathway is still at an early stage. Most of the knowledge comes from models developed in yeast and higher eukaryotes, which have served as reference for the studies on fungal species. In this review we compile the data accumulated in recent years on the molecular basis of fungal secretion, emphasizing the relevance of these data for the biotechnological use of the fungal cell and indicating how this information has been applied in attempts to create improved production strains. We also present recent emerging approaches that promise to provide answers to fundamental questions on the molecular genetics of the fungal secretory pathway. PMID- 11495574 TI - A multiplex PCR test for determination of mating type applied to the plant pathogens Tapesia yallundae and Tapesia acuformis. AB - A multiplex PCR test for determining mating type of the pathogens Tapesia yallundae and Tapesia acuformis is described. The test involves three primers: a "common" primer annealing to DNA sequence conserved in the flanking region of both mating-type idiomorphs and two specific primers annealing to sequence in either the MAT-1 or the MAT-2 idiomorphs. Locating the specific primers in different positions relative to the common primer yielded PCR products of 812 or 418 bp from MAT-1 and MAT-2 isolates, respectively. The test was used successfully to determine the mating type of 118 isolates of T. yallundae and T. acuformis from Europe, North America, and New Zealand. Isolates of both mating types were found on all continents for both species despite the rarely observed occurrence of sexual reproduction of T. acuformis. The multiplex test design should be applicable to other ascomycete species, of use in studies of MAT distribution and facilitating sexual crossing by identifying compatible isolates. PMID- 11495575 TI - Ca(2+) shuttling in vesicles during tip growth in Neurospora crassa. AB - Tip-growing organisms maintain an apparently essential tip-high gradient of cytoplasmic Ca(2+). In the oomycete Saprolegnia ferax, in pollen tubes and root hairs, the gradient is produced by a tip-localized Ca(2+) influx from the external medium. Such a gradient is normally dispensable for Neurospora crassa hyphae, which may maintain their Ca(2+) gradient by some form of internal recycling. We localized Ca(2+) in N. crassa hyphae at the ultrastructural level using two techniques (a) electron spectroscopic imaging of freeze-dried hyphae and (b) pyroantimoniate precipitation. The results of both methods support the presence of Ca(2+) in the wall vesicles and Golgi body equivalents, providing a plausible mechanism for the generation and maintenance of the gradient by Ca(2+) shuttling in vesicles to the apex, without exogenous Ca(2+) influx. Ca(2+) sequestration into the vesicles seems to be dependent on Ca(2+)-ATPases since cyclopiazonic acid, a specific inhibitor of Ca(2+) pumps, eliminated all Ca(2+) deposits from the vesicles of N. crassa. PMID- 11495576 TI - Gene identification in the obligate fungal pathogen Blumeria graminis by expressed sequence tag analysis. AB - Powdery mildew of barley is caused by the obligate fungal pathogen Blumeria graminis f. sp. hordei. Haploid conidia of B. graminis, landing on the barley leaf, germinate to form first a primary germ tube and then an appressorial germ tube. The appressorial germ tube differentiates into a mature appressorium from which direct penetration of host epidermis occurs. Here we present data on 4908 expressed sequence tags obtained from B. graminis conidia. The combined sequences represent 2676 clones describing 1669 individual genes. Comparison with sequences from other pathogenic and nonpathogenic fungi defines hypotheses on the genes required for pathogenicity and growth on the host. The putative roles of some of the identified genes are discussed. PMID- 11495577 TI - The first potent and selective inhibitors of the glycine transporter type 2. PMID- 11495578 TI - Discovery of estrogen sulfotransferase inhibitors from a purine library screen. PMID- 11495580 TI - Rationally based efficacy tuning of selective dopamine d4 receptor ligands leading to the complete antagonist 2-[4-(4-chlorophenyl)piperazin-1 ylmethyl]pyrazolo[1,5-a]pyridine (FAUC 213). AB - Structure dependent efficacy studies in the field of selective D4 ligands led to the 2-aminomethyl substituted azaindole 2 (FAUC 213) that displayed strong D4 binding, high subtype selectivity, and complete antagonist properties in ligand induced mitogenesis experiments. According to our schematic molecular model, the intrinsic activity of the regioisomers investigated is controlled by the ability of the heterocyclic unit to interact with both elements of the D4 binding-site crevice, the aromatic microdomain in TM6, and a serine residue in TM5. PMID- 11495579 TI - Identification of the first trans-(3R,4R)- dimethyl-4-(3-hydroxyphenyl)piperidine derivative to possess highly potent and selective opioid kappa receptor antagonist activity. AB - A structurally novel opioid kappa receptor selective ligand has been identified. This compound, (3R)-7-hydroxy-N-((1S)-1-[[(3R,4R)-4-(3-hydroxyphenyl)-3,4 dimethyl-1-piperidinyl]methyl]-2-methylpropyl)-1,2,3,4-tetrahydro-3 isoquinolinecarboxamide (JDTic, 10) demonstrated high affinity for the kappa receptor in the binding assay (kappa K(i) = 0.3 nM) and highly potent and selective kappa antagonism in the [(35)S]GTP-gamma-S assay using cloned opioid receptors (kappa K(i) = 0.006 nM, mu/kappa ratio = 570, delta/kappa ratio > 16600). PMID- 11495581 TI - Sorbitol dehydrogenase inhibitors (SDIs): a new potent, enantiomeric SDI, 4-[2-1R hydroxy-ethyl)-pyrimidin-4-yl]piperazine-1-sulfonic acid dimethylamide. AB - We report here on our medicinal chemistry and pharmacology efforts to provide a potent sorbitol dehydrogenase inhibitor (SDI) as a tool to probe a recently disclosed hypothesis centered on the role of sorbitol dehydrogenase (SDH) in the second step of the polyol pathway, under conditions of high glucose flux. Starting from a weak literature lead, 2, and through newly developed structure activity relationships, we have designed and executed an unambiguous synthesis of enantiomeric SDI, 6, which is at least 10x more potent than 2. Also, 6 potently inhibits SDH in streptozotocin-diabetic rat sciatic nerve. We have described an expedient synthesis of a key building template, 33, for future research in the SDI area that may facilitate the discovery of even more potent SDIs with longer duration of action in vivo. PMID- 11495582 TI - 3-Phenyl-5-acyloxymethyl-2H,5H-furan-2-ones: synthesis and biological activity of a novel group of potential antifungal drugs. AB - 3-(Substituted phenyl)-5-acyloxymethyl-2H,5H-furan-2-ones related to the natural product (-)incrustoporine were synthesized and their in vitro antifungal activity evaluated. The compounds with halogen substituents on the phenyl ring displayed much higher antifungal effect against Aspergillus fumigatus than selected representatives of azole antifungal drugs. In particular, the activity (1.34 microg/mL) of the most promising derivative, 3-(3,4-dichlorophenyl)-5 pivaloyloxymethyl-2H,5H-furan-2-one, was comparable to that of amphotericin B (0.5 microg/mL). Preliminary evaluation of the toxicity of the compound was carried out as well. Considering the size and properties of these molecules in comparison with those of amphotericin B, further development of this novel group of antifungals may lead to substances with better pharmacological profiles than that of the standard anti-Aspergillus drug. PMID- 11495583 TI - Design, synthesis, and structure-activity relationships of a series of 3-[2-(1 benzylpiperidin-4-yl)ethylamino]pyridazine derivatives as acetylcholinesterase inhibitors. AB - Starting from the 3-[2-(1-benzylpiperidin-4-yl)ethylamino]-6-phenylpyridazine 1, we performed the design, the synthesis, and the structure-activity relationships of a series of pyridazine analogues acting as AChE inhibitors. Structural modifications were achieved on four different parts of compound 1 and led to the following observations: (i) introduction of a lipophilic environment in the C-5 position of the pyridazine ring is favorable for the AChE-inhibitory activity and the AChE/BuChE selectivity; (ii) substitution and various replacements of the C-6 phenyl group are possible and led to equivalent or slightly more active derivatives; (iii) isosteric replacements or modifications of the benzylpiperidine moiety are detrimental to the activity. Among all derivatives prepared, the indenopyridazine derivative 4g was found to be the more potent inhibitor with an IC(50) of 10 nM on electric eel AChE. Compared to compound 1, this represents a 12-fold increase in potency. Moreover, 3-[2-(1-benzylpiperidin 4-yl)ethylamino]-5-methyl-6-phenylpyridazine 4c, which showed an IC(50) of 21 nM, is 100-times more selective for human AChE (human BuChE/AChE ratio of 24) than the reference compound tacrine. PMID- 11495584 TI - 6-Substituted-4-(3-bromophenylamino)quinazolines as putative irreversible inhibitors of the epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor (HER-2) tyrosine kinases with enhanced antitumor activity. AB - A series of new 6-substituted-4-(3-bromophenylamino)quinazoline derivatives that may function as irreversible inhibitors of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor (HER-2) tyrosine kinases have been prepared. These inhibitors have, at the C-6 position, butynamide, crotonamide, and methacrylamide Michael acceptors bearing water-solublilizing substituents. These compounds were prepared by acylation of 6-amino-4-(3 bromophenylamino)quinazoline with unsaturated acid chlorides or mixed anhydrides. We show that attaching a basic functional group onto the Michael acceptor results in greater reactivity, due to intramolecular catalysis of the Michael addition and/or an inductive effect of the protonated basic group. This, along with improved water solubility, results in compounds with enhanced biological properties. We present molecular modeling and experimental evidence that these inhibitors interact covalently with the target enzymes. One compound, 16a, was shown to have excellent oral activity in a human epidermoid carcinoma (A431) xenograft model in nude mice. PMID- 11495585 TI - Fluorosulfonyl- and bis-(beta-chloroethyl)amino-phenylamino functionalized pyrazolo[4,3-e]1,2,4-triazolo[1,5-c]pyrimidine derivatives: irreversible antagonists at the human A3 adenosine receptor and molecular modeling studies. AB - A series of pyrazolotriazolopyrimidines was previously reported to be highly potent and selective human A(3) adenosine receptor antagonists (Baraldi et al. J. Med. Chem. 2000, 43, 4768-4780). A derivative having a methyl group at the N(8) pyrazole combined with a 4-methoxyphenylcarbamoyl moiety at N(5) position, displayed a K(i) value at the hA(3) receptor of 0.2 nM. We now describe chemically reactive derivatives which act as irreversible inhibitors of this receptor. Electrophilic groups, specifically sulfonyl fluoride and nitrogen mustard (bis-(beta-chloroethyl)amino) moieties, have been incorporated at the 4 position of the aryl urea group. Membranes containing the recombinant hA(3) receptor were preincubated with the compounds and washed exhaustively. The loss of ability to bind radioligand following this treatment indicated irreversible binding. The most potent compound in irreversibly binding to the receptor was 14, which contained a sulfonyl fluoride moiety and a propyl group at the N(8) pyrazole nitrogen. The bis-(beta-chloroethyl)amino derivatives displayed a much smaller degree of irreversible binding than the sulfonyl fluoride derivatives. A computer-generated model of the human A(3) receptor was built and analyzed to help interpret these results. The model of the A(3) transmembrane region was derived using primary sequence comparison, secondary structure predictions, and three-dimensional homology building, using the recently published crystal structure of rhodopsin as a template. According to our model, sulfonyl fluoride derivatives could dock within the hypothetical TM binding domain, adopting two different energetically favorable conformations. We have identified two amino acids, Ser247 and Cys251, both in TM6, as potential nucleophilic partners of the irreversible binding to the receptor. PMID- 11495586 TI - Synthesis, mechanism of action, and antiviral activity of a new series of covalent mechanism-based inhibitors of S-adenosyl-L-homocysteine hydrolase. AB - A direct method for the preparation of 5'-S-alkynyl-5'-thioadenosine and 5'-S allenyl-5'-thioadenosine has been developed. Treatment of a protected 5' acetylthio-5'-deoxyadenosine with sodium methoxide and propargyl bromide followed by deprotection gave the 5'-S-propargyl-5'-thioadenosine 4. Under controlled base catalysis with sodium tert-butoxide in tert-butyl alcohol 4 was quantitatively converted into 5'-S-allenyl-5'-thioadenosine 5 or 5'-S-propynyl-5'-thioadenosine 6. Incubation of recombinant human placental AdoHcy hydrolase with 4, 5, or 6 resulted in time- and concentration-dependent inactivation of the enzyme (K(i): 45 +/- 0.5, 16 +/- 1, and 15 +/- 1 microM, respectively). Compound 4 caused complete conversion of the enzyme from its E-NAD(+) to E-NADH form during the inactivation process. This indicates that 4 is a substrate for the 3'-oxidative activity of AdoHcy hydrolase (type I inhibitor). In contrast, the NAD(+)/NADH content of the enzyme was not affected during the inactivation process with 5 and 6, and their mechanism of inactivation was further investigated. Addition of enzyme-sequestered water on the S-allenylthio group of 5 or S-propynylthio group of 6 within the active site should lead to the formation of the corresponding thioester 7. This acylating-intermediate agent could then undergo nucleophilic attack by a protein residue, leading to a type II mechanism-based inactivation. ElectroSpray mass spectra analysis of the inactivated protein by 5 supports this mechanistic proposal. Further studies (MALDI-TOF and ESI/MS(n) experiments) of the trypsin and endo-Lys-C proteolytic cleavage of the fragments of inactivated AdoHcy hydrolase by 5 were carried out for localization of the labeling. The antiviral activity of 4, 5, and 6 against a large variety of viruses was determined. Significant activity (EC(50): 1.9 microM) was noted with 5 against vaccinia virus. PMID- 11495587 TI - Development of serine protease inhibitors displaying a multicentered short (<2.3 A) hydrogen bond binding mode: inhibitors of urokinase-type plasminogen activator and factor Xa. AB - Novel scaffolds that bind to serine proteases through a unique network of short hydrogen bonds to the catalytic Ser195 have been developed. The resulting potent serine protease inhibitors were designed from lead molecule 2-(2-hydroxyphenyl)1H benzoimidazole-5-carboxamidine, 6b, which is known to display several modes of binding. For instance, 6b can recruit zinc and bind in a manner similar to that reported by bis(5-amidino-2-benzimidazolyl)methane (BABIM) (Nature 1998, 391, 608 612).(1) Alternatively, 6b can bind in the absence of zinc through a multicentered network of short (<2.3 A) hydrogen bonds. The lead structure was optimized in the zinc-independent binding mode toward a panel of six human serine proteases to yield optimized inhibitors such as 2-(3-bromo-2-hydroxy-5 methylphenyl)-1H-indole-5-carboxamidine, 22a, and 2-(2-hydroxybiphenyl-3-yl)-1H indole-5-carboxamidine, 22f. Structure-activity relationships determined that, apart from the amidine function, an indole or benzimidazole and an ortho substituted phenol group were also essential components for optimal potency. The affinities (K(i)) of 22a and 22f, for example, bearing these groups ranged from 8 to 600 nM toward a panel of six human serine proteases. High-resolution crystal structures revealed that the binding mode of these molecules in several of the enzymes was identical to that of 6b and involved short (<2.3 A) hydrogen bonds among the inhibitor hydroxyl oxygen, Ser195, and a water molecule trapped in the oxyanion hole. In summation, novel and potent trypsin-like serine protease inhibitors possessing a unique mode of binding have been discovered. PMID- 11495588 TI - Adaptive neuro-fuzzy inference system: an instant and architecture-free predictor for improved QSAR studies. AB - The application of an adaptive neuro-fuzzy inference system (ANFIS) has been developed for obtaining sufficient quantitative structure-activity relationships (QSAR) with high accuracy. To this end, a data set of 68 pyrimidines derivatives as DHFR inhibitors, described first in the excellent independent studies of Hansch et al. (J. Med. Chem. 1982, 25, 777-784 and J. Med. Chem. 1991, 34, 46-54) and later by So and Richards (J. Med. Chem. 1992, 35, 3201-3207), was examined. The ANFIS system, first time applied in the literature to QSAR studies, was trained using a hybrid algorithm consisting of back-propagation and least-squares estimation while the optimum number and shape of membership functions were obtained through the subtractive clustering algorithm. Prior to the development and evaluation of the ANFIS system, geometry optimization of the examined compounds was performed, deriving a series of diverse descriptors from which the best subset was selected by using a hybrid genetic algorithm system. The predictive abilities of the resulting models compared to those produced from classical multivariate regression such as linear and nonlinear (quadratic) partial least squares regression (PLS and QPLS, respectively). The ANFIS method outperformed both the PLS models as well as the published results, leading to substantial gain in both the prediction ability and the computation speed (almost instant training). PMID- 11495589 TI - Human thioredoxin reductase is efficiently inhibited by (2,2':6',2' ' terpyridine)platinum(II) complexes. Possible implications for a novel antitumor strategy. AB - Malignant neoplasms of the brain represent the second leading cause of cancer related mortality in children under the age of 15. The prognosis of patients with glioblastoma multiforme, the most malignant type of gliomas, remains poor offering a median survival time of only 1 year. (2,2':6',2" Terpyridine)platinum(II) complexes are known to possess DNA-intercalating activity and have been shown to be potential chemotherapeutic agents. In the present study we identified the selenoenzyme thioredoxin reductase (TrxR) as a major target of (2,2':6',2"-terpyridine)platinum(II) complexes. New complexes were synthesized in order to optimize this inhibition. The NADPH-reduced enzyme is inhibited almost stoichiometrically by the complexes involving a reversible competitive and an irreversible tight-binding component. For the most potent inhibitor, N,S-bis(2,2':6',2"-terpyridine)platinum(II)-thioacetimine trinitrate, the K(i) for the competitive component of the inhibition is 4 nM and the IC(50) for the tight-binding component is 2 nM after an incubation time of 5 min. The closely related but non-selenium-containing enzyme glutathione reductase is much less inhibited (by a factor of >1000). The platinum complexes were found to strongly inhibit the proliferation of three different glioblastoma cell lines as well as of two different head-and-neck squamous carcinoma cell lines. In a glioblastoma cell culture, less than 10 microM of a platinum(II) compound caused an initial drop of hTrxR activity which was followed by an increase of activity in the surviving cells. A 10 microM inhibitor added every 24 h led to 4% residual hTrxR activity but 100% glutathione reductase activity in the cells surviving for 67 h. The potential of (2,2':6',2"-terpyridine)platinum(II) complexes acting simultaneously at two different intracellular targets-hTrxR and DNA-as antitumor agents is discussed. PMID- 11495590 TI - Toward the design of chemical libraries for mass screening biased against mutagenic compounds. AB - The ability to develop a chemical into a drug depends on multiple factors. Beyond potency and selectivity, ADME/PK and the toxicological profile of the compound play a significant role in its evaluation as a candidate for development. Those factors are being brought into bear earlier in the discovery process and even into the design of libraries for screening. The purpose of our study is the comparative analysis of simple physical characteristics of compounds that have been reported to be mutagens and nonmutagenic ones. The analysis of differences can lead to the development of knowledge-based biases in the libraries designed for massive screening. For each of four Salmonella strains, TA-98, TA-100, TA 1535, and TA-1537, an analysis of the statistical significance of the deviance of the averages for a number of global properties was carried out. The properties studied included parameters, such as topological indices, and bit strings representing the presence or absence of certain chemical moieties. The results suggest that mutagens display a larger number of hydrogen bond acceptor centers for most strains. Moreover, the use of bit strings points to the importance of certain molecular fragments, such a nitro groups, for the outcome of a mutagenicity study. Development of multivariate models based on global molecular properties or bit strings point to a small advantage of the latter for the prediction of mutagenicity. The benefits of the bit strings are in accord with the use of fragment-based approaches for the prediction of carcinogenicity and mutagenicity in methods described in the literature. PMID- 11495591 TI - Prediction of the aroma quality and the threshold values of some pyrazines using artificial neural networks. AB - An artificial neural network is used to predict both the classification of aroma compounds and their flavor impression threshold values for a series of pyrazines. The classification set consists of 98 compounds (32 green, 43 bell-pepper, and 23 nutty smelling pyrazines), and the regression sets consist of 24 green and 37 bell-pepper odorous pyrazines. The best classification of the three aroma impressions (93.7%) is obtained by using a multilayer perceptron network architecture. To predict the threshold values of bell-pepper fragrance, a standard Pearson R correlation coefficient of 0.936 for the training set, 0.912 for the verification set, and 0.926 for the test set is received with two hidden layers consisting of two and one neurons. The network for the threshold prediction of the class of green-smelling pyrazines with one hidden layer containing three neurons turns out to be the best with a standard Pearson R correlation coefficient of 0.859 for the training, 0.918 for the verification, and 0.948 for the test set. These good correlations show that artificial neural networks are versatile tools for the classification of aroma compounds. PMID- 11495592 TI - Methylated analogues of methyl (R)-4-(3,4-dichlorophenylacetyl)- 3-(pyrrolidin-1 ylmethyl)piperazine-1-carboxylate (GR-89,696) as highly potent kappa-receptor agonists: stereoselective synthesis, opioid-receptor affinity, receptor selectivity, and functional studies. AB - Analogues of the kappa-receptor agonist methyl (R)-4-(3,4-dichlorophenylacetyl)-3 (pyrrolidin-1-ylmethyl)piperazine-1-carboxylate (GR-89,696, 6) bearing an additional methyl substituent in the side chain are synthesized and evaluated for their kappa-receptor affinity and selectivity. A key step in the synthesis is the stereoselective reductive amination of the ketones 9, 18, and 19 with pyrrolidine and NaBH(3)CN, which succeeds only in the presence of the Lewis acid Ti(OiPr)(4). Whereas the BOC-substituted ketone 9 affords the unlike and like diastereomers of 10 in a ratio of 70:30, the diastereoselectivity during the reductive amination of the butyl and phenyl substituted ketones 18 and 19 is enhanced to 85:15 (butyl derivative) and >95:<5 (phenyl derivative) in favor of the unlike diastereomers. In receptor binding studies using the radioligand [(3)H]U-69,593 the (S,S) configured methyl carbamate (S,S)-14 reveals the highest kappa-receptor affinity (K(i) = 0.31 nM) within this series, even exceeding the lead kappa-agonist 6 (GR 89,696). A slightly reduced kappa-receptor affinity is observed with the propionamide (S,S)-13 (K(i) = 0.67 nM). The kappa-receptor affinity of piperazines with acyl or alkoxycarbonyl residues at both nitrogen atoms (11, 13, 14) decreases in the order (S,S) > (R,R) > (S,R) > (R,S). The methyl carbamate (S,S)-14 discloses a unique activity profile also binding at mu-receptors in the subnanomolar range (K(i) = 0.36 nM). In a functional assay, i.e., by measuring acetylcholine release in rabbit hippocampus slices, the agonistic effects of the methyl carbamate (S,S)-14 and the propionamide (S,S)-13 are demonstrated. Only weak kappa- and mu-receptor affinities are found with the butyl- and phenyl substituted piperazines 22 and 23. However, considerable sigma(1)-receptor affinity is determined for the enantiomeric, unlike-configured butyl derivatives (R,S)-22 and (S,R)-22 with K(i)-values of 40.2 nM and 81.0 nM, respectively. PMID- 11495593 TI - Synthesis and in vitro and in vivo antimalarial activity of new 4 anilinoquinolines. AB - A new series of 4-anilinoquinolines with two proton-accepting side chains has been synthesized. Antimalarial activity and levels of cytotoxicity upon both MRC 5 cells and macrophages were found to be highly dependent upon the features of these side chains. Several compounds were found to be active in the low nanomolar range, against both chloroquine-sensitive and -resistant strains of Plasmodium falciparum in vitro. From among them, a morpholino derivative cured mice infected by Plasmodium berghei and displayed a lower toxicity than amodiaquine upon mouse macrophages. PMID- 11495594 TI - Selective activation of mitomycin A by thiols to form DNA cross-links and monoadducts: biochemical basis for the modulation of mitomycin cytotoxicity by the quinone redox potential. AB - Mitomycin A (MA) but not mitomycin C (MC) cross-linked linearized (32)P-pBR322 DNA in the presence of dithiothreitol (DTT) or glutathione (GSH), as shown by a sensitive DNA cross-link assay. Incubation of calf-thymus DNA with MA and DTT or mercaptoethanol (MER) resulted in the formation of MA-DNA adducts, which were isolated from nuclease digests of the drug-DNA complexes by HPLC. The adducts were characterized by their UV absorption spectra, electrospray ionization mass spectrometry (ESIMS), and facile conversion from 7-methoxy- to 7-amino substituted mitosene type adducts upon 10% NH(4)OH treatment, which were identical with known adducts of MC. Both DNA interstrand and intrastrand cross link adducts, linking two deoxyguanosine residues at N(2), as well as several deoxyguanosine-N(2) monoadducts of MA, were identified. No DNA adducts were formed with MC under the same conditions. A specificity of DNA cross-link formation for the CpG sequence was observed using 12-mer synthetic oligodeoxyribonucleotides as substrates and as DNA sequence models, in analogy to the known CpG sequence specificity of MC-induced DNA cross-links. MA is known to be more cytotoxic by 2-3 orders of magnitude than MC, and this property correlates with redox potentials of MA (-0.19 V) and MA analogues that are higher than those of MC (-0.40 V) and its analogues. It is suggested that the biochemical basis for the higher cytotoxic potency of MA is MA's propensity to be reductively activated by cellular thiols while MC is resistant to thiol activation. This distinction is probably derived from the large difference between the quinone redox potentials of the two drugs. PMID- 11495595 TI - Synthesis of and a comparative study on the inhibition of muscle and liver glycogen phosphorylases by epimeric pairs of d-gluco- and d-xylopyranosylidene spiro-(thio)hydantoins and N-(d-glucopyranosyl) amides. AB - D-Gluco- and D-xylopyranosylidene-spiro-hydantoins and -thiohydantoins were prepared from the parent sugars in a six-step, highly chemo-, regio-, and stereoselective procedure. In the key step of the syntheses C-(1-bromo-1-deoxy beta-D-glycopyranosyl)formamides were reacted with cyanate ion to give spiro hydantoins with a retained configuration at the anomeric center as the major products. On the other hand, thiocyanate ions gave spiro-thiohydantoins with an inverted anomeric carbon as the only products. On the basis of radical inhibition studies, a mechanistic rationale was proposed to explain this unique stereoselectivity and the formation of C-(1-hydroxy-beta-D glycopyranosyl)formamides as byproducts. Enzyme assays with a and b forms of muscle and liver glycogen phosphorylases showed spiro-hydantoin 12 and spiro thiohydantoin 14 to be the best and equipotent inhibitors with K(i) values in the low micromolar range. The study of epimeric pairs of D-gluco and D-xylo configurated spiro-hydantoins and N-(D-glucopyranosyl)amides corroborated the role of specific hydrogen bridges in binding the inhibitors to the enzyme. PMID- 11495597 TI - Photochemical synthesis of N-arylbenzophenanthridine selective estrogen receptor modulators (serms). AB - Selective estrogen receptor modulators are an emerging class of pharmaceutically important molecules. Many compounds in this class contain an aminoethoxyaryl moiety attached to a polycyclic framework at an asymmetric carbon atom. To assess whether this carbon atom can be replaced by nitrogen, we have employed a Ninomiya enamide photocyclization for the rapid synthesis of a novel N arylbenzophenanthridine framework, 4. Further elaboration of 4 into a new structural class of achiral, nonsteroidal estrogen receptor modulators is described. PMID- 11495596 TI - Synthesis and evaluation of 4-fluoro-8-substituted-2,3,4,5-tetrahydro-1H-2 benzazapines as selective inhibitors of phenylethanolamine N-methyltransferase versus the alpha(2)-adrenoceptor. AB - A small series of 4-fluoro-8-substituted-2,3,4,5-tetrahydro-1H-2-benzazapines (4 fluoro-THBAs; 12-15) were synthesized and evaluated as inhibitors of phenylethanolamine N-methyltransferase (PNMT; EC 2.1.1.28) and as inhibitors of the binding of clonidine at the alpha(2)-adrenoceptor. 4-Fluoro-THBAs 13-15 displayed selectivity ratios (alpha(2) K(i)/PNMT K(i)) greater than 75 and 4 fluoro-8-nitro-THBA (13) was found to be one of the most selective inhibitors of PNMT known, with a selectivity ratio of greater than 900. These compounds are also quite lipophilic and according to previous results from this laboratory should be able to penetrate the blood-brain barrier. These 4-fluoro-THBAs represent important leads in the development of new, more selective, CNS-active inhibitors of PNMT. PMID- 11495599 TI - A piece of my mind: a golden rule: remember the gift. PMID- 11495600 TI - West Nile virus. PMID- 11495601 TI - Chinese and US health care leaders discuss challenges of the 21st century. PMID- 11495602 TI - Hospitals get safety improvement task list. PMID- 11495603 TI - Defense retirees receive better coverage. PMID- 11495608 TI - Statins and fracture risk. PMID- 11495610 TI - Interventions for autism. PMID- 11495612 TI - Depression and hormonal contraception. PMID- 11495614 TI - Factors affecting patients' self-referral to specialists. PMID- 11495616 TI - Time trends in severe head injuries among elderly Finns. PMID- 11495617 TI - Effects of physical activity counseling in primary care: the Activity Counseling Trial: a randomized controlled trial. AB - CONTEXT: Physical activity is important for health, yet few studies have examined the effectiveness of physical activity patient counseling in primary care. OBJECTIVE: To compare the effects of 2 physical activity counseling interventions with current recommended care and with each other in a primary care setting. DESIGN: The Activity Counseling Trial, a randomized controlled trial with recruitment in 1995-1997, with 24 months of follow-up. SETTING: Eleven primary care facilities affiliated with 3 US clinical research centers. PARTICIPANTS: Volunteer sample of 395 female and 479 male inactive primary care patients aged 35 to 75 years without clinical cardiovascular disease. INTERVENTIONS: Participants were randomly assigned to 1 of 3 groups: advice (n = 292), which included physician advice and written educational materials (recommended care); assistance (n = 293), which included all the components received by the advice group plus interactive mail and behavioral counseling at physician visits; or counseling (n = 289), which included the assistance and advice group components plus regular telephone counseling and behavioral classes. MAIN OUTCOME MEASURES: Cardiorespiratory fitness, measured by maximal oxygen uptake (VO(2)max), and self reported total physical activity, measured by a 7-day Physical Activity Recall, compared among the 3 groups and analyzed separately for men and women at 24 months. RESULTS: At 24 months, 91.4% of the sample had completed physical activity and 77.6% had completed cardiorespiratory fitness measurements. For women at 24 months, VO(2)max was significantly higher in the assistance group than in the advice group (mean difference, 80.7 mL/min; 99.2% confidence interval [CI], 8.1-153.2 mL/min) and in the counseling group than in the advice group (mean difference, 73.9 mL/min; 99.2% CI, 0.9-147.0 mL/min), with no difference between the counseling and assistance groups and no significant differences in reported total physical activity. For men, there were no significant between group differences in cardiorespiratory fitness or total physical activity. CONCLUSIONS: Two patient counseling interventions differing in type and number of contacts were equally effective in women in improving cardiorespiratory fitness over 2 years compared with recommended care. In men, neither of the 2 counseling interventions was more effective than recommended care. PMID- 11495618 TI - Risk factors for meningococcal disease in college students. AB - CONTEXT: Elevated rates of meningococcal disease were noted among 18- to 22-year olds in the mid-1990s. However, national data on rates of meningococcal disease in US college students were not collected until 1998. OBJECTIVES: To determine rates of meningococcal disease in US college students and to identify risk factors for meningococcal disease in this population. DESIGN, SETTING, AND PATIENTS: Prospective surveillance study with nested case-control study of US college students with meningococcal infection from September 1, 1998, to August 31, 1999. Fifty state health departments and 231 college health centers participated. MAIN OUTCOME MEASURES: Incidence of and risk factors for meningococcal disease in US college students. RESULTS: Ninety-six cases of meningococcal disease were identified. The incidence rate for undergraduates was 0.7 per 100 000 persons vs 1.4 per 100 000 for the general population of 18- to 23-year-old nonstudents (P<.001). Freshmen living in dormitories had the highest incidence rate at 5.1 per 100 000. Of the 79 case-patients for whom information was available, 54 (68%) had illness due to vaccine-preventable meningococcal serogroups. On multivariable analysis of case-control study data, freshmen who lived in dormitories had an elevated risk of meningococcal disease (matched odds ratio, 3.6; 95% confidence interval, 1.6-8.5; P =.003) compared with other college students. CONCLUSIONS: Freshmen who live in dormitories have an independent, elevated risk of meningococcal disease compared with other college students. Use of the currently available quadrivalent polysaccharide vaccine among college students could substantially decrease their risk of meningococcal disease. PMID- 11495619 TI - Invasive meningococcal disease in adolescents and young adults. AB - CONTEXT: Incidence of invasive meningococcal disease has increased recently in persons aged 15 through 24 years. OBJECTIVE: To characterize meningococcal infection in adolescents and young adults in Maryland during the 1990s. DESIGN AND SETTING: Population-based surveillance study for meningococcal disease from January 1, 1990, through December 31, 1999, in Maryland. PATIENTS: Maryland residents diagnosed as having invasive meningococcal disease. MAIN OUTCOME MEASURE: Invasive meningococcal infection. RESULTS: Of 295 total cases, 71 (24.1%) occurred among persons aged 15 through 24 years. Sixteen (22.5%) of these cases were fatal. The annual incidence rate increased from 0.9 to 2.1 cases per 100 000 among 15 through 24 year olds (P =.01). The proportion of all disease increased from 16.0% to 28.9% (P =.03). The incidence and proportion of cases subsequently decreased to 1.0 and 16.4% in 1998 through 1999, respectively. Infection in 15 through 24 year olds was more likely to be fatal than infection in those younger than age 15 years (22.5% vs 4.6%; P =.001). Infection in 15 through 24 year olds, compared with those aged 25 years or older, was more likely to be associated with male sex (66.2% vs 34.8%; P<.001) and serogroup C infection (46.9% vs 20.2%; P<.001), respectively. Infections were potentially preventable with the licensed meningococcal vaccine in 82.8% of 15 through 24 year olds, 68.1% of those younger than 15 years, and 76.8% of adults aged 25 years or older. CONCLUSIONS: Incidence of meningococcal infection in 15 through 24 year olds in Maryland increased and then declined during the 1990s. Infection in this age group was associated with an unusually high case-fatality ratio, and the vast majority of cases were potentially vaccine preventable. PMID- 11495620 TI - Complications of femoral and subclavian venous catheterization in critically ill patients: a randomized controlled trial. AB - CONTEXT: Whether venous catheterization at the femoral site is associated with an increased risk of complications compared with that at the subclavian site is debated. OBJECTIVE: To compare mechanical, infectious, and thrombotic complications of femoral and subclavian venous catheterization. DESIGN AND SETTING: Concealed, randomized controlled clinical trial conducted between December 1997 and July 2000 at 8 intensive care units (ICUs) in France. PATIENTS: Two hundred eighty-nine adult patients receiving a first central venous catheter. INTERVENTIONS: Patients were randomly assigned to undergo central venous catheterization at the femoral site (n = 145) or subclavian site (n = 144). MAIN OUTCOME MEASURES: Rate and severity of mechanical, infectious, and thrombotic complications, compared by catheterization site in 289, 270, and 223 patients, respectively. RESULTS: Femoral catheterization was associated with a higher incidence rate of overall infectious complications (19.8% vs 4.5%; P<.001; incidence density of 20 vs 3.7 per 1000 catheter-days) and of major infectious complications (clinical sepsis with or without bloodstream infection, 4.4% vs 1.5%; P =.07; incidence density of 4.5 vs 1.2 per 1000 catheter-days), as well as of overall thrombotic complications (21.5% vs 1.9%; P<.001) and complete thrombosis of the vessel (6% vs 0%; P =.01); rates of overall and major mechanical complications were similar between the 2 groups (17.3% vs 18.8 %; P =.74 and 1.4% vs 2.8%; P =.44, respectively). Risk factors for mechanical complications were duration of insertion (odds ratio [OR], 1.05; 95% confidence interval [CI], 1.03-1.08 per additional minute; P<.001); insertion in 2 of the centers (OR, 4.52; 95% CI, 1.81-11.23; P =.001); and insertion during the night (OR, 2.06; 95% CI, 1.04-4.08; P =.03). The only factor associated with infectious complications was femoral catheterization (hazard ratio [HR], 4.83; 95% CI, 1.96 11.93; P<.001); antibiotic administration via the catheter decreased risk of infectious complications (HR, 0.41; 95% CI, 0.18-0.93; P =.03). Femoral catheterization was the only risk factor for thrombotic complications (OR, 14.42; 95% CI, 3.33-62.57; P<.001). CONCLUSION: Femoral venous catheterization is associated with a greater risk of infectious and thrombotic complications than subclavian catheterization in ICU patients. PMID- 11495621 TI - Representation of elderly persons and women in published randomized trials of acute coronary syndromes. AB - CONTEXT: Elderly persons and women were underrepresented in randomized controlled trials (RCTs) prior to 1990. Since then, efforts have been made to correct these biases, but their effect is unclear. OBJECTIVE: To determine whether the percentage of elderly persons and women in published clinical trials of acute coronary syndromes has increased and how this enrollment compared with disease prevalence. DATA SOURCES: The MEDLINE and Cochrane databases were searched for English-language articles from January 1966 to March 2000 regarding myocardial infarction, unstable angina, or acute coronary syndromes. Additional data sources included meta-analyses, review articles, and cardiology textbooks. Estimates of community-based myocardial infarction rates came from the National Registry of Myocardial Infarction and the Worcester Heart Study. STUDY SELECTION: Published RCTs of acute coronary syndrome patients were included and trials enrolling 50 patients or fewer, those without clinical end points, papers published in a language other than English, and unpublished manuscripts were excluded. Of 7645 studies identified, 593 RCTs were selected for review. DATA EXTRACTION: The RCTs were abstracted by 2 of the authors for year of publication, source of support (ie, funding), pharmacotherapy, study phase, number of study sites, trial location, number of patients, mean age of the study population, and any age exclusion criteria for enrollment. DATA SYNTHESIS: The number of published RCTs with explicit age exclusions has declined from 58% during 1966-1990 to 40% during 1991-2000. Trial enrollment of patients aged 75 years or older increased from 2% for studies published during 1966-1990 to 9% during 1991-2000, but remains well below their representation among all patients with myocardial infarction (37%) in the United States. Enrollment of women has risen from 20% for studies published between 1966-1990 to 25% during 1991-2000, but remains well below their proportion of all patients with myocardial infarction (43%) in the United States. CONCLUSIONS: Attempts at making cardiovascular RCTs more inclusive appear to have had limited success; thus, women and elderly persons remain underrepresented in published trial literature relative to their disease prevalence. Because safety and efficacy can vary as a function of sex and age, these enrollment biases undermine efforts to provide evidence-based care to all cardiac patients. PMID- 11495622 TI - Runner's anemia. AB - Macrocytic anemia occurring in patients with fatigue suggests numerous diagnoses, ranging from nutritional deficiencies to a myelodysplastic syndrome. A careful history-taking is critically important for recognition of runner's anemia, which is due to plasma volume expansion, with hemolysis from the pounding of feet on pavement, and hemoglobinuria. Gastrointestinal blood loss may also contribute to anemia in long-distance runners. Early recognition of runner's anemia in patients with a complex presentation of anemia is important in circumventing many diagnostic tests. Runner's anemia should be considered when, amidst a constellation of signs and symptoms, mild anemia is well tolerated by an avid runner. PMID- 11495623 TI - Physical activity counseling in primary care: the challenge of effecting behavioral change. PMID- 11495624 TI - Toward control of meningococcal disease: reducing risk in college students. PMID- 11495629 TI - Evidence for the involvement of Gi2 in activation of extracellular signal regulated kinases in hepatocytes. AB - BACKGROUND: Activation of the extracellular signal-regulated kinases ERK1 and ERK2 in hepatocytes by prostaglandin (PG)F2alpha was recently found to be inhibited by pertussis toxin (PTX) suggesting a role for Gi proteins. RESULTS: Targeting the Gi2alpha expression by a specific ribozyme inhibited the PGF2alpha induced ERK1/2 activation in hepatocytes. On the other hand a non-cleaving form of the Gi2alpha ribozyme did not significantly decrease the ERK1/2 activation. In ribozyme-treated cells the Gi2alpha protein level was reduced, while the Gqalpha level was not affected thus confirming the specificity of the ribozyme. CONCLUSION: The present data suggest an important role of Gi2 in PGF2alpha induced ERK1/2 signaling in hepatocytes. PMID- 11495630 TI - Delta-Notch signaling and lateral inhibition in zebrafish spinal cord development. AB - BACKGROUND: Vertebrate neural development requires precise coordination of cell proliferation and cell specification to guide orderly transition of mitotically active precursor cells into different types of post-mitotic neurons and glia. Lateral inhibition, mediated by the Delta-Notch signaling pathway, may provide a mechanism to regulate proliferation and specification in the vertebrate nervous system. We examined delta and notch gene expression in zebrafish embryos and tested the role of lateral inhibition in spinal cord patterning by ablating cells and genetically disrupting Delta-Notch signaling. RESULTS: Zebrafish embryos express multiple delta and notch genes throughout the developing nervous system. All or most proliferative precursors appeared to express notch genes whereas subsets of precursors and post-mitotic neurons expressed delta genes. When we ablated identified primary motor neurons soon after they were born, they were replaced, indicating that specified neurons laterally inhibit neighboring precursors. Mutation of a delta gene caused precursor cells of the trunk neural tube to cease dividing prematurely and develop as neurons. Additionally, mutant embryos had excess early specified neurons, with fates appropriate for their normal positions within the neural tube, and a concomitant deficit of late specified cells. CONCLUSIONS: Our results are consistent with the idea that zebrafish Delta proteins, expressed by newly specified neurons, promote Notch activity in neighboring precursors. This signaling is required to maintain a proliferative precursor population and generate late-born neurons and glia. Thus, Delta-Notch signaling may diversify vertebrate neural cell fates by coordinating cell cycle control and cell specification. PMID- 11495631 TI - RHD positive haplotypes in D negative Europeans. AB - BACKGROUND: Blood group genotyping is increasingly utilized for prenatal diagnosis and after recent transfusions, but still lacks the specificity of serology. In whites, the presence of antigen D is predicted, if two or more properly selected RHD-specific polymorphism are detected. This prediction must fail, if an antigen D negative RHD positive allele is encountered. Excluding RHDpsi and CdeS frequent only in individuals of African descent, most of these alleles are unknown and the population frequency of any such allele has not been determined. METHODS: We screened 8,442 antigen D negative blood donations by RHD PCR-SSP. RHD PCR positive samples were further characterized by RHD exon specific PCR-SSP or sequencing. The phenotype of the identified alleles was checked and their frequencies in Germans were determined. RESULTS: We detected 50 RHD positive samples. Fifteen samples harbored one of three new Del alleles. Thirty samples were due to 14 different D negative alleles, only 5 of which were previously known. Nine of the 14 alleles may have been generated by gene conversion in cis, for which we proposed a mechanism triggered by hairpin formation of chromosomal DNA. The cumulative population frequency of the 14 D negative alleles was 1:1,500. Five samples represented a D+/- chimera, a weak D and three partial D, which had been missed by routine serology; two recipients transfused with blood of the D+/- chimera donor became anti-D immunized. CONCLUSION: The results of this study allowed to devise an improved RHD genotyping strategy, the false-positive rate of which was lower than 1:10,000. The number of characterized RHD positive antigen D negative and Del alleles was more than doubled and their population frequencies in Europe were defined. PMID- 11495632 TI - The burden of pediatric HIV/AIDS in Constanta, Romania: a cross-sectional study. AB - BACKGROUND: By 1990, 94 percent of the acquired immunodeficiency syndrome (AIDS) cases in Romania were in children less than 13 years of age. The majority of the cases were identified in the city of Constanta. The purpose of this paper was to describe the current burden of pediatric human immunodeficiency virus (HIV) infection in the Constanta county. METHODS: A cross-sectional study was designed to address the primary objective. Between April 1999 and March 2000, all living cases of pediatric HIV infection in the Constanta county were identified from records at the HIV hospital clinic which serves the Constanta county. Standard demographic, social, clinical, treatment and hospitalization data were collected for each study subject. Data were analyzed according to cross-sectional study design methodology. RESULTS: Of the 762 subjects, the majority were seven to 11 years of age, lived with their parents and attended school. Only 70% of the fathers and 13% of the mothers were employed. Horizontal transmission accounted for 90% of the cases. Most of the children had moderate to severe disease as indicated by their AIDS-defining signs; 40% had AIDS. Less than half of the children were receiving antiretroviral therapy (ART). ART and children of mothers with a high school or greater education were independent predictors of long-term non-progression of HIV disease. CONCLUSIONS: This cross-sectional study demonstrated that ten years after the HIV epidemic was identified in Romania, it remains a health and economic burden. The infected children are very ill, but ART is not available for all. The proportion with vertical transmission has increased from an estimated four % to nine %. Our findings support the need to get HIV therapy to economically challenged countries such as Romania. PMID- 11495633 TI - Apolipoprotein E gene polymorphism is not a strong risk factor for diabetic nephropathy and retinopathy in Type I diabetes: case-control study. AB - BACKGROUND: The gene encoding apolipoprotein E (APOE) has been proposed as a candidate gene for vascular complications in Type I diabetes. This study aimed to investigate the influence of three-allelic variations in the APOE gene for the development of diabetic retinopathy and nephropathy. RESULTS: Neither APOE alleles frequencies or APOE genotypes frequencies differed between Type I diabetic groups either with or without nephropathy. Similar results were found for patients with and without diabetic retinopathy. CONCLUSIONS: APOE gene polymorphism does not determine genetic susceptibility for the development of diabetic retinopathy in Type I diabetes patients. Association between APOE gene polymorphism and diabetic nephropathy may be weak or moderate, but not strong. PMID- 11495634 TI - Manipulation of the follicular phase: Uterodomes and pregnancy - is there a correlation? AB - BACKGROUND: Manipulation of the follicular phase uterine epithelium in women undergoing infertility treatment, has not generally shown differing morphological effects on uterine epithelial characteristics using Scanning Electron Microscopy (SEM) and resultant pregnancy rates have remained suboptimal utilising these manipulations. The present study observed manipulation of the proliferative epithelium, with either 7 or 14 days of sequential oestrogen (E) therapy followed by progesterone (P) and assessed the appearance of pinopods (now called uterodomes) for their usefulness as potential implantation markers in seven women who subsequently became pregnant. Three endometrial biopsies per patient were taken during consecutive cycles: day 19 of a natural cycle - (group 1), days 11/12 of a second cycle after 7 days E then P - (group 2), and days 19/22 of a third cycle after 14 days E then P - (group 3). Embryo transfer (ET) was performed in a subsequent long treatment cycle (as per Group 3). RESULTS: Seven pregnancies resulted in seven viable births including one twins and one miscarriage. Analysis of the individual regimes showed 5 days of P treatment to have a higher correlation for uterodomes in all 3 cycles observed individually. It was also observed that all 7 women demonstrated the appearance of uterodomes in at least one of their cycles. CONCLUSIONS: We conclude that manipulation of the follicular phase by shortening the period of E exposure to 7 days, does not compromise uterine epithelial morphology and we add weight to the conclusion that uterodomes indicate a receptive endometrium for implantation. PMID- 11495635 TI - Association of Down's syndrome and water fluoride level: a systematic review of the evidence. AB - BACKGROUND: A review of the safety and efficacy of drinking water fluoridation was commissioned by the UK Department of Health to investigate whether the evidence supported a beneficial effect of water fluoridation and whether there was any evidence of adverse effects. Down's syndrome was one of the adverse effects reported. The aim of this review is to examine the evidence for an association between water fluoride level and Down's syndrome. METHODS: A systematic review of research. Studies were identified through a comprehensive literature search, scanning citations and online requests for papers. Studies in all languages which investigated the incidence of Down's syndrome in areas with different levels of fluoride in their water supplies were included. Study inclusion and quality was assessed independently by 2 reviewers. A qualitative analysis was conducted. RESULTS: Six studies were included. All were ecological in design and scored poorly on the validity assessment. The estimates of the crude relative risk ranged from 0.84 to 3.0. Four studies showed no significant associations between the incidence of Down's syndrome and water fluoride level and two studies by the same author found a significant (p < 0.05) positive association (increased Down's syndrome incidence with increased water fluoride level). Only two of the studies controlled for confounding factors and only one of these presented summary outcome measures. CONCLUSIONS: The evidence of an association between water fluoride level and Down's syndrome incidence is inconclusive. PMID- 11495636 TI - Modulation of L-type Ca2+ current but not activation of Ca2+ release by the gamma1 subunit of the dihydropyridine receptor of skeletal muscle. AB - BACKGROUND: The multisubunit (alpha1S,alpha2-delta, beta1a and gamma1) skeletal muscle dihydropyridine receptor (DHPR) transduces membrane depolarization into release of Ca2+ from the sarcoplasmic reticulum (SR) and also acts as an L-type Ca2+ channel. To more fully investigate the function of the gamma1 subunit in these two processes, we produced mice lacking this subunit by gene targeting. RESULTS: Mice lacking the DHPR gamma1 subunit (gamma1 null) survive to adulthood, are fertile and have no obvious gross phenotypic abnormalities. The gamma1 subunit is expressed at approximately half the normal level in heterozygous mice (gamma1 het). The density of the L-type Ca2+ current in gamma1 null and gamma1 het myotubes was higher than in controls. Inactivation of the Ca2+ current produced by a long depolarization was slower and incomplete in gamma1 null and gamma1 het myotubes, and was shifted to a more positive potential than in controls. However, the half-activation potential of intramembrane charge movements was not shifted, and the maximum density of the total charge was unchanged. Also, no shift was observed in the voltage-dependence of Ca2+ transients. gamma1 null and gamma1 het myotubes had the same peak Ca2+ amplitude vs. voltage relationship as control myotubes. CONCLUSIONS: The L-type Ca2+ channel function, but not the SR Ca2+ release triggering function of the skeletal muscle dihydropyridine receptor, is modulated by the gamma1 subunit. PMID- 11495637 TI - Lowering of lipid composition in aorta of guinea pigs by Curcuma domestica. AB - BACKGROUND: A short-term study was carried out using guinea pigs to determine the effects of Curcuma domestica on lipid composition in the serum and aorta. METHODS: Animals were given food pellets containing 4% (w/w) powdered rhizome of C. domestica in order to determine its effect on cholesterol, triglyceride and phospholipid levels in the aorta and serum. The animals were fed either a cholesterol free diet or a high cholesterol diet (2% cholesterol, w/w, in food pellet) in order to induce hypercholesterolemia. After five weeks of this diet treatment, blood and aorta were taken for biochemical analysis and histological studies. RESULTS: C. domestica in the diet showed no significant effect on the levels of cholesterol, triglyceride and phospholipid in the serum and aorta of the cholesterol free diet animals. However, addition of C. domestica to a high cholesterol diet counteracted increases in the levels of cholesterol, triglyceride and phospholipid in the aorta. Histology studies showed less cholesterol deposits in the aorta of high cholesterol diet animals given C. domestica compared to the high cholesterol diet animals not given C. domestica supplement. C. domestica also had a lowering effect on triglyceride level in the serum of high cholesterol diet animals but showed no effect on serum cholesterol and phospholipid levels. CONCLUSION: This study has shown that dietary intake of C. domestica decreased all lipid composition levels in the aorta and also the serum triglyceride level. In addition, C. domestica also reduced cholesterol deposition in the aorta of high cholesterol diet animals. PMID- 11495638 TI - The impact of indoor residual spraying with malathion on malaria in refugee camps in eastern Sudan. AB - An exploratory trial of the efficacy of indoor spraying with malathion on morbidity and mortality in refugee camps in eastern Sudan was conducted during the rainy season of 1997. The interior walls of houses from a randomly selected group of five camps were sprayed with malathion in mid-September and morbidity and mortality rates in the camps for the months October to December compared with rates in five controls. Pyrethrum spray collection and human landing catches were performed in two collection rounds. An exophagic but endophilic population of Anopheles arabiensis was the most common mosquito collected. The mean human blood index of 242 mosquitoes from eight camps was 0.51. Only two of 1040 mosquitoes examined harboured sporozoites. Blood samples of 83 putative malaria patients were examined for parasites by PCR. Mortality rates in the 3 months following spraying were significantly lower in sprayed camps although differences in clinical malaria incidence between sprayed and non-sprayed camps were not significant. PMID- 11495639 TI - Effect of combined treatment with praziquantel and artemether on Schistosoma japonicum and Schistosoma mansoni in experimentally infected animals. AB - Praziquantel and artemether are safe and efficacious antischistosomal drugs that act against different developmental stages of the parasite: praziquantel against adult worms and artemether against schistosomula. A combined treatment has been suggested as a strategy for transmission control. Recent laboratory experiments with rabbits with a mixed infection of Schistosoma japonicum parasites of different ages confirmed the effectiveness of a combination therapy. In the present work, we assessed the effect of a combined treatment on adult worms of S. japonicum and found significantly higher worm reduction rates than with a single dose of praziquantel. In a next step, we extended the study of the combined treatment to Schistosoma mansoni. A combined treatment with 75 mg/kg praziquantel and 150 mg/kg artemether was administered to hamsters infected with juvenile and adult S. mansoni. The two drugs, administered simultaneously or spaced by 6 h, 1, 3 or 7 days, resulted in significantly higher worm reduction rates than a single treatment with praziquantel. A combination therapy with increased doses of 100 mg/kg praziquantel and 300 mg/kg artemether showed very high worm reduction rates of 90% and above, however, some hamsters died in five different combined treatment experiments, suggesting that these drug concentrations were too high. We conclude that a combined treatment with praziquantel and artemether at the lower doses is safe and more effective than praziquantel alone, which forms a foundation for designing respective clinical trials in humans. PMID- 11495640 TI - Effects of Bay 44-4400, a new cyclodepsipeptide, on developing stages of filariae (Acanthocheilonema viteae, Brugia malayi, Litomosoides sigmodontis) in the rodent Mastomys coucha. AB - Bay 44-4400 was used as a spot on formulation and administered in single doses of 25 and 100 mg/kg to Acanthocheilonema viteae, Brugia malayi, and Litomosoides sigmodontis infected Mastomys coucha on various dates during prepatency, aiming to affect third stage larvae, fourth stage larvae or preadult worms. Microfilaraemia levels were controlled in comparison to untreated controls until necropsies were performed 100 days p.i. (A. viteae, L. sigmodontis) and 150 days p.i. (B. malayi) to determine the numbers of surviving worms and the condition of intrauterine developing stages. A significant proportion (86-100%) of larval and preadult stages of A. viteae were killed by Bay 44-4400 at a dose of 100 mg/kg. A dose of 25 mg/kg had only insignificant effects on the developing parasites, however, it strongly reduced microfilaraemia levels caused by surviving worms in the early phase of patency. Larval and preadult B. malayi and L. sigmodontis were not killed by Bay 44-4400 to a significant degree. Microfilaraemia developing by surviving parasites was generally and significantly reduced throughout the observation period when treatment was performed to affect the preadult parasites. In the other cases variable results were obtained. Intrauterine early embryonic stages were found to be pathologically altered in worms which had been treated at a preadult stage. PMID- 11495641 TI - Long-term sampling of gamma sterilised male Glossina austeni (Diptera: Glossinidae) with sticky panels on Unguja Island (Zanzibar). AB - Daily catches of gamma sterilised male Glossina austeni Newstead with experimental sticky panels were analysed from March 1996 to July 1997. The flies were released weekly by light aircraft over primary and secondary forest ecosystems of Unguja Island, Zanzibar. In the primary forest, the cross-shaped royal blue XT panel (two interlocking panels of each 70 x 60 cm) trapped significantly more flies than the royal blue-white leg panel (panel with a body of 65 x 30 cm and two legs of each 15 x 15 cm) in all months, except in July 1996 and July 1997. In the same habitat, the cross-shaped royal blue-white leg panel (two interlocking leg panels) trapped from 1.7 (not significant) to 3.0 times (highly significant) as many flies as the standard leg panel depending on the season. Significantly more flies were trapped with the cross-shaped XT panel than with the leg panel in the secondary forest compared to the primary forest. Catches of the cross-shaped XT panel on each of a series of days, correlated well with those of the leg panel on the same day, except for panels deployed in the primary forest during the hot-dry and the beginning of the cold-dry season. Catches of the cross-shaped leg panel correlated less well with the catches of the standard leg panel in the primary forest. The data presented indicate that the behavioural responses in time and space of sterile male G. austeni are influenced by the type of trapping device used. PMID- 11495642 TI - Antimalarial activity of azithromycin, artemisinin and dihydroartemisinin in fresh isolates of Plasmodium falciparum in Thailand. AB - Antibiotics with antimalarial activity may offer an interesting alternative for the treatment of multidrug-resistant falciparum malaria. Azithromycin, a relatively recent semisynthetic derivative of erythromycin, was tested for its in vitro activity against fresh isolates of Plasmodium falciparum. As the reportedly slow onset of action of azithromycin suggests its combination with fast-acting substances, such as artemisinin-derivatives, dihydroartemisinin (DHA) was tested parallel as a possible combination partner. The effective concentrations found for azithromycin in this study (EC(50) = 29.3 micromol/l, EC(90) = 77.1 micromol/l blood medium mixture (BMM)) are comparable to those of other antimalarials in the antibiotics class and are considerably higher than those found for mefloquine or quinine. The absence of an activity correlation between azithromycin and chloroquine, quinine and artemisinin emphasises the independence of azithromycin drug response from the sensitivity to these drugs. A weak activity correlation (rho(EC90) = 0.352; p = 0.028), which could point to a potential cross-sensitivity but is probably of little clinical importance, was found with mefloquine above the EC(50) level. Provided that further clinical trials support the combination of these drugs, DHA may offer an interesting combination partner for azithromycin owing to its rapid onset of action and the comparatively low effective concentrations (EC(50) = 1.65 nmol/l, EC(90) = 7.10 nmol/l BMM). This combination may serve as an interesting alternative for tetracycline and doxycycline, which cannot be used in pregnant women and children, and exhibit phototoxicity. Nevertheless, the relatively high cost of this combination, as well as the controversial reports of the clinical efficacy, may limit the usefulness of azithromycin in malaria therapy and require an adjustment of previously used treatment regimens. PMID- 11495643 TI - Intestinal parasites in HIV-2 associated AIDS cases with chronic diarrhoea in Guinea-Bissau. AB - Previous studies from African countries where HIV-1 infection is prevalent have shown that infections with Cryptosporidium parvum, Isospora belli and microsporidia are frequently associated with chronic diarrhoea in AIDS patients. The information about the occurrence of these parasites in HIV-2 associated AIDS cases with chronic diarrhoea is limited. We have performed a study of stool parasites in patients from Guinea-Bissau, the country with the highest prevalence of HIV-2 in the world. Stool specimens from 52 adult patients with chronic diarrhoea of which 37 were HIV-positive and fulfilling the clinical criteria of AIDS (five HIV-1, 28 HIV-2 and four dually infected with HIV-1 and HIV-2) were screened for parasitic infections. Twenty five percent of the HIV-2 positive patients were infected with C. parvum, 11% with I. belli and 11% with microsporidia, all three parasites were seen only in HIV-positive patients. The three patients with microsporidiosis, all HIV-2 infected, are to our knowledge the first cases reported from Guinea-Bissau. Other stool parasites such as Blastocystis hominis, hookworm and Strongyloides stercoralis were observed both among HIV-positive and HIV-negative patients. PMID- 11495644 TI - A simple and quick method for enhanced detection of specific IgE in serum from lymphatic filariasis patients. AB - A new simple and quick technique, using a suspension of protein A agarose beads to absorb IgG4 from sera prior to determination of filarial-specific IgE in ELISA, is presented. The optimal ratio between serum and absorbant was determined by absorbing fixed volumes of sera from individuals from a Wuchereria bancrofti endemic area with different volumes of the protein A agarose bead suspension and testing supernatants for filaria-specific IgG4 and IgE. The effect of absorption on measured IgG4 and IgE intensities in sera from various categories of individuals from the endemic area was thereafter examined. Overall, absorption resulted in a 96.5% decrease in mean ELISA OD values for IgG4 and a 41.6% increase in mean ELISA OD values for IgE. Higher increases in IgE measurements were seen with sera from circulating filarial antigen (CFA) negative individuals (64.7%), microfilaria (mf) negative individuals (56.1%) and individuals with chronic filarial disease (62.7%) than with sera from individuals who were CFA positive (23.4%), mf positive (10.0%), or without chronic disease (36.5%). These differences indicate that the degree to which IgE detection in unabsorbed serum is blocked by IgG4 varies with infection and disease status. Absorption of IgG4 from serum with a protein A agarose bead suspension prior to measurement of specific IgE is a useful alternative to conventional gel column absorption methods, particularly when processing many samples. PMID- 11495646 TI - Occurrence of Leishmania donovani parasitemia in plasma of infected hamsters. AB - Intracardiac transfusion of plasma, mononuclear cell fraction and blood of infected hamster donors induced visceral leishmaniasis in normal hamster receptors. At the moment of transfusion, the donors already showed all the typical signs of the disease: ascites, cachexia, as well as splenomegaly and a high parasite load in the spleen and liver. All transfused hamsters developed typical visceral leishmaniasis between 90 and 120 days, indicating that all blood products were infectious. Transfusion of the mononuclear cell fraction induced the highest values of parasitic load (spleen, 766 Leishman Donovan Units (LDU); liver, 2650 LDU), splenomegaly and hepatomegaly (spleen-liver/body relative weight: 1.130 and 6.870, respectively). Animals that received the plasma fraction also developed visceral leishmaniasis, showing similar parasitic load (spleen, 107 LDU; liver, 220 LDU) and spleen-liver/body relative weight (1.005 and 6.35, respectively) than those transfused with whole blood. The finding of typical Leishmania donovani infection in animals transfused with plasma demonstrates the possibility of the extracellular location of parasites, free in this blood fraction deprived of red and white blood cells. Fluorescence-assisted cell sorter analysis (FACS) of plasma showed the presence of particles corresponding in size to amastigotes, which fluoresced strongly with the serum of a patient with Kala azar (73%), but not with normal serum. PMID- 11495645 TI - Cellular immune responses and cytokine production in BALB/c and C57BL/6 mice during the acute phase of Angiostrongylus costaricensis infection. AB - In our experimental study we were able to show that the contrasting outcome of Angiostrongylus costaricensis infection in C57BL/6 and BALB/c mice, in respect of morbidity and mortality, can be explained by divergent cellular immune responses and a different cytokine pattern in each strain. In BALB/c mice (i.e. those with high mortality), the initial high proliferation of ConA or LPS stimulated spleen cells dropped to very low levels after 2 weeks post-infection (p.i.), whereas in C57BL/6 mice (i.e. those with low mortality), only a minor reduction in lymphoproliferative responses after mitogenic stimulation was observed. The specific proliferation of spleen cells after stimulation with A. costaricensis adult worm antigen remained low in BALB/c mice throughout the experiment, but showed an augmented proliferation in C57BL/6 mice, especially from 2 weeks p.i. onwards. The mitogen-induced production of Th2-type cytokines (IL-4, IL-5, IL-6, IL-10) in spleen cell cultures remained low in BALB/c mice until 4 weeks p.i., but production of Th1-type cytokines (IL-2, IFN-gamma) was highly elevated at 14 and 28 days p.i. In C57BL/6 mice, an upregulated and balanced production of both Th1- and Th2-type cytokines was measured during the course of infection. In summary, a polarization of the immune response towards cellular hyporesponsiveness and a predominantly Th1 cytokine profile was observed in A. costaricensis infected BALB/c mice, which may contribute to pathogenesis and increased morbidity. PMID- 11495647 TI - Giardia trophozoites in dog hookworm, Ancylostoma caninum: accident or hyperparasitism? AB - During the course of transmission electron microscopic studies of adult Ancylostoma caninum removed from a dog, several Giardia trophozoites were found in sections of the buccal cavity, oesophagus and intestine of several hookworms. Although the protozoa appeared viable, this unusual finding probably represents accidental uptake by, rather than an established infection of, the hookworm. It is feasible, however, that the trophozoites might have survived and even multiplied in this aberrant site. PMID- 11495648 TI - Vasopressin into the preoptic area increases grooming behavior in mice. AB - In mice, the neuropeptide arginine-8-vasopressin (AVP) induces excessive grooming, scratching, and hyperactivity when administered intracerebroventricularly. In hamsters, AVP infusion into the medial preoptic area/anterior hypothalamus (MPOA/AH) increases flank marking and flank mark grooming. We measured the behavioral effects of administration of AVP (0, 1, and 10 ng/250 nl) into the preoptic area (POA) of male C57BL/6 mice. Administration of AVP into the POA induced robust effects on grooming, including increased hindleg scratching and face washing. Rearing and olfactory investigation were inhibited by AVP into the POA. These findings indicate that the POA is one site in which AVP induces grooming behavior in mice. PMID- 11495649 TI - The development of olfactory conditioned ejaculatory preferences in the male rat. I. Nature of the unconditioned stimulus. AB - We have demonstrated previously that repeated pairing of a neutral odor with copulation produces a subsequent conditioned ejaculatory preference (CEP) for a female bearing that odor. Here we examine the copulatory components that comprise the unconditioned stimulus (UCS). In Experiment 1, male Long-Evans rats were allowed to copulate with scented females for nine sessions in which they achieved two ejaculations, one ejaculation plus the first intromission following the postejaculatory interval (PEI), one ejaculation without a PEI, or five intromissions without ejaculation. Only the males that achieved two ejaculations or one ejaculation plus the PEI displayed significant CEP. In Experiment 2, males were allowed to remain in the presence of the scented female without access to her after different amounts of copulatory stimulation. Under these conditions, both one and two ejaculations, but not five intromissions, supported the development of CEPs. In Experiment 3, males were allowed to copulate to ejaculation with an unscented female followed by exposure without access to a scented female. This treatment also supported the development of CEP. These results indicate that ejaculation plus a PEI are necessary for the development of CEPs and that the female must be present during the PEI for this to occur. These findings indicate that events during the PEI are the critical components of the UCS for CEP development. PMID- 11495650 TI - The development of olfactory conditioned ejaculatory preferences in the male rat. II. Parametric manipulation of conditioning session number and duration. AB - We have previously demonstrated that repeated pairing of a neutral odor with copulation produces a subsequent conditioned ejaculatory preference (CEP) for females bearing that odor. The present study examines the course of CEP development. In Experiment 1, Long-Evans male rats were allowed access to almond scented, sexually receptive females for either one, five, or nine conditioning sessions that were 30 min in duration. Males given five or nine sessions displayed significant CEPs. In Experiment 2, male rats were given a single conditioning session with multiple almond-scented females until either a duration (60, 120, 180, or 240 min) or copulatory criterion (two, four, or six ejaculatory series) was satisfied. Males that received 120-, 180-, or 240-min sessions or four ejaculations displayed significant CEPs; males that received two or six ejaculations displayed a trend for CEPs. Analysis of effect size estimates revealed that the strongest CEPs were produced by 120 min of copulation or four ejaculations. In Experiment 3, males receiving nine conditioning sessions each 30 min in duration displayed a more enduring CEP than did males receiving a single conditioning session 240 min in duration. These data suggest that early sexual experiences have particularly powerful influences on subsequent sexual preferences and that the development of sexual preferences are influenced by interactions between CS-UCS pairings and motivational variables. PMID- 11495651 TI - Effects of genitofemoral nerve transection on copulatory behavior and fertility in male rats. AB - Selective transection of peripheral nerves that innervate the pelvic region results in the alteration of some aspects of copulatory behavior, including seminal plug weight. Both branches of the genitofemoral nerve primarily innervate the cremaster muscle although the genital branch has a wide distribution in the cremasteric sac. Thermoregulation of spermatogenesis and the mechanical event of ejaculation largely depend on cremaster contractility. In this study, we analyzed the effects of bilateral transection of the genitofemoral nerve on male copulatory behavior and on fertility. Sexually experienced adult male rats were submitted to sexual behavior tests before and after surgical transection of the genitofemoral nerve. Tests were made four times before and on Days 9, 12, 15, 18 and 21 after surgery, and seminal plug weight was also analyzed. In addition, fertility was tested using two different approaches: by allowing the male to ejaculate once in an estrous female or keeping the male with three females during 15 days. Results showed that there were no differences in any of the copulatory parameters, including seminal plug weight. Regarding fertility, both procedures showed that the ability to induce pregnancy was significantly diminished in the neurectomized males tested at different times after surgery. These results support the notion that genitofemoral nerve regulation of fertility via contraction of the male cremaster muscle is an important factor in reproduction in rats. PMID- 11495652 TI - Importance of the environment in conditioned behavior. AB - The purpose of this study was to determine whether an entire experimental situation, and not an individual single stimulus, could be employed to generate a conditioned response. Experiments were conducted on four adult cats. Each cat was conditioned in two different experimental environments (Situation I and Situation II), which consisted of two compartments that differed with respect to color, shape and illumination. Experiments were carried out separately on each cat; experimental sessions, which lasted 30 min, were conducted two or three times a week. Two cats (Nos. 1 and 2) were first trained during 15 sessions in Situation I, and then during 15 subsequent sessions in Situation II. During each session in Situation I, electrical stimulation, which was applied to the basal forebrain area (BFA), evoked a slow-wave EEG pattern; in addition, the animals would close their eyes and lie down (i.e., they exhibited typical presleep behavior). After three to five sessions, this behavior began to appear as soon as the cats were introduced into the experimental compartment, even before stimulation was applied. In a further series of sessions, the cats were placed for 15 sessions in Situation II, wherein stimulation was applied to the lateral hypothalamus (LH). Stimulation of the LH evoked a desynchronized EEG pattern that was accompanied by excitatory behavior. In the other two cats (Nos. 3 and 4), the animals were first trained for 15 sessions in Situation II; subsequently, they were trained for 15 sessions in Situation I. Finally, a test of cross-stimulation was performed. Stimulation of the BFA (which was previously used in Situation I) was applied, one time only, to cats 1 and 2 in Situation II; stimulation of the LH (which was previously used in Situation II) was applied, once only, to cats 3 and 4 in Situation I. In both cases, the animals did not exhibit any of the previously observed behavioral reactions or EEG patterns of activity. The preceding results confirm our hypothesis that, in each situation, a conditioned reaction was established in response to the totality of the experimental environment. PMID- 11495653 TI - Intrameal hepatic-portal infusion of glucose reduces spontaneous meal size in rats. AB - To test whether glucose (GLC) or insulin (INS) acutely reduces spontaneous meal size, we tested the effects of remotely controlled, intrameal hepatic-portal vein infusions of GLC or INS on rats' spontaneous feeding patterns. Experiment 1 included four blocks of three test infusions and one control infusion. The test infusions in each block were 0.25, 0.5, 1.0 or 1.5 mmol GLC; 2, 4, 6, or 8 mU INS; or the four combinations with dose ratios of 1 mmol GLC/8 mU INS, respectively. Control infusions and the INS vehicle were saline infusions that were equiosmotic to the GLC infusion used in that block. Infusions (0.1 ml x 5 min) were done during the first spontaneous dark-phase meal. None of the test infusions affected meal size, meal duration or the duration of the subsequent intermeal interval. In Experiment 2, a similar design was used to test infusions of 1 mmol GLC, 2 mU INS and GLC/INS. Both GLC alone and GLC/INS reduced the size and duration of the first spontaneous dark-phase meal. The subsequent intermeal interval was unaffected, but GLC alone also increased the satiety ratio (min/g) of the meal. The size and duration of the second dark-phase meal were unaffected. INS alone did not affect any meal parameters. In Experiment 3, infusions of 1 mmol GLC and 2 mU INS were repeated during each of the first three meals of the dark phase. These infusions reduced the size and duration of each meal, as well as 6-h cumulative food intake, but did not affect any other meal parameter. These experiments demonstrate for the first time that intrameal hepatic-portal infusions of GLC or of GLC and INS is sufficient to acutely and selectively reduce spontaneous meal size in the rat. The findings are consistent with the idea that meal-contingent changes in hepatic-portal GLC concentration contribute to satiation. PMID- 11495654 TI - Chemosensory responses to sugar and fat by the omnivorous lizard Gallotia caesaris: with behavioral evidence suggesting a role for gustation. AB - Many lizards can identify food using only chemical cues, as indicated by tongue flicking for chemical sampling and biting, but the effectiveness of the chemical components of food are unknown, as is the relationship between response strength and concentration. We investigated responses by the omnivorous lizard Gallotia caesaris to representatives of two major categories of organic food chemicals, lipids and carbohydrates. The stimuli, pork fat and sucrose solutions of varying concentration, were presented to lizards on cotton swabs and their lingual and biting behaviors were observed during 60-s tests. In the first experiment, fat elicited more tongue-flicks and bites than saturated sucrose or water (odorless control), biting being limited to the fat condition. Lizards licked at high rates, but exclusively in response to sucrose. A lick was a lingual protrusion in which the dorsal surface of the tongue contacted the swab, in contrast to the anteroventral contact made during tongue-flicks. In a second experiment, the number of licks, but not the number of tongue-flicks, increased with the concentration of sucrose. The results indicate that lipids contribute to prey chemical discrimination and are adequate to release some attacks, but are not as effective as releasers of attack as mixtures of prey chemicals obtained from prey surfaces. The findings with respect to licking are novel, and suggest that licking may be a response to gustatory stimulation by sugar, in contrast to previously observed prey chemical discriminations shown to require vomerolfaction. PMID- 11495655 TI - Activity of body energy regulatory pathways in inflammation-induced anorexia. AB - Previous research has shown that reductions in body weight prior to induction of acute inflammation can attenuate inflammation-induced anorexia in male rats. In the current study, potential mechanisms responsible for this observation were examined. Specifically, the effect of a 12% prior reduction in body weight on serum leptin, insulin, and corticosterone; levels of interleukin-1 (IL-1), interleukin-6 (IL-6) in the serum, liver, and spleen; neuropeptide Y (NPY) and POMC mRNA levels in the arcuate nucleus (ARC) of the hypothalamus were examined 8 h after induction of acute inflammation. Rats with prior weight reduction had significantly lower serum leptin levels and gene expression of POMC in the ARC than normal-weight rats. In contrast, prior weight reduction altered neither NPY mRNA in the ARC, nor IL-1alpha, IL-1beta, and IL-6 levels in the serum, liver, and spleen. These results suggest that the attenuation of inflammation-induced anorexia by prior weight reduction is not due to altered cytokine activity, but rather to changes in energy regulatory systems that moderate the anorexic actions of IL-1beta and IL-6. One potential change may be reduced activity of the CNS melanocortin system induced by decreased circulating leptin. PMID- 11495656 TI - Restraint stress is associated with changes in glucocorticoid immunoregulation. AB - Psychological stress has been associated with activation of the hypothalamic pituitary-adrenal (HPA) axis and impaired cell-mediated immune (CMI) responses. There is also evidence suggesting that intermittent chronic stress differentially alters CMI across different immune compartments, but the mechanisms underlying this phenomenon have not been explored in detail. In the present study, we investigated (i) acute and chronic restraint stress effects in Sprague-Dawley rats on both peripheral blood lymphocyte (PBL) and splenocyte mitogen-induced proliferation and (ii) also determined whether differential stress effects within these immune compartments might reflect alterations in lymphocyte sensitivity to glucocorticoids. It was found that while acute stress exposure significantly raised plasma corticosterone levels (1048% vs. controls, P<.001), this response was attenuated in the animals previously exposed to chronic intermittent stress ( 79.66% vs. acute; P<.001). Acute stress increased phytohemagglutinin (PHA) induced lymphocyte proliferation in the spleen (69.04%, P=.01) and suppressed PBL proliferation (-45.52%, P<.001). Neither of these changes were observed following chronic stress. We also demonstrated that reexposure to the stressor rapidly increased splenocyte sensitivity to in vitro dexamethasone (P<.05) and corticosterone (P<.05) in chronically stressed rats. Our data (1) confirm that acute stress is associated with compartment-specific changes in CMI function, (2) indicate that chronic stress is associated with habituated endocrine and immune responses and (3) that stressor exposure rapidly alters splenocyte sensitivity to glucocorticoids and we suggest that the latter may contribute to differential stress effects across immune compartments. PMID- 11495657 TI - The actual, but not labelled, fat content of a soup preload alters short-term appetite in healthy men. AB - The effects of the actual and labelled fat content of a soup preload on appetite at a test meal 30 min later were assessed in 16 healthy men. Each participant ate lunch on four occasions, combining two levels of fat energy (Low, 265 kJ or High, 1510 kJ) and two types of label (Low-fat or High-fat), presented as fictitious soup brand names. Preliminary work established that the Low-fat labels produced an expectation of reduced fat content and lower anticipated hedonic ratings, whereas the High-fat labels generated expectations of a high-fat content and above average hedonic ratings. These expectancies were confirmed in the main experiment, with the soups labelled as high fat rated as both more pleasant and creamy than those labelled low-fat, independent of actual fat content. However, intake at the test meal was unaffected by the preload label, but instead reflected the actual fat (hence, energy) content of the soup, with significantly lower food intake after the high-fat soup regardless of the food label. Rated hunger was lower, and fullness higher, at the start of the meal after the high fat preloads regardless of how they were labelled, while the pattern of appetite change during the test meal was unaffected by preload. These results suggest that realistic food labels can modify the immediate experience of a consumed food, but do not alter appetite 30 min later in healthy men. PMID- 11495658 TI - Adaptation to social isolation. Acute and long-term stress responses of growing gilts with different coping characteristics. AB - The present experiment studied the acute and long-term stress responses of reactive and proactive prepubertal gilts to social isolation. Gilts with either reactive or proactive features were identified according to behavioral resistance in a backtest at a young age (2-4 days), respectively being low (LR) and high resistant (HR) in this test. At 7 weeks of age, 12 gilts of each type were socially isolated. Initially, isolation was stressful for both types of gilts, as shown by increased cortisol concentrations and decreased body temperatures. Moreover, both types reacted with increases in exploration and vocalizations. Stress responses to isolation, however, differed in magnitude and/or duration between LR and HR gilts, which was in line with expected reaction patterns on the basis of preferred ways of coping. The cortisol response to isolation was higher in LR gilts, and they generally showed more explorative behavior. HR gilts seemed to be more engaged in walking/running behavior in the first hour after isolation, they generally vocalized more and their noradrenaline excretion in urine was higher at 3 weeks after the start of isolation. Several responses to isolation in the longer term pointed to a prolonged higher general state of stress of HR gilts. Body temperature in HR gilts, for instance, did not recover during 3 weeks of isolation, but values returned to "normal" within 1 day in LR gilts. At 1 week of isolation, relatively high parasympathetic responsivity to novelty was observed in HR gilts, probably due to stress-related high sympathetic reactivity. A shift in percentages of leucocyte subsets, typically occurring under conditions of stress, only developed in HR gilts during isolation. Finally, gastric ulceration was found in one HR gilt, but did not occur in LR gilts. To conclude, LR and HR gilts differed in their strategies to adapt to social isolation, and especially for HR gilts, this procedure seemed to become a chronic stressor. PMID- 11495659 TI - Effects of mild food deprivation on the estrous cycle of rats. AB - It has long been known that severe food deprivation disrupts the estrous cycle. One of the main problems with behavioral tasks that use food for reinforcement is the requirement that the animal be food deprived. This manipulation could be problematic in studies using female animals, since it may interfere with the estrous cycle of the animals. The purpose of the present study was to investigate: (1) the effect of mild food deprivation on four different strains of rats, (2) factors in the food deprivation procedure that could affect the estrous cycle, and (3) the possible effect of enriched diets during food deprivation on the estrous cycle. A comparison of the estrous cycle in four different rat strains revealed differences in the reliability of the estrous cycle even before the onset of food deprivation. Fischer, Long-Evans, and Sprague-Dawley rats all showed reliable cycle patterns. This was not the case for Brown Norway rats. During food deprivation, the cycle of the Fischer rats was disrupted, whereas the Long-Evans and Sprague-Dawley animals continued to cycle. Both the rate of weight loss and the percent of ad libitum body weight were related to cessation of the estrous cycle. However, enriching an animal's diet with sugar or oil additives delayed the disruption of the estrous cycle. Additionally, animals resumed cycling when returned to ad libitum weight levels. The present findings suggest that when animals need to be food deprived, preference should be given to using Long-Evans or Sprague-Dawley rats. If Fischer rats must be used, they should not be deprived below 90-95% of their ad libitum body weight. Strategies for future food deprivation studies are discussed, as well as a comparison of the effects of mild and severe food deprivation. PMID- 11495660 TI - Repeated cocaine treatment activates flank marking in adolescent female hamsters. AB - Cocaine abuse during adolescence represents a significant health risk due to the potential for both acute and long-term negative physical and psychological sequelae, including increased aggressive behavior. This study examined the effect of adolescent cocaine treatment on flank marking (i.e., a stereotypic motor behavior that is part of the response pattern of offensive aggression) in female and male Syrian hamsters (Mesocricetus auratus). Adolescent cocaine treatment activated flank marking in female hamsters when animals were measured upon return to their home cage immediately following drug treatment. Sex differences were observed in cocaine-induced flank marking, as males failed to flank mark when returned to the home cage. In females, the behavioral response was most marked on Day 11 of cocaine treatment in all doses tested. Yet, animals treated with low dose cocaine (0.5 mg/kg/day) showed the most significant increase in flank marking on and from Day 11 forward as compared to medium- and high-dose cocaine treated animals and controls. In addition, the response of cocaine-treated animals was vigorous and nearly immediate, as >75% of the flank marks scored were performed within the first 2 min of the behavioral test in >85% of animals examined. Measures of locomotion showed that cocaine had stimulatory effects on motor activity in adolescent female hamsters at all doses tested. Cocaine-treated animals did not differ in body weight gain from controls, suggesting no dramatic physiological effects of adolescent cocaine exposure on body growth at the doses tested. PMID- 11495661 TI - Development of brief methods to classify individuals by PROP taster status. AB - The purpose of this study was to develop brief methods for classifying individuals by genetic taste sensitivity to 6-n-propylthiouracil (PROP). Two methods are described, which are modifications of a commonly used, suprathreshold procedure. Eighty-nine adult subjects rated the perceived intensity of solutions of 0.032, 0.32 and 3.2 mmol/l PROP and 0.01, 0.1, 1.0 mol/l sodium chloride (NaCl) (three-solution test), as well as solutions of 0.32 mmol/l PROP and 0.1 mol/l NaCl (one-solution test) using the Labeled Magnitude Scale (LMS). Subjects were classified as PROP nontasters (n=22), medium tasters (n=51) or supertasters (n=16) by the three-solution test. Taster status was independently determined by the one-solution test using numerical cutoff scores, which were determined by calculating the +/-95% confidence interval around the group means for PROP taste intensity. Supertasters gave PROP a rating of > or =51 ("very strong" on the LMS) and nontasters gave PROP a rating of < or =15.5 (approximately "moderate" on the LMS). Medium tasters fell between these two limits. Ninety-one percent of nontasters, 82% of medium tasters and 89% of supertasters were classified in a similar way by the two methods. Agreement between methods was high [coefficient of association (P)=0.74; P < or =.001]. These data suggest that three- and one solution methods can reliably classify subjects by taste sensitivity to PROP and could provide valuable tools in population-based studies. PMID- 11495662 TI - Dihydrotestosterone activates sexual behavior in adult male hamsters but not in juveniles. AB - The effect of an androgenic metabolite of testosterone, dihydrotestosterone (DHT), on reproductive behavior and brain androgen receptor (AR) immunoreactivity was compared in juvenile and adult male Syrian hamsters. Prepubertal and adult animals were castrated and treated with 0, 500, or 1000 microg of DHT daily for 1 week and then tested for their ability to engage in mating behavior. The 1000 microg dose of DHT activated intromissions in adult but not prepubertal males. Brains were collected immediately after the behavioral test to investigate whether the lack of a behavioral response to DHT prior to puberty is associated with fewer AR-immunoreactive (AR-ir) cells in the forebrain nuclei that mediate male sexual behavior. In four of the five nuclei within the behavioral circuit that were examined, the number of AR-containing cells was similar in prepubertal and adult males treated with 1000 microg of DHT. Only in the anterior medial amygdala (MeA) was there a greater number of AR-ir cells in adults. These data indicate that (1) DHT does not activate components of male reproductive behavior prior to puberty and (2) the lack of behavioral responsiveness to DHT in prepubertal males is most likely not related to an overall reduction in ARs within the forebrain circuit that mediates mating behavior. PMID- 11495663 TI - Cognitive demand and blood glucose. AB - Previous research has identified that glucose administration can enhance cognitive performance, especially during more intense cognitive processing. There appears to be a reciprocal relationship between falling glucose levels and cognitive performance, particularly under conditions of cognitive demand. The present placebo-controlled, double-blind, balanced, crossover study examined the possibility that a high cognitive load may produce changes in blood glucose levels. A secondary aim was to examine the effects of glucose on tasks of varying cognitive demand load. The effects of a glucose drink on participants' performance of a serial subtraction task (computerised Serial Sevens), a somatically matched control task (key-pressing), a short interval Word Memory task and a Word Retrieval (Verbal Fluency) task were assessed. The change in blood glucose during the demanding computerised Serial Sevens was compared to the change occurring during the key-pressing control. Glucose consumption significantly improved performance on Serial Sevens, with a trend for improved performance on Word Retrieval and no effect on the Word Memory task. Compared with the control task, Serial Sevens resulted in a significant reduction in blood glucose in both drink conditions. This accelerated decay was significantly greater following glucose than placebo. It is suggested that the amount of cognitive load associated with task performance is an index of its sensitivity to enhancement by glucose. Furthermore, a period of intense cognitive processing leads to a measurable decrease in levels of peripherally measured blood glucose, which may be linked to increased neural energy expenditure. However, the relative contribution of central and peripheral (e.g. cardiac) activity to this effect has yet to be determined. PMID- 11495664 TI - Conditioned food aversion in Suncus murinus (house musk shrew) - a new model for the study of nausea in a species with an emetic reflex. AB - The lack of a small animal model with an emetic reflex in which the relationship between conditioned food aversion and emesis could be investigated prompted a study of the insectivore, Suncus murinus (the house musk shrew). A novel food (either tuna or chicken cat food) was paired (C+) with a single exposure to either nicotine (4 mg/kg sc), motion (1 Hz, 4 cm, 10 min) or lithium chloride (100 mg/kg ip) or was paired (C-) with either saline or sham exposure to motion. Nicotine and motion both induced emesis (retching/vomiting) but lithium chloride did not. All three treatments produced a conditioned food aversion after a single pairing with consumption of C+ food. When given a choice between the two foods, S. murinus given lithium chloride, motion exposure and nicotine consumed, respectively, only 25%, 23% and 1% of their total intake from the C+ food. This study shows that a conditioned food aversion can be readily induced in S. murinus and that the induction of emesis can be uncoupled from food aversion. S. murinus provides a promising new model in which the relationship between emesis, nausea and conditioned food aversion can be investigated. PMID- 11495665 TI - Overexpression of agouti protein and stress responsiveness in mice. AB - Ectopic overexpression of agouti protein, an endogenous antagonist of melanocortin receptors' linked to the beta-actin promoter (BAPa) in mice, produces a phenotype of yellow coat color, Type II diabetes, obesity and increased somatic growth. Spontaneous overexpression of agouti increases stress induced weight loss. In these experiments, other aspects of stress responsiveness were tested in 12-week-old male wild-type mice and BAPa mice. Two hours of restraint on three consecutive days produced greater increases in corticosterone and post-stress weight loss in BAPa than wild-type mice. In Experiment 2, anxiety type behavior was measured immediately after 12 min of restraint. This mild stress did not produce many changes indicative of anxiety, but BAPa mice spent more time in the dark side of a light-dark box and less time in the open arms of an elevated plus maze than restrained wild-type mice. In a defensive withdrawal test, grooming was increased by restraint in all mice, but the duration of each event was substantially shorter in BAPa mice, possibly due to direct antagonism of the MC4-R by agouti protein. Thus, BAPa mice showed exaggerated endocrine and energetic responses to restraint stress with small differences in anxiety-type behavior compared with wild-type mice. These results are consistent with observations in other transgenic mice in which the melanocortin system is disrupted, but contrast with reports that acute blockade of central melanocortin receptors inhibits stress-induced hypophagia. Thus, the increased stress responsiveness in BAPa mice may be a developmental compensation for chronic inhibition of melanocortin receptors. PMID- 11495666 TI - Cross-modal transfer effects on visual discrimination depends on lesion location in the rat visual system. AB - The effects of postoperative visual and auditory training on a brightness discrimination task were examined after lesions of various structures in the visual system. In Experiment 1, rats were trained to avoid shock with visual intensity cues. Twenty-four hours later, each rat received bilateral lesions in one of the following areas of the visual system: (1) sham, (2) visual cortex (VC), (3) pretectal (PT) area, (4) combined PT/VC, (5) superior colliculus (SC), or (6) combined SC/VC. Six days later, each rat received either training with visual or auditory intensity cues, or no training. The next day all rats were retrained on the preoperative visual avoidance task. All lesions except those in the SC condition produced relearning deficits. Auditory training reduced these deficits significantly more than visual training, except in rats with combined SC/VC lesions. In Experiment 2, sham and combined PT/VC lesion rats were given either direct or reversal intensity training using visual or auditory cues before relearning the visual discrimination. Rats given auditory direct training relearned the task faster than rats given reversal training or visual direct training. Postinjury training with an intact sensory system can enhance functional recovery more effectively than training with the damaged system. The differential effects of direct and reversal training suggest that cross-modal training involves both specific and nonspecific transfer that may be mediated through the VC or the SC. PMID- 11495667 TI - Polygraphic investigation of 24-h waking distribution in infants. AB - An evening forbidden zone for sleep has been shown for adults. This research was aimed to ascertain the prevalence of waking in the evening in early development. Twelve infants, aged between 2 weeks and 11 months 3 weeks, were each recorded once over a 24-h period. Recordings included EEG, EOG, EMG, respiration, ECG, and behavioural observation. Four states were defined by combining behavioural and electrophysiological data. Wakefulness was defined by the presence of eyes open, eye movements and additionally body movements, and irregular respiration. In infants younger than 12 months and a half, a greater amount of wakefulness was observed in the time interval between 17 and 20. Older infants show uniform high amount of wakefulness during daytime. Our data corroborate the hypothesis that evening hours are those most frequently characterised by the behavioural waking state, suggesting that a forbidden zone for sleep exists in development and that its time placement corresponds to the one observed in the adult. PMID- 11495668 TI - Demand-feeding rhythm in rainbow trout and European catfish. Synchronisation by photoperiod and food availability. AB - The effect of light-dark (LD) cycle and food availability was tested on the demand-feeding rhythm of single and groups of rainbow trout and European catfish. Under LD and free food access, most trout and catfish displayed, respectively, a diurnal and a nocturnal pattern of demand-feeding activity, whereas a few fish or groups of fish switched from diurnalism to nocturnalism or vice versa. In both species held under constant lighting conditions and a restricted feeding (RF) cycle (RF 20:4), the demand-feeding rhythm rapidly synchronised to food availability. The demand-feeding rhythm was under endogenous control and, in rainbow trout, periodogram analysis suggested the existence of two oscillators, one synchronised by photoperiod (LEO) and the other by food (FEO). When submitted to both LD and RF cycles, LD was, at least in the rainbow trout, the dominant zeitgeber synchronising the demand-feeding rhythm. In catfish, food availability rapidly synchronised demand-feeding rhythm. Finally, in both species, the synchronisation of single fish to LD or feed availability appeared slower than that of groups of fish, supporting the idea that social organisation affects the circadian activity in fish. PMID- 11495669 TI - Sex differences in olfactory self-recognition. AB - This study investigated sex differences in the ability to recognize one's own body odor accompanied by an attempt to account for variance in this ability by comparing ratings of self-body odor and other odors on a visual analog scale (VAS). Whereas over half (59.4%) of the females were able to identify their own odor, only one out of 18 (5.6%) males were able to recognize their own odor. Females rated their own secretions as significantly lower on a pleasant-positive factor than males rated their own odors (axillary secretions), but there was no difference in ratings between those who could and those who could not identify their own odor. The dimensions tapped by the VAS used in this study do not seem to account for the ability to identify one's own body odors. PMID- 11495670 TI - Brain catalase activity is highly correlated with ethanol-induced locomotor activity in mice. AB - It has been demonstrated that acute administration of lead to mice enhances brain catalase activity and ethanol-induced locomotion. These effects of lead seem to be related, since they show similar time courses and occur at similar doses. In the present study, in an attempt to further evaluate the relation between brain catalase activity and lead-induced changes in ethanol-stimulated locomotion, the interaction between lead acetate and 3-amino-1H,2,4-triazole (AT), a well-known catalase inhibitor, was assessed. In this study, lead acetate or saline was acutely injected intraperitoneally to Swiss mice at doses of 50 or 100 mg/kg 7 days before testing. On the test day, animals received an intraperitoneal injection of AT (0, 10, or 500 mg/kg). Five hours following AT treatment, ethanol (0.0 or 2.5 g/kg, ip) was injected and the animals were placed in open-field chambers, in which locomotion was measured for 10 min. Neither lead exposure nor AT administration, either alone or in combination, had any effect on spontaneous locomotor activity. AT treatment reduced ethanol-induced locomotion as well as brain catalase activity. On the other hand, ambulation and brain catalase activity were significantly increased by both doses of lead. Furthermore, AT significantly reduced the potentiation produced by lead acetate on brain catalase and on ethanol-induced locomotor activity in a dose-dependent manner. A significant correlation was found between locomotion and catalase activity across all test conditions. The results show that brain catalase activity is involved in the effects of lead acetate on ethanol-induced locomotion in mice. Thus, this study confirms the notion that brain catalase provides the molecular basis for understanding some of the mechanisms of the action of ethanol in the central nervous system. PMID- 11495671 TI - Environmental enrichment: effects on spatial memory and hippocampal CREB immunoreactivity. AB - Environmental enrichment has been shown to improve performance in tests of spatial memory, induce neurogenesis in the hippocampus, enhance survival of newly formed granule cells, and inhibit spontaneous apoptosis. Although neuroplasticity of the mammalian brain declines with age, recent evidence suggests that the adult brain exhibits significant plasticity in response to environmental stimulation. The present study was designed to evaluate the effect of environmental enrichment on spatial memory and on immunoreactivity to cAMP response element binding protein (CREB) from the hippocampus. C57/BL/6 mice were trained in a Morris water maze after exposure to an enriched environment, either from 35 to 94 days or from 100 to 159 days of age. Hippocampal tissue from representative animals was later analyzed by Western blot for CREB immunoreactivity. Results indicate that environmental enrichment (particularly during the earlier period) improved performance on the Morris water maze and tended to increase immunoreactivity to CREB in the hippocampus. Social interaction by itself did not result in significant differences in navigational performance. Results with regard to social interaction and CREB immunoreactivity were mixed. Results are discussed in terms of evaluating the construct of enrichment, the correlation of CREB transcription and behavior change, and the importance of the developmental period for enrichment. PMID- 11495672 TI - The effect of Ginkgo biloba on memory in healthy male volunteers. AB - The purpose of this study was to investigate possible effects of Ginkgo biloba, a widely used herbal extract, on memory. This study incorporated a double-blind, placebo-controlled design, which used 30 healthy male subjects in each of two groups. The treatment group received two 60-mg tablets of BioGinkgo (27/7) [corrected] daily for 5 days, while the placebo group received a placebo. On the fifth day, after a 2-h waiting period, all subjects were given the Sternberg Memory Scanning Test [Q. J. Exp. Psychol. 27 (1975) 1.], a reaction time control test, the vocabulary and digit span subtests of the WAIS-R [Wechsler D. Manual for the Wechsler adult intelligence scale - revised. New York: Psychological Corporation, 1981.], a reading span test [J. Verbal Learn. Verbal Behav. 19 (1980) 450.] and a prose recall test [Discourse Proc. 13 (1990) 387.]. Blood pressure, heart rate and side effects were also monitored throughout the study. Nonsignificant results were found on all interactions involving treatment group on all tests except the Sternberg Memory Scanning Test. The extract appeared to be safe but largely ineffective in enhancing memory. PMID- 11495673 TI - Abstracts of the International Society of Neuroimmunology 6th International Congress. Edinburgh, Scotland, 3-7 September 2001. PMID- 11495674 TI - Tachykinins as modulators of the micturition reflex in the central and peripheral nervous system. AB - In the normal urinary bladder, tachykinins (TKs) are expressed in a population of bladder nociceptors that is sensitive to the excitatory and desensitizing effects of capsaicin (i.e., capsaicin-sensitive primary afferent neurons (CSPANs)). Several endobiotics or xenobiotics excite CSPANs and release TKs and other mediators at both the peripheral and spinal cord level. The peripheral release of TKs determines a set of responses (known as neurogenic inflammation) that includes vasodilatation, plasma protein extravasation, smooth muscle contraction and stimulation of afferent nerves. Following chronic inflammation, both immune cells and capsaicin-resistant sensory neurons can de novo express TKs: whether these pools of TKs are releasable and contribute to inflammatory processes is presently unsettled. At the spinal cord level, the release of TKs contributes in determining an altered pattern of vesicourethral reflexes in response to nociceptive stimulation of the bladder by conveying: (a) the afferent transmission to supraspinal sites, and (b) descending or sensory inputs to the sacral parasympathetic nucleus (SPN). Recent evidence also attribute a synergetic role of TKs in the supraspinal modulation of the sensory arm of the micturition reflex. The overall available information suggests that TK receptor antagonists may affect bladder motility/reflexes which occur during different pathological states, while having little influence on the normal motor bladder function. PMID- 11495675 TI - Parathyroid hormone-related protein overexpression in the human colon cancer cell line HT-29 enhances adhesion of the cells to collagen type I. AB - The present experiments examined the potential ability of parathyroid hormone related protein (PTHrP) to influence growth of the human colon cancer cell HT-29 and the ability of the cell to adhere to several extracellular matrix (ECM) proteins found in normal tissues. Addition of PTHrP analogs, PTHrP (1-34), PTHrP (67-86), or PTHrP (107-139), to HT-29 cells in culture did not influence cell growth or the adhesion of the cells to wells coated with fibronectin, laminin, or collagen type I. Likewise, in HT-29 cells induced to overexpress PTHrP by stable transfection with PTHrP cDNA, compared to vector-transfected control HT-29 cells, no effect on cell growth occurred. However, in the transfected cells, the increased production of PTHrP significantly enhanced cell adhesion to type I collagen but not to fibronectin or laminin. The results raise the possibility that PTHrP might play a role in colon tumor invasion and metastasis by influencing cell adhesion to specific extracellular matrix proteins. PMID- 11495676 TI - Identification of CCK-B/gastrin receptor splice variants in human peripheral blood mononuclear cells. AB - There is increasing evidence for a direct interaction of the enteric nervous and immune system. Receptors for neuropeptides such as VIP, somatostatin, and substance P have been characterised in human immuno-haematopoietic cells but little is known about the functional significance and expression of receptors for cholecystokinin (CCK) on cells of the immune system. There are only few studies that describe the expression of CCK receptors on human leukaemia-derived cell lines but the receptor structure and function in normal leukocytes have not been clearly established. We therefore sought to determine CCK receptor expression, structure, and function in nontransformed human peripheral blood mononuclear cells.Full-length cDNA clones encoding the human CCK-A and CCK-B/gastrin receptor are expressed in peripheral blood mononuclear cells from healthy volunteers without haematopoietic malignancy. In addition to wild-type CCK-B/gastrin receptor cDNAs, we isolated a splice variant with an in frame insertion of 69 amino acids within its putative third intracellular receptor loop. Dideoxy sequence analysis revealed that the cDNA of this splice variant comprises exons 1 4 but retains intron 4 (207 bp) in the absence of mutations within the splice donor sites. Transient expression of this splice variant in COS-7 cells reveals wild-type affinity for CCK-8, Gastrin-17, and antagonist L-365,260. Affinity for glycine-extended gastrin-17 was not increased when compared to the wild-type CCK B/gastrin receptor. In vitro, gastrin decreased 3H-thymidine labelling in phytohaemagglutinin-pretreated mononuclear cells at a half-maximally effective concentration of 1.5 nM. We also isolated a cDNA encoding another splice variant of the CCK-B/gastrin receptor with a 158 bp deletion of the entire exon 4 sequence. We conclude that wild-type transcripts of both CCK receptor subtypes and splice variants of the CCK-B/gastrin receptor are expressed in nontransformed human mononuclear cells and that gastrin exhibits antiproliferative effects. PMID- 11495677 TI - Secretion of the trefoil factor TFF3 from the isolated vascularly perfused rat colon. AB - The trefoil factor TFF3 is a peptide predominantly produced by mucus-secreting cells in the small and large intestines. It has been implicated in intestinal protection and repair. The mechanisms that govern TFF3 secretion are poorly understood. The aim of this study was, therefore, to evaluate the influence of neurotransmitters, hormonal peptides and mediators of inflammation on the release of TFF3. For this purpose, an isolated vascularly perfused rat colon preparation was used. After a bolus administration of 1 ml isotonic saline into the lumen, TFF3 secretion was induced by a 30-min intra-arterial infusion of the compounds to be tested. TFF3 was evaluated in the luminal effluent using a newly developed radioimmunoassay. TFF3 was barely detected in crude luminal samples. In contrast, dithiothreitol (DTT) treatment of the effluent revealed TFF3 immunoreactivity, which amounted to about 0.3 pmol min(-1) cm(-1) in the basal state. Gel chromatography of DTT-treated luminal samples revealed a single peak that co eluted with the monomeric form of TFF3. TFF3 was not detected in the portal effluent. Bethanechol (10(-6)-10(-4) M), vasoactive intestinal peptide (VIP, 10( 8)-10(-7) M) or bombesin (10(-8)-10(-7) M) induced a dose-dependent release of TFF3. In contrast, substance P evoked a modest release of TFF3, whereas calcitonin gene-related peptide (CGRP), somatostatin, neurotensin or peptide YY (PYY) did not modify TFF3 secretion. The degranulator compound bromolasalocid, 16,16-dimethyl PGE2 (dmPGE2) or interleukin-1-beta (IL-1-beta) also evoked a marked release of TFF3. In conclusion, TFF3 in the colonic effluent is present in a complex. This association presumably involves a disulfide bond. Additionally, the present results suggest a role for enteric nervous system and resident immune cells in mediation of colonic TFF3 secretion. PMID- 11495678 TI - Regulation of apoptosis by somatostatin and substance P in peritoneal macrophages. AB - Recent studies have shown that somatostatin (SOM) inhibits interleukin 6 (IL-6) and interferon gamma (IFNgamma) production by lymphocytes and peritoneal macrophages, whereas substance P (SP) enhances these cytokines production. To define the mechanism of the cytokine production enhancements and inhibitions by SOM and SP, we examined the expression of apoptosis modulator, p53, Bcl-2, Bax, inducible nitric oxide synthase (iNOS), Fas, caspase-8 and nitric oxide (NO) in thioglycolate-elicited peritoneal macrophages. SOM caused up-regulation of p53, Bcl-2, Fas and caspase-8 activities, and down-regulation of iNOS expression and NO production. On the other hand, SP slightly induces p53 and highly induces Bcl 2, iNOS expression and NO production. These data suggest that apoptosis by SOM may occur by a Bax- and NO-independent p53 accumulation, and through Fas and caspase-8 activation pathways, and that the inducible expression of Bcl-2 and NO production by SP may contribute to prevent the signals of apoptosis by Bax, and via Fas and caspase-8 activation. PMID- 11495679 TI - Location of renin-angiotensin system components in the hypoglossal nucleus of the rat. AB - The renin-angiotensin system (RAS) in the hypoglossal nuclei of the rat was studied by immunohistochemistry. Antibodies to angiotensin AT(1) receptor (AT1), angiotensinogen (ANG), renin (REN), angiotensin converting enzyme (ACE) and angiotensin II (AII) were used. All the components of the RAS with the exception of renin were detected. Light and electron microscopy revealed the following results: ANG was predominantly found in astrocytes, with small amounts in neuronal dendrites; ACE was found in the cytoplasm of neurons, dendrites and astrocyte processes; AT1 was found in the cytoplasm of neurons and dendrites, but not on the membrane; and AII was found mainly in astrocytes with some located in the dendrites and cytoplasm. Right hypoglossal nerve lesion caused an increase in expression of AT1 in neurons as early as 2 days post-lesion. An increase in expression of ANG in astrocytes was also seen, but at a much later time of 3 weeks post-lesion. For AII, staining occurred in both the neurons and astrocytes in the undamaged hypoglossal nucleus. Nerve lesion caused a disappearance of neuronal stains and an increase in astrocyte stains. There were no changes in ACE staining after nerve lesion. We speculate that ANG and AII are made within the astrocytes, whereas ACE could either be uptaken from blood or de novo synthesized. AT1 may potentially be internal soluble receptors. As to the function of AII in the hypoglossal nucleus, the data do not support AII as a neurotransmitter in the hypoglossal nucleus. It may function as a neuromodulator and also be involved in basic cellular activities, e.g. regulation of transcription factors. PMID- 11495680 TI - Interactions between orexin A, NPY and galanin in the control of food intake of the goldfish, Carassius auratus. AB - The neuropeptides orexin A (OXA), neuropeptide Y (NPY) and galanin (GAL) have been shown to play a role in the regulation of food intake in mammals. They also significantly stimulate feeding in goldfish. In order to assess the interactions between these peptides in the control of feeding in goldfish, we investigated the effects of central injection of specific receptor antagonists for NPY (BIBP 3226) and GAL (M40) on OXA-induced feeding and the effects of desensitization of orexin receptors on NPY- and GAL-induced feeding. We investigated the effects of BIBP 3226 on GAL-induced feeding and the effects of M40 on NPY-induced feeding. We also examined the effects of coinjection of each pair of neuropeptides on feeding behavior. Injections of 10 ng/g OXA, 5 ng/g NPY and 10 ng/g GAL each induced an increase in feeding. Fish treated with 5 ng/g BIBP or 20 ng/g M40 had food consumption similar to saline controls. BIBP at 5 ng/g significantly reduced NPY- and OXA-induced feeding. Injections of 20 ng/g M40 significantly decreased GAL induced feeding, but had no effect on OXA-induced feeding. Blocking of orexin receptors by treatment with high doses of OXA (100 ng/g) resulted in a decrease in both NPY- and GAL-induced feeding. Coinjection with 0.5 ng/g OXA and either 0.5 ng/g NPY or 0.5 ng/g GAL resulted in a food intake higher than that observed in saline control fish and in fish treated with NPY or GAL alone at 0.5 ng/g. NPY mRNA expression was increased in the telencephalon and in the hypothalamus compared to saline-treated fish, following injection of OXA. These results indicate that both NPY and GAL are at least, in part, dependent on coaction with OXA for the stimulation of food intake and feeding behavior in goldfish. In addition, the effects of OXA are mediated, in part, by the NPY pathway. This suggests a functional interdependence between these three peptidergic systems in the control of energy balance in goldfish. PMID- 11495681 TI - Neuropeptide Y effects on murine natural killer activity: changes with ageing and cAMP involvement. AB - Changes in the bidirectional interaction between the nervous and the immune systems have been proposed as a cause of ageing. Neuropeptides, such as neuropeptide Y (NPY), could show different effects on immune function with age. In the present work, we have studied the in vitro action of a wide range of NPY concentrations, i.e. from 10(-13) to 10(-7) M, on natural killer (NK) activity, a function which decreases with age. Spleen, axillary nodes, thymus and peritoneum leukocytes from mice of different ages: young (12+/-2 weeks), adult (24+/-2 weeks), mature (50+/-2 weeks) and old (72+/-2 weeks) were used. Stimulation by NPY of NK activity was observed in adult and mature animals in axillary nodes and thymus, and an inhibition in the spleen from young mice. The specificity of the NPY effect on cytotoxic activity was confirmed using a C-terminal fragment of NPY. Furthermore, cAMP levels in leukocytes were found to be decreased by NPY in adult mice, suggesting an involvement of this messenger system in the NK modulation by this neuropeptide. PMID- 11495682 TI - Blockade effects of (Nphe1)Nociceptin(1-13)-NH(2) on anti-nociception induced by intrathecal administration of nociceptin in rats. AB - The present study investigated the roles of the opioid-receptor-like (ORL1) receptor and its endogenous ligand nociceptin on nociception in the spinal cord of rats. Intrathecal administration of 10 nmol of nociceptin produced significant increases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation. There were no significant changes of average maximum angles in inclined plane tests after intrathecal injection of 10 nmol of nociceptin in rats. The intrathecal nociceptin-induced increases in HWL were antagonized by intrathecal administration of (Nphe1)Nociceptin(1-13)-NH(2), a selective antagonist of ORL1 receptor, in a dose-dependent manner. The results demonstrated that ORL1 receptor is involved in the nociceptin-induced anti-nociceptive effect in the spinal cord of rats. PMID- 11495683 TI - Progesterone and norethisterone have different effects on tachykinin-like immunoreactivity in rat cortex and striatum. AB - OBJECTIVE: The purpose of this study was to investigate the effects of progesterone and the most commonly prescribed synthetic progestogen, norethisterone, on regional immune-like reactivity of neuropeptide Y (NPY), substance P (SP), neurokinin A (NKA) and neurotensin (NT) in brains of female ovariectomized estradiol-substituted rats. RESULTS: Norethisterone+estradiol treated rats had 44% lower SP levels compared with estradiol-only-treated in frontal cortex and 20% lower NKA levels in comparison with progesterone+estradiol treated in frontal cortex. Progesterone+estradiol-treated rats had 66% lower SP levels in striatum in comparison with both estradiol-only-treated and norethisterone+estradiol-treated. No significant results were found for NPY and NT. CONCLUSION: Progesterone and the synthetic progestogen, norethisterone, have different effects on SP- and NKA-like immunoreactivity in rat cortex and striatum. The effects of NET on SP- and NKA-like immunoreactivity in frontal cortex may contribute to the mood effects ascribed to this progestogen in clinical usage. PMID- 11495684 TI - Expression of receptors regulating gastric acidity in the developing sheep stomach. AB - Acid secretion first appears in the stomach during the later stages of fetal development. Gastric acid secretion is regulated by the stimulatory effects of gastrin, histamine, acetylcholine and the inhibitory actions of somatostatin on their respective receptors. A semi-quantitative reverse transcriptase-polymerase chain reaction method for the determination of changes in mRNA expression for these receptors was developed and correlated with known changes in gastric acidity. Glyceraldehyde-3-phosphate dehydrogenase (GAP-DH) was used as a reference and an internal standard. The antrum and fundus from four age groups were assayed: 80 days of gestation, 110 days of gestation, term (145 days) and adult animals. The CCK B/gastrin and the histamine (H(2)) receptor mRNA were significantly lower in samples from the fundus of fetuses, from 80 and 110 days of gestation when compared with the adult fundus. Histamine receptor mRNA in the antrum was also significantly lower in the 80 and 110 days of gestation samples relative to the term fetal antrum. Somatostatin II receptor mRNA levels in the antrum decreased with increasing age with no change in the fundus. These findings suggest that changes in receptor gene expression, may be responsible for the diminished gastric acidity and responsiveness observed in the fetal stomach. PMID- 11495685 TI - Modulatory effect of two novel CGRP receptor antagonists on nasal vasodilatatory responses to exogenous CGRP, capsaicin, bradykinin and histamine in anaesthetised pigs. AB - Calcitonin gene-related peptide (CGRP) is a 37-amino acid peptide and potent vasodilatator agent located in sensory C fibres. Several functional studies suggest that CGRP could be involved in the vasodilatation of different vascular beds during neurogenic inflammation. We have studied, in pentobarbital anaesthetised pigs, the antagonistic effect of local intra-arterial (i.a.) pretreatment with the analogues CGRP 8-37, [D31, P34, F35]CGRP 27-37 and [N31, P34, F35]CGRP 27-37 on the vasodilatation of the nasal vascular bed induced by exogenous CGRP, capsaicin, bradykinin (BK) and histamine. The attenuating effect of CGRP 8-37 analogue on exogenous CGRP-induced vasodilatation, previously described in other in vivo animal models, was confirmed in the pig nasal mucosa. It also interfered with BK-and, to a lesser extent, with capsaicin-and histamine induced decrease in vascular resistance. CGRP 27-37 analogues reduced the duration of CGRP-, capsaicin- and BK-induced vasodilatation by more than 50%. Peak values of vasodilatation were attenuated by more than 25% overall. Attenuation of histamine-induced decrease in vascular resistance was less pronounced. It is concluded that CGRP 27-37 analogues antagonise the action of exogenous CGRP, capsaicin, BK and histamine by attenuating their vasodilatation effect, both in intensity and duration. These results strongly suggest that BK- and histamine-induced vasodilatation is partly mediated by CGRP. CGRP 8-37 and 27 37 appear to be potential contributors to the study of CGRP and its physiological role in neurogenic inflammation. In addition, they may have putative therapeutic applications in the treatment of rhinitic patients suffering from chronic nasal obstruction. PMID- 11495686 TI - PACAP 1-38 as neurotransmitter in the porcine antrum. AB - The concentration of PACAP 1-38 in porcine antrum amounted to 15.4+/-7.9 and 20.3+/-8 pmol/g tissue in the mucosal and muscular layers. PACAP immunoreactive (IR) fibres innervated the muscular (co-localised with VIP) and submucosal/mucosal layers (some co-storing VIP and CGRP) including myenteric and submucosal plexus and blood vessels. Only myenteric nerve cell bodies contained PACAP-IR (co-storing VIP). In isolated perfused antrum, vagus nerve stimulation (8 Hz) and capsaicin (10(-5) M) increased PACAP 1-38 release. PACAP 1-38 (10(-9) M) increased substance P (SP), gastrin releasing peptide (GRP) and VIP release. PACAP 1-38 (10(-8) M) inhibited gastrin secretion and stimulated somatostatin secretion and motility dose-dependently. PACAP-induced motility was strongly inhibited by the antagonist PACAP 6-38 but also by atropine and substance P antagonists (CP99994/SR48968) but PACAP 6-38 had no effect on vagus-induced secretion or motility. CONCLUSION: PACAP 1-38 may be involved in antral motility and secretion by interacting with cholinergic, SP-ergic, GRP-ergic and/or VIP ergic neurones, and may also be involved in afferent reflex pathways. PMID- 11495687 TI - [D-LEU-4]-OB3, a synthetic leptin agonist, improves hyperglycemic control in C57BL/6J ob/ob mice. AB - We have recently shown that the activity of a synthetic peptide corresponding to amino acid residues 116-130 of secreted mouse leptin is contained in a restricted sequence at the amino terminus of the peptide, between residues 116-122 (Ser-Cys Ser-Leu-Pro-Gln-Thr, OB3). Substitution of the Leu residue at position 4 of OB3 with its D-isomer ([D-Leu-4]-OB3) enhanced the ability of OB3 (1 mg/day, ip, 7 days) to reduce body weight gain, food and water intake, and serum glucose in female C57BL/6J ob/ob mice. In the present study, we have utilized a pair-feeding approach to demonstrate that the antihyperglycemic action of [D-Leu-4]-OB3 is not solely due to its effects on caloric intake. One group of female C57BL/6J ob/ob mice (n=6) was fed ad libitum, and two additional groups (n=6 per group) were allowed 3.0 g food/mouse daily, an amount previously determined to satisfy [D-Leu 4]-OB3-treated mice. At the end of the 7-day test period, vehicle-injected mice fed ad libitum were approximately 10% heavier than their initial body weights, while pair-fed mice injected with vehicle and [D-Leu-4]-OB3-treated mice lost 5% of their initial body weights. After 1 day of treatment, blood glucose was reduced by 20% in pair-fed vehicle-injected mice, and by 40% in mice given [D-Leu 4]-OB3. Food restriction reduced blood glucose throughout the 7-day study, but not to levels seen in wild-type nonobese C57BL/6J mice of the same sex and age. After 2 days of treatment with [D-Leu-4]-OB3, however, blood glucose was reduced to levels comparable to those seen in wild-type nonobese mice. [D-Leu-4]-OB3 also lowered serum insulin levels by 53% when compared to mice fed ad libitum. Neither pair-feeding nor [D-Leu-4]-OB3 treatment had any apparent effect on thermogenesis. These results suggest that [D-Leu-4]-OB3 exerts its effects on serum glucose not only by suppressing caloric intake, but also through a separate effect on glucose metabolism which may involve increased tissue sensitivity to insulin. PMID- 11495688 TI - Structure-activity and possible mode of action of S-Iamide neuropeptides on identified central neurons of Helix aspersa. AB - Intracellular recordings were made from identified neurons from the suboesophageal ganglia of Helix aspersa. The inhibitory action of nine S-Iamide peptides was investigated. Structure-activity studies suggest that all act through a common receptor, which normally requires FVRIamide at the C terminal, with a preferred length of seven amino acids. Substitution at the N-terminal with alanine (A), threonine (T), proline (P) or leucine (L) results in little change in potency, suggesting the N-terminal requirements are relatively flexible. Ion substitution experiments suggest that potassium is the main ion involved in the inhibitory response to S-Iamide application. Studies using a range of compounds, which modify second messenger systems, would suggest that S-Iamide peptides may interact with adenylate cyclase. No evidence was found for an interaction with either guanylate cyclase or nitric oxide synthase. PMID- 11495689 TI - Effects of hydrosaline treatments on prolyl endopeptidase activity in rat tissues. AB - Enzymatic cleavage of some peptide hormones, neurotransmitters and neuromodulators could be implicated in the regulation of extra- and intracellular fluid volume and osmolality. Prolyl endopeptidase is known to hydrolyze several peptides, which act on hydromineral balance, such as angiotensins, bradykinin, vasopressin, oxytocin, thyrotropin-releasing hormone, neurotensin and opioids. In this work, we analyzed the effects of certain volume and/or osmotic changes in the activity of the soluble and membrane-bound prolyl endopeptidase in several brain areas, heart, lungs, kidney and adrenal and pituitary glands of the rat. Soluble prolyl endopeptidase activity was higher in the renal cortex of the chronic salt-loaded rats than in the control rats. In the water-deprived and polyethylene glycol-treated rats, heart particulate prolyl endopeptidase was lower than in the control rats. Particulate prolyl endopeptidase was also lower in the adrenal gland of the acute salt-loaded rats and in the brain cortex of the water-loaded rats than in the control rats. Data suggest that tissue-dependent peptide hydrolysis evoked by prolyl endopeptidase activity is involved in the water-electrolyte homeostasis. PMID- 11495690 TI - Immunocytochemical localization of angiotensin II receptor subtypes and angiotensin II with monoclonal antibodies in the rat adrenal gland. AB - Angiotensin II (Ang II), a major regulator of cardiovascular function and body fluid homeostasis, mediates its biological actions via two subtypes of G protein coupled receptors, termed AT(1) and AT(2). The primary goal of this study was to raise monoclonal anti-peptide antibodies specific to angiotensin AT(1)- and AT(2) receptor subtypes and to Ang II itself and using these monoclonal antibodies to determine the intraadrenal localization of AT(1) and AT(2) receptors and Ang II in male adult rats. Immunocytochemistry unambiguously demonstrates a regional colocalization of Ang II and angiotensin II receptors in the adrenal gland. The novel antibodies localized Ang II and the AT(1) receptors to the zona glomerulosa of the cortex and to the medulla whereas AT(2) receptors were limited to the medulla. The specificity of immunostaining was documented by pre-adsorption of the antibody with the immunogenic peptide. Our data underscore that AT(1) appears to mediate most of the physiological actions of Ang II in adrenal. Western blot analysis of rat adrenal protein extracts using AT(1) antibody showed a predominant 73-kDa band and a weaker 97-kDa immunoreactive band corresponding to glycosylated forms of the AT(1) receptor. Immunostaining with anti-AT(2) yielded one major immunoreactive band of 73-kDa size and one additional fainter band of 120 kDa. These antibodies may prove of value in unraveling the subcellular localization and intracellular effector pathways of AT(1) and AT(2). PMID- 11495691 TI - Human alpha-defensin 1 (HNP-1) inhibits adenoviral infection in vitro. AB - Adenoviral gene transfer is a promising tool for direct treatment of cystic fibrosis by local application of the CFTR-gene via the airway. However, various host defense mechanisms reduce the adenoviral infectivity and hereby the success of adenoviral transduction. Twenty-eight of 62 BALs from various patients exerted strong inhibition of adenoviral infection of 293 cells. This soluble activity could be attributed to larger peptides rather than to small molecules. Beside immunoglobulins, certain epithelial cell-derived anti-microbial polypeptides called defensins might be involved. Therefore, we investigated the inhibitory potential of the defensins HNP-1 and HBD-2 on adenoviral infectivity. 293 cells infected with adenovirus-type 5 were treated with both peptides. Compared to control, HNP-1 reduced adenoviral infection by more than 95% if administered at 50 microg/ml, and the IC50-value was 15 microg/ml. In contrast, HBD-2 was much less efficient and did not block adenoviral infection at doses up to 50 microg/ml. Our data demonstrate that the presence of certain polypeptides in the BAL, i.e. the defensin HNP-1, might be the major obstacle for adenoviral gene transfer, particularly in patients with inflammatory diseases. PMID- 11495692 TI - Adrenomedullin and proadrenomedullin N-terminal 20 peptide induce histamine release from rat peritoneal mast cell. AB - Adrenomedullin (ADM)-induced histamine release from rat peritoneal mast cells was investigated. We compared the ability of full-length ADM to induce histamine release to the fragments ADM-(1-25) and ADM-(22-52), as well as proadrenomedullin N-terminal 20 peptide (PAMP). ADM (10(-8) to 10(-5) M) and PAMP (10(-8) to 10(-5) M) dose-dependently increased histamine release from peritoneal mast cell preparations. The effect of ADM-(1-25) was similar to ADM, whereas ADM-(22-52) did not show any effects. These data suggest the relative importance of the ADM C terminal fragment, which contains a six-membered ring structure. Histamine release, induced by ADM, was significantly and dose-dependently inhibited by the addition of ADM-(22-52) (10(-5) M), Ca(2+) (0.5 to 2.0 mM), and benzalkonium chloride (3 to 7 microM), a selective inhibitor of Gi type G proteins. In contrast, PAMP (10(-5) M)-induced histamine release was not inhibited by Ca(2+). These results suggest that ADM induce histamine release via a putative ADM receptor in a manner sensitive to Gi-protein function and extracellular Ca(2+) concentration, and that PAMP might produce its effect by a different mechanism than ADM. PMID- 11495693 TI - The neuropeptide calcitonin gene-related peptide differently modulates proliferation and differentiation of smooth muscle cells in culture depending on the cell type. AB - Calcitonin gene-related peptide (CGRP) is a neuropeptide present around vasculature very early during development, when smooth muscle cells (SMC) are still proliferating and not yet totally differentiated. We investigated the effects of CGRP on proliferation and differentiation of SMC in culture; 10(-7) M CGRP added in the medium of cultured smooth muscle cells every 2 days did not significantly changed cells growth rate in 1% FCS. At the opposite, this treatment modulated proliferation of cells grown in 10% FCS medium. Two distinct populations of SMC with different growth rates were obtained from our primary cultures. SMC which proliferated slowly in the presence of 10% fetal calf serum (FCS) had growth rates positively influenced by CGRP. The quantity of alpha smooth actin expressed by these cells was not influenced by the peptide. On the contrary, SMC which proliferated more rapidly in 10% FCS medium had growth rate inhibited by CGRP. In these cells, CGRP significantly reduced the amount of expressed alpha-smooth actin, an index of SMC differentiation. In both cases, the peptide significantly increased the level of mRNA for all the actin genes. In the light of this dual role of CGRP, it can be presumed that this peptide controls smooth muscle cells proliferation and differentiation in vivo and could thus regulate the homeostasis of the vessel wall. PMID- 11495694 TI - The effect of galanin on wide-dynamic range neuron activity in the spinal dorsal horn of rats. AB - The present study investigated the effect of galanin on wide-dynamic range (WDR) neuron activity in the dorsal horn of the spinal cord of rats. The evoked discharge of WDR neurons was elicited by transdermic electrical stimulation applied on the ipsilateral hindpaw of rats. Galanin was administered directly on the spinal dorsal surface of L3-L5. The evoked discharge frequency of the WDR neurons decreased significantly after the administration of galanin and the effect lasted for more than 30 min. Furthermore, the inhibitory effect of galanin on the evoked discharge frequency of WDR neurons was blocked by following administration of the galanin antagonist galantide, indicating that the inhibitory effect of galanin on the activity of WDR neurons was induced by activating galanin receptors in the dorsal horn of the spinal cord. The results suggest that galanin has an inhibitory role in the transmission of presumed nociceptive information in the dorsal horn of the spinal cord in rats. PMID- 11495695 TI - K(+)-p-nitrophenylphosphatase inhibition by neurotensin involves high affinity neurotensin receptor: influence of potassium concentration and enzyme phosphorylation. AB - Neurotensin (NT), a 13-amino acid peptide, is widely distributed in the brain and peripheral tissues of several mammalian species including man. In adult rat brain NT can bind to two distinct sites, one of high and the other of low affinity, corresponding to NT(1) and NT(2) receptor, respectively; structurally unrelated to these two, a third NT receptor (NT(3)) has been described. We have previously shown that Na(+), K(+)-ATPase is inhibited by NT when using ATP as substrate. In order to determine whether K(+)-stimulated dephosphorylation of this enzyme is involved, we tested NT effect by using p-nitrophenylphosphate, a non-natural substrate. K(+)-p-nitrophenylphosphatase activity was inhibited 42% by NT at 8.6 x 10(-6) M using an incubation medium containing 2 mM KCl but was unaffected in the presence of 5 or 20 mM KCl; however, with such KCl concentrations, NT was enabled to inhibit enzyme activity ( congruent with 35%) provided a suitable ATP:NaCl mixture (0.6:45.0 mM) was added. Mg(2+)-p-nitrophenylphosphatase activity remained unaltered at all conditions tested. Since SR 48692, a selective non-peptide NT(1) antagonist, abolished NT effect, involvement of NT(1) receptor in enzyme inhibition is suggested. PMID- 11495696 TI - Aminopeptidase activities after water deprivation in male and female rats. AB - Aminopeptidases (APs) play a major role in the metabolism of circulating and local peptides, such as angiotensins and vasopressin, substances involved in the control of blood pressure and water balance. In the present work, we studied the influence of dehydration on angiotensinases and vasopressin-degrading activity. Since sex differences may exist in the regulation of water balance by angiotensin II and differential sexual steroid modulation of vasopressin secretion, in response to osmotic stimulation have been reported, gonadotropin releasing hormone (GnRH)-degrading activity was also analysed in serum, neurohypophysis and adrenal glands of male and female rats. Our results did not suggest sex differences in the response to changes in osmolality. GnRH-degrading activity decreased in serum of dehydrated males and females, which suggests a longer action of the peptide under these conditions. In neurohypophysis, there was an increase in the activity of aminopeptidase A (APA), the enzyme responsible for the metabolism of angiotensin II to angiotensin III. This occurs with a decrease in alanyl aminopeptidase activity, which would lead to a prolonged action of angiotensin III by reduction of its metabolism. In adrenals of dehydrated animals, the results would imply a high degree of metabolism of angiotensin III and vasopressin. PMID- 11495698 TI - Yolk-sac hematopoiesis: the first blood cells of mouse and man. AB - OBJECTIVE: To review the process of blood-cell formation in the murine and human yolk sac. DATA SOURCES: Most articles were selected from the PubMed database. DATA SYNTHESIS: The yolk sac is the first site of blood-cell production during murine and human ontogeny. Primitive erythroid cells originate in the yolk sac and complete their maturation, including enucleation, in the bloodstream. Though species differences exist, the pattern of hematopoietic progenitor cell emergence in the yolk sac is similar in mouse and man. In both species, there is a stage of development where both primitive red blood cells and definitive erythroid progenitors are produced in the yolk sac. An "embryonic" hematopoietic stem cell that engrafts in myeloablated newborn but not adult mice can be detected in the murine yolk sac and embryo. Stem-cell activity in the human yolk sac has not been reported. CONCLUSIONS: The yolk sac is the sole site of embryonic erythropoiesis. However, definitive erythroid, myeloid, and multipotential progenitors also originate in the yolk sac. The relationship between these progenitors and the "embryonic" hematopoietic stem cell has not been elucidated. Yolk sac-derived progenitor cells may seed the developing liver via the circulation and serve as the immediate source of the mature blood cells that are required to meet the metabolic needs of the rapidly growing fetus. PMID- 11495699 TI - Prolactin-induced expression of cytokine-inducible SH2 signaling inhibitors in human hematopoietic progenitors. AB - OBJECTIVE: The prolactin (PRL) receptor (PRLR) utilizes the JAK2/STAT-5 pathway and induces expression of cytokine-inducible SH2 (CIS)/JAK2 binding (JAB) signaling inhibitors. We and others recently showed that CIS-3 and JAB abolish PRLR-mediated JAK2 activation and STAT-5 activity, whereas CIS-1, CIS-2, and CIS 4 had a negligible effect. Human CD34(+) hematopoietic progenitors express PRLRs and respond to PRL in vitro by enhanced cytokine-induced colony formation. To assess the signaling mechanism(s) involved in PRL-mediated enhancement of hematopoiesis and to identify further the CIS/JAB targets for PRL-mediated cellular responses, we assayed the effect of PRL, alone or in the presence of interleukin-3 (IL-3), on activation of STAT-5 and expression of CIS/JAB RNA in human cord blood (CB) CD34(+) cells. MATERIALS AND METHODS: Isolated CB CD34(+) cells were incubated in serum-free cultures in the absence or presence of recombinant human (rh)PRL, rhIL-3, or both. Cell lysates were subjected to Western blot analysis with anti-STAT-5 and anti-phospho-STAT-5 antibodies. Isolated RNA was subjected to semiquantitative reverse transcriptase polymerase chain reaction analysis of CIS/JAB expression. RESULTS: STAT-5 tyrosine phosphorylation was similarly induced by PRL and IL-3, with an additive effect detected in the presence of both stimuli. Both PRL and IL-3, alone or combined, failed to induce CIS-3 or JAB RNA expression in CD34(+) cells. Interferon-gamma had no effect on CIS-3/JAB induction in these cells. However, CIS-1 was induced by PRL < IL-3 < PRL+IL-3, whereas CIS-2 expression was induced by PRL = IL-3 < PRL+IL-3. CONCLUSIONS: Our findings show that PRL induces activation of STAT-5 and expression of similar CIS/JAB family members as IL-3 does in human CB CD34(+) cells. Because CIS-1 abolishes STAT-5 activation via the IL-3 but not the PRL receptor, the hematopoietic growth-promoting effects of PRL may involve its capacity to provide sustained STAT-5-mediated stimulatory signals to the cells. PMID- 11495700 TI - Progenipoietin-1: a multifunctional agonist of the granulocyte colony-stimulating factor receptor and fetal liver tyrosine kinase-3 is a potent mobilizer of hematopoietic stem cells. AB - OBJECTIVE: Progenipoietin-1 is an agonist of both the granulocyte colony stimulating factor and fetal liver tyrosine kinase-3 receptors capable of inducing the proliferation of multiple hematopoietic cell lineages. The potential of progenipoietin-1 to mobilize transplantable hematopoietic stem cells into the peripheral blood was evaluated. METHODS: Cohorts of donor mice were treated with either progenipoietin-1, fetal liver tyrosine kinase-3 ligand, granulocyte colony stimulating factor, or a vehicle control. Hematopoietic progenitor/stem-cell activity in donor blood was assayed by radioprotection, multilineage reconstitution, secondary transplantation, and competitive repopulation. RESULTS: Only 1 microL of peripheral blood from progenipoietin-1-treated donors was required to protect 80% of lethally irradiated mice, while in contrast 1 microL of peripheral blood from granulocyte colony-stimulating factor-treated donors failed to protect any recipients. The radioprotected recipients of progenipoietin 1-treated donor cells showed donor-derived (Ly5.2) multilineage hematopoietic reconstitution for up to 6 months. Serial transplantation studies using bone marrow from radioprotected, chimeric recipients demonstrated long-term donor derived hematopoiesis, indicating the successful transplantation of multipotent hematopoietic stem cells. The engraftment potential of progenipoietin-1 donor derived cells was directly compared with donors treated with granulocyte colony stimulating factor or fetal liver tyrosine kinase-3 ligand alone or in combination. Both spleen colony-forming activity and competitive repopulating activity was highest in the blood from progenipoietin-1-treated donors. CONCLUSIONS: These studies demonstrate that progenipoietin-1 is a potent mobilizer of transplantable hematopoietic stem cells and indicate that this dual receptor agonist has greater biologic activity than its constituent molecules. PMID- 11495701 TI - Adenovector-induced expression of human-CD40-ligand (hCD40L) by multiple myeloma cells. A model for immunotherapy. AB - OBJECTIVE: CD40L restores the antigen-presenting cell (APC) function of some B cell tumors and induces professional APC maturation. We therefore evaluated the effects of transgenic CD40L expression on the behavior and immunogenicity of human multiple myeloma (MM) cells. MATERIALS AND METHODS: CD40L expression was induced in a CD40(+) (RPMI 8226) and a CD40(-) (U266B1) human myeloma cell line (HMCL) by adenoviral vector gene transfer. The viability and proliferative activity of control HMCL and HMCL/CD40L were determined by daily trypan blue staining and methyl-3H-thymidine incorporation. Mixed lymphocyte reaction (MLR) with allogeneic mononuclear cells (MNCs) and coculture of allogeneic dendritic cells (DCs) with HMCL expressing transgenic CD40L were used to evaluate the APC function of modified HMCL as well as the role of bystander DCs in inducing an anti-tumor immune response. RESULTS: CD40L expression significantly inhibited the growth of the CD40(+) HMCL and induced apoptosis. These effects were less evident for the CD40(-) HMCL. There was no upregulation of costimulatory molecules on either HMCL following CD40L expression. Both HMCL expressing transgenic CD40L induced maturation of bystander DCs and enhanced their ability to stimulate the proliferation of MNCs. DCs cultured with the poorly immunogenic RPMI 8226 expressing CD40L upregulated T-lymphocyte release of IFN-gamma and other Th1 cytokines (interleukin-2, tumor necrosis factor-alpha). CONCLUSIONS: Our data suggest that transgenic expression of CD40L exerts a dual effect favoring generation of an immune response to human MM. Where the tumor cells are CD40(+), the engagement of CD40 antigen by CD40L on tumor cells induces their apoptosis, allowing uptake of tumor-associated antigen by professional APC. Independently of tumor-cell expression of CD40, transgenic expression of CD40L on tumor cells allows them to stimulate CD40(+) APC, to increase their maturation and their capacity to stimulate cytotoxic T lymphocytes (CTL) that recognize the tumor derived antigens the APC may have engulfed. PMID- 11495702 TI - Additive effect of Apo2L/TRAIL and Adeno-p53 in the induction of apoptosis in myeloma cell lines. AB - OBJECTIVE: We have previously shown that Adenovirus-p53 (Ad-p53) is a potent inducer of apoptosis in myeloma cells expressing nonfunctional p53 and low levels of bcl-2 and that Apo2L/TRAIL is a potent inducer of apoptosis, independent of bcl-2. A study was designed to test the synergy between Ad-p53 and Apo2L/TRAIL in the induction of apoptosis in relation to the expression of DR4/DR5 and DcR1, in cells undergoing Ad-p53-induced apoptosis. METHODS: Replication deficient Ad-p53 and human recombinant Apo2L/TRAIL were used. Myeloma cells with mutated/w.t. p53 and varying expression of bcl-2 were used to test the effect of Ad-p53, Apo2L/TRAIL, or both, on apoptosis, measured by annexin V. RESULTS: Treatment with Ad-p53 resulted in a dose-dependent apoptosis concomitant with a dose dependent increase in the expression of DR4/DR5 and a decrease in the expression of DcR1, in Ad-p53-sensitive cell lines. In these cells, addition of Apo2L/TRAIL to cells treated with Ad-p53 resulted in a dose-dependent increase in apoptosis. Myeloma cells resistant to Ad-p53 had high levels of DR4/DR5 and high levels of DcR1 and treatment with Ad-p53 did not reduce the expression of DcR1. Also, addition of Apo2L/TRAIL to Ad-p53 did not affect the level of apoptosis beyond the level of apoptosis observed with Apo2L/TRAIL alone. CONCLUSIONS: 1) Cotreatment with Ad-p53 and Apo2L/TRAIL resulted in additive apoptosis in myeloma cells expressing nonfunctional p53 and low levels of bcl-2. 2) Resistance to Ad p53 or to the combination of Ad-p53 and Apo2L/TRAIL was not due to the lack of adenovirus receptor (CAR) or low expression of DR4/DR5 but rather due to the relatively high expression of DcR1 receptor. PMID- 11495703 TI - Enforced expression of the GATA-3 transcription factor affects cell fate decisions in hematopoiesis. AB - OBJECTIVE: Three GATA family transcription factors are involved in various aspects of hematopoiesis. Their lineage-restricted expression correlates well with their function in selective lineage commitment and differentiation. We focused on the role of GATA-3 to determine whether an intrinsic variation among different GATA proteins, in addition to the distinct expression pattern, determines lineage specification. MATERIALS AND METHODS: Using a retroviral vector, we introduced the GATA-3 gene into primary murine hematopoietic stem cells (HSC) and examined their development in in vitro suspension culture and colony-forming assays as well as in vivo competitive repopulation studies. RESULTS: Although GATA-3 expression normally is restricted to lymphoid precursor and committed T cells, overexpression of GATA-3 in HSC results in cessation of cell expansion followed by selective induction of megakaryocytic and erythroid differentiation and inhibition of myeloid and lymphoid precursor development in liquid suspension culture and in vitro colony-forming assays. Competitive repopulation studies show that transplanted GATA-3-expressing HSC/progenitor cells give one wave of erythrocyte development but fail to expand in the bone marrow or to reconstitute other lineages. CONCLUSIONS: The selective megakaryocytic/erythroid differentiation in HSC with enforced GATA-3 expression suggests a functional redundancy among GATA proteins and indicates that the specific lineage fate determination by individual GATA proteins is largely regulated at the level of expression in a lineage and developmental-stage restricted fashion, whereas the identity of the GATA factor may not be as important. PMID- 11495704 TI - Monoclonal antibody BB9 raised against bone marrow stromal cells identifies a cell-surface glycoprotein expressed by primitive human hemopoietic progenitors. AB - OBJECTIVE: The identification of cell-surface antigens whose expression is limited to primitive hematopoietic progenitor cells (HPC) is of major value in the identification, isolation, and characterization of candidate stem cells in human hemopoietic tissues. Based on the observation that bone marrow stromal cells and primitive HPC share several cell-surface antigens, we sought to generate monoclonal antibodies to HPC by immunization with cultured human stromal cells. METHODS: BALB/c mouse were immunized with human bone marrow (BM)-derived stromal cells. Splenocytes isolated from immunized mice were fused with the NS-1 murine myeloma cell line and resulting hybridomas selected in HAT medium, then screened for reactivity against stromal cells, peripheral blood (PB), and BM cells. RESULTS: A monoclonal antibody (MAb), BB9, was identified based on its binding to stromal cells, a minor subpopulation of mononuclear cells in adult human BM, and corresponding lack of reactivity with leukocytes in PB. BB9 bound to a minor subpopulation of BM CD34(+) cells characterized by high-level CD34 antigen and Thy-1 expression, low-absent expression of CD38, low retention of Rhodamine 123, and quiescent cycle status as evidenced by lack of labeling with Ki67. CD34(+)BB9(+) cells, in contrast to CD34(+)BB9(-) cells, demonstrated a capacity to sustain hematopoiesis in pre-CFU culture stimulated by the combination of IL-3, IL-6, G-CSF, and SCF. BB9 also demonstrated binding to CD34(+) cells from mobilized PB. CONCLUSION: Collectively, these data therefore demonstrate that MAb BB9 identifies an antigen, which is selectively expressed by hierarchically primitive human HPC and also by stromal cells. PMID- 11495705 TI - alpha(v)beta(3) integrin engagement modulates cell adhesion, proliferation, and protease secretion in human lymphoid tumor cells. AB - OBJECTIVE: The mechanisms used by human lymphoproliferative diseases to invade locally and metastasize are thought to be similar to those developed by solid tumors, including cell proliferation and secretion of extracellular matrix degrading enzymes following adhesion to extracellular matrix proteins. Hence, the ability of Namalwa (Burkitt's lymphoma), U266 (multiple myeloma), and CEM (T-cell lymphoblastic leukemia) cells to interact with the extracellular matrix components vitronectin and fibronectin was determined. Fresh bone marrow plasma cells from patients with multiple myeloma also were studied. MATERIALS AND METHODS: Engagement of alpha(v)beta(3) integrin, formation and protein composition of focal adhesion contacts on the cell surface, phosphorylation of several signal transduction proteins in the contacts, cell proliferation, and enzyme secretion were studied following adhesion to vitronectin and fibronectin. RESULTS: All three lines adhered to immobilized vitronectin and fibronectin. Adhesion was fully prevented by neutralizing monoclonal anti-alpha(v)beta(3) integrin antibody. Integrin engagement caused the formation of phosphorylated pp60(src)/focal adhesion kinase complexes and the aggregation of focal adhesion plaques containing the beta(3) integrin subunit, the cytoskeletal proteins vinculin, cortactin, and paxillin, the tyrosine kinases focal adhesion kinase and pp60(src), the adapter protein Grb-2, and the mitogen-activated protein kinase ERK-2. Free and immobilized vitronectin and fibronectin stimulated the proliferation of cells under serum-free conditions and the production and release of urokinase-type plasminogen activator, and increased the release of the activated forms of matrix metalloproteinase-2 and matrix metalloproteinase-9 in an alpha(v)beta(3) integrin-dependent manner. Similar results were obtained in myeloma plasma cells. CONCLUSIONS: The demonstrated ability of lymphoid tumor cells to interact with the extracellular matrix components vitronectin and fibronectin via alpha(v)beta(3) integrin can be interpreted as evidence of a novel mechanism for their invasion and spreading. This interaction allows them to adhere to the substratum and enhances their proliferation and protease secretion. PMID- 11495706 TI - Circulating natural IgM antibodies and their corresponding human cord blood cell derived Mabs specifically combat the Tat protein of HIV. AB - OBJECTIVE: IgM antibodies reactive with each of two specifically defined sequences of HIV Tat protein have been identified in sera from both HIV(+) and normal (HIV(-)) humans. This study was designed to confirm that those antibodies are innate immune factors capable of restriction of specific mechanisms of HIV pathogenicity attributed to the Tat protein. MATERIALS AND METHODS: Antibody secreting hybridomas were generated from human cord blood cells and processed for monoclonality. Those Mabs reactive with each of the sequences of Tat with which the circulating antibodies are reactive were isolated and their heavy and light chains identified and DNA sequenced. Pools of IgM isolated from blood of normal humans, chimpanzees, rhesus macaques, and mice and the isolated Tat reactive Mabs were tested for capacity to inhibit Tat-induced human T-cell apoptosis. RESULTS: Human and chimpanzee IgM pools, as well as the human cord blood cell-derived Mabs, showed a definite capacity to inhibit the Tat-induced apoptosis, while the IgM pools of rhesus macaques or of mice did not. CONCLUSIONS: These studies establish that the circulating IgM of normal humans include innate antibodies capable of restriction of HIV Tat-induced pathogenesis. That capacity is shared by chimpanzee IgM but not by IgM of other primates or of mice. The identification of those human circulating antibodies as innate is confirmed by the display of similar epitopic identity and apoptosis inhibition capacity by Mabs from human cord blood cell hybridomas. Thus, the arsenal of human cord blood cell hybridomas provides a resource by which, specifically, the potential therapeutic role of the identified HIV Tat-reactive Mabs and, broadly, the fundamental role of innate antibodies in infection control may be explored. PMID- 11495707 TI - Augmentation of megakaryocytopoiesis within the hematopoietic microenvironment of human granulocyte colony-stimulating factor transgenic mice. AB - OBJECTIVE: Megakaryocytopoiesis was dramatically augmented in human granulocyte colony-stimulating factor transgenic mice (G-Tg) compared to littermates. We examined the characteristics of megakaryocytes and megakaryocyte progenitor cells in these mice. MATERIALS AND METHODS: The numbers of colony-forming unit megakaryocytes (CFU-MK) and megakaryocytes in hematopoietic organs were counted. The megakaryocytes of G-Tg were examined ultrastructurally, and bone marrow transplantation studies using congenic G-Tg (Ly5.2) and C57BL/6 (Ly5.1) were performed. The number of day-14 colony-forming unit spleen (CFU-S) that contained megakaryocytes in [Ly5.1 > G-Tg] and [G-Tg > Ly5.1] mice also was counted. RESULTS: The number of CFU-MK increased markedly in the spleen, bone marrow, and peripheral blood. The number of megakaryocytes in the spleen and bone marrow also were increased in G-Tg mice. Ultrastructural analyses revealed that megakaryocytes in G-Tg mice were immature. Bone marrow transplantation studies of [Ly5.1 > G-Tg] mice resulted in a significantly increased number of megakaryocytes compared to [G-Tg > Ly5.1] mice. The number of day-14 CFU-S that contained megakaryocytes was increased markedly in [Ly5.1 > G-Tg] mice compared to [G-Tg > Ly5.1] mice. In vitro differentiation of megakaryocytes in [Ly5.1 > G Tg] mice was induced by interleukin-11 and thrombopoietin. CONCLUSION: The results showed that the hematopoietic marrow microenvironment of G-Tg is important in augmenting megakaryocytopoiesis. [Ly5.1 > G-Tg] mice are potentially useful as a source of murine megakaryocytes and their progenitors. PMID- 11495708 TI - Multipotent hematopoietic progenitor cells immortalized by Lhx2 self-renew by a cell nonautonomous mechanism. AB - OBJECTIVE: Direct molecular and cellular studies of hematopoietic stem cells (HSCs) are hampered by the low levels of HSCs in hematopoietic tissues. To address these issues, we generated immortalized multipotent hematopoietic precursor cell (HPC) lines by expressing the LIM-homeobox gene Lhx2 (previously LH2) in hematopoietic progenitors derived from embryonic stem cells differentiated in vitro. MATERIALS AND METHODS: To validate further the relevance of the HPC lines as a model for normal HSCs, we analyzed in detail the growth requirements of HPC lines in vitro. RESULTS: Lhx2 immortalized the HPC lines by a putatively novel and cell nonautonomous mechanism. Self-renewal of the HPC lines is dependent on functional Lhx2 expression. Most early-acting hematopoiesis related growth factors show synergistic effects on the HPC lines, whereas late acting factors do not induce differentiation by themselves. Transforming growth factor-beta(1) is a potent inhibitor of proliferation of the HPC lines. HPC lines form cobblestone areas with high efficiency when seeded onto stromal cell lines, and the cobblestone area-forming cell can be maintained in these cultures for several months. CONCLUSIONS: Our data show that, in many respects, HPC lines are similar to normal hematopoietic progenitor/stem cells on the cellular level, in contrast to most previously described multipotent hematopoietic cell lines. The cell nonautonomous mechanism for immortalization of the HPC lines suggests that Lhx2 regulates, directly or indirectly, soluble mediators involved in self renewal of the HPC lines. PMID- 11495709 TI - CD34(+) cord blood cells expressing cutaneous lymphocyte-associated antigen are enriched in granulocyte-macrophage progenitors and support extensive amplification of dendritic cell progenitors. AB - OBJECTIVE: We evaluated the frequency of hematopoietic progenitor cells (HPC) in CD34(+)CLA(+) (cutaneous lymphocyte-associated antigen) and CD34(+)CLA(-) cord blood cells, and followed cellular growth and HPC production during cultures in Flt3 ligand, thrombopoietin, and stem cell factor (FTS). MATERIALS AND METHODS: Immunomagnetic bead-purified CD34(+) cells were sorted into CD34(+)CLA(+) or CD34(+)CLA(-) cells. HPC frequency was assessed by clonal assays in methylcellulose either ex vivo or after, 7, 14, or 21 days of culture with FTS. Dendritic cell (DC) progenitors were evaluated after induction of FTS-amplified cells into DC using secondary cultures containing granulocyte-macrophage colony stimulating factor and interleukin-4. RESULTS: Ex vivo, granulocyte-macrophage progenitors were more frequent and erythroid progenitors were less frequent in the CLA(+) fraction. In FTS culture, CD34(+)CLA(+) cells produced greater absolute numbers of CD34(+) cells, granulocyte-macrophage-, erythroid-, and DC (including Langerhans cell-related) progenitors compared to CD34(+)CLA(-) cells. In CD34(+)CLA(+) cultures, CLA(+) cells steadily decreased with time, and CD34(+)CLA(-) cells appeared. In CD34(+)CLA(-) cultures, CLA(+) cells were generated, increased up to day 7, and decreased thereafter. CLA was expressed only on CD34(-) cells in these cultures. Ex vivo, CD34(+)CLA(+) cells could be subdivided further into CD38(low) and CD38(high) cells. Cord blood and growth factor-mobilized CD34(+) cells contained more CLA(+)CD38(low) cells than nonmobilized peripheral blood CD34(+) cells and proliferated more extensively with FTS than the latter cells. CONCLUSIONS: CD34(+)CLA(+) cells contain a rather immature progenitor capable of high proliferation and extensive amplification of HPC in vitro. This progenitor may be localized in the CD34(+)CLA(+)CD38(low) fraction. In addition, cultures of CD34(+)CLA(+) cells from cord blood produced CD34(+)CLA(-) cells, suggesting that these cells may derive directly from CD34(+)CLA(+) cells in vivo. PMID- 11495710 TI - A preliminary study of the utility of the brief hospice inventory. AB - The increasing number of palliative care patients necessitates a simple, reliable instrument to routinely measure outcomes among hospice patients. We tested the utility of the Brief Hospice Inventory (BHI) to assess outcomes of hospice patients and estimations of patients' outcomes by nurse caregivers. In a prospective study, 145 home-based hospice patients were enrolled in the study from VistaCare Hospice. During the first week of admission, patients and nurse caregivers completed the BHI, which assessed patients' symptoms, satisfaction with care, and quality of life. Factor analysis supported a two-factor structure for the BHI for patients and caregivers, including a symptom subscale and quality of life subscale. Patients with severe symptoms showed improvement on the symptom subscale, but not the quality of life subscale, during the first 2 weeks after admission. The BHI shows utility in measuring hospice patients' symptom severity and quality of life over time. PMID- 11495711 TI - Prospective assessment of symptom palliation for patients attending a rapid response radiotherapy program. feasibility of telephone follow-up. AB - Clinical trials generally include motivated patients with relatively good performance status. This can result in an overestimation of the effectiveness of an intervention. Clinic follow-up protocols for outcome assessment after palliative treatments suffer from high attrition rates. In this study, the feasibility of telephone follow-up for the assessment of symptom palliation in patients receiving outpatient palliative radiotherapy as a tool to evaluate outcome was examined. Patients referred for palliative radiotherapy were asked to rate their symptom distress using the modified Edmonton Symptom Assessment System (ESAS) at initial consultation. Patient demographics and analgesic consumption were collected. For those who received radiotherapy, follow-up was conducted through telephone interviews at week 1, 2, 4, 8, and 12 post-treatment using the same modified ESAS and analgesic diary. One hundred ninety patients received radiotherapy to 256 sites from January to August 1999. Seventy-eight patients (41%) died during the 12-week follow-up period. The percentage of surviving patients responding to the telephone interview ranged from 63% to 68% during the 12-week study. Telephone follow-up is a feasible tool for the prospective outcome assessment of symptom palliation in this population. It compares well to clinic visits or mailed questionnaires. However, to improve the follow-up rates, other modalities may also need to be implemented. PMID- 11495712 TI - Uncertainty and opposition of medical students toward assisted death practices. AB - To explore medical students' views of assisted death practices in patient cases that describe different degrees and types of physical and mental suffering, an anonymous survey was administered to all students at one medical school. Respondents were asked about the acceptability of assisted death activities in five patient vignettes and withdrawal of life support in a sixth vignette. In the vignettes, actions were performed by four possible agents: the medical student personally; a referral physician; physicians in general; or non-physicians. Of 306 medical students, 166 (54%) participated. Respondents expressed opposition or uncertainty about assisted death practices in the five patient cases that illustrated severe forms of suffering which were secondary to amyotrophic lateral sclerosis, treatment-resistant depressive and somatoform disorders, antisocial and sexually violent behavior, or AIDS. Students supported the withdrawal of life support in the sixth vignette depicting exceptional futility secondary to AIDS. Students were especially opposed to their own involvement and to the participation of non-physicians in assisted death activities. Differences in views related to sex, religious beliefs, and personal philosophy were found. Medical students do not embrace assisted death practices, although they exhibit tolerance regarding the choices of medical colleagues. How these attributes of medical students will translate into future behaviors toward patients and peers remains uncertain. Medical educators must strive to understand the perspectives of physicians-in-training. Expanded, empirically informed education that is attuned to the attitudes of medical students may be helpful in fulfilling the responsibility of imparting optimal clinical care skills. PMID- 11495713 TI - Low-concentration morphine infusion does not compromise packed red blood cell transfusion. AB - Regulations currently prohibit co-administration (through the same line) of red blood cell transfusions with continuous morphine infusions for pain management, resulting in additional intravenous access or interrupted analgesic therapy in seriously ill children. Packed cells that had been in contact with morphine 0.1 or 1.0 mg/mL and infused through a mock central venous catheter system showed no evidence of hemolysis when compared with control samples. There is thus no need to interrupt analgesic therapy or start another venous access line in order to give a coincident blood transfusion. PMID- 11495714 TI - Equianalgesic dose ratios for opioids. a critical review and proposals for long term dosing. AB - Clinicians involved in the opioid pharmacotherapy of cancer-related pain should be acquainted with a variety of opioids and be skilled in the selection of doses when the type of opioid or route of administration needs changing. The optimal dose should avoid under-dosing or overdosing, both associated with negative outcomes for the patient. Although equianalgesic dose tables are generally used to determine the new doses in these circumstances, the evidence to support the ratios indicated in these tables largely refers to the context of single dose administration. The applicability of these ratios to the setting of chronic opioid administration has been questioned. A systematic search of published literature from 1966 to September 1999 was conducted to critically appraise the emerging evidence on equianalgesic dose ratios derived from studies of chronic opioid administration. There were six major findings: 1) there exists a general paucity of data related to long-term dosing and studies are heterogeneous in nature; 2) the ratios exhibit extremely wide ranges; 3) methadone is more potent than previously appreciated; 4) the ratios related to methadone are highly correlated with the dose of the previous opioid; 5) the ratio may change according to the direction the opioid switch; and 6) discrepancies exist with respect to both oxycodone and fentanyl. Overall, these findings have important clinical implications for clinicians and warrant consideration in the potential revision of current tables. The complexity of the clinical context in which many switches occur must be recognized and also appreciated in the design of future studies. PMID- 11495715 TI - Economic evaluation of multidisciplinary pain management in chronic pain patients: a qualitative systematic review. AB - In this qualitative systematic review, we have evaluated studies of the economic effectiveness of multidisciplinary pain treatment in chronic non-malignant pain patients. Published reports were identified from a systematic search of bibliographic databases (MEDLINE and EMBASE) and reference lists of retrieved reports. Fourteen reports of nine studies of patients suffering from back pain, fibromyalgia, and mixed chronic pain conditions were considered to be appropriate as economic analyses. In the selected studies, we found serious methodological problems in study designs and application of outcome measures. The quality of the cost measurements was characterized by an apparent lack of tradition using economic methodology. This review does not give an answer to whether multidisciplinary pain management in chronic pain patients is cost-effective or not. Application of standard methods of costing and outcome measurement are essential before studies of cost-effectiveness in multidisciplinary pain treatment can be used in decision-making and planning. PMID- 11495716 TI - Diphenhydramine as an analgesic adjuvant in refractory cancer pain. AB - Clinical and animal data suggest that antihistamines may have efficacy in the management of pain. While many mechanisms of action have been proposed for the analgesic action of antihistamines, the exact mechanism is unknown. Controlled clinical trials in different pain models have demonstrated that antihistamines have direct and adjuvant analgesic activity. We report three patients with advanced cancer pain refractory to adjuvants and oral, intravenous, and epidural opioids, who achieved sustained pain relief after the repeated administration of diphenhydramine. Diphenhydramine may be useful in the treatment of neuropathic and nociceptive pain that has failed to respond to treatment with opioids and adjuvant analgesics. We suggest a starting dose of 25 mg of oral or parenteral diphenhydramine every 6 to 8 hours, with titration to effect. PMID- 11495717 TI - Olanzapine for the treatment of fibromyalgia symptoms. AB - Fibromyalgia is a chronic condition that is diagnosed primarily by the presence of generalized pain along with tenderness on palpation of certain body regions. Unfortunately, the pharmacological treatment of fibromyalgia remains problematic. Two patients are described who highlight the use of the atypical neuroleptic olanzapine for the control of symptoms related to fibromyalgia. Prior to the use of olanzapine, both patients had received a multitude of treatments, none of which greatly improved their ability to function in daily activities. With olanzapine, both patients reported a significant decrease in pain and marked improvement in daily functioning. In one case, the pain returned during a period of time when olanzapine was discontinued, an effect that was reversed when olanzapine was reintroduced. The paucity of serious side effects (i.e., extrapyramidal signs) with the atypical neuroleptic olanzapine strongly favors further exploration and use of this drug for the treatment of fibromyalgia symptoms. PMID- 11495718 TI - Signal transduction via the growth hormone receptor. AB - Rapid progress has been made recently in the definition of growth hormone (GH) receptor signal transduction pathways. It is now apparent that many cytokines, including GH, share identical or similar signalling components to exert their cellular effects. This review provides a brief discourse on the signal transduction pathways, which have been demonstrated to be utilized by GH. The identification of such pathways provides a basis for understanding the pleiotropic actions of GH. The mechanisms by which the specific cellular effects of GH are achieved remain to be elucidated. PMID- 11495719 TI - Fas-mediated apoptosis in a rat acinar cell line is dependent on caspase-1 activity. AB - Apoptosis plays an important role in the dysfunction of exocrine glands. Fas is a death-inducing receptor found on many types of cells including epithelial acinar cells. To elucidate the intracellular mechanism of Fas-mediated cell death in exocrine glands, an epithelial acinar cell line, SMG-C6, was studied. Caspase-1, 3, -8, and -9 activities were elevated in SMG-C6 cells after the induction of apoptosis by soluble Fas ligand (FasL). The activation of caspase-1 and -8 occurred prior to caspase-3 and -9 activation. The caspase-1 inhibitor, zYVAD fmk, was effective in preventing cell death, whereas the caspase-3 and -8 inhibitors (ac-DEVD-CHO and ac-IETD-CHO, respectively) were not. zYVAD-fmk was able to inhibit caspase-3 activation indicating that caspase-1 is upstream to caspase-3. Furthermore, kinetic studies show that caspase-1 is an early event in the Fas apoptotic pathway. This study shows that caspase-1 participates in Fas mediated apoptosis of epithelial cells by initiating the caspase cascade. PMID- 11495720 TI - The interaction of dystrophin with beta-dystroglycan is regulated by tyrosine phosphorylation. AB - Dystrophin and the dystrophin-associated protein complex (DAPC) have recently been implicated in cell signalling events. These proteins are ideally placed to transduce signals from the extracellular matrix (ECM) to the cytoskeleton. Here we show that beta-dystroglycan is tyrosine-phosphorylated in C2/C4 mouse myotubes. Tyrosine phosphorylation was detected by mobility shifts on SDS polyacrylamide gels (SDS-PAGE) and confirmed by immunoprecipitation and two dimensional gel electrophoresis. The potential functional significance of this tyrosine phosphorylation was investigated using peptide 'SPOTs' assays. Phosphorylation of tyrosine in the 15 most C-terminal amino acids of beta dystroglycan disrupts its interaction with dystrophin. The tyrosine residue in beta-dystroglycan's WW-binding motif PPPY appears to be the most crucial in disrupting the beta-dystroglycan-dystrophin interaction. beta-dystroglycan forms the essential link between dystrophin and the rest of the DAPC. This regulation by tyrosine phosphorylation may have implications in the pathogenesis and treatment of Duchenne's muscular dystrophy (DMD). PMID- 11495721 TI - Tumour necrosis factor-alpha enhances bradykinin-induced signal transduction via activation of Ras/Raf/MEK/MAPK in canine tracheal smooth muscle cells. AB - Inhalation of tumour necrosis factor-alpha (TNF-alpha) induced a bronchial hyperreactivity to contractile agonists. However, the mechanisms of TNF-alpha involved in the pathogenesis of bronchial hyperreactivity were not completely understood. Therefore, we investigated the effect of TNF-alpha on bradykinin (BK) induced inositol phosphate (IP) accumulation and Ca(2+) mobilization, and up regulation of BK receptor density in canine cultured tracheal smooth muscle cells (TSMCs). Pretreatment of TSMCs with TNF-alpha potentiated BK-induced IP accumulation and Ca(2+) mobilization. However, there was no effect on the IP response induced by endothelin-1 (ET-1), 5-hydroxytryptamine (5-HT), and carbachol. Pretreatment with PDGF B-chain homodimer (PDGF-BB) also enhanced BK induced IP response. These enhancements induced by TNF-alpha and PDGF-BB might be due to an increase in BK B(2) receptor density (B(max)), since [3H]BK binding to TSMCs was inhibited by the B(2) selective agonist and antagonist, BK and Hoe 140, but not by the B(1) selective reagents. The enhancing effects of TNF-alpha and PDGF-BB were attenuated by PD98059 (an inhibitor of activation of MAPK kinase, MEK) and cycloheximide (an inhibitor of protein synthesis), suggesting that TNF alpha may share a common signalling pathway with PDGF-BB via protein(s) synthesis in TSMCs. Furthermore, overexpression of dominant negative mutants, H-Ras-15A and Raf-N4, significantly suppressed p42/p44 mitogen-activated protein kinase (MAPK) activation induced by TNF-alpha and PDGF-BB and attenuated the effect of TNF alpha on BK-induced IP response, indicating that Ras and Raf may be required for activation of these kinases. These results suggest that the augmentation of BK induced responses produced by TNF-alpha might be, at least in part, mediated through activation of Ras/Raf/MEK/MAPK pathway in TSMCs. PMID- 11495722 TI - cAMP potentiates H(2)O(2)-induced ERK1/2 phosphorylation without the requirement for MEK1/2 phosphorylation. AB - In Jurkat T lymphocytes, hydrogen peroxide (H(2)O(2)) potentiates the phosphorylation level of extracellular signal-regulated kinase 1 and 2 (ERK1/2) caused by T cell receptor (TCR) stimulation with anti-CD3 and anti-CD28 or anti CD3 alone. Submillimolar concentrations of H(2)O(2)-induced phosphorylation of ERK1/2 and MAP/ERK kinase 1 and 2 (MEK1/2) without antigenic stimulation. H(2)O(2) also induced the electrophoretic mobility shift of Lck from 56 to 60 kDa. The MEK inhibitor, PD98059 attenuated ERK1/2 and MEK1/2 phosphorylation, as well as the migration shift of Lck induced by H(2)O(2). The phospholipase C (PLC) inhibitor, U73122, and EGTA reduced the phosphorylation of both ERK1/2 and MEK1/2 induced by H(2)O(2). Interestingly, an increase of intracellular cAMP level with forskolin or 8-(4-chlorophenylthio)-cAMP augmented ERK1/2 phosphorylation by H(2)O(2), while inhibiting MEK1/2 phosphorylation by H(2)O(2). These results demonstrate an alternative pathway that results in augmentation of ERK1/2 phosphorylation without concomitant MEK1/2 phosphorylation in T cells. PMID- 11495723 TI - MEK, ERK, and p90RSK are present on mitotic tubulin in Swiss 3T3 cells: a role for the MAP kinase pathway in regulating mitotic exit. AB - The mitogen-activated protein (MAP) kinase pathway has been implicated in cell cycle control for some time. Several reports have suggested a role for this pathway in growth factor stimulation of DNA synthesis, while other reports have proposed a role in the transition of cells through mitosis. Here, we have examined the potential involvement of the extracellular signal-related kinase (ERK)1/2 MAP kinases, their upstream regulators, and downstream effectors in the regulation of mitosis. Inhibition of MAP kinase/ERK kinase (MEK) activity reduced the serum-stimulated DNA synthesis and proliferation of Swiss 3T3 cells. To study the potential mechanisms of this effect, we examined the subcellular localization of members of the MAP kinase pathway including regulators (MEK1/2), substrates (90-kDa ribosomal S6 kinases (RSKs): RSK1, RSK2 and RSK3), and ERK itself. We show that there is enrichment of ERK, MEK, and the RSK enzymes on both the spindle and midbody tubulin of dividing cells. Inhibition of MEK1/2 activity in cells released from mitotic arrest results in an inability of cells to complete mitosis. This failure to exit mitosis correlated with altered cyclin-dependent kinase (cdk) activities. Thus, the MAP kinase pathway may act to coordinate passage through mitosis in Swiss 3T3 fibroblasts by regulation of cdk activity. PMID- 11495724 TI - Insulin and heregulin-beta1 upregulate guanylyl cyclase C expression in rat hepatocytes: reversal by phosphodiesterase-3 inhibition. AB - Guanylyl cyclase C (GC-C) is the receptor for the hormones guanylin and uroguanylin. Although primarily expressed in the rat intestine, GC-C is also expressed in the liver during neonatal or regenerative growth or during the acute phase response. Little is known about the hepatic regulation of GC-C expression. The influence of various hepatic growth or acute phase regulators on GC-C expression was evaluated by immunoblot analysis of protein from primary rat hepatocytes grown in a serum-free medium. Insulin and heregulin-beta1 strongly stimulated GC-C expression by 24 h of cell culture. Several different hormones and agents suppressed this action, including transforming growth factor beta (TGF beta), as well as inhibitors of phosphatidylinositol 3-kinase (PI-3-kinase) and phosphodiesterase 3 (PDE-3, an insulin- and PI-3-kinase-dependent enzyme). The compartmental downregulation of cAMP levels by PDE-3 may be a critical step in the hormonal action that culminates in GC-C synthesis. PMID- 11495725 TI - Activation of the transcription factor NF-kappaB by the erythropoietin receptor: structural requirements and biological significance. AB - The transcription factor nuclear factor kappa B (NF-kappaB) has been implicated in the regulation of genes mainly involved in inflammation and immune response. We analysed the role of NF-kappaB in signalling pathways induced by the hematopoietic growth factor erythropoietin (EPO). Our data, obtained by electrophoretic mobility shift assays (EMSA) and reporter gene assays, show that the intracellular domain of the EPO receptor (EPOR) transmits signals leading to the activation of NF-kappaB. Studies employing an inhibitor specific for the EPOR associated tyrosine kinase JAK2 suggest that JAK2-dependent pathways are not involved. The induction of an NF-kappaB-triggered reporter gene construct was inhibited by cotransfection of dominant negative forms of the src kinase Lyn, but not by dominant negative JAK2. Using epidermal growth factor (EGF)/EPOR hybrids containing mutant forms of the EPOR intracellular domain, we were able to further define the critical structures for the induction of NF-kappaB. The data show that although the activity of JAK2 seems to be dispensable, its association to the receptor, as well as the phosphorylation of membrane proximal tyrosine residues, are essential. Furthermore, the functional analysis of different receptor forms revealed a correlation of the abilities to induce NF-kappaB activity and to generate antiapoptotic signals. PMID- 11495727 TI - Membranes. Engineering and design. PMID- 11495728 TI - Lipids lost, lipids regained. PMID- 11495729 TI - Stabilizing membrane proteins. AB - Membrane proteins can be extremely stable in a bilayer environment, but are often unstable and rapidly lose activity after detergent solubilization. Poor stability can preclude the detailed characterization of many membrane proteins. One way to alleviate this problem is to find more stable mutants of a membrane protein of interest. This approach is made tractable by the finding that stability-enhancing mutations appear to be relatively common in membrane proteins. PMID- 11495730 TI - Channel-tunnels. AB - TolC and its many homologues comprise an alpha-helical transperiplasmic tunnel embedded in the bacterial outer membrane by a contiguous beta-barrel channel, providing a large exit duct for diverse substrates. The 'channel-tunnel' is closed at its periplasmic entrance, but can be opened by an 'iris-like' mechanism when recruited by substrate-engaged proteins in the cytosolic membrane. PMID- 11495731 TI - Potassium channels: life in the post-structural world. AB - More than three years have passed since the first structure of a potassium channel protein revealed fundamental molecular details of a platform for ion selective conduction. Recent efforts have turned to understanding what this structure tells us about potassium channel structure and function in general and, most importantly, which questions remain unanswered. Successes in solving membrane protein structures are still hard won and slow. High-resolution studies of cytoplasmic channel domains and channel-associated proteins, the most tractable entry points for dissecting large, complex eukaryotic channels, are revealing a modularity of function commonly seen in many other biological systems. Studies of these domains bring into sharp focus issues of channel regulation, how these domains and associated proteins are coupled to the transmembrane domains to influence channel function, and how ion channels are integrated into cellular signaling pathways. PMID- 11495732 TI - Bacteriorhodopsin. AB - High-resolution maps from X-ray diffraction of bacteriorhodopsin and some of its photointermediates have yielded insights into how the isomerization of the bound retinal drives ion transport. Although important mechanistic details are still undecided, the events of the photochemical cycle are now understood to reflect changes in specific hydrogen bonds of protein groups and bound water molecules in response to motions of the retinal chain. PMID- 11495733 TI - Crystal structure of rhodopsin: implications for vision and beyond. AB - A heptahelical transmembrane bundle is a common structural feature of G-protein coupled receptors (GPCRs) and bacterial retinal-binding proteins, two functionally distinct groups of membrane proteins. Rhodopsin, a photoreceptor protein involved in photopic (rod) vision, is a prototypical GPCR that contains 11-cis-retinal as its intrinsic chromophore ligand. Therefore, uniquely, rhodopsin is a GPCR and also a retinal-binding protein, but is not found in bacteria. Rhodopsin functions as a typical GPCR in processes that are triggered by light and photoisomerization of its ligand. Bacteriorhodopsin is a light driven proton pump with an all-trans-retinal chromophore that photoisomerizes to 13-cis-retinal. The recent crystal structure determination of bovine rhodopsin revealed a structure that is not similar to previously established bacteriorhodopsin structures. Both groups of proteins have a heptahelical transmembrane bundle structure, but the helices are arranged differently. The activation of rhodopsin involves rapid cis-trans photoisomerization of the chromophore, followed by slower and incompletely defined structural rearrangements. For rhodopsin and related receptors, a common mechanism is predicted for the formation of an active state intermediate that is capable of interacting with G proteins. PMID- 11495734 TI - High-resolution structures and dynamics of membrane protein--lipid complexes: a critique. AB - Crystallographic analysis of lipidic components in complex with membrane proteins reveals molecules exhibiting well-ordered polar or hydrophobic moieties in contact with protein. As dynamic methods indicate high exchange rates, the interpretation of structural data requires a detailed knowledge of the specificity, affinity and cooperativity of lipid binding. PMID- 11495735 TI - Force as a probe of membrane protein structure and function. AB - Force measurement techniques are being used increasingly to explore the mechanical properties of proteins, as well as the structural origins of intermolecular forces. Developments in instrumentation and the increasing availability of engineered and purified membrane proteins have widely expanded the range of biological systems that can be addressed. Within the past year, force measurements have identified novel mechanisms of binding between cell surface proteins, as well as the mechanical properties of integral membrane proteins and the intramolecular interactions that stabilize their structures. PMID- 11495736 TI - Structures of lipid-DNA complexes: supramolecular assembly and gene delivery. AB - Recently, there has been a flurry of experimental work on understanding the supramolecular assemblies that are formed when cationic liposomes (CLs) are mixed with DNA. From a biomedical point of view, CLs (vesicles) are empirically known to be carriers of genes (sections of DNA) in nonviral gene delivery applications. Although viral-based carriers of DNA are presently the most common method of gene delivery, nonviral synthetic methods are rapidly emerging as alternative carriers, because of their ease of production and nonimmunogenicity (viral carriers very often evoke an undesirable and potentially lethal immune response). At the moment, cationic-lipid-based carriers have emerged as the most popular nonviral method to deliver genes in therapeutic applications, for example, CL carriers are used extensively in clinical trials worldwide. However, because the mechanism of transfection (the transfer of DNA into cells by CL carriers, followed by expression) of CL--DNA complexes remains largely unknown, the measured efficiencies are, at present, very low. The low transfection efficiencies of current nonviral gene delivery methods are the result of poorly understood transfection-related mechanisms at the molecular and self-assembled levels. Recently, work has been carried out on determining the supramolecular structures of CL--DNA complexes by the quantitative technique of synchrotron X ray diffraction. When DNA is mixed with CLs (composed of mixtures of cationic DOTAP and neutral DOPC lipids), the resulting CL--DNA complex consists of a multilamellar structure (L(alpha)(C)) comprising DNA monolayers sandwiched between lipid bilayers. The existence of a different columnar inverted hexagonal (H(II)(C)) phase in CL--DNA complexes was also demonstrated using synchrotron X ray diffraction. Ongoing functional studies and optical imaging of cells are expected to clarify the relationship between the supramolecular structures of CL- DNA complexes and transfection efficiency. PMID- 11495739 TI - The design of self-replicating helical peptides. AB - The self-assembly of helical peptides and information transfer through autocatalysis and cross-catalysis are the foundation of peptide-based molecular evolution models. Many fundamental properties of living systems, such as environmental sensitivity, chiroselectivity, cross-catalysis, dynamic error correction and conditional selection, are exhibited by various self-replicating peptide systems. Recently, advances have been made in the design of peptide systems with autocatalytic and cross-catalytic properties. PMID- 11495738 TI - The design of antiparallel coiled coils. AB - Recent structural studies have highlighted the importance of antiparallel coiled coils in nature. In addition, well-behaved, model antiparallel coiled coils have been designed and used for the reassembly of protein fragments and for the study of the energetic contributions of various interactions to helix orientation specificity. Finally, high-resolution structural data are available for designed helical bundles, allowing an evaluation of the success of state-of-the-art protein design efforts. PMID- 11495740 TI - Core-directed protein design. AB - For various reasons, it seems sensible to redesign or design proteins from the inside out. Past approaches in this field have involved iterations of mutagenesis and characterisation to 'evolve' designs. Increasingly, combinatorial approaches are being taken to select 'fit' sequences from libraries of variant proteins. In particular, in silico methods have been used to good effect. More recently, experimental methods have been developed and improved. We are now in a position to redesign stability and function into natural protein frameworks confidently and to attempt de novo designs for more ambitious targets. PMID- 11495741 TI - Exploring protein interactions by interaction-induced folding of proteins from complementary peptide fragments. AB - The study of protein--protein interactions is central to understanding the chemical machinery that makes up the living cell. Until recently, facile methods to study these processes in intact, living cells have not existed. Furthermore, the assignment of function to novel proteins relies on demonstrating interactions of these proteins with proteins of known function. This review describes an experimental strategy, devised to study protein--protein interactions in any intact living cells based on protein-fragment complementation assays. Applications to quantitative analysis of interactions, allosteric processes and cDNA library screening are discussed. Recently, the feasibility of employing this strategy in genome-wide biochemical pathway mapping efforts has been demonstrated. PMID- 11495742 TI - Design of RNA-binding proteins and ligands. AB - The rapidly expanding database of RNA structures and protein complexes is beginning to lead to the successful design of specific RNA-binding molecules. Recent combinatorial and structure-based approaches have utilized known nucleic acid-binding scaffolds from both proteins and small molecules to display a relatively small set of functional groups often used in protein--RNA recognition. Several studies have shown that the tethering of multiple binding modules can enhance RNA-binding affinity and specificity, a strategy also commonly used in DNA recognition. PMID- 11495744 TI - Engineering and design in the bioelectrochemistry of metalloproteins. AB - Engineered metalloproteins offer interesting systems for electrochemical studies of protein structure/function and their applications in nanobiotechnology. Scanning probe microscopy and cyclic voltammetry of engineered metalloproteins and electrodes have proved to be a powerful combination of tools contributing to the field of bioelectrochemistry. The ability to engineer tags, such as histidine tags and biotin-acceptor peptides, and to site-specifically introduce cysteine residues enabled the creation of ordered immobilised protein structures that can be characterised both electrochemically and topographically. Gene fusion and de novo combinatorial synthesis of metalloproteins are emerging to provide structures with the desired electrochemical properties. PMID- 11495743 TI - Metalloprotein and redox protein design. AB - Metalloprotein and redox protein design are rapidly advancing toward the chemical synthesis of novel proteins that have predictable structures and functions. Current data demonstrate a breadth of successful approaches to metallopeptide and metalloprotein design based on de novo, rational and combinatorial strategies. These sophisticated synthetic analogs of natural proteins constructively test our comprehension of metalloprotein structure/function relationships. Additionally, designed redox proteins provide novel constructs for examining the thermodynamics and kinetics of biological electron transfer. PMID- 11495745 TI - Chemoinformatics: manipulating chemical information to facilitate decision-making in drug discovery. PMID- 11495746 TI - Could a vaccine treat spinal cord injury? PMID- 11495747 TI - New hope for sickle cell anaemia. PMID- 11495751 TI - 20 years on: Donald Francis discusses advances in the fight against HIV. PMID- 11495748 TI - Novel and topical: immune-response modifier for skin cancer. PMID- 11495752 TI - Is there a future for neural grafting? PMID- 11495753 TI - The broader applications of uHTS. PMID- 11495754 TI - Proteomics: delivering new routes to drug discovery - Part 2. PMID- 11495755 TI - High content screening - from cells to data to knowledge. PMID- 11495756 TI - Pharmacogenetics of the human drug-transporter gene MDR1: impact of polymorphisms on pharmacotherapy. AB - The blood- and tissue-concentrations, and thus the activity, of many drugs are influenced by factors that are subject to inter-individual variation. Variables that influence blood levels are metabolizing enzymes and transporters. Transporters control drug uptake, distribution and elimination. Transport by efflux pumps such as MDR1-encoded P-glycoprotein can influence the bioavailability of drugs. Knowledge of the transporter 'status' might allow for compensation of differences in drug uptake, such as by dose adjustment, which is important for drugs with narrow therapeutic windows. So far, intestinal expression of MDR1 has been determined by cumbersome methods, such as biopsies, although recently a functional polymorphism has been identified, which discriminates individual high or low-expressor alleles. As a result, clinical trials and therapy can be adapted to the 'MDR1-status' of individual patients. PMID- 11495757 TI - Pure compound libraries; a new perspective for natural product based drug discovery. AB - What often distinguishes the leaders in drug discovery and development from the rest is the quality of their compound libraries and the ease of access that they have to the information within those libraries. The screening of natural products can provide greater structural diversity than standard synthetic chemistry and offers significant opportunities for finding novel low molecular weight lead compounds. However, which strategy is the best for natural-product-based drug discovery? Two well established but relatively time consuming approaches are the screening of crude extracts and pre-fractionated extracts. This case study describes a third, pure-compound-screening approach, and discusses its benefits and pitfalls. PMID- 11495758 TI - Inhibition of osteopenia by low magnitude, high-frequency mechanical stimuli. AB - The identification of anabolic agents for the treatment of metabolic bone disease is a highly prized, and elusive, goal. In searching for the osteogenic (bone producing) constituents within mechanical stimuli, it was determined that high frequency (10-100 Hz) and low magnitude (<10 microstrain) stimuli were capable of augmenting bone mass and morphology, thereby benefiting both bone quantity and quality. Using animal models, it is shown that these mechanical signals can double bone-formation rates, inhibit disuse osteoporosis and increase the strength of trabecular bone by 25%. Considering that the magnitude of these mechanical signals are several orders of magnitude below those which cause damage to the bone tissue, it is proposed that this modality could be useful in the treatment of metabolic bone diseases. PMID- 11495759 TI - Anti-HIV activity of betulinic acid analogue YKFH312. PMID- 11495760 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 11495762 TI - A drug delivery system for the treatment of periodontitis. PMID- 11495761 TI - Combinatorial chemistry. PMID- 11495763 TI - Control of potato tuber sprouting. PMID- 11495764 TI - Ammonium toxicity and the real cost of transport. PMID- 11495765 TI - Using complex plant pedigrees to map valuable genes. PMID- 11495766 TI - A hydrophobin aids water-mediated dispersal of Cladosporium conidia. PMID- 11495767 TI - A hairy tale: Glabrous meets Werewolf. PMID- 11495768 TI - Luc-imaging the cold signal transduction pathway. PMID- 11495769 TI - Unravelling PTGS: SDE3 - an RNA helicase involved in RNA silencing in Arabidopsis. PMID- 11495787 TI - A quantitative model of the domain structure of the photosynthetic membrane. AB - A model is presented that gives a quantitative picture of the distribution of the photosynthetic components in the photosynthetic membrane of higher plants. A salient feature of the model is that most of the pigments are located in the grana where photosystem I and II carry out linear electron transport, whereas the stroma lamellae, which harbour <20% of the pigments, carry out photosystem-I mediated cyclic electron transport. This arrangement derives from the observation that more pigments are associated with photosystem I, which therefore captures more quanta than photosystem II. The excess pigments associated with photosystem I are thought to be located in the stroma lamellae. PMID- 11495788 TI - The U-box protein family in plants. AB - The U-box is a highly conserved domain recently identified at the C terminus of yeast UFD2, an E4 ubiquitination factor. In yeast, UFD2 is the only U-box containing protein, but there are two UFD2 homologs and several other proteins containing a U-box domain in humans. Intriguingly, a database search revealed 37 predicted proteins containing a U-box in Arabidopsis. The plant U-box (PUB) proteins form five distinct subclasses, suggesting that they play diverse roles. The ARC1 gene from Brassica, required for self-incompatibility, is currently the only PUB gene functionally characterized. Here, we discuss the characteristics and possible functions of the PUB gene family. PMID- 11495789 TI - When plant cells decide to divide. AB - Progression through the cell cycle is central to cell proliferation and fundamental to the growth and development of all multicellular organisms, including higher plants. The periodic activation of complexes containing cyclins and cyclin-dependent kinases mediates the temporal regulation of the cell-cycle transitions. Here, we highlight recent advances in the molecular controls of the cell cycle in plant cells, with special emphasis on how hormonal signals can modulate the regulation of cyclin-dependent kinases. PMID- 11495790 TI - Functional genomics of plant photosynthesis in the fast lane using Chlamydomonas reinhardtii. AB - Oxygenic photosynthesis by algae and plants supports much of life on Earth. Several model organisms are used to study this vital process, but the unicellular green alga Chlamydomonas reinhardtii offers significant advantages for the genetic dissection of photosynthesis. Recent experiments with Chlamydomonas have substantially advanced our understanding of several aspects of photosynthesis, including chloroplast biogenesis, structure-function relationships in photosynthetic complexes, and environmental regulation. Chlamydomonas is therefore the organism of choice for elucidating detailed functions of the hundreds of genes involved in plant photosynthesis. PMID- 11495791 TI - Signal transmission in the plant immune response. AB - Genetic and biochemical dissection of signaling pathways regulating plant pathogen defense has revealed remarkable similarities with the innate immune system of mammals and Drosophila. Numerous plant proteins resembling eukaryotic receptors have been implicated in the perception of pathogen-derived signal molecules. Receptor-mediated changes in levels of free calcium in the cytoplasm and production of reactive oxygen species and nitric oxide constitute early events generally observed in plant-pathogen interactions. Positive and negative regulation of plant pathogen defense responses has been attributed to mitogen activated protein kinase cascades. In addition, salicylic acid, jasmonic acid and ethylene are components of signaling networks that provide the molecular basis for specificity of plant defense responses. This article reviews recent advances in our understanding of early signaling events involved in the establishment of plant disease resistance. PMID- 11495792 TI - PCI complexes: pretty complex interactions in diverse signaling pathways. AB - Three protein complexes (the proteasome regulatory lid, the COP9 signalosome and eukaryotic translation initiation factor 3) contain protein subunits with a well defined protein domain, the PCI domain. At least two (the COP9 signalosome and the lid) appear to share a common evolutionary origin. Recent advances in our understanding of the structure and function of the three complexes point to intriguing and unanticipated connections between the cellular functions performed by these three protein assemblies, especially between translation initiation and proteolytic protein degradation. PMID- 11495796 TI - Cytology of fungal pathogens and plant-host interactions. AB - Imaging plays a unique role in fungal cell biology and phytopathology by allowing for the documentation of molecular structure in individual fixed and living cells. Advances in fluorescence laser techniques, including confocal and multiphoton microscopy, are opening new avenues for cellular exploration. These techniques hold tremendous potential for studies of host-pathogen interactions including the use of genetically encoded markers such as green fluorescent protein, in situ hybridization and fluorescence resonance energy transfer. PMID- 11495797 TI - Fungal gene expression during pathogenesis-related development and host plant colonization. AB - To successfully infect plants, pathogenic fungi must recognize and communicate with their host during different stages of the disease cycle. In past years, techniques such as insertional mutagenesis, sensitive GFP-based reporter systems and microarray techniques have been developed to analyze these processes at the molecular level, and now novel insights into this fascinating aspect of pathogen plant communication are beginning to emerge. This is exemplified by a number of pathogenicity genes functioning in distinct stages of pathogenic development in Magnaporthe grisea. PMID- 11495798 TI - Gene information for fungal plant pathogens from expressed sequences. AB - Gaining information on global gene function in fungal plant pathogens through analysis of expressed sequences is a valuable step towards understanding the overall metabolism and biology underlying the pathogenic habit. Such large-scale gene-finding exercises are poised to provide many useful leads for discovering key events that contribute to the processes of infection and colonisation of host plants. PMID- 11495799 TI - Functional analysis of pathogenicity genes in a genomics world. AB - Genome-wide mutational and expression analyses have been performed in yeast and provide a model for large-scale analysis of gene function in filamentous fungi. The recent completion of the Neurospora crassa genome offers a resource for comparative analysis with plant pathogenic filamentous fungi. These advances have important implications for molecular genetic studies of pathogenicity genes. PMID- 11495800 TI - Epichloe endophytes: fungal symbionts of grasses. AB - Recent developments have increased our understanding of the evolution of mutualistic associations between Epichloe endophytes and their grass hosts. Most of the asexual species appear to be interspecific hybrids. Although endophytes form compatible associations with their natural hosts, transfers to other hosts elicit a range of incompatible reactions. Recently, the genes involved in the synthesis of the secondary metabolites ergot alkaloids and indole-diterpenes, which confer protective benefits on the association, have been cloned. PMID- 11495802 TI - Trypanosome lytic factors: novel mediators of human innate immunity. AB - A novel trypanosome lytic factor (TLF) has been characterized that protects humans from infection by Trypanosoma brucei brucei. The mechanism of trypanolysis is unknown; contrary to one hypothesis, TLF does not kill trypanosomes by generating oxygen radicals. However, these trypanosomes become human-infective when they express a serum-resistance-associated gene. PMID- 11495803 TI - Plasmodium telomeres: a pathogen's perspective. AB - New data on the organization of plasmodial telomeres has recently become available. Telomeres form clusters of four to seven heterologous chromosome ends at the nuclear periphery in asexual and sexual parasite stages. This subnuclear compartment promotes gene conversion between members of subtelomeric virulence factor genes in heterologous chromosomes resulting in diversity of antigenic and adhesive phenotypes. This has important implications for parasite survival. PMID- 11495804 TI - Conservation of a novel vacuolar transporter in Plasmodium species and its central role in chloroquine resistance of P. falciparum. AB - Chloroquine resistance in Plasmodium falciparum has recently been shown to result from mutations in the novel vacuolar transporter, PfCRT. Field studies have demonstrated the importance of these mutations in clinical resistance. Although a pfcrt ortholog has been identified in Plasmodiumvivax, there is no association between in vivo chloroquine resistance and codon mutations in the P. vivax gene. This is consistent with lines of evidence that suggest alternative mechanisms of chloroquine resistance among various malaria parasite species. PMID- 11495805 TI - Encystation in parasitic protozoa. AB - Giardia and Entamoeba parasites encase themselves in a carbohydrate-rich cyst for travel from host to host. Both parasites upregulate their Golgi apparatus during this process, yielding organelles that are now found to be similar to those of higher eukaryotes. In fact, unusual enzymes and structural proteins used for cyst wall synthesis, the complexity of the secretory pathways used to transport materials to the developing cyst walls, as well as unexpected mechanisms of gene regulation and parasite-host and parasite-parasite information exchange, are revealing a high level of sophistication in these organisms that occupy low branch points in the eukaryotic lineage. PMID- 11495806 TI - Protist tubulins: new arrivals, evolutionary relationships and insights to cytoskeletal function. AB - The protists exhibit probably the most extravagant expression of microtubule containing structures found in any organism. These structures--flagella, cilia, axostyles, spindles and a veritable constellation of microtubule bundles and cortical arrays--provide shape, form, motility, anchorage and apparatuses for feeding. The cytoskeletal structures have a precise order (i.e. size, position and number) that must be replicated and segregated with fidelity at each division, some components being inherited conservatively and others semi conservatively. Intriguingly, it is now apparent that much of the high-order organisation, which was recognised and described by light and electron microscopy during the last century, is a reflection of molecular polarities set by assembly of constituent proteins. Tubulins and microtubules lie at the heart of this morphogenetic pattern. PMID- 11495808 TI - Interactions between rotavirus and gastrointestinal cells. AB - Rotaviruses are the leading cause of life-threatening diarrheal disease in infants and in young animals worldwide. The outcome of rotavirus infection of intestinal epithelial cells is more complex and involves induction of more diverse cellular responses than initially appreciated. Similar to bacteria, the pathogenesis of rotavirus-induced disease involves an enterotoxin, activation of the enteric nervous system and malabsorption, suggesting that common mechanisms of pathogenesis may exist between viral and bacterial pathogens. PMID- 11495809 TI - Varicella-zoster virus: molecular virology and virus-host interactions. AB - Cosmid-based mutagenesis and methods to examine varicella-zoster virus (VZV) tropism for differentiated human cells in vivo provide new information about molecular mechanisms of VZV infection. How specific VZV gene products contribute to viral replication has been further defined, and effects of VZV on expression of cellular genes have been demonstrated. PMID- 11495810 TI - The machinery for flavivirus fusion with host cell membranes. AB - A combination of structural, biochemical and functional studies with the flavivirus tick-borne encephalitis virus has revealed the characteristics of a new class of viral fusion protein, class II, that is unrelated to the class I viral fusion proteins for which influenza virus hemagglutinin is the prototype. New structural data have shown that the alphaviruses, another group of icosahedral enveloped viruses, also have class II fusion proteins, suggesting a common origin. PMID- 11495811 TI - Novel virulence and host range genes of African swine fever virus. AB - Current work is beginning to reveal the complex mechanisms by which African swine fever virus interacts with its swine and tick hosts. This work includes the identification of novel viral genes that mediate virulence and host range, and influence important cellular regulatory pathways. PMID- 11495812 TI - Mouse hepatitis virus. AB - Inoculation of mice with most neurotropic strains of the coronavirus mouse hepatitis virus results in an immune response-mediated demyelinating disease that serves as an excellent animal model for the human disease multiple sclerosis. Recent work has shown that either virus-specific CD4(+) or CD8(+) T cells are able to mediate demyelination and also that the antibody response is crucial for clearing infectious virus. Another exciting advance is the development of recombinant coronaviruses, which, for the first time, will allow genetic manipulation of the entire viral genome. PMID- 11495813 TI - Borna disease: virus-induced neurobehavioral disease pathogenesis. AB - Studies of the pathogenesis of neurobehavioral diseases following Borna disease virus infections have been increasing rapidly over the past ten years. Recent major advances have included a report of vertical transmission of the virus in its natural host, the horse, and a report of isolation of a novel variant, No/98, in that same species. In rats infected neonatally with the Borna disease virus that lack blood-borne inflammation in the brain, evidence of an "endogenous" brain inflammatory response is abundant, with elevated expression of cytokine and chemokine mRNA. Infection in these rats is also associated with abnormal levels of neurotransmitters, including serotonin and norepinephrine. Data and debate continue to be forthcoming about the role of Borna disease virus in human infection and psychiatric disease. PMID- 11495814 TI - [Screening for cancer: from guidelines to practice]. PMID- 11495815 TI - [The CCN family of cell growth regulators: a new family of normal and pathologic cell growth and differentiation regulators: lessons from the first international workshop on CCN gene family]. PMID- 11495816 TI - [Leukemogenesis and new therapy development: the example of chronic myelogenous leukemia]. AB - During the past ten years, the improvements of our understanding of cellular signal transduction pathways provide new targets for drug therapies. Chronic myeloid leukemia (CML), a malignant hematopoietic stem cell disorder, is characterised by an acquired genetic abnormality: the Philadelphia chromosome (Ph) and its molecular counterpart, the oncogene BCR-ABL. The latter, which is translated in an active BCR-ABL protein, exhibited a deregulated tyrosine kinase activity inducing malignant transformation. Produced from the 2 phenylaminopyrimidine class, a novel synthetic inhibitor, identified as CGP57148 (STI571), inhibits tyrosine kinase activity of c-ABL, BCR-ABL, PDGF-R and c-kit at micromolar concentrations. It suppresses the proliferation of the majority of BCR-ABL positive cell lines. The phases I-II clinical trials in CML have demonstrated promising results, especially in the chronic phase of the disease. STI571 is an original therapeutic approach which may be used as a model for the development of other drugs in cancer. PMID- 11495817 TI - [Anti-tyrosine kinase: the beginning of molecular therapies of cancer and initial results]. PMID- 11495818 TI - [STI571 and gastro intestinal stromal tumors]. PMID- 11495819 TI - [Proteomic analysis: why and how ?]. AB - The proteome, first formalized in 1995, designs all the proteins expressed by the genome of a cell, tissu, or organ at a defined time. Proteomic analysis leads to a description of the regulation of gene expression by the study of proteins and of their post-translational modifications. Proteomic analysis is based on three technologies: 1) Two-dimensional electrophoresis allowing the separation of thousands of proteins from a single mixture; 2) mass spectrometry allowing the characterization of picoquantities of polypeptides and providing data on post translational modifications; 3) Bioinformatic which is required for the quantification of protein level and for the constitution of databases of protein expression profiles. Complementing the methods of the genomics, the use of proteomic analysis is widely spreading in the fields of fundamental biology, biomedicine and pharmacology for the identification of new biological markers and therapeutic targets. PMID- 11495820 TI - [What do we know about ATM protein expression in breast tissue?]. AB - The great majority of breast cancer cases are not associated with a mutated gene of high penetrance such as BRCA1, BRCA2 and TP53. Genes of low penetrance, frequently mutated in the general population, might play an important role in breast cancer development. The ATM gene, which encodes the ATM protein, mutated in the disorder ataxia telangiectasia (AT) could be such a susceptibility gene. Indeed, 1% of the general population is estimated to be AT heterozygote and females have an increased risk of developing breast cancer. The ATM protein is involved in the signalling pathway of DNA double-strand breaks. Studies on its expression in normal breast tissues have shown that ATM is expressed in the epithelial cells of breast ducts, but not in the myoepithelial cells. In sclerosing adenosis, a benign lesion of the breast, the ATM protein is expressed in both cell types whereas its expression is absent or reduced in tumour epithelial cells in about 30-50% of invasive carcinomas. Moreover, the study of the p53 status in some of these tumours has revealed that the ATM/p53 signalling pathway is frequently altered either by a very low ATM expression or by the presence of a mutated p53. It remains to be determined whether alterations in the expression of other proteins also involved in this DNA damage signalling cascade are specifically associated with breast cancer development and/or a radiosensitive phenotype seen in some breast cancer patients after radiotherapy. PMID- 11495821 TI - [Medical management of women with inherited predisposition to breast cancer: indications and procedures for mammographic screening]. AB - Women identified or suspected as carriers of mutations in BRCA1 or BRCA2 susceptibility genes have a high risk to develop an early breast cancer and thus, require appropriate management. Some consensus guidelines were provided for women at hereditary risk and two possible strategies of prevention are suggested: breast cancer screening and prophylactic surgery. We present the French recommendations for breast cancer surveillance and discuss the justification, indications and modalities of mammographic screening. Screening by annual mammography is recommended from age 30 years in experienced centers, in association with semi-annual clinical breast examination from age 20 years. These recommendations apply to women who were identified as carriers of a cancer predisposing mutation of BRCA1 or BRCA2 genes. In families for whom any mutation of the two genes could be identified, the same modalities apply also to women with a higher probability than 25% of being a carrier. We present here an illustration of the calculation of such probabilities from two example-pedigrees. PMID- 11495822 TI - [Recommendations on cancer screening in the European Union]. PMID- 11495823 TI - [Local control in the management of cancer: a few thoughts]. AB - Cancer is still considered from a physiopathological point of view as a disease of the cell. This concept is underlying the idea of cure. Treatment with curative intent should aim at eradicating all the tumoral cells. Local control is mandatory and essential in cancers localized in organ with vital function. In breast cancer a complex and controversial relationship exists between local relapse and increased risk of fatal distant metastasis. In case of organ preserving treatment, a complete local control is necessary from the start. If a too high risk of local relapse is foreseable, conservative treatment should not be recommended. PMID- 11495824 TI - [Double-blinded controlled study comparing clodronate versus placebo in patients with breast cancer bone metastases]. AB - One hundred forty-four patients with breast cancer and osteolytic bone metastases were randomized to receive either oral clodronate 1,600 mg/d (73 patients) or placebo (71 patients), in addition to either chemotherapy or hormonal therapy, for up to 12 months. Patients were withdrawn from the study when the 12 months of treatment had been achieve or a new bone event occurred, which was defined as: hypercalcemia (> 3 mmol/l), increase in, or onset of new bone pain due to metastases, requirement of radiotherapy for bone pain relief, pathological fractures (including vertebral collapse, spinal cord compression) or death due to bone metastases. Patients are well balanced according to age, performance status, bone condition, except for fractures, more frequent in the clodronate group (25% vs 12%). Of the 137 evaluable patients, 69 received oral clodronate and 68 placebo. Clodronate significantly delayed the median time to onset of new bone events compared to placebo, respectively 244 days and 180 days (p = 0.05). Hypercalcemia did not occur in the clodronate group but was observed in four placebo-treated patients. Clodronate-treated patients had a significant reduction in pain intensity compared to placebo (p = 0.01; measured using a visual pain scale) and significantly fewer patients receiving clodronate required analgesics (p = 0.02). The evaluation of global efficacy by physicians and patients indicated that clodronate was more efficacious than placebo (respectively p = 0.02 and p = 0.01). No significant difference in incidence of adverse effects was observed between the two groups. Clodronate therapy significantly delayed the occurrence of new bone events in these patients with bone metastases from breast cancer and adds to treatment of malignant osteolysis. PMID- 11495825 TI - [Protontherapy is not an obsolete technic]. PMID- 11495827 TI - What causes specificity of practice in a manual aiming movement: vision dominance or transformation errors? AB - The withdrawal of vision of the arm during a manual aiming task has been found to result in a large increase in aiming error, regardless of the amount of practice in normal vision before its withdrawal. In the present study, the authors investigated whether the increase in error reflects the domination of visual afferent information over the movement representation developed during practice to the detriment of other sources of afferent information or whether it reflects only transformation errors of the location of the target from an allocentric to an egocentric frame of reference. Participants (N = 40) performed aiming movements with their dominant or nondominant arm in a full-vision or target- only condition. The results of the present experiment supported both of those hypotheses. The data indicated that practice does not eliminate the need for visual information for optimizing movement accuracy and that learning is specific to the source or sources of afferent information more likely to ensure optimal accuracy during practice. In addition, the results indicated that movement planning in an allocentric frame of reference might require simultaneous vision of the arm and the target. Finally, practice in a target-only condition, with knowledge of results, was found to improve recoding of the target in an egocentric frame of reference. PMID- 11495828 TI - Influence of strategy on muscle activity during impact movements. AB - Participants (N = 10) made flexions or extensions about the elbow. Movements either were pointing (i.e., self-terminated) or terminated by impact on a barrier. The author examined how the trajectory and the electromyographic (EMG) patterns varied according to the distance moved, the instruction provided concerning speed, or the type of termination. Variations in kinematics induced by changes in the target distance or the instruction regarding speed were the same for impact and pointing movements. In comparison with a pointing movement of similar distance and speed instruction, an impact movement (a) accelerated longer and reached a higher velocity, (b) had a longer agonist EMG burst, and (c) had a low level of contraction that started slightly after the agonist burst and continued throughout the movement but had little or no antagonist burst. Because the different types of movements required different forces from the muscles, there were systematic, task-specific differences in EMG patterns that reflected task-specific differences in central control. The results of this experiment demonstrate that impact movements share some of the rules used in the control of other tasks, such as pointing and reversing movements. The sharing is not imposed by mechanical or physiological constraints but, rather, represents the imposition of internal constraints. PMID- 11495829 TI - Development of multiple movement representations with practice: specificity versus flexibility. AB - The question addressed in the present experiment was whether an individual who practices a task under different conditions of afferent information develops different movement representations, each of which is based on the most accurate source of afferent information for movement control. In Experiment 1, participants (N = 23) performed a manual aiming movement in a target-only condition for 520 trials before performing in a normal vision condition for an equivalent amount of practice. Control groups performed all practice trials in either a normal vision or a target-only condition. The results revealed that the movement representation developed in the initial (target-only) practice phase remained accessible for movement planning and control. The results of Experiment 2 indicated, however, that participants did not maintain such a representation when their initial practice in the target-only condition was reduced (40 or 160 trials) before they had extensive practice in normal vision. Those results indicate that extensive practice in a target-only and then in a normal vision condition enables an individual to plan and control his or her movement on the basis of the most efficient source of available afferent information. Because visual afferent information provides optimal information for ensuring movement accuracy, however, if initial practice in the target-only condition is only modest or moderate it is likely that that information source will progressively dominate all other sources of afferent information for movement planning and control. PMID- 11495830 TI - Planning of reach-and-grasp movements: effects of validity and type of object information. AB - Individuals are assumed to plan reach-and-grasp movements by using two separate processes. In 1 of the processes, extrinsic (direction, distance) object information is used in planning the movement of the arm that transports the hand to the target location (transport planning); whereas in the other, intrinsic (shape) object information is used in planning the preshaping of the hand and the grasping of the target object (manipulation planning). In 2 experiments, the authors used primes to provide information to participants (N = 5, Experiment 1; N = 6, Experiment 2) about extrinsic and intrinsic object properties. The validity of the prime information was systematically varied. The primes were succeeded by a cue, which always correctly identified the location and shape of the target object. Reaction times were recorded. Four models of transport and manipulation planning were tested. The only model that was consistent with the data was 1 in which arm transport and object manipulation planning were postulated to be independent processes that operate partially in parallel. The authors suggest that the processes involved in motor planning before execution are primarily concerned with the geometric aspects of the upcoming movement but not with the temporal details of its execution. PMID- 11495831 TI - Speed modulation in swimming frogs. AB - Swimming movements of 7 European green frogs (Rana esculenta) were studied, starting from the detailed analysis of the speed and timing of the propulsive, glide, and recovery phases of their intermittent swimming behavior. First, the authors identified the spatiotemporal factors used by the frogs to modulate their swimming behavior. None of the gait variables correlated strongly with average swimming speed, and no significant correlations were found between variables belonging to different phases. There did not seem to be an obvious control strategy. Instantaneous speeds at the transition of the different phases all increased significantly with average speed, however. The strong correlation between maximal speed at the end of propulsion and the speed averaged over a cycle might reflect the dominance of the propulsive phase in the determination of the overall swimming speed. The modulation of swimming speed thus seemed largely comparable with the regulation of jumping distance. That finding was confirmed in a mathematical model, in which the positive correlations between both glide and recovery speeds, on the one hand, and average speed, on the other, were shown to be only mathematical consequences of the strong impact of the propulsive phase on overall swimming performance. That finding suggests that the correlations did not result from an active control strategy. PMID- 11495832 TI - Planning and executing simple movements: contributions of relative-time and overall-duration specification. AB - In 3 experiments, the authors used a precuing protocol to examine the nature and cost of programming and the subsequent reprogramming of a movement's relative time and overall duration. Initial programming followed a fixed-order specification; knowledge of the necessary relative time was required before information regarding overall duration could be used in a manner that expedited response planning. In the case of reprogramming, however, when a modification had to be made in either the relative time or overall duration of the anticipated and already-prepared response, performers chose to completely reprogram the entire response. Complete reprogramming occurred even when the performer had correctly prepared the higher order relative-time component and only had to modify the overall duration of the movement. The data indicate that organizing movement timing before movement initiation is accomplished in a fundamentally different manner depending on whether the movement is being initially compiled or modified. PMID- 11495833 TI - Simulating the impact during human jumping by means of a 4-degrees-of-freedom model with time-dependent properties. AB - The authors simulated the vertical movements of a jumper and the force time courses by means of a 4-degrees-of-freedom model consisting of 4 masses, springs, and dampers. Of the motions simulated, only that of the mass imitating the trunk corresponded to the measured data. The best fit to the measured force curves were obtained in the simulation in which time-dependent model parameters were used. From the results, the authors concluded that at the beginning of the landing, a jumper behaves like a 2-mass model in which the leg segments (thighs, shanks, and feet) effectively combine into 1 mass. After approximately 60 ms, the connections between the leg segments become more compliant and the jumper behaves like a 4 mass model with a soft coupling between the leg segments. The process is equivalent to an increase of the degrees of freedom of the movements. At the end of the ground contact phase during hopping, the jumper has to contract the muscles in order to reach the envisaged jump height. In the model, that contraction could not be satisfactorily simulated. PMID- 11495834 TI - Specificity of task constraints and effects of visual demonstrations and verbal instructions in directing learners' search during skill acquisition. AB - In the present study, the efficacy of visual demonstrations and verbal instructions as instructional constraints on the acquisition of movement coordination was investigated. Fifteen participants performed an aiming task on 100 acquisition and 20 retention trials, under 1 of 3 conditions: a modeling group (MG), a verbally directed group (VDG), and a control group (CG). The MG observed a model intermittently throughout acquisition, whereas the VDG was verbally instructed to use the model's movement pattern. Participants in the CG received neither form of instruction. Kinematic analysis revealed that compared with verbal instructions or no instructions, visual demonstrations significantly improved participants' approximation of the model's coordination pattern. No differences were found in movement outcomes. Coordination data supported the visual perception perspective on observational learning, whereas outcome data suggested that the modeling effect is mainly a function of task constraints, that is, the novelty of a movement pattern. PMID- 11495835 TI - Visual information and the control of reaching in children: a comparison between children with and without developmental coordination disorder. AB - Three experiments were performed on reach and grasp in 9- to 10-year-old children (8 controls and 8 with developmental coordination disorder [DCD]). In normal reaching, children in the DCD group were less responsive to the accuracy demands of the task in controlling the transport component of prehension and spent less time in the deceleration phase of hand transport. When vision was removed as movement began, children in the control group spent more time decelerating and reached peak aperture earlier. Children in the DCD group did not do that, although, like the control group, they did increase grip aperture in the dark. When depth cues were reduced and only the target or only the target and hand were visible, children in the control group used target information to maintain the same grip aperture in all conditions, but DCD children behaved as if the target was not visible. Throughout the studies, the control group of 9- to 10-year-olds did not produce adult-like adaptations to reduced vision, suggesting that they had not yet attained adult-like integration of sensory input. Compared with control children, children with DCD did not exhibit increased dependence on vision but showed less recognition of accuracy demands, less adaptation to the removal of vision, and less use of minimal visual information when it was available. PMID- 11495850 TI - Commentary: considerations for use of racial/ethnic classification in etiologic research. AB - Numerous authors have critiqued the use of race as an etiologic quantity in medical research. Despite this criticism, the use of variables encoding racial/ethnic categorization has increased in epidemiology, and most researchers agree that important variation in disease risk is captured by this classification system. Previous discussions have generally neglected to articulate guidelines for appropriate use of racial/ethnic information in etiologic research. The authors summarize the logical, conceptual, and practical problems associated with the "ethnic paradigm" as currently applied in biomedical sciences and offer a set of methodological recommendations toward more valid use of racial/ethnic classification in etiologic studies. These suggested guidelines address issues of variable definition, study design, and covariate control, providing a consistent foundation for etiologic research programs that neither ignore racial/ethnic disease disparities nor obfuscate the nature of these disparities through inappropriate analytical approaches. This methodological analysis of racial/ethnic classification as an epidemiologic quantity provides a formal basis for a focus on racism (i.e., social relations) rather than race (i.e., innate biologic predisposition) in the interpretation of racial/ethnic "effects." PMID- 11495851 TI - Invited commentary: "race," racism, and the practice of epidemiology. PMID- 11495853 TI - Trends in preterm birth and neonatal mortality among blacks and whites in the United States from 1989 to 1997. AB - Preterm birth, a major determinant of infant mortality, has been increasing in recent years. The authors examined trends in preterm birth and its determinants by using the US birth and infant death files for 1989-1997. The impact of trends in preterm birth rates on neonatal and infant mortality was also evaluated. Among Whites, preterm births (<37 completed weeks of gestation) increased from 8.8% of livebirths in 1989 to 10.2% in 1997, a relative increase of 15.6%. On the other hand, preterm births among Blacks decreased by 7.6% (from 19.0% to 17.5%) during the same period. An increase in obstetric interventions contributed to increases in preterm births for both races but was outweighed by other unidentified favorable influences for Blacks. Neonatal mortality among preterm Whites dropped 34% during the 8 years of the study, while the decrease was only 24% among Blacks. This large disparity countered the changes in preterm birth rates so that the percentage decline in neonatal mortality was similar in the two racial groups (18-20%). In conclusion, the anticipated mortality benefit from a lower preterm birth rate for Blacks has been blunted by suboptimal improvement in mortality among the remaining preterm infants. The widening race gap in mortality among preterm infants merits attention. PMID- 11495854 TI - Infertility among women exposed prenatally to diethylstilbestrol. AB - Although it is well established that women exposed to diethylstilbestrol in utero have an increased risk of spontaneous abortion, ectopic pregnancy, and preterm delivery, it is not known whether they also have an increased risk of infertility. The authors assessed this question in data from a collaborative follow-up study of the offspring of women who took diethylstilbestrol during pregnancy. In 1994, 1,753 diethylstilbestrol-exposed and 1,050 unexposed women from an ongoing cohort study (National Cooperative Diethylstilbestrol Adenosis Study and Dieckmann cohorts) provided data on difficulties in conceiving and reasons for the difficulty. Age-adjusted relative risks were computed for the association of diethylstilbestrol exposure with specific types of infertility. A greater proportion of exposed than unexposed women were nulligravid (relative risk (RR) = 1.3, 95% confidence interval (CI): 1.1, 1.5), and a greater proportion had tried to become pregnant for at least 12 months without success (RR = 1.8, 95% CI: 1.6, 2.1). Diethylstilbestrol exposure was significantly associated with infertility due to uterine and tubal problems, with relative risks of 7.7 (95% CI: 2.3, 25) and 2.4 (95% CI: 1.2, 4.6), respectively. The present findings indicate that diethylstilbestrol-exposed women have a higher risk of infertility than do unexposed women and that the increased risk of infertility is primarily due to uterine or tubal problems. PMID- 11495855 TI - Exposure to tobacco smoke in utero and the risk of stillbirth and death in the first year of life. AB - The authors examined the association between exposure to tobacco smoke in utero and the risk of stillbirth and infant death in a cohort of 25,102 singleton children of pregnant women scheduled to deliver at Aarhus University Hospital, Aarhus, Denmark, from September 1989 to August 1996. Exposure to tobacco smoke in utero was associated with an increased risk of stillbirth (odds ratio = 2.0, 95% confidence interval: 1.4, 2.9), and infant mortality was almost doubled in children born to women who had smoked during pregnancy compared with children of nonsmokers (odds ratio = 1.8, 95% confidence interval: 1.3, 2.6). Among children of women who stopped smoking during the first trimester, stillbirth and infant mortality was comparable with that in children of women who had been nonsmokers from the beginning of pregnancy. Conclusions were not changed after adjustment in a logistic regression model for the sex of the child; parity; or maternal age, height, weight, marital status, years of education, occupational status, and alcohol and caffeine intake during pregnancy. Approximately 25% of all stillbirths and 20% of all infant deaths in a population with 30% pregnant smokers could be avoided if all pregnant smokers stopped smoking by the sixteenth week of gestation. PMID- 11495856 TI - Small-area variations and sociodemographic correlates for the incidence of Crohn's disease and ulcerative colitis. AB - The objectives of this study were to describe variations in the incidence of inflammatory bowel disease (IBD) within the Canadian province of Manitoba and to analyze sociodemographic factors associated with these variations. The authors used the Manitoba Health insurance databases to measure incidence rates of Crohn's disease and ulcerative colitis for each of 52 postal areas in Manitoba, in 1987-1996. The sociodemographic characteristics of the postal areas were based on data from the 1996 Canadian census. The overall incidence rates of Crohn's disease and ulcerative colitis were identical-15.6 per 100,000. Both diseases showed substantial geographic variation, with incidence rates differing significantly from the provincial average in 15 postal areas for Crohn's disease and in 13 postal areas for ulcerative colitis. There was a significant geographic correlation in the incidence of Crohn's disease and ulcerative colitis (r = 0.49, p < 0.001). The incidence of IBD was higher in urban areas (incidence rate ratio (IRR) = 1.21, 95% confidence interval (CI): 1.00, 1.45). Aboriginal Canadians had significantly lower rates of both Crohn's disease (IRR = 0.11, 95% CI: 0.05, 0.22) and ulcerative colitis (IRR = 0.57, 95% CI: 0.42, 0.79). A higher incidence of IBD was ecologically associated with a higher average family income, a lower proportion of immigrant and Aboriginal Canadian populations, and a smaller average family size. PMID- 11495857 TI - Case-control study of lifetime physical activity and breast cancer risk. AB - A population-based case-control study of 1,233 incident breast cancer cases and 1,237 controls was conducted in Alberta, Canada, in 1995-1997 to examine the effect of lifetime physical activity patterns on breast cancer risk. No associations between physical activity and breast cancer were found for premenopausal women. For postmenopausal women in the highest quartile (> or =161 metabolic equivalent (MET)-hours/week per year) versus the lowest quartile (<104.8 MET-hours/week per year) of lifetime total physical activity, the adjusted odds ratio was 0.70 (95% confidence interval (CI): 0.52, 0.94). When the risks associated with each type of activity were examined for postmenopausal women, household and occupational activity conferred the largest risk reductions (odds ratio (OR) = 0.57, 95% CI: 0.41, 0.79 and OR = 0.59, 95% CI: 0.44, 0.81, respectively, for highest vs. lowest quartiles of activity), while recreational activity was not associated with any risk reductions. For postmenopausal women, the authors found stronger risk reductions for those who were also nonsmokers (OR = 0.64, 95% CI: 0.46, 0.88), non-alcohol-drinkers (OR = 0.39, 95% CI: 0.11, 0.77), or nulliparous (OR = 0.22, 95% CI: 0.07, 0.70) when they compared the highest with the lowest quartile of lifetime total physical activity. This study provides evidence that lifetime total activity reduces risk of postmenopausal breast cancer. PMID- 11495859 TI - Toxoplasma gondii infection in the United States: seroprevalence and risk factors. AB - Infection with Toxoplasma gondii can cause severe illness when the organism is contracted congenitally or when it is reactivated in immune-suppressed persons. To determine the prevalence of T. gondii infection in a representative sample of the US population, the authors tested sera from participants in the Third National Health and Nutrition Examination Survey (1988-1994) for immunoglobulin G antibodies to T. gondii. Of 27,145 persons aged > or =12 years, 17,658 (65%) had sera tested. The overall age-adjusted seroprevalence was 22.5% (95% confidence interval (CI): 21.1, 23.9); among women aged 15-44 years, seroprevalence was 15.0% (95% CI: 13.2, 17.0). Age-adjusted seroprevalence was higher in the Northeast (29.2%) than in the South (22.8%), Midwest (20.5%), or West (17.5%) (p < 0.05). In multivariate analysis, risk for T. gondii infection increased with age and was higher among persons who were foreign-born, persons with a lower educational level, those who lived in crowded conditions, and those who worked in soil-related occupations, although in subset analyses risk categories varied by race/ethnicity. Nearly one quarter of adults and adolescents in the United States have been infected with T. gondii. Most women of childbearing age in the United States are susceptible to acute infection and should be educated about ways to minimize exposure to T. gondii. PMID- 11495858 TI - Correlates of markers of oxidative status in the general population. AB - Oxidative stress has been implicated in the etiology of many chronic diseases, including cardiovascular disease. However, limited information exists on the factors that may influence oxidative status in the general population. In a random sample of the population of two counties in western New York, levels of several markers of oxidative status (i.e., thiobarbituric acid-reactive substances, erythrocyte glutathione, and glutathione peroxidase) were determined. A total of 894 men and 903 women aged 35-79 years were included in the study (1996-1999). In addition, a number of sociodemographic and lifestyle characteristics and cardiovascular disease risk factors were measured. Age, markers of glucose metabolism (e.g., plasma glucose level) and insulin resistance (e.g., serum triglycerides, high density lipoprotein cholesterol, body mass index), and postmenopausal status in women were associated with increased oxidative stress and reduced antioxidant potentials. Oxidative status and antioxidant potentials appear to be significantly associated with a number of major cardiovascular disease risk factors; most of them are linked to abnormalities in glucose and insulin metabolism. PMID- 11495860 TI - Analysis of dynamic cohort data. AB - Left-truncated and interval-censored data, termed dynamic cohort data, arise in longitudinal studies with rolling admissions and only occasional follow-up. The authors compared four approaches for analyzing such data: a constant hazard model; maximum likelihood estimation with flexible parametric models; the midpoint method, in which the midpoint of the last negative and first positive test result is used in a Cox proportional hazards model that accounts for left truncation; and a semiparametric method that uses imputed failure times in the Cox model. By using a simulation study, they assessed the performance of these approaches under conditions that can arise in observational studies: changes in disease incidence and changes in the underlying population. The simulation results indicated that the constant hazard model and midpoint method were inadequate and that the flexible parametric model was useful when enough parameters were used in modeling the baseline hazard. The semiparametric method ensured correct parameter (odds ratio) estimation when the baseline hazard was misspecified, but the trade-off increased computational complexity. In this paper, a study of the incidence of human immunodeficiency virus in patients repeatedly tested for the virus at a sexually transmitted disease clinic in New Orleans, Louisiana, illustrates the methods used. PMID- 11495861 TI - Socioeconomic, demographic, occupational, and health factors associated with participation in a long-term epidemiologic survey: a prospective study of the French GAZEL cohort and its target population. AB - The purpose of this paper is to examine personal and health factors, both at the beginning of the study and thereafter, associated with participation in the GAZEL cohort, set up in 1989 in a large French company. The authors used logistic regression to analyze the associations between participation and data available for both participants (n = 20,093) and nonparticipants (n = 24,829). Higher participation was associated with male sex, marriage, children, managerial status, and residence in particular regions. Among men, lower participation was associated with sick leave in the year before recruitment and afterwards. During follow-up, participation was negatively associated with several groups of diseases, especially those associated with alcohol consumption. The risk of upper respiratory and digestive tract and lung cancer during follow-up was higher among nonparticipants. The same phenomenon occurred among women, but less markedly, for cancers of the breast and genital organs. During follow-up, mortality among men was higher among nonparticipants, especially for alcohol-related diseases. The association among women was less strong. Among men, but not among women, diseases caused by alcohol, smoking, or dangerous behavior were the primary reason for the health differences observed between participants and nonparticipants. Overall, the most important determinants of participation were cultural factors and lifestyle behaviors. PMID- 11495862 TI - Re: "Mortality reduction with air bag and seat belt use in head-on passenger car collisions". PMID- 11495865 TI - [Fat embolism and postoperative coagulopathy]. PMID- 11495866 TI - Fast-tracking in ambulatory anesthesia. PMID- 11495867 TI - Fat embolization and fatal cardiac arrest during hip arthroplasty with methylmethacrylate. AB - PURPOSE: This case report describes a cardiac arrest during a cemented hip arthroplasty procedure. Hemodynamic instability during methylmethacrylate use in arthroplasty surgery can be explained by fat embolization rather than the inherent toxicity of the monomer. CLINICAL FEATURES: A 78-yr-old woman required a cemented hemiarthroplasty for a pathologic left subcapital fracture. The patient's past medical history included stable angina, diet-controlled type II diabetes and metastatic breast cancer. During the cementing of the canal and insertion of the femoral prosthesis, desaturation, hypotension and cardiac arrest occurred. The patient underwent a successful intraoperative resuscitation and was transferred to the intensive care unit where she subsequently developed disseminated intravascular coagulopathy. The patient died 24 hr later and autopsy confirmed the cause of death as fat embolization. CONCLUSION: The deleterious cardiovascular effects of methylmethacrylate have been discussed in the literature. However, clinical evidence supports fat embolization during arthroplasty surgery as a greater determinant of hemodynamic compromise. Surgical precautions are paramount in minimizing the sequelae of Bone Implantation Syndrome and anesthetic treatment consists of supportive care. PMID- 11495868 TI - A pilot study of recovery room bypass ("fast-track protocol") in a community hospital. AB - PURPOSE: To evaluate the effectiveness of short-acting anesthetic drugs and techniques to achieve recovery room bypass criteria after minor surgery in a community hospital environment. METHODS: After agreement by a multidisciplinary committee, a pilot project was undertaken to assess the usefulness of ultra- short acting anesthetic drugs and pre-emptive analgesia to facilitate rapid recovery from general anesthesia. A cohort of 100 ASA I-II patients aged 18-65 yr undergoing simple knee arthroscopy or minor peripheral orthopedic procedures was compared to a similar cohort treated in the three months prior to the study period. Outcomes of interest included patient morbidity, success in achieving post-anesthesia care unit (PACU) bypass criteria, impact upon nursing resources, duration of operating room (OR) and hospital stay, and pharmaceutical costs before and after implementation. RESULTS: No patient morbidity was demonstrated prior to discharge home, and successful PACU bypass occurred in 83% of cases. Achievement of PACU discharge criteria while in the OR did not prolong the OR time, and discharge from hospital occurred earlier in the patients who did not require PACU care (P=0.0006 all "fast-track cases" vs all "controls"). Nursing complaints were more numerous when the day surgery personnel did not normally participate in PACU care. The cost of anesthetic care was significantly more using ultra-short acting drugs (CDN $14.17 vs CDN $20.57), but closer adherence to protocol could reduce this differential (CDN $18.84). CONCLUSION: Not all patients who receive a general anesthetic require admission to a phase I recovery facility. However, the justification for use of more expensive pharmaceuticals to achieve PACU bypass requires extensive changes in operating systems and voluntary professional behaviours. PMID- 11495869 TI - A survey of professional satisfaction among Canadian anesthesiologists. AB - PURPOSE: To assess overall job satisfaction among Canadian anesthesiologists and examine contributing factors. METHODS: A confidential postal survey of all active members of the Canadian Anesthesiologists Society was conducted in 1998. Demographics, anesthesia practice, overall job satisfaction, anesthetic assistance, and perceived surgeons' and public attitudes towards anesthesiologists were collected. RESULTS: Of 1659 surveys sent, 946 were returned (57% response rate). Seventy-five percent of the respondents were male and 25% female. Staff comprized 91%, residents 9%. The average working week was 59 +/- 11.9 hr. Seventy-five percent of respondents reported overall job satisfaction. Job satisfaction was associated with intellectual stimulation, good quality of care and interaction with patients. Dissatisfaction stemmed from treatment from the provincial government, hospital politics and long hours. Job satisfaction was associated with satisfaction with the level of operating room (OR) assistance, perceived high surgical regard and public image. Residents were more satisfied than staff anesthesiologists. Overall satisfaction was not affected by age, gender, region of practice, type of hospital or clinical work. CONCLUSIONS: Job satisfaction among anesthesiologists is significantly associated with intellectual stimulation, quality of care, interaction with the patients, treatment from the provincial government, hospital politics, working hours, OR assistance and perceived attitude of surgeons. Improving these contributing factors may lead to higher job satisfaction. PMID- 11495870 TI - Sevoflurane with remifentanil allows rapid tracheal intubation without neuromuscular blocking agents. AB - PURPOSE: After inhalational induction with sevoflurane, we compared the effects of adding remifentanil 1 microg x kg(-1) or remifentanil 2 microg x kg(-1) on conditions for tracheal intubation without neuromuscular blocking agents. METHODS: Before anesthetic induction, all patients were given 0.2 mg of glycopyrrolate iv to counteract the bradycardic effects of remifentanil. Two minutes after inhalational induction with 8% sevoflurane and 50% nitrous oxide, 56 female patients with normal airways scheduled for gynecologic surgery were randomized to receive remifentanil 1 or 2 microg x kg(-1) in a double-blind fashion. One minute later, laryngoscopy was initiated for tracheal intubation. Conditions for tracheal intubation and hemodynamic response to tracheal intubation were assessed. RESULTS: Tracheal intubation was successful in all patients. The incidence of post-intubation coughing was lower in the remifentanil 2 microg x kg(-1) group compared to remifentanil 1 microg x kg(-1) group (11% vs 39%, P <0.02). Optimal intubation conditions were also higher in the remifentanil 2 microg x kg(-1) group at 89% vs 54% (P <0.01). However, the higher dose of remifentanil also resulted in a greater decrease in mean arterial pressure (P <0.05). CONCLUSIONS: The addition of remifentanil after sevoflurane induction allows for rapid tracheal intubation without neuromuscular blocking agents. The higher dose of remifentanil results in improved conditions for tracheal intubation but also caused a greater decrease in mean arterial pressure. Tracheal intubation using sevoflurane and remifentanil may be an alternative to traditional tracheal intubation with neuromuscular blocking agents. PMID- 11495871 TI - Xenon inhalation increases norepinephrine release from the anterior and posterior hypothalamus in rats. AB - PURPOSE: To investigate the effect of xenon (Xe) and nitrous oxide (N(2)O) on norepinephrinergic neuronal activity in the rat medial preoptic area (mPOA) and posterior hypothalamus (PH) using microdialysis. METHODS: Sixty male Wistar rats were equally allocated to two groups: mPOA and PH. A microdialysis probe was implanted into the mPOA or the PH. In both groups, each animal was exposed to one of the following inhalations: 25% oxygen (control, n=6), 30% Xe (n=6), 60% Xe (n=6), 30% N(2)O (n=6) or 60% N(2)O (n=6). Norepinephrine concentration in the perfused artificial cerebrospinal fluid was measured by high pressure liquid chromatography at ten-minute intervals. After plotting the time-norepinephrine concentration curve, the area under the curve (AUC) in each group was calculated. RESULTS: In the mPOA, 30 and 60% Xe, but only 60% N(2)O significantly increased norepinephrine release. The AUC in the 30% Xe, 60% Xe or 60% N(2)O group was 160 +/- 9 (P <0.05), 288 +/- 42 (P <0.01) or 237 +/- 46 pg x min/sample (P <0.01), respectively, compared to that in the control group: 77 +/- 14 pg x min/sample. In the PH, only 60% Xe significantly increased norepinephrine release compared to control (AUC: 191 +/- 38 vs. 71 +/- 1 pg x min/sample, P <0.01). CONCLUSION: The present data suggest that Xe stimulates norepinephrinergic neurons more potently than N(2)O; 1.2 times more in the mPOA and 2.5 times more in the PH. This stimulant effect may contribute to the hypnotic and sympathotonic effects of Xe in rats. PMID- 11495872 TI - Blocks at the wrist provide effective anesthesia for carpal tunnel release. AB - PURPOSE: Distal blocks are not recommended even for a short procedure when a tourniquet is used. This study was designed to evaluate the tolerance, effectiveness, patient acceptance and safety of distal blocks at the wrist. METHODS: Consecutive patients (n=273, mean age 53 +/-15 yr) undergoing endoscopic carpal tunnel release with a pneumatic tourniquet were included in this study. The median nerve was blocked 6 cm above the wrist crease by injecting 10 mL of 2% lidocaine and 0.5% bupivacaine (v/v). The ulnar nerve was blocked by injecting 8 mL of the same anesthetic mixture below the flexor carpi ulnaris tendon 6 cm above the wrist crease. Finally, 2 mL of local anesthetic were infiltrated sc and laterally below the crease to block the musculocutaneous nerve. The intensity of the block was evaluated after five, ten and 20 min. In addition, pain associated with block performance and tolerance of the tourniquet were evaluated. Finally, neurological complications associated with this technique were investigated. Data are presented as means +/- SD. RESULTS: At ten minutes after the block was performed, 9% and 32% of patients required an additional injection to complete the block in the median and ulnar territories, respectively. In more than 75% of patients, performance of the block was associated with either no or mild pain. The tourniquet was inflated for 12.6 +/- 5.4 min and was well tolerated in 99% of patients. Finally, neither transient nor permanent neurological deficit were recorded postoperatively. CONCLUSION: Blocks at the wrist are effective, well accepted by the patient and safe when a pneumatic tourniquet is used for a short procedure. PMID- 11495873 TI - Preoperative diclofenac is a useful adjunct to spinal anesthesia for day-case varicose vein repair. AB - PURPOSE: To examine if preoperative diclofenac 50 mg or diazepam 10 mg po are useful adjuncts to spinal anesthesia for day-case varicose vein repair. METHODS: Two hundred ASA physical status I-II outpatients, age 18-60 yr, were randomized to receive either diclofenac 50 mg po or diazepam 10 mg po one hour before operation in a double-blind fashion (100 patients in both groups). If the patient was distressed or feared the spinal puncture and requested sedation, a bolus dose of alfentanil 0.5 mg was given i.v. as a rescue medication. On request, patients received diclofenac 50 mg po and, when needed, oxycodone 0.1 mg x kg(-1) im for postoperative pain relief. They were discharged with a supply of diclofenac 50 mg tablets and were asked to record postoperative pain using a visual analogue scale (VAS) and quantity of tablets taken. RESULTS: The VAS scores (+/- SD) eight hours after surgery, the next morning, and in the morning and at the end of the first and second postoperative days were 23 +/- 21, 12 +/- 17, 11 +/- 15, 8 +/- 15 and 8 +/- 15 in the diclofenac group, and 24 +/- 23, 12 +/- 20, 10 +/- 17, 8 +/- 16 and 7 +/- 14 in the diazepam group, respectively (NS). In the diclofenac and diazepam groups, 31% and 67% of the patients required postoperative diclofenac during the first eight postoperative hours (P <0.05). Diazepam premedication did not alter the number of patients who required alfentanil before spinal puncture. CONCLUSION: Diclofenac premedication reduced the analgesic requirements during the first eight hours after varicose vein repair. PMID- 11495874 TI - A rat model of postthoracotomy pain: behavioural and spinal cord NK-1 receptor assessment. AB - PURPOSE: To develop a new rat model of postthoracotomy pain for investigating its mechanisms and clarifying neurochemical changes. METHODS: Male Wistar rats were randomly assigned to three groups that underwent either fourth and fifth intercostal nerve ligation, cutting of the fourth and fifth ribs, or a sham operation in which only pleura was cut. For behavioural response assessment during the following month, pinch and touch were used as mechanical stimuli, and acetone was used as a cold thermal stimulus. In addition, (125)I-substance P autoradiography was used to determine neurokinin (NK) receptor density in spinal cord laminae I and II at one to six weeks after surgery. RESULTS: In rats with nerve ligation, hypersensitivity to noxious and non-noxious stimuli continued throughout the month. The "mirror phenomenon" was observed. The lowest threshold was obtained in the dorsomedial portion of the T4 dermatome on the side of surgery. In rats with rib cutting, a lowered threshold to noxious and non-noxious stimuli was observed for two weeks. In rats with sham operations, hypersensitivity was seen only at postoperative day one. NK-1 receptor density on the side of operation increased significantly in rats with nerve ligation from day seven to 28. Receptor density was highest on day 14 (22.97 +/- 1.04 fmol x mg(-1) tissue vs. control, 16.22 +/- 0.43), representing a 50% receptor excess on the side of ligation compared to the contralateral side. CONCLUSION: Intercostal nerve damage induces long-term postthoracotomy pain and an increase of spinal NK 1 receptors in rats. This model may be useful for investigation of postthoracotomy pain. PMID- 11495875 TI - Low dose intrathecal ropivacaine with or without sufentanil provides effective analgesia and does not impair motor strength during labour: a pilot study. AB - PURPOSE: Although ropivacaine has been used to provide spinal anesthesia in the surgical population, its intrathecal administration for labour analgesia has only recently been described. We evaluated the effects of low dose intrathecal ropivacaine with or without sufentanil for labour analgesia. METHODS: Thirty-six term parturients in active labour were randomly assigned to receive 3 mg of intrathecal ropivacaine (group R) or 3 mg ropivacaine with 10 microg of sufentanil (group RS). Patients were evaluated by a blinded observer for hypotension, linear analogue score (VAS 0-100) for labour pain, motor power in the lower limbs, onset of analgesia, sensation to cold and pin prick, duration of analgesia, and neonatal Apgar scores. The following day patients were assessed for satisfaction, headache and neurologic deficit. RESULTS: The mean duration of analgesia in the R group was 41.4 +/- 4.9 min and 95.0 +/- 6.1 min in the RS group (mean +/- SEM, P=0.0001). All subjects had satisfactory analgesia at five minutes, although analgesia from the ropivacaine- sufentanil combination was superior to that provided by ropivacaine alone. Total duration of labour was no different between the groups (R- 306 +/- 34, RS- 384 +/- 44 min, P=0.17). No patient showed evidence of motor block. All patients were satisfied with the labour analgesia. No neurological complications were observed. CONCLUSIONS: Low dose ropivacaine provides effective analgesia during labour via the intrathecal route. It can be mixed with sufentanil in the above-mentioned concentrations to improve both the quality and duration of analgesia. Fetal outcome remains favourable. It may provide minimal or no motor block, to facilitate ambulation. PMID- 11495876 TI - Combined spinal-epidural anesthesia for Cesarean section in a patient with peripartum dilated cardiomyopathy. AB - PURPOSE: To report a case of peripartum dilated cardiomyopathy associated with morbid obesity and possible difficult airway presenting for elective Cesarean section, which was successfully managed with combined spinal-epidural anesthesia. CLINICAL FEATURES: A morbidly obese parturient with a potentially difficult airway, suffering from idiopathic peripartum cardiomyopathy (ejection fraction 20%), was scheduled for an elective Cesarean section. A combined spinal epidural anesthesia was performed and 6 mg of bupivacaine were injected into the subarachnoid space. This was supplemented after 60 min with 25 mg of bupivacaine injected epidurally. The patient's hemodynamic status was monitored with direct intra-arterial blood pressure and central venous pressure measurements. The patient's perioperative course was uneventful. CONCLUSION: In patients suffering from peripartum cardiomyopathy, undergoing Cesarean section, combined spinal epidural anesthesia may be an acceptable anesthetic alternative. PMID- 11495878 TI - Confirmation of internal jugular guide wire position utilizing transesophageal echocardiography. AB - PURPOSE: To describe the utility of transesophageal echocardiography (TEE) in confirmation of correct central line J-wire position. CLINICAL FEATURES: A 51-yr old male patient presented for urgent coronary artery bypass grafting. Current medications included aspirin, nitroglycerine, heparin and a B-blocker. Physical examination was unremarkable. Initial difficulty with right internal jugular vein cannulation was encountered. A posterior approach was used to access the vein with further difficulty in passing the J-wire. Simultaneous TEE images confirmed the correct J-wire position. CONCLUSION: The case demonstrates the value of TEE to confirm correct guide wire position prior to insertion of a large bore central venous catheter. TEE visualization of J-wire position avoided repeat attempts at internal jugular cannulation and potential carotid artery puncture. PMID- 11495877 TI - Airway obstruction in a child with asymptomatic tracheobronchomalacia. AB - PURPOSE: To report a case of airway obstruction with hypoxia during emergence from anesthesia due to unexpected tracheobronchomalacia in a child. CLINICAL FEATURES: In a previously healthy 22-month-old boy with no symptoms or signs of respiratory disease, general anesthesia was induced by inhalation of increasing concentrations of sevoflurane (up to 5%) in oxygen and a laryngeal mask was inserted. Partial airway obstruction persisted during surgery, but obstruction was relieved by positive-pressure ventilation. During emergence from anesthesia, airway obstruction with hypoxia occurred, necessitating tracheal intubation. Emission of carbon dioxide as well as of sevoflurane was reduced and emergence from anesthesia markedly delayed. Fibreoptic tracheoscopy showed marked collapse of the tracheobronchi during expiration, and a diagnosis of tracheobronchomalacia was made. No respiratory complications occurred postoperatively. CONCLUSION: Asymptomatic tracheomalacia should also be suspected in case of airway obstruction during anesthesia in young children. PMID- 11495879 TI - Video-intuboscopic assistance is a useful aid to tracheal intubation in pediatric patients. AB - PURPOSE: To evaluate the efficacy of video-intuboscopic assisted tracheal intubation in a difficult intubation setting. METHODS: In 50 pediatric patients (mean age 12.8 +/- 3.1 yr, range 6-16 yr) a grade 3 direct laryngoscopic view was simulated. Eight certified registered nurse anesthetists without experience in endoscopic intubation performed tracheal intubation on five or more patients using the video-optical intubation stylet. Time from insertion of the tube into the oral cavity until the tip had passed the vocal cords was recorded. Failed intubation was defined as intubation >60 sec, arterial oxygen saturation <92% or esophageal intubation. Subjective degree of difficulty was asked from the operators using a Likert-scale. RESULTS: Forty-six of the 50 patients were successfully intubated within 60 sec and without arterial oxygen desaturation. In four patients, video-assisted tracheal intubation failed due to prolonged intubation time. Intubation times ranged from 10-40 sec (median 15 sec). Mean intubation time in the first patient (24.5 +/- 17.3 sec) appeared longer than for the fifth patient (20.8 +/- 10.9 sec), but the difference was not statistically significant (P=0.87). Mean estimated degree of difficulty was 3.9 +/- 2.1. Subjective estimates of difficulty increased with intubation times (P=0.001). CONCLUSION: The video-optical intubation stylet can be considered a valuable aid for tracheal intubation in pediatric patients with a difficult airway. PMID- 11495881 TI - Low frequency jet ventilation for stent insertion in a patient with tracheal stenosis. AB - PURPOSE: Evaluate oxygen jet ventilation in a patient with tracheal stenosis undergoing stent insertion. CLINICAL FEATURES: Manual intermittent low frequency oxygen jet ventilation was used during general anesthesia for fibreoptic bronchoscopy and stent insertion in a patient with tracheal stenosis. Oxygen jets were delivered via a Sander's injector adapted to the proximal end of the endotracheal tube on one side, and open to room air on the other side. Adequate oxygenation and carbon dioxide removal were ensured throughout the procedure. CONCLUSION: Low frequency jet ventilation in a patient with tracheal stenosis provided adequate ventilation as well as a non- obstructed field during fibreoptic bronchoscopy and stent insertion. PMID- 11495880 TI - Loss of the airway during tracheostomy: rescue oxygenation and re-establishment of the airway. AB - PURPOSE: To describe loss of the airway during tracheostomy and suggest a method for re-establishment of the airway and providing rescue oxygenation. CLINICAL FEATURES: A 22-yr-old female diagnosed with encephalomyelopathy was admitted to the intensive care unit with a progressively deteriorating level of consciousness and respiratory failure requiring intubation and ventilation. Several weeks later, an elective tracheostomy was performed under anesthesia. The surgeon made an anterior tracheal wall incision and inserted a cuffed #6 Shiley tracheostomy tube. No end-tidal CO(2) was detected and the patient could not be ventilated. After another failed attempt at insertion of a second tracheostomy tube, the diagnosis was made of a false passage within the trachea. The Shiley tracheostomy tube was removed and a #6 regular endotracheal tube was introduced in the trachea through the tracheostomy incision. The patient now could be ventilated with difficulty and low readings of end-tidal CO(2) were noted. Despite all efforts to further ventilate the patient, the arterial oxygen saturation never recovered, resulting in cardiac arrest. CONCLUSION: To restore a lost airway during tracheostomy, we recommend that a jet ventilation airway exchange catheter (JVAE) be inserted in the endotracheal tube through a bronchoscope port attachment prior to surgical entry into the trachea. The JVAE will also ensure continued ability to oxygenate the patient. PMID- 11495882 TI - Capnography confirms correct feeding tube placement in intensive care unit patients. AB - PURPOSE: To test the accuracy and potential time savings of capnography as compared with a two-step radiographic method in placing feeding tubes in critically ill patients. METHODS: One hundred feeding tube placements were studied in our tertiary care intensive care unit. All placements utilized a two step radiographic method, but capnography was added to the procedure. The procedure was then completed or abandoned depending on radiographic interpretation. RESULTS: Radiography showed 11 feeding tubes projecting within the tracheobronchial tree. In all 11 of these placements, the capnography unit displayed a normal capnogram. Radiography revealed 86 tube placements in the midesophageal region. In all 86 of these placements, capnography displayed a "purging warning". In three placements, radiography indicated that the tube was coiled in the oropharynx. In these cases, the capnograph displayed one "no purging/no capnogram" result, and two "purging" warnings. If using capnography alone, an average of 72.5 min would be required to complete a feeding tube placement (which includes time for requisite "pre-feed radiograph"). The two-step radiological approach took an average of 169.4 min, a difference of 96.9 min (P <0.0001) between the two methods. CONCLUSIONS: Capnography accurately identified all intratracheal feeding tube placements in this study. This study also shows that the use of capnography would significantly shorten the time needed for tube placement compared with a two-step radiologic method. Capnography should be considered for routine use when placing feeding tubes since it adds little time to the procedure and may improve patient safety. PMID- 11495884 TI - Wrist actigraphy in anesthesia. PMID- 11495885 TI - Syringe contamination by propofol: a possible mechanism. PMID- 11495886 TI - Opening an ampoule? Start from a scratch. PMID- 11495887 TI - A new setup for emergency transtracheal jet ventilation. PMID- 11495888 TI - Effectiveness of transnasal jet ventilation - a teaching aid. PMID- 11495889 TI - Central venous catheters for craniotomies. PMID- 11495890 TI - An aid to correct positioning of the ProSeal laryngeal mask. PMID- 11495892 TI - Prediction of difficult intubation: are we talking about the same thing? PMID- 11495891 TI - Intraoperative problem during surgery for Chiari malformation. PMID- 11495893 TI - Puncture of the laryngeal mask airway during stellate ganglion block. PMID- 11495894 TI - The multidrug resistance P-glycoprotein. Oligomeric state and intramolecular interactions. AB - The human multidrug resistance P-glycoprotein (P-gp), a member of the ATP-binding cassette (ABC) superfamily of transporters, is frequently responsible for the failure of chemotherapy by virtue of its ability to export hydrophobic cytotoxic drugs from cells. Elucidating the inter- and intramolecular interactions of this protein is critical to understanding its cellular function and mechanism of action. Toward this end, we have used both biochemical and genetic techniques to probe potential oligomerization interactions of P-gp. Differentially epitope tagged P-gp molecules did not co-immunoprecipitate when co-expressed in HEK293 cells or when co-translated in vitro, demonstrating that P-gp is monomeric in both the presence and absence of detergents. The two cytoplasmic domains of P-gp did not interact with each other in vivo when co-expressed as gene fusions in yeast. In contrast, the homologous domains of the transporter associated with antigen processing (TAP), which reside on separate polypeptides and must form a heterodimeric transporter (TAP1/TAP2), did interact in this system, suggesting a role for these domains in TAP dimerization. Implications for understanding the subunit organization of ABC transporters are discussed. PMID- 11495895 TI - Identification of a dominant negative mutant of Sprouty that potentiates fibroblast growth factor- but not epidermal growth factor-induced ERK activation. AB - Various mitogenic stimuli such as epidermal growth factor (EGF), fibroblast growth factor (FGF), and phorbol 12,13-dibutyrate (PDBu) activate the Ras-Raf-MEK ERK pathway, but the regulatory mechanism of this pathway remains to be investigated. Here we found that in 293 cells, mammalian Sprouty2 and Sprouty4 were rapidly induced by EGF, FGF, and PDBu in an ERK pathway-dependent manner. Forced expression of Sprouty2 and Sprouty4 inhibited FGF-induced ERK activation but did not affect EGF- or PDBu-induced ERK activation. To examine whether endogenous Sproutys were also selective inhibitors, we generated a dominant negative form of Sprouty2 (Y55A) and Sprouty4 (Y53A) in which conserved tyrosine residues were mutated. These mutants reverted the suppressive effect of both Sprouty2 and Sprouty4 but not that of RasGAP or SPRED (Sprouty-related EVH1 domain-containing protein), another Sprouty-related Ras suppressor. Expression of dominant negative Sprouty2 and Sprouty4 enhanced and prolonged FGF- but not EGF induced ERK activation in 293 cells. In PC12 cells, endogenous Sprouty4 was also induced by FGF. Overexpression of wild-type Sprouty4 blocked FGF-induced differentiation, whereas Y53A-Sprouty4 enhanced it. These observations suggest that endogenous Sprouty2 and Sprouty4 are physiological negative feedback regulators of growth factor-mediated ERK pathway and that there are Sprouty sensitive and -insensitive ERK activation pathways. Finding a dominant negative form of Sproutys will facilitate the study of the molecular mechanism and physiological function of Sproutys. PMID- 11495896 TI - Hemoglobin-degrading, aspartic proteases of blood-feeding parasites: substrate specificity revealed by homology models. AB - Blood-feeding parasites, including schistosomes, hookworms, and malaria parasites, employ aspartic proteases to make initial or early cleavages in ingested host hemoglobin. To better understand the substrate affinity of these aspartic proteases, sequences were aligned with and/or three-dimensional, molecular models were constructed of the cathepsin D-like aspartic proteases of schistosomes and hookworms and of plasmepsins of Plasmodium falciparum and Plasmodium vivax, using the structure of human cathepsin D bound to the inhibitor pepstatin as the template. The catalytic subsites S5 through S4' were determined for the modeled parasite proteases. Subsequently, the crystal structure of mouse renin complexed with the nonapeptidyl inhibitor t-butyl-CO-His-Pro-Phe-His-Leu [CHOHCH(2)]Leu-Tyr-Tyr-Ser- NH(2) (CH-66) was used to build homology models of the hemoglobin-degrading peptidases docked with a series of octapeptide substrates. The modeled octapeptides included representative sites in hemoglobin known to be cleaved by both Schistosoma japonicum cathepsin D and human cathepsin D, as well as sites cleaved by one but not the other of these enzymes. The peptidase-octapeptide substrate models revealed that differences in cleavage sites were generally attributable to the influence of a single amino acid change among the P5 to P4' residues that would either enhance or diminish the enzymatic affinity. The difference in cleavage sites appeared to be more profound than might be expected from sequence differences in the enzymes and hemoglobins. The findings support the notion that selective inhibitors of the hemoglobin-degrading peptidases of blood-feeding parasites at large could be developed as novel anti parasitic agents. PMID- 11495897 TI - Identification of protein kinase Calpha as an essential, but not sufficient, cytosolic factor for Ca2+-induced alpha- and dense-core granule secretion in platelets. AB - Upon activation, platelets release many active substances. Here, we have analyzed the mechanism governing Ca(2+)-induced secretion of von Willebrand factor stored in alpha-granules and 5-hydroxytryptamine in dense-core granules in permeabilized human platelets. Both secretions were dependent on ATP and cytosol. An essential factor for both granule secretions was purified from rat brain cytosol and identified to be protein kinase Calpha (PKCalpha) by partial amino acid sequencing. Purified PKCalpha efficiently stimulated both secretions in the presence of cytosol, whereas PKCalpha alone did not support the secretion of either type of granules, suggesting that PKCalpha is not a sufficient factor. Finally, in human platelet cytosol fractionated by a gel filtration column, the stimulatory activity for dense-core granule secretion paralleled with the concentration of PKC, suggesting that PKC could also be such a stimulatory factor in platelet cytosol. Thus, we identified PKCalpha as an essential, but not sufficient, cytosolic factor for the Ca(2+)-induced secretions of both alpha- and dense-core granules in platelets. PMID- 11495898 TI - Nerve growth factor inhibits apoptosis in memory B lymphocytes via inactivation of p38 MAPK, prevention of Bcl-2 phosphorylation, and cytochrome c release. AB - Survival of memory B lymphocytes is tightly linked to the integrity of the Bcl-2 protein and is regulated by a nerve growth factor (NGF) autocrine circuit. In factor-starved memory B cells, the addition of exogenous NGF promptly induced p38 mitogen-activated protein kinase (MAPK), but not c-Jun N-terminal kinase (JNK), dephosphorylation. Conversely, withdrawal of endogenous NGF was followed by p38 MAPK activation and translocation onto mitochondria, whereby it combined with and phosphorylated Bcl-2, as assessed by co-immunoprecipitation and kinase assays in vivo and in vitro. Mitochondria isolated from human memory B cells, then exposed to recombinant p38 MAPK, released cytochrome c, as did mitochondria from Bcl-2 negative MDCK cells loaded with recombinant Bcl-2. Apoptosis induced by NGF neutralization could be blocked by the specific p38 MAPK inhibitor SB203580 or by Bcl-2 mutations in Ser-87 or Thr-56. These data demonstrate that the molecular mechanisms underlying the survival factor function of NGF critically rely upon the continuous inactivation of p38 MAPK, a Bcl-2-modifying enzyme. PMID- 11495899 TI - The transmembrane heregulin precursor is functionally active. AB - A variety of eucaryotic polypeptide growth factors are synthesized as transmembrane precursors. Many of these precursors are released from plasma membranes by proteolytic cleavage and converted into soluble mature proteins. A number of studies, however, indicate that bound growth factor precursors can be biologically active, suggesting a role for these membrane-associated ligands in cell-cell communication. Secreted heregulin is a 45-kDa growth factor with homology to epidermal growth factor. This growth factor binds directly to HER-3 and HER-4 and activates heterodimeric receptor complexes composed of the type I receptor tyrosine kinases, i.e. HER-1, HER-2, HER-3, and HER-4. Heregulin was originally detected in the conditioned medium of the human breast cancer cell line MDA-MB-231 and purified based on its ability to stimulate phosphorylation of p185(HER-2/neu). In the current study, the biologic activity of plasma membrane anchored heregulin was evaluated in human breast cells. Transmembrane heregulin binds to cells expressing p180(HER-3), induces p185(HER-2/neu) phosphorylation, and increases DNA synthesis in cells overexpressing the HER-2/neu gene product. In addition, when cells containing heregulin receptors are co-cultured with heregulin-producing cells, specific in vivo associations are observed. This study demonstrates that transmembrane heregulin is functionally active and suggest it is capable of playing a role in cell-cell communication and subsequent signal transduction in vivo. PMID- 11495900 TI - Identification of residues of the Saccharomyces cerevisiae G protein-coupled receptor contributing to alpha-factor pheromone binding. AB - The Saccharomyces cerevisiae pheromone, alpha-factor (WHWLQLKPGQPMY), and Ste2p, its G protein-coupled receptor, were studied as a model for peptide ligand receptor interaction. The affinities and activities of various synthetic position 10 alpha-factor analogs with Ste2p expressing mutations at residues Ser47 and Thr48 were investigated. All mutant receptors were expressed at a similar level in the cytoplasmic membrane, and their efficacies of signal transduction were similar to that of the wild-type receptor. Mutant receptors differed in binding affinity (Kd) and potency (EC50) for gene induction by alpha-factor. One mutant receptor (S47K,T48K) had dramatically reduced affinity and activity for [Lys10]- and [Orn10]alpha-factor, whereas the affinity for Saccharomyces kluyveri alpha factor (WHWLSFSKGEPMY) was increased over 20-fold compared with that of wild-type receptor. In contrast, the affinity of [Lys10]- and [Orn10]alpha-factor was increased greatly in a S47E,T48E mutant receptor, whereas the binding of the S. kluyveri alpha-factor was abolished. The affinity of [Lys10]- and [Orn10]alpha factor for the S47E,T48E receptor dropped 4-6-fold in the presence of 1 m NaCl, whereas the affinity of alpha-factor was not affected by this treatment. These results demonstrate that when bound to its receptor the 10th residue (Gln) of the S. cerevisiae alpha-factor is adjacent to Ser47 and Thr48 residues in the receptor and that the 10th residue of alpha-factors from two Saccharomyces species is responsible for the ligand selectivity to their cognate receptors. Based on these data, we have developed a two-dimensional model of alpha-factor binding to its receptor. PMID- 11495901 TI - PTEN induces chemosensitivity in PTEN-mutated prostate cancer cells by suppression of Bcl-2 expression. AB - The tumor suppressor gene PTEN (MMAC1/TEP1) is lost frequently in advanced prostate cancer (PCa). However, the function of PTEN in tumorigenesis is not understood fully. In this study, we demonstrate that expression of Bcl-2 in prostate tumors correlates with loss of the PTEN protein. This finding was verified by studies in the PCa cell lines DU145, PC-3, LNCaP, and an androgen refractory subline of LNCaP. Transient transfection of PTEN into the PTEN-null cells resulted in decreased levels of Bcl-2 mRNA and protein. These effects appear to be mediated at the level of gene transcription, since a Bcl-2 promoter reporter construct was down-regulated by ectopic expression of PTEN in LNCaP cells. The inhibition of Bcl-2 required the lipid-phosphatase activity of PTEN and was blocked by overexpression of a constitutively active form of Akt. Moreover, the transcription-regulatory protein cAMP-response element-binding protein (CREB) may be involved, since decreased phosphorylation of CREB at Ser(133) was detected following PTEN expression, and ectopic expression of CREB repressed completely the PTEN-induced inhibition of Bcl-2 promoter activity. Furthermore, cotransfection of Bcl-2 and PTEN expression vectors rescued PTEN induced cell death but not G(1) cell cycle arrest. Finally, forced expression of PTEN sensitized LNCaP cells to cell death induced by staurosporine, doxorubicin, and vincristine, and this chemosensitivity was attenuated by exogenous expression of Bcl-2. Taken together, these data demonstrate that loss of PTEN leads to up regulation of the bcl-2 gene, thus contributing to survival and chemoresistance of PCa cells. These findings suggest that the PTEN gene and its regulated pathway are potential therapeutic targets in prostate cancer. PMID- 11495902 TI - Alteration of cardiac and renal functions in transgenic mice overexpressing human mineralocorticoid receptor. AB - The mineralocorticoid receptor (MR), a ligand-dependent transcription factor, mediates aldosterone actions in a large variety of tissues. To explore the functional implication of MR in pathophysiology, transgenic mouse models were generated using the proximal human MR (hMR) promoter to drive expression of hMR in aldosterone target tissues. Tissue-specific analysis of transgene expression in two independent transgenic animal (TG) lines by ribonuclease protection assays revealed that hMR is expressed in all mineralocorticoid-sensitive tissues, most notably in the kidney and the heart. TG exhibit both renal and cardiac abnormalities. Enlarged kidneys were histologically associated with renal tubular dilation and cellular vacuolization whose prevalence increased with aging. Renal clearance studies also disclosed a significant decrease in urinary potassium excretion rate in TG. hMR-expressing animals had normal blood pressure but developed mild dilated cardiomyopathy (increased left ventricle diameters and decreased shortening fraction), which was accompanied by a significant increase in heart rate. Differential gene expression analysis revealed a 2- to 5-fold increase in cardiac expression of atrial natriuretic peptide, serum- and glucocorticoid-induced kinase, and early growth response gene 1 as detected by microarrays; renal serum- and glucocorticoid-induced kinase was also induced significantly. Altogether, TG exhibited specific alteration of renal and cardiac functions, thus providing useful pathophysiological models to gain new insights into the tissue-specific mineralocorticoid signaling pathways. PMID- 11495903 TI - BH3-only Bcl-2 family members are coordinately regulated by the JNK pathway and require Bax to induce apoptosis in neurons. AB - The Bcl-2 family of proteins are key regulators of programmed cell death. A distinct subfamily of BH3-only molecules has been identified, but their exact mechanism of action remains unclear. Here we show that the BH3-only Bcl-2 family members, Dp5/Hrk and Bim, are induced upstream of the Bax checkpoint in neuronal apoptosis in a manner that shows significant dependence on JNK signaling. We also show that Dp5 and other BH3-only proteins kill cerebellar granule neurons in a Bax-dependent manner. These studies demonstrate that BH3-only members do not act independently in their proapoptotic activities but rather require the action of multidomain proapoptotic Bcl-2 family members to produce cell death. PMID- 11495904 TI - alpha4beta3delta GABA(A) receptors characterized by fluorescence resonance energy transfer-derived measurements of membrane potential. AB - Selective modulators of gamma-aminobutyric acid, type A (GABA(A)) receptors containing alpha(4) subunits may provide new treatments for epilepsy and premenstrual syndrome. Using mouse L(-tk) cells, we stably expressed the native GABA(A) receptor subunit combinations alpha(3)beta(3)gamma(2,) alpha(4)beta(3)gamma(2), and, for the first time, alpha(4)beta(3)delta and characterized their properties using a novel fluorescence resonance energy transfer assay of GABA-evoked depolarizations. GABA evoked concentration dependent decreases in fluorescence resonance energy transfer that were blocked by GABA(A) receptor antagonists and, for alpha(3)beta(3)gamma(2) and alpha(4)beta(3)gamma(2) receptors, modulated by benzodiazepines with the expected subtype specificity. When combined with alpha(4) and beta(3), delta subunits, compared with gamma(2), conferred greater sensitivity to the agonists GABA, 4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridin-3-ol (THIP), and muscimol and greater maximal efficacy to THIP. alpha(4)beta(3)delta responses were markedly modulated by steroids and anesthetics. Alphaxalone, pentobarbital, and pregnanolone were all 3-7-fold more efficacious at alpha(4)beta(3)delta compared with alpha(4)beta(3)gamma(2.) The fluorescence technique used in this study has proven valuable for extensive characterization of a novel GABA(A) receptor. For GABA(A) receptors containing alpha(4) subunits, our experiments reveal that inclusion of delta instead of gamma(2) subunits can increase the affinity and in some cases the efficacy of agonists and can increase the efficacy of allosteric modulators. Pregnanolone was a particularly efficacious modulator of alpha(4)beta(3)delta receptors, consistent with a central role for this subunit combination in premenstrual syndrome. PMID- 11495905 TI - Differential internalization of mammalian and non-mammalian gonadotropin releasing hormone receptors. Uncoupling of dynamin-dependent internalization from mitogen-activated protein kinase signaling. AB - Desensitization and internalization of G-protein-coupled receptors can reflect receptor phosphorylation-dependent binding of beta-arrestin, which prevents G protein activation and targets receptors for internalization via clathrin-coated vesicles. These can be pinched off by a dynamin collar, and proteins controlling receptor internalization can also mediate mitogen-activated protein kinase signaling. Gonadotropin-releasing hormone (GnRH) stimulates internalization of its receptors via clathrin-coated vesicles. Mammalian GnRH receptors (GnRH-Rs) are unique in that they lack C-terminal tails and do not rapidly desensitize, whereas non-mammalian GnRH-R have C-terminal tails and, where investigated, do rapidly desensitize and internalize. Using recombinant adenovirus expressing human and Xenopus GnRH-Rs we have explored the relationship between receptor internalization and mitogen-activated protein kinase signaling in HeLa cells with regulated tetracycline-controlled expression of wild-type or a dominant negative mutant (K44A) of dynamin. These receptors were phospholipase C-coupled and had appropriate ligand affinity and specificity. K44A dynamin expression did not alter human GnRH-R internalization but dramatically reduced internalization of Xenopus GnRH-R (and epidermal growth factor (EGF) receptor). Blockade of clathrin mediated internalization (sucrose) abolished internalization of all three receptors. Both GnRH-Rs also mediated phosphorylation of ERK 2 and for both receptors, this was inhibited by K44A dynamin. The same was true for EGF- and protein kinase C-mediated ERK 2 phosphorylation. ERK 2 phosphorylation was also inhibited by a protein kinase C inhibitor but not affected by an EGF receptor tyrosine kinase inhibitor. We conclude that a) desensitizing and non desensitizing GnRH-Rs are targeted for clathrin-coated vesicle-mediated internalization by functionally distinct mechanisms, b) GnRH-R signaling to ERK 2 is dynamin-dependent and c) this does not reflect a dependence on dynamin dependent GnRH-R internalization. PMID- 11495906 TI - The role and predicted propensity of conserved proline residues in the 5-HT3 receptor. AB - 5-HT3 receptors possess a number of highly conserved proline residues. We changed each of these to alanine, expressed the mutants as homomeric 5-HT3A receptors in HEK293 cells, and analyzed them with radioligand binding, electrophysiology, and immunocytochemistry. Mutation of Pro56, Pro104, Pro123, and Pro170 resulted in ablation of radioligand binding, whereas mutation of Pro257 and Pro301 did not. Only the latter were expressed at the plasma membrane but were non-functional. Thus the former, which are in the N-terminal domain, may be involved in forming correct receptor structure, while those in the transmembrane region (Pro257 and Pro301) are necessary for the function of the protein. To explore the conformational preference (propensity) of these residues we examined the proportion of cis-prolines and the influence of adjacent residues in known protein structures. 4.7% of prolines in the protein data base were in the cis conformation, and the distribution of amino acids adjacent to cis-prolines was not randomly distributed. Comparison of the proportion of each amino acid residue adjacent to a cis-proline revealed that aromatic and bend-facilitating residues were favored while those with beta-branched chains were not. Thus five residues (Gly, Pro, Tyr, Trp, Phe) and three residues (Pro, Tyr, Phe) were found more frequently than expected before and after cis-prolines respectively, whereas five residues (Val, Ile, Leu, Asp, Thr) and two residues (Asp, Glu) were found less frequently. Of the 20 proline residues in the 5-HT3A receptor subunit only Pro170 has adjacent residues that are favorable. Mutating these to non-favorable residues resulted in ablation of ligand binding, whereas replacement with alternative favorable residues did not. We therefore propose that Pro170, which is part of the characteristic cys-loop found in this family of proteins, may be in the cis conformation. PMID- 11495907 TI - Subfilamentous protofibril structures in fibrous proteins: cross-linking evidence for protofibrils in intermediate filaments. AB - The packing of the constituent molecules in some fibrous proteins such as collagen and intermediate filaments (IF) is thought to consist of several hierarchical levels, the penultimate of which is the organization of subfilamentous units termed protofibrils. However, to date only indirect evidence, such as electron microscopic images of unraveling fibers or the existence of mass quanta, has been adduced in support of the existence of protofibrils. We have reexamined this issue in IF. Cross-links have been induced in trichocyte keratin, cytokeratin, and vimentin IF proteins. Using improved experimental conditions, several additional and reproducible cross-links have been characterized. Notably, many of these link between columns of molecular strands four apart on two-dimensional surface lattices. These data provide robust support for the concept of an 8-chain (4-molecule) protofibril entity in IF. Further, their positions correspond to the axial displacements predicted for protofibrils in the different types of IF. Also, the data are consistent with intact IF containing four protofibrils. In addition, the positions of these novel cross-links suggest that there are multiple possible groupings of four molecular strands to form a protofibril, suggesting a promiscuous association of molecules to form a protofibril. This may underlie the reason that organized elongated protofibrils cannot be visualized by conventional microscopic methods. PMID- 11495908 TI - Identification and characterization of a family of Rab11-interacting proteins. AB - Rab11a is a small GTP-binding protein enriched in the pericentriolar plasma membrane recycling systems. We hypothesized that Rab11a-binding proteins exist as downstream effectors of its action. Here we define a family of four Rab11 interacting proteins: Rab11-Family Interacting Protein 1 (Rab11-FIP1), Rab11 Family Interacting Protein 2 (Rab11-FIP2), Rab11-Family Interacting Protein 3 (Rab11-FIP3), and pp75/Rip11. All four interacting proteins associated with wild type Rab11a and dominant active Rab11a (Rab11aS20V) as well as Rab11b and Rab25. Rab11-FIP2 also interacted with dominant negative Rab11a (Rab11aS25N) and the tail of myosin Vb. The binding of Rab11-FIP1, Rab11-FIP2, and Rab11-FIP3 to Rab11a was dependent upon a conserved carboxyl-terminal amphipathic alpha-helix. Rab11-FIP1, Rab11-FIP2, and pp75/Rip11 colocalized with Rab11a in plasma membrane recycling systems in both non-polarized HeLa cells and polarized Madin-Darby canine kidney cells. GFP-Rab11-FIP3 also colocalized with Rab11a in HeLa cells. Rab11-FIP1, Rab11-FIP2, and pp75/Rip11 also coenriched with Rab11a and H(+)K(+) ATPase on parietal cell tubulovesicles, and Rab11-FIP1 and Rab11-FIP2 translocated with Rab11a and the H(+)K(+)-ATPase upon stimulating parietal cells with histamine. The results suggest that the function of Rab11a in plasma membrane recycling systems is dependent upon a compendium of protein effectors. PMID- 11495909 TI - Characterization of receptors of insect cytokine, growth-blocking peptide, in human keratinocyte and insect Sf9 cells. AB - Insect cytokine, growth-blocking peptide (GBP), enhances cell proliferation of human keratinocyte cells with a potency almost equivalent to that of human epidermal growth factor (EGF). GBP consists of 25 amino acid residues containing a core region that shows a striking similarity to the C-terminal beta-loop domain of EGF and disordered N and C termini. The present study demonstrates that, although GBP lacks the N-terminal half-portion of EGF molecule, at least five amino acids of the disordered N-terminal six-amino acid region are indispensable for affecting the cell growth activity of GBP. Upon stimulating mitogenesis in keratinocyte cells, GBP directly binds and activates their EGF receptors. GBP also effects proliferative activity on insect Sf9 cells through the binding and activation of the specific receptor, which consists of a heterodimeric complex: a binding subunit (60 kDa) and a tyrosine phosphorylation subunit (58 kDa). These results indicate that GBP enhances cell proliferation of human keratinocyte and insect Sf9 cells through the activation of EGF and GBP receptors, respectively. PMID- 11495910 TI - Control of conformational equilibria in the human B2 bradykinin receptor. Modeling of nonpeptidic ligand action and comparison to the rhodopsin structure. AB - A prototypic study of the molecular mechanisms of activation or inactivation of peptide hormone G protein-coupled receptors was carried out on the human B2 bradykinin receptor. A detailed pharmacological analysis of receptor mutants possessing either increased constitutive activity or impaired activation or ligand recognition allowed us to propose key residues participating in intramolecular interaction networks stabilizing receptor inactive or active conformations: Asn(113) and Tyr(115) (TM III), Trp(256) and Phe(259) (TM VI), Tyr(295) (TM VII) which are homologous of the rhodopsin residues Gly(120), Glu(122), Trp(265), Tyr(268), and Lys(296), respectively. An essential experimental finding was the spatial proximity between Asn(113), which is the cornerstone of inactive conformations, and Trp(256) which plays a subtle role in controlling the balance between active and inactive conformations. Molecular modeling and mutagenesis data showed that Trp(256) and Tyr(295) constitute, together with Gln(288), receptor contact points with original nonpeptidic ligands. It provided an explanation for the ligand inverse agonist behavior on the WT receptor, with underlying restricted motions of TMs III, VI, and VII, and its agonist behavior on the Ala(113) and Phe(256) constitutively activated mutants. These data on the B2 receptor emphasize that conformational equilibria are controlled in a coordinated fashion by key residues which are located at strategic positions for several G protein-coupled receptors. They are discussed in comparison with the recently determined rhodopsin crystallographic structure. PMID- 11495911 TI - Calmodulin regulates assembly and trafficking of SK4/IK1 Ca2+-activated K+ channels. AB - Calmodulin (CaM) regulates gating of several types of ion channels but has not been implicated in channel assembly or trafficking. For the SK4/IK1 K+ channel, CaM bound to the proximal C terminus ("Ct1 " domain) acts as the Ca2+ sensor. We now show that CaM interacting with the C terminus of SK4 also controls channel assembly and surface expression. In transfected cells, removing free CaM by overexpressing the CaM-binding domain, Ct1, redistributed full-length SK4 protein from the plasma membrane to the cytoplasm and decreased whole-cell currents. Making more CaM protein available by overexpressing the CaM gene abrogated the dominant-negative effect of Ct1 and restored both surface expression of SK4 protein and whole-cell currents. The distal C-terminal domain ("Ct2") also plays a role in assembly, but is not CaM-dependent. Co-immunoprecipitation experiments demonstrated that multimerization of SK4 subunits was enhanced by CaM and inhibited by removal of CaM, indicating that CaM regulates trafficking of SK4 by affecting the assembly of channels. Our results support a model in which CaM dependent association of SK4 monomers at their Ct1 domains regulates channel assembly and surface expression. This appears to represent a novel mechanism for controlling ion channels, and consequently, the cellular functions that depend on them. PMID- 11495912 TI - Involvement of phosphatidylinositol 3-kinase and mitogen-activated protein kinases in glycine-extended gastrin-induced dissociation and migration of gastric epithelial cells. AB - The various molecular forms of gastrin can act as promoters of proliferation and differentiation in different regions of the gastrointestinal tract. We report a novel stimulatory effect of glycine-extended gastrin(17) only on cell/cell dissociation and cell migration in a non-tumorigenic mouse gastric epithelial cell line (IMGE-5). In contrast, both amidated and glycine-extended gastrin(17) stimulated proliferation of IMGE-5 cells via distinct receptors. Glycine-extended gastrin(17)-induced dissociation preceded migration and was blocked by selective inhibitors of phosphatidylinositol 3-kinase (PI3-kinase) but did not require mitogen-activated protein (MAP) kinase activation. Furthermore, glycine-extended gastrin(17) induced a PI3-kinase-mediated tyrosine phosphorylation of the adherens junction protein beta-catenin, partial dissociation of the complex between beta-catenin and the transmembrane protein E-cadherin, and delocalization of beta-catenin into the cytoplasm. Long lasting activation of MAP kinases by glycine-extended gastrin(17) was specifically required for the migratory response, in contrast to the involvement of a rapid and transient MAP kinase activation in the proliferative response to both amidated and glycine-extended gastrin(17). Therefore, the time course of MAP kinase activation appears to be a critical determinant of the biological effects mediated by this pathway. Together with the involvement of PI3-kinase in the dissociation of adherens junctions, long term activation of MAP kinases seems responsible for the selectivity of this novel effect of G(17)-Gly on the adhesion and migration of gastric epithelial cells. PMID- 11495913 TI - Osmotic shock induces G1 arrest through p53 phosphorylation at Ser33 by activated p38MAPK without phosphorylation at Ser15 and Ser20. AB - Osmotic shock induced transient stabilization of p53, possibly due to increased degradation of Mdm2. Stabilized p53 was activated by p38(MAPK), resulting in G(1) arrest through induction of p21(WAF1). Among the postulated phosphorylation sites involved in p53 stabilization or activation (Ser(15), Ser(20), Ser(33), and Ser(46)), only Ser(33) was phosphorylated. Furthermore, interaction of p53 with the transcriptional coactivator p300 was induced, and Lys(382) of p53 was acetylated. Although inhibition of p38(MAPK) did not prevent nuclear accumulation of p53, phosphorylation of Ser(33) was markedly suppressed by SB203580, a specific inhibitor of p38(MAPK). Under these conditions, acetylation of Lys(382) and induction of p21(WAF1) were also inhibited, and cells with elevated levels of p53 showed normal cell cycle progression. Activated p38(MAPK) phosphorylated endogenous p53 at Ser(33) in living cells. In stable transformants expressing dominant negative MKK6, an upstream protein kinase of p38(MAPK), p53 stabilization was induced normally following osmotic shock, but phosphorylation of Ser(33), acetylation of Lys(382), and induction of p21(WAF1) were almost completely inhibited. These results suggest that phosphorylation at Ser(33) by p38(MAPK) is critical for activation of p53 following osmotic shock. Phosphorylation of neither Ser(15) nor Ser(20) was needed in this activation. PMID- 11495914 TI - Interaction of tau isoforms with Alzheimer's disease abnormally hyperphosphorylated tau and in vitro phosphorylation into the disease-like protein. AB - The microtubule-associated protein tau is a family of six isoforms that becomes abnormally hyperphosphorylated and accumulates in neurons undergoing neurodegeneration in the brains of patients with Alzheimer disease (AD). We investigated the isoform-specific interaction of normal tau with AD hyperphosphorylated tau (AD P-tau). We found that the binding of AD P-tau to normal human recombinant tau was tau4L > tau4S > tau4 and tau3L > tau3S > tau3, and that its binding to tau4L was greater than to tau3L. AD P-tau also inhibited the assembly of microtubules promoted by each tau isoform and caused disassembly when added to preassembled microtubules. This inhibition and depolymerization of microtubules by the AD P-tau corresponded directly to the degree of its interaction with the different tau isoforms. In vitro hyperphosphorylation of recombinant tau (P-tau) conferred AD P-tau-like characteristics. Like AD P-tau, P tau interacted with and sequestered normal tau and inhibited microtubule assembly. These studies suggest that the AD P-tau interacts preferentially with the tau isoforms that have the amino-terminal inserts and four microtubule binding domain repeats and that hyperphosphorylation of tau appears to be sufficient to acquire AD P-tau characteristics. Thus, lack of amino-terminal inserts and extra microtubule binding domain repeat in fetal human brain might be protective from Alzheimer's neurofibrillary degeneration. PMID- 11495915 TI - Complexation of two proteic insect inhibitors to the active site of chymotrypsin suggests decoupled roles for binding and selectivity. AB - The crystal structures of two homologous inhibitors (PMP-C and PMP-D2v) from the insect Locusta migratoria have been determined in complex with bovine alpha chymotrypsin at 2.1- and 3.0-A resolution, respectively. PMP-C is a potent bovine alpha-chymotrypsin inhibitor whereas native PMP-D2 is a weak inhibitor of bovine trypsin. One unique mutation at the P1 position converts PMP-D2 into a potent bovine alpha-chymotrypsin inhibitor. The two peptides have a similar overall conformation, which consists of a triple-stranded antiparallel beta-sheet connected by three disulfide bridges, thus defining a novel family of serine protease inhibitors. They have in common the protease interaction site, which is composed of the classical protease binding loop (position P5 to P'4, corresponding to residues 26-34) and of an internal segment (residues 15-18), held together by two disulfide bridges. Structural divergences between the two inhibitors result in an additional interaction site between PMP-D2v (position P10 to P6, residues 21-25) and the residues 172-175 of alpha-chymotrypsin. This unusual interaction may be responsible for species selectivity. A careful comparison of data on bound and free inhibitors (from this study and previous NMR studies, respectively) suggests that complexation to the protease stabilizes the flexible binding loop (from P5 to P'4). PMID- 11495916 TI - Proapoptotic stimuli induce nuclear accumulation of glycogen synthase kinase-3 beta. AB - The goal of this study was to determine whether the intracellular distribution of the proapoptotic enzyme glycogen synthase kinase-3 beta (GSK-3 beta) is dynamically regulated by conditions that activate apoptotic signaling cascades. In untreated human neuroblastoma SH-SY5Y cells, GSK-3 beta was predominantly cytosolic, although a low level was also detected in the nucleus. The nuclear level of GSK-3 beta was rapidly increased after exposure of cells to serum-free media, heat shock, or staurosporine. Although each of these conditions caused changes in the serine 9 and/or tyrosine phosphorylation of GSK-3 beta, neither of these modifications was correlated with nuclear accumulation of GSK-3 beta. Heat shock and staurosporine treatments increased nuclear GSK-3 beta prior to activation of caspase-9 and caspase-3, and this nuclear accumulation of GSK-3 beta was unaltered by pretreatment with a general caspase inhibitor. The GSK-3 beta inhibitor lithium did not alter heat shock-induced nuclear accumulation of GSK-3 beta but increased the nuclear level of cyclin D1, indicating that cyclin D1 is a substrate of nuclear GSK-3 beta. Thus, the intracellular distribution of GSK-3 beta is dynamically regulated by signaling cascades, and apoptotic stimuli cause increased nuclear levels of GSK-3 beta, which facilitates interactions with nuclear substrates. PMID- 11495917 TI - The fused protein kinase regulates Hedgehog-stimulated transcriptional activation in Drosophila Schneider 2 cells. AB - The Drosophila segment polarity gene fused encodes a putative protein serine/threonine kinase, and plays a critical role in the signal transduction for Hedgehog (Hh)-dependent gene expression. We show that the Drosophila Schneider 2 (S2) cell line has the potential to transduce the Hh-triggered intracellular signals, leading to the activation of target gene expression, when a transcription factor, Cubitus interruptus (Ci), is provided exogenously. Using S2 cells transfected with the Ci-expressing plasmid and a patched promoter reporter construct, we demonstrate that the forced expression of Fused (Fu) stimulates Hh triggered and Ci-dependent transcriptional activation. The N-terminal kinase domain of Fu is required for this activity, but the C-terminal domain is not. Two kinase-inactive Fu mutants fail to enhance the reporter activation, indicating that the kinase catalytic activity is essential for this function. Negative components of the Hh-signaling pathway, Costal-2 and Suppressor of Fused, strongly antagonize the Fu activity, irrespective of the presence or absence of the Fu C-terminal domain, suggesting an indirect mechanism for the inhibition of Fu by these proteins. Furthermore, mutational analyses of threonine 158 and serine 159, in the activation segment of the Fu protein kinase, indicate that threonine 158 is essential for Fu activity and that phosphorylation of this threonine residue may be involved in the activation of the kinase catalytic activity upon Hh stimulation. PMID- 11495918 TI - Activity-dependent development of P2X7 current and Ca2+ entry in rabbit osteoclasts. AB - Bone remodeling is regulated by local factors and modulated by mechanical stimuli. Mechanical stimulation can cause release of ATP, an agent that stimulates osteoclastic resorption at low concentrations and inhibits at high concentrations. We examined whether osteoclasts express P2X(7) receptors, which are activated by high concentrations of ATP and can behave as ion channels or cause the formation of membrane pores. Rabbit osteoclasts were studied using patch clamp techniques. Successive or prolonged applications of 2'- & 3'-O-(4 benzoylbenzoyl)-ATP (BzATP, a relatively potent P2X(7) agonist) or high concentrations of ATP caused the development of a slowly deactivating inward current. The underlying channel was permeable only to small cations, ruling out pore formation. Divalent cations reduced current magnitude, consistent with the presence of P2X(7) receptors, a finding confirmed in rat osteoclasts by immunocytochemistry. Successive applications of BzATP also elicited [Ca(2+)](i) elevations that required extracellular Ca(2+). The BzATP-induced current and the rise of [Ca(2+)](i) were temporally associated, and both were inhibited by PPADS, a P2X(7) antagonist. This study demonstrates that high concentrations of ATP activate P2X(7) receptors and provides the first functional evidence for an extracellular ligand-gated Ca(2+) influx pathway in osteoclasts. ATP released in response to mechanical stimuli may act through P2X(7) receptors to inhibit osteoclastic resorption. PMID- 11495919 TI - Apoptosis signal-regulating kinase 1 controls the proapoptotic function of death associated protein (Daxx) in the cytoplasm. AB - Although Daxx (death-associated protein) was first reported to mediate the apoptotic signal from Fas to JNK in the cytoplasm, other data suggested that Daxx is mainly located in the nucleus as a transcriptional regulator. Here, we demonstrated that cellular localization of Daxx could be determined by the relative concentration of a proapoptotic kinase, apoptosis signal-regulating kinase 1 (ASK1) by using immunofluorescence and transcriptional reporter assay. ASK1 sequestered Daxx in the cytoplasm and inhibited the repressive activity of Daxx in transcription. In addition, Daxx was bound to the activated Fas only in the presence of ASK1, accelerating the Fas-mediated apoptosis. These results suggest that Daxx requires ASK1 for its cytoplasmic localization and Fas-mediated signaling. Taken together, we could conclude that ASK1 controls the dual function of Daxx as a transcriptional repressor in the nucleus and as a proapoptotic signal mediator in the cytoplasm. PMID- 11495920 TI - Novel transcriptional regulation of the human CYP3A7 gene by Sp1 and Sp3 through nuclear factor kappa B-like element. AB - Human CYP3A7 and CYP3A4 are expressed in fetal and adult livers, respectively, although the 5'-flanking regions of the two genes show 90% homology. The purpose of this study was to clarify the mechanism(s) responsible for the transcriptional regulation of the CYP3A7 gene in human hepatoma HepG2 cells that showed fetal phenotypes. Transfection studies using a series of the CYP3A7 or CYP3A4 promoter luciferase chimeric genes identified a nuclear factor kappaB (NF-kappaB)-like element between nucleotides -2326 and -2297 that conferred the transcriptional activation of the CYP3A7 gene. A 1-base pair mismatch within the corresponding region of the CYP3A4 gene was sufficient for a differential enhancer activity. A gel shift assay using nuclear extracts from HepG2 cells showed that Sp1 and Sp3 bound to the NF-kappaB-like element of the CYP3A7 but not CYP3A4 gene. Specific activation of the CYP3A7 promoter by Sp1 and Sp3 was confirmed by a co transfection of the p3A7NF-kappaB or p3A4NF-kappaB reporter gene with Sp1 or Sp3 expression plasmid into Drosophila cells, which lacked endogenous Sp family. Additionally, introduction of mutations into binding sites for hepatocyte nuclear factor 3beta, upstream stimulatory factor 1, and a basic transcription element in the proximal promoter attenuated luciferase activity to 20% of the level seen with the intact CYP3A7 promoter. Thus, we conclude that the expression of the CYP3A7 gene in HepG2 cells is cooperatively regulated by Sp1, Sp3, hepatocyte nuclear factor 3beta, and upstream stimulatory factor 1. PMID- 11495921 TI - Overexpression of beta 1 and beta 2 adrenergic receptors in rat atrial myocytes. Differential coupling to G protein-gated inward rectifier K(+) channels via G(s) and G(i)/o. AB - G protein-activated inwardly rectifying K(+) (GIRK) channels, expressed in atrial myocytes, various neurons, and endocrine cells, represent the paradigmatic target of beta gamma subunits released from activated heterotrimeric G proteins. These channels contribute to physiological slowing of cardiac frequency and synaptic inhibition. They are activated by beta gamma dimers released upon stimulation of receptors coupled to pertussis toxin-sensitive G proteins (G(i/o)), whereas beta gamma released from G(s) do not converge on the channel subunits. This is in conflict with the finding that dimeric combinations of various beta and gamma subunits can activate GIRK channels with little specificity. In the present study, we have overexpressed the major subtypes of cardiac beta-adrenergic receptors (beta(1)-AR and beta(2)-AR) in atrial myocytes by transient transfection. Whereas in native cells beta-adrenergic stimulation with isoproterenol failed to induce measurable GIRK current, robust currents were recorded from myocytes overexpressing either beta(1)-AR or beta(2)-AR. Whereas the beta(2)-AR-induced current showed the same sensitivity to pertussis toxin as the current evoked by the endogenous G(i/o)-coupled muscarinic M(2) receptor, isoproterenol-activated currents were insensitive to pertussis toxin treatment in beta(1)-AR-overexpressing myocytes. In contrast to a recent publication (Leaney, J. L., Milligan, G., and Tinker, A. (2000) J. Biol. Chem. 275, 921-929), sizable GIRK currents could also be activated by isoproterenol when the signaling pathway was reconstituted by transient transfection in two different standard cell lines (Chinese hamster ovary and HEK293). These results demonstrate that specificity of receptor-G protein signaling can be disrupted by overexpression of receptors. Moreover, the alpha subunit of heterotrimeric G proteins does not confer specificity to G beta gamma-mediated signaling. PMID- 11495922 TI - The biotinyl domain of Escherichia coli acetyl-CoA carboxylase. Evidence that the "thumb" structure id essential and that the domain functions as a dimer. AB - Biotin carboxyl carrier protein (BCCP) is the small biotinylated subunit of Escherichia coli acetyl-CoA carboxylase (ACC), the enzyme that catalyzes the first committed step of fatty acid synthesis. Similar proteins are found in other bacteria and in chloroplasts. E. coli BCCP is a member of a large family of protein domains modified by covalent attachment of biotin to a specific lysine residue. However, the BCCP biotinyl domain differs from many of these proteins in that an eight-amino acid residue insertion is present upstream of the biotinylated lysine. X-ray crystallographic and multidimensional NMR studies show that these residues constitute a structure that has the appearance of an extended thumb that protrudes from the otherwise highly symmetrical domain structure. I report that expression of two mutant BCCPs lacking the thumb residues fails to restore growth and fatty acid synthesis to a temperature-sensitive E. coli strain that lacks BCCP when grown at nonpermissive temperature. Alignment of BCCPs from various organisms shows that only two of the eight thumb residues are strictly conserved, and amino acid substitution of either residue results in proteins giving only weak growth of the temperature-sensitive E. coli strain. Therefore, the thumb structure is essential for the function of BCCP in the ACC reaction and provides a useful motif for distinguishing the biotinylated proteins of multisubunit ACCs from those of enzymes catalyzing other biotin-dependent reactions. An unexpected result was that expression of a mutant BCCP in which the biotinylated lysine residue was substituted with cysteine was able to partially restore growth and fatty acid synthesis to the temperature-sensitive E. coli strain. This complementation was shown to be specific to BCCPs having native structure (excepting the biotinylated lysine) and is interpreted in terms of dimerization of the BCCP biotinyl domain during the ACC reaction. PMID- 11495923 TI - Agonist-induced signaling, desensitization, and internalization of a phosphorylation-deficient AT1A angiotensin receptor. AB - An analysis of the functional role of a diacidic motif (Asp236-Asp237) in the third intracellular loop of the AT1A angiotensin II (Ang II) receptor (AT1-R) revealed that substitution of both amino acids with alanine (DD-AA) or asparagine (DD-NN) residues diminished Ang II-induced receptor phosphorylation in COS-7 cells. However, Ang II-stimulated inositol phosphate production, mitogen activated protein kinase, and AT1 receptor desensitization and internalization were not significantly impaired. Overexpression of dominant negative G protein coupled receptor kinase 2 (GRK2)K220M decreased agonist-induced receptor phosphorylation by approximately 40%, but did not further reduce the impaired phosphorylation of DD-AA and DD-NN receptors. Inhibition of protein kinase C by bisindolylmaleimide reduced the phosphorylation of both the wild-type and the DD mutant receptors by approximately 30%. The inhibitory effects of GRK2K220M expression and protein kinase C inhibition by bisindolylmaleimide on agonist induced phosphorylation were additive for the wild-type AT1-R, but not for the DD mutant receptor. Agonist-induced internalization of the wild-type and DD mutant receptors was similar and was unaltered by coexpression of GRK2K220M. These findings demonstrate that an acidic motif at position 236/237 in the third intracellular loop of the AT1-R is required for optimal Ang II-induced phosphorylation of its carboxyl-terminal tail by GRKs. Furthermore, the properties of the DD mutant receptor suggest that not only Ang II-induced signaling, but also receptor desensitization and internalization, are independent of agonist-induced GRK-mediated phosphorylation of the AT1 receptor. PMID- 11495924 TI - RGS9-G beta 5 substrate selectivity in photoreceptors. Opposing effects of constituent domains yield high affinity of RGS interaction with the G protein effector complex. AB - RGS proteins regulate the duration of G protein signaling by increasing the rate of GTP hydrolysis on G protein alpha subunits. The complex of RGS9 with type 5 G protein beta subunit (G beta 5) is abundant in photoreceptors, where it stimulates the GTPase activity of transducin. An important functional feature of RGS9-G beta 5 is its ability to activate transducin GTPase much more efficiently after transducin binds to its effector, cGMP phosphodiesterase. Here we show that different domains of RGS9-G beta 5 make opposite contributions toward this selectivity. G beta 5 bound to the G protein gamma subunit-like domain of RGS9 acts to reduce RGS9 affinity for transducin, whereas other structures restore this affinity specifically for the transducin-phosphodiesterase complex. We suggest that this mechanism may serve as a general principle conferring specificity of RGS protein action. PMID- 11495925 TI - Tumor necrosis factor-alpha-converting enzyme (ADAM17) mediates the cleavage and shedding of fractalkine (CX3CL1). AB - Fractalkine (CX3CL1) is an unusual member of the chemokine family that is synthesized with its chemokine domain at the end of a mucin-rich, transmembrane stalk. This membrane-bound localization allows fractalkine to function as an adhesion molecule for cells bearing its receptor, CX3CR1. In addition, fractalkine can be proteolytically released from the cell surface, generating a soluble molecule that functions as a chemoattractant similar to the other members of the chemokine family. In this study, we have examined the mechanisms that regulate the conversion between these two functionally distinct forms of fractalkine. We demonstrate that under normal conditions fractalkine is synthesized as an intracellular precursor that is rapidly transported to the cell surface where it becomes a target for metalloproteinase-dependent cleavage that causes the release of a fragment containing the majority of the fractalkine extracellular domain. We show that the cleavage of fractalkine can be markedly enhanced by stimulating cells with phorbol 12-myristate 13-acetate (PMA), and we identify tumor necrosis factor-alpha converting enzyme (TACE; ADAM17) as the protease responsible for this PMA-induced fractalkine release. In addition, we provide data showing that TACE-mediated fractalkine cleavage occurs at a site distinct from the dibasic juxtamembrane motif that had been suggested previously based on protein sequence homologies. The identification of TACE as a major protease responsible for the conversion of fractalkine from a membrane-bound adhesion molecule to a soluble chemoattractant will provide new information for understanding the physiological function of this chemokine. PMID- 11495926 TI - A polyketide synthase in glycopeptide biosynthesis: the biosynthesis of the non proteinogenic amino acid (S)-3,5-dihydroxyphenylglycine. AB - Balhimycin, a vancomycin-type antibiotic from Amycolatopsis mediterranei, contains the unusual amino acid (S)-3,5-dihydroxyphenylglycine (Dpg), with an acetate-derived carbon backbone. After sequence analysis of the biosynthetic gene cluster, one gene, dpgA, for a predicted polyketide synthase (PKS) was identified, sharing 20-30% identity with plant chalcone synthases. Inactivation of dpgA resulted in loss of balhimycin production, and restoration was achieved by supplementation with 3,5-dihydroxyphenylacetic acid, which is both a possible product of a PKS reaction and a likely precursor of Dpg. Enzyme assays with the protein expressed in Streptomyces lividans showed that this PKS uses only malonyl CoA as substrate to synthesize 3,5-dihydroxyphenylacetic acid. The PKS gene is organized in an operon-like structure with three downstream genes that are similar to enoyl-CoA-hydratase genes and a dehydrogenase gene. The heterologous co-expression of all four genes led to accumulation of 3,5 dihydroxyphenylglyoxylic acid. Therefore, we now propose a reaction sequence. The final step in the pathway to Dpg is a transamination. A predicted transaminase gene was inactivated, resulting in abolished antibiotic production and accumulation of 3,5-dihydroxyphenylglyoxylic acid. Interestingly, restoration was only possible by simultaneous supplementation with (S)-3,5-dihydroxyphenylglycine and (S)-4-hydroxyphenylglycine, indicating that the transaminase is essential for the formation of both amino acids. PMID- 11495927 TI - Peroxisome proliferator-activated receptor-alpha regulates lipid homeostasis, but is not associated with obesity: studies with congenic mouse lines. AB - Considerable controversy exists in determining the role of peroxisome proliferator-activated receptor-alpha (PPARalpha) in obesity. Two purebred congenic strains of PPARalpha-null mice were developed to study the role of this receptor in modulating lipid transport and storage. Weight gain and average body weight in wild-type and PPARalpha-null mice on either an Sv/129 or a C57BL/6N background were not markedly different between genotypes from 3 to 9 months of age. However, gonadal adipose stores were significantly greater in both strains of male and female PPARalpha-null mice. Hepatic accumulation of lipids was greater in both strains and sexes of PPARalpha-null mice compared with wild-type controls. Administration of the peroxisome proliferator WY-14643 caused hepatomegaly, alterations in mRNAs encoding proteins that regulate lipid metabolism, and reduced serum triglycerides in a PPARalpha-dependent mechanism. Constitutive differences in serum cholesterol and triglycerides in PPARalpha-null mice were found between genetic backgrounds. Results from this work establish that PPARalpha is a critical modulator of lipid homeostasis in two congenic mouse lines. This study demonstrates that disruption of the murine gene encoding PPARalpha results in significant alterations in constitutive serum, hepatic, and adipose tissue lipid metabolism. However, an overt, obese phenotype in either of the two congenic strains was not observed. In contrast to earlier published work, this study establishes that PPARalpha is not associated with obesity in mice. PMID- 11495928 TI - Molecular cloning and characterization of Atp6n1b: a novel fourth murine vacuolar H+-ATPase a-subunit gene. AB - The 116-kDa a-subunit of the vacuolar proton pump (H(+)-ATPase) exists as several isoforms encoded by different genes and with different patterns of tissue expression. Its function within the multisubunit pump complex has yet to be elucidated. To date, three isoforms have been identified in mouse (designated a1 a3). We now report the cloning and characterization of Atp6n1b, encoding a novel fourth murine isoform (a4). Murine a4 has 833 residues and shows 85% amino acid identity to the human kidney-specific ATP6N1B protein in which loss-of-function alterations cause autosomal recessive distal renal tubular acidosis. The human and murine genes have similar genomic organization; furthermore, Atp6n1b maps to a region of mouse chromosome 6 that is syntenic with the segment of human 7q33-34 containing ATP6N1B. Together these findings establish the two genes as orthologs. The mouse a4 protein is 61, 52, and 47% identical to a1, a2, and a3, respectively. Phylogenetic analysis confirms that among vertebrates there are four a-subunit families, with a4 most resembling a1. Northern blot analysis of Atp6n1b reveals a 3.7-kilobase a4 transcript in kidney but not other major organs, and a reverse transcription polymerase chain reaction in 12 mouse tissues detects expression in kidney alone. Immunofluorescence studies in murine kidney demonstrate high intensity a4 staining at the surface of intercalated cells, with additional expression in the proximal tubule (not previously reported in human kidney). Similar apical a4 immunostaining is also present in male genital tissue. Identification of this novel murine kidney-enriched 116-kDa a-subunit provides a molecular tool for investigation of the currently unknown role of this protein, which is essential for proper function of the apical renal vacuolar H(+)-ATPase in man. PMID- 11495929 TI - Evidence for an active process and a cochlear amplifier in nonmammals. AB - The last two decades have produced a great deal of evidence that in the mammalian organ of Corti outer hair cells undergo active shape changes that are part of a "cochlear amplifier" mechanism that increases sensitivity and frequency selectivity of the hearing epithelium. However, many signs of active processes have also been found in nonmammals, raising the question as to the ancestry and commonality of these mechanisms. Active movements would be advantageous in all kinds of sensory hair cells because they help signal detection at levels near those of thermal noise and also help to overcome fluid viscosity. Such active mechanisms therefore presumably arose in the earliest kinds of hair cells that were part of the lateral line system of fish. These cells were embedded in a firm epithelium and responded to relative motion between the hair bundle and the hair cell, making it highly likely that the first active motor mechanism was localized in the hair-cell bundle. In terrestrial nonmammals, there are many auditory phenomena that are best explained by the presence of a cochlear amplifier, indicating that in this respect the mammalian ear is not unique. The latest evidence supports siting the active process in nonmammals in the hair-cell bundle and in intimate association with the transduction process. PMID- 11495930 TI - Role of retinal slip in the prediction of target motion during smooth and saccadic pursuit. AB - Visual tracking of moving targets requires the combination of smooth pursuit eye movements with catch-up saccades. In primates, catch-up saccades usually take place only during pursuit initiation because pursuit gain is close to unity. This contrasts with the lower and more variable gain of smooth pursuit in cats, where smooth eye movements are intermingled with catch-up saccades during steady-state pursuit. In this paper, we studied in detail the role of retinal slip in the prediction of target motion during smooth and saccadic pursuit in the cat. We found that the typical pattern of pursuit in the cat was a combination of smooth eye movements with saccades. During smooth pursuit initiation, there was a correlation between peak eye acceleration and target velocity. During pursuit maintenance, eye velocity oscillated at approximately 3 Hz around a steady-state value. The average gain of smooth pursuit was approximately 0.5. Trained cats were able to continue pursuing in the absence of a visible target, suggesting a role of the prediction of future target motion in this species. The analysis of catch-up saccades showed that the smooth-pursuit motor command is added to the saccadic command during catch-up saccades and that both position error and retinal slip are taken into account in their programming. The influence of retinal slip on catch-up saccades showed that prediction about future target motion is used in the programming of catch-up saccades. Altogether, these results suggest that pursuit systems in primates and cats are qualitatively similar, with a lower average gain in the cat and that prediction affects both saccades and smooth eye movements during pursuit. PMID- 11495931 TI - Noise-enhanced heart rate and sympathetic nerve responses to oscillatory lower body negative pressure in humans. AB - By injecting noise into the carotid sinus baroreceptors, we previously showed that heart rate (HR) responses to weak oscillatory tilt were enhanced via a mechanism known as "stochastic resonance." It remains unclear, however, whether the same responses would be observed when using oscillatory lower body negative pressure (LBNP), which would unload the cardiopulmonary baroreceptors with physically negligible effects on the arterial system. Also, the vasomotor sympathetic activity directly controlling peripheral resistance against hypotensive stimuli was not observed. We therefore investigated the effects of weak (0 to approximately -10 mmHg) oscillatory (0.03 Hz) LBNP on HR and muscle sympathetic nerve activity (MSNA) while adding incremental noise to the carotid sinus baroreceptors via a pneumatic neck chamber. The signal-to-noise ratio of HR, cardiac interbeat interval, and total MSNA were all significantly improved by increasing noise intensity, while there was no significant change in the arterial blood pressure in synchronized with the oscillatory LBNP. We conclude that the stochastic resonance, affecting both HR and MSNA, results from the interaction of noise with the signal in the brain stem, where the neuronal inputs from the arterial and cardiopulmonary baroreceptors first come together in the nucleus tractus solitarius. Also, these results indicate that the noise could induce functional improvement in human blood pressure regulatory system in overcoming given hypotensive stimuli. PMID- 11495932 TI - Differential short-term changes in GABAergic or glycinergic synaptic efficacy on rat hypoglossal motoneurons. AB - Using whole cell patch-clamp recording from hypoglossal motoneurons of a neonatal rat brain slice preparation, we investigated short-term changes in synaptic transmission mediated by GABA or glycine. In 1.5 mM extracellular Ca(2+) [Ca(2+)](o), pharmacologically isolated GABAergic or glycinergic currents were elicited by electrical stimulation of the reticular formation. At low stimulation frequency, glycinergic currents were larger and faster than GABAergic ones. GABAergic currents were strongly facilitated by pulse trains at 5 or 10 Hz without apparent depression. This phenomenon persisted after pharmacological block of GABA(B) receptors. Glycinergic currents were comparatively much less enhanced than GABAergic currents. One possible mechanism to account for this difference is that GABAergic currents decayed so slowly that consecutive responses summated over an incrementing baseline. However, while synaptic summation appeared at > or =10-Hz stimulation, at 5 Hz strong facilitation developed with minimal summation of GABA-mediated currents. Glycinergic currents decayed so fast that summation was minimal. As [Ca(2+)](o) is known to shape short-term synaptic changes, we examined if varying [Ca(2+)](o) could differentially affect facilitation of GABA- or glycine-operated synapses. With 5 mM [Ca(2+)](o), the frequency of spontaneous GABAergic or glycinergic currents appeared much higher but GABAergic current facilitation was blocked (and replaced by depression), whereas glycinergic currents remained slightly facilitated. [Ca(2+)](o) manipulation thus brought about distinct processes responsible for facilitation of GABAergic or glycinergic transmission. Our data therefore demonstrate an unexpectedly robust, short-term increase in the efficiency of GABAergic synapses that can become at least as effective as glycinergic synapses. PMID- 11495933 TI - Somatosensory loss increases vestibulospinal sensitivity. AB - To determine whether subjects with somatosensory loss show a compensatory increase in sensitivity to vestibular stimulation, we compared the amplitude of postural lean in response to four different intensities of bipolar galvanic stimulation in subjects with diabetic peripheral neuropathy (PNP) and age-matched control subjects. To determine whether healthy and neuropathic subjects show similar increases in sensitivity to galvanic vestibular stimulation when standing on unstable surfaces, both groups were exposed to galvanic stimulation while standing on a compliant foam surface. In these experiments, a 3-s pulse of galvanic current was administered to subjects standing with eyes closed and their heads turned toward one shoulder (anodal current on the forward mastoid). Anterior body tilt, as measured by center of foot pressure (CoP), increased proportionately with increasing galvanic vestibular stimulation intensity for all subjects. Subjects with peripheral neuropathy showed larger forward CoP displacement in response to galvanic stimulation than control subjects. The largest differences between neuropathy and control subjects were at the highest galvanic intensities, indicating an increased sensitivity to vestibular stimulation. Neuropathy subjects showed a larger increase in sensitivity to vestibular stimulation when standing on compliant foam than control subjects. The effect of galvanic stimulation was larger on the movement of the trunk segment in space than on the body's center of mass (CoM) angle, suggesting that the vestibular system acts to control trunk orientation rather than to control whole body posture. This study provides evidence for an increase in the sensitivity of the postural control system to vestibular stimulation when somatosensory information from the surface is disrupted either by peripheral neuropathy or by standing on an unstable surface. Simulations from a simple model of postural orientation incorporating feedback from the vestibular and somatosensory systems suggest that the increase in body lean in response to galvanic current in subjects with neuropathy could be reproduced only if central vestibular gain was increased when peripheral somatosensory gain was decreased. The larger effects of galvanic vestibular stimulation on the trunk than on the body's CoM suggest that the vestibular system may act to control postural orientation via control of the trunk in space. PMID- 11495934 TI - Multiple effects of dopamine on layer V pyramidal cell excitability in rat prefrontal cortex. AB - The mechanisms underlying the inhibitory effects of dopamine (DA) on layer V pyramidal neuron excitability in the prelimbic region of the rat medial prefrontal cortex were investigated. Under control conditions, DA depressed both action potential generation (driven by somatic current injection) and input resistance (R(N)). The presence of GABA(A) receptor antagonists blocked DA induced depression of action potential generation and revealed a delayed increase in excitability that persisted for the duration of experimental recording, up to 20 min following the washout of DA. In contrast to spike generation, disinhibition did not affect the transient depression of R(N) produced by DA, suggesting independent actions of DA on spike generation and R(N). Consistent with the hypothesis that DA acts to decrease pyramidal cell output via a GABAergic mechanism, DA increased the frequency of spontaneous inhibitory postsynaptic currents in both the absence and presence of TTX. Furthermore focal application of GABA to a perisomatic region mimicked the inhibitory effect of DA on spike production without affecting R(N). Focal application of bicuculline to the same location reversed the inhibitory effect of bath-applied DA on spike generation, while again having no effect on R(N). The depression of R(N) by DA was both occluded and mimicked by the Na(+) channel blocker TTX, suggesting the involvement of a Na(+) conductance in reducing pyramidal cell R(N) during the acute presence of DA. Together these data demonstrate that the acute presence of DA decreases pyramidal neuron excitability by two independent mechanisms. At the same time DA triggers a delayed and longer-lasting increase in excitability that is partially masked by synaptic inhibition. PMID- 11495935 TI - Paradoxical reduction of synaptic inhibition by vigabatrin. AB - GABAergic inhibition, a primary target for pharmacological modulation of excitability in the CNS, can be altered by multiple mechanisms including alteration of GABA metabolism. Gamma-vinyl GABA (vigabatrin, GVG) is an irreversible inhibitor of the GABA catabolic enzyme GABA transaminase, thus its anticonvulsant properties are thought to result from an elevation of brain GABA levels. We examined the effects of GVG on GABAergic synaptic transmission in hippocampal slices. GVG unexpectedly reduced miniature and evoked inhibitory postsynaptic currents (IPSCs) in dentate granule cells. The reduction in synaptic events was accompanied by an increase in tonic GABA(A) receptor-mediated current. These effects developed slowly and persisted following wash out of GVG. The GVG pretreatment reduced sucrose-evoked GABA release as well as postsynaptic sensitivity to exogenous GABA, indicating that both pre- and postsynaptic mechanisms contributed to the reduction in synaptic currents. These results suggest that tonic rather than phasic increases in GABA underlie the anticonvulsant properties of GVG, and that mechanisms that elevate brain neurotransmitter levels do not necessarily correlate with enhanced synaptic release. PMID- 11495936 TI - Two virtual fingers in the control of the tripod grasp. AB - To investigate the organization of multi-fingered grasping, we asked subjects to grasp an object using three digits: the thumb, the index finger, and the middle or ring finger. The object had three coarse flat contact surfaces, whose locations and orientations were varied systematically. Subjects were asked to grasp and lift the object and then to hold it statically. We analyzed the grasp forces in the horizontal plane that were recorded during the static hold period. Static equilibrium requires that the forces exerted by the three digits intersect at a common point, the force focus. The directions of the forces exerted by the two fingers opposing the thumb depended on the orientation of the contact surfaces of both fingers but not on the orientation of the contact surface of the thumb. The direction of the thumb's force did not depend on the orientation of the contact surfaces of the two fingers and depended only weakly on the orientation of the thumb's contact surface. In general, the thumb's force was directed to a point midway between the two fingers. The results are consistent with a hierarchical model of the control of a tripod grasp. At the first level, an opposition space is created between the thumb and a virtual finger located approximately midway between the two actual fingers. The directions of the forces exerted by the two fingers are constrained to be mirror symmetric about the opposition axis. The actual directions of finger force are elaborated at the next level on the basis of stability considerations. PMID- 11495937 TI - Recruitment of cat motoneurons in the absence of homonymous afferent feedback. AB - This study provides the first test in vivo of the hypothesis that group Ia muscle stretch afferents aid in preventing reversals in the orderly recruitment of motoneurons. This hypothesis was tested by studying recruitment of motoneurons deprived of homonymous afferent input. Recruitment order was measured in decerebrate, paralyzed cats from dual intra-axonal records obtained simultaneously from pairs of medial gastrocnemius (MG) motoneurons. Pairs of MG motor axons were recruited in eight separate trials of the reflex discharge evoked by stimulation of the caudal cutaneous sural (CCS) nerve. Some reports suggest that reflex recruitment by this cutaneous input should bias recruitment against order by the size principle in which the axon with the slower conduction velocity (CV) in a pair is recruited to fire before the faster CV axon. Recruitment was studied in three groups of cats: ones with the MG nerve intact and untreated (UNTREATED); ones with the MG nerve cut (CUT); and ones with the MG nerve cut and bathed at its proximal end in lidocaine solution (CUT+). The failure of electrical stimulation to initiate a dorsal root volley and the absence of action potentials in MG afferents demonstrated the effective elimination of afferent feedback in the CUT+ group. Recruitment order by the size principle predominated and was not statistically distinguishable among the three groups. The percentage of pairs recruited in reverse order of the size principle was actually smaller in the CUT+ group (6%) than in CUT (15%) or UNTREATED (19%) groups. Thus homonymous afferent feedback is not necessary to prevent recruitment reversal. However, removing homonymous afferent input did result in the expression of inconsistency in order, i.e., switches in recruitment sequence from one trial to the next, for more axon pairs in the CUT+ group (33%) than for the other groups combined (13%). Increased inconsistency in the absence of increased reversal of recruitment order was approximated in computer simulations by increasing time-varying fluctuations in synaptic drive to motoneurons and could not be reproduced simply by deleting synaptic current from group Ia homonymous afferents, regardless of how that current was distributed to the motoneurons. These findings reject the hypothesis that synaptic input from homonymous group Ia afferents is necessary to prevent recruitment reversals, and they are consistent with the assertion that recruitment order is established predominantly by properties intrinsic to motoneurons. PMID- 11495938 TI - Contribution of Na(v)1.8 sodium channels to action potential electrogenesis in DRG neurons. AB - C-type dorsal root ganglion (DRG) neurons can generate tetrodotoxin-resistant (TTX-R) sodium-dependent action potentials. However, multiple sodium channels are expressed in these neurons, and the molecular identity of the TTX-R sodium channels that contribute to action potential production in these neurons has not been established. In this study, we used current-clamp recordings to compare action potential electrogenesis in Na(v)1.8 (+/+) and (-/-) small DRG neurons maintained for 2-8 h in vitro to examine the role of sodium channel Na(v)1.8 (alpha-SNS) in action potential electrogenesis. Although there was no significant difference in resting membrane potential, input resistance, current threshold, or voltage threshold in Na(v)1.8 (+/+) and (-/-) DRG neurons, there were significant differences in action potential electrogenesis. Most Na(v)1.8 (+/+) neurons generate all-or-none action potentials, whereas most of Na(v)1.8 (-/-) neurons produce smaller graded responses. The peak of the response was significantly reduced in Na(v)1.8 (-/-) neurons [31.5 +/- 2.2 (SE) mV] compared with Na(v)1.8 (+/+) neurons (55.0 +/- 4.3 mV). The maximum rise slope was 84.7 +/- 11.2 mV/ms in Na(v)1.8 (+/+) neurons, significantly faster than in Na(v)1.8 (-/-) neurons where it was 47.2 +/- 1.3 mV/ms. Calculations based on the action potential overshoot in Na(v)1.8 (+/+) and (-/-) neurons, following blockade of Ca(2+) currents, indicate that Na(v)1.8 contributes a substantial fraction (80-90%) of the inward membrane current that flows during the rising phase of the action potential. We found that fast TTX-sensitive Na(+) channels can produce all-or none action potentials in some Na(v)1.8 (-/-) neurons but, presumably as a result of steady-state inactivation of these channels, electrogenesis in Na(v)1.8 (-/-) neurons is more sensitive to membrane depolarization than in Na(v)1.8 (+/+) neurons, and, in the absence of Na(v)1.8, is attenuated with even modest depolarization. These observations indicate that Na(v)1.8 contributes substantially to action potential electrogenesis in C-type DRG neurons. PMID- 11495939 TI - Nerve growth factor regulates sodium but not potassium channel currents in sympathetic B neurons of adult bullfrogs. AB - The TTX-sensitive and -resistant components of the voltage-gated Na(+) current (TTX-s I(Na) and TTX-r I(Na)) are increased within 2 wk of cutting the axons of B cells in bullfrog paravertebral sympathetic ganglia (BFSG). Axotomy also increases the noninactivating, voltage-activated K(+) current (M current I(M)), whereas delayed rectifier K(+) current (I(K)) is reduced. We found that similar effects were produced when BFSG B cells were dissociated from adult bullfrogs and maintained in a defined-medium, neuron-enriched, low-density, serum-free culture. Thus the density of TTX-s I(Na), TTX-r I(Na), and I(M) were transiently increased, whereas I(K) density was decreased. Reduction in voltage-sensitive, Ca(2+)-dependent K(+) current (I(C)) was attributed to previously documented decreases in Ca(2+) channel current (I(Ca)). To test whether axotomy- or culture induced changes in ion channel function reflect loss of retrograde influence of nerve growth factor (NGF), we examined the effect of murine beta-NGF on TTX-s I(Na), TTX-r I(Na), I(K), and I(M). Culture of neurons for 15 days in the presence of NGF (200 ng/ml), more than doubled total I(Na) density but did not enhance neurite outgrowth. The TTX-r I(Na) density was increased about threefold and the TTX-s I(Na) density increased 2.4-fold. NGF did not affect the activation or inactivation kinetics of the total Na(+) conductance. Effects of NGF were blocked by the transcription inhibitors, cordycepin (20 microM) and actinomycin D (0.01 microg/ml). I(K) and I(M) were unaffected by NGF, and although I(C) was enhanced, this likely reflected the known effect of NGF on I(Ca) in BFSG neurons. Na(+) channel synthesis and/or expression in adult sympathetic neurons is therefore subject to selective regulation by NGF. Despite this, the increase in I(Na) and I(M) as well as the decrease in I(K) seen in BFSG neurons in culture or after axotomy cannot readily be explained in terms of alterations in the availability of target-derived NGF. PMID- 11495940 TI - Gaze-stabilizing deficits and latent nystagmus in monkeys with brief, early-onset visual deprivation: eye movement recordings. AB - The normal development and the capacity to calibrate gaze-stabilizing systems may depend on normal vision during infancy. At the end of 1 yr of dark rearing, cats have gaze-stabilizing deficits similar to that of the newborn human infant including decreased monocular optokinetic nystagmus (OKN) in the nasal to temporal (N-T) direction and decreased velocity storage in the vestibuloocular reflex (VOR). The purpose of this study is to determine to what extent restricted vision during the first 2 mo of life in monkeys affects the development of gaze stabilizing systems. The eyelids of both eyes were sutured closed in three rhesus monkeys (Macaca mulatta) at birth. Eyelids were opened at 25 days in one monkey and 40 and 55 days in the other two animals. Eye movements were recorded from each eye using scleral search coils. The VOR, OKN, and fixation were examined at 6 and 12 mo of age. We also examined ocular alignment, refraction, and visual acuity in these animals. At 1 yr of age, visual acuity ranged from 0.3 to 0.6 LogMAR (20/40-20/80). All animals showed a defect in monocular OKN in the N-T direction. The velocity-storage component of OKN (i.e., OKAN) was the most impaired. All animals had a mild reduction in VOR gain but had a normal time constant. The animals deprived for 40 and 55 days had a persistent strabismus. All animals showed a nystagmus similar to latent nystagmus (LN) in human subjects. The amount of LN and OKN defect correlated positively with the duration of deprivation. In addition, the animal deprived for 55 days demonstrated a pattern of nystagmus similar to congenital nystagmus in human subjects. We found that restricted visual input during the first 2 mo of life impairs certain gaze stabilizing systems and causes LN in primates. PMID- 11495941 TI - Gaze-stabilizing deficits and latent nystagmus in monkeys with early-onset visual deprivation: role of the pretectal not. AB - We studied the role of the pretectal nucleus of the optic tract (NOT) in the development of monocular optokinetic nystagmus (OKN) asymmetries and latent nystagmus (LN) in two monkeys reared with binocular deprivation (BD) caused by binocular eyelid suture for either the first 25 or 55 days of life. Single-unit recordings were performed in the right and left NOT of both monkeys at 2-3 yr of age and compared with similar unit recordings in normally reared monkeys. We also examined ocular motor behavior during electrical stimulation of the NOT and during pharmacological inactivation and activation using GABA(A) agonists and antagonists. In BD animals a large proportion of NOT units was dominated by the contralateral eye, in striking contrast to normal animals where 100% of NOT units were sensitive to stimuli delivered to either eye. In the 55-day BD animal no binocularly sensitive neurons were found, while in the 25-day BD animal 60% of NOT units retained at least some binocular sensitivity. Differences in direction sensitivity were also observed in BD animals. We found that 56% of units in the 55-day BD monkey and 10% of units in the 25-day BD monkey responded preferentially to contraversive visual motion. In contrast, only 5% of the NOT units encountered in normally reared monkeys respond preferentially during contraversive visual motion, the rest were most sensitive to ipsiversive visual motion. NOT neurons of BD monkeys showed a wide range of speed sensitivities similar to that of normal monkeys. Unilateral electrical stimulation of the NOT in BD animals induced a conjugate nystagmus with slow phases directed toward the side of stimulation. When we blocked the activity of NOT units with muscimol, a potent GABA(A) agonist, LN was abolished. In contrast, LN was increased when spontaneous activity of the NOT was enhanced with bicuculline, a GABA(A) antagonist. Our results indicate that the NOT in BD monkeys plays an important role in the OKN deficits and LN generation during monocular viewing. We hypothesize that the large proportion of units dominated by the contralateral eye contribute to the development of monocular OKN asymmetries and LN. PMID- 11495942 TI - Dependence of saccade-related activity in the primate superior colliculus on visual target presence. AB - Neurons in the intermediate layers of the superior colliculus respond to visual targets and/or discharge immediately before and during saccades. These visual and motor responses have generally been considered independent, with the visual response dependent on the nature of the stimulus, and the saccade-related activity related to the attributes of the saccade, but not to how the saccade was elicited. In these experiments we asked whether saccade-related discharge in the superior colliculus depended on whether the saccade was directed to a visual target. We recorded extracellular activity of neurons in the intermediate layers of the superior colliculus of three rhesus monkeys during saccades in tasks in which we varied the presence or absence of a visual target and the temporal delays between the appearance and disappearance of a target and saccade initiation. Across our sample of neurons (n = 64), discharge was highest when a saccade was made to a still-present visual target, regardless of whether the target had recently appeared or had been present for several hundred milliseconds. Discharge was intermediate when the target had recently disappeared and lowest when the target had never appeared during that trial. These results are consistent with the hypothesis that saccade-related discharge decreases as the time between the target disappearance and saccade initiation increases. Saccade velocity was also higher for saccades to visual targets, and correlated on a trial-by-trial basis with perisaccadic discharge for many neurons. However, discharge of many neurons was dependent on task but independent of saccade velocity, and across our sample of neurons, saccade velocity was higher for saccades made immediately after target appearance than would be predicted by discharge level. A tighter relationship was found between saccade precision and perisaccadic discharge. These findings suggest that just as the purpose of the saccadic system in primates is to drive the fovea to a visual target, presaccadic motor activity in the superior colliculus is most intense when such a target is actually present. This enhanced activity may, itself, contribute to the enhanced performance of the saccade system when the saccade is made to a real visual target. PMID- 11495943 TI - Active locomotion increases peak firing rates of anterodorsal thalamic head direction cells. AB - Head direction (HD) cells discharge selectively in macaques, rats, and mice when they orient their head in a specific ("preferred") direction. Preferred directions are influenced by visual cues as well as idiothetic self-motion cues derived from vestibular, proprioceptive, motor efferent copy, and command signals. To distinguish the relative importance of active locomotor signals, we compared HD cell response properties in 49 anterodorsal thalamic HD cells of six male Long-Evans rats during active displacements in a foraging task as well as during passive rotations. Since thalamic HD cells typically stop firing if the animals are tightly restrained, the rats were trained to remain immobile while drinking water distributed at intervals from a small reservoir at the center of a rotatable platform. The platform was rotated in a clockwise/counterclockwise oscillation to record directional responses in the stationary animals while the surrounding environmental cues remained stable. The peak rate of directional firing decreased by 27% on average during passive rotations (r(2) = 0.73, P < 0.001). Individual cells recorded in sequential sessions (n = 8) reliably showed comparable reductions in peak firing, but simultaneously recorded cells did not necessarily produce identical responses. All of the HD cells maintained the same preferred directions during passive rotations. These results are consistent with the hypothesis that the level of locomotor activity provides a state-dependent modulation of the response magnitude of AD HD cells. This could result from diffusely projecting neuromodulatory systems associated with motor state. PMID- 11495944 TI - Resonance of spike discharge modulation in neurons of the guinea pig medial vestibular nucleus. AB - The modulation of action potential discharge rates is an important aspect of neuronal information processing. In these experiments, we have attempted to determine how effectively spike discharge modulation reflects changes in the membrane potential in central vestibular neurons. We have measured how their spike discharge rate was modulated by various current inputs to obtain neuronal transfer functions. Differences in the modulation of spiking rates were observed between neurons with a single, prominent after hyperpolarization (AHP, type A neurons) and cells with more complex AHPs (type B neurons). The spike discharge modulation amplitudes increased with the frequency of the current stimulus, which was quantitatively described by a neuronal model that showed a resonance peak >10 Hz. Modeling of the resonance peak required two putative potassium conductances whose properties had to be markedly dependent on the level of the membrane potential. At low frequencies (< or =0.4 Hz), the gain or magnitude functions of type A and B discharge rates were similar relative to the current input. However, resting input resistances obtained from the ratio of the membrane potential and current were lower in type B compared with type A cells, presumably due to a higher level of active potassium conductances at rest. The lower input resistance of type B neurons was compensated by a twofold greater sensitivity of their firing rate to changes in membrane potential, which suggests that synaptic inputs on their dendritic processes would be more efficacious. This increased sensitivity is also reflected in a greater ability of type B neurons to synchronize with low-amplitude sinusoidal current inputs, and in addition, their responses to steep slope ramp stimulation are enhanced over the more linear behavior of type A neurons. This behavior suggests that the type B MVNn are moderately tuned active filters that promote high-frequency responses and that type A neurons are like low-pass filters that are well suited for the resting tonic activity of the vestibular system. However, the more sensitive and phasic type B neurons contribute to both low- and high-frequency control as well as signal detection and would amplify the contribution of both irregular and regular primary afferents at high frequencies. PMID- 11495945 TI - Pathway-specific targeting of GABA(A) receptor subtypes to somatic and dendritic synapses in the central amygdala. AB - Neurons in the central amygdala express two distinct types of ionotropic GABA receptor. One is the classical GABA(A) receptor that is blocked by low concentrations of bicuculline and positively modulated by benzodiazepines. The other is a novel type of ionotropic GABA receptor that is less sensitive to bicuculline but blocked by the GABA(C) receptor antagonist (1,2,5,6 tetrohydropyridine-4-yl) methylphosphinic acid (TPMPA) and by benzodiazepines. In this study, we examine the distribution of these two receptor types. Recordings of GABAergic miniature inhibitory postsynaptic currents (mIPSCs) showed a wide variation in amplitude. Most events had amplitudes of < 50 pA, but a small minority had amplitudes >100 pA. Large-amplitude events also had rise times faster than small-amplitude events. Large-amplitude events were fully blocked by 10 microM bicuculline but unaffected by TPMPA. Small amplitude events were partially blocked by both bicuculline and TPMPA. Focal application of hypertonic sucrose to the soma evoked large-amplitude mIPSCs, whereas focal dendritic application of sucrose evoked small-amplitude mIPSCs. Thus inhibitory synapses on the dendrites of neurons in the central amygdala express both types of GABA receptor, but somatic synapses expressed purely GABA(A) receptors. Minimal stimulation revealed that inhibitory inputs arising from the laterally located intercalated cells innervate dendritic synapses, whereas inhibitory inputs of medial origin innervated somatic inhibitory synapses. These results show that different types of ionotropic GABA receptors are targeted to spatially and functionally distinct synapses. Thus benzodiazepines will have different modulatory effects on different inhibitory pathways in the central amygdala. PMID- 11495946 TI - Analysis of the frequency response of the saccadic circuit: system behavior. AB - To more thoroughly describe the system dynamics for the saccadic circuit in monkeys, we have determined the frequency response by applying a frequency modulated train of microstimulation pulses in the superior colliculus. The resulting eye movements reflect the transfer function of the saccadic circuit. Below input modulations of 5 cycles/s, the saccadic circuit increasingly oscillates with multiple high-frequency, low-amplitude movements reminiscent of the "staircase saccades" evoked during the sustained step response. Between 5 and 20 cycles/s, the circuit entrains well to the input, exhibiting one saccadic response to each sinusoidal input. Within this range there are systematic frequency-dependent changes in movement amplitudes, including super-normal saccades at some input frequencies. Above 20 cycles/s, the saccadic circuit increasingly exhibits periodic failures at rates of 1:2 or higher. In addition, the circuit exhibits predictable amplitude-modulated interference patterns in response to a combined step and frequency-modulated input. These experimental results provide insight into several biological mechanisms and serve as benchmark tests of viable models of the saccadic system. The data are consistent with negative feedback models of the saccadic system that operate as a displacement controller and inconsistent with theories that put the superior colliculus within the lowest-order, local feedback loop. The data support theories that the circuit feedback operates with dynamics that simulate a "leaky integrator." In addition, the results demonstrate how the temporal output of the superior colliculus interacts with recurrent inhibition to influence the eye movement dynamics. PMID- 11495947 TI - Experimental and computational analysis of monkey smooth pursuit eye movements. AB - Smooth pursuit eye movements are guided by visual feedback and are surprisingly accurate despite the time delay between visual input and motor output. Previous models have reproduced the accuracy of pursuit either by using elaborate visual signals or by adding sources of motor feedback. Our goal was to constrain what types of signals drive pursuit by obtaining data that would discriminate between these two modeling approaches, represented by the "image motion model" and the "tachometer feedback" model. Our first set of experiments probed the visual properties of pursuit with brief square-pulse and sine-wave perturbations of target velocity. Responses to pulse perturbations increased almost linearly with pulse amplitude, while responses to sine wave perturbations showed strong saturation with increasing stimulus amplitude. The response to sine wave perturbations was strongly dependent on the baseline image velocity at the time of the perturbation. Responses were much smaller if baseline image velocity was naturally large, or was artificially increased by superimposing sine waves on pulse perturbations. The image motion model, but not the tachometer feedback model, could reproduce these features of pursuit. We used a revision of the image motion model that was, like the original, sensitive to both image velocity and image acceleration. Due to a saturating nonlinearity, the sensitivity to image acceleration declined with increasing image velocity. Inclusion of this nonlinearity was motivated by our experimental results, was critical in accounting for the responses to perturbations, and provided an explanation for the unexpected stability of pursuit in the presence of perturbations near the resonant frequency. As an emergent property, the revised image motion model was able to reproduce the frequency and damping of oscillations recorded during artificial feedback delays. Our second set of experiments replicated prior recordings of pursuit responses to multiple-cycle sine wave perturbations, presented over a range of frequencies. The image motion model was able to reproduce the responses to sine wave perturbations across all frequencies, while the tachometer feedback model failed at high frequencies. These failures resulted from the absence of image acceleration signals in the tachometer model. We conclude that visual signals related to image acceleration are important in driving pursuit eye movements and that the nonlinearity of these signals provides stability. Smooth pursuit thus illustrates that a plausible neural strategy for combating natural delays in sensory feedback is to employ information about the derivative of the sensory input. PMID- 11495948 TI - Impaired inhibitory G-protein function contributes to increased calcium currents in rats with diabetic neuropathy. AB - There is a growing body of evidence that sensory neuropathy in diabetes is associated with abnormal calcium signaling in dorsal root ganglion (DRG) neurons. Enhanced influx of calcium via multiple high-threshold calcium currents is present in sensory neurons of several models of diabetes mellitus, including the spontaneously diabetic BioBred/Worchester (BB/W) rat and the chemical streptozotocin (STZ)-induced rat. We believe that abnormal calcium signaling in diabetes has pathologic significance as elevation of calcium influx and cytosolic calcium release has been implicated in other neurodegenerative conditions characterized by neuronal dysfunction and death. Using electrophysiologic and pharmacologic techniques, the present study provides evidence that significant impairment of G-protein-coupled modulation of calcium channel function may underlie the enhanced calcium entry in diabetes. N- and P-type voltage-activated, high-threshold calcium channels in DRGs are coupled to mu opiate receptors via inhibitory G(o)-type G proteins. The responsiveness of this receptor coupled model was tested in dorsal root ganglion (DRG) neurons from spontaneously diabetic BB/W rats, and streptozotocin-induced (STZ) diabetic rats. Intracellular dialysis with GTPgammaS decreased calcium current amplitude in diabetic BB/W DRG neurons compared with those of age-matched, nondiabetic controls, suggesting that inhibitory G-protein activity was diminished in diabetes, resulting in larger calcium currents. Facilitation of calcium current density (I(DCa)) by large amplitude depolarizing prepulses (proposed to transiently inactivate G proteins), was significantly less effective in neurons from BB/W and STZ-induced diabetic DRGs. Facilitation was enhanced by intracellular dialysis with GTPgammaS, decreased by pertussis toxin, and abolished by GDPbetaS within 5 min. Direct measurement of GTPase activity using opiate-mediated GTPgamma[(35)S] binding, confirmed that G-protein activity was significantly diminished in STZ-induced diabetic neurons compared with age-matched nondiabetic controls. Diabetes did not alter the level of expression of mu opiate receptors and G-protein alpha subunits. These studies indicate that impaired regulation of calcium channels by G proteins is an important mechanism contributing to enhanced calcium influx in diabetes. PMID- 11495949 TI - Ionic mechanisms underlying burst firing of layer III sensorimotor cortical neurons of the cat: an in vitro slice study. AB - We examined the ionic mechanisms underlying burst firing in layer III neurons from cat sensorimotor cortex by intracellular recording in a brain slice. Regular spiking was observed in 77.4% of 137 neurons in response to constant intracellular current pulses of 0.5- to 1-s duration. The rest of the neurons showed burst firing. An initial burst followed by regular-spike firing was seen in 71.0% of 31 bursting neurons. The rest of the bursting neurons (n = 9) exhibited repetitive bursting. In the bursting neurons, spikes comprising the burst were triggered from the afterdepolarization (ADP) of the first spike of the burst. We examined the ionic mechanisms underlying the ADP by applying channel blocking agents. The ADP was enhanced (rather than blocked) by Ca2+ channel blockade. This enhancement of the ADP by Ca2+ channel blockade was apparent even after blockade of the afterhyperpolarization by apamin or intracellular Ca2+ chelation by EGTA. The firing rate of the regular-spiking cells was increased by apamin, intracellular EGTA or Ca2+ channel blockers. In 17.9% of the neurons examined (n = 56), these agents switched the regular-spiking pattern into a bursting one. Burst firing could not be changed to regular spiking by these agents. Four neurons that responded with a single initial burst in control solution responded with repetitive bursting after application of these agents. We conclude that the main function of Ca2+ influx in layer III neurons is to activate Ca2+-dependent K+ conductance, which prevents or limits burst firing. At a time when spike amplitude was unchanged, the ADP was blocked and the burst firing changed to regular spiking by extracellularly applied tetrodotoxin (TTX) or intracellularly applied N-(2,6-dimethylphenylcarbamoylmethyl) triethyl ammonium bromide (QX314). We concluded that a TTX- and QX314-sensitive Na+ current underlies the ADP and therefore contributes to the burst firing of layer III neurons from the cat cortex. PMID- 11495950 TI - Sex-related differences in human pain and rat afferent discharge evoked by injection of glutamate into the masseter muscle. AB - Animal studies have suggested that tissue injury-related increased levels of glutamate may be involved in peripheral nociceptive mechanisms in deep craniofacial tissues. Indeed, injection of glutamate (0.1-1 M, 10 microl) into the temporomandibular region evokes reflex jaw muscle responses through activation of peripheral excitatory amino acid receptors. It has recently been found that this glutamate-evoked reflex muscle activity is significantly greater in female than male rats. However, it is not known whether peripheral administration of glutamate, in the same concentrations that evoke jaw muscle activity in rats, causes pain in humans or activates deep craniofacial nociceptive afferents. Therefore we examined whether injection of glutamate into the masseter muscle induces pain in male and female volunteers and, since masseter afferent recordings were not feasible in humans, whether glutamate excites putative nociceptive afferents supplying the masseter muscle of male and female rats. Injection of glutamate (0.5 M or 1.0 M, 0.2 ml) into the masseter muscle of both men and women caused significantly higher levels of peak pain, duration of pain, and overall pain than injection of isotonic saline (0.2 ml). In addition, glutamate-evoked peak and overall muscle pain in women was significantly greater than in men. In rats of both sexes, glutamate (10 microl, 0.5 M) evoked activity in a subpopulation of masseter muscle afferents (n = 36) that projected to the subnucleus caudalis, an important relay of noxious input from the craniofacial region. The largest responses to glutamate were recorded in muscle afferents with the slowest conduction velocities (2.5-5 m/s). Further, glutamate-evoked masseter muscle afferent activity was significantly greater in female than in male rats. These results indicate that glutamate injection into the masseter muscle evokes pain responses that are greater in women than men and that one possible mechanism for this difference may be a greater sensitivity to glutamate of masseter muscle afferents in females. These sex-related differences in acute experimental masseter muscle pain are particularly interesting given the higher prevalence of many chronic muscle pain conditions in women. PMID- 11495951 TI - Activation and reconfiguration of fictive feeding by the octopamine-containing modulatory OC interneurons in the snail Lymnaea. AB - We describe the role of the octopamine-containing OC interneurons in the buccal feeding system of Lymnaea stagnalis. OC neurons are swallowing phase interneurons receiving inhibitory inputs in the N1 and N2 phases, and excitatory inputs in the N3 phase of fictive feeding. Although the OC neurons do not always fire during feeding, the feeding rate is significantly (P < 0.001) higher when both SO and OC fire in each cycle than when only the SO fires. In 28% of silent preparations, a single stimulation of an OC interneuron evokes the feeding pattern. Repetitive stimulation of the OC interneuron increases the proportion of responsive preparations to 41%. The OC interneuron not only changes both the feeding rate and reconfigures the pattern. Depolarization of the OC interneurons increases the feeding rate and removes the B3 motor neuron from the firing sequence. Hyperpolarization slows it down (increasing the duration of N1 and N3 phases) and recruits the B3 motor neuron. OC interneurons form synaptic connections onto buccal motor neurons and interneurons but not onto the cerebral (cerebral giant cell) modulatory neurons. OC interneurons are electrically coupled to all N3 phase (B4, B4Cl, B8) feeding motor neurons. They form symmetrical connections with the N3p interneurons having dual electrical (excitatory) and chemical (inhibitory) components. OC interneurons evoke biphasic synaptic inputs on the protraction phase interneurons (SO, N1L, N1M), with a short inhibition followed by a longer lasting depolarization. N2d interneurons are hyperpolarized, while N2v interneurons are slowly depolarized and often fire a burst after OC stimulation. Most motor neurons also receive synaptic responses from the OC interneurons. Although OC and N3p interneurons are both swallowing phase interneurons, their synaptic contacts onto follower neurons are usually different (e.g., the B3 motor neurons are inhibited by OC, but excited by N3p interneurons). Repetitive stimulation of OC interneuron facilitates the excitatory component of the biphasic responses evoked on the SO, N1L, and N1M interneurons, but neither the N2 nor the N3 phase interneurons display a similar longer-lasting excitatory effect. OC interneurons are inhibited by all the buccal feeding interneurons, but excited by the serotonergic modulatory CGC neurons. We conclude that OC interneurons are a new kind of swallowing phase interneurons. Their connections with the buccal feeding interneurons can account for their modulatory effects on the feeding rhythm. As they contain octopamine, this is the first example in Lymnaea that monoaminergic modulation and reconfiguration are provided by an intrinsic member of the buccal feeding network. PMID- 11495952 TI - Mapping Pavlovian conditioning effects on the brain: blocking, contiguity, and excitatory effects. AB - Pavlovian conditioning effects on the brain were investigated by mapping rat brain activity with fluorodeoxyglucose (FDG) autoradiography. The goal was to map the effects of the same tone after blocking or eliciting a conditioned emotional response (CER). In the tone-blocked group, previous learning about a light blocked a CER to the tone. In the tone-excitor group, the same pairings of tone with shock US resulted in a CER to the tone in the absence of previous learning about the light. A third group showed no CER after pseudorandom presentations of these stimuli. Brain systems involved in the various associative effects of Pavlovian conditioning were identified, and their functional significance was interpreted in light of previous FDG studies. Three conditioning effects were mapped: 1) blocking effects: FDG uptake was lower in medial prefrontal cortex and higher in spinal trigeminal and cuneate nuclei in the tone-blocked group relative to the tone-excitor group. 2) Contiguity effects: relative to pseudorandom controls, similar FDG uptake increases in the tone-blocked and -excitor groups were found in auditory regions (inferior colliculus and cortex), hippocampus (CA1), cerebellum, caudate putamen, and solitary nucleus. Contiguity effects may be due to tone-shock pairings common to the tone-blocked and -excitor groups rather than their different CER. And 3) excitatory effects: FDG uptake increases limited to the tone-excitor group occurred in a circuit linked to the CER, including insular and anterior cingulate cortex, vertical diagonal band nucleus, anterior hypothalamus, and caudoventral caudate putamen. This study provided the first large-scale map of brain regions underlying the Kamin blocking effect on conditioning. In particular, the results suggest that suppression of prefrontal activity and activation of unconditioned stimulus pathways are important neural substrates of the Kamin blocking effect. PMID- 11495953 TI - Functional analyses of the leech swim oscillator. AB - The oscillations that underlie swimming movements in the leech arise from a series of identified concatenated circuits within the ventral nerve cord. In the intact nerve cord, ascending and descending intersegmental interactions via axons within the lateral connectives function both to generate robust oscillations throughout the cord and to establish an anterior-to-posterior phase delay among segmental oscillators. We addressed two questions about this system. First, do the intrasegmental swim circuits in each ganglion function as a single oscillator or do they comprise a pair of coupled oscillators? Second, what are the relative strengths of the ascending and descending intersegmental interactions between the segmental oscillators? Experiments were carried out on semi-intact leeches (Hirudo medicinalis) and on isolated leech nerve cords in which "Z-cut" ganglia were generated by cutting one lateral connective nerve anterior and the contralateral connective nerve posterior to the target ganglion. In these Z-cut ganglia, all rhythmic ascending intersegmental input is conveyed via one lateral connective while rhythmic descending input is conveyed via the contralateral connective. We found that rhythmic bursting recorded from the left and right sides of Z-cut ganglia had identical cycle periods with no phase difference, despite strong intersegmental inputs with differing periods from the two swimming ends of the preparations. We conclude that the swim circuits within individual leech ganglia act as single units. Moreover, we determined through correlation and Fourier spectral analyses, that the functional strengths of ascending and descending intersegmental inputs to Z-cut ganglia located in the middle of the nerve cord are approximately equal. PMID- 11495954 TI - Effect of acute exposure to ammonia on glutamate transport in glial cells isolated from the salamander retina. AB - A rise of brain ammonia level, as occurs in liver failure, initially increases glutamate accumulation in neurons and glial cells. We investigated the effect of acute exposure to ammonia on glutamate transporter currents in whole cell clamped glial cells from the salamander retina. Ammonia potentiated the current evoked by a saturating concentration of L-glutamate, and decreased the apparent affinity of the transporter for glutamate. The potentiation had a Michaelis-Menten dependence on ammonia concentration, with a K(m) of 1.4 mM and a maximum potentiation of 31%. Ammonia also potentiated the transporter current produced by D-aspartate. Potentiation of the glutamate transport current was seen even with glutamine synthetase inhibited, so ammonia does not act by speeding glutamine synthesis, contrary to a suggestion in the literature. The potentiation was unchanged in the absence of Cl(-) ions, showing that it is not an effect on the anion current gated by the glutamate transporter. Ammonium ions were unable to substitute for Na+ in driving glutamate transport. Although they can partially substitute for K+ at the cation counter-transport site of the transporter, their occupancy of these sites would produce a potentiation of < 1%. Ammonium, and the weak bases methylamine and trimethylamine, increased the intracellular pH by similar amounts, and intracellular alkalinization is known to increase glutamate uptake. Methylamine and trimethylamine potentiated the uptake current by the amount expected from the known pH dependence of uptake, but ammonia gave a potentiation that was larger than could be explained by the pH change, and some potentiation of uptake by ammonia was still seen when the internal pH was 8.8, at which pH further alkalinization does not increase uptake. These data suggest that ammonia speeds glutamate uptake both by increasing cytoplasmic pH and by a separate effect on the glutamate transporter. Approximately two-thirds of the speeding is due to the pH change. PMID- 11495955 TI - Preserving and restoring behavioral potential within the spinal cord using an instrumental training paradigm. AB - We have shown that spinal cord neurons can support a simple form of instrumental learning. In a typical experiment, rats are spinalized at the second thoracic vertebra (T(2)) and given shock to one hindleg. One group (master) receives shock whenever the leg is extended. This response-contingent shock causes an increase in response duration that decreases net shock exposure. This instrumental learning is not observed in yoked controls that receive the same amount of shock independent of leg position (noncontingent shock). Interestingly, rats that have received noncontingent shock also fail to learn when they are subsequently exposed to response-contingent shock on either the ipsilateral or contralateral leg. Just 6 min of noncontingent nociceptive stimulation, applied to the leg or tail, undermines behavioral potential for up to 48 h. The present experiments explore whether a behavioral therapy can prevent and/or reverse this deficit. In experiment 1, spinalized rats received 30 min of training with contingent shock, noncontingent shock, or nothing prior to noncontingent tailshock. They were then tested with contingent shock to the contralateral hindleg. Rats that had received noncontingent shock alone failed to learn. Prior exposure to contingent shock had an immunizing effect that prevented the deficit. Experiment 2 examined whether training with contingent shock after noncontingent shock exposure would restore behavioral potential. To facilitate performance during contingent shock training, subjects were given an intrathecal injection of the opioid antagonist naltrexone, a drug treatment that temporarily blocks the expression of the behavioral deficit. Twenty-four hours later subjects were tested with contingent shock on either the ipsilateral or contralateral leg. We found that naltrexone combined with contingent shock therapy restored spinal cord function. Naltrexone alone had no effect. The results suggest that noncontingent nociceptive stimulation can undermine behavioral potential after spinal cord injury and that instrumental training can help preserve, and protect, spinal cord function. PMID- 11495956 TI - Differential projections of thermoreceptive and nociceptive lamina I trigeminothalamic and spinothalamic neurons in the cat. AB - The projections of 40 trigeminothalamic or spinothalamic (TSTT) lamina I neurons were mapped using antidromic activation from a mobile electrode array in barbiturate anesthetized cats. Single units were identified as projection cells from the initial array position and characterized with natural cutaneous stimuli as nociceptive-specific (NS, n = 9), polymodal nociceptive (HPC, n = 8), or thermoreceptive-specific (COOL, n = 22; WARM, n = 1) cells. Thresholds for antidromic activation were measured from each electrode in the mediolateral array at vertical steps of 250 microm over a 7-mm dorsoventral extent in two to eight (median = 6.0) anteroposterior planes. Histological reconstructions showed that the maps encompassed all three of the main lamina I projection targets observed in prior anatomical work, i.e., the ventral aspect of the ventroposterior complex (vVP), the dorsomedial aspect of the ventroposterior medial nucleus (dmVPM), and the submedial nucleus (Sm). The antidromic activation foci were localized to these sites (and occasional projections to other sites were also observed, such as the parafascicular nucleus and zona incerta). The projections of thermoreceptive and nociceptive cells differed. The projections of the thermoreceptive-specific cells were 20/23 to dmVPM, 21/23 to vVP, and 17/23 to Sm, whereas the projections of the NS cells were 1/9 to dmVPM, 9/9 to vVP, and 9/9 to Sm and the projections of the HPC cells were 0/8 to dmVPM, 7/8 to vVP, and 6/8 to Sm. Thus nearly all thermoreceptive cells projected to dmVPM, but almost no nociceptive cells did. Further, thermoreceptive cells projected medially within vVP (including the basal ventral medial nucleus), while nociceptive cells projected both medially and more laterally, and the ascending axons of thermoreceptive cells were concentrated in the medial mesencephalon, while the axons of nociceptive cells ascended in the lateral mesencephalon. These findings provide evidence for anatomical differences between these physiological classes of lamina I cells, and they corroborate prior anatomical localization of the lamina I TSTT projection targets in the cat. These results support evidence indicating that the ventral aspect of the basal ventral medial nucleus is important for thermosensory behavior in cats, consistent with the view that this region is a primordial homologue of the posterior ventral medial nucleus in primates. PMID- 11495957 TI - AMPA and NMDA receptors regulate responses of neurons in the rat's inferior colliculus. AB - The contribution of N-methyl-D-aspartate (NMDA) and AMPA receptors to auditory responses in the rat's inferior colliculus was examined by recording single-unit activity before, during, and after local iontophoretic application of receptor specific antagonists. Tone bursts and sinusoidal amplitude modulated sounds were presented to one ear, and recordings were made from the contralateral central nucleus of inferior colliculus (ICC). The receptor specific antagonists, (+/-)-3 (2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) for NMDA receptors and 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide (NBQX) for AMPA receptors, were released at the recording site through a multi-barreled pipette. For most neurons, either CPP or NBQX alone resulted in a reversible reduction in the number of action potentials evoked by tonal stimulation. For neurons with an onset response pattern, NBQX either completely eliminated or greatly reduced the number of action potentials. CPP also reduced the number of action potentials but had a less pronounced effect than NBQX. For neurons with a sustained firing pattern, NBQX reduced the total number of action potentials, but had a preferential effect on the early part (first 10-20 ms) of the response. CPP also resulted in a reduction in the total number of action potentials, but had a more pronounced effect on the later part (>20 ms) of the response. These results indicate that both AMPA and NMDA receptors contribute to sound evoked excitatory responses in the ICC. They have a selective influence on early and late components of tone-evoked responses. Both receptor types are involved in generating excitatory responses across a wide range of sound pressure levels as indicated by rate level functions obtained before and during drug application. In addition, both CPP and NBQX reduced responses to sinusoidal amplitude modulated sounds. The synchrony of firing to the modulation envelope as measured by vector strength at different rates of modulation was not greatly affected by either CPP or NBQX in spite of the decrease in firing rate. PMID- 11495958 TI - Intracellular correlates of spatial memory acquisition in hippocampal slices: long-term disinhibition of CA1 pyramidal cells. AB - Despite many advances in our understanding of synaptic models of memory such as long-term potentiation and depression, cellular mechanisms that correlate with and may underlie behavioral learning and memory have not yet been conclusively determined. We used multiple intracellular recordings to study learning-specific modifications of intrinsic membrane and synaptic responses of the CA1 pyramidal cells (PCs) in slices of the rat dorsal hippocampus prepared at different stages of the Morris water maze (WM) task acquisition. Schaffer collateral stimulation evoked complex postsynaptic potentials (PSP) consisting of the excitatory and inhibitory postsynaptic potentials (EPSP and IPSP, respectively). After rats had learned the WM task, our major learning-specific findings included reduction of the mean peak amplitude of the IPSPs, delays in the mean peak latencies of the EPSPs and IPSPs, and correlation of the depolarizing-shifted IPSP reversal potentials and reduced IPSP-evoked membrane conductance. In addition, detailed isochronal analyses revealed that amplitudes of both early and late IPSP phases were reduced in a subset of the CA1 PCs after WM training was completed. These reduced IPSPs were significantly correlated with decreased IPSP conductance and with depolarizing-shifted IPSP reversal potentials. Input-output relations and initial rising slopes of the EPSP phase did not indicate learning-related facilitation as compared with the swim and naive controls. Another subset of WM trained CA1 PCs had enhanced amplitudes of action potentials but no learning specific synaptic changes. There were no WM training-specific modifications of other intrinsic membrane properties. These data suggest that long-term disinhibition in a subset of CA1 PCs may facilitate cell discharges that represent and record the spatial location of a hidden platform in a Morris WM. PMID- 11495959 TI - Transmission security for single, hair follicle-related tactile afferent fibers and their target cuneate neurons in cat. AB - Transmission from single, identified hair follicle afferent (HFA) nerve fibers to their target neurons of the cuneate nucleus was examined in anesthetized cats by means of paired recording from individual cuneate neurons and from fine, intact fascicles of the lateral branch of the superficial radial nerve in which it is possible to identify and monitor the activity of each group II fiber. Selective activation of individual HFA fibers was achieved by means of focal vibrotactile skin stimulation. Forearm denervation precluded inputs from sources other than the monitored HFA sensory fiber. Transmission characteristics were analyzed for 21 HFA fiber-cuneate neuron pairs in which activity in the single HFA fiber of each pair reliably evoked spike output from the target neuron at a fixed latency. As the cuneate responses to each HFA impulse often consisted of 2 or 3 spikes, in particular at HFA input rates up to approximately 20 imp/s, the synaptic linkage displayed potent amplification and high-gain transmission, characteristics that were confirmed quantitatively in measures of transmission security and cuneate spike output measures. In response to vibrotactile stimuli, the tight phase locking in the responses of single HFA fibers was well retained in the cuneate responses for vibration frequencies up to approximately 200 Hz. On measures of vector strength, the phase locking declined across the synaptic linkage by no more than approximately 10% at frequencies up to 100 Hz. However, limitations on the impulse rates generated in both the HFA fibers their associated cuneate neurons meant that the impulse patterns could not directly signal information about the vibration frequency above 50-100 Hz. Although single HFA fibers are also known to have secure synaptic linkages with spinocervical tract neurons, it is probable that this linkage lacks the capacity of the HFA-cuneate synapse for conveying precise temporal information, in an impulse pattern code, about the frequency parameter of vibrotactile stimuli. PMID- 11495960 TI - Motion information is spatially localized in a visual working-memory task. AB - We asked if the information about stimulus motion used in a visual working-memory task is localized in space. Monkeys compared the directions of two moving random dot stimuli, sample and test, separated by a temporal delay and reported whether the stimuli moved in the same or in different directions. By presenting the two comparison stimuli in separate locations in the visual field, we determined whether information about stimulus direction was spatially localized during the storage and retrieval/comparison components of the task. Two psychophysical measures of direction discrimination provided nearly identical estimates of the critical spatial separation between sample and test stimuli that lead to a loss in threshold. Direction range thresholds measured with dot stimuli consisting of a range of local directional vectors were affected by spatial separation when a random-motion mask was introduced during the delay into the location of the upcoming test. The selective masking at the test location suggests that the information about the remembered direction was localized and available at that location. Direction difference thresholds, measured with coherently moving random dots, were also affected by separation between the two comparison stimuli. The separation at which performance was affected in both tasks increased with retinal eccentricity in parallel with the increase in receptive-field size in neurons in cortical area MT. The loss with transfer of visual information between different spatial locations suggests a contribution of cortical areas with localized receptive fields to the performance of the memory task. The similarity in the spatial scale of the storage mechanism derived psychophysically and the receptive field size of neurons in area MT suggest that MT neurons are central to this task. PMID- 11495961 TI - Membrane properties of principal neurons of the lateral superior olive. AB - In the lateral superior olive (LSO) the firing rate of principal neurons is a linear function of inter-aural sound intensity difference (IID). The linearity and regularity of the "chopper response" of these neurons have been interpreted as a result of an integration of excitatory ipsilateral and inhibitory contralateral inputs by passive soma-dendritic cable properties. To account for temporal properties of this output, we searched for active time- and voltage dependent nonlinearities in whole cell recordings from a slice preparation of the rat LSO. We found nonlinear current-voltage relations that varied with the membrane holding potential. Repetitive regular firing, supported by voltage oscillations, was evoked by current pulses injected from holding potentials near rest, but the response was reduced to an onset spike of fixed short latency when the pulse was injected from de- or hyperpolarized holding potentials. The onset spike was triggered by a depolarizing transient potential that was supported by T type Ca(2+)-, subthreshold Na(+)-, and hyperpolarization-activated (I(H)) conductances sensitive, respectively, to blockade with Ni2+, tetrodotoxin (TTX), and Cs+. In the hyperpolarized voltage range, the I(H), was largely masked by an inwardly rectifying K+ conductance (I(KIR)) sensitive to blockade with 200 microM Ba2+. In the depolarized range, a variety of K+ conductances, including A currents sensitive to blockade with 4-aminopyridine (4-AP) and additional tetraethylammonium (TEA)-sensitive currents, terminated the transient potential and firing of action potentials, supporting a strong spike-rate adaptation. The "chopper response," a hallmark of LSO principal neuron firing, may depend on the voltage- and time-dependent nonlinearities. These active membrane properties endow the LSO principal neurons with an adaptability that may maintain a stable code for sound direction under changing conditions, for example after partial cochlear hearing loss. PMID- 11495962 TI - Ocular counterroll modulates the preferred direction of saccade-related pontine burst neurons in the monkey. AB - Saccade-related burst neurons in the paramedian pontine reticular formation (PPRF) of the head-restrained monkey provide a phasic velocity signal to extraocular motoneurons for the generation of rapid eye movements. In the superior colliculus (SC), which directly projects to the PPRF, the motor command for conjugate saccades with the head restrained in a roll position is represented in a reference frame in between oculocentric and space-fixed coordinates with a clear bias toward gravity. Here we studied the preferred direction of premotor burst neurons in the PPRF during static head roll to characterize their frame of reference with respect to head and eye position. In 59 neurons (short-lead, burst tonic, and long-lead burst neurons), we found that the preferred direction of eye displacement of these neurons changed, relative to head-fixed landmarks, in the horizontal-vertical plane during static head roll. For the short-lead burst neurons and the burst-tonic group, the change was about one-fourth of the amount of ocular counterroll (OCR) and significantly different from a head-centered representation. In the long-lead burst neurons, the rotation of the preferred direction showed a larger trend of about one-half of OCR. During microelectrical stimulation of the PPRF (9 sites in 2 monkeys), the elicited eye movements rotated with about one-half the amount of OCR. In a simple pulley model of the oculomotor plant, the noncraniocentric reference frame of the PPRF output neurons could be reproduced for recently measured pulley positions, if the pulleys were assumed to rotate as a function of OCR with a gain of 0.5. We conclude that the saccadic displacement signal is transformed from a representation in the SC with a clear bias to gravity to a representation in the PPRF that is closely craniocentric, but rotates with OCR, consistent with current concepts of the oculomotor plant. PMID- 11495963 TI - Variations in photoreceptor response dynamics across the fly retina. AB - Gradients in the spatial properties of retinal cells and their relation to image statistics are well documented. However, less is known of gradients in temporal properties, especially at the level of the photoreceptor for which no account exists. Using light flashes and white-noise-modulated light and current stimuli, we examined the spatial and temporal properties of a single class of photoreceptor (R1-6) within the compound eyes of male blowfly, Calliphora vicina. We find that there is a trend toward higher performance at the front of the eye, both in terms of spatiotemporal resolution and signal-to-noise ratio. The receptive fields of frontal photoreceptors are narrower than those of photoreceptors at the side and back of the eye and response speeds are 20% faster. The signal-to-noise ratio at high frequencies is also greatest at the front of the eye, allowing a 30-40% higher information rate. The power spectra of signals and noise indicate that this elevation of performance results both from shorter responses to individual photons and from a more reliable registration of photon arrival times. These distinctions are characteristic of adaptational changes that normally occur on increasing illumination. However, all photoreceptors were absorbing light at approximately the same mean photon rate during our recordings. We therefore suggest that frontal photoreceptors attain a higher state of light adaptation for a given photon rate. This difference may be achieved by a higher density of (Ca2+ permeable) light-gated channels. Consistent with this hypothesis, membrane-impedance measurements show that frontal photoreceptors have a higher specific conductance than other photoreceptors. This higher conductance provides a better temporal performance but is metabolically expensive. Across the eye, temporal resolution is not proportional to spatial (optical) resolution. Neither is it matched obviously to optic flow. Instead we examine the consequences of an improved temporal resolution in the frontal region for the tracking of small moving targets, a behavior exhibited by male flies. We conclude that the temporal properties of a given class of retinal neuron can vary within a single retina and that this variation may be functionally related to the behavioral requirements of the animal. PMID- 11495964 TI - Gaze anchoring to a pointing target is present during the entire pointing movement and is driven by a non-visual signal. AB - A well-coordinated pattern of eye and hand movements can be observed during goal directed arm movements. Typically, a saccadic eye movement precedes the arm movement, and its occurrence is temporally correlated with the start of the arm movement. Furthermore, the coupling of gaze and aiming movements is also observable after pointing initiation. It has recently been observed that saccades cannot be directed to new target stimuli, away from a pointing target stimulus. Saccades directed to targets presented during the final phase of a pointing movement were delayed until after pointing movement offset ("gaze anchoring"). The present study investigated whether ocular gaze is anchored to a pointing target during the entire pointing movement. In experiment 1, new targets were presented at various times during the duration of a pointing movement, triggered by the kinematics arm moment itself (movement onset, peak acceleration/velocity/deceleration, and offset). Subjects had to make a saccade to the new target as fast as possible while maintaining the pointing movement to the initial target. Saccadic latencies were increased by an amount of time that approximately equaled the remaining pointing time after saccadic target presentation, with the majority of saccades executed after pointing movement offset. The nature of the signal driving gaze stabilization during pointing was investigated in experiment 2. In previous experiments where ocular gaze was anchored to a pointing target, subjects could always see their moving arm, thus it was unknown whether a visual image of the moving arm, an afferent (proprioceptive) signal or an efferent (motor control related) signal produced gaze anchoring. In experiment 2 subjects had to point with or without vision of the moving arm to test whether a visual signal is used to anchor gaze to a pointing target. Results indicate that gaze anchoring was also observed without vision of the moving arm. The findings support the existence of a mechanism enforcing ocular gaze anchoring during the entire duration of a pointing movement. Moreover, such a mechanism uses an internally generated, or proprioceptive, nonvisual signal. Possible neural substrates underlying these processes are discussed, as well as the role of selective attention. PMID- 11495965 TI - Learning to move amid uncertainty. AB - We studied how subjects learned to make movements against unpredictable perturbations. Twelve healthy human subjects made goal-directed reaching movements in the horizontal plane while holding the handle of a two-joint robotic manipulator. The robot generated viscous force fields that perturbed the limb perpendicular to the desired direction of movement. The amplitude (but not the direction) of the viscous field varied randomly from trial to trial. Systems identification techniques were employed to characterize how subjects adapted to these random perturbations. Subject performance was quantified primarily using the peak deviation from a straight-line hand path. Subjects adapted their arm movements to the sequence of random force-field amplitudes. This adaptive response compensated for the approximate mean from the random sequence of perturbations and did not depend on the statistical distribution of that sequence. Subjects did not adapt by directly counteracting the mean field strength itself on each trial but rather by using information about perturbations and movement errors from a limited number of previous trials to adjust motor commands on subsequent trials. This strategy permitted subjects to achieve near optimal performance (defined as minimizing movement errors in a least-squares sense) while maintaining computational efficiency. A simple model using information about movement errors and perturbation amplitudes from a single previous trial predicted subject performance in stochastic environments with a high degree of fidelity and further predicted key performance features observed in nonstochastic environments. This suggests that the neural structures modified during motor adaptation require only short-term memory. Explicit representations regarding movements made more than a few trials in the past are not used in generating optimal motor responses on any given trial. PMID- 11495966 TI - Enhanced responses of spinal dorsal horn neurons to heat and cold stimuli following mild freeze injury to the skin. AB - The effects of a mild freeze injury to the skin on responses of nociceptive dorsal horn neurons to cold and heat stimuli were examined in anesthetized rats. Electrophysiological recordings were obtained from 72 nociceptive spinal neurons located in the superficial and deep dorsal horn. All neurons had receptive fields (RFs) on the glabrous skin of the hindpaw, and neurons were functionally divided into wide dynamic range (WDR) and high-threshold (HT) neurons. Forty-four neurons (61%) were classified as WDR and responded to both innocuous and noxious mechanical stimuli (mean mechanical threshold of 12.8 +/- 1.6 mN). Twenty-eight neurons (39%) were classified as HT and were excited only by noxious mechanical stimuli (mean mechanical threshold of 154.2 +/- 18.3 mN). Neurons were characterized for their sensitivity heat (35 to 51 degrees C) and cold (28 to -12 degrees C) stimuli applied to their RF. Among WDR neurons, 86% were excited by both noxious heat and cold stimuli, while 14% responded only to heat. For HT neurons, 61% responded to heat and cold stimuli, 32% responded only to noxious heat, and 7% responded only to noxious cold. Effects of a mild freeze injury (-15 degrees C applied to the RF for 20 s) on responses to heat and cold stimuli were examined in 30 WDR and 22 HT neurons. Skin freezing was verified as an abrupt increase in skin temperature at the site of injury due to the exothermic reaction associated with crystallization. Freezing produced a decrease in response thresholds to heat and cold stimuli in most WDR and HT neurons. WDR and HT neurons exhibited a mean decrease in response threshold for cold of 9.0 +/- 1.3 degrees C and 10.0 +/- 1.6 degrees C, respectively. Mean response thresholds for heat decreased 4.0 +/- 0.4 degrees C and 4.3 +/- 1.3 degrees C in WDR and HT neurons, respectively. In addition, responses to suprathreshold cold and heat stimuli increased. WDR and HT neurons exhibited an 89% and a 192% increase in response across all cold stimuli, and a 93 and 92% increase in responses evoked across all heat stimuli, respectively. Our results demonstrate that many spinal neurons encode intensity of noxious cold as well as noxious heat over a broad range of stimulus temperatures. Enhanced responses of WDR and HT neurons to cold and heat stimuli after a mild freeze injury is likely to contribute to thermal hyperalgesia following a similar freeze injury in humans. PMID- 11495967 TI - Interactions between ON and OFF signals in directional motion detectors feeding the not of the wallaby. AB - An apparent motion stimulus is used to probe the interactions between signals representing brightness increments (ON stimuli) and decrements (OFF stimuli) in the directional motion detectors forming the input to the nucleus of the optic tract (NOT) of the wallaby, Macropus eugenii. Direction-selective NOT neurons increase their firing rates during image motion from temporal-to-nasal over the contralateral eye (preferred direction) and their spontaneous activities are inhibited by motion in the opposite, anti-preferred direction. An apparent motion stimulus, consisting of neighboring vertical bars, where the brightness can be manipulated independently, also produces directional responses. Preferred direction sequences of brightness changes of like polarities (ON-ON or OFF-OFF) produce increased firing rates while sequences of opposite polarities (ON-OFF or OFF-ON) in the same direction produce relatively small excitatory responses or inhibit the spontaneous rate. For apparent motion in the anti-preferred direction, these directional properties are reversed, showing that signals for brightness increments and decrements provide inputs to the same motion detectors. There is no evidence for segregation of motion detectors into those receiving only half-wave rectified inputs. Interactions between ON and OFF signals utilize the sign of the incoming signals. An array of Reichardt-type motion detectors receiving inputs represented as positive and negative values for ON and OFF stimuli, respectively, are used to simulate the NOT responses. The brightness signals enter band-pass temporal filters prior to motion detection. By altering the time constants of these prefilters, it was possible to accurately simulate the time courses of each cell's responses. PMID- 11495968 TI - Persistent Na+ current and Ca2+ current boost graded depolarization of rat retinal amacrine cells in culture. AB - Retinal amacrine cells are depolarized by the excitatory synaptic input from bipolar cells. When a graded depolarization exceeds the threshold level, trains of action potentials are generated. There have been several reports that both spikes and graded depolarization are sensitive to tetrodotoxin (TTX). In the present study, we investigated the contribution of voltage-gated currents to membrane depolarization by using rat GABAergic amacrine cells in culture recorded by the patch-clamp method. Injection of a negative current induced membrane hyperpolarization, the waveform of which can be well fitted by a single exponential function. Injection of positive current depolarized the cell, and the depolarization exceeded the amplitude expected from the passive properties of the membrane. The boosted depolarization sustained after the current was turned off. Either 1 microM TTX or 2 mM Co2+ suppressed the boosted depolarization, and co application of TTX and Co2+ blocked it completely. Under the voltage clamp, we identified a transient Na+ current (fast I(Na)), a TTX-sensitive persistent current that reversed the polarity near the equilibrium potential of Na+ (I(NaP)), and three types of Ca2+ currents (I(Ca)), L, N, and the pharmacological agent-resistant type (R type). These findings suggest that the I(NaP) and I(Ca) of amacrine cells boost depolarization evoked by the excitatory synaptic input, and they may aid the spread of electrical signals among dendritic arbors of amacrine cells. PMID- 11495969 TI - Broadly tuned spinal neurons for each form of fictive scratching in spinal turtles. AB - Behavioral choice can be mediated either by a small number of sharply tuned neurons or by large populations of broadly tuned neurons. This issue can be conveniently examined in the turtle spinal cord, which generates each of three forms of scratching-rostral, pocket, and caudal-in response to mechanical stimulation in each of three adjacent regions of the body surface. Previous research showed that many propriospinal neurons are broadly tuned to either the rostral scratch region or the pocket scratch region, but responses to caudal scratch stimulation could not be examined in that reduced preparation. In the current study, individual spinal neurons were recorded extracellularly from the gray matter of the turtle spinal cord hindlimb enlargement, while sites in the rostral, pocket, and caudal scratch regions were mechanically stimulated. Many neurons were broadly tuned to the caudal scratch region; other neurons were broadly tuned to either the pocket scratch or rostral scratch region. All three types were typically found within a single animal. These data are consistent with the hypothesis that the turtle spinal cord relies on large populations of broadly tuned neurons to select each of the three forms of scratching. In addition, neurons that were broadly tuned to each of the scratch regions were typically found in each spinal cord segment and within the same range of mediolateral and dorsoventral locations. Providing that these neurons are related to the selection and generation of the three forms of scratching, this would indicate that cells of this type are not segregated into distinct regions of the spinal cord gray matter. PMID- 11495970 TI - Rhythmicity of spinal neurons activated during each form of fictive scratching in spinal turtles. AB - Are behaviors that rely on common muscles and motoneurons generated by separate or overlapping groups of pattern-generating neurons? This question was investigated for the three forms of scratching in immobilized, spinal turtles. Individual neurons were recorded extracellularly from the gray matter through most of the spinal cord hindlimb enlargement gray matter, but were avoided in the region of motoneuron cell bodies. Each form of fictive scratching was elicited by mechanical stimulation of the body surface. The rhythmic modulation of spinal neurons was assessed using phase histograms and circular statistics. The degree of rhythmic modulation and the phase preference of each rhythmically active neuron were measured with respect to the activity cycle of the ipsilateral hip flexor nerve. The action potentials of rhythmic neurons tended to be concentrated in a particular phase of the ipsilateral hip flexor activity cycle no matter which form of fictive scratching was elicited. This consistent phase preference suggests that some of these neurons may contribute to generation of the hip rhythm for all three forms of scratching, strengthening the case that vertebrate pattern-generating circuitry for distinct behaviors can be overlapping. The degree of rhythmic modulation of each unit during fictive scratching was consistently correlated with the dorsoventral location of the recording, but not with the mediolateral or rostrocaudal location; neurons located more ventrally tended to be more rhythmic. The phase preferences of units were related to the region of the body surface to which each neuron responded maximally (i.e., the region to which each unit was broadly tuned). Units tuned to the rostral scratch or pocket scratch region tended to have a phase preference during ipsilateral hip flexor activity, whereas units tuned to the caudal scratch region did not. This suggests the hypothesis that the hip flexes further during rostral and pocket scratching, and extends further during caudal scratching, due to the net effects of a population of spinal interneurons that are both broadly tuned and rhythmically active. PMID- 11495971 TI - Abundant GFP expression and LTP in hippocampal acute slices by in vivo injection of sindbis virus. AB - Virus-mediated gene transfer into neurons is a powerful tool for the analysis of neuronal structure and function. Recombinant sindbis virus has been previously used to study protein function in hippocampal neuron cultures as well as in hippocampal organotypic slice cultures. Nevertheless, some concern still exists about the physiological relevance of these cultured preparations. Acute hippocampal slices are a widely used preparation for the study of synaptic transmission, but currently recombinant gene delivery is usually achieved only through time-consuming transgenic techniques. In this study, we show that a subregion of the CA1 area in acute hippocampal slices can be specifically altered to express a gene of interest. A sindbis virus vector carrying an enhanced green fluorescent protein (EGFP) reporter was injected in vivo into the hippocampus of adult rats. After 18 h, rats were killed, and acute hippocampal slices, infected in the CA1 field, were analyzed morphologically and electrophysiologically. Infected slices showed healthy and stable electrophysiological responses as well as long-term potentiation. In addition, infected pyramidal cells were readily recognized in living slices by two-photon imaging. Specifically, the introduction of an EGFP-Actin fusion protein greatly enhanced the detection of fine processes and dendritic spines. We propose this technique as an efficient tool for studying gene function in adult hippocampal neurons. PMID- 11495972 TI - Listening through different ears alters spatial response fields in ferret primary auditory cortex. AB - The localization of sounds in space is based on spatial cues that arise from the acoustical properties of the head and external ears. Individual differences in localization cue values result from variability in the shape and dimensions of these structures. We have mapped spatial response fields of high-frequency neurons in ferret primary auditory cortex using virtual sound sources based either on the animal's own ears or on the ears of other subjects. For 73% of units, the response fields measured using the animals' own ears differed significantly in shape and/or position from those obtained using spatial cues from another ferret. The observed changes correlated with individual differences in the acoustics. These data are consistent with previous reports showing that humans localize less accurately when listening to virtual sounds from other individuals. Together these findings support the notion that neural mechanisms underlying auditory space perception are calibrated by experience to the properties of the individual. PMID- 11495973 TI - Impedance control and internal model formation when reaching in a randomly varying dynamical environment. AB - We investigated the effects of trial-to-trial, random variation in environmental forces on the motor adaptation of human subjects during reaching. Novel sequences of dynamic environments were applied to subjects' hands by a robot. Subjects reached first in a "mean field" having a constant gain relating force and velocity, then in a "noise field," having a gain that varied randomly between reaches according to a normal distribution with a mean identical to that of the mean field. The unpredictable nature of the noise field did not degrade adaptation as quantified by final kinematic error and rate of adaptation. To achieve this performance, the nervous system used a dual strategy. It increased the impedance of the arm as evidenced by a significant reduction in aftereffect size following removal of the noise field. Simultaneously, it formed an internal model of the mean of the random environment, as evidenced by a minimization of trajectory error on trials for which the noise field gain was close to the mean field gain. We conclude that the human motor system is capable of predicting and compensating for the dynamics of an environment that varies substantially and randomly from trial to trial, while simultaneously increasing the arm's impedance to minimize the consequence of errors in the prediction. PMID- 11495974 TI - Neurosteroids mediate habituation and tonic inhibition in the auditory midbrain. AB - Habituation of the behavioral response to a repetitive stimulus is a well established observation in perceptual studies and is considered a basic form of nonassociative learning. There is also a long history of physiological studies suggesting that central nervous system habituation is mediated by inhibition. At higher levels of the sensory pathways, such inhibition is mainly contributed by GABAa receptor mechanisms. Concepts of modification of synaptic efficacy that apply to excitatory amino acid synaptic transmission do not have direct parallels with these inhibitory synapses: quantal release of GABA rapidly saturates available receptors at a synapse, placing an upper limit on responsiveness to increased transmitter release. However, pharmacological modulation of GABAa receptor efficacy with exogenous agents (e.g., benzodiazepines and beta carbolines) is known to occur through allosteric mechanisms that modulate the effectiveness (positive and negative) of GABA at this receptor. The most potent endogenous modulators are 5alpha-reduced steroids. Production of these steroids was attenuated in adult rats with systemic injection of Finasteride, a competitive substrate for 5alpha-reductase. This treatment was sufficient to block habituation of the evoked midbrain response to repetitive presentation of an acoustic click. This result confirms that simple habituation is due to an increase in active inhibition, the increase being mediated by steroid modulation of the GABAa-receptor. Finasteride treatment also brought about a 23% increase in the evoked response to a click stimulus, suggesting that 5alpha-reduced steroids normally contribute to tonic inhibition in the rat inferior colliculus. PMID- 11495975 TI - Sonometric measurements of motor-neuron-evoked movements of an internal feeding structure (the radula) in Aplysia. AB - In many systems used to study rhythmic motor programs, the structures that generate behavior are at least partially internal. In these systems, it is often difficult to directly monitor neurally evoked movements. As a consequence, although motor programs are relatively well characterized, it is generally less clear how neural activity is translated into functional movements. This is the case for the feeding system of the mollusk Aplysia. Here we used sonomicrometry to monitor neurally evoked movements of the food-grasping organ in Aplysia, the radula. Movements were evoked by intracellular stimulation of motor neurons that innervate radula muscles that have been extensively studied in reduced preparations. Nevertheless our results indicate that the movements and neural control of the radula are more complex than has been assumed. We demonstrate that motor neurons previously characterized as radula openers (B48) and closers (B8, B15, B16) additionally produce other movements. Moreover, we show that the size of the movement evoked by a motor neuron can depend on the preexisting state of the radula. Specifically, the motor neurons B15 and B16 produce large closing movements when the radula is partially open but produce relatively weak closing movements in a preparation at rest. Thus the efficacy of B15 and B16 as radula closers is context dependent. PMID- 11495976 TI - Anesthesia changes frequency tuning of neurons in the rat primary auditory cortex. AB - The vast majority of investigations on central auditory processing so far were conducted under the influence of an anesthetic agent. It remains unclear, however, to what extend even basic response properties of central auditory neurons are influenced by this experimental manipulation. We used a combination of chronic recording in unrestrained animals, computer-controlled randomized acoustic stimulation, and statistical evaluation of responses to directly compare the response characteristics of single neurons in the awake and anesthetized state. Thereby we were able to quantify the effects of pentobarbital/chloral hydrate anesthesia (Equithesin) on rat auditory cortical neurons. During Equithesin anesthesia, only a portion of central neurons were active and some of their basic response properties were changed. Only 29% of the neurons still had a frequency response area. Their tuning sharpness was increased under anesthesia. Most changes are consistent with an enhancement of inhibitory influences during Equithesin anesthesia. Thus when describing response properties of central auditory neurons, the animal's anesthetic state has to be taken into account. PMID- 11495977 TI - lacZ fusions report gene expression, don't they? PMID- 11495978 TI - Candida albicans proteinases: resolving the mystery of a gene family. PMID- 11495979 TI - A GAS-like gene family in the pathogenic fungus Candida glabrata. AB - In fungi, the cell wall plays a major role in host-pathogen interactions. Despite this, little is known about the molecular basis of cell wall assembly in Candida glabrata, which has emerged as the second most common cause of systemic candidosis. A C. glabrata gene family, CgGAS1-3, that shares significant homologies with both the GAS1 gene of Saccharomyces cerevisiae, which is necessary for cell wall assembly, and the pH-regulated genes PHR1 and PHR2 of Candida albicans, which are involved in cell wall assembly and required for virulence, has been cloned. Among the members of this family, CgGAS1-3 display a unique expression pattern. Both CgGAS1 and CgGAS2 are constitutively expressed. In contrast, CgGAS3 transcript was not detectable under any of the assayed conditions. The C. glabrata actin gene, CgACT1, has also been cloned to be used as a meaningful loading control in Northern blots. CgGAS1 and CgGAS2 were deleted by two different methodological approaches. A rapid PCR-based strategy by which gene disruption was achieved with short regions of homology (50 bp) was applied successfully to C. glabrata. DeltaCggas1 or DeltaCggas2 cells demonstrated similar aberrant morphologies, displaying an altered bud morphology and forming floccose aggregates. These phenotypes suggest a role for CgGAS1 and CgGAS2 in cell wall biosynthesis. Further evidence for this hypothesis was obtained by successful functional complementation of a gas1 null mutation in S. cerevisiae with the C. glabrata CgGAS1 or CgGAS2 gene. PMID- 11495980 TI - A highly polymorphic degenerate microsatellite for molecular strain typing of Candida krusei. AB - Simple sequence repeats, due to their high variability, are widely used for molecular epidemiology of pathogenic micro-organisms. However, their usefulness is restricted by their high instability and low information content. Here, a locus, CKTNR, in the fungal pathogen Candida krusei is described which displays considerable sequence, as well as length, heterogeneity. Alleles of this locus, which contains a degenerate trinucleotide repeat, appear to be stable. The CKTNR polymorphism could serve as the basis for a molecular typing system of C. krusei. Furthermore, analysis of the CKTNR allele distribution suggested that C. krusei reproduces mainly clonally. PMID- 11495981 TI - A survey of heterobasidiomycetous yeasts for the presence of the genes homologous to virulence factors of Filobasidiella neoformans, CNLAC1 and CAP59. AB - Among species of the heterobasidiomycetous yeasts, Filobasidiella neoformans is the only serious pathogen that causes fatal infections in both immunocompromised as well as immunocompetent patients. Three phenotypic characteristics, including growth at 37 degrees C, extracellular polysaccharide capsule and laccase activity, of F. neoformans are known to play major roles in the pathogenicity of the fungus. Several CAP genes involved in polysaccharide capsule formation, as well as the CNLAC1 gene encoding a laccase, have previously been cloned and characterized. To analyse the presence of these Cryptococcus neoformans virulence factors in other heterobasidiomycetous yeasts, numerous species of heterobasidiomycetous yeasts were screened for the presence of laccase activity and a polysaccharide capsule. Species exhibiting laccase activity and possessing a glucuronoxylomannan (GXM) capsule were screened for homologues of both the CAP59 gene and the CNLAC1 gene of F. neoformans. Southern blots of genomic DNA from GXM capsule-producing species exhibited no discernible hybridization to the CAP59 DNA sequence except for the two varieties of F. neoformans and Cryptococcus podzolicus. Although discernible, the hybridization band observed with the DNA of C. podzolicus was faint. Oligonucleotide primers constructed using the CAP59 gene sequence also failed to yield PCR products from DNAs of these yeasts except for the two varieties of F. neoformans. These results, coupled with the absence of a CAP59 homologue in the database, suggested the CAP59 gene to be unique to F. neoformans. C. podzolicus was the only species besides F. neoformans that possessed a capsule and expressed strong laccase activity on various media containing phenolic compounds. A CNLAC1 homologue was isolated from C. podzolicus while it was not detected in the species producing beige to faint tan colonies on media with phenolic compounds. Compared to the CNLAC1 sequence of four serotypes of F. neoformans, the CNLAC1 homologue of C. podzolicus showed the highest homology to that of serotype B/C strains and the lowest homology to that of serotype A strains. PMID- 11495982 TI - Adr1 and Cat8 synergistically activate the glucose-regulated alcohol dehydrogenase gene ADH2 of the yeast Saccharomyces cerevisiae. AB - Glucose-repressible alcohol dehydrogenase II, encoded by the ADH2 gene of the yeast Saccharomyces cerevisiae, is transcriptionally controlled by the activator Adr1, binding UAS1 of the control region. However, even in an adr1 null mutant, a substantial level of gene derepression can be detected, arguing for the existence of a further mechanism of activation. Here it is shown that the previously identified UAS2 contains a distantly related variant of the carbon source responsive element (CSRE) initially found upstream of gluconeogenic genes. In a mutant defective for the CSRE-binding factor Cat8, derepression of an ADH2-lacZ fusion was reduced to about 12% of the wild-type level. Gene expression in a cat8 adr1 double mutant decreased almost to the basal level of the glucose-repressed promoter. CSRE(ADH2) present in a single copy turned out to be a weak UAS element, while a significant synergism of gene activation was found in the presence of at least two copies. Its importance for regulated gene activation was confirmed by site-directed mutagenesis of the CSRE in the natural ADH2 control region. Direct binding of Cat8 to CSRE(ADH2) could be shown by electrophoretic retardation of the corresponding protein/DNA complex in the presence of a specific antibody. In contrast to what was shown previously for CSRE sequence variants, no significant influence of the isofunctional activator Sip4 on CSRE(ADH2) was detected. In conclusion, these results show a derepression of ADH2 by synergistically acting regulators Adr1 (interacting with UAS1) and Cat8, binding to UAS2 (=CSRE(ADH2)). PMID- 11495983 TI - Lysine aminopeptidase of Aspergillus niger. AB - Conserved regions within the M1 family of metallo-aminopeptidases have been used to clone a zinc aminopeptidase from the industrially used fungus Aspergillus niger. The derived amino acid sequence of ApsA is highly similar to two yeast zinc aminopeptidases, LAPI and AAPI (53.3 and 50.9% overall similarity, respectively), two members of the M1 family of metallo-aminopeptidases. The encoding gene was successfully overexpressed in A. niger and the overexpressed product was purified and characterized. Aminopeptidase A was found to be active towards a number of amino acid p-nitroanilide (pNA) substrates, viz. K-pNA, R pNA, L-pNA, M-pNA, A-pNA and F-pNA. The most preferred N-terminal amino acid is lysine and not leucine, arginine or alanine, the N-terminal amino acids preferred by the yeast homologues. The K(m) and K(cat) for K-pNA and L-pNA were 0.17 mM and 0.49 microkat mg(-1), and 0.16 mM and 0.31 microkat mg(-1), respectively. The pH optimum of the enzyme is between 7.5 and 8, whereas the enzyme is stable between pH 5 and 8. The enzyme is inhibited by the metal chelators EGTA, EDTA and 1,10 phenanthrolin. Bestatin was also able to inhibit the activity. PMID- 11495984 TI - Resolvase-like recombination performed by the TP901-1 integrase. AB - The site-specific recombination system of temperate lactococcal bacteriophage TP901-1 is unusual in several respects. First, the integrase belongs to the family of extended resolvases rather than to the lambda integrase family and second, in the presence of this integrase, a 56 bp attP fragment is sufficient for efficient recombination with the chromosomal attB site in the host Lactococcus lactis subsp. cremoris MG1363. In the present work, this attB site was analysed and a 43 bp attB region was found to be the smallest fragment able to participate fully in recombination. In vitro studies showed that the TP901-1 integrase binds this 43 bp attB fragment, the 56 bp attP and a larger attP fragment with equal affinity. Mutational analysis of the 5 bp common core region (TCAAT) showed that the TC dinucleotide is essential for recombination, but not for binding of the integrase, whereas none of the last three bases are important for recombination. When a number of attL sites, obtained by recombination between an attB site containing a mutation in this TC dinucleotide and a wild-type attP site, were sequenced, a mix of sites with the wild-type or the mutated sequence was obtained. These results are consistent with the hypothesis that the TC dinucleotide constitutes the TP901-1 overlap region. A 2 bp overlap region has been observed in recombination reactions catalysed by all other members of the resolvase/invertase family tested so far. By selecting for attB sites with a decreased ability to participate in recombination, two bases located outside the core region of attB were shown to be involved in the in vitro binding of the TP901-1 integrase. PMID- 11495985 TI - Development of improved versatile broad-host-range vectors for use in methylotrophs and other Gram-negative bacteria. AB - Full exploitation of the information available in bacterial genome sequences requires the availability of facile tools for rapid genetic manipulation. One bacterium for which new genetic tools are needed is the methylotroph Methylobacterium extorquens AM1. IncQ and small IncP vectors were shown to be unsuitable for use in this bacterium, but a spontaneous mutant of a small IncP plasmid was isolated that functioned efficiently in M. extorquens AM1. This plasmid was sequenced and used as a base for developing improved broad-host-range cloning vectors. These vectors were found to replicate in a wide variety of bacterial species and have the following advantages: (1) high copy number in Escherichia coli; (2) small size (7.2 and 8.0 kb); (3) complete sequences; (4) variety of unique restriction sites; (5) blue-white screening via lacZalpha; (6) conjugative mobilization between bacterial species; and (7) readily adaptable into species-specific promoter-probe and expression vectors. Two low-background promoter-probe vectors were constructed based on these cloning vectors with either lacZ or xylE as reporter genes; these were shown to report gene expression effectively in M. extorquens AM1. Specific expression vectors were developed for use in M. extorquens AM1, which were shown to express foreign genes at significant levels, and a simple strategy is outlined to develop specific expression vectors for other bacteria. The strong mxaF promoter was used for expression, since E. coli lac-derived promoters were expressed at very low levels. This suite of genetic tools will enable a more sophisticated analysis of the physiology of M. extorquens AM1, and these vectors should also be valuable tools in the study of a variety of bacterial species. PMID- 11495986 TI - Gene replacement in cyanobacteria mediated by a dominant streptomycin-sensitive rps12 gene that allows selection of mutants free from drug resistance markers. AB - Chromosomal gene replacement in cyanobacteria often relies upon the availability of drug resistance markers, and thus multiple replacements have been restricted. Here, a versatile gene replacement system without this restriction is reported in a unicellular cyanobacterium, Synechococcus sp. PCC 7942. The system is based upon the dominance of a streptomycin-sensitive rps12 gene encoding a ribosomal S12 protein over a streptomycin-resistant rps12-R43 allele with a Lys-43-->Arg substitution. To demonstrate the utility of this method, a cassette consisting of the wild-type rps12 gene and a kan gene conferring kanamycin resistance was integrated into the rps12-R43 mutant at the psbAI locus encoding photosystem II D1 protein, resulting in streptomycin-sensitive merodiploids. Despite spontaneous gene conversion in these merodiploids to produce streptomycin-resistant progeny at frequencies ranging from 1x10(-5) to 5x10(-5), homologous recombination could be induced by transforming the merodiploids with template plasmids carrying psbAI 5' and 3' non-coding sequences flanking the D1 coding sequence, which was then replaced by either the gfp ORF for a green fluorescent protein or a precise deletion. Depending on the replication ability of the template plasmids, at most 3-16% of streptomycin-resistant progeny of the merodiploids after transformation were homogenote recombinants with concomitant loss of the kan gene, even in these polyploid cyanobacteria. The rps12-mediated gene replacement thus makes it possible to construct mutants free from drug resistance markers and opens a way to create cyanobacterial strains bearing an unlimited number of gene replacements. PMID- 11495987 TI - Endogenous isolation of replicon probes for assessing plasmid ecology of marine sediment microbial communities. AB - Six functional replication origins (repGA14, repGA33, repGA70, repSD41, repSD164 and repSD172), obtained from endogenously isolated, broad-host-range (BHR) marine plasmids ranging in size from 5 to 60 kb, were used to determine plasmid occurrence in three coastal marine sediment sites (in California, Georgia and South Carolina, USA). The plasmid-specific replicons were isolated from plasmid bearing marine sediment bacteria belonging to the alpha and gamma subclasses of the Proteobacteria. The plasmid sources of the endogenous replicons were considered to be cryptic due to a lack of identifiable phenotypic traits. The putative Rep proteins from a number of these replicons showed similarity to replicons of two recognized families: RCR group III (repSD164) and the FIA family of theta group A (repSD41, repSD121, repGA33 and repGA14). Plasmids isolated from marine bacteria belonging to the genera Pseudoalteromonas, Shewanella and Vibrio cultivated from geographically different coastal sites exhibited homology to two of the marine plasmid replicons, repSD41 and repGA70, obtained from a Vibrio sp. The repGA33 plasmid origin, obtained from a Shewanella sp. isolated from coastal Georgia, was detected in 7% of the Georgia marine sediment Shewanella sp. isolates. Microbial community DNA extracted from marine sediments was also screened for the presence of the plasmid replication sequences. Community DNA samples amplified by PCR yielded a positive signal for the repSD172 and repGA14 replication sequences. The replication origin of BHR plasmid RK2 (IncP) was also detected in marine Vibrio sp. and microbial community DNA extracted from the three coastal sites. These findings provide molecular evidence that marine sediment bacteria harbour an untapped population of BHR plasmids. PMID- 11495988 TI - The pdx genetic marker adjacent to the chloramphenicol biosynthesis gene cluster in Streptomyces venezuelae ISP5230: functional characterization. AB - The pdx-4 mutation in Streptomyces venezuelae ISP5230 confers a growth requirement for pyridoxal (pdx) and is a marker for the genetically mapped cluster of genes associated with chloramphenicol biosynthesis. A gene regulating salvage synthesis of vitamin B6 cofactors in S. venezuelae was cloned by transforming a pdx-4 mutant host with the plasmid vector pDQ101 carrying a library of wild-type genomic DNA fragments, and by selecting for complementation of the host's pdx requirement. However, the corresponding replicative plasmid could not be isolated. Southern hybridizations and transduction analysis indicated that the complementing plasmid had integrated into the chromosome; after excision by a second crossover, the plasmid failed to propagate. To avoid loss of the recombinant vector, a pdx-dependent Streptomyces lividans mutant, KAA1, with a phenotype matching that of S. venezuelae pdx-4, was isolated for use as the cloning host. Introduction of pIJ702 carrying an S. venezuelae genomic library into S. lividans KAA1, and selection of prototrophic transformants, led to the isolation of a stable recombinant vector containing a 2.5 kb S. venezuelae DNA fragment that complemented requirements for pdx in both S. venezuelae and S. lividans mutants. Sequence analysis of the cloned DNA located an intact ORF with a deduced amino acid sequence that, in its central and C-terminal regions resembled type-I aminotransferases. The N-terminal region of the cloned DNA fragment aligned closely with distinctive helix-turn-helix motifs found near the N termini of GntR family transcriptional regulators. The overall deduced amino acid sequence of the cloned DNA showed 73% end-to-end identity to a putative GntR type regulator cloned in cosmid 6D7 from the Streptomyces coelicolor A3(2) genome. This location is close to that of pdxA, the first pdx marker in S. coelicolor A3(2) identified and mapped genetically in Sir David Hopwood's laboratory. The S. venezuelae gene and S. coelicolor pdxA are postulated to be homologues regulating vitamin B6 coenzyme synthesis from pdx. PMID- 11495989 TI - p-Aminobenzoic acid and chloramphenicol biosynthesis in Streptomyces venezuelae: gene sets for a key enzyme, 4-amino-4-deoxychorismate synthase. AB - Amplification of sequences from Streptomyces venezuelae ISP5230 genomic DNA using PCR with primers based on conserved prokaryotic pabB sequences gave two main products. One matched pabAB, a locus previously identified in S. venezuelae. The second closely resembled the conserved pabB sequence consensus and hybridized with a 3.8 kb NcoI fragment of S. venezuelae ISP5230 genomic DNA. Cloning and sequence analysis of the 3.8 kb fragment detected three ORFs, and their deduced amino acid sequences were used in BLAST searches of the GenBank database. The ORF1 product was similar to PabB in other bacteria and to the PabB domain encoded by S. venezuelae pabAB. The ORF2 product resembled PabA of other bacteria. ORF3 was incomplete; its deduced partial amino acid sequence placed it in the MocR group of GntR-type transcriptional regulators. Introducing vectors containing the 3.8 kb NcoI fragment of S. venezuelae DNA into pabA and pabB mutants of Escherichia coli, or into the Streptomyces lividans pab mutant JG10, enhanced sulfanilamide resistance in the host strains. The increased resistance was attributed to expression of the pair of discrete translationally coupled p aminobenzoic acid biosynthesis genes (designated pabB/pabA) cloned in the 3.8 kb fragment. These represent a second set of genes encoding 4-amino-4 deoxychorismate synthase in S. venezuelae ISP5230. In contrast to the fused pabAB set previously isolated from this species, they do not participate in chloramphenicol biosynthesis, but like pabAB they can be disrupted without affecting growth on minimal medium. The gene disruption results suggest that S. venezuelae may have a third set of genes encoding PABA synthase. PMID- 11495990 TI - Quorum-sensing-dependent regulation of biosynthesis of the polyketide antibiotic mupirocin in Pseudomonas fluorescens NCIMB 10586. AB - Mupirocin (pseudomonic acid) is a polyketide antibiotic, targeting isoleucyl-tRNA synthase, and produced by Pseudomonas fluorescens NCIMB 10586. It is used clinically as a topical treatment for staphylococcal infections, particularly in contexts where there is a problem with methicillin-resistant Staphylococcus aureus (MRSA). In studying the mupirocin biosynthetic cluster the authors identified two putative regulatory genes, mupR and mupI, whose predicted amino acid sequences showed significant identity to proteins involved in quorum-sensing dependent regulatory systems such as LasR/LuxR (transcriptional activators) and LasI/LuxI (synthases for N-acylhomoserine lactones--AHLs--that activate LasR/LuxR). Inactivation by deletion mutations using a suicide vector strategy confirmed the requirement for both genes in mupirocin biosynthesis. Cross-feeding experiments between bacterial strains as well as solvent extraction showed that, as predicted, wild-type P. fluorescens NCIMB 10586 produces a diffusible substance that overcomes the defect of a mupI mutant. Use of biosensor strains showed that the MupI product can activate the Pseudomonas aeruginosa lasRlasI system and that P. aeruginosa produces one or more compounds that can replace the MupI product. Insertion of a xylE reporter gene into mupA, the first ORF of the mupirocin biosynthetic operon, showed that together mupR/mupI control expression of the operon in such a way that the cluster is switched on late in exponential phase and in stationary phase. PMID- 11495991 TI - Molecular characterization of a deletion/duplication rearrangement in tfd genes from Ralstonia eutropha JMP134(pJP4) that improves growth on 3-chlorobenzoic acid but abolishes growth on 2,4-dichlorophenoxyacetic acid. AB - Ralstonia eutropha JMP134(pJP4) is able to grow on minimal media containing the pollutants 3-chlorobenzoate (3-CB) or 2,4-dichlorophenoxyacetate (2,4-D). tfd genes from the 88 kb plasmid pJP4 encode enzymes involved in the degradation of these compounds. During growth of strain JMP134 in liquid medium containing 3-CB, a derivative strain harbouring a approximately 95 kb plasmid was isolated. This derivative, designated JMP134(pJP4-F3), had an improved ability to grow on 3-CB, but had lost the ability to grow on 2,4-D. Sequence analysis of pJP4-F3 indicated that the plasmid had undergone a deletion of approximately 16 kb, which included the tfdA-tfdS intergenic region, spanning the tfdA gene to a previously unreported IS1071 element. The loss of the tfdA gene explains the failure of the derivative to grow on 2,4-D. A approximately 23 kb duplication of the region spanning tfdR-tfdD(II)C(II)E(II)F(II)-tfdB(II)-tfdK-ISJP4-tfdT tfdC(I)D(I)E(I)F(I)-tfdB(I), giving rise to a 51-kb-long inverted repeat, was also observed. The increase in gene copy number for the tfdCD(DC)EF gene cluster may provide an explanation for the derivative strain's improved growth on 3-CB. These observations are additional examples of the metabolic plasticity of R. eutropha JMP134, one of the more versatile pollutant-degrading bacteria. PMID- 11495992 TI - Growth medium composition-determined regulatory mechanisms are superimposed on CatR-mediated transcription from the pheBA and catBCA promoters in Pseudomonas putida. AB - Expression of the phenol degradation pathway in Pseudomonas putida strain PaW85 requires coordinated transcription of the plasmid-borne pheBA operon encoding catechol 1,2-dioxygenase and phenol monooxygenase, respectively, and the chromosomally encoded catechol degradation catBCA operon. Transcriptional activation from the pheBA and catBCA promoters is regulated by CatR and the catechol degradation pathway intermediate cis,cis-muconate. Here it is shown that physiological control mechanisms are superimposed on this regulatory system. Transcriptional activation from the pheBA and catBCA promoters is growth-phase regulated in P. putida cells grown on rich medium (LB medium). CatR-mediated transcription from these promoters is silenced on rich medium until the transition from exponential to stationary phase. A slight positive effect (threefold) of stationary-phase-specific sigma factor sigma(S) on transcription from the pheBA promoter was observed. Expression of the catBCA promoter was not influenced by the activity of this sigma factor. In contrast to rich growth medium, transcription from the pheBA and catBCA promoters in minimal medium containing a mixture of glucose and sodium benzoate was rapidly induced in exponential culture. It was shown that the presence of amino acids in the culture medium causes exponential silencing of the pheBA and catBCA promoters. The possibility that a hypothetical repressor protein could be involved in physiological control of transcription from the pheBA and catBCA promoters is discussed. PMID- 11495993 TI - Comamonas testosteroni BR6020 possesses a single genetic locus for extradiol cleavage of protocatechuate. AB - A key intermediate for biodegradation of various distinct aromatic growth substrates in Comamonas testosteroni is protocatechuate (Pca), which is metabolized by the 4,5-extradiol (meta) ring fission pathway. A locus harbouring genes from C. testosteroni BR6020 was cloned, dubbed pmd, which encodes the enzymes that degrade Pca. The identity of pmdAB, encoding respectively the alpha- and beta-subunit of the Pca ring-cleavage enzyme, was confirmed by N-terminal sequencing and molecular mass determination of both subunits from the separated enzyme. Disruption of pmdA resulted in a strain unable to grow on Pca and a variety of aromatic substrates funnelled through this compound (m- and p hydroxybenzoate, p-sulfobenzoate, phthalate, isophthalate, terephthalate, vanillate, isovanillate and veratrate). Growth on benzoate and o-aminobenzoate (anthranilate) was not affected in this strain, indicating that these substrates are metabolized via a different lower pathway. Tentative functions for the products of other pmd genes were assigned based on sequence identity and/or similarity to proteins from other proteobacteria involved in uptake or metabolism of aromatic compounds. This study provides evidence for a single lower pathway in C. testosteroni for metabolism of Pca, which is generated by different upper pathways acting on a variety of aromatic substrates. PMID- 11495994 TI - Analysis of transcription of the bph locus of Burkholderia sp. strain LB400 and evidence that the ORF0 gene product acts as a regulator of the bphA1 promoter. AB - Although gene clusters for the degradation of biphenyls and polychlorobiphenyls have been extensively characterized, comparatively little is known about the regulation of their expression. In the present work, different aspects of transcription of the bph locus of the potent polychlorobiphenyl degrader Burkholderia sp. strain LB400 were investigated. An RNA blot analysis of the entire gene cluster revealed that the transcription of all genes encoding biphenyl catabolic enzymes responded similarly to the presence of biphenyl, succinate or a mixture of the two. One region of the locus, encompassing ORF0, was separately transcribed and differently regulated. A single start position was mapped for this monocistronic transcript. Synthesis of the adjacent RNA, encoding subunits of biphenyl dioxygenase, was strongly biphenyl-inducible. In this case, four major 5'-ends were mapped between 25 and 70 bp upstream of the start codon of gene bphA1. Sequence elements between approximately positions 710 and 1080 upstream were required in cis for full functioning of the respective promoter(s) (P(bphA1)). ORF0(-) mutants of strain LB400 retained the ability to grow on biphenyl, but showed decreased concentrations of bphA1A2 RNA and decreased lacZ expression in strains harbouring a reporter system with a bphA1-lacZ transcriptional fusion. This effect was compensated by the introduction of an intact ORF0 in trans, indicating that the ORF0 gene product mediates activation of P(bphA1). PMID- 11495995 TI - Unusual location of two nearby pairs of upstream activating sequences for HbpR, the main regulatory protein for the 2-hydroxybiphenyl degradation pathway of "Pseudomonas azelaica" HBP1. AB - "Pseudomonas azelaica" HBP1 degrades 2-hydroxybiphenyl (2-HBP) and 2,2'-diHBP by employing a meta-cleavage pathway encoded by the hbpCAD genes. The regulatory gene hbpR, located directly upstream of the hbpCAD genes and oriented in the opposite direction, encodes a transcription activator protein belonging to the so called XylR/DmpR subclass within the NtrC family. HbpR activates transcription from two separate sigma(54)-dependent promoters upstream of the hbpC and the hbpD genes, in the presence of the pathway substrates 2-HBP and 2,2'-diHBP. The DNA region upstream of the hbpC gene displays an unusual organization, containing two adjacent 0.3 kb regions that share 71% sequence identity. The DNA region most proximal to the hbpC promoter harbours one pair of putative upstream activating sequences (UASs C-1/C-2) and a small cryptic ORF that shows homology to hbpR itself. The second, more distal, region contains a second pair of putative UASs (UASs C-3/4) and the 5'-part of the hbpR gene. Transcriptional fusions in Escherichia coli between different deletions of the hbpR-hbpC intergenic region and the genes for bacterial luciferase revealed that most if not all of the transcriptional output from the hbpC promoter is mediated from the proximal UASs C-1/C-2. However, when the UASs C-1/C-2 were deleted and UASs C-3/C-4 were placed in an appropriate position with respect to the promoter region, the hbpC promoter was still inducible with 2-HBP, albeit at a lower level. Transcription studies in E. coli and "P. azelaica" revealed that the divergently oriented hbpR gene is expressed constitutively from a sigma(70)-dependent promoter situated within the cryptic ORF. The presence of UAS pair C-3/C-4 mediated a slightly higher promoter activity for transcription of hbpR. PMID- 11495996 TI - Cloning, sequencing and mutagenesis of the genes for aromatic amine dehydrogenase from Alcaligenes faecalis and evolution of amine dehydrogenases. AB - The nucleotide sequence of the aromatic amine utilization (aau) gene region from Alcaligenes faecalis contained nine genes (orf-1, aauBEDA, orf-2, orf-3, orf-4 and hemE) transcribed in the same direction. The aauB and aauA genes encode the periplasmic aromatic amine dehydrogenase (AADH) large and small subunit polypeptides, respectively, and were homologous to mauB and mauA, the genes for the large and small subunits of methylamine dehydrogenase (MADH). aauE and aauD are homologous to mauE and mauD and apparently carry out the same function of transport and folding of the small subunit polypeptide in the periplasm. No analogues of the mauF, mauG, mauL, mauM and mauN genes responsible for biosynthesis of tryptophan tryptophylquinone (the prosthetic group of amine dehydrogenases) were found in the aau cluster. orf-2 was predicted to encode a small periplasmic monohaem c-type cytochrome. No biological function can be assigned to polypeptides encoded by orf-1, orf-3 and orf-4 and mutations in these genes appeared to be lethal. Mutants generated by insertions into mauD were not able to use phenylethylamine, tyramine and tryptamine as a source of carbon and phenylethylamine, 3'-hydroxytyramine (dopamine) and tyramine as a source of nitrogen, indicating that AADH is the only enzyme involved in utilization of primary amines in A. faecalis. AADH genes are present in Alcaligenes xylosoxydans subsp. xylosoxydans, but not in other beta- and gamma-proteobacteria. Phylogenetic analysis of amine dehydrogenases (MADH and AADH) indicated that AADH and MADH evolutionarily diverged before separation of proteobacteria into existing subclasses. PMID- 11495997 TI - The methylcitric acid pathway in Ralstonia eutropha: new genes identified involved in propionate metabolism. AB - From Ralstonia eutropha HF39 null-allele mutants were created by Tn5 mutagenesis and by homologous recombination which were impaired in growth on propionic acid and levulinic acid. From the molecular, physiological and enzymic analysis of these mutants it was concluded that in this bacterium propionic acid is metabolized via the methylcitric acid pathway. The genes encoding enzymes of this pathway are organized in a cluster in the order prpR, prpB, prpC, acnM, ORF5 and prpD, with prpR transcribed divergently from the other genes. (i) prpC encodes a 2-methylcitric acid synthase (42720 Da) as shown by the measurement of the respective enzyme activity, complementation of a prpC mutant of Salmonella enterica serovar Typhimurium and high sequence similarity. (ii) For the translational product of acnM the function of a 2-methyl-cis-aconitic acid hydratase (94726 Da) is proposed. This protein and also the ORF5 translational product are essential for growth on propionic acid, as revealed by the propionic acid-negative phenotype of Tn5-insertion mutants, and are required for the conversion of 2-methylcitric acid into 2-methylisocitric acid as shown by the accumulation of the latter, which could be purified as its calcium salt from the supernatants of these mutants. In contrast, inactivation of prpD did not block the ability of the cell to use propionic acid as carbon and energy source, as shown by the propionic acid phenotype of a null-allele mutant. It is therefore unlikely that prpD from R. eutropha encodes a 2-methyl-cis-aconitic acid dehydratase as proposed recently for the homologous prpD gene from S. enterica. (iii) The translational product of prpB encodes 2-methylisocitric acid lyase (32314 Da) as revealed by measurement of the respective enzyme activity and by demonstrating accumulation of methylisocitric acid in the supernatant of a prpB null-allele mutant. (iv) The expression of prpC and probably also of the other enzymes is regulated and is induced during cultivation on propionic acid or levulinic acid. The putative translational product of prpR (70895 Da) exhibited high similarities to PrpR of Escherichia coli and S. enterica, and might represent a transcriptional activator of the sigma-54 family involved in the regulation of the other prp genes. Since the prp locus of R. eutropha was very different from those of E. coli and S. enterica, an extensive comparison of prp loci available from databases and literature was done, revealing two different classes of prp loci. PMID- 11495998 TI - GcvR interacts with GcvA to inhibit activation of the Escherichia coli glycine cleavage operon. AB - The Escherichia coli glycine cleavage enzyme system, encoded by the gcvTHP operon, catalyses the oxidative cleavage of glycine to CO(2), NH(3) and a one carbon methylene group. Transcription of the gcv operon is positively regulated by GcvA and negatively regulated by GcvA and GcvR. Using a LexA-based system for analysing protein heterodimerization, it is shown that GcvR interacts directly with GcvA in vivo to repress gcvTHP expression. Several mutations in either gcvA or gcvR that result in a loss of gcv repression also result in a loss of GcvA/GcvR heterodimerization. Finally, it is shown that the C-terminal half of GcvA is involved in its interaction with GcvR, whilst the entire GcvR protein appears to be necessary for heterodimerization. PMID- 11495999 TI - Lipoamide dehydrogenase from Corynebacterium glutamicum: molecular and physiological analysis of the lpd gene and characterization of the enzyme. AB - Lipoamide dehydrogenase (LPD) is an essential component of the pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes, both playing a crucial role within the central metabolism of aerobic organisms. Using oligonucleotides designed according to conserved regions of LPD amino acid sequences from several organisms, the lpd gene from Corynebacterium glutamicum was identified and subsequently subcloned. The cloned lpd gene expressed in C. glutamicum cells harbouring the gene on a plasmid showed a 12-fold higher specific LPD activity when compared to the wild-type strain. DNA sequence analysis of a 4524 bp segment containing the lpd gene and adjacent regions revealed that the lpd gene is not flanked by genes encoding other subunits of the pyruvate or 2-oxoglutarate dehydrogenase complexes and predicted an LPD polypeptide of 469 amino acids with an M(r) of 50619. The amino acid sequence of this polypeptide shows between 26 and 58% identity when compared to LPD enzymes from other organisms. Transcriptional analyses revealed that the lpd gene from C. glutamicum is monocistronic (1.45 kb mRNA) and that its transcription is initiated exactly at the nucleotide defined as the translational start. LPD was purified and biochemically characterized. This analysis revealed that the enzyme catalyses the reversible reoxidation of dihydrolipoic acid and NADH:NAD(+) transhydrogenation, and is able to transfer electrons from NADH to various redox-active compounds and quinones. An in vivo participation of C. glutamicum LPD in facilitation of quinone redox cycling is proposed. PMID- 11496000 TI - Azorhizobium caulinodans pyruvate dehydrogenase activity is dispensable for aerobic but required for microaerobic growth. AB - Azorhizobium caulinodans mutant 62004 carries a null allele of pdhB, encoding the E1beta subunit of pyruvate dehydrogenase, which converts pyruvate to acetyl-CoA. This pdhB mutant completely lacks pyruvate oxidation activities yet grows aerobically on C(4) dicarboxylates (succinate, L-malate) as sole energy source, albeit slowly, and displays pleiotropic growth defects consistent with physiological acetyl-CoA limitation. Temperature-sensitive (ts), conditional lethal derivatives of the pdhB mutant lack (methyl)malonate semialdehyde dehydrogenase activity, which thus also allows L-malate conversion to acetyl-CoA. The pdhB mutant remains able to fix N(2) in aerobic culture, but is unable to fix N(2) in symbiosis with host Sesbania rostrata plants and cannot grow microaerobically. In culture, A. caulinodans wild-type can use acetate, beta-D hydroxybutyrate and nicotinate--all direct precursors of acetyl-CoA--as sole C and energy source for aerobic, but not microaerobic growth. Paradoxically, acetyl CoA is thus a required intermediate for microaerobic oxidative energy transduction while not itself oxidized. Accordingly, A. caulinodans energy transduction under aerobic and microaerobic conditions is qualitatively different. PMID- 11496001 TI - Ammonia oxidation by Nitrosomonas eutropha with NO(2) as oxidant is not inhibited by acetylene. AB - The effect of acetylene ((14)C(2)H(2)) on aerobic and anaerobic ammonia oxidation by Nitrosomonas eutropha was investigated. Ammonia monooxygenase (AMO) was inhibited and a 27 kDa polypeptide (AmoA) was labelled during aerobic ammonia oxidation. In contrast, anaerobic, NO(2)-dependent ammonia oxidation (NO(2)/N(2)O(4) as oxidant) was not affected by acetylene. Further studies gave evidence that the inhibition as well as the labelling reaction were O(2) dependent. Cells pretreated with acetylene under oxic conditions were unable to oxidize ammonia with O(2) as oxidant. After these cell suspensions were supplemented with gaseous NO(2), ammonia oxidation activity of about 140 micromol NH(4)(+) (g protein)(-1) h(-1) was detectable under both oxic and anoxic conditions. A significantly reduced acetylene inhibition of the ammonia oxidation activity was observed for cells incubated in the presence of NO. This suggests that NO and acetylene compete for the same binding site on AMO. On the basis of these results a new hypothetical model of ammonia oxidation by N. eutropha was developed. PMID- 11496002 TI - Alteration in cellular fatty acid composition as a response to salt, acid, oxidative and thermal stresses in Lactobacillus helveticus. AB - The fundamental question in this study is concerned with whether the increase of unsaturated fatty acids in the cell membrane is a general response of certain thermotolerant strains or species when exposed to superoptimal temperatures, and in combination with other stresses, especially oxidative stress. A strain of Lactobacillus helveticus, a species widely used as a starter in the dairy industry and able to tolerate high temperature and NaCl concentrations as well as acidic conditions, was chosen for this study. Cells of strain CNBL 1156, grown in its natural medium (i.e. milk whey), were exposed for 100 min to sublethal combinations of temperature, NaCl, H(2)O(2) and pH, modulated according to a Central Composite Design. The fatty acid composition of cell lipid extract was identified by GC/MS. Polynomial equations, able to describe the individual interactive and quadratic effects of the independent variables on cell fatty acid composition, were obtained. The results and the mathematical models relative to the individual fatty acids indirectly suggest that desaturase activation or hyperinduction play an important role in the response to heat stress. In fact, the relative proportions of oleic, linoleic and palmitic acids increased with temperature in a range between 38 and 54 degrees C. The fatty acid profiles included vernolic acid (up to 37% of total fatty acids), an epoxide of linoleic acid not previously reported in microbial cells. In particular, this epoxide was present in cells exposed to low pH in combination with high temperatures and oxidative stress. In conclusion, these results provide experimental support to the hypothesis that the increase of an oxygen-consuming desaturase system, with a consequent increase in fatty acid desaturation, is a cellular response to environmental stresses able to protect the cells of this anaerobic micro-organism from toxic oxygen species and high temperatures. PMID- 11496003 TI - SigB, an alternative sigma factor of the myxobacterium Stigmatella aurantiaca, is synthesized during development and heat shock. AB - Alternative sigma factors have been detected in the myxobacterium Stigmatella aurantiaca during indole-induced sporulation, fruiting body formation and heat shock using an antiserum raised against sigma factor SigB. The time course of sigB gene expression was analysed by RT-PCR and by determining beta-galactosidase activity during development in a merodiploid strain that harboured a sigB-lacZ fusion gene. Inactivation of the sigB gene by insertion of the neo gene resulted in the loss of one sigma factor as shown by Western analysis. Neither fruiting body formation nor sporulation, nor the production of possible SigB targets, such as DnaK, GroEL or HspA, were affected. PMID- 11496004 TI - Starvation survival in Listeria monocytogenes: characterization of the response and the role of known and novel components. AB - The starvation survival response (SSR) of Listeria monocytogenes EGD is induced under glucose- or multiple-nutrient-, but not amino-acid limitation. 0.01-0.2% of the population remain viable even after 20 d and the survivors show a reduced cell size and increased cross-protection to several environmental stresses. The development of the SSR may therefore be important in L. monocytogenes survival in the food environment. The initiation, but not the maintenance, of the SSR involves both protein and cell wall biosynthesis. It is also likely that nutrients released from dead cells are recycled to allow survival of the remaining population. To define the molecular mechanisms involved in the initiation, maintenance and release from the SSR the role of known, and novel, components was examined. The well-characterized regulators SigB and PrfA are both required for the full SSR and effect stress resistance during growth and starvation. A transposon mutagenesis screen identified two novel loci with roles in the SSR and stress resistance. Characterization of the transposon insertion sites revealed a putative homologue of the gene yulB from Bacillus subtilis and a gene of unknown function. The potential individual and combined roles of the SSR components are discussed. PMID- 11496005 TI - Characterization of the low-pH responses of Helicobacter pylori using genomic DNA arrays. AB - Helicobacter pylori is unique among bacterial pathogens in its ability to persist in the acidic environment of the human stomach. To identify H. pylori genes responsive to low pH, the authors assembled a high-density array of PCR-amplified random genomic DNA. Hybridization of radiolabelled cDNA probes, prepared using total RNA from bacteria exposed to buffer at either pH 4.0 or pH 7.0, allowed both qualitative and quantitative information on differential gene expression to be obtained. A previously described low-pH-induced gene, cagA, was identified together with several novel genes that may have relevance to the survival and persistence of H. pylori in the gastric environment. These include genes encoding enzymes involved in LPS and phospholipid synthesis and secF, encoding a component of the protein export machinery. A hypothetical protein unique to H. pylori (HP0681) was also found to be acid induced. Genes down-regulated at pH 4.0 include those encoding a sugar nucleotide biosynthesis protein, a flagellar protein and an outer-membrane protein. Differential gene expression was confirmed by total RNA slot-blot hybridization. PMID- 11496006 TI - cDNA-RNA subtractive hybridization reveals increased expression of mycocerosic acid synthase in intracellular Mycobacterium bovis BCG. AB - Identifying genes that are differentially expressed by Mycobacterium bovis BCG after phagocytosis by macrophages will facilitate the understanding of the molecular mechanisms of host cell-intracellular pathogen interactions. To identify such genes a cDNA-total RNA subtractive hybridization strategy has been used that circumvents the problems both of limited availability of bacterial RNA from models of infection and the high rRNA backgrounds in total bacterial RNA. The subtraction products were used to screen a high-density gridded Mycobacterium tuberculosis genomic library. Sequence data were obtained from 19 differential clones, five of which contained overlapping sequences for the gene encoding mycocerosic acid synthase (mas). Mas is an enzyme involved in the synthesis of multi-methylated long-chain fatty acids that are part of phthiocerol dimycocerosate, a major component of the complex mycobacterial cell wall. Northern blotting and primer extension data confirmed up-regulation of mas in intracellular mycobacteria and also revealed a putative extended -10 promoter structure and a long untranslated upstream region 5' of the mas transcripts, containing predicted double-stranded structures. Furthermore, clones containing overlapping sequences for furB, groEL-2, rplE and fadD28 were identified and the up-regulation of these genes was confirmed by Northern blot analysis. The cDNA RNA subtractive hybridization enrichment and high density gridded library screening, combined with selective extraction of bacterial mRNA represents a valuable approach to the identification of genes expressed during intra macrophage residence for bacteria such as M. bovis BCG and the pathogenic mycobacterium, M. tuberculosis. PMID- 11496007 TI - Serine/threonine protein kinases PknF and PknG of Mycobacterium tuberculosis: characterization and localization. AB - Pathogenesis of Mycobacterium tuberculosis is closely connected to its survival and replication within the host. Some pathogenic bacteria employ protein kinases that interfere with the cellular signalling network of host cells and promote bacterial survival. In this study, the pknF and pknG genes, which encode two putative protein kinases of M. tuberculosis H(37)Rv, protein kinase F (PknF) and protein kinase G (PknG), respectively, were cloned and expressed in Escherichia coli. Purified PknF phosphorylated the peptide substrate myelin basic protein (MBP) at serine and threonine residues, while purified PknG phosphorylated only at serine residues. The activity of the two kinases was abrogated by mutation of the codon for the predicted ATP-binding-site lysine residue. Southern blot analysis revealed that homologues of the genes encoding the two kinases are present in M. tuberculosis H(37)Ra and Mycobacterium bovis BCG, but not in Mycobacterium smegmatis. Immunoblot analysis of various cellular fractions of M. tuberculosis H(37)Rv revealed that PknF is a transmembrane protein and that PknG is predominantly a cytosolic enzyme. The present study should aid in elucidating the role of these protein kinases in the pathogenesis of mycobacteria. PMID- 11496008 TI - Erp, an extracellular protein family specific to mycobacteria. AB - Erp (exported repeated protein) was originally characterized as a virulence factor in Mycobacterium tuberculosis and was thought to be present only in Mycobacterium leprae and members of the TB complex. Here it is shown that Erp is a ubiquitous extracellular protein found in all of the mycobacterial species tested. Erp proteins have a modular organization and contain three domains: a highly conserved amino-terminal domain which includes a signal sequence, a central variable region containing repeats based on the motif PGLTS, and a conserved carboxy-terminal domain rich in proline and alanine. The number and fidelity of PGLTS repeats of the central region differ considerably between mycobacterial species. This region is, however, identical in all of the clinical M. tuberculosis strains tested. In addition, it is shown here that a Mycobacterium smegmatis erp::aph mutant displays altered colony morphology which is complemented by all the Erp orthologues tested. The genome sequence flanking the erp gene includes cell-wall-related ORFs and displays extensive conservation between saprophytic and pathogenic mycobacteria. PMID- 11496009 TI - Comparative whole-genome analyses reveal over 100 putative phase-variable genes in the pathogenic Neisseria spp. AB - Previously, a complete genome analysis of Neisseria meningitidis strain MC58 revealed the largest repertoire of putative phase-variable genes described in any species to date. Initial comparisons with two incomplete Neisseria spp. genome sequences available at that time revealed differences in the repeats associated with these genes in the form of polymorphisms, the absence of the potentially unstable elements in some alleles, and in the repertoire of the genes that were present. Analyses of the complete genomes of N. meningitidis strain Z2491 and Neisseria gonorrhoeae strain FA1090 have been performed and are combined with a comprehensive comparative analysis between the three available complete genome sequences. This has increased the sensitivity of these searches and provided additional contextual information that facilitates the interpretation of the functional consequences of repeat instability. This analysis identified: (i) 68 phase-variable gene candidates in N. meningitidis strain Z2491, rather than the 27 previously reported; (ii) 83 candidates in N. gonorrhoeae strain FA1090; and (iii) 82 candidates in N. meningitidis strain MC58, including an additional 19 identified through cross-comparisons with the other two strains. In addition to the 18 members of the opa gene family, a repertoire of 119 putative phase variable genes is described, indicating a huge potential for diversification mediated by this mechanism of gene switching in these species that is central to their interactions with the host and environmental transitions. Eighty-two of these are either known (14) or strong (68) candidates for phase variation, which together with the opa genes make a total of 100 identified genes. The repertoires of the genes identified in this analysis diverge from the different species groupings, indicating horizontal exchange that significantly affects the species and strain complements of these genes. PMID- 11496010 TI - Inverse relationship between pilus-mediated gonococcal adherence and surface expression of the pilus receptor, CD46. AB - Pilus-mediated adherence to mucosal epithelial cells is a critical step for Neisseria gonorrhoeae to establish an infection in the human host. CD46, the defined receptor for the gonococcal pilus, is a complement-regulatory protein that is expressed on all human nucleated cells. It was observed that a piliated, Opa(-) variant of gonococcal strain FA1090 adhered with different efficiencies to the human epithelial cell lines tested (Chang, ME180, HEC-1B and PC-3). Surprisingly, these differences in adherence levels did not correlate with levels of CD46 expressed by these cell lines. In fact, there was an inverse relationship between total surface-exposed CD46 and gonococcal adherence. Four major isoforms of CD46 are produced due to alternative RNA splicing of a surface-exposed region and the cytoplasmic tail. The relative isoform surface expression of each cell line was determined, and each was found to express different ratios of the four CD46 isoforms. No correlation could be derived between CD46 isoform surface expression and pilus-mediated gonococcal adherence, indicating that CD46 does not act as a classic receptor for gonococcal pili. PMID- 11496011 TI - Association between intimin (eae) and EspB gene subtypes in attaching and effacing Escherichia coli strains isolated from diarrhoeic lambs and goat kids. AB - Attaching and effacing Escherichia coli (AEEC) strains isolated from diarrhoeic lambs and goat kids were characterized for intimin (eae) and EspB (espB) gene subtypes by PCR and sequencing, and for genetic relatedness by PFGE. Fifty (23 ovine and 27 caprine) AEEC strains of 398 (246 ovine and 152 caprine) analysed were detected by colony blot hybridization. These strains were epidemiologically unrelated since they were isolated from different outbreaks of neonatal diarrhoea over a long period. Ovine AEEC strains belonged to serogroups O2, O4, O26, O80, O91 or were untypable, and caprine strains belonged to serogroups O3, O153 and O163. Two intimin subtypes were detected among the ovine and caprine strains studied. Most of the strains (43/50) had the beta type intimin gene, but seven ovine strains possessed a variant gamma type intimin gene (gamma(V)). Analysis of deduced amino acid sequences of the eae gene revealed that the sequences of beta intimin of ovine and caprine strains were virtually identical to those of beta intimin of rabbit EPEC, human EPEC clone 2 and swine AEEC, whereas the gamma(V) intimin present in seven ovine strains had 75-76% identity with gamma intimin of human EHEC clone 1 strains, and 96% of identity with intimin of the human EHEC strain 95NR1 of serotype O111:H-. A PCR test was developed to identify the three different espB gene subtypes, espB of human EPEC clone 1 (espBalpha), espB of human EHEC clone 1 (espBgamma) and espB of rabbit EPEC and human EPEC clone 2 (espBbeta). There was close correlation between the intimin beta type and the espBbeta gene subtype in the ovine and caprine AEEC strains. The seven ovine strains possessing the gamma(V) intimin gene possessed the espBalpha gene subtype. None of the strains studied possessed the espBgamma gene found in human O157:H7 EHEC strains. PFGE analysis of genomic DNA of selected strains showed a great diversity among strains. Cluster analysis of PFGE patterns showed greater divergence between strains with the gamma(V) intimin gene than between strains with the beta intimin gene. This study showed that most of the AEEC strains isolated from diarrhoeic lambs and goat kids possessed beta intimin and espB genes identical to those of rabbit EPEC, and they may be associated with enteric disease in small ruminants. PMID- 11496012 TI - Dynamic changes in the morphology of Cryptococcus neoformans during murine pulmonary infection. AB - The pathogenesis of Cryptococcus neoformans infection has been studied extensively with respect to inflammatory and pathological changes, but very little information is available regarding the morphology of yeast cells during the course of infection. Electron microscopy of Cryptococcus neoformans in murine pulmonary infection revealed increased cell wall thickness with time, but this difference was only partially accounted for by increases in cell diameter. Cell walls of melanized cells were thicker than those of nonmelanized cells 2 h after infection, and the cell wall of yeast became blacker with time, suggesting that melanization contributes to the increased cell wall thickness. Heterogeneous cell populations emerged, with the appearance of giant forms. While for C. neoformans ATCC strain 24067 (serotype D) the full spectrum of cell sizes were observed, for strains H99 (serotype A) and 3501 (serotype D) cells were divisible into two populations, giant and micro forms. In contrast to cellular heterogeneity, the epitope recognized by a protective mAb on the capsular glucuronoxylomannan (GXM) was found at all times of infection. Immunoelectron microscopy using mAbs to GXM demonstrated reactivity with intracellular structures, suggesting that synthesis of capsular polysaccharide occurs, at least in part, in the cytoplasm. In summary, the results indicate that: (i) the infection is dynamic with respect to yeast cell morphology; (ii) giant cell forms arise in tissue during the course of infection; (iii) cell walls blacken and thicken during the course of infection, consistent with melanin synthesis during infection; and (iv) GXM epitopes are found in the capsule, cell wall and cytoplasm, consistent with intracellular polysaccharide synthesis. The results indicate that the population of C. neoformans cells in tissue is in a highly dynamic state, implying that the immune system must confront cells with varying characteristics during the course of infection. PMID- 11496013 TI - gmhX, a novel gene required for the incorporation of L-glycero-D-manno-heptose into lipooligosaccharide in Neisseria meningitidis. AB - Lipooligosaccharide (LOS) is a critical virulence factor of Neisseria meningitidis. A Tn916 insertion mutant, designated 469, was found to exhibit a markedly truncated LOS of 2.9 kDa when compared by Tricine/SDS-PAGE to the parental LOS (4.6 kDa). Electrospray mass spectrometry analysis of 469 LOS revealed that it consisted of the deep rough, heptose-deficient structure, Kdo(2) lipid A. Sequencing of chromosomal DNA flanking the Tn916 insertion in mutant 469 revealed that the transposon had inserted into an ORF predicted to encode a 187 aa protein with sequence homology to the histidinol-phosphate phosphatase domain of Escherichia coli HisB and to a family of genes of unknown function. The gene, designated gmhX, is part of a polycistronic operon (ice-2) containing two other genes, nlaB and orfC. nlaB encodes a lysophosphatidic-acid acyltransferase and orfC is predicted to encode a N-acetyltransferase. Specific polar and non-polar gmhX mutations in the parental strain, NMB, exhibited the truncated LOS structure of mutant 469, and repair of gmhX mutants by homologous recombination with the wild-type gmhX restored the LOS parental phenotype. GmhX mutants demonstrated increased sensitivity to polymyxin B. GmhX mutants and other Kdo(2)-lipid A mutants also demonstrated increased sensitivity to killing by normal human serum but were not as sensitive as inner-core mutants containing heptose. In the genomes of Helicobacter pylori and Synechocystis, gmhX homologues are associated with heptose biosynthesis genes; however, in N. meningitidis, gmhX was found in a location distinct from that of gmhA, rfaD, rfaE, aut and rfaC. GmhX is a novel enzyme required for the incorporation of L-glycero-D-manno-heptose into meningococcal LOS, and is a candidate for the 2-D-glycero-manno-heptose phosphatase of the heptose biosynthesis pathway. PMID- 11496014 TI - The evolution of bacterial LuxI and LuxR quorum sensing regulators. AB - Quorum sensing is a widespread form of bacterial communication in which individual cells produce and respond to specific N-acyl homoserine lactone signal metabolites. The different autoinducer synthases that generate these signals and the receptor/activator proteins that mediate the cell's response to them constitute evolutionarily conserved families of regulatory proteins known as the LuxI and LuxR families, respectively. We have performed a phylogenetic analysis of 76 individual LuxI and LuxR homologues present in diverse members of the Gram negative Proteobacteria. The results were consistent with an early origin for these regulators during the evolution of the Proteobacteria, with functional pairs of luxI and luxR genes possibly coevolving as regulatory cassettes. In many cases, specific LuxI and LuxR family members appeared to have been inherited horizontally. In particular, those species containing multiple LuxI and/or LuxR homologues usually appeared to have obtained each individual homologue or functional pair of homologues from an independent source. Because multiple homologues interact to form regulatory cascades, this finding suggests that hierarchical signalling pathways can potentially evolve by the sequential integration of pre-existing regulatory circuits acquired from diverse sources. PMID- 11496018 TI - Non-herbal nutritional supplements-the next wave: a comprehensive review of risks and benefits for the C-L psychiatrist. AB - The continuing popularity of complementary medicine has led to the frequent appearance of new products in the marketplace. Non-herbal supplements are now a popular choice for patients seeking relief from a variety of medical conditions. As with herbal medicines, there are concerns about the safety of these products in those with physical illness. Clearly, consultation-liaison psychiatrists will encounter patients using non-herbal products or inquiring about them. This article seeks to provide knowledge about the risks and benefits of non-herbal supplements that consultation-liaison psychiatrists are likely to encounter. PMID- 11496019 TI - Prevalence and risk of depression and anxiety-related disorders during the first three years after heart transplantation. AB - Although poor psychological adjustment to organ transplantation appears to be a major contributor to reduced quality of life and increased physical morbidity, the prevalence and risk factors for psychiatric disorder have not been considered beyond the first 12-18 months after transplantation. The authors enrolled a representative sample of 191 heart transplant recipients in a prospective examination of the prevalence, clinical characteristics, and risk factors for DSM III-R major depressive disorder (MDD), generalized anxiety disorder (GAD), associated adjustment disorders, and posttraumatic stress disorder related to transplant (PTSD-T) during the 3 years postsurgery. Survival analysis indicates that cumulative risks for disorder onset were MDD, 25.5%; adjustment disorders, 20.8% (17.7% with anxious mood); PTSD-T, 17.0%; and any assessed disorder, 38.3%. There was only one case of GAD. PTSD-T onset was limited almost exclusively to the first year posttransplant. Episodes of MDD (but not anxiety disorders) that occurred later posttransplant (8 to 36 months postsurgery) were more likely than early posttransplant episodes to be treated with psychotropic medications. For both MDD and anxiety disorders, later episodes were less likely to be precipitated by transplant-related stressors than other life stressors. Factors increasing cumulative risk for psychiatric disorder posttransplant included pretransplant psychiatric history, female gender, longer hospitalization, more impaired physical functional status, and lower social supports from caregiver and family in the perioperative period. Risk factors' effects were additive; the presence of an increasing number of risk factors bore a dose-response relationship to cumulative risk of disorder. PMID- 11496020 TI - Fluoxetine in pathologic skin-picking: open-label and double-blind results. AB - Various studies suggest that selective serotonin reuptake inhibitors (SSRIs) may be useful in treating pathologic skin-picking. The authors investigated the effectiveness of fluoxetine in treating this behavior. Fifteen subjects with clinically significant skin-picking were recruited by newspaper advertisement. They received 6 weeks of open-label treatment with fluoxetine. Responders were then randomized to 6 weeks of double-blind fluoxetine or placebo. Treatment effect was assessed with standardized rating scales. All 15 subjects completed open-label treatment, and 8 were responders. Of these eight, the four randomized to double-blind fluoxetine maintained clinically significant improvement. The four randomized to placebo returned to their baseline symptom level. Larger studies are needed to determine which individuals are likely to respond to fluoxetine and the relative effectiveness of fluoxetine, other SSRIs, and other forms of treatment. PMID- 11496021 TI - Personality dysfunction among somatizing patients. AB - To examine the nature and extent of personality dysfunction related to somatization, the authors administered the Structured Interview for DSM-IV Personality and the NEO Five-Factor Inventory to a series of somatizing and nonsomatizing patients in a general medicine clinic. A greater percentage of somatizers met criteria for one or more DSM-IV personality disorders, especially obsessive-compulsive disorder, than did control patients. Somatizers also differed from control patients with respect to self-defeating, depressive, and negativistic personality traits and scored higher on the dimension of neuroticism and lower on the dimension of agreeableness. In addition, initial and facultative somatizers showed more personality pathology than true somatizers. These findings suggest that certain personality disorders and traits contribute to somatization by way of increased symptom reporting and care-seeking behavior. PMID- 11496022 TI - A psychological profile of depressed and nondepressed women at high risk for breast cancer. AB - This study examines the difference on several demographic and psychosocial variables between women at high risk for breast cancer above and below the cut off point of a depression measure (Center for Epidemiological Study Depression Scale). Data are presented for 430 consecutive patients from the UCLA Revlon Breast Center High Risk Clinic. Women scoring above the depression cut-off point were younger, had more relatives with breast cancer, reported more symptoms of anxiety, and had more self-perceived vulnerability to breast cancer. In addition, women above the depression cut-off point were more likely to be single, childless, to have not viewed the results of the surgical treatment of their relative, and to feel more anxiety regarding screening practices (mammography, pap smears, and breast self-examinations). PMID- 11496023 TI - Psychiatric disorders before and after living-related transplantation. AB - The authors examined psychiatric disorders among two samples of patients who underwent living-related transplant (LRT) for kidney and liver failure. The postoperative prevalence of psychiatric disorders for adult transplant recipients was highest the first 3 months posttransplant. The incidence of psychiatric disorders in the adult recipients with living-related liver transplant (LRLT) was higher (54%, 22 of 41) than that of adult recipients with living-related kidney transplant (LRKT) (28%, 65 of 234). Twelve (80%) of the 15 adult LRLT recipients with adult child-to-parent donors exhibited paradoxical psychiatric syndrome (PPS). Among the 12 affected recipients, guilt-based psychiatric disorders of various types occurred despite successful operative outcome for both donor and recipient. The higher rate of psychiatric disorders among adult LRLT recipients was associated with the occurrence of PPS among recipients of an adult-child allograft. These results signal a new challenge for consultation psychiatrists working with transplant patients. PMID- 11496024 TI - Nocturnal pruritus in a cardiac pretransplant patient. PMID- 11496025 TI - Use of quetiapine in delirium: case reports. PMID- 11496026 TI - A case report of Kleine-Levin syndrome in an adolescent girl. PMID- 11496027 TI - Treatment contracts for patients with hepatitis C, psychiatric illness, and substance abuse. PMID- 11496028 TI - Pseudocataplexy. PMID- 11496029 TI - Lithium-induced hypercalcemia and parathyroid dysfunction. PMID- 11496030 TI - Electroconvulsive therapy for depression in a cardiac transplant patient. PMID- 11496031 TI - Suicidal ideation with IFN-alpha and ribavirin in a patient with hepatitis C. PMID- 11496032 TI - Acute neutropenia in a patient treated with quetiapine. PMID- 11496033 TI - Digitalis intoxication misdiagnosed as depression--revisited. PMID- 11496034 TI - Lack of citalopram effect on the pharmacokinetics of cyclosporine. PMID- 11496035 TI - Olanzapine-lithium encephalopathy. PMID- 11496036 TI - Medicare, the Internet, and the future of telemedicine. AB - OBJECTIVES: Beginning October 1, 2001, the Health Care Financing Administration (HCFA) will extend Medicare coverage to a wide range of telemedicine services and providers, allowing for medical visits, consultations, mental health services, and pharmacologic monitoring of patients living in rural areas. Payment to providers will be at a rate similar to that paid without the use of telemedicine. Furthermore, Medicare will pay a facility fee of $20 to the originating site per telemedicine session. This article discusses how advances in computer connectivity and communication infrastructure, coupled with Medicare reimbursement for telemedicine services, present medical providers with a unique opportunity to improve healthcare delivery to their patients, in particular those living in rural counties in the United States. DATA SOURCES: Peer-reviewed articles published in the literature and industry-specific surveys published on the Internet. CONCLUSIONS: There is little doubt that recent changes in HCFA reimbursement for telemedicine will have a dramatic impact on the delivery of medical care to rural America. By correcting the mistakes of the 1997 Balanced Budget Act provisions, Congress has acknowledged telemedicine as a viable, potentially life-saving technology. The most likely scenario for the expansion of telemedicine services to rural counties will be through networks using Internet technology. The expansion of the Internet and broadband infrastructure should allow for the establishment of geographically wide and technically robust telemedicine networks, with a minimum of expense. PMID- 11496037 TI - Trends in technology and the future intensive care unit. AB - As advanced as today's critical care units are, changes in networks, computing platforms, instrumentation, human-machine interfaces, and software infrastructures and work being done in interoperability, process automation, and decision support all promise to alter both the structure and the operation of future critical unit settings. Although, as Yogi Berra said, "It's tough to make predictions, especially about the future," we can gain of a sense of the possibilities by studying emerging trends in each of these information technology related areas. After describing notional physical and functional architectures for today's critical care units, this article provides a high-level overview of a number of trends in the areas listed above and provides the reader with starting points for further study. Although each of these trends may be fairly unremarkable when taken in isolation, when considered as a whole, the implications for critical care are profound. To that end, a final section of the article speculates on specific ways that these trends are likely to play themselves out in the context of tomorrow's critical care environments. PMID- 11496038 TI - Telecardiology: potential impact on acute care. AB - Echocardiography is often used to diagnose and exclude important cardiac diagnoses in adults and children. Evolving telemedicine technology has the potential to improve access to echocardiography diagnoses in the intensive care unit, emergency room, and newborn nursery. The two primary modes of telemedicine practice are "store and forward" and "real-time" videoconferencing. A digital echocardiogram (often several one cardiac cycle loops) can be stored at one site and forwarded across a telemedicine network to a receiving station for review at a later time. Pediatric cardiologists often favor "real-time" telemedicine because of the ability to guide sonographers with limited experience in congenital heart disease. A complete telemedicine system requires a modified computer, a low- or high-speed connection, and telemedicine inputs. Several adult and pediatric clinical studies have shown telemedicine to be accurate and cost effective, improve patient care, enhance echocardiogram quality and sonographer proficiency, and promote practice expansion. Obstacles to widespread implementation of telemedicine include lack of standardization of telemedicine components, confusing legal issues and licensure requirements, and poor reimbursement. PMID- 11496039 TI - The Internet, the electronic medical record, the pediatric intensive care unit, and everything. AB - This article details how computers have changed life for those of us in pediatric intensive care. A week of clinical activity is described, with a focus on the interactions with computer systems that have become an integral part of patient care activities for many of us. It becomes clear that the boundaries between personal computers, hospital systems, and the Internet are often not sharply defined. Resources that are used every week may include those residing on a personal digital assistant, on the hospital's electronic medical record, or on a distant site on the World Wide Web. Key resources on the Internet (World Wide Web and e-mail) are identified. The technical underpinnings, particularly the network that provides the infrastructure for various resources, are described. PMID- 11496040 TI - Computer-assisted learning in critical care: from ENIAC to HAL. AB - Computers are commonly used to serve many functions in today's modern intensive care unit. One of the most intriguing and perhaps most challenging applications of computers has been to attempt to improve medical education. With the introduction of the first computer, medical educators began looking for ways to incorporate their use into the modern curriculum. Prior limitations of cost and complexity of computers have consistently decreased since their introduction, making it increasingly feasible to incorporate computers into medical education. Simultaneously, the capabilities and capacities of computers have increased. Combining the computer with other modern digital technology has allowed the development of more intricate and realistic educational tools. The purpose of this article is to briefly describe the history and use of computers in medical education with special reference to critical care medicine. In addition, we will examine the role of computers in teaching and learning and discuss the types of interaction between the computer user and the computer. PMID- 11496041 TI - The eICU: it's not just telemedicine. AB - Intensive care units (ICUs) are major sites for medical errors and adverse events. Suboptimal outcomes reflect a widespread failure to implement care delivery systems that successfully address the complexity of modern ICUs. Whereas other industries have used information technologies to fundamentally improve operating efficiency and enhance safety, medicine has been slow to implement such strategies. Most ICUs do not even track performance; fewer still have the capability to examine clinical data and use this information to guide quality improvement initiatives. This article describes a technology-enabled care model (electronic ICU, or eICU) that represents a new paradigm for delivery of critical care services. A major component of the model is the use of telemedicine to leverage clinical expertise and facilitate a round-the-clock proactive care by intensivist-led teams of ICU caregivers. Novel data presentation formats, computerized decision support, and smart alarms are used to enhance efficiency, increase effectiveness, and standardize clinical and operating processes. In addition, the technology infrastructure facilitates performance improvement by providing an automated means to measure outcomes, track performance, and monitor resource utilization. The program is designed to support the multidisciplinary intensivist-led team model and incorporates comprehensive ICU re-engineering efforts to change practice behavior. Although this model can transform ICUs into centers of excellence, success will hinge on hospitals accepting the underlying value proposition and physicians being willing to change established practices. PMID- 11496042 TI - Measuring the impact of new technology: an outcomes-based approach. AB - In the current era of decreasing resources, many healthcare organizations are under increasing pressure to evaluate new technology. Advances and innovations in healthcare technology are instrumental in reshaping the healthcare system and in impacting the practice of medicine, including critical care. Healthcare organizations are faced with the following four questions related to adopting a new technology: 1) Should the organization invest in this technology? 2) What are the associated benefits and risks of the capital investment? 3) What is the impact on patient outcomes as a result of adopting the technology? 4) What is the return on investment to the organization? These issues represent challenging areas for the researcher, clinician, manager, and policy maker. New techniques from the business and outcomes research arenas allow for the development of quantitative and qualitative models to facilitate answering these questions. By integrating business and clinical considerations, these models can impact policy at two levels: 1) by identifying optimal capital expenditure decisions for individual healthcare organizations, it can provide a significant competitive advantage for an organization; and 2) by identifying optimal health policies related to technology at a national and international health level, it can impact the healthcare systems and patient outcomes in a favorable manner. PMID- 11496043 TI - Online personal medical records: are they reliable for acute/critical care? AB - OBJECTIVE: To provide an introduction to Internet-based Online Personal Medical Records (OPMRs), to assess their use and limitations in acute/critical care situations, and to identify potential improvements that could increase their usefulness. DESIGN: A review of publicly available Internet-based OPMRs conducted in April 2001. DATA SOURCES: Twenty-nine OPMR sites were identified in March 2000 using ten Internet search engines with the search term "Personal Medical Records." Through 2000 and 2001, an additional 37 sites were identified using lists obtained from trade journals and through the author's participation in standards-setting meetings. MEASUREMENTS: Each publicly available site was reviewed to assess suitability for acute/critical care situations using four measures developed by the author and for general use using eight measures developed in a standards-setting process described in the article. RESULTS: Of the 66 companies identified, only 16 still offer OPMRs that are available to the public on the Internet. None of these met all of the evaluation measures. Only 19% had rapid emergency access capabilities and only 63% provided medical summaries of the record. Security and confidentiality issues were well addressed in 94% of sites. Data portability was virtually nonexistent because all OPMRs lacked the ability to exchange data electronically with other OPMRs, and only two OPMRs permitted data transfer from physician electronic medical records. Controls over data accuracy were poor: 81% of sites allowed entry of dates for medical treatment before the patient's date of birth, and one site actually gave incorrect medical advice. OPMRs were periodically inaccessible because of programming deficiencies. Finally, approximately 40 sites ceased providing OPMRs in the past year, with the probable loss of patient information. CONCLUSIONS: Most OPMRs are not ready for use in acute/critical care situations. Many are just electronic versions of the paper-based health record notebooks that patients have used for years. They have, however, great promise and, with further development, could form the basis of a new medical record system that could contribute to improving the quality of medical care. PMID- 11496045 TI - Serum cystatin C as a marker of glomerular filtration rate. PMID- 11496046 TI - Molecular aspects in the regulation of renal inorganic phosphate reabsorption: the type IIa sodium/inorganic phosphate co-transporter as the key player. AB - The type IIa sodium/inorganic phosphate co-transporter is the rate-limiting inorganic phosphate transport pathway in renal brush-border membranes, and is thus a key player in overall inorganic phosphate homeostasis. Its regulation is mostly associated with membrane retrieval/reinsertion (traffic) of the transport protein. This membrane traffic is controlled by specific 'motifs' at the level of the transporter protein and probably involves interacting proteins (e.g. for scaffolding, regulation or sorting). The intracellular signaling mechanisms (e.g. the involvement of kinases) and the involvement of the cytoskeleton are not yet understood. Hereditary alterations in renal inorganic phosphate handling can be associated with factors controlling the expression of the brush-border type IIa sodium/inorganic phosphate co-transporter. PMID- 11496047 TI - Consequences of hyperphosphatemia and elevated levels of the calcium-phosphorus product in dialysis patients. AB - Control of serum phosphorus levels is a central goal in the management of patients with chronic renal failure. Inadequate control of serum phosphorus leads to elevated levels of the calcium-phosphorus product. This plays a pivotal role in vascular calcification, cardiovascular disease, calciphylaxis, and death. Elevated phosphorus and elevated levels of the calcium-phosphorus product are both significant predictors of cardiovascular mortality, at phosphorus and calcium-phosphorus product levels that were considered safe until recently. A lowering of levels such that phosphorus is maintained between 2.2 and 5.5 mg/dl, calcium-phosphorus product is below 55 mg(2)/dl(2), and serum calcium is at 9.2 9.6 mg/dl, respectively, might well be the goal of therapeutic management strategies. PMID- 11496048 TI - The evolution of assays for parathyroid hormone. AB - Recent progress in the assay of parathyroid hormone has revealed that commercially available assays for intact parathyroid hormone also measure additional parathyroid hormone peptides that appear to be truncated at the amino terminal region and have the elution position on high-performance liquid chromatography of parathyroid hormone 7-84. Specific assays have been developed that measure only the true or 'whole', 84-amino-acid peptide. Such 'whole' parathyroid hormone assays have led to the discovery of new findings that suggest that parathyroid hormone fragments such as parathyroid hormone 7-84, which have hitherto been considered to be biologically inactive, may actually have biologic effects. These data, coupled with the emerging discovery of additional receptors for parathyroid hormone peptides, suggest that parathyroid hormone fragments might have potentially important actions, at least in the setting of renal failure. PMID- 11496049 TI - Calcimimetic agents for the treatment of hyperparathyroidism. AB - Calcimimetic agents are small organic molecules that act as allosteric activators of the calcium sensing receptor. They lower the threshold for receptor activation by extracellular calcium ions and, in parathyroid cells, diminish parathyroid hormone secretion. Calcimimetic compounds represent a novel class of therapeutic agents that may provide a way of controlling excess parathyroid hormone secretion in several clinical disorders. Although experience from clinical trials in humans is limited, available data suggest that calcimimetic agents effectively lower plasma parathyroid hormone levels in patients with primary hyperparathyroidism and those with secondary hyperparathyroidism caused by end-stage renal disease. Calcimimetic compounds thus have considerable potential as a new approach to the medical management of several important clinical disorders of bone and mineral metabolism. PMID- 11496050 TI - Bisphosphonates in renal osteodystrophy. AB - The present review considers the role that bisphosphonates might have in patients with renal failure. Although bisphosphonates are widely used to reduce fracture risk in patients with osteoporosis, few studies have documented their effect in patients with renal osteodystrophy. The pathogenesis of bone loss after renal transplantation and the role of the recently identified osteoprotegerin/receptor activating nuclear factor-kappaB system is described. Inhibition of bone resorption may prove beneficial when high bone turnover is present, but there are potential drawbacks to widespread use of bisphosphonates. These issues are discussed, with emphasis placed on reports published within the past 18 months. PMID- 11496051 TI - Glucocorticoid-induced osteoporosis. AB - Glucocorticoid-induced osteoporosis is a significant problem in patients receiving glucocorticoids after transplantation and for the treatment of parenchymal renal disease and rheumatological disorders. Frequently, patients are not evaluated or treated appropriately for glucocorticoid-induced osteoporosis. Bone loss occurs early after the administration of high-dose glucocorticoid therapy. Elderly patients with low bone densities before the initiation of glucocorticoid therapy are at particular risk of developing significant bone loss that could result in fractures. New information is now available concerning the mechanisms by which glucocorticoid-associated bone disease occurs. New therapies with anti-resorptive agents such as bisphosphonates and with anabolic agents such as parathyroid hormone offer the prospect of effective treatment of glucocorticoid-induced osteoporosis. PMID- 11496052 TI - The Heart Outcomes Prevention Evaluation study: angiotensin-converting enzyme inhibitors: are their benefits a class effect or do individual agents differ? AB - The Heart Outcomes Prevention Evaluation study was a landmark study employing the angiotensin-converting enzyme inhibitor ramipril in a patient population pre destined for vascular events. This study found that a 10 mg dose of ramipril in comparison with placebo reduced the incidence of death, myocardial infarction, stroke, and death from cardiovascular causes by 22%. This improvement in outcome was viewed as a direct consequence of ramipril, although the fact that blood pressure was reduced with ramipril has cast some considerable doubt on this supposition. Whether the observed findings in this study are a 'class effect' for all angiotensin-converting enzyme inhibitors is unclear. To its credit, ramipril is the first angiotensin-converting enzyme inhibitor shown to prevent ischemic events in high-risk patients without discernible left ventricular dysfunction. Other similar trials are underway, which are studying similar populations to those included in this landmark study and should resolve the question of whether the cardioprotective effects of angiotensin-converting enzyme inhibitors are a 'class effect'. PMID- 11496053 TI - Renal effects of cyclooxygenase-2 selective inhibitors. AB - Recent in-vitro and animal data show that cyclooxygenase-2 has an integral role in the physiology and pathophysiology of the kidney. Cyclooxygenase-2 regulates renin-angiotensin secretion, and thereby glomerular filtration rate and sodium homeostasis. It is also important for protecting against hypertonic stress. As a consequence, it is not surprising that clinical data verify that selective inhibitors of cyclooxygenase-2 affect renal function to a degree similar to that which has previously been documented with nonselective nonsteroidal anti inflammatory drugs. PMID- 11496054 TI - Nocturnal non-dipping: what does it augur? AB - The non-dipping pattern of blood pressure (defined as a nocturnal fall of less than 10%) occurs in about 25% of hypertensives, with increased prevalence in certain sub-groups such as diabetics, African-Americans, and patients with renal disease. It almost certainly has multiple causes, including factors such as the levels of activity and arousal during both the day and the night, the depth and quality of sleep, and the activity of the sympathetic nervous system, among others. In patients with uncomplicated hypertension, the reproducibility is relatively low. There is evidence suggesting that the non-dipping pattern may have an adverse prognosis: thus, it appears to predict the progression of renal disease, to be associated with increased target-organ damage (in some studies), and also to predict increased cardiovascular morbidity. Antihypertensive drug treatment can normalize the non-dipping pattern, but the therapeutic consequences of this are unknown. PMID- 11496055 TI - Effects of antihypertensive drugs on vascular remodeling: do they predict outcome in response to antihypertensive therapy? AB - Remodeling of large and small arteries in hypertension contributes to elevation of blood pressure, and may participate in the complications of hypertension. Large arteries exhibit increased lumen size, thickened media with increased collagen deposition, and decreased compliance, which contributes to raised systolic blood pressure and pulse pressure. In small (resistance) arteries smooth muscle cells are restructured around a smaller lumen, without true hypertrophy, particularly in milder forms of hypertension, whereas in severe forms and in secondary hypertension hypertrophic remodeling has been reported. Endothelial dysfunction occurs in many patients, with prevalence similar to that of left ventricular hypertrophy. Treatment with angiotensin-converting enzyme inhibitors, angiotensin receptor subtype 1 antagonists and long-acting calcium channel blockers has corrected changes in large and small arteries in hypertensive patients. Treatment with beta-blockers did not modify either structure or function of small arteries. Improved outcomes were reported in clinical trials with drugs that exert vascular protective effects, such as angiotensin-converting enzyme inhibitors and angiotensin receptor subtype 1 antagonists, as well as with those that do not appear to improve vascular structure or function. Recent trials suggest that these different drugs may provide similar benefits essentially through blood pressure lowering, although some minor differences between drugs have been noted. For example, the alpha-blocker doxasozin has been associated with worse outcomes (heart failure) than have diuretics. That hard end-point clinical trials have not demonstrated any advantages of agents with vasculoprotective properties may relate in part to the relatively short duration of some of these multicenter trials (3-5 years). Another contributing factor may be the low number of events with each drug class in the longer trials. Thus, current evidence does not support the rational expectation that vasculoprotective antihypertensive agents will be associated with better outcomes in hypertensive patients, possibly because of limitations of these trials. PMID- 11496056 TI - Does aspirin attenuate the effect of angiotensin-converting enzyme inhibitors in hypertension or heart failure? AB - There is a wealth of data that suggests an important interaction between aspirin and angiotensin-converting enzyme inhibitors in patients with chronic stable cardiovascular disease. The interaction is less obvious in the postinfarction setting, possibly reflecting the fact that many patients stop their aspirin therapy within a few months of such an event. An interaction is biologically plausible, because there is considerable evidence that angiotensin-converting enzyme inhibitors exert important effects through increasing the production of vasodilator prostaglandins, whereas aspirin blocks their production through inhibition of cyclooxygenase, even at low doses. There is some evidence that low dose aspirin may raise systolic and diastolic blood pressure. There is also considerable evidence that aspirin may entirely neutralize the clinical benefits of angiotensin-converting enzyme inhibitors in patients with heart failure. In addition, aspirin may have an adverse effect on outcome in patients with heart failure that is independent of any interaction with angiotensin-converting enzyme inhibitors, possibly by blocking endogenous vasodilator prostaglandin production and enhancing the vasoconstrictor potential of endothelin. The evidence is not sufficient to justify advising long-term aspirin therapy for patients with cardiovascular disease in general, and for those with heart failure in particular. Thus, the lack of evidence of benefit with aspirin in patients with heart failure and coronary disease, along with growing evidence that aspirin is directly harmful in patients with heart failure and that aspirin may negate the benefits of angiotensin-converting enzyme inhibitors suggest that, unless there is an opportunity to randomize the patient into a study of antithrombotic strategies, then aspirin should be withdrawn or possibly substituted with an anticoagulant or an antiplatelet agent that does not block cyclooxygenase. In contrast, there is fairly robust evidence for a benefit of both aspirin and angiotensin-converting enzyme inhibitors during the first 5 weeks after a myocardial infarction, with little evidence of an interaction. The combination of aspirin and angiotensin-converting enzyme inhibitors is warranted during this period, after which discontinuation or substitution of aspirin with another agent should be considered. PMID- 11496057 TI - Cardiovascular effects of erythropoietin and anemia correction. AB - Although recombinant erythropoietin has no short-acting pressor effect in vivo, its long-term administration frequently raises arterial pressure in humans and animals, with renal insufficiency. Contrary to the original view, erythropoietin induced hypertension is not due to amelioration of anemia, because a similar rise in blood pressure occurs, despite persistent anemia, in erythropoietin-treated iron-deficient animals and humans. Moreover, multiple small blood transfusions administered to simulate the action of erythropoietin fail to increase blood pressure. Finally, iron repletion in severely anemic iron-deficient patients maintained on constant erythropoietin dosages does not raise blood pressure, despite a dramatic increase in hematocrit. Thus, chronic erythropoietin administration results in a hematocrit-independent, vasoconstriction-dependent hypertension that is marked by, and largely due to, elevated resting and agonist stimulated cytoplasmic calcium concentration, leading to resistance to the vasodilatory action of nitric oxide. In addition, increased endothelin production, upregulation of tissue (but not circulating) renin and angiotensinogen expression, and a possible change in vascular tissue prostaglandin production have been variably demonstrated with erythropoietin administration in humans, intact animals and cultured endothelial cells. Erythropoietin has been shown to promote angiogenesis and stimulate endothelial and vascular smooth muscle cell proliferation. Finally, partial correction of anemia with erythropoietin therapy may partly prevent or reverse left ventricular hypertrophy in dialysis-dependent and dialysis-independent patients with chronic renal insufficiency. However, data on the risks and benefits of complete correction of anemia in this population are limited and inconclusive, and await future investigation. PMID- 11496058 TI - n-3 Fatty acids and their role in nephrologic practice. AB - During the past year, a newly reported clinical trial has strengthened the argument for recommending daily treatment with n-3 polyunsaturated fatty acids in patients with immunoglobulin A nephropathy (the most common form of primary glomerulonephritis in the world) who are at high risk for progression of renal disease. Studies are underway that involve a combination of cyclosporine A, a commonly prescribed immunosuppressive agent in solid-organ transplantation, with a high-potency n-3 polyunsaturated fatty acid to reduce cyclosporine toxicity. Two studies reported during the past year show promise that dietary supplementation with n-3 polyunsaturated fatty acids will substantially decrease vascular access graft thrombosis in patients receiving maintenance hemodialysis, and may reduce hypercalciuria in patients who suffer from kidney stones. PMID- 11496059 TI - Is there a place for combining angiotensin-converting enzyme inhibitors and angiotensin-receptor antagonists in the treatment of hypertension, renal disease or congestive heart failure? AB - Angiotensin-converting enzyme inhibitors and angiotensin II receptor subtype 1 antagonists have proven to be effective and well tolerated antihypertensive agents. They also exhibit unique cardioprotective and renoprotective properties in patients with comorbid conditions such as congestive heart failure and proteinuria or renal insufficiency. This benefit is observed most dramatically in diabetic persons. Although inconclusive, the results of a limited number of clinical trials support the notion that additive antihypertensive, cardioprotective, and renoprotective effects may be obtained with combined used of angiotensin-converting enzyme inhibitors and angiotensin II receptor subtype 1 antagonists in some patients. More studies are needed to confirm the findings of these preliminary studies, and to define more clearly those subsets of patients who might derive the greatest benefit from angiotensin-converting enzyme inhibitor-angiotensin II receptor subtype 1 antagonist combination therapy. PMID- 11496060 TI - Evolving concepts in epithelial magnesium transport. AB - Magnesium is an important, predominantly intracellular cation that is required for a wide variety of cellular processes. The mammalian kidney plays a key role in whole-body magnesium homeostasis, but the molecular and cellular mechanisms that underlie renal epithelial magnesium reabsorption are poorly understood. Traditional physiologic approaches have been severely hampered by the lack of a useful radioisotope of magnesium that can be used for tracer flux studies. The present review discusses physiologic insights gained from recent reverse-genetic studies that have identified a plethora of genes involved in inherited renal magnesium wasting syndromes. PMID- 11496061 TI - Regulation of the sodium transporters NHE3, NKCC2 and NCC in the kidney. AB - The regulation of sodium transport in the kidney is important for maintenance of extracellular fluid volume and arterial blood-pressure regulation. The major sodium transporters and channels in individual renal tubule segments have been identified via physiological techniques, and complementary DNAs for all of the key sodium transporters and channels expressed along the renal tubule have been cloned. Complementary DNA probes and antibodies are now being used to investigate the molecular basis of renal tubule sodium-transport regulation. This review summarizes some of the major observations made in the past year that are relevant to the regulation of the major sodium transporters in the proximal tubule (the type 3 sodium-hydrogen exchanger, NHE3), the thick ascending limb of Henle (the bumetanide-sensitive sodium-potassium-chloride cotransporter, NKCC2), and the distal convoluted tubule (the thiazide-sensitive sodium-chloride cotransporter, NCC). PMID- 11496062 TI - A mineralocorticoid receptor mutation causing human hypertension. AB - In recent years, the study of monogenic forms of hypertension has added greatly to our understanding of the regulatory mechanisms affecting blood pressure. Recently, a novel such form of human hypertension caused by gain-of-function mutation in the mineralocorticoid receptor, the mediator of aldosterone-induced sodium transport in the distal nephron, has been described, with the notable finding being that pregnancy causes a severe worsening of blood pressure. In this review, the mechanism by which the mutation causes hypertension, and the implications these findings have for improved understanding of cardiovascular physiology and mineralocorticoid receptor biology, are discussed. PMID- 11496063 TI - Mediators of aldosterone action in the renal tubule. AB - The aldosterone-sensitive distal nephron extends from the second part of the distal convoluted tubule to the inner medullary collecting duct. As recently shown, aldosterone increases within two hours the abundance of the alpha-subunit of the epithelial sodium channel along the entire aldosterone-sensitive distal nephron, whereas it induces only in an initial portion of the aldosterone sensitive distal nephron an apical translocation of all three epithelial sodium channel subunits. This suggests that another factor or factors determines the length of the aldosterone-sensitive distal nephron portion in which aldosterone controls epithelial sodium channel surface expression. Since the glucocorticoid induced kinase SGK1 was identified as aldosterone-induced protein in 1999, it has been postulated to play a key regulatory role. The in-vivo localization of its induction to segment-specific cells of the aldosterone-sensitive distal nephron, and the in-vitro correlation of the amount of its hyperphosphorylated form with transepithelial sodium transport, support this hypothesis. Other recent studies unravel pathways other than those activated by aldosterone and insulin that impact on SGK1 expression and/or function, and thus shed some light onto the complex network that appears to control sodium transport. In view of the ongoing research, the question of how, and formally also whether, SGK1 acts on the epithelial sodium channel should be resolved in the near future. PMID- 11496064 TI - Molecular physiology of the renal chloride-formate exchanger. AB - Renal apical chloride-base exchangers are essential to electrolyte and acid-base homeostasis. Different functional isoforms of apical anion exchangers have been identified in kidney proximal tubule and cortical collecting duct. Included amongst these are the following: chloride-formate, chloride-oxalate, and chloride hydroxyl exchangers in proximal tubule; and chloride-bicarbonate exchanger in cortical collecting duct. Chloride-formate exchange, which was first identified in kidney proximal tubule, works in parallel with the apical sodium-hydrogen exchanger, and is thought to reabsorb the bulk of luminal chloride. Despite numerous studies, the molecular identities of apical chloride-base exchangers have remained unknown. Recent studies have identified a new class of anion exchangers, including pendrin (encoded by the PDS gene) and downregulated in adenoma (DRA, encoded by the DRA gene). Pendrin is expressed in the kidney, whereas DRA is not. Functional studies indicate that pendrin can function in chloride-formate and chloride-base exchange modes. It is unlikely that pendrin is the apical chloride-formate exchanger in the kidney proximal tubule. However, it is the only molecule that has been shown to mediate chloride-formate exchange. In the present review, recent studies regarding the renal distribution and membrane localization of pendrin, and its functional properties, including its roles in chloride reabsorption and base excretion, are addressed. PMID- 11496065 TI - Renal potassium-chloride cotransporters. AB - The electroneutral cotransport of potassium and chloride is mediated by potassium chloride transporters, which are encoded by members of the gene family of cation chloride cotransporters. A significant body of evidence argues for swelling activated, basolateral potassium-chloride transport in the proximal tubule and thick ascending limb, with a potential role in transepithelial salt transport. However, the lack of specific inhibitors has impeded progress in this area. The cloning of the four potassium-chloride cotransporter genes has sparked new interest in this transport pathway, and promises to yield novel insights into their roles in cellular and renal physiology. PMID- 11496066 TI - Regulation of ROMK trafficking and channel activity. AB - ROMK potassium channels are present in the thick ascending limbs and cortical collecting ducts of the kidney and are responsible for potassium ion efflux in these segments. ROMK channels are regulated by multiple signalling pathways. This review focuses on recent advances in the regulation of ROMK channels by phosphatidylinositol-4,5-bisphosphate and by membrane trafficking. Relevant publications on the related inward rectifier potassium channels will also be discussed. PMID- 11496067 TI - Consensus statement of definitions for anorectal physiology and rectal cancer: report of the Tripartite Consensus Conference on Definitions for Anorectal Physiology and Rectal Cancer, Washington, D.C., May 1, 1999. PMID- 11496068 TI - Prevalence and severity of urinary incontinence and pelvic genital prolapse in females with anal incontinence or rectal prolapse. AB - PURPOSE: The aim of this study was to determine the prevalence, severity, and associations between urinary incontinence and genital prolapse in females after surgery for fecal incontinence or rectal prolapse. METHODS: All patients who underwent surgery for fecal incontinence (Group I) or rectal prolapse (Group II) were compared with a control group of females (Group III) by 43 questions regarding demographic data, past medical and surgical history, and diagnosis and treatment of anal and urinary incontinence and genital and rectal prolapse. The type (stress, urge, and total) of urinary incontinence was determined and graded using an incontinence severity questionnaire (Individual Incontinence Impact Questionnaire). RESULTS: Overall response rate in the three groups of patients was 40.1 percent. The questionnaire was sent to 240 patients operated on for fecal incontinence or rectal prolapse, and 83 of them responded (34.5 percent). The patients were distributed into three groups: Group I consisted of 51 patients (mean age 56.7 +/- 14); Group II consisted of 32 patients (69.7 +/- 11); and Group III consisted of 40 patients (60.5 +/- 16). The prevalence of urinary incontinence in Group I was 27 (54 percent), in Group II was 21 (65.6 percent), and Group III was 12 patients (30 percent; P = 0.003). Genital prolapse was present in 9 (17.6 percent), 11 (34.3 percent), and 5 patients (12.5 percent), respectively (P = 0.03). The prevalence of coexistent urinary incontinence and genital prolapse in both study groups was 22.8 percent (19 patients). There were no statistically significant differences between Groups I and II relative to prevalence, type, and severity of urinary incontinence and genital prolapse, but there were significant differences between the two study groups and the control group. Of the patients in the study group, 67 percent had urinary incontinence before or at the time of surgery. CONCLUSION: There is a higher prevalence and severity of urinary incontinence and pelvic genital prolapse in females operated on for either fecal incontinence or rectal prolapse than in a control group. Therefore, female patients with fecal incontinence or rectal prolapse should be evaluated and treated by a multidisciplinary group of pelvic floor clinicians, including a gynecologist or urologist with special training in female pelvic floor dysfunction and a colorectal surgeon. PMID- 11496070 TI - Hybrid laparoscopic flexure takedown and open procedure for rectal resection is associated with significantly shorter length of stay than equivalent open resection. AB - PURPOSE: Laparoscopic-assisted, sphincter-saving resection (largest incision < 7 cm) of the middle and distal rectum is technically very difficult and, with regard to cancers, has not been demonstrated to be oncologically safe. The hypothesis of this retrospective study is that a hybrid operation that combines laparoscopic and open methods would be associated with short-term outcome benefits compared with open surgery results for patients undergoing sphincter saving proctectomy. METHODS: A total of 31 hybrid and 25 fully open rectal resection patients were compared in this retrospective review. All patients had splenic flexure takedown and rectal anastomosis. The hybrid approach consisted of laparoscopic splenic flexure takedown (with or without partial rectal mobilization and devascularization) followed by completion of the procedure via infraumbilical midline laparotomy. The indication was neoplasm in 87 percent of hybrid patients and in 68 percent of open patients. The majority of tumors were located between 4 and 10 cm from the dentate line. RESULTS: Fifty-eight percent of hybrid and 68 percent of open patients had low anterior or coloanal resections, and 48 percent of hybrid and 64 percent of open patients underwent temporary diversion via ileostomy. The mean hybrid midline incision length was 11 cm compared with 24 cm for open patients (P < 0.0001). The neoplastic specimens were similar with regard to margins and lymph node harvest. Similar complication rates were noted in both groups. Nonsignificant benefits for hybrid patients (0.9 1.2 days) were seen with regard to length of time until toleration of liquid or solid diet and first flatus. Hybrid patients experienced their first bowel movements 4.1 days vs. 5.7 days for the open group (P = 0.03). Mean length of stay was significantly shorter for hybrid patients (6.1. days) than for open patients (11.1 days; P = 0.0006). CONCLUSION: This preliminary retrospective study suggests that a combined hybrid laparoscopic and open approach to sphincter saving proctectomy permits a similar resection as open methods and may be associated with a length-of-stay benefit and more rapid return of bowel function. Prospective studies will be needed before any firm conclusions can be drawn. PMID- 11496072 TI - Initial nonoperative management for periappendiceal abscess. AB - PURPOSE: Our goal was to compare initial operative and nonoperative management for periappendiceal abscess complicating appendicitis. METHODS: This study is a retrospective review of 155 consecutive patients with appendicitis complicated by periappendiceal abscess treated between 1992 and 1998. Eighty-eight patients were treated initially nonoperatively, and 67 patients were treated operatively. All patients had localized abdominal tenderness and either computed tomography or intraoperative documentation of an abscess. RESULTS: Our patient population consisted of 107 males and 48 females, with an average age of 33 (range, 16-75) years. Age, gender, comorbidity, white blood cell count, temperature, and heart rate did not differ significantly between groups. For the initial nonoperative management group, the failure rate was 5.8 percent and the appendicitis recurrence rate was 8 percent after a mean follow-up of 36 weeks. The response to treatment of the initial nonoperative group and the initial operative group was compared by length of stay (9 +/- 5 days vs. 9 +/- 3 days; P = not significant), days until white blood cell count normalized (3.8 +/- 4 days vs. 3.1 +/- 3 days; P = not significant), days until temperature normalized (3.2 +/- 3 days vs. 3.1 +/- 2 days; P = not significant), and days until a regular diet was tolerated (4.7 +/- 4 days vs. 4.6 +/- 3 days; P = not significant). Complication rate was significantly lower in the nonoperative group (17 vs. 36 percent; P = 0.008). CONCLUSIONS: Initial nonoperative management of appendicitis complicated by periappendiceal abscess is safe and effective. Patients undergoing initial nonoperative management have a lower rate of complications, but they are at risk for recurrent appendicitis. PMID- 11496073 TI - Readmissions after colorectal surgery cannot be predicted. AB - INTRODUCTION: Readmission after discharge from the hospital is an undesirable outcome. In an attempt to prevent unplanned readmissions after abdominal or perineal colon resection, we proposed to identify risk factors associated with return to the hospital. METHODS: Study participants consisted of 249 patients who were operated on from July 1, 1996, to March 30, 1998. All patients who were readmitted within 90 days of discharge from the hospital after surgery were evaluated for the study. A retrospective review of charts was performed to assess whether readmission within 90 days was a direct consequence of the recent operation (unplanned related readmission). These patients were compared with a control group consisting of patients who were never readmitted or who were readmitted with an unrelated problem. RESULTS: Of the 249 patients, 59 (24 percent) were readmitted within 90 days of discharge from the hospital. Twenty two (9 percent) were unplanned related readmissions. Ten patients were readmitted with unrelated emergencies, and 27 patients were readmitted electively. In the unplanned related group, there was no correlation between age, gender, admission diagnosis, activity status, or postoperative length of stay and the likelihood of readmission. Patients with multiple chronic medical problems or those who developed postoperative complications did not have a higher readmission rate. Patients with ulcerative colitis or those who underwent abdominoperineal resection or total/subtotal colectomy had a higher incidence of readmissions, although the difference was not significant. The mean interval between discharge from the hospital and readmission with a related complication was 19 days. Small bowel obstruction was the most common reason for readmission, and all cases resolved with conservative management. Mean length of stay during all readmissions was 8 days. CONCLUSION: The incidence of unplanned related readmissions 90 days after abdominal or perineal colon resection is 9 percent, and these readmissions could not be predicted from the postoperative course. Because 82 percent of unplanned readmissions occurred within 30 days, this time frame is suitable for computerized comparative analysis. PMID- 11496074 TI - Local recurrence after curative anterior resection with principally blunt dissection for carcinoma of the rectum and rectosigmoid. AB - PURPOSE: The aim of this study was to determine the incidence of local pelvic recurrence of carcinoma of the rectum and rectosigmoid (tumors where the lower edge is 18 cm or less from the anal verge) in a consecutive series of patients operated on by a single surgeon. All patients underwent curative anterior resection and a formal anatomic dissection of the rectum where mobilization was achieved through a principally careful blunt manual technique along fascial planes, preserving an oncologic package. METHOD: During the period April 1986 to December 1997, 157 consecutive anterior resections for carcinoma of the rectum and rectosigmoid were performed by one surgeon (ALP). One hundred thirty-eight (87.9 percent) were curative, and 19 (12.1 percent) were palliative. The mean follow-up period was 46 +/- 31.6 (range, 2-140) months. Data were retrospectively collated and computer coded by an independent contracted medical research team. Follow-up data were available on all patients. RESULTS: Four (3.1 percent) of the 131 patients undergoing curative anterior resection had local recurrence. Local recurrences occurred between 16 and 38 months from the time of resection, and the cumulative risk of developing local recurrence at five years was 5.2 percent. All tumors in which pelvic recurrence occurred were high grade, and the probability of developing local recurrence at five years for this group was 13.9 percent, which is significantly higher compared with patients who had average or low-grade tumors (P = 0.01). The probability of developing local recurrence at five years for Stage I tumors was 0, Stage II was 5.9 percent, and Stage III was 8.9 percent. In addition, there was a significantly higher incidence of local recurrence in the group of patients undergoing ultralow anterior resection (between 3 and 6 cm from the anal verge) as compared with patients undergoing low or high anterior resection (P = 0.03). Local recurrence developed in 3 of 28 (10.7 percent) patients having ultralow anterior resection, 1 of 57 (1.8 percent) patients having low anterior resection (between 6 and 10 cm from the anal verge), and no patients having high anterior resection (above 10 cm from the anal verge). The clinical anastomotic leak rate for curative anterior resection was 7 of 131 patients (5.3 percent). Thirty-seven of the 131 (28.2 percent) required a proximal defunctioning stoma; 35 (41.2 percent) of these were established for low or ultralow anterior resections and 2 for high anterior resection. The overall five-year cancer-specific survival rate of the entire group of 131 patients was 81.8 percent, and the overall probability of being disease free at five years including both local and distal recurrence was 72.9 percent. Three local recurrences occurred in the 101 patients (77 percent) who did not receive any form of adjuvant therapy. One local recurrence occurred in the 18 patients (13.7 percent) who had adjuvant chemoradiation. No recurrence occurred in the 12 patients (9.2 percent) who had adjuvant chemotherapy alone. CONCLUSION: Curative anterior resection for carcinoma of the rectum and rectosigmoid with principally blunt dissection of the rectum in this study is associated with a 3.1 percent incidence and a 5.2 percent probability at five years of developing local recurrence. Evidence from this study indicates that, as with sharp pelvic dissection, a low incidence and probability of local recurrence can be achieved by a principally blunt mobilization technique through careful attention to preservation of fascial planes in the pelvis and removal of an oncologic package with selective rather than routine adjuvant or neoadjuvant chemoradiation. PMID- 11496075 TI - Storage time of transfused blood and disease recurrence after colorectal cancer surgery. AB - BACKGROUND: Perioperative blood transfusion and subsequent development of postoperative infectious complications may lead to poor prognosis of patients with colorectal cancer. It has been suggested that the development of postoperative infectious complications may be related to the storage time of the transfused blood. Therefore, we studied the relationship between blood storage time and the development of disease recurrence and long-term survival after colorectal cancer surgery. METHODS: Preoperative and postoperative data were prospectively recorded in 740 patients undergoing elective resection for primary colorectal cancer. None of the patients received preoperative or postoperative chemotherapy or radiation therapy. Endpoints were overall survival and disease recurrence in the subgroup of patients operated on with curative intention who also survived the first 30 days after operation. Storage of buffy-coat-depleted red cells suspended in saline, adenine, glucose, and mannitol blood for 21 days was used as cut-off point. RESULTS: Median follow-up was 6.8 years (range, 5.4 years to 7.9 years), and median overall survival was 4.6 years for 288 nontransfused patients and 3.0 years for 452 transfused patients (P = 0.004). The survival of patients receiving blood exclusively stored < 21 days was 2.5 years. For patients receiving any blood stored > or = 21 days, survival was 3.7 years (P = 0.12). Among patients with curative resection (n = 532), the hazard ratio of disease recurrence was 1.5 (95 percent CI; 1.1 to 2.2) and 1.0 (95 percent CI; 0.7 to 1.4) in the two transfused groups, respectively, compared with the nontransfused group after multivariable correction for patient age, gender, colonic/rectal tumor localization, Dukes classification, blood loss, and postoperative infectious complications. CONCLUSION: Transfusion of buffy-coat depleted red cells suspended in saline, adenine, glucose, and mannitol blood stored for < 21 days may be an independent risk factor for development of recurrence after elective colorectal cancer surgery. PMID- 11496076 TI - Neuromodulation for fecal incontinence: outcome in 16 patients with definitive implant. The initial Italian Sacral Neurostimulation Group (GINS) experience. AB - PURPOSE: Sacral nerve modulation appears to offer a valid treatment option for some patients with fecal incontinence and functional defects of the internal anal sphincter or of the striated muscle. METHODS: Sixteen patients with fecal incontinence (4 males; mean age, 51.4 (range, 27-79) years) with intact or surgically repaired (n = 1) anal sphincter underwent permanent sacral nerve stimulation implant. Cause was traumatic in two patients, and associated disorders included scleroderma (2 patients) and spastic paraparesis (1 patient); eight (50 percent) of the patients also had urinary incontinence, and two (12.5 percent) had nonobstructive urinary retention. All patients were selected on the basis of positive findings from at least one peripheral nerve evaluation. The stimulating electrode was positioned in the S2 (1 patient), S3 (14 patients), or S4 (1 patient) sacral foramen. RESULTS: Mean follow-up was 15.5 (range, 3-45) months. Mean preimplant Williams score decreased from 4.1 +/- 0.9 (range, 2-5) to 1.25 +/- 0.5 (range, 1-2) (P = 0.01, Wilcoxon test), and the number of incontinence accidents for liquid or solid stool in 14 days decreased from 11.5 +/- 4.8 (range, 2-20) before implant to 0.6 +/- 0.9 (range, 0-2) at the last follow-up. Important manometric data were an increase in mean maximal pressure at rest of 37.7 +/- 14.9 mmHg (implantable pulse generator 49.1 +/- 18.7, P = 0.04) and in mean maximal pressure during squeeze (prestimulation 67.3 +/- 21.1 mmHg, implantable pulse generator 82.6 +/- 21.0, P = 0.09). CONCLUSIONS: Neuromodulation can be considered an option for fecal incontinence. However, an accurate clinical and instrumental evaluation and careful patient selection are required to optimize outcome. PMID- 11496077 TI - Rectal compliance in females with obstructed defecation. AB - PURPOSE: This study was designed to investigate whether rectal compliance is altered in females with obstructed defecation. METHODS: Eighty female patients with obstructed defecation and 60 control subjects were studied. Rectal compliance was measured with an "infinitely compliant" polyethylene bag. This bag was inserted in the rectum and inflated with air to selected pressure plateaus (range, 0-60 mmHg; cumulative steps of 2 mmHg with a duration of ten seconds) using a computer-controlled electromechanical barostat system. Volume changes at the levels of distending pressures were recorded. The distending pressures, needed to evoke first sensation of content in the rectum, earliest urge to defecate, and the maximum tolerable volume were noted. RESULTS: In all cases, the compliance curve had a characteristic triphasic (S-shaped) form. The mean compliance curve obtained from the patients was identical to that of the controls. However, the course of the compliance curve fell above the normal range (mean + 2 SD) in 14 patients. In ten (71 percent) of these patients, a large rectocele was seen at evacuation proctography. Such a rectocele was observed in only five patients (7.6 percent) with a normal compliance curve (P < 0.001). Eighty percent of the controls experienced earliest urge to defecate during the second phase of the curve. In 75 percent of the patients, this occurred in the third phase. The mean pressure threshold for first sensation, earliest urge to defecate, and maximum tolerable volume were significantly higher in patients compared with control subjects. Ten of the patients experienced no sensation at all in the pressure range between 0 and 60 mmHg. CONCLUSION: In females with obstructed defecation, the compliance of the rectal wall is normal. PMID- 11496078 TI - Inadequacy of colonoscopy revealed by three-dimensional electromagnetic imaging. AB - INTRODUCTION: It is generally accepted that clinicians performing colonoscopy should reach the cecum in at least 90 percent of examinations. However, little attention has been paid to whether the endoscopist correctly estimates the amount of colon actually seen. METHODS: During colonoscopy, endoscopists were asked to state how far they had reached. This was compared with the amount of colon actually seen, as assessed by a novel electromagnetic imaging device that recorded a three-dimensional position of the scope within a magnetic field pervading the patient's abdomen. If electromagnetic imaging showed that the cecum had not been reached, the endoscopist was asked to use the electromagnetic imaging system to determine whether it helped advance the colonoscope further. RESULTS: In 119 patients undergoing colonoscopy, clinical assessment of position reached was correct in only 92 (77.3 percent). When the endoscopists stated that cecal landmarks had been seen (n = 85), the scope was distal to the cecum in seven cases (8.2 percent). When cecal landmarks had not been seen (n = 34), the endoscopist's assessment of the position of the scope was accurate in only 14 (41.2 percent). The use of electromagnetic imaging in this latter group assisted passage to the cecum in 26 cases (76.5 percent). CONCLUSION: Despite assumed visualization of the cecum, inadequate colonoscopy highlights the potential for missing significant pathology in the right colon. PMID- 11496079 TI - Outcome of primary and secondary ileal pouch-anal anastomosis and ileorectal anastomosis in patients with familial adenomatous polyposis. AB - PURPOSE: The aim of this study was to present Swedish experiences of the ileal pouch-anal anastomosis in patients with familial adenomatous polyposis from the introduction in 1984. The study also compared the surgical and functional outcome of different anal continence preserving procedures: ileal pouch-anal anastomosis as primary surgery, ileal pouch-anal anastomosis as secondary surgery after colectomy and ileorectal anastomosis, and ileorectal anastomosis alone. METHODS: The material comprises all 120 patients with familial adenomatous polyposis reported to the Swedish Polyposis Registry who had undergone prophylactic colorectal surgery, including those operated on because of colorectal cancer from 1984 until the end of 1996. Anal continence preserving surgery was performed on 102 patients: 20 had ileal pouch-anal anastomosis as primary surgery at a median age of 24.5 years, 39 had ileal pouch-anal anastomosis as secondary surgery at a median age of 34 years, and 43 had ileorectal anastomosis alone, at a median age of 26 years, because 6 of the initially ileorectal anastomosis-operated patients were converted to ileal pouch-anal anastomosis as secondary surgery. Surgical outcome was assessed on the basis of hospital records. A questionnaire was used to evaluate the functional outcome. Fisher's exact probability test was used for statistical analysis. RESULTS: Complications occurred in 51 percent of the patients after ileal pouch-anal anastomosis: 40 percent after ileal pouch-anal anastomosis as primary surgery and 56 percent after ileal pouch-anal anastomosis as secondary surgery. When the previous ileorectal anastomosis was taken into account 67 percent of the patients suffered complications which was significantly more compared with ileal pouch-anal anastomosis as primary surgery. After ileorectal anastomosis, 26 percent had complications which was significantly less compared with all other procedures but ileal pouch-anal anastomosis as primary surgery. No cancer occurred after ileal pouch-anal anastomosis, either in the ileal pouch or in retained rectal mucosa, but two of the patients who had an ileorectal anastomosis developed rectal cancer. One pouch excision was performed compared with ten rectal excisions. Functional outcome did not differ between ileal pouch-anal anastomosis as primary surgery and ileal pouch-anal anastomosis as secondary surgery. However, ileorectal anastomosis-operated patients had significantly better bowel function with regard to nighttime stool frequency, continence and perianal soreness. CONCLUSION: These findings indicate that major advantages of ileal pouch-anal anastomosis are the low excision rate and, so far, no cancer in the ileal pouch. Moreover, the surgical outcome of ileal pouch-anal anastomosis as primary surgery is not significantly different from that of ileorectal anastomosis. However, the good surgical and functional outcome of ileorectal anastomosis, despite the long-range prognosis including rectal cancer and excision risks, has to be taken into consideration when selecting patients with familial adenomatous polyposis for primary surgery. PMID- 11496080 TI - Cell-mediated immune response in megacolon from patients with chronic Chagas' disease. AB - PURPOSE: The mechanisms that control chronic infection in vivo and the immunologic mechanisms involved in the pathogenesis of chagasic megacolon are not completely characterized. Although autoimmunity may play a role in the pathogenesis of Chagas' disease, recent studies, both in mice and in humans, suggest a positive association of tissue parasitism, inflammation, and severity of lesions. The aim of this study was to evaluate the role of inflammatory cells and the subclasses of lymphocytes involved in neuropathic lesions in the colon of patients who underwent resection for advanced megacolon. METHODS: Specimens from 23 patients were selected based on histopathologic analysis. Paraffin-embedded tissue blocks were sectioned and evaluated by immunohistochemistry for cluster of differentiation 3, cluster of differentiation 8, cluster of differentiation 20, and natural killer cell antibodies by an avidin-biotin peroxidase method. RESULTS: Almost all myenteric plexuses were damaged, characterized by degenerative changes, necrosis of ganglion cells, and inflammatory response. Mild lymphocytic infiltration around degenerated and normal ganglion cells was observed in all cases. Collagen fibers and mononuclear cells surrounded some ganglion cells. Most of the inflammatory cells were lymphocytes, identified as cluster of differentiation 3-positive cells. Cluster of differentiation 8 positive lymphocytes were associated with degenerated ganglion cells. Natural killer cell antibodies were detected in a lower proportion of cells and were distributed between muscle layers or in proximity to the myenteric plexus. All these findings were also observed in the submucosal plexus. Cluster of differentiation 20-positive lymphocytes were not present in muscle layers or in the vicinity of either plexus. CONCLUSION: Pathogenesis of the megacolon is based on a continuous process of ganglion cell damage with participation of T lymphocytes expressing cluster of differentiation 8 and natural killer cell membrane antigens. B lymphocytes do not take part in the chronic inflammatory reaction. PMID- 11496081 TI - Dynamic magnetic resonance imaging defecography: a diagnostic alternative in the assessment of pelvic floor disorders in proctology. AB - PURPOSE: Standard diagnostic proctologic procedures in the assessment of pelvic floor disorders include clinical evaluation and endoscopy. Particular aspects of combined pelvic floor disorders, especially those involving more than one pelvic compartment, may remain undetected without additional technical diagnostic procedures such as videoproctoscopy, cinedefecography, or colpocystodefecography. The aim of the study was to review the potentials of dynamic magnetic resonance imaging defecography to elucidate the underlying anatomic and pathophysiologic background of pelvic floor disorders in proctologic patients. PATIENTS AND METHODS: Dynamic magnetic resonance imaging defecography was performed in 20 Patients (13 females) with main diagnoses such as rectal prolapse or intussusception, rectocele, descending perineum, fecal incontinence, outlet obstruction, and dyskinetic puborectalis muscle after clinical evaluation. The investigation was performed on a 1.5 T-magnetic resonance imaging machine in supine position. The rectum was filled with Gd-DTPA enriched ultrasound gel. First a T1/T2 weighted investigation of the pelvis was performed, followed by defecography with evacuation of the rectum. Images were obtained in a sagittal plane in a frequency of 1 image/second (true FISP) at rest and during straining. The obtained magnetic resonance imaging video tapes were analyzed off-line with cinematographic evaluation of bladder base, uterus, and anal canal position in relation to the pubococcygeal line by a blinded radiologist. Investigation time was 20 minutes. RESULTS: In dynamic magnetic resonance imaging defecography of the pelvic floor, 12 patients with descending perineum, 10 rectoceles (10 females), 6 cystoceles (6 females), 4 enteroceles (4 females), 8 intussusceptions (5 females), and a dyskinetic puborectalis muscle in 3 males were detected. In 11 females and 3 males multifocal disorders were found, involving more than one compartment in females, whereas in males complex defects were restricted to the posterior compartment. Magnetic resonance imaging defecography revealed diagnoses consistent with clinical results in 77.3 percent and defects in addition to clinical diagnoses in combined pelvic floor disorders in 34 percent. CONCLUSIONS: In complex pelvic floor disorders, involving more than a single defect, dynamic magnetic resonance imaging represents a convenient diagnostic procedure in females and to a lesser extent in males, in particular in terms of dynamic imaging of pelvic floor organs during defecation. In addition to the clinical assessment, dynamic magnetic resonance imaging had clinical impact in proctologic and interdisciplinary treatment. PMID- 11496082 TI - Enhanced apoptosis and transforming growth factor-beta1 expression in colorectal adenomas and carcinomas after Sulindac therapy. AB - PURPOSE: We tried to elucidate the effects of sulindac on human colorectal carcinoma. METHODS: Sulindac (300 mg/day) was administered for two weeks before operation to 33 patients with sporadic colorectal carcinoma (Sulindac Group). Resected specimens were used to detect apoptosis by terminal dUTP nick end labeling and transforming growth factor (TGF)-beta1 expression by immunohistochemistry. The results were compared with those from the historical Control Group. Twenty-nine available preoperative biopsies taken from carcinomas before sulindac prescription and 22 concurrent colorectal adenomas (9 and 13 in Sulindac and Control Groups, respectively) in the resected specimen were also examined regarding TGF-beta1 expression. RESULTS: In the resected carcinomas and adenomas, more frequent apoptosis and higher TGF-beta1 scores were observed in the Sulindac Group than in the Control Group. Overexpression of TGF-beta1 and apoptosis occurred in the same region in adenomas but not in carcinomas. A positive correlation between TGF-beta1 scores and apoptotic frequency was found in adenomas (P = 0.01, rho = 0.91) but not in carcinomas (P = 0.89, rho = 0.03). CONCLUSION: We conclude that sulindac induces apoptosis in human colorectal carcinomas as well as in adenomas. Also, one of the antineoplastic effects of sulindac might be mediated by upregulating TGF-beta1 expression, particularly in colorectal adenomas. PMID- 11496083 TI - Dietary fiber and colorectal neoplasia. AB - PURPOSE: Dietary fiber has been implicated in colorectal neoplasia, despite conflicting evidence. This is a review of the currently available data on the role of dietary fiber in colorectal carcinogenesis. METHODS: A literature search was conducted using the MEDLINE database. All case-control, longitudinal, and randomized, controlled studies published in English between 1988 and 2000 were identified, as were animal model studies in the period 1986 to 2000. Data from the various studies were tabulated and systematically analyzed, with particular emphasis on the effect of dietary fiber on tumor incidence and luminal parameters such as short chain fatty acids. RESULTS: Epidemiologic correlation studies show a high intake of dietary fiber to be associated with a lower risk of colorectal neoplasia. Thirteen of the 24 case-control studies reviewed demonstrated a protective effect of dietary fiber against colorectal neoplasia, and 16 showed a protective effect of vegetables or vegetable fiber. On the other hand, of 13 longitudinal studies in various cohorts, only 3 demonstrated a protective effect of fiber and 4 a protective effect of vegetables or vegetable fiber. The five published randomized, controlled trials all investigated the effect of increased fiber intake on short-term adenoma recurrence; however, none showed any significant protective effect. Among 19 experimental studies in animal models, 15 showed a protective effect of fiber against tumor induction compared with controls. Animal studies also showed that poorly fermentable fibers (e.g., wheat bran and cellulose) were more protective than soluble fibers (e.g., guar gum and oat bran), which sometimes enhanced carcinogenesis. No clear correlation was found between luminal pH or short chain fatty acids and tumor induction. CONCLUSIONS: On the basis of current data, there is little evidence to support the use of dietary fiber supplements to reduce the risk of colorectal neoplasia. Lifelong and early exposure may be important but are difficult to study. Other risk factors interact with the effects of dietary fiber. PMID- 11496084 TI - Giant mucocele of the appendix: report of a case. PMID- 11496085 TI - Penetrating colon ulcer of polyarteritis nodosa: report of a case. AB - A 54-year-old Japanese female with polyarteritis nodosa was admitted to the hospital. She developed lower abdominal pain accompanied by melena. A penetrating ulcer and extensive hemorrhaging were endoscopically observed in the sigmoid colon, and a sigmoidectomy was performed. The pathologic findings were a granuloma formation with lymphocytic infiltration and luminal occlusion of branches of the mesenteric arteries. Although the gastrointestinal tract is frequently involved in polyarteritis nodosa, the colon is rarely affected. To our knowledge, this is the first report of polyarteritis nodosa causing a penetrating ulcer of the colon. PMID- 11496086 TI - Cystic lymphangioma of the colon: endosonographic diagnosis with through-the scope catheter miniprobe and determination of further management. Report of a case. AB - Lymphangioma of the colon is rare. There are several reports that endoscopic ultrasound is useful for diagnosis of colonic lymphangioma. We report a case of lymphangioma of colon diagnosed by catheter endosonography and review the literature on endoscopic ultrasound in cystic lymphangioma of the gastrointestinal tract. A 70-year-old female was found to have two submucosal lesions in the colon by colonoscopy. Endoscopic ultrasound revealed that these lesions were anechoic, multicystic, and confined to the submucosa, and the underlying muscularis propria was intact. These findings were consistent with cystic lymphangioma. If typical endosonographic images of an anechoic, septated lesion within colonic submucosa are obtained, further workup or treatment may not be necessary if the patient is asymptomatic. PMID- 11496087 TI - Rectal cancer in anorectal malformation with rectovestibular fistula: sphincter restoring surgery by anterior resection and dynamic vaginoanograciloplasty. AB - PURPOSE: The aim of this study was to describe and discuss the anosphincter restoring procedure adopted in an adult patient with rectal cancer, affected by anorectal atresia and rectovestibular fistula not previously treated. METHODS: After anterior resection and transverse dissection of the rectovaginal septum, a straight double-staple low colorectal anastomosis was performed. A subcutaneous tunnel was extended from the inter-rectovaginal space, surrounding the pseudoanal orifice. The tunnel housed a left gracilis muscle arranged counter-clockwise and connected to an implanted stimulator. The vaginal and anorectal flaps were sutured longitudinally, thereby spacing out the two orifices and creating an anorectal angle. A temporary loop right colostomy was performed. RESULTS: The postoperative course was uneventful. After continuous electrostimulation training and colostomy takedown, the patient had improved continence (stimulator on) and normal defecation (stimulator off). CONCLUSIONS: The solution described seemed a suitable alternative to a pull-through procedure, which would have involved a colocutaneous anastomosis and the loss of the anorectal sensory function. Any previous anal transposition or pull-through procedures would have prevented the saving of the pre-existing "anus," because of the interruption of the residual mesenteric vascular supply to the anorectal remnant secondary to mesorectal excision. PMID- 11496088 TI - Colonoscopy without sedation. PMID- 11496089 TI - Fecoflowmetry with artificial stools vs. dynamic videoproctography. PMID- 11496090 TI - Glucose alters the susceptibility of mesangial cells to contrast media. AB - RATIONALE AND OBJECTIVES: Diabetic patients frequently suffer contrast media induced nephropathy. Hyperglycemia in diabetes mellitus causes gradual deterioration of glomerular mesangial cells (MCs) in the kidney. In this study, the authors investigated the response of rat MCs cultured in high-glucose medium to diatrizoate and iohexol, high- and low-osmolar contrast media, respectively. METHODS: Isolated rat MCs were precultured under basal-glucose (5.5 mmol/L) and high-glucose (30 and 55 mmol/L) conditions for 24 hours to mimic hyperglycemia in diabetes mellitus and then were exposed to diatrizoate (40 and 80 mg I/mL) and iohexol (80, 120, 160 mg I/mL) for 2 hours. The cytotoxic effects of diatrizoate and iohexol were monitored by neutral red uptake in MCs. The protective effects of an antioxidant, d-alpha-tocopherol (Toc), on cytotoxicity of the contrast media were determined when MCs were precultured with Toc under high-glucose conditions or were exposed to the contrast media together with Toc. Peroxide levels in the cells exposed to the contrast media were analyzed by flow cytometry using dichlorofluorescin diacetate. RESULTS: Exposure to both contrast media (diatrizoate and iohexol) induced a concentration-dependent decrease in viability of the cells precultured under basal-glucose conditions (5.5 mmol/L). Preculture under high-glucose conditions (30 and 55 mmol/L) augmented the cytotoxic effects of both contrast media. An increase in the intracellular peroxide level was detected after exposure to both contrast media. Preculture with Toc prevented augmentation of the cytotoxic effects of diatrizoate by the higher glucose concentration (55 mmol/L). The exposure to diatrizoate together with Toc also attenuated its cytotoxic effects. Toc showed no such protective effects against iohexol exposure. CONCLUSIONS: These findings suggest that high-glucose conditions enhance the susceptibility of MCs to the cytotoxic effects of both contrast media; the enhanced susceptibility was in part attributable to oxidative stress caused by high-glucose conditions; diatrizoate exerted the cytotoxic effects by means of oxidative stress; and iohexol appeared to exert its cytotoxicity in a manner different from diatrizoate. PMID- 11496091 TI - Improved reproducibility of coronary artery calcium scoring by electron beam tomography with a new electrocardiographic trigger method. AB - RATIONALE AND OBJECTIVES: To improve the interscan reproducibility with electron beam tomography (EBT) by choosing an optimal electrocardiographic (ECG) trigger time. METHODS: Two hundred fourteen asymptomatic subjects found to have coronary artery calcium (CAC) on EBT were rescanned immediately to measure the interscan variability. Subjects were randomized to one of two different ECG trigger interval groups: the new trigger method (group 1) and the 80% R-R interval trigger method (group 2). The new trigger method was derived from a previous study of motion in the coronary arteries. In group 1 (new trigger method), the ECG trigger was programmed for a certain time (in ms) after the R wave, based on the resting heart rate. The triggers for group 1 were 360 (heart rate <50 beats per minute [bpm]), 340 (51--60 bpm), 314 (61--70 bpm), 300 (71--80 bpm), 290 (81- 90 bpm), 280 (91--100 bpm), and 270 ms (>100 bpm). The interscan variation (CAC area and Agatston score) was compared between the two groups. RESULTS: The interscan variability was significantly reduced using the new trigger method for both CAC area and score compared with the 80% trigger method. The individual lesion variation was also significantly reduced by the new trigger method compared with the 80% trigger method. Area measure had a significantly lower variability compared with the Agatston score. CONCLUSIONS: These results strongly support the use of this new ECG trigger that relies on a rate-adjusted millisecond delay after the R wave instead of the more commonly used 80% R-R interval in EBT calcium studies. PMID- 11496092 TI - Cerebrospinal fluid dynamics and relation with blood flow: a magnetic resonance study with semiautomated cerebrospinal fluid segmentation. AB - RATIONALE AND OBJECTIVES: To investigate and measure temporal and amplitude aspects of blood and cerebrospinal fluid (CSF) flow waveform relations. METHODS: A cine phase-contrast magnetic resonance imaging pulse sequence was used to measure blood and CSF flow in 16 healthy subjects aged 27 +/- 4 years. A semiautomated segmentation algorithm was developed to study CSF flow. RESULTS: Standard deviations of the aqueductal and cervical flow measurements carried out by five observers were 1% and 4%, respectively. The peak systolic arterial flow was 1087 +/- 169 mL/min, and the peak cervical CSF flush (173 +/- 59 mL/min) occurred at 5% +/- 3% of the cardiac cycle after the internal carotid systolic peak flow. Peak aqueductal flush flow (13 +/- 5 mL/min) occurred at 21% +/- 7% of the cardiac cycle after the internal carotid systolic peak flow. CONCLUSIONS: The CSF segmentation algorithm is reproducible. Brain expansion was quickly regulated by a major extracerebral CSF flush flow, whereas ventricular CSF made only a very small contribution. PMID- 11496093 TI - Perfusion MRI of infarcted and noninfarcted brain tissue in stroke: a comparison of conventional hemodynamic imaging and factor analysis of dynamic studies. AB - RATIONALE AND OBJECTIVES: To investigate the hemodynamics of infarcted and noninfarcted regions of the brain in patients with stroke secondary to a complete middle cerebral artery occlusion. Also, to compare factor analysis, a novel method of analyzing perfusion-weighted images, with more conventional techniques. METHODS: Twenty-two patients with complete unilateral occlusion of the middle cerebral artery were examined by T1-weighted, contrast-enhanced, perfusion weighted imaging, diffusion-weighted imaging, and magnetic resonance angiography. Quantitative cerebral blood volume (CBV), cerebral blood flow (CBF), and time-to peak-intensity (TTP) images were generated. Factor analysis of dynamic studies (FADS) was used to generate "early" and "late" images. The hemodynamic parameters for the infarcted and noninfarcted regions of the occluded territory were compared with those for the brain territory on the nonoccluded side. RESULTS: Three regions were shown: (1) Normal tissue on the unaffected side; (2) an infarcted region, which was characterized by reduced CBV, CBF, and early FADS values with increased TTP values; and (3) a noninfarcted region with reduced early FADS and increased late FADS and TTP values compared with the normal region. Cerebral blood volume and CBF values were not reduced significantly in the noninfarcted region. CONCLUSIONS: The differences in parameters such as TTP, CBV, and CBF are significant, and it is necessary to use more than one parameter when interpreting magnetic resonance imaging perfusion data. Factor analysis of dynamic studies provides additional information to conventional methods of analyzing perfusion data. PMID- 11496094 TI - Perfusion characteristics of oleic acid--injured canine lung on Gd-DTPA--enhanced dynamic magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: We conducted an animal study to describe and interpret the perfusion characteristics of oleic acid (OA)-injured lungs on gadopentetate dimeglumine (Gd-DTPA)-enhanced dynamic perfusion magnetic resonance (MR) imaging. METHODS: Fourteen dogs received an intravenous OA infusion in the supine (n = 4), prone (n = 4), and right lateral decubitus (n = 6) positions, and 10 minutes later these animals in the same postures underwent the dynamic MR study. Regional Gd-DTPA kinetics was analyzed by the time-signal intensity (SI) curves and by qualitative functional map images of the mean transit time that was representative of the mean circulation time in the vascular bed and the average cumulative sum of the relative increases in SI representative of Gd-DTPA distribution volume during Gd-DTPA first pass. The results were compared with those in six control animals and in another six animals that underwent the MR study 3 minutes (n = 3) and 60 minutes (n = 3) after OA infusion. The MR findings were correlated with the distribution of lung damage and the infused OA particles as assessed by histology. RESULTS: The dynamic MR study showed postural shifts on the gravity-dependent perfusion map of normal lungs. Contrast enhancement during Gd-DTPA first pass in the lung was lower and more heterogeneous in the OA-injured lung models than in controls but was followed by conversely greater and persistent enhancement during the Gd-DTPA redistribution phase. Regardless of the postures for OA infusion, these abnormalities were predominant in the dependent lungs and became more pronounced with time after OA infusion, where more prominent capillary obstruction with OA droplets and alveolar/interstitial edema were histologically observed. On the functional map images, greater mean transit time and the average cumulative sum of the relative increases in SI values were also predominantly distributed in the dependent lungs. CONCLUSIONS: Low and heterogeneous enhancement was observed during Gd-DTPA first pass but was followed by persistent enhancement during the Gd-DTPA redistribution phase, and predominant abnormalities in the dependent lungs may be characteristic features of the perfusion of OA-injured lungs. The histological correlations indicate that these abnormalities may reflect OA-induced pathophysiologies associated with capillary OA obstruction, increased vascular resistance, and capillary permeability/extravascular spaces and that lung damage may be gravity dependent. PMID- 11496095 TI - Toxicological assessment of gadoversetamide injection (OptiMARK), a new contrast enhancement agent for use in magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: A series of preclinical tests were undertaken during the developmental process to determine the safety profile of gadoversetamide injection (OptiMARK). METHODS: Acute intravenous, acute intracisternal, and repeated-dose toxicities; cardiovascular effects; and genetic and reproductive toxicology characteristics were assessed in several animal species. RESULTS: Gadoversetamide injection demonstrated an acute intravenous median lethal dose of 25 to 28 mmol/kg and a maximum nonlethal dose of 14 mmol/kg in mice. In the dog, acute administration of gadoversetamide injection showed a no observable effect level at 3 mmol/kg. Dosed daily for 4 weeks, gadoversetamide injection (0.1 mmol x kg(-1) x d(-1)) caused no serious irreversible changes in any organs in rats and dogs. At a dose of 0.1 mmol/kg, gadoversetamide injection caused no significant (P < 0.05) changes in cardiovascular function in anesthetized dogs. Gadoversetamide injection showed no mutagenic activity. Fertility, reproductive performance, and postnatal fetal development were not affected at doses up to 0.5 mmol x kg(-1) x d(-1) in the rat. No teratogenicity was observed at doses up to 4.2 mmol x kg(-1) x d(-1) in the rat and up to 1.6 mmol x kg(-1) x d(-1) in the rabbit. CONCLUSIONS: Data from our toxicological assessment demonstrate the safety of gadoversetamide injection in a number of animal species at doses exceeding the intended human clinical dose. PMID- 11496096 TI - Histopathological correlation to MRI findings during and after laser-induced thermotherapy in a pig pancreas model. AB - RATIONALE AND OBJECTIVES: To investigate whether percutaneous laser-induced thermotherapy (LITT) with continuous magnetic resonance (MR) monitoring of thermal effects within the pancreas is feasible in a porcine model. METHODS: Laser applicators were placed in the pancreas of 15 female pigs. A temperature sensitive (thermo--fast low-angle shot) sequence was used for continuous monitoring of thermal effects during LITT at 1.5 T. Follow-up MR images were acquired, the pigs were observed for 7 days, and then a pathological examination was performed after sacrifice. RESULTS: Continuous MR monitoring visualized thermal effects in pancreatic tissue and thermal damage of the spleen (n = 1), the left kidney (n = 1), and peripancreatic fat (n = 4) but missed the thermal damage of the duodenum (n = 2). Thermal-induced lesions (10--32-mm diameter) were clearly visualized on contrast-enhanced T1-weighted images. CONCLUSIONS: Laser induced thermotherapy of pancreatic tissue was feasible in this porcine model, and online monitoring was practicable. Further studies are necessary to increase the accuracy of online MR imaging of thermal effects. PMID- 11496097 TI - Cross-sectional pattern of collateral vessels in patients with superficial femoral artery occlusion. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to identify the cross sectional location of collateral vessels in patients with peripheral vascular disease on three-dimensional magnetic resonance angiograms (3D MRAs) to suggest sites for intravascular or transcutaneous angiogenesis gene delivery in the lower extremity. METHODS: The axial locations were measured and categorized by tissue compartments, as well as by radial coordinates with respect to the femur. RESULTS: Collateral vessels in the thigh were identified in 24 of 93 consecutive patients who underwent peripheral 3D MRA. Ninety-one percent (99/109) of the observed collaterals were located near the adductor canal level of the thigh, with 78% (31/46) of these collaterals located in the fat in or surrounding the posterior muscle. CONCLUSIONS: The majority of collateral vessels in the thigh are located in the fat or muscle within the posterior compartment near the femur at the level of the adductor canal. PMID- 11496099 TI - Conditioned blocking is re-established by neurotransplantation in mature rats. AB - Aging has been associated with a decay of hippocampal function that may begin well before senescence. Conditioned blocking is a complex learning phenomenon that requires an intact hippocampus in young-adult rats and is absent in middle aged rats. The aim of the present study was to test the possibility of re establishing conditioned blocking in 17-month-old Wistar rats by neurotransplantation. Solid embryonic hippocampal or nigral tissue was bilaterally transplanted in the proximity of the dorsal hippocampus (lateral ventricle and alveus). Conditioned blocking of an aversion to a cider vinegar (3%) solution presented in compound with a previously conditioned saccharin solution (0.1%) appeared 14 days after transplantation and persisted 3 months later only in the hippocampal grafted group, showing the possibility of restoring age-related cognitive deficits. PMID- 11496100 TI - Effects of ensheathing cells transplanted into photochemically damaged spinal cord. AB - Transplantation of olfactory ensheathing cells (OECs) into photochemically damaged rat spinal cord diminished astrocyte reactivity and parenchyma cavitation. The photochemical lesion performed at T12--L1 resulted in severe damage to the spinal cord, so that during the first 15 days postoperation all rats dragged their hindlimbs and did not respond to pinprick. The maximal area and volume of the cystic cavities were lower in transplanted than in non transplanted rats, not significantly at the T12--L1 lesion site, but significantly at T9--T10 and L4--L6 cord levels. The density of astrocytes in the grey matter was similar at T12--L1 and L4--L6 in non-transplanted and trans- planted rats, but lower in the latter at T9--T10 level. However, in non transplanted rats all astrocytes showed a hypertrophied appearance, with long and robust processes heavily GFAP-positive, and overexpression of proteoglycan inhibitor of neuritogenesis, whereas in transplanted rats only a few astrocytes showed hypertrophy and the majority had short, thin processes. These results indicate that OECs transplanted into damaged adult rat spinal cord exert a neuroprotective role by reducing astrocytic gliosis and cystic cavitation. PMID- 11496101 TI - Basal expression of bradykinin B(1) receptor in the spinal cord in humans and rats. AB - The bradykinin B(1) receptor has been considered as a receptor induced by tissue injury and inflammation mainly in the peripheral tissues. In the present study, we have investigated whether there is a basal expression in the spinal cord by both reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical staining methods. Southern blotting of the DNA reverse transcribed from human and rat spinal cord mRNA and amplified by polymerase chain reaction (PCR) showed a substantial basal B(1) receptor expression in both human and rat spinal cord. Immunohistochemical staining demonstrated B(1)-positive neurons in the spinal cord dorsal horn, suggesting that the B(1) receptor is constitutively expressed by spinal neurons. PMID- 11496102 TI - (1)H-MR spectroscopy differentiates mild cognitive impairment from normal brain aging. AB - This study aimed to characterize the white matter biochemical profile of healthy elderly subjects, mild cognitive impairment (MCI) subjects, and early Alzheimer's disease (AD) patients. We used proton magnetic resonance spectroscopy ((1)H-MRS) to measure myo-inositol, creatine, N-acetylaspartate (NAA) and choline levels from a volume of interest located in the paratrigonal white matter bilaterally. A significantly higher myo-inositol/creatine ratio was found in MCI subjects and AD patients than in controls. The NAA/creatine ratio was reduced in AD patients in the left hemisphere compared to control subjects. The choline/creatine ratio was not significantly different among the three groups. These data suggest that MCI is different from normal brain aging, having a white matter biochemical pattern similar to AD. PMID- 11496103 TI - Mutation screening of manganese superoxide dismutase in amyotrophic lateral sclerosis. AB - Seventy-seven cases of ALS were screened for mutations in the manganese superoxide dismutase gene (SOD2). DNA was extracted from CNS tissue and screened using single stranded conformation polymorphism and heteroduplex analysis. No mutations were identified in the entire coding region of the SOD2 gene. The known polymorphism in the mitochondrial targeting sequence was identified. No association was found between this polymorphism and ALS. A further polymorphism was detected in the intronic sequence upstream of exon 4, though no association with ALS was demonstrated. We therefore conclude that mutations in SOD2 do not appear to cause ALS. PMID- 11496104 TI - Tyrosine hydroxylase, but not dopamine beta-hydroxylase, is increased in rat frontal cortex after traumatic brain injury. AB - Chronic frontal lobe functional deficits after traumatic brain injury (TBI) may be associated with altered catecholamine systems in the frontal cortex. To test this, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) levels were examined by immunohistochemistry and Western blot at 1, 7, 14, and 28 days after TBI or sham surgery. No alterations in DBH levels were observed by Western blot at any time point examined, but there was a significant increase in TH expression 28 days after TBI (optical density 334 +/- 68% or 3.3-fold, ipsilateral and 218 +/- 39% or 2.2-fold, contralateral) relative to the sham controls. The increase in TH may reflect a compensatory response of dopaminergic neurons to upregulate their synthesizing capacity and increase the efficiency of dopamine neurotransmission chronically after TBI. PMID- 11496105 TI - Exogenous glutamate induces short and long-term potentiation in the rat medial vestibular nuclei. AB - In rat brain stem slices, high concentrations of exogenous glutamate induce long term potentiation (LTP) of the field potentials evoked in the medial vestibular nuclei (MVN) by vestibular afferent stimulation. At low concentrations, glutamate can also induce short-term potentiation (STP), indicating that LTP and STP are separate events depending on the level of glutamatergic synapse activation. LTP and STP are prevented by blocking NMDA receptors and nitric oxide (NO) synthesis. Conversely, blocking platelet-activating factor (PAF) and group I metabotropic glutamate receptors only prevents the full development of LTP. Moreover, in the presence of blocking agents, glutamate causes transient inhibition, suggesting that when potentiation is impeded, exogenous glutamate can activate presynaptic mechanisms that reduce glutamate release. PMID- 11496106 TI - Enhancing fMRI contrast in awake-behaving primates using intravascular magnetite dextran nanopartieles. AB - Functional MRI in awake-behaving primates is an emerging tool for bridging the gap between human fMRI and neurophysiology information from nonhuman primates. We report the use of magnetite dextran nanoparticles (Feridex) as a blood-pool agent to enhance fMRI contrast-to-noise (CNR) in primate FMRI. The intravascular half life of the magnetite dextran was long compared to lanthanide chelates (T(1/2) = 198 min) with shortened T(2) relaxation observed in blood and cerebral cortex. Greater than 3-fold enhancement in the percentage MR signal change was observed using nanoparticles (13%) compared with conventional BOLD fMRI (4%). The calculated regional cerebral blood volume in macaque primary visual cortex increased 32% with photic stimulation. The increased CNR allows greater flexibility in the design of awake-behaving primate fMRI studies with the potential for improvements in resolution and significantly shortened imaging times. PMID- 11496107 TI - Direct projections from serotonergic neurons in the dorsal and median raphe nuclei of midbrain to the suprachiasmatic nucleus in Tupaia belangeri chinensis. AB - This study investigated the direct serotonergic projections to the suprachiasmatic nucleus (SCN) from the dorsal and median raphe nuclei (DR/MR) of the midbrain in Tupaia belangeri chinensis (TBC) by combined application of retrograde horseradish peroxidase (HRP) tract tracing, immunohistochemistry, and electron microscope techniques. The results provide evidence for the direct projections to the SCN from serotonergic neurons distributed predominantly in the MR (mainly in its lateral portion) and to a lesser degree in the DR (in its ventrolateral portion) more caudally in the midbrain, and the existence of abundant symmetrical and asymmetrical synaptic connections between the serotonergic terminals and the postsynaptic elements in the SCN TBC. The results also revealed that almost all DR neurons projecting to the SCN contained serotonin, whereas about one-half of MR neurons projecting to the SCN were immunoreactive for serotonin. PMID- 11496108 TI - Quantitative measurements of mouse brain thrombin-like and thrombin inhibition activities. AB - Thrombin-like enzymatic activity was measured in mouse brain homogenates and slices by cleavage of a peptide substrate, N-p-Tosyl-Gly-Pro-Arg-7-amido-4 methylcoumarin. The activity was localized mainly to white matter. However, it was not affected by specific thrombin inhibitors, and was found to represent the sum of at least two enzyme activities, a prolyl endopeptidase and an aminopeptidase. By specifically inhibiting this endogenous activity in combination with exogenously added thrombin, mouse brain tissue was shown to express a capacity of thrombin inhibitory activity equivalent to 0.2 mU thrombin/mg brain tissue. The present study offers a simple and reliable method for measuring total thrombin inhibitory activity in brain. PMID- 11496109 TI - Frequency-dependent LTP/LTD in guinea pig Deiters' nucleus. AB - Synaptic plasticity was studied in the lateral vestibular nuclei (LVN) of the guinea pig in vivo. High frequency stimulation (HFS) of increasing or decreasing frequencies was applied to the ipsilateral vestibular nerve. Vestibular field potentials (VFPs) and extracellular single unit activity evoked in the LVN by electrical stimulation of the ipsilateral vestibular nerve, were analyzed before and after the application of different protocols of HFS. Results show that the monosynaptic component of the VFPs undergo long-term potentiation (LTP) with stimulation of 100 Hz applied for 20 s lower frequencies, applied for shorter periods, induce only a transient post-tetanic potentiation. This potentiation, although long lasting, is not permanent since it is susceptible of a reversal or cancellation by opposite patterns of HFS that determine a depression or depotentiation of the previously acquired potentiation. The results demonstrate that the plasticity phenomena that take place at the level of the LVN neurons are not steady but undergo continuous adjustment of their sign and gain depending on the variable flow of vestibular information that reach the nuclei from the labyrinthine receptors. PMID- 11496110 TI - Audiogenic seizure susceptibility in thyroid hormone receptor beta-deficient mice. AB - As early-onset hypothyroidism produces audiogenic seizure susceptibility (AGS) in rodents, the role of TR alpha 1 and TR beta thyroid hormone receptors in AGS was investigated. AGS occurs in mice lacking specifically TR beta (Thrb(tm1/tm1)) and is marked by early onset and persistence, thereby differing from mouse strains where AGS is age-restricted. Thrb(tm1/tm1) mice display AGS whether on a mixed 129/Sv x C57BL/6J or congenic C57BL/6J background. 27% of wild-type mice on the mixed and 0% on the congenic background exhibited AGS. The inability of Thrb(tm1/tm1) mice to downregulate the response to sustained acoustic stimulation may reside in the brain or in the auditory system itself as Thrb(tm1/tm1) mice also display auditory deficits. The AGS phenotype identifies a novel neurological role for TR beta. PMID- 11496111 TI - Co-expression of wild-type and mutant olfactory cyclic nucleotide-gated channels: restoration of the native sensitivity to Ca(2+) and Mg(2+) blockage. AB - In the pore of homomeric cyclic nucleotide-gated (CNG) channels, Ca(2+) and Mg(2+) bind to a set of glutamate residues, which in the bovine olfactory CNG channel are located at position 340. However, native CNG channels from olfactory sensory neurons are composed by the assembly of three different types of subunits, each having a different residue -- glutamate, aspartate or glycine -- at the position corresponding to the binding site for external Ca(2+) and Mg(2+). We co-expressed the wild-type principal alpha subunit with its mutants E340G and E340D in different combinations in Xenopus laevis oocytes, and measured Ca(2+) and Mg(2+) blockage in excised outside-out membrane patches. The comparison between our results and data from native olfactory CNG channels indicates that the presence of all three residues -- glutamate, aspartate and glycine -- in the different subunits, is necessary to restore the sensitivity to external Ca(2+) and Mg(2+) measured in native channels. PMID- 11496112 TI - Event-related TMS over the right posterior parietal cortex induces ipsilateral visuo-spatial interference. AB - The right posterior parietal cortex (PPC) is implicated in visuo-spatial processing, as illustrated by patients with visuo-spatial neglect, but the precise time-course of its contribution is still an open question. In the present study we assessed whether single-pulse transcranial magnetic stimulation (TMS) can interfere with the performance of normal subjects in a standard visuo-spatial task. Participants had to perform a landmark task while TMS was applied over the right PPC, the homologue region in the left hemisphere or the right primary motor cortex. Stimulation was time-locked to the stimulus presentation with a stimulus onset asynchrony (SOA) varying between 50 and 200 ms. Our results indicate that TMS interfered mainly with the visuo-spatial task when applied over the right PPC at an early stage (50 ms post-stimulus). The interference effect of single-pulse TMS in the present visuo-spatial processing is revealed by a processing cost for ipsilateral targets. These results are in agreement with neuropsychological and brain imaging studies showing a right hemispheric dominance in visuo-spatial processing but add crucial information about the time-course of visuo-spatial processing within the right PPC. PMID- 11496113 TI - Reduced corpus callosum, fornix and hippocampus in PDAPP transgenic mouse model of Alzheimer's disease. AB - Persons with Alzheimer's disease (AD) have progressive reductions in the relative sizes of the corpus callosum and hippocampus. Homozygotic PDAPP transgenic mice over-expressing a mutant form of the human amyloid precursor protein have more pronounced reductions in these regions, which are apparent prior to the deposition of amyloid plaques and do not progress with advancing age. The length of the corpus callosum was reduced by two-thirds, the fornix commissure was negligible, and the hippocampal volume was reduced by one-third, suggesting a massive disconnection between the cerebral hemispheres and the hippocampi in PDAPP mice. These findings, which might account for the early, nonprogressive behavioral abnormalities observed in these animals, have implications for the study of AD. PMID- 11496114 TI - Spatial attention triggered by eye gaze increases and speeds up early visual activity. AB - What are the neuronal correlates of reflexive shifts of attention triggered by eye gaze direction? Event related potentials (ERPs) were measured on 14 subjects performing a spatial attention task where eye gaze direction of a face cued the location of a forthcoming target. Subjects were faster in detecting a validly cued target, i.e. one appearing at the location the eye was gazing at, compared to invalidly cued targets, despite the non-predictive value of the eye cues. ERP results showed an enhanced and earlier occipito-parietal P1 and N1 for valid trials, demonstrating the early modulation of visual input by attentional allocation. These findings provide the first evidence that social attention can rapidly modify the processing of visual information in extrastriate cortex. PMID- 11496115 TI - Multidrug resistance-associated protein is involved in the regulation of extracellular levels of phenytoin in the brain. AB - The mechanisms that lead to drug resistance in epilepsy are not known. Recently, overexpression of multidrug transporters, such as multidrug resistance-associated protein (MRP), has been reported in surgically resected epileptogenic human brain tissue and suggested to contribute to the drug resistance of epilepsy. However, it is not known to what extent multidrug transporters such as MRP are involved in transport of antiepileptic drugs. In the present study, we used in vivo microdialysis in rats to study whether the concentration of phenytoin in the extracellular fluid of the cerebral cortex can be enhanced by inhibition of MRP, using the MRP inhibitor probenecid. Local perfusion with probenecid via the microdialysis probe significantly enhanced the extracellular concentration of phenytoin. The data indicate that MRP critically participates in the regulation of extracellular brain concentrations of the major antiepileptic drug phenytoin. PMID- 11496116 TI - Sustained human chemosignal unconsciously alters brain function. AB - The human chemosignal, Delta 4,16-androstadien-3-one modulates psychological state without being consciously discernible as an odor. This study demonstrates that Delta 4,16-androstadien-3-one (androstadienone) alters cerebral glucose utilization both in subcortical regions and in areas of the neocortex not exclusively associated with olfaction. These widely distributed changes are consistent with modulation of an integrated neural network for regulation of emotional and attentional states. This is the first study to demonstrate the effects of a sustained chemosignal on brain metabolism and to show that they are similar to those of long acting chemical substances that affect psychological states. Moreover, this provides the first evidence that a human chemosignal has distributed effects on cortical processes and brain metabolism even when it is not detected consciously. PMID- 11496117 TI - Enhanced frontal cortex activation in rats by convergent amygdaloid and noxious sensory signals. AB - The modulation of frontal cortical EEG activation to noxious somatosensory (tail pressure) and olfactory (acetone) stimulation by the basal amygdala was examined in urethane-anesthetized rats. Mild tail pressure produced no EEG activation, while acetone (sniffed by freely breathing rats or drawn across the olfactory epithelium in tracheotomized rats) produced a moderate suppression of large amplitude synchronized EEG patterns. Concurrent, low-intensity 100 Hz stimulation of the basal amygdala permitted EEG activation to tail pressure to occur, and strongly enhanced olfactory-induced cortical activation. These results indicate that excitation of the basal amygdala potentiates frontal cortical responsiveness to aversive sensory events. This may provide a mechanism to facilitate cortical excitability and processing by amygdaloid neuronal activity. PMID- 11496118 TI - Feeding-related immune responsive brain stem neurons: association with CGRP. AB - Using dual-labeling in situ hybridization histochemistry, the neurotransmitter expression of immune-responsive neurons in the pontine parabrachial nucleus, a major relay for interoceptive information, was investigated. Intravenous injection of bacterial wall lipopolysaccharide resulted in dense c-fos mRNA expression in the external lateral parabrachial nucleus, and a majority of the c fos expressing cells also expressed calcitonin gene-related peptide (CGRP) mRNA. In contrast CGRP-positive cells in the adjoining external medial subnucleus were c-fos negative. Taken together with previous hodological and behavioral studies, these data suggest that CGRPergic parabrachial neurons may mediate lipopolysaccharide-induced anorexia by means of their projection to central nucleus of the amygdala. PMID- 11496119 TI - Up-regulated eNOS protects blood-retinal barrier in the L-arginine treated ischemic rat retina. AB - Using immunoblot analysis and immunocytochemistry, we investigated expression and cellular localization of endothelial nitric oxide synthase (eNOS) and proliferating cell nuclear antigen (PCNA) in the l-arginine treated ischemic rat retina. In parallel, we tested whether the blood-retinal barrier was intact by immunocytochemistry using an antiserum against IgG. In the l-arginine-treated ischemic retina, the magnitude of the increased eNOS was higher, and PCNA was expressed in endothelial cells as well as in neurons in the inner retina during the whole experimental period. Finally, IgG leakage was not detectable in the l arginine-treated ischemic retina. Our results clearly suggest that the increased NO production by eNOS may be essential for the survival of endothelial cells in the rat retina following transient ischemia. PMID- 11496120 TI - The effects of single-trial averaging upon the spatial extent of fMRI activation. AB - We examined effects of trial averaging upon spatial extent, spatial topography, and temporal properties of fMRI activation. Two subjects participated in an event related visual stimulation design. There was an exponential relation between number of trials and spatial extent, such that additional trials identified, on average, a constant proportion of the remaining voxels. At values typical of fMRI experimentation (e.g. 50 trials) only about 50% of eventually active voxels were significant; asymptotic values were approached by 150 trials. The variability of the estimated hemodynamic response decreased with signal averaging, becoming stable across samples of > or = 25 trials. Therefore, group or condition differences may result from differences in voxelwise noise exacerbated by averaging small numbers of trials. PMID- 11496121 TI - Nuclear translocation of extracellular signal-regulated kinases in neuronal excitotoxicity. AB - Subcellular distributions of extracellular signal-kinases (ERK1/2), including their activated form (p-ERK1/2), were investigated in glutamate-induced apoptotic like death in cultured rat cortical neurons by Western immunoblot and immunocytochemistry. During 15 min glutamate exposure, p-ERK1/2 was increased in both cytosol and nuclear extracts, but prominently so in nuclear extracts. Simultaneously, ERK1/2 were mildly decreased in cytosol (to 0.7-fold vs sham control), largely increased in nuclear extracts (to 6.2-fold vs sham control), but not changed in total cell extracts. Immunocytochemistry studies also showed a large increase in nuclear and a mild decrease in cytosol extracts of ERK1/2 at 15 min of exposure. After glutamate exposure, all the above changes reverted simultaneously. The nuclear increase of ERK1/2 was largely prevented by inhibition of ERK1/2 activation, but prolonged by elongation of ERK1/2 activation. These observations suggest that stimulation of glutamate receptors in cortical neurons may incur an activation-dependent transient nuclear translocation of ERK1/2, which might be involved in excitotoxicity through a simultaneous strong elevation of p-ERK1/2 in nucleus. PMID- 11496122 TI - Differential nociceptive responses in mice lacking the alpha(1B) subunit of N type Ca(2+) channels. AB - The role of N-type Ca(2+) channels in nociceptive transmission was examined in genetically engineered mice lacking the alpha(1B) subunit of N-type channels and in their heterozygote and wild-type littermates. In alpha(1B)-deficient mice, N type channel activities in dorsal root ganglion neurons and spinal synaptoneurosomes were eliminated without compensation by other types of voltage dependent Ca(2+) channels. The alpha(1B)-deficient mice showed a diminution in the phase 2 nociceptive responses more extensively than in the phase 1 nociceptive responses of the formalin test. The alpha(1B)-deficient mice exhibited significantly increased thermal nociceptive thresholds in the hot plate test, but failed to increase mechanical nociceptive thresholds in the tail pinch test. These results suggest a crucial role of N-type channels in nociceptive transmission, especially for persistent pain like phase 2 of the formalin test and for nociception induced by thermal stimuli. PMID- 11496123 TI - Intra-supraoptic nucleus sulpiride improves anorexia in tumor-bearing rats. AB - Previous studies suggest that the dopaminergic system in the supraoptic nucleus (SON) is involved not only in the water balance control but also in the food intake regulation. Since we reported that an injection of the D2 receptor antagonist, sulpiride, into specific hypothalamic nuclei (e.g. the LHA, or the VMN) increases food intake in anorectic tumor-bearing rats, as well as in normal rats, we hypothesized that an injection of sulpiride into the SON would also improve cancer anorexia. Sulpiride injection (4 microg/0.5 microl) into bilateral SON of anorectic tumor-bearing male rats significantly improved food intake via increases in both meal size and meal number. These data suggest that pharmacological manipulation of the hypothalamic dopaminergic system is feasible in amelioration of cancer anorexia. PMID- 11496124 TI - Investigating the functional anatomy of empathy and forgiveness. AB - Previous functional brain imaging studies suggest that the ability to infer the intentions and mental states of others (social cognition) is mediated by medial prefrontal cortex. Little is known about the anatomy of empathy and forgiveness. We used functional MRI to detect brain regions engaged by judging others' emotional states and the forgivability of their crimes. Ten volunteers read and made judgements based on social scenarios and a high level baseline task (social reasoning). Both empathic and forgivability judgements activated left superior frontal gyrus, orbitofrontal gyrus and precuneus. Empathic judgements also activated left anterior middle temporal and left inferior frontal gyri, while forgivability judgements activated posterior cingulate gyrus. Empathic and forgivability judgements activate specific regions of the human brain, which we propose contribute to social cohesion. PMID- 11496125 TI - Rewarming eliminates the protective effect of cooling against delayed neuronal death. AB - Mild intra-ischemic hypothermia provides neuroprotection against delayed neuronal death in the hippocampal CA1. It has recently been reported that reduction in the metabolic rate of arachidonic acid (AA) liberated during ischemia might contribute to this neuroprotection. To examine whether rewarming during the early period of recirculation accelerates AA consumption and eliminates the neuroprotection, we measured the levels of AA in the hippocampus after various recirculation times under normothermia and hypothermia with or without rewarming. The tendency for AA to disappear was significantly different between each pair of groups. Histological examination 7 days after ischemia revealed no protection in the rewarmed group. These results suggest that neuronal injury during rewarming after hypothermia may be attributed to the rate of AA metabolism. PMID- 11496126 TI - Measuring temporal resolution in infants using mismatch negativity. AB - We show that the mismatch negativity (MMN) component of the event-related potential can be used to measure auditory temporal resolution in human infants. Infrequent stimuli with silent gaps of 4, 8, or 12 ms modulated the P2 component, generated MMN, and produced a P3a-like positivity. The data indicate that within channel gap detection thresholds at 6 months are essentially at adult levels under conditions of little adaptation. Since MMN is elicited without attention and does not require a behavioural response, it can be measured similarly across the lifespan. We are now in a position to study the development of cross-channel temporal resolution and adaptation effects in infancy, and to examine how these abilities in infancy relate to later language acquisition. PMID- 11496127 TI - Recruitment of activated microglia cells in the spinal cord of mice by ALS IgG. AB - Mice were injected i.p. with IgG samples of different patients to test whether IgG from amyotrophic lateral sclerosis (ALS) can initiate an immune/inflammatory reaction targeting motor neurons. All IgG samples of five ALS patients and none of the disease controls recruited activated microglia cells in the ventral horn of the spinal cord. CD3 lymphocytes were not accumulated in the same tissue. Similar reaction was evoked by injection of IgG from guinea pigs with experimental autoimmune gray matter disease (EAGMD) induced by immunization with the homogenate of the ventral horn of bovine spinal cord. The results indicate that ALS IgG and anti-motoneuron IgG induce microglia reaction targeting motor neurons without initiating T cell response in the recipient mice. PMID- 11496128 TI - Inhibition of myopia development in chicks using himbacine: a role for M(4) receptors? AB - The success of the M(1)-selective muscarinic antagonist pirenzepine in preventing myopia development in animal models implicates a role for the M(1) receptor. However, the relatively high dose of pirenzepine required may indicate that the drug acts through another receptor subtype. This study examined whether the M(4) selective antagonist, himbacine, could also prevent myopia. Daily intravitreal injections of himbacine inhibited the inducement of myopia in chick eyes in a dose- dependent manner. Doses < or = 200 microg caused no significant inhibition of induced myopia compared to controls (-13.7 +/- 2.3 vs -16.2 +/- 0.9D, ANOVA p = 0.37), whilst a dose of 800 microg almost completely inhibited the induced myopia (-2.4 +/- 2.0, p < 0.01). Findings demonstrate himbacine is effective at preventing the development of myopia in chick and implicates a role for the M4 receptor. PMID- 11496129 TI - Muscarinic M2 receptors inhibit heat-induced CGRP release from isolated rat skin. AB - The action of cholinergic agonists on modulating basal and heat-induced CGRP release was investigated in isolated rat skin. Nicotine (10(-6), 10(-5) and 10( 4) M) induced a bimodal increase of CGRP release, that was significant for the two larger concentrations (by 113 and 36%, respectively). On the contrary, muscarine (10(-4) M) and arecaidine (10(-5) M) significantly decreased the basal CGRP release (by 16 and 23%, respectively). The substantial increase of CGRP release evoked by noxious heat (47 degrees C) remained unaltered upon co application of nicotine, but was diminished by 35% upon muscarine. Arecaidine was more effective in this respect causing significant dose-dependent depressions by 30% (at 10(-6) M) and by 60% (at 10(-5) M). These data support a role of muscarinic M2 receptors in nociceptor desensitization. PMID- 11496130 TI - Expression of Id proteins increases in astrocytes in the hippocampus of epileptic rats. AB - Reactive gliosis is a prominent morphological feature of temporal lobe epilepsy. The molecular mechanisms underlying glial cell activation remain unclear. We examined expression of Id1-3 protein, a family of helix--loop--helix proteins involved in the regulation of cell proliferation and differentiation, in glial cells after electrically induced status epilepticus (SE) in the rat. In control hippocampus, Id3 was weakly expressed in astrocytes, while Id1-2 were below detection level. After SE, Id1-3 protein expression increased markedly in reactive astrocytes within 1 day and this persisted up to 3 weeks after SE. Three months after SE when rats experience spontaneous seizures, Id expression had returned to control levels. These results support a role of the Id gene family in regulating astrocyte reactivity in epileptic tissue. PMID- 11496131 TI - Brain lateralization for mismatch response to across- and within-category change of vowels. AB - Differences in hemispheric predominance between across- and within-category change perception of vowels were assessed using a whole-head magnetoencephalography. The magnetic mismatch responses (MMNm) to pure-tone and vowel within-category changes were significantly predominant in the right hemisphere; on the other hand, vowel across-category MMNm did not differ in power between hemispheres. The results suggest that both hemispheres are symmetrically activated in the preattentive across-category change perception of vowels, while the within-category change of a vowel is analyzed as the change in physical features of the stimuli, thus predominantly activating the right hemisphere. Thus, the relative contribution of the left auditory cortex in the preattentive speech processing may occur only at the level of perception of the vowel across category change. PMID- 11496132 TI - Alteration of rCBF in Alzheimer's disease patients with delusions of theft. AB - We investigated the neural substrate of the delusion of theft in patients with Alzheimer's disease (AD). Nine AD patients with only one type of delusion (delusions of theft) and nine age, cognitive function-matched AD patients without any type of delusions were selected from 334 consecutive outpatients of Ehime University Hospital. All subjects underwent (99m)Tc-HMPAO SPECT scanning, and SPECT images were analyzed by Statistical Parametric Mapping (SPM). AD patients with delusions of theft showed significant hypoperfusion in the right medial posterior parietal region compared to patients without delusions. Our data suggest that attention impairment or lack of awareness of illness caused by right parietal dysfunction might play a role in producing the delusion of theft. PMID- 11496133 TI - Cell genesis in the hypothalamus is associated to the sexual phase of a hermaphrodite teleost. AB - The present study aimed to define sex differences in the genesis of hypothalamic and telencephalic cells in the adult brain of male, female and under sex reversal teleosts (Sparus aurata). Application of 5-bromo-2-deoxyuridine immunocytochemistry and quantification of the newborn cells using the disector method, determined a statistically significant difference in the dorsal hypothalamic ventricular area between sexual phases. Female brain exhibited a higher number of newborn cells, suggesting that estrogens possibly influence higher mitotic activity. In contrast, paraventricular organ and ventral nucleus of the ventral telencephalon did not show any significant changes, while cell proliferation differences in the preoptic area were correlated with age. The sex specific cell proliferation pattern in the hypothalamus adds important information on the cellular mechanisms that underlie sex change in a protandrous hermaphrodite teleost. PMID- 11496134 TI - Multiple tactile maps in the human cerebellum. AB - Functional imaging studies of the cerebellum have mostly investigated motor performance or have been limited to the anterior lobe and therefore the somatosensory representations in the human cerebellum have not been fully demonstrated. We used fMRI of the entire cerebellum during tactile stimulation of the hand and foot in six normal subjects. Our results demonstrate that the tactile projections to the cerebellum in humans are represented in both the anterior and posterior lobes. in agreement with previous functional imaging studies, our results show a large-scale, between-limb somatotopy comparable to that shown in early animal studies. PMID- 11496135 TI - Interhemispheric phase synchrony and amplitude correlation of spontaneous beta oscillations in human subjects: a magnetoencephalographic study. AB - Interhemispheric phase synchrony and amplitude correlation of beta oscillations were studied with MEG in a resting condition. The left and right hemisphere beta oscillations exhibited phase-locking with a phase-lag near zero degrees. The index of synchronization was strongest when these oscillations had large amplitude. Functionally, we interpret the phase synchrony on the basis of bilaterality of movement organization. A positive interhemispheric correlation was also found for the amplitude of spontaneous beta oscillations over long time intervals (> 1 s). The low-frequency correlation of spontaneous rhythmic activity may be the source of the low-frequency correlations of the hemodynamic responses in homologous areas that have been reported previously and have been interpreted as functional connectivity between these areas. PMID- 11496136 TI - beta amyloid-induced disruption of ionic balance: studies on the isolated frog labyrinth. AB - The beta-amyloid peptide (A beta) is a key player in the pathogenesis of Alzheimer's disease. Although its mechanisms of action are not fully elucidated, a disruption of ionic homeostasis has been suggested, and A beta aggregation in fibrils seems correlated to its toxic potential. In the present work, we studied the effects of different A beta fragments on the activity of frog ampullar nerve fibers. Our results show that A beta fragments are able to reduce ampullar nerve responses, with a potency correlated to their fibrillogenic capability. This study may have clinical implications, since vestibular problems are often reported in Alzheimer patients, and provide a model for the dissection of A beta effects in a simple multicomponent system. PMID- 11496137 TI - Medullary A1 noradrenergic neurones may mediate oxytocin release after noxious stimuli. AB - Noxious stimuli facilitate oxytocin release from the pituitary. Oxytocin cells receive excitatory synaptic inputs from the noradrenergic neurones located in the medulla oblongata. Oxytocin release after noxious stimuli is blocked by noradrenaline depletion in the brain. Here, we examined effects of noxious stimuli upon noradrenaline release within the supraoptic nucleus. Electric footshocks or mustard oil application to the foot pad facilitated noradrenaline release in the nucleus. Noradrenaline release after noxious stimuli was impaired by microinjections with a GABA(A) receptor agonist, muscimol, or an alpha 2 adrenoceptor agonist, clonidine, into the A1 noradrenergic cell regions. From these and reported data, we conclude that the medullary A1 noradrenergic neurones contribute, at least in part, to oxytocin release from the pituitary after noxious stimuli. PMID- 11496138 TI - Alternating dominance of NMDA and AMPA for learning and recall: a computer model. AB - Physiological studies reveal a dichotomy in biological Hebbian learning: NMDA receptors are utilized for induction of long term potentiation (LTP) whereas AMPA is used for LTP expression. We propose that this dichotomy would have functional value: preventing previously stored memories from interfering with the storage of new memories. A previous hypothesis reduces this interference by temporarily reducing associative weights during learning. Complementary to this model, we propose a dual transmission algorithm in which one set of synaptic weights are used primarily for learning and another primarily for recall. This algorithm shows good performance in a simple neural network model. Biologically, the model could be mediated by a cholinergic switch from dominance of learning-insensitive NMDA receptors to dominance of learning-modifiable AMPA receptors. PMID- 11496139 TI - The modulatory effect of estrogen on the neuronal activity in the barrel cortex of the rat. An electrophysiological study. AB - In acute experiments, the effects of iontophoretically applied 17 beta-estradiol hemisuccinate on the activity of the primary somatosensory cortical neurons were studied in ovariectomized rats by extracellular single-unit recording. 17 beta Estradiol increased both the spontaneous and the vibrissa deflection-evoked responses, with an average latency of 24 min. It is suggested that this relatively long latency of the 17 beta-estradiol effect is based not so much on membrane mechanisms as on genomic mechanisms. PMID- 11496140 TI - Astrocytes induce manganese superoxide dismutase in brain capillary endothelial cells. AB - Astrocytes induce blood-brain barrier (BBB) properties in brain endothelial cells (EC)*O(2)*, generated in blood and EC, opens the BBB. Hence, high activity of superoxide dismutase (SOD) is a prerequisite for normal BBB function. Therefore, the influence of rat astrocytes on the expression of manganese (Mn)SOD in rat EC was investigated in two coculture models of the BBB, allowing either exchange of soluble factors or additionally cellular contacts. Activity, protein content and mRNA expression of endothelial MnSOD were significantly increased in both coculture models in comparison to monoculture by soluble astrocytic factors, such as cytokines. High activity of endothelial MnSOD may be considered as a further essential property of the BBB, which is induced and maintained by astrocytes. PMID- 11496141 TI - Imagining rotation by endogenous versus exogenous forces: distinct neural mechanisms. AB - Previous neuroimaging studies of mental image transformations have sometimes implicated motor processes and sometimes not. In this study, prior to neuroimaging the subjects either viewed an electric motor rotating an angular object, or they rotated the object manually. Following this, they performed the identical mental rotation task in which they compared members of pairs of such figures, but were asked to imagine the figures rotating as they had just seen the model rotate. When results from the two rotation conditions were directly compared, motor cortex (including area M1) was found to be activated only when subjects imagined the rotations as a consequence of manual activity. Thus, there are at least two, qualitatively distinct, ways to imagine objects rotating in images, and these different strategies can be adopted voluntarily. PMID- 11496142 TI - Responses of rat dorsal column neurons to pancreatic nociceptive stimulation. AB - Our previous study has shown that the postsynaptic dorsal column pathway is involved in transmission of pancreatic nociceptive information from the spinal cord to the thalamus. The aim of the present study was to characterize the neuronal responses in the dorsal column nuclei of the rat to pancreatic nociceptive stimulation. Brief application of bradykinin (10 microg/ml, 20s) to the surface of the pancreas resulted in a significantly increased firing rate in approximately 10% of neurons recorded. In conclusion, pancreatic nociceptive information relayed to the dorsal column nuclei is transmitted from the spinal cord through the dorsal columns. PMID- 11496143 TI - Delayed application of IGF-I and GDNF can rescue already injured postnatal motor neurons. AB - IGF-I, GDNF, and other neurotrophic factors, when applied at the time of injury, can protect postnatal motor neurons from slow glutamate injury in organotypic spinal cord. However, in human spinal cord diseases, motor neuron injury is already established when treatment could begin. We tested whether neurotrophic factors can protect already-injured motor neurons, and whether combinations of factors can further lengthen the therapeutic time window. Our data show that during a 7--8 week process of slow neurodegeneration either IGF-I or GDNF treatment, though delayed up to 4 weeks, still allowed substantial rescue of already injured motor neurons. However, the combination of both factors additively provided better neuroprotection than either factor alone, even after a 4-week delay. This proof of principle is relevant to the potential of IGF-I and GDNF as therapy for acquired disorders affecting motor neurons. PMID- 11496144 TI - Chloride accumulation and depletion during GABA(A) receptor activation in neocortex. AB - The response of neocortical neurons to GABA is strongly influenced by the intracellular chloride concentration. We tested the hypothesis that activation of GABA(A) receptors can result in either depletion or accumulation of intracellular chloride. The measured reversal potentials of currents evoked by exogenously applied or synaptically released GABA were not significantly different. During GABA responses, voltage steps to the reversal potential revealed prominent tail like currents. The polarity of these currents was opposite to that of the GABA evoked currents, consistent with either accumulation or depletion of intracellular chloride. These results demonstrate that currents evoked by exogenously applied and synaptically released GABA share similar ionic dependencies. Current fluxes during GABA(A) receptor activation can be sufficiently large to change the intracellular chloride concentration. PMID- 11496145 TI - Sex differences in amygdala activation during the perception of facial affect. AB - The cognitive and affective systems of the cerebral cortex are often more lateralized in males than females, but it is unclear whether these differences extend to subcortical systems. We used fMRI to examine sex differences in lateralized amygdala activity during happy and fearful face perception. Amygdala activation differed for men and women depending on the valence of the expression. Overall, males were more lateralized than females, but the direction differed between valence conditions. Happy faces produced greater right than left amygdala activation for males but not females. Both sexes showed greater left amygdala activation for fearful faces. These findings suggest that the lateralization of affective function may extend beyond the cortex to subcortical regions such as the amygdala. PMID- 11496146 TI - Sub-second changes in accumbal dopamine during sexual behavior in male rats. AB - Transient (200--900 ms), high concentrations (200--500 nM) of dopamine, measured using fast-scan cyclic voltammetry, occurred in the nucleus accumbens core of male rats at the presentation of a receptive female. Additional dopamine signals were observed during subsequent approach behavior. Background-subtracted cyclic voltammograms of the naturally-evoked signals matched those of electrically evoked dopamine measured at the same recording sites. Administration of nomifensine amplified natural and evoked dopamine release, and increased the frequency of detectable signals. While gradual changes in dopamine concentration during sexual behavior have been well established, these findings dramatically improve the time resolution. The observed dopamine transients, probably resulting from neuronal burst firing, represent the first direct correlation of dopamine with sexual behavior on a sub-second time scale. PMID- 11496147 TI - Parvalbumin-containing interneurons do not innervate granule cells in the olfactory bulb. AB - Combining pre-embedding parvalbumin immunostaining and post-embedding immunogold detection of GABA in the olfactory bulb, we investigated whether the parvalbumin containing GABAergic interneurons of the external plexiform layer exclusively innervate principal cells, or whether they also establish inhibitory synapses upon GABAergic local neurons such as granule cells. Our results demonstrate that the parvalbumin-containing cells do not contact GABAergic interneurons in the neuropil of the external plexiform layer. On the contrary, their postsynaptic elements were always non-GABAergic principal cells. Although classically it has been accepted that the interneurons of the external plexiform layer could exert a disinhibitory action upon principal cells, via inhibition of GABAergic granule cells, we conclude that they exert a feedback inhibitory action directly and exclusively upon principal cells. PMID- 11496148 TI - Cross-modal integration of auditory and visual motion signals. AB - Real-world moving objects are usually defined by correlated information in multiple sensory modalities such as vision and hearing. The aim of our study was to assess whether simultaneous auditory supra-threshold motion introduces a bias or affects the sensitivity in a visual motion detection task. We demonstrate a bias in the perceived direction of visual motion that is consistent with the direction of the auditory motion (audio-visual motion capture). This bias effect is robust and occurs even if the auditory and visual motion signals come from different locations or move at different speeds. We also show that visual motion detection thresholds are higher for consistent auditory motion than for inconsistent motion, provided the stimuli move at the same speed and are co localised. PMID- 11496149 TI - Identification of amyloid-beta binding sites using an antisense peptide approach. AB - The amyloid-beta (A beta) peptide is a cytotoxic peptide implicated in the pathology of Alzheimer's disease (AD). Catalase and the endoplasmic reticulum A beta binding dehydrogenase (ERAB) are both inhibited by characterized fragments of the A beta peptide. In order to target such proteins it is essential to determine which components of these enzymes interact with A beta. This study reports the use of antisense peptide methodology to identify specific A beta binding domains. Synthetic peptides corresponding to the regions of catalase and ERAB identified showed specific binding to A beta and also prevented A beta cytotoxicity. Antisense peptide methodology has identified A beta recognition sequences and may also be applied to the identification of novel A beta protein interactions to identify targets for use in the treatment of AD. PMID- 11496150 TI - Microglial signaling by amyloid beta protein through mitogen-activated protein kinase mediating phosphorylation of MARCKS. AB - Myristoylated alanine-rich C kinase substrate (MARCKS), an acidic protein associated with cell motility and phagocytosis, is activated upon phosphorylation by protein kinase C (PKC) and proline-directed protein kinases. In Alzheimer disease (AD), activated microglia expressing MARCKS migrates around senile plaques. We reported that amyloid beta protein (A beta), a major component of senile plaques, activated MARCKS through a tyrosine kinase and PKC-delta. We have now identified another A beta signaling pathway through a mitogen-activated protein kinase (MAPK) involved in the phosphorylation of MARCKS and analysed cross-talk between PKC and MAPK pathways in primary cultured rat microglia. A selective inhibitor for MAPK kinase, PD098059, significantly inhibited the phosphorylation of MARCKS induced by A beta. Extracellulary regulated kinases, the activities of which were induced by A beta, directly phosphorylated a recombinant MARCKS in vitro. The MAPK pathway was sensitive to wortmannin, but not to a PKC inhibitor or to tyrosine kinase inhibitors. The activation of PKC by A beta was not sensitive to wortmannin. Our findings suggest involvement of the MAPK pathway through phosphoinositol 3-kinase in the phosphorylation of MARCKS in rat cultured microglia, an event may be associated with mechanisms activating microglia in AD. PMID- 11496151 TI - Auditory evoked magnetic fields in adults with fragile X syndrome. AB - Hyper-reactivity and anxiety to sensory stimuli have been described in patients with fragile X syndrome (FXS), and may be related to abnormal processing in afferent sensory pathways. We used magnetoencephalography (MEG) to measure auditory responses to pure tones in 11 adults with FXS and 11 non-FXS subjects. The amplitude for the N100m auditory evoked field component was significantly higher for patients with FXS than for subjects. FXS subjects also had less lateralized N100m anterior-posterior dipole locations. These data may suggest that more neurons are activated by acoustic stimuli in FXS, consistent with subjective experience of increased stimulus intensity. Anomalous cerebral lateralization may suggest an early critical window for effects on neocortical development of the fragile X mental retardation protein (FMRP) produced by the FMR1 gene in individuals with FXS. PMID- 11496152 TI - Intermuscular coherence in Parkinson's disease: relationship to bradykinesia. AB - We hypothesised that bradykinesia may be partly due to the failure of the corticomuscular system to engage in high frequency oscillatory activity in Parkinson's disease (PD). In healthy subjects such oscillations are evident in coherence between active muscles at 15--30 Hz. We therefore investigated the effects of therapeutic stimulation of the basal ganglia on this coherence and related it to changes in bradykinesia in the contralateral arm. Increases in coherence at 15--30 Hz and improvements in bradykinesia upon stimulation were correlated (r = 0.564, p < 0.001). This suggests that the basal ganglia modulate oscillatory activity in the corticomuscular system and that impairment of the motor system's ability to engage in synchronised oscillations at high frequency may contribute to bradykinesia in PD. PMID- 11496153 TI - Cerebral mechanisms underlying orienting of attention towards auditory frequency changes. AB - Brain mechanisms underlying detection of auditory frequency changes were studied with event-related potentials (ERPs) in 14 human subjects discriminating visual stimuli. Scalp-current density mapping revealed bilateral components of mismatch negativity (MMN) in frontal and auditory cortices. Deviance-related activations in frontal and temporal cortex began to be significant at 94 ms and 154 ms in the right hemisphere, and at 128 ms and 132 ms in the left hemisphere. The magnitude of MMN-neuroelectric currents from the left temporal cortex correlated significantly (r = -0.56, p < 0.05) with distraction caused by MMN-eliciting deviant tones. These results suggest a complex cerebral circuitry involved in frequency change detection and strongly support the role of this circuitry in driving attention involuntarily towards potentially relevant frequency changes in the acoustic environment. PMID- 11496154 TI - Killing of prion-damaged neurones by microglia. AB - The loss of neurones that occurs in the transmissible spongiform encephalopathies, or prion diseases, can be reproduced in vitro by incubating neuronal cultures with either peptides derived from the prion protein or with partially purified prion preparations. In the present studies, the extent of neuronal loss on exposure to these prions or prion peptides was increased by the addition of microglia, a process that was dependent upon the number of microglia added, the concentration of prions/peptides present and the degree of fibrillarity of the prion peptides. Microglia also killed scrapie-infected neuroblastoma cells expressing infectious PrP(SC). Microglia secreted low amounts of interleukin (IL)-6 when incubated with peptides alone but up to 10 times as much IL-6 when incubated with peptide-treated neurones, suggesting that microglia recognise peptide-induced changes in neurones. PMID- 11496155 TI - Addiction changes orbitofrontal gyrus function: involvement in response inhibition. AB - We used the Stroop task as a measure of the ability to inhibit a prepotent response tendency and examined its association with relative glucose metabolism in selected prefrontal brain regions in cocaine addicts, alcoholics, and controls (17 per group). Results revealed that for the substance abusers, higher orbitofrontal gyrus (OFG) activation was associated with lower conflict (higher score; r = 0.32, p < 0.05). For the controls, higher OFG activation was associated with higher conflict (lower score; r = -0.42, p < 0.05). Thus, at baseline, increased relative activation of the OFG is associated with worse performance in controls and better performance in substance abusers on the Stroop task, suggesting reversal of the role of the OFG as a function of addiction. PMID- 11496156 TI - Visual imagery without visual experience: evidence from congenitally totally blind people. AB - We explored the ability of congenitally totally blind people (who were contrasted with age-, sex- and education matched blindfolded sighted subjects) to perform tasks which are mediated by visual mental imagery in sighted people. In the first (pictorial) task, subjects had to mentally compare the shape of the outline of three named objects and to indicate the odd-one-out. In the second (spatial) task the participants were asked to memorise the position of a target cube in two- and three-dimensional matrices, based on a sequence of spatially based imagery operations. In addition, during half of the trials of both imagery tasks subjects were required to perform a concurrent finger tapping task, to investigate whether the blind subjects would be more dependent on spatial processing. Although blind participants made significantly more errors than sighted participants, they were well able to perform the spatial imagery task as well as the pictorial imagery task. Interference from the concurrent tapping task affected both groups to the same extent. Our results shed new light on the question whether early visual experience is necessary for performance on visual imagery tasks, and strongly suggest that vision and haptics may share common representations. PMID- 11496157 TI - Timing of right parietal and frontal cortex activity in visuo-spatial perception: a TMS study in normal individuals. AB - In a recent study we showed that repetitive transcranial magnetic stimulation (rTMS) with train duration of 400 ms over right frontal and right posterior parietal cortices gives rise to transitory contralateral visuo-spatial neglect in normal subjects. In the present experiment we investigated whether using single pulse TMS it is possible to obtain information about the timing of cortical activity related to spatial cognition. Nine healthy subjects performed in baseline condition and during TMS a tachistoscopic task, requiring a forced choice estimation of the length of the two segments of prebisected horizontal lines. Single-pulse TMS was triggered at various time intervals (150 ms, 225 ms, 300 ms) after visual stimulus onset with a focal coil over P6 and F4 (according to 10/20 EEG system). Relative transitory rightward bias was observed only when parietal TMS was delivered 150 ms after visual stimulus presentation. Frontal stimulation induced no effect on visuo-spatial perception with the time intervals explored. PMID- 11496158 TI - Caspr1/Paranodin/Neurexin IV is most likely not a common disease-causing gene for inherited peripheral neuropathies. AB - Contactin associated protein 1 (Caspr1/Paranodin/Neurexin IV) is an axonal transmembrane molecule mainly localised at the paranodal junction. Since molecular alterations in septate-like junctions at the paranodes might have important consequences for the function of the nerve fiber, we considered that Caspr1 could be involved in the pathogenesis of inherited peripheral neuropathies. In this study, we physically mapped the Caspr1 gene on chromosome 17q21.1 and determined its genomic structure. We performed a mutation analysis of the Caspr1 gene in a cohort of 64 unrelated patients afflicted with distinct inherited peripheral neuropathies. Since no disease causing mutations were found, we suggest that Caspr1 is probably not a common cause of inherited peripheral neuropathies. PMID- 11496159 TI - A guest editorial: dialog between basic and clinical science: relaxin as a possible cause of symphyseal separation. PMID- 11496160 TI - Fibroids and infertility: a systematic review of the evidence. AB - A systematic literature review was performed to determine whether leiomyomata are associated with decreased fertility rates, and whether surgical removal increases fertility rates postoperatively. Meta-analysis was conducted when multiple studies addressed a single issue and were sufficiently homogeneous. Data were analyzed for effect of any fibroid upon fertility, as well as specific fibroid location. Results of studies comparing women with infertility and fibroids versus infertile controls showed widely disparate results. Subgroup analysis failed to indicate any effect on fertility of fibroids that did not have a submucous component. Conversely, women with submucous myomas demonstrated lower pregnancy rates (RR 0.30; 95% confidence interval [CI] 0.13--0.70) and implantation rates (RR 0.28; 95% CI 0.10--0.72) than infertile controls. Results of surgical intervention were similar. When all fibroid locations were considered together, myomectomy results were again widely disparate. However, when women with submucous myomas were considered separately, pregnancy was increased after myomectomy compared with infertile controls (RR 1.72; 95% CI 1.13--2.58) and delivery rates were now equivalent to infertile women without fibroids (RR 0.98; 95% CI 0.45--2.41). The current data suggest that only those fibroids with a submucosal or an intracavitary component are associated with decreased reproductive outcomes, and that hysteroscopic myomectomy may be of benefit. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians LEARNING OBJECTIVES: After completion of this article, the reader will be able to summarize the data on the role of fibroids and infertility, describe the effect of myomectomy on fertility, and list the methods used to evaluate intracavitary fibroids. PMID- 11496161 TI - Osteoporosis in children and adolescent girls: case report of idiopathic juvenile osteoporosis and review of the literature. AB - The diagnosis and treatment of osteoporosis is an important aspect of gynecologic training and practice. Idiopathic juvenile osteoporosis (IJO) is a rare disease of children and adolescents that resolves after the onset of puberty. A case report is presented and current methods of diagnosis and treatment of IJO are discussed as well as the differential diagnosis. A MEDLINE search was performed of the following terms: idiopathic juvenile osteoporosis, pediatric osteoporosis, adolescent osteoporosis, bisphosphonates pediatric adolescent, and pregnancy osteoporosis, and references from bibliographies of selected papers were used as well. All papers in English, French, and German are considered in this review. There were 114 papers selected as relevant to the topic. Data relevant to the diagnosis, pathogenesis, methods of imaging, laboratory evaluation, differential diagnosis, and treatment of IJO are presented. IJO is a diagnosis of exclusion in the pediatric and adolescent patient with osteoporosis. Although bone density gradually improves after the onset of puberty, treatment of currently affected children and adolescents involves activity restriction, calcium, vitamin D, and bisphosphonate therapy. Future reproductive concerns are discussed and areas requiring additional study are reviewed. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians LEARNING OBJECTIVES: After completion of this article, the reader will be able to describe the condition idiopathic juvenile osteoporosis, compare the clinical features of this condition to other similar conditions, outline the diagnostic workup of a child with this condition, and list the potential therapeutic options for a patient with idiopathic juvenile osteoporosis. PMID- 11496162 TI - Practice trends in outpatient obstetrics and gynecology: findings of the Collaborative Ambulatory Research Network, 1995--2000. AB - Historically, obstetrics and gynecology has been a medical/surgical specialty focusing on women's health and reproductive concerns during the childbearing years. Newer responsibilities-for example, in primary care, gerontology, and genetics-require Ob-Gyns to draw upon a base of medical knowledge that traditionally was not considered germane to their practices. Ob-Gyns are increasingly providing more primary care services to their patients; consequently, the field has expanded considerably. The Collaborative Ambulatory Research Network (CARN) was created in 1990 as a vehicle for investigating issues pertinent to women's health and to the practice of obstetrics and gynecology in the outpatient setting. This article summarizes the findings of CARN studies from 1995 to 2000, covering a range of topics related to women's health across the life cycle, including, but not limited to perinatal care. Topics include nutrition, infectious disease, hormone replacement therapy, psychosocial issues, and genetic testing in obstetric and gynecologic practice. Each study produced a picture of current practice patterns and knowledge of the physicians surveyed. Findings on knowledge, attitudes, and practices varied widely. Overall, Ob-Gyns were knowledgeable and consistent in more traditional areas of practice. Conversely, inconsistencies were observed in newer areas. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians LEARNING OBJECTIVES: After completion of this article, the reader will be able to define what the Collaborative Ambulatory Research Network (CARN) involves, describe how CARN obtains its data, and summarize some of the findings of CARN from the years 1995 to 2000. PMID- 11496163 TI - Temporary arteriovenous fistula and microsurgical free tissue transfer for reconstruction of complex defects. PMID- 11496164 TI - Morphometry of the orbital region: a soft-tissue study from adolescence to mid adulthood. AB - The objective of this study was to gain information about normal sex-related linear and angular dimensions of the orbital region; left-right symmetry; and growth changes between adolescence and mid-adulthood. The three-dimensional coordinates of several soft-tissue landmarks on the orbits and face were obtained by an electromagnetic digitizer in 40 male and 33 female adolescents aged 12 to 15 years, 73 female and 89 male young adults aged 19 to 30 years, and 41 male and 38 female adults aged 31 to 56 years. From the landmarks-binocular and intercanthal widths; paired height and inclination of the orbit relative to both the true horizontal (head in natural head position) and Frankfurt plane; length and inclination of the eye fissure; and the orbital height to eye fissure length ratio were calculated and averaged for age and sex. Comparisons were performed by factorial analysis of variance. Both the linear dimensions and the angular values were significantly larger in male subjects than in female subjects of corresponding age (p < 0.05). A significant effect of age was found (p < 0.05): while the linear distances and the orbital height-to-length ratio were greater in older people of the same sex, the inclination of the eye fissure decreased as a function of age. Both orbital inclinations were larger in the young adult group than in the adolescent and middle-aged groups. On average, the paired measurements were symmetrical, with similar values within each sex and age group. Data collected in this investigation could serve as a database for the quantitative description of human orbital morphology during normal adolescent and adult growth. PMID- 11496166 TI - Plasticity of the endocranial base in nonsyndromic craniosynostosis. AB - Limited in vivo data exist on the dysmorphology of the cranial base in nonsyndromic craniosynostosis. Few studies have documented the effect of calvarial surgery for synostosis on endocranial morphology. Previous work has suggested that the dysmorphology of the endocranial base is diagnostically specific for metopic, sagittal, and unicoronal sutures. The purpose of this study was to further evaluate the endocranial base in infants with nonsyndromic craniosynostosis by testing the hypothesis that the dysmorphology is, to some degree, a secondary deformation rather than a primary malformation. Three questions were addressed: (1) Can individuals reliably identify affected suture specific endocranial-base morphology using standard templates? (2) Does calvarial surgery in infancy for craniosynostosis affect the perception of endocranial-base morphology? and (3) Does calvarial surgery in infancy for nonsyndromic craniosynostosis normalize the endocranial base?In this study, three-dimensional volumetric reconstructions from archived computed tomography digital data were processed using the ANALYZE imaging software. Dysmorphology was assessed by nine independent, blinded skilled observers who reviewed two separate sets of images of endocranial bases. Both sets contained images from the same patients: one set contained preoperative images, and the other contained images of the endocranial base 1 year after calvarial surgery. Observers were asked to sort each set into four suture-specific diagnostic groups: normal, unicoronal, metopic, and sagittal. Each set contained 10 patients with unicoronal synostosis, 10 with metopic synostosis, 10 with sagittal synostosis, and four normal patients. Seventy-eight percent of the total number of preoperative images were correctly sorted into the suture-specific diagnostic group, whereas only 55 percent of the total number of postoperative images were correctly matched. With regard to the individual sutures, the results were as follows (data are presented as preoperative accuracy versus postoperative accuracy): metopic, 76 percent versus 44 percent; sagittal, 58 percent versus 34 percent; unicoronal, 100 percent versus 79 percent; and normal, 83 percent versus 72 percent. Although 36 of 306 total images per group (12 percent) actually represented normal patients, the observers called 72 of 306 normal (24 percent) in the preoperative set versus 110 of 306 normal (36 percent) in the postoperative set. In conclusion, (1) the endocranial dysmorphology of nonsyndromic craniosynostosis is recognizably specific to the affected suture; (2) calvarial surgery for nonsyndromic craniosynostosis normalizes the endocranial base qualitatively with regard to the diminished ability of raters to identify the primary pathology; and (3) the documented postoperative changes in endocranial base morphology after calvarial surgery for nonsyndromic craniosynostosis in infancy indicates that a major component of that dysmorphology is a secondary deformity rather than a primary malformation. PMID- 11496167 TI - Mandibular distraction osteogenesis in very young patients to correct airway obstruction. AB - The purpose of this study was to measure changes in the airway cross-sectional area of pediatric patients with micrognathia and obstructive airway symptoms after treatment by mandibular distraction. The measurements obtained were correlated with the clinical outcomes.Ten patients, ranging in age from 3 months to 8 years, underwent measurement and distraction. Eight patients were under 30 months of age. Six were diagnosed with Pierre Robin sequence, two with Treacher Collins syndrome, and two with Nager syndrome. All patients had retrognathia of greater than 8 mm and obstructive airway symptoms while awake that had resulted in tracheostomy (3), repeated apnea monitor triggering (5), or abnormal sleep study (2). Cephalometric analysis was performed pretreatment and posttreatment by distraction. The effective airway space was defined with the following boundaries: a horizontal line from the tip of the odontoid to the velum, the uvula tip to the tongue base along the shortest line, the tongue base down to the base of the epiglottis, and the horizontal line to the posterior pharynx. These lines were traced for each cephalogram, the outline was digitized, and the area was calculated by computer. An analysis of the square area change was done by paired t test. The range of distraction was 8 to 22 mm; the mean effective airway increase was 67.5 percent, with a range of 26 to 120 percent. Measurable airway increase occurred in all patients who underwent distraction, and all patients showed clinical improvement. Six patients with Pierre Robin sequence became asymptomatic, with normal sleep, feeding, and weight gain. Two patients with Nager syndrome and tracheostomies were decannulated and were asymptomatic postdistraction. One patient with Treacher Collins syndrome without tracheostomy became asymptomatic after mandibular distraction; one patient failed to distract because of premature consolidation and continued to require a cannula. Mandibular distraction seems to provide a consistent change in tongue base position that improves obstructive airway symptoms by increasing measured effective airway space. The potential for mandibular distraction exceeds the simple correction of malocclusion also by eliminating soft-tissue obstruction of the micrognathic airway. Airway improvement is independent of the syndrome diagnosed. Mandibular distraction osteogenesis may be useful to avoid or decannulate existing tracheostomy in infants with micrognathia. PMID- 11496168 TI - Long-term physical impairment and functional outcomes after complex facial fractures. AB - To develop an understanding of the expected functional outcomes after facial trauma, a retrospective cohort study of patients with complex facial fractures was conducted. A cohort of adults aged 18 to 55 years who were admitted to the R. Adams Cowley Shock Trauma Center between July of 1986 and July of 1994 for treatment of a Le Fort midface fracture (resulting from blunt force) was retrospectively identified. Outcomes of interest included measures of general health status and psychosocial well being in addition to self-reported somatic symptoms. General health status was ascertained using the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36). The Body Satisfaction Scale was used to define patient concerns about altered body image and shape. To determine whether complex maxillofacial trauma and facial fractures contributed to altered social interactions, the Social Avoidance and Distress scale was used. In addition, information about a patient, his or her injury, and its treatment were ascertained from the medical records. Using the methods described above, 265 patients with Le Fort fractures were identified. These individuals were matched to a similar group of 242 general injury patients. A total of 190 of the Le Fort patients (72 percent of those eligible for the study) and 144 (60 percent) general injury patients were successfully located, and long-term interview data were acquired.Le Fort fracture patients as a group had similar health status outcomes when compared with the group of general injury patients. However, when outcomes were examined by the complexity of the Le Fort fracture, the authors found that study subjects with severe, comminuted Le Fort injuries (group D) had significantly lower SF-36 scores (worse outcomes) for the two dimensions related to role limitations: role limitations due to physical problems and role limitations due to emotional problems (p < 0.05). SF-36 scores for all other dimensions except physical function were also lower for comminuted versus less complex Le Fort fractures, although differences were not statistically significant.Specifically, there was a direct relationship between severity of facial injury and patients reporting work disability. Of group C and D Le Fort patients (severely comminuted fractures) only 55 and 58 percent, respectively, had returned to work at the time of follow-up interview. These figures are significantly lower than the back-to-work percentage of patients with less severe facial injury (70 percent). When study participants were asked if they were experiencing specific somatic symptoms at the time of the interview that they had not experienced before the injury, a significantly larger percent of the Le Fort fracture patients (compared with the general injury patients) responded in the affirmative. Differences between the Le Fort fracture and general injury groups were statistically significant (p < 0.05) for all 11 symptoms. The percentage of patients reporting complaints increased with increasing complexity of facial fracture in the areas of visual problems, alterations in smell, difficulty with mastication, difficulty with breathing, and epiphora, and these differences reached statistical significance. Patients sustaining comminuted Le Fort facial fractures report poorer health outcomes than patients with less severe facial injury and substantially worse outcomes than population norms. It is also this severely injured population that reports the greatest percentage of injury related disability, preventing employment at long-term follow-up. The long-term goal of centralized tertiary trauma treatment centers must be to return the patient to a productive, active lifestyle. PMID- 11496169 TI - Injuries associated with mandible fractures sustained in motor vehicle collisions. AB - Motor vehicle collisions are second only to altercations as the most common cause of mandible fractures. This article details in a retrospectively studied group the incidence of isolated mandible fractures and associated injuries in patients who were involved in motor vehicle collisions. This group consisted of 148 patients with mandible fractures listed in the University of Mississippi's trauma registry during the past 5 years. In almost all patients, associated injuries occurred with mandible fractures that were caused by motor vehicle collisions, with an incidence of 99.3 percent. Facial and head lacerations and facial fractures were the leading associated injuries, occurring in more than half of the patients who had a mandible fracture. Closed head injury is the major life threatening associated injury and cause of mortality. The life-threatening injuries occurred in 64.8 percent of patients in this study. The mortality rate in this group of patients was 8.1 percent. These data suggest that mandible fractures from motor vehicle collisions should never be viewed as an isolated injury but rather as part of a spectrum of significant and sometimes life threatening injuries that require thorough trauma evaluation at the time of presentation. PMID- 11496170 TI - "Axial split osteotomy" of free fibular flaps for mandible reconstruction: preliminary results. AB - Vascularized free flaps are now considered the most appropriate choice for mandible reconstruction because they offer excellent cosmetic and functional results. Various techniques have been proposed. Free fibular flaps have numerous advantages in terms of versatility and contouring. Shaping can be achieved by wedge osteotomy with excellent results. However, this technique leads to bone loss and may be difficult in the later stages of the procedure. The purpose of this report is to describe a simple and safe flap-shaping method involving axial split osteotomy. PMID- 11496171 TI - The use of multifolded pericranial flaps as "plugs" and "pads". AB - Pericranial flaps are thin and, hence, their volumes are small. Therefore, their use for soft-tissue augmentation has not been popular. In this article, the author introduces a new concept: the use of a multifolded pericranial flap as a "plug" or a "pad" for localized contour defects. Eight patients were included in the study. In all cases, an anteriorly based pericranial flap was used, and the flap was folded on itself several times to increase its bulk. The results were satisfactory in all patients. The literature on the topic is reviewed, and the blood supply of pericranial flaps is discussed. PMID- 11496173 TI - Appropriate location of the nipple-areola complex in males. AB - Gynecomastia is a common deformity encountered by plastic surgeons. The appropriate location of the nipple-areola complex is a major determinant of the aesthetic success of the procedure. To study the natural location of the nipple areola complex in the normally built male, 50 nonobese men with no evidence of gynecomastia and an average age of 27.9 years were examined. Three ratios were calculated and found to be relatively constant; they were the ratio between the height of the nipple and the height of the patient, the ratio between the distance between the nipples and chest circumference, and the ratio between the suprasternal notch-to-nipple distance and the height of the patient. Using these three parameters, a method of locating the nipple-areola complex on the male chest wall was devised. The method is advocated as a reliable, simple, and useful technique. PMID- 11496172 TI - A comparison of two methods of infiltration in breast reduction surgery. AB - The superwet technique has been shown in previous studies to dramatically reduce blood loss in breast reduction surgery, compared with standard infiltration. A retrospective chart review of 303 consecutive patients undergoing bilateral breast reduction surgery was undertaken to demonstrate additional differences in complication rate, operative time, or sponge use in the operating room. In this series, 132 consecutive patients received standard infiltration along incision lines (25 cc per breast of 1:100,000 epinephrine), and 171 patients received superwet infiltration with 240 cc per breast of 1:1,000,000 epinephrine. The average operative time was significantly reduced in the superwet group, from 78.5 minutes to 70.7 minutes (p < 0.01 level). The average number of sponges used intraoperatively was also decreased significantly (p < 0.01), from 26 to 20 sponges. Complication rates were equally low in both groups, demonstrating the safety of the superwet technique. In addition to limiting blood loss, the superwet infiltration effectively reduces operative time and sponge use without increasing complications in breast reduction surgery. PMID- 11496174 TI - Reconstructive management of contralateral breast cancer in patients who previously underwent unilateral breast reconstruction. AB - When a patient who has had unilateral breast reconstruction presents with a new cancer on the opposite side, the reconstructive management of the second breast can be unclear. This study was performed to determine whether reconstruction of the second breast is oncologically reasonable and to evaluate the reconstructive options available to these patients. Patients who had mastectomy with unilateral breast reconstruction between 1988 and 1994 and who had a minimal follow-up of 5 years from the initial breast cancer were reviewed. Of 469 patients reviewed, 18 patients (4 percent) were identified who developed contralateral breast cancer. Mean age at the initial breast cancer presentation was 43 years (range, 26 to 57 years), and mean age at presentation with contralateral breast cancer was 48 years (range, 36 to 67). The mean interval between the initial and contralateral breast cancer presentations was 5 years (range, 1 to 10 years). Mean follow-up from the time of contralateral breast cancer was 5 years (range, 1 to 9 years). In most cases, contralateral breast cancer presented at an early stage (13 of 18 patients; 72 percent), and a shift to an earlier stage at presentation of the contralateral cancer was evident compared with the initial breast cancer. Of the 18 patients who developed contralateral breast cancer, 16 (89 percent) had no evidence of disease, one was alive with disease, and one died. Reconstructive management after the initial mastectomy included 16 transverse rectus abdominis myocutaneous flaps (seven free and nine pedicled), one latissimus dorsi myocutaneous flap with implant, and one superior gluteal free flap. Surgical management of the second breast after contralateral breast cancer included breast conservation in two patients, mastectomy without reconstruction in four, and mastectomy with reconstruction in 12. Reconstruction of the second breast included one free transverse rectus abdominis myocutaneous flap, three extended latissimus dorsi flaps, two latissimus dorsi myocutaneous flaps with implants, three implants alone, two Rubens flaps, and one superior gluteal free flap. No major complications were noted after the reconstruction of the second breast. The best symmetry was obtained when similar methods and tissues were used on both sides. The incidence of contralateral breast cancer after mastectomy and unilateral breast reconstruction is low. In most cases, contralateral breast cancer presents at an earlier stage compared with the initial breast cancer, and the prognosis is good. In patients who develop a contralateral breast cancer after mastectomy and unilateral breast reconstruction, the reconstruction of the second breast after mastectomy is oncologically reasonable and should be offered to provide optimal breast symmetry and a better quality of life. The best result is obtained when similar methods and tissues are used on both sides. PMID- 11496176 TI - Nipple reconstruction using the C-V flap technique: a long-term evaluation. AB - Numerous procedures are available for nipple reconstruction with no true universal favorite. This study presents long-term follow-up data for nipple reconstruction using the C-V flap technique. Patients were identified by searching the Breast Reconstruction Database, and they were asked to return for a follow-up visit. All those who underwent nipple reconstruction using the C-V flap technique between January of 1992 and December of 1996 were reviewed in an attempt to conduct a long-term follow-up evaluation. The response was poor, and 11 patients participated in the study and returned for follow-up. They all completed a questionnaire, which focused on patient satisfaction using a visual analogue scale. Nipple measurements were taken with a caliper and compared with the opposite breast for symmetry. Fourteen nipple reconstructions were evaluated in 11 patients with an average follow-up of 5.3 years. All patients had undergone transverse rectus abdominis musculocutaneous (TRAM) flap reconstructions. Patient satisfaction was 42 percent with nipple projection, 62 percent with pigmentation, and 26 percent with sensation. Overall patient satisfaction with the procedure was 81 percent. Average nipple projection of the reconstructed nipple was 3.77 mm and was not statistically different when compared with the opposite nipple. Long term subjective evaluation of the C-V flap technique does report a loss in nipple projection; however, overall patient satisfaction at 5.3 years is good, as is the ability to restore symmetry with the opposite breast. PMID- 11496177 TI - An Internet-based utility assessment of breast hypertrophy. AB - The Internet has changed the way people shop, do business, and communicate with one another. Even those who try to avoid computers are bombarded with Web advertisements on television and in newspapers and magazines. The Internet also has excellent, but still largely underused, potential for conducting research studies. A Web-based questionnaire essentially combines the power and graphic flexibility of a computer with the freedom of a mail survey. The specific goals of this study were (1) to develop a Web-based utility assessment tool and (2) to use this tool to measure the health burden of breast hypertrophy by using the Internet to sample a population. An open-enrollment, Internet-based survey was developed to assess societal preferences for mild and severe breast hypertrophy using three established assessment techniques: visual analogue scale, time trade off, and standard gamble. Subjects were recruited from a Web-based clinical trial listing service. Demographics, subjects' utility for their current health, and responses to a comorbidity index were also recorded. Data were recorded from August 1, 1999, to January 31, 2000. There were 480 unique responses, and 356 (74 percent) met the inclusion criteria. The respondents were predominantly female (81 percent), Caucasian (83 percent), and in the middle income brackets. Their mean age was 32.9. The average score for capacity of understanding was 4.99 out of 5 (5 = excellent). The median utility score for severe breast hypertrophy (visual analogue scale, 0.70; time trade-off, 0.85; standard gamble, 0.88) differed significantly from the median utility score for mild breast hypertrophy (visual analogue scale, 0.93; time trade-off, 1.0; standard gamble, 0.98) for each method. The results showed that the construction of a Web-based questionnaire for utility assessment is feasible and can be used to capture the utility of health states. The authors were able to enroll a large number of subjects with excellent capacity to understand the study, resulting in a high rate of usable responses. The applicability of these data to cost-effectiveness studies is limited by the extent to which the sampled population of this study is representative of society in general. The demographics of this study sample also differed from those of the Internet population. The study was piloted by measuring the values for breast hypertrophy, but the procedure could be used to assess the burden on quality of life of any disease and, potentially, the efficacy of surgical interventions. The study method is recommended as an accurate and cost-effective alternative for measuring quality of life. PMID- 11496178 TI - Reconstruction of burn scar of the upper extremities with artificial skin. AB - The management of upper-extremity burn contractures is a major challenge for plastic surgeons. After approval by the Food and Drug Administration, artificial skin (Integra) has been available in Taiwan since 1997. From January of 1997 to July of 1999, the authors applied artificial skin to 13 severely burned patients for the reconstruction of their upper extremities, resulting in an increased range of motion in the upper-extremity joints and improved skin quality. An additional benefit was the rapid reepithelialization of the donor sites. There were no complications of infection throughout the therapeutic course, and the overall results were satisfactory. During the 2-year study, scar condition was monitored between 8 and 24 months, and a good appearance and pliable skin were obtained according to the Vancouver Scar Scale. According to this evaluation of Oriental skin turgor, normal pigmentation was restored about 6 months after the resurfacing procedure. For patients with severe burns in whom there is insufficient available skin for a full-thickness skin graft or another appropriate flap for scar revision, Integra is an alternative. The two major concerns in dealing with artificial skin are (1) a 10- to 14-day waiting period for maturation of the neo-dermis, necessitating a two-stage operation, and (2) prevention of infection with antibiotics and meticulous wound care. PMID- 11496180 TI - Early passive mobilization after digital nerve repair and grafting in a fresh cadaver. AB - In the clinical management of combined tendon and nerve injuries, there are competing treatment strategies. Isolated tendon injuries should be rapidly mobilized after repair to prevent adhesion formation, whereas isolated nerve repairs are usually immobilized to prevent disruption and to allow axon regrowth. Recommendations in the published literature for the management of combined tendon and nerve injuries are vague and advise up to 3 weeks of immobilization. The goals of this study were to determine which length of nerve gap resulted in rupture of a repair following postoperative mobilization with the modified Duran protocol and with unrestricted motion and to determine whether nerve grafts are at risk of rupture after mobilization. A total of 100 digital nerves from 10 cadaver hands were tested with the modified Duran and the unsplinted protocols. Each digital nerve on each hand was sequentially resected and repaired at five progressively larger gap lengths after testing with both protocols. The mean nerve gaps at which disruption occurred were significantly different between the splinted (9.7 +/- 0.8 mm, n = 100) and unsplinted (7.3 +/- 1.9 mm, n = 100) protocols (t test, p < 0.001). One hundred percent of repairs remained intact, with up to 5 mm of resection with the modified Duran protocol (n = 100) and with up to 2.5 mm of resection with the unsplinted protocol (n = 100). All nerve grafts remained intact after mobilization within a dorsal-blocking splint (n = 100). Considering mechanical integrity of the nerve repair only, these data suggest that early mobilization with tendon protocols may be considered after a nerve injury to avoid the detrimental tendon sequelae that result from immobilization. The adequacy of functional recovery of mobilized nerves is yet to be determined. PMID- 11496181 TI - Effects of cell concentration and growth period on articular and ear chondrocyte transplants for tissue engineering. AB - This study determined the effects of chondrocyte source, cell concentration, and growth period on cartilage production when isolated porcine cells are injected subcutaneously in a nude mouse model. Chondrocytes were isolated from both ear and articular cartilage and were suspended in Ham's F-12 medium at concentrations of 10, 20, 40, and 80 million cells per cubic centimeter. Using the nude mouse model, each concentration group was injected subcutaneously in 100-microl aliquots and was allowed to incubate for 6 weeks in vivo. In addition, cells suspended at a fixed concentration of 40 million cells per cubic centimeter were injected in 100-microl aliquots and were incubated for 1, 2, 3, 4, 5, 6, 9, and 12 weeks. Each concentration or time period studied contained a total of eight mice, with four samples harvested per mouse for a final sample size of 32 constructs. All neocartilage samples were analyzed by histologic characteristics, mass, glycosaminoglycan level, and DNA content. Control groups consisted of native porcine ear and articular cartilage. Specimen mass increased with increasing concentration and incubation time. Ear neocartilage was larger than articular neocartilage at each concentration and time period. At 40 million cells per cubic centimeter, both ear and articular chondrocytes produced optimal neocartilage, without limitations in growth. Specimen mass increased with incubation time up to 6 weeks in both ear and articular samples. No significant variations in glycosaminoglycan content were found in either articular or ear neocartilage, with respect to variable chondrocyte concentration or growth period. Although articular samples demonstrated no significant trends in DNA content over time, ear specimens showed decreasing values through 6 weeks, inversely proportional to increase in specimen mass. Although both articular and ear sources of chondrocytes have been used in past tissue-engineering studies with success, this study indicates that a suspension of ear chondrocytes injected into a subcutaneous location will produce biochemical and histologic data with greater similarity to those of native cartilage. The authors believe that this phenomenon is attributable to the local environment in which isolated chondrocytes from different sources are introduced. The subcutaneous environment of native ear cartilage accommodates subcutaneously injected ear chondrocyte transplants better than articular transplants. Native structural and biochemical cues within the local environment are believed to guide the proliferation of the differentiated chondrocytes. PMID- 11496182 TI - An in vitro mouse model of cleft palate: defining a critical intershelf distance necessary for palatal clefting. AB - It is unclear whether cleft palate formation is attributable to intrinsic biomolecular defects in the embryonic elevating palatal shelves or to an inability of the shelves to overcome a mechanical obstruction (such as the tongue in Pierre Robin sequence) to normal fusion. Regardless of the specific mechanism, presumably embryonic palatal shelves are ultimately unable to bridge a critical distance and remain unapproximated, resulting in a clefting defect at birth. We propose to use a palate organ culture system to determine the critical distance beyond which embryonic palatal shelves fail to fuse (i.e., the minimal critical intershelf distance). In doing so, we hope to establish an in vitro cleft palate model that could then be used to investigate the contributions of various signaling pathways to cleft formation and to study novel in utero treatment strategies. Palatal shelves from CD-1 mouse embryos were microdissected on day 13.5 of gestation (E13.5; term = 19.5 days), before fusion. Using a standardized microscope ocular grid, paired palatal shelves were placed on a filter insert at precisely graded distances ranging from 0 (in contact) to 1.9 mm (0, 0.095, 0.19, 0.26, 0.38, 0.48, 0.57, 0.76, 0.95, and 1.9 mm). A total of 68 paired palatal shelves were placed in serum-free organ culture for 96 hours (n = 68). Sample sizes of 10 were used for each intershelf distance up to and including 0.48 mm (n = 60). For intershelf distances of 0.57 mm and greater, two-paired palatal shelves were cultured (n = 8). All specimens were assessed grossly and histologically for palatal fusion. Palatal fusion occurred in our model only when intershelf distances were 0.38 mm or less. At 0.38 mm, eight of 10 palates appeared grossly adherent, whereas six of 10 demonstrated clear fusion histologically with resolution of the medial epithelial seam and continuity of the palatal mesenchyme. None of the 18 palates fused when placed at intershelf distances of 0.48 mm or greater. Using our selected intershelf distances as a guideline, we have established an approximate minimal critical intershelf distance (0.48 mm) at which we can reliably expect no palatal fusion. Culturing palatal shelves at intershelf distances of 0.48 mm or greater results in nonfusion or clefting in vitro. This model will allow us to study biomolecular characteristics of unfused or cleft palatal shelves in comparison with fused shelves. Furthermore, we plan to study the efficacy of grafting with exogenous embryonic mesenchyme or candidate factors to overcome clefting in vitro as a first step toward future in utero treatment strategies. PMID- 11496183 TI - Influence of different biodegradable carriers on the in vivo behavior of human adipose precursor cells. AB - The correction of soft-tissue defects presents a challenge in plastic and reconstructive surgery. The implantation of isolated and culture-expanded adipose precursor cells is a solution to this problem because these cells differentiate into adipocytes when implanted in vivo. Appropriate scaffolds are needed in soft tissue engineering to allow the differentiation of precursor cells. The optimal carrier needs to be defined. In this study, human preadipocytes were isolated and cultured. Three different carrier materials were seeded with 106 preadipocytes each and implanted in 42 nude mice. Sponges and nonwoven carriers based on hyaluronic acid modified by esterification (HYAFF 11) were compared with collagen sponges. Scaffolds without cells served as negative controls in the same animal. After 3 and 8 weeks, the grafts were explanted. Macroscopic appearance, weight, thickness, microscopy, immunohistochemistry, and TEM (scaffold structure, cellularity, penetration depth of the seeded cells, vascularization) were assessed and evaluated for differences in scaffold-cell interactions.Preadipocytes differentiated earlier in vitro when attached to HYAFF 11 scaffolds than to other carrier materials. Macroscopically, all preadipocyte constructs were yellowish and well vascularized, and the controls were white and avascular. Vessel formation was more pronounced around mature adipocytes. Microscopically, HYAFF 11 constructs showed a higher cell density than collagen constructs. The pores of the sponges contained more differentiated adipocytes than the nonwoven carriers, whereas the undifferentiated preadipocytes were more numerous in the nonwoven material. Penetration of adipose precursor cells was deeper and more homogeneous in HYAFF 11 scaffolds. Electron microscopy demonstrated well-differentiated adipocytes and large amounts of extracellular matrix in HYAFF 11 sponges.HYAFF 11 sponges supported the expansion and differentiation of the adipose precursor cells. This carrier is superior to the nonwoven carrier with regard to adipocyte differentiation and superior to the collagen sponge with regard to cellularity. This is a promising method for the reconstruction of soft-tissue defects. Modifications of the scaffold (larger pore size and coating with adipogenic factors) will be examined in further experiments. PMID- 11496185 TI - Differential expression of transforming growth factor-beta receptors I and II and activation of Smad 3 in keloid fibroblasts. AB - Keloids represent a dysregulated response to cutaneous wounding that results in an excessive deposition of extracellular matrix, especially collagen. However, the molecular mechanisms regulating this pathologic collagen deposition still remain to be elucidated. A previous study by this group demonstrated that transforming growth factor (TGF)-beta1 and -beta2 ligands were expressed at greater levels in keloid fibroblasts when compared with normal human dermal fibroblasts (NHDFs), suggesting that TGF-beta may play a fibrosis-promoting role in keloid pathogenesis.To explore the biomolecular mechanisms of TGF-beta in keloid formation, the authors first compared the expression levels of the type I and type II TGF-beta receptors in keloid fibroblasts and NHDFs. Next, they investigated the phosphorylation of Smad 3, an intracellular TGF-beta signaling molecule, in keloid fibroblasts and NHDFs. Finally, they examined the regulation of TGF-beta receptor II by TGF-beta1, TGF-beta2, and TGF-beta3 ligands. Our findings demonstrated an increased expression of TGF-beta receptors (types I and II) and increased phosphorylation of Smad 3 in keloid fibroblasts relative to NHDFs. These data support a possible role of TGF-beta and its receptors as fibrosis-inducing growth factors in keloids. In addition, all three isoforms of recombinant human TGF-beta proteins could further stimulate the expression of TGF beta receptor II in both keloids and NHDFs. Taken together, these results substantiate the hypothesis that the elevated levels of TGF-beta ligands and receptors present in keloids may support increased signaling and a potential role for TGF-beta in keloid pathogenesis. PMID- 11496186 TI - Effect of total venous occlusion on capillary flow and necrosis in skeletal muscle. AB - Limb replantation and microvascular transfer of flaps are sometimes complicated by postoperative venous thrombosis. Total venous occlusion can lead to complete shutdown of microvascular perfusion, resulting in failure of the transfer or replantation. Once venous return stops, it must be restored within a critical period of time for tissue survival. The purpose of this experiment was to delineate this critical period of time at which no reflow and irreversible muscle necrosis occurs by the use of a rat gracilis flap microcirculation model. The gracilis muscle of 40 male Wistar rats (135.3 +/- 37.2 g) was elevated on its vascular pedicle and mounted on a raised platform for videomicroscopic analysis. Animals were randomly assigned to one of four groups: (1) sham (no total venous occlusion), (2) 10 minutes of total venous occlusion, (3) 30 minutes of total venous occlusion, and (4) 60 minutes of total venous occlusion. Total venous occlusion was established by placing a microvascular clamp across the femoral vein at the junction of the gracilis pedicle. The number of flowing capillaries in five consecutive high-power fields (832x) were counted at baseline and at 5, 15, 30, 60, 120, 180 minutes, and 24 hours after reperfusion. At 24 hours after reperfusion, the gracilis muscles were harvested and stained with nitroblue tetrazolium. Percentage of muscle necrosis was measured by using computer planimetry. The data were reported as mean +/- standard error of mean and were compared between groups by analysis of variance and appropriate post hoc comparisons. Total venous occlusion for 10, 30, and 60 minutes showed a significant decrease in the number of flowing capillaries through 24-hour postreversal. There was a significant drop (p < 0.01) in the number of flowing capillaries from 30 minutes of total venous occlusion to 60 minutes of total venous occlusion at all times. Muscle necrosis was significantly increased in all three groups of total venous occlusion compared with the sham group (36.1 +/- 1.7 percent, 45.5 +/- 3.4 percent, 74.1 +/- 4.7 percent versus 14.3 +/- 1.7 percent, and p < 0.01). These results indicate that irreversible tissue damage occurs in a very short time interval (60 minutes) in this model, making the early detection of venous occlusion critical to the successful correction of this complication. PMID- 11496187 TI - Ectopic bone formation in titanium mesh loaded with bone morphogenetic protein and coated with calcium phosphate. AB - The osteoinductive properties of porous titanium fiber mesh, with or without a calcium phosphate coating and loaded with recombinant human bone morphogenic protein-2 (rhBMP-2) or rhBMP-2 and native bovine BMP (S-300) were investigated in a rat ectopic assay model. A total of 112 calcium phosphate-coated and 112 noncoated porous titanium implants, either loaded with rhBMP-2 and S-300 or loaded with rhBMP-2 alone, were subcutaneously placed in 56 Wistar-King rats. The rats were killed 5, 10, 20, and 40 days postoperatively, and the implants were retrieved. Histologic analysis demonstrated that all growth factor and carrier combinations induced ectopic cartilage and bone formation at 5 and 10 days, respectively. At 20 days, bone formation increased and was characterized by trabecular bone and bone marrow-like tissue. At 40 days, more lamellar bone and hemopoietic bone marrow-like tissue were present. At both times, more bone had been formed in calcium phosphate-coated implants than in noncoated samples. Further, in rhBMP-2 and S-300-loaded specimens, bone formation was higher than in rhBMP-2 only-loaded specimens. In rhBMP-2 only-loaded specimens, bone formation was mainly localized inside the mesh material, whereas in specimens loaded with both rhBMP-2 and S-300, the bone was localized inside and surrounding the titanium mesh. The histological findings were confirmed by calcium content and alkaline phosphatase activity measurements. In addition, all specimens showed osteocalcin expression as early as 5 days postoperatively. Our results show that the combination of titanium mesh with BMPs can induce ectopic bone formation and that this bone formation seems to be similar to "enchondral" ossification. In addition, a thin calcium phosphate coating can have a beneficial effect on the bone-inducing properties of a scaffold material. Finally, rhBMP-2 and native BMP act synergistically in ectopic bone induction. PMID- 11496188 TI - Free tissue transfer in a ventricular assist device patient. PMID- 11496189 TI - Thin paraumbilical perforator-based cutaneous island flap for scrotal resurfacing. PMID- 11496190 TI - A newly designed nasal retainer to sustain the corrected shape of the nostril rim and anterior nasal recess for cleft lip patients. PMID- 11496191 TI - A novel method for repair of flexor digitorum superficialis lacerations in zone 2. PMID- 11496192 TI - Web-based medicine as a means to establish centers of surgical excellence in the developing world. AB - The growth of the Internet has provided a unique opportunity for rapid, global communication. Web-based medicine uses this technology to help surgeons in developing regions of the world gain direct access to recognized experts. This serves to empower local surgeons in the developing world through direct skill transfer and encouraging academic pursuit. Web-based medicine follows the paradigm of a university without borders, therefore requiring exacting patient record keeping, monthly peer review, and continuing medical education of all its participants. All those who participate in Web-based medicine have undergone a credentialing process to guarantee that they possess adequate credentials. Patient confidentiality is strictly maintained. Web-based medicine also provides a follow-up strategy for medical volunteer groups who provide overseas services. Interplast, Inc., has administered a Web-based medicine site at http://www.wiredmd.com since July of 1999. A total of 767 cleft malformation cases performed locally by participating host surgeons in the developing world have been reviewed through the site. Additionally, 16 consultations have been posted and discussed by participating surgeons worldwide. Financing remains the major impediment to the globalization of this technology. PMID- 11496193 TI - The power of PowerPoint. AB - Carousel slide presentations have been used for academic and clinical presentations since the late 1950s. However, advances in computer technology have caused a paradigm shift, and digital presentations are quickly becoming standard for clinical presentations. The advantages of digital presentations include cost savings; portability; easy updating capability; Internet access; multimedia functions, such as animation, pictures, video, and sound; and customization to augment audience interest and attention. Microsoft PowerPoint has emerged as the most popular digital presentation software and is currently used by many practitioners with and without significant computer expertise. The user-friendly platform of PowerPoint enables even the novice presenter to incorporate digital presentations into his or her profession. PowerPoint offers many advanced options that, with a minimal investment of time, can be used to create more interactive and professional presentations for lectures, patient education, and marketing. Examples of advanced PowerPoint applications are presented in a stepwise manner to unveil the full power of PowerPoint. By incorporating these techniques, medical practitioners can easily personalize, customize, and enhance their PowerPoint presentations. Complications, pitfalls, and caveats are discussed to detour and prevent misadventures in digital presentations. Relevant Web sites are listed to further update, customize, and communicate PowerPoint techniques. PMID- 11496194 TI - The L short-scar mammaplasty: 12 years later. PMID- 11496195 TI - Reconstruction of the cheek. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Describe the cheek unit and its specific requirements for quality, outline, and contour. 2. Identify factors involving the patient, wound, and donor materials that determine technique. 3. Understand the application of anteriorly based, posteriorly based, and advancement flaps, and the use of serial excisions for specific defects. 4. Identify, prevent, and treat complications of cheek reconstruction. PMID- 11496196 TI - Mirror, mirror on the wall: when the postoperative reflection does not meet patients' expectations. PMID- 11496197 TI - Improvements in cardiovascular risk profile with large-volume liposuction: a pilot study. AB - In this study, the authors investigated the physiologic effects of the altered body composition that results from surgical removal of large amounts of subcutaneous adipose tissue. Fourteen women with body mass indexes of greater than > 27 kg/m2 underwent measurements of fasting plasma insulin, triglycerides, cholesterol, body composition by dual-energy x-ray absorptiometry (DXA), resting energy expenditure, and blood pressure before and after undergoing large-volume ultrasound-assisted liposuction. There were no significant intraoperative complications. Body weight had decreased by 5.1 kg (p < 0.0001) by 6 weeks after liposuction, with an additional 1.3-kg weight loss (p < 0.05) observed between 6 weeks and 4 months after surgery, for a total weight loss of 6.5 kg (p < 0.00006). Body mass index decreased from (mean +/- SEM) 28.8 +/- 2.3 to 26.8 +/- 1.5 kg/m2 (p < 0.0001). This change in body weight was primarily the result of decreases in body fat mass: as assessed by DXA, lean body mass did not change (43.8 +/- 3.1 kg to 43.4 +/- 3.6 kg, p = 0.80), whereas DXA total body fat mass decreased from 35.7 +/- 6.3 to 30.1 +/- 6.5 kg (p < 0.0001). There were significant decreases in fasting plasma insulin levels (14.9 +/- 6.5 mIU/ml before liposuction versus 7.2 +/- 3.2 mIU/ml 4 months after liposuction, p < 0.007), and systolic blood pressure (132.1 +/- 7.2 versus 120.5 +/- 7.8 mmHg, p < 0.0002). Total cholesterol, high-density lipoprotein cholesterol, plasma triglycerides, and resting energy expenditure values were not significantly altered after liposuction. In conclusion, over a 4-month period, large-volume liposuction decreased weight, body fat mass, systolic blood pressure, and fasting insulin levels without detrimental effects on lean body mass, bone mass, resting energy expenditure, or lipid profiles. Should these improvements be maintained over time, liposuction may prove to be a valuable tool for reducing the comorbid conditions associated with obesity. PMID- 11496199 TI - Importance of the nasal-to-cervical relationship to the profile in rhinoplasty surgery. AB - There is general agreement that when discussing surgery with the prospective rhinoplasty patient, one may also include discussion of the chin because of the important interrelationship between these two regions. It is apparent that on the profile-lateral view, the four prominences-the forehead, nose, chin, and neck balance and complement one another. The cervical region, the fourth dimension, was examined to estimate the aesthetic significance of the nasal-to-cervical relationship and to determine the implications to rhinoplasty surgery. Part I of the study was a survey to test the hypothesis that the cervical region affects the perceived impression of the nose. Four standard facial-profile black-and white photographs were chosen to represent varying degrees of nasal dorsum hump and cervical ptosis problems. Using computer-altering software, only the cervical regions were altered to create a pair of photographs for each of the four profiles: one with a youthful-looking non-ptotic cervical region, the other with a ptotic aged-looking cervical region. Raters were asked to give their first impression opinions of which nose subjectively appeared "better." Raters consistently (84 percent of the time) chose the nose on the faces with the less ptotic neck as being better. Therefore, a less ptotic neck improved the perceived appearance of the nose. Part II was a retrospective chart review of the rhinoplasty patients of a single surgeon by independent raters. To estimate the aesthetic significance of the nasal-to-cervical relationship, the proportion of patients undergoing rhinoplasty surgery who could have potentially benefited from a youth-restoring neck procedure was determined. Criteria originally described by Ellenbogen and Karlin for judging the results of youth-restoring neck procedures were used as relative indications for neck surgery. An average of 27.2 percent of the patients did not have visible criteria and therefore by definition had relative indications for neck-rejuvenating procedures when undergoing rhinoplasty surgery. As demonstrated in part I of the study, improving the neck could improve the perceived results of the rhinoplasty. Part III of the study validated the Ellenbogen and Karlin criteria. The present authors found that the original criteria were probably based on female patients, that male and older patients had more indications for surgery, and that there was significant interrater agreement with the youthful criteria. In summary, the authors established that a strong nasal-to-cervical relationship exists whereby the perceived appearance of the nose is affected by the neck. The significance of this relationship to rhinoplasty surgery was determined, and it was found that more than 27 percent of rhinoplasty patients could obtain better perceived nasal results with a concomitant neck-rejuvenating procedure. Consequently, discussing neck rejuvenating procedures with the rhinoplasty patient is valuable. PMID- 11496202 TI - Rationale for submucous resection of hypertrophied inferior turbinates in rhinoplasty: an evolution. AB - To achieve success in rhinoplasty, the plastic surgeon takes advantage of numerous intraoperative techniques designed to manipulate nasal soft tissue and the osseocartilaginous framework. Although the postoperative result may meet preoperative aesthetic goals, an element of nasal airway obstruction can persist from failure to acknowledge the role of inferior turbinates. Surgically responsive inferior turbinate hypertrophy is frequently not addressed secondary to inadequate history taking, incomplete physical examination, and/or surgeon reluctance to handle these sensitive structures. The goal of this article is to discuss the anatomy and physiology of the inferior turbinates, to present the role for inferior turbinate surgery during rhinoplasty, and to delineate the evolution of the current technique of submucosal resection of the inferior turbinates. Over the past 14 years, the senior author (R.J.R.) has performed inferior turbinates surgery on 648 patients as part of a rhinoplasty. PMID- 11496204 TI - Lengthening the foreshortened nose. AB - The surgically shortened nose is an infrequent but distressing problem that is difficult to repair. This article describes a simple technique to lengthen the nasal profile with a triple-stacked tip graft placed caudal to the medial crura. Fifteen patients have been operated on, and most were undergoing face lifts. The skin inelasticity in older patients allows easy skin redraping over the larger tip volume. PMID- 11496205 TI - Advancement of the external oblique muscle flap to improve the waistline: a study in cadavers. AB - Waist definition is an important goal of abdominoplasty. The purpose of this report is to study the effect of advancement and rotation of the external oblique muscles in the waistline. Twenty cadavers were dissected. Two parameters were analyzed: measurement of the waist circumference and the width of overlapped skin flaps of the abdomen in the midline. Three stages of dissection were compared: (1) initial stage, in which the myoaponeurotic structure of the anterior abdominal wall was intact; (2) stage 1, after advancement of the rectus muscles and its anterior aponeurosis toward the midline; and (3) stage 2, after advancement and rotation of the external oblique muscle. A significant statistical difference was found when waist circumference measured before dissection was compared with values obtained after the procedure. When the width of the overlapped skin flaps was compared during the different stages of dissection, a significant statistical difference was observed after each stage (paired t test). Therefore, the approximation of the rectus muscles alone improves the waistline, and when associated with external oblique muscle flap advancement and rotation, the cosmetic result in this area is even better. In conclusion, the procedure described decreases waist circumference and improves the frontal view of the waist. PMID- 11496208 TI - Limited incision forehead lift for eyebrow elevation to enhance upper blepharoplasty. PMID- 11496206 TI - Body mass index: risk predictor for cosmetic day surgery. AB - Body mass index (BMI; weight per unit surface area) is the scientific yardstick by which overweight is gauged relative to the population norm. The contrary association between obesity and diabetes or hypertension is only too well known. Less appreciated is the heightened sensitivity to respiratory depressants such as sedatives and analgesics in the obese (BMI >/= 30) and the increased incidence of sleep apnea in the morbidly obese (BMI >/= 35)-either or both of which raise the risk of cosmetic surgery when sedation or anesthesia is contemplated. Guided by the BMI, a gender-independent measure of fatness, the surgeon now can inform the patient of her or his relative operative risk and offer an objective rationale for advising overnight hospitalization rather than office-based day surgery. The BMI is readily calculated when height and weight are expressed in metric units, much less so when measured in foot-pound units. In fact, the calculations are sufficiently cumbersome that the BMI remains underused in U.S. office surgery. The author's complimentary "BMI Calculator"-an Excel workbook available on-line to society members-is designed so that office staff need enter only height (in feet and inches) and weight (in pounds) to print the BMI for the patient's permanent record. The BMI places patient weight relative to height in proper perspective for aesthetic surgery, whether with sedation or under general anesthesia. The BMI ought to be as routine a part of the preoperative assessment as blood pressure or hemoglobin content. PMID- 11496209 TI - Plastic surgeon: asset or mere mercenary? PMID- 11496210 TI - The unpardonable "success fee". PMID- 11496211 TI - Will Fas gene help to diagnose burn scar squamous cell carcinoma? PMID- 11496212 TI - Postburn contractures. PMID- 11496213 TI - Rotated osteotome. PMID- 11496214 TI - Breast reconstruction with implants and expanders. PMID- 11496215 TI - Combined liposuction with abdominoplasty. PMID- 11496216 TI - Augmentation mammaplasty and use of a "tilt test". PMID- 11496217 TI - Salvage by heparinized blood injection through the alternate pedicle in a toe transplant. PMID- 11496219 TI - Further musings of an older timer. PMID- 11496218 TI - Plastic surgery from Cambridge, U.K., to Cambridge, Massachusetts, U.S.A. PMID- 11496220 TI - Molecular assessment of head and neck cancer: the difficult passage from basic science to clinical application. PMID- 11496221 TI - Internal mammary artery pedicled island flap for the treatment of chest wall radionecrosis. PMID- 11496222 TI - Bifid median nerve. PMID- 11496223 TI - Healing of diabetic ulcers with granulated sugar. PMID- 11496224 TI - The expert witness. PMID- 11496225 TI - Why suture pectoral edges in submuscular augmentation mammaplasty? PMID- 11496226 TI - Positioning of the nasal tip. PMID- 11496227 TI - Flap stretching for management of postburn knee contracture with unstable scar. PMID- 11496229 TI - Endotoxin-stimulated innate immunity: A contributing factor for asthma. AB - Exposure to airborne endotoxin in infancy may protect against asthma by promoting enhanced T(H)1 response and tolerance to allergens. On the other hand, later in life, it adversely affects patients with asthma. Endotoxin binding to receptors on macrophages and other cells generates IL-12, which inhibits IgE responses. It also generates cytokines like IL-1, TNF-alpha, and IL-8, which cause inflammation. These signal transduction pathways resemble those leading to the generation of cytokines, such as IL-4, IL-13, and IL-5, which are responsible for the inflammation of IgE-mediated allergic disease. The main difference seems to be that endotoxin recruits neutrophils, but IgE recruits eosinophils, and the details of the tissue injury from these granulocytes differ. Sources of airborne endotoxin include many agricultural dusts, aerosols from contaminated water in many industrial plants, contaminated heating and air-conditioning systems, mist generating humidifiers, and damp or water-damaged homes. Acute inhalation of high concentrations of endotoxin can cause fever, cough, and dyspnea. Chronic inhalation of lesser amounts causes chronic bronchitis and emphysema and is associated with airway hyperresponsiveness. Airborne endotoxin adversely affects patients with asthma in 3 ways: (1) by increasing the severity of the airway inflammation; (2) by increasing the susceptibility to rhinovirus-induced colds; and (3) by causing chronic bronchitis and emphysema with development of irreversible airway obstruction after chronic exposure of adults. The most effective management is mitigating exposure. The potential of drug treatments requires further clinical investigation. PMID- 11496230 TI - Genetics of allergen-induced asthma. AB - Antigen-induced airway hyperresponsiveness and airway inflammation are features of both human asthma and animal models of this disease. The genesis of these key asthma phenotypes represents the summation of a complex cascade of immune responses. It is hypothesized that multiple cell types are involved in the induction, propagation, and maintenance of these immune processes. Several molecules have been reported to be essential for cell-cell interactions, inflammatory cell recruitment, and effector functions leading to the overall expression of the asthmatic phenotype. This review summarizes the genetic evidence supporting a role for these molecules in antigen-driven airway hyperresponsiveness and inflammation. PMID- 11496231 TI - The relationship of asthma therapy and Churg-Strauss syndrome: NIH workshop summary report. AB - The Churg-Strauss syndrome (CSS) is a distinct form of vasculitis that is notable for its eosinophilia and frequent associations with asthma and sinusitis. Because there has been an increasing recognition that CSS can develop in patients with asthma and that CSS might be associated with specific asthma treatments, the National Heart, Lung, and Blood Institute, the National Institute of Allergy and Infectious Diseases, the Office of Rare Diseases, National Institutes of Health, and the US Food and Drug Administration jointly sponsored a workshop to consider interrelationships among CSS, asthma, and asthma therapeutics and to assess what is known about underlying mechanisms of CSS. Issues related to the criteria for defining and diagnosing CSS were reviewed, including the contemporary understanding that diagnostic biopsies need only reveal eosinophilic perivascular infiltrates and that asthma need not be present when CSS develops. From published reports and reports to the US Food and Drug Administration, treatment of patients with asthma with any of 3 cysteinyl leukotriene receptor antagonists, a 5 lipoxygenase inhibitor, and inhaled corticosteroids has been associated with CSS development. It is unknown whether these agents were eliciting CSS. A variety of physiologic and study design issues might lead to the reported associations of these drugs with CSS. Because many asthma patients receiving these therapies were able to diminish their systemic corticosteroid therapy, it is possible that incipient CSS was unmasked by lessened steroid use. The underlying pathophysiologic mechanisms of CSS, however, are unknown, and there is no means of identifying which patients with asthma might be at risk for CSS. Accordingly, investigations with the goals of defining the underlying pathophysiologic processes of CSS and establishing the relationships of asthma and its therapies to CSS are needed. PMID- 11496232 TI - Omalizumab, anti-IgE recombinant humanized monoclonal antibody, for the treatment of severe allergic asthma. AB - BACKGROUND: A recombinant humanized anti-IgE mAb, omalizumab, forms complexes with free IgE, blocking its interaction with mast cells and basophils; as a consequence, it might be effective in the treatment of asthma. OBJECTIVE: The purpose of this study was to evaluate the efficacy and safety of omalizumab in the treatment of inhaled corticosteroid-dependent asthma. METHODS: In this phase III, double-blinded, placebo-controlled trial, 525 subjects with severe allergic asthma requiring daily inhaled corticosteroids were randomized to receive placebo or omalizumab subcutaneously every 2 or 4 weeks, depending on baseline IgE level and body weight. Inhaled corticosteroid doses were kept stable over the initial 16 weeks of treatment and tapered during a further 12-week treatment period. RESULTS: Omalizumab treatment resulted in significantly fewer asthma exacerbations per subject and in lower percentages of subjects experiencing an exacerbation than placebo treatment during the stable steroid phase (0.28 vs 0.54 [P =.006] and 14.6% vs 23.3% [P =.009], respectively) and during the steroid reduction phase (0.39 vs 0.66 [P =.003] and 21.3% vs 32.3% [P =.004], respectively). Beclomethasone dipropionate reduction was significantly greater with omalizumab treatment than with placebo (median 75% vs 50% [P <.001]), and beclomethasone dipropionate discontinuation was more likely with omalizumab (39.6% vs 19.1% [P <.001]). Improvements in asthma symptoms and pulmonary function occurred along with a reduction in rescue beta-agonist use. Omalizumab was well tolerated, with an adverse-events profile similar to that of placebo. CONCLUSION: The addition of omalizumab to standard asthma therapy reduces asthma exacerbations and decreases inhaled corticosteroid and rescue medication use. PMID- 11496233 TI - Immunostimulatory sequence DNA linked to the Amb a 1 allergen promotes T(H)1 cytokine expression while downregulating T(H)2 cytokine expression in PBMCs from human patients with ragweed allergy. AB - BACKGROUND: Recent studies have demonstrated that bacterially derived immunostimulatory sequences (ISSs) of DNA can activate the mammalian innate immune system and promote the development of T(H)1 cells. Promotion of T(H)1 immunity by means of immunotherapy in allergic patients has led to the alleviation of symptoms that result from allergen-specific T(H)2 responses. OBJECTIVE: Our purpose was to investigate whether the T(H)1-enhancing properties of ISSs could be used to alter the T(H)2-dominated immune response of allergic PBMCs in vitro. METHODS: Ragweed protein-linked ISS (PLI) was generated from a specific, highly active 22-base ISS and Amb a 1, the immunodominant allergen in ragweed pollen, to combine the T(H)1-enhancing properties of ISSs with allergen selectivity, and its activity was investigated in PBMC cultures from subjects with ragweed allergy. RESULTS: PLI was markedly successful at reversing the dominant allergen-induced T(H)2 profile while greatly enhancing IFN-gamma production. Delivering ISSs in a linked form proved to be much more effective at modulating the resulting cytokine profile than delivering free ISSs in a mixture with unlinked Amb a 1. PLI also demonstrated cytokine-modulating properties, even when used to stimulate cells that had already been primed for 6 days with Amb a 1. The antigen specificity of the action of PLI was confirmed by the observations that PLI enhances Amb a 1--specific T-cell proliferation. CONCLUSION: These data indicate that delivery of ISSs within an antigen-specific context exhibits potent cytokine-modulating activity and, combined with its reduced allergenicity, makes this molecule a strong candidate for use in improved immunotherapy applications. PMID- 11496234 TI - Allergen-induced impairment of bronchoprotective nitric oxide synthesis in asthma. AB - BACKGROUND: Endogenous nitric oxide protects against airway hyperresponsiveness (AHR) to bradykinin in mild asthma, whereas AHR to bradykinin is enhanced by inhaled allergens. OBJECTIVE: Hypothesizing that allergen exposure impairs bronchoprotective nitric oxide within the airways, we studied the effect of the inhaled nitric oxide synthase (NOS) inhibitor N(G)-monomethyl-L-arginine (L-NMMA) on AHR to bradykinin before and after allergen challenge in 10 subjects with atopic asthma. METHODS: The study consisted of 3 periods (1 diluent and 2 allergen challenges). AHR to bradykinin (PD(20)BK) was examined before and 48 hours after allergen challenge, both after double-blinded pretreatment with L NMMA or placebo. The accompanying expression of the various NOS isoforms (ecNOS, nNOS, and iNOS) was examined by means of immunohistochemistry in bronchial biopsies obtained after diluent and allergen challenge. RESULTS: After placebo, AHR to BK worsened after allergen challenge in comparison with before allergen challenge (PD(20)BK, 70.8 nmol [range, 6.3-331] and 257 nmol [35.5-2041], respectively; P =.0004). After L-NMMA, preallergen and postallergen PD(20)BK values (50.1 nmol [1.8-200] vs 52.5 nmol [6.9-204]; P =.88) were similarly reduced (P <.01) and not different from the postplacebo/postallergen value (P >.05). After allergen challenge, the intensity of staining in bronchial epithelium decreased for ecNOS (P =.03) and increased for iNOS (P =.009). These changes in immunostaining were correlated with the accompanying worsening in AHR to BK (R(s) = -0.66 and 0.71; P <.04). CONCLUSIONS: These data indicate that allergen exposure in asthma induces increased airway hyperresponsiveness to bradykinin through impaired release of bronchoprotective nitric oxide associated with downregulation of ecNOS. This suggests that new therapeutic strategies towards restoring the balance among the NOS isoforms during asthma exacerbations are warranted. PMID- 11496235 TI - Basophil recruitment and IL-4 production during human allergen-induced late asthma. AB - BACKGROUND: Basophils represent an important source of inflammatory mediators and cytokines after IgE-dependent activation in human beings. OBJECTIVE: To assess the role of basophils in allergic asthma, we measured the number of basophils in the bronchial mucosa and their capacity to express IL-4 mRNA and protein during allergen-induced late asthmatic responses. METHODS: Fiberoptic bronchoscopic bronchial biopsies were obtained at 24 hours from sites of segmental bronchial allergen challenge and control sites in 19 patients with atopic asthma and 6 nonatopic healthy volunteers. Basophil numbers were assessed by immunohistochemistry through use of mAb 2D7. IL-4 mRNA--positive cells were detected through use of in situ hybridization and colocalized to basophils through use of sequential immunohistochemistry/in situ hybridization. IL-4 protein was detected and colocalized to basophils through use of dual immunohistochemistry. RESULTS: After allergen challenge, there was an increase in the median number of 2D7-positive basophils per square millimeter in the bronchial mucosa in patients with asthma (0.9 cells/mm(2) at baseline to 8.8 cells/mm(2) after challenge; P =.002), which also was significantly higher than what was seen in nonasthmatic controls (P =.01). Similarly, IL-4 mRNA--positive cells were increased at 24 hours in patients with asthma (1.4 to 14) in comparison with controls (0 to 0; P =.02). Colocalization studies revealed that 15% and 41% of the basophil population in patients with asthma after allergen challenge expressed, respectively, IL-4 mRNA and protein. Conversely, 19% of IL-4 mRNA-positive cells and 72% of IL-4 protein--positive cells were accounted for by basophils. CONCLUSION: After allergen provocation in sensitive patients with atopic asthma, basophils are recruited to the bronchial mucosa and express IL-4 mRNA and protein, which might contribute to local IgE synthesis and/or tissue eosinophilia or other aspects of allergic inflammation during late responses and ongoing asthma. PMID- 11496236 TI - Possible role of the 4G/5G polymorphism of the plasminogen activator inhibitor 1 gene in the development of asthma. PMID- 11496237 TI - Glucocorticoids induce basophil apoptosis. AB - BACKGROUND: Induction of apoptosis represents an important mechanism by which glucocorticoids (GCCs) exert their anti-inflammatory properties. The effects of GCCs on apoptosis have been determined in various immune cells and found to vary among different cell types. On the other hand, the effects of GCCs on apoptosis of basophils, active participants in allergic inflammation, have remained obscure. OBJECTIVE: The objective of this study was to investigate the effects of GCCs on basophil apoptosis. METHODS: Basophils were highly purified (purity, >97%) by Percoll density gradient centrifugation followed by negative selection. Cell status was determined by their ability to bind annexin V and exclude propidium iodide. DNA fragmentation was determined by flow cytometry. RESULTS: Dexamethasone (DEX) significantly accelerated the decrease in live cells and increased the number of apoptotic cells in a time-dependent fashion. Light microscopy as well as DNA fragmentation assay confirmed the induction of apoptosis by DEX. A half-maximal effect was observed in a DEX concentration range from 10(-9) to 10(-8) mol/L. Sex steroids did not induce basophil apoptosis at all. DEX also induced basophil apoptosis in the presence of low doses of IL-3. CONCLUSION: GCCs exert potent apoptogenic effects on basophils. GCC-mediated apoptogenic effects on basophils might have implications with respect to the mechanism of action of this class of drugs in allergic disorders. PMID- 11496238 TI - Effect of desloratadine and loratadine on rhinovirus-induced intercellular adhesion molecule 1 upregulation and promoter activation in respiratory epithelial cells. AB - BACKGROUND: Rhinoviruses have been recently associated with the majority of asthma exacerbations for which current therapy is inadequate. Intercellular adhesion molecule 1 (ICAM-1) has a central role in airway inflammation in asthma, and it is the receptor for 90% of rhinoviruses. Rhinovirus infection of airway epithelium induces ICAM-1. Desloratadine and loratadine are compounds belonging to the new class of H(1)-receptor blockers. Anti-inflammatory properties of antihistamines have been recently documented, although the underlying molecular mechanisms are not completely defined. OBJECTIVE: We have investigated the effects of desloratadine and loratadine on rhinovirus-induced ICAM-1 expression, mRNA upregulation, and promoter activation. METHODS: Cultured primary bronchial or transformed (A549) respiratory epithelial cells were pretreated with desloratadine and loratadine for 16 hours and infected with rhinovirus type 16 for 8 hours. ICAM-1 surface expression was evaluated with flow cytometry, and ICAM-1 mRNA was evaluated with specific RT-PCR. In A549 cells promoter activation was evaluated with a chloramphenicol acetyltransferase assay, and binding activity of nuclear factor kappa B in nuclear extracts was evaluated with an electrophoretic mobility shift assay. RESULTS: Desloratadine and loratadine (0.1 10 micromol/L) inhibited rhinovirus-induced ICAM-1 upregulation in both primary bronchial or transformed (A549) respiratory epithelial cells. In A549 cells the 2 compounds showed a dose-dependent inhibition with similar efficacy (inhibitory concentration of 50%, 1 micromol/L). Desloratadine and loratadine also inhibited ICAM-1 mRNA induction caused by rhinovirus infection in a dose-dependent manner, and they completely inhibited rhinovirus-induced ICAM-1 promoter activation. Desloratadine also inhibited rhinovirus-induced nuclear factor kappa B activation. Desloratadine and loratadine had no direct effect on rhinovirus infectivity and replication in cultured epithelial cells. CONCLUSION: These effects are unlikely to be mediated by H(1)-receptor antagonism and suggest a novel mechanism of action that may be important for the therapeutic control of virus-induced asthma exacerbations. PMID- 11496239 TI - Inhibition of IgE production in vitro by intact and fragmented intravenous immunoglobulin. AB - BACKGROUND: Intravenous immunoglobulin (IVIG) has been shown to suppress Ig production both in vivo and in vitro. We have previously found that IVIG inhibits IgE synthesis in human tonsillar B cells stimulated with IL-4 and anti-CD40 antibody. OBJECTIVE: The purpose of this study was to further clarify the mechanism behind the inhibition of IgE production by IVIG through comparing the effects of intact whole molecular IVIG and the F(ab')(2) or Fc fragments of IVIG. METHODS: Human B lymphocytes were purified from tonsils. Cell proliferation was measured by means of tritiated thymidine incorporation. IgE was determined by means of ELISA. Cell-cycle analysis was performed by using flow cytometry. RESULTS: Both intact and fractionated IVIG inhibited anti-CD40- and IL-4- stimulated IgE production in a dose-dependent manner. The maximal inhibition was achieved at 67 micromol/L (eg, 10, 6, and 4 mg/mL for intact IVIG, F[ab'](2), and Fc, respectively). The effect of F(ab')(2) was more pronounced than that of Fc at equimolar concentrations. Similarly, both intact and fragmented IVIG dose dependently decreased tritiated thymidine incorporation. F(ab')(2) was also more potent than Fc in this effect. Heat-aggregated IVIG exhibited similar potency to regular IVIG in inhibiting B-cell proliferation. The inhibitory effects of IVIG were unlikely to have been caused by the induction of apoptosis because neither intact nor fractionated IVIG had a significant effect on cell-cycle parameters at the concentrations used. CONCLUSION: These data suggest that both F(ab')(2) and Fc portions contribute to the inhibition of in vitro IgE production by IVIG. The role of the F(ab')(2) portion is more important than that of the Fc portion. PMID- 11496240 TI - Costimulatory molecules in the developing human gastrointestinal tract: a pathway for fetal allergen priming. AB - BACKGROUND: Antigen-specific responses can be detected in umbilical cord blood mononuclear cells. The fetal immune system must therefore attain a level of maturity compatible with the initiation of such responses as well as be exposed to antigen. OBJECTIVE: We sought to assess the expression of costimulatory molecules in fetal gut and the presence of cytokines in amniotic fluid at this time as a preliminary analysis of the suitability of the fetal gut as a site of antigen priming during intrauterine life. METHODS: Human fetal gut was analyzed for cells expressing costimulatory molecules through use of immunohistochemistry. Amniotic fluid was studied by ELISA, for cytokines regulating the nature of the response, and as a source of the common dietary antigen ovalbumin. RESULTS: MHC class II--positive cells were abundant over the period examined (11-24 weeks of gestation), other surface antigens showing spatial and temporal variation in expression. From 11 to 14 weeks of gestation, CD68-positive and CD40-positive cells, like MHC class II--positive cells, were present throughout the lamina propria; few CD3-positive cells (T cells) were observed. With the emergence of lymphoid aggregates (14-16 weeks), CD83-positive cells (dendritic cells) and CD20 positive cells (B cells) could be detected in fetal gut; however, expression was restricted to the lymphoid aggregates. In contrast, MHC class II, CD40, and CD68 continued to be expressed in the lamina propria. CD28-positive cells were also evident from 14 weeks of gestation, occurring throughout the lamina propria and lymphoid aggregates; this corresponded to the increasing numbers of CD3-positive cells. The occasional CD86-positive, CD40L-positive, or CTLA4-positive cell could be seen in or around lymphoid aggregates after 14 weeks of gestation. Lymphoid follicles forming after 16 weeks of gestation contained MHC class II--positive, CD83-positive, CD20-positive, CD40-positive, CD86-positive, CD3-positive, CD28 positive, CD40L-positive, and CTLA4-positive cells. MHC class II--positive, CD40 positive, CD68-positive, CD3-positive, and CD28-positive cells continued to be present in the lamina propria at this time. At all times studied, CD14 was not expressed in the lamina propria or lymphoid follicles. Prostaglandin E(2), TGF beta(1), and IL-10 dominated the amniotic fluid cytokine milieu, and ovalbumin was also detectable in amniotic fluid from 3 of 26 women who had detectable circulating levels. CONCLUSION: Of the costimulatory molecules studied, CD40 was the most abundant. However, both of the ligand families studied (CD40-CD40L and CD86-CD28/CD152) could provide the costimulatory signals required for the initiation of antigenspecific reactivity in the gastrointestinal tract of the human fetus as early as 16 weeks of gestation. The cytokine milieu would favor the development of T(H)2-type reactivity to antigens, such as ovalbumin, that are present at this time. PMID- 11496241 TI - Inhibition of human allergic T-cell responses by IL-10-treated dendritic cells: differences from hydrocortisone-treated dendritic cells. AB - BACKGROUND: Dendritic cells (DCs) are able to induce human allergic T(H)1 responses as well as T(H)2 responses. OBJECTIVE: In this study, we examined the effect of antiinflammatory agents such as IL-10 and hydrocortisone (HC) on the accessory function of DCs and the resulting T-cell response, especially that of T(H)2 cells. METHODS: Naive and memory CD4(+) T cells from atopic donors were stimulated with autologous allergen-pulsed DCs generated from CD14(+) monocytes by culture with GM-CSF/IL-4 and fully matured with IL-1 beta, TNF-alpha, and PGE(2) in the presence or absence of IL-10 or HC. RESULTS: IL-10-treated DCs and, to a lesser extent, HC-treated DCs showed a decreased expression of MHC II molecules, the costimulatory molecule CD86, and the DC-specific marker CD83, as well as a strongly reduced IL-12 secretion. Consequently, T-cell proliferation was reduced after stimulation with IL-10- or HC-treated DCs alike. However, pretreatment of DCs with IL-10 inhibited the production of T(H)1 and T(H)2 cytokines by T cells, whereas HC-treated DCs inhibited production of IFN-gamma but induced an increased release of IL-4 and no change in IL-5. Both effects were long-lasting; cytokine production remained low (which was due not to enhanced apoptosis but to functional hyporesponsiveness) or even increased after restimulation with fully matured DCs. CONCLUSION: These data indicate that IL-10- or HC-treated DCs differ in their ability to influence human allergic T-cell responses. This has major implications for therapeutic strategies aiming at the downregulation of proallergic T(H)2 responses. PMID- 11496242 TI - Preclinical efficacy and safety of mepolizumab (SB-240563), a humanized monoclonal antibody to IL-5, in cynomolgus monkeys. AB - BACKGROUND: Allergic respiratory diseases are characterized by large numbers of eosinophils and their reactive products in airways and blood; these are believed to be involved in progressive airway damage and remodeling. IL-5 is the principal cytokine for eosinophil maturation, differentiation, and survival. Mepolizumab (SB-240563), a humanized monoclonal antibody (mAb) specific for human IL-5, is currently in clinical trials for treatment of asthma. OBJECTIVE: The purpose of this study was to characterize the pharmacologic activity and long-term safety profile of an anti--human IL-5 mAb to support clinical trials in asthmatic patients. METHODS: Naive and Ascaris suum -sensitive cynomolgus monkeys received various dose levels of mepolizumab and were monitored for acute and chronic pharmacologic and toxic responses. RESULTS: To support preclinical safety assessment, cynomolgus monkey IL-5 was cloned, expressed, and characterized. Although monkey IL-5 differs from human IL-5 by 2 amino acids (Ala27Gly and Asn40His), mepolizumab has comparable inhibitory activity against both monkey IL 5 and human IL-5. In A suum--sensitive monkeys, single doses of mepolizumab significantly reduced blood eosinophilia, eosinophil migration into lung airways, and levels of RANTES and IL-6 in lungs for 6 weeks. However, mepolizumab did not affect acute bronchoconstrictive responses to inhaled A suum. In an IL-2--induced eosinophilia model (up to 50% blood eosinophilia), 0.5 mg/kg mepolizumab blocked eosinophilia by >80%. Single-dose and chronic (6 monthly doses) intravenous and subcutaneous toxicity studies in naive monkeys found no target organ toxicity or immunotoxicity up to 300 mg/kg. Monkeys did not generate anti-human IgG antibodies. Monthly mepolizumab doses greater than 5 mg/kg caused an 80% to 100% decrease in blood and bronchoalveolar lavage eosinophils lasting 2 months after dosing, and there was no effect on eosinophil precursors in bone marrow after 6 months of treatment. Eosinophil decreases correlated with mepolizumab plasma concentrations (half-life = 13 days). CONCLUSION: These studies demonstrate that chronic antagonism of IL-5 by mepolizumab in monkeys is safe and has the potential, through long-term reductions in circulating and tissue-resident eosinophils, to be beneficial therapy for chronic inflammatory respiratory diseases. PMID- 11496243 TI - Comparison of CD8(+) T-cell subsets in HIV-infected rapid progressor children versus non--rapid progressor children. AB - BACKGROUND: CD8(+) T-cell subsets have not been adequately described in HIV infected (HIV(+)) children classified with respect to disease progression as rapid-progressors (RPs) and non-rapid progressors (non-RPs). OBJECTIVE: The purpose of this investigation was to determine the distribution of CD8(+) T-cell subsets in HIV(+) children and correlate the findings with degree of immunosuppression and HIV viral burden. METHODS: By means of 3-color flow cytometry, percentages of CD38(+)DR(+), CD28(+), and CD57(+) CD8(+) T-cell subsets were examined in RP (n = 15) and non-RP (n = 36) HIV(+) children and in HIV-exposed but uninfected (n = 11) and HIVunexposed (n = 8) children. The CD8(+) T-cell subsets were correlated with mean CD4(+) T-cell percentages and HIV RNA levels. Analysis of covariance was used for group comparisons for the control of the covariate of age. RESULTS: The HIV-exposed and HIV-unexposed controls were not different from each other in CD8(+) T-cell subset percentages, except that the DR(-)CD38(+)CD8(+) T-cell percentages were higher in the exposed controls than in the unexposed controls. RPs had a higher mean percentage of DR(+)CD38(+)CD8(+) T cells than non-RPs and both control groups, and RPs had higher viremia than non-RPs. CD38(+)CD8(+) T-cell percentages did not correlate with viral burden as it has been seen to do in HIV(+) adults. Percentages of CD28(+)CD8(+) T cells were lower in HIV-infected children than in controls. There was a positive correlation of percentage of CD28(+)CD57(-)CD8(+) T cells with CD4(+) T-cell percentages in each HIV-infected group. CONCLUSION: CD8(+) T cells become activated (dual expression of DR and CD38) and lose CD28, some acquiring CD57, in relation to rapidity of disease progression in pediatric HIV infection. PMID- 11496244 TI - Decline of CD3-positive T-cell counts by 6 months of age is associated with rapid disease progression in HIV-1--infected infants. AB - Because HIV-1 infected infants with rapid progression (RP) of disease might benefit from early and intense antiretroviral therapy, the identification of predictive factors of RP becomes extremely important. Currently, the best predictive factors of RP in HIV-1 infected children are HIV-1 RNA levels and CD4 positive T-cell counts. A decrease in CD3-positive T-cell count has been identified as a predictive factor of AIDS development in HIV-1 infected adults. Our objective was to evaluate decreased number of CD3-positive T-cells as a predictive factor of RP in infants. Peripheral blood lymphocytes from HIV-1 infected infants (up to 6 months of age) were analyzed for an association of lymphocyte subsets with RP, which was defined as the occurrence of AIDS or death before 18 months of age. In infants with RP (n = 32), CD3-positive T-cell counts were 3093 cells/microL at <1 month of age, 3092 cells/microL at 1 to 3 months, and 2062 cells/microL at 3 to 6 months. Non-RP infants (n = 49) maintained their CD3-positive T-cells counts at approximately 4000 cells/microL for at least 6 months of life. CD3-positive and CD4-positive T-cell counts were significantly associated with RP. Our results suggest that a decreased CD3-positive T-cell count may be used to predict RP in HIV-1 infected infants (RR = 2.16, P =.001). PMID- 11496245 TI - Fibronectin and fibrinogen contribute to the enhanced binding of Staphylococcus aureus to atopic skin. AB - BACKGROUND: Staphylococcus aureus colonizes the skin lesions of more than 90% of patients with atopic dermatitis (AD). The mechanism for increased S aureus colonization in AD is unknown. However, the initial event in colonization requires adherence of S aureus to the skin. OBJECTIVE: The purpose of this study was to examine the roles of various bacterial adhesins on S aureus binding to AD skin. METHODS: In an attempt to delineate the mechanism behind this adherence process, an in vitro bacterial binding assay was developed to quantitate the adherence of various S aureus strains to AD, psoriatic, and normal skin sections. S aureus strains used in this study were obtained either from cultures of AD skin lesions or from genetically manipulated strains of S aureus that lacked specific microbial surface components recognizing adhesive matrix molecules (MSCRAMMs)- namely, fibronectin-binding protein (Fnbp), fibrinogen-binding protein (Clf), collagen-binding protein (Cna), and their parent strains. In addition, S aureus strains from patients with AD were pretreated with fibronectin or fibrinogen to block MSCRAMM receptors and interfere with binding. RESULTS: Under all experimental conditions, binding of S aureus was localized primarily to the stratum corneum. Immunocytochemical staining of AD skin sections showed a redistribution of fibronectin to the cornified layer, an observation not seen in normal skin. S aureus binding to uninvolved AD skin was significantly greater than the binding to uninvolved psoriatic skin (P <.0001) and normal skin (P <.0005). The Fnbp-negative S aureus showed a significant reduction in binding to the AD skin (P <.0001) but not to the psoriatic and normal skin. In the AD skin, a significant reduction in the binding of S aureus was also observed in the Clf negative strain (P <.0001) but not in the Cna-negative S aureus. Preincubation of S aureus with either fibronectin or fibrinogen also inhibited bacterial binding to AD skin (P <.0001). CONCLUSION: These data suggest that fibronectin and fibrinogen--but not collagen--play a major role in the enhanced binding of S aureus to the skin of patients with AD. PMID- 11496246 TI - The ascomycin macrolactam pimecrolimus (Elidel, SDZ ASM 981) is a potent inhibitor of mediator release from human dermal mast cells and peripheral blood basophils. AB - BACKGROUND: The ascomycin macrolactam pimecrolimus (Elidel, SDZ ASM 981) has recently been developed as a novel and cell-selective inhibitor of inflammatory cytokine secretion; it has fewer adverse effects than currently available drugs. OBJECTIVE: In this study, we investigated the capacity of pimecrolimus to directly inhibit in vitro mediator release from human skin mast cells and basophils. METHODS: Purified cutaneous mast cells or basophil-containing peripheral blood leukocytes were obtained from healthy human donors and preincubated with pimecrolimus (0.1 nmol/L to 1 micromol/L) in the absence or presence of its specific antagonist (rapamycin), cyclosporin A (100 nmol/L to 1 micromol/L), or dexamethasone (1 micromol/L) and then stimulated with anti-IgE or with calcium ionophore A23187 plus phorbol myristate acetate. Cell supernatants were kept for analysis of histamine, tryptase, LTC4, and TNF-alpha. RESULTS: Pimecrolimus caused a strong and dose-dependent inhibition of anti-IgE--induced release of histamine from mast cells and basophils (maximally 73% and 82%, respectively, at 500 nmol/L pimecrolimus) and of mast cell tryptase (maximally 75%) and a less pronounced inhibition of LTC4 (maximally 32%) and of calcium ionophore plus phorbol myristate acetate--induced mast cell TNF-alpha release (90% maximum at 100 nmol/L pimecrolimus). In contrast, inhibition achieved during mast cell histamine release was maximally 60% with cyclosporin A and only 28% with dexamethasone. CONCLUSION: These data demonstrate a marked inhibitory capacity of pimecrolimus on mediator release from human mast cells and basophils with a potency exceeding that of cyclosporin A and dexamethasone. Pimecrolimus might thus be expected to be effective in the treatment of mast cell-- and basophil-dependent diseases. PMID- 11496247 TI - Atopic dermatitis is associated with a low-producer transforming growth factor beta(1) cytokine genotype. AB - BACKGROUND: Atopic dermatitis (AD) is associated with hyperresponsiveness of lymphocytes to allergens. In acute AD only T(H)2-type lymphocytes are activated, whereas in more chronic forms of AD, the activity of both T(H)1- and T(H)2-type lymphocytes increases. IL-10 and transforming growth factor beta(1) (TGF-beta(1)) are immunosuppressive cytokines that inhibit the activity of both T(H) cell types in human subjects. OBJECTIVE: The aim of this study was to determine whether children with moderately severe chronic AD had IL10 or TGFB1 genotypes known to be associated with low cytokine production. METHODS: Using amplification refractory mutation screening PCR, we examined TGFB1 and IL10 gene polymorphisms, which are known to affect cytokine production, in 68 children with moderately severe AD and in 50 nonatopic children. RESULTS: The odds ratio of children with AD having a low TGFB1 producer genotype was 4.8 (95% CI, 2.4--9.7) compared with the control subjects (P <.0001). There were no differences in the frequency of IL10 gene polymorphisms between groups. CONCLUSION: TGFB1 genotype may partly explain the strong genetic predisposition to AD. PMID- 11496248 TI - The -590C/T and -34C/T interleukin-4 promoter polymorphisms are not associated with atopic eczema in childhood. AB - Susceptibility to the development of asthma and other atopic diseases is known to have a genetic component. To date, several studies have linked chromosome 5q31 to asthma and atopy in human beings. This region harbors a cluster of cytokine and growth factor genes, IL-4 presenting as a prime atopy candidate gene, inasmuch as it plays a pivotal role in the atopy pathway. Our approach was to identify polymorphisms within the promoter regions of IL-4 and test their association with atopic eczema. Polymorphisms were typed in a cohort of 76 small nuclear families and 25 triads with childhood atopic eczema. The genotypes were used to test for linkage in the presence of association with atopic eczema. A new polymorphism, 34C/T, was identified and studied with a known polymorphism, -590C/T. On its own, each polymorphism showed no association with atopic eczema. The 2 polymorphisms were used to generate haplotypes, and a significant result was found for the 590C/-34C haplotype. However, after Bonferroni correction for multiple testing, the association became nonsignificant. Neither polymorphism predisposes to early onset atopic eczema by itself, but suggestive linkage was found for the -590C/ 34C haplotype in this study. PMID- 11496249 TI - Elimination of mouse allergens in the working environment: assessment of individually ventilated cage systems and ventilated cabinets in the containment of mouse allergens. AB - BACKGROUND: Laboratory animal allergy is an important occupational disease that is preventable by reduction of exposure to mammalian allergens. OBJECTIVE: The purpose of this investigation was to assess the efficacy of safety equipment in containing mouse urinary protein (MUP)--specifically, individually ventilated cage (IVC) systems and class I--type and class II ventilated cabinets. METHODS: Six IVC systems (which are used to house rodents) were operated at positive and negative pressure. Air samples (2 L/min) were collected at the cage front, cage back, air inlet, and air outlet, and the MUP was quantified by immunoassay. The background MUP of the IVC study room was compared with that of rooms where mice were housed conventionally or in isolators. Class I--type cabinets (n = 2) were tested during the disposal of soiled litter. Air samples were positioned on and behind the operator and inside the cabinet (n = 2). Personal samples were collected while scientific procedures were performed in a class II cabinet and in the open. RESULTS: All of the IVC systems contained MUP effectively (>250-fold) when operated at negative pressure. At positive pressure, the "unsealed" IVC racks leaked MUP (1- to 25-fold reduction) but the "sealed" IVC system did not. Class I--type cabinets reduced (10-fold) but did not eliminate exposure during "cleaning out." No MUP was detected when procedures were performed in class II cabinets (protection factor, >10-fold). CONCLUSION: Safety equipment can substantially reduce exposure to mouse allergen. Allergen exposure in holding rooms will be minimized if mice are housed in IVC systems operated at negative pressure or in "sealed" IVC systems at positive pressure. Respiratory protection should be used whenever "unsealed" IVC systems are operated at positive pressure or during "cleaning out" in class I--type cabinets. PMID- 11496250 TI - Occupational IgE-mediated allergy after exposure to lupine seed flour. AB - The ingestion of lupine seed flour (LSF) has been reported as a cause of allergic reactions, particularly in patients sensitized to peanut, but there is little evidence of its allergenic potential after inhalation. We sought to evaluate the clinical and immunologic reactivity to lupine in employees working with this seed flour. An occupational history was obtained in 7 subjects (median age, 35 years) working with LSF at an agricultural research center. Three subjects (1, 6, and 7) reported work-related allergy symptoms immediately after being exposed to lupine. Skin prick test results with LSF extract were positive in these 3 patients with work-related symptoms. Moreover, lupine-specific IgE antibodies were detected in subjects 6 and 7. In subject 6, the controlled exposure to LSF elicited immediate naso-ocular symptoms without changes in FEV(1). In subject 7, a bronchial provocation with LSF extract elicited an immediate fall (25%) in FEV(1). Double blinded, placebo-controlled LSF oral challenge results were positive in subjects 6 and 7. Immunologic reactivity to other legumes was detected in subjects 6 and 7, but specific inhalation testing and oral challenge results were negative. Thus, the inhalation of lupine flour could be an important cause of allergic sensitization in exposed workers and might give rise to occupational asthma and food allergy. PMID- 11496251 TI - Severe anaphylaxis induced by latex as a contaminant of plastic balls in play pits. AB - BACKGROUND: Latex causes anaphylaxis in specific contexts among children. We present 2 cases that show that severe reactions may occur in everyday circumstances with latex as a contaminant. OBJECTIVE: Because 2 cases of severe reactions to latex suggested similar circumstances of exposure, we investigated the immediate environment in which episodes occurred. METHODS: A 5-year-old girl presented to our casualty department with anaphylaxis after playing in a ball pit filled with approximately 10-cm diameter plastic balls in an American-style fast food outlet. Two months later, a 9-year-old boy had severe anaphylaxis followed by an asthma attack with loss of consciousness while playing in the playpen of a different outlet belonging to the same company. Latex sensitization was confirmed in both cases by means of skin prick testing, latex glove skin prick testing, and 1-glove finger testing. Immunoblotting of elutions from a ball, the natural rubber latex foam pit lining, and its polyvinyl chloride sheet were performed. RESULTS: In the girl's immunoblot high levels of IgE specific to Hev b 4, Hev b 7, and Hev b 2 were found. The boy's immunoblot showed positivity to Hev b 7. The polyvinyl chloride ball sample showed a high concentration of specific Hevea species allergen similar to that of the foam layer sample. CONCLUSION: Severe anaphylaxis can result from contact with latex proteins as a contaminant, rather than as a component, of play area ball pits and therefore outside the reported settings. Emergency health care workers should be aware of this kind of risk. A latex-reduced environment might prevent potentially severe reactions in young customers of fast-food outlets. PMID- 11496252 TI - Carrot allergy: double-blinded, placebo-controlled food challenge and identification of allergens. AB - BACKGROUND: Allergic reactions to carrot affect up to 25% of food-allergic subjects. Clinical manifestations of carrot allergy and IgE responses to carrot proteins, however, have never been studied in subjects with carrot allergy confirmed by means of double-blinded, placebo-controlled food challenge (DBPCFC). OBJECTIVE: The purposes of this investigation were to confirm clinically relevant sensitizations to carrot by means of DBPCFC, to validate current diagnostic methods, and to identify IgE-reactive carrot proteins in patients with true allergy. METHODS: DBPCFCs were performed in 26 subjects with histories of allergic reactions to carrot. Patients underwent skin prick tests with carrot extract, fresh carrot, and various pollen extracts. Specific IgE to carrot, celery, birch, and mugwort pollen and to rBet v 1, rBet v 2, and rBet v 6 were measured through use of the CAP method. Carrot allergens were identified by means of immunoblotting and blotting inhibition. RESULTS: Twenty of 26 patients had positive DBPCFC results. The sensitivity of the determination of carrot-specific IgE antibodies through use of the CAP method (> or =0.7 kU/L) was 90%, the sensitivity for skin prick testing with commercial extracts was 26%, and the sensitivity for prick-to-prick tests with raw carrot was 100%. The Bet v 1- related major carrot allergen Dau c 1 was recognized by IgE from 85% of patients; 45% were sensitized to cross-reactive carbohydrate determinants and 20% to carrot profilin. In 1 subject, a Bet v 6--related carrot allergen was recognized. In 4 patients, IgE binding to Dau c 1 was not inhibited or was weakly inhibited by rBet v 1 or birch pollen extract. CONCLUSION: This study confirmed the allergenicity of carrot by means of DBPCFC. DBPCFC-positive patients had exclusively specific IgE antibodies to birch pollen--related carrot allergens, Dau c 1 being the major allergen. The lack of inhibition of IgE binding to Dau c 1 by birch allergens in a subgroup of patients might indicate an secondary immune response to new epitopes on the food allergen that are not cross-reactive with Bet v 1. PMID- 11496253 TI - Effective allergen avoidance reduces residual volume and sputum eosinophils in children with asthma. PMID- 11496254 TI - Improvement in atopic dermatitis in infants with the introduction of an elemental formula. PMID- 11496255 TI - A case of onion allergy. PMID- 11496256 TI - T-cell apoptosis in ICF syndrome. PMID- 11496259 TI - Mistreatment of elders by family members. PMID- 11496260 TI - Medication management in the community: some considerations about compliance. PMID- 11496261 TI - Arthritis: taking control. PMID- 11496262 TI - Home oxygen supplies threatened by retesting proposal. PMID- 11496263 TI - Just the facts, please: a guide to effective documentation. AB - Although most health care professionals would wish it otherwise, documentation will never be eliminated completely. It can be ignored and complained about, but documentation will always be there to remind us, "If it wasn't documented, it wasn't done." Home care providers need to be cognizant of the imperative for effective documentation. Corporate compliance initiatives, fraud and abuse issues, and changes in reimbursement mechanisms make documentation more important than ever. Home care providers must create an environment conducive to effective documentation practices. PMID- 11496264 TI - Palliative care: long-term solution for long-term care. Part 2. Types of pain: a review. PMID- 11496265 TI - Lessons learned from the floods of Hurricane Floyd: a disaster of biblical proportions. PMID- 11496267 TI - Identification and implications of transgraft microleaks after endovascular repair of aortic aneurysms. AB - PURPOSE: The purpose of this report is to describe an interesting cause of endoleak and detail-specific techniques for identifying small transgraft defects, which we have termed microleaks. METHODS: Four patients underwent endovascular repair of abdominal aortic aneurysms with modular nitinol/polyester endoprostheses and were studied after 6 to 30 months. All patients were enrolled in standard follow-up radiographic surveillance protocols. RESULTS: Three of the four abdominal aortic aneurysms continued to expand after endograft repair. Standard computed tomography imaging with precontrast, dynamic contrast, and delayed imaging frequently identifies endoleak, although it fails to precisely identify microleaks as the source. Color flow duplex ultrasound scan was performed on three patients and perigraft "jets," small areas of color flow adjacent to the endograft, were identified in all. Microleaks were identified in one patient who underwent digital subtraction arteriography with directed efforts to completely opacify the prosthesis lumen and multiple oblique projections. In another patient, contrast arteriography with balloon occlusion of the distal endograft clearly depicted midgraft microleaks that might otherwise be mistaken for graft porosity or cuff junction endoleaks. No microleaks were diagnosed on angiograms when these directed efforts were not performed. Aneurysm exploration before aortic clamping provided conclusive determination of the presence of blood flow through the wall of the endoprosthesis in two patients. CONCLUSIONS: Microleaks occur up to 2.5 years after endovascular repair of aortic aneurysms. Although computed tomography demonstrates the presence of an endoleak in these patients, the exact site of origin usually remains obscure. Doppler ultrasound scan and directed arteriography appear to be of greater utility for identifying the presence and location of microleaks. Balloon occlusion arteriography and aneurysm exploration without arterial clamping provide definitive evidence of microleaks. Although the clinical significance of microleaks remains unclear, long-term monitoring of patients is imperative to diagnose and treat these and other modes of endograft failure before they progress to aneurysm rupture. PMID- 11496268 TI - Serious complications that require surgical interventions after endoluminal stent graft placement for the treatment of infrarenal aortic aneurysms. AB - BACKGROUND: Endoluminal stent-graft placement for the treatment of infrarenal abdominal aortic aneurysms has gained widespread acceptance because it is associated with lower peri-interventional morbidity than conventional transabdominal surgery. In this study the long-term morbidity of the procedure was evaluated. METHODS AND RESULTS: Between September 1994 and December 1998, 150 patients (age, 69.6 +/- 8.49 years; 142 men, 8 women) with abdominal aortic aneurysms were treated with the placement of an intravascular nitinol stent-graft (Stentor [55] and Vanguard-System [95]); there were eight tubular grafts and 142 bifurcated grafts. Initial placement of the stent-graft was successful in 144 patients. Mean follow-up was 49 +/- 25 months. In 13.3% of stent-graft placements we encountered the following complications: 4 cases of migration or dislocation of the prosthesis (30.5 +/- 7.4 months after placement); 2 ruptures of the aorta (26.7 and 15.0 months after placement); 3 recurrent thromboses of the stent-graft (25.5 +/- 5.3 months after placement); 3 endoleaks (27.5 +/- 15.7 months after placement); and 5 infections of the prosthesis (26.6 +/- 16.5 months after placement). There was no correlation between the complications and the type of stent used. All of these patients were treated by surgical replacement of the prosthesis with a Dacron graft. CONCLUSIONS: (1) The results suggest that most complications are due to a continuation of the disease process leading to loosening of the prosthesis. (2) Explantation of the prosthesis and surgical repair is feasible but bears additional risks. (3) Because the onset of reperfusion of the excluded aneurysm cannot be predicted, all patients with infrarenal aortic stent-grafts require frequent computer tomographic follow-up. (4) Finally, the results call for further improvements in the design of the stent graft. PMID- 11496269 TI - Intentional internal iliac artery occlusion to facilitate endovascular repair of aortoiliac aneurysms. AB - PURPOSE: The safety of intentional occlusion of patent internal iliac arteries (IIAs) to facilitate the endovascular repair of aortoiliac artery aneurysms (abdominal aortic aneurysms [AAAs] and iliac aneurysms [IAs]) was evaluated. METHODS: We analyzed the techniques and clinical sequelae of selective occlusion of one or both IIAs in 103 patients and correlated these findings with the results of preoperative angiograms to identify vascular anatomy that may predict postoperative pelvic ischemia. To quantify the clinical presentation of pelvic ischemia, we developed these criteria: class 0, no symptoms; class I, nonlimiting claudication with exercise; class II, new onset impotence, with or without moderate to severe buttock pain, leading to physical limitation with exercise; class III, buttock rest pain, colonic ischemia, or both. IIA occlusion was achieved in 100% of the patients by means of either catheter-directed embolization or orificial coverage with a stent-graft. No patient in this study had angiographic evidence of significant visceral occlusive disease before the procedure. Sixty-four patients had isolated AAAs, 23 patients had AAAs and IAs, and 16 patients had isolated IAs. Ninety-two patients had one IIA selectively occluded, and 11 patients had both IIAs selectively occluded. RESULTS: After IIA occlusion, 12 patients were categorized in class I, 9 patients were categorized in class II, and 1 patient was categorized in class III, for a total of 22 patients (21%) with pelvic ischemia. Sixteen (17%) of 92 patients had unilateral IIA occlusions, and six (17%) of 11 patients had bilateral IIA occlusions. Five patients in class I improved and had no symptoms within 1 year, and one patient in class II was downgraded to class I because of improved symptoms. Two unique preoperative angiographic findings were identified in the remaining 16 patients (16%) with chronic pelvic claudication: (1) stenosis of the remaining IIA origin (> 70%) with nonopacification of more than three of the six IIA branches (63%); and (2) small caliber, diseased or absent medial and lateral femoral circumflex arteries ipsilateral to the side of the IIA occlusion (25%). One patient with class III ischemia died of cardiovascular collapse associated with colon infarction caused by either acute ischemia or particulate embolization. CONCLUSION: The incidence of pelvic ischemia after IIA occlusion is 20% immediately after endovascular aortoiliac aneurysm repair. A total of 25% of patients had no symptoms within 1 year. Two preoperative radiologic findings may help identify patients who are at risk for pelvic ischemia: stenosis of the patent IIA and disease deep femoral ascending branches ipsilateral to the occluded IIA. The risk of colon ischemia appears to be small after selective IIA occlusion to facilitate endovascular AAA repair. PMID- 11496270 TI - Indicators of infection with Chlamydia pneumoniae are associated with expansion of abdominal aortic aneurysms. AB - PURPOSE: Chlamydia Pneumoniae has been shown to be associated with atherosclerosis, myocardial infarction, and abdominal aortic aneurysms (AAAs). The possible association between AAA expansion and C pneumoniae infection was therefore assessed. METHODS: Blood samples were taken from patients with an AAA that was considered for surgical repair after having been diagnosed by means of the Chichester aneurysm screening program (UK) as having an initially infrarenal aortic diameter of 3.0 to 5.9 cm. The patients were examined prospectively for as long as 11.5 years (mean, 4.1 years) with ultrasound scanning. Of 110 patients considered for surgery, 90 men and 10 women had blood samples taken. Their IgG and IgA antibodies against C pneumoniae were measured by means of a microimmunofluorescence test. Unpaired t tests, multiple linear regression analyses, and logistic regression analyses were used for statistical analysis. RESULTS: A total of 44% (95% CI, 31%-55%) of the men with an AAA had an IgA titer of 64 or more, an IgG titer of 128 or more, or both, compared with 10% of the women with an AAA (OR = 7.2; 95% CI, 1.05-160.8). A titer of IgG of 128 or more was significantly associated with higher expansion (5.3 vs 2.6 mm per year), even after adjustment for initial AAA size and age. A significant positive correlation between both IgA and IgG titers and mean annual expansion was observed (r = 0.28; 95% CI, 0.05-0.49; and r = 0.45; 95% CI, 0.24-0.62, respectively), persisting after adjusting for initial AAA size and age. An IgG titer of 128 or more was present significantly more often in cases with an expansion greater than 1 cm annually (adjusted OR = 12.6; 95% CI, 1.37-293). CONCLUSION: A high proportion of men with an AAA has signs of infection with C pneumoniae. The progression of their AAAs was positively correlated with the presence of indicators of C pneumoniae infection. PMID- 11496271 TI - Laparoscopy-assisted aneurysm resection as a minimal invasive alternative in patients unsuitable for endovascular surgery. AB - OBJECTIVE: So far, endovascular surgery has been the only minimal invasive way to treat patients with abdominal aortic aneurysms (AAAs). With hand-assisted laparoscopic surgery (HALS), laparoscopic transperitoneal endoaneurysm repair can be performed through a 6-cm mini-incision only. We wanted to evaluate whether this laparoscopic technique can be offered as a minimal invasive alternative in patients unsuitable for endovascular AAA repair. MATERIAL AND METHODS: Forty patients were referred for endovascular AAA repair. Three patients had to be excluded from the study. Endovascular AAA exclusion was finally performed in 13 patients. Laparoscopic AAA resection was performed in 24 patients. Hand-assisted laparoscopic surgery with transperitoneal access and endoaneurysm repair was accomplished in all patients unsuitable for an endovascular procedure. The outcome after endovascular repair was compared with the outcome of patients who underwent laparoscopy. RESULTS: In the laparoscopic group, conversion to an open procedure was necessary in one case. One patient in this group died (4.1%) postoperatively. There were four complications in each group. In the endovascular group we had one endoleak type II and one graft thrombosis, which required a reoperation. After endovascular treatment, patients were transferred significantly less frequently to the intensive care unit, and they could resume oral feeding earlier. Mobilization and postoperative hospital stay did not differ significantly between the groups. CONCLUSION: Laparoscopic AAA resection with the use of the technique described can be routinely offered to patients unsuitable for endovascular AAA exclusion with excellent long-term results similar to open surgery. A controlled study is clearly indicated to evaluate the role of laparoscopic techniques in aneurysm surgery. PMID- 11496272 TI - Aneurysms of the innominate artery: surgical treatment of 27 patients. AB - PURPOSE: Aneurysms of the innominate artery (AIA) are widely considered to be a rare entity. We describe our experience with AIAs in the last three decades. METHODS: From October 1973 to October 2000, we operated on 27 patients with an AIA. The underlying cause of aneurysm was Takayasu's disease in 7 patients, degenerative disease in 6 patients, syphilis in 5 patients, chronic dissection in 3 patients, trauma in 2 patients, infection in 2 patients, a postoperative complication in 1 patient, and Marfan syndrome in 1 patient. AIA was associated with an aortic aneurysm in 17 patients. Fourteen patients had no symptoms. The remaining patients had symptoms, with thromboembolic complications in 7 patients, pain without rupture in 3 patients, and a ruptured aneurysm in 3 patients. In two patients at high risk for surgery who had a small AIA with embolic complications, a cervical approach was used as a means of performing distal exclusion and crossover bypass. In the remaining 25 patients, a midline sternotomy was used. One patient with a ruptured AIA exsanguinated during sternotomy. Ten patients underwent a prosthetic replacement of the ascending aorta and/or aortic arch with a separate prosthetic branch to the innominate artery (IA). Thirteen patients underwent ascending aorta-to-IA prosthetic bypass in association with lateral suture (8 patients) or prosthetic patching (5 patients) of the aorta. One patient with an infected aneurysm was treated by means of resection of the aneurysm, proximal ligation of the IA, and transposition of the right into the left common carotid artery. Cardiopulmonary bypass with deep hypothermic circulatory arrest was used in 10 patients. RESULTS: Three perioperative deaths occurred (2 of 4 in association with emergency treatment and 1 of 23 with elective treatment). Respiratory complications requiring prolonged artificial ventilation developed in five patients. Two patients had transient worsening of preoperative neurologic deficits. Late results, with a mean follow-up of 85 months, were good. CONCLUSION: The etiology and presentation of AIAs are variable. Surgical management with current cardiovascular techniques achieves excellent results. PMID- 11496273 TI - Influence of diagnostic criteria on the prevalence of abdominal aortic aneurysm. AB - PURPOSE: We studied the prevalence of abdominal aortic aneurysm (AAA) in a population with high incidences of cardiovascular diseases and analyzed how the prevalence varies according to methodology and criteria. METHODS: All men and women aged 65 to 75 years who lived in the Norsjo municipality in northern Sweden were invited to undergo an ultrasound scanning (US) examination. Those with an aortic diameter of 28 mm or more or with poor visibility on US were examined with computed tomography scanning (CT). Various recommended AAA definitions, two diagnostic methods (US and CT), and two diameters (maximum and anteroposterior) were analyzed. RESULTS: Of 555 people invited to participate in the study, 504 accepted (248 men and 256 women; 91%). Eight subjects had undergone surgery for an AAA. Ninety-two subjects underwent CT. The mean maximum infrarenal aortic diameter was 24.6 mm (by means of US). Depending on diagnostic criteria, the AAA prevalence was 3.6% to 16.9% in men and 0.8% to 9.4% in women. Depending on which previous study was used as a comparison and the definition of AAA and diagnostic technique used, the prevalence in this study was 1.3 to 4.0 times higher for men and 2.0 to 5.8 times higher for women. CONCLUSION: In a region in which residents have a high risk for cardiovascular disease, we found the highest prevalence of AAA ever reported within a population. The prevalence highly depends on methodology and diagnostic criteria, with a 10-fold variation. Detailed defined criteria are necessary to permit comparisons between studies: the number of individuals who have undergone surgery for AAA and whether they are included, the prevalence in 5- and 10-year age intervals, attendance rate, visibility, which diameter(s) is measured, and the prevalences with as many as possible of the four described definitions of AAA. The etiology of the high prevalence of AAA in this population needs to be investigated further. PMID- 11496274 TI - Late recurrent saphenofemoral junction reflux after ligation and stripping of the greater saphenous vein. AB - OBJECTIVE: This study was done to determine the long-term incidence of refluxing epifascial-to-deep vein reconnections in the area of the former saphenofemoral junction after ligation of the true junction, along with all proximal tributaries, and resection of the greater saphenous vein. PATIENTS AND METHODS: A total of 125 limbs in 77 patients, representing 66% of 117 survivors among 602 patients who underwent operation between 1960 and 1967, were evaluated clinically and with duplex sonography for possible superficial-to-deep vein reconnections and clinical recurrence of thigh varicosities at a mean follow-up of 34 years. RESULTS: Clinical examination suggested saphenofemoral recurrence in 59 limbs (47%). In 11 instances these were actually varices associated with isolated superficial system reflux or reflux originating from a distally located perforating vein. Color-coded duplex ultrasonography demonstrated saphenofemoral reflux in 75 limbs (60%), versus the 48 identified on clinical examination (P <.001), and documented that the junction ligation had not been performed incorrectly by absence of the terminal valve or any patent proximal saphenous remnant. The reflux originated at the site of the ligated saphenofemoral junction in 53 limbs (71%) and from a nearby circumjunctional deep vein in the other 22 (29%). Of the real junctional recurrences, 22 appeared as a tangled cluster, and 31 involved a single-lumen varix. Only 27 recurrences were sufficiently symptomatic to warrant consideration of additional treatment; 25 of these were clinically evident, single-lumen, true junctional recurrences. CONCLUSIONS: This 34-year clinical follow-up study shows a 60% incidence of junctional and circumjunctional reconnections after ligation of the true saphenofemoral junction and its related tributaries. Color-coded duplex sonography is a necessary concomitant to clinical examination, detecting more recurrences and defining the pathologic anatomy to direct clinically indicated additional treatments. PMID- 11496275 TI - Fascial relationships of the short saphenous vein. AB - OBJECTIVES: The purpose of this study was to define the relationships between the short saphenous vein (SSV) and the fasciae of the leg, including the muscular fascia (MF) and the membranous layer (ML) of the subcutaneous tissue. METHODS: Fascial relationships of the SSV were evaluated by means of dissection in 30 cadaveric limbs and by means of duplex sonography in 270 healthy limbs from living subjects. RESULTS: All along the leg, the SSV courses in a flat compartment delimited by the MF and the ML. Neither results from dissection nor results from sonographic examination demonstrated piercing of the MF by the SSV. A hyperechoic lamina similar to a ligament connects the SSV to the fasciae by which it is encased. An SSV tributary and collateral vessels course out of this space and are devoid of any fascial wrapping. CONCLUSIONS: The SSV does not correspond to the classical description of a "superficial" vein. In fact, from the anatomical point of view, the SSV is an interfascial vein, because it is encased by two connective fasciae, just like the greater saphenous vein. Fascial relationships of the SSV suggest that muscular contraction potentially influences the caliber and hemodynamics of the SSV. In addition, the ML is arranged as a sort of mechanical shield that could counteract dilative pathologic conditions in varicose limbs. PMID- 11496276 TI - Underlying factors influencing the development of the post-thrombotic limb. AB - PURPOSE: This study was designed to determine whether patients having underlying venous disease in their contralateral limbs indicates a more severe long-term clinical outcome in the ipsilateral limb after a deep vein thrombosis (DVT) and to determine what other factors may influence the long-term outcome. METHODS: An acute DVT was initially diagnosed by means of duplex ultrasound scanning. Follow up clinical examinations and bilateral duplex reflux studies were performed for a mean period as long as 3 years. The patients were divided into two groups: group I, those with no history of a contralateral DVT, and group II, those with a history of a contralateral DVT. The patients were classified at their final examination according to the Clinical, Etiology, Anatomic, Pathophysiologic (CEAP) classification, and the ipsilateral and contralateral limbs were compared. Predisposing factors were compared with the final clinical outcome. RESULTS: Sixty-three patients were monitored in a mean follow-up period of 3 years. There was a significant difference in the incidence of symptoms between the ipsilateral limbs (P <.01) and the contralateral limbs (P <.001) for both groups. There was no significant difference between the incidence of superficial reflux between the ipsilateral and contralateral limbs, but the deep venous system and perforator veins were involved more often in the ipsilateral limbs. In group I, only six patients (10%) had no evidence of venous dysfunction (CEAP = 0) in their ipsilateral or contralateral limbs at the time of the final examination, and all patients had reversible risk factors. Of patients who had a mild clinical outcome (CEAP score, 1 to 3), 64% had a healthy contralateral limb, and the remaining 36% had mild to moderate disease. Eighty percent of patients with the most severe clinical outcome (CEAP > 3) had mild to moderate venous disease in their contralateral limb and had nonreversible risk factors. CONCLUSION: There are a significant number of patients with an acute DVT who had an underlying venous disease in the uninvolved contralateral limb. An ipsilateral post-thrombotic limb is more likely to develop in patients with primary venous reflux after an acute DVT. The level of venous dysfunction in the contralateral limb is an indication of the severity of disease developing in the ipsilateral limb. The initial risk factors of the patients have an influence on the final clinical outcome. PMID- 11496277 TI - Cost-effectiveness of oral anticoagulants versus aspirin in patients after infrainguinal bypass grafting surgery. AB - PURPOSE: Several antithrombotic therapies are available for the treatment of patients with peripheral vascular diseases. It is unknown how quality of life and costs of treatment are influenced by different therapies. This study assessed the cost-effectiveness of oral anticoagulants versus aspirin in patients after infrainguinal bypass grafting surgery. METHODS: Clinical outcome events and event free survival were collected from 2650 patients in 77 centers who participated in the Dutch Bypass Oral anticoagulants or Aspirin trial. Approximately half the patients had critical ischemia; 60% received vein grafts, and 20% had femorocrural bypass grafts. A model that was primarily driven by clinical outcome events was used as a means of determining quality of life (EuroQol EQ-5D) and costs for each patient. The main outcome measure was the incremental health care costs in relation to the additional number of quality-adjusted life years and the additional number of event-free years. RESULTS: The mean costs during the 21 months of follow-up were epsilon 6875 per patient in the oral anticoagulants group versus epsilon 7072 in the aspirin group (difference, 197; 95% CI, -746 to 343). The event-free survival was 1.10 years in the group treated with oral anticoagulants versus 1.09 years in the group treated with aspirin (difference, 0.01; 95% CI, -0.07 to 0.08), whereas the corresponding quality-adjusted life years were 1.06 and 1.05, respectively (difference, 0.01; 95% CI, -0.03 to 0.06). CONCLUSION: Health care costs, event-free survival, and quality-adjusted life years in patients after infrainguinal bypass surgery were not different in patients treated with aspirin and patients treated with oral anticoagulants. The extra costs of monitoring patients treated with oral anticoagulants were limited and play no role in the decision for treatment. PMID- 11496278 TI - Influence of poststenotic collateral pressure on blood flow velocities within high-grade carotid artery stenosis: differences between morphologic and functional measurements. AB - PURPOSE: The driving force for blood flow through a high-grade stenosis in the internal carotid artery can be expressed as the pressure gradient over the stenosis itself, which, however, might be reduced by the back pressure exerted by distal collateral vessels. Theoretically the maximum blood flow velocity as a measure of the functional grade of obstruction may therefore be lower than what is expected from morphologic gradations of the stenosis. This study was designed to test prospectively the influence of intracranial collateral vessels on blood flow velocities within high-grade internal carotid artery stenoses. PATIENTS AND METHODS: Forty-five consecutive patients (age 66 +/- 11) with high-grade internal carotid artery stenoses were investigated before and during carotid endarterectomy. The preoperative investigations included duplex ultrasound scanning of the neck vessels, transcranial Doppler scanning for assessment of collateral flow to the middle cerebral artery and angiography. Carotid endarterectomy was performed with patients under deep general anesthesia without a shunt. Systolic and diastolic internal carotid artery blood pressures were measured before and during intraoperative cross-clamping (ie, stump pressure) of the carotid arteries. RESULTS: Within high-grade internal carotid artery stenoses, maximum systolic and end-diastolic blood flow velocities showed a significant inverse correlation to the corresponding systolic and diastolic stump blood internal carotid artery blood pressures. All patients with spontaneous collateral flow to the ipsilateral anterior part of the circle of Willis were divided into a group with relatively high and another one with low end-diastolic blood flow velocities. The stump pressure was significantly lower in patients with high end-diastolic blood flow velocities in spite of the fact that the mean angiographic grade of stenosis did not differ significantly between the groups. CONCLUSIONS: Flow velocities within a high-grade internal carotid artery stenosis are inversely dependent on the stump pressure, that is the poststenotic collateral perfusion pressure. This should be taken into consideration in case of discrepancies between angiography and ultrasound outcome. PMID- 11496279 TI - Intraoperative flow predicts the development of stenosis in infrainguinal vein grafts. AB - OBJECTIVE: There are data to suggest that the development of myointimal hyperplasia is affected by long-term alterations in blood flow. However, the clinical relevance of these findings has not been demonstrated. METHODS: In this retrospective clinical study, intraoperative volume flow measurement with transit time flowmeter was performed in 257 infrainguinal vein grafts carried out in 241 patients. The patients were enrolled in an intensive duplex scanning-based surveillance program. The relationship between the intraoperative graft flow and subsequent occlusion or development of stenosis was evaluated and controlled for other pertinent risk factors. RESULTS: The median follow-up time was 13.6 months. A graft stenosis was found in 58 grafts. The mean graft flow for event-free grafts was 98 mL/min, which was significantly higher compared with 78 mL/min for stenosed or 69 mL/min for occluded grafts. The patients were divided into four groups according to quartiles of the sample distribution of graft flow measurements. The respective 2-year primary and assisted primary patency rates in the lowest to the highest graft flow groups were 39%, 49%, 47%, and 72% (P =.003) and 55%, 67%, 71%, and 84% (P =.01). Analogous significant differences were observed for maximal flow capacity measurements. Female sex (P =.009) and low graft flow in maximal flow capacity measurements (P =.003) were independent predictors of stenosis development in the multiple regression model. CONCLUSION: Intraoperative graft volume flow is a predictor of bypass occlusion after infrainguinal bypass. In addition, this study verifies an association between the development of clinically evident graft stenoses and low graft flow. PMID- 11496280 TI - Day-to-day variability of transcutaneous oxygen tension in patients with diabetes mellitus and peripheral arterial occlusive disease. AB - PURPOSE: We evaluated the day-to-day variability of transcutaneous oxygen tension (tcPO2) in patients with diabetes mellitus and peripheral arterial occlusive disease who were at risk for chronic foot ulceration. METHODS: The tcPO2 was measured in the morning once daily for 3 consecutive days in 10 male patients with diabetes mellitus who were hospitalized. The mean age of the patients was 65 +/- 13 years, and they had a mean duration of diabetes mellitus of 33 +/- 6 years. The tcPO2 was measured at a reference point at the chest (I2 dx), the dorsum of the foot, and in the first intermetatarsal space. Measurements of tcPO2 in the first intermetatarsal space were performed before and during inhalation of 100% oxygen. RESULTS: The mean tcPO2 was higher (P <.001) at I2 dx (56 +/- 10 mm Hg) than at the dorsum of the foot (25 +/- 19 mm Hg) and first intermetatarsal space (27 +/- 20 mm Hg). tcPO2 increased (P <.001) during inhalation of 100% oxygen, whereas the increase was severely reduced in three patients with tcPO2 less than 10 mm Hg at baseline. A reasonably good day-to-day variability of tcPO2 was seen; the linear relations between tcPO2 investigated on days 1, 2, and 3 were highly significant (P =.0001) at each measuring site, and no systematic differences were seen between the repeated measurements (analysis of variance; P =.13 to.85). CONCLUSION: The results show an acceptable day-to-day variability of tcPO2, both at baseline and during oxygen inhalation, in patients with diabetes mellitus and peripheral arterial occlusive disease. PMID- 11496281 TI - Postoperative and amputation-free survival outcomes after femorodistal bypass grafting surgery: findings from the Department of Veterans Affairs National Surgical Quality Improvement Program. AB - PURPOSE: A noncardiac surgery risk model was used as a means of analyzing variations in postoperative mortality and amputation-free survival for older veterans undergoing femorodistal bypass grafting surgery. METHODS: A prospective cohort study was undertaken in 105 Veterans Affairs (VA) hospitals at the time of index operation from 1991 to 1995. Each patient was linked to subsequent hospitalizations, major amputation surgery, and survival through 1999. Logistic regression and proportional hazards models were used as a means of developing risk indices on the basis of risk factors from the VA National Surgical Quality Improvement Program. A total of 4288 male veterans 40 years or older underwent artificial, vein, or in situ bypass grafting surgery at the femoral to tibial level. The main outcome measures were 30-day postoperative mortality and amputation-free survival. RESULTS: Approximately half of all patients had undergone an earlier revascularization or amputation at any level for vascular disease. The 30-day postoperative mortality rate was 2.1% and varied greatly between mortality risk index quartiles (0.6%-5.2%). In a median 44.3 months of follow-up, surviving patients had 17,694 subsequent VA hospitalizations, 1147 patients (26.7%) underwent subsequent major amputation, and 1913 patients (44.6%) died. The overall survival probability was 88% at 1 year and 63% at 5 years; 1- and 5-year (any sided) limb salvage rates were 87% and 74%, respectively, for patients who underwent a femoropopliteal bypass grafting procedure, compared with 77% and 63%, respectively, for patients who underwent a tibial bypass grafting procedure. When amputation and death were combined as end points, amputation-free survival probability rates at 1, 3, and 7.5 years were 74%, 56%, and 29%, respectively. Patients with the best 20% survival risk scores had observed mean survival probability rates 30% higher than patients in the poorest 20% of survival risk. CONCLUSION: Risk indices derived from the preoperative workup may be of use to clinicians in assessing and communicating risk and prognosis. Risk adjustment of outcomes is critical for evaluating future disease management initiatives for patients with advanced peripheral arterial disease. PMID- 11496282 TI - Association of intraluminal thrombus in abdominal aortic aneurysm with local hypoxia and wall weakening. AB - PURPOSE: Our previous computer models suggested that intraluminal thrombus (ILT) within an abdominal aortic aneurysm (AAA) attenuates oxygen diffusion to the AAA wall, possibly causing localized hypoxia and contributing to wall weakening. The purpose of this work was to investigate this possibility. METHODS: In one arm of this study, patients with AAA were placed in one of two groups: (1) those with an ILT of 4-mm or greater thickness on the anterior surface or (2) those with little (< 4 mm) or no ILT at this site. During surgical resection but before aortic cross-clamping, a needle-type polarographic partial pressure of oxygen (PO2) electrode was inserted into the wall of the exposed AAA, and the PO2 was measured. The probe was advanced, and measurements were made midway through the thrombus and in the lumen. Mural and mid-ILT PO2 measurements were normalized by the intraluminal PO2 measurement to account for patient variability. In the second arm of this study, two AAA wall specimens were obtained from two different sites of the same aneurysm at the time of surgical resection: group I specimens had thick adherent ILT, and group II specimens had thinner or no adherent ILT. Nonaneurysmal tissue was also obtained from the infrarenal aorta of organ donors. Specimens were subjected to histologic, immunohistochemical, and tensile strength analyses to provide data on degree of inflammation (% area inflammatory cells), neovascularization (number of capillaries per high-power field), and tensile strength (peak attainable load). Additional specimens were subjected to Western blotting and immunohistochemistry for qualitative evaluation of expression of the cellular hypoxia marker oxygen-regulated protein. RESULTS: The PO2 measured within the AAA wall in group I (n = 4) and group II (n = 7) patients was 18% +/- 9% luminal value versus 60% +/- 6% (mean +/- SEM; P <.01). The normalized PO2 within the ILT of group I patients was 39% +/- 10% (P =.08 with respect to the group I wall value). Group I tissue specimens showed greater inflammation (P <.05) compared with both group II specimens and nonaneurysmal tissue: 2.9% +/- 0.6% area (n = 7) versus 1.7% +/- 0.3% area (n = 7) versus 0.2% +/- 0.1% area (n = 3), respectively. We found similar differences for neovascularization (number of vessels/high-power field), but only group I versus control was significantly different (P <.05): 16.9 +/- 1.6 (n = 7) vs 13.0 +/- 2.3 (n = 7) vs 8.7 +/- 2.0 (n = 3), respectively. Both Western blotting and immunohistochemistry results suggest that oxygen-regulated protein is more abundantly expressed in group I versus group II specimens. Tensile strength of group I specimens was significantly less (P <.05) than that for group II specimens: 138 +/- 19 N/cm2 (n = 7) versus 216 +/- 34 N/cm2 (n = 7), respectively. CONCLUSION: Our results suggest that localized hypoxia occurs in regions of thicker ILT in AAA. This may lead to increased, localized mural neovascularization and inflammation, as well as regional wall weakening. We conclude that ILT may play an important role in the pathology and natural history of AAA. PMID- 11496284 TI - Simple geometric characteristics fail to reliably predict abdominal aortic aneurysm wall stresses. AB - PURPOSE: The treatment of patients with abdominal aortic aneurysms (AAAs) is typically based on the potential for rupture. Current rupture assessments are in turn based on statistics from aggregate populations and are incapable of providing precise risk estimates for individual AAAs. Significant benefit could be realized if rupture potential for individual AAAs could be reliably determined on the basis of simple geometric characteristics or the results of symmetric thin shell analysis. This study seeks to determine whether it is possible to estimate wall stresses by use of these simple measures. METHODS: Linear finite element analysis was used to estimate the distribution of von Mises stresses in a series of homogeneous, isotropic, three-dimensional AAA models subject to static loading and assumed to have zero residual stresses. The magnitude of the peak stress was tabulated for each model along with the following characteristics: aneurysm volume; maximum diameter; maximum radius; maximal wall distention; aspect ratio (ratio of greatest anteroposterior diameter to transverse diameter); local radii of curvature (in both longitudinal and circumferential directions); and maximum symmetric thin-shell stress estimates (on the basis of the meridional contour). The relationship between peak stress and each of the characteristics was assessed by use of Spearman rank correlation coefficients, with values less than 0.95 interpreted as signifying unreliable associations. RESULTS: Peak stresses in the individual models ranged from 1.79 x 10(6) dyne/cm2 to 15.1 x 10(6) dyne/cm2. The circumferential and longitudinal radii of curvature were frequently able to predict the locations of high stress, but were unreliable in predicting the magnitude of peak stress. The aspect ratio showed the strongest correlation with peak wall stress (r = 0.88, 95% CI, 0.68-0.96), whereas the other characteristics showed even less correlation. Symmetric thin shell analysis accurately predicted stresses in axially symmetric models, but it was incapable of predicting either the location or magnitude of peak stress in asymmetric models. CONCLUSIONS: Simple geometric criteria and symmetric thin shell analyses are unreliable in predicting AAA stresses. Future attempts to estimate wall stress and assess risk of rupture for individual AAAs may require detailed three-dimensional modeling. PMID- 11496283 TI - Effects of anastomotic angle on vascular tissue responses at end-to-side arterial grafts. AB - OBJECTIVE: Hemodynamics has been implicated in the late failure of arterial bypass grafts, which frequently occurs at the distal anastomosis site. This study was designed to assess the relationship between local hemodynamics and pathologic responses of the distal anastomosis by manipulation of the angle of anastomosis of the graft, a major determinant of local hemodynamics. METHODS: End-to-side anastomoses of the right carotid to the left carotid arteries of rabbits were performed at anastomotic angles of less than 10 degrees (acute), 45 degrees (intermediate), or 90 degrees (right angle), and then the upstream left carotid arteries were ligated to simulate pathologic occlusion. We examined tissue responses on the wall of the recipient vessel opposite the anastomosis site (the bed), where unusual hemodynamic forces are imposed. RESULTS: Three months after surgery, intimal thickening was observed on the upstream portion of the acute, and more rarely, the intermediate anastomoses only. Medial thinning caused by loss of cells and matrix, and an aneurysm-like dilation, was observed in the right angle and some intermediate anastomoses, but not in the acute anastomoses. En face confocal microscopy at 3 weeks after surgery revealed severe disruption of the internal elastic lamina in all anastomotic models. Zymography and Western immunoblotting demonstrated gelatinolytic activity, caused by expression and activation of MMP-2, that was lowest in the acute anastomoses, higher in the intermediate anastomoses, and highest in the right-angle anastomoses. CONCLUSIONS: We infer that very different pathologic changes to the vessel wall are elicited when local hemodynamics is manipulated by altering the anastomotic branch angle. PMID- 11496285 TI - The expression and localization of membrane type-1 matrix metalloproteinase in human abdominal aortic aneurysms. AB - BACKGROUND AND OBJECTIVE: Matrix metalloproteinase-2 (MMP-2) degrades both fibrillar collagens and elastin. MMP-2 is secreted as a latent 72-kd proenzyme that must be proteolytically processed to the 62-kd active form. In our laboratory we demonstrated a significant increase of active, matrix-bound MMP-2 in abdominal aortic aneurysmal (AAA) tissue compared with nonaneurysmal aorta with arteriosclerotic occlusive disease and normal aortic tissue. This increase in active MMP-2 is considered to be important in aneurysm pathogenesis, but the mechanism of its activation in aortic tissue is unknown. Membrane type-1 MMP (MT 1 MMP) is known to be an activator of MMP-2. The purpose of this study was to determine MT-1 MMP expression and its involvement in pro-MMP-2 activation in human aneurysmal tissue. METHODS: Infrarenal aortic tissue was obtained during the surgical repair of AAAs or the bypass of aortoiliac occlusive disease, or from nondiseased aorta, and the expression of MT-1 MMP messenger RNA was determined with Northern blot analysis. MT-1 MMP protein was determined with immunoblot and immunohistochemistry. The ability of aortic tissue to activate pro MMP-2 was analyzed by incubating aortic tissue with exogenous radiolabeled pro MMP-2. RESULTS: MT-1 MMP messenger RNA and protein are increased in AAA (P <.05) compared with arteriosclerotic occlusive disease and normal aortic tissue. Immunohistochemical analysis localized MT-1 MMP to aortic smooth muscle cells and macrophages in aneurysmal tissue. AAA tissue demonstrated a greater capacity to activate exogenous pro-MMP-2 compared with atherosclerotic and normal aortic tissue (P <.05). CONCLUSION: These studies demonstrate that MT-1 MMP is increased in AAA tissue and suggest that it may be important in AAA pathogenesis through its ability to activate pro-MMP-2 PMID- 11496286 TI - Acute arterial thrombosis causes endothelial dysfunction: a new paradigm for thrombolytic therapy. AB - PURPOSE: The goals of this study were to delineate the time course of endothelial dysfunction after arterial thrombosis, to determine the cause of endothelial dysfunction in this setting, and to determine whether modulating standard thrombolytic therapy would ameliorate the thrombosis-mediated endothelial dysfunction. METHODS: Male adult rats underwent infrarenal aortic occlusion by means of clip ligature to induce arterial thrombosis. After 30 minutes, 1, 2, and 3 hours, ring segments from the infrarenal aorta were harvested and placed into physiologic buffer baths. With the use of a force transducer, both endothelial dependent relaxation (EDR) and endothelial-independent relaxation (EIR) were measured. Endothelial function and presence were determined by means of factor VIII immunohistochemical staining. Endothelial morphology was evaluated with scanning electron microscopy (SEM). Nitric oxide (NO) levels were determined with a chemiluminescent assay of its nitrite/nitrate metabolites (NO(x)). Standard thrombolytic therapy with urokinase (UK) was infused into thrombosed aortic ring segments and compared with UK supplemented with both low-dose L -arginine (2 mmol) and high-dose L -arginine (20 mmol). RESULTS: Arterial thrombosis decreases EDR. The nadir of EDR occurs 1 hour after thrombosis (mean +/- SE, 13% +/- 6.4% vs 94% +/- 2.6% for controls, P <.005), with persistent lowering of EDR as long as 3 hours after thrombosis. EIR is preserved, and vasoconstriction with norepinephrine or potassium buffer is unaltered. Both endothelial function and presence (n = 6 per group) were documented by means of factor VIII immunohistochemistry. An intact monolayer of endothelium at all time intervals after thrombosis was revealed by means of SEM analysis. No differences between control and thrombosed specimens were revealed by means of the grading of SEM images. Local NO(x) levels were lower after 1 hour of thrombosis, with an increase higher than baseline values at 3 hours. The addition of low-dose L arginine resulted in a minor increase in EDR. However, high-dose L -arginine resulted in a significant increase in EDR versus controls receiving UK alone (64% +/- 6.3% vs 38% +/- 4.4%, P <.05). Correspondingly, local NO(x) levels were 20 fold higher after the high-dose L -arginine supplementation when compared with UK thrombolysis alone (2.8 +/- 0.52 micromol/L vs 0.133 +/- 0.02 micromol/L, n = 6 samples/group, P <.005). CONCLUSION: Acute arterial thrombosis causes endothelial dysfunction, without causing endothelial cell loss. Endothelial function reaches a nadir after 1 hour of thrombosis. EIR and vasoconstriction remain unaffected, indicating normal smooth muscle cell function. NO(x) levels suggest that NO levels are decreased acutely after thrombosis. Supplementing standard thrombolytic therapy with the NO precursor, l-arginine, ameliorates the endothelial dysfunction seen after acute thrombosis by increasing local NO production. PMID- 11496287 TI - Identification of the critical spinal arteries with F wave-polysynaptic response complex monitoring: an experimental study. AB - PURPOSE: The main cause of paraplegia after surgery for descending thoracic aortic aneurysms and thoracoabdominal aortic aneurysms is spinal cord ischemia caused by ligation of the critical spinal arteries that are associated with the great radicular artery (GRA). In this experimental study, we attempted to identify the critical spinal arteries with F wave-polysynaptic response complex (FPC) monitoring. The FPC, which we are using as a monitor of spinal cord ischemia, is myogenic potentials evoked by the tibial nerve stimulation through the anterior horn cells. METHOD: In 18 rabbits, infrarenal lumbar arteries were clamped until there was an FPC amplitude reduction (FPCAR) of at least 60%. The rabbits were classified according to the number of arteries clamped to produce the FPCAR (group A, 1 artery; group B, 2 arteries; group C, 3 arteries). Selective angiography of each lumbar artery was performed in all rabbits. RESULTS: An FPCAR was observed in all 18 rabbits. In all nine rabbits in group A, the FPCAR was caused by the clamping of one particular lumbar artery. The GRA was shown by means of selective angiography to originate directly from this lumbar artery. In the nine rabbits in groups B and C, 11 FPCARs were observed. Of these, nine FPCARs in nine rabbits were caused by the clamping of lumbar arteries that included the vessel from which the GRA originated. CONCLUSION: Temporary clamping of lumbar arteries with FPC monitoring can be used as a means of detecting the critical spinal arteries from which the GRA originates. PMID- 11496288 TI - Dual cell seeding and the use of zymogen tissue plasminogen activator to improve cell retention on polytetrafluoroethylene grafts. AB - OBJECTIVE: The purpose of this study was to enhance the retention of seeded endothelial cells (EC) on prosthetic vascular grafts. Dual-layer EC and smooth muscle cell (SMC) seeding and gene transfer of a zymogen tissue plasminogen activator gene (tPA) into seeded EC were studied. METHODS: Polytetrafluoroethylene (PTFE) grafts were precoated with fibronectin, seeded with SMC followed by EC a day later, and then, 24 hours later, exposed to an in vitro flow system for 1 hour. Cell retention rates were determined for grafts seeded with EC only, a dual layer of EC on top of SMC, EC transduced with wild type tPA, and EC transduced with zymogen tPA. RESULTS: Seeding efficiency of PTFE pretreated with fibronectin was 260 +/- 8 cell/mm(2). After exposure to flow, only 39% +/- 14% of the EC were retained when EC were seeded alone, whereas 73% +/- 22% of EC remained on grafts when EC were seeded on top of SMC (P <.001, n = 10). The enzyme activity of a mutant zymogen tPA in absence of fibrin was 14 +/- 1 IU/mL, which is 3.6-fold lower than that in the presence of fibrin (50 +/- 19 IU/mL), whereas fibrin has no effect on the wild-type tPA activity. EC expressing a high level of wild-type tPA had a lower retention rate (37%) when compared with normal EC (45%). EC expressing the mutant zymogen tPA had an improved retention rate (54%, P =.001, n = 10) in absence of fibrin, whereas its retention rate was reduced to 43% when the cells were exposed to fibrin. CONCLUSION: SMC seeded between EC and PTFE improves EC retention in vitro. Transduction of zymogen tPA increases thrombolytic ability of seeded cells with less adverse impact on cell retention than wild-type tPA. PMID- 11496289 TI - Overexpression of the focal adhesion kinase (p125FAK) in the vascular smooth muscle cells of intimal hyperplasia. AB - PURPOSE: The migration and proliferation of vascular smooth muscle cells (VSMCs) are important events in the development of intimal hyperplasia (IH). The focal adhesion kinase (FAK) gene encodes a protein tyrosine kinase (p125FAK) involved in signal transduction pathways used in cell adhesion, motility, and proliferation. Because alterations in these cellular processes are thought to occur in VSMCs during IH, we studied FAK expression in healthy arteries and veins in comparison with that in pathologic vessels containing IH. METHODS: To determine p125FAK expression at the cellular level, we developed a monoclonal antibody that specifically detected FAK in formalin-fixed, paraffin-embedded tissue sections (5 microm) and analyzed the levels of FAK expression in human arteries and veins. Specificity of monoclonal antibody 4.47 was demonstrated by means of immunofluorescence microscopy showing FAK-specific staining at focal adhesions of healthy human vascular smooth muscle cells (AoSMCs). By using immunohistochemistry techniques, we analyzed the expression of p125FAK in 25 adult human vascular tissue samples from individual patients, which contained a histologically confirmed healthy artery, vein, or IH. RESULTS: FAK expression in healthy and pathologic human vascular tissue was localized predominantly within VSMC cytoplasm. In healthy human artery and vein, borderline FAK expression was detected in the media of seven of 17 vessels and undetectable in the remainder of specimens. However, in vessels containing IH, FAK was overexpressed in the pathologic VSMC populations at moderate-to-strong levels in eight of eight specimens. The levels of FAK expression were directly correlated with structures containing IH, and the results of FAK staining intensity and the percentage of positive cells in these samples were significantly increased compared with normal vascular tissue levels (P <.05, Student t test). CONCLUSION: These results provide the first evidence that FAK is overexpressed in VSMCs involved in IH and suggest that FAK upregulation may be part of a mechanism for migration and proliferation of VSMCs during this process. Furthermore, the dramatic upregulation of FAK in IH and the relative lack of expression in healthy vessels suggest that FAK may be a rational target for controlling IH. PMID- 11496290 TI - Aortic Slimgraft: ex vivo and in vivo study. AB - In this article we describe a method for assembling an endovascular graft within the aorta with a graft and then a stent introduced sequentially over a guidewire as separate components. In the ex vivo study, an endovascular graft that was 25 mm in diameter was introduced through a 9F introducer. In the in vivo study, smaller endovascular grafts were placed in the aortas of four healthy pigs and five pigs with aortic aneurysms. Our data suggest that this very low profile system may have significant clinical implications because it converts aortic repair into a percutaneous procedure. PMID- 11496291 TI - Detection of isolated hook fractures 36 months after implantation of the Ancure endograft: a cautionary note. AB - Two cases of delayed (36-month) Ancure hook fracture are reported in patients who experienced a decrease in aneurysm size and no evidence of endoleak. Both devices used redesigned hooks and are otherwise identical to those devices currently used in clinical practice. Notably, hook fractures were not visualized on all abdominal radiographic views, nor were they noted on the final "institutional" report by the reviewing radiologist. Careful clinical follow-up with multiple view abdominal radiographs remains essential for all patients treated with an endovascular graft, with particular attention directed to the integrity of the metal components. The broader clinical significance of this observation with respect to the Ancure endograft remains to be defined. PMID- 11496292 TI - An operative method for surgical revision of a late failure after endovascular repair of an abdominal aortic aneurysm. AB - Endoleak is a well-recognized complication of endovascular aortic exclusion stent grafts. The Vanguard device is anchored into the proximal aorta by an expandable stent surmounted by a circular array of metal hooks. Attempted removal of the whole prosthesis may therefore be technically difficult and result in damage of the aortic wall. Surgery for endoleak may be complicated and carry a mortality rate of 20% to 40%. We describe a simple surgical technique for repair of endoleaks associated with a Vanguard stent-graft, which would be applicable to other such devices. This method involves minimal disruption of the proximal stent and avoids problems that may arise from an attempt to completely remove the endovascular device. PMID- 11496293 TI - Superior vena cava syndrome: relief with a modified saphenojugular bypass graft. AB - Superior vena cava (SVC) syndrome is a disabling and potentially life-threatening complication. SVC revascularization can be achieved by means of endovascular or direct surgical reconstructions. In the patient on whom we report, these two options were not possible, and a peripheral venous bypass grafting procedure was done with a technical innovation. Right upper-extremity swelling developed in a 55-year-old woman after radiation therapy for lung carcinoma. A left subclavian vein Port-A-Cath induced extensive thrombosis of the left innominate, axillosubclavian, and jugular veins. She was referred to our institution with very symptomatic SVC syndrome after two failed endovascular interventions. The occlusion of both innominate veins and chronic thrombus extending into the left axillosubclavian and internal jugular veins was confirmed by means of a venogram. A third endovascular attempt failed. The presternal skin had severe radiation induced damage precluding direct SVC reconstruction. A bypass grafting procedure from the right internal jugular to the femoral vein was performed with spliced bilateral greater saphenous veins tunneled inside an externally supported expanded polytetrafluoroethylene graft. Postoperatively, the patient had no symptoms, and graft patency was confirmed by means of duplex ultrasound scanning. A saphenojugular bypass grafting procedure can offer prompt and durable relief of SVC syndrome when endovascular or direct surgical reconstructions are not possible. This rarely used peripheral venous bypass grafting procedure was modified by tunneling the vein graft inside an externally supported polytetrafluoroethylene graft to prevent kinking or compression. PMID- 11496294 TI - Spontaneous aortic dissection and rupture in a patient with neurofibromatosis. AB - Vascular involvement in neurofibromatosis type I is a well-recognized but rare feature of the disease. This case report is the first such describing acute aortic dissection and rupture in a patient who also had incidental aneurysms of his carotid and iliac arteries. A review of the literature about pathogenesis and management strategies is also presented. PMID- 11496295 TI - A technique for adequate coverage of the proximal suture line during abdominal aortic aneurysm repair. AB - The proximal suture line is a vulnerable area after abdominal aortic aneurysm repairs. This area has been implicated in various postoperative complications, such as pseudoaneurysm formation, graft-enteric fistula, and suture line disruption. We present a technique that provides safe and adequate coverage of this suture line by using the aneurysm sac. This technique is derived from the z plasty technique used for scar revision. The technique is illustrated with detailed line drawings. None of the patients in whom we used this technique have had any complications related to the proximal suture line. PMID- 11496297 TI - Outcome assessment: a new approach to the maintenance of board certification in vascular surgery. PMID- 11496298 TI - Regarding "does surgical correction of the superficial femoral vein valve change the course of varicose disease?". PMID- 11496300 TI - Expression of DNA mismatch repair proteins hMSH2 and hMLH1 and the cyclin G1 inhibitor, p21(waf1/cip1) in pediatric tumors: correlation with response to therapy. AB - The expression of the DNA mismatch repair proteins hMSH2 and hMLH1 and p21(waf1) the cyclin G1 inhibitor, may determine response of adult cancers to anti-cancer drugs, that include alkylating agents and platinum-based drugs. The role of DNA mismatch repair proteins (hMSH2 and hMLH1) and p21(waf1) in pediatric tumor responses to chemotherapy and irradiation is described in the present study of 23 pediatric solid cancers (4 wilms' tumors, 9 neuroblastomas, 3 hepatoblastomas, 3 lymphomas and 4 sarcomas) using immunohistochemical methods. Immunostaining was scored for intensity (0-3) and extent (0-3). Most tumors stained strongly for hMSH2 and weakly or negative for hMLH1. All the hMLH1 negative tumors (1 wilms', 1 hepatoblastoma, 1 sarcoma, 2 lymphomas and 2 neuroblastomas) achieved complete response. p21(waf1) negative and positive tumors achieved relatively similar treatment response. The results suggest that the expression of DNA mismatch repair proteins hMLH1 and hMSH2, and p21(waf1) do not influence individual cancer responses to treatment and the results may reflect the use of multiple drugs and irradiation that cause many different types of DNA damage. PMID- 11496299 TI - Adenoviral-mediated gene therapy of human bladder cancer with antisense interleukin-8. AB - We previously demonstrated the importance of interleukin-8 (IL-8) as a mediator of angiogenesis, tumorigenicity, and metastasis of transitional cell carcinoma (TCC) of the bladder. In the present study, we evaluated the feasibility of adenoviral mediated antisense IL-8 gene transfer (Ad IL-8-AS) as therapy for established TCC. In vitro, Ad IL-8-AS inhibited endothelial cell proliferation and enhanced endothelial cell apoptosis. The highly metastatic human TCC cell line 253J B-V(R) was implanted into the subcutis of athymic nude mice, and intralesional therapy with Ad IL-8-AS commenced when the tumors reached a diameter between 5 and 7 mm. Tumor growth was significantly inhibited compared with therapy in controls (saline and beta-galactosidase adenovirus). Ad IL-8-AS therapy decreased the in vivo expression of IL-8 and matrix metalloproteinase type 9 (MMP-9), reduced microvessel density, and enhanced endothelial cell apoptosis. These results indicate that Ad IL-8-AS therapy targets both tumor cells and host endothelial cells resulting in endothelial cell apoptosis and significant inhibition of tumor growth. PMID- 11496301 TI - Combined evaluation of NGF and p75NGFR expression is a biomarker for predicting prognosis in human invasive ductal breast carcinoma. AB - Nerve growth factor (NGF) is a member of the neurotrophin family and is essential for the differentiation and maintenance of neural cells. Recently, it has been reported that NGF is involved in the growth of breast cancer. On the other hand, two types of NGF receptors have been identified, a low-affinity receptor, p75NGFR, and a high-affinity receptor, TrkA. NGF-p75NGFR interaction is known to play an important role in apoptosis, whereas NGF-TrkA interaction is responsible for the survival of neural cells. We examined the relationship between clinicopathological factors, Ki-67 index, apoptotic index and the immunohistochemical expression of NGF, TrkA and p75NGFR in 71 invasive ductal breast carcinoma (IDBC) specimens. Our data indicate that positive Ki-67 expression (a labeling index exceeding 30%) correlates significantly with the positive expression of NGF (p=0.0091). Moreover, the apoptotic index was found to correlate with a strong expression of p75NGFR. Furthermore, patients who were NGF positive and p75NGFR negative had significantly poorer disease-free survival rates (p=0.0165). In contrast, those who were NGF negative and p75NGFR positive had significantly more favorable outcomes (p=0.0191). These findings suggest that a combined evaluation of NGF and p75NGFR expression is a predictive factor in the prognosis of IDBC patients. PMID- 11496302 TI - Carbohydrate antigens as a risk factor for hematogenous recurrence of esophageal squamous cell carcinoma patients. AB - The patients with hematogenous recurrence have a poor prognosis in esophageal squamous cell carcinoma (SCC). Recent developments have suggested the important role of sialyl Lewis A and X in the step of hematogenous metastasis. To evaluate the role of two carbohydrate antigens on hematogenous recurrence in esophageal SCC, we examined clinicopathological features and the expression of sialyl Lewis A and X in 125 patients retrospectively. Thirty-three out of 125 patients had hematogenous recurrence after curative esophagectomy. sLeA and sLex expression correlated with hematogenous recurrence (p=0.026, p=0.043 respectively), and sLeA expression was correlated with pM (lymph). Cox proportional hazards model and logistic regression analysis revealed that pN and pM lymph were significant prognostic factors and predictive factors of hematogenous recurrence in esophageal SCC. Although, neither sLeA nor sLeX were significant factors affecting survival in esophageal SCC, logistic regression analysis demonstrated that increased tumoral expression of sLeA and sLeX were risk factors of hematogenous recurrence in esophageal SCC. In conclusion, patients with lymph node metastasis, in particular those with distant lymph node metastasis, the expression of sLeA and sLeX may suggest a higher incidence of hematogenous recurrence in esophageal SCC. New strategies for specific inhibition of cancer cell attachment to vascular endothelial cells should be considered for the patients with the increased tumoral expression of either sLeA or sLeX. PMID- 11496303 TI - The influence of hormones on CD44 expression in endometrial and breast carcinomas. AB - The expression of distinct variant isoforms of the cell surface glycoprotein CD44 (CD44v) has been found to be associated with metastatic potential of rodent adenocarcinoma cells and with an altered prognosis in several types of human cancer. In hormone-dependent gynecological cancers, different CD44v expression patterns have been observed. The influence of ovarian steroid hormones and their antagonists on CD44v expression is still unclear, since there are only retrospective correlation studies so far. Therefore, we examined the CD44 mRNA expression in a standardized stimulation experiment in a number of breast and endometrial carcinoma cell lines varying in estrogen receptor (ER) status. Higher CD44 overall expression was observed in ER positive endometrial and breast carcinoma cell lines when compared to corresponding ER negative cell lines. The number and composition of alternatively spliced isoforms showed no clear correlation to the ER expression status. Three CD44v isoforms were detected in all cell lines expressing CD44v, two of which have not been reported previously in normal endometrial cells. These isoforms may have specific functions in this type of carcinoma. In the second part of the study, the influence of (anti-) hormones on CD44 expression in endometrial carcinoma cell lines was examined. CD44 overall expression showed an increase when the cells were grown in medium containing fetal calf serum (FCS) as compared to cells maintained in medium-free of FCS. CD44 expression was transiently increased by estradiol (1 h). The CD44 splice pattern of endometrial cancer cell lines RL95-2 and Hec-1-A, after treatment with (anti-) hormones showed constant and high expression rates for distinct CD44v-isoforms such as CD44E (CD44v8-v10). Only certain weakly expressed isoforms changed their expression level during the experimental period, but no direct correlation to hormone treatment was observed. In conclusion, estradiol or FCS increase CD44 overall expression, but there seems to be no direct influence of ovarian steroid hormones on the CD44v splice machinery in endometrial carcinoma cell lines. PMID- 11496304 TI - Detection of Epstein-Barr virus in gastrectomy specimens. AB - Epstein Barr virus (EBV) has been reported to be present in a minority of gastric carcinomas and may be implicated in its pathogenesis. This study was aimed at determining the occurrence of EBV in 43 consecutive gastrectomy specimens with a variety of benign and malignant lesions. In situ hybridisation was used for detection of EBER RNA, the marker for latent EBV infection. Only 1/20 (5%) gastric cancers was EBER positive; a moderately differentiated adenocarcinoma with a heavy lymphocytic infiltration. The interstitial lymphocytic infiltrate was predominantly of B cell type, but the majority of lymphocytes overlying the tumour cells were CD8+ T cells. The other gastric lesions examined, which included 15 peptic ulcers, 6 stromal tumours and 2 lymphomas, were all EBER negative. Using a biotin detection system, scattered EBER positive cells were seen in adjacent normal gastric and/or duodenal mucosa in 9 sections from 8 cases (i.e., in 19% of all 43 cases examined). However, on using a digoxygenin detection system, no reactivity was found in these normal cells. An immunoperoxidase stain for chromogranin A showed that these apparently 'EBER positive' cells corresponded to normal chromogranin positive neuroendocrine cells within the gastric and duodenal mucosa. We conclude that EBV infection occurred only in the lymphoepithelioma type of gastric carcinoma and was absent from the other adenocarcinomas and from normal and benign tissues. The occasional EBER positive reaction encountered in normal cells was probably the result of a false signal arising from neuroendocrine cells as a consequence of the biotin containing detection system. PMID- 11496305 TI - Genomic instability and breast cancer. AB - The loss of genomic stability is accepted as being one of the most important aspects of cancer. The correlation between genomic instability and cancer proneness in cases of known genetic syndromes (e.g. ataxia telengectasia, Fanconi anemia) is well established. This study was conducted to assess genomic instability in 19 patients with sporadic breast cancer. We used the comet assay on the lymphocytes of patients before radiotherapy and/or chemotherapy. The alkaline comet assay (single-cell gel electrophoresis) is a sensitive and rapid method for detecting DNA damage (single strand breaks and alkali-labile sites) in G(0) cells, at a single-cell level. This assay was achieved in vitro without irradiation and after exposure (dose ranging from 50 cGy to 5 Gy). The results show that the patients have higher baseline values than controls. At 2 Gy, the mean tail moment, score and the percentage of DNA in the tail increase for both groups but these values are much higher for patients. Our results show that the lymphocyte DNA of cancer patients is more damaged than that of controls with or without irradiation. Our hypothesis is that this baseline DNA damage reflect a genomic instability in sporadic breast cancer. This instability seems to increase after in vitro irradiation. PMID- 11496306 TI - Radiosensitization in sarcoma cell lines with a p53 missense mutation correlates with prevention of irradiation G2/M arrest but not with induction of apoptosis. AB - We analysed the effects of caffeine and taxol on the radiobiological behaviour of two human sarcoma cell lines (RD, SK-LMS-1) each with a p53 missense mutation. Treatment with 2 mM caffeine resulted in an inhibition of the irradiation induced G2/M arrest in both cell lines. This effect was coupled with a radiosensitization in cell line SK-LMS-1 after an irradiation with 6 Gy (enhancement factor of 5.0). However, the effect of radiosensitization was not correlated with an induction of apoptosis. Incubation with 20 nM taxol increased the irradiation induced apoptosis almost 3-fold in cell line SK-LMS-1, but not in cell line RD. However, taxol had no effect on the irradiation induced G2/M arrest or radiosensitivity in either cell line. The results support the hypothesis that the prevention of irradiation induced G2/M arrest but not the induction of apoptosis plays a critical role in determining radiosensitivity in sarcoma cell lines with p53 mutations. PMID- 11496307 TI - Pharmaco-economic study of intensive chemotherapy with peripheral blood progenitor cell support for advanced breast cancer. AB - Patients with high-risk breast cancer may benefit from dose-escalated chemotherapy. The rationale for high-dose chemotherapy with stem-cell rescue (HDC SCR) in breast cancer is based on the principles of dose response and dose intensity. However, several results of properly randomized and prospective studies are necessary to determine the interest of HDC-SCR. The objective of this study, realised in the Anticancer Center of Montpellier, was to evaluate the cost of this type of transplantation. In this retrospective study, we analysed 30 patients treated for an advanced breast cancer between October 1995 and June 1998. We collected the data from the induction chemotherapy cycle (followed by cytapheresis) to the end of the hospitalisation for autograft. The mean total cost was US $25,845 per patient: US $6453 for drugs, US $4720 for transfusions, US $1865 for laboratory services and blood tests, US $5585 for staff pay, US $774 for material, US $1211 for administration cost, US $1111 for logistic cost, US $998 for structure cost and US$1578 for other. Antibiotics, granulocyte-colony stimulating factor, chemotherapy, transfusions and nutrition represent respectively 18% (US $1962), 14% (US $1590), 13% (US $1437), 42% (US $4720), 11% (US $1191) of the medication and blood products cost. The results of this study may help in identifying targets for cost reduction. PMID- 11496308 TI - Bcl-2 expression in breast cancer is down-regulated by trans-arterial administration of chemotherapeutic agents. AB - Bcl-2 is one of the cytoplasmic oncoproteins, and has been shown to suppress apoptotic cell death. In this study, we investigated the relationship between Bcl 2 expression and effects of chemotherapeutic agents on human breast cancer cells. We examined 26 surgically resected breast tumors with preoperative trans-arterial administration of chemotherapeutic agents and 30 control cases using immunohistochemical methods. In all 26 cases in the chemotherapy group, the breast cancer cells were focally degenerated to various degrees, associated with inflammation and stromal desmoplastic changes. Bcl-2 expression was found in 46% (12/26) of the chemotherapy group and in 67% (20/30) of controls. Of the 12 Bcl-2 positive cases in the chemotherapy group, 5 were diffusely positive [Bcl-2(2+)] and 7 were focally positive [Bcl-2(+)]. Of the 20 Bcl-2-positive cases in the control group, 18 were diffusely positive and 2 were focally positive. We speculate that Bcl-2 expression was down-regulated by trans-arterial administration of chemotherapeutic agents and was associated with apoptosis and degeneration of breast cancer cells. PMID- 11496309 TI - Cyclooxygenase-2 expression in an adenoid cystic carcinoma of the uterine cervix. AB - Adenoid cystic carcinoma (ACC) of the uterine cervix is extremely rare, frequently metastasizes to distant organs, and its prognosis is poorer than squamous cell carcinoma of the uterine cervix. The reasons for its poor prognosis are unclear. This case had both an ACC and a carcinoma in situ (CIS) of the uterine cervix, so the expressions of CD34, vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) were investigated in both tumors. Hysterectomy was performed on a 76-year old woman and the uterine cervical tissues immunohistochemically analyzed. Expressions of CD34 were positive in the ACC lesions but negative in the CIS ones and angiogenesis was confirmed in ACCs. Furthermore, expressions of VEGF and COX-2 were shown in ACC, but were absent in CIS. In conclusion, the expression of COX-2 in ACC may induce the expression of VEGF, increase angiogenesis and enhance tumor growth and invasion. PMID- 11496310 TI - Sudden termination of G-CSF injection leads to apoptosis of a large proportion of increased granulocytes. AB - The effectiveness of granulocyte colony-stimulating factor (G-CSF) for granulocytopenia has been widely recognized. However, although granulocyte counts rapidly increase after a few injection of G-CSF, a large proportion of the increased granulocytes disappear from peripheral blood within a few days following G-CSF withdrawal. Where do they go? In this report, the answer can be seen at a glance. Using MitoCapture and a CCD camera-equipped fluorescence microscope, we succeeded in demonstrating that G-CSF withdrawal induced loss of mitochondrial membrane potential, i.e., an early stage of apoptosis, in human peripheral granulocytes increased by G-CSF. PMID- 11496311 TI - Conservative management of placental polyp with oral administration of methotrexate. AB - Surgical treatment is usually selected for placental polyp accompanied by massive bleeding but patients wishing to conserve their fecundity require conservative management. A 35-year old nullipara was diagnosed as having placental polyp on the basis of typical episodes, and detection of placental polypeptide hormones and blood flow by Doppler ultrasonography and dynamic magnetic resonance imaging. Oral administration of methotrexate (2.5 mg, three times a day for 5 days) was repeated for three cycles because surgical treatment was rejected. Serum human placental lactogen, blood flow and the polyp disappeared sequentially following chemotherapy. This report thus advocates considering conservative management for placental polyp. PMID- 11496312 TI - Stilbene estrogen produces higher levels of mitochondrial DNA adducts than nuclear DNA adducts in the target organ of cancer (liver) of male Sprague Dawley rats. AB - We have previously demonstrated that diethylstilbestrol is metabolized to diethylstilbestrol reactive metabolites by mitochondrial enzymes in vitro. In vitro, these reactive intermediates bind to mitochondrial DNA. Here we have investigated the in vivo formation of diethylstilbestrol adducts with mitochondrial DNA and the nature of mitochondrial DNA-diethylstilbestrol adducts. Diethylstilbestrol exposure to male rats produced several adducts in mitochondrial DNA of both kidney and liver. The total relative adduct levels were 7-fold higher in mitochondrial DNA than in nuclear DNA in the target organ of cancer (liver) of Sprague Dawley rats. The chromatographic mobility of mitochondrial DNA adducts formed in vivo were similar to that of dGMP-DES quinone adducts formed in vitro. These findings suggest that mitochondrial DNA appears more susceptible to formation of diethylstilbestrol adducts than nuclear DNA, and the results suggest that obstruction of replication and/or transcription of the mitochondrial genes by covalent modifications of the mitochondrial DNA by diethylstilbestrol may produce mitochondrial genomic instability in vivo and may provide an explanation for the carcinogenic effects of DES. PMID- 11496313 TI - Molecular analysis of CDKN2 (p16) in gliomas associated with clinical data. AB - We analyzed alterations in CDKN2 in gliomas from an ethically mixed population and correlated the results with patients clinical data. We screened for methylation at CDKN2 and for microsatellite instability (MSI) and loss of heterozygosity (LOH) in the region 9p21-22 using 4 markers. We found: 3/30 (10%) cases with CDKN2-methylated gliomas; an average of 4% of MSI; and 24.5% of LOH in the region 9p21-22. Methylation of CDKN2 was only detected in patients showing high-grade gliomas with short survival. MSI and LOH in the region 9p21-22 were detected in patients showing high-grade gliomas with short survival and in one patient with a recurrent low-grade astrocytoma grade II who died from the disease after 3 years, indicating that such alterations represent poor prognosis. PMID- 11496314 TI - Transplacental effect of a 15% olive-oil diet on functional activity of immune components in the spleen and colon tumors of rat offspring. AB - We studied whether feeding pregnant female rats a 15% olive-oil diet affects the activity of lymph cells in the spleen and tumors in offspring with chemically induced colon tumors. Rat mothers were fed either a 7% corn-oil or a 15% olive oil diet. Five-week-old male offspring were divided into 3 groups. A control group was fed the 7% corn-oil diet similar to their mothers. The experimental group I was fed the 7% corn-oil diet whereas their mothers were fed the 15% olive oil diet. The experimental group II was fed the same 15% olive-oil diet as their mothers. Experimental rats were injected weekly for 8 weeks with the carcinogen, 1,2-dimethylhydrazine (DMH), 20 mg/kg b.w. Results of experiments were studied 6 months later. The area of zones in the spleen responsible for producing B and T lymphocytes were measured and the number of cells counted. The activity of lymphoid elements of the spleen and of tumors were studied using immunohistochemical methods for evaluating the synthesis of CD8(+) lymphocytes and proliferative activity of lymphocytes in spleens and tumors. Feeding pregnant or lactating mothers with the 15% olive-oil diet had no marked tumor-protective effect on chemically-induced colon cancer in offspring. Diet-dependent changes were found at the cellular level. In the spleen of control offspring, the presence of a tumor was accompanied by an increase in the number of Ki-67(+) cells and CD8(+) lymphocytes in the red pulp. In experimental group I, DMH significantly increased the total cell number and the number of CD8(+) lymphocytes in the red pulp of the spleen in both tumor-bearing and tumor-free rats. In experimental group II, the total number of lymph cells and the number of CD8(+) lymphocytes increased compared to offspring fed a control diet. Tumor formation activated the proliferative activity of lymph elements. The total number of cells in infiltrates of the colon mucosa decreased in tumor-bearing rats compared to tumor-free counterparts, and this was seen in all three dietary groups of rats. In tumors from offspring of experimental group II, only the number of CD8(+) lymphocytes increased compared to those in offspring of experimental group I. The findings indicate that feeding mothers the 15% olive oil diet had a cancer-inhibiting role in offspring, predominantly changes at the cellular level. PMID- 11496315 TI - Relationship between existence of lymphatic invasion in peritumoral breast tissue and presence of axillary lymph node metastasis in invasive ductal carcinoma of the breast. AB - Specimens obtained from 92 patients with invasive ductal carcinoma of the breast by quadrantectomy and axillary lymph node dissection were examined to evaluate the relationship between existence of lymphatic invasion in peritumoral breast tissue and presence of axillary lymph node metastasis. The number of lymphatic invasions was classified into 4 groups (ly0-3) by counting the number of peritumoral lymphatic invasions. In addition, immunohistochemistry for cytokeratin was performed to locate micrometastasis in the dissected lymph nodes. Thirty-seven (40.2%) of 92 cases had foci of lymphatic invasion and 29 (31.5%) cases revealed lymph node metastasis on initial routine examination. The rate of diagnosis of lymph node metastasis assessed by the existence of lymphatic invasion had an accuracy of 84.8%, a sensitivity of 89.7% and a specificity of 82.5%. On the other hand, all 3 cases (4.8%) with micrometastasis detected by immunohistochemistry for cytokeratin, showed lymphatic invasion. The rate of diagnosis after detection of micrometastasis increased and exhibited 88.0% accuracy. In addition, the rate of prediction of lymph node metastasis in cases with tumor larger than 15 mm was also high, and its accuracy was 88.2%. These results suggest that the assessment of peritumoral lymphatic invasion is very useful for predicting the presence of axillary lymph node metastasis including micrometastasis. They also suggest that excision specimens should be examined for lymphatic invasion, and that the results of the examination might be necessary to pick up false-negative cases and those at high risk for lymph node metastasis among patients who have not undergone lymph node dissection based on the result of sentinel lymph node biopsy. PMID- 11496316 TI - Human colon cancer produces a factor which induces the proliferation of venous endothelial cells. AB - This study was performed to investigate whether colorectal cancers produce proliferative factors for venous endothelial cells, and whether the proliferative activity is related to basic FGF and VEGF and also to the clinicopathological findings. Surgically resected specimens of 17 colorectal cancer patients were fragmented and cultured, and the supernatant was collected. A human umbilical endothelial cell line (EA-hy 926 cells) was incubated with the supernatant. The proliferative activity was examined and the levels of basic FGF and VEGF were measured. The activities were found to be significantly related to VEGF, the depth of tumor invasion and the tumor stage. PMID- 11496317 TI - Cdc6 expression as a marker of proliferative activity in brain tumors. AB - Antibodies against human proteins that regulate DNA replication such as Cdc6 and Mcm5 became available as a new marker of proliferation. We performed immunohistochemical analysis with MIB-1 and antibody against Cdc6 on 35 brain tumors, including tumors of neuroepithelial tissue, vestibular schwannomas, meningiomas, and pituitary adenomas. Median reactivity for MIB-1 was 8.8%, and that for Cdc6 was 55%. Reactivity in most brain tumors was significantly higher for Cdc6 than for MIB-1, but reactivity of Cdc6 was independent of tumor grade. Detection of Cdc6 expression might be useful for the estimation of proliferative activity in brain tumors. PMID- 11496318 TI - A randomized study comparing oral and standard regimens for metastatic breast cancer. AB - We conducted a randomized controlled trial comparing oral regimen [doxifluridine, an intermediate metabolite of capecitabine, + medroxyprogesterone acetate (MPA) + cyclophosphamide (CPA)] (Method A) with a standard regimen (5-fluorouracil + adriamycin + CPA) plus MPA (Method B) as first line chemotherapy for metastatic breast cancer. Overall response rate was 55.8% for Method A, 46.3% for Method B. The total ratio of responder and long stable disease was significantly higher with Method A (p=0.006). Median time to progression and survival were not differences between Methods. Incidence of toxicity was 56.3% with Method A and 80.0% with Method B (p=0.014). Oral regimen is more useful than standard therapy. PMID- 11496319 TI - The tumor-preventing effect of a mixture of several lactic acid bacteria on 1,2 dimethylhydrazine-induced colon carcinogenesis in mice. AB - The anti-tumor effect of a dietary supplement obtained from mixed cultures of several lactic acid bacteria was examined in the colon of tumor-inducing ICR male mice by use of a carcinogen, 1,2-dimethylhydrazine (DMH, 20 mg/kg body weight, 1 intra-muscular injection per week for 10 weeks). The animals were sacrificed either 15 weeks or 24-26 weeks after the first carcinogen injection. Macroscopically, the incidence of colon tumors at a 24-26 week period of tumor induction was apparently lower in mice treated with both the DMH and dietary supplement (76%) than in those treated with DMH alone (100%). Histologically, microadenomas were induced predominantly in the anal half of the total colon, and large lymphoid aggregates were often associated with dysplastic crypts in the distal colon. Apoptotic cell masses were shed into the distended lumen of the involved crypts. The statistical analysis at a 15-week period of tumor induction indicated that the incidence of microadenomas per tumor-induced mouse was lowered significantly by use of the dietary supplement. From the present results, it is suggested that the intake of the dietary supplement inhibits the early development of colon adenomas, and the inhibition of microadenomas results in a reduction of subsequent polyp and tumor yield in the mouse colon. PMID- 11496320 TI - Efficacy of oral UFT plus leucovorin therapy for colon cancer with ovarian and multiple liver metastases: report of two cases. AB - Case 1: a patient was diagnosed as having ascending colon cancer with right ovarian metastasis, and underwent palliative right hemicolectomy plus oophorectomy. The tumor was a well-differentiated adenocarcinoma with right ovarian metastasis, and the disease was classified as stage IV. Oral chemotherapy with UFT plus LV was performed for about 3 years, and the patient is still being followed up with no recurrence at 5 years postoperatively. Case 2: a patient was diagnosed as having incomplete large bowel obstruction caused by ascending colon cancer, and underwent curative right hemicolectomy. The tumor was a moderately differentiated adenocarcinoma, and the disease was classified as stage II. Since multiple liver metastases developed at 3 months postoperatively, oral chemotherapy with UFT plus LV was started. Imaging studies showed the complete elimination of liver metastases after 2 months. Subsequently, liver metastasis recurred about 10 months later. The patient died of unrelated cerebral infarction at 2 years and 6 months postoperatively. PMID- 11496321 TI - DNA methylation status of hMLH1, p16(INK4a), and CDH1 is not associated with mRNA expression levels of DNA methyltransferase and DNA demethylase in gastric carcinomas. AB - DNA methyltransferase and DNA demethylase are enzymes potentially affecting promoter methylation status. We examined levels of DNA methyltransferase (DNMT1, DNMT3a, DNMT3b) and DNA demethylase (MBD2) mRNA expression by semi-quantitative RT-PCR. In addition, we examined promoter methylation status of hMLH1, p16(INK4a), and CDH1 by methylation-specific PCR since all three of these genes are reported to be hypermethylated in gastric carcinoma. MBD2 appeared to be down regulated in neoplasms. The levels of DNMT1, DNMT3a, DNMT3b, and MBD2 mRNA expression were not associated with either tumor stage or histologic type. Promoter hypermethylation of hMLH1, p16(INK4a), and CDH1 was detected in 5/20 (25%), 8/20 (40%) and 8/20 (40%) of gastric carcinomas, respectively. There was no clear relation between DNA methylation status of hMLH1, p16(INK4a), and CDH1 and the mRNA expression levels of DNMT1, DNMT3a, DNMT3b or MBD2. We divided the examined cases into two groups according to the number of hypermethylated genes. Cases with more than two hypermethylated genes comprised a hypermethylation group, and cases with no hypermethylation comprised a non-hypermethylation group. We found no group association for levels of DNMT1, DNMT3a, DNMT3b, and MBD2 mRNA expression. Our results suggest that the mRNA expression levels for pro methylating (DNMT1, DNMT3a, DNMT3b) and anti-methylating (MBD2) enzymes is not a critical determinate of tumor-specific promoter hypermethylation of hMLH1, p(16INK4a), or CDH1 in gastric carcinoma. PMID- 11496322 TI - Doenjang hexane fraction-induced G1 arrest is associated with the inhibition of pRB phosphorylation and induction of Cdk inhibitor p21 in human breast carcinoma MCF-7 cells. AB - Doenjang (fermented soy paste) is a Korean traditional fermented food. In previous studies, we demonstrated that Doenjang extracts exhibited anti-mutagenic and anti-carcinogenic activities. Here we investigated the effects of Doenjang hexane fraction (DHF) on cell cycle progression in the human breast carcinoma MCF 7 cells. Treatment of DHF to MCF-7 cells induced a G1 phase arrest of the cell cycle, which correlated with the accumulation of the hypophosphorylated form of the retinoblastoma protein (pRB) and enhanced association of pRB with the transcription factor E2F-1. After DHF treatment, the expression of D-type cyclins was decreased in a time-dependent manner, but DHF did not affect the levels of cyclin-dependent kinases (Cdks), cyclin E and cyclin A protein. However, the activity of Cdk2 and cyclin E-associated kinase was decreased in a time-dependent manner. The tumor suppressor p53 and Cdk inhibitor p21, a known downstream effector of the p53, and association of p21 with Cdk2 were markedly induced in DHF-treated cells. Taken together, the present results indicate that treatment of MCF-7 cells with DHF induces a significant inhibition of pRB phosphorylation and increase of Cdk inhibitor p21 that appear to be responsible for the observed G1 arrest. PMID- 11496323 TI - Plasma VEGF concentration can predict the tumor angiogenic capacity in non-small cell lung cancer. AB - We assessed the clinical utility of circulating angiogenic factors as a predictor for tumor angiogenesis in primary lung cancer. Circulating vascular endothelial growth factor (VEGF) and intratumoral VEGF were assessed by enzyme-linked immuno sorbent assay (ELISA). There was a significant increase in the mean value of both plasma and serum VEGF concentration in primary lung cancer patients (n=97) compared to those of healthy controls (n=59). There was a significant correlation between plasma VEGF levels and microvessel density (MVD), and also between plasma VEGF and intratumoral VEGF levels. Plasma VEGF in patients with lung cancer appears to be a useful indicator of tumor angiogenesis. PMID- 11496324 TI - Preoperative therapy with insufficient effectiveness is related to unfavorable prognosis of patients with esophageal squamous cell carcinoma. AB - The disadvantage brought by the preoperative therapy with insufficient effectiveness for the patients with esophageal carcinoma has not been previously investigated. The aim of the current study was to show that the prognosis of patients with advanced esophageal carcinoma treated with preoperative therapy with insufficient effectiveness may be rather more unfavorable than that of patients without preoperative therapy. The subjects were 406 patients with esophageal squamous cell carcinomas of depth reaching to muscularis propria or adventitia but not to neighboring structures, who had been treated with esophagectomy and reconstruction. Although the proportions of lymphatic invasion and venous invasion in patients treated with preoperative therapy of insufficient effect were lower than those of patients without preoperative therapy, the 1-, 3- and 5-year survival rates in the former (62.2%, 32.0% and 23.1%, respectively) was significantly lower than in the latter (78.5%, 57.3% and 39.7%, respectively, p=0.01). A preoperative therapy with insufficient effectiveness may bring a more unfavorable prognosis of patients with esophageal squamous cell carcinoma. PMID- 11496325 TI - Prognostic value of plasma transforming growth factor-beta in patients with glioblastoma multiforme. AB - We investigated whether the postoperative concentration of circulating transforming growth factor beta (TGF-beta) yields prognostic value in patients with glioblastoma multiforme (gbm). Blood was collected from 20 healthy volunteers and in 28 patients with mainly glioblastoma multiforme (gbm), both before radiotherapy, during and after 4 weeks of irradiation. Both latent and active TGF-beta were quantified directly in the blood plasma using a bioassay with mink lung epithelial cells transfected with a plasminogen activator inhibitor-1 promotor luciferase construct. The average plasma concentration of TGF-beta before radiotherapy for gbm patients was 26.2 ng/ml, which was significant higher than in normal controls (16.2 ng/ml, p=0.02). No correlation was found between TGF-beta and survival, nor between plasma TGF-beta and the diameter of the postoperative contrast-enhancing lesion. The pattern of plasma TGF-beta during radiotherapy did not correlate with the clinical course of patients, nor with the fractionation scheme. Plasma TGF-beta did not reveal a clinical useful prognostic value for gbm patients, which is partly due to the large variation in TGF-beta plasma levels between individual patients. PMID- 11496326 TI - Gemcitabine plus epirubicin in advanced pancreatic cancer: a phase II multicenter trial. AB - The aim of this phase II multicenter trial was to evaluate the activity of a novel combination of gemcitabine (GEM) and epirubicin (EPI) in advanced pancreatic cancer patients. Clinical benefit and response rate were the main efficacy end-points. From December 1997 to October 1999, 30 consecutive patients with measurable advanced pancreatic cancer were enrolled. Gemcitabine was administered intravenously in 30 min at a dose of 800 mg/m2 on days 1, 8, 15 followed by i.v. injection of epirubicin 25 mg/m(2); treatment was repeated every 28 days. With regard to clinical benefit response, 8/21 patients (38%) experienced significant palliation of tumor-related symptoms; the median symptom control time was 25 weeks. No complete responses were recorded while 6 patients achieved a partial remission, for an overall response rate of 20%; 10 patients (30%) had a stable disease and 14 (46%) had progressive disease. The median time to progression was 14 weeks. Median survival was 26 weeks, with 6 patients (20%) having long-term survival at 46 weeks. In general, chemotherapy was well tolerated; 9 patients (30%) suffered from WHO grade 3-4 haematological toxicity and 5 patients (16.6%) suffered from grade 3 non-haematological toxicity. In conclusion, the GEM plus EPI regimen represent a feasible approach for improvement of clinical benefit in advanced pancreatic cancer patients, but confirmatory investigations are required. PMID- 11496327 TI - Muscle wasting after radiotherapy in young and adult rats. AB - The impact of late muscular injury after radiotherapy on the well being of patients, particularly those treated at a relatively young age, is often underestimated. To extend the present knowledge, a detailed comparative study of the radiation response of skeletal muscles was performed. The hind limb of rats was irradiated at the age of 30-60 days (young), at approximately 200 days (adult) or at the age of >360 days (old), with single doses ranging from 15-60 Gy. Muscular wet weight, volume and tibia length was followed up to 12 months post-treatment. Radiation treatment resulted in muscular wasting, which was more severe at higher doses. The radiation response depended strongly on the age of the animal at the time of treatment. Radiation induced muscle wasting and stunted tibia growth were more prominent in young animals than in adult or in old rats. ED50 values based on normalized wet weight were 12.5 (11.5-13.6) Gy for young animals and 22.5 (21.8-23.3) Gy for adult animals. The gastrocnemius muscle of young animals appears to be more radiosensitive than the gastrocnemius muscle of adult or old animals. Furthermore, in young animals, severe wasting of the muscles may already occur after relative low radiation doses (single dose of 15 Gy). PMID- 11496328 TI - Expression of tumor-associated glycoprotein-72 (TAG-72) antigen in human prostatic adenocarcinomas. AB - Tumor-specific antigens are usually defined by monoclonal antibodies (MAbs) and can play critical roles in the diagnosis and therapy of carcinomas. Despite advances in the understanding of the molecular genetics of human prostate carcinomas, therapeutic approaches require that tumor-specific markers, preferably on the cell surface, should be defined. In this study, we examined the expression of an oncofetal antigen tumor-associated glycoprotein-72 (TAG-72) in prostatic adenocarcinomas with a Gleason grade of six or higher. Using a second generation MAb CC49 against TAG-72, immunoreactivity was detected in 88% (29/33) of the prostatic cancer tissues. Occasionally, the benign epithelium showed a very faint immunostaining but in most of the specimens, no reactivity was detected. Positive staining was present in the cytoplasm and the cell membrane of the malignant cells similar to reports on other cancer tissues. A weaker staining pattern of this antigen was seen in poorly differentiated areas. A significant negative correlation (r = -0.36, p < 0.05) was observed between TAG-72 antigen expression and Gleason grade. The TAG-72 antigen expression in prostatic adenocarcinomas may be used as a target for radioimmunotherapy by the multivalent single chain antibody CC49 constructs recently generated by our group. PMID- 11496329 TI - Expression of the c-erb-B2 and p53 protein in gallbladder carcinomas. AB - Gallbladder carcinoma is the most common malignancy of the biliary tract in Korea and known to be more common in East Asia and Latin America than in Europe and North America. However, their exact histopathological characteristics and carcinogenesis are not well-elucidated. A total of 71 cases of gallbladder carcinomas, two cases of gallbladder dysplasia and 20 cases of gallbladder adenoma were immunohistochemically studied to evaluate the expression of c-erb-B2 and p53 proteins in the light of their relationship with various prognostic factors. Thirty-three gallbladder carcinomas (46.5%) showed positive staining for c-erb-B2, but none of the dysplasia and adenoma were positive (p<0.05). c-erb-B2 was stained in the cell membrane of the cancer cells. Adjacent normal mucosa was negative for c-erb-B2 staining. Forty-eight gallbladder carcinomas (67.6%) showed positive staining in the nucleus for p53 protein. None of the cases with dysplasia, adenoma, and normal mucosa stained positive for p53 protein. There was no significant correlation between c-erb-B2 and p53 expression and age, gender, histological tumor grade, and tumor stage. In the multivariate analysis, tumor stage approached statistical significance (p=0.05), but c-erb-B2 and p53 expression was not significant (p=0.14 and p=0.29, respectively). The mean survival periods of the c-erb-B2 positive and negative groups were 26 months and 52 months, respectively (p=0.02). However, the mean survival periods of the p53 protein positive and negative patients were 34 months and 35 months, respectively (p=0.45). In conclusion, our results suggest that c-erb-B2 and p53 protein expression is strongly associated with neoplastic progression in gallbladder carcinomas, and that c-erb-B2 expression identifies patients with a worse prognosis. PMID- 11496330 TI - Loss of heterozygosity of nucleotide excision repair factors in sporadic oral squamous cell carcinoma using microdissected tissue. AB - The deficiencies of nucleotide excision repair (NER) factors are involved in rare genetic diseases such as xeroderma pigmentosum (XP) with increased risk of developing cancer on sun-exposed areas of the skin. However, the abnormality of NER factors in human sporadic carcinoma remains unclear. Loss of heterozygosity (LOH) analysis, using the microdissected tissues, for the XPA, XPB, XPC, XPD, XPE, XPF, XPG and the transcription-coupled repair factor, Cockayne syndrome B (CSB) revealed that NER factors were abnormal in 30.0% (3/10 cases) of oral squamous cell carcinomas. Furthermore, 10.0% of oral carcinomas exhibited LOH for NER factors without LOH for tumor suppressor genes such as p53, FHIT, APC, BRCA1, BRCA2 and DCC. These observations raise the possibility that alterations of NER factors may be involved in carcinogenesis in human oral squamous cell carcinoma. PMID- 11496331 TI - Cytomorphologic features characteristic of tumor stages of thymomas. AB - The cytologic findings of the tumor cells characteristic of the stages of thymomas were investigated to assess the invasiveness of the tumors. Forty-six patients with thymoma who underwent extensive thymectomy without pre-operative corticosteroid therapy were included in this study. The histologic subtypes included 18 round/oval, 20 mixed, and 8 spindle type. The stages of thymoma classified according to Masaoka's clinicopathological classification included 16 stage I, 20 stage II, 6 stage III, 2 stage IVa, and 2 stage IVb, and myasthenia gravis was recognized in 5 patients. Cytologic findings were retrospectively analyzed in the Papanicolaou-stained stamp smears obtained from the cut surfaces of thymoma specimens. Morphometry of the epithelial tumor cells using Cosmozone 1A was performed to evaluate the validity of our cytologic categories. Compared with the cytologic findings of stage I or II thymomas, those of epithelial tumor cells in stage III or IV more frequently showed necrotic background (50.0%-stage III or IV vs 11.1%-stage I or II, p=0.006), large clusters of epithelial tumor cells (70.0% vs 36.1%, p=0.055), marked nuclear enlargement (90.0% vs 52.7%, p=0.033), marked anisokaryosis (100% vs 52.7%, p=0.006), marked nuclear polymorphism (40.0% vs 5.5%, p=0.004), hyperchromasia (50.0% vs 11.4%, p=0.007) and prominent nucleoli (50.0% vs 16.6%, p=0.028) whereas no significant correlation was observed between cytologic findings and tumor volume. Morphometric studies of thymoma tumor cells revealed that the nuclear size (mean values, 78.8 microm(3)-stage III or IV vs 58.2 microm(3)-stage I or II), the coefficient of variation of the nuclear size (0.326 vs 0.282), and the nuclear rotundity (0.849 vs 0.858) differed significantly between the two categories (p<0.05). Our findings demonstrated that there were significant differences between the cytologic findings of epithelial tumor cells of stage I or II thymomas and those of stage III or IV thymomas, and that the cytologic findings of thymoma tumor cells appear to be useful for distinguishing between non invasive and invasive thymomas. PMID- 11496332 TI - The role of 18F-fluorodeoxyglucose positron emission tomography in cancer screening - a preliminary report. AB - Positron emission tomography (PET) with 18F-fluoro-2 deoxy-D-glucose (FDG) is a non-invasive method for surveying the whole body and detecting various malignancy. A total of 299 subjects underwent whole-body FDG PET studies in our PET center over an 18-month period. FDG PET accurately detected malignant tumors in 7 (2.34%) subjects. False positive FDG PET studies in 3 (1%) subjects and false negative FDG PET studies in 2 (0.6%) subjects were found. Because of the high cost of FDG PET examination, it might not be suitable as a cancer screening test for the general population. However, it is a valuable supplemented tool for routine check-up, particularly for those at high-risk of developing cancer. PMID- 11496333 TI - An intranodal morcellation technique employing loop electrosurgical excision procedure for large prolapsed pedunculated myomas. AB - For minimally invasive resection of large prolapsed pedunculated submucous myomas, improved techniques need to be established. An intranodal loop electrosurgical excision procedure (LEEP) was specifically developed for the morcellation of myoma tissue during vaginal myomectomy. Vaginal myomectomy was performed on a 28-year old woman with a large myoma occupying the small pelvis. A prolapsed pedunculated leiomyoma of the uterus 13 cm in maximal diameter was resected by intranodal morcellation of the myoma tissue employing LEEP. Intranodal morcellation technique utilizing LEEP is an effective method for the minimally invasive resection of large prolapsed pedunculated myomas. PMID- 11496334 TI - Role of iron in lymphoma-induction by ATP. AB - Iron complexed by ATP induces lymphomas in mouse organs other than the specific targets of the lympho-adenitis provoked by sodium ATP: lymph nodes, spleen and liver. The reduction of life spans and the production of substantial volumes of ascites, that are lacking in the case of sodium ATP, are an index of the degree of malignancy of the induced lymphomas. On the basis of the known characteristics of iron-ATP complex of cellular calcium homeostasis alteration, the mechanism of these phenomena is discussed. PMID- 11496335 TI - Expressions of c-erbB-2, epidermal growth factor receptor and pan-ras proto oncogenes in adenocarcinoma of the cervix: correlation with clinical prognosis. AB - The expressions of c-erbB-2, epidermal growth factor receptor (EGFR) and pan-ras in normal cervical glands (n=45), glandular dysplasia/adenocarcinoma in situ (GIN/ACIS) (n=32) and invasive cervical adenocarcinoma (n=78) were determined and correlated with clinical prognosis. The expressions of c-erbB-2, EGFR and pan-ras in GIN/ACIS lesions and invasive tumours were significantly higher than in normal glands (p<0.001), whereas there was no significant difference between expressions in GIN/ACIS lesions and invasive tumours, except for EGFR (p=0.016). Significantly more normal glands adjacent to adenocarcinoma showed moderate/strong expressions for EGFR than c-erbB-2 (p=0.007) whereas significantly more GIN/ACIS lesions showed moderate/strong expressions for c-erbB 2 than EGFR (p=0.008). No correlation was found between moderate/strong expressions for c-erbB-2, EGFR or pan-ras and stage at presentation (p=0.384, 0.056, 0.842 respectively) or with survival (p=0.58, 0.19, 0.26 respectively). In conclusion, EGFR is more important in inducing dysplastic change/malignant transformation whereas c-erbB-2 plays a more significant role in tumour progression and invasion. However, neither c-erbB-2, EGFR nor pan-ras carried any prognostic significance on patient survival. PMID- 11496336 TI - Immune reactivity during the growth of a murine lung adenocarcinoma: evaluation of paraneoplastic syndrome development. AB - The purpose of this study was to investigate if specific immune responses were present in mice bearing a lung adenocarcinoma that presents paraneoplastic syndromes during tumor evolution. Leukocytosis, mainly due to polymorphonuclear leukocytes, was found from day 15 of tumor growth. Delayed type hypersensitivity response and increased interleukin-6 (IL-6) serum levels were observed along tumor growth. Concomitant immunity, specific rejection of a second inoculum and in vitro specific cytotoxicity occurred at 20 days of implant. In advanced stages of tumor evolution impaired cytotoxicity, accompanied by a great increase of IL-6 in serum, were observed. Role of polymorphonuclear leukocytes and IL-6 overproduction as responsible for immune dysregulation and paraneoplastic syndromes are discussed. PMID- 11496337 TI - Pharmacokinetic and biochemical analysis in the treatment of weekly paclitaxel in relapsed breast cancer. AB - The mechanism(s) by which weekly paclitaxel exerted more therapeutic efficacy than the triweekly schedule in relapsed breast cancer is still unclear. To assess the rationale in therapeutic efficacy of weekly paclitaxel in relapsed breast cancer, pharmacokinetic and biochemical analyses were examined in terms of the mean peak plasma concentration at 0 min (Cmax), 30 min, and 24 h after finishing the infusion, and the extracellular domain of HER-2 in response to the treatment with paclitaxel. Twenty-five patients treated with weekly 1 h infusion of paclitaxel in the dose range from 40 mg/m(2) to 80 mg/m(2) were studied. Eleven patients responded to the treatment including 4 cases of complete response (CR) and 7 cases of partial response (PR), while 14 patients did not respond including 12 cases of no change (NC) and 2 cases of progressive disease (PD). The plasma concentration of paclitaxel and extracellular domain of HER-2 in the patients were measured by high-pressure liquid chromatography and enzyme immunoassay, respectively. The peak concentration (Cmax) and the other peaks at 30 min and 24 h in 10 patients including 3 cases of 40 mg/m(2), 3 cases of 60 mg/m(2) and 4 cases of 80 mg/m(2) in the weekly paclitaxel were compared in proportion to the increase of dose escalation, and compared to their tumor response. Further, the plasma levels of extracellular domain of HER-2 in 17 patients treated with the weekly paclitaxel were measured, and also compared to their tumor response. The mean Cmax treated with 40 mg/m(2), 60 mg/m(2) and 80 mg/m(2) in the weekly paclitaxel was 1.94, 2.18 and 1.54 microM, respectively. The dose escalation of paclitaxel and the dose intensity were not correlated with the increase of plasma concentration of paclitaxel nor with the tumor response. In contrast, the plasma level of extracellular domain of HER-2 in responders was higher than that of non responders in the weekly paclitaxel regimen(p=0.0512, Mann-Whitney's U-test). These results suggest that tumor response to the weekly 1 h infusion of paclitaxel was not associated with the plasma concentration and the dose intensity, rather the plasma level of extracellular domain of HER-2 protein may be a predictor of tumor response in the treatment of weekly paclitaxel in relapsed breast cancer. PMID- 11496339 TI - Editorial. PMID- 11496338 TI - Immunohistochemical study of Muc1, Muc2 and human gastric mucin in breast carcinoma: relationship with prognostic factors. AB - We investigated the expression of mucin core proteins, Muc1, Muc2, and human gastric mucin (HGM) immunohistochemically in 5 non-invasive, 62 invasive ductal carcinomas and 3 mucinous carcinomas of the breast and statistically examined the relationship with prognostic factors. Muc1 was expressed in almost all breast carcinomas and there was a reverse correlation with tumor size (r = -0.25). However, Muc1 was not significantly correlated with the other tumor characteristics such as lymphocytic infiltration, axillary lymph node metastasis, TNM and plasma levels of CA153. Muc2 and HGM were expressed in 5 cases each in invasive ductal carcinomas, 0 and 3 in non-invasive ductal carcinomas, and 2 and 1 in mucinous carcinomas, respectively. Muc 2 expression correlated with lymph node metastasis. HGM was found in the tumors without lymph node metastasis and lower levels of serum CA153. PMID- 11496340 TI - [Complications in the use of the halo fixator]. AB - 50 patients out of a total of 88 who underwent treatment using a halo-fixateur between 1987 and 1997 were available for a follow-up interview reviewing local complications and quality of life. Marginal discomfort was observed in 54% of patients, moderate complaints/transitory pain in 30%, prolonged and severe discomfort and pain in 16%. 62% of patients took no analgetics, 22% infrequently, and 16% continuously throughout treatment. In pin-tract infection local treatment was successful in 6 patients, in 3 patients the screws needed to be relocated. Complaints of dysphagia due to extended forced lordosis of the cervical spine could be corrected by adjusting the position of the halo ring in 3 out of 8 patients. 3 patients developed pressure sores which could be managed without surgical intervention. Proper fixation and placement of the pin-tracts are crucial in the application of the halo fixateur if complications are to be avoided. Superficial infections must be treated locally. If the infection persists immediate pin relocation and systemic antibiotic therapy have to be initiated. PMID- 11496341 TI - Trepanations from the early medieval period of southwestern Germany--indications, complications and outcome. AB - Of the 384 skulls from the early medieval period, 8 (2.1%) exhibited trepanations. Three skulls showed trepanations in the proximity of fractures. One case of infection after borrhole trepanation that resulted in death is identifiable. Seven skulls showed clear evidence of healing and survival after trepanation. The mortality and infection rates for trepanation were low during the 6th to 8th centuries in southwestern Germany. PMID- 11496342 TI - German neurosurgery in the first half of the 20th century. PMID- 11496343 TI - Hayari Miyake--a surgeon who built the bridge between Japan and Germany. PMID- 11496344 TI - Proliferative and apoptotic differences between alveolar rhabdomyosarcoma subtypes: a comparative study of tumors containing PAX3-FKHR or PAX7-FKHR gene fusions. AB - BACKGROUND: Most alveolar rhabdomyosarcomas (ARMS) have chromosome translocations and resultant gene fusion products. The more common translocation fuses the PAX3 and FKHR genes; patients who have PAX3-FKHR-positive ARMS have reduced event-free survival compared to patients with ARMS containing the less common translocation that fuses the PAX7 and FKHR genes. PROCEDURE: We examined histology, immunohistochemical markers of differentiation, and cell cycle characteristics of a panel of ARMS containing either PAX3-FKHR or PAX7-FKHR transcript to determine if these features differ between the ARMS subsets. RESULTS: Cell cycle parameters varied significantly: the number of nuclei that stained with either an immunohistochemical marker of proliferation (MIB1), or a TUNEL-based assay for apoptosis was significantly greater in tumors that expressed PAX3-FKHR compared to tumors that expressed PAX7-FKHR transcript. CONCLUSIONS: We conclude that compared to PAX7-FKHR-containing tumors, ARMS that contain PAX3-FKHR transcript have (1) increased cell proliferation, consistent with greater loss of cell cycle regulation, and (2) apoptosis that is increased but insufficient to prevent tumor formation. More marked cell cycle dysregulation may contribute to poorer prognosis for patients with ARMS that have PAX3-FKHR fusion. Med Pediatr Oncol 2001;37:83-89. PMID- 11496345 TI - Synovial sarcoma in children and adolescents: thirty three years of experience with multimodal therapy. AB - BACKGROUND: Synovial sarcoma (SS) is the most common type of non-rhabdomyosarcoma soft tissue sarcoma in childhood, with controversies about its prognosis and treatment. PROCEDURE: We reviewed medical records of 42 children and adolescents with SS treated at our institution between 1966 and 1999 to determine treatment results and assess prognostic factors. RESULTS: With a median follow-up duration of 7.8 years (range 0.2-22.4 years), 5-year progression free survival (PFS) and overall survival (OS) rates were 75.6% (95% Confidence Interval [CI] 62-89.2%) and 87.7% (95% CI 77.3-98.1%) respectively. Eleven patients were dead and four others had progressed but were alive without evidence of disease after further therapy. IRS grouping and tumor invasiveness were found to be significant prognostic indicators (P < 0.01 and = 0.02, respectively). Patients with initial gross total resection (IRS I and II) and non-invasive tumors (T1) were most likely to have prolonged PFS and OS. Patients with small tumors (< 5 cm) (P = 0.09) or with monophasic histology (P = 0.14) had better PFS and OS. CONCLUSIONS: Achieving a complete resection or gross total resection with microscopic residual disease is vital for survival of patients with localized SS. Patients with localized disease who received radiotherapy had improved local control. Chemotherapy did not seem to impact PFS or OS. Future large multi-institutional trials are needed to address whether post-operative chemotherapy is necessary for patients with localized, surgically removed tumors, whether radiotherapy is necessary for patients with completely resected tumors, and to ascertain the order of importance of all the candidate prognostic markers. Med Pediatr Oncol 2001;37:90-96. PMID- 11496346 TI - Prognostic significance of p80 and visceral involvement in childhood CD30 anaplastic large cell lymphoma (ALCL). AB - BACKGROUND: Between 1976 and 1998, CD30+anaplastic large cell lymphoma (ALCL) was diagnosed in 44 children (28 males, 16 females, age range 2.7-16.1 years, median 10). Up to 1993, 32 such children were treated according to a common chemotherapeutic protocol that stratified patients according to stage, without considering presenting features. Thereafter, patients presenting with visceral (lung, spleen, liver, gastro-intestinal tract) or mediastinal involvement were assigned to a high-risk treatment protocol with induction intensification. The generation of these two risk-groups was the result of a retrospective analysis of clinical risk factors for therapy failure as previously reported [Massimino M, Gasparini M, Giardini R, Ann Oncol 1995;6:915-920]. Considering the whole cohort of patients divided into group A-21/22 evaluable patients with visceral/mediastinal involvement, and group B-22 evaluable patients, with other ALCL location-disease-free survival (DFS) and survival (S) at 5 years were 57 and 58% for group A, and 83 and 100% (94% at 6 years) for group B, respectively. PROCEDURE: We tested 15/21 cases of group A, and 18/22 of group B for p80 immunoreactivity in order to investigate a possible correlation between ALCL locations and NPM-ALK expression. RESULTS: Thirteen of 15 specimens in group A and 17/18 in group B were positive for p80. CONCLUSIONS: It is impossible to conclude anything about p80 positivity based on a series of 33/44 patients with childhood ALCL, neither about over-all prognosis nor about the role of visceral involvement. In adults, NPM-ALK protein expression is a favourable prognostic factor. Med Pediatr Oncol 2001;37:97-102. PMID- 11496348 TI - Pattern and course of single-system disease in Langerhans cell histiocytosis data from the DAL-HX 83- and 90-study. AB - BACKGROUND: Single-system (SS) disease is the most common presentation in Langerhans cell histiocytosis (LCH) with a heterogenous clinical picture and course. Mostly bone and rarely skin or lymph nodes are involved. PROCEDURE: One hundred and seventy patients with SS-LCH were registered in the DAL-HX 83/90 studies. They were diagnosed according to uniform diagnostic criteria and followed by a standardised schedule. RESULTS: Single bone lesions were most common (68%), followed by multiple bone lesions (19%), isolated skin disease (11%), and isolated lymph node involvement (4 patients). In the detection of bone lesions radiographic skeletal survey proved to be superior to bone scan (97% vs. 82%). Treatment comprised surgery, irradiation and local instillation of steroids, and standardised chemotherapy for multifocal bone disease. After initial therapy 81% of the patients remained disease free. Reactivations restricted to the skeleton occurred in 18% of both unifocal and multifocal bone disease. Two skin patients had a chronic course. Fatality occurred only in one infant with skin disease who progressed to multi-system disease. Twenty-five percent of all patients developed permanent consequences, which were already present at diagnosis in about half of these patients and comprised mainly orthopedic problems related to lesional sites. Diabetes insipidus occurred in 3% and anterior pituitary dysfunction in 2% of the patients. CONCLUSIONS: The course in SS%LCH was benign. In bone disease reactivations remained restricted to the skeleton and did not influence survival. However, reactivations had an impact on morbidity, as permanent consequences were mostly related to the site of disease activity. Med Pediatr Oncol 2001;37:108-114. PMID- 11496347 TI - A phase II study of Homoharringtonine for the treatment of children with refractory or recurrent acute myelogenous leukemia: a pediatric oncology group study. AB - BACKGROUND: Homoharringtonine (HHT) is a plant alkaloid that is derived from a Chinese evergreen tree. Its mechanism of action is thought to be by inhibition of protein, DNA, and RNA synthesis by inhibition of chain initiation. The treatment of acute myelogenous leukemia (AML) in children, has been hampered by few new effective agents developed in the past 30 years. Significant numbers of children still die of this disease. The purpose of this study is to evaluate the efficacy and toxicity of HHT for the therapy of refractory AML in children. PROCEDURE: Patients entered the study and were treated with HHT 7mg/m(2)/day for 10 days. The cycles could be repeated every 21 days depending on recovery from myelosuppression. RESULTS: Thirty-seven patients entered the study, with twenty eight evaluable for response. Complete response was obtained in four and a partial response in one (response rate 5/28 = 18%). Significant toxicities included prolonged severe myelosuppression in all responsive patients and neuropathogenic pain in two patients. The median duration of response was 62 days with a range of 28-126 days. CONCLUSIONS: HHT has activity against chemotherapy resistant AML in children, with tolerable toxicity. This agent warrants further clinical evaluation in combination with other agents or perhaps biologic response modifiers which will hopefully lead to useful therapeutic options. Med Pediatr Oncol 2001;37:103-107. PMID- 11496349 TI - Delayed nausea and vomiting in children receiving antineoplastics. AB - BACKGROUND: The nature and prevalence of delayed antineoplastic-induced nausea and vomiting have not been well-described in children. This study describes the extent of delayed nausea and vomiting in children receiving antineoplastic agents as well as the drug therapies initiated in an attempt to prevent or manage it. PROCEDURE: All children receiving antineoplastics were eligible for study entry. The date and time of each emetic episode were recorded on each day antineoplastics were given and for 3 days thereafter. Nausea was self-assessed daily by children who were older than 3 years and were not developmentally delayed. Diet was also assessed daily. The emetic response, median nausea rating and median diet achieved were described. RESULTS: The emetic response of 124 children who received 174 antineoplastic cycles was evaluated. Most cycles (137/174;79%) were not associated with delayed vomiting. Cycles which included cisplatin, carboplatin, or cyclophosphamide; involved antineoplastic therapy given over 2 or more consecutive days; or were accompanied by vomiting during the acute phase were associated with a significantly higher incidence of delayed vomiting. Moderate to severe nausea was reported on 58% (267/459) of study days. No antiemetics were given on most study days (412/522;79%); nevertheless, most of the study days (381/412;93%) which were unaccompanied by antiemetic support during the delayed phase were completely free from vomiting. Antiemetics were most often given as single agents (ondansetron: 54 study days; dimenhydrinate: 17 study days; dexamethasone: 6 study days). Diet was largely unaffected during the study period. CONCLUSIONS: Antineoplastic-induced delayed nausea and vomiting may be less prevalent in children than in adults. Routine antiemetic administration during the delayed phase may not be warranted in all patients. Med Pediatr Oncol 2001;37:115-121. PMID- 11496350 TI - Scedosporium infection in immunocompromised patients: successful use of liposomal amphotericin B and itraconazole. AB - BACKGROUND: Invasive fungal disease is a major cause of death in immunocompromised patients. The filamentous fungus Scedosporium consists of two species, S. prolificans and S. apiospermum, which can cause infections in immunocompromised patients that are often fatal. A significant feature of this pathogen is its broad resistance to many antifungal agents, including amphotericin B. PROCEDURE AND RESULTS: Five cases of infection with Scedosporium spp. occurred in patients with haematologic malignancies over a 10-month period. Three patients with S. prolificans were severely immunosuppressed and neutropenic; two were in relapse and another was early post-matched unrelated bone marrow transplant. All three died despite treatment with various combinations of amphotericin B and itraconazole. Two patients who were less immunosuppressed and had a normal neutrophil count developed S. apiospermum infection. Both were successfully treated with liposomal amphotericin B and itraconazole. CONCLUSIONS: Disseminated infection in immunocompromised hosts with Scedosporium spp. is often fatal. However, in patients with a lesser degree of immunocompromise and particularly in those infected with the less virulent S. apiospermum, intensive antifungal therapy with liposomal amphotericin B and itraconazole may be associated with complete eradication of infection. Med Pediatr Oncol 2001;37:122-125. PMID- 11496351 TI - Fibromatoses in childhood: the desmoid/fibromatosis complex. PMID- 11496352 TI - Chronic recurrent multifocal osteomyelitis: a rare disorder presenting as multifocal bone lesions. PMID- 11496353 TI - False positive MIBG scan due to accessory spleen. PMID- 11496354 TI - Incidence of childhood cancer in Stavropol territory, Russia. PMID- 11496355 TI - Childhood acute lymphoblastic leukemia presenting with severe hepatic dysfunction. PMID- 11496356 TI - Noncutaneous varicella-zoster virus (VZV) infection with fatal liver failure in a child with acute lymphoblastic leukemia (ALL). PMID- 11496357 TI - Two secondary leukemias among 15 children given oral etoposide. PMID- 11496358 TI - Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) in a girl previously affected by acute lymphoblastic leukemia. PMID- 11496359 TI - Secondary acute myelocytic leukemia after successful chemotherapy with etoposide for Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis. PMID- 11496360 TI - Does a "false negative" MIBG scan predict a better outcome in neuroblastoma patients? PMID- 11496361 TI - Repeated achilles tendinitis after high dose methotrexate. PMID- 11496362 TI - Lack of association between the tryptophan hydroxylase gene A218C polymorphism and attention-deficit hyperactivity disorder in Chinese Han population. AB - Previous studies have suggested that the serotonergic (5-HT) system might be involved in the development of Attention-deficit hyperactivity disorder (ADHD). ADHD is frequently characterized by aggressive and impulsive behavior, a major symptom associated with reduction in serotonergic function. The tryptophan hydroxylase (TPH) gene is a reasonable candidate for ADHD because it encodes the rate-limiting enzyme in the process of 5-HT biosynthesis. In this study, we examined the relationship between the A218C polymorphism in TPH gene and ADHD. Sixty-nine ADHD patients and their biological parents were investigated. The A218C polymorphism in intron 7 of TPH gene was detected by PCR-RFLP method. No allele or genotype concerned with this A218C polymorphism was found to be associated with ADHD when analyzed with the haplotype relative risk method. Therefore, our data indicate that the TPH gene A218C polymorphism may not be a susceptibility factor of ADHD in the Chinese Han population. PMID- 11496363 TI - Novel and previously reported single-nucleotide polymorphisms in the human 5 HT(1B) receptor gene: no association with cocaine or alcohol abuse or dependence. AB - Evidence from animal self-administration and human genetics studies suggests that the serotonin(1B) (5-HT(1B)) receptor may be involved in modulating responses to cocaine or alcohol. We hypothesize that polymorphisms, including single nucleotide polymorphisms (SNPs), in the human 5-HT(1B) receptor gene, may be associated with individual differences in vulnerability to cocaine or alcohol abuse or dependence. A total of 210 subjects were studied, including individuals with a primary diagnosis (DSM-IV criteria) of cocaine abuse or dependence, alcohol abuse or dependence, and controls with no history of previous or current illicit drug or alcohol abuse or dependence. Genomic DNA samples were isolated from each individual. For 157 of the subjects, polymerase chain reaction (PCR) was used to amplify the entire coding region of the 5-HT(1B) receptor gene as well as parts of the 5' and 3' untranslated regions. PCR products were sequenced in forward and reverse directions on an automated sequencer. Amplified DNA from an additional 53 subjects was sequenced in the 5' untranslated region to gain additional data on the frequency of one identified SNP. Seven polymorphisms were identified: one novel SNP in the 5' untranslated region (UTR) of the gene (A 161T); one SNP not reported in any published scientific communication (but found to be recorded in GenBank) in the 3' UTR (A1180G); two novel dinucleotide deletions at positions - 184/- 183 and - 182/- 181; and three previously identified SNPs (T-261G, C129T, G861C). Data were stratified by ethnicity and pooled Relative Risk was calculated for combined alcohol abuse and dependence cases and controls, and also for combined cocaine abuse and dependence cases and controls. No significant differences between cases and controls were found. PMID- 11496364 TI - Lack of association between a functional polymorphism of the cytochrome P450 1A2 (CYP1A2) gene and tardive dyskinesia in schizophrenia. AB - Tardive dyskinesia (TD) is a common side effect of long-term medication with typical neuroleptics. TD presents itself by abnormal involuntary movements and may lead to a potentially disabling and chronic clinical course. A vast majority of patients suffering from schizophrenia are smokers. Smoking has been reported to induce the activity of the CYP1A2 enzyme, which is an established metabolic pathway within the disposition of antipsychotics. Recently, a C-->A genetic polymorphism in the first intron of the CYP1A2 gene was reported to influence CYP1A2 activity in smokers. Subsequently, a pharmacogenetic study in 85 U.S. patients with schizophrenia (44 smokers, 41 individuals with unknown smoking status) showed the C/C genotype to be associated with higher TD severity (measured by the Abnormal Involuntary Movement Scale, AIMS) than the A/C or A/A genotype. This finding prompted us to investigate whether this effect was also present in a larger German sample of 119 patients with schizophrenia (82 smokers, 37 individuals with unknown smoking status). However, we could not replicate the reported association. The median AIMS scores did not differ between individuals with the A/A, A/C, or C/C genotypes. In an additional analysis, we compared the genotypic and allelic distribution among individuals grouped according to the criteria established by Schooler and Kane [1982: Arch Gen Psychiatry 39:486-487] (persistent TD vs. absent TD). We did not observe a differential genotypic or allelic distribution between the two diagnostic groups. Thus, our results do not support the hypothesis that the C-->A polymorphism in the CYP1A2 gene is involved in the etiology of TD in the German population. PMID- 11496365 TI - Common polymorphisms in LRP and A2M do not affect genetic risk for Alzheimer disease in Northern Ireland. AB - Genetic variation in one of the major APOE receptors in the brain has been associated with increased risk for Alzheimer disease (AD). A C/T polymorphism in exon 3 and a tetranucleotide repeat polymorphism in the 5' region of the low density lipoprotein receptor-related protein gene have been reported to increase risk in some studies but these reports have not been universally replicated. In addition, genetic variation in another ligand of LRP, alpha-2 macroglobulin (A2M), has also been associated with increased AD risk. However, these reports also remain controversial. We have genotyped both LRP polymorphisms and two polymorphisms in the A2M gene in a large group of clinically well-defined AD cases and controls from the relatively genetically homogeneous Northern Ireland population. Comparison of genotype and allele frequencies for polymorphisms in LRP revealed no significant differences between cases and controls. Multiple logistic regression analysis performed to assess any possible interaction between LRP and APOE revealed little evidence for genetic interaction despite the obvious biological interaction. Genotype and allele comparisons between the groups for the A2M polymorphisms also gave no evidence that either polymorphism increased risk for disease. The results from this study indicate that polymorphisms in LRP and A2M are not associated with increased risk for AD in Northern Ireland. PMID- 11496366 TI - Evidence for a susceptibility locus on chromosome 6q influencing phonological coding dyslexia. AB - A linkage study of 96 dyslexia families containing at least two affected siblings (totaling 877 individuals) has found evidence for a dyslexia susceptibility gene on chromosome 6q11.2-q12 (assigned the name DYX4). Using a qualitative phonological coding dyslexia (PCD) phenotype (affected, unaffected, or uncertain diagnoses), two-point parametric analyses found highly suggestive evidence for linkage between PCD and markers D6S254, D6S965, D6S280, and D6S251 (LOD(max) scores = 2.4 to 2.8) across an 11 cM region. Multipoint parametric analysis supported linkage of PCD to this region (peak HLOD = 1.6), as did multipoint nonparametric linkage analysis (P = 0.012). Quantitative trait linkage analyses of four reading measures (phonological awareness, phonological coding, spelling, and rapid automatized naming speed) also provided evidence for a dyslexia susceptibility locus on chromosome 6q. Using a variance-component approach, analysis of phonological coding and spelling measures resulted in peak LOD scores at D6S965 of 2.1 and 3.3, respectively, under 2 degrees of freedom. Furthermore, multipoint nonparametric quantitative trait sibpair analyses suggested linkage between the 6q region and phonological awareness, phonological coding, and spelling (P = 0.018, 0.017, 0.0005, respectively, for unweighted sibpairs < 18 years of age). Although conventional significance thresholds were not reached in the linkage analyses, the chromosome 6q11.2-q12 region clearly warrants investigation in other dyslexia family samples to attempt replication and confirmation of a dyslexia susceptibility gene in this region. PMID- 11496367 TI - Tryptophan hydroxylase gene variant and smoking behavior. AB - Approximately 50% of the variance in smoking behavior is attributable to genetic factors. Genes in the serotonin system are plausible candidates because of serotonin's role in mood regulation. The present study examined the association of smoking behavior with a polymorphism in the TPH gene, which codes for a rate limiting enzyme in the biosynthesis of serotonin. A polymorphism in intron 7 has been linked with a variety of traits involving poor impulse control. Participants in this study were 249 Caucasian smokers and 202 nonsmokers recruited through newspaper advertisements. Smokers completed smoking history and nicotine dependence assessments. The overall frequencies of the A- and C-allele were 42% and 58%, respectively. There was no association of TPH alleles with smoking status. However, case series analysis indicated that individuals with the A/A genotype started smoking at age 15.6 years, compared with 17.3 years among smokers with other genotypes. This association was significant in a multivariate regression model controlling for age, education, body mass index (BMI), alcohol use, and medication use. This finding is consistent with previous studies relating the A-allele to impulsive behavior and suggests that it may predispose to early smoking initiation. Future family-based studies are needed to confirm this finding. Published 2001 Wiley-Liss, Inc. PMID- 11496368 TI - PTEN mutation in a family with Cowden syndrome and autism. AB - We report on a mother and son with Cowden syndrome and a PTEN mutation. The boy also exhibits autistic behavior and mental retardation, while his mother has a normal intelligence and social interaction pattern. We review the scanty literature data on the association of Cowden syndrome and autism and emphasize that the association of progressive macrocephaly and pervasive developmental disorder seems to be an indication for screening for PTEN mutations. PMID- 11496369 TI - No association between a promoter dopamine D(4) receptor gene variant and schizophrenia. AB - The dopamine D(4) receptor has been implicated in the pathogenesis of schizophrenia. An association between a putative functional promoter polymorphism (-521C/T) in the dopamine D(4) receptor gene (DRD4) and schizophrenia was recently reported. In the present study, patients with schizophrenia (n = 132) and control subjects (n = 388) were analyzed with respect to the DRD4 - 521C/T polymorphism. No significant case control differences emerged. The present results do not support a major role for DRD4 in the etiology of schizophrenia among Caucasians from Sweden. PMID- 11496370 TI - Association study of a promoter polymorphism of UFD1L gene with schizophrenia. AB - Schizophrenia or schizoaffective disorders are often found in patients affected by DiGeorge/velo-cardio-facial syndrome (DGS/VCFS) as a result of hemizygosity of chromosome 22q11.2. We evaluated the UFD1L gene, mapping within the DGS/VCFS region, as a potential candidate for schizophrenia susceptibility. UFD1L encodes for the ubiquitin fusion degradation 1 protein, which is expressed in the medial telencephalon during mouse development. Using case control, simplex families (trios), and functional studies, we provided evidence for association between schizophrenia and a single nucleotide functional polymorphism, -277A/G, located within the noncoding region upstream the first exon of the UFD1L gene. The results are supportive of UFD1L involvement in the neurodevelopmental origin of schizophrenia and contribute in delineating etiological and pathogenetic mechanism of the schizophrenia subtype related to 22q11.2 deletion syndrome. PMID- 11496371 TI - Genetic variation of the human glycine receptor subunit genes GLRA3 and GLRB and susceptibility to idiopathic generalized epilepsies. AB - Alterations of glycine receptor alpha1 and beta subunit genes have been associated with hypertonic motor disorders in both mice and humans. Mutations in genes encoding other ligand- and voltage-gated ion channels have been identified in rare monogenic forms of idiopathic generalized epilepsies (IGE). We tested the hypothesis that allelic variants of the glycine receptor subunit genes, GLRA3 and GLRB, both localized on chromosome 4q, confer susceptibility to common subtypes of IGE. Mutation screening was carried out in index patients of 14 IGE families. No pathogenic mutation was found, but two intronic polymorphisms were detected in the GLRB gene, and four intronic, three exonic, and one 3'-UTR polymorphisms were identified for the GLRA3 gene. Subsequent screening for exonic and 3'-UTR polymorphisms in GLRA3 showed no statistical difference between a group of sporadic IGE patients (n = 104) and a control group (n = 141). The genotype frequencies for exonic and 3'-UTR polymorphisms in GLRA3 showed no statistically significant difference between IGE patients (n = 104) and an ethnically matched control group (n = 141). Thus, no association between IGE and alterations in GLRA3 or GLRB genes could be detected, indicating that both genes do not play a major causative role in the epileptogenesis of common IGE subtypes. Still, these novel single nucleotide polymorphisms may be useful markers for candidate gene analyses of other disorders. PMID- 11496372 TI - Incorporating language phenotypes strengthens evidence of linkage to autism. AB - We investigated the effect of incorporating information about proband and parental structural language phenotypes into linkage analyses in the two regions for which we found the highest signals in our first-stage affected sibling pair genome screen: chromosomes 13q and 7q. We were particularly interested in following up on our chromosome 7q finding in light of two prior reports of linkage of this region to developmental language disorder, since one of the diagnostic criteria for autism is absent or abnormal language development. We hypothesized that if the language phenotype were genetically relevant to linkage at the chromosome 7q locus, then incorporating parents phenotypes would increase the signal at that locus, and most of the signal would originate from the subset of families in which both probands had severe language delay. The results support these hypotheses. The linkage signals we obtained on chromosome 7q as well as at least one signal on chromosome 13q are mainly attributable to the subgroup of families in which both probands had language delay. This became apparent only when the parents' history of language-related difficulties was also incorporated into the analyses. Although based on our data, we were not able to distinguish between epistasis or heterogeneity models, we tentatively concluded that there may be more than one autism susceptibility locus related to language development. PMID- 11496373 TI - Linkage genome scan for loci predisposing to panic disorder or agoraphobia. AB - We conducted a 10 cM linkage genome scan in a set of 20 American pedigrees (153 subjects), ascertained through probands with panic disorder (PD). Several anxiety disorders segregate in these families; they were diagnosed on the basis of Schedule for Affective Disorders and Schizophrenia interview. In this article, we describe results for panic disorder and agoraphobia, which are closely related, common, heritable anxiety disorders. This is the first complete linkage genome scan for agoraphobia and the third for PD. A total of 407 markers (389 autosomal, 18 X chromosome) were genotyped. Multipoint LOD score and NPL analysis were completed using GENEHUNTER2. For PD, two genomic regions meet criteria for suggestive linkage. One of these regions is on chromosome 1 (LOD score = 2.04). This region coincides with a region that generated a LOD score of 1.1 in a previous genome scan by Crowe et al. [2001: Am J Med Genet (Neuropsychiatr Genet) 105:105-109]. The other (LOD score = 2.01) is located on chromosome 11p and occurs at marker CCKBR, one of eight candidate genes examined. For agoraphobia, the most promising potential linkage was on chromosome 3 (NPL score = 2.75; P = 0.005). This was accounted for primarily by a single family that by itself generated an NPL score of 10.01 (P = 0.0039) and a LOD score of 2.10. These results provide initial evidence for a genetic locus on chromosome 3 that contributes to risk for agoraphobia. They also support suggestive linkage to two risk loci for panic disorder. Additional potential loci were identified with lesser statistical support; several of these were consistent with previously reported panic disorder linkage results. Overall, the results presented here suggest that PD and agoraphobia are complex traits that share some, but not all, of their susceptibility loci. Published 2001 Wiley-Liss, Inc. PMID- 11496374 TI - [Double-blind clinical trial of aspirin and Fenbufen in rheumatoid arthritis]. PMID- 11496375 TI - [Double-blind comparative study of bufexamac and triamcinolone acetonide given by local injection in abarticular rheumatism or osteoarthritis]. PMID- 11496376 TI - [Clinical experiences with an anti-arthritic agent. The use of a "heparinoid" in long-term cases of osteoarthritis]. PMID- 11496377 TI - [Dupuytren's disease. Treatment by local injections with hyaluronidase using a Dermojet]. PMID- 11496378 TI - [Fenbufen in osteoarthritis]. PMID- 11496379 TI - [Post-radiotherapy humeral head necrosis in breast cancer]. PMID- 11496380 TI - [Physico-chemical parameters of normal bone, osteoporosis, and osteomalacia]. PMID- 11496381 TI - [Vertebral body disappearance. Radiographic presentation]. PMID- 11496382 TI - [Risk of developing ankylosing spondylitis in "healthy" persons positive for HLA B27 antigen]. PMID- 11496383 TI - [Evaluation of bone scintigraphy with pyrophosphate and fibrinogen labeled with technetium 99m in rheumatoid arthritis and other arthropathies]. PMID- 11496384 TI - [The 9th European Congress of Rheumatology to insist on adherence to total rehabilitation]. PMID- 11496385 TI - Isotretinoin (roaccutane) usage--a South African consensus guideline. National Dermatology Working Group. AB - OBJECTIVE: This guideline has been developed in order to recommend the correct and safe usage of isotretinoin (Roaccutane) in the treatment of acne and related conditions. Aspects such as correct dosage related to the patient's weight and the minimal requirement in relation to blood tests and physician visits have been addressed. OPTIONS: Isotretinoin is known to be the treatment of choice for severe acne. Familiarity with the drug has lowered the threshold for its use in lesser degrees of acne, particularly when scarring is occurring and other options outlined in the guideline have failed or are either unsuitable or not tolerated. OUTCOMES: Severe acne causes permanent physical damage and frequently equally severe psychological damage. The correct use of isotretinoin increases the chance of complete cure, thus maximising the chance of meeting the patient's expectations. EVIDENCE: Studies have shown that the patient may expect the chance of permanent cure to be between 60% and 80% at a dose ranging from 0.5 to 1 mg/kg body weight per day for a period of 16-20 weeks. Recent evidence suggests that a cumulative dose of 120 mg/kg body weight during the course at a daily dose of 0.75 mg/kg body weight or greater may be more important than the duration of treatment. VALUES: The National Dermatology Working Group consisting of 11 consultant dermatologists, all with considerable experience in the use of isotretinoin therapy for acne, using the current literature as back-up, reached consensus on the contents of the document. The document was presented to the Congress of the South African Dermatology Society in May 1997. It was subsequently sent to all registered dermatologists in South Africa for comment. BENEFITS, HARM AND COSTS: Correct use of isotretinoin will often effect cure and at worst considerably improve acne. Side-effects due to hypervitaminosis A, while almost inevitable, are uncomfortable rather than dangerous. The potential for teratogenicity is stressed. Isotretinoin is an extremely expensive drug, but studies have shown that it is cost-effective when compared with prolonged treatment with other modalities in what is usually a chronic condition. RECOMMENDATIONS: The optimal dosage recommended is a cumulative dose of 120 mg/kg body weight with a daily dose as close to 1 mg/kg body weight as possible. Isotretinoin is used for severe acne, lesser degrees of acne where scarring is seen, and in patients who do not respond to or cannot tolerate other treatment modalities. VALIDATION: This guideline is similar to those recommended by other groups outside South Africa. It has been scrutinised by all the practising dermatologists in the country. It is endorsed by the Medical Association of South Africa. PMID- 11496386 TI - Eating disorders. AB - An eating disorder is many things-deadly, addictive, uncontrollable, and terrifying. Most people have a general knowledge of anorexia and bulimia, but few really understand the complexity of eating disorders. PMID- 11496387 TI - Oral pathology. Case of the quarter. Adenoid cystic carcinoma. PMID- 11496388 TI - Oklahoma dental Medicaid. A need for action. AB - Dental Medicaid in the State of Oklahoma must be made to work. But it will not work unless enough of us get involved and lead health officials, and legislators into the serious work that can turn Medicaid's promise into performance and a Medicaid card into meaningful dental care. Ironically, Medicaid's "EPSDT" (Early Periodic Screening, Diagnosis and Treatment) authorizes a high scope of dental coverage for the child. But trying to access care for the Medicaid eligible child is entirely another matter. Only a small number of Oklahoma dentists treat the quarter of a million Medicaid eligible persons in Oklahoma. Why is this so? PMID- 11496389 TI - Managing the public perception of dentistry. PMID- 11496390 TI - Scheduling: 3 essentials. Productivity. Profitability. Stress control. PMID- 11496391 TI - Management of the child dental patient. AB - The ultimate goals of behavior management for children receiving dental treatment are to achieve optimum oral health, instill a positive attitude in children and their parents regarding oral health, instill confidence in children regarding their ability to undergo dental treatment successfully and complete treatment in a safe and expeditious manner with the least invasive technique possible. These goals can only be achieved by using methods of behavior management that encourage positive communication and reduce anxiety for the patient, parent, and staff while educating children and parents about the importance of oral health. All behavior management techniques must consider the risks and benefits to the child, his or her parents, as well as the dental staff. PMID- 11496392 TI - [Evaluation of usefulness of urinary nuclear matrix protein 22 (NMP22) in the detection of urothelial transitional cell carcinoma]. AB - We evaluated the usefulness of urinary nuclear matrix protein 22 (NMP22) compared to urinary cytology in the detection of urothelial transitional cell carcinoma (TCC). Between July 1999 and March 2000, 227 patients complaining of microscopic or gross hematuria were analyzed. Twenty-four patients (10.6%) had urothelial TCC. The urinary NMP22 level was significantly higher in the patients with urinary TCC compared to the other patients. The sensitivity and specificity of the results obtained with urinary NMP22 were 58.3% and 84.2%, respectively, and those obtained by urinary cytology were 45.8% and 98.0%, respectively. False positive results were obtained with urinary NMP22 in the patients with urinary diversion using intestine, bladder invasion from other cancers, urinary tract infection, and urolithiasis. The urinary NMP22 level was significantly associated with tumor stage, suggesting its usefulness for detection of urothelial TCC. However, although urinary NMP22 showed equal sensitivity for the detection of TCC, it was not superior to urinary cytology. PMID- 11496393 TI - [Diagnosis of prostate capsular invasion by pelvic magnetic resonance imaging and serum level of prostate specific antigen]. AB - Transrectal ultrasonography (TRUS), computed tomography (CT), and magnetic resonance imaging (MRI) are employed to diagnose the clinical stage of prostate cancer. However, several cases are diagnosed as pathological stage pT3 after total prostatectomy. We investigated the accuracy of the evaluation of pathologic capsular penetration by preoperative pelvic MRI and preoperative serum PSA level and capsular penetration. The diagnostic accuracy of capsular penetration by MRI was 63.3%. On the other hand, the diagnostic accuracy of capsular penetration by preoperative PSA was 89.7% when its cut off value was 17 ng/ml. We conclude that preoperative serum PSA level could be more useful to diagnose accurately stage of prostate cancer than pelvic MRI. PMID- 11496394 TI - Clinical study of testicular germ cell tumors. AB - A clinical statistical analysis on 65 patients with 68 testicular germ cell tumors was performed. Thirty-six testes (53.7%) had seminomas and the remainder non-seminomatous germ cell testicular tumors (NSGCTTs). Of the seminomas, 31 (88.6%) were in stage I and the others showed distant metastases at presentation. Of the 32 NSGCTTs, 22 (68.8%) were in stage I. The average ages of the patients with seminomas and NSGCTTs were 40.4 and 29.9 years, respectively. Thirty-nine patients (60.0%) had tumors on the right side, 23 (35.4%) on the left and 3 (4.6%) in both testes. Five patients had a past history of cryptorchidism. Chief complaints in 49 patients (73.1%) were a painless scrotal mass. The interval from clinical onset to presentation was longer in seminoma patients than in NSGCTT patients (10.9 months on average versus 3.4 months). Immunosuppressive acidic protein (IAP) was a useful diagnostic tumor marker as well as alpha-feto protein (AFP), beta-human chorionic gonadotropin (beta-hCG) and lactic dehydrogenase (LDH). We adopted a surveillance policy in more than half of the stage I patients and obtained acceptable results. In the remaining cases, therapies including combination chemotherapy, radiation and salvage operation were performed after orchiectomy. The three-year survival rate was 98.0, 100.0 and 26.7%, for stage I, II and III patients respectively. PMID- 11496395 TI - [Peripheral blood stem cell harvest for patients with germ cell tumors]. AB - From January 1996 to December 1999, fifteen patients with germ cell tumors underwent peripheral blood stem cell harvest during 15 courses of bleomycin, etoposide, cisplatin (BEP), 4 courses of etoposide, ifosfamide, cisplatin (VIP) and 3 courses of high-dose etoposide mobilization at Nagoya University Hospital. We performed 29 aphereses during BEP, eight during VIP, and six during high-dose etoposide. Although we were able to harvest 4.4 x 10(6)/kg of median CD34 positive cells per apheresis during BEP, the number of stem cells (more than 4 x 10(6)/kg of CD34 positive cells), which are needed for tandem high-dose chemotherapy, could not be obtained during four courses of BEP. For three patients in whom white blood cell counts at nadir were 2,000/microL or more, however, the required number of CD34 positive cells were harvested. VIP provided only 1.7 x 10(6)/kg of median CD34 positive cells per apheresis, while, 7.3 x 10(6)/kg of CD34 positive cells were harvested during high-dose etoposide mobilization. The dose of G-CSF was a significant factor for the number of CD34 positive cells harvested during BEP (p = 0.02); however, there might be some relationship between the harvest and the number of the peripheral white blood cells on the day of apheresis (p = 0.08), the day to start G-CSF (p = 0.13), or the day to initiate apheresis (p = 0.27). Based on our experience, it is recommended that 5 micrograms/kg of G-CSF should be started from the 14th or 15th day of BEP until the last apheresis and that aphereses should be performed between the 19th and 21st day, especially at the days when the peripheral white blood cell count increases beyond 10,000/microL. PMID- 11496396 TI - [Multiple primary cancers limited to the urological field]. AB - We analyzed the clinical features of multiple primary cancers arising from the urogenital organs. Between January 1980 and December 1999, 300 patients with renal cell carcinoma (RCC), 661 patients with urothelial carcinoma (bladder cancer and renal pelvic-ureteral cancer) (TCC) and 391 patients with prostate cancer (PC) were treated at our hospital. Of these patients, 20 patients had double genitourinary cancers. The double cancers consisted of RCC and TCC in 1 case, RCC and PC in 6 cases, and TCC and PC in 13 cases. Seven cases had synchronous tumors. The average interval in the metachronous cases was 68 (range: 12-209) months. The age at diagnosis of the second cancer was 68-94 (mean: 77.6) years old. The follow-up period ranged from 4-168 (mean: 38) months; Six patients are alive with no evidence of disease and 6 patients died of cancer. Even when limited to the urological section, the frequency of multiple primary cancers is increasing. PMID- 11496397 TI - [Primary small cell carcinoma of the kidney: a case report]. AB - A 43-year-old male visited our hospital with the complaint of right flank colicky pain. Computed tomographic (CT)-scan and angiography showed large renal tumor with liver invasion and tumor thrombosis in the vena cava. Multiple lung and bone tumors were also recognized. Percutaneous biopsy of the renal tumor revealed small cell carcinoma. Multiple lung masses were diagnosed as metastatic tumors according to the results of bronchoscopic biopsy. Chemotherapy including cisplatinum and etoposide was performed without success. He died 6 months after the diagnosis. Autopsy specimen revealed primary small cell carcinoma of the right kidney. To our knowledge, this is the seventh case as primary renal small cell carcinoma in the world literature. PMID- 11496398 TI - [Long-term experience with renal transplantation in systemic amyloidosis: a case report]. AB - Renal transplantation was performed on a 39-year old woman with secondary amyloidosis due to rheumatoid arthritis. She remains alive and renal function has been maintained satistfactorily with the exception of proteinuria ten years after transplantation. Recent renal biopsy showed no amyloid recurrence, but the presence of chronic rejection reaction and mild cyclosporin arteriolopathy. Symptoms related to systemic amyloidosis and rheumatoid arthritis improved after transplantation. Renal transplantation is the recommended therapy for the type AA systemic amyloidosis. This is the second report of long-term experience with renal transplantation in systemic amyloidosis in Japan. PMID- 11496399 TI - [Primary adenocarcinoma of the renal pelvis with a pyeloduodenal fistula: a case report]. AB - A 62-year-old woman visited our hospital complaining of fever and right flank pain. On excretory pyelography the right kidney containing three renal stones was not visualized. Retrograde pyelography revealed an irregular filling defect in the right renal pelvis. Computed tomography revealed renal stones and a tumor mass in the right renal pelvis. From these findings, the pelvic tumor of the right kidney complicated by renal stones was diagnosed. Right nephrectomy was performed. Because a fistula between the renal pelvis and the second portion of the duodenum was found in the operation, partial resection of the duodenum was also performed. Pathological diagnosis was adenocarcinoma of the renal pelvis and pyeloduodenal fistula due to chronic pyelonephritis. Postoperative chemotherapy was not given. This is the first case report of adinocarcinoma of the renal pelvis coexisting with a pyeloduodenal fistula. PMID- 11496400 TI - [A case of congenital hydronephrosis suffering from rupture of the renal pelvis due to trauma]. AB - A 19-year-old male patient was admitted with the chief complaint of left abdominal pain. After receiving a mild punch in the abdomen during boxing exercises, he had severe abdominal pain and was brought to an emergency room. Since abdominal CT scanning revealed the retention of massive fluid in the retroperitoneum, hydronephrotic rupture due to the trauma was diagnosed and nephrectomy was performed. The removed kidney was filled as a result of urinary retention, and congenital hydronephrosis accompanied by the ureteropelvic junction obstruction was macroscopically and pathohistologically diagnosed. Postoperative course was favorable and the patient was discharged on the 10th hospital day. PMID- 11496401 TI - [Bladder carcinoma producing granulocyte colony-stimulating factor (G-CSF): a case report]. AB - An 84-year-old male was admitted to our hospital with complaints of leg edema and general fatigue. He had undergone transurethral resection of transitional cell bladder cancer, grade 3, pT1b, one year previously. The computed tomographic scan revealed a hypoattenuating bladder tumor, protruding extravesically. Laboratory examination showed remarkable leukocytosis of 46,900/mm3 in the peripheral blood and high value of granulocyte colony-stimulating factor (G-CSF) 226 pg/ml (normal: less than 30 pg/ml). The resection of the tumor (partial cystectomy) was performed. The histological diagnosis was transitional cell carcinoma, grade 3. The production of G-CSF was confirmed by immunohistochemical examination in the recurrent tumor and the surgical sample from the transurethral resection. The leukocyte count in the peripheral blood decreased to the normal range after surgery. But leukocytosis recurred one month postoperatively and the computed tomographic scan revealed intrapelvic tumor recurrence. He died due to drastic progression of recurrent tumor at three months postoperatively. PMID- 11496402 TI - [Primary squamous cell carcinoma of the prostate forming a rectourethral fistula: a case report]. AB - Eleven cases of squamous cell carcinoma of the prostate have been previously reported in the Japanese medical literature. Patients with this type of carcinoma have dismal prognoses. Herein, we report a case in a 61-year-old man who was admitted to our hospital and whose chief complaints were dysuria and high fever. Pyuria, bacteriuria, and the results of a digital rectal examination suggested acute prostatitis. Despite the administration of antibiotics, the dysuria did not improve. A transrectal needle biopsy was performed, and histology of the tissue showed squamous cell carcinoma. After chemotherapy using methotrexate, pirarubicin, and cisplatin, total pelvic excentration was performed. At 12 months after the operation, the patient is alive with no evidence of the disease. PMID- 11496403 TI - [Undescended testicular tumor found by torsion of the testis: a case report]. AB - A 37-year-old man was admitted with a painful mass in his left inguinal region. He had an undescended testis on the left side. Six months earlier, he had noted that his left inguinal testis was larger, and he had suddenly developed pain in the left inguinal region. The levels of AFP, hCG beta and LDH were normal. We diagnosed a left undescended testicular tumor and torsion of the left testis. Left inguinal high orchiectomy showed a torsion of the left testis and histopathological examination of the specimen revealed seminoma. PMID- 11496404 TI - [Rupture of intrascrotal epidermoid cyst complicated by bacterial infection: a case report]. AB - A 63-year-old male visited our hospital with the chief complaint of right scrotal pain. The right scrotum was swollen to the size of a small egg, and its skin was reddish. The mass was palpable independent of the right testis and epididymis. We diagnosed an intrascrotal abscess. The pus spontaneously issued from the scrotal mass. Sequentially, the abscess was extracted under spinal anesthesia. Membrane like tissue assumed as the abscess wall was removed. Histologically, the abscess wall was composed of epidermal structure with epidermal keratinization, and horny material was found inside the wall. In the scrotal epidermis overlying the abscess, infiltration of neutrophils, lymphocytes, and multinucleated giant cells were observed. Anaerobic bacteria were detected in the pus of the abscess. Consequently, we diagnosed this case as rupture of an intrascrotal epidermoid cyst complicated by bacterial infection. PMID- 11496405 TI - [A case of fibrous pseudotumor in the scrotum]. AB - A 65-year-old man presented with a complaint of the swelling of his right scrotum. Ultrasonogram indicated a tumor in the right scrotum, but it was not apparent whether the tumor arose from the right testis or not. Right high orchiectomy was performed due to the suspicion of malignancy. The tumor arose from the right epididymis and spermatic cord. Histological examination revealed that the tumor was a fibrous pseudotumor in the scrotum. PMID- 11496406 TI - [Fact-findings from comparison between Japan and U.S. in terms of visiting rate and oral health]. PMID- 11496407 TI - [Expression of N-deacetylase/N-sulfotransferase (NDST) during early rat embryogenesis]. AB - Fibroblast growth factors (FGFs) are known to be required for embryonic growth, formation, and shaping of many tissues and organs during embryogenesis. Biochemical studies have indicated that heparan sulfate proteoglycans, which are present on most cell surfaces and in extracellular matrices, play essential roles in FGF binding to its receptors and subsequent signal transduction. N deacetylase/N-sulfotransferases (NDSTs), which catalyze the first modifying step in the biosynthesis of heparan sulfate, are essential enzymes for the biosynthesis of heparan sulfate. The critical roles of heparan sulfate proteoglycan in developmental processes and specific signaling pathways have been illustrated by the identification of mutations in enzymes involved in the biosynthesis of heparan sulfate in Drosophila and mice. However, null mutation analyses of NDST-1 and NDST-2 have not clarified the embryonic regulation of morphogenesis by NDSTs. In the present study, we examined the temporal and spatial expression of NDST-1, and the temporal expression of NDST-2, in early rat embryogenesis, to investigate the role of NDSTs during embryogenesis. The expression pattern of rat NDST-1 appears to colocalize with certain FGFs and FGF receptors, suggesting possible roles of NDST in embryogenesis. PMID- 11496408 TI - [A study on improving tongue functions of open-bite children at the mixed dentition period--modifications of a removable habit-breaker appliance and their sonographic analysis]. AB - In open-bite cases at developing dental stages, oral morphologies have been thought to have a close relationship to tongue movement. The aim of this study was to design a new type of removable habit-breaker, and to examine its effects on tongue movement compared with the conventional habit-breaker. The examinees in the mixed dentition period, who had anterior open bite with tongue thrust, consisted of two groups: 10 children who had been treated with conventional habit breakers with tongue cribs (the upper group), and 10 children treated with the modified habit-breakers (the lower group). And 10 children with normal occlusion of the same age were added, as a control group. The tongue movements during swallowing, with and without the appliances, were analyzed twice: at the start of, and 6 months after, application of the appliances, by sonography. The results were as follows. 1. The design of a habit-breaker was decided according to the results of a preliminary experiment. The appliance had not only a resin-fence but also a tongue-lifting guide. 2. The upper group showed significant changes in the movement only at the lateral margin of the tongue after 6 months, with and without the appliance. 3. The lower group, however, showed significant changes in the movement at the wider area of the tongue, including the central groove formation, just after applying the appliance, and after 6 months, with and without the appliance. 4. The present results indicated that the modified appliance was more effective in improving tongue function than the conventional habit-breaker. PMID- 11496409 TI - [A new machinability test machine and the machinability of composite resins for core built-up]. AB - A new machinability test machine especially for dental materials was contrived. The purpose of this study was to evaluate the effects of grinding conditions on machinability of core built-up resins using this machine, and to confirm the relationship between machinability and other properties of composite resins. The experimental machinability test machine consisted of a dental air-turbine handpiece, a control weight unit, a driving unit of the stage fixing the test specimen, and so on. The machinability was evaluated as the change in volume after grinding using a diamond point. Five kinds of core built-up resins and human teeth were used in this study. The machinabilities of these composite resins increased with an increasing load during grinding, and decreased with repeated grinding. There was no obvious correlation between the machinability and Vickers' hardness; however, a negative correlation was observed between machinability and scratch width. PMID- 11496411 TI - [The visionary proposals for Japanese radiology in new millennium]. PMID- 11496410 TI - [Hydroxyapatite coating on metal plates using an aerosol jet printing system]. AB - The purpose of the present study was to investigate the properties of the hydroxyapatite (HA) layer formed by an aerosol jet printing system, aimed at application to the field of dentistry. The HA particle was adhered to the metal substrate. The particles of HA were 1 micron or less in size and were packed closely. When the composition of the formed HA layer was compared to that of the raw material powder, oxygen was increased, but Ca was decreased. Moreover, Ca and P were almost evenly distributed in the layer, but oxygen was not. Dynamic hardness of HA coatings was 31.5 +/- 1.1. The peel strength of HA coating was the highest, at 58.7-39.6 MPa, in 10 microns layer thickness. Interface fractures were observed at 10 microns layer thickness, and cohesive fractures were observed for all other layer thicknesses. The wear amount of HA coating by tooth brush abrasion was significantly large at 100 degrees C heating (p < 0.05), compared with at 250 degrees C and 500 degrees C. At any substrate temperatures, the wear amount of stainless steel substrate tended to be smaller than that of titanium substrate. PMID- 11496412 TI - [Clinical applications of functional magnetic resonance imaging]. AB - Despite its immediate success as a tool for basic research, the clinical application of functional MRI(fMRI) is still limited. FMRI has proven useful for presurgical functional mapping of the eloquent cortices. Localization of the sensorimotor cortex by fMRI may be of relatively limited value because the sensorimotor cortex can often be readily localized by means of anatomical methods. In contrast, the language cortices may not be localized anatomically and the language dominant hemisphere has been determined by invasive Wada test. Previous reports have shown that fMRI can be a promising alternative to the Wada test. A recent clinical trial has suggested that fMRI can be used to diagnose Alzheimer's disease in its earliest stage, detecting subclinical deterioration of the memory function. FMRI may be useful to predict the future decline of memory in people with genetic risks. Monitoring of the functional recovery of post stroke brains may be another promising clinical application of fMRI. FMRI has demonstrated functional reorganization of the brain that may be related to the restoration of motor and language functions. PMID- 11496413 TI - [Radiation therapy for painful bone metastases; aiming at optimal treatment schedules]. AB - PURPOSE: To evaluate the pain relief obtained by radiation therapy for painful bone metastases, with a special regard to general condition. MATERIALS AND METHODS: Between June 1998 and May 2000, 54 patients with 86 painful bone metastases were treated with radiation therapy whose effects could be evaluated for a minimum period of 6 months or until death. Treatment schedules were 3 Gy/fraction/day (30-36 Gy/10-12 fractions) in usual cases (61 lesions), 4-8 Gy/fraction/day (8-20 Gy/1-5 fractions) in patients with a poor general condition (9 lesions), and 2 Gy/fraction/day (40-50 Gy/20-25 fractions) in lesions with a large radiation field (16 lesions). RESULTS: Complete pain relief without medication (CR) was achieved in 40 lesions (47%). Significant predictors for CR were primary site (p = 0.0003), performance status (p = 0.0060), pain score (p = 0.0190), narcotic score (p < 0.0001), and prognosis (p < 0.0001), but no difference was found in CR among treatment schedules. No evidence of severe radiation-induced complication was seen. CONCLUSION: General condition (performance status and prognosis) has an influence on pain relief. Compared with the daily 2 Gy protocol, the daily 3 Gy protocol has the advantage of shorter treatment time. The treatment schedule should be assessed in patients with a large radiation field and/or poor general condition. Especially for the patients with poor general condition, combined pain medication should be considered. PMID- 11496414 TI - Evaluation of central nervous system involvement in nasal lymphomas. AB - OBJECTIVES: To clarify the usefulness of evaluating central nervous system (CNS) involvement in patients with nasal lymphomas at the initial staging procedure, and of CNS prophylaxis for patients with clinical stage I/II. PATIENTS AND METHODS: We retrospectively reviewed 43 patients with nasal lymphomas who had been treated from 1973 through 1999. The staging procedure included mainly computed tomography (CT), ultrasonography, gallium scintigraphy, upper gastrointestinal study, magnetic resonance (MR) imaging, and bone marrow biopsy. Forty-two patients received radiotherapy, and 25 patients received chemotherapy. All 38 patients with stage I/II were not subjected to CNS prophylaxis. RESULTS: Four patients demonstrated CNS involvement at the staging procedure. MR imaging demonstrated the tumor had directly infiltrated the skull base in 3 patients, but CT demonstrated CNS infiltration in only one patient. In another patient, cerebrospinal fluid (CSF) cytologic analyses demonstrated CNS involvement, but MR imaging and CT did not. These 4 patients complained of frontonasal pain and/or cerebral nerve dysfunction. No patient with stage I/II developed CNS relapse. CONCLUSIONS: MR imaging and CSF cytologic analyses should be performed at the initial staging of nasal lymphomas, especially in patients with frontonasal pain and/or cerebral nerve dysfunction. Patients with stage I/II might not need CNS prophylaxis. PMID- 11496415 TI - [Assessment of exhaled NO concentration in monitoring radiation pneumonitis in patient who underwent thoracic radiotherapy for lung cancer]. AB - To monitor radiation pneumonitis, we assessed the exhaled nitrogen oxide (NO) level in patient with lung cancer. A 73-year-old man with idiopathic interstitial pneumonitis underwent thoracic radiotherapy without chemotherapy for squamous cell lung cancer (T2N1M0). He showed elevation of exhaled NO level at 30 Gy-50 Gy, after a decrease at 10-20 Gy. He also showed an abnormal shadow on CT examination at 50 Gy. Although exhaled NO may have had the benefit of predicting radiation pneumonitis before severe clinical symptom appeared, he died three months after radiotherapy because of worsening of the radiation pneumonitis. PMID- 11496416 TI - [Effects of the spaceflight factors on the immune system]. AB - The paper summarizes the main trends and milestones of space immunology starting with initial ground-based investigations with animals to recent experiments in piloted and automatic space vehicles data of which have been highlighted in numerous publications. There is a brief description of lines and results of investigations with participation of cosmonauts, and using laboratory animals and cell cultures. The author dwells upon the present-day notion of immune responses to extreme conditions and future topics of investigations. PMID- 11496417 TI - [Medical/psychological problems of pilot's performance in a highly maneuvering flight]. AB - The paper digests phases, principles and content of psychophysiological support to trainees for piloting highly manoeuvrable aircraft. Substantiation is given to the areas of medical/psychological research which are most critical for protection of professional health of flying personnel and safe piloting of the fifth generation aircraft. PMID- 11496418 TI - [Hypokinesia as a risk factor in extreme conditions]. AB - The paper describes the specific clinical/physiological symptoms of hypokinesia developing in bedrested female and male subjects. It was shown that hypokinesia calls forth a peculiar syndrome that substantially impairs human performance. Experimental and clinical data indicate that hypokinesia should be regarded as a serious risk factor stimulating pathological alterations in the visceral organs. Proceeding from this assumption, investigations along the line should be persevered in order to contribute to the general theory of key reactions of organism to hypokinesia, and search for preventive means and methods to be applied both to space medicine and public health. PMID- 11496419 TI - [Microgravity and development of the mammals: problems results prospects]. AB - The author reviews results of 20-yr in-space and laboratory investigations into the effects of hypergravity (2 g) and hypokinesia (immobilization, suspension, water deprivation). It was found out that the compensatory and adaptive potentials of mammalians (Wistar rats) in microgravity are sufficient to let proceed pregnancy, and activate anabolic processes associated with fetus growth and water-salt homeostasis maintenance. In a number of aspects effects of microgravity on the mother-fetus systems appear to be less significant than of various factors on Earth including the conventional models of microgravity. In all types of experiments, fetus state was determined by mother's well-being, compensatory and adaptive potentials. Under the disadvantageous conditions, survival of fetus cost dramatic shifts in mother's organism. Outlined is a long range program of research on the problem and feasibility of an experiment in which mammalians will pass the overall life cycle in space flight. PMID- 11496420 TI - [The evaluation system and forecasting of the cosmonaut's health: its prospects for development]. AB - Principles and modalities that had been adopted for health evaluation of the first space crews set a basis for the system of medical monitoring of orbital missions longer than a month. Long-term shifts in space called forth the development of as diagnostic so prognostic approaches to health evaluation. One of the methodological fundamentals was the use of the blood circulation system as an indicator of adaptive reactions of the whole body. Data about the cardiovascular reactions to extended exposure to the spaceflight factors on space complex Mir are of major importance for improvement of the effectiveness of medical operations aboard the International space station. Some of the methods applied on Mir can be perfected to meet medical requirements to a mission to Mars. PMID- 11496421 TI - [Tolerance of +Gx by MIR 22 -- 27 main crew in space flights]. AB - Data on +Gx tolerance by 16 cosmonauts-members of the Mir 22-27 main crews in missions from 8 up to 380 days in duration are reported. All cosmonauts showed good tolerance of g-loads during insertion into orbit. Owing to the use of anti-g suit Centaur and water-salt supplements prior to deorbiting, tolerance of +Gx in the course of descent was satisfactory in the majority of the cosmonauts and low only in one person. As compared with short missions (8 to 21 days), strain of the physiological systems exposed to the deorbit +Gx loads following an extended period in microgravity (186 to 380 days) was greater and manifested by more often arrhythmias, hard breathing and speech, vestibulo-autonomous reactions, petechial hematomas into the back tegmentum, and more severe sinus tachycardia. As a rule, there were no symptoms indicative of poor g-tolerance (visual disorders etc.) which speaks for effectiveness of the anti-g measures in most of the cosmonauts. However, prognostically bad cardiac rhythm disturbances (multitude of single, group and systemic polytope ventricular extrasystoles) registered in one cosmonaut on the deorbit phase mean that efforts should be redoubled to improve countermeasures against the deconditioning of organism by microgravity, particularly the cardiac rhythm disturbances. PMID- 11496422 TI - [From 108 minutes to 438 days and further on... (on the 40th anniversary of Iu. A. Gagarin's flight)]. AB - The progress of science and technology at the end of the XIXth and first half of the XXth century paved the way to start space exploration by humanity. The flight of Yu. A. Gagarin on April 12, 1961 was one of the history watersheds that had a great many of social implications. Piloted missions to space demonstrated the possibility for humans to adapt to the spaceflight factors which, nonetheless, can provoke various unfavorable reactions, particularly on return to Earth. Step by-step extension of mission length paralleled enhancement of the methods of monitoring of crew health, life support systems, and development of modalities to maintain crew health and performance. The paper contains brief discussion of data acquired in short- and long-term missions including the 438-d mission of cosmonaut-physician V.V. Polyakov. The final section is devoted to the mainstream problems of future piloted space programs. PMID- 11496423 TI - [Prevention of altitude decompression sickness during simulation of egress from the cabin of space vehicle with hypobaric artificial atmosphere]. AB - Based on the proposed semiempiric nomograms, the study was aimed at selection and investigation of normoxic N2-O2 hypobaric atmosphere (HA) at 93-73 kPa, and evaluation of effectiveness of the gas mixture against the altitude decompression sickness (ADS) during 6-hr simulation of extravehicular activities at 37 kPa without prebreathing. Subjects were 22 healthy males from 20 to 50 yr. old. In all, 115 experiments were performed. Twenty-four hours in HA at 93 kPa and subsequent transition to 37 kPa led up to development of ADS symptoms by one subject in one of 27 experiments (3.7%), and emergence of venous gas bubbles (USI) with the intensity of 2, 70, and 19 points by the Spenser scale in 9 experiments (33.3%). Lower HA pressures (86 kPa, 80 kPa and 73 kPa) did not give rise to ADS though the occurrence of venous GB distinctly tended downward (30.6%, 14.3% and 11.8%) and so did the intensity of their entry into the pulmonary circulation (2, 40, 18 points; 1, 80, 19 points, and 2, 00, 15 points, respectively). Minimal ADS risk was observed in HA at 80 kPa and 73 kPa and did not reach 4% at the confidence level of 95%. PMID- 11496424 TI - [Psychological aspects of training for and implementation of a piloted expedition to Mars]. AB - Reviewed are the modern notions of the main psychological concerns regarding a mission to Mars which are maintenance of mental health, high professional reliability, and successful readaptation of the Martian crew on return to Earth. The unprecedent trail-blazing interplanetary mission distinguished by autonomy, long period, international crew and a broad range of tasks to tend will set particularly difficult requirements to the psychological support system. Among them prioritized are purposeful character building and training of future crew members already in childhood and youth. The primary goal is to bring up personality with anthropocosmic thinking as a professional quality that will eventually determine personal and professional reliability. Within the system of psychological support, the central place should be given to monitoring of crew mental heath and behavior, and preventive an corrective measures. The longer Martian day (24 hr. 36 min.) rises the issue of effective adaptation to this factor. Special simulators will be required aboard the vehicle to help the crew refresh skills in manipulation of the dynamic board systems. The program of psychological support to the Martian crew must be thoroughly tested in model studies (Antarctic field, ground-based experiments, orbital flights, Lunar base). PMID- 11496425 TI - [Radiation risk in aircraft altitude flights]. AB - The radiation risk in aircraft altitude flights was evaluated from calculated dose fields at the altitude of 10-30 km 0-90 degrees North modulated by the galactic cosmic radiation (GCR) in quiescent heliophysical periods. Results of dose measurements in flight are discussed in the light of present-day radiation limits. The authors suggest that pilots should be subsumed under the category of professionals who work with sources of ionizing radiation. PMID- 11496426 TI - [Flight personnel rehabilitation]. AB - Presented are results of the body functional evaluation on a background of the most common syndrome of neurocirculatory dystonia and spinal osteochondrosis in flying personnel. Execution of tone-strength stimulating physical exercises for 10 to 12 days noticeably improved professional health. PMID- 11496427 TI - [The first cosmonaut of Earth (personal memories)]. PMID- 11496428 TI - [The history of the development of medical approaches to the selection of the first cohort of space flight candidates]. PMID- 11496429 TI - [The 28th public and scientific readings in memory of Iu. A. Gagarin]. PMID- 11496430 TI - [Significance of retardation of abnormal uptake of iodine-123-beta-methyl-p iodophenyl-pentadecanoic acid myocardial scintigraphy in patients with vasospastic angina]. AB - OBJECTIVES: This study investigated retardation of abnormal uptake of iodine-123 beta-methyl-p-iodophenyl-pentadecanoic acid(BMIPP) scintigraphy in patients with vasospastic angina. METHODS: Twenty-three patients with vasospastic angina showed abnormal uptake of BMIPP before medical treatment and had coronary vasospasm induced by acetylcholine. The patients were divided into two groups according to uptake of BMIPP after medical treatment: retardation of abnormal uptake of BMIPP(Group R, n = 4) and normal uptake of BMIPP(Group N, n = 19). Frequency of chest pain, medical treatment and autonomic nervous activity were compared between the two groups. Furthermore, the frequency of chest pain and uptake of BMIPP in group R were obtained after intensive medical treatment. Autonomic nervous activity was evaluated by heart rate variability on Holter electrocardiography. Heart rate variability contained high-frequency elements(HF; 0.15-0.4 Hz) and low-frequency elements(LF; 0.04-0.15 Hz). LF/HF was estimated for sympathetic nervous activity and HF was estimated for parasympathetic nervous activity. Daytime and nighttime autonomic nervous activity were compared between the two groups. RESULTS: The frequency of chest pain was higher in Group R than in Group N(p < 0.05). Medical treatment was not different between the two groups. Circadian variation of sympathetic and parasympathetic nervous activity were absent in Group R. During the nighttime, Group R showed higher sympathetic nervous activity(p < 0.05) and lower parasympathetic nervous activity(p < 0.01) than Group N. The frequency of chest pain was significantly lower after intensive medical treatment(p < 0.05), and uptake of BMIPP returned to normal in Group R. We suspected that the disorder in autonomic nervous activity was more severe in Group R, and thus induced coronary vasospasm. CONCLUSIONS: Retardation of abnormal uptake of BMIPP in patients with vasospastic angina indicates poor control of coronary vasospasm. Uptake of BMIPP is useful in the evaluation of coronary vasospasm control in such patients. PMID- 11496431 TI - [Prevalence of cervical and cerebral atherosclerosis and silent brain infarction in patients with multivessel coronary artery disease]. AB - OBJECTIVES: This clinical study investigated the prevalence of cervical and cerebral atherosclerosis and silent brain infarction in patients with coronary artery disease. METHODS: Cervical and cerebral magnetic resonance angiography(MRA) was performed in 133 patients (98 males, 35 females, mean age 65.3 years) with suspected coronary artery disease, who were divided into a zero- and one-vessel disease group(n = 71) and a two- and three-vessel disease group(n = 62) depending on the number of major coronary branches with 75% or more stenosis. The MRA lesion was defined as more than 50% stenosis. Magnetic resonance imaging(MRI) of the brain was performed within 1 week of MRA in 78 patients without symptomatic stroke and atrial fibrillation. Silent brain infarction on MRI was defined as a focal high intensity area on T2-weighted images larger than 3 mm. RESULTS: The prevalence of MRA lesions was significantly greater in the two- and three-vessel group than in the zero- and one-vessel group(53% vs 14%, p < 0.01). The prevalence of MRI lesion was significantly higher in the two- and three-vessel group than in the zero- and one-vessel group(77% vs 36%, p < 0.01). The size and number of the MRI lesions were also significantly greater in the two- and three-vessel group than in the zero- and one-vessel group(p < 0.01). Neither age nor percentage of male gender was different between the groups. Diabetes mellitus was the common risk factor for coronary artery disease, MRA lesion and MRI lesion. CONCLUSIONS: Cervical and cerebral atherosclerosis and silent brain infarction are frequently observed in patients with multivessel coronary artery disease. PMID- 11496432 TI - [Relationship between endothelial dysfunction and nitric oxide production in young male smokers]. AB - OBJECTIVES: Endothelial function in the brachial arteries is impaired in smokers. However, little is known about this condition in young adult men. The relationship between nitric oxide(NO) production and the endothelial function was investigated in young smokers and compared with non-smokers. METHODS: Flow mediated vasodilation of the brachial artery during reactive hyperemia was examined in 10 young smokers(mean age 31 years) and 12 control subjects(mean age 28 years). The vasodilator response in the brachial artery was measured by ultrasonography, and blood samples were obtained from the right cephalic vein. Blood samples were taken at baseline, 30 sec after cuff deflation, and before and 5 min after 0.3 mg of nitroglycerin administration. Blood flow was calculated by multiplying mean flow velocity and vessel cross-sectional area. Plasma NOx(nitrate + nitrite) levels were measured, and the percentage change of NOx production(delta NOx) was calculated as follows: delta NOx(%) = [(NOx concentration at peak flow-mediated vasodilation or after 0.3 mg nitroglycerin administration) - baseline NOx concentration)] x 100/baseline NOx concentration. RESULTS: Percentage changes in diameter of the brachial artery, NOx production and delta NOx in response to nitroglycerin were not statistically different between the two groups(smokers: 27.6 +/- 8.0 mumol/l, control subjects: 34.0 +/- 8.7 mumol/l). However, percentage change of flow-mediated vasodilation during reactive hyperemia in the young smokers was significantly smaller than that in the control subjects(4.8 +/- 2.7%, 9.1 +/- 5.3%, respectively, p < 0.05). Moreover, delta NOx during reactive hyperemia in the smokers was significantly smaller than that in the control subjects(388.8 +/- 90.2%, 738.0 +/- 284.5%, respectively, p < 0.05). CONCLUSIONS: The impaired response to reactive hyperemia in young smokers might be associated with decreases in flow-dependent NO production. PMID- 11496433 TI - [Effect of percutaneous transvenous mitral commissurotomy for the preservation of sinus rhythm in patients with mitral stenosis]. AB - OBJECTIVES: Atrial fibrillation is frequently associated with mitral stenosis and is considered to be an unfavorable factor for the long-term prognosis. The efficacy of percutaneous transvenous mitral commissurotomy(PTMC) was examined for the preservation of sinus rhythm in patients with mitral stenosis after PTMC. METHODS: Long-term clinical data after PTMC were obtained from 71 patients who had undergone PTMC from March 1989 to September 1999. Eighteen patients in sinus rhythm before PTMC were divided into two groups: the SR group(n = 5) who remained in sinus rhythm, and the Af group(n = 13) who showed change from sinus rhythm to persistent or paroxysmal atrial fibrillation after PTMC. RESULTS: Age, sex, mitral valve area(1.4 +/- 0.3 vs 1.2 +/- 0.3 cm2), mean mitral pressure gradient(14.3 +/- 5.5 vs 12.6 +/- 5.9 mmHg), mean left atrial pressure(15.9 +/- 7.6 vs 19.0 +/- 7.7 mmHg), left ventricular end-diastolic pressure(7.5 +/- 2.8 vs 9.3 +/- 3.9 mmHg), left ventricular end-diastolic volume index(77 +/- 13 vs 82 +/ 14 ml/m2), left ventricular ejection fraction(60 +/- 6% vs 55 +/- 4%) and cardiac output(5.1 +/- 0.4 vs 4.9 +/- 0.8 l/m2) before PTMC were not different between the two groups. Changes in mean mitral pressure gradient, mean left atrial pressure and cardiac output immediately after PTMC were not different statistically. Mitral valve area immediately after PTMC was significantly greater in the SR group compared to the Af group(2.3 +/- 0.3 vs 1.8 +/- 0.3 cm2, p < 0.05). The change in mitral valve area was also greater in the SR group(1.0 +/- 0.2 vs 0.6 +/- 0.4 cm2, p < 0.05), but there was no statistical difference in the percentage change of mitral valve area between before and immediately after PTMC(SR group 78 +/- 35% vs Af group 50 +/- 35%). End-diastolic pressure, end diastolic volume index and ejection fraction immediately after PTMC were not statistically different. CONCLUSIONS: The final mitral valve area immediately after PTMC in the patients with mitral stenosis in sinus rhythm, but not the changes of mean mitral pressure gradient, mean left atrial pressure or cardiac output, is important for the maintenance of sinus rhythm. PMID- 11496434 TI - [Female carrier of Duchenne muscular dystrophy presenting with secondary dilated cardiomyopathy: a case report]. AB - A 48-year-old female carrier of Duchenne muscular dystrophy had developed congestive heart failure but had no skeletal muscle symptoms. She was admitted to our hospital complaining of palpitation in December 1998. Her three sons had Duchenne muscular dystrophy. Neurological examination was unremarkable with no evidence of muscle weakness. Serum creatine kinase level was slightly increased. Echocardiography showed severe left ventricular dysfunction. Coronary angiography showed no abnormalities. Left ventriculography showed generalized hypokinesis and left ventricular ejection fraction was 28%. Dystrophin immunostaining of the skeletal muscle biopsy specimen showed a mosaic pattern. The dystrophin negative fibers were scattered among positive fibers. Cardiomyopathy is the only clinical manifestation of dystrophin gene mutation in carriers. Beta-blocker therapy(carvedilol 5 mg/day) was effective in this patient. PMID- 11496435 TI - [Apical ballooning by transient left ventricular dysfunction (so-called "ampulla" cardiomyopathy) associated with therapy for acute pulmonary thromboembolism: a case report]. AB - An 86-year-old-woman presented with apical ballooning left ventricular dysfunction associated with therapy for acute pulmonary thromboembolism. She was referred to our hospital for advanced treatment for her shock state due to acute pulmonary embolism with normal left ventricular(LV) function. Her condition was stabilized using a percutaneous cardiopulmonary support system. Suction embolectomy was successfully carried out after pulmonary arteriography. After the therapy, echocardiography revealed apical ballooning and hyperkinesis of the base(LV ejection fraction = 28%), although coronary arteriography showed no fixed stenosis. LV wall motion significantly improved on day 3(LV ejection fraction = 45%). Pulmonary embolism relapsed on day 5 in spite of anticoagulation treatment. She died of multiple organ failure on day 9. Autopsy findings indicated no sign of myocardial infarction or myocarditis, patchy appearance of myocardial contraction band necrosis and few migrated lymphocytes. The mechanism for the flow mis-matched LV dysfunction remains unknown. The probable explanations include non-ischemic stress such as catecholamine or neurogenic stress, and possibly ischemic stress or ischemia/reperfusion injury. PMID- 11496436 TI - [A 73-year-old man with intermittent claudication after stenting in the external iliac artery]. PMID- 11496437 TI - Putting retinal exams for diabetics in the primary care arena. AB - Only about half of all diabetics get their recommended annual eye exams. Until now, little progress has been made in boosting the number of diabetics who are compliant in getting this annual exam done. However, a new device that enables the exams to be completed in the PCP's office is getting rave reviews from early users. PMID- 11496438 TI - Solid outcomes show e-health and chronically ill senior populations are compatible. AB - Why? Because at least one new study suggests that many seniors not only embrace Internet-based models, but in fact prefer them over more traditional DM approaches. Further, Internet-based programs often come with additional benefits that cannot be delivered via interactive voice response (IVR) or other delivery models. There are challenges involved with an e-health approach to be sure, but some providers are finding that they are rewarded for their efforts with solid outcomes. PMID- 11496439 TI - Researchers hot on the trail of modest interventions to boost depression care. AB - There is no question that providers need to do a better job of diagnosing depression in primary care settings. However, research is also now focused on uncovering better management strategies for newly diagnosed patients and those patients at highest risk for relapse. A flurry of work is underway to test modest follow-up strategies that can boost outcomes and improve quality of life. Early results suggest the approach is on target. PMID- 11496440 TI - Competing health plans collaborate to boost asthma care. PMID- 11496441 TI - Are you prepared for terrorist attack? New report tells how to train ED staff. AB - A new report from the American College of Emergency Physicians and the Department of Health and Human Services gives specific training objectives for ED staff to prepare for casualties of nuclear, biological, or chemical (NBC) incidents. Courses tailored to the objectives will be available in approximately one year. You should include decontamination scenarios in disaster drills. Be certain that NBC incidents are addressed in your hospital's disaster plan. Provide adequate training for staff with an instructor who has been trained by the Department of Defense or has undergone training in hazardous materials. PMID- 11496442 TI - How to handle abusive colleagues. PMID- 11496443 TI - Follow up with patients by telephone. AB - Giving patients a follow-up telephone call can improve satisfaction, improve outcomes, and even save lives. Patients who left the ED without being seen can be encouraged to return if the problem has persisted or worsened. The nurse who makes the follow-up calls should be trained in giving advice over the telephone and be a good listener. Patients react positively to receiving follow-up calls. PMID- 11496444 TI - ["Integrating psychiatry, psychoanalysis, neuroscience." A new tone at the annual meeting of the American Psychosomatic Society and American Psychiatric Association]. PMID- 11496445 TI - [A consecutive study of patients with irritable bowel disease in two tertiary referral centers]. AB - OBJECTIVES: We investigated the nature of illness behavior and the meaning of emotional deficiencies during childhood in patients with irritable bowel diseases (IBS). DATA: A consecutive study in two tertiary referral centers was conducted with 48 patients suffering from irritable bowel diseases (IBS) and 91 patients with inflammatory bowel disease (IBD). METHODS: The diagnosis of IBS was made by following the Manning criteria, a positive diagnosis of IBD was established through physical, endoscopic and radiologic examinations and was confirmed histologically. Psychological data were obtained by structured psychiatric interviews and psychological self-report measures (GBB). RESULTS: We found that the rate of physician visits given in the course of the disease, is increased for those having irritable bowel disease (IBS). These patients are dissatisfied with the physicians and prone to psychophysiological complaint. In the daily routine and in occupation they are more impaired than those with inflammatory disease. This finding emphasizes in particular that patients with irritable bowel diseases (IBS) have experienced emotional deficiencies in childhood as an after effect of loss, divorce of the parents etc. CONCLUSIONS: Implications for doctor-patient relationship and the necessity for epidemiological studies in Germany are discussed. PMID- 11496446 TI - [Effects of a brief couples therapy intervention in coronary rehabilitation. A controlled study]. AB - The effects of a brief couple therapy intervention in cardiac rehabilitation was investigated in a randomised controlled trial. The treating physician and a psychologist conducted an intervention of two to four sessions with 19 patients with coronary diseases at the IIa stage according to WHO and their partners. The 21 patients in the control group received standard care with no additional intervention. Depressive symptoms, global self-rating of health status and disease related concerns were assessed at the beginning of training and nine months later. Patients in the experimental group more often stated to feel better immediately after the intervention. Over the observation period they showed significantly more favour-able changes in observer rated depressive symptoms, self-rated health status and concerns. Patients and partners in the experimental group assessed the outcome of rehabilitation significantly more positive after nine months. The findings suggest that a very short couple therapy intervention, which can be administered in routine care, can have positive effects in cardiac out-patient rehabilitation. Larger studies are required for testing whether the effects of such interventions are restricted to depressive symptoms and subjective appraisals, or whether other outcome criteria such as mortality can also be influenced. PMID- 11496447 TI - [Coping in patients with malignant glioma in the course of radiation therapy]. AB - Twenty-one patients with malignant glioma were interviewed in the course of radiation therapy (before start, in the middle and at the end of treatment and six weeks later). We used the "Freiburger Fragebogen zur Krankheitsverarbeitung (FKV)", an assessment of coping strategies, the "State-Trait-Anxiety-Inventory (STAI)", the "Beck-Depression-Scale (BDI)", and the QLQ-C 30 questionnaire of the EORTC. RESULTS: The coping strategies of our patients are comparable with other cancer patients. They are mainly characterized by "self-encouragement", "compliance" and "trust in the treating physician". Anxiety was low and showed no significant changes. Depressivity was higher than in the normal population, however, it also showed no significant changes in the course of therapy. Quality of life scores remained constant, despite an increase of fatigue. In our patients with malignant glioma, the influence of radiation on coping, anxiety, depression and quality of life seems insignificant in comparison to that of the diagnosis of cancer. PMID- 11496448 TI - [Publication practices of academics in medical psychology and in psychosomatics and psychotherapy]. AB - When qualifying for higher academic positions junior academics face increasing demands for submitting papers for publication. The criteria for assessing these publications are presently under discussion. Contributions to international English language journals are more highly regarded, and it has become indispensable to have papers published in journals listed in SCI and SSCI. The question remains whether these criteria are valid to judge academic qualifications. Whereas one criterion for validity may be the publication practice of the present academic representatives, it appears that to some extent the chairs themselves would not fulfill the requirements for academic qualification today. Results regarding this are presented and discussed. PMID- 11496449 TI - [Psychosomatic research in intensive care medicine. An overview]. AB - After a brief historical introduction the literature that has appeared on psychosomatic research in intensive care units since the middle fifties is reviewed. The main focus in published studies is directed towards subjects like the patients and their specific diseases as fatal illness or coronary heart disease as well as family needs and interpersonal assessment of staff members. Results show the importance of helpful and considerate way of working among staff members to reduce stress. Supervision is recommended and supportive psychotherapy for patients with severe emotional distress are suggested. PMID- 11496450 TI - [Central relationship patterns in comparison with different objects]. AB - In the present study the Relationship Episode Paradigm Interviews of 70 female patients with different psychoneurotic diseases were analysed with respect to object-specific patterns with the CCRT method. The most frequent categories are the same in all relationship episodes and in subsamples of relationship episodes with mother and father. These categories are also predominant in episodes with women and men. Relationship episodes with mother do not differ from episodes with father, and relationship episodes with women are not different from episodes with men. But there are substantial differences in relationship episodes with the mother and women and between episodes with the father and men. Patients recount much more positive relationship patterns with women and men than with their parents. This could be understood as a hint of interpersonal resources. PMID- 11496451 TI - [Speidel, Hubert: Writing process, PPmP and I in the middle of the word (Editorial, issue 12).PPmP Psychother Psychosom med Psychol 50: 445 (2000)]. PMID- 11496452 TI - Private health care in Australia: why the government should intervene. PMID- 11496453 TI - Public health advocacy--determining a role for staff of a public hospital. AB - This study explored the public health advocacy role of staff employed at a major South Australian specialist public hospital that had made an organisational commitment to health promotion. It was concluded that staff did have a role to play in public health advocacy, though the issues pursued primarily related to disease and injury prevention rather than the broader social determinants of health. Staff gained valuable experience in the political processes of public health advocacy. The hospital had to develop organisational infrastructures to support staff so they could undertake public health advocacy on behalf of the hospital. It was also necessary for the hospital to implement other changes in order to 'get its own house in order' before issues could be addressed in the broader community. PMID- 11496454 TI - Emergency and primary care at a Melbourne hospital: reasons for attendance and satisfaction. AB - The reasons for attendance, presenting health problems, functional status, pain and severity, and satisfaction with emergency and primary care were examined using routinely collected data and an interviewer-assisted survey of patients. Patients attended, mostly after hours, because they believed their health problems required hospital-based management. GPs referred for admission and further evaluation. Ethnicity, employment status, gender and age contributed to differences in access, morbidity and pain scores. Pain scores, functional status and English language skills influenced satisfaction. Culturally sensitive hospital- and community-based clinicians are important to promote better services, after-hours care, referral and triage. It is essential to have appropriate policy and legislation, adequate infrastructure and resources, good communication strategies, telecommunication technology, explicit evidence-based protocols for shared care, referral and triage and ongoing training and support for clinicians and consumers. PMID- 11496455 TI - Treatment for childhood acute lymphoblastic leukaemia: the fathers' perspective. AB - Research on parental adaptation to a child's chronic illness is still scant, and this is particularly so in relation to the experience of treatment for paediatric Acute Lymphoblastic Leukaemia (ALL). The work that does exist on parental reactions tends to conflate maternal responses with paternal responses, as fathers are usually seen as having a secondary role. Consequently, little is known about how fathers cope with treatment for childhood ALL. The present discussion seeks to make a contribution to this area by presenting findings on the paternal experience of treatment for paediatric ALL from a longitudinal study conducted at Royal Children's Hospital in Brisbane, Queensland. The findings from this research clearly indicate the emotional pain that fathers face in their struggle to accept the diagnosis of a serious, life-threatening illness such as ALL in their child. The findings challenge the notion of the make stereotype by showing that the shock of diagnosis, the emotional pain of coping with the illness, the expression of pain through tears, the desire to be with the child, the struggle to cope with the medical interventions, and concerns about other family members are not gender specific, but are rather issues common to both parents. PMID- 11496456 TI - From efficacy to effectiveness: managing organisational change to improve health services for young people with deliberate self harm behaviour. AB - Repeat Deliberate Self Harm is a recognised risk factor for completed suicide and therefore reduction by effective health service response represents a valid contribution to suicide prevention. However, only a small fraction of people with deliberate self harm presentations to general health settings actually reach specialist mental health follow-up appointments. Therefore, even if responses at that point are known to be effective they do not make a significant contribution to reducing repeat self-harm overall. We describe health system organisational change strategies to improve health service engagement for the target group, and present data demonstrating the effectiveness of these strategies. PMID- 11496457 TI - And on the cover of the guide were two words in red--don't panic. PMID- 11496458 TI - Mongolia's system-wide health reforms: lessons for other developing countries. AB - Mongolia is a poor country that lost 30% of its GDP when the Soviet Bloc collapsed in 1990. Its health care system had the typical weaknesses of centrally planned economies--quantity rather than quality, excessive medical specialisation, dominance of the hospital sector, weak policy and management capabilities, little community participation in decision making, and so on. This paper describes Mongolia's attempts to resolve these problems through a radical program of reform that began in 1998. There have been significant successes in spite of almost overwhelming difficulties, and this may be a consequence of the strong sense of community that has been present for five hundred years and re emerged intact at the end of 70 years of Soviet dominance. We argue, however, that good design and skillful implementation of the reform program may have made a contribution. Its notable features have included the use of a comprehensive and integrated model rather than piecemeal reform, the generation of political support for change through social marketing campaigns, a team approach using local and international experts, and co-ordination of international donor activities. Some of these features may be relevant to other transitional and developing countries. PMID- 11496459 TI - The Illawarra Coordinated Care Trial: better outcomes with existing resources? AB - The Illawarra Coordinated Care Trial was one of nine Australian trials undertaken to see whether different models of coordinated care could improve the health of people with multiple service needs within existing resources. This paper summarises the findings of an extensive local evaluation and discusses the impact of the trial on clients and service providers. It examines the main findings related to the principal trial hypothesis and points to lessons that might inform the next round of trials. PMID- 11496460 TI - The changing attitudes of health professionals and consumers towards a coordinated care trial--SA HealthPlus. AB - The national coordinated care trials have been a vehicle for health reform in Australia, driven by escalating health care costs and projections of an ageing population. The first round of trials conducted between 1997 and 1999 set the trials a challenge to reduce financial and system barriers to enable health professionals in all sectors and consumers to develop service delivery models which would give better outcomes for patients within existing resources. As part of a change management strategy, the developers of the SA HealthPlus trial assessed the attitudes of health professionals and consumers involved in designing the projects which made up the larger trial, prior to trial development and twelve months later. This paper reports on the results of the survey and how initial enthusiasm gave way to appropriate anxiety as the complexities of creating a new system of care from reactive to prospective patient centred care planning, became a reality. The survey enabled trial developers to show evidence of acceptability for the new model of care and identify areas of concern and appropriate strategies for the project teams. This type of survey and the issues identified may be of benefit to the second round coordinated care trials and health regions aiming to initiate coordinated care programs. PMID- 11496461 TI - To HITH or not to HITH: making a decision about establishing hospital in the home. AB - Hospital in the home is increasingly being considered as an alternative for the provision of acute care. This article provides an overview of Hospital in the Home in Australia, discussing some of the issues that should be considered when determining whether to establish or fund hospital in the home programs such as whether efficiency is increased, care is improved and whether patients perceive more choice. These issues are discussed in the context of a transparent funding strategy that is aimed at achieving predefined goals and objectives. PMID- 11496462 TI - Macarthur model for ambulatory services. AB - The Macarthur Health Sector has embraced the concept of Ambulatory Care within all clinical streams. The Macarthur Model for Ambulatory Services is multi faceted and has sought to encompass and combine the best features of many established services throughout Australia and the world. The results is one integrated, cohesive model that allows patients multiple points of entry and exit. Ambulatory Care has developed an essential link in the continuum of care from acute 'illness' hospital-based to maintenance community 'health' care. The philosophy of shared responsibility between patient, carer, general practitioner and a hospital specialist team underpins the model developing in Macarthur. PMID- 11496463 TI - Health needs assessment and the ecology of care: a research note. AB - A Health Needs Assessment project was undertaken in 1999 for The Royal New Zealand Plunket Society (Inc.), a voluntary community organisation that provides child and family health promotion to 50,000 new babies each year nationwide. Specially qualified registered nurses, community workers and volunteers deliver the service that includes home visiting as part of universal health care. The aims were to verify and refine criteria for health needs assessment by nurses for the ongoing development of practice. Methods used enabled participatory action by child and family health nurses (Plunket nurses). The context was health promotion for apparently well children and their families as part of the Ministry of Health universal Well-Child Tamariki Ora program. The results included identification of a series of health needs assessment criteria, and direction to further develop practice. As a result of the project, it was possible to incorporate the four major concepts in the 2000 revision of the Plunket Health Record. The challenge for child and family health promotion in New Zealand is to assess health need at a family level with the focus on improving child health outcomes. Nurses who use standard data collection items to assess health needs are also likely to identify subsequent interventions more explicitly. Therefore the main benefit of the project is the potential to improve clinical practice. PMID- 11496464 TI - Coping with health care reform. AB - This paper describes the evaluation of a case study of health service reform in southern Adelaide. A mail survey, interviews and focus groups were used to gain an understanding of health care providers' perceptions and experiences of reform. The paper discusses lessons learnt about health service reform and its impact on people working in the health system. It finds that continual change that is not based on a well-articulated vision is likely to lead to 'reform fatigue' and low morale. An action research approach can be used by researchers to help managers and staff understanding the context in which reform is happening, and thus give support to organisational learning. PMID- 11496465 TI - Recognising the changing boundaries of illness in defining terms of chronic illness: a prelude to understanding the changing needs of people with chronic illness. AB - Definitions of chronic illness do not reflect the changing nature of chronic illnesses. When definitions in the literature, which guide and inform thinking in a field, remain static they are in danger of creating stereotypes. This can have an adverse influence on the care of people with chronic illness. Debates over the use of terms associated with chronic illness will lead to a better understanding of the place of chronic illness in the world of health and illness and ultimately lead to services that better meet the needs of consumers. PMID- 11496466 TI - Funding arrangements for pharmaceuticals: can economic evaluation promote efficiency? PMID- 11496467 TI - The pharmaceutical situation in the Philippines. PMID- 11496468 TI - Evidence-based priority setting. AB - This paper describes evidence-based priority setting and resource allocation undertaken by a Division of the Women's & Children's Hospital, Adelaide during 1998-1999. We describe the methods used to combine program budgeting marginal analysis (PBMA), evidence based and "community values" approaches into one decision-making framework. Previous organisational changes involving the formation of multidisciplinary team and program management were pivotal in setting a framework to successfully complete the priority setting process. PMID- 11496469 TI - Are Australia's drugs too cheap for our own good? PMID- 11496470 TI - Exploring distributive justice in health care. AB - The allocation of resources to providers and the way in which the resources are then prioritised to specific service areas and patients remain the critical ethical decisions which determine the type of health system a community receives. Health care providers will never be given enough resources to satisfy all the demands placed upon them by a community that is becoming increasingly informed and demanding. This paper discusses the matter of justice as it relates to the distribution of health resources. It translates the theoretical constructs of distribution into a practical situation that arose at The Geelong Hospital. It is important to emphasise that the aim of giving the example is not necessarily to provide the right answer but rather to assist in determining what ought to be the questions. PMID- 11496471 TI - Organisational change: a methodology to uncover the business idea. AB - A study was undertaken to identify the "Business Idea", as defined by van der Heijden (1996), in The Family Planning Association of WA Inc (FPWA) which is a Non-Government Organisation (NGO) in Perth, Western Australia. This organisation was chosen as, along with many other NGOs, it was undergoing major changes in its funding, role and required outcomes. A qualitative interpretivist single case study methodology employing grounded theory research principles and methods was used to study the Business Idea framework in this setting. Thirty-four members of FPWA's staff were interviewed and data was managed using NUD*IST4 and Decision Explorer data storage, data retrieval and graphical reproduction facilities. Results indicated that images of the Business Idea model within FPWA were largely consistent across all staff levels excepting members of the Board of Management. Changes within the organisation were impacting heavily on staff, who needed to be assisted over the transitional phase. Strong leadership and corporate direction were identified as essential if the FPWA was to balance the strongly held sense of social justice amongst its staff with a need for greater productivity efficiency and accountability across the organisation. PMID- 11496472 TI - Increasing the efficiency and flexibility of capital funding for public and private hospitals. AB - This paper outlines the recent history of capital funding in the health sector in Australia. It focuses on the trends in capital expenditures and the roles of the public and private sectors. The demand for future capital funding will depend upon a number of factors, including the state of the current capital stock and uncertain future impacts of technology and changes in patient demands. Because of these uncertainties, capital funding for the future must be flexible to meet any changes in operations and demand. The paper highlights the possible use of joint ventures between various public and private organisations, based on the principles of competition and cooperation. PMID- 11496473 TI - Factors influencing patients' length of stay. AB - This study was conducted to evaluate the ability of AN-DRG version 3.1 to predict variation in patients' length of stay in hospital (LOS) and identify other factors that can influence the LOS by using routinely collected hospital morbidity data. A total of 18 DRGs that comprised 4,589 episodes were analysed. Multiple regression was used to model length of stay as a function of a number of independent variables. Overall only 37.6% of variation in mean length of stay could be explained. DRGs predicted 30% of the total variation. Other factors such as age, payment classification, source of referral, specialty of doctor, and ethnic group also influenced patient length of stay. It was concluded that the limited explanation was a consequence of a lack of a better indicator of severity within DRGs. PMID- 11496474 TI - Australia's pharmaceutical benefits system: flawed but improving, and better than anywhere else. PMID- 11496475 TI - Re-engineering clinical records for production control. 1917. PMID- 11496476 TI - Seeding information management capacity to support operational management in hospitals. AB - There are vast amounts of regularly reported data in the information systems of hospitals, state and federal governments. The increase in accessibility offered by platforms such as the Health Information Exchange (HIE) in New South Wales (NSW) creates a new level of opportunity. Administrative data can also speak to clinical and managerial issues. The capacity to mine these data and use the information for improving quality and efficiency has not been well developed at the "coal face" of operational management. Whilst it has been both possible and useful to track utilisation of services to hospitals and patients as cost and volume, it has not been of interest to track these same data to the operational locus of care--the nursing unit, the operating room, the imaging department. With HIE-type systems, the information is now more readily available and operational managers know this. The challenge is to develop the interdisciplinary capacity to query administrative data to facilitate clinical and managerial decision-making. We report here a possible model of a systematic approach to developing this capacity and some of the results of equipping operational and clinical managers to study problems in their own work settings. These efforts have required no additional internal resources, while the payoffs have been considerable. PMID- 11496477 TI - Queensland public sector nurse executives: job satisfaction and career opportunities. AB - During the past decade, economic and political forces have caused radical transformations in health care systems resulting in changed circumstances within which nursing executives must function. This paper provides an understanding of nursing executives' roles and responsibilities and the impact changes in the health industry have had on their careers. One hundred and forty-seven (52%) of the 281 nursing executives employed in the Queensland Public Health Sector completed a postal self-administered survey. The findings of this study demonstrate their role has expanded to include not only nursing administration, but also responsibility for financial, human resources, strategic and resource management, staff development and quality improvement. The impact of these role changes has affected the health and well-being of nursing executives, with nearly half reporting increased stress, frustration and irritation. Their workload has increased and some reported deterioration in their health, specifically, exhaustion, fatigue and insomnia. Respondents reported they now have less time to spend with families and friends, which has had a negative impact on family relationships. Overall, nursing executives were satisfied with their current position, the work itself and their relationships with their co-workers, but dissatisfied with organisational aspects, especially the quality of mentorship and opportunities for promotion. PMID- 11496478 TI - Health insurance in the Philippines: bold policies and socio-economic realities. AB - In 1995, the Philippines government legislated to create an income-rated and predominantly employment-based universal health insurance program over a 15-year period. The program was intended to provide more and better health care than was available through a combination of existing insurance schemes that covered less than half of the population, and partially subsidized services provided by government facilities and funded from general taxation. The legislation was well intentioned, and the program has some skillful and imaginative staff. However, there are significant barriers to success including low average and widely dispersed incomes, improving but still unsatisfactory health status, weak government health care services, and the sometimes negative impact of for-profit agencies. We review progress to date and conclude that, although membership numbers and benefit rates have increased, access is still inadequate and copayments are high. We argue that strong and innovative steps are needed if the Program's goals are to be realised. In particular, we suggest that the focus should be on more formal and explicit rationing that takes account of cost per quality-adjusted life-year; and radical adjustment of financial incentives for care providers including capitation and per case payment based on costed clinical pathways for high-volume case types. Finally, we comment briefly on lessons that might be learned by both The Philippines and Australia. PMID- 11496480 TI - [Inflammatory pseudotumor of the abdominal cavity]. PMID- 11496479 TI - [Surgical treatment of patients with complications of peptic ulcer of the stomach and duodenum under conditions of a regional hospital]. AB - The authors made an analysis of the incidence and structure of complications requiring reoperations after operative treatment of 176 patients (with a perforated ulcer of the stomach and duodenum--125, with gastroduodenal bleedings- 51). The operative technique for the treatment of different complications is described in detail. PMID- 11496481 TI - [Morphofunctional state of the gastric transplant after surgery of cancer of the cardia and esophagus]. PMID- 11496482 TI - [Choledocholithiasis in middle-aged and aged patients]. PMID- 11496483 TI - [Raynaud's phenomenon: clinical aspects of the disease]. PMID- 11496484 TI - [Dyslipoproteinemia and prognosis of the course of ischemic heart disease after coronary bypass]. AB - The article deals with the observation of 88 patients with ischemic heart disease, 62 of them had coexistent dyslipoproteidemia. More pronounced atherosclerotic lesions of the coronary artery were found to develop in patients with dyslipoproteidemia influencing the degree of ischemic heart disease before coronary bypass. Within the period of 10 years after operation dyslipoproteidemia patients have 3.5 times higher risk of the development of myocardial infarction, more than 2 times higher risk of reappearance of stenocardia and death from cardiovascular diseases. Patients with dyslipoproteidemia have 2 times higher probability of generalization of the atherosclerotic process in patients with is chemic heart disease at later terms after coronary bypass. PMID- 11496485 TI - [Choice of surgical strategy in gastroduodenal ulcer hemorrhage]. AB - An analysis of 567 patients aged from 15 to 89 years with gastroduodenal bleedings was made. Ulcer disease of the stomach was diagnosed in 173 (30.5%) patients, ulcer disease of the duodenum--in 354 (62.4%) patients, 26 patients (4.6%) had combined gastroduodenal ulcers, 14 patients (2.5%) had ulcers of the gastroenteroanastomosis (after Billroth-II operations). Operations were fulfilled in 250 (44.1%) of all the patients. The operation of choice at the high state of bleeding is considered by the authors to be resection of 1/2 of the stomach by the method modified in the clinic. Postoperative lethality was 4%, complications were noted in 12% of the patients. All the complications were nonspecific, i.e. characteristic of any operations on organs of the abdominal cavity. Incompetence of the anastomosis sutures was not noted which in our opinion can be accounted for by a specific method of forming the anastomosis. The average amount of bed days after the operations was 19.5 +/- 3.3. PMID- 11496486 TI - [Endoscopic methods of prevention and treatment of hemorrhage of portal genesis]. AB - Results of endoscopic interventions on 72 patients with portal hypertension of different etiology are presented. In patients with acute bleedings the frequency of primary hemostasis was 94%. In 2 patients with hemorrhages from the cardial veins of the stomach the endoscopic interventions proved to be ineffective. Prophylactic treatment gave 20% recurrent bleedings during a year. Survival of the patients during 6 months was 88.5% and considerably depended on the functional group according to Child. The endoscopic interventions (sclerotherapy, ligation) should be considered effective methods to arrest bleeding and to prevent bleedings of portal genesis. PMID- 11496487 TI - [Classification of colorectal anastomoses]. AB - The authors propose to classify colorectal anastomoses by anatomical signs: high (supraampular), medial (ampular), low (subampular), utmost low (suprasphincteral) and sphincteral. Forty colorectal one row suture anastomoses were performed in patients with carcinoma of the rectum. Incompetent sutures were registered in 3 (7.5%) patients with subampular anastomoses. There were no lethal outcomes. PMID- 11496488 TI - [Content of total immunoglobulin E in patients with acute appendicitis]. AB - The authors investigated the content of total immunoglobulin E in 34 patients with different forms of acute appendicitis. Higher level of immunoglobulin E was revealed in 19 patients. All the patients had destructive forms of appendicitis, in the third of the patients they were complicated by the development of local peritonitis or by spread of the inflammation outside the appendix. The postoperative period in patients with the elevated level of immunoglobulin E was characterized by a higher incidence of purulent wound complications which can be accounted for by the initially more severe forms of lesions of the appendix. PMID- 11496489 TI - [Autovenous bypass in situ in distal occlusions of the arteries]. AB - In patients with bad distal bed and blood flow along the bypass two functioning venous collaterals were left with the diameter less than 1 mm, with the blood flow volume 100-150 ml/min one collateral was left, if the volume was 150-200 ml/min it was not necessary to make an arterio-venous output. One-two functioning collaterals left did not influence the volume and linear velocity of blood flow along the bypass and the parameters of hemodynamics. 78 patients (91%) were discharged with the functioning bypasses and decreased ischemia. A year later thrombosis of the bypass was noted in 6 (14%) of 43 follow-up patients, stepwise liquidation of one or two collaterals was fulfilled in 5 patients due to a more than 1 mm dilatation. Prognostically the blood flow in the zone of a distal anastomosis less than 100 ml/min is thought to be most critical. PMID- 11496490 TI - [Treatment in diffuse phlegmons of the neck]. AB - The authors made an analysis of results of treatment of 146 patients with diffuse phlegmons of the neck. The use of cryoplasmic-antifermentative complex and plasmapheresis in complex treatment allowed to increase the efficiency of pathogenetic therapy for phlegmons of the neck and contact mediastinitis, to improve the entry of antibiotics and enzyme inhibitors into the inflammation focus and to get better results of treatment. PMID- 11496491 TI - [Local application of low-energy aerial and argon plasma in the treatment of suppurative wounds and trophic ulcers]. AB - The authors describe experimental and clinical (113 patients) data demonstrating the antimicrobial, hemostatic and stimulating the regenerating processes property of the low energy plasma in local treatment of purulent and long-standing wounds and trophic ulcers. PMID- 11496492 TI - [Prognosis of early outcomes of severe craniocerebral injuries depending on duration of neurological syndromes and complications]. AB - A mathematical model is presented of prognosis of early (within the periods of stationary treatment) outcomes of a severe cranio-cerebral injury according to the main indicator--"favorable outcome--discharged from the hospital", "unfavorable outcome--died" depending on 10 indicators characterizing the patient's age, injury, duration of neurological syndromes and intraoperative, intracranial postoperative and somatic complications. The model was developed with the help of Excel and the application software Statistica and SPSS for Windows. For the development of the model of the correlative and discriminant analyses were used. The calculation of the prognosis can be made with the help of a computer or a microcalculator. PMID- 11496493 TI - [Cerebrospinal fluid fat globules in traumatic fat embolism]. AB - The article is devoted to the problem of diagnosis and treatment of 28 patients with cranio-cerebral injury with posttraumatic fat embolism. In 15 of them fat liquoroglobulia was diagnosed which is a reliable symptom of the cerebral form of fat embolism. The method of detection of fat liquoroglobulia is described. A new method--liquorosorption--was used in treatment of patients with the cerebral form of fat embolism. A disturbance of the hematoencephalic barrier is thought by the authors to be possible in such embolism. PMID- 11496494 TI - [Chronic recurrent instability of the shoulder joint]. AB - The article gives an analysis of results of conservative treatment of 503 patients with a primary traumatic dislocation of the shoulder. High frequency of the development of chronic instability of the shoulder joint was shown to be 31.2%. The using of arthroscopy, noninvasive methods of diagnosis (rentgenography, USI) allowed to diagnose the typical complex of morphological injuries following dislocations of the shoulder. General and local factors responsible for the development of chronic instability of the shoulder joint were established. Arthroscopy and early surgical stabilization of the joint are thought to be expedient in cases of a combination of these factors. Conservative treatment is indicated to patients older than 40 without high physical needs. PMID- 11496495 TI - [Socioeconomic aspects of the increased volume of cardiosurgical treatment of children with congenital heart defects]. AB - The authors made an analysis of social-economical conditions limiting the possibilities of rendering cardiosurgical care to children. Possible ways are mapped out allowing to increase the amount of operations on children with congenital heart diseases. PMID- 11496496 TI - [Concerning the treatment of patients with inguinal hernia]. AB - The author considers the current state of inguinal herniology. Under analysis are the author's personal materials of many years experiences with surgical treatment of inguinal hernias. An original method of hernioplasty is described which was used in 169 patients. For the results of treatment of hernias to be improved the author proposes and substantiates the necessity of organize herniological Centers with the departments for in- and out-patients. PMID- 11496497 TI - [Immediate results of using the radial artery in direct revascularization of the myocardium]. AB - The authors discuss results of more than 300 operations of the aorto-coronary bypass using the radial artery as a shunt. An analysis of the initial state of the patients, the intraoperative, early postoperative and intrahospital periods was made. Special attention was given to specific features of the isolating and using the radial artery grafts. Specific complications resulting from the application of them were analyzed and recommendations are given for prophylactics of such complications. The experiences with using the radial arteries allow to recommend it in revascularization of the myocardium. It is one of the ways of improving immediate and long-term results of operations on patients with ischemic heart disease. PMID- 11496498 TI - [Surgical treatment of patients with acute abscessed and phlegmonous lactation mastitis]. AB - Under analysis were results of surgical treatment of 256 patients with abscessed and phlegmonous acute lactation mastitis. Radical primary surgical treatment of the purulent cavities allowed prevention of further progression of the inflammation in the mammary gland. The use of the low frequency ultrasound for treatment of the wound allowed to quicker arrest the acute inflammatory process in the tissues of the mammary gland surrounding the purulent cavity. In cases with diffuse purulent acute lactation mastitis it is necessary to take into attention the tension of the suture threads when fulfilling the secondary surgical treatment of the wound and putting in the running secondary early suture. PMID- 11496499 TI - [Use of fibrin-collagen plates in abdominal surgery]. AB - The article presents results of experimental substantiation and clinical use of fibrin-collagen plates Tachocomb. They were used in clinical practice in 28 patients with different bleedings under conditions of open and little invasive surgery. The preparation not only increases the mechanical strength of the surgical suture, but also has a stimulating effect on reparative processes. The author proposes to use Tachocomb for strengthening the sutures and anastomoses in peritonitis, intestinal obstruction and interventions in the area of the tissues altered due to infiltration. PMID- 11496500 TI - [Can perforated ulcer be not sutured?]. AB - Experiments in 5 pigs and 18 mongrel dogs have shown the possibility to close perforating ulcers with plates of biopolymer "Tachocomb". The method is proposed as an alternative to suturing perforations in the zone of pylorus in order to prevent stenosis of the pyloric part of the stomach. The experimental data obtained have confirmed good adhesive and stimulating properties of the fibrin collagen substance. Successful results of using the method were demonstrated in 2 patients with perforating ulcers. PMID- 11496501 TI - [Use of the glue composition TachoComb in forming the portosystemic anastomosis]. AB - The Tachocomb plates were used by the authors in 15 patients for forming the proximal splenorenal anastomosis and selective decompression of the portal system. The data of sonography have shown the valuable functioning of the shunt in all the patients within the period from 6 months to 3 years. The proposed method provides reliable hemostasis during operation and prevents thrombosis of portosystemic anastomoses. PMID- 11496502 TI - [Laparoscopic surgery in tumor obstruction of the biliary tract]. AB - The article gives a detailed description of the technique of laparoscopic biliodigestive anastomoses in patients with advanced tumors of hepatopancreatoduodenal zone. The authors have an experience with 18 operations. They performed 14 cholecystoenteroanastomoses including 9 total laparoscopic operations and 5 operations through a combined approach--laparoscopy in combination with minilaparotomy. The authors believe that laparoscopic surgery is feasible and safe for treatment of these patients. The combined approach has financial advantages giving the possibility to save the disposable stapling devices. PMID- 11496503 TI - [Nikolai Ivanovich Bereznegovskii (1875-1926)]. PMID- 11496504 TI - [Endoscopic removal of the gallbladder mucosa after cholecystostomy]. AB - The authors have developed a new method of radical treatment of patients with acute obstructive cholecystitis after cholecystostomy. The endoscopic removal of the mucosa was proposed using hysteroresectoscope GYI-525-12 (Circon-ACM1). The method was used in 5 elderly patients with high anesthesia risk in whom surgical cholecystectomy was not possible. The removal of the mucosa resulted in the replacement of the gallbladder with the connective tissue giving the effect of radical operation. PMID- 11496505 TI - [Surgical strategy in liver hemangiomas]. AB - The authors made an analysis of results of surgical treatment of 129 patients with hepatic hemangiomas and of the 9-year follow-up of these patients. The dynamic care of these patients is recommended in cases of 5 cm diameter hemangiomas and the absence of clinical symptoms. The surgical treatment was necessary but in 10% of such patients. In the majority of such patients embolisation of the hepatic artery is expedient if surgical treatment is necessary. Indications for resection of the liver are restricted. PMID- 11496506 TI - The new state mental hospitals in the community. AB - Serious problems result when the community alternatives to state hospitalization, often driven by lower costs and an ideology that highly structured care is seldom needed, are not adequate to meet the needs of those who are severely mentally ill. PMID- 11496507 TI - Persons with severe mental illness in jails and prisons: a review. AB - One of the greatest problems of deinstitutionalization has been the very large number of persons with severe mental illness who have entered the criminal justice system instead of the mental health system. PMID- 11496508 TI - Deinstitutionalization at the beginning of the new millennium. AB - This overview of deinstitutionalization explores its accomplishments and disappointments. PMID- 11496509 TI - Community treatment of severely mentally ill offenders under the jurisdiction of the criminal justice system: a review. AB - Community treatment of severely mentally ill offenders who fall under the jurisdiction of the criminal justice system has important differences from treatment of non-offenders. It is critical to identify a treatment philosophy that strikes a balance between individual rights and public safety and includes clear treatment goals. PMID- 11496510 TI - Outcome for psychiatric emergency patients seen by an outreach police--mental health team. AB - Outreach emergency teams comprising police officers and mental health professionals can help to avoid criminalization of the mentally ill. PMID- 11496511 TI - Court intervention to address the mental health needs of mentally ill offenders. AB - Mental health consultation is provided to a municipal court that recommends court mandated interventions for mentally ill persons who have committed minor crimes. This study demonstrates that a significantly better outcome results when the judge not only mandates but monitors mental health treatment. PMID- 11496512 TI - Therapeutic use of conservatorship in the treatment of gravely disabled psychiatric patients. AB - Mental health conservatorship can play an important role in the clinical management and treatment of persons with severe mental illness. PMID- 11496513 TI - A century and a half of psychiatric rehabilitation in the United States. AB - After a brief history, this chapter describes important modalities and models of psychiatric rehabilitation. PMID- 11496515 TI - Disaster planning. PMID- 11496514 TI - Accurate heights and weights. PMID- 11496516 TI - Management of the environment of care (EC). PMID- 11496517 TI - [The state of cancer surgery at the turn of the millennium]. AB - Surgery remains the method of choice for the treatment of stage I and II cancer disease, it has proved to be curative in 40% of these cases. The role of surgery in the complex multimodality treatment of tumorous diseases is changing, depending on the type and stage of the disease and on the effectivity of the adjuvant therapeutical modalities. In general, surgery has not lost its significance in the treatment of cancer during recent decades. The author presents precise definitions and a homogeneous terminology for the basic concepts of oncological surgery. Changes in the surgical strategy and other factors influencing the surgical radicality are analysed. New methods contributing to the development of cancer surgery and to the improvement of the survival rate and the postoperative quality of life are surveyed. The roles of regional lymphadenectomies, metastasectomies, extended resection and organ-saving methods in the up-to-date treatment of tumorous diseases are discussed. PMID- 11496518 TI - [Poliomyelitis and the post-polio syndrome]. AB - In developed countries as well as in Hungary polio virus related disease disappeared completely due to the extensive administering of vaccine. As a result, young and middle-aged doctors have no experience of encountering acute polio virus infection but instead they meet its resultant impairments and disabilities. Persons who had suffered the onset of poliomyelitis 3 or 4 decades earlier, may to develop a new set of symptoms and functional declines. The criteria for post-polio syndrome were identified and its impairments and disabilities were described. It is only within the last decade that the adaptive changes in muscular tissue, insufficiently compensed denervation, together with the effects of physiotherapy, have become better known. The aim of this paper is to review the present situation concerning the struggle for keeping the polio virus in check, to give a short summary of the post-polio syndrome and to draw attention to the importance of rehabilitation. PMID- 11496519 TI - [Significance of anamnestic factors in the background of pathologic airway alterations]. AB - The results of 867 diagnostic bronchoscopies, between August 1992 and August 1993, were studied by the retrospective analysis of the patients' files. The average age of the children was 46 months (6 days-25 years). The proportion of girls and boys were 38.6% and 61.4%. Data of the history were: therapy resistant recurrent wheezy bronchitis in 31.9%, recurrent pneumonia in 31.1%, stridor in 22.1%, recurrent croup in 17.4%, bronchial asthma in 10.1%, monosymptomatic cough in 9.5% and recurrent bronchitis in 8.0%. The 768 (89%) pathologic findings were: 1. by bronchoscopy (n = 867): acute inflammation of the mucous membrane 7.3%, chronic bronchitis 75.1%, mucous secretion 44.5%, purulent secretion 20.8%, cricoid stenosis 4.6%, tracheal stenosis 11.2%, left main stem bronchus stenosis 27.8%; 2. by bronchography (n = 202): bronchial deformation 43.1%, bronchiectasis 5.4%, gracile bronchi 2.0%. The following significant relations were proved between the anamnesis and the results of the bronchoscopy and/or bronchography by statistical analysis: recurrent wheezy bronchitis/chronic bronchitis (p < 0.001; chi 2 = 16.35), approximately/purulent secretion (p = 0.039; chi 2 = 4.26), approximately/left main stem bronchus stenosis (p < 0.001; chi 2 = 19.27); stridor/tracheal stenosis (p < 0.001; chi 2 = 58.67), approximately/left main stem bronchus stenosis (p < 0.001; chi 2 = 63.45), approximately/cricoid stenosis (p = 0.0015; chi 2 = 10.8), approximately/other stenosis (p = 0.002; chi 2 = 9.69); recurrent croup/cricoid stenosis (p < 0.001; chi 2 = 18.34); bronchial asthma/bronchial deformation (p = 0.026; chi 2 = 4.95). The results show the importance of the bronchological examination in the diagnosis of chronic chest symptoms in children. PMID- 11496520 TI - [Percutaneous sclerotherapy of thyroid cysts]. AB - The authors have performed percutaneous ethanol sclerotherapy in 9 patients suffering from thyroid cysts causing physical and cosmetic complaints during the last 4 years. All cysts proved to be benign by US and aspiration cytology. The sclerotherapy resulted in definitive recovery in 8 cases. No complications of the procedure were noticed. PMID- 11496521 TI - [75 years of Hungarian psychiatric care]. PMID- 11496522 TI - [Andras Fay, academician, life insurance expert]. PMID- 11496523 TI - [Dr. Jozsef Fodor about longevity--death centenary of the professor-academician]. PMID- 11496525 TI - Testing your diagnostic skills (#52). Case 1. Metal tongue stud. PMID- 11496526 TI - Testing your diagnostic skills (#52). Case 2. Odontogenic keratocyst. PMID- 11496527 TI - Americans with Disabilities Act affects dental profession. PMID- 11496528 TI - Class-action racketeering lawsuit filed against medical HMOs. PMID- 11496529 TI - New law could change scope of practice, standards of patient care. PMID- 11496530 TI - Alternative and complementary health care treatments. PMID- 11496531 TI - Of mice and mongooses ... a history of leptospirosis research in Hawaii. AB - A history of leptospirosis research in Hawaii is presented, beginning with the first published work in 1937. This account traces the leading researchers who described the organism and the disease, the diagnostic tests developed and used, the reservoir animals identified, methods of disease transmission discovered, prevention programs developed in the state, and research into more effective disease detection and prevention. PMID- 11496532 TI - Teaching cultural competency. PMID- 11496533 TI - Quality of life research in Hawaii's cancer survivors. PMID- 11496534 TI - [Diabetic nephropathy]. PMID- 11496535 TI - [Pre-occupying increase in diabetes as cause for terminal kidney failure. Evaluation of treatment strategies]. AB - OBJECTIVE: To study the incidence, prevalence, treatment modalities and prognosis of diabetic patients on renal replacement therapy. MATERIAL: We studied all end stage renal failure (ESRF) diabetic patients on renal replacement therapy in "Area Sanitaria 1" in Madrid from 1978 to 1998. RESULTS: Diabetes mellitus has become the leading cause of ESRF in our health area of Madrid, 30% of all causes of ESRF. Incidence of diabetics beginning RRT was 33.3 per million population (pmp) in 1998, while in 1993-94 diabetes was 15% and 21 pmp, respectively, table 1. The proportion of diabetics on RRT has increased from 7.4% 1986 to 12.7% 1998. The prevalence of diabetics on RRT in 1998 was 135 pmp, with an overall prevalence of 1054 pmp. At the mean time, the proportion between incident diabetics type 2/diabetics on RRT has increased from 15% in 1987-88, to 54% in 1993-94 and to 81% in 1997-98, consequently, mean age of diabetic patients at beginning RRT has increased from 47 years before 1988, 58 in 1989-90, 61 in 1993 94, 62 in 1995-96 and 63 in 1997-98 (Table I). We studied 182 diabetics admitted for renal replacement therapy, 106 males and 76 females, 105 were diabetics type 1 and 77 type 2. Their mean age at RRT beginning was 57.12 years (SD). Hemodialysis (HD) was the first modality of treatment for 128 (70%) diabetics and CAPD for 54. Seventy out of 128 patients on HD were dialyzed in the Hospital Unit, 40% on AFB (acetate free biofiltration) and 58 in two Satellite Units, that means a higher proportion of diabetics on CAPD and on HD in Hospital Unit. Diabetics allocated to CAPD were 5 years, as mean, younger than patients allocated to HD (p < 0.01) and the proportion of diabetes type 1 was higher in CAPD (72%) than in HD (51%), p 0.05. During the mean follow-up period (51.45 months) 79 patients changed their treatment modality and 45 of them received a kidney allogral. Relative risk of drop-out was higher in CAPD technique when compared to HD. Clinical complications were frequently observed: ocular (77%), cardiovascular (Myocardial infarction 1.7%), acute cerebrovascular disease (21%) and distal angiopathy (35%), 10% amputee. At December of 1998, 89 patients were dead. Cardiovascular and cerebrovascular diseases (29%) and Infections (27%) were the two most frequent causes of death. Unknown-origin deaths represent 19% of all deaths. The overall survival (Kaplan Meier) was 92%, 80%, 72%, 61% and 54% at 1, 2, 3, 4 and 5 years, respectively (57 patients completed last period). Survival was better on HD than on CAPD, but without statistical significance, although the significant difference in age and in proportion of diabetes type 2 between both groups. Data analysis estimated by Cox proportional hazards regression shows that younger age and ki,r transplantation had a positive independent effect on survival, whik clinical distal angiopathy had important negative effect on survival. CONCLUSIONS: Diabetes mellitus has become the leading case of ESRF in our health area of Madrid. Young age and kidney transplantation had a positive independent effect on survival, while clinical distal angiopathy had important negative effect. PMID- 11496536 TI - [Fever of unknown origin in HIV positive patients]. AB - OBJECTIVES: To describe the clinical presentation of FUO in patients infected with HIV and to asses the diagnostic usefulness in this population of several test usually recommended in the study of FUO. PATIENTS AND METHODS: We studied the clinical charts of all patients with HIV that required admission to our Hospital during a 23-month period. FUO was defined according to Petersdorf's modified criteria. Fifty-four patients fulfilled study criteria during the entry period. RESULTS: A cause of fever was identified for 48 patients (89%). Tuberculosis, disseminated atypical mycobacteriosis and Leishmaniasis can explain 68% of them. Examination of bone marrow aspirates, lymph node aspirates and biopsy, and culture of clinical specimens for mycobacteria were the procedures with the highest diagnostic yield. CONCLUSIONS: Mycobacterial infection should be considered as a first-line diagnosis in HIV-patients with FUO. It is possible to predict the diagnosis of tuberculosis infection with a high level of confidence (90.5%) through a logistic regression model based on easily obtainable parameters. PMID- 11496537 TI - [Factors that influence the non prescription of beta-blockers in patients with acute myocardial infarction]. AB - OBJECTIVE: To study the factors that influence the non prescription of beta blockers in patients discharged with a diagnosis of acute myocardial infarction (AMI). METHOD: A retrospective study was done of all patients discharged from our Service in the year 1998, with a diagnosis of AMI. The variables considered were age, sex, diabetes mellitus, peripheral vascular disease, left ventricle dysfunction and COPD. RESULTS: 60 patients with AMI were included in the study, 18 of whom (30%) were discharged without beta-blockers. The average age of these patients was 77 years, while the average age of those discharged with these pharmaceutical agents was 60 years (p < 0.0001). Likewise, left ventricle dysfunction (p < 0.031) and female gender (p < 0.016), also negatively influenced the use of these drugs. It was observed with multivariable regression analysis that age was the main predictor for the use of beta-blockers (p < 0.0001). CONCLUSION: Age is the main factor that influences the non prescription of beta blockers in patients with AMI in our Service. In spite of the potential adverse effects of the drugs in the advanced age population, the data so far obtained demonstrates a clear benefit in the subgroups at risk (advanced age, heart failure,...). Therefore it's use should be extended to this group of population as long as there are no absolute contraindications. PMID- 11496538 TI - [Giant cell arteritis and polymyalgia rheumatica: diagnosis and evolution of 90 cases]. AB - OBJECTIVE: To compare the clinical features of giant cell arteritis (GCA) and polymyalgia rheumatic (PMR), and to evaluate the criteria proposed by the American College of Rheumatology in our results. PATIENTS AND METHODS: A retrospective analysis of 90 patients with GCA (n = 57.55 of whom were diagnosed by biops) and PMR (n = 33) diagnosed over the last 10 years. RESULTS: Headache was present in 45 patients (78.94%) with GCA and in 7 (21.21%) with PMR (p < 0.001); polymyalgic syndrome was observed in 15 patients (26.31%) with GCA and in 33 (100%) with PMR (p < 0.001); jaw or tongue claudication was observed in 14 patients (24.57%) with GCA and in 2 (6.06%) with PMR (p < 0.05), and visual disturbances were only present in 9 patients (15.79%) with GCA. The erythrocyte sedimentation rate (ESR) was > or = 50 mm/h in 84 patients (93.33%), and > or = 100 mm/h in 43 of them (51.19%). The ESR became normal (20 Pounds mm/h) in less of 8 weeks after the treatment was started in 64 patients (76.19%). At the time of diagnosis, 61 patients (61.67%) had anemia, which was severe (Hb < 10 g/dl) in 17 cases (27.86%). After steroid treatment 43 patients (70.49%) improved their anemia in less of 12 weeks, and 25 of them (58.13%) in less of 8 weeks. CONCLUSIONS: The cranial symptoms were predictive for a positive temporal artery biopsy. The anemia and its quick normalization after steroid treatment can help to the diagnostic. PMID- 11496539 TI - [The retinoic acid syndrome, a complication of acute promyelocytic leukemia therapy]. AB - OBJECTIVE: To describe the retinoic acid syndrome, a complication of therapy with all-trans retinoic acid (ATRA) in acute promyelocitic leukemia (APL). PATIENTS AND METHODS: Retrospective study of five patients with a morphologic diagnosis of APL by the French-American-British (FAB) classification that were treated for remission induction with ATRA and developed the ATRA syndrome. RESULTS: Three patients in newly diagnosed APL and two in leukemia relapse were analyzed. All patients received with ATRA 45 mgrs/m2/day, and three of them also received chemotherapy. Patients developed fever, respiratory distress, pulmonary infiltrates, weight gain and edemas. The onset of this symptom complex occurred from 1 to 11 days after starting treatment and was preceded by an increased in peripheral blood leukocytes. Infections or congestive heart failure were ruled out. The clinical course progressed while patients being treated with wide spectrum antibiotics. Four patients were treated with high doses corticosteroid therapy (dexametasone 10 mgrs intravenously every 12 hours), in three of them full recovery was attained and one died. One patient that did not received steroids died. CONCLUSIONS: The use of all-trans retinoic acid to induce hematologic remission in APL patients is associated in same patients with the development of ATRA syndrome, a life threatening complication. Symptoms begin in the first days of treatment. If this syndrome is suspected, early treatment with high dose steroids should be initiated. PMID- 11496540 TI - [Cholesterol crystals embolization. A case report]. AB - Cholesterol Crystals Embolization is not a very frequent entity that happens in people with atheromatous disease of spontaneous for or after the realization of vascular manipulation (angiographic, angioplasty procedures, cardiovascular surgery) or more rarely with the treatment with oral anticoagulants and after thrombolytic therapy. The clinical presentation is according to the arterial affected territory and so from diagnosed cases in autopsy reports to florid clinic forms exist. The more frequent are the cutaneous manifestations with apparition of livedo reticularis and purple toes that could go accompanied of necrosis of lower limbs, consecutive from the renal manifestations and the gastrointestinal tract one. The diagnosis is based on the biopsy of organ affected lesions to level of arterioles, that revealed characteristic cholesterol clefts, above all the cutaneous biopsy, muscular and in any case renal. As for the treatment is symptomatic with analgesics, antiplatelet agents, anti inflammatory, etc ... but it doesn't after the presage and the evolution of the illness. A clear contraindication exists to the use of oral anticoagulants and heparin. PMID- 11496541 TI - [Primary systemic amyloidosis presenting as polymyalgia rheumatica and giant cell arteritis]. AB - Primary systemic amyloidosis or AL-amyloidosis is an uncommon disease characterized by the accumulation in vital organs of a fibrillar protein consisting of monoclonal light chains. It is a plasma-cell dyscrasia related to multiple myeloma where clonal plasma cells in the bone marrow produce immunoglobulins that are amyloidogenic. A monoclonal component is present in the serum or urine of 90% of patients. The presentation of most patients with AL amyloidosis is usually related to congestive heart failure, nephrotic syndrome o peripheral neuropathy, but there are unusual features suggesting giant cell arteritis (GCA) and polymyalgia rheumatic (PMR). Although in the majority of AL cases the plasma cells clone is small, the assumption is that the outcome of the disease is uniformly fatal (median survival 12-15 months) and treatment is analogous to those used in malignant proliferative disease. We describe a patient with AL amyloidosis who presented with manifestations of GCA and PMR, and we review the main characteristics of primary amyloidosis. PMID- 11496542 TI - [Paraquat poisoning: report of two cases and literature review]. AB - Cases of poisoning with pesticides, especially suicidal ones, continue to be an important therapeutic problem. The heribicide paraquat (1.1' dimethyl-4.4' bipyridylium dichloride) is the second cause of pesticide poisoning in our country, which is associated with a high mortality rate. We report two cases of suicidal ingestion of paraquat who developed multiorgan failure with a lethal outcome. We also present a brief review of the literature, mainly focused on the different therapeutic options. PMID- 11496543 TI - [Homocysteine. The cardiovascular risk factor of the next millennium?]. AB - Several epidemiologic studies have demonstrated that hyperhomocysteinemia is a risk factor for arteriosclerosis in coronary, cerebral, peripheral and aortic arteries. This risk is independent of other cardiovascular risk factors, and it is dose related. However, prospective studies show contradictory findings. Hyperhomocysteinemia is also associated with a higher risk of venous thrombosis to which other coagulation disorders, such as factor V Leiden, could contribute. Hyperhomocysteinemia can be due to genetic defects in the enzymes that control homocysteine metabolism, and also to other factors, mainly nutritional (deficiencies in vitamin B6, vitamin B12, or folic acid). Dietary supplements of these vitamins reduce plasma homocysteine levels. Randomized clinical trials are still needed to demonstrate that reducing plasma homocysteine levels will reduce the risk for cardiovascular disease. PMID- 11496544 TI - [Neurological manifestations of systemic lupus erythematosus. A challenge for the diagnosis]. PMID- 11496545 TI - [Myocardial ischemia after a wasp sting]. PMID- 11496546 TI - [Primary pericardial lymphoma in HIV infection]. PMID- 11496547 TI - [Leptospirosis in immunocompetent patients]. PMID- 11496548 TI - [Bronchogenic cyst extended to the esophagus]. PMID- 11496549 TI - [Gouty arthritis in young patient]. PMID- 11496550 TI - [Antral vascular ectasia as a cause of ferropenic anemia]. PMID- 11496551 TI - [Peritoneal mesothelioma in a man who transported asbestos]. PMID- 11496552 TI - [Acute hypoglycemia as first manifestation of a Burkitt-like lymphoblastic leukemia]. PMID- 11496553 TI - [Autoimmune hemolytic anemia and hypernephroma]. PMID- 11496554 TI - [Association of giant cell arteritis and hearing loss]. PMID- 11496555 TI - [Acarbose and diabetes mellitus. Practical implications]. PMID- 11496556 TI - [Changes in metabolic parameters and microalbuminuria in patients with type 2 diabetes treated with acarbose]. AB - OBJECTIVE: To determinate the influence of the acarbosa and microalbuminuria on metabolic parameters in patients diabetics type 2. RESEARCH DESIGN AND METHODS: We are studied 92 patients with diabetes type 2, treated with Acarbose alone or with insuline or oral anti-diabetics We are determined the values of HbA1c, total cholesterol, HDL, LDL, triglycerides and microalbuminuria, before and after or the treatment with acarbose. RESULTS: The patients presented globally a statistically significant improvement in the levels of Hb A1 c, triglycerides and microalbuminuria. In the group treatise with 300 mg/day of acarbose it was observed a significant decrease of the values of Hb A1c and triglycerides independently of the associated treatment. In the group treated with 150 mg/day the improvement only it was statistically significant for the levels of Hb A1c. CONCLUSIONS: The treatment with acarbose produces an improvement in the levels of Hb A1c independently of the administered dose and of the triglycerides in patients treated with 300 mg/day without relation to associated treatments. Therefore, he acarbose win be considered an effective medication to improve the metabolic control to prevent the cardiovascular risk in those patients. PMID- 11496557 TI - [Pulmonary diffusion of carbon monoxide in 2 clinical situations: bronchial asthma and diabetes mellitus]. AB - OBJECTIVE: The aim of our work has been the study and comparison of diffusion capacity of CO (DLCO) in two different clinical situations: bronchial asthma and diabetes mellitus. METHOD: We have studied 16 control subjects, 38 patients with bronchial asthma and 65 patients with diabetes mellitus. We performed CO pulmonary diffusion tests by single breath method to determine two components: membrane diffusion factor (Dm) and pulmonary capillary blood volume (Vc). RESULTS: We have found a positive correlation of FEV1 with Dm. The bronchial asthma group had a lower FEV1 and FEF25-75% and an increase in DLCO, Dm and Vc, with respect to the control group. The diabetes mellitus group presented a decrease in CVF, FEV1, DLCO and Vc, with respect to the control group. The bronchial asthma group showed a lower ratio of Dm/Vc than the control and diabetes groups. CONCLUSIONS: The bronchial asthma patients have an increase in CO pulmonary diffusion, membrane diffusion factor and pulmonary capillary volume. However, the diabetes mellitus patients present a decrease in CO pulmonary diffusion mainly due to pulmonary capillary volume. PMID- 11496558 TI - [Recurrence of tuberculosis in patients infected with the human immunodeficiency virus]. AB - OBJECTIVE: To determine the clinical manifestations of the relapse of the tuberculosis (TB) in the patients infected by the human immunodeficiency virus (HIV) and the degree of immunodeficiency shown. MATERIAL AND METHOD: They are analyzed retrospectively 121 cases, all they HIV+, diagnosed of TB from November of 1985 until December of 1999. All they fulfilled correctly the specific treatment of the tuberculosis. RESULTS: Of 121 studied cases, a total of 13 (10.7%) presented relapse of tuberculosis finally of a mean of 31.8 months (range 1-95). In 7 cases the affectation was pulmonary (2 typical, 5 non typical), in 3 mixed (pulmonary and extrapulmonary) and in 3 extrapulmonary. The clinical sintomatology were present in 92% of the cases and, predominantly, was: fever (92%), cough (69%), sweating, weight loss (61%). Hepatomegaly results be the exploratory finding quite frequent (54%). The mean of CD4+ lymphocytes was 141.7 (range 3-406), in 8 (61%) the lymphocyte counts was inferior to 100 cells/mm3. PPD was positive in 1 case (14%) and with respect to delayed-type hypersensitivity reaction test, it was observed not normoergia in all the cases (100%). CONCLUSIONS: Relapse of the tuberculosis in patient infected by the VIH is presented with a manifested clinical expresivity in the course of a severe degree of immunodeficiency. In certain groups of risk as the intravenous drugs users, it must of be had a high level of suspicion of the relapse of the disease, above all in those patients that show low CD4+ lymphocyte counts. PMID- 11496559 TI - [Hospital readmission in internal medicine]. AB - BACKGROUND: The readmission rate could be a valuable tool as measurement of hospital quality. Readmissions are due to several factors: clinical, hospital related and patient related. We analyze readmission to internal medicine in a hospital of third level. MATERIAL AND METHODS: During 11 months in 1988 we counted all readmissions (R) defined as every previous admission occurred in a span of five years into an area of internal medicine composed by 8 beds. We counted number of readmssions, time from the last readmission, living area (city vs country), sort of primary care physician (GP vs family care specialist), living way (single, with family, institution, homeless). Precipitating factors were observed as well as diseases causing it. R were classified as R related (RR) when readmission was provoked by the same pathological condition or a complication. Multi-readmission (MR), those R caused by the same disease process and treated in different areas and services of the hospital. Avoidable R (AR), those R which did not fullfil AEP criteria. Early readmission (ER) those R occurring before 30 days after last discharge. RESULTS: Three hundred and eleven patients (mean age 67.93 (SD 15.51), males 64%, mean length of stay 7.75 (SD 4.35), 93% admitted from emergency yard, mortality rate 3.5%) were included. R were 111 (35.5%), RR 83 (26 and 75% of RR), MR 68 (82% of RR), ER 33 (39.7% of RR) and AR 16 (19.2% of RR) patients. The most frequent diseases were heart failure and chronic respiratory diseases. Main causes of R were worsening of chronic disease 41 (37%), non-appropriale ambulatory management 24 (22%) erroneous diagnosis 8 (7%), iatrogenic effect 7 (6%), new disease 29 (26%) and others 2 (2%). Mortality rate in R patients was 7.2% (confidence interval 95% 2 to 9%). Number of readmissions were 3.22 (SD 2.25) and time to readmission 8.99 (SD 11.96) months. Living in city (p < 0.05) and to be cared by family physician (p < 0.01) both were factors accelerating readmission. Patients with RR had a higher number of readmissions (3.55 SD 2.23 p < 0.001) and they occurred sooner (8.03 SD 11.85) (p < 0.01). There was a trend to higher readmission rate in female (p 0.052). Fifty-seven percent of RR patients did not have consultation with primary care physician (p < 0.05) (confidence interval 95% 3 to 39%). Consultation with primary care yielded a delay in readmission of 5 months (p < 0.01). Patients with MR had an increased number of readmissions (p < 0.01). Associated factors were iatrogenic effect (p < 0.05), non-appropriate ambulatory management (p < 0.001) and worsening chronic disease (p < 0.001). Patients with ER were readmitted 0.45 (SD 0.30) months after the last discharge and they had a higher mortality rate (p < 0.05). Patients with AR had a mean length of stay shorter (p < 0.05), a trend to higher readmission rate (p = 0.06) and sooner (p = 0.08) with a null mortality rate (p < 0.01). As risk factors for RR in logistic regression were identified MR, AR, ER and causes of readmission consisting in worsening of chronic disease, non-appropriate ambulatory management, erroneous diagnosis and iatrogenic effect. CONCLUSIONS: Our readmission rate is 26%, chronic respiratory diseases and heart failure being the main diseases. Over 39% of causes of readmission could be preventable and there is a facilitation phenomenon in number and time to readmission caused by previous readmissions. Risk factors for readmission in internal medicine are multi-readmission, early and avoidable readmission and as specific causes worsening of chronic disease, non-appropriate ambulatory management, erroneous diagnosis and iatrogenic effect. PMID- 11496560 TI - [Bartonella henselae endocarditis. Report of a case and review of the literature]. AB - Bartonella spp are small Gram-negative rods, aerobic and highly fastidious. They are difficult to culture, in the routine bacterial cultures. They are considered as emergent human pathogens. Since 1993, three species of Bartonella (B. quintana, B. henselae, and B. elizabethae) have been described as causative agents of infectious endocarditis. In this paper we describe the case of a 43 year-old woman with a previous valvular heart disease, probably of rheumatic origin, owner of cats, that suffered an infectious endocarditis by Bartonella henselae in the aortic valve. This patient presented IgG titers against B. henselae of 1/4096 and against B. quintana of 1/256. She also had low IgM titers against B. henselae and B. quintana: 1/64 and 1/32, respectively. The patient received antibiotics for 12 weeks and suffered a valvular replacement due to the severe lesion on the aortic valve. On the endocardiac tissue of the removed valve DNA of B. henselae was detected (polymerase chain reaction-based assay). Clinical evolution of the patient was good. Diagnosis of Bartonella spp infection must be considered in every patient with infectious endocarditis and negative blood cultures, and particularly B. henselae in patients with previous valvular heart disease and regular contact with cats. PMID- 11496561 TI - [Efficacy of quinagolide in the treatment of a patient with hypophyseal resistance to thyroid hormones]. AB - The pituitary resistance to thyroid hormones (PRTH) is not very frequent and well known entity, their treatment it continues being topic of controversy. In this work we have evaluated the quinagolida effectiveness in the treatment of it unites patient with (PRTH). The relationship among thyroid stimulating hormone (TSH) and free triiodothyronine (FT3) it was used as marker of the thyroid resistance and of the response to the treatment. The concentrations of TSH and FT3 were normalized after adding quinagolida to methimazole. These results suggest that the quinagolida could be an useful drug in the treatment of this pathology, next to the classic treatments. PMID- 11496562 TI - [Hyperthyroidism, jaundice, and pulmonary hypertension]. AB - Although the most frequent cause associated to the alteration of hepatic function tests that some hyperthyroid patients show is concomitant heart insufficiency, some cases without heart insufficiency and normalisation of hepatic tests after normalising the thyroid function have been described. There is also some evidence of hyperthyroid patients with pulmonary arterial hypertension, with no clear cause, coming back later to normal figures or substantially descending, after the treatment for hyperthyroidism. We show the concrete case of a 66 years woman, who has a due to toxic multinodular goiter hyperthyroidism with jaundice and serious pulmonary hypertension associated, unexplainable by further causes, both regressing with normalisation of thyroid function with no evidence of associated autoimmune pathology. Following, the physiopathology explanations about this unusual association found in literature, are commented. PMID- 11496564 TI - [Exercise-induced anaphylaxis syndrome]. AB - In the last two decades a growing incidence of a peculiar form of anaphylaxis that only occurs while carrying out physical exercise has been observed. Within the exercise-induced anaphylaxis syndrome two well differentiated clinical forms are included: systemic cholinergic urticaria and exercise-induced anaphylaxis in the strict sense which can be shown by a classic form or a variant form, more uncommon and with manifestations similar to cholinergic urticaria. Postprandial or food-dependent exercise-induced anaphylaxis is a frequently identified subtype of these last cases. It can be due to an asymptomatic food allergy manifested through physical effort, although in many occasions it is not possible to find a responsible allergen. The diagnosis is settled on the clinical history and specific tests with food allergens. It can be necessary to perform an exercise challenge test with and without previous ingestion. The treatment is preventive and it is based on avoiding the food or the food allergen some hours before the exercise. When it does not depend on foods it is used a prophylactic pharmacotherapy with antihistamines, cromones or sodium bicarbonate. The patient should be well educated on the use of epinephrine in the event of new reactions. PMID- 11496563 TI - [Acute water intoxication as complication of intravenous urography]. AB - Physiologically, two complementary mechanisms regulate plasma osmolality: antduretic hormone ADH) and thirst. ADH release s supressed, thirst s inhbted and renal water loss occurs when plasma osmolality below a threshold level. The rise in plasma osmolalty causes ADH release, stimulation of thirst and water intake. Acute water intoxication is exceptional in patients without a chronic psychiatric disease. Herein, we describe a case of acute water intoxication in a previously healthy patient, after making an intravenous urography. The excessive water intake and the impossibility of renal water loss because of streee-induced ADH release originated t. Only nine cases have been previously described; almost they all were women preparing for diagnostic procedures. PMID- 11496565 TI - [Sleep apnea syndrome and cardiovascular diseases]. AB - The sleep apnea syndrome is a common disease, recognised as a public health problem. Cardiovascular disease is the most frequent cause of morbidity and mortality in these patients, however the underlying mechanisms of this association have not been clearly established. In sleep apnea syndrome different phenomena can be produced which may explain the appearance of cardiovascular problems, such a progressive hypoxia in relationship with the apnea, the increases of intrathoracic pressure cause by the efforts of ventilation system against close upper airway and the modifications of the autonomic nervous system associated with the arousals. In addition, the hypoxia episodes and reoxygenation, which appear in the sleep apnea syndrome, may play a important role in the alteration of the balance between vasoconstriction and vasodilatation substances affecting the vascular homeostasis and conditioning endothelial dysfunction. On the other hand, the increasing of platelets aggregation and the decreased of fibrinolisis in this group of patients may cause vascular diseases. PMID- 11496566 TI - [Fascioliasis in the south of Galicia. Report of 2 cases]. PMID- 11496568 TI - [Sporadic Pontiac fever]. PMID- 11496567 TI - [Agranulocytosis and ticlopidine]. PMID- 11496569 TI - [Diarrhea as initial manifestation of immunoglobulin B producing intestinal immunocytoma]. PMID- 11496570 TI - [Systemic mastocytosis and magnetic resonance]. PMID- 11496571 TI - [Etiopathogenesis of hereditary periodic fever syndromes]. PMID- 11496572 TI - [Hypergammaglobulinemia D syndrome]. PMID- 11496573 TI - [Consent and confidentiality in medical practice in AIDS]. PMID- 11496574 TI - [Pulmonary nodular sarcoidosis]. PMID- 11496576 TI - I'm 60-70% ok, you're 60-70% ok. Clients don't seek perfection in counseling experience. PMID- 11496575 TI - [Tumor markers in lung cancer]. PMID- 11496577 TI - ECT: the debate returns. PMID- 11496578 TI - Cutting edge science and spirituality: can they co-exist? PMID- 11496579 TI - Serving youths by modifying treatment. PMID- 11496580 TI - A new direction for managed behavioral healthcare. PMID- 11496581 TI - From police officer to care partner. PMID- 11496582 TI - Understanding Rx options for the youngest patients. PMID- 11496584 TI - Weighing options in emergency psychiatry. PMID- 11496583 TI - Does the alcoholic's remedy come in a pill? PMID- 11496585 TI - Recovery of Medicare and Medicaid overpayments in bankruptcy. AB - Mr. Roest argues in favor of the Third Circuit's decision in University Medical Center v. Sullivan to advance the proposition that the Doctrine of Recoupment should be applied narrowly in health care bankruptcy cases. The article begins by introducing key provisions of the Medicare Act and Bankruptcy Code, and by distinguishing between recoupment and setoff. The article then focuses on the Third Circuit's decision, giving both a sketch of the court's decision and a commentary on the holding. The article concludes with a discussion of recoupment related issues left open by the University Medical Center decision. PMID- 11496586 TI - Regulation of online pharmacies: a case for cooperative federalism. AB - Ms. Zeman examines the regulatory challenges and responses arising from online pharmacies. In particular, Ms. Zeman discusses the roles taken by the States' attorney general offices, the States' legislatures, and the pharmaceutical industry itself, to ensure protection for those consumers seeking health care via the Internet. PMID- 11496587 TI - Internet prescribing limitations and alternatives. AB - Ms. Friedman discusses the legal limitations placed on the consumer's ability to obtain drug prescriptions via the Internet. The article focuses on current legislative policies and regulations used to restrict access to prescription drugs over the Internet in the absence of a well-founded physician/patient relationship. The article argues that regulators might be able to satisfy the policy objective of ensuring that a learned intermediary is involved in prescription drug therapy by expanding the prescriptive authority of licensed pharmacists. PMID- 11496588 TI - The changing role of pharmacy practice--a clinical perspective. AB - Professor Carmichael and Dr. Cichowlas argue for the reexamination of the pharmacist's role in the current health care system. Reexamination is necessitated due to changing complexities of the health care system as evidenced by increased economic demands and pharmaceutical surpluses. The authors advocate for Collaborative Drug Therapy Management, in which physicians and pharmacists maximize patient care by pooling their areas of expertise together. PMID- 11496589 TI - The government, the legislature and the judiciary--working towards remedying the problems with the civil False Claims Act: where do we go from here? AB - Ms. McClain examines the government's role in developing and applying the False Claims Act. The article explores the use and effectiveness of the False Claims Act to target fraud and other abuses in the health care system. The article then addresses past inequitable misuse of the Act resulting from improper judicial interpretation of the Act and offers suggestions for curbing some of the stricter provisions of the Act. PMID- 11496590 TI - Striving for a secure environment: a closer look at hospital security issues following the infant abduction at Loyola University Medical Center. AB - Ms. Goodwin discusses the need for heightened hospital security in light of the infant abduction at the Loyola University Medical Center in May, 2000. The article first discusses general security issues endemic to the hospital environment; it then explores protective measures enacted by legislative agencies to reduce the likelihood of violence in hospitals. The article then points to recent litigation arising out of a failure to create a safe hospital environment. Finally, Ms. Goodwin considers the consequences that hospitals and society must face when the hospitals fail to comply with safety procedures. PMID- 11496591 TI - Responsive regulation of Internet pharmacy practice. AB - Professor Brushwood discusses the effectiveness of the Internet as a medium for carrying out pharmaceutical care. A proponent of Internet pharmacy, Professor Brushwood argues that pharmacy regulators could best protect and promote public health through responsive Internet regulation. Wary of state paternalism, the article advocates the Verified Internet Pharmacy Practice Site program of the National Association of Boards of Pharmacy as a model method for regulating pharmacy practices over the Internet. PMID- 11496592 TI - [Renal transplantation and pregnancy. Report of a case]. AB - The authors report a case, the first in Morocco, of a kidney transplanted woman pregnancy. The pregnancy occurred eleven months after the kidney transplantation. The renal function has not been modified. Treatment immunosuppressor has been maintained. The pregnancy evolution has been marked by a moderate hypertension arterial without child reverberation. To shortcoming this observation and to the lights of data of the literature, authors recalls factors that condition the stability of the renal function and the good progress of pregnancy and insist on the necessity of the scheduling of pregnancy and a multidisciplinary surveillance. PMID- 11496593 TI - [Verrucous carcinoma of the kidney: report of 2 cases]. AB - The verrucous carcinoma is an unusual shape of well differentiated squamous cell carcinoma, first described at the ORL region; the kidney location is rare; the risk factors are represented essentially by lithiasis and/or urinary infection; the clinical symptom is not specific. Diagnosis is facilitated by radiological investigations and particularly excretory urogram/ultrasound; certainly diagnosis is pathological. The nephro-ureterectomy with collar resection of the bladder is the choice treatment. We report two observations and we clarify clinicopathological aspects of this type of carcinoma and we discuss the prognosis. PMID- 11496594 TI - [Percutaneous nephrostomy: use of a simple inexpensive material in emergencies in precarious conditions]. AB - We describe a cheap and simply technique of percutaneous nephrostomy which can be used anywhere. It consists, under local anesthesia, of insertion of a drain into renal cavity using only a needle, a sheat and dilatator. With this technique, we placed 81 PCN with 97% of success, without complication. PMID- 11496595 TI - [Treatment of ureteral calculi with rigid ureteroscopy. Report of 67 cases]. AB - PURPOSE: To evaluate the results obtained by rigid ureteroscopy to treat ureteral calculi. MATERIALS AND METHODS: Between May 1998 and September 2000, 63 patients underwent 67 rigid ureteroscopies (URS) with three endoscopic retreatments and one bilateral URS, for 48 distal, ten mid and nine proximal ureteral calculi. RESULTS: After URS 77.6% of the patients were free of the stones. Success rate was respectively 85.4%, 60% and 55.5% in distal, mid and proximal uretero. Morbidity was 10.4% and there have been one stricture treated by endoscopy. CONCLUSION: Our series demonstrates that rigid URS is safe and effective procedure to treat distal, ureteral calculi. Thus, it should be considered a treatment of choice for distal ureteral calculi. PMID- 11496596 TI - [Urologic complications of gynecologic surgery. Report of 16 cases]. AB - From 16 cases collected in eight years (January 1991-December 1999) in the I. Deen Teaching Hospital Department of Urology, the authors study the gynaecological and obstetric surgery urological complications. They noted that these complications interest in 80% of cases the 18-47 years old woman with a high parity in 70% of cases. Hysterectomy, whatever the indication is the main etiology: 62.50%, caeserean takes the second place with 25%. The anatomic lesions are ureteral in 68.75% of cases and bladder in 31.25%. In the 14 cases treated, in the department, uretero reimplantation has been done in 46.66%, bladder suture in 40% and nephrectomy in 6.66%. Thirteen women were healed and one died. PMID- 11496597 TI - [Hydatid cyst of the Douglas' cul-de-sac with fistula to the bladder. Report of 2 cases]. AB - The present study reported two cases of hydatid cysts localized in Douglas space and that broke in the bladder. The clinical course was characterized by urinary symptoms and by presence the cysts in urine. Abdominopelvic ultrasonography is essential for diagnosis and identification of other sites. When diagnosis is unclear, then CT scan plays a role. Therapy included cystectomy and pericystic resection in one case and intravescical injection of H2O2 in the second case. The course was favorable in both cases. PMID- 11496598 TI - [Foreign bodies of the bladder. Report of a new case]. AB - In this paper we are relating the medical history of 30 year-old female patient with psychiatric medical background. This patient introduced multiple foreign bodies in her bladder via the urethra. Throughout this observation, we analyse diagnosis and therapeutic aspects of bladder foreign bodies. PMID- 11496599 TI - [Comparison of 3 self lubrificated urethral catheters: prospective study on 27 patients]. AB - Clean intermittent catheterization has become a usual procedure in urologic practice. Self lubrificated catheters are nowadays accepted as the most efficient. This randomised and prospective study compared tolerance and efficiency in a group of 27 patients of three well known catheters (12 charriere, straight): Flocath (Ruschcare), Lofric (Astratech) and Easycath (Coloplast). The systematic comparison of this three catheters did not allow to put in a prominent position one of these. The three catheters are extremely close in terms of tolerance and performance. PMID- 11496600 TI - [Perineoscrotal gangrene: report of 51 cases. Diagnostic and therapeutic features]. AB - OBJECTIVE: To approach the treatment of Fournier's gangrene. So, pathophysiology and etiology are recalled. METHODS: The authors present 51 cases of Fournier's gangrene treated from 1989 to 1998, their age ranged from 19 to 89 years. Data were collected on admission signs and symptoms, physical examination. Aggressive surgical debridement of all necrotic tissues was performed, Intravenous antibiotics and resuscitation fluid were also administered. RESULTS: All patients were male. In 20 cases (39%), there was no identifiable cause, and in 31 cases (61%), the etiology of gangrene was urethral (33%), anorectal (28%) and unknown (19%). The average hospital stay was 30 days. Three cases underwent unilateral orchidectomy, six colostomy and in 17 cases, a suprapubic catheter was inserted. Mortality was high (18%) and essentially associated to debilated state and toxi infectious context. CONCLUSION: Fournier's gangrene is a true urologic emergency potential lethal, which requires aggressive antibiotic and surgical treatment. PMID- 11496601 TI - [Testicular metastasis of prostatic adenocarcinoma. Report of a case]. AB - Metastasis involvement of the testis form prostatic carcinoma is rare. Seventy cases have been reported between 1938 and 1990. We report another case treated by transurethral resection of the prostate, bilateral orchiectomy and nonsteroid androgens and a review of the literature is presented. PMID- 11496602 TI - [Xanthogranulomatous orchitis. Report of 7 cases]. AB - The xanthogranulomatosis orchiepididymititis is a benign and a rare disease. Its etiology is unknown. It creates diagnosis, therapeutic and etiological problems. The diagnosis is hardly made because of the variability of the clinical findings. The confusion is mainly made with a malignant tumor, and sometimes with a simple orchiepididymitis. Orchidectomy is usually performed. We report seven cases of orchiepididymitis characterized by their antibiotherapeutic resistance, the destruction of the testis which needed a surgical removal. The diagnosis is established after the histological study of the testis. The etiology is not yet known. The authors study the clinical evolution and therapeutic characteristics of our serial comparing with the literature data. PMID- 11496603 TI - [Abdominoscrotal hydrocele]. AB - The abdominoscrotal hydrocele (ASH) is a rare variant of hydrocele of the tunica vaginalis with fluid accumulation into the abdominal cavity passing through the inguinal canal. The etiology of ASH is not clear. The positive diagnosis depends on the clinical data (hydrocele + abdominal mass). It is confirmed by means of echography and or by TDM. The treatment is surgical complete resection of the vaginalis with ligation of the peritoneal cavity. This report concerns the observation of a huge ASH, so far described in a literature review. We try to distinguish the diagnostic and therapeutic elements of this exceptional pathology. PMID- 11496604 TI - [The hand in the Nile's course]. PMID- 11496605 TI - [Treatment of recurrent Dupuytren's disease. 1992]. PMID- 11496606 TI - [Percutaneous needle aponeurotomy. Complications and results]. AB - Recently French rheumatologists have repopularized fasciotomy using a percutaneous needle technique. This blind approach has been claimed to be plagued by numerous complications. We reviewed the charts of 211 patients treated consecutively on 261 hands and 311 fingers to assess the rate of postoperative complications. The first 100 patients were evaluated with a mean follow up of 3.2 years to assess the rate of recurrences and extension of the disease. In the whole group the mean age was 65 years and delay between onset and treatment was six years. Rupture of the cords were performed only in the palm in 165 cases, in the palm and finger in 111 and purely at digital level in 35. Complications were scarce without infection or tendon injury but one digital nerve was found injured during the second procedure. Postoperative gain was prominent at metacarpophalangeal joint level (79% versus 65% at interphalangeal level). Reoperation rate was 24%. In the group controlled at 3.2 years follow up, recurrence rate was 58% and disease "activity" 69%. Fifty nine hands need further surgery. The ideal indication for this simple and reliable technique is an elderly patient with a bowing cord and a predominant MP contracture. PMID- 11496607 TI - [Isolated fifth digit localization in Dupuytren's disease]. AB - Isolated fifth digit localisation in Dupuytren's disease has a bad reputation. A series of this injury is reported with a special attention on recurrences. This series is composed of 30 cases in 26 patients (four bilateral cases): the majority of patients were male and 53.5 y was the average age. Extension loss of MP and PIP joints were present in 25 cases, MP isolated extension loss in two cases and PIP in three cases. The treatment performed was a percutaneous needle technique in five cases, surgical fasciectomy in 24 cases using a zigzag palmodigital longitudinal fasciectomy approach in 18 cases, an open palm technique in six cases and one isolated laterodigital flap. Assessment of correction was based on Tubiana's classification. Average follow-up was 22 months. Postoperative course was uneventful except for one case of precarious vascularisation which leads to an amputation. For the digitopalmar localisations: 15 stage 0 or 1, 7 stage 2, 1 stage 3 and 1 stage 4. Improvement percentage was 0.60 in combined cases, 0.65 in cases with a MP loss and 0.46 in PIP loss. Five recurrences were noted. Literature on this topic is coherent with the fact that isolated involvement of the fifth digit carry a bad prognosis, mainly due to the high recurrence rate in our series as in other papers. In severe injuries, an imperfect result should be the aim to prevent vasculonervous complications. PMID- 11496608 TI - [Unstable fracture of the distal radius and its treatment: comparison of three techniques: external fixation, intramedullary pinning and AO plates]. AB - This retrospective study compares 94 distal radius fractures. Sixty one women (median age 66) and 33 men (median age 42) were reviewed clinically and radiologically by a surgeon not involved in treatment. Fractures were classified into three AO groups. Standard X-rays were used for radiological evaluation. Fractures of the distal ulna were evaluated separately. Chi-square tests, Wilcoxon, Mann-Whitney and Fisher's tests were used for statistical evaluation. Women presented mainly "A" type fractures. They were treated by intramedullary pinning. Forty two fractures were grafted, using autogenous bone for young patients and bone substitute for the elderly. Twelve scapholunate dissociations were recorded, all but one were detected in women, 50 years of age or more. Four were painfree, two presented climatic pain and six claimed pain during effort; none had snapping. Radio-ulnar laxity was similar with or without styloid fractures. External fixator and autogenous grafts appeared the most efficient technique for maintaining radial length. Ulnar head fractures were related to a significant higher incidence of sympathetic dystrophy with reduced prono supination, a correlation not previously noted to your knowledge. This suggests that radius and ulnar head fracture should be classified independently. The amount of pain was not related to classification, internal fixation or gender. PMID- 11496609 TI - [Orthopedic treatment of fifth metacarpal neck fractures: prospective study]. AB - Many methods of treatment have been proposed for the metacarpal neck fractures of the little finger, from early mobilisation to open or closed surgical techniques. A prospective studies of 20 cases treated by a non surgical technique is presented. The reduction was performed by manipulation under local anesthesia according to the Jahss technique and a modified Thomine brace was applied for four weeks. All patients were reviewed for follow-up at 30, 60, and 180 days. The fracture displacement was initially 34 degrees. It was reduced by manipulation to 12 degrees. All fractures healed with an average final displacement of 20 degrees. At 6 months for final examination, average T.A.M. was 245 degrees, T.P.M. was 276 degrees, grip strength reached 96% of the other hand. There were 19 excellent and one bad results. In conclusion, ambulatory non surgical treatment by reduction and local immobilisation of the fractures of the fifth metacarpal provide a good final result with a low cost. PMID- 11496610 TI - [Tension force in dynamic splints made of neoprene]. AB - Dynamic splints are the most difficult ortheses to make. Unrestricted finger joint mobility is usually prevented by the size of their components. Ortheses made out of neoprene material have the advantage of being pliable and at the same time can be constructed as dynamic splints. Neoprene material elasticity allows conforming it into a tube that can be fitted over a finger, spreading uniform pressure. A traction slip can be cemented to the neoprene finger tube. With neoprene splints, lever arms are more efficient. Their tubular design applies and spreads traction and countertraction forces in all directions. Neoprene material's elasticity contributes in reducing edema formation as well as distributes forces applied to the finger over a large area. The risk of excessive pressure is the only drawback of an elastic tubular design, it may cause pain and tissue ischemia. A meticulous tailoring prevents this potential risk. PMID- 11496611 TI - [Giant cell tenosynovial tumor, spread in the forearm soft tissues. A case report]. AB - Giant cell tumor of the tendon sheath (GCTTS), tenosynovial giant cell tumor are a group of rare proliferative disorders which involves synovial joints and tendon sheaths. Considerable confusion exists about the surgical treatment and diagnosis of these disorders. We report an exceptional case of a tenosynovial giant cell tumor of the forearm. It concerns a 23 years old woman presenting multiple tumors of the forearm and wrist. Result of the primary biopsy concluded to an eosinophilic granuloma. Magnetic resonance imaging demonstrated a richly vascularized tissular lesion involving the extensor carpi radialis and extensor pollicis longus. Surgical excision of the tumor was performed. Macroscopic, histologic and immunohistochimic findings confirmed the diagnosis of tenosynovial giant cell tumor. This diffused form is rare compared with the nodular one involving fingers. Optimal treatment is surgical resection. The only risk is recurrence. PMID- 11496612 TI - [Compression of the anterior interosseous nerve by Gantzer's muscle]. AB - The authors report a case of incomplete anterior interosseous nerve syndrome due to mechanical compression by Gantzer's muscle. This accessory muscle induced an isolated paralysis of the flexor pollicis longus with a characteristic pinch attitude. No improvement was shown after six weeks of medical treatment. Recovery was rapid and complete following surgical relief of compression. Anatomical considerations, clinical features, and treatment are discussed. PMID- 11496614 TI - [What importance have original papers on the prevention of nausea and vomiting in our clinical practice?]. PMID- 11496613 TI - Bizarre parosteal osteochondromatous proliferation of the hand: report of two cases. AB - Bizarre parosteal osteochondromatous proliferation is a rare tumor usually localized in the hand. We report two new cases, one in the thumb of a middle aged woman and one in the terminal phalanx of a young haemophiliac patient. PMID- 11496615 TI - [Natriuretic peptides: physiological, pathophysiological and clinical aspects]. AB - A milestone was reached in cardiophysiology when in 1981 DeBold demonstrated that the heart functions as an endocrine gland by injecting an extract of atrial muscle into rats, resulting in an induction of natriuresis and a drop in blood pressure. This observation then led to the discovery of a family of related peptides with slightly different amino acid compositions working in concert to achieve the maintenance of sodium and volume homeostasis. The natriuretic peptide family consists of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), and Urodilatin (URO) with their tissue specific distribution including the heart (ANP, BNP), brain (ANP, BNP, CNP), endothelial cells (CNP), and kidney (URO). These peptides were thought to be primarily involved in cardiovascular and renal functions but have now proven to play a role in other physiological systems. In view of their known biological effects, therapeutic efficacy from administration of ANP, BNP or URO might be anticipated, for example in acute renal failure or congestive heart failure. A number of clinical trials suggest that application of these peptides may represent a new pharmacological tool in the treatment or prevention of these diseases, but the clinical benefit still needs to be shown in large controlled studies. In addition to therapeutic options it is possible that plasma concentrations of ANP and BNP could play a role as diagnostic and prognostic markers of cardiac dysfunction. PMID- 11496618 TI - [Anesthesia induction by face mask in children: pro]. PMID- 11496616 TI - [Analgesic dosage with (S)-ketamine/propofol vs. (S)-ketamine/midazolam: sedation, stress response and hemodynamics--a controlled study of surgical intensive care patients]. AB - OBJECTIVE: The study was undertaken to investigate the influence of two different regimens of analgosedation on control and quality of sedation, stress response and haemodynamic parameters. METHODS: After ethical approval, 30 surgical intensive care patients were investigated in an open, controlled design. Patients with initial cardiocirculatory stability received 0.33-1.0 mg/kg BW/h (S) ketamine together with 1-3 mg/kg BW/h propofol (SK/P-group), whereas patients with impaired cardiocirculatory stability received 0.33-1.0 mg/kg BW/h (S) ketamine and 0.033-0.1 mg/kg BW/h midazolam (SK/M-group). Analgosedation was titrated until tolerance of respirator treatment was achieved and the patient was asleep, but able to respond to simple commands. At least 12 h after beginning of analgosedation, a simple neurological examination ("diagnostic window") was undertaken. RESULTS: In both groups, biometric data and diseases were altogether comparable, and tolerance of respirator treatment was excellent. About 16 h after start of analgosedation, 13 of 14 patients (93%) in the SK/P-group were immediately cooperative. In 2 of 16 patients of the SK/M-group, self extubation occurred, and 9 of 14 remaining patients (64%) were immediately cooperative (p = 0.065). Assessment of control and quality of analgosedation indicated slight advantages in SK/P-patients. SEF90 showed predominant beta-activity in both collectives, which increased in the course of time. Adrenaline, noradrenaline, ADH, ACTH and cortisol were measured at 7 time points. All endocrine stress parameters were consistently above normal range, but decreased during the observation period (p < 0.05). In the SK/M-group, ADH was significantly and noradrenaline initially higher than in controls. Systolic arterial pressure was comparable, whereas heart rate was significantly lower in the SK/P-group (p = 0.001). No relevant changes of endocrine or haemodynamic parameters were observed at neurological examination. CONCLUSION: In surgical intensive care patients, analgosedation with SK/P showed some advantages over SK/M with respect to control and quality. The endocrine stress response was reduced by both regimens in course of time. Altogether higher levels of ADH and noradrenaline during SK/M analgosedation let expect higher cardiocirculatory stability and possible reduction of catecholamine demand. Due to ketamine-typical beta-activity, a reliable assessment of sedation by the pEEG is not possible. PMID- 11496617 TI - [Prophylaxis of nausea and vomiting after thyroid surgery: comparison of oral and intravenous dolasetron with intravenous droperidol and placebo]. AB - OBJECTIVE: Postoperative nausea and vomiting (PONV) are among the most common complications in operative medicine. Especially thyroid surgery is frequently associated with PONV. It was the aim of this study to determine the efficacy of oral and intravenous dolasetron in comparison to intravenous droperidol (DHB) and placebo in the prevention of PONV. METHODS: 93 female and 43 male patients undergoing thyroid surgery were stratified according to gender and then randomised to receive double-blind one of four antiemetic regimes: 50 mg dolasetron given orally 45 minutes prior to induction of anaesthesia (group I), 12.5 mg dolasetron given intravenously during induction of anaesthesia (group II), 1.25 mg DHB given intravenously during induction of anaesthesia (group III) or placebo (group IV). General anaesthesia and preoperative management of the patients were standardised: premedication with chloracepate-dipotassium, induction with thiopentone, sufentanil and rocuronium, maintenance with N2O/O2, sevoflurane and repetitive doses of sufentanil and rocuronium, postoperative analgesia with metamizol and piritramide, antiemetic rescue-treatment with dimenhydrinate, metoclopramide and triflupromazine. Number of emetic episodes, the need for additional antiemetics and adverse events were recorded for 24 hours. Efficacy was measured by "complete-response" (CR = 0 emetic episodes or 1 emetic episode after 4 hours and no rescue-treatment) and "total-response" (TR = complete response plus no nausea, i.e., < 5 mm VAS rating of patients maximum nausea). RESULTS: Men: Only Dolasetron given intravenously reduced nausea and vomiting significantly, Dolasetron given orally reduced nausea, but not vomiting, DHB had no significant effects: CR 72.7% (group I), 100% (group II), 80% (group III), 63.6% (group IV); TR 72.7% (group I), 81.8% (group II), 50% (group III), 36.4% (group IV). Women: In all three treatment groups significantly less patients suffered from PONV compared to the placebo group (p < 0.05). There were no differences between the treatment groups: CR 58.3% (group I), 45.8% (group II), 52.2% (group III), 18.1% (group IV); TR 37.5% (group I), 33.3% (group II), 39.1% (group III), 13.6% (group IV). There were no adverse events in any group. CONCLUSIONS: Our results confirm the expected high incidence of PONV after thyroid surgery, especially in female patients. Single doses of oral and intravenous dolasetron and intravenous droperidol reduced PONV effectively in female patients undergoing thyroid surgery. Dolasetron seems to be the more effective substance in male patients. Both substances can be administered safely and are well tolerated. PMID- 11496619 TI - [Anesthesia induction by face mask in children: contra]. PMID- 11496620 TI - [Anesthesia in mitochondrial encephalomyopathies]. AB - A 6-year-old boy with a rare mitochondrial disease (MELAS: mitochondrial encephalopathy, lactic acidosis, stroke-like episodes) was presented to undergo adenoid resection and bilateral paracentesis. ENT surgery was performed without complications under general anaesthesia using propofol, fentanyl, and ventilation with nitrous oxide and oxygen. Routine intraoperative monitoring (ECG, noninvasive blood pressure, oxymetry and capnometry) was supplemented by frequent body temperature measurements and repeated laboratory analysis of venous blood gases, lactate, and glucose. Clinically, the postoperative course was uneventful and the boy was discharged from hospital on the first postoperative day. Signs or symptoms of malignant hyperthermia never occurred. Laboratory analysis only showed a remarkable serum lactate elevation postoperatively (6 mmol/l) which decreased on the first postoperative day (3.7 mmol/l). The present anaesthesiologic experiences with MELAS-syndrome are limited, and recommendations are mainly based on case reports. Careful preoperative physical examination with special regard to all available medical records, and anaesthetic management comparable with that in malignant hyperthermia susceptible resulted in an uneventful course in our patient. Pathogenetic aspects of mitochondrial diseases focussing on anaesthetic considerations are briefly discussed. PMID- 11496621 TI - [Genetically engineered drugs and their application with the example of erythropoietin]. AB - Over the past two decades, many genetically engineered drugs have been developed and approved for the treatment of patients. Typically, these drugs are characterized by a high and specific activity in the presence of optimal safety. They include hormones, enzymes, growth and coagulation factors, antibodies as well as vaccines. All these proteins are generated using recombinant DNA technology. An expression vector with the gene encoding for the protein of interest is introduced into an appropriate microorganism or cell line. The biochemical machinery of the host cell then translates the genetic information into the corresponding protein. Large scale production of the recombinant drugs uses biotechnological processes. The genetically modified organisms are grown in bioreactors from which the desired protein is finally isolated and purified. This review focuses on the production and clinical application of recombinant erythropoietin in the areas of nephrology, hemato-oncology and elective surgery. PMID- 11496622 TI - Research as a foundation for action on gambling. PMID- 11496623 TI - Updating and refining prevalence estimates of disordered gambling behaviour in the United States and Canada. AB - BACKGROUND: This study updates prevalence estimates of gambling-related disorders in the United States and Canada, identifies differences in prevalence estimates among population segments, and identifies changes in prevalence over the past 25 years. METHOD: A meta-analytic strategy guided the synthesis of 180 estimates derived from 146 prevalence studies. RESULTS: Prevalence estimates among adolescent samples were significantly higher than estimates among adult samples for both clinical (level 3) and sub-clinical (level 2) measures of disordered gambling within both lifetime and past-year time frames. Among adults, level 3 prevalence estimates continue to increase significantly. CONCLUSIONS: Membership in youth, treatment, and prison population segments is significantly associated with experiencing gambling-related disorders. Understanding sub-clinical gamblers provides a meaningful opportunity to lower the public health burden associated with gambling disorders. Prospective studies of incidence are necessary to determine whether the prevalence of disordered gambling continues to increase among the adult general population and how adolescent gambling experiences change as this cohort ages. PMID- 11496624 TI - Comparing benefits and risks of immunization. AB - BACKGROUND: This paper uses a straightforward method of quantifying the benefits and risks of immunization, illustrated by universal immunization programs in British Columbia against invasive Haemophilus influenzae type b disease (Hib), measles, rubella, and paralytic poliomyelitis. METHODS: Data were extracted from provincial communicable disease and vaccine adverse event surveillance systems. Average disease incidence was compared before and after implementing universal immunization programs. Estimates of prevented deaths and serious disease complications were calculated and compared with expected numbers of serious vaccine-associated adverse events (VAAEs). RESULTS: Average incidence of reported cases decreased 90 to 100% over a 5-year period after implementing universal programs. These benefits were sustained or strengthened over time. Rates of reported serious VAAEs were low. DISCUSSION: The remarkable success of immunization has created a paradox. Despite a low absolute risk of serious VAAEs, the relative risk of some VAAEs can exceed risk of disease in the province. PMID- 11496625 TI - The role of the health sector in addressing poverty. AB - PURPOSE: To explore Canadian health sector initiatives addressing poverty. METHODS: Information about 224 health sector initiatives addressing poverty was collected from Health Canada, provincial/territorial health ministries, and health regions. RESULTS: Health Canada, 12 provincial/territorial health ministries, and at least one third of health regions have been undertaking poverty-related initiatives. Almost two thirds (64.7%) of initiatives focused on the consequences of poverty. Much less frequent were initiatives that aim to: raise awareness about poverty; prevent people from becoming poor; enhance skills and education of people in poverty; and alter social and economic conditions contributing to poverty. DISCUSSION AND CONCLUSIONS: While strategies that focus on the consequences of poverty likely enhance the health of Canadians in poverty, these strategies do little to reduce poverty rates. Efforts to improve the health of both individual Canadians in poverty and society as a whole will be limited until the health sector uses more strategies that challenge fundamental structural conditions contributing to poverty. PMID- 11496626 TI - Do area-based markers of poverty accurately measure personal poverty? AB - Area-based markers of deprivation (e.g., postal codes) are commonly used to identify groups of people with low socioeconomic status (SES); the validity of this approach, however, remains unknown. In this study, we determined the accuracy of using income quintile groups calculated on the basis of the median family income of each forward sortation area (1996 Canadian census) to identify those living in poverty (i.e., annual family income of < $12,620). The sensitivity and specificity of using the lowest income quintile to capture those in poverty were 26% and 83%, respectively (likelihood ratio (LR) of 1.49; 99% CI, 1.49 to 1.50). Among those in non-metropolitan and metropolitan areas, the LRs were 1.26 (99% CI, 1.26 to 1.27) and 2.01 (99% CI, 2.01 to 2.02), respectively. The use of postal codes as the only marker to identify people with low SES may result in substantial misclassification of personal poverty, particularly in non metropolitan areas. PMID- 11496627 TI - Predictors of Canadian legislators' support for public health policy interventions. PMID- 11496628 TI - How government policy decisions affect seniors' quality of life: findings from a participatory policy study carried out in Toronto, Canada. AB - Toronto seniors explored how government policy decisions were influencing their health and well-being. In this participatory policy study, emphasis was upon the lay and critical knowledge of highly informed seniors. Focus groups and interviews revealed that all three levels of governments were seen as not listening to seniors' voices. In nine key policy areas identified as influencing seniors' quality of life--hearing seniors' voices, housing, acute illness care, long-term care, income supports, transportation and mobility, promoting healthy lifestyles, access to information, and hearing voices from cultural communities- many concerns were raised. The gap between government rhetoric and government action on seniors' issues merits public health attention. PMID- 11496629 TI - Understanding newborn infant readmission: findings of the Ontario Mother and Infant Survey. AB - The Ontario Mother and Infant Survey examined health and social service utilization of postpartum women and newborn infants from five hospital sites. A cross-sectional multilanguage survey design with longitudinal follow-up was used: 1,250 eligible, consenting women completed a self-report questionnaire in hospital and 875 women participated in a structured telephone interview at four weeks post-discharge. Rates of newborn infant readmission ranged from 2.4% to 6.7%. The best predictors of readmission were: main source of household income was other than employment; maternal self-rating of health was poor; mother anticipated inadequate help and support at home following discharge; mother received help from friends/neighbours following discharge; and mother had concern about infant care and behaviour. Readmission was not associated with length of postpartum hospital stay. The study findings suggest that there is a complex relationship between infant health care needs, family resources and provider practices that produces clinically important, site-specific readmission patterns. PMID- 11496630 TI - Injury in Canadian youth: a brief report from the Health Behaviour in School-Aged Children Survey. PMID- 11496631 TI - Harm reduction and illegal drugs: the real debate. PMID- 11496633 TI - General social support and physical activity: an analysis of the Ontario Health Survey. AB - In spite of the many benefits of regular physical activity, the majority of Canadians are sedentary. This paper examines the relationship between general social support and physical activity levels. An analysis of data on 29,135 individuals from the 1990 Ontario Health Survey was conducted. Hierarchical multiple regression was used to determine the relationship between measures of general social support, derived from factor analysis, and physical activity levels. Sociodemographic and perceived health status measures were included as control variables. The results showed that general social support, in the form of Social Quantity and Social Frequency (number of friends/family members and frequency of contact), was significantly associated with higher levels of physical activity. Conversely, support in the form of Familial Structure (marital/cohabitation and parental status) was significantly associated with lower levels of physical activity. The findings indicate the types of general social support that facilitate or hinder participation in physical activity. PMID- 11496632 TI - Familial influences on substance use by adolescents and young adults. AB - This study uses data from the Ontario Health Survey to examine within-family influences (sibship number, age and sex composition; family structure and parental substance use) on the use of tobacco, alcohol and marijuana in households (N = 4,643) among offspring aged 12 to 24 years. Using a modification of the kappa statistic, concordance among siblings is modest generally and undifferentiated across substance type. Concordance is stronger among sibships that are either all male or older (19-24 years) and is particularly strong for siblings < or = two years apart in age. The dominant influence of substance use behaviour appears to be from older siblings to younger siblings and not from parents to offspring. Sibling concordance for substance use suggests that the treatment and prevention of substance use (and abuse) among adolescents and young adults might be enhanced by including a family focus, especially where there are two or more siblings at home. PMID- 11496634 TI - The effects of physically active leisure on stress-health relationships. AB - In this article, the effects of physically active leisure on the relationships between stress and health are examined using structural equation modeling (SEM). The analyses are based on data from Canada's 1994 National Population Health Survey (n = 17,626). Overall, physically active leisure was found to directly contribute to higher levels of physical health and wellbeing, and lower levels of mental ill-health among Canadians. When the respondents experienced higher levels of chronic stress, life event stress, and/or work stress, involvement in physically active leisure appeared to help them maintain good health and wellbeing. Also, higher levels of participation in physically active leisure helped paid workers suppress levels of work stress. Agencies involved in health promotion and lifestyle intervention should give greater consideration to physically active leisure. As a significant component of an active lifestyle, physically active leisure can contribute to better health, and provide a valuable resource for coping with stress. PMID- 11496635 TI - Nova Scotia high school students' interactions with physicians for sexual health information and services. AB - PURPOSE: To support a community effort to establish a health service in a Nova Scotia high school, adolescents' sexual behaviours and use of physicians for sexual health services were assessed. METHODS: A self-completion survey asked students about sexual behaviours, use of physician services and barriers to use of those services. RESULTS: Only 8.7% of male and 37.9% of female students with family physicians had discussed with the physician whether they were sexually experienced (p < 0.0001). More females had discussed sexual activity when the physician was female than when the physician was male (43.7% vs. 35.1%; p < 0.05). More sexually experienced females who had not discussed this behaviour with their physicians identified specific issues as barriers to discussion. CONCLUSIONS: Female physicians discuss sexual activity more with their adolescent female patients than do male physicians. The results support the need to provide health services in addition to those of physicians to meet the sexual health needs of adolescents. PMID- 11496636 TI - [Evaluation of mass screening activity for cardiovascular disease risk factors in a rural environment]. AB - The aim of this study was to evaluate the immediate and short-term impacts of a cardiovascular mass screening activity in a rural environment. A posttest-only control group design was used. Concerning the immediate impact, the results showed that the participants of the experimental group had, compared to the participants in the control group, a higher intention to participate regularly in physical activities and a higher intention to reduce their fat consumption. The participants' intention to stop smoking was not modified by the screening activity. After four months, the results revealed an increase in participation in physical activities and a decrease in fat consumption. The proportion of participants who smoked did not decrease. In conclusion, it is suggested that a cardiovascular mass screening activity such as the one performed in this study could be effective in motivating a population to adopt heart health-related behaviours. Others studies are necessary however to confirm the short-term impact of this type of activity on behaviour modification with regard to cardiovascular health. PMID- 11496638 TI - Health informatics education: an opportunity for public health in Canada. AB - Health information infrastructure is being developed across Canada, and health informatics education should be a component of the emerging infrastructure. However, educational opportunities do not appear to be developing in pace with infrastructure. This study characterizes the required education, and describes specific issues facing the development of health informatics education. Twenty six key informants were interviewed, and responses were analyzed to identify major themes. Subjects agreed that the current capacity for health informatics education is inadequate. Also, limited expertise could adversely affect health infostructure implementation and health system performance. A need was identified for both advanced and basic education. Four issues facing the development of health informatics education in Canada were consistently identified: awareness, collaboration, funding, and human resources. Public health is well positioned to play a central role in health informatics education due to its historical basis in handling health information, and its developing expertise in health informatics applications. PMID- 11496637 TI - [1998 Quebec Social and Health Survey: determinants of chronic respiratory diseases]. AB - In the course of the "1998 Health and Social Survey", questions were included to verify the prevalence of chronic respiratory diseases and also of wheezing. The objectives of this study were 1) to verify the prevalence of wheezing and its validity as an indicator of chronic respiratory diseases in Quebec; and 2) to examine the relationship between chronic respiratory diseases and some of their potential determinants. A total of 30,386 individuals participated in the study. For all ages, the prevalence of wheezing was 5.4%. It was associated with asthma, allergies, chronic bronchitis and emphysema. A low familial income and tobacco smoking were associated with wheezing, asthma, chronic bronchitis and emphysema. Passive smoking was associated with wheezing whereas the presence of carpets was associated with wheezing and asthma. Between 32 and 48% of families with an asthmatic or an allergic member modified their dwelling to alleviate respiratory problems. The prevalence of wheezing documented here was lower than in anglosaxon countries. This result could be explained by a cultural factor (the French translation or the perception of wheezing). This study emphasizes the role of reducing tobacco smoking in the prevention of chronic respiratory diseases. PMID- 11496639 TI - Re: Ostry A. The new international trade regime: problems for publicly funded healthcare in Canada? (Editorial) CJPH 2001; 92(1):5-6. PMID- 11496640 TI - [Experience with the diagnosis and treatment of arthropathy caused by hemophilia]. PMID- 11496641 TI - [Digestive system complications of idiopathic hypoparathyroidism]. PMID- 11496642 TI - [Clinical picture of arthritis caused by psoriasis]. PMID- 11496643 TI - [Cranio-caudal axial incidence in the radiologic examination of sacro-iliac articulation]. PMID- 11496644 TI - [Ankle aspects of amyloidosis]. PMID- 11496645 TI - [Levamisole used in rheumatoid arthritis]. PMID- 11496646 TI - [Hematologic side effects of levamisole in the treatment of rheumatoid arthritis]. PMID- 11496647 TI - [Clinical aspect of bacterial infections of peripheral joints and spinal column]. PMID- 11496648 TI - B27 and agranulocytosis in rheumatoid arthritis patients treated with levamisole. PMID- 11496649 TI - [Study of a population of HLA-B27-positive blood donors]. PMID- 11496650 TI - Immunogenetic implications in rheumatic diseases. PMID- 11496651 TI - [Study of the phenomenon of discordance of HLA-B27 during the course of ankylosing spondylitis]. PMID- 11496652 TI - HLA B27 prevalence in spondylo-arthropathy syndromes. PMID- 11496653 TI - Acute anterior uveitis. A clinical, HLA and scintiscan survey. PMID- 11496654 TI - [HLA-B27 antigen and seronegative arthritis]. PMID- 11496655 TI - Rheumatic disorders in a HLA B27 positive population. PMID- 11496657 TI - HLA antigens in rheumatic diseases. Introduction. PMID- 11496656 TI - HLA typing in clinical rheumatology. PMID- 11496658 TI - [HLA antigens and Heberden nodes]. PMID- 11496659 TI - EU policy on nutrients emissions: legislation and implementation. AB - After 25 years of EU water legislation the European Union has just thoroughly restructured its water policy. The European Parliament and the Council, following a tough conciliation procedure between the two legislators, have in summer 2000 agreed a proposal by the European Commission for a Water Framework Directive. This legislation will have the following main objectives: integrated river basin management across borders, with coordinated programmes of measures protection of all waters, surface waters and groundwater, in quality and quantity with a proper ecological dimension emissions and discharges controlled by a "combined approach" of emission limit values and quality standards, plus the phasing out of particularly hazardous substances introducing water pricing policies strengthening public participation This new Water Framework Directive adopted in September 2000 will complement existing EU water legislation on nutrients reduction--the 1991 Directive on nitrates pollution from agricultural sources and the 1991 Directive on urban waste water treatment. These Directives will remain main pillars of EU water policy whilst at the same time being integrated into the river basin management in a coherent way. PMID- 11496660 TI - Innovative low cost procedure for nutrient removal as an integrated element of a decentralised water management concept for rural areas. AB - The absence of large rivers with rather high niveau of self purifying effect in parts of east Germany leads to a discharging of the effluent of wastewater treatment plants into the groundwater in many cases. One useful consequence is the idea of realisation of decentralised measures and concepts in urban water resources management concerning municipal wastewater as well as rainfall, precipitation. At the same time, only the upper soil zone--a few decimetres--is water--saturated and thus discharge effective, even when extreme rainfall takes place. Underneath, however, there generally exists an unsaturated soil zone, which is up to now a rather unexplored retardation element of the hydrologic- and substrate-cycle. Nutrient removal in small wastewater treatment plants that are emptying into ground waters is often beneficial. The presented studies optimised an inexpensive method of subsequent enhanced wastewater treatment. The developed reactor is similar to a concentrated subsoil passage. The fixed bed reactor is divided in two sections to achieve aerobic and anoxic conditions for nitrification/denitrification processes. To enhance phosphorus removal, ferrous particles are put into the aerobic zone. Two series of column tests were carried out and a technical pilot plant was built to verify the efficiency of the process. The results show that this method can be implemented successfully. PMID- 11496661 TI - Influence of primary sedimentation on pre-denitrification system performances. AB - The effects of suppressing primary sedimentation on nitrogen removal efficiency of a pre-denitrification system have been evaluated for a large municipal wastewater treatment plant. Simulations have been carried out using the STOAT model. For both the process schemes with and without primary sedimentation, nitrification efficiencies are calculated for increasing influent loads of COD, total N and suspended solids. The sensitivity analysis shows that for the usual carbon to nitrogen ratios in the raw influent both the process schemes allow the requested removal efficiencies, whereas for significantly high C/N ratios the scheme with primary sedimentation is preferable. PMID- 11496662 TI - Retrofitting SBR systems to nutrient removal in sensitive tourist areas. AB - Retrofitting of existing SBR systems for nutrient removal is evaluated and defined for small communities in sensitive coastal areas, with seasonal fluctuations in wastewater quantity and quality. The proposed approach is developed by means of basic process stoichiometry and verified using ASM2d. The efficiency of retrofitting is found to rely on the delicate balance between the overall sludge age, the initial settled sludge volume in the reactor, and the ratio of the initial volume to the feed volume in each cycle, a parameter corresponding to the recycle ratio in continuous systems. PMID- 11496663 TI - What to do after nutrient removal? AB - In the Netherlands, interest in advanced treatment is increasing now that almost all wastewater treatment plants apply full biological treatment and nutrient removal. The resulting effluents have an excellent quality which can be improved further by applying advanced treatment processes like flocculating filtration, membrane filtration, UV or activated carbon, and others. The treated effluent can be re-used for various purposes, as process water, household water, urban water, for groundwater suppletion and drinking water. Nowadays many applications are investigated. In order to confirm the applicability pilot test investigations are done at various WWTPs. The results are promising; the cost estimations show increasing prospects. This will finally lead to the maturity of the advanced treatment. It will certainly contribute to a more sustainable water cycle. PMID- 11496664 TI - Comparison of different operational modes of a two-stage activated sludge pilot plant for the extension of the Vienna STP. AB - A pilot plant has been operated over a period of two years in order to investigate the performance and the operating characteristics of the plant concept developed for the extension of the main Vienna STP and to develop a simulation model which will be applied for operation support of the full stage plant. The pilot plant is a two stage activated sludge plant, each stage comprising of four aeration tanks and a clarifier tank. The pilot plant layout allows three different operational modes, each of which has been operated for several periods. The performance of the pilot plant during these periods is described and the different operational modes are compared to each other. PMID- 11496665 TI - Overview: full scale experience of the SHARON process for treatment of rejection water of digested sludge dewatering. AB - A SHARON system has been constructed at the Utrecht WWTP and at the Rotterdam Dokhaven WWTP. In the SHARON process rejection water from dewatering of digested sludge is treated for N-removal. It concerns a high active process operating without sludge retention. Due to differences in growth rate nitrite oxidisers can be washed out of the system while ammonia oxidisers are maintained, resulting in N-removal over nitrite. The SHARON process was selected in competition with several other techniques. The feed of a SHARON system is concentrated, with ammonia concentrations ranging from 0.5 to 1.5 g N/l. The results show that conversion rates of 90% are well possible with N-removal mainly via the nitrite route. The process was shown to be stable. Due to the high ammonium influent concentrations pH control is of great importance, preventing process inhibitions. The acidifying effect of nitrification can be compensated completely by CO2 stripping during aeration and by denitrification. Heat production by biological conversions is significant, due to the high inlet concentrations, and contributes to the optimal operating temperature of 30-40 degrees C. PMID- 11496666 TI - Nutrient removal in the river basin of the Ruhr--a German case study. AB - In the catchment area of the Ruhr, restructuring and upgrading measures in the domain of wastewater and stormwater treatment have been under way since 1990 to successively implement the currently applicable legal requirements for nutrient removal. With 2.1 million inhabitants and a design capacity of 3.7 million population equivalents (PE), it is expected that approximately DM 2 billion still have to be invested from 2000 onward. With this it will be possible to further cut the nutrient load in the Ruhr River, that has been declining consistently since the 1970s: by about 25% for Ntotal and about 10% for Ptotal. The anticipated decrease in ammonia-nitrogen in winter is particularly important for drinking water production from the river water (bank filtration). Whether and to what an extent the expected decline in phosphorus concentrations will curb eutrophication in the Ruhr with its several impounded stretches remains to be seen. Further nutrient load reductions cannot be achieved by sewage treatment related measures. Load balances underline the adverse impact of diffuse or non point sources, in particular, for nitrogen. Some potential to further improve the situation is seen in minimizing the nutrient releases from agricultural practices. PMID- 11496667 TI - The SHARON-Anammox process for treatment of ammonium rich wastewater. AB - The treatment of ammonium rich wastewater, like sludge digester effluent, can be significantly improved when new biotechnological processes are introduced. In this paper, the combination of a partial nitrification process (SHARON) and anoxic ammonium oxidation (Anammox) process for the treatment of ammonia rich influents is evaluated. Herein the combined process has been studied with sludge recycle liquor from the WWTP Rotterdam-Dokhaven. The SHARON process was operated stably for more than 2 years in a 10ICSTR under continuous aeration with a HRT of 1 day. The ammonia in the sludge liquor was converted by 53% to nitrite only. During the test period no formation of nitrate was observed. The effluent of the SHARON process was ideally suited as influent for the Anammox reactor. The Anammox process was operated as a granular sludge SBR-process. More than 80% of the ammonia was converted into dinitrogen gas at a load of 1.2 kgN/m3 per day. Planctomycete-like bacteria dominated the mixed community of the Anammox reactor, and only a small percentage of the population consisted of aerobic ammonium oxidizing bacteria. This showed that the ammonium-oxidizers in the effluent of the SHARON process did not accumulate in the SBR. The test period showed that the combined SHARON-Anammox system can work stably over long periods and the process is ready for full-scale implementation. PMID- 11496668 TI - Sludge digestion enhancement and nutrient removal from anaerobic supernatant by Mg(OH)2 application. AB - Anaerobic sludge digestion is a widely adopted process for sludge stabilization. Phosphate removal from anaerobic supernatant is necessary to limit the phosphate returned to the head of the treatment plant, thereby improving the overall treatment efficiency. In this study, magnesium hydroxide (Mg(OH)2) was used to improve the sludge digestion efficiency and to remove phosphorus from anaerobic supernatant. The anaerobic sludge digestion experiment was conducted at a pilot scale, and the results showed that applying Mg(OH)2 to anaerobic sludge digester resulted in a larger reduction in SS and COD, a higher biogas production rate, a lower level of phosphate and ammonia nitrogen concentrations in the sludge supernatant and an improved sludge dewaterability. Research results at both lab scale and pilot scale on phosphorus removal from anaerobic supernatant using Mg(OH)2 showed that a high removal of phosphorus can be achieved through the addition of Mg(OH)2. The required reaction time depends on the initial phosphorus concentration and the Mg(OH)2 dosage. PMID- 11496669 TI - Modification and expansion of a pure oxygen WWTP for biological nutrient removal (BNR). AB - A pure oxygen activated sludge system was converted to a VIP configuration BNR (biological nutrient removal) system wherein three of the five pure oxygen sections were retained, and performance was compared to that of a side-by-side air aeration MUCT (modified UCT) system. Because the pure oxygen BNR system could not obtain good nitrification, its treatment capacity had to be downgraded from 113,550 m3/d to a flow of only 60,000 m3/d. At the lesser flow, it was determined that adequate nitrification and improved denitrification could be accomplished in the pure oxygen system by continuously seeding it with 100% of the WAS from the MUCT system. Fortunately, while the capacity of the pure oxygen system had to be downgraded, it was determined that the capacity of the MUCT system was substantially greater than its design flow, and the combined system is capable of treating the entire design flow. However, this requires increasing the operating sludge age of the MUCT system. The pure oxygen BNR system performed better phosphorus removal than the MUCT system, both before and after seeding with the MUCT WAS. Apparently this was because the MUCT system was operated at a substantially higher sludge age than the pure oxygen system. However, both systems have consistently discharged effluent phosphorus concentrations of less than 2.0 mg/L TP, which is the Chesapeake Bay standard. Even with improved nitrification and denitrification in the pure oxygen BNR system, neither it nor the MUCT system have proven to be capable of meeting the Virginia Chesapeake Bay goal of 10 mg/L total nitrogen in the effluent. PMID- 11496670 TI - Surface runoff pollution by cattle slurry and inorganic fertilizer spreading: chemical oxygen demand, ortho-phosphates, and electrical conductivity levels for different buffer strip lengths. AB - As a way of dealing with the removal of pollutants from farming practices generated wastewater in the EU, we investigate the effect of spreading cattle slurry and inorganic fertiliser on 8 x 5 m2 and 8 x 3 m2 areas, referred to surface runoff chemical oxygen demand (COD), ortho-phosphates (o-P) and electrical conductivity (EC) levels, and the efficiency of grass buffer strips of various lengths in removing pollutants from runoff. The experimental plot was a 15% sloped Lolium perenne pasture. Surface runoff was generated by means of a rainfall simulator working at 47 mm h-1 rainfall intensity. Runoff was sampled by using Gerlach-type troughs situated 2, 4, 6 and 8 m downslope from the amended areas. During the first rainfall simulation, COD, o-P and EC levels were consistently higher in the slurry zone, more evidently in the larger amended area. During the second and third rainfall simulations, concentration and mass levels show a downslope drift into the buffer zones, with no clear buffer strip length attenuation. Correlation between runoff and mass drift is clearly higher in the slurry zone. Percentage attenuation in COD and o-P levels, referred to initial slurry concentrations--including rainfall dilution--were higher than 98%, and higher than 90% for EC. PMID- 11496671 TI - Nutrient removal from low strength domestic wastewater in sequencing batch biofilm reactor. AB - Nutrient removal was investigated in a packed bed column, which was operated by changing of aerated and non-aerated periods. Good removal efficiencies for nitrogen and phosphorus were obtained at long aeration period. Highest nitrification efficiency was observed in run3 because the aeration period was enough to allow nitrification. NO3 concentration was not significantly changed during the cycle, because of simultaneous denitrification during aerobic stage. Nitrogen and phosphorus removal efficiencies reached to 71 and 74% in run3 respectively. Effluent concentrations of TKN, Tot.P, NH4 and NO3 were found as 3.8, 3, 1 and 2.5 mg/l respectively. PMID- 11496672 TI - Denitrification potential enhancement by addition of anaerobic fermentation products from the organic fraction of municipal solid waste. AB - The aim of this study is to demonstrate the denitrification potential enhancement determined by the addition of the anaerobic fermentation products from the organic fraction of municipal solid waste in BNR wastewater treatment plants. The denitrification potential rose from 6 to 17 mgNO3-N/l thanks to the good performances showed by the fermentation products in the denitrification step: a typical maximum denitrification rate of about 5 mgN/gVSS d was observed. This value is similar to those obtained by using pure organic substrates. The denitrification capacity increase is determined by a significant increase of S8 and Xs availability in the influent due to the anaerobic fermentation products addition which leads to an influent COD increase equal to 68% of the total load: this value is significantly higher than the one obtained by fermentation of primary and/or waste activated sludge (3-5%). PMID- 11496673 TI - A new concept for storm water treatment: full-scale experience in Flanders. AB - Current practice in Flanders (Belgium) is to limit the hydraulic capacity of the wastewater treatment plant to 6Q14. A maximum of 3Q14 is treated in the activated sludge system, the excess flow undergoes only physical treatment (stepscreen, sand trap and settling). This paper focuses on an alternative storm management operation strategy aiming at maintaining plant performance and reducing the total pollutant discharge towards the receiving waterbody. Given the observed dilution of incoming wastewater under storm conditions, the idea was put forward that higher hydraulic loadings could be treated within the biology if additional secondary clarifier volume was supplied. The new storm operation strategy would consist of treating 6Q14 biologically using the available storm tanks as additional clarifier volume. The outcome of this study clearly shows that 6Q14 can be treated biologically using the storm tank as an extra clarifier. It was shown that doing so the overall pollutant discharge was significantly reduced. The proposed strategy does not entail any extra operational costs. On the contrary it offers a potential cost saving of 244 million Euro in view of a possible future change of environmental legislation regarding storm tank spill frequencies. PMID- 11496674 TI - Nutrient reduction policies and management strategies of the Chesapeake Bay water quality restoration program. AB - The Chesapeake Bay Program is a unique, regional, federal-state-local partnership established in 1983 by a formal Agreement between six signatories, the states of Maryland, Pennsylvania and Virginia, the District of Columbia, the US EPA, and the Chesapeake Bay Commission, to protect and restore the Bay's ecosystem. The system of governance adopted provides for dynamic interaction between the signatories, and provides for input from the interested citizens, the local governments, and the scientific and technical community. The Agreement is periodically reviewed, amended and added to by new agreements, with the most recent being the 2000 Agreement, referred to as "Chesapeake 2000: A Watershed Partnership". The new agreement, signed 28 June 2000, sets numeric goals to be accomplished during the next ten years, and includes land use policies and restrictions. PMID- 11496675 TI - Implementation of EU discharge guidelines at IVAR's Regional Wastewater Treatment Plant of North Jaeren, Stavanger, Norway. AB - Norway is a leading country on wastewater treatment comprising chemical precipitation processes. This is because Norwegian effluent standards to the North Sea have traditionally focused on phosphorus removal. In most cases, chemical treatment therefore has been considered to give lower investment and operating costs than biological treatment. Norwegian wastewater policy and management is based on the EU guidelines resulting from the EEA (European Economic Area) Agreement. According to the 1991 Urban Wastewater Treatment Directive, this will in most cases require secondary treatment. However, primary treatment can be accepted for plants larger than 10,000 PT with effluents to less sensitive coastal areas, if no negative environmental impacts can be proved. The main objective of the Regional Water, Sewerage and Waste Company (IVAR) is to comply with the prevailing effluent limits at lowest possible cost. During the past four years, IVAR has therefore undertaken comprehensive optimising of the precipitation process including full-scale experiments with different coagulant dosing control systems and different types of coagulants. IVAR also accomplished a feasibility study of introducing biological treatment as an alternative to chemical treatment. Under the prevailing frame conditions of discharge requirements and sludge deposit costs, it is not economically feasible to change to organic coagulants or biological treatment. This conclusion might have to be altered resulting from the implementation of new EU regulations and increasing sludge deposit costs. This paper presents results from full-scale experiments, extracts from the feasibility study and a comparison of costs. Furthermore, the practical consequences of implementing the EU-guidelines are discussed. PMID- 11496676 TI - Reducing phosphate discharges: the role of the 1991 EC urban wastewater treatment directive. AB - This paper reviews the status of implementation of the 1991 EC urban wastewater treatment Directive in relation to its requirements for phosphate removal from wastewater discharges. Transposition of the Directive is satisfactory in most Member States as is implementation of requirements regarding collection and secondary treatment of sewage, with the notable exceptions of Belgium and Italy. A range of approaches has been adopted for the designation of sensitive areas under the Directive and designation is still not complete. It is likely that most Member States will have met the treatment requirements for sensitive areas by the end of 1998. Exceptions will include France and Spain (where implementation will be incomplete), the UK (which designated more sensitive areas in 1998 and will meet requirements for these at a later date) and Greece and Italy (for which sensitive area designation is lacking or uncertain). The Commission has indicated that it will examine compliance for both designation and treatment closely. This may place further pressure on Member States to designate further sensitive areas. It is estimated that currently in ten EU Member States, containing 90% of the EU population, about 375,000 tonnes of phosphorus are produced in domestic wastewater each year. In 1994 39-45% of this was removed in wastewater treatment works. PMID- 11496677 TI - Modelling of full-scale wastewater treatment plants with different treatment processes using the Activated Sludge Model no. 3. AB - In 1999 the Activated Sludge Model no. 3 (ASM 3) by the IWA task Group on Mathematical Modeling for Design and Operation of Biological Wastewater Treatment was presented. The model is used for simulation of nitrogen removal. On the basis of a new calibration of the ASM 3 with the easy degradable COD measured by respiration simulation runs of this paper have been done. In 2000 a biological phosphorus removal module by the EAWAG was added to the calibrated version of ASM 3 and is now serving the current requirements for modelling the enhanced biological P-removal. Only little experiences with different load situations of large-scale wastewater treatment plants were made with both new models so far. This article reports the experiences with the simulation and calibration of the biological parameters using ASM 3 and the EAWAG BioP Module. Three different large-scale wastewater treatment plants in Germany with different treatment systems will be discussed (Koblenz: pre-denitrification; Hildesheim: simultaneous denitrification with EBPR; Duderstadt: intermediate denitrification with EBPR). Informations regarding the choice of kinetic and stoichiometric parameters will be given. PMID- 11496678 TI - The characteristics of phosphorus removal in an anaerobic/aerobic sequential batch biofilter reactor. AB - Previous research has shown that alternated anaerobic/aerobic conditions are effective in removing phosphorus from wastewater using a biofilter system. However, few studies have been conducted on the features of polyphosphate (poly P) accumulating organisms (PAOs) in biofilm on phosphorus removal. This study investigated the characteristics of the phosphorus removal mechanism in various hydraulic loads and anaerobic/aerobic time ratios using a sequential batch biofilter reactor. The storage and release of intracellular inclusions, especially polyhydroxyalkanoates (PHAs) and poly-P, would be an important factor for phosphorus removal. Under different operating conditions, total phosphorus removal was always determined by accumulation of PHAs and phosphorus release under the anaerobic phase. The PHA accumulation under the anaerobic phase was always in proportion to the biofilm phosphorus content under aerobic conditions. The result shows PAOs activity was closely related to PHA accumulation. However, the PHA accumulation under the anaerobic phase would be dependent on the hydrolysis of the complex carbon source into short chain fatty acids (SCFA). The result would be demonstrated by the simple carbon source effect. The effect of the An/Ox time ratio on TP removal was significant. Shorter anaerobic time would result in insufficient phosphorus release and greater time would result in inactive PAOs. The appropriate An/Ox time ratio was suggested as 1/2. Comparisons of the phosphorus removal characteristics between biofilm and suspended growth under the same growth conditions are discussed in detail. PMID- 11496679 TI - Contribution of P-bacteria in biological nutrient removal processes to overall effects on the environment. AB - P-bacteria can combine denitrification and P-uptake. This category of P-bacteria is abbreviated DPB. Use of DPB in BNR processes, instead of obligate aerobic PAOs, reduces oxygen consumption. Moreover, less COD is needed for the nitrogen removal. Non-required COD can be removed by presettling and used for methanation. This leads to a lower sludge production. As a result, CO2 emissions are reduced owing to less net energy consumption. Simulation for a planned WWTP with the BCFS process indicates that DPB can save 53-59% of required COD. The optimal ratios of COD/N and COD/P for simultaneous N and P removal are determined to be 3.9-4.5 and 32.2-35.2 at 12-20 degrees C. 80-95% of particulate COD can be removed from the influent, thereby CH4 production is increased by 154-271%, and the total volume of reactors can be reduced by about 50% compared to a minimised process design. Less net energy consumption over the whole WWTP contributes to a net reduction of the total CO2 emissions up to 16-21%. The energy production from CH4 is excessive enough to balance the energy consumption from aeration, dewatering and incineration. It is concluded that contribution of P-bacteria to saving COD has overall positive effects on the environment. PMID- 11496680 TI - Key tools to accelerate fulfilment of the EU Urban waste water treatment directive in the Flemish region of Belgium. AB - The EU Urban Waste Water Treatment Directive (UWWTD) has been transposed in 1992. The whole area of the Flemish region was designated as a sensitive area. This implies nutrient removal for all works in agglomerations of more than 10,000 population equivalent (PE). Thanks to an accelerated investment programme, which is in a final phase now, the wastewater treatment plants (WWTPs) will fulfil treatment in 2005. Key tools for a quick and economic execution of the programme are standardisation for new WWTP's and increasing computerisation for retrofitting existing WWTPs. The UWWTD also stipulates the reuse of treated wastewater and sludge. Strategies are explained. PMID- 11496681 TI - Acetate injection into anaerobic settled sludge for biological P-removal in an intermittently aerated reactor. AB - Injecting acetate into the sludge layer during the settling and decanting periods was adopted to enhance phosphorus release inside the sludge layer during those periods and phosphorus uptake during the subsequent aeration period in a KIST Intermittently Decanted Extended Aeration (KIDEA) process. The relationship among nitrification, denitrification and phosphorus removal was investigated in detail and analyzed with a qualitative floc model. Dependencies of nitrification on the maximum DO level during the aerobic phase and phosphorus release on residual nitrate concentration during the settling phase were significant. High degree of nitrification resulted that phosphorus release inside the sludge layer was significantly interfered with nitrate due to the limitation of available acetate and the carbon sources from influent. Such limitation was related to the primary utilization of organic substance for denitrification in the outer layer of the floc and the retarded mass transfer into the inner layer of the floc. Nevertheless, effects of acetate injection on both denitrification and phosphorus release during the settling phase were significant. Denitrification rate after acetate injection was two times as high as that before acetate injection, and phosphorus release reached about 14 mg PO4(3-)-P/g MLVSS/hr during the decanting phase after the termination of denitrification inside the sludge layer. Extremely low level of maximum DO (around 0.5 mg/L) during the aerobic phase may inhibited nitrification, considerably, and thus nearly no nitrate was present. However, the absence of nitrate increased when the phosphorus release rate was reached up to 33 mg PO4(3-)-P/g MLVSS/hr during the settling and decanting phase, and nearly all phosphorus was taken up during subsequent aerobic phase. Since the sludge layer could function as a blocking layer, phosphorus concentrations in the supernatant was not influenced by the released phosphorus inside the sludge layer during the settling and decanting period. Phosphorus removal was directly (for uptake) and indirectly (for release) dependent on the median and maximum DO concentration during the aerobic phase, and those optimal values may exist within the range from 0.2 to 0.6 mg/L and 0.4 to 1.2 mg/L, respectively. PMID- 11496682 TI - Data-based modelling and proportional-integral-plus (pip) control of nitrate in an activated sludge benchmark. AB - This paper presents the result of an investigation into the Proportional Integral Plus (PIP) control of nitrate in the second zone of an activated sludge benchmark. A data-based reduced order model is used as the control model and identified using the Simplified Refined Instrumental Variable (SRIV) identification and estimation algorithm. The PIP control design is based on the Non Minimum State Space (NMSS) form and State Variable Feedback (SVF) methodology. The PIP controller is tested against dynamic load disturbances and compared with the response of a well tuned PI controller. PMID- 11496683 TI - Development of a real-time control strategy with artificial neural network for automatic control of a continuous-flow sequencing batch reactor. AB - The purpose of this study is to develop a reliable and effective real-time control strategy by integrating artificial neural network (ANN) process models to perform automatic operation of a dynamic continuous-flow SBR system. The ANN process models, including ORP/pH simulation models and water quality ([NH4(+)-N] and [NOx(-)-N]) prediction models, can assist in real-time searching the ORP and pH control points and evaluating the operation performances of aerobic nitrification and anoxic denitrification operation phases. Since the major biological nitrogen removal mechanisms were controlled at nitrification (NH4(+)-N ->NO2(-)-N) and denitritification (NO2(-)-N-->N2) stages, as well as the phosphorus uptake and release could be completely controlled during aerobic and anoxic operation phases, the system operation performances under this ANN real time control system revealed that both the aeration time and overall hydraulic retention time could be shortened to about 1.9-2.5 and 4.8-6.2 hrs/cycle respectively. The removal efficiencies of COD, ammonia nitrogen, total nitrogen, and phosphate were 98%, 98%, 97%, and 84% respectively, which were more effective and efficient than under conventional fixed-time control approach. PMID- 11496684 TI - [Myasthenia gravis: anesthesia and the tip of the iceberg]. PMID- 11496685 TI - [Anesthesia in patients with myasthenia gravis]. AB - Myasthenia gravis is a chronic autoimmune disease characterised by progressive weakness and easy fatigability of voluntary skeletal muscles. These symptoms are related to a decrease in the number of functional acetylcholine receptors, impaired neuromuscular transmission, and a broadened neuromuscular cleft. Symptomatic treatment is based on anticholinesterases in order to increase the synaptic dwell of acetylcholine. Immune therapy includes immune suppressive drugs, plasma exchange, immunoglobulins, and thymectomy. Anticholinesterase therapy should be continued in the current mode until anaesthesia. Regional anaesthesia should be preferred. Although sensitivity to non-depolarising neuromuscular blocking agents is increased, muscle relaxants can be administered during general anaesthesia as long as neuromuscular monitoring assesses their individual effect. Due to the individual variability in the response to muscle relaxants, accurate titration in combination with pre- and intraoperative neuromuscular monitoring is essential for myasthenic patients. Postoperatively, intensive care observation is mandatory including neuromuscular monitoring. PMID- 11496686 TI - Modulation of endotoxin-stimulated TNF-alpha gene expression by ketamine and propofol in cultured human whole blood. AB - AIM: Anesthetic agents have direct and indirect effects on immunocompetent cells but the molecular mechanisms of direct interactions are largely unknown. Therefore, the effect of propofol and ketamine on TNF-alpha gene expression was studied in cultured blood from healthy volunteers. METHODS: TNF-alpha was measured in blood cultured in the absence or presence of propofol or ketamine after stimulation with E. coli endotoxin. TNF-alpha concentrations were correlated with the expression of TNF-alpha mRNA in mononuclear cells. The half life of TNF-alpha mRNA was assessed after addition of actinomycin D, an inhibitor of DNA-directed RNA synthesis. RESULTS: Propofol increased TNF-alpha mRNA (+42%) and protein (+44%) response while the addition of ketamine attenuated the TNF alpha mRNA (-31%) and protein (-53%) response. The half-life of TNF-alpha mRNA was unaffected by the anesthetics. CONCLUSION: These data suggest that propofol and ketamine have opposite effects on transcription of the TNF-alpha gene. While the effects of propofol were observed with concentrations corresponding to an induction bolus, the effects of ketamine were restricted to higher concentrations. PMID- 11496688 TI - [Ethical questions in intensive care]. PMID- 11496687 TI - [Exertion-related heat stroke. Lethal multiorgan failure from accidental hyperthermia in a 23 year old athlete]. AB - We report the case of a 23-year-old rower who suffered from an exertional heatstroke while trying to lose 2 kg in weight by jogging before a competition. The development of this illness was favoured by clothes that were inappropriate for the environmental conditions and which the sportsman wore intentionally to enhance sweating. The maximum core temperature was over 43 degrees C. As a consequence the comatose patient developed a fulminant multi-organ failure with the liver ceasing its function, renal failure, massive rhabdomyolysis, and disseminated intravascular coagulation. In addition, he suffered from pericardial effusion and acute pulmonary failure (ARDS). In spite of maximum intensive care with an extensive substitution of blood products, continuous hemodiafiltration, and inhalative administration of nitrous oxide the young sportsman died 48 h after his admission to the intensive care unit. This tragic course demonstrates the danger of the widespread habit of losing weight by vigorously exercising with inappropriate clothes. In this article, potential risk factors, symptomatology, therapy, and methods of preventing an exertional heatstroke are shown and discussed. PMID- 11496689 TI - [Biometric data on risk of pneumothorax from vertical infraclavicular brachial plexus block. A magnetic resonance imaging study]. AB - In the present study 48 sagittal and transversal magnetic resonance images of volunteers were examined for biometric data concerning risk of pneumothorax at the vertical infraclavicular blockade (VIP) of the brachial plexus. With a correct puncture the plexus can be reached after 3 cm. The shortest way to the lung is 5.3 cm (3.1-8.7 cm) at a incorrect medial angle of puncture of 46.3 degrees (35-58 degrees). While moving the angle of puncture at a minimum of 24.1 degrees (1-51 degrees) in a medial direction, a depth of 6.1 cm (4-8.9 cm) has to be reached for fatal lung puncture. The puncture point has to be determined 2.8 cm (0-4.1 cm) towards the midline of the body to have a pleura connection by a strictly vertical puncture at 6 cm (4-8.9 cm). In asthenic women, shorter distances were obtained. A considerable lower deviation can lead to pleural damage (7.5 degrees; 4.7 cm). The plexus is very close to the skin surface (1.6-3 cm). In one case, the risk for pneumothorax could be measured even with the correct puncture technique. Overall, the VIP is a very safe method for brachial plexus anaesthesia with regard to the risk of pneumothorax. In asthenic women, the risk seems to be higher but can be minimised by reducing the maximum puncture depth. PMID- 11496690 TI - [Peridural analgesia in delivery. Clarification and documentation of the obstetrical, anesthesiologic and judicial viewpoints]. PMID- 11496691 TI - [Medical standby and working hours. Judgement of the European Court of Justice of 10/3/2000 (ruling: C-303-98) on the practice of medical standby]. PMID- 11496692 TI - [100 years ago: Oskar Kreis, a pioneer in spinal obstetric analgesia at the University Women's Clinic of Basel]. AB - In 1900, Oskar Kreis (1872-1958), a gynecologist and obstetrician who received his training at the Basle University Women's Hospital, pioneered the use of spinal anaesthesia in six parturients for labour pain relief. Cocaine was used as a local anaesthetic, which had previously been shown to be effective for spinal anaesthesia by August Bier in 1898. This important advance in anaesthetic care was not widely acknowledged for a long period of time and it has only been during the past few decades that spinal anaesthesia was rediscovered as an important technique available for obstetric anaesthesia. PMID- 11496693 TI - [Diagnostic value of S-100 protein and neuron specific enolase as serum markers of cerebral disease following general anesthesia. Comment of the paper of U. Lindstedt et al. in Der Anaesthesist (2000) 49:887-892]. PMID- 11496694 TI - [Definition of normal levels of S-100. Reply to the remarks of C. Woertgen and R. D. Rothoerl]. PMID- 11496695 TI - [Endotracheal tube for pediatrics: safety examination of the anesthesia machine]. PMID- 11496696 TI - [Prevention of thrombosis]. PMID- 11496697 TI - [Heparin administration in peridural catheter anesthesia]. PMID- 11496698 TI - [Peripheral nerve block of the lower extremities]. PMID- 11496699 TI - Preventing sick buildings. PMID- 11496700 TI - Offsetting natural 'poisons'. Incontinent residents require vigilant, gentle skin care. PMID- 11496701 TI - Making a facility feel like home. PMID- 11496702 TI - Comings and goings. Transportation need not be the "T" word. PMID- 11496703 TI - Hidden costs--a real threat. Operations analysis is an imperative. PMID- 11496704 TI - Apollo CEO Raymond Nass. Interview by Suzanne Bilyeu. PMID- 11496705 TI - [Intravascular treatment with ionizing radiation for the prevention of coronary restenosis: a new story with a known personage]. PMID- 11496706 TI - [A new panorama for understanding the molecular connections between obesity and type 2 diabetes mellitus]. PMID- 11496707 TI - Persistent hypercalcitoninemia in patients with medullary thyroid cancer: a therapeutic approach based on selective venous sampling for calcitonin. AB - BACKGROUND: Persistent or recurrent medullary thyroid carcinoma (MTC) can be cured by microdissection of residual metastatic lymph nodes in the neck. Selective venous sampling can be used for localization. The aim of this study is to prospectively analyze our results with a therapeutic approach based on venous sampling, in patients with hyperthyrocalcitoninemia after thyroidectomy for MTC. METHODS: Selective venous sampling for determination of stimulated calcitonin was obtained in all patients after performing a complete laboratory and imaging workup. Patients with a gradient between the suprahepatic vein and the superior vena cava underwent unilateral or bilateral extensive lymph node dissection. We used the gradient between the right and left jugular veins to decide which side of the neck to operate. Calcitonin levels were obtained after surgery and a pentagastrin test was performed one year later if basal levels remained normal. RESULTS: Mean age of the five women with a neck gradient in the selective venous sampling who underwent neck exploration was 45 years. In all patients metastatic lymph nodes were found at the site suggested by the study. Mean positive/resected lymph nodes were 5/20. Postoperative basal and stimulated levels of calcitonin became normal in two patients at one year of follow up. CONCLUSION: Selective venous sampling is useful to localize recurrent MTC. PMID- 11496708 TI - [Descriptive analysis of 41 patients with a pyogenic hepatic abscess]. AB - OBJECTIVE: To identify the clinical, laboratory and imaging characteristics of 41 patients with a pyogenic hepatic abscess. RESEARCH DESIGN: Retrospective, descriptive. SETTING: Instituto Nacional de Ciencias Medicas y Nutricion, Salvador Zubiran. STUDY UNITS: 41 patients with a pyogenic hepatic abscess. MAIN MEASUREMENTS: The measurement of the following variables was carried out: previous background, period of evolution, symptoms, signs, laboratory studies, imaging, treatment, complications and evolution. RESULTS: 41 patients were evaluated. The average age of the group was 52.5 years (SD = 14.3) and 30 (73%) were male. The most frequent associated disease was diabetes mellitus which was found in 15 patients (37%). The most frequent clinic data were: fever in 38 patients (93%), chills in 26 (63%) and pain in the upper right quadrant in 25 (61%). The most common source of the formation of the abscess was of biliary origin in six patients (15%) and the serum amoeba test was positive in 10% of the cases. The localization of the abscess in our series was as follows: 33 cases (81%) were from the right lobe, five (12%) from the left lobe, three (7%) from both lobes and 87% were solitary. The germ found more frequently was E. coli in five patients (15%). As for the treatment, puncture by computed tomography was carried out in 25 patients (61%), four patients had a surgery and the rest were treated only with antibiotics. Regarding mortality, only one patient died (2%) due to a septic shock. CONCLUSIONS: An association with diabetes mellitus was identified and the most frequent origin was biliary, these data have already been reported in other studies. On the other hand, mortality and morbidity rates in our study were low. PMID- 11496709 TI - [Usefulness of the coccidioidin skin test in patients with type diabetes mellitus in an endemic zone]. AB - OBJECTIVE: There are few studies available about skin response to mycotic antigens in diabetes mellitus subjects, therefore, the possible difference of skin reactivity to coccidioidin in subjects with and without diabetes mellitus was analyzed. MATERIAL AND METHODS: The prevalence of skin reactivity to coccidioidin in a population sample of 1651 subjects in a coccidioidomycosis endemic zone was estimated using a transversal design. Subjects with diabetes mellitus were identified and the diagnosis was validated by clinical and laboratory criteria. In order to determine the reactivity association level with the diabetes mellitus history, data was compared with the population sample, through logistic regression analysis adjusted by age, sex and residence geographical area. Odds ratio (OR) and 95% confidence intervals were obtained. RESULTS: In the population study, there were 665 coccidioidin positive subjects (40.28% rate). Seventy six cases with diabetes mellitus were identified, 23 were positive to the test (30.26% rate) with an odds ratio of 0.63 for this group (95% CI 0.37-1.07). The OR decrease to 0.52 (95% CI 0.31-0.88, p = 0.014) with the adjusted logistic regression analysis. CONCLUSION: Coccidioidin reactivity was lower in the diabetes mellitus cases than in general population. It is necessary to be cautious with the coccidioidin test interpretation in people with DM 2. PMID- 11496710 TI - [Radiopharmacokinetic and gammagraphic studies for calculating personalized dosimetry]. AB - In nuclear medicine radiation absorbed doses are important in the patient's risk/benefit evaluation and are estimated by means of biological and complex mathematical models. The biological model includes radiopharmacokinetic data obtained through blood and urine samples taken at given intervals. A useful mathematical model is the MIRD model and with the value for the time of residence tau the MIRDOSE3 computer program uses several anatomic models and calculates radiation absorbed dose for 25 organs. At the Radiopharmacy Unit of the Nuclear Medicine Department at INCMNSZ two new bone seeking radiopharmaceuticals, 99mTc ABP and 188Re-ABP, have been designed, characterized and animal-tested. Radiopharmaceutical parameters and sequential scanning were obtained for diagnostic 99mTc-ABP in 10 normal subjects and the aim was to use % 24 hour urine elimination and % bone uptake to calculate radiation absorbed dose and extrapolate the values to 188Re-ABP as the basis for a therapeutic treatment. 99mTc-ABP was eliminated in women's urine 63.2 +/- 7.3%/activity and 70 +/- 11%/activity in men. In women 36.8 +/- 7.3% of the radiopharmaceutical remains on the bone surface and in men 30 +/- 11%. ROIs were drawn on the images and the time-integrated renal cpm/pixel/ROI gave a residence time tau = 0.52 h. Cumulative bone activity A calculated with A = 1.443 (T1/2) A0 was 2358 +/- 469 MBq h for women and 1923 +/- 707 MBq h for men. Residence time tau was 3.19 +/- 0.63 h in women and 2.6 +/- 0.95 h in men. Radiation absorbed dose for the whole body was 0.0020 +/- 0.0004 mGy/MBq for women and 0.0013 +/- 0.0005 mGy/MBq for men. For women's bone marrow it was 0.0063 +/- 0.0013 mGy/MBq and for men 0.0041 +/- 0.0015 mGy/MBq. 188Re-ABP behaves as 99mTc-ABP therefore, the effective dose given by 188Re, a beta emitter, would be for women 0.0936 mSv/MBq and for men 0.0608 mSv/MBq. These characteristics and the radionuclidic characteristics of 188Re indicate that 188Re-ABP might be a good bone metastases pain palliation radiopharmaceutical. PMID- 11496711 TI - Molecular monitoring of the treatment of patients with BCR/ABL (+) chronic myelogenous leukemia. AB - BACKGROUND: The molecular follow-up of patients with chronic myelogenous leukemia (CML) has been described as useful in other countries, but there are not data reported in Mexico. METHODS: All patients studied at Laboratories Clinicos de Puebla/Centro de Hematologia y Medicina Interna de Puebla in which the BCR-ABL hybrid gene was identified by means of polymerase chain reaction were analyzed. In 22 individuals the molecular marker of the disease was studied at diagnosis and in different instances afterwards; these patients were treated with chemotherapy, interferon, autologous or allogeneic bone marrow transplantation. RESULTS: Only the six patients that were allografted from HLA-identical siblings cleared the molecular marker of the disease; the rest of them did not achieve molecular remissions. The median survival (SV) of the whole group has not been reached, whereas the 53-month SV is 68%. One of the allografted patients died as a result of complications of graft versus-host disease. CONCLUSIONS: We have found useful the molecular monitoring of the treatment of patients with CML. Using this approach, we found that molecular remissions can be accomplished only with allografting; however, other therapeutic approaches may also result in long lasting hematologic remissions. PMID- 11496712 TI - [Behavior-immunity relationship: the role of cytokines]. AB - There are several phenomena in which the immune and the central nervous systems regulate each other. However, their mechanisms are poorly understood. Since cytokines have a central role in the regulation of the immune response, this review describes their participation in two forms of neuro-immune communication, immunomodulation by psychological stress and behavioral conditioning of immune response. The role of cytokines in the endocrine and behavioral effects of acute phase, where cytokines have an effect in functions of the central nervous system, is also reviewed. The effects of psychological stress are described as both immunosuppressing and immunoenhancing. Among them, a relevant immunosuppressing one is the reduction of IL-1, IL-2, and IFN-gamma levels. In contrast, some of the pro-inflammatory effects of stress are mediated by an increase in the levels of IL-6, IL-1, and TNF mediated by the neurotransmitter Substance P. A possible role for IL-1 and IFN-beta as possible messengers in immune regulation by behavioral conditioning is proposed. Pro-inflammatory cytokines in turn can activate the hypothalamus-pituitary-adrenal axis and induce sickness behavior during the acute phase response, during which the parasympathetic nervous system serves as pathway for their detection by the central nervous system. An account is given about recent findings on the regulation of cytokine expression by neurotransmitters from the sympathetic nervous system (epinephrine and norepinephrine), a key piece in all these mechanisms of brain-immune communication. Possible mechanisms and pathways of communication between the brain and the immune system, as well as the possible participation of other cytokines are discussed. PMID- 11496713 TI - [Familial hypercholesterolemia]. PMID- 11496714 TI - [Tumor suppressor gene p53: mechanisms of action in cell proliferation and death]. AB - Normal development is a balance process, which includes proliferation and cell death. Indeed both proliferation and apoptotic cell death are very complex process that involves the participation of many genes. In both events, the tumor suppressor p53 is one of the most important and studied genes. This transcription factor activates several genes, which results in the arrest of the cellular cycle and cellular repair or apoptosis. Many are the signals that activate p53 function including: DNA damage by gamma or ultraviolet radiation and chemical agents and hypoxia, among others. When p53 is activated it can either induces the expression of p21 (Waf1, Cip-1), which participates in the cellular arrest between G1-S transition, or the expression of bax, PIGs, IGF-BP3, Fas, FasL and DR5. The former genes participate in the cascade of events that induce apoptosis. Cellular arrest or apoptosis depends of the degree of cellular damage. The final outcome of the different mechanisms of action of p53 is to maintain the genomic stability of the cell. Thus, the absence of this protein contributes to genomic instability, the accumulation of mutations and increased tumorigenesis. It has been demonstrated that p53 present mutations in 50-55% of all types of reported human cancer. These mutations are primary located in DNA binding domain of the protein, which results in the loss of its biological activity. Frequently, tumors that present wild type p53 have a better response towards therapy than those that present p53 mutations. This review is focused on the knowledge of the normal p53 cellular pathways and their alterations in cancer. It is clear that the understanding of p53 function in the development of this pathology may give new insights in future therapeutic strategies including gene therapy for cancer. PMID- 11496715 TI - [HLA genes and the origin of Amerindians]. PMID- 11496716 TI - [The Health Boards in New Spain. 18th and 19th Centuries]. AB - OBJECTIVE: To describe the function of New Spain's Boards of Health, which were created in times of emergencies as in epidemic diseases or natural catastrophes. MATERIAL: Most of the information comes from primary sources, such as the rules, orders and reports of the Boards of Health. RESULTS: Whenever an epidemic disease appeared in New Spain, there were not enough physicians, hospitals or medical services to fulfill the needs of society. The response was the creation of Boards of Health that functioned as charitable groups. Their purpose was to provide food, clothing and medical support to the sick, by collecting donations and distributing them among the needed, and also by assigning physicians to specific geographical sectors within cities. CONCLUSIONS: The presence of Boards of Health was useful to fight epidemics in New Spain allowing the team work not only among physicians, but also among parish priests and neighborhood to offer assistance and help to sick people as medical, welfare work and soulful support. PMID- 11496717 TI - [Image of the month. Paralysis of the large hypoglossal nerve]. PMID- 11496718 TI - [How I treat ... refractory epilepsy by vagal nerve stimulation]. AB - Intermittent left vagus nerve stimulation is a novel therapeutic modality that can be proposed to patients with a refractory epilepsy and for whom resective surgery is not an option. Its precise mechanism of action is not known. Controlled studies have shown that its efficacy is similar to that of antiepileptic drugs: 50% decrease in seizure frequency in 40% patients after two years. Side effects which are generally mild to moderate are the result of a diffusion of the stimulation to the larynx. No CNS side effect has been reported. PMID- 11496719 TI - [Clinical case of the month. Biventricular stimulation, a new adjuvant therapy in the treatment of congestive heart failure]. AB - The pharmacological treatment of congestive heart failure has improved its morbidity and mortality. Nevertheless, many patients remain disabled in spite of optimal treatment. The availability of alternative therapies such as heart transplantation or cardiomyoplasty is limited. Newer devices are awaited. Biventricular pacing is a potential option to improve the homogeneity of contraction and the overall mechanical performance without myocardial oxygen consumption increase. This approach has proved acute hemodynamic and functional benefits but has to demonstrate a favourable influence on long term morbidity and mortality. PMID- 11496720 TI - [Pneumomediastinum in children]. AB - Pneumomediastinum in children is diagnosed in two circumstances: cervical subcutaneous emphysema or radiological findings. The predominant symptoms are dyspnoea, stabbing chest pain, sore throat and dysphagia. Traumatic injuries and pulmonary diseases such as asthma are the most common causes of pneumomediastinum. It may rarely result from iatrogenic manoeuvres or acidocetosis. Spontaneous mediastinal emphysema is seldom reported in children. Chest X-ray films are essential investigations. The treatment is directed towards the underlying cause, with conservative management being sufficient in most cases. However, the risk of surveying of pneumothorax or tension pneumomediastinum justifies close clinical follow-up in a specialised care unit. The onset of these pathologies necessitates a more aggressive therapy by aspiration through percutaneous catheter placed in the mediastinum. PMID- 11496721 TI - [Management of hemangiomas and vascular malformations]. AB - Vascular anomalies can be classified as hemangiomas or vascular malformations. Hemangiomas are benign neoplasms, frequently diagnosed in infancy. The vast majority of these anomalies are totally harmless and spontaneously regress, although they often worry the parents because of esthetic reasons. Only a few (1/4), however, are endangering and will require therapy. Vascular malformations are rather vessel abnormalities which, unlike hemangiomas, persist. They require a thorough evaluation and most will benefit from an intervention. The recent development of informative imaging techniques (ultrasonography, Dopplerflow imaging and magnetic resonance) has permitted significant advances in the etiology and therapy of these vascular anomalies. All too often, these patients shuffle from physician-to-physician seeking help. Thus the authors recommend a multidisciplinary approach. This team should include a plastic surgeon, a radiologist and a paediatrician. PMID- 11496722 TI - [Severity scoring systems in the the ICU. Description, utilization, and potential]. AB - Severity scoring systems are of great interest for the intensive care physicians. They provide major objective information regarding their patients and their medical practice. Scoring systems have been regularly improved over the two last decades. The different existing systems present advantages and disadvantages which influence their use in the intensive care. These scoring systems offer a wide range of analysis, however they are often underused. In this paper we suggest recommendations that could help to maximise the information provided by these systems. PMID- 11496723 TI - [Coronary revascularization of the beating heart using complete arterial grafts]. AB - A case of complete arterial revascularization using both mammary arteries and the left radial artery is reported. The operation was done on the beating heart with the adjunct of a Y graft owing to a severely calcified ascending aorta. This observation is the occasion to review the basic principles of beating heart surgery. PMID- 11496724 TI - [Biochemical mediators of inflammation]. AB - Inflammatory processes are the physiological response of the organism to different stimuli such as trauma, infections or immunological reactions. The events leading to inflammation are characterized by leukocytes adhesion to the endothelium, diapedesis and migration, cells activation and tissue remodelling. These processes are initiated and regulated by a great variety of inflammatory mediators including cytokines, prostanoids, leukotrienes, neuropeptides, reactive oxygen and nitrogen species, complement components, coagulation factors and metalloproteases. This paper is devoted to the description of the major local effects of these mediators in the inflammatory reaction. PMID- 11496725 TI - [Euthanasia and other medical decisions concerning the end of life in Belgium: epidemiologic studies]. AB - BACKGROUND: The study presented here is the first replica of the Dutch death certificate study on end of life decisions (ELDs). The main objective was to assess the incidence of euthanasia (the administration of drugs with the explicit intention to shorten the patient's life at the explicit request of the patient), physician assisted suicide (PAS), and other ELDs in medical practice in Belgium (Flanders). METHODS: A 20% random sample of 3,999 deaths was selected from all death certificates between January 1 and April 30, 1998. The physicians who signed the death certificates received one mail questionnaire per death case. FINDINGS: The response rate of the physicians was 52%. The results were corrected for the non response bias, and extrapolated to estimated annual incidences after seasonal adjustment for causes of death. It was estimated that 1.3% (1.0-1.6%, CI: 95) of all deaths resulted from euthanasia or PAS. In 3.2% (2.7-3.8%, CI: 95) of all cases, the physician ended the patient's life with lethal drugs without the explicit request of the patient. Alleviation of pain and symptoms with opioids in doses with a potential life shortening effect preceded death in 18.5% (17.3-19.7%, CI: 95) of cases and nontreatment decisions in 16.4% (15.3-17.5%, CI: 95) of cases, of which 5.8% (5.1-6.5%, CI: 95) with the explicit intention of ending the patient's life. INTERPRETATION: ELDs are prominent in medical practice in Belgium (Flanders). The incidence of deaths preceded by an ELD is similar to the Netherlands, but greater than in Australia. However, in Belgium (Flanders) the incidence of ending of patient's life without the patient's explicit request (3.2%, 2.7-3.8% CI: 95) is similar to Australia (3.5%, 2.7-4.3% CI: 95), but significantly higher than in the Netherlands (0.7%, 0.5-0.9% CI: 95). PMID- 11496726 TI - [Occupational stigmata in instrumental musicians]. AB - The frequent practice of a music instrument represents a risk for frictional or allergic contact dermatitis. The localisation and type of lesions are often typical for the musician specialty. PMID- 11496727 TI - [Pharma-clinics. Medication of the month. Repaglinide (NovoNorm)]. AB - Repaglinide (NovoNorm) is an antidiabetic oral agent of the new glinide class with insulinotropic activity. Its action on insulin secretion is more rapid and shorter than that of sulphonylurea compounds. Thanks to these properties, repaglinide is able to better control postprandial hyperglycaemia and is associated with a lower risk of delayed hypoglycaemic episodes. It is indicated for the treatment of type 2 (non-insulin-dependent) diabetes mellitus as monotherapy, after diet failure, or in combination with metformin, when the biguanide is insufficient. NovoNorm is commercialized as tablets of 0.5, 1 and 2 mg, to be taken just before each meal. Initial dosis should be 0.5 mg before meal in diabetic patients on diet alone or 1 mg before meal in patients already receiving an hypoglycaemic agent. If necessary, the dosis should be progressively increased, depending on the individual response, up to 4 mg before meal (maximal daily dosage of 16 mg), in order to optimize blood glucose control. PMID- 11496728 TI - [Pharma-clinics. Medication of the month. Montelukast (Singulair)]. AB - Montelukast (Singulair, Merck Sharp & Dohme) is a leukotriene receptor antagonist (LTRA) recommended in asthma inadequately controlled by inhaled corticosteroids, or in exercise-induced asthma. The molecule combines bronchoprotective, bronchodilating and anti-inflammatory effects. The drug has to be taken by the oral route at the dosage of 10 mg/day in adults and 5 mg/day in children under 15 years. In asthmatics montelukast decreases diurnal and nocturnal symptoms, the use of short-acting bronchodilator and improves lung function. The beneficial effects of montelukast are observed whether inhaled corticosteroids are concomitantly used or not. The drug is generally well tolerated and does not require serum monitoring. Montelukast is a complementary to inhaled corticosteroids in controlling asthma. PMID- 11496729 TI - [Clinical study of the month. Prevention of type 2 diabetes in overweight patients with impaired glucose tolerance: efficiency of lifestyle changes]. AB - The "Finnish Diabetes Prevention Study" is a prospective controlled study recently published in the New England Journal of Medicine which compared the risk of developing type 2 diabetes among 265 high-risk patients treated by changes in lifestyle to that of 257 matched controlled patients. This intervention trial demonstrated that the risk of type 2 diabetes could be reduced by 58% (p < 0.001) after a mean follow-up of 3.2 years with changes in lifestyle of high-risk overweight subjects with impaired glucose tolerance. PMID- 11496730 TI - [Cerebrogenic cardiac arrhythmia in patients with organic brain diseases of nonvascular genesis]. AB - Cardiological examination covered 115 patients (79 males and 36 females aged 16 44 years) with organic brain pathology of nonvascular genesis: epilepsy (n = 51), sequelae of closed craniocerebral trauma (n = 37) or cerebral inflammation (n = 27). The control group consisted of 15 healthy subjects. It was found that patients with brain diseases have a variety of cardiac rhythm and conduction disturbances which occur much more often than in the control group (p < 0.01). Cardiac arrhythmia was encountered most often in epileptics and patients with complications of closed craniocerebral trauma (96.1% and 70.3%, respectively). Rates of cardiac arrhythmia and conduction disorders in patients with brain diseases correlated with frequency of epileptic fits and severity of organic alterations in the brain. The pattern of rhythmic and conduction disorders was formed according to dysfunction of suprasegmentary vegetative centers with prevalent activation of the sympathetic or parasympathetic autonomic nervous system. Cardiac arrhythmia and related negative feelings deteriorate "quality of life" in patients with brain pathology that's why it is necessary to detect and treat them early. PMID- 11496731 TI - [Diagnosis of atypical clinical variants of myocardial infarction at prehospital stage]. AB - The paper presents the results of prehospital diagnosis of myocardial infarction (MI) in 1500 patients. MI was atypical in 28.6% patients. Most frequently occurred: MI with atypical pain syndrome (11.7%), asthmatic MI (7.0%), arrhythmic MI (2.9%), asymptomatic (3.3%). Late diagnosis of typical (anginal) and atypical MI was 13.4 and 21.2%. Hyperdiagnosis took place in 19.9 and 45.8% (p < 0.01), respectively. Reduction of hypo- and rise in hyperdiagnosis occurrence in 1992 1996 occurred due to atypical variants. This is explained by higher medical skills of the physicians and better knowledge about MI atypicity. PMID- 11496732 TI - [Lectin-induced platelet aggregation inpatients with unstable angina pectoris treated with anti-aggregation agents]. AB - Lectin-induced platelet aggregation was studied in patients with unstable angina pectoris treated with tiklid and aspirin. ADP, PHA-P, WGA, Con A served as the aggregation inductors. In unstable angina pectoris lectin-induced platelet aggregation was enhanced. Changes in this aggregation in response to treatment with antiaggregants with different mechanisms of action are demonstrated. PMID- 11496733 TI - [Specific features of lipid atherogenic modification in patients with ischemic heart disease and diabetes mellitus]. AB - A comparative study of the effect of concomitant compensated diabetes mellitus (DM) on plasma oxidation in patients with ischemic heart disease (IHD) with stable angina of effort (functional class II-III) in 28 anginal patients with IHD, arterial hypertension and DM, 67 anginal patients with IHD and hypertension, 57 anginal patients with IHD and 28 donors. Lipoproteins oxidation was studied by enzymic methods. A significant effect of the type of DM on dynamics of plasma oxidation in IHD patients was found. This may be an additional prognostic criterion of IHD progression in DM patients. PMID- 11496734 TI - [The role of bronchospasm in the impaired bronchial permeability in sarcoidosis]. AB - VC, FVC, FEV1, FEV1/VC%, PEF, MEF25, MEF50, MEF75 were registered in 24 sarcoidosis patients before and after inhalation of 1 dose of berotec and 1 dose of atrovent. Bronchospasm took place in 20.8% patients. Bronchospasm was the only cause of bronchial obstruction in 16.7%, one of the causes in 4.2%. Contribution of bronchospasm to development of bronchial obstruction was essential in all the examinees (91-100%). PMID- 11496735 TI - [Risk factors of progression in cryoglobulinemic glomerulonephritis associated with hepatitis C virus]. AB - The article reviews literature and original data on renal affections associated with mixed cryoglobulinemia (MCG), discuss etiology, clinical and morphological features of renal damage, variants of the clinical course and prognosis. Among 50 patients with mixed MCG selected by the presence of cryoglobulinemia (> 100 mg/dl) and in 75% with markers of hepatitis c virus infection, renal lesion was found in 14(28%). 7 of them had moderate urinary syndrome, 4 had nephrotic syndrome and 3--acute nephritic syndrome with subsequent rapid progression. 5 patients had cryoglobulinemic mesangiocapillary glomerulonephritis. A clinical case is reported illustrating complications and difficulties of treatment of cryoglobulinemic glomerulonephritis. PMID- 11496736 TI - [Relationship between duodenal ulcer recurrence rate and the degree of gastric mucosa contamination rate with Helicobacter pylori]. AB - Recurrence rate of duodenal ulcer (DU) within one and two years after treatment was studied in 42 DU patients with incomplete eradication of Helicobacter pylori (HP) and 16 DU patients with its complete eradication. It is shown that DU recurrence rate after the treatment depends on the degree of gastric mucosa contamination with HP. The risk of the DU recurrence is much higher in moderate and high contamination with HP. PMID- 11496737 TI - [Composition and role of short chain fatty acids in feces and peripheral blood serum of patients with cholelithiasis]. AB - Gas-liquid chromatography was used to estimate content of short-chain fatty acids (SCFA) in 25 patients with cholelithiasis, 32 patients with irritable colon syndrome and constipation (ICS) and 35 healthy subjects. It was found that SCFA absolute and relative concentration in the feces of cholelithiasis patients is abnormal indicating disturbance of microbiocenosis as shown by changes in functional activity of some anaerobes of the intestinal microflora participating in enterohepatic circulation of bile acids. Alterations of SCFA content in the serum of cholelithiasis patients may be related to steroids disbolism. The study of SCFA in the feces and peripheral blood serum from cholelithiasis patients is of diagnostic value. PMID- 11496738 TI - [Nutritional agents and probiotics in the treatment of intestinal insufficiency syndrome and normalization of intestinal microbiocenosis]. PMID- 11496739 TI - [Gilbert's syndrome: pathogenesis and diagnosis]. AB - Pharmacological tests, dynamic hepatoscintigraphy, spectrophotometric evaluation of serum and bile content of bilirubin fractions were performed in examination of 136 patients with Gilbert syndrome (group 1), chronic hepatitis, steatosis or hepatic cirrhosis (group 2) and 23 patients free of hepatic pathology (control group). It was established that androgenic steroids have effect on bilirubin glucuronidation. Hyperbilirubinemia in Gilbert syndrome can be caused not only inhibition of bilirubin-glucuronidation in the liver but also defective uptake of bilirubin by hepatocytes. PMID- 11496740 TI - [Efficiency of intraoral applicator UPLH-01 in snore and obstructive sleep apnea]. AB - Tolerance, subjective and objective effectiveness of domestic intraoral applicator UPLH-01 were studied in the treatment of snoring and obstructive sleep apnea (OSA). In 44 patients with uncomplicated snoring the above parameters were assessed using questionnaire, in 20 patients with uncomplicated snoring and OSA paired polysomnographic investigations were made. Tolerance of the treatment was 50%. Subjective positive effect was registered in 65% of the patients. The applicator was found effective in the majority of patients with uncomplicated snoring. In mild and moderate OSA the applicator was effective only in a few cases. In severe OSA no effect was found. Patients with chronic disorders of nasal breathing exhibited deterioration of respiration and blood saturation with oxygen. Thus nasal obstruction is a contraindication to applicator use. PMID- 11496741 TI - [C-reactive protein and defect in lipid transport blood system in patients with rheumatoid arthritis]. AB - In 88 patients with rheumatoid arthritis lipid-protein spectrum of the blood serum were determined biochemically and by means of nefelometric technique, C reactive protein (CRP) was measured with enzyme immunoassay. An increase in CRP concentration was associated with lowering of apoA1 and HDL FL, triglycerides. Thus, elevation of CRP in RA reflects not only activity of inflammation but also defects in serum lipid-transport system by the atherogenic type. PMID- 11496742 TI - [Mononitrates in therapy of ischemic heart disease polycythemia vera]. AB - Effectiveness of Mono Mac was assessed in patients with ischemic heart disease (IHD) and concomitant polycythemia vera (PV). 24-h ECG-monitoring, stress-echoCG were performed in 28 patients aged 35-79 years with painless myocardial ischemia (group 1) and painful form (group 2). Mean dose of Mono Mac was 56.4 mg/day. Clinical response was achieved. Positive changes were observed in decreased number of anginal attacks and nitroglycerin tablets, ST wave depression, number of ischemic episodes, improvement of hemodynamics. It is concluded that Mono Mac has a marked clinical effect in IHD patients having associated polycythemia vera. PMID- 11496743 TI - [Aerophytotherapy with olesan oil in the treatment of patients with chronic obstructive bronchitis and bronchial asthma]. AB - Positive clinical changes were found in 28 (64%) of 44 patients with chronic obstructive pulmonary diseases (COPD) treated with olesan-oil as aerophytotherapy. These patients improved also external respiration function. PMID- 11496744 TI - [Metadoxyl in combined treatment of alcohol damage to the liver]. AB - 20 patients of the study group and 12 control patients with alcoholic liver lesion received i.v. glucose and vitamins solution, lipotropic drugs. In addition, patients of the study group were given metadoxyl tablets (Baldacchi, Italy) in a dose 1500-2000 mg/day per os for 30 days. Before and after the treatment the patients underwent clinical examination, biochemical tests, ultrasonography, scintigraphy. It was found that metadoxyl stimulates normalization of liver function, is well tolerated, produced no side effects, holds perspectives for prevention and delay of hepatic cirrhosis. PMID- 11496745 TI - [Cough: physiological, clinical and psychological aspects]. PMID- 11496746 TI - [Progressive paralysis and antibodies to cardiolipin]. PMID- 11496747 TI - [Embolization of hepatic artery's branches for radical hemostasis in a patient with hepatic lesions]. PMID- 11496748 TI - [Ulcer and problems of Helicobacter pylori infection: new facts, reflections, suggestions]. PMID- 11496749 TI - [Use of angiotensin-converting enzyme inhibitors in chronic cardiac insufficiency (documents of XVI-XXI congresses of European Cardiologic Society)]. PMID- 11496750 TI - [Systemic manifestations of intestinal diseases]. PMID- 11496751 TI - [Pathological systems in the CNS]. PMID- 11496752 TI - [Evaluation of systemic activity of the brain by means of an artificial intelligence model]. AB - The model based on the systems representation of informational brain mechanisms reproduces the principal stages of intellectual activities: afferent synthesis, decision making, acceptor of results of actions. The imitation of behavior of subjects in the special experimental environment with the model allows quantitative estimation of parameters of systemic stages in normal subjects of different ages and subjects with different brain dysfunctions, such as arteriosclerosis, schizophrenia, etc. PMID- 11496753 TI - [Synapse architectonics of the cerebral cortex in the evolutionary aspect]. AB - The formation of synapses in ontogenesis is an important problem of neuromorphology. The paper shows that the bulk of synapses of the developing brain is formed on the basis of previous specialization of membranes. Early in ontogenesis, most formed synapses shows asymmetric contacts. In postnatal ontogenesis, the formation of synapses proceeds due to the simultaneous occurrence of specialization of membranes and synaptic vesicles, by forming a "dot" active zone. Systemogenesis, such as the general law of brain development, plays an important role in understanding the functional significance of the shown mechanisms of maturation of synapses. PMID- 11496754 TI - [Long-term correction of brain function. Prospects of immunologic approaches]. AB - Among various approaches to long-term correction of the highest functions of the brain there are two methods that are particularly promising. These included: 1) induction of autoantibodies against the enzymes involved in the metabolism of neuroregulators by means of immunization of respective heterologous enzymes; 2) immunization by covalent conjugates of monomolecular neurotropic compounds and neuropeptides with antigen carriers. The investigations of both methods are reviewed and illustrated in experiments on albino rats during alcoholization and some other processes of pathological behavior formation. Evidence is provided for that behavior can be corrected for several months or longer. PMID- 11496755 TI - [Dynamic stereotype as a systemic property of the brain]. AB - Based on the theory of functional systems, the author has developed I. P. Pavlov's idea on the dynamic stereotype--"reality imprints". The parameters of reality, and the results of actions in particular, that meet the leading requirements of living organisms are shown to be dynamically imprinted as molecular engrams of an acceptor of results of actions on the structures involved in dominating motivation. Dominating motivations play a key role in extracting early formed acceptor engrams of results of activities in different functional systems. PMID- 11496756 TI - [Molecular genetics of brain development and learning: on the way to synthesis]. PMID- 11496757 TI - [Protein-peptide complexes in the mechanisms of inborn and learned behavior patterns]. AB - Protein-peptide complexes involved in learned and natural drinking behavioral patterns were comparatively analyzed in rats. Active immunization with protein conjugated angiotensin II and beta-endorphin produces some behavioral responses. It is suggested that protein complexes of these peptides play a specific informational role in the systemic organization of learned behavior. The paper discusses whether these complexes perform an important function due to their conformational properties and their participation in the hierarchical organization of integrative processes in the nervous system. PMID- 11496759 TI - [Mechanisms for encoding positional information in embryonic shaping of the vertebrate brain]. AB - The mechanisms of early embryonal shaping of the brain in man and animals were studied. Analysis of the biomechanical properties of development of nervous tissue and embryological experiments demonstrated that tangential neuroepithelial intention is the major source of positional information. Experimental changes in the neuroepithelial intention system resulted in various types of embryonal anomalies of the nervous system. Mechanism-dependent ion channels that have marked periods of sensitivity and determine the histogenetic direction of neuroblast cell differentiation were found to underlie the mechanosensitivity of the neuroepithelium. Experimental findings were compared with unique autopsy data on early development of the human brain. Human embryos were examined from neurulation to week 6 of development. Different types of human embryonal brain anomalies were shown to occur with 3 types of neurulation disorders: 1) an open preneuropore is responsible for anomalies of the forebrain and ethmoidal area; 2) arrested neurulation in the postneuropore leads to anomalies of the diencephalon, midbrain, and occipital region; 3) impaired neurulation in the caudal region is a cause of spinal cord anomalies. The above anomalies resulted from local compensatory responses of the neuroepithelium due to the lack of intentions that are characteristic of normal development of the neural tube. PMID- 11496758 TI - [Protein cytochemistry in revealing the essentials of the structural and functional organization of the brain]. AB - The histochemical method developed by the author was used to localize lysine- and arginine-rich proteins in the rat, rabbit, feline, canine, and human brains. A great difference was found between neurons (enlarged in lysine-rich proteins) and neuroglia (abundant in arginine-rich proteins). It was suggested that arginine rich neuroglia might be responsible for or involved in supplying the brain with L arginine, a precursor of nitric oxide (NO), a secondary messenger that is likely to be used by ancient type neurons. A direct relationship was established between the complexity of the brain structural and functional organization and the increases in lysine-rich neuronal proteins. This assumes that, by analogy with L arginine synthesis of NO, L-lysine can be a precursor of a hypothetical messenger used by neurons of the higher brain regions. Responses of neurons to experimental exposures were assessed by the changes occurring in the levels of protein and its synthesis. Responses of neurons in the cortex and subcortical areas of the brain were shown to be non-specific and characterized by automatism, but to have specific features in different types of neurons. PMID- 11496760 TI - [Prerequisites for structural asymmetry of Broca's motor speech area of the human brain]. AB - Structural features of the Broca's speech-motor area in the left and right human cerebral hemispheres were studied. There were some peculiarities in the localization, shape, and structure of the main sulci and gyri of the frontal region in different hemispheres of the human brain. There were differences in the volume of cortical areas 44 and 45 in the left and right hemispheres, as well as in the count of neurons and in the size of pyramidal neurons in cytoarchitectonical layers III and V in the left and right hemispheres of speech motor areas. It is suggested that left hemispherical asymmetry of Broca's area correlates with the development of human speech functions and oratory. PMID- 11496761 TI - [Viral diseases in humans]. PMID- 11496762 TI - [Diagnostic techniques in viral diseases]. PMID- 11496763 TI - [Current aspects of chemotherapy of sexually transmitted viral diseases]. AB - In recent years, among all sexually transmitted infectious agents, viral infections were indisputably at the head. In this group, genital herpes is one of the substantial infections. Acyclovir, valcyclovir and famcyclovir are the compounds applied in the treatment of Herpes simplex infections. In the therapy with an application of alternative drugs, the possibility of foscarnet should be inquired. These drugs have precise documentation on an application in the treatment of herpetic symptoms of acute infections. However, they do not influence, in radical way, the prevention of relapse of the disease. In spite of their universal and generally accessible application, they also do not influence the interruption of transmission of infection. It seems, that in the nearest perspective, there are hopes associated with introduction of effective and active immunization or, less probable, an introduction of the drug or method of the treatment terminating latency. PMID- 11496764 TI - [Immunoprophylaxis: prognosis and reality]. PMID- 11496765 TI - [Sexually transmitted and congenital viral diseases caused by herpesviruses]. PMID- 11496766 TI - [Epidemiology of HIV-related sexually transmitted diseases]. PMID- 11496767 TI - [The role of gastric inflammatory viruses in sexually transmitted diseases]. PMID- 11496768 TI - [New possibilities of immunization in the prophylaxis of hepatitis b]. PMID- 11496769 TI - [Etiological pathogenesis of cervical cancer]. PMID- 11496770 TI - [Clinical analysis of sexually transmitted viral diseases]. PMID- 11496771 TI - [Animal diseases spread by food]. PMID- 11496772 TI - [In vitro diagnosis of human rabies in Poland]. PMID- 11496773 TI - [Genetic and antigen relationship between protective strains and street strains]. PMID- 11496774 TI - [Raccoon (Procyon lotor) rabies outbreak in New York State]. PMID- 11496775 TI - [Pathogenesis of hemorrhagic fever]. PMID- 11496776 TI - [Hantavirus-related hemorrhagic fever in Europe]. PMID- 11496777 TI - [The role and goals of The Regional Center "Salmonella"]. PMID- 11496778 TI - [Clinical picture of certain hemorrhagic fever diseases]. PMID- 11496779 TI - [Epidemiology of Lyme borreliosis in Poland]. PMID- 11496780 TI - [Mosquitoes as vectors of arboviruses in Poland]. PMID- 11496781 TI - [Animal borreliosis]. PMID- 11496782 TI - [The kind of pathogens which could be used in the case of biological terrorism]. PMID- 11496783 TI - [Current threat: anthrax]. PMID- 11496785 TI - [Rickettsiae: potential biological terrorism]. PMID- 11496784 TI - [Sausage poison as a medium for biological terrorism]. PMID- 11496786 TI - [Escherichia coli O157 among the cattle on the Polish territory]. PMID- 11496787 TI - [The possibilities to identify biological threats]. PMID- 11496788 TI - [Discovering the results of biological terrorist attack by pyrolysis gas chromatography]. PMID- 11496789 TI - [How to handle a biological terrorist attack]. PMID- 11496790 TI - [Special military operations in preparation to the liquidation of a massive biological terrorist attack]. PMID- 11496791 TI - [Health threat of acquiring animal diseases transmitted by animals or materials of animal origin]. PMID- 11496792 TI - [Legal, organizational and diagnostic basis of animal control]. PMID- 11496793 TI - [Fundamentals of the control over the import of animals and animal products]. PMID- 11496794 TI - [Animal parasitic diseases spreading through animals in Poland: the diagnostic methods in parasite diseases]. PMID- 11496795 TI - [The current situation of investigation of bovine spongiform encephalopathy]. PMID- 11496796 TI - [Epidemiological situation of pathogens of food-related diseases (salmonellosis)]. PMID- 11496798 TI - [Spongiform encephalopathy ( prion diseases) in people]. PMID- 11496797 TI - [Prions: a new contamination factor]. PMID- 11496799 TI - [Spongiform encephalopathy (prion diseases) in animals]. PMID- 11496800 TI - [Clinical diagnosis of Creutzfeldt-Jakob disease and related diseases]. PMID- 11496801 TI - [Epidemiological forecasting of prion diseases in Poland]. PMID- 11496802 TI - [The role of meat-bone flour in the transmission of bovine spongiform encephalopathy (BSE)]. PMID- 11496803 TI - [Rabies vaccination in Poland in years 1990-1999]. PMID- 11496804 TI - [The contracting of rabies among people in Poland]. PMID- 11496805 TI - [Fading optimism when it comes to xenotransplantation. Still a long way to go before whole porcine organs can be transferred to human beings]. PMID- 11496806 TI - [Do measure bone density more often!]. PMID- 11496807 TI - [A good combination to achieve a reduced number of fractures: bone density measurements, drug therapy and hip protector]. PMID- 11496808 TI - [Count on your beliefs. Bayes--not the P value--measures credence]. AB - This article (the second of two) describes traditional (frequentistic) statistical analysis in the context of the confidence interval. Bayesian analysis is described in two settings. In the choice between diagnostic alternatives, the bayesian approach offers useful integration of new information with previous knowledge. With regard to the evaluation of clinical trial data, this article exemplifies bayesian analysis as contrasted with traditional analysis, and advantages of the former are cited. A brief literature review exposes the wide applicability of bayesian analysis in medical statistics. PMID- 11496809 TI - [Alarm defends its position. Comments to meta-analysis of alarm treatment of nocturnal enuresis]. AB - Glazener and Evans have, on behalf of the Cochrane Library, performed a systematic literature review of alarm interventions in nocturnal enuresis. An extensive, and probably exhaustive, search yielded 22 studies which were of sufficient methodological quality and in which the enuresis alarm was included as one treatment alternative. From these studies it could be concluded that 1) the enuresis alarm is clearly more effective than non-treatment, 2) different types of enuresis alarms do not differ significantly as regards efficacy, and 3) alarm treatment is not clearly superior or inferior to pharmacological treatment. PMID- 11496810 TI - [Early potty training advantageous in bladder dysfunction. Decreases the risk of urinary infection]. AB - The time when we start potty training has become increasingly postponed. This paper discusses the possible negative consequences of this social situation. Depending on the child's maturity, bladder control could be defined as anything from "continence with support of an adult before the age of one year" to "independent social control, unobtainable in our culture before the child is 4 years of age". In toddlers suffering from bladder dysfunction, the impact of potty training improves bladder emptying, decreasing residual urine and resultant urinary tract infection. In toddlers with anatomical and functional anomalies of the urinary tract, early potty training is recommended. PMID- 11496811 TI - [Osteoporosis among the elderly is a "silent epidemics". Insufficient prophylaxis and underdiagnosing are big problems]. AB - The increasing number of fractures due to osteoporosis continues to put economical and practical burdens on the national health service. Despite the fact that knowledge of the disease and its consequences has increased tremendously and new, effective drugs have been developed only a minority of the patients are diagnosed and appropriately treated. Resources should be made available to improve methods of measuring bone mineral density as well as establishing outpatient osteoporosis clinics at all hospitals in the country. Age ought not to be a limiting factor to initiate treatment or prevention of fractures. The present situation for the osteoporosis patient, the clinical syndromes with special reference to age-related and secondary osteoporosis, their causes, diagnosis and treatment are briefly reviewed. PMID- 11496812 TI - [Estrogen therapy in climacteric. Current views in an evidence-based perspective]. AB - In Sweden, approximately 65,000 women enter menopause each year. Hormone replacement therapy (HRT) for the treatment of climacteric symptoms is well documented, but the role of HRT in the prevention of other diseases is more questionable. The existing literature is discussed from an evidence-based perspective. There is strong evidence supporting the treatment of subjective symptoms, and also a scientific basis for long term treatment of women at increased risk of osteoporosis. Few studies address the issue as to how HRT should be prescribed and followed-up. It is important to consider the issue of quality of life in times of limited resources. PMID- 11496813 TI - [Transplantation of stem cells from umbilical cord blood. Important to report all results to international registries]. PMID- 11496814 TI - [The best way to learn consultation skills is through tutoring in clinical situations. Experiences from a course in Gothenburg]. AB - In 1993, a ten-week course called 'Consultation knowledge' started in undergraduate medical education in Goteborg. At the beginning of clinical clerkships students learn communication skills, clinical examination skills and documentation in a clinical context. Tutors were educated and supervised and also participated in the practical examination. Feedback from students was obtained from written evaluations and analysed. Reports from teachers' follow-up meetings were also used. Feedback data functioned as an instrument in evaluation and for development of the course. Learning objectives and core content were made clear by refining the examination and by structured support to tutors. The advantages of repeated consultation skills training in the clinical curriculum are discussed. PMID- 11496816 TI - [The methadone program in Stockholm is repressive]. PMID- 11496817 TI - [Evidence-based medicine: ten critical objections against a valuable method]. PMID- 11496818 TI - [Sweden should respect other countries' approach to the question of abortion]. PMID- 11496819 TI - [Sell paracetamol in smaller packages!]. PMID- 11496820 TI - [Final reply 1: Circumcision, citations from the Bible and scientific credibility]. PMID- 11496821 TI - [Final reply 2: Freedom of religion must not hurt the children]. PMID- 11496822 TI - [Histamine produced by macrophage and T lymphocyte: a new type of signal transducer]. AB - Macrophages (M phi) produce histamine (Hm) when activated by bacterial endotoxin (LPS) through induced histidine decarboxylase (HDC). Among the cytokines tested, GM-CSF or IL-3 specifically augmented the LPS-dependent HDC induction by M phi. Hm formed by M phi regulates synthesis of cytokines such as IL-1, IL-6, G-CSF and M-CSF by the cells per se and may modulate immune reactions and division and differentiation of various hematopoietic cells. Kupffer cells, M phi-like cells in the liver, also synthesize Hm in mice injected with hepatotoxins such as tetradecanoylphorbol acetate or LPS. Hm thus produced by Kupffer cells may participate in the regeneration of the injured liver through induction of hepatocyte growth factor. Concanavalin A (Con A) enhanced Hm formation by T lymphocytes. GM-CSF or IL-3 also enhanced the Hm synthesis by CD4+ and CD8+ T cells. Hm formed by T cells regulates immune reactions such as lymphocyte blastogenesis. In animals infected with gram(-) bacteria Hm is produced by the M phi-T cell system and may regulate immune competence to the bacteria. In addition, Hm may act as a signal transducer between the peripheral immune system and hypothalamus-pituitary-adrenal system, leading to GC secretion, in order to prevent occurrence of tissue injury caused by excess immune reactions. PMID- 11496823 TI - [Regulation of histamine production in macrophages]. AB - Stimulating cells of the mouse macrophage-like cell line RAW 264.7 with the Ca(2+)-ATPase inhibitor thapsigargin increased histamine production. Thapsigargin increased the levels of histidine decarboxylase (HDC) mRNA at 4 h and the expression of 74-kDa HDC protein at 8 h. PD98059, a specific inhibitor of MEK-1 which phosphorylates p44/p42 MAP kinase, strongly suppressed the thapsigargin induced histamine production, the increase in HDC mRNA level and 74-kDa HDC protein expression. In contrast, SB203580, an inhibitor of p38 MAP kinase, showed only a partial inhibition of histamine production. TPA and LPS also induced histamine production in RAW 264.7 cells, and the histamine production induced by TPA or LPS was also inhibited by PD98059, but the effect of SB203580 was partial. The synthetic glucocorticoid dexamethasone inhibited thapsigargin-induced histamine production, 74-kDa HDC protein expression and the activation of p44/p42 MAP kinases. In conclusion, the increase in histamine production in macrophages stimulated with inflammatory stimulants is due to the increased expression of 74 kDa HDC, which is positively regulated by activated p44/p42 MAP kinases. Dexamethasone inhibits thapsigargin-induced HDC protein expression and histamine production by inhibiting the MAP kinase activation. PMID- 11496824 TI - [Regulation of cytokine production by histamine through H2-receptor stimulation]. AB - Histamine is a well known mediator of inflammation including the allergic reaction. Histamine has been suggested to be a immunomodulator. Recent studies revealed that induction of histidine decarboxylase occurs by the stimulation of several cytokines and LPS, suggesting an immunomodulatory role of the inducible histamine. Using human PBMC culture, it was demonstrated that histamine was a potent inducer of IL-18, IFN-gamma in human PBMC. Histamine did not induce the production of IL-12. The effects of histamine on cytokine production were mimicked by H2-selective agonists and inhibited by H2- but not by H1- and H3 antagonists, indicating the involvement of H2-receptors in histamine action. All effects of histamine were abolished by the presence of anti-IL-18 antibody or IL 1b-converting enzyme/caspase-1 inhibitor, indicating that histamine action is dependent on mature IL-18 secretion and that IL-18 production was present most upstream of the cytokine cascade triggered by histamine. Histamine is a very important modulator of Th1 cytokine production in PBMC and is quite unique in triggering the cytokine cascade without inducing IL-12 production. PMID- 11496825 TI - [Identification and characterization of histamine H4 receptor]. AB - Recently, we and other groups have identified cDNA encoding the novel histamine H4 receptor. All of the groups have initially found a clue for the H4 receptor nucleotides sequence in the human draft genomic DNA database. The primary structure of H4 receptor reveals the highest homology with H3 receptor among known G-protein coupled receptors (37.4%). H4 receptor binds to histamine with high affinity, which results in the down-regulation of intracellular cAMP level. H4 receptor is activated not only by histamine, but also R-(alpha) methylhistamine (H3 receptor agonist), clobenpropit (H3 receptor antagonist), clozapine (neuroleptic) and other histaminergic compounds, while it is antagonized by thioperamide (H3 receptor antagonist). The H4 receptor is localized in the peripheral blood leukocytes, spleen, thymus, small intestine, colon, bone marrow and so on. The tissue distribution of the H4 receptor and known physiological function of histamine tempts us to speculate about its function as an immune modulator. Although there needs much additional work on characterization of the H4 receptor, the discovery of this receptor subtype will unveil a new phase for determining the physiological role of histamine. PMID- 11496826 TI - [Primary sensory neurons expressing histamine H1-receptor mRNA]. AB - Pharmacological studies have suggested that a subgroup of primary sensory neurons is responsive to histamine via the H1 receptor. However, which type of primary sensory neurons express H1 receptor is not known. We addressed this issue using in situ hybridization histochemistry with a cRNA probe for the guinea pig H1 receptor mRNA. H1 receptor mRNA was expressed in about 15-20% of the trigeminal and lumbar dorsal root ganglion (DRG) neurons, but none of the nodose ganglion neurons. The positive neurons in DRG were exclusively small in size and were labeled by isolectin B4, suggesting that these neurons have unmyelinated fibers. However, H1-receptor mRNA-expressing DRG neurons were not immunoreactive to substance P (SP) or calcitonin gene-related peptide (CGRP), which are implicated in the nociceptive transmission of the primary sensory system. Moreover, in guinea pigs neonatally treated with capsaicin (50 mg/kg), few CGRP-immunoreactive neurons were seen in DRG, but the percentage of H1-receptor mRNA-expressing neurons (15%-20%) and the intensity of the mRNA signals in these neurons were not affected by neonatal capsaicin treatment, suggesting that H1 receptor-expressing neurons are not sensitive to capsaicin. These findings suggest that H1-receptor expressing neurons are involved in the transmission of a unique sensory modality such as itch. A marked increase in the number of mRNA-positive DRG neurons was observed 1-5 days after a crush injury of the sciatic nerve (3-4-fold of the control value). These neurons that turned mRNA-positive after the nerve crush were also mainly small-sized. The mRNA signals were detected in many peptidergic (SP/CGRP) neurons, in contrast to the normal condition. On the other hand, mRNA signals were decreased in the neurons that showed intense labeling in the normal condition. These results suggest that the gene expression of H1 receptors up regulated in injured afferents may be involved in neuropathic pain. PMID- 11496827 TI - [Induction of histidine decarboxylase in inflammation and immune responses]. AB - Histamine is a classical, but still interesting inflammatory mediator. Many people have long believed that histamine is derived from mast cells or basophils alone. However, the histamine-forming enzyme, histidine decarboxylase (HDC), is induced in a variety of tissues in response (i) to gram-positive and gram negative bacterial components (lipopolysaccharides, peptidoglycan, and enterotoxin A) and (ii) to various cytokines (IL-1, IL-3, IL-12, IL-18, TNF, G CSF, and GM-CSF). HDC is induced even in mast-cell-deficient mice. The histamine newly formed via the induction of HDC is released immediately and may be involved in a variety of immune responses. Reviewing our work and that of Schayer and Kahlson, the pioneers in this field, lead us to the conclusion that nowadays we need to understand that histamine can be produced via the induction of HDC by a mechanism coupled with the cytokine network. We call this histamine "neohistamine", to distinguish it from the classical histamine derived from mast cells or basophils. Neohistamine is involved in physiological reactions, inflammation, immune responses and a variety of diseases such as periodontitis, muscle fatigue (or temporomandibular disorders), stress- or drug-induced gastric ulcers, rheumatoid arthritis, complications in diabetes, hepatitis, allograft rejection, allergic reactions, tumor growth, and inflammatory side effects of aminobisphosphonates. PMID- 11496828 TI - [Pharmacological, pharmacokinetic and clinical properties of olopatadine hydrochloride' (olopatadine), an antiallergic drug]. AB - Olopatadine is a selective histamine H1-receptor antagonist possessing inhibitory effects on the release of inflammatory lipid mediators such as leukotriene and thromboxane from human polymorphonuclear leukocytes and eosinophils. Olopatadine also inhibited the tachykininergic contraction in the guinea pig bronchi by prejunctional inhibition of peripheral sensory nerves. Oral administration of olopatadine inhibited passive cutaneous anaphylaxis in rats, experimental allergic rhinitis and bronchial asthmatic responses in actively sensitized guinea pigs. Olopatadine exerted no significant effects on action potential duration in isolated guinea pig myocardium and ventricular myocytes. Olopatadine was highly and rapidly absorbed in healthy volunteers. The urinary excretion of olopatadine accounted for not less than 58% and the contribution of metabolism was low in the elimination of olopatadine. Olopatadine was shown to be useful for the treatment of allergic rhinitis and chronic urticaria in double-blind clinical trials. Olopatadine was approved in Japan for the treatment of allergic rhinitis, chronic urticaria, eczema dermatitis, prurigo, pruritus cutaneous, psoriasis vulgaris and erythema exsudativum multiforme in December, 2000. PMID- 11496829 TI - [Glimepiride (Amaryl): a review of its pharmacological and clinical profile]. AB - In Type 2 diabetes, it is considered that the lowered insulin secretion and the lowered insulin sensitivity cause hyperglycemia. Sulfonylureas have strong blood glucose lowering effect by stimulating insulin secretion and have been widely used in the treatment of Type 2 diabetes. However, the use of sulfonylurea has several problematic issues (weight gain, hypoglycemia, second failure and so on), which would due to stimulation of strong insulin secretion. Glimepiride, a new sulfonylurea, has a blood-glucose lowering effect as strong as those of existing sulfonylureas, but only induces mild insulin secretion. The sulfonylurea receptor has a weaker affinity for glimepiride than glibenclamide. The association and dissociation to the sulfonylurea receptor of glimepiride are faster than those of glibenclamide. Additionally, it is confirmed by basic studies that part of the glimepiride effect is attributable to improving insulin sensitivity. Glimepiride has already been used in more than 60 countries in the world. Outside of Japan, several clinical studies have demonstrated that glimepiride shows less hypoglycemia and no weight gain. Glimepiride is expected to be a new efficient agent for the treatment of Type 2 diabetes. PMID- 11496830 TI - Reduced protein adsorption and platelet adhesion by controlled variation of oligomaltose surfactant polymer coatings. AB - A series of oligomaltose surfactant polymers were prepared by the simultaneous coupling of hydrophilic maltolactone [of 2(M2), 7(M7), or 15(M15) glucose units] and hydrophobic N-(hexanoyloxy)succinimide (Hex) groups to the amino groups of a poly(vinyl amine) backbone. The surfactants were characterized by FTIR and 1H-NMR spectroscopies for purity and composition. Contact-angle and AFM measurements confirmed full monolayer adsorption for all surfactants on a model surface, highly oriented pyrolitic graphite, while full coverage was observed on polyethylene only for PVAm (M7:Hex) due to the optimal M7:Hex ratio and Hex chain density. On graphite, protein resistance increased with increasing coating thickness from 81.4 to 85.8 to 95.8% for the M2, M7, and M15 surfactants, respectively. Additionally, static platelet adhesion on all three surfactants dropped substantially to 15% (M2), 17% (M7), and 16% (M15)compared to glass (adhesion normalized to 100%) and a polyurethane (24%) surface. Protein- and platelet-resistant properties of the controlled oligomaltose layers are discussed by analysis of molecular modeling, oligomaltose and hexanoyl chain densities, and surfactant stability. PMID- 11496831 TI - Protease inhibitors, part 13: Specific, weakly basic thrombin inhibitors incorporating sulfonyl dicyandiamide moieties in their structure. AB - A series of compounds has been prepared by reaction of dicyandiamide with alkyl/arylsulfonyl halides as well as arylsulfonylisocyanates to locate a lead for obtaining weakly basic thrombin inhibitors with sulfonyldicyandiamide moieties as the S1 anchoring group. The detected lead was sulfanilyl dicyandiamide (K1 of 3 microM against thrombin, and 15 microM against trypsin), which has been further derivatized at the 4-amino group by incorporating arylsulfonylureido as well as amino acyl/dipeptidyl groups protected at the amino terminal moiety with benzyloxycarbonyl or tosylureido moieties. The best compound obtained (ts-D-Phe-Pro-sulfanilyl-dicyandiamide) showed inhibition constants of 9 nM against thrombin and 1400 nM against trypsin. pKa measurements showed that the new derivatives reported here do indeed possess a reduced basicity, with the pKa of the modified guanidine moieties in the range 7.9-8.3 pKa units. Molecular mechanics calculations showed that the preferred tautomeric form of these compounds is of the type ArSO2N=C(NH2) NH-CN, probably allowing for the formation of favorable interaction between this new anchoring group and the active site amino acid residue Asp 189, critical for substrate/inhibitor binding to this type of serine protease. Thus, the main finding of the present paper is that the sulfonyldicyandiamide group may constitute an interesting alternative for obtaining weakly basic, potent thrombin inhibitors, which bind with less affinity to trypsin. PMID- 11496832 TI - Potential of pyrazolooxadiazinone derivatives as serine protease inhibitors. AB - As a part of an investigation on molecular hybrids as new serine protease inhibitors, the pyrazolo [4,3-c][1,2,5]oxadiazin-3(5H)-one ring system was selected as a model of potential mechanism-based inhibitors. Due to the inherent reactivity of this system an optimal balance between susceptibility to nucleophilic attack and stability in solvents was sought prior to development as therapeutic agents. Substitutions on N5 and C7 of the supporting pyrazole ring with either aliphatic or aromatic groups (compounds 2 a-m) and the replacement of the carbonyl oxygen on the reactive oxadiazinone ring with sulfur (compounds 3a,i) were explored. Two members (2i and 2k) of this class of inhibitors displayed time-dependent inhibition of HLE suggesting mechanism-based inhibition. The observation that HLE generated a product(s) from compound 2i which displayed an identical UV-Visible spectrum to that observed during non-enzymatic hydrolysis further supports this proposal. FlexX-based docking of these compounds into a model of the human leukocyte elastase (HLE) active site produced a molecular model of the inhibitor-enzyme interaction. PMID- 11496833 TI - Inhibitors of human and rat testes microsomal 17beta-hydroxysteroid dehydrogenase (17beta-HSD) as potential agents for prostatic cancer. AB - In a screening programme for inhibitors of human testis 17beta-hydroxysteroid dehydrogenase (17beta-HSD type 3), as potential agents for the treatment of hormone-dependent prostatic cancer, we have used crude human testis microsomal 17beta-hydroxysteroid dehydrogenase as a convenient source of the enzyme. Crude human enzyme was shown to have a similar substrate profile to recombinant Type 3 17beta-HSD from the same source as determined by the low Km/Vmax ratio for the reduction of androstenedione compared to the oxidation of testosterone, and a low level of activity in reduction of oestrone. Screening of a wide range of compounds of different structural types as potential inhibitors of the microsomal enzyme in the reduction step revealed that certain p-benzoquinones and flavones/isoflavones were potent inhibitors of the enzyme, diphenyl-p benzoquinone (2.7 microM), phenyl-p-benzoquinone (5.7 microM), 7-hydroxyflavone (9.0 microM), baicalein (9.3 microM) and biochanin A (10.8 microM). Some structure-activity relationships within the flavone/isoflavone series are discussed. Studies with rat testis microsomal 17beta-HSD showed that it differed from the human enzyme mainly in its greater ability to accept oestrone as substrate and the pH-optimum for oxidation of testosterone. It was found to be much less sensitive to inhibition by the compounds studied so negating it use as a more readily available tissue for the screening of potential inhibitors. PMID- 11496834 TI - Stable expression of the human 5alpha-reductase isoenzymes type I and type II in HEK293 cells to identify dual and selective inhibitors. AB - A eucaryotic cell assay was established to identify novel, dual and selective inhibitors of human 5alpha-reductase. For this purpose the cDNAs encoding 5alpha reductase type I and type II were inserted into a pRcCMV vector and expressed in human embryonic kidney (HEK293) cells. Single cell clones with substantially high enzymatic activity were selected and established as permanent cell lines. KM values were determined for both isozymes. The inhibitory potency of several steroidal and non-steroidal compounds synthesized in our group, as well as finasteride and 4MA as controls, were tested by measuring the conversion of [3H]androstenedione. Reaction products were quantified by a HPLC reversed phase technique. Using the new cell assays, selective as well as novel dual 5alpha reductase inhibitors with IC50 values between 1.0 and 2.5 microM were identified. PMID- 11496835 TI - Arylsulfonyl-N,N-dialkyl-dithiocarbamates as tumor cell growth inhibitors: novel agents targeting beta-tubulin? AB - Reaction of sodium N,N-dimethyldithiocarbamate or N,N-diethyldithiocarbamate with arylsulfonyl halides afforded a series of arylsulfonyl-N,N-dialkyl dithiocarbamates. The reactivity of these new derivatives with cysteine and glutathione has been investigated in order to identify derivatives that might label a cysteine residue of the heterodimeric protein tubulin which plays a critical physiological function in cell division and also possesses enzymatic activity as a GTP-ase. Since many antitumor drugs exert their action by binding to tubulin, inhibiting in this way microtubule association and provoking cell death, some of the most reactive compounds against the thiol reagents found in this work have been assayed for their antitumor activity. Indeed strong tumor cell growth inhibitory properties against several leukemia, non-small cell lung, ovarian, melanoma, colon, CNS, renal, prostate and breast cancer has been found in vitro for some of the 4-halogeno-, 4-methyl- or 4-carboxyphenyl-substituted arylsulfonyl-N,N-dialkyl-dithiocarbamates. Furthermore, some of these derivative were shown to act as in vitro tubulin polymerization inhibitors using a turbidimetric assay. PMID- 11496836 TI - Effect of cadmium acetate on the conformation of lysozyme: functional implications. AB - Structural variations of lysozyme as a consequence of its interaction with CdAc2, as well as the implications on the protein functionality have been studied. Variations in the conformation of the macromolecule are seen, however these changes are not reflected on the secondary structure. The interaction of the salt with the polypeptide chain is weak and thermodynamically unfavourable. Molecular aggregates (dimer forms) are observed at the highest salt concentrations. This interaction causes an inhibitory effect on lysozyme, the activity loss being 50% at the highest salt concentration studied. The inhibition is of mixed type with an uncompetitive component. Thus cadmium does not bind to the active site of the enzyme which is in accordance with the not very large activity loss observed. The substrate inhibition of lysozyme is favoured in the presence of the salt, so interaction with the macromolecule is at low affinity sites. PMID- 11496837 TI - Alcalase rapeseed inhibitors: purification and partial characterization. AB - Extensive rapeseed protein hydrolysate obtained sequentially with Alcalase and Flavourzyme showed inhibitory activity towards Alcalase. Inhibitory activity decreased as the hydrolytic process progressed probably by heat denaturation and/or partial protease degradation. Alcalase rapeseed inhibitors were purified by gel filtration and subsequent ion exchange chromatography. They are composed of peptides of 8.4 and 6.1 kDa linked by interchain disulphide bonds, as observed by reducing SDS-PAGE, with a native molecular weight of 18 kDa. Aminoacid composition of the inhibitors was characterized by the high proportion of methionine (4.2%) and cysteine (4.6%). Alcalase inhibitors were partially resistant to heat treatment; after heating at 70 degrees C for 45 minutes more than 50% of the original inhibitory activity remained in the purified protein but after heating at 90 degrees C for 5 minutes, inhibitory activity decreased very fast to a basal level. The possible relation of these protease inhibitors with the 2S albumin storage proteins is discussed. PMID- 11496838 TI - Vein graft disease:avoiding problems. PMID- 11496839 TI - Computer aided detection. PMID- 11496840 TI - 2001 mammography and CAD survey. PMID- 11496841 TI - Reviews should be more systematic. PMID- 11496842 TI - Defining the profession--talkin' 'bout your generation: how age diversity is changing practices. PMID- 11496843 TI - Why are you still posting manually? PMID- 11496844 TI - [Diagnosis of pancreas annulare using MR-cholangiopancreatography]. PMID- 11496845 TI - Summaries for patients. Whether albumin therapy improves or worsens survival of critically ill patients is not known. PMID- 11496846 TI - Summaries for patients. Improving protection against infection in people without enough immunoglobulin. PMID- 11496847 TI - Summaires for patients. Factors associated with the development of antibiotic resistant bacteria. PMID- 11496848 TI - Summaries for patients. Helicobacter pylori infection and the risk for heart attacks in men. PMID- 11496849 TI - Resumption of driving after life-threatening ventricular tachyarrhythmia. AB - BACKGROUND: Although the privilege of driving must be respected, it may be necessary to restrict driving when it poses a threat to others. The risks associated with allowing patients with life-threatening ventricular tachyarrhythmias to drive have not been quantified. METHODS: The Antiarrhythmics versus Implantable Defibrillators (AVID) trial compared antiarrhythmic-drug therapy with the implantation of defibrillators in patients resuscitated from near-fatal ventricular arrhythmias. In the current study, we sent patients who participated in the AVID trial a questionnaire, to be completed anonymously, requesting information about driving habits and experiences. RESULTS: The questionnaire was returned by 758 of 909 patients (83 percent). Of these, 627 patients drove during the year before their index episode of ventricular tachyarrhythmia. A total of 57 percent of these patients resumed driving within 3 months after randomization in the AVID trial, 78 percent within 6 months, and 88 percent within 12 months. While driving, 2 percent had a syncopal episode, 11 percent had dizziness or palpitations that necessitated stopping the vehicle, 22 percent had dizziness or palpitations that did not necessitate stopping the vehicle, and 8 percent of the 295 patients with an implantable cardioverter defibrillator received a shock. Fifty patients reported having at least 1 accident, for a total of 55 accidents during 1619 patient-years of follow-up after the resumption of driving (3.4 percent per patient-year). Only 11 percent of these accidents were preceded by symptoms of possible arrhythmia (0.4 percent per patient-year). CONCLUSIONS: Most patients with ventricular tachyarrhythmias resume driving early. Although it is common for them to have symptoms of possible arrhythmia while driving, accidents are uncommon and occur with a frequency that is lower than the annual accident rate of 7.1 percent in the general driving population of the United States. PMID- 11496850 TI - Nelfinavir, efavirenz, or both after the failure of nucleoside treatment of HIV infection. AB - BACKGROUND: The optimal antiretroviral treatment for patients who have human immunodeficiency virus (HIV) viremia despite treatment with nucleoside reverse transcriptase inhibitors (nucleoside analogues) remains uncertain. We studied treatment with regimens that combined two nucleoside analogues, at least one of which was new, with the protease inhibitor nelfinavir, the nonnucleoside reverse transcriptase inhibitor efavirenz, or both. METHODS: The study included 195 patients who had been treated with nucleoside analogues only, and had a plasma HIV type 1 (HIV-1) RNA level of at least 500 copies per milliliter. Patients were randomly assigned to receive, in addition to two nucleoside analogues, nelfinavir, efavirenz, or nelfinavir plus efavirenz. The primary end point was a plasma HIV-1 RNA level of less than 500 copies per milliliter at week 16. A secondary end point was the composite of the HIV-1 RNA levels measured at weeks 40 and 48. RESULTS: At week 16 and at weeks 40 and 48, the proportions of patients in whom a plasma HIV-1 RNA level of less than 500 copies per milliliter was achieved were, respectively, 81 percent and 74 percent in the nelfinavir-plus efavirenz group, 69 percent and 60 percent in the efavirenz group, and 64 percent and 35 percent in the nelfinavir group. Quadruple therapy resulted in a higher rate of viral suppression in both the short term (P=0.03) and the long term (P=0.001) than did triple therapy with nelfinavir. Triple therapy with efavirenz conferred a higher rate of long-term suppression than triple therapy with nelfinavir (P=0.004). Quadruple therapy also achieved a higher rate of virologic suppression than triple therapy with efavirenz (P=0.008). CONCLUSIONS: In HIV infected patients previously treated with nucleoside analogues, treatment with nelfinavir plus efavirenz and at least one new nucleoside analogue achieves a higher rate of viral suppression than do regimens with nucleoside analogues and nelfinavir or efavirenz alone. PMID- 11496851 TI - Dysfunction of endothelial protein C activation in severe meningococcal sepsis. AB - BACKGROUND: Impairment of the protein C anticoagulation pathway is critical to the thrombosis associated with sepsis and to the development of purpura fulminans in meningococcemia. We studied the expression of thrombomodulin and the endothelial protein C receptor in the dermal microvasculature of children with severe meningococcemia and purpuric or petechial lesions. METHODS: We assessed the integrity of the endothelium and the expression of thrombomodulin and the endothelial protein C receptor in biopsy specimens of purpuric lesions from 21 children with meningococcal sepsis (median age, 41 months), as compared with control skin-biopsy specimens. RESULTS: The expression of endothelial thrombomodulin and of the endothelial protein C receptor was lower in the patients with meningococcal sepsis than in the controls, both in vessels with thrombosis and in vessels without thrombosis. On electron microscopical examination, the endothelial cells were generally intact in both thrombosed and nonthrombosed vessels. Plasma thrombomodulin levels in the children with meningococcal sepsis (median, 6.4 ng per liter) were higher than those in the controls (median, 3.6 ng per liter; P=0.002). Plasma levels, protein C antigen, protein S antigen, and antithrombin antigen were lower than those in the controls. In two patients treated with unactivated protein C concentrate, activated protein C was undetectable at the time of admission, and plasma levels remained low. CONCLUSIONS: In severe meningococcal sepsis, protein C activation is impaired, a finding consistent with down-regulation of the endothelial thrombomodulin-endothelial protein C receptor pathway. PMID- 11496852 TI - Cerebral sinovenous thrombosis in children. AB - BACKGROUND: Cerebral sinovenous thrombosis in children is a serious disorder, and information is needed about its prevention and treatment. METHODS: The Canadian Pediatric Ischemic Stroke Registry was initiated in 1992 at the 16 pediatric tertiary care centers in Canada. Children (newborn to 18 years of age) with symptoms and radiographic confirmation of sinovenous thrombosis were included. RESULTS: During the first six years of the registry, 160 consecutive children with sinovenous thrombosis were enrolled, and the incidence of the disorder was 0.67 cases per 100,000 children per year. Neonates were most commonly affected. Fifty-eight percent of the children had seizures, 76 percent had diffuse neurologic signs, and 42 percent had focal neurologic signs. Risk factors included head and neck disorders (in 29 percent), acute systemic illnesses (in 54 percent), chronic systemic diseases (in 36 percent), and prothrombotic states (in 41 percent). Venous infarcts occurred in 41 percent of the children. Fifty-three percent of the children received antithrombotic agents. Neurologic deficits were present in 38 percent of the children, and 8 percent died; half the deaths were due to sinovenous thrombosis. Predictors of adverse neurologic outcomes were seizures at presentation and venous infarcts. CONCLUSIONS: Sinovenous thrombosis in children affects primarily neonates and results in neurologic impairment or death in approximately half the cases. The occurrence of venous infarcts or seizures portends a poor outcome. PMID- 11496853 TI - Images in clinical medicine. Mesothelioma. PMID- 11496854 TI - Toxic leukoencephalopathy. PMID- 11496855 TI - The coxibs, selective inhibitors of cyclooxygenase-2. PMID- 11496856 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 24-2001. A 46-year-old woman with chronic sinusitis, pulmonary nodules, and hemoptysis. PMID- 11496857 TI - Driving after ventricular arrhythmias. PMID- 11496858 TI - Antiretroviral therapy for previously treated patients. PMID- 11496859 TI - Learn nothing, forget nothing--the Medicare Commission redux. PMID- 11496860 TI - Medicare reform--now is the time. PMID- 11496861 TI - The effectiveness of the varicella vaccine. PMID- 11496862 TI - Effect of early or delayed tympanostomy-tube insertion for persistent otitis media. PMID- 11496863 TI - Effect of early or delayed tympanostomy-tube insertion for persistent otitis media. PMID- 11496864 TI - Fluvoxamine for the treatment of anxiety disorders in children and adolescents. PMID- 11496865 TI - Spontaneous dissection of the carotid and vertebral arteries. PMID- 11496866 TI - Spontaneous dissection of the carotid and vertebral arteries. PMID- 11496867 TI - Epilepsy. PMID- 11496868 TI - Epilepsy. PMID- 11496869 TI - "Invisible" synthetic opiates and acute psychosis. PMID- 11496870 TI - Web sites with misinformation about illicit drugs. PMID- 11496871 TI - The link between health and economic security for working-age women. PMID- 11496872 TI - Plight of elderly people who are made homeless in hospital. PMID- 11496873 TI - Health needs of asylum seekers and refugees. Specific treatments are effective in cases of post-traumatic stress order. PMID- 11496874 TI - Health needs of asylum seekers and refugees. Head injury needs to be taken into consideration in survivors of torture. PMID- 11496875 TI - Violence towards female prostitutes. Violence in sex work extends to more than risks from clients. PMID- 11496876 TI - Violence towards female prostitutes. We should listen to working women. PMID- 11496877 TI - Management of prisoners with HIV infection. Prevention would be better than care. PMID- 11496878 TI - Management of prisoners with HIV infection. Real commitment to prevention is needed. PMID- 11496879 TI - Physical health of people with severe mental illness. Adequate staffing and shared commitment are needed. PMID- 11496880 TI - Risk of suicide related to income level in mental illness. Psychiatric disorders are more severe amount suicide victims of higher occupational level. PMID- 11496881 TI - Physical health of people with severe mental illness. General practitioners play a valuable part. PMID- 11496882 TI - Risk of suicide related to income level in mental illness. Direct association between social status and risk of suicide was not found in Germany. PMID- 11496883 TI - Neomycin should not be used to treat hepatic encephalopathy. PMID- 11496884 TI - Long term anticoagulation or antiplatelet treatment. Inclusion criteria determine results of review. PMID- 11496886 TI - Long term anticoagulation or antiplatelet treatment. Patients at risk of stroke should be given warfarin. PMID- 11496885 TI - Long term anticoagulation or antiplatelet treatment. Only warfarin has been shown to reduce stroke risk in patients with atrial fibrillation. PMID- 11496887 TI - Long term anticoagulation or antiplatelet treatment. Giving warfarin always depends on balancing risks. PMID- 11496888 TI - Long term anticoagulation or antiplatelet treatment. Garbage in equals garbage out. PMID- 11496889 TI - Long term anticoagulation or antiplatelet treatment. Drug name was incorrect. PMID- 11496890 TI - Long term anticoagulation or antiplatelet treatment. How do we decide between warfarin and aspirin? PMID- 11496891 TI - Distressed women take contact tracing seriously. PMID- 11496892 TI - Communicable disease control must remain at local level. PMID- 11496893 TI - Reviewing screening mammograms with newly diagnosed patients is another unnecessary burden. PMID- 11496898 TI - HCFA (now CMS) issues ruling on coincidence imaging. PMID- 11496899 TI - Highlights 2001 Lecture: against all odds, nuclear medicine has thrived. PMID- 11496900 TI - Diagnostics and treatment of HIV-discordant couples who wish to have children. AB - Over the last years the reality surrounding HIV-infection has undergone a considerable change with regard to the life expectancy of patients, and the plans they can make for their lives. Because the majority of the HIV positive population is of reproductive age, one of these plans might be to have children, often as an expression of a fulfilled partnership. The need for medical support to realize this wish, however, is often confronted with ethical, medical or forensic restraints. For this reason, interdisciplinary recommendations have now been developed--for the first time on a global basis--which aim to provide guidelines for practitioners in this complicated area. PMID- 11496901 TI - Science lobbying tactics. PMID- 11496902 TI - Dietary fat: at the heart of the matter. PMID- 11496903 TI - Dietary fat: at the heart of the matter. PMID- 11496904 TI - Dietary fat: at the heart of the matter. PMID- 11496905 TI - Importance of careful patient selection for LASIK. PMID- 11496906 TI - Possible case of capsular block syndrome. PMID- 11496907 TI - Macular hemorrhage after LASIK for high myopia: a causal association? PMID- 11496908 TI - Meaningful representation of power. PMID- 11496909 TI - Cataract surgery in the nanophthalmic eye. PMID- 11496910 TI - Single injection of anesthetic agent. PMID- 11496911 TI - Trypan blue mixed with sodium hyaluronate for capsulorhexis. PMID- 11496912 TI - Independent study of excimer laser ablation of the cornea. PMID- 11496913 TI - Intravitreal antibiotics: the emergency kit. PMID- 11496914 TI - Consultation section: refractive surgical problem. PMID- 11496915 TI - Polio eradication: final 1% poses greatest challenge. PMID- 11496916 TI - As burden of mental disorders looms large, countries report lack of mental health programmes. PMID- 11496917 TI - The optimal duration of exclusive breastfeeding. PMID- 11496918 TI - Eye clinics leave blind and partially sighted patients abandoned and confused. PMID- 11496920 TI - When treating patients with schizophrenia, what clinical points should be considered if lamotrigine is chosen to augment clozapine? PMID- 11496919 TI - Clinical picture: eyelid metastasis. PMID- 11496921 TI - The spinal cord research foundation: two decades of progress. AB - Over the course of the past 24 years, the Paralyzed Veterans of America's Spinal Cord Research Foundation (SCRF) has provided support for more than 400 research grants in a wide range of areas, from improved wheelchair design to axon pathfinding in Drosophila. The Founders of SCRF, as well as its current trustees, believe that it is imperative to target a broad range of research areas to maximize the quality of life for people, both veterans and nonveterans, with paralysis. This approach has involved the support of basic science and clinical research directed towards repair of the spinal cord, as well as research into improved treatments for complications of spinal cord dysfunction and other projects, including engineering grants and conferences, that may enhance the quality of life for people with paralysis within the immediate future. PMID- 11496923 TI - NEURON: a tool for neuroscientists. AB - NEURON is a simulation environment for models of individual neurons and networks of neurons that are closely linked to experimental data. NEURON provides tools for conveniently constructing, exercising, and managing models, so that special expertise in numerical methods or programming is not required for its productive use. This article describes two tools that address the problem of how to achieve computational efficiency and accuracy. PMID- 11496922 TI - Glial tumor invasion: a role for the upregulation and cleavage of BEHAB/brevican. AB - Glial tumors, gliomas, are the most common primary intracranial tumors. Their distinct ability to invade the normal surrounding tissue makes them difficult to control and nearly impossible to completely remove surgically, and it accounts for the extraordinarily high lethality associated with gliomas. The ability of these transformed glial cells to invade the normal surrounding tissue is relatively unique in the adult CNS, which under most circumstances, is inhibitory to cell movement. The extracellular matrix (ECM) can modulate, in part, the permissiveness of a tissue to cell movement. Accordingly, the ability of gliomas to modify the ECM of the CNS may mediate the invasiveness of these cells. One ECM molecule that shows dramatic upregulation in gliomas is BEHAB (brain enriched hyaluronan binding)/brevican, a brain-specific chondroitin sulfate proteoglycan. BEHAB/brevican expression is also upregulated during periods of increased glial cell motility in development and following brain injury. Experimental evidence suggests that in glioma, in addition to upregulation of BEHAB/brevican, proteolytic processing of the full-length protein also may contribute to invasion. Here, the authors present a review of the literature on glial tumor invasion by modulation of the ECM and propose a two-step model for BEHAB/brevican's role in this process. PMID- 11496924 TI - Sodium channels and neurological disease: insights from Scn8a mutations in the mouse. AB - The human genome contains 10 voltage-gated sodium channel genes, 7 of which are expressed in neurons of the CNS and PNS. The availability of human genome sequences and high-throughput mutation screening methods make it likely that many human disease mutations will be identified in these genes in the near future. Mutations of Scn8a in the mouse demonstrate the broad spectrum of neurological disease that can result from different alleles of the same sodium channel gene. Null mutations of Scn8a produce motor neuron failure, loss of neuromuscular transmission, and lethal paralysis. Less severe mutations result in ataxia, tremor, muscle weakness, and dystonia. The effects of Scn8a mutations on channel properties have been studied in the Xenopus oocyte expression system and in neurons isolated from the mutant mice. The Scn8a mutations provide insight into the mode of inheritance, effect on neuronal sodium currents, and role of modifier genes in sodium channel disease, highlighting the ways in which mouse models of human mutations can be used in the future to understand the pathophysiology of human disease. PMID- 11496925 TI - Neuregulins and the shaping of synapses. AB - In developing and mature neural circuits, neural electrical activity controls the correct formation of connections and their state. Neuregulins (NRGs) mediate between the electrical neural activity and molecular components by regulating the expression of ion channel receptors or transmitter release in synapses. Furthermore, NRGs may be signaling factors involved in tuning locomotion or other higher functions by coordinating excitatory and inhibitory neurons. PMID- 11496926 TI - Nerve injury-induced pain in the trigeminal system. AB - This article reviews some recent findings on peripheral mechanisms related to the development of oro-facial pain after trigeminal nerve injury. Chronic injury induced oro-facial pain is not in itself a life-threatening condition, but patients suffering from this disorder undoubtedly have a reduced quality of life. The vast majority of the work on pain mechanisms has been carried out in spinal nerve systems. Those studies have provided great insight into mechanisms of neuropathic spinal pain, and much of the data from them is obviously relevant to studies of trigeminal pain. However, it is now clear that the pathophysiology of the trigeminal nerve (a cranial nerve) is in many ways different to that found in spinal nerves. Whereas some of the changes seen in animal models of trigeminal nerve injury mimic those occurring after spinal nerve injury (e.g., the development of spontaneous activity from the damaged axons), others are different, such as the time-course of the spontaneous activity, some of the neuropeptide changes in the trigeminal ganglion, and the lack of sprouting of sympathetic terminals in the ganglion. Recent findings provide new insights that help our understanding of the etiology of chronic injury-induced oro-facial pain. Future investigations will hopefully explain how data gained from these studies relate to clinical pain experience in man and should enable the rapid development of new therapeutic regimes. PMID- 11496927 TI - Maxi-K potassium channels: form, function, and modulation of a class of endogenous regulators of intracellular calcium. AB - Large-conductance calcium-activated (maxi-K, BK) potassium channels are widely distributed in the brain. Maxi-K channels function as neuronal calcium sensors and contribute to the control of cellular excitability and the regulation of neurotransmitter release. Little is currently known of any significant role of maxi-K channels in the genesis of neurological disease. Recent advances in the molecular biology and pharmacology of these channels have revealed sources of phenotypic variability and demonstrated that they can be successfully modulated by pharmacological agents. A potential role is suggested in the treatment of conditions such as ischemic stroke and cognitive disorders. PMID- 11496928 TI - Gait and balance dysfunction: a model of the interaction of age and disease. AB - This article reviews age-associated changes in gait, balance, and sensorimotor function and contrasts them to the changes that occur as a result of disease. Strength peaks in the mid-20s but declines only modestly until the fifth decade, after which there is rapid decline. All aspects of sensory function diminish with age, resulting in modest sensory changes in older patients. Gait speed is stable until the seventh decade and thereafter slows modestly. Age-related changes in the balance of older persons result in an effective response that meets routine needs but may be ineffective under demanding circumstances capable of producing response. Thus, these modest age-related decrements may be an element in the increased incidence of falls in older people. The loss of function caused by disease is of greater magnitude as well as being superimposed on that caused by age and thus may lead to a failure of mobility. Neurologic diseases that impair sensorimotor function compromise mobility in a manner consistent with the deficits that they produce. In mobility dysfunction caused by white matter lesions, the gait/balance abnormalities may not have unique clinical features, making diagnosis difficult. We used quantitative MRI to study the brains of older subjects with mobility impairment. The volume of white matter lesions in mobility impaired subjects was double that of controls and was unrelated to increasing age, suggesting that white matter lesions are a disease-related rather than an age-related occurrence. We also used stance perturbation evoked potentials to demonstrate delayed conduction in some mobility-impaired older subjects. Prolonged sensory conduction may contribute to maladaptive balance and thus may be useful for diagnosis. Contemporary imaging and neurophysiology thus provides insight into the pathophysiology of mobility impairment and allows for more accurate diagnosis. PMID- 11496929 TI - Thomas Jessell, who defined key mechanisms controlling development and functional organization of the spinal cord, wins Bristol-Myers Squibb Neuroscience Award. PMID- 11496930 TI - Fast kinetic analysis of ligand-gated ion channels. AB - Ligand-gated ion channels mediate fast synaptic transmission in the central and peripheral nervous system and the neuromuscular junction. Their common principle of function is the conversion of a chemical signal--neurotransmitter binding- into an electrical signal, i.e., an ion influx into the postsynaptic cell. The transient nature of this signal requires experimental setups that provide adequate temporal resolution and the use of transient kinetic analysis rather than equilibrium methods for a correct description of receptor function. Although the highly specialized geometry of a synapse that allows very rapid delivery of neurotransmitter is difficult to mimic in an experimental system, a variety of techniques for rapid kinetic analysis are available, making it possible to determine at least some steps of receptor function with sufficient accuracy. This article provides an overview of strategies and methods of fast ligand application and kinetic analysis using whole-cell and single channel patch clamp. PMID- 11496931 TI - Fertility desires and intentions of HIV-positive men and women. AB - CONTEXT: HIV-positive men and women may have fertility desires and may intend to have children. The extent of these desires and intentions and how they may vary by individuals' social and demographic characteristics and health factors is not well understood. METHODS: Interviews were conducted from September through December 1998 with 1,421 HIV-infected adults who were part of the HIV Cost and Services Utilization Study, a nationally representative probability sample of 2,864 HIV-infected adults who were receiving medical care within the contiguous United States in early 1996. RESULTS: Overall, 28-29% of HIV-infected men and women receiving medical care in the United States desire children in the future. Among those desiring children, 69% of women and 59% of men actually expect to have one or more children in the future. The proportion of HIV-infected women desiring a child in the future is somewhat lower than the overall proportion of U.S. women who desire a child. The fertility desires of HIV-infected individuals do not always agree with those of their partners: As many as 20% of HIV-positive men who desire children have a partner who does not Generally, HIV-positive individuals who desire children are younger, have fewer children and report higher ratings of their physical functioning or overall health than their counterparts who do not desire children, yet desire for future childbearing is not related to measures of HIV progression. HIV-positive individuals who expect children are generally younger and less likely to be married than those who do not. Multivariate analyses indicate that black HIlV-positive individuals are more likely to expect children in the future than are others. While HIV-positive women who already have children are significantly less likely than others both to desire and to expect more births, partner's HIV status has mixed effects: Women whose partner's HIVstatus is known are significantly less likely to desire children but are significantly more likely to expect children in the future than are women whose partner's HIV status is unknown. Moreover, personal health status significantly affects women's desire for children in the future but not men's, while health status more strongly influences men's expectations to have children. CONCLUSIONS: The fact that many HIV-infected adults desire and expect to have children has important implications for the prevention of vertical and heterosexual transmission of HIV, the need for counseling to facilitate informed decision-making about childbearing and childrearing, and the future demand for social services for children born to infected parents. PMID- 11496932 TI - Reproductive and sexual health benefits in private health insurance plans in Washington State. AB - CONTEXT: Although unintended pregnancy and sexually transmitted diseases (STDs) are considerable problems in the United States, private health insurance plans are inconsistent in their coverage of reproductive and sexual health services needed to address these problems. METHODS: A survey administered to a market representative sample of 12 health insurance carriers in Washington State assessed benefit coverage for gynecologic services, maternity services, contraceptive services, pregnancy termination, infertility services, reproductive cancer screening, STD services, HIV and AIDS services, and sterilization, as well as for the existence of confidentiality policies. "Core" services in each category were defined based on U.S. Preventive Services Task Force and other recommendations. RESULTS: Of the 91 top-selling plans on which data were collected, 8% were indemnity plans, 14% were point-of-service plans, 21% were preferred-provider organization plans and 57% were health maintenance organization (HMO)-type products; they had a combined enrollment of 1.4 million individuals. Coverage of core services varied widely by type of plan. While a high proportion of plans covered core gynecologic, maternity, reproductive cancer screening, STD and HIV and AIDS services, nearly half of plans did not cover any kind of contraceptive method. Approximately 13% of female enrollees did not have core coverage for gyneco!ogic services, 19% for matemity services, 75% for contraception, 37% for sterilization and 53% for pregnancy termination; 98% of women and men were not covered for infertility treatment. Most carriers did not have specific policies for maintaining privacy of sensitive health information. Overall, benefit coverage was lower for indemnity, preferred-provider organization and HMO plans in Washington State than has previously been seen nationally. CONCLUSIONS: A sizable proportion of women and men in Washington State who rely on private-sector health insurance lack comprehensive coverage for key reproductive and sexual health services. PMID- 11496933 TI - Tubal sterilization in the United States, 1994-1996. AB - CONTEXT: Although the number and rate of tubal sterilizations, the settings in which they are performed and the characteristics of women obtaining sterilization procedures provide important information on contraceptive practice and trends in the United States, such data have not been collected and tabulated for manyyears. METHODS: Information on tubal sterilizations from the National Hospital Discharge Survey and the National Survey of Ambulatory Surgery was analyzed to estimate the number and characteristics of women having a tubal sterilization procedure in the United States during the period 1994-1996 and the resulting rates of tubal sterilization. These results were compared with those of previous studies to examine trends in clinical setting, in the timing of the procedure and in patient characteristics. RESULTS: In 1994-1996, more than two million tubal sterilizations were performed, for an average annual rate of 1 1.5 per 1,000 women; half were performed postpartum and half were interval procedures (i. e., were unrelated by timing to a pregnancy). All postpartum procedures were performed during inpatient hospital stays, while 96% of interval procedures were outpatient procedures. Postpartum sterilization rates were higher than interval sterilization rates among women 20-29 years of age; interval sterilization procedures were more common than postpartum procedures at ages 35-49. Sterilization rates were highest in the South. For postpartum procedures, private insurance was the expectedprimary source of payment for 48% and Medicaid was expected to pay for 41 %; for interval sterilization procedures, private insurance was the expected primary source of payment for 68% and Medicaid for 24%. CONCLUSIONS: Outpatient tubal sterilizations andprocedures using laparoscopy have increased substantially since the last comprehensive analysis of tubal sterilization in 1987, an indication of the effect of technical advances on the provision of this service. Continued surveillance of both inpatient and outpatient procedures is necessary to monitor the role of tubal sterilization in contraceptive practice. PMID- 11496935 TI - Increasing access to emergency contraception through community pharmacies: lessons from Washington State. PMID- 11496934 TI - States' implementation of the Section 510 abstinence education program, FY 1999. AB - CONTEXT: As part of its reworking of the nation's welfare system in 1996, Congress enacted a major new abstinence education initiative (Section 510 of Title V of the Social Security Act), projected to spend $87.5 million in federal, state and local funds per year for five years. The new program is designed to emphasize abstinence from sexual activity outside of marriage, at any age, rather than premarital abstinence for adolescents, which was typical of earlier efforts. The actual message and impact of the program, however, will depend on how it is implemented. METHODS: Program coordinators in all 50 states, the District of Columbia and Puerto Rico were surveyed concerning implementation of the Section 510 abstinence education program in FY 1999. The questionnaire asked about expenditures and activities performed, about policies established for a variety of specific situations and about how the term "sexual activity" is defined and what specific components of the federal definition of "abstinence education" are emphasized. RESULTS: Forty-five jurisdictions spent a total of $69 million through the Section 510 program in FY 1999. Of this total, $33 million was spent through public entities, $28 million was spent through private entities and $7million (in 22 jurisdictions) was spent through faith-based entities. Almost all jurisdictions reported funding school-related activities, with 38 reporting in-school instruction and presentations. Twenty-eight jurisdictions prohibited organizations from providing information about contraception (aside from failure rates), even at a client's request, while only six jurisdictions prohibited information about sexually transmitted diseases. Few reported having a policy or rendering guidance about providing services addressing sexual abuse, sexual orientation or existing pregnancy and parenthood. Only six respondents said they defined "sexual activity" for purposes of the program, and 16 reported focusing on specific portions of the federal definition of "abstinence education." CONCLUSIONS: More than one in 10 Section 510 dollars were spent through faith based entities. Programs commonly conducted in-school activities, particularly instruction and presentations, not only through public entities, but also through private and faith-based entities. Most jurisdictions prohibited the provision of information about contraception, about providers of contraceptive services or about both topics, even in response to a direct question and when using other sources of funding. Most also left definitions of "abstinence" and "sexual activity" as local decisions, thus not clearly articulating what the program is designed to encourage clients to abstain from. PMID- 11496936 TI - Using photographs to strengthen family planning research. PMID- 11496937 TI - Supersaturation: enhancement of skin penetration and permeation of a lipophilic drug. AB - PURPOSE: To increase the dermal delivery of a lipophilic model compound (LAP), and to deduce the underlying mechanism of enhanced absorption. METHODS: Penetration of LAP from mixtures of up to four degrees of saturation into the stratum corneum was evaluated using a tape-stripping method; epidermal permeation of the drug was measured in Franz diffusion cells. The relative diffusion and stratum corneum-vehicle partition coefficients of LAP were determined by fitting the results to the appropriate solutions to Fick's second law of diffusion. RESULTS: Both the skin permeation rate and the amount of LAP in the stratum corneum increased linearly with increasing degree of saturation. The apparent diffusivity and its partition coefficient deduced from the penetration experiments were independent of the degree of saturation of the drug in the applied formulation, and consistent with corresponding parameters derived from the permeation experiments. CONCLUSIONS: Supersaturation can increase the skin penetration and permeation of lipophilic drugs. The diffusion and partition parameters deduced for LAP indicate that supersaturation acts exclusively via increased thermodynamic activity without apparent effect on the barrier function of the skin per se. PMID- 11496938 TI - X-ray microanalysis of cryopreserved human skin to study the effect of iontophoresis on percutaneous ion transport. AB - PURPOSE: To study at the ultrastructural level which part of the skin is associated with percutaneous iodide transport by passive diffusion and iontophoresis. METHODS: Following passive diffusion or iontophoresis of iodide, the morphology and the ion distribution of the skin was preserved by rapid freezing. The skin was kept frozen until and during examination by transmission electron microscopy (TEM) and X-ray microanalysis (XRMA). The intrinsic electron absorbing characteristics of cryopreserved skin allow direct TEM examination without additional staining. XRMA can be used to obtain in a relatively nondestructive way in situ information on ion distributions across the skin. RESULTS: After passive diffusion, iodide was mainly found in the stratum corneum (SC), whereas there was little iodide in the viable epidermis. Iontophoresis up to 300 microA/cm2 did not significantly affect this distribution. With iontophoresis at 1,000 microA/cm2, the amount of iodide increased dramatically and was equally distributed over the SC and viable epidermis. The presence of iodide in the SC suggests that iodide is present inside corneocytes. CONCLUSIONS: Iontophoresis up to 300 microA/cm2 does not significantly perturb skin structures in contrast to iontophoresis at 1,000 microA/cm2. The presence of iodide inside corneocytes suggests the possibility of transcellular percutaneous iodide transport. PMID- 11496939 TI - Effect of bilayer distruption on transdermal transport of low-molecular weight hydrophobic solutes. AB - PURPOSE: Applications of transdermal drug delivery are limited by low skin permeability. Several chemicals have been used to enhance transdermal drug transport, many of which enhance skin permeability by disordering lipid bilayers. The objective of this study was to develop a mathematical model to describe the effect of bilayer disrupting agents on skin permeability to low molecular weight hydrophobic drugs. METHODS: I predicted solute partition and diffusion coefficients in the lipid bilayers of the stratum corneum using scaled particle theory, which calculates these coefficients based on the work required to create a cavity to incorporate the solute in the lipid bilayer. RESULTS: Model equations predicted that no significant permeability enhancement would be observed for small solutes (MW < 100). Thereafter, the enhancement, E, increases with solute cross-sectional area. The resulting equation to predict the enhancement of skin permeability is given by E = exp[alpha(r2 - 8.7)], where r is solute molecular radius in angstroms and alpha is the degree of bilayer disorder. Predictions of the model were compared with the experimental data collected from several studies. CONCLUSIONS: The model predictions compare well with the experimental data. PMID- 11496940 TI - Divergent effects of raloxifene HCI on the pharmacokinetics and pharmacodynamics of warfarin. AB - PURPOSE: Evista (raloxifene HCl) is a nonsteroidal selective estrogen receptor modulator that displays estrogen agonist effects on bone and lipid metabolism but estrogen antagonist effects on the breast and endometrium. The potential for drug drug interaction between raloxifene and warfarin was assessed in 15 healthy postmenopausal women. METHODS: Single doses of warfarin (20 mg) were administered prior to and during 2 weeks of dosing with raloxifene 120 mg/day. Each warfarin dose was followed by pharmacokinetic sampling and prothrombin time measurements. RESULTS: Raloxifene administration resulted in 7.1% and 14.1% decreases in the clearance (CLp/F) and 7.4% and 9.8% decreases in the volume of distribution (Vss/F) of R- and S-warfarin, respectively (all p < or = 0.05). In contrast to the slightly higher plasma concentrations of R- and S-warfarin, raloxifene reduced the maximum prothrombin time (PTmax) by 10% and the area under the PT versus time curve from 0-120 h (AUCPT) by 8% (p < 0.01). CONCLUSIONS: Raloxifene administration may result in a small increase in systemic warfarin exposure that is associated with a diminution, not augmentation, of the pharmacodynamic effect. Due to the small magnitude of this effect, concomitant administration of raloxifene and warfarin is not likely to result in clinically significant drug drug interaction. PMID- 11496941 TI - Pharmacokinetics of the R- and S-enantiomers of oxybutynin and N desethyloxybutynin following oral and transdermal administration of the racemate in healthy volunteers. AB - PURPOSE: To characterize the enantiomers of oxybutynin (OXY) and N desethyloxybutynin (DEO) following transdermal and oral administration. METHODS: OXY was administered either as a single transdermal system over a 96 h wear period or as a single 5 mg immediate-release tablet to 18 healthy male and female subjects in a randomized, open-label, two-way crossover design. Blood samples were collected for 108 h after application of the transdermal system and for 6 h after oral administration. Plasma concentrations of the R- and S-enantiomers of OXY and DEO were assayed by LC-MS/MS. Enantiomer in vitro skin flux was evaluated using human cadaver skin. RESULTS: In vitro skin flux studies demonstrated equal absorption of R and S- OXY. Plasma concentrations and pharmacokinetic parameters of the R-enantiomers of OXY and DEO were slightly lower than the S-enantiomers following transdermal OXY. The relative AUC values were S-OXY>S-DEO>R-OXY>R-DEO. The AUC ratios of DEO/ OXY were less than 1 for both the R- and S- enantiomers. Following oral dosing, plasma DEO concentrations greatly exceeded OXY resulting in relative AUC values of R-DEO>S-DEO>S-OXY>R-OXY. The mean AUC ratios of S- and R-DEO/OXY were 3.25 and 8.93, respectively. CONCLUSIONS: Stereoselective metabolism of OXY was evident following both transdermal and oral administration of OXY. The reduced pre-systemic metabolism of transdermally administered OXY compared to oral administration resulted in not only significantly lower DEO plasma concentrations, but also a different metabolite pattern. The differences between R-OXY and R-DEO following the two routes of administration support the potential for comparable clinical efficacy and reduced anticholinergic side effects with transdermal treatment. PMID- 11496942 TI - Cisplatin-loaded polymer-metal complex micelle with time-modulated decaying property as a novel drug delivery system. AB - PURPOSE: The pharmacological activity and pharmacokinetics of cisplatin (CDDP) loaded polymeric micelles were examined to reveal their usefulness as a novel tumor-directed drug carrier system of CDDP. METHODS: In biodistribution assay, free CDDP or CDDP-loaded micelles were administered intravenously to Lewis lung carcinoma-bearing mice. Antitumor activity and nephrotoxicity were respectively evaluated by the measurement of tumor size and plasma blood urea nitrogen (BUN) after single bolus i.v. administration of each drug. RESULTS: The time profile of the plasma Pt level after the injection of the micelles exhibited a time modulated disappearance as observed in saline in vitro. The micelles exhibited 5.2- and 4.6-fold higher AUC of Pt in the plasma and tumor, respectively, with minimal change in the kidney, in comparison with free CDDP, suggesting that prolonged circulation of Pt in circulation and specific accumulation in the tumor were achieved utilizing the micellar drug carrier system. Administration of the micelles at the dose exhibiting antitumor activity similar to free CDDP did not increase the plasma BUN, whereas free CDDP induced its remarkable increase. CONCLUSION: CDDP-loaded micelles restrained nephrotoxicity, which is the dose limiting factor of CDDP, while exhibiting tumor-specific accumulation. Thus, CDDP loaded micelles are expected to be a novel formulation of CDDP for clinical use. PMID- 11496943 TI - Liposomes bearing polyethyleneglycol-coupled transferrin with intracellular targeting property to the solid tumors in vivo. AB - PURPOSE: The purpose of this study was to determine the usefulness of transferrin (TF)-pendant-type polyethyleneglycol (PEG)-liposomes (TF-PEG-liposomes), in which TF was covalently linked to the distal terminal of PEG chains on the external surface of PEG-liposomes as a carrier for in vivo cytoplasmic targeting to tumor cells. METHODS: Small unilamellar TF-PEG-liposomes (100-140 nm in diameter) were prepared from DSPC, CH, DSPE-PEG, and DSPE-PEG-COOH (2:1:0.11:0.021, molar ratio), and were conjugated to TF via the carboxyl residue of DSPE-PEG-COOH. The intracellular targeting ability of TF-PEG-liposomes to tumor cells was examined in vitro and in Colon 26 tumor-bearing mice. RESULTS: TF-PEG-liposomes, bearing approximately 25 TF molecules per liposome, readily bound to mouse Colon 26 cells in vitro and were internalized by receptor-mediated endocytosis. TF-PEG-liposomes showed a prolonged residence time in the circulation and low RES uptake in Colon 26 tumor-bearing mice, resulting in enhanced extravasation of the liposomes into the solid tumor tissue. Electron microscopic studies in Colon 26 tumor-bearing mice revealed that the extravasated TF-PEG-liposomes were internalized into tumor cells by receptor-mediated endocytosis. CONCLUSION: TF-PEG-liposomes had the capabilities of specific receptor binding and receptor-mediated endocytosis to target cells after extravasation into solid tumors in vivo. Such liposomes should be useful for in vivo cytoplasmic targeting of chemotherapeutic agents or plasmid DNAs to target cells. PMID- 11496944 TI - Role of genetic algorithms and artificial neural networks in predicting the phase behavior of colloidal delivery systems. AB - PURPOSE: A genetic neural network (GNN) model was developed to predict the phase behavior of microemulsion (ME), lamellar liquid crystal (LC), and coarse emulsion forming systems (W/O EM and O/W EM) depending on the content of separate components in the system and cosurfactant nature. METHOD: Eight pseudoternary phase triangles, containing ethyl oleate as the oil component and a mixture of two nonionic surfactants and n-alcohol or 1,2-alkanediol as a cosurfactant, were constructed and used for training, testing, and validation purposes. A total of 21 molecular descriptors were calculated for each cosurfactant. A genetic algorithm was used to select important molecular descriptors, and a supervised artificial neural network with two hidden layers was used to correlate selected descriptors and the weight ratio of components in the system with the observed phase behavior. RESULTS: The results proved the dominant role of the chemical composition, hydrophile-lipophile balance, length of hydrocarbon chain, molecular volume, and hydrocarbon volume of cosurfactant. The best GNN model, with 14 inputs and two hidden layers with 14 and 9 neurons, predicted the phase behavior for a new set of cosurfactants with 82.2% accuracy for ME, 87.5% for LC, 83.3% for the O/W EM, and 91.5% for the W/O EM region. CONCLUSIONS: This type of methodology can be applied in the evaluation of the cosurfactants for pharmaceutical formulations to minimize experimental effort. PMID- 11496945 TI - Fractal volume of drug distribution: it scales proportionally to body mass. AB - PURPOSE: To develop the physiologically sound concept of fractal volume of drug distribution, vf, and evaluate its utility and applicability in interspecies pharmacokinetic scaling. METHODS: Estimates for vf of various drugs in different species were obtained from the relationship: vf = (v - Vpl)(Vap - Vpl)/V + Vpl where v is the total volume of the species (equivalent to its total mass assuming a uniform density Ig/mL), Vpl is the plasma volume of the species and Vap is the conventional volume of drug distribution. This equation was also used to calculate the fractal analogs of various volume terms of drug distribution (the volume of central compartment, Vc, the steady state volume of distribution, Vss, and the volume of distribution following pseudodistribution equilibrium, Vz). The calculated fractal volumes of drug distribution were correlated with body mass of different mammalian species and allometric exponents and coefficients were determined. RESULTS: The calculated values of vf for selected drugs in humans provided meaningful and physiologically sound estimates for the distribution of drugs in the human body. For all fractal volume terms utilized, the allometric exponents were found to be either one or close to unity. The estimates of the allometric coefficients were found to be in the interval (0,1). These decimal values correspond to a fixed fraction of the fractal volume term relative to body mass in each one of the species. CONCLUSIONS: Fractal volumes of drug distribution scale proportionally to mass. This confirms the theoretically expected relationship between volume and mass in mammalian species. PMID- 11496946 TI - Modeling heterogeneity of properties and random effects in drug dissolution. AB - PURPOSE: To investigate new models characterizing dissolution data obtained for heterogenous materials (model I) and under randomly time-varying conditions (model II). METHODS: In model I, the heterogeneity of the dissolving substance introduces variation of the fractional dissolution rate. In model II, the fractional dissolution rate evolves randomly, and thus the dissolution has the characteristics of a stochastic process. This situation is studied for the constant and time-dependent means of the dissolution rate. RESULTS: The time dynamics of the dissolved fraction is presented for model I. The standard characteristics of dissolution are derived under general conditions and for several examples. One of them is in accordance with a function found empirically (1). A duality between the time-dependency of the fractional dissolution rate and the heterogeneity of the substance is investigated. The mean and variance of the dissolved fraction are calculated for model II. A method for estimating the mean dissolution rate is proposed and illustrated using Monte-Carlo experiments. CONCLUSIONS: It follows from model I that the heterogeneity, with the same mean properties, slows down the dissolution with respect to the homogeneous case. The second approach permits predictions about the role of the stochastic fluctuations of the dissolution rate and to establish the boundaries for the dissolution profiles. PMID- 11496947 TI - Controlled local delivery of interleukin-2 by biodegradable polymers protects animals from experimental brain tumors and liver tumors. AB - PURPOSE: The purpose of our study was to develop an injectable polymeric system for the long-term localized delivery of bioactive interleukin-2 for antitumor immunotherapy. METHODS: IL-2 was encapsulated into gelatin and chondroitin-6 sulfate using an aqueous-based complex coacervation. CTLL-2 cells were used to measure the bioactivity of released IL-2 and radiolabeled IL-2 was used for release studies in the rat brain and mouse liver. Antitumor efficacy studies were carried out in primary (9L gliosarcoma) and metastatic (B16-F10 melanoma) brain tumor models in rats and mice, respectively, as well as a murine liver tumor model (CT26 carcinoma). Survivors of the metastatic brain tumor challenge were rechallenged with tumor in the opposite lobe of the brain to confirm that antitumor immunologic memory had developed. RESULTS: Bioactive IL-2 was released for over 2 weeks in vitro and in vivo IL-2 release showed significant IL-2 levels for up to 21 days. Polymeric IL-2 microspheres injected intratumorally were statistically more effective in protecting animals challenged with fatal tumor doses in the brain and the liver than placebo or autologous tumor cells genetically engineered to secrete IL-2. Immunologic memory was induced following IL-2 microsphere therapy in the B16-F10 brain tumor model that was capable of protecting 42% of animals from a subsequent intracranial tumor challenge, suggesting that tumor destruction was mediated by the immune system. CONCLUSIONS: Local IL-2 therapy using novel polymeric carriers. aimed at stimulating long lasting antitumor immunity, may provide an improved method of treating a variety of cancers. PMID- 11496948 TI - Mdr1 transfection causes enhanced apoptosis by paclitaxel: an effect independent of drug efflux function of P-glycoprotein. AB - PURPOSE: We previously reported that in patient tumors the expression of the mdrl p-glycoprotein (Pgp) resulted in a lower paclitaxel-induced inhibition of DNA precursor incorporation, but a higher apoptosis (Clin. Cancer Res. 4:2949-2955, 1998). The present study was to evaluate these findings in an experimental system where the Pgp effect can be studied without confounding factors such as the intra and inter-tumor heterogeneity associated with patient tumors. METHODS: To separate the effect of Pgp on intracellular paclitaxel accumulation from its effects on drug sensitivity, we compared the drug activity at various extracellular and intracellular drug concentrations using the human breast MCF7 tumor cells and its mdr1-transfected variant BC19 cells. RESULTS: Compared to MCF7 cells, BC19 cells showed a 9-fold higher Pgp level and >13-fold higher mdrl expression. Intracellular paclitaxel accumulation was 80-130% lower in BC19 cells when the extracellular concentrations were < or = 100 nM, but the difference was reduced to <15% differences at higher extracellular concentrations of > or = 1,000 nM. For the G2/M block effect MCF7 cells were 43-fold more sensitive than BC19 cells at equal extracellular concentration, and 3.5-fold more sensitive at comparable intracellular concentrations. On the contrary. BC19 cells were more sensitive to the apoptotic effect: BC19 cells showed equal or higher apoptosis compared to MCF7 cells at extracellular concentrations above 100 nM, and a 30 100% higher apoptosis at comparable intracellular concentrations. CONCLUSIONS: These results confirm our previous observations in patient tumors and indicate that enhanced Pgp expression is associated with enhanced sensitivity to the apoptotic effect of paclitaxel and reduced sensitivity to its G2/M block effect, via yet-unknown mechanisms that are unrelated to the effect of Pgp on intracellular drug accumulation. PMID- 11496949 TI - LDL induced association of anionic liposomes with cells and delivery of contents as shown by the increase in potency of liposome dependent drugs. AB - PURPOSE: To establish whether anionic liposomes interact with the low-density lipoprotein (LDL) receptor, to determine the role of lipoproteins in this interaction, and whether the association causes functional delivery of encapsulated drugs. METHODS: The cell lines used were CV1-P and CHO wild type, both of which express the LDL receptor, and CHOldlA7, which lacks the LDL receptor. Cellular association of encapsulated methotrexate and fluorescein, labeled phosphatidylethanolamine in the lipid bilayer, was measured. Potency of three liposome dependent drugs (N-phosphonacetyl-L-aspartic acid, fluoroorotic acid, and methotrexate-gamma-aspartate) was also measured by growth inhibition. RESULTS: Association of liposomes containing at least 75 mol egg phosphatidylglycerol (ePG)/100 mol phospholipid with cells grown in defined medium supplemented with 1.0 mg/ml LDL was up to 30-fold higher with CV1-P or CHO wild type cells than with CHOldlA7, and 5-fold higher than association in defined medium lacking LDL. The addition of LDL did not yield any elevation of cellular association of distearoylphosphatidylglycerol liposomes. Increased association was paralleled by a corresponding increase in potency of all three liposome dependent drugs tested. CONCLUSIONS: ePG liposomes interact with the LDL receptor in an LDL-dependent fashion, and the interaction results in the delivery of contents to cells. PMID- 11496950 TI - Protamine sulfate enhances the transduction efficiency of recombinant adeno associated virus-mediated gene delivery. AB - PURPOSE: The purpose of this study was to evaluate glucose responsiveness in HepG2 human hepatoma cells transduced by a recombinant adeno-associated virus (rAAV) vector containing the insulin gene promoter. and to investigate the effect of protamine sulfate on rAAV-mediated gene delivery. METHODS: Recombinant AAV vector, AAV.Ins.Luc.delta EGFP, was employed to transduce HepG2 hepatoma cells. Virus infection was carried out either in the absence or presence of protamine sulfate, followed by fluorescence microscopic examination, luciferase activity assay, and flow cytometric analysis. Electrokinetic measurements were carried out to determine the effect of protamine sulfate on zeta potential of the cells and the virus. RESULTS: Glucose-responsive luciferase gene expression was obtained in rAAV-transduced HepG2 cells. Addition of 5 microg/ml protamine reversed the zeta potential of the cells and the virus particles, leading to enhanced transgene expression in rAAV-transduced HepG2 cells. Enhancement of protamine sulfate on rAAV-mediated gene transfer was dose-dependent. Addition of more than 5 microg/ml protamine resulted in a reduction of infectability of the virus. CONCLUSIONS: Glucose responsiveness in the millimolar concentration range can be obtained in rAAV-transduced HepG2 cells. Protamine sulfate, up to 5 microg/ml, enhanced the rAAV transduction efficiency in HepG2 cells. The enhancement was correlated with zeta potential of the cells and the virus. PMID- 11496951 TI - Interactions between oligonucleotides and cationic polymers investigated by fluorescence correlation spectroscopy. AB - PURPOSE: To evaluate whether fluorescence correlation spectroscopy (FCS) can be used to characterize the complexation between oligonucleotides and cationic polymers. METHODS: The features of the complexes between rhodamine labeled oligonucleotides (Rh-ONs) and poly(2-dimethylamino)ethyl methacrylate (pDMAEMA), poly(ethylene glycol)-poly(ethyleneimine) (pEG-pEI), and diaminobutane-dendrimer (NH2)64 (DAB64) were characterized by light scattering, electrophoretic mobility, electrophoresis, and FCS. RESULTS: At low polymer/Rh-ON ratios, a decrease of the fluorescence of the Rh-ONs was observed on binding of the Rh-ONs to all cationic polymers. This was explained by the creation of "multimolecular complexes" in which the Rh-labels quench each other. The multimolecular complexes, which are highly fluorescent as they carry a number of Rh-ONs, resulted in high fluorescence peaks in the fluorescence fluctuation profile as measured by FCS. For pDMAEMA and DAB64, at higher polymer/Rh-ON ratios the fluorescence of the polyplexes increased, caused by the formation of "monomolecular complexes," which consist of only one Rh-ON per polymer. In the case of pEG-pEI, the fluorescence stayed constant when the polymer/Rh-ON ratio increased, so multimolecular polyplexes remained. FCS confirmed these results as the high fluorescence peaks disappeared in case of pDMAEMA/Rh-ON and DAB64/Rh-ON dispersions, but remained present for pEG-pEI/Rh-ON dispersions. CONCLUSIONS: FCS seems applicable for study of the interactions between ONs and different types of cationic polymers. PMID- 11496952 TI - The mucosal toxicity of different benzalkonium chloride analogues evaluated with an alternative test using slugs. AB - PURPOSE: The objective of this study was to evaluate the mucosal toxicity of different benzalkonium chloride (BAC) analogues using slugs as the alternative test organism. METHODS: The effect of different BAC analogues on the mucosal tissue of slugs was determined from the protein, lactate dehydrogenase, and alkaline phosphatase released from the foot mucosa after treatment. Additionally, mucus production and reduction in body weight of the slugs were measured. The eye irritation potency of the molecules was evaluated with the Bovine Corneal Opacity and Permeability (BCOP) assay. The antimicrobial activity of the different BAC analogues was also assessed. RESULTS: All BAC analogues induced severe damage to the mucosal epithelium of the slugs, and the irritation increased with decreasing alkyl chain length: BAC-C16 < BAC-C14 < BAC-C12 approximately BAC-mix. A similar ranking was obtained with the BCOP assay for eye irritation. The relative order of activities among the three BAC analogues was the same, i.e., BAC-C14 > or = BAC-C16 > BAC-C12. The BAC-C14 exhibited higher activity than the BAC-mix. CONCLUSIONS: The toxicity and activity of BAC analogues depend on the alkyl chain length. The use of BAC-C14 as a conservative agent in pharmaceutical preparations instead of the BAC-mix should be considered. PMID- 11496953 TI - A new colorimetric assay for studying and rapid screening of membrane penetration enhancers. AB - PURPOSE: This work aims to demonstrate a novel chemical assay for rapid screening and analysis of the mode of action of membrane interaction by penetration enhancers. METHODS: The new bio-mimetic membrane assembly, consisting of supramolecular aggregates of lipids and conjugated polydiacetylene, undergoes visible and quantifiable blue-red color transitions upon interaction with penetration enhancers. RESULTS: The new colorimetric model has been employed to examine various classes of penetration enhancers, including 1-dodecylhexahydro-2H azepin-2-one (Azone), oleic acid, propylene-glycol, menthol, ethoxyglycol diethyleneglycol-monoethyl-ether (Transcutol), polysorbate-polyethylenesorbitan monolaurate (Tween-20), and the drug 7-chloro-1-methyl-5-phenyl-3H-1,4 benzodiazepin-2-one (Diazepam). The assay enables to evaluate the validity of various observations and hypotheses proposed in previous studies regarding permeation enhancement activities. Our results suggest, for example. that propylene glycol (PG) by itself does not interfere with membranes, but rather exhibits synergistic effect in combination with other penetration enhancers. Similarly, our data demonstrate that Transcutol does not independently interact with membranes. The colorimetric system also indicates that interaction of penetration enhancers with membranes depend upon the lipid phase, as well as the self-assembly properties of the enhancer molecules. CONCLUSIONS: The new biomimetic model membrane system can be applied for rapid screening of the activities of penetration enhancers, and provides insight into the mechanisms of permeability of membrane-active compounds. PMID- 11496954 TI - Targeting the sodium-dependent multivitamin transporter (SMVT) for improving the oral absorption properties of a retro-inverso Tat nonapeptide. AB - PURPOSE: To investigate the potential for delivering large peptides orally by altering their absorptive transport pathways and improving intestinal permeability. The absorptive transport of retro-inverso (R.I.-) K-Tat9 and R.I. K(biotin)-Tat9, novel peptidic inhibitors of the Tat protein of HIV-1, and their interactions with human SMVT (hSMVT), a high affinity, low capacity transporter, were investigated using Caco-2 and transfected CHO cells. METHODS: Following synthesis on a PAL resin using Fmoc chemistry, the transport of R.I.-K-Tat9 (0.01 25 microM) and R.I.-K(biotin)-Tat9 (0.1-25 microM) was evaluated across Caco-2 cells. The transport and kinetics of biotin, biocytin and desthiobiotin (positive controls for SMVT) were also determined. Uptake of R.I.-K-Tat9 and R.I.K(biotin) Tat9 (both 0.1-10 microM) was determined in CHO/hSMVT and CHO/pSPORT (control) cells. RESULTS: The absorptive transport of R.I.-K-Tat9 was passive, low (Pm approximately 1 x 10(-6) cm/sec) and not concentration dependent. R.I.K(biotin) Tat9 permeability was 3.2-fold higher than R.I.-K-Tat9 demonstrating active (Ea = 9.1 kcal/mole), concentration dependent and saturable transport (Km = 3.3 microM). R.I.-K(biotin)-Tat9 uptake in CHO/hSMVT cells (Km = 1.0 microM) was - 500-fold greater than R.I.-K-Tat9 (at 10 microM). R.I.-K(biotin)-Tat9 transport in Caco-2 and CHO/hSMVT cells was significantly inhibited by known substrates of SMVT including biotin, biocytin, and desthiobiotin. Passive uptake of R.I. K(biotin)-Tat9 was significantly greater than R.I.-K-Tat9 uptake in CHO/pSPORT cells. CONCLUSIONS: These results demonstrate that the structural modification of R.I.-K-Tat9 to R.I.-K(biotin)-Tat9 altered its intestinal transport pathway resulting in a significant improvement in its absorptive permeability by enhancing nonspecific passive and carrier-mediated uptake by means of SMVT. The specific interactions between R.I.-K(biotin)-Tat9 and SMVT suggest that targeting approaches utilizing transporters such as SMVT may substantially improve the oral delivery of large peptides. PMID- 11496955 TI - Effect of mdr1a P-glycoprotein gene disruption, gender, and substrate concentration on brain uptake of selected compounds. AB - PURPOSE: This study assessed the influence of mdr1a P-glycoprotein (P-gp) gene disruption, gender and concentration on initial brain uptake clearance (Clup) of morphine, quinidine and verapamil. METHODS: Clup of radiolabeled substrates was determined in P-gp-competent and deficient [mdr1a(-/-)] mice by in situ brain perfusion. Brain:plasma distribution of substrates after i.v. administration was determined in both strains. RESULTS: Genetic disruption of mdr1a P-gp resulted in 1.3-, 6.6- and 14-fold increases in Clup for morphine, verapamil and quinidine, respectively. With the exception of small differences for verapamil, gender did not affect Clup. Saturable transport of verapamil and quinidine was observed only in P-gp-competent mice, with apparent IC50 values for efflux of 8.6 +/- 2.3 microM and 36 +/- 2 microM, respectively. Verapamil Clup was approximately 50% higher in mdr1a(+/-) vs. mdr1a(+/+) mice; no such difference was observed for quinidine. In P-gp-competent mice, uptake of verapamil and quinidine was unaffected by organic vehicles. Plasma decreased VER Clup to a greater extent in the presence of P-gp. The influence of P-gp in situ was lower than, but correlated with, the effect in vivo. CONCLUSIONS: P-gp decreases Clup of morphine, verapamil and quinidine in situ with little or no influence of gender, but this effect cannot fully account for the effects of P-gp in vivo. P-gp is the only saturable transport mechanism for verapamil and quinidine at the murine blood-brain barrier. The influence of protein binding on Clup may be enhanced by P-gp-mediated efflux. PMID- 11496956 TI - Oral delivery of biologically active parathyroid hormone. AB - PURPOSE: Parathyroid hormone (PTH), the only drug known to stimulate bone formation. is a peptide therapeutic indicated in the treatment of osteoporosis. Unfortunately, PTH is only effective when dosed by injection because it has no oral bioavailability. Herein we report the oral absorption of PTH in rats and monkeys facilitated by the novel delivery agent, N-[8-(2-hydroxy-4 methoxy)bensoyl]amino caprylic acid (4-MOAC). METHODS: 4-MOAC was selected from a group of 100 delivery agents based on in vitro chromotography studies and in vivo screening studies in rats. The PTH/4-MOAC combination was then tested in monkeys. The interaction of 4-MOAC and PTH was evaluated by nuclear magnetic resonance (NMR) spectroscopy. RESULTS: Monkeys were administered an aqueous solution containing 4-MOAC and PTH and mean peak serum PTH concentrations of about 3000 pg/mL were obtained. The relative bioavailability of oral PTH was 2.1% relative to subcutaneous administration. The biological activity of the orally-delivered PTH was further evaluated in a rat model of osteoporosis. These studies showed that the bone formed following oral PTH/4-MOAC administration was comparable to that formed following PTH injections. The 4-MOAC mediated absorption of PTH is hypothesized to be the result of a noncovalent interaction between 4-MOAC and PTH. The preliminary evaluation of this interaction by NMR is described. CONCLUSIONS: 4-MOAC facilitates the absorption of PTH following oral administration to both rats and monkeys. The orally-absorbed PTH is biologically active as demonstrated in a rat model of osteoporosis. PMID- 11496957 TI - Receptor-mediated targeting of spray-dried lipid particles coformulated with immunoglobulin and loaded with a prototype vaccine. AB - PURPOSE: Spray-dried lipid-based microparticles (SDLM) serve as a platform for delivery of a wide variety of compounds including peptides, proteins, and vaccines to the respiratory mucosa. In the present study, we assessed the impact of IgG-mediated targeting to phagocytic cells of inactivated influenza virus formulated in SDLM, on subsequent immune responses. METHODS: SDLM were produced containing inactivated influenza virus strain A/WSN/32/H1N1 (WSN), with or without IgG. Using phagocytic antigen presenting cells (APC) and a T cell hybridoma (TcH) line specific for a dominant influenza virus epitope, we compared the in vitro responses elicited by ligand-formulated (SDLM-IgG-WSN) and non ligand particles (SDLM-WSN). The effect of including the IgG ligand in the formulation was further characterized by measuring the immune responses of rodents vaccinated with SDLM. RESULTS: SDLM-IgG-WSN were internalized in an Fc receptor (FcR)-dependent manner by phagocytic APC that were then able to effectively present a dominant, class II-restricted epitope to specific T cells. While SDLM-WSN elicited a lower response than administration of plain inactivated virus in saline, the level of the T cell response was restored both in vitro and in vivo by incorporating the APC FcR ligand, IgG, in the SDLM. CONCLUSIONS: Incorporation of FcR ligand (IgG) in SDLM restored the limited ability of formulated virus to elicit T-cell immunity, by receptor-mediated targeting to phagocytes. PMID- 11496959 TI - Searching for balanced hybrid NO-donor 1,4-dihydropyridines with basic properties. AB - PURPOSE: Model compounds containing NO-donor furoxan moieties at the 3-positioned basic lateral chain of 1, a 1,4-dihydropyridine related to nicardipine, were synthesized in order to study their vasodilating activity as well as their basic and lipophilic behaviour. METHODS: All the compounds were obtained by a modified Hantzsch approach. Potentiometry was used to determine pKa and lipophilicity descriptors. The furoxan 4-aryl-1,4-dihydropyridines were assessed for their ability to release nitrite, in the presence of a large excess of cysteine, by the Griess reaction. Vasodilating activity of the products in the absence and in the presence of ODQ, a well-known guanylate cyclase inhibitor, was evaluated on rat thoracic aorta. RESULTS: The compounds display low basicity values and for this reason their log Ds at physiological pH are identical to the log Ps of the neutral forms. Products 2, 3 display vasodilating action principally dependent on their Ca2+-antagonist properties, whereas 4 behaves as a well-balanced hybrid with mixed Ca2+-channel blocker and NO-dependent vasodilator activities. CONCLUSIONS. Nitrogen containing lateral chain at the 3-position of 1 is a suitable molecular region to be modified in order to obtain well-balanced furoxan NO-donor 1,4-DHPs. This manipulation produces a decrease in the basicity. General analysis of pKa and lipophilicity descriptors of these new DHPs suggest that molecular flexibility could influence both their basicity and log PI. PMID- 11496958 TI - Comparison of tissue concentrations after intramuscular and topical administration of ketoprofen. AB - PURPOSE: To assess whether topical ketoprofen, which has been reported to provide analgesic effects in clinical studies, reaches predictable tissue concentrations high enough to account for the reported analgesia. Intramuscular ketoprofen was used as positive control. METHODS: Muscle and subcutaneous tissue concentrations were assessed by microdialysis. Plasma and tissue concentrations after intramuscular injection were described using a three-compartment population pharmacokinetic model. The prediction performance of the model was assessed by superimposing tissue concentrations of 12 subjects that did not participate in the present study. RESULTS: Most dialysate concentrations after topical dosing of ketoprofen (100 mg) were below the quantification limit of 0.47 ng/ml. Plasma concentrations increased slowly and reached an apparent plateau of 7-40 ng/ml at 10-12h. No decline was observed up to 16 h. Tissue concentrations after intramuscular injection (100 mg) were about 10 times higher than those after topical dosing. Tissue concentrations measured in the majority of the 12 subjects that did not participate in the present study were found within the range of two thirds of the predicted concentrations. CONCLUSION: Predictable and cyclooxygenase-inhibiting concentrations of ketoprofen were achieved in subcutaneous and muscle tissue after intramuscular but not after topical dosing. Thus, the tissue concentrations of ketoprofen after topical administration can hardly explain the reported clinical efficacy of topical ketoprofen. PMID- 11496960 TI - Effect of selective lipid extraction from different body regions on epidermal barrier function. AB - PURPOSE: To assess the effects of selective lipid extraction and tape stripping on transepidermal water loss (TEWL) at three body regions in the pig. METHODS: Lipids were extracted from the abdominal, inguinal. and back regions using three different solvent extraction procedures or cellophane tape stripping (15x) on Yorkshire pigs. Three solvent extraction stages were I, cyclohexane (5 ml for three, 1-min extractions): II, cyclohexane/ethanol (4:1) (5 ml for three, 1-min extractions): and III, cyclohexane/ethanol (1:4) (5 ml for three, 3-min extractions) extracted as follows: Site A, Stage I: Site B, Stage I and II; Site C, Stage I, II and III. Erythema, edema, and TEWL were assessed in control, tape stripped, and extracted sites at 0, 6, and 24 h. The extracted lipids were analyzed by thin layer chromatography and quantified by densitometry for ceramide, cholesterol, cholesterol esters, fatty acids, and triglycerides. RESULTS: The change in TEWL (delta TEWL) in 14 of the 15 sites was the highest at 24 h and generally increased with each additional extraction. The greatest changes were present in the back. Each extraction stage removed specific lipids in reproducible quantities that caused the delta TEWL to increase from 0 to 24 h. Lipid removal was verified by transmission electron microscopy. The mean total lipid concentration depended on extraction solvents and body region, and was reproducible across sites and regions at equivalent stages of lipid extraction. Relative proportions of individual lipids extracted were similar across all body regions. Higher concentrations of total lipids were extracted from the back. CONCLUSIONS. These studies demonstrate that extraction of lipids increased the delta TEWL to a level similar to repeated tape stripping at all body sites in the pig. This study suggested that strategies that could biochemically alter epidermal lipid composition may increase absorption of simultaneously administered topical compounds and may be useful to enhance drug delivery. PMID- 11496961 TI - Probing the effect of vehicles on topical delivery: understanding the basic relationship between solvent and solute penetration using silicone membranes. AB - PURPOSE: In the present study we examined the relationship between solvent uptake into a model membrane (silicone) with the physical properties of the solvents (e.g., solubility parameter, melting point, molecular weight) and its potential predictability. We then assessed the subsequent topical penetration and retention kinetics of hydrocortisone from various solvents to define whether modifications to either solute diffusivity or partitioning were dominant in increasing permeability through solvent-modified membranes. METHODS: Membrane sorption of solvents was determined from weight differences following immersion in individual solvents, corrected for differences in density. Permeability and retention kinetics of 3H-hydrocortisone, applied as saturated solutions in the various solvents, were determined over 48 h in horizontal Franz-type glass diffusion cells. RESULTS: Solvent sorption into the membrane could be related to differences in solubility parameters, MW and hydrogen bonding (r2=0.76). The actual and predicted volume of solvent sorbed into the membrane was also found to be linearly related to Log hydrocortisone flux, with changes in both diffusivity and partitioning of hydrocortisone observed for the different solvent vehicles. CONCLUSIONS: A simple structure-based predictive model can be applied to the sorption of solvents into silicone membranes. Changes in solute diffusivity and partitioning appeared to contribute to the increased hydrocortisone flux observed with the various solvent vehicles. The application of this predictive model to the more complex skin membrane remains to be determined. PMID- 11496962 TI - Individual differences in alcohol-induced aggression. A nonhuman-primate model. AB - Some people are more likely than others to become aggressive after consuming alcohol. Researchers studying alcohol use and aggression hope to identify individual differences in behavior and biochemistry that exist among subjects who become aggressive following alcohol consumption. Research with nonhuman primates has shown that individual differences in brain chemistry predict impulsivity, aggression, and alcohol-induced aggression. These differences appear to be associated with early rearing experiences and remain stable throughout the individual's life. PMID- 11496963 TI - Victim and offender self-reports of alcohol involvement in crime. AB - Research suggests that a decreasing share of violent crime is attributable to offenders who had been drinking alcoholic beverages. Surveys of victims indicate that the rate of alcohol-involved violent crimes (i.e., crimes in which the perpetrators had been drinking, as perceived by the victims) decreased 34 percent from 1993 to 1998, whereas the rate of non-alcohol-involved violence decreased 22 percent. Surveys of some offenders also suggest that alcohol's role in violence is decreasing. The decrease in alcohol-involved violence is consistent with declines in other measures of alcohol use and misuse, including per capita alcohol consumption and alcohol involvement in traffic crashes. In contrast, violent offenders in State prisons are increasingly likely to report having used alcohol before committing their offenses, possibly illustrating the effect of more severe sanctions for alcohol-involved offenses. PMID- 11496964 TI - Court procedures for handling intoxicated drivers. AB - The courts have implemented numerous approaches to reduce the probability of recidivism among people apprehended for or convicted of driving while intoxicated. Although traditional punitive sanctions, such as fines and incarceration, are commonly used, they have not eliminated drinking and driving in the United States. Consequently, the court system has developed additional sanctioning procedures that show promise. For example, rehabilitative programs (e.g., alcohol education and alcoholism treatment) can reduce recidivism, at least marginally. These programs appear to be more effective when combined with license suspension. In addition to license suspension, several alternative methods for limiting driving opportunities of offenders have proven effective, including impounding offenders' vehicles or license plates, installing ignition interlocks, and requiring electronic home monitoring or house arrest. Effective court monitoring is a critical component in supporting recovery and compelling offenders to participate in rehabilitation programs. This role of the courts in monitoring offenders will likely increase as the use of intrusive, alternative sanctions grows. PMID- 11496965 TI - Alcohol and sexual assault. AB - Conservative estimates of sexual assault prevalence suggest that 25 percent of American women have experienced sexual assault, including rape. Approximately one half of those cases involve alcohol consumption by the perpetrator, victim, or both. Alcohol contributes to sexual assault through multiple pathways, often exacerbating existing risk factors. Beliefs about alcohol's effects on sexual and aggressive behavior, stereotypes about drinking women, and alcohol's effects on cognitive and motor skills contribute to alcohol-involved sexual assault. Despite advances in researchers' understanding of the relationships between alcohol consumption and sexual assault, many questions still need to be addressed in future studies. PMID- 11496966 TI - Antisocial personality disorder, alcohol, and aggression. AB - Epidemiologic studies and laboratory research consistently link alcohol use with aggression. Not all people, however, exhibit increased aggression under the influence of alcohol. Recent research suggests that people with antisocial personality disorder (ASPD) may be more prone to alcohol-related aggression than people without ASPD. As a group, people with ASPD have higher rates of alcohol dependence and more alcohol-related problems than people without ASPD. Likewise, in laboratory studies, people with ASPD show greater increases in aggressive behavior after consuming alcohol than people without ASPD. The association between ASPD and alcohol-related aggression may result from biological factors, such as ASPD-related impairments in the functions of certain brain chemicals (e.g., serotonin) or in the activities of higher reasoning, or "executive," brain regions. Alternatively, the association between ASPD and alcohol-related aggression may stem from some as yet undetermined factor(s) that increase the risk for aggression in general. PMID- 11496967 TI - Alcohol abuse as a risk factor for and consequence of child abuse. AB - The relationship between child abuse and the use or abuse of alcohol has two aspects. First, some findings have indicated that parental alcohol abuse may be associated with the physical or sexual abuse of children. Research findings in this area remain inconsistent, however. Second, the experience of being abused as a child may increase a person's risk for alcohol-related problems as an adult. This relationship has best been demonstrated in women who had been victims of childhood abuse. Several factors most likely contribute to or influence this relationship, including coping skills; antisocial behavior; and psychological problems, such as posttraumatic stress disorder. PMID- 11496968 TI - Alcohol-related intimate partner violence among white, black, and Hispanic couples in the United States. AB - Intimate partner violence (IPV) is a major public health problem in the United States. Results from a 1995 national study indicated that 23 percent of the black couples, 11.5 percent of the white couples, and 17 percent of the Hispanic couples surveyed reported an incident of male-to-female partner violence in the 12 months preceding the survey. The rate of female-to-male partner violence was also high: 15 percent among white couples, 30 percent among black couples, and 21 percent among Hispanic couples. The higher prevalence of IPV among ethnic minorities, compared with whites, cannot be explained by any single factor, but seems to be related to risk factors associated with the individual, the type of relationship between partners, and factors in the environment. Alcohol plays an important part in IPV. The study found that 30 to 40 percent of the men and 27 to 34 percent of the women who perpetrated violence against their partners were drinking at the time of the event. Alcohol-related problems were associated with IPV among blacks and whites, but not among Hispanics. Alcohol's role in partner violence may be explained by people's expectations that alcohol will have a disinhibitory effect on behavior or by alcohol's direct physiological disinhibitory effect. It is also possible that people consciously use alcohol as an excuse for their violent behavior or that alcohol appears to be associated with violence because both heavier drinking and violence have common predictors, such as an impulsive personality. PMID- 11496969 TI - Alcohol and violence in the lives of gang members. AB - Life within a gang includes two endemic features: violence and alcohol. Yet, to date, most researchers studying gang behavior have focused on violence and its relationship to illicit drugs, largely neglecting the importance of alcohol in gang life. Because alcohol is an integral and regular part of socializing within gang life, drinking works as a social lubricant, or social glue, to maintain not only the cohesion and social solidarity of the gang, but also to affirm masculinity and male togetherness. In addition to its role as a cohesive mechanism, particular drinking styles within gangs may operate, as with other social groups, as a mechanism to maintain group boundaries, thereby demarcating one gang from another. Other examples of internal gang violent activities associated with drinking include fighting between members because of rivalries, tensions, or notions of honor or respect. At a more symbolic level, drinking is associated with two important ritual events in gang life: initiation, or "jumping in,"and funerals. By better understanding the link between drinking and violence among youth gangs, steps can be taken to determine the social processes that occur in the development of violent behavior after drinking. PMID- 11496970 TI - Self-reported alcohol use and abuse by arrestees in the 1998 Arrestee Drug Abuse Monitoring Program. AB - Data collected in the Arrestee Drug Abuse Monitoring (ADAM) program on alcohol and other drug use among arrestees provide a valuable opportunity to examine the relationship between alcohol use and violence. The data are used to explore the combined use of alcohol and other drugs among offenders and the relationships between substance use and the offenders' demographic characteristics and offenses. These findings are used to identify changes in the offenders' alcohol and other drug use over time. PMID- 11496971 TI - The potential of inorganic mass spectrometry in mineral and trace element nutrition research. AB - Over the past two decades, new applications of inorganic mass spectrometry have been made possible by the use of stable isotopes as tracers in studies of mineral and trace element metabolism in man. Stable isotope techniques and radioisotope methods are the only reliable tools available for determination of the absorption, retention, or utilization of a nutrient by the human body. Recent developments in inorganic mass spectrometry might open new perspectives as progress in this field of research depends mainly on improving existing stable isotope techniques and on developing novel concepts. By improving precision in isotope analysis, isotope doses in experiments on man can be reduced to physiologically more meaningful levels. This will also enable reduction of the (often substantial) costs of isotopically labeling a nutrient in a test meal. Improvements in the mass spectrometric sensitivity will enable the development of new tracer techniques that have the potential to provide the information required by: 1. governmental institutions for designing food fortification programs; 2. the food industry for developing nutrient-fortified food products; and 3. public health authorities for establishing reliable dietary recommendations for intake of inorganic nutrients. In this context the current scope and limitations of thermal ionization mass spectrometry, inductively coupled mass spectrometry, accelerator mass spectrometry, and resonance ionization mass spectrometry are evaluated. Iron isotopic variations in the human body are discussed as a possible source of bias that might be a future biological limit to stable isotope-dose reduction in experiments on iron metabolism in man. PMID- 11496972 TI - Reaction chemistry and collisional processes in multiple devices for resolving isobaric interferences in ICP-MS. AB - A low-level review of the fundamentals of ion-molecule interactions is presented. These interactions are used to predict the efficiencies of collisional fragmentation, energy damping and reaction for a variety of neutral gases as a function of pressure in a rf-driven collision/reaction cell. It is shown that the number of collisions increases dramatically when the ion energies are reduced to near-thermal (< 0.1 eV), because of the ion-induced dipole and ion-dipole interaction. These considerations suggest that chemical reaction can be orders of magnitude more efficient at improving the analyte signal/background ratio than can collisional fragmentation. Considerations that lead to an appropriate selection of type of gas, operating pressure, and ion energies for efficient operation of the cell for the alleviation of spectral interferences are discussed. High efficiency (large differences between reaction efficiencies of the analyte and interference ions, and concomitant suppression of secondary chemistry) might be required to optimize the chemical resolution (determination of an analyte in the presence of an isobaric interference) when using ion molecule chemistry to suppress the interfering ion. In many instances atom transfer to the analyte, which shifts the analytical m/z by the mass of the atom transferred, provides high chemical resolution, even when the efficiency of reaction is relatively low. Examples are given of oxidation, hydroxylation, and chlorination of analyte ions (V+, Fe+, As+, Se+, Sr+, Y+, and Zr+) to improve the capability of determination of complex samples. Preliminary results are given showing O-atom abstraction by CO from CaO+ to enable the determination of Fe in high-Ca samples. PMID- 11496973 TI - Potential of mass spectrometry for the analysis of inorganic high-temperature vapors. AB - Mass spectrometry is the most versatile method for the analysis of high temperature vapors, because it enables the identification of the gaseous species and the determination of their partial pressures. If the vaporization processes are conducted under thermodynamic equilibrium, thermodynamic data can be evaluated from the partial pressures and their temperature-dependencies. The mass spectrometric methods used for the determination of thermodynamic data of high temperature vapors are Knudsen effusion mass spectrometry (KEMS), transpiration mass spectrometry (TMS), and laser-induced vaporization mass spectrometry (LVMS). KEMS is used whenever possible. Limitations of KEMS and TMS are the container problem, which limits the measurement temperatures to approximately 2500 K. The container problem is overcome by LVMS, which enables measurements up to approximately 7,000 K. The upper limit of the partial pressure measurement by KEMS of approximately 10 Pa does not apply for TMS, which enables measurements up to pressures of approximately 0.1 MPa. The fundamentals of the different methods are described and results are presented. Emphasis is on KEMS. PMID- 11496974 TI - Thorium and iodine memory effects in inductively-coupled plasma mass spectrometry. AB - Thorium and iodine memory effects have been characterized experimentally for inductively-coupled plasma mass spectrometry by adding ammonia gas directly to the spray chamber and nebulizing aqueous ammonia sample solutions to assess analyte memory retention sites. Thorium memory effect originates from the tendency of an unidentified thorium compound to volatilize from the spray-chamber walls, and not from Th compound adsorption to nebulizer tubing. The mass spectrometer skimmer and sampler cones, ion optics, quadrupole, and other components are not responsible for the memory effect. Unlike that of thorium the iodine memory effect originates from adsorption of iodine compounds on nebulizer tubing surfaces and from volatilization of HI and I2 from the spray-chamber walls. Addition of ammonia sample solutions or ammonia gas directly to the spray chamber eliminated the Th and I memory effects in practical analyses, and blank levels were achieved after 2 min wash-outs. Quantitative recoveries were obtained for Th and I in reference materials. PMID- 11496975 TI - Experimental evidence for the formation of doubly charged oxide and hydroxide ions in inductively coupled plasma mass spectrometry. AB - The formation of doubly charged polyatomic ions in inductively coupled plasma mass spectrometers was investigated using commercially available instruments. The species observed were ThO2+ and ThOH2+, which were found in similar amounts with the different instruments used in this study, when operated under routine analytical conditions. The signal ratios for ThO2+ were between 1.8 x 10(-4) and 4.2 x 10(-4) relative to the singly charged elemental ion and between 1.4 x 10( 2) and 2.2 x 10(-2) relative to the doubly charged elemental ion. The formation of ThOH2+, was between 1.1 x 10(-4) and 2.8 x 10(-4) relative to the singly charged elemental ion and between 0.72 x 10(-2) and 1.3 x 10(-2) relative to the doubly charged elemental ion. A mechanism is proposed for the formation of the doubly charged oxide and hydroxide ions that is based on the condensation of the doubly charged elemental ion with water or oxygen molecules in the interface region of the mass spectrometer. PMID- 11496976 TI - Removal of interfering elements in ICP-QMS for the determination of Pt, Rh, and Pd by chemically modified sample introduction capillaries. AB - New on-line methods developed for the determination of Pt, Rh, and Pd by inductively coupled plasma-quadrupole mass spectrometry (ICP-QMS) include separation of elements which might lead to spectral interference in the quadrupole instrument. The fused-silica capillaries generally used for transport of the sample to mu-flow nebulizers have been chemically modified with ion exchanger compounds to remove interfering elements such as Cu, Pb, or Hf. Characterization of the modification procedures by atomic-force microscopy showed that the quality of the quartz material and the kind of modification had a decisive influence on the yield of surface modification, and thus the exchange capacity of the capillaries. PMID- 11496977 TI - SI-traceable certification of the amount content of cadium below the ng g(-1) level in blood samples by isotope dilution ICP-MS applied as a primary method of measurement. AB - The development and implementation of a method for the certification of cadmium in blood samples at low ng g(-1) and sub ng g(-1) levels is described. The analytical procedure is based on inductively coupled plasma isotope dilution mass spectrometry (ICP-IDMS) applied as a primary method of measurement. Two different sample digestion methods, an optimized microwave digestion procedure using HNO3 and H2O2 as oxidizing agents and a high-pressure asher digestion procedure, were developed and compared. The very high salt content of the digests and the high molybdenum content, which can cause oxide-based interferences with the Cd isotopes, were reduced by a chromatographic matrix separation step using an anion exchange resin. All isotope ratio measurements were performed by a quadrupole ICP MS equipped with an ultrasonic nebulizer with membrane desolvator. This sample introduction set-up was used to increase sensitivity and minimize the formation of oxides (less MoO+ interference with the Cd isotopes). Because of the very low Cd concentrations in the samples and the resulting need to minimize the procedural blank as much as possible, all sample-processing steps were performed in a clean room environment. Detection limits of 0.005 ng g(-1) Cd were achieved using sample weights of 2.7 g. The method described was used to recertify the cadmium content of three different blood reference materials from the Community Bureau of Reference (BCR) of the European Commission (BCR-194, BCR-195, BCR- 196). Cadmium concentrations ranged between approximately 0.2 ng g(-1) and approximately 12 ng g(-1). For these materials, SI-traceable certified values including total uncertainty budgets according to ISO and Eurachem guidelines were established. PMID- 11496978 TI - Development of an electrothermal vaporization ICP-MS method and assessment of its applicability to studies of the homogeneity of reference materials. AB - A method has been developed for measurement of the homogeneity of analyte distribution in powdered materials by use of electrothermal vaporization with inductively coupled plasma mass spectrometric (ETV-ICP-MS) detection. The method enabled the simultaneous determination of As, Cd, Cu, Fe, Mn, Pb, and Zn in milligram amounts of samples of biological origin. The optimized conditions comprised a high plasma power of 1,500 W, reduced aerosol transport flow, and heating ramps below 300 degrees C s(-1). A temperature ramp to 550 degrees C ensured effective pyrolysis of approximately 70% of the organic compounds without losses of analyte. An additional hold stage at 700 degrees C led to separation of most of the analyte signals from the evaporation of carbonaceous matrix compounds. The effect of time resolution of signal acquisition on the precision of the ETV measurements was investigated. An increase in the number of masses monitored up to 20 is possible with not more than 1% additional relative standard deviation of results caused by limited temporal resolution of the transient signals. Recording of signals from the nebulization of aqueous standards in each sample run enabled correction for drift of the sensitivity of the ETV-ICP-MS instrument. The applicability of the developed method to homogeneity studies was assessed by use of four certified reference materials. According to the best repeatability observed in these sample runs, the maximum contribution of the method to the standard deviation is approximately 5% to 6% for all the elements investigated. PMID- 11496979 TI - 41Ca ultratrace determination with isotopic selectivity > 10(12) by diode-laser based RIMS. AB - 41Ca ultratrace determination by diode-laser-based resonance ionization mass spectrometry with extremely high isotopic selectivity is presented. Application to environmental dosimetry of nuclear reactor components, to cosmochemical investigations of production cross sections, and biomedical isotope-tracer studies of human calcium kinetics are discussed. Future investigations are possible use in 41Ca-radiodating. Depending on the application, 41Ca isotopic abundances in the range of 10(-9) to 10(-15) relative to the dominant stable isotope 40Ca must be determined. Either double- or triple-resonance optical excitation with narrow-band extended cavity diode lasers and subsequent non resonant photoionization of calcium in a collimated atomic beam were used. The resulting photoions are detected with a quadrupole mass spectrometer optimized for background reduction and neighboring mass suppression. Applying the full triple-resonance scheme provides a selectivity of approximately 5 x 10(12) in the suppression of neighboring isotopes and > 10(8) for isobars, together with an overall detection efficiency of approximately 5 x 10(-5). Measurements on a variety of sample types are discussed; the accuracy and reproducibility of the resulting 41Ca/40Ca isotope ratios was better than 5%. PMID- 11496980 TI - Investigations on the use of chemical modifiers for the direct determination of trace impurities in Al2O3 ceramic powders by slurry electrothermal evaporation coupled with inductively-coupled plasma mass spectrometry (ETV-ICP-MS). AB - The direct determination of trace impurities in Al2O3 ceramic basic powders by ICP-MS using electrothermal evaporation (ETV) with slurry sampling has been investigated. To increase interference-free analyte volatilization, the use of the palladium-group modifiers (PGM) IrCl3, Pd(NO3)2, and PdCl2 for the determination of Ca, Fe, Ga, Mg, Mn, Na, Ni, and V in Al2O3 powders was studied. Their role, which in ETV-ICP-MS and ETV-ICP-OES is to stabilize the investigated analyte during the ashing phase, to increase vaporization of the matrix, and to reduce transport losses was investigated. Optimum analysis results were obtained with PdCl2 modifier when 500 ng Pd was used for a sample weight of 100 microg Al2O3 injected into the ETV. Calibration was performed by standard addition with aqueous solutions of the analytes. The RSDs calculated from triplicate analysis ranged form 5 to 10%. Detection limits between 0.07 microg g(-1) (Ga) and 1.1 microg g(-1) (Na) were achieved. The accuracy was proven for the elements Ca, Fe, Ga, Mg, Mn, Na, Ni, and V by analyzing an NIST standard reference Al2O3 material (SRM 699) with a middle grain size of 16.4 microm. The analytical method was used for the analysis of Al2O3 powder (AKP 30, Sumitomo, Japan) with impurities in the low microg g(-1) range and a middle grain size of 1.1 microm. The results obtained for the elements Ca, Fe, Ga, Mg, Mn, Na, Ni, and V were comparable with those obtained by ICP-MS subsequent to conventional decomposition with hydrochloric acid at high pressure. PMID- 11496981 TI - Determination of trace elements in quartz glass by use of LINA-Spark-ICP-MS as a new method for bulk analysis of solid samples. AB - The determination of trace elements in pure quartz glass samples has been performed by coupling an ICP quadrupole mass spectrometer with the LINA-Spark Atomizer, an IR laser ablation system dedicated to direct bulk and surface analysis of solid samples. Linear calibration curves were obtained for nine elements (Na, Al, Ca, Ti, Cr, Mn, Zr, Ba, and Pb) in the ng g(-1) range with detection limits of less than 10 ng g(-1) for Ca, Cr, Mn, Zr, Ba, and Pb and in the range of 120-220 ng g(-1) for Na, Al, and Ti. The distance between the laser focal point and the sample surface has a significant influence on signal intensity and precision, both of which can be improved by a factor of approximately two by focusing the laser 15 mm behind the sample surface. Aerosol moistening reduced the standard deviation of the signal intensity by a factor of 2-4. Signal instability, which resulted from different ablation rates or variations in the transmission of the mass spectrometer, were compensated by use of the simultaneously measured SiAr+ ion as an internal standard. Under these conditions precision was usually better than 5% RSD. The results were compared with those obtained by use of a commercial LA-ICP-MS system. With this instrumentation linear calibration curves were achieved for three elements only (Al, Ti, and Pb), showing that LA-ICP-MS is less appropriate for bulk analysis in the ng g(-1) range. PMID- 11496982 TI - Determination of stoichiometry and concentration of trace elements in thin BaxSr1 xTiO3 perovskite layers. AB - This paper describes an analytical procedure for determining the stoichiometry of BaxSr1-xTiO3 perovskite layers using inductively coupled plasma mass spectrometry (ICP-MS). The analytical results of mass spectrometry measurements are compared to those of X-ray fluorescence analysis (XRF). The performance and the limits of solid-state mass spectrometry analytical methods for the surface analysis of thin BaxSr1-xTiO3 perovskite layers sputtered neutral mass spectrometry (SNMS)--are investigated and discussed. PMID- 11496983 TI - Trace analysis of high-purity graphite by LA-ICP-MS. AB - Laser-ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has been established as a very efficient and sensitive technique for the direct analysis of solids. In this work the capability of LA-ICP-MS was investigated for determination of trace elements in high-purity graphite. Synthetic laboratory standards with a graphite matrix were prepared for the purpose of quantifying the analytical results. Doped trace elements, concentration 0.5 microg g(-1), in a laboratory standard were determined with an accuracy of 1% to +/- 7% and a relative standard deviation (RSD) of 2-13%. Solution-based calibration was also used for quantitative analysis of high-purity graphite. It was found that such calibration led to analytical results for trace-element determination in graphite with accuracy similar to that obtained by use of synthetic laboratory standards for quantification of analytical results. Results from quantitative determination of trace impurities in a real reactor-graphite sample, using both quantification approaches, were in good agreement. Detection limits for all elements of interest were determined in the low ng g(-1) concentration range. Improvement of detection limits by a factor of 10 was achieved for analyses of high-purity graphite with LA-ICP-MS under wet plasma conditions, because the lower background signal and increased element sensitivity. PMID- 11496984 TI - Spark-source mass spectrometric assessment of silicon concentrations in silicon doped gallium arsenide single crystals. AB - The spark-source mass spectrometric assessment of silicon concentrations in silicon-doped vertical-gradient-freeze gallium arsenide is presented. The silicon concentrations determined are compared with the charge-carrier densities measured by means of the Hall effect with van der Pauw symmetry along the axis of a single crystal. PMID- 11496985 TI - Determination of rare earth elements in environmental matrices by sector-field inductively coupled plasma mass spectrometry. AB - In the framework of an international certification campaign, sector-field inductively coupled plasma mass spectrometry (sector-field ICP-MS) was used for the accurate determination of the rare earth elements in five candidate reference materials: aquatic plant, calcareous soil, mussel tissue, river sediment, and tuna muscle. All samples were taken into solution by use of microwave-assisted or mixed microwave-assisted / open beaker acid digestion. Subsequently, the samples were appropriately diluted and subjected to ICP-MS analysis. Except for Sc, all the elements involved were determined at low mass resolution (R = 300). For Sc, application of a higher resolution setting (R = 3,000) was required to separate the analyte signal from those of several molecular ions which gave rise to spectral overlap at low mass resolution. Some of the heavier REE can also suffer from spectral overlap attributed to the occurrence of oxide ions (MO+) of the lighter REE and Ba. This spectral overlap could be successfully overcome by mathematical correction. Matrix effects were overcome by use of two carefully selected internal standards, such that external calibration could be used. On each occasion, a geological reference material was analyzed as a quality-control sample and the reliability of all results obtained was additionally checked by means of chondrite normalization. For tuna muscle the content of all REE was below the limit of detection. For calcareous soil and river sediment, low to sub microg g(-1) values were observed, whereas the REE content of aquatic plant and mussel tissue was considerably lower (low to sub ng g(-1)). Overall, the results obtained were in excellent agreement with the average values, calculated on the basis of all "accepted" values, obtained in different laboratories using different techniques. PMID- 11496986 TI - Development of a procedure for the multi-element determination of trace elements in wine by ICP-MS. AB - An inductively coupled plasma mass spectrometric (ICP-MS) procedure has been developed for the determination of trace elements in wine. The procedure consists in simple 1+1 dilution of the wine and semi-quantitative analysis (without external calibration) using In as internal standard. Thirty-one elements at concentrations ranging from 0.1 mg mL(-1) to 0.5 ng mL(-1) can be determined by ICP-MS analysis with and without digestion. It was investigated whether a matrix effect observed for EtOH in the wine matrix can be overcome by application of a micro-concentric nebulizer with a membrane desolvator (MCN 6000). The results obtained for the MCN 6000 are compared with those obtained by use of a conventional Meinhard nebulizer. It is shown that the observed matrix effect can only be compensated by use of an internal standard for the Meinhard nebulizer, but not for the MCN 6000. Results for ICP-MS are compared with those obtained by total reflection X-ray fluorescence spectrometry (TXRF). PMID- 11496987 TI - Analysis of Ir in Kofelsit rocks by inductively coupled plasma-sector-field mass spectrometry (ICP-SFMS). AB - Three different analytical strategies have been evaluated for the quantification of Ir in geological samples. Glassy rock samples from Kofels and reference material WGB-1 were analyzed directly by inductively coupled plasma sector field mass spectrometry (ICP-SFMS) at mass resolution 400 using membrane desolvation and at mass resolution 9500 without membrane desolvation. Matrix separation by anion-exchange pre-concentration was also investigated. The ultrasonic nebulizer USN6000AT+ (Cetac Technologies, Omaha, NE, USA) incorporating a membrane desolvation unit was used as the sample-introduction system. Sample preparation involved complete microwave-assisted acid digestion of the silicate matrix with HNO3-HCl-HF. The results obtained by the three methods of quantification were in good agreement, showing that oxide-type interferences were effectively eliminated solely by membrane desolvation. The limits of detection were 6 pg g for low resolution measurement with use of the membrane, 15 pg g(-1) at a mass resolution of 9500, and 59 pg g(-1) for the ion-exchange procedure. The ultimate precision obtained for the Kofelsit Ir data was, however, compromised by the small sample intake (0.3 g), because of the inhomogeneous distribution of Ir in geological samples. PMID- 11496988 TI - Recognizing heterogeneous distribution of platinum group elements (PGE) in geological materials by means of the Re-Os isotope system. AB - The identification of uncertainties caused by sample inhomogeneity, as distinct from those caused by sample preparation and measurement, is a challenging task. Use of chemometric methods to separate and estimate these contributions to the combined standard uncertainty of a measurement (uc) of an analytical result requires complex experiments. The difficulty of platinum group element measurement makes this task even more complex. But unless it can be demonstrated that sample inhomogeneity is the major contributor to the high variability of an analytical result one should be careful not to mistakenly attribute this to a nugget effect. In this contribution we are able to demonstrate in two special cases that irreproducible results (up to 90% RSD) for analysis of Os and Re in the pg g(-1) to ng g(-1) range are truly caused by a nugget effect and not by inadequacies of the analytical method. PMID- 11496989 TI - Precise isotope-ratio measurements of lead species by capillary gas chromatography hyphenated to hexapole Multicollector ICP-MS. AB - The precision and accuracy of lead isotope-ratio determination on a short transient signal has been assessed by coupling capillary gas chromatography to the Isoprobe (Micromass, UK), a single-focusing inductively coupled plasma mass spectrometer with multicollector detection. A T-piece connecting the GC transfer line to the torch enabled continuous aspiration of thallium solution for mass bias correction. The volatile lead species PbEt4 was derivatized from NIST isotopic certified lead standard SRM 981 and different amounts of PbEt4 dissolved in iso-octane were injected into the GC. Chromatograms were recorded in multicollection mode by use of Faraday cups; seven isotopes (204Pb, 206Pb, 207Pb, 208Pb, PMID- 11496990 TI - HPIC-UV-ICP-SFMS study of the interaction of cisplatin with guanosine monophosphate. AB - Interaction of cis-[Pt(NH3)2Cl2] (cisplatin) with 5'-guanosine monophosphate (5' GMP) has been investigated for the first time by on-line coupling of high performance ion chromatography (HPIC) to inductively coupled plasma sector field mass spectrometry (ICP-SFMS). The time-dependent reaction course of the cisplatin 5'-GMP system was followed after incubation under simulated physiological conditions by monitoring the decrease in the concentration of 5'-GMP and the increase in the concentration of formed adducts, on the basis of speciation analysis. Because of the two-step mechanism an intermediate mono adduct was observed together with the major product, the bis adduct cis-[Pt(NH3)2(GMP)2]2-. The data obtained correlated well with those from earlier studies employing orthogonal techniques such as capillary electrophoresis (CE). Furthermore, HPIC ICP-SFMS provided unambiguous stoichiometric information about the major GMP adduct. For this purpose the platinum-to-phosphorus ratio was determined by simultaneously measuring 31P and 195Pt. To separate significant interferences from 15N16O+, 14N16O1H+, 12C18O1H+, and 13C17O1H+ on 31P, high-mass resolution (m/deltam = 4,500) proved to be mandatory. The P/Pt signal ratio of 2/1 obtained corresponds to the molar ratio in the bis adduct cis-[Pt(NH3)2(GMP)2]2-. PMID- 11496991 TI - Species-specific isotope-ratio measurements of volatile tin and antimony compounds using capillary GC-ICP-time-of-flight MS. AB - The analytical performance of an axial inductively-coupled-plasma time-of-flight mass spectrometer (ICP-TOFMS) as a detector for fast transient chromatographic signals resulting from the coupling to capillary gas chromatography (CGC) was investigated. A cryotrapping GC-ICP-TOFMS method for the determination of volatile metal(loid) compounds (VOMs) in gases was used and the suitability of the TOF mass analyzer for multielemental speciation analysis and multi-isotope ratio determinations was studied in terms of accuracy and precision. Isotope ratios 118Sn/120Sn and 121Sb/123Sb have been determined in in-house gas standard atmospheres in Tedlar bags at two different levels (100 pg and 1 ng) for different elemental species (SnH4, MeSnH3, Me2SnH2, Me3SnH, BuSnH3, SbH3, and MeSbH2). A limitation arising from counting statistics in both detection modes could be shown. A solution containing rhodium (10 ng mL(-1)) and cadmium (40 ng mL(-1)) was introduced simultaneously to the GC outlet. Rhodium acts as a continuous internal standard and Cd is used for mass-bias correction (by measuring the 111Cd/113Cd ratio). The detection system in both pulse counting and analog mode was examined. The best attainable precision was established for Me2SnH2 (analog mode, 12 replicates, 1 ng, RSD 0.34%, accuracy 0.31%) whereas most other species ranged between 0.4 and 0.5% RSD if higher concentrations were used. The limitations of the pulse counting system are clearly seen, with peak heights of more than 2000 counts reaching saturation (for an integration time of 100 ms), which reduces the accuracy of isotope ratio determinations. A dozen VOM could be detected in an aged landfill gas sample; several unidentified Sn compounds were present. Although their isotope ratios are within the confidence value of the standards, it is not yet clear if the acquired precision is good enough to identify isotopic fractionation of metal(loid)s through biovolatilization processes. With the precision achieved, the combination of cryotrapping GC and ICP-TOFMS is a powerful tool for monitoring volatile multi element species in multi-tracer experiments and isotope dilution methodology. PMID- 11496992 TI - Gas chromatography--inductively coupled plasma--time-of-flight mass spectrometry for the speciation analysis of organolead compounds in environmental water samples. AB - The application of inductively coupled plasma--time-of-flight mass spectrometry for the speciation analysis of organolead compounds in environmental waters is described. Construction of the transfer line was achieved by means of a relatively simple and rapid coupling procedure. Derivatization of the ionic lead species was achieved by in-situ propylation with sodium tetrapropylborate; simultaneous extraction of the derivatized compounds in hexane was followed by separation and detection by capillary gas chromatography hyphenated to inductively coupled plasma-time-of-flight mass spectrometry. Detection limits for the different organolead species ranged from 10 to 15 fg (as Pb), corresponding to procedural detection limits between 50 and 75 ng L(-1), on the basis of a 50 mL snow sample, extraction with 200 microL hexane, and subsequent injection of 1 microL of the organic extract on to the column. The accuracy of the system was confirmed by additional analysis of the water samples by capillary gas chromatography coupled with microwave-induced plasma-atomic-emission spectrometry and the analysis of a standard reference material CRM 605 (road dust) with a certified content of trimethyllead. PMID- 11496993 TI - Characterization of neutron transmuted zinc traces in pure copper materials by isotope dilution mass spectrometry. AB - The neutron transmutation doping (NTD) of highly pure copper with zinc was investigated as a promising means of achieving controlled gradation of the zinc content in the range 1-20 microg g(-1). The doping process leads to the enrichment of two stable isotopes 64Zn and 66Zn in a ratio which differs from that of natural isotopic distribution. Mass spectrometric investigations by thermal ionization mass spectrometry (TIMS) were performed to validate the results obtained by gamma spectrometry. The investigations included both determination of the isotopic ratios of the doped zinc isotopes and the analysis of the accumulated zinc contents by isotope dilution (ID) analysis. Thereby a sample-specific correction of the blank could be performed because the isotope 68Zn was not influenced, because of the transmutation process. The results obtained by TIMS prove the strict proportionality of the doped zinc content, in the range 5 to 20 microg g(-1), to the neutron fluence. Comparison with gamma spectrometric results showed a very good agreement within the uncertainties. PMID- 11496994 TI - Determination of uranium isotopic composition and 236U content of soil samples and hot particles using inductively coupled plasma mass spectrometry. AB - As a result of the accident at the Chernobyl nuclear power plant (NPP) the environment was contaminated with spent nuclear fuel. The 236U isotope was used in this study to monitor the spent uranium from nuclear fallout in soil samples collected in the vicinity of the Chernobyl NPP. Nuclear track radiography was applied for the identification and extraction of hot radioactive particles from soil samples. A rapid and sensitive analytical procedure was developed for uranium isotopic ratio measurement in environmental samples based on double focusing inductively coupled plasma mass spectrometry (DF-ICP-MS) with a MicroMist nebulizer and a direct injection high-efficiency nebulizer (DIHEN). The performance of the DF-ICP-MS with a quartz DIHEN and plasma shielded torch was studied. Overall detection efficiencies of 4 x 10(-4) and 10(-3) counts per atom were achieved for 238U in DF-ICP-QMS with the MicroMist nebulizer and DIHEN, respectively. The rate of formation of uranium hydride ions UH+/U+ was 1.2 x 10( 4) and 1.4 x 10(-4), respectively. The precision of short-term measurements of uranium isotopic ratios (n = 5) in 1 microg L(-1) NBS U-020 standard solution was 0.11% (238U/235U) and 1.4% (236U/238U) using a MicroMist nebulizer and 0.25% (235U/238U) and 1.9% (236U/P38U) using a DIHEN. The isotopic composition of all investigated Chernobyl soil samples differed from those of natural uranium; i.e. in these samples the 236U/238U ratio ranged from 10(-5) to 10(-3). Results obtained with ICP-MS, alpha- and gamma-spectrometry showed differences in the migration properties of spent uranium, plutonium, and americium. The isotopic ratio of uranium was also measured in hot particles extracted from soil samples. PMID- 11496995 TI - ICP-MS with hexapole collision cell for isotope ratio measurements of Ca, Fe, and Se. AB - To avoid mass interferences on analyte ions caused by argon ions and argon molecular ions via reactions with collision gases, an rf hexapole filled with helium and hydrogen has been used in inductively coupled plasma mass spectrometry (ICP-MS), and its performance has been studied. Up to tenfold improvement in sensitivity was observed for heavy elements (m > 100 u), because of better ion transmission through the hexapole ion guide. A reduction of argon ions Ar+ and the molecular ions of argon ArX+ (X = O, Ar) by up to three orders of magnitude was achieved in a hexapole collision cell of an ICP-MS ("Platform ICP", Micromass, Manchester, UK) as a result of gas-phase reactions with hydrogen when the hexapole bias (HB) was set to 0 V; at an HB of 1.6 V argon, and argon-based ions of masses 40 u, 56 u, and 80 u, were reduced by approximately four, two, and five orders of magnitude, respectively. The signal-to-noise ratio 80Se/ 40Ar2+ was improved by more than five orders of magnitude under optimized experimental conditions. Dependence of mass discrimination on collision-cell properties was studied in the mass range 10 u (boron) to 238 u (uranium). Isotopic analysis of the elements affected by mass-spectrometric interference, Ca, Fe, and Se, was performed using a Meinhard nebulizer and an ultrasonic nebulizer (USN). The measured isotope ratios were comparable with tabulated values from IUPAC. Precision of 0.26%, 0.19%, and 0.12%, respectively, and accuracy of 0.13% 0.25%, and 0.92%, respectively, was achieved for isotope ratios 44Ca/ 40Ca and 56Fe/57Fe in 10 microg L(-1) solution nebulized by means of a USN and for 78Se/80Se in 100 microg L(-1) solution nebulized by means of a Meinhard nebulizer. PMID- 11496996 TI - Isotope-ratio measurements of lead in NIST standard reference materials by multiple-collector inductively coupled plasma mass spectrometry. AB - The capability of a second-generation Nu Instruments multiple collector inductively coupled plasma mass spectrometer (MC-ICP-MS) has been evaluated for precise and accurate isotope-ratio determinations of lead. Essentially the mass spectrometer is a double-focusing instrument of Nier-Johnson analyzer geometry equipped with a newly designed variable-dispersion ion optical device, enabling the measured ion beams to be focused into a fixed array of Faraday collectors and an ion-counting assembly. NIST SRM Pb 981, 982, and 983 isotopic standards were used. Addition of thallium to the lead standards and subsequent simultaneous measurement of the thallium and lead isotopes enabled correction for mass discrimination, by use of the exponential correction law and 205Tl/203Tl = 2.3875. Six measurements of SRM Pb-982 furnished the results 206Pb/204Pb = 36.7326(68), 207Pb/204Pb = 17.1543(30), 208Pb/204Pb = 36.7249(69), 207Pb/206Pb = 0.46700(1), and 208Pb/206Pb = 0.99979(2); the NIST-certified values were 36.738(37), 17.159(25), 36.744(50), 0.46707(20), and 1.00016(36), respectively. Direct isotope lead analysis in silicates can be performed without any chemical separation. NIST SRM 610 glass was dissolved and introduced into the MC-ICP-MS by means of a micro concentric nebulizer. The ratios observed were in excellent agreement with previously reported data obtained by TIMS and laser ablation MC ICP-MS, despite the high Ca/Pb concentration ratio (200/1) and the presence of many other elements at levels comparable with that of lead. Approximately 0.2 microg lead are sufficient for isotope analysis with ratio uncertainties between 240 and 530 ppm. PMID- 11496997 TI - Laser ablation inductively coupled plasma mass spectrometry: a new tool for trace element analysis in ice cores. AB - A new method for the detection of trace elements in polar ice cores using laser ablation with subsequent inductively coupled plasma mass spectrometry analysis is described. To enable direct analysis of frozen ice samples a special laser ablation chamber was constructed. Direct analysis reduces the risk of contamination. The defined removal of material from the ice surface by means of a laser beam leads to higher spatial resolution (300-1000 microm) in comparison to investigations with molten ice samples. This is helpful for the detection of element signatures in annual layers of ice cores. The method was applied to the successful determination of traces for the elements Mg, Al, Fe, Zn, Cd, Pb, some rare-earth elements (REE) and minor constituents such as Ca and Na in ice cores. These selected elements serve as tracer elements for certain sources and their element signatures detected in polar ice cores can give hints to climate changes in the past. We report results from measurements of frozen ice samples, the achievable signal intensities, standard deviations and calibration graphs as well as the first signal progression of 205Pb in an 8,000-year-old ice core sample from Greenland. In addition, the first picture of a crater on an ice surface burnt by an IR laser made by cryogenic scanning electron microscopy is presented. PMID- 11496998 TI - Studies of LA-ICP-MS on quartz glasses at different wavelengths of a Nd:YAG laser. AB - The capability of LA-ICP-MS for determination of trace impurities in transparent quartz glasses was investigated. Due to low or completely lacking absorption of laser radiation, laser ablation inductively coupled plasma mass spectrometry (LA ICP-MS) proves difficult on transparent solids, and in particular the quantification of measurement results is problematic in these circumstances. Quartz glass reference materials of various compositions were studied by using a Nd:YAG laser system with focused laser radiation of wavelengths of 1064 nm, 532 nm and 266 nm, and an ICP-QMS (Elan 6000, Perkin Elmer). The influence of ICP and laser ablation conditions in the analysis of quartz glasses of different compositions was investigated, with the laser power density in the region of interaction between laser radiation and solid surface determining the ablation process. The trace element concentration was determined via calibration curves recorded with the aid of quartz glass reference materials. Under optimized measuring conditions the correlation coefficients of the calibration curves are in the range of 0.9-1. The relative sensitivity factors of the trace elements determined in the quartz glass matrix are 0.1-10 for most of the trace elements studied by LA-ICP-MS. The detection limits of the trace elements in quartz glass are in the low ng/g to pg/g range. PMID- 11496999 TI - A simple method of target preparation for the bulk analysis of powder samples by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). AB - A simple and rapid procedure using a glue technique has been developed for the preparation of stable targets from powder samples for bulk analysis by LA-ICP-MS. The procedure was evaluated for the analysis of trace elements in SiC, of rare earth elements in different types of silicate (rocks, sediments, and soils), and of Au and platinum-group elements in geological silicates. The test analysis was conducted using an IR laser in combination with a quadrupole mass spectrometer. The recommended preparation procedure offers the possibility of different types of calibration, for example application of certified reference samples in combination with prepared spiked samples on a base of a natural or synthetic matrix, or addition calibration. The resulting calibration functions are linear over a range of several decades. The trueness of the results was evaluated by use of certified reference samples. Analytical concentration ranges, detection limits, and the relative standard deviations are reported. PMID- 11497000 TI - Progress in multi-ion counting spark-source mass spectrometry (MIC-SSMS) for the analysis of geological samples. AB - Spark source mass spectrometry (SSMS) has experienced important and significant improvements in nearly all analytical features by the use of a multiple ion counting (MIC) system. Two procedures have recently been developed to further increase the analytical capabilities of MIC-SSMS in geochemistry. These are a mathematical correction of interferences, which is often necessary for the ultra trace element analysis of Nb, Ta, Zr, Hf and Y, and the development of an autospark system to hold the total ion beam constant. New analytical data for geological samples, especially international reference materials, are presented using the improved MIC-SSMS technique. The data set consists of high precision and low abundance data for Zr, Nb and Y in depleted reference materials. The MIC SSMS results are compared with those of conventional SSMS using photoplates for ion detection. The precision of the MIC-SSMS isotope ratio measurements (about 1%) is more than a factor of 3 better than that of conventional SSMS, as demonstrated by analyses of Hawaiian samples. Total uncertainties of MIC-SSMS concentration data including all sources of error are generally between 2 and 5% for concentrations higher than about 0.3 microg/g and about 10% for trace element abundances in the ng/g range. PMID- 11497001 TI - Modeling of the sputtering process of cubic silver halide microcrystals and its relevance in depth profiling by secondary-ion mass spectrometry (SIMS). AB - Secondary-ion mass spectrometry is frequently used for concentration-depth profiling of macroscopic samples, but it is certainly not a common analytical technique for the analysis of sub-micrometer-size particles. This is because of the additional ion-bombardment-induced artifacts which can occur when a three dimensional microvolume is sputtered, instead of a flat surface. This paper presents a model of how small cubic photographic Ag(Cl,Br) crystals are eroded under primary-ion bombardment, and the extent to which secondary ions generated at different faces are extracted. The latter is studied by means of the program SIMION, which simulates ion trajectories in complex electrical field systems. It is shown that up to 90% of the secondary ions originating from the side face of a cubic crystal are unable to reach the detector, in contrast with most secondary ions originating from the top face. The angular dependence of the sputtering yield and the elemental ratio of Br/Cl sputtered particles have been calculated by using the well-known computer code TRIM (transport of ions in matter) under some limiting assumptions (possible preferential sputtering is disregarded and a steady-state sputtering process is assumed). The validity of the theoretical model and the calculated results were checked with experimental data. On the basis of the depth profiles presented it is explained why it is still possible to measure an interface inside a cubic volume, even though a group of several hundred crystals is sputtered simultaneously, and even though the orientations of the distinct faces of the cubes relative to the angle of incidence of the primary ion beam are different. PMID- 11497002 TI - Design and development of a new program for data processing of mass spectra acquired by means of a high-resolution double-focusing glow-discharge mass spectrometer. AB - An new program has been developed and implemented for data analysis of mass spectra obtained by use of the VG9000 glow-discharge mass spectrometer. The program, designed to run in a Windows 9X environment includes several tools for import and export of data, cluster generators, etc. An automated technique for the interpretation of mass spectra is also built into the program; this enables faster and operator-independent interpretation. When major interferences or not well defined signals are involved, the automated technique might fail to find the correct result. Therefore, a manual, VG9000 software-like, bypass is at hand. A comparison of the different techniques and programs shows, in general, comparable results. An installable version of the software is available on the university FTP-server (ftp://PLASMA-FTP.uia.ac.be/ private/imsas/). PMID- 11497003 TI - Restoration of a fractured primary incisor. AB - Esthetic restoration on primary teeth has been a special challenge to pediatric dentists. Composite restorations are the most often used treatment for decay and fractures of primary teeth, however, there are other possible alternatives. We present a case in which we have used an acrylic crown to treat a fractured primary incisor in a 1.8 years old child. PMID- 11497004 TI - Focal dermal hypoplasia: management of complex dental features. AB - A 17 year-old female who presented for treatment of grossly carious lower first molar teeth had multiple features of Focal Dermal Hypoplasia (FDH). These included enamel pitting and hypoplasia, anomalies of shape, size and positioning of teeth, as well as soft tissue papillomas and telangiectasis of tongue. A case report and a synopsis of the syndrome (FDH) are presented, the oral aspects of the condition are reviewed and the management of the complex dental anomalies is discussed. PMID- 11497005 TI - Oligodontia: a case report. AB - Oligodontia is the agenesis of numerous teeth, and is commonly associated with specific syndromes and/or severe systemic abnormalities. In this paper a patient with 14 missing permanent teeth is presented clinically with her conservative and prosthetic treatments. PMID- 11497006 TI - Pseudo-Class III treatment with reverse traction: case report. AB - The pseudo-Class III can be defined as a functional reflex of an anterior positioning of the mandible, an acquired muscular position that simulates a mesiocclusion. The diagnosis and treatment plan of this condition must be based on a cephalometric evaluation that provides information about the relative contributions of the skeletal and dental components to the malocclusion. There is still great controversies about when is the best moment to start the Class III treatment. The purpose of this article is to describe a case report in which a Class III patient was successfully treated with reverse traction. PMID- 11497007 TI - The functional treatment of anterior-open bite: three case reports. AB - Functional treatment of three skeletal open-bite patients with Bionator, Frankel IV and Posterior bite-block appliances is presented. Pretreatment and post treatment records were evaluated. Correction of malocclusion by changing direction of the mandibular development, adaptive bony development of the condyle, vertical development in posterior region, an increase in ramus length, a decrease in the lower facial height were observed in this report. The results of functional treatment of three appliances had different effect mechanisms. PMID- 11497008 TI - Comparison of the behavior of children undergoing restorative dental treatment at the first visit versus the second visit in a private pediatric dental practice. AB - The purpose of this study was to determine if there was a difference in the behavior of child patients undergoing restorative dental treatment at the first office visit versus those whose first restorative treatment visit was after an initial non-threatening dental visit in a private pediatric dental practice. For patients up to and including age 9, patient behavior was recorded during the restorative session using the Sarnat scale, which rate behavior in 5 levels, from completely cooperative to completely uncooperative. Variables such as age, method of payment, referral source and sex were also recorded. The results showed that there was no statistically significant difference in the behavior of children, who had the first restorative dental experience at the initial office visit versus those children who had the first restorative procedure after a non invasive introductory visit in all instances. There were no differences according to age, sex, socio-economic status or source of referral. It is concluded that a child may not exhibit more negative behavior as a restorative dental patient when the first visit is for restorative therapy than if the restorative treatment is delivered at a later date after a non-threatening introduction to the dental environment. Thus, a pediatric dentist need not hesitate to treat a child at the first visit for fear that it may engender more negative behavior than if the restorative dental treatment was postponed until another time. PMID- 11497009 TI - Effects of carbamide peroxide containing bleaching agents on the morphology and subsurface hardness of enamel. AB - The purpose of this study is to evaluate the effects of nightguard bleaching agents (Karisma and Yotuel) on the enamel surface of forty anterior teeth. Ten teeth of each group were evaluated with SEM and ten teeth of each group were tested with a microhardness tester Morphologic alterations were observed on the enamel surfaces with SEM. Karisma group showed a significant decrease in enamel hardness (p<0.05) and microhardness values of enamel were increased significantly in Yotuel group (p<0.05). PMID- 11497010 TI - Wear of resin-modified glass ionomers: an in vitro study. AB - The purpose of this study was to evaluate the wear resistance and clinical applicability of resin-modified glass ionomer cements as restorative or fissure sealing materials. The in vitro wear of resin-modified glass ionomers was compared to conventional glass ionomers, a resin-based sealant, and a composite resin. A three-body wear test (enamel block--polymethylmethacrylate powder- experimental dental material) was performed by 20,000 cycles with a load of 4 kgf/cm2. The depth of wear of the experimental materials was measured and calculated using a computerized laser surface scanner. The glass ionomers generally showed more wear than the resin-based sealant and the composite resin, but there was no difference in wear between resin-modified and conventional glass ionomers. Type III ionomers (used for sealant) showed lower wear resistance than type II ionomers (used for restoration). PMID- 11497011 TI - Localization of first permanent molars in lateral cephalometric and panoramic radiographs from early mixed dentition to early permanent dentition. AB - Changes in position and mesiodistal angulations of maxillary and mandibular first permanent molars from early mixed dentition to early permanent dentition were measured on panoramic radiographs and compared with the values measured on lateral cephalometric radiographs. It was found that the maxillary and mandibular first permanent molars were uprighted gradually, drifted mesially and vertically, and that the changes in mesiodistal angulations were clearly on the panoramic radiographs in comparison with the cephalometric radiographs. PMID- 11497012 TI - pH changes in dental plaque after using sugar-free pediatric medicine. AB - The aim of this study was to compare the pH changes in the dental plaque after rinsing with sugared- (sucrose) or sugar-free (saccharine, cyclamate and sorbitol) versions of the same pediatric acetaminophen solution (ekosetol) up to one hour. Twenty-nine undergraduate dental students (17 girls and 12 boys) collected plaque for 48 hours by abstaining from oral hygiene, during which period they maintained normal dietary habits. Plaque sampling was done in two subsequent days before and 2, 5, 10, 15, 20, 25, 30, 40, 50 and 60 minutes after rinsing with both solutions. Measurements of pH were done within one hour using a micro pH electrode and a pH meter. All experiments were finished in 6 days by dividing the group to three. Results showed a significant difference between groups in respect to pH values and pH drops after rinsing. Mean pH values were below 5.70 for one hour in sugared solution, whereas no mean pH value was detected below 5.80 for one hour with the sugar-free solution. Minimum pH values (sugar-free: 5.62+/-0.36; sugared: 5.00+/-0.33, p<0.001) and maximum pH drops (sugar-free: -0.57+/-0.26; sugared: -1.16+/-0.44, p<0.001) were also significantly different. No difference was found between genders. We concluded that changing of sucrose to non-acidogenic sweeteners was essential to prevent the cariogenic potential of the pediatric medicines. PMID- 11497013 TI - Sotos syndrome with enamel hypoplasia: a case report. AB - A rare case of Sotos syndrome with enamel hypoplasia is described. Dental abnormalities include enamel hypoplasia, expansion of the pulp cavity, high arched palate, and absence of the bilateral pre-molar teeth of the mandible. PMID- 11497014 TI - Oral manifestations of Menkes' kinky hair syndrome. AB - Menkes' Kinky Hair Syndrome (MKHS) comprises an array of clinical manifestations including hair shaft abnormalities, epidermal hypopigmentation, and progressive cerebral degeneration that are transmitted as an X-linked recessive disorder affecting copper transport pathways in primarily young males. The oral manifestations of MKHS are scantly reported to include the presence of gingival enlargement and delayed eruption of primary teeth. The purpose of this report is to present a case of MKHS describing the intraoral clinical findings. PMID- 11497015 TI - Autistic children: experience and severity of dental caries between 1980 and 1995 in Kagoshima City, Japan. AB - The present study indicates that experience and severity of dental caries in 1995 showed a clear decrease from observations in 1980 in Kaogsima City, Japan. This decline may be attributed to several factors such as the qualitative and quantitative improvements in treatment of caries, more regular visits to dental clinics, improved daily oral hygiene, and changes in dietary pattern. PMID- 11497016 TI - The effect of bone marrow transplantation on systemic and oral health in Fanconi's aplastic anemia. AB - Fanconi's anemia (FA) is an autosomal-recessive disorder characterized by a progressive pancytopenia, diverse congenital abnormalities and increased predisposition to malignancy. Sixteen children with FA, aged between 4 to 16 were divided into two groups according to treatments. Nine children had bone marrow transplantation and seven children were treated with steroid and/or anapolan. The changes in dental caries, caries-associated microflora, salivary status and periodontal health were investigated in children with FA. Data were analyzed by one-way ANOVA. A statistically significant difference was found in hematological findings between children who have received bone marrow transplantation (BMT+) and the others, who have not received (BMT-). There was no significant difference in dental caries experience, salivary flow rate, buffering capacity, mutans streptococci and Lactobacilli levels between the study groups. A statistically significant difference was found in gingival index, plaque index, bleeding on probing, probing depth scores between the patients with FA in BMT(+) and BMT(-) groups (p<.05). In conclusion, besides systemic control, additional preventive measures during their whole life to maintain oral health is necessary in these children. PMID- 11497017 TI - Hyper-IgE syndrome: a case report. AB - The hyper-IgE syndrome (HIES) is a rare disorder characterized by pruritic dermatitis, recurrent Staphylococcus skin abscesses and extremely elevated levels of IgE in serum. In this report, an eleven-year-old-boy with hyper-IgE syndrome is presented. He had a coarse facial appearance, pruritic dermatitis, recurrent skin abscesses, pulmonary infection, a reduced rate of resorption of the roots of primary teeth and an elevated serum IgE concentration. The colonization of Candida albicans, Kiebsiella pneumoniae, Escherichia coli and Staphylococcus aureus were found as; (1x10(2) CFU), (2.2x10(4) CFU), (2.2x10(4) CFU) and (2.6x10(3) CFU) per ml saliva, respectively. Also the pulp of a deciduous molar was investigated with light and transmission electron microscope (TEM). As conclusion, treatment for this condition is lifelong administration of therapeutic doses of a penicillinase-resistant penicillin, with the addition of other antibiotics or anti-fungal agents as required for specific infections. PMID- 11497018 TI - Pediatric dental procedures under general anesthesia at the Amiri Hospital in Kuwait. AB - Ninety-six patients, fifty-eight healthy patients and thirty-eight special needs patients, ranging in age from 3 to 31 yr (average age 6.9 yr) had received dental treatments under general anesthesia (GA) at Amiri Hospital. The mean number of dental procedures per child was 13.6 (SD+/-5.4) with a mean number of 15.6 procedures for healthy patients and 10.5 special needs patients. There was a statistically significantly higher number of dental procedures for healthy patients than for special needs patients (p<0.001). The number of pulpotomies and stainless steel crowns placed for healthy patients was significantly higher (3.5) than for special needs patients (1.3) with (p<0.001) The average number of extractions was similar for the two groups of patients, 2.8 for normal and 2.2 for handicapped. On the basis of these results, it was concluded that dental treatment under GA in hospital environment is beneficial for certain group of patients, such as very young children and those with special needs. Underlying medical conditions influenced the treatment provided. PMID- 11497019 TI - Introduction: risk-adjustment issues in mental health services. AB - State mental health authorities and other public and private entities are developing outcome measures and comparing results across providers, programs, and systems. To make comparisons equitable, outcomes must be risk adjusted. This article provides an introduction to mental health risk adjustment and outlines issues involved in the selection of outcome and risk variables, data collection protocols, and analytic methods. It stresses the importance of proper identification of risk-adjustment variables and models. The article concludes with the next steps necessary to develop a valid approach to the risk-adjustment methodology. PMID- 11497020 TI - Approaches to risk-adjusting outcome measures applied to criminal justice involvement after community service. AB - The ethic of fairness in program evaluation requires that measures of behavioral health agency performance be sensitive to differences in those agencies' caseload composition. The authors describe two traditional approaches to the statistical risk adjustment of outcome measures (stratification weighting and pre-post measurement) that are designed to account for differences in caseload composition and introduce a method that incorporates the strengths of both approaches. Procedures for deriving each of these measures are described in detail and demonstrated in the evaluation of a statewide system of community-based behavioral health care programs. This evaluation examines the degree to which service recipients get into trouble with the law after treatment. Three measures are recommended for inclusion in outcome-oriented "report cards," and the interpretation of each measure is discussed. Finally, the authors suggest formats for graphic and tabular presentation of the risk-adjusted evaluation for sharing findings with diverse stakeholder groups. PMID- 11497021 TI - Comparing alternative risk-adjustment models. AB - The use of mental health indicators to compare provider performance requires that comparisons be fair. Fair provider comparisons mean that scores are risk adjusted for client characteristics that influence scores and that are beyond provider control. Data for the study are collected from 336 outpatients receiving publicly funded mental health services in Washington State. The study compares alternative specifications of multiple regression-based risk-adjustment models to argue that the particular form of the model will lead to different conclusions about comparative treatment agency performance. In order to evaluate performance fairly it is necessary to not only incorporate risk adjustment, but also identify the most correct form that the risk-adjustment model should take. Future research is needed to specify; test, and validate the mental health risk-adjustment models best suited to particular treatment populations and performance indicators. PMID- 11497022 TI - Risk adjustment of Florida mental health outcomes data: concepts, methods, and results. AB - This article discusses outcome evaluation systems for mental health programs. It reviews and critically evaluates design and analysis methods for strengthening the validity of such uncontrolled comparisons. The article examines methods for statistically adjusting preexisting groups, now referred to as risk adjustment or case-mix adjustment, and offers guidelines for determining when this procedure is appropriate. Then, analyses on two dependent variables--a global rating of functioning and a consumer satisfaction measure--available from an outcomes evaluation system currently underway in Florida are used to demonstrate the proposed method of risk adjustment. Results for 24 providers of mental health services showed that while risk adjustment only made a small difference in the overall provider rankings, the ranking of some specific providers changed considerably. The article concludes with a discussion of the implications of this research. PMID- 11497023 TI - Measuring mental health outcomes with pre-post designs. AB - The pre-post design has been the workhorse of outcome evaluations for many years. Using data from a study of 984 treated children (ages 5 to 17 years), this article argues that there are two structural problems with the pre-post evaluation of outcome: (1) excessively large intervals of uncertainty for individual outcomes and (2) paradoxical inconsistencies in the evaluation of groups. These problems can be solved by designs with three or more repeated measures analyzed with longitudinal multilevel analytic models. PMID- 11497024 TI - Comparing outcomes of routine care for depression: the dilemma of case-mix adjustment. AB - The purpose of this study was to formulate and test two case-mix models for depression treatment that permit comparisons of patient outcomes across diverse clinical settings. It assessed demographics; eight, diagnostic-specific, case-mix variables; and clinical status at baseline and follow-up for 187 patients. Regressions were performed to test two models for four dependent variables including depression severity and diagnosis. Individual treatment settings were then ranked based on a comparison of actual versus predicted outcomes using regression coefficients and predictor variables. A model inclusive of baseline physical health status and depression severity predicted depression severity, mental health, and physical health functioning at follow-up. A simpler model performed well in predicting depression remission. This study identifies variables to be included in case-mix adjustment models and demonstrates statistical methods to control for differences across settings when comparing depression outcomes. PMID- 11497025 TI - Risk adjustment in the Hoosier Assurance Plan: impact on providers. AB - The Indiana Division of Mental Health designed a method for developing risk adjusted case rates built on multidimensional assessment instruments that identified a consumer's level of need. An algorithm was developed sorting consumers within each of three target populations (adults with serious mental illness, adults with a chronic addiction, and children/adolescents with serious emotional disorders) into groups based on the relationship between their level of need and their similarity with regard to service cost. Actuarially determined risk-adjusted case rates were assigned to each group and applied to the case mix served by each provider in Indiana's public behavioral health system. This article describes four studies through which a determination was made as to the applicability of the risk-adjustment methodology. The results of the four studies supported using the adjusted case rates for adult consumers, but not for children/adolescents. PMID- 11497026 TI - National estimates of mental health utilization and expenditures for children in 1998. AB - No recent national data on expenditures and utilization are available to provide a benchmark for reform of mental health systems for children and adolescents. The most recent estimates, from 1986, predate the dramatic growth of managed care. This study provides updated national estimates. Treatment expenditures are estimated to be $11.68 billion ($172 per child). Adolescents have the highest expenditures at $293 per child followed by $163 per child aged 6 to 11 and $35 per preschool-aged child. Outpatient services account for 57%, inpatient for 33%, and psychotropic medications for 9% of the total. Unlike earlier reports, outpatient care now accounts for the majority of expenditures. This finding replicates the differences between recent managed care data and earlier actuarial databases for privately insured adults and confirms the trend from inpatient toward outpatient care. PMID- 11497027 TI - Insurance status and length of stay for involuntarily hospitalized patients. AB - General and private psychiatric hospitals are becoming increasingly common as sites for involuntary hospitalization. Unlike the public facilities that these settings are supplanting, these hospitals must pay strict attention to issues associated with reimbursement, insurance status, and managed care. This article examines the effects of insurance status on length of stay for involuntarily hospitalized patients in general and private hospitals in Massachusetts. Using a two-stage sampling procedure, data on episodes of involuntary hospitalization were gathered and assessed using multiple regression. The primary effect was found between patients with Medicare, who had the longest stays, and individuals who were uninsured, who had the shortest. The data raise concerns that warrant closer scrutiny on the part of administrators and clinicians. PMID- 11497029 TI - Sources of structure: genetic, environmental, and artifactual influences on the covariation of personality traits. AB - The phenotypic structure of personality traits has been well described, but it has not yet been explained causally. Behavior genetic covariance analyses can identify the underlying causes of phenotypic structure; previous behavior genetic research has suggested that the effects from both genetic and nonshared environmental influences mirror the phenotype. However, nonshared environmental effects are usually estimated as a residualterm that may also include systematic bias, such as that introduced by implicit personality theory. To reduce that bias, we supplemented data from Canadian and German twin studies with cross observer correlations on the Revised NEO Personality Inventory. The hypothesized five-factor structure was found in both the phenotypic and genetic/familial covariances. When the residual covariance was decomposed into true nonshared environmental influences and method bias, only the latter showed the five-factor structure. True nonshared environmental influences are not structured as genetic influences are, although there was some suggestion that they do affect two personality dimensions, Conscientiousness and Love. These data reaffirm the value of behavior genetic analyses for research on the underlying causes of personality traits. PMID- 11497030 TI - The non-evaluative circumplex of personality adjectives. AB - In judgments about personality, descriptive and evaluative aspects are ordinarily combined; separating them can be important both theoretically and practically. Study 1 showed that two similar descriptive factors can be found in analyses of personality terms, selected independently in English and in German and using different methods to control for evaluation. The factors relate to two pairs of independent axes suggested by previous work: Assertive-Unassertive and Tight Loose, or alternatively, Interactional Orientation (Extraversion-Introversion) and Affective Orientation. These two pairs of axes are shown to be rotations of each other, and to form the prime non-evaluative circumplex. As in previous studies, non-evaluative scales elicited higher levels of self-peer agreement than did more typical evaluation-confounded scales. Study 2 showed that adjective scales for the octants of this circumplex have circular ordering, can fit even very stringent constraints of a circumplex model, have mild to strong isomorphism with the interpersonal circumplex, but represent somewhat broader constructs, and are systematically related to the Big Five and the Big Three personality factors. PMID- 11497028 TI - Toward a national consumer survey: evaluation of the CABHS and MHSIP instruments. AB - This article describes a study evaluating the Consumer Assessment of Behavioral Health Survey (CABHS) and the Mental Health Statistics Improvement Program (MHSIP) surveys. The purpose of the study was to provide data that could be used to develop recommendations for an improved instrument. Subjects were 3,443 adults in six behavioral health plans. The surveys did not differ significantly in response rate or consumer burden. Both surveys reliably assessed access to treatment and aspects of appropriateness and quality. The CABHS survey reliably assessed features of the insurance plan; the MHSIP survey reliably assessed treatment outcome. Analyses of comparable items suggested which survey items had greater validity. Results are discussed in terms of consistency with earlier research using these and other consumer surveys. Implications and recommendations for survey development, quality improvement, and national policy initiatives to evaluate health plan performance are presented. PMID- 11497031 TI - The need for affect: individual differences in the motivation to approach or avoid emotions. AB - The present research developed and tested a new individual-difference measure of the need for affect, which is the motivation to approach or avoid emotion inducing situations. The first phase of the research developed the need for affect scale. The second phase revealed that the need for affect is related to a number of individual differences in cognitive processes (e.g., need for cognition, need for closure), emotional processes (e.g., affect intensity, repression-sensitization), behavioral inhibition and activation (e.g., sensation seeking), and aspects of personality (Big Five dimensions) in the expected directions, while not being redundant with them. The third phase of the research indicated that, compared to people low in the need for affect, people high in the need for affect are more likely to (a) possess extreme attitudes across a variety of issues, (b) choose to view emotional movies, and (c) become involved in an emotion-inducing event (the death of Princess Diana). Overall, the results indicate that the need for affect is an important construct in understanding emotion-related processes. PMID- 11497032 TI - A longitudinal study of personality change in young adulthood. AB - The present research examined personality continuity and change in a sample of young men and women assessed at the beginning and end of college. Two-hundred seventy students completed measures of the Big Five personality traits when they first entered college and then 4 years later. Analyses indicate small- to medium sized normative (i.e., mean-level) changes, large rank-order stability correlations, high levels of stability in personality structure, and moderate levels of ipsative (i.e. profile) stability. Overall, the findings are consistent with the perspective that personality traits exhibit considerable continuity over time, yet can change in systematic ways. PMID- 11497033 TI - Re-examining the general positivity model of subjective well-being: the discrepancy between specific and global domain satisfaction. AB - Three studies were conducted to examine the role of global life satisfaction in the discrepancy between specific and global domain satisfaction. Participants rated both global (e.g., education) and the corresponding, specific domain (e.g., professors, textbooks) satisfactions. In 3 studies, we found that individuals with higher life satisfaction evaluated global domain as a whole as more satisfying than those with lower life satisfaction, given the same level of satisfaction with specific domains. In Study 3, we also found that, given the same level of satisfaction during the previous 2 weeks, individuals with higher life satisfaction rated the global domains in general as more satisfying than those with lower life satisfaction. Overall, the association between globallife satisfaction and evaluative enhancement of global domains was most consistent in "self" and "social relationships." Finally, the effect of global life satisfaction on evaluative enhancement remained significant, controlling for extraversion and neuroticism. PMID- 11497034 TI - Eliminate nontherapeutic antimicrobial use in agriculture: AMA. PMID- 11497035 TI - The problem with pain. Veterinarians are making a thorny issue a priority. PMID- 11497036 TI - Despite concerns about job satisfaction and salaries, future looks bright for veterinary technicians. PMID- 11497037 TI - Washington State professor gets an education in Nepal. PMID- 11497038 TI - Thinks profession needs more holistic approach. PMID- 11497039 TI - Clinical observation: bilateral upward fixation of the patella following epidural administration of morphine. PMID- 11497040 TI - What are veterinarians worth? PMID- 11497041 TI - What is your diagnosis? Pronounced ventral deviation of the colon by an irregular soft tissue opacity in the retroperitoneal area. PMID- 11497042 TI - Promoting greater efficiency in the delivery of veterinary services. PMID- 11497043 TI - Semen sales and insemination--some legal issues. PMID- 11497044 TI - Epidemiologic and biological characteristics of salmonellosis in three dairy herds. PMID- 11497045 TI - Plasma concentration of ionized calcium in healthy iguanas. AB - OBJECTIVE: To measure plasma concentration of ionized calcium in healthy green iguanas. DESIGN: Prospective study. ANIMALS: 9 juvenile and 21 (10 male, 11 female) adult iguanas. PROCEDURE: Blood samples were obtained from each iguana, and plasma calcium, glucose, phosphorus, uric acid, total protein, albumin, globulin, potassium, and ionized calcium concentrations, aspartate transaminase (AST) activity, and pH were measured. Heparinized blood was used for measurement of ionized calcium concentration and blood pH. A CBC was also performed to assess the health of the iguanas. RESULTS: Significant differences were not detected among the 3 groups (juveniles, males, and females) with regard to ionized calcium concentration. Mean ionized calcium concentration measured in blood was 1.47 +/- 0.105 mmol/L. Significant differences were detected between juveniles and adults for values of phosphorus, glucose, total protein, albumin, globulin, and AST activity. CONCLUSIONS AND CLINICAL RELEVANCE: Ionized calcium concentration provides a clinical measurement of the physiologically active calcium in circulation. Evaluation of physiologically active calcium in animals with suspected calcium imbalance that have total plasma calcium concentrations within reference range or in gravid animals with considerably increased total plasma calcium concentrations is vital for determining a therapeutic plan. Accurate evaluation of calcium status will provide assistance in the diagnosis of renal disease and seizures and allow for better evaluation of the health status of gravid female iguanas. PMID- 11497046 TI - Nontraumatic rupture of an adrenal gland tumor causing intra-abdominal or retroperitoneal hemorrhage in four dogs. AB - Diagnosis and surgical management of intra-abdominal or retroperitoneal hemorrhage in 4 dogs with rupture of an adrenal gland tumor were determined. All 4 dogs were lethargic and weak with pale mucous membranes on initial examination. Three dogs did not have any history of clinical signs of hyperadrenocorticism or pheochromocytoma prior to examination. In 3 of the dogs, a mass in the area of the adrenal gland was identified with ultrasonography prior to surgery. All dogs developed ventricular premature contractions before or during anesthesia. Three dogs survived adrenalectomy; 1 dog was euthanatized during surgery because of an inability to achieve adequate hemostasis. The remaining 3 dogs all survived more than 5 months after surgery; 1 was euthanatized 9 months after surgery because of rupture of a hepatic mass. On the basis of these results, we suggest that hemodynamic stabilization followed by adrenalectomy is the treatment of choice for dogs with nontraumatic rupture of an adrenal gland tumor and resulting life threatening hemorrhage. PMID- 11497047 TI - Owner survey of headshaking in horses. AB - OBJECTIVE: To determine signalment, history, clinical signs, duration, seasonality, and response to various treatments reported by owners for headshaking in horses. DESIGN: Owner survey. ANIMALS: 109 horses with headshaking. PROCEDURE: Owners of affected horses completed a survey questionnaire. RESULTS: 78 affected horses were geldings, 29 were mares, and 2 were stallions. Mean age of onset was 9 years. Headshaking in 64 horses had a seasonal component, and for most horses, headshaking began in spring and ceased in late summer or fall. The most common clinical signs were shaking the head in a vertical plane, acting like an insect was flying up the nostril, snorting excessively, rubbing the muzzle on objects, having an anxious expression while headshaking, worsening of clinical signs with exposure to sunlight, and improvement of clinical signs at night. Treatment with antihistamines, nonsteroidal anti-inflammatory drugs, corticosteroids, antimicrobials, fly control, chiropractic, and acupuncture had limited success. Sixty-one horses had been treated with cyproheptadine; 43 had moderate to substantial improvement. CONCLUSIONS AND CLINICAL RELEVANCE: Headshaking may have many causes. A large subset of horses have similar clinical signs including shaking the head in a vertical plane, acting as if an insect were flying up the nostrils, and rubbing the muzzle on objects. Seasonality and worsening of clinical signs with exposure to light are also common features of this syndrome. Geldings and Thoroughbreds appear to be overrepresented. Cyproheptadine treatment was beneficial in more than two thirds of treated horses. PMID- 11497048 TI - Marsupialization and iodine sclerotherapy of a branchial cyst in a horse. AB - A 6-month-old Morgan colt was evaluated because of a 10-cm right-sided retropharyngeal swelling. The swelling was soft and moveable on examination, and palpation did not elicit signs of pain. Radiography revealed a large space occupying mass ventral to the second cervical vertebra; ultrasonography revealed an anechoic fluid-filled structure with a well-defined hyperechoic capsule. Fine needle aspiration yielded a viscous amber fluid. Cytologic evaluation indicated that the fluid was an exudate; anaerobic and aerobic bacterial culture did not yield any growth. Histologic examination of a portion of the cyst capsule revealed a connective tissue wall lined by pseudostratified columnar to cuboidal epithelium, consistent with a branchial cyst. The cyst wall was marsupialized to the skin, and iodine sclerotherapy was performed twice daily for 14 days, at which time forceps were introduced into the cyst and the cyst lining was removed. The site was allowed to heal by second intention, but 10 days later, the swelling recurred. An incision was made over the previous marsupialization site, and residual remnants of the cauterized cyst lining were removed with a forceps. The foal did not have any other complications during the subsequent 2 years. Branchial arch cysts are uncommon embryonic anomalies of horses, mice, cats, dogs, and cattle. Results suggest that marsupialization and iodine sclerotherapy may be a viable alternative to surgical excision in horses with branchial cysts; however, the entire cyst lining must be removed at the completion of sclerotherapy to prevent recurrence and abscess formation. PMID- 11497049 TI - Osteomyelitis of the sustentaculum tali in horses: 10 cases (1992-1998). AB - OBJECTIVE: To determine outcome of horses with osteomyelitis of the sustentaculum tali (ST), with or without associated tarsal sheath tenosynovitis, following surgical debridement and lavage. DESIGN: Retrospective study. ANIMALS: 10 horses in which a diagnosis of osteomyelitis of the ST had been made on the basis of history, physical examination findings, and results of radiography. PROCEDURE: Information on results of diagnostic testing, surgical findings, postoperative treatment, and short-term outcome was obtained from the medical records. Long term follow-up information was obtained through reevaluation of horses at the teaching hospital and telephone conversations with referring veterinarians, owners, and trainers. RESULTS: Treatment consisted of surgical debridement, intra and postoperative lavage, and long-term antimicrobial and anti-inflammatory treatment. Eight horses had evidence of involvement of the tarsal sheath. One horse was euthanatized after surgery because of a lack of response to treatment; the other 9 were discharged from the hospital. Severity of lameness had improved, but all still had grade-1 or -2 lameness at the time of discharge. One horse was euthanatized after discharge because of contralateral hind limb laminitis, and another horse was lost to follow-up. Of the remaining 7 horses, 6 returned to their previous use, and 1 was sound but retired for breeding for unrelated reasons. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that horses with osteomyelitis of the ST, with or without concomitant tarsal sheath tenosynovitis, can have an excellent to good outcome and may return to their previous use after surgical debridement of affected tissues and lavage of the tarsal sheath. PMID- 11497050 TI - Tissue disposition and depletion of penicillin G after oral administration with milk in unweaned dairy calves. AB - OBJECTIVE: To determine tissue depletion of penicillin G in calves after oral ingestion with milk replacer and estimate a withdrawal period. DESIGN: Longitudinal controlled trial. ANIMALS: 26 Holstein calves. PROCEDURE: Once daily, 24 calves were fed milk replacer containing procaine penicillin G (0.68 mg/kg [0.31 mg/lb] of body weight); 2 calves served as controls. After 1 feeding, 12 calves were euthanatized in groups of 3 each 4, 6.5, 9.5, and 13 hours after feeding. After 14 days, 12 calves were euthanatized in groups of 3 each 4, 6.5, 9.5, and 13 hours after the final feeding. Concentrations of penicillin G were determined in tissues, blood, and urine by use of high-performance liquid chromatography. RESULTS: Penicillin G was not detected in muscle samples of treated calves. The highest concentrations of penicillin G in plasma, kidney, and liver were 13 ng/ml, 92 ng/g, and 142 ng/g, respectively. Thirteen carcasses had violative drug residues; 12 had violative residues in the liver only, and 1 had violative residues in the liver and kidney. A 21-hour withdrawal period was estimated. CONCLUSIONS AND CLINICAL RELEVANCE: Liver had the highest concentration of penicillin G and was most likely to have violative residues. Feeding calves milk containing penicillin G has the potential to cause violative drug residues in tissues. It is recommended to observe an appropriate withdrawal time prior to slaughter if calves are fed milk from cows treated with penicillin G. PMID- 11497051 TI - Effect of maternal antibodies on induction and persistence of vaccine-induced immune responses against bovine viral diarrhea virus type II in young calves. AB - OBJECTIVE: To determine the effect of maternally derived antibodies on induction of protective immune responses against bovine viral diarrhea virus (BVDV) type II in young calves vaccinated with a modified-live bovine viral diarrhea virus (BVDV) type I vaccine. DESIGN: Blinded controlled challenge study. ANIMALS: 24 neonatal Holstein and Holstein-cross calves that were deprived of maternal colostrum and fed pooled colostrum that contained a high concentration of (n = 6) or no (18) antibodies to BVDV. PROCEDURE: At 10 to 14 days of age, 6 seropositive and 6 seronegative calves were given a combination vaccine containing modified live BVDV type I. All calves were kept in isolation for 4.5 months. Six calves of the remaining 12 untreated calves were vaccinated with the same combination vaccine at approximately 4 months of age. Three weeks later, all calves were challenged intranasally with a virulent BVDV type II. RESULTS: Seronegative unvaccinated calves and seropositive calves that were vaccinated at 2 weeks of age developed severe disease, and 4 calves in each of these groups required euthanasia. Seronegative calves that were vaccinated at 2 weeks or 4 months of age developed only mild or no clinical signs of disease. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that a single dose of a modified-live BVDV type-I vaccine given at 10 to 14 days of age can protect susceptible young calves from virulent BVDV type II infection for at least 4 months, but high concentrations of BVDV-specific maternally derived antibodies can block the induction of the response. PMID- 11497052 TI - Serum immunoglobulin G concentrations in calves fed fresh and frozen colostrum. AB - OBJECTIVE: To determine whether serum IgG concentrations in neonatal calves are adversely affected by short-term frozen storage of colostrum. DESIGN: Prospective study. SAMPLE POPULATION: Experiment 1 consisted of 10 pairs of Holstein calves (n = 20) fed matched aliquots of either fresh (n = 10) or frozen and thawed (10) colostrum. In experiment 2, 26 Holstein calves were fed either fresh (n = 13) or frozen and thawed (n = 13) colostrum. PROCEDURE: Experiment 1 consisted of calves resulting from observed parturitions; calves were randomly assigned to treatment groups (fresh or frozen and thawed colostrum) in pairs. Calves were fed 4 L aliquots of colostrum via oroesophageal intubation at 3 hours of age. Serum IgG concentrations at 2 days of age were compared between the 2 groups by use of a paired t-test. Experiment 2 consisted of calves resulting from observed parturitions; calves were randomly assigned to treatment groups (fresh or frozen and thawed colostrum). Calves were fed 4 L aliquots of colostrum via oroesophageal intubation at 3 hours of age. Regression analysis was used to determine whether calf serum IgG concentration was a function of colostral IgG concentration and colostrum storage group. RESULTS: Significant differences were not observed between the 2 groups in experiment 1. No significant relationship was observed between colostrum storage group and serum IgG concentration in experiment 2. The model that best predicted serum IgG concentrations accounted for 20% of the variability in serum IgG concentration. CONCLUSION AND CLINICAL RELEVANCE: Frozen colostrum is an adequate source of IgG for calves. PMID- 11497053 TI - Monocular and binocular distance cues: insights from visual form agnosia I (of III). AB - The human nervous system constructs a Euclidean representation of near (personal) space by combining multiple sources of information (cues). We investigated the cues used for the representation of personal space in a patient with visual form agnosia (DF). Our results indicated that DF relies predominantly on binocular vergence information when determining the distance of a target despite the presence of other (retinal) cues. Notably, DF was able to construct an Euclidean representation of personal space from vergence alone. This finding supports previous assertions that vergence provides the nervous system with veridical information for the construction of personal space. The results from the current study, together with those of others, suggest that: (i) the ventral stream is responsible for extracting depth and distance information from "monocular" retinal cues (i.e. from shading, texture, perspective) and (ii) the dorsal stream has access to binocular information (from horizontal image disparities and vergence). These results also indicate that DF was not able to use size information to gauge target distance, suggesting that intact temporal cortex is necessary for "learned size" to influence distance processing. Our findings further suggest that in neurologically intact humans, object information extracted in the ventral pathway is combined with the products of dorsal stream processing for guiding prehension. Finally, we studied the "size-distance paradox" in visual form agnosia in order to explore the cognitive use of size information. The results of this experiment were consistent with a previous suggestion that the paradox is a cognitive phenomenon. PMID- 11497054 TI - Vertical gaze angle as a distance cue for programming reaching: insights from visual form agnosia II (of III). AB - It has been shown that a patient with visual form agnosia (DF) relies predominantly on vergence information when gauging target distance in an open loop pointing task. This finding suggested that the programming of prehension might be severely disrupted if DF viewed target objects through ophthalmic prisms. An initial experiment showed that this prediction was not upheld; DF was able to programme reasonably accurate movements to objects located on a tabletop despite large changes in vergence angle. A second experiment, however, showed that placing the target objects at eye height whilst manipulating vergence angle caused gross disruption to prehension, with DF mis-programming the reach component in a predictable manner. Notably, the evidence for DF's reliance on vergence distance information was obtained in a task where the targets were viewed at eye height. These experiments indicate that DF uses vertical gaze angle to gauge target distance in normal prehension and suggest that this extra-retinal cue may be a useful source of distance information for the human nervous system, especially where pictorial cues are impoverished. PMID- 11497055 TI - The role of size and binocular information in guiding reaching: insights from virtual reality and visual form agnosia III (of III). AB - Reaching out to grasp an object requires information about the size of the object and the distance between the object and the body. We used a virtual reality system with a control population and a patient with visual form agnosia (DF) in order to explore the use of binocular information and size cues in prehension. The experiments consisted of a perceptual matching task in addition to a prehension task. In the prehension task, control participants modified their reach distance in response to step changes in vergence in the absence of any clear reference for relative disparity. Their reach distance was unaffected by equivalent step changes in size, even though they used this information to modify grasp and showed a size bias in a distance matching task. Notably, DF showed the same pattern of results as the controls but was far more sensitive to step changes in vergence. This finding complements previous research suggesting that DF relies predominantly on vergence information when gauging target distance. The results from the perceptual matching tasks confirmed previous findings suggesting that DF is unable to make use of size information for perceptual matching, including distance comparisons. These data are discussed with regard to the properties of the pathways subserving the two visual cortical processing streams. PMID- 11497056 TI - Guiding contact by coupling the taus of gaps. AB - Animals control contact with surfaces when locomoting, catching prey, etc. This requires sensorily guiding the rate of closure of gaps between effectors such as the hands, feet or jaws and destinations such as a ball, the ground and a prey. Control is generally rapid, reliable and robust, even with small nervous systems: the sensorimotor processes are therefore probably rather simple. We tested a hypothesis, based on general tau theory, that closing two gaps simultaneously, as required in many actions, might be achieved simply by keeping the taus of the gaps coupled in constant ratio. tau of a changing gap is defined as the time-to closure of the gap at the current closure-rate. General tau theory shows that tau of a gap could, in principle, be directly sensed without needing to sense either the gap size or its rate of closure. In our experiment, subjects moved an effector (computer cursor) to a destination zone indicated on the computer monitor, to stop in the zone just as a moving target cursor reached it. The results indicated the subjects achieved the task by keeping tau of the gap between effector and target coupled to tau of the gap between the effector and the destination zone. Evidence of tau-coupling has also been found, for example, in bats guiding landing using echolocation. Thus, it appears that a sensorimotor process used by different species for coordinating the closure of two or more gaps between effectors and destinations entails constantly sensing the taus of the gaps and moving so as to keep the taus coupled in constant ratio. PMID- 11497057 TI - Vision influences tactile perception at body sites that cannot be viewed directly. AB - Previous research has demonstrated that vision of a body site, without proprioceptive orienting of eye and head to that site, could affect tactile perception. The body site viewed was the hand, which can be seen directly under normal viewing conditions. The current research asked three further questions: First, can vision similarly affect tactile perception at a body site that cannot normally be viewed directly such as the face or neck? Second, does prior experience of seeing a body site, such as occurs when viewing the face in mirrors, produce larger effects of viewing than body sites rarely seen such as the back of the neck? And third, how quickly can visual information affect tactile target detection? We observe that: detection of tactile targets at these body sites was influenced by whether or not they were viewed, this effect was greater when viewing the more familiar site of the face than that of the neck, and significant effects were observed when the stimulus onset asynchrony between visual display and tactile target was as little as 200 ms. PMID- 11497058 TI - Inhibition of biceps brachii muscle motor area by painful heat stimulation of the skin. AB - ract This study aimed to evaluate whether painful cutaneous stimuli can affect specifically the excitability of the arm proximal muscle motor area. The motor evoked potentials (MEPs), recorded from the right biceps brachii muscle after either transcranial magnetic or electrical anodal stimulation of the left primary motor (MI) cortex, were conditioned by painful CO2 laser stimuli delivered either on the right hand dorsum or on the lateral surface of the right arm. Painful CO2 laser stimuli delivered on the hand skin reduced significantly the amplitude of MEPs evoked by the transcranial magnetic stimulation of the contralateral MI area, while the MEP amplitude was not significantly modified by CO2 laser pulses delivered on the arm skin. The inhibitory effect followed the arrival of the nociceptive inputs to the cerebral cortex. The amplitude of MEPs evoked by anodal electrical stimulation of the motor cortex was not decreased by conditioning painful stimuli delivered on the hand dorsum. Since the magnetic stimulation led to transynaptic activation of pyramidal neurons, while the anodal stimulation activated directly corticospinal axons, our findings suggest that CO2 laser pulses delivered on the hand are able to inhibit the arm proximal muscle motor area. PMID- 11497059 TI - Chameleons have independent eye movements but synchronise both eyes during saccadic prey tracking. AB - The movements of both eyes and the head were recorded with search coils in unrestrained, freely moving chameleons. As a main result I found that the generation of saccades in the left and the right eye was either independent from each other or was highly correlated according to the behavioural situation. When no prey item was fixated, disconjugate saccades were observed which was in accordance with earlier observations in chameleons. During prey tracking the chameleons switched to a different oculomotor behaviour and pursued the moving prey with synchronous saccades. At higher target velocities, the tracking movement of the head was also saccadic and was synchronised with the two eyes. Binocular coupling affected only the timing of the saccades but not the metrics: the amplitudes of the synchronous saccades were usually different in the two eyes. These observations suggest the existence of two independent premotor neuronal circuits for left and right eye saccadic motor control in the chameleon. Binocular coupling in prey-tracking chameleons is probably achieved by neuronal coupling of these premotor circuits during eye-head coordination. The ability to switch between synchronous and uncoupled saccadic eye movements has not been described for any other vertebrate. This unique ability of the chameleon may help to understand the organisation of the oculomotor system of other vertebrates since evidence for separate left eye and right eye saccade generation and position control has recently also been reported in primates. PMID- 11497060 TI - Colour-related oscillations in the striate cortex of awake monkeys: "reverse" observations. AB - Gamma oscillations of 30-70 Hz in local electroencephalograms (EEGs) were observed in primary visual cortex (V1) of monkeys when they viewed coloured stimuli under conditions which were not part of a training paradigm. No oscillatory modulations were detected in simultaneously recorded spike trains, although the latter responded to the stimuli. The oscillations had much longer latencies than the spike responses. Proceeding in a "reverse" manner, the oscillations were taken as primary elements of the analyses, and relationships to the accompanying behaviour were sought. Besides colour stimulation, the oscillations were found to be related to a form of annoyance. Only two of four monkeys showed the phenomena. We conclude that the oscillations had a central origin. PMID- 11497061 TI - Quantitative changes in gene expression of glutamate receptor subunits/subtypes in the vestibular nucleus, inferior olive and flocculus before and following unilateral labyrinthectomy in the rat: real-time quantitative PCR method. AB - Spontaneous recovery from the oculomotor and postural symptoms of unilateral labyrinthectomy (UL) is known as vestibular compensation, which is a useful model for investigation of the mechanisms of lesion-induced CNS plasticity. In the present study, to elucidate the molecular biological basis of vestibular compensation, we investigated changes in the mRNA expression of glutamate receptor subunit/subtypes in the rat central vestibular system, including the vestibular nucleus complex (VNC), inferior olive (IO), and cerebellar flocculus following UL, using a real-time quantitative polymerase chain reaction (PCR) method. In normal control animals, regional differences in the expression of several glutamate receptor subunit/subtypes, e.g., NR1 and NR2A subunits of the N methyl-D-aspartic acid (NMDA) receptor, GluR2 and KA2 subtypes of non-NMDA receptors, and mGluR1 and mGluR7 metabotropic glutamate receptors, were consistent with previous results from studies using in situ hybridization histochemistry, suggesting that the real-time quantitative PCR method was a reliable procedure for evaluation of changes in mRNA expression. In the vestibular nucleus complex, NR2A, GluR2 and mGluR7 mRNA were ipsilaterally downregulated by 6 h following UL (P<0.05, P<0.05 and P<0.01, respectively). In the inferior olive, no changes in gene expression were observed. In the ipsilateral flocculus, KA2 mRNA expression was increased by 50 h post-UL (P<0.05). However, in the contralateral flocculus, mGluR1 mRNA was downregulated by 6 h post-UL (P<0.005). Both the increase in KA2 mRNA expression in the ipsilateral flocculus and the decrease in mGluR1 mRNA expression in the contralateral flocculus may have had the effect of reducing Purkinje cell inhibition of ipsilateral VNC neurons, thereby contributing to the rebalancing of spontaneous resting activity between the ipsilateral and contralateral VNCs. It is suggested that such changes in the activities of the floccular-VNC pathways may be important to the vestibular compensation process. PMID- 11497062 TI - Adaptive plasticity of head movement propensity. AB - Individual humans exhibit differing propensity to move the head in association with saccadic shifts in gaze. We assessed whether this tendency can be modified in normal subjects by either reducing neck mobility with a cervical collar or restricting the field of view using aperture spectacles. We quantified head movement propensity in terms of the range of orbital eccentricity within which the eyes are customarily maintained (customary ocular motor range), and the range of final eye-in-head eccentricity for which a planned saccade is likely to be executed without a concomitant head movement (eye-only range). Three subjects wore rigid collars during waking hours for periods of up to 9 days. We measured customary and eye-only ranges with the collar removed, at various times during the adaptation and recovery periods. Collar adaptation reduced head movement propensity in all three subjects, increasing the average customary ocular range from 27.6 +/- 8.9 degrees (mean +/- SD) to 66.1 +/- 4.5 degrees and the eye-only range from 24.6 +/- 17.0 degrees to 67.6 +/- 7.4 degrees. In two subjects the modifications persisted for weeks following final collar removal. In parallel with the reduction in head movement propensity, all subjects improved in their ability to maintain eccentric gaze, suggesting that neck restriction led to effects at the level of the brainstem. Three subjects were adapted to spectacles, masked to restrict the field of view to approximately 20 degrees. The aperture spectacles were worn for periods of up to 9 days. When tested without the apertures, one subject exhibited a definite increase in head movement propensity; in the other two, the data were equivocal, indicating either a small increase in head movement propensity or no effect. Averaged across subjects, customary ocular motor range decreased from 35.1 +/- 12.8 degrees to 25.4 +/- 13.4 degrees and eye only range decreased from 35.1 +/- 7.5 degrees to 23.0 +/- 4.0 degrees. The marked difference in the magnitudes of collar- and spectacle-induced changes suggests that the responses to the two restrictive appliances are mediated by different mechanisms. Collar adaptation may involve parametric modulation of circuits mediating reflex recruitment of head movements, while aperture adaptation may primarily reflect substitution of an alternative mode of head control triggered by the presence of the restricted field of view, with only minor parametric modulation of the underlying head recruitment circuit. The enduring effects of restricting neck mobility upon head movement tendencies may relate to the common clinical association between neck injury and persistent dysequilibrium. PMID- 11497063 TI - Jaw muscle response to stimulation of type II somatosensory afferents of limbs in the rat. AB - Convergence of various afferent inputs onto brainstem neurones may play an important role in the regulation of trigeminal motor activity. In particular, previous studies suggest that, besides sensory inputs arising from the orofacial region, extratrigeminal information may modulate jaw muscle function. In the present study the actions exerted on masseter and digastric muscles by the activation of somatosensory afferents coming from fore- and hind limbs were examined. The electromyographic activity (EMG) of masseter and digastric muscles was recorded in 20 anaesthetised rats, and EMG responses to single and paired electrical stimulation of common radial and sciatic nerves, at a threshold intensity for the activation of group II afferent fibres, were studied. The stimulation induced an excitatory response in both masseter and digastric muscles bilaterally. Ipsiand contralateral radial nerve stimulation evoked masseter responses at latencies of 13.8 +/- 2.4 ms and of 18.0 +/- 2.6 ms, respectively, and digastric responses 1.6 +/- 0.4 ms later. Ipsi- and contralateral sciatic nerve stimulation elicited masseter responses at latencies of 21.4 +/- 2.6 ms and of 23.3 +/- 2.0 ms, respectively, and digastric responses 2.0 +/- 0.2 ms later. The same masseter and digastric motor units were excited by both radial and sciatic nerve stimulation; this suggests a convergence of somatosensory inputs arising from fore- and hind limbs on the same pool of masseter and digastric motoneurones. Paired stimulation of the two nerves did not induce any summation of the responses; this finding suggests that the two inputs, reaching a common relay station, could give rise either to occlusion or to inhibitory interactions. Spinotrigeminal relationship evidenced in this study may be involved in the coordination of jaw and limb movements. PMID- 11497064 TI - Delayed loss of spinal motoneurons after peripheral nerve injury in adult rats: a quantitative morphological study. AB - The existence of retrograde cell death in sensory dorsal root ganglion (DRG) cells after peripheral nerve injury is well established. However, with respect to retrograde motoneuron death after peripheral nerve injury, available data are conflicting. This may partly be due to the cell counting techniques used. In the present study, quantitative morphometric methods have been used to analyse retrograde motoneuron death induced by spinal nerve injury in adult rats. For comparison, DRG cells were also included in the study. The C7 spinal nerve was transected about 10 mm distal to the DRG and exposed to the fluorescent tracer fast blue in order to retrogradely label the spinal motoneurons and DRG cells of the C7 segment. At 1-16 weeks postoperatively, the nuclei of fast-blue-labelled C7 motoneurons and DRG cells were counted in consecutive 50-microm-thick serial sections. For comparison, the physical disector technique and measurements of neuronal density were also used to calculate motoneuron number. The counts of fast-blue-labelled motoneurons revealed a delayed motoneuron loss amounting to 21% and 31% after 8 and 16 weeks, respectively (P<0.001). The remaining motoneurons exhibited 20% (P<0.05) soma atrophy. Using the physical disector technique, the motoneuron loss was 23% (P<0.001) after 16 weeks. Calculations of neuronal density in Nissl-stained sections failed to reveal any motoneuron loss, although after correction for shrinkage of the ventral horn a 14% (P<0.001) motoneuron loss was found. The fast-blue-labelled DRG neurons displayed 51% (P<0.001) cell loss after 16 weeks, and the remaining cells showed 22% (P<0.001) soma atrophy. In summary, cervical spinal nerve injury induces retrograde degeneration of both motoneurons and DRG cells. However, to demonstrate the motoneuron loss adequate techniques for cell counts have to be employed. PMID- 11497065 TI - Development of postural adaptation to arm raising. AB - We studied the development of the coordination between posture and movement by analyzing the shifts of the center of pressure (CoP) associated with arm raising. Three groups of children aged 3-5 years, 6-8 years, and 9-10 years and an adult group were tested. The subjects were required to raise their arms to the horizontal position while standing still, with their hands free or loaded (5% of the body weight). The arm movements were recorded by a TV-image processor, and the changes in position of the CoP were measured by a force platform and analyzed before, during, and after the arm movement. The data show that the CoP moved forward during arm raising, that additional load induced a greater shift in all age groups, and that the relative amplitude of the shift decreased with age. The greatest changes occurred between ages 3-5 years and 6-8 years. The pre- and postmovement CoP shift suggests qualitative changes in the postural adaptation to movement between these two age groups: the anticipatory postural adjustments moved from a supporting function to a compensatory function, yielding an increasing functional convergence between the feedforward and the feedback modes of postural control, and an increasing rapidness in recovering postural stability after arm movement. The postural behavior shown by the 9- to 10-year-old children and by the adults in the arms-free condition suggests an increased tolerance to unbalance when postural oscillations do not jeopardize static equilibrium. PMID- 11497067 TI - Mechanisms for sensorimotor adaptation to rotated visual input. AB - Using the multiple-exposure approach, we investigated sensorimotor adaptation by exposing human subjects to different angles of visual rotation in a tracking task. Generally, the tracking error was high at the onset of the visual rotation and gradually declined towards the baseline level during the exposure period. In experiment A, we confirmed that the initial tracking error increases more than proportionally with the angle of rotation. In experiment C, we were unable to confirm intermanual transfer, and attribute this discrepancy with previous literature to details of the experimental tasks. In our main experiment, B, we found that pre-exposure to 45 degrees or 60 degrees of visual rotation facilitated the subsequent adaptation to a 90 degrees rotation, with the facilitatory effect being more pronounced following the 60 degrees rotation. We interpret this finding as evidence that adaptation is achieved by a gradual process, which progresses from small angles of output transformation through intermediate values up to the prescribed angle of rotation. PMID- 11497066 TI - Predictive elements in ocular interception and tracking of a moving target by untrained cats. AB - We presented a mechanical target moving at constant velocity to awake, nontrained, head-restrained cats, in order to study how naive animals pursue objects moving at a high speed with their gaze. Eye movements were recorded while the target was moving in different directions at a constant velocity (20-80 degrees/s) through the center of the visual field. We observed two oculomotor strategies: cats either made an interception saccade (IS) toward the target but opposite to its motion, or tracked it in the direction of motion. They used the interception strategy more frequently when the gaze position error at the onset of target motion was large, and the tracking strategy when it was small. Interception was always achieved by single saccades, which were faster than tracking saccades (TS). During tracking, cats generated sequences of two to six saccades separated by "smooth" eye movements. Tracking quality varied considerably from trial to trial. When the level of motivation was high, cats would track the target at 80 degrees/s over up to 75% of the oculomotor range, with relatively small position errors. We compared ISs and TSs with respect to their metric properties and timing. The amplitudes of ISs positively correlated with position error existing 100 ms before saccade onset, but saccade vectors were directed to a point ahead of the target along the target's track. We conclude that, in programming the ISs, target motion is used to predict the future target position so as to assure a spatial lead of the gaze at the saccade end, instead of attempting a precise capture of the target. The amplitude of TSs did not depend on preceding position errors. TSs were usually small at the onset of the first saccade, as if cats would wait till the target arrived near the line of sight. A majority of primary TSs were initiated before the target arrived near the direction of gaze. Thus they had a direction, opposite to the position error sampled 100 ms before the saccade, but the same as the direction of target motion. Prediction of the future target position from its velocity vector should therefore contribute to the programming of TSs. In addition, we observed that TSs were faster when they were initiated with a spatial lag relative to the target and they were slower if there was a spatial lead or target velocity was reduced. Such a modulation appears to be analogous to the predictive correction of the saccade amplitude during smooth pursuit in primates. Considering strong visual motion sensitivity and motor properties of output neurons of the superior colliculus, it is likely that, in cats, the colliculus makes a major contribution to the integration of eye movement-related and target motion-related signals. PMID- 11497068 TI - Dog days [editorial]. PMID- 11497069 TI - The Human Genome Project: ethical and legal considerations for neuroscience nurses. AB - The Human Genome Project has nearly completed the identification of the human genome. The medical, social, and legal implications are numerous. Neuroscience nurses have the unique opportunity to be an integral part of the genetics age because of the many genetic-based neuroscience diseases. PMID- 11497070 TI - Alcohol, head injury, and pulmonary complications. AB - The purpose of this study was to determine the difference in rates of pulmonary complications (e.g., aspiration, pneumonia) in head-injured patients with and without concomitant alcohol intoxication. The records of 98 consecutive patients admitted over a 1-year period to a Level I Trauma Center were reviewed. The patients were grouped into three subsets: acutely intoxicated (n = 26), acutely intoxicated with a diagnosis of chronic alcoholism (n = 14), and non-intoxicated (n = 58). Alcohol intoxication was defined as a blood alcohol level (BAL) > or = 0.08 mg/dl. Admission BALs and Glasgow Coma Scale (GCS) scores were tabulated at admission. Frequency of arterial blood gas (ABG) measurements, need for an artificial airway/mechanical ventilation, and length of stay (LOS) were analyzed by using one-way analysis of variance. Intergroup differences in breath sounds were compared by using the nonparametric Kruskall-Wallis technique. We found no statistical difference between groups in terms of pulmonary sequelae despite the remarkably high BALs observed in the study groups. Similarly, there was no statistically greater LOS in the groups with alcohol intoxication than in alcohol free cohorts. Despite a great deal of BAL science research to support our hypothesis, we failed to demonstrate a significantly higher rate of pulmonary problems in inebriated individuals with head injuries. We found that our strict exclusion criteria (no concomitant chest, abdominal, or pelvic trauma) limited the sample to only those patients without significant intracranial bleeding, whereas most complications in blood alcohol neuroscience research have been associated with much larger mass lesions (e.g., epidural or subdural hematomas). In addition, we found the characterizations of patients as chronically alcoholic were cumbersome and inaccurate in many cases. Future research should allow for a greater range of concomitant injuries that might suggest a positive or negative relationship to acute intoxication. PMID- 11497071 TI - Infant neurologic assessment. AB - Infant neurologic assessment reflects the ongoing maturation of the central nervous system. Traditional approaches to assessment cannot be used. Key factors are accurate observation and flexibility in obtaining the data. A case example using a 4-month-old infant illustrates specific approaches to assessment. PMID- 11497072 TI - Assessing functional ability in persons with dementia: using family caregivers as informants. AB - More than one in five community-dwelling older individuals is unwilling or unable to provide information on functional abilities. In such situations the standard procedure is to augment self-reports with those of family members or other close informants. However, when these reports differ, it often is difficult to determine whether the older individual is overly optimistic about his or her functional abilities or the family informant is unduly pessimistic. This article explores factors that influence family caregiver assessments of functional abilities in older individuals with some degree of cognitive loss or impairment and presents suggestions for enhancing the accuracy and dependability of functional assessments by family informants. PMID- 11497073 TI - Respiratory failure in the neurological patient: the diagnosis of neurogenic pulmonary edema. AB - Neurogenic pulmonary edema (NPE) is a potential complication of a central nervous system (CNS) insult such as intracranial hemorrhage, uncontrolled generalized seizures, head trauma, tumors, and neurosurgical procedures. The proposed etiology is massive sympathetic discharge following a CNS event. The pathogenesis is not completely understood. However, there are two theories on how NPE occurs: the blast theory and the permeability defect theory. There is evidence for both theories, and NPE is probably the result of a combination of the two. The treatment is mainly supportive with the use of mechanical ventilation and alpha adrenergic blocking agents while managing increased intracranial pressure. A thorough understanding of the pathophysiological mechanisms behind the development of NPE aids in the management of these patients to prevent further complications. PMID- 11497074 TI - Sleep, memory, and learning. AB - The relationship of sleep to memory and learning is complex. Sleep affects memory, and memory must be present for learning to occur. A number of studies have been conducted to increase our understanding of their relationship. In addition to the numerous scientific investigations of each concept separately, sleep, memory, and learning have been studied together to determine (a) the effect of sleep on memory and learning, (b) the effect of sleep deprivation in general on memory and learning, (c) the effect of rapid eye movement (REM) sleep deprivation on memory and learning, (d) the effect of memory and learning on REM sleep, and (e) the effect of non-REM sleep loss on memory and learning. Neuroanatomic correlates have been pursued as well with most attention to the hippocampus. Despite considerable efforts to date, many of the studies reveal contradictory or inconclusive findings. Much remains unknown, and additional work is needed. Implications for nursing include those that have a direct effect on the patient, the nurse, and nursing science. PMID- 11497075 TI - Effect size: utility and application in neuroscience nursing. AB - Effect size is used in single studies to complement inferential statistical analysis. It also is used to compare and synthesize data across studies in meta analysis. Effect size provides information about clinical importance of differences found in research observations. For researchers, effect size is useful in establishing support for the theoretical framework of a story and in planning for future research. Application of effect size to data in three studies about the effects of backrest position on ICP and CPP in brain-injured adults illustrates the utility of effect size analysis in neuroscience nursing research. PMID- 11497076 TI - Relationships between the presence of anti-Tat antibody, DNA and RNA viral load. AB - The possible relationships between the intensity of humoral response to full length Tat protein, the amount of proviral DNA reservoir in peripheral blood mononuclear cells and RNA viral load were analyzed in plasma samples obtained from a group of HIV-1 seropositive subjects, who never received any antiretroviral therapy. All HIV-1 patients showed detectable levels of serum IgG to full-length Tat by immunoenzymatic assay. We found a higher percentage of HIV 1 seropositive subjects with low levels of antibody in the presence of barely detectable proviral DNA copies (< or =10 copies/1.5x10(5) PBMCs) and a high anti Tat antibody response accompanied by variable (from >10(1) to > or =10(3) copies/1.5x10(5) PBMCs) levels of DNA load (p=0.011). Moreover, an inverse relationship between anti-Tat antibody titers and HIV-1 RNA viral load was demonstrated HIV-1 seropositive patients. In HIV-1-infected patients, a strong humoral immune response against HIV-1 transactivating Tat protein, able to down modulate viral replication in peripheral blood, does not seem to inhibit the number of proviral DNA molecules in peripheral blood mononuclear cells. Even though our data strongly confirm the "positive" role of anti-Tat antibody on viral replication, the persistence of significant amount of DNA viral load in peripheral blood mononuclear cells, despite high level of anti Tat antibody, suggests a more cautious approach to HIV-1 Tat-containing vaccines, able to stimulate an immune specific response to transactivating Tat protein sufficient in inhibiting circulating virus, but not completely efficient in decreasing proviral DNA integration. PMID- 11497077 TI - Isolation of coxsackievirus B5 from pigs. AB - A cytopathic virus was isolated from young pigs suffering from severe diarrhea in Okinawa, Japan in 1986. The disease was highly contagious among young pigs. The physico-chemical properties of the virus indicated an enterovirus, but, no of serological relationship was detected with reference strains of porcine enteroviruses. With the aid of Genbank for genomic sequence data, RT-PCR and hybridization method was performed. The viral isolate was identified as the Coxsackievirus (CV) B5 of human enterovirus. This is the first report of the isolation of CV B5 from pigs in Japan. PMID- 11497078 TI - Quantitative PCR in EBV-infected renal transplant patients. AB - In this study we investigated the levels of Epstein Barr virus (EBV) DNA by quantitative polymerase chain reaction (Q-PCR) in serum, whole blood and peripheral blood mononuclear cells (PBMC) from anti-EA IgG seropositive or anti EA IgG seronegative EBV infected renal transplant recipients. We compared serological data with the viral load to monitor the risk of developing post transplant lymphoproliferative disorders (PTLD). All patients were asymptomatic and none of them developed PTLD at the time of the study. EBV DNA quantitation for each patient varied in whole blood and PBMC samples probably due to different numbers of mononuclear cells present in samples from which DNA was extracted (whole blood vs. purified PBMC). In 92% of the serum samples EBV DNA was undetectable probably due to absence of free genomes since the number of DNA copies detected in samples from whole blood and PBMC does not reach very high levels. The correlation between the presence of EA-antibody, considered serological evidence of EBV reactivation, and the viral load showed that 60% of EA-positive patients had quantifiable EBV DNA, whereas in 40% of EA-positive patients EBV DNA was undetectable, showing serological reactivity but no viral replication. Of the remaining EA-negative patients, EBV DNA could be detected in 71% of them, whereas 29% did not show EBV DNA, indicating no EBV replication. In conclusion, our results confirm that the presence of serum IgG anti-EA antibody is not a reliable marker of active EBV infection whereas the evaluation of the viral load in blood samples is a useful diagnostic tool to monitor and to better understand the course of EBV infection in immunocompromised renal transplant patients at risk of developing PTLD. PMID- 11497079 TI - Control of murine cytomegalovirus replication in salivary glands during acute infection is independent of the Fas ligand/Fas system. AB - The course of mouse cytomegalovirus (MCMV) infection was compared between mutant C57BL/6 (B6) mice deficient in either perforin (perf-/-), or perforin, granzyme A and B (perfxgzmAxB-/-), and B6 gld mice lacking functionally active Fas ligand to elucidate the contribution of the two main cytolytic pathways in the early control of MCMV infection. At 15 and 30 days post infection (p.i.) virus titers were elevated in salivary glands of perf-/- and perfxgzmAxB-/-, but almost undetectable in those of mutant gld and C57BL/6 wild-type mice. No virus was detectable in lung and spleen tissues of the mutant or B6 mice at the time points tested. At 15 days p.i., scanty lymphocytic periductal infiltration was seen in salivary glands of perf-/- and perfxgzmAxB-/; these pathological alterations were minimal at 30 days p.i.. In contrast, no pathological alterations were seen in the respective organs of infected B6 and gld mice at the two time points p.i.. At 15 days p.i., reactive follicles were observed in the white pulp of spleen tissues from both mutant and B6 mice, but at 30 days p.i. only in those of mutant mice. No inflammatory responses were seen in the lung tissues of any of the four mouse strains tested. Together with previous observations (Riera et al.. 2000), the results demonstrate that both perforin and granzymes A/B, but not the FasL/Fas system are critical for viral elimination in salivary glands during the acute phase of infection. However, for the long-term control of MCMV infection, neither of the two cytolytic pathways seem to be necessary. PMID- 11497080 TI - Characterization of a canine parvovirus strain isolated from an adult dog. AB - A CPV-2b strain was detected from an adult vaccinated dog, affected with severe gastroenteritis. The faeces of the dog were positive to canine parvovirus by a hemagglutination assay and gave a CPV-2b-like pattern by a hemagglutination inhibition test using monoclonal antibodies. In vitro-cultivation of the virus was difficult and after a few passages on canine and feline cells, the presence of the virus was detectable only by an immunofluorescence assay on the feline cells, since hemagglutinating activity had disappeared. Characterization of the virus, by an indirect immunofluorescence assay with monoclonal antibodies, confirmed the antigenic CPV-2b-like pattern of the nonhemagglutinating virus. PMID- 11497081 TI - Establishment of a swine monocyte cell line. AB - A swine monocyte cell line was established from peripheral blood sample collected from a healthy adult male pig. The cloned cells grow actively in forming monolayers in both glass and plastic cell culture flasks with the growth medium reported previously (Kadoi, 2000) at 36.5 degrees C incubation. The plating efficiency is more than 95%. Densely grown cells in flasks show an epithelioid morphology. The fundamental properties of the cells were examined for cytological definition as monocytes. A positive property detected was guinea pig complement receptor, porcine IgG receptor, non-specific esterase, and acid phosphatase. A significant phagocytic activity proved by the inoculation of Saccharomyces cerevisiae is also one of the characteristics observed in the LPS-activated cells. PMID- 11497082 TI - Leptospira strains kept at the National Centre for Leptospirosis in Rome, Italy. AB - Since the National Centre for Leptospirosis (Department of Bacteriology and Medical Mycology, Istituto Superiore di Sanita, Rome) was established in 1956 by B. Babudieri, efforts have been devoted to identifying new Leptospira isolates and maintaining a collection of strains that today comprises 670 strains, 550 of which have been totally or partially classified, and 120 are still under study. This collection includes 23 serogroups and 156 serovars of pathogenic leptospires, and 32 serogroups and 54 serovars of saprophytic leptospires. The conventional serogroup and serovar identification, mainly based on antigenic relatedness, is tedious and time-consuming, requiring the maintenance of a comprehensive collection of serovar reference strains and the preparation of the corresponding rabbit antisera. Although considerable difficulties are encountered in the classification of leptospires at the serogroup and serovar level, this classification system is essential to obtain information on the epidemiology of leptospirosis in the different geographical areas. Serovar identification has become faster with the introduction of pulsed-field gel electrophoresis (PFGE) of large DNA fragments obtained after digestion of leptospiral DNAs with rare cutting restriction enzymes. This technique has been successfully utilized to discriminate between closely related serovars of the Leptospira interrogans complex. We have recently used PFGE to characterize several Italian leptospiral isolates, confirming that PFGE analysis combined with microscopic agglutination test (MAT) with monoclonal and polyclonal antibodies can be used as an accurate and reliable method to compare and classify leptospires. PMID- 11497083 TI - Detection of Legionella pneumophila at thermal spas. AB - Water samples were collected at three therapeutic thermal spas in the area of Brescia, between February and October 2000: 34.8% of the samples contained Legionella pneumophila; the predominant isolates (30%) belonged to Legionella pneumophila serogroup 1. The microorganism was present in the spa water at high concentrations, generally higher than 10000 cfu/l. The large number of positive Legionella pneumophila samples indicates a potential risk of infection to patients, especially those undergoing inhalation treatment with thermal water, or those using a whirlpool or taking a shower even if, during the study, no clinical cases of Legionnaires' disease were observed. In some inhalators in use we detected Legionella pneumophila: after a treatment to eradicate the microorganism, no sanitary fittings currently show contamination. Thus, in our opinion, they are not sources of infection when they are mantained and serviced properly. Thermal disinfection and service checks at regular intervals are suggested for contaminated systems. PMID- 11497084 TI - Microplankton respiration (ETS) in two areas of the Northern Adriatic (Mediterranean Sea). AB - Respiratory rates in two areas of the North Adriatic Sea were studied in the context of a PRISMA II Project, with the aim of quantifying the oxidative processes occurring in the frontal system caused by strong riverine discharge. In the northern area, higher respiratory rates and seasonal variations occurred with respect to the southern area. On average, respiratory activity sustained the oxidation of 66.9 nM C h(-1) inside the front and 24.4 nM C h(-1) outside the front. In conclusion, organic matter oxidation inside the front resulted 2.7 times higher than outside the front. Oxidative metabolism in the Northern Adriatic Sea appears mainly governed by river inputs. PMID- 11497085 TI - Virulence factors in Vibrios and Aeromonads isolated from seafood. AB - Thirty-one isolates from seafood, identified as Aeromonas hydrophila (7), Aeromonas caviae (11), Vibrio parahaemolyticus (3), Vibrio fluvialis (5), Vibrio alginolytictus (3), Vibrio metschnikovii (1) and Vibrio damsela (1), were tested for possible virulence factors including extracellular hydrolytic enzymes, haemolysins, cytotoxins (VERO and HEp-2 cells) and adherence ability (HEp-2 cells). All the A. hydrophila strains were beta-haemolytic and produced cytotoxins as well as one strain of V. fluvialis. A. hydrophila and A. caviae strains, frequently adhesive, showed both aggregative and diffusive patterns, while five Vibrio strains only (three V. fluvialis, one V. parahaemolyticus and one V. alginolyticus) were adhesive with an aggregative pattern. PMID- 11497086 TI - Salivary levels of mutans streptococci associated with restorations: a case control study. AB - Our aim was to estimate whether restorative therapy with amalgam and composite resin could decrease salivary mutans streptococcal level, thus also decreasing the risk for other caries development. We selected a case group of 93 children with detectable salivary mutans levels (i.e., at least 1x10(4) cfu/ml), and a control group (n=93 subjects) with undetectable levels. Children had the same age (12 years), no extracted teeth, crowns, temporary fillings, and restorations other than amalgam and composite resin, and the two groups had similar gender distribution. We clinically examined children and recorded active caries, restorations and oral hygiene level by means of gingival bleeding on probing; we also investigated sucrose intake at breakfast. The case group had statistically significant higher prevalence of restorations (36.6% vs. 18.3%), active caries (44.1% vs. 12.9%), and bad oral hygiene (84.9% vs. 68.8%) than the control group. However, the logistic regression analysis showed that presence of active caries was the only significant variable associated with mutans streptococci (OR=4.0; p=0.0002), while the effects of sucrose intake and of restorations were marginally significant. This apparent contrast between statistical analyses was due to the concomitant presence, in children with detectable mutans streptococci level, of restorations and decayed teeth at the same time, and, on the basis of the multivariate analysis, presence of mutans streptococci in these children was explained by the presence of active caries, more than restorations. PMID- 11497087 TI - Genes of Mycobacterium tuberculosis H37Rv downregulated in the attenuated strain H37Ra are restricted to M. tuberculosis complex species. AB - By comparing gene expression of virulent Mycobacterium tuberculosis H37Rv and attenuated strain H37Ra, we previously detected six genes that appear to be markedly downregulated in the attenuated strain compared with the virulent one. Three of these genes, i.e. Rv1345, Rv2770c, and Rv0288, code for proteins that can be predictively associated to immunological or pathogenetic aspects of M. tuberculosis infection; the other genes, i.e. Rv2336, Rv1320c, and Rv2819c, code for proteins with unknown functions (Rindi et al., 1999). In this paper we searched for the above mentioned genes in Pvu II-digested genomic DNA of a number of mycobacterial species by southern blot analysis employing PCR-generated probes in high-stringency conditions. Hybridization signals were only found in species belonging to the M. tuberculosis complex, i.e., M. tuberculosis, M. bovis, including the BCG strain, and M. microti, but not in other mycobacterial species, including M. avium, M. intracellulare, M. malmoense, M. xenopi, M. kansasii, M. simiae, M. marinum, M. scrofulaceum, M. gordonae, M. fortuitum, and M. smegmantis. These results indicate that genes Rv1345, Rv2770c, Rv0288, Rv2336, Rv1320c, and Rv2819c are associated with the most virulent mycobacteria and further support their potential role in M. tuberculosis virulence. PMID- 11497088 TI - Solitary keratoacanthomas in immunocompetent patients: no detection of papillomavirus DNA by polymerase chain reaction. AB - The aetiology of keratoacanthoma (Ka) is unknown, but human papillomavirus (HPV) has been implicated in the pathogenesis of this lesion. To evaluate the role of HPV in the development of KA in the general population, 20 tissue specimens were analysed by polymerase chain reaction (PCR). To include a broad range of both cutaneous and mucosal HPV types, PCR was performed with two sets of degenerate primers. No HPV-DNA sequences were detected in any lesions analysed. These results do not support the hypothesis that HPV is involved in the etiology and pathogenesis of the KA in immunocompetent patients. PMID- 11497089 TI - Antibody response of dogs inoculated with a large number of cultured canine monocytes adsorbed with a calicivirus in advance. AB - Five conventional Beagle dogs were intravenously injected with ten million canine monocyte cells (Cn/K99) cultured in vitro (Kadoi, 2000) adsorbed with a strain of calicivirus originally isolated from lions (Kadoi et al., 1997). Another two Beagle dogs were injected similarly with the virus suspension solely as control. Serum samples were collected from these dogs at intervals and specific seroneutralizing antibody production against the virus was measured in vitro. A significantly higher antibody production was demonstrated in the five dogs group. A clear booster effect was also proved in the sera of the dogs after the second virus inoculation made on day 100. A possibility of antigen presentation function of non-self monocytes is suggested. PMID- 11497090 TI - Metabolic complications of HIV-1 antiretroviral therapy: the lipodystrophy syndrome. AB - The lipodystrophy syndrome is one of the complications reported with increased frequency in patients with HIV-1 infection receiving antiretroviral therapy. The wide range of prevalence estimates may be due to differing definitions, methods and patient populations. We described the various pathogenic theories and the morphological and metabolic alterations associated with this syndrome. Even if no effective treatment exists, a correct lifestyle, adequate diet and physical exercise seem to be very important. Moreover drug therapies should be used with care to avoid potentially harmful interactions with antiretroviral agents. Ideally, the future effort to define the mechanism of lipodystrophy would be multidisciplinary and would involve not only experts in AIDS research but also nutritionists, endocrinologists and cardiologists. PMID- 11497091 TI - Evaluation of the traumatic coma data bank computed tomography classification for severe head injury. AB - This study determines the interrater and intrarater reliability of the Traumatic Coma Data Bank (TCDB) computed tomography (CT) scan classification for severe head injury. This classification grades the severity of the injury as follows: I = normal, II = diffuse injury, III = diffuse injury with swelling, IV = diffuse injury with shift, V = mass lesion surgically evacuated, or VI = mass lesion not operated. Patients with severe closed head injury were included. Outcome was assessed using the Glasgow Outcome Score (GOS) at 3 and 6 months. Four observers, two of them classifying the scans twice, independently evaluated CT scans. Of the initial CT scans of 63 patients (36 males, 27 females; age, 34+/-24 years), 6.3% were class I, 26.9% class II, 28.6% class III, 6.3% class IV, 22.2% were class V, and 9.6% class VI. The overall interrater and intrarater reliability was 0.80 and 0.85, respectively. Separate analyses resulted in higher inter- and intrarater reliabilities for the mass lesion categories (V and VI), 0.94 and 0.91, respectively, than the diffuse categories (I-IV) 0.71 and 0.67. Merging category III with IV, and V with VI resulted in inter- and intrarater reliabilities of 0.93 and 0.78, respectively. Glasgow outcome scores after 6 months were as follows: 19 dead (30%), one vegetative (2%), five severely disabled (8%), 17 moderately disabled (27%), and 21 good recovery (33%). Association measures (Sommers' D) between CT and GOS scores were statistically significant for all observers. This study shows a high intra- and interobserver agreement in the assessment of CT scan abnormalities and confirms the predictive power on outcome when the TCDB classification is used. PMID- 11497092 TI - Defining acute mild head injury in adults: a proposal based on prognostic factors, diagnosis, and management. AB - The lack of a common, widely acceptable criterion for the definition of trivial, minor, or mild head injury has led to confusion and difficulty in comparing findings in published series. This review proposes that acute head-injured patients previously described as minor, mild, or trivial are defined as "mild head injury," and that further groups are recognized and classified as "low-risk mild head injury," "medium risk mild head injury," or "high-risk mild head injury." Low-risk mild injury patients are those with a Glasgow Coma Score (GCS) of 15 and without a history of loss of consciousness, amnesia, vomiting, or diffuse headache. The risk of intracranial hematoma requiring surgical evacuation is definitively less than 0.1:100. These patients can be sent home with written recommendations. Medium risk mild injury patients have a GCS of 15 and one or more of the following symptoms: loss of consciousness, amnesia, vomiting, or diffuse headache. The risk of intracranial hematoma requiring surgical evacuation is in the range of 1-3:100. Where there is one computed tomography (CT) scanner available in an area for 100,000 people or less, a CT scan should be obtained for such patients. If CT scanning is not so readily available, adults should have a skull x-ray and, if this shows a fracture, should be moved to the "high-risk" category and undergo CT scanning. High-risk mild head injury patients are those with an admission GCS of 14 or 15, with a skull fracture and/or neurological deficits. The risk of intracranial hematoma requiring surgical evacuation is in the range 6-10:100. If a CT scan is available for 500,000 people or less, this examination must be obtained. Patients with one of the following risk factors- coagulopathy, drug or alcohol consumption, previous neurosurgical procedures, pretrauma epilepsy, or age over 60 years--are included in the high-risk group independent of the clinical presentation. PMID- 11497093 TI - Delayed hemispheric neuronal loss in severely head-injured patients. AB - Recent experimental studies have revealed that traumatic brain injury as well as ischemic brain injury can cause chronic progressive neuronal damage. In the present study, we demonstrate previously unreported delayed cerebral atrophy on computerized tomography (CT) scans in severely head-injured patients. Seventeen severely head-injured patients who required mild hypothermia to control intracranial hypertension after the failure of conventional therapies were retrospectively analyzed. All 17 patients survived more than 1 year. Delayed neuronal loss (DNL) was observed in only eight of the 17 patients. Eight patients with DNL required longer durations of mild hypothermia to control intracranial hypertension than nine patients without DNL. Six of these eight patients with DNL achieved functional recovery despite progressive atrophic changes demonstrated on CT scans. On CT scans, DNL was characterized by (1) the sudden appearance at several months postinjury of a low-density area in the hemisphere ipsilateral to the injury; (2) the preservation of essential cortical structure although related white matter structures showed severe atrophic changes; and (3) no spread of the low-density area to the contiguous territory of the other main cerebral artery. It is concluded that focal primary injury to underlying brain, if severe enough, can result in delayed hemispheric atrophy. PMID- 11497094 TI - Detection of single- and double-strand DNA breaks after traumatic brain injury in rats: comparison of in situ labeling techniques using DNA polymerase I, the Klenow fragment of DNA polymerase I, and terminal deoxynucleotidyl transferase. AB - DNA damage is a common sequela of traumatic brain injury (TBI). Available techniques for the in situ identification of DNA damage include DNA polymerase I mediated biotin-dATP nick-translation (PANT), the Klenow fragment of DNA polymerase I-mediated biotin-dATP nick-end labeling (Klenow), and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). While TUNEL has been widely utilized to detect primarily double-strand DNA breaks, the use of PANT to detect primarily single-strand DNA breaks and Klenow to detect both single- and double-strand DNA breaks has not been reported after TBI. Accordingly, coronal brain sections from naive rats and rats at 0, 0.5, 1, 2, 6, 24, and 72 h (n = 3-5/group) after controlled cortical impact with imposed secondary insult were processed using the PANT, Klenow, and TUNEL methods. Cells with DNA breaks were detected by PANT in the ipsilateral hemisphere as early as 0.5 h after injury and were maximal at 6 h (cortex = 66.3+/-15.8, dentate gyrus 58.6+/-12.8, CA1 = 15.8+/-5.9, CA3 = 12.8+/-4.2 cells/x 400 field, mean +/- SEM, all p < 0.05 versus naive). Cells with DNA breaks were detected by Klenow as early as 30 min and were maximal at 24 h (cortex = 56.3+/-14.3, dentate gyrus 78.0+/-16.7, CA1 = 25.8+/-4.7, CA3 = 29.3+/-15.1 cells/x 400 field, all p < 0.05 versus naive). Cells with DNA breaks were not detected by TUNEL until 2 h and were maximal at 24 h (cortex = 47.7+/-21.4, dentate gyrus 63.0+/-11.9, CA1 = 5.6+/-5.4, CA3 = 6.9+/-3.7 cells/x 400 field, cortex and dentate gyrus p < 0.05 versus naive). Dual-label immunofluorescence revealed that PANT-positive cells were predominately neurons. These data demonstrate that TBI results in extensive DNA damage, which includes both single- and double-strand breaks in injured cortex and hippocampus. The presence of multiple types of DNA breaks implicate several pathways in the evolution of DNA damage after TBI. PMID- 11497095 TI - Potentiated endothelium-derived hyperpolarizing factor-mediated dilations in cerebral arteries following mild head injury. AB - Evidence in the literature suggests that endothelium-derived hyperpolarizing factor (EDHF) may act in a compensatory manner such that during conditions of compromised nitric oxide (NO), EDHF serves as a back-up mechanism. Given that constitutive NO synthase is chronically downregulated after head trauma, we tested the hypothesis that EDHF is potentiated following injury. Male adult rats were subjected to either sham injury (n = 27) or mild controlled cortical impact (CCI) injury (n = 26). Branches of the middle cerebral artery (MCA) directly within the contusion site were harvested either 1 or 24 h later, pressurized to 60 mm Hg in a vessel chamber and allowed to develop spontaneous tone. Relaxation to luminal application of adenosine triphosphate (ATP) was similar in all groups. Relaxation to ATP in the presence of L-NAME (N(G)-nitro-L-arginine methyl ester) and indomethacin was similar in all groups except for vessels isolated at 24 h following mild CCI injury. In this case, L-NAME and indomethacin had no effect on the ATP-mediated dilation. The ATP-mediated dilation in L-NAME and indomethacin treated MCA branches was inhibited by charybdotoxin, an inhibitor of large conductance Ca2+-sensitive K+ channels. These findings suggest that there is a significant potentiation of the EDHF-mediated dilation to ATP in cerebral arteries isolated at 24 h following mild CCI injury. PMID- 11497096 TI - Death of rat sympathetic ganglion cells in vitro caused by neurite transection: effect of extracellular calcium. AB - Calcium entry into neurons secondary to excitotoxic insults is believed to cause neuronal death after trauma and ischemia, but the role of calcium influx in neuronal death after neurite transection independent of excitotoxicity has not been clearly defined. This study assesses the effect of variations in extracellular calcium concentration ([Ca2+]e) from 50 nM to 5 mM on cell death, in 14-day-old cultures of dissociated sympathetic neurons from the superior cervical ganglia of newborn rats. The neurites were transected with a custom-made injury device, and cell death was assessed with propidium iodide and fluorescence microscopy. We found that neurite transection caused a significant increase (p < 0.05) in cell death at all [Ca2+]e studies, but there was no significant difference in mortality at the various [Ca2+]e. Cell death significantly increased between 2 and 24 h postinjury at all three [Ca2+]e. Cell death increased with decreasing distance between the cell body and the transection site, and there was a significant decrease in mortality at distances greater than 0.66 mm, irrespective of the [Ca2+]e. These results suggest that influx of extracellular calcium is not responsible for posttransection cell death, suggesting that calcium release from internal stores or calcium-independent cell death mechanisms are triggered by neurite transection. PMID- 11497097 TI - Multiparametric monitoring of brain under elevated intracranial pressure in a rat model. AB - Intracranial hypertension may develop in most patients exposed to traumatic head injury. In many cases, patients enduring elevated intracranial pressure (ICP) will incur morbidity or mortality. Several methods are used in animal models to investigate the influence of ICP elevation on physiological parameters. In this study, we developed a cisterna magna model by adding a mechanism for warming the mock cerebrospinal fluid (CSF) entering the cisterna space to a temperature of 37 degrees C and combined this method for ICP elevation with the multiparametric monitoring system (Multiprobe Assembly [MPA]). Using the MPA, we monitored, for the first time, mitochondrial NADH redox state as well as ionic homeostasis under elevated ICP in a rat model. In addition, we monitored cerebral blood flow (CBF) by laser Doppler flowmetry, ECoG (bipolar electrodes), and surface temperature. Blood pressure was measured in the cannulated femoral artery. The ICP (monitored by Camino probe) was elevated to 50-60 mm Hg for 13-15 min, followed by 2 h of recovery. The results show that CBF was decreased by 90%, while NADH was elevated by 80% as compared to the normoxic levels. Complete depolarization occurred as evidence by the decrease in extracellular Ca2+ and a significant increase in K+. All parameters recovered 10 min after reopening the cannula to the cisterna magna to air pressure. We conclude that ICP elevation through the cisterna magna infusion method, used simultaneously with multiparametric monitoring, supplies reliable information on the brain tissue metabolic state with intracranial hypertension in a rat model. PMID- 11497098 TI - Telemetric blood pressure monitoring in conscious rats before and after compression injury of spinal cord. AB - Abnormal cardiovascular control after spinal cord injury (SCI) results in hypotension soon after injury. Later, paroxysmal hypertension and bradycardia in response to sensory stimulation below the level of injury develop in most people with SCI. In this study, we used a radiotelemetry system, in rats (n = 7), to investigate the effect of a clinically relevant compression model of SCI at T5 spinal segment on mean arterial pressure (MAP) and heart rate (HR) at rest and in response to colorectal distension. The transducers were implanted 1 month before clip compression (50-g) injury and continuous recording of MAP and HR was established for a period of 2.5 months. SCI was associated with hypotension (86+/ 3 mm Hg) at 1 day after injury. In the following 2 days, MAP gradually returned to preinjury levels. By contrast, HR increased at 1 day after SCI and remained unchanged thereafter. Three days after SCI, colorectal distension caused an increase in MAP of 8+/-2 mm Hg accompanied by bradycardia (-18 bpm). One week after SCI, colorectal distension induced an increase in MAP of 9+/-2 mm Hg and bradycardia (-41 bpm). In the following days, the magnitude of reflex hypertension gradually increased, reaching 21+/-4 mm Hg at 1.5 months after SCI. In summary, our data show that resting MAP rapidly returns to control values after SCI. Episodic hypertension associated with autonomic dysreflexia can develop in rats within 1 month after incomplete SCI. PMID- 11497099 TI - A lateral mass fracture of C1 associated with left vertebral artery and mid basilar artery occlusion. AB - Extreme acceleration and deceleration forces as well as axial loading are exerted at the occipito-cervical junction of drivers involved in high-velocity motor vehicle accidents, especially with fastened seatbelts. Injury at this level, usually lethal, can go unrecognized despite modern emergency management of the unconscious patient. A precise neurologic and radiographic workup of damage to this area is often not possible or overlooked in the initial phase of such severe trauma. We describe a patient with multiple injuries who sustained a left vertebral artery occlusion associated with a left-sided lateral mass fracture of C1 and a basilar artery occlusion resulting in a locked-in syndrome after an automobile accident. PMID- 11497100 TI - The role of sphingosine and its derivatives in the regulation of Ca2+ homeostasis. AB - The role of sphingosine and sphingosine 1-phosphate in the regulation of Ca2+ homeostasis is reviewed. Available data suggest that there are at least three main pathways by which sphingosine and its metabolites affect Ca2+ fluxes in different cell types: (1) indirect action on Ca2+ stores, mediated by other transduction pathways; (2) direct action on the receptors of Ca2+ channels and Ca2+ pumps which are localized at the membranes of Ca2+ stores; (3) indirect action mediated by the regulation of expression of the channel-forming protein Bcl-2, which incorporates into membranes of Ca2+ stores. The effects of sphingosine and its metabolites on Ca2+ homeostasis via mechanisms (1)-(3) are considered. The combination of the mechanisms by which sphingosine affects Ca2+ signalling pathways is tissue-specific. Sphingosine and its metabolites can be physiological modulators of the intracellular Ca2+ signalling pathways. PMID- 11497101 TI - UV-Induced destruction of light-harvesting complexes from purple bacterium Chromatium minutissimum. AB - We studied UV-induced photodestruction of the native forms of bacteriochlorophyll a (Bchl a) from chromatophores and light harvesting complexes (LHC) of the sulphur photosynthetic bacterium Chromatium minutissimum. Irradiation of chromato phores with 365-nm light (Soret band) or 280-nm light (absorption region of aromatic amino acids) led to the destruction of all long-wavelength forms of Bchl a. The quantum yields of photodestruction produced by the 280-nm light was higher than that produced by the 365-nm light. For the spectral forms of Bchl a absorbing at 850 nm and 890 nm, the difference was about one order of magnitude, and for the form absorbing at 800 nm the difference was almost two orders of magnitude. Similar UV sensitivity was observed for the Bchl a forms from isolated LHC. As a rule, the quantum yields of photodestruction induced by UV irradiation at 280 nm were about 100-1000 times higher (approximately 10(-3)-10(-4)) than that upon red light irradiation (approximately 10(-6)-10(-7)). We found that irradiation of chromatophores at 280 nm resulted in a crosslink between the core and peripheral LHC. PMID- 11497102 TI - The outward component of photoinduced current in chloroplasts of Peperomia metallica. AB - The photoinduced currents in whole chloroplasts of Peperomia metallica were studied using suction electrodes and single-turnover flashes. The kinetic profile of the photocurrent contained a minor outward component (rise time, 100 micros). Local application (from the inside of the pipette) of a photosystem 2 inhibitor, DCMU, rapidly suppressed the outward current; conversely, addition of DCMU to the outer medium produced a transient stimulation of the outward component. Permeabilization of the tip-located membrane fragments with Triton X-100 eliminated the outward current, but had no significant influence on the inward current. The data suggest that the outward current originated in the tip-located nonruptured portions of the thylakoid membrane. Different involvement of two photosystems in the generation of the outward current indicates that granal thylakoids enriched with photosystem 2 are less susceptible to the rupture in the pipette tip as compared with stromal thylakoids. PMID- 11497103 TI - Potassium gradients in the growing hyphae of Neurospora crassa. AB - X-ray electron probe microanalysis of a Neurospora crassa hyphal preparation revealed a significant heterogeneity of the potassium content in the hyphae. Low potassium areas were found in a close proximity of the growing tip and in the distal part of the hyphae in the submembrane area of about 1 microm width. The intracellular potassium concentration was shown to change more than 3-5 times from apical to distal regions of the leading hyphae, the potassium contents in the apical part of the hyphae usually being minimal. We suggest that the content of intracellular potassium is determined by the volume occupied by free intracellular water, i.e., by the size of the space available for diffusion of substances, including inorganic ions. PMID- 11497104 TI - Relation between ionic and hormonal regulation of proton pumps in red beet tonoplast. AB - Effects of indoleacetic, abscisic, gibberellic acids and kinetin on the hydrolytic activity of the proton pumps H+-ATPase (EC 3.6.1.3) and H+ pyrophosphatase (EC 3.6.1.1) under non-optimal ionic conditions were studied in tonoplasts isolated from stored roots of red beet (Beta vulgaris L. cv. Bordo). We showed that ionic composition markedly affected the hormonal regulation. H+ pyrophosphatase was more sensitive to the ionic and hormonal regulation than H+ ATPase. Magnesium ions are essential for the maximal manifestation of hormonal stimulation. PMID- 11497105 TI - Antigenic relationship and functional properties of Yersinia porins. AB - We have studied the molecular structure and functional properties of major pore forming proteins isolated as peptidoglycan (PG)-protein complexes from four Yersinia species (Y. intermedia, Y. enterocolitica, Y. kristensenii and Y. frederiksenii) cultured as various temperatures. Despite the close antigenic relationship, Yersinia porins revealed different functional properties. When reconstituted in model membranes, the PG-protein complexes induced conductance which was different for the "cold" (grown at 6-8 degrees C) and "warm" (grown at 37 degrees C) variants of microbial cultures. We conclude that the functional state of Yersinia porins in the outer membrane depends on the cultivation temperature. PMID- 11497106 TI - Steroid-induced conformational changes of FITC-labelled sarcoplasmic reticulum Ca2+-ATPase. AB - Interactions between transmembrane and cytoplasmic domains of Ca2+-ATPase from sarcoplasmic reticulum (SR) have been studied. To affect the hydrophobic transmembrane domain, we used four amphiphilic steroids - esters of a dibasic acid and 20-oxypregnene. All four steroids contained cholesterol-like nuclei and differed by the structure of side chains. Steroids with carboxyl groups in the side chains inhibited the rates of ATP hydrolysis and Ca2+ transport, whereas a steroid without the carboxyl group did not appreciably affect Ca2+-ATPase function. Fluorimetric titration of FITC-labelled Ca2+-ATPase in SR vesicles by Nd3+ showed that steroids increased the apparent dissociation constant for Nd3+ bound to the hydrolytic site, the potency order of the steroids being the same as for the sterol-induced inhibition of the hydrolytic activity of Ca2+-ATPase. These results suggest structural changes in the active site. Ca2+ transport was inhibited more efficiently by steroids than the hydrolytic activity of the enzyme. This could be partially due to the increase of the membrane passive permeability induced by steroids, which, in turn, reflected the efficiency of the interaction of the steroids with lipid bilayers. The effects of the steroids were largely dependent on their amphiphilicity (the availability of polar groups in regions A and D), the structure of the side chains, and, possibly, on the distance between the molecular polar groups. We suggest that the inhibition of hydrolytic and transport functions of Ca2+-ATPase in the SR membrane is due to the interaction of the steroids with the transmembrane alpha-helical segments. PMID- 11497107 TI - PH-dependence of detergent-induced hemolysis and vesiculation of erythrocytes. AB - The influence of pH of the medium on the parameters of detergent-induced fast hemolysis and vesiculation of human erythrocytes was studied. In the range of pH 6.3-7.2 neither the extent nor the rate of the vesiculation induced by 25 microM sodium dodecyl sulfate (SDS) changed. However, a decrease of pH from 8.0 to 5.8 strongly modified both the extent and the rate of the hemolysis induced by SDS. Within the range of pH 8.0-6.4, the effect can be ascribed to the increase of the positive charge of the membrane. This could lead to the accumulation of the membrane-bound anion detergent and, hence, to the change of the hemolysis parameters. Non-charged detergent Triton X-100 did not display any pH-dependence. At pH between 6.4 and 5.8 the extent and rate of hemolysis changed in a complicated manner. The kinetic curves of hemolysis could be approximated by a single exponential within the pH range between 8.0 and 7.2. Upon further reduction of pH, a second exponential component, with a larger time constant, appeared in the kinetic curves. At 5.8 < pH < 7.2, the contribution of the "fast" hemolysis dropped virtually to zero, with pK about 6.0. This points to a structural transition of the membrane, possibly involving histidine. We suggest that the parameters of the detergent-induced hemolysis are sensitive to the changes of the charge and structural state of erythrocyte membrane. PMID- 11497108 TI - Dielectro-deformations of erythrocyte: analysis of the ellipsoidal shear model. AB - Using the theoretical analysis within the framework of the proposed ellipsoidal shear electromechanical model of erythrocyte, the main mechanisms and relationships have been established and studied for the deformations of erythrocytes caused by a spatially homogeneous high-frequency electric field. The main types of the stress-strain curves characteristic of stationary and dynamic deformations caused by the rectangular-pulse and harmonic modulations of the field amplitude have been calculated. The relationship has been established between the parameters of essentially nonlinear stress-strain curves and mechanical, electric, and geometric parameters of erythrocyte. The impossibility of unlimited elongation of erythrocyte by the field, due to the conservation of the cell volume and surface area, has been shown, and the dependence of the maximum possible elongation of the cell on its volume has been calculated. It has been shown that the relaxation time of dynamic deformations of erythrocyte in the presence of an electric field considerably differs from that characteristic of the membrane material and sharply decreases with the increase of the initial elongation of the cell. PMID- 11497109 TI - Redox properties of local anesthetics: A structural determinant of closed channel blockers in BTX-modified Na+ channels. AB - Single channel analyses and macroscopic current measurements have shown that benzocaine is a predominantly closed channel blocker in BTX-modified Na+ channels; cocaine is an open channel blocker; and tetracaine, a dual channel blocker (Wang & Wang, 1994; Wang et al., 1994). The reason for such a selective state-dependent block by local anesthetics in BTX-modified Na+ channels is not clear. We assessed the redox properties of tetracaine, benzocaine, cocaine, and various derivatives by their ability to donate electrons to radical intermediates of eosin dye excited by visible light. Electron-donor properties of the drugs were previously proposed to be involved in Na+ channel blockade (Marinov, 1991). Our results provide evidence that redox properties of tetracaine, benzocaine, and their homologs correlate with their ability to enhance Na+ channel inactivation in BTX-modified Na+ channels. This correlation may be explained in terms of the previously proposed redox model of ion channels. PMID- 11497110 TI - Using qualitative methods to elicit young people's perspectives on their environments: some ideas for community health initiatives. AB - This paper describes qualitative methods used in a research project for the former Health Education Authority, exploring Putnam's concept of 'social capital' in relation to children and young people's well-being and health. Putnam's conceptualization of social capital consists of the following features: trust, reciprocal support, civic engagement, community identity and social networks, and the premise is that levels of social capital in a community have an important effect on people's well-being. Research was carried out with 102 children aged between 12 and 15 in two relatively deprived parts of a town in southeast England. The paper describes the research setting, methods, consent process and ethical issues that arose. It explores how the methods generated different forms of interconnected data, giving rise to a number of health/well-being-related themes. The paper concludes that using a range of methods, including visual methods, has helped to explore quality of life issues for children that are usually neglected in studies of young people's health-related behaviours. PMID- 11497111 TI - Knowledge, attitudes and self-care practices related to sun protection among secondary students in Australia. AB - The aim of this research was to ascertain changes in sun-related knowledge, attitudes and self-care practices among Australian secondary school students between 1993 and 1996. Two cross-sectional surveys of sun-related attitudes, beliefs and behavior of young people aged 12-17 years of age, were conducted in 1993 and 1996. Over 80% of adolescents at both time periods knew about the issues related to skin cancer prevention, frequency of burning and burning on cloudy days. Adolescent attitudes had shifted positively in the areas of staying inside in 1996 [relative risk (RR): 1.13; 95% confidence interval (CI): 1.09-1.17] and staying under shade in 1996 (RR: 1.16; 95% CI: 1.13-1.18). Desire for a moderate or dark tan was lower in 1996 (45%) than in 1993 (50%). Respondents reported that they were less likely to wear brief clothing to get a suntan in 1996 (RR: 0.81; 95% CI: 0.78-0.84) and were significantly more likely to stay in the shade in 1996 (RR: 1.19; 95% CI: 1.16-1.23). We conclude that there has been a shift in attitudes towards use of shade and avoidance of unnecessary exposure, and away from use of sunscreens and sunglasses. The results suggest that adolescents may be more ready to accept structural changes that move desired activities out of the sun. PMID- 11497112 TI - Young people in 'drinking' societies? Norwegian, Scottish and Swedish adolescents' perceptions of alcohol use. AB - The paper studies young people's reported drinking behaviors and their views on various social aspects of alcohol, utilizing a sample of over 4000 rural adolescents aged 11.8-16.5 years in Norway, Scotland and Sweden. The methodology employed includes a common questionnaire and a range of varying qualitative approaches (essays and focus group interviews). The various venues and drinking contexts used by young people, their motives for drinking, and their 'learning' experiences with alcohol are described. Beyond nationality, the most powerful predictors of 'high' drinking are 'involvement with friends' and 'participation in commercial leisure'. The predictors for 'low' drinking are 'involvement in activities with parents' and 'parental concerns about drinking'. Results show that Scottish teenagers drink most, Norwegians least and no differences in the predictor variables are found that can explain this. Results are discussed in relation to social and cultural differences, and illustrated by quotations from rural young people in Scotland and Sweden. PMID- 11497113 TI - Predicting condom use with casual and steady sex partners among drug users. AB - In many Western societies, drug users are accountable for heterosexual transmission of HIV. Although various studies have shown that safe sex is not a common practice among drug users, we still have little understanding of the factors that determine their use of condoms in their sexual relationships with steady or casual partners. The objective of this study is to gain more insight into the determinants of drug users' condom use with steady and casual sex partners. In a study conducted among 103 Dutch drug users, the participants indicated their intentions, vulnerability, attitude, personal norms, subjective norms, descriptive norms, self-efficacy and response efficacy regarding condom use with steady and casual partners. The results showed that some drug users had borrowed used syringes and that most drug users practise unsafe sex, especially with steady partners. Most drug users were hardly motivated to start using condoms. The study further showed that antecedents of condom use differ with regard to steady sex partners and casual sex partners. Self-efficacy and subjective norms were predictors of intentions to use condoms with steady partners; intention to use condoms with casual partners was only predicted by self-efficacy. These differences should be taken into account when designing AIDS prevention interventions to encourage use of condoms. PMID- 11497114 TI - Evoked fear and effects of appeals on attitudes to performing breast self examination: an information-processing perspective. AB - The effect of fear arousal on attitude toward participating in early detection activities [i.e. breast self-examination (BSE)] was studied from an information processing perspective. It was hypothesized that fear arousal motivates respondents to more argument-based processing of fear-relevant persuasive information. Respondents first read information about breast cancer in which fear was manipulated. After measuring fear arousal, respondents read a persuasive message about performing BSE. Analyses with reported fear, but not manipulated fear, found support for the hypothesis. Respondents who reported mild fear of breast cancer based their attitude toward BSE more on the arguments provided than respondents who reported low fear of breast cancer. This finding suggests that the use of fear arousal may be an efficient tool in health education practice. However, alternative interpretations are provided, in addition to the suggestion to be careful with using fear arousal in health education messages. PMID- 11497115 TI - Occupational health and safety amongst sex workers: a pilot peer education resource. AB - This paper presents an account of a pilot project to design and implement an innovative, sex-worker-driven approach to peer education in London. A 28-min, double-sided tape cassette containing extracts of sex workers talking candidly about their work was compiled from oral history recordings. The aims were to pilot process issues in compiling the collaborative resource, and to stimulate discussion of health and safety issues, exploring whether the resource could potentially enhance positive changes in sex workers' knowledge and awareness, self-esteem, and lifeskills. In a limited pilot distribution, 15 sex workers and seven outreach workers in London listened to the tape and completed short evaluation questionnaires. The paper sets out strengths and weaknesses of the pilot method, linking these to broader critical reflections on issues raised about peer education to, for and by sex workers. PMID- 11497116 TI - A comparison of adolescent moderate-to-vigorous physical activity participation in relation to a sustained or accumulated criterion. AB - Contemporary guidelines for young people advocate both a sustained and accumulative approach to moderate-to-vigorous physical activity (MVPA) participation. In order to investigate the behavioural significance of applying these approaches, this study assessed if differences in adolescent MVPA occurred when either a sustained or accumulated criterion was adopted. Using heart rate thresholds indicative of intensity, the physical activity of 25 adolescents was assessed by monitoring heart rate over 3 days. Results indicated that differences in MVPA participation did exist when different approaches were used, in that the majority of adolescents were active with respect to an accumulated criterion yet inactive with respect to a sustained criterion. Such a disparity may be due to accumulative MVPA being more characteristic of young people's natural activity behaviour. Such findings have strong implications for practitioners seeking to counter young people's inactivity, in that the application of an accumulative approach may be more effective at initiating and establishing an habitual activity behaviour than that of a sustained approach. PMID- 11497117 TI - Mass education for obesity prevention: the penetration of the BBC's 'Fighting Fat, Fighting Fit' campaign. AB - The study aimed to evaluate the BBC's 'Fighting Fat, Fighting Fit' campaign's success in achieving public awareness of the need for obesity prevention, and putting over its message of healthy eating and increased physical activity. Demographic factors associated with awareness of the campaign were assessed. Data were collected from a national representative British sample as part of the ONS Omnibus survey in March 1999. Questions included weight and height, recognition of the campaign name, recall of the content of the campaign, and participation in the campaign registration scheme. More than half of the respondents had heard of the campaign and 30% recalled the healthy lifestyle messages, although fewer than 1% registered to participate in the scheme. Awareness of the campaign was high in all socio-economic groups, but memory for the healthy lifestyle message was significantly poorer in those with lower levels of education and from ethnic minority groups. Disappointingly, awareness was also no higher in overweight than normal weight respondents. The results strongly support the effectiveness of the campaign in publicizing the issue of increasing prevalence of obesity and the need for lifestyle change, but suggest that different approaches might be needed to maximize participation from groups most in need of lifestyle change. PMID- 11497118 TI - Using the mass-media to target obesity: an analysis of the characteristics and reported behaviour change of participants in the BBC's 'Fighting Fat, Fighting Fit' campaign. AB - The study aimed to assess the characteristics and reported behaviour change of participants in the BBC's 'Fighting Fat, Fighting Fit' (FFFF) campaign. A postal questionnaire survey was sent to a random sample of 6000 adults registering with the FFFF campaign at the start of the campaign and 5 months later. Demographic characteristics, weight, eating behaviour and activity patterns were assessed. In total, 3661 respondents completed the baseline questionnaire and 2112 (58%) of these completed a follow-up evaluation questionnaire 5 months later. The majority of evaluation participants were women and classified as 'overweight' or 'obese'. Participants reported significant reductions in weight, and in fat and snack intake, and significant increases in exercise levels, and in fruit, vegetable and starch intake during the 6 months of the campaign. These effects remained significant if non-responders were assumed to have made no change. These results show that mass-media campaigns might make a contribution to weight control at the population level, but particular subgroups such as men and people under 25 may require specifically targeted campaigns. In addition, whilst such campaigns may initially attract obese people, they may be more likely to drop out of the campaign than overweight and normal weight individuals. PMID- 11497119 TI - Effect of culturally appropriate health education on glycaemic control and knowledge of diabetes in British Pakistani women with type 2 diabetes mellitus. AB - Type 2 diabetes is a growing problem in people of South Asian origin. It is associated with severe complications if it is not adequately controlled. This paper is a secondary assessment of 105 British Pakistani women within a larger randomized controlled trial of 200 Pakistani patients with diabetes. The trial used one-to-one structured diabetes health education, delivered by a linkworker with pictorial flashcards as a visual aid. Earlier published results from this study have shown that the women in the study knew less about diabetes and had poorer glycaemic control than men, which is why this assessment was performed to see what happened to them when they received appropriate health education. All patients were assessed before and 6 months after intervention by questionnaire and haemoglobin A1c blood tests to measure their overall blood sugar control. Nearly everyone improved their knowledge scores after 6 months in the intervention group, with women showing a catch-up improvement such that they equalled men. Multiple regression analysis found that glycaemic control improved in women receiving health education. Although this method of health education improved knowledge and glycaemic control in women in this sample, illiterate women did not do as well as their literate peers, continuing to score less on knowledge parameters. They also did not show an improvement in glycaemic control. Further work is needed to discover methods that will reach this sizeable subsection of the community. PMID- 11497120 TI - Tuberculosis (TB) and HIV linked, joint efforts needed. PMID- 11497121 TI - Tackling obesity in England. PMID- 11497122 TI - Single K+ channels in endocrine cells dispersed from the cricket (Gryllus bimaculatus) corpora allata. AB - Single channel currents were recorded from cell-attached patches of endocrine cells of the adult male cricket corpora allata. Three distinct types of K+ channels were identified; a weak inward rectifier (Type 1), a strong inward rectifier (Type 2) and a weak outward rectifier (Type 3). The type 1 channel had a slope conductance of 191 +/- 9 pS (n = 4) at negative membrane potentials (Vm) and 101 +/- 6 pS (n = 6) at positive Vm. In addition, the channel showed fast open-closed kinetics at negative Vm and slow open-closed kinetics at positive Vm. The open probability (Po) of this channel was strongly voltage-dependent at positive Vm, but less voltage-dependent at negative Vm. The reversal potential was not modified significantly by the substitution of gluconate for external Cl- but was modified after N-methyl-D-glucamine (NMDG+) was substituted for external K+, according to the Nernst equation for a K+-selective channel. The type 2 channel had a slope conductance of 44 +/- 2 pS (n = 5) at negative Vm, but no detectable outward current was observed at positive Vm. This channel showed very slow open-closed kinetics at negative Vm and its Po was not voltage-dependent. The type 3 channel had a limit conductance of 55 +/- 12 pS (n = 3) at negative Vm and 88 +/- 10 pS (n = 3) at positive Vm. This channel showed slow open-closed kinetics at negative Vm and fast open-closed kinetics at positive Vm. The Po for the channel was voltage-dependent at positive Vm but was voltage-independent at negative Vm. These three types of K+ channels may be important for the control of the resting membrane potential, and may thus participate in the regulation of Ca2+ influx and juvenile hormone secretion in corpora allata cells. PMID- 11497123 TI - The sleeper Bostrichthys sinensis (family Eleotridae) stores glutamine and reduces ammonia production during aerial exposure. AB - Bostrichthys sinensis inhabits brackish water, living in the crevices of the river mouths of Shang Xi and Guangdong, China. In its natural habitat, it may encounter aerial exposure frequently during low tides, and it usually remains quiescent in the absence of water. Upon aerial exposure in the laboratory, the ammonia excretion rate decreased to one-fourth that of the submerged control. Although all the enzymes of the ornithine-urea cycle were detected in the liver of this fish, the activity of hepatic carbamoyl phosphate synthetase was too low for the cycle to be functioning. Indeed, ammonia accumulated in the tissues and was not converted to urea. Results indicate that ammonia produced through amino acid catabolism was detoxified to glutamine during the first 24 h of aerial exposure. The excess amount of glutamine stored in the muscle during this period couldaccount approximately for the reduction in ammonia equivalent excreted. There was indeed a significant increase in the activity of glutamine synthetase from the liver of specimens exposed to terrestrial conditions. In contrast to the production of alanine, formation of glutamine is energetically expensive. Since B. sinensis remained relatively inactive on land, the reduction in energy demand for muscular activity might provide it with the opportunity to exploit glutamine formation as a means to detoxify ammonia. After 72 h of aerial exposure, B. sinensis reduced internal ammonia production, possibly through reductions in proteolysis and amino acid catabolism, to avoid excessive accumulation of ammonia. PMID- 11497125 TI - Inhibitory non-genomic effects of progesterone on Na+ absorption in epithelial cells from Xenopus kidney (A6). AB - The effect of the steroid hormone progesterone on transepithelial sodium transport was measured in confluent monolayers of the A6-cell line from Xenopus kidney. Apical permeabilization with Amphotericin B enabled us to measure the Na+/K+-pump current, and current-fluctuation analysis was used to analyze changes in apical channel density and gating characteristics. Basolateral progesterone (22.2 microM) had a rapid inhibitory effect on the Na+/K+-pump current, and a corresponding decrease in Na+ channel density. The effect occurred within some minutes and took about 50 min to reach a new steady state, in which 45% of the macroscopic current (ISC) was inhibited. Progesterone also inhibits the hypo osmotic stimulation of Na+ channels which occurs in untreated monolayers. Compared with the known effects of adrenal steroids, our results show a rapid inhibitory action of a steroid hormone on Na+ absorption. The time profile of the progesterone effect suggests, at least in the first minutes, a non-genomic action of progesterone. PMID- 11497124 TI - Regulation of cAMP-sensitive colonic epithelial Na+ channel in oocyte expression system. AB - In amphibian epithelia and in cortical collecting duct the antidiuretic peptide arginine-vasopressin (AVP) stimulates activity of epithelial Na+ channels (ENaCs). Generally, the AVP action upon Na+ (re)absorption is believed to be a cAMP/protein-kinase-A mediated mechanism. In the Xenopus oocyte expression system, however, a clear stimulation of ENaC activity by cAMP could not be reproduced with channel subunits cloned from A6 cells or rat colon. We have recently shown that membrane-permeant 8-(4-chlorophenylthio)-cAMP (cpt-cAMP) stimulates activity of a hybrid ENaC in Xenopus oocytes, that consists of an alpha-subunit cloned from guinea-pig colon and the beta- and gamma-subunit originating from rat colon (gpalpharbetagammaENaC). In the present study, we have further investigated the mechanisms by which cpt-cAMP upregulates gpalpharbetagammaENaC activity. Interestingly, we found AVP to stimulate the gpalpharbetagammaENaC in oocytes. Also, treatment with GTP-gamma-S largely activated this channel. In contrast, as a conflicting result, forskolin had no stimulatory effect on the cAMP-sensitive gpalpharbetagammaENaC. Experiments with Brefeldin A (BFA) or nocodazole suggested that only a minor part of cpt-cAMP induced activation is probably due to an additional translocation of channel proteins into the oocyte membrane. In conclusion, the stimulatory effect of synthetic cpt-cAMP does not seem to be exclusively provided by classical cAMP/PKA associated transduction mechanisms, i.e., as in A6 cells. PMID- 11497126 TI - Distinguishing two types of gray mullet, Mugil cephalus L. (Mugiliformes: Mugilidae), by using glucose-6-phosphate isomerase (GPI) allozymes with special reference to enzyme activities. AB - The resident and migratory types of gray mullet, Mugil cephalus, on the coast of Taiwan can not be separated morphologically. Allozyme analysis was applied to estimate genetic variation between the two types of gray mullet and to test whether they belong to different populations. After starch gel electrophoresis, different allelic frequency spectra of glucose-6-phosphate isomerase-A (GPI-A) between stocks was observed. The resident stock contained Gpi-A(135) and Gpi A(100), whereas the migratory type contained Gpi-A(100) only. In addition, GPI activities of locus A showed two distinct profiles between the two alleles. The results broadly revealed that Gpi-A allelic frequency was not regulated by temperature changes even after 6 months of thermal acclimation. This suggests that natural selection may play a role in shaping the allelic frequency change during the migratory journey. These findings suggest that the Gpi-A allelic difference can be used for population discrimination. PMID- 11497127 TI - Changes in heart rate are important for thermoregulation in the varanid lizard Varanus varius. AB - Laboratory studies and a single field study have shown that heart rate in some reptiles is faster during heating than during cooling at any given body temperature. This phenomenon, which has been shown to reflect changes in peripheral blood flow, is shown here to occur in the lizard Varanus varius (lace monitor) in the wild. On a typical clear day, lizards emerged from their shelters in the morning to warm in the sun. Following this, animals were active, moving until they again entered a shelter in the evening. During their period of activity, body temperature was 34-36 degrees C in all six study animals (4.0-5.6 kg), but the animals rarely shuttled between sun and shade exposure. Heart rate during the morning heating period was significantly faster than during the evening cooling period. However, the ratio of heating to cooling heart rate decreased with increasing body temperature, being close to 2 at body temperatures of 22-24 degrees C and decreasing to 1.2-1.3 at body temperatures of 34-36 degrees C. There was a significant decrease in thermal time constants with increasing heart rate during heating and cooling confirming that changes in heart rate are linked to rates of heat exchange. PMID- 11497128 TI - Free-ranging heart rate, body temperature and energy metabolism in eastern grey kangaroos (Macropus giganteus) and red kangaroos (Macropus rufus) in the arid regions of South East Australia. AB - Eastern grey kangaroos (Macropus giganteus) are generally regarded as mesic inhabitants. Even though access to drinking water in permanent stock watering troughs is commonly available, these animals are still found in only low densities in arid pastoral areas. We hypothesised that the differential success of red and grey kangaroos in the arid zone may be due to higher energy requirements of M. giganteus with a concomitant need for increased food, rather than limitations imposed by inadequate water access. We set out to test this by indirectly measuring energy expenditure through the monitoring of heart rate by radio telemetry in semi-free-ranging eastern grey and red kangaroos (Macropus rufus). Radio telemetry measurements of heart rate were calibrated against oxygen consumption and were used in the assessment of energy expenditure of animals maintained in an 8-ha enclosure in the arid zone of southeast Australia. Heart rate provided a reliable estimate of oxygen consumption. This well-correlated relationship was curvilinear and was established for each individual. Behavioural observations revealed that both kangaroo species spent most of the day in low energy demanding activities. M. rufus were more active at night whilst M. giganteus were more active in the early mornings and late afternoons. Like other marsupials, both species had low field metabolic rates (FMRs). However, M. giganteus in keeping with their mesic history had higher FMRs than the more arid adapted M. rufus, particularly during water restriction. Body temperature telemeters revealed a further species difference in that under hot conditions when water is freely available, M. rufus exhibits a higher and more labile daytime body temperature than M. giganteus. During the hottest part of the day M. giganteus maintain body temperature, relying upon increased evaporative cooling mechanisms, such as licking. Indeed, only when access to drinking water was restricted was thermolability evident in M. giganteus. Differences in behaviour and concomitant energy expenditure may thus contribute substantially to the divergent distribution and abundance of these two kangaroo species. PMID- 11497129 TI - Aldosterone upregulates purinergic responses in larval amphibian skin epithelium. AB - Bullfrog tadpoles respond to apical application of 100 microM amiloride, acetylcholine (ACh) or ATP with a sharp transient inward (apical to basolateral) cation current. In adult skin, amiloride blockable transepithelial Na+ transport is upregulated by the hormone aldosterone. Tadpoles were treated in vivo with aldosterone and changes in short circuit current (Isc) in response to apical application of ATP were determined. Bullfrog tadpoles were exposed to aldosterone (10(-6) M) for periods ranging from 3 h to 60 h. Skins from 60-h aldosterone treated animals showed a two- to three-fold increase in apical ATP-activated short circuit current when compared to animals treated with vehicle alone. Sodium replacement with a large, nonpermeable cation resulted in no measurable increase in Isc after exposure to ligand, consistent with ATP activation of an inward cation current and not chloride efflux. Activation/desensitization time courses and treatment with blockers revealed no measurable differences between aldosterone-treated and non-treated skins. Activation by amiloride and ACh gave essentially identical results. Studies with RT/PCR showed significant increases over controls of levels of mRNA associated with P2X channels. Given these data, our working hypothesis is that all three ligands activate the same process that exhibits both purinergic and cholinergic characteristics. These data are consistent with aldosterone upregulation of ATP gated channels expressed in the apical membrane of larval frog skin. PMID- 11497130 TI - Effect of salinity on flavin-containing monooxygenase expression and activity in rainbow trout (Oncorhynchus mykiss). AB - In order to obtain more information about the physiological role(s) of flavin containing monooxygenases (FMOs) in euryhaline teleost fishes, two experimental series were performed using adult and juvenile rainbow trout (Oncorhynchus mykiss). Cannulated adult trout were exposed to freshwater or 21% seawater for 48 h, whereas juvenile trout were acclimated to one of four different salinities: freshwater, 7%, 14%, or 21% during a 2-week period. FMO expression and activity were determined in red blood cells (RBC), liver, gill, kidney, gut, heart and brain. Furthermore, the content of trimethylamine oxide (TMAO; an FMO metabolite and an osmolyte) as well as urea were determined in various tissues. FMO expression and activity increased significantly and in a salinity dependent manner in osmoregulatory organs (gills, kidney and gut) in both juveniles and adult trout and, furthermore, in RBC in adults. No significant changes were observed in liver or heart. Urea content increased significantly and in a salinity dependent manner in all tissues, whereas TMAO was accumulated primarily in muscle tissue. Salinity dependent adjustment of FMO expression and activity primarily in osmoregulatory organs as well as regulation of TMAO content in muscle is consistent with previous studies showing an association of FMO with osmoregulation in euryhaline teleosts. However, the lack of a parallel increase of TMAO with urea in other tissues of fish at high salinity indicates other mechanisms of protection from intracellular urea may exist in non-muscular tissues. PMID- 11497131 TI - Temporal organisation of hibernation in wild-type and tau mutant Syrian hamsters. AB - The temporal pattern of hibernation was studied in three genotypes of Syrian hamsters with different circadian periodicity to assess a potential circadian control of alternating torpor and euthermy. We recorded the pattern of hibernation by measuring activity in continuous dim light and constant environmental temperature (6 +/- 1 degrees C). In spite of differences in the endogenous circadian period of three genotypes (tau +/+: approximately equals 24 h, tau +/-: approximately equals 22 h, and tau -/-: approximately equals 20 h) torpor bout duration was statistically indistinguishable (tau +/+: 86.9+/-5.3 h; tau +/-: 94.2+/-3.3 h; tau -/-: 88.8+/-6.2 h). The time between two consecutive arousals from torpor showed unimodal distributions not significantly different between genotypes. The first entry into torpor occurred within the active phase of the circadian cycle in all genotypes whereas the first arousal from torpor appeared to be timed randomly with respect to the prior circadian cycle. The amplitude of the activity rhythm was lower after hibernation compared with the amplitude before hibernation. The results suggest that in the Syrian hamster the circadian system does not control periodicity of torpor and arousal onsets in prolonged hibernation at 6 degrees C. PMID- 11497132 TI - Survival and quality of life 12 years after ICU. A comparison with the general Norwegian population. AB - OBJECTIVES: To study the long-term (12 year) survival and quality of life (QOL) in former ICU patients. SETTING: Two hundred and thirty-six ICU admissions from a total of 219 patients treated in a Norwegian University Hospital in 1987. DESIGN AND METHODS: A retrospective analysis of the ICU stays and a prospective observation of survival using available information from the Norwegian Peoples Registry. QOL was studied in survivors in 2000 using the Short Form-36 (SF-36) questionnaire. Survival was compared with available statistics for the general Norwegian population (gender- and age-matched), and QOL was compared with published data from a Norwegian reference population. INTERVENTIONS: None. RESULTS: A total of 106 (48.4 %) patients survived the first 12 years after ICU. Of the non-survivors (113) 66.4 % died within the first year. Two years after discharge the further survival of former ICU patients was 0.763 compared to 0.826 in the general population (difference 0.063 with 95 % CI from -0.007 to 0.134). QOL was significantly less than in the reference population in six of the eight scales of SF-36 (average 82.5%). CONCLUSIONS: Our findings indicate that the long term outcome after ICU is good, with an acceptable QOL and a life expectancy comparable with the general population in survivors 2 years after the ICU stay. PMID- 11497133 TI - Accuracy of a composite score using daily SAPS II and LOD scores for predicting hospital mortality in ICU patients hospitalized for more than 72 h. AB - In most databases used to build general severity scores the median duration of intensive care unit (ICU) stay is less than 3 days. Consequently, these scores are not the most appropriate tools for measuring prognosis in studies dealing with ICU patients hospitalized for more than 72 h. PURPOSE: To develop a new prognostic model based on a general severity score (SAPS II), an organ dysfunction score (LOD) and evolution of both scores during the first 3 days of ICU stay. DESIGN: Prospective multicenter study. SETTING: Twenty-eight intensive care units (ICUs) in France. PATIENTS: A training data-set was created with four ICUs during an 18-month period (893 patients). Seventy percent of the patients were medical (628) aged 66 years. The median SAPS II was 38. The ICU and hospital mortality rates were 22.7% and 30%, respectively. Forty-seven percent (420 patients) were transferred from hospital wards. In this population, the calibration (Hosmer-Lemeshow chi-square: 37.4, P = 0.001) and the discrimination [area under the ROC curves: 0.744 (95 % CI: 0.714-0.773)] of the original SAPS II were relatively poor. A validation data set was created with a random panel of 24 French ICUs during March 1999 (312 patients). MEASUREMENTS AND MAIN RESULTS: The LOD and SAPS II scores were calculated during the first (SAPS1, LOD1), second (SAPS2, LOD2), and third (SAPS3, LOD3) calendar days. The LOD and SAPS scores alterations were assigned the value "1" when scores increased with time and "0" otherwise. A multivariable logistic regression model was used to select variables measured during the first three calendar days, and independently associated with death. Selected variables were: SAPS II at admission [OR: 1.04 (95 % CI: 1.027 1.053) per point], LOD [OR: 1.16 (95 % CI: 1.085-1.253) per point], transfer from ward [OR: 1.74 (95 % CI: 1.25-2.42)], as well as SAPS3-SAPS2 alterations [OR: 1.516 (95 % CI: 1.04-2.22)], and LOD3-LOD2 alterations [OR: 2.00 (95 % CI: 1.29 3.11)]. The final model has good calibration and discrimination properties in the training data set [area under the ROC curve: 0.794 (95 % CI: 0.766-0.820), Hosmer Lemeshow C statistic: 5.56, P = 0.7]. In the validation data set, the model maintained good accuracy [area under the ROC curve: 0.826 (95 % CI: 0.780-0.867), Hosmer-Lemeshow C statistic: 7.14, P = 0.5]. CONCLUSIONS: The new model using SAPS II and LOD and their evolution during the first calendar days has good discrimination and calibration properties. We propose its use for benchmarking and evaluating the over-risk of death associated with ICU-acquired nosocomial infections. PMID- 11497134 TI - The Critical Nursing Situation Index for safety assessment in intensive care. AB - OBJECTIVE: The assessment of critical nursing situations can be a valuable tool in the detection of weak elements in the safety of patients and the quality of care in the ICU. A critical nursing situation can be defined as any observable situation, which deviates from good clinical practice and which may potentially lead to an adverse event. The aim of our study was to establish the feasibility, reliability and validity of the Critical Nursing Situation Index (CNSI) as a tool for assessing the safety and the quality of nursing in the ICU. DESIGN: We described the deviations from standards and protocols in daily ICU nursing care, selected those with an implicit, clear risk for the patients and translated them into explicitly observable items. If an item was applicable during observation of the ICU practice, a critical nursing situation could be recorded as either true or false. The reliability of the CNSI was defined in terms of inter-observer agreement. The validity was assessed by exploring the relationship between the nursing time available (more or less than 30 min per patient per hour) and the incidence of critical nursing situations. SETTING: The study was performed in the ICU of a teaching hospital (30 IC beds) in which all disciplines, including cardiothoracic surgery and neurosurgery, were represented. PATIENTS: The CNSI was randomly applied to 83 ICU patients over a period of 3 months (200 times). MEASUREMENTS AND RESULTS: The reliability of the index was substantial (Kappa values in the range > or =0.70 to > 0.80). In terms of validity, less nursing time resulted in more critical situations (pooled relative risk (RR) 1.36; 95% confidence limits 1.11/1.67). CONCLUSION: The CNSI is simple to use and has encouraging metric properties, whereas the assessments are closely related to direct patient care. Moreover, the CNSI provides a tool for safety assessment by monitoring potentially dangerous situations that are generally regarded as needing to be avoided. PMID- 11497135 TI - Assessing the effectiveness of critical pathways on reducing resource utilization in the surgical intensive care unit. AB - OBJECTIVES: To evaluate the effectiveness of procedure-specific surgical critical pathways on reducing resource utilization in a university surgical intensive care unit (ICU). DESIGN AND SETTING: Prospective cohort study in a university surgical ICU. PATIENTS: 194 patients, accounting for 255 patient days, sampled on randomly selected days over a 12-month period of time. MEASUREMENTS AND RESULTS: The primary outcomes of this study were pathway eligibility and laboratory utilization. Patients were eligible for a procedure-specific pathway in 34% of patient days identified, and the patient's clinical course was "on" pathway in 22% of patient days. Of those "on" the pathway, 54% had a pathway present in the chart and 32% of these included documentation of the patient's clinical course. Thus in 78% of the patient days the patient was either not eligible for a critical pathway or the patient's clinical course was "off" pathway. In those patients "on" the pathway 46 % did not have a pathway present in the chart. Being on a critical pathway did not reduce laboratory utilization. Laboratory utilization did not vary between patients "on" and "off" the pathway (19.1 +/- 11.3 laboratory tests/patient day versus 20.4 +/- 5.7 laboratory tests/patient day). Predicted laboratory utilization by the pathway was 5.6 laboratory tests/patient day. By reducing actual laboratory utilization to that predicted by the critical pathway we would reduce laboratory utilization at our institution by $1.2 million per year. CONCLUSIONS: Procedure-specific surgical critical pathways are not an effective tool for reducing resource utilization in our ICU. Most of our patients were not eligible for an available pathway, and those who were eligible and were "on" the pathway did not appear to have laboratory utilization guided by the pathway. Future initiatives need to explore other means such as ICU specific care processes to reduce resource utilization in the ICU. PMID- 11497136 TI - Intermittent versus continuous renal replacement therapy in the ICU: impact on electrolyte and acid-base balance. AB - OBJECTIVES: The maintenance of normal serum sodium, potassium and bicarbonate concentrations is a therapeutic goal of renal replacement therapy (RRT) in acute renal failure (ARF). The aim of this study was to determine whether this goal is best achieved with intermittent hemodialysis (IHD) or continuous venovenous hemodiafiltration (CVVHDF). DESIGN: Retrospective controlled study. SETTING: Tertiary intensive care unit. PATIENTS: Consecutive patients with ARF treated with IHD (n = 47) or CVVHDF (n = 49). INTERVENTIONS: Measurement of daily morning sodium, potassium and bicarbonate concentrations after the initiation of RRT for up to 2 weeks of treatment. MEASUREMENTS AND RESULTS: Before RRT, abnormal (high or low) values were frequently observed for sodium (42.6% vs 39.6%; NS) potassium (23.4% vs 45.8%; NS) and bicarbonate (63.2 % vs 54.3 %; NS). After treatment, however, CVVHDF, but not IHD, significantly increased mean sodium concentrations (p = 0.0001). CVVHDF was also more likely to normalize the serum sodium than IHD (76.2 % vs 47.8% p = 0.0001). The mean potassium concentrations of both groups significantly decreased (p = 0.019 vs p = 0.0075, difference: NS). However, CVVHDF more frequently reduced the incidence of hypokalemia (1.9 % vs 7.1%, p = 0.0006). CVVHDF but not IHD significantly increased mean bicarbonate concentrations (p = 0.016) in the first 48 h, and more frequently normalized them (71.5% vs 59.2, p = 0.0073). CONCLUSIONS: Serum sodium and potassium, and arterial bicarbonate, concentrations are frequently abnormal in ARF patients before and during renal replacement. Normalization of these values, however, is achieved more frequently with CVVHDF than with IHD. PMID- 11497137 TI - Is a short trial of prone positioning sufficient to predict the improvement in oxygenation in patients with acute respiratory distress syndrome? AB - OBJECTIVE: To determine whether a 1-h trial of prone positioning is sufficient to identify responders. DESIGN: Prospective clinical cohort study in a medico surgical ICU in a teaching hospital. PATIENTS: 49 patients with acute respiratory distress syndrome. INTERVENTIONS: A 6-h period of prone positioning. MEASUREMENTS AND RESULTS: Baseline measurements (blood gas analysis and respiratory parameters) were evaluated in supine position just prior to turning the patients prone. Measurements were then repeated 1 h after the beginning of prone positioning (PP1h) and at the end of the 6-h period of prone positioning (PP6h). The last measurements were performed 1 h after repositioning the patients supine. Prone position induced an increase in the PaO2/FIO2 ratio (p < 0.001). A response (increase in PaO2/FIO2 ratio of at least 20 % at PP1h and/or at PP6h) was observed in 37 of 49 patients (76%). Twenty-seven of these patients (73%) were responders at PP1h while 10 (27%) were responders only at PP6h- In all, two thirds of the patients were considered persistent responders. However, whereas the PaO2/FIO2 ratio decreased significantly 1 h after repositioning the fast responders supine, the PaO2/ FIO2 ratio remained unchanged after repositioning slow responders. CONCLUSIONS: A short-term trial of prone positioning does not appear a sufficient method to identify patients who would benefit from the postural treatment. PMID- 11497138 TI - Efficacy of thrombolysis in patients with acute myocardial infarction requiring cardiopulmonary resuscitation. AB - OBJECTIVE: To evaluate the efficacy and safety of systemic thrombolysis administered to resuscitated patients after cardiac arrest (CA) due to an acute myocardial infarction (AMI), through a study of their mortality and haemorrhagic complications. DESIGN: We studied a retrospective cohort of patients with ischaemic heart disease gathered from the database of the Spanish multi-centre project "Analysis of Delay in AMI" (ARIAM). SETTING: Intensive care (ICU) and coronary care (CCU) units of 77 Spanish hospitals. PATIENTS AND PARTICIPANTS: The study period was from 1 January 1995 to 1 January 2000, when 22,922 patients were included in the ARIAM database register; 13,704 were diagnosed with AMI and we studied 303 of these AMI patients admitted after resuscitation for CA. MEASUREMENTS AND RESULTS: Of the 303 patients studied, 228 were male (75.25%); the mean age was 64.57 +/- 12.48 years. Systemic thrombolysis was administered to 67 patients (group I) and the remaining 236 patients were managed without this treatment (group II). The ICU/CCU mortality rate of the series was 39.93 % (121 patients); that of group I was 17.91% (12 patients) and that of group II 46.18% (109 patients) (P < 0.00001). Group I required less mechanical ventilation (group I, 42.85% vs group II, 80.76 %; P < 0.00001) and fewer cardiopulmonary resuscitation attempts (33.34% vs 60.98%, P < 0.0001). Group I also showed a lower incidence of cardiogenic shock (14.28% vs 39.01%, P < 0.0001) and anoxic encephalopathy (8.62% vs 39.89% P = 0.006). There were no fatal haemorrhagic complications in either group. Logistic regression analysis showed the administration of thrombolysis to be an independent variable that protected against mortality. CONCLUSIONS: The administration of thrombolysis to patients with AMI who require resuscitation may be efficacious in reducing mortality and is safe, with no increase in haemorrhagic complications. PMID- 11497139 TI - The interpretation of train-of-four monitoring in intensive care: what about the muscle site and the current intensity? AB - OBJECTIVES: To investigate the effect of current intensity and choice of the stimulated muscle group on train-of-four (TOF) interpretation in the intensive care unit (ICU). DESIGN AND SETTING: Intervention study in a surgical intensive care unit. PATIENTS: 13 ventilated patients requiring prolonged muscle relaxation. MEASUREMENTS AND RESULTS: Prior to blockade TOF responses of left and right orbicularis oculi, adductor pollicis, and plantar flexors were recorded by setting the current intensity at 20, 40, 60, and 80 mA. The minimal current intensity (MCI) providing a supramaximal response was then identified for each muscle. Cisatracurium was then infused aiming to continuously observe a TOF at 2/4 on the left orbicularis oculi at 40 mA. The responses to TOF on all the muscle sites were further recorded at 40, 60, and 80 mA when the endpoint was reached for the first time, and after a 48-h infusion. After cessation of infusion the delay to observe 4/4 responses at TOF was recorded at each site at 40 mA or at MCI if MCI was above 40 mA. MCI did not differ between muscle groups. When the fixed endpoint was reached for the first time on left orbicularis oculi, the TOF response at 40 mA on right orbicularis oculi differed significantly. In contrast, no difference was observed between left and right sides at 40 mA at the other sites, nor at any sites at 60 and 80 mA. The TOF response on orbicularis oculi (left and right sides together) was different at 40 mA, compared to 60 and 80 mA. TOF responses at orbicularis oculi at 60 or 80 mA significantly differed from responses on adductor pollicis or plantar flexor, orbicularis oculi being less sensitive to cisatracurium than adductor pollicis or plantar flexor. After a 48-h infusion the same differences in sensitivities were observed between the muscle groups. At any current intensity the recovery was slower at adductor pollicis than at orbicularis oculi or plantar flexor. CONCLUSIONS: For a good TOF interpretation in the ICU the current intensity should be tested before onset of blockade. The orbicularis oculi is less sensitive to cisatracurium than adductor pollicis and plantar flexor both at onset and after a prolonged infusion. The recovery from relaxation is faster on orbicularis oculi and plantar flexor than on adductor pollicis. PMID- 11497140 TI - Evaluation of a simple method for minimizing iatrogenic blood loss from discard volumes in critically ill newborns and children. AB - OBJECTIVE: To validate a simple method avoiding discard volumes in pediatric patients with indwelling arterial and venous lines. DESIGN: Zero-discarding was achieved by passive extracorporeal arteriovenous backflow via standard single pressure transducer equipment. We tested backflow distances (10, 20 and 30 cm beyond the sampling port), corresponding to withdrawal volumes of 0.6 ml, 0.8 ml and 1.0 ml, respectively, in comparison to conventional sampling with discard of 0.6 ml. With the backflow technique, the "withdrawal volume" was flushed back to the patient after sampling. We enrolled 120 patients who were allocated to either of the following paired sampling procedures: 10 cm versus conventional, 20 cm versus conventional, 30 cm versus conventional and two paired conventional samples. The order of the sampling was randomly allocated. Bias and precision were determined using Bland-Altman diagrams and algorithms. RESULTS: No appreciable difference was found for blood gases, hemoglobin, potassium and calcium between the backflow technique and conventional sampling. Sodium results and blood glucose showed a bias towards higher values with the backflow technique (mean difference, sodium, 0.9 mmol/l; glucose, mean difference 0.5 mmol/l, standard deviation 0.44 mmol/l). CONCLUSIONS: The backflow technique provides reliable results for blood gases and electrolytes. However, in patients at risk of hypoglycemia, the backflow method should not be used to monitor blood glucose levels. PMID- 11497141 TI - Myocardial injury in meningococcus-induced purpura fulminans in children. AB - OBJECTIVES: To assess the incidence of myocardial ischemia in meningococcus induced purpura fulminans in pediatric patients, to compare troponin I (cTnI) levels with changes in electrocardiogram (ECG) and to evaluate whether cTnI is related to myocardial function and contractility, to severe acquired anticoagulant deficiency and to the severity of disease. METHODS: Twenty-two patients with acute meningococcemia, supported with inotropes or vasoactive agents, were studied, Blood samples for the determination of serum cTnI and conventional myocardial ischemia and coagulopathy markers were drawn daily. Measurements of cardiac index (CI), ejection (EF) and shortening fractions (SF) and ECGs were performed daily. RESULTS: The Leclerc score, the Neisseria sepsis index (NESI) and the pediatric risk of mortality (PRISM) score predicted a mean mortality rate of 34%, 27% and 23%, respectively. Four patients died (18%). Five patients (23 %) presented with myocardial ischemia. Their ECG ischemic changes were associated with pathologically high cTnI levels (1.93 +/- 0.13 vs 0.18 +/- 0.08 ng/ml, p < 0.001 for patients with or without ischemic changes) and depressed myocardial contractility (mean difference +/- SE -14 +/- 5%, p = 0.01, for the EF and -7.4 +/- 3, p < 0.02, for the SF). High cTnI values were significantly correlated to low protein C (PC) (p < 0.0001), factor VIII (p < 0.04) and antithrombin III (AIII, p = 0.01) levels, but not to the PRISM, Leclerc or the NESI scores. Means of AIII, VII, and especially of VIII, and PC, were significantly lower in ischemic than in non-ischemic patients, although severity scoring systems and inotropic support did not differ between the two groups. Survivors tended to significantly higher PC (p < 0.01) and factor VIII levels (p = 0.001) than non-survivors and, also, to lower levels of cTnI (p = 0.05) and CPK MB (p < 0.05), while in meningococcal shock. CONCLUSIONS: The incidence of myocardial ischemia is increased in acute meningococcemia in pediatric patients and correlates with myocardial dysfunction. High cTnI is associated with severe coagulopathy, but not with clinical prognostic scores or inotropic support. Early recognition of myocardial injury, myocardial support and early replacement therapy with PC, AIII, factor VIII or fibrinogen might improve outcome in acute meningococcemia in children. PMID- 11497142 TI - Somatostatin: a new therapeutic option for the treatment of chylothorax. AB - BACKGROUND: The standard treatment of chylothorax in pediatric intensive care today includes conservative therapy with fat-free nutrition, total parenteral nutrition and, if this is not successful, operative treatment (pleurodesis, ligation of the duct, pleuroperitoneal shunt). PATIENTS: We describe four patients who were not in a suitable condition for operative treatment and who were treated with continuous infusion of somatostatin. RESULTS: In three patients, chylothorax ceased with the continuous somatostatin infusion without side effects. One patient was treated without success. CONCLUSIONS: Somatostatin is a therapeutic option for treatment of chylothorax and could reduce surgical intervention and hospitalization time, as well as allow earlier enteral feeding. PMID- 11497143 TI - Ketamine and midazolam kinetics during continuous hemodiafiltration in patients with multiple organ dysfunction syndrome. AB - OBJECTIVE: To assess the effect of continuous hemodiafiltration (CHDF) on ketamine and midazolam kinetics in multiple organ dysfunction syndrome (MODS). DESIGN AND SETTING: Consecutive clinical study in a general intensive care unit of a university hospital. PATIENTS: Twelve adult patients with MODS requiring CHDF. MEASUREMENTS AND RESULTS: A total of 68 samples were collected during CHDF for ketamine, norketamine, and midazolam assays. The clearance values for ketamine and norketamine were 10.8 +/- 6.6 and 10.9 +/- 11.5 ml/min and their daily extractions were 21.4 +/- 7.1 and 10.2 +/- 11.5 mg/day, respectively. Midazolam was not eliminated through the filter during CHDF. There were no changes in Ramsay Sedation Score or Glasgow Coma Scale during CHDF. CONCLUSIONS: Small fractions of ketamine and norketamine were eliminated during CHDF in MODS. Midazolam was not eliminated during CHDF. CHDF did not affect the sedation using ketamine and midazolam even in MODS patients. PMID- 11497144 TI - The role of cell suicide or apoptosis in the pathophysiology of acute lung injury. PMID- 11497145 TI - Tick bite induced respiratory failure. Diaphragm palsy in Lyme disease. PMID- 11497146 TI - Hypokalemia causing rhabdomyolysis and precordialgia. PMID- 11497147 TI - Are cytokines and adhesion molecules useful parameters to differentiate bacterial from fungal ventriculitis? PMID- 11497149 TI - Toothache leading to emergency cardiac surgery. PMID- 11497148 TI - Silent transmural myocardial ischemia during septic shock in a patient with normal coronary arteries. PMID- 11497150 TI - Congenital diaphragmatic hernia: outcome in small centres. PMID- 11497151 TI - Cardiopulmonary effects of nitric oxide inhalation and methylene blue injection in hepatopulmonary syndrome. PMID- 11497152 TI - Increased troponin in patients with sepsis and septic shock: myocardial necrosis or reversible myocardial depression? PMID- 11497153 TI - Outcome prediction in intensive care. Solving the paradox. PMID- 11497154 TI - Elevation of troponin I in sepsis and septic shock. AB - OBJECTIVE: To detect myocardial damage in severe systemic inflammation by cTnI measurements in patients without acute coronary syndromes. DESIGN: Prospective case control study. SETTING: Tertiary referral center. PARTICIPANTS: Twenty patients with sepsis, septic shock, and systemic inflammatory response syndrome (SIRS) were examined and compared to controls without coronary artery disease or myocarditis. MEASUREMENTS AND RESULTS: cTnI levels were assessed in patients with SIRS, sepsis, and septic shock. Eight patients (two female/six male) suffered from septic shock, nine (three female/six male) from sepsis without shock, and three (three male) from SIRS. Seventeen patients (85%) showed elevated cTnI (median 0.57 microg/l; 0.17-15.4), whereas no patient in the control group showed elevated cTnI (P < 0.0001). Six patients (30%),--three with septic shock and three with sepsis--died during hospitalization, five of them with elevated cTnI. Four out of five autopsies showed normal coronary arteries. Coronary angiography, autopsy, and stress echocardiography ruled out significant coronary artery disease in ten cTnI-positive patients (59%). In 41 % of cTnI-positive patients, Streptococcus pneumoniae could be cultured, whereas no cTnI-negative or control patient showed signs of infection due to S. pneumoniae. CONCLUSION: Cardiac troponin I was elevated in 85% of patients with sepsis, septic shock or SIRS in our study. A high percentage showed infection caused by S. pneumoniae. In what way microorganisms cause cTnI elevations is not yet understood. PMID- 11497155 TI - Early prognosis in severe sepsis via analyzing the monocyte immunophenotype. AB - OBJECTIVE: To analyze the early discriminative predictive information regarding the immunophenotype components of patients with sepsis, and its potential use as a prognosis tool. DESIGN: Observational prospective clinical study. SETTING: Intensive care unit (ICU) in a University Hospital. PATIENTS: Thirty-five patients admitted with severe sepsis. MEASUREMENTS: Analysis of peripheral blood on admission and 48 h later of the absolute white cell count and the immunophenotype of lymphocyte (CD3, CD3-HLADR, CD4, CD8, CD4/CD8 ratio, CD19, and CD25) and monocyte (CD13, CD13-HLADR, CD14, CD14-HLADR, CD13-CD14, and CD4) subpopulations. RESULTS: Due to its high correlation, the immunophenotypic profile studied at admission and 48 h later showed the same prognosis power regardless of the time of performance. The univariate analysis between groups (survival versus death) confirmed the prognostic significance of the total monocyte count and its subpopulations; significant differences were observed from the beginning only in the CD19 lymphocyte subpopulation. Multivariate analysis was performed using logistic regression with survival as the dependent variable. The final model comprised monocytes beta = 0.002 (P = 0.025) and CD13-HLADR beta = 0.016 (P = 0.029). The monocytes receiver operating characteristic (ROC) area obtained was 0.819 (confidence interval 0.663-0.976 at 95 %), the CD13-HLADR ROC area was 0.810 (confidence interval 0.658-0.963), and the monocytes + CD13-HLADR ROC area was 0.918 (confidence interval 0.807-1.000). CONCLUSIONS: A single blood sample test obtaining the absolute monocyte and CD13-HLADR subpopulation count in the first days of admission could contribute to simplifying the classification of patients with severe sepsis into high- and low-mortality risk. PMID- 11497156 TI - High-volume haemofiltration in human septic shock. AB - OBJECTIVE: To evaluate whether high volume haemofiltration improves haemodynamics and affects serum cytokine and complement concentrations in human septic shock. DESIGN AND SETTING: Randomized cross-over clinical trial in a tertiary intensive care unit. PATIENTS: Eleven patients with septic shock and multi-organ failure. INTERVENTIONS: Patients were assigned to either 8 h of high-volume haemofiltration (HVHF; 6 l/h) or 8 h of standard continuous veno-venous haemofiltration (CVVH; 1 l/h) in random order. MEASUREMENTS AND MAIN RESULTS: We measured changes in haemodynamic variables, dose of norepinephrine required to maintain a mean arterial pressure greater than 70 mmHg and plasma concentrations of complement anaphylatoxins and several cytokines. An 8-h period of HVHF was associated with a greater reduction in norepinephrine requirements than a similar period of CVVH (median reduction: 10.5 vs. 1.0 microg/min; p = 0.01; median percentage reduction: 68 vs. 7%; p = 0.02). Both therapies were associated with a temporary reduction (p < 0.01) in the plasma concentration of C3a, C5a, and interleukin 10 within 2 h of initiation. HVHF was associated with a greater reduction in the area under the curve for C3a and C5a (p < 0.01). The concentration of the measured soluble mediators in the ultrafiltrate was negligible. CONCLUSIONS: HVHF decreases vasopressor requirements in human septic shock and affects anaphylatoxin levels differently than standard CVVH. PMID- 11497157 TI - Massive elevation of procalcitonin plasma levels in the absence of infection in kidney transplant patients treated with pan-T-cell antibodies. AB - OBJECTIVE: To determine the value of procalcitonin (PCT) monitoring in transplant patients receiving pan-T-cell antibody therapy. DESIGN: Retrospective clinical study. SETTING: A collaborative study between the Institute of Medical Immunology, the Department of Nephrology and Internal Intensive Care, both Charite, Humboldt University Berlin, and the Department of Laboratory Medicine, Friedrichshain Hospital, Berlin, Germany. PATIENTS AND INTERVENTIONS: Thirty-one patients were included in the study: 8 kidney transplant patients with acute rejection episodes, 5 receiving OKT3 monoclonal antibody therapy, 3 receiving steroid bolus therapy; 21 patients undergoing renal transplantation, 11 receiving ATG perioperatively, 10 without ATG administration; 2 patients undergoing renal transplantation and receiving anti-IL-2R mAb. MEASUREMENTS AND RESULTS: Procalcitonin (PCT) and tumor necrosis factor (TNF) alpha plasma levels were measured in infection-free transplant patients treated with the pan-T-cell antibodies ATG or OKT3. We found PCT plasma concentrations up to 600 ng/ml (reference < 0.5 ng/ ml), which are comparable to those seen in severe sepsis. Increases in TNF-alpha plasma levels preceded the rises in PCT. After peaking on day 1 of therapy the PCT plasma concentrations returned to normal values independently of further antibody administration. In contrast, steroid bolus therapy or anti-interleukin 2 receptor mAb administration did not increase plasma PCT or TNF-alpha levels. CONCLUSIONS: PCT monitoring for evaluating infectious complications in kidney transplant patients must be very careful during pan-T cell antibody therapy. PMID- 11497158 TI - Evaluation of the logistic organ dysfunction system for the assessment of organ dysfunction and mortality in critically ill patients. AB - OBJECTIVES: To evaluate the performance of the logistic organ dysfunction (LOD) system for the assessment of morbidity and mortality in multiple organ dysfunction/failure (MOD/F) in an independent database and to evaluate the use of sequential LOD measurements for the prediction of outcome. DESIGN AND SETTING: Prospective, multicentric cohort study in 13 adult medical, surgical, and mixed intensive care units (ICUs) in Austria. PATIENTS: A total of 2,893 consecutive admissions to the ICUs. MEASUREMENTS AND MAIN RESULTS: Patient vital status at ICU and hospital discharge was recorded. Univariate analysis showed that the LOD was able to distinguish between survivors and nonsurvivors (2 vs. 6 median score). Within organ systems, higher levels of the severity of organ dysfunction were consistently associated with higher mortality. For the prediction of hospital mortality, the original prognostic LOD model did not perform well in our patients, as indicated by the goodness-of-fit C statistic. Using multiple logistic regression we developed a prognostic model with a satisfactory fit in our patients. The integration of further measurements during the ICU stay increased discrimination but not calibration. CONCLUSIONS: The LOD system is well correlated well with the numbers and levels of organ dysfunctions and discriminates well between survivors and nonsurvivors. It can thus be used to quantify the baseline severity of organ dysfunction. Moreover, after customization of the predictive equation the LOD predicted hospital mortality in our patients with high precision. It thus provides a combined measure of morbidity and mortality for critically ill patients with MOD/F. PMID- 11497159 TI - Mortality after discharge from intensive care: the impact of organ system failure and nursing workload use at discharge. AB - OBJECTIVES: Mortality after ICU discharge accounts for approx. 20-30% of deaths. We examined whether post-ICU discharge mortality is associated with the presence and severity of organ dysfunction/failure just before ICU discharge. PATIENTS AND METHODS: The study used the database of the EURICUS-II study, with a total of 4,621 patients, including 2,958 discharged alive to the general wards (post-ICU mortality 8.6%). Over a 4-month period we collected clinical and demographic characteristics, including the Simplified Acute Physiology Score (SAPS II), Nine Equivalents of Nursing Manpower Use Score, and Sequential Organ Failure Assessment (SOFA) score. RESULTS: Those who died in the hospital after ICU discharge had a higher SAPS II score, were more frequently nonoperative, admitted from the ward, and had stayed longer in the ICU. Their degree of organ dysfunction/failure was higher (admission, maximum, and delta SOFA scores). They required more nursing workload resources while in the ICU. Both the amount of organ dysfunction/failure (especially cardiovascular, neurological, renal, and respiratory) and the amount of nursing workload that they required on the day before discharge were higher. The presence of residual CNS and renal dysfunction/failure were especially prognostic factors at ICU discharge. Multivariate analysis showed only predischarge organ dysfunction/failure to be important; thus the increased use of nursing workload resources before discharge probably reflects only the underlying organ dysfunction/failure. CONCLUSIONS: It is better to delay the discharge of a patient with organ dysfunction/failure from the ICU, unless adequate monitoring and therapeutic resources are available in the ward. PMID- 11497160 TI - Comparing approximations to spatio-temporal models for epidemics with local spread. AB - Analytical methods for predicting and exploring the dynamics of stochastic, spatially interacting populations have proven to have useful application in epidemiology and ecology. An important development has been the increasing interest in spatially explicit models, which require more advanced analytical techniques than the usual mean-field or mass-action approaches. The general principle is the derivation of differential equations describing the evolution of the expected population size and other statistics. As a result of spatial interactions no closed set of equations is obtained. Nevertheless, approximate solutions are possible using closure relations for truncation. Here we review and report recent progress on closure approximations applicable to lattice models with nearest-neighbour interactions, including cluster approximations and elaborations on the pair (or pairwise) approximation. This study is made in the context of an SIS model for plant-disease epidemics introduced in Filipe and Gibson (1998, Studying and approximating spatio-temporal models for epidemic spread and control, Phil. Trans. R. Soc. Lond. B 353, 2153-2162) of which the contact process [Harris, T. E. (1974), Contact interactions on a lattice, Ann. Prob. 2, 969] is a special case. The various methods of approximation are derived and explained and their predictions are compared and tested against simulation. The merits and limitations of the various approximations are discussed. A hybrid pairwise approximation is shown to provide the best predictions of transient and long-term, stationary behaviour over the whole parameter range of the model. PMID- 11497161 TI - Diffusion induced oscillatory insulin secretion. AB - Oscillatory secretion of insulin has been observed in many different experimental preparations. Here we examine a mathematical model for in vitro insulin secretion from pancreatic beta cells in a flow-through reactor. The analysis shows that oscillations result because of an important interplay between flow rate of the reactor and insulin diffusion. In particular, if the ratio of flow rate to volume of the reaction bed is too large, oscillations are eliminated, in contradiction to the conclusions of Maki and Keizer (L. W. Maki and Keizer J. Mathematical analysis of a proposed mechanism for oscillatory insulin secretion in perifused HIT-15 cells. Bull. Math. Biol., 57(1995), 569-591). Furthermore, with reasonable numbers for the experimental parameters and the diffusion of insulin, the model equations do not exhibit oscillations. PMID- 11497162 TI - Spontaneous signal generation in living cells. AB - Living cells often generate signals spontaneously in the absence of external stimuli. Those signals play an important role in their tactic behaviors. This paper presents a theoretical treatment on the mechanism of spontaneous signal generation. The mechanism consists of two steps: (1) production of the basic fluctuation of the intracellular electric potential due to the open-closed fluctuation of the gates of ion channels and (2) generation of a spike-like fluctuation of potential depending on the positive shift of the basic fluctuation. The first step is described by an equation of the Langevin type, where the random force is proportional to the circulating ion current across the membrane; the average of the square of the random force is proportional to the rate of free-energy consumption by the current. The second step is described by a rate equation of transition of field-sensitive channel gates which contains the fluctuating electric field in the exponential term. There, the fluctuation has a nonlinear effect. Such a two-step process may work in various kinds of living cells. The presence of circulating ion current in the resting state is a most important key. Some cells may be quiet and some cells may be active to generate spontaneous signals. PMID- 11497163 TI - How predation can slow, stop or reverse a prey invasion. AB - Observations on Mount St Helens indicate that the spread of recolonizing lupin plants has been slowed due to the presence of insect herbivores and it is possible that the spread of lupins could be reversed in the future by intense insect herbivory [Fagan, W. F. and J. Bishop (2000). Trophic interactions during primary sucession: herbivores slow a plant reinvasion at Mount St. Helens. Amer. Nat. 155, 238-251]. In this paper we investigate mechanisms by which herbivory can contain the spatial spread of recolonizing plants. Our approach is to analyse a series of predator-prey reaction-diffusion models and spatially coupled ordinary differential equation models to derive conditions under which predation pressure can slow, stall or reverse a spatial invasion of prey. We focus on models where prey disperse more slowly than predators. We comment on the types of functional response which give such solutions, and the circumstances under which the models are appropriate. PMID- 11497164 TI - Stability of a diverse immunological memory is determined by T cell population dynamics. AB - The correlation between properties of the T cell memory pool and the two regulatory mechanisms of cell death (apoptosis) and memory entry (differentiation) is investigated mathematically. Apoptosis of T cells occurs at the end of an immune response, removing unwanted activated T cells. T cells escaping apoptosis enter the memory pool composed of T cells specific for previously encountered antigens. We find that the relative efficiencies of these two pathways determine the clonal distribution and the long-term stability of the memory pool by regulating the number of new entries. The main result presented in this paper is that immunological memory of previously encountered pathogens cannot be erased by either severe or repeat infections with a particular pathogen (the diversity of the memory pool is ensured) only if apoptosis and/or memory differentiation are regulated by population dependent processes. Furthermore, vaccination properties are improved significantly by population dependent mechanisms and our mathematical analysis reveals that the T cell population must communicate with other parts of the immune system to ensure optimal performance of immunological memory. PMID- 11497165 TI - Adaptive evolution on neutral networks. AB - We study the evolution of large but finite asexual populations evolving in fitness landscapes in which all mutations are either neutral or strongly deleterious. We demonstrate that despite the absence of higher fitness genotypes, adaptation takes place as regions with more advantageous distributions of neutral genotypes are discovered. Since these discoveries are typically rare events, the population dynamics can be subdivided into separate epochs, with rapid transitions between them. Within one epoch, the average fitness in the population is approximately constant. The transitions between epochs, however, are generally accompanied by a significant increase in the average fitness. We verify our theoretical considerations with two analytically tractable bitstring models. PMID- 11497166 TI - Modeling and analysis of a virus that replicates selectively in tumor cells. AB - Replication-competent viruses have shown considerable promise in overcoming the inefficient gene transduction experienced by traditional gene therapy approaches to cancer treatment. The viruses infect tumor cells and replicate inside them, eventually causing lysis. Virus particles released during lysis are then able to infect other tumor cells, and, in this way, continuous rounds of infection and lysis allow the virus to spread throughout the tumor. Motivated by this novel cancer treatment, we formulate and analyse a system of partial differential equations that is essentially a radially-symmetric epidemic model embedded in a Stefan problem. We compare three, alternative virus-injection strategies: a fixed fraction of cells pre-infected with the virus are introduced throughout the entire tumor volume, within the tumor core, or within the tumor rim. For all three injection methods, simple and accurate conditions that predict whether the virus will control the tumor are derived. PMID- 11497167 TI - Iteroparous reproduction strategies and population dynamics. AB - Asymptotic relationships between a class of continuous partial differential equation population models and a class of discrete matrix equations are derived for iteroparous populations. First, the governing equations are presented for the dynamics of an individual with juvenile and adult life stages. The organisms reproduce after maturation, as determined by the juvenile period, and at specific equidistant ages, which are determined by the iteroparous reproductive period. A discrete population matrix model is constructed that utilizes the reproductive information and a density-dependent mortality function. Mortality in the period between two reproductive events is assumed to be a continuous process where the death rate for the adults is a function of the number of adults and environmental conditions. The asymptotic dynamic behaviour of the discrete population model is related to the steady-state solution of the continuous-time formulation. Conclusions include that there can be a lack of convergence to the steady-state age distribution in discrete event reproduction models. The iteroparous vital ratio (the ratio between the maximal age and the reproductive period) is fundamental to determining this convergence. When the vital ratio is rational, an equivalent discrete-time model for the population can be derived whose asymptotic dynamics are periodic and when there are a finite number of founder cohorts, the number of cohorts remains finite. When the ratio is an irrational number, effectively there is convergence to the steady-state age distribution. With a finite number of founder cohorts, the number of cohorts becomes countably infinite. The matrix model is useful to clarify numerical results for population models with continuous densities as well as delta measure age distribution. The applicability in ecotoxicology of the population matrix model formulation for iteroparous populations is discussed. PMID- 11497168 TI - The enigma of short-segment Barrett's esophagus. PMID- 11497169 TI - Seroepidemiology of hepatitis A infection in India: changing pattern. AB - OBJECTIVE: Recent changes in the epidemiology of hepatitis A virus (HAV) infection and the availability of effective vaccines have renewed interest in this infection. We determined the age-related prevalence of anti-HAV antibodies in India and looked for differences by known risk factors for HAV infection. METHODS: In this prospective study, serum samples obtained from 1612 subjects aged 1 to 60 at six centers in five cities (Calcutta, Cochin, Indore, Jaipur and Patna) during the period February to August 1998 were tested for anti-HAV antibodies. Demographic and socio-economic information was obtained by questionnaire. RESULTS: The overall seroprevalence rate was 65.9%, varying from 26.2% to 85.3% in various cities; there was no difference between males and females. Seropositivity increased with age from 52.2% in the 1-5 year age group to 80.8% in those aged 16 years or more. Seroprevalence rates were significantly lower in those aged 1-5 years compared with other age groups (p<0.0001). There was no difference in seroprevalence between those with monthly family income Rs 5001. Multivariate analysis showed that anti-HAV seroprevalence varied significantly by source of water supply, being highest when the supply was municipal. CONCLUSION: Our results indicate an epidemiological pattern of intermediate endemicity. This finding has public health implications as it indicates that a significant proportion of the Indian adolescent and adult population is at risk of HAV infection. PMID- 11497170 TI - Surgical anatomy of retrohepatic inferior vena cava and hepatic veins: a quantitative assessment. AB - BACKGROUND: Accurate knowledge of the surgical anatomy of the retrohepatic inferior vena cava (IVC) and hepatic veins is necessary for hepatic surgery. METHODS: Lengths of different segments of retrohepatic IVC and their diameters, and prevalence of various types of ramification and lengths of different hepatic veins, were noted in 100 disease-free human livers during autopsy. RESULTS: The mean lengths of the IVC from entry into atrium to diaphragmatic hiatus, from the hiatus to the upper margin of right hepatic vein, between the upper margins of the right hepatic vein and the right suprarenal vein, from right suprarenal vein to the lowermost dorsal hepatic vein, and from the lower-most dorsal hepatic vein to the right renal vein were 29.1 mm, 8.6 mm, 40.6 mm, 28.6 mm and 33.7 mm, respectively. The mean diameter of IVC at the diaphragmatic level was 30.1 mm. The commonest ramification pattern of the hepatic veins was type I (82%) for the right hepatic vein, type II (63%) for the middle and left hepatic veins, and type II (55%) for the caudate veins. In 96% of cases the middle and left hepatic veins formed a common trunk. In a majority of cases, the diameters of the right and left hepatic veins were between 7 mm and 12 mm. No gender differences were found. CONCLUSION: This study provides an anatomical perspective for various hepatic surgical techniques. PMID- 11497171 TI - 14C-urea breath test for assessment of gastric Helicobacter pylori colonization and eradication. AB - BACKGROUND AND OBJECTIVE: Urea breath test (UBT) is a reliable noninvasive technique for detecting gastric Helicobacter pylori colonization. 14C isotope based test requires simple equipment and is inexpensive. We studied the utility of 14C-UBT in diagnosis of gastric H. pylori infection. METHODS: Presence of H. pylori was studied using antral histology and culture in patients with rapid urease test (RUT)-positive peptic ulcer. 14C-UBT was performed using a 185-kBq dose. Radioactivity in 15-min breath samples was measured using a beta scintillation counter and result expressed as % dose recovered/mmol CO2. H. pylori was considered positive when any two tests were positive. All tests were repeated one month after completion of H. pylori eradication therapy. RESULTS: Among 41 patients (duodenal ulcer 36, gastric ulcer 5), H. pylori was detected by histology in 23 (56%) and by culture in 27 (66%). Overall, H. pylori was detected in 28 (68%) patients. Follow-up assessment was possible in 28 patients: 26 cleared the infection (all three tests negative). Mean 14C recovery values at 15 minutes associated with H. pylori-positive status were significantly higher (12.3 [SD 6.8] x 10(-3); n=30; p<0.001) than those associated with H. pylori-negative status (2.1 [0.9] x 10(-3); n=26). Using receiver-operating-characteristic analysis of 15-minute 14C recovery values, a cut-off of 6.5x10(-3) gave the best separation of H. pylori-positive and -negative cases. 14C-UBT had 93% sensitivity, 96% specificity and 95% accuracy. CONCLUSION: 14C-UBT appears to be a reliable noninvasive test for diagnosis of H. pylori infection. PMID- 11497172 TI - Prevalence of short segments of specialized columnar epithelium in distal esophagus: association with gastroesophageal reflux. AB - BACKGROUND AND AIMS: Recent studies have reported high prevalence rates of short segments of specialized columnar epithelium (SCE) in the distal esophagus. The association of SCE with gastroesophageal reflux disease is not well established. We studied the prevalence and associations of short segments of SCE in the distal esophagus amongst Indians. METHODS: 271 patients (mean age 36 [14] y; 160 men) undergoing diagnostic upper gastrointestinal endoscopy were interviewed regarding symptoms of gastroesophageal reflux, and history of medications, smoking or chewing tobacco and alcohol ingestion. At endoscopy, presence and grade of esophagitis and hiatus hernia were recorded. One biopsy each was taken from the squamocolumnar junction and 2 cm proximal to it. Biopsies were stained with hematoxylin/eosin and alcian blue/periodic acid-Schiff. The pathologist was blinded to the clinical and endoscopic data. RESULTS: Short segments of SCE in the distal esophagus were present in 16/271 (6%; CI 5.03-6.97) patients. Increasing age (p<0.01), and endoscopic (p<0.01) and histologic (p<0.001) esophagitis were associated with its presence, whereas symptoms of gastroesophageal reflux, smoking, tobacco chewing, use of alcohol or non steroidal anti-inflammatory drugs, and hiatus hernia were not. One patient with SCE had dysplasia. CONCLUSION: Prevalence of short segments of SCE in the distal esophagus amongst Indians is low and is usually associated with inflammation in the esophagus. PMID- 11497173 TI - Protective effect of L-carnitine on gastric mucosal barrier in rats exposed to cold-restraint stress. AB - AIM: The protective effect of L-carnitine on stress-induced gastric mucosal injury was investigated in rats exposed to cold-restraint stress (CRS). METHODS: The animals were divided into four groups. Groups 1 and 3 received saline by intragastric gavage for 10 days. Groups 2 and 4 received L-carnitine (50 mg/Kg/day) in the same manner. Groups 3 and 4 were exposed to CRS in the form of immobilization at 4 degrees C for 4 h on day 10. Ulcer index, gastric acid secretion and hemoglobin leakage, and gastric mucosal mucin and PGE2 content were measured. RESULTS: In rats exposed to CRS, as compared to control rats (group 1), ulcer index was higher, gastric acid production was lower, hemoglobin leakage into the gastric lumen was increased, and gastric mucosal mucin and PGE content were reduced. L-carnitine treatment prior to CRS led to attenuation of changes in ulcer index, gastric acid secretion, amount of hemoglobin leakage into the gastric lumen and gastric PGE2 content. In rats receiving L-carnitine but not exposed to CRS, gastric acid secretion, mucin and PGE2 content of gastric mucosa were similar to those in control rats. CONCLUSION: L-carnitine decreases CRS induced gastric mucosal injury. PMID- 11497174 TI - Management of the Budd-Chiari syndrome by balloon cavoplasty. AB - BACKGROUND: Obstruction of the suprahepatic inferior vena cava (IVC) by a membrane or stricture is the commonest cause of Budd-Chiari syndrome in the eastern hemisphere. We present our experience with the outcome of balloon cavoplasty in such cases. METHODS: We followed up 40 consecutive cases of Budd Chiari syndrome over seven years. Doppler study of hepatic venous outflow tract (in all cases), liver biopsy (30 cases) and necropsy (two cases) were performed. Balloon cavoplasty was done in selected cases. RESULTS: Of 40 patients with BCS (mean age 35.2 [SD 8.7] years; 26 men) 5, 5 and 30 had fulminant, acute and chronic presentation, respectively. Inferior vena cavography was performed in 32 cases, and showed membranous obstruction of the IVC in 12, segmental occlusion of the IVC in 11 cases, and block in both the IVC and the main hepatic veins in the rest. Successful balloon cavoplasty was done in 18 cases with obstruction of the IVC (membrane or stricture); 15 of them are well over a mean follow up of 56 (14.6) months. Three patients developed restenosis; two of them, treated with redilatation, are doing well, and one died of septicemia and hepatic failure following a surgical bypass. Pressure gradient between the IVC and right atrium decreased significantly after cavoplasty (15.4 [2.8] vs 6.6 [2.0] mmHg; p< 0.001). CONCLUSION: Balloon cavoplasty gave encouraging results in the management of Budd-Chiari syndrome due to membranous obstruction or stricture of the IVC. PMID- 11497175 TI - Non-syndromic paucity of interlobular bile ducts. AB - A 3 1/2-month-old male infant presented with cholestatic jaundice. Peroperative cholangiogram showed a gall bladder and small but patent extrahepatic bile ducts. Liver histology showed paucity of interlobular bile ducts. The child is being treated medically for his symptoms. PMID- 11497176 TI - Synchronous primary malignant neoplasms of colon and bladder. AB - Extracolonic and synchronous malignancies are rare in colorectal carcinomas. We report a 68-year-old man with complaints of rectal bleeding and hematuria. Endoscopic biopsies revealed synchronous adenocarcinoma of the colon and transitional cell carcinoma of the bladder. The patient was started on chemotherapy, and is alive (with disease) nine months later. PMID- 11497177 TI - Massive amebic liver abscess--an unusual presentation. AB - A 40-year-old woman presented with a huge lump in the right half of the abdomen and irregular menses. Ultrasonography showed a cystic lump with septations, extending from the upper abdomen to the pelvis; the right ovary was not seen. On exploration, there was a large cyst arising from the right lobe of the liver; the aspirate was bilious. Since the cyst wall was adherent to retroperitoneal structures, complete excision was not possible. A roux-en-Y loop of jejunum was anastomosed to the cyst wall. Biopsy of the wall showed inflammatory granulation tissue with trophozoites of Entamoeba histolytica. She was treated with metronidazole, and recovered uneventfully. PMID- 11497178 TI - Gastric teratoma: a rare cause of gastrointestinal bleeding. AB - Gastric teratoma is a rare tumor of infancy, almost exclusively found in males. We report a 6-month-old male child with gastric teratoma who presented with gastrointestinal bleeding; he was treated successfully with local excision of the tumor. PMID- 11497179 TI - Impacted denture causing tracheo-esophageal fistula. AB - We report a 35-year-old man with an impacted denture resulting in tracheo esophageal fistula. In view of significant local fibrosis and esophageal stenosis distal to the fistula, he was managed by subtotal esophagectomy and cervical esophagogastric anastomosis. PMID- 11497180 TI - Strongyloides stercoralis eggs in the stools during anticancer therapy. AB - We report a 45-year-old man with granulocytic sarcoma who, following anticancer chemotherapy, developed florid strongyloidiasis, wherein eggs of Strongyloides stercoralis embedded in the exfoliated mucosa were seen in the stools. The eggs were interspersed with numerous rhabditiform larvae. He was treated with albendazole. PMID- 11497181 TI - Recurrent abscess at site of laparoscopic cholecystectomy port due to spilled gallstones. AB - Spillage of gallstones is common during laparoscopic cholecystectomy and may lead to intra-abdominal abscesses and sinus formation. We describe two patients with recurrent abscess at the site of epigastric port due to presence of large spilled stone in the parietes following laparoscopic cholecystectomy. Removal of the stone led to resolution of symptoms. PMID- 11497182 TI - Analysis of post-cholecystectomy bile duct injuries. PMID- 11497183 TI - Need for antibiotics before sclerotherapy in cirrhotics with acute variceal bleeding. PMID- 11497184 TI - Colonic intussusception due to carcinoma in an adolescent. PMID- 11497185 TI - Fascioliasis: a case fom central Uttar Pradesh. PMID- 11497186 TI - Closure of jejuno-colic fistula in Crohn's disease with infliximab. PMID- 11497187 TI - Laragh's lessons in pathophysiology and clinical pearls for treating hypertension. Lesson XXIV: on the major roles of the renin system in the pathogenesis of hypertension and of its sequelae, heart attack, heart failure, kidney failure, and stroke: replies to commonly asked questions. PMID- 11497188 TI - Relationship between 24-H blood pressure and sleep disordered breathing in a normotensive community sample. AB - Sleep disordered breathing (SDB) and hypertension are commonly associated. In this study, we assessed how longitudinal measures of SDB predict a 24-h ambulatory blood pressure monitoring (ABPM) profile. Participants (n = 82) were recruited from a community-based urban (26% African American) sample and included family members of patients with laboratory diagnosed SDB (cases) and family members of neighborhood control subjects evaluated at baseline and at 5 years. Nearly all participants were normotensive and were not receiving therapy for SDB. During both examinations, the respiratory distress index (RDI) was assessed with overnight in-home polysomnography. Seated blood pressure (BP) was assessed at a baseline examination (t,) and after a 5-year follow-up period (t5), when 24-h ABPM also was performed. The change in RDI (t5-t1) over 5 years was significantly associated with 24-h mean systolic blood pressure (SBP) (P = .04), 24-h maximum diastolic blood pressure (DBP) (P = .03), sleep mean SBP (P = .05), sleep mean DBP (P < .05), and sleep maximum SBP (P = .02). Regression analysis revealed that average 24-h mean arterial pressure (MAP) and mean 24-h DBP were each best predicted by change in RDI, explaining 5% of the variance in these 24-h BP readings, and by current smoking status. After accounting for these variables, BP was not predicted by any of the other potential confounders (all P > .10). Mean RDI (averaged between t5 and t1) was associated with mean MAP, mean SBP, and maximal SBP measured during sleep. This study documents for the first time the association between changes in sleep apnea activity and BP and in a community based normotensive sample. Further long-term evaluation of the effects of these findings and the long-term consequences of hypertension are needed. PMID- 11497189 TI - Blunted nighttime blood pressure dipping in postmenopausal women. AB - Blunting of the normal drop in blood pressure (BP) from day to night is emerging as a strong prognostic indicator of cardiovascular morbidity and mortality. This study evaluated the effects of natural menopause on BP dipping in African American and white women. A total of 112 women (62 premenopausal and 50 postmenopausal) took part in the study. Pre- and postmenopausal groups were comparable in terms of clinic BP, body mass index, and ethnic composition. Ambulatory BP was recorded over 24 h during a typical workday, with measurements programmed to be taken every 15 min during waking hours and every 30-minutes during sleeping hours. Nocturnal BP dipping was defined as the difference between waking and sleep BP. Waking BP did not differ by menopausal status. However, nocturnal systolic BP (SBP) and diastolic BP (DBP) dipping were attenuated in postmenopausal women, with both SBP (P < .05) and DBP (P < .05) higher during nighttime sleep in postmenopausal than in premenopausal women. Ethnicity was also related to BP dipping, with African American women tending to show blunted SBP dipping (P = .055) compared with white women; BP dipping was most blunted in postmenopausal African American women. These observations suggest that blunted nighttime BP dipping may contribute to increased cardiovascular disease risk in postmenopausal women. PMID- 11497190 TI - Endothelin gene variants and aortic and cardiac structure in never-treated hypertensives. AB - BACKGROUND: The polymorphism of several candidate genes has been studied in relation to essential hypertension and cardiovascular complications. Target organ damage in essential hypertension is a complex disorder influenced by multiple genetic and environmental factors. The possible contribution of endothelin gene variants to target organ damage in hypertension in humans has not been studied in depth. PROCEDURE: We assessed the influence of genetic variants of components of the endothelin system ETAR -231A/G, 1363C/T, ETBR 30G/A and endothelin-1 (ET-1) 138insertion/deletion (I/D) on aortic stiffness, left ventricular geometric, and radial artery parameters in 528 never-treated hypertensive subjects of European origin. The study population included 314 men and 214 women with a mean age of 48+/-0.5 years (+/-SEM). In samples of patients, aortic stiffness was assessed with carotid-femoral pulse wave velocity (PWV). Radial artery thickness was measured with an echotracking angiometer and left ventricular geometric parameter with standard echographic procedures. RESULTS: The main results showed that the ETAR-231A/G (P = .022) and the ETBR 30G/A (P = .026) receptor gene variants influenced PWV level in women. The -231G and 30G alleles were associated with a codominant increase in PWV, explaining 18.6% of its variability (P = .005). In men, the ETBR 30G/A receptor gene variant was also related to the level of radial artery parameters (P = .02). No association between the 138I/D polymorphism of the ET-1 gene and left ventricular and radial artery parameters was observed in either men or women. CONCLUSIONS: These results indicate that the influence of endothelin system genes can be detected first on arterial parameters. PMID- 11497191 TI - Racial differences in resting end-tidal CO2 and circulating sodium pump inhibitor. AB - Previous studies have shown that high end-tidal CO2 (PetCO2) is a marker for sodium sensitivity of blood pressure (BP) in White Americans, and that the BP of African Americans is more sensitive to high sodium intake than that of whites. The present study tested the hypothesis that resting PetCO2 is higher in normotensive African Americans than in whites. Resting end-tidal CO2 of 395 white and 125 African American participants in the Baltimore Longitudinal Study on Aging was monitored for 20 min with a respiratory gas monitor, and BP and heart rate were recorded every 5 min by oscillometric methodology. Twenty-four-hour urinary excretion of a circulating sodium pump inhibitor marinobufagenin-like compound (MBG), which increases when plasma volume is expanded, was also analyzed by fluoroimmunoassay in racial groups. Mean resting PetCO2 of African American men was higher than that of white men (38.1+/-0.5 v 36.4+/-0.3 mm Hg), and resting PetCO2 of African American women was higher than that of white women (37.7+/-0.3 v 36.2+/-0.3 mm Hg). The differences were not significant in either men or women less than 50 years old, but were substantial in both men and women more than 50 years. Twenty-four-hour urinary excretion of MBG was higher in white (2.7+/-0.2 pmol) than in African American (2.1+/-0.2 pmol) participants, and high PetCO2 was a significant independent predictor of high MBG excretion in African Americans. These data are consistent with the hypothesis that the higher resting PetCO2 in African Americans plays a role in slower urinary excretion of sodium, greater BP sensitivity to high sodium intake, and increased prevalence of chronic hypertension. PMID- 11497192 TI - Similar effects of isolated systolic and combined hypertension on left ventricular geometry and function: the LIFE Study. AB - Echocardiograms of 143 patients with isolated systolic hypertension were compared to 808 patients with combined (systolic and diastolic) hypertension. All patients met electrocardiographic criteria for left ventricular hypertrophy and were evaluated off medication. Patients with isolated systolic hypertension were older, shorter, weighed less, and were mostly women, but body mass index (BMI) was similar in both groups. Systolic blood pressure (SBP) was 172 mm Hg in isolated systolic hypertension, 174 mm Hg in combined (P = not significant). Diastolic blood pressure was 83 and 101 mm Hg, respectively (P < .001). Despite having mean arterial pressure 12 mm Hg lower than patients with combined hypertension, the group with isolated systolic hypertension had equally severe abnormalities of left ventricular mass, left ventricular geometric patterns, and measures of systolic and diastolic function. Peripheral resistance was lower and pulse pressure/stroke volume ratio (arterial stiffness index) was higher and the isovolumic relaxation time shorter in isolated systolic hypertension. Multiple regression analyses identified age, height, BMI, stress-corrected mid wall shortening, stroke volume, male gender, and systolic or mean blood pressure (but not isolated systolic hypertension) as independent correlates of left ventricular mass. Relative wall thickness was independently associated with isolated systolic hypertension (P = .001) in addition to mean pressure and other covariates. The present results add support to the concept that systolic blood pressure (SBP) is a stronger determinant than diastolic pressure of cardiac target organ damage in hypertension. PMID- 11497193 TI - Relation of echocardiographic left ventricular mass and hypertrophy to persistent electrocardiographic left ventricular hypertrophy in hypertensive patients: the LIFE Study. AB - BACKGROUND: The Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) trial used left ventricular hypertrophy (LVH) on a screening ECG to identify patients at high risk for morbid events. Because of regression to the mean, not all patients who met screening criteria had persistent ECG LVH on the ECG performed at study baseline. METHODS: The relationship of echocardiographic LV mass and LVH to persistence or loss of ECG LVH between screening and baseline evaluation was examined in 906 hypertensive patients in the LIFE study, who had echocardiograms and additional ECG performed at study baseline. Patients were categorized according to the presence or absence of ECG LVH by Cornell voltage duration product criteria or Sokolow-Lyon voltage criteria; echocardiographic LVH was defined by LV mass index (LVMI) > 104 g/m2 in women and > 116 g/m2 in men. RESULTS: A total of 678 patients (75%) had persistent ECG LVH at baseline evaluation. Compared with the 228 patients without ECG LVH on the second ECG by either criterion, the 106 patients with LVH by both Cornell product and Sokolow Lyon criteria had significantly higher LVMI (140+/-31 v 114+/-21 g/m2, P < .001) and a higher prevalence of echocardiographic LVH (86% v 55%, P < .001). Patients with ECG LVH on the baseline ECG by either Cornell product criteria (n = 410) or Sokolow-Lyon voltage criteria (n = 162) had intermediate values of LVMI (125+/-25 and 121+/-21 g/m2) and prevalences of echocardiographic LVH (78% and 62%). After controlling for possible effects of age, sex, ethnicity, systolic blood pressure, and body mass index, persistence of ECG LVH on the baseline ECG was associated with an increased risk of echocardiographic LVH: compared with patients with neither ECG criteria for LVH, patients with only Sokolow-Lyon voltage criteria had a 1.2-fold increased risk of echocardiographic LVH, those with only Cornell product criteria had a 2.7-fold increased risk, and patients with both ECG criteria had a 4.1-fold increased risk of echocardiographic LVH (P < .001). CONCLUSIONS: Persistent ECG LVH between screening and LIFE study baseline identified patients with greater LV mass and a higher prevalence of echocardiographic LVH, suggesting that these patients may be at higher risk for subsequent morbid and mortal events. PMID- 11497194 TI - Screening for eligibility in the study of antihypertensive medication in children: experience from the Ziac Pediatric Hypertension Study. AB - BACKGROUND: The FDA Modernization Act has resulted in an increase in pediatric trials of antihypertensive medications. As experience is limited in children to guide the planning of these studies, we reviewed data from the Ziac Pediatric Hypertension Study to determine patterns of early study termination to help future studies. METHODS: For inclusion, subjects aged 6 to 17 years were required to have an average systolic blood pressure (SBP) or diastolic blood pressure (DBP) above the 95th percentile at the last of three visits during 2 weeks of single-blind placebo screening. Early study termination was defined as early termination for any reason. Screening termination was defined as normalization of blood pressure (BP) during the placebo screening phase. RESULTS: Early study termination rate was 27% (38 of 140 subjects). The most common reason was screening termination due to normalization of BP, accounting for 63% of all early study terminations. Among screening termination subjects who completed three screening visits, SBP was higher (P < .001) at visit 1 (129+/-8 mm Hg) than at visit 2 (123+/-7 mm Hg) or visit 3 (121+/-8 mm Hg), but did not differ between visits 2 and 3. Screening termination occurred in 15% with isolated SBP hypertension, and 21% with isolated DBP hypertension. At randomization, 83% had SBP hypertension and 53% had DBP hypertension. CONCLUSIONS: These data suggest that SBP hypertension should be part of inclusion criteria to increase enrollment and reduce the rate of screening termination, and that 1-week placebo screening is necessary and sufficient to minimize inclusion of transiently hypertensive subjects. PMID- 11497195 TI - Efficacy and safety of omapatrilat with hydrochlorothiazide for the treatment of hypertension in subjects nonresponsive to hydrochlorothiazide alone. AB - This multicenter, double-blind study evaluated efficacy and safety of the vasopeptidase inhibitor omapatrilat, which simultaneously inhibits neutral endopeptidase and angiotensin converting enzyme, when given in conjunction with hydrochlorothiazide (HCTZ) to subjects nonresponsive to HCTZ alone. The study enrolled 657 subjects with mild to severe hypertension. After a 2-week placebo lead-in period and a 4-week HCTZ phase, 274 subjects were randomized to receive omapatrilat (10 or 20 mg, electively titrated to 20 or 40 mg, respectively, at week 4 if seated diastolic blood pressure [SeDBP] was > or =90 mm Hg) or matching placebo in addition to 25 mg of HCTZ as continuing therapy. The primary outcome measure was change in SeDBP from baseline to week 8. At week 8, placebo plus HCTZ adjusted additional reductions in SeDBP in the omapatrilat 10/20 mg and 20/40 mg treatment groups (4 and 5 mm Hg, respectively) were significant (P < .001), as were changes in seated systolic blood pressure in both omapatrilat-treated groups (7 and 10 mm Hg, respectively; P < .001). Seated diastolic blood pressure was normalized (<90 mm Hg) in 38% of subjects in the placebo group compared to 59% and 64% of subjects in the omapatrilat groups (P < or = .008). Adverse events, serious adverse events, and discontinuations attributed to adverse events were infrequent. There were no clinically relevant changes in serum creatinine or potassium. Omapatrilat was effective and well tolerated when added to HCTZ in subjects whose blood pressure was not controlled with HCTZ alone. PMID- 11497196 TI - A study of the temporal relationship between plasma high molecular weight angiotensinogen and the development of pregnancy-induced hypertension. AB - In addition to the normally prevalent low molecular weight angiotensinogen (LMrA), significant quantities of a high molecular weight angiotensinogen (HMrA) are present in the human pregnant state. Previous studies have documented that 47% of women who develop pregnancy-induced hypertension (PIH) have a significantly elevated plasma HMrA/LMrA ratio. The purpose of this study is to establish whether or not the increase in the HMrA/LMrA ratio precedes the development of hypertension. Serial plasma samples were collected from a group of women throughout their pregnancy. High molecular weight angiotensinogen and LMrA levels in the samples from these women were determined. Fifteen of these women developed PIH. Seven women in the PIH group had a significantly elevated plasma HMrA/LMrA ratio. There was no consistent relationship between the elevation of the HMrA/LMrA ratio and the onset of hypertension. Three women had an elevated HMrA/LMrA ratio before the development of hypertension. In one woman the two events occurred simultaneously, and in three women the HMrA/LMrA ratio was elevated only after the development of hypertension. The current study shows that the development of hypertension during pregnancy is not the primary biologic signal for elevation of the plasma HMrA/LMrA ratio. Other parameters associated with fetal distress or abnormal development of placental circulatory systems must be involved in increasing the HMrA/LMrA ratio. It is proposed that the elevation of the HMrA/LMrA ratio is a mechanism by which the placental tissue specific renin-angiotensin system is attenuated. PMID- 11497197 TI - Association of increased pulse pressure with the development of heart failure in SHEP. Systolic Hypertension in the Elderly (SHEP) Cooperative Research Group. AB - The aim of this study was to assess the relationship between pulse pressure (PP) and the occurrence of heart failure (HF) in older persons with isolated systolic hypertension. Data from a prospective, multicenter, randomized, double-blind, placebo-controlled clinical trial were analyzed. A total of 4736 persons aged > or = 60 years with systolic blood pressure (SBP) between 160 and 219 mm Hg and diastolic blood pressure (DBP) < 90 mm Hg who participated in the Systolic Hypertension in the Elderly Program (SHEP) were studied. The main outcome measures were fatal and nonfatal HF. During 4.5 years average follow-up, fatal or nonfatal HF occurred in 160 of 4736 patients. The SBP, PP, and mean arterial pressure (MAP) were strong predictors of the development of HF (P < .0002). Cox proportional hazards regression using time-dependent covariates and controlling for MAP indicated that HF was inversely related to DBP (P = 0.002) and was directly related to pulse pressure (P = 0.002). Data were similar when patients who developed myocardial infarction during follow up were excluded. These data indicate that, in older persons with isolated systolic hypertension, high pulse pressure is associated with increased risk of heart failure independently of MAP and of the occurrence of acute myocardial infarction during follow-up. PMID- 11497198 TI - Effects of ACE inhibitor therapy on derived central arterial waveforms in hypertension. AB - Large artery properties constitute an important component of left ventricular afterload in hypertension. The present study examined whether such properties were particularly responsive to angiotensin converting enzyme inhibitor therapy. A prospective, randomized, 12-week study in 138 previously treated hypertensive subjects, in 67 of whom usual treatment (UC) was replaced with perindopril (P) therapy. Large artery properties were assessed as central arterial pressure augmentation determined by applanation tonometry of the radial artery and a transfer function. At baseline both augmentation index (AI, %) and pressure (AP, mm Hg) were related to body size, heart rate, and gender. In addition AP was related to age and systolic blood pressure (BP). After 12 weeks of treatment AP decreased significantly in both perindopril and UC groups, whereas AI only decreased significantly (151.7%+/-2.3% to 144.9%+/-2.6%) in those treated with perindopril. Decreases in AP (-4.2+/-0.9 mm Hg v -1.9+/-0.7 mm Hg) and AI ( 6.8%+/-2.2% v -2.2%+/-2.5%) from week 0 to week 12 were greater in the perindopril-treated group, but differences between groups failed to reach statistical significance (P = .05 and .09, respectively). The change in AI during the 12-week treatment period was dependent on the initial heart rate (P < .001), systolic BP (P < .05), weight (P < .001), and sex (P < .001), but not on treatment group (P > .5). Al at 12 weeks was negatively correlated with heart rate but regression slopes for the association were virtually identical for perindopril and UC groups. Perindopril treatment produces a greater decrease in AI than continuation with previous therapy, but this can be largely explained by hemodynamic changes rather than direct arterial effects. PMID- 11497199 TI - Long-term fructose feeding impairs vascular relaxation in rat mesenteric arteries. AB - To investigate the long-term influence of insulin resistance and hyperinsulinemia on vascular reactivity, both muscarinic and alpha2-receptor-mediated relaxations and the contribution of nitric oxide to these mechanisms were studied in the fructose-fed rat. Male Sprague-Dawley rats were fed either fructose-rich chow (FFR, n = 6) or normal chow (CNT, n = 6) for 40 weeks. Systolic blood pressure was measured by tail-cuff method. A 3-mm segment of mesenteric artery was excised, cannulated and pressurized, pretreated with prazosin (10(-6) mol/L) and propranolol (3 x 10(-6) mol/L), then precontracted with serotonin (10(-6) mol/L). Endothelium dependent relaxation was induced by addition of acetylcholine (10(-9) to 10(-4) mol/L), or a selective alpha2-agonist B-HT 920 (10(-9) to 10(-5) mol/L), with or without the nitric oxide synthase inhibitor L-NAME (10(-4) mol/L). Systolic blood pressure was significantly higher in FFR at the early period; however, there was no difference at the end of 40 weeks compared to CNT. Fasting plasma insulin was much higher in FFR than in CNT (110+/-62 v 41+/-11 microU/mL, P < .05), whereas plasma glucose was not different. Maximum relaxation to acetylcholine was attained at 10(-6) mol/L in FFR but at 3 x 10(-7) mol/L in CNT. The degree of maximum relaxation attained with acetylcholine was similar in FFR and CNT (89+/-9 and 94+/-4% of precontraction), although attenuated (P < .01) by the addition of L-NAME only in FFR (to 34+/-22%, P < .05) but not in CNT (to 82+/-25%). The half-maximal relaxation dose of acetylcholine was greater in FFR (P < .01) compared with CNT and was significantly increased (P < .05) by L-NAME in both groups. B-HT 920 at 10(-5) mol/L induced a greater relaxation in CNT (36+/-10% of serotonin constriction) than in FFR (19+/-14%, P < .05). These responses were significantly blunted by L-NAME. Thus, muscarinic receptor mediated vascular relaxation is less sensitive and more nitric oxide dependent in FFR versus CNT. Alpha2-adrenergic-mediated relaxation, predominantly mediated by nitric oxide, is also impaired in FFR. It is possible that prolonged insulin resistance and hyperinsulinemia in FFR could alter endothelial-dependent vasodilatory mechanisms, thereby contributing to the increase in blood pressure seen in this model. PMID- 11497200 TI - Silent cerebral infarcts in basal ganglia are advanced in congenital protein C deficient heterozygotes with hypertension. AB - Congenital protein C deficiency is now widely recognized as a genetic risk for venous thrombosis. However, it remains uncertain whether this condition also confers risk for arterial thrombosis. We evaluated the association of congenital protein C deficiency with hypertension and silent cerebrovascular disease using brain magnetic resonance imaging (MRI) (T1- and T2-weighted and proton density images) in a large family pedigree of protein C deficiency diagnosed by gene analysis, compared with 46 non-pedigree related control subjects with normal protein C levels (> or = 75%) who were selected from among 55 asymptomatic hypertensive subjects matched for age and cardiovascular risk factors. Of the 58 living subjects in this pedigree, we measured plasma protein C levels in 45 subjects, and found 2 cerebral infarctions in the 24 heterozygotic subjects, whereas there was no stroke in the 21 normal homozygotic subjects. We performed brain MRI in 14 asymptomatic hypertensive subjects without any cardiovascular disease and in two patients with cerebral infarction, and found 28 cerebral infarcts (two corresponded to the patients' neurologic deficits and 26 were silent). All were lacunar infarcts < 10 cm in size. A total of 25 silent lacunar infarcts were found in nine heterozygotic subjects, whereas only one was found in the seven normal homozygotic subjects (2.8 v 0.14 lacunes per person, P = .002). No advanced white matter hyperintense lesions in T2-weighted images were found in either group. The prevalence of silent lacunar infarcts in the heterozygotic subjects was also significantly higher than that in normal control subjects (1.0 per person, P = .01). Concerning the distribution of silent infarcts, the number of lacunes located in the basal ganglia was higher in the heterozygotic subjects (2.3 per person, P < .001) than in the seven normal homozygotic subjects (0.14 per person) or in the control group (0.28 per person), whereas the number of lacunes in the white matter was not different among the groups. In conclusion, congenital protein C deficiency may accelerate the progression of silent cerebral infarct formation in hypertension, particularly in the basal ganglia, and may be a potential risk for stroke or vascularly induced dementia. PMID- 11497201 TI - A cross-sectional, diurnal, and follow-up study of platelet activation and endothelial dysfunction in malignant phase hypertension. AB - To investigate the hypothesis that abnormalities of thrombogenesis and endothelial damage/dysfunction are greater in malignant hypertension (MHT) compared with uncomplicated nonmalignant essential hypertension (EHT) > 160/90 mm Hg), we measured markers of endothelial function (von Willebrand factor) platelet activation (soluble P-selectin) and fibrinogen in 18 consecutive patients with MHT, 50 patients with untreated EHT, and 34 healthy control subjects. We also investigated whether there was any diurnal variation in the measured indices, as well as the effects of good blood pressure (BP) control after 6-month follow-up. Mean plasma fibrinogen and von Willebrand factor levels were both highest in the MHT group, intermediate in the nonmalignant hypertension group and lowest in the normotensive control subjects (P < .001). Similarly, mean soluble P-selectin levels were higher in both hypertensive groups compared to normotensive control subjects (P = .033). There was no significant diurnal variation in plasma fibrinogen, soluble P-selectin, and von Willebrand factor levels over the 24-h study period among the MHT patients. At 6-month follow-up and a reduction in mean BP, there was no significant change in mean plasma fibrinogen levels (P = .25), but both soluble P-selectin (P < .001) and von Willebrand factor (P = .0025) were significantly reduced. In conclusion, malignant hypertension is associated with abnormal endothelial damage (elevated von Willebrand factor), platelet activation (soluble P-selectin), and fibrinogen levels, which may be related to the pathogenesis of this condition, as well as the development of complications. These abnormalities do not undergo any significant diurnal variation and may be beneficially altered by BP reduction. PMID- 11497202 TI - Severe and refractory hypertension in a young man. PMID- 11497203 TI - Treatment of atheroembolization with corticosteroids. AB - Aortic atheroembolization is a feared complication of invasive procedures such as arteriography, often leading to devastating complications including renal insufficiency. To date, even in cases with evolving renal failure, there is no recommended treatment. This case report describes the successful treatment with corticosteroids of a patient with deteriorating renal function after renal arteriography and angioplasty, resulting in rapid and sustained improvement in renal function. The implications of this observation are discussed. PMID- 11497204 TI - Effect of indomethacin on blood pressure in elderly people with essential hypertension well controlled on amlodipine or enalapril. PMID- 11497205 TI - Beyond the sphygmomanometric numbers: hypertension as a syndrome. AB - Blood pressure is a critical element of hypertension, but evidence shows that many other interrelated risk factors--such as hypercholesterolemia, left ventricular hypertrophy, insulin resistance, and vascular dysfunction--contribute to form a complex syndrome of hypertension. Studies have demonstrated that these other risk factors often precede the onset of elevated blood pressure (BP). Significantly, this has been shown in the normotensive offspring of hypertensive parents. One factor that the elements of the hypertension syndrome have in common is endothelial dysfunction. An upset in the normal balance of angiotensin II and nitric oxide in the endothelial cell is associated with manifestation of the hypertension syndrome-associated risk factors. Recognition of this fact can and should be constructively incorporated into clinical practice in three ways: earlier identification of patients with the hypertension syndrome, earlier treatment to prevent disease onset/progression, and the use of angiotensin converting enzyme inhibitors to correct the critical imbalance in the endothelial cell. PMID- 11497206 TI - Arterial compliance to stratify cardiovascular risk: more precision in therapeutic decision making. AB - The focus of attention in preventing and treating cardiovascular (CV) disease today is shifting toward the arterial wall. Evidence has been accumulating for several years that protecting the endothelium is key to reducing CV risk. Endothelial dysfunction results in reduced compliance, or increased arterial stiffness, particularly in the smaller arteries. This abnormality is characteristic of patients with hypertension but may also be seen in normotensive patients before the appearance of clinical disease. Reduced arterial compliance is also seen in patients with diabetes and in smokers, and is part of a vicious cycle that further elevates blood pressure, aggravates atherosclerosis, and leads to increased CV risk. Although other factors are involved, the damage to the endothelium results in reduced secretion of nitric oxide, which influences smooth muscle growth, migration, and contraction, as well as influencing inflammation and clotting. Arterial compliance can be measured by several techniques, most of which are invasive or otherwise not clinically appropriate. Pulse contour analysis is a newly developed noninvasive method that allows for easy, in-office measurement of arterial elasticity to identify patients at risk for CV events before disease becomes clinically apparent. Further research is needed to confirm whether this method offers a means of improving risk stratification and therapeutic decision making. PMID- 11497207 TI - Angiotensin-converting enzyme inhibition to enhance vascular health--clinical and research models. AB - Protection of the endothelium, the metabolically active inner lining of the vasculature, appears to be a key factor in maintaining cardiovascular (CV) health. The endothelium responds to hemodynamic and hormonal factors by secreting substances that maintain vascular homeostasis. Damage to the endothelium is an initial step in the development of CV disease. Angiotensin-converting enzyme (ACE) inhibitors, which block the formation of the vasoconstricting substance, angiotensin II, have proved to be a key therapy for hypertension and congestive heart failure. The activity of these agents in enhancing vascular health appears to be a critical factor in their therapeutic effectiveness. Large-scale clinical trials over the past decade have shown that ACE inhibition is an effective therapeutic means of not only prolonging survival and reducing morbidity after acute myocardial infarction, but also reducing all-cause mortality and morbidity in patients at high risk for CV disease, including patients with diabetes. ACE is found in far greater amounts in tissue than in plasma. Studies indicate that ACE inhibitors act at the tissue level to provide long-term cardioprotective effects that include a reduction in the progression of atherosclerosis. An issue to resolve is how much ACE inhibition is needed at the tissue level to reverse or prevent further vascular damage. PMID- 11497208 TI - Ongoing trials of angiotensin-converting enzyme inhibition: what they can tell us. AB - Abundant evidence has accumulated showing that angiotensin converting enzyme (ACE) inhibitors reduce long-term cardiovascular morbidity and mortality rates in patients with heart failure and myocardial infarction. Fewer completed trials have assessed their potential benefits in this regard in hypertensive subjects, but evidence of benefit is beginning to accrue from studies examining patients with hypertension, particularly in the presence of diabetes and after infarction. Ongoing trials of blood pressure (BP) lowering using ACE inhibition fall into three main categories: 1) those comparing ACE inhibitors with older drugs such as diuretics and beta blockers; 2) those examining more aggressive versus less aggressive lowering of BP; and 3) those investigating BP lowering in patients at high risk for a cardiac event. Among those in the last group is the Perindopril Protection Against Recurrent Stroke Study (PROGRESS), which examines the effects of perindopril-based ACE inhibitor therapy in both normotensive and hypertensive patients who have survived a stroke. This trial is particularly important because it serves as a model for studies of BP lowering across a wide range of BP and BP related conditions. PMID- 11497209 TI - The nature of human orgasm: a critical review of major trends. AB - This critical review presents a synthesis of the available theoretical and empirical literatures on human orgasm. Findings from both normal and clinical human populations are included. Two major trends in the literature, the dichotomization of biological and psychological perspectives and the assumption of gender differences, are highlighted. A new multidimensional model of the psychological experience of orgasm is described with a view to futhering a biopsychological approach applicable to both sexes. Clinical applications of this new model are discussed. PMID- 11497210 TI - Chronic pain and posttraumatic stress disorder: mutual maintenance? AB - Common sequelae following a traumatic event include chronic pain and posttraumatic stress disorder (PTSD). Over the last decade, the literature relating to PTSD has become progressively more sophisticated, resulting in well supported theories and treatments for sufferers. Equivalent research relating to chronic pain has more recently gathered momentum. However, to date there has been minimal attention devoted to the concurrence of the two disorders, even though high comorbidity has been noted. This review begins by briefly summarizing the literature relating to the two disorders in terms of symptoms, prevalence and comorbidity. It explicates the major psychological theories of chronic pain and PTSD and reviews the evidence relating what factors maintain the disorders. A number of pathways by which chronic pain and PTSD may be mutually maintaining are highlighted. We conclude that chronic pain and PTSD are mutually maintaining conditions and that there are several pathways by which both disorders may be involved in the escalation of symptoms and distress following trauma. Treatment implications are considered, as are issues for future research. PMID- 11497211 TI - Perfectionism and psychopathology: a review of research and treatment. AB - Clinical experience suggests that perfectionism can impede the successful treatment of psychological disorders. This review examines the concept of perfectionism, critically evaluates its assessment, reviews the association between existing measures of perfectionism and psychopathology, and considers the impact of perfectionism on treatment. It is concluded that existing measures do not reflect the original construct of perfectionism and that, consequently, new measures are needed. The evidence reviewed indicates that high personal standards are specifically elevated in patients with eating disorders and beliefs about others' high standards for the self are associated with a broad range of psychopathology. The importance of examining mean scares across studies (as well as associations between variables within studies) is emphasized. There has been no systematic evaluation of the treatment of perfectionism despite existing cognitive-behavioral treatment protocols. PMID- 11497212 TI - Biological contributions to the presentation and understanding of attention deficit/hyperactivity disorder: a review. AB - Attention-deficit/hyperactivity disorder (ADHD) is the most commonly diagnosed disorder in children today with estimated prevalence rates falling between 3 and 5% of children (American Psychiatric Association, 1994). From inception, research has focused on studying varying facets of this disorder with initial efforts primarily focusing on treatment outcome. However, prominent efforts have been made in recent research efforts to shed light on the etiology of this disorder. Such research has discovered the contribution of genetic inheritance, as well as environmental factors that lead to the development of this disorder. Furthermore, studies using neurological and neuropsychological assessment measures have implicated the involvement of various Parts of the brain. This article critically reviews this body of research in light of its impact on the current specific neuropsychologically based etiological theories, as well as the most beneficial directions for future research. PMID- 11497213 TI - Posttraumatic stress disorder and traumatic brain injury: can they co-exist? AB - The possibility that posttraumatic stress disorder (PTSD) can develop following traumatic brain injury (TBI) has been the subject of considerable debate. The traditional view has held that impaired consciousness that occurs with TBI precludes encoding of the traumatic experience, and this prevents subsequent reexperiencing symptoms. This paper critically reviews available, empirical studies on PTSD in TBI populations and suggests that these two conditions can co exist. The various mechanisms that may mediate PTSD following TBI are discussed, and special attention is given to issues that recognize the distinctive features of PTSD following TBI. These processes include implicit processing, biologically mediated fear conditioning, and reconstruction of trauma memories. Finally implications for assessment, treatment, and forensic investigation of PTSD in TBI populations are, addressed. This review concludes that TBI populations provide a useful means by which the role of traumatic memories (and impaired memories) in posttraumatic adjustment can be studied. PMID- 11497214 TI - Body dysmorphic disorder: a review of conceptualizations, assessment, and treatment strategies. AB - Increased interest in body dysmorphic disorder (BDD) has generated a wealth of recent research. This paper reviews the current literature regarding conceptualizations of the disorder, the development of assessment tools, and treatment outcome. Although BDD has been viewed as a variant of an eating disorder, obsessive compulsive disorder, or a somatoform disorder, it appears best conceptualized as a body image disorder with social, psychological, and possibly biological influences. Assessment instruments with acceptable psychometric properies have been developed to specifically assess BDD (e.g., the Body Dysmorphic Disorder Examination and the Yale-Brown Obsessive Compulsive Scale modified for Body Dysmorphic Disorder). Examination and the initial results from uncontrolled and controlled treatment research suggest that cognitive behavioral treatments for BDD may be as effective as those for possibly related disorders such as obsessive compulsive disorder and bulimia nervosa. PMID- 11497215 TI - Prevention of bacterial diarrhea by pasteurization of drinking water in Kenya. AB - Diarrheal disease is one of the major causes of morbidity and mortality in developing countries. Drinking water is a primary transmission route of infectious diarrheagenic bacteria in a rural area of Kenya (Microbiol. Immunol. 41: 773-778, 1997). We tried to prevent diarrhea at villages with approximately 1,500 households in Kenya by pasteurizing drinking water. A durable simple thermoindicator which changes color at 70 C was used as an indicator of pasteurization. The number of households in which drinking water was coliform bacteria-free increased from 10.7% to 43.1% after adoption of a pasteurization practice. Consequently, the incidence of severe diarrhea among people drinking pasteurized water was significantly lower than in people taking raw water (odds ratio=0.55, P=0.0016). The reduction ratio of the incidence after pasteurization was nearly equivalent with that after the adoption of a boiling method. Employment of women leaders as fieldworkers and demonstration of bacterial colony disappearance on agar plates by pasteurization also affected reduction of the diarrheal incidence. PMID- 11497216 TI - Gene analysis of Vibrio cholerae NAGV14 pilus and its distribution. AB - Adhesive pilus of Vibrio cholerae 034, strain NAGV14, was genetically analyzed. The deduced amino acid (aa) sequence of the major pilin structural gene (VcfA) was 67% homologous to the MshA pilin in the N-terminal region, but no homology was found in the C-terminal region which contained the antigenic epitopes. Upstream and downstream flanking regions examined were highly homologous to mshB and mshC of the MSHA (mannose-sensitive hemagglutinin) gene locus. A short leader sequence and a pair of cysteines near the C-terminus which are the characteristics of type 4a pilus family were found. The major pilin structural gene of NAGV14 was compared to that of a strain V10 producing non-adhesive pili. The deduced aa sequences showed 60% homology, and the distance between two cysteines in the C-terminal region was different. A total of 177 V. cholerae strains were investigated for the presence of a type 4 pilus gene locus by PCR, and 95% were positive. The major pilin gene of NAGV14 was detected in 4 of 93 V. cholerae non-O1, non-0139 strains tested, but none of the V. cholerae O1 and O139 (72 and 12 strains, respectively). Our result suggested that a type 4 pilus gene locus similar to the MSHA gene locus is widely distributed among V. cholerae strains. We proposed naming this type 4 pilus gene locus the VCF (for V. cholerae flexible pili) gene locus. PMID- 11497217 TI - Protective effect of OK-432 on mice against endotoxemia and infection with Pseudomonas aeruginosa and Salmonella enteritidis. AB - OK-432 has been used clinically as a biological response modifier for cancer therapy. We investigated here the protective effects of OK-432 against endotoxic shock and infectious death caused by Pseudomonas aeruginosa and Salmonella enteritidis in mice and proposed a possible mechanism. Pretreatment of OK-432 reduced the lethality of lipopolysaccharide (LPS)-induced endotoxic shock in D (+)-galactosamine-sensitized C3H/HeN mice. OK-432 did not affect the TNFalpha production in blood, but it did decrease the susceptibility to TNFalpha. Furthermore, an acceleration of LPS clearance from blood was detected. The pretreatment of OK-432 also decreased the lethality of mice in bacterial infection caused by P. aeruginosa and S. enteritidis. The rapid decrease of the viable bacteria from the circulating blood and in spleen and liver in mice was observed in a manner similar to LPS clearance. These findings indicate that the protective effect of OK-432 against the endotoxemia and bacteremia may depend on an up-regulation of clearance of LPS and bacteria and the augmented resistance to TNFalpha. PMID- 11497218 TI - Molecular heterogeneity of heat-stable toxin-producing and non-producing Escherichia coli O25 strains isolated in Japan. AB - Escherichia coli O25 strains that produce heat-stable toxin (ST) have been recently isolated in Japan, and epidemiological study of this type of enterotoxigenic E. coli is required. In this study the heterogeneity of 16 ST producing and non-producing strains of E. coli O25 was investigated. All eight ST producing strains were shown to have STIb gene, and seven of them had similar profiles of plasmids, ladder-banding of LPS in SDS-polyacrylamide gel electrophoresis, and chromosomal DNA digestions in pulsed-field gel electrophoresis (PFGE). In contrast, ST-non-producing strains were more heterogeneous in all parameters examined. PFGE of the digested chromosomal DNA with several restriction enzymes was proved to be an effective procedure to compare the closely related strains of E. coli O25. PMID- 11497219 TI - Epidemiological survey of Orientia tsutsugamushi distribution in field rodents in Saitama Prefecture, Japan, and discovery of a new type. AB - There are various antigenic variants of Orientia tsutsugamushi which are distinguished by immunological and molecular genetic methods targeted at the antigenic diversity of 56-kDa type-specific antigen proteins. The present study was performed to analyze 15 strains successfully isolated from rodents in Saitama Prefecture, Japan, by 56-kDa gene sequence homologies, reactivities with type specific monoclonal antibodies and polymerase chain reaction (PCR) using type specific primer-pairs. We demonstrated the presence of a new type of O. tsutsugamushi among the isolates. This new type, designated as the Saitama type, was located in the branch of Karp type in the phylogenetic tree based on 56-kDa gene sequences, but distant from the known Karp types, such as Karp, JP-1 and JP 2, showing less than 90% homology. Strains of this type could not be distinguished by immunological methods from Karp type strains, but a new primer pair for PCR which specifically amplifies the DNA of this new type strain was designed. This primer-pair may serve to find this strain type in future studies. PMID- 11497220 TI - Growth inhibition of cancer cells by co-transfection of diphtheria toxin A-chain gene plasmid with bovine leukemia virus-tax expression vector. AB - We constructed a plasmid containing bovine leukemia virus (BLV)-tax gene driven by SR alpha promoter, designated as pME-BLVtax, to activate the promoter of the long terminal repeat (LTR) of BLV in various tumor cells. Activation of the promoter of BLV-LTR by pME-BLVtax was confirmed by luciferase assay. When the cells, such as COS-1, C8, and KU-1, were transfected with a plasmid pBLV-LUC1, which contained the luciferase gene under the control of BLV-LTR, and pME-BLVtax, luciferase was expressed in these cells, whereas no luciferase gene expression was observed when only pBLV-LUC1 was introduced into the cells. Activation of the BLV-LTR promoter was regulated by pME-BLVtax and 0.5 microg of pME-BLVtax was sufficient for the expression of the gene under the control of BLV-LTR. Furthermore, pME-BLVtax was used to direct the cell expression of the gene for diphtheria toxin A-chain under the control of BLV-LTR (pLTR-DT) to various tumor cell lines, KU-1, C8, COS-1, BL2M3, and HeLa cells. The transfection was carried out with cationic liposomes. In this experiment, co-transfection of pLTR-DT with pME-BLVtax exerted selective growth inhibitory effects on the tumor cell lines. Moreover, three co-introductions of pLTR-DT with pME-BLVtax into the cell lines resulted in significant inhibition of the cell growth. This result suggests that the delivery of the pLTR-DT and pME-BLVtax genes into tumor cells by the use of cationic liposomes may be potentially useful as a novel approach for the treatment of tumor cells. PMID- 11497221 TI - (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assays showed a lack of toxicity of ganciclovir (GCV), C.OXTs, and their derivatives, to TaY(OK) cells at 1 microM. Therefore we compared the antiviral potencies of these drugs at 1 microM by monitoring the viral loads produced during a 1-day period during the course of the drug treatment. Among the drugs tested, 3'-fluorocarbocyclic oxetanocin A (3'F-C.OXT-A) was the most effective for inhibiting the virus production, and at concentrations ranging from 0.5 microM to 10 microM, the inhibition of the viral production was dose-dependent. A comparison of the chemical structures of the derivatives with that of C.OXT-A, which is the parental molecule, suggested that the 3'-fluorine-modification might account for the higher anti-HHV-6 activity and lower cytotoxicity. AB - An infection of TaY cells, which originated from an adult T-cell leukemia, with an HHV-6B OK isolate resulted in a chronically infected culture, termed TaY(OK). Cell cloning analysis revealed that the TaY(OK) culture consisted of a mixture of cells permissive and refractory to the infection, and that the permissive cells were continuously produced from the refractory cell population. Since the chronically infected culture has been maintained for over 2 years without the addition of uninfected TaY cells, we used it for an evaluation of the antiviral potency of nucleoside analogs, especially carbocyclic oxetanocins (C.OXTs). PMID- 11497222 TI - A sandwich ELISA for detection of soluble GPI-80, a glycosylphosphatidyl-inositol (GPI)-anchored protein on human leukocytes involved in regulation of neutrophil adherence and migration--its release from activated neutrophils and presence in synovial fluid of rheumatoid arthritis patients. AB - GPI-80, mainly distributed on human neutrophils and monocytes, is a novel glycosylphosphatidylinositol (GPI)-anchored protein regulating neutrophil adherence and migration. We prepared a polyclonal antibody specific to GPI-80 by immunizing rabbits with the purified GPI-80 and established a sandwich ELISA for detecting soluble GPI-80. We found that soluble GPI-80 was released from fMLP activated neutrophils and was present at high concentrations in synovial fluids but not sera of rheumatoid arthritis patients, suggesting that GPI-80 may play a role in inflammatory diseases. The detection of soluble GPI-80 may help us to understand its physiological and pathological functions. PMID- 11497223 TI - Suppressive mechanisms of EPA on human T cell proliferation. AB - In vivo and in vitro experiments show that polyunsaturated fatty acids (PUFAs) including eicosapentaenoic acid (EPA) inhibit mitogen- or antigen-stimulated proliferation of T cells in rodents and humans. However, the exact manner and mechanisms by which PUFA inhibits T cell proliferation is not clear. In the present study, we investigated the suppressive effects of EPA, an n-3 PUFA, on PHA stimulated human peripheral blood T cells. Our results showed that EPA suppresses mitogen- or antigen-stimulated human T cell proliferation by at least 2 steps; step 1) EPA suppresses T cell proliferation by inhibiting IL-2R alpha expression and IL-2 production; step 2) EPA induces cell death of blast T cells without reducing the expression of IL-2R alpha. We also showed that EPA selectively stimulates the cell death of blast T cells but not resting T cells. The suppressive effect of EPA was mediated via the production of reactive oxygen products, because EPA-stimulated H2O2 production and the suppressive effect of EPA was restored by addition of catalase or NAC. These results taken together suggest that such immunosuppressive effects of EPA may explain the apparent benefits of EPA-enriched diets for patients with inflammatory disorders. PMID- 11497224 TI - Regulation of Fas-mediated apoptosis by NF-kappaB activity in human hepatocyte derived cell lines. AB - Nuclear Factor-kappaB (NF-kappaB) is known to be one of the key regulators of genes involved in survival signaling. The purpose of this study is to investigate the role of NF-kappaB activity in signaling events related to cell survival in hepatocytes, which has been supposed to be one of the most sensitive organs against Fas-induced cytotoxicity. The functions of NF-kappaB activity on Fas mediated apoptosis in different human cell lines were investigated by a magnetic concentration system for cells with exogenous protein production. We demonstrated that the activation of NF-kappaB was triggered by anti-Fas treatment in hepatocyte derived cell lines, HepG2 and Huh-7 cells. Overexpression of kinase inactive NF-kappaB-inducing kinase (NIK) and IkappaB kinase (IKK) inhibited the activation of NF-kappaB introduced by anti-Fas treatment in these cells. Notably, inactivation of NF-kappaB by the production of IkappaB-alpha protein made these cells more susceptible to apoptosis induced by Fas stimulation. In contrast, transient expression of NIK showed a suppressive effect on Fas-mediated apoptosis in the same cell lines. These findings suggest the involvement of NF-kappaB in suppression of Fas-mediated apoptosis in human hepatocyte derived cell lines, in which concomitant activation of this transcriptional factor was observed through the stimulation of Fas signaling. PMID- 11497225 TI - flaB-polymerase chain reaction (flaB-PCR) and its restriction fragment length polymorphism (RFLP) analysis are an efficient tool for detection and identification of Leptospira spp. AB - For establishment of a rapid-identification method of Leptospira species, a flaB gene of Leptospira was investigated and the following results were obtained. 1) HaeIII- or HindIII-restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR) products (793 bp) of flaB gene was effectual for the classification of species of Leptospira. 2) Twenty cells of Leptospira in 1 ml of coagulated blood and 100 cells of Leptospira in 1 ml of anti-coagulated blood could be detected by flaB-PCR. These results suggested that PCR-RFLP based on the flaB gene was an efficient tool for rapid detection and identification of species of infected Leptospira from clinical specimens. PMID- 11497226 TI - Reconstitution and purification of cytolethal distending toxin of Actinobacillus actinomycetemcomitans. AB - Cytolethal distending toxin (CDT) has been found in various pathogenic bacterial species and causes a cell distending and a G2 arrest against eukaryotic cells. All the cdtABC genes, which encode CDT, are known to be required for the CDT activities although the CDT holotoxin structure has not been elucidated. We cloned the cdtABC genes of Actinobacillus actinomycetemcomitans and constructed an Escherichia coli expression system for them. We found that crude extracts from six deletion mutants (delta cdtA, delta cdtB, delta cdtC, delta cdtBC, delta cdtAC, and delta cdtAB) of recombinant E. coli, which showed very weak or no detectable CDT activities, restored the CDT activities when pre-mixing and pre incubation of them were performed in combinations to contain all the CdtA, CdtB, and CdtC proteins. These results indicate that all the Cdt proteins are required for the CDT activities. We also found that the chimera CdtB protein, CdtB-intein CBD (chitin binding domain) like CdtB protein itself assembled with CdtA and CdtC. The reconstituted CDT containing the chimera CdtB protein was specifically extracted by chitin beads and the only CDT portion was isolated from the chitin beads by a cleavage reaction of the intein. The purified reconstituted-CDT was found to consist of CdtA, CdtB, and CdtC proteins, and showed appreciable CDT activities, indicating that the CDT holotoxin structure is the CdtABC complex. To our knowledge, this is the first report succeeded in complete purification of an active CDT and may offer useful tools for elucidation of the toxic mechanism of CDT. PMID- 11497227 TI - The emerging world role of irinotecan in lung cancer. AB - Irinotecan (CPT-11, Camptosar), either alone or in combination with cisplatin (Platinol), has demonstrated activity in advanced non-small-cell lung cancer (NSCLC). In single-agent studies, response rates as high as 35% have been observed; in combination with cisplatin, response rates have ranged as high as 50%, with 1-year survival rates of 33% to 58%. A critical phase III randomized trial comparing irinotecan, either alone or in combination with cisplatin, to vindesine/cisplatin, demonstrated superior survival for stage IV patients receiving irinotecan. The first North American effort to replicate the schedule used in the phase III trial (cisplatin 80 mg/m2 and irinotecan 60 mg/m2 on days 1, 8, and 15 every month) yielded a response rate of 29%, median survival time of nearly 10 months, and 1-year survival rate of 37%. A subsequent multi institutional trial conducted through Vanderbilt Cancer Center Affiliate Network and Fox Chase Cancer Center combined both agents on a weekly schedule in an attempt to exploit their putative synergy and to potentially decrease toxicity. This schedule, which employed irinotecan 65 mg/m2 and cisplatin 30 mg/m2 both weekly x 4, was better tolerated than the monthly cisplatin combination with a higher response rate (36%), median survival (11.6 months), and 1-year survival rate (46%). Multiple phase I and phase II studies have combined irinotecan with taxanes, either alone or in concert with carboplatin (Paraplatin), yielding similar response and survival rates. Finally, a critical phase III trial from Japan has demonstrated superior outcome for irinotecan and cisplatin vs standard etoposide/cisplatin in the treatment of extensive small-cell carcinoma of the lung. At least one North American trial will determine if these results are reproducible. PMID- 11497228 TI - Irinotecan and radiation in combined-modality therapy for solid tumors. AB - Irinotecan (CPT-11, Camptosar) is a camptothecin derivative that is thought to exert its cytotoxic effects by targeting topoisomerase I. It is believed that irinotecan stabilizes a DNA-topoisomerase I cleavable complex, and that interactions between this complex and the replication machinery may lead to cell death. There is a significant volume of in vitro and in vivo data demonstrating that irinotecan acts as a radiosensitizer. The exact mechanism of this radiosensitization is currently unknown. The increasing amount of data demonstrating improved outcomes with concurrent chemoradiation treatment of malignancies like lung cancer and head and neck cancer provide impetus for pursuing the addition of other drugs as radiosensitizers to improve local control further. Irinotecan is undergoing early clinical trials in the combined-modality setting in several disease sites. This article will provide an overview of the current status of irinotecan used concurrently with radiotherapy in the treatment of a variety of solid tumors. PMID- 11497230 TI - Recombinant human thrombopoietin in myelosuppressive chemotherapy. AB - Recombinant human thrombopoietin (rhTPO) is a full-length glycosylated molecule that has been under evaluation in the setting of chemotherapy-induced myelosuppression. It has been shown to be a potent stimulator of platelet production in cancer patients when administered prior to chemotherapy. The peak platelet response to a single dose of rhTPO is observed around day 12, and is accompanied by a significant increase in the number of mature megakaryocytes in bone marrow. Consistent with this biologic effect, rhTPO administered postchemotherapy has been shown to be effective in attenuating severe thrombocytopenia induced by carboplatin, which produces a late platelet nadir. Early clinical experience with a regimen that produces an early nadir, however, such as AI (doxorubicin [Adriamycin] and ifosfamide [Ifex]), suggests that administration of rhTPO both prior to and following chemotherapy might be important to reduce thrombocytopenia severity. Treatment with rhTPO in these clinical trials has been well tolerated with a favorable safety profile. Randomized clinical trials have been initiated to determine further the importance of schedule in the prevention and treatment of severe thrombocytopenia in cancer patients. PMID- 11497229 TI - Rationale for non-platinum chemotherapy in advanced NSCLC. AB - During the past decade, five new cytotoxic drugs have been introduced that are active against non-small-cell lung cancer (NSCLC). These agents include vinorelbine (Navelbine), paclitaxel (Taxol), docetaxel (Taxotere), gemcitabine (Gemzar), and irinotecan (CPT-11, Camptosar). Used alone, these drugs display activity comparable to cisplatin. The combination of cisplatin and one of the newer drugs produces better survival than treatment with cisplatin (Platinol) alone. Randomized studies of chemotherapy regimens that include these newer drugs have demonstrated improved survival, fewer side effects, or both, compared with earlier standard combinations such as cisplatin/vindesine or cisplatin/etoposide. Docetaxel and perhaps some of the other newer drugs are of value for patients previously treated with platinum-containing regimens. Future studies should determine whether combinations of these newer drugs are superior to cisplatin containing regimens. Although improved survival is the most important factor in defining the best regimen in non-small-cell lung cancer, additional considerations include patient tolerability, costs of administration, and the rationale for and ability to include noncytotoxic agents (such as inhibitors of signal transduction pathwriys or angiogenesis) into the therapeutic program. PMID- 11497231 TI - Role of angiogenesis inhibitors in cancer treatment. AB - Angiogenesis is essential for the growth of both primary and metastatic tumors. This process, more complex than was previously thought, requires the coordinated activities of multiple factors and cell types. For tumors to develop a neovascular blood supply, tumor and host cells must secrete pro-angiogenic factors that offset the activities of inhibitory angiogenic factors. In addition, the newly derived tumor endothelium must respond to survive in a relatively caustic microenvironment. Thus, endothelial-cell survival factors are essential in the maintenance of this neovasculature. Because redundant factors and pathways regulate angiogenesis, inhibition of any single pathway is unlikely to lead to prolonged response in most patients with solid malignancies. Since anti angiogenic therapy is unlikely to induce tumor regression, the criteria for efficacy must be evaluated by means other than the standard criteria used to evaluate cytotoxic chemotherapy regimens. Understanding the basic principles that drive tumor angiogenesis will lead to the development of therapies that will likely prolong survival without the toxicity associated with standard chemotherapy. PMID- 11497232 TI - Topoisomerase I inhibitors in the treatment of head and neck cancer. AB - Traditionally, the role of chemotherapy in the treatment of squamous carcinoma of the head and neck has been confined to patients with recurrent or metastatic disease who are deemed incurable with surgery or radiation therapy. Over the past decade, however, the role of chemotherapy has changed dramatically. The use of primary combined chemoradiation to preserve function or to enhance survival in patients with unresectable disease has become a standard approach. As the use of chemotherapy in squamous carcinoma of the head and neck has expanded, investigators have been interested in identifying new active agents. Topoisomerase I inhibitors, a new class of drugs, have been found to be active in a number of solid and hematologic malignancies. Three topoisomerase I inhibitors have been investigated in the treatment of metastatic or recurrent squamous carcinoma of the head and neck: 9-aminocamptothecin (9-AC), topotecan (Hycamtin), and irinotecan (CPT-11, Camptosar). Neither 9-AC nor topotecan has demonstrated clinically significant activity in the treatment of metastatic or recurrent squamous carcinoma of the head and neck. In contrast, irinotecan has demonstrated a modest overall response rate of 21.2% (95% confidence interval [CI] = 9% 38.9%), with a median survival of 214 days and a 1-year survival rate of 30.2%. The response and toxicity appear to be dose dependent. Further investigation of irinotecan in combination with other active agents and radiotherapy is warranted. PMID- 11497233 TI - Irinotecan in relapsed or refractory non-Hodgkin's lymphoma. AB - Irinotecan (CPT-11, Camptosar) is a topoisomerase I inhibitor with a broad spectrum of antitumor clinical activity. Various schedules and doses have been studied, and major complications were delayed diarrhea and myelosuppression. We explored the activity of irinotecan in patients with relapsed or refractory non Hodgkin's lymphoma, using a 3-week schedule of administration. Eligible patients had histologically proven relapse, had received no more than two previous regimens, were > or = 15 years and < or = 75 years old, had normal renal function, neutrophil count > 1,500/microL, platelet count > 100,000/microL, and no human immunodeficiency virus infection or central nervous system involvement. Patients were treated with irinotecan 300 mg/m2 i.v. every 21 days with intensive loperamide management of diarrhea. Responders received up to six treatment cycles. Of 25 patients registered so far, 22 are evaluable for response. The median age was 67 years (range: 25 to 74 years) and 11 were male. The median number of previous regimens was 2 (range: 1 to 4 regimens), and 16 patients had disease that was refractory to their last regimen. Serum lactate dehydrogenase level was high in 75%, and beta2-microglobulin was > 3.0 mg/L in 26% of patients. Responses were seen in 8 of 22 (36%) patients with non-Hodgkin's lymphoma. Response rates were 40% for indolent, 0% for mantle cell, 45% for relapsed aggressive, and 33% for refractory aggressive lymphomas. Grade 3/4 toxicities included myelosuppression, neutropenic fever, and delayed diarrhea. Irinotecan appears active and relatively well tolerated in patients with relapsed aggressive non-Hodgkin's lymphoma. Accrual to this study is continuing for better determination of the response rate in all histologic subtypes of non-Hodgkin's lymphoma. PMID- 11497234 TI - Role of topoisomerase I inhibitors in small-cell lung cancer. AB - Combination chemotherapy is the cornerstone of treatment that confers a meaningful survival benefit for patients with small-cell lung cancer. However, because there have been no major therapeutic advances for small-cell lung cancer during the last decade, more effective new treatments are necessary to improve the outcome of therapy. Irinotecan (CPT-11, Camptosar), a topoisomerase I inhibitor, is one of the new active agents that provide hope for more effective therapies. In single-agent phase II studies, irinotecan yielded response rates between 16% and 47% in patients with previously treated small-cell lung cancer. A phase II study of irinotecan in combination with cisplatin (Platinol) resulted in a response rate of 86% and a median survival of 13.0 months in patients with extensive-disease small-cell lung cancer. A phase III trial that was conducted by the Japan Clinical Oncology Group (JCOG) clearly demonstrated a survival advantage for the combination of irinotecan and cisplatin vs the standard regimen of etoposide (VP-16, VePesid) and cisplatin. Based on these results, the irinotecan and cisplatin combination is a new standard regimen in the treatment of extensive-disease small-cell lung cancer. PMID- 11497235 TI - Werner syndrome: genetic and molecular basis of a premature aging disorder. AB - Werner syndrome (WS) is a rare autosomal recessive disorder characterized by genomic instability and by the premature onset of a number of age-related diseases, including cancers. The gene responsible for WS encodes a protein that has an exonuclease domain and a domain similar to DNA helicases of the RecQ-like subfamily. Accumulating evidence indicates that the WS gene product is involved in resolving aberrant DNA structures that may arise during the process of DNA replication and transcription. Such processes generate regions of single-stranded DNA that may inadvertently provide a substrate for the initiation of recombination. Various mechanisms have evolved to ensure that recombination does not occur promiscuously during these events, and results are consistent with a model in which the WS protein is part of one such mechanism. PMID- 11497236 TI - Mass spectrometry for protein and peptide characterisation. AB - Mass spectrometry has become an important analytical tool in biological and biochemical research. Its speed, accuracy and sensitivity are unmatched by conventional analytical techniques. Identification of proteins and characterisation of their primary structure is a rapidly growing field in the post-genomic era, where matrix-assisted laser desorption/ionisation time-of flight peptide mass fingerprinting combined with electrospray tandem mass spectrometry can efficiently solve many questions. Many recently determined genomic sequences have not been characterised at the protein level. Analysis of the amino acid sequence and characterisation of post-translational modifications are therefore important steps towards correlation of protein structure with function. This review concerns methods, instrumentation and applications of mass spectrometry in protein and peptide analysis. PMID- 11497237 TI - Structural studies of discoidal lipoprotein A-I. AB - Apolipoprotein A-I (apoA-I) is a major exchangeable apolipoprotein of high density lipoproteins (HDLs), and plays an important role in reverse cholesterol transport. This process involves transport of cholesterol from peripheral tissues to the liver for processing, thereby eliminating excess cholesterol from the body. The function of apoA-I and its interaction with other components of HDL, including lecithin-cholesterol acyltransferase, seems to be closely linked to its structural plasticity. ApoA-I is likely to undergo changes in its structure and orientation between the various HDL subclasses and, therefore, knowledge of the precise structure of apoA-I is essential for understanding its role in the antiatherogenic properties of HDL. This review focuses on the role of apoA-I in reverse cholesterol transport and the work done by various groups to determine the structure of apoA-I in discoidal HDL particles. PMID- 11497238 TI - RecQ helicases and topoisomerases: components of a conserved complex for the regulation of genetic recombination. AB - Maintenance of genomic stability relies on the efficient and accurate execution of DNA repair pathways, and is essential for cell viability and the prevention of cancer. Mutation of genes encoding RecQ helicases or topoisomerases gives rise to genomic instability through excessive recombination. Here, we review the recent biochemical and genetic evidence to indicate that these two classes of protein act in concert in a conserved pathway to maintain genomic stability by preventing inappropriate recombination. PMID- 11497239 TI - Multifunctional alpha-enolase: its role in diseases. AB - Enolase, a key glycolytic enzyme, belongs to a novel class of surface proteins which do not possess classical machinery for surface transport, yet through an unknown mechanism are transported on the cell surface. Enolase is a multifunctional protein, and its ability to serve as a plasminogen receptor on the surface of a variety of hematopoetic, epithelial and endothelial cells suggests that it may play an important role in the intravascular and pericellular fibrinolytic system. Its role in systemic and invasive autoimmune disorders was recognized only very recently. In addition to this property, its ability to function as a heat-shock protein and to bind cytoskeletal and chromatin structures indicate that enolase may play a crucial role in transcription and a variety of pathophysiological processes. PMID- 11497240 TI - AMPA receptors: potential implications in development and disease. AB - Alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) receptors are one type of ionotropic glutamate receptor involved in rapid excitatory synaptic transmission. AMPA receptors have been increasingly implicated in long-term potentiation, and recent evidence suggests that they may play a role in disorders affecting the nervous system. The finding that early in postnatal development AMPA receptors are not expressed has lately been the focus of much attention. Resolving the factors involved in AMPA receptor expression suggests that their induction is a developmentally regulated process with the possibility that alterations in receptor expression may be correlated with pathology in neurological disorders. This paper provides an overview of factors involved in AMPA receptor induction as well as of their role in plasticity and neuronal pathologies. PMID- 11497241 TI - From MDR to MXR: new understanding of multidrug resistance systems, their properties and clinical significance. AB - The ATP binding cassette (ABC) superfamily of membrane transporters is one of the largest protein classes known, and counts numerous proteins involved in the trafficking of biological molecules across cell membranes. The first known human ABC transporter was P-glycoprotein (P-gp), which confers multidrug resistance (MDR) to anticancer drugs. In recent years, we have obtained an increased understanding of the mechanism of action of P-gp as its ATPase activity, substrate specificity and pharmacokinetic interactions have been investigated. This review focuses on the functional characterization of P-gp, as well as other ABC transporters involved in MDR: the family of multidrug-resistance-associated proteins (MRP1-7), and the recently discovered ABC half-transporter MXR (also known as BCRP, ABCP and ABCG2). We describe recent progress in the analysis of protein structure-function relationships, and consider the conceptual problem of defining and identifying substrates and inhibitors of MDR. An in-depth discussion follows of how coupling of nucleotide hydrolysis to substrate transport takes place, and we propose a scheme for the mechanism of P-gp function. Finally, the clinical correlations, both for reversal of MDR in cancer and for drug delivery, are discussed. PMID- 11497242 TI - Regulatory RNAs. AB - In addition to mRNA, rRNA and tRNA, which play central roles within cells, there are a number of regulatory, non-coding RNAs (ncRNAs). Of varying lengths, ncRNAs have no long open reading frame. While not encoding proteins, they may act as riboregulators, and their main function is posttranscriptional regulation of gene expression. Many ncRNAs have been identified and characterized both in prokaryotes and eukaryotes, and are involved in the specific recognition of cellular nucleic acid targets through complementary base pairing, controlling cell growth and differentiation. Some are associated with the abnormalities in imprinted inheritance that occur in several well-known developmental and neurobehavioral disorders. Other ncRNAs accomplish regulation by modulating the activity of proteins. Several rRNAs are able to sustain enzymatic reactions implicated in the translation process including synthesis of peptide bonds within the ribosome. The different roles played by widely distributed RNAs acting in diverse ways, suggest the flexibility and versality of these molecules in regulatory processes. This review summarizes the available biochemical and structural data on known regulatory RNAs. PMID- 11497243 TI - The role of mammalian antimicrobial peptides and proteins in awakening of innate host defenses and adaptive immunity. AB - Since we live in a dirty environment, we have developed many host defenses to contend with microorganisms. The epithelial lining of our skin, gastrointestinal tract and bronchial tree produces a number of antibacterial peptides, and our phagocytic neutrophils rapidly ingest and enzymatically degrade invading organisms, as well as produce peptides and enzymes with antimicrobial activities. Some of these antimicrobial moieties also appear to alert host cells involved in both innate host defense and adaptive immune responses. The epithelial cells are a source of constitutively produced beta defensin (HBD1) and proinflammatory cytokine-inducible beta defensins (HBD2 and -3) and cathelicidin (LL37). The neutrophils-derived antimicrobial peptides are released on demand from their cytoplasmic granules. They include the enzymes cathepsin G and chymase, azurocidin, a defensins and cathelicidin. In contrast, C5a and C3b are produced by activation of the serum complement cascade. The antimicrobial moieties direct the migration and activate target cells by interacting with selected G-protein coupled seven-transmembrane receptors (GPCRs) on cell surfaces. The beta defensins interact with the CCR6 chemokine GPCRs, whereas cathelicidins interact with the low-affinity FPRL-1 receptors. The neutrophil-derived cathepsin G acts on the high-affinity FMLP receptor (GPCR) known as FPR, while the receptors for chymase and azurocidin have not been identified as yet. The serum-derived C5a uses a GPCR known as C5aR to mediate its chemotactic and cell-activating effects. Consequently, all these ligand-receptor interactions in addition to mediating chemotaxis also activate receptor-expressing cells to produce other mediators of inflammation. PMID- 11497244 TI - The effect of hydrogen peroxide on the cyclin D expression in fibroblasts. AB - Activation of mitogen-activated protein (MAP) kinase is essential for cyclin D1 expression and provides a link between mitogenic signalling and cell cycle progression. Hydrogen peroxide (H2O2) activates MAP kinase; however, it is not known whether this leads to cyclin D expression. Sustained expression of cyclin D1 and D2 was observed when Her14 fibroblasts were incubated with 3 mM or higher H2O2 concentrations. Similar results were obtained when cells were incubated in the presence of serum (FCS). However, the sustained expression of cyclin D1 and D2 upon H2O2 treatment was not due to the MAP kinase pathway, because MAP kinase kinase inhibitors did not inhibit cyclin D expression. Furthermore, cyclin D1 and D2 levels remained constant even after addition of a protein synthesis inhibitor, indicating that the effect of H2O2 was not due to induction of protein synthesis. These results indicate that H2O2 reversibly inhibits the ubiquitin-proteasome dependent degradation of cyclin D1 and D2, probably by transiently inhibiting ubiquitination and/or the proteasome. PMID- 11497245 TI - PM-3, a benzo-gamma-pyran derivative isolated from propolis, inhibits growth of MCF-7 human breast cancer cells. AB - Propolis has numerous biologic activities including antibiotic, antifungal, antiviral and anti-inflammatory properties. Several components isolated from propolis have been shown to have anticancer activity. This study demonstrates that the compound PM-3 (3-[2-dimethyl-8-(3-methyl-2-butenyl)benzopyran]-6 propenoic acid) isolated from Brazilian propolis markedly inhibits the growth of MCF-7 human breast cancer cells. This effect was associated with inhibition of cell cycle progression and induction of apoptosis. Treatment of MCF-7 cells with PM-3 arrested cells in the G1 phase and resulted in a decrease in the protein levels of cyclin D1 and cyclin E. PM-3 also inhibited the expression of cyclin D1 at the transcriptional level when examined in cyclin D1 promoter luciferase assays. Induction of apoptosis by PM-3 occurred within 48 hours after treatment of MCF-7 cells. The MCF-7 treated cells also displayed a decrease in the level of the estrogen receptor (ER) protein and inhibition of estrogen response element (ERE) promoter activity. Therefore, PM-3 merits further investigation with respect to breast cancer chemoprevention or therapy. PMID- 11497246 TI - Targeting of LAK activity to CEA-expressing tumor cells with an anti-CEA scFv/IL 2 fusion protein. AB - BACKGROUND: Fusion of tumor-specific monoclonal antibody (MAb) and cytokines has proved to be an efficient way to target cytokines to tumor cells and hence focuses the killing activity of effector cells to the target cells. We previously produced a high affinity MAb, F11-39, against carcinoembryonic antigen (CEA), which is often overexpressed on the surface of various tumor cells. MATERIALS AND METHODS: To target the cytotoxicity of effector cells to CEA-expressing tumor cells, we employed recombinant DNA techniques to fuse recombinant human interleukin-2 (rhIL-2) to a single chain variable fragment (scFv) antibody derived from F11-39. The resulting fusion protein, designated F39scFv/IL-2, was expressed in the Sp2/0-Ag14 mouse hybridoma cells, purified by CEA-affinity chromatography and characterized for the CEA-binding specificity and the IL-2 biological activity. RESULTS: F39scFv/IL-2 protein effectively targeted rhIL-2 onto the surface of CEA-expressing tumor cells and consequently introduced a specific cytotoxicity of lymphokine-activated killer cells to the tumor cells. CONCLUSIONS: This approach may be used for in vivo administration to localize IL 2 to tumor tissues, maximizing the immune response to CEA-expressing tumors while keeping systemic side effects to a minimum. PMID- 11497247 TI - High anticancer efficacy of L-proline-m-bis (2-chloroethyl) amino-L-phenylalanyl L-norvaline ethyl ester hydrochloride (MF13) in vivo. AB - The anticancer efficacy of the new anticancer tripeptide, L-proline-m-bis (2 chloroethyl) amino-L-phenylalanyl-L-norvaline ethyl ester hydrochloride (MF13), was investigated in mice. MF13 showed a therapeutic effect in liquid tumors and induced complete remission even in late stage malignancies. MF13 also inhibited human colon cancer growth in nude mice by more than 85% (volume, p<0.001). It acted in a dose-dependent manner and induced a complete regression of tumor in 20% of the mice when the initial dose was high (15 mg/kg, i.p.). Human melanoma exhibited a response to MF13 similar to colon cancer. Activity of MF13 in murine hepatoma in vivo was stronger than its precursor m-sarcolysin (p<0.001). Tumor cells in peritoneal cavities of the MF13 treated (s.c.) mice underwent an irreversible apoptosis. Side effects of MF13 were the transient depression of hemopoiesis and loss of body weight, which vanished within 9-10 days. LD50 of MF13 of a single i.p. injection was 27 mg/kg (94 mg/m2), 11 times higher than the therapeutic dose of a single injection. PMID- 11497248 TI - Effect of high retinoid doses on RAR-beta mRNA expression in female B6D2F1 mice. AB - The expression of retinoid receptors is altered during the development of several types of cancer. In the present study, we determined the influence of high dietary concentrations of 4-hydroxyphenylretinamide (4-HPR) and 13-cis-retinoic acid (13-cis-RA) on RAR-beta mRNA expression in female mice. Expression of liver and lung RAR-beta RNA increased with increasing levels of dietary retinoid (both 4-HPR and 13-cis RA). Bladder RAR-beta mRNA levels, however, were significantly decreased in mice fed 13-cis RA or 4-HPR. These results suggest that feeding high levels of retinoids to mice results in tissue-specific elfects on expression of RAR-beta mRNA. PMID- 11497249 TI - uPA and uPAR contribute to NK cell invasion through the extracellular matrix. AB - BACKGROUND: The urokinase plasminogen activator (uPA) system has been implicated in cellular invasiveness of tumor cells and immune cells. Herein we provide evidence for the production by natural killer (NK) cells of both uPA and its receptor (uPAR). MATERIALS AND METHODS: Western blot analysis, RTPCR, casein/plasminogen zymography, and fluorescence microscopy were employed to detect uPA and uPAR on NK cells. NK cell invasiveness was examined using Matrigel invasion assays. RESULTS: NK cell uPA appeared at its characteristic molecular weights, is enzymatically active in casein/plasminogen zymography, and is recognized by monoclonal antibodies. uPAR was detected by RTPCR and fluorescence microscopy. Matrigel invasion assays demonstrated an active role of uPA in NK cell invasion. CONCLUSION: The uPA system contributes to extracellular matrix (ECM) degradation by NK cells, which may be essential for NK cell accumulation into metastases, and may be prerequisite for their killing of tumor cells following NK cell adoptive transfer. PMID- 11497250 TI - Possible regulation of 5-fluorouracil-induced neuro- and oral toxicities by two biochemical modulators consisting of S-1, a new oral formulation of 5 fluorouracil. AB - S-1 is a new oral formulation of 5-fluorouracil (5-FU) containing 1 M tegafur and 0.4 M 5-chloro-2,4-dihydroxypyridine (CDHP) and 1 M potassium oxonate (Oxo). It has been reported to have a high antitumor activity and low gastrointestinal toxicity in rats bearing murine and human tumors. We further studied the possible inhibition of the toxicities caused by the products of 5-FU metabolism with the use of CDHP, a new inhibitor of 5-FU degradation and Oxo, an inhibitor of 5-FU phosphorylation. In a model of pentylenetetrazole-induced convulsions in mice, intravenous injection of fluoroacetate (3 mg/kg), 2-fluoro-b-alanine (30 mg/kg) and 5-FU (over 300 mg/kg) significantly augmented the occurrence of convulsion. However coadministration of an equivalent dose of CDHP with 5-FU almost completely suppressed the 5-FU-augmented convulsions, suggesting that inhibition of 5-FU catabolism by CDHP may lead to a decreased risk of development of 5-FU neurotoxicity. Another advantage of the use of S-1 was protection through Oxo against the development of 5-FU-induced mucositis, which occurs frequently in cancer patients. When 6 mg/kg of S-1 was administered orally to beagle dogs for 5 days, the incidence of stomatitis decreased markedly compared to that in dogs receiving the same dose of S-1 not containing Oxo, in which severe stomatitis was frequently observed. One of the possible mechanisms of the decreased incidence of mucositis associated with oral S-1 administration is the decreased formation of 5 fluoronucleotides from 5-FU in the mucosal tissues of the oral cavity. These results suggest that oral S-1 could be employed for the treatment of cancer patients with marked reduction in the incidence of toxicities including encephalopathy, stomatitis and diarrhea. PMID- 11497251 TI - Effects of 10-hydroxycamptothecin, delivered from locally injectable poly(lactide co-glycolide) microspheres, in a murine human oral squamous cell carcinoma regression model. AB - This study investigated whether local delivery of 10-hydroxycamptothecin provides effective inductive chemotherapy as assessed by significant tumor reduction. Established tumorigenic human oral squamous cell carcinoma cells were used for these experiments. The experimental groups were comprised of: control (blank (no drug) poly(lactide-co-glycolide) (PLGA) microspheres), intraperitoneal 10 hydroxycamptothecin delivery + blank microspheres, local bolus 10 hydroxycamptothecin + blank microspheres, and PLGA controlled-release microspheres. The 10-hydroxycamptothecin dose administered was 12 mg/kg (bolus intraperitoneal, local) or controlled-release over 10 days. Regardless of delivery route, 10-hydroxycamptothecin significantly reduces tumor volume. However, PLGA microspheres provide significantly higher intratumor-drug concentrations (approximately 10 and 100 fold higher) relative to local bolus and intraperitoneal routes, respectively. Also, only the PLGA microspheres significantly reduced tumor weights. Camptothecin clinical applications are limited by drug inactivation at physiological pH and the need for sustained infusions. However, due to their acidic, camptothecin-stabilizing microclimate, PLGA microspheres could provide a novel delivery system for camptothecin-based induction chemotherapy. PMID- 11497252 TI - Bone morphogenetic protein-2 and type IV collagen expression in psammoma body forming ovarian cancer. AB - Psammoma bodies (PBs), characterized as calco-spherules with concentric laminations, are common in serous tumors of the ovary. However, there is no agreements as to how the PBs are formed. Bone morphogenetic protein-2 (BMP-2) has recently been proposed to be involved in the calcification of tumor cells and recent electron microscopic studies demonstrated the presence of type IV collagen in PBs. Based on this evidence, we postulated a possibe role for BMP-2 and type IV collagen in the formation of PBs in ovarian cancer. We examined the expression of BMP-2 and typle IV collagen by immunohistochemistry and reverse transcription PCR (RT-PCR) in PBs-forming (NK-211) and -non-forming (SHIN-3, KF-1, A2780, KK 92, KOC-2S, SKOV-3, OMC-3, MN-1, EC, and KEN-3) ovarian cancer cell lines in vitro and in surgical specimens of serous adenocarcinoma (SA) with/without PBs and mucinous adenocarcinoma (MA) of the ovary. Cellular growth of cell lines was also evaluated by their doubling time in vitro. Transcripts for BMP-2 mRNA were detected by RT-PCR in all cell lines. By immunohistochemistry, BMP-2 protein expression was positive in 45% (5 out of 11) of cell lines. 36.4% (4 out of 11) were positive for type IV collagen. PBs-forming NK-211 was intensively positive for both BMP-2 and type IV collagen. In addition, NK-211 demonstrated extremely slow growth with a doubling time of 450 hours. In surgical specimens, BMP-2 vs. type IV collagen positivities in tumor cells were 100% (20 out of 20) vs. 40% (8 out of 20) in SA with PBs, 61.1% (11 out of 18) vs. 0% (0 out of 18) in SA without PBs and 75% (9 out of 12) vs. 0% (0 out of 12) in MA. In PBs themselves, 100% (20 out of 20) positivity for BMP-2 and 80% (16 out of 20) for type IV collagen was shown. These results raise the possibility that BMP-2 and type IV collagen-producing slow growing tumor cells form PBs in ovarian cancer. PMID- 11497253 TI - Identification of metastasis-associated genes by transcriptional profiling of a metastasizing versus a non-metastasizing human melanoma cell line. AB - In order to identify genes associated with the metastatic phenotype we have compared the expression pattern of 6800 genes in a metastatic (NMCL-1) versus a non-metastatic (530) human melanoma cell line making use of DNA microarrays. The differentially expressed genes identified are involved in control of transcription, regulation of the cell-cycle, proteolysis, cell adhesion, immune response and signaling. A remarkable feature of the system under investigation is the consistent down-regulation of MHC-related and cell adhesion mediating genes in the metastatic cell line. PMID- 11497254 TI - Nanoerythrosomes, a new derivative of erythrocyte ghost: IV. Fate of reinjected nanoerythrosomes. AB - Recently, we have developed a promising new drug carrier named nanoerythrosome (nEryt). This transporter are small vesicles made with the red blood cell membrane. Anticancer drugs like daunorubicin, linked to these nEryt, have a higher antineoplastic activity than the free drug. In this paper, we first analyzed the biodistribution of 125I-nEryt purified by dialysis following intravenous (i.v.) or intraperitoneal (i.p.) injections in CD1 mice. After i.v. administration, nEryt, are rapidly removed from blood circulation (< 30 min). Mainly the liver and spleen take up the vesicles. I.p. injections of nEryt purified by dialysis, showed a marked activity in the inguinal lymph nodes 2 hours post-injection. nEryt purified by centrifugation have a different biodistribution. They accumulate also in the lungs. We demonstrated that accumulation in the lungs is due to particle aggregation during the preparation procedure. Comparative analysis of size distribution of each nEryt preparation revealed that nEtyt purified by centrifugation has a mean diameter of 1.5 microm which is 10 times higher than its dialyzed counterpart. Light microscopic autoradiographs of dialyzed nEryt, reinjected i.v., showed accumulation of nEryt in the sinusoidal lumen as well as in the parenchymal cells of the liver. Autoradiographs of the spleen revealed that nEryt are distributed specifically near the marginal zone and that some of them have escaped the meshes of the red pulp cords. PMID- 11497255 TI - Indomethacin reduces lung adenoma number in A/J mice. AB - The effects of indomethacin on A/J mice were investigated. The non-steroidal antiinflammatory drug (NSAID) indomethacin reduced significantly the number of lung adenomas 3, 4 or 8 months after urethane injection by 28, 30 and 29% respectively. The density of apoptotic cell bodies increased 2.9-fold in the lung adenomas of A/J mice treated with indomethacin. By immunocytochemistry, COX-2 immunoreactivity was present in the cytosol of lung adenomas, and in epithelial cells lining the bronchioli and bronchus as well as type 2 alveolar cells. COX-1 immunostaining was similar to that of COX-2 in the lungs of urethane-injected mice treated with or without indomethacin. By RT-PCR, COX-1 and COX-2 PCR products were present in mouse lung adenomas, alveoli and bronchioli. These results suggest that indomethacin may inhibit COX-1 and COX-2 in the A/J mouse lung resulting in reduced adenoma formation. PMID- 11497256 TI - Localization of metallothionein in urothelial carcinoma of the human urinary bladder: an immunohistochemical study including correlation with HLA-DR antigen, p53, and proliferation indices. AB - Metallothionein (MT) is a low-molecular-weight cysteine-rich protein, which has the ability to bind and sequestrate heavy metal ions. Synthesis of MT is induced in a variety of tissues by these metal ions, as well as by endogenous factors such as glucocorticoids, interferon, interleukin-1 and vitamin D. Several lines of evidence show that MT may play a role in carcinogenesis. In this study MT expression was detected immunohistochemically, using a monoclonal antibody (E9) against a conserved epitope of I and II isoforms, in a series of 63 cases of urothelial carcinomas of the urinary bladder. Correlation between MT expression and HLA-DR antigen expression, p53, proliferation indices (PCNA and MIBI) as well as the various clinicopathological parameters, such as age, sex, squamous metaplasia, tumor grade, stage and recurrence were studied. In a semiquantitative analysis MT expression (> 10% of neoplastic cells) was observed in 12.7%, focal MT positivity in 11.1% and almost completely lack of MT expression in 76.2% of tumors. The incidence of MT expression was significantly higher (p=0.0002) in cases with high pathological tumor grades. MT values were significantly correlated with tumor stage (p=0.0009). A statistically significant positive correlation between MT expression and the HLA-DR antigen expression (p=0.001) was also detected. The data suggested that MT expression was correlated with a more aggressive behavior in urothelial bladder cancer. PMID- 11497257 TI - Discovery of novel inducers of cellular differentiation using HL-60 promyelocytic cells. AB - Non-physiological inducers of terminal differentiation have been used as novel therapies for the prevention and therapy of cancer. We have used cultured HL-60 promyelocytic cells to monitor differentiation, proliferation and cell death events as induced by a large set of extracts derived from plants. Screening of more than 1400 extracts led to the discovery of 34 with potent activity (ED50 <8 mg/ml). Bioassay-guided fractionation led to the isolation of zapotin and 2',5,6 trimethoxyflavone as active principles from Casimiroa edulis, dibenzyltrisulfide and 2-[(phenylmethyl)dithio]ethanol as active principles from Petiveria alliacea, and desmethylrocaglamide from Aglaia ponapensis. Zapotin demonstrated the most favorable biological profile in that induction of differentiation correlated with proliferation arrest, and a lack of cytotoxicity. We conclude that the HL-60 cell model is a useful system for the discovery of novel pharmacophores with potential to suppress the process of carcinogenesis, and that flavonoids may be especially useful in this capacity. PMID- 11497258 TI - Identification of metastasis-associated genes by transcriptional profiling of a pair of metastatic versus non-metastatic human mammary carcinoma cell lines. AB - Cell lines 4A4 and 2C5 are the respective metastatic and non-metastatic variants of the human mammary carcinoma cell line MDA-MB-435 in the nude mouse system. We compared the transcriptional profile of approximately 5000 full-length genies using the Affymetrix HuGene FL Array technology. We have shown that the metastatic phenotype is mediated by different functional categories of genes, e.g. genes involved in immune response, genes responsible for tumor antigens, genes involved in migration and invasion, genes involved in mediating signal transduction, genes responsible for transcription factors, genes involved in phospholipid signaling, genes involved in modulation of extracellular matrix and cytoskeleton, genes with a cell-type specific mode of expression and genes which do not fit into the subclasses as defined above. Our results suggest an important role of Autocrine Motility Factor (AMF) as a mediator of metastasis in this system. PMID- 11497259 TI - Inhibition of growth of human breast carcinoma cells by an antisense oligonucleotide targeted to the transferrin receptor gene. AB - Transferrin receptor expression is controlled by the amount of iron required by the cell to maintain its metabolism and therefore tumor cells in a highly proliferative state have a high density of transferrin receptors. In this study, phosphorothioated antisense TfR oligonucleotides (TfR-ODna) targeted to the sequences of TfR mRNA including the AUG initiation codon and the control sense chain (TfR-ODns) were synthesized. The rate of cellular DNA synthesis was determined by [3H]-thymidine incorporation. Administering TfR-ODna to three morphologically distinct breast cancer cell lines (MCF-7, T47D, and MDA-MB-231) and a normal breast cell line (MCF-12A) caused specific inhibition of tumor cell growth. The IC50 (50% inhibition of DNA synthesis) of the TfR-ODna for the MCF-7, T47D and MDA-MB-231 cells were 0.5, 0.5, and 1.0 microM, respectively, whereas the MCF-12A normal breast cells were about 30 times (IC50 of 30 microM) less sensitive to TfR-ODna than the breast cancer cells. The cytotoxicity of the antisense TfR-ODna was 10 to 60 times greater than that of the sense chain (TfR ODns). TfR mRNA and protein synthesis were demonstrated by RT-PCR and immunohistochemical staining, respectively. Approximately 50% inhibition of the expression of TfR mRNA was observed when breast cancer cells were treated with 1 microM antisense TfR ODNa for 72 hrs but 1 microM antisense only caused 14% inhibition in normal breast cells. The decreased cytotoxicity and inhibition of TfR gene expression when the tumor cells were treated with the same concentration (1 microM) of TfR-ODns demonstrated the specificity of the TfR-ODna for blocking the target TfR gene. The combined cytotoxicities to human breast tumor MCF-7 cells of the antisense TfR-ODna and the iron chelator deferoxamine (DFO) or the ribonucleotide reductase inhibitor hydroxyurea were observed in this study. IC50s (50% inhibition of DNA synthesis) for DFO and hydroxyurea individually were 0.3 microM and 250 microM, respectively. The CalcuSyn program was used to determine the combined effects among the agents and synergism (Combined Indexes (CI) < 1) were found with the following two combinations: TfR-ODna (0.007 microM to 0.15 microM) with DFO (0.15 microM to 5 microM) and TfR-ODna (0.007 microM to 0.15 microM) with hydroxyurea (50 microM to 800 microM). However, inhibition by TfR ODns was not synergistic with either DFO or hydroxyurea. The synergistic effects on inhibition of DNA synthesis between TfR-ODna and DFO or hydroxyurea suggest that inhibition of breast cancer cell growth by TfR-ODna is produced by depletion of iron pools that are required for DNA synthesis in tumor cells. The fact that TfR-ODna specifically decreases cell viability and proliferation, and reduces TfR mRNA and protein expression in human breast carcinoma cells without affecting normal breast cells, suggests that the antisense oligonucleotide to the transferrin receptor may be a novel therapeutic approach in breast cancer. PMID- 11497260 TI - Irofulven (6-hydroxymethylacylfulvene, MGI 114) induces caspase 8 and 9-mediated apoptosis in human pancreatic adenocarcinoma cells. AB - BACKGROUND: Irofulven (MGI 114) is a novel, clinically active sesquiterpene whose mechanism of action is not fully understood. We sought to identify apoptotic effectors induced by this agent in human pancreatic cancer cells. MATERIALS AND METHODS: MTT assay was used to assess IC50-Apoptosis was quantitated by flow cytometry and DAPI staining. Caspase activation was identified by western blot analysis. RESULTS: Irofulven was cytotoxic against all pancreatic cancer cell lines tested (IC50 1-18 microM), and induced 10-fold (4%+/- 2, vs. 41% +/- 5) induction of apoptosis. Irofulven-treated cells also demonstrated PARP3 cleavage and DAPI staining. Apoptosis was reduced to baseline levels by Z-VAD-FMK, a broad spectrum caspase inhibitor. Western blot analysis revealed that caspases-3, -7, 8, and -9 were activated by irofulven. Time course evaluation demonstrated that caspases-8 and -9 were the initial species activated. CONCLUSION: Our data demonstrate that the cytotoxicity of irofulven in human pancreatic carcinoma cell lines is mediated by caspase-induced apoptosis. PMID- 11497261 TI - Influence of extracellular pH on the cytotoxicity and DNA damage of a series of indolequinone compounds. AB - BACKGROUND: The existence of an acidic extracellular pH (pHe) within solid tumours is regarded as a potential target for drug development. The indolequinone EO9 has a complex mechanism of action which includes enhanced potency under acidic pHe conditions in vitro. In order to identify compounds which have a simpler mechanism of action where activation under acidic pHe is the predominant mechanism of toxicity, this study has determined the cytotoxic properties of a series of analogues of EO9 under both physiological and acidic pHe conditions. MATERIALS AND METHODS: H460 human NSCLC cells were exposed to EO compounds under acidic (pH 6.04) and physiological (pH 7.24) pHe conditions for one hour and chemosensitivity assessed 4 days later using the MTT assay. For compounds of interest, DNA damage (both single strand breaks and cross links) in H460 cells was determined using the comet assay. RESULTS: All the compounds tested were more potent under acidic pHe conditions although a broad range of enhancement ratios (defined as the IC50 at pHe 7.24/IC50 at pHe 6.04) were obtained ranging from 3.25 to 116.53. The activity of EO72 was significantly enhanced under acidic conditions and activity was associated with a pH dependent increase in DNA cross linking in H460 cells. As EO72 is a poor substrate for purified human DT diaphorase, pHe conditions appear to be a major factor determining cell kill. CONCLUSIONS: This study has identified several compounds whose cytotoxic properties in vitro are pHe dependent with EO72 emerging as the lead compound on the basis of the magnitude of the pH dependent chemosensitivity and the fact that it is a poor substrate for DT-diaphorase. Further studies are required to determine whether or not EO72 has suitable pharmacokinetic properties to allow it to reach regions of low pHe within solid tumours. PMID- 11497262 TI - Growth inhibition of mouse skin tumor by serine protease inhibitor ONO-3403. AB - The new serine protease inhibitor, ONO-3403 is an analog of FOY-305 (Foypan). The IC50s values of ONO-3403 toward serine proteases, such as trypsin, plasmin, kallikrein and thrombin are much lower than that of FOY-305. To investigate the growth-suppressing effect of ONO-3403 on 3-methylcholanthrene-induced autochthonous solid tumors in mice, ONO-3403 was intraperitoneally administered to mice at a dose of 4 mg/kg twice a day for 5 weeks. All seven mice receiving the drug had a solitary tumor and showed potent growth suppression (p<0.001) without any apparent side effects such as hair loss and body weight loss. The results suggest that ONO-3403 may be useful for the treatment of squamous cell carcinoma. PMID- 11497263 TI - A new antitumour treatment combining radiation and electric pulses. AB - AIM: To investigate the antitumour effect of radiation in combination with electropermeabilization on subcutaneous rat glioma tumours. MATERIALS AND METHODS: Sub-optimal radiation treatment was administered separately or in combination with electric pulses of high voltage to subcutaneous rat brain tumours. The treatment was repeated on four consecutive days and evaluated by TGD and microscopical examination. The tumours were stained for Factor VlII/von Willebrand Factor to investigate the effects on the tumour vasculature. RESULTS: Radiation and electric pulses applied concomitantly resulted in a cure rate of 67% (tumour free >80 days after treatment). Radiation-treated animals showed progressive disease. Histological and immunohistochemical examination of electric impulse-treated tumours showed instant and severe deteriorating effects on tumour vasculature. CONCLUSION: A distinct antitumour effect of the combined treatment of electric pulses and radiation treatment was observed. We believe that the tumouricidal effect arises from destruction of the tumour vasculature but also from DNA related damage from reactive oxygen formed by the electric pulses and the radiation treatment. PMID- 11497264 TI - Electrically mediated drug delivery for treatment of an adenocarcinoma transplanted into rat liver. AB - BACKGROUND: In this study, electrochemotherapy (ECT), i.e. tumour treatment based on local augmentation of intracellular drug delivery from short, intense electric pulses, was evaluated in rats with an adenocarcinoma implanted into the liver. Tumour response and concentrations of macrophages and T-lymphocytes (CD4 and CD8) in and around the tumour were measured. MATERIALS AND METHODS: Rats were treated with permeabilizing electric pulses, bleomycin, or both, eight days after implantation of the tumour, while one group received sham treatment. RESULTS: Treatment with electric pulses and bleomycin resulted in a significantly reduced lesion volume and 92% cure rate (12 out of 13, p<0.0002 compared to the other treatment groups). The highest concentration of CD8 lymphocytes was found in tumours treated with electric pulses and bleomycin. Macrophages were found mainly in tumours treated with electric pulses, with or without bleomycin. CONCLUSION: Electrochemotherapy using millisecond exponential pulses and bleomycin is efficient in a rat liver tumour model and appears to stimulate the host's immune system. PMID- 11497265 TI - 9-Nitrocamptothecin is an effective drug for the treatment of human lung tumors: comparison of in vitro and in vivo studies. AB - 9-Nitrocamptothecin (9NC) results in complete regression of small-cell lung carcinoma (SCLC) and non-SCLC (NSCLC) growing as xenografts in immunodeficient mice. In this study, we have monitored histological changes in the tumors during 9NC-induced regression, and perturbations in the cell cycle of cells derived from these tumors using flow cytometry. In vivo, 9NC treatment induces dramatic changes in the tumor cells, which die by apoptosis and are ultimately eliminated from the normal tissue. In vitro, 9NC treatment resulted in apoptosis and cytostasis of the NSCLC and SCLC cells, respectively. Further, 9NC induced cytostasis in control, normal human lung fibroblasts. Therefore, the studies in vivo have indicated that 9NC acquires a remarkable antitumor activity against both the SCLC and NSCLC types tested, and that results of studies in vitro may not reflect the results observed in vivo. PMID- 11497266 TI - Hypermethylation does not account for the frequent loss of the retinoic acid receptor beta2 in breast carcinoma. AB - Hypermethylation of the retinoic acid receptor (RAR) beta2 has been detected in breast cancer cell lines and is known to repress the level of RAR beta2 transcription. RAR beta2 mRNA loss has often been detected in breast cancer tumors, whether promoter region methylation of the RAR beta2 gene accounts for its loss is still unknown. We examined the methylation status of RAR beta2 in breast tumors; 21 out of 50 (42%) breast tumors showed RAR beta2 hypermethylation. RT-PCR analysis showed a complete loss of RAR beta2 mRNA expression in 15 out of 43 (35%) breast tumors. No correlation between the hypermethylation and RAR beta2 loss was found, suggesting that hypermethylation is not fully responsible for the loss of expression of the RAR beta2 gene during breast tumorigenesis. PMID- 11497267 TI - Efficacy of single-bolus vs. frequent low-dose treatment with nitrosourea in experimental gliomas. AB - BACKGROUND: Malignant glioma remains a fatal disease. Continuous or frequent low dose (FLD) chemotherapy with nitrosoureas reportedly causes fewer side-effects than single-bolus therapy without decreasing the antitumour effects. MATERIALS AND METHODS: To study the effect of FLD treatment with nimustine (ACNU) in rats with glioma, we intracerebrally inoculated with C6 glioma cells. We began the ACNU treatment 5 or 8 days later (total dose, 25 or 40 mg/kg) i.p. as either one bolus or smaller doses spread over 5 days week. RESULTS: At a total dose of 25 mg/kg beginning at day 8, survival duration did not differ between untreated controls and the FLD group, while the bolus significantly prolonged survival; the FLD group showed some improvement beyond control survival at 40 mg/kg (each p <0.001). Beginning treatment after 5 rather than 8 days prolonged survival somewhat further. CONCLUSION: FLD treatment with ACNU is less effective against experimental glioma in rats than bolus treatment. PMID- 11497268 TI - Embryonic apoptosis-inducing proteins exhibited anticancer activity in vitro and in vivo. AB - A 54 Kd apoptosis-inducing protein with novel amino acid sequence has been purified from the conditioned medium of the embryonic cell line, C3H 10T1/2 cells. An apoptosis-inducing protein identified to be fetal fetuin and a 60 Kd apoptosis-inducing protein have also been found in fetal serum and fresh embryo extract, respectively. Interestingly, a common characteristic of these embryonic apoptosis-inducing proteins is that they selectively induced apoptosis in cancer without affecting normal cells. For example, the 54 Kd protein selectively induced apoptosis in 10 out of 12 cancer cell lines without affecting 12 normal cell lines we tested. Fetal fetuin, on the other hand, selectively induced apoptosis in 5 cancer cell lines without affecting the 3 normal cell lines we tested. In vivo, tumor animal model study showed that fetal fetuin enhanced survival in leukemia-bearing mice and strongly inhibited the formation of prostate cancer in a PC-3 prostate cancer model in mice. A working hypothesis has been proposed to aid in the study of the mechanism by which the embryonic apoptosis-inducing proteins selectively induced apoptosis in cancer without affecting normal cells. This hypothesis states that due to the retro differentiational characteristic of malignancy, cancer cells may re-express the signal transduction machinery for development-related apoptosis, which is otherwise to be normally expressed by embryonic, but not by adult cells. The embryonic apoptosis-inducing proteins may therefore induced apoptosis in cancer but not in normal cells and may be developed as an anticancer agent. This new concept may constitute a new approach for cancer therapy, which we tentatively designated as "Retro-differentiational Apoptosis Cancer Therapy", (R-ACT). PMID- 11497269 TI - Cytotoxicity and mode of action of 1-(1-cyclohexenyl) and 1-unsubstituted 3,5 pyrazolidinediones in human Molt4 T cell leukemia. AB - The 3,5-pyrazolidinediones proved to be potent cytotoxic agents against the growth of a number of murine and human tumor cell lines, e.g. human THP-I monocytic leukemia, Hut-78 lymphoma, MCF-7 breast effusion, A549 lung carcinoma, U87MG glioma, Hela uterine and A431 epidermoid skin cancer. In human Tmolt4 cell leukemia, the agents substantially suppressed DNA and RNA syntheses after 60 min at 100 microM. The de novo purine biosynthetic pathway appeared to be the major target of the agents with the inhibition of both PRPP-amido transferase and IMP dehydrogenase (IMPDH) activities. Suppression of IMPDH activity was due to the inhibition of both the Type I and II isoforms through an uncompetitive mechanism; however, the Type II isoform was preferentially inhibited at lower concentrations of compounds tested (>50-150 microM). Therefore IMPDH Type II activity, which predominates in cancer cells, was selectively inhibited over the Type I isoform (208-312 microM). The activities of other enzymes examined were inhibited which added to the overall suppression of DNA synthesis, i.e., ribonucleotide reductase, dihydrofolate reductase and nucleoside kinases. The agents caused Tmolt4 DNA strand scission but the DNA molecule itself did not appear to be a target of the compounds since there was no induced cross-linking of the DNA, intercalation between base pairs or alkylation of the DNA bases. PMID- 11497270 TI - In vivo inhibition by antioxidants of adriamycin-induced apoptosis in murine peritoneal macrophages. AB - Adriamycin (ADM) is an oncostatic of the anthracycline family with confirmed experimental and clinical efficiency. This antitumoral drug has been reported to stimulate macrophage activity and is able to induce apoptosis (AP) in some tumour cells. The objective of the present work was to investigate if in vivo administration of ADM to mice induces AP in their peritoneal macrophages (PM). AP was expressed by the apoptotic index (AI) of peritoneal macrophages observed under fluorescence microscope after ethidium bromide and acridine orange staining and confirmed by detection of the ladder pattern on DNA electrophoresis, indicates DNA fragmentation in 80-120 bp characteristic of apoptotic state. 24 hours after i.p. ADM administration, AP was observed in PM. The effect was best visible after the injection of 5 mg/kg ADM. (Al: 76.3+/-8.9 vs untreated control group AI: 2.8+/-1.1). In the ADM treated group a DNA ladder electrophoretic pattern was observed while DNA from normal PM was genomic. Since ADM toxicity has been attributed to reactive oxygen species generation, we investigated its possible participation in AP induction by pretreating mice with antioxidants: (+) alpha-tocopherol acid succinate (30 IU/mouse per os) for 3 days before ADM administration with E. coli lipopolysacharide (0.15 microg/mouse i.p.) 24 hours before ADM administration or with superoxide dismutase (10,000 IU/mouse i.p.) 1 hour before ADM administration. AI was significantly decreased, with values close to those of the untreated control group (AI: 15+/-5.7, 9.6+/-8.0 and 32.9+/-6.9, respectively). Antioxidants given before ADM treatment significantly increased the live cell index (p < or = 0.001) in PM the groups while inactivated antioxidants no longer protect PM against the ADM AP induction. DNA analysis confirmed the effect: in the untreated control and in the antioxidant protected groups DNA was genomic while in either ADM or inactivated-antioxidants + ADM treated groups, DNA presented the ladder pattern. AP can thus be induced in PM by ADM and inhibited by antioxidants. These observations may have clinical applications. PMID- 11497271 TI - Immunocytochemical localization of basic fibroblast growth factor in squamous cell carcinomas of the head and neck. AB - Angiogenesis is crucial for tumour growth and formation of metastasis. Basic fibroblast growth factor (bFGF) is known to have potent angiogenic activity and has been identified in a wide variety of malignancies including head and neck squamous cell carcinomas (HNSCC). Clarification of its localization in HNSCC is important for the understanding of angiogenesis. Cryosiat sections of 27 HNSCC were immunostained for bFGF using a standard streptavidin-biotin complex procedure. Western blot analysis revealed three immunoreactive bFGF isoforms of 18, 22 and 24 kDa. Immunohistochemical and -cytochemical localization of bFGF was studied at light and electron microscopic levels. bFGF was mainly localized within the focal tumoral areas rather than in the tumour stroma. On a subcellular level, the ultrastructural investigation showed electron-dense bFGF localization in the cytosol of the carcinoma cells, but bFGF labelling within the nuclei of HNSCC cancer cells was one prominent finding of this study. In conclusion, the presence of bFGF isoforms in most of the cancer cells supports the theory of a direct paracrine mechanism of bFGF from cancer cells on tumour angiogenesis. The nuclear localization of bFGF in the HNSCC cells supports its activity as a transcriptional factor. PMID- 11497272 TI - Modification of apoptosis-inducing activity of gallic acid by saliva. AB - Gallic acid, a structural unit of tannin, induced apoptotic cell death, characterized by nuclear condensation and caspase activation in human oral tumor cell lines (HSC-2, HSG). Agarose gel electrophoresis demonstrated that gallic acid produced large DNA fragments in these cells, as well as in T-cell leukemia (MOLT-4) and erythroleukemia (K-562) cells, whereas it induced internucleosomal DNA fragmentation in human myelogenous leukemic cell lines (HL-60, ML-1, U-937, THP-1). This indicates that induction of internucleosomal DNA fragmentation or production of large DNA fragments depends on both target cells and inducers. Addition of total saliva dose-dependently reduced the cytotoxicity induction by gallic acid. These data suggested that the biological action of gallic acid might be modified by physiological fluids. PMID- 11497273 TI - Serum stimulation and cell density regulate the proliferation of AsPC-1 cells through control of cyclin E and p27KIP1 expression. AB - BACKGROUND: Cellular proliferation in normal cells is tightly regulated by environmental conditions. Growth factors stimulate proliferation while cell confluence inhibits it. Human pancreatic cancer AsPC-1 cells were believed to escape from these restrictions because they possessed several mutations which promote cell proliferation. In this study, we focused on the relationships between growth conditions and the proliferation of AsPC-1 cells. MATERIALS AND METHODS: AsPC-1 cells were cultured under several growth conditions and the proliferation of cells was studied by incorporation of 3H-thymidine. The alterations of cell-cycle-related genes were studied by immunoblotting. RESULTS: By four consecutive days in culture, the nucleotide incorporation of AsPC-1 cells was markedly suppressed and the suppression was overcome by medium change or reduction of cell density. The induction of cyclin D1 by serum stimulation was observed, concomitant with the transient activation of extracellular signal regulated kinases (ERKs). The most prominent changes of cell-cycle-regulating genes following consecutive culture or serum reduction were the down-regulation of cyclin E and the induction of p27KIP1. The down-regulation of cyclin E was more sensitive to cell density, while the induction of p27KIP1 was regulated by both increased cell density and reduction of serum. The down-regulation of p27KIP1 was caused by protein degradation. CONCLUSIONS: The proliferation of AsPC 1 cells was still controlled by cell density and serum stimulation; nevertheless, the cells possessed several oncogenic mutations. These results may provide a rationale for modifying the growth environment for treatment of pancreatic cancers. PMID- 11497274 TI - Tumour cell uptake G2-M accumulation and cytotoxicity of NAMI-A on TS/A adenocarcinoma cells. AB - The ruthenium(III) complex imidazolium trans-imidazoledimethylsulfoxide tetrachlororuthenate (NAMI-A) was tested on TS/A adenocarcinoma cells to evaluate the relationship between cell uptake, cell cycle arrest and cytotoxicity. The in vitro challenge of TS/A cells with 10(-4) M NAMI-A for 15 minutes to 4 hours showed a partial reduction of cell growth only after 4 hour exposure. In the same experimental conditions NAMI-A caused the increase of cells in G2-M cell cycle phase directly proportional on the length of treatment, and the ruthenium uptake by tumour cells, measured by flameless atomic absorption spectroscopy, that increases up to 2 hours of treatment and then reaches a plateau. The arrest of cell cycle in the pre-mitotic G2-M phase was transient and completely reversed by 48 hours after treatment. This study showed that the effect of NAMI-A on the cell cycle of TS/A cells is not strictly related to NAMI-A uptake as is the effect on tumour cell proliferation. PMID- 11497275 TI - Alterations in DNA repair and telomere maintenance mechanism affect response to porphyrins in yeast. AB - BACKGROUND: DNA quadruplex-interactive porphyrin TMPyP4, but not its isomer TMPyP2, inhibits telomerase activity and causes chromosome fusion in vivo, suggesting interference with telomere maintenance. MATERIALS AND METHODS: We examined effects of these porphyrins and hydroxyurea on growth rates of yeast Saccharomyces cerevisiae wild type and strains with defects in telomere maintenance and/or DNA repair pathways (mec1, tel1, rad9), telomere binding protein (cdc13), and anaphase control (pds1). RESULTS: Hydroxyurea (20 mM) decreased proliferation rates only in mec1 mutant and deletion strains. TMPyP4 (200 microM) decreased growth in all strains, especially in rad9delta and mec1delta. The growth inhibition by TMPyP4 showed low growth inhibition in strains defective in cdc13 and pds1. TMPyP2 sterically prevented from forming a planar species did not significantly inhibit growth of any strain. Overexpression of telomere binding protein Rap1 hypersensitized the mec1delta and tel1delta to TMPyP4. CONCLUSIONS: Telomere maintenance represents a viable target for anticancer agents. PMID- 11497276 TI - Antiproliferative effect of isopentenylated coumarins on several cancer cell lines. AB - 33 coumarins, mainly the simple isopentenylated coumarins and derived pyrano- and furanocoumarins, were examined for their antiproliferative activity towards several cancer and normal human cell lines. The pyrano- and furanocoumarins showed strong activity against the cancer cell lines, whereas they had weak antiproliferative activity against the normal human cell lines. The decreasing rank order of potency was osthenone (10), clausarin (25), clausenidin (26), dentatin (24), nordentatin (23), imperatorin (29), seselin (27), xanthyletin (21), suberosin (17), phebalosin (8) and osthol (12). The structure-activity relationship established from the results revealed that the 1,1-dimethylallyl and isopentenyl groups have an important role for antiproliferative activity. PMID- 11497277 TI - Effect of a synthetic analog of prostaglandin E1 on the intestinal mucosa of methotrexate-treated rats. AB - Administration of methotrexate to rats sometimes induces small intestinal damage. A synthetic analog of prostaglandin E1, OP-1206 [17S,20-dimethyl-trans-delta2 prostaglandin E1] may possibly provide therapeutic benefits to help recovery from such small intestinal damage. The purpose of this study was to evaluate the effect of OP-1206 on methotrexate-induced small intestinal damage in rats. Methotrexate (15 mg/kg body weight) was orally administered to rats once daily for 5 days. OP-1206 (0.5 microg/kg body weight) was orally administered to rats twice a day for 5 days and on the 6th day the small intestine of the rats were examined histologically and biochemically. The methotrexate treatment of rats caused a severe histological change in the small intestinal mucosa, whereas the treatment combined of OP-1206 with methotrexate showed similar histological features of the small intestinal mucosa as that of the control rats. On the other hand, an acute intestinal inflammation was evaluated by determining myeloperoxidase activity. The myeloperoxidase activity in the small intestinal mucosa of the methotrexate-treated rats increased remarkably, whereas that of the methotrexate and OP-1206-treated rats was significantly lower than that of the methotrexate-treated rats. Thus, it was shown histologically and biochemically that OP-1206 was effective in protecting the small intestine from methotrexate induced damage. PMID- 11497278 TI - Mechanism of growth-inhibitory effect of cisplatin on human pancreatic cancer cells and status of p53 gene. AB - Pancreatic cancer is a devastating malignant tumor in humans and the development of new modalities of treatment is needed. We studied the mechanism of the growth inhibitory effect of cisplatin (CDDP) on human pancreatic cancer cells in connection with the status of the p53 gene and expression of the bcl-2 family. COLO-357 cells with wild-type p53 gene and T3M4, Panc-1 and AsPC-1 cells with mutant-p53 gene were used. Growth of these cells was inhibited by CDDP in a dose dependent manner in both serum-deprived and serum-supplemented conditions. CDDP induced apoptosis of COLO-357 and T3M4 cells in the serum-supplemented condition, whereas necrosis of these cells was induced by CDDP at high concentrations in the serum-deprived condition. Although expression of bax mRNA and its protein product were enhanced, while bcl-2 protein was decreased by CDDP in COLO-357 cells, expression of mRNA of the bcl-2 family and protein product were not influenced by CDDP in T3M4 cells. Increased expression of bax and reduced expression of bcl-2 are involved in the growth-inhibitory effect of CDDP on pancreatic cancer cells with wild-type p53 gene. PMID- 11497279 TI - Potentiation of the antiproliferative effect in vitro of doxorubicin, cisplatin and genistein by new analogues of vitamin D. AB - Numerous vitamin D3 analogues have been synthesised in recent years in order to obtain compounds with a favourable biological and therapeutic (antipsoriatic and/or antitumour) activity. Our results showed that pre-treatment for 72 hours of HL-60 human promyelocytic leukaemia cells with calcitriol or its new analogues significantly potentiated their sensitivity to the antiproliferative effect in vitro of cisplatin, doxorubicin or genistein. Moreover, for all cytotoxic agents tested a synergistic antiproliferative effect was observed. This effect was expressed as a significant decrease of the ID50 (inhibitory dose 50%) values for each cytotoxic agent applied after pretreatment with calcitriol or its analogues of HL-60 cells in comparison with the effect of cytotoxic agent applied alone. The observed in vitro potentiated antiproliferative effect of cytotoxic drugs used in combination with vitamin D or its analogues may raise the question as to whether such an effect could be expected in the in vivo situation. PMID- 11497280 TI - Clinical significance of the Rb expression in adenocarcinoma of the lung. AB - BACKGROUND: Altered retinoblastoma gene expression has been found in various types of cancers including non-small cell lung cancer (NSCLC) and some reports have shown it also to be associated with a poor clinical prognosis in NSCLC. MATERIALS AND METHODS: We evaluated the expression of Rb protein by immunohistochemistry in 90 patients with lung adenocarcinoma who underwent surgical resection and determined its prognostic significance. Also, the expression of p27 by immunohistochemistry and the mutations of the p53 gene by PCR-SSCP were evaluated. RESULTS: Fifty-one out of 90 tumors (56.7%) had reduced expression of the Rb gene. There were no statistical differences between the Rb expression level and clinicopathological status. There was no correlation between the Rb expression level and the p53 mutations or the p27 expression level. The 5 year survival rate in the normal group was 55.1%, which was poorer than that of 73.0% in the reduced group, but showed no statistical difference (p=0.0981). In patients with a reduced Rb-expression, the survival rate of the p53 mutation group tended to show a poorer prognosis than that of the p53 normal group (p=0.0880). In addition, the survival rate of the negative p27 expression group tended to show a poorer prognosis than that of the positive p27 expression group (p=0.0537). CONCLUSIONS: The Rb expression status was not significant as an individual factor for evaluating the prognosis in adenocarcinoma of the lung. Analyses of the accumulation of genetic alteration are necessary to identify the subset of patients with a poor prognosis. PMID- 11497281 TI - FADD gene therapy using the human telomerase catalytic subunit (hTERT) gene promoter to restrict induction of apoptosis to tumors in vitro and in vivo. AB - Gene transfer vectors will dramatically increase the safety and effectiveness of cancer gene therapy, if they could restrict expression of the therapeutic products to the target tumors. To realize such a tumor-targeting system, telomerase is one of the most promising candidates. It is because telomerase activity is detected in the vast majority of tumors, but not in most normal cells. Activation of telomerase is tightly regulated at the transcriptional level of the telomerase catalytic subunit (hTERT). Therefore, the use of the hTERT promoter-driven vector system could restrict the expression of therapeutic products to telomerase-positive tumors. In this study, we constructed the expression vector of FADD gene with death domain afforded by the hTERT promoter (hTERT/FADD) and investigated its effect on tumors in vitro and in vivo. Transient transfection with the hTERT/FADD construct induced apoptosis in telomerase-positive tumor cells of wide range. In contrast, normal fibroblast cells without telomerase did not undergo apoptosis following the hTERT/FADD transfer. Furthermore, the growth of subcutaneous tumors in nude mice was significantly suppressed by the intratumoral injection of the hTERT/FADD construct (every day for one week) compared to the control (P<0.0005). The findings described here indicate the high potentiality of a novel telomerase specific gene therapy of tumors with telomerase. PMID- 11497282 TI - Inhibition of vascular endothelial cells by 1,4-phenylenebis (methylene)selenocyanate--a novel chemopreventive organoselenium compound. AB - Organoselenium compound 1,4-phenylenebis(methylene)selenocyanate (p-XSC) was investigated for its effects on endothelial cell proliferation in vitro and angiogenesis in vivo. The organoselenium compound, p-XSC, has been shown to prevent carcinogen-induced tumorigenesis in murine model systems with low toxicity. Since tumor growth and metastasis are dependent on angiogenesis, we investigated the effects of the organoselenium compound on this process. Human umbilical vein endothelial cells treated with p-XSC showed concentration dependent inhibition of protein synthesis and cell viability in vitro with a TCID50 value of 6 microM. Subsequently, we studied the effects of p-XSC on experimental angiogenesis. Addition of p-XSC to three-dimensional cultures inhibited endothelial cell tube formation. Furthermore, p-XSC treatment inhibited growth factor induced angiogenesis in chick chorioallantoic membrane assays and i.p. administration of p-XSC inhibited neovascularization induced by tumor cells implanted subcutaneously into athymic mice. These studies suggest that vascular endothelium is an additional target for the chemopreventive organoselenium compound p-XSC. PMID- 11497283 TI - Characterization of ten newly-derived human head and neck squamous carcinoma cell lines with special reference to c-erbB proto-oncogene expression. AB - Ten human head and neck squamous carcinoma (HNSCC) cell lines were established in order to study the role of c-erbB signaling pathways in HNSCC progression. Five cell lines were derived from primary tumors at four different sites, and five were from lymph node metastases in the neck. Two pairs of lines were derived from the primary tumor and metastatic lymph node in the same patient. Basic characteristics including morphology, doubling time, phenotypes, cytogenetic profiles and tumorigenicity in nude mice were described. We examined the expression of c-erbB receptors and ligands in early passage new HNSCC lines and compared with five long-term established lines, normal keratinocytes and fibroblasts. Amplification of c-erbB-1 (EGFR) gene was observed in only one cell line whereas no amplification of other c-erbB genes was found. Overexpression of EGFR, c-erbB-2, c-erbB-3 and c-erbB-4 mRNAs was observed in 10, 14, 10 and 8 out of 15 head and neck cell lines respectively. Overexpression of c-erbB-3 and c erbB-4 was more frequently observed in newly derived HNSCC lines than in long established cell lines. The majority of tumor cells also expressed multiple c erbB ligands. One selected cell line, SIHN-006, was shown to exhibit tyrosine phosphorylation via all four receptors. These new cell lines could provide a useful experimental model to study the co-operative signaling of type I tyrosine kinase receptors in HNSCC progression. PMID- 11497284 TI - Anticancer activity of an aqueous mistletoe extract (AME) in syngeneic murine tumor models. AB - Mistletoe extracts have been used for decades for non-specific stimulation of the immune system in cancer therapy. Mistletoe lectins (ML) have been identified as the active principle with cytotoxic and immunomodulatory potencies. In the present in vivo experiments, the anticancer effects of an aqueous mistletoe extract (AME) were investigated in different subcutaneously growing syngeneic murine tumors such as Renca renal cell carcinoma, C8 colon 38 carcinoma, F9 testicular carcinoma, B16 melanoma and Lewis lung carcinoma. The animals used were immunocompetent mice of different strains (C57BL/6, BALB/c), depending on the type of tumor tested. After tumor transplantation, the mice were treated with AME at dose levels corresponding to 0, 0.3, 3, 30 or 300 ng ML/kg/d by the i.p. or s.c. route for a maximum of 4 consecutive weeks. The tumor volume was determined by serial caliper measurements and expressed relative to controls. Significant tumor growth inhibition was observed with the Renca , C8 colon 38 and F9 testicular carcinomas at 30 and 300 ng ML/kg/d. These findings were confirmed in independent repeat experiments. No inhibitory effects were seen with the Lewis lung carcinoma and B16 melanoma under the conditions described above. In conclusion, AME showed in vivo anticancer activity in different transplantable syngeneic murine tumor models following repeated parenteral treatment. In view of the low dose levels used, the effects are most likely due to the immunostimulatory rather than to the cytotoxic potencies of AME. PMID- 11497285 TI - Growth suppression activity of the PTEN tumor suppressor gene in human endometrial cancer cells. AB - The AKT proteins are constitutively activated in several types of human cancers, which may play a role in carcinogenesis. In this study, we examined the activation of AKT in a panel of human endometrial cancer cell lines and tumor samples in this study. Two endometrial cancer cell lines, Ishikawa (ISK) and RL 95 and several tumor samples showed elevated levels of phosphorylated AKT PTEN, which is mutated in 45% of endometrial cancers, is a negative regulator of AKT. We examined the growth suppression activity of PTEN in ISK and KLE endometrial cancer cells. Expression of PTEN significantly suppressed the growth of both cell clines. In primary rat embryo fibroblasts, PTEN also inhibited malignant transformation mediated by ras and c-myc oncogenes. These two oncogenes are commonly mutated or amplified in endometrial cancer. These results suggest that PTEN may be a potent therapeutic agent for endometrial cancer. PMID- 11497286 TI - Aurintricarboxylic acid decreases proliferative potential of SKOV3 and MCF7 human carcinoma cells. AB - The effect of aurintricarboxylic acid (ATA) on cell growth and proliferative capacity was studied in human ovarian SKOV3 and breast MCF7 carcinoma cells. ATA moderately inhibited cell growth measured by a Neutral red assay after a 24-hour incubation of the cells in the presence of ATA. The ATA-treated cells displayed a markedly decreased capacity to proliferate, as was evident from a colony formation assay. The initial and delayed anti-proliferative effects of ATA were dose-dependent. Together, the results indicated that ATA offers the potential of being recognized as an anti-tumor drug, at least in certain types of cancers. PMID- 11497287 TI - Antineoplastic activity of novel N-1-sulfonypyrimidine derivatives. AB - We evaluated the potential activity of novel N-1-sulfonyl derivatives of pyrimidine bases uracil and cytosine on pancreatic carcinoma cells (MIAPaCa2), colon carcinomas cells (HT-29, CaCo2), cervical carcinoma cells (HeLa) and poorly differentiated cells from lymph node metastasis of colon carcinoma (SW-620). The cytotoxicity of N-1-sulfonylpyrimidine derivatives was analyzed with the MTT cell survival assay and their antiproliferative activity was measured via radioactive precursors incorporation assay. The N-1-sulfonylpyrimidine derivatives affected the growth of all examined cell lines at concentrations of 10(-8)-10(-5) M, by 25 70%. Growth inhibition depended on the tumor cell line type and the concentration of investigated compounds. The compounds 2, 4, 7, 8 and 9 inhibited DNA, RNA and protein synthesis in CaCo2, MIAPaCa2 and HeLa cells. The exposure of tumor cells in vitro to compounds 2, 4, 7, 8, 9, 10 and 11, at the 10(-6) M concentration, caused both morphological (condensation of chromatin, cell shrinkage), as well as biochemical changes (ladder pattern of DNA fragmentation and exposure of phosphatidylserine on outer lipid bilayer plasma membrane) characteristic of apoptosis. After 24 hours of the N-1-sulfonylpyrimidine derivative application, the p53 oncoprotein expression could not be detected by immunocytochemical analysis. On the basis of present results it can be concluded that novel N-1 sulfonylpyrimidine derivatives are promising antitumor agents with a strong antiproliferative activity and an ability to induce apoptosis in treated tumor cells. PMID- 11497288 TI - Time sequential expression of markers of apoptosis induced by 1-beta-D arabinofuranosylcytosine in human T-lymphoblastic leukemia CCRF-CEM cells. AB - A wide variety of chemotherapeutic agents including 1-beta-D arabinofuranosylcytosine can induce cell death by apoptosis. However, an appropriate method for early detection of apoptosis that can be clinically applicable to peripheral blood samples freshly obtained from leukemic patients under chemotherapy has not yet been established. We investigated the chronology of ara-C induced apoptosis in CCRF-CEM cells by monitoring the expression of a number of apoptotic markers in a time- and concentration-dependent manner with the aim of estimating the earliest reliable marker of apoptosis to apply to clinical drug sensitivity assays. We employed the following techniques: 1) Pulsed field gel electrophoresis to detect high molecular weight DNA fragmentation; 2) FACS analysis for evaluation of APO 2.7 expression and phosphatidylserine externalization; 3) May-Grunwald-Giemsa staining for studying gross morphology; and 4) TUNEL assay to detect multitudes of terminal 3'-OH groups of oligonucleosomal DNA fragments. At 10 microM ara-C, high molecular weight DNA fragmentation was observed after 4 hours incubation being the earliest marker to manifest. APO 2.7 expression and phosphatidylserine externalization were detected almost simultaneously at 6 hours, with marked similarity in their kinetics. Emergence of apoptotic bodies then followed after 12 hours incubation and, finally, oligonucleosomal DNA fragments were demonstrated by positive TUNEL assay at 48 hours. The results suggest the time-sequential expression of each individual marker and introduce high molecular weight DNA fragmentation assay as the most suitable candidate for early detection of sensitivity of malignant cells to apoptosis-inducing anticancer agents, including ara-C. PMID- 11497289 TI - The carcinostatic and proapoptotic potential of 4-hydroxynonenal in HeLa cells is associated with its conjugation to cellular proteins. AB - BACKGROUND: Previous studies have shown that the lipid peroxidation product 4 hydroxynonenal (HNE) acts as a cell growth modulator if used at low, physiological concentrations being strongly cytotoxic at higher concentrations for a number of cells. These effects of HNE also appeared to be mutually dependent on the effects of serum growth factors. The aim of this investigation was to study the concentration-dependent response of human cervical carcinoma (HeLa) cells in vitro with respect to the intracellular uptake of exogenous HNE, the cellular energy metabolism, DNA synthesis, overall gene expression and susceptibility to apoptosis. MATERIALS AND METHODS: MTT assay was applied as an index of energy metabolism and the replicative activity was quantitated by the 3H thymidine incorporation assay. The occurence and intracellular distribution was studied with monoclonal antibodies directed against HNE-protein conjugates. Binding of HNE to serum proteins was determined with the same antibodies by Western blotting. Differential gene expression was studied by differential display RT-PCR while a novel photometric assay, denoted Titer-TACS, was used for in situ detection and quantitation of apoptosis in monolayer cell cultures. RESULTS: A physiological concentration of HNE (1 microM) had hardly any effect on the parameters of the replicative activity and the energy metabolism. No morphological changes were observed and the number of HNE-positive cells was not significantly different when compared to the untreated control cells, while most of the aldehyde appeared to be bound to serum proteins (albumin fraction). A ten fold higher concentration (10 microM) was found to be cytostatic. Spindle-shaped cells with a picnotic nucleus were observed occasionally, as well as membrane blebs, which were HNE-positive. The number of HNE-positive cells was significantly increased compared both to the control cells and cells treated with 1 microM HNE, but in the presence of serum the effects of 10 microM HNE were negated due to its binding to the serum proteins. Finally, 100 microM HNE was cytotoxic for the HeLa cells. Most of the cells were picnotic, together with a few spindle-shaped or oval cells. The staining for HNE was diffuse and strong (90% of the cells were HNE-positive) while even binding of the aldehyde to serum proteins did not prevent its cytotoxic effects. This concentration of HNE caused acute stress response of the cells resulting in the decreased expression of several as yet unidentified genes. The altered pattern of gene expression was followed by programmed cell death, i.e. an increased number of apoptotic cells after treatment with low (1 and 10 microM) concentrations of HNE. A rebound effect was observed, i.e. a decrease of apoptotic cells after 24 hours followed by an overshooting increase after 48 hours. CONCLUSIONS: For HeLa carcinoma cells there appears to be a concentration range of HNE where it does not cause necrosis but preferentially apoptosis. At this concentration range HNE is cytochemically detectable within the cells as a protein conjugate. It is proposed that a possible differential sensitivity of cancer cells and their normal counterparts to the cytostatic activity of HNE should be explored. PMID- 11497290 TI - Growth regulation effects of gonadotropin induced steroidogenic response in human ovarian cancer. AB - BACKGROUND: Despite a substantial body of epidemiological evidence, there is only a limited indication that gonadotropins and steroids have growth regulating functions in ovarian cancer. To elucidate the role of gonadotropins in regulating steroid metabolism in human ovarian cancer, we analyzed the modulation of estradiol secretion by FSH and hCG and the gonadotropic regulation of hCG secretion in vitro. Furthermore, we analyzed estradiol and hCG levels in serum and cyst fluids of patients with ovarian cancer. MATERIALS AND METHODS: OVCAR3 cells were incubated with estradiol (1, 5, 10 nM), FSH (100 microg/L) and hCG (10, 25 microg/L). Growth stimulation was evaluated by MTT assay. Estradiol was measured in the supernatant after incubation with hCG and FSH, while hCG was measured after FSH incubation. FSH, estradiol and hCG levels were measured in serum and cyst fluids of patients with ovarian cancer. RESULTS: OVCAR3 cells responded to hCG and FSH by increased estradiol secretion (p<0.001), while estradiol led to a dose-dependent stimulation of cell growth (p<0.05). 100 microg/L FSH led to a 75% decrease of hCG secretion (p<0.001). CONCLUSION: Gonadotropins stimulate estradiol secretion in ovarian cancer cells and modulate steroid dependent growth stimulation. FSH modulates hCG related growth stimulation in ovarian cancer. These results were supported by in vivo measurements in ovarian cancer patients. PMID- 11497291 TI - Localization of human BRCA1 and BRCA2 in non-inherited colorectal carcinomas and matched normal mucosas. AB - We characterized the expression of BRCA1 and BRCA2 in 38 sporadic colorectal carcinomas and matched normal mucosas with 9 anti-BRCA1 antibodies and 4 anti BRCA2 antibodies, raised against several different epitopes, using immunohistochemical technique. We demonstrated an increased BRCA1 and BRCA2 staining in the apical cell pole of epithelial malignant cells and we also revealed a significant increase in BRCA1 and BRCA2 nuclear foci in tumor colorectal specimens in comparison with corresponding normal tissues. These increases in BRCA1 and BRCA2 expression may be explained by the fact that colorectal tissue is subject to very active proliferation and differentiation. PMID- 11497292 TI - Matrix metalloproteinases in neoplasm-induced extracellular matrix remodeling in breast carcinomas. AB - Structural changes in the extracellular matrix (ECM) are necessary for cell migration during normal and pathologic tissue remodeling and neoplastic cell invasion. The matrix metalloproteinases (MMPs) and their inhibitors have been identified to be critical modulators of ECM composition and are thus, crucial in neoplastic cell progression, invasion and metastasis. Expression of MMP-2, -3, 9, -10, and -13 was investigated in human breast carcinomas (BCs) employing an indirect, biotin-streptavidin based, alkaline phosphatase conjugated immunocytochemical technique. Evaluation of the results was based on (a) the percent of neoplastically transformed cells/surrounding stroma that reacted positively and (b) a measure of staining intensity [graded from A (highest) to D (negative)]. The two forms of stromelysin, MMP-3 and -10, share 82% sequence homology, but exhibit differences in cellular synthesis and inducibility by cytokines and growth factors in vitro. Strong overall expression of MMP-3 and -10 was found in BCs, especially in the ECM adjacent to blood vessels. Positive immunoreactivity could be seen for these two MMPs in the ECM surrounding over 90% of the neoplastically transformed cells (++++), and the staining intensity was also the strongest possible (A). High intensity immunoreactivity (A,B) but focal was detected employing a MoAB targeted against the MMP-9 enzyme. No presence of MMP-2 or -13 could be established in the BC cases observed by us. Based on these results we propose that MMP-3 and -10 are implicated in the pathogenesis of BC, while MMP-9 is possibly involved in neo-angiogenic events also closely associated with growth and expansion of the neoplastically transformed cell mass, as well as metastasis of individual, extremely aggressive, expressing dedifferentiated cellular immunophenotype (IP) cell clones selected during the microevolution of the BC. PMID- 11497293 TI - Effect of pneumoperitoneum on circulating tumor DNA. AB - BACKGROUND: Release and circulation of tumor DNA could be favored by surgery. No data is available for the effect of laparoscopy on this phenomenon. MATERIAL AND METHODS: The aim of this study was to assess the impact of CO2 laparoscopy on circulating tumor DNA. Two xenografts of ovarian cancer were obtained by intraperitoneal inoculation (IP) of IGR-OV1 or NIH:OVCAR-3 cells in nude rats. CO2 laparoscopy (L), gasless laparoscopy (GL), midline laparotomy (ML) or general anesthesia as a control (C) were randomly carried out when the tumor graft was present in the peritoneal cavity. A sterile blood sample was taken in each case as soon as the experiment was completed. DNA was subsequently extracted and amplified (PCR, primers HLA GH 26 and HLA GH 27 specific for human DNA). In each model, we compared the influence of each surgical approach on circulating tumor DNA. Statistics were performed with the Wilcoxon test and Fisher exact test. 1: RESULTS: Eighteen rats were included in each group. Our protocol could detect an amount of tumor DNA equivalent to 10 cells/ml of blood. This technique was specific. Circulating tumor DNA was frequently observed in the IGR-OV1 model (45 to 50%), without significant difference between groups (p=0.6). In the NIH: OVCAR 3 model, the detection rate ranged from 22% (control group) to 64% (gasless group); but the overall comparison between the four groups was not significant (p = 0.2). CONCLUSION: In this experimental trial, CO2 laparoscopy had no deleterious effect on circulating tumor DNA. Biologic characteristics of tumors could also play a role. PMID- 11497294 TI - Polyaspartamides as water-soluble drug carriers. Part 1: Antineoplastic activity of ferrocene-containing polyaspartamide conjugates. AB - BACKGROUND: Numerous detrimental side effects and/or lack of water-solubility of anticancer drugs often prove dose-limiting in chemotherapy. Water-soluble polymeric drug carriers may overcome/minimise many of these limitations. MATERIALS AND METHODS: Aspartic acid polymers to which ferrocene-containing antineoplastic agents are covalently bound, were tested for cytotoxicity against murine EMT-6 cancer cells. Cell survival was measured by means of the colorometric 3-(4,5-dimethylthiazol-2-yl)-diphenyltertrazolium bromide assay. RESULTS: The 90% lethal dosage of pure 3-ferrocenylbutanoic acid is 452 microg/mL LD90 for the polymeric derivative, expressed in terms of 3-ferrocenylbutanoic acid content, is only 65 microg/ml. A polymer structural effect in drug activity was evident: longer side chains linking drugs to polymer backbones enhanced drug activity. Drug activity is also enhanced if drug modifications (to enable drug anchoring) resulted in a lower ferrocenyl reduction potential. CONCLUSIONS: The effectivity of antineoplastic drugs may be enhanced by covalently anchoring them on suitable biodegradable water-soluble polymeric drug carriers. PMID- 11497295 TI - Cellular accumulation determines the activity of three novel tricyclic platinum agents. AB - BACKGROUND: Cisplatin (CDDP) is a very useful chemotherapeutic agent, but cellular resistance limits its efficacy in malignant glioma. In order to analyze and overcome this resistance, we synthesized three novel platinum compounds; aminoplatin, methylplatin and oxiplatin, using tricyclic DNA intercalating molecules as models. MATERIALS AND METHODS: The novel compounds differ only in one chemical group, which is positioned opposite to the DNA binding site. DNA binding, cellular penetration, and cytotoxicity were compared in three human glioma cell lines (T98G, U87-MG, and U25 IN) and a human fibroblast cell strain (HS 68). 2 RESULT: Binding to isolated DNA was most effective in aminoplatin, followed by methylplatin and oxiplatin. It differed by factors I. I, 0.35, 0.23, from the DNA binding of CDDP (factor I) for the three compounds, respectively. The cellular penetration, however, was fastest with oxiplatin (factor 10.1) followed by CDDP (1), aminoplatin (0.55), and methylplatin (0.27). The cytotoxicity of the three novel compounds followed the pattern of their cellular penetration with oxiplatin being the most active. U87-MG cells were resistant to CDDP, and in this cell line oxiplatin was more effective than CDDP in overcoming the resistance. CONCLUSION: This indicates cellular penetration to be an important feature and the ketogroup of oxiplatin to be beneficial in tricyclic platinum compounds. PMID- 11497296 TI - Inhibition by anticonvulsants of prostate-specific antigen and interleukin-6 secretion by human prostate cancer cells. AB - BACKGROUND: Anticonvulsants/antiepileptics inhibit high-frequency firing of action potentials that occur during an epileptic seizure, by mechanisms such as blocking sodium and calcium ion conductances. High sodium channel expression has been previously associated with aggressive behavior of prostate cancer (PCa) cell lines. The anticonvulsant phenytoin is known to inhibit the secretory activity and growth of human osteoblastic cells. MATERIALS AND METHODS: The effect of four anticonvulsants on the secretion of prostate-specific antigen (PSA) and interleukin-6 (IL-6) by human PCa cell lines (LNCaP, DU-145 and PC-3) was studied using immunoanalysis. RESULTS: Phenytoin and carbamazepine, which inactivate voltage-gated sodium channels, inhibited the secretion of PSA by LNCaP and IL-6 by DU-145 and PC-3 cell lines. Valproate and ethosuximide, which block calcium channels, had somewhat lower antisecretory effects. The growth of the three PCa cell lines in Matrigel was inhibited by all four anticonvulsants, at clinically relevant doses. CONCLUSION: Further study of the usefulness of anticonvulsants in PCa therapy is warranted. PMID- 11497297 TI - Modulation of IgG and complement receptor expression of phagocytes in kidney cancer patients during treatment with interferon-alpha. AB - BACKGROUND: The mode of action of interferon involves both direct cytotoxic and antiproliferative effects on the tumour cell and indirect effects that facilitate immune detection by the host. Among the immunological effects of interferon-alpha is the activation of monocytes. As opsonin receptors are crucial in the function of phagocytes, e.g. monocytes and neutrophils, their modulation by interferon alpha (INF-alpha) merit to be further clarified. We hypothesised that the role of phagocytes in defence against cancer is reflected in the expression of opsonin receptors for IgG and complement, which further could be modified by INF-alpha. PATIENTS AND METHODS: The expression of the receptors for IgG and complement was studied in neutrophils and monocytes from blood samples of 18 kidney cancer patients treated with INF-alpha and from 39 healthy individuals. Blood samples were collected prior/to and during treatment with INF-alpha, 4.5 to 13.5 MU t.d.w., subcutaneously. After lysing the red blood cells, the samples were incubated with fluorochrome conjugated monoclonal antibodies specific for IgG (Fc gammaRI, -RII and -RIII) and complement (CR1, CR3) receptors and then analysed in flow cytometry. The results were given as the mean log fluorescence intensity (a measure of receptor number) and as the proportion of receptor-positive cells. In the in vitro experiments, the direct effect of interferon-alpha on the receptors of neutrophils and monocytes was studied. RESULTS: In patients before any treatment, the expression of CR3 and Fc gammaRI receptors in neutrophils and all receptors except Fc gammaRIII in monocytes was significantly raised when compared to the controls. Treatment with INF-alpha, induced statistically significant; transient changes in CR1-receptor expression in neutrophils and Fc gammaRI expression in monocytes. Incubation of blood cells with INF-alpha in vitro confirmed the induction of CR1 receptors in neutrophils. CONCLUSION: Changes in receptor expression reflect the inflammatory activation of phagocytes in metastatic kidney cancer. The pattern of receptor expression differs from that observed in infectious diseases. Interferon-alpha both in vivo and in vitro modulates the expression of phagocytic receptors. PMID- 11497298 TI - Antitumour activity of N6-substituted PMEDAP derivatives against T-cell lymphoma. AB - The antitumour activity of four N6-substituted PMEDAP derivatives, Me2NEt-PMEDAP, allyl-PMEDAP, Me2-PMEDAP and cypr-PMEDAP, selected on the basis of their in vitro cytostatic activity, was studied in an in vivo model of haematological malignancy of inbred Sprague-Dawley rats. These compounds are believed to serve as the prodrugs of another (phosphonomethoxy)ethyl derivative, PMEG (9-[2 phosphonomethoxy) ethyl] guanine. We compared their toxicity and ability to inhibit tumour development in two different dosage regimes with those of their parent compound PMEDAP, as well with PMEG. The study confirmed the anticancer efficacy of the parental compound PMEDAP. Unlike PMEDAP, its N6-mono- and disubstituted congeners Me2NEt-PMEDAP, allyl-PMEDAP and Me2-PMEDAP were less potent or exhibited the same antineoplastic effect as PMEDAP. cypr-PMEDAP significantly decreased the survival of lymphoma-bearing rats due to high toxicity, which was approximately the same as that of PMEG. Therefore, these acyclic nucleoside phosphonates substituted at the 6-position of 2,6 diaminopurine ring do not seem to be promising drugs for the treatment of haematological malignancies. PMID- 11497299 TI - Transfection with mdm2-antisense or wtp53 results in radiosensitization and an increased apoptosis of a soft tissue sarcoma cell line. AB - Soft tissue sarcomas (STS) are mostly resistant after radiation treatment and are characterized by a rather low rate of apoptosis. The aim of this study was to test, in the p53 mutant STS cell line US8-93, the effect of a combined treatment with DNA transfection--either with mdm2 antisense oligodesoxynucleotides (mdm2 AS) or with a wild-type p53-plasmid (wtp53)--and the effects of irradiation on radiosensitivity. Mdm2-sense oligodesoxynucleotides (mdm2-SE) and a GFP-plasmid (GFP) were applied as controls. In order to evaluate the treatment radiation sensitization (clonogenic survival), apoptotic cell death and P53/MDM2-protein expression were determined. A moderately increased radiation sensitization was observed when comparing clonogenic survival after 2 Gy irradiation between cells transfected either with the control mdm2-SE (48%) or with mdm-2 AS (30%). At the same irradiation dose, clonogenic survival of wtp53-plasmid transfected cells (32%) was about 2-fold less than in the cells transfected with the control GFP plasmid (61%). This enhancement factor of radiation sensitization was increased by about 3-fold at 4 Gy irradiation. Furthermore, an increase in apoptotic cells was already detectable by up to 7.7% (mdm2-AS) in comparison to 3.1% (mdm2-SE control) 72 hours after transfection. In parallel, the percentage of apoptotic cells could be further elevated after subsequent irradiation with 12 Gy by up to 15% (mdm2-AS) compared to 5.7% (mdm2-SE control). A striking result was obtained with the combined treatment of a wtp53 and 12 Gy irradiation which produced in 25% and 38.9% of apoptotic cells 48 hours and 72 hours after transfection, respectively. We can therefore conclude that the sensitivity of radiation therapy is enhanced by DNA transfection with wtp53 or mdm-2 AS ODNs for the correction of the p53-mdm2 balance in STS in vitro. PMID- 11497300 TI - Effect of Porphyridium sp. polysaccharide on malignant cell transformation by Moloney murine sarcoma virus. AB - The polysaccharide produced by the red microalga Porphyridium sp. was highly inhibitory for cell transformation of mouse fibroblast cells by an MSV-124 virus stock. This inhibition was most effective if the polysaccharide was added 2 h before or at the time of infection. The finding that the inhibition of cell transformation by MuSV-124 was reversible after removal of the polysaccharide suggested that Porphyridium sp. polysaccharide inhibited a late step after provirus integration into the host genome. Addition of the polysaccharide post infection significantly reduced the number of transformed cells, but its effect was less marked than that obtained when the polysaccharide was added before or at the time of infection. These findings support the possibility that the polysaccharide may affect early steps in virus replication cycle, such as virus absorption into the host cells, in addition to its effect on a late step after provirus integration. PMID- 11497301 TI - The response of human breast tumour cell lines with altered polymerase beta levels to cisplatin and radiation. AB - MCF 7 (human breast carcinoma cells) and mutants transfected with the DNA polymerase beta gene were tested for response to cisplatin, radiation and combined treatments. The transfected cells showed a higher level of polymerase beta activity and were more resistant to radiation and cisplatin compared to the parental cell line. Further studies showed that for isosurvival treatments the mutant cells were more effective in sublethal radiation damage repair compared to the parental line. The combination of cisplatin with radiation showed effective radiosensitization which was less in the mutants compared to the parental line. In addition, the sequence of cisplatin before irradiation was more effective then cisplabn after irradiation. Pre-exposure to low levels of cisplatin for up to 24 h before irradiation showed a small significant adaptive response in one mutant line at 8 h and while similar trends were observed in the parental lines at earlier times they were not significant. In summary our data show that polymerase beta and thus base excision repair may play a role in cellular responses to cisplatin and radiation. PMID- 11497303 TI - The first 100 days. PMID- 11497302 TI - Overexpression of peroxiredoxin in human breast cancer. AB - The peroxiredoxins (Prx) are a family of 25 kDa peroxidases that can reduce H2O2 using an electron from thioredoxin (Trx) or other substances. The mammalian Prx family is divided into six groups (Prx I-VI) on the basis of homology of amino acid sequences. They are located in the cytosol and play a role in the cell signaling system. Previous reports have shown that Prx II has proliferative and anti-apoptotic properties and thus may induce carcinogenic changes. We conducted this study to reveal the change in expression of Prx in human breast cancer in comparison to normal tissues. Western immunoblotting using Prx type I, II and III antibodies was undertaken on 24 human breast cancer tissues and normal counterparts. We used antibodies against purified recombinant NKEF-A/PAG, NKEF-B and MER 5 which are the Prx isoforms. Type I Prx was overexpressed in the cancer tissues of 21 patients (87.5%), type II in 18 patients (75%) and type III in 19 patients (79.2%) in relation to normal tissue. However, no significant relationship was found between Prx overexpression and clinicopathological parameters of breast cancer such as tumor size, lymphatic invasiveness, hormone receptor status or nuclear and histologic grade. In conclusion, Prx is overexpressed in breast cancer tissues to a great extent suggesting that Prx has a proliferative effect and may be related to cancer development or progression. PMID- 11497304 TI - Trends in the consumer price index. PMID- 11497305 TI - Hospitalist programs: are they good for your practice? AB - With the changes brought about in health care delivery by the growth of Managed Care, the attractions of hospitalist programs are becoming increasingly evident. As the programs have grown in usage, and program models have evolved from mandatory to voluntary, primary care physicians (PCPs) and independent practice organizations (IPAs) have begun to recognize their positive impact on their practices, from both financial and logistic perspectives, as well from those of patient care. This article discusses that effect and reviews the procedures and ramifications of one of the newer program models. Quality of patient care and the provision of successful outcomes via clinical guidelines are addressed as indications of successful hospitalist programs. PMID- 11497306 TI - How to handle part-time, flex-time, and job-sharing employees. AB - Offering employment structures other than traditional full-time positions in your practice can help you draw excellent job applicants and also can enable you to increase morale, job satisfaction, and productivity. However, there are many decisions you must make when offering a part-time, flex-time, or job-sharing position. This article explores the pros and cons of offering alternative job structures. It suggests ways to make part-time, flex-time, or job-sharing positions work most effectively, both for the employee and for your practice. In addition, this article suggests which positions are best suited to alternative structures. PMID- 11497307 TI - Create an environment for employee satisfaction, loyalty, and commitment. PMID- 11497308 TI - Re-tooling your practice: analyze and fix your practice's problem areas. PMID- 11497309 TI - Is it safe? Security speed bumps on the information highway. AB - The Internet is not only changing the face of computing but is redefining the manner in which medical information is stored, manipulated, and disseminated. However, before patients and clinicians can come to trust the Internet as much as traditional paper medical records, the issue of data security must be adequately addressed. Every clinician who works with patient data that are potentially accessible through the Internet should be familiar with the relevant security technologies, ranging from encryption and firewalls to intrusion detection and anti-virus software. Clinicians should also realize that security is a process and that the majority of security violations are simply the result of careless patient information handling and inadequate policies. PMID- 11497310 TI - Interactive patient education: the X-Plain model. AB - Responding to competitive pressure to enhance services and patient satisfaction while reducing cost, many healthcare organizations are turning to interactive health communication to enhance their practices. This article reviews computer based patient education by focusing on a product, X-Plain, and describing how it is used in medical practices. The following benefits of computer-based patient education are discussed: risk management, time saving, enhanced patient satisfactions, and meeting regulatory requirements. Two classes of computer-based patient education systems are distinguished: documentation systems used in the back office to inform patients about their diagnoses and treatments and to document patient education; and public kiosks used in the reception area to provide general information, promote the practice's services, and collect survey data. The advantages of delivering patient education through the Web such as global networking and presentation of customized health information are discussed. PMID- 11497311 TI - Physician recruitment websites: the territory ahead. AB - Recruitment is no longer just a face-to-face interaction between a recruiter and a job-seeker. To remain up-to-date with current technology, physicians should use the various electronic tools available today, including recruitment websites, video previews, and PC-to-PC interviews to get the job they've been looking for. This article discusses the pros and cons of these interfaces. PMID- 11497312 TI - Safety and compliance-related hazards in the medical practice: Part one. AB - Safety and risk management hazards are a fact of life for the medical practice, and the costs of these incidents can place the group at significant risk of liability. Good compliance and risk management programs help minimize these incidents, improve staff morale, increase a practice's visibility in the community, and positively affect the practice's financial and operational bottom line performance. Medical practices that implement effective safety and risk management programs can realize savings in staffing costs, operational efficiency, morale, insurance premiums, and improved third-party relationships while at the same time avoiding embarrassing risks, fines, and liability. This article outlines some of the most common safety and risk management-related deficiencies seen in medical practices today. The author explains how to remedy these deficiencies and provides a self-test tool to enable the reader to assess areas within his or her own practice in need of attention. PMID- 11497314 TI - Confidentiality of peer review. PMID- 11497313 TI - Legal update: a roadmap of the recent changes to the medico-legal landscape. AB - This article provides an overview of recent changes to federal law that physicians should be aware of The Final Compliance Program Guidance for Individual and Small Group Practices, the HIPAA privacy regulations, and the Fraud and Abuse Safe Harbors are discussed specifically. Also addressed are miscellaneous tax changes and the lapse of the Advisory Opinions. PMID- 11497315 TI - Identifying, assessing and managing high-risk patients under Medicare risk contracts. AB - Physicians participating in Medicare managed care plans are increasingly sharing in the financial risks of providing care to enrollees. Although capitation payments are calculated to reflect the average costs for an entire patient panel, there is typically great variation among patients, from healthy individuals to patients requiring high volume and high-cost care. To be successful, physicians must identify high-risk seniors, develop care plans to maintain health and functioning, and monitor health status and ongoing care and treatment. Techniques for performing a comprehensive geriatric assessment include written questionnaires and interviews during office appointments. The goal is to evaluate each patient's health status, medical risk factors, psychosocial profile, cognitive capacities, and functional capabilities and limitations. After frail and at-risk seniors are identified, the physician must collaborate with the case manager, geriatric nurse practitioner, other caregivers, and the patient and family members to initiate and maintain effective care management. PMID- 11497317 TI - Costs and savings to practices under HIPAA. Health Insurance Portability and Accountability Act. PMID- 11497316 TI - What physicians should know about documenting disability insurance claims. AB - Physicians are frequently called upon to provide opinions to insurers or to the Social Security Administration regarding disability issues. This article discusses various issues that frequently arise and offers suggestions on how to assist patients in obtaining needed benefits. PMID- 11497318 TI - Economic profitability: 1999-2001. PMID- 11497319 TI - Schedules of reinforcement as determinants of human causality judgments and response rates. AB - Experiments examined the effect of relationships between a response and an outcome on human judgments of causal effectiveness. In Experiment 1, the time between outcomes obtained on a variable ratio (VR) schedule became the intervals for a yoked variable interval (VI) schedule. Response rates were higher on the VR than on the VI schedule. In Experiment 2, the number of responses required per outcome on a VR schedule were matched to that on a master VI 20-s schedule. Both ratings of causal effectiveness and response rates were higher in the VR schedule. In Experiment 3, tandem VI fixed-ratio (FR) schedules produced higher rates and judgments than equivalent conjunctive VI FR schedule. In Experiment 4, a VI schedule with a reinforcement requirement for a short interresponse time (IRT) produced higher rates and judgments than a simple VI schedule. These results corroborate the view that schedules are a determinant of both response rates and causal judgments. Few current theories of causal judgment predict this pattern of results. PMID- 11497320 TI - Transfer between contextual conditional discriminations: an examination of how stimulus conjuctions are represented. AB - In 2 experiments, when rats were placed in 1 pair of contexts, A and B, 2 relationships were in force (X --> food and Y --> no food), and when they were placed in another pair of contexts, C and D, the complementary relationships were operative (Y --> food and X --> no food). In Experiment 1, rats then received a 2nd discrimination that was either contextually congruent (in A and B, Y --> food and X --> no food; in C and D, X --> food and Y --> no food) or contextually incongruent (in A and D, Y --> food and X --> no food; in C and B, X --> food and Y --> no food) with the 1st discrimination. In Experiment 2, the 1st discrimination, involving X and Y, was interleaved with a 2nd discrimination, involving V and W, that was again either congruent (in A and B, V --> food and W -> no food) or incongruent (in A and D, V --> food and W --> no food) with the 1st discrimination. The congruent discriminations were acquired more readily than were the incongruent discriminations. PMID- 11497321 TI - Discrimination of structure: I. Implications for connectionist theories of discrimination learning. AB - In each of 4 experiments animals were given a structural discrimination task that involved visual patterns composed of identical features, but the spatial relations among the features were different for reinforced and nonreinforced trials. In Experiment 1 the stimuli were pairs of colored circles, and pigeons were required to discriminate between patterns that were the mirror image of each other. A related task was given to rats in Experiment 2. Subjects solved these discriminations. For Experiment 3, some pigeons were given a discrimination similar to that used in Experiment 1, which they solved, whereas others received a comparable task but with 3 colored circles present on every trial, which they failed to solve. The findings from Experiment 3 were replicated in Experiment 4 using different patterns. The results are difficult to explain by certain connectionist theories of discrimination learning, unless they are modified to take account of the way in which compound stimuli are structured. PMID- 11497322 TI - Shock-induced hyperalgesia: IV. Generality. AB - Brief-moderate shock (3, 0.75 s, 1.0 mA) has opposite effects on different measures of pain, inducing antinociception on the tail-flick test while lowering vocalization thresholds to shock and heat (hyperalgesia) and enhancing fear conditioned by a gridshock unconditioned stimulus (US). This study examined the generality of shock-induced hyperalgesia under a range of conditions and explored parallels to sensitized startle. Reduced vocalization thresholds to shock and antinociception emerged at a similar shock intensity. Severe shocks (3, 25 s, 1.0 mA or 3, 2 s, 3.0 mA) lowered vocalization threshold to shock but increased vocalization and motor thresholds to heat and undermined fear conditioned by a gridshock or a startling tone US. All shock schedules facilitated startle, but only brief-moderate shock inflated fear conditioning. The findings suggest that brief-moderate shock enhances the affective impact of aversive stimuli, whereas severe shocks attenuate pain. PMID- 11497323 TI - Assessing the contributions of S-O and R-O associations to differential-outcome matching through outcome reversals. AB - Pigeons were trained on symbolic matching with 2 samples, 2 pairs of comparisons, and different outcomes for the correct responses within each comparison pair. For one group, the 2 samples were also associated with different outcomes, whereas for another group, they were not. When the response-outcome (R-O) relations for one pair were subsequently reversed, the group trained with differential sample outcome (S-O) associations was significantly disrupted in its performance on both reversed- and nonreversed-outcome trials. By contrast, the group trained with just differential R-O associations was disrupted only on reversed-outcome trials. These results were replicated when the outcomes on the initially nonreversed trials were then reversed. The findings indicate that differential S-O associations, when present, have a stronger influence on matching performances than differential R-O associations. They are also consistent with hierarchical and configural models of discriminative control. PMID- 11497324 TI - Superordinate categorization via learned stimulus equivalence: quantity of reinforcement, hedonic value, and the nature of the mediator. AB - Three experiments examined superordinate categorization via stimulus equivalence training in pigeons. Experiment 1 established superordinate categories by association with a common number of food pellet reinforcers, plus it established generalization to novel photographic stimuli. Experiment 2 documented generalization of choice responding from stimuli signaling different numbers of food pellets to stimuli signaling different delays to food reinforcement. Experiment 3 indicated that different numbers of food pellets did not substitute as discriminative stimuli for the photographic stimuli with which the food pellets had been paired. The collective results suggest that the effective mediator of superordinate categories that are established via learned stimulus equivalence is not likely to be an accurate representation of the reinforcer, neither is it likely to be a distinctive response that is made to the discriminative stimulus. Motivational or emotional mediation is a more likely account. PMID- 11497325 TI - Topography of spatially directed conditioned responding: effects of context and trial duration. AB - Domesticated quail (Coturnix japonica) received a discrete conditioned stimulus (CS) at one end of the experimental chamber paired with the opportunity to copulate with a female quail (the unconditioned stimulus) in a goal box located 112 cm away. Approach to the CS (sign tracking) and approach to the goal area (goal tracking) were measured. The duration of exposure to the experimental context (C) was varied in Experiment 1, and the duration of the conditioning trials (T) was varied in Experiment 2 for independent groups, creating C/T ratios of 1.0, 1.5, 4.5, 45, and 180. Contrary to previous reports of a direct relation between the C/T ratio and conditioned responding, in the present experiments, a shift in the topography and stimulus control of conditioned behavior occurred. Low C/T ratios (1.0-4.5) produced goal tracking controlled by contextual cues, whereas high C/T ratios (45 and 180) produced sign tracking controlled by the discrete CS. PMID- 11497326 TI - Sequential and simultaneous feature positive discriminations: occasion setting and configural learning in human Pavlovian conditioning. AB - Using a conditioned suppression preparation, the authors investigated sequential (X --> A+/A-) versus simultaneous (XA+/A-) feature positive (FP) discrimination learning in humans. The sequential discrimination was expected to be resolved by means of a Feature X Modulated Target A-US association and the simultaneous discrimination by a feature X-US association. After sequential FP training, extinction of Feature X did not affect discriminative X --> A/A responding (Experiment 1), and X transferred its modulatory ability only to new targets, B, that had also been modulated (Experiment 2). This suggests that the sequential FP discrimination indeed resulted in occasion setting. Unlike expected, Feature X Extinction did not affect discriminative XA/A responding after simultaneous FP training (Experiment 3), while at the same time Feature X did show the predicted nonselective transfer to new targets, B (Experiment 4). J. M. Pearce's (1987) configural learning theory can account for most but not all findings of Experiments 3 and 4. PMID- 11497327 TI - Thermal and physical properties of bakery products. AB - This article reviews the measurement techniques, prediction models, and data on thermo-physical properties of bakery products: specific heat, thermal conductivity, thermal diffusivity, and density. Over the last decade, investigation has focused more on thermo-physical properties of nonbread bakery products. Both commonly used and new measurement techniques for thermo-physical properties reported in the publication are presented with directions for their proper use. Data and prediction models are tabulated for the range of moisture content and temperature of the bakery products. PMID- 11497328 TI - Inulin and oligofructose as dietary fiber: a review of the evidence. AB - This critical review article examines the composition and source of inulin and oligofructose, the physiological effects of their consumption, and how these materials relate to the concept of dietary fiber. Inulin and oligofructose are fructans extracted on a commercial basis from the chicory root. Inulin has been defined as a polydisperse carbohydrate material consisting mainly, if not exclusively, of beta (2-1) fructosyl-fructose links ranging from 2 to 60 units long. Native chicory inulin has an average degree of polymerization (DP) of 10 to 20, whereas oligofructose contains chains of DP 2 to 10, with an average DP of 4. While a universally accepted definition for dietary fiber does not exist, it is generally agreed that this term includes saccharides (+ lignin) that are not hydrolyzed or absorbed in the upper part of the gastrointestinal tract. These materials reach the colon, where they may be totally fermented, partially fermented, or remain unfermented. In addition, fibers contribute to fecal bulking. Inulin and oligofructose are not digested in the upper part of the gastrointestinal tract or are they absorbed and metabolized in the glycolytic pathway, or directly stored as glycogen like 'sugars' or starches. None of the molecules of fructose and glucose that form inulin and oligofructose appear in the portal blood. These materials are quantitatively fermented by the microflora of the colon; further, it has been demonstrated that this fermentation leads to the selective stimulation of the growth of the bifidobacteria population. After reviewing their chemistry, origin, and physiological effects, it is the opinion of the authors that inulin and oligofructose are dietary fiber. They share the basic common characteristics of dietary fibers, that is, saccharides of plant origin, resistance to digestion and absorption in the small intestine, and fermentation in the colon to produce short-chain fatty acids that are absorbed and metabolized in various parts of the body. Moreover, this fermentation induces a bulking effect. PMID- 11497329 TI - The effect of ingesting olestra-based foods on feeding behavior and energy balance in humans. AB - This article reviews currently published works on the effects of olestra on appetite and energy intake (EI) in humans. To date 24 studies have examined the impact of olestra-containing foods on aspects of feeding behavior, which are published in 20 reports. The general response of human subjects to olestra-based decreases in dietary energy density (ED) is either poor caloric compensation or partial (nonmacronutrient specific) increases in EI. Averaging the degree of energy compensation across 22 studies gives a value of 27% (nonweighted mean). In studies where compensation occurred, fat intake but not EI was reduced. These effects appear to occur in both lean and obese, men and women and under a variety of conditions ranging from the laboratory to real life. However, all but two of these studies were short term. One study suggests that in subjects to whom weight loss is desirable these deficits can persist for up to 3 months. Subjects with no wish to lose weight may compensate better over longer periods. In another 3-month study, ingestion of olestra-based foods did not induce energy deficits but limited the significant weight gain seen on a full-fat control. The longer term effects of olestra on body weight requires further investigation. There is evidence that restrained eaters tend to eat slightly more of olestra-based foods if they know that they are reduced in fat and energy. This is probably a general response to low-fat foods rather than to olestra per se. The fact that olestra based foods have the potential to provide the sensory qualities of real fat suggests that these foods may be particularly effective in habitual high-fat consumers with a sensory preference for dietary fat. PMID- 11497330 TI - Expression of hsp 27, hsp 60, hsc 70, and hsp 70 by immortalized human proximal tubule cells (HK-2) following exposure to heat shock, sodium arsenite, or cadmium chloride. AB - The expression of hsp 27, hsp 60, hsc 70, and hsp 70 mRNA and protein was determined in immortalized human proximal tubule cells (HK-2) exposed to heat shock, sodium arsenite, or cadmium chloride (CdCl2) under both acute and extended conditions of exposure. It was demonstrated that the HK-2 cells did not exhibit the classic heat-shock response when subjected to an acute physical (heat) or chemical stress (sodium arsenite or CdCl2). Heat stress, elevated temperature at 42.5 degrees C for 1 h, caused a marked increase only in hsp 70 mRNA and protein, but not hsp 27 or hsp 60 mRNA and protein. Similar results were obtained when the cells were subjected to a classic chemical stress of exposure to 100 microM sodium arsenite for 4 h or CdCl2 for 4 h. These findings were in contrast to those found previously with mortal human proximal tubule (HPT) cells, where acute stress by all three stimuli elicited marked increases in hsp 27, hsp 60, and hsp 70 mRNA and protein. It was shown that the basal levels of expression of hsp 27 and hsp 60 in the HK-2 cells were elevated when compared to those found in unstressed HPT cells and that the basal levels were similar to those found in HPT cells under stress conditions. These results suggest that the failure of the HK-2 cells to increase hsp 27 and hsp 60 levels in response to physical and chemical stress is because they already possess elevated basal levels of these proteins. This would indicate that one or more of the genetic events that resulted in the immortalization of the HK-2 cells also elicited a stress response for hsp 27 and hsp 60, but not for hsp 70, stress response family members. Overall, the results suggest that although there are differences in the regulation of the stress response between the immortal HK-2 and mortal HPT cell lines, as long as these differences are recognized, the HK-2 cell line should be a valuable adjunct to study the stress response of the proximal tubule in general and when exposed to environmental pollutants such as cadmium. PMID- 11497331 TI - Biomarker and bioaccumulation responses of Asian clams exposed to aqueous cadmium. AB - Measured responses of biochemical or physiological indicators have been suggested to reflect thresholds where pollutants exert their initial effect. Responses in cellulolytic enzyme activity and DNA strand breakage of the Asian clam Corbicula fluminea exposed to cadmium in the laboratory were measured and metal body burdens were determined concurrently. Clams were exposed to aqueous cadmium concentrations of 3, 6, 12, or 25 ppb for 23 and 28 d. Cadmium concentrations in clam tissue were highest in lower cadmium treatments, and body burdens increased with length of exposure in only the 28-d experiment. Cellulolytic enzyme activity decreased with increasing cadmium concentrations for clams in the 28-d experiment. Induced enzyme activities were observed in cadmium treatments for both experiments and are thought to precede declines in activity through the length of exposure. Significant reductions in DNA strand lengths of cadmium exposed clams were observed by wk 3 in the 23-d exposure and by wk 2 in the 28-d exposure. Reduced DNA strand lengths in these cadmium treatments for the 28-d exposure precede significant declines in cellulolytic activity at subsequent sampling events. Combining these data with observations of mortality in higher cadmium treatments suggests that impairment of DNA structural integrity and reduced digestive enzyme activity may indicate metal-induced stress in clams. PMID- 11497332 TI - Mortality in mice infected with an amyocarditic coxsackievirus and given a subacute dose of mercuric chloride. AB - An amyocarditic strain of coxsackievirus B3 (CVB3/0) induces heart damage when inoculated into selenium (Se)-deficient mice. Mercury (Hg), an Se antagonist, is known to aggravate viral infections. The experiments reported here assessed the effect of prior Hg treatment in mice subsequently inoculated with an amyocarditic strain of coxsackievirus. A pilot study showed that under our conditions the maximum tolerated dose of HgCl2 in uninfected mice was 6 mg HgCl2/kg body weight. In the main study, doses of 0, 3 or 6 mg HgCl2/kg body weight were administered intraperitoneally (ip) to 7-wk-old male mice fed a standard chow diet. Two hours later, half the mice were inoculated ip with CVB3/0. Ten days postinoculation, no mortality was observed in mice given only virus. In mice not given virus, 10% injected with 6 mg HgCl2/kg body weight died. On the other hand, 64% of the mice given both virus and 6 mg HgCl2/kg body weight died. Fifteen percent of the hearts from virus-infected mice given 3 mg HgCl2/kg body weight and 33% of the hearts from virus-infected mice given 6 mg HgCl2/kg body weight exhibited a higher incidence of lesions than hearts from mice-given virus alone. Moreover, viral heart titers were elevated in infected mice injected with 6 mg HgCl2/kg body weight compared to infected mice receiving no Hg. Thus, an amyocarditic coxsackievirus given to mice after a nonlethal subacute dose of Hg results in mortality, increased incidence of heart lesions, and elevated viral heart titers. These results demonstrate the important role of toxic elements in determining the severity of viral infections. PMID- 11497333 TI - Metabolism of aflatoxin B1 by normal human bronchial epithelial cells. AB - Aflatoxin B, (AFB1) is a potent hepatocarcinogen in animal models and a suspected carcinogen in humans. High concentrations of AFB, have been found in respirable grain dusts, and may therefore be a risk factor for human lung cancer in certain occupations. To study the potential for AFB, activation in human lung, cytochrome P-450 (CYP)-mediated activation and glutathione S-transferase (GST)-mediated detoxification of AFB1 were examined in cultured normal human bronchial epithelial (NHBE) cells. Cells were exposed to 0. 15 microM or 1.5 microM AFB, for 48 h and media was collected for metabolite analysis by high-performance liquid chromatography (HPLC). At 0. 15 microM, AFB1 was metabolized only to the detoxified metabolite aflatoxin Q1 (AFQ1). At 1.5 microM AFB1, both aflatoxin M1 (AFM1), and AFQ1 were produced. Cells pretreated with 50 degrees M 3 methylcholanthrene (3MC), a CYP 1A inducer, for 72 h prior to 0.15 microM AFB1, produced the activated AFB1 8,9-epoxide (AFBO). Similarly, microsomes prepared from 3MC-pretreated cells formed AFBO, but microsomes from noninduced cells did not. While AFB1-DNA adducts were not detected at low AFB1 concentrations in untreated NHBE, 3MC induction caused the production of AFB1-DNA adducts at 0.015 and 0.15 microM AFB1. Western immunoblots showed that the primary CYP isoforms responsible for AFB1 activation in the liver, 1A and 3A4, to be constitutively expressed in NHBE cells. Expression of CYP 1A was significantly increased in 3MC pretreated cells, while CYP 3A4 expression increased slightly, but not to the extent of the 1A isoforms. The principal AFBO detoxifying enzyme, glutathione S transferase (GST), was constitutively expressed in NHBE cells, and was increased approximately twofold by 3MC pretreatment. Cytosolic fractions from neither control nor 3MC-induced NHBE had measurable AFBO conjugating activity, indicating that these cells may lack AFB1-relevant GST activity. From these data, it appears that NHBE cells activate AFB1 inefficiently, but possess CYPs reportedly responsible for metabolism of AFB1. These data support earlier findings showing modest CYP-mediated AFB1 activation in human airways, but indicate that exposure to polycyclic aromatic hydrocarbons (PAHs), such as 3MC, which induce CYP(s) that specifically activate AFB1 may increase the harmful effects of AFB1 exposures in human airways. PMID- 11497334 TI - Metabolism of the styrene metabolite 4-vinylphenol by rat and mouse liver and lung. AB - Styrene is a widely used chemical in the reinforced plastics industry and in polystyrene production. Its primary metabolic pathway to styrene oxide and then to styrene glycol, which is further metabolized to mandelic acid and phenylglyoxylic acid, has been well studied. However, a few studies have reported finding a minor metabolite, 4-vinylphenol (4-VP), in rat and human urine. The present studies sought to determine if the formation and metabolism of 4-VP in rat and mouse liver and lung preparations could be measured. When styrene was incubated with hepatic and pulmonary microsomal preparations, 4-VP formation could not be measured in these preparations. However, considerable 4-VP metabolizing activity, as determined by the loss of 4-VP, was observed in both mouse and rat liver and lung microsomal preparations. 4-Vinylphenol metabolizing activity in mouse liver microsomes was three times greater than that in rat liver microsomes, and activity in mouse lung microsomes was eight times greater than that in rat lung microsomes. This activity was completely absent in the absence of NADPH. Studies with cytochrome P-450 inhibitors indicated the involvement of CYP2E1 and CYP2F2. Induction of CYP2E1 by pyridine resulted in an increase in 4 VP metabolism by mouse hepatic microsomes but not by pulmonary microsomes. The metabolite(s) formed as a result of this oxidative pathway remain to be identified. In additional studies, glutathione conjugation appeared to be involved in 4-VP metabolism with the highest activity being in mouse lung, with or without the addition of NADPH. PMID- 11497335 TI - Organising evidence on QT prolongation and occurrence of Torsades de Pointes with non-antiarrhythmic drugs: a call for consensus. AB - BACKGROUND: The growing list of non-antiarrhythmic drugs associated with QT prolongation and the relevant regulatory interventions have generated concern for two reasons. First, QT prolongation is sometimes viewed as an intrinsic effect of a whole therapeutic class (for example, antihistamines), whereas, in many cases, it is displayed only by some compounds within a given class of non-antiarrhythmic drugs because of an effect on cardiac repolarisation. Second, drug-induced Torsades de Pointes are still considered idiosyncratic, totally unpredictable adverse drug reactions, whereas a number of risk factors for their occurrence is now recognised. OBJECTIVES: In order to increase awareness among prescribing physicians that many non-antiarrhythmic drugs can affect cardiac repolarisation, we would like to propose a comprehensive and updated list of QT-prolonging drugs that should be a starting point to maintain a "consensus list" to be periodically updated. METHODS: The drug list was generated by performing a Medline search, by using published lists as starting points to retrieve the relevant references quoted in each article and by considering the International Registry for Drug induced Arrhythmias maintained by the Georgetown University and mainly based on the FDA approved labelling. RESULTS: The drug list presented in this paper: (1) includes virtually all non-antiarrhythmic drugs with QT-prolonging potential, (2) organises the available information on each drug at different levels of clinical relevance and (3) is as up-to-date as possible in order to provide a fast track for the clinical pharmacologist to retrieve the original publications. CONCLUSIONS: This list should be considered as a starting point to call for consensus on: (1) the criteria used to generate the list, (2) possible ways to implement the use of this list as a quick reference for clinicians, for instance by providing a "proarrhythmic score" for each drug, and (3) inclusion/exclusion of a given agent into the list on the basis of evidence that may not be available to us. PMID- 11497336 TI - Microdialysis. A novel tool for clinical studies of anti-infective agents. AB - In vivo microdialysis (MD) is an innovative clinical technique that has been employed in preclinical research and metabolic studies in patients for more than a decade. Recently, MD has been adopted for human drug studies and has opened up the opportunity to quantify tissue drug distribution in vivo. The particular advantage of MD for the anti-infective field relates to the fact that MD allows for online measurement of the unbound, pharmacologically active drug fraction in the interstitial space fluid (ISF), the anatomically defined target site for most bacterial infections. The aim of this review is to provide an overview of the current literature about MD in anti-microbial drug studies. It will be shown that MD has become feasible in most human tissues including brain and lung. So far, several MD studies have demonstrated that anti-microbial concentrations at the effect site may be subinhibitory, although effective concentrations are attained in serum, a finding that has significant impact on clinical decision making. In addition to its property as a pharmacokinetic sampling technique, MD offers unique opportunities in pharmacokinetic-pharmacodynamic (PK-PD) research and has the potential to streamline the decision process on proper drug dosing in drug development. PMID- 11497337 TI - The quality of public and private pharmacy practices. A cross sectional study in the Savannakhet province, Lao PDR. AB - AIM: The aim was to study the practices of public and private pharmacies in the Savannakhet province, Lao PDR, in relation to defined aspects of good pharmacy practice (GPP) and rational use of drugs (RUD). STUDY DESIGN: The study design was cross-sectional using structured interviews and observations. A total of 105 drug sellers (31% and 4% had pharmacy education at public and private pharmacies, respectively) were interviewed, and the pharmacies were inspected. In addition, 576 customers were interviewed immediately after the drug transaction and all their drug purchases were recorded. Facility, patient-care and prescribing indicators covering aspects of GPP and RUD were used to measure and compare the quality of services of the pharmacies. RESULTS: The results showed that public pharmacies differed significantly from private pharmacies, with lower mean scores for availability of essential drugs (5.1 vs 6.4), 95% confidence interval (CI 2.23, -0.34) and essential materials (5.6 vs 6.9, 95% CI -2.40, -0.20), and with a higher percentage of antibiotics dispensed (34% vs 24%, P<0.02), as well as more injections (31% vs 7%, P < 0.001) and drugs per customer (2.4 vs 1.4, 95% CI 0.84, 1.16). More drug purchases were decided by health workers at public pharmacies than at private pharmacies (92% vs 16%). At public pharmacies, significantly more drugs were prescribed from the National Essential Drug List (76% vs 56%, P=0.004), and more drugs had an international non-proprietary name (67% vs 35%, P<0.001). There was no significant difference regarding order in the pharmacy, oral information and drugs adequately labelled at the public pharmacies compared with the private pharmacies. CONCLUSION: In spite of the differences shown, both public and private pharmacies performed suboptimally in relation to several aspects of GPP and RUD. The lack of essential drugs, essential materials, information on drug use and adequate drug labelling all contribute to irrational use of drugs. Interventions are needed to improve practice and drug use. PMID- 11497338 TI - Study of the drug-drug interaction between simvastatin and cisapride in man. AB - OBJECTIVE: The objective of our study was to evaluate in humans the drug-drug interaction occurring during the concomitant administration of cisapride and simvastatin, two well-known substrates of CYP3A4. METHODS: Eleven healthy men aged between 20 years and 35 years gave their written informed consent to participate in the study. Each participant received repeated doses of cisapride and/or simvastatin. At first, subjects received cisapride alone, 10 mg every 8 h, for 3 days. Then, the drug was given at the same regimen during concomitant administration of simvastatin, 20 mg every 12 h for 4 days, starting on the night of day 3. Finally, cisapride was stopped and subjects received simvastatin (20 mg every 12 h) for four additional days. RESULTS: Simvastatin administration caused a 14 +/- 20% increase in the AUC0-8 of cisapride. In contrast, plasma concentrations of simvastatin were unaltered by the coadministration of cisapride, whereas plasma concentrations of simvastatin acid, its active metabolite, were decreased by 33 +/- 24%. CONCLUSION: The concomitant administration of the prokinetic agent cisapride and the 3-hydroxy-3 methylgluaryl CoA reductase inhibitor simvastatin resulted in altered pharmacokinetics of both drugs. Increased plasma concentrations of cisapride suggest that some patients may be at risk of toxicity while receiving both drugs, whereas the decrease in simvastatin acid plasma concentrations suggests that cholesterol lowering effects of simvastatin treatment may be blunted. PMID- 11497339 TI - Pharmacokinetics of viral antibodies after administration of intravenous immunoglobulin in patients with chronic lymphocytic leukaemia or multiple myeloma. AB - OBJECTIVE: Intravenous immunoglobulin (IVIG) preparations are derived from human pooled plasma and should fulfil high standards of purity and viral safety. Introduction of additional purification steps, however, may result in modulation of the biological properties of immunoglobulins. Since cleavage of the Fab fragment leads to a significant decrease in half-life, the latter provides information about the integrity of the immunoglobulin G (IgG) molecules. Therefore, a pharmacokinetic study of a novel preparation is required to determine safety and disposition in the target population. METHODS: Twenty-seven patients with chronic lymphocytic leukaemia (CLL) and 12 with multiple myeloma received intravenous infusions of IVIG containing antibodies against hepatitis B virus (anti-HBs; n= 15; 8960 IU), cytomegalovirus (anti-CMV; n = 9; 14,250 U) or varizella-zoster-virus (anti-VZV; n = 15; 6000 IU), respectively. Serum concentrations of viral antibodies were determined for 71 days during and after infusion. RESULTS: Maximum serum concentrations of anti-HBs, anti-CMV and anti VZV were observed at about 4 h (median) after start of the infusion. Total body clearances came to 0.14 +/- 0.08 ml/min (anti-HBs), 0.10 +/- 0.02 ml/ min (anti CMV) and 0.14 +/- 0.07 ml/min (anti-VZV). The terminal elimination half-lives were determined to be 25.34 +/- 8.34 days (anti-HBs), 24.66 7.28 days (anti-CMV) and 31.79 +/- 12.47 days (anti-VZV). Clinical chemistry parameters including C3- and C4-complement serum concentrations revealed no pathological changes, seroconversion for hepatitis B and C and HIV did not occur. CONCLUSIONS: The pharmacokinetic parameters of the IgG antibodies calculated after administration of the novel IVIG preparations to patients with CLL and multiple myeloma are in close agreement with data obtained from healthy volunteers and with values of native IgG, suggesting that the production process did not impair clinically relevant characteristics of the viral antibodies. PMID- 11497340 TI - Population pharmacokinetics of free carbamazepine in adult Omani epileptic patients. AB - OBJECTIVE: The aim of this study was to define the pharmacokinetic profile of free carbamazepine (F-CBZ) in adult Omani epileptic patients in order to improve on dosing schedules through population pharmacokinetic analysis using the NONMEM program. METHOD: Steady-state trough F-CBZ serum concentrations, carbamazepine (CBZ) dosing history and associated information were collected prospectively. RESULTS: Forty-eight patients with two or more available F-CBZ serum concentrations (total of 149 dose/serum concentration pairs) met our inclusion criteria. Patients were taking CBZ (200-1200 mg/day) in monotherapy. The analysis assumed a one-compartmental open model with first-order absorption and elimination. The apparent clearance (CL/F) and apparent volume of distribution (V/F) and their interindividual variabilities were estimated using the program. The population estimates for clearance (CL; modelled independently of dose) and volume of distribution were 13.2 +/- 0.6 l/h and 525 +/- 44 1, respectively. However, CL increased as a function of dosing rate and consequently was modelled as a linear function of steady-state concentration. In order to validate these results, the predictions of the population model were tested against data from 13 further patients subjected to the same inclusion criteria but who were not included in the original analysis. The predictions were good, being unbiased (P=0.31), and had an average deviation from the observed values of 18%. CONCLUSION: In order to establish steady-state dosage regimens, a population pharmacokinetic model is proposed, based on the patient's dose, to estimate the individual CL for an Omani epileptic patient receiving CBZ in monotherapy. PMID- 11497341 TI - Omapatrilat in patients with hepatic cirrhosis. Pharmacodynamics and pharmacokinetics. AB - OBJECTIVE: The pharmacodynamics and pharmacokinetics of omapatrilat, a member of a new class of cardiovascular compounds, the vasopeptidase inhibitors, were evaluated in subjects with hepatic cirrhosis (n = 10) and in healthy subjects (n = 10) matched for age, weight, gender and smoking history. METHODS: All subjects received omapatrilat 25 mg orally once daily for 14 days. Plasma renin and urinary atrial natriuretic peptide (ANP) levels were measured to assess the effect of omapatrilat on cirrhotic subjects. The effect of omapatrilat on blood pressure as well as changes in ANP and plasma renin levels were not altered by hepatic impairment. Pharmacokinetic parameters were determined from plasma omapatrilat concentrations. RESULTS: There were no significant differences between the two subject groups with regard to log-transformed area under the curve or maximum observed plasma concentration. Systemic accumulation was similar in the two groups. CONCLUSION: These results suggest, based on findings in otherwise healthy cirrhotic subjects, that no adjustment of standard dosing regimens is indicated for hypertensive patients with mild to moderate cirrhosis. PMID- 11497342 TI - Drug use during early pregnancy. The impact of maternal illness, outcome of prior pregnancies and socio-demographic factors. AB - OBJECTIVE: Socio-demographic factors have been associated with drug use in pregnancy. However, information on maternal illness has rarely been included in previous studies. The aim of this study was therefore to estimate the impact of maternal illness and prior pregnancy outcome on the use of analgesics/antipyretics, anti-infectives and antihistamines in early pregnancy controlling for socio-demographic factors. METHODS: One thousand nine-hundred and forty-five parous pregnant women from Norway and Sweden were included in an interview study. Information on drug use, obstetric history, maternal illness and socio-demographic factors was collected by specially trained midwives at gestational week 17 and form the basis for the present analyses. RESULTS: After control for socio-demographic factors, reported illness was associated with the use of analgesics/antipyretics, anti-infectives and antihistamines during early pregnancy. The use of analgesic/antipyretic and anti-infective drugs differed between study sites. Sociodemographic factors such as age, parity, marital status, education, occupation and smoking habits did not influence drug use in the multivariate analysis. CONCLUSION: Maternal illness during pregnancy was associated with drug use in early pregnancy. When studying factors related to drug use during pregnancy, it is important to include information on maternal illness. Socio-demographic factors may be confounders and lead to erroneous conclusions about the factors related to drug use. PMID- 11497345 TI - Antisense effects in the cell nucleus: modification of splicing. AB - The surprisingly small number of human genes, which has recently been estimated to be approximately 30,000, suggests that RNA processing, and in particular alternative RNA splicing, is in large part responsible for the diversity of gene products in human and mammalian cells. The ability to manipulate alternative splicing using antisense oligonucleotides, as demonstrated in several studies during the past year, makes it an important and attractive approach to altering gene expression. A review of these studies leads to the conclusion that antisense oligonucleotides, whether designed to affect the cytoplasmic mRNA or nuclear pre mRNA, function predominantly in the nucleus and not the cytoplasm. PMID- 11497344 TI - The zebrafish as a novel system for functional genomics and therapeutic development applications. AB - Unconventional antisense technology has entered the mainstream for both therapeutic and functional genomics applications in a variety of biological settings. Further development of this approach has been hampered by the high cost and limited information obtained with standard bioassays and animal models. The embryo of the zebrafish Danio rerio offers both biologists and technologists a new strategy that rapidly garners efficacy, toxicity and specificity data in an in vivo setting. This system has been used to optimize current antisense targeting methods, and it provides an ideal initial assay system for the development of new chemistries or other new gene targeting approaches. PMID- 11497346 TI - Phosphorodiamidate morpholino oligomers: favorable properties for sequence specific gene inactivation. AB - The phosphorodiamidate morpholino oligomers (PMOs) are a distinct class of oligonucleotide analogs. They bind to RNA and efficiently interfere with gene expression in a sequence-specific manner. Their non-ionic character combined with resistance to degradation in the body offer efficacy with minimal toxicity. Hence, the PMOs are emerging as an excellent approach to therapeutics that can be rationally designed based on target gene sequence data. PMID- 11497347 TI - Locked nucleic acids: a promising molecular family for gene-function analysis and antisense drug development. AB - Locked nucleic acids (LNAs) are a family of conformationally locked oligonucleotide analogs inducing unprecedented binding affinity towards DNA/RNA target sequences. Importantly, by virtue of the structural resemblance of LNAs to natural nucleic acid monomers, a combination of LNA chemistry with other oligonucleotide chemistries can be exploited to fine-tune the properties towards optimized antisense drug development and target validation technology. The first promising antisense results from experiments with LNA in living animals are described. PMID- 11497348 TI - Topical and transdermal delivery of antisense oligonucleotides. AB - Antisense oligonucleotides have great potential as therapeutic agents. As a result, a variety of chemistries have been developed to improve the efficacy of these molecules without compromising their specificity. Because the skin is such a large organ with extensive accessibility, it is a natural target for drug delivery. It is, however, an effective barrier, so physical and chemical methods to improve drug penetration have been developed. These enhancement techniques can be combined with modified oligonucleotide chemistries to provide sufficient levels of antisense activity either within the skin or systemically. This review will describe in vitro and in vivo experiments that demonstrate the potential of antisense oligonucleotides to treat both dermal and systemic disorders. PMID- 11497349 TI - Redirection of drug metabolism using antisense technology. AB - The cytochrome P450 (CYP) family is the most catalytically versatile component of the phase I oxidation metabolic pathway and participates in the metabolism of a large majority of drugs used in clinical practice. The inhibition of specific enzymes of this family can significantly alter the disposition and toxicity of substrate drugs by reducing and/or redirecting their metabolism. This review discusses the approaches available for CYP inhibition, with particular emphasis on the potential use of antisense phosphorodiamidate morpholino oligonucleotide strategies to inhibit human CYP3A4. Inter-individual variations of 10- to 50-fold have been reported in the activity of CYP3A4 enzyme, which contributes to the metabolism of more than half of all clinically relevant drugs. The application of antisense technology for inhibition of specific CYP enzymes can provide significant therapeutic benefits, including: (i) reduction of first-pass drug metabolism; (ii) reduction in drug dosage; (iii) selective reduction of toxic metabolites; and (iv) increased oral/topical drug bioavailability. The use of antisense morpholino oligonucleotide strategies to target CYP enzymes may result in safer and more uniform therapeutic applications. PMID- 11497351 TI - Perspectives on antisense therapy for the prevention of restenosis. AB - One of the potential clinical applications of antisense therapy is the prevention or treatment of restenosis following coronary interventions. Inhibition of several cellular proto-oncogenes have been shown to inhibit smooth muscle cell proliferation in vitro and to reduce neointimal thickening in vivo. The clinical applicability of antisense technology, however, remains limited due to a relative lack of specificity, slow uptake across the cell membrane and rapid intracellular degradation of the oligonucleotide. The one study in humans with c-myc antisense yielded a negative result with respect to restenosis after stent implantation. Recent studies have introduced phosphorothioate morpholino oligomers (PMO), which represent an unusual DNA chemistry with a six-membered morpholino ring instead of a deoxyribose sugar. In addition, the charged phosphodiester internucleotide linkage is replaced by an uncharged phosphorothioate. The PMOs are resistant to serum nucleases found in serum and exhibit a high degree of specificity and efficacy in both in vitro and cell-free translation studies. In vivo studies in four different animal models of restenosis demonstrated significant reduction of myointimal response. The combination of enhanced efficacy and greater specificity introduced by the PMO chemistry led us to re-examine the potential efficacy of a neutrally charged c-myc antisense approach for the prevention of restenosis. Clinical studies are underway to investigate safety and efficacy of local delivery of this latest generation of antisense to reduce restenosis after coronary stenting. PMID- 11497350 TI - Antisense oligonucleotides strategy in the treatment of hypertension. AB - Many kinds of drugs are used for the treatment of hypertension, but they are all short-acting (< or = 24 h) and patient compliance is poor. Gene therapy offers the advantage of producing long-term effects with high specificity, which should increase efficacy and reduce side effects. An antisense oligodeoxynucleotide (AS ODN) is a single-stranded oligonucleotide containing a gene-specific sequence of nucleotides, which is used to inhibit translation of mRNA. The application of AS ODNs for the treatment of hypertension began with targeting the renin-angiotensin system. Other genes, such as that coding for the beta1-adrenoceptor, have recently been targeted with AS ODNs in an attempt to reduce blood pressure. Strategies for the application of antisense technologies can be classified in two ways: the direct application of AS ODNs, and the production of AS by AS-cDNA inserted into viral vectors. Promising preclinical results from basic research have made feasible the possibility for antisense gene therapy of hypertension in the future. PMID- 11497352 TI - Hepatitis C IRES: translating translation into a therapeutic target. AB - Several approaches have been undertaken in the attempt to inhibit hepatitis C virus (HCV) translation. Antisense oligonucleotides (AS ONs) have proven to be invaluable in the characterization of the HCV internal ribosome entry site (IRES). Chemical modification of oligonucleotides has resulted in optimized stability and specificity. Artificial ribozymes have also been developed to target the HCV IRES. Both techniques have demonstrated efficacy in vitro and in vivo. Various studies have identified cellular cofactor proteins that are required for IRES function, which may present themselves as intervention targets. Recent experiments have revealed that the HCV IRES uses a novel mechanism of recruiting translational components. These new advances in understanding the mechanism of HCV translation could lead to the development of novel IRES inhibitor strategies. PMID- 11497353 TI - Technology evaluation: fomivirsen, Isis Pharmaceuticals Inc/CIBA vision. AB - Fomivirsen (ISIS-2922, Vitravene) is an antisense 21 mer phosphorothioate oligonucleotide with sequence complementarity to the coding region of the major immediate-early gene of human cytomegalovirus (CMV). Developed by Isis Pharmaceuticals Inc, fomivirsen is the first antisense oligonucleotide to receive approval for licensing and is marketed by Novartis' CIBA Vision. Fomivirsen is administered by intravitreal injection to AIDS patients for the treatment of CMV induced retinitis. In August 1998, the FDA approved the marketing of Vitravene for the local treatment of CMV retinitis [296420], [296780], and both European and Brazilian registration approval followed in the summer of 1999 [335238]. PMID- 11497354 TI - Technology evaluation: SGN-15, Seattle Genetics Inc. AB - Seattle Genetics is developing SGN-15 (BMS-182248, BR96-DOX), a monoclonal antibody-doxorubicin conjugate licensed from Bristol-Myers Squibb (BMS) for the potential treatment of cancer, which is currently in phase II trials [283767], [305881], [391927]. BMS had licensed technology from Enzon to help antibodies such as SGN-15 penetrate tumors more efficiently [174538]. BMS also had an R&D agreement with Genzyme for the production of SGN-15 in transgenic goats milk [177921]. In 1993, Ixsys (now Applied Molecular Evolution Inc) and BMS entered into a three-year collaboration to apply Ixsys's molecular design technology to BMS's monoclonal antibody research [190662]. Seattle Genetics licensed SGN-15 from BMS in April 1999 [283767]. PMID- 11497355 TI - To the question of originality of treatment of congenital hip dysplasias by active movement in the stirrups. 1959. PMID- 11497356 TI - Characteristic features of the acetabular labrum in healthy children. AB - Radial magnetic resonance images of the acetabular labrum were obtained on 40 hips of healthy children. There were no right-left or male-female differences. In children aged 11 years or younger, the labrum on the antero-superior weightbearing portion was triangular in shape, and there was an insular-shaped or linear high-intensity area inside; on the mid-superior portion, the labrum appeared as a regular triangular, low-intensity area; and on the postero-superior portion, it was flat. In children aged 12 and 13 years, the shape of the labrum in each portion was similar to that of the younger children, but the high signal intensity area on the antero-superior portion appeared less frequently. The size of the labrum relative to the femoral head was greater in younger children. PMID- 11497357 TI - Good results after treatment with the Frejka pillow for hip dysplasia in newborns: a 3-year to 6-year follow-up study. AB - Because there is no consensus with regard to the efficiency of the Frejka pillow in the treatment of hip joint dysplasia in newborns, the aim of the present study was to evaluate our results with this device. During the 3-year period 1988 to 1990, the Frejka pillow was used in 108 newborns with clinically unstable hips verified by ultrasonography. There were three treatment failures (2.8%), defined as infants who needed additional treatment with an abduction splint or hip-spica cast. Avascular necrosis of the femoral head occurred in one patient (0.9%). At an age of 3 years to 6 years, 85 of the children attended a follow-up examination. An intoeing gait was observed in 17% and slightly reduced hip mobility in 20% of the patients. Compared with normal children, the patients had somewhat lower coverage of the femoral head by radiography, indicated by a lower centre-edge angle and a higher migration percentage, but the coverage was within the normal range in all cases. The mean anteversion angle was larger than that of normal children but only three patients had abnormally high anteversion angles. In conclusion, the results with the Frejka pillow were good, with few treatment failures and complications, and it is the most simple abduction device for the parents to handle. More rigid devices like the von Rosen splint seem to involve a slightly lower failure rate, but a higher risk of avascular necrosis. Therefore, we recommend the Frejka pillow when treatment is started within a few days of birth. PMID- 11497358 TI - The flexion-adduction test: an early sign of hip disease. AB - This report introduces the flexion-adduction test as an additional diagnostic technique for hip pathology in children and young adults. Losses of flexion adduction, internal rotation, abduction and total flexion arc are all individually assessed in patients with confirmed hip pathology. The findings suggest that the "flexion-adduction" test is a simple, but sensitive, clinical procedure to detect the early signs of hip disease in older children and young adults. Although abduction and internal rotation are reportedly the most commonly restricted movements in frequent hip pathologies such as irritable hip, Legg Calve-Perthes disease and slipped epiphysis, we believe that the loss of flexion adduction is often the first affected range of movement. A statistical analysis of 87 cases from 1994 to 1997 is presented wherein the clinical loss of flexion adduction provided the initial suggestion of underlying hip pathology, and concludes that the loss of internal rotation, abduction and flexion arc are not statistically reliable tests for clinically diagnosing pathology individually. Often, despite a positive flexion-adduction test, the other ranges of movements remained unaffected or minimally affected in children and young adults with varying hip diseases, later confirmed through subsequent physical examinations or investigations. A thorough explanation of the flexion-adduction test is provided. PMID- 11497359 TI - Treatment of developmental dysplasia of the hip with the Pavlik harness: factors for predicting unsuccessful reduction. AB - To evaluate the risk factors for unsuccessful reduction, we reviewed 100 hips with developmental dysplasia of the hip treated with the Pavlik harness. We divided them into a successful reduction group (reduced without avascular necrosis) and an unsuccessful reduction group (reduced followed by avascular necrosis or unreduced). We compared Yamamuro's distance A, the acetabular angle and age at initial application of the Pavlik harness between the two groups. Our data suggest that a Yamamuro's distance A of 7 mm or smaller, an acetabular angle of 36 degrees or greater, and an age at initial application of 4 months or older are risk factors. PMID- 11497360 TI - Risk-benefit analysis of prophylactic pinning in slipped capital femoral epiphysis. AB - There is a broad and controversial discussion about the surgical procedure and the type of hardware for internal transfixation of the epiphysis and metaphysis in slipped capital femoral epiphysis (SCFE). Prophylactic pinning is even more controversial. One hundred and nine patients showing SCFE underwent a one-stage bilateral fixation of the epiphysis with three or four Kirschner wires (pins). From these 109 patients (69 male and 40 female), 94 had an unilateral slip and were operated prophylactically on the contralateral side. There were no complications such as avascular necrosis of the femoral head, chondrolysis, bone fracture, failure of metal implant, osteomyelitis or deep wound infection either at the time of surgery or at the minimum follow-up of 1 year with prophylactic pinning in SCFE. Therefore, we consider pinning allows for efficient stabilization, reliably preventing any progression of SCFE on the affected side and, furthermore, prevents the incidence of a secondary slip on the primarily nondisplaced contralateral side. The transfixation of epiphysis and metaphysis with Kirschner wires (pins) shows good subjective and objective long-term results compared with other surgical methods and implants. There is only a low morbidity rate with this method, because reoperations may only become necessary in the younger age group owing to normal growth of the femoral neck, compared with a high benefit from prophylactic surgical treatment of the nonaffected opposite side at the time of unilateral onset of the disease. The pins may no longer catch the epiphysis but further growth will allow for remodeling of the femoral head and for an optimal neck/shaft ratio. In case of further growth and relative shortening of the pins, refixation may become necessary. Therefore, we like to recommend the Kirschner-wire transfixation (pinning) of the epiphysis and metaphysis in patients with SCFE for primary treatment of SCFE as well as for prophylactic pinning of the contralateral side in one sitting. PMID- 11497361 TI - Evolution of scoliosis in six children treated with growth hormone. AB - During a period of 6 years to 11 years, the authors have been following six girls with scoliosis and treated with growth hormone (GH) for a growth insufficiency. The treatment with GH started after the discovery of the scoliosis for five patients. Three curve progressions have been observed, but always in the puberty period. Only one progression was noticed at the beginning of the GH treatment, but it was relieved with bracing. The results of this study do not permit one to conclude that a relation exists between GH treatment and scoliotic progression. This treatment is nevertheless not devoid of side effects, and a rigorous supervision is necessary. PMID- 11497362 TI - Posterior arch bifocal fracture of the atlas vertebra: a variant of Jefferson fracture. AB - Fracture of the atlas vertebra is rare in children. We report two paediatric cases of bifocal pedicular fracture of the posterior arch of C1. Evaluation was performed by nonenhanced computed tomography scan, which successively confirmed both diagnosis and healing. In both cases, nonoperative management was successful. PMID- 11497363 TI - Arthrotomy and open reduction of the displaced fracture of the femoral neck in children. AB - The objective of the study was to examine the efficacy of limited arthrotomy coupled with open reduction and internal fixation to minimize potential complications of vascular disruption of the capital femoral physis and femoral epiphysis. It was a patient outcome study. Skeletally immature patients with femoral neck fractures were treated according to a protocol in which all patients underwent limited arthrotomy with decompression of the intercapsular hematoma, open reduction and internal fixation. Radiographic and magnetic resonance imaging studies were carried out on patients to assess whether avascular necrosis developed in the postoperative period. Patients were followed for a mean length of 2 years 8 months, with a range of 2 years to 5.5 years. Twelve patients had good results and one had fair results using Ratliff criteria. No patient developed avascular necrosis (ischemic necrosis), coxa vara, nonunion or premature epiphyseal closure. This study supports the hypothesis that limited decompression of the intracapsular hematoma in children with femoral neck fractures may be a factor in obviating the subsequent appearance of ischemic disruption of the epiphysis and physis. PMID- 11497364 TI - Percutaneous pin fixation of intercondylar fractures in young children. AB - T-condylar fractures of the distal humerus are very rare in children. When they do occur they tend to affect those approaching skeletal maturity. Most agree that the undisplaced fracture can be managed conservatively. With increasing displacement and comminution the opinions tend to differ. As an alternative to open reduction and internal fixation we report the use of closed reduction and percutaneous pin fixation for displaced and comminuted T-condylar fractures in a series of three young children. In addition to the classical medial and lateral column fixation we report the use of a transcondylar wire to stabilise the distal fragments and discuss the use of intra-operative arthrography in low supracondylar fractures to exclude a missed intercondylar extension on plain xrays. Although small this series represents the largest series of children less than eight years of age. PMID- 11497365 TI - Transphyseal osseous bridges in experimental osteonecrosis of the femoral head of the rat. Histologic study of the bony bridges connecting the epiphyseal with the metaphyseal bony trabeculae through gaps in the physeal cartilage. AB - In view of the lifelong persistence of the physis, the femoral head of rats may serve to model Perthes disease and slipped capital femoral epiphysis. To produce osteonecrosis, the blood supply of one femoral head of 133, 6-month-old animals was severed by circumferentially incising the periosteum of the neck and cutting the ligamentum teres. The rats were killed 7 days to 90 days postoperatively. Associated with resorption of the necrotic bone and marrow, remodeling of the epiphysis was characterized by an ingrowth of vascularized fibrous tissue, formation of new bone and some cartilage, architectural deformation and flattening of the head. In 22 of 83 rats killed 30 days or more postoperatively, gaps in the continuity of the physeal cartilage were occupied by osseous bridges, connecting newly formed epiphyseal bony trabeculae with either the preexisting or newly formed metaphyseal osseous trabeculae. This healing mode may follow ischemic death of physeal chondrocytes or be owing to another mechanism, e.g., release of mediatory substances of inflammation. These findings raise the possibility that fixation of the healing epiphysis of a child's previously necrotic femoral head to the metaphysis occurs by transphyseal osseous growth in cases in which the physis is involved in the necrotic process. PMID- 11497366 TI - Hip subluxation and dislocation in Duchenne muscular dystrophy. AB - Although hip subluxation and dislocation occur commonly in neuromuscular diseases such as cerebral palsy, spinal muscular atrophy and myelomeningocele, they are not known to commonly affect patients with Duchenne Muscular Dystrophy. In this study, 15 out of 54 patients whose hip radiographs were reviewed showed unilateral subluxation, one had bilateral subluxation and three had unilateral dislocation. Having established that hip subluxation and dislocation develop frequently in patients with Duchenne Muscular Dystrophy, serial hip radiographs should be taken as has been advocated for cerebral palsy. The possible relationship between subluxation and pelvic tilt also calls for better control of sitting posture to prevent pelvic tilt. Spinal stabilization should be carried out at an early age when any scoliosis and pelvic tilt are still mild, so that progressive subluxation of the hip may be delayed or prevented, in addition to maintaining sitting balance and comfort. PMID- 11497367 TI - Propulsive function during gait in diplegic children: evaluation after surgery for gait improvement. AB - Seventeen patients with diplegic cerebral palsy were assessed by clinical examination and three-dimensional gait analysis before and after surgery to improve gait. Selection of surgical procedures was according to a fixed set of selection criteria. The average postoperative follow-up was 3.8 years (range, 2.6 5.7 years). Clinical examination revealed an improved range of motion for the ankle and no reduction in the power grade at the hip, knee and ankle after surgery. Kinematic parameters showed improved knee extension in stance and significant changes towards a normal ankle motion pattern postoperatively. Kinetic evaluation demonstrated that most of the total power during walking was generated at the hip, with the ankle contributing a small part. After surgery, patients walked faster with an increased power generation at the hip during first double support and at the ankle during push off. Power generation at the hip in stance is pointed out to be an important mechanism for propulsion during walking. PMID- 11497368 TI - Fibula pseudarthrosis revisited treatment with Ilizarov apparatus: case report and review of the literature. AB - A case of isolated congenital fibula pseudarthrosis with progressive valgus ankle deformity is reported. Valgus deformity can be corrected early by Ilizarov apparatus. Bone grafting is an essential part of treatment for union of the pseudarthrosis site. Internal splinting may be helpful in preventing further valgus deformity by maintaining continuity of fibula. Age of presentation has an important bearing on the choice of treatment. PMID- 11497369 TI - Limb lengthening in short stature patients. AB - A series of 140 patients with short stature operated on for limb lengthening (80 had achondroplasia, 20 had hypochondroplasia, 20 had Turner syndrome, 10 had idiopathic short stature due to an undemonstrated cause, 5 regarded their stature as too short, and 5 had a psychopathic personality due to dysmorphophobia that had developed because of their short stature) was reviewed. All patients underwent symmetric lengthening of both femora and tibiae; 10 of these achondroplastic patients underwent lengthening of the humeri. We carried out the 580 lengthening procedures by means of three different surgical techniques: 440 callotasis, 120 chondrodiatasis and 20 mid-shaft osteotomy. In the 130 patients with a disproportionate short stature, the average gain in length was 18.2 +/- 3.93 cm: 43.8% had complications and 3.8% had sequelae; the average treatment time was 31 months. In the 10 patients with proportionate short stature, the average gain in length was 10.8 +/- 1.00 cm: 4 experienced complications and none had sequelae; the average treatment time was 21 months. Patients who underwent lengthening of the upper limbs experienced an average gain in length of 10.2 +/- 1.25 cm: the average treatment time was 9 months and none of them experienced any complications or sequelae. The authors discuss how difficult it is to achieve the benefits of this surgery: they underline the strong commitment on the part of the patients and their families, the time in the hospital, the number of operations and, above all, the severity of those permanent sequelae that occurred. PMID- 11497370 TI - Fracture stiffness in callotasis determined by dual-energy X-ray absorptiometry scanning. AB - Assessing healing after distraction limb lengthening is essential to manage patients undergoing callotasis for leg lengthening or bone transport. Direct measurement of fracture stiffness can assess healing but the equipment may not be available. In addition, it requires removal of the fixator, which may be complicated for ring fixators. The present study investigates whether an equivalent measure of healing can be based on the mineral density pattern from dual-energy X-ray absorptiometry (DXA) scans. Nine consecutive patients undergoing callotasis were studied. Bending stiffness of the distraction segment was measured and DXA scans were performed regularly starting 6 weeks after completing distraction. In all, 23 simultaneous readings of bending stiffness and DXA scans were obtained. All density patterns showed a distinct minimum value of bone mineral density. We found a high and significant correlation between fracture bending stiffness and the square of the total mineral content at the location of minimum bone density (r2 = 0.77, P < 0.001). We conclude that DXA scans can be used reliably and effectively to determine fracture bending stiffness, valuable for determining both time of frame removal and delay in union. PMID- 11497371 TI - Therapy in unilateral sirenomelia: report of one case. AB - A 17-month-old male infant was referred to us with a unilateral sirenoid malformation in 1970. Besides urogenital and lumbosacral vertebral defects, the right knee was flexed 160 degrees, and the hip externally rotated and abducted. The thigh and calf were connected by a soft tissue bridge with the heel fixed to the ischial region. Arteriography showed that the foot and lower leg were supplied by the profunda femoris artery through the soft tissue bridge. The lower leg had only a very small supply from the popliteal artery. Therefore, an amputation by disarticulation at the knee joint was performed. A few months later, in addition, the hip flexion contracture was released. The boy was taught to walk with a prosthesis. At the age of 31 years, he has a sedentary job and is more troubled by his urogenital than his orthopaedic defects. He prefers to walk with crutches and for a few years has no longer used his prosthesis. PMID- 11497372 TI - Clinical evaluation of patients with idiopathic juvenile osteoporosis. AB - The aim of this study is to evaluate the occurrence of idiopathic juvenile osteoporosis by clinical, radiographic and biochemical methods in 7 patients aged 10 years to 18 years. The prevailing clinical symptoms included pain in the lower limbs and spine, gait disturbances and weak muscles; confirmed fractures were seen in four out of seven patients. In all patients, bone mineral density was determined by dual-energy X-ray absorptiometry and calcium-phosphorus metabolism assays were performed. Clinical evaluation of our patients revealed a wide variety of classic clinical symptoms, and also radiographic and biochemical features. In this complex evaluation, we can distinguish two types of disease occurrence, mild and acute. PMID- 11497373 TI - Cysteine proteases of Porphyromonas gingivalis. AB - The cysteine proteases of Porphyromonas gingivalis are extracellular products of an important etiological agent in periodontal diseases. Many of the in vitro actions of these enzymes are consistent with the observed deregulated inflammatory and immune features of the disease. They are significant targets of the immune responses of affected individuals and are viewed by some as potential molecular targets for therapeutic approaches to these diseases. Furthermore, they appear to represent a complex group of genes and protein products whose transcriptional and translational control and maturation pathways may have a broader relevance to virulence determinants of other persistent bacterial pathogens of human mucosal surfaces. As a result, the genetics, chemistry, and virulence-related properties of the cysteine proteases of P. gingivalis have been the focus of much research effort over the last ten years. In this review, we describe some of the progress in their molecular characterization and how their putative biological roles, in relation to the in vivo growth and survival strategies of P. gingivalis, may also contribute to the periodontal disease process. PMID- 11497374 TI - Genetic competence and transformation in oral streptococci. AB - The oral streptococci are normally non-pathogenic residents of the human microflora. There is substantial evidence that these bacteria can, however, act as "genetic reservoirs" and transfer genetic information to transient bacteria as they make their way through the mouth, the principal entry point for a wide variety of bacteria. Examples that are of particular concern include the transfer of antibiotic resistance from oral streptococci to Streptococcus pneumoniae. The mechanisms that are used by oral streptococci to exchange genetic information are not well-understood, although several species are known to enter a physiological state of genetic competence. This state permits them to become capable of natural genetic transformation, facilitating the acquisition of foreign DNA from the external environment. The oral streptococci share many similarities with two closely related Gram-positive bacteria, S. pneumoniae and Bacillus subtilis. In these bacteria, the mechanisms of quorum-sensing, the development of competence, and DNA uptake and integration are well-characterized. Using this knowledge and the data available in genome databases allowed us to identify putative genes involved in these processes in the oral organism Streptococcus mutans. Models of competence development and genetic transformation in the oral streptococci and strategies to confirm these models are discussed. Future studies of competence in oral biofilms, the natural environment of oral streptococci, will be discussed. PMID- 11497375 TI - Pathogenetic factors in Sjogren's syndrome: recent developments. AB - The study of pathogenetic factors in Sjogren's syndrome [SS] has been problematic, given the overall paucity of coherent data that integrate basic research with clinical findings. The presumed autoimmune nature of SS suggests T cells, autoantibodies, and cytokines as possible immune factors in the initiation and progression of SS. Recent work on programmed cell death (apoptosis) in SS and its models suggests this as a fourth potential mechanism of disease. These four areas of SS research are reviewed with an emphasis on the most recent findings related to mechanisms of disease. New findings confirm the potential for antigen presentation to T-cells in the salivary glands, as well as involvement of other adhesion molecules with respect to T-cell functions. Restrictions on the receptor repertoires of infiltrating T-cells are discussed, as are new findings on antigenic specificities of these cells. New findings on the specificities of autoantibodies observed in SS are reviewed with an eye toward potential mechanisms for depression of exocrine secretory capacity. Stimulating new findings concerning cytokine production in salivary and lacrimal gland are noted. Particular points of interest with regard to apoptosis include the wide range of values obtained for apoptotic activity in SS and its models, and potential means of resolving discongruent results and the study of factors influencing apoptosis are discussed. PMID- 11497376 TI - The impact of microgravity on bone metabolism in vitro and in vivo. AB - Exposure to microgravity has been associated with several physiological changes in astronauts and cosmonauts, including an osteoporosis-like loss of bone mass. In-flight measures used to counteract this, including intensive daily exercise regimens, have been only partially successful in reducing the bone loss and in the process have consumed valuable work time. If this bone loss is to be minimized or, preferably, prevented, more effective treatment strategies are required. This, however, requires a greater understanding of the mechanisms through which bone metabolism is affected by microgravity. Various research strategies have been used to examine this problem, including in vitro studies using bone cells and in vivo studies on humans and rats. These have been conducted both in flight and on the ground, by strategies that produce weightlessness to mimic the effects of microgravity. Overall, the majority of the studies have found that marked decreases in gravitation loading result in the loss of bone mass. The processes of bone formation and bone resorption become uncoupled, with an initial transitory increase in resorption accompanied by a prolonged decrease in formation. Loss of bone mass is not uniform throughout the skeleton, but varies at different sites depending on the type of bone and on the mechanical load received. It appears that the skeletal response is a physiologic adaptation to the space environment which, after long space flights or repeated shorter ones, could eventually lead to significant reductions in the ability of the skeletal tissues to withstand the forces of gravity and increased susceptibility to fracture. PMID- 11497377 TI - Changing paradigms in implant dentistry. AB - This review focuses on five paradigms of implant dentistry which have undergone considerable modifications in recent years. An attempt was made to select and include all the relevant citations of the past 10 years. These five paradigms document the debate in the clinical and scientific community and include the aspects of (1) smooth vs. rough implant surfaces, (2) submerged vs. non-submerged implant installation techniques, (3) mixed tooth-implant vs. solely implant supported reconstructions, (4) morse-taper abutment fixation vs. butt-joint interfaces, and (5) titanium abutments vs. esthetic abutments in clinical situations where esthetics is of primary concern. PMID- 11497378 TI - Analysis of heavy metals in atmospheric particulates in relation to their bioaccumulation in explanted Pseudevernia furfuracea thalli. AB - Lichens explants and airborne particulates were analysed for Cr, Zn, Cd, Pb, Ni, Fe, Mn and Cu content in a rural site in the province of Vercelli (Piedmont, N. Italy). Thalli of the fruticose lichen Pseudevernia furfuracea were collected from an uncontaminated area and exposed for one year using two different techniques. Particulates were collected using a continuously operating particulate sampler set up near the lichens explanted throughout the experiment survey. The results confirm both the importance of assessing contamination in rural areas corroborated by the analysis of the presence of the heavy metals in particulates in relation to the main wind directions and the validity of this lichen as biomonitor of airborne metal pollution, although meaningful differences were pointed out between the two different techniques of exposure. The significant correlation between most of the metals accumulated by lichens and the metal air concentrations shows a good evidence of their ability to represent atmospheric contamination. PMID- 11497379 TI - Testing the validity of a critical sulfur and nitrogen load model in southern Ontario, Canada, using soil chemistry data from MARYP. AB - The validity of a steady-state mass balance model (Arp et al., 1996; referred to as ARP) was tested using physicochemical soil data from the Monitoring Acid Rain Youth Program (MARYP). Four ARP sites were matched with ten MARYP sites according to proximity, bedrock type and subsoil pH to test the validity of the ARP model for critical load exceedances. Soil solution pH, base concentration and A1 concentration from MARYP sites, which were well matched to ARP sites, validated the modelled critical load exceedances. Higher exceedance areas were associated with more acidic pH and lower base and higher A1 concentrations from matched MARYP sites and vice versa. One ARP site was inappropriately matched with MARYP sites and could not be validated using base and A1 concentrations. This study also confirmed the southern limit of the zero critical load exceedance isopleth from the model. However, variability of the other exceedance isopleths was noted due to the limited number of sites used in the model. The validation of these sites in the ARP model and the zero critical load exceedance isopleth nonetheless allows greater confidence in using this model as a management tool for acidic deposition. PMID- 11497380 TI - Mulberry-silkworm food chain--a templet to assess heavy metal mobility in terrestrial ecosystems. AB - Assessment of the food chain mobility of heavy metals in the natural ecosystem receives more attention nowadays. In the present study, mulberry-silkworm food chain has been focused as a templet to assess the biomobility of heavy metals in soil-higher plant-insect hierarchy. Both in the case of Cd and Cu treatments, higher mobility was observed in the level-1 (soil-root) followed by level-3 (leaf larva), level-4 (larvae-faecal) and level-2 (root-leaf). Consequently, roots accumulated more amounts of Cd and Cu, with a limited transport to the leaves. Among the two metals (Cd and Cu) tested, in the plant, the transfer potential of Cd exceeds that of Cu. Whereas in the case of leaf-larval transfer, Cu precedes over Cd. Accumulation of Cd and Cu in all the levels (1-4) tested showed a concentration dependent increase, except in the level 4 (larva-faecal) of Cd treatment where a declining trend was noticed. PMID- 11497381 TI - Multiple-point variance analysis for optimal adjustment of a monitoring network. AB - The selection of sampling sites is one of the major tasks in the design of a monitoring network. Many environmental networks suffer from either insufficient information or redundant information. This study presents a new, effective algorithm that addresses the issues of insufficient and reduction information. The new algorithm is denoted as Multiple-Point Variance Analysis (MPV). MPV includes both Multiple-Point Variance Reduction Analysis (MPVR) for adding information-effectives sites, and Multiple-Point Variance Increase Analysis (MPVI) for deleting information-redundant sites. The MPVR and MPVI equations are verified under two hypothetical cases. The optimal procedures of this new algorithm include determination of simultaneous additions or deletions of groups of sampling points. These proposed optimization procedures eliminate the need for any spatial discretizations or sequential selections. The efficiency of these optimal procedures is tested under actual field conditions. The results show that the optimal MPV is an effective tool for adjustment of existing sampling networks. PMID- 11497382 TI - Honeybees and their products as potential bioindicators of heavy metals contamination. AB - The concentrations of three representative heavy metals (cadmium, chromium and lead) were measured by atomic absorption spectroscopy in honeybees and in apiary's products (honey, pollen, propolis, and wax). Samples were collected from five different sampling points: four from areas surrounding the city of Rome, and the fifth in the city center which receives intense vehicular traffic. All apiaries employed for this study were specifically constructed without any metal part in order to avoid the risk of contamination of the assayed materials. Sample collection was conducted over a 3-month period (6 samplings for honey and pollen, 3 sampling for propolis and wax, 2 samplings for honeybees, all of which were collected in duplicate). Experimental data revealed, in general, statistically significant differences between the background levels of heavy metals recorded from the reference sites and the levels measured in the site located in the center of the city of Rome. These results indicate that honeybees and, to a lesser extent, some of their products (pollen, propolis, wax, but not honey), can be considered representative bioindicators of environmental pollution. PMID- 11497383 TI - Nitrogen isotope ratios in pine bark as an indicator of N emissions from anthropogenic sources. AB - The article describes the use of Scots pine bark to identify nitrogen sources in eastern Germany, as well as background areas in Russia and Bulgaria, by using natural isotope ratios of total nitrogen (Nt) and individual N compounds such as ammonium (NH4+), nitrate (NO3-) and amid nitrogen (amide-N). The samples collected were analysed using an elemental analyser in connection with a gas isotope mass spectrometer (EA-IRMS). Natural 15N abundances in pine bark from impact areas suggest that the ammonium accumulated on the surface of the bark is released from livestock management. Bark of Scots pines growing near agricultural land had highly depleted delta15Nt values (between -8 and -12 one thousand percent), while bark from background areas (unpolluted areas) displayed slightly negative delta15Nt values (mean 15Nt = -3.8 one thousand percent). It is assumed that part of the N adsorbed on the bark surface is mainly derived from ammonia (mean 15Nt = -40.3 one thousand percent) escaping from livestock housing and during the application of manure. This assumption is confirmed by experiments under controlled conditions in which manure samples were spread on soil. In addition, temporal and spatial variations of 15Nt abundances in pine bark from various locations in eastern Germany as well as pine stands in Nature Park Dubener Heath are discussed. PMID- 11497384 TI - What, why and how: time-consuming but inevitable questions in ethical decision making. PMID- 11497385 TI - The development of palliative care at the University of Texas M.D. Anderson Cancer Center. PMID- 11497386 TI - Ethical decision-making on communication in palliative cancer care: a personalist approach. AB - Perhaps one of the main ethical dilemmas physicians face in cancer medicine is the question of truthfulness with terminally ill cancer patients. Reluctance to share the truth with the patient about his or her diagnosis and/or prognosis is frequently associated with cultural pressures. Based on two cases, the authors illustrate how ethical analysis can help in solving dilemmas related to truth disclosure to terminally ill cancer patients and their families. A personalist approach reveals that the often-adduced conflict between nonmaleficence/beneficence and autonomy with regard to truth telling originates from a narrow understanding of the concept of autonomy. This confrontation is, therefore, more apparent than real. A brief review of the main ethical systems and the results of their application to clinical decision-making follow the discussion of the cases. PMID- 11497387 TI - Major depression and demoralization in cancer patients: diagnostic and treatment considerations. AB - Major depression and demoralization are very common in patients with cancer. A discussion of the diagnostic specificity of major depression and demoralization (also known as adjustment disorder) is presented here, followed by a review of some effects of comorbid depression and cancer. Finally, there are a brief review of studies of antidepressant pharmacotherapy in cancer patients, a treatment algorithm for antidepressant therapy, and suggestions for treatment of demoralization. PMID- 11497388 TI - Potential role of the NK1 receptor antagonists in chemotherapy-induced nausea and vomiting. AB - Better tolerated and more effective means of controlling chemotherapy-induced nausea and vomiting have been introduced over the past decade. Despite the progress made, incompletely controlled emesis is a persistent problem for significant numbers of patients receiving chemotherapy. Efforts to improve antiemetic control further are ongoing. The most interesting new class of antiemetics under development focuses on antagonism of the neurotransmitter substance P. Substance P exerts its effects by binding to the tachykinin neurokinin NK1 receptor. A number of selective antagonists of the NK1 receptor have been synthesized and, when used in preclinical models, have demonstrated an ability to antagonize the emetic effects of a number of stimuli, including chemotherapy agents such as cisplatin. Over the past 3 years, results of the initial studies evaluating this class of agents for cisplatin-induced emesis in cancer patients have begun to appear. These agents have been well tolerated. As single agents, they appear to be no more effective than 5-HT3 receptor antagonists in preventing acute cisplatin-induced emesis. Their real value may be found in combination with existing agents and in the treatment of delayed emesis. The results of ongoing clinical trials will hopefully define the utility and appropriate place for this new class of agents in the management of chemotherapy induced nausea and vomiting. PMID- 11497389 TI - Dimensions of fatigue during radiotherapy. An application of the Multidimensional Fatigue Inventory. AB - The aim of the present study was to describe fatigue in a group of irradiated cancer patients. Fatigue was assessed prospectively in 81 Swedish cancer patients, using the Multidimensional Fatigue Inventory (MFI-20), which was also validated in the study, the Karolinska Sleepiness Scale and a Borg CR10 scale before curative radiotherapy, at the end of therapy and 1 and 3 months after treatment. The mean age of these patients was 56 years; 90% were female; and 65% were being treated for cancer of the breast. The absorbed radiation dose was 50 Gy or lower in 90% of the patients. A clear peak in experienced fatigue according to the three scales was seen at the end of treatment. In the MFI-20 the dimensions of General fatigue, Physical fatigue and Reduced activity, were more pronounced than the aspects of Mental fatigue and Reduced motivation. Comparison with other studies using the same instrument in other irradiated patients revealed a similar pattern. The internal consistency of the Swedish version of the MFI-20 was good. The MFI-20 is shown to be a valid instrument for assessing fatigue in patients undergoing radiotherapy. PMID- 11497390 TI - Quality of life measurement in the palliative management of advanced cancer. AB - A 4-week quality of life (QOL) study was conducted in 157 consecutive advanced cancer patients to compare five QOL measures and to assess changes in QOL during the course of the study. The study patients were representative of the advanced cancer population in the USA, and were similar to the advanced cancer population with respect to age and gender. Global QOL measures appear to be as effective as multidimensional measures in assessing QOL in patients with advanced cancer. Generally, QOL was rated as satisfactory and did not change markedly during the study period. PMID- 11497391 TI - Manifest Anxiety Scale for evaluation of effects of granisetron in chemotherapy with CDDP and 5FU for head and neck cancer. AB - The level of anxiety was examined before treatment by means of the Manifest Anxiety Scale (MAS) in 41 patients with squamous cell carcinoma of the head and neck. They received 5 days of neoadjuvant chemotherapy with cisplatin (CDDP) and 5-fluorouracil (5FU). Granisetron (KYT) was administered daily from day 1 to day 5. Nausea, vomiting, appetite, and well-being were assessed during and after chemotherapy. The relation between the effects of KYT and anxiety was studied. Seventeen patients were proven to have anxiety and were compared with the other 24 patients. In patients with anxiety, the percentage well-being was significantly lower on days 1 and 2 (P=0.008, 0.001). The rate of freedom from nausea was significantly lower from day 4 to day 9 for anxiety patients (P=0.010 0.050). The percentage of anxiety patients without loss of appetite was significantly lower from day 6 to 9 (p=0.001-0.020). The rate of freedom from vomiting was significantly lower on days 4, 5 and 7 for anxiety patients (P=0.024, 0.024, 0.014). The results indicate that the effect of KYT was significantly lower from day 3 to day 7 for anxiety patients (P=0.008-0.045). The anxiety group had significantly poorer well-being at the beginning of chemotherapy, and were not responsive to KYT in the delayed phase. Our results prove that anxiety patients show delayed emesis, and the administration of KYT is considered insufficient. It may be important to co-administer a tranquilizer to any patient who exhibit anxiety as defined by the MAS, in order to reduce delayed emesis. PMID- 11497392 TI - Piperacillin, beta-lactam inhibitor plus gentamicin as empirical therapy of a sequential regimen in febrile neutropenia of pediatric cancer patients. AB - The beta-lactam/beta-lactamase inhibitor combinations are a good choice for empirical antimicrobial therapy in febrile neutropenic patients, because their antibacterial spectra include both gram-negative and gram-positive pathogens. This trial was initiated to assess the efficacy and safety of piperacillin with the beta-lactam inhibitors sulbactam (PSG group) or tazobactam (PTG group) and gentamicin as initial therapy in febrile neutropenia of pediatric patients. In a prospective study, 239 episodes of fever and neutropenia were analyzed for the clinical and microbiological response dependent on infection etiology and treatment group: 66.5% of episodes were classified as fever of unknown origin (FUO) and 33.5%, as microbiologically or clinically documented infections; 19.2% of all episodes were due to bacteremia, predominantly caused by gram-positive organisms (69.6%). The response to the initial therapy was 55.2% overall and 65.4% in episodes of FUO with a significant higher success rate in the PSG group than in the PTG group (70.1% vs. 52.4%, P=0.039), and 35.0% in documented infections. In episodes with documented infection longer duration of fever and antimicrobial therapy was recorded than for FUO episodes. Four patients died of causes related to infection. Fever relapse occurred in 26 episodes (11.1%), predominantly in patients who were still neutropenic. Toxic side effects were minimal. The initial therapy of piperacillin with sulbactam or tazobactam in combination with gentamicin is well tolerated, and its efficacy is comparable to that of other combination therapies or of monotherapy with beta-lactam antibiotics in pediatric neutropenic cancer patients. PMID- 11497393 TI - Outpatient treatment of multiple myeloma with a combination of vincristine, Adriamycin and dexamethasone. AB - Patients with relapsed multiple myeloma (MM) have been shown to respond to a combination therapy consisting of vincristine, Adriamycin (doxorubicin) and high dose dexamethasone (VAD). Because of the low hematological toxicity of the VAD regimen, this combination is frequently chosen for tumor reduction prior to high dose therapy and blood stem cell transplantation. This study was designed to examine the efficacy and complications of outpatient VAD treatment. Over a period of 6 years, 103 outpatients with MM were treated with VAD chemotherapy administered by microprocessor-controlled infusion pumps via intravenous polyurethane catheters equipped with a safety valve. Response to treatment, treatment-associated complications and infections were documented and analyzed. In 85 of the 103 patients, tumor reduction by more than 25% was found. In 8 patients an occlusion occurred as a result of kinking of the central venous catheter in the subcutaneous segment. In two treatment cycles the infusions had to be stopped because of irreversible catheter occlusion. Twenty patients were hospitalized because of complications, which were infectious in 12 and noninfectious in 8. Severe infectious complications (> or =WHO grade III) occurred in 5.6% of the treatment cycles. Thus, continuous infusion of VAD over 96 h can be performed on an outpatient basis with a low complication rate. PMID- 11497394 TI - The use of corticosteroids in home palliative care. AB - Evidence for the effectiveness of corticosteroids in palliative care is anecdotal, and more information is required. From January to December 1999 a total of 376 consecutive patients admitted to a home palliative care program were longitudinally surveyed. Patients who started a corticosteroid treatment after admission on the basis of common indications prescribed by their home care physicians were selected. Fifty patients were enrolled in the study. Dexamethasone, in doses ranging from 4 to 16 mg, was the drug of choice. Corticosteroids were found to be effective in anorexia, weakness, headache, and nausea and vomiting. The reduction of symptom intensity was achieved in less than 3 days on average. However, no great advantages were found in terms of controlling drowsiness or confusional states associated with advanced illness because of cerebral involvement. It can be concluded from this study that: (a) corticosteroids may be effective in controlling anorexia, weakness, headache, and nausea and vomiting associated with cerebral involvement or bowel obstruction; (b) they should be stopped if no therapeutic effect has become evident within 3-5 days; (c) the treatment is not useful when given in the presence of severe neurological impairment resulting from the advanced stage of disease; (d) the range of adverse effects was acceptable for limited periods and in the circumstances in which the preparations were used in this study; and (e) corticosteroids may have an adjuvant role in potentiation of analgesic drugs. These findings will be very useful in the planning of future controlled studies designed to yield evidence-based data on the role of corticosteroids in the relief of specific symptoms. PMID- 11497395 TI - Giving bad news: a GPs' narrative perspective. AB - The purpose of this study was to explore GPs' perspectives on giving bad news during consultations. To this end, 168 GPs were asked to recall, and record on the first page of a questionnaire, an occasion when they had given medically related bad news to a patient. The stories were analysed with a qualitative and interpretative approach. Two axes, each with a semantic polarity, were identified: a relational axis (semantic polarity: escape vs accompanying) and an ethical axis (semantic polarity: the doctor's choice vs the patient's choice). Furthermore, two main topics appeared to be common to almost all the narratives: the need to reassure the patient and the account of the doctor's emotions. Two different relational patterns appear to be described by doctors. A substantial number of GPs implicitly describe a disease- or doctor-centred consultation: in these cases the physicians refer to signs and symptoms, diagnosis and treatments; they decide for themselves whether to tell the truth or not. On the other hand, a smaller number describe consultations that could be defined as patient centred: these doctors consider that their duty of care for the individual ill person is paramount and try to respect the patient's right to decide. In both these relational patterns, GPs feel it is a fundamental professional duty to reassure the patient; furthermore, they feel the most difficult aspect is managing their own emotional responses. PMID- 11497396 TI - Clinical benefits of lenograstim in patients with neutropenia due to chemotherapy for multiple myeloma (MM). AB - The object of this study was to determine the efficacy and safety of glycosylated recombinant human granulocyte colony-stimulating factor (rHuG-CSF; lenograstim) after combination chemotherapy consisting of ranimustine, vindesine, melphalan and prednisolone (MCNU-VMP). One hundred thirty-nine consecutive patients with newly diagnosed multiple myeloma (MM) were allocated at random to a lenograstim group (n = 70) or a placebo group (n = 69). Patients were treated with two cycles of MCNU-VMP, and either lenograstim (2 microg/kg daily, s.c.) or placebo was administered from the day neutrophils decreased to less than 1.000/microl and was discontinued when neutrophils exceeded 5,000/microl. The median duration of neutropenia (neutrophils under 1,000/microl) was significantly shorter for the lenograstim group than the placebo group (2 days vs 9 days in the first cycle; 1 day vs 13 days in the second cycle). The incidence of febrile neutropenia in the first cycle was significantly lower in the lenograstim group than in the placebo group (9.2% vs 30.4%). No life-threatening infections were observed in either group. The two cycles of MCNU-VMP therapy were completed in 90.8% of the patients, and a higher average relative dose intensity (ARDI; 0.94) was achieved in the lenograstim group. The tumor response rate of the lenograstim group (57.8%) was higher than that of the placebo group (43.1%), but the difference was not statistically significant (chi2 = 2.634, df = 1, P = 0.105). Lenograstim was well tolerated, and no unexpected adverse events occurred. Lenograstim proved effective in controlling chemotherapy-induced neutropenia in MM patients under MCNU-VMP therapy. PMID- 11497397 TI - Controlled study of the efficacy of clavulanic acid-potentiated amoxycillin in the treatment of Chlamydia psittaci in cats. AB - Twenty-four specific pathogen-free cats were inoculated with 3 x 10(3) infectious units of a field isolate of Chlamydia psittaci on to the corneal surface. Seven days later they were assigned randomly to three groups of eight and treated orally for 19 days with either clavulanic acid-potentiated amoxycillin, doxycycline or a placebo. Both treated groups responded rapidly, with a marked reduction in isolation rates and clinical scores which were significantly lower than in the placebo group within two and four days, respectively. After two days the group treated with potentiated amoxycillin had a significantly lower isolation score than the group treated with doxycycline. Forty days after they were infected the clinical signs recurred in five of the eight cats treated with potentiated amoxycillin, but a four-week course of potentiated amoxycillin resulted in a complete clinical recovery with no evidence of a recurrence for six months. PMID- 11497398 TI - Intrauterine device for contraception in dogs. AB - A new intrauterine device for contraception was tested on nine bitches. After it had been implanted, the bitches were mated but none of them became pregnant. Over a two-year period no side effects were observed, except in a bulldog bitch in which signs of oestrus persisted until the device had been removed. PMID- 11497399 TI - Comparison of simultaneous changes in electroencephalographic and haemodynamic variables in sheep anaesthetised with halothane. AB - Haemodynamic responses are routinely used for monitoring anaesthesia, but they can be elicited at spinal or brainstem levels and therefore do not provide information on cerebrocortical activity. To test the possibility that electroencephalography (EEG) might provide an objective assessment of unconsciousness and depth of anaesthesia the haemodynamic and EEG data from 17 ewes anaesthetised with halothane were analysed. Data were recorded at least every five minutes. An increase of 10 per cent or more in heart rate, systolic arterial pressure (SAP), or mean arterial pressure (MAP) was defined as a positive haemodynamic response to surgical stimulation. Simultaneously recorded EEG variables included delta-, theta-, and beta-frequency band power, power ratios (theta/delta, alpha/delta, beta/delta), 80 per cent spectral edge frequency (SEF 80), and median frequency. In 11 of the ewes, a positive haemodynamic response was accompanied by a significant decrease in delta activity and significant increases in alpha/delta ratio, beta/delta ratio, and SEF 80, and there were significant correlations between the changes in heart rate, and delta activity and beta/delta ratio. In the other six ewes, there were no significant EEG responses in association with haemodynamic changes of less than 10 per cent. PMID- 11497400 TI - Chronic inflammatory bowel disease of growing sheep. PMID- 11497401 TI - Diagnosis of pericardial effusion in birds by ultrasound. PMID- 11497402 TI - Indirect ELISA for the diagnosis of brucellosis in water buffaloes (Bubalus bubalis) in Italy. PMID- 11497403 TI - Pathological investigations on anthracosis in cattle. PMID- 11497404 TI - Ruptured dissecting aortic aneurysms in two red lechwe (Kobus leche). PMID- 11497405 TI - Anthelmintic-resistant nematodes in sheep in Scotland. PMID- 11497406 TI - Idiopathic mouth ulcers in sheep, cattle and horses. PMID- 11497407 TI - Idiopathic mouth ulcers in sheep, cattle and horses. PMID- 11497408 TI - First report of Mycobacterium bovis in a muntjac deer. PMID- 11497409 TI - Psoroptes species in alpacas. PMID- 11497410 TI - Equine magnetic resonance imaging research. PMID- 11497411 TI - Rod-/L-cone and rod-/M-cone interactions in electroretinograms at different temporal frequencies. AB - We recorded electroretinograms (ERGs) under stimulus conditions that only modulated one cone type: either the L- or the M-cones. In these conditions the rods were also modulated. We measured the ERG responses at different temporal frequencies. A simple model that assumes that the first harmonic components of the responses are the result of a vector addition of rod- and cone-driven ERG responses can explain the data satisfactorily for temporal frequencies at and above 6 Hz. From fits of the model to the data, estimates of the gains and phases of the rod- and cone-driven responses can be obtained. At 6 Hz, the fundamental responses originate exclusively in the rods. The gains of the cone-driven responses are substantial at the other temporal frequencies, are maximal at 12 or 18 Hz, and then decrease with increasing temporal frequencies. The gains of the rod-driven responses decrease more steeply with increasing temporal frequencies than the cone gains. Furthermore, the rod and cone phases decrease approximately linearly with temporal frequency, suggesting that they are mainly determined by a response delay. The response delay in the rod-driven ERGs is larger than in the cone-driven ERGs. PMID- 11497412 TI - Intraretinal analysis of the a-wave of the electroretinogram (ERG) in dark adapted intact cat retina. AB - It has often been assumed that the recovery of the a-wave from its trough is caused by the intrusion of the b-wave. This study examined the recovery following the a-wave trough using intraretinal recordings in dark-adapted intact cat retina. Adult cats were anesthetized and paralyzed. The vitreal ERG was recorded between the vitreous humor and a reference electrode near the eye. Intraretinal recordings were made by referencing a microelectrode to the vitreal electrode. Bright flashes of diffuse white light were used to elicit a- and b-waves. Intravitreal injections of 2-amino-4-phosphonobutyrate (APB), cis 2,3-piperidine dicarboxylic acid (PDA), and kynurenic acid (KYN) were used to block the responses of bipolar and horizontal cells. Intravitreal injections of UL-FS 49 or DK-AH 269 were used to block Ih, a hyperpolarization-activated potassium current. Since the microelectrode was referenced to the vitreal electrode, recordings from the inner retina showed only the oscillatory potentials and b-waves. In the inner retina, the potential was flat until the b-wave became measurable, approximately 17 ms from the onset of the flash. The a-wave started to appear as the microelectrode reached the photoreceptors and its amplitude increased with depth until the microelectrode reached the choroid. The a-wave peaked at approximately 8 ms in response to flashes that saturated its amplitude and then began to recover well before any inner retinal responses were apparent. After injections of APB, PDA, and KYN, vitreal and intraretinal recordings showed only the a-wave, which consisted of an increase to peak at approximately 10 ms followed by a recovery to a plateau which was reached at approximately 25 ms. Blockers of Ih reduced the recovery, but did not eliminate it. The a-wave peaks and partially recovers before the b-wave intrudes. Both phases survive blockers of second-order neurons which implies that the photoreceptors generate both the rising and recovery phase of the a-wave. The recovery phase may be due to a current generated by the inner segment of photoreceptors. PMID- 11497413 TI - Effects of directional expectations on motion perception and pursuit eye movements. AB - Expectations about future motions can influence both perceptual judgements and pursuit eye movements. However, it is not known whether these two effects are due to shared processing, or to separate mechanisms with similar properties. We have addressed this question by providing subjects with prior information about the likely direction of motion in an upcoming random-dot motion display and measuring both the perceptual judgements and pursuit eye movements elicited by the stimulus. We quantified the subjects' responses by computing oculometric curves from their pursuit eye movements and psychometric curves from their perceptual decisions. Our results show that directional cues caused similar shifts in both the oculometric and psychometric curves toward the expected motion direction, with little change in the shapes of the curves. Prior information therefore biased the outcome of both eye movement and perceptual decisions without systematically changing their thresholds. We also found that eye movement and perceptual decisions tended to be the same on a trial-by-trial basis, at a higher frequency than would be expected by chance. Furthermore, the effects of prior information were evident during pursuit initiation, as well as during pursuit maintenance, indicating that prior information likely influenced the early processing of visual motion. We conclude that, in our experiments, expectations caused similar effects on both pursuit and perception by altering the activity of visual motion detectors that are read out by both the oculomotor and perceptual systems. Applying cognitive factors such as expectations at relatively early stages of visual processing could act to coordinate the metrics of eye movements with perceptual judgements. PMID- 11497414 TI - Facilitation and suppression of single striate-cell activity by spatially discrete pattern stimuli presented beyond the receptive field. AB - Visual stimulation of a region outside the receptive field of single cells in visual cortex often results in the modulation of their responses. The modulatory effects are thought to be mediated through lateral connections within visual cortex. Research on lateral interactions commonly shows suppression. There has been no systematic study of the optimal conditions for facilitation. Here we have studied the nature of the modulation using a new type of compound stimulus: contrast reversal of pattern stimuli made of three discrete grating patches. The middle patch, optimally fitted to the receptive field in orientation, size, and spatial as well as temporal frequencies, was flanked by two similar patches presented well outside the receptive field. We found that (1) both facilitation and suppression occurred often in the same cells, when orientations of the target and flankers matched the receptive-field's optimal orientation; (2) facilitation with collinear flankers occurred most frequently at target contrasts just above the cell's firing threshold and suppression prevailed at high contrasts; (3) facilitative or suppressive modulation was obtained with target-flankers separation of up to 12 deg or more; (4) collinear facilitation was lost when flankers' orientation was rotated by 90 deg, while keeping all other parameters the same; and (5) neither the modulation mode nor the proportion of modulated cells was related to the cell types (simple vs. complex cells) and cells' laminar locations. Here we have provided physiological evidence for contrast-dependent, collinear facilitation probably underlying perceptual grouping in humans. PMID- 11497415 TI - Salamander UV cone pigment: sequence, expression, and spectral properties. AB - The visual pigment from the ultraviolet (UV) cone photoreceptor of the tiger salamander has been cloned, expressed, and characterized. The cDNA contains a full-length open reading frame encoding 347 amino acids. The phylogenetic analysis indicates that the highest sequence homology is to the visual pigments in the S group. The UV opsin was tagged at the carboxy-terminus with the sequence for the 1D4 epitope. This fusion opsin was expressed in COS-1 cells, regenerated with 11-cis retinal (A1) and immuno-purified, yielding a pigment with an absorbance maximum (lambdamax) of 356 nm which is blue shifted from the absorption of retinal itself. The transducin activation assay demonstrated that this pigment is able to activate rod transducin in a light-dependent manner. Regeneration with 11-cis 3,4-dehydroretinal (A2) yielded a pigment with a lambdamax of 360 nm, only 4 nm red shifted from that of the A1 pigment, while bovine rhodopsin generated with A2 showed a 16-nm red shift from the corresponding A1 pigment. These results demonstrate that the trend for a shorter wavelength pigment to have a smaller shift of lambdamax between the A1 and A2 pigments also fits UV pigments. We hypothesize that the small red shift with A2 could be due to a twist in the chromophore that essentially isolates the ring double bond(s) from conjugation with the rest of the polyene chain. PMID- 11497416 TI - Human neural responses elicited to observing the actions of others. AB - Monkey electrophysiological and human neuroimaging studies indicate the existence of specialized neural systems for the perception and execution of actions. To date, the dynamics of these neural systems in humans have not been well studied. Here, we investigated the spatial and temporal behavior of human neural responses elicited to viewing motion of the face, hand, and body. Scalp event-related potentials (ERPs) were recorded in 20 participants viewing videotaped mouth (opening, closing), hand (closing, opening), and body stepping (forward, backward) movements. ERP peak differences within the movements of each body part were compared using topographical maps of voltage, voltage difference, and Student's t-test at ERP peak latencies. Predominantly temporoparietal negative ERPs occurred to motion of all body parts within 200 ms postmovement onset. Hand closure elicited a significantly greater negativity than opening, particularly in the left hemisphere. Vertex positive ERPs within 300 ms postmovement onset were elicited to hand and body motion. A significantly greater positivity occurred for the body stepping forward relative to stepping backward. The ERP topography was consistent with observed activation foci in human neuroimaging studies. Our data indicate that the neural activity of a system dedicated to the perception of high level motion stimuli can rapidly differentiate between movements across and within body parts. PMID- 11497417 TI - Ocular dominance columns in the adult New World Monkey Callithrix jacchus. AB - In the marmoset Callithrix jacchus, ocular dominance columns (ODC) have been reported to be present in young animals, but absent in adults (Spatz, 1989). We have studied in juvenile and adult animals the postnatal organization of the retino-geniculo-cortical afferents by means of transneuronal labeling. We show in the present work that ODC are present in the primary visual cortex of Callithrix jacchus, both in the adult and in the juvenile animal. The present work confirms the presence of ODC in the visual cortex of juvenile marmoset before the end of the first postnatal month. In 2-month-old animals, ODC are well demarcated in IVcalpha and IVcbeta. In the adult marmosets, the present data clearly show that the primary visual cortex is also organized with ODC. In horizontal sections, they form a mosaic through the ventral and dorsal calcarine cortex and through the dorso-lateral occipital part of the striate cortex. In frontal sections, their presence is manifest in IVcbeta within the calcarine cortex and they only faintly appear in IVcalpha. These new findings are important since they underline the usefulness of the adult New World Monkeys as a model in visual research. PMID- 11497418 TI - Localization of amino acid neurotransmitters following in vitro ischemia and anoxia in the rat retina. AB - Glutamate and gamma-aminobutyric acid (GABA) are two of the dominant neurotransmitters in the retina and brain. The production/degradation of glutamate and GABA involves an intricate interrelationship between neurons and glia, as well as aerobic and anaerobic metabolic pathways. The aim of this work was to develop an in vitro model of retinal ischemia/anoxia and determine the changes in cellular localization of glutamate and GABA and the time course for such changes. After anoxic/ischemic insult, glutamate and GABA rapidly accumulate within glia with GABA showing a quicker time course and larger magnitude change. The accumulation time constant for both glutamate and GABA under anoxic conditions was dependent upon glucose concentration: high glucose levels resulted in delayed glial amino acid loading. The differences in time constants between GABA and glutamate glial loading most likely reflect the multitude of glutamate degradation pathways compared to the single aerobically dependent GABA pathway. Oxygen availability and reduced glucose (hypoglycemia) lead to an almost immediate increase (within 1 min) of glutamate and GABA labelling within glia. In addition, altered labelling patterns were found under anoxic/ischemic conditions for amino acids involved in glutamate transamination reactions: aspartate, leucine, alanine. and ornithine. These changes are consistent with alterations of equilibria of enzymatic reactions involved in glutamate metabolism, and thus support a role for all four amino acids in glutamate metabolism within a variety of retinal neurons. PMID- 11497419 TI - Heat shock protein 90 in retinal ganglion cells: association with axonally transported proteins. AB - The mRNAs for heat shock protein 90 (HSP90) are found at highest levels (differentially expressed) in the primate retinal fovea, the region of highest visual acuity, compared to the peripheral retina. HSP90 expression and retinal associations were analyzed by immuno-localization, in situ hybridization, and western analysis. Retinal ganglion cells (RGCs) express much of the HSP90 mRNA present in the primate retinal fovea. A large fraction of RGC synthesized HSP90 is apparently present in the axonal compartment. To identify the role of HSP90 protein in the optic nerve and retina, co-immunoprecipitation experiments were performed, using antibodies specific for HSP90 isoforms. The immunoprecipitates were analyzed for neurotrophin receptor and ligand activities, and MAP kinase activity. MAP kinase assay was used to determine the activation state of MAP kinase associated with HSP90. HSP90 proteins selectively associate with the inactive form of full-length tyrosine kinase growth factor receptor trkB, suggesting utilization during anterograde axonal transport. Activated MAP kinase, associated with the trk downstream signaling cascade, was found to co immunoprecipitate with optic nerve HSP90, suggesting that HSP90 may be utilized in retrograde transport of the secondary messengers associated with neurotrophin signaling. HSP90 can thus be hypothesized to play a role in bidirectional RGC axonal protein transport. PMID- 11497420 TI - Comparative structural and functional analysis of photoreceptor neurons of Rho-/- mice reveal increased survival on C57BL/6J in comparison to 129Sv genetic background. AB - To explore the possible influence of defined genetic backgrounds on photoreceptor viability and function in mice carrying a targeted disruption of the rhodopsin gene, the severities of retinopathies in Rho-/- mice on C57BL/6J and 129Sv congenic backgrounds were compared by light microscopy and electroretinography and qualitatively by in situ end labeling of DNA in apoptotic photoreceptor nuclei of retinal sections. Cone photoreceptor viability and function were shown to deteriorate more slowly on the C57BL/6J background in comparison to that of the 129Sv, with significantly greater numbers of outer nuclear layer nuclei in the retinas of C57BL/6J mice at 3 and 4 months of age. Both amplitude and waveform features of the ERG were shown to be remarkably different in the two strains, indicating an approximately 6-fold difference in C57BL/6J Rho-/- mice compared to 129Sv Rho-/- mice at 80 days. Thus, in comparison with the 129Sv strain, genetic modifiers appear to constitute a component of the C57BL/6J background, the expression of which significantly protects cone photoreceptors from apoptotic death in a mutation-induced murine retinopathy. The differences in phenotype revealed in this study are sufficient in principle to provide a basis for comparisons to be made between QTLs in light-induced and mutation-induced systems. PMID- 11497421 TI - Characterization of the rod photoresponse isolated from the dark-adapted primate ERG. AB - The a-wave of the human dark-adapted ERG is thought to derive from activity of rod photoreceptors. However, other sources within the retina could potentially perturb this simple equation. We investigated the extent to which the short latency dark-adapted rod a-wave of the primate ERG is dominated by the rod photoresponse and the applicability of the phototransduction model to fit the rod a-wave. Dark-adapted Ganzfeld ERGs were elicited over a 5-log-unit intensity range using short bright xenon flashes, and the light-adapted cone responses were subtracted to isolate the rod ERG a-wave. Intravitreal 4-phosphono-butyric acid (APB) and cis-2,3-piperidine-dicarboxylic acid (PDA) were applied to isolate the photoreceptor response. The Hood and Birch version of the phototransduction model, Rmax[1 - e(-I x S x (t-t(eff)))2], was fitted to the a-wave data while allowing Rmax and S to vary. Three principle observations were made: (1) At flash intensities > or =0.77 log sc-td-s the leading edge of the normalized rod ERG a wave tracks the isolated photoreceptor response across the first 20 ms or up to the point of b-wave intrusion. The rod ERG a-wave was essentially identical to the isolated receptor response for all intensities that produce peak responses within 14 ms after the flash. (2) The best fit of sensitivity (S) was not affected by APB and/or PDA, suggesting that the inner retina contributes very little to the dark-adapted a-wave. (3) APB always reduced the maximum dark adapted a-wave amplitude (by 15-30%), and PDA always increased it (by 7-15%). Using the phototransduction model, both events can be interpreted as a scaling of the photoreceptor dark current. This suggests that activity of postreceptor cells somehow influences the rod dark current, possibly by feedback through horizontal cells (although currently not demonstrated for the rod system), or by altering the ionic concentrations near the photoreceptors, or by neuromodulator effects mediated by dopamine or melatonin. PMID- 11497422 TI - GABAergic visual pathways in the frog Rana pipiens. AB - Gamma-aminobutyric acid (GABA) is the most prevalent inhibitory neurotransmitter in the vertebrate brain. It can exert its influence either as GABAergic projection pathways or as local interneurons, which play an essential role in many visual functions. However, no GABAergic visual pathways have been studied in frogs so far. In the present study, GABAergic pathways in the central visual system of Rana pipiens were investigated with double-labeling techniques, combining immunocytochemistry for GABA with Rhodamine microspheres for retrograde tracing. Three GABAergic visual pathways were identified: (1) a retino-tectal projection, from retina to the contralateral optic tectum (OT); (2) an ipsilateral projection from the nucleus of the basal optic root (nBOR) to the pretectal nucleus lentiformis mesencephali (nLM); and (3) a second-order pathway from the nucleus isthmi (NI), bilaterally, to the optic tectum. These results indicate that GABA is involved in both first-order (retina to optic tectum) as well as second-order (nucleus isthmi to optic tectum) visual projections in Rana pipiens, and may play a major role in mediating visuomotor reflexs such as optokinetic nystagmus or other visually guided behaviors. PMID- 11497423 TI - Linear filtering and nonlinear interactions in direction-selective visual cortex neurons: a noise correlation analysis. AB - Spatial and temporal properties related to direction selectivity of both simple and complex type visual cortex neurons were assessed by cross-correlation analysis of their responses to random ternary white noise. This stimulus consisted of multiple randomly placed bars, each colored white, black, or gray with equal probability, which were rerandomized every 5-10 ms. A first-order cross-correlation analysis of a neuron's spike train with the spatiotemporal history of the stimulus provided an estimate of the neuron's linear spatiotemporal filtering properties. A nonlinear correlation analysis measured the amount of interaction for pair-wise combinations of bars as a function of their relative spatial and temporal separations. The spatiotemporal orientation of each of these functions was quantified using a "motion energy index" (MEI), which was compared to the neurons' direction selectivity measured with drifting sinewave gratings. Both first-order and nonlinear correlation plots usually showed s-t orientation whose sign was consistent with the neuron's direction preference; however, in many cases the MEI for first-order analysis was weak compared to that seen in the nonlinear interactions. The structures of the nonlinear interaction functions were also compared with predictions from a conventional model of direction selectivity based on a simple spatiotemporally oriented linear filter, followed by an intensive nonlinearity ("LN model"). These comparisons showed that some neurons' data agreed reasonably well with such a model, while others agreed poorly or not at all. Simulations of an alternative model which combines signals from idealized lagged and nonlagged front-end linear filters produce noise correlation results more like those seen in the neurophysiological data. PMID- 11497424 TI - Task-specific reversal of visual hemineglect following bilateral reversible deactivation of posterior parietal cortex: a comparison with deactivation of the superior colliculus. AB - The purpose of the present study was to compare and contrast behavioral performance on three different tasks of spatial cognition during unilateral and bilateral reversible deactivation of posterior parietal cortex. Specifically, we examined posterior middle suprasylvian (pMS) sulcal cortex in adult cats during temporary and reversible cooling deactivation. In Task 1, the cats oriented to a high-contrast, black visual stimulus moved into the visual field periphery. In Task 2, the cats oriented to a static light-emitting diode (LED). Task 3 examined the cats' ability to determine whether a black-and-white checkered, landmark box was closer to the right or left side of the testing apparatus. Following training on all tasks, cryoloops were implanted bilaterally within the pMS sulcus. Unilateral deactivation of pMS sulcal cortex resulted in virtually no responses to either moved or static stimuli and virtually no responses to landmarks presented in the contralateral hemifield, and a profound contralateral hemifield neglect was induced. Responses to stimuli and landmarks presented in the ipsilateral hemifield were unimpaired. Additive, bilateral cooling of the homotopic region in the contralateral hemisphere, but not an adjacent region, resulted in reversal of the initial hemineglect for the moved stimulus, yet induced a complete failure to orient to peripheral static LED stimuli. Bilateral cooling also reversed the contralateral neglect of the landmark, but then cats could not accurately determine position of the landmark anywhere in the visual field because performance was reduced to chance levels for all landmark loci in both hemifields. In this instance, as the contralateral neglect disappeared during bilateral cooling of pMS cortex, a new spatial discrimination deficit was revealed across the entire visual field. We conclude that pMS cortex contributes in multiple ways to the analyses of space, and that these contributions cannot be safely predicted from analyses of unilateral deactivations or from one task to another. Moreover, it is clear that other structures are capable of guiding orienting to high contrast, moved targets when pMS cortex is eliminated from brain circuitry. However, these same structures are incapable of supporting either orienting to static stimuli or analyses of spatial relations as tested with the landmark task. The impact of reversible deactivation of the superior colliculus on these same tasks is discussed. PMID- 11497425 TI - Translation: in retrospect and prospect. AB - This review is occasioned by the fact that the problem of translation, which has simmered on the biological sidelines for the last 40 years, is about to erupt center stage--thanks to the recent spectacular advances in ribosome structure. This most complex, beautiful, and fascinating of cellular mechanisms, the translation apparatus, is also the most important. Translation not only defines gene expression, but it is the sine qua non without which modern (protein-based) cells would not have come into existence. Yet from the start, the problem of translation has been misunderstood--a reflection of the molecular perspective that dominated Biology of the last century. In that the our conception of translation will play a significant role in creating the structure that is 21st century Biology, it is critical that our current (and fundamentally flawed) view of translation be understood for what it is and be reformulated to become an all embracing perspective about which 21st century Biology can develop. Therefore, the present review is both a retrospective and a plea to biologists to establish a new evolutionary, RNA-World-centered concept of translation. What is needed is an evolutionarily oriented perspective that, first and foremost, focuses on the nature (and origin) of a primitive translation apparatus, the apparatus that transformed an ancient evolutionary era of nucleic acid life, the RNA World, into the world of modern cells. PMID- 11497426 TI - The La antigen associates with the human telomerase ribonucleoprotein and influences telomere length in vivo. AB - La is an important component of ribonucleoprotein complexes and telomerase is a ribonucleoprotein that compensates for the shortening of the ends of linear DNA by adding telomeric repeats onto the ends of chromosomes by using an integral RNA as the template. We have identified a direct and specific interaction between La and the RNA component of human telomerase. Antibodies specific to La precipitate the human telomerase ribonucleoprotein complex derived from tumor cells, telomerase immortalized normal cells, and in vitro transformed cells. Overexpression of La in both experimentally immortalized human cells and prostate cancer cells results in gradual telomere shortening. Our results demonstrate that La can associate with telomerase and its expression level can influence telomere homeostasis in vivo. PMID- 11497427 TI - Staphylococcus aureus domain V functions in Escherichia coli ribosomes provided a conserved interaction with domain IV is restored. AB - Domain V of Escherichia coli 23 S rRNA (residues 2023-2630) was replaced by that from Staphylococcus aureus, thereby introducing 132 changes in the rRNA sequence. The resulting ribosomal mutant was unable to support cell growth. The mutant was rescued, however, by restoring an interaction between domains IV and V (residues 1782 and 2586). Although the importance of this interaction, U/U in E. coli, C/C in S. aureus, is therefore demonstrated, it cannot be the only tertiary interaction important for ribosomal function as the rescued hybrid grew more slowly than the wild type. Additionally, although the single-site mutations U1782C and U2586C in E. coli are viable, the double mutant is lethal. PMID- 11497428 TI - Ribosomal protein L5 helps anchor peptidyl-tRNA to the P-site in Saccharomyces cerevisiae. AB - Our previous demonstration that mutants of 5S rRNA called mof9 can specifically alter efficiencies of programmed ribosomal frameshifting (PRF) suggested a role for this ubiquitous molecule in the maintenance of translational reading frame, though the repetitive nature of the 5S rDNA gene (>100 copies/cell) inhibited more detailed analyses. However, given the known interactions between 5S rRNA and ribosomal protein L5 (previously called L1 or YL3) encoded by an essential, single-copy gene, we monitored the effects of a series of well-defined rpl5 mutants on PRF and virus propagation. Consistent with the mof9 results, we find that the rpl5 mutants promoted increased frameshifting efficiencies in both the 1 and +1 directions, and conferred defects in the ability of cells to propagate two endogenous viruses. Biochemical analyses demonstrated that mutant ribosomes had decreased affinities for peptidyl-tRNA. Pharmacological studies showed that sparsomycin, a peptidyltransferase inhibitor that specifically increases the binding of peptidyl-tRNA with ribosomes, was antagonistic to the frameshifting defects of the most severe mutant, and the extent of sparsomycin resistance correlated with the severity of the frameshifting defects in all of the mutants. These results provide biochemical and physiological evidence that one function of L5 is to anchor peptidyl-tRNA to the P-site. A model is presented describing how decreased affinity of ribosomes for peptidyl-tRNA can affect both -1 and +1 frameshifting, and for the effects of sparsomycin. PMID- 11497429 TI - Structural features in the HIV-1 repeat region facilitate strand transfer during reverse transcription. AB - Two obligatory DNA strand transfers take place during reverse transcription of a retroviral RNA genome. The first strand transfer is facilitated by terminal repeat (R) elements in the viral genome. This strand-transfer reaction depends on base pairing between the cDNA of the 5'R and the 3'R. There is accumulating evidence that retroviral R regions contain features other than sequence complementarity that stimulate this critical nucleic acid hybridization step. The R region of the human immunodeficiency virus type 1 (HIV-1) is relatively extended (97 nt) and encodes two well-conserved stem-loop structures, the TAR and poly(A) hairpins. The role of these motifs was studied in an in vitro strand transfer assay with two separate templates, the 5'R donor and the 3'R acceptor, and mutants thereof. The results indicate that the upper part of the TAR hairpin structure in the 5'R donor is critical for efficient strand transfer. This seems to pose a paradox, as the 5'R template is degraded by RNase H before strand transfer occurs. We propose that it is not the RNA hairpin motif in the 5'R donor, but rather the antisense motif in the ssDNA copy, which can also fold a hairpin structure, that is critical for strand transfer. Mutation of the loop sequence in the TAR hairpin of the donor RNA, which is copied in the loop of the cDNA hairpin, reduces the transfer efficiency more than fivefold. It is proposed that the natural strand-transfer reaction is enhanced by interaction of the anti TAR ssDNA hairpin with the TAR hairpin in the 3'R acceptor. Base pairing can occur between the complementary loops ("loop-loop kissing"), and strand transfer is completed by the subsequent formation of an extended RNA-cDNA duplex. PMID- 11497430 TI - Specific metal-ion binding sites in a model of the P4-P6 triple-helical domain of a group I intron. AB - Divalent metal ions play a crucial role in RNA structure and catalysis. Phosphorothioate substitution and manganese rescue experiments can reveal phosphate oxygens interacting specifically with magnesium ions essential for structure and/or activity. In this study, phosphorothioate interference experiments in combination with structural sensitive circular dichroism spectroscopy have been used to probe molecular interactions underlying an important RNA structural motif. We have studied a synthetic model of the P4-P6 triple-helical domain in the bacteriophage T4 nrdB group I intron, which has a core sequence analogous to the Tetrahymena ribozyme. Rp and Sp sulfur substitutions were introduced into two adjacent nucleotides positioned at the 3' end of helix P6 (U452) and in the joining region J6/7 (U453). The effects of sulfur substitution on triple helix formation in the presence of different ratios of magnesium and manganese were studied by the use of difference circular dichroism spectroscopy. The results show that the pro-Sp oxygen of U452 acts as a ligand for a structurally important magnesium ion, whereas no such effect is seen for the pro-Rp oxygen of U452. The importance of the pro-Rp and pro-Sp oxygens of U453 is less clear, because addition of manganese could not significantly restore the triple-helical interactions within the isolated substituted model systems. The interpretation is that U453 is so sensitive to structural disturbance that any change at this position hinders the proper formation of the triple helix. PMID- 11497431 TI - Poly(rC) binding proteins mediate poliovirus mRNA stability. AB - The 5'-terminal 88 nt of poliovirus RNA fold into a cloverleaf RNA structure and form ribonucleoprotein complexes with poly(rC) binding proteins (PCBPs; AV Gamarnik, R Andino, RNA, 1997, 3:882-892; TB Parsley, JS Towner, LB Blyn, E Ehrenfeld, BL Semler, RNA, 1997, 3:1124-1134). To determine the functional role of these ribonucleoprotein complexes in poliovirus replication, HeLa S10 translation-replication reactions were used to quantitatively assay poliovirus mRNA stability, poliovirus mRNA translation, and poliovirus negative-strand RNA synthesis. Ribohomopoly(C) RNA competitor rendered wild-type poliovirus mRNA unstable in these reactions. A 5'-terminal 7-methylguanosine cap prevented the degradation of wild-type poliovirus mRNA in the presence of ribohomopoly(C) competitor. Ribohomopoly(A), -(G), and -(U) did not adversely affect poliovirus mRNA stability. Ribohomopoly(C) competitor RNA inhibited the translation of poliovirus mRNA but did not inhibit poliovirus negative-strand RNA synthesis when poliovirus replication proteins were provided in trans using a chimeric helper mRNA possessing the hepatitis C virus IRES. A C24A mutation prevented UV crosslinking of PCBPs to 5' cloverleaf RNA and rendered poliovirus mRNA unstable. A 5'-terminal 7-methylguanosine cap blocked the degradation of C24A mutant poliovirus mRNA. The C24A mutation did not inhibit the translation of poliovirus mRNA nor diminish viral negative-strand RNA synthesis relative to wild-type RNA. These data support the conclusion that poly(rC) binding protein(s) mediate the stability of poliovirus mRNA by binding to the 5'-terminal cloverleaf structure of poliovirus mRNA. Because of the general conservation of 5' cloverleaf RNA sequences among picornaviruses, including C24 in loop b of the cloverleaf, we suggest that viral mRNA stability of polioviruses, coxsackieviruses, echoviruses, and rhinoviruses is mediated by interactions between PCBPs and 5' cloverleaf RNA. PMID- 11497432 TI - Coevolution of group II intron RNA structures with their intron-encoded reverse transcriptases. AB - Catalytic RNAs are often regarded as molecular fossils from the RNA World, yet it is usually difficult to get more specific information about their evolution. Here we have investigated the coevolution of group II intron RNA structures with their intron-encoded reverse transcriptases (RTs). Unlike group I introns, there has been no obvious reshuffling between intron RNA structures and ORFs. Of the six classes of intron structures that encode ORFs, three are conventional forms of group II A1, B1, and B2 secondary structures, whereas the remaining classes are bacterial, are possibly associated with the most primitive ORFs, and have unusual features and hybrid features of group IIA and group IIB intron structures. Based on these data, we propose a new model for the evolution of group II introns, designated the retroelement ancestor hypothesis, which predicts that the major RNA structural forms of group II introns developed through coevolution with the intron-encoded protein rather than as independent catalytic RNAs, and that most ORF-less introns are derivatives of ORF-containing introns. The model is supported by the distribution of ORF-containing and ORF-less introns, and by numerous examples of ORF-less introns that contain ORF remnants. PMID- 11497434 TI - The GA motif: an RNA element common to bacterial antitermination systems, rRNA, and eukaryotic RNAs. AB - Two different transcription termination control mechanisms, the T box and S box systems, are used to regulate transcription of many bacterial aminoacyl-tRNA synthetase, amino acid biosynthesis, and amino acid transport genes. Both of these regulatory mechanisms involve an untranslated mRNA leader region capable of adopting alternate structural conformations that result in transcription termination or transcription elongation into the downstream region. Comparative analyses revealed a small RNA secondary structural element, designated the GA motif, that is highly conserved in both T box and S box leader sequences. The motif consists of two short helices separated by an asymmetric internal loop, with highly conserved GA dinucleotide sequences on either side of the internal loop. Site-directed mutagenesis of this motif in model T and S box leader sequences indicated that it is essential for transcriptional regulation in both systems. This motif is similar to the binding site of yeast ribosomal protein L30, the Snu13p binding sites found in U4 snRNA and box C/D snoRNAs, and two elements in 23S rRNA. PMID- 11497433 TI - Function and subnuclear distribution of Rpp21, a protein subunit of the human ribonucleoprotein ribonuclease P. AB - Rpp21, a protein subunit of human nuclear ribonuclease P (RNase P) was cloned by virtue of its homology with Rpr2p, an essential subunit of Saccharomyces cerevisiae nuclear RNase P. Rpp21 is encoded by a gene that resides in the class I gene cluster of the major histocompatibility complex, is associated with highly purified RNase P, and binds precursor tRNA. Rpp21 is predominantly localized in the nucleoplasm but is also observed in nucleoli and Cajal bodies when expressed at high levels. Intron retention and splice-site selection in Rpp21 precursor mRNA regulate the intranuclear distribution of the protein products and their association with the RNase P holoenzyme. Our study reveals that dynamic nuclear structures that include nucleoli, the perinucleolar compartment and Cajal bodies are all involved in the production and assembly of human RNase P. PMID- 11497435 TI - Initiation factor IF 2 binds to the alpha-sarcin loop and helix 89 of Escherichia coli 23S ribosomal RNA. AB - During initiation of protein synthesis in bacteria, translation initiation factor IF2 is responsible for the recognition of the initiator tRNA (fMet-tRNA). To perform this function, IF2 binds to the ribosome interacting with both 30S and 50S ribosomal subunits. Here we report the topographical localization of translation initiation factor IF2 on the 70S ribosome determined by base-specific chemical probing. Our results indicate that IF2 specifically protects from chemical modification two sites in domain V of 23S rRNA, namely A2476 and A2478, and residues around position 2660 in domain VI, the so-called sarcin-ricin loop. These footprints are generated by IF2 regardless of the presence of fMet-tRNA, GTP, mRNA, and IF1. IF2 causes no specific protection of 16S rRNA. We observe a decreased reactivity of residues A1418 and A1483, which is an indication that the initiation factor has a tightening effect on the association of ribosomal subunits. This result, confirmed by sucrose density gradient analysis, seems to be a universally conserved property of IF2. PMID- 11497436 TI - Seasonal changes in serum calcium, phosphorus, and vitamin D concentrations in llamas and alpacas. AB - OBJECTIVE: To evaluate the interaction of season and age on serum calcium, phosphorus, and vitamin D3 concentrations in llamas and alpacas. ANIMALS: 23 clinically normal llamas and 7 alpacas. PROCEDURES: Animals were assigned to 1 of the 3 following groups on the basis of age at the start of the study: adult (age, > or = 24 months; n = 8), yearling (> 12 but < 20 months; 5), and neonate (< 6 months; 17). Twelve serum samples were obtained at monthly intervals. Calcium, phosphorus, and vitamin D3 concentrations were measured, and the calcium-to phosphorus concentration (Ca:P) ratio calculated. Effect of season and age on each of these variables was determined. RESULTS: Vitamin D3 concentrations varied significantly as a function of season; the highest and lowest concentrations were detected September through October and February through March, respectively. The seasonal decrease in vitamin D3 concentration was significantly greater in neonates and yearlings, compared with adults. Serum phosphorus concentration decreased as a function of age, with the most significant seasonal change detected in the neonate group. The Ca:P ratio in neonates varied between 1.1 and 1.3 except during winter months when it increased to > or = 2.0. CONCLUSIONS AND CLINICAL RELEVANCE: Mean vitamin D3 concentration varied by > 6 fold in neonatal and yearling llamas and alpacas and > 3 fold in adult animals as a function of season. These results support the hypothesis that seasonal alterations in vitamin D3 concentrations are a key factor in the development of hypophosphatemic rickets in llamas and alpacas. PMID- 11497437 TI - Rapid infection in market-weight swine following exposure to a Salmonella typhimurium-contaminated environment. AB - OBJECTIVE: To evaluate the possibility of swine becoming infected with Salmonella Typhimurium when housed for 2 to 6 hours in an environment contaminated with Salmonella, similar to a lairage situation prior to slaughter. ANIMALS: 40 crossbred market pigs with an approximate body weight of 92 kg. PROCEDURE: Five trials were conducted (8 pigs/trial) in simulated lairage conditions. Superficial inguinal, ileocecal, and mandibular lymph nodes, cecal contents, distal portion of the ileum, and fecal samples were obtained from each pig after 2 (n = 10), 3 (10), and 6 (5) hours of exposure to an environment contaminated with feces defecated by 10 pigs intranasally inoculated with nalidixic acid-resistant Salmonella Typhimurium (chi4232). In addition, 5 control pigs that were not exposed were also evaluated in the same manner. RESULTS: Feces deposited on the floor by intranasally inoculated swine were mixed with water to form slurry with a resulting load of approximately 10(3) colony-forming units of Salmonella Typhimurium/g of material. Eight of 10, 6 of 10, and 6 of 6 pigs exposed to the slurry for 2, 3, or 6 hours, respectively, had positive results for at least 1 sample when tested for the specific strain of Salmonella Typhimurium. CONCLUSIONS AND CLINICAL RELEVANCE: Pigs can become infected during routine resting or holding periods during marketing when exposed to relatively low amounts of Salmonella organisms in the preslaughter environment. Intervention at this step of the production process may have a major impact on the safety of pork products. PMID- 11497438 TI - Heritability estimations for diseases, coat color, body weight, and height in a birth cohort of Boxers. AB - OBJECTIVE: To obtain heritability estimates for diseases and characteristics in Boxers. ANIMALS: Birth cohort of 2,929 purebred Boxers from 414 litters. PROCEDURE: Heritability estimates were determined for cheiloschisis palatoschisis, cryptorchidism, epilepsy, stifle disorders, cardiac disorders, coat color, birth weight, and adult weight, and height. Binary traits were analyzed by use of a mixed-effects probit model. Some traits also were analyzed by use of a model that postulated monogenic inheritance. Full pedigree analyses were performed. Variation in incidences of disease among clusters of related dogs was evaluated. RESULTS: Heritability estimates were virtually zero for cardiac disorders, medium (0.17 to 0.36) for most other traits, and high (> 0.55) for coat color, birth weight, and adult height. Litter effects and risk factors affected cheiloschisis-palatoschisis, heart murmur, coat color, broadly defined epilepsy, and adult weight. Litter effects may be attributable to common environmental effects for littermates but also may be attributable to dominance variation caused by a recessive gene. Heritability estimates increased when stricter definitions for epilepsy and stifle disorders were used. The monogenic model did not reveal higher heritability estimates for 6 traits analyzed. Incidences for white coat differed significantly for 10 familial clusters, confirming high heritability and effects of familial lineage. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that genetic improvement of most traits should be feasible, except for cardiac disorders. However, because most traits are influenced by environmental effects as well as genetic effects, genetic counseling based on polygenic inheritance and use of familial information rather than strict exclusion of parents is preferred. PMID- 11497439 TI - Long-term temporal evaluation of ground reaction forces during development of experimentally induced osteoarthritis in dogs. AB - OBJECTIVE: To describe changes in vertical ground reaction forces (GRF) over 48 months in dogs with osteoarthritis (OA) of the stifle joint induced by transection of a cranial cruciate ligament (CCL). ANIMALS: 12 clinically normal adult dogs. PROCEDURE: Vertical GRF (eg, peak force and impulse) were determined prior to and 1, 2, 3, 6, 10, and 12 months after transection of the right CCL. In 7 dogs, data were also collected 24, 32, 38, 42, and 48 months after transection. RESULTS: Vertical peak force and impulse were significantly decreased in the right hind limb at all times after transection, compared with baseline values. From 10 through 48 months after transection, vertical GRF remained essentially static. Ground reaction forces in the unoperated (left) hind limb also changed significantly during the study. Left vertical impulse significantly increased 3 months after transection, whereas at 24, 38, 42, and 48 months after transection, left vertical peak force was significantly decreased, compared with the baseline value. Mean intradog coefficients of variation (CV) for peak vertical force and impulse ranged from 738 and 9.32, respectively, 1 month after transection to 1.96 and 2.76, respectively, at 42 months. CONCLUSIONS AND CLINICAL RELEVANCE: Vertical GRF in the affected hind limb equilibrated approximately 10 months after CCL transection. Prior to this, force transmission across the affected stifle joint changed significantly over time. Intradog CV were small, indicating that GRF may be an appropriate outcome measurement for evaluation of OA development induced by CCL transection in dogs. PMID- 11497440 TI - Expression of a transforming gene (E5) of bovine papillomavirus in sarcoids obtained from horses. AB - OBJECTIVE: To determine expression of a transforming gene (E5) of bovine papillomavirus in sarcoids, other tumors, and normal skin samples collected from horses with and without sarcoids. SAMPLE POPULATION: 23 sarcoids and 6 samples of normal skin obtained from 16 horses with sarcoids, 2 samples of normal skin and 2 papillomas obtained from horses without sarcoids, and 1 papilloma obtained from a cow. PROCEDURE: Protein was extracted from tissue samples collected from horses and incubated with agarose beads covalently coupled to Staphylococcus aureus protein A and an anti-E5 polyclonal antibody. Following incubation, proteins were eluted from the beads and electrophoresed on a 14% polyacrylamide gel and transferred to a polyvinylidene difluoride membrane. The E5 protein was detected by use of western blot analysis, using a chemiluminescence detection system. RESULTS: All 23 sarcoids had positive results for expression of E5 protein. Quantity of viral protein appeared to vary among sarcoids. All other tissues examined had negative results for E5 protein. Highest expression for E5 protein was observed in biologically aggressive fibroblastic variants of sarcoids, compared with expression in quiescent tumors. CONCLUSIONS AND CLINICAL RELEVANCE: This study documented that activation and expression of the E5 gene is evident in sarcoids obtained from horses. These data support the conclusion that infection with bovine papillomavirus is important in the initiation or progression of sarcoids in horses. Treatment strategies designed to increase immune recognition of virally infected cells are warranted. PMID- 11497441 TI - Effects of dexamethasone on emesis in cats sedated with xylazine hydrochloride. AB - OBJECTIVE: To determine antiemetic efficacy of prophylactic administration of dexamethasone and its influence on sedation in cats sedated with xylazine hydrochloride. ANIMALS: 6 healthy adult cats (3 males and 3 females). PROCEDURE: The prophylactic antiemetic effect of 4 doses of dexamethasone (1, 2, 4, and 8 mg/kg of body weight, IM) or saline (0.9% NaCl) solution (0.066 ml/kg, IM) administered 1 hour before administration of xylazine (0.66 mg/kg, IM) was evaluated. Cats initially were given saline treatment (day 0) and were given sequentially increasing doses of xylazine on days 7, 14, 21, and 28. After xylazine injection, all cats were observed for 30 minutes to allow assessment of frequency of emesis and time until onset of the first emetic episode. The influence of dexamethasone on xylazine-induced sedation in these cats also was evaluated. RESULTS: Prior treatment with 4 or 8 mg/kg of dexamethasone significantly reduced the frequency of emetic episodes and also significantly prolonged the time until onset of the first emetic episode after xylazine injection. Time until onset of the first emetic episode also was significantly prolonged for dexamethasone at a dose of 2 mg/kg. Time until onset of sedation after administration of xylazine was not altered by administration of dexamethasone. CONCLUSIONS AND CLINICAL RELEVANCE: Dexamethasone (4 or 8 mg/kg, IM) significantly decreased the frequency of emetic episodes induced by xylazine without compromising sedative effects in cats. Dexamethasone may be used prophylactically as an antiemetic in cats treated with xylazine. PMID- 11497442 TI - Cloning and characterization of a Moraxella bovis cytotoxin gene. AB - OBJECTIVE: To identify the Moraxella bovis cytotoxin gene. PROCEDURE: Hemolytic and nonhemolytic strains of M. bovis were compared by use of western blotting to identify proteins unique to hemolytic strains. Oligonucleotide primers, designed on the basis of amino acid sequences of 2 tryptic peptides derived from 1 such protein and conserved regions of the C and B genes from members of the repeats in the structural toxin (RTX) family of bacterial toxins, were used to amplify cytotoxin-specific genes from M. bovis genomic DNA. Recombinant proteins were expressed, and antisera against these proteins were produced in rabbits. RESULTS: Several proteins ranging in molecular mass from 55 to 75 kd were unique to the hemolytic strain. An open reading frame encoding a 927-amino acid protein with a predicted molecular mass of 98.8 kd was amplified from M. bovis genomic DNA. The deduced amino acid sequence encoded by this open reading frame was homologous to RTX toxins. Antisera against the recombinant carboxy terminus encoded by this open reading frame neutralized hemolytic and cytolytic activities of native M. bovis cytotoxin. CONCLUSIONS AND CLINICAL RELEVANCE: A gene was identified in M bovis that encodes a protein with sequence homology to other RTX toxins. Results of cytotoxin neutralization assays support the hypothesis that M. bovis cytotoxin is encoded by this gene and belongs in the RTX family of bacterial exoproteins. Identification of this gene and expression of recombinant cytotoxin could facilitate the development of improved vaccines against infectious bovine keratoconjunctivitis. PMID- 11497443 TI - Evaluation of monoclonal antibodies for identification of subpopulations of myeloid cells in bone marrow obtained from dogs. AB - OBJECTIVE: To evaluate monoclonal antibodies that may be useful for immunophenotyping myeloid cells in bone marrow of dogs. SAMPLE POPULATION: Bone marrow specimens obtained from 5 dogs. DESIGN: Specimens were labeled with monoclonal antibodies that detected CD18, major histocompatability antigen class II (MHC class-II), CD14, and Thy-1. Cells labeled with each of the antibodies were isolated by use of a fluorescence-activated cell sorter. Differential cell counts of sorted cells were used to determine cells that were labeled by each of the various antibodies. RESULTS: Myeloid cells labeled with anti-CD18 antibody included granulocytes, lymphocytes, and monocytes-macrophages. Immature and mature granulocytes were labeled. Lymphocytes, monocytes-macrophages, and eosinophils were labeled with anti-Thy-1 antibody. Cells labeled with anti-MHC class II antibody included approximately 9% of bone marrow cells, which consisted almost exclusively of lymphocytes and monocytes-macrophages. Approximately 4% of bone marrow cells were labeled with anti-CD14 antibody, with > 90% of sorted cells being monocytes-macrophages. CONCLUSIONS AND CLINICAL RELEVANCE: Four monoclonal antibodies for use in detecting subpopulations of canine bone marrow cells were evaluated. These antibodies should be useful in differentiating the origin of leukemic cells in dogs. PMID- 11497444 TI - Metabolic alterations in dogs with osteosarcoma. AB - OBJECTIVE: To evaluate changes in resting energy expenditure (REE) as well as protein and carbohydrate metabolism in dogs with osteosarcoma (OSA). ANIMALS: 15 weight-stable dogs with OSA that did not have other concurrent metabolic or endocrine illness and twelve 1-year-old sexually intact female Beagles (control dogs). PROCEDURES: Indirect calorimetry was performed on all dogs to determine REE and respiratory quotient (RQ). Stable isotope tracers (15N-glycine, 4.5 mg/kg of body weight, IV; 6,6-deuterium-glucose, 4.5 mg/kg, IV as a bolus, followed by continuous-rate infusion at 1.5 mg/kg/h for 3 hours) were used to determine rate of protein synthesis and glucose flux in all dogs. Dual-energy x-ray absorptiometry (DEXA) scans were performed to determine total body composition. RESULTS: Accounting for metabolic body size, REE in dogs with OSA was significantly higher before and after surgery, compared with REE of healthy control dogs. The RQ values did not differ significantly between groups. Dogs with OSA also had decreased rates of protein synthesis, increased urinary nitrogen loss, and increased glucose flux during the postoperative period. CONCLUSIONS AND CLINICAL RELEVANCE: Alterations in energy expenditure, protein synthesis, urinary nitrogen loss, and carbohydrate flux were evident in dogs with OSA, similar to results documented in humans with neoplasia. Changes were documented in REE as well as protein and carbohydrate metabolism in dogs with OSA. These changes were evident even in dogs that did not have clinical signs of cachexia. PMID- 11497445 TI - Alterations in systemic and local colonic hemodynamic variables associated with intravenous infusion of ATP-MgCl2 in healthy anesthetized horses. AB - OBJECTIVE: To characterize alterations in systemic and local colonic hemodynamic variables associated with IV infusion of ATP-MgCl2 in healthy anesthetized horses. ANIMALS: 12 adult horses. PROCEDURE: Six horses were given ATP-MgCl2, IV, beginning at a rate of 0.1 mg of ATP/kg of body weight/min with incremental increases until a rate of 1.0 mg/kg/min was achieved. The remaining 6 horses were given an equivalent volume of saline (0.9% NaCl) solution over the same time period. Colonic and systemic hemodynamic variables and colonic plasma nitric oxide (NO) concentrations were determined before, during, and after infusion. RESULTS: Infusion of ATP-MgCl2 caused a rate-dependent decrease in systemic and colonic vascular resistance, principally via its vasodilatory effects. A rate of 0.3 mg of ATP/kg/min caused a significant decrease in systemic and colonic arterial pressure and colonic vascular resistance without a significant corresponding decrease in colonic arterial blood flow. Consistent alterations in NO concentrations of plasma obtained from colonic vasculature were not detected, despite profound vasodilatation of the colonic arterial vasculature. CONCLUSIONS AND CLINICAL RELEVANCE: Results revealed that IV infusion of ATP-MgCl2 may be beneficial in maintaining colonic perfusion in horses with ischemia of the gastrointestinal tract, provided a sufficient pressure gradient exists to maintain blood flow. PMID- 11497446 TI - Effect of mammary secretions on functions of polymorphonuclear leukocytes in pigs. AB - OBJECTIVE: To determine the effects of porcine mammary secretions on polymorphonuclear (PMN) leukocyte function and to relate concentrations of estradiol-17beta and cortisol in mammary secretions to PMN cell function. SAMPLE POPULATION: Mammary secretions from 10 healthy sows and blood PMN leukocytes from 27 healthy sows. PROCEDURE: Mammary secretions were collected within 24 hours after parturition (colostrum) and 12 to 13 days later (milk). Chemoattractant properties were assessed by use of a cell migration assay. Phagocytic capacity of PMN cells in colostrum and milk was assessed by recording chemiluminescence following phagocytosis of Escherichia coli or zymosan. Estradiol-17beta and cortisol concentrations were determined by use of radioimmunoassays. RESULTS: Chemoattractant properties of colostrum and milk were significantly greater than that of zymosan-activated serum. However, chemoattractant properties did not differ significantly between the 2 types of secretions. The capacity of PMN cells in colostrum to phagocytose either zymosan or E. coli was less, compared with cells in milk, and the ability of cells in either type of mammary secretion to phagocytose E. coli was greater than the ability to phagocytose zymosan. Concentrations of estradiol-17beta and cortisol were greater in colostrum, compared with milk. No clear relation was evident between PMN cell activity and hormone concentrations in mammary secretions. CONCLUSIONS AND CLINICAL RELEVANCE: Although chemoattractant properties of colostrum and milk did not differ, the phagocytic capacity of PMN cells in colostrum was significantly less than that of cells in milk. This may predispose sows to coliform mastitis during the early postparturient period. PMID- 11497447 TI - Assessment of lithium dilution cardiac output as a technique for measurement of cardiac output in dogs. AB - OBJECTIVES: To determine agreement of cardiac output measured by use of lithium dilution cardiac output (LiDCO) and thermodilution cardiac output (TDCO) techniques in dogs and to determine agreement of low- and high-dose LiDCO with TDCO. ANIMALS: 10 dogs (7 males, 3 females). PROCEDURE: Cardiac output was measured in anesthetized dogs by use of LiDCO and TDCO techniques. Four rates of cardiac output were induced by occlusion of the caudal vena cava, changes in depth of anesthesia, or administration of dobutamine. Lithium dilution cardiac output was performed, using 2 doses of lithium chloride (low and high dose). Each rate of cardiac output allowed 4 comparisons between LiDCO and TDCO. RESULTS: 160 comparisons were determined of which 68 were excluded. The remaining 92 comparisons had values ranging from 1.10 to 12.80 L/min. Intraclass correlation coefficient (ICC) between low-dose LiDCO and TDCO was 0.9898 and between high dose LiDCO and TDCO was 0.9896. When all LiDCO determinations were pooled, ICC was 0.9894. For determinations of cardiac output < 5.0 L/min, ICC was 0.9730. Mean +/- SD of the differences of TDCO minus LiDCO for all measurements was 0.084+/-0.465 L/min, and mean of TDCO minus LiDCO for cardiac outputs < 5.0 L/min was -0.002+/-0.245 L/min. CONCLUSIONS AND CLINICAL RELEVANCE: The LiDCO technique is a suitable substitute for TDCO to measure cardiac output in dogs. Use of LiDCO eliminates the need for catheterization of a pulmonary artery and could increase use of cardiac output monitoring, which may improve management of cardiovascularly unstable animals. PMID- 11497448 TI - Characteristics and management practices associated with milk production in dairy herds in Ohio enrolled in official Dairy Herd Improvement Association programs. AB - OBJECTIVE: To determine herd characteristics and management practices associated with milk production in dairy herds enrolled in official Dairy Herd Improvement Association (DHIA) programs in Ohio. SAMPLE POPULATION: 186 dairy farms in Ohio. PROCEDURE: All herds in official DHIA programs in 9 counties were invited to participate. Information regarding herd characteristics and management practices was obtained, using a standardized questionnaire. Bulk-tank milk samples were obtained for bacteriologic culture. Official DHIA test-day records were obtained, and associations were identified, using multivariable ANOVA procedures. RESULTS: Of 479 eligible producers, 186 (39%) participated, and consecutive bulk-tank milk samples were available for culture from 172 (36%). Streptococcus agalactiae and Mycoplasma spp were not recovered from bulk-tank milk samples, but Staphylococcus aureus was recovered from 64 (37%) herds. Mean (+/- SD) number of lactating cows in participating herds was 97+/-66, with 123 (66%) herds milking < 100 cows. The RHA was significantly associated with number of cows in milk, estimated percentage of herd detected in estrus, reported annual percentage of heifer calves born alive that died before 8 weeks old, percentage days in milk, use of bovine somatotropin during the preceding 2 years, and sex of the person completing the questionnaire. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, the strongest indicator of milk production was number of cows in milk. However, merely adding cows to a herd should not be considered to guarantee increased milk production, because other management traits could be confounded with increased number of cows in a herd. PMID- 11497449 TI - Changes in protein and nutrient composition of milk throughout lactation in dogs. AB - OBJECTIVE: To evaluate changes in protein and nutrient composition of milk throughout lactation in dogs. SAMPLE POPULATION: Milk samples collected from 10 lactating Beagles. PROCEDURE: Milk samples were collected on days 1, 3, 7, 14, 21, 28, 35, and 42 after parturition and analyzed to determine concentrations of nitrogen, nonprotein nitrogen, casein, whey proteins, amino acids, lipids, lactose, citrate, minerals, and trace elements. Optimum conditions for separating casein from whey proteins and distribution of milk proteins throughout lactation were assessed by use of polyacrylamide gel electrophoresis. RESULTS: Protein concentration was high in samples collected on day 1 (143 g/L), decreased through day 21 (68.4 g/L), and increased thereafter. Concentration of nonprotein nitrogen did not change throughout lactation (5.7 to 9.9% of total nitrogen content). Casein-to-whey ratio was approximately 70:30 and remained constant throughout lactation. Lactose concentration increased from 16.6 g/L on day 1 to 34.0 to 40.2 g/L on days 7 to 42. Lipid concentration ranged from 112.5 to 1372 g/L. Citrate concentration increased from day 1 (4.8 mM) to day 7 (6.6 mM), then gradually decreased until day 42 (3.9 mM). Iron, zinc, copper, and magnesium concentrations decreased during lactation, whereas calcium and phosphorus concentrations increased. Calcium-to-phosphorus ratio remained constant throughout lactation (approx 1.6:1). Energy content of milk ranged from 1,444 to 1,831 kcal/L. CONCLUSIONS AND CLINICAL RELEVANCE: Protein and nutrient composition of milk changes throughout lactation in dogs. These data can provide valuable information for use in establishing nutrient requirements of puppies during the suckling period. PMID- 11497450 TI - Use of monoclonal antibodies to refine flow cytometric differential cell counting of canine bone marrow cells. AB - OBJECTIVES: To evaluate use of monoclonal antibodies to increase accuracy of flow cytometric differential cell counting of canine bone marrow cells. SAMPLE POPULATION: Bone marrow specimens from 15 dogs. PROCEDURES: Specimens were labeled with monoclonal antibodies that detected CD18, major histocompatability antigen class-II (MHC class-II), CD14, and Thy-1. Location of fluorescent and nonfluorescent cells within gates of a template developed for canine bone marrow differential cell counting was determined, the template was revised, and 10 specimens were analyzed by use of the old and revised templates and by labeling cells with anti-MHC class-II and anti-CD14. RESULTS: Data confirmed the presumptive location of marrow subpopulations in scatter plots, permitted detection of lymphocytes and monocytemacrophages, and was used to revise the analysis template used for differential cell counting. When differential cells counts determined by the original and revised templates were compared with results of manual differential cell counts, the revised template had higher correlation coefficients and more similar mean values. Labeling cells with anti MHC class-II and anti-CD14 permitted identification of lymphoid and monocyte macrophages cells in bone marrow specimens. CONCLUSIONS AND CLINICAL RELEVANCE: Use of the revised flow cytometric analysis template combined with anti-CD14 and anti-MHC class-II antibody labeling provides reliable differential cell counts for clinical bone marrow specimens in dogs. These techniques have potential applications to clinical bone marrow examination and preclinical toxicity studies. PMID- 11497451 TI - Detection of shared magnetic antigenic determinants on whole Moraxella bovis pili by use of antisera to cyanogen bromide-cleaved M. bovis pilus protein. AB - OBJECTIVE: To determine the ability of antisera against cyanogen bromide-cleaved pili from 4 strains of Moraxella bovis to react with whole or nondenatured pili. SAMPLE POPULATION: Antisera to 4 strains of M. bovis produced by New Zealand White rabbits. PROCEDURE: Pili from 4 strains of M. bovis were collected and purified. Pilus proteins (pilin) were cleaved, using cyanogen bromide. Whole pilus and cyanogen bromide-cleaved pilin were injected into rabbits. Antisera were serially diluted, reacted with 4 strains of M. bovis, and examined by immunoelectron microscopy and indirect immunofluorescence. RESULTS: Antisera to whole pili aggregated and distorted pili from homologous strains, but pili from heterologous strains were unaffected. Antisera to cleaved pilin fragments resulted in partial aggregation and thickening of homologous and heterologous pili, suggestive of heterospecific antibodies. Attachment of antibodies to pili was detected by indirect immunofluorescence, indicating a strong reaction of antisera to whole pili with homologous pili. Weak cross-reactions were evident with certain heterologous strains. In contrast, antisera to cleaved pilin fragments reacted strongly with pili from homologous and heterologous strains. CONCLUSIONS AND CLINICAL RELEVANCE: We detected shared antigenic determinants on pili from various strains of M. bovis that were not immunogenic in intact pili. These sites were immunogenic after cleavage of pilus protein with cyanogen bromide, and antisera produced to protein fragments reacted with whole pili from heterologous strains of the organism. Vaccines produced from cyanogen bromide treated pili may induce broader immunity against infectious bovine keratoconjuctivitis than that provided by currently available vaccines. PMID- 11497452 TI - Luteal and clinical response following administration of dinoprost tromethamine or cloprostenol at standard intramuscular sites or at the lumbosacral acupuncture point in mares. AB - OBJECTIVE: To determine whether administration of a microdose of prostaglandin at the BAI HUI acupuncture point offers any advantage over IM injections for luteolysis, ovulatory interval, or systemic response in mares. ANIMALS: 17 mature cycling mares, 3 to 20 years of age and weighing 400 to 500 kg. PROCEDURE: Conventional and microdoses of the prostaglandin dinoprost tromethamine (PGF2alpha), the analogue cloprostenol, or sterile water (control) were administered to mares in 7 treatment groups. Treatments were assigned by dose, administration site (semimembranosus, semitendinosus, or lumbosacral region), and treatment type (PGF2alpha, analogue, or sterile water). Mares were observed for ovulatory interval and systemic response to treatment, including heart, and respiratory rates, rectal temperature, and sweat score. Plasma progesterone concentrations were also determined at the time of treatment and at 24-hour intervals for 96 hours following treatment. RESULTS: Ovulatory interval was shortened and progesterone concentrations decreased in prostaglandin-treated mares, compared with control mares, regardless of dose or treatment site. However, no differences in ovulatory interval were observed among prostaglandin treated mares. Mares treated with conventional doses of PGF2alpha had greater systemic responses than mares treated with microdoses of PGF2alpha or sterile water. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of prostaglandins at the BAI HUI acupuncture point does not appear to offer any advantage over administration at standard IM injection sites for induction of luteolysis or to shorten the ovulatory interval. However, administration of a microdose of the analogue cloprostenol was effective at inducing luteolysis and shortening ovulatory interval regardless of administration site. PMID- 11497454 TI - Use of dual energy x-ray absorptiometry for noninvasive body composition measurements in clinically normal dogs. AB - OBJECTIVE: To determine quantitative values for components of body composition in clinically normal dogs of various breeds by use of dual energy x-ray absorptiometry (DEXA) and validate the precision and accuracy of DEXA technology in dogs. ANIMALS: 103 clinically normal sexually intact adult dogs. PROCEDURE: In a cross-sectional study, Beagles, Pembroke Welsh Corgis, Golden Retrievers, Great Danes, Pointers, Rottweilers, and nonpurebred dogs received total body DEXA scans. For the validation portion of the study, the results of DEXA scans of 6 dogs were compared with values obtained by chemical analyses of tissues from euthanatized dogs to determine the accuracy of this modality in dogs. RESULTS: Results (coefficient of variation) of the precision tests ranged from 0.10% for lean tissue to 5.19% for fat tissue, whereas accuracy tests revealed a difference between percentage bone mineral content and ash values. Body composition differed by sex, such as higher lean tissue and bone mineral content in males within some breeds, and among breeds. Regardless of body size or weight, the percentage of body weight that was bone mineral ranged from 3 to 4.0% [corrected]. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this cross-sectional study provide valuable body composition data for clinically normal adult dogs, which may have research and clinical applications. PMID- 11497453 TI - Derivation and characterization of a live attenuated equine influenza vaccine virus. AB - OBJECTIVE: To develop and characterize a cold-adapted live attenuated equine-2 influenza virus effective as an intranasal vaccine. ANIMALS: 8 ponies approximately 18 months of age. PROCEDURES: A wild-type equine-2 virus, A/Equine/Kentucky/1/91 (H3N8), was serially passaged in embryonated chicken eggs at temperatures gradually reduced in a stepwise manner from 34 C to 30 C to 28 C to 26 C. At different passages, infected allantoic fluids were tested for the ability of progeny virus to replicate in Madin-Darby canine kidney (MDCK) cells at 34 C and 39.5 C. Virus clones that replicated at 26 C in eggs and at 34 C in MDCK cells, but not at 39.5 C in MDCK cells, were tested for stability of the cold-adapted, temperature-sensitive (ts), and protein synthesis phenotypes. A stable clone, P821, was evaluated for safety, ability to replicate, and immunogenicity after intranasal administration in ponies. RESULTS: Randomly selected clones from the 49th passage were all ts with plaquing efficiencies of < 10(-6) (ratio of 39.5 C:34 C) and retained this phenotype after 5 serial passages at 34 C in either embryonated eggs or MDCK cells. The clone selected as the vaccine candidate (P821) had the desired degree of attenuation. Administered intranasally to seronegative ponies, the virus caused no adverse reactions or overt signs of clinical disease, replicated in the upper portion of the respiratory tract, and induced a strong serum antibody response. CONCLUSION AND CLINICAL RELEVANCE: A candidate live attenuated influenza vaccine virus was derived by cold-adaptation of a wild-type equine-2 influenza virus, A/Equine/Kentucky/1/91, in embryonated eggs. PMID- 11497455 TI - Use of capillary electrophoresis to quantitate carbamylated hemoglobin concentrations in dogs with renal failure. AB - OBJECTIVE: To evaluate quantification of the amount of carbamylated hemoglobin (CarbHb), using capillary electrophoresis (CE) and a new dynamic capillary coating system to separate hemoglobin derivatives, and to assess the use of CarbHb amounts to evaluate long-term urea exposure and differential diagnoses of azotemia in dogs. ANIMALS: 8 dogs with renal failure, 2 dogs with diabetes mellitus, and 7 control dogs. PROCEDURE: Optimal analytic conditions for separation of CarbHb and other hemoglobin derivatives in blood samples obtained from dogs were determined, using a commercial analysis system developed for the detection of glycohemoglobin Hb A1c (GlycHb) in human blood samples. Relative content of hemoglobin derivatives in blood from 10 dogs with renal failure or endocrine diseases were compared with values for 7 dogs without renal or endocrine diseases. RESULTS: Satisfactory resolution of hemoglobin derivatives was obtained, which permitted identification and quantitation of the amount of CarbHb as a percentage of the total amount of hemoglobin. Normal or increased amounts of GlycHb did not interfere with CarbHb analysis. Dogs with chronic renal failure had considerably higher peak amounts of CarbHb than dogs with acute renal failure, a dog with chronic renal failure that was treated by use of hemodialysis, or dogs without renal disease. CONCLUSIONS AND CLINICAL RELEVANCE: Amounts of CarbHb in blood samples obtained from dogs can be readily quantified by use of capillary electrophoresis. Assessment of the amount of CarbHb can be used to facilitate evaluation of the cause of azotemia in dogs. PMID- 11497456 TI - Evaluation of equine immunoglobulin specific for Rhodococcus equi virulence associated proteins A and C for use in protecting foals against Rhodococcus equi induced pneumonia. AB - OBJECTIVE: To determine whether purified equine immunoglobulin specific for Rhodococcus equi virulence-associated proteins A and C (VapA and VapC) can confer passive protection against R. equi-induced pneumonia in foals. ANIMALS: Twenty eight 3-week-old mixed-breed pony foals. PROCEDURE: 7 foals received IV injections of equine hyperimmune plasma (HIP) against whole-cell R. equi, and 7 received purified equine immunoglobulin specific for VapA and VapC 1 day prior to intrabronchial infection with R. equi strain 103+. Eleven foals were not treated prior to infection, and 3 control foals were neither treated nor infected. Heart rate, respiratory rate, and rectal temperature were recorded twice daily, and serum fibrinogen concentration and WBC count were determined every other day following infection. Foals were euthanatized 14 days following infection, and lung lesions and concentration of R. equi in lungs were assessed. RESULTS: The onset of clinical signs of pneumonia was significantly delayed in the HIP- and immunoglobulin-treated groups, compared with the untreated infected group. Moreover, pulmonary lesions were less severe in the treated groups, and significantly fewer R. equi organisms were cultured from the lungs of treated foals. CONCLUSIONS AND CLINICAL RELEVANCE: Degree of protection against R. equi induced pneumonia provided by purified immunoglobulin specific for VapA and VapC was similar to that provided by commercially available HIP. Results not only suggest that immunoglobulin is the primary component of HIP that confers protection against R. equi-induced pneumonia in foals but also indicate that antibodies against R. equi VapA and VapC are protective. PMID- 11497457 TI - Use of a hyaluronate membrane for jejunal anastomosis in horses. AB - OBJECTIVE: To compare the outcomes of double-layer inverting anastomosis (DIA), single-layer anastomosis (SLA), and single-layer anastomosis combined with a hyaluronate membrane (SLA+HA-membrane) with respect to stomal diameter, adhesion formation, surgery time, and anastomotic healing in horses. ANIMALS: 18 adult horses. PROCEDURE: Midline celiotomy and end-to-end anastomoses were performed. In control horses (n = 6), DIA was performed; in treated horses, SLA was performed (6) or SLA+HA-membrane was performed (6). Horses were euthanatized 21 days after surgery. Abdominal adhesions were evaluated grossly and histologically. Stomal diameters were measured ultrasonographically and compared with adjacent luminal diameters. Anastomotic healing was evaluated histologically for fibrosis and inflammation, tissue alignment, and inversion. Surgery times were recorded for the anastomotic procedure and compared among groups. RESULTS: There were significantly more adhesions in the SLA group, compared with the DIA and SLA+HA-membrane groups. Reduction in stomal diameters in the DIA group was significantly greater than the SLA and SLA+HA-membrane groups. Surgery times for the DIA group were significantly greater than the SLA and SLA+HA-membrane groups. Histologic findings of fibrosis, inflammation, and mucosal healing were similar among groups. There was significant tissue inversion in the DIA group, compared with the 2 treatment groups. Tissue alignment was not different among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Use of a SLA+HA-membrane was an effective small intestinal anastomotic technique. This technique was faster to perform and resulted in a larger stomal diameter, compared with the DIA technique and significantly fewer perianastomotic adhesions, compared with the SLA technique. PMID- 11497458 TI - Use of quantitative analysis of sonographic brightness for detection of early healing of tendon injury in horses. AB - OBJECTIVES: To determine whether quantitative analysis of sonographic brightness could be used to detect healing of an induced injury of the superficial digital flexor tendon in horses and whether rate of healing was influenced by equine recombinant growth hormone. ANIMALS: 8 clinically normal Standardbreds. PROCEDURES: A localized injury was created in the left and right superficial digital flexor tendons of each horse by injection of 2,000 units of collagenase. After injury, 4 horses received equine recombinant growth hormone, a possible promoter of tendon healing. Sonographic images (7.5 MHz) of the flexor tendons and ligaments of the metacarpal region were recorded on videotape prior to injury and weekly for 7 weeks after injury. Images were digitized, and sonographic brightness of tendons and ligaments was calculated. RESULTS: Collagenase-induced injury was sonographically similar to naturally occurring injury. After injury, sonographic brightness of the tendon decreased; after 3 weeks, brightness progressively increased, although by 7 weeks brightness had not returned to preinjury value. Equine recombinant growth hormone had no significant effect on the rate of tendon healing, as evaluated sonographically or at necropsy. CONCLUSIONS AND CLINICAL RELEVANCE: As healing developed, alterations in sonographic brightness of injured tendons coincided with real changes in tendon structure. Quantitative sonographic brightness could be used to accurately monitor healing of equine tendon and ligament injuries and investigate the efficacy of various treatment regimens. PMID- 11497459 TI - Oral vaccination of sexually mature pigs with Brucella abortus vaccine strain RB51. AB - OBJECTIVE: To develop a novel oral vaccine delivery system for swine, using the rough vaccine strain of Brucella abortus. ANIMALS: 56 crossbred pigs from a brucellosis-free facility. PROCEDURE: In 3 separate experiments, pigs were orally vaccinated with doses of 1 x 10(9) to > 1 x 10(11) CFU of strain RB51 vaccine. The vaccine was placed directly on the normal corn ration, placed inside a whole pecan, or mixed with cracked pecans and corn. RESULTS: Oral vaccination of pigs with vaccine strain RB51 resulted in a humoral immune response to strain RB51 and short-term colonization of the regional lymph nodes. CONCLUSIONS AND CLINICAL RELEVANCE: A viscous liquid such as Karo corn syrup in association with pecans that scarify the oral mucosa are necessary when placing the live vaccine directly onto corn or other food rations. Doses of > 1 x 10(11) CFU of RB51 organisms/pig in this mixture ensures 100% colonization of regional lymph nodes via the oral route. This method may allow an efficient and economical means to vaccinate feral swine for brucellosis. PMID- 11497460 TI - Evaluation of plasma clearance of inulin in clinically normal and partially nephrectomized cats. AB - OBJECTIVE: To evaluate accuracy of measuring plasma clearance of inulin as an alternative renal function test for estimation of glomerular filtration rate (GFR) in cats. ANIMALS: 10 cats, first studied with intact kidneys and subsequently studied following partial nephrectomy. PROCEDURE: Clearance studies were performed in 10 clinically normal cats; those same cats then underwent partial nephrectomy, and clearance studies were performed again. Plasma concentration of inulin was determined after administration at 50 mg/kg of body weight to cats while renally intact and 45 mg/kg after the partial nephrectomy. Plasma clearance of inulin (PCin) was determined by dividing the dose by the area under the plasma inulin concentration versus time curve. Results for PCin were compared with values obtained simultaneously for urinary clearance of exogenously administered creatinine (Ccr), a widely accepted method for measurement of GFR in cats with intact kidneys and cats with reduced renal mass. RESULTS: Results of PCin were strongly correlated (r2 = 0.912, P < 0.001) with Ccr. Repeatability of determination of PCin was similar to that of Ccr. Sensitivity and specificity of PCin were superior and equivalent to that of Ccr, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Determination of PCin provides a reliable estimate of GFR in cats and is a promising alternative to determining Ccr in cats. PMID- 11497461 TI - Introduction of the phzH gene of Pseudomonas chlororaphis PCL1391 extends the range of biocontrol ability of phenazine-1-carboxylic acid-producing Pseudomonas spp. strains. AB - Pseudomonas chlororaphis PCL1391 controls tomato foot and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici. Its biocontrol activity is mediated by the production of phenazine-1-carboxamide (PCN). In contrast, the take-all biocontrol strains P. fluorescens 2-79 and P. aureofaciens 30-84, which produce phenazine-1-carboxylic acid (PCA), do not control this disease. To determine the role of the amide group in biocontrol, the PCN biosynthetic genes of strain PCL1391 were identified and characterized. Downstream of phzA through phzG, the novel phenazine biosynthetic gene phzH was identified and shown to be required for the presence of the 1-carboxamide group of PCN because a phzH mutant of strain PCL1391 accumulated PCA. The deduced PhzH protein shows homology with asparagine synthetases that belong to the class II glutamine amidotransferases, indicating that the conversion of PCA to PCN occurs via a transamidase reaction catalyzed by PhzH. Mutation of phzH caused loss of biocontrol activity, showing that the 1-carboxamide group of PCN is crucial for control of tomato foot and root rot. PCN production and biocontrol activity of the mutant were restored by complementing the phzH gene in trans. Moreover, transfer of phzH under control of the tac promoter to the PCA-producing biocontrol strains P. fluorescens 2-79 and P. aureofaciens 30-84 enabled these strains to produce PCN instead of PCA and suppress tomato foot and root rot. Thus, we have shown, for what we believe is the first time, that the introduction of a single gene can efficiently extend the range of the biocontrol ability of bacterial strains. PMID- 11497462 TI - Investigation of myo-inositol catabolism in Rhizobium leguminosarum bv. viciae and its effect on nodulation competitiveness. AB - Three discrete loci required for growth on myo-inositol in Rhizobium leguminosarum bv. viciae have been characterized. Two of these are catabolic loci that code for malonate semialdehyde dehydrogenase (iolA) and malonate semialdehyde dehydrogenase (iolD). IolD is part of a possible operon, iolDEB, although the functions of IolE and IolB are unknown. The third locus, int, codes for an ABC transport system that is highly specific for myo-inositol. LacZ analysis showed that mutation of iolD, which codes for one of the last steps in the catabolic pathway, prevents increased transcription of the entire pathway. It is likely that the pathway is induced by an end product of catabolism rather than myo-inositol itself. Mutants in any of the loci nodulated peas (Pisum sativum) and vetch (Vicia sativa) at the same rate as the wild type. Acetylene reduction rates and plant dry weights also were the same in the mutants and wild type, indicating no defects in nitrogen fixation. When wild-type 3841 was coinoculated onto vetch plants with either catabolic mutant iolD (RU360) or iolA (RU361), however, >95% of the nodules were solely infected with the wild type. The competitive advantage of the wild type was unaffected, even when the mutants were at 100-fold excess. The myo-inositol transport mutant (RU1487), which grows slowly on myo-inositol, was only slightly less competitive than the wild type. The nodulation advantage of the wild type was not the result of superior growth in the rhizosphere. Instead, it appears that the wild type may displace the mutants early on in the infection and nodulation process, suggesting an important role for myo-inositol catabolism. PMID- 11497463 TI - Effects of movement protein mutations on the formation of tubules in plant protoplasts expressing a fusion between the green fluorescent protein and Cauliflower mosaic virus movement protein. AB - Fusions between the green fluorescent protein (GFP) and the Cauliflower mosaic virus (CaMV) movement protein (MP) induce the formation of fluorescent foci and surface tubules in Arabidopsis thaliana leaf mesophyll protoplasts. Tubules elongate coordinately and progressively in an assembly process approximately 6 to 12 h following transfection of protoplasts with GFP-MP constructs. Tubules are not formed in protoplasts transfected by GFP-MP(ER2A), a MP mutation that renders CaMV noninfectious. A small number of short tubules are formed on protoplasts transfected by GFP-MP(N6) and GFP-MP(N13), two second-site revertants of ER2A that partially restore infectivity. Protoplasts cotransfected with cyan fluorescent protein (CFP)-MP(WT) and GFP-MP(ER2A) form tubules containing both MP fusions, indicating that although the GFP-MP(ER2A) cannot induce tubule formation, GFP-MP(ER2A) can coassemble or colocalize with CFP-MP(WT) in tubules. Thus, CaMV MP-induced tubule formation in protoplasts correlates closely with the infectivity of mutation ER2A and its revertants, suggesting that tubule-forming capacity in plant protoplasts reflects a process required for virus infection or movement. PMID- 11497464 TI - Global regulators ExpA (GacA) and KdgR modulate extracellular enzyme gene expression through the RsmA-rsmB system in Erwinia carotovora subsp. carotovora. AB - The production of the main virulence determinants, the extracellular plant cell wall-degrading enzymes, and hence virulence of Erwinia carotovora subsp. carotovora is controlled by a complex regulatory network. One of the global regulators, the response regulator ExpA, a GacA homolog, is required for transcriptional activation of the extracellular enzyme genes of this soft-rot pathogen. To elucidate the mechanism of ExpA control as well as interactions with other regulatory systems, we isolated second-site transposon mutants that would suppress the enzyme-negative phenotype of an expA (gacA) mutant. Inactivation of kdgR resulted in partial restoration of extracellular enzyme production and virulence to the expA mutant, suggesting an interaction between the two regulatory pathways. This interaction was mediated by the RsmA-rsmB system. Northern analysis was used to show that the regulatory rsmB RNA was under positive control of ExpA. Conversely, the expression of rsmA encoding a global repressor was under negative control of ExpA and positive control of KdgR. This study indicates a central role for the RsmA-rsmB regulatory system during pathogenesis, integrating signals from the ExpA (GacA) and KdgR global regulators of extracellular enzyme production in E. carotovora subsp. carotovora. PMID- 11497465 TI - Dark green islands in plant virus infection are the result of posttranscriptional gene silencing. AB - Dark green islands (DGIs) are a common symptom of plants systemically infected with a mosaic virus. DGIs are clusters of green leaf cells that are free of virus but surrounded by yellow, virus-infected tissue. We report here on two lines of evidence showing that DGIs are caused by posttranscriptional gene silencing (PTGS). First, transcripts of a transgene derived from the coat protein of Tamarillo mosaic potyvirus (TaMV) were reduced in DGIs relative to adjacent yellow tissues when the plants were infected with TaMV. Second, nontransgenic plants coinfected with TaMV and a heterologous virus vector carrying TaMV sequences showed reduced titers of the vector in DGIs compared with surrounding tissues. DGIs also were compared with recovered tissue at the top of transgenic plants because recovery has been shown previously to involve PTGS. Cytological analysis of the cells at the junction between recovered and infected tissue was undertaken. The interface between recovered and infected cells had very similar features to that surrounding DGIs. We conclude that DGIs and recovery are related phenomena, differing in their ability to amplify or transport the silencing signal. PMID- 11497466 TI - Maize rhm1 resistance to Bipolaris maydis is associated with few differences in pathogenesis-related proteins and global mRNA profiles. AB - The maize rhm1 mutant resists Bipolaris maydis, the causal agent of Southern corn leaf blight, by producing small necrotic lesions surrounded by chlorotic haloes. The rhm1 and wild-type lesions contain viable fungus in equal frequency, but fungal sporulation was markedly inhibited on rhm1. The levels of the pathogenesis related (PR) proteins chitinase, PR1, and peroxidase differ little between rhm1 and wild type, with or without B. maydis inoculation. The global mRNA profiles surveyed revealed hundreds of cDNA fragments that were twofold or more induced or suppressed in rhm1 and wild-type plants following B. maydis inoculation. Nonetheless, between rhm1 and wild type, only 0.4 to 0.7% of the cDNA fragments were expressed differentially by twofold or more. Among the up-regulated genes in rhm1 was beta-glucosidase glu1, which prompted a test of whether rhm1 resistance depends upon the antimicrobial compound 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3 one or other hydroxamic acids whose glucosyl conjugates are preferred substrates for the Glu1 enzyme. Double mutants of rhm1 and bx1, a hydroxamic acid-deficient mutant, indicate that rhm1 resistance is hydroxamic acid independent. The rhm1 resistance presently appears to operate via a mechanism unlike those of previously described resistance genes. PMID- 11497467 TI - Inhibition of growth of Aspergillus flavus and fungal alpha-amylases by a lectin like protein from Lablab purpureus. AB - Aspergillus flavus is a fungal pathogen of maize causing an important ear rot disease when plants are exposed to drought and heat stress. Associated with the disease is the production of aflatoxins, which are a series of structurally related mycotoxins known to be carcinogenic. Previous research has suggested that the alpha-amylase of A. flavus promotes aflatoxin production in the endosperm of infected maize kernels. We report here the isolation and characterization of a 36 kDa alpha-amylase inhibitor from Lablab purpureus (AILP). AILP inhibited the alpha-amylases from several fungi but had little effect on those from animal and plant sources. The protein inhibited conidial germination and hyphal growth of A. flavus. The amino acid sequence indicated that AILP is similar to lectin members of a lectin-arcelin-alpha-amylase inhibitor family described in common bean and shown to be a component of plant resistance to insect pests. AILP also agglutinated papain-treated red blood cells from human and rabbit. These data indicate that AILP represents a novel variant in the lectin-arcelin-alpha-amylase inhibitor family of proteins having lectin-like and alpha-amylase inhibitory activity. PMID- 11497468 TI - Type III secretion contributes to the pathogenesis of the soft-rot pathogen Erwinia carotovora: partial characterization of the hrp gene cluster. AB - The virulence of soft-rot Erwinia species is dependent mainly upon secreted enzymes such as pectinases, pectin lyases, and proteases that cause maceration of plant tissue. Some soft-rot Erwinia spp. also harbor genes homologous to the hypersensitive reaction and pathogenesis (hrp) gene cluster, encoding components of the type III secretion system. The hrp genes are essential virulence determinants for numerous nonmacerating gram-negative plant pathogens but their role in the virulence of soft-rot Erwinia spp. is not clear. We isolated and characterized 11 hrp genes of Erwinia carotovora subsp. carotovora. Three putative sigmaL-dependent Hrp box promoter sequences were found. The genes were expressed when the bacteria were grown in Hrp-inducing medium. The operon structure of the hrp genes was determined by mRNA hybridization, and the results were in accordance with the location of the Hrp boxes. An E. carotovora strain with mutated hrcC, an essential hrp gene, was constructed. The hrcC- strain was able to multiply and cause disease in Arabidopsis, but the population kinetics were altered so that growth was delayed during the early stages of infection. PMID- 11497469 TI - Phenazine-1-carboxamide production in the biocontrol strain Pseudomonas chlororaphis PCL1391 is regulated by multiple factors secreted into the growth medium. AB - Pseudomonas chlororaphis PCL1391 controls tomato foot and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici. The production of phenazine-1 carboxamide (PCN) is crucial for this biocontrol activity. In vitro production of PCN is observed only at high-population densities, suggesting that production is under the regulation of quorum sensing. The main autoinducer molecule produced by PCL1391 was identified structurally as N-hexanoyl-L-homoserine lactone (C6-HSL). The two other autoinducers that were produced comigrate with N-butanoyl-L homoserine lactone (C4-HSL) and N-octanoyl-L-homoserine lactone (C8-HSL). Two PCL1391 mutants lacking production of PCN were defective in the genes phzI and phzR, respectively, the nucleotide sequences of which were determined completely. Production of PCN by the phzI mutant could be complemented by the addition of exogenous synthetic C6-HSL, but not by C4-HSL, C8-HSL, or any other HSL tested. Expression analyses of Tn5luxAB reporter strains of phzI, phzR, and the phz biosynthetic operon clearly showed that phzI expression and PCN production is regulated by C6-HSL in a population density-dependent manner. The introduction of multiple copies of the regulatory genes phzI and phzR on various plasmids resulted in an increase of the production of HSLs, expression of the PCN biosynthetic operon, and consequently, PCN production, up to a sixfold increase in a copy-dependent manner. Surprisingly, our expression studies show that an additional, yet unidentified factor(s), which are neither PCN nor C4-HSL or C8 HSL, secreted into the growth medium of the overnight cultures, is involved in the positive regulation of phzI, and is able to induce PCN biosynthesis at low cell densities in a growing culture, resulting in an increase of PCN production. PMID- 11497470 TI - Cultivar-dependent expression of a maize lipoxygenase responsive to seed infesting fungi. AB - Maize kernels are highly susceptible to Aspergillus spp. infection and aflatoxin (AF) contamination. Fatty acid signaling molecules appear to mediate the plant fungal interaction by affecting the growth, development, and AF production of the fungus. In particular, fatty acid derivatives of the plant lipoxygenase (LOX) pathway are implicated in the Aspergillus spp.-seed interaction. The 9(S) hydroperoxide derivative of linoleic acid promotes transcription of AF genes, whereas the 13(S)-hydroperoxide derivative decreases AF gene expression and production; both are sporulation factors. Our goal was to identify LOX genes responsive to Aspergillus spp. colonization and determine their specificities, 9(S)- or 13(S)-. Screening maize LOX expressed sequence tags (ESTs) identified one clone, cssap 92, which is highly expressed in Aspergillus spp.-infected seed susceptible to AF contamination and repressed in lines with resistance to AF contamination. The accumulation of cssap 92 transcript was similar during Fusarium spp. infection. The cDNA clone has 94% identity to the previously described L2 LOX gene from maize. Product-specificity analysis of the CSSAP 92 protein shows that it preferentially adds oxygen to carbon 9 of linoleic acid. Because 9(S)-hydroperoxy linoleic acid has been implicated as an aflatoxin signaling molecule, it is possible that cssap 92 could be used as a biomarker that is indicative of AF resistance in maize lines. PMID- 11497471 TI - Colletotrichum gloeosporioides pelB is an important virulence factor in avocado fruit-fungus interaction. AB - Colletotrichum gloeosporioides is an important pathogen of tropical and subtropical fruits. The C. gloeosporioides pelB gene was disrupted in the fungus via homologous recombination. Three independent isolates, GD-14, GD-23, and GD 29, did not produce or secrete pectate lyase B (PLB) and exhibited 25% lower pectate lyase (PL) and pectin lyase (PNL) activities and 15% higher polygalacturonase (PG) activity than the wild type. The PLB mutants exhibited no growth reduction on glucose, Na polypectate, or pectin as the sole carbon source at pH 3.8 or 6.0, except for a 15% reduction on pectin at pH 6.0. When pelB mutants were inoculated onto avocado fruits, however, a 36 to 45% reduction in estimated decay diameter was observed compared with the two controls, the wild type and undisrupted transformed isolate. In addition, these pelB mutants induced a significantly higher host phenylalanine ammonia lyase activity as well as the antifungal diene, which is indicative of higher host resistance. These results suggest that PLB is an important factor in the attack of C. gloeosporioides on avocado fruit, probably as a result of its virulence factor and role in the induction of host defense mechanisms. PMID- 11497472 TI - Ancient diversification of the Pto kinase family preceded speciation in Solanum. AB - Recent phylogenetic analyses of the nucleotide binding sites (NBS)-leucine-rich repeats (LRR) class of plant disease resistance (R) genes suggest that these genes are ancient and coexist next to susceptibility alleles at resistance loci. Another class of R genes encodes serine-threonine protein kinases related to Pto that were originally identified from wild relatives of tomato. In this study, we exploit the highly diverse genus Solanum to identify Pto-like sequences and test various evolutionary scenarios for Pto-like genes. Polymerase chain reaction amplifications with the use of primers that were developed on the basis of conserved and variable regions of Pto revealed an extensive Pto gene family and yielded 32 intact Pto-like sequences from six Solanum species. Furthermore, Pto like transcripts were detected in the leaf tissue of all tested plants. The kinase consensus and autophosphorylation sites were highly conserved, in contrast to the kinase activation domain, which is involved in ligand recognition in Pto. Phylogenetic analyses distinguished nine classes of Pto-like genes and revealed that orthologs were more similar than paralogs, suggesting that the Pto gene family evolved through a series of ancient gene duplication events prior to speciation in Solanum. Thus, like the NBS-LRR class, the kinase class of R genes is highly diverse and ancient. PMID- 11497473 TI - Role of repeated fine-needle aspiration of thyroid nodules with benign cytologic features. AB - OBJECTIVE: To determine the role of repeated fine-needle aspiration (FNA) biopsy in the evaluation of thyroid nodules initially classified as benign. METHODS: We retrospectively reviewed data on 235 patients with clinically palpable thyroid nodules who underwent a repeated FNA biopsy after an initially benign diagnosis. All the nodules were evaluated and biopsies were obtained by the same endocrinologist. Cytodiagnoses were divided into four major categories: inadequate, benign, suspicious, or malignant. RESULTS: The study cohort consisted of 211 female and 24 male patients with a mean age of 47.1 years. The repeated FNA cytodiagnoses were as follows: 204 (86.8%) remained benign and 19 (8.1%), 11 (4.7%), and 1 (0.4%) became inadequate, suspicious, and malignant, respectively. All patients with benign or inadequate cytologic results on repeated FNA who underwent thyroid surgical resection had benign histologic findings (N = 23). The mean follow-up period between the initial and the last benign FNA cytodiagnosis in the 186 patients without surgical intervention was 1,078 days (2.95 years). Nine of 11 patients with suspicious cytologic results underwent a thyroid surgical procedure, which revealed a benign lesion in 7 and malignant disease in 2 (18%). The other two patients with suspicious cytologic findings had a 13-year clinical follow-up without any clinical evidence of a thyroid malignant lesion. The only patient with malignant cytologic findings on repeated FNA (a 76-year-old woman) refused surgical treatment and was lost to follow-up. CONCLUSION: Although the yield of finding a malignant lesion on repeated FNA biopsy in the follow-up of a presumably benign thyroid nodule may be low, rebiopsy reduces the rate of false-negative diagnosis from an average of 5.2% to <1.3%. PMID- 11497474 TI - Neck and whole-body scanning with 5-mCi dose of (123)I as diagnostic tracer in patients with well-differentiated thyroid cancer. AB - OBJECTIVE: To determine whether a 5-mCi dose of 123I can be used as an effective radiotracer for assessing the presence of remnant thyroid tissue and for searching for metastatic lesions in patients with well-differentiated thyroid cancer as well as to attempt to ascertain whether a scan performed only at 4 hours is sufficient for accurate diagnosis and might replace the conventional protocol of scanning at both 4 hours and 24 hours. METHODS: We prospectively studied 27 patients who had undergone near-total thyroidectomy and had a documented diagnosis of well-differentiated thyroid carcinoma. Patients underwent scanning after receiving a 5-mCi dose of 123I, at a time when they had discontinued thyroid replacement therapy and had a thyrotropin level in excess of 30 mIU/mL. Whole-body images at 4 hours and 24 hours were obtained and were compared with posttherapy scans obtained 5 to 7 days after administration of 131I. Scans were interpreted by two board-certified nuclear medicine physicians. RESULTS: Of the 27 patients, 2 (7.4%) showed discordance between the 123I scan performed at 24 hours and the posttherapy 131I scan. When 4-hour images after administration of 123I were compared with the posttherapy 131I scans, a discordance rate of 14.8% (4 of 27 patients) was noted. In addition, two of these four patients showed lesions on the 24-hour images that were not seen on the 4 hour images (one with new lung metastatic involvement and the other with a local recurrence in the lower neck area). The prognosis and treatment of these two patients were substantially changed by the result of the 24-hour images. CONCLUSION: On comparison of scans obtained after administration of a 5-mCi dose of 123I with those obtained after 131I therapy, we conclude that 5 mCi of 123I produces images that have excellent quality and resolution and also compare favorably with those obtained after 131I therapy. Furthermore, a decrease in the dose of 123I from 10 mCi to 5 mCi lowered the cost of the study without compromising the diagnostic accuracy or image quality. Finally, use of 24-hour images will occasionally disclose additional areas of radioiodine uptake not detected on the 4-hour scans and is therefore recommended. PMID- 11497475 TI - Meaning of illness and health outcomes in type 1 diabetes. AB - OBJECTIVE: To determine whether a relationship exists between the meaning attributed to type 1 diabetes and mental and physical health outcomes. METHODS: The study sample consisted of 49 adults with type 1 diabetes. Each participant voluntarily agreed to complete the Meaning of Illness Questionnaire (MIQ), the Short Form-36 Health Survey (SF-36), and the Diabetes Health Survey. Multivariant analysis of covariance was used to determine the relationships among the MIQ and SF-36, a history of depression, hemoglobin A1c, and other demographic variables. RESULTS: The meaning attributed to illness predicted health outcomes. Specifically, the impact of illness (MIQ subscale 1; P = 0.030) predicted SF-36 physical functioning, bodily pain, and general mental health; the degree of stress (MIQ subscale 3; P = 0.008) predicted SF-36 general physical health, vitality, and general mental health. Conversely, a history of depression (P = 0.014) and high hemoglobin A1c (P = 0.039) predicted a more negative meaning attributed to illness. CONCLUSION: The meaning attributed to illness varies with physical and mental health. In addition, physical and mental health outcomes and hemoglobin A1c, a measure of mean blood glucose, affect the meaning attributed to diabetes. The findings in this study support a bidirectional relationship between the meaning attributed to illness and health outcomes in patients with type 1 diabetes. PMID- 11497477 TI - Interplay of pregnancy, lactation, and hyperthyroidism leading to severe osteoporosis in a young woman. AB - OBJECTIVE: To describe the case of a young woman who had severe osteoporosis due to the compounding effects of pregnancy, lactation, and hyperthyroidism and who had a presumed metastatic lesion in the lumbar spine. METHODS: We present the clinical, pathologic, radiologic, and laboratory findings and describe the clinical course of our patient. RESULTS: A 31-year-old Arabic woman was referred to the M. D. Anderson Cancer Center because of a lytic lesion in her lumbar spine, presumed to be metastatic deposits. She had a history of two consecutive pregnancies and intermittently treated hyperthyroidism. Our initial evaluation revealed that the patient had clinical and biochemical thyrotoxicosis, and we treated her with thionamides, corticosteroids, and radioiodine ablation. Radiologic studies disclosed a complex renal cyst that had increased uptake on a bone scan, which was highly suggestive of a primary malignant lesion. Ultimately, however, it proved benign on pathologic analysis after a left nephrectomy. Bone mineral density measurements identified severe osteoporosis (T-scores: lumbar spine, -3.3; right hip, -2.2; and left hip, -2.0), which had led to vertebral collapse and was misinterpreted as malignant metastatic disease. The bone mineral densities improved (+5 to +11% at the various sites) within 4 months after definitive treatment and cure of the hyperthyroidism. CONCLUSION: The effect of pregnancies and prolonged lactation, in the milieu of other risk factors for bone depletion such as hyperthyroidism, may cause severe osteoporosis in a young patient. The resulting osteoporosis may manifest as a lesion suggestive of malignant metastatic involvement. PMID- 11497476 TI - One-hour meal tolerance test to assess withdrawal of insulin therapy in overweight patients with type 2 diabetes. AB - OBJECTIVE: To evaluate the clinical usefulness of a 1-hour meal tolerance test to determine the feasibility and safety of discontinuation of insulin therapy in overweight, insulin-treated patients with type 2 diabetes mellitus. METHODS: Overweight patients who presented with metabolic decompensation at initial diagnosis of type 2 diabetes were hospitalized and treated with insulin. A 1-hour meal tolerance test was performed in an outpatient setting when patients had maintained a stable metabolic status with decreasing daily insulin requirement and the possibility of withdrawal of insulin therapy was considered. After an overnight fast, venous blood samples were collected before and 1 hour after consumption of an oral liquid meal (240 mL, 240 kcal, approximately 30 g of sucrose) to measure glucose and C-peptide levels. If the postmeal glucose level was < or = 10 mmol/L, the insulin therapy was discontinued. Patients underwent follow-up to validate the clinical decision. RESULTS: Twenty-four overweight adult Mexican American patients (8 of whom had ketoacidosis) were treated with insulin at initial diagnosis of type 2 diabetes. The mean insulin dosages were reduced progressively from 71 +/- 8 U/day to 31 +/- 3 U/day during a period of 4 +/- 0.5 months. At that stage, the mean fasting and 1-hour postmeal blood glucose levels were 5.0 +/- 0.2 mmol/L and 8.4 +/- 0.4 mmol/L, respectively. Meal-induced plasma C-peptide levels showed 1.9- to 4-fold increases, confirming substantial endogenous insulin secretion. The C-peptide responses were similar in the two subgroups of patients with or without initial ketoacidosis. On the basis of postmeal blood glucose levels, the insulin therapy was discontinued in all cases. After 3 months of follow-up, all patients were in good metabolic control. Fourteen patients underwent follow-up for >12 months; 10 of them were maintained without antidiabetic medications for 21.4 +/- 2.8 months. Two patients were treated with sulfonylurea drugs after 4 months, and another two required insulin therapy by the 4th and 18th months, respectively. CONCLUSION: A 1-hour postmeal blood glucose level is helpful for deciding the withdrawal of insulin therapy in overweight, insulin-treated patients with type 2 diabetes. PMID- 11497478 TI - Occult Leydig cell tumor in a patient with gynecomastia. AB - OBJECTIVE: To report a case of a clinically occult testicular tumor causing gynecomastia and to alert physicians to the importance of use of testicular ultrasonography in patients with progressive gynecomastia despite normal findings on testicular examination. METHODS: We present a detailed case, including results of clinical, laboratory, and radiologic assessment, of a man with hyperprolactinemia and gynecomastia. RESULTS: A 36-year-old man with progressive gynecomastia was referred to our clinic because of an increased serum prolactin level. Subsequent clinical investigation revealed no evidence of hypogonadism and several possible causes of the gynecomastia. Because of the patient's age and progressive symptoms, testicular ultrasonography was performed despite normal findings on testicular examination. This ultrasound study showed a right testicular mass, which proved to be a Leydig cell tumor. The patient was referred for definitive therapy with orchiectomy. Follow-up studies showed resolution of the gynecomastia and substantial decreases in prolactin and estradiol levels. CONCLUSION: Although gynecomastia is a relatively common disorder with a benign cause in most cases, physicians should be aware that normal findings on testicular examination do not completely rule out the possibility of a testicular tumor as the cause. Because of the potentially high morbidity of testicular tumors and their known association with gynecomastia, early performance of testicular ultrasonography in a patient with gynecomastia of unknown cause is advised. PMID- 11497479 TI - Ectopic primary hyperparathyroidism. AB - OBJECTIVE: To review an unusual case of ectopic primary hyperparathyroidism. METHODS: We present a case report, including longitudinal results of laboratory studies, and comment on the difficulties with standard imaging techniques, interpretation of parathyroid pathologic findings, and use of invasive localization techniques. RESULTS: A 74-year-old woman underwent repeated assessments because of hypercalcemia. When standard parathyroid 99mTc sestamibi imaging was negative for adenoma and initial surgical exploration of the neck and removal of 3' parathyroid glands were unsuccessful in correcting the hyperparathyroidism, the ectopic adenoma was ultimately localized on an extended field 99mTc sestamibi scan in the region of the right dome of the diaphragm. The localization was confirmed by selective venous sampling and angiography. The ectopic parathyroid adenoma was embolized, and 1 month later, the serum calcium level was normal. CONCLUSION: Ectopic lesions should be considered when standard imaging techniques are nonrevealing and standard neck exploration fails to disclose a parathyroid adenoma in a patient with persistent hypercalcemia. Selective venous sampling and angiography can assist in localization of ectopic parathyroid adenomas. PMID- 11497480 TI - Myasthenia gravis in conjunction with Graves' disease: a diagnostic challenge. AB - OBJECTIVE: To describe an association between Graves' disease and myasthenia gravis and discuss the clinical features and laboratory tests that may help distinguish these two diseases. METHODS: The clinical, laboratory, and electrophysiologic findings in a patient with Graves' disease and myasthenia gravis are presented. RESULTS: A 28-year-old African American man was admitted to the University of Louisville Hospital with generalized muscle weakness, exophthalmos, diplopia, weight loss, and mild dysphagia. The diagnosis of Graves' disease with ophthalmologic involvement was suspected clinically and confirmed by an undetectable thyrotropin level (<0.03 mIU/mL), high total thyroxine (20.5 mg/dL), and increased homogeneous 123I thyroid uptake. Because of the generalized muscle weakness and mild dysphagia, assessment was done by a neurology team, and severe thyrotoxic myopathy was diagnosed. He was treated with 131I and b adrenergic blocking agents and scheduled for follow-up as an outpatient. Two weeks later, the patient presented in acute respiratory failure. The neurology team was reconsulted because of suspected myasthenic crisis. Anti-acetylcholine receptor antibodies were undetectable in the serum, and computed tomography of the chest showed no thymic enlargement. Repetitive nerve stimulation testing, however, showed findings consistent with an abnormality of the neuromuscular junction. The patient responded dramatically to an anticholinesterase agent and corticosteroids. CONCLUSION: The overlapping clinical features may cause diagnostic confusion when myasthenia gravis and Graves' disease coexist, and numerous tests may be needed to distinguish these two conditions, which have differing treatments and prognoses. PMID- 11497481 TI - Use of insulin-sensitizing agents in patients with polycystic ovary syndrome. AB - OBJECTIVE: To review the subject of polycystic ovary syndrome and the therapeutic use of insulin-sensitizing agents in patients with this endocrinopathy. METHODS: We present background information on this disorder and summarize the pertinent published literature. RESULTS: Polycystic ovary syndrome affects approximately 7.5% of reproductive-age women in the United States. Although specific diagnostic criteria for this condition have not been established, the presence of three major factors-chronic anovulation, hyperandrogenemia, and clinical signs of hyperandrogenism-has been proposed as essential for consideration of the diagnosis. A high ratio of serum luteinizing hormone to follicle-stimulating hormone is found in 60 to 75% of women with this syndrome. Treatment with metformin may yield heterogeneous responses in differing populations with polycystic ovary syndrome, but most studies have shown evidence of restoration of ovulatory cycling. In addition, weight loss and decreases in free and total testosterone levels have been reported. Troglitazone therapy proved somewhat less efficacious than metformin for restoring menstrual cycles and similar to metformin in producing hormonal responses. Because troglitazone is no longer available for clinical use, studies will need to be extended to other thiazolidinediones. Patients treated with another insulin sensitizer, D-chiro inositol, have demonstrated improved insulin sensitivity, ovulatory rates, and biochemical findings. CONCLUSION: Current evidence suggests that the use of insulin-sensitizing agents in patients with polycystic ovary syndrome not only improves their sensitivity to the effects of insulin on glucose and lipid metabolism but also ameliorates clinical and biochemical manifestations of hyperandrogenism and increases rates of ovulation. Multicenter studies with larger numbers of patients are needed. PMID- 11497482 TI - From artisan to architect: the specialist and systems of provision of diabetes care in 2001. AB - OBJECTIVE: To discuss the future role of the diabetes specialist and the endocrinologist in the care of patients with diabetes. METHODS: We reviewed the literature on health care systems, integrated approaches to provision of health care, health care in the primary-care setting for patients with chronic illness, and population-based health care for patients with diabetes. RESULTS: Specialists who care for patients with diabetes will assume a more significant role as caretakers of teams of health care professionals who are responsible for the primary care of patients with diabetes. The integration of specialist care at the primary-care level may lead to improved outcomes at the primary-care level, increased access to specialty care, and decreased costs for the health care system. Alternative systems, such as regulatory barriers to specialist care, are less likely to accomplish these outcomes. CONCLUSION: In the near future, specialists will be expected to care for those who provide primary care for patients with diabetes. Endocrinologists and endocrinology training programs should recognize this emerging role and prepare for it. PMID- 11497483 TI - Primary hyperparathyroidism: new options for surgical management. PMID- 11497484 TI - Rebiopsy of benign thyroid nodules: optimal approach to maximize safety and minimize regret. PMID- 11497485 TI - Radioiodine and thyroid cancer: some questions, controversies, and considerations. PMID- 11497486 TI - Surgical treatment of primary hyperparathyroidism. PMID- 11497487 TI - Conversion from troglitazone to rosiglitazone. PMID- 11497488 TI - Femoral preparation in cemented total hip arthroplasty: reaming or broaching? AB - Surgical techniques continue to be refined to improve the results of primary cemented total hip arthroplasty. Although there has been much research in the areas of cementation and implant design, little work has specifically addressed how bone preparation can be optimized on the femoral side. On the basis of available scientific data, it appears that the broach-only system has several potential advantages over the traditional ream-and-broach technique. Broaching is usually faster, leaves behind more bone stock, and may improve both microinterlock and macrointerlock. Additionally, the excess bone resulting from broaching without reaming does not seem to compromise fixation at the bone-cement interface. Such differences may become even more important as the indications for cemented hip arthroplasty broaden to include increasingly younger and more active patients, because revision in these individuals is likely. In most cases, reaming is probably counterproductive, although it may be advantageous when used to open the femoral canal, to prevent varus stem orientation, and to manage sclerosis or deformity of bone due to a preexisting hip disorder or the presence of internal fixation devices. Regardless of which method is chosen, good bone surface cleansing and cement penetration remain paramount. More studies comparing reamed and nonreamed preparation are necessary to resolve this controversial issue definitively. PMID- 11497489 TI - Suprascapular neuropathy. AB - Suprascapular neuropathy is an uncommon cause of shoulder pain and weakness and therefore may be overlooked as an etiologic factor. The suprascapular nerve is vulnerable to compression at the suprascapular notch as well as at the spinoglenoid notch. Other causes of suprascapular neuropathy include traction injury at the level of the transverse scapular ligament or the spinoglenoid ligament and direct trauma to the nerve. Sports involving overhead motion, such as tennis, swimming, and weight lifting, may result in traction injury to the suprascapular nerve, leading to dysfunction. The diagnosis of suprascapular neuropathy is based on clinical findings and abnormal electrodiagnostic test results, after the exclusion of other causes of shoulder pain and weakness. Magnetic resonance imaging may provide an anatomic demonstration of nerve entrapment and muscle atrophy. With this modality, ganglion cysts are recognized with increasing frequency as a source of external compression of the suprascapular nerve. Without evidence of a discrete lesion compressing the nerve, nonoperative treatment should include physical therapy and avoidance of precipitating activities. When nonoperative treatment fails to alleviate symptoms or when a discrete lesion such as a ganglion cyst is present, surgical decompression is warranted. Decompression gives reliable pain relief, but recovery of shoulder function and restoration of atrophied muscle tissue may be incomplete. PMID- 11497490 TI - Surgical management of cervical radiculopathy. AB - Cervical radiculopathy presents as pain in a dermatomal distribution. Despite conservative nonoperative therapy, a large subset of patients will require surgical intervention. Indications for surgery include recalcitrant radiculopathy despite nonoperative treatment for more than 6 weeks and progressive motor deficit or disabling motor deficit (deltoid palsy, wrist drop) prior to 6 weeks. Anterior and posterior approaches have both yielded successful results in appropriately selected patients. Anterior cervical diskectomy and fusion is the generally preferred treatment for radiculopathy when there is a significant component of axial neck pain, when the disease is centrally located, or when there is any degree of segmental kyphosis. Posterior laminoforaminotomy is an acceptable choice for lateral soft disk herniations with predominant arm pain and for caudal lesions in large, short-necked individuals. PMID- 11497491 TI - Peptide insertions in domain 4 of hbeta(c), the shared signalling receptor subunit for GM-CSF, IL3 and IL5, induce ligand-independent activation. AB - A mutant form of the common beta-subunit of the GM-CSF, interleukin-3 (IL3) and IL5 receptors is activated by a 37 residue duplicated segment which includes the WSXWS motif and an adjacent, highly conserved, aliphatic/basic element. Haemopoietic expression of this mutant, hbeta(c)FIDelta, in mice leads to myeloproliferative disease. To examine the mechanism of activation of this mutant we targetted the two conserved motifs in each repeat for mutagenesis. Here we show that this mutant exhibits constitutive activity in BaF-B03 cells in the presence of mouse or human GM-CSF receptor alpha-subunit (GMRalpha) and this activity is disrupted by mutations of the conserved motifs in the first repeat. In the presence of these mutations the receptor reverts to an alternative conformation which retains responsiveness to human IL3 in a CTLL cell line co expressing the human IL3 receptor alpha-subunit (hIL3Ralpha). Remarkably, the activated conformation is maintained in the presence of substitutions, deletions or replacement of the second repeat. This suggests that activation occurs due to insertion of extra sequence after the WSXWS motif and is not dependent on the length or specific sequence of the insertion. Thus hbeta(c) displays an ability to fold into functional receptor conformations given insertion of up to 37 residues in the membrane-proximal region. Constitutive activation most likely results from a specific conformational change which alters a dormant, inactive receptor complex, permitting functional association with GMRalpha and ligand independent mitogenic signalling. PMID- 11497492 TI - Functional analysis of linker-scan mutants spanning the -376, -308, -244, and 238 polymorphic sites of the TNF-alpha promoter. AB - Tumor necrosis factor alpha (TNF-alpha) promoter polymorphisms have been linked to a large number of diseases but studies examining the possible direct functional effects of these polymorphisms have been contradictory. Previous studies compared TNF-alpha promoter constructs containing single nucleotide changes. We have now made a series of mutant constructs in which regions of the TNF-alpha promoter containing suspected functional single nucleotide polymorphisms, including -376, -308, -244 and -238, were replaced by a 10 bp linker scan sequence. These constructs were transiently transfected into the T cell line Jurkat, the B cell line Raji, and the monocytic cell line U937, and tested for basal and induced transcriptional activity. Mutant constructs covering both the -308 and -376 polymorphisms showed no significant differences in either basal or induced transcriptional activity. Constructs covering the -244/-238 region showed a small increase in basal activity in the U937 cell line. These results indicate (i) that the -308 and -376 regions are of no functional relevance for TNF-alpha promoter transcription, and (ii) that the -244/-238 region does not influence transcription in some cell lines but may have some role in transcription in others. PMID- 11497493 TI - 17-beta-estradiol suppresses IL-2 and IL-2 receptor. AB - Interleukin-2 (IL-2) plays an important role in adaptive immune responses. These responses differ between females and males and may be due to the sex steroid estrogen. In this investigation we show that estrogen suppresses IL-2 production from activated peripheral blood T cells and CD4+ T cell lines at the transcriptional level. Suppression of IL-2 occurred at short term, high 17-beta estradiol concentrations as well as longer term lower 17-beta-estradiol concentrations. In CD4+ Jurkat T cells, suppression of IL-2 was associated with decreased nuclear binding of two important IL-2 promoter transcription factors: NFkappaB and AP-1. The decreased nuclear binding of NFkappaB occurred in the setting of estrogen-induced increases in IkappaBalpha protein levels, an important inhibitor of NFkappaB nuclear translocation. 17-beta-Estradiol was also shown to inhibit IL-2 receptor (IL-2R) expression in activated peripheral blood T cells. Estrogen-induced suppression of IL-2 and its receptor may have many ramifications for our understanding of immune and autoimmune sexual dichotomies, immune responses during pregnancy, and potential therapeutic intervention with hormone agonists and antagonists. PMID- 11497494 TI - A novel IL-18BP ELISA shows elevated serum IL-18BP in sepsis and extensive decrease of free IL-18. AB - IL-18 binding protein (IL-18BP) is a circulating antagonist of the proinflammatory Th1 cytokine IL-18. It effectively blocks IL-18 by forming a 1:1 high affinity (Kd=400 pM) complex, exhibiting a very low dissociation rate. We have developed a sandwich ELISA for IL-18BPa and determined its limit of detection (62 pg/ml). Interference by IL-18 and related cytokines, as well as cross reactivity with other IL-18BP isoforms (b, c, and d) were determined. Using this ELISA, we measured serum IL-18BPa in large cohorts of healthy individuals and in septic patients. Serum IL-18BPa in healthy individuals was 2.15+/-0.15 ng/ml (range 0.5-7 ng/ml). In sepsis, the level rose to 21.9+/-1.44 ng/ml (range 4-132 ng/ml). Total IL-18 was measured in the same sera by an electrochemiluminescence assay and free IL-18 was calculated based on the mass action law. Total IL-18 was low in healthy individuals (64+/-17 pg/ml) and most of it ( approximately 85%) was in its free form. Total IL-18 and IL-18BPa were both elevated in sepsis patients upon admission (1.5+/-0.4 ng/ml and 28.6+/-4.5 ng/ml, respectively). At these levels, most of the IL-18 is bound to IL-18BPa, however the remaining free IL-18 is still higher than in healthy individuals. We conclude that IL-18BPa considerably inhibits circulating IL-18 in sepsis. Yet, exogenous administration of IL-18BPa may further reduce circulating IL-18 activity. PMID- 11497495 TI - Effect of ulcerative colitis activity on plasma concentration of transforming growth factor beta1. AB - Enhanced expression of transforming growth factor-beta1 (TGF-beta1) demonstrated in human colonic mucosa of patients with ulcerative colitis (UC), indicates its possible significance in the pathogenesis of this disease. The aim of this study was to evaluate plasma TGF-beta1 concentration in patients with different degrees of colonic mucosal injury, as a possible indicator of ulcerative colitis activity. TGF-beta1 concentration was measured with an enzyme immunoassay (EIA) in plasma of 45 patients with endoscopically confirmed UC. Values observed in UC patients (40.5+/-15.9 ng/ml) were significantly higher than in healthy people (18.3+/-11.6 ng/ml) and higher than in patients with irritable colon syndrome (ICS), (20.5+/-13.6 ng/ml). The highest plasma TGF-beta1 (58.6+/-112.1 ng/ml) was in patients with the severe UC course. TGF-beta1 level analysed in all UC patients revealed significant positive correlation with scored degree of mucosal injury (r=0.396;P<0.01). Among other possible laboratory markers of the disease activity, only C-reactive protein concentration demonstrated significant correlation. Enhanced production of TGF-beta1 can be related to inflammation activity. Measurement of plasma TGF-beta1 may be considered as a biomarker of the disease activity. PMID- 11497496 TI - Isolation, nucleotide sequence and expression of a cDNA encoding feline granulocyte colony-stimulating factor. AB - A cDNA encoding feline granulocyte colony stimulating factor (fG-CSF) was cloned from alveolar macrophages using the reverse transcriptase-polymerase chain reaction. The cDNA is 949 bp in length and encodes a predicted mature protein of 174 amino acids. Recombinant fG-CSF was expressed as a glutathione S-transferase fusion and purified by affinity chromatography. Biological activity of the recombinant protein was demonstrated using the murine myeloblastic cell line GNFS 60, which showed an ED50 for fG-CSF of approximately 2 ng/ml. PMID- 11497497 TI - IL-6 upregulates its own receptor on some human myeloma cell lines. AB - Interleukin 6 (IL-6) is the major survival factor of myeloma cells. In this study, we demonstrate that IL-6, oncostatin M (OSM) and leukemia inhibitory factor (LIF) upregulate membrane IL-6 receptor alpha (IL-6Ralpha) on OPM-2 myeloma cell line at transcriptional level. In OPM-2 cells, IL-6, OSM and LIF induce both signal transducers and activators of transcription (STAT), mitogen activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI 3-K) activation. We show that the cytokine-induced upregulation of IL-6Ralpha can be abolished by a janus kinase (JAK)-2 specific inhibitor, i.e. AG490, suggesting an involvement of the JAK/STAT pathway in this process. Finally, IL-6Ralpha upregulation was also inhibited by wortmannin, an inhibitor of the PI 3-kinase pathway. In conclusion, IL-6 can upregulate its own receptor on OPM-2 cells probably through the JAK/STAT and PI 3-kinase pathways. PMID- 11497498 TI - Cytokines in cervicovaginal washing fluid from patients with cervical neoplasia. AB - Human papillomavirus (HPV) infections play an important role in the development of cervical neoplasia. To get to a better understanding of the role of cytokines in the development of these neoplasias, we analysed the presence of various cytokines in cervicovaginal washings of healthy volunteers (n=22), cervical intraepithelial neoplasia (CIN) patients (n=63) and cervical cancer patients (n=33). IL-12p40, IL-10, TGF-beta1, TNF-alpha and IL-1beta levels were significantly higher in patients with cervical cancer than in controls and CIN patients. The levels of IFN-gamma were not different. Our data demonstrate alterations in the local cervical immune environment in cervical cancer patients. This could have important consequences for the further development of immune modulating therapies and vaccination strategies. PMID- 11497499 TI - A successful 15-year experience in double-dome tip surgery via endonasal approach: nuances and pitfalls. AB - BACKGROUND: Endonasal double-dome techniques provide a reliable method to approach the nasal tip. OBJECTIVES: To review one surgeon's 15-year experience using a graduated method of endonasal double-dome tip surgery including patient selection, intraoperative techniques, and postoperative complications and to emphasize the nuances to achieve symmetry and consistent results. PATIENTS AND METHODS: Three hundred eighty-six patients who had adequate follow-up after undergoing endonasal double-dome tip rhinoplasty. RESULTS: Results at 1 year showed high rates of supratip (94%), dome (96%), and nostril (88%) symmetry. There was a high rate of patient satisfaction with a low rate of revision (7%). CONCLUSION: Endonasal double-dome tip surgery provides the surgeon the ability to achieve consistent results with high patient satisfaction and a low rate of revision. PMID- 11497501 TI - Hair-bearing temporoparietal fascial flap reconstruction of upper lip and scalp defects. AB - BACKGROUND: The temporoparietal fascial flap has proven to be a versatile flap for a broad spectrum of reconstructive problems in the head and neck. The temporoparietal fascial flap is a thin, pliable layer of richly vascularized tissue that may be transferred either pedicled or free and alone or as a carrier of subjacent bone or overlying skin and scalp. OBJECTIVE: To report our experience using a hair-bearing temporoparietal fascial flap for reconstruction in 6 male patients with extensive upper lip and scalp defects, including a discussion of the surgical anatomy and technique. METHODS: Temporoparietal fascial flaps with overlying scalp were used as pedicled and free flaps for the reconstruction of upper lip and scalp defects. RESULTS: All reconstructive results were satisfactory. Oral competence, measured by both speech and mastication performance, was achieved in patients with upper lip defects. Healthy scalp coverage was obtained in patients with local defects. The cosmetic appearance was satisfactory to all patients. CONCLUSIONS: Ideal reconstruction of large upper lip and scalp defects is achieved with local tissue that best mimics the normal face color, texture, and hair-bearing qualities. Hair-bearing temporoparietal fascial flaps possess these characteristics and are an excellent choice for the restoration of function and aesthetics. PMID- 11497500 TI - Effect of botulinum toxin pretreatment on laser resurfacing results: a prospective, randomized, blinded trial. AB - BACKGROUND: Facial laser resurfacing and chemodenervation with botulinum toxin type A are used independently as means of nonsurgical facial rejuvenation. Recent reports in the literature have described combining these 2 therapies, claiming improved and longer-lasting laser resurfacing results. To date, no scientific investigation has been undertaken to prove or disprove this theory. DESIGN: Institutional review board-approved, prospective, randomized, blinded study at university-affiliated outpatient cosmetic surgery offices. INTERVENTION: Patients had one side of their face injected, at specific anatomic subsites (crow's feet, horizontal forehead furrows, and glabellar frown lines), with botulinum toxin 1 week before laser resurfacing. After receiving an injection, patients underwent cutaneous laser exfoliation on both sides of the face with either a carbon dioxide or an erbium dual-mode laser. MAIN OUTCOME MEASURES: Patients' injected (experimental) and noninjected (control) sides were compared after laser resurfacing. Follow-up was documented at 6 weeks, 3 months, and 6 months after laser resurfacing. Subjective evaluation, based on a visual analog scale, was performed in person by a blinded observer. Furthermore, a blinded panel of 3 expert judges (1 facial plastic surgeon, 1 oculoplastic surgeon, and 1 cosmetic dermatologist) graded 35-mm photographs taken during postoperative follow-up visits. RESULTS: Ten female patients were enrolled in the study. A 2-tailed t test showed that all sites that were pretreated with botulinum toxin showed statistically significant improvement (P< or =.05) over the nontreated side, with the crow's feet region showing the greatest improvement. Comparing results between the carbon dioxide and erbium lasers did not result in any statistically significant differences. CONCLUSIONS: Hyperdynamic facial lines, pretreated with botulinum toxin before laser resurfacing, heal in a smoother rhytid-diminished fashion. These results were clinically most significant in the crow's feet region. We recommend pretreatment of movement-associated rhytides with botulinum toxin before laser resurfacing. For optimum results, we further recommend continued maintenance therapy with botulinum toxin postoperatively. PMID- 11497502 TI - Quantitative study of nasal tip support and the effect of reconstructive rhinoplasty. AB - OBJECTIVES: To develop a method to quantify nasal tissue resilience, to establish the normal range for persons without nasal obstruction, and to measure the changes in tissue resilience resulting from standard open rhinoplastic techniques. METHODS: A new device is described that determines nasal tissue resilience. Measurements on the nasal tip were obtained in triplicate at 5 distinct anatomical sites. Normal values (N = 60) were stratified for both sexes into 3 different age groups. Preoperative and postoperative measurements were also obtained in 6 patients who underwent open rhinoplasty for airway obstruction. One patient who underwent intranasal valve repair was included for comparison. All operative patients underwent preoperative and postoperative rhinomanometric measurements. RESULTS: Across all age and sex groups the anterior septal angle is the firmest area of the nasal tip. The mean tissue resilience over the interdomal area and the midcolumella is significantly greater in men than in women. The resilience of the interdomal area exhibits an age effect, with decreasing stiffness over time. The postoperative changes seen correlate well with the placement of structural grafts during rhinoplasty. CONCLUSIONS: Nasal tip support can be quantified. Normative values have been established, which allow one to identify areas of inadequate tip support in persons with nasal obstruction. Alterations in tip support resulting from surgical intervention can be quantified. Open rhinoplasty techniques are an excellent tool to restore deficiencies in nasal tip support. PMID- 11497503 TI - Pediatric mandibular fractures: a free hand technique. AB - BACKGROUND: The treatment of pediatric mandibular fractures is rare, controversial, and complicated by mixed dentition. OBJECTIVES: To determine if open mandibular fracture repair with intraoral and extraoral rigid plate placement, after free hand occlusal and bone reduction, without intermaxillary fixation (IMF), is appropriate and to discuss postoperative advantages, namely, maximal early return of function and minimal oral hygiene issues. PATIENTS: A group of 29 pediatric patients with a mandibular fracture were examined. Twenty pediatric patients (13 males and 7 females) with a mean age of 9 years (age range, 1-17 years) were treated using IMF. All patients were treated by the same surgeon (G.S.). RESULTS: Surgical time for plating was reduced by 1 hour, the average time to place patients in IMF. The patients who underwent open reduction internal fixation without IMF ate a soft mechanical diet by postoperative day 3 compared with postoperative day 16 for those who underwent IMF. Complication rates related to fixation technique were comparable at 20% for those who did not undergo IMF and 33% for those who did. CONCLUSIONS: We believe that free hand reduction is a valuable technique to reduce operative time for pediatric mandibular fractures. It maximizes return to function while minimizing the oral hygiene issues and hardware removal of intermaxillary function. PMID- 11497505 TI - Anthropometric facial analysis of the African American woman. AB - OBJECTIVE: To assess the differences in facial proportions between African American and Caucasian women. Differences within the African American population are sought. DESIGN: Anthropometric survey. PARTICIPANTS: Volunteer sample of African American women (N = 108), aged 18 through 30 years, with African American parents and no previous facial surgery or trauma. INTERVENTION: Photographs and 16 standard anthropometric measurements were taken in concordance with the 9 neoclassical canons. Results were compared with the North American white standard and the neoclassical canons, and an intragroup evaluation was performed. One-way analysis of variance, 99.7% confidence intervals, and t tests were used to test differences for significance. MAIN OUTCOME MEASURES: Anthropometric measures. RESULTS: Compared with white women, the following measurements were found to be significantly different (P<.003) in African American women: special head height was shorter; forehead height II was longer; nose length was shorter; lower face height was longer; height of the calva was shorter; forehead height I was longer; and ear length was shorter. In addition, most horizontal measures were wider, ie, eye-fissure width, nasal width, mouth width, and facial width. The nose and ear have greater angles of inclination. Of the 9 neoclassical canons, the orbital proportion was found to include the most proportional subjects (30.6%), followed by the nasoaural proportion (13.0%) and the nasofacial proportion (9.3%). Subcategorization based on nasal dorsal height yielded the most significantly different measures. CONCLUSIONS: African American female facial anthropometric measures, especially those of the horizontal dimension, differ significantly from those of young white subjects. The average African American woman does not fit the neoclassical standard of facial proportion. PMID- 11497506 TI - Measuring cosmetic facial plastic surgery outcomes: a pilot study. AB - OBJECTIVE: To test 4 previously published outcomes instruments (the Facelift Outcomes Evaluation, the Rhinoplasty Outcomes Evaluation, the Blepharoplasty Outcomes Evaluation, and the Skin Rejuvenation Outcomes Evaluation) in terms of their reliability and validity in assessing patient-related outcomes of surgical intervention. DESIGN: A prospective pilot study of 78 patients in 3 similar private cosmetic surgery centers undergoing a total of 100 face-lift, rhinoplasty, blepharoplasty, and skin rejuvenation procedures. Patients were evaluated at 2 preoperative and 1 postoperative time points and the instruments were analyzed with regard to their test-retest reliability, internal consistency, and responsiveness to change. RESULTS: All 4 outcomes instruments had excellent reliability, consistency, and validity scores. Test-retest reliability was 0.74 to 0.83 (Pearson correlation coefficients), internal consistency scores were.83 to.88 (Cronbach alpha), and responsiveness to change was statistically significant for each instrument tested (P< or =.001). In addition, patients experienced significant quality of life improvement, with overall satisfaction increasing on average from 37% to more than 84% after these procedures. CONCLUSIONS: These 4 instruments are reliable and valid and can be used to accurately assess patient-related satisfaction in studies of face-lift, rhinoplasty, blepharoplasty, and skin resurfacing outcomes. These brief questionnaires provide the cosmetic surgeon with quantitative tools to evaluate otherwise subjective and purely qualitative outcomes and are recommended for use in future prospective studies. PMID- 11497507 TI - Treatment of head and neck melanoma, lentigo maligna subtype: a practical surgical technique. AB - Melanoma with the lentigo maligna histological pattern often provides a significant and difficult challenge to the head and neck surgeon. The lentigo maligna subtype is the most common type of melanoma on the head and neck. This potentially lethal form of cancer is associated with greater nonvisual lesional extension that is often not clinically apparent. Failure to excise the entire lesion results in a higher risk of local recurrence and a poorer prognosis. The staged excision technique described herein results in histological interpretation of 100% of the peripheral margins using formalin-fixed vertical sections. Definitive local excision and soft tissue reconstruction are performed in a subsequent stage, with an assurance that 100% of the peripheral margins have been evaluated and interpreted as free of disease. PMID- 11497508 TI - Laser tissue welding: a biotechnological advance for the future. AB - Laser tissue welding as well as other alternative methods of closure will play a more important role in surgical specialties as laparoscopic, endoscopic, and microsurgical techniques continue to develop. Laser tissue welding uses laser energy to anastomose tissues and is ideally suited for applications in which suturing and stapling is difficult. Recent advances have led to a better understanding of the mechanisms of tissue welding. Additionally, technical achievements including the introduction of protein solders and temperature controlled feedback systems have led to the acceptance of laser tissue welding in clinical medicine. In this article, we describe the history and development of laser tissue welding and review the current and potential applications of this technology. PMID- 11497509 TI - The future of e-facial plastic surgery. PMID- 11497511 TI - Beauty: Vesalius' Fabrica: the marriage of art and anatomy. PMID- 11497512 TI - A piece of my mind: the most primitive sense. PMID- 11497515 TI - Ecstasy experts want realistic messages. PMID- 11497516 TI - NCI to lead palliative care improvements. PMID- 11497517 TI - Racial barriers may hamper diagnosis, care of patients with Alzheimer disease. PMID- 11497522 TI - Systemic vs individualistic approaches to bullying. PMID- 11497524 TI - Persistent pain in nursing home residents. PMID- 11497526 TI - Patient-physician discussions about physician-assisted suicide. PMID- 11497528 TI - Preventing tobacco purchases by underage youths. PMID- 11497529 TI - Sex hormones, amyloid protein, and Alzheimer disease. PMID- 11497531 TI - Association between bone mineral density and serum lipids in men. PMID- 11497532 TI - Recalls and safety alerts involving pacemakers and implantable cardioverter defibrillator generators. AB - CONTEXT: Unanticipated pacemaker and implantable cardioverter-defibrillator (ICD) generator malfunctions sometimes warrant recall by the US Food and Drug Administration (FDA). Despite increasingly frequent device implantation, pacemaker and ICD recalls and safety alerts (advisories) remain poorly characterized. OBJECTIVES: To determine pacemaker and ICD generator advisory rates in the United States, to identify trends in these rates, and to examine their clinical and financial implications. DESIGN AND SETTING: Analysis of weekly FDA Enforcement Reports issued between January 1990 and December 2000 to identify all advisories involving pacemaker or ICD generators in the United States. Recalls and safety alerts involving lead malfunctions were not included. MAIN OUTCOME MEASURES: Number of pacemakers and ICD generators in the United States subject to FDA recall or safety alert in 1990-2000; annual pacemaker and ICD advisory rates in the United States in 1990-2000; and estimated cost of device advisories. RESULTS: During the study period, 52 advisories (median [25th and 75th percentiles], 4 [4 and 7] per year) involving 408 500 pacemakers and 114 645 ICDs (523 145 total devices) were issued. Hardware malfunctions (35 advisories affecting 280 641 devices) and computer errors (10 advisories affecting 216 533 devices) accounted for 95% of device recalls. Implantable cardioverter defibrillators were recalled more frequently than pacemakers (mean [SD], 16.4 [1.6] vs 6.7 [0.8] advisories per 100 person-years; P<.001). Between 1995 and 2000, the annual advisory rate increased for both pacemakers (P for trend <.001) and ICDs (P for trend =.02). An estimated 1.3 million device checks and analyses and 36 187 device replacements resulted from the advisories and cost approximately $870 million. CONCLUSIONS: Pacemaker and ICD recalls and safety alerts occur frequently, affect many patients, and appear to be increasing in number and rate. With the growing number of device implants and expanding indications for device therapy, the number of patients affected by device advisories will likely continue to increase. PMID- 11497533 TI - Status of clinical research in academic health centers: views from the research leadership. AB - CONTEXT: The changing state of the health care system in the United States may be adversely affecting clinical research conducted in academic health centers (AHCs). Few formal data have been gathered about the nature and extent of the problems facing clinical research or the effects of remedies undertaken by AHCs. OBJECTIVES: To assess the perceived quality and health of the clinical research enterprise and to determine challenges and adaptations to current environmental pressures. DESIGN, SETTING, AND PARTICIPANTS: Mailed survey conducted between December 1998 and March 1999 of a subsample of department chairs and senior research administrators (SRAs) in all US medical schools. Of the 712 potential respondents, 478 completed a questionnaire, yielding an overall response rate of 67.1% (64.8% for SRAs and 67.8% for department chairs). MAIN OUTCOME MEASURES: Ratings of overall health/robustness of clinical research, quality of research in 5 domains, extent of challenges to performing research, and sense of urgency in responding to research challenges; formal strategies for research-related tasks and their effects. RESULTS: Slightly more than half (52%) of all respondents rated the health of the clinical research enterprise as good or excellent compared with 63% for nonclinical research (P<.001). Respondents were most likely to rate nonclinical research as high in quality (79%) compared with 70% for phase 3 clinical trials, 67% for translational research, 65% for phase 1 and 2 trials, and 57% for health services research (for all comparisons with nonclinical research, P<.001). Pressure on clinical faculty to see patients was perceived as a moderate-to-large problem for clinical research by the largest percentage of respondents (93%), followed by insufficient clinical revenues (89%), recruiting trained researchers (75%), lack of external support for clinical research (72%), competition from contract research organizations (48%), problems introduced by the institutional review board process (38%), and finding research participants (37%). In total, 81% of respondents considered the challenges facing clinical research in AHCs to be urgent or extremely urgent. CONCLUSIONS: Academic leaders perceive clinical research activities in AHCs to be less healthy, of poorer quality, and facing greater challenges than nonclinical research activities. Many AHCs do not have policies or mechanisms to address challenges facing the clinical research mission. Even among those with such policies, more than half do not believe these policies have had large positive effects. Our findings support the view that the clinical research workforce and infrastructure may need to be expanded and strengthened to keep pace with advances in basic research. PMID- 11497534 TI - Exposure to soy-based formula in infancy and endocrinological and reproductive outcomes in young adulthood. AB - CONTEXT: A large body of evidence documents the role of phytoestrogens in influencing hormone-dependent states. Infants fed soy formula receive high levels of phytoestrogens, in the form of soy isoflavones, during a stage of development at which permanent effects are theoretically possible. However, a paucity of data exists on the long-term effects of infant soy formulas. OBJECTIVE: To examine the association between infant exposure to soy formula and health in young adulthood, with an emphasis on reproductive health. DESIGN, SETTING, AND PARTICIPANTS: Retrospective cohort study conducted from March to August 1999 among adults aged 20 to 34 years who, as infants, participated during 1965-1978 in controlled feeding studies conducted at the University of Iowa, Iowa City (248 were fed soy formula and 563 were fed cow milk formula during infancy). MAIN OUTCOME MEASURES: Self-reported pubertal maturation, menstrual and reproductive history, height and usual weight, and current health, compared based on type of formula exposure during infancy. RESULTS: No statistically significant differences were observed between groups in either women or men for more than 30 outcomes. However, women who had been fed soy formula reported slightly longer duration of menstrual bleeding (adjusted mean difference, 0.37 days; 95% confidence interval [CI], 0.06 0.68), with no difference in severity of menstrual flow. They also reported greater discomfort with menstruation (unadjusted relative risk for extreme discomfort vs no or mild pain, 1.77; 95% CI, 1.04-3.00). CONCLUSIONS: Exposure to soy formula does not appear to lead to different general health or reproductive outcomes than exposure to cow milk formula. Although the few positive findings should be explored in future studies, our findings are reassuring about the safety of infant soy formula. PMID- 11497535 TI - Bone mineral density response to estrogen replacement in frail elderly women: a randomized controlled trial. AB - CONTEXT: Although hormone replacement therapy (HRT) is an established approach for osteoporosis prevention, little is known about the osteoprotective effects of HRT in frail elderly women. OBJECTIVE: To determine whether HRT increases bone mineral density (BMD) in frail elderly women. DESIGN AND SETTING: Randomized, double-blind, placebo-controlled trial conducted in a US university-based research center from September 1995 to August 2000. PARTICIPANTS: Sixty-seven women aged 75 years or older with mild-to-moderate physical frailty. INTERVENTION: Participants were randomly assigned to receive conjugated estrogens, 0.625 mg/d, plus trimonthly medroxyprogesterone acetate, 5 mg/d for 13 days (n = 45), or matching placebo (n = 22), for 9 months. MAIN OUTCOME MEASURES: The primary outcome measure was 9-month change in BMD of the lumbar spine and hip, measured by dual-energy x-ray absorptiometry. Secondary outcomes were changes in markers of bone turnover. RESULTS: Based on intention-to-treat analyses, HRT resulted in significantly larger increases in BMD of the lumbar spine than placebo (mean change, 4.3% vs 0.4%; between-group difference, 3.9%; 95% confidence interval [CI], 3.5%-4.3%) and total hip (mean change, 1.7% vs 0.1%; between-group difference, 1.8%; 95% CI, 1.5%-2.1%). Compared with placebo, HRT resulted in significant decreases in serum bone-specific alkaline phosphatase levels (mean change, -24% vs 6%; between-group difference, -30%; 95% CI, -26% to 33%) and urine N-telopeptide levels (mean change, -48% vs 4%; between-group difference, -52%; 95% CI, -47% to -55%). CONCLUSIONS: In physically frail elderly women, 9 months of HRT significantly increased BMD compared with placebo in clinically important skeletal regions. Further studies are needed to determine whether these osteogenic effects of HRT in elderly women are associated with a reduction in osteoporotic fractures. PMID- 11497536 TI - Comparison of evidence of treatment effects in randomized and nonrandomized studies. AB - CONTEXT: There is substantial debate about whether the results of nonrandomized studies are consistent with the results of randomized controlled trials on the same topic. OBJECTIVES: To compare results of randomized and nonrandomized studies that evaluated medical interventions and to examine characteristics that may explain discrepancies between randomized and nonrandomized studies. DATA SOURCES: MEDLINE (1966-March 2000), the Cochrane Library (Issue 3, 2000), and major journals were searched. STUDY SELECTION: Forty-five diverse topics were identified for which both randomized trials (n = 240) and nonrandomized studies (n = 168) had been performed and had been considered in meta-analyses of binary outcomes. DATA EXTRACTION: Data on events per patient in each study arm and design and characteristics of each study considered in each meta-analysis were extracted and synthesized separately for randomized and nonrandomized studies. DATA SYNTHESIS: Very good correlation was observed between the summary odds ratios of randomized and nonrandomized studies (r = 0.75; P<.001); however, nonrandomized studies tended to show larger treatment effects (28 vs 11; P =.009). Between-study heterogeneity was frequent among randomized trials alone (23%) and very frequent among nonrandomized studies alone (41%). The summary results of the 2 types of designs differed beyond chance in 7 cases (16%). Discrepancies beyond chance were less common when only prospective studies were considered (8%). Occasional differences in sample size and timing of publication were also noted between discrepant randomized and nonrandomized studies. In 28 cases (62%), the natural logarithm of the odds ratio differed by at least 50%, and in 15 cases (33%), the odds ratio varied at least 2-fold between nonrandomized studies and randomized trials. CONCLUSIONS: Despite good correlation between randomized trials and nonrandomized studies-in particular, prospective studies-discrepancies beyond chance do occur and differences in estimated magnitude of treatment effect are very common. PMID- 11497537 TI - Liver enzyme monitoring in patients treated with troglitazone. AB - CONTEXT: Soon after initial marketing in March 1997, troglitazone, the first thiazolidinedione antidiabetic agent, was found to cause life-threatening acute liver failure. The drug was removed from the market in March 2000. OBJECTIVE: To evaluate the effect of US Food and Drug Administration (FDA) risk management efforts, including repeated labeling changes and "Dear Healthcare Professional" letters, on periodic liver enzyme monitoring of patients taking troglitazone. DESIGN, SETTING, AND PARTICIPANTS: Claims data from a large, multistate managed care organization were used to establish 4 cohorts of patients (N = 7603) with at least 90 days of health plan enrollment before first troglitazone prescription during 4 consecutive periods spanning April 1997 to September 1999 and representing 4 progressively stringent liver monitoring recommendations. MAIN OUTCOME MEASURES: Percentage of eligible troglitazone users in each cohort with baseline, monthly (for up to 6 months of continuous use), and complete (baseline and monthly) enzyme monitoring, based on computerized records of laboratory claims. RESULTS: Baseline testing increased from 15% before any FDA monitoring recommendations (cohort 1) to 44.6% following 4 separate FDA interventions (cohort 4; P<.001). In cohort 4, 33.4% of users had follow-up testing after 1 month of therapy, falling to 13% after 5 months of continuous use. In all cohorts, less than 5% received all recommended liver enzyme tests by the third month of continuous use. CONCLUSIONS: The FDA risk management efforts did not achieve meaningful or sustained improvement in liver enzyme testing. Evaluation of the impact of regulatory actions is needed before such actions can be regarded as effective or sufficient. PMID- 11497538 TI - Strategies to decrease tuberculosis in us homeless populations: a computer simulation model. AB - CONTEXT: The rate of tuberculosis (TB) among US homeless persons may be 20 times that of the general adult population. Studies suggest that the majority of urban homeless TB cases are attributable to ongoing transmission of TB. Optimal TB control strategies in both chronically and transiently homeless populations are not known. OBJECTIVE: To examine the effects of TB-control strategies on projected TB cases and deaths in US homeless populations using a computer-based simulation model. DESIGN, SETTING, AND POPULATION: The US general population and a theoretical population of 2 million homeless individuals in 1995 were divided into 18 clinical states based on the risk for or presence of TB and human immunodeficiency virus (HIV) infection in a semi-Markov model. MAIN OUTCOME MEASURES: Prevalence of transiently and chronically homeless individuals with active TB and deaths from TB as a function of public health measures taken to control and eliminate TB, including improvement of treatment effectiveness, improvement in access to treatment, and vaccination with BCG. RESULTS: A 10% increase in access to treatment among homeless persons with active TB produced larger declines in predicted TB cases and deaths after 10 years (cases and deaths among chronically homeless persons decreased 12.5% and 19.8% and among transiently homeless persons dropped 35.9% and 32.4%, respectively) than improvements in the effectiveness of treatment programs (cases and deaths among chronically homeless persons declined 7.2% and 3.1% and among transiently homeless persons dropped 10.9% and 4.1%, respectively). A 10% increase in access to treatment among homeless persons with latent TB infection led to a 6.7% decline in TB among chronically homeless persons and a 5.7% decline among transiently homeless persons, while a 10% improvement in effectiveness of treatment for latent TB infection was associated with declines of 3.0% and 3.3%, respectively. When treatment for latent TB infection was modeled to be the same in vaccinated and nonvaccinated populations, BCG vaccination led to TB case declines of 15.4% and 21.5% in chronically and transiently homeless populations, respectively. CONCLUSIONS: Overcoming barriers faced by homeless individuals in accessing TB treatment programs will be crucial to reducing the burden of TB in this high-risk group. Increased treatment access, improvement in the effectiveness of treatment programs, and BCG vaccination of HIV-negative homeless individuals have the best chance to markedly decrease TB morbidity and mortality. PMID- 11497539 TI - Safety alerts involving device therapy for arrhythmias. PMID- 11497540 TI - Clinical investigators--the endangered species revisited. PMID- 11497545 TI - Availability of HIV care in Central America. AB - Central America is an area with a growing human immunodeficiency virus (HIV) epidemic, but with marked limitations in its health care infrastructure. Estimated adult HIV infection rates range from 0.20% in Nicaragua to 2.01% in Belize. Hospitals and clinicians with experience in HIV care exist mainly, if not only, in capital cities and principal economic centers. Nationally sponsored social security systems in each country consistently offer a wider range of services than do ministry of health systems. Estimated access to the social security system ranges from 0% in Belize and 10% of the population in Honduras to 95% in Costa Rica. Combination antiretroviral therapy is not available through the ministries of health and zidovudine is only sporadically available for prevention of perinatal transmission. Combination therapy is available through the social security system in the countries of Guatemala, Panama, and Costa Rica only. A wide variety of antiretroviral agents are available through private pharmacies in all countries except Belize. With the exception of Costa Ricans, most people with HIV infection in Central America have limited access to HIV specific health services and limited or no access to antiretroviral agents. PMID- 11497546 TI - Description of random Gaussian surfaces by a four-vertex model. AB - A lattice model of random self-affine surfaces is derived using the inverse order of applying the six-vertex model. The well-argued simplification of such an approach results in the four-vertex model. The high numerical efficiency of the four-vertex model is demonstrated by calculating the fractal dimension of contour loops (isolines) of fractional Brownian surfaces as the function of the roughness exponent. PMID- 11497547 TI - Continuously variable survival exponent for random walks with movable partial reflectors. AB - We study a one-dimensional lattice random walk with an absorbing boundary at the origin and a movable partial reflector. On encountering the reflector at site x, the walker is reflected (with probability r) to x-1 and the reflector is simultaneously pushed to x+1. Iteration of the transition matrix, and asymptotic analysis of the probability generating function show that the critical exponent delta governing the survival probability varies continuously between 1/2 and 1 as r varies between 0 and 1. Our study suggests a mechanism for nonuniversal kinetic critical behavior, observed in models with an infinite number of absorbing configurations. PMID- 11497548 TI - Atomic slide puzzle: self-diffusion of an impure atom. AB - In a series of recent papers [Phys. Rev. Lett. 86, 1562 (2001); Nature (London) 408, 665 (2000)] van Gastel and co-workers have presented what may be the first experimental evidence, based on a series of scanning tunnel microscope images, that impure, Indium atoms, embedded into the first, close-packed layer of a Cu(001) surface, are not localized but make concerted, long excursions. Such excursions occur due to continuous reshuffling of the surface following the position exchanges of both impure and host Cu atoms with the naturally occurring surface vacancies. van Gastel and co-workers have also formulated an original lattice-gas type model with asymmetric exchange probabilities, whose numerical solution is in a good agreement with the experimental data. In this paper we propose an exact lattice solution of several versions of this model. PMID- 11497549 TI - Age-dependent dynamics of water in hydrated cement paste. AB - Self-dynamics of water molecules has been studied in hydrated tricalcium silicate as functions of temperature, aging, and in the presence of an additive. A dynamical model taking into account the existence of "immobile water" and "glassy water" has been used to analyze quasielastic neutron spectrometer spectra. We deduced the fraction of the immobile water (p), the stretch exponent (beta), and the average relaxation time (tau) of the glassy water. A quantitative picture for an aspect of the kinetics of the curing process and the structural relaxation parameters of the glassy water have been established. PMID- 11497550 TI - Continuum description of avalanches in granular media. AB - We develop a continuum description of partially fluidized granular flows. Our theory is based on the hydrodynamic equation for the flow coupled with the order parameter equation, which describes the transition between flowing and static components of the granular system. This theory captures important phenomenology recently observed in experiments with granular flows on rough inclined planes [A. Daerr and S. Douady, Nature (London) 399, 241 (1999)]: layer bistability, and transition from triangular avalanches propagating downhill at small inclination angles to balloon-shaped avalanches also propagating uphill for larger angles. PMID- 11497551 TI - Atomic size effects in continuum modeling. AB - Continuum modeling of many physical systems typically assumes that the spatial extent of an atom is small compared to the quantities of interest and can therefore be neglected. We show that this is valid only asymptotically. For many applications of practical interest, the spatial extent of a discrete atom cannot be neglected. We have developed a model for the description of epitaxial growth based on the levelset method, and find that we can accurately predict quantities such as the island densities, if we implement boundary conditions in a region with atomic width, rather than just on a line without any spatial extent. Only in the limit of very large islands and island spacings can this be neglected. PMID- 11497552 TI - Numerical investigation of optically induced director oscillations in nematic liquid crystals. AB - We present a theoretical study of the effects induced by the passage of a linearly polarized light beam through a thin cell of homeotropic nematic liquid crystal. The light is incident at a slightly oblique angle and is polarized perpendicular to the plane of incidence. Experiments in this geometry have revealed a rich variety of complex, time dependent director motion. We solve numerically the director equations for the nematic and compare the results with existing experimental findings. PMID- 11497553 TI - Gyration radius of a circular polymer under a topological constraint with excluded volume. AB - It is nontrivial whether the average size of a ring polymer should become smaller or larger under a topological constraint. Making use of some knot invariants, we numerically evaluate the mean-square radius of gyration for ring polymers having a fixed knot type, where the ring polymers are given by self-avoiding polygons consisting of freely jointed hard cylinders. We obtain plots of the gyration radius versus the number of polygonal nodes for the trivial, trefoil, and figure eight knots. We discuss possible asymptotic behaviors of the gyration radius under the topological constraint. In the asymptotic limit, the size of a ring polymer with a given knot is larger than that of no topological constraint when the polymer is thin, and the effective expansion becomes weak when the polymer is thick enough. PMID- 11497554 TI - Viscoelasticity of dilute solutions of semiflexible polymers. AB - We show using Brownian dynamics simulations and theory how the shear relaxation modulus G(t) of dilute solutions of relatively stiff semiflexible polymers differs qualitatively from that of rigid rods. For chains shorter than their persistence length, G(t) exhibits three time regimes: At very early times, when the longitudinal deformation is affine, G(t) approximately t(-3/4). Over a broad intermediate regime, during which the chain length relaxes, G(t) approximately t( 5/4). At long times, G(t) mimics that of rigid rods. A model of the polymer as an effectively extensible rod with a frequency dependent elastic modulus B(omega) approximately ((i)omega)(3/4) quantitatively describes G(t) throughout the first two regimes. PMID- 11497555 TI - All-optical measurements of the bending rigidity of lipid-vesicle membranes across structural phase transitions. AB - By exploiting the nanometer sensitivity of the confocal response to the position of an in-focus reflecting surface, we measured the bending rigidity of lipid bilayer vesicles with a noninvasive all-optical method. The vesicles were weakly deformed with femtonewton optical force, and the bending rigidity was measured continuously from the L(alpha) through the P(beta(')) to the L(beta(')) phases on the same specimen for the first time. The bending modulus is found to increase by an order of magnitude from the L(alpha) phase to the L(beta(')) phase, as a result of the increasing area-compressibility modulus and bilayer thickness. The dips of bending modulus give precisely the main-transition and pretransition temperatures, which supports the recently proposed chain-melting model of pretransition. PMID- 11497556 TI - Adsorption phenomenon and external field effect on an isotropic liquid containing impurities. AB - The steady-state distribution of ionic charges in a liquid, in the presence of surface adsorption, is determined. The effect of an electric field applied by means of blocking electrodes is considered. The analysis shows that the surface adsorption of ions dissolved in the liquid is responsible for an asymmetry in the electric-field distribution. In the model, the liquid is assumed to be dielectric but to contain impurities. These impurities, by means of a chemical reaction, can bring about ions. The theory takes into account the activation energy for the ionization chemical reaction and the adsorption energy of the ions at the surface. PMID- 11497557 TI - Multiple nonequilibrium steady states for one-dimensional heat flow. AB - A nonequilibrium molecular dynamics model of heat flow in one-dimensional lattices is shown to have multiple steady states for any fixed heat field strength f(e) ranging from zero to a certain positive value. We demonstrate that, depending on the initial conditions, there are at least two possibilities for the system's evolution: (i) formation of a stable traveling wave (soliton), and (ii) chaotic motion throughout the entire simulation. The percentage of the soliton generating trajectories is zero for small field strength f(e), but increases sharply to unity over a critical region of the parameter f(e). PMID- 11497558 TI - Aging and self-organized criticality in driven dissipative systems. AB - We study the noisy dynamics of a close relative to the original sandpile model. Depending on the type of noise and the time scale of observation, we find stationary fluctuations (similar to self-organized criticality) or an aging dynamics with punctuated equilibria, a decreasing rate of events and reset properties qualitatively similar to those of glassy systems, evolution models, and vibrated granular media. PMID- 11497559 TI - Convergence of Monte Carlo simulations to equilibrium. AB - We give two direct, elementary proofs that a Monte Carlo simulation converges to equilibrium provided that appropriate conditions are satisfied. The first proof requires detailed balance while the second is quite general. PMID- 11497560 TI - Effect of colored noise on networks of nonlinear oscillators. AB - We discuss noise-induced pattern formation in different two-dimensional networks of nonlinear oscillators, namely a sequence of biochemical reactions and the Lorenz system. The main focus of the work is on the dependence of these patterns on the correlation time (i.e., the color) of exponentially correlated Gaussian noise. It is seen that in the nonchaotic case, the homogeneity (or average cluster size) goes through a minimum with higher correlation time, while in its chaotic regime the Lorenz system shows a higher degree of synchronization when the correlation time of the noise is increased. In order to elucidate the origin of this phenomenon, the effect of colored noise on the individual oscillator is investigated. It is shown that the specific dependence of the network's homogeneity on the noise correlation time arises from an interplay of the collective behavior and the properties of the single oscillators. PMID- 11497561 TI - Effective rate equations for the overdamped motion in fluctuating potentials. AB - We discuss physical and mathematical aspects of the overdamped motion of a Brownian particle in fluctuating potentials. It is shown that such a system can be described quantitatively by fluctuating rates if the potential fluctuations are slow compared to relaxation within the minima of the potential, and if the position of the minima does not fluctuate. Effective rates can be calculated; they describe the long-time dynamics of the system. Furthermore, we show the existence of a stationary solution of the Fokker-Planck equation that describes the motion within the fluctuating potential under some general conditions. We also show that a stationary solution of the rate equations with fluctuating rates exists. PMID- 11497562 TI - Do strange kinetics imply unusual thermodynamics? AB - We introduce a fractional Fokker-Planck equation (FFPE) for Levy flights in the presence of an external field. The equation is derived within the framework of the subordination of random processes which leads to Levy flights. It is shown that the coexistence of anomalous transport and a potential displays a regular exponential relaxation toward the Boltzmann equilibrium distribution. The properties of the Levy-flight FFPE derived here are compared with earlier findings for a subdiffusive FFPE. The latter is characterized by a nonexponential Mittag-Leffler relaxation to the Boltzmann distribution. In both cases, which describe strange kinetics, the Boltzmann equilibrium is reached, and modifications of the Boltzmann thermodynamics are not required. PMID- 11497563 TI - Fluctuations in the presence of fields: phenomenological Gaussian approximation and a class of thermodynamic inequalities. AB - The fluctuations of thermodynamic systems in the presence of the fields are considered. The approach is of phenomenological nature and developed in a Gaussian approximation. The cases of a magnetizable continuum in a magnetoquasistatic field, as well as the so called discrete systems are used to exemplify the study. In the latter case one finds that the fluctuation estimators depend both on the intrinsic properties of the system and on the characteristics of the environment. Following earlier ideas of one of the authors we present a class of thermodynamic inequalities for the systems investigated in this paper. In the case of two variables these inequalities are nonquantum analogs of the well known quantum Heisenberg "uncertainty" relations. In this context, the fluctuation estimators support the idea that Boltzmann's constant k has the signification of a generic indicator of stochasticity for thermodynamic systems. PMID- 11497564 TI - Rectification by hopping motion through nonsymmetric potentials: local versus global bias. AB - The hopping motion of noninteracting classical particles through nonsymmetric potentials is investigated in the case of alternating local bias. Different to the case of alternating global bias, a resonancelike behavior is observed. The system exhibits rectification effects in the regime of small to medium bias, whereas rectification effects are absent in the strong bias regime. Possible generalizations and applications are outlined. PMID- 11497565 TI - Statistical mechanics of canonical-dissipative systems and applications to swarm dynamics. AB - We develop the theory of canonical-dissipative systems, based on the assumption that both the conservative and the dissipative elements of the dynamics are determined by invariants of motion. In this case, known solutions for conservative systems can be used for an extension of the dynamics, which also includes elements such as the takeup/dissipation of energy. This way, a rather complex dynamics can be mapped to an analytically tractable model, while still covering important features of nonequilibrium systems. In our paper, this approach is used to derive a rather general swarm model that considers (a) the energetic conditions of swarming, i.e., for active motion, and (b) interactions between the particles based on global couplings. We derive analytical expressions for the nonequilibrium velocity distribution and the mean squared displacement of the swarm. Further, we investigate the influence of different global couplings on the overall behavior of the swarm by means of particle-based computer simulations and compare them with the analytical estimations. PMID- 11497566 TI - Energy and pressure of shearing fluids at different state points. AB - Nonequilibrium molecular dynamics simulations are reported at different strain rates (gamma) and thermodynamic state points for a shearing atomic fluid interacting via a Lennard-Jones potential. Our simulations are performed at the Lennard-Jones triple point, a point midway between the triple point and the critical point, and a high point closer to the critical temperature. We find that, for the mid-point and high point, the energy and hydrostatic pressures have strain-rate dependencies of gamma(2), in contrast to the gamma(3/2) dependencies predicted by mode coupling theory. This analytical dependence is consistent with a Taylor series expansion of these quantities as powers of the strain rate tensor. Only at the triple point does the pressure and energy display a nonanalytical dependence on gamma(3/2). PMID- 11497567 TI - Molecular dynamics study of the longitudinal modes in disparate-mass binary liquid mixtures. AB - A series of molecular dynamics simulations of simple liquid binary mixtures of soft spheres with disparate-mass particles were carried out to investigate the origin of the marked differences between the dynamic structure factors of some liquid binary mixtures such as the Li0.7Mg0.3 and Li0.8Pb0.2 alloys. It is shown that the facility for observing peaks associated with fast-propagating modes in the partial Li-Li dynamic structure factor of Li0.8Pb0.2 should be mainly attributed to the structure of this alloy, which is characterized by an incipient ABAB ordering as found in molten salts. The longitudinal dispersion relations at intermediate wave vectors obtained from the longitudinal current spectra are very similar for the two alloys and reflect the existence of both fast-and slow propagating modes of kinetic character associated with light and heavy particles, respectively. The influence of the hardness of the repulsive potential cores as well as the composition of the mixture on the longitudinal collective modes is also discussed. PMID- 11497568 TI - Quantum effects in the dynamics of He probed by inelastic x-ray scattering. AB - Quantum effects in the teraherz dynamics of supercritical 4He have been studied as a function of both density rho and temperature T; they have been characterized through their effects on the second and third spectral moments of the dynamic structure factor S(Q, omega), measured by the inelastic x-ray scattering (IXS) technique. The IXS spectra were collected in the low-Q region below and around the position of the first diffraction peak Q(m), i.e., in a range relatively unusual in this kind of investigation. The measured spectral moments clearly show a departure from their high-T classical expected values. We observe, moreover, that the amplitude of quantum deviations increases slightly with increasing density. This experimental method allows us to extract, even in a region where the dynamics still maintains a collective character, such typical single particle properties as the mean atomic kinetic energy. PMID- 11497569 TI - Diffusion of hard disks and rodlike molecules on surfaces. AB - We study the submonolayer diffusion of hard disks and rodlike molecules on smooth surfaces through numerical simulations and theoretical arguments. We concentrate on the behavior of the various diffusion coefficients as a function of the two dimensional (2D) number density rho in the case where there are no explicit surface-particle interactions. For the hard disk case, we find that while the tracer diffusion coefficient D(T)(rho) decreases monotonically up to the freezing transition, the collective diffusion coefficient D(C)(rho) is wholly determined by the inverse compressibility which increases rapidly on approaching freezing. We also study memory effects associated with tracer diffusion, and present theoretical estimates of D(T)(rho) from the mode-mode coupling approximation. In the case of rigid rods with short-range repulsion and no orientational ordering, we find behavior very similar to the case of disks with the same repulsive interaction. Both D(T)(rho) and the angular diffusion coefficient D(R)(rho) decrease with rho. Also in this case D(C)(rho) is determined by inverse compressibility and increases rapidly close to freezing. This is in contrast to the case of flexible chainlike molecules in the lattice-gas limit, where D(C)(rho) first increases and then decreases as a function of the density due to the interplay between compressibility and mobility. PMID- 11497570 TI - Theory of the generalized dynamic structure factor of polyatomic molecular fluids measured by inelastic x-ray scattering. AB - We describe a theory for the calculation of the generalized dynamic structure factor S(k,omega) as measured by an inelastic x-ray scattering (IXS) experiment on single-component molecular or polyatomic molecular fluids. IXS spectrum of a simple fluid is proportional to the dynamic structure factor of a single species of atom. In the case of a molecular fluid, however, IXS spectrum is a weighted sum of partial dynamic structure factors of pairs of atomic species. The weighting factors are products of the atomic form factors of the pairs. We call this weighted average dynamic structure factor the generalized dynamic structure factor. We extend the formalism of a three effective eigenmode theory (TEE) developed previously for simple fluids to derive an approximate evolution equation for the generalized dynamic structure factor, which can be considered as a generalized hydrodynamic equation for molecular fluids. As examples, we first study the contributions of the partial dynamic structure factor to the generalized dynamic structure factor computed from molecular dynamics simulation of SPC/E model water. We found that the generalized dynamic structure factor of water measured by IXS can be well approximated by the center of mass or the oxygen atom dynamic structure factors. The generalized TEE model was then employed to analyze IXS spectra of nearly fully hydrated dilauroylphosphatidylcholine. The theory is able to fit all of the spectra in the k range from 5 to 32 nm(-1) quantitatively and gives their deconvoluted generalized dynamic structure factors. PMID- 11497571 TI - Adsorption of Lennard-Jones fluid mixture in a planar slit: a perturbative density functional approach. AB - A simple perturbative density functional approach is employed to investigate the adsorption behavior of a model Lennard-Jones fluid confined in a slitlike pore. Adsorption of one-component fluid as well as two-component fluid mixtures in varying pore sizes has been investigated. The results on the density profiles and the excess adsorption obtained from this theory are found to be in overall good agreement with the available computer simulation results. The results are also compared with the same from some recent weighted density based calculations. PMID- 11497572 TI - Numerical investigation of the angle of repose of monosized spheres. AB - This paper presents a numerical study of the angle of repose, a most important macroscopic parameter in characterizing granular materials, by means of a modified distinct element method. Emphasis is given to the effect of variables related to factors such as particle characteristics, material properties, and geometrical constraints. The results show that sliding and rolling frictions are the primary reasons for the formation of a sandpile; particle size and container thickness significantly influence the angle of repose; and the angle of repose is not so sensitive to density, Poisson's ratio, damping coefficient, and Young's modulus. Increasing rolling friction coefficient or sliding friction coefficient increases the angle of repose. Conversely, increasing particle size or container thickness decreases the angle of repose. The underlying mechanisms for these effects are discussed in terms of particle-particle and particle-wall interactions. PMID- 11497573 TI - Effect of vibration on the stability of a gas-fluidized bed of fine powder. AB - We have investigated the effect of vibrations on the stability of gas-fluidized beds of fine powders (particle size approximately 10 microm). The powder is uniformly fluidized by an adjustable gas flow that enables us to control the average solid volume fraction straight phi(0). The fluidized bed is then subjected to a vertical oscillatory motion of controlled amplitude and frequency. The response of the fluidized bed depends essentially on the value of straight phi(0). For straight phi(0)>0.28 the fluidized bed is in a weak solidlike regime, it has a mechanical strength, and particles are static. In this regime vibration causes compaction of the loosely packed bed. For straight phi(0)<0.28 the mechanical strength vanishes and stresses are carried by interstitial gas and collisions. In this fluidlike regime the fluidized bed displays a diffusive dynamics and particles aggregate due to the strong interparticle van der Waals forces. When vibration is applied the powder expands due to the partial disruption of aggregates. However at a critical value of the vibration amplitude A=A(c) either surface (sloshing) or flow (bubbling) instabilities develop. The nucleation of gas bubbles has been correlated to the saturation in particle diffusivity measured elsewhere. The size of the bubbles increases as A is further increased above A(c) and as the vibration frequency is reduced. Moreover, as it should be expected from the predictions of hydrodynamic models, A(c) is independent of cohesivity for particles of the same size and density. PMID- 11497574 TI - Calcium rubidium nitrate: mode-coupling beta scaling without factorization. AB - The fast dynamics of viscous calcium rubidium nitrate is investigated by depolarized light scattering, neutron scattering, and dielectric loss. Fast beta relaxation evolves as in calcium potassium nitrate. The dynamic susceptibilities can be described by the asymptotic scaling law of mode-coupling theory with a shape parameter lambda=0.79; the temperature dependence of the amplitudes extrapolates to T(c) approximately equal 378 K. However, the frequencies of the minima of the three different spectroscopies never coincide, in conflict with the factorization prediction, indicating that the true asymptotic regime is unreachable. PMID- 11497575 TI - Permeability and conductivity for reconstruction models of porous media. AB - The purpose of this paper is to examine representative examples of realistic three-dimensional models for porous media by comparing their geometrical and transport properties with those of the original experimental specimen. The comparison is based on numerically exact evaluations of permeability, formation factor, porosity, specific internal surface, mean curvature, Euler number, local porosity distributions, and local percolation probabilities. The experimental specimen is a three-dimensional computer tomographic image of Fontainebleau sandstone. The three models are examples of physical and stochastic reconstructions for which many of the geometrical characteristics coincide with those of the experimental specimen. We find that in spite of the similarity in the geometrical properties the permeability and formation factor can differ greatly between models and experiment. Our results seem to indicate that the truncation of correlations is responsible for some of these observed discrepancies. A physical reconstruction model by Bakke and Oren [SPEJ 2, 136 (1997)] based on sedimentation, compaction and diagenesis of sandstones yields surprisingly accurate predictions for permeability and conductivity. These findings imply that many of the presently used geometric descriptors of porous media are insufficient for the prediction of transport. PMID- 11497576 TI - Interpretation of small-angle x-ray scattering data from dilute montmorillonite suspensions using a modified Guinier approximation. AB - Smectites are a group of 2:1-layer phyllosilicate minerals that have been extensively studied by small-angle x-ray scattering (SAXS) because of their industrial and environmental significance. In previous studies, a Guinier plot has been used to obtain the radius of gyration of the clay particles, from which geometric information of the particle structure is derived. Using an indirect Fourier transform to treat SAXS data from a dilute montmorillonite suspension, a negative electron contrast at the clay-water interface is observed. This electron inhomogeneity has violated the assumption underlying the application of the Guinier plot, which requires particles to have a uniform electron density. The presence of this inhomogeneity explains the inability of previous studies to correctly determine particle dimensions using the Guinier plot. Using this model of the clay-water interface, a modified Guinier plot has been derived and was experimentally verified. The calculation shows the presence of negative electron contrast at montmorillonite-water interfaces, which is in accordance with the results from the indirect Fourier transform method. This approximation has the potential to predict the geometric information for similar colloids studied by small-angle scattering. PMID- 11497577 TI - Weak and strong dynamic scaling in a one-dimensional driven coupled-field model: effects of kinematic waves. AB - We study the coupled dynamics of the displacement fields in a one-dimensional coupled-field model for drifting crystals, first proposed by Lahiri and Ramaswamy [Phys. Rev. Lett. 79, 1150 (1997)]. We present some exact results for the steady state and the current in the lattice version of the model for a special subspace in the parameter space, within the region where the model displays kinematic waves. We use these results to construct the effective continuum equations corresponding to the lattice model. These equations decouple at the linear level in terms of the eigenmodes. We examine the long-time, large-distance properties of the correlation functions of the eigenmodes by using symmetry arguments, Monte Carlo simulations, and self-consistent mode-coupling methods. For most parameter values, the scaling exponents of the Kardar-Parisi-Zhang equation are obtained. However, for certain symmetry-determined values of the coupling constants the two eigenmodes, although nonlinearly coupled, are characterized by two distinct dynamic exponents. We discuss the possible application of the dynamic renormalization group in this context. PMID- 11497578 TI - Monte Carlo simulations of the nucleation and growth process of colloidal particles. AB - We have examined the effect of the total initial monomer concentration and that of the monomer-monomer attraction energy on the nucleation and growth process of colloidal particles using a reversible aggregation model (Shih-Aksay-Kikuchi model) with the Monte Carlo method. We showed that the equilibrium monomer concentration c(e) exhibited a peak with respect to the total initial monomer concentration c(t). Furthermore, the solution may be divided into three regimes with respect to c(t). In the first regime where the initial monomer concentration was low, all monomers remained as individual monomers in the solution and c(e) increased linearly with c(t). In the second regime where small clusters of monomers began to form, c(e) underwent a peak with respect to c(t). In the third regime where large particles form, c(e) slowly decreased with c(t). Moreover, with increasing monomer-monomer attraction energy, the peak in c(e) moved to a lower c(t) and became sharper. The equilibrium monomer concentration surrounding a particle with respect to particle size was shown to agree with the Kelvin equation, indicating that the model can indeed capture the equilibrium solution physics involving colloidal particles. The peak exhibited in c(e) versus c(t) was manifested as a peak in the monomer concentration versus time under conditions where monomers were gradually fed to the solution. The present simulation is a simulation model for illustrating a peaked solute concentration with respect to time first proposed by LaMer and Dinegar. We further showed that the supersaturation peak in the monomer concentration versus time depended on the feeding rate. The peak height increased with an increasing feeding rate. PMID- 11497579 TI - Stress relaxation of near-critical gels. AB - The time-dependent stress relaxation for a Rouse model of a cross-linked polymer melt is completely determined by the spectrum of eigenvalues of the connectivity matrix. The latter has been computed analytically for a mean-field distribution of cross-links. It shows a Lifshitz tail for small eigenvalues and all concentrations below the percolation threshold, giving rise to a stretched exponential decay of the stress relaxation function in the sol phase. At the critical point the density of states is finite for small eigenvalues, resulting in a logarithmic divergence of the viscosity and an algebraic decay of the stress relaxation function. Numerical diagonalization of the connectivity matrix supports the analytical findings and has furthermore been applied to cluster statistics corresponding to random bond percolation in two and three dimensions. PMID- 11497580 TI - Strong electrostatic interactions in spherical colloidal systems. AB - We investigate spherical macroions in the strong Coulomb coupling regime within the primitive model in salt-free environment. We first show that the ground state of an isolated colloid is naturally overcharged by simple electrostatic arguments illustrated by the Gillespie rule. We furthermore demonstrate that in the strong Coulomb coupling this mechanism leads to ionized states and thus to long range attractions between like-charged spheres. We use molecular dynamics simulations to study in detail the counterion distribution for one and two highly charged colloids for the ground state as well as for finite temperatures. We compare our results in terms of a simple version of a Wigner crystal theory and find excellent qualitative and quantitative agreement. PMID- 11497581 TI - Pattern and wave number selection in magnetic fluids. AB - The formation of patterns of peaks on the free surface of a magnetic fluid subject to a magnetic field normal to the undisturbed interface is investigated theoretically. The relative stability of ridge, square, and hexagon planforms is studied using a perturbative energy minimization procedure. Extending previous studies the finite depth of the fluid layer is taken into account. Moreover, adding the wave number modulus k to the set of variational parameters also the wave number selection problem is addressed. The results are compared with previous investigations and recent experimental findings. PMID- 11497582 TI - Direct observation of crystallization and aggregation in a phase-separating colloid-polymer suspension. AB - The depletion-induced phase separation in a mixture of colloidal particles (PMMA latex) and nonadsorbing polymers [poly(styrene)] in a solvent (mixture of tetralin, cis-decalin, and carbon tetrachloride) was investigated in real space with confocal scanning laser microscopy in the initial, intermediate, and final stage. It was found that the kinetics and the morphology of the phase separation strongly depend on the polymer concentration, and thus on the strength of the depletion-induced attraction between the colloidal particles. At moderate polymer concentrations, crystallization of the PMMA particles is enhanced. At higher polymer concentrations, only aggregation is observed, resulting in amorphous sediments. The aggregation is diffusion-limited or reaction-limited, depending on the polymer concentration. Digital image processing was used to determine the dependence of the aggregation rate and the size of the clusters on the polymer concentration. PMID- 11497583 TI - Periodic surfaces of simple and complex topology: comparison of scattering patterns. AB - We compute scattering patterns for six triply periodic minimal surfaces formed in oil/surfactant/water solutions: Three surfaces of a simple topology, Schwarz P (Im3m), Schwarz D-diamond (Pn3m), and Schoen G-gyroid (Ia3d), and three surfaces of a complex topology, SCN1 (Im3m), CD (Pn3m), and GX6 (Ia3d). We show that in the case of the complex structures, scattering intensity is shifted towards the higher hkl peaks. This might cause their misidentification and wrong estimates about the cell size of the structure. PMID- 11497584 TI - Conductivity of an inverse lyotropic lamellar phase under shear flow. AB - We report conductivity measurements on solutions of closed compact monodisperse multilamellar vesicles (the so-called "onion texture") formed by shearing an inverse lyotropic lamellar L(alpha) phase. The conductivity measured in different directions as a function of the applied shear rate reveals a small anisotropy of the onion structure due to the existence of free oriented membranes. The results are analyzed in terms of a simple model that allows one to deduce the conductivity tensor of the L(alpha) phase itself and the proportion of free oriented membranes. The variation of these two parameters is measured along a dilution line and discussed. The high value of the conductivity perpendicular to the layers with respect to that of solvent suggests the existence of a mechanism of ionic transport through the insulating solvent. PMID- 11497585 TI - Three-dimensional hydrodynamic lattice-gas simulations of domain growth and self assembly in binary immiscible and ternary amphiphilic fluids. AB - We simulate the dynamics of phase assembly in binary immiscible fluids and ternary microemulsions using a three-dimensional hydrodynamic lattice-gas approach. For critical spinodal decomposition we perform the scaling analysis in reduced variables introduced by Jury et al. [Phys. Rev. E 59, R2535 (1999)] and by Bladon et al. [Phys. Rev. Lett. 83, 579 (1999)]. We find a late-stage scaling exponent consistent with the R approximately t(2/3) inertial regime. However, as observed with the previous lattice-gas model of Appert et al. [J. Stat. Phys. 81, 181 (1995)] our data do not fall in the same range of reduced length and time as those of Jury et al. and Bladon et al. For off-critical binary spinodal decomposition we observe a reduction of the effective exponent with decreasing volume fraction of the minority phase. However, the n=1 / 3 Lifshitz-Slyzov Wagner droplet coalescence exponent is not observed. Adding a sufficient number of surfactant particles to a critical quench of binary immiscible fluids produces a ternary bicontinuous microemulsion. We observe a change in scaling behavior from algebraic to logarithmic growth for amphiphilic fluids in which the domain growth is not arrested. For formation of a microemulsion where the domain growth is halted we find that a stretched exponential growth law provides the best fit to the data. PMID- 11497586 TI - Temperature effects on the static and dynamic properties of liquid water inside nanotubes. AB - We report a molecular dynamics simulation study of the behavior of liquid water adsorbed in carbon nanotubes under different thermodynamic conditions. A flexible simple point charged potential has been employed to model internal and intermolecular water interactions. Water-carbon forces are modeled with a Lennard Jones-type potential. We have studied three types of tubes with effective radii ranging from 4.1 to 6.8 A and three temperatures, from 298 to 500 K for a fixed density of 1 g/cm(3). Structure of each thermodynamic state is analyzed through the characterization of the hydrogen-bond network. Time-dependent properties such as the diffusive behavior and molecular vibrational spectra are also considered. We observe the gradual destruction of the hydrogen-bond network together with faster diffusive regimes as temperature increases. A vibrational mode absent in bulk unconstrained water appears in the power spectra obtained from hydrogen velocity autocorrelation functions for all thermodynamic states. That frequency mode should be attributed to confinement effects. PMID- 11497587 TI - Crossover from fragile to strong glassy behavior in kinetically constrained systems. AB - We show the existence of fragile-to-strong transitions in kinetically constrained systems by studying the equilibrium and out-of-equilibrium dynamics of a generic constrained Ising spin chain that interpolates between the symmetric and fully asymmetric cases. We find that for large but finite asymmetry the model displays a crossover from fragile to strong glassy behavior at finite temperature, which is controlled by the asymmetry parameter. The relaxation in the fragile region presents stretched exponential behavior, with a temperature dependent stretching exponent that is predicted. Our results are confirmed by numerical simulations. PMID- 11497588 TI - Molecular dynamics study of the stability of the hard sphere glass. AB - Glassy states have been observed in hard-spherelike colloidal suspensions; however, some recent work suggests that a stable, one-component hard-sphere glass doesn't exist. A possible resolution of this dilemma is that colloidal glass formation results from a small degree of particle polydispersity. In order to investigate this further, we used the molecular-dynamics method to explore the phase behavior of both one- and two-component hard-sphere systems. It was found that the metastable fluid branch of the one-component system ceased to exist at a volume fraction marginally above melting, instead this system always crystallized within a relatively short period of time. Binary systems with a size ratio gamma=0.9 were then used as the simplest approximation to model a polydisperse hard-sphere colloidal system. Here the crystallization process was slowed down dramatically for all volume fractions and the fluid state was maintained for many relaxation times. Indeed, at the lowest volume fraction straight phi=0.55 no sign of crystallization was seen on the simulation time scale. The systems at intermediate volume fractions did eventually crystallize but at the highest volume fraction of straight phi=0.58, a dramatic slowing down in the crystallization process was observed. This is qualitatively in agreement with the experimental results on colloidal suspensions. Using the insight gained from this paper, the reasons behind a polydisperse system forming a stable glass, in contrast to the one-component system, are elucidated. PMID- 11497589 TI - Cascades of popping bubbles along air/foam interfaces. AB - We report image analysis of popping bubbles during the collapsing of two dimensional (2D) and 3D aqueous foams. Although temporal and spatial correlations between successive popping bubbles within avalanches are emphasized, the breaking of a soap film at the air/foam interface seems to be independent of (i) the topology, (ii) the local curvature, and (iii) the size of the popping bubble. Possible mechanisms for cascades of pops are proposed and discussed. PMID- 11497590 TI - Flow of a ferrofluid down a tube in an oscillating magnetic field. AB - The magnetoviscosity of a ferrofluid flowing down a circular tube in the presence of a magnetic field oscillating in the direction of the axis is studied on the basis of ferrohydrodynamics, Maxwell's equations of magnetostatics, and a relaxation equation for the magnetization. Three different relaxation equations, proposed in the literature, are considered. For large amplitude of the oscillating field the three equations lead to different values of the magnetoviscosity. For large magnetic permeability the self-consistent magnetic field generated by the magnetization has significant effect. PMID- 11497591 TI - Bubble propagation in a pipe filled with sand. AB - Granular flow with strong hydrodynamic interactions has been studied experimentally. Experiments have been carried out to study the movement of a single bubble in an inclined tube filled with glass beads and air. A maximum bubble velocity was found at an inclined angle straight theta(m). The density variations in the sand were measured by capacitance measurements, and a decompactification zone was observed just above the bubble when the inclination angle straight theta was larger than straight theta(m). The length of the decompactification front increased with increasing inclination angle and disappeared for angles smaller than straight theta(m). Both pressure and visualization experiments were carried out and compared with the density measurements. PMID- 11497592 TI - Aging dynamics in a colloidal glass. AB - The aging dynamics of colloidal suspensions of Laponite, a synthetic clay, is investigated using dynamic light scattering (DLS) and viscometry after a quench into the glassy phase. DLS allows to follow the diffusion of Laponite particles and reveals that there are two modes of relaxation. The fast mode corresponds to a rapid diffusion of particles within "cages" formed by the neighboring particles. The slow mode corresponds to escape from the cages: its average relaxation time increases exponentially fast with the age of the glass. In addition, the slow mode has a broad distribution of relaxation times, its distribution becoming larger as the system ages. Measuring the concomitant increase of viscosity as the system ages, we can relate the slowing down of the particle dynamics to the viscosity. PMID- 11497593 TI - Molecular dynamics simulation of the fragile glass former orthoterphenyl: a flexible molecule model. II. Collective dynamics. AB - We present a molecular dynamics study of the collective dynamics of a model for the fragile glass former orthoterphenyl. In this model, introduced by Mossa, Di Leonardo, Ruocco, and Sampoli [Phys. Rev. E 62, 612 (2000)], the intramolecular interaction among the three rigid phenyl rings is described by a set of force constants whose value has been fixed in order to obtain a realistic isolated molecule spectrum. The interaction between different molecules is described by a Lennard Jones site-site potential. We study the behavior of the coherent scattering functions F(t)(q,t), considering the density fluctuations of both molecular and phenyl-ring centers of mass; moreover we directly simulate the neutron scattering spectra taking into account both the contributions due to carbon and hydrogens atoms. We compare our results with the main predictions of the mode-coupling theory and with the available coherent neutron scattering experimental data. PMID- 11497594 TI - Weighted density functional theory of the solvophobic effect. AB - We are interested in the spatial density of a molecular fluid in the presence of a solute of arbitrary size and shape. The density functional is written as the sum of a F0[rho(r)] that effectively describes small deviations around the uniform density, plus an energy density part that is responsible for formation of liquid-vapor interface. Using the weighted density approach, we require the density functional to match with several observed properties of the fluid such as equation of state and surface tension. We also show that weighting functions for calculating the weighted density can be obtained from experimental data. Using these elements, we construct a spatial density functional theory of water and apply it to obtain densities and solvation energies of a hard-sphere solute with encouraging results. PMID- 11497595 TI - Kinetics of phase separation in polymer-solvent mixtures. AB - We investigate the kinetics of phase separation in systems with a strong asymmetry in the mobility. This simple model system mimics the segregation kinetics of polymer-solvent mixtures, where the polymer-rich phase forms a low mobility gel. We obtain detailed numerical results for this model, both without and with thermal noise. In the absence of thermal noise, we find that domain growth is slowed down drastically when the low-mobility phase percolates the system. However, thermal noise restores rapid domain growth through a Brownian coalescence mechanism. PMID- 11497596 TI - Macromolecular theory of solvation and structure in mixtures of colloids and polymers. AB - The structural and thermodynamic properties of mixtures of colloidal spheres and nonadsorbing polymer chains are studied within a general two-component macromolecular liquid state approach applicable for all size asymmetry ratios. The dilute limits, when one of the components is at infinite dilution but the other concentrated, are presented and compared to field theory and to models that replace polymer coils with spheres. Whereas the derived analytical results compare well, qualitatively and quantitatively, with mean-field scaling laws where available, important differences from "effective sphere" approaches are found for large polymer sizes or semidilute concentrations. PMID- 11497597 TI - Motion of grains, droplets, and bubbles in fluid-filled nanopores. AB - Molecular dynamics studies of nanometer-sized rigid grains, droplets and bubbles in nanometer-sized pores indicate that the drag force may have a hydrodynamic form if the moving object is dense and small compared to the pore diameter. Otherwise, the behavior is nonhydrodynamic. The terminal speed is insensitive to whether the falling droplet is made of liquid or a solid. The velocity profiles within droplets and bubbles that move in the pore are usually nonparabolic and distinct from those corresponding to individual fluids. The density profiles indicate motional shape distortion of the moving objects. PMID- 11497598 TI - Microstructure evolution and plasticity in Zr-based glass-forming alloys. AB - The microstructure characteristic scale, in partially devitrified Zr-based glass forming alloys, is examined in the light of classical nucleation and growth theory. It is then related to the high-ductility properties of these materials. Stabilization of the icosahedral phase under addition of isoelectronic elements, as evidenced in recent experiments, is discussed on a qualitative basis. PMID- 11497599 TI - Nonlocal diffuse interface theory of thin films and the moving contact line. AB - A nonlocal diffuse interface model is explored using the "lubrication approximation" applicable to thin films. We show the inconsistency of the expansion leading to a nonlinear diffusion model, and solve an untruncated integro-differential mean field equation to compute the equilibrium density profile across the fluid-vapor interface. The disjoining potential and effect of interfacial curvature are computed using approximations compatible with the lubrication approximation. We explore the thick film asymptotics, and find it coinciding with the sharp interface limit. These results are further used for computation of the static contact angle and derivation of an evolution equation for flowing films of dynamic menisci in the lubrication approximation. The structure of the evolution equation is identical to that of the sharp interface theory, but it is free from troublesome divergences near the three-phase contact line. PMID- 11497600 TI - Sharp interface limits of phase-field models. AB - The use of continuum phase-field models to describe the motion of well-defined interfaces is discussed for a class of phenomena that includes order-disorder transitions, spinodal decomposition and Ostwald ripening, dendritic growth, and the solidification of eutectic alloys. The projection operator method is used to extract the "sharp-interface limit" from phase-field models which have interfaces that are diffuse on a length scale xi. In particular, phase-field equations are mapped onto sharp-interface equations in the limits xi(kappa)<<1 and xi(v)/D<<1, where kappa and v are, respectively, the interface curvature and velocity and D is the diffusion constant in the bulk. The calculations provide one general set of sharp-interface equations that incorporate the Gibbs-Thomson condition, the Allen-Cahn equation, and the Kardar-Parisi-Zhang equation. PMID- 11497601 TI - Surface nuclear magnetic relaxation and dynamics of water and oil in macroporous media. AB - Proton nuclear spin-relaxation studies on water- or oil-saturated granular packings and limestone rocks allow estimating surface molecular dynamical parameters. Measurements were performed at various conditions of temperature, magnetic field strengths, and pore size. We show by low field NMR relaxation that changing the amount of surface paramagnetic impurities leads to striking different pore-size dependences of the relaxation times T1 and T2 of liquids in pores. These dependences are well supported by surface-limited or diffusion limited relaxation models. Surface relaxivity parameters rho(1) and rho(2) are deduced from the pore-size dependence in the surface-limited regime. We evidence the frequency and temperature dependence of the surface relaxivity rho(1) by field cycling NMR relaxation and relevant theoretical models. The typical frequency dependence found allows an experimental separation of the surface and bulk microdynamics in porous media. Several surface dynamical parameters, such as diffusion coefficients, activation energies, time of residence, and coefficient of surface affinity, were therefore determined. The methods presented here give a powerful analysis of the surface microdynamics of confined liquids, which can be applied to the study of oil-bearing rocks. PMID- 11497602 TI - Phase ordering in nematic liquid crystals. AB - We study the kinetics of the nematic-isotropic transition in a two-dimensional liquid crystal by using a lattice Boltzmann scheme that couples the tensor order parameter and the flow consistently. Unlike in previous studies, we find that the time dependences of the correlation function, energy density, and number of topological defects obey dynamic scaling laws with growth exponents that, within the numerical uncertainties, agree with the value 1/2 expected from simple dimensional analysis. We find that these values are not altered by the hydrodynamic flow. In addition, by examining shallow quenches, we find that the presence of orientational disorder can inhibit amplitude ordering. PMID- 11497603 TI - Structure of freely suspended chiral smectic films as determined by x-ray reflectivity and optical ellipsometry. AB - We report on a combined x-ray reflectivity and optical ellipsometry study of freely suspended smectic (Sm) films of a chiral liquid crystalline compound with the phase sequence Sm-A-Sm-C(*)(alpha)-Sm-C*-Sm-C(*)(gamma)-Sm-C(*)(A). Using tilt magnitude profiles from x-ray reflectivity as input to model the average optical properties obtained by ellipsometry, tilt direction profiles are also obtained. In this way realistic models can be elaborated for the various types of chiral Sm-C films. We find that the surface layers are more tilted than the interior layers due to surface interactions and finite size effects. For the ferrielectric Sm-C(*)(gamma) phase the tilt direction profile corresponds to a three-layer helix, in agreement with the clock model of chiral Sm-C phases. In thin films the surface interactions suppress the bulk helix structure of the Sm C(*)(alpha) phase and a Sm-C(*)(A)-like structure is formed with an anticlinic layer-by-layer alternation of the tilt directions. PMID- 11497604 TI - Temperature-induced surface transition in nematic liquid crystals oriented by evaporated SiOx. AB - Temperature induced surface transitions observed in nematic liquid crystals oriented by obliquely evaporated SiOx are analyzed. It is shown that the experimental data can be interpreted with a theory for the thermal renormalization of the anisotropic part of the surface energy based on the mean field approximation. By assuming that the nematic scalar order parameter is given by the Maier-Saupe theory, the fit is obtained with only one free parameter. The temperature behaviors of the polar and azimuthal angles are in agreement with the proposed model over the whole temperature range. PMID- 11497605 TI - Bloch wave approach to the optics of crystals. AB - The Bloch wave method is used to find the effective permittivity tensor epsilon of periodic liquid crystals and artificial structures whose period p is short with respect to the light wavelength lambda and whose optical properties are defined by a permittivity field epsilon(r). The main role of the multiple scattering within the periodic medium is evidenced, and very general expressions of epsilon, based on expansions in ascending powers of the ratio p/lambda and of the light wave vector k, are found. Such expansions allow to discuss the general properties of epsilon, to clarify the role of the spatial dispersions, i.e., to separate the part of epsilon explicitly depending on k from its k-independent part, and to find some interesting properties of crystals that are (i) periodic in only one direction, or (ii) locally isotropic. Finally, the limits of validity of the macroscopic model are discussed. Within these limits only a few terms of the power expansions are required, and their expressions are explicitly given. The obtained results are also useful to better understand the macroscopic optical properties of solid crystals. PMID- 11497606 TI - Resonant x-ray scattering study of the antiferroelectric and ferrielectric phases in liquid crystal devices. AB - Resonant x-ray scattering has been used to investigate the interlayer ordering of the antiferroelectric and ferrielectric smectic C* subphases in a device geometry. The liquid crystalline materials studied contain a selenium atom and the experiments were carried out at the selenium K edge allowing x-ray transmission through glass. The resonant scattering peaks associated with the antiferroelectric phase were observed in two devices containing different materials. It was observed that the electric-field-induced antiferroelectric to ferroelectric transition coincides with the chevron to bookshelf transition in one of the devices. Observation of the splitting of the antiferroelectric resonant peaks as a function of applied field also confirmed that no helical unwinding occurs at fields lower than the chevron to bookshelf threshold. Resonant features associated with the four-layer ferrielectric liquid crystal phase were observed in a device geometry. Monitoring the electric field dependence of these ferrielectric resonant peaks showed that the chevron to bookshelf transition occurs at a lower applied field than the ferrielectric to ferroelectric switching transition. PMID- 11497607 TI - Deformation of liquid crystal droplets under the action of an external ac electric field. AB - Deformation of liquid crystal droplets suspended in liquid polymer matrix under the action of external electric field was observed in dependence of ion concentration in such system. Experimental dependence of droplet elongation vs electric field demonstrates nonmonotonous character with increase of ion concentration. The theory that provides the basic agreement with experimental observation is developed. PMID- 11497608 TI - Induction of orientational order in the isotropic phase of a nematic liquid crystal. AB - The orientational properties of an isotropic dense liquid composed by anisotropic molecules, such as a liquid crystal in an isotropic phase, is studied. Using a Langevin-like equation it will be shown that the rotational motion of each molecule can be divided in two elements describing two kinds of physical motion. The first describes the Brownian rotational motion and another the coherent rotation induced by the external fields. It will be shown that, even at the isotropic phase, an order parameter describing the mean degree of alignment of the molecules around a given point can be defined. This order parameter also separates the order coming from the coherent motion from the order generated by the anisotropy in the thermal fluctuations. At the end the proposed model is compared with an experiment and it is shown that the coherent motion is enough to explain the experimental results. PMID- 11497609 TI - Optical determination of flexoelectric coefficients and surface polarization in a hybrid aligned nematic cell. AB - We present an optical study of the influence of both the flexoelectric effect and surface polarization on a hybrid-aligned nematic cell using the half-leaky guided mode technique. Tilt angle profiles, obtained from fits of experimental data (reflectivity curves) taken under applied voltages, are compared with the ones derived by a complete theoretical model. Measurements with an applied alternating voltage allow the evaluation of the anchoring energy by solving the torque balance equation at the planar surface. From measurements with static fields, the sum of flexoelectric coefficients and the surface polarization are determined by numerical solution of Euler-Lagrange equations. PMID- 11497610 TI - Holes and disclinations in hybrid nematic liquid crystal films. AB - Defect patterns in hybrid nematic liquid crystal films of pentylcyanobiphenyl (5CB) on a glycerol surface were studied with polarized transmission and monochromatic reflected light microscopy. We report that disclinations of apparent topological strength zero, stable pairs of +1 and -1 disclinations, and higher strength defects found in these films are stabilized by holes in the liquid crystal film. These holes are produced by a monolayer of surfactant on the glycerol surface. In addition, we describe the topology of the director field in the film during the coalescing of two such holes. PMID- 11497611 TI - Shape model for the molecular interpretation of the flexoelectric effect. AB - A mean-field model for the flexoelectric polarization in nematics is presented, based on a continuous description of director deformations coupled to the molecular degrees of freedom via surface interactions. In such a framework, a consistent picture of the flexoelectric effect is obtained, including both dipolar and quadrupolar contributions, with a realistic account of the molecular characteristics of shape and charge distribution. The method is aimed at establishing a quantitative link between chemical structure and flexoelectric response. It provides numerical estimates of the effect and its temperature dependence and allows the recognition of the relevant molecular features for its emergence. Application to some representative systems, comprising mesogenic molecules and photoisomerizable dopants, is considered; it is shown that simple interpretative schemes can be misleading and a comparison with experimental data is reported. PMID- 11497612 TI - Many-body interactions and correlations in coarse-grained descriptions of polymer solutions. AB - We calculate the two-, three-, four-, and five-body (state-independent) effective potentials between the centers of mass (c.m.'s) of self-avoiding walk polymers by Monte Carlo simulations. For full overlap, these coarse-grained n-body interactions oscillate in sign as (-1)(n), and decrease in absolute magnitude with increasing n. We find semiquantitative agreement with a scaling theory, and use this to discuss how the coarse-grained free energy converges when expanded to arbitrary order in the many-body potentials. We also derive effective density dependent two-body potentials that exactly reproduce the pair-correlations between the c.m. of the self avoiding walk polymers. The density dependence of these pair potentials can be largely understood from the effects of the density independent three-body potential. Triplet correlations between the c.m. of the polymers are surprisingly well, but not exactly, described by our coarse-grained effective pair potential picture. In fact, we demonstrate that a pair potential cannot simultaneously reproduce the two- and three-body correlations in a system with many-body interactions. However, the deviations that do occur in our system are very small, and can be explained by the direct influence of three-body potentials. PMID- 11497613 TI - Effects of a nanoscopic filler on the structure and dynamics of a simulated polymer melt and the relationship to ultrathin films. AB - We perform molecular dynamics simulations of an idealized polymer melt surrounding a nanoscopic filler particle. We show that the glass transition temperature T(g) of the melt can be shifted to either higher or lower temperatures by tuning the interactions between polymer and filler. A gradual change of the polymer dynamics approaching the filler surface causes the change in the glass transition. We also find that polymers close to the surface tend to be elongated and flattened. Our findings show a strong similarity to those obtained for ultrathin polymer films. PMID- 11497614 TI - Semiempirical prediction of protein folds. AB - We introduce a semiempirical approach to predict ab initio expeditious pathways and native backbone geometries of proteins that fold under in vitro renaturation conditions. The algorithm is engineered to incorporate a discrete codification of local steric hindrances that constrain the movements of the peptide backbone throughout the folding process. Thus, the torsional state of the chain is assumed to be conditioned by the fact that hopping from one basin of attraction to another in the Ramachandran map (local potential energy surface) of each residue is energetically more costly than the search for a specific (Phi, Psi) torsional state within a single basin. A combinatorial procedure is introduced to evaluate coarsely defined torsional states of the chain defined "modulo basins" and translate them into meaningful patterns of long range interactions. Thus, an algorithm for structure prediction is designed based on the fact that local contributions to the potential energy may be subsumed into time-evolving conformational constraints defining sets of restricted backbone geometries whereupon the patterns of nonbonded interactions are constructed. The predictive power of the algorithm is assessed by (a) computing ab initio folding pathways for mammalian ubiquitin that ultimately yield a stable structural pattern reproducing all of its native features, (b) determining the nucleating event that triggers the hydrophobic collapse of the chain, and (c) comparing coarse predictions of the stable folds of moderately large proteins (N approximately 100) with structural information extracted from the protein data bank. PMID- 11497615 TI - Nonlinear focusing of DNA macromolecules. AB - The present paper reports the nonlinear electrophoretic focusing techniques developed after an original idea by Chacron and Slater [Phys. Rev. E 56, 3436 (1997)]. Focusing of DNA molecules is achieved in an alternating nonuniform electric field, created in a wedge gel with hyperbolic boundaries. The fractions separated on such a wedge retained their rectilinear shape during the electrophoresis. Experiments with gel electrophoresis confirm the possibility of a noticeable nonlinear focusing of DNA molecules. PMID- 11497616 TI - Effects of anatomical constraints on tumor growth. AB - Competition for available nutrients and the presence of anatomical barriers are major determinants of tumor growth in vivo. We extend a model recently proposed to simulate the growth of neoplasms in real tissues to include geometrical constraints mimicking pressure effects on the tumor surface induced by the presence of rigid or semirigid structures. Different tissues have different diffusivities for nutrients and cells. Despite the simplicity of the approach, based on a few inherently local mechanisms, the numerical results agree qualitatively with clinical data (computed tomography scans of neoplasms) for the larynx and the oral cavity. PMID- 11497617 TI - Curved tails in polymerization-based bacterial motility. AB - The curved actin "comet-tail" of the bacterium Listeria monocytogenes is a visually striking signature of actin polymerization-based motility. Similar actin tails are associated with Shigella flexneri, spotted-fever Rickettsiae, the Vaccinia virus, and vesicles and microspheres in related in vitro systems. We show that the torque required to produce the curvature in the tail can arise from randomly placed actin filaments pushing the bacterium or particle. We find that the curvature magnitude determines the number of actively pushing filaments, independent of viscosity and of the molecular details of force generation. The variation of the curvature with time can be used to infer the dynamics of actin filaments at the bacterial surface. PMID- 11497618 TI - Neural-network analysis of the vibrational spectra of N-acetyl L-alanyl N'-methyl amide conformational states. AB - Density-functional theory (DFT) calculations utilizing the Becke 3LYP hybrid functional have been carried out for N-acetyl L-alanine N'-methylamide and examined with respect to the effect of water on the structure, the vibrational frequencies, vibrational absorption (VA), vibrational circular dichroism (VCD), Raman spectra, and Raman optical activity (ROA) intensities. The large changes due to hydration in the structures, and the relative stability of the conformer, reflected in the VA, VCD, Raman spectra, and ROA spectra observed experimentally, are reproduced by the DFT calculations. A neural network has been constructed for reproducing the inverse scattering data (we infer the structural coordinates from spectroscopic data) that the DFT method could produce. The purpose of the network has also been to generate the large set of conformational states associated with each set of spectroscopic data for a given conformer of the molecule by interpolation. Finally the neural network performances are used to monitor a sensitivity analysis of the importance of secondary structures and the influence of the solvent. The neural network is shown to be good in distinguishing the different conformers of the small alanine peptide, especially in the gas phase. PMID- 11497619 TI - Simulated ecology-driven sympatric speciation. AB - We introduce a multilocus genetically acquired phenotype, submitted to mutations and with selective value, in an age-structured model for biological aging. This phenotype describes a single-trait effect of the environment on an individual, and we study the resulting distribution of this trait among the population. In particular, our simulations show that the appearance of a double phenotypic attractor in the ecology induces the emergence of a stable polymorphism, as observed in the Galapagos finches. In the presence of this polymorphism, the simulations generate short-term speciation, when mating preferences are also allowed to suffer mutations and acquire selective value. PMID- 11497620 TI - Osmotic properties of DNA: critical evaluation of counterion condensation theory. AB - The osmotic coefficient of B-DNA in water may, in dilute solutions, deviate by as much as 100% from predictions based on a simple line-charge "counterion condensation" theory. In contrast, a cell model description of the ionic atmosphere near a cylindrical polyelectrolyte predicts osmotic properties that are in surprisingly good harmony with all available experimental findings over a wide range of DNA concentrations. We argue that the neglect of molecular features, such as finite radius, makes line-charge condensation theory inapplicable at all but impractically low polyelectrolyte concentrations. PMID- 11497621 TI - Active membrane fluctuations studied by micropipet aspiration. AB - We present a detailed analysis of the micropipet experiments recently reported by J-B. Manneville et al., [Phys. Rev. Lett. 82, 4356 (1999)], including a derivation of the expected behavior of the membrane tension as a function of the areal strain in the case of an active membrane, i.e., containing a nonequilibrium noise source. We give a general expression, which takes into account the effect of active centers both directly on the membrane and on the embedding fluid dynamics, keeping track of the coupling between the density of active centers and the membrane curvature. The data of the micropipet experiments are well reproduced by our expressions. In particular, we show that a natural choice of the parameters quantifying the strength of the active noise explains both the large amplitude of the observed effects and its remarkable insensitivity to the active-center density in the investigated range. PMID- 11497622 TI - Information transfer with rate-modulated Poisson processes: a simple model for nonstationary stochastic resonance. AB - Stochastic resonance in a simple model of information transfer is studied for sensory neurons and ensembles of ion channels. An exact expression for the information gain is obtained for the Poisson process with the signal-modulated spiking rate. This result allows one to generalize the conventional stochastic resonance (SR) problem (with periodic input signal) to the arbitrary signals of finite duration (nonstationary SR). Moreover, in the case of a periodic signal, the rate of information gain is compared with the conventional signal-to-noise ratio. The paper establishes the general nonequivalence between both measures notwithstanding their apparent similarity in the limit of weak signals. PMID- 11497623 TI - Effects of thermal fluctuation and the receptor-receptor interaction in bacterial chemotactic signaling and adaptation. AB - Bacterial chemotaxis is controlled by the conformational changes of the receptors in response to the change of the ambient chemical concentration. In a statistical mechanical approach, the signaling due to the conformational changes is a thermodynamic average quantity, dependent on the temperature and the total energy of the system, including both ligand-receptor interaction and receptor-receptor interaction. This physical theory suggests to biology an understanding of cooperation in ligand binding and receptor signaling problems. How much experimental support of this approach can be obtained from the currently available data? What are the parameter values? What is the practical information for experiments? Here we make comparisons between the theory and recent experimental results. Although currently comparisons can only be semiquantitative or qualitative, consistency is clearly shown. The theory also helps to sort a variety of data. PMID- 11497624 TI - Wetting of biological lipids on aqueous substrates. AB - We study the dynamics and final wetting state of skin lipids on water and brine by fluorescence microscopy and ellipsometry. When a lipid droplet is brought into contact with the water surface, a lipid wetting film spreads out rapidly by a Marangoni effect. Subsequently, this film undergoes a dewetting instability. However, the final equilibrium is not partial wetting. The film breaks up into droplets with a mesoscopic (approximately 50 A) film in between. These observations result from a subtle interplay between short- and long-range forces: surfactants naturally present in the lipids favor wetting, while the van der Waals forces oppose it. In addition, this reveals the likely organization of the hydrolipid film that covers and protects the skin. PMID- 11497625 TI - Theoretical model of neuronal population coding of stimuli with both continuous and discrete dimensions. AB - In a recent study, the initial rise of the mutual information between the firing rates of N neurons and a set of p discrete stimuli has been analytically evaluated, under the assumption that neurons fire independently of one another to each stimulus and that each conditional distribution of firing rates is Gaussian. Yet real stimuli or behavioral correlates are high dimensional, with both discrete and continuously varying features. Moreover, the Gaussian approximation implies negative firing rates, which is biologically implausible. Here, we generalize the analysis to the case where the stimulus or behavioral correlate has both a discrete and a continuous dimension, like orientation and shape could be in a visual stimulus, or type and direction in a motor action. The functional relationship between the firing patterns and the continuous correlate is expressed through the tuning curve of the neuron, using two different parameters to modulate its width and its flatness. In the case of large noise, we evaluate the mutual information up to the quadratic approximation as a function of population size. We also show that in the limit of large N and assuming that neurons can discriminate between continuous values with a resolution Delta(theta), the mutual information grows to infinity like ln(1/Delta(theta)) when Delta(theta) goes to zero. Then we consider a more realistic distribution of firing rates, truncated at zero, and we prove that the resulting correction, with respect to the Gaussian firing rates, can be expressed simply as a renormalization of the noise parameter. Finally, we demonstrate the effect of averaging the distribution across the discrete dimension, evaluating the mutual information only with respect to the continuously varying correlate. PMID- 11497626 TI - Stochastic resonance in a Hodgkin-Huxley neuron in the absence of external noise. AB - We study numerically nonlinear responses of a periodically forced Hodgkin-Huxley neuron. The coherence of the system in the absence of external noise, namely, the "intrinsic stochastic resonance," is evidenced by the multimodal aperiodic firing pattern, a bell-shaped curve in the signal-to-noise ratio, and the statistical features of the mean firing rate. The subthreshold intrinsic oscillations enhance the signal transduction in a manner different from that in models studied previously. PMID- 11497627 TI - Small world effects in evolution. AB - For asexual organisms point mutations correspond to local displacements in the genotypic space, while other genotypic rearrangements represent long-range jumps. We investigate the spreading properties of an initially homogeneous population in a flat fitness landscape, and the equilibrium properties on a smooth fitness landscape. We show that a small-world effect is present: even a small fraction of quenched long-range jumps makes the results indistinguishable from those obtained by assuming all mutations equiprobable. Moreover, we find that the equilibrium distribution is a Boltzmann one, in which the fitness plays the role of an energy, and mutations that of a temperature. PMID- 11497628 TI - Patchy environment as a factor of complex plankton dynamics. AB - We study the role of the diffusive interaction in plankton dynamics in a patchy environment. We use a minimal reaction-diffusion model of the nutrient-plankton fish food chain to simulate the diffusive interaction between fish-populated and fish-free habitats. We show that such interaction can give rise to spatiotemporal plankton patterns. The plankton dynamics depend on the fish predation rate and can exhibit both regular and chaotic behavior. We show that limit cycle and chaotic attractor coexist in the system. The entire basin of attraction of the limit cycles is found to be riddled with "holes" leading to the competitive chaotic attractors. The chaotic dynamics is typical of a wide range of the fish predation rates. PMID- 11497629 TI - Non-Debye dielectric relaxation in biological structures arises from their fractal nature. AB - What differentiates biological tissues from one another, thereby allowing their accomplishment of a physiological function, is their organization at supracellular and cellular levels. We developed general dielectric models for Cantorian (or treelike) fractal networks of transmission lines that mimic supracellular organization in numerous biological tissues and tissue surfaces, and which are compatible with both in vitro and in vivo measuring techniques. By varying a set of adjustable physical and geometrical parameters pertaining to the structure, we could numerically reproduce a variety of dielectric dispersion curves-most of them of a composite type-that suitably described experimental data from relatively organized biological tissues. We therefore conclude that the well documented non-Debye dielectric behavior of biological structures reflects their self-similar architecture. PMID- 11497631 TI - Nonequilibrium steady-state circulation and heat dissipation functional. AB - A nonequilibrium steady-state (NESS), different from an equilibrium, is sustained by circular balance rather than detailed balance. The circular fluxes are driven by energy input and heat dissipation, accompanied by a positive entropy production. Based on a Master equation formalism for NESS, we show the circulation is intimately related to the recently studied Gallavotti-Cohen symmetry of heat dissipation functional, which in turn suggests a Boltzmann's formulalike relation between rate constants and energy in NESS. Expanding this unifying view on NESS to diffusion is discussed. PMID- 11497630 TI - Stationary solutions of linear stochastic delay differential equations: applications to biological systems. AB - Recently, Kuchler and Mensch [Stochastics Stochastics Rep. 40, 23 (1992)] derived exact stationary probability densities for linear stochastic delay differential equations. This paper presents an alternative derivation of these solutions by means of the Fokker-Planck approach introduced by Guillouzic [Phys. Rev. E 59, 3970 (1999); 61, 4906 (2000)]. Applications of this approach, which is argued to have greater generality, are discussed in the context of stochastic models for population growth and tracking movements. PMID- 11497632 TI - Zero-frequency critical bulk viscosity: is the amplitude ratio truly universal? AB - It was shown by Onuki that the zero-frequency bulk viscosity is associated with a universal amplitude ratio that was calculated to be around 0.10. We show that the sound attenuation data can be used to extract a value for this universal number and we find this number to be around 0.18, reasonably close to Onuki's estimate. However, we argue that a reconsideration of this amplitude ratio shows that this ratio is not truly universal. It has a logarithmic correction instead. PMID- 11497633 TI - Rotational tracer diffusion in binary colloidal sphere mixtures. AB - We demonstrate that tracer/host size asymmetry and electrostatic interactions strongly affect rotational self-diffusion in binary mixtures of charged colloidal tracer and host spheres. Tracer diffusion coefficients, measured with time resolved phosphorescence anisotropy, are compared with calculations of rotational diffusion including two- and three-particle hydrodynamic interactions. We also show that the inverse dependence of the rotational diffusion coefficient on the suspension viscosity is approached only at large size ratios. PMID- 11497634 TI - Properties of surface wave trains excited by mass transfer through a liquid surface. AB - In annular containers, various traveling periodic surface wave trains are generated in liquid layers during the absorption process of a miscible surface active substance out of the vapor phase. Single and counter-rotating wave trains are observed. We here report on waves found to be dispersion-free associated to mostly longitudinal, dilational surface-tension-gradient-driven motions. We report on interactions of the wave crests and on modulations that lead to wave number changes of the wave trains. The wave interactions show behavior typically known for solitons. PMID- 11497635 TI - Casimir torques between anisotropic boundaries in nematic liquid crystals. AB - Fluctuation-induced interactions between anisotropic objects immersed in a nematic liquid crystal are shown to depend on the relative orientation of these objects. The resulting long-range "Casimir" torques are explicitly calculated for a simple geometry where elastic effects are absent. Our study generalizes previous discussions restricted to the case of isotropic walls, and leads to new proposals for experimental tests of Casimir forces and torques in nematics. PMID- 11497636 TI - Prediction of stable walking for a toy that cannot stand. AB - Previous experiments [M. J. Coleman and A. Ruina, Phys. Rev. Lett. 80, 3658 (1998)] showed that a gravity-powered toy with no control and that has no statically stable near-standing configurations can walk stably. We show here that a simple rigid-body statically unstable mathematical model based loosely on the physical toy can predict stable limit-cycle walking motions. These calculations add to the repertoire of rigid-body mechanism behaviors as well as further implicating passive dynamics as a possible contributor to stability of animal motions. PMID- 11497637 TI - Nuclear-spin relaxation induced by shape fluctuations in membrane vesicles. AB - Nuclear-spin relaxation rates resulting from shape fluctuations of unilamellar quasispherical vesicles are calculated. We show that in the kHz range these fluctuations yield-in contrast to previous conclusions on planar membranes - a relaxation rate proportional to the inverse Larmor frequency and provide direct information on the bending rigidity of membranes. PMID- 11497638 TI - Giant strongly connected component of directed networks. AB - We describe how to calculate the sizes of all giant connected components of a directed graph, including the strongly connected one. In particular, the World Wide Web is a directed network. The results are obtained for graphs with statistically uncorrelated vertices and an arbitrary joint in and out-degree distribution P(k(i),k(o)). We show that if P(k(i),k(o)) does not factorize, the relative size of the giant strongly connected component deviates from the product of the relative sizes of the giant in- and out-components. The calculations of the relative sizes of all the giant components are demonstrated using the simplest examples. We explain that the giant strongly connected component may be less resilient to random damage than the giant weakly connected one. PMID- 11497639 TI - Clustering and preferential attachment in growing networks. AB - We study empirically the time evolution of scientific collaboration networks in physics and biology. In these networks, two scientists are considered connected if they have coauthored one or more papers together. We show that the probability of a pair of scientists collaborating increases with the number of other collaborators they have in common, and that the probability of a particular scientist acquiring new collaborators increases with the number of his or her past collaborators. These results provide experimental evidence in favor of previously conjectured mechanisms for clustering and power-law degree distributions in networks. PMID- 11497640 TI - Tunneling mechanism due to chaos in a complex phase space. AB - We have revealed that the barrier-tunneling process in nonintegrable systems is strongly linked to chaos in complex phase space by investigating a simple scattering map model. The semiclassical wave function reproduces complicated features of tunneling perfectly and it enables us to solve all the reasons why those features appear in spite of absence of chaos on the real plane. Multigeneration structure of manifolds, which is the manifestation of complex domain homoclinic entanglement created by complexified classical dynamics, allows a symbolic coding and it is used as a guiding principle to extract dominant complex trajectories from all the semiclassical candidates. PMID- 11497641 TI - Chaotic self-trapping of a weakly irreversible double Bose condensate. AB - We analyze the dynamics of a weakly open Bose-Einstein condensate trapped in a double-well potential. Close to the self-trapping bifurcation, numerical simulations of the weakly irreversible one-dimensional Gross Pitaevskii equation reveal chaotic behaviors. A two-mode model is used to derive amplitude equations describing the complex dynamic of the condensate. PMID- 11497642 TI - Robust chaos in smooth unimodal maps. AB - Robust chaos is defined by the absence of periodic windows and coexisting attractors in some neighborhood of the parameter space. It has been conjectured that robust chaos cannot occur in smooth systems [E. Barreto, B. Hunt, and C. Grebogi, Phys. Rev. Lett. 78, 4561 (1997); 80, 3049 (1998)]. Contrary to this conjecture, we describe a general procedure for generating robust chaos in smooth unimodal maps. PMID- 11497643 TI - Effects of nonionizing prepulses in high-intensity laser-solid interactions. AB - We present theoretical and experimental evidence that nonionizing prepulses with intensities as low as 10(8)-10(9) W/cm(2) can substantially alter high intensity laser-solid interactions. We show that prepulse-heating and vaporization of the target can lead to a preformed plasma once the vapor is ionized by the rising edge of the high-intensity pulse. Our results indicate that peak prepulse intensity is not the only important parameter to consider in determining preformed plasma thresholds, and that a more comprehensive analysis of the prepulse duration and the target material is required. PMID- 11497644 TI - Self-excited vertical oscillations in an rf-discharge dusty plasma. AB - A self-excited vertical oscillation of dust grains in the sheath region of an rf discharge plasma has been observed. The variation of amplitude with pressure and input power was measured. Dramatic increase of oscillation amplitude was found for pressures below 4.5 Pa. Possible excitation mechanisms are considered. PMID- 11497645 TI - Long-time fluctuations in a dynamical model of stock market indices. AB - Financial time series typically exhibit strong fluctuations that cannot be described by a Gaussian distribution. Recent empirical studies of stock market indices examined whether the distribution P(r) of returns r(tau) after some time tau can be described by a (truncated) Levy-stable distribution L(alpha)(r) with some index 02, namely, beyond the range of Levy-stable distributions. Our results are in agreement with both empirical studies and reconcile the apparent disagreement between their results. PMID- 11497646 TI - Dynamics of a spin-1 Ising system in the neighborhood of equilibrium states. AB - The dynamics of a spin-1 Ising system containing biquadratic interactions near equilibrium states is formulated by the method of thermodynamics of irreversible processes. From the expression for the entropy production, generalized forces and fluxes are determined. The kinetic equations are obtained by introducing kinetic coefficients that satisfy the Onsager relation. By solving these equations a set of relaxation times is calculated and examined for temperatures near the phase transition temperatures, with the result that one of the relaxation times approaches infinity near the second-order phase transition temperature on either side, whereas it is sharply cusped at the first-order phase transition temperature. On the other hand, the other relaxation time has a cusp at the second-order phase transition temperature but displays a different behavior at the first-order phase transition, just a jump discontinuity. The behavior of both relaxation times is also investigated at the tricritical point. Moreover, the phase transition behaviors of the relaxation times are also obtained analytically via the critical exponents. Results are compared with conventional kinetic theory in the random-phase or generalized molecular-field approximation and a very good overall agreement is found. PMID- 11497647 TI - Multifractal random walk. AB - We introduce a class of multifractal processes, referred to as multifractal random walks (MRWs). To our knowledge, it is the first multifractal process with continuous dilation invariance properties and stationary increments. MRWs are very attractive alternative processes to classical cascadelike multifractal models since they do not involve any particular scale ratio. The MRWs are indexed by four parameters that are shown to control in a very direct way the multifractal spectrum and the correlation structure of the increments. We briefly explain how, in the same way, one can build stationary multifractal processes or positive random measures. PMID- 11497648 TI - Variations of the asset prices. AB - The empirical established non-Gaussian behavior of asset price fluctuations is studied using an analytical approach. The analysis is based on a nonlinear Fokker Planck equation with a self-organized feedback-coupling term, devised as a fundamental model for price dynamics. The evidence, and the analytical form of the memory term, are discussed in the context of statistical physics. It will be suggested that the memory term in leading order offers a power law dependence with an exponent straight theta. The stationary solution of the probability density leads asymptotically to a truncated Levy distribution, the characteristic exponent beta of which is related to the exponent straight theta by beta=3/theta 1. The empirical data can be reproduced by theta approximately 5/4. PMID- 11497649 TI - Ordering dynamics of the driven lattice-gas model. AB - The evolution of a two-dimensional driven lattice-gas model is studied on an LxxL(y) lattice. Scaling arguments and extensive numerical simulations are used to show that starting from random initial configuration the model evolves via two stages: (a) an early stage in which alternating stripes of particles and vacancies are formed along the direction y of the driving field, and (b) a stripe coarsening stage, in which the number of stripes is reduced and their average width increases. The number of stripes formed at the end of the first stage is shown to be a function of L(x)/L(straight phi)(y), with straight phi approximately 0.2. Thus, depending on this parameter, the resulting state could be either single or multistriped. In the second, stripe coarsening stage, the coarsening time is found to be proportional to L(y), becoming infinitely long in the thermodynamic limit. This implies that the multistriped state is thermodynamically stable. The results put previous studies of the model in a more general framework. PMID- 11497650 TI - Quantum vortices in systems obeying a generalized exclusion principle. AB - The paper deals with a planar particle system obeying a generalized exclusion principle (EP) and governed, in the mean field approximation, by a nonlinear Schrodinger equation. We show that the EP involves a mathematically simple and physically transparent mechanism, which allows the genesis of quantum vortices in the system. We obtain in a closed form the shape of the vortices and investigate its main physical properties. PMID- 11497651 TI - Dynamics and critical behavior of the q model. AB - The q model, a random walk model rich in behavior and applications, is investigated. We introduce and motivate the q model via its application proposed by Coppersmith et al. to the flow of stress through granular matter at rest. For a special value of its parameters the q model has a critical point that we analyze. To characterize the critical point we imagine that a uniform load has been applied to the top of the granular medium and we study the evolution with depth of fluctuations in the distribution of load. Close to the critical point explicit calculation reveals that the evolution of load exhibits scaling behavior analogous to thermodynamic critical phenomena. The critical behavior is remarkably tractable: the harvest of analytic results includes scaling functions that describe the evolution of the variance of the load distribution close to the critical point and of the entire load distribution right at the critical point, values of the associated critical exponents, and determination of the upper critical dimension. These results are of intrinsic interest as a tractable example of a random critical point. Of the many applications of the q model, the critical behavior is particularly relevant to network models of river basins, as we briefly discuss. Finally we discuss circumstances under which quantum network models that describe the surface electronic states of a quantum Hall multilayer can be mapped onto the classical q model. For mesoscopic multilayers of finite circumference the mapping fails; instead a mapping to a ferromagnetic supersymmetric spin chain has proved fruitful. We discuss aspects of the superspin mapping and give an elementary derivation of it making use of operator rather than functional methods. PMID- 11497652 TI - Diffusion-limited reaction in the presence of n traps. AB - We solve the problem of a one-dimensional array of n imperfect traps. These traps are physically represented by small regions of space (in the one-dimensional version we discuss here these traps are represented by small sections of the x axis) with a smaller diffusion constant than that outside them. Small physical particles of one kind diffuse outside and through these small sections. In this work we investigate the changes of the particles density incurred by the presence of these traps. We also check how this density behaves when the density of traps becomes very large. PMID- 11497653 TI - Efficient transition path sampling for nonequilibrium stochastic dynamics. AB - The transition path sampling methodology is adapted to the efficient sampling of large fluctuations in nonequilibrium systems evolving according to Langevin's equations of motion. This technique is used to simulate the behavior of the bistable Maier-Stein system at noise intensities much lower than those previously possible. PMID- 11497654 TI - Noise properties of stochastic processes and entropy production. AB - Based on a Fokker-Planck description of external Ornstein-Uhlenbeck noise and cross-correlated noise processes driving a dynamical system we examine the interplay of the properties of noise processes and the dissipative characteristic of the dynamical system in the steady state entropy production and flux. Our analysis is illustrated with appropriate examples. PMID- 11497655 TI - Microscopic self-organization in networks. AB - We report our numerical studies on microscopic self-organization of a reaction system in three types of differently connected networks: a regular network, a small-world network, and a random network. Our simulation results show that the topology of the network has an important effect on the communication among reaction molecules, and plays an important role in microscopic self-organization. The correlation length among reacting molecules in a random or a small-world network is much shorter compared with that in a regular one. As a result, it is much easier to obtain microscopic self-organization in a small-world or a random network. We also observed a phase transition from a stochastic state to a synchronized state when we increased the randomness of a small-world network. PMID- 11497656 TI - Constant pressure molecular dynamics on a hypercylinder. AB - A Lagrangian formalism for variable-cell-shape molecular dynamics is derived from first principles. It is based on consideration of a crystal as arranged on the surface of hypercylinder in an extended coordinate frame. The artificial curvature along the additional degrees of freedom upsets the balance of forces acting on every atom in a periodically repeating cell. Since the distance between atoms is not the metric tensor the proposed method provides an essential simplification of the equations of motion compared to those of Parinello and Rahman approach. The Lagrangian of the system eliminates the cell orientation from the dynamics, thus avoiding symmetry-breaking effects and physically irrelevant cell rotation. The change from a fully flexible cell to an isotropically flexible cell is realized within the same computational framework. Simulations for the transformation of a model He lattice under isotropic applied pressure are used to illustrate the application of this method. PMID- 11497657 TI - Scaling theory and spreading dynamics in systems with one absorbing state derived from an equilibrium statistical model. AB - We show that for systems with one absorbing state, the widely assumed scaling properties of the survival probability and of the probability density of the size of activity avalanches cannot be true in the asymptotic limit. Trying to answer the question, what is the true asymptotic limit of these quantities, we study Domany-Kinzel probabilistic cellular automata using an equilibrium statistical mechanic model (ESM). We are able to express important quantities of the avalanche dynamics by correlation functions of the ESM. The application of scaling theory to the ESM allows for the derivation of the scaling properties of quantities of the avalanche dynamics in the form of infinite series. From these results we can obtain possible solutions for the apparent scaling problem, but cannot decide definitely which one is true. The most appealing solution, for which some evidence is given, states that there is a narrow range around the critical point in which, for example, the survival probability has the same power law behavior as on the critical point. Outside this narrow range, the usually assumed scaling should be approximately valid. PMID- 11497658 TI - Extremal optimization for graph partitioning. AB - Extremal optimization is a new general-purpose method for approximating solutions to hard optimization problems. We study the method in detail by way of the computationally hard (NP-hard) graph partitioning problem. We discuss the scaling behavior of extremal optimization, focusing on the convergence of the average run as a function of run time and system size. The method has a single free parameter, which we determine numerically and justify using a simple argument. On random graphs, our numerical results demonstrate that extremal optimization maintains consistent accuracy for increasing system sizes, with an approximation error decreasing over run time roughly as a power law t(-0.4). On geometrically structured graphs, the scaling of results from the average run suggests that these are far from optimal with large fluctuations between individual trials. But when only the best runs are considered, results consistent with theoretical arguments are recovered. PMID- 11497659 TI - Percolation threshold, Fisher exponent, and shortest path exponent for four and five dimensions. AB - We develop a method of constructing percolation clusters that allows us to build very large clusters using very little computer memory by limiting the maximum number of sites for which we maintain state information to a number of the order of the number of sites in the largest chemical shell of the cluster being created. The memory required to grow a cluster of mass s is of the order of s(straight theta) bytes where straight theta ranges from 0.4 for two-dimensional (2D) lattices to 0.5 for six (or higher)-dimensional lattices. We use this method to estimate d(min), the exponent relating the minimum path l to the Euclidean distance r, for 4D and 5D hypercubic lattices. Analyzing both site and bond percolation, we find d(min)=1.607+/-0.005 (4D) and d(min)=1.812+/-0.006 (5D). In order to determine d(min) to high precision, and without bias, it was necessary to first find precise values for the percolation threshold, p(c): p(c)=0.196889+/ 0.000003 (4D) and p(c)=0.14081+/-0.00001 (5D) for site and p(c)=0.160130+/ 0.000003 (4D) and p(c)=0.118174+/-0.000004 (5D) for bond percolation. We also calculate the Fisher exponent tau determined in the course of calculating the values of p(c): tau=2.313+/-0.003 (4D) and tau=2.412+/-0.004 (5D). PMID- 11497660 TI - Unstable decay and state selection. AB - The decay of unstable states when several metastable states are available for occupation is investigated using path-integral techniques. Specifically, a method is described that enables the probabilities with which the metastable states are occupied to be calculated by finding optimal paths, and fluctuations about them, in the weak-noise limit. The method is illustrated on a system described by two coupled Langevin equations, which are found in the study of instabilities in fluid dynamics and superconductivity. The problem involves a subtle interplay between nonlinearities and noise, and a naive approximation scheme that does not take this into account is shown to be unsatisfactory. The use of optimal paths is briefly reviewed and then applied to finding the conditional probability of ending up in one of the metastable states, having begun in the unstable state. There are several aspects of the calculation that distinguish it from most others involving optimal paths: (i) the paths do not begin and end on an attractor, and moreover, the final point is to a large extent arbitrary, (ii) the interplay between the fluctuations and the leading-order contribution are at the heart of the method, and (iii) the final result involves quantities that are not exponentially small in the noise strength. This final result, which gives the probability of a particular state being selected in terms of the parameters of the dynamics, is remarkably simple and agrees well with the results of numerical simulations. The method should be applicable to similar problems in a number of other areas, such as state selection in lasers, activationless chemical reactions, and population dynamics in fluctuating environments. PMID- 11497661 TI - Effects of noise coherence on stochastic resonance enhancement in a bithreshold system. AB - We identify a method for optimizing the stochastic resonance (SR) in a symmetric bithreshold device: by varying the coherence of the added noise series. To show SR enhancement via this method, we compare the performance of the system using noise sources with different coherence at normalized amplitude. The normalization of the noise amplitude is based on the mean threshold crossing rate of the Gaussian white noise, which is considered as the standard noise in SR studies, at optimal variance. The amplitude for optimal performance of the Gaussian white noise is determined using a signal-to-noise ratio (Q). The Q measure is also used to compare and examine the system performance for different noise cases. This measure is used because it is particularly sensitive to the effects of coherence on the quality of the output power spectrum. PMID- 11497662 TI - Random graphs with arbitrary degree distributions and their applications. AB - Recent work on the structure of social networks and the internet has focused attention on graphs with distributions of vertex degree that are significantly different from the Poisson degree distributions that have been widely studied in the past. In this paper we develop in detail the theory of random graphs with arbitrary degree distributions. In addition to simple undirected, unipartite graphs, we examine the properties of directed and bipartite graphs. Among other results, we derive exact expressions for the position of the phase transition at which a giant component first forms, the mean component size, the size of the giant component if there is one, the mean number of vertices a certain distance away from a randomly chosen vertex, and the average vertex-vertex distance within a graph. We apply our theory to some real-world graphs, including the world-wide web and collaboration graphs of scientists and Fortune 1000 company directors. We demonstrate that in some cases random graphs with appropriate distributions of vertex degree predict with surprising accuracy the behavior of the real world, while in others there is a measurable discrepancy between theory and reality, perhaps indicating the presence of additional social structure in the network that is not captured by the random graph. PMID- 11497663 TI - Relevance of percolation theory to the vulcanization transition. AB - The relationship between vulcanization and percolation is explored from the perspective of renormalized local field theory. We show to arbitrary order in perturbation theory that the vulcanization and percolation correlation functions are governed by the same Gell-Mann-Low renormalization-group equation. Hence, all scaling aspects of the vulcanization transition are reigned by the critical exponents of the percolation universality class. PMID- 11497664 TI - Adiabatic elimination and reduced probability distribution functions in spatially extended systems with a fluctuating control parameter. AB - We obtain the stationary probability distribution functions of the order parameter near onset for the one-dimensional real Ginzburg-Landau and Swift Hohenberg equations with a fluctuating control parameter. A perturbative expansion in the intensity of the fluctuations leads to a hierarchy of Fokker Planck equations for conditional probability distribution functions that relate components of the order parameter that evolve in different time scales. Successive integration leads to a Fokker-Planck equation for the slowest mode, which we solve analytically for the models studied. In all cases, the probability distribution function above onset is of the form P(A0) approximately A(delta)(0)e(-gammaA20), where A0 is the slow component of the order parameter and the values of delta and gamma depend explicitly on the intensity of the fluctuations. Knowledge of P(A0) allows the calculation of an effective bifurcation threshold and of the moments of A0 above threshold. PMID- 11497665 TI - Fractional non-Brownian motion and trapping-time distributions of grains in rice piles. AB - Non-Gaussian height fluctuations occurring on the fueling time scale of a slowly driven rice pile match those observed in some turbulent/critical phenomena, forming an anticorrelated random fractal process with Hurst exponent H=0.2. Inspired by this observation, the concept of fractional Brownian motion (FBM) is extended to treat stochastic processes with skewed increments. Simulations of this process for antipersistent motion have first return time distribution deviating from the t(-2+H) law for FBM. The first return time distribution of this fractional non-Brownian motion describes and quantitatively determines the trapping-time distribution of grains in rice piles upon incorporating a continuous representation of the additional height fluctuations that occur on the time scale between fueling events. PMID- 11497666 TI - Construction of an effective Hamiltonian for a three-dimensional Ising universality class. AB - The asymptotic and preasymptotic critical behavior in fluids, mixtures, and uniaxial magnets is believed to be described by an effective straight phi(4) scalar field theory with suitable, nonuniversal, coupling constants. The critical parameters as well as the extent of crossovers and corrections to the leading critical behavior in physical systems, crucially depends on the choice of these couplings. Here we propose a new method for deriving the effective scalar field theory appropriate to a microscopic model in this universality class. Use is made of the hierarchical reference theory, which implements the basic ideas of Wilson momentum space renormalization group to microscopic Hamiltonians. The effective low-energy field theory is then analyzed by the minimal subtraction scheme of Schloms and Dohm. We discuss the application of this method to the three dimensional Ising model and to the liquid-vapor phase transition. We make comparison with high-temperature expansion results and with experimental data for rare gas. PMID- 11497667 TI - Crossover between ordinary and normal transitions in the presence of a bulk field. AB - We investigate two-dimensional Ising films at the critical temperature T(c) and nonzero bulk magnetic field h using the density-matrix renormalization-group method. The crossover between ordinary (h(1)=0) and normal (h(1)=infinity) transitions corresponding to finite values of the surface fields h(1)=h(2), is studied. The structure and the solvation force f(solv) as a function of h, crucially depend on the value of h(1). Scaling functions for f(solv) and the longitudinal correlation length are given and discussed. PMID- 11497668 TI - Unconventional decay law for excited states in closed many-body systems. AB - We study the time evolution of an initially excited many-body state in a finite system of interacting Fermi particles in the situation when the interaction gives rise to the "chaotic" structure of compound states. This situation is generic for highly excited many-particle states in quantum systems such as heavy nuclei, complex atoms, quantum dots, spin systems, and quantum computers. For a strong interaction the leading term for the return probability W(t) has the form W(t) approximately exp(-Delta(2)(E)t(2)) with Delta(2)(E) as the variance of the strength function. The conventional exponential linear dependence W(t)=C exp( Gammat) formally arises for a very larger time. However, the prefactor C turns out to be exponentially large, thus resulting in a strong difference from the conventional estimate for W(t). PMID- 11497669 TI - Crossover parametric equation of state for Ising-like systems. AB - We present a parametric equation for the thermodynamic properties in the critical region of three-dimensional Ising-like systems which include fluids and fluid mixtures. The equation of state incorporates a crossover from singular Ising behavior asymptotically close to the critical point to classical (mean-field) behavior further away from the critical point, characterized by two physical crossover parameters: a coupling constant related to the strength and range of molecular interactions and a "cutoff" wave number for the critical fluctuations. In the asymptotic Ising limit, the crossover equation reproduces the most recent theoretical estimates for the universal ratios of the leading and correction-to scaling critical amplitudes. The equation has been tested by comparing it with recent experimental thermodynamic-property data for 3He near its vapor-liquid critical point. PMID- 11497670 TI - Symmetry breaking and phase coexistence in a driven diffusive two-channel system. AB - We consider classical hard-core particles moving on two parallel chains in the same direction. An interaction between the channels is included via the hopping rates. For a ring, the stationary state has a product form. For the case of coupling to two reservoirs, it is investigated analytically and numerically. In addition to the known one-channel phases, two new regions are found, in particular one, where the total density is fixed, but the filling of the individual chains changes back and forth, with a preference for strongly different densities. The corresponding probability distribution is determined and shown to have a universal form. The phase diagram and general aspects of the problem are discussed. PMID- 11497671 TI - Origin of quantum chaos for two particles interacting by short-range potentials. AB - We address the problem of two confined one-dimensional particles of arbitrary masses interacting by general short-range potentials. We study under what conditions quantum chaos emerges for the system by analyzing its spectrum statistics. We show that these conditions are directly connected with a specific feature of the underlying classical dynamics, namely, the ergodicity in the changes of the particles momenta. Quantum mechanically this prevents one from obtaining the exact wave function through the Bethe ansatz. Possible extensions for many-body systems are also discussed. PMID- 11497672 TI - Characteristics of a piecewise smooth area-preserving map. AB - We are reporting a study carried out in a system concatenated by two area preserving maps. The system can be viewed as a model of an electronic relaxation oscillator with over-voltage protection. We found that a border-collision bifurcation may interrupt a period-doubling bifurcation cascade, and that some special features, such as "quasicoexisting periodic orbits crossing border" as well as the transition between "quasitransience" and chaotic orbits, accompany the process. These features belong to the so-called "quasidissipative" properties. Here "quasitransience" denotes the behavior of iterations outside elliptic islands. They are "attracted" to the islands. As soon as it reaches the islands, the iteration follows the conservative regulations exactly. This induces a kind of escaping from strange sets. The scaling behavior of the escaping rate is obtained numerically. PMID- 11497673 TI - Effects of geometrical ray chaos on the electromagnetic eigenmodes of a gradient index optical cavity. AB - Electromagnetic analogies of quantum chaos are investigated in two-dimensional optical cavities which have reflective surfaces and contain a gradient refractive index medium. As the shape of the cavity is transformed continuously from a rectangle to a parallelogram, the geometrical ray paths undergo a transition from stable to chaotic dynamics. In the chaotic regime, the spectral statistics of the cavity are accurately described by random matrix theory. In addition, the electromagnetic mode spectrum of the cavity is modulated by both real and ghost periodic ray paths. These paths also "scar" the electric field intensity distributions of regular subsets of cavity eigenmodes. PMID- 11497674 TI - Clustering of noise-induced oscillations. AB - The subject of our study is clustering in a population of excitable systems driven by Gaussian white noise and with randomly distributed coupling strength. The cluster state is frequency-locked state in which all functional units run at the same noise-induced frequency. Cooperative dynamics of this regime is described in terms of effective synchronization and noise-induced coherence. PMID- 11497675 TI - Loss of coherence in a system of globally coupled maps. AB - We study the formation of symmetric (i.e., equally sized) or nearly symmetric clusters in an ensemble of globally coupled, identical chaotic maps. It is shown that the loss of synchronization for the coherent state and the emergence of subgroups of oscillators with synchronized behavior are two distinct processes, and that the type of behavior that arises after the loss of total synchronization depends sensitively on the dynamics of the individual map. For our system of globally coupled logistic maps, symmetric two-cluster formation is found to proceed through a periodic state associated with the stabilization either of an asynchronous period-2 cycle or of an asynchronous period-4 cycle. With further reduction of the coupling strength, each of the principal clustering states undergoes additional bifurcations leading to cycles of higher periodicity or to quasiperiodic and chaotic dynamics. If desynchronization of the coherent chaotic state occurs before the formation of stable clusters becomes possible, high dimensional chaotic motion is observed in the intermediate parameter interval. PMID- 11497676 TI - Phase synchronization of diffusively coupled Rossler oscillators with funnel attractors. AB - Recently, an antiphase phase-synchronized state in a system of diffusively coupled Rossler oscillators has been reported [Gang Hu et al., Phys. Rev. Lett. 85, 3377 (2000)]. In the current paper this antiphase state is explored in detail. Our interests are concentrated on the comparison with the normal in-phase phase-synchronized state for phase-coherent oscillators and the effect of the lattice size. Our main results are that (i) this antiphase synchronization is only for funnel Rossler attractors and cannot be observed in a system of coupled phase-coherent oscillators; (ii) it can be observed only for intermediate values of the lattice size while it disappears for quite low or large values of the lattice size; and (iii) it is different from the in-phase phase-synchronized state of phase-coherent oscillators in many respects. PMID- 11497677 TI - Domain coexistence in nonlinear optical pattern formation. AB - We report on domain coexistence of a variety of different modes in a two dimensional nonlinear optoelectronic model. The changes of stabilities of these modes are shown to give rise to new forms of spatial and spatiotemporal structures. The existence of these domain patterns explains some of the patterns recently observed in an optical system with a large array optoelectronic feedback circuit. PMID- 11497678 TI - Pattern formation on trees. AB - Networks having the geometry and the connectivity of trees are considered as the spatial support of spatiotemporal dynamical processes. A tree is characterized by two parameters: its ramification and its depth. The local dynamics at the nodes of a tree is described by a nonlinear map, giving rise to a coupled map lattice system. The coupling is expressed by a matrix whose eigenvectors constitute a basis on which spatial patterns on trees can be expressed by linear combination. The spectrum of eigenvalues of the coupling matrix exhibit a nonuniform distribution that manifests itself in the bifurcation structure of the spatially synchronized modes. These models may describe reaction-diffusion processes and several other phenomena occurring on heterogeneous media with hierarchical structure. PMID- 11497679 TI - Explosion of limit cycles and chaotic waves in a simple nonlinear chemical system. AB - We consider a simple model of an autocatalytic chemical reaction where a limit cycle rapidly increases to infinite period and amplitude, and disappears under variation of a parameter. We show that this bifurcation can be understood from seeing the system as a singular perturbation problem, and we find the bifurcation point by an asymptotic analysis. Scaling laws for period and amplitude are derived. The unphysical bifurcation to infinity disappears under generic modifications of the model, and for a simple example we show is replaced by a canard explosion, that is, a narrow parameter interval with an explosive growth of the amplitude. The bifurcation to infinity introduces a strong sensitivity that may result in chaotic dynamics if diffusion is added. We show that this behavior persists even if the kinetics is modified to preclude the bifurcation to infinity. PMID- 11497680 TI - Sporadic randomness: the transition from the stationary to the nonstationary condition. AB - We address the study of sporadic randomness by means of the Manneville map. We point out that the Manneville map is the generator of fluctuations yielding the Levy processes, and that these processes are currently regarded by some authors as statistical manifestations of a nonextensive form of thermodynamics. For this reason we study the sensitivity to initial conditions with the help of a nonextensive form of the Lyapunov coefficient. The purpose of this research is twofold. The former is to assess whether a finite diffusion coefficient might emerge from the nonextensive approach. This property, at first sight, seems to be plausible in the nonstationary case, where conventional Kolmogorov-Sinai analysis predicts a vanishing Lyapunov coefficient. The latter purpose is to confirm or reject conjectures about the nonextensive nature of Levy processes. We find that the adoption of a nonextensive approach does not serve any predictive purpose: It does not even signal a transition from a stationary to a nonstationary regime. These conclusions are reached by means of both numerical and analytical calculations that shed light on why the Levy processes do not imply any need to depart from the adoption of traditional complexity measures. PMID- 11497681 TI - Diffusion by extrinsic noise in a two-dimensional anisotropic web mapping. AB - Diffusion by an extrinsic noise is studied in a two-dimensional anisotropic web mapping where the intrinsic web diffusion is negligible, diffusion in one direction is dominant over the other, and the extrinsic noise enters in the dominant dimension only. It is found that the diffusion scaling is governed by the competition between the extrinsic noise and the intrinsic rotation. If the extrinsic noise is weaker than the intrinsic rotation, diffusion scales as lK(1/2) in the dominant direction and as lK(3/2) in the nondominant direction, where l is the extrinsic noise strength and K is the intrinsic perturbation parameter. If the extrinsic noise is stronger, diffusion behaves as l(2)/2 in the dominant direction and as K(2)/4 in the nondominant direction. Diffusion in the nondominant direction can be important if the equilibrium system is translationally invariant in the dominant direction. PMID- 11497682 TI - Quantum-classical correspondence in polygonal billiards. AB - We show that wave functions in planar rational polygonal billiards (all angles rationally related to pi) can be expanded in a basis of quasistationary and spatially regular states. Unlike the energy eigenstates, these states are directly related to the classical invariant surfaces in the semiclassical limit. This is illustrated for the barrier billiard. We expect that these states are also present in integrable billiards with point scatterers or magnetic-flux lines. PMID- 11497683 TI - Extracting dynamical structure from unstable periodic orbits. AB - The topological recurrence algorithm provides a fast and robust method for detecting the presence of unstable periodic orbits (UPO's) in short, noisy experimental data files. We present here a technique for improving this method by using a matrix fitting algorithm to extract dynamical information about the system from these UPO's. This method greatly increases the sensitivity of the algorithm, and also provides a method for identifying false positive results. PMID- 11497684 TI - Detecting unstable periodic orbits in chaotic continuous-time dynamical systems. AB - We extend the recently developed method for detecting unstable periodic points of chaotic time-discrete dynamical systems to find unstable periodic orbits in time continuous systems, given by a set of ordinary differential equations. This is achieved by the reduction of the continuous flow to a Poincare map which is then searched for periodic points. The algorithm has global convergence properties and needs no a priori knowledge of the system. It works well for both dissipative and Hamiltonian dynamical systems which is demonstrated by exploring the Lorenz system and the hydrogen atom in a strong magnetic field. The advantages and general features of the approach are discussed in detail. PMID- 11497685 TI - Replacement manifolds: a method to uniformize semiclassical wave functions. AB - We present a semiclassical technique that relies on replacing complicated classical manifold structure with simpler manifolds, which are then evaluated by the usual semiclassical rules. Under circumstances where the original manifold structure gives poor or useless results semiclassically the replacement manifolds can yield remarkable accuracy. We give several working examples to illustrate the theory presented here. PMID- 11497686 TI - Chaos in a Jahn-Teller molecule. AB - The Jahn-Teller system [see text] has a particular degeneracy, where the vibronic potential has an elliptical minimum. In the general case where the ellipse does not reduce to a circle, the classical motion in the potential is chaotic, tending to trapping near one of the extrema of the ellipse. In the quantum problem, the motion consists of correlated tunneling from one extremum to the opposite, leading to an average angular momentum reminiscent of that of the better known [see text] dynamic Jahn-Teller system. PMID- 11497687 TI - Quantum-classical correspondence for the equilibrium distributions of two interacting spins. AB - We consider the quantum and classical Liouville dynamics of a nonintegrable model of two coupled spins. Initially localized quantum states spread exponentially to the system size when the classical dynamics are chaotic. The long-time behavior of the quantum probability distributions and, in particular, the parameter dependent rates of relaxation to the equilibrium state are surprisingly well approximated by the classical Liouville mechanics even for small quantum numbers. As the accessible classical phase space becomes predominantly chaotic, the classical and quantum probability equilibrium configurations approach the microcanonical distribution, although the quantum equilibrium distributions exhibit characteristic "minimum" fluctuations away from the microcanonical state. The magnitudes of the quantum-classical differences arising from the equilibrium quantum fluctuations are studied for both pure and mixed (dynamically entangled) quantum states. In both cases the standard deviation of these fluctuations decreases as (h/J)(1/2), where J is a measure of the system size. In conclusion, under a variety of conditions the differences between quantum and classical Liouville mechanics are shown to become vanishingly small in the classical limit (J/h-->infinity) of a nondissipative model endowed with only a few degrees of freedom. PMID- 11497688 TI - Local and global control of high-period unstable orbits in reversible maps. AB - We study the nonlinear dynamics of a complex system, described by a two dimensional reversible map. The phase space of this map exhibits elements typical of Hamiltonian systems (stability islands) as well as of dissipative systems (attractor). Due to the interaction between the stability islands and the attractor, the transition to chaos in this system occurs through the collapse of the stability island and stochastization of the limiting-cycles orbits. We show how to apply the method of discrete parametric control to stabilize unstable high period orbits. To achieve highly efficient control we introduce the concepts of local and global control. These concepts are useful in situations where there are "dangerous" points on the target orbit, i.e., the points where the probability of breakdown of control is high. As a result, the dangerous points turn out to be much more sensitive to external noise than other points on the orbit, and only the dangerous points determine how effective the control is. PMID- 11497689 TI - Spatiotemporal dynamics near a supercritical Turing-Hopf bifurcation in a two dimensional reaction-diffusion system. AB - Pattern formation in semiconductor heterostructures is studied on the basis of a spatially two-dimensional model of reaction-diffusion type. In particular, we investigate the neighborhood of a codimension-two Turing-Hopf instability by analytical methods. Amplitude equations are derived which predict the absence of mixed modes but extended ranges of bistability between homogeneous oscillatory states and hexagonal Turing patterns. Our results are confirmed by numerical simulations. The features are not confined to a neighborhood of the bifurcation point so that the conclusions of the weakly nonlinear analysis explain the observations in large portions of the parameter space at least qualitatively PMID- 11497690 TI - Positive Lyapunov exponents calculated from time series of strange nonchaotic attractors. AB - Time-series methods for estimating Lyapunov exponents may give a positive exponent when they are applied to the time series of strange nonchaotic systems. Strange nonchaotic systems are characterized by expanding and contracting regions in phase space that result in repeatedly expanding or contracting trajectories. Using time-series methods, the maximum time-series Lyapunov exponent is calculated as an average of the locally most expanding exponents that characterize the divergence of nearby trajectories following a reconstructed attractor over time. A positive exponent is reported by time-series methods for trajectories in an expanding region. While in a converging region, the most expanding dynamics are related to the quasiperiodic driving force. Statistically, a zero exponent related to the quasiperiodic force is obtained through time series methods within converging regions. As a result, the calculated maximum Lyapunov exponent is positive. PMID- 11497691 TI - Detecting functional relationships between simultaneous time series. AB - We describe a method to characterize the predictability and functionality between two simultaneously generated time series. This nonlinear method requires minimal assumptions and can be applied to data measured either from coupled systems or from different positions on a spatially extended system. This analysis generates a function statistic, Theta(c(0)), that quantifies the level of predictability between two time series. We illustrate the utility of this procedure by presenting results from a computer simulation and two experimental systems. PMID- 11497692 TI - Stochastic aspects of one-dimensional discrete dynamical systems: Benford's law. AB - Benford's law owes its discovery to the "Grubby Pages Hypothesis," a 19th century observation made by Simon Newcomb that the beginning pages of logarithm books were grubbier than the last few pages, implying that scientists referenced the values toward the front of the books more frequently. If a data set satisfies Benford's law, then it's significant digits will have a logarithmic distribution, which favors smaller significant digits. In this article we demonstrate two ways of creating discrete one-dimensional dynamical systems that satisfy Benford's law. We also develop a numerical simulation methodology that we use to study dynamical systems when analytical results are not readily available. PMID- 11497693 TI - Translational motion of two interacting bubbles in a strong acoustic field. AB - Using the Lagrangian formalism, equations of radial and translational motions of two coupled spherical gas bubbles have been derived up to terms of third order in the inverse distance between the bubbles. The equations of radial pulsations were then modified, for the purpose of allowing for effects of liquid compressibility, using Keller-Miksis' approach, and the equations of translation were added by viscous forces in the form of the Levich drag. This model was then used in a numerical investigation of the translational motion of two small, driven well below resonance, bubbles in strong acoustic fields with pressure amplitudes exceeding 1 bar. It has been found that, if the forcing is strong enough, the bubbles form a bound pair with a steady spacing rather than collide and coalesce, as classical Bjerknes theory predicts. Moreover, the viscous forces cause skewness in the system, which results in self-propulsion of the bubble pair. The latter travels as a unit along the center line in a direction that is determined by the ratio of the initial bubble radii. The results obtained are of immediate interest for understanding and modeling collective bubble phenomena in strong fields, such as acoustic cavitation streamers. PMID- 11497694 TI - Effect of large-scale intermittency and mean shear on scaling-range exponents in a turbulent jet. AB - The present study investigates the combined impact of the intermittency associated with the turbulent-nonturbulent interface and the mean shear rate in an axisymmetric jet on the structure of turbulence in the scaling range, where the spectrum exhibits a power-law behavior. Second-order structure functions, autocorrelations of the dissipation rate, and spectra of both the longitudinal velocity fluctuation and the passive temperature fluctuation are measured at a distance of 40 diameter downstream from the nozzle exit. All the scaling range exponents are influenced by the large-scale intermittency and the mean shear. The scalar fluctuation is much more sensitive to the variation in large-scale intermittency than the velocity fluctuation. PMID- 11497695 TI - Thermodynamics of fluid turbulence: a unified approach to the maximum transport properties. AB - Dissipative properties of various kinds of turbulent phenomena are investigated. Two expressions are derived for the rate of entropy increase due to thermal and viscous dissipation by turbulence, and for the rate of entropy increase in the surrounding system; both rates must be equal when the fluid system is in a steady state. Possibility is shown with these expressions that the steady-state properties of several different types of turbulent phenomena (Benard-type thermal convection, turbulent shear flow, and the general circulation of the atmosphere and ocean) exhibit a unique state in which the rate of entropy increase in the surrounding system by the turbulent dissipation is at a maximum. The result suggests that the turbulent fluid system tends to be in a steady state with a distribution of eddies that produce the maximum rate of entropy increase in the nonequilibrium surroundings. PMID- 11497696 TI - Mean-field theory for a passive scalar advected by a turbulent velocity field with a random renewal time. AB - Mean-field theory for turbulent transport of a passive scalar (e.g., particles and gases) is discussed. Equations for the mean number density of particles advected by a random velocity field, with a finite correlation time, are derived. Mean-field equations for a passive scalar comprise spatial derivatives of high orders due to the nonlocal nature of passive scalar transport in a random velocity field with a finite correlation time. A turbulent velocity field with a random renewal time is considered. This model is more realistic than that with a constant renewal time used by Elperin et al. [Phys. Rev. E 61, 2617 (2000)], and employs two characteristic times: the correlation time of a random velocity field tau(c), and a mean renewal time tau. It is demonstrated that the turbulent diffusion coefficient is determined by the minimum of the times tau(c) and tau. The mean-field equation for a passive scalar was derived for different ratios of tau/tau(c). The important role of the statistics of the field of Lagrangian trajectories in turbulent transport of a passive scalar, in a random velocity field with a finite correlation time, is demonstrated. It is shown that in the case tau(c)< or =d(c) approximately 2.2. For Kolmogorov's solution, both renormalized viscosity and resistivity are positive for the whole range of parameters. Various cascade rates and Kolmogorov's constant for MHD turbulence have been calculated by solving the flux equation to first order in the perturbation series. We find that the magnetic energy cascades forward. The Kolmogorov constant for d=3 does not vary significantly with r(A) and is found to be close to the constant for fluid turbulence. PMID- 11497698 TI - Large negative velocity gradients in Burgers turbulence. AB - We consider one-dimensional Burgers equation driven by large-scale white-in-time random force. The tails of the velocity gradients probability distribution function (PDF) are analyzed by saddle point approximation in the path integral describing the velocity statistics. The structure of the saddle-point (instanton), that is, the velocity field configuration realizing the maximum of probability, is studied numerically in details. The numerical results allow us to find analytical solution for the long-time part of the instanton. Its careful analysis confirms the result of Balkovsky et al. [Phys. Rev. Lett. 78, 1452 (1997)] based on short-time estimations that the left tail of PDF has the form ln P(u(x))infinity-/u(x)/(3/2). PMID- 11497699 TI - Inertial effects on reactive particles advected by turbulence. AB - We study the problem of the advection of passive particles with inertia in a two dimensional, synthetic, and stationary turbulent flow. The asymptotic analytical result and numerical simulations show the importance of inertial bias in collecting the particles preferentially in certain regions of the flow, depending on their density relative to that of the flow. We also study how these aggregates are affected when a simple chemical reaction mechanism is introduced through a Eulerian scheme. We find that inertia can be responsible for maintaining a stationary concentration pattern even under nonfavorable reactive conditions or destroying it under favorable ones. PMID- 11497700 TI - Gap size effects for the Kelvin-Helmholtz instability in a Hele-Shaw cell. AB - We report experimental results for the Kelvin-Helmholtz instability between two immiscible fluids in parallel flow in a Hele-Shaw cell. We focus our interest on the influence of the gap size between the walls on the instability characteristics. Experimental results show that the instability threshold, the critical wavelength, the phase velocity, and the spatial growth rate depend on this gap size. These results are compared to both the previous two-dimensional analysis of Gondret and Rabaud [Phys. Fluids 9, 3267 (1997)] and the three dimensional analysis recently derived by Plouraboue and Hinch [Phys. Fluids (to be published)], showing that the agreement is still not complete especially when gap size increases. PMID- 11497701 TI - Kinetic theory of point vortices: diffusion coefficient and systematic drift. AB - We develop a kinetic theory for point vortices in two-dimensional hydrodynamics. Using standard projection operator techniques, we derive a Fokker-Planck equation describing the relaxation of a "test" vortex in a bath of "field" vortices at statistical equilibrium. The relaxation is due to the combined effect of a diffusion and a drift. The drift is shown to be responsible for the organization of point vortices at negative temperatures. A description that goes beyond the thermal bath approximation is attempted. A new kinetic equation is obtained which respects all conservation laws of the point vortex system and satisfies a H theorem. Close to equilibrium, this equation reduces to the ordinary Fokker Planck equation. PMID- 11497702 TI - Near-infrared emissions in single-bubble and multibubble sonoluminescence. AB - Comparisons of light emissions from multibubble and single-bubble sonoluminescence in the near-infrared band extending from 800 to 1050 nm have been investigated. In argon-water mixtures, single-bubble and multibubble spectra are similar in appearance. In sodium chloride-water mixtures, the multibubble spectrum shows evidence of the 3d-3p sodium emission line, whereas the single bubble spectrum shows no such emission. For single bubbles, the near-infrared emissions change linearly with driving pressure. No evidence of near-infrared emissions are observed below the visible luminescence threshold. PMID- 11497703 TI - Quantum mechanical representation of acoustic streaming and acoustic radiation pressure. AB - We discuss acoustic streaming and acoustic radiation pressure from the viewpoint of energy and momentum of acoustic waves, using a quantum mechanical representation of acoustic waves. We represent the energy epsilon and momentum mu of acoustic waves as epsilon=n(p)homega and mu=n(p)hk; here n(p) is the phonon density, omega is the frequency, k is the wave number, and h is Planck's constant. It is easy to derive the momentum of acoustic waves as mu=epsilon/c (c is the sound velocity). Therefore, we can represent the acoustic streaming and acoustic radiation pressure in terms of the momentum. PMID- 11497704 TI - Asymptotic analysis and renormalized perturbation theory of the non-Hermitian dynamics of an inviscid vortex. AB - An analysis of the non-Hermitian fluid systems described by the Rayleigh equation in an unbounded domain has been carried out in the regime of large wave numbers. The evolution of a special class of localized vorticities is also discussed. Asymptotic and perturbative approaches lead to the same final result. In the limit considered, the system is stable. The perturbation analysis reveals interesting pathologies of the non-Hermitian systems. Under specific conditions, the expansion is found to show secular growth. A discussion about the mechanism of insurgence of such singular behavior is presented. It is also shown that the divergent expansion is renormalizable by means of the renormalization group method-the renormalized results are in complete conformity with the asymptotic solutions. PMID- 11497705 TI - Apparent secondary-electron emission coefficient and the voltage-current characteristics of argon glow discharges. AB - The accuracy of secondary-electron emission coefficients, that are used as input data of discharge models, seriously influences the calculated discharge characteristics. As it is very difficult to consider all possible electron emission processes of a cold cathode separately, in most of the recent models an apparent secondary coefficient gamma is applied, which is often assumed to be constant, even for a wide range of discharge conditions. In contrast with this common assumption, the present calculations-based on a heavy-particle hybrid model-show that in abnormal glow discharges gamma varies considerably with changing discharge conditions: a factor of 3 change of gamma has been found in the range of reduced current densities (0.04 mA cm(-2) Torr(-2)< or =j/p(2)< or =4 mA cm(-2) Torr(-2)) covered in this study. The present simulations also confirm that ionization by heavy particles plays a significant role in the ion production at the abnormal cathode fall. Moreover, it is shown, that the fast heavy particles reflected from the cathode surface play the dominant role in the gas heating. PMID- 11497706 TI - Time-resolved imaging of spatiotemporal patterns in a one-dimensional dielectric barrier discharge system. AB - A makeshift streak camera produces time-resolved images of filament patterns in a one-dimensional dielectric-barrier discharge system. The images reveal a variety of spatial and temporal structures, including temporally distinct discharge stages that occur at specific phases of the 12.5 kHz driving oscillation and spatially periodic filament arrays that are generated during the discharge stages. The data answer recent questions regarding time-averaged images of a similar system. Most notably, all but the simplest time-averaged images are due to the integration of multiple filament patterns generated in different discharge stages. Disordered states of the system are also examined. Some of these are spatially disordered but display temporal structure; others appear to lack both spatial and temporal order. Possibilities are suggested for investigations of surface-charge spreading and pattern stability in similar systems. PMID- 11497707 TI - Interaction of a rodlike charged macroparticle with a flowing plasma. AB - The formation of the wake potential is demonstrated for a rodlike particle in a plasma with an ion flow. The cases of various charge distributions on the rod are studied, including those contributing to the dipole momentum, and different angles between the direction of the ion flow and the rod are considered. The possibilities of the formation of lattices composed of cylindrical macroparticles in a plasma are discussed. PMID- 11497708 TI - Propagation of a randomized 600-ps laser beam in a helium gas jet over long scale lengths. AB - The propagation and interaction of a randomized 600-ps laser with a helium gas jet were studied experimentally for laser intensities of 10(14) W/cm(2). Such a study is of interest for the indirectly driven inertial confinement fusion scheme, where a randomized laser beam propagates into a gas-filled cavity over a distance of a few millimeters. The dynamics of ionization was studied using time resolved interferometry. Maps of electronic density n(e)(z,t) were retrieved from time resolved interferograms. The plasma temperature was studied using Thomson scattering. The results show that the laser diffracts while propagating, leading to a decrease in laser intensity and causing ionization to occur later in time. An ionization front, moving at a velocity of about v(f) approximately 2.8 x 10(6) m/s, was observed. Beam diffraction also causes a nonhomogeneous heating of the plasma: the entrance of the plasma is hotter than the exit. A one-dimensional model was used to fit the results. It takes into account collisional ionization and heating by inverse bremsstrahlung. The model shows very good agreement with the experiment. PMID- 11497709 TI - Harmonics generation in electron-ion collisions in a short laser pulse. AB - Anomalously high generation efficiency of coherent higher field harmonics in collisions between oppositely charged particles in the field of femtosecond lasers is predicted. This is based on rigorous numerical solutions of a quantum kinetic equation for dense laser plasmas that overcomes limitations of previous investigations. PMID- 11497710 TI - Lorentz force effects on the electron energy distribution in inductively coupled plasmas. AB - Depletion of the electron energy distribution function by slow electrons in the skin layer has been observed in experiment in a cylindrical inductive discharge with a flat coil at low frequency and low gas pressure. The origin of the effect lies in the dependence of the ponderomotive force (caused by the rf magnetic field) on the electron thermal motion under conditions of the anomalous skin effect. Analysis of the electron energy distribution based on the existence of adiabatic invariants for collisionless electron motion at low frequencies reveals enhanced anisotropy and time dependence of the electron distribution function due to strong rf magnetic fields and a polarization electrostatic potential at twice the driving frequency. The electron energy distributions calculated in the skin layer using experimentally measured electromagnetic fields and rf and dc potential profiles are in reasonable agreement with the experimental electron distributions. PMID- 11497711 TI - Long-time discrete particle effects versus kinetic theory in the self-consistent single-wave model. AB - The influence of the finite number N of particles coupled to a monochromatic wave in a collisionless plasma is investigated. For growth as well as damping of the wave, discrete particle numerical simulations show an N-dependent long time behavior resulting from the dynamics of individual particles. This behavior differs from the one due to the numerical errors incurred by Vlasov approaches. Trapping oscillations are crucial to long time dynamics, as the wave oscillations are controlled by the particle distribution inhomogeneities and the pulsating separatrix crossings drive the relaxation towards thermal equilibrium. PMID- 11497712 TI - Transition from the C3-dominated discharge to the sooting plasma. AB - Mass spectrometry and photoemission spectroscopy of a graphite hollow cathode source identify the parameters of the transition from the C3-dominated discharge to the sooting plasma. The transition is a function of the shape and profile of a special cusp magnetic field B(z)(r, theta), the geometry of the source, the discharge current, and pressure. Characteristic atomic and molecular emission lines and bands in the C3 discharge transform into broad bands emitted by the excited soot. We identify four prominent emission bands between 300-400 nm to be the hallmark of the sooting plasma. PMID- 11497713 TI - Transport properties in a two-temperature plasma: theory and application. AB - An alternate derivation of transport properties in a two-temperature plasma has been performed. Indeed, recent works have shown that the simplified theory of transport properties out of thermal equilibrium introduced by Devoto and then Bonnefoi, very often used in two-temperature modeling, is questionable and particularly does not work when calculating the combined diffusion coefficients of Murphy. Thus, in this paper, transport properties are derived without Bonnefoi's assumptions in a nonreactive two-temperature plasma, assuming chemical equilibrium is achieved. The electron kinetic temperature T(e) is supposed to be different from that of heavy species T(h). Only elastic processes are considered in a collision-dominated plasma. The resolution of Boltzmann's equation, thanks to the Chapman-Enskog method, is used to calculate transport coefficients from sets of linear equations. The solution of these systems allows transport coefficients to be written as linear combinations of collision integrals, which take into account the interaction potential for a collision between two particles. These linear combinations are derived by extending the definition and the calculation of bracket integrals introduced by Chapman et al. to the thermal nonequilibrium case. The obtained results are rigorously the same as those of Hirschfelder et al. at thermal equilibrium. The derivation of diffusion velocity and heat flux shows the contribution of a new gradient, that of the temperature ratio straight theta=T(e)/T(h). An application is presented for a two-temperature argon plasma. First, it is shown that the two-temperature linear combinations of collision integrals are drastically modified with respect to equilibrium. Secondly, the two-temperature simplified theory of transport coefficients of Devoto and Bonnefoi underestimates the electron thermal conductivity with respect to the accurate value at T(e)=20 000 K. Lastly, contrary to the simplified theory of transport coefficients, the diffusion coefficients satisfy the symmetry conditions. An example is given at T(e)=6000 K for different values of straight theta for the diffusion coefficient between electrons and heavy species D(e-Ar) as well as for that between argon atoms and argon ions D(Ar-Ar+). PMID- 11497714 TI - Analytic models of high-temperature hohlraums. AB - A unified set of high-temperature-hohlraum models has been developed. For a simple hohlraum, P(S)=[A(S)+(1 alpha(W))A(W)+A(H)]sigmaT(4)(R)+(4Vsigma/c)(dT(4)(R)/dt), where P(S) is the total power radiated by the source, A(S) is the source area, A(W) is the area of the cavity wall excluding the source and holes in the wall, A(H) is the area of the holes, sigma is the Stefan-Boltzmann constant, T(R) is the radiation brightness temperature, V is the hohlraum volume, and c is the speed of light. The wall albedo alpha(W) identical with(T(W)/T(R))(4) where T(W) is the brightness temperature of area A(W). The net power radiated by the source P(N)=P(S) A(S)sigmaT(4)(R), which suggests that for laser-driven hohlraums the conversion efficiency eta(CE) be defined as P(N)/P(Laser). The characteristic time required to change T(4)(R) in response to a change in P(N) is 4V/c[(1-alpha(W))A(W)+A(H)]. Using this model, T(R), alpha(W), and eta(CE) can be expressed in terms of quantities directly measurable in a hohlraum experiment. For a steady-state hohlraum that encloses a convex capsule, P(N)=[(1-alpha(W))A(W)+A(H)+[(1 alpha(C))A(C)(A(S)+alpha(W)A(W))/A(T)]]sigmaT(4)(RC), where alpha(C) is the capsule albedo, A(C) is the capsule area, A(T) identical with(A(S)+A(W)+A(H)), and T(RC) is the brightness temperature of the radiation that drives the capsule. According to this relation, the capsule-coupling efficiency of the baseline National Ignition Facility hohlraum is 15-23 % higher than predicted by previous analytic expressions. A model of a hohlraum that encloses a z pinch is also presented. PMID- 11497715 TI - Absorption of femtosecond laser pulses in interaction with solid targets. AB - We have studied the effects of the plasma density scale length on the absorption mechanism of the femtosecond (fs) laser pulses interacting with solid targets. Experiments and particle-in-cell (PIC) simulations demonstrate that the vacuum heating is the main absorption in the plasma in the interaction of fs laser pulses with solid targets when no prepulses are applied. The energy spectrum of hot electrons ejected out of or injected into the plasma show a bitemperature distribution. While the first temperature of the two groups of hot electrons can be attributed to the "pull-and-push" exertion of the laser field, the second temperature refers to the electrons accelerated by the static part (in front of the target) and the oscillating part (in the plasma layer) of the laser-induced electric field, respectively. PIC simulations also show that with an appropriate density scale length, the femtosecond laser energy can be absorbed locally through different mechanisms. PMID- 11497716 TI - Motion of SF+x (x=1-3,5) and ion conversion in SF6-N2 mixtures. AB - This paper reports on the mass-analyzed measurement of the drift velocity of the positive SF6 daughter ions, SF+x (x=1-3,5) in the mixture of SF6 with N2 for SF6 concentrations of 10% and 50%. A double mass spectrometer-drift tube was used for these measurements. The density-normalized electric field intensity E/N, was varied from 30 to 360 Td (1 Td=10(-17) V cm(2)). We have found an increasingly monotonic dependence of the drift velocity with E/N, and an inverse dependence of the above parameter with the decrease of the SF6 content in the mixture. A test of Blanc's law for the zero-field reduced mobilities of the above ions shows a fair qualitative agreement. No drift velocities of these ionic species could be measured in pure N2, since we found that these ions reacted strongly with the nitrogen molecules. The cases for SF+3 and SF+5 are discussed. A similar behavior was observed for the nitrogen ions drifting in pure SF6. PMID- 11497717 TI - Nyquist method for Wigner-Poisson quantum plasmas. AB - By means of the Nyquist method, we investigate the linear stability of electrostatic waves in homogeneous equilibria of quantum plasmas described by the Wigner-Poisson system. We show that, unlike the classical Vlasov-Poisson system, the Wigner-Poisson case does not necessarily possess a Penrose functional determining its linear stability properties. The Nyquist method is then applied to a two-stream distribution, for which we obtain an exact, necessary and sufficient condition for linear stability, as well as to a bump-in-tail equilibrium. PMID- 11497718 TI - Nonlinear collisional absorption in dense laser plasmas. AB - Collisional absorption of dense, fully ionized plasmas in strong laser fields is investigated starting from a quantum kinetic equation with non-Markovian and field-dependent collision integrals in dynamically screened Born approximation. This allows to find rather general balance equations for the energy and the current. For high-frequency laser fields, quantum statistical expressions for the electrical current density and the cycle-averaged electron-ion collision frequency in terms of the Lindhard dielectric function are derived. The expressions are valid for arbitrary field strength assuming the nonrelativistic case. Numerical results are presented to discuss these quantities as a function of the applied laser field and for different plasma parameters. In particular, nonlinear phenomena such as higher harmonics generation and multiphoton emission and absorption in electron-ion collisions are considered. The significance to include quantum effects is demonstrated comparing our results for the collision frequency with previous results obtained from classical theories. PMID- 11497719 TI - Broadband optical gain via interference in the free electron laser: principles and proposed realizations. AB - We propose experimentally simplified schemes of an optically dispersive interface region between two coupled free electron lasers (FELs), aimed at achieving a much broader gain bandwidth than in a conventional FEL or a conventional optical klystron composed of two separated FELs. The proposed schemes can universally enhance the gain of FELs, regardless of their design, when operated in the short pulsed regime. PMID- 11497720 TI - Local energy exchange in a storage-ring free-electron laser. AB - In this paper a theoretical model is presented, which is based on a pass to pass analysis of the localized interaction between a short laser pulse with a wider electron distribution. It can be applied to a large class of physical phenomena and, in particular, to the case of a storage-ring free-electron laser (FEL). Numerical results are confirmed by experimental measurements done on the ACO and Super-ACO FELs. PMID- 11497721 TI - Invariants for time-dependent Hamiltonian systems. AB - An exact invariant is derived for n-degree-of-freedom Hamiltonian systems with general time-dependent potentials. The invariant is worked out in two equivalent ways. In the first approach, we define a special Ansatz for the invariant and determine its time-dependent coefficients. In the second approach, we perform a two-step canonical transformation of the initially time-dependent Hamiltonian to a time-independent one. The invariant is found to contain a function of time f(2)(t), defined as a solution of a linear third-order differential equation whose coefficients depend in general on the explicitly known configuration space trajectory that follows from the system's time evolution. It is shown that the invariant can be interpreted as the time integral of an energy balance equation. Our result is applied to a one-dimensional, time-dependent, damped non-linear oscillator, and to a three-dimensional system of Coulomb-interacting particles that are confined in a time-dependent quadratic external potential. We finally show that our results can be used to assess the accuracy of numerical simulations of time-dependent Hamiltonian systems. PMID- 11497722 TI - Apparent superluminal propagation of a laser pulse in a dispersive medium. AB - The distortion of a laser pulse propagating in a dispersive gain/absorptive medium is analyzed. The relationship between the distortion of the pulse and superluminal propagation is discussed. We present an analytical approach based on the laser envelope equation that is readily applicable to arbitrary input pulse shapes. This analysis is used to interpret recent experiments that claim to have observed distortionless superluminal laser pulse propagation. PMID- 11497723 TI - Microwave-induced control of free-electron-laser radiation. AB - The dynamical response of a relativistic bunch of electrons injected in a planar magnetic undulator and interacting with a counterpropagating electromagnetic wave is studied. We demonstrate a resonance condition for which the free-electron laser (FEL) dynamics is strongly influenced by the presence of the external field. It opens up the possibility of control of short wavelength FEL emission characteristics by changing the parameters of the microwave field without requiring change in the undulator's geometry or configuration. Numerical examples, assuming realistic parameter values analogous to those of the TTF-FEL, currently under development at DESY, are given for possible control of the amplitude or polarization of the emitted radiation. PMID- 11497724 TI - Discrete vortex solitons. AB - Localized states in the discrete two-dimensional (2D) nonlinear Schrodinger equation is found: vortex solitons with an integer vorticity S. While Hamiltonian lattices do not conserve angular momentum or the topological invariant related to it, we demonstrate that the soliton's vorticity may be conserved as a dynamical invariant. Linear stability analysis and direct simulations concur in showing that fundamental vortex solitons, with S=1, are stable if the intersite coupling C is smaller than some critical value C((1))(cr). At C>C((1))(cr), an instability sets in through a quartet of complex eigenvalues appearing in the linearized equations. Direct simulations reveal that an unstable vortex soliton with S=1 first splits into two usual solitons with S=0 (in accordance with the prediction of the linear analysis), but then an instability-induced spontaneous symmetry breaking takes place: one of the secondary solitons with S=0 decays into radiation, while the other one survives. We demonstrate that the usual (S=0) 2D solitons in the model become unstable, at C>C((0))(cr) approximately 2.46C((1))(cr), in a different way, via a pair of imaginary eigenvalues omega which bifurcate into instability through omega=0. Except for the lower-energy S=1 solitons that are centered on a site, we also construct ones which are centered between lattice sites which, however, have higher energy than the former. Vortex solitons with S=2 are found too, but they are always unstable. Solitons with S=1 and S=0 can form stable bound states. PMID- 11497725 TI - Iwasawa effects in multilayer optics. AB - There are many two-by-two matrices in layer optics. It is shown that they can be formulated in terms of a three-parameter group whose algebraic property is the same as the group of Lorentz transformations in a space with two spacelike and one timelike dimensions, or the Sp(2) group which is a standard theoretical tool in optics. Among the interesting mathematical properties of this group, the Iwasawa decomposition drastically simplifies the matrix algebra under certain conditions, and leads to a concise expression for the S matrix for transmitted and reflected waves. It is shown that the Iwasawa effect can be observed in multilayer optics, and a sample calculation of the S matrix is given. PMID- 11497726 TI - Fields of optical waveguides as waves in free space. AB - It is shown by using the scalar diffraction theory and the method of images that the arbitrary field confined by the optical waveguide can be generated in free space by the appropriate light source. The correspondence between the guided and free-space waves is illustrated using several particular fields, such as the diffraction-free, self-imaging, ultra-short, solitonlike, partially coherent waves and laser fractals. In opposition to the eigenmode theory of waveguides, the field at the guide entrance can satisfy neither the guide wave-equation nor the boundary conditions. PMID- 11497727 TI - Collisions between spatiotemporal solitons of different dimensionality in a planar waveguide. AB - A (2+1)-dimensional nonlinear Schrodinger equation including third-order dispersion is a natural model of a waveguide, in which strong temporal dispersion is induced by a grating in order to make the existence of two-dimensional spatiotemporal solitons possible. By means of analytical and numerical methods, we demonstrate that this model may support, simultaneously, stable dark quasi-one dimensional (stripe) solitons and two-dimensional elevation solitons ("antidark solitons") in the form of weakly localized "lumps." The spatial position of lumps can be controlled by passing stripe dark solitons through them in an arbitrary direction. To substantiate this mechanism, we analytically calculate a position shift generated by a headon collision between the stripe and lump. The obtained results are in good agreement with direct numerical simulations. PMID- 11497728 TI - Dielectrophoretic manipulation of macromolecules: the electric field. AB - The use of dielectrophoresis is fast becoming a proven technique for manipulating particles and macromolecules in microfluidic systems. Here an analytic solution for the gradient in the electric field strength, delta (E . E) [corrected] produced by a two-dimensional array of parallel electodes is derived using the method of Green's functions. The boundary condition for the potential between electrodes is estimated by using a linear approximation. While the Green's function used here is somewhat different from Wang et al., J. Phys. D 29, 1649 (1996), the resulting analytic expression for the potential field is in exact agreement with their result. Selected results for equispaced electrodes with equal widths are compared with Wang et al., J. Phys. D 29, 1649 (1996). The analytic solution is employed to study the effects of electrode spacing and electrode width on the gradient in electric field intensity. Results show that the magnitude in the gradient in the electric field intensity exhibited the expected dependence on the applied voltage; however, the dependence on electrode width was found to be on the order of the electrode width squared. Results to explore the effects of electrode spacing show that as the spacing is reduced below two electrode widths the magnitude of the gradient increases exponentially. PMID- 11497729 TI - Transmission delay times of localized waves. AB - We investigate the effects of wave localization on the delay time tau (frequency sensitivity of the scattering phase shift) of a wave transmitted through a disordered waveguide. Localization results in a separation tau=chi+chi(') of the delay time into two independent but equivalent contributions, associated to the left and right end of the waveguide. For N=1 propagating modes, chi and chi(') are identical to half the reflection delay time of each end of the waveguide. In this case the distribution function P(tau) in an ensemble of random disorder can be obtained analytically. For N>1 propagating modes the distribution function can be approximated by a simple heuristic modification of the single-channel problem. We find a strong correlation between channels with long reflection delay times and the dominant transmission channel. PMID- 11497730 TI - Interactions of vector solitons. AB - In this paper, we study the interaction of two widely separated vector solitons in the nonintegrable coupled nonlinear Schrodinger (NLS) equations. Using a modification of Karpman-Solov'ev perturbation method, we derive dynamical equations for the evolution of both solitons' internal parameters. We show that these dynamical equations allow fixed points that correspond to stationary two vector-soliton bound states if these solitons have the same phase in one component (same sign) and pi-phase difference in the other component (opposite sign). However, linear stability analysis indicates that these bound states are always unstable due to a phase-related unstable eigenvalue. We also investigate vector-soliton interactions and show that, in contrast to soliton interactions in the single NLS equation, vector solitons repel or attract each other depending not only on their relative phases but also on their initial position separation. Lastly, interaction of an arbitrary number of vector solitons is also studied in brief. All our analytical results are supported by direct numerical simulations. PMID- 11497731 TI - Compacton solutions in a class of generalized fifth-order Korteweg-de Vries equations. AB - Solitons play a fundamental role in the evolution of general initial data for quasilinear dispersive partial differential equations, such as the Korteweg-de Vries (KdV), nonlinear Schrodinger, and the Kadomtsev-Petviashvili equations. These integrable equations have linear dispersion and the solitons have infinite support. We have derived and investigate a new KdV-like Hamiltonian partial differential equation from a four-parameter Lagrangian where the nonlinear dispersion gives rise to solitons with compact support (compactons). The new equation does not seem to be integrable and only mass, momentum, and energy seem to be conserved; yet, the solitons display almost the same modal decompositions and structural stability observed in integrable partial differential equations. The compactons formed from arbitrary initial data, are nonlinearly self stabilizing, and maintain their coherence after multiple collisions. The robustness of these compactons and the inapplicability of the inverse scattering tools, that worked so well for the KdV equation, make it clear that there is a fundamental mechanism underlying the processes beyond integrability. We have found explicit formulas for multiple classes of compact traveling wave solutions. When there are more than one compacton solution for a particular set of parameters, the wider compacton is the minimum of a reduced Hamiltonian and is the only one that is stable. PMID- 11497732 TI - Universal photonic tunneling time. AB - We consider photonic tunneling through evanescent regions and obtain general analytic expressions for the transit (phase) time tau (in the opaque barrier limit) in order to study the recently proposed "universality" property according to which tau is given by the reciprocal of the photon frequency. We consider different physical phenomena (corresponding to performed experiments) and show that such a property is only an approximation. In particular, we find that the "correction" factor is a constant term for total internal reflection and quarter wave photonic band gap, while it is frequency dependent in the case of an undersized waveguide and distributed Bragg reflector. The comparison of our predictions with the experimental results shows quite good agreement with observations and reveals the range of applicability of the approximate "universality" property. PMID- 11497733 TI - Investigation of transition radiation from a regular-roughness interface. AB - Transition radiation at the interface between two media having regular inhomogeneities is considered in the framework of perturbation theory, assuming that the dielectric constants of the two media differ insignificantly from each other. The advantage of this approach is that no limitations exist on the characteristic size of the inhomogeneities. The general case of a surface with two-dimensional roughness is considered. Attention is paid to radiation characteristics that are general for periodic structures, independently of their particular features. The physical picture of the radiation from a rough interface is determined by both longitudinal and transverse effects. The case of normal incidence of an electron on a planar interface having either single or multiple rectangular lugs (diffraction grating) is analyzed in detail. For the single lug infinite in one dimension, simple expressions for transition radiation intensity are obtained, for both relativistic and nonrelativistic electrons. They demonstrate that corrections to the plane-surface intensity become significant at small radiation angles. In this case the radiation is also completely unpolarized, i.e., the spectral energy density of the perpendicular component has the same order of magnitude as that of the parallel component. Consideration of these issues is important since rough surfaces, as compared to planar surfaces, give a number of additional results that may be used for the analysis of surface irregularities. PMID- 11497734 TI - Multipulses in discrete Hamiltonian nonlinear systems. AB - In this work, the behavior of multipulses in discrete Hamiltonian nonlinear systems is investigated. The discrete nonlinear Schrodinger equation is used as the benchmark system for this study. A singular perturbation methodology as well as a variational approach are implemented in order to identify the dominant factors in the discrete problem. The results of the two methodologies are shown to coincide in assessing the interplay of discreteness and exponential tail-tail pulse interaction. They also allow one to understand why, contrary to what is believed for their continuum siblings, discrete systems can sustain (static) multipulse configurations, a conclusion that is subsequently verified by numerical experiment. PMID- 11497735 TI - Influence of the relative refractive index on the depolarization of multiply scattered waves. AB - Using the theory of radiative transfer, we investigate the interaction between polarized waves and a multiple scattering medium as functions of the relative index of refraction. To study this problem, we consider circularly and linearly polarized continuous waves incident upon a medium containing spherical scatterers. With an accurate spectral method, we compute the transmitted Stokes parameters through media containing different sized scatterers and different indices of refraction. Our numerical results show that the circular depolarization length exhibits a strong dependence on the relative index of refraction, while the linear depolarization length does not. PMID- 11497736 TI - Interaction between sine-Gordon breathers. AB - Using the exact breather lattice solution of the sine-Gordon equation, we obtain the asymptotic interaction between two breathers. We identify the exponential dependence of the interaction on the breather separation as well as its power-law dependence on the breather frequency. Numerical simulation of the breather lattice demonstrates its instability. However, stabilization of such structures is found to be feasible through ac driving and damping. Finally, other limits of the original periodic solution are traced, obtaining the leading-order terms in the interaction between "pseudosphere" solutions and kink-antikink pairs. PMID- 11497737 TI - Dynamics of solitons and quasisolitons of the cubic third-order nonlinear Schrodinger equation. AB - The dynamics of soliton and quasisoliton solutions of the cubic third-order nonlinear Schrodinger equation is studied. Regular solitons exist due to a balance between the nonlinear terms and (linear) third-order dispersion; they are not important at small alpha(3) (alpha(3) is the coefficient in the third derivative term) and vanish at alpha(3)-->0. The most essential, at small alpha(3), is a quasisoliton emitting resonant radiation (resonantly radiating soliton). Its relationship with the other (steady) quasisoliton, called embedded soliton, is studied analytically and also in numerical experiments. It is demonstrated that the resonantly radiating solitons emerge in the course of nonlinear evolution, which shows their physical significance. PMID- 11497738 TI - Rigorous classical-mechanical derivation of a multiple-copy algorithm for sampling statistical mechanical ensembles. AB - We derive a rigorous, multiple-copy simulation algorithm that is formally equivalent to conventional classical molecular dynamics for an ensemble of systems, but may be used for rapid geometry optimizations. The derivation is accomplished by starting from an ensemble of copies of the entire system and applying a point coordinate transformation to a large subsystem defined as the bath. After the transformation, each atom of the bath is described by one "major" set of coordinates located at the average position of the ensemble of equivalent atoms and a set of "minor" coordinates that when combined with the "major" coordinates represent exact dynamics. Neglecting the "minor" set of coordinates results in a Hamiltonian and a probability density equivalent to those used in existing multiple-copy methods. Neglecting Hamilton's equations of motion for the minor variables gives the equations of motion for locally enhanced sampling. Numerical tests indicate that the algorithm can recover exact molecular dynamics of the ensemble, conventional multiple-copy dynamics, or results of intermediate accuracy. Thus, the algorithm provides a rigorous basis for multiple-copy dynamics, resolves many of the uncertainties associated with their current implementations, and offers the potential for calculating ensemble average properties in conjunction with finding a system's global minimum energy geometry. PMID- 11497739 TI - Parity problem with a cellular automaton solution. AB - The parity of a bit string of length N is a global quantity that can be efficiently computed using a global counter in O(N) time. But is it possible to find the parity using cellular automata with a set of local rule tables without using any global counter? Here, we report a way to solve this problem using a number of r=1 binary, uniform, parallel, and deterministic cellular automata applied in succession for a total of O(N2) time. PMID- 11497740 TI - Atomic charges in molecular mechanical force fields: a theoretical insight. AB - Based on quantum theory of the Coulombic interactions between a molecule and its surrounding molecules, a theoretical derivation is presented to obtain an atomic charge model. The charge model shows that the appropriate atomic charge, used for example in molecular mechanical force fields, is simply the average value of the electrostatic potential (ESP) derived charge for an isolated molecule and that obtained for the molecule in the interaction system. Computational scheme to calculate the ESP derived atomic charges of a molecule in the interaction system is presented. The method is applied to two common liquids with associated hydrogen bonds: water and methanol. The obtained theoretical atomic charges are similar to those found in the common interaction potential models, such as: SPC, TIP3P, OPLS, etc. However, for methanol they differ considerably from those obtained using the restrained ESP method. PMID- 11497741 TI - Spectra of "real-world" graphs: beyond the semicircle law. AB - Many natural and social systems develop complex networks that are usually modeled as random graphs. The eigenvalue spectrum of these graphs provides information about their structural properties. While the semicircle law is known to describe the spectral densities of uncorrelated random graphs, much less is known about the spectra of real-world graphs, describing such complex systems as the Internet, metabolic pathways, networks of power stations, scientific collaborations, or movie actors, which are inherently correlated and usually very sparse. An important limitation in addressing the spectra of these systems is that the numerical determination of the spectra for systems with more than a few thousand nodes is prohibitively time and memory consuming. Making use of recent advances in algorithms for spectral characterization, here we develop methods to determine the eigenvalues of networks comparable in size to real systems, obtaining several surprising results on the spectra of adjacency matrices corresponding to models of real-world graphs. We find that when the number of links grows as the number of nodes, the spectral density of uncorrelated random matrices does not converge to the semicircle law. Furthermore, the spectra of real-world graphs have specific features, depending on the details of the corresponding models. In particular, scale-free graphs develop a trianglelike spectral density with a power-law tail, while small-world graphs have a complex spectral density consisting of several sharp peaks. These and further results indicate that the spectra of correlated graphs represent a practical tool for graph classification and can provide useful insight into the relevant structural properties of real networks. PMID- 11497742 TI - Damage-spreading phase and damage-frozen phase in a solid-on-solid model. AB - Based on the recently suggested scaling ansatz [Phys. Rev. E 62, 3376 (2000)] for damage spreading in the surface roughening phenomenon, the characteristics of the damage-spreading phase and damage-frozen phase in a two-dimensional solid-on solid model that has a roughening transition at T=T(R) are studied. In the damage spreading phase, which exists for T>T(R), the average vertical damage-spreading distance d(perpendicular)(d(//)=0,L,T) and the average lateral damage-spreading distance D(//)(L,T) are shown to satisfy d(perpendicular)(d(//)=0,L,T) approximately ln L and D(//)(L,T) approximately L, respectively. In the damage frozen phase, which exists for Tinfinity,T) approximately finite and D(//)(L- >infinity,T) approximately finite. From these results it is concluded that the damage-spreading phase describes the surface roughening phase well and the damage frozen phase describe the smooth phase well. PMID- 11497743 TI - Propagation failure in discrete bistable reaction-diffusion systems: theory and experiments. AB - Wave front propagation failure is investigated in discrete bistable reaction diffusion systems. We present a theoretical approach including dissipative effects and leading to an analytical expression of the critical coupling beyond which front propagation can occur as a function of the nonlinearity threshold parameter. Our theoretical predictions are confirmed by numerical simulations and experimental results on an equivalent electrical diffusive lattice. PMID- 11497744 TI - Area-slope relation in a simple erosion model. AB - We discuss area-slope relation of a landform evolved by a simple lattice model of an erosion process. Observing a steady state of the model, where river networks on the surface are stationary, we present the relation between the power exponent of the area-slope relation and the parameters of the erosion process of the model. We show how to determine the parameters of the erosion process from observations of landform by making use of the area-slope relation. This method may be useful for morphological simulations of the landforms. PMID- 11497745 TI - Probabilistic properties of wavelets in kinetic surface roughening. AB - Using the data of a recent numerical simulation [M. Ahr and M. Biehl, Phys. Rev. E 62, 1773 (2000)] of homoepitaxial growth it is shown that the observed probability distribution of a wavelet based measure of the growing surface roughness is consistent with a stretched log-normal distribution and the corresponding branching dimension depends on the level of particle desorption. PMID- 11497746 TI - Dilute Bose gas in two dimensions: density expansions and the Gross-Pitaevskii equation. AB - A dilute homogeneous two-dimensional (2D) Bose gas at zero temperature is studied with the method developed earlier by the authors. This method allows for considering renormalization of an arbitrary pairwise potential in a self consistent manner, without the pseudopotential delta-function representation. Low density expansions are derived for the chemical potential, ground-state energy, pair distribution function, kinetic and interaction energies. The expansion parameter is found to be a dimensionless in-medium scattering amplitude u obeying the equation 1/u+ln u=-ln(na(2)pi)-2gamma, where na(2) and gamma are the gas parameter and the Euler constant, respectively. It is shown that the ground-state energy is mostly kinetic in the low-density limit. This result does not depend on a specific form of the pairwise interaction potential, contrary to the 3D case. A new form of the 2D Gross-Pitaevskii equation is proposed within our scheme. PMID- 11497747 TI - Level spacings and periodic orbits. AB - Starting from a semiclassical quantization condition based on the trace formula, we derive a periodic-orbit formula for the distribution of spacings of eigenvalues with k intermediate levels. Numerical tests verify the validity of this representation for the nearest-neighbor level spacing (k=0). In a second part, we present an asymptotic evaluation for large spacings, where consistency with random matrix theory is achieved for large k. We also discuss the relation with the method of Bogomolny and Keating [Phys. Rev. Lett. 77, 1472 (1996)] for two-point correlations. PMID- 11497748 TI - Vortex pairs in viscoelastic Couette-Taylor flow. AB - In experiments on dilute polymers between rotating cylinders Groisman and Steinberg [Phys. Rev. Lett. 78, 1460 (1997)] observed the formation of vortices that were not equidistantly spaced but rather paired up in what they called "diwhirls." We calculate these states within an Oldroyd-B model with parameters adapted to the experiment and find good agreement with the observed characteristics of the diwhirls. PMID- 11497749 TI - Phase transitions and critical phenomena in the liquid bridge under lateral acceleration. AB - We study the shape of a liquid body under the influence of a lateral body force in a thin liquid bridge. The shape of the liquid body changes continuously or discontinuously depending on the volume of liquid body. We show that the transition from discontinuous to continuous change is described by the volume induced phase transition driven by the lateral body force. The order parameter of the phase transition is the amount of shift of the center of mass of liquid body. Critical exponents of a mean-field model have been obtained by numerical calculation. We provide the validity of this critical behavior through the m(4) model of Landau theory of phase transition. PMID- 11497750 TI - Power-law scaling in Benard-Marangoni convection at large Prandtl numbers. AB - Benard-Marangoni convection at large Prandtl numbers is found to exhibit steady (nonturbulent) behavior in numerical experiments over a very wide range of Marangoni numbers Ma far away from the primary instability threshold. A phenomenological theory, taking into account the different character of thermal boundary layers at the bottom and at the free surface, is developed. It predicts a power-law scaling for the nondimensional velocity (Peclet number) and heat flux (Nusselt number) of the form Pe approximately Ma(2/3), Nu approximately Ma(2/9). This prediction is in good agreement with two-dimensional direct numerical simulations up to Ma=3.2 x 10(5). PMID- 11497751 TI - Waves in strongly magnetized relativistic plasmas: generally covariant approach. AB - A dispersion relation for long waves in strongly magnetized multifluid plasma in a curved spacetime is derived in a covariant form. A generally covariant form for the ray equations is obtained. The results are applicable to ray propagation in relativistic plasmas in the vicinity of strongly gravitating (black holes) or rapidly rotating (pulsars) systems. PMID- 11497752 TI - Subsystem purity as an enforcer of entanglement. AB - We show that entanglement can always arise in the interaction of an arbitrarily large system in any mixed state with a single qubit in a pure state. This small initial purity is enough to enforce entanglement even when the total entropy is close to maximum. We demonstrate this feature using the Jaynes-Cummings interaction of a two-level atom in a pure state with a field in a thermal state at an arbitrarily high temperature. We find the time and temperature variation of a lower bound on the amount of entanglement produced and study the classical correlations quantified by the mutual information. PMID- 11497753 TI - Quantum state reconstruction of the single-photon Fock state. AB - We have reconstructed the quantum state of optical pulses containing single photons using the method of phase-randomized pulsed optical homodyne tomography. The single-photon Fock state 1> was prepared using conditional measurements on photon pairs born in the process of parametric down-conversion. A probability distribution of the phase-averaged electric field amplitudes with a strongly non Gaussian shape is obtained with the total detection efficiency of (55+/-1)%. The angle-averaged Wigner function reconstructed from this distribution shows a strong dip reaching classically impossible negative values around the origin of the phase space. PMID- 11497754 TI - Measurement of one-particle correlations and momentum distributions for trapped 1D gases. AB - Van Hove's theory of scattering of probe particles by a macroscopic target is generalized so as to relate the differential cross section for atomic ejection via stimulated Raman transitions to one-particle momentum-time correlations and momentum distributions of 1D trapped gases. This method is well suited to probing the longitudinal momentum distributions of 1D gases in situ, and examples are given for bosonic and fermionic atoms. PMID- 11497755 TI - Phase-fluctuating 3D Bose-Einstein condensates in elongated traps. AB - We find that in very elongated 3D trapped Bose gases, even at temperatures far below the BEC transition temperature T(c), the equilibrium state will be a 3D condensate with fluctuating phase (quasicondensate). At sufficiently low temperatures the phase fluctuations are suppressed and the quasicondensate turns into a true condensate. The presence of the phase fluctuations allows for extending thermometry of Bose-condensed gases well below those established in current experiments. PMID- 11497756 TI - Angular momentum transport in thin accretion disks and intermittent accretion. AB - The plasma modes, transporting angular momentum in accretion disks, under minimally restrictive conditions when the magnetic energy density is significant relative to the thermal energy density, are shown to be singular if the ideal MHD approximation is adopted. A similarity with the modes producing magnetic reconnection in current carrying plasmas is established. The combined effects of finite plasma temperature, of plasma compressibility, of the gradient of the rotation frequency, and of appropriate transport processes (outside ideal MHD) are involved in the onset of these nonaxisymmetric and locally corotating modes. PMID- 11497757 TI - New supernova limit on large extra dimensions: bounds on Kaluza-Klein graviton production. AB - If large extra dimensions exist in nature, supernova (SN) cores will emit large fluxes of Kaluza-Klein gravitons, producing a cosmic background of these particles with energies and masses up to about 100 MeV. Radiative decays then give rise to a diffuse cosmic gamma-ray background with E(gamma) approximately less than 100 MeV which is well in excess of the observations if more than 0.5% 1% of the SN energy is emitted into the new channel. For two extra dimensions we derive a conservative bound on their radius of R approximately less than 0.9 x 10(-4) mm; for three extra dimensions it is R approximately less than 1.9 x 10( 7) mm. PMID- 11497758 TI - Nonthermal production of gravitinos and inflatinos. AB - We explicitly calculate nonthermal gravitino production during the preheating period in the inflationary Universe. Contrary to earlier investigations, we consider a two-field model to separate the mechanisms of supersymmetry breaking and inflation. We show that the superpartner of the inflaton is significantly generated, while the gravitino production is considerably smaller. Nonthermal production of gravitinos seems thus less worrisome than recently claimed. PMID- 11497759 TI - Relativistic corrections to the electric polarizability of compound systems. AB - The widely used relation alpha; = alpha(0)+Deltaalpha for the electric polarizability, where Deltaalpha is proportional to the r.m.s. charge radius of the system, is revisited within a simple model of two spinless particles bound electromagnetically. A complete set of the relativistic corrections to alpha(0) following from the Breit Hamiltonian is obtained. It is shown that these corrections modify the result for alpha; in comparison to that obtained with alpha(0) taken in the nonrelativistic approximation. We propose that the same situation can take place in the more complicated case of hadrons. Special attention is devoted to the correct definition of the center-of-mass coordinates which are found to be very important. PMID- 11497760 TI - Direct observation of a rho decay of the D13(1520) baryon resonance. AB - The reaction gammap-->pi(+)pi(0)n has been measured at MAMI for photon energies up to 820 MeV. Invariant mass spectra of the particles in the final state (pi(+)n), (pi(0)n), (pi(+)pi(0)) have been determined for several bins of incident photon energy. Differences in pi(+)pi(0) and simultaneously measured pi(0)pi(0) invariant mass distributions are assigned to a rho branch of the D13(1520) nucleon resonance. PMID- 11497761 TI - Transport theory for a two-flavor color superconductor. AB - QCD with two light-quark flavors at high baryonic density is a color superconductor. The diquark condensate breaks the SU(3) gauge symmetry down to an SU(2) subgroup. We study thermal fluctuations of the superconductor for temperatures below the gap. These are described by a simple transport equation. In the collisionless limit and close to equilibrium, it gives rise to the "hard superconducting loop" effective theory for the SU(2) gauge fields. This theory describes Debye screening and Landau damping of the gauge fields in the presence of the diquark condensate. We explain how our effective theory follows to one loop order from quantum field theory. Our approach provides a convenient starting point for the computation of transport coefficients of the two-flavor color superconductor. PMID- 11497763 TI - Ambiguities in the implementation of the impulse approximation for the response of many-fermion systems. AB - Within the impulse approximation, the response of a many-body system at large momentum transfer can be directly related to ground state properties. Although the physics assumptions underlying impulse approximation are well defined, their implementation involves ambiguities that may cause significant differences in the calculated responses. We show that, while minimal use of the impulse approximation assumptions naturally leads to write the response in terms of the spectral function, the alternative definition in terms of the momentum distribution involves a more extended use of the same assumptions. PMID- 11497764 TI - Analytic description of critical point nuclei in a spherical-axially deformed shape phase transition. AB - An approximate solution at the critical point of the spherical to axially deformed shape phase transition in nuclei is presented. The eigenvalues of the Hamiltonian are expressed in terms of zeroes of Bessel functions of irrational order. PMID- 11497765 TI - Empirical realization of a critical point description in atomic nuclei. AB - It is shown that (152)Sm and other N = 90 isotones are the first empirical manifestation of the newly predicted analytic description of nuclei at the critical point of a vibrator to axial rotor phase transition. PMID- 11497766 TI - Microscopic structure of fundamental excitations in N = Z nuclei. AB - An extended mean-field model is presented that describes states of different isospin in odd-odd and even-even nuclei. Excitation energies of the T = 1 states in even-even as well as T = 0 and T = 1 states in odd-odd N = Z nuclei are calculated. It is shown that the structure of these states can be determined in a consistent manner when both isoscalar and isovector pairing collectivity as well as isospin projection (treated here within the isocranking approximation) are taken into account. In particular, in odd-odd N = Z nuclei, the interplay between quasiparticle excitations (relevant for the case of T = 0 states) and isorotations (relevant for the case of T = 1 states) explains the near degeneracy of these states. PMID- 11497767 TI - Gamma-ray multiplicities and fission modes in (208)Pb((18)O,f). AB - Two components in the M(gamma)(M) distribution were established in detailed measurements of mean gamma-ray multiplicities from fission fragments of (226)Th. For the first time in the M(gamma)(M) dependencies we were able to distinguish two components associated with primary and the final (after the neutron evaporation) fission fragments, and show that at the scission point M(gamma) is extremely sensitive to symmetric and asymmetric modes of fission. Theoretical calculations of the pre-scission shapes of the fissioning nuclei confirm our conclusions. PMID- 11497768 TI - Dynamics and post-collision interaction effects in two electron decay from the Xenon 4d hole. AB - Two Auger electrons, one very slow, one fast, have been detected in coincidence following near threshold 4d photoionization of the Xe atom. The distribution in the energy the two electrons share has been measured for the first time revealing the presence of post-collision interaction effects that provide unique information on the decay dynamics of the 4d hole. Analysis of the distorted line shapes indicates that the dominant process is decay of Xe+(4d(-1)) to Xe3+ through cascade emission of a zero kinetic energy Auger electron followed by a fast Auger electron. The widths of the intermediate Xe2+* states are estimated to be about 60 meV. PMID- 11497769 TI - Photoelectron spectra from multiple ionization of atoms in ultra-intense laser pulses. AB - We address the question of the energy and angular distributions of the photoelectrons ejected from rare gas atoms submitted to ultra-intense infrared laser pulses, with peak intensities I(max) approximately 10(18) W/cm (2). Several unsolved issues regarding the angular distributions of the photoelectrons are analyzed. We believe that our results should help to trigger new investigations. PMID- 11497770 TI - Photoabsorption spectra of Na(+)(n) clusters: thermal line-broadening mechanisms. AB - Photoabsorption cross sections of small sodium cluster cations ( Na(+)(n), n = 3, 5, 7, and 9) were calculated at various temperatures with the time-dependent local-density approximation in conjunction with ab initio molecular dynamics simulations, yielding spectra that agree with measured ones without ad hoc line broadening or renormalization. Three thermal line-broadening mechanisms are revealed: (I) lifting of level degeneracies caused by symmetry-breaking ionic motions, (II) oscillatory shifts of the entire spectrum caused by breathing vibrations, and (III) cluster structural isomerizations. PMID- 11497762 TI - Measurement of the midrapidity transverse energy distribution from square root of [(s)NN] = 130 GeV Au + Au collisions at RHIC. AB - The first measurement of energy produced transverse to the beam direction at the Relativistic Heavy-Ion Collider at Brookhaven National Laboratory is presented. The midrapidity transverse energy density per participating nucleon rises steadily with the number of participants, closely paralleling the rise in charged particle density, such that / remains relatively constant as a function of centrality. The energy density calculated via Bjorken's prescription for the 2% most central Au+Au collisions at square root[s(NN)] = 130 GeV is at least epsilon(Bj) = 4.6 GeV/fm(3), which is a factor of 1.6 larger than found at sqrt[s(NN)] = 17.2 GeV ( Pb+Pb at CERN). PMID- 11497771 TI - Interferometric pump-probe study of intense field excitation of sapphire. AB - Under intense optical excitation, the polarizability of transparent optical materials has a significant third order, nonlinear contribution. This manifests itself in a wide variety of effects, which, under certain conditions, are collectively referred to as white light continuum generation. We report on a new approach that isolates, for the first time, some of these effects. PMID- 11497772 TI - Effect of noise on the relaxation to an invariant probability measure of nonhyperbolic chaotic attractors. AB - We study the influence of external noise on the relaxation to an invariant probability measure for two types of nonhyperbolic chaotic attractors, a spiral (or coherent) and a noncoherent one. We find that for the coherent attractor the rate of mixing changes under the influence of noise, although the largest Lyapunov exponent remains almost unchanged. A mechanism of the noise influence on mixing is presented which is associated with the dynamics of the instantaneous phase of chaotic trajectories. This also explains why the noncoherent regime is robust against the presence of external noise. PMID- 11497773 TI - Coherent structures and entropy in constrained, modulationally unstable, nonintegrable systems. AB - Many studies have shown that nonintegrable systems with modulational instabilities constrained by more than one conservation law exhibit universal long time behavior involving large coherent structures in a sea of small fluctuations. We show how this behavior can be explained in detail by simple thermodynamic arguments. PMID- 11497774 TI - Coherent vortex extraction in 3D turbulent flows using orthogonal wavelets. AB - This Letter presents a wavelet technique for extracting coherent vortices from three-dimensional turbulent flows, which is applied to a homogeneous isotropic turbulent flow at resolution N = 256(3). The coherent flow is reconstructed from only 3%N wavelet coefficients that retain the vortex tubes, and 98.9% of the energy with the same k(-5/3) spectrum as the total flow. In contrast, the remaining 97%N wavelet coefficients correspond to the incoherent flow which is structureless, decorrelated, and whose effect can therefore be modeled statistically. PMID- 11497775 TI - Fingering of chemical fronts in porous media. AB - The influence of chemical reactions on the hydrodynamical fingering instability is analyzed for miscible systems in porous media. Using a realistic reaction scheme, it is shown that the stability of chemical fronts towards density fingering crucially depends on the width and the speed of the front which are functions of chemical parameters. The major difference between the pure and chemically driven fingering is that, in the presence of chemical reactions, the dispersion curves do not vary in time which has important practical experimental consequences. Good agreement with recent experimental data is found. PMID- 11497776 TI - Giant deformations of a liquid-liquid interface induced by the optical radiation pressure. AB - Because of the small momentum of photons, very intense fields are generally required to bend a liquid interface with the optical radiation pressure. We explore this issue in a near-critical phase-separated liquid mixture to vary continuously the meniscus softness by tuning the temperature. Low power continuous laser waves become sufficient to induce huge stationary bulges. Using the beam size to build an "optical" Bond number, Bo, we investigate the crossover from low to large Bo. The whole set of data collapses onto a single master curve which illustrates the universality of the phenomenon. PMID- 11497777 TI - Shear-dependent boundary slip in an aqueous Newtonian liquid. AB - We report direct measurements of hydrodynamic drainage forces, which show clear evidence of boundary slip in a Newtonian liquid. The degree of boundary slip is found to be a function of the liquid viscosity and the shear rate, as characterized by the slip length, and is up to approximately 20 nm. This has implications for confined biological systems, the permeability of microporous media, and for the lubrication of nanomachines, and will be important in the microcontrol of liquid flow. We also show that current theories of slip do not adequately describe the experimental data. PMID- 11497778 TI - First measurements of the unique influence of spin on the energy loss of ultrarelativistic electrons in strong electromagnetic fields. AB - Although some authors have claimed that the effect is not detectable, we show experimentally for the first time that as the quantum parameter chi grows beyond 1, an increasingly large part of the hard radiation emitted arises from the spin of the electron. Results for the energy loss of electrons in the energy range 35 243 GeV incident on a W single crystal are presented. Close to the axial direction the strong electromagnetic fields induce a radiative energy loss which is significantly enhanced compared to incidence on an amorphous target. In such continuously strong fields, the radiation process is highly nonperturbative for ultrarelativistic particles and a full quantum description is needed. The remarkable effect of spin flips and the energy loss is connected to the presence of a field comparable in magnitude to the Schwinger critical field, E0 = m(2)c(3)/ePlanck's over 2pi, in the rest frame of the emitting electron. PMID- 11497779 TI - Ion motion synchronization in an ion-trap resonator. AB - Using a new type of ion trap, we demonstrate that the length of a packet of charged particles oscillating between two electrostatic mirrors will remain constant under special conditions. The effect can be understood in terms of phase synchronization, where, in a rather counterintuitive way, the repulsive Coulomb interaction between the ions actually holds the packet together. Application of this effect for mass spectrometry is discussed. PMID- 11497780 TI - Neoclassical radial electric field and transport with finite orbits. AB - Neoclassical transport in a toroidal plasma with finite ion orbits is studied, including for the first time the self-consistent radial electric field. Using a low-noise deltaf particle simulation, we demonstrate that a deep electric-field well develops in a region with a steep density gradient, because of the self collision-driven ion flux. We find that the electric field agrees with the standard neoclassical expression, when the toroidal rotation is zero, even for a steep density gradient. Ion thermal transport is modified by the electric-field well in a way which is consistent with the orbit squeezing effect, but smoothed by the finite orbits. PMID- 11497781 TI - Pressure dependent incommensuration in Rb-IV. AB - Rb-IV is found to have an incommensurate composite structure, comprising a tetragonal host framework and a simple body-centered tetragonal guest. This does not have the unexpectedly short Rb-Rb distances of the previously reported structure [U. Schwarz et al., Phys. Rev. Lett. 83, 4085 (1999)]. The ratio of the c-axis lattice parameters is strongly pressure dependent and approaches the commensurate value of 5/3 at the transition to phase V. A reversible broadening of the guest structure is observed below 16.5(2) GPa. PMID- 11497782 TI - Topological description of the aging dynamics in simple glasses. AB - We numerically investigate the aging dynamics of a monatomic Lennard-Jones glass, focusing on the topology of the potential energy landscape which, to this aim, has been partitioned in basins of attraction of stationary points (saddles and minima). The analysis of the stationary points visited during the aging dynamics shows the existence of two distinct regimes: (i) at short times the system visits basins of saddles whose energies and orders decrease with t; (ii) at long times the system mainly lies in basins pertaining to minima of slowly decreasing energy. The long time dynamics can be represented by a simple random walk on a network of minima with a jump probability proportional to the inverse of the waiting time. PMID- 11497783 TI - Capillary condensation in disordered porous materials: hysteresis versus equilibrium behavior. AB - We study the interplay between hysteresis and equilibrium behavior in capillary condensation of fluids in mesoporous disordered materials via a mean-field density functional theory of a disordered lattice-gas model. The approach reproduces all major features observed experimentally. We show that the simple van der Waals picture of metastability fails due to the appearance of a complex free-energy landscape with a large number of metastable states. In particular, hysteresis can occur both with and without an underlying equilibrium transition, and thermodynamic consistency is not satisfied along the hysteresis loop. PMID- 11497784 TI - Two-dimensional nucleation of ice from supercooled water. AB - We report the temperature dependent nucleation rates of ice from single water drops supporting aliphatic alcohol Langmuir films. Analysis in the context of a classical theory of heterogeneous nucleation suggests that the critical nucleus is essentially a monolayer, and that the rate-limiting steps in these nucleation processes are therefore not merely influenced by, but instead dictated by, the physics of the water-alcohol interface. Consequently, reduced dimensionality may be much more important in heterogeneous nucleation than has previously been believed. PMID- 11497785 TI - Melting mechanisms at the limit of superheating. AB - The atomic-scale details during melting of a surface-free Lennard-Jones crystal were monitored using molecular dynamics simulations. Melting occurs when the superheated crystal spontaneously generates a sufficiently large number of spatially correlated destabilized particles that simultaneously satisfy the Lindemann and Born instability criteria. The accumulation and coalescence of these internal local lattice instabilities constitute the primary mechanism for homogeneous melt nucleation inside the crystal, in lieu of surface nucleation for equilibrium melting. The vibrational and elastic lattice instability criteria as well as the homogeneous nucleation theory all coincide in determining the superheating limit. PMID- 11497786 TI - Three-dimensional elastic compatibility and varieties of twins in martensites. AB - We model a cubic-to-tetragonal martensitic transition by a Ginzburg-Landau free energy in the symmetric strain tensor. We show in three dimensions (3D) that solving the St. Venant compatibility relations for strain, treated as independent field equations, generates three anisotropic long-range potentials between the two order parameter components. These potentials encode 3D discrete symmetries, express the energetics of lattice integrity, and determine 3D textures. Simulation predictions include twins with temperature-varying orientation, helical twins, competing metastable states, and compatibility-induced elastic frustration. Our approach also applies to improper ferroelastics. PMID- 11497787 TI - Role of tilt order in the asymmetric ripple phase of phospholipid bilayers. AB - We present the electron density map of the asymmetric ripple phase of dilauroylphosphatidylcholine. We find that the primary feature characterizing the "asymmetry" of the rippled bilayers is the difference in the bilayer thickness in the two arms, and not the asymmetry of the bilayer height profile as is generally assumed. This difference in the bilayer thickness can be attributed to a mean tilt of the hydrocarbon chains of the lipid molecules along the direction of the ripple wave vector. We propose a Landau theory for this phase which takes into account the anisotropic elastic properties of a bilayer with tilt order. PMID- 11497788 TI - Molecular origin of thermal diffusion in benzene + cyclohexane mixtures. AB - The isotope effect in thermal diffusion (Soret effect) of benzene+cyclohexane mixtures has been investigated by a holographic grating technique. The Soret coefficient can be split into additive contributions. One contribution, the isotope effect, stems from the differences of both mass and moment of inertia, and is independent of composition. An additional "chemical" contribution depends on concentration and even changes its sign at a benzene mole fraction x(benz) approximately 0.7. The mass effect is in agreement with molecular dynamics calculations: the heavier component migrates to the cold side. PMID- 11497789 TI - Surface-reconstruction-switched adsorbate photofragmentation dynamics. AB - Energy-resolved angular distributions of neutral fragments ejected during photoinduced electron transfer reaction of CH3Br on GaAs(100) exhibit three distinct methyl-radical ejection channels. These undergo marked changes when the termination is switched from the Ga-rich c(8 x 2) to the As-rich c(2 x 8). Our observations are consistent with a strong adsorption-site dependence of the dynamics. PMID- 11497790 TI - Spatiotemporal distribution of nucleation events during crystal growth. AB - We consider irreversible second-layer nucleation that occurs when two adatoms on a terrace meet. We solve the problem analytically in one dimension for zero and infinite step-edge barriers, and numerically for any value of the barriers in one and two dimensions. For large barriers, the spatial distribution of nucleation events strongly differs from rho(2), where rho is the stationary adatom density in the presence of a constant flux. Theories of the nucleation rate omega based on the assumption that it is proportional to rho(2) are shown to overestimate omega by a factor proportional to the number of times an adatom diffusing on the terrace visits an already visited lattice site. PMID- 11497791 TI - Oxygen dissociation at Pt steps. AB - Using scanning tunneling microscopy, thermal energy atom scattering, and density functional theory we have characterized O (2) dissociation on Pt(111) stepped surfaces at the atomic scale. The most reactive site is at the top of the Pt steps. In both the molecular precursor state (MPS) and the transition state (TS), the O (2) has its axis aligned parallel to the step edge. Controlled step decoration with Ag monatomic chains was used to locally tune the reactivity of Pt step sites. The enhanced reactivity at the Pt step sites is not caused by a decrease of the local dissociation barriers from the MPS but is related to a stabilization of both the MPS and TS. PMID- 11497792 TI - Structure of the Ba-induced Si(111)- (3 x 2) reconstruction. AB - The Ba/Si(111) surface, previously known as a 3 x 1 phase, is found to have a 3 x 2 periodicity and a semiconducting band gap. The substrate reconstructs into the honeycomb chain-channel (HCC) structure with Ba atoms in the channel, as in the alkali-metal-induced Si(111)-(3 x 1). However, the metal coverage is determined to be 1/6 monolayers, half the alkali-metal coverage. We propose that the structure and the metal coverage determined for the Ba adsorbate is universal for other alkaline-earth-metal adsorbates. With the alkali-metal-induced 3 x 1 case, our results lead to a rule that one donated electron per 3 x 1 surface unit is necessary to stabilize the HCC reconstruction of Si. PMID- 11497793 TI - Anomalous isotopic effect near the charge-ordering quantum criticality. AB - Within the Hubbard-Holstein model, we evaluate the crossover lines marking the opening of pseudogaps in the cuprates, which, in our scenario, are ruled by the proximity to a charge-ordering quantum criticality (stripe formation). We find that their isotopic dependence, due to critical fluctuations, implies a substantial positive shift of the pseudogap-formation temperature T(*). We infer that the isotopic shift of the superconducting T(c) is nearly absent in the optimally and overdoped regimes and is negative and increasing upon underdoping. The dynamical nature of the charge-ordering transition may explain the spread of the experimental values of T(*). PMID- 11497794 TI - Microwave ac conductivity spectrum of a Coulomb glass. AB - We report the first observation of the transition between interacting and noninteracting behavior in the ac conductivity spectrum sigma(omega) of a doped semiconductor in its Coulomb glass state near T = 0 K. The transition manifests itself as a crossover from approximately linear frequency dependence below approximately 10 GHz, to quadratic dependence above approximately 15 GHz. The sharpness of the transition and the magnitude of the crossover frequency strongly suggest that the transition is driven by photon-induced excitations across the Coulomb gap, in contrast to existing theoretical descriptions. PMID- 11497795 TI - Anomalously large critical regions in power-law random matrix ensembles. AB - We investigate numerically the power-law random matrix ensembles. Wave functions are fractal up to a characteristic length whose logarithm diverges asymmetrically with different exponents, 1 in the localized phase and 0.5 in the extended phase. The characteristic length is so anomalously large that for macroscopic samples there exists a finite critical region, in which this length is larger than the system size. The Green's functions decrease with distance as a power law with an exponent related to the correlation dimension. PMID- 11497796 TI - Confinement-enhanced electron transport across a metal-semiconductor interface. AB - We present a combined scanning tunneling microscopy and ballistic electron emission microscopy study of electron transport across an epitaxial Pb/Si(111) interface. Experiments with a self-assembled Pb nanoscale wedge reveal the phenomenon of confinement-enhanced interfacial transport: a proportional increase of the electron injection rate into the semiconductor with the frequency of electron oscillations in the Pb quantum well. PMID- 11497797 TI - Dipolar excitons, spontaneous phase coherence, and superfluid-insulator transition in bilayer quantum Hall systems at nu = 1. AB - The spontaneous interlayer phase coherent (111) state of a bilayer quantum Hall system at filling factor nu = 1 may be viewed as a condensate of interlayer particle-hole pairs or excitons. We show that when the layers are biased in such a way that these excitons are very dilute, they may be viewed as pointlike bosons. We calculate the exciton dispersion relation and show that the exciton exciton interaction is dominated by the dipole moment they carry. In addition to the phase coherent state, we also find a Wigner crystal/glass phase in the presence/absence of disorder which is an insulating state for the excitons. The position of the phase boundary is estimated and the transition between these two phases is discussed. PMID- 11497798 TI - Interband proximity effect and nodes of superconducting gap in Sr2RuO4. AB - The power-law temperature dependences of the specific heat, the nuclear relaxation rate, and the thermal conductivity suggest the presence of line nodes in the superconducting gap of Sr2RuO4. These recent experimental observations contradict the scenario of a nodeless (k(x)+ik(y))-type superconducting order parameter. We propose that interaction of superconducting order parameters on different sheets of the Fermi surface is a key to understanding the above discrepancy. A full gap exists in the active band, which drives the superconducting instability, while line nodes develop in passive bands by the interband proximity effect. PMID- 11497799 TI - Angular position of nodes in the superconducting gap of quasi-2D heavy-fermion superconductor CeCoIn5. AB - The thermal conductivity of the heavy-fermion superconductor CeCoIn5 has been studied in a magnetic field rotating within the 2D planes. A clear fourfold symmetry of the thermal conductivity which is characteristic of a superconducting gap with nodes along the ( +/- pi,+/- pi) directions is resolved. The thermal conductivity measurement also reveals a first-order transition at H(c2), indicating a Pauli limited superconducting state. These results indicate that the symmetry most likely belongs to d(x(2)-y(2)), implying that the anisotropic antiferromagnetic fluctuation is relevant to the superconductivity. PMID- 11497800 TI - Vortex softening: origin of the second peak effect in Bi(2)Sr(2)CaCu(2)O(8+delta). AB - Magnetic hysteresis and transverse ac permeability measurements in Bi(2)Sr(2)CaCu(2)O(8+delta) allow a comparative analysis of the critical current with the elastic response of vortex structures, in equilibrium with their pinning potential, in the field and temperature region where the second peak is detected. This study provides strong evidence that the second peak has its origin in changes of the elastic equilibrium properties of the vortex structures. PMID- 11497801 TI - Bicritical and tetracritical phenomena and scaling properties of the SO(5) theory. AB - By Monte Carlo simulations on a 3D SO(5) rotator model it is shown that, in contrast to the epsilon expansions of the renormalization group, the bicritical point is stable to biquadratic perturbations of AF-SC (antiferromagnetism superconductivity) repulsions, which are produced by quantum fluctuations originated from the Gutzwiller projection. Therefore, the present work completes the link from a physical projected SO(5) starting point to an asymptotic SO(5) symmetry point. The tetracritical point is stable for attractive AF-SC interactions. Critical exponents and ratios are evaluated by scaling analysis. Bicritical and tetracritical scaling functions are derived for the first time. Suggestions on experiments are given. PMID- 11497802 TI - X-ray resonant scattering study of the quadrupolar order in UPd3. AB - Quadrupolar ordering in a 5f electron system has been observed directly for the first time, using x-ray scattering techniques. In UPd (3) at low temperatures satellite peaks appear at (1,0,l) (orthorhombic notation) with l odd and even. Both sets of peaks show a resonant enhancement of the scattering at the M(IV) edge of U. At resonance, the dominant scattering of the l odd peaks occurs in the unrotated polarization channel, whereas for l even a significant rotated component is found. These results are discussed in terms of possible structures of the antiferroquadrupolar phases. PMID- 11497803 TI - Magnetostatic effects in the nucleation of rare earth ferromagnetic phases. AB - It has been reported that superheating, supercooling, and explosive kinetics coupled to other degrees of freedom occur at the ferromagnetic transitions of Er and Dy, and that metastable phases occur during the transition kinetics of Er. We explain these observations in terms of magnetostatic energy, which requires highly eccentric nuclei in the homogeneous nucleation of magnetic transitions in heavy rare earths. The magnetostatics favor transitions through ferrimagnetic intermediaries. The unusual kinetics derive from effective spin lattice relaxation. PMID- 11497804 TI - Nuclear spin driven quantum relaxation in LiY0.998Ho0.002F4. AB - Staircaselike hysteresis loops of the magnetization of a LiY0.998Ho0.002F4 single crystal are observed at subkelvin temperatures and low field sweep rates. This behavior results from quantum dynamics at avoided level crossings of the energy spectrum of single Ho3+ ions in the presence of hyperfine interactions. Enhanced quantum relaxation in constant transverse fields allows the study of the relative magnitude of tunnel splittings. At faster sweep rates, nonequilibrated spin phonon and spin-spin transitions, mediated by weak dipolar interactions, lead to magnetization oscillations and additional steps. PMID- 11497805 TI - Signatures of carrier-wave Rabi flopping in GaAs. AB - For excitation of the model semiconductor GaAs with optical pulses which are both extremely short ( 5 fs) and extremely intense ( approximately 10(12) W cm(-2)), we can meet the condition that the Rabi frequency becomes comparable to the band gap frequency-a highly unusual and previously inaccessible situation. Specifically, in this regime, we observe carrier-wave Rabi flopping, a novel effect of nonlinear optics which has been predicted theoretically and which is related to the failure of the area theorem. PMID- 11497806 TI - Statistical mechanics of prion diseases. AB - We present a two-dimensional, lattice based, protein-level statistical mechanical model for prion diseases (e.g., mad cow disease) with concomitant prion protein misfolding and aggregation. Our studies lead us to the hypothesis that the observed broad incubation time distribution in epidemiological data reflect fluctuation dominated growth seeded by a few nanometer scale aggregates, while much narrower incubation time distributions for innoculated lab animals arise from statistical self-averaging. We model "species barriers" to prion infection and assess a related treatment protocol. PMID- 11497807 TI - Characterization of the branching structure of the lung from "macroscopic" pressure-volume measurements. AB - We analyze the problem of fluid flow in a bifurcating structure containing random blockages that can be removed by fluid pressure. We introduce an asymmetric tree model and find that the predicted pressure-volume relation is connected to the distribution Pi(n) of the generation number n of the tree's terminal segments. We use this relation to explore the branching structure of the lung by analyzing experimental pressure-volume data from dog lungs. The Pi(n) extracted from the data using the model agrees well with experimental data on the branching structure. We can thus obtain information about the asymmetric structure of the lung from macroscopic, noninvasive pressure-volume measurements. PMID- 11497808 TI - Comment on "Observation of superluminal behaviors in wave propagation". PMID- 11497809 TI - Comment on "Observation of superluminal behaviors in wave propagation". PMID- 11497810 TI - Comment on "Repetitive single vortex-loop creation by a vibrating wire in superfluid (3)He-B". PMID- 11497812 TI - Comment on "Phase diagram of an impurity in the spin-1 /2 chain: two-channel Kondo effect versus Curie law". PMID- 11497814 TI - Comment on "Novel convective instabilities in a magnetic fluid". PMID- 11497816 TI - Identification of the radiative and nonradiative parts of a wave field. AB - We present a method for decomposing a wave field, described by a second-order ordinary differential equation, into a radiative component and a nonradiative one, using a biorthonormal system related to the problem under consideration. We show that it is possible to select a special system such that the wave field is purely radiating. We discuss the differences and analogies with approaches which, unlike our approach, start from the corresponding sources of the field. PMID- 11497817 TI - Mesoscopic superpositions of vibronic collective states of N trapped ions. AB - We propose a scalable procedure to generate superpositions of motional coherent states and also entangled vibronic states in N trapped ions. Beyond their fundamental importance, these states may be of interest for quantum information processing and may be used in experimental studies on decoherence. PMID- 11497818 TI - Bose-Einstein condensates with large number of vortices. AB - We show that as the number of vortices in a three dimensional Bose-Einstein condensate increases, the system reaches a "quantum Hall" regime where the density profile is a Gaussian in the xy plane and an inverted parabolic profile along z. The angular momentum of the system increases as the vortex lattice shrinks. However, Coriolis force prevents the unit cell of the vortex lattice from shrinking beyond a minimum size. Although the recent MIT experiment is not exactly in the quantum Hall regime, it is close enough for the present results to be used as a guide. The quantum Hall regime can be easily reached by moderate changes of the current experimental parameters. PMID- 11497819 TI - Constraints on the interaction between dark matter and Baryons from cooling flow clusters. AB - Other nongravitational heating processes are needed to resolve the disagreement between the absence of cool gas components in the centers of galaxy clusters revealed recently by Chandra and XMM observations and the expectations of conventional radiative cooling models. We propose that the interaction between dark matter and baryonic matter may act as an alternative for the reheating of intracluster medium (ICM) in the inner regions of clusters, in which kinetic energy of dark matter is transported to ICM to balance radiative cooling. Using the Chandra and XMM data, we set a useful constraint on the dark-matter-baryon cross section: sigma(xp)/m(x) approximately 1x10(-25) cm(2) GeV-1, where m(x) is the mass of dark matter particles. PMID- 11497820 TI - Localized gravity in string theory. AB - We propose a string realization of the AdS4 brane in AdS5 that is known to localize gravity. Our theory is M D5 branes in the near horizon geometry of N D3 branes, where M and N are appropriately tuned. PMID- 11497823 TI - Asymptotic deconfinement in high-density QCD. AB - We discuss QCD with two light flavors at large baryon chemical potential micro. Color superconductivity leads to partial breaking of the color SU(3) group. We show that the infrared physics is governed by the gluodynamics of the remaining SU(2) group with an exponentially soft confinement scale Lambda(')(QCD) approximately Deltaexp[-a(mu)/(gDelta)], where Delta<< mu is the superconducting gap, g is the strong coupling, and a = 2sqrt[2] pi/11. We estimate that, at moderate baryon densities, Lambda(')(QCD) is O(10 MeV) or smaller. The confinement radius increases exponentially with density, leading to "asymptotic deconfinement." The velocity of the SU(2) gluons is small due to the large dielectric constant of the medium. PMID- 11497824 TI - Random versus realistic interactions for low-lying nuclear spectra. AB - We compare the shell-model results for realistic interactions with those obtained for various ensembles of random matrix elements. We show that, although the quantum numbers of the ground states in the even-even nuclei have a high probability ( approximately 60%) to be J(pi)T = 0(+)0, the overlap of those states with the realistic wave functions is very small in average. The transition probabilities B(E2) predicted with random interactions are also too small. The presence of the regular pairing is shown to be a significant element of realistic physics not reproduced by random interactions. PMID- 11497821 TI - Experimental investigation of the two-photon widths of the chi(c0) and the chi(c2) mesons. AB - Using 12.7 fb(-1) of data collected with the CLEO detector at CESR, we observed two-photon production of the cc states chi(c0) and chi(c2) in their decay to pi(+)pi(-)pi(+)pi(-). We measured gamma(gammagamma)(chi(c))xB(chi(c)-->pi(+)pi( )pi(+)pi(-)) to be 75+/-13(stat)+/-8(syst) eV for the chi(c0) and 6.4+/ 1.8(stat)+/-0.8(syst) eV for the chi(c2), implying gamma(gammagamma)(chi(c0)) = 3.76+/-0.65(stat)+/-0.41(syst)+/-1.69(br) keV and gamma(gammagamma)(chi(c2)) = 0.53+/-0.15(stat)+/-0.06(syst)+/-0.22(br) keV. Also, cancellation of dominant experimental and theoretical uncertainties permits a precise comparison of gamma(gammagamma)(chi(c0))/gamma(gammagamma)(chi(c2)), evaluated to be 7.4+/ 2.4(stat)+/-0.5(syst)+/-0.9(br), with QCD-based predictions. PMID- 11497825 TI - Magic nucleus 132Sn and its one-neutron-hole neighbor 131Sn. AB - Prompt and delayed gamma-ray cascades in doubly magic 132Sn and its neighbor 131Sn have been studied at Gammasphere using a 248Cm fission source. Isotopic assignments of unknown gamma rays were based on coincidences with known transitions in A = 112-116 Pd fission partners. The yrast level spectra of both tin nuclei are interpreted using empirical nucleon-nucleon interactions from the 132Sn and 208Pb regions. Results include identification of the (nuf(7/2)h( 1)(11/2))9(+) aligned state in 132Sn and of extensive (nuf(7/2)h(-2)(11/2)), (nuf(7/2)d(-1)(3/2)h(-1)(11/2)) and (nuh(-1)(11/2)x3(-)) multiplets in 131Sn. The previously reported beta(-) decay of an unusual 131In high-spin isomer to levels in 131Sn is also elucidated. PMID- 11497826 TI - Nuclear quadrupole moment of 57Fe from microscopic nuclear and atomic calculations. AB - The nuclear quadrupole moment (NQM) of the Ipi = 3/2(-) excited nuclear state of 57Fe at 14.41 keV, important in Mossbauer spectroscopy, is determined from the large-scale nuclear shell-model calculations for 54Fe, 57Fe, and also from the electronic ab initio and density functional theory calculations including solid state and electron correlation effects for the molecules Fe(CO)(5) and Fe(C5H5)(2). Both independent methods yield very similar results. The recommended value is 0.15(2) e b. The NQM of the isomeric 10+ in 54Fe has also been calculated. The new NQM values for 54Fe and 57Fe are consistent with the perturbed angular distribution data. PMID- 11497827 TI - Elastic positronium-atom scattering using the stochastic variational method. AB - The stochastic variational method is used in conjunction with stabilization ideas to compute the low energy phase shifts and scattering lengths for positronium atom scattering. Results are obtained for the Ps-H, Ps- Li+, Ps-He, and Ps-Ps systems. The Ps-H scattering lengths are probably accurate to better than 5% and are the most accurate so far computed. The results for Ps- Li+ and Ps-Ps scattering represent the first published scattering lengths for these systems. The positive scattering length for completely spin-aligned 3Ps-3Ps scattering, namely 2.95a(0), is particularly significant since it demonstrates the feasibility of forming a stable Bose-Einstein condensate of 3Ps atoms. PMID- 11497828 TI - Charge-induced fragmentation of sodium clusters. AB - The fission of highly charged sodium clusters with fissilities X>1 is studied by ab initio molecular dynamics. Na4+24 is found to undergo predominantly sequential Na+3 emission on a time scale of 1 ps, while Na(Q+)(24) ( 5< or =Q< or =8) undergoes multifragmentation on a time scale > or =0.1 ps, with Na+ increasingly the dominant fragment as Q increases. All singly charged fragments Na(+)(n) up to size n = 6 are observed. The observed fragment spectrum is, within statistical error, independent of the temperature T of the parent cluster for T< or =1500 K. These findings are consistent with and explain recent trends observed experimentally. PMID- 11497829 TI - Resonance-assisted tunneling in near-integrable systems. AB - Dynamical tunneling between symmetry related invariant tori is studied in the near-integrable regime. Using the kicked Harper model as an illustration, we show that the exponential decay of the wave functions in the classically forbidden region is modified due to coupling processes that are mediated by classical resonances. This mechanism leads to a substantial deviation of the splitting between quasidegenerate eigenvalues from the purely exponential decrease with 1/Planck's over 2pi obtained for the integrable system. A simple semiclassical framework, which takes into account the effect of the resonance substructure on the invariant tori, allows one to quantitatively reproduce the behavior of the eigenvalue splittings. PMID- 11497830 TI - Scaling properties of the temperature field in convective turbulence. AB - We report the scaling properties of temperature in turbulent convection in water. In the central region of the convection cell, we find that the peak frequency of the temperature dissipation spectra may be identified as the "Bolgiano frequency," with respect to which the temperature power spectra are universal functions; and that the usual inertial range is taken up entirely by the buoyancy subrange, so that a "high frequency" scaling subrange emerges only through an extended-self-similarity-type analysis. Moreover, the buoyancy subrange assumes the value of 2/5 predicted for the Bolgiano-Obukhov scaling only in the central region of the cell; in the mixing zone the exponent for the high frequency scaling exponent has a value of 2/3. PMID- 11497831 TI - Slow group velocity and Cherenkov radiation. AB - We study theoretically the effect of ultraslow group velocities on the emission of Vavilov-Cherenkov radiation in a coherently driven medium. We show that in this case the aperture of the group cone on which the intensity of the radiation peaks is much smaller than that of the usual wave cone associated with the Cherenkov coherence condition. As a specific example, we consider a coherently driven ultracold atomic gas where such singular behavior may be observed. PMID- 11497822 TI - Search for heavy particles decaying into electron-positron pairs in pp collisions. AB - We present results of searches for technirho (rho(T)), techniomega (omega(T)), and Z' particles, using the decay channels rho(T),omega(T),Z'-->e(+)e(-). The search is based on 124.8 pb(-1) of data collected by the D0 detector at the Fermilab Tevatron during 1992-1996. In the absence of a signal, we set 95% C.L. upper limits on the cross sections for the processes pp-->rho(T),omega(T),Z'- >e(+)e(-) as a function of the mass of the decaying particle. For certain model parameters, we exclude the existence of degenerate rho(T) and omega(T) states with masses below about 200 GeV. We exclude a Z' with mass below 670 GeV, assuming that it has the same couplings to fermions as the Z boson. PMID- 11497832 TI - Characterization of stationary and nonstationary behavior in gyrotron oscillators. AB - The transition from the stationary state to a sequence of nonstationary states in the gyromonotron oscillator is experimentally characterized for the first time. We have also demonstrated the stationary operation of a gyrotron backward-wave oscillator at a beam current far in excess of the generally predicted nonstationary threshold. This difference in nonlinear behavior has been investigated and shown to be fundamental with a comparative analysis of the feedback mechanisms, energy deposition profiles, and field shaping processes involved in these two types of oscillations. PMID- 11497833 TI - Experimental evidence of intermittent convection in the edge of magnetic confinement devices. AB - Probe measurements in the PISCES linear device indicate the presence of plasma radially far from where it is produced. We show that this is mainly caused by large-scale structures of plasma with high radial velocity. Data from the Tore Supra tokamak show striking similarities in the shape of these intermittent events as well as the fluctuation density probability distribution and frequency spectrum. The fact that intermittent, large-scale events are so similar in linear devices and tokamaks indicates the universality of convective transport in magnetically confined plasmas. PMID- 11497834 TI - Microscopic kinetics and time-dependent structure factors. AB - The time evolution of structure factors (SF) in the disordering process of an initially phase-separated lattice depends crucially on the microscopic disordering mechanism, such as Kawasaki dynamics (KD) or vacancy-mediated disordering (VMD). Monte Carlo simulations show unexpected "dips" in the SFs. A phenomenological model is introduced to explain the dips in the odd SFs, and an analytical solution of KD is derived, in excellent agreement with simulations. The presence (absence) of dips in the even SFs for VMD (KD) marks a significant but not yet understood difference of the two dynamics. PMID- 11497835 TI - Bulk and boundary zero-bias anomaly in multiwall carbon nanotubes. AB - We compute the tunneling density of states of doped multiwall nanotubes including disorder and electron-electron interactions. A nonconventional Coulomb blockade reflecting nonperturbative Altshuler-Aronov-Lee power-law zero-bias anomalies is found, in accordance with recent experimental results. The presence of a boundary implies a universal doubling of the boundary exponent in the diffusive limit. PMID- 11497836 TI - Glassy spin dynamics in non-fermi-liquid UCu5-xPdx, x = 1.0 and 1.5. AB - Local f-electron spin dynamics in the non-Fermi-liquid heavy-fermion alloys UCu5 xPdx, x = 1.0 and 1.5, have been studied using muon spin-lattice relaxation. The sample-averaged asymmetry function G(t) indicates strongly inhomogeneous spin fluctuations and exhibits the scaling G(t,H) = G(t/H(gamma)) expected from glassy dynamics. At 0.05 K gamma(x = 1.0) = 0.35+/-0.1, but gamma(x = 1.5) = 0.7+/-0.1. This is in contrast to inelastic neutron scattering results, which yield gamma = 0.33 for both concentrations. There is no sign of static magnetism approximately greater than 10(-3)(B)/U ion in either material above 0.05 K. Our results strongly suggest that both alloys are quantum spin glasses. PMID- 11497837 TI - Class of exactly solvable pairing models. AB - We present three classes of exactly solvable models for fermion and boson systems, based on the pairing interaction. These models are solvable in any dimension. As an example we show the first results for fermions interacting with repulsive pairing forces in a two-dimensional square lattice. In spite of the repulsive pairing force the exact results show attractive pair correlations. PMID- 11497838 TI - Spin, charge, and orbital ordering in La0.5Sr1.5Mno4. AB - We have analyzed the experimental evidence of charge and orbital ordering in La0.5Sr1.5MnO4 using first principles band structure calculations. Our results suggest the presence of two types of Mn sites in the system. One of the Mn sites behaves as an Mn3+ ion, favoring a Jahn-Teller distortion of the surrounding oxygen atoms, while the distortion around the other is not a simple breathing mode kind. Band structure effects are found to dominate the experimental spectrum for orbital and charge ordering, providing an alternate explanation for the experimentally observed results. PMID- 11497839 TI - Dimer decimation and intricately nested localized-ballistic phases of a kicked Harper model. AB - A new decimation scheme is introduced to study localization transitions in tight binding models with long range interaction. Within this scheme, the lattice models are mapped to a vectorized dimer where an asymptotic dissociation of the dimer is shown to correspond to the vanishing of the transmission coefficient through the system. When applied to the kicked Harper model, the method unveils an intricately nested extended and localized phases in two-dimensional parameter space. In addition to computing transport characteristics with extremely high precision, the renormalization tools also provide a new method to compute quasienergy spectrum. PMID- 11497840 TI - Dissociation of vertical semiconductor diatomic artificial molecules. AB - We investigate the dissociation of few-electron circular vertical semiconductor double quantum dot artificial molecules at 0 T as a function of interdot distance. A slight mismatch introduced in the fabrication of the artificial molecules from nominally identical constituent quantum wells induces localization by offsetting the energy levels in the quantum dots by up to 2 meV, and this plays a crucial role in the appearance of the addition energy spectra as a function of coupling strength particularly in the weak coupling limit. PMID- 11497841 TI - Rectification properties of carbon nanotube "Y-junctions". AB - Quantum conductivity of single-wall carbon nanotube Y-junctions is calculated. The current versus voltage characteristics of these junctions show asymmetry and rectification, in agreement with recent experimental results. Furthermore, rectification is found to be independent of the angle between the branches of these junctions, indicating this to be an intrinsic property of symmetric Y junctions. The implications for the Y-junction to function as a nanoscale molecular electronic switch are investigated. PMID- 11497842 TI - Spin injection across a heterojunction: a ballistic picture. AB - Spin injection across heterojunctions plays a decisive role in the new field of spintronics. Within the ballistic transport regime, we state a general expression for the spin-injection rate in a heterojunction made of two ballistic electrodes. Both the spin-orbit interaction and interface scattering effect are taken into account. Our model is consistent with the well-documented results of ferromagnetic-metal junctions. It explains the recent experimental results of a dilute-magnetic-semiconductor/semiconductor junction and predicts solutions to enhance the spin-injection rate across a ferromagnetic-semiconductor junction. PMID- 11497843 TI - Anomalous finite-size effect in superconducting Josephson junction arrays. AB - We show that a previously reported discrepancy between simulations of superconducting Josephson junction arrays and the theoretical analysis of Ambegaokar, Halperin, Nelson, and Siggia (AHNS) [Phys. Rev. Lett. 40, 783 (1978)] is rooted in a peculiar finite-size effect under periodic boundary conditions. Our simulation results for the largest array support the power-law I-V curves predicted by AHNS. Analysis of the vortex dynamics reveals two intrinsic length scales set by the applied current, which define three size regimes with distinctive I-V characteristics. PMID- 11497844 TI - Superconductivity in an organic insulator at very high magnetic fields. AB - We investigate by electrical transport the field-induced superconducting state (FISC) in the organic conductor lambda-(BETS)2FeCl4. Below 4 K, antiferromagnetic insulator, metallic, and eventually superconducting (FISC) ground states are observed with increasing in-plane magnetic field. The FISC state survives between 18 and 41 T and can be interpreted in terms of the Jaccarino-Peter effect, where the external magnetic field compensates the exchange field of aligned Fe3+ ions. We further argue that the Fe3+ moments are essential to stabilize the resulting singlet, two-dimensional superconducting state. PMID- 11497845 TI - Vortex avalanches and magnetic flux fragmentation in superconductors. AB - We report the results of numerical simulations of nonisothermal dendritic flux penetration in type-II superconductors. We propose a generic mechanism of dynamic branching of a propagating hot spot of a flux flow/normal state triggered by a local heat pulse. The branching occurs when the flux hot spot reflects from inhomogeneities or the boundary on which magnetization currents either vanish, or change direction. The hot spot then undergoes a cascade of successive splittings, giving rise to a dissipative dendritic-type flux structure. This dynamic state eventually cools down, turning into a frozen multifilamentary pattern of magnetization currents. PMID- 11497846 TI - Time-resolved measurements of thermodynamic fluctuations of the particle number in a nondegenerate Fermi gas. AB - We report on time-resolved measurements of thermodynamic fluctuations in the number of particles in a nondegenerate Fermi gas. The gas is comprised of thermal quasiparticles, confined in a superconducting Al box by large-gap Ta leads. The average number of quasiparticles is about 10(5). This number fluctuates due to quasiparticle generation and recombination. The number is measured from the tunneling current through a barrier that bisects the box. The recombination time is independently measured by single-photon excitation and agrees with the frequency dependence of the fluctuations. PMID- 11497847 TI - Correlation of tunneling spectra in Bi(2)Sr(2)CaCu(2)O(8+delta) with the resonance spin excitation. AB - New break-junction tunneling data are reported in Bi(2)Sr(2)CaCu(2)O(8+delta) over a wide range of hole concentration from underdoped (T(c) = 74 K) to optimal doped (T(c) = 95 K) to overdoped (T(c) = 48 K). The conductances exhibit sharp dips at a voltage, Omega/e, measured with respect to the superconducting gap. Clear trends are found such that the dip strength is maximum at optimal doping and that Omega scales as 4.9kT(c) over the entire doping range. These features link the dip to the resonance spin excitation and suggest quasiparticle interactions with this mode are important for superconductivity. PMID- 11497848 TI - Full current statistics in diffusive normal-superconductor structures. AB - We study the current statistics in normal diffusive conductors in contact with a superconductor. Using an extension of the Keldysh Green's function method we are able to find the full distribution of charge transfers for all temperatures and voltages. For the non-Gaussian regime, we show that the equilibrium current fluctuations are enhanced by the presence of the superconductor. We predict an enhancement of the nonequilibrium current noise for temperatures below and voltages of the order of the Thouless energy E(Th) = D/L(2). Our calculation fully accounts for the proximity effect in the normal metal and agrees with experimental data. PMID- 11497849 TI - Do superconductors have zero resistance in a magnetic field? AB - We show that dc voltage versus current measurements of a YBa(2)Cu(3)O(7-delta) film in a magnetic field can be collapsed onto scaling functions proposed by Fisher et al. [Phys. Rev. B 43, 130 (1991)] as is widely reported in the literature. We find, however, that good data collapse is achieved for a wide range of critical exponents and temperatures. These results strongly suggest that agreement with scaling alone does not prove the existence of a phase transition. We propose a criterion to determine if the data collapse is valid, and thus if a phase transition occurs. To our knowledge, none of the data reported in the literature meet our criterion. PMID- 11497850 TI - Anisotropic spin-orbit coupling and magnetocrystalline anisotropy in vicinal co films. AB - The anisotropy of the spin-orbit interaction, , in vicinal Co films has been measured using x-ray magnetic linear dichroism (XMLD). A linear increase in with Co step density is found using a new sum rule and represents the first experimental confirmation that XMLD probes the magnetocrystalline anisotropy energy (MAE). X-ray magnetic circular dichroism is used to confirm that the XMLD arises from changes in the local step-edge electronic structure. The XMLD sum rule gives a larger MAE compared to macroscopic values and is discussed with respect to other local probes of the MAE. PMID- 11497851 TI - Spin-ordering quantum transitions of superconductors in a magnetic field. AB - We argue that recent neutron scattering measurements by Lake et al. [Science 291, 1759 (2001)] of the spin excitation spectrum of La(2-delta)Sr(delta)CuO4 in a magnetic field can be understood in terms of proximity to a phase with co existing superconductivity and spin density wave order. We present a general theory for such quantum transitions, and argue that their low energy spin fluctuations are controlled by a singular correction from the superflow kinetic energy, acting in the region outside the vortex cores. We propose numerous experimental tests of our theory. PMID- 11497852 TI - Long-range order at low temperatures in dipolar spin ice. AB - It has recently been suggested that long-range magnetic dipolar interactions are responsible for spin ice behavior in the Ising pyrochlore magnets Dy2Ti2O7 and Ho2Ti2O7. We report here numerical results on the low temperature properties of the dipolar spin ice model, obtained via a new loop algorithm which greatly improves the dynamics at low temperature. We recover the previously reported missing entropy in this model, and find a first order transition to a long-range ordered phase with zero total magnetization at very low temperature. We discuss the relevance of these results to Dy2Ti2O7 and Ho2Ti2O7. PMID- 11497853 TI - Quantum decoherence in the spectral functions of undoped lamno3. AB - We study the one hole spectral function in a model for LaMnO3 including both the effects of orbital ordering and the quantum decoherence due to the antiferromagnetic coupling between the ferromagnetic layers. We find that the classical picture of a ferromagnetic polaron does not apply and free dispersion is replaced by rigid quasiparticles on the scale of magnetic excitations, while the spectra are dominated by the incoherent spectral weight at high energies. These results have important implications on the in-plane transport and angular resolved photoemission in the manganites. PMID- 11497854 TI - Finite-temperature magnetism of transition metals: an ab initio dynamical mean field theory. AB - We present an ab initio quantum theory of the finite-temperature magnetism of iron and nickel. A recently developed technique which combines dynamical mean field theory with realistic electronic structure methods successfully describes the many-body features of the one electron spectra and the observed magnetic moments below and above the Curie temperature. PMID- 11497855 TI - Hybridized nature of pseudogap in Kondo insulators CeRhSb and CeRhAs. AB - We studied the electronic structure of Kondo insulators CeRhSb and CeRhAs using high-resolution photoemission spectroscopy. We found that the 4f-derived density of states shows a depletion (pseudogap) at E(F) in contrast to metallic Kondo materials. It was found that the size of the f pseudogap is smaller than that of conduction electrons ( c pseudogap) while both scale well with the Kondo temperature. The present results indicate that the hybridization between 4f and conduction electrons near E(F) is essential for the Kondo gap in the Ce-based compounds. PMID- 11497856 TI - Selective-resputtering-induced perpendicular magnetic anisotropy in amorphous TbFe films. AB - Perpendicular magnetic anisotropy energy in rf magnetron sputtered amorphous TbFe films is measured to increase exponentially with pair-order anisotropy induced by the selective resputtering of surface adatoms during film growth. PMID- 11497857 TI - Realization of a large magnetic moment in the ferromagnetic CoPt bulk phase. AB - Changes in the magnetic moment and other physical properties of a CoPt alloy induced by a new type of ion-beam mixing in an external magnetic field were investigated. This process induces the formation of a metastable phase through extremely rapid quenching from well above the ordering temperature. The measured magnetic moment per Co atom was 2.63 mu(B), larger by 55% and 35% than that of the bulk Co and stable CoPt film, respectively, which is one of the highest values ever observed in the ferromagnetic bulk phase. PMID- 11497858 TI - Long-range order and low-energy spectrum of diluted 2D quantum antiferromagnet. AB - The problem of a diluted two-dimensional quantum antiferromagnet on a square lattice is studied using spin-wave theory. The influence of impurities on static and dynamic properties is investigated and a good agreement with experiments and Monte Carlo data is found. The hydrodynamic description of spin waves breaks down at characteristic wavelengths Lambda greater than exp(const/x), x being an impurity concentration, while the order parameter is free from anomalies. We argue that this dichotomy originates from strong scattering of the low-energy excitations in two dimensions. PMID- 11497859 TI - Nanoscale multiphase separation at La(2/3)Ca(1/3)MnO3/SrTiO3 interfaces. AB - 55Mn nuclear magnetic resonance experiments are reported on a series of fully strained epitaxial La(2/3)Ca(1/3)MnO3 thin films on SrTiO3. We have found evidence of multiple phase segregation into ferromagnetic metallic and nonmetallic regions as well as regions that are nonferromagnetic and insulating. These insulating regions are mainly located close to interfaces and may have a significant impact on the performance of spin-tunnel devices. As a result of phase segregation, the ferromagnetic coupling within the metallic regions is depressed. This accounts for the reduction of the Curie temperature and conductivity in nanometric thin films. PMID- 11497860 TI - Control of exciton dynamics in nanodots for quantum operations. AB - We present a theory to further a new perspective of proactive control of exciton dynamics in the quantum limit. Circularly polarized optical pulses in a semiconductor nanodot are used to control the dynamics of two interacting excitons of opposite polarizations. Shaping of femtosecond laser pulses keeps the quantum operation within the decoherence time. Computation of the fidelity of the operations and application to the complete solution of a minimal quantum computing algorithm demonstrate in theory the feasibility of quantum control. PMID- 11497861 TI - Lifetime measurements with a scanning positron microscope. AB - First lifetime results obtained with a scanning positron microscope will be presented. A pulsed positron beam with a variable energy from 0.5 to 20 keV, with a spot diameter of 2 microm, can be electronically scanned over an area of 0.6x0.6 mm(2). This beam is formed after a double-stage stochastic cooling (moderation) of positrons emitted from a radioactive isotope. Included in the system is a conventional scanning electron microprobe for surface analysis. Three dimensional positron lifetime spectra of a GaAs sample with a small surface scratch reveal the range due to the mechanical damage. PMID- 11497862 TI - Transverse spin relaxation in liquid 129Xe in the presence of large dipolar fields. AB - Using spin-echo NMR techniques we study the transverse spin relaxation of hyperpolarized liquid 129Xe in a spherical cell. We observe an instability of the transverse magnetization due to dipolar fields produced by liquid 129Xe, and find that imperfections in the pi pulses of the spin-echo sequence suppress this instability. A simple perturbative model of this effect is in good agreement with the data. We obtain a transverse spin relaxation time of 1300 sec in liquid 129Xe, and discuss applications of hyperpolarized liquid 129Xe as a sensitive magnetic gradiometer and for a permanent electric dipole moment search. PMID- 11497863 TI - Probabilistic quantum memories. AB - Typical address-oriented computer memories cannot recognize incomplete or noisy information. Associative (content-addressable) memories solve this problem but suffer from severe capacity shortages. I propose a model of a quantum memory that solves both problems. The storage capacity is exponential in the number of qbits and thus optimal. The retrieval mechanism for incomplete or noisy inputs is probabilistic, with postselection of the measurement result. The output is determined by a probability distribution on the memory which is peaked around the stored patterns closest in Hamming distance to the input. PMID- 11497864 TI - Resetting wave forms in dictyostelium territories. AB - The mechanism by which spiral wave patterns appear in populations of Dictyostelium was probed experimentally by external chemical perturbation. Spiral waves, which often arise from the breakup of circular waves driven by pacemakers, typically entrain those pacemakers. We studied these processes by resetting the waves with a spatially uniform pulse of extrinsic cyclic AMP. A pattern of spirals reappeared if resetting was early in the signaling stage, but only targets emerged following late resetting, in a manner analogous to cardiac defibrillation. This supports recent hypotheses that wave pattern selection naturally occurs by slow temporal variation of the excitability of the cells. PMID- 11497865 TI - Dynamical encoding by networks of competing neuron groups: winnerless competition. AB - Following studies of olfactory processing in insects and fish, we investigate neural networks whose dynamics in phase space is represented by orbits near the heteroclinic connections between saddle regions (fixed points or limit cycles). These networks encode input information as trajectories along the heteroclinic connections. If there are N neurons in the network, the capacity is approximately e(N-1)!, i.e., much larger than that of most traditional network structures. We show that a small winnerless competition network composed of FitzHugh-Nagumo spiking neurons efficiently transforms input information into a spatiotemporal output. PMID- 11497866 TI - The future is noisy: the role of spatial fluctuations in genetic switching. AB - A genetic switch may be realized by a certain operator sector on the DNA strand from which either genetic code, to the left or to the right of this operator sector, can be transcribed and the corresponding information processed. This switch is controlled by messenger molecules, i.e., they determine to which side the switch is flipped. Recently, it has been realized that noise plays an elementary role in genetic switching, and the effect of number fluctuations of the messenger molecules have been explored. Here we argue that the assumption of well stirredness taken in the previous models may not be sufficient to characterize the influence of noise: spatial fluctuations play a non-negligible part in cellular genetic switching processes. PMID- 11497868 TI - Strong explosive interaction of hydrogenated porous silicon with oxygen at cryogenic temperatures. AB - We report new types of heterogeneous hydrogen-oxygen and silicon-oxygen branched chain reactions which have been found to proceed explosively after the filling of pores of hydrogen-terminated porous silicon (Si) by condensed or liquid oxygen in the temperature range of 4.2-90 K. Infrared vibrational absorption spectroscopy shows that, while initially Si nanocrystals assembling the layers have hydrogen terminated surfaces, the final products of the reaction are SiO2 and H2O. Time resolved optical experiments show that the explosive reaction develops in a time scale of 10(-6) s. We emphasize the remarkable structural properties of porous Si layers which are crucial for the strong explosive interaction. PMID- 11497867 TI - Noise effects on the complex patterns of abnormal heartbeats. AB - Patients at high risk for sudden death often exhibit complex heart rhythms in which abnormal heartbeats are interspersed with normal heartbeats. We analyze such a complex rhythm in a single patient over a 12-h period and show that the rhythm can be described by a theoretical model consisting of two interacting oscillators with stochastic elements. By varying the magnitude of the noise, we show that for an intermediate level of noise, the model gives best agreement with key statistical features of the dynamics. PMID- 11497869 TI - Communication networks with an emergent dynamical structure. AB - A model with a dynamical network structure is studied. The essential difference from other models is that a disappearance of links is also allowed. The obtained results suggest that under certain conditions a fairly robust cluster, which contains all of the elements of the system, can be formed. According to the power law dependence in the node connectivity distribution, a scale-free regime can occur in such models. The average network diameter behaves as K-tau. The emergent structure is close to the small-world networks for certain values of p and to the large-world ones for others. Generally, the influence of the parameter K on the average network diameter is more significant than that of the system size. PMID- 11497870 TI - Comment on "can disorder induce a finite thermal conductivity in 1D lattices?". PMID- 11497872 TI - Comment on "quantum games and quantum strategies". PMID- 11497874 TI - Off-diagonal geometric phase in a neutron interferometer experiment. AB - Off-diagonal geometric phases acquired by an evolution of a 1/2-spin system have been observed by means of a polarized neutron interferometer. We have successfully measured the off-diagonal phase for noncyclic evolutions even when the diagonal geometric phase is undefined. Our data confirm theoretical predictions and the results illustrate the significance of the off-diagonal phase. PMID- 11497875 TI - Classical and quantum Hamiltonian ratchets. AB - We explain the mechanism leading to directed chaotic transport in Hamiltonian systems with spatial and temporal periodicity. We show that a mixed phase space comprising both regular and chaotic motion is required and we derive a classical sum rule which allows one to predict the chaotic transport velocity from properties of regular phase-space components. Transport in quantum Hamiltonian ratchets arises by the same mechanism as long as uncertainty allows one to resolve the classical phase-space structure. We derive a quantum sum rule analogous to the classical one, based on the relation between quantum transport and band structure. PMID- 11497876 TI - Peculiar scaling of self-avoiding walk contacts. AB - The nearest neighbor contacts between the two halves of an N-site lattice self avoiding walk offer an unusual example of scaling random geometry: for N- >infinity they are strictly finite in number but their radius of gyration R(c) is power law distributed proportional to R(-tau)(c), where tau>1 is a novel exponent characterizing universal behavior. A continuum of diverging length scales is associated with the R(c) distribution. A possibly superuniversal tau = 2 is also expected for the contacts of a self-avoiding or random walk with a confining wall. PMID- 11497877 TI - TeV neutrinos and GeV photons from shock breakout in supernovae. AB - We show that as a Type II supernova shock breaks out of its progenitor star, it becomes collisionless and may accelerate protons to energies >10 TeV. Inelastic nuclear collisions of these protons produce an approximately 1 h long flash of TeV neutrinos and 10 GeV photons, about 10 h after the thermal (10 MeV) neutrino burst from the cooling neutron star. A Galactic supernova in a red supergiant star would produce a photon and neutrino flux of approximately 10(-4) erg cm(-2) s(-1). A km(2) neutrino detector will detect approximately 100 muons, thus allowing to constrain both supernova models and neutrino properties. PMID- 11497879 TI - Measurement of the branching ratio and form factor of K(L)-->mu(+)mu(-)gamma. AB - We report on the analysis of the rare decay K(L)-->mu(+)mu(-)gamma the 1997 data from the KTeV experiment at Fermilab. A total of 9327 candidate events are observed with 2.4% background, representing a factor of 40 increase in statistics over the current world sample. We find that B(K(L)-->mu(+)mu(-)gamma) = (3.62 +/- 0.04(stat) +/- 0.08(syst)) x 10(-7). The form factor parameter alpha(K*) is measured to be alpha(K*) = -0.160(+0.026)(-0.028). In addition, we make the first measurement of the parameter alpha from the D'Ambrosio-Isidori-Portoles form factor, finding alpha = -1.54 +/- 0.10. In that model, this alpha measurement limits the Cabibbo-Kobayashi-Maskawa parameter rho>-0.2. PMID- 11497881 TI - Search for the lepton family number violating process nu(mu)e(-) --> mu(-)nu(e). AB - The NuTeV experiment at Fermilab has used a sign-selected neutrino beam to perform a search for the lepton number violating process nu(mu)e(-)-->mu(-)nu(e), and to measure the cross section of the standard model inverse muon decay process nu(mu)e(-)-->mu(-)nu(e). NuTeV measures the inverse muon decay asymptotic cross section slope sigma/E to be (13.8 +/- 1.2 +/- 1.4) x 10(-42) cm(2)/GeV. The experiment also observes no evidence for lepton number violation and places one of the most restrictive limits on the cross-section ratio sigma(nu(mu)e(-)-->mu( )nu(e))/sigma(nu(mu)e(-)-->mu(-)nu(e)) < or = 1.7% at 90% C.L. for V-A couplings and < or = 0.6% for scalar couplings. PMID- 11497878 TI - Measurement of the rate of nu(e) + d --> p + p + e(-) interactions produced by (8)B solar neutrinos at the Sudbury Neutrino Observatory. AB - Solar neutrinos from (8)B decay have been detected at the Sudbury Neutrino Observatory via the charged current (CC) reaction on deuterium and the elastic scattering (ES) of electrons. The flux of nu(e)'s is measured by the CC reaction rate to be straight phi(CC)(nu(e)) = 1.75 +/- 0.07(stat)(+0.12)(-0.11)(syst) +/- 0.05(theor) x 10(6) cm(-2) s(-1). Comparison of straight phi(CC)(nu(e)) to the Super-Kamiokande Collaboration's precision value of the flux inferred from the ES reaction yields a 3.3 sigma difference, assuming the systematic uncertainties are normally distributed, providing evidence of an active non- nu(e) component in the solar flux. The total flux of active 8B neutrinos is determined to be 5.44+/-0.99 x 10(6) cm(-2) s(-1). PMID- 11497882 TI - Hadronic loop corrections to the muon anomalous magnetic moment. AB - The dominant theoretical uncertainties in both the anomalous magnetic moment of the muon and the value of the electromagnetic coupling at the Z scale, M(Z), arise from their hadronic contributions. Since these will ultimately dominate the experimental errors, we study the correlation between them, as well as with other fundamental parameters. To this end we present analytical formulas for the QCD contribution from higher energies and from heavy quarks. Including these correlations affects the Higgs boson mass extracted from precision data. PMID- 11497883 TI - Model-independent analysis of muon g(mu) - 2. AB - We consider possible physics beyond the standard model that could account for the large recently reported [Muon g-2 Collaboration, H. N. Brown et al., Phys. Rev. Lett. 86, 2227 (2001)] effect in g(mu)-2. If the underlying theory can be treated perturbatively, then the only possibilities are models that contain particles that yield "direct" contributions, e.g., supersymmetric models, models with unexpectedly light leptoquarks, or with a rather exotic heavy vector boson. If the underlying theory involves strong interactions, as in technicolor models, then the discrepancy could be accounted for by a variety of mechanisms. PMID- 11497884 TI - Chiral extrapolation, renormalization, and the viability of the quark model. AB - The relationship of the quark model to the known chiral properties of QCD is a long-standing problem in the interpretation of low-energy QCD. In particular, how can the pion be viewed as both a collective Goldstone-boson quasiparticle and as a valence-quark-antiquark bound state? A comparison of the many-body solution of a simplified model of QCD to the constituent-quark model demonstrates that the quark model is sufficiently flexible to describe meson hyperfine splitting provided proper renormalization conditions and correct degrees of freedom are employed consistently. PMID- 11497885 TI - Isomer spectroscopy in (216)(90)Th(126) and the magicity of (218)(92)U(126). AB - Excited states in (216)Th were investigated via prompt and delayed gamma decays and the recoil-decay tagging method. The decay schemes of the I(pi) = (8+), t(1/2) = 128(8) micros, the I(pi) = (11-), t(1/2) = 615(55) ns, and the I(pi) = (14+), t(1/2) > or = 130 ns isomers were established. The configuration pi h(9/2)f(7/2) is assigned to the I(pi) = (8+) isomer, which implies that the h(9/2) and f(7/2) states are nearly degenerate. This is ascribed to increased binding of the f(7/2) orbital by its coupling to a low-lying I(pi) = (3-) state at E(x) = 1687 keV. The role of octupole and pairing correlations for a Z = 92 shell closure prediction is discussed on the basis of shell model calculations. PMID- 11497886 TI - Test of nuclear wave functions for pseudospin symmetry. AB - Using the fact that pseudospin is an approximate symmetry of the Dirac Hamiltonian with realistic scalar and vector mean fields, we derive the wave functions of the pseudospin partners of eigenstates of a realistic Dirac Hamiltonian and compare these wave functions with the wave functions of the Dirac eigenstates. PMID- 11497880 TI - Rate measurement of D(0)-->K+pi(-)pi(0) and constraints on D(0) -- D(0) mixing. AB - We present an observation and time-integrated rate measurement of the decay D(0)- >K(+)pi(-)pi(0) produced in 9 fb(-1) of e(+)e(-) collisions near the Upsilon(4S) resonance. The signal is inconsistent with an upward fluctuation of the background by 4.9 standard deviations. We measured the time-integrated rate of D(0)-->K(+)pi(-)pi(0) normalized to the rate of D(0)-->K(+)pi(-)pi(0) to be 0.0043(+0.0011)(-0.0010) (stat)+/-0.0007 (syst). This decay can be produced by doubly Cabibbo-suppressed decays or by the D(0) evolving into a D(0) through mixing, followed by a Cabibbo-favored decay to K(+)pi(-)pi(0). We also found the CP asymmetry A = (9(+25)(-22))% be consistent with zero. PMID- 11497887 TI - Search for x-ray induced acceleration of the decay of the 31-yr isomer of (178)Hf using synchrotron radiation. AB - Enhanced decay of the 31-yr isomer of (178)Hf induced by x-ray irradiation has been reported previously. Here we describe an attempt to reproduce this result with an intense "white" x-ray beam from the Advanced Photon Source. No induced decay was observed. The upper limits for the energy-integrated cross sections for such a process, over the range of energies of 20--60 keV x rays, are less than 2 x 10(-27) cm(2) keV, below the previously reported values by more than 5 orders of magnitude; at 8 keV the limit is 5 x 10(-26) cm(2) keV. PMID- 11497888 TI - Strong enhancement of extremely energetic proton production in central heavy ion collisions at intermediate energy. AB - The energetic proton emission has been investigated as a function of the reaction centrality for the system (58)Ni + (58)Ni at 30A MeV. Extremely energetic protons (E(NN)(p) > or = 130 MeV) were measured and their multiplicity is found to increase almost quadratically with the number of participant nucleons, thus indicating the onset of a mechanism beyond one- and two-body dynamics. PMID- 11497889 TI - Excitation of electronic states of Ar, H(2), and N(2) by positron impact. AB - We have measured the first state-resolved, absolute cross sections for positron excitation of electronic states of an atom or molecule using a high resolution (Delta E approximately 25 meV FWHM) beam of positrons from a Penning-Malmberg trap. We present cross sections for the excitation of the low-lying levels of Ar, H(2), and N(2) for incident positron energies between threshold and 30 eV. For Ar and H2, comparison can be made with theoretical calculations, and, in the case of H(2), the results resolve a significant discrepancy between the only two available calculations. PMID- 11497890 TI - Dissociation energy for C(2) loss from fullerene cations in a storage ring. AB - We have stored positively charged fullerene ions C(+)(n) (n even, from 48 to 70 and 76), C(2+)(60) and C(2+)(70) in an electrostatic storage ring and have measured the rate of emission of neutral fragments as a function of time. In the time range of the measurements, 50 micros to a few milliseconds, the rate decreases strongly due to radiative cooling of the molecules. Using the cooling rate predicted from a dielectric model, we have extracted the dissociation energies for C(2) loss from the measurements. As expected, the energies are largest for the "magic" fullerenes, C(50), C(60), and C(70), and the value of 9.8+/-0.1 eV for C(2) loss from C(+)(60) is in reasonable agreement with theory and with other recent experiments. PMID- 11497891 TI - Enhanced Kerr nonlinearity via atomic coherence in a three-level atomic system. AB - We measure the Kerr-nonlinear index of refraction of a three-level Lambda-type atomic system inside an optical ring cavity. The Kerr nonlinearity is modified and greatly enhanced near atomic resonant conditions for both probe and coupling beams. The Kerr nonlinear coefficient n(2) changes sign when the coupling beam frequency detuning switches sign, which can lead to interesting applications in optical devices such as all-optical switches. PMID- 11497892 TI - Transition to coherence in populations of coupled chaotic oscillators: a linear response approach. AB - We consider the collective dynamics in an ensemble of globally coupled chaotic maps. The transition to the coherent state with a macroscopic mean field is analyzed in the framework of the linear response theory. The linear response function for the chaotic system is obtained using the perturbation approach to the Frobenius-Perron operator. The transition point is defined from this function by virtue of the self-excitation condition for the feedback loop. Analytical results for the coupled Bernoulli maps are confirmed by the numerics. PMID- 11497893 TI - Quantum enhancement of momentum diffusion in the delta-kicked rotor. AB - We present detailed observations of the quantum delta-kicked rotor in the vicinity of a quantum resonance. Our experiment consists of an ensemble of cold cesium atoms subject to a pulsed off-resonant standing wave of light. We measure the mean energy and show clearly that at the quantum resonance it is a local maximum. We also examine the effect of noise on the system and find that the greatest sensitivity to this occurs at the resonances. This makes these regions ideal for examining quantum-classical correspondence. A picture based on diffraction is developed which allows the experiments to be readily understood. PMID- 11497894 TI - Thermal insulating behavior in crystals at high frequencies. AB - When solving heat-conduction problems with periodic temperature perturbations, the thermal conductivity is assumed to remain frequency independent. We, however, show by using the molecular dynamics technique and the fluctuation-dissipation theorem a decrease of the effective thermal conductivity of 2 orders of magnitude when the excitation frequency approaches or exceeds the reverse of the phonon mean relaxation time. Most of the dielectric and semiconductor materials have to be considered as strongly insulating in those conditions. The comparison between molecular-dynamics simulations performed in Si crystals and theoretical predictions reveals a clear agreement. PMID- 11497895 TI - Focusing of noncircular self-similar shock waves. AB - We study the focusing of noncircular shock waves in a perfect gas. We construct an explicit self-similar solution by combining three convergent plane waves with regular shock reflections between them. We then show, with a numerical Riemann solver, that there are initial conditions with smooth shocks whose intermediate asymptotic stage is described by the exact solution. Unlike the focusing of circular shocks, our self-similar shocks have bounded energy density. PMID- 11497896 TI - Unlimited relativistic shock surfing acceleration. AB - Nonrelativistic shock surfing acceleration at quasiperpendicular shocks is usually considered to be a preacceleration mechanism for slow pickup ions to initiate diffusive shock acceleration. In shock surfing, the particle accelerates along the shock front under the action of the convective electric field of the plasma flow. However, the particle also gains kinetic energy normal to the shock and eventually escapes downstream. We consider the case when ions are accelerated to relativistic velocities. In this case, the ions are likely to be trapped for infinitely long times, because the energy of bounce oscillations tends to decrease during acceleration. This suggests the possibility of unlimited acceleration by shock surfing. PMID- 11497897 TI - Experimental evidence of surface-plasmon coupling in anisotropic hollow nanoparticles. AB - We report on the investigation of surface-plasmon excitation of anisotropic WS(2) hollow nanoparticles in a near-field geometry by means of a scanning transmission electron microscope. The shell thickness influence on the electron-energy-loss spectroscopy spectra is experimentally observed and is analyzed within a classical dielectric formalism. As for the isotropic case, we evidence one symmetric (tangential) and one antisymmetric (radial) mode. We point out the intriguing fact that, for the anisotropic case, one can relate these modes to the interband transition of the in-plane component of the dielectric tensor and to the bulk-plasmon energy of the out-of-plane component. PMID- 11497898 TI - Simulating particle dispersions in nematic liquid-crystal solvents. AB - A new method is presented for mesoscopic simulations of particle dispersions in nematic liquid-crystal solvents. It allows efficient first-principles simulations of the dispersions involving many particles with many-body interactions mediated by the solvents. A simple demonstration is shown for the aggregation process of a two dimensional dispersion. PMID- 11497899 TI - Mechanical instability and ideal shear strength of transition metal carbides and nitrides. AB - The ideal shear strength of transition metal carbides and nitrides is calculated with the use of the ab initio pseudopotential density functional method. The microscopic mechanism that limits the ideal strength is studied using full atomic and structural relaxation and the results of electronic structure calculations. It is shown that plasticity in perfect crystals can be triggered by electronic instabilities at finite strains. Our study explicitly demonstrates that the ideal strength in these materials is limited by the elastic instability which is in turn initiated by electronic instabilities. The potential application of alloy hardening due to the onset of instabilities at different strains is also discussed. PMID- 11497900 TI - Crystal structure in nematic emulsion. AB - We describe the experimental observation of a crystal structure formed by glycerol droplets suspended in a nematic liquid crystal. The structure exhibits a high density hexagonal ordering. We have experimentally observed a noticeable interaction between droplets with tangential boundary conditions. Within the scope of known models we discuss the nature of appropriate mechanisms of the interaction. PMID- 11497901 TI - Dislocation kink-pair energetics and pencil glide in body-centered-cubic crystals. AB - When body-centered-cubic crystals undergo plastic deformation, the slip planes are often noncrystallographic. By performing atomistic simulation on the activation pathway of dislocation jumps in bcc iron, we show that the main reason for bcc crystals to exhibit this phenomenon is that one type of kink pair has significantly lower energy than all the other types on the same slip plane. Dislocation motion therefore cannot continue on the same slip plane, and the dislocation has to cross slip onto an intersecting slip plane after each atomic jump. Thus in the long run, the average slip plane would be zigzag and noncrystallographic. PMID- 11497902 TI - New transformations of CO(2) at high pressures and temperatures. AB - CO(2) laser heating of solid CO(2) at pressures between 30 and 80 GPa shows that this compound breaks down to oxygen and diamond along a boundary having a negative P-T slope. This decomposition occurs at temperatures much lower than predicted in theory or inferred from previous experiment. Raman spectroscopy and x-ray diffraction were used as structural probes. At pressures higher than 40 GPa the decomposition is preceded by the formation of a new CO(2) phase (CO(2)-VI). These findings limit the stability of nonmolecular CO(2) phases to moderate temperatures and provide a new topology of the CO(2) phase diagram. PMID- 11497903 TI - Self-organized dynamics on a curved growth interface. AB - We experimentally address long-time dynamics of an artificially curved growth interface in directional solidification. Repetitive cell nucleations are found to appear in a disordered way, but to eventually organize themselves in a coherent way, for long times. This behavior is recovered by simulation of a nonlinear advection-diffusion model for phase dynamics. The existence of a periodic attractor is supported by the derivation of a Lyapunov functional for this model. PMID- 11497904 TI - Geometrical and compositional structure at metal-oxide interfaces: MgO on Fe(001). AB - The geometric structure of MgO deposited on Fe(001) in ultrahigh vacuum by electron evaporation was determined in detail by using surface x-ray diffraction. In contrast to the common belief that MgO grows in direct contact on the Fe(001) substrate, we find an FeO interface layer between the substrate and the growing MgO structure which has not been considered thus far. This result opens new perspectives for the understanding of the Fe/MgO/Fe(001) interface and the tunneling magnetoresistance effect in general. PMID- 11497905 TI - Surface and bulk normal state transport properties in K(3)C(60). AB - The temperature dependence of the surface resistivity for a metallic K(3)C(60) ordered film in the nonsuperconducting state has been obtained by reflection electron energy loss spectroscopy. We demonstrate that the normal state electronic and transport properties of the top molecular layer of K(3)C(60) are similar to the corresponding properties measured with bulk sensitive techniques. These observations strengthen and give a general character to the experimental results obtained with surface sensitive techniques on fullerene compounds. In addition, the transport properties may deviate from the Fermi-liquid behavior above 500 K. PMID- 11497906 TI - Quantum mechanical repulsion of exciton levels in a disordered quantum well. AB - Spatially resolved photoluminescence spectra of a single quantum well are recorded by near-field spectroscopy. A set of over four hundred spectra displaying sharp emission lines from localized excitons is subject to a statistical analysis of the two-energy autocorrelation function. An accurate comparison with a quantum theory of the exciton center-of-mass motion in a two dimensional spatially correlated disordered potential reveals clear signatures of quantum mechanical energy level repulsion, giving the spatial and energetic correlations of excitons in disordered quantum systems. PMID- 11497907 TI - Surface states and negative electron affinity in polyethylene. AB - First-principles calculations are used to investigate the electronic properties of the surfaces of polyethylene. The calculations support the experimental evidence of a negative electron affinity, with calculated values of -0.17 eV and 0.10 eV for surfaces with chains perpendicular and parallel to the surface normal, respectively. Both surfaces exhibit a surface state with binding energy 1.2 +/- 0.5 eV with respect to the bulk polyethylene conduction band minimum. Implications of these findings on spectroscopy, as well as on the transport and aging properties of polyethylene for high-voltage applications, are discussed. PMID- 11497908 TI - Mixed lattice and electronic states in high-temperature superconductors. AB - Inelastic neutron scattering measurements are presented which show the abrupt development of new oxygen lattice vibrations near the doping-induced metal insulator transition in La(2--x)Sr(x)CuO(4). A direct correlation is established between these lattice modes and the electronic susceptibility (as measured by photoemission) inferring that such modes mix strongly with charge fluctuations. This electron-lattice coupling can be characterized as a localized one dimensional response of the lattice to short-ranged metallic charge fluctuations. PMID- 11497909 TI - Ratchet effect: demonstration of a relativistic fluxon diode. AB - We report the observation of the ratchet effect for a relativistic flux quantum trapped in an annular Josephson junction embedded in an inhomogeneous magnetic field. In such a solid state system, mechanical quantities are proportional to electrical quantities, so that the ratchet effect represents the realization of a relativistic-flux-quantum-based diode. Mean static voltage response, equivalent to directed fluxon motion, is experimentally demonstrated in such a diode for deterministic as well as stochastic oscillating current forcing. PMID- 11497910 TI - Superconductivity of ternary silicide with the AlB(2)-type structure Sr(Ga(0.37),Si(0.63))(2). AB - A ternary silicide Sr(Ga(0.37),Si(0.63))(2) was synthesized by a floating zone method. Electron diffraction and powder x-ray diffraction measurements indicate that the silicide has the AlB(2)-type structure with the lattice constants of a = 4.1427(6) A and c = 4.7998(9) A, where Si and Ga atoms are arranged in a chemically disordered honeycomb lattice and Sr atoms are inercalated between them. The silicide is isostructural with the high-temperature superconductor MgB(2) reported recently. Electrical resistivity and dc magnetization measurements revealed that it is a type-II superconductor with onset temperature of 3.5 K. PMID- 11497911 TI - Superfluid density in the d-density-wave scenario. AB - Recently Chakravarty, Laughlin, Morr, and Nayak [Phys. Rev. B 62, 4880 (2000)] made an interesting proposal that the cuprate superconductors possess a hidden " d-density-wave" (DDW) order. We study the implication of this proposal for the superfluid density rho(s). We find that it predicts a temperature gradient [d rho(s)/dT](T = 0) that is strongly doping dependent near the critical doping at which the superconducting gap vanishes. This demonstrates that the DDW scenario is inconsistent with existing well-established experimental data. PMID- 11497912 TI - Interfaces and the question of regional congruence in spin glasses. AB - We present a general theorem restricting properties of interfaces between thermodynamic states and apply it to the spin glass excitations observed numerically by Krzakala and Martin and separately by Palassini and Young in spatial dimensions d = 3,4. We show that such excitations, with interface dimension d(s) < d, cannot yield regionally congruent thermodynamic states. More generally, zero density interfaces of translation-covariant excitations cannot be pinned (by the disorder) in any d but rather must deflect to infinity in the thermodynamic limit. Additional consequences concerning regional congruence in spin glasses and other systems are discussed. PMID- 11497913 TI - Spin-orbit coupling in the Mott insulator Ca(2)RuO(4). AB - O 1s x-ray absorption study of the Mott insulator Ca(2)RuO(4) shows that the orbital population of the 4d t(2g) band dramatically changes with temperature. In addition, spin-resolved circularly polarized photoemission study of Ca(2)RuO(4) shows that a substantial orbital angular momentum is induced in the Ru 4d t(2g) band. Based on the experimental results and model Hartree-Fock calculations, we argue that the cooperation between the strong spin-orbit coupling in the Ru 4d t(2g) band and the small distortion of the RuO(6) octahedra causes the interesting changeover of the spin and orbital anisotropy as a function of temperature. PMID- 11497914 TI - X-ray speckle contrast variation across absorption edges. AB - We measured static x-ray speckle contrast variation with the incident photon energy across a sample-specific absorption edge. In this paper, we present a theoretical description of this energy dependency consistent with our data. We found that the contrast depends mainly on the imaginary part of the complex index of refraction in the sample, as well as on the instrumental resolution. The speckle contrast decreases as the absorption cross section in the sample increases at the absorption edge. This result is not predicted by commonly used theory. PMID- 11497915 TI - Three-dimensional ab initio quantum dynamics of the photodesorption of CO from Cr(2)O(3)(0001): stereodynamic effects. AB - Having performed the first three-dimensional ab initio quantum dynamical study of photodesorption from solid surfaces, we gained mechanistic understanding of the rotational alignment observed in the CO/Cr(2)O(3)(0001) system. Our study is based on potential energy surfaces obtained by embedded cluster calculations for both the electronic ground and excited state of the adsorbate substrate complex. Stochastic wave packet calculations demonstrate the importance of the angular degrees of freedom for the microscopic picture of the desorption process in addition to the desorption coordinate. PMID- 11497916 TI - Radio frequency magnetic field effects on electron-hole recombination. AB - We present measurements of the spectrum (1--80 MHz) of the effect of a weak (approximately 500 microT) radio frequency magnetic field on the electron-hole recombination of radical ion pairs in solution. Distinct spectra are observed for the pyrene anion/dimethylaniline cation radical pair in which one or both of the radicals are perdeuterated. The radical pair mechanism is developed theoretically and shown to account satisfactorily for both the magnetic field effect and the associated magnetic isotope effect. PMID- 11497917 TI - Virtual quantum subsystems. AB - The physical resources available to access and manipulate the degrees of freedom of a quantum system define the set A of operationally relevant observables. The algebraic structure of A selects a preferred tensor product structure, i.e., a partition into subsystems. The notion of compoundness for quantum systems is accordingly relativized. Universal control over virtual subsystems can be achieved by using quantum noncommutative holonomies PMID- 11497918 TI - Remote state preparation. AB - Quantum teleportation uses prior entanglement and forward classical communication to transmit one instance of an unknown quantum state. Remote state preparation (RSP) has the same goal, but the sender knows classically what state is to be transmitted. We show that the asymptotic classical communication cost of RSP is one bit per qubit--half that of teleportation--and even less when transmitting part of a known entangled state. We explore the tradeoff between entanglement and classical communication required for RSP, and discuss RSP capacities of general quantum channels. PMID- 11497919 TI - Requirement of optical coherence for continuous-variable quantum teleportation. AB - We show that the sender and the receiver each require coherent devices in order to achieve unconditional continuous variable quantum teleportation (CVQT), and this requirement cannot be achieved with conventional laser sources, linear optics, ideal photon detectors, and perfect Fock state sources. The appearance of successful CVQT in recent experiments is due to interpreting the measurement record fallaciously in terms of one preferred ensemble (or decomposition) of the correct density matrix describing the state. Our analysis is unrelated to technical problems such as laser phase drift or finite squeezing bandwidth. PMID- 11497920 TI - Multilayer neural networks with extensively many hidden units. AB - The information processing abilities of a multilayer neural network with a number of hidden units scaling as the input dimension are studied using statistical mechanics methods. The mapping from the input layer to the hidden units is performed by general symmetric Boolean functions, whereas the hidden layer is connected to the output by either discrete or continuous couplings. Introducing an overlap in the space of Boolean functions as order parameter, the storage capacity is found to scale with the logarithm of the number of implementable Boolean functions. The generalization behavior is smooth for continuous couplings and shows a discontinuous transition to perfect generalization for discrete ones. PMID- 11497921 TI - Spatiotemporal symmetry in rings of coupled biological oscillators of Physarum plasmodial slime mold. AB - Spatiotemporal patterns in rings of coupled biological oscillators of the plasmodial slime mold, Physarum polycephalum, were investigated by comparing with results analyzed by the symmetric Hopf bifurcation theory based on group theory. In three-, four-, and five-oscillator systems, all types of oscillation modes predicted by the theory were observed including a novel oscillation mode, a half period oscillation, which has not been reported anywhere in practical systems. Our results support the effectiveness of the symmetric Hopf bifurcation theory in practical systems. PMID- 11497922 TI - Bioastrophysical aspects of low energy ion irradiation of frozen anthracene containing water. AB - The origin of life on Earth remains a fascinating mystery in spite of many theories existing on this subject. However, it seems that simple prebiotic molecules could play an essential role in the formation of more complex organisms. In our experiment, we synthesized a class of these molecules (quinones) bombarding frozen anthracene containing water with low energy hydrogen ions. This experiment roughly simulated the astrophysical conditions which one can find in the solar system. Thus, we can hypothesize that prebiotic molecules could be created by interaction of the solar wind with interplanetary dust grains. The delivery of these molecules to early Earth may have contributed to the generation of life on our planet. PMID- 11497923 TI - Free energy self-averaging in protein-sized random heteropolymers. AB - Current theories of heteropolymers are inherently macroscopic, but are applied to mesoscopic proteins. To compute the free energy over sequences, one assumes self averaging--a property established only in the macroscopic limit. By enumerating the states and energies of compact 18, 27, and 36mers on a lattice with an ensemble of random sequences, we test the self-averaging approximation. We find that fluctuations in the free energy between sequences are weak, and that self averaging is valid at the scale of real proteins. The results validate sequence design methods which exponentially speed up computational design and simplify experimental realizations. PMID- 11497924 TI - Histone core slips along DNA and prefers positioning at the chain end. AB - We studied the stability and dynamics of a model of a nucleosome, the fundamental unit for the packing of long DNA in eukaryotes, using a Brownian dynamics simulation. For the proper folding of a stiff polymer on a core particle, moderate attractive interaction is shown to be essentially important, which explains the empirical experimental protocol for the reconstitution of nucleosomes. The effect of the chain end on the positioning of the core particle is examined and compared with the experimental data by atomic force microscopy measurement. It is also suggested that the core particle exhibits sliding motion along the chain as a manifestation of Brownian motion. PMID- 11497925 TI - Binding of similarly charged plates with counterions only. AB - Similarly and highly charged plates in the presence of multivalent counterions attract each other and form electrostatically bound states. Using Monte-Carlo simulations, we obtain the interplate pressure in the global parameter space. The equilibrium plate separation, where the pressure changes from attractive to repulsive, exhibits a novel unbinding transition. A systematic and asymptotically exact strong-coupling field theory yields the bound state from a competition between counterion entropy and electrostatic attraction, in agreement with simple scaling arguments and simulations. PMID- 11497926 TI - Regular wave propagation out of noise in chemical active media. AB - A pacemaker, regularly emitting chemical waves, is created out of noise when an excitable photosensitive Belousov-Zhabotinsky medium, strictly unable to autonomously initiate autowaves, is forced with a spatiotemporal patterned random illumination. These experimental observations are also reproduced numerically by using a set of reaction-diffusion equations for an activator-inhibitor model, and further analytically interpreted in terms of genuine coupling effects arising from parametric fluctuations. Within the same framework we also address situations of noise-sustained propagation in subexcitable media. PMID- 11497927 TI - Effects of cosmic rays on atmospheric chlorofluorocarbon dissociation and ozone depletion. AB - Data from satellite, balloon, and ground-station measurements show that ozone loss is strongly correlated with cosmic-ray ionization-rate variations with altitude, latitude, and time. Moreover, our laboratory data indicate that the dissociation induced by cosmic rays for CF(2)Cl(2) and CFCl(3) on ice surfaces in the polar stratosphere at an altitude of approximately 15 km is quite efficient, with estimated rates of 4.3 x 10(-5) and 3.6 x 10(-4) s(-1), respectively. These findings suggest that dissociation of chlorofluorocarbons by capture of electrons produced by cosmic rays and localized in polar stratospheric cloud ice may play a significant role in causing the ozone hole. PMID- 11497928 TI - Spin correlation and discrete symmetry in spinor Bose-Einstein condensates. AB - We study spin correlations in Bose-Einstein condensates of spin 1 bosons with scatterings dominated by a total spin equal 2 channel. We show that the low energy spin dynamics in the system can be mapped into an o(n) nonlinear sigma model. n = 3 at the zero magnetic field limit and n = 2 in the presence of weak magnetic fields. In an ordered phase, the ground state has a discrete Z2 symmetry and is degenerate under the group [U(1)xS(n-1)]/Z(2). We explore consequences of the discrete symmetry and propose some measurements to probe it. PMID- 11497929 TI - Observation of vortex phase singularities in Bose-Einstein condensates. AB - We have observed phase singularities due to vortex excitation in Bose-Einstein condensates. Vortices were created by moving a laser beam through a condensate. They were observed as dislocations in the interference fringes formed by the stirred condensate and a second unperturbed condensate. The velocity dependence for vortex excitation and the time scale for re-establishing a uniform phase across the condensate were determined. PMID- 11497930 TI - Quasipure Bose-Einstein condensate immersed in a Fermi sea. AB - We report the observation of coexisting Bose-Einstein condensate (BEC) and Fermi gas in a magnetic trap. With a very small fraction of thermal atoms, the 7Li condensate is quasipure and in thermal contact with a 6Li Fermi gas. The lowest common temperature is 0.28 microK approximately 0.2(1)T(C) = 0.2(1)T(F) where T(C) is the BEC critical temperature and T(F) the Fermi temperature. The 7Li condensate has a one-dimensional character. PMID- 11497931 TI - Auger test of the Cen A model of highest energy cosmic rays. AB - If, as recently proposed by Farrar and Piran, Cen A is the source of cosmic rays detected above the Greisen-Zatsepin-Kuz'min cutoff, neutrons are approximately 140 more probable than protons to be observed along its line of sight. This is because the proton flux is rendered nearly isotropic by O(microG) intergalactic magnetic fields. With the anticipated aperture of the Southern Auger Observatory, one may expect on the order of 2 neutron events /year above 10(20) eV in the line of sight of Cen A. PMID- 11497932 TI - Position-space description of the cosmic microwave background and its temperature correlation function. AB - We suggest that the cosmic microwave background (CMB) temperature correlation function C(theta) as a function of angle provides a direct connection between experimental data and the fundamental cosmological quantities. The evolution of inhomogeneities in the prerecombination universe is studied using Green's functions in position space. We find that a primordial adiabatic point perturbation propagates as a sharp-edged spherical acoustic wave. Density singularities at its wave fronts create a feature in the CMB correlation function distinguished by a dip at theta approximately 1.2 degrees. Characteristics of the feature are sensitive to the values of cosmological parameters, in particular to the total and the baryon densities. PMID- 11497933 TI - Shear viscosity of strongly coupled N = 4 supersymmetric Yang-Mills plasma. AB - Using the anti-de Sitter/conformal field theory correspondence, we relate the shear viscosity eta of the finite-temperature N = 4 supersymmetric Yang-Mills theory in the large N, strong-coupling regime with the absorption cross section of low-energy gravitons by a near-extremal black three-brane. We show that in the limit of zero frequency this cross section coincides with the area of the horizon. From this result we find eta = pi / 8N(2)T3. We conjecture that for finite 't Hooft coupling g(2)(YM)N the shear viscosity is eta = f(g(2)(YM)N)N2T3, where f(x) is a monotonic function that decreases from O(x(-2)ln(-1)(1/x)) at small x to pi/8 when x-->infinity. PMID- 11497935 TI - Multijet cross sections in deep inelastic scattering at next-to-leading order. AB - We present the perturbative prediction for three-jet production cross section in deep inelastic scattering at the next-to-leading order accuracy. We study the dependence on the renormalization and factorization scales of exclusive three-jet cross section. The perturbative prediction for the three-jet differential distribution as a function of the momentum transfer is compared to the corresponding data obtained by the H1 Collaboration at HERA. PMID- 11497934 TI - Measurement of the electric form factor of the neutron through d-->(e-->,e(')n)p at Q2 = 0.5 (GeV/c)(2). AB - We report the first measurement using a solid polarized target of the neutron electric form factor G(n)(E) via d-->(e-->,e(')n)p. G(n)(E) was determined from the beam-target asymmetry in the scattering of longitudinally polarized electrons from polarized deuterated ammonia ( 15ND3). The measurement was performed in Hall C at Thomas Jefferson National Accelerator Facility in quasifree kinematics with the target polarization perpendicular to the momentum transfer. The electrons were detected in a magnetic spectrometer in coincidence with neutrons in a large solid angle segmented detector. We find G(n)(E) = 0.04632+/-0.00616(stat)+/ 0.00341(syst) at Q2 = 0.495 (GeV/c)(2). PMID- 11497936 TI - Diffractively produced charm final states in 800-GeV/c pp collisions. AB - We report the first observation of diffractively produced open charm in 800-GeV/c pp collisions of the type pp-->pD*X. We measure cross sections of sigma(diff)(D*+) = (0.185+/-0.044+/-0.054) (mu)b and sigma(diff)(D(*-)) = (0.174+/-0.034+/-0.029) (mu)b. Our measurements are based on 4.3x10(9) events recorded by FNAL E690 in the fixed-target run of 1991. We compare our results with previous fixed-target charm experiments. PMID- 11497938 TI - Neutron density distributions deduced from antiprotonic atoms. AB - The differences between neutron and proton density distributions at large nuclear radii in stable nuclei were determined. Two experimental methods were applied: nuclear spectroscopy analysis of the antiproton annihilation residues one mass unit lighter than the target mass and the measurements of strong-interaction effects on antiprotonic x rays. Assuming the validity of two-parameter Fermi neutron and proton distributions at these large radii, the conclusions are that the two experiments are consistent with each other and that for neutron rich nuclei it is mostly the neutron diffuseness which increases and not the half density radius. The obtained neutron and proton rms radii differences are in agreement with previous results. PMID- 11497939 TI - Magic numbers in exotic nuclei and spin-isospin properties of the NN interaction. AB - The magic numbers in exotic nuclei are discussed, and their novel origin is shown to be the spin-isospin dependent part of the nucleon-nucleon interaction in nuclei. The importance and robustness of this mechanism is shown in terms of meson exchange, G-matrix, and QCD theories. In neutron-rich exotic nuclei, magic numbers such as N = 8, 20, etc. can disappear, while N = 6, 16, etc. arise, affecting the structure of the lightest exotic nuclei to nucleosynthesis of heavy elements. PMID- 11497940 TI - Displacing Rydberg electrons: the mono-cycle nature of half-cycle pulses. AB - A THz "half-cycle" pulse is a fast ( <1 ps) unipolar pulse, followed by a slow unipolar pulse of opposite polarity. We found that the interaction of such THz pulses with very high Rydberg states results in a displacement of the electron within the atom, while the ionization is strongly suppressed. In classical terms: the first fast unipolar feature corresponds to a start kick of the Rydberg electron, while the second unipolar feature acts as a stop kick. A semiclassical model is presented which qualitatively reproduces the ionization suppression and redistribution. PMID- 11497941 TI - Nonlinear localized waves in a periodic medium. AB - We analyze the existence and stability of nonlinear localized waves in a periodic medium described by the Kronig-Penney model with a nonlinear defect. We demonstrate the existence of a novel type of stable nonlinear band-gap localized state, and also reveal a generic physical mechanism of the oscillatory wave instabilities associated with the band-gap resonances. PMID- 11497942 TI - Two-photon cavity solitons in active optical media. AB - We show that broad-area cascade lasers with no absorbing intracavity elements support the spontaneous formation of two-dimensional bright localized structures in a dark background. These cavity solitons consist of islands of two-photon emission embedded in a background of single-photon emission. We discuss the mechanisms through which these structures are formed and interact, along with their properties and stability. PMID- 11497943 TI - Inwards propagating waves in a limit cycle medium. AB - The existence of a novel inwards propagating wave motion is demonstrated in a limit-cycle medium both for the FitzHugh-Nagumo and for modified Chernyak Starobin-Cohen reaction-diffusion systems. The waves (pulses) are seen to be moving "backwards," that is, towards the point where the triggering pulse was initiated, instead of the regular propagation away from the origin. The feasibility of the phenomenon and some of its features are analyzed. PMID- 11497944 TI - Exact phase diagram for an asymmetric avalanche process. AB - The Bethe ansatz method and an iterative procedure based on detailed balance are used to obtain exact results for an asymmetric avalanche process on a ring. The average velocity of particle flow, v, is derived as a function of the toppling probabilities and the density of particles, rho. As rho increases, the system shows a transition from intermittent to continuous flow, and v diverges at a critical point rho(c) with exponent alpha. The exact phase diagram of the transition is obtained and alpha is found to depend on the toppling rules. PMID- 11497937 TI - Pion Interferometry of square root of (s(NN)) =130 GeV Au + Au collisions at RHIC. AB - Two-pion correlation functions in Au+Au collisions at square root of [s(NN)] = 130 GeV have been measured by the STAR (solenoidal tracker at RHIC) detector. The source size extracted by fitting the correlations grows with event multiplicity and decreases with transverse momentum. Anomalously large sizes or emission durations, which have been suggested as signals of quark-gluon plasma formation and rehadronization, are not observed. The Hanbury Brown-Twiss parameters display a weak energy dependence over a broad range in square root of [s(NN)]. PMID- 11497945 TI - Particle segregation in vibrofluidized beds due to buoyant forces. AB - We utilize two-dimensional discrete element computer simulations to investigate the equilibrium position of an impurity in a vibrofluidized bed of otherwise homogeneous particles. The steady state equilibrium height of the impurity increases with increasing vibration velocity amplitude and decreases with the impurity to the surrounding particle density ratio. A simple model whereby the impurity weight is balanced by a "buoyant" force due to the surrounding particle impacts makes a good prediction of the impurity position. PMID- 11497946 TI - Transition from symmetric to asymmetric scaling function before drop pinch-off. AB - The drop pinch-off at a nozzle is studied experimentally for a glycerin-water mixture in surrounding air. The neck diameter of the fluid shrinks with constant velocity. After a distinct transition point, the shrink velocity switches to a smaller value. Before that transition point, the shape of the neck can well be described by a symmetric scaling function, as obtained from Stokes-flow theory of drop formation. This function gives way to an asymmetric scaling function in the final stage before pinch-off. PMID- 11497947 TI - Second harmonic FEL oscillation. AB - We have produced and measured for the first time second harmonic oscillation in the infrared region by a free electron laser. Although such lasing is ideally forbidden, since the gain of a plane wave is zero on axis for an electron beam perfectly aligned with a wiggler, a transverse mode antisymmetry allows sufficient gain in this experiment for lasing to occur. We lased at pulse rates up to 74.85 MHz and could produce over 4.5 W average and 40 kW peak of IR power in a 40 nm FWHM bandwidth at 2925 nm. In agreement with predictions, the source preferentially lased in a TEM01 mode. PMID- 11497948 TI - Quasisteady high-confinement reversed shear plasma with large bootstrap current fraction under full noninductive current drive condition in JT-60U. AB - A quasisteady reversed shear plasma with a large bootstrap current fraction ( approximately 80%) has been obtained for the first time in the JT-60U tokamak. The shrinkage of reversed shear region was suppressed by the bootstrap current peaked at the internal transport barrier (ITB) layer and the ITBs at a large radius were sustained, which, by combination with an H-mode edge pedestal, resulted in a high confinement or 2.2 times the H-mode scaling for 6 times energy confinement time or 2.7 s. Furthermore, a full noninductive current drive was obtained by the bootstrap current and the beam driven current. PMID- 11497949 TI - Reaching high poloidal beta at Greenwald density with internal transport barrier close to full noninductive current drive. AB - In the ASDEX Upgrade tokamak, high poloidal beta up to beta(pol) = 3 at the Greenwald density with H-mode confinement has been reached. Because of the high beta, the plasma current is driven almost fully noninductively, consisting of 51% bootstrap and 43% neutral beam driven current. To reach these conditions the discharge is operated at low plasma current ( I(P) = 400 kA) and high neutral beam heating power ( P(NBI) = 10 MW). The discharge combines an edge (H mode) and internal transport barrier at high densities without confinement-limiting MHD activities. The extrapolation to higher plasma currents may offer a promising way for an advanced scenario based fusion reactor. PMID- 11497950 TI - Adiabatic electron thermal pressure fluctuations in tokamak plasmas. AB - Electron thermal pressure fluctuations measured in the edge plasma of the Texas Experimental Tokamak Upgrade are a fundamental component of plasma turbulence on both sides of the velocity shear layer. The ratio of specific heats, estimated from fluctuations in electron temperature and electron number density measured simultaneously at the same electrode, indicates that observed fluctuations are adiabatic. The observations are made by means of a novel Langmuir probe technique, the time domain triple-probe method, which concurrently measures multiple plasma properties at each of two electrodes with the temporal and the spatial resolution required to estimate thermodynamic properties in a turbulent plasma. PMID- 11497951 TI - Investigation of ultrafast laser-driven radiative blast waves. AB - We have examined the evolution of cylindrically symmetric blast waves produced by the deposition of femtosecond laser pulses in gas jets. In high- Z gases radiative effects become important. We observe the production of an ionization precursor ahead of the shock front and deceleration parameters below the adiabatic value of 1/2 (for a cylinder), an effect expected when the blast wave loses energy by radiative cooling. Despite significant radiative cooling, the blast waves do not appear to develop thin shell instabilities expected for strongly radiative waves. This is believed to be due to the stabilizing effect of a relatively thick blast wave shell resulting in part from electron thermal conduction effects. PMID- 11497952 TI - Asymmetric high-energy ion emission from argon clusters in intense laser fields. AB - Clusters of 2x10(3) to 4x10(4) Ar atoms are Coulomb exploded in intense (up to 8x10(15) W cm(-2)) laser fields. The dependence of multiply charged argon ion energies on the polarization state of light is probed. A directional asymmetry in the ion-explosion energies is observed for the highest charge states. The ion energy distribution consists of a low-energy isotropic component, and a high energy anisotropic one. The results are discussed in terms of an asymmetric Coulomb-explosion scenario. PMID- 11497953 TI - On the validity of the independent hot-spot model. AB - The results of the independent hot-spot (IHS) model are compared to those of the underlying stochastic amplifier in the regime where the coupling of the amplifier is close to its critical value. The considered case is that of a 1D linear amplifier with at most one hot spot per interaction length. It is shown that the validity of the critical coupling given by the IHS model depends on the correlation function of the pump field and should be discussed in each particular case. The discrepancy between the IHS model and the underlying amplifier is shown to be due to the random fluctuations of the hot-spot field around its dominant, deterministic, component. PMID- 11497954 TI - Flexoelectricity and piezoelectricity: the reason for the rich variety of phases in antiferroelectric smectic liquid crystals. AB - The free energy of antiferroelectric smectic liquid crystals which takes into account polar order explicitly is presented. Steric, van der Waals, piezoelectric, and flexoelectric interactions to the nearest layers, and dipolar electrostatic interactions to the nearest and to the next-nearest layers, induce indirect tilt interactions with chiral and achiral properties, which extend to the third- and to the fourth-nearest layers. Although the strength of microscopic interactions changes monotonically with decreasing temperature, the effective interlayer interactions change nonmonotonically and give rise to a nonmonotonic change of the modulation period through various phases. Increased chirality changes the phase sequence. PMID- 11497955 TI - Vibrational spectrum of topologically disordered systems. AB - The topological nature of the disorder of glasses and supercooled liquids strongly affects their high-frequency dynamics. In order to understand its main features, we analytically studied a simple topologically disordered model, where the particles oscillate around randomly distributed centers, interacting through a generic pair potential. We present results of a resummation of the perturbative expansion in the inverse particle density for the dynamic structure factor and density of states. This gives accurate results for the range of densities found in real systems. PMID- 11497956 TI - Two-stage rotational disordering of a molecular crystal surface: C60. AB - We propose a two-stage mechanism for the rotational surface disordering phase transition of a molecular crystal, as realized in C60 fullerite. Our study, based on Monte Carlo simulations, uncovers the existence of a new intermediate regime, between a low-temperature ordered (2x2) state, and a high-temperature (1x1) disordered phase. In the intermediate regime there is partial disorder, strongest for a subset of particularly frustrated surface molecules. These concepts and calculations provide a coherent understanding of experimental observations, with possible extension to other molecular crystal surfaces. PMID- 11497957 TI - Observation of a reentrant twist grain boundary phase. AB - We report the occurrence of a reentrant twist grain boundary phase, which we designate as Re- TGB(A). Microscopic observations on a nonsymmetric dimer showed the phase sequence Iso- N*- TGB(A)-Sm- A-Re- TGB(A)- TGB(C*). Here N* and Sm- A stand for the chiral nematic and smectic- A phases, TGB(A) is the twist grain boundary phase with smectic- A blocks, and TGB(C*) that with smectic- C* blocks and exhibiting features of both the smectic- C* and TGB phases. The reentrance of the TGB(A) phase is unambiguously demonstrated using x-ray diffraction, selective reflection, and optical rotation data. PMID- 11497958 TI - Plasticity and an inverse brittle-to-ductile transition in strontium titanate. AB - The use of ceramic materials is often restricted by a transition from ductile behavior to brittle fracture with decreasing temperature. For example, strontium titanate ( SrTiO3) is known to be extremely fragile and brittle below 1300 K. It is therefore surprising to find that SrTiO3 single crystals can be deformed in compression below 1050 K again. Extensive plastic deformation up to 7% strain at low yield stresses of the order of only 120 MPa is possible at room temperature. Low temperature plasticity is carried by the same [110] [110] dislocations as the high temperature deformation along the [001] axis. From this we conclude that these dislocations must exist in two different core configurations. PMID- 11497959 TI - Atomic structure of icosahedral B4C boron carbide from a first principles analysis of NMR spectra. AB - Density functional theory is demonstrated to reproduce the 13C and 11B NMR chemical shifts of icosahedral boron carbides with sufficient accuracy to extract previously unresolved structural information from experimental NMR spectra. B4C can be viewed as an arrangement of 3-atom linear chains and 12-atom icosahedra. According to our results, all the chains have a CBC structure. Most of the icosahedra have a B11C structure with the C atom placed in a polar site, and a few percent have a B (12) structure or a B10C2 structure with the two C atoms placed in two antipodal polar sites. PMID- 11497960 TI - Directed polymers with tilted columnar disorder and Burgers-like turbulence. AB - The minimal energy variations of a directed polymer with tilted columnar disorder in two dimensions are shown numerically to obey multiscaling at short distances which crosses over to global simple scaling at large distances. The scenario is analogous to that of structure functions in bifractal Burgers turbulence. Some scaling properties are predicted from extreme value statistics. The multiscaling disappears for zero tilt. PMID- 11497961 TI - Ionic conduction in glass: new information on the interrelation between the "Jonscher behavior" and the "nearly constant-loss behavior" from broadband conductivity spectra. AB - We analyze broadband ac conductivity spectra of various ion conducting glasses with regard to the composition dependence of the "Jonscher behavior" and of the "nearly constant-loss behavior." In the temperature range of our experiments (173 K< or =T< or =573 K), both types of behavior are closely related, indicating that both arise from ion hopping. However, the "nearly constant-loss behavior" observed at temperatures below 100 K seems to be caused by other dynamic processes. PMID- 11497962 TI - Molecular adsorption bond lengths at metal oxide surfaces: failure of current theoretical methods. AB - New experimental structure determinations for molecular adsorbates on NiO(100) reveal much shorter Ni-C and Ni-N bond lengths for adsorbed CO and NH3 as well as NO (2.07, 1.88, 2.07 A) than previously computed theoretical values, with discrepancies up to 0.79 A, highlighting a major weakness of current theoretical descriptions of oxide-molecule bonding. Comparisons with experimentally determined bond lengths of the same species adsorbed atop Ni on metallic Ni(111) show values on the oxide surface that are consistently larger (0.1-0.3 A) than on the metal, indicating somewhat weaker bonding. PMID- 11497963 TI - Novel interface-mediated metastable oxide phases: vanadium oxides on Pd(111). AB - In the growth process of ultrathin films of vanadium oxides on Pd(111), a sequence of novel oxide phases with layer-dependent structures and oscillating oxidation states has been detected experimentally and understood theoretically. These phases are interface mediated and metastable with respect to further oxide growth. Transformation into the stable oxide configuration occurs beyond a critical thickness, where energetics combined with kinetic limitations determine the oxide multilayer structure. PMID- 11497964 TI - Crystallographic structure of ultrathin Fe films on Cu(100). AB - We report the observation of bcc-like crystal structures in 2-4 monolayer (ML) Fe films grown on fcc Cu(100) using scanning tunneling microscopy. The local bcc structure provides a straightforward explanation for their frequently reported outstanding magnetic properties, i.e., ferromagnetic ordering in all layers with a Curie temperature above 300 K. The nonpseudomorphic structure, which becomes pseudomorphic above 4 ML film thickness, is unexpected in terms of conventional rules of thin film growth and stresses the importance of finite thickness effects in ferromagnetic ultrathin films. PMID- 11497965 TI - Coupling length scales for multiscale atomistics-continuum simulations: atomistically induced stress distributions in Si/Si3N4 nanopixels. AB - A hybrid molecular-dynamics (MD) and finite-element simulation approach is used to study stress distributions in silicon/silicon-nitride nanopixels. The hybrid approach provides atomistic description near the interface and continuum description deep into the substrate, increasing the accessible length scales and greatly reducing the computational cost. The results of the hybrid simulation are in good agreement with full multimillion-atom MD simulations: atomic structures at the lattice-mismatched interface between amorphous silicon nitride and silicon induce inhomogeneous stress patterns in the substrate that cannot be reproduced by a continuum approach alone. PMID- 11497966 TI - Scaling of the magnetoconductivity of silicon MOSFETs: evidence for a quantum phase transition in two dimensions. AB - For a broad range of electron densities n and temperatures T, the in-plane magnetoconductivity of the two-dimensional system of electrons in silicon MOSFETs can be scaled onto a universal curve with a single parameter H(sigma)(n,T), where H(sigma) obeys the empirical relation H(sigma) = A(n) [Delta(n)(2)+T2](1/2). The characteristic energy k(B)Delta associated with the magnetic field dependence of the conductivity decreases with decreasing density, and extrapolates to 0 at a critical density n(0), signaling the approach to a zero-temperature quantum phase transition. We show that H(sigma) = AT for densities near n(0). PMID- 11497967 TI - "Devil's staircase"-type phase transition in NaV2O5 under high pressure. AB - The "devil's staircase"-type phase transition in the quarter-filled spin-ladder compound NaV2O5 has been discovered at low temperature and high pressure by synchrotron radiation x-ray diffraction. A large number of transitions are found to successively take place among higher-order commensurate phases with 2a x 2b x zc type superstructures. The observed temperature and pressure dependence of modulation wave number q(c), defined by 1/z, is well reproduced by the axial next nearest neighbor Ising model. The q(c) is suggested to reflect atomic displacements presumably coupled with charge ordering in this system. PMID- 11497968 TI - Voltage-modulated millimeter-wave spectroscopy on a polymer diode: mesoscopic charge transport in conjugated polymers. AB - We present a new technique to determine the carrier mobility mu in semiconducting, undoped, conjugated polymers in the millimeter-wave frequency range, 10-500 GHz. This method probes charge conduction on mesoscopic length scales, a regime inaccessible to other transport experiments. The experiment is based on the detection of millimeter-wave absorption of field-induced charge in polymer diodes, and is applied to poly( p-phenylene vinylene). We demonstrate that locally mu can be as high as 10(-4) m(2)/V s, and deduce the typical hopping parameters in the mesoscopic high- mu regions in the material. PMID- 11497969 TI - Indication of the ferromagnetic instability in a dilute two-dimensional electron system. AB - The magnetic field B(c), in which the electrons become fully spin polarized, is found to be proportional to the deviation of the electron density from the zero field metal-insulator transition in a two-dimensional electron system in silicon. The tendency of B(c) to vanish at a finite electron density suggests a ferromagnetic instability in this strongly correlated electron system. PMID- 11497970 TI - Strong friction limit in quantum mechanics: the quantum Smoluchowski equation. AB - For a quantum system coupled to a heat bath environment the strong friction limit is studied starting from the exact path integral formulation. Generalizing the classical Smoluchowski limit to low temperatures, a time evolution equation for the position distribution is derived and the strong role of quantum fluctuations in this limit is revealed. PMID- 11497971 TI - Disorder and quantum fluctuations in superconducting films in strong magnetic fields. AB - We find that the upper critical field in a two-dimensional disordered superconductor can increase essentially at low temperatures. This happens due to the formation of local superconducting islands weakly coupled via the Josephson effect. The distribution of the superconducting islands is derived. It is shown that the value of the critical field is determined by the interplay of the proximity effect and quantum phase fluctuations. The shift of the upper critical field is connected with the pinning properties of a superconductor. PMID- 11497972 TI - High current-carrying capability in c-axis-oriented superconducting MgB2 thin films. AB - In high-quality c-axis-oriented MgB2 thin films, we observed high critical current densities ( J(c)) of approximately 16 MA/cm(2) at 15 K under self-fields comparable to those of cuprate high-temperature superconductors. The extrapolated value of J(c) at 5 K was estimated to be approximately 40 MA/cm(2). For a magnetic field of 5 T, a J(c) of approximately 0.1 MA/cm(2) was detected at 15 K, suggesting that this compound would be a very promising candidate for practical applications at high temperature and lower power consumption. The vortex-glass phase is considered to be a possible explanation for the observed high current carrying capability. PMID- 11497973 TI - Evidence of doping-dependent pairing symmetry in cuprate superconductors. AB - Scanning tunneling spectroscopy studies reveal long-range spatial homogeneity and predominantly d(x(2)-y(2))-pairing spectral characteristics in under- and optimally doped YBa2Cu 3O (7-delta) superconductors, whereas STS on YBa2(Cu 0.9934Zn 0.0026Mg (0.004))3O (6.9) exhibits microscopic spatial modulations and strong scattering near the Zn or Mg impurity sites, together with global suppression of the pairing potential. In contrast, in overdoped (Y 0.7Ca (0.3))Ba 2Cu 3O (7-delta), (d(x(2)-y(2))+s)-pairing symmetry is found, suggesting significant changes in the superconducting ground state at a critical doping value. PMID- 11497974 TI - Role of boron p-electrons and holes in superconducting MgB2, and other diborides: a fully relaxed, full-potential electronic structure study. AB - We present the results of fully relaxed, full-potential electronic structure calculations for the new superconductor MgB (2), and BeB (2), NaB (2), and AlB (2), using density-functional-based methods. Our results, described in terms of (i) density of states (DOS), (ii) band structure, and (iii) the DOS and the charge density around the Fermi energy E(F), clearly show the importance of B p band for superconductivity. In particular, we show that around E(F), the charge density in MgB (2), BeB (2), and NaB (2) is planar and is associated with the B plane. For BeB (2) and NaB (2), we find significant differences in their electronic structure due to differences in the number of valence electrons and the lattice constants a and c. PMID- 11497975 TI - Beyond Eliashberg superconductivity in MgB2: anharmonicity, two-phonon scattering, and multiple gaps. AB - Density-functional calculations of the phonon spectrum and electron-phonon coupling in MgB (2) are presented. The E(2g) phonons, which involve in-plane B displacements, couple strongly to the p(x,y) electronic bands. The isotropic electron-phonon coupling constant is calculated to be about 0.8. Allowing for different order parameters in different bands, the superconducting lambda in the clean limit is calculated to be significantly larger. The E(2g) phonons are strongly anharmonic, and the nonlinear contribution to the coupling between the E(2g) modes and the p(x,y) bands is significant. PMID- 11497976 TI - Phase separation in mixtures of hard core bosons. AB - A theoretical investigation of boson versions of the t-J and t-J(z) models on the square lattice is carried out. In the t-J(z) model, phase separation between a hole-rich and a hole-free phase occurs, at sufficiently low hole doping, for arbitrarily small values of J(z). The boson t-J model, instead, features a uniform ground state at any doping for J/t< or =1.5. No evidence of a striped ground state is found. Relevance of this study to the corresponding fermion models is discussed. Fermi statistics is found to enhance the tendency toward phase separation; in particular, phase separation is predicted, at low doping, in the fermion t-J(z) model, at all values of J(z). PMID- 11497977 TI - Perpendicular exchange bias of Co/Pt multilayers. AB - Exchange bias measurements of ferromagnetic/antiferromagnetic (F/AF) bilayers are typically performed with the magnetization of the F layer parallel to the AF interface. We describe measurements of Co/Pt multilayers with out-of-plane magnetic easy axis that are exchange biased with CoO. Field-cooling experiments with the applied field perpendicular and parallel to the sample plane exhibit loop shifts and enhanced coercivities. Modeling and comparison to biasing of samples with planar easy axis suggests such measurements provide a way to probe the spin projections at F/AF interfaces. PMID- 11497978 TI - Spatially inhomogeneous development of antiferromagnetism in URu2Si2: evidence from 29Si NMR under pressure. AB - From 29Si NMR study, we present evidence for spatially inhomogeneous development of antiferromagnetic (AF) ordering below T(o) = 17.5 K in URu2Si2. In the pressure range between 3.0 and 8.3 kbar, we have observed the 29Si NMR lines arising from the AF region as well as the previously observed 29Si NMR line which correspond to the nonmagnetic region in the sample. The AF volume fraction is enhanced by applied pressure, whereas the magnitude of internal field at the Si site remains constant (910 Oe) up to 8.3 kbar. In the AF region, the ordered moment is about an order of magnitude larger than 0.03 mu(B)/U. PMID- 11497979 TI - Real-space application of the mean-field description of spin-glass dynamics. AB - The out of equilibrium dynamics of finite dimensional spin glasses is considered from a point of view going beyond the standard "mean-field theory" versus "droplet picture" debate of the past decades. The main predictions of both theories concerning the spin-glass dynamics are discussed. It is shown, in particular, that predictions originating from mean-field ideas concerning the violations of the fluctuation-dissipation theorem apply quantitatively, provided one properly takes into account the role of a spin-glass coherence length, which plays a central role in the droplet picture. Dynamics in a uniform magnetic field is also briefly discussed. PMID- 11497980 TI - Phase diagram of an asymmetric spin ladder. AB - We investigate an asymmetric zigzag spin ladder with different exchange integrals on both legs using bosonization and renormalization group approaches. When the leg exchange integrals and frustration both are sufficiently small, renormalization group analysis shows that the Heisenberg critical point flows to an intermediate-coupling fixed point with gapless excitations and a vanishing spin velocity. When they are large, a spin gap opens and a dimer liquid is realized. Here, we find a continuous manifold of Hamiltonians with dimer product ground states, interpolating between the Majumdar-Ghosh and sawtooth spin-chain model. PMID- 11497981 TI - Emission of a single conjugated polymer chain isolated in its single crystal monomer matrix. AB - The excitonic luminescence of a highly ordered single conjugated polymer chain is studied by microphotoluminescence. At T< or =10 K it consists of a single Lorentzian line. The linewidth increases linearly with T between 6 and 60 K, from 350 microeV at 6 K, indicating a pure dephasing time of approximately 2 ps. Above 10 K, other neighboring regions along the chain direction start to emit at a slightly higher (by approximately 1 meV) energy. This indicates very small inhomogeneous broadening, very long chains ( > or =10 microm), and a long range and very rapid exciton energy transfer ( >10 microm in <100 ps). PMID- 11497982 TI - Coupling of nonlocal potentials to electromagnetic fields. AB - Nonlocal Hamiltonians are used widely in first-principles quantum calculations; the nonlocality stems from eliminating undesired degrees of freedom, e.g., core electrons. To date, attempts to couple nonlocal systems to external electromagnetic (EM) fields have been heuristic or limited to weak or long wavelength fields. Using Feynman path integrals, we derive an exact, closed-form coupling of arbitrary EM fields to nonlocal systems. Our results justify and clarify the couplings used to date and are essential for systematic computation of linear and especially nonlinear responses. PMID- 11497983 TI - Avalanche dynamics of crackle sound in the lung. AB - We analyze a sequence of short transient sound waves, called "crackles," which are associated with explosive openings of airways during lung inflation. The distribution of time intervals between consecutive crackles Delta(t) shows two regimes of power law behavior. We develop an avalanche model which fits the data over five decades of Delta(t). We find that the regime for large Delta(t) is related to the dynamics of distinct avalanches in a Cayley tree, and the regime for small Delta(t) is determined by the dynamics of crackle propagation within a single avalanche. We also obtain a mean-field solution of the model which provides information about lung inflation. PMID- 11497984 TI - Conformations of proteins in equilibrium. AB - We introduce a simple theoretical approach for an equilibrium study of proteins with known native-state structures. We test our approach with results on well studied globular proteins, chymotrypsin inhibitor (2ci2), barnase, and the alpha spectrin SH3 domain, and present evidence for a hierarchical onset of order on lowering the temperature with significant organization at the local level even at high temperatures. A further application to the folding process of HIV-1 protease shows that the model can be reliably used to identify key folding sites that are responsible for the development of drug resistance. PMID- 11497985 TI - Buckling of actin-coated membranes under application of a local force. AB - The mechanical properties of composite membranes obtained by self-assembly of actin filaments with giant fluid vesicles are studied by micromanipulation with optical tweezers. These complexes exhibit typical mechanical features of a solid shell, including a finite in-plane shear elastic modulus ( approximately 10(-6) N/m). A buckling instability is observed when a localized force of the order of 0.5 pN is applied perpendicular to the membrane plane. Although predicted for polymerized vesicles, this is the first evidence of such an instability. PMID- 11497986 TI - Electrophoretic microrheology in a dilute lamellar phase of a nonionic surfactant. AB - We measured the complex electrophoretic mobility mu(*)(omega) of nanometer-sized particles dispersed in a lyotropic lamellar phase, and observed two relaxation processes corresponding to the two characteristic lengths of lamellar structure. Faster relaxation is caused by the distortion field of lamellar phase induced by the colloidal particles, and slower relaxation is presumably due to the defects in lamellar structure. Since the dynamic transport property is strongly influenced by the microscopic circumstances as shown in this paper, this method is referred to as electrophoretic microrheology. PMID- 11497987 TI - Colloidal interactions in suspensions of rods. AB - We report direct measurements of entropic interactions of colloidal spheres in suspensions of rodlike fd bacteriophage. We investigate the influence of sphere size, rod concentration, and ionic strength on these interactions. Although the results compare favorably with a recent calculation, small discrepancies reveal entropic effects due to rod flexibility. At high salt concentrations, the potential turns repulsive as a result of viral adsorption on the spheres and viral bridging between the spheres. PMID- 11497988 TI - Recording intramolecular mechanics during the manipulation of a large molecule. AB - The technique of single atom manipulation by means of the scanning tunneling microscope (STM) applies to the controlled displacement of large molecules. By a combined experimental and theoretical work, we show that in a constant height mode of manipulation the STM current intensity carries detailed information on the internal mechanics of the molecule when guided by the STM tip. Controlling and time following the intramolecular behavior of a large molecule on a surface is the first step towards the design of molecular tunnel-wired nanorobots. PMID- 11497989 TI - Spatial symmetry breaking in the Belousov-Zhabotinsky reaction with light-induced remote communication. AB - Domains containing spiral waves form on a stationary background in a photosensitive Belousov-Zhabotinsky reaction with light-induced alternating nonlocal feedback. Complex behavior of colliding and splitting wave fragments is found with feedback radii comparable to the spiral wavelength. A linear stability analysis of the uniform stationary states in an Oregonator model reveals a spatial symmetry breaking instability. Numerical simulations show behavior in agreement with that found experimentally and also predict a variety of other new patterns. PMID- 11497990 TI - Interfacial premelting and the thermomolecular force: thermodynamic buoyancy. AB - The presence of a substrate can alter the equilibrium state of another material near their common boundary. Examples include wetting and interfacial premelting. In the latter case, temperature gradients induce spatial variations in the thickness of the premelted film that reflect changes in the strength of the repulsion between the substrate and the solid. We show that the net thermomolecular force on a macroscopic substrate is equivalent to a thermodynamic buoyancy force-proportional to the mass of solid that can occupy the volume enclosed by the substrate and the temperature gradient. PMID- 11497991 TI - Hopping, jumping and looping by restriction enzymes. AB - Type II restriction endonucleases recognize specific DNA sequences and cleave both strands of the DNA at fixed locations at or near their recognition sites. Many of these enzymes are dimeric proteins that recognize, in symmetrical fashion, palindromic DNA sequences. They generally catalyse independent reactions at each recognition site on the DNA, although in some cases they act processively; cutting the DNA first at one site, then translocating along the DNA to another site and cutting that before leaving the DNA. The way in which the degree of processivity varies with the length of DNA between the sites can reveal the mechanism of translocation. In contrast with the common view that proteins move along DNA by 'sliding', the principal mode of transfer of the EcoRV endonuclease is by 'hopping' and 'jumping', i.e. the dissociation of the protein from one site followed by its re-association with another site in the same DNA molecule, either close to or distant from the original site. Other type II restriction enzymes require two copies of their recognition sites for their DNA cleavage reactions. Many of these enzymes, such as SfiI, are tetramers with two DNA-binding surfaces. SfiI has no activity when bound to just one recognition site, and instead both DNA-binding surfaces have to be filled before it becomes active. Although the two sites can be on separate DNA molecules, SfiI acts optimally with two sites on the same DNA, where it traps the DNA between the sites in a loop. SfiI thus constitutes a test system for the analysis of DNA looping. PMID- 11497992 TI - Regulation of clathrin-coated vesicle formation. AB - The formation of clathrin-coated pits at the plasma membrane requires the concerted action of many different molecules. The real challenge lies in determining the hierarchy of these interactions. We are using assays in both intact and permeabilized cells to dissect the temporal requirements for clathrin coated vesicle formation, and also to examine the role of phosphorylation of the coat proteins. PMID- 11497993 TI - Molecular determinants in pleckstrin homology domains that allow specific recognition of phosphoinositides. AB - More than 250 pleckstrin homology (PH) domains have been identified in the human proteome. All PH domains studied to date appear to bind phosphoinositides, most binding only weakly and non-specifically. Members of a small subclass of PH domains show both high affinity and specificity for particular phosphoinositides, and recent structural studies have provided detailed views of these specific interactions. We discuss the architecture of the specific phosphoinositide binding sites of PH domains, and how selectivity can be modulated by sequence changes. PMID- 11497994 TI - The mechanism of protein kinase regulation by protein phosphatases. AB - Protein kinases are an important class of substrate of the protein phosphatases. We have examined the mechanism of dephosphorylation of the activation segments of the insulin receptor kinase and cyclin-dependent kinase 2 by their respective phosphatases, namely the tyrosine specific phosphatase PTP1B and the dual specificity phosphatase KAP. These studies reveal that PTP1B and KAP utilize contrasting mechanisms in order to dephosphorylate their substrates specifically. PMID- 11497995 TI - Basal and regulated transcription in Archaea. AB - The basal transcription machinery of Archaea is fundamentally related to the eucaryal RNA polymerase (RNAP) II apparatus. In addition to a 12-subunit RNAP, Archaea possess two general transcription factors, the activities of which are required for accurate and efficient in vitro transcription. These factors, TBP and TFB, are homologues of the eucaryal TATA-box binding protein and TFIIB respectively. Archaea also possess TFE, a homologue of the eucaryal RNAP II general transcription factor TFIIE. Although not absolutely required for transcription in vitro, TFE nonetheless plays a stimulatory role under conditions where promoter recognition by TBP is sub-optimal. The basal transcription apparatus of Archaea is closely related to that of Eucarya but archaeal transcriptional regulators resemble those of bacteria. The mode of action of two such regulators has been characterized to determine how these 'bacterial-like' regulators impinge on the 'eucaryal-like' basal machinery. PMID- 11497996 TI - HMG1 and 2: architectural DNA-binding proteins. AB - HMG1 and 2 (high mobility group proteins 1 and 2; renamed HMGB1 and 2) contain two DNA-binding HMG-box domains (A and B) and a long acidic C-terminal domain. They bind DNA without sequence specificity, but have a high affinity for bent or distorted DNA, and bend linear DNA. The individual A and B boxes (which, although broadly similar, show both structural and functional differences) exhibit many of the structure-specific properties of the whole protein. The acidic tail modulates the affinity of the tandem HMG boxes in HMG1 and 2 for a variety of DNA targets, including four-way junctions, but not distorted DNA minicircles, to which the proteins bind with very high affinity. HMG1 and 2 appear to play important architectural roles in the assembly of nucleoprotein complexes in a variety of biological processes, for example V(D)J recombination, the initiation of transcription, and DNA repair. PMID- 11497997 TI - Redox signalling in chloroplasts and mitochondria: genomic and biochemical evidence for two-component regulatory systems in bioenergetic organelles. AB - Redox chemistry is central to the primary functions of chloroplasts and mitochondria, that is, to energy conversion in photosynthesis and respiration. However, these bioenergetic organelles always contain very small, specialized genetic systems, relics of their bacterial origin. At huge cost, organellar genomes contain, typically, a mere 0.1% of the genetic information in a eukaryotic cell. There is evidence that chloroplast and mitochondrial genomes encode proteins whose function and biogenesis are particularly tightly governed by electron transfer. We have identified nuclear genes for 'bacterial' histidine sensor kinases and aspartate response regulators that seem to be targeted to chloroplast and mitochondrial membranes. Sequence similarities to cyanobacterial redox signalling components indicate homology and suggest conserved sensory and signalling functions. Two-component redox signalling pathways might be ancient, conserved mechanisms that permit endogenous control over the biogenesis, in situ, of bioenergetic complexes of chloroplasts and mitochondria. PMID- 11497998 TI - Can we identify the forces that drive the folding of integral membrane proteins? AB - Protein folding has been at the forefront of molecular cell biology research for several years. However, integral membrane proteins have eluded detailed molecular level study until recently. One reason is the often apparently insurmountable problem of mimicking the natural membrane bilayer with lipid or detergent mixtures. There is nevertheless a large body of information on lipid properties and in particular on phosphatidylcholine and phosphatidylethanolamine lipids, which are common to many biological membranes. We have exploited this knowledge to design efficient in vitro, lipid-bilayer folding systems for membrane proteins. Bacteriorhodopsin has been used as a model system for our initial studies: we have shown that a rate-limiting apoprotein folding step and the overall folding efficiency seem to be controlled by particular properties of the lipid bilayer. The properties of interest are the stored curvature elastic energy within the bilayer and the lateral pressure that the lipid chains exert on their neighbouring folding protein. These are generic properties of the bilayer that can be achieved with simple mixtures of many types of biological lipid and seem to be important in vivo. PMID- 11497999 TI - Possible role for molecular chaperones in assembly and repair of photosystem II. AB - Genes of the HSP70 chaperone family are induced by light. In Chlamydomonas reinhardtii, the induction of HSP70 (70 kDa heat shock protein) chaperones by light results in a partial protection of photosystem II against damage by photoinhibitory conditions. Underexpression of a chloroplast-localized HSP70 protein caused an increased sensitivity of photosystem II to light. Overexpression of this protein had a protective effect. Fluorescence measurements and studies of the turnover of photosystem II core components suggest that this HSP70 might function in both the protection and the regeneration of photosystem II. This concept is supported by fractionation studies in which the plastid HSP70 was found associated with chloroplast membranes. Because the light-induced elevation of HSP70 levels provides protection for photosystem II, we examined whether the chloroplast is involved in this regulation and found that mutants defective in plastid-localized chlorophyll synthesis, i.e. the insertion of Mg(2+) into protoporphyrin IX are impaired in the induction of HSP70 by light. Exogenous addition of Mg-protoporphyrin in the dark induced the genes. The combined results support a model in which chlorophyll precursors are essential in the signalling from chloroplast to nucleus that regulates the chaperone genes. PMID- 11498000 TI - Incorporation of iron-sulphur clusters in membrane-bound proteins. AB - The completely sequenced genome of the cyanobacterium Synechocystis PCC 6803 contains several open reading frames, of which the deduced amino acid sequences show similarities to proteins known to be involved in FeS cluster synthesis of nitrogenase (Nif proteins) and other FeS proteins (Isc proteins). In this article, the results of our studies on these proteins are summarized and discussed with respect to their relevance in FeS cluster incorporation in chloroplasts. In cyanobacteria, there appears to exist several pathways for FeS cluster synthesis. PMID- 11498001 TI - Assembly-controlled regulation of chloroplast gene translation. AB - Studies of the biogenesis of the photosynthetic protein complexes in the unicellular green alga Chlamydomonas reinhardtii have pointed to the importance of the concerted expression of nuclear and chloroplast genomes. The accumulation of chloroplast- and nuclear-encoded subunits is concerted, most often as a result of the rapid proteolytic disposal of unassembled subunits, but the rate of synthesis of some chloroplast-encoded subunits from photosynthetic protein complexes, designed as CES proteins (Controlled by Epistasy of Synthesis), is regulated by the availability of their assembly partners from the same complex. Cytochrome f, a major subunit of the cytochrome b(6)f complex is a model protein for the study of the CES process. In the absence of subunit IV, another subunit of the cytochrome b(6)f complex, its synthesis is decreased by 90%. This results from a negative autoregulation of cytochrome f translation initiation, mediated by a regulatory motif carried by the C-terminal domain of the unassembled protein [Choquet, Stern, Wostrikoff, Kuras, Girard-Bascou and Wollman (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 4380-4385]. Using site-directed mutagenesis, we have characterized this regulatory motif. We discuss the possible implications regarding the mechanism of the CES process for cytochrome f expression. We have studied the possible generalization of this mechanism to other CES proteins. PMID- 11498002 TI - Degradation of unassembled and damaged thylakoid proteins. AB - To study protein degradation in thylakoid membranes we identified, characterized and cloned thylakoid proteases, and then linked them to known proteolytic processes. Several families of chloroplast proteases were identified and characterized to different extents. FtsH, an ATP-dependent metalloprotease that belongs to the AAA-protein family, was found to be integral to the thylakoid membrane, facing the stroma. It is involved in both the degradation of unassembled subunits of membrane complexes, such as the Rieske Fe-S protein of the cytochrome complex, and the degradation of oxidatively damaged proteins such as the D1 protein of the photosystem II (PS II) reaction centre. Plant genomes contain multiple isomers of this protease but the functional significance of this multiplication is not clear yet. A second protease, the serine ATP-independent DegP, was found to be strongly associated with the luminal side of the thylakoid membrane. Although a specific role has not yet assigned for it, its location suggests that it can degrade luminal soluble proteins as well as luminally exposed regions of thylakoid membrane proteins. PMID- 11498003 TI - AAA proteases of mitochondria: quality control of membrane proteins and regulatory functions during mitochondrial biogenesis. AB - An ubiquitous and conserved proteolytic system regulates the stability of mitochondrial inner membrane proteins. Two AAA proteases with catalytic sites at opposite membrane surfaces form a membrane-integrated quality control system and exert crucial functions during the biogenesis of mitochondria. Their activity is modulated by another membrane-protein complex that is composed of prohibitins. Peptides generated upon proteolysis in the matrix space are transported across the inner membrane by an ATP-binding cassette transporter. The function of these conserved components is discussed in the present review. PMID- 11498004 TI - Structure and function of Pet100p, a molecular chaperone required for the assembly of cytochrome c oxidase in Saccharomyces cerevisiae. AB - The assembly of cytochrome c oxidase in the inner mitochondrial membranes of eukaryotic cells requires the protein products of a large number of nuclear genes. In yeast, some of these act globally and affect the assembly of several respiratory-chain protein complexes, whereas others act in a cytochrome c oxidase specific fashion. Many of these yeast proteins have human counterparts, which when mutated lead to energy-related diseases. One of these proteins, Pet100p, is a novel molecular chaperone that functions to incorporate a subcomplex containing cytochrome c oxidase subunits VII, VIIa and VIII into holo-(cytochrome c oxidase). Here we report the topological disposition of Pet100p in the inner mitochondrial membrane and show that its C-terminal domain is essential for its function as a cytochrome c oxidase-specific 'assembly facilitator'. PMID- 11498005 TI - Mutations inactivating mitochondrial genes in Chlamydomonas reinhardtii. AB - Chlamydomonas reinhardtii is now becoming a useful model for the study of mitochondrial genetics in a photosynthetic organism. The small (15.8 kb) mitochondrial genome C. reinhardtii has been sequenced completely and all the genes have been identified. Several mutants inactivated in mitochondrial genes encoding components of the respiratory complexes I, III and IV have been characterized at the molecular level. Assembly of complex I in several mutant strains and mapping of mitochondrial mutations by recombinational analysis are also described. PMID- 11498006 TI - Assembly of cytochrome c oxidase: what can we learn from patients with cytochrome c oxidase deficiency? AB - Cytochrome c oxidase is an intricate metalloprotein that transfers electrons from cytochrome c to oxygen in the last step of the mitochondrial respiratory chain. It uses the free energy of this reaction to sustain a transmembrane electrochemical gradient of protons. Site-directed mutagenesis studies of bacterial terminal oxidases and the recent availability of refined crystal structures of the enzyme are rapidly expanding the understanding of the coupling mechanism between electron transfer and proton translocation. In contrast, relatively little is known about the assembly pathway of cytochrome c oxidase. Studies in yeast have indicated that assembly is dependent on numerous proteins in addition to the structural subunits and prosthetic groups. Human homologues of a number of these assembly factors have been identified and some are now known to be involved in disease. To dissect the assembly pathway of cytochrome c oxidase, we are characterizing tissues and cell cultures derived from patients with genetically defined cytochrome c oxidase deficiency, using biochemical, biophysical and immunological techniques. These studies have allowed us to identify some of the steps of the assembly process. PMID- 11498007 TI - Chlamydomonas nuclear mutants that fail to assemble respiratory or photosynthetic electron transfer complexes. AB - We are using a molecular-genetic approach to investigate the role of nuclear genes in the biogenesis of the electron transfer complexes of mitochondria and chloroplasts. Our analysis of nuclear mutants of the green alga Chlamydomonas that are defective in respiration or photosynthesis has led to the identification of genes encoding factors required for the expression of specific organellar genes, and genes encoding structural components of the complexes. PMID- 11498008 TI - Auxiliary functions in photosynthesis: the role of the FtsH protease. AB - Oxygenic photosynthesis can be described effectively by using two long-standing models: the Z-scheme and the chemiosmotic hypothesis. However, these models do not reveal the dynamic nature of the thylakoid membrane and the four major complexes that it binds. The composition of the photosynthetic apparatus is continually changing in response to a range of environmental stimuli. In addition, many photosynthetic components have some of the highest turnover rates in Nature. Changes in composition and turnover of photosynthetic components require the degradation of existing and damaged polypeptides and the resynthesis and co-ordinated assembly of new polypeptides and their associated cofactors. This is achieved by several auxiliary functions, including proteolysis, protein targeting and the action of molecular chaperones. Some of the components involved in these functions, such as translocons, chaperones and proteases, have been identified but many of the auxiliary functions of photosynthesis remain uncharacterized. Among the proteases known to be associated with the thylakoids is the zinc metalloprotease FtsH, which might also act as a chaperone. Here we provide an overview of the thylakoid FtsH protease and discuss its role in the maintenance and assembly of the photosynthetic apparatus. PMID- 11498009 TI - Structural evidence for multiple transport mechanisms through the Golgi in the pancreatic beta-cell line, HIT-T15. AB - Accurate data on the three-dimensional architecture of the Golgi is prerequisite for evaluating the mechanisms of transit through this organelle. Here we detail the structure of the Golgi ribbon within part of an insulin-secreting cell in three dimensions at approximately 6 nm resolution. Rapid freezing, freeze substitution and electron tomography were employed. The Golgi in this region is composed of seven cisternae. The cis-most element is structurally intermediate between the endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC) and the cis-most cisterna characterized in three dimensions at high resolution in a normal rat kidney cell [Ladinsky, Mastronarde, McIntosh, Howell and Staehelin (1999) J. Cell Biol. 144, 1135-1149]. There are three trans-cisternae that demonstrate morphological and functional variation. The membrane surface areas and volumes of these elements decrease from cis to trans. The two trans-most cisternae are dissociated from the stack and are fragmented by tubulation. ER closely adheres to and inserts between individual trans-cisternae. Many of the 2119 small, clathrin-negative vesicles that are in close proximity to the Golgi fill the region where trans-cisternae have moved out of register with the ribbon. These data provide evidence that cisternal progression/maturation, trafficking via membrane tubules and vesicle-mediated transport act in concert in the same region of the Golgi ribbon, and suggest an important role for the ER in regulating membrane dynamics at the trans-Golgi. PMID- 11498010 TI - Control of membrane fusion dynamics during regulated exocytosis. AB - The study of regulated exocytosis uniquely allows the direct measurement of intracellular membrane fusion events in real time. We have exploited this to examine factors that regulate not only the extent but also the dynamics of single fusion/release events. The general strategy used has been to assess exocytosis in transiently transfected PC12 or adrenal chromaffin cells. We aimed to design mutant constructs based on in vitro biochemistry, in some cases informed by knowledge of protein structure. Using this approach we have demonstrated an inhibitory role for the putative Rab3 effector Noc2 that requires interaction with Rab3. Using carbon-fibre amperometry on adrenal chromaffin cells, we have demonstrated regulation of the kinetics of single granule release events consistent with changes in fusion pore dynamics and switches between full fusion and 'kiss-and-run' fusion. These studies have demonstrated a late role for cysteine string protein in exocytosis. In addition, they have focused attention on a key role for Munc18 in the regulation of post-fusion events that affect fusion pore dynamics. PMID- 11498011 TI - Function of Hrs in endocytic trafficking and signalling. AB - The hepatocyte growth factor-regulated tyrosine kinase substrate, Hrs, becomes tyrosine-phosphorylated upon the binding of various growth factors and cytokines to their receptors. This protein is essential for ventral folding morphogenesis, and it shares structural similarity with Vps27p, which is involved in vacuolar protein sorting in yeast. Since Hrs is localized to endosomes and has been implicated in the regulation of signal transduction as well as membrane trafficking, it has been regarded as a potential co-ordinator of endosomal receptor sorting and signalling. Here we discuss the possible functions of Hrs in light of its interactions with phosphatidylinositol 3-phosphate and multiple proteins. PMID- 11498012 TI - Relationship between endosomes and lysosomes. AB - Delivery of endocytosed macromolecules to lysosomes occurs by means of direct fusion of late endosomes with lysosomes. This has been formally demonstrated in a cell-free content mixing assay using late endosomes and lysosomes from rat liver. There is evidence from electron microscopy studies that the same process occurs in intact cells. The fusion process results in the formation of hybrid organelles from which lysosomes are re-formed. The discovery of the hybrid organelle has opened up three areas of investigation: (i) the mechanism of direct fusion of late endosomes and lysosomes, (ii) the mechanism of re-formation of lysosomes from the hybrid organelle, and (iii) the function of the hybrid organelle. Fusion has analogies with homotypic vacuole fusion in yeast. It requires syntaxin 7 as part of the functional trans-SNARE [SNAP receptor, where SNAP is soluble N ethylmaleimide-sensitive factor (NSF) attachment protein] complex and the release of lumenal calcium to achieve membrane fusion. Re-formation of lysosomes from the hybrid organelle occurs by a maturation process involving condensation of lumenal content and probably removal of some membrane proteins by vesicular traffic. Lysosomes may thus be regarded as a type of secretory granule, storing acid hydrolases in between fusion events with late endosomes. The hybrid organelle is predicted to function as a 'cell stomach', acting as a major site of hydrolysis of endocytosed macromolecules. PMID- 11498013 TI - Internalization of the epidermal growth factor receptor: role in signalling. AB - The interaction of the activated epidermal growth factor (EGF) receptor (EGFR) with the Src homology 2 (SH2) domain of Grb2 (growth-factor-receptor-bound protein 2) initiates signalling through Ras and mitogen-activated protein kinase. Grb2 can bind EGFR directly or through another SH2-containing protein, Shc. Activation of EGFRs by ligand also triggers rapid endocytosis of EGF-receptor complexes. To analyse the spatial and temporal regulation of EGFR interactions with SH2 domains in living cells, we have combined imaging microscopy with a modified method of measuring fluorescence resonance energy transfer (FRET) on a pixel-by-pixel basis using EGFR fused to cyan fluorescent protein (CFP) in pair with Grb2 or Shc fused to yellow fluorescent protein (YFP). Stimulation by EGF resulted in the recruitment of Grb2-YFP and YFP-Shc to cellular compartments that contained EGFR-CFP, and a large increase in the FRET signal. In particular, FRET measurements indicated that activated EGFR-CFP interacted with YFP-Shc and Grb2 YFP in membrane ruffles and endosomes. These results demonstrate that signalling via EGFRs can occur in the endosomal compartment. Moreover, in contrast with previous biochemical studies, FRET experiments show that a large pool of Grb2 and Shc is associated with EGFRs for a prolonged period after EGF stimulation. PMID- 11498014 TI - Proteins involved in the synaptic organization of AMPA (alpha-amino-3-hydroxy-5 methylisoxazolepropionate) receptors. AB - As well mediating most synaptic transmission in the mammalian central nervous system, alpha-amino-3-hydroxy-5-methylisoxazolepropionate (AMPA) receptors are involved in the processes of synapse formation, stabilization and plasticity. Thus the mechanisms that control the developmental and activity-dependent changes in the functional synaptic expression of AMPA receptors are of fundamental importance. In the last few years dramatic advances have been achieved towards elucidating some of the molecular events involved, and a previously unsuspected complexity of proteins that selectively interact directly or indirectly with individual AMPA receptor subunits have been identified. Here I present an overview of some of the main interacting proteins, and describe what we know about how these may fit into cellular pathways that control AMPA receptor targeting, trafficking and functional expression. PMID- 11498015 TI - The ubiquitin-proteasome pathway regulates lysosomal degradation of the growth hormone receptor and its ligand. AB - The growth hormone (GH) receptor (GHR) is a mammalian plasma membrane protein whose internalization is mediated by the ubiquitin-proteasome pathway. GH internalization and degradation are inhibited when cells are treated with proteasome inhibitors. Here we show that a GHR truncated at residue 369 can enter the cells in the presence of a proteasome inhibitor, but that the subsequent lysosomal degradation of GH is blocked. Lysosomal inhibitors prolong the half life of both receptor and ligand. Experiments with antibodies against different receptor tail sections show that degradation of the GHR cytosolic domain precedes degradation of the extracellular GH-binding domain. A possible role for the ubiquitin-proteasome pathway in the degradation of the receptor and ligand is discussed. PMID- 11498016 TI - Caveolin-1, a putative tumour suppressor gene. AB - Caveolae ('little caves') are plasma membrane specializations of 50-100 nm in diameter, and the caveolins are structural proteins used by cells to form caveolae. We and other investigators have discovered that caveolae organelles may be important both in normal signal transduction and in the pathogenesis of a number of human diseases, such as cancer. Here we describe the functional roles of the caveolin gene family and summarize the evidence that supports a role for caveolae as mediators of a number of cellular signalling processes, including apoptosis. PMID- 11498017 TI - Role of endocytosis in signalling and regulation of G-protein-coupled receptors. AB - Many G-protein-coupled receptors (GPCRs) undergo agonist-induced endocytosis. Endocytosis contributes to distinct processes that regulate the number and functional activity of receptors present in the plasma membrane, contributing to the well described processes of receptor sequestration and down-regulation. Emerging evidence suggests additional functions of endocytosis in mediating GPCR signalling via certain effector pathways, such as mitogen-activated protein kinase modules. The diverse functions of endocytosis raise fundamental questions about the nature of the vesicular carriers and membrane pathways that mediate the endocytic trafficking of specific GPCRs. Insights into the biochemical and functional properties of endocytic vesicles containing internalized opioid and adrenergic receptors will be discussed. Progress towards understanding the mechanisms that control the specificity with which distinct GPCRs are sorted to specialized subpopulations of endocytic vesicles will be highlighted. PMID- 11498018 TI - Multiple pathways for the dynamin-regulated internalization of muscarinic acetylcholine receptors. AB - An important regulatory pathway of G-protein-coupled receptors (GPCRs) is the internalization of receptors into the cell interior. To unravel the molecular mechanisms by which GPCRs are internalized, we have studied the internalization of various members of the family of muscarinic acetylcholine receptors (mAChRs). Using the transient expression system of HEK-293 cells, we showed that the M(1), M(3) and M(4) mAChRs are internalized into clathrin-coated vesicles and recycle back to the plasma membrane. This internalization pathway is dependent on the concerted action of beta-arrestin, c-Src and the GTPase dynamin, which 'catalyses' the budding of clathrin-coated vesicles (and other vesicles) from the plasma membrane. Internalization of the M(2) mAChR (which is highly structurally and functionally related to the M(4) receptor subtype) also requires dynamin, but proceeds in an apparent beta-arrestin-, c-Src- and clathrin-independent manner. Internalized M(2) mAChRs also show virtually no receptor recycling, but are down regulated. This demonstrates that GPCRs can be internalized by multiple dynamin dependent pathways in a highly regulated manner. PMID- 11498019 TI - Signalling and non-caveolar rafts. AB - Rafts are small membrane domains containing discrete subsets of lipids and proteins. Although microscopic raft structures termed 'caveolae' were described nearly 50 years ago, the importance of rafts, particularly signalling within rafts, is only beginning to be understood. Our studies focus on receptor dependent phosphoinositide signalling. Using their characteristic buoyancy in density gradients, we and others found that the epidermal growth factor (EGF) receptor, phosphatidylinositol 4-kinase and phosphoinositides are localized within a caveolin-rich fraction of A431 carcinoma cells. We subsequently found that membrane fragments containing the EGF receptor and most cellular phosphoinositides can be separated from caveolae. Consequently, components of EGF dependent phosphoinositide signalling localize to one or more novel types of raft, the composition of which we are currently determining. A key component is the type II phosphatidylinositol 4-kinase, which, for many years, has proven difficult to purify and clone. We describe our recent purification from rafts and cloning of this elusive enzyme, and discuss how the structure sheds light on the rafting of this enzyme. PMID- 11498020 TI - Specificity in ligand binding and intracellular signalling by insulin and insulin like growth factor receptors. AB - The physiological roles of insulin and insulin-like growth factors (IGFs) are distinct, with insulin acting to regulate cellular uptake and metabolism of fuels, whereas IGFs promote cell growth, survival and differentiation. The only components of signalling pathways known to be unique to insulin and IGFs are their respective receptors, and even these display substantial structural and functional similarity. Specificity of action in vivo must in part reflect relative levels of receptor expression in different tissues. The extent to which the receptors differ in intrinsic signalling capacity remains unclear, but specificity might in principle arise from differences in ligand-binding mechanism or properties of intracellular domains. To identify ligand binding determinants we expressed receptor fragments as soluble proteins. Both N-terminal domains and a C-terminal peptide sequence from the alpha-subunit are essential for ligand binding with moderate affinity. However, binding of ligand with high affinity and specificity requires higher-order structure. To compare signalling capacities, we constructed chimaeras containing intracellular domains of insulin or IGF receptors fused to the extracellular portion of TrkC. Expression and activation of these chimaeras in cell lines reveals subtle differences in signalling and end point responses, which may depend on cell background. PMID- 11498021 TI - Insulin receptor substrate proteins and neuroendocrine function. AB - A family of insulin receptor substrate (IRS) proteins mediates the pleiotropic effects of insulin and insulin-like growth factor 1 (IGF-1) on cellular function by recruiting several intracellular signalling networks. Conventional murine knockout strategies have started to reveal distinct physiological roles for the IRS proteins. Deletion of Irs1 produces a mild metabolic phenotype with compensated insulin resistance but also causes marked growth retardation. In contrast, mice lacking IRS-2 display nearly normal growth but develop diabetes owing to a combination of peripheral insulin resistance and beta-cell failure. As well as the classical metabolic events regulated by insulin signalling pathways, studies in lower organisms have implicated insulin/IGF-1 signalling pathways in the control of food intake and reproductive function. Our analysis of IRS-2 knock out mice shows that female mice are infertile owing to defects in the hypothalamus, pituitary and gonad. IRS-2(-/-) mice have small, anovulatory ovaries with reduced numbers of follicles. Levels of the pituitary hormones luteinizing hormone and prolactin and gonadal steroids are low in these animals. Pituitaries of IRS-2(-/-) animals are decreased in size and contain reduced numbers of gonadotrophs. Additionally, IRS-2(-/-) females display increased food intake and develop obesity, despite elevated leptin levels, suggesting abnormalities in hypothalamic function. Coupled with recent observations that brain-specific deletion of the insulin receptor causes a similar phenotype, these findings implicate IRS signalling pathways in the neuroendocrine regulation of reproduction and energy homeostasis. PMID- 11498022 TI - Functional effects of APS and SH2-B on insulin receptor signalling. AB - APS [for 'adapter protein with a pleckstrin homology (PH) and Src homology 2 (SH2) domain'] belongs to a family of adapter proteins involved in signalling by the receptors for insulin, insulin-like growth factor 1, platelet-derived growth factor and nerve growth factor. Other members include alternatively spliced SH2-B isoforms (SH2Balpha, SH2-Bbeta and SH2-Bgamma) and Lnk. These have a C-terminal SH2 domain, a central PH domain and an N-terminal proline-rich region. SH2Balpha, APS and Lnk have a conserved C-terminal tyrosine phosphorylation site, whereas the alternatively spliced SH2-Bbeta and SH2-Bgamma have distinct C-termini. There is considerable sequence similarity between APS, SH2-B and Lnk, particularly in the SH2 domain. Both APS and SH2-Balpha interact with the insulin-receptor activation loop phosphorylation sites and undergo insulin-stimulated tyrosine phosphorylation, although the phosphorylation of SH2-B is considerably weaker. APS couples c-Cbl to the insulin receptor, resulting in ubiquitination of the insulin receptor. We established cell lines [Chinese hamster ovary (CHO). T-APS and CHO. T-SH2-B cells] overexpressing APS and SH2-Balpha to study their roles in insulin receptor signalling. Either adapter protein enhances insulin receptor and ERK (extracellular-signal-regulated kinase) phosphorylation. In CHO. T-APS cells, Akt phosphorylation is observed earlier than in CHO.T-SH2-B cells. Both enhance insulin-stimulated Akt activation but APS seems to cause greater activation. Thus APS and SH2-B have similar effects on insulin receptor signalling, although the effects of SH2-B are independent of its phosphorylation. PMID- 11498023 TI - Growth factor regulation of the novel class II phosphoinositide 3-kinases. AB - It is well established that the class-I phosphoinositide (PI) 3-kinases play a crucial role in growth factor signalling pathways. However, evidence has recently emerged that the alpha isoform of the class-II PI 3-kinase (PI 3K-C2alpha) is activated by growth factors, although the consequences of this are poorly understood. Here we demonstrate that the activation of PI 3K-C2alpha is not associated with a change in subcellular localization. Furthermore, we provide the first evidence that PI 3K-C2beta is activated by insulin, albeit with slower kinetics than activation of PI 3K-C2alpha. These findings suggest that both these class-II PI 3-kinase isoforms are likely to participate in insulin-signalling pathways in the cell. PMID- 11498024 TI - Regulation of glycogen synthesis in human muscle cells. AB - Glucose uptake into muscle and its subsequent storage as glycogen is a crucial factor in energy homeostasis in skeletal muscle. This process is stimulated acutely by insulin and is impaired in both insulin-resistant states and in type 2 diabetes mellitus. A signalling pathway involving protein kinase B and glycogen synthase kinase 3 seems certain to have a key role in stimulating glycogen synthesis but other signalling pathways also contribute, including a rapamycin sensitive pathway stimulated by amino acids. Although glycogen synthesis is one of the classical insulin-regulated pathways, it is also regulated in an insulin independent manner; for example glycogen synthesis in muscle is stimulated significantly after strenuous exercise, with much of this stimulation being independent of the involvement of insulin. Evidence suggests that glucose and the glycogen content of the muscle have a key role in this stimulation but the molecular mechanism has yet to be fully explained. PMID- 11498025 TI - Interplay between insulin and nutrients in the regulation of translation factors. AB - Protein synthesis in mammalian cells is regulated through alterations in the states of phosphorylation of eukaryotic initiation factors and elongation factors (eIFs and eEFs respectively) and of other regulatory proteins. This modulates their activities or their abilities to interact with one another. Insulin activates several of these proteins including the following: the guanine nucleotide exchange factor eIF2B; the eIF4F complex, which (through eIF4E) interacts with the cap of the mRNA; p70 S6 kinase; and elongation factor eEF2, which mediates the translocation step of elongation. Control of the last three of these is linked to mTOR (mammalian target of rapamycin). In Chinese hamster ovary cells, regulation of all these proteins by insulin is modulated by the presence of amino acids and/or glucose in the medium. For example, p70 S6 kinase activity declines in the absence of amino acids and cannot be stimulated by insulin under this condition. The readdition of amino acids, especially leucine, restores activity and sensitivity to insulin. With eIF2B and eEF2, both amino acids and glucose must be provided for insulin to regulate their activities. In contrast, insulin-stimulation of the formation of eIF4F complexes requires glucose but not amino acids. Glucose metabolism is required for this permissive effect. Our recent studies have also identified the mechanism by which mTOR signalling regulates the phosphorylation of eEF2. eEF2 kinase is phosphorylated by p70 S6 kinase at Ser-366; this results in the inactivation of eEF2 kinase, especially at low (micromolar) Ca concentrations. PMID- 11498026 TI - Sterol-regulatory-element-binding protein 1c mediates insulin action on hepatic gene expression. AB - Effects of insulin on the expression of liver-specific genes are part of the adaptive mechanisms aimed at maintaining energy homeostasis in mammals. When the diet is rich in carbohydrates, secreted insulin stimulates the expression of genes for enzymes involved in glucose utilization (glucokinase, L-type pyruvate kinase and lipogenic enzymes) and inhibits genes for enzymes involved in glucose production (phosphenolpyruvate carboxykinase). The mechanisms by which insulin controls the expression of these genes have been poorly understood. Recently, the transcription factor sterol-regulatory-element-binding protein 1c has been proposed as a key mediator of insulin transcriptional effects. Here we review the evidence that has led to this proposal and the consequences for our understanding of insulin effects in physiological or pathological conditions. PMID- 11498027 TI - Insulin-regulated gene expression. AB - Insulin regulates the expression of more than 150 genes, indicating that this is a major action of this hormone. At least eight distinct consensus insulin response sequence (IRSs) have been defined through which insulin can regulate gene transcription. These include the serum response element, the activator protein 1 ('AP-1') motif, the Ets motif, the E-box motif and the thyroid transcription factor 2 ('TTF-2') motif. All of these IRSs mediate stimulatory effects of insulin on gene transcription. In contrast, an element with the consensus sequence T(G/A)TTT(T/G)(G/T), which we refer to as the phosphoenolpyruvate carboxykinase (PEPCK)-like motif, mediates the inhibitory effect of insulin on transcription of the genes encoding PEPCK, insulin-like growth-factor-binding protein 1 (IGFBP-1), tyrosine aminotransferase and the glucose-6-phosphatase (G6Pase) catalytic subunit. The forkhead transcription factor FKHR has recently been shown to bind this PEPCK-like IRS motif and a model has been proposed in which insulin inhibits gene transcription by stimulating the phosphorylation and nuclear export of FKHR. Our results suggest that this model is consistent with the action of insulin on transcription of the gene encoding IGFBP-1 but not that of the G6Pase catalytic subunit. Thus, even though the IRSs in both promoters seem identical, they are functionally distinct. In addition, in the G6Pase catalytic subunit promoter, hepatocyte nuclear factor 1 ('HNF-1'), acts as an accessory factor to enhance the effect of insulin mediated through the IRS. PMID- 11498028 TI - De novo design, synthesis and characterization of membrane-active peptides. AB - Our current level of understanding of membrane-protein folding is primitive, but it is beginning to advance. Previously [Choma, Gratkowski, Lear and DeGrado (2000) Nat. Struct. Biol. 7, 161-166], we described studies of the association in detergent micelles of short, simple-sequence hydrophobic peptides modified from the sequence of the water-soluble, homodimeric coiled-coil GCN4-P1 peptide using the principle that the interiors of membrane proteins are similar to those of water-soluble proteins. Here, we discuss more quantitative aspects of the association equilibrium and compare the free energies of association of a number of mutant peptides designed to explore specific features responsible for the association. PMID- 11498029 TI - Peptaibols: models for ion channels. AB - Peptaibols are membrane-active polypeptides isolated from fungal sources. They are characterized by the presence of an unusual amino acid, alpha-aminoisobutyric acid, and a C-terminal hydroxylated amino acid. Peptaibols exhibit antibiotic activity against bacteria and fungi. Their amphipathic nature allows them to self associate into oligomeric ion-channel assemblies which span the width of lipid bilayer membranes. Over 200 peptaibol sequences have been reported to date, which are compiled in the Peptaibol Database at http://www.cryst.bbk.ac.uk/peptaibol. Alignments of these sequences have been carried out in order to define a series of related subfamilies (SFs) with common sequence features thought to be important for channel formation. Crystal structures determined for a number of peptaibols from the various SFs provide the bases both for modelling of the channel structures and for modelling structures of other members of the same SFs. PMID- 11498030 TI - Structural implications for the transformation of the Bacillus thuringiensis delta-endotoxins from water-soluble to membrane-inserted forms. AB - Crystal structures combined with biochemical data show that the delta-endotoxins from Bacillus thuringiensis are structurally poised towards large-scale, irreversible conformational changes that transform them from the soluble protein bound at the cell surface into a membrane-embedded form causing lysis of susceptible insect cells. Cry delta-endotoxins are made of a helix bundle, a beta prism and a beta-sandwich. The conformational change involves an umbrella-like opening between the helix-4,5-hairpin and the remaining helices, and between the helical domain and the two sheet domains. Comparison of Cry1Ac structures with and without the bound receptor ligand GalNAc associates occupation of the high affinity site on the beta-sandwich with an increase of temperature factors in the helical, pore-forming domain, which may indicate how receptor binding could trigger the required major conformational change. The structure of Cyt delta endotoxins indicates that the surface helix hairpins must peel away to expose the beta-strands for membrane attack. Single amino acid substitutions in hinge residues or the core can restore activity following an inhibitory mutation. PMID- 11498031 TI - Assembly of pore proteins on gold electrodes. AB - Protein-protein interactions at the cell surface are important in the activity of bacterial toxins such as colicins. We have developed methods to study these events using tethered lipid bilayers, which can be probed by impedance spectroscopy and surface plasmon resonance. Recently we have attached the receptor proteins directly to gold electrodes and this offers new possibilities for measuring protein-protein interactions on solid supports. PMID- 11498032 TI - Application of electron spin resonance for investigating peptide-lipid interactions, and correlation with thermodynamics. AB - Peptide-lipid interactions can be investigated with spin-labelled lipid probes by using electron spin resonance (ESR) methods that have been developed for studying lipid-protein interactions with both integral and peripheral membrane proteins and also with surface-binding proteins that additionally penetrate the membrane. This approach has the advantage that a direct comparison can be made with the databank of ESR results from the various types of membrane protein. The appropriateness of the peptides as models for membrane proteins, or for their specific segments, can then be assessed. Further, differences in behaviour can be readily identified, as for example in the case of surface-active cytolytic or fusogenic peptides. Comparison with thermodynamic predictions for membrane insertion provides a useful adjunct to the spin-label method. PMID- 11498033 TI - Structural model of a voltage-gated potassium channel based on spectroscopic data. AB - It is estimated that membrane proteins comprise as much as 30% of most genomes. Yet our knowledge of membrane-protein folding is still in its infancy. Consequently, there is a great need for developing approaches that can further advance our understanding of how peptides and proteins interact with membranes and thereby attain their folded structure. An approach that we have been exploring involves dissecting voltage-gated ion channels into simple peptide domains for the purpose of determining their structure in different media using physical techniques. We have synthesized peptides corresponding to the six membrane-spanning segments, as well as the pore domain, of the Shaker channel and characterized their secondary structures. From these studies we have developed a model for the transmembrane structure of the Shaker potassium channel that is constructed from alpha-helices. The hard structural data obtained from these studies lends support to the recent theoretical models of this channel protein that have been developed by others. PMID- 11498034 TI - The energetics of binding of a signal peptide to lipid bilayers: the role of bilayer properties. AB - The interactions between a signal peptide and a variety of lipid bilayers have been studied with direct binding assays, CD and isothermal titration calorimetry. We find that the binding of this peptide is influenced by charges and dipoles located in the bilayer interfacial region, as well as by the presence of cholesterol in the bilayer. These studies show that bilayer compositional and mechanical variations found in different biological membranes can affect the partitioning of peptides into the bilayer. PMID- 11498035 TI - Why and how are peptide-lipid interactions utilized for self defence? AB - Animals defend themselves against invading pathogenic micro-organisms by utilizing cationic anti-microbial peptides, which rapidly kill various micro organisms without exerting toxicity against the host. Physicochemical peptide lipid interactions provide attractive mechanisms for innate immunity. Many of these peptides form amphipathic secondary structures (alpha-helices and beta sheets) which can selectively interact with anionic bacterial membranes by electrostatic interaction. Rapid, peptide-induced membrane permeabilization is an effective mechanism of anti-microbial action. Magainin 2 from frog skin forms a dynamic peptide-lipid supramolecular-complex pore that allows mutually coupled transmembrane transport of ions and lipids. The peptide molecule is internalized upon the disintegration of the pore. Several anti-microbial peptides are known to work synergistically. PMID- 11498036 TI - Membrane properties and amyloid fibril formation of lung surfactant protein C. AB - Pulmonary surfactant is essential for respiration and lung host defence and is composed of 80-90% lipids, mainly dipalmitoylphosphatidylcholine (DPPC). Surfactant protein C (SP-C) constitutes 1-2% of the surfactant mass, and is one of the most hydrophobic peptides yet isolated. SP-C residues 9-34 form an alpha helix with a central poly-valine segment, which perfectly matches the thickness of a fluid DPPC bilayer. The palmitoyl groups linked to Cys-5 and Cys-6 of SP-C increase the capacity of the peptide to promote lipid adsorption at an air/liquid interface, and augment the mechanical stability of SP-C/lipid mixtures. SP-C undergoes alpha-helix-->beta-sheet transition and forms amyloid fibrils. NMR and MS studies show that the poly-valine helix is kinetically stabilized, and that once it unfolds, formation of beta-sheet aggregates is significantly faster than refolding. alpha-Helix unfolding is accelerated after removal of the palmitoyl groups. Secondary structure prediction of SP-C yields beta-strand conformation of the poly-valine part. A database search revealed similar discordance between experimentally determined helices and predicted beta-strands for other amyloid forming proteins, including the prion protein associated with spongiform encephalopathies, and the amyloid-beta (Abeta) peptide associated with Alzheimer's disease. For Abeta and SP-C, removal of the helix/strand discordance by residue replacements abrogates fibril formation in vitro. PMID- 11498037 TI - Connexin mimetic peptides: specific inhibitors of gap-junctional intercellular communication. AB - Intercellular co-operation is a fundamental and widespread feature in tissues and organs. An important mechanism ensuring multicellular homoeostasis involves signalling between cells via gap junctions that directly connect the cytosolic contents of adjacent cells. Cell proliferation and intercellular communication across gap junctions are closely linked, and a number of pathologies in which communication is disrupted are known where connexins, the gap-junctional proteins, are modified. The proteins of gap junctions thus emerge as therapeutic targets inviting the development and exploitation of chemical tools and drugs that specifically influence intercellular communication. Connexin mimetic peptides that correspond to short specific sequences in the two extracellular loops of connexins are a class of benign, specific and reversible inhibitors of gap-junctional communication that have been studied recently in a broad range of cells, tissues and organs. This review summarizes the properties and uses of these short synthetic peptides, and compares their probable mechanism of action with those of a wide range of other less specific traditional gap-junction inhibitors. PMID- 11498038 TI - Tethered-bilayer lipid membranes as a support for membrane-active peptides. AB - An immunosensing device, comprising a lipid membrane incorporating ion channels tethered to the surface of a gold electrode, has been reported [Cornell, Braach Maksvytis, King, Osman, Raguse, Wieczorek and Pace (1997) Nature (London) 387, 580-583]. The present article describes key steps in the assembly of the device and provides further evidence for its proposed sensing mechanism. PMID- 11498039 TI - Membrane-disordering effects of beta-amyloid peptides. AB - beta-Amyloid (Abeta) protein is the major constituent of senile plaques and cerebrovascular deposits characteristic of Alzheimer's disease (AD). The causal relationship between Abeta and AD-specific lesions like neurodegeneration and atrophy is still not known. The present article summarizes our studies indicating that rather low concentrations of Abeta significantly alter the fluidity of cell membranes and subcellular fractions from different tissues and different species including humans, as a possible initial step of its biological effects. Using different fluorescent probes our data show clearly that Abeta peptides specifically disturb the acyl-chain layer of cell membranes in a very distinct fashion. By contrast, membrane properties at the level of the polar heads of the phospholipid bilayer at the interface with membrane proteins are much less affected. PMID- 11498040 TI - Influenza fusion peptides. AB - The 'fusion peptides' of a group of enveloped viruses that includes influenza, paramyxo-, retro-and filo-viruses are N-terminal regions of their membrane fusion proteins generated by cleavage of non-functional precursors. For the influenza membrane fusion protein, haemagglutinin (HA), the three-dimensional structures of precursor HA, cleaved HA and fusion-activated HA show that the fusion peptides are located in different positions in all three forms and adopt different structures. Analyses of mutant HAs with changes in fusion peptide sequence indicate the importance of specific residues for membrane-fusion activity and suggest a structure for the fusion peptide in a fusion-active molecule. PMID- 11498041 TI - Growth and change at Neuron. PMID- 11498042 TI - It takes T to tango. AB - Of three recently cloned T-type voltage-gated calcium channels, alpha(1g) is most likely responsible for burst firing in thalamic relay cells. These neurons burst during various thalamocortical oscillations including absence seizures. In this issue of Neuron, Kim et al. inactivated alpha(1g), and resultant mice were deficient in relay cell bursting and resistant to GABA(B) receptor-dependent absence seizures, suggesting roles for alpha(1g) and relay cell bursting in absences. PMID- 11498043 TI - Making the pain connection. AB - Dorsal root ganglia (DRG) neurons include multiple types of sensory neurons with well-appreciated anatomical and physiological distinctions. In this issue of Neuron, Chen et al. adds to our molecular understanding of these differences by reporting that DRG11, a paired homeodomain transcription factor, is specifically required for the proper development of pain-sensing nociceptive neurons. PMID- 11498044 TI - The perception of inferred action. AB - Our actions, and those of others, are often partly obscured from view. This complicates the sensory inputs that guide motor actions. In this issue of Neuron, Umilita and colleagues demonstrate that "mirror neurons" in ventral premotor cortex respond when monkeys observe hidden, but inferred, actions. PMID- 11498045 TI - TorsinA: movement at many levels. AB - TorsinA is the causative protein in the human neurologic disease early onset torsin dystonia, a movement disorder involving dysfunction in the basal ganglia without apparent neurodegeneration. Most cases result from a dominantly acting three-base pair deletion in the TOR1A gene causing loss of a glutamic acid near the carboxyl terminus of torsinA. Torsins are members of the AAA(+) superfamily of ATPases and are present in all multicellular organisms. Initial studies suggest that torsinA is an ER protein involved in chaperone functions and/or membrane movement. PMID- 11498046 TI - Do you see what I'm saying? Interactions between auditory and visual cortices in cochlear implant users. AB - Primary sensory cortices are generally thought to be devoted to one sensory modality-vision, hearing, or touch, for example. Surprising interactions between these sensory modes have recently been reported. One example demonstrates that people with cochlear implants show increased activity in visual cortex when listening to speech; this may be related to enhanced lipreading ability. PMID- 11498047 TI - Targeting transcription to the neuromuscular synapse. AB - Concomitant with innervation, genes coding for components of the neuromuscular junction become exclusively expressed in subsynaptic nuclei. A six-base pair element, the N box, can confer synapse-specific transcription to the acetylcholine nicotinic receptor delta and epsilon subunit, utrophin, and acetylcholine esterase genes. N box-dependent synaptic expression is stimulated by the nerve-derived signal agrin and the trophic factor neuregulin, which triggers the MAPK and JNK signaling pathways, to ultimately allow activation by the N box binding Ets transcription factor GABP. PMID- 11498048 TI - Sympathetic innervation of lymphoreticular organs is rate limiting for prion neuroinvasion. AB - Transmissible spongiform encephalopathies are commonly propagated by extracerebral inoculation of the infectious agent. Indirect evidence suggests that entry into the central nervous system occurs via the peripheral nervous system. Here we have investigated the role of the sympathetic nervous system in prion neuroinvasion. Following intraperitoneal prion inoculation, chemical or immunological sympathectomy delayed or prevented scrapie. Prion titers in spinal cords were drastically reduced at early time points after inoculation. Instead, keratin 14-NGF transgenic mice, whose lymphoid organs are hyperinnervated by sympathetic nerves, showed reduction in scrapie incubation time and, unexpectedly, much higher titers of prion infectivity in spleens. We conclude that sympathetic innervation of lymphoid organs is rate limiting for prion neuroinvasion and that splenic sympathetic nerves may act as extracerebral prion reservoirs. PMID- 11498049 TI - Lack of the burst firing of thalamocortical relay neurons and resistance to absence seizures in mice lacking alpha(1G) T-type Ca(2+) channels. AB - T-type Ca(2+) currents have been proposed to be involved in the genesis of spike and-wave discharges, a sign of absence seizures, but direct evidence in vivo to support this hypothesis has been lacking. To address this question, we generated a null mutation of the alpha(1G) subunit of T-type Ca(2+) channels. The thalamocortical relay neurons of the alpha(1G)-deficient mice lacked the burst mode firing of action potentials, whereas they showed the normal pattern of tonic mode firing. The alpha(1G)-deficient thalamus was specifically resistant to the generation of spike-and-wave discharges in response to GABA(B) receptor activation. Thus, the modulation of the intrinsic firing pattern mediated by alpha(1G) T-type Ca(2+) channels plays a critical role in the genesis of absence seizures in the thalamocortical pathway. PMID- 11498050 TI - Epilepsy, hyperalgesia, impaired memory, and loss of pre- and postsynaptic GABA(B) responses in mice lacking GABA(B(1)). AB - GABA(B) (gamma-aminobutyric acid type B) receptors are important for keeping neuronal excitability under control. Cloned GABA(B) receptors do not show the expected pharmacological diversity of native receptors and it is unknown whether they contribute to pre- as well as postsynaptic functions. Here, we demonstrate that Balb/c mice lacking the GABA(B(1)) subunit are viable, exhibit spontaneous seizures, hyperalgesia, hyperlocomotor activity, and memory impairment. Upon GABA(B) agonist application, null mutant mice show neither the typical muscle relaxation, hypothermia, or delta EEG waves. These behavioral findings are paralleled by a loss of all biochemical and electrophysiological GABA(B) responses in null mutant mice. This demonstrates that GABA(B(1)) is an essential component of pre- and postsynaptic GABA(B) receptors and casts doubt on the existence of proposed receptor subtypes. PMID- 11498051 TI - The paired homeodomain protein DRG11 is required for the projection of cutaneous sensory afferent fibers to the dorsal spinal cord. AB - Cutaneous sensory neurons that detect noxious stimuli project to the dorsal horn of the spinal cord, while those innervating muscle stretch receptors project to the ventral horn. DRG11, a paired homeodomain transcription factor, is expressed in both the developing dorsal horn and in sensory neurons, but not in the ventral spinal cord. Mouse embryos deficient in DRG11 display abnormalities in the spatio temporal patterning of cutaneous sensory afferent fiber projections to the dorsal, but not the ventral spinal cord, as well as defects in dorsal horn morphogenesis. These early developmental abnormalities lead, in adults, to significantly attenuated sensitivity to noxious stimuli. In contrast, locomotion and sensori-motor functions appear normal. Drg11 is thus required for the formation of spatio-temporally appropriate projections from nociceptive sensory neurons to their central targets in the dorsal horn of the spinal cord. PMID- 11498052 TI - Redox modulation of T-type calcium channels in rat peripheral nociceptors. AB - Although T-type calcium channels were first described in sensory neurons, their function in sensory processing remains unclear. In isolated rat sensory neurons, we show that redox agents modulate T currents but not other voltage- and ligand gated channels thought to mediate pain sensitivity. Similarly, redox agents modulate currents through Ca(v)3.2 recombinant channels. When injected into peripheral receptive fields, reducing agents, including the endogenous amino acid L-cysteine, induce thermal hyperalgesia. This hyperalgesia is blocked by the oxidizing agent 5,5'-dithio-bis-(2-nitrobenzoic acid) (DTNB) and the T channel antagonist mibefradil. DTNB alone and in combination with mibefradil induces thermal analgesia. Likewise, L-cysteine induces mechanical DTNB-sensitive hyperalgesia in peripheral receptive fields. These data strongly suggest a role for T channels in peripheral nociception. Redox sites on T channels in peripheral nociceptors could be important targets for agents that modify pain perception. PMID- 11498053 TI - Multiple phosphorylation of rhodopsin and the in vivo chemistry underlying rod photoreceptor dark adaptation. AB - Dark adaptation requires timely deactivation of phototransduction and efficient regeneration of visual pigment. No previous study has directly compared the kinetics of dark adaptation with rates of the various chemical reactions that influence it. To accomplish this, we developed a novel rapid-quench/mass spectrometry-based method to establish the initial kinetics and site specificity of light-stimulated rhodopsin phosphorylation in mouse retinas. We also measured phosphorylation and dephosphorylation, regeneration of rhodopsin, and reduction of all-trans retinal all under identical in vivo conditions. Dark adaptation was monitored by electroretinography. We found that rhodopsin is multiply phosphorylated and then dephosphorylated in an ordered fashion following exposure to light. Initially during dark adaptation, transduction activity wanes as multiple phosphates accumulate. Thereafter, full recovery of photosensitivity coincides with regeneration and dephosphorylation of rhodopsin. PMID- 11498054 TI - Functional assembly of AMPA and kainate receptors is mediated by several discrete protein-protein interactions. AB - Functional heterogeneity of ionotropic glutamate receptors arises not only from the existence of many subunits and isoforms, but also from combinatorial assembly creating channels with distinct properties. This heteromerization is subtype restricted and thought to be determined exclusively by the proximal extracellular N-terminal domain of the subunits. However, using functional assays for heteromer formation, we show that, besides the N-terminal domain, the membrane sector and the C-terminal part of S2 are critical determinants for the formation of functional channels. Our results are compatible with a model where the N-terminal domain only mediates the initial subunit associations into dimers, whereas for the assembly of the full functional tetramer, compatibility of the other regions is required. PMID- 11498055 TI - Regulation of dendritic spine morphology and synaptic function by Shank and Homer. AB - The Shank family of proteins interacts with NMDA receptor and metabotropic glutamate receptor complexes in the postsynaptic density (PSD). Targeted to the PSD by a PDZ-dependent mechanism, Shank promotes the maturation of dendritic spines and the enlargement of spine heads via its ability to recruit Homer to postsynaptic sites. Shank and Homer cooperate to induce accumulation of IP3 receptors in dendritic spines and formation of putative multisynapse spines. In addition, postsynaptic expression of Shank enhances presynaptic function, as measured by increased minifrequency and FM4-64 uptake. These data suggest a central role for the Shank scaffold in the structural and functional organization of the dendritic spine and synaptic junction. PMID- 11498056 TI - Timing and location of nicotinic activity enhances or depresses hippocampal synaptic plasticity. AB - This study reveals mechanisms in the mouse hippocampus that may underlie nicotinic influences on attention, memory, and cognition. Induction of synaptic plasticity, arising via generally accepted mechanisms, is modulated by nicotinic acetylcholine receptors. Properly timed nicotinic activity at pyramidal neurons boosted the induction of long-term potentiation via presynaptic and postsynaptic pathways. On the other hand, nicotinic activity on interneurons inhibited nearby pyramidal neurons and thereby prevented or diminished the induction of synaptic potentiation. The synaptic modulation was dependent on the location and timing of the nicotinic activity. Loss of these synaptic mechanisms may contribute to the cognitive deficits experienced during Alzheimer's diseases, which is associated with a loss of cholinergic projections and with a decrease in the number of nicotinic receptors. PMID- 11498057 TI - Molecular mechanisms underlying a unique intermediate phase of memory in aplysia. AB - Short- and long-term synaptic facilitation induced by serotonin at Aplysia sensory-motor (SN-MN) synapses has been widely used as a cellular model of short- and long-term memory for sensitization. In recent years, a distinct intermediate phase of synaptic facilitation (ITF) has been described at SN-MN synapses. Here, we identify a novel intermediate phase of behavioral memory (ITM) for sensitization in Aplysia and demonstrate that it shares the temporal and mechanistic features of ITF in the intact CNS: (1) it declines completely prior to the onset of LTM, (2) its induction requires protein but not RNA synthesis, and (3) its expression requires the persistent activation of protein kinase A. Thus, in Aplysia, the same temporal and molecular characteristics that distinguish ITF from other phases of synaptic plasticity distinguish ITM from other phases of behavioral memory. PMID- 11498058 TI - I know what you are doing. a neurophysiological study. AB - In the ventral premotor cortex of the macaque monkey, there are neurons that discharge both during the execution of hand actions and during the observation of the same actions made by others (mirror neurons). In the present study, we show that a subset of mirror neurons becomes active during action presentation and also when the final part of the action, crucial in triggering the response in full vision, is hidden and can therefore only be inferred. This implies that the motor representation of an action performed by others can be internally generated in the observer's premotor cortex, even when a visual description of the action is lacking. The present findings support the hypothesis that mirror neuron activation could be at the basis of action recognition. PMID- 11498060 TI - Medical therapy or coronary artery bypass graft surgery for chronic stable angina: an update using decision analysis. AB - PURPOSE: Randomized trials comparing medical and surgical therapies for the treatment of chronic stable angina were completed in the early 1980s. Therapies developed since then have decreased mortality and myocardial infarction rates from coronary artery disease. Using decision analysis and incorporating current recommendations for treatment, we simulated a trial comparing coronary artery bypass graft surgery and medical therapy. METHODS: A Markov decision analysis model was constructed to compare the 5-year and 10-year outcomes of a simulated trial of medical therapy versus bypass surgery for stable chronic angina. Baseline data were obtained from a meta-analysis of trials comparing the two treatments. Data on risk reduction from contemporary therapies were obtained from randomized trials and meta-analyses. RESULTS: All subgroups experienced modest gains in survival with current therapies. At 5 years, the survival rate was 90% in the medical group (an absolute gain of 6%) and 94% in the surgical group (an absolute gain of 4%). Similar results were obtained for patients with triple vessel disease. Among patients with a low ejection fraction, the 5-year survival rate was 85% for medical patients and 92% for surgical patients. Sensitivity analyses did not substantially affect the conclusions. CONCLUSION: Advances in the treatment of chronic stable angina have improved the outcome both for patients treated initially with surgery and for those treated initially with medical therapy. The improvements were of similar magnitude in both groups, so the fundamental conclusions of the bypass trials are unchanged. PMID- 11498061 TI - Pathophysiologic determinants of third heart sounds: a prospective clinical and Doppler echocardiographic study. AB - PURPOSE: We sought to determine the importance of a third heart sound (S(3)) and its relation to hemodynamic and valvular dysfunction. SUBJECTS AND METHODS: We prospectively enrolled 580 patients who had isolated valvular regurgitation (mitral, n = 299; aortic, n = 121) or primary left ventricular dysfunction with or without functional mitral regurgitation (n = 160). We analyzed the associations between the clinical finding of an audible S(3) (as noted in routine clinical practice by internal medicine physicians) and hemodynamic alterations measured by comprehensive quantitative Doppler echocardiography. RESULTS: S(3) was more prevalent in patients with primary left ventricular dysfunction (46%, n = 73) than in organic mitral (16%, n = 47) or aortic (12%, n = 14) regurgitation (P <0.001). Patients with an S(3) were more likely to have class III-IV symptoms (55% [74 of 137] vs. 18% [80 of 443] of those without an S(3), P <0.001) and had a higher mean [+/- SD] pulmonary pressure (55 +/- 15 vs. 41 +/- 11 mm Hg, P <0.001). An S(3) was also related to a higher early filling velocity due to a greater filling volume, restrictive filling, or both. An S(3) was a marker of severe regurgitation (regurgitant fraction > or =40%) in patients with primary left ventricular dysfunction (odds ratio [OR] = 2.4; 95% confidence interval [CI]: 1.1 to 5.5), mitral regurgitation (OR = 17; 95% CI: 5.8 to 52), and aortic regurgitation (OR = 7.1; 95% CI: 1.8-28). An S(3) was also associated with restrictive filling in primary left ventricular dysfunction (OR = 3.0; 95% CI, 1.6 to 5.9), marked dilatation in mitral regurgitation (OR = 20; 95% CI: 6.8 to 58), and an ejection fraction (<50%) in aortic regurgitation (OR = 19; 95% CI: 6.0 to 62). CONCLUSION: An audible S(3) is an important clinical finding, indicating severe hemodynamic alterations, and should lead to a comprehensive assessment and consideration of vigorous medical or surgical treatment. PMID- 11498062 TI - A randomized trial of zinc nasal spray for the treatment of upper respiratory illness in adults. AB - PURPOSE: We performed a randomized, double-blind, placebo-controlled trial to evaluate the efficacy of 0.12% zinc sulfate nasal spray for reducing the duration and severity of acute upper respiratory infections. SUBJECTS AND METHODS: Patients with acute onset of upper respiratory illness of less than 24 hours' duration were eligible for the study. A nasopharyngeal swab was obtained at the time of enrollment for viral culture. Participants were randomly assigned to receive either 0.12% zinc sulfate or isotonic placebo spray. The medication was administered as two inhalations in each nostril four times a day. Each patient completed a diary card twice a day to record oral temperature, symptoms, and adverse effects. Symptoms were scored as absent (0), mild (1), moderate (2), or severe (3). RESULTS: One hundred eighty-five subjects volunteered to participate, and 160 met the criteria for enrollment. The median duration of all symptoms was 7 days in both groups (P = 0.45), and the median duration of nasal symptoms was 6 days in both groups (P= 0.12). After adjustment for baseline differences in severity, patients receiving zinc had a significant reduction in the total symptom score (P= 0.02) and the nasal symptom score (P= 0.02) on day 1, but not on any of the other days. Adverse effects were mild and had no significant association with the use of zinc. A respiratory virus was identified in 9 of the 160 participants; 6 of these were rhinovirus. CONCLUSION: A low concentration of zinc sulfate nasal spray had no effect on the duration of the common cold. PMID- 11498063 TI - Eosinophilic meningitis caused by Angiostrongylus cantonensis: report of 17 cases. AB - PURPOSE: To describe two outbreaks of Angiostrongylus cantonensis infection that occurred in Kaohsiung, Taiwan, during 1998 and 1999, and to characterize the source of the outbreaks and the clinical manifestations of the disease. SUBJECTS AND METHODS: We performed a retrospective cohort study among Thai laborers with eosinophilic meningitis who ate raw snails (Ampullarium canaliculatus), as well as an environmental surveillance of larvae in snails. RESULTS: We enrolled 17 Thai laborers in whom severe headache and eosinophilia developed within 4 to 23 days after eating raw snails. Twelve (71%) developed eosinophilic meningitis. Third-stage larvae were found in the cerebrospinal fluids of 2 patients and in all 12 tested snails. Specific antibodies to A. cantonensis were detected in serum from 16 of the patients and in cerebrospinal fluid from 5 of the patients. Central nervous system manifestations included headache (n = 17 [100%]), fever (n = 11 [65%]), Brudzinski's sign/stiff neck (n = 11 [65%]), hyperesthesia (n = 3 [18%]), cranial nerve palsy (n = 2 [12%]), diplopia (n = 2 [12%]), and ataxia (n = 1 [6%]). Laboratory findings included peripheral eosinophilia (n = 15 [88%]) and cerebrospinal fluid eosinophilia (n = 12 [71%]); elevated immunoglobulin (Ig) E levels (n = 13 [100%]); and transient increases in white blood cell count (n = 7 [41%]) and in serum levels of creatine kinase (n = 7 [41%]), transaminase (n = 3 [18%]), and lactate dehydrogenase (n = 2 [12%]). The severity of illness and eosinophilia were correlated with the number of ingested snails. Meningeal and basal ganglion enhancement was noted on magnetic resonance imaging in several patients. Treatment with mebendazole combined with glucocorticosteroids appeared to shorten the course of the infection, but not the number of relapses. The eosinophil count fell to normal within 3 months, but IgE levels remained elevated for as long as 6 months. All patients recovered with minimal neurologic sequelae. CONCLUSION: Eosinophilic meningitis caused by A. cantonensis should be considered in patients who have headache or central nervous system manifestations after eating raw snails. PMID- 11498064 TI - Nonconvulsive status epilepticus associated with cephalosporins in patients with renal failure. AB - PURPOSE: Nonconvulsive status epilepticus is an unusual complication of cephalosporin therapy, with only a few isolated cases reported. SUBJECTS AND METHODS: We reviewed the clinical and electroencephalographic (EEG) characteristics of 10 patients with renal failure in whom developed alteration of consciousness without convulsions associated with continuous epileptiform EEG activity while being treated with cephalosporins. RESULTS: Nonconvulsive status epilepticus developed in 5 men and 5 women, with a mean (+/- SD) age of 69 +/- 14 years, while receiving intravenous cephalosporins (ceftriaxone, 2 patients; ceftazidime, 2; and cefepime, 6). All patients had renal failure; 1 also had hepatic failure. Patients presented with progressive disorientation or agitation, sometimes associated with mild facial or limb myoclonus, that had begun 1 to 10 days (mean, 5 +/- 2 days) after starting cephalosporin treatment. The EEG showed continuous or intermittent bursts of generalized, high-voltage, 1 to 2 Hz sharp wave activity or sharp and slow wave activity that resembled, but could be differentiated from, the triphasic waves seen in metabolic encephalopathies. Intravenous clonazepam suppressed the epileptiform activity completely in 5 patients and partially in the other 5. Cephalosporins were withdrawn, and antiepileptic therapy was started for all patients. All patients improved, 2 in less than 24 hours and the remainder within 2 to 7 days. CONCLUSIONS: Cephalosporins can cause nonconvulsive status epilepticus in patients with renal failure. The clinical picture is difficult to differentiate from a that of metabolic encephalopathy unless an EEG is obtained. Physicians should be aware of this potentially dangerous complication. PMID- 11498065 TI - Excess mortality in women with hospital-acquired bloodstream infection. AB - PURPOSE: We examined the outcomes of bloodstream infection in men and in women and whether any sex-related differences were explained by underlying disorders, severity of disease, or clinical management. SUBJECTS AND METHODS: Using a prospectively collected database, we compared in-hospital mortality in men and women. We used multivariable logistic regression analysis to test whether sex related differences could be due to potential confounders. RESULTS: Of 4250 patients with bloodstream infections, 1750 (41%) had hospital-acquired infections. The overall case fatality was 31% (625 of 2032) in women and 29% (631 of 2218, P = 0.1) in men. However, 43% (325/758) of the women with hospital acquired infections died, compared with 33% (327/992) of the men (P = 0.0001). In a multivariate analysis, female sex was associated with greater mortality in patients with hospital-acquired infections (odds ratio = 1.7; 95% confidence interval: 1.1 to 2.6). The excess mortality in women was mainly seen in patients with major underlying disorders (fatality rate of 45% [234 of 525] in women vs. 32% in men [234 of 743, P = 0.0001). CONCLUSIONS: Mortality in women with hospital-acquired bloodstream infections is substantially greater than in men. The excess mortality was concentrated in women with severe underlying disorders, suggesting that sepsis might have accentuated differences in the outcome of underlying disorders in women. PMID- 11498066 TI - Oral cobalamin therapy for the treatment of patients with food-cobalamin malabsorption. AB - BACKGROUND: The standard treatment for cobalamin (vitamin B(12)) deficiency involves regular intramuscular cobalamin injection. It has been suggested that oral cobalamin therapy may be effective for treating patients who have food cobalamin malabsorption. SUBJECTS AND METHODS: We prospectively studied 10 patients with cobalamin deficiency and well-established food-cobalamin malabsorption who received 3000 microg or 5000 microg of oral crystalline cyanocobalamin once a week for at least 3 months. Complete blood counts and serum cobalamin, homocysteine, and folate levels were determined at baseline and after 3 months of treatment. Patients were reexamined after 6 months. RESULTS: After 3 months of treatment, all patients had increased hemoglobin levels (mean increase, 1.9 g/dL; 95% confidence interval: 0.9 to 3.9 g/dL;P <0.01 compared with baseline) and decreased erythrocyte cell volume (mean decrease, 7.8 fL; 95% confidence interval: 0.9 to 16.5 fL;P<0.001). However, 2 patients had only minor, if any, responses. Serum cobalamin levels were increased in all 8 patients in whom it was measured. CONCLUSION: Our findings suggest that moderate doses of crystalline cyanocobalamin given orally may be an effective treatment for food cobalamin malabsorption. PMID- 11498067 TI - Low-molecular-weight heparin versus warfarin for secondary prophylaxis of venous thromboembolism: a cost-effectiveness analysis. AB - PURPOSE: To compare the cost effectiveness of low-molecular-weight heparin with that of oral anticoagulants in preventing recurrences after an episode of venous thromboembolism. METHODS: A decision tree was used to assess the cost and the expected quality-adjusted years of life (QALY) after treatment with either low molecular-weight heparin or warfarin, based on pooled data from six published trials. Preferences were elicited with a modified time trade-off method in a sample of patients attending an anticoagulation clinic. RESULTS: Compared with warfarin, low-molecular-weight heparin significantly decreased the rate of minor bleeding (odds ratio [OR] = 0.24; 95% confidence interval [CI]: 0.14 to 0.43) but not recurrent deep vein thromboses (OR = 0.77; 95% CI: 0.43 to 1.35). Patients' preference for warfarin (0.988, on a 0 to 1 scale) was lower than that for low molecular-weight heparin (0.992), but the difference was not statistically significant. A Monte Carlo analysis estimated that low-molecular-weight heparin saved an average of 13 quality-adjusted days compared with warfarin, at a cost of $6,583 per QALY (95% CI: $5,525 to $7,625) based on costs in Italy and $28,231 per QALY (95% CI: $20,872 to $36,773) based on costs in the United States. When we included rebound recurrences after interruption of therapy, which were more common with low-molecular-weight heparin, treatment with low-molecular-weight heparin cost $53,166 per QALY in Italy and $177,166 per QALY in the United States. CONCLUSIONS: Low-molecular-weight heparin might be a cost-effective drug for secondary prophylaxis of venous thromboembolism, especially in patients at high risk of recurrence and where the drug's cost is lower. The apparent increase in recurrence after interruption of therapy needs to be investigated more thoroughly before low-molecular-weight heparin can be recommended routinely. PMID- 11498068 TI - Cost effectiveness of human papillomavirus testing to augment cervical cancer screening in women infected with the human immunodeficiency virus. AB - PURPOSE: To determine the cost effectiveness of incorporating molecular testing for high-risk types of human papillomavirus into a cervical cancer screening program for women infected with the human immunodeficiency virus (HIV). SUBJECTS AND METHODS: We developed a Markov model to simulate the natural history of cervical cancer precursor lesions in HIV-infected women. Probabilities of progression and regression of cervical lesions were conditional on transient or persistent infection with human papillomavirus, as well as stage of HIV and effectiveness of antiretroviral therapy. Incorporating data from prospective cohort studies, national databases, and published literature, the model was used to calculate quality-adjusted life expectancy, life expectancy, lifetime costs, and incremental cost-effectiveness ratios for two main strategies: targeted screening-human papillomavirus testing is added to the initial two cervical cytology smears obtained after an HIV diagnosis and subsequent screening intervals are modified based on the test results; and universal screening-no testing for human papillomavirus is performed, and a single cytology screening interval is applied to all women. RESULTS: In HIV-infected women on anti retroviral therapy, a targeted screening strategy in which cervical cytology screening was conducted every 6 months for women with detected human papillomavirus DNA, and annually for all others, cost $10,000 to $14,000 per quality-adjusted life year gained compared with no screening. A universal screening strategy consisting of annual cervical cytology for all women was 15% less effective and had a less attractive cost-effectiveness ratio. Targeted screening remained economically attractive in multiple sensitivity analyses, although when the overall incidence of cervical cancer precursor lesions was lowered by 75%, the screening interval for women with detected human papillomavirus DNA could be widened to 1 year. CONCLUSIONS: Adding human papillomavirus testing to the two cervical cytology smears obtained in the year after an HIV diagnosis, and modifying subsequent cytology screening intervals based on the results, appears to be an effective and cost-effective modification to current recommendations for annual cytology screening in HIV-infected women. PMID- 11498069 TI - Methemoglobinemia complicating topical lidocaine used during endoscopic procedures. PMID- 11498070 TI - Can mathematical modeling remove the tarnish on old clinical trials? PMID- 11498071 TI - The clinical meaning of the third heart sound. PMID- 11498072 TI - Treatment of venous thromboembolism using only low-molecular-weight heparin? PMID- 11498074 TI - Predicting and preventing physician burnout: results from the United States and the Netherlands. PMID- 11498075 TI - Plasma and RBCs antioxidant status in occupational male pesticide sprayers. AB - A total of forty-one (n=41) male, healthy agricultural sprayers, exposed to pesticides for 5 years, were compared with twenty one (n=21) controls matched for age and economic status with respect to free radical generation, lipid peroxidation, antioxidant status and concentration of cellular enzymes were determined. Significantly increased TBARS (thiobarbituric acid reactive substances) were observed (P<0.001) in sprayer populations when compared to controls. The concentration of antioxidants such as glutathione (GSH), alpha tocopherol, ascorbic acid and ceruloplasmin were significantly altered when compared to controls, and the activities of antioxidant enzymes were remarkably elevated (P<0.001) in sprayer populations, when compared to controls. PMID- 11498076 TI - Serum concentration and circadian profiles of cathepsins B, H and L, and their inhibitors, stefins A and B, in asthma. AB - BACKGROUND: In order to determine the effect of asthma on serum concentrations of cathepsins B, H and L, and stefins A and B, the circadian and concentration profiles were followed in steroid-independent and steroid-dependent asthmatics before and after 1-week treatment with methylprednisolone and cyclosporin A. METHODS: Serum samples were taken at 4-h intervals throughout a 24-h period. Cathepsin and stefin concentrations were assayed using specific ELISAs. Data were analysed by one-way ANOVA and least squares fit of 24-h cosine. RESULTS: Temporal analysis of these proteins revealed little or no significant changes with time over a 24-h period. In comparison to normal sera, cathepsin H concentrations were elevated in all asthmatic patients, concentrations of both stefins were decreased in steroid-independent asthmatics, and stefin A concentrations were increased in steroid-dependent asthmatics before therapy. The effect of methylprednisolone treatment was demonstrated on decreased cathepsin B and increased cathepsin L concentrations in post-therapy serum samples. On the other hand, cyclosporin A treatment led to increased concentrations of cathepsins H and L. However, concentrations of stefins A and B were unaffected. CONCLUSIONS: This study associated alterations in balance of serum cysteine proteinases and their inhibitors in asthmatic patients, which has raised the possibility of their involvement in asthma pathogenesis. Validated rhythms of cathepsins and stefins in asthmatic sera exhibited temporal differences, which are too small to influence the time of sampling for their quantitative measurement over the course of a day. PMID- 11498077 TI - Basal production of pentane in expired gas from healthy humans. AB - BACKGROUND: Pentane in exhaled gas is often used as an index of lipoperoxidation, but today, there is no standardization for its measurement. In this study, with our technical experience, we determined basal production of pentane in healthy subjects, and we evaluated variability of pentane flow 1 month later. METHODS: 18 subjects inhaled hydrocarbon-free air (HCFA) in order to realize a lung washout. Ambient air and three samples (at T0, T10, T30 min) of expired gas were concentrated using a "trap-and-purge" procedure. For the analysis of pentane, an Al(2)O(3)/KCl plot column contained in a gas chromatograph equipped with a flame ionization detector was used. RESULTS: After 10 min of washout, mean (+/-SD) exhalation rate of pentane was 1+/-0.6 pmol min(-1) kg(-1). After 30 min of washout, mean (+/-SD) exhalation rate of pentane was 0.7+/-0.5 pmol min(-1) kg( 1). No significant difference in pentane flow was shown 1 month later for eight subjects who repeated the protocol. CONCLUSION: With our results and data of the literature, exhalation rates of pentane from healthy adults appear to range between 0.3 and 2 pmol min(-1) kg(-1). The variability of pentane flow 1 month later seems not very important. PMID- 11498078 TI - A malignant phenotype of hypertrophic cardiomyopathy caused by Arg719Gln cardiac beta-myosin heavy-chain mutation in a Chinese family. AB - Mutations of the cardiac beta-myosin heavy-chain (beta-MHC) gene cause hypertrophic cardiomyopathy (HCM). Recent genotype-phenotype correlation studies have shown that mutations carry prognostic significance. We studied five unrelated Chinese families with hypertrophic cardiomyopathy. Exons 3-27 and 40 of the beta-MHC gene were screened with both the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method and the cycle sequencing of the PCR products. A previously reported heterozygous mutation Arg719Gln (arginine ->glutamine in codon 719) in exon 19 was found in one family. The proband is a 30 year-old female diagnosed at age of 25 years when she presented with symptoms of chest pain, palpitations, and frequent incidents of dizziness and syncope. A two dimensional echocardiogram showed moderate asymmetrical septal hypertrophy with left atrial enlargement. There was no obstruction of the left ventricular outflow tract (LVOT). The patient also developed atrial fibrillation. The proband's mother and one of her sisters had similar clinical manifestations and both died suddenly at the age of 38 years. In addition, two silent nucleotide substitutions (ACT63ACC, TTT244TTC) in the cardiac beta-MHC gene were identified in the other four families. These synonymous mutations did not cosegregate with the disease in the families and they were also present in the 60 healthy and age-matched control subjects. Of the five families studied, we did not find any missense mutation in the remaining four families. The missense mutation Arg719Gln found in the Chinese family is associated with a malignant phenotype of severe clinical symptoms and poor survival prognosis. This mutation also causes atrial enlargement and atrial fibrillation. Our study provides further evidence that the mutation, which alters the charge of the myosin heavy chain, is associated with a serious clinical outcome. PMID- 11498079 TI - Development of a bloodspot assay for insulin. AB - BACKGROUND: Bloodspot assays for hormones have advantages of ease of collection/transportation, minimal invasiveness and small blood volume. Such an approach would be of use in epidemiological studies of insulin resistance in children and neonates. METHODS: A two-site chemiluminescent immunoassay for the quantitation of insulin in bloodspots dried on filter paper, based on modifications of a commercially available kit, was used. Analysis was carried out on two 3-mm diameter discs punched out from whole blood standards and test samples. RESULTS: The detection limit of the assay was 5.9 pmol/l of whole blood, with mean intra-assay CV and mean inter-assay CV being <15% above 24 pmol/l. The mean recovery of added insulin was 83%. Comparison of paired whole bloodspots and serum samples collected simultaneously gave a correlation of 0.89. The bloodspot insulin concentrations were stable in excess of 2 months when stored at -20 degrees C and were not susceptible to degradation after four freeze-thaw cycles. CONCLUSIONS: This simple and convenient method is suitable for the measurement of insulin in small volumes of blood collected on filter paper cards and can be applied to epidemiological studies of insulin resistance. PMID- 11498080 TI - Chemiluminescence in inflammatory bowel disease patients: a parameter of inflammatory activity. AB - BACKGROUND: Reactive oxygen species (ROS) are produced in excess in the inflamed mucosa and peripheral blood of patients with inflammatory bowel disease. These species have emerged as a common pathway of tissue injury in a wide variety of inflammatory and other disease processes. The present study was conducted to assess ROS production and to correlate this with parameters of inflammatory activity. METHODS: In 25 patients with Crohn's disease (CD), 20 patients with ulcerative colitis (UC) and 65 age- and sex-matched healthy volunteers ROS production was measured using the whole blood luminol enhanced chemiluminescence assay (LECA). Disease activity was assessed using the Crohn's disease activity index and the Ulcerative Colitis Symptoms Score (UCSS) for CD and UC, respectively. Furthermore, the effect of various scavengers, enzymes and enzyme inhibitors on LECA was studied to assess the contribution of different ROS. RESULTS: LECA was significantly higher in CD and UC patients compared with healthy controls (7.1+/-4.7 and 9.8+/-6 vs. 5.2+/-2.8 x 10(3) counts per minute (cpm), p<0.05 and <0.001). In CD, relative LECA (patient/control) was correlated with the Crohn's disease activity index and C-reactive protein (CRP) (r=0.54, p=0.001 and r=0.51, p=0.01). In UC, CRP but not LECA was correlated with the Ulcerative Colitis Symptoms Score (C-reactive protein: r=0.42, p=0.01). Addition of azide, superoxide dismutase, deferoxamine and dimethylthiourea resulted in a decrease of LECA values. CONCLUSION: Whole blood LECA is increased in patients with CD and UC. This parameter is correlated with disease activity in CD. The observed chemiluminescence is probably due to generation of superoxide and the hydroxyl radical. PMID- 11498081 TI - Conformational state and receptor recognition of the C-terminal domain of human alpha(2)-macroglobulin after dissociation into half-molecules. AB - BACKGROUND: Dissociation of native human alpha(2)-macroglobulin (alpha(2)M) by sodium thiocyanate generates stable half-molecules with intact thiol esters. Significant conformational changes occur by the dissociation, which are similar to those occurring by transformation from native to methylamine-treated alpha(2) macroglobulin. METHODS: The conformational state of the receptor-binding domain of the half-molecules was investigated by receptor binding and clearance studies, and by use of a panel of 11 monoclonal antibodies (mAbs) specific for the 18-kDa C-terminal receptor-binding fragment of alpha(2)-macroglobulin. RESULTS: The half molecules simultaneously express epitopes specific for native, as well as epitopes specific for transformed alpha(2)-macroglobulin. While it is possible to immunochemically discriminate between the different forms of tetrameric protein, the half-molecules retain a conformational state with no observed conformational changes in the C-terminal domain following cleavage of thiol esters or bait regions. The in vivo clearance rate in mice was consequently significantly slower for the half-molecules than for the tetrameric receptor-recognized forms of alpha(2)-macroglobulin. Furthermore, half-molecules demonstrate lower affinity for binding to mouse macrophages than methylamine-treated tetrameric alpha(2) macroglobulin in competition studies. CONCLUSIONS: It is suggested that contact zones are functionally important for mediating conformational switches, which result in trapping and exposure of the receptor-binding sites. PMID- 11498082 TI - Evaluation and comparison of therapeutic monitoring of whole-blood levels of cyclosporin A and its metabolites in renal transplantation by HPLC and RIA methods. AB - BACKGROUND: The aim of the work was to evaluate the possibility to estimate the level of cyclosporin A (CyA) metabolites as the difference of radioimmunoassay (RIA) non-specific and RIA specific methods. METHODS: Blood samples of renal transplant patients were analyzed by three different methods: RIA specific method (CYCLO-Trac, DiaSorin, USA) (RIA(SP)), RIA non-specific method (Immunotech, Czech Republic) (RIA(NS)), and high performance liquid chromatography (HPLC) method. RESULTS: Although values obtained by RIA(SP) correlated well those obtained by HPLC (RIA(SP)=0.995.HPLC+9.68; r(2)=0.962, n=448), the results of HPLC methods were lower by 8%. The values obtained by RIA(NS) were 2.57 times higher than the values obtained by RIA(SP) (RIA(SP)=0.356RIA(NS); r(2)=0.713, n=448). The ratio (CyA+CyA metabolites)/(CyA) calculated as the ratio RIA(NS)/RIA(SP) values for 42 renal transplant patients was relatively stable for each particular patient. The sum of selected CyA metabolites (M1+M17+M21) measured by HPLC correlated well with that estimated from the difference of RIA(NS)-RIA(SP): HPLC(metab)=0.921.(RIA(NS)-RIA(SP))+21.3; (r(2)=0.746, n=448). CONCLUSION: The combination of both the specific and non-specific methods for the determination of CyA presents an improved means for the TDM of CyA and CyA metabolites in renal transplant patients. Moreover, a combination of both methods can help to elucidate some unexpected events, such as the persistence of high cyclosporin blood levels. PMID- 11498083 TI - Dynamics of brain-derived proteins in cerebrospinal fluid. AB - BACKGROUND: The recent theory of blood-cerebrospinal fluid (CSF) barrier function and dysfunction connects molecular flux and CSF flow rate. A reduced CSF flow rate is sufficient to account for the observed hyperbolic relation between different blood-derived protein concentrations in CSF in cases of a blood-CSF barrier dysfunction. METHODS: The dynamics of brain-derived proteins in CSF are investigated with reference to the CSF flow rate measured by CSF/serum albumin concentration quotient. RESULTS: Proteins from neurons or glial cells, tau protein, neuron-specific enolase, S-100 protein, all enter CSF primarily in the ventricular and cisternal space. Their concentration between normal ventricular and lumbar CSF is decreasing (in contrast to blood-derived proteins), and in the case of pathologically decreasing CSF flow rate, the concentration in lumbar CSF remains invariantly constant. Concentrations of the primarily leptomeningeal proteins, beta-trace protein and cystatin C, increase between normal ventricular and lumbar CSF, and in the case of pathologically decreased CSF flow rate they increase linearly in lumbar CSF (concentrations of blood-derived proteins increase non-linearly). CONCLUSIONS: A satisfactory physiological explanation can now be given for the dynamics of proteins in CSF consisting of both brain- and blood-derived fractions (transthyretin, soluble intercellular adhesion molecule (s-ICAM)), as well as the disputed decrease of leptomeningeal protein concentrations (beta-trace protein, cystatin C) in cases of bacterial meningitis is also explained. The biophysical treatment of dynamics in the ventricular and lumbar CSF extends the new theory and shows that CSF flow rate is the most relevant parameter for understanding the pathological changes of both blood- and brain-derived proteins in CSF. The impact on diagnosis of neuro-degenerative diseases is discussed. PMID- 11498084 TI - A fast chemiluminescent method for H(2)O(2) measurement in exhaled breath condensate. AB - BACKGROUND: Breath condensate can give useful information on volatile compounds produced at alveolar level. Actual concentration of H(2)O(2) in breath condensate is dependent on its production at alveolar level and on the efficacy of the detoxifying systems, catalase, glutathione peroxidase, etc. METHODS: In the present paper, a simple chemiluminescent method for the determination of the H(2)O(2) collected in exhaled breath is shown and data of both smokers and nonsmokers volunteers are presented. RESULTS: The chemiluminescent response is linear up to 100 micromol/l H(2)O(2). The analytical sensitivity is about 0.01 micromol/l. Most of the nonsmokers have a H(2)O(2) content lower than 0.05 micromol/l, while smokers have a content ranging from 0.1 to 0.6 micromol/l. PMID- 11498085 TI - Head-to-head comparison of N-terminal pro-brain natriuretic peptide, brain natriuretic peptide and N-terminal pro-atrial natriuretic peptide in diagnosing left ventricular dysfunction. AB - Brain natriuretic peptide (BNP), NT-proBNP and NT-pro-atrial natriuretic peptide (NT-proANP) were measured in blood samples from 57 patients using immunoassays and immunoradiometric assays to evaluate the usefulness as diagnostic markers for the detection of heart failure. For the detection of impaired left ventricular ejection fraction (LVEF), receiver operating characteristic curves showed that BNP had the best diagnostic performance with an area under curve (AUC) of 0.75+/ 0.06. However, NT-proBNP (AUC: 0.67+/-0.07) and NT-proANP (AUC: 0.69+/-0.08) showed no significant difference to BNP. In a further analysis for the detection of resting LVEF <40%, BNP again was the best marker with an AUC of 0.83+/-0.06. NT-proBNP showed only a slightly smaller AUC (0.79+/-0.07). The AUC for NT-proANP was significantly smaller (0.65+/-0.08) compared to BNP. Additionally, BNP and NT proBNP correlated negatively with the resting LVEF (BNP: -0.472, p<0.001; NT proBNP: -0.306, p=0.026), whereas NT-proANP showed no significant correlation. In summary, BNP was the best marker to detect patients with impaired LVEF compared to NT-proBNP and NT-proANP. However, NT-proBNP showed no significant differences to BNP and it is therefore a new promising alternative marker for the detection of left ventricular dysfunction. PMID- 11498086 TI - Circulating cardiac troponin-T in patients before and after renal transplantation. AB - BACKGROUND: The diagnostic and prognostic use of cardiac troponin T (cTnT) in patients with renal failure has been questioned. Raised serum concentrations of cTnT, with no apparent signs of cardiac damage using conventional methods of detection, have been reported. We aimed to relate circulating concentrations of cTnT to improved renal function following renal transplantation over a one-year period. METHOD: Plasma cTnT was analysed from patients with end stage renal disease before and after transplantation and subsequently at 1, 3, 6 and 12 months. Eight patients had diabetes, 14 had hyperlipidaemia, 8 were smokers and 4 were ex-smokers; all were hypersensitive. RESULTS: At the time of transplantation, 3 of the 32 patients (9.4%) had plasma cTnT concentrations above 0.1 microg/l. In addition to these three patients, five others showed raised cTnT over the one-year period. CONCLUSIONS: The overall trend in circulating cTnT concentrations did not seem to be affected by improved renal function. However, all of the patients that had raised cTnT concentrations at any stage of the one year period had explainable pathologies or were exposed to multiple cardiac risk factors. PMID- 11498087 TI - Multiplex PCR typing of the three most frequent HLA alleles in celiac disease. PMID- 11498090 TI - Living-donor transplants: part of the answer to organ shortage. PMID- 11498091 TI - Current issues and problems of transplantation in the Middle East: the Arabian Gulf. PMID- 11498092 TI - Current issues and future problems in transplantation in East Asia. PMID- 11498093 TI - Comparison of some socioeconomic characteristics of donors and recipients in a controlled living unrelated donor renal transplantation program. PMID- 11498094 TI - Against paid organ donation. PMID- 11498095 TI - Brain death and organ donation in Saudi Arabia. PMID- 11498096 TI - Current issues and future problems of transplantation in the Middle East: Syria. PMID- 11498097 TI - Attitude of ICU staff in Kuwait regarding organ donation and brain death. PMID- 11498098 TI - The psychological effects of kidney donation on living kidney donors (related and unrelated). PMID- 11498099 TI - The complications and morbidity of flank incision for living renal donor. PMID- 11498100 TI - Registries in the Middle East: problems and prospects. PMID- 11498101 TI - Solid organ registry: organization and structure. PMID- 11498102 TI - Renal transplantation from living unrelated donors. PMID- 11498103 TI - Results of 1020 renal transplants single-center experience. PMID- 11498104 TI - Renal transplantation in Iran: a single-center study. PMID- 11498105 TI - Single-centre experience of living related donor nephrectomy. PMID- 11498106 TI - Renal transplantation in Iran. PMID- 11498107 TI - Ten-year experience with related and unrelated living kidney transplantation at one center. PMID- 11498108 TI - Effect of donor nephrectomy on renal function and blood pressure. PMID- 11498109 TI - Reasons for transplant nephrectomy: a retrospective study of 60 cases. PMID- 11498110 TI - Outcome of renal transplantation: 7-year experience. PMID- 11498111 TI - Outcome of living unrelated (commercial) renal transplantation: report of 20 cases. PMID- 11498112 TI - Modified extravesical ureteroneocystostomy and routine ureteral stenting in renal transplantation: experience in 300 consecutive cases. PMID- 11498113 TI - The role of stent and cotrimoxazole in prevention of UTI after kidney transplantation. PMID- 11498114 TI - The efficacy of ureteral stents in prevention of urological complications in renal transplantation. PMID- 11498116 TI - Novel approaches to kidney transplantation. PMID- 11498115 TI - Simultaneous previously failed graft nephrectomy and the use of same vessels for vascular anastomosis in second renal transplantation. PMID- 11498117 TI - Laparoscopic donor nephrectomy: overcoming the learning curve. PMID- 11498118 TI - Laparoscopic nephroureterectomy in a transplant patient with undescended testis. PMID- 11498119 TI - Renal transplantation in patients with abnormal lower urinary tract. PMID- 11498120 TI - Results of a small series of renal graft biopsies in a Lebanese population. PMID- 11498121 TI - Early and late effects of renal transplantation on cardiac functions. PMID- 11498122 TI - Postoperative surgical complications in renal transplant recipients: one center's experience. PMID- 11498123 TI - Vascular complications in renal transplantation. PMID- 11498124 TI - Cardiac response to stimulation of baroreflex of carotid artery in renal transplant patients. PMID- 11498125 TI - Evaluation of serum levels of lipids and lipoproteins in kidney-transplanted patients. PMID- 11498126 TI - Chronic renal allograft dysfunction: immunologic and nonimmunologic risk factors. PMID- 11498127 TI - Chronic rejection of renal grafts: the role of acute rejection. PMID- 11498128 TI - Chronic graft dysfunction: donor factors. PMID- 11498129 TI - Does recipient age affect the outcome of renal transplantation? PMID- 11498130 TI - Hyperhomocysteinemia in renal transplant recipients. PMID- 11498131 TI - Alport's syndrome and renal transplantation. PMID- 11498132 TI - Donor referral pattern for liver transplantation in Saudi Arabia. PMID- 11498133 TI - The first 100 liver transplants in Saudi Arabia. PMID- 11498134 TI - Vascular complications in the first 100 liver transplantations in Saudi Arabia. PMID- 11498135 TI - Outcome and risk factors of hepatic artery thrombosis after orthotopic liver transplantation in adults. PMID- 11498136 TI - Successful strategy for reducing biliary complications after liver transplantation. PMID- 11498137 TI - Recurrent disease after liver transplantation. PMID- 11498138 TI - Factors involved in the functional recovery of a liver graft with particular emphasis on liver steatosis: a multivariate analysis. PMID- 11498139 TI - Liver transplantation in patients with previous portasystemic shunt. PMID- 11498140 TI - Living-donor split-liver transplantation. PMID- 11498142 TI - Imaging findings and interventional treatments for complications in partial-liver transplant recipients. PMID- 11498141 TI - Donor safety in living-related liver transplantation. PMID- 11498143 TI - Long-term efficacy of endoscopic stenting in patients with stricture of the biliary anastomosis after orthotopic liver transplantation. PMID- 11498144 TI - Liver transplantation in an adolescent with Alagille syndrome. PMID- 11498146 TI - Living-related donor transplantation other than kidney. PMID- 11498145 TI - Liver transplantation for ecstasy-induced fulminant hepatic failure. PMID- 11498147 TI - Molecular and cellular mechanisms of donor heart dysfunction. PMID- 11498148 TI - Effect of ABO blood type matching in cardiac transplant recipients. PMID- 11498149 TI - Morphologic changes in heterotopically transplanted rat heart isografts. PMID- 11498150 TI - Etiology of pulmonary nodules after heart transplantation. PMID- 11498151 TI - Heart retransplantation: institutional results of a series of 31 cases. PMID- 11498152 TI - A novel combination therapy to reverse end-stage heart failure. PMID- 11498153 TI - Induction immunosuppression with interleukin-2 receptor antibodies (basiliximab and daclizumab) in renal transplant recipients. PMID- 11498154 TI - Successful therapy of acute vascular rejection with combined plasma-exchange and monoclonal antibody. PMID- 11498155 TI - The comparison of angiotensin-converting enzyme inhibitors with calcium antagonists in renal transplant patients under cyclosporin treatment. PMID- 11498156 TI - Improved primary transplant success rates using a triple regimen of cyclosporine microemulsion, mycophenolate mofetil and prednisone. PMID- 11498157 TI - Therapeutic drug level monitoring for mycophenolate mofetil: is it justified? PMID- 11498158 TI - The cost effectiveness of mycophenolate mofetil in the first year after living related renal transplantation. PMID- 11498159 TI - Cyclosporine lymphocyte versus whole blood pharmacokinetic monitoring: correlation with histological findings. PMID- 11498160 TI - High dose steroid regime: a tool for reducing organ rejection. PMID- 11498161 TI - Donor specific transfusion in kidney transplantation: effect of different immunosuppressive protocols on graft outcome. PMID- 11498162 TI - High incidence of proteinuria in hepatitis C virus-infected renal transplant recipients is associated with poor patient and graft outcome. PMID- 11498163 TI - Ferritin levels can be a good predictor for HCV viremia in potential renal transplant candidates. PMID- 11498164 TI - Cytomegalovirus infection in kidney transplant recipients: early diagnosis and monitoring of antiviral therapy by the antigenemia assay. PMID- 11498165 TI - Is there risk of transmitting hepatitis B virus in accepting hepatitis B core antibody-positive donors for living related liver transplantation? PMID- 11498166 TI - Prevalence of tuberculosis after renal transplantation in Iran. PMID- 11498167 TI - Incidence and clinical characteristics of malignancies after renal transplantation: one center's experience. PMID- 11498168 TI - Incidence of postrenal transplantation malignancies: a report of two centers in Tehran, Iran. PMID- 11498169 TI - Cancer after renal transplantation in Kuwait. PMID- 11498170 TI - Posttransplant lymphoproliferative disorders: report from a single center. PMID- 11498171 TI - Uterine cervical intraepithelial neoplasia in renal transplantation. PMID- 11498172 TI - Kidney transplantation and Kaposi's sarcoma: review of 2050 recipients. PMID- 11498173 TI - Intensive care and immediate follow-up of children after renal transplantation. PMID- 11498174 TI - Renal transplantation in children: results of 9 years of renal transplantation in children at our center. PMID- 11498175 TI - Renal transplantation in children. PMID- 11498176 TI - Impact of donor nephron mass on outcomes in renal transplantation. PMID- 11498177 TI - Renal tubular dysfunction following kidney transplantation: a prospective study in 31 children. PMID- 11498178 TI - Heterotopic liver transplantation during adolescence. PMID- 11498179 TI - Outcome of pediatric heart transplantation: an analysis of 27 cases. PMID- 11498180 TI - Major histocompatibility complex class I and II antigens frequencies in selected groups of Lebanese. PMID- 11498181 TI - Transplant outcome in patients with non-O blood groups who received kidneys from O blood group donors. PMID- 11498182 TI - The effect of simvastatin on panel-reactive antibody and crossmatch positivity. PMID- 11498183 TI - Panel-reactive antibodies using complement-dependent cytotoxicity, flow cytometry, and ELISA in patients awaiting renal transplantation or transplanted patients: a comparative study. PMID- 11498184 TI - Crossmatch in renal transplantation using a sensitive antiglobulin microlymphocytotoxicity test. PMID- 11498185 TI - Soluble HLA class I molecules as immune regulators in transplantation. PMID- 11498186 TI - Peripheral blood microchimerism in female renal recipients from male donors. PMID- 11498187 TI - Evaluation of the effect of fish oil on cell kinetics: implications for clinical immunosuppression. PMID- 11498188 TI - HBV and HCV infection before and after kidney transplantation. PMID- 11498189 TI - Scintigraphic evaluation of graft and native liver function after heterotopic liver transplantation. PMID- 11498190 TI - Aberrant T-cell activation via CD95 and apoptosis in peripheral T lymphocytes in stable heart transplant recipients. PMID- 11498191 TI - Marrow transplantation in advanced thalassemic patients using a new conditioning regimen. PMID- 11498192 TI - Gene therapy of ischemic-damaged kidney in the rat using hepatocyte growth factor gene. PMID- 11498193 TI - Ischemia/reperfusion injury of rat small intestine: the effect of allopurinol dosage. PMID- 11498194 TI - Long-term follow-up of liver transplant recipients undergoing cyclosporine withdrawal. PMID- 11498195 TI - Return to work 5 years after liver transplantation. PMID- 11498197 TI - Diabetes mellitus in transplantation. Introduction. PMID- 11498198 TI - Diabetes mellitus: a major challenge in transplantation. PMID- 11498199 TI - Risk factors and incidence of posttransplant diabetes mellitus. PMID- 11498200 TI - Clinical impact of posttransplant diabetes mellitus. PMID- 11498201 TI - Impact of immunosuppressive regimes on posttransplant diabetes mellitus. PMID- 11498202 TI - Strategies for risk reduction and management of posttransplant diabetes mellitus. PMID- 11498203 TI - Clinical impact and health economic consequences of posttransplant type 2 diabetes mellitus. PMID- 11498204 TI - Will the good doctors please stand up? PMID- 11498205 TI - Comparative symptomatology of West Nile fever. PMID- 11498206 TI - Prospects for prevention of meningococcal meningitis. PMID- 11498207 TI - A role for phosphodiesterase type-4 inhibitors in COPD? PMID- 11498209 TI - Intestinal permeability and morphometric recovery in coeliac disease. PMID- 11498208 TI - Osteoprotegerin: a link between osteoporosis and arterial calcification? PMID- 11498210 TI - Pricing and reimbursement of pharmaceuticals in the Baltic States. PMID- 11498211 TI - Epidemic West Nile encephalitis, New York, 1999: results of a household-based seroepidemiological survey. AB - BACKGROUND: In the summer of 1999, West Nile virus was recognised in the western hemisphere for the first time when it caused an epidemic of encephalitis and meningitis in the metropolitan area of New York City, NY, USA. Intensive hospital based surveillance identified 59 cases, including seven deaths in the region. We did a household-based seroepidemiological survey to assess more clearly the public-health impact of the epidemic, its range of illness, and risk factors associated with infection. METHODS: We used cluster sampling to select a representative sample of households in an area of about 7.3 km(2) at the outbreak epicentre. All individuals aged 5 years or older were eligible for interviews and phlebotomy. Serum samples were tested for IgM and IgG antibodies specific for West Nile virus. FINDINGS: 677 individuals from 459 households participated. 19 were seropositive (weighted seroprevalence 2.6% [95% CI 1.2-4.1). Six (32%) of the seropositive individuals reported a recent febrile illness compared with 70 of 648 (11%) seronegative participants (difference 21% [0-47]). A febrile syndrome with fatigue, headache, myalgia, and arthralgia was highly associated with seropositivity (prevalence ratio 7.4 [1.5-36.6]). By extrapolation from the 59 diagnosed meningoencephalitis cases, we conservatively estimated that the New York outbreak consisted of 8200 (range 3500-13000) West Nile viral infections, including about 1700 febrile infections. INTERPRETATION: During the 1999 West Nile virus outbreak, thousands of symptomless and symptomatic West Nile viral infections probably occurred, with fewer than 1% resulting in severe neurological disease. PMID- 11498213 TI - Pregnancy outcomes after peripheral blood or bone marrow transplantation: a retrospective survey. AB - BACKGROUND: Some patients treated by transplantation of haemopoietic stem cells (peripheral blood or bone marrow) become permanently infertile, but others retain or recover fertility. We assessed the outcome of conception in women, and partners of men previously treated by autologous or allogeneic stem cell transplantation (SCT). METHODS: We sent questionnaires to 229 centres of the European Group for Blood and Marrow Transplantation. We sought details about the original disease, transplant procedure, and outcome of conception for both male and female patients. FINDINGS: 199 centres gave information relating to 19412 allogeneic and 17950 autologous transplant patients. 232 (0.6%) patients conceived after SCT. Crude annual birth rate for 4-month survivors of SCT was lower than the national average for England and Wales at 1.7 per 1000 patients. 312 conceptions were reported in 113 patients (74 allograft) and partners of 119 patients (93 allograft). Most pregnancies were uncomplicated and resulted in 271 livebirths. 28 (42%) of 67 allograft recipients had caesarean section compared with 16% in the normal population (difference =26% [95% CI 15-38]), 12 (20%) of 59 had preterm delivery compared with a normal rate of 6% (14% [4-24]), and 12 (23%) of 52 had low birthweight singleton offspring compared with a normal rate of 6% (17% [6-29]). INTERPRETATION: Pregnancy after SCT is likely to have a successful outcome. Pregnancies in allograft patients who have received total body irradiation should be treated as high risk for maternal and fetal complications. PMID- 11498212 TI - Cilomilast, a selective phosphodiesterase-4 inhibitor for treatment of patients with chronic obstructive pulmonary disease: a randomised, dose-ranging study. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a common, progressive respiratory disease that causes great morbidity and mortality despite treatment. There is evidence for airway inflammation in COPD. Cilomilast is an orally active, potent, selective phosphodiesterase type 4 inhibitor, which in vitro can affect cells thought to be of clinical importance in COPD. Our aim was to assess the safety, efficacy, and dose response of cilomilast in the treatment of patients with this disease. METHODS: We did a 6-week, randomised, dose-ranging study in 424 patients with COPD (forced expiratory volume in 1 s [FEV(1)] 46.8% of predicted, FEV(1)/forced vital capacity [FVC] 54.6%, and postsalbutamol reversibility 5.4%). We randomly assigned individuals at 60 European centres to receive cilomilast 5 (n=109), 10 (n=102), or 15 (n=107) mg twice daily, or placebo (n=106). The main outcome measure was trough FEV(1) before and after use of a bronchodilator. Analyses were by intention to treat. FINDINGS: Cilomilast 15 mg twice daily significantly improved FEV(1) compared with placebo (mean 130 mL vs -30 mL [95% CI 90-240] at week 6, p<0.0001). FVC and peak expiratory flow were also improved (p=0.001 and p<0.0001, respectively). Quality of life measures did not differ significantly between the groups. There were no significant differences in serious adverse events between the groups. INTERPRETATION: Cilomilast 15 mg twice daily might be an effective maintenance treatment for COPD. Further clinical studies are underway. PMID- 11498214 TI - Polychemotherapy for early breast cancer: an overview of the randomised clinical trials with quality-adjusted survival analysis. AB - BACKGROUND: Overview analysis involving 18000 women with breast cancer in 47 randomised trials showed that prolonged chemotherapy significantly reduces the risk of relapse and death compared with no chemotherapy. Here we express the size of the benefit in terms of quality-adjusted survival time gained. METHODS: We used the Q-TWiST method (Quality-adjusted Time Without Symptoms of disease and Toxicity of treatment) to provide treatment comparisons within 10 years' follow up, incorporating differences in quality of life associated with times patients spend with chemotherapy toxic effects, after relapse, and without symptoms of relapse or toxicity. FINDINGS: Within 10 years' follow-up the benefit of increased relapse-free and overall survival for younger women (<50 years old) who received polychemotherapy balanced the burdens in terms of acute toxic side effects, especially among women enrolled in trials that did not include tamoxifen. Overall, chemotherapy-treated younger women gained an average of 10.3 months of relapse-free survival and 5.4 months of overall survival within 10 years (p<0.0001 for both) compared with the no-chemotherapy group. Polychemotherapy provided more quality-adjusted time than control across nearly all values of utility weights for time spent undergoing chemotherapy and time after relapse. The range of benefit was from -0.6 to 10.3 months. For older women (50-69 years) overall, polychemotherapy also provided significant benefit compared with no chemotherapy but, compared with younger women, the size of benefit was less and the range of utility-weight values favouring polychemotherapy was smaller. Average gains for older women treated with polychemotherapy (with or without tamoxifen) were 6.8 months of relapse-free survival (p<0.0001) and 2.9 months of overall survival (p=0.0001) within 10 years. The range of quality-adjusted benefit was -3.1 to 6.8 months. For older women with oestrogen-receptor-poor tumours who did not receive tamoxifen (9% of the total), the benefit of polychemotherapy was significant and similar to that observed for younger women. INTERPRETATION: The benefits of adjuvant chemotherapy within 10 years outweigh the burdens especially for younger women (<50 years old) and among older women (50-69 years) to a lesser degree. Additional studies to compare the quality-adjusted survival of chemotherapy plus endocrine therapy versus endocrine therapy alone are required for younger patients with tumours that express steroid-hormone receptors. PMID- 11498215 TI - Timely detection of meningococcal meningitis epidemics in Africa. AB - BACKGROUND: Epidemics of meningococcal disease in Africa are commonly detected too late to prevent many cases. We assessed weekly meningitis incidence as a tool to detect epidemics in time to implement mass vaccination. METHODS: Meningitis incidence for 41 subdistricts in Mali was determined from cases recorded in health centres (1989-98) and from surveillance data (1996-98). For incidence thresholds of 5 to 20 cases per 100000 inhabitants per week, we calculated sensitivity and specificity for detecting epidemics, and determined the time lapse between threshold and epidemic peak. FINDINGS: We recorded 9084 meningitis cases. Clinic-based weekly incidence of 5 and 10 cases per 100000 inhabitants detected all meningitis epidemics (sensitivity 100%, 95% CI 93-100), with median threshold-to-peak time of 5 and 3 weeks. Under-reporting reduced sensitivity: only surveillance thresholds of 5 or 7 cases per 100000 inhabitants per week detected all epidemics. Crossing the lower threshold before the 10th calendar week doubled epidemic risk relative to crossing it later (relative risk 2.1, 95% CI 1.4-3.2). At 10 cases per 100000 inhabitants per week, specificity for outbreak prediction was 88%, 95% CI 83-91). For populations under 30000, 3 to 5 cases in one or two weeks predicted epidemics with 85% to 97% specificity. INTERPRETATION: Low meningitis thresholds improve timely detection of epidemics. Ten cases per 100000 inhabitants per week in one area confirm epidemic activity in a region, with few false alarms. An alert threshold of 5 cases per 100000 inhabitants per week allows time to investigate, prepare for an epidemic, and initiate mass vaccination where appropriate. For populations under 30000, the alert threshold is two cases in a week. High quality surveillance is essential. PMID- 11498216 TI - Clinical picture: Oesophageal varix bleed. PMID- 11498217 TI - An unusual case of sarcoidosis. PMID- 11498218 TI - Effect of infliximab on sight-threatening panuveitis in Behcet's disease. AB - Permanent loss of vision resulting from relapsing ocular inflammation occurs frequently in patients with Behcet's disease, despite intensive, chronic immunosuppressive therapy. Since tumour necrosis factor (TNF) might have an important pathogenetic role in Behcet's disease, we decided to give a single infusion of infliximab-a monoclonal antibody against TNF-to five patients with relapsing panuveitis, at the immediate onset of last relapse. Remission of ocular inflammation was evident within the first 24 h, and complete suppression was seen 7 days after treatment in all patients. No side-effects were noted. We suggest that infliximab is a rapid and effective new therapy for sight-threatening ocular inflammation in Behcet's disease. PMID- 11498219 TI - Incidence of Hodgkin's disease in Nordic countries. AB - We analysed age and sex-specific trends in the incidence of Hodgkin's lymphoma in Denmark, Sweden, Norway, and Finland during 1978-97. The incidence of Hodgkin's lymphoma decreased significantly in all age groups above 40 years; there was a significant increase in incidence among adolescents and young adults. This increase occurred primarily for Hodgkin's lymphoma of the nodular sclerosis subtype. These observations emphasise the need to identify risk factors for Hodgkin's lymphoma in the young. PMID- 11498220 TI - OLIG2 as a specific marker of oligodendroglial tumour cells. AB - OLIG2 is a recently identified transcription factor involved in the specification of cells in the oligodendroglial lineage. We investigated the expression of OLIG2 by in-situ hybridisation in 21 brain tumours: nine grade II and III oligodendrogliomas, three grade II oligoastrocytomas, and nine non oligodendroglial tumours (four grade IV astrocytomas, two meningiomas, a dysembryoplastic neuroepithelial tumour, and two metastases). OLIG2-positive cells corresponding to neoplastic oligodendrocytes were present in all oligodendrogliomas and oligoastrocytomas. By contrast, OLIG2 expression was not detected in the non-oligodendroglial tumours. Thus, oligodendroglioma probably arise from oligodendrocyte precursor cells. OLIG2 should prove a useful marker for the diagnosis of oligodendroglial tumours. PMID- 11498221 TI - Public health in Afghanistan plunges to new depths. PMID- 11498222 TI - New genes reveal major role for iron in neurodegeneration. PMID- 11498223 TI - Small part of cannabis puzzle solved by animal studies. PMID- 11498224 TI - Link between gum disease and heart disease disputed. PMID- 11498225 TI - Researchers identify two populations of neural stem cells. PMID- 11498226 TI - New variant CJD fears threaten blood supplies. PMID- 11498227 TI - Japan reviews policy on mental illness and crime. PMID- 11498228 TI - European Commission may reform drug advertising legislation. PMID- 11498231 TI - Israel Medical Association proposes "rescue project" for health. PMID- 11498232 TI - Kenya government promises to increase doctors' salaries to curb brain drain. PMID- 11498233 TI - Information technology can benefit developing countries. PMID- 11498234 TI - Turner's syndrome. AB - Before chromosomal analysis became available, the diagnosis of Turner's syndrome was based on the characteristics independently described by Otto Ullrich and Henry Turner, such as short stature, gonadal dysgenesis, typical, visible dysmorphic stigmata, and abnormalities in organs, which present in individuals with a female phenotype. Today, Turner's syndrome or Ullrich-Turner's syndrome may be defined as the combination of characteristic physical features and complete or part absence of one of the X chromosomes, frequently accompanied by cell-line mosaicism. The increasing interest in Turner's syndrome over the past two decades has been motivated both by the quest for a model by which the multi faceted features of this disorder can be understood, and the endeavour to provide life-long support to the patient. New developments in research allow patients with Turner's syndrome to have multidisciplinary care. PMID- 11498235 TI - Social medicine in Latin America: productivity and dangers facing the major national groups. AB - There is little knowledge about Latin American social medicine in the English speaking world. Social medicine groups exist in Argentina, Brazil, Chile, Colombia, Cuba, Ecuador, and Mexico. Dictatorships have created political and economic conditions which are more adverse in some countries than others; in certain instances, practitioners of social medicine have faced unemployment, arrest, torture, exile, and death. Social medicine groups have focused on the social determinants of illness and early death, the effects of social policies such as privatisation and public sector cutbacks, occupational and environmental causes of illness, critical epidemiology, mental health effects of political trauma, the impact of gender, and collaborations with local communities, labour organisations, and indigenous people. The groups' achievements and financial survival have varied, depending partly on the national context. Active professional associations have developed, both nationally and internationally. Several groups have achieved publication in journals and books, despite financial and technical difficulties that might be lessened through a new initiative sponsored by the US National Library of Medicine. The conceptual orientation and research efforts of these groups have tended to challenge current relations of economic and political power. Despite its dangers, Latin American social medicine has emerged as a productive field of work, whose findings have become pertinent throughout the world. PMID- 11498241 TI - Lost in space. PMID- 11498237 TI - Gaucher's disease. PMID- 11498242 TI - The production and bioactivity of rainbow trout (Oncorhynchus mykiss) recombinant IL-1 beta. AB - The predicted rainbow trout mature interleukin-1 beta (IL-1 beta) peptide has been produced as a recombinant protein in E. coli. The bioactivity of this molecule has been studied using trout head kidney cell preparations and a trout macrophage cell line (RTS11). Trout rIL-1 beta was shown to increase the expression level of IL-1 beta, cyclooxygenase (COX2) and MHC class II beta chain transcription, as determined by Northern blot analysis. Stimulatory doses of rIL 1 beta were typically > or =10 ng/ml. Induction of IL-1 beta expression occurred within 1h post-stimulation with trout rIL-1 beta and was maximal 3-6h post stimulation. Trout rIL-1 beta was also able to increase murine D10.G4.1 cell proliferation and trout head kidney leukocyte phagocytic activity, in a dose dependent manner. However, equivalent D10.G4.1 cell proliferation was induced with approximately 1000-fold lower doses of human rIL-1 beta. That LPS contamination did not contribute to the effects seen was confirmed by determining its concentration in the trout rIL-1 beta preparation, and demonstrating that the rIL-1 beta activity was inhibited by heating or pre-incubation with a polyclonal anti-trout rIL-1 beta antibody. PMID- 11498243 TI - Novel transcripts encoding secreted forms of feline CD80 and CD86 costimulatory molecules. AB - Engagement of costimulatory molecules such as CD28 or CD152 (CTLA4) on T cells by CD80 (B7-1) or CD86 (B7-2) dictates the nature of T cell-mediated immune responses. We previously reported the discovery of naturally occurring forms of canine CD80 and CD86 mRNAs which encode secreted CD80 and CD86 molecules. We report here that mRNAs for secreted forms of CD80 and CD86 are also expressed in cats. The mRNA for secreted feline CD86 is generated by deleting a transmembrane domain exon, which is the same mechanism we described for secreted canine CD86. We also identified a feline CD80 transcript that only retains the immunoglobulin variable-like domain. The detection of naturally occurring mRNAs encoding secreted CD80 and CD86 adds further complexity to the regulation of the B7 CD28/CD152 costimulatory pathway. PMID- 11498244 TI - Cyclosporin immunosuppression of sheep: pharmacokinetics and allograft survival. AB - A chronically immunosuppressed sheep model was established using a regimen of cyclosporin A (CsA; 2-3mg/kg twice daily) and ketoconazole (10mg/kg twice daily). Blood CsA concentrations reached a steady-state after 17 days of treatment. The clearance of CsA decreased from a mean (95% CI) of 9.47 (6.2-12.7)ml/min/kg after a single (first) dose (3mg/kg i.v.) to 1.62 (1.38-1.86)ml/min/kg after 18 days of CsA (3mg/kg i.v. twice daily) co-administration with ketoconazole. These data indicated that the combination of CsA and ketoconazole could be used to give stable high concentrations of CsA in the sheep. Using this regimen in the sheep, the long-term survival of skin allografts was monitored as an indicator of effective immunosuppression. CsA in blood was measured daily and CsA dose adjusted to various target concentration ranges. Provided that the trough concentration of blood CsA was maintained between 1500-2500 mg/l, long-term healthy skin allografts were maintained on the sheep without significant adverse effects on haematological or biochemical parameters. PMID- 11498245 TI - Changes in peripheral blood leukocyte populations in pigs with natural postweaning multisystemic wasting syndrome (PMWS). AB - The objective of the present study was to analyze, by flow cytometry, changes in PBMC subsets in pigs having postweaning multisystemic wasting syndrome (PMWS), a new condition associated to porcine circovirus type 2 (PCV2) infection. Thirteen acutely PMWS affected pigs were selected from a farm seronegative to porcine reproductive and respiratory syndrome virus (PRRSV) and to Aujeszky's disease virus (ADV); 11 clinically healthy pigs were selected from a high health farm with no history of PMWS and free of the major swine pathogens, and used as a control group. All pigs were necropsied, and tissue samples were fixed in formalin; blood with EDTA anticoagulant was used to perform the flow cytometric analysis. PBMC were incubated with mAb against porcine CD3, CD4, CD8, CD25, CD45, IgM, SWC3, and SLA-Class II. Flow cytometric analysis showed substantial changes in leukocyte subsets in the peripheral blood of PMWS-affected pigs, which were characterized by an increase of monocytes, a reduction of T (mainly CD4(+)) and B lymphocytes, and the presence of low-density immature granulocytes. Altogether, these changes would suggest an inability of acutely PMWS-affected pigs to mount an effective immune response. PMID- 11498246 TI - Expression, purification, and in vitro biological activities of recombinant bovine granulocyte-colony stimulating factor. AB - Neutrophils are essential components of the innate immune system and they play a critical role in the defense of host against bacterial and fungal infections. The colony stimulating factors are a class of glycoproteins that are required for proliferation, differentiation, and functional activation of hematopoietic progenitor cells. Granulocyte-colony stimulating factor (G-CSF) is a member of this regulatory family of cytokines that specifically stimulates proliferation and maturation of precursor cells in the bone marrow into fully differentiated and functional neutrophils. G-CSF also modulates the biological activities of mature neutrophils in circulation. A bovine G-CSF (bG-CSF) cDNA clone (previously isolated and sequenced in our laboratory) was expressed in Escherichia coli and the biological activities of the solubilized protein from purified inclusion bodies were examined. Flow cytometric analysis of membrane antigen density of neutrophils activated with bG-CSF revealed an upregulation in the expression of CD11a (>114%), CD11b (>148%), CD11c (>87%), and CD18 (>109%). Expression of L selectin was decreased by more than 43%. There was no change, however, in the expression of CD14. These findings indicate that recombinant bG-CSF (rbG-CSF) expressed in E. coli is biologically active and exerts the same type of effects on neutrophils in vitro as those of human G-CSF (hG-CSF). PMID- 11498247 TI - Oral administration of IL-1 beta enhanced the proliferation of lymphocytes and the O(2)(-) production of neutrophil in newborn calf. AB - Recently, we demonstrated the presence of IL-1 beta in the colostral whey from dairy cows. Here, authors examined oral transmission of colostral IL-1 beta and its immunological effects on the neonatal calves. Biotin-labeled recombinant bovine (rb) IL-1 beta was administered orally to newborn calves and monitored in the serum. The results disclosed the passive transfer of colostral cytokines via the oral route, and a potent increase in white blood cell (WBC) count was observed in all calves administered with rbIL-1 beta. Oral administration of IL-1 beta significantly increased the proliferation of peripheral blood mononuclear cells (PBMCs) stimulated with concanavalin A, and the O(2)(-) production of stimulates neutrophils in newborn calves. These results suggest that the oral administration of IL-1 beta has an immunostimulatory activity in the newborn calf. PMID- 11498248 TI - Induction of inflammatory cytokines by extracellular products and LPS of the fish pathogen Aeromonas salmonicida ssp. achromogenes in mice and mouse cell cultures. AB - Balb/c mice, injected i.p. with extracellular products (ECP) of Aeromonas salmonicida subsp. achromogenes (Asa), displayed symptoms similar to toxic shock syndrome. The LD(50) observed was between 1.5 and 2.0 microg g(-1) and the mice died within 19 h. Four inflammatory cytokines were measured in mice receiving sublethal ECP doses. TNF-alpha and IL-6 showed a sharp peak in the serum while IL 1 beta and IL-2 were not detected. When peritoneal macrophages were cultivated in the presence of ECP, AsaP1 (a toxic caseinolytic metallo-protease purified from ECP) or LPS, all cultures produced TNF-alpha, IL-6 and IL-1 beta. The same antigens were mitogenic in spleen cell cultures. Furthermore, IL-2 production, which is a normal T-cell response to ConA stimulation, was downregulated in spleen cell cultures from mice injected with ECP. PMID- 11498249 TI - IFN gamma and IL-4 production by CD4, CD8 and WC1 gamma delta TCR(+) T cells from cattle lymph nodes and blood. AB - The synthesis of IFN gamma and IL-4 by CD4, CD8 and WC1 gamma delta TCR(+) T cell sub-populations, and T cells stained with activation/memory-sub-set markers has been examined by flow cytometric analysis. Cells from blood, prescapular, bronchial and mesenteric lymph nodes and Peyer's patches were incubated with phorbol 12-myristate 13-acetate (PMA), ionomycin and brefeldin-A before staining. Lymphocytes that stained for cytoplasmic IFN gamma were evident within the CD4 and CD8 populations from all tissues and also in the WC1 population from lymph nodes. IL-4 producing cells were primarily evident within the CD4 population. IFN gamma synthesis was evident within both CD45RO(+) and CD45RB(+) populations, but IL-4 synthesis was predominantly by cells that were CD45RO(+)/CD45RB(-). Expression of CD62L is not related to functional memory in CD4(+) T cells from cattle and CD62L(+) cells, particularly from the lymph nodes draining the skin and the lungs, stained with mAb to IFN gamma and IL-4. The findings indicate that at least for CD4(+) T cells, where CD45 isoform expression is related to functional memory, these two cytokines are produced predominantly by cells with a memory phenotype. The observation that some WC1(+) cells produce IFN gamma implies the presence of distinct sub-sets of this gamma delta TCR(+) population cattle and suggests a functional role. PMID- 11498250 TI - Comparative analysis of porcine cytokine production by mRNA and protein detection. AB - To analyze the correlation between cytokine mRNA transcription and secretion, interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) were quantified by enzyme linked immunosorbent assay and IL-2 by bioassay and compared with their mRNA levels, determined by reverse transcription polymerase chain reaction (RT-PCR). For this purpose, peripheral blood monomorphonuclear cells (PBMC) were stimulated in vitro with the lectins pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (ConA), respectively, and with F4 fimbriae in an antigen-specific assay. Analyses were performed 4, 8, 16, 24 and 48 h after stimulation on the stimulated PBMC for mRNA and on the respective culture supernatants for proteins. RT-PCR products were quantified by densitometric scanning of the electrophoresis bands and related to the band intensity of the housekeeping gene, cyclophilin. Low levels of IL-2, IL-4 and IFN-gamma mRNA were detected in unstimulated PBMC. Stimulation with all three mitogens (PWM, ConA, PHA) led to an increase in mRNA transcription. In contrast, substantial IL-10 mRNA levels were detected in both unstimulated and stimulated cells with practically no difference between the three mitogens used. IL-2 mRNA expression tended to peak after 8-16 h for all three mitogens. The cells stimulated with PWM and ConA showed higher levels of gene expression for IFN-gamma and lower for IL-4 then the cells stimulated with PHA, however, differences were not statistically significant. For cells stimulated with F4 fimbriae only the IFN-gamma mRNA expression increased with an early peak at 8h post-stimulation. The analysis of the culture supernatants for secreted cytokines revealed a correlation between the levels of mRNA transcription and the respective secreted cytokines during the first 24h of stimulation. After 24h of stimulation, however, a decrease in IFN-gamma and IL-2 mRNA levels was accompanied by an increase or a less pronounced decrease in cytokine concentration; only the ConA induced IL-2 mRNA and protein concentration slopes showed similar profiles. In conclusion, similar cytokine production profiles as defined by mRNA and protein, respectively are obtained only during the first 24h after stimulation of the cell cultures. PMID- 11498251 TI - Ontogeny of channel catfish lymphoid organs. AB - Previously, we showed that catfish could not mount a detectable antibody response after bacterial exposure until 21 days post-hatch (ph). In order to evaluate the changes associated with the development of a functional humoral response, we evaluated the temporal and spatial distribution of immune cell populations in developing catfish. Cells functioning in nonspecific immunity were present in the renal hematopoietic tissue (rht) and thymus at hatch and in the spleen by day 3 ph. Immunoglobulin (Ig) positive lymphocytes were first detected on day 7, 10, and 14 in the rht, thymus and spleen, respectively. Mature thymocytes were first detected on day 10 ph. Distinct thymic regionalization and splenic lymphoid tissue organization were not observed until day 21 ph. We suggest that the reason for a lack of antibody production until day 21 ph is the poor organization of secondary lymphoid tissue until that age. PMID- 11498252 TI - Tumor necrosis factor alpha and its receptors in experimentally bovine leukemia virus-infected sheep. AB - To examine whether tumor necrosis factor alpha (TNF alpha) contributes to the pathogenesis of bovine leukemia virus (BLV) infection, the mRNA expression patterns of TNF alpha and its receptors, type 1 (TNF R1) and type 2 (TNF R2) were investigated. Sheep inoculated with BLV were divided into two groups; one was BLV positive and the other BLV-negative based on the detection in peripheral blood mononuclear cells (PBMC). Expression of TNF R1 mRNA was down-regulated in PBMC from the BLV-positive compared to BLV-negative sheep. No difference was shown in the expression levels of TNF R2 mRNA between the two groups. Furthermore, proliferative responses of PBMC in the presence of TNF alpha were observed from the BLV-positive, but not BLV-negative sheep. Membrane-bound TNF alpha (mTNF alpha) is thought to be one of the ligands, inducing B-cell activation. Flow cytometric analysis demonstrated that the number of PBMC, that were positive for mTNF alpha expression, was increased in the BLV-positive sheep. Thus, the expression of TNF alpha and its receptors may be closely associated with lymphocytosis induced by BLV. PMID- 11498253 TI - Fc receptors for immunoglobulin G--a role in the pathogenesis of Guillain-Barre syndrome and multiple sclerosis. AB - Receptors for the Fc part of IgG (FcgammaR) constitute a family of cell-surface molecules expressed on almost every cell of the immune system. They are also present on non-lymphoid cells such as Schwann cells and endothelial cells. By linking humoral and cell-mediated responses, FcgammaR are key in defending against pathogens. Polymorphisms in some FcgammaR genes are associated with infectious and autoimmune diseases. It was found recently that certain FcgammaRIIA and FcgammaRIIIB allotypes are correlated with the disease course of Guillain-Barre syndrome (GBS) and multiple sclerosis (MS). This may imply that clearance of circulating autoantibodies and immune complexes is important in the pathogenesis of these diseases. PMID- 11498254 TI - In vitro evidence that immunuaffinity-purified MOG contains immunogenic quantities of contaminating mouse IgG; techniques for producing Ig-free MOG. AB - Myelin oligodendrocyte glycoprotein (MOG) has attracted considerable attention due to its possible role in multiple sclerosis (MS). Previous studies have shown that MOG-reactive T cells are more commonly detected in MS patients than controls. In this report, we studied human MOG-reactive T cell clones generated from healthy individuals as well as MS patients. Our results indicate that immunoaffinity-purified MOG, which was routinely used in prior studies, is contaminated by anti-MOG antibody (mouse IgG), despite the inability to detect IgG by Western blotting. Here, we used continuous SDS-PAGE, which enabled us to isolate highly purified MOG devoid of contaminating mouse IgG. PMID- 11498255 TI - Expression of interleukin-1 beta in rat dorsal root ganglia. AB - The expression of interleukin-1beta was examined in dorsal root ganglion (DRG) neurons from adult rats using non-radioactive in situ hybridization and immunocytochemistry. At all spinal levels, approximately 70% of the DRG neurons appeared to express IL-1beta mRNA; about 80% of these DRG neurons actually appeared to produce the IL-1beta protein at markedly varying levels. The expression of IL-1beta was found in large as well as in intermediate diameter sensory neurons but only sporadically in the population of small sensory neurons. The population of IL-1beta immunopositive sensory neurons included most of the large calretinin-positive Ia afferents, but only a few of the small substance P/CGRP positive sensory neurons. In situ hybridization staining for the detection of type 1 IL-1 receptor showed expression of this receptor by most of the sensory neurons as well as by supportive glial-like cells, presumably satellite cells. The functional significance of IL-1beta in the DRG neurons needs to be elucidated, but we speculate that IL-1beta produced by DRG neurons may be an auto/paracrine signalling molecule in sensory transmission. PMID- 11498256 TI - Experimental autoimmune uveoretinitis in mice (Biozzi ABH and NOD) expressing the autoimmune-associated H-2A(g7) molecule: identification of a uveitogenic epitope. AB - To determine whether Biozzi ABH (H-2A(g7)) mice were susceptible to chronic experimental autoimmune uveoretinitis (EAU). Biozzi ABH were immunized with the two retinal antigens, interphotoreceptor retinoid binding protein (IRBP) and soluble antigen (S-Ag). Biozzi ABH mice were found to be susceptible to EAU induction with native bovine IRBP. Recombinant protein domains were used to identify IRBP domain 2 (EcR2) as the uveitogenic domain. Histopathological examination indicated that EcR2-induced disease was of a chronic, non-destructive nature in the Biozzi ABH. Using synthetic overlapping peptides corresponding to EcR2, a uveitogenic and immunogenic epitope was identified corresponding to human IRBP511-530. Non-obese diabetic (NOD) mice share the same MHC class II (H-2A(g7)) molecule as the Biozzi ABH, and were also found to be susceptible to EAU induction with EcR2. This study has identified a novel mouse model of EAU, whereby disease is of a chronic, non-destructive nature, which has potential to be used in immune manipulation and neuroprotection studies. PMID- 11498257 TI - Rhesus macaque brain microvessel endothelial cells behave in a manner phenotypically distinct from umbilical vein endothelial cells. AB - Activation of endothelium is a critical step in leukocyte recruitment to the CNS and in development of neurological diseases, such as HIV-associated dementia. Due to limited availability of early disease course data, it is important to develop in vitro models of the blood-brain barrier (BBB) that can be used to address these early events. No such model of the BBB has been established for the macaque. Here, we characterize rhesus microvascular brain endothelial cells (MBEC), comparing them with rhesus umbilical vein endothelial cells (RUVEC), and discuss their suitability for future use in developing in vitro models of simian immunodeficiency virus (SIV) neuropathogenesis. We conclude that MBEC are distinct from RUVEC with respect to growth characteristics, culture requirements, morphology and expression of surface molecules important for leukocyte adhesion and immune activation. PMID- 11498258 TI - Protection against experimental autoimmune encephalomyelitis by a proteasome modulator. AB - The capacity of interferon beta to alter the course of multiple sclerosis has promoted a new therapeutic concept, based upon the modulation of the immune response rather than its suppression. As the proteasome plays a crucial role in the control of the inflammatory process and immune cell survival, targeting the proteasome appears as a novel approach for the prevention and treatment of inflammatory autoimmune diseases. We have previously shown that ritonavir, an HIV 1 protease inhibitor used in AIDS therapy, can modulate the proteasome function by inhibiting the chymotrypsin-like activity and enhancing the trypsin-like activity. We have, therefore, explored its therapeutic potential on experimental autoimmune encephalomyelitis (EAE), an experimental model of multiple sclerosis, in Lewis rats and SJL mice. Daily administration of ritonavir during autoimmune antigen stimulation prevented clinical symptoms of EAE in a dose- and time dependent manner. This protection was accompanied by an inhibition of the mononuclear cell infiltration into the central nervous system usually observed in EAE. Despite a complete absence of clinical symptoms during first EAE induction, ritonavir-treated animals became resistant to further induction of EAE, suggesting an immune mechanism of protection. These results suggest that proteasome modulation using ritonavir or analogues may be of interest for patients with multiple sclerosis. PMID- 11498259 TI - The involvement of glutamate in recall of the conditioned NK cell response. AB - The molecular mechanisms responsible for the conditioned enhancement of natural killer (NK) cell activity were investigated. The primary goal of the study was to examine the roles of glutamate and gamma-aminobutyric acid (GABA) in recall of the conditioned NK cell response. Both neurochemical blocking assay and high performance liquid chromatography (HPLC) technique were used in the study. Results from the neurochemical blocking assay demonstrated that glutamate but not GABA was required in recall of the conditioned NK cell response. NMDA but not the kainate/AMPA receptors, are believed to be involved. The levels of glutamate that were released and/or taken up also appeared to be critical in that interruption of glutamate release and/or uptake blocked the conditioned NK cell response. Results from the HPLC analysis, however, did not show any significant difference in the glutamate content between the conditioned and control brains. PMID- 11498260 TI - Astrocytes, not microglia, are the main cells responsible for viral persistence in Theiler's murine encephalomyelitis virus infection leading to demyelination. AB - The BeAn strain of Theiler's murine encephalomyelitis virus (TMEV) persists in the CNS and produces a chronic inflammatory demyelinating disease that is an animal model for human multiple sclerosis (MS). The mechanisms leading to TMEV induced demyelination are still under study but most likely involve both immune mediated and virus induced damage to cells in the CNS, both depending on viral persistence. It is therefore important to identify the cells in which continued virus production is permitted. In this study, we looked at virus infection in primary astrocytes, microglia and oligodendrocytes, derived from brains of neonatal susceptible SJL/J mice. As evidenced by Western blots and immunocytochemistry, we were able to detect viral antigens in all these brain derived cells. In addition, we extended the study to spinal cord tissues from mice suffering TMEV-induced disease. Immunohistochemistry staining with anti-TMEV sera and antibodies to specific cell markers detected viral antigens in all these cells. We then asked the question whether viral antigen present in these cells, particularly in microglia/macrophages, represented true viral replication or not. By using different techniques, including immunoprecipitation experiments and the very sensitive method of negative RNA detection through RNase protection assay, we show that both astrocytes and oligodendroglia permit de novo viral replication and viral protein synthesis but with only minimal cytopathic effects. Of these two cell types, astrocytes carry the brunt of viral replication. In microglia, on the other hand, viral replication is restricted since only minimal amounts of negative RNA copies can be demonstrated, while there are clear signs that some of these cells undergo apoptosis. These findings show that the main cell for viral replication is the astrocyte, rather than the microglia/macrophage. Most of the viral antigen present in macrophages, therefore, is probably the result of phagocytosis, rather than actual viral replication. In view of the demonstrated presence of viral replication in astrocytes and of great amounts of viral antigens in microglia/macrophages, it is possible that both types of cells act as antigen presenting cells during this immunopathological disease. PMID- 11498261 TI - HIV-1 Gp120 protein modulates corticotropin releasing factor synthesis and release via the stimulation of its mRNA from the rat hypothalamus in vitro: involvement of inducible nitric oxide synthase. AB - In the present study, we examined whether the human immunodeficiency virus type I (HIV-I) gp120 coat protein can modulate corticotropin releasing factor (CRF) secretion by using the incubation of rat hypothalamic explants as an in vitro model. Treatment of the hypothalamic fragments with recombinant gp120 resulted in a time- and concentration-dependent increase in CRF release. The maximal dose of 10 nM gp120 increased CRF release by 56.4% after 1 h, and 78.4% after 3 h, as compared with their respective controls. The intra-hypothalamic amount of CRF was also increased by 54.7% and 77.3% vs. controls after 1 and 3 h, respectively. Moreover, the action of gp120 was blocked by pretreatment with cycloheximide, suggesting that the viral protein modulates CRF secretion via an increase in its synthesis. We also investigated the effects of gp120 on CRF gene expression. RNase protection analyses of total RNA isolated from the explants indicated that 10 nM gp120 significantly increases CRF mRNA in a time-dependent manner. Furthermore, gp120 did not modify CRF mRNA stability, suggesting that the viral protein modulates CRF gene expression at the transcriptional level. Analysis of the mechanisms that mediate gp120-induced CRF synthesis was conducted. The incubation of the explants with recombinant interleukin-1 (IL-1) type I receptor antagonist (hrIL-1 ra) did not antagonize the actions of gp120 at 1 and 3 h, indicating that the effect of the latter is independent of IL-1 mediated mechanisms. The involvement of some second messenger pathways was also investigated. Specific inhibitors of cAMP-PKA, cyclo-oxygenase or heme oxygenase pathways failed to antagonize the gp120-induced increase in CRF production. By contrast, incubation with nonselective inhibitors of nitric oxide synthase (NOS), L-NAME and L-NNA, or aminoguanidine (AG), a selective inhibitor of inducible NOS (iNOS), blocked CRF release and, AG, its mRNA accumulation, stimulated by gp120, whereas selective inhibitors of endothelial and neuronal NOS had no effect. In addition, only L-NAME, L-NNA and AG were able to inhibit the gp120-stimulated production of nitrites. These results indicate that gp120 directly stimulates CRF gene expression and peptide synthesis from the rat hypothalamus in vitro via the activation of iNOS. Therefore, the actions of this viral protein on the HPA axis may, in part, reflect its ability to modulate CRF synthesis. PMID- 11498262 TI - Chronic morphine potentiates the inflammatory response by disrupting interleukin 1beta modulation of the hypothalamic-pituitary-adrenal axis. AB - Interleukin-1-beta (IL-1beta) can promote inflammation by up-regulating vascular adhesion molecules and inhibit inflammation by activating the hypothalamic pituitary-adrenal (HPA) axis to produce anti-inflammatory glucocorticoids. In this study, chronic morphine was shown to suppress IL-1beta-induction of corticotropin releasing factor (CRF) mRNA and plasma corticosterone levels. Leukocyte-endothelial adhesion (LEA) in rat mesenteric venules increased during IL-1beta- and FMLP-induced inflammation. Chronic morphine potentiated the LEA response to either IL-1beta or FMLP alone, and greatly enhanced LEA in response to combined IL-1beta and FMLP. Thus, it appears that chronic morphine exposure may promote a potentially damaging inflammatory reaction by disrupting the balance between IL-1beta-mediated local inflammation and the anti-inflammatory effects of the HPA axis. PMID- 11498263 TI - Active MRI lesion appearance in MS patients is preceded by fluctuations in circulating T-helper 1 and 2 cells. AB - BACKGROUND: The role of T cell subpopulations and their ability to produce immunoregulatory cytokines has been extensively studied in multiple sclerosis (MS). However, the exact mechanisms by which T cells and cytokines contribute to disease activity remain to be clarified. OBJECTIVES: To analyze the longitudinal relation between markers of T cell activation and differentiation and disease activity in MS patients. METHODS: During a period of 9 months, clinical disease activity was scored, monthly MRI scans were performed, and blood was taken for immune measurements in a group of 13 untreated clinically definite MS patients. RESULTS: Disease activity, as measured by the occurrence of active MRI lesions, is associated with a significant transient decrease in both T cells producing interferon-gamma (IFN-gamma) and T cells producing interleukin (IL)-4. CONCLUSIONS: Our results suggest that MRI-documented disease activity is associated with a transient decrease in circulating cytokine producing T cells, possibly due to the migration of activated T cells into the CNS. PMID- 11498264 TI - Genetic polymorphisms of IL-1beta and IL-1 receptor antagonist in association with multiple sclerosis in Japanese patients. AB - In the present study, we have investigated the association of specific polymorphisms of the interleukin (IL)-1beta and IL-1 receptor antagonist (ra) gene with both the susceptibility to and the clinical characteristics of multiple sclerosis (MS) in Japanese patients. We collected and analyzed DNA from 98 MS patients and 104 healthy controls for distribution of IL-1beta or IL-1ra polymorphisms. Our results show no significant differences in the distribution of the polymorphisms between MS patients and controls. Furthermore, no association was observed between IL-1beta or IL-1ra polymorphisms and clinical characteristics, such as age at disease onset, clinical course and severity. Together, our findings suggest that IL-1beta or IL-1ra gene polymorphisms may not be relevant in the susceptibility to MS or the clinical characteristics of Japanese patients with MS. PMID- 11498265 TI - No association of three polymorphisms in the alpha-2-macroglobulin and lipoprotein related receptor genes with multiple sclerosis. AB - Alpha-2-macroglobulin (A2M) is a proteinase inhibitor involved in deactivation of cytokines and modulation of antigen-mediated immune responses. Based on its role in inflammatory and neurodegenerative disorders, we investigated the role of A2M and its receptor low-density lipoprotein receptor-related protein (LRP) for the development of multiple sclerosis (MS). We analyzed the frequency of two polymorphisms in the A2M (Val 1000 Ile, Exon 18 del), and one polymorphism in the LRP (A216V) gene in a case control study involving 326 MS patients, and 290 controls, all defined for the expression of HLA-DR15. No association was found for any of the three polymorphism with MS. Furthermore, no differences in serum A2M levels were detected between MS patients and controls. The results do not suggest a contribution of A2M and LRP to the development of MS. PMID- 11498266 TI - Threshold of toxicological concern for chemical substances present in the diet. Report of a workshop, 5-6 October 1999, Paris, France. PMID- 11498268 TI - Assessment of the no-observed-adverse-effect level (NOAEL) of gallic acid in mice. AB - Gallic acid is a naturally occurring plant phenol obtained by the hydrolysis of tannins and is known to show some pharmacological activities. The purpose of this paper is to establish the safety of gallic acid in mice. In this study, acute administration of gallic acid even at a dose as high as 5 g/kg body weight did not produce any signs of toxicity or mortality. In the subacute study, gallic acid at a dose of 1000 mg/kg body weight did not significantly alter the hematological parameters. Further, no appreciable change was noted in the various biochemical parameters such as SGOT and SGPT, as well as many serum constituents such as protein, cholesterol, urea and bilirubin. Therefore, from this study, it may be concluded that gallic acid is non-toxic up to a level of 5000 mg/kg body weight, when given orally. In addition, the subacute study indicated the absence of cumulative toxicity, as reflected by the non-significant alterations in the parameters investigated. The NOAEL was 5000 mg/kg body weight, the highest dose tested. PMID- 11498267 TI - Induction of cytochrome P450 and/or detoxication enzymes by various extracts of rosemary: description of specific patterns. AB - The ability of rosemary to modulate cytochrome P450 (CYP) and detoxication enzymes in rat liver was evaluated by comparing the effects of dried leaves and leaf extracts with different chemical compositions: essential oil (EO) containing monoterpenes, a dichloromethane extract (DCME) containing phenolic diterpenes and a water-soluble extract (WSE) containing phenolic compounds such as rosmarinic acid and flavonoids. Chemical analyses were done in order to characterize the composition of extracts. Male Wistar rats received the leaves or extracts of rosemary in their diet at 0.5% (w/w) for 2 weeks. The effects of such treatments were evaluated for CYP (1A, 2B, 2E1), glutathione S-transferase (GST), NAD(P)H: quinone reductase (QR) and UDP-glucuronosyltransferase (UGT) activities and on protein levels (immunoblot analyses). Expression of specific UGT isoforms (mRNA semi-quantification by RT-PCR) was measured. Our study reports that EO selectively induced CYP, particularly CYP2B. WSE enhanced both CYP and detoxication enzymes. DCME acted as a monofunctional inducer, inducing GST, QR and UGT, in particular UGT1A6. Considering the specific pattern of induction obtained with DCME and WSE treatment, it should be relevant to evaluate the chemopreventive potency of these extracts on carcinogenesis in animal models. PMID- 11498269 TI - Reproduction studies in the rat with shea oleine and hardened shea oleine. AB - Shea oleine is an oil fraction derived from the nut of the tree Butyrospermum parkii, which grows in central and western Africa. There are several uses of shea oleine including its use as a frying oil and, after hardening, in margarine and toffee fat. This investigation was performed to examine the toxicity of 7 or 15% hardened shea oleine in comparison with 7 or 15% unhardened shea oleine and various commercially available materials, sheanut and palm oils, cocoa butter and toffee powder following dietary administration to rats during pre-mating, mating, pregnancy and offspring weaning in two separate investigations. Reproduction was assessed using number of litters and pups born plus survival and body weights at birth and at weaning on day 21. Skeletal evaluation using X-ray, clinical pathology and a macroscopic examination were also performed for F1 rats. Study measures for parent animals comprised evaluation of body weight, food consumption, clinical pathology, organ weights and macroscopic examination. Fatty acids and hydrocarbon levels were measured and an evaluation for lipogranulomata was made for various tissues. Results showed that shea oleine, whether unhardened or hardened, produced no evidence of reproduction toxicity and gave a similar profile to the other commercially available materials used in this study in the rat. Minor findings with shea oleine were not related to reproduction performance but comprised slightly reduced body weight gain and reduced cholesterol and raised alkaline phosphatase levels. None of the findings in this study were considered to be of toxicological significance. Thus, no evidence of reproduction toxicity was seen for both unhardened and hardened shea oleine in this investigation in the rat at levels equating to greater than 7.5 g/kg/day. PMID- 11498270 TI - The pharmacokinetics of diethanolamine in Sprague-Dawley rats following intravenous administration. AB - In order to better understand the potential toxicity of diethanolamine (DEA) and preparatory to physiologically-based pharmacokinetic model development, the pharmacokinetics of DEA at high and low internal dose through 96-h post-dosing were determined in female Sprague-Dawley rats administered 10 or 100 mg/kg uniformly labeled 14C-DEA via intravenous injection. Clearance of DEA from blood was calculated to be approximately 84 ml/h/kg at the low dose, increasing to approximately 242 ml/h/kg at the high dose. The primary route of excretion of administered radioactivity, approximately 25-36%, was via the urine as parent compound. A majority of the administered radioactivity was recovered in the tissues of treated rats, especially in the liver and kidneys, suggesting a propensity of DEA or its metabolites for bioaccumulation. An accumulation of radioactivity also occurred gradually in the red blood cells from about 6-96 h post-dosing. Some evidence of dose dependency in the fate of iv-administered DEA was observed, suggesting that saturation of the bioaccumulation process(es) occurred at a dose level of 100 mg/kg. PMID- 11498271 TI - Induction of sister chromatid exchanges by 2,4-dichlorophenoxyacetic acid in somatic and germ cells of mice exposed in vivo. AB - 2,4-dichlorophenoxyacetic acid (2,4-D) is one of the most widely used selective herbicides throughout the world; however, the studies that have been conducted to establish its genotoxic potential have given conflicting results. The aim of this investigation was to determine whether the herbicide increases the frequency of sister chromatid exchanges (SCEs) in bone marrow and spermatogonial cells of mice exposed in vivo. The experiment included an oral administration of 2,4-D to three groups of mice (50,100 and 200 mg/kg), as well as to a control group of animals administered with distilled water, pH 10.5 and another group injected with cyclophosphamide (50 mg/kg). In somatic cells, the results showed a significant SCE increase with the two high doses tested, a response that was manifested in a dose-dependent manner. With regard to the mitotic index and the cell proliferation kinetics, there were no modifications exerted by 2,4-D; however, cyclophosphamide induced cytotoxic damage and a cell-cycle delay. With respect to the germ cells, the genotoxic results were similar to those described earlier; that is, there was a significant SCE increase induced by the two high 2,4-D doses tested and a higher genotoxic damage was observed in the animals treated with cyclophosphamide. Our investigation established that 2,4-D is a moderate genotoxicant in mice treated in vivo with high doses, and suggests a minor hazard for humans in the present conditions of its use. PMID- 11498272 TI - Effects of toxaphene on the immune system of cynomolgus (Macaca fascicularis) monkeys. AB - Toxaphene, dissolved in glycerol/corn oil, was administered at 0.1, 0.4 or 0.8 mg/kg body weight/day in gelatin capsules to groups of 10 young adult female cynomolgus monkeys (Macaca fascicularis), while a group of five male monkeys (Macaca fascicularis) received 0.8 mg/kg body weight/day. Control male (a group of five) and female (a group of 10) monkeys ingested the glycerol/corn oil vehicle only. Treatment continued for 75 weeks. Testing for immune effects was initiated at 33 weeks of treatment. Immunization was initiated at 44 weeks of treatment. Pairwise comparisons between each of the treated female groups to the control indicated that the mean primary (post-immunization weeks 1-4) and secondary (post-immunization weeks 5-8) anti-SRBC IgM responses were significantly reduced at the 0.4 and 0.8 mg/kg body weight/day doses compared to the control (P< or =0.05). The mean primary (post-immunization weeks 1-4) anti SRBC IgG response was significantly reduced compared to the control (P< or =0.05), while the secondary (post-immunization weeks 5-8) anti-SRBC IgG was not significantly affected by treatment (P>0.05). The mean anti-tetanus toxoid IgG response in the 0.8 mg/kg body weight/day dose group The mean primary anti-SRBC (IgM) response in the treated males was significantly different from the control (P<0.05), while the primary anti-SRBC IgG response was not affected by treatment. The mean absolute B-lymphocyte numbers in the female group administered 0.8 mg/kg of toxaphene was significantly reduced compared to the control (P< or =0.05). All other parameters including the natural killer cell activity, the delayed-type hypersensitivity response, the lymphoproliferative response of peripheral blood leukocytes to the mitogens Con A and PWM and the serum cortisol levels were not affected significantly by treatment (P>0.05). The no-observed-adverse-effect level (NOAEL) for the female monkeys based on the toxaphene effects on humoral immunity was 0.1 mg/kg body weight/day. PMID- 11498273 TI - Processing procedures of brick tea and their influence on fluorine content. AB - China is the only country that produces brick tea, and more than 90% of the brick tea is consumed in the western minority nationality regions of China. The high fluorine content of brick tea is possibly associated with the special processing procedures, but no investigation has been conducted in this field. To explore the characteristic features of brick tea manufacturing and the alterations in fluorine content during the processing procedures, we performed a field survey involving two brick tea factories and the nearby tea plantations. For the fluorine contents of the initial, intermediate and final processing products, altogether eight types of specimens were collected and determined by using the ion-selective electrode standard curve technique. It was found that the raw material tea leaf for brick tea processing was old, coarse and not the tender delicate tea leaf used for ordinary green or black tea processing. For the fluorine content of the raw material tea leaf, the intermediate and the final products showed that the fresh raw leaf contained a fluorine content as high as 489.31-512.68 mg/kg. During one fermentation-like processing procedure, the fluorine content rose by 4.67% and 1.88% in the specimens from the two factories, respectively, which revealed no statistical significance (P>0.5). These results suggest that the high fluorine content in brick tea might be due to the high content in the raw material and not related with the processing procedures. PMID- 11498275 TI - Diversity of endotoxin-induced nitrotyrosine formation in macrophage-endothelium rich organs. AB - The administration of bacterial lipopolysaccharide (LPS; endotoxin) can stimulate the development of the systemic inflammatory response syndrome, which can compromise the function of many organ systems, resulting in multiple organ failure. Activation of macrophages and cytokines by endotoxin and the subsequent formation of reactive oxygen and nitrogen species are of central pathogenic importance in various inflammatory diseases including sepsis. However, whether different tissues behave the same in pathological changes produced by LPS and what factors may affect pathological processes and protein tyrosine nitration in different organs, still remain to be evaluated. In the present study, we investigated the distribution of nitrotyrosine and other pathological changes induced by LPS in rat liver, spleen, and lung, all of which are rich in macrophages and endothelial cells. Our study revealed two important findings: first, a denitration activity in spleen white pulp might play a key role to protect the areas from nitration. Similar activity might also exist in endothelial cells of sinusoids and capillaries. Second, protein nitration might not induce significant tissue damage as shown in liver and spleen. However, inflammatory infiltration with increased formation NO* and other reactive species may result in severe tissue injury, as demonstrated in lung after LPS administration. PMID- 11498276 TI - A role for free radicals and nitric oxide in delayed recovery in aged rats with chronic constriction nerve injury. AB - Using a reversible chronic constriction injury (CCI) model of neuropathic pain, we previously demonstrated that changes in thermal hyperalgesia correlate with the changes in peripheral microvascular blood flow in the affected paw, and that recovery can be assessed by normalization of both behavioral and vascular responses. Using the same model, this study examined age-related changes in recovery after nerve injury and the involvement of free radicals and nitric oxide (NO) in these changes. Four loose, nonconstrictive ligatures were applied to the sciatic nerve in the right, mid-thigh region of young and old (3 and 24 months) Sprague Dawley rats. All rats were monitored weekly (for 8-10 weeks) for their thermal threshold using a 46 degrees C water bath and some groups were used to examine endothelial and smooth muscle-dependent microvascular responses to substance P (SP) and sodium nitroprusside (SNP), respectively. These substances were perfused over the base of blisters raised on the footpad innervated by the injured nerve. Free radical activity in the sciatic nerve was assessed by measuring the activity of xanthine oxidase (XO) and lipid hydroperoxides (LPO). Young rats showed signs of recovery (reduction in thermal hyperalgesia and improvement of peripheral microvascular blood flow) from the fifth week. No signs of recovery were observed in old rats for 8 weeks, with some reduction in thermal hyperalgesia observed by weeks 9 and 10. XO activity was significantly higher in young injured nerves compared to sham (400%) and was even significantly greater in old injured nerves (680%). Similarly, old injured nerves showed 300% increase in LPO levels compared to sham. The role of reactive oxygen species (ROS) in delayed recovery in old rats was examined using the antioxidant tirilazad mesylate. Tirilazad (20 mg/kg) was injected intramuscularly (im) in the mid-thigh region starting on day 1 post CCI, (early treatment) or day 7 (late treatment). Levels of LPO in the injured sciatic nerves were significantly reduced using either early or late treatment, however tirilazad had opposing effects on recovery, prolonging or alleviating thermal hyperalgesia, respectively. The role of neuronal nitric oxide (nNO) was then examined using the specific neuronal nitric oxide synthase (nNOS) inhibitor, 3-bromo-7-nitroindazole (3Br-7NI) (10 mg/kg). 3Br-7NI resulted in a significant alleviation of thermal hyperalgesia with improvement in the vascular responses from weeks 5 and 6 onwards. A combination of 3Br-7NI and tirilazad treatment was also used but did not show an additive effect. The results suggest that ROS and nNO contribute to delayed recovery of injured nerves in old rats and to the maintenance of thermal hyperalgesia and the reduction in microvascular blood flow in the area innervated by the injured nerve. The results also raise the notion that possible interaction of free radicals with NO to form peroxynitrite might be responsible for such delayed recovery. Ironically, this study also reveals a positive role for free radicals in tissue repair and raises the notion that early intervention with antioxidants could exert a negative effect on repair of injured nerves. PMID- 11498277 TI - Peroxynitrite-modified 99mTc-beta-VLDL: tissue distribution and plasma clearance rate. AB - Free radicals superoxide (O(2)(-)) and nitric oxide (*NO) are generated by blood vessels and can rapidly react to produce a peroxynitrite anion (ONOO(-)), a powerful oxidant that modifies lipoproteins making them more atherogenic. The aim of this study was to investigate the effect of peroxynitrite-induced modifications on beta-very-low-density lipoprotein (beta-VLDL) as to its biodistribution and plasma clearance rate, as well as the uptake of these particles by THP-1 cells. After being injected into New Zealand White rabbits, the peroxynitrite-modified beta-VLDL (99mTc-per-beta-VLDL) was cleared from circulation faster than the native beta-VLDL (99mTc-nat-beta-VLDL) in both normocholesterolemic rabbits (NC) and in hypercholesterolemic rabbits (HC). In HC rabbits, the fractional clearance of 99mTc-labeled beta-VLDL was significantly lower than in NC rabbits. The in vivo studies showed that accumulation of 99mTc labeled beta-VLDL, expressed per gram of tissue, followed the decreasing order: kidney > liver > spleen > adrenal gland >or= lung > aortic arch > heart >or= abdominal aorta > thoracic aorta > psoas muscle. The high accumulation in the kidneys suggests the processing of 99mTc-labeled apolipoproteins by receptors present in kidney cells. The accumulation of 99mTc-nat-beta-VLDL in the whole organ was the following: liver > kidney > heart > spleen > adrenal gland > aorta in HC and NC rabbits. The uptake of 99mTc-per-beta-VLDL by the spleen was greater than the uptake by the heart in both groups. The in vitro studies showed that the uptake of 99mTc-per-beta-VLDL by THP-1 cells was higher than that of 99mTc-nat beta-VLDL. These results show that peroxynitrite-modified beta-VLDL is rapidly removed from plasma and accumulates in several tissues, mainly in the liver and kidney. This may be particularly important in hypercholesterolemic situations that could favor the accumulation of native and peroxynitrite-modified beta-VLDL in several tissues. PMID- 11498278 TI - Lipopolysaccharide and interferon-gamma-induced nitric oxide production and protein oxidation in mouse peritoneal macrophages are affected by glutathione peroxidase-1 gene knockout. AB - This study investigated the role of glutathione peroxidase-1 (GPX1) in protein oxidation in peritoneal macrophages. Macrophages isolated from both wild-type (WT) and GPX1 knockout (KO) mice were activated by lipopolysaccharide (LPS, 1 microg/ml) and interferon-gamma (IFN, 10 U/ml for 24 or 48 h in the presence or absence of 1 microM diquat (DQ), 250 microM aminoguanidine (AG, an inhibitor of inducible nitric oxide synthase), and (or) 100 microM diethyldithiocarbamate (DETC, an inhibitor of Cu,Zn-SOD). In the KO macrophages, there was no protein band detected by Western blot with anti-GPX1 antibody and 98% reduction in total GPX activity compared with WT cells. Nitric oxide (NO) synthesis was greatly enhanced after 24 h by GPX1 knockout and DQ, but inhibited by AG or DETC. Protein carbonyl formation in total cell extract was clearly associated with NO synthesis as higher levels of protein carbonyl were detected in activated KO than WT macrophages, and DQ enhanced slightly while AG or DETC virtually blocked its formation. A similarly marginal effect of GPX1 KO was observed on protein nitration. The LPS/IFN/DQ-induced DNA fragmentation was blocked by AG, but not by DETC. Cell viability at 48 h was decreased by the LPS/IFN activation and further reduced by the addition of DQ, but restored by AG. In conclusion, GPX1 affects the NO production in activated peritoneal macrophages and protects these cells against NO-associated protein oxidation. PMID- 11498279 TI - Association of tocopherols with circulating autoantibody levels against an oxidized DNA nucleoside in humans. AB - Autoantibodies against oxidized DNA bases are found in vivo and have been used as an indicator of oxidative damage, yet little is known concerning their individual variation and relation to serum micronutrients. Human plasma anti-5-hydroxymethyl 2'-deoxyuridine (HMdU) autoantibody (aAb) levels were repeatedly determined in 41 women and 11 men, and found to have small within-individual variation over time, but large between-individual differences. A positive association in both women (r = .5762, p = .0001) and men (r = .415, p = .2) between plasma total tocopherols and antibody levels was observed. Autoantibody levels were lower in postmenopausal women (8.37 +/- 1.61 vs. 17.18 +/- 2.85 in premenopausal women, p < .01), independently of plasma tocopherol. However, aAb titers in postmenopausal women were still significantly associated with plasma tocopherol levels and adjustment for menopausal status in women yielded a highly significant correlation between HMdU aAb levels and total tocopherol (r = .7342, p = .0001). Plasma malondialdehyde equivalents (MDA), a measure of lipid peroxidation, were also higher in individuals with either high plasma alpha-tocopherol or high beta+gamma-tocopherol levels. The positive association of tocopherols with markers of oxidative damage may reflect a response to the generation of endogenous oxidants associated with enhanced immune function. The decrease in aAb level in postmenopausal women may similarly reflect decreased immune function associated with decreased estrogen levels. PMID- 11498280 TI - Superoxide-mediated early oxidation and activation of ASK1 are important for initiating methylglyoxal-induced apoptosis process. AB - Methylglyoxal (MG) is a physiological metabolite, but it is known to be toxic, inducing stress and causing apoptosis. Our previous studies demonstrated that MG induced apoptosis in Jurkat cells by activating the c-Jun N-terminal kinase (JNK) signal transduction pathway, which induced an obvious decrease in mitochondrial membrane potential, followed by caspase-3 activation. Here, we observed that MG induced apoptosis was associated with both rapid production of superoxide anion (O(2)(-)) followed by a marked increase in ROS and striking and temporal activation of ASK1. Overexpression of wild-type ASK1 could enhance the rate of apoptosis induced by MG, whereas the expression of the kinase-inactive form of ASK1 notably prevented cells from MG-induced death. NAC and PDTC blocked the activation of ASK1 and MG-induced apoptosis completely. Moreover, nonthiol antioxidants SOD-mimic MnTBAP and catalase together obviously inhibited MG induced ASK1 activation and apoptosis induction. Correspondingly, MG-mediated ASK1 activation was enhanced by diethyldithiocarbamate (DDC). Addition of antioxidant into the culture of cells at a later stage (4-8 h after the initial MG treatment) failed to prevent their death. These results suggest that activating ASK1 at the early stage linking to production of O(2)(-) is crucial for subsequent progression of apoptosis in MG-treated Jurkat cells. PMID- 11498281 TI - Se-methylselenocysteine induces apoptosis mediated by reactive oxygen species in HL-60 cells. AB - Recent studies have implicated apoptosis as one of the most plausible mechanisms of the chemopreventive effects of selenium compounds, and reactive oxygen species (ROS) as important mediators in apoptosis induced by various stimuli. In the present study, we demonstrate that Se-methylselenocysteine (MSC), one of the most effective selenium compounds at chemoprevention, induced apoptosis in HL-60 cells and that ROS plays a crucial role in MSC-induced apoptosis. The uptake of MSC by HL-60 cells occurred quite early, reaching the maximum within 1 h. The dose dependent decrease in cell viability was observed by MSC treatment and was coincident with increased DNA fragmentation and sub-G(1) population. 50 microM of MSC was able to induce apoptosis in 48% of cell population at a 24 h time point. Moreover, the release of cytochrome c from mitochondria and the activation of caspase-3 and caspase-9 were also observed. The measurement of ROS by dichlorofluorescein fluorescence revealed that dose- and time-dependent increase in ROS was induced by MSC. N-acetylcysteine, glutathione, and deferoxamine blocked cell death, DNA fragmentation, and ROS generation induced by MSC. Moreover, N-acetylcysteine effectively blocked caspase-3 activation and the increase of the sub-G(1) population induced by MSC. These results imply that ROS is a critical mediator of the MSC-induced apoptosis in HL-60 cells. PMID- 11498282 TI - Intracellular iron, but not copper, plays a critical role in hydrogen peroxide induced DNA damage. AB - The role of intracellular iron, copper, and calcium in hydrogen peroxide-induced DNA damage was investigated using cultured Jurkat cells. The cells were exposed to low rates of continuously generated hydrogen peroxide by the glucose/glucose oxidase system, and the formation of single strand breaks in cellular DNA was evaluated by the sensitive method, single cell gel electrophoresis or "comet" assay. Pre-incubation with the specific ferric ion chelator desferrioxamine (0.1 5.0 mM) inhibited DNA damage in a time- and dose-dependent manner. On the other hand, diethylenetriaminepentaacetic acid (DTPA), a membrane impermeable iron chelator, was ineffective. The lipophilic ferrous ion chelator 1,10 phenanthroline also protected against DNA damage, while its nonchelating isomer 1,7-phenanthroline provided no protection. None of the above iron chelators produced DNA damage by themselves. In contrast, the specific cuprous ion chelator neocuproine (2,9-dimethyl-1,10-phenanthroline), as well as other copper-chelating agents, did not protect against H(2)O(2)-induced cellular DNA damage. In fact, membrane permeable copper-chelating agents induced DNA damage in the absence of H(2)O(2). These results indicate that, under normal conditions, intracellular redox-active iron, but not copper, participates in H(2)O(2)-induced single strand break formation in cellular DNA. Since BAPTA/AM (1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester), an intracellular Ca(2+) chelator, also protected against H(2)O(2)-induced DNA damage, it is likely that intracellular Ca(2+) changes are involved in this process as well. The exact role of Ca(2+) and its relation to intracellular transition metal ions, in particular iron, needs to be further investigated. PMID- 11498283 TI - Influence of age on activities of antioxidant enzymes and lipid peroxidation products in erythrocytes and neutrophils of Down syndrome patients. AB - Thirty-seven individuals with Down syndrome (DS) were divided into four age categories: (i) 1 to < 6 years, (ii) 6 to < 13 years, (iii) 13 to < 20 years, and (iv) over 20 years. Activities of antioxidant enzymes found in individual age categories were different, but the differences between age groups were not statistically significant. We confirmed significantly higher activities of Cu/Zn superoxide dismutase (SOD) and glutathione peroxidase (GPx) in blood cells of people with DS as compared to 35 controls, which consisted, for the first time, of siblings of children with DS. No significant differences were found in activities of catalase and glutathione reductase in DS vs. controls. A significant difference was observed in serum concentration of malondialdehyde (MDA) in DS vs. controls (8.39 +/- 0.34 micromol/l vs. 7.34 +/- 0.27 micromol/l; p = .021) and concentration of MDA in erythrocytes of individuals with DS between the third and fourth age group (p = .05). In DS persons, an elevated ratio of SOD to catalase plus GPx with respect to the controls in all age categories was found, suggesting oxidative imbalance, potentially contributing to accelerated aging observed in these persons. PMID- 11498284 TI - Oxidative stress modulates osteoblastic differentiation of vascular and bone cells. AB - Oxidative stress may regulate cellular function in multiple pathological conditions, including atherosclerosis. One feature of the atherosclerotic plaque is calcium mineral deposition, which appears to result from the differentiation of vascular osteoblastic cells, calcifying vascular cells (CVC). To determine the role of oxidative stress in regulating the activity of CVC, we treated these cells with hydrogen peroxide (H(2)O(2)) or xanthine/xanthine oxidase (XXO) and assessed their effects on intracellular oxidative stress, differentiation, and mineralization. These agents increased intracellular oxidative stress as determined by 2,7 dichlorofluorescein fluorescence, and enhanced osteoblastic differentiation of vascular cells, based on alkaline phosphatase activity and mineralization. In contrast, H(2)O(2) and XXO resulted in inhibition of differentiation markers in bone osteoblastic cells, MC3T3-E1, and marrow stromal cells, M2-10B4, while increasing oxidative stress. In addition, minimally oxidized low-density lipoprotein (MM-LDL), previously shown to enhance vascular cell and inhibit bone cell differentiation, also increased intracellular oxidative stress in the three cell types. These effects of XXO and MM-LDL were counteracted by the antioxidants Trolox and pyrrolidinedithiocarbamate. These results suggest that oxidative stress modulates differentiation of vascular and bone cells oppositely, which may explain the parallel buildup and loss of calcification, seen in vascular calcification and osteoporosis, respectively. PMID- 11498285 TI - v-Ha-RaS oncogene upregulates the 92-kDa type IV collagenase (MMP-9) gene by increasing cellular superoxide production and activating NF-kappaB. AB - Matrix metalloproteinase 9 (MMP-9) degrades basement membrane type IV collagen and is expressed during cellular migration and invasion. Here we show that v-Ha Ras overexpression in rat kidney epithelial cells (REC) caused upregulation of MMP-9 gene expression in part by increasing cellular oxidant levels. v-Ha-Ras mediated the production of superoxide in Ras-transfected cells, which was associated with upregulated MMP-9 gene expression. Conversely, v-Ha-Ras expression decreased steady-state levels of mRNAs from tissue inhibitor of metalloproteinase 1 (TIMP-1), an inhibitor of MMP-9; plasminogen activator inhibitor 1 (PAI-1), which indirectly activates MMP-9 by increasing plasmin levels; and collagen IV, a substrate of MMP-9 and a major component of basement membrane. Gel mobility shift assays demonstrated that Ras overexpression enhanced NF-kappaB, but not AP-1 DNA binding to motifs in the MMP-9 gene promoter. The Ras induced increase in NF-kappaB DNA binding could be inhibited by treatment with the antioxidants N-acetyl-L-cysteine and glutathione monoester, suggesting that intracellular oxidant levels can mediate MMP-9 transcription. Our findings identify an important role for Ras in the regulation of MMP-9 expression, and suggest that increased superoxide production can upregulate MMP-9 expression and thus contribute to malignant conversion. PMID- 11498286 TI - Enhanced antioxidant and cytoprotective abilities of vitamin E succinate is associated with a rapid uptake advantage in rat hepatocytes and mitochondria. AB - Numerous in vitro studies attest to the enhanced ability of vitamin E succinate (TS), as compared with conventional vitamin E compounds such as unesterified d alpha-tocopherol (T) and d-alpha-tocopheryl acetate (TA), to protect hepatocytes from toxic oxidative stress. In the present study we tested the hypothesis that this unique protective ability is related to an enhanced cellular accumulation of TS. The results of this study indicate, using both in vitro and in vivo model systems, that acute TS administration results in a rapid increase in T and TS content and antioxidant protection of hepatocytes and mitochondria. In contrast, conventional vitamin E compounds such as T and TA lack these same protective properties. We suggest that TS acts as a unique delivery system for T, rapidly accumulating in cellular and mitochondrial membranes and gradually releasing active T to prevent membrane oxidative damage. We propose that TS administration may prove useful for the prevention and treatment of oxidative stress-mediated diseases, especially those of mitochondrial origin. PMID- 11498287 TI - Further evidence that oxidative stress may be a risk factor responsible for the development of atherosclerosis. AB - There are numerous data suggesting that oxidative stress may be involved in the development of atherosclerosis. Therefore, in the present study we measured the amount of 8-hydroxy-2'-deoxyguanosine (8-OH-dG), one of the typical biomarkers of oxidative stress, in DNA isolated from lymphocytes of the patients and in the control group. Levels of antioxidant vitamins (A, C, and E) and intracellular labile iron pool (LIP), which can influence oxidative stress, were also determined. Blood samples were obtained from a control group of 55 healthy persons and from 43 atherosclerotic patients. 8-OH-dG and the vitamin levels were measured by high-performance liquid chromatography. Labile iron pool in lymphocytes was analyzed by fluorescent assay. The levels of 8-OH-dG and LIP were significantly higher and vitamin C concentration was significantly lower in the patient group than in the control group. The rest of the analyzed parameters do not significantly differ between the groups. A lower concentration of vitamin C and higher levels of labile iron pool in a group of atherosclerotic patients when compared with the control group may lead to oxidative stress, which is manifested by a higher level of 8-OH-dG in blood lymphocytes. All these factors may create an environment that promotes the development of atherosclerosis. PMID- 11498288 TI - Direct effect of Taxol on free radical formation and mitochondrial permeability transition. AB - To elucidate the potential role of mitochondria in Taxol-induced cytotoxicity, we studied its direct mitochondrial effects. In Percoll-gradient purified liver mitochondria, Taxol induced large amplitude swelling in a concentration-dependent manner in the microM range. Opening of the permeability pore was also confirmed by the access of mitochondrial matrix enzymes for membrane impermeable substrates in Taxol-treated mitochondria. Taxol induced the dissipation of mitochondrial membrane potential (DeltaPsi) determined by Rhodamine123 release and induced the release of cytochrome c from the intermembrane space. All these effects were inhibited by 2.5 microM cyclosporine A. Taxol significantly increased the formation of reactive oxygen species (ROS) in both the aqueous and the lipid phase as determined by dihydrorhodamine123 and resorufin derivative. Cytochrome oxidase inhibitor CN(-), azide, and NO abrogated the Taxol-induced mitochondrial ROS formation while inhibitors of the other respiratory complexes and cyclosporine A had no effect. We confirmed that the Taxol-induced collapse of DeltaPsi and the induction of ROS production occurs in BRL-3A cells. In conclusion, Taxol-induced adenine nucleotide translocase-cyclophilin complex mediated permeability transition, and cytochrome oxidase mediated ROS production. Because both cytochrome c release and mitochondrial ROS production can induce suicide pathways, the direct mitochondrial effects of Taxol may contribute to its cytotoxicity. PMID- 11498292 TI - Immunology in Scandinavia. PMID- 11498294 TI - Parasite genomics: current status and future prospects. AB - The past year has brought great progress in the genome-sequencing efforts on a large number of protozoan and metazoan parasites. Whereas many of these projects are in their initial stages, at least one (for Plasmodium falciparum) is nearing completion. The information released to date has been most revealing with respect to immune evasion mechanisms. PMID- 11498295 TI - Th1/Th2 effector choice in parasitic infection: decision making by committee. AB - Parasitic infections frequently result in highly polarized CD4+ T cell responses characterized by dominant Th1 or Th2 cytokine production profiles. Although previously thought to be strictly dependent on signaling by the differentiative cytokines, IL-12 and IL-4, recent data indicate that this polarization may be primarily decided instead by a series of different factors intrinsic to the pathogen-antigen-presenting-cell interaction that influence T cell priming. PMID- 11498296 TI - Innate immune responses of epithelial cells following infection with bacterial pathogens. AB - The ability to discriminate between pathogenic and non-pathogenic bacteria is extremely important for epithelial cells lining mucosal surfaces and is particularly so in colonic epithelial cells. Accumulating evidence suggests that bacterial recognition systems used by epithelial cells are very different from those in cells of the myeloid lineage and are likely to have developed to maintain mucosal surfaces in a state of homeostasis with the normal microbial flora. Bacterial invasion of epithelial cells or breach of the epithelial barrier provides a signal to epithelial cells to initiate inflammatory responses, which are key events for the clearance of the infecting microbe. Therefore, elucidation of the mechanisms by which epithelial cells recognize bacteria and bacterial products, and of the nature of the innate immune responses that are triggered by these factors are important for our understanding of both the immunology of mucosal surfaces and bacterial pathogenesis. PMID- 11498297 TI - Immune responses to intracellular bacteria. AB - The multifaceted dialogue between intracellular bacteria and the mammalian host continues to be an exciting issue from both the scientific and public-health viewpoint. The recent year has witnessed some particularly impressive progress in knowledge about the two major culprits affecting the health of mankind, Mycobacterium tuberculosis and Salmonella typhi - the causative agents of tuberculosis and typhoid fever. PMID- 11498298 TI - Epitope clusters in the major outer membrane protein of Chlamydia trachomatis. AB - Immunopathology that is caused by re-infection with Chlamydia trachomatis is very common in humans despite regular responses to multiple, often conserved, antibody and T cell epitopes. Recurrent mutations that disrupt T cell epitopes in the major outer membrane protein in clinical isolates and the reduced transcription of HLA genes by infected cells may be evidence for pathogen evasion of protective immune responses. Subunit vaccines containing recently discovered clusters of T cell epitopes in the major outer membrane protein that are presented with diverse HLA allotypes may allow widespread protective immunization while avoiding the suppression of lasting immunity that occurs by unknown mechanisms associated with infection. PMID- 11498299 TI - Antigen presentation to CD8+ T cells: cross-priming in infectious diseases. AB - Recent studies indicate that, in most types of infections, antigen presentation by 'professional' bone-marrow-derived cells is essential for priming pathogen specific CD8+ T cells. This is true even in the absence of direct infection of these cells, which indicates that cross-priming is an essential component of the immune response against pathogens. PMID- 11498300 TI - Virus subversion of immunity: a structural perspective. AB - Over the past year, we have witnessed the discovery of further virus immuno evasins--proteins that alter the host immune response. Although many of these factors have been described over the past decade, the structural basis underlying their biology has lagged behind. Structural data have now been obtained for several such proteins. Major advances of the past year include the structures of a viral chemokine-binding protein, of an intact viral regulator of complement activation and of an immuno-evasin with its cellular target. PMID- 11498301 TI - Immunity to infections following hematopoietic cell transplantation. AB - Hematopoietic cell transplantation has progressed from the use of unpurified bone marrow cells or mobilized peripheral blood cells to the use of purified stem cells and progenitor cells. These kinds of transplants can be designed to provide not only hematopoietic rescue but also augmented innate and acquired immunity. PMID- 11498302 TI - NK cells and NKT cells in innate defense against viral infections. AB - NK cells contribute to innate defense during certain viral infections, but the mechanisms for their regulation and delivery of antiviral effects are incompletely understood. A second NK cell population, from within T cell populations--NKT cells--has a unique potential to initiate cellular effector mechanisms, including those delivered by NK cells, provided that the antigen for their restricted TCR is induced during infection. If elicited, particular innate cytokine responses promote activation of NK cell cytotoxicity or IFN-gamma production. These responses can contribute to defense by mediating antiviral and/or immunoregulatory effects. Roles of positive or negative receptors for target cells in protection against viruses are less clear. Exciting new data indicate that, in at least one system, NK cell receptors that positively signal for activation participate in the recruitment of these cells into antiviral defense mechanisms. Other recent evidence suggests that NKT cells may be important for protection during one viral infection and may be artificially activated by delivery of antigen to promote antiviral defense. Taken together, these recent advances in the characterization of the NK and NKT cell responses are filling in the details of the complex and critical events taking place, at the earliest times after challenge, to promote resistance to viruses. PMID- 11498303 TI - Expression and function of NK cell receptors in CD8+ T cells. AB - A wide variety of inhibitory and stimulatory NK cell receptors are expressed by some CD8+ cytotoxic T lymphocytes in mice and humans. Recent data address the induction of these receptors on activated or memory CD8+ T cells and have led to hypotheses addressing their function in the CD8+ T cell response. PMID- 11498304 TI - CD1-specific T cells in microbial immunity. AB - Recent results strengthen evidence that CD1-restricted T cells play important roles in host defense against microbial infections. Human subjects recently infected with Mycobacterium tuberculosis showed elevated responses to CD1c mediated presentation of a microbial lipid antigen, compared with control donors. Activation of CD1d-restricted NKT cells with a synthetic glycolipid antigen results in improved immune responses to several infectious pathogens. PMID- 11498305 TI - CD8+ cytotoxic T lymphocyte responses to lentiviruses and herpesviruses. AB - Immune containment of persistent viral infections has long been a focus of interest for investigators. However, the technologies needed to evaluate the role of CD8+ cytotoxic T lymphocytes (CTLs) in this process have only recently become available. Recent studies performed using tetramer, ELISPOT and cytokine production assays have evaluated the role of CD8+ CTLs in controlling lentivirus and herpesvirus infections in humans and nonhuman primates. These studies demonstrate dramatic expansions of virus-specific CTLs in primary infection and the maintenance of unexpectedly high levels of virus-specific CTLs in chronic infection. These findings underscore the importance of CD8+ CTLs in the immune control of persistent viral infections. PMID- 11498306 TI - Reconstitution of CD4+ T lymphocytes in HIV-infected individuals following antiretroviral therapy. AB - Immune reconstitution during antiretroviral therapy has recently been shown to depend upon multiple factors at work in T cell homeostasis, amongst which the reduction of thymus dysfunction and of immune hyperactivation are instrumental. The optimism that has been raised by the restoration of hosts' defenses against opportunistic pathogens is, however, balanced by the poor immunity restored against HIV; thus, innovative immune interventions are required. PMID- 11498307 TI - Vaccines for the prevention of HIV-1 disease. AB - Clinical investigation in humans and experimental lentivirus infection in nonhuman primates have advanced our understanding of immune responses that control HIV-1 disease. Recently, immunization approaches in macaques have shown that the immune response can control viremia and improve clinical outcome. When such vaccine strategies are formulated to be similarly immunogenic in humans, they could form the basis for the development of candidate AIDS vaccines that would prevent infection, suppress progression to disease or reduce HIV-1 transmission in humans. PMID- 11498308 TI - Retroviruses as tools to study the immune system. AB - Retrovirus-based vectors provide an efficient means to introduce and express genes in cells of the immune system and have become a popular tool to study immune function. They are easy to manipulate and provide stable, long-term gene expression because they integrate into the genome. Current retroviral vectors do have limitations that affect their usefulness in certain applications. However, recent advances suggest a number of ways in which these vectors might be improved to extend their utility in immunological research. PMID- 11498309 TI - The ethics of vaccine usage in society: lessons from the past. Commentary. PMID- 11498310 TI - Commercial propofol solutions: is the more expensive also the more effective? AB - STUDY OBJECTIVE: To compare the efficacy and safety of four commercial propofol solutions marketed in Israel. DESIGN: Prospective, randomized, double-blind study. SETTING: University-affiliated medical center. PATIENTS: 120 ASA physical status I and II nullipara patients undergoing dilatation and curettage for interrupted pregnancy. INTERVENTIONS: Patients were randomized into four groups of 30 patients each. Group 1 was anesthetized with Diprivan (AstraZeneca, UK), group 2 with Recofol (Leiras Oy, Finland), group 3 with Propofol (Abbott), and group 4 with Diprofol (Taro, Israel). MEASUREMENTS AND MAIN RESULTS: The four study groups were similar in mean age and weight. There were no statistically significant differences in the administered doses, quality of anesthesia, recovery time, or adverse effects among the four groups. CONCLUSIONS: Diprivan, Recofol, Diprofol, and Propofol Abbott are equally effective as anesthesia induction drugs for dilation and curettage, with a similar incidence of adverse effects. Because cost limitations have become a significant factor in medical care, the choice of drug in this group should be based solely on cost considerations. PMID- 11498311 TI - A double-blind, randomized comparison of low-dose rocuronium and atracurium in a desflurane anesthetic. AB - STUDY OBJECTIVES: To compare the neuromuscular and hemodynamic effects of rocuronium and atracurium when administered during a desflurane-based anesthetic. DESIGN: Randomized, double-blind clinical trial. PATIENTS: 51 adult ASA physical status I and II patients scheduled for general surgical operations. SETTING: University-based NCI-designated cancer center. INTERVENTIONS: Patients received either 0.45 mg/kg rocuronium (n = 28) or atracurium 0.5 mg/kg (n = 23). Induction of anesthesia was accomplished by 2 microg/kg fentanyl intravenously (IV) and 1.5 mg/kg propofol IV and maintained by a nitrous oxide/oxygen desflurane anesthetic. MEASUREMENTS AND MAIN RESULTS: A neuromuscular monitor was used at the adductor pollicis to monitor and record twitch response to train-of-four electrical stimulation. Baseline heart rate (HR) and blood pressure (BP) were measured and again at 2, 5, 10, and 15 minutes after muscle relaxant administration. Patients in the rocuronium group were found to have shorter times to 80%T(1)depression (109 +/- 53 vs. 135 +/- 47 sec), although those differences did not reach statistical significance (p = 0.07). Percent of the first twitch (T(1) ) was significantly lower in the patients receiving rocuronium at 60 seconds (53 +/- 24 vs. 73 +/- 27 sec; p = 0.006) and 90 seconds (25 +/- 22 vs. 47 +/- 29 sec; p = 0.003) than in the patients receiving atracurium. Duration was shorter in rocuronium-treated patients (25% T(1) recovery = 32 +/- 12 vs. 54 +/- 14 min; p < 0.001) than the patients receiving atracurium. Intubation scores were better at 60 seconds after muscle relaxant administration in the rocuronium group. No significant differences in HR or BP were seen between the patients in the two groups. CONCLUSIONS: Rocuronium at a dose of 0.45 mg/kg possesses a fairly rapid onset of neuromuscular blockade and has short:intermediate duration of action when used with a desflurane anesthetic. This quality makes it a desirable drug for operations of relatively short duration. Rocuronium at a dose of 0.45 mg/kg has a faster onset and shorter duration than atracurium, at 0.5 mg/kg, when used with a desflurane anesthetic. PMID- 11498312 TI - Small dose of prostaglandin E(1) increases cardiac output without altering blood volume. AB - STUDY OBJECTIVE: To investigate the effect of small dose of intravenous (IV) prostaglandin E(1) (PGE(1)) on blood volume (BV) and cardiac output (CO) by pulse dye-densitometry (PDD) in patients administered isoflurane anesthesia. DESIGN: Prospective, randomized study. SETTING: University hospital. PATIENTS: 14 ASA physical status I and II adult patients undergoing elective neurosurgery. INTERVENTIONS: Patients were randomly assigned to either the PGE(1) group (n = 7) or the control group (n = 7). Anesthesia was induced with thiamylal, fentanyl, and vecuronium, and maintained with isoflurane and nitrous oxide. When the cardiovascular system stabilized after craniotomy and incision of the dura mater, we administered a small dose of PGE(1) at a rate of 0.02 microg/kg/min (PGE(1) group) or saline at a rate of 2 mL/min (control group). MEASUREMENTS AND MAIN RESULTS: Blood volume, CO, and mean transit time (MTT) were measured by PDD before and 60 minutes after the start of administration. At the same timing, mean arterial pressure (MAP), heart rate (HR), and central venous pressure (CVP) were measured, and systemic vascular resistance (SVR), cardiac index (CI), and CO/BV were computed. As for MAP, there was no significant difference within a group and between groups. In the PGE(1) group, significant increases were noted in CI from 2.54 +/- 0.46 to 3.24 +/- 0.83 (mean +/- SD) L/min/m(2) (p < 0.05), in CO/BV from 0.90 +/- 0.24 to 1.19 +/- 0.33 (p < 0.05), and in HR from 65.7 +/- 10.1 to 74.9 +/- 12.1 bpm (p < 0.05), and a significant decrease was observed in MTT from 22.3 +/- 6.5 to 18.2 +/- 5.1 seconds (p < 0.05 ). Cardiac index and CO/BV in the PGE(1) group increased higher than in the control group, while BV, CVP and SVR remained consistent in both groups. CONCLUSIONS: A small dose of PGE(1), low enough not to provoke hypotension, increased CO without alterations in BV. The increase in CO seemed to be mainly due to an increase in HR. PMID- 11498313 TI - Transcranial Doppler ultrasonography with induction of anesthesia and neuromuscular blockade in surgical patients. AB - STUDY OBJECTIVES: To evaluate the cerebral vascular effects of cis-atracurium and rocuronium given after thiopental induction of anesthesia. DESIGN: Randomized, single-blinded study. SETTING: University-affiliated hospital. PATIENTS: 39 adult ASA physical status I and II patients undergoing nonintracranial procedures. INTERVENTIONS: Patients received intravenously (IV), either saline placebo, cis atracurium, or rocuronium after induction of general anesthesia with thiopental sodium. MEASUREMENTS: The right middle cerebral artery was insonated using a pulsed-wave range-gated transcranial Doppler, and data were recorded at preinduction, immediately postinduction, at injection of the study drug, and at 15-second intervals for 3 minutes thereafter. The variables recorded for each subject included the systolic, diastolic, and mean flow velocity, as well as pulsality index, systolic, diastolic, and mean arterial blood pressure (MAP), and end-tidal carbon dioxide concentration. MAIN RESULTS: No significant differences between the groups were present in the postanesthetic induction maximal or minimal mean flow velocity. CONCLUSIONS: cis-Atracurium and rocuronium, administered after thiopental, do not produce clinically relevant changes in cerebral blood flow velocity. PMID- 11498314 TI - Evaluation of the safety and efficacy of epidural ketamine combined with morphine for postoperative analgesia after major upper abdominal surgery. AB - STUDY OBJECTIVE: To evaluate the efficacy of the combination of epidural ketamine and morphine compared with epidural morphine alone for postoperative pain relief following major upper abdominal surgery. STUDY DESIGN: Prospective, randomized, double-blinded study. SETTING: Tertiary care referral and teaching hospital. PATIENTS: 46 ASA physical status I and II patients who underwent major upper abdominal procedures. INTERVENTIONS: Patients were randomly allocated to one of the two treatment groups: patients in Group 1 received epidural morphine 50 microg/kg whereas patients in Group 2 received epidural ketamine 1 mg/kg combined with 50 microg/kg of morphine postoperatively. MEASUREMENTS: A blinded observer using a visual analog scale (VAS) for pain assessment followed up patients for 48 hours postoperatively. Top-up dose of epidural morphine was provided when VAS was higher than 4. Analgesic requirements and side effects were compared between the two groups. RESULTS: Only 40 patients completed the study. There were no differences between the two groups with respect to age, gender, weight, duration, or type of surgical procedure or intraoperative opioid requirements. Onset of analgesia was faster (p < 0.001) in Group 2 (11 min) than in Group 1 patients (25 min). The time for first requirement of analgesia was significantly (p < 0.01) longer (19.8 +/- 9.8 hours) in Group 2 patients than Group 1 (12.8 +/- 6.2 hours). Total number of supplemental doses of epidural morphine required in the first 48 hours postoperatively was also significantly less (p < 0.005) in Group 2 compared to Group 1. Patients in Group 2 had higher sedation scores than Group I patients for the first 2 hours postoperatively. None of the patients in either group developed hallucinations or respiratory depression. Other side effects such as pruritus, nausea, and vomiting were also similar in both groups. CONCLUSIONS: The addition of epidural ketamine 1 mg/kg to morphine 50 microg/kg improved analgesia after major upper abdominal surgery without increasing side effects. PMID- 11498315 TI - The preoperative evaluation form: assessment of quality from one hundred thirty eight institutions and recommendations for a high-quality form. AB - STUDY OBJECTIVE: To develop a rating system to evaluate the preoperative evaluation form in three categories: informational content, ease of use, and ease of reading; to analyze forms contributed by the Association of Anesthesia Clinical Directors; to offer an example of a more optimal form; and interest practitioners in evaluating the quality of their evaluation forms. SETTING: University medical center. DESIGN/MEASUREMENTS: The informational content of the forms was graded by a rating system developed to give a score based on the importance of the data to anesthesia management and legal documentation. Based on the amount of informational content, the forms were grouped into categories ranging from severely deficient to excellent. The ease of use and ease of reading were determined by specific criteria. MAIN RESULTS: 138 forms were analyzed. The mean +/- SD information score was 265 +/- 41, with a range of 140 to 333 (maximum score possible = 363). Fifteen percent of the forms were considered severely deficient to substandard whereas 52% were deemed very good to excellent. A large proportion of forms omitted at least one essential information item. Twelve percent and 6% were labeled poor with regard to ease of use and ease of reading, respectively. Academic centers performed significantly better in the content and ease of use of their forms compared to those from private centers. CONCLUSIONS: A good preoperative evaluation form is an important tool in the anesthetic management of a patient and plays a significant role in the medical legal arena. We found that a surprisingly high percentage of forms are missing important information. A form that contains all of the information and also meets our ease of use and reading standards was developed and offered as an example. PMID- 11498316 TI - Variation in practice patterns of anesthesiologists in California for prophylaxis of postoperative nausea and vomiting. AB - STUDY OBJECTIVE: To assess the responses to a survey asking anesthesiologists to report their clinical practice patterns for postoperative nausea and vomiting (PONV) prophylaxis. These practice patterns data may be useful for understanding how to optimize the decision to provide PONV prophylaxis. DESIGN: A written questionnaire with three detailed clinical scenarios with differing levels of a priori risk of PONV (a low-risk patient, a medium-risk patient, and a high-risk patient) was mailed to 454 anesthesiologists. SETTING: Survey was completed by anesthesiologists (n = 240) in 3 university and 3 community practices in California. MEASUREMENTS: Type and number of pharmacological and nonpharmacological interventions for PONV prophylaxis were recorded. To assess the variability in the responses (by the a priori risk of patient), we counted the number of different regimens that would be necessary to account for 80% of the responses. MAIN RESULTS: For the 240 respondents, we found that 1, 9, and 11 different pharmacological prophylaxis regimens were required to account for 80% of the variability in practice patterns for the low-, medium-, and high-risk patients, respectively. For the low-risk patient, 19% of practitioners would use pharmacological prophylaxis, and 37% would use nonpharmacological prophylaxis. For the medium-risk patient, 61% would use nonpharmacological prophylaxis and 67% of practitioners would use multidrug prophylaxis: 45% of patients would receive a 5HT(3) antagonist, 35% would receive metoclopramide, and 16% would receive droperidol. For the high-risk patient, 94% of practitioners would administer a 5HT(3) antagonist, whereas 84% would use multi-drug prophylaxis. CONCLUSIONS: We found a wide range of PONV prophylaxis management patterns. This variation in clinical practice may reflect uncertainty about the efficacy of available interventions, or differences in practitioners' clinical judgment and beliefs about how to treat PONV. Some therapies with proven benefit for PONV may be underused. Our results may be useful for designing studies aimed at determining the impact on PONV rates when physicians develop and implement guidelines for PONV prophylaxis. PMID- 11498317 TI - Influence of pneumoperitoneum and patient positioning on respiratory system compliance. AB - STUDY OBJECTIVE: To investigate the influence of pneumoperitoneum (PP) and posture on respiratory compliance and ventilation pressures. DESIGN: Prospective, single blind trial. PATIENTS: 10 female ASA physical status I and II patients scheduled for elective gynecologic laparoscopy. SETTING: University medical center. INTERVENTIONS: Anesthesia was performed as total IV anesthesia (TIVA) with propofol, alfentanil, and atracurium. After induction of anesthesia and orotracheal intubation, the lungs were ventilated to maintain partial pressure of CO(2) (P(ET)CO(2)) of 30 +/- 3 mmHg. Ventilation was kept constant. As gas mixture oxygen and air 1:1 was used without positive end-expiratory pressure (PEEP). MEASUREMENTS: Measurements were taken before and after creation of pneumoperitoneum with an intraabdominal pressure (IAP) of 10 mmHg, of 15 mmHg in 20 degrees head-down tilt, then in 20 degrees head-up tilt, and after deflation of PP. We determined peak inspiratory pressure (PIP), mean airway pressure (mPaw), P(ET)CO(2), expiratory minute volume (V(E)), heart rate (HR), and systolic (SBP), diastolic (DBP), and mean arterial pressure (MAP). Respiratory system compliance (C(eff rs)) was calculated as quotient of tidal volume (V(T)) and PIP. MAIN RESULTS: After creation of PP (IAP 10 mmHg), there was a significant increase of median PIP (3 cmH(2)O), mPaw (1 cm H(2)O) and arterial pressure (BP), (MAP by 7 mmHg), C(eff rs) decreased by 6 mL. cm H(2)O(-1). Increase of IAP to 15 mmHg led to a further increase of PIP (2 cm H(2)O) and mPaw (1 cm H(2)O), and a further decrease of C(eff rs) by 5 mL cm H(2)O(-1); BP decreased (MAP by 5.5 mmHg). Head-up or head down positions showed no significant hemodynamic or pulmonary changes. P(ET)CO(2)increased from 29.5 to 36 mmHg at an IAP of 15 mmHg, but then no further changes were noticed. Five minutes after deflation of pneumoperitoneum all values returned to baseline levels. CONCLUSIONS: Creation of PP at an IAP of 15 mmHg reduced respiratory system compliance, and increased peak inspiratory and mean airway pressures, which quickly returned to normal values after deflation. Head-down or head-up position did not further alter those parameters. PMID- 11498318 TI - Dose of prophylactic intravenous ephedrine during spinal anesthesia for cesarean section. AB - STUDY OBJECTIVE: To compare the incidence of maternal hypotension associated with spinal anesthesia for cesarean section when 10-, 15-, or 20-mg prophylactic boluses of intravenous (IV) ephedrine are used. DESIGN: Prospective observational study. SETTING: Teaching hospital. PATIENTS: 108 women admitted for elective cesarean section. INTERVENTIONS: Spinal anesthesia was performed using hyperbaric bupivacaine 10 mg, sufentanil 2 microg, and morphine 0.2 mg (volume 4 mL). Ephedrine (10, 15, or 20 mg) was administered 2 minutes after the intrathecal injection. Maternal blood pressure was checked every 2 minutes. Hypotension was promptly treated with 5-mg ephedrine boluses. MAIN RESULTS: Incidence of hypotension was significantly higher in women receiving a 10-mg prophylactic dose of ephedrine than in those receiving either a 15-mg or a 20-mg prophylactic dose of ephedrine [23/36 in the 10-mg ephedrine group vs. 13/36 and 10/36 in the 15-mg and 20-mg ephedrine groups, respectively (p< 0.05)]. CONCLUSION: In the conditions of this study, a single bolus of IV ephedrine with doses of either 15 or 20 mg decreased significantly the incidence of maternal hypotension as compared to a single 10-mg bolus of ephedrine. PMID- 11498319 TI - A study of 13 patients with gastric tube in place after esophageal resection: use of omeprazole to decrease gastric acidity and volume. AB - STUDY OBJECTIVE: To investigate whether oral omeprazole 20 mg decreases the risk of aspiration pneumonia in patients with gastric tube reconstruction. DESIGN: Consecutive study. SETTING: Operation room of cancer center. PATIENTS: Thirteen patients with gastric tube reconstruction for esophageal cancer. INTERVENTIONS: Oral omeprazole 20 mg was given the night before surgery. A rapid-sequence induction with cricoid pressure was employed for induction of anesthesia. After tracheal intubation, a nasogastric catheter was inserted into the gastric tube and the contents were aspirated. MEASUREMENTS AND MAIN RESULTS: The pH and volume of the gastric contents were measured. The pH and volume of the gastric tube contents were 4.5 +/- 1.6 (range from 2.5 to 7.0) and 9.5 +/- 10.2 mL (range from 0 to 30 mL), respectively. Food residue was recognized in nine patients. There was no patient with a pH below 2.5 and a volume of 25 mL or greater. CONCLUSIONS: Omeprazole 20 mg decreased the acidity and volume of the gastric tube contents and reduced the risk of aspiration pneumonia in patients with a gastric tube in place. PMID- 11498320 TI - Anesthesiologists' interest in neonatal resuscitation certification. AB - STUDY OBJECTIVE: To survey graduates from our residency program to determine their involvement and their interest in becoming certified in neonatal resuscitation. DESIGN: Survey questionnaire. SETTING: Graduates from the University of Pennsylvania Anesthesia Program between 1989 and 1999. SUBJECTS: 212 individuals completed the residency; addresses were available for 189 and the response rate for these individuals was 156. RESULTS: Of the respondents with obstetric anesthesia responsibilities, 65% have been involved in resuscitation of the newborn. Sixteen percent were certified, but 81% were interested in certification. If the hospital had less than 1000 deliveries per year, the probability of the anesthesiologist having to perform neonatal resuscitation was 11.0 times greater than if the hospital had more than 1000 deliveries per year (confidence interval, 1.4--86). CONCLUSIONS: Despite guidelines recommending the contrary, anesthesiologists are involved in the resuscitation of the newborn. The majority of these individuals are not certified but are interested in becoming certified. The ASA should establish programs to offer the opportunity to learn and to reinforce these skills. PMID- 11498321 TI - Incidence and indications for reintubation during postoperative care following orthotopic liver transplantation. AB - STUDY OBJECTIVE: To analyze the incidence and indications for reintubation during postoperative care following orthotopic liver transplantation (OLT). DESIGN: Retrospective chart review. SETTING: Large metropolitan teaching hospital. PATIENTS: 546 adult liver transplant recipients. MEASUREMENTS AND MAIN RESULTS: The medical charts of 546 patients who underwent OLT at our institution between January 1992 and September 1996 were reviewed for the incidence and indications of reintubation throughout primary hospitalization. Eighty-one of 546 patients (14.8%) required one or more episodes of reintubation after OLT. In the majority of cases, reintubation was performed for pulmonary complications (44.6%), followed by cerebral (19.1%) and surgical (14.5%) complications. Cardiac (9.1%) and peripheral neurologic (2.7%) complications were less frequent reasons for reintubation. Overall patient survival, according to the Kaplan-Meier estimates, was 89.9%, 87.5%, 86.5%, and 82.2% after 1, 2, 3, and 5 years, respectively. In patients with one or more episodes of reintubation, overall survival decreased to 62.5% after 1, 2, and 3 years, and to 56.4% after 5 years (p < 0.001). CONCLUSIONS: The main indications for reintubation after OLT were pulmonary, cerebral, and surgical complications. These reintubation events had a considerable influence on the patient's postoperative recovery, and were associated with a significantly higher rate of mortality, than for OLT patients who did not undo reintubation. PMID- 11498322 TI - Asymptomatic vallecular cyst: airway management considerations. AB - Airway problems are easiest to manage when they are anticipated. Difficult intubation might, however, occur in patients with no obvious signs or symptoms suggesting airway difficulty. We describe a case where laryngeal inlet was obscured by a large vallecular cyst that was discovered during rapid-sequence induction of general anesthesia, causing difficulty in tracheal intubation. Once the patient was allowed to recover from general anesthesia, the trachea could be safely intubated using a fiberoptic bronchoscope. PMID- 11498323 TI - Anesthetic management of the exit (ex utero intrapartum treatment) procedure. AB - The EXIT (ex utero intrapartum treatment) procedure is used to maintain fetal placental circulation during partial delivery of a fetus with a potentially life threatening upper airway obstruction. We performed the EXIT procedure on a fetus with a large intra-oral cyst. Sevoflurane was used as the anesthetic because of its rapid titratability. Sevoflurane provided excellent maternal and fetal anesthesia. Modifications to previously described monitoring techniques for the EXIT procedure were also used. PMID- 11498324 TI - Subdural air collection: a likely source of radicular pain after lumbar epidural. AB - This case conference reports two cases of epidural anesthesia in which air was used to identify the epidural space during a loss-of-resistance placement technique. Both patients subsequently complained of severe pain and subdural air was demonstrated in case 1 by computed tomography and in case 2 by magnetic resonance imaging. The possible causes of the pain syndrome experienced by both patients are discussed. PMID- 11498325 TI - Leaf area distribution and radiative transfer in open-canopy forests: implications for mass and energy exchange. AB - Leaf area and its spatial distribution are key canopy parameters needed to model the radiation regime within a forest and to compute the mass and energy exchange between a forest and the atmosphere. A much larger proportion of available net radiation is received at the forest floor in open-canopy forests than in closed canopy forests. The proportion of ecosystem water vapor exchange (lambda E) and sensible heat exchange from the forest floor is therefore expected to be larger in open-canopy forests than in closed-canopy forests. We used a combination of optical and canopy geometry measurements, and robust one- and three-dimensional models to evaluate the influence of canopy architecture and radiative transfer on estimates of carbon, water and energy exchange of a ponderosa pine (Pinus ponderosa Dougl. ex Laws.) forest. Three-dimensional model simulations showed that the average probability of diffuse and direct radiation transmittance to the forest floor was greater than if a random distribution of foliage had been assumed. Direct and diffuse radiation transmittance to the forest floor was 28 and 39%, respectively, in the three-dimensional model simulations versus 23 and 31%, respectively, in the one-dimensional model simulations. The assumption of randomly distributed foliage versus inclusion of clumping factors in a one dimensional, multi-layer biosphere-atmosphere gas exchange model (CANVEG) had the greatest effect on simulated annual net ecosystem exchange (NEE) and soil evaporation. Assuming random distribution, NEE was 41% lower, net photosynthesis 3% lower, total lambda E 10% lower, and soil evaporation 40% lower. The same comparisons at LAI 5 showed a similar effect on annual NEE estimates (37%) and lambda E (12%), but a much larger effect on net photosynthesis (20%), suggesting that, at low LAI, canopies are mostly sunlit, so that redistribution of light has little effect on net photosynthesis, whereas the effect on net photosynthesis is much greater at high LAIs. PMID- 11498326 TI - A theoretical analysis of the influence of heterogeneity in chlorophyll distribution on leaf reflectance. AB - Attempts to determine the vitality of vegetation and to detect vegetation stress from remotely sensed data have focused on chlorophyll concentration, because it influences the reflectance of vegetation and tends to correlate with vegetation health and stress. Pollution, pathogens and pests can cause localized regions of chlorosis and necrosis across a leaf surface, but the extent to which these patches influence the overall reflectance and spectral signature of the leaf and canopy has not been tested. A conifer leaf model (LIBERTY), which simulates the influence of leaf biochemical concentrations of chlorophyll, water, lignin, cellulose and protein on the reflectance of leaves from 400 to 2500 nm, was used to determine the effect of patches of chlorosis on leaf reflectance. A fraction of the leaf f is assumed to be chlorotic with a chlorophyll concentration C(1). The remainder of the leaf has chlorophyll concentration C(2) such that mean leaf chlorophyll concentration, C(mean) = fC(1) + (1 - f)C(2), is constant for a range of f and C(1) values. LIBERTY can be used to estimate the reflectance of a leaf with a particular chlorophyll concentration at a particular wavelength R(lambda,C) (assuming other leaf properties remain constant), thus we can estimate the reflectance of the chlorotic leaf as fR(lambda,C(1))+ (1 - f)R(lambda,C(2)). The model indicated that small areas of chlorosis have a disproportionately large influence on overall leaf reflectance. For example, a leaf with 25% of its area chlorotic can have the same reflectance (400-700 nm) as a homogeneous leaf with 60% less chlorophyll. Thus, determination of chlorophyll concentration from remotely sensed data is prone to underestimation when chlorophyll is nonuniformly distributed. Hence, attempts to model leaf and canopy reflectance using radiative transfer models will need to consider how to incorporate nonuniform chlorophyll distribution. PMID- 11498327 TI - A method for estimating light interception by a conifer shoot. AB - We present an operational method for estimating the amount of PAR intercepted by a coniferous shoot. Interception of PAR by a shoot is divided into three components: the amount of radiation coming from the sky, the transmission of radiation through the surrounding vegetation, and the shoot' s silhouette area facing the direction of the incoming radiation. All three components usually vary with direction. Radiation incident from the sky consists of direct and diffuse radiation. The well-known equation of motion for the sun and Beer' s Law for atmospheric transmittance are used to simulate the directional distribution of direct sunlight for any given period of time. The diffuse component is assumed to be uniform. Meteorological field measurements are used to calibrate the absolute amounts of the direct and diffuse components. The gap fraction (proportion of visible sky) in different directions around a shoot is measured by analyzing a hemispherical fish-eye photograph, taken at the location of the shoot, with an image processing program. Similarly, the shoot silhouette area (SSA) is measured by photographing the shoot from many different directions. The measurements of SSA are interpolated by a method called trigonometric interpolation to obtain the directional distribution of SSA over the entire hemisphere. This distribution is then rotated according to the shoot' s position in the canopy. Multiplying incoming PAR, canopy gap fraction and SSA in different directions, and summing over all directions, gives an estimate of PAR intercepted by the shoot during the chosen period of time. The method is described step by step, and applied, as an example, to a shoot from a Scots pine (Pinus sylvestris L.) stand in central Finland. Differences in radiation interception properties between sun and shade shoots and their relevance to canopy-scale models are discussed. PMID- 11498328 TI - Shoot structure and photosynthetic efficiency along the light gradient in a Scots pine canopy. AB - We examined the effects of structural and physiological acclimation on the photosynthetic efficiency of Scots pine (Pinus sylvestris L.) shoots. We estimated daily light interception (DLI) and photosynthesis (DPHOT) of a number of sample shoots situated at different positions in the canopy. Photosynthetic efficiency (epsilon) was defined as the ratio of DPHOT to the potential daily light interception (DLI(ref)) defined as the photosynthetically active radiation (PAR) intercepted per unit area of a sphere at the shoot location. To calculate DLI(ref), DLI and DPHOT, the radiation field surrounding a shoot in the canopy was first modeled using simulated directional distributions of incoming PAR on a clear and an overcast day, and estimates of canopy gap fraction in different directions provided by hemispherical photographs. A model of shoot geometry and measured data on shoot structure and photosynthetic parameters were used to simulate the distribution of PAR irradiance on the needle surface area of the shoot. Photosynthetic efficiency (epsilon) was separated into light-interception efficiency (epsilon(I) = DLI/DLI(ref)) and conversion efficiency (epsilon(PHOT) = DPHOT/DLI). This allowed us to quantify separately the effect of structural acclimation on the efficiency of photosynthetic light capture (epsilon(l)), and the effect of physiological acclimation on conversion efficiency (epsilon(PHOT)). The value of epsilon increased from the top to the bottom of the canopy. The increase was largely explained by structural acclimation (higher epsilon(I)) of the shade shoots. The value of epsilon(PHOT) of shade foliage was similar to that of sun foliage. Given these efficiencies, the clear-day value of DPHOT for a sun shoot transferred to shade was only half that of a shade shoot at its original position. The method presented here provides a tool for quantitatively estimating the role of acclimation in total canopy photosynthesis. PMID- 11498329 TI - Importance of needle age and shoot structure on canopy net photosynthesis of balsam fir (Abies balsamea): a spatially inexplicit modeling analysis. AB - We have developed a spatially inexplicit model of canopy photosynthesis for balsam fir (Abies balsamea (L.) Mill.) that accounts for key processes of light shoot interaction including irradiance interception by the shoot, spatial aggregation of shoots into branches and crowns, the differential propagation of diffuse and direct light within the canopy, and an ideal representation of penumbra. Also accounted for in the model are the effects of the average radiative climate and shoot age on needle retention, light interception, and photosynthetic capacity. We used reduced versions of this model to quantify the effects of simplifying canopy representation on modeled canopy net photosynthesis. Simplifications explored were the omission of direct beam transformation into penumbral light and the use of different constant shoot properties throughout the canopy. The model was parameterized for a relatively dense balsam fir stand (leaf area index of 5.8) north of Quebec City, Canada, and run using hourly meteorological data obtained at the site. The overall performance of the complete model was satisfactory, with maximum values of canopy net photosynthesis of 23 micromol (m(2) ground)(-1) s(-1) (83 mmol m(-2) h(-1)), and a near-saturation of the canopy at a photosynthetically active radiation photon flux density of about 750 micromol m(-2) s(-1) (2.7 mol m(-2) h(-1)). The omission of penumbral effects through the use of unattenuated direct (beam) radiation at all layers of the canopy, as used for broad-leaved species, reduced canopy net photosynthesis by 3.7%. Analysis of the results show that the small impact of penumbra on canopy net photosynthesis stems from the high proportion of diffuse radiation (73%) estimated from our meteorological data set; single-hour results under clear sky conditions approach theoretical bias values of about 30%. Use of mean shoot photosynthetic, light capture and light transmission properties throughout the canopy biased canopy net photosynthesis by less than 3%. However, simulations carried out based on properties of 1-year-old shoots throughout the canopy overestimated canopy net photosynthesis by 9%. Use of the shoot as our smallest functional unit was a potential source of bias because the differential absorption of direct and diffuse radiation within the shoot could not be factored into the model. Other sources of potential bias are discussed. PMID- 11498330 TI - Increased understanding of nutrient immobilization in soil organic matter is critical for predicting the carbon sink strength of forest ecosystems over the next 100 years. AB - The terrestrial biosphere is currently thought to be a significant sink for atmospheric carbon (C). However, the future course of this sink under rising [CO2] and temperature is uncertain. Some contrasting possibilities that have been suggested are: that the sink is currently increasing through CO2 fertilization of plant growth but will decline over the next few decades because of CO2 saturation and soil nutrient constraints; that the sink will continue to increase over the next century because rising temperature will stimulate the release of plant available soil nitrogen (N) through increased soil decomposition; that, alternatively, the sink will not be sustained because the additional soil N released will be immobilized in the soil rather than taken up by plants; or that the sink will soon become negative because loss of soil C through temperature stimulation of soil respiration will override any CO2 or temperature stimulation of plant growth. Soil N immobilization is thus a key process; however, it remains poorly understood. In this paper we use a forest ecosystem model of plant-soil C and N dynamics to gauge the importance of this uncertainty for predictions of the future C sink of forests under rising [CO2] and temperature. We characterize soil N immobilization by the degree of variability of soil N:C ratios assumed in the model. We show that the modeled C sink of a stand of Norway spruce (Picea abies (L.) Karst.) in northern Sweden is highly sensitive to this assumption. Under increasing temperature, the model predicts a strong C sink when soil N:C is inflexible, but a greatly reduced C sink when soil N:C is allowed to vary. In complete contrast, increasing atmospheric [CO2] leads to a much stronger C sink when soil N:C is variable. When both temperature and [CO2] increase, the C sink strength is relatively insensitive to variability in soil N:C; significantly, however, with inflexible soil N:C the C sink is primarily a temperature response whereas with variable soil N:C, it is a combined temperature-CO2 response. Simulations with gradual increases of temperature and [CO2] indicate a sustained C sink over the next 100 years, in contrast to recent claims that the C sink will decline over the next few decades. Nevertheless, in using a relatively simple model, our primary aim is not to make precise predictions of the C sink over the next 100 years, but rather to highlight key areas of model uncertainty requiring further experimental clarification. Here we show that improved understanding of the processes underlying soil N immobilization is essential if we are to predict the future course of the forest carbon sink. PMID- 11498331 TI - Mean canopy stomatal conductance responses to water and nutrient availabilities in Picea abies and Pinus taeda. AB - We compared sap-flux-scaled, mean, canopy stomatal conductance (GS) between Picea abies (L.) Karst. in Sweden and Pinus taeda (L.) in North Carolina, both growing on nutritionally poor soils. Stomatal conductance of Picea abies was approximately half that of Pinus taeda and the sensitivity of GS in Picea abies to vapor pressure deficit (D) was lower than in Pinus taeda. Optimal fertilization increased leaf area index (L) two- and threefold in Pinus taeda and Picea abies, respectively, regardless of whether irrigation was increased. Although it increased L, fertilization did not increase GS in Picea abies unless irrigation was also provided. In Pinus taeda growing on coarse, sandy soils, the doubling of L in response to fertilization reduced GS sharply unless irrigation was also provided. The reduction in GS with fertilization in the absence of irrigation resulted from the production of fine roots with low saturated hydraulic conductivity. When Pinus taeda received both fertilization and irrigation, the increase in L was accompanied by a large increase in GS. In Pinus taeda, a reference GS (defined as GS at D = 1 kPa; GSR) decreased in all treatments with decreasing volumetric soil water content (theta). In Picea abies, theta varied little within a treatment, but overall, GSR declined with theta, reaching lowest values when drought was imposed by the interception of precipitation. Despite the large difference in GS both between Picea abies and Pinus taeda and among treatments, stem growth was related to absorbed radiation, and stem growth response to treatment reflected mostly the changes in L. PMID- 11498332 TI - Effects of nutrition and soil water availability on water use in a Norway spruce stand. AB - We investigated effects of nutrition and soil water availability on sap flux density, transpiration per unit leaf area (EL), and canopy stomatal conductance (GS) of Norway spruce (Picea abies L. (Karst.)) in northern Sweden during the 1996 growing season. Our objectives were to determine (1) if artificially imposed drought (65% rain diversion) reduces soil water sufficiently to cause physiological limitations to whole-tree and plot-scale water transport, and (2) whether increased capacity for water transport resulting from fertilization induced increases in leaf (> 3-fold) and sapwood areas (> 2.3-fold) deplete soil water sufficiently to cause a negative feedback on GS and EL. We monitored soil water content (theta) and soil water potential (PsiS) in control (C), drought (D), fertilized (F) and irrigated + fertilized (IL) treatment plots, along with site meteorological conditions. Ten trees per plot were monitored for sap flow. Although there were significant treatment differences in mean daily EL (C > D > F; P < 0.01) and GS (C > D > F; P < 0.05), variation in absolute magnitudes was small. Therefore, transpiration differences on a unit ground area basis (EC) were nearly proportional to leaf area differences. Precipitation was well distributed throughout the study period and so PsiS remained high, except during short dry periods in Plot F when it declined rapidly. Thus, although soil water was not limiting to GS, EL or EC when precipitation was uniformly distributed throughout the growing season, we cannot conclude that water availability would not limit GS in fertilized stands if the seasonal distribution of precipitation were altered. PMID- 11498333 TI - Seasonal and interannual variations in carbon isotope discrimination in a maritime pine (Pinus pinaster) stand assessed from the isotopic composition of cellulose in annual rings. AB - Stable carbon isotope composition (delta; per thousand) was measured on cellulose extracted from maritime pine (Pinus pinaster Ait.) tree rings to investigate inter-tree and interannual variability (7 trees, 20 rings per tree, each ring divided into early and late wood). A model of stand primary production coupled to water balance was used to calculate the stand annual water-use efficiency (WUE). Inter-tree variability in discrimination (maximum 2.88 per thousand in late wood in 1989, 2.69 per thousand in early wood in 1983) was as large as interannual variation (maximum 2.72 per thousand in late wood, 2.05 per thousand in early wood). Tree size did not explain these differences. Relationships were found between annual discrimination and climate variables such as annual rainfall, summer temperature and vapor pressure deficit (VPD). Higher correlations were found with late wood discrimination. Early wood discrimination was shown to be related to previous-year late wood discrimination. Late wood discrimination was also related to soil water availability. Stand annual WUE was only weakly related to tree ring carbon discrimination. PMID- 11498334 TI - Link between diurnal stem radius changes and tree water relations. AB - Internal water reserves are depleted and replenished daily, not only in succulent plants, but also in trees. The significance of these changes in tissue water storage for tree water relations was investigated by monitoring diurnal fluctuations in stem radius. In 6-year-old potted Norway spruce (Picea abies (L.) Karst.) trees, whole-tree transpiration rate (T), sap flow at the stem base and fluctuations in stem radius were measured at 10-min intervals over eight successive weeks. The dynamics of diurnal water storage in relation to the daily course of water movement was simulated and the contribution of stored water to T quantified. The finding that, in P. abies, the course of bark water content is linearly coupled to stem radius fluctuations provided the basis for linking stem radius changes to a functional flow and storage model for tree water relations. This model, which consists of physical functions only and is driven by a single input variable (T), accurately simulates the diurnal course of changes in stem radius and water storage of the tree crown and stem. It was concluded that fluctuations were mainly determined by the course of transpiration. The availability of soil water and the degree to which storage tissues were saturated were also factors affecting the diurnal course of stem radius changes. Internally stored water contributed to daily transpiration even in well-watered trees, indicating that stored water plays an important role not only during periods of drought, but whenever water transport occurs within the tree. Needle and bark water reserves were most heavily depleted during transpiration. Together they supplied approximately 10% of daily T on sunny days, and up to 65% on cloudy days. On a daily basis, the crown (mainly needles) contributed approximately eight times more water to T than the stem (mainly bark). The depletion of the two storage pools and the water movements observed in the trees always occurred in the same sequence. In the morning, T first caused a depletion of the water stored in the crown. It then caused depletion of bark storage tissues at ever increasing distances from the needles. Up to 75% of the transpired water could be withdrawn from storage tissues when the increase in T reached a maximum. PMID- 11498335 TI - Hydraulic and stomatal adjustment of Norway spruce trees to environmental stress. AB - A study of how the water conducting systems of 30-50-year-old Norway spruce (Picea abies (L.) Karst.) trees growing at three sites adjust to shade and waterlogging indicated that water relations characteristics varied with the life histories of the trees. Xylem was more efficient at conducting water and stomata were more sensitive to atmospheric evaporative demand in trees subjected to favorable growth conditions (control trees) than in trees growing in shade or waterlogged conditions. At the same soil water availability, shade-grown trees suffered more severely from water deficit than control trees. Under conditions of high atmospheric vapor pressure deficit, foliage of shade-grown trees exhibited low water potentials, as a result of low hydraulic conductance of the vascular system and inefficient stomatal control. Because of the increased internal resistance to water flow, more negative leaf water potentials (Psi(x)) must be reached to provide an adequate water supply to the foliage. It is concluded that dynamic water stress is one of the main causes of the continuing growth retardation in suppressed Norway spruce trees after their release from the overstory. Trees growing in waterlogged soil (bog-grown trees) were characterized by weak stomatal control, resulting in large water losses from the foliage. Although bog-grown trees exhibited uneconomical water use, they possessed mechanisms (e.g., osmotic adjustment) that allowed leaves to tolerate low Psi(x) while stomata remained open. Under conditions of sufficient soil water availability and moderate atmospheric vapor pressure deficit, soil-to-leaf conductance was highest in bog-grown trees (1.45 +/- 0.06 mmol m(-2) s(-1) MPa( 1)), followed by control and shade-grown trees (1.04 +/- 0.04 and 0.77 +/- 0.05 mmol m(-2) s(-1) MPa(-1), respectively). The lowest soil-to-leaf conductance (0.45 +/- 0.04 mmol m(-2) s(-1) MPa(-1)) was recorded in control trees at high atmospheric evaporative demand, and was probably caused by tracheid cavitation. PMID- 11498336 TI - Tree stem diameter variations and transpiration in Scots pine: an analysis using a dynamic sap flow model. AB - A dynamic model for simulating water flow in a Scots pine (Pinus sylvestris L.) tree was developed. The model is based on the cohesion theory and the assumption that fluctuating water tension driven by transpiration, together with the elasticity of wood tissue, causes variations in the diameter of a tree stem and branches. The change in xylem diameter can be linked to water tension in accordance with Hookea s law. The model was tested against field measurements of the diurnal xylem diameter change at different heights in a 37-year-old Scots pine at Hyytiala, southern Finland (61 degrees 51' N, 24 degrees 17' E, 181 m a.s.l.). Shoot transpiration and soil water potential were input data for the model. The biomechanical and hydraulic properties of wood and fine root hydraulic conductance were estimated from simulated and measured stem diameter changes during the course of 1 day. The estimated parameters attained values similar to literature values. The ratios of estimated parameters to literature values ranged from 0.5 to 0.9. The model predictions (stem diameters at several heights) were in close agreement with the measurements for a period of 6 days. The time lag between changes in transpiration rate and in sap flow rate at the base of the tree was about half an hour. The analysis showed that 40% of the resistance between the soil and the top of the tree was located in the rhizosphere. Modeling the water tension gradient and consequent woody diameter changes offer a convenient means of studying the link between wood hydraulic conductivity and control of transpiration. PMID- 11498337 TI - Sensitivity of photosynthetic electron transport to photoinhibition in a temperate deciduous forest canopy: Photosystem II center openness, non-radiative energy dissipation and excess irradiance under field conditions. AB - We used chlorophyll fluorescence techniques to investigate responses of Photosystem II (PSII) quantum yield to light availability in the short term (quantum flux density integrated over the measurement day, Qd) and in the long term (Qd averaged over the season, Qs) in a mixed deciduous forest comprising shade-tolerant and water-stress-sensitive Tilia cordata Mill. in the lower canopy and shade-intolerant and water-stress-resistant Populus tremula L. in the upper canopy. In both species, intrinsic efficiency of PSII in the dark-adapted state (Fv/Fm) was lower during the day than during the night, and the difference in Fv/Fm between day and night increased with increasing Qs. Although the capacity for photosynthetic electron transport increased with increasing Qs in both species, maximum quantum efficiency of PSII in the light-adapted state (alpha) decreased with increasing Qs. At a common Qs, alpha was lower in T. cordata than in P. tremula primarily because of a higher fraction of closed PSII centers, and to a smaller extent because of limited, non-radiative, excitation energy dissipation in the pigment bed in T. cordata. Across both species, photochemical quenching (qP), which measures the openness of PSII centers, varied more than fivefold, but the efficiency of excitation energy capture by open PSII centers (Fv'/Fm'), which is an estimate of non-radiative excitation energy dissipation in PSII antennae, varied by only 50%. Chlorophyll turnover rates increased with increasing irradiance, especially in T. cordata, possibly because of increased photodestruction. Diurnal measurements of PSII quantum yields (PhiPSII) indicated that, under similar environmental conditions, PhiPSII was always lower in the afternoon than in the morning, and the fraction of daily integrated photosynthetic electron transport lost because of diurnal declines in PhiPSII (Delta) increased with increasing Qd. At a common Qd, mean daily PSII center reduction state, the fraction of light in excess (1 - fractions of light used in photochemistry and dissipated as heat) and Delta were higher in T. cordata than in P. tremula. This was attributed to greater stomatal closure during the day, which led to a greater reduction in the requirement for assimilative electron flow in T. cordata. Across both species, Delta scaled negatively with the fraction of light utilized photochemically, demonstrating the leading role of PSII center openness in maintaining high PSII efficiency. Because photosynthesis (A) at current ambient carbon dioxide concentration is limited by CO2 availability in high light and mainly by photosynthetic electron transport rates in low light, overall daily down-regulation of PhiPSII primarily influences A in low light. Given that foliar water stress scales positively with Qs in both species, we conclude that the inverse patterns of variation in water and light availabilities in the canopy result in a greater decline in A than is predicted by decreases in stomatal conductance alone. PMID- 11498338 TI - Canopy position and needle age affect photosynthetic response in field-grown Pinus radiata after five years of exposure to elevated carbon dioxide partial pressure. AB - Photosynthesis of tree seedlings is generally enhanced during short-term exposure to elevated atmospheric CO2 partial pressure, but longer-term studies often indicate some degree of photosynthetic adjustment. We present physiological and biochemical evidence to explain observed long-term photosynthetic responses to elevated CO2 partial pressure as influenced by needle age and canopy position. We grew Pinus radiata D. Don. trees in open-top chambers for 5 years in sandy soil at ambient (36 Pa) and elevated (65 Pa) CO2 partial pressures. The trees were well watered and exposed to natural light and ambient temperature. In the fourth year of CO2 exposure (fall 1997), when foliage growth had ceased for the year, photosynthetic down-regulation was observed in 1-year-old needles, but not in current-year needles, suggesting a reduction in carbohydrate sink strength as a result of increasing needle age (Turnbull et al. 1998). In 5-year-old trees (spring 1997), when foliage expansion was occurring, photosynthetic down regulation was not observed, reflecting significantly large sinks for carbohydrates throughout the tree. Net photosynthesis was stimulated by 79% in trees growing in elevated CO2 partial pressure, but there was no significant effect on photosynthetic capacity or Rubisco activity and concentration. Current year needles were more responsive to elevated CO2 partial pressure than 1-year old needles, exhibiting larger relative increases in net photosynthesis to elevated CO2 partial pressure (98 versus 64%). Lower canopy and upper canopy leaves exhibited similar relative responses to growth in elevated CO2 partial pressure. However, needles in the upper canopy exhibited higher net photosynthesis, photosynthetic capacity, and Rubisco activity and concentration than needles in the lower canopy. Given that the ratio of mature to juvenile foliage mass in the canopy will increase as trees mature, we suggest that trees may become less responsive to elevated CO2 partial pressure with increasing age. We conclude that tree response to elevated CO2 partial pressure is based primarily on sink strength and not on the duration of exposure. PMID- 11498339 TI - Photosynthesis and light-use efficiency by plants in a Canadian boreal forest ecosystem. AB - Measurements of the photosynthetic response to midsummer irradiance were made for 11 species representing the dominant trees, understory shrubs, herbaceous plants and moss species in an old black spruce (Picea mariana (Mill.) B.S.P.) boreal forest ecosystem. Maximum rates of photosynthesis per unit foliage area at saturating irradiance, A(max), were highest for aspen (Populus tremuloides Michx.), reaching 16 micromol m(-2) s(-1). For tamarack (Larix laricina (Du Roi) K. Kock) and P. mariana, Amax was only 2.6 and 1.8 micromol m(-2) s(-1), respectively. Values of A(max) for understory shrubs and herbaceous plants were clustered between 9 and 11 micromol m(-2) s(-1), whereas A(max) of feather moss (Pleurozium schreberi (Brid.) Mitt.) reached only 1.9 micromol m(-2) s(-1). No corrections were made for differences in shoot structure, but values of photosynthetic light-use efficiency were similar for most species (70-80 mmol CO2 mol(-1)); however, they were much lower for L. laricina and P. mariana (15 mmol CO2 mol(-1)) and much higher for P. schreberi (102 m;mol CO2 mol(-1)). There was a linear relationship between Amax and foliage nitrogen concentration on an area basis for the broad-leaved species in the canopy and understory, but the data for P. mariana, L. laricina and P. schreberi fell well below this line. We conclude that it is not possible to scale photosynthesis from leaves to the canopy in this ecosystem based on a single relationship between photosynthetic rate and foliage nitrogen concentration. PMID- 11498340 TI - Atmospheric carbon dioxide concentration, nitrogen availability, temperature and the photosynthetic capacity of current-year Norway spruce shoots. AB - Branches of field-grown Norway spruce (Picea abies (L.) Karst.) trees were exposed to either long-term ambient or to elevated CO2 concentrations ([CO2]) using the branch bag technique. The light-saturated photosynthetic rates (A(max)) of current-year shoots differing in nitrogen (N) status were measured at various temperatures and at either ambient (360 micromol mol(-1), AMB) or elevated (ambient + 350 micromol mol(-1), EL) [CO2]. The value of A(max) was determined at various intercellular [CO2]s (A/Ci curves) and used to normalize photosynthetic rates to the mean treatment C(i) values, which were 200 micromol mol(-1) (AMB) and 450 micromol mol(-1) (EL), respectively. Needle N status and temperature strongly affected A(max). The response to N increased with temperature, and the photosynthetic temperature optimum increased with N status. This was assumed to be a result of reduced mesophyll CO2 conductance. The relative increase of Amax in the EL treatment compared to the AMB treatment varied from 15 to 90%, and increased with temperature, but decreased with N status. Nevertheless, the absolute photosynthetic response to EL increased with shoot N status. The relative increase in the instantaneous response of A(max) to elevated [CO2] was about 20% higher than the long-term response, i.e., there was downward acclimation in Amax in response to elevated [CO2]. The photosynthetic temperature optimum increased 4 degrees C with either a short- or a long-term increase in [CO2]. The bag treatment itself increased A(max) by approximately 16% and the temperature optimum of A(max) by approximately 3 degrees C. PMID- 11498341 TI - Growth and dry-matter partitioning of young Populus trichocarpa in response to carbon dioxide concentration and mineral nutrient availability. AB - Young individuals of a single black cottonwood (Populus trichocarpa Torr. & Gray) clone were raised for three growing seasons in whole-tree chambers and exposed to either ambient or elevated atmospheric carbon dioxide concentration ([CO2]), with either a high or a low mineral nutrient supply, in a factorial experimental design. Nutrient availability had a larger effect on growth and dry matter partitioning than did [CO2]. Total biomass did not differ significantly with CO2 treatment when nutrient availability was low. However, elevated [CO2] increased whole-plant biomass by 47% in the high nutrient availability treatment. Carbon dioxide enrichment reduced leaf area ratio and specific leaf area significantly, but had no significant effect on mean leaf size or leaf mass ratio. Root mass ratio was significantly increased by elevated [CO2] at low, but not at high nutrient availability. A modified "demographic harvesting approach" made possible the retrospective estimation of stem and branch dry masses for different years. The relative growth rates of stem and branch were significantly enhanced by elevated [CO2] with high, but not with low nutrient availability. Canopy productivity index (CPI), i.e., the amount of stem and branch wood produced annually per unit leaf area, was raised 12% by elevated [CO2] when nutrient availability was high, but was reduced when nutrient availability was low, because of increased below ground allocation. PMID- 11498342 TI - Leaf morphology and photosynthetic adjustments among deciduous broad-leaved trees within the vertical canopy profile. AB - Photosynthetic acclimation of deciduous broad-leaved tree species was studied along a vertical gradient within the canopy of a multi-species deciduous forest in northern Japan. We investigated variations in (1) local light regime and CO2 concentration ([CO2]), and (2) morphological (area, thickness and area per mass), biochemical (nitrogen and chlorophyll concentrations) and physiological (light saturated photosynthetic rate) attributes of leaves of seven major species on three occasions (June, August and October). We studied early successional species, alder (Alnus hirsuta (Spach) Rupr.) and birch (Betula platyphylla var. japonica (Miq.) Hara); gap phase species, walnut (Juglans ailanthifolia Carriere) and ash (Fraxinus mandshurica var. japonica Rupr.); mid-successional species, basswood (Tilia japonica (Miq.) Simonk.) and elm (Ulmus davidiana var. japonica (Rehd.) Nakai); and the late-successional species, maple (Acer mono Bunge). All but maple initiated leaf unfolding from the lower part of the crown. The [CO2] within the vertical profile ranged from 320-350 ppm in the upper canopy to 405 560 ppm near the ground. The lowest and highest ambient [CO2] occurred during the day and during the night, respectively. This trend was observed consistently during the summer, but not when trees were leafless. Chlorophyll concentration was positively related to maximum photosynthetic rate within, but not among, species. Leaf senescence started from the inner part of the crown in alder and birch, but started either in the outer or top portion of the canopy of ash, basswood and maple. Chlorophyll (Chl) to nitrogen ratio in leaves increased with decreasing photon flux density. However, Chl b concentration in all species remained stable until the beginning of leaf senescence. Maximum photosynthetic rates observed in sun leaves of early successional species, gap phase or mid successional species, and late successional species were 12.5-14.8 micromol m(-2) s(-1), 4.1-7.8 micromol m(-2) s(-1) and 3.1 micromol m(-2) s(-1), respectively. PMID- 11498343 TI - Foliar morphological and physiological plasticity in Picea abies and Abies alba saplings along a natural light gradient. AB - The role of morphological versus physiological foliar plasticity in the capacity for, and mechanisms of, photosynthetic acclimation was assessed in Picea abies (L.) Karst. and Abies alba Mill. saplings in a forest gap-understory light gradient (relative irradiance, RI, ranging from 0.02 to 0.32). The species investigated showed a similar foliar morphological plasticity along the light gradient, at both the needle level (through alteration in leaf dry mass per area) and the shoot level (through alteration in the silhouette area ratio, e.g., shoot silhouette to projected needle area ratio). In both species chlorophyll (Chl) concentration on a mass basis decreased at increasing RI, but was independent of RI when expressed on an area basis. In contrast, leaf N concentration on a mass basis was independent of RI, but was positively influenced by RI when expressed on an area basis. The parameters describing photosynthetic performance at low light (dark respiration rate, apparent quantum yield and light compensation point) suggest that Abies alba was better suited to maintain a positive carbon balance in shaded conditions. By contrast, parameters describing biochemical capacity at high light (maximum electron transport rate, Jmax and maximum ribulose-1,5-biphosphate carboxylation capacity, Vcmax) indicate that only Picea abies was capable of acclimating physiologically to high photosynthetic photon flux densities (PPFDs) by increasing nitrogen partitioning to Rubisco and Vcmax/mass by increasing RI. These results support the hypothesis that interspecific differences in nitrogen partitioning within the photosynthetic apparatus may provide a mechanistic basis for species separation along a light gradient. The differences in photosynthetic plasticity observed are likely to influence regeneration patterns and habitat breadth of the species investigated. The limited ability of Abies alba saplings to exploit high-light conditions may be a competitive disadvantage in large canopy gaps and thus limit recruitment of this species to small gaps. PMID- 11498344 TI - Carbon dioxide exchange in Norway spruce at the shoot, tree and ecosystem scale. AB - Net CO2 exchange in a 35-year-old boreal Norway spruce (Picea abies (L.) Karst.) forest in northern Sweden was measured at the shoot (NSE), tree (NTE) and ecosystem levels (NEE) by means of shoot cuvettes, whole-tree chambers and the eddy covariance technique, respectively. We compared the dynamics of gross primary production (GPP) at the three levels during the course of a single week. The diurnal dynamics of GPP at each level were estimated by subtracting half hourly or hourly model-estimated values of total respiration (excluding light dependent respiration) from net CO(2) exchange. The relationship between temperature and total respiration at each level was derived from nighttime measurements of NSE, NTE and NEE over the course of 1 month. There was a strong linear relationship (r2 = 0.93) between the hourly estimates of GPP at the shoot and tree levels, but the correlation between shoot- and ecosystem-level GPP was weaker (r2 = 0.69). However, the correlation between shoot- and ecosystem-level GPP was improved (r2 = 0.88) if eddy covariance measurements were restricted to periods when friction velocity was > or = 0.5 m s(-1). Daily means were less dependent on friction velocity, giving an r2 value of 0.94 between shoot- and ecosystem-level GPP. The correlation between shoot and tree levels also increased when daily means were compared (r2 = 0.98). Most of the measured variation in carbon exchange rate among the shoot, tree and ecosystem levels was the result of periodic low coupling between vegetation and the atmosphere at the ecosystem level. The results validate the use of measurements at the shoot and tree level for analyzing the contribution of different compartments to net ecosystem CO2 exchange. PMID- 11498345 TI - Daily and seasonal patterns of carbon and water fluxes above a north Australian savanna. AB - Daily and seasonal fluxes of carbon dioxide and water vapor above a north Australian savanna were recorded over a complete dry season-wet season annual cycle using the eddy covariance technique. Wet season rates of photosynthesis and transpiration were larger than those measured in the dry season and were dominated by the presence of the grassy understory. As the dry season progressed and the grass understory died, ecosystem rates of assimilation and water vapor flux declined substantially. By the end of the dry season, canopy assimilation and evapotranspiration rates were 20-25% of wet season values. Assimilation was light saturated in the dry season but not in the wet season. Stomatal control of transpiration increased between the wet and dry season. This was revealed by the decline in the slope of E with increasing leaf-to-air vapor pressure difference (D) between wet and dry seasons, and also by the significant decrease in the ratio of boundary to canopy conductance observed between the wet and dry seasons. A simple pan-tropical modeling of leaf area index or wet season canopy CO2 flux was undertaken. It was shown that with readily available data for foliar N content and the ratio of rainfall to potential evaporation, leaf index and wet season canopy CO2 flux can be successfully estimated for a number of tropical ecosystems, including north Australian savannas. PMID- 11498346 TI - Crown structure and leaf area index development in thinned and unthinned Eucalyptus nitens plantations. AB - The crown structure of Eucalyptus nitens (Deane & Maiden) Maiden 6 years after thinning, and the development of stand leaf area index both immediately and 6 years after thinning, were investigated. Thinning did not alter branch angle, branching density or the relationship between branch size and branch leaf area. However, larger branches were found in the lower crown of thinned trees and the increase in leaf area as a result of thinning occurred on the northern aspect of the crown. The vertical distribution of leaf area in unthinned trees was skewed toward the top of the crown and correlated with live crown ratio. The vertical distribution of leaf area in thinned trees tended to be less skewed and was unrelated to tree size or dominance. Leaf area index, as estimated from light interception measurements, increased at a constant rate soon after thinning regardless of residual stocking. In the longer term, residual stocking had a strong influence on leaf area increase per tree and was correlated with changes in crown length. PMID- 11498347 TI - Effects of thinning on wood production, leaf area index, transpiration and canopy interception of a plantation subject to drought. AB - We conducted thinning trials in a 5-year-old Eucalyptus globulus ssp. globulus Labill plantation near Warrenbayne, northeastern Victoria, Australia, where soil salinization and waterlogging are common, and assessed treatment effects on tree growth, water use and survival. Half-hectare plots were thinned from the original density of 1100 stems ha(-1) to densities of 800, 600 and 400 stems ha(-1), and stem diameter increment, leaf area index, transpiration, canopy interception and depth of tree water source monitored for 21 months. Two drought periods occurred during the study, rainfall was 30% below the long-term average and there was severe mortality in all three plots. Analysis of deuterium abundance in soil and xylem water indicated that the trees accessed water only from the top meter of the soil profile. Transpiration rates were higher in the most heavily thinned plot than in the least thinned plot, which underwent a reduction in basal area during the study. The most heavily thinned plot increased in basal area by 10% during the study. Edge trees had significantly greater diameters than trees from the middle of the plots. PMID- 11498348 TI - Interception loss, throughfall and stemflow chemistry in pine and oak forests in northeastern Mexico. AB - Interception loss, gross precipitation, throughfall and stemflow solution chemistry beneath pine (Pinus pseudostrobus Lindl.), oak (Quercus sp.) and pine oak natural forest canopies in northeastern Mexico were measured. Coefficients of variation for throughfall were 12% in pine and oak canopies and 17% in the mixed pine-oak canopy. The variability of stemflow averaged 66, 126 and 73% for pine, oak and the mixed pine-oak canopies, respectively. Linear regression analysis of net versus gross precipitation for the three canopies showed highly significant correlations (r = 0.974-0.984). Total precipitation during the experimental period was 974 mm and estimated interception loss was 19.2, 13.6 and 23% for the pine, oak and pine-oak canopies, respectively. Stemflow did not occur following rainfall events of less than 4 mm and, in all canopies, stemflow represented a minimal proportion of gross precipitation (0.60, 0.50 and 0.03% for pine, oak and pine-oak, respectively). Throughfall pH in pine (6.2), oak (6.3) and pine-oak (6.3) canopies was significantly more acidic than gross precipitation (6.6). Stemflow pH ranged from 3.7 (pine) to 6.0 (oak). The pine-oak canopy registered the highest throughfall and stemflow electrical conductivities, 104 and 188 microS cm(-1), respectively. Net nutrient leaching of K, Mg, Na, Fe, Mn and Zn was significantly higher from the pine-oak canopy than from the pure pine and oak canopies. Mean depositions of Ca and Cu in throughfall behaved similarly among the three types of canopies. A greater proportion of Zn in gross precipitation was absorbed by the oak canopy than by the pine and pine-oak canopies. Enrichment factors beneath the pine-oak canopy relative to gross precipitation varied from 1.2 to 3.2 for macro-nutrients (Ca, K, Mg and Na) and from 1.4 to 3.1 for micro nutrients (Cu, Fe, Mn and Zn). Stemflow depositions of Ca, K, Mg and Cu were higher in the pine-oak canopy, whereas stemflow depositions of Na, Fe, Mn and Zn were higher in the pine canopy. PMID- 11498349 TI - 10th Conference on DNA Topoisomerases in therapy. AB - The 10th Conference on DNA Topoisomerases in Therapy 6-8 October 1999 in Amsterdam, The Netherlands) covered basic research on DNA topoisomerases and aspects of topoisomerase-directed therapy. The understanding of basic aspects of enzyme functions and structures was discussed throughout the meeting, as this knowledge is fundamental to further developments of new and more effective therapeutic approaches. Several new crystal structures were presented, and implications for function and interaction with DNA and drugs were discussed. Knock-out mice for various topoisomerase genes have been produced and genes have been shown to differ in importance for development and survival. The interaction of topoisomerases with other proteins involved in DNA metabolism, chromosome stability and physiology were discussed. The main focus for cancer therapy was on camptothecins (CPT) and related compounds stabilizing covalent DNA-intermediates of topoisomerase I. Reports on recent clinical trials of first-generation, water soluble CPT-analogs (topotecan and irinotecan) confirmed earlier findings of activity in several solid tumors and hematological malignancies. Improvements in efficacy and toxicity profiles are being sought in orally absorbable compounds and other drug formulations (e.g. in liposomes). Several new CPT-analogs at preclinical stages of development might also provide a greater stability of the lactone ring, higher DNA-binding affinity, and improved water solubility. New drugs have also been developed from a number of new non-CPT compounds, which inhibit the activity of DNA-topoisomerases but do not stabilize the DNA-linked form of the enzymes. Another focus of the meeting was on therapeutic targeting of microbial DNA topoisomerases. The antibiotic potential of the quinolones has been extended to gram-positive pathogens, particularly Streptococcus pneumoniae. The cloning and biochemical characterization of the DNA-topoisomerases of eukaryotic parasites such as Plasmodium falciparum or Candida albicans have been completed and the search for specific inhibitors targeting these enzymes is under way. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498350 TI - Molecular mechanisms of resistance to taxanes and therapeutic implications. AB - The mechanism of resistance to taxanes has not been fully elucidated. Since Taxol is a substrate for P-glycoprotein, overexpression of this transport system is recognized as a relevant mechanism of resistance. Additional mechanisms include changes of microtubule structure and/or composition resulting in reduced drug binding to the target. Current efforts are directed at clarifying the role of cellular response to drug-induced damage to cytoskeleton and mitotic spindle. Downstream events, such as control of cell cycle progression and regulation of cell death pathways, are likely to play a relevant role in cellular sensitivity to antimicrotubule agents. The identification of resistance factors and critical determinants of antitumor efficacy of microtubule-stabilizing agents is essential to (i) improve their therapeutic efficacy; and (ii) to design non-cross-resistant compounds. The present review discusses the possible therapeutic implications of the recent progress in the field of resistance to taxanes. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498351 TI - Mining bacterial cell wall biosynthesis with new tools: multitarget screens. AB - The cytoplasmic steps of peptidoglycan synthesis remain underexplored for the discovery of novel antibiotics. Pathway screens are well suited to screen for novel inhibitors because several targets are tested at once. Whole-cell based assays are easier to set up but they tend to favor identification of compounds that inhibit enzymes that are at the limiting rate in the pathway. Screens that use purified enzymes are optimized to detect with similar probability inhibitors of any of the targets in the pathway. This approach is being used to identify novel promising molecules. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498352 TI - Active transport of siderophore-mimicking antibacterials across the outer membrane. AB - The outer membrane of gram-negative bacteria forms a permeability barrier that usually reduces antibiotic access to intracellular targets and renders gram negative bacteria less susceptible to antibiotics than gram-positive bacteria, which lack an outer membrane. However, gram-negative bacteria become highly susceptible to antibiotics that are actively transported across the outer membrane. Some antibiotics use active transport systems of substrates with which they share structural features. Examples are naturally occurring sideromycins and synthetic derivatives of Fe(3+)-siderophores, which are taken up across the outer membrane by transport systems for Fe(3+)-siderophores. A well-studied example is albomycin, which has structural similarities to the natural substrate ferrichrome; albomycin and ferrichrome are both transported by the FhuA protein. A semisynthetic rifamycin derivative, CGP 4832, is also taken up by the FhuA transport protein, although its structure is completely different from that of ferrichrome. The crystal structures of FhuA with bound ferrichrome, albomycin, or rifamycin CGP 4832 reveal that the three compounds occupy the same site on FhuA; this site is accessible from the growth medium by a surface cavity that accommodates the antibiotic moieties. There is a rather strict stereochemical requirement for the portion that fits into the active site of FhuA, but a rather large tolerance regarding the portion that is located in the cavity. These data provide precise structural information for the design of highly active antibiotics composed of an antibiotically active moiety connected by a linker to a transported carrier. A number of Fe(3+)-siderophore carriers of the hydroxamate and catechol type linked to antibiotics have been isolated from microbes and synthesized; their superior efficacy has been demonstrated in vitro and in mice. Although none have been therapeutically employed, it is proposed that this alternative method of synthesizing useful antibiotics should be tested in light of the increasing problem of resistant pathogens. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498353 TI - Photoaffinity analogs for multidrug resistance-related transporters and their use in identifying chemosensitizers. AB - A major obstacle in cancer treatment is the development of resistance to multiple chemotherapeutic agents in tumor cells. The hallmark of this multidrug resistance (MDR) is overexpression of the MDR 1 P-glycoprotein or the multidrug resistance protein MRP1. It is well documented that these proteins confer MDR in cancer cells. Much evidence indicates that control of intracellular drug levels in MDR cells is determined by P-glycoprotein or MRP, and therefore these proteins are suitable targets for identifying MDR-reversing agents (MDR modulators). We originally explored the drug-binding ability of P-glycoprotein by synthesizing and using radioactive photoaffinity analogs of vinblastine. Since our initial discovery that P-glycoprotein binds to vinblastine photoaffinity analogs, many P glycoprotein- and MRP-specific photoaffinity analogs have been developed. In this review, photoaffinity analogs which specifically bind to P-glycoprotein or MRP are discussed. Moreover, utilizing these photoprobes to identify, characterize and localize the drug binding sites of P-glycoprotein and MRP is described. Using P-glycoprotein-specific photoaffinity analogs in combination with site-directed antibodies to several domains of this protein has allowed the localization of the general binding domains of some of the cytotoxic agents an MDR modulators on P glycoprotein. However, the molecular architecture of the drug binding sites, their exact location on the P-glycoprotein molecule, and the total number of the drug binding sites remain to be determined. This review discusses recent advances in delineating the structure of the drug-binding sites of P-glycoprotein. Moreover, novel MRP1 photoaffinity analogs are reviewed. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498354 TI - Tumor cell resistance to DNA topoisomerase II inhibitors: new developments. AB - DNA topoisomerases are critical enzymes involved in replication, transcription, chromatin assembly and other aspects of DNA metabolism. They are also the targets of important anticancer drugs. The type II topoisomerases are specific targets of drug classes that comprise complex-stabilizing (epipodophyllotoxins, anthracyclines) and catalytic (merbarone, bisdioxopiperazines) inhibitors. In this review, we update our current knowledge of resistance to the antitumor inhibitors of the type II DNA topoisomerases, with special emphasis on the catalytic inhibitors, since novel catalytic inhibitor resistant cell lines have only recently been described. Resistance to topoisomerase II inhibitors can manifest as decreased or increased expression of or mutation in the topoisomerase II genes. However, the tumor cell's response to exposure to these inhibitors involves more than the target enzyme, and these other responses are a major focus of this review. Such cellular changes are associated with and may contribute to the drug resistance phenotype. They involve decreased drug accumulation due to expression of membrane 'pump' proteins, altered cytotoxic signaling through stress-activated protein kinases, and alterations in apoptosis and cell cycle proteins (e.g. Bcl-2, Bax, p53, Rb). While it is evident that mutation in or altered expression of the topoisomerase II genes are sufficient to confer resistance to topoisomerase inhibitors, it is not clear whether the other changes are a consequence of the selection or a response to the cytotoxic insult, nor is it clear how these other cellular changes contribute to the drug resistance phenotype. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498355 TI - Molecular aspects of azole antifungal action and resistance. AB - During the past three decades azole compounds have been developed as medical and agricultural agents to combat fungal diseases. During the 1980s they were introduced as orally active compounds in medicine and the number of such azole drugs is likely to expand in the near future. They represent a successful strategy for antifungal development, but as the incidence of fungal infection has increased coupled to prolonged use of the drugs, the (almost) inevitable emergence of resistance has occurred. This was after resistance had already been encountered as a serious problem in the field, where a larger number of azole fungicides had been employed commercially. In this review the molecular basis of how azoles work is discussed together with how fungi overcome the inhibitory effect of these compounds: through alterations in the primary target molecule (cytochrome P45051; Erg11p; sterol 14alpha-demethylase); through drug efflux mechanisms and through a suppressor mechanism allowing growth on 14-methylated sterols. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498356 TI - Regulation of pleiotropic drug resistance in yeast. AB - This review focuses on the molecular mechanisms involved in the regulation of multiple drug resistance in the model yeast Saccharomyces cerevisiae and the pathogenic fungus Candida albicans. Recent developments in the study of the transcription factors Pdr1p, Pdr3p and Yap1p are reported. Understanding the molecular basis leading to multiple drug resistance is a prerequisite for the development of new antifungal therapeutics. Copyright 1999 Harcourt Publishers Ltd. PMID- 11498357 TI - Regular Article. PMID- 11498358 TI - Editorial. PMID- 11498359 TI - Telomerase as a potential anticancer target: growth inhibition and genomic instability. AB - Stabilization of telomere length in chromosomes by an RNA-dependent DNA polymerase (telomerase) appears to be responsible for the replicative immortality of cancer cells. These findings provide the rational basis for generating experimental models to develop anti-telomerase drugs. However, there is conflicting evidence in the literature about the outcome of telomerase inhibition. While tumor cytostatic and cytotoxic effects associated with telomerase inhibition have been described, absence of telomerase has been associated with genetic instability and tumor development. Therefore, a therapeutic strategy based on telomerase inhibition will likely have to cope with problems related to innate or acquired mechanisms of drug resistance and possibly to therapy-related tumors. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498360 TI - Hypoxic cytotoxic agents: a new approach to cancer chemotherapy. AB - Many of the cells in solid tumors are hypoxic. This makes them resistant to killing by both radiotherapy and most commonly used anticancer drugs. The resistance towards anticancer drugs is because hypoxia induces a non proliferative state as well as the fact that the hypoxic cells are furthest from the blood vessels in the tumor (and hence the source of drug). However, a new class of drugs - hypoxic cytotoxins can, by killing these hypoxic cells, not only overcome this resistance, but also exploit hypoxia thereby converting it into a therapeutic advantage. The first drug to be tested in the clinic purely for its activity towards hypoxic cells, tirapazamine, is described. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498361 TI - Antifungal susceptibility testing of yeasts: uses and limitations. AB - With recent developments in the field of mycology, such as increased incidence of fungal infections, the introduction of newer, safer antifungals, and the emergence of resistance, the need for clinically relevant antifungal susceptibility testing methods is obvious. Studies performed over the past decade have allowed the National Committee for Clinical Laboratory Standards Subcommittee on Antifungal Testing to achieve consensus on a new standardized broth dilution method for in vitro susceptibility testing of yeasts (NCCLS M27 A). Once the reproducibility of the M27-A document was established, tentative breakpoints for fluconazole and itraconazole were derived. The availability of a standardized procedure for determining the minimum inhibitory concentrations (MICs) of antifungal agents is an important tool in drug discovery and development. In addition, it provides means for detection of resistant strains and, in the case of oropharyngeal candidiasis, means for patient management. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498362 TI - Apoptosis induction by arsenic: mechanisms of action and possible clinical applications for treating therapy-resistant cancers. AB - Arsenic, a known carcinogen, may be useful in cancer treatment. Arsenic may be effective in counteracting drug resistance because it appears to induce apoptosis in tumor cells independently of p53 activation, thereby allowing it to be directed against p53-defective cancers. The role of MAP kinases in arsenic induced apoptosis in tumor cells is important and may be influenced by reactive oxygen species or glutathione. This review focuses on recent findings from this and other laboratories regarding the mechanism(s) of arsenic-induced apoptosis in tumor cells and considers their relevance in the clinical treatment of therapy resistant cancers. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498363 TI - Biochemical mechanisms involved in overcoming HIV resistance to nucleoside inhibitors of reverse transcriptase. AB - The development of drug combinations that act effectively against both wild-type and mutated resistant forms of HIV-1 reverse transcriptase (RT) is a major goal in management of HIV disease. Recent studies have shown that resistance to different nucleoside analog RT inhibitors (NRTIs), an important class of anti viral drugs, can result in different amino acid substitutions in close proximity to the dNTP binding pocket of the enzyme. Some of these mutations have been shown to cause cross- or multiple resistance among various members of this family of inhibitors. In contrast, certain combinations of amino acid substitutions can sometimes lead to increased drug susceptibility and may also result in resensitization of formerly resistant viruses. A biochemical understanding of these complex viral phenotypes may be of major importance in regard to development of novel chemotherapeutic strategies that can act at the level of drug-resistant mutated enzymes. In this review, we discuss several principles that help to explain the increased susceptibility and resensitization to some antiviral agents used in the context of combination treatment. The conclusions are largely based on our current understanding of mechanisms involved in drug resistance to 3TC and AZT. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498364 TI - pH and drug resistance in tumors. AB - Uptake of weakly ionizing drugs by tumours is greatly influenced by the interstitial and intracellular pH, and the ionization properties of the drug. Extracellular pH in tumors is acidic, while the intracellular pH is in the neutral-to-alkaline range. Tumors of the bladder, kidney and gastrointestinal system in particular are exposed to extremes of pH. Strategies for exhancing and exploiting acid-outside plasmalemmal pH gradients to drive the uptake of weak acid drugs into tumors are discussed, as are techniques for alkalinizing tissues to improve response to weak base drugs. The participation of acidic intracellular vesicles in non-specific drug resistance is explored. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498365 TI - The pH difference between tumor and normal tissue offers a tumor specific target for the treatment of cancer. PMID- 11498366 TI - Workshop on ABC Transporters and Human Diseases. PMID- 11498367 TI - 3rd International Conference on Therapies for Viral Hepatitis. AB - This meeting was attended by several hundred delegates and covered, in lecture and poster presentations, recent developments in basic, applied and clinical research as they concern antiviral drug development for hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Below are summaries of presentations that are relevant to the development of antiviral drug resistance. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498368 TI - Molecular targets and cancer therapeutics: discovery, development and clinical validation. AB - The AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics held in Washington, DC on 16-19 November 1999 provided a forum for cancer research clearly showing evolution of a target and mechanism-driven science. The notion of the tumor as a tissue composed of heterogeneous populations of normal and abnormal cells as viable targets is coming to the fore with the advent of agents directed toward non-malignant cell targets. Growth control rather than eradication as a treatment strategy for malignant disease is being tested preclinically and clinically. Among targets, kinases are in the lead with nuclear, cytoplasmic and membrane kinases being selectively inhibited by small molecules and macromolecules. First generation tumor vasculature-directed agents are progressing through early clinical studies. The interest in tumor vasculature as a target has renewed interest in imaging technology to discern biological effect and in tumor hypoxia. This has resulted in elucidation of molecular responses triggered by a low oxygen environment. Challenges remain in the areas of cellular and immune therapies. Dendritic cell-based vaccines are being tested preclinically in many systems. Interleukin-12 is proceeding through clinical trials. Apoptosis-protective molecules such as bcl-2, and apoptosis stimulating molecules such as bax, are being pursued as targets for inhibition and activation, respectively. Finally, methods and technology to aid in the identification of new targets were highlighted. This perspective, while it is by no means an exhaustive review of the presentations, brings forward some of the main topics and interests that are current in cancer research. Targets were the topic but methods of target identification and the need for increased chemical diversity to selectively focus agents to targets with small differences were also major topics of discussion. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498369 TI - Molecular Approaches to Malaria 2000. AB - For more than 20 years now, Australia has been officially free of endemic malaria, but this devastating disease once again made a major impact on the continent in February 2000 when Melbourne hosted Australia's first major international conference on 'Molecular Approaches to Malaria' (Lorne, Australia, 2-5 February 2000). The global research effort toward our increased understanding of the pathogenesis and control of malaria in the post-genomics era was discussed and debated at length over 4 days packed with science encompassing molecular biology, cell biology, clinical studies, genomics, vaccines and pathogenic mechanisms. More than 260 researchers from 18 countries worldwide participated in this interdisciplinary meeting which comprised 57 oral presentations and 122 posters. Here we summarize some presentations pertinent to the field of drug action and resistance. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498370 TI - Turning p53 on or off: either way may treat cancer. AB - The p53 tumor suppressor gene is likely the most commonly mutated tumor suppressor gene in human cancer. Its functions include modulation of both cell cycle arrest and apoptosis. Animal models and human clinical data suggest that in some settings, p53 may be prognostically significant, reflecting its role as a key regulator of cell death during cancer therapy. Two recent strategies have been proposed to exploit p53's unique death-regulating activity in opposite directions and improve cancer treatment. One approach seeks to inhibit p53 in normal cells thereby diminishing therapy-related, p53-dependent toxicity. The other utilizes a peptide derived from the C-terminus of p53 to activate wild-type or mutant p53 proteins, triggering apoptosis with selectivity for transformed cells. These novel approaches hold promise for targeting p53 in cancer therapy and may shed light on mechanisms underlying the role of p53 in cancer cell survival. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498371 TI - Definitive proof for a role of pfmdr 1 in quinoline resistance in Plasmodium falciparum. PMID- 11498372 TI - Cyclin-dependent kinases inhibitors as potential anticancer, antineurodegenerative, antiviral and antiparasitic agents. AB - Cyclin-dependent kinases (CDKs) play a key role in the cell division cycle, in neuronal functions, in transcription and in apoptosis. Intensive screening with these kinases as targets has lead to the identification of highly selective and potent small - molecule inhibitors. Co-crystallization with CDK2 shows that these flat heterocyclic hydrophobic compounds bind through two or three hydrogen bonds with the side chains of two amino acids located in the ATP-binding pocket of the kinase. These inhibitors are anti-proliferative; they arrest cells in G1 and in G2/M phase. Furthermore they facilitate or even trigger apoptosis in proliferating cells while they protect neuronal cells and thymocytes from apoptosis. The potential use of these inhibitors is being extensively evaluated for cancer chemotherapy and also in other therapeutic areas: neurology (Alzheimer's disease), cardiovascular (restenosis, angiogenesis), nephrology (glomerulonephritis), parasitology (Plasmodium, Trypanosoma, Toxoplasma, etc.) and virology (cytomegalovirus, HIV, herpes virus). Copyright 2000 Harcourt Publishers Ltd. PMID- 11498373 TI - Treatment strategies for Aspergillus infections. AB - Infections caused by Aspergillus species consist of many different disease presentations, ranging from relatively benign asthma in atopic disease to life threatening systemic invasive infections. The spectrum of disease manifestations is determined by a combination of genetic predisposition, host immune system defects, and virulence of the Aspergillus species. For the purposes of this discussion, we will address three principal entities: invasive aspergillosis, both primary and disseminated, pulmonary aspergilloma, and allergic bronchopulmonary aspergillosis. Amphotericin B is the standard of treatment for severe Aspergillus infections, despite the fact that mortality in these patients remains high. Alternative therapies such as combination regimens and itraconazole also have efficacy against Aspergillus infections. We discuss the role of current therapies, the potential role of drugs in development, and the results of ongoing research with combination and immunotherapies. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498374 TI - Regulation of MDR1 gene expression: emerging concepts. AB - Drug resistance genes, such as MDR1, involved in drug efflux, and their regulation have been the subject of intense research efforts in the past 10 years. Many factors and cellular signalling pathways play a role in the regulation of MDR1 gene expression. Commonly used chemotherapeutic agents activate in vitro and in vivo general stress response pathways, potential targets of which include MDR1 and other drug resistance genes. The contribution of these agents to the emergence of drug-resistant tumour cells is of concern. Recent evidence points to a role for the epigenetic regulation of MDR1 gene expression. The identification of key components in the DNA methylation/chromatin system of gene regulation may in time lead to more informed and targeted approaches to treating drug-resistant tumours. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498375 TI - b-Lactamase inhibitors. AB - The use of beta-lactamase inhibitors in combination with a beta-lactamase susceptible antibiotic is a useful strategy to rescue otherwise good antibiotics from failure. However, recent years have seen a rise in the numbers of beta lactamases that are insensitive to the available beta-lactamase inhibitors. This review summarizes of the mechanisms of action of the principal types of inhibitors and the ways in which beta-lactamase are thought to develop resistance towards them. Ten general classes of inhibitors are reviewed, especially those of therapeutic importance (clavulanic acid, penam sulfones and carbapenems). Copyright 2000 Harcourt Publishers Ltd. PMID- 11498377 TI - A new resistance mechanism to cisplatin? PMID- 11498376 TI - Cancer research 2000: drug resistance, new targets and drugs in development. AB - The 91st Annual Meeting of the American Association for Cancer Research (AACR) provided the latest developments in cancer research. We highlight here presentations on resistance mechanisms (tumor microenvironment, efflux pumps, apoptosis), new targets and drugs in development (signal transduction, cell cycle, apoptosis, microtubules, topoisomerases) and new technologies (the cancer genome anatomy project, the human genome project, proteomics). The importance of signaling pathways or networks ('cell context') in defining the role of particular proteins was brought up in several presentations. A summary of specific drugs in development is also included in this report. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498378 TI - The role of reductive enzymes in cancer cell resistance to mitomycin C. AB - Mitomycin C (MMC) is bioreductively activated to DNA binding species via complex chemical pathways involving a common hydroquinone intermediate. A recent publication by Belcourt et al. (1999) has revealed that the bacterial mitomycin C resistance protein (MCRA) acts as a unique hydroquinone oxidase converting this reactive intermediate back to the parent drug in the presence of molecular oxygen, preventing the formation of cytotoxic interstrand DNA crosslinks. It was argued that a mechanism analogous to MCRA may be responsible for the often observed phenomenon of aerobic drug resistance that develops in vitro to MMC in human cancer cell lines. Altered expression of activating reductase enzymes, which usually accompanies aerobic drug resistance, was claimed to be of lesser importance. Therefore, the role of reductases in MMC drug resistance has been reviewed. While it is clear from numerous studies that lowered reductase expression can in certain situations produce drug resistance, simple correlations between a specific enzyme and chemosensitivity generally do not hold due to the complex functional and regulatory interplay that exists among the different activating enzymes and detoxification systems. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498379 TI - In vivo veritas: Bcl-2 and Bcl-X(L)mediate tumor cell resistance to chemotherapy. AB - Apoptosis is cellular suicide, the functional opposite of mitosis. It may play an important role in tissue growth control and removal of damaged and premalignant cells. The fact that diverse chemotherapeutic agents induce apoptosis, while they engage different intracellular targets and cause DNA damage, raises a concern that tumors resistant to chemotherapy are unable to initiate the apoptotic process. The anti-apoptotic Bcl-2 family proteins, Bcl-2 and Bcl-X(L), play an important role in the regulation of apoptotic cell death. Bcl-2 and Bcl-X(L)have been reported to confer chemotherapy resistance in short-term survival assays in vitro. However, they failed to provide a long-term clonogenic survival advantage. Thus, the role of anti-apoptotic Bcl-2 and Bcl-X(L)on chemotherapy resistance in vivo remains unclear. In vivo, tumor cells receive survival signals from the extracellular microenvironment. Since the microenvironmental factors have been reported to modulate the expression and function of Bcl-2 family proteins, Bcl-2 and Bcl-X(L)might be associated with the chemotherapy resistance in vivo through the influence of these factors. Consistent with this hypothesis, several investigators have recently reported that the sensitivity to chemotherapy in in vitro clonogenic assays did not correlate with that in in vivo tumor models. The lack of microenvironmental factors might cause the discrepancy between in vitro clonogenic growth and in vivo tumor growth. These results suggest that Bcl-2 and Bcl-X(L)could contribute to chemotherapy resistance in vivo, along with already defined drug resistance mechanisms (i.e. P-glycoprotein, MRP). Therapies aimed at suppressing the expression and function of Bcl-2 and Bcl-X(L)or at intercepting microenvironmental factors might successfully overcome chemotherapy resistance. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498380 TI - Sulfonamide resistance: mechanisms and trends. AB - Sulfonamides were the first drugs acting selectively on bacteria which could be used systemically. Today they are infrequently used, in part due to widespread resistance. The target of sulfonamides, and the basis for their selectivity, is the enzyme dihydropteroate synthase (DHPS) in the folic acid pathway. Mammalian cells are not dependent on endogenous synthesis of folic acid and generally lack DHPS. Instead, they have a folate uptake system which most prokaryotes lack. Laboratory mutants in the dhps (folP) gene can be easily isolated and show a trade off between sulfonamide resistance and DHPS enzyme performance. Clinical resistant mutants, however, have additional compensatory mutations in DHPS that allow it to function normally. In many pathogenic bacteria sulfonamide resistance is mediated by the horizontal transfer of foreign folP or parts of it. Clinical resistance in gram-negative enteric bacteria is plasmid-borne and is effected by genes encoding alternative drug-resistance variants of the DHPS enzymes. Two such genes, sul1 and sul2, have been sequenced and are found at roughly the same frequency among clinical isolates. Remarkably, the corresponding DHPS enzymes show pronounced insensitivity to sulfonamides but normal binding to the p aminobenzoic acid substrate, despite the close structural similarity between substrate and inhibitor. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498381 TI - Clinical applications of telomerase in cancer treatment. AB - Telomerase activity has been found in most cancer cells, but not in the majority of normal differentiated tissues. Therefore, telomerase has been considered a relatively selective and widely expressed tumor marker to be used as a diagnostic tool, and in some cases, as a potential prognostic indicator. Telomerase activity can also be used to evaluate chemosensitivity of neoplastic cells obtained from cancer patients, by measuring residual telomerase activity after drug treatment. Finally, telomerase has been considered to represent a suitable target for designing new anticancer strategies. This review focuses on present and future clinical applications of telomerase studies in cancer management. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498382 TI - Liposomal formulations of anticancer drugs: selectivity and effectiveness. AB - After a delay of roughly 30 years, liposomes are becoming important as drug targeting systems especially in anticancer therapy. Two liposomal anthracycline formulations are now available for cancer treatment and more are in preclinical tests or in clinical trials. This review summarizes the possibilities to improve the anticancer activity of certain drugs by entrapment in liposomes. We will briefly examine the problems and limitations of liposomal systems, from a pharmacokinetic as well as from a manufacturing viewpoint. Moreover, the anticancer properties of the two new liposomal anthracycline formulations will be discussed with regard to anticancer activity and side effects and compared with the properties of the respective free drugs. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498383 TI - beta -Lactamases: which ones are clinically important? AB - The introduction of a large array of beta-lactam antibiotics has spawned the emergence of an even larger variety of beta-lactamases designed to confer resistance to these agents. beta-lactamases are produced by both gram-positive and gram-negative bacteria, but their clinical importance is far greater among the gram-negatives. The virtual explosion in our knowledge about the variety of these enzymes can often create confusion and frustration among those not well versed in the field. In this paper, we attempt to focus the discussion of beta lactamases on those enzymes that are of the greatest clinical importance, the Ambler Class A and C enzymes. We also discuss the growing importance of the Ambler Class B metallo beta-lactamases, which hydrolyze carbapenems and are increasing in prevalence in areas of significant carbapenem usage. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498385 TI - Clinical cancer research 2000: new agents and therapies. PMID- 11498386 TI - Can the heat shock protein 90 inhibitor geldanamycin be designed to specifically inhibit HER-2 tyrosine kinase? PMID- 11498387 TI - Animal use of antimicrobials: impact on resistance. PMID- 11498388 TI - Do Fanconi anemia genes control cell response to cross-linking agents by modulating cytochrome P-450 reductase activity? AB - The Fanconi anemia (FA) genes play an important role in maintaining chromosomal stability and the defense of mammalian cells against cross-linking agents, such as cisplatin and mitomycin C (MMC). Cells derived from FA patients display a characteristic hypersensitivity toward cross-linking agents. Despite great progression in our understanding of the mechanisms that protect cells against these potent anti-cancer drugs, the specific roles of FA gene products in these processes have not been delineated. Recent studies have shown that the FA group C gene product, FANCC, can bind to and regulate the activity of cytochrome P450 reductase (P450R), an enzyme involved in the bioactivation of MMC. In this mini review, this finding is placed in the context of complex mechanisms involved in the bioreductive activation of MMC and the hypersensitivity of FA cells to MMC. Although it would be premature to attribute the FA phenotype wholly to an abnormal activation of MMC, the regulation of P450R by FANCC suggests a novel link between one or more FA gene products, the cellular oxidative state, and the response to chemotherapeutic agents. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498389 TI - Disrupting the p53-mdm2 interaction as a potential therapeutic modality. AB - P53 and mdm2 are linked to each other through a negative feedback loop. P53 transactivates mdm2, but mdm2, in turn, is a major opponent of p53. Mdm2 promotes p53 degradation through a ubiquitin-dependent pathway on 26S proteasomes and is thought to be largely responsible for the very low levels of p53 protein in unstressed cells. The rationale for targeting the p53-mdm2 interaction therapeutically lies in the ability to activate p53 in all those tumors that retain wild type p53. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498390 TI - Anti-angiogenesis therapy:concepts and importance of dosing schedules in clinical trials. AB - The biological control of angiogenesis is critical to the clinical control of cancer. Understanding the mechanism of formation and regulation of new blood vessel development would open a new avenue for cancer treatment. Intense research effort has revealed a variety of factors which initiate, control and terminate the multi-stage process of angiogenesis, as well as target structures which interfere with this process. Protease inhibitors, inhibitors of the endothelial cell proliferation, suppressors of angiogenic growth factors, copper chelators, and other compounds interfering with the process of angiogenesis were screened for inhibition of tumor angiogenesis and some of them are in clinical trials. Very recently, a new term, 'metronomic dosing regimen' has been introduced, which implicates the use of the old cytostatic anticancer agents as anti-angiogenic agents. Results from recent studies will be discussed briefly and the prospects of inhibition of tumor angiogenesis as a new treatment modality will be outlined. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498391 TI - The cost of antibiotic resistance from a bacterial perspective. AB - The emergence, spread and stability of antibiotic resistance in a bacterial population will be determined by several factors including (a) the volume of drug use, (b) the rate of formation of resistant mutants, (c) the biological cost of resistance and (d) the rate and extent of the genetic compensation of the costs. Generally, resistance is associated with a cost, suggesting that the frequency of resistant bacteria might decline when the use of antibiotics is decreased. However, evolution to reduce these costs, without a concomitant loss of resistance, can occur and result in a stabilization of the resistant bacteria in the population. The rate and trajectory of this compensatory evolution is dependent on the bacterial species, the specific resistance mutation and the environmental conditions under which evolution occurs. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498392 TI - Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and impact on treatment. AB - Pseudomonas aeruginosa continues to be a major cause of infections in Western society, in part because of its high intrinsic resistance to antibiotics. It has been demonstrated that this intrinsic resistance arises from the combination of unusually restricted outer-membrane permeability and secondary resistance mechanisms such as energy-dependent multidrug efflux and chromosomally encoded periplasmic beta-lactamase. Given this high level of natural resistance, mutational resistance to most classes of antibiotics can readily arise. In this review we summarize new insights into the mechanisms of resistance, and describe therapeutic approaches that can be used in the face of this continuing resistance threat, as well as new approaches that are being developed to combat resistance. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498393 TI - Is there a role for glyoxalase I inhibitors as antitumor drugs? PMID- 11498394 TI - Infectious disease 2000: drug resistance and new drugs. AB - 40th Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC) was held in Toronto on 17-20 September 2000. It attracted thousands of delegates from industry and academia and covered, in over 2300 oral and poster presentations, topics ranging from microbial pathogenesis to infection control, vaccines, antibiotic resistance and new antimicrobial agents. Summarized here are highlights on microbial resistance and agents in clinical and preclinical development. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498395 TI - cDNA Technologies and their application to drug resistance research: power, potential and problems. AB - The effectiveness of systemic chemotherapy is dramatically limited by both intrinsic and acquired drug resistance. Several new technologies have been developed over the last decade to more rapidly identify underlying genetic alterations that impart a drug-resistant phenotype. Techniques such as cDNA-based subtraction technologies, SAGE analysis, and most recently cDNA array and high density micro-array technologies have rapidly expanded our ability to detect changes in RNA transcription in drug-resistant tumors. These technologies are currently being applied to generate a large number of new hypotheses regarding the regulatory molecules underlying the drug-resistance phenotype. These techniques suggest that there is a large number of transcriptional changes which occur in the drug-resistance phenotype and future work will need to focus on dissecting which of these transcriptional changes are central to the drug resistance phenotype, both in vitro, and more importantly within clinical neoplasia. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498396 TI - Atovaquone resistance in malaria parasites. AB - Atovaquone is a broad-spectrum antiparasitic agent active against malaria, Pneumocystis carinii pneumonia, toxoplasmosis and babesiosis. When used as a single agent, resistance to atovaquone arose rapidly in falciparum malaria, requiring the development of a new antimalarial drug combination of atovaquone and proguanil. Recent laboratory investigations have provided insights into the mode of atovaquone action, and identified the molecular basis for the resistance development. Mutations within a catalytic domain of the cytochrome bc(1)complex present within the parasite mitochondrial inner membrane were shown to be responsible for atovaquone resistance. Here, we review these studies and propose a mechanism by which atovaquone resistance may arise quickly in malaria parasites. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498397 TI - The (patho)physiological functions of the MRP family. AB - The identification of certain members of the large superfamily of ATP binding cassette transport proteins such as MDR1 -P-glycoprotein and the multidrug resistance protein MRP1 as ATP-dependent drug efflux pumps has been a major contribution in our understanding of the multidrug resistance phenotype of cancer cells. Importantly, both transport proteins that exhibit only low structural homology have a very different substrate specificity but confer resistance to a similar spectrum of natural product chemotherapeutic drugs. In contrast to the drug transporter MDR1, MRP1 mainly transports anionic Phase II-conjugates. In addition MRP1-mediated drug resistance is highly dependent on high intracellular glutathione levels which may be linked to the apparent physiological involvement of MRP1 in glutathione-related cellular processes. This review summarizes the current knowledge about functional aspects of MRP1 and its five recently cloned homologues MRP2-MRP6 and discusses their substrate specificities and cellular localization with emphasis on drug resistance. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498398 TI - The impact of antibiotic use on resistance development and persistence. AB - The intense use and misuse of antibiotics are undoubtedly the major forces associated with the high numbers of resistant pathogenic and commensal bacteria worldwide. Both the volume and the way antibiotics are applied contributes to the selection of resistant strains. Still, other social, ecological and genetic factors affect a direct relationship between use and frequency of resistance. Resistant bacteria, following their emergence and evolution in the presence of antibiotics, appear to acquire a 'life of their own'. They proliferate and maintain the resistance traits even in the absence of antibiotics, thus jeopardizing the reversal of bacterial resistance by simple reduction in antibiotic use. Reversing resistance requires restoration of the former susceptible flora in people and in the environment. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498399 TI - Effects of HER-2/neu on chemosensitivity of tumor cells. AB - In solid tumors, such as breast and ovarian cancer, the predominant genetic mechanism for oncogene activation is through gene amplification. The HER-2 (also known as ErbB2/c-erbB2/HER-2 / neu) oncogene is the most frequently amplified oncogene in breast cancer and its overexpression is associated with poor clinical outcome. In addition to its role in tumor progression, HER-2 has been implicated in altering tumor cell chemosensitivity to cytotoxic chemotherapy, particularly to anthracyclines. However, sophisticated in vitro studies have recently indicated that HER-2 may not have anything to do with the sensitivity of the cancer cells to cytotoxic drugs. Topoisomerase IIalpha gene is a target gene for many cytotoxic drugs and is located just by the HER-2 at the 17q12-q21. TopoIIalpha amplification and deletion may account for both relative chemosensitivity and resistance to anthracycline-therapy depending on the specific genetic defect at the topoIIalpha locus. Whereas HER-2 is an oncogene that clearly can drive tumor induction and growth, its function as a marker for chemoselection may be due to associated genetic changes in the topoIIalpha gene. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498400 TI - The role of efflux in macrolide resistance. AB - Efflux is one of the major resistance mechanisms for macrolide antibiotics observed in both laboratory and clinical settings. This review summarizes the recent research on two major macrolide efflux pumps: Mef in Gram-positive organisms and Acr-AB-TolC in Haemophilus influenzae and Escherichia coli. The roles of pumps in macrolide resistance and the new advances / strategies to overcome efflux are discussed. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498401 TI - An ABC-type multidrug transporter of Lactococcus lactis possesses an exceptionally broad substrate specificity. AB - LmrA is a 590-amino acid membrane protein which confers multidrug resistance on Lactococcus lactis cells by extruding amphiphilic compounds from the inner leaflet of the cytoplasmic membrane at the expense of ATP hydrolysis. Its structural and functional characteristics place it in the P-glycoprotein cluster of the ATP-binding cassette transporter superfamily, making it the first prokaryotic multidrug transporter of this cluster. The number of compounds recognized and transported by LmrA is remarkably vast and includes many lipophilic cations as well as a record of eight classes of clinically relevant broad-spectrum antibiotics. Homologs of LmrA have been found in pathogenic bacteria, suggesting that these putative efflux pumps may play a crucial role in antibiotic resistance of human pathogens. Recent evidence indicates that LmrA is functional as a homodimer, consistent with the overall structure of P glycoprotein, and mediates drug transport by an alternating two-site transport mechanism. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498402 TI - The relevance of drug sequence in combination chemotherapy. AB - The concept of combining chemotherapeutic agents to increase the cytotoxic efficacy has evolved greatly over the past several years. In the past, the rationale for combination chemotherapy centered on attacking different biochemical targets, overcoming drug resistance in heterogenous tumors, and increasing the dose-density of combination chemotherapy to take advantage of tumor growth kinetics. The overall goal was to improve clinical efficacy with acceptable clinical toxicity. It is now apparent that the sequence of drug administration can significantly enhance the therapeutic effect of chemotherapy. These sequence-dependent effects can be explained by chemotherapy-induced cell cycle perturbations, or by pharmacodynamic interactions between the agents in combination. In this review, we focus on drug combinations with taxanes and camptothecins, which we believe best illustrate the importance of the cell cycle and pharmacologic interactions in the sequential administration of chemotherapy. As our understanding of the cell cycle grows, our ability to appropriately sequence chemotherapy can have a great impact on the treatment of human cancers. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498403 TI - Does P-glycoprotein play a role in anticancer drug pharmacokinetics? AB - The multidrug-resistance P-glycoprotein is a drug efflux transport protein abundantly present in various types of human cancer. The protein is encoded by the MDR1 gene and its function is sensitive to modulation by competitive inhibition. Clinical studies have indicated that inhibitors of P-glycoprotein function dramatically decrease the systemic clearance of anticancer agents, necessitating dose reduction. This dose reduction not only complicated the interpretation of toxicity and response data, but also presented a serious obstacle in the development and rational use of P-glycoprotein inhibitors. It is now evident that the pharmacokinetic interference between anticancer drugs and P glycoprotein inhibitors is due primarily to competition for drug metabolizing enzymes. A wealth of recent experimental data shows that many of the previously tested P-glycoprotein inhibitors, including verapamil, cyclosporin A, and valspodar (SDZ PSC 833), are substrates and/or potent inhibitors of cytochrome P450 3A4 (CYP3A4). Future development and clinical use of potent P-glycoprotein modulators lacking high affinity for CYP3A4 should decrease the impact of these important drug interactions and will eventually result in improved therapeutic specificity and efficacy. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498404 TI - The role of murine models in the development of antifungal therapy for systemic mycoses. AB - Animal testing is crucial to the development of new antifungal compounds. This review describes the role that murine and other animal models have played in the development of three classes of antifungal agents: the polyenes, the triazoles and the echinocandins and the ways in which these models have been either the positive link in the path from in vitro studies to the patient, or have foreclosed later clinical evaluation. Efficacy studies in particular mycoses are discussed, as well as studies designed to determine whether combinations of antifungal drugs may have value over single agents. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498405 TI - Drug treatment of microsporidiosis. AB - Microsporidia are ubiquitous organisms that are emerging pathogens in humans. These are most likely zoonotic and/or waterborne infections. In the immunosuppressed host, such as those treated with immunosuppressive drugs or infected with human immunodeficiency virus particularly at advanced stages of the disease, microsporidia can produce a wide range of clinical diseases. The most common manifestation is gastrointestinal tract infection; however, encephalitis, ocular infection, sinusitis, myositis and disseminated infection have also been described. In addition, these organisms have been reported in immune competent individuals. Multiple genera are involved in these infections and different organisms can result in distinct clinical pictures. Differences in clinical and parasitologic response to various therapeutic agents have emerged from clinical, as well as in vitro and in vivo studies. Currently there are no precisely defined guidelines for the optimal treatment of microsporidial infections. This article reviews the available data on compounds with in vitro activity and/or in vivo efficacy for microsporidial infections. Copyright 2000 Harcourt Publishers Ltd. PMID- 11498406 TI - Spinal manifestations of intracranial hypotension. PMID- 11498407 TI - Diffusion-weighted imaging as a surrogate marker for stroke as a complication of cerebrovascular procedures and devices. PMID- 11498408 TI - The missing element. PMID- 11498409 TI - Vein of Galen management in neonatal period. PMID- 11498410 TI - Cervical MR imaging in postural headache: MR signs and pathophysiological implications. AB - BACKGROUND AND PURPOSE: Postural headache most often occurs after lumbar puncture as post-lumbar puncture headache (PLPH) or, rarely, spontaneously as spontaneous intracranial hypotension headache (SIHH). In this prospective study, we used spinal MR imaging to determine the findings that would assist in the diagnosis of PLPH and SIHH and that would further our pathophysiological understanding of postural headache. METHODS: The study group consisted of 15 healthy volunteers and 20 patients with postural headache: nine with SIHH and 11 with PLPH. The craniocervical junction and the cervical spine were studied using T2-weighted fast spin-echo and T1-weighted spin-echo sequences in the axial and sagittal planes. Follow-up studies were performed in 13 patients. RESULTS: Dilatation of the anterior internal vertebral venous plexus was the most constant finding, present in 17 (85%) of 20 patients with postural headache. Spinal hygromas, whose location as subdural or epidural could not be exactly determined, were present in 14 patients (70%). A focal fluid collection was detected in the retrospinal region at the C1-C2 level in six patients with SIHH and in four patients with PLPH (50%). Tonsillar descent was detected in only one patient, and subtentorial hygroma in five patients. No abnormalities were found in the volunteers. CONCLUSION: The MR signs of dilatation of the venous plexus, presence of spinal hygromas, and presence of retrospinal fluid collections can help to establish the diagnosis of intracranial hypotension. They are probably the result of decreased CSF volume, with the retrospinal fluid collections being a transudate from the venous plexus rather than frank extravasation. Resolution of these signs parallels resolution of the headache. PMID- 11498411 TI - Diffusion-weighted MR imaging after angioplasty or angioplasty plus stenting of arteries supplying the brain. AB - BACKGROUND AND PURPOSE: There has been concern regarding the safety of revascularization procedures of vessels supplying the brain vessels because of the risk of cerebral embolization during the procedure. We have observed a high incidence of hyperintense lesions on diffusion-weighted MR images of the brain after stenting at the carotid bifurcation. The hypothesis of this study is that diffusion-weighted MR imaging of the brain can reveal new diffusion abnormalities after angioplasty or angioplasty plus stenting of arteries supplying the brain, other than at the carotid bifurcation. Therefore, we prospectively obtained diffusion-weighted MR images of the brain before and after such revascularization procedures. METHODS: Thirty-seven revascularization procedures were performed in 32 patients. Eleven interventions were performed at the distal internal carotid artery, two at the external carotid artery, two at the common carotid artery, five at the innominate artery, five at the vertebral artery, and 12 at the proximal subclavian artery. Diffusion-weighted MR imaging of the brain was performed before and 24 hours after the procedures. RESULTS: After eight (22%) of 37 procedures, new hyperintensities were visible on the diffusion-weighted MR images. With six of these eight procedures, the hyperintensities occurred in the vascular territory supplied by the treated vessel. In total, 35 new cerebral lesions could be seen, 33 (94%) of which occurred in the vascular territory supplied by the treated vessel. None of the patients in whom new diffusion abnormalities were found had new neurologic symptoms or deficits. No new lesions could be seen after procedures at the subclavian artery. CONCLUSION: Revascularization procedures of arteries supplying the brain were associated with new lesions on the diffusion-weighted MR images of the brain after 22% of the procedures, provided that MR imaging could be performed, indicating the occurrence of cerebral microemboli during such procedures. Diffusion-weighted MR imaging of the brain can be used as a tool to assess the impact of modifications of procedural technique and/or the use of cerebral protection devices on the occurrence of such lesions. PMID- 11498412 TI - The value of apparent diffusion coefficient maps in early cerebral ischemia. AB - BACKGROUND AND PURPOSE: Prediction of the regions of the ischemic penumbra that are likely to progress to infarction is of great clinical interest. Whether lowered apparent diffusion coefficient (ADC) values were present in the ischemic penumbra of patients presenting with acute ischemic stroke and were specific to regions of the penumbra that proceeded to infarction was investigated. METHODS: Nineteen patients with hemispheric stroke of less than 6 hours' onset and with acute scans showing a perfusion lesion greater than a diffusion lesion (ischemic penumbra) were studied. Scans also were performed subacutely (days 3 to 5) and at outcome (day 90). The outcome scan was used to identify regions of the penumbra that proceeded to infarction. RESULTS: The ADC ratios were significantly reduced (P <.00001) in regions of the penumbra that progressed to infarction on the outcome scan compared with those that remained normal. In regions that showed transition to infarction, the mean ADC ratios were typically 0.75 to 0.90. CONCLUSION: Intermediate ADC values are present in the ischemic penumbra and are indicative of tissue at risk of infarction. PMID- 11498413 TI - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy: decrease in regional cerebral blood volume in hyperintense subcortical lesions inversely correlates with disability and cognitive performance. AB - BACKGROUND AND PURPOSE: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an arteriopathic syndrome related to a genetic defect on chromosome 19. Characteristic changes in CADASIL can be observed onT2-weighted MR images in the subcortical white matter. The purpose of this study was to measure changes of regional cerebral blood volume (rCBV) with dynamic contrast-enhanced MR imaging and to correlate the changes to disability and cognitive performance. METHODS: We obtained rCBV measurements of 24 individuals with proven CADASIL on a 1.5-T MR imaging unit. A susceptibility weighted MR imaging sequence was used for bolus tracking. Principles of the indicator dilution theory were applied to estimate values of absolute rCBV (mL/100 g). Disability was determined by using the Rankin scale, and overall cognitive performance was assessed by using the Mini-Mental State Examination. RESULTS: The mean rCBV in the subcortical white matter that was hyperintense on the T2-weighted images (2.7 +/- 0.8 mL/100 g) was significantly lower than the rCBV in the white matter that appeared normal on the T2-weighted images (4.4 +/- 1.3 mL/100 g) (P <.05). The mean rCBV in the gray matter was within the normal range (8.3 +/- 1.7 mL/100 g). Both cognitive impairment and disability negatively correlated with rCBV in the subcortical white matter that was hyperintense (P <.05) but not with rCBV in the normal appearing white matter. rCBV did not correlate with age. CONCLUSION: rCBV measured in the hyperintense subcortical white matter in individuals with CADASIL was decreased and inversely correlated with disability and cognitive impairment. PMID- 11498414 TI - Detection of intracranial hemorrhage: comparison between gradient-echo images and b(0) images obtained from diffusion-weighted echo-planar sequences. AB - BACKGROUND AND PURPOSE: Diffusion-weighted MR imaging (DWI) is commonly used as the initial and sole imaging examination for the detection of acute cerebral infarction, yet it remains controversial whether MR can detect hyperacute (<24 h) hemorrhage. Hemorrhage is best detected with gradient-echo (GRE) T2*-weighted sequences, because of their magnetic susceptibility effects. DWI uses a spin-echo echo-planar technique (EPI) that is more sensitive than spin-echo T2-weighted imaging to susceptibility effects. Our aim was to determine whether the b(0) image from the DWI-EPI sequence is as sensitive as GRE in detecting hemorrhagic lesions on imaging studies performed to identify acute infarction or hemorrhage. METHODS: All MR studies performed for clinically suspected or radiographically confirmed acute infarction or hemorrhage from 2/1/98 to 8/15/99 were retrospectively interpreted by one neuroradiologist in a blinded fashion. The sensitivity of hemorrhage detection, conspicuity of lesions, and diagnostic certainty were compared between the b(0) EPI and GRE sequences. RESULTS: We found 101 acute infarcts, of which 13 were hemorrhagic, as evidenced by the presence of hypointensity within the infarction on the GRE sequence. This finding served as the reference standard for detection of hemorrhage. Hemorrhage was diagnosed with confidence in only seven cases (54%) on b(0) images; 22 acute hematomas were hypointense on GRE images whereas 19 were hypointense on b(0) images (86%); 17 chronic hematomas were depicted on GRE images and 12 on b(0) scans (63%). Punctate hemorrhages and linear cortical staining were detected on 37 GRE studies but on only four b(0) studies. Hemorrhage was always more conspicuous on the GRE sequences. CONCLUSION: b(0) images from a DWI sequence failed to detect minimally hemorrhagic infarctions and small chronic hemorrhages associated with microangiopathy. GRE scans were more sensitive than b(0) images in the detection of these hemorrhages and should be included in emergency brain MR studies for acute infarction, especially when thrombolytic therapy is contemplated. PMID- 11498415 TI - MR imaging of the brain in lymphomatoid granulomatosis. AB - BACKGROUND AND PURPOSE: Clinical diagnosis of lymphomatoid granulomatosis (LG) of the brain, in patients without skin or chest lesions, usually is difficult because of the nonspecific neurologic manifestations, laboratory data, and CT appearance. Our aim was to characterize the MR appearance of LG of the brain. METHODS: We retrospectively reviewed the MR images in four patients (35 to 72 years old) with histologically confirmed LG of the brain. RESULTS: On T2-weighted images, we noted diffuse hyperintense lesions in the cerebral white matter bilaterally (n = 3), in the brain stem and cerebellar hemisphere (n = 1), and patchy hyperintense lesions the brain stem (n = 2). On contrast-enhanced T1 weighted images, we observed multiple punctate or linear enhancements residing along the perivascular space (n = 4), nodular enhancements (n = 2), ringlike enhancements (n = 1), and a large, enhanced mass (n = 1). All patients had multifocal lesions. CONCLUSION: Although the MR appearance of LG of the brain varies, multiple punctate or linear enhancements that reside along the perivascular space suggest LG. PMID- 11498416 TI - (18)F-fluorodeoxyglucose positron emission tomography and MR imaging findings in Rasmussen encephalitis. AB - BACKGROUND AND PURPOSE: Rasmussen encephalitis is a chronic, progressive encephalitis that manifests as an abrupt-onset, intractable seizure disorder in previously developmentally normal children. The objectives of the current study were to characterize the (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) and MR imaging findings in Rasmussen encephalitis and to test the hypotheses that data from both imaging techniques are required to establish the diagnosis and identify the affected cerebral hemisphere in some cases. METHODS: Eleven patients with Rasmussen encephalitis were identified from a review of a computer database. The MR (n = 10) and PET (n = 11) imaging data were reviewed retrospectively and conjointly. RESULTS: On MR images, nine of 10 patients manifested bilateral cerebral atrophy that predominantly involved one hemisphere. One patient had purely unilateral cerebral atrophy. We observed foci of abnormally increased T2 signal intensity in nine of 10 patients. On FDG PET images, all patients showed extensive regions of hypometabolism within the cerebral hemisphere that showed the greatest atrophy. Discrete foci of hypermetabolism, indicative of seizure activity, were observed in six patients. The FDG PET and MR imaging findings were either stable or gradually progressive in patients with multiple imaging studies (MR, n = 5; FDG PET, n = 5). CONCLUSION: Rasmussen encephalitis is characterized by diffuse, unilateral cerebral hypometabolism on FDG PET images, with corresponding regions of cerebral atrophy on MR images. Although MR imaging data alone are sufficient to suggest a diagnosis of Rasmussen encephalitis in many cases, correlation with FDG PET data increases diagnostic confidence and allows the unequivocal identification of the affected cerebral hemisphere in patients whose MR imaging findings are subtle or distributed bilaterally. PMID- 11498417 TI - Findings in a patient with herpes simplex viral meningitis associated with acute retinal necrosis syndrome. AB - We report the case of a 41-year-old man with clinical findings of viral meningitis associated with acute retinal necrosis syndrome in his right eye. MR images showed right optic nerve enlargement and high-intensity signal abnormalities in the region of the left lateral geniculate body and the left occipital lobe. PMID- 11498418 TI - CT and MR findings of anthrax meningoencephalitis: report of two cases and review of the literature. AB - Anthrax meningoencephalitis is a rare complication of infection with Bacillus anthracis and generally produces a hemorrhagic meningoencephalitis. We present the CT and MR imaging findings in two patients demonstrating subarachnoid, intracerebral, and intraventricular hemorrhage with leptomeningeal enhancement. PMID- 11498419 TI - Perfusion-sensitive MR imaging of gliomas: comparison between gradient-echo and spin-echo echo-planar imaging techniques. AB - BACKGROUND AND PURPOSE: The different sensitivities to vessel size of gradient echo echo-planar imaging (GE-EPI) and spin-echo EPI (SE-EPI) might indicate the relative cerebral blood volumes (rCBVs) of different tumor sizes. The techniques of GE-EPI and SE-EPI were compared for detecting low- versus high-grade gliomas. METHODS: Six patients with low-grade gliomas and 19 patients with high-grade gliomas underwent two perfusion-sensitive MR procedures, one produced by a GE- and the other by an SE-EPI technique. Maximum rCBV ratios normalized with rCBV of contralateral white matter were calculated for evaluation. P <.05 was considered statistically significant. RESULTS: Maximum rCBV ratios of high-grade gliomas obtained with the GE-EPI technique (mean, 5.0 +/- 2.9) were significantly higher than those obtained with the SE-EPI technique (mean, 2.9 +/- 2.3) (P =.02). Maximum rCBV ratios of low-grade gliomas obtained with the GE-EPI technique (mean, 1.2 +/- 0.7) were almost equal to those obtained with the SE-EPI technique (mean, 1.2 +/- 0.6), and there was no significant difference (P =.66). The difference in the maximum rCBV ratios between the low- and high-grade gliomas reached significance when obtained with the GE-EPI technique (P =.01). CONCLUSION: The GE-EPI technique seems more useful for detecting low- versus high grade gliomas than the SE-EPI technique. PMID- 11498420 TI - Proton MR spectroscopic evaluation of suspicious brain lesions after stereotactic radiotherapy. AB - BACKGROUND AND PURPOSE: The radiologic assessment of suspicious brain lesions after stereotactic radiotherapy of brain tumors is difficult. The purpose of our study was to define parameters from single-voxel proton MR spectroscopy that provide a probability measure for differentiating neoplastic from radiation induced, nonneoplastic lesions. METHODS: Seventy-two lesions in 56 patients were examined using a combined MR imaging and MR spectroscopy protocol (point-resolved spectroscopy, TE = 135 ms). Signal intensities of cholines, creatines, N-acetyl aspartate, and the presence of lactate and lipid resonances were correlated to final diagnoses established by clinical and MR imaging follow-up, positron emission tomography studies, or biopsy/surgery. Statistical analysis was performed using the t test, linear discriminant analysis, and k nearest-neighbor method. RESULTS: Significantly increased signal intensity ratios I(tCho)/I(tCr) (P <.0001) and I(tCho)/I(NAA) (P <.0001) were observed in neoplastic (n = 34) compared with nonneoplastic lesions (n = 32) and contralateral normal brain (n = 33). Analysis of I(tCho)/I(tCr) and I(tCho)/I(NAA) data yielded correct retrospective classification as neoplastic and nonneoplastic in 82% and 81% of the lesions, respectively. Neither I(NAA)/I(tCr) nor signal intensitities of lactate or lipids were useful for differential diagnosis. CONCLUSION: Metabolic information provided by proton MR spectroscopy is useful for the differentiation of neoplastic and nonneoplastic brain lesions after stereotactic radiotherapy of brain tumors. PMID- 11498421 TI - Frequencies contributing to functional connectivity in the cerebral cortex in "resting-state" data. AB - BACKGROUND AND PURPOSE: In subjects performing no specific cognitive task ("resting state"), time courses of voxels within functionally connected regions of the brain have high cross-correlation coefficients ("functional connectivity"). The purpose of this study was to measure the contributions of low frequencies and physiological noise to cross-correlation maps. METHODS: In four healthy volunteers, task-activation functional MR imaging and resting-state data were acquired. We obtained four contiguous slice locations in the "resting state" with a high sampling rate. Regions of interest consisting of four contiguous voxels were selected. The correlation coefficient for the averaged time course and every other voxel in the four slices was calculated and separated into its component frequency contributions. We calculated the relative amounts of the spectrum that were in the low-frequency (0 to 0.1 Hz), the respiratory-frequency (0.1 to 0.5 Hz), and cardiac-frequency range (0.6 to 1.2 Hz). RESULTS: For each volunteer, resting-state maps that resembled task-activation maps were obtained. For the auditory and visual cortices, the correlation coefficient depended almost exclusively on low frequencies (<0.1 Hz). For all cortical regions studied, low frequency fluctuations contributed more than 90% of the correlation coefficient. Physiological (respiratory and cardiac) noise sources contributed less than 10% to any functional connectivity MR imaging map. In blood vessels and cerebrospinal fluid, physiological noise contributed more to the correlation coefficient. CONCLUSION: Functional connectivity in the auditory, visual, and sensorimotor cortices is characterized predominantly by frequencies slower than those in the cardiac and respiratory cycles. In functionally connected regions, these low frequencies are characterized by a high degree of temporal coherence. PMID- 11498422 TI - Detection of mesial temporal lobe hypoperfusion in patients with temporal lobe epilepsy by use of arterial spin labeled perfusion MR imaging. AB - BACKGROUND AND PURPOSE: Interictal hypometabolism has lateralizing value in cases of temporal lobe epilepsy and positive predictive value for seizure-free outcome after surgery to treat epilepsy. Alterations in regional cerebral metabolism can also be inferred from measurements of regional cerebral perfusion. The purpose of this study was to determine the feasibility of detecting cerebral blood flow (CBF) asymmetries in the mesial temporal lobes using continuous arterial spin labeling perfusion MR imaging, which is a noninvasive method for calculating regional CBF. METHODS: Twelve patients with medically refractory temporal lobe epilepsy who underwent preoperative evaluation for temporal lobectomy and 12 normal control participants were studied retrospectively. Absolute and normalized mesial temporal CBF measurements were compared between the patient and control groups. Lateralization based on a perfusion asymmetry index was compared with metabolic ((18)[F]-fluorodeoxyglucose positron emission tomography) and hippocampal volumetric asymmetry indices and with clinical lateralization. RESULTS: Mesial temporal CBF was more asymmetric in patients with temporal lobe epilepsy than in normal control participants, although asymmetric mesial temporal CBF was also found in normal participants, with the left side dominant. Ipsilateral mesial temporal CBF was significantly decreased compared with contralateral mesial temporal CBF in patients with temporal lobe epilepsy. Global CBF measurements were significantly decreased in patients compared with control participants. Asymmetry in mesial temporal blood flow in patients persisted after normalization to global CBF. Lateralization using continuous arterial spin labeling perfusion MR imaging asymmetry index significantly correlated with lateralization based on (18)[F]-fluorodeoxyglucose positron emission tomography hypometabolism, hippocampal volumes, and clinical evaluation. CONCLUSION: Continuous arterial spin labeling perfusion MR imaging can detect interictal asymmetries in mesial temporal lobe perfusion in patients with temporal lobe epilepsy. This technique is readily combined with routine structural assessment and potentially offers an inexpensive and noninvasive means of screening for asymmetries in interictal mesial temporal lobe function. PMID- 11498423 TI - Amygdalar and hippocampal volumetry in control participants: differences regarding handedness. AB - BACKGROUND AND PURPOSE: Amygdalar and hippocampal volume measurements indicate a right-greater-than-left asymmetry for right-handed normal participants in most studies. The purpose of this study was to compare amygdalar and hippocampal volume ratios between right- and left-handed participants. METHODS: Amygdalar and hippocampal volume measurements were performed in 34 (20 right-handed and 14 left handed) normal participants. All participants completed a 10-item handedness questionnaire. The MR imaging sequence was a 3D T1-weighted gradient-echo acquisition of the whole brain (24/6 [TR/TE]; flip angle, 25 degrees). MR images were spatially normalized, and volumes were painted with a 1.0-mm(3) resolution cursor on an SGI workstation. RESULTS: In right-handed participants, the amygdala and hippocampus (P <.001 for both) were significantly larger on the right side than on the left. The left-handed group did not show a significant difference between right- and left-sided structures. Right-to-left volume ratios differed significantly between right- and left-handed participants for both amygdalar (P <.02) and hippocampal (P <.01) structures. Gender did not affect right-to-left amygdalar and hippocampal volume ratios in right- or left-handed participants. CONCLUSION: Handedness, but not gender, seems to affect right-to-left amygdalar and hippocampal volume ratios. PMID- 11498424 TI - De novo aneurysm formation after stereotactic radiosurgery of a residual arteriovenous malformation: case report. AB - We report a case of a 19-year-old woman who underwent radiosurgical treatment of a residual arteriovenous malformation. Nine months after treatment, repeat angiography revealed a de novo paranidal aneurysm that was treated endovascularly. We postulate that changes in flow dynamics or vessel integrity after radiosurgery contributed to the formation of her de novo aneurysm. PMID- 11498425 TI - MR diagnosis of myxomatous aneurysms: report of two cases. AB - We report two cases in which MR imaging was used to diagnose multiple cerebral aneurysms caused by cardiac myxoma. In one case, the diagnosis was confirmed with cerebral angiography. Myxomatous aneurysms characteristically appeared on T1 weighted images as contrast-enhancing focal fusiform dilatations of distal segments of the cerebral arteries. On T2-weighted images, these myxomatous aneurysms appeared as low signal intensity flow voids, sometimes associated with cerebral infarctions. PMID- 11498426 TI - Imaging of cerebral and brain stem amyloidomas. AB - CNS amyloidomas are rare. We describe a 51-year-old man with isolated amyloidomas in the cerebral white matter and in the pons. CT and MR imaging showed a heterogeneous, enhancing mass in the deep cerebral white matter. A second, much smaller linear serpiginous lesion was present in the pons. PMID- 11498427 TI - Persistence of gadolinium in CSF: a diagnostic pitfall in patients with end-stage renal disease. AB - Two dialysis-dependent patients with end-stage renal disease underwent brain and spine MR imaging a few days after having undergone gadolinium-enhanced imaging studies. Increased signal intensity in the subarachnoid space on T1-weighted and fluid-attenuated inversion recovery images was noted. Excretion of gadolinium into the CSF was proven in one case by mass spectrometry. Dialysis-dependent patients with end-stage renal disease and neurologic abnormalities often undergo contrast-enhanced MR imaging. Recognition that these patients may show increased signal intensity in the subarachnoid space because of gadolinium excretion into CSF may prevent diagnostic errors. PMID- 11498428 TI - Balo's concentric sclerosis: clinical and radiologic features of five cases. AB - Balo's concentric sclerosis (BCS) is a rare demyelinating disease considered to be a variant of multiple sclerosis. Five BCS cases were diagnosed antemortem based on their typical concentric mass patterns on MR images and based on clinical and CSF findings. Histopathologic investigation was also performed in one case. Our case report supports the concept that BCS may be a self-limited disease that is not always fatal. Characteristic MR imaging findings may allow antemortem diagnosis of BCS when performed at the onset of the disease. PMID- 11498429 TI - Dumbbell schwannomas of the internal auditory canal. AB - BACKGROUND AND PURPOSE: Benign tumors of the internal auditory canal (IAC) may leave the confines of the IAC fundus and extend into inner ear structures, forming a dumbbell-shaped lesion. It is important to differentiate dumbbell lesions, which include facial and vestibulocochlear schwannomas, from simple intracanalicular schwannomas, as surgical techniques and prognostic implications are affected. In this article, the imaging and clinical features of these dumbbell schwannomas are described. METHODS: A dumbbell lesion of the IAC is defined as a mass with two bulbous segments, one in the IAC fundus and the other in the membranous labyrinth of the inner ear or the geniculate ganglion of the facial nerve canal, spanned by an isthmus. Twenty-four patients with dumbbell lesions of the IAC had their clinical and imaging data retrospectively reviewed. Images were evaluated for contour of the mass and extension into the membranous labyrinth or geniculate ganglion. RESULTS: Ten of 24 lesions were facial nerve dumbbell lesions. Characteristic features included an enhancing "tail" along the labyrinthine segment of the facial nerve and enlargement of the facial nerve canal. Dumbbell schwannomas of the vestibulocochlear nerve (14/24) included transmodiolar (8/14), which extended into the cochlea, transmacular (2/14), which extended into the vestibule, and combined transmodiolar/transmacular (4/14) types. CONCLUSION: Simple intracanalicular schwannomas can be differentiated from transmodiolar, transmacular, and facial nerve schwannomas with postcontrast and high-resolution fast spin-echo T2-weighted MR imaging. Temporal bone CT is reserved for presurgical planning in the dumbbell facial nerve schwannoma group. PMID- 11498430 TI - Cerebellopontine angle ganglioglioma: MR findings. AB - We present a case of cerebellopontine (CP) angle ganglioglioma in a young child with developmental delay and no trigeminal nerve symptoms. MR imaging demonstrated a mass of homogeneous low signal intensity in the left CP angle on T1-weighted images with no enhancement with gadolinium, and of relatively homogeneous high signal intensity on T2-weighted images. PMID- 11498431 TI - Enhancement of the eighth cranial nerve and labyrinth on MR imaging in sudden sensorineural hearing loss associated with human herpesvirus 1 infection: case report. AB - The case of a 61-year-old woman who presented with herpes labialis, subclinical meningitis, and sudden onset of bilateral sensorineural hearing loss is presented. Contrast-enhanced MR imaging showed marked bilateral enhancement of the intracanalicular portion of the eighth cranial nerve, right cochlea, and left vestibule. Polymerase chain reaction was positive for human herpesvirus 1 obtained from the cerebral spinal fluid, which suggested the diagnosis of viral neuritis. PMID- 11498432 TI - Intraosseous hemangioma of the zygoma: CT and MR findings. AB - Intraosseous hemangiomas are uncommon, constituting less than 1% of all osseous tumors. The most frequent sites are the calvaria and the vertebral column. Involvement of the facial bones is rare, and occurs most commonly in the maxilla, mandible, and nasal bones. Only 20 cases of zygomatic involvement have been reported in the English-language literature. We report a case of an intraosseous hemangioma of the zygoma documented by CT and MR studies. PMID- 11498433 TI - Rosai-Dorfman disease presenting with isolated bilateral orbital masses: report of two cases. AB - Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) is a rare benign idiopathic proliferative disease of phagocytic histiocytes. Approximately 80% of patients present with painless massive cervical lymphadenopathy. Isolated extranodal involvement is relatively uncommon. Two cases of Rosai-Dorfman disease are reported: one with isolated bilateral orbital involvement and one with marked cervical lymphadenopathy and multiple dural-based and intraventricular masses. PMID- 11498434 TI - Merkel cell carcinoma: a rare cause of hypervascular nasal tumor. AB - Cutaneous neuroendocrine carcinoma, first described in 1972, is an aggressive disease usually occurring in sun-exposed skin. Other sites have been described, however; such tumors occasionally occur within the nasal fossa. A high rate of metastasis (>30%) explains the poor prognosis. Descriptions of the imaging features of these tumors, mainly located in cutaneous region, are rare. We therefore present the imaging features of two cases of Merkel cell carcinoma involving the sinonasal region, suggestive of a hypervascular tumor. PMID- 11498435 TI - Unusual CT and MR findings of inflammatory pseudotumor in the parapharyngeal space: case report. AB - Unusual MR and CT findings of an inflammatory pseudotumor in the parapharyngeal space of a 73-year-old woman are reported. The mass was hypointense on T1- and T2 weighted images and demonstrated ring enhancement after contrast medium injection. Punctated calcifications were scattered at the periphery. Inflammatory pseudotumors in the parapharyngeal space are rare, and only three cases have been reported. The possible pathogenesis and varieties of inflammatory pseudotumors are discussed. PMID- 11498436 TI - Cutaneous emphysema and craniocervical bone pneumatization. AB - We report a case of pneumatization of the upper cervical spine and the craniocervical junction, including the occipital bone, accompanied by extensive soft tissue emphysema. There was no history of trauma or surgery. Follow-up X-ray and CT demonstrated the development of those changes. A combination of a developmental abnormality and the unusual habit of frequent Valsalva's maneuvers may have led to those findings. Clinical consequences will be discussed. PMID- 11498437 TI - CT findings associated with Eagle syndrome. AB - Eagle syndrome is an aggregate of symptoms caused by an elongated ossified styloid process, the cause of which remains unclear. This is a rare finding that often goes undetected in the absence of radiographic studies. In this case, we present the diagnostic CT and lateral view plain film radiography findings of a 39-year-old woman with clinical evidence of Eagle syndrome. Eagle syndrome can occur unilaterally or bilaterally and most frequently results in symptoms of dysphagia, headache, pain on rotation of the neck, pain on extension of the tongue, change in voice, and a sensation of hypersalivation (1, 2). We present rare and diagnostic radiographic evidence of this on both plain film radiographs and CT scans. Although well documented in otolaryngology literature and dentistry literature, this syndrome has not been reported in the radiology literature. PMID- 11498438 TI - Endovascular management of vein of Galen aneurysmal malformations presenting in the neonatal period. AB - BACKGROUND AND PURPOSE: Neonates with vein of Galen aneurysmal malformations (VGAMs) presenting with cardiac failure have high morbidity and mortality, and outcomes are significantly better in those presenting in later childhood. Neurologic outcomes in survivors are perceived to be uniformly poor, which may lead to the neonate being denied treatment. We assessed outcomes of modern neonatal intensive care and endovascular embolization in a consecutive series of such neonates presenting with cardiac failure. METHODS: Between 1996 and 1998, five infants (three male, two female) were diagnosed with symptomatic VGAMs in the first week of life, four of whom had intractable, high-output cardiac failure and underwent initial endovascular treatment. There were 15 endovascular procedures and one neurosurgical clipping in these five patients. Transarterial and transvenous routes were required, using multiple embolic agents. We emphasized the use of sonographically guided, percutaneous transtorcular-venous access, moveable-core guidewire as an embolic agent; routine MR imaging; and MR angiography. RESULTS: Immediate outcomes included control of cardiac failure with normal neurologic function in four (80%) patients and one (20%) death from intractable cardiac failure. On follow-up examination, three (60%) infants showed no evidence of neurologic abnormality or cardiac failure; one (20%) infant showed moderate developmental delay. Two have had no further shunting on angiography, one has minimal flow, and one is awaiting follow-up imaging. CONCLUSION: Endovascular therapy with modern neuroanesthetic and neurointensive care can provide good outcomes even in the highest-risk neonates with VGAMs and cardiac failure. If medical management of cardiac failure fails, and there is no evidence of gross cerebral parenchymal damage on imaging, urgent endovascular treatment is feasible and can reduce the almost-100% mortality otherwise expected, without invariably severe morbidity. Use of multiple embolization strategies in multiple stages usually is necessary in these patients, and novel approaches and embolic agents may be necessary. PMID- 11498439 TI - Delivery of human vascular endothelial growth factor with platinum coils enhances wall thickening and coil impregnation in a rat aneurysm model. AB - BACKGROUND AND PURPOSE: Histopathologic studies indicate that aneurysms treated with Guglielmi detachable coils (GDCs) have avascular centers with fibrosis mostly at the aneurysm periphery. We hypothesized that vascular endothelial growth factor (VEGF) released from a coil promotes clot organization, hyperplasia, and endothelial proliferation to facilitate closure of the aneurysm neck. METHODS: GDC segments were inserted into ligated common carotid arteries (CCAs) of adult male rats for 14 days. Coil segments (4-mm) were unmodified, modified with type I collagen (2.4 mg/mL), or modified with type I collagen and recombinant human VEGF-165 (rhVEGF; 500 microg/mL). CCA segments were harvested and coils removed for scanning electron microscopy (SEM). RESULTS: Collagen/rhVEGF coils (n = 11) resulted in marked reductions in CCA lumen area (0.03 mm(2)) compared with coils (n = 9, 0.21 mm(2), P <.001) and collagen coils (n = 5, 0.13 mm(2), P <.001). Collagen/rhVEGF coils (n = 11) also resulted in marked reductions in CCA diameter (0.19 mm) compared with coils (n = 9, 0.50 mm, P <.001) and collagen coils (n = 5, 0.40 mm, P <.001). Wall thickness was greater for the collagen/rhVEGF coil segments (0.22 mm) compared with coils (0.09 mm, P <.001), and the collagen coils (0.15 mm, P =.06). CCA segments containing collagen/rhVEGF coils also displayed Factor VIII positivity and were completely encapsulated in fibrotic tissue, while the unmodified and collagen coils were essentially smooth, as seen by SEM. CONCLUSION: These results suggest that rhVEGF may be beneficial in promoting endothelialization, clot organization, and tissue integration of the coils. This is the first study to hypothesize that rhVEGF may be useful as a surface modification to GDCs for enhancing their therapeutic effects in the treatment of cerebral aneurysms. PMID- 11498440 TI - Treatment of a giant orbitofacial vascular malformation and ophthalmic artery aneurysms with intralesion glue injections: case report. AB - We describe a giant left orbital vascular malformation that was treated with both percutaneous and transarterial embolization. Feeder artery aneurysms thrombosed as a result of retrograde reflux of embolic material into the distal ophthalmic artery. In this presentation, we emphasize the efficacy of percutaneous embolization and retrograde thromboses of two intradural saccular ophthalmic artery aneurysms. PMID- 11498442 TI - Unilateral aplasia of the middle cranial fossa floor in atypical hemifacial microsomia. AB - We present the first report of a patient with atypical hemifacial microsomia (HFM) and unilateral aplasia of the floor of the middle cranial fossa, glenoid fossa, and portions of her posterior fossa. The patient also developed a Chiari I malformation with cervical syrinx over a 3-year interval. This case report highlights the critical role of imaging in revealing serious, but clinically occult, structural abnormalities, as well as the evolution in the pathogenetic understanding of HFM. PMID- 11498441 TI - Three-dimensional proton MR spectroscopic imaging of premature and term neonates. AB - BACKGROUND AND PURPOSE: Previous studies have primarily used single-voxel techniques to obtain MR spectra from the neonatal brain. In this study, we applied 3D MR spectroscopic imaging techniques to detect the spatial distribution of MR spectroscopic imaging-detectable compounds in premature and term infants. The goals were to test the feasibility of obtaining 3D MR spectroscopic images of newborns, assess the spatial variations of metabolite levels, and determine age dependent differences in MR spectroscopic imaging data. METHODS: MR spectroscopic imaging data were acquired from nine premature (postconceptional age, 30-34 weeks) and eight term (postconceptional age, 38-42 weeks) neonates, all with normal clinical and neurologic outcomes. A specialized point-resolved spectroscopy sequence with very selective saturation pulses was used to select a region encompassing the majority of the brain. Phase encoding in three dimensions was performed in a 17-minute acquisition time to obtain 3D spectral arrays with a 1.0 cm(3) nominal spatial resolution. RESULTS: This study showed the feasibility of detecting the 3D distributions of choline, creatine, and N-acetylaspartate resonances in the neonatal brain. Significant spectral differences were detected among anatomic locations and between the premature and term groups. CONCLUSION: This initial study indicates that 3D MR spectroscopic imaging of the neonatal brain can detect anatomic and age-dependent variations in metabolite levels. This technique seems to be a powerful tool to assess the metabolic differences between anatomic regions and to follow the changes in cellular metabolites with brain maturation. This study also indicates the need for determining topologic and age matched normative values before metabolic abnormalities in neonates can be accurately assessed by MR spectroscopy. PMID- 11498445 TI - Contrast enhancement of a hyperacute spontaneous spinal epidural hematoma. PMID- 11498446 TI - A time to stop prescribing antioxidant vitamins to prevent and treat heart disease? PMID- 11498447 TI - The first human monoclonal antibody to oxidized LDL. PMID- 11498448 TI - Statins inhibit leukocyte recruitment: new evidence for their anti-inflammatory properties. PMID- 11498449 TI - Inducible NO synthesis in the vasculature: molecular context defines physiological response. PMID- 11498450 TI - Is the emperor wearing clothes? Clinical trials of vitamin E and the LDL oxidation hypothesis. AB - A wealth of evidence indicates that oxidized low density lipoprotein (LDL) may be of central importance in animal models of atherogenesis. In recent clinical trials, however, dietary vitamin E supplements have not consistently prevented cardiac events in humans with established coronary artery disease. Such mixed results have led many to question the role of LDL oxidation in human atherosclerosis, although this interpretation assumes that the doses of vitamin E used in the studies inhibited lipid oxidation in vivo. In fact, there is remarkably little evidence indicating that those particular regimens effectively inhibit lipid peroxidation in healthy humans. Moreover, evidence of increased oxidative stress was not a criterion for inclusion in the trials; therefore, vitamin E may have benefited only a subset of the participants. These uncertainties raise doubts about the ability of vitamin E to augment antioxidant defense mechanisms in vivo and leave many questions about LDL oxidation and atherosclerosis unanswered. PMID- 11498451 TI - Raloxifene-mediated increase in matrix metalloproteinase-1 production by activated monocytes. AB - Matrix metalloproteinases (MMPs), proteolytic enzymes produced by monocytes, may contribute to atherosclerotic arterial wall remodeling and to plaque rupture. Because estrogen influences the synthesis of MMPs, we examined the effect of raloxifene, a selective estrogen receptor modulator, on monocyte MMP production. Human primary blood monocytes treated with raloxifene (10 micromol/L) in the presence of lipopolysaccharide (LPS) or tumor necrosis factor-alpha and granulocyte-macrophage colony-stimulating factor induced a 2- to 3-fold increase in MMP-1 production by monocytes. The enhancement of MMP-1 production by raloxifene in LPS-activated monocytes occurred through a cyclooxygenase-2- and prostaglandin E(2)-independent mechanism. Additionally, compared with monocytes acquired during the placebo phase, peripheral blood monocytes from 5 of 6 healthy postmenopausal women treated with raloxifene (60 mg daily for 1 month) in a clinical trial produced significantly higher levels of MMP-1 when the monocytes were activated with LPS. Furthermore, serum obtained during the raloxifene phase from 4 of these subjects, when added to control monocytes, significantly enhanced LPS-induced MMP-1 production compared with that from serum obtained during the placebo phase. In summary, raloxifene increases the production of MMP-1 in activated monocytes; this effect may be favorable in atherosclerotic arterial wall remodeling but unfavorable for plaque stability. PMID- 11498452 TI - Arterial elastase activity after balloon angioplasty and effects of elafin, an elastase inhibitor. AB - Increased proteolytic activity may be a factor in intimal hyperplasia after balloon angioplasty (BA). The objectives of this study were to assess elastase activity after BA in a rabbit arterial double-injury model and the effects of elastase inhibition. Elastase activity increased immediately after BA, reached an 8-fold peak at 1 week, and declined to baseline levels by 4 weeks. Elastin zymography showed that the elastase activity was associated predominantly with a molecular mass of 25 kDa. Elastase activity was significantly inhibited in vitro by elafin and phenylmethylsulfonyl fluoride, selective inhibitors of serine elastases. A second group of animals was transfected after BA with a plasmid containing the cDNA for either elafin or a control (chloramphenicol acetyltransferase, CAT) construct by using a hemagglutinating virus of Japan liposome transfection technique. Arterial segments were obtained at 48 hours, 1 week, and 4 weeks to assess transgene expression, arterial wall elastase activity, and intimal cross-sectional area, respectively. Elafin transgene expression was evident at 48 hours and resulted in a significant (80%) inhibition of elastase activity compared with chloramphenicol acetyltransferase-transfected arteries. There was a 43% reduction in intimal cross-sectional area in elafin transfected arteries (0.28+/-0.22 versus 0.16+/-0.07 mm(2) for CAT-transfected versus elafin-transfected arteries, respectively; P<0.05). These data suggest that an early increase in serine elastase activity after BA contributes to intimal hyperplasia. Serine elastase inhibition may be a potential therapeutic approach to inhibit intimal hyperplasia. PMID- 11498453 TI - Local gene transfer of tissue inhibitor of metalloproteinase-2 influences vein graft remodeling in a mouse model. AB - Recently, we established a new mouse model of vein graft arteriosclerosis by grafting vena cava to carotid arteries. In many respects, the morphological features of this murine vascular graft model resemble those of human venous bypass graft disease. Using this model, we studied the effects of local gene transfer of tissue inhibitor of metalloproteinase-2 (TIMP-2) on vein graft remodeling. Mouse isogeneic vessels of the vena caval veins were grafted end to end into carotid arteries, then enveloped with the replication-defective recombinant adenoviruses overexpressing human TIMP-2 (RAdTIMP-2) or beta galactosidase (RAdLacZ) at 1x10(10) plaque-forming units/mL in a total volume of 50 microL, and incubated at room temperature for 20 minutes. In the untreated group, vessel wall thickening was observed as early as 1 week after surgery and progressed to 4- to 10-fold the original thickness in grafted veins at 4 and 8 weeks, respectively. RAdLacZ vector treatment significantly enhanced neointimal lesions at 8 weeks, which was completely blocked by RAdTIMP-2 gene overexpression. Interestingly, RAdTIMP-2 gene transfer resulted in a reduction in vessel diameter of grafted veins compared with ungrafted veins (819+/-96 versus 624+/-67 microm, respectively; P<0.05). Maximal beta-galactosidase activity was found at 2 weeks and was detectable until 4 weeks after gene transfer. Double immunofluorescence studies demonstrated that cells overexpressing TIMP-2 were mostly localized in the adventitia and were MAC-1-positive monocytes/macrophages but not smooth muscle cells. Furthermore, the activity of matrix metalloproteinases was markedly decreased in the vessel walls treated with RAdTIMP-2 compared with that in the untreated control group and the RAdLacZ treated group. Thus, this mouse model has been proven to be useful in gene transfer studies. Our findings demonstrate that local TIMP-2 gene transfer significantly reduces vein graft diameter, ie, remodeling to an artery-like vessel via inhibition of matrix metalloproteinase activity. PMID- 11498454 TI - NO-dependent vasorelaxation is impaired after gene transfer of inducible NO synthase. AB - Proinflammatory stimuli produce expression of inducible NO-synthase (iNOS) within blood vessels and are associated with impaired endothelium-dependent relaxation. Gene transfer of iNOS was used to test the hypothesis that expression of iNOS in blood vessels produces impairment of NO-dependent relaxation as well as contraction. An adenoviral vector containing cDNA for murine iNOS, AdCMViNOS, and a control virus, AdCMVBglII, were used for gene transfer to rabbit carotid arteries in vitro and in vivo. After gene transfer of iNOS in vitro, contractile responses to KCl, phenylephrine, and U46619 were impaired. Relaxation in response to acetylcholine, ADP, A23187, and nitroprusside was also impaired. For example, maximum relaxation of vessels to acetylcholine (10 micromol/L) was 78+/-4% (mean+/-SE) after AdBglII (10(10.5) plaque-forming units) and 34+/-5% after AdiNOS (10(10.5) plaque-forming units, P<0.05). NO-independent relaxation in response to 8-bromo-cGMP and papaverine was not impaired after AdiNOS. Contraction and relaxation were improved in carotid arteries expressing iNOS by aminoguanidine and L-N-iminoethyl lysine, inhibitors of iNOS. After intraluminal gene transfer of iNOS in vivo, contraction of vessels in vitro was normal, but responses to acetylcholine were impaired. In summary, the major finding is that NO-dependent relaxation is impaired in arteries after gene transfer of iNOS in vitro and in vivo. Thus, expression of iNOS per se impairs NO-dependent relaxation. PMID- 11498455 TI - Involvement of Rho-kinase and the actin filament network in angiotensin II induced contraction and extracellular signal-regulated kinase activity in intact rat mesenteric resistance arteries. AB - We have previously shown that angiotensin II (Ang II) and pressure increase extracellular signal-regulated kinase (ERK) 1/2 activity synergistically in intact, pressurized resistance arteries in vitro. However, the mechanisms by which pressure and Ang II activate ERK1/2 in intact resistance arteries remain to be determined. The purpose of the present study was to investigate the involvement of Rho-kinase and the actin filament network in Ang II- and pressure induced ERK1/2 activation, as well as in the contractile response induced by Ang II. Mesenteric resistance arteries (200 to 300 microm) were isolated, mounted in an arteriograph, and stimulated by pressure, Ang II, or both. Activation of ERK1/2 was then measured by an in-gel assay. In mesenteric resistance arteries maintained at 70 mm Hg, Ang II (0.1 micromol/L) induced contraction (29+/-1.4% of phenylephrine, 10 micromol/L-induced contraction) and significantly increased ERK1/2 activity. Selective inhibition of Rho-kinase by Y-27632 (10 micromol/L) and selective disruption of the actin filament network by cytochalasin B (10 micromol/L) both decreased the Ang II-induced contraction by 78+/-1.2% and 87+/ 1.9%, respectively, and significantly diminished ERK1/2 activity. In the absence of Ang II, increasing intraluminal pressure from 0 to 70 or 120 mm Hg increased ERK1/2 activity. ERK1/2 activity at 120 mm Hg was similar to that observed at 70 mm Hg in the presence of Ang II. Pressure-induced ERK1/2 activation was markedly attenuated by cytochalasin B but not by Y-27632. These results indicate that whereas pressure-induced ERK1/2 activation requires an intact actin filament network, but not Rho-kinase, the activation of ERK1/2 and the contraction induced by Ang II require both Rho-kinase and an intact actin filament network in isolated, intact mesenteric resistance arteries. PMID- 11498456 TI - Population of the vessel wall by leukocytes binding to P-selectin in a model of disturbed arterial flow. AB - We examined the hypothesis that disturbance of laminar flow promotes the attachment of leukocytes to the vessel wall in regions where the wall shear stress is otherwise too high. Isolated neutrophils, lymphocytes, or monocytes were perfused through chambers with backward-facing steps so that vortices occurred with well-defined reattachment of flow. Wall shear stresses downstream in reestablished flow equaled 0.07 Pa (low shear) or 0.3 Pa (high shear). In chambers coated with P-selectin, adherent leukocytes rolled. By use of a P selectin-Fc fragment chimera, adhesion was predominantly stationary, enabling definition of initial attachment sites. Neutrophils adhered in all regions of the low-shear chamber, with a local maximum around the reattachment point. However, in the high-shear chamber, adhesion was restricted to the recirculation zone and immediately downstream from the reattachment point. Rolling at high shear stress allowed a population of regions where initial attachment could not occur. At high shear, lymphocytes and monocytes also formed attachments restricted to the region of the reattachment point. The results imply that all types of leukocytes might bind to a capture receptor in high-shear vessels with discontinuities in the wall and might then spread to other regions. PMID- 11498457 TI - CD11b/CD18 mediates production of reactive oxygen species by mouse and human macrophages adherent to matrixes containing oxidized LDL. AB - Production of reactive oxygen species (ROS) and other proinflammatory substances by macrophages adherent to matrix proteins that contain or have been modified by oxidized LDL (oxLDL) may play an important role in atherogenesis. In vitro, human macrophages adhere to matrixes containing oxLDL via scavenger receptors and are signaled to produce ROS partly by interactions of the class B scavenger receptor (SR-B) CD36 with ligands on the matrix. In this report, we show that macrophages from mice genetically deficient in SR-A or CD36 adhered equally as well and produced equal amounts of ROS on interaction with matrix-associated oxLDL. In contrast, macrophages from mice genetically deficient in the CD18 chain of beta(2)-integrins produced insignificant amounts of ROS on interaction with oxLDL containing matrixes, even though they adhered to these matrixes as efficiently as did macrophages from wild-type mice. Antibodies against CD18, CD11b, or EDTA, the last of which chelates divalent cations required for integrin function, had no effect on adhesion of normal mouse or human macrophages to matrixes containing oxLDL but almost completely inhibited ROS production by macrophages adherent to this matrix. Thus, CD11b/CD18 plays an important role in regulating production of ROS by mouse and human macrophages adherent to matrixes containing oxLDL. It may play a hitherto-unsuspected role in regulating macrophage signaling pathways involved in inflammation and atherogenesis. PMID- 11498458 TI - Electron paramagnetic resonance investigation on modulatory effect of 17beta estradiol on membrane fluidity of erythrocytes in postmenopausal women. AB - Many studies have shown that estrogen may exert cardioprotective effects and reduce the risk of hypertension and coronary events. On the other hand, it has been proposed that cell membrane abnormalities play a role in the pathophysiology of hypertension, although it is not clear whether estrogen would influence membrane function in essential hypertension. The present study was performed to investigate the effects of 17beta-estradiol (E(2)) on membrane fluidity of erythrocytes in normotensive and hypertensive postmenopausal women. We determined the membrane fluidity of erythrocytes by means of an electron paramagnetic resonance and spin-labeling method. In an in vitro study, E(2) significantly decreased the order parameter for 5-nitroxide stearate and the peak height ratio for 16-nitroxide stearate obtained from electron paramagnetic resonance spectra of erythrocyte membranes in normotensive postmenopausal women. The finding indicates that E(2) might increase the membrane fluidity of erythrocytes. The effect of E(2) was significantly potentiated by the NO donor, S-nitroso-N acetylpenicillamine, and a cGMP analogue, 8-bromo-cGMP. In contrast, the change in the membrane fluidity evoked by E(2) was attenuated in the presence of the NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester, and asymmetric dimethyl-L arginine. In hypertensive postmenopausal women, the membrane fluidity of erythrocytes was significantly lower than that in normotensive postmenopausal women. The effect of E(2) on membrane fluidity was significantly more pronounced in the erythrocytes of hypertensive postmenopausal women than in the erythrocytes of normotensive postmenopausal women. The results of the present study showed that E(2) significantly increased the membrane fluidity and improved the microviscosity of erythrocyte membranes, partially mediated by an NO- and cGMP dependent pathway. Furthermore, the greater action of E(2) in hypertension might be consistent with the hypothesis that E(2) could have a beneficial effect in regulating rheological behavior of erythrocytes and could have a crucial role in the improvement of the microcirculation in hypertension. PMID- 11498459 TI - ACE genotype and endothelium-dependent vasodilation of conduit arteries and forearm microcirculation in humans. AB - The ACE gene is a candidate gene for cardiovascular disease. Endothelial dysfunction is considered an intermediate phenotype in the pathogenesis of hypertension and atherosclerosis. We evaluated the role of ACE gene polymorphism in endothelial function of young healthy humans. We assessed ACE genotype (deletion [D]/insertion [I] polymorphism) in 92 young healthy individuals. In 88 of them, endothelium-dependent (flow-mediated) vasodilation and endothelium independent (nitroglycerin-induced) vasodilation were measured in the common femoral artery and in the brachial (n=84) artery by echo Doppler technique. In 35 subjects, we also applied the forearm perfusion technique to quantify the responses of the forearm vascular bed to 3 increasing doses of 2 endothelium dependent vasodilators (acetylcholine and bradykinin) and 1 endothelium independent vasodilator (sodium nitroprusside). The D allele of the ACE gene was associated with a significant blunting (Delta approximately 26%) of endothelium dependent vasodilation in the femoral artery (P=0.02) but not in the brachial artery (P=0.55) or in the forearm microcirculation (P=0.70 to 0.80). Endothelium independent vasodilation was unaffected by the ACE genotype. In young healthy humans, the D allele of the ACE gene is associated with selective endothelial dysfunction of the femoral artery. It remains to be determined whether this association discloses a causal role in vascular, particularly peripheral artery, disease. PMID- 11498460 TI - Antioxidant supplements block the response of HDL to simvastatin-niacin therapy in patients with coronary artery disease and low HDL. AB - One strategy for treating coronary artery disease (CAD) patients with low HDL cholesterol (HDL-C) is to maximally increase the HDL-C to LDL-C ratio by combining lifestyle changes with niacin (N) plus a statin. Because HDL can prevent LDL oxidation, the low-HDL state also may benefit clinically from supplemental antioxidants. Lipoprotein changes over 12 months were studied in 153 CAD subjects with low HDL-C randomized to take simvastatin and niacin (S-N), antioxidants (vitamins E and C, beta-carotene, and selenium), S-N plus antioxidants (S-N+A), or placebo. Mean baseline plasma cholesterol, triglyceride, LDL-C, and HDL-C levels of the 153 subjects were 196, 207, 127, and 32 mg/dL, respectively. Without S-N, lipid changes were minor. The S-N and S-N+A groups had comparably significant reductions (P/=4.13 mmol/L and triglycerides <2.24 mmol/L) and in 90 normolipidemic subjects. Lp(a) levels were significantly elevated in patients compared with control subjects (0.35+/-0.4 and 0.24+/-0.31 mg/mL, respectively; median 0.13 and 0.11 mg/mL, respectively; P=0.039), although apo(a) isoform distribution did not differ. Patients displayed significantly higher plasma and urinary apo(a) fragment levels than did control subjects (respective values were as follows: 4.97+/-5.51 and 2.15+/-2.57 [median 2.85 and 1.17] microg/mL in plasma, P<0.0001; 75+/-86 and 40+/-57 [median 38 and 17] ng/mg urinary creatinine in urine, P<0.0001). The ratio of plasma apo(a) fragments to Lp(a) levels was also significantly higher in patients than in control subjects (1.93+/-1.5% and 1.75+/ 2.36%, respectively; P<0.0001). We conclude that increased plasma Lp(a) levels in polygenic hypercholesterolemia are associated with elevated circulating levels of apo(a) fragments but that this increase is not due to decreased renal clearance of apo(a) fragments. Furthermore, we identified a new pattern of apo(a) fragmentation characterized by the predominance of a fragment band whose size was related to that of the parent apo(a) isoform and that was superimposed on the series of fragments described previously by Mooser et al (J Clin Invest. 1996; 98:2414-2424). This new pattern was associated with small apo(a) isoforms and did not discriminate between hypercholesterolemic and normal subjects. However, this new apo(a) fragment pattern may constitute a novel marker for cardiovascular risk. PMID- 11498466 TI - Compensatory enlargement and stenosis develop in apoE(-/-) and apoE*3-Leiden transgenic mice. AB - Atherosclerotic mouse models develop little ischemic organ damage and no infarctions, despite the presence of large atherosclerotic lesions. Therefore, we hypothesize that luminal changes do not follow atherosclerotic lesion development. Because a phenomenon that may explain the discrepancy between luminal changes and lesion size is vascular remodeling, we measured parameters of vascular remodeling in the carotid arteries (CAs), thoracic aorta (TA), and abdominal aorta (AA) of apolipoprotein E (apoE)-deficient (apoE(-/-)) and apoE*3 Leiden mice, 2 well-known mouse models of atherosclerosis. Atherosclerotic lesions were classified (American Heart Association [AHA] types II through V), and plaque thickness, compensatory enlargement versus constrictive remodeling, lumen diameter, stenosis, and media thickness were measured relative to the nondiseased arterial wall. In CAs, plaque thickness increased during atherogenesis. CAs showed compensatory enlargement (apoE(-/-) 55%, apoE*3-Leiden 38%). Regression analysis revealed a positive correlation between plaque and lumen area (for apoE(-/-), R=0.95; for apoE*3-Leiden, R=0.90). Medial thinning and elastolysis were also observed. During atherogenesis, lumen diameter decreased (apoE(-/-) -69%, apoE*3-Leiden -40%), and stenosis >70% developed. TA and AA showed similar features, but neither developed a progressive decrease in lumen diameter or stenosis >70%. In CAs, TA, and AA of apoE(-/-) and apoE*3 Leiden mice, atherogenesis is associated with compensatory enlargement, medial thinning, and elastolysis. A progressive decrease in lumen diameter and stenoses >70% occur only in CAs. Vascular remodeling is more prominent in apoE(-/-) mice. PMID- 11498467 TI - Mice expressing only the mutant APOE3Leiden gene show impaired VLDL secretion. AB - Apolipoprotein E (apoE)-deficient mice develop hepatic steatosis and show impaired very low density lipoprotein (VLDL)-triglyceride (TG) secretion. These effects are normalized on the introduction of the human APOE3 gene. To assess whether this apoE effect is isoform specific, we studied hepatic lipid metabolism in mice expressing either APOE3 or the mutant APOE3Leiden on apoe-/- or apoe+/- backgrounds. The transgenes were expressed mainly in periportal hepatocytes, as revealed by in situ hybridization. Mice expressing APOE3Leiden, on the apoe-/- and apoe+/- backgrounds, had fatty livers, which were absent in APOE3/apoe-/- mice. APOE3Leiden/apoe-/- mice showed a strongly reduced VLDL-TG secretion compared with APOE3/apoe-/- mice (48+/-14 versus 82+/-10 micromol/kg per hour, respectively). The presence of a single mouse apoe allele increased VLDL-TG secretion in APOE3Leiden/apoe+/- mice (121+/-43 micromol/kg per hour) compared with APOE3Leiden/apoe-/- mice. These results show that APOE3Leiden does not prevent development of a fatty liver and does not normalize VLDL-TG secretion in mice with an apoE-deficient background. The presence of a single mouse apoe allele is sufficient to normalize the APOE3Leiden-associated reduction of VLDL-TG secretion but does not prevent steatosis. We conclude that apoE-mediated stimulation of VLDL secretion is isoform specific. PMID- 11498468 TI - Increased expression of heme oxygenase-1 and bilirubin accumulation in foam cells of rabbit atherosclerotic lesions. AB - Heme oxygenase-1 (HO-1) catalyzes the regiospecific oxidative degradation of heme to biliverdin IXalpha, iron, and carbon monoxide. Biliverdin IXalpha is subsequently reduced to bilirubin IXalpha by biliverdin reductase. HO-1 expression is induced under various disease conditions, including atherosclerosis, but it is unknown whether HO-1 catalyzes heme breakdown in the regions at risk. Using hypercholesterolemic rabbits fed a cholesterol-enriched diet, we attempted to demonstrate the involvement of HO-1 induction and bilirubin IXalpha production in atherosclerotic regions. Expression levels of HO-1 mRNA were elevated in the aortas of hypercholesterolemic rabbits. In situ hybridization and immunohistochemistry revealed that mRNA and protein of HO-1 are induced in endothelial cells and foam cells (lipid-filled macrophages) in atherosclerotic lesions. Furthermore, immunohistochemistry with the use of an anti-bilirubin-IXalpha monoclonal antibody, 24G7, demonstrated accumulation of bilirubin IXalpha in foam cells, indicating that heme is actually degraded in atherosclerotic lesions. Remarkably, bilirubin IXalpha, like HO-1 protein, is predominantly accumulated in the perinuclear regions of foam cells. These results provide the first in vivo evidence of the colocalization of HO-1 and bilirubin IXalpha in foam cells, suggesting a role of HO-1 induction in the modulation of macrophage activation in atherosclerosis. PMID- 11498469 TI - Plasminogen activator inhibitor type 1 is increased in the arterial wall of type II diabetic subjects. AB - Plasma plasminogen activator inhibitor type 1 (PAI-1) increases in diabetes, and this might contribute to decreased fibrinolysis and accelerated atherosclerosis. Increased PAI-1 levels in the vessel wall could decrease local fibrinolysis and elevate thrombus formation and the unfavorable evolution of atherosclerotic plaques. High glucose increases PAI-1 synthesis in arterial wall cells in culture, and aortic wall PAI-1 levels have been found to be elevated in diabetic animals. However, arterial wall PAI-1 levels have not been investigated in diabetic subjects. Therefore, the aim of this study was to determine the effect of diabetes on PAI-1 levels in the arterial wall. Blood samples and small tissue specimens from the mammary artery were obtained from 11 diabetic and 10 nondiabetic subjects who underwent coronary artery bypass graft surgery. PAI-1 antigen localization in the arterial wall was obtained by immunohistochemistry and was read by laser scanning confocal microscopy; plasma fibrinolytic activity was measured by lysis of fibrin plates; and PAI-1 activity was assessed by a chromogenic method. PAI-1-related immunofluorescence was increased in the arterial wall of diabetic patients, whereas plasma fibrinolysis was reduced. These data provide evidence that diabetes is associated with increased PAI-1 in the arterial wall. This might be an important factor for increased cardiovascular risk and unfavorable plaque evolution in diabetes. PMID- 11498470 TI - No relationship between compensatory arterial remodeling of focal stenotic atherosclerotic lesions and tortuosity of the arterial segment involved. PMID- 11498471 TI - Management of nodular thyroid disease. The challenge remains identifying which palpable nodules are malignant. PMID- 11498472 TI - The Campbell Collaboration. Does for public policy what cochrane does for health. PMID- 11498473 TI - Measuring the efficiency of health systems. The world health report sets the agenda, but there's still a long way to go. PMID- 11498474 TI - Fabricated or induced illness in children. Munchausen by proxy comes of age. PMID- 11498475 TI - Improving patients' safety by gathering information. Anonymous reporting has an important role. PMID- 11498476 TI - Indian doctors defend "unethical" anticancer drug trial. PMID- 11498477 TI - Second US institute investigates use of drug in asthma trial. PMID- 11498478 TI - UK civil rights groups question safety of stun guns. PMID- 11498479 TI - MMR vaccine row raises questions of clinical freedom. PMID- 11498480 TI - Researchers deny any attempt to mislead the public over JAMA article on arthritis drug. PMID- 11498481 TI - AIDS campaigners to take South Africa's health ministry to court. PMID- 11498482 TI - Asthma "crisis" for black Americans. PMID- 11498483 TI - New child health tsar promises shake-up of services. PMID- 11498484 TI - US tries to ban human cloning. PMID- 11498485 TI - Ecological study of effect of breast feeding on infant mortality in Latin America. AB - OBJECTIVE: To estimate the effect of exclusive breast feeding and partial breast feeding on infant mortality from diarrhoeal disease and acute respiratory infections in Latin America. DESIGN: Attributable fraction analysis of national data on infant mortality and breast feeding. SETTING: Latin America and the Caribbean. MAIN OUTCOME MEASURES: Mortality from diarrhoeal disease and acute respiratory infections and nationally representative breastfeeding rates. RESULTS: 55% of infant deaths from diarrhoeal disease and acute respiratory infections in Latin America are preventable by exclusive breast feeding among infants aged 0-3 months and partial breast feeding throughout the remainder of infancy. Among infants aged 0-3 months, 66% of deaths from these causes are preventable by exclusive breast feeding; among infants aged 4-11 months, 32% of such deaths are preventable by partial breast feeding. 13.9% of infant deaths from all causes are preventable by these breastfeeding patterns. The annual number of preventable deaths is about 52 000 for the region. CONCLUSIONS: Exclusive breast feeding of infants aged 0-3 months and partial breast feeding throughout the remainder of infancy could substantially reduce infant mortality in Latin America. Interventions to promote breast feeding should target younger infants. PMID- 11498486 TI - Comparative efficiency of national health systems: cross national econometric analysis. AB - OBJECTIVE: To improve the evidence base for health policy by devising a method to measure and monitor the performance of health systems. DESIGN: Estimation of the relation between levels of population health and the inputs used to produce health. SETTING: 191 countries. MAIN OUTCOME MEASURE: Health system efficiency (performance). RESULTS: Estimated efficiency varied from nearly fully efficient to nearly fully inefficient. Countries with a history of civil conflict or high prevalence of HIV and AIDS were less efficient. Performance increased with health expenditure per capita. CONCLUSIONS: Increasing the resources for health systems is critical to improving health in poor countries, but important gains can be made in most countries by using existing resources more efficiently. PMID- 11498487 TI - Influence of variation in birth weight within normal range and within sibships on IQ at age 7 years: cohort study. AB - OBJECTIVE: To examine the relation between birth weight and measured intelligence at age 7 years in children within the normal range of birth weight and in siblings. DESIGN: Cohort study of siblings of the same sex. SETTING: 12 cities in the United States. SUBJECTS: 3484 children of 1683 mothers in a birth cohort study during the years 1959 through 1966. The sample was restricted to children born at >/=37 weeks gestation and with birth weights of 1500-3999 g. MAIN OUTCOME MEASURE: Full scale IQ at age 7 years. RESULTS: Mean IQ increased monotonically with birth weight in both sexes across the range of birth weight in a linear regression analysis of one randomly selected sibling per family (n= 1683) with adjustment for maternal age, race, education, socioeconomic status, and birth order. Within same sex sibling pairs, differences in birth weight were directly associated with differences in IQ in boys (812 pairs, predicted IQ difference per 100 g change in birth weight =0.50, 95% confidence interval 0.28 to 0.71) but not girls (871 pairs, 0.10, -0.09 to 0.30). The effect in boys remained after differences in birth order, maternal smoking, and head circumference were adjusted for and in an analysis restricted to children with birth weight >/= 2500 g. CONCLUSION: The increase in childhood IQ with birth weight continues well into the normal birth weight range. For boys this relation holds within same sex sibships and therefore cannot be explained by confounding from family social environment. PMID- 11498488 TI - Simultaneous zinc and vitamin A supplementation in Bangladeshi children: randomised double blind controlled trial. AB - OBJECTIVE: To evaluate the effect of simultaneous zinc and vitamin A supplementation on diarrhoea and acute lower respiratory infections in children. STUDY DESIGN: Randomised double blind placebo controlled trial. SETTING: Urban slums of Dhaka, Bangladesh. PARTICIPANTS AND METHODS: 800 children aged 12-35 months were randomly assigned to one of four intervention groups: 20 mg zinc once daily for 14 days; 200 000 IU vitamin A, single dose on day 14; both zinc and vitamin A; placebo. The children were followed up once a week for six months, and morbidity information was collected. RESULTS: The incidence and prevalence of diarrhoea were lower in the zinc and vitamin A groups than in the placebo group. Zinc and vitamin A interaction had a rate ratio (95% confidence interval) of 0.79 (0.66 to 0.94) for the prevalence of persistent diarrhoea and 0.80 (0.67 to 0.95) for dysentery. Incidence (1.62; 1.16 to 2.25) and prevalence (2.07; 1.76 to 2.44) of acute lower respiratory infection were significantly higher in the zinc group than in the placebo group. The interaction term had rate ratios of 0.75 (0.46 to 1.20) for incidence and 0.58 (0.46 to 0.73) for prevalence of acute lower respiratory infection. CONCLUSIONS: Combined zinc and vitamin A synergistically reduced the prevalence of persistent diarrhoea and dysentery. Zinc was associated with a significant increase in acute lower respiratory infection, but this adverse effect was reduced by the interaction between zinc and vitamin A. PMID- 11498489 TI - Failure to report ethical approval in child health research: review of published papers. PMID- 11498490 TI - Do single handed practices offer poorer care? Cross sectional survey of processes and outcomes. AB - OBJECTIVES: To determine whether there are important differences in performance between group practices and singlehanded general practitioners and the extent to which any differences are explained by practice characteristics such as deprivation. DESIGN: Cross sectional survey. SETTING: 206 singlehanded practices and 606 partnerships in Trent region, United Kingdom. METHOD: Comparison of process and outcome measures derived from routinely collected data on hospital admissions and target payments for singlehanded practices and partnerships. Multivariate analysis was used to adjust for the confounding effects of general practice characteristics-deprivation (Townsend score), percentage of Asian residents, percentage of black residents, proportion of men over 75 years, proportion of women over 75 years, rurality, presence of a female general practitioner, and vocational training status. RESULTS: Differences in achievement of immunisation and cytology targets apparent on univariate analysis were not seen after adjustment for other general practice characteristics. Similarly, significant differences (>15%; P<0.01) for three types of hospital admission seen on univariate analysis were not present after adjustment for other practice characteristics. CONCLUSIONS: This study provides no evidence that singlehanded general practitioners are underperforming clinically. Our results offer insight into the structural difference between the two types of practice and underline the importance of the effect of other practice characteristics on process and outcome measures. PMID- 11498491 TI - Use of cumulative mortality data in patients with acute myocardial infarction for early detection of variation in clinical practice: observational study. AB - OBJECTIVES: Use of cumulative mortality adjusted for case mix in patients with acute myocardial infarction for early detection of variation in clinical practice. DESIGN: Observational study. SETTING: 20 hospitals across the former Yorkshire region. PARTICIPANTS: All 2153 consecutive patients with confirmed acute myocardial infarction identified during three months. MAIN OUTCOME MEASURES: Variable life-adjusted displays showing cumulative differences between observed and expected mortality of patients; expected mortality calculated from risk model based on admission characteristics of age, heart rate, and systolic blood pressure. RESULTS: The performance of two individual hospitals over three months was examined as an example. One, the smallest district hospital in the region, had a series of 30 consecutive patients but had five more deaths than predicted. The variable life-adjusted display showed minimal variation from that predicted for the first 15 patients followed by a run of unexpectedly high mortality. The second example was the main tertiary referral centre for the region, which admitted 188 consecutive patients. The display showed a period of apparently poor performance followed by substantial improvement, where the plot rose steadily from a cumulative net lives saved of -4 to 7. These variations in patient outcome are unlikely to have been revealed during conventional audit practice. CONCLUSIONS: Variable life-adjusted display has been integrated into surgical care as a graphical display of risk-adjusted survival for individual surgeons or centres. In combination with a simple risk model, it may have a role in monitoring performance and outcome in patients with acute myocardial infarction. PMID- 11498493 TI - Recent advances: Sports medicine. PMID- 11498494 TI - Lesson of the week: Deterioration of symptoms after start of thyroid hormone replacement. PMID- 11498496 TI - A new system for grading recommendations in evidence based guidelines. PMID- 11498503 TI - Protein kinase A-independent responses to beta-adrenoceptor agonists. PMID- 11498504 TI - Evidence that the anti-spasmogenic effect of the beta-adrenoceptor agonist, isoprenaline, on guinea-pig trachealis is not mediated by cyclic AMP-dependent protein kinase. AB - 1. The spasmolytic and anti-spasmogenic activity of beta-adrenoceptor agonists on airways smooth muscle is thought to involve activation of the cyclic AMP/cyclic AMP-dependent protein kinase (PKA) cascade. Here we have tested the hypothesis that PKA mediates the anti-spasmogenic activity of isoprenaline and other cyclic AMP-elevating agents in guinea-pig isolated trachea by utilizing a number of cell permeant cyclic AMP analogues that act as competitive 'antagonists' of PKA. 2. Anion-exchange chromatography of guinea-pig tracheae resolved two peaks of PKA activity that corresponded to the type I ( approximately 5%) and type II ( approximately 93%) isoenzymes. 3. Pre-treatment of tracheae with zardaverine (30 microM), vasoactive intestinal peptide (VIP) (1 microM) and the non-selective activator of PKA, Sp-8-CPT-cAMPS (10 microM), produced a non-parallel rightwards shift in the concentration-response curves that described acetylcholine (ACh) induced tension generation. The type II-selective PKA inhibitor, Rp-8-CPT-cAMPS (300 microM), abolished this effect. 4. Pre-treatment of tracheae with Sp-8-Br PET-cGMPS (30 microM) produced a non-parallel rightwards shift of the concentration-response curves that described ACh-induced tension generation. The selective cyclic GMP-dependent protein kinase (PKG) inhibitor, Rp-8-pCPT-cGMPS (300 microM), abolished this effect. 5. Pre-treatment of tracheae with isoprenaline (1 microM) produced a 10 fold shift to the right of the ACh concentration-response curve by a mechanism that was unaffected by Rp-8-Br-cAMPS (300 microM, selective inhibitor of type I PKA), Rp-8-CPT-cAMPS (300 microM) and Rp-8-pCPT-cGMPS (300 microM). 6. We conclude that the anti-spasmogenic activity of Sp-8-CPT-cAMPS, zardaverine and VIP in guinea-pig trachea is attributable to activation of the cyclic AMP/PKA cascade whereas isoprenaline suppresses ACh induced contractions by a mechanism(s) that is independent of PKA and PKG. PMID- 11498505 TI - In vivo evidence that protease-activated receptors 1 and 2 modulate gastrointestinal transit in the mouse. AB - 1. Protease-activated receptors (PARs) 1 and 2 modulate the gastric and intestinal smooth muscle motility in vitro. In the present study, we examined if activation of PAR-2 and PAR-1 could alter gastrointestinal transit in mice. 2. Intraperitoneal administration of the PAR-2-activating peptide SLIGRL-NH(2), but not the inactive control LSIGRL-NH(2), at 1 - 5 micromol kg(-1), in combination with the aminopeptidase inhibitor amastatin at 2.5 micromol kg(-1), facilitated gastrointestinal transit in a dose-dependent manner. The human PAR-1-derived peptide SFLLR-NH(2) and the specific PAR-1 agonist TFLLR-NH(2), but not the inactive control FSLLR-NH(2), at 2.5 - 10 micromol kg(-1), in combination with amastatin, also promoted gastrointestinal transit. 3. The Ca2+-activated, small conductance K+ channel inhibitor apamin at 0.01 micromol kg(-1) significantly potentiated the actions of SLIGRL-NH(2) and TFLLR-NH(2) at subeffective doses. 4. The increased gastrointestinal transit exerted by either SLIGRL-NH(2) at 5 micromol kg(-1) or TFLLR-NH(2) at 10 micromol kg(-1) was completely abolished by the L-type Ca2+ channel inhibitor verapamil at 61.6 micromol kg(-1). In contrast, the tyrosine kinase inhibitor genistein at 18.5 micromol kg(-1) failed to modify the effects of the agonists for PAR-2 or PAR-1. 5. These findings demonstrate that PAR-1 and PAR-2 modulate gastrointestinal transit in mice in vivo. Our data also suggest that the PAR-1-and PAR-2-mediated effects are modulated by apamin sensitive K+ channels and are dependent on activation of L-type Ca2+ channels, but independent of tyrosine kinase. Our study thus provides novel evidence for the physiological and/or pathophysiological roles of PARs 1 and 2 in the digestive systems, most probably during inflammation. PMID- 11498506 TI - Inactivation of platelet-derived growth factor-BB following modification by ADP ribosyltransferase. AB - 1. Arginine-specific ADP-ribosyltransferase (ART1) is expressed on the surface of a number of cell types, and catalyses the transfer of ADP-ribose from NAD(+) to target proteins. We investigated whether extracellular proteins such as growth factors may serve as substrates for this enzyme, with subsequent alteration in their biological activity. Experiments were performed with rat skeletal muscle membranes and V79 Chinese hamster lung fibroblasts with doxycycline-inducible expression of human ART. 2. From a panel of growth factors, platelet-derived growth factor-BB (PDGF-BB) was found to be the best substrate for ART1, whereas the structural homologue PDGF-AA was not a substrate. Under conditions of maximum labelling 5 mol ADP-ribose was incorporated per mol of PDGF-BB. 3. Purified (ADP ribosyl)-PDGF-BB did not stimulate a mitogenic or chemotactic response in human pulmonary smooth muscle cells, and showed a reduced capacity to bind to PDGF receptors in competition binding experiments, when compared to unmodified PDGF BB. 4. PDGF-dependent [(3)H-methyl]-thymidine incorporation was measured in the ART1-transfected fibroblast cell line at physiological concentrations of PDGF-BB, and without addition of extracellular NAD(+). Fibroblasts expressing human ART1 at the cell surface showed reduced mitogenic responses to PDGF-BB, but not to PDGF-AA. This loss of mitogenic response in cells expressing ART1 activity was reversed by the addition of agmatine (an ART1 substrate). 5. In conclusion, we propose that PDGF-BB-dependent signalling may be regulated by post-translational modification of the growth factor by ART1 at the cell surface. This has been demonstrated in membranes of rat skeletal muscle, and the reaction confirmed in ART1-transfected fibroblasts. PMID- 11498507 TI - The cholecystokinin analogues JMV-180 and CCK-8 stimulate phospholipase C through the same binding site of CCK(A) receptor in rat pancreatic acini. AB - 1. This study was designed to address the controversy related to the involvement of phospholipase C in the signalling pathway linked to CCK(A) receptor stimulation by the cholecystokinin analogue JMV-180, a full agonist for amylase release, in rat pancreatic acini. 2. JMV-180 was shown to stimulate phospholipase C by measuring the incorporation of [(32)P]-orthophosphoric acid ([(32)P]-Pi) into phosphatidic acid (PtdOH) and phosphatidylinositol (PtdIns). Both responses elicited by JMV-180 were time and concentration dependent. Maximal effects elicited by JMV-180 were 39.08+/-0.72 and 8.02+/-0.40% for the labelling of [(32)P]-PtdIns and [(32)P]-PtdOH, respectively, as compared to the maximal effects of CCK-8, a full agonist of the CCK(A) receptor. 3. [(32)P]-Pi incorporation into PtdOH and PtdIns was sensitive to lithium, demonstrating that both responses are a consequence of phospholipase C activation. However, since lithium blocks the phosphoinositide cycle by an uncompetitive mechanism, its effect was only apparent at high concentrations of CCK-8 (>10 pM), which elicited stimuli above 20 and 60% of the maximal [(32)P]-PtdOH and [(32)P]-PtdIns labelling, respectively. 4. JMV-180 inhibited the incorporation of [(32)P]-Pi into PtdOH and PtdIns as stimulated by CCK-8, down to its own maximal effect. The estimated IC(50) values for the inhibition curves were not significantly different from those calculated assuming the same single binding site for both agonists. These results indicated that the well established role of JMV-180 as a partial agonist for CCK(A) receptor-linked signalling responses, also applies for the stimulation of phospholipase C. 5. The comparison of CCK-8 and JMV-180 dose response curves of amylase release to those of PtdIns and PtdOH labelling with [(32)P]-Pi showed the existence of an amplification mechanism between phospholipase C and amylase release for both agonists. 6. In conclusion, we show that JMV-180, as well as CCK-8, stimulate phospholipase C upon interaction with the same binding site at the CCK(A) receptor in rat pancreatic acini. PMID- 11498508 TI - Role of nitric oxide and superoxide in allergen-induced airway hyperreactivity after the late asthmatic reaction in guinea-pigs. AB - 1. In the present study, the roles of nitric oxide (NO) and superoxide anions (O2(-)) in allergen-induced airway hyperreactivity (AHR) after the late asthmatic reaction (LAR) were investigated ex vivo, by examining the effects of the NO synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) and superoxide dismutase (SOD) on the responsiveness to methacholine of isolated perfused guinea pig tracheae from unchallenged (control) animals and from animals 24 h after ovalbumin challenge. 2. At 24 h after allergen challenge, the animals developed AHR in vivo, as indicated by a mean 2.63 +/- 0.54 fold (P < 0.05) increase in sensitivity to histamine inhalation. 3. Compared to unchallenged controls, tracheal preparations from the ovalbumin-challenged guinea-pigs displayed a significant 1.8 fold (P < 0.01) increase in the maximal response (E(max)) to methacholine, both after intraluminal (IL) and extraluminal (EL) administration of the agonist. No changes were observed in the sensitivity (pEC(50)) to the agonist. Consequently, the DeltapEC(50) (EL-IL), as a measure of epithelial integrity, was unchanged. 4. In the presence of L-NAME (100 microM, IL), tracheae from control guinea-pigs showed a 1.6 fold (P < 0.05) increase in the E(max) of IL methacholine. By contrast, the E(max) of IL methacholine was significantly decreased in the presence of 100 u ml(-1) EL SOD (54% of control, P < 0.01). 5. Remarkably, the increased responsiveness to IL methacholine at 24 h after allergen challenge was reversed by L-NAME to control (P < 0.01), and a similar effect was observed with SOD (P < 0.01). 6. The results indicate that both NO and O2(-) are involved in the tracheal hyperreactivity to methacholine after the LAR, possibly by promoting airway smooth muscle contraction through the formation of peroxynitrite. PMID- 11498509 TI - Identification of the dopamine autoreceptor in the guinea-pig retina as D(2) receptor using novel subtype-selective antagonists. AB - 1. Dopamine release in the retina is subject to modulation via autoreceptors, which belong to the D(2) receptor family (encompassing the D(2), D(3) and D(4) receptors). The aim of the present study was to determine the receptor subtype (D(2) vs D(3)) involved in the inhibition of dopamine release in guinea-pig retinal discs, using established (haloperidol, (S)-nafadotride) and novel dopamine receptor antagonists (ST-148, ST-198). 2. hD(2L) and hD(3) receptors were expressed in CHO cells and the pK(i) values determined in binding studies with [(125)I]-iodosulpride were: haloperidol 9.22 vs 8.54; ST-148 7.85 vs 6.60; (S)-nafadotride 8.52 vs 9.51; ST-198 6.14 vs 7.92. 3. The electrically evoked tritium overflow from retinal discs preincubated with [(3)H]-noradrenaline (which represents quasi-physiological dopamine release) was inhibited by the dopamine receptor agonists B-HT 920 (talipexole) and quinpirole (maximally by 82 and 71%; pEC(50) 5.80 and 5.83). The concentration-response curves of these agonists were shifted to the right by haloperidol (apparent pA(2) 8.69 and 8.23) and ST-148 (7.52 and 7.66). (S)-Nafadotride 0.01 microM and ST-198 0.32 microM did not affect the concentration-response curve of B-HT 920. 4. The dopamine autoreceptor in the guinea-pig retina can be classified as a D(2) receptor. ST-148 and ST-198 show an improved selectivity for D(2) and D(3) receptors when compared to haloperidol and (S)-nafadotride, respectively. PMID- 11498510 TI - Mutational analysis of the human vasoactive intestinal peptide receptor subtype VPAC(2): role of basic residues in the second transmembrane helix. AB - 1. We investigated the role of two conserved basic residues in the second transmembrane helix arginine 172 (R172) and lysine 179 (K179) of the VPAC(2) receptor. 2. Vasoactive intestinal polypeptide (VIP) activated VPAC(2) receptors with an EC(50) value of 7 nM, as compared to 150, 190 and 4000 nM at R172L, R172Q and K179Q-VPAC(2) receptors, respectively. It was inactive at K179I mutated VPAC(2) receptors. These results suggested that both basic residues were probably implicated in receptor recognition and activation. 3. The VPAC(2)-selective VIP analogue, [hexanoyl-His(1)]-VIP (C(6)-VIP), had a higher affinity and efficacy as compared to VIP at the mutated receptors. 4. VIP, Asn(3)-VIP and Gln(3)-VIP activated adenylate cyclase through R172Q receptors with EC(50) values of 190, 2 and 2 nM, respectively, and through R172L receptors with EC(50) values of 150, 12 and 8 nM, respectively. Asn(3)-VIP and Gln(3)-VIP behaved as partial agonists at the wild type receptor, with E(max) values (in per cent of VIP) of 75 and 52%, respectively. In contrast, they were more efficient than VIP (E(max) values of 150 and 150% at the R172Q VPAC(2) receptors, and of 400 and 360% at the R172L receptors, respectively). These results suggested that the receptor's R172 and the ligand's aspartate 3 are brought in close proximity in the active ligand receptor complex. 5. The K179I and K179Q mutated receptors had a lower affinity than the wild-type receptors for all the agonists tested in this work: we were unable to identify the VIP amino acid(s) that interact with K179. PMID- 11498511 TI - Inhibition of mechanical activation of guinea-pig airway afferent neurons by amiloride analogues. AB - 1. The aim of this study was to investigate a role for Epithelial Sodium Channels (ENaCs) in the mechanical activation of low-threshold vagal afferent nerve terminals in the guinea-pig trachea/bronchus. 2. Using extracellular single-unit recording techniques, we found that the ENaC blocker amiloride, and its analogues dimethylamiloride and benzamil caused a reduction in the mechanical activation of guinea-pig airway afferent fibres. 3. Amiloride and it analogues also reduced the sensitivity of afferent fibres to electrical stimulation such that greater stimulation voltages were required to induce action potentials from their peripheral terminals within the trachea/bronchus. 4. The relative potencies of these compounds for inhibiting electrical excitability of afferent nerves were similar to that observed for inhibition of mechanical stimulation (dimethylamiloride approximately benzamil > amiloride). This rank order of potency is incompatible with the known rank order of potency for blockade of ENaCs (benzamil > amiloride >> dimethylamiloride). 5. As voltage-gated sodium channels play an important role in determining the electrical excitability of neurons, we used whole-cell patch recordings of nodose neuron cell bodies to investigate the possibility that amiloride analogues caused blockade of these channels. At the concentration required to inhibit mechanical activation of vagal nodose afferent fibres (100 microM), benzamil caused significant inhibition of voltage-gated sodium currents in neuronal cell bodies acutely isolated from guinea-pig nodose ganglia. 6. Combined, our findings suggest that amiloride and its analogues did not selectively block mechanotransduction in airway afferent neurons, but rather they reduced neuronal excitability, possibly by inhibiting voltage-gated sodium currents. PMID- 11498512 TI - Effects of homologues and analogues of palmitoylethanolamide upon the inactivation of the endocannabinoid anandamide. AB - 1. The ability of a series of homologues and analogues of palmitoylethanolamide to inhibit the uptake and fatty acid amidohydrolase (FAAH)-catalysed hydrolysis of [(3)H]-anandamide ([(3)H]-AEA) has been investigated. 2. Palmitoylethanolamide and homologues with chain lengths from 12 - 18 carbon atoms inhibited rat brain [(3)H]-AEA metabolism with pI(50) values of approximately 5. Homologues with chain lengths < or = eight carbon atoms gave < 20% inhibition at 100 microM. 3. R palmitoyl-(2-methyl)ethanolamide, palmitoylisopropylamide and oleoylethanolamide inhibited [(3)H]-AEA metabolism with pI(50) values of 5.39 (competitive inhibition), 4.89 (mixed type inhibition) and 5.33 (mixed type inhibition), respectively. 4. With the exception of oleoylethanolamide, the compounds did not produce dramatic inhibition of [(3)H]-WIN 55,212-2 binding to human CB(2) receptors expressed on CHO cells. Palmitoylethanolamide, palmitoylisopropylamide and R-palmitoyl-(2-methyl)ethanolamide had modest effects upon [(3)H]-CP 55,940 binding to human CB(1) receptors expressed on CHO cells. 5. Most of the compounds had little effect upon the uptake of [(3)H]-AEA into C6 and/or RBL-2H3 cells. However, palmitoylcyclohexamide (100 microM) and palmitoylisopropylamide (30 and 100 microM) produced more inhibition of [(3)H]-AEA uptake than expected to result from inhibition of [(3)H]-AEA metabolism alone. 6. In intact C6 cells, palmitoylisopropylamide and oleoylethanolamide inhibited formation of [(3)H] ethanolamine from [(3)H]-AEA to a similar extent as AM404, whereas palmitoylethanolamide, palmitoylcyclohexamide and R-palmitoyl-(2 methyl)ethanolamide were less effective. 7. These data provide useful information upon the ability of palmitoylethanolamide analogues to act as 'entourage' compounds. Palmitoylisopropylamide may prove useful as a template for design of compounds that reduce the cellular accumulation and metabolism of AEA without affecting either CB(1) or CB(2) receptors. PMID- 11498513 TI - Mechanism of trypsin-induced contraction in the rat myometrium: the possible involvement of a novel member of protease-activated receptor. AB - 1. The mechanism of trypsin-induced contraction in the rat myometrium was investigated using front-surface fluorimetry on fura-PE3-loaded strips. The expression of protease-activated receptors (PARs) in the rat myometrium was determined by reverse transcription polymerase chain reaction (RT - PCR). 2. In non-pregnant rats, 10 microM trypsin developed a force of up to 30.5 +/- 5.1% of that obtained during the 40 mM K(+)-depolarization-induced contraction. In pregnant rats, the maximal level of the cytosolic Ca(2+) concentration and tension obtained with 3 microM trypsin was 143.2 +/- 6.0% and 63.2 +/- 7.9%, respectively. The depletion of the extracellular Ca2+ abolished the trypsin induced contraction. 3. Trypsin-induced contraction was abolished by the pre treatment of a serine protease inhibitor. PAR1-activating peptide (PAR1-AP) caused a potent contraction of the myometrium, while neither PAR2-AP nor PAR4-AP induced any contraction. 4. RT - PCR analysis detected the expression of PAR1 mRNA. However, neither PAR2 nor PAR4 mRNA was detected in the rat myometrium. 5. Once the strips were stimulated with thrombin, the subsequent application of thrombin failed to induce any contraction, while trypsin induced a contraction similar to that observed without the pre-stimulation with thrombin. Once the strip was stimulated with trypsin, neither trypsin nor thrombin induced any contraction. The response to PAR1-AP remained after the pre-stimulation with thrombin and trypsin. 6. In conclusion, PAR1 was the only known receptor for trypsin expressed in the rat myometrium, but it was suggested to be cleaved and inactivated by trypsin. Trypsin was thus suggested to contract the rat myometrium via a novel type of PAR, which might be upregulated during pregnancy. PMID- 11498514 TI - Differential effects of chronic drug treatment on alpha3* and alpha7 nicotinic receptor binding sites, in hippocampal neurones and SH-SY5Y cells. AB - 1. The aim of this study was to compare the effects of chronic treatment (for 4 or 7 days) with nicotinic drugs and 20 mM KCl on numbers of surface alpha7 nicotinic AChR, identified by [(125)I]-alpha bungarotoxin (alpha-Bgt) binding, in primary hippocampal cultures and SH-SY5Y cells. Numbers of alpha3* nicotinic AChR were also examined in SH-SY5Y cells, using [(3)H]-epibatidine, which is predicted to label the total cellular population of predominantly alpha3beta2* nicotinic AChR under the conditions used. 2. All the nicotinic agonists examined, the antagonists d-tubocurarine and methyllycaconitine, and KCl, upregulated [(125)I] alpha Bgt binding sites by 20 - 60% in hippocampal neurones and, where examined, SH-SY5Y cells. 3. Upregulation of [(125)I]-alpha-Bgt binding sites by KCl was prevented by co-incubation with the L-type Ca2+ channel blocker verapamil or the Ca2+-calmodulin dependent kinase II (CaM-kinase II) inhibitor KN-62. Upregulation of [(125)I]-alpha-Bgt binding sites by nicotine or 3,[(4-dimethylamino) cinnamylidene] anabaseine maleate (DMAC) was insensitive to these agents. 4. [(3)H]-Epibatidine binding sites in SH-SY5Y cells were not affected by KCl but were upregulated in a verapamil-insensitive manner by nicotine and DMAC. KN-62 itself provoked a 2 fold increase in [(3)H]-epibatidine binding. The inactive analogue KN-04 had no effect, suggesting that CaM-kinase II plays a role in regulating numbers of alpha3* nicotinic AChR. 5. These data indicate that numbers of alpha3* and alpha7 nicotinic AChR are modulated differently. Nicotinic agonists and KCl upregulate alpha7 nicotinic AChR through distinct cellular mechanisms, the latter involving L-type Ca2+ channels and CaM-kinase II. In contrast, alpha3* nicotinic AChR are not upregulated by KCl. This difference may reflect the distinct physiological roles proposed for alpha7 nicotinic AChR. PMID- 11498515 TI - Proinflammatory characteristics of a nonpeptide bradykinin mimic, FR190997, in vivo. AB - 1. Proinflammatory potency of the nonpeptide bradykinin (BK) B(2) receptor agonist FR190997 (8-[2,6-dichloro-3-[N-[(E)-4-(N-methylcarbamoyl)cinnamidoacetyl] N-methylamino]benzyloxy]-2-methyl-4-(2-pyridylmethoxy)quinoline) was investigated. 2. Intradermal injection of FR190997 (0.03 - 3 nmol site(-1)) into dorsal skin of rats increased vascular permeability in a dose-dependent manner. The effect was less than that of BK, but it was long-acting and was inhibited by treatment with FR173657 (3 mg kg(-1), p.o.). Captopril (10 mg kg(-1), i.p.) did not enhance the plasma extravasation by FR190997 (0.3 nmol site(-1)) in the presence of soybean trypsin inhibitor (SBTI, 30 microg site(-1)). 3. Subcutaneous injection of FR190997 (3 nmol site(-1)) into the hindpaw of mice markedly induced paw swelling. The oedema lasted up to 3 h after the injection. Administration of indomethacin or NS-398 (10 mg kg(-1), i.p.) significantly reduced it at 3 h after the injection. 4. Simultaneous i.p. injection of prostaglandin (PG) E(2) (1 nmol site(-1)) or beraprost sodium (0.5 nmol site(-1)) with FR190997 (5 nmol site(-1)) greatly enhanced frequency of writhing reactions in mice. 5. FR190997 (0.3 - 30 nmol kg(-1), i.v.) showed less increase in airway opening pressure (Pao) in the guinea-pig after i.v. injection. Furthermore, FR190997 (0.03 - 30 nmol) resulted in a very weak contraction of tracheal ring strips and lung parenchymal sections in vitro. 6. In mice sponge implants, topical application of FR190997 increased angiogenesis and granulation with enhanced expressions of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) mRNAs. 7. These results indicate that FR190997 has proinflammatory long-lasting characteristics and it might be 'a stable tool' for studying the role of BK B(2) receptor in vivo. PMID- 11498516 TI - Ca(2+)-sensitizing effect is involved in the positive inotropic effect of troglitazone. AB - 1. Troglitazone, an insulin sensitizing agent, has a direct positive inotropic effect. However, the mechanism of this effect remains unclear. Thus, we examined the inotropic effect of troglitazone while focusing on intracellular Ca2+ handling. 2. Troglitazone significantly increased peak isovolumic left ventricular pressure (LVP(max)), peak rate of rise of LVP (dP/dt(max)), peak rate of fall of LVP (dP/dt(min)) in isolated rat hearts perfused at a constant coronary flow and heart rate. This inotropic effect of troglitazone was not inhibited by pretreatment with carbachol (muscarine receptor agonist), H89 (protein kinase A inhibitor), U73122 (phospholipase C inhibitor), H7 (protein kinase C inhibitor), verapamil (L-type Ca2+ channel antagonist), thapsigargin (Ca(2+)-adenosine triphosphatase inhibitor) or ryanodine (ryanodine receptor opener). 3. Radioimmunoassay showed that the cyclic adenosine monophosphate concentration in the left ventricle was not increased by troglitazone. 4. Whole cell patch clamp analysis revealed that troglitazone had no effect on inward Ca2+ currents in cardiomyocytes. 5. In fura-2 loaded perfused rat hearts, troglitazone exerted its positive inotropic effect without increasing Ca2+ concentration. 6. These results suggest that neither the inward Ca2+ currents nor Ca2+ handling in the sarcoplasmic reticulum was involved in the inotropic effect of troglitazone. Furthermore, troglitazone exerted its positive inotropic effect without affecting the intracellular concentration of Ca2+. 7. In conclusion, the positive inotropic effect of troglitazone is mediated by a sensitization of Ca2+. PMID- 11498517 TI - Vasorelaxant effects of a nitric oxide-releasing aspirin derivative in normotensive and hypertensive rats. AB - 1. Nonsteroidal anti-inflammatory drugs have been reported to exacerbate hypertension and to interfere with the effectiveness of some anti-hypertensive therapies. In this study, we tested the effects of a gastric-sparing, nitric oxide-releasing derivative of aspirin (NCX-4016) on hypertension in rats. 2. Hypertension was induced by administering L-NAME in the drinking water (400 mg l( 1)). Groups of rats were treated daily with aspirin, NCX-4016 or vehicle. 3. NCX 4016 significantly reduced blood pressure relative to the aspirin-treated group over the 2-week period of treatment. Aspirin and, to a lesser extent, NCX-4016 suppressed whole blood thromboxane synthesis. 4. In anaesthetized rats, acute intravenous administration of NCX-4016 caused a significant fall in mean arterial pressure in hypertensive rats, but was devoid of such effects in normotensive controls. 5. In vitro, NCX-4016 relaxed phenylephrine-pre-contracted aortic rings obtained from both normotensive and hypertensive rats, and significantly reduced their responsiveness to the contractile effects of phenylephrine. 6. These results suggest that NCX-4016 reduces blood pressure in hypertensive rats, not simply through the direct vasodilatory actions of the nitric oxide released by this compound, but also through possible interference with the effects of endogenous pressor agents. These properties, added to its anti-thrombotic effects, suggest that NCX-4016 may be a safer alternative to aspirin for use by hypertensive patients. PMID- 11498518 TI - Production of leukotrienes in a model of focal cerebral ischaemia in the rat. AB - 1. The aim of this work was to evaluate the role of leukotrienes in brain damage in vivo in a model of focal cerebral ischaemia in the rat, obtained by permanent occlusion of middle cerebral artery. 2. A significant (P < 0.01) elevation of LTC(4), LTD(4) and LTE(4) (cysteinyl-leukotrienes) levels occurred 4 h after ischaemia induction in the ipsilateral cortices of ischaemic compared to sham operated animals (3998 +/- 475 and 897 +/- 170 fmol g(-1) tissue, respectively, P < 0.01). 3. The NMDA receptor antagonist MK-801 and the adenosine A(2A) receptor antagonist SCH 58261 were administered in vivo at doses known to reduce infarct size and compared with the leukotriene biosynthesis inhibitor MK-886. 4. MK-886 (0.3 and 2 mg kg(-1) i.v.) and MK-801 (3 mg kg(-1) i.p.) decreased cysteinyl leukotriene levels (-78%, P < 0.05; -100%, P < 0.01; -92%, P < 0.01, respectively) 4 h after permanent occlusion of the middle cerebral artery, whereas SCH 58261 (0.01 mg kg(-1) i.v.) had no significant effects. 5. MK-886 (2 mg kg(-1) i.v.) was also able to significantly reduce the cortical infarct size by 30% (P < 0.05). 6. We conclude that cysteinyl-leukotriene formation is associated with NMDA receptor activation, and that it represents a neurotoxic event, the inhibition of which is able to reduce brain infarct area in a focal ischaemic event. PMID- 11498519 TI - Nebivolol, bucindolol, metoprolol and carvedilol are devoid of intrinsic sympathomimetic activity in human myocardium. AB - 1. The present study investigated whether or not there may be differences in the direct cardiac actions of the novel, highly beta(1)-selective adrenoceptor antagonist nebivolol (NEB) in comparison to metoprolol (MET), bisoprolol (BIS), carvedilol (CAR) and bucindolol (BUC) in human myocardium (n=9). 2. The rank order of beta(1)-selectivity as judged by competition experiments to (3)H-CGP 12.1777 in the presence of CGP 207.12 A (300 nmol l(-1), K(i)beta(2)) or ICI 118.551 (50 nmol l(-1), K(i)beta(1)) were NEB(K(i)beta(2)/K(i)beta(1): 40.7) > BIS(15.6) > MET(4.23) > CAR(0.73) > BUC(0.49). 3. The rank order of the negative inotropic potency of the beta-adrenoceptor antagonists measured in left ventricular trabeculae (dilated cardiomyopathy, DCM) as judged by the concentration needed to induce a 50% decrease in isoprenaline (1 micromol l(-1)) stimulated force (IC(50)) was: MET (0.6 micromol l(-1)) > CAR (4.1 micromol l( 1)) > NEB (7.0 micromol l(-1)). 4. NEB, BUC, MET and CAR did not not exert an intrinsic sympathomimetic activity (ISA) as determined by measurements of force development in forskolin (0.3 micromol l(-1)) pre-treated left ventricular trabeculae, nor by measuring adenylate cyclase activity in forskolin (0.3 micromol l(-1))-stimulated assays (crude membranes). This also holds true for radioligand binding assays with or without guanine nucleotide guanyl-5'-yl imidodiphosphate (Gpp(NH)p). 5. Although all studied beta-adrenoceptor antagonists lack intrinsic sympathomimetic activity (ISA), they differ in the beta(1)-selectivity as well as in their direct negative inotropic action. These differences as well as the mode of extracardiac action may have an impact on outcome of patients treated with beta-adrenoceptor antagonists. PMID- 11498520 TI - Bradykinin increases Na(+)-K(+) pump activity in cultured guinea-pig tracheal smooth muscle cells. AB - 1. The effect of bradykinin on the Na+-K+ pump of airway smooth muscle was investigated by measuring ouabain-sensitive (86)Rb(+) uptake in cultured guinea pig tracheal smooth muscle cells. 2. Bradykinin induced a concentration-dependent increase in ouabain-sensitive (86)Rb(+) uptake, with an EC(50) of 3 nM (pD(2) = 8.50+/-0.10). Stimulation was not affected by indomethacin (1 microM) suggesting that it is not mediated by cycloxygenase products of arachidonic acid. 3. The B(1) receptor agonists Lys-des-Arg(9)-bradykinin and des-Arg(9)-bradykinin had no effect on ouabain-sensitive (86)Rb(+) uptake. In contrast, the B(1) and B(2) receptor agonist Lys-bradykinin induced a concentration-dependent increase in ouabain-sensitive (86)Rb(+) uptake with an EC(50) of 6 nM (pD(2) = 8.21 +/- 0.20). 4. The B(1) receptor antagonist des-Arg(10)-HOE 140 (1 microM) had no effect on bradykinin-stimulated ouabain-sensitive (86)Rb(+) uptake. The B(2) receptor antagonists HOE 140 and WIN 64338 antagonized bradykinin-stimulated ouabain-sensitive (86)Rb(+) uptake with pK(B) values (-log M) of 8.20 +/- 0.08 and 8.11 +/- 0.20 respectively. 5. Reducing extracellular Na+ from 146 mM to 11 mM caused a 53.5% decrease in basal ouabain-sensitive (86)Rb+ uptake and abolished bradykinin-induced uptake. Two inhibitors of the Na(+)-H(+) exchanger, methylisobutyl-amiloride (MIA; 1 - 100 microM) and ethylisopropyl-amiloride (EIPA; 0.1 - 10 microM), inhibited bradykinin-stimulated ouabain-sensitive (86)Rb(+) uptake without affecting basal uptake. 6. These results suggest that bradykinin increases Na+-K+ pump activity of guinea-pig tracheal smooth muscle via stimulation of B(2) receptors and activation of the Na+-H+ exchanger. PMID- 11498521 TI - Human colonic anti-secretory activity of the potent NK(1) antagonist, SR140333: assessment of potential anti-diarrhoeal activity in food allergy and inflammatory bowel disease. AB - 1. This in vitro study was designed to determine the potential use of the NK(1) antagonist, SR140333 as an anti-diarrhoeal treatment for food allergy or inflammatory bowel disease. The effect of various immune and neuronal stimuli on human colonic substance P (SP) release and the effect of SR140333 on subsequently stimulated mucosal ion transport was investigated. 2. Submucosal and sensory nerve fibre stimulation using electrical field stimulation (1 ms/7 Hz/7 V) and capsaicin (50 microM) respectively, mast cell activation by anti-IgE (1/250 dilution) and granulocyte stimulation using fMLP (50 microM) each released SP and evoked a secretory response. 3. SP and the NK(1) selective agonist, Sar-SP (0.1 - 1000 nM) stimulated an increase in colonic secretion which was antagonized by SR140333 (pD'(2)=6.7 and 7.25 versus SP and Sar-SP respectively). 4. SR140333, at a concentration that blocked NK(1)-mediated secretion (500 nM), also reduced the secretory response to both alphaIgE and capsaicin. This suggests a pathophysiologic role for NK(1) receptors. 5. Capsaicin evoked SP release was increased in tissue taken from Crohn's disease but not ulcerative colitis patients. The response to SP was however reduced by 70 and 89% respectively. 6. Mast cells and sensory afferents contribute to allergic diarrhoea. Since SR140333 reduced the secretory response to mast cell and afferent stimulation this compound may be particularly useful in reducing the symptoms of food allergy. PMID- 11498522 TI - Diazepam increases the hypothalamic-pituitary-adrenocortical (HPA) axis activity by a cyclic AMP-dependent mechanism. AB - 1. Previous studies in this laboratory have shown that diazepam behaves as a phosphodiesterase 4 (PDE 4) inhibitor. It has been reported that PDE-4 inhibitors activate the hypothalamic-pituitary-adrenocortical (HPA) axis in the rat. In the present study we have examined whether activation of the cyclic AMP-dependent protein kinase (PKA) is involved in the effect of diazepam on basal HPA axis activity. 2. Acute systemic administration of diazepam (10 mg kg(-1) i.p.) was found to increase the basal HPA axis activity, increasing the plasma concentrations of corticotrophin (ACTH) and corticosterone 30 min post injection. Diazepam also elevated cyclic AMP content of the hypothalamus. 3. Pretreatment of the animals with dexamethasone (1 mg kg(-1) s.c.) for 3 days completely abolished the effect of diazepam on HPA axis activity. 4. The antagonists of central and peripheral benzodiazepine receptors, flumazenil (10 mg kg(-1) i.p.) and PK 11195 (5 mg kg(-1) i.p.) did not affect the diazepam induced increase of HPA axis activity nor did they have an effect per se. 5. The increase in ACTH and corticosterone levels was significantly reduced by the cyclic AMP-dependent protein kinase (PKA) inhibitor, H-89, given either subcutaneously (5 mg kg(-1) s.c.) or intracerebroventricularly (i.c.v.; 28 microg in 10 microl). 6. The results indicate that diazepam can stimulate basal HPA axis activity in the rat by a cyclic AMP-dependent PKA mediated pathway. PMID- 11498523 TI - Acute and chronic effects of desipramine and clorgyline on alpha(2)-adrenoceptors regulating noradrenergic transmission in the rat brain: a dual-probe microdialysis study. AB - 1. The effects of desipramine (3 mg kg(-1) i.p.) and clorgyline (1 mg kg(-1) i.p.) on extracellular noradrenaline (NA) in the locus coeruleus (LC) and cingulate cortex were assessed in freely-moving rats by dual-probe microdialysis. Functional activities of alpha(2)-adrenoceptors regulating NA release in the LC and cingulate cortex were determined by systemic (0.3 mg kg(-1) i.p.) or local (0.1 - 100 microM) clonidine administration. 2. Extracellular NA was increased in the LC and cingulate cortex following acute desipramine but not clorgyline treatment. Systemic clonidine decreased NA similarly in desipramine-, clorgyline , and saline-treated animals, in both brain areas. 3. Long-term (twice daily, 14 days) but not short-term (twice daily, 7 days) desipramine, and long-term clorgyline (once daily, 21 days) treatments increased NA (3 fold) in cingulate cortex but not in the LC. Following long-term treatments, responses of NA to systemic clonidine were attenuated in the LC and cingulate cortex. 4. Clonidine perfusion by reverse dialysis into the cingulate cortex decreased local NA (-55 +/- 9%). The effect was attenuated by long-term desipramine (-31 +/- 9%) and clorgyline (-10 +/- 2%) treatments. 5. Clonidine perfusion by reverse dialysis into the LC decreased NA in the LC (-89 +/- 2%) and in cingulate cortex (-52 +/- 12%). This effect was attenuated in the LC following long-term desipramine (-72 +/- 4%) and clorgyline (-62 +/- 12%) treatments but it was not modified in the cingulate cortex (-57 +/- 10% and -68 +/- 6%, respectively). 6. These findings demonstrate that chronic desipramine or clorgyline treatments increase NA in noradrenergic terminal areas and desensitize alpha(2)-adrenoceptors modulating local NA release at somatodendritic and terminal levels. However, somatodendritic alpha(2)-adrenoceptors that control LC firing activity are not desensitized. PMID- 11498524 TI - Effects of the novel selective endothelin ET(A) receptor antagonist, SB 234551, on the cardiovascular responses to endotoxaemia in conscious rats. AB - 1. In conscious, freely moving, male, Long Evans rats, regional haemodynamic responses to exogenous endothelin-1 (ET-1; 25, 50 and 250 pmol kg(-1) i.v.) were assessed in the presence of vehicle, or the selective ET(A)-receptor antagonist, SB 234551. On the following day, the effects of SB 234551 on the haemodynamic responses to lipopolysaccharide (LPS) infusion (150 microg kg(-1) h(-1), i.v.) were determined. 2. When SB 234551 was given i.v. by primed infusion at a dose of 0.3 mg kg(-1) bolus, 0.3 mg kg(-1) h(-1) infusion, it caused selective inhibition of the vasoconstrictor effects of exogenous endothelin-1, whereas at a dose of 1 mg kg(-1), 1 mg kg(-1) h(-1), SB 234551 also inhibited some of the vasodilator effects of endothelin-1. 3. Infusion of LPS, in the presence of vehicle, caused a short-lived (1 - 2 h) hypotension, tachycardia, and vasodilatation in renal, superior mesenteric and hindquarters vascular beds. Thereafter, blood pressure, heart rate and mesenteric vascular conductance returned to baseline values, but renal vasodilatation persisted, and there was vasoconstriction in the hindquarters. 4. In the presence of SB 234551 (0.3 mg kg(-1), 0.3 mg kg(-1) h( 1)), the early (1 - 2 h) cardiovascular responses to LPS infusion were unaffected, but the subsequent recovery of mean arterial blood pressure was impaired, due to developing vasodilatation in the mesenteric and, to a lesser extent, hindquarters, vascular beds. SB 234551 had no effect on the renal haemodynamic responses to LPS infusion. 5. The results confirm an important, regionally-selective, vasoconstrictor role for endogenous endothelin in this model of endotoxaemia. PMID- 11498525 TI - Activation of the p38 and p42/p44 mitogen-activated protein kinase families by the histamine H(1) receptor in DDT(1)MF-2 cells. AB - 1. The mitogen-activated protein kinases (MAPKs) consist of the p42/p44 MAPKs and the stress-activated protein kinases, c-Jun N-terminal kinase (JNK) and p38 MAPK. In this study we have examined the effect of histamine H(1) receptor activation on MAPK pathway activation in the smooth muscle cell line DDT(1)MF-2. 2. Histamine stimulated time and concentration-dependent increases in p42/p44 MAPK activation in DDT(1)MF-2 cells. Responses to histamine were inhibited by the histamine H(1) receptor antagonist mepyramine (K(D) 3.5 nM) and following pre treatment with pertussis toxin (PTX; 57% inhibition). 3. Histamine-induced increases in p42/p44 MAPK activation were blocked by inhibitors of MAPK kinase 1 (PD 98059), tyrosine kinase (genistein and tyrphostin A47), phosphatidylinositol 3-kinase (wortmannin and LY 294002) and protein kinase C (Ro 31-8220; 10 microM; 41% inhibition). Inhibitors of Src tyrosine kinase (PP2) and the epidermal growth factor tyrosine kinase (AG1478) were without effect. Removal of extracellular Ca(2+), chelation of intracellular Ca(2+) with BAPTA and inhibition of focal adhesion assembly (cytochalasin D) had no significant effect on histamine-induced p42/p44 MAPK activation. 4. Histamine stimulated time and concentration-dependent increases in p38 MAPK activation in DDT(1)MF-2 cells but had no effect on JNK activation. Histamine-induced p38 MAPK activation was inhibited by pertussis toxin (74% inhibition) and the p38 MAPK inhibitor SB 203580 (95% inhibition). 5. In summary, we have shown the histamine H(1) receptor activates p42/p44 MAPK and p38 MAPK signalling pathways in DDT(1)MF-2 smooth muscle cells. Interestingly, signalling to both pathways appears to involve histamine H(1) receptor coupling to G(i)/G(o)-proteins. PMID- 11498526 TI - High level of alpha2-adrenoceptor in rat foetal liver and placenta is due to alpha2B-subtype expression in haematopoietic cells of the erythrocyte lineage. AB - 1. Rat foetal liver contains large amounts of alpha2-adrenoceptors. The present work aimed to identify the receptor subtype and the cell type accounting for high expression and to clarify the mechanisms responsible for the sharp decrease in hepatic receptivity occurring during the late stage of foetal development. 2. Binding experiments indicated that the density of alpha2-adrenoceptors in the foetal liver (embryonic day 18; 615+/-155 fmol mg(-1) of protein) is 18 fold higher than in newborn or adult (35.2+/-4.3 fmol mg(-1)). A high amount of receptor is also found in the placenta (443+/-53 fmol mg(-1)). In both tissues, the rank order of antagonists to inhibit radioligand binding matched the pharmacological profile of the alpha2B-adrenoceptor and exclusively RNG transcripts were detected by RNase protection assays. 3. Isolation of cell fractions from foetal liver showed that alpha2B-adrenoceptor is primarily expressed by haematopoietic cells. Consistent with this view, the receptor is found to be abundant in foetal blood, carried by reticulocytes. The expression in blood gradually declines to zero at 3 weeks of age and it is not recovered following induction of reticulocytosis in adults. 4. In foetal reticulocytes, a low proportion of the receptor population is coupled to G-protein. The alpha2 agonist UK14304 has a marginal effect on cyclic AMP level but significantly increases arachidonic acid release. The function of the receptor remains to be elucidated. However, together with observations on alpha2B-knockout mice, the current finding strongly suggests a role for alpha2B-adrenoceptor during foetal haematopoiesis in rodents. PMID- 11498527 TI - Estimation of anti-platelet drugs on human platelet aggregation with a novel whole blood aggregometer by a screen filtration pressure method. AB - 1. The effects of anti-platelet drugs on human whole blood aggregation were evaluated using a novel whole blood aggregometer by a screen filtration pressure (SFP) method. 2. The SFP whole blood aggregometer was found to successfully detect whole blood aggregation induced by ADP, collagen and TRAP by measuring the SFP of blood samples. The platelet aggregation threshold index (PATI), the concentration of agonist required with an inducing pressure rate of 50%, varied time-dependently after collection of blood. High values for ADP and collagen were noted immediately after blood collection, suggesting low aggregation activity of platelets, and gradually increase thereafter. 3. Cilostazol (phosphodiesterase 3 inhibitor), dipyridamole, aspirin and tirofiban all inhibited whole blood aggregation in vitro. Inhibitory effects of cilostazol and dipyridamole, but not tirofiban, were markedly enhanced 6 or 7 fold by long pre-incubation (60 min), compared with short pre-incubation (2 min). Such enhancement was only observed with ADP- and not collagen-induced whole blood aggregation. A similar phenomenon was also observed for aggregation with platelet rich plasma (PRP). Cilostazol inhibition of ADP-induced platelet aggregation was more potent with PRP than whole blood (PATI(200)=3.80+/-0.95 microM for whole blood; 2.04+/-0.61 microM for PRP). Inhibitory effects of dipyridamole were attenuated in PRP without erythrocytes. 4. These results demonstrate that the SFP aggregometer can sensitively detect anti-platelet aggregatory effects of various kinds of drugs. So that it is a useful tool for evaluation of anti-platelet drugs. PMID- 11498528 TI - CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non peptide antagonist. AB - 1. Calcitonin gene-related peptide (CGRP), amylin and adrenomedullin (AM) belong to the same family of peptides. Accumulating evidence indicate that the calcitonin (CT) receptor, the CT receptor-like receptor (CRLR) and receptor activity-modifying proteins (RAMPs) form the basis of all the receptors in this family of peptides. 2. Using reverse transcriptase - polymerase chain reaction the presence of mRNA sequences encoding the CRLR, RAMP1 and RAMP2 were demonstrated in porcine left anterior descending (LAD) coronary arteries, whereas porcine calcitonin (CT) receptor mRNA was not present. The partial porcine mRNA sequences shared 82 - 92% nucleotide identity with human sequences. 3. The human peptides alphaCGRP, betaCGRP, AM and amylin induced relaxation with pEC(50) values of 8.1, 8.1, 6.7 and 6.1 M respectively. 4. The antagonistic properties of a novel non-peptide CGRP antagonist 'Compound 1' (WO98/11128), betaCGRP(8 - 37) and the proposed AM receptor antagonist AM(22 - 52) were compared to the well known CGRP(1) receptor antagonist alphaCGRP(8 - 37). 5. The alphaCGRP(8 - 37) and betaCGRP(8 - 37) induced concentration-dependent (10(-7) - 10(-5) M) rightward shift of both the alphaCGRP and betaCGRP concentration-response curves. betaCGRP(8 - 37) (10(-6) M) had the same effect as alphaCGRP(8 - 37) (10(-6) M), but with less potent rightward shift of the concentration-response curves for alphaCGRP, AM and amylin. 6. Preincubation with 'Compound 1' (10(-7) - 10(-5) M) and AM(22 - 52) (10(-6) M) had no significant antagonistic effect. 7. In conclusion, the building blocks forming CGRP and AM receptors were present in the porcine LAD, whereas those of the amylin receptor were not. alphaCGRP, betaCGRP, AM and amylin mediated vasorelaxation via the CGRP receptors. No functional response was detected to adrenomedullin via the adrenomedullin receptor. PMID- 11498529 TI - Thalidomide impairment of trinitrobenzene sulphonic acid-induced colitis in the rat - role of endothelial cell-leukocyte interaction. AB - 1. Immune response-modulating drugs such as thalidomide may be of therapeutic value in the treatment of chronic inflammatory bowel diseases including Crohn's disease (CD). In the present study, we have investigated whether thalidomide exerts this effect by impairing endothelial cell-leukocyte interaction through down-regulation of the expression of pro-inflammatory gene products in these cells. 2. Transient CD-like colitis was induced in male Wistar rats by single enema with trinitrobenzene sulphonic acid (TNBS) in ethanol followed by macroscopic scoring, histology, intravital microscopy, RT - PCR and immunohistochemistry (IHC) analyses. Thalidomide or its analogue supidimide were administered in olive oil by intragastric instillation 6 h prior to the induction of colitis and then daily for one week. 3. Both thalidomide and supidimide (200 mg kg(-1) d(-1)) significantly attenuated TNBS-induced colitis as compared to vehicle-treated control animals (44 and 37% inhibition, respectively), and this effect persisted for 7 days post cessation of thalidomide treatment (46% inhibition). 4. Moreover, thalidomide significantly reduced leukocyte sticking to postcapillary venular endothelial cells in the submucosa (by 45%), improved functional capillary density and perfusion, and attenuated endothelial interleukin-8 expression, as judged by IHC analysis. According to RT - PCR analysis, both thalidomide and supidimide also significantly reduced vascular cell adhesion molecule-1 mRNA expression in the affected part of the descending colon. 5. These findings suggest that thalidomide and one of its derivatives impairs CD-like TNBS-induced colitis in the rat by down-regulating endothelial adhesion molecule and chemokine expression and, as a consequence, the interaction of these cells with circulating leukocytes. PMID- 11498530 TI - Suppression of poly (ADP-ribose) polymerase activation by 3-aminobenzamide in a rat model of myocardial infarction: long-term morphological and functional consequences. AB - 1. Recent studies demonstrated that inhibition or genetic inactivation of the enzyme poly (ADP-ribose) polymerase (PARP) is beneficial in myocardial reperfusion injury. PARP activation in the reperfused myocardium has been assumed, but not directly demonstrated. Furthermore, the issue whether pharmacological PARP inhibition affords long-term functional benefit in the reperfused myocardium has not been explored. These questions were addressed in the present study. 2. In a rat model of myocardial ischemia (1 h) and reperfusion (up to 24 h), there was a marked and significant activation of PARP in the ischemic borderzone, as determined by poly(ADP-ribose) (PAR) immunohistochemistry. PAR localized to the nuclei of myocytes and infiltrating mononuclear cells. In the core of the infarction, necrotic tissues and diffuse PAR staining were observed. PARP activation remained markedly detectable 24 h after reperfusion. The PARP inhibitor 3-aminobenzamide (20 mg kg(-1) intraperitoneally 10 min before reperfusion, and every 2 h thereafter for 6 h) markedly reduced the activation of the enzyme in myocytes. 3. 3-aminobenzamide significantly protected against myocardial morphological and functional alterations at 24 h post-reperfusion. Notably, infarct size was reduced, circulating creatine kinase activity was attenuated, and myocardial contractility (dP dt(-1)) was restored by 3-aminobenzamide. 4. Our results demonstrate a significant and prolonged activation of PARP in the reperfused myocardium, localizing to the necrotic area and the ischaemic borderzone. Furthermore, the studies demonstrate that PARP inhibition affords long-term beneficial morphological and functional effects in the reperfused myocardium. These data strengthen the notion that pharmacological PARP inhibition is a viable novel experimental approach for protection against myocardial reperfusion injury. PMID- 11498532 TI - Mutational analysis of the different bulge regions of hepatitis C virus domain II and their influence on internal ribosome entry site translational ability. AB - The hepatitis C virus (HCV) 5'-untranslated region and, in particular, domains II to IV are involved in the internal ribosome entry site (IRES) structure. Recent structural evidence has shown that the function of domain II may be to hold the coding RNA in position until the translational machinery is correctly assembled on the decoding site. However, a comprehensive mutational and functional study concerning the importance of the different RNA regions that compose domain II is not yet available. Therefore, we have taken advantage of the recently proposed secondary structure of domain II to design a series of specific mutants. The bulge regions present in the latest secondary structure prediction of domain II were selectively deleted, and the effects of these mutations on IRES translation efficiency were analyzed. Our results show that the introduction of these mutations can variably affect the degree of HCV translation, causing a moderate to total loss of translation ability that correlates with the severity of changes induced in the RNA secondary structure and degree of p25 ribosomal protein UV cross-linking, but not with the ability of the 40S ribosomal subunit to bind the IRES. These findings support the proposed structural role of domain II in HCV translation. PMID- 11498531 TI - N-methyl-D-aspartate-induced alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) receptor down-regulation involves interaction of the carboxyl terminus of GluR2/3 with Pick1. Ligand-binding studies using Sindbis vectors carrying AMPA receptor decoys. AB - The dynamics of alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) type glutamate receptors, as represented by their exocytosis, endocytosis and cytoskeletal linkage, has often been implicated in N-methyl-d-aspartate (NMDA) dependent synaptic plasticity. To explore the molecular mechanisms underlying the AMPA receptor dynamics, cultured hippocampal neurons were stimulated with 100 microm NMDA, and the biochemical and pharmacological changes in the ligand binding activity of AMPA receptor complexes and its subunits, GluR1 and GluR2/3, were investigated. The NMDA treatment reduced the total amount of bound [(3)H]AMPA on the surface of the neurons but not in their total membrane fraction. This process was mimicked by a protein kinase C activator, phorbol ester, but blocked by an inhibitor of the same kinase, calphostin C. The NMDA induced down-regulation of the ligand binding activity was also reflected by the decreased AMPA-triggered channel activity as well as by the cells' reduced immunoreactivity for GluR1. In parallel, the NMDA treatment markedly altered the interaction between the AMPA receptor subunits and their associating molecule(s); the association of PDZ molecules, including Pick1, with GluR2/3 was enhanced in a protein-kinase-C-dependent manner. Viral expression vectors carrying GluR1 and GluR2 C-terminal decoys, both fused to enhanced green fluorescent protein, were transfected into hippocampal neurons to disrupt their interactions. The overexpression of the C-terminal decoy for GluR2 specifically and significantly blocked the NMDA-triggered reduction in [(3)H]AMPA binding, whereas that for GluR1 had no effects. Co-immunoprecipitation using anti-Pick1 antibodies revealed that the overexpressed GluR2 C-terminal decoy indeed prevented Pick1 from interacting with the endogenous GluR2/3. Therefore, these observations suggest that the NMDA-induced down-regulation of the functional AMPA receptors involves the interaction between GluR2/3 subunits and Pick1. PMID- 11498533 TI - Id-1, ITF-2, and Id-2 comprise a network of helix-loop-helix proteins that regulate mammary epithelial cell proliferation, differentiation, and apoptosis. AB - Mammary epithelial cells proliferate, invade the stroma, differentiate, and die in adult mammals by mechanisms that are poorly understood. We found that Id-1, an inhibitor of basic helix-loop-helix transcription factors, regulates mammary epithelial cell growth, differentiation, and invasion in culture. Here, we show that Id-1 is expressed highly during mammary development in virgin mice and during early pregnancy, when proliferation and invasion are high. During mid pregnancy, Id-1 expression declined to undetectable levels as the epithelium differentiated fully. Surprisingly, Id-1 increased during involution, when the epithelium undergoes extensive apoptosis. To determine whether Id-1 regulates both proliferation and apoptosis, we constitutively expressed Id-1 in mammary epithelial cell cultures. Id-1 stimulated proliferation in sparse cultures but induced apoptosis in dense cultures, which reflect epithelial cell density during early pregnancy and involution, respectively. To understand how Id-1 acts, we screened a yeast two-hybrid library from differentiating mammary epithelial cells and identified ITF-2, a basic helix-loop-helix transcription factor, as an Id-1 interacting protein. Overexpression of ITF-2 significantly reduced Id-1 stimulated proliferation and apoptosis. We show further that, in contrast to Id 1, Id-2 was expressed highly in differentiated mammary epithelial cells in vivo and in culture. In culture, Id-2 antisense transcripts blocked differentiation. Our results suggest that Id-1, ITF-2, and Id-2 comprise a network of interacting molecular switches that govern mammary epithelial cell phenotypes. PMID- 11498534 TI - Interaction between pyrin and the apoptotic speck protein (ASC) modulates ASC induced apoptosis. AB - Patients with familial Mediterranean fever suffer sporadic inflammatory attacks characterized by fever and intense pain (in joints, abdomen, or chest). Pyrin, the product of the MEFV locus, is a cytosolic protein whose function is unknown. Using pyrin as a "bait" to probe a yeast two-hybrid library made from neutrophil cDNA, we isolated apoptotic speck protein containing a caspase recruitment domain (CARD) (ASC), a proapoptotic protein that induces the formation of large cytosolic "specks" in transfected cells. We found that when HeLa cells are transfected with ASC, specks are formed. After co-transfection of cells with ASC plus wild type pyrin, an increase in speck-positive cells is found, and speck positive cells show increased survival. Immunofluorescence studies show that pyrin co-localizes with ASC in specks. Speck localization requires exon 1 of pyrin, but exon 1 alone of pyrin does not result in an increase in the number of specks. Exon 1 of pyrin and exon 1 of ASC show 42% sequence similarity and resemble death domain-related structures in modeling studies. These findings link pyrin to apoptosis pathways and suggest that the modulation of cell survival may be a component of the pathophysiology of familial Mediterranean fever. PMID- 11498535 TI - Rottlerin is a mitochondrial uncoupler that decreases cellular ATP levels and indirectly blocks protein kinase Cdelta tyrosine phosphorylation. AB - Protein kinase Cdelta (PKCdelta) is activated by stimuli that increase its tyrosine phosphorylation, including neurotransmitters that initiate fluid secretion in salivary gland (parotid) epithelial cells. Rottlerin, a compound reported to be a PKCdelta-selective inhibitor, rapidly increased the rate of oxygen consumption (QO2) of parotid acinar cells and PC12 cells. In parotid cells, this was distinct from the effects of the muscarinic receptor ligand carbachol, which promoted a sodium pump-dependent increase in respiration. Rottlerin increased the QO2 of isolated rat liver mitochondria to a level similar to that produced when oxidative phosphorylation was initiated by ADP or when mitochondria were uncoupled by carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP). The effects of rottlerin on mitochondrial QO2 were neither mimicked nor blocked by the PKC inhibitor GF109203X. Rottlerin was not effective in blocking PKCdelta activity in vitro. Exposure of freshly isolated parotid acinar cells to rottlerin and FCCP reduced cellular ATP levels and reduced stimuli-dependent increases in tyrosine phosphorylation of PKCdelta. Neither rottlerin nor FCCP reduced stimuli-dependent PKCdelta tyrosine phosphorylation in RPG1 cells (a salivary ductal line) or PC12 cells, consistent with their dependence on glycolysis rather than oxidative phosphorylation for energy-dependent processes. These results demonstrate that rottlerin directly uncouples mitochondrial respiration from oxidative phosphorylation. Previous studies using rottlerin should be evaluated cautiously. PMID- 11498536 TI - Oligomerization of human Gadd45a protein. AB - Gadd45a is an 18-kDa acidic protein that is induced by genotoxic and certain other cellular stresses. The exact function of this protein is not known. However, there is evidence for its involvement in growth control, maintenance of genomic stability, DNA repair, cell cycle control, and apoptosis. Consistently, Gadd45a has previously been shown to interact in vitro and/or in vivo with a number of proteins playing central roles in these cellular processes: proliferating cell nuclear antigen, p21(Cip1/Waf1), Cdc2-CyclinB complex, MTK1, and histones. Adding to this complexity, we have found that Gadd45a self associates in solution, both in vitro and when expressed in the cell. Moreover, Gadd45a can complex with the two other members of the Gadd45 family of stress induced proteins, human Gadd45b (MyD118) and Gadd45g (CR6). Gel-exclusion chromatography, native gel electrophoretic analysis, enzyme-linked immunosorbent assay, and chemical cross-linking showed that recombinant Gadd45a forms dimeric, trimeric, and tetrameric species in vitro, the dimers being the predominant form. Deletion mutant and peptide scanning analyses suggest that Gadd45a has two self association sites: within N-terminal amino acids 33-61 and within 40 C-terminal amino acids. Despite the low abundance of Gadd45a in the cell, oligomer-forming concentrations can probably be achieved in the foci-like nuclear structures formed by the protein upon overexpression. Evidence for a potential role of Gadd45a self-association in altering DNA accessibility on damaged nucleosomes is presented. PMID- 11498537 TI - Heterologous expression of the transcriptional regulator escargot inhibits megakaryocytic endomitosis. AB - Certain cell types escape the strict mechanisms imposed on the majority of somatic cells to ensure the faithful inheritance of parental DNA content. This is the case in many embryonic tissues and certain adult cells such as mammalian hepatocytes and megakaryocytes. Megakaryocytic endomitosis is characterized by repeated S phases followed by abortive mitoses, resulting in mononucleated polyploid cells. Several cell cycle regulators have been proposed to play an active role in megakaryocytic polyploidization; however, little is known about upstream factors that could control endomitosis. Here we show that ectopic expression of the transcriptional repressor escargot interferes with the establishment of megakaryocytic endomitosis. Phorbol ester-induced polyploidization was inhibited in stably transfected megakaryoblastic HEL cells constitutively expressing escargot. Analysis of the expression and activity of different cell cycle factors revealed that Escargot affects the G(1)/S transition by influencing Cdk2 activity and cyclin A transcription. Nuclear proteins that specifically bind the Escargot-binding element were detected in endomitotic and non-endomitotic megakaryoblastic cells, but down-regulation occurred only during differentiation of cells that become polyploid. As Escargot was originally implicated in ploidy maintenance of Drosophila embryonic and larval cells, our results suggest that polyploidization in megakaryocytes might respond to mechanisms conserved from early development to adult cells that need to escape normal control of the diploid state. PMID- 11498538 TI - Identification and characterization of a synaptojanin 2 splice isoform predominantly expressed in nerve terminals. AB - We have previously identified synaptojanin 1, a phosphoinositide phosphatase predominantly expressed in the nervous system, and synaptojanin 2, a broadly expressed isoform. Synaptojanin 1 is concentrated in nerve terminals, where it has been implicated in synaptic vesicle recycling and actin function. Synaptojanin 2A is targeted to mitochondria via a PDZ domain-mediated interaction. We have now characterized an alternatively spliced form of synaptojanin 2 that shares several properties with synaptojanin 1. This isoform, synaptojanin 2B, undergoes further alternative splicing to generate synaptojanin 2B1 and 2B2. Both amphiphysin and endophilin, two partners synaptojanin 1, bind synaptojanin 2B2, whereas only amphiphysin binds synaptojanin 2B1. Sequence similar to the endophilin-binding site in synaptojanin 1 is present only in synaptojanin 2B2, and this sequence was capable of affinity purifying endophilin from rat brain. The Sac1 domain of synaptojanin 2 exhibited phosphoinositide phosphatase activity very similar to that of the Sac1 domain of synaptojanin 1. Site-directed mutagenesis further illustrated its functional similarity to the catalytic domain of Sac1 proteins. Antibodies raised against the synaptojanin 2B specific carboxyl-terminal region identified a 160-kDa protein in brain and testis. Immunofluorescence showed that synaptojanin 2B is localized at nerve terminals in brain and at the spermatid manchette in testis. Active Rac1 GTPase affects the intracellular localization of synaptojanin 2, but not of synaptojanin 1. These results suggest that synaptojanin 2B has a partially overlapping function with synaptojanin 1 in nerve terminals, with additional roles in neurons and other cells including spermatids. PMID- 11498539 TI - a4, a unique kidney-specific isoform of mouse vacuolar H+-ATPase subunit a. AB - The vacuolar-type H+ -ATPase (V-ATPase) translocates protons across membranes. Here, we have identified a mouse cDNA coding for a fourth isoform (a4) of the membrane sector subunit a of V-ATPase. This isoform was specifically expressed in kidney, but not in the heart, brain, spleen, lung, liver, muscle, or testis. Immunoprecipitation experiments, together with sequence similarities for other isoforms (a1, a2, and a3), indicate that the a4 isoform is a component of V ATPase. Moreover, histochemical studies show that a4 is localized in the apical and basolateral plasma membranes of cortical alpha- and beta-intercalated cells, respectively. These results suggest that the V-ATPase, with the a4 isoform, is important for renal acid/base homeostasis. PMID- 11498540 TI - A small molecule ligand of the glucagon-like peptide 1 receptor targets its amino terminal hormone binding domain. AB - The glucagon-like peptide 1 receptor (GLP-1R) belongs to a distinct subgroup of G protein-coupled peptide hormone receptors (class B) that has been difficult to target by small molecule drugs. Here, we report that a non-peptide compound, T 0632, binds with micromolar affinity to the human GLP-1R and blocks GLP-1-induced cAMP production. Furthermore, the observation that T-0632 has almost 100-fold selectivity for the human versus the highly homologous rat GLP-1R provided an opportunity to map determinants of non-peptide binding. Radioligand competition experiments utilizing a series of chimeric human/rat GLP-1R constructs revealed that partial substitution of the amino terminus of the rat GLP-1R with the corresponding sequence from the human homolog was sufficient to confer high T 0632 affinity. Follow-up analysis of receptors where individual candidate amino acids had been exchanged between the human and rat GLP-1Rs identified a single residue that explained species selectivity of non-peptide binding. Replacement of tryptophan 33 in the human GLP-1R by serine (the homologous amino acid in the rat GLP-1R) resulted in a 100-fold loss of T-0632 affinity, whereas the converse mutation in the rat GLP-1R led to a reciprocal gain-of-function phenotype. These observations suggest that in a class B receptor, important determinants of non peptide affinity reside within the extracellular amino-terminal domain. Compound T-0632 may mimic, and thereby interfere with, the putative "pseudo-tethering" mechanism by which the amino terminus of class B receptors initiates the binding of cognate hormones. PMID- 11498541 TI - Amino acid and insulin signaling via the mTOR/p70 S6 kinase pathway. A negative feedback mechanism leading to insulin resistance in skeletal muscle cells. AB - Amino acids have emerged as potent modulators of the mTOR/p70 S6 kinase pathway. The involvement of this pathway in the regulation of insulin-stimulated glucose transport was investigated in the present study. Acute exposure (1 h) to a balanced mixture of amino acids reduced insulin-stimulated glucose transport by as much as 55% in L6 muscle cells. The effect of amino acids was fully prevented by the specific mTOR inhibitor rapamycin. Time course analysis of insulin receptor substrate 1 (IRS-1)-associated phosphatidylinositol (PI) 3-kinase activity revealed that incubation with amino acids speeds up its time-dependent deactivation, leading to a dramatic suppression (-70%) of its activity after 30 min of insulin stimulation as compared with its maximal activation (5 min of stimulation). This accelerated deactivation of PI 3-kinase activity in amino acid treated cells was associated with a concomitant and sustained increase in the phosphorylation of p70 S6 kinase. In marked contrast, inhibition of mTOR by rapamycin maintained PI 3-kinase maximally activated for up to 30 min. The marked inhibition of insulin-mediated PI 3-kinase activity by amino acids was linked to a rapamycin-sensitive increase in serine/threonine phosphorylation of IRS-1 and a decreased binding of the p85 subunit of PI 3-kinase to IRS-1. Furthermore, amino acids were required for the degradation of IRS-1 during long term insulin treatment. These results identify the mTOR/p70 S6 kinase signaling pathway as a novel modulator of insulin-stimulated glucose transport in skeletal muscle cells. PMID- 11498542 TI - Follicle-stimulating hormone stimulates protein kinase A-mediated histone H3 phosphorylation and acetylation leading to select gene activation in ovarian granulosa cells. AB - We examined the phosphorylation and acetylation of histone H3 in ovarian granulosa cells stimulated to differentiate by follicle-stimulating hormone (FSH). We found that protein kinase A (PKA) mediates H3 phosphorylation on serine 10, based on inhibition exclusively by PKA inhibitors. FSH-stimulated H3 phosphorylation in granulosa cells is not downstream of mitogen-activated protein kinase/extracellular signal-regulated kinase, ribosomal S6 kinase-2, mitogen- and stress-activated protein kinase-1, p38 MAPK, phosphatidylinositol-3 kinase, or protein kinase C. Transcriptional activation-associated H3 phosphorylation on serine 10 and acetylation of lysine 14 leads to activation of serum glucocorticoid kinase, inhibin alpha, and c-fos genes. We propose that phosphorylation of histone H3 on serine 10 by PKA in coordination with acetylation of H3 on lysine 14 results in reorganization of the promoters of select FSH responsive genes into a more accessible configuration for activation. The unique role of PKA as the physiological histone H3 kinase is consistent with the central role of PKA in initiating granulosa cell differentiation. PMID- 11498543 TI - Morphine metabolism in the opium poppy and its possible physiological function. Biochemical characterization of the morphine metabolite, bismorphine. AB - We identified a novel metabolic system of morphine in the opium poppy (Papaver somniferum L.). In response to stress, morphine is quickly metabolized to bismorphine consisting of two morphine units, followed by accumulation in the cell wall. This bismorphine binds predominantly to pectins, which possess high galacturonic acid residue contents, through ionical bonds. Our newly developed method using artificial polysaccharides demonstrated that bismorphine bridges are formed between the two amino groups of bismorphine and the carboxyl groups of galacturonic acid residues, resulting in cross-linking of galacturonic acid containing polysaccharides to each other. The ability of bismorphine to cross link pectins is much higher than that of Ca2+, which also acts as a cross-linker of these polysaccharides. Furthermore, we confirmed that cross-linking of pectins through bismorphine bridges leads to resistance against hydrolysis by pectinases. These results indicated that production of bismorphine is a defense response of the opium poppy. Bismorphine formation is catalyzed by anionic peroxidase that pre-exists in the capsules and leaves of opium poppies. The constitutive presence of morphine, together with bismorphine-forming peroxidase, enables the opium poppy to rapidly induce the defense system. PMID- 11498544 TI - Loss of caveolae, vascular dysfunction, and pulmonary defects in caveolin-1 gene disrupted mice. AB - Caveolae are plasma membrane invaginations that may play an important role in numerous cellular processes including transport, signaling, and tumor suppression. By targeted disruption of caveolin-1, the main protein component of caveolae, we generated mice that lacked caveolae. The absence of this organelle impaired nitric oxide and calcium signaling in the cardiovascular system, causing aberrations in endothelium-dependent relaxation, contractility, and maintenance of myogenic tone. In addition, the lungs of knockout animals displayed thickening of alveolar septa caused by uncontrolled endothelial cell proliferation and fibrosis, resulting in severe physical limitations in caveolin-1-disrupted mice. Thus, caveolin-1 and caveolae play a fundamental role in organizing multiple signaling pathways in the cell. PMID- 11498545 TI - Origin of the hard x-ray emission from the Galactic plane. AB - The Galactic plane is a strong emitter of hard x-rays (2 to 10 kiloelectron volts), and the emission forms a narrow continuous ridge. The currently known hard x-ray sources are far too few to explain the ridge x-ray emission, and the fundamental question of whether the ridge emission is ultimately resolved into numerous dimmer discrete sources or truly diffuse emission has not yet been settled. In order to obtain a decisive answer, using the Chandra X-ray Observatory, we carried out the deepest hard x-ray survey of a Galactic plane region that is devoid of known x-ray point sources. We detected at least 36 new hard x-ray point sources in addition to strong diffuse emission within a 17' by 17' field of view. The surface density of the point sources is comparable to that at high Galactic latitudes after the effects of Galactic absorption are considered. Therefore, most of these point sources are probably extragalactic, presumably active galaxies seen through the Galactic disk. The Galactic ridge hard x-ray emission is diffuse, which indicates omnipresence within the Galactic plane of a hot plasma, the energy density of which is more than one order of magnitude higher than any other substance in the interstellar space. PMID- 11498547 TI - The effectiveness of parks. PMID- 11498546 TI - Correlation between histone lysine methylation and developmental changes at the chicken beta-globin locus. AB - Methylation of histones at specific residues plays an important role in transcriptional regulation. Chromatin immunoprecipitation of dimethylated lysine 9 on histone H3 across 53 kilobases of the chicken beta-globin locus during erythropoiesis shows an almost complete anticorrelation between regions of elevated lysine 9 methylation and acetylation. Lysine 9 is methylated most over constitutive condensed chromatin and developmentally inactive globin genes. In contrast, lysine 4 methylation of histone H3 correlates with H3 acetylation. These results lead us to propose a mechanism by which the insulator in the beta globin locus can protect the globin genes from being silenced by adjacent condensed chromatin. PMID- 11498548 TI - Death at Johns Hopkins. PMID- 11498549 TI - Clinical research. Hopkins reviews investment in Indian cancer drug trial. PMID- 11498551 TI - Russian science. Academy backs off cold war-style rules. PMID- 11498550 TI - U.S. Congress. Would cloning ban affect stem cells? PMID- 11498552 TI - Evolutionary biology. Molecular approach to mushroom hunting. PMID- 11498553 TI - Infectious diseases. Sand fly saliva may be key to new vaccine. PMID- 11498554 TI - Fighting brain drain. Ireland gives its stars a big pot o' gold. PMID- 11498555 TI - Hypertension. Possible new path for blood pressure control. PMID- 11498557 TI - 2002 budget. Fall fight looms over space science funding. PMID- 11498556 TI - Astrophysics. Smooth x-rays fill the Milky Way's disk. PMID- 11498558 TI - Marine science. Mideast pirates give oceanographers pause. PMID- 11498559 TI - Intellectual property. Riken scientist quits; lab says it's clean. PMID- 11498560 TI - Building a small-animal model for AIDS, block by block. PMID- 11498561 TI - Paleontology. Mass extinctions face downsizing, extinction. PMID- 11498563 TI - Profile. Twin stars of astrophysics make room for two. PMID- 11498562 TI - China. Biochemist wages online war against ethical lapses. PMID- 11498564 TI - Gene regulation. A paradigm for precision. PMID- 11498565 TI - Cosmology. Probing matter at the lowest densities. PMID- 11498566 TI - Isotope geochemistry. The origin of water on earth. PMID- 11498568 TI - Solar energy. Solar cells by self-assembly? PMID- 11498567 TI - Protein synthesis. Believe it or not-translation in the nucleus. PMID- 11498569 TI - The evolution of epigenetics. PMID- 11498570 TI - Behind the scenes of gene expression. PMID- 11498571 TI - Champion of chromatin. Alan Wolffe (1959-2001). PMID- 11498572 TI - RNA rules--at least sometimes. PMID- 11498573 TI - The role of DNA methylation in mammalian epigenetics. AB - Genes constitute only a small proportion of the total mammalian genome, and the precise control of their expression in the presence of an overwhelming background of noncoding DNA presents a substantial problem for their regulation. Noncoding DNA, containing introns, repetitive elements, and potentially active transposable elements, requires effective mechanisms for its long-term silencing. Mammals appear to have taken advantage of the possibilities afforded by cytosine methylation to provide a heritable mechanism for altering DNA-protein interactions to assist in such silencing. Genes can be transcribed from methylation-free promoters even though adjacent transcribed and nontranscribed regions are extensively methylated. Gene promoters can be used and regulated while keeping noncoding DNA, including transposable elements, suppressed. Methylation is also used for long-term epigenetic silencing of X-linked and imprinted genes and can either increase or decrease the level of transcription, depending on whether the methylation inactivates a positive or negative regulatory element. PMID- 11498574 TI - DNA methylation and epigenetic inheritance in plants and filamentous fungi. AB - Plants and filamentous fungi share with mammals enzymes responsible for DNA methylation. In these organisms, DNA methylation is associated with gene silencing and transposon control. However, plants and fungi differ from mammals in the genomic distribution, sequence specificity, and heritability of methylation. We consider the role that transposons play in establishing methylation patterns and the epigenetic consequences of their perturbation. PMID- 11498575 TI - Translating the histone code. AB - Chromatin, the physiological template of all eukaryotic genetic information, is subject to a diverse array of posttranslational modifications that largely impinge on histone amino termini, thereby regulating access to the underlying DNA. Distinct histone amino-terminal modifications can generate synergistic or antagonistic interaction affinities for chromatin-associated proteins, which in turn dictate dynamic transitions between transcriptionally active or transcriptionally silent chromatin states. The combinatorial nature of histone amino-terminal modifications thus reveals a "histone code" that considerably extends the information potential of the genetic code. We propose that this epigenetic marking system represents a fundamental regulatory mechanism that has an impact on most, if not all, chromatin-templated processes, with far-reaching consequences for cell fate decisions and both normal and pathological development. PMID- 11498576 TI - RNA: guiding gene silencing. AB - In diverse organisms, small RNAs derived from cleavage of double-stranded RNA can trigger epigenetic gene silencing in the cytoplasm and at the genome level. Small RNAs can guide posttranscriptional degradation of complementary messenger RNAs and, in plants, transcriptional gene silencing by methylation of homologous DNA sequences. RNA silencing is a potent means to counteract foreign sequences and could play an important role in plant and animal development. PMID- 11498577 TI - Epigenetic aspects of X-chromosome dosage compensation. AB - The X chromosomes of mammals and fruit flies exhibit unusual properties that have evolved to deal with the different dosages of X-linked genes in males (XY) and females (XX). The X chromosome dosage-compensation mechanisms discovered in these species are evolutionarily unrelated, but exhibit surprising parallels in their regulatory strategies. These features include the importance of noncoding RNAs, and epigenetic spreading of chromatin-modifying activities. Sex chromosomes have posed a fascinating puzzle for biologists. The dissimilar organization, gene content, and regulation of the X and Y chromosomes are thought to reflect selective forces acting on original pairs of identical chromosomes (1-3). The result in many organisms is a male-specific Y chromosome that has lost most of its original genetic content, and a difference in dosage of the X chromosome in males (XY) and females (XX). PMID- 11498578 TI - Imprinting and the epigenetic asymmetry between parental genomes. AB - Genomic imprinting confers a developmental asymmetry on the parental genomes, through epigenetic modifications in the germ line and embryo. These heritable modifications regulate the monoallelic activity of parental alleles resulting in their functional differences during development. Specific cis-acting regulatory elements associated with imprinted genes carry modifications involving chromatin structural changes and DNA methylation. Some of these modifications are initiated in the germ line. Comparative genomic analysis at imprinted domains is emerging as a powerful tool for the identification of conserved elements amenable to more detailed functional analysis, and for providing insight into the emergence of imprinting during the evolution of mammalian species. Genomic imprinting therefore provides a model system for the analysis of the epigenetic control of genome function. PMID- 11498579 TI - Epigenetic reprogramming in mammalian development. AB - DNA methylation is a major epigenetic modification of the genome that regulates crucial aspects of its function. Genomic methylation patterns in somatic differentiated cells are generally stable and heritable. However, in mammals there are at least two developmental periods-in germ cells and in preimplantation embryos-in which methylation patterns are reprogrammed genome wide, generating cells with a broad developmental potential. Epigenetic reprogramming in germ cells is critical for imprinting; reprogramming in early embryos also affects imprinting. Reprogramming is likely to have a crucial role in establishing nuclear totipotency in normal development and in cloned animals, and in the erasure of acquired epigenetic information. A role of reprogramming in stem cell differentiation is also envisaged. DNA methylation is one of the best-studied epigenetic modifications of DNA in all unicellular and multicellular organisms. In mammals and other vertebrates, methylation occurs predominantly at the symmetrical dinucleotide CpG (1-4). Symmetrical methylation and the discovery of a DNA methyltransferase that prefers a hemimethylated substrate, Dnmt1 (4), suggested a mechanism by which specific patterns of methylation in the genome could be maintained. Patterns imposed on the genome at defined developmental time points in precursor cells could be maintained by Dnmt1, and would lead to predetermined programs of gene expression during development in descendants of the precursor cells (5, 6). This provided a means to explain how patterns of differentiation could be maintained by populations of cells. In addition, specific demethylation events in differentiated tissues could then lead to further changes in gene expression as needed. Neat and convincing as this model is, it is still largely unsubstantiated. While effects of methylation on expression of specific genes, particularly imprinted ones (7) and some retrotransposons (8), have been demonstrated in vivo, it is still unclear whether or not methylation is involved in the control of gene expression during normal development (9-13). Although enzymes have been identified that can methylate DNA de novo (Dnmt3a and Dnmt3b) (14), it is unknown how specific patterns of methylation are established in the genome. Mechanisms for active demethylation have been suggested, but no enzymes have been identified that carry out this function in vivo (15-17). Genomewide alterations in methylation-brought about, for example, by knockouts of the methylase genes-result in embryo lethality or developmental defects, but the basis for abnormal development still remains to be discovered (7, 14). What is clear, however, is that in mammals there are developmental periods of genomewide reprogramming of methylation patterns in vivo. Typically, a substantial part of the genome is demethylated, and after some time remethylated, in a cell- or tissue-specific pattern. The developmental dynamics of these reprogramming events, as well as some of the enzymatic mechanisms involved and the biological purposes, are beginning to be understood. Here we look at what is known about reprogramming in mammals and discuss how it might relate to developmental potency and imprinting. PMID- 11498580 TI - Nuclear cloning and epigenetic reprogramming of the genome. AB - Cloning of mammals by nuclear transfer (NT) results in gestational or neonatal failure with at most a few percent of manipulated embryos resulting in live births. Many of those that survive to term succumb to a variety of abnormalities that are likely due to inappropriate epigenetic reprogramming. Cloned embryos derived from donors, such as embryonic stem cells, that may require little or no reprogramming of early developmental genes develop substantially better beyond implantation than NT clones derived from somatic cells. Although recent experiments have demonstrated normal reprogramming of telomere length and X chromosome inactivation, epigenetic information established during gametogenesis, such as gametic imprints, cannot be restored after nuclear transfer. Survival of cloned animals to birth and beyond, despite substantial transcriptional dysregulation, is consistent with mammalian development being rather tolerant to epigenetic abnormalities, with lethality resulting only beyond a threshold of faulty gene reprogramming encompassing multiple loci. PMID- 11498581 TI - The centromere paradox: stable inheritance with rapidly evolving DNA. AB - Every eukaryotic chromosome has a centromere, the locus responsible for poleward movement at mitosis and meiosis. Although conventional loci are specified by their DNA sequences, current evidence favors a chromatin-based inheritance mechanism for centromeres. The chromosome segregation machinery is highly conserved across all eukaryotes, but the DNA and protein components specific to centromeric chromatin are evolving rapidly. Incompatibilities between rapidly evolving centromeric components may be responsible for both the organization of centromeric regions and the reproductive isolation of emerging species. PMID- 11498582 TI - Genes, genetics, and epigenetics: a correspondence. AB - Over the past months, as this special issue took shape, the Editors of Science have monitored an exchange of seven letters initiated by three queries from M. Bacon. These queries concern the popular definitions of "genes," "genetics," and "epigenetics." Below, we reprint excerpts from these letters, referring interested readers to www.sciencemag.org/cgi/content/full/293/5532/1103/DC1 for the complete text and additional references. PMID- 11498583 TI - Human hypertension caused by mutations in WNK kinases. AB - Hypertension is a major public health problem of largely unknown cause. Here, we identify two genes causing pseudohypoaldosteronism type II, a Mendelian trait featuring hypertension, increased renal salt reabsorption, and impaired K+ and H+ excretion. Both genes encode members of the WNK family of serine-threonine kinases. Disease-causing mutations in WNK1 are large intronic deletions that increase WNK1 expression. The mutations in WNK4 are missense, which cluster in a short, highly conserved segment of the encoded protein. Both proteins localize to the distal nephron, a kidney segment involved in salt, K+, and pH homeostasis. WNK1 is cytoplasmic, whereas WNK4 localizes to tight junctions. The WNK kinases and their associated signaling pathway(s) may offer new targets for the development of antihypertensive drugs. PMID- 11498584 TI - Resolving the structure of ionized helium in the intergalactic medium with the far ultraviolet spectroscopic explorer. AB - The neutral hydrogen (H I) and ionized helium (He II) absorption in the spectra of quasars are unique probes of structure in the early universe. We present Far Ultraviolet Spectroscopic Explorer observations of the line of sight to the quasar HE2347-4342 in the 1000 to 1187 angstrom band at a resolving power of 15,000. We resolve the He II Lyman alpha (Lyalpha) absorption as a discrete forest of absorption lines in the redshift range 2.3 to 2.7. About 50 percent of these features have H I counterparts with column densities N(H I) > 10(12.3) per square centimeter that account for most of the observed opacity in He II Lyalpha. The He II to H I column density ratio ranges from 1 to >1000, with an average of approximately 80. Ratios of <100 are consistent with photoionization of the absorbing gas by a hard ionizing spectrum resulting from the integrated light of quasars, but ratios of >100 in many locations indicate additional contributions from starburst galaxies or heavily filtered quasar radiation. The presence of He II Lyalpha absorbers with no H I counterparts indicates that structure is present even in low-density regions, consistent with theoretical predictions of structure formation through gravitational instability. PMID- 11498585 TI - Self-organized discotic liquid crystals for high-efficiency organic photovoltaics. AB - Self-organization of liquid crystalline and crystalline-conjugated materials has been used to create, directly from solution, thin films with structures optimized for use in photodiodes. The discotic liquid crystal hexa-peri-hexabenzocoronene was used in combination with a perylene dye to produce thin films with vertically segregated perylene and hexabenzocoronene, with large interfacial surface area. When incorporated into diode structures, these films show photovoltaic response with external quantum efficiencies of more than 34 percent near 490 nanometers. These efficiencies result from efficient photoinduced charge transfer between the hexabenzocoronene and perylene, as well as from effective transport of charges through vertically segregated perylene and hexabenzocoronene pi systems. This development demonstrates that complex structures can be engineered from novel materials by means of simple solution-processing steps and may enable inexpensive, high-performance, thin-film photovoltaic technology. PMID- 11498586 TI - Polarization mode control of two-dimensional photonic crystal laser by unit cell structure design. AB - We demonstrate polarization mode selection in a two-dimensional (2D) photonic crystal laser by controlling the geometry of the unit cell structure. As the band diagram of the square-lattice photonic crystal is influenced by the unit cell structure, calculations reveal that changing the structure from a circular to an elliptical geometry should result in a strong modification of the electromagnetic field distributions at the band edges. Such a structural modification is expected to provide a mechanism for controlling the polarization modes of the emitted light. A square-lattice photonic crystal with the elliptical unit cell structure has been fabricated and integrated with a gain media. The observed coherent 2D lasing action with a single wavelength and controlled polarization is in good agreement with the predicted behavior. PMID- 11498587 TI - High-temperature ferromagnetism in CaB2C2. AB - We report a high Curie-temperature ferromagnet, CaB2C2. Although the compound has neither transition metal nor rare earth ions, the ferromagnetic transition temperature Tc is about 770 Kelvin. Despite this high T(c), the magnitude of the ordered moment at room temperatures is on the order of 10(-4) Bohr magneton per formula unit. These properties are rather similar to those of doped divalent hexaborides, such as Ca(1-x)La(x)B6. The calculated electronic states also show similarity near the Fermi level between CaB2C2 and divalent hexaborides. However, there is an important difference: CaB2C2 crystallizes in a tetragonal structure, and there are no equivalent pockets in the energy bands for electrons and holes in contrast with CaB6. Thus, the disputed threefold degeneracy, specific to the cubic structure, in the energy bands of divalent hexaborides turns out not to be essential for high-temperature ferromagnetism. It is the peculiar molecular orbitals near the Fermi level that appear to be crucial to the high-Tc ferromagnetism. PMID- 11498588 TI - Runaway growth of planetary embryos facilitated by massive bodies in a protoplanetary disk. AB - About 30% of detected extrasolar planets exist in multiple-star systems. The standard model of planet formation cannot easily accommodate such systems and has difficulty explaining the odd orbital characteristics of most extrasolar giant planets. We demonstrate that the formation of terrestrial-size planets may be insulated from these problems, enabling much of the framework of the standard model to be salvaged for use in complex systems. A type of runaway growth is identified that allows planetary embryos to form by a combination of nebular gas drag and perturbations from massive companions-be they giant planets, brown dwarfs, or other stars. PMID- 11498589 TI - Molecular evidence for the early colonization of land by fungi and plants. AB - The colonization of land by eukaryotes probably was facilitated by a partnership (symbiosis) between a photosynthesizing organism (phototroph) and a fungus. However, the time when colonization occurred remains speculative. The first fossil land plants and fungi appeared 480 to 460 million years ago (Ma), whereas molecular clock estimates suggest an earlier colonization of land, about 600 Ma. Our protein sequence analyses indicate that green algae and major lineages of fungi were present 1000 Ma and that land plants appeared by 700 Ma, possibly affecting Earth's atmosphere, climate, and evolution of animals in the Precambrian. PMID- 11498590 TI - Coordination of a transcriptional switch by HMGI(Y) acetylation. AB - Dynamic control of interferon-beta (IFN-beta) gene expression requires the regulated assembly and disassembly of the enhanceosome, a higher-order nucleoprotein complex formed in response to virus infection. The enhanceosome activates transcription by recruiting the histone acetyltransferase proteins CREB binding protein (CBP) and p300/CBP-associated factors (PCAF)/GCN5, which, in addition to modifying histones, acetylate HMGI(Y), the architectural component required for enhanceosome assembly. We show that the accurate execution of the IFN-beta transcriptional switch depends on the ordered acetylation of the high mobility group I protein HMGI(Y) by PCAF/GCN5 and CBP, which acetylate HMGI(Y) at distinct lysine residues on endogenous promoters. Whereas acetylation of HMGI(Y) by CBP at lysine-65 destabilizes the enhanceosome, acetylation of HMGI(Y) by PCAF/GCN5 at lysine-71 potentiates transcription by stabilizing the enhanceosome and preventing acetylation by CBP. PMID- 11498591 TI - Cytokine-specific transcriptional regulation through an IL-5Ralpha interacting protein. AB - Cytokine receptors consist of multiple subunits, which are often shared between different receptors, resulting in the functional redundancy sometimes observed between cytokines. The interleukin 5 (IL-5) receptor consists of an IL-5-specific alpha-subunit (IL-5Ralpha) and a signal-transducing beta-subunit (betac) shared with the IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors. In this study, we sought to find a role for the cytoplasmic domain of IL-5Ralpha. We show that syntenin, a protein containing PSD-95/Discs large/zO-1 (PDZ) domains, associates with the cytoplasmic tail of the IL-5Ralpha. Syntenin was found to directly associate with the transcription factor Sox4. Association of syntenin with IL-5Ralpha was required for IL-5-mediated activation of Sox4. These studies identify a mechanism of transcriptional activation by cytokine specific receptor subunits. PMID- 11498592 TI - Snf1--a histone kinase that works in concert with the histone acetyltransferase Gcn5 to regulate transcription. AB - Modification of histones is an important element in the regulation of gene expression. Previous work suggested a link between acetylation and phosphorylation, but questioned its mechanistic basis. We have purified a histone H3 serine-10 kinase complex from Saccharomyces cerevisiae and have identified its catalytic subunit as Snf1. The Snf1/AMPK family of kinases function in conserved signal transduction pathways. Our results show that Snf1 and the acetyltransferase Gcn5 function in an obligate sequence to enhance INO1 transcription by modifying histone H3 serine-10 and lysine-14. Thus, phosphorylation and acetylation are targeted to the same histone by promoter specific regulation by a kinase/acetyltransferase pair, supporting models of gene regulation wherein transcription is controlled by coordinated patterns of histone modification. PMID- 11498593 TI - Argonaute2, a link between genetic and biochemical analyses of RNAi. AB - Double-stranded RNA induces potent and specific gene silencing through a process referred to as RNA interference (RNAi) or posttranscriptional gene silencing (PTGS). RNAi is mediated by RNA-induced silencing complex (RISC), a sequence specific, multicomponent nuclease that destroys messenger RNAs homologous to the silencing trigger. RISC is known to contain short RNAs ( approximately 22 nucleotides) derived from the double-stranded RNA trigger, but the protein components of this activity are unknown. Here, we report the biochemical purification of the RNAi effector nuclease from cultured Drosophila cells. The active fraction contains a ribonucleoprotein complex of approximately 500 kilodaltons. Protein microsequencing reveals that one constituent of this complex is a member of the Argonaute family of proteins, which are essential for gene silencing in Caenorhabditis elegans, Neurospora, and Arabidopsis. This observation begins the process of forging links between genetic analysis of RNAi from diverse organisms and the biochemical model of RNAi that is emerging from Drosophila in vitro systems. PMID- 11498594 TI - Transitions in distinct histone H3 methylation patterns at the heterochromatin domain boundaries. AB - Eukaryotic genomes are organized into discrete structural and functional chromatin domains. Here, we show that distinct site-specific histone H3 methylation patterns define euchromatic and heterochromatic chromosomal domains within a 47-kilobase region of the mating-type locus in fission yeast. H3 methylated at lysine 9 (H3 Lys9), and its interacting Swi6 protein, are strictly localized to a 20-kilobase silent heterochromatic interval. In contrast, H3 methylated at lysine 4 (H3 Lys4) is specific to the surrounding euchromatic regions. Two inverted repeats flanking the silent interval serve as boundary elements to mark the borders between heterochromatin and euchromatin. Deletions of these boundary elements lead to spreading of H3 Lys9 methylation and Swi6 into neighboring sequences. Furthermore, the H3 Lys9 methylation and corresponding heterochromatin-associated complexes prevent H3 Lys4 methylation in the silent domain. PMID- 11498595 TI - Crystal structure of a neutralizing human IGG against HIV-1: a template for vaccine design. AB - We present the crystal structure at 2.7 angstrom resolution of the human antibody IgG1 b12. Antibody b12 recognizes the CD4-binding site of human immunodeficiency virus-1 (HIV-1) gp120 and is one of only two known antibodies against gp120 capable of broad and potent neutralization of primary HIV-1 isolates. A key feature of the antibody-combining site is the protruding, finger-like long CDR H3 that can penetrate the recessed CD4-binding site of gp120. A docking model of b12 and gp120 reveals severe structural constraints that explain the extraordinary challenge in eliciting effective neutralizing antibodies similar to b12. The structure, together with mutagenesis studies, provides a rationale for the extensive cross-reactivity of b12 and a valuable framework for the design of HIV 1 vaccines capable of eliciting b12-like activity. PMID- 11498596 TI - Enforcement of temporal fidelity in pyramidal cells by somatic feed-forward inhibition. AB - The temporal resolution of neuronal integration depends on the time window within which excitatory inputs summate to reach the threshold for spike generation. Here, we show that in rat hippocampal pyramidal cells this window is very narrow (less than 2 milliseconds). This narrowness results from the short delay with which disynaptic feed-forward inhibition follows monosynaptic excitation. Simultaneous somatic and dendritic recordings indicate that feed-forward inhibition is much stronger in the soma than in the dendrites, resulting in a broader integration window in the latter compartment. Thus, the subcellular partitioning of feed-forward inhibition enforces precise coincidence detection in the soma, while allowing dendrites to sum incoming activity over broader time windows. PMID- 11498598 TI - A comparison of the X-ray single-crystal structure of Li3SbO4 with the Rietveld refinement. AB - The X-ray single-crystal structure of trilithium antimony tetraoxide, Li(3)SbO(4), is compared with the Rietveld refinement previously reported for the same material. An analysis of the geometric parameters and s.u.'s extracted from both refinements shows that, as expected, powder data yield a less accurate structure. Nevertheless, both refinements give correct geometric parameters within s.u.'s characteristic of each technique. PMID- 11498597 TI - Expectation and dopamine release: mechanism of the placebo effect in Parkinson's disease. AB - The power of placebos has long been recognized for improving numerous medical conditions such as Parkinson's disease (PD). Little is known, however, about the mechanism underlying the placebo effect. Using the ability of endogenous dopamine to compete for [11C]raclopride binding as measured by positron emission tomography, we provide in vivo evidence for substantial release of endogenous dopamine in the striatum of PD patients in response to placebo. Our findings indicate that the placebo effect in PD is powerful and is mediated through activation of the damaged nigrostriatal dopamine system. PMID- 11498599 TI - Na3Co(NO2)6 at 293 and 10 K. AB - The crystal structure of trisodium hexanitrocobaltate(III) has been determined by X-ray diffraction at 293 and 10 K. It contains the slightly distorted octahedral Co(NO(2))(6)(3-) anion. The accurate and extensive data sets collected should be suitable for charge-density analysis studies. PMID- 11498600 TI - Di-mu-phenylthio-bis[bis(eta5-methyl-cyclopentadienyl)(tetrahydrofuran) lanthanum(III)] bis(tetrahydrofuran) solvate. AB - The title complex, [La(2)(C(6)H(7))(4)(C(6)H(5)S)(2)(C(4)H(8)O)(2)].2C(4)H(8)O, is a centrosymmetric dimer bridged through the S atoms of the benzenethiolate ligands. The bridging La(2)S(2) unit is completely planar, while the geometry around the nine-coordinate La atom is that of a distorted trigonal bipyramid. The La-S-La and S-La-S angles are 117.51 (4) and 62.5 (1) degrees, respectively, and the average La-S bond length is 2.9759 A. The crystals contain two tetrahydrofuran solvate molecules for every complex molecule. PMID- 11498601 TI - catena-Poly[aquacopper(II)-mu-[N-(1-oxido-2-naphthylmethylene)-glycinato O,N,O':O']]. AB - In the title compound, [Cu(C(13)H(9)NO(3))(H(2)O)](n), the Cu(II) ion is in a slightly distorted square-pyramidal environment, with four short bonds in the basal plane formed by three donor atoms of the Schiff base and a water O atom. A symmetry-related neighbouring molecule provides an apical carboxylate O atom at a distance of 2.551 (3) A; this contact leads to the formation of zigzag polymeric chains. In addition, the chain fragments are connected to each other by hydrogen bonding. PMID- 11498602 TI - [H3N(CH2)2NH3]0.5[BePO4], a beryllophosphate analogue of aluminosilicate zeolite gismondine. AB - Hydrothermally prepared ethylenediammonium beryllophosphate, (C(2)H(10)N(2))(0.5)[BePO(4)], is an analogue of aluminosilicate zeolite gismondine. A three-dimensional network of vertex-sharing BeO(4) and PO(4) tetrahedra [d(av)(Be-O) = 1.618 (3) A, d(av)(P-O) = 1.5246 (14) A and straight theta(av)(Be-O-P) = 139.8 degrees] encapsulates the disordered ethylenediammonium cations in an eight-ring channel system. PMID- 11498604 TI - (Nitro-kappaN)[2-(phenyldiazenyl-kappaN2)-pyridine-kappaN](2,2':6',2'' terpyridine-kappa3N)ruthenium(II) tetrafluoroborate. AB - The Ru-N bond distances in the title complex, [Ru(NO(2))(C(11)H(9)N(3))(C(15)H(11)N(3))]BF(4) or [Ru(NO(2))(tpy)(azpy)]BF(4), [tpy is 2,2':6',2"-terpyridine and azpy is 2-(phenylazo)pyridine], are Ru-N(py) 2.063 (4), Ru-N(azo) 2.036 (4), Ru-N(nitro) 2.066 (3) A, and Ru-N(tpy) 2.082 (4), 1.982 (3) and 2.074 (4) A. The azo N atom is trans to the nitro group. The azo N=N bond length is 1.265 (5) A, which is the shortest found in such complexes to date. This indicates a multiple bond between Ru and the N atom of the nitro group, and pi-backbonding [dpi(Ru) --> pi*(azo)] is decreased. PMID- 11498603 TI - Bis[mu-2-[(1,5-diaza-1-cyclooctyl)-methyl]phenolato-N,N',O:O]bis [chlorozinc(II)]diacetone solvate: design of a square-pyramidal ZnN2O2Cl complex. AB - The title complex, [Zn(2)(C(13)H(19)N(2)O)(2)Cl(2)].2C(3)H(6)O, resides on a crystallographic inversion center. The two Zn(II) centers bridged by the phenoxo groups are in pentacoordinated distorted square-pyramidal coordination environments with an intramolecular Zn...Zn distance of 3.175 (3) A. The mesocyclic ligand takes a boat-chair conformation and an H atom from the 1,5 diazacyclooctane ring effectively blocks the axial coordination site opposite the Cl ligand to form the ZnN(2)O(2)Cl geometry. The crystal structure is stabilized by a N-H...O hydrogen bond between the amino group and an acetone molecule. PMID- 11498605 TI - Three monoalkoxy-substituted nido-platinaboranes: [(PPh3)2PtB10H11-8-(OCH3)], [(PPh3)2PtB10H11-8-[OCH-(CH3)2]] and [(PPh3)2PtB10H10-9-[OCH(CH3)2]]. AB - Each of the title compounds, 8-methoxy-7,7-bis(triphenylphosphine-P)-8,9:10,11-di muH-7-platina-nido-undecaborane dichloromethane hemisolvate, [Pt(CH(14)B(10)O)(C(18)H(15)P)(2)].0.5CH(2)Cl(2), (I), 8-isopropoxy-7,7 bis(triphenylphosphine-P)-8,9:10,11-di-muH-7-platina-nido-undecaborane dichloromethane solvate, [Pt(C(3)H(18)B(10)O)(C(18)H(15)P)(2)].CH(2)Cl(2), (II), and 9-isopropoxy-7,7-bis(triphenylphosphine-P)-8,9:10,11-di-muH-7-platina-nido undecaborane dichloromethane solvate, [Pt(C(3)H(18)B(10)O)(C(18)H(15)P)(2)].CH(2)Cl(2), (III), has an 11-vertex nido polyhedral skeleton, with the 7-platinum centre ligating to two exo-polyhedral PPh(3) groups and an alkoxy-substituted polyhedral borane ligand. Compounds (II) and (III) are isomers. The Pt-B distances are in the range 2.214 (7)-2.303 (7) A for (I), 2.178 (16)-2.326 (16) A for (II) and 2.205 (6)-2.327 (6) A for (III). PMID- 11498606 TI - Eta5-(3)-1-methyl-1,2-dicarbollyl-eta5-2',5'-dimethylpyrrolylcobalt(III). AB - In the title compound, (eta(5)-2,5-dimethylpyrrolyl)[(7,8,9,10,11-eta)-7-methyl 7,8-dicarba-nido-undecaborato]cobalt(III), [3-Co[eta(5)-[2,5-(CH(3))(2) NC(4)H(2)]]-1-CH(3)-1,2-C(2)B(9)H(10)] or [Co(C(3)H(13)B(9))(C(6)H(8)N)], the Co(III) atom is sandwiched between the pentagonal faces of the pyrrolyl and dicarbollide ligands, resulting in a neutral molecule. The C-C distance in the dicarbollide cage is 1.649 (3) A. PMID- 11498607 TI - Tetrakis(dimethylamido)vanadium(IV). AB - The title compound, [V(C(2)H(6)N)(4)], (I), has non-crystallographic D(2d) molecular symmetry and contains an approximately tetrahedrally coordinated V atom with dimethylamido ligands. Each N atom features a nearly trigonal planar geometry. There are two independent molecules of (I) in the asymmetric unit. The results are compared with those previously reported for gas-phase electron diffraction studies [Haaland, Rypdal, Volden & Andersen (1992). J. Chem. Soc. Dalton Trans. pp. 891-895]. PMID- 11498608 TI - Comparative X-ray study of three nickel(II)-thiocyanate compounds. AB - Three cis nickel-dithiocyanate (SCN) complexes with different N,N'-bidentate bases have been prepared and their crystal structures determined: bis(2,2' bipyridine-N,N')bis(-ato-N)nickel(II), [Ni(SCN)(2)(C(10)H(8)N(2))(2)], bis(1,10 phenanthroline-N,N')bis(thiocyanato-N)nickel(II), [Ni(SCN)(2)(C(12)H(8)N(2))(2)], and bis(2,9-dimethyl-1,10-phenanthroline-N,N')bis(thiocyanato-N)nickel(II) monohydrate, [Ni(SCN)(2)(C(12)H(8)N(2))(2)].H(2)O. Distortions due to ligand size are discussed. PMID- 11498609 TI - A low-temperature neutron diffraction study of Mn12-acetate. AB - In the low-temperature region, where the dodecanuclear mixed-valence manganese carboxylate hexadecaacetatotetraaquadodecaoxododecamanganese bis(acetic acid) tetrahydrate, [Mn(12)O(12)(C(2)D(3)O(2))(16)(H(2)O)(4)].2C(2)HD(3)O(2).4H(2)O, displays unusual magnetic properties, its structure is similar to that previously determined at room temperature [Lis (1980). Acta Cryst. B36, 2042-2046], differing only by a small change in the configuration of one of the coordinated acetate groups, related to the formation of additional hydrogen bonds, and by the orientation of the methyl groups. Since most of the magnetization density of this system resides on the Mn atoms, the consequences of these rearrangements for the magnetic properties of the compound are small. PMID- 11498610 TI - A mixed-valence manganese(III)-manganese(IV) di-mu-oxo complex, [(cyclam)MnO]2(ClO4)2(NO3). AB - The title dinuclear di-mu-oxo-bis[(1,4,8,11-tetraazacyclotetradecane kappa(4)N)manganese(III,IV)] diperchlorate nitrate complex, [Mn(2)O(2)(C(10)H(24)N(4))(2)](ClO(4))(2)(NO(3)) or [(cyclam)MnO](2)(ClO(4))(2)(NO(3)), was self-assembled by the reaction of Mn(2+) with 1,4,8,11-tetraazacyclotetradecane in aqueous media. The structure of this compound consists of a centrosymmetric binuclear [(cyclam)MnO](3+) unit, two perchlorate anions and one nitrate anion. While the low-temperature electron paramagnetic resonance spectra show a typical 16-line signal for a di-mu-oxo Mn(III)/Mn(IV) dimer, the magnetic susceptibility studies also confirm a characteristic antiferromagnetic coupling between the electronic spins of the Mn(IV) and Mn(III) ions. PMID- 11498611 TI - Hydrogen-bonded adducts of ferrocene-1,1'-diylbis(diphenylmethanol): monomeric and dimeric 1:1 adducts with 1,2-bis(4-pyridyl)ethane and 1,2-diaminoethane. AB - In ferrocene-1,1'-diylbis(diphenylmethanol)-4,4'-ethylenedipyridine (1/1), [Fe(C(18)H(15)O)(2)].C(12)H(12)N(2), there is an intramolecular O-H...O hydrogen bond in the ferrocenediol component and a single O-H...N hydrogen bond linking the two components into a finite monomeric adduct. Ferrocene-1,1' diylbis(diphenylmethanol)-ethylenediamine (1/1), [Fe(C(18)H(15)O)(2)].C(2)H(8)N(2), crystallizes with Z' = 2 in space group P1, and there are two independent four-component aggregates in the structure, both of which are centrosymmetric. In the first type of aggregate, the molecular components are linked by O-H...N and N-H...O hydrogen bonds, in which both diamine N atoms participate; in the second type of aggregate, the diamine component is disordered over two sets of sites, but only one N atom is involved in the hydrogen bonding. PMID- 11498612 TI - [6-Amino-3-methyl-5-nitrosopyrimidine-2,4(1H,3H)-dionato]-sodium dihydrate at 150 K: coordination-polymer ladders linked by hydrogen bonds. AB - In the title compound, [Na(C(5)H(5)N(4)O(3))].2H(2)O, each Na cation is linked to three C(5)H(4)N(4)O(3)(-) anions and two water molecules, forming approximately octahedral NaNO(5) units; the cations and the anions form molecular ladders, and each ladder is linked to four neighbouring ladders by O-H...O and O-H...N hydrogen bonds. PMID- 11498613 TI - Sodium 2-nitroethanaloximate forms a layered-framework structure. AB - In the title compound, [Na(C(2)H(3)N(2)O(3))], the Na(+) cation lies on a centre of inversion in space group P2(1)/m and all the atoms of the anion lie on a mirror plane. Na is octahedrally coordinated by four O and two N atoms from six different anions and each anion is coordinated to six different Na(+) cations, forming chains of confacial octahedra which link the anion layers. Within these layers, the individual anions are linked by both O-H...O and C-H...O hydrogen bonds. PMID- 11498614 TI - Iodo(phthalocyaninato)chromium(III). AB - A new chromium(III)-phthalocyanine complex with the formula [Cr(C(32)H(16)N(8))I], or CrPcI where Pc is phthalocyanate(2-), has been obtained by the reaction of pure Cr powder with phthalonitrile under a stream of iodine vapour. The five-coordinate Cr atom is bonded to the four isoindole N atoms of the phthalocyaninate(2-) ligand and to one apical iodine ligand, and has a square pyramidal coordination geometry. The Cr(III) cation is significantly displaced [0.456 (2) A] from the N(4)-isoindole plane towards the I atom. The Cr-I bond is tilted 2.51 (4) degrees to the N(4)-isoindole plane. PMID- 11498615 TI - 5,6-Diphenyl[1,3]dithiolo[4,5-b]-dithiine-2-thione. AB - In the title compound, C(17)H(10)S(5), the dithiine ring adopts a boat conformation while the dithiole ring has an envelope conformation. The phenyl groups are planar and make dihedral angles of 40.7 (2) and 59.8 (2) degrees with the best plane of the thiine ring. The shortest intermolecular S...S contact is 3.305 (2) A. PMID- 11498616 TI - Investigations on benz[1,2]oxaphosphinane derivatives. AB - The structures of two compounds, 3-[1-(2-hydroxyethylamino)ethylidene]-2-methoxy 3,4-dihydro-1,2lambda(5)-benzoxaphosphinane-2,4-dione, C(13)H(16)NO(5)P, and 3-[1 (2-hydroxy-ethylamino)ethylidene]-2-methoxy-6-methyl-3,4-dihydro-1,2lambda(5) benzoxaphosphinane-2,4-dione, C(14)H(18)NO(5)P, have been studied and compared. The oxophosphinane rings have a half-chair conformation and extra six- and five membered rings are formed by intramolecular N-H...O hydrogen bonds. An intermolecular O-H...O hydrogen bond is also observed. PMID- 11498617 TI - Dimethyl N,N'-oxalamidodiethanoate. AB - The title compound, dimethyl 2,2'-(oxalyldiimino)diethanoate, C(8)H(12)N(2)O(6), exhibits a network of hydrogen bonds between amide and ester groups. Molecules lie on inversion centres and show a planar conformation for both the oxalamide and ester groups. The glycine residues adopt a conformation close to the polyglycine II structure. PMID- 11498618 TI - Diethyl 4-(2,5-dimethoxyphenyl)-2,6-dimethyl-1,4-dihydropyridine-3,5 dicarboxylate, conformational change with solvent of crystallization. AB - The title compound, C(21)H(27)NO(6), has been crystallized from ethanol containing nitrobenzene and shows the phenyl ring, B, in an ap conformation. This structure may be compared with that of the molecule crystallized from ethanol alone, in which the B ring is seen in an sp conformation. The isolation of this rotamer has implications for the understanding of the docking of calcium beta blocking dihydropyridine molecules with their receptor site. PMID- 11498619 TI - The lock-in phase in the urotropine-sebacic acid system. AB - The 1,10-decanedioic acid-1,3,5,7-tetraazatricyclo[3.3.1.1(3,7)]decane (1/1) system, C(10)H(18)O(4).C(6)H(12)N(4), was studied at 215 (2) K. Its analysis provides important information with regard to the long-standing acid-carboxylate controversy in the urotropine-alkanedioic acid system. In the present structure, all the chain end-groups display a clear acid character. The asymmetric unit of this commensurate modulated phase contains two molecules of diacid as well as two molecules of urotropine. Furthermore, the chain packing suggests a possible order parameter for the lock-in transition. PMID- 11498620 TI - Polymorphism in pentacene. AB - Pentacene, C(22)H(14), crystallizes in different morphologies characterized by their d(001)-spacings of 14.1, 14.5, 15.0 and 15.4 A. We have studied the crystal structure of the 14.1 and 14.5 A d-spacing morphologies grown by vapour transport and from solution. We find a close correspondence between the 14.1 A structure reported by Holmes, Kumaraswamy, Matzeger & Vollhardt [Chem. Eur. J. (1999), 5, 3399-3412] and the 14.5 A structure reported by Campbell, Monteath Robertson & Trotter [Acta Cryst. (1961), 14, 705-711]. Single crystals commonly adopt the 14.1 A d-spacing morphology with an inversion centre on both molecules in the unit cell. Thin films grown on SiO(2) substrates above 350 K preferentially adopt the 14.5 A d-spacing morphology, with a slightly smaller unit-cell volume. PMID- 11498621 TI - Triclinic and orthorhombic polymorphs of 2-iodo-4-nitroaniline: interplay of hydrogen bonds, nitro...I interactions and aromatic pi-pi-stacking interactions. AB - In the triclinic polymorph of 2-iodo-4-nitroaniline, C(6)H(5)IN(2)O(2), space group P-1, the molecules are linked by paired N-H...O hydrogen bonds into C(8)[R(2)(2)(6)] chains of rings. These chains are linked into sheets by nitro...I interactions, and the sheets are pairwise linked by aromatic pi-pi stacking interactions. In the orthorhombic polymorph, space group Pbca, the molecules are linked by single N-H...O hydrogen bonds into spiral C(8) chains; the chains are linked by nitro...O interactions into sheets, each of which is linked to its two immediate neighbours by aromatic pi-pi-stacking interactions, so producing a continuous three-dimensional structure. PMID- 11498622 TI - Hydrogen-bonded chains in N-(2-nitrophenyl)phenylamine. AB - Molecules of the title compound, C(12)H(10)N(2)O(2), are markedly non-planar. There is an intramolecular N-H...O hydrogen bond, and the molecules are linked into zigzag chains by a single C-H...O hydrogen bond. Comparisons are made with the supramolecular aggregation in isomeric amino-nitro derivatives, and in some N methylnitroanilines. PMID- 11498623 TI - N,N'-Bis[tris(hydroxymethyl)methyl]-ethanediamide: six O-H...O hydrogen bonds generate only a two-dimensional structure. AB - Molecules of the title compound, C(10)H(20)N(2)O(8), adopt a conformation which is almost centrosymmetric. The molecules are disordered over two sets of sites with an occupancy ratio of 0.94:0.06. In the major form, there are two intramolecular O-H...O hydrogen bonds [O...O 2.756 (4) and 2.765 (4) A; O-H...O 144 and 146 degrees ], in which the two amidic O atoms act as acceptors. In addition, there are four intermolecular O-H...O hydrogen bonds [O...O 2.650 (3) 2.666 (3) A; O-H...O 158-171 degrees ]; these link each molecule to six others in a continuous sheet structure which contains five distinct ring motifs, two of the S(7) type, two of the R(3)(3)(10) type and one of the R(2)(2)(22) type. PMID- 11498624 TI - N,N'-Dimethylpiperazinium(2+) phosphonoacetate: hydrogen-bonded anion sheets containing cation-templated R(6)(6)(28) rings. AB - In the title compound, C(6)H(16)N(2)(2+).2C(2)H(4)O(5)P(-), the cations lie across centres of inversion; in the anions, two of the H-atom sites have 0.50 occupancy. The anions are linked by short O-H...O hydrogen bonds [O...O 2.465 (3) 2.612 (3) A and O-H...O 165-171 degrees ] into sheets of alternating R(2)(2)(12) and R(6)(6)(28) rings, both of which are centrosymmetric; the cations lie at the centres of the larger rings linked to the anion sheet by N-H...O hydrogen bonds [N...O 2.642 (2) A and N-H...O 176 degrees ]. PMID- 11498625 TI - Hydrogen-bonded bilayers in piperazinium(2+) bis(mandelate) bis(methanol) solvate. AB - In the title compound, C(4)H(12)N(2)(2+).2C(8)H(7)O(3)(-).2CH(4)O, the cations lie across centres of inversion and are disordered over two orientations with equal occupancy; there are equal numbers of (R)- and (S)-mandelate anions present (mandelate is alpha-hydroxybenzeneacetate). The anions and the neutral water molecules are linked by O-H...O hydrogen bonds [O...O 2.658 (3) and 2.682 (3) A, and O-H...O 176 and 166 degrees] into deeply folded zigzag chains. Each orientation of the cation forms two symmetry-related two-centre N-H...O hydrogen bonds [N...O 2.588 (4) and 2.678 (4) A, and N-H...O 177 and 171 degrees] and two asymmetric, but planar, three-centre N-H...(O)(2) hydrogen bonds [N...O 2.686 (4) 3.137 (4) A and N-H...O 137-147 degrees], and by means of these the cations link the anion/water chains into bilayers. PMID- 11498626 TI - Hamigerin A and a hamigerin D decomposition product. AB - The sponge Hamigera tarangaensis has yielded eight new compounds and we report here the structure of one of these compounds, hamigeran A, C(20)H(25)BrO(5), or methyl 7-bromo-4beta,6-dihydroxy-1beta-isopropyl-3aalpha,8-dimethyl-5-oxo 1a,3a,4,5-tetrahydrocyclopenta[a]naphthalene-4-carboxylate, and the decomposition product of hamigeran D, C(21)H(28)BrNO(4), namely 2-(8-bromo-2beta,7-dimethyl-4 oxo-1,3alpha-benzoxazan-5-yl)-3-isopropylcyclopentylacetic acid. PMID- 11498627 TI - Differently coloured polymorphs of 4-[(2-nitrophenyl)azo]phenol. AB - The crystal structures of the brown-yellow and orange polymorphs of the title compound, 4-[(2-nitrophenyl)diazenyl]phenol, C(12)H(9)N(3)O(3), have been determined and their visible reflection spectra recorded. Both structures adopt a stacking arrangement with interstack hydrogen bonds. Ab initio and semi-empirical (AM1 and INDO-CISD) calculations were performed in order to rationalize the difference in colour. It can be attributed neither to the subtle distinctions in molecular geometry nor to the effect of intermolecular electrostatic interactions. The most probable origin of this difference is the mixing of intramolecular n --> pi* and intermolecular charge-transfer excitations. PMID- 11498628 TI - Zwitterionic 4-piperidinecarboxylic acid monohydrate. AB - The title compound, 4-piperidiniocarboxylate (isonipecotic acid), crystallizes as a zwitterion and incorporates one water molecule, i.e. C(6)H(11)NO(2).H(2)O. The piperidine ring adopts a chair conformation and the alpha-carboxylate group is oriented in the equatorial position. The combination of the interactions between the alpha-amino and alpha-carboxylate groups and the water molecules builds a three-dimensional assembly of hydrogen bonds. PMID- 11498629 TI - trans-1,2,3-Tris(1-hydroxycyclopropyl)cyclopropane. AB - The central three-membered ring in the title compound, trans-1,1',1"-cyclopropane 1,2,3-triyltris(cyclopropanol), C(12)H(18)O(3), shows pronounced asymmetry of the bond lengths, which is induced by the different orientations of the substituents. A network of hydrogen bonds links the molecules into sheets. PMID- 11498630 TI - 1,2-Bis(N-fluoro-p-toluenesulfonamido)ethane chloroform solvate. AB - The title compound, N,N'-difluoro-N,N'-ethylenedi-p-toluenesulfonamide, C(16)H(18)F(2)N(2)O(4)S(2).CHCl(3), is a novel stable compound of the N-F class of reagents containing two R(2)N-F functionalities. The compound, as the chloroform solvate, is the first such bis(N-F) compound to be structurally characterized. It adopts a solid-state structure in which the two aromatic rings are antiperiplanar and a combination of weak C-H...F and C-H...O hydrogen bonds [distances and angles range from 3.265 (4) to 3.439 (4) A and 150 to 170 degrees, respectively] and pi-stacking between the rings of different molecules (separations of 3.717 and 3.926 A) results in a solid-state structure containing well defined channels in which CHCl(3) solvent molecules are located. The N-F distances are 1.428 (3) and 1.433 (3) A. PMID- 11498631 TI - Dimethyl 2-[[2-(methoxycarbonyl)-1-(methoxycarbonylmethyl)pyrrol-4 yl]methylene]propanedioate. AB - The title compound, C(15)H(17)NO(8), is a pyrrole-ethene derivative with potential biological activity. Although a large part of the molecule is planar, there is no structural evidence for any conjugation push-pull effect across the ethylenic bond, which is usually observed for substituted ethylenes; pi-electron delocalization appears to be restricted to the 2-(methoxycarbonyl)pyrrole moiety. PMID- 11498632 TI - N-Z-Pro-D-Leu using synchrotron radiation data from a very small crystal. AB - The crystal structure of the neuroactive artificial dipeptide N benzyloxycarbonylprolyl-D-leucine, C(19)H(26)N(2)O(5), was solved using synchrotron radiation data collected on a very small crystal (20 x 20 x 380 microm). The molecules form hydrogen-bonded 2(1) helices. The acid carbonyl group does not participate in strong hydrogen bonds. This is interpreted as a consequence of close-packing requirements. PMID- 11498633 TI - Methoxy-ether and crown-ether derivatives of tetrahomodioxa- and octahomotetraoxacalix[4]arenes. AB - Three methoxy-ether and one methoxy-ether/crown-ether derivatives of p-tert butyltetrahomodioxa- and p-R-octahomotetraoxacalix[4]arenes (R = methyl, tert butyl, H) have been investigated. The first three compounds, 7,15,21,27-tetra tert-butyl-29,30,31,32-tetramethoxy-3,11 dioxapentacyclo[23.3.1.1(5,9).1(13,17).1(19,23)]ditriaconta 1(29),5,7,9(30),13,15,-17(31),19,21,23(32),25,27-dodecaene, C(50)H(68)O(6), 33,34,35,36-tetramethoxy-7,15,23,31-tetramethyl-3,11,19,27 tetraoxapentacyclo[27.3.1.1(5,9).1(13,17).1(21,25)]hexatriaconta 1(33),5,7,9(34),13,15,17(35),21,23,25(36),29,31-dodecaene, C(40)H(48)O(8), and 7,23-di-tert-butyl-33,34,35,36-tetramethoxy-3,11,19,27 tetraoxapentacyclo[27.3.1.1(5,9).1(13,17).1(21,25)]hexatriaconta 1(33),5,7,9(34),13,15,17(35),- 21,23,25(36),29,31-dodecaene, C(44)H(56)O(8), in the partial-cone or 1,2-alternate conformations, present the common feature of methoxy-ether self-inclusion, while the fourth, 42,43-dimethoxy-7,15,23,31 tetramethyl-3,11,19,27,34,37,40 heptaoxahexacyclo[15.15.9.1(5,9).1(21,25).0(13,41).0(29,33)]tritetraconta 5(42),6,8,13(41),14,16,21(43),22,24,29(33),30,32-dodecaene, C(42)H(50)O(9), adopts the 1,3-alternate conformation owing to the presence of a 1,3-polyether chain. PMID- 11498634 TI - Thermal isomerization pathway of 1-(4-nitrophenyl)-2-phenylimino-2,5-dihydro-1H pyrido[3,2-b]indole-3-carbonitrile discovered by laboratory powder data. AB - The evidence for thermal isomerization of the title compound, C(24)H(15)N(5)O(2), into 2-[(4-nitrophenyl)phenylamino]-5H-pyrido[3,2-b]indole-3-carbonitrile has been obtained as a consequence of crystal structure determinations from laboratory powder data. PMID- 11498635 TI - 2-(N,N-Diphenylamino)benzoic acid. AB - Shaped like a distorted propeller, molecules of the title compound, C(19)H(15)NO(2), form centrosymmetric dimers in the crystalline phase in which the carboxy groups are linked through two hydrogen bonds. These dimers are arranged in columns held together via dispersive interactions between the phenyl moieties. The N atom and the three surrounding C atoms lie almost in the same plane, which implies that the lone electron pair of the N atom is involved in conjugation with the pi systems of the phenyl fragments. PMID- 11498636 TI - 1H-Pyrrole-2-carbothioamide. AB - The reaction of ethoxycarbonylpyrrole-2-thiocarboxamide with sodium hydroxide affords the known title compound, C(5)H(6)N(2)S, whose structure shows a three dimensional hydrogen-bonded lattice. PMID- 11498637 TI - N-tert-Butyl-N'-(5,7-dimethyl-1,8-naphthyridin-2-yl)urea. AB - The title compound, C(15)H(20)N(4)O, has been synthesized as an AADD recognition unit for quadruple hydrogen bonds. All non-H atoms of the molecule apart from two methyl groups of the tert-butyl group lie in a common plane. An intramolecular hydrogen bond is formed connecting two N atoms. In the solid state, the title compound crystallizes as a centrosymmetric dimer connected by N-H...O=C interactions with an N...O distance of 2.824 (2) A. PMID- 11498638 TI - MEM(TCNQ)2 at room temperature and 10 K, and the absence of a spin-Peierls transition. AB - Precise X-ray determinations of the crystal structure of the 1:2 complex of N ethyl-N-methylmorpholinium and 7,7,8,8-tetracyano-p-quinodimethanide, abbreviated as MEM-TCNQ or MEM(TCNQ)(2) (C(7)H(16)NO(+).2C(12)H(4)N(4)(0.5-)), have been performed at 293 and at 10 K. Evidence for the expected spin-Peierls transition at 19 K is not found, and this may follow from radiation damage to the crystal or from insufficient equipment sensitivity. PMID- 11498639 TI - Diisopropylammonium hydrogen difluoride. AB - By the reaction of diisopropylamine with concentrated hydrofluoric acid, the title compound, C(6)H(16)N(+).HF(2)(-), was synthesized. The cations and anions are connected via hydrogen bonds to form chains along [100]. The interlocked polymeric zigzag chains are hexagonally surrounded by six neighbouring chains. The F(-) anion is strongly hydrogen bonded to HF [F-H 0.98 (4), H...F 1.34 (4), F...F 2.3125 (16) A and F-H...F 174 (3) degrees]; a better description is that of a non-centred hydrogen difluoride anion. PMID- 11498640 TI - Conformational polymorphs of 22-cyano-N-methyl-5-phenylpent-2-en-4-ynamide. AB - Although the two polymorphic modifications, (I) and (II), of the title compound, C(13)H(10)N(2)O, crystallize in the same space group (P2(1)/c), their asymmetric units have Z' values of 1 and 2, respectively. These are conformational polymorphs, since the molecules in phases (I) and (II) adopt different rotations of the phenyl ring with respect the central 2-cyanocarboxyaminoprop-2-enyl fragment. Calculations of crystal packing using Cerius(2) [Molecular Simulations (1999). 9685 Scranton Road, San Diego, CA 92121, USA] have shown that (I) is more stable than (II), by 1.3 kcal mol(-1) for the crystallographically determined structures and by 1.56 kcal mol(-1) for the optimized structures (1 kcal mol(-1) = 4.184 kJ mol(-1)). This difference is mainly attributed to the different strengths of the hydrogen bonding in the two forms. PMID- 11498642 TI - Atrioventricular filling dynamics, diastolic function and dysfunction. AB - Left ventricular diastolic dysfunction is associated with slowing of LV relaxation and a decrease in LV chamber compliance. This impairment of function leads to changes in filling velocities as measured by pulsed wave Doppler echocardiography in the pulmonary veins and across the mitral valve, and in intraventricular flow propagation velocity as measured by color M-mode Doppler. This paper explores some of the physiology of LV filling in a clinical context. PMID- 11498643 TI - Diastolic dysfunction as a cause of exercise intolerance. AB - Tachycardia accompanying exercise shortens the duration of diastole, reducing the time available for the left ventricular (LV) filling. Thus, the LV must fill more rapidly for the stroke volume to increase (or even be maintained) during exercise. Normally, this is accomplished without requiring an excessive increase in left atrial (LA) pressure by an acceleration of LV relaxation and a fall in LV early diastolic pressure during exercise. This response is lost following the development of heart failure due to systolic dysfunction, both in experimental animals and in patients. In fact, in such situations, LV relaxation slows and LV early diastolic pressure increases due to exercise. Thus, any diastolic dysfunction present at rest in CHF during systolic dysfunction is exacerbated during exercise. Similarly, patients with primary diastolic dysfunction heart failure with preserved systolic function may not be able to augment LV filling rates without an abnormal increase in LA pressure. Thus, diastolic dysfunction may contribute to exercise intolerance, both in systolic dysfunction and primary diastolic dysfunction. Acute studies suggest that treatment with angiotensin II receptor blockers or verapamil may improve exercise tolerance in some patients with primary diastolic dysfunction. PMID- 11498644 TI - Diastolic ventricular interaction and ventricular diastolic filling. AB - Because the ventricles share a common septum, the filling of one may influence the compliance of the other, a phenomenon known as direct diastolic ventricular interaction (DVI). This interaction is markedly enhanced when the force exerted by the surrounding pericardium is raised (pericardial constraint). In health, in the resting state, we operate near the top of the flat component of a J-shaped pericardial stress-strain relation. Therefore, pericardial constraint (and hence DVI) is only minor. When right ventricular volume/pressure acutely increases, such as during exercise, massive pulmonary embolism, or right ventricular infarction, pericardial constraint increases and significant DVI develops. In this setting, the measured left ventricular intracavitary diastolic pressure markedly overestimates the true left ventricular filling pressure, because the external forces must be subtracted. Although the pericardium can grow during chronic cardiac enlargement, we present evidence that in certain chronic disease processes, including heart failure, DVI may also be important. PMID- 11498646 TI - LV hypertrophy and diastolic heart failure. PMID- 11498645 TI - Modulation of left atrial function by ventricular filling impairment. AB - Left atrial function is an important determinant of ventricular filling. Assessment of the complex role that the atrial cavity exerts in the ventricular filling process can be made noninvasively. Computing the net instantaneous difference between mitral and pulmonary venous flow is an approach which permits the construction of the left atrial volume curve throughout the cardiac cycle (as well as the left ventricular volume curve during diastole), and to quantify the 3 different functions that the cavity performs. In particular, increasing degrees of ventricular filling impairment are met by mechanical left atrial adaptations which basically rely on the Starling mechanism, with the reservoir/pump complex activated to the limit of the preload reserve of the cavity. At end-stage left ventricular dysfunction, however, the atrial reservoir and the booster pump function decline and conduit takes precedence, suggesting afterload mismatch, impaired atrial compliance and, perhaps, depressed atrial contractility. Increased wall stiffening and reduced elastic recoil induced by chronic atrial distension might explain the additional power of atrial size in stratifying prognostically patients with left ventricular dysfunction. PMID- 11498647 TI - Beneficial effects of nitric oxide on cardiac diastolic function: 'the flip side of the coin'. AB - Modulation by NO of systolic myocardial function received widespread attention but most studies focused on potential negative inotropic properties of NO. The very original observations on the effects of NO on myocardial contraction already provided evidence that NO modified myocardial contractile performance mainly through a relaxation-hastening effect (i.e. earlier onset of relaxation) and through an increase in myocardial distensibility. The present review discusses the relaxation hastening and distensibility-increasing effects of NO in experimental preparations, in the normal human heart, in left ventricular hypertrophy of aortic stenosis, in the human allograft and in dilated nonischemic cardiomyopathy. This 'diastolic flip side' of the myocardial effects of NO appears to be beneficial especially for patients who are dependent on the LV Frank-Starling response to maintain cardiac output. PMID- 11498648 TI - Load dependent diastolic dysfunction in heart failure. AB - Congestive heart failure may result from cardiovascular overload, from systolic or from diastolic dysfunction. Diastolic left ventricular dysfunction may result from structural resistance to filling such as induced by pericardial constraint, right ventricular compression, increased chamber stiffness (hypertrophy) and increased myocardial stiffness (fibrosis). A distinct and functional etiology of diastolic dysfunction is slow and incomplete myocardial relaxation. Relaxation may be slowed by pathological processes such as hypertrophy, ischemia and by asynchronous left ventricular function. The present contribution analyses the occurrence of slow and incomplete myocardial relaxation in response to changes in systolic pressure and in response to changes in venous return. The regulation of myocardial relaxation by load is critically dependent on the transition from myocardial contraction to relaxation, which occurs in dogs when 82% of peak isovolumetric pressure has developed or at a relative load of 0.82. This corresponds to early ejection in normal hearts, but is situated even before aortic valve opening in severely diseased hearts. When load is developed beyond this transition, relaxation becomes slow and even incomplete. This is load dependent diastolic dysfunction. Load dependent diastolic dysfunction occurs in normal hearts facing heavy afterload and in severely diseased hearts even with normal hemodynamic parameters. This dysfunction should contribute to elevating filling pressures in most patients with severe congestive heart failure. This dysfunction can be reverted by decreasing systolic pressures or by decreasing venous return. Load dependent diastolic dysfunction gives us an additional reason to aggressively treat CHF patients with diuretics and vasodilators. PMID- 11498649 TI - Mechanisms and efficacy of LV pre-excitation for patients with heart failure and supra-normal systolic function. AB - Ventricular hypertrophy due to genetic mutations of sarcomeric proteins or that associated with long-standing hypertension typically yields a cavity with hyperdynamic ejection, elevated diastolic pressures, and limited filling volumes. The net result is reduced reserve capacity, dyspnea with exertional intolerance, and chest discomfort despite normal appearing coronary vessels. In addition to pharmacologic therapy by agents having negative inotropic effects, recent studies have examined the potential of ventricular pacing using right apical pre excitation as a treatment for these disorders. This form of pacing can increase end-systolic volume and reduce cavity obliteration in both forms of the disease, yet has no demonstrable acute benefit on diastolic function. Chronic therapy trials have yielded mixed results, with more favorable responses observed in older patients particularly those with hypertensive hypertrophic disease. These data have also highlighted the importance of enhancing systolic reserve rather than diastolic function as a key therapeutic effect from pacing therapy. This review discusses the mechanisms by which pacing with ventricular pre-excitation acutely influences ventricular function, and summarizes results of recent clinical trials, putting the data into perspective regarding the relative role of systolic versus diastolic effects in these patients. PMID- 11498650 TI - Systolic hypertension; alpha blockers and prostatism; are beta blockers still indicated; diabetes, obesity, and hypertension -- comments on the JCH contents. PMID- 11498651 TI - The challenge of controlling systolic blood pressure: data from the National Health and Nutrition Examination Survey (NHANES III), 1988--1994. AB - Clinicians tend to focus on diastolic blood pressure (DBP), rather than systolic blood pressure (SBP), to identify and treat hypertension. The authors used data from the National Health and Nutrition Examination Survey (NHANES III, 1988- 1994) Mobile Examination Center to examine the distributions of SBP and DBP in treated and untreated individuals with hypertension. We identified the percentage of the hypertensive population with SBP controlled to less than 140 mm Hg and the percentage with DBP controlled to less than 90 mm Hg, stratified by treatment status, gender, race, and ethnicity. Individuals were classified as having hypertension if they had SBP of more than 140 mm Hg or DBP of more than 90 mm Hg, or if they were taking medication for hypertension. A weighted analysis was performed to project the results to the entire U.S. population from 1988--1994; these totals were further estimated for the year 2000 by extrapolation. For men, women, whites, African Americans, and Hispanics, SBP control rates were uniformly poorer than DBP control rates. The difference persisted when subgroups were categorized according to treatment status. The disparity in SBP and DBP control rates was especially great for women: only 50% of treated white women with hypertension had SBP control, but 92% had DBP control. The prevalence of isolated systolic hypertension was greater than 50% among all individuals with hypertension in the 55--60-year age group and increased with age thereafter. A greater emphasis on SBP is needed to improve population blood pressure control. (c)2001 Le Jacq Communications, Inc. PMID- 11498652 TI - Doxazosin added to single-drug therapy in hypertensive patients with benign prostatic hypertrophy. AB - The purpose of this study was to evaluate the efficacy and safety of the addition of doxazosin in the treatment of hypertensive patients who are being treated on another antihypertensive drug. The open-labeled, noncomparative, multicenter study was carried out in 2363 male hypertensive outpatients > 40 years of age, under reasonable control with single antihypertensive drug treatment (diastolic blood pressure < 95 mm Hg), and diagnosed with benign prostatic hypertrophy. Doxazosin was started at a dose of 1 mg/day, which was increased at 2-week intervals to 2 mg/day and 4 mg/day. The study lasted 14 weeks. Blood pressure and heart rate were measured at each of the visits. At baseline and after 14 weeks of treatment, prostatism symptoms were quantified with the International Prostate Symptom Score and quality of life was determined with the American Urology Association Committee Guidelines. Adverse effects were recorded. At the fourth visit, when the patients were taking 4 mg of doxazosin, the blood pressure reduction was 10.7 +/- 3/7.1 +/- 7.1 mm Hg. The decrease in diastolic blood pressure was significantly more marked in patients treated with beta blockers than in patients on calcium antagonists or angiotensin-converting enzyme inhibitors. For systolic blood pressure, decreases were larger in patients treated with diuretics than with calcium antagonists or angiotensin-converting enzyme inhibitors. Prostatism symptoms decreased from 15 +/- 5.8 points to 7.9 +/ 4.3 points (p is less than 0.001) and quality of life improved. Tolerability was good, with only a 4.4% cumulative incidence of adverse effects related to doxazosin. The patients who experienced adverse effects were older and their final blood pressures were lower. The results of this open-label study suggest that the addition of doxazosin to another antihypertensive drug in hypertensive patients with benign prostatic hypertrophy is well tolerated and leads to a reduction in prostatic symptoms. The additional beneficial effects on blood pressure suggest that the use of doxazosin may provide a rational approach to this category of patients.(c)2001 Le Jacq Communications, Inc. PMID- 11498653 TI - Management of high-risk hypertensive patients with diabetes: potential role of angiotensin II receptor antagonists. AB - Uncontrolled hypertension leads to an increased risk of cardiovascular disease and stroke. Hypertensive patients with concomitant type 2 diabetes are at even greater risk of cardiovascular complications; also, this high-risk patient population is at increased risk of renal disease and, ultimately, renal failure. Prospective morbidity and mortality trials have demonstrated that tight blood pressure control improves the cardiovascular prognosis and provides target organ protection. Current treatment guidelines recommend a target blood pressure of < 130/85 mm Hg for patients with hypertension and diabetes. Angiotensin II (A-II), a major component of the renin-angiotensin system, plays an essential role in the pathophysiology of hypertension and diabetes-related renal disease. Currently, the treatment of choice for hypertensive patients with diabetes is angiotensin converting enzyme (ACE) inhibition, but most of the data are limited to patients with type 1 diabetes. Although ACE inhibition is clearly a mechanism for blocking A-II formation, inhibition at this site may not be complete, as alternate pathways exist for A-II formation. Thus, for interrupting the renin-angiotensin system, A-II receptor antagonists theoretically provide advantages over ACE inhibitors in that they directly inhibit A-II by binding to the AT(1) receptor subtype. The objectives of this review are to: 1) provide an overview of the associated risk of cardiovascular complications with concomitant hypertension and diabetes; 2) demonstrate the cardiovascular benefits of effective blood pressure control in this patient population; 3) review the current treatment guidelines for managing high-risk hypertensive patients; and 4) discuss major, ongoing clinical studies with A-II receptor antagonists in patients with concomitant hypertension, type 2 diabetes, and renal disease. (c)2001 Le Jacq Communications, Inc. PMID- 11498654 TI - The beta blockers: are they as protective in hypertension as in other cardiovascular conditions? AB - Beta blockers are frequently used to treat hypertension because of their well established safety and efficacy. Large clinical trials yield a 12%--20% decline in cardiovascular end points in hypertensive patients treated with beta blockers. However, beta blockers account for only 11% of antihypertensive prescriptions, and their use appears to be declining as newer agents with fewer side effects become available. The metabolic side effects of beta blockers have recently been examined. While they may raise triglycerides, lower high-density lipoprotein cholesterol, induce glucose intolerance, and possibly unmask diabetes, these effects have not been shown to impact their clinical effectiveness. For hypertension, beta blockers are still recommended as first-line therapy in many patients, particularly those at high risk for cardiovascular disease. They are also indicated for other cardiovascular disorders, such as congestive heart failure and postmyocardial infarction, in which mortality reductions exceed that seen with hypertension treatment in patients without cardiovascular complications. (c)2001 Le Jacq Communications, Inc. PMID- 11498655 TI - The African American Study of Kidney Disease and Hypertension (AASK): new findings. AB - In September, 2000, the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health called an early halt to the amlodipine arm of the African American Study of Kidney Disease and Hypertension (AASK) trial after careful deliberation by an independent data and safety monitoring board. An interim analysis of the AASK at 3 years revealed a renoprotective effect of the angiotensin-converting enzyme inhibitor ramipril as compared to the dihydropyridine calcium channel blocker (DHP-CCB) amlodipine in patients with mild to moderate renal insufficiency. This differential effect was independent of the blood pressure (BP) levels reached and was evident in proteinuric patients and suggestive in patients with baseline proteinuria < 300 mg/d, but was not conclusive. The AASK trial data suggest that DHP-CCBs should be used cautiously in the presence of mild to moderate renal insufficiency. Judgment should be reserved for the use of other CCBs, such as verapamil or diltiazem, since these are fundamentally different CCBs with the potential for a different impact on hypertensive nephrosclerosis. The blinded observation period for AASK will be completed at the end of September, 2001, at which time additional, clinically useful information is expected to become available. (c)2001 Le Jacq Communications, Inc. PMID- 11498657 TI - New stuff about the diabetic kidney. PMID- 11498656 TI - Obesity and hypertension: a growing problem. PMID- 11498658 TI - Renal insufficiency predicts cardiovascular disease in high-risk individuals: the benefit of ramipril in the HOPE study. Heart Outcomes and Prevention Evaluation. PMID- 11498659 TI - Micardis/HCT (telmisartan/hydrochlorothiazide). PMID- 11498660 TI - Angioneurotic edema. AB - Angioneurotic edema is nonpitting edema involving the face, lips, tongue, larynx, and other locations. It may occur with use of angiotensin- converting enzyme inhibitors, vasopeptidase inhibitors, and, less frequently, angiotensin receptor blockers. Most episodes occur within the first 6 months of therapy. Predisposing factors for converting enzyme inhibitor angioedema include previous idiopathic angioedema, African American race, and transplant-related immunocompromise. Treatment is directed to preventing airway compromise. (c)2001 Le Jacq Communications, Inc. PMID- 11498675 TI - Molecular and cellular mechanisms of allergic disease. AB - The molecular and cellular mechanisms mediating the allergic inflammatory cascade involve multiple mediators, cell types, and pathways. Of particular interest are the pathways regulated by the T(H)2 lymphocyte, which result in release of IL-4 (important to IgE synthesis) and IL-5 (important to eosinophil proliferation). IL 4 regulates differentiation of naive T(H)0 cells to develop a T(H)2 phenotype and stimulates B cells to produce IgE. Cross-linking by allergen of IgE affixed to high-affinity receptors on mast cells and basophils triggers degranulation and the release of preformed inflammatory mediators (important to the early phase response), and subsequently initiates synthesis and the release of lipid mediators and cytokines (which may contribute to the late phase response). Eosinophils may also play a prominent role in the development of bronchial hyperreactivity. IL-5, which is a lineage-specific eosinophil growth factor, increases the formation of eosinophils from progenitor cells and, in concert with CCR3 active chemokines, increases their trafficking to sites of allergic inflammation. An improved understanding of the basic mechanisms of allergic inflammation has led to the discovery of molecular targets involved in the initial events of the inflammatory cascade. Potential targets for the development of novel therapies for allergic disease include IgE, the T(H)2 lymphocyte, and T(H)2-derived cytokines, IL-4 and IL-5. PMID- 11498676 TI - Cytokine-directed therapies for asthma. AB - Increasing knowledge of the pathophysiologic roles of various cytokines in atopic diseases has provided the basis for the development of novel therapies. Strategic approaches for cytokine inhibition include the blocking of transcription factors that lead to their expression, blockade after their release, cytokine receptor antagonism, and the inhibition of signaling pathways that are activated after cytokine-receptor binding. The proinflammatory cytokines IL-5, IL-4, IL-13, and TNF-alpha are among the therapeutic targets. Results with a humanized anti-IL-5 have been disappointing. Although successful in markedly reducing circulating eosinophils and in preventing eosinophil accumulation in airways, the humanized anti-IL-5 was unable to affect early or late responses to allergen or to reduce airway reactivity to methacholine challenge in patients with asthma. On the other hand, a soluble IL-4 receptor antagonist has shown clinical benefits for patients with moderate asthma who require daily inhaled corticosteroids. Agents that target IL-13 and TNF-alpha remain to be evaluated in asthmatic inflammation. The use of cytokines with anti-inflammatory effects may also have therapeutic value. The evaluation of such agents in human beings, including IL-10, IL-12, and IFN gamma, is at a preliminary stage, but so far results have not been encouraging. PMID- 11498677 TI - Experiences with monoclonal antibody therapy for allergic asthma. AB - Identification of the central role IgE plays in the pathogenesis of allergic diseases made it a key target for therapy. The first selective anti-IgE therapy, a unique humanized monoclonal anti-IgE antibody (omalizumab), binds with high affinity to the Fc(epsilon)RI receptor binding site on IgE, thereby reducing the amount of free IgE available to bind to Fc(epsilon)RI receptors on mast calls, basophils, and other cells. In addition, administration of omalizumab indirectly reduces Fc(epsilon)RI receptor density on cells involved in allergic responses. In two bronchoprovocation trials involving patients with mild allergic asthma, omalizumab attenuated both early- and late-phase allergic responses. Omalizumab was subsequently evaluated as a treatment for asthma in large, multicenter, randomized, double-blind phase II and III trials involving patients with moderate to severe asthma who required corticosteroid therapy. When added to treatment with oral or inhaled corticosteroids, omalizumab reduced symptoms and exacerbations, improved lung function and quality of life, and reduced the need for rescue medications. These benefits persisted even in the "corticosteroid reduction" phase of these trials, when omalizumab treatment was shown to allow patients to reduce or discontinue their inhaled and/or oral corticosteroids. These effects of omalizu-mab in improving asthma control, as well as its excellent safety profile, may ultimately make this agent a useful addition to the physician's armamentarium of treatments for asthma. PMID- 11498678 TI - Experience with monoclonal antibodies in allergic mediated disease: seasonal allergic rhinitis. AB - Current therapies for the treatment of seasonal allergic rhinitis include allergen avoidance; pharmacologic interventions such as sympathomimetics, topical and systemic cortico-steroids, and chromones; and immunotherapy. In an attempt to create a novel therapy, therapeutic agents have been designed to inhibit IgE responses that are intimately involved in the induction of the allergic response. Omalizumab, a humanized monoclonal antibody against IgE, represents a novel therapeutic intervention for seasonal allergic rhinitis. Complex formation of omalizumab with serum-free IgE reduces the amount of IgE available for binding to effector cells and thus has the potential to reduce IgE-mediated allergic symptoms. Clinical trial results confirmed that omalizumab reduces free IgE to a level that is associated with suppressed allergic symptoms, reduces concomitant rescue medication use, and improves rhinitis-specific quality of life. Patients treated with omalizumab during one pollen season can be re-treated during the subsequent season with minimal risk of adverse events. Omalizumab is non-allergen specific and does not induce acute anaphylaxis because of the lack of IgE crosslinking with basophil- or mast-cell-bound IgE. Furthermore, subcutaneous or intravenous administration of omalizumab does not invoke the generation of anti omalizumab antibodies. Thus, omalizumab represents a novel agent that should assist in the treatment of allergic rhinitis. PMID- 11498679 TI - [The immortalized cell lines induced by human papillomavirus type 16 (HPVS 16) YY1 binding-site mutants have some characteristics of transformed cells]. AB - OBJECTIVE: To study the biological characteristics of the immortalized cell lines of human keratinocytes from foreskin induced by human papillomavirus type 16 (HPV 16) YY1 binding-site mutants. METHODS: The cellular extracts of immortalized cell lines were prepared and the cellular endogenous p53 proteins were determined with Western blot. The cellular telomerase activities were analyzed with TRAP. The immortalized cells were incubated in a soft agarose medium containing 10% FCS and the anchorage-independent growth abilities of the tested cell lines were evaluated. RESULTS: Western blot showed that the endogenous p53 protein in all 4 tested clones were undetectable. The telomerases were clearly identified within the 4 tested cell lines, and showed an increasing activity along with the passages of the cells. All the clones were unable to grow in the soft-agarose medium during the earlier passages, but three of them showed anchorage independent-growth at the passages of 30-35. CONCLUSIONS: The immortalized cell lines induced by the HPV 16 YY1 binding-site mutants have active telomerase activity and ability of anchorage-independent growth. PMID- 11498680 TI - [A strain of influenza A H9N2 virus repeatedly isolated from human population in China]. AB - OBJECTIVE: To understand the subtype of surface proteins (hemagglutinin and neuraminidase) and antigenic character of the isolate of influenza virus, as well as to know its origin. METHODS: The virus isolation was performed on MDCK cells. The identification and antigenic analysis of the isolate were carried out with HI and NI tests. The serum antibody in population against the isolate was detected with HI and neutralization assays. The individual investigation of the patient from whom the H9N2 virus was isolated was followed. RESULTS: The isolate was H9N2 subtype of influenza A virus. It was G9 like strain. Its antigenicity was different from that of H9N2 viruses which were previously found in humans, chickens and pigeons. The HI and neutralization titers to H9N2 virus in convalescent serum of the patient reached 1:400 and > or =1: 640, respectively. The HI antibody titer 1:25 to H9N2 virus was also detected in the serum of patient's mother. CONCLUSIONS: The isolate is H9N2 subtype of influenza A virus and belongs to G9 like strain. The antigenic character of the isolate was different from that of H9N2 viruses found previously in humans, chickens and pigeons. The greatest possibility was that her mother had contacted with birds, especially chickens carrying H9N2 virus, then transmitted to her or she breathed in the air borne with H9N2 virus particles directly. PMID- 11498681 TI - [Cloning and primary analysis of 3 'end genome of two alphaviruses isolated from Hainan Province of China]. AB - OBJECTIVE: To further characterize the HBbl7 and Ml viruses isolated from Hainan Province by molecular biology. METHODS: The fragment containing partial El and 3'untranslated region of each of the viral isolate was amplified by RT-PCR, then subcloned. Recombinant was screened and sequenced. Analysis and comparison were carried out. RESULTS: 1.6kb fragment was amplified from HBbl7 and a 1.3kb was from Ml Comparison of sequences showed, in 3'untranslated region the nucleotide homology between HBbl7 and T48, the prototype of Ross River virus, is 99%, between Ml and Sagiyama virus is 98%. In the partial El sequence, the nucleotide (amino acid) homology between HBbl7 and T48 is 99%(99%),between Ml and Sagiyama virus is 97%(99%), and between Ml and Cetah virus is 94% (98%). CONCLUSIONS: Sequence analysis showed HBb17 virus belongs to Ross River virus, Ml virus belongs to Sagiyama virus or Cetah virus. PMID- 11498682 TI - [Effect of Sindbis XJ-160 virus structural proteins on BHK-21 cell]. AB - OBJECTIVE: To study the effect of Sindbis XJ-160 virus structural proteins on BHK 21 cell. METHODS: We constructed the expressing vector pcDNA3.1 ABC that contains the XJ-160 virus structural protein gene and then transfected it to BHK-21 cell. RESULTS: The expressed products can be detected by RNA dot blot hybridization or immUnofluorescent assay. The cytopathic effect was observed in BHK-21 cells transfected with pcDNA3.1 ABC. There was also condensed chromatin found accumulated. BHK-21 cell death and marked cell cycle changes were observed. CONCLUSIONS: These results indicate that cell apoptosis might be one of the mechanisms inducing cell death by XJ - 160 virus and its structural protein gene probably plays a role in this programmed cell death. PMID- 11498683 TI - [Cell cycle G2 arrest, cell death and nuclear localization induced by human immunodeficiency virus type 1 protein R (VPR) in cervical cancer cells]. AB - OBJECTIVE: To explore the possibility of using Vpr as a biological agent for anticancer therapy. METHODS: A HIV-1 wild type Vpr and a mutant VprX gene were expressed and tested in a cervical cancer HeLa cell line. The ability of Vpr to induce cell cycle G2 arrest and cell death was measured by flow cytometric analysis. Nuclear localization of Vpr was visualized by fluorescent light emitted from the green fluorescent protein (GFP) -Vpr fusion protein. RESULTS: Consistent with early reports, Vpr, not VprX, induced G2 arrest. However, both Vpr and VprX were able to induce cell death in HeLa cells. We also demonstrated that the nuclear localization of Vpr could be determined by using a GFP-Vpr fusion protein. CONCLUSIONS: This is the first report documenting in mammalian cells that G2 arrest and cell death induced by Vpr are two independent functions. The unique biological properties of Vpr shown in cervical cancer cells suggest that Vpr may be a useful biological agent for anti -cancer therapy. PMID- 11498684 TI - [Study of hepatitis C virus specific immune responses in anti-HCV positive patients without hepatitis C viremia]. AB - OBJECTIVE: To study the hepatitis C virus specific immune responses in anti - HCV positive patients without hepatitis C viremia. METHODS: 15 anti-HCV positive patients without hepatitis C viremia, 15 patients with chronic HCV infection and 15 normal controls were selected for this study. The T cell responses, NK cell (natural killer cells) activity, cytokine production and HCV specific antibodies were detected by MTT, LDH release and ELISA. RESULTS: Our study showed that the T cell proliferative reaction of patients without hepatitis C viremia was significantly higher than that of patients with chronic HCV infection and normal controls and the T cell response for HCV core antigen were higher than NS3 and N54, but there was no significant proliferative response to NS5 antigen. We also found that there were no differences in anti-HCV antibody production and NK cell activity between the two groups and the level of IFN-gamma in patients without hepatitis C viremia was higher than that in patients with persistent HCV infection. CONCLUSIONS: There are a lot of advantageous changes of HCV specific humoral and cellular immune response in anti-HCV positive patients without hepatitis C viremia, these immune responses may play a role in clearance of HCV. PMID- 11498685 TI - [Molecular epidemiological study on HIV-1 infection in Shandong Province]. AB - OBJECTIVE: The aim of this study was to understand the distribution of HIV-1 strain subtypes and their variation in Shandong Province and to analyze their source and predict their epidemic trend. METHODS: We collected the whole blood from 25 anti-HIV-1 antibody positive cases and separated the peripheral blood mononuclear cells. The provirus DNA was extracted and amplified by nested-PCR, after which the HIV-1 membrane protein gene (env) C2-V3 region was sequenced and its subtype was analyzed. RESULTS: Among 25 HIV-1 antibody positive cases, 24 HIV 1 env gene fragments were amplified and analyzed.5 subtypes of Group HIV-1 M were identified among these cases,that are subtypes A, B, B', C and E. 13 occupational blood donors were all classified as subtype B' (Thailand B'). A, B, C or E subtypes were discovered in 10 returned laborers working abroad and 1 female spouse of one laborer. CONCLUSIONS: The condition of HIV-1 subtype infection was complicated in Shandong Province. So, it is important to strengthen the surveillance and control of HIV-1 subtype strain infection in high risk populations especially the occupational blood donors and returned laborers working abroad to prevent its spread in Shandong Province. PMID- 11498686 TI - [Detection of anti-HIV-1/2 and HIV nucleic acid in population of non-remunerated blood donors in Shenzhen]. AB - OBJECTIVE: To study the prevalent status of HIV-1 in population of non remunerated blood donors in Shenzhen. METHODS: 46,095 non-remunerated blood donors were tested for anti-HIV-1/2 by ELISA. The donors of anti-HIV positive were further detected for HIV DNA from peripheral blood mononuclear cells (PMBCs) by nested-PCR and HIV RNA from plasma by RT-PCR. The partial genome of env of 2 blood donors were sequenced. RESULTS: The anti-HIV-1 was tested positive in 7 of 46 095 voluntary blood donors and the positive rate was 0.015%. In these 7 non remunerated blood donors of anti-HIV-1 positive,7 were positive for HIV DNA in PMBCs and 5 positive for HIV RNA in plasma. The sequence analysis showed that 2 donors were infected by HIV-1 subtype E strains. CONCLUSIONS: There exists HIV-1 subtype E infection in population of non-remunerated blood donors in Shenzhen. It is essential to detect and monitor strictly the population of non-remunerated blood donors. PMID- 11498687 TI - [Experimental infection of a novel nonenveloped DNA hepatitis virus in Rhesus monkey]. AB - OBJECTIVE: To test the susceptibility of the Rhesus monkey to the TT virus and to establish its transmission route. METHODS: Rhesus monkeys were administered orally and intravenously with the inoculum that was prepared with feces collected from a patient at the incubation period. Passage study was done with monkey' s positive feces. The blood, bile and feces were tested with polymerase chain reaction (PCR), and the liver, the jejunum tissues with in situ hybridization. RESULTS: The viremia occurred 4-7 days after intravenous inoculation and 7-10 days after oral administration. The virus was also excreted in the feces in a few days after oral infection and simultaneously with viremia after intravenous inoculation. The virus was also detected in the bile during the viremic phase. There was a prolonged carrier state that the viremia and fecal virus excretion persisted for more than 6 months. No serum transaminase elevation was found during the infection. There were virus signals in hepatocytes in columnar epithelium and lamina propria cells of jejunum villi, but no significant pathology was demonstrated in both sites. CONCLUSIONS: The liver infection of Rhesus monkey was established hb either oral or parenteral virus Inoculation. The virus may be released from liver into blood and intestine vial the bile or just from the gut wall into feces, and hence it may be transmitted by both routes. PMID- 11498688 TI - [Molecular cloning and sequencing of 5' non-coding region of TTV-like mini virus]. AB - OBJECTIVE: To investigate the TTV- like mini virus (TLMV) infection in Chinese patients with non A-G hepatitis and to analyse sequence of 5' non-coding region (5'NCR)of TLMV. METHODS: TLMV DNA was detected by nested - PCR from 53 serum samples of non A-G hepatitis patients without TTV infection. PCR products were cloned and sequenced. RESULTS: TLMV DNA were positive in 37 of 53 patients (69.8%). The sequence of 5' NCR of 8 TLMV isolates was compared with that reported by Takahashi (GenBank Accession No. ab 026930-026931) and showed nucleotide identity between 64% - 83%. CONCLUSIONS: The results of this study indicated that TLMV infection is common in TTV negative patients with non A-G hepatitis. The 5' NCR sequence of TLMV showed a great degree of diversity. The pathogenicity of TLMV and its relationship to non A-G hepatitis need to be confirmed and further study is required. PMID- 11498689 TI - [Selective inhibition of HSV-1 DNA and protein synthesis by antiviral antibiotic 17997]. AB - OBJECTIVE: To study the effects of 17997 on the HSV-1 DNA and protein synthesis. METHODS: DNA synthesis was determined by isopycnic separation of cellular and viral DNA in CsCl gradients centrifugation. Protein synthesis was determined by SDS-PAGE and Western blot. RESULTS: The inhibition rates of 0.5 micromol/L, 1.0 micromol/L, 2.0 micromol/L and 4.0 micromol/L of 17997 on HSV-1 DNA synthesis were 63.1%,74. 1%, 93.9% and 100%, respectively. 4.0 micromol/L of 17997 did not inhibit VERO cell DNA synthesis. The above concentrations of 17997 did not show any inhibitory effects on cellular protein synthesis, but selectively inhibited on viral protein synthesis. CONCLUSIONS: 17997 selectively displayed inhibitive effects on HSV-1 DNA and protein synthesis. PMID- 11498691 TI - [Study on the inhibitory effect of antisense phosphorothioate oligodeoxynucleotide on coxsackie virus B replication in vitro]. AB - OBJECTIVE: To investigate the antiviral effect of antisense phosphorothioate oligodeoxynucleotide (AODN) on CVB3 replication in vitro. METHODS: In this study, 21 mer antisense phosphorothioate oligodeoxynucleotide which complemented to nt 581-601 in 5' NCR of Coxsackie Virus B3 (CVB3) RNA was used to investigate antiviral activity in Vero cells as a specific inhibitor of CVB3 replication. Specific AODN of CVB3 RNA, sense oligodeoxynucleotide (SODN) and randomized nonsense-sequence RODN were synthesized for effect comparison among them. Vero cells infected by CVB3 were transfected with different concentrations of AODN mediated by lipofectamine reagent, so that the cells could take in more AODN. Cell control and viral control were set up. The inhibitory effect of AODN on CVB3 replication was evaluated with a number of variables, including inhibitory rates of cytopathic effect (CPE), cell survival rates by MTT assay, inhibitory rates of CVB3 antigen by ELISA, inhibitory rates of RNA by dot blotting and 50% tissue culture infective dose (TCID50). RESULTS: The specific AODN could significantly inhibit CPE of CVB3-infected Vero cells, decrease the production of antigen and RNA of CVB3 and viral titers, and increase cell survival rates, in a dose dependent manner. The strongest inhibitory effect appeared at 48 hours after transfection, the most effective concentration of AODN was 10 micromol/L. On the other hand, 10 micromol/L SODN also showed weaker inhibitory effect on CPE of CVB3-infected cells, but no antiviral effect of 10 micromol/L RODN was shown. AODN showed no-inhibition on HSV-1 replication, but inhibited some enteroviruses such as CVB3, Polio-1 and Echo-6 to some extent. CONCLUSIONS: These results indicate that nt 581-601 in 5' NCR of CVB3 RNA may play an important role in regulating CVB3 replication and AODN may have inhibitory effect on CVB3 replication. PMID- 11498690 TI - [The effects of hepatitis G virus infection on clinical features and liver pathologic lesions of chronic hepatitis C]. AB - OBJECTIVE: To explore the clinical and pathological effect of hepatitis G virus infection on chronic hepatitis C. METHODS: Detecting HGV-RNA by reverse transcription-polymerase chain reaction from serum samples of 53 chronic hepatitis C through patients as diagnosed by liver biopsy. The clinical and pathologic features of the patients with positive HGV-RNA were compared with those of patients with negative HGV-RNA. RESULTS: The results showed that 15 patients (28.3%) were positive for serum HGV-RNA and there were no significant differences in clinical manifestations, biochemical indexes, HCV-RNA positive rates and the liver pathologic lesions between these HGY RNA positive and negative groups (P > 0.05). CONCLUSIONS: These data indicate that HGV coinfection does not affect the liver lesions and HCV replication of chronic hepatitis C. PMID- 11498692 TI - [Cloning and expression of TTV gene fragment in E. coil and identification of the expressed protein]. AB - OBJECTIVE: To construct a recombinant expression vector containing TTV gene fragment and identify the expressed protein. METHODS: Based on computer-aided studies, TTV ORF12 gene fragment coding protein of antigenicity was amplified by PCR from the patient serum with non-A-G hepatitis. Then the gene fragment was integrated into the multiple cloning site of pQE-30 and expressed in E coil. The expressed protein was identified by ELISA. RESULTS: The relative molecular weight of the expressed protein matches the predication and its yield was up to 22.33% of total bacterial proteins of E. coli The expressed protein was detected by ELISA. CONCLUSIONS: The recombinant vector has been constructed successfully and the expressed protein possesses antigenicity. PMID- 11498693 TI - [Establishment of a Hantaan virus infected mouse neuron cell model]. AB - OBJECTIVE: To establish a neuron cells model for Hantaan virus (HTNV) infection by using the cultures of cortical neurons of mouse embryo. Changes and mechanism of the damage in neurons after HTNV infection were studied. METHODS: The cortical neurons of 16-19 day old mouse embryo were cultured and infected with HTNV (A9 strain). Indirect immunofluorescence test (IFT), MTT colorimetric analysis and immunohistochemistry assay (IHCA) were used to detect immunofluorescent antibody reaction of neurons, growth and development of the neurons. RESULTS: In neurons of HTNV infected mouse embryo, continually cultured for one week, lFT antigen was detected positive, the activity of the infected neurons was reduced obviously and the growth of neurons was obstructed. CONCLUSIONS: Direct infection of HTNV on the cultured neurons was identified, indicating that mouse embryonic critical neurons is one of the sensitive target cells for HTNV infection. PMID- 11498694 TI - [Detection for anti-Hantavirus IgM in patient serum with silver enhanced dot immunogold filtration assay]. AB - OBJECTIVE: To establish a more rapid and simple method for detecting anti Hantavirus IgM in patient' s serum. METHODS: A silver enhanced dot immunogold filtration assay (SEDIGFA) was developed by coating the nucleocapsid protein of Hantavirus on nitrocellulose (NC) membrane, adding colloidal gold labeled sheep anti-human IgM antibody and the silver enhancer. The whole procedure can be finished within 10 minutes. RESULTS: The coincidence rate of the results of serum samples simultaneously operated with ELISA and SEDEIGFA was 98% for anti Hantavirus IgM. CONCLUSIONS: SEDIGFA is more simple and rapid for application to small medical laboratories. It can be applied for early diagnosis of hemorrhagic fever with renal syndrome (HFRS). PMID- 11498695 TI - [Establishment of in situ hybridization technique for detection of parvovirus B19 and its clinical application]. AB - OBJECTIVE: To establish in situ hybridization (ISH) technique to detect parvovirus B19 and to know the viral location in cardiac tissue of congenital heart diseases (CHD). METHODS: ISH method was established with randomly primed, degoxigenin labeled B19 DNA probe of 1112 bp long corresponding to parvovirus B19 capsid protein VP1 genome. RESULTS: Dot blot hybridization showed that B19 DNA was positive with the sensitivity of 0.1 pg/microl, while CMV, HSV, TOX DNA were all negative. 7 out of 66 CHD cardiac tissues were B19 DNA positive by using ISH, while in 38 cases of control group were all negative. B19 DNA positive signal was found in the nucleus of cardiac cell. CONCLUSIONS: We have established ISH to detect B19 DNA that has high sensitivity (0. 1 pg/microl) and specificity and have found that B19 DNA is located the nucleus of cardiac cell. PMID- 11498696 TI - [An experimental study of rat infected with human cytomegalovirus AD169 strain]. AB - OBJECTIVE: To study the sensitivity of rat to human cytomegalovirus (HCMV) infection and to establish a stable animal model. METHODS: Two kinds of rat experiments were carried out: 1) 30 SD rats were divided into three groups randomly, they were virus inoculated group, inactivated virus inoculated group and the normal rat group respectively; 2) 40 Wistar rats were randomly divided into two groups, they were the virus inoculated group and the inactivated virus inoculated group. All the rats were inoculated through tail vein. 90 d later, the rat's tissue lesions were observed by pathological techniques, HCMV antigen was detected by immunohistochemical method and the HCMV DNA was analyzed by in situ hybridization. RESULTS: The extensive pathological damages in tissues of HCMV infected rats were observed. Meanwhile, the viral antigen and viral DNA were also demonstrated in many tissues of HCMV infected rats. CONCLUSIONS: Rat is sensitive to infection of HCMV strain, but, as an animal model study, we still have a lot of works to do. PMID- 11498697 TI - [Study on typing of respiratory syncytial viruses]. AB - OBJECTIVE: To determine the epidemiologic patterns of subgroups A and B and genotypes of respiratory syncytial viruses (RSV) during two uncontinuously epidemic years from 1990 to 1991 and 1997 to 1998 in Beijing. METHODS: Nasopharyngeal secretion (NPS) samples of RSV positive or BSV isolates tested by indirect immunofluorescent (IIF) assay were classified into subgroups A and B. Also, isolates of RSV were divided into at least six different lineages, designated NPl-6, by restriction mapping of the N gene. NP l, 3 and 6 were given by subgroup B isolates, while NP2, 4 and 5 were given by subgroup A isolates. Strains of subgroup A were further subdivided into six lineages SHL 1-6 on the basis of the SH gene sequence. SH lineages were closely related to each other and to NP 1-6. Strains of SHL 1,3, and 4 were closely related and belonged to NP2, SHL 2 and 6 to NP4 and SHL5 to NP5. RESULTS: Of 145 RSV NPS samples from the winter 1997 to the spring 1998, 83(57.2%) were subgroup B strains, 62(42.8%) were sub-group A strains, the rate of occurrence of subgroup A to B strains was about 1:1.3. Two of 10 isolates of 1997-1998 offered by viral department of research institute of Pediatrics were subgroup A strains, whereas 8 were subgroup B strains, the rate of occurrence of subgroup A to B strains was 1:4. Eight subgroup A strains of 10 isolates from the 1990-1991 epidemic were dominant, the proportion of subgroup A to B strains was 4:1. of 10 BSV isolates in 1997-1998, all 2 subgroups A strains gave N gene fragment restriction pattern NP4, and fell into SH lineage SHL2, whereas 8 subgroup B strains all belonged to NP3. All 8 subgroup A isolates from the 1990-1991 epidemic gave pattern NP4, and fell into SHL2, while 2 subgroups B strains all belonged to NP3. The classification of subgroups A and B deduced from NP patterns was corresponded to the definition of these subgroups by monoclonal antibodies. CONCLUSIONS: These observations confirmed that subgroups A and B or multiple lineages of BSV co-circulated in Beijing, but different genomic types predominated each year. Moreover, very similar viruses were isolated up to more than 5 years ago, indicating that despite apparent diversity of the subgroup A strains, the separate lineages might be relatively stable. PMID- 11498698 TI - [Typing of enteric adenoviruses in feces of infants with diarrhea]. AB - OBJECTIVE: To study the infection condition of adenovirus types in infants with diarrhea in Shenzhen. METHODS: 114 fecal samples from diarrhea infants were tested by PCR. The amplified products were analyzed by Taq I and Rsa I endonucleases. RESULTS: The positive rate of adenovirus DNA in fecal samples was 14% (16/114). The amplified products typing procedure started with Taq I endonuclease to identify the type of adenovirus, 11 cleaved by Taq I yielded two fragments of 191 bp and 110 bp, the positive rate of enteric adenovirus was 9.65% (11/114). Followed by Rsa I cleavage, two amplified products yielded 256 bp and 45 bp fragments (Ad40), the positive rate of Ad40 was 1. 75% (2/114), nine amplified products yielded 211 bp and 90 bp fragments (Ad4l), the positive rate of Ad41 was 7.89%(9/114). CONCLUSIONS: The positive rates of the adenovirus types 40 and 41 were 9.65% in feces of infants with diarrhea, and that of the other types of adenovirus was 4.35%. PMID- 11498699 TI - [Expression of glycoprotein gene of the rabies virus 3aG strain by E-3 deleted adenovirus recombinant]. AB - OBJECTIVE: To analyze Chinese rabies virus vaccine strain 3aG glycoprotein (GP) gene and further produce GP by E3-deleted human adenovirus recombinant. METHODS: Chinese rabies virus vaccine strain 3aG glycoprotein gene was cloned by RT-PCR and its sequence was determined by DNA sequencing. Cotransfection was performed to obtain adenovirus recombinant. The expressed glycoprotein was examined by ELISA and its immunogenicity was evaluated by testing neutralizing antibody level of mice inoculated with the recombinant virus. RESULTS: DNA sequencing showed that the open reading frame of GP gene contains 1,575 nucleotides and five of the deduced amino acids are different from the previous report. The recombinant adenovirus containing GP gene in E3 region was obtained by cotransfecting 293 cells and rounds of plaque purification. ELISA assay demonstrated that the GP gene can be efficiently expressed and rapid fluorescent focus inhibition test (RFFTT) showed it can also elicit GP specific neutralizing antibody. CONCLUSIONS: Chinese rabies virus vaccine strain 3aG glycoprotein was successfully expressed by E3-deleted human adenovirus recombinant and specific neutralizing antibody can be elicited in mice after immunization by the recombinant. PMID- 11498700 TI - [Novel antibiotics active against Gram-positive bacteria]. PMID- 11498701 TI - [The new decade of aminoglycosides: Their use in clinical practice]. PMID- 11498702 TI - [Cefepime in the treatment of osteomyelitis caused by Gram negative bacilli]. AB - We conducted a prospective, randomized, open-label trial to evaluate the efficacy and tolerability of cefepime in the treatment of osteomyelitis caused by Gram negative bacilli. Hospitalized patients with diagnosis of osteomyelitis due to Gram-negative bacilli susceptible to cefepime were elegible for enrollment. Cefepime was administered intravenously or intramuscularly (2 g every 8 or 12 hours). Microorganisms were considered susceptible to cefepime when the MIC was <8 mg/l. Forty-five patients with bone infections were enrolled, forty-three with osteomyelitis (22 chronic and 21 acute) and two with arthritis. In the per protocol analysis 42 patients were evaluated: 30 (71.4%) were cured. In the intent to treat analysis 45 patients were evaluated: 33 (73.3%) were cured. Our trial suggests that cefepime is as effective as other modern parenteral beta lactam antibiotics in the treatment of osteomyelitis due to Gram-negative bacilli. PMID- 11498703 TI - [Influence of serious infections due to Gram-negative bacteria on the hospital economy]. AB - Nosocomial infections due to Gram-negative bacteria are very important since they are associated with high morbidity and high hospital costs. A prospective study of 250 inpatients was carried out, 200 of whom had Gram-negative bacterial infections. Patients were divided into groups of 50 according to the localization of the infection (urinary, surgical wound, respiratory tract and bacteremia), with a control group of 50 patients with similar characteristics but no infection. We calculated the cost for the different groups by multiplying the average length of hospital stay in days by the daily cost of the stay. Significant differences were observed in the average length of stay per patient according to the type of infection and how it was acquired. In terms of cost, nosocomial infection due to Gram-negative bacteria was 1,049,139 pesetas more expensive than community-acquired infection. The cost of the stay for patients with postsurgical infection due to Gram-negative bacteria was 1,108, 252 pesetas more expensive than for the group of control patients. Nosocomial infection due to Gram-negative bacteria is associated with a prolongation in hospital stay of 9 to 28 days, which is the factor that most reflects the cost that can be attributed to nosocomial infection. Consensual and protocolized measures which allow for better clinical management need to be developed. PMID- 11498704 TI - [Prospective, comparative study (1994-1998) of the influence of short-term prophylactic treatment with azithromycin on patients with advanced COPD]. AB - Despite the advances in therapy, chronic obstructive pulmonary disease (COPD) requires frequent hospital admissions due to acute exacerbations. We carried out a prospective randomized study of two groups of patients with COPD, one (n = 54) treated with azithromycin (500 mg/day) for three days every 21 days during the winter months, and a control group (n = 40) without treatment. A statistically significant reduction in the number of acute infectious episodes (187) and hospital admissions (22) was observed in the treated group versus the control group (249 and 45, respectively). A short prophylactic treatment course with azithromycin is a good alternative in the management of patients with severe, advanced COPD, and could lead to an improvement in social and healthcare costs PMID- 11498705 TI - [Profile of bacterial isolates and antimicrobial susceptibility: Multicenter study using a one-day cut-off]. AB - The frequency and antimicrobial susceptibility of bacterial isolates from the largest clinical samples collected in 19 Spanish hospitals were studied. A total of 523 strains were identified and grouped by sample. Blood stream: Staphylococcus coagulase-negative (41%) and Escherichia coli (19.7%); oxacillin resistance occurred in 44% of coagulase-negative strains, strains which were also resistant to nonbetalactam agents. All antimicrobial agents tested had good activity against E. coli, with the exception of penicillins (25 to 33% susceptible). Urine: E. coli (59.1%) and Enterococcus faecalis (15%); aminoglycosides and third generation cephalosporins were the most active compounds against E. coli, whereas penicillins and cotrimoxazole were the least active. E. faecalis isolates showed low rates of resistance to the antibiotics tested and no glycopeptide-resistant strains were detected. Skin and soft tissues: Staphylococcus aureus (24.1%), Pseudomonas aeruginosa (17.7.%); oxacillin resistance occurred in 15.8% of S. aureus strains and co-resistance to nonbetalactam agents was frequently observed among these strains. Ceftazidime susceptibility was elevated among P. aeruginosa (76.9%) and the most active agents were aminoglycosides (100% susceptibility). Lower respiratory tract: P. aeruginosa (21.4%) and Haemophilus influenzae (15.5%). Aminoglycosides (88.8 to 94.4%) and ceftazidime (72%) presented the highest susceptibility rates in P. aeruginosa. All the agents tested were very active against H. influenzae (89% susceptibility). Among Gram-positive cocci, no vancomycin and/or teicoplanin resistant strains were detected. PMID- 11498706 TI - [Evolution of antimicrobial susceptibility of Acinetobacter baumannii clinical isolates]. AB - Acinetobacter baumannii is a microorganism frequently implicated in colonization and infection in hospitalized patients. An increase of resistance has been observed in recent years making these infections difficult to treat. The in vitro activity of 24 antibiotics, 15 betalactam agents and nine nonbetalactams, was studied in 156 A. baumannii clinical isolates. The strains were collected from different clinical samples obtained from inpatients (92%) and 8% were from outpatients. Evolution of susceptibility from January 1995 to December 1997 was studied. MIC of the following antibiotics was determined by the agar dilution method: ampicillin, ticarcillin, piperacillin, ampicillin-sulbactam, amoxicillin- clavulanic acid, ticarcillin-clavulanic acid, piperacillin-tazobactam, cefotaxime, ceftazidime, cefepime, imipenem, meropenem, clavulanic acid, sulbactam, tazobactam, amikacin, gentamicin, tobramycin, ofloxacin, doxycycline, fosfomycin, rifampin, azithromycin and colistin. Low antimicrobial susceptibility was observed in most A. baumannii strains. Colistin, imipenem, meropenem and ampicillin-sulbactam showed the greatest susceptibility (100, 88.4, 88.4 and 84.6%, respectively). A. baumannii strains from inpatients showed a lower antimicrobial susceptibility than strains from outpatients, who showed a high percentage of susceptibility to most antibiotics. Rifampin and azithromycin showed certain in vitro activity against the most susceptible A. baumannii strains. A progressive decrease in susceptibility to most antibiotics was observed during the period studied. Carbapenem-resistant A. baumannii emerged in 1996 and increased in 1997. PMID- 11498707 TI - [Antibacterial activity of aminoglycosides against nosocomial strains of Acinetobacter baumannii isolated between 1990 and 1999]. AB - The antibacterial activity of selected aminoglycosides against 487 nosocomial strains of Acinetobacter baumannii isolated from Chilean hospitals between 1990 and 1998 was investigated. Plasmid profiles were determined for some resistant isolates. A high frequency of resistant strains was found for all aminoglycosides assayed, with values ranging from 47% for amikacin (1990-1992) to 88% for gentamicin (1993-1994). The cure of a plasmid greater than 30 kb was associated with the loss of amikacin, kanamycin and neomycin resistance in some isolates. This result suggests that plasmid may harbor the genes encoding for the combination of aminoglycoside-modifying enzymes AAC(6')I+APH(3'). Gentamicin resistance was not lost among cured strains, suggesting it may be encoded by genes located on the bacterial chromosome or transposons inserted on the chromosome. PMID- 11498708 TI - [Activity of imipenem and meropenem against strains of Acinetobacter baumannii isolated from various Chilean hospitals]. AB - The activity of two carbapenem compounds, imipenem and meropenem, against 447 strains of Acinetobacter baumannii isolated between 1990 and 1998 in different Chilean hospitals was determined. MIC ranges, MIC(50) and MIC(90), were determined by an agar dilution method. Similar antibacterial activities were observed for both antibacterials; however, a slight increase in the MIC(50) of imipenem and meropenem, and in the MIC(90) of meropenem was found among strains isolated from 1997-1998. Although A. baumannii remains susceptible to these antibiotics, the MIC(50) and MIC(90) have increased in recent years. PMID- 11498709 TI - [In vitro activity of ascorbic acid]. AB - The aim of this study was to determine the in vitro activity of ascorbic acid (vitamin C) against 43 Helicobacter pylori clinical isolates obtained from gastric biopsy taken by endoscopy. The MIC was determined by an agar dilution method. The ascorbic acid was included as two-fold dilutions in the agar to obtain a final concentration from 4000 mg/l to 62 mg/l. Brain-heart infusion agar supplemented with yeast extract and 10% fetal calf serum, 10(6) CFU/spot inoculum and microaerobic incubation for 2 to 5 days were used. MIC(50) 500 mg/l, MIC(90) 1000 mg/l and the range from 62 to 2000 mg/l. No differences in susceptible or resistant isolates were observed. Ascorbic acid showed good in vitro activity against the H. pylori clinical isolates tested in this study, which could explain some of the beneficial properties of vitamin C. PMID- 11498710 TI - [Resistance to antibiotics in clinical isolates of the genus Enterococcus]. AB - The purpose of our study was to determine the prevalence of resistance in 114 clinical strains of enterococci. Identification was made using PASCO dehydrated panels and by the conventional method of Facklam and found the following: 92.1% E. faecalis, 7% E. faecium and 0.9% E. avium. The antibiotic susceptibility was determined by the PASCO system, and the minimum inhibitory concentrations of vancomycin, teicoplanin, gentamicin and streptomycin were determined by the agar dilution method. All the strains were susceptible to vancomycin and teicoplanin. We detected resistance to ampicillin in 4.4% of isolates, to penicillin in 6%, to tetracycline in 41.2%, to rifampicin in 12.3%, to chloramphenicol in 23.7%, to erythromycin in 66.7%, to ciprofloxacin in 53.5%, to gentamicin to a high level in 43.9%, and to streptomycin to a high level in 57%. E. faecium was associated with high-level resistance to gentamicin. E. faecium was more resistant than E. faecalis to quinolones, rifampicin, penicillin and ampicillin, and showed similar susceptibility to tetracycline and chloramphenicol. However, E. faecalis was more resistant than E. faecium to aminoglycosides, mainly to gentamicin. PMID- 11498711 TI - [Five-year analysis of extrahospital consumption of betalactams in Spain]. AB - We carried out a qualitative and quantitative study to determine extrahospital consumption of both broad-spectrum penicillins and cephalosporins in Spain in the period 1993-1997. Penicillins were the most consumed group, followed by macrolides and cephalosporins. Units and value (ptas.) of penicillins and cephalosporins during these five years show interannual variations. Nevertheless, monthly analysis of consumption for both groups showed a stable tendency, with peaks in winter months and drops in the summer. Both penicillins (54.38%) and cephalosporins (27.71%) were prescribed mainly for upper respiratory tract infections. Men received more penicillins and cephalosporins than women (51.02% and 55.09%). Children under 11 years were the main group for consumption of both types of antibiotic, while patients aged 55-64 years were the group with the least consumption. PMID- 11498712 TI - [Stevens-Jhonson syndrome associated with vancomcyn]. PMID- 11498713 TI - (R)-methanandamide and Delta 9-THC as discriminative stimuli in rats: tests with the cannabinoid antagonist SR-141716 and the endogenous ligand anandamide. AB - RATIONALE AND OBJECTIVES: (R)-methanandamide (AM-356), a metabolically more stable chiral analog of the endocannabinoid ligand anandamide, was used as a representative of fatty acid ethanolamide CB1 receptor ligands to characterize the discriminative stimulus functions of anandamides. METHODS: Rats discriminated between 10 mg/kg (R)-methanandamide and vehicle administered IP 15 min prior to session onset. Another group of rats was initially trained to discriminate between 3 mg/kg Delta9-THC and vehicle given IP 30 min prior to session onset; for anandamide testing, the animals were retrained with 1.8 and 5.6 mg/kg Delta9 THC. A two lever operant methodology (FR10) was used. RESULTS: Delta9-THC was more potent than (R)-methanandamide at both 15 and 30 min post-injection, irrespective of the training drug used. Additional tests with 10 and 18 mg/kg (R) methanandamide suggested that the effects were declining by 1 h. The cannabinoid antagonist SR 141716 (0.3 and 1 mg/kg) produced rightward shifts in the Delta9 THC dose-response curve for Delta9-THC-appropriate responding and for (R) methanandamide-appropriate responding (surmountable antagonism). SR-141716 (0.3 and 1 mg/kg) antagonized the ability of (R)-methanandamide to occasion either Delta9-THC-appropriate responding or (R)-methanandamide-appropriate responding. This antagonism was surmountable only at a dose of 0.3 mg/kg SR-1421716 in the (R)-methanandamide-trained rats. SR-141716 did not antagonize the rate-decreasing effects of (R)-methanandamide in either the Delta9-THC or the (R)-methanandamide trained rats. Response suppression precluded testing doses higher than 30 mg/kg (R)-methanandamide. Tests with SR-141716 (1 and 10 mg/kg) alone resulted in <3% Delta9-THC-appropriate responding. With 10 mg/kg SR-141716, response rate was significantly lower as compared to the rate observed during a vehicle test. Tests with anandamide (10 and 18 mg/kg) resulted in 41% and 85% (R)-methanandamide appropriate responding at a 3-min pre-treatment time, but in a maximum of 15% (R) methanandamide-appropriate responding at a longer (15 min) pre-treatment time. In the Delta9-THC (1.8 and 5.6 mg/kg) trained rats, anandamide never produced more than about 20% Delta9-THC-appropriate responding. CONCLUSION: The results add to a growing body of evidence indicating that there are both similarities and dissimilarities between classical cannabinoids such as THC and endogenous fatty acid ethanolamides. PMID- 11498714 TI - Effects of contextual or olfactory cues previously paired with morphine withdrawal on behavior and pain sensitivity in the rat. AB - RATIONALE: Pavlovian conditioning processes have been accorded an important role in maintaining persistent opiate administration. At least one locus for this contribution is during opiate withdrawal. These experiments studied the contribution of Pavlovian conditioning processes to morphine withdrawal. OBJECTIVES: To determine whether exposure to a distinctive context or odor paired with morphine withdrawal would provoke a withdrawal syndrome, defensive behaviors (e.g., freezing) and pain modulatory (e.g., hypoalgesia) responses similar to those produced by exposure to stimuli signaling other sources of aversive stimulation (e.g., footshock), or whether both withdrawal and fear-like responses would be provoked. METHODS: Rats were used in four experiments to study the effects on defensive behavior and pain sensitivity of naloxone-precipitated morphine withdrawal or exposure to a distinctive context or odor previously paired with such withdrawal. RESULTS: Injection of 2.5 mg/kg naloxone in morphine dependent rats precipitated a withdrawal syndrome characterized by whole body shaking, diarrhea, ptosis, and postural abnormalities (experiment 1). Exposure to either a distinctive context (experiment 2) or odor (experiments 3) previously paired with morphine withdrawal provoked the species-typical defense response of freezing but not signs of withdrawal. Exposure to an odor previously paired with morphine withdrawal also provoked hypoalgesia in the formalin test, which was mediated by activity at opioid receptors (experiment 4). CONCLUSIONS: These results show that opiate withdrawal supports the conditioning of defensive and hypoalgesic responses consistent with the arousal of a fear motivational system. The emergence of fear in these experiments, and the relationship between the freezing observed here and the learned avoidance and suppression observed in other withdrawal conditioning preparations, is discussed with reference to dual representation accounts of Pavlovian conditioning. PMID- 11498715 TI - Effects of smoking/nicotine on performance and event-related potentials during a short-term memory scanning task. AB - RATIONALE: Nicotine absorbed from cigarette smoke shortens reaction time (RT) in a wide variety of cognitive tasks. However, relatively few studies have tried to isolate the specific stage(s) of information processing affected by smoking/nicotine. OBJECTIVE: The present study was designed to investigate the effect of smoking/nicotine on the short-term memory (STM) scanning stage of information processing in minimally abstaining smokers. Both RT and event-related potentials (ERPs) were measured. METHODS: A Sternberg-type STM-scanning task was performed before and after smoking each of two cigarettes. One cigarette had a 0.05-mg nicotine yield ("denicotinized") and the other had a 1.1-mg yield ("nicotine-yielding"). On each trial, either 2, 3, or 4 consonants were displayed as a memory set. After a brief interval, a single probe consonant was displayed. If the probe was in the memory set (positive probe) a right button press was required, and if the probe was not in the memory set (negative probe) the left button was pressed. RESULTS: Smoking the nicotine-yielding cigarette but not the denicotinized cigarette shortened RT. However, memory-scanning speed, as estimated from the increase in RT as a function of increasing set size, was not differentially affected by the two types of cigarettes. For the ERPs, smoking the nicotine-yielding but not the denicotinized cigarette (a) reduced N200 latency to both the memory-set stimuli and negative probes, (b) increased N200 amplitude to negative probes and P300 amplitude to both types of probes, and (c) produced a sustained negative shift in memory-set ERP amplitude beginning around 600 ms post stimulus. CONCLUSION: While smoking/nicotine shortened probe RT, it did not affect the speed of STM scanning. Moreover, the ERP-latency effects obtained for the probes were small relative to the effects of smoking/nicotine on RT, suggesting that smoking/nicotine shortens RT primarily by affecting response related processes. PMID- 11498716 TI - Facilitation and inhibition of retrieval in two aversive tasks in rats by intrahippocampal infusion of agonists of specific glutamate metabotropic receptor subtypes. AB - RATIONALE: The generic antagonist of glutamate metabotropic receptors (mGlus), MCPG, blocks retrieval of inhibitory avoidance when infused into the CA1 area of rat hippocampus. It was considered important to study the effect of agonists of different types of mGlus on retrieval both of this task and of a related one, contextual fear. OBJECTIVES: To measure the effect of three mGlu agonists (3HPG, which is selective to mGlu1; LCCG, which binds to mGlu2 and mGlu3; and LAP-4, which binds to mGlu4 and mGlu6), infused bilaterally into CA1, on the retrieval of one-trial inhibitory avoidance and contextual fear in rats. METHODS: Rats bilaterally implanted with cannulae in the CA1 region of the dorsal hippocampus were trained in one-trial step-down inhibitory avoidance or in a contextual fear task and tested for retention 24 h later. The drugs 3HPG, LCCG and LAP-4 were infused into CA1 at different concentration levels 10 min before retention testing. In addition, we studied the effect of these drugs on locomotor and exploratory activity measured in an open field, and on pro- and anti-conflict behaviour in an elevated plus-maze. RESULTS: 3HPG hindered, and LCCG and LAP-4 enhanced, retrieval of the two tasks. In all cases the effects were dose dependent. The drugs had no effects on open field or plus maze behaviour. CONCLUSIONS: Retrieval of one-trial inhibitory avoidance and of contextual fear is regulated by mGlus in the CA1area of the rat hippocampus. The results suggest that mGlu2s, mGlu3s, mGlu4s and mGlu6s are necessary for retrieval and that mGlu1s play an inhibitory role. The effects are not explainable by nonspecific influences on locomotor or exploratory activity or anxiety levels. PMID- 11498717 TI - L-dopa induces dyskinesia in normal monkeys: behavioural and pharmacokinetic observations. AB - RATIONALE: L-Dopa induces dyskinesias during the treatment of Parkinson's disease and also in primates with nigrostriatal lesions produced by MPTP, but it is claimed that L-dopa does not provoke dyskinesia in humans or monkeys with an intact or mildly damaged nigrostriatal system. OBJECTIVES: This study assessed the behavioural and pharmacokinetic effects of chronic oral administration of L dopa plus carbidopa alone, or with co-administration of the peripheral COMT inhibitor entacapone, to normal macaque monkeys. Repeated high dose L-dopa administration was shown to induce marked dyskinesias in monkeys with an intact nigrostriatal system, and the threshold for dyskinesia expression was increased by peripheral catechol-O-methyltransferase inhibition with entacapone. METHODS: Six groups of normal macaque monkeys (n=8 per group; Macaca fascicularis) were treated with L-dopa (20, 40 or 80 mg/kg) plus carbidopa (5, 10 or 20 mg/kg) with or without the catechol-O-methyltransferase inhibitor entacapone (20, 40 or 80 mg/kg), or with entacapone alone (80 mg/kg), by oral administration once daily for 13 weeks. RESULTS: Eleven of 16 animals receiving high dose L-dopa (80 mg/kg plus carbidopa 20 mg/kg PO with or without entacapone 80 mg/kg PO for 13 weeks) gradually developed reproducible and idiosyncratic combinations of chorea, athetosis and dystonia maximal at 60-100 min after L-dopa administration, which progressively intensified over the course of the study. The dyskinesias observed were similar in type and distribution to L-dopa-induced dyskinesia observed in patients with Parkinson's disease and in MPTP-treated primates. The occurrence of dyskinesia correlated with plasma concentrations of L-dopa, with animals displaying the most severe dyskinesias having significantly higher plasma concentrations of L-dopa one hour after dosing than animals with mild or moderate dyskinesia or no dyskinesia. Co-administration of entacapone with L-dopa plus carbidopa significantly lowered peak plasma concentrations of L-dopa and this was reflected by a decrease in the severity of dyskinesias, with only one animal receiving entacapone and high dose L-dopa plus carbidopa showing severe dyskinesia, while four receiving high dose L-dopa plus carbidopa alone did so. CONCLUSIONS: These results show for the first time that chronic oral L-dopa administration can provoke dyskinesias in primates independently of nigrostriatal damage, and that this effect is dose related. PMID- 11498718 TI - Intravenous self-administration of the cannabinoid CB1 receptor agonist WIN 55,212-2 in rats. AB - RATIONALE: Delta9-tetrahydrocannabinol (Delta9-THC), the main psychoactive ingredient of marijuana, as well as synthetic cannabinoid (CB1) receptor agonists, has led to negative or equivocal results when tested with the intravenous self-administration procedure, the best validated behavioural model for evaluating abuse liability of drugs in experimental animals. We recently reported, however, that the synthetic CB1 receptor agonist WIN 55,212-2 is intravenously self-administered by drug-naive mice and that its self administration is blocked by the cannabinoid CB1 receptor antagonist SR 141716A. OBJECTIVE: To assess a reliable model of cannabinoid intravenous self administration in rats. Long Evans male rats were allowed the opportunity to self administer WIN 55,212-2 at doses ranging from 6.25 to 50 microg/kg per injection, under a fixed-ratio 1 (FR1) schedule of reinforcement and nose-pokes as the operant responses. The effect of either a change in the unit drug dose available or a pretreatment with the specific CB1 receptor antagonist SR 141716A were then investigated (maintenance phase). Finally, the extinction of the self administration behaviour was evaluated. RESULTS: Response rate depended on the drug dose available, with maximum rates occurring at 12.5 microg/kg per injection. Response rate increased following pretreatment with the specific CB1 receptor antagonist, SR 141716A. Moreover, operant behaviour rapidly extinguished following both the substitution of saline or vehicle for cannabinoid and the disconnection of the drug delivery pumps. CONCLUSION: Rats will intravenously self-administer the synthetic CB1 receptor agonist WIN 55,212-2 under specific experimental conditions, thus allowing further investigation of the neurobiological mechanisms underlying cannabinoid-taking behaviour. PMID- 11498719 TI - Comparative effects of melatonin, zolpidem and diazepam on sleep, body temperature, blood pressure and heart rate measured by radiotelemetry in Wistar rats. AB - RATIONALE: The role of melatonin (MLT) in mediating the sleep-wake cycle has been previously suspected of indicating that this substance could be a candidate for a new generation of hypnotics. OBJECTIVES: We investigated whether MLT acted as a sleep promoter or a modulator of sleep temporal timing related to cardiovascular and body temperature (Tb) adaptations to sleep induction. The pharmacological effects of MLT on sleep were compared with zolpidem (ZP) and diazepam (DZ). METHODS: The radiotelemetry system was used to record the electrocorticogram [slow wave sleep (SWS), paradoxical sleep (PS)], Tb, blood pressure and heart rate in six Wistar rats. DZ (3 mg/kg and 6 mg/kg), ZP (1, 3, 5 and 10 mg/kg) and MLT (2.5 and 5 mg/kg) were delivered intraperitoneally during light (L) and dark (D) periods. RESULTS: MLT increased the number of sleep cycles (L: 30%, D: 110%) and total duration (P<0.05) of PS (L: 70%, D: 150%). In return, ZP (10 mg/kg) presented no effect during L but increased total (40%) and mean duration (37%) of SWS during the D period. DZ modified mean duration of SWS (L: -27%, D: +26%) and increased total duration of SWS (+47%). ZP and DZ induced a more pronounced decrease in Tb than MLT but only DZ induced tachycardia and hypertension. CONCLUSIONS: We showed that MLT could not promote sleep and its cardiovascular adaptations despite hypothermia, but modulated the period of ultradian sleep cycles. DZ and ZP promoted sleep and induced hypothermia during the D period. Only DZ disrupted sleep architecture and induced adverse effects on cardiovascular parameters. PMID- 11498720 TI - Temporal factors affecting cocaine-opioid interactions: a cocaine drug discrimination study in rats. AB - RATIONALE: Increasing concomitant abuse of cocaine and morphine-like opioids has prompted a number of studies aimed at understanding how these drugs interact. OBJECTIVE: The present study was designed to determine if variations in opioid pretreatment time would affect how mu opioid agonists interact with cocaine. METHODS: Rats were trained to discriminate 10 mg/kg cocaine from saline. One group of subjects (n=6) received morphine (5.6 mg/kg) 0.5 h, 1 h or 4 h prior to cumulative doses of cocaine (0.1-17.8 mg/kg). These pretreatment times were selected to overlap with states of acute opioid tolerance (approximately 1 h) or acute opioid dependence (approximately 4 h) as demonstrated by previous studies. A second group (n=6) was administered naloxone (0.3 mg/kg) 5 min prior to cumulative cocaine doses, with or without a 4-h morphine (5.6 mg/kg) or methadone (3.0 mg/kg) pretreatment. In a third procedure, the same subjects used in the second experiment were also tested for time-dependent changes in the analgesic effect of morphine using a hot-plate assay. RESULTS: Morphine pretreatment 1 h prior to assessment of the cocaine dose-response function significantly enhanced the discriminative stimulus effects of cocaine. However, neither 0.5-h or 4-h morphine pretreatment had any effect. In contrast, when naloxone was administered 4 h following either morphine or methadone and 5 min prior to assessment of the cocaine dose-response curve, the discriminative stimulus effects of cocaine were significantly attenuated. In assessing morphine-induced analgesia, paw-lick latency was significantly longer at 1 h and shorter at 4 h following morphine administration. CONCLUSIONS: The results illustrate the importance of temporal parameters for interactions between cocaine and mu opioid agonists. PMID- 11498721 TI - Physiological, subjective and reinforcing effects of oral and intravenous cocaine in humans. AB - RATIONALE: There is little comparative information on the qualitative similarity, relative potency and reinforcing effects of oral cocaine versus cocaine administered via other routes. METHODS: The present study used a within-subject, double-blind, double-dummy design to compare the physiological, subjective and reinforcing effects of placebo and oral (62.5, 125, 250 mg/70 kg) and intravenous (IV) (12.5, 25, 50 mg/70 kg) cocaine in volunteers with histories of cocaine abuse. RESULTS: Cocaine produced dose-dependent increases on heart rate and blood pressure, with effects lasting longer after oral than IV cocaine. Subjective ratings (e.g., "rush," "drug effect," "liking") were qualitatively similar and dose-dependently increased after oral and IV administration, and the duration of effects was similar under both routes. On a money versus drug choice measure of reinforcement, the monetary amounts at which participants chose drug over money increased as a function of cocaine dose under both routes of administration. At doses that produced comparable subjective, physiological, and reinforcing effects, oral cocaine was not identified as cocaine as frequently as IV cocaine. Across measures, the data suggested that IV cocaine was approximately 10 times more potent than oral cocaine. CONCLUSIONS: Overall, the results of this study support qualitatively similar effects of oral and IV cocaine and suggest that oral cocaine may be an effective tool for studying cocaine's effects in human laboratory studies. PMID- 11498722 TI - Age-related differences in sensitivity to the antinociceptive effects of opioids in male rats. Influence of nociceptive intensity and intrinsic efficacy at the mu receptor. AB - RATIONALE: Despite the widespread popularity of opioid analgesics, significant differences in the potency and effectiveness of these drugs are often observed across age groups. OBJECTIVES: The purpose of this investigation was to examine age-related differences in sensitivity to the antinociceptive effects of mu opioids and to identify the conditions under which these differences are most apparent. METHODS: In a warm-water tail-withdrawal procedure, young (3 months) and aged (24 months) male rats were habituated to restraint and the latencies to remove their tails from 50 degrees C (low nociceptive intensity) and 55 degrees C (high nociceptive intensity) water were measured. Opioids possessing a range of intrinsic efficacy at the mu receptor (morphine, levorphanol, buprenorphine, butorphanol, nalbuphine, nalorphine) were examined. RESULTS: Young and aged rats were equally sensitive to the antinociceptive effects of morphine, levorphanol, and buprenorphine when tested at the low nociceptive intensity. When these drugs were tested at the high nociceptive intensity, differences between the two age groups became apparent, such that aged rats were significantly more sensitive to the antinociceptive effects of these drugs than young rats. Differences between age groups were most apparent when butorphanol, nalbuphine, and nalorphine were tested, in that each of these drugs produced maximal levels of antinociception in aged rats under conditions in which they failed to produce antinociceptive activity in young rats. Under conditions in which lower efficacy opioids failed to produce antinociceptive activity in young rats, they antagonized the effects of morphine in drug combination tests. CONCLUSIONS: These data may be taken as evidence that aged male rats are more sensitive to the antinociceptive effects of mu opioids than young male rats, and that age-related differences in opioid sensitivity are most apparent when lower efficacy opioids and higher nociceptive intensities are employed during behavioral testing. PMID- 11498723 TI - Opposite effects of nefazodone in two human models of anxiety. AB - RATIONALE AND OBJECTIVES: To explore further the role of serotonin (5-HT) in anxiety, the effects of the 5-HT reuptake inhibitor and 5-HT2A receptor antagonist nefazodone (NF) were measured in two human models of anxiety. METHODS: Twenty-nine adult healthy volunteers of both sexes underwent conditioning of skin conductance responses (CSCR) to a tone associated to an aversive white noise. Another 34 subjects performed a simulated public speaking (SPS) task, seemingly related to unconditioned fear. In both tests, subjective states were evaluated through the visual analogue mood scale (VAMS) and a bodily symptoms scale (BSS). In each experiment, subjects were randomly divided into three groups, which received 100 mg NF, 200 mg NF or placebo under double-blind condition. RESULTS: In the CSCR test, NF decreased the number of spontaneous fluctuations of skin conductance (F=4.94; df=2,26; P=0.015). In addition, the increase in VAMS anxiety factor induced by the conditioning task was attenuated by NF (F=11.11; df=2,26; P<0.001). In contrast, the rise of VAMS anxiety induced by SPS was enhanced by NF (F=8.01; df=2,31; P=0.002). CONCLUSIONS: These results indicate that NF decreases conditioned anxiety, while enhancing unconditioned fear. Since the effects of NF may be due to impairment of 5-HT neurotransmission, consequent to overstimulation of autosomic 5-HT1A receptors and blockade of post-synaptic 5-HT2A receptors, the present results support the hypothesis that 5-HT facilitates conditioned anxiety, which may be related to generalised anxiety disorder, while inhibiting unconditioned fear, supposedly related to panic disorder. PMID- 11498725 TI - Once weekly administration of nicotine produces long-lasting locomotor sensitization in rats via a nicotinic receptor-mediated mechanism. AB - RATIONALE: Chronic nicotine administration results in dynamic changes in neuronal function, expressed as behavioral sensitization in animals and addiction in smokers. OBJECTIVES: The present study was undertaken to determine whether once weekly nicotine injection produces sensitization to the locomotor-activating properties of nicotine as a result of nicotinic receptor activation. METHODS: Once weekly for 6 weeks, rats were administered (s.c.) two saline injections or saline and nicotine (0.35 mg/kg), and locomotor activity was monitored. Rats remained in the home cage for 21 days, and subsequently were injected with the appropriate treatment to determine whether sensitization persisted. Rats were also injected with saline or mecamylamine (1.2 mg/kg) followed by saline or nicotine once weekly for 6 weeks to determine the effect of mecamylamine and whether it inhibited nicotine-induced hyperactivity. A separate group was injected with saline and nicotine once weekly for 4 weeks; on week 5, mecamylamine and nicotine were administered to determine whether mecamylamine inhibited the expression of sensitization. Separate groups were injected with mecamylamine and nicotine once weekly for 5 weeks or 6 weeks; on week 6 or week 9, respectively, saline and nicotine were injected to determine whether mecamylamine inhibited the initiation of sensitization. RESULTS: Sensitization to the locomotor-activating properties of nicotine developed following four nicotine injections across a 28-day period and persisted following 21 days of no drug treatment. Mecamylamine did not alter activity but attenuated both the initiation and expression of sensitization. CONCLUSIONS: Nicotinic receptor activation following once-weekly nicotine administration produces long-lasting behavioral sensitization, suggesting that even infrequent nicotine exposure initiates neuroadaptive processes associated with nicotine addiction. PMID- 11498724 TI - Different types of GABA(A) receptors may mediate the anticonflict and response rate-decreasing effects of zaleplon, zolpidem, and midazolam in squirrel monkeys. AB - RATIONALE: The role of different types of GABA(A) receptors in mediating anticonflict and response rate-decreasing effects of benzodiazepines in primate species is not known. OBJECTIVE: To examine the behavioral effects of the benzodiazepine-site, GABA(A) agonists zolpidem, zaleplon, and midazolam in the presence of two antagonists, flumazenil and beta-carboline-3-carboxylate-t-butyl ester (beta-CCt) in squirrel monkeys. METHODS: Two schedules of operant responding were used: (1) a multiple fixed-ratio (FR) schedule of food presentation involving punished and nonpunished behavior, and (2) an FR schedule of stimulus shock-termination. RESULTS: Midazolam (0.03-1.0 mg/kg), zolpidem (0.1 3.0 mg/kg), and zaleplon (0.1-3.0 mg/kg) increased rates of punished responding and decreased rates of nonpunished responding under the multiple schedule. Pretreatment with flumazenil (0.3-1.0 mg/kg) antagonized the anticonflict and response rate-decreasing effects of all three agonists. Pretreatment with beta CCt (3-10 mg/kg) antagonized the anticonflict and rate-decreasing effects of midazolam, as well as the rate-decreasing effects of zolpidem and zaleplon. However, beta-CCt did not antagonize the anticonflict effects of zolpidem and zaleplon; instead, these effects of zolpidem and zaleplon were apparently enhanced in the presence of beta-CCt. Under the schedule of stimulus shock termination, both flumazenil and beta-CCt antagonized zolpidem and zaleplon; however, the effects of beta-CCt were less consistent than the effects of flumazenil. CONCLUSION: In nonhuman primates, different types of GABAA receptors may mediate the anticonflict and the response rate-decreasing effects of the nonselective GABAA agonist midazolam and the selective GABAA1 agonists zolpidem and zaleplon. PMID- 11498726 TI - Tryptophan depletion increases aggression in women during the premenstrual phase. AB - RATIONALE: Reducing serotonin by the method of tryptophan depletion (TD) has led to increased aggression but experimental studies have not used female subjects. OBJECTIVE: To evaluate the effects of TD on aggression in women in the late luteal phase of their menstrual cycle. METHODS: Healthy women were recruited and randomly assigned to an amino acid drink either depleted or with a balanced amount of tryptophan. At 4.5 h later, they competed on the competitive reaction time task. RESULTS: Women who had received the TD drink showed more behavioural aggression in response to provocation. CONCLUSION: Decreased serotonergic neurotransmission increases aggression in women as well as men. PMID- 11498727 TI - The chronic effects of an extract of Bacopa monniera (Brahmi) on cognitive function in healthy human subjects. AB - RATIONALE: Extracts of Bacopa monniera have been reported to exert cognitive enhancing effects in animals. However, the effects on human cognition are inconclusive. OBJECTIVE: The current study examined the chronic effects of an extract of B. monniera (Keenmind) on cognitive function in healthy human subjects. METHODS: The study was a double-blind placebo-controlled independent group design in which subjects were randomly allocated to one of two treatment conditions, B. monniera (300 mg) or placebo. Neuropsychological testing was conducted pre-(baseline) and at 5 and 12 weeks post drug administration. RESULTS: B. monniera significantly improved speed of visual information processing measured by the IT task, learning rate and memory consolidation measured by the AVLT (P<0.05), and state anxiety (P<0.001) compared to placebo, with maximal effects evident after 12 weeks. CONCLUSIONS: These findings suggest that B. monniera may improve higher order cognitive processes that are critically dependent on the input of information from our environment such as learning and memory. PMID- 11498728 TI - Histochemical examination of osteoblastic activity in op/op mice with or without injection of recombinant M-CSF. AB - Osteopetrotic (op/op) mice do not exhibit bone remodeling because of defective osteoclast formation caused by the depletion of macrophage colony-stimulating factor (M-CSF). In the present study, we investigated tibial bones of op/op mice with or without prior injections of M-CSF to determine whether osteoclast formation and subsequent bone resorption could activate osteoblasts, which is known as a "coupling" phenomenon. In op/op mice, no osteoclasts were present, but the metaphyseal osteoblasts adjacent to the growth plate cartilage seemed to be active, revealing an intense alkaline phosphatase (ALPase) immunoreactivity. Consequently, primary trabecular bones were extended continuously to the diaphysis, indicating that bone modeling is well achieved in op/op mice. In contrast with the metaphysis, most of the diaphyseal osteoblasts were flattened and showed weak ALPase activity, and, as a result, they seemed to be less active. Osteopontin (OPN) was localized slightly at the interface between bone and cartilage matrices of the primary trabeculae. In contrast, in op/op mice injected with M-CSF, tartrate-resistant acid phosphatase-positive osteoclasts appeared, resorbing trabecular bones of the diaphyseal region. The diaphyseal osteoblasts in the vicinity of the active osteoclasts were cuboidal and exhibited strong ALPase immunoreactivity. OPN was observed not only at the bone-cartilage interface, but also significantly on the resorption lacunae beneath the bone resorbing osteoclasts. These observations indicate that the activation of diaphyseal osteoblasts appears to be coupled with osteoclast formation and subsequent osteoclastic bone resorption. Alternatively, the metaphyseal osteoblasts at the chondro-osseous junction seemed to be less affected by osteoclastic activity. PMID- 11498729 TI - 1alpha-Hydroxyvitamin D3 suppresses trabecular bone resorption by inhibiting osteoclastogenic potential in bone marrow cells after ovariectomy in mice. AB - To test the hypothesis that 1alpha-hydroxyvitamin D3 (1alpha(OH)D3) suppresses bone resorption after ovariectomy (ovx) by inhibiting osteoclastogenic potential in bone marrow cells, the bilateral tibiae of ddY mice, 8 weeks of age, subjected to ovx were obtained. 1alpha(OH)D3, at doses of 0, 0.2 (low dose), or 0.4 microg/kg body weight (high dose), was administered orally by canula three times a week for 2 or 6 weeks. Histomorphometric analysis of the proximal tibiae revealed that 1alpha(OH)D3 administration had no significant effect on trabecular bone volume of ovx limbs, which was reduced after ovx. The bone formation rate, increased by ovx, was significantly decreased by the administration of high-dose 1alpha(OH)D3. The ovx-induced increases in osteoclast number and surface at 2 weeks postsurgery were suppressed by the administration of high-dose 1alpha(OH)D3. With regard to bone marrow cells, the number of nonadherent cells per tibia obtained from ovx limbs increased, and this increase was suppressed by the administration of low- and high-dose 1alpha(OH)D3. The formation of mineralized nodules in marrow cultures obtained from ovx limbs was increased after surgery and unaltered by 1alpha(OH)D3 administration. The number of osteoclast-like multinucleated cells obtained from ovx limbs was reduced by low- and high-dose 1alpha(OH)D3 administration alike. The number of colony forming units-fibroblast and the number of colony forming units for granulocytes and macrophages was unaltered by ovx or the administration of 1alpha(OH)D3. The present study clearly demonstrates that high-dose 1alpha(OH)D3 suppresses osteoclast numbers and surface after ovx. The inhibitory effects of low- and high dose 1alpha(OH)D3 on bone marrow cells after ovx were marked in the differentiation from osteoclast precursors to mature osteoclasts. Administration of 1alpha(OH)D3 suppressed ovx-promoted trabecular bone resorption by inhibiting osteoclastogenic potential in bone marrow cells. PMID- 11498730 TI - Hypophosphatemic rickets accompanying McCune-Albright syndrome: evidence that a humoral factor causes hypophosphatemia. AB - McCune-Albright syndrome (MAS) is sometimes complicated by hypophosphatemia. However, it remains unclear whether a humoral factor is associated with the cause of hypophosphatemia. We isolated cells with mutations of the Gsalpha gene from fibrous bone dysplasia tissues of two MAS patients (MAS cells). Severe combined immunodeficiency (SCID) mice were subjected to experiments using from one of these cells patients. Effects of conditioned media (CM) isolated from MAS cells (MAS-CM) on phosphate transport were investigated by using rat renal slices, the renal cell line OK-B, rat intestinal rings and the human intestinal cell line Caco-2. In addition, the effects of MAS-CM on human sodium-dependent phosphate transporter (NPT2) gene promoter activity expression were investigated in the renal cell line OK-B2400 and were compared with the effects of CM isolated from a patient with oncogenic hypophosphatemic osteomalacia (OHO). MAS cells caused significant hypophosphatemia (P < 0.05) and elevated serum alkaline phosphatase activity (P < 0.05) in SCID mice. The MAS-CM significantly inhibited phosphate uptake in everted intestinal rings (P < 0.01), whereas it had no effect on glucose uptake. The MAS-CM had no effect on either phosphate uptake in the kidney or NPT2 gene promoter activity. In contrast, the CM of the OHO patient significantly inhibited phosphate uptake and NPT2 gene promoter activity. These results indicate that the humoral factor derived from fibrous dysplasia cells of the MAS patient is different to that from OHO patients, because the humoral factor from the MAS patient inhibited phosphate transport not in the kidney but in the intestine. PMID- 11498731 TI - A polymorphic variant at the Werner helicase (WRN) gene is associated with bone density, but not spondylosis, in postmenopausal women. AB - Werner syndrome (WS) is a rare autosomal recessive progeroid syndrome characterized by the premature onset of multiple age-related disorders. The gene responsible for WS has been identified as WRN, a member of the RecQ family of helicase genes. Based on the fact that patients with WS exhibit osteoporosis and osteoarthritis, the present study was undertaken to clarify the contribution of the WRN gene to the etiology of these two common age-related disorders in normal postmenopausal women. We investigated the association of a WRN gene polymorphism, namely c.4330 T --> C leading to an amino acid substitution from Cys to Arg, with bone density and lumbar spondylosis score in unrelated Japanese postmenopausal women (n = 377). Genotypic frequencies of T/T, T/C, and C/C were 87.5%, 12.2%, and 0.3%, respectively. Bone density of the lumbar spine (L2-4) was significantly lower in women carrying the minor C allele than in non-carriers (P = 0.037). When bone density was expressed by the Z score after being adjusted by age and weight, carriers of the C allele showed lower values not only in the lumbar spine, but also in the total body (P = 0.015 and 0.042, respectively). The association study with spondylosis in postmenopausal women (n = 221) revealed that this polymorphism was not related to the severity of spondylosis expressed by the Kellgren-Lawrence score at any disk level of the lumbar spine (L2/3-L5/S1). These findings indicate that the WRN gene may be a candidate for the genetic regulation of osteoporosis, but not spondylosis, in normal Japanese postmenopausal women. PMID- 11498732 TI - Vitamin D receptor gene polymorphism is related to bone density, circulating osteocalcin, and parathyroid hormone in healthy adolescent girls. AB - Bone mineral density (BMD) in adolescence is under strong genetic control and may influence the risk of future osteoporosis and resulting fracture. We investigated the vitamin D receptor (VDR) gene polymorphisms ApaI, BsmI, FokI, and TaqI, in relation to BMD, circulating calcium, osteocalcin, and parathyroid hormone (PTH) concentrations in healthy Caucasian girls (n = 99; mean (+/- SD) age 16.9 +/- 1.2 years). BMD of the total body, femoral neck, and lumbar spine, and bone area of the femur, lumbar spine, and total body were measured using dual energy X-ray absorptiometry. BMD values were adjusted for age, body height, body weight, and physical activity. Using ANOVA, the ApaI genotype Aa had lower circulating levels of osteocalcin (P < 0.01), higher levels of PTH (P = 0.04), and there was a strong tendency toward a significantly higher lumbar spine BMD (P = 0.08) compared with aa subjects. BMD of the lumbar spine (P = 0.02), but not femoral neck or total body, was higher in Bb subjects compared with their bb counterparts. There was no difference in BMD at any measured site of the FokI alleles. There was a strong tendency for a higher BMD at the lumbar spine of Tt subjects compared with TT subjects (P = 0.05). Neither of the different VDR polymorphisms was related to BMD before adjustment for age, body weight, body height, and physical activity. In conclusion, VDR gene polymorphism, defined by ApaI, is related to differences in circulating osteocalcin and PTH, and BsmI is related to lumbar spine BMD in healthy adolescent girls. The results stress the importance of adjusting BMD for confounding factors, such as body weight and physical activity, in order to be able to find any genotype effect on BMD. PMID- 11498733 TI - Possible involvement of vitamin D receptor gene polymorphism in male patients with ossification of spinal ligaments. AB - Ossification of spinal ligaments (OSL) is a common form of myelopathy characterized by heterotopic bone formation in the spinal ligaments, predominantly in men. Although the etiology of OSL is not fully understood, previous studies have strongly suggested the involvement of genetic factors in this disease. To investigate the possible involvement of vitamin D receptor (VDR) gene polymorphism in Japanese male patients with OSL, we analyzed: (a) the VDR genotype defined by BsmI polymorphism in patients with obvious OSL and controls; and (b) the effect of 1,25-dihydroxyvitamin D3 on alkaline phosphatase (ALP) activity of spinal ligament cells derived from patients without OSL. With regard to the VDR genotype, of the patients with OSL (n = 27), none had the BB genotype (0%), one had the Bb genotype (4%), and 26 had the bb genotype (96%). In the control group (n = 97) three had the BB genotype (3%), 18 had the Bb genotype (19%), and 76 had the bb genotype (78%). As a result, the B allele frequency in patients with OSL (2%) was significantly lower than in controls (12%). 1,25 Dihydroxyvitamin D3, at concentrations of 10-9 and 10-8 M, significantly increased ALP activity of the ligament cells (n = 8), suggesting that 1,25 dihydroxyvitamin D3 is able to promote osteogenic differentiation of normal ligament cells. Among the Japanese, sensitivity to vitamin D has been reported to vary between the alleles of the VDR; i.e., bone mineral density (BMD) in patients without the B allele is increased by vitamin D treatment, whereas patients with the B allele do not show such an increase in BMD. The present investigation is a small preliminary study, but the findings suggest, for the first time, that the B allele of the VDR acts as an inhibitor in the pathogenesis of human male OSL. PMID- 11498734 TI - Insufficiency fracture of the spine: a prospective analysis based on radiographic and scintigraphic diagnosis. AB - To investigate the clinical characteristics of insufficiency fracture of the spine, patients aged 65 years or older, who had been hospitalized for acute low back pain without either an acute traumatic event or a previous history of malignant disease, were examined by plain radiographs and technetium-99m methylene disphosphonate bone scanning. Fifteen patients were diagnosed as having a vertebral insufficiency fracture on the basis of: (a) radiographic oseteopenia of the vertebra; (b) positive bone scan images; and (c) no episode of newly identified malignant disease in a subsequent 1 year follow up. These patients were analysed with respect to age, gender, bone mineral density, the number and distribution of the affected vertebrae, the extent of vertebral deformity, and consistency between radiographic and bone scan findings. Vertebral insufficiency fractures exhibited: (a) strict female predominance; (b) concave deformity of the affected vertebra; (c) a wide range of the vertebral height ratios and fracture distribution; and (d) low consistency between the vertebral deformity assessed by the lateral radiograph and positive activity on bone scanning. These findings represent a unique clinical feature of vertebral insufficiency fracture, and also emphasize the difficulty of radiographic diagnosis, as well as the importance of a comprehensive diagnostic approach. PMID- 11498735 TI - Carpal tunnel syndrome leads to significant bone loss in metacarpal bones. AB - The present study was designed to determine bone density modifications at the forearm and metacarpal bones in patients with carpal tunnel syndrome (CTS). Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry at the one third distal end and for the total of the radius-ulna, together with the third and fourth metacarpal bones, in 48 clinically and electrophysiologically diagnosed (18 unilateral and 15 bilateral) affected extremities in 33 premenopausal women (mean age 38.9 +/- 6.5 years) with CTS. BMD values for non affected extremities were used as controls for comparison with affected extremities. Bone mass was decreased approximately 7% in the forearm region (P < 0.02) and 18% in metacarpal bones (P < 0.01) of the thenar atrophy associated group compared with controls. A significant correlation was observed between disease duration (mean duration 3.2 +/- 2.7 years) and the decrease in metacarpal bone density (r = 0.43; P = 0.004). This is the first clinical report of quantified bone loss in affected extremities in patients with CTS, and the results suggest the need for further studies to assess the clinical significance and morbidity of this pathology, especially in patients with thenar atrophy. PMID- 11498736 TI - Studies on the frequencies of vitamin D receptor gene polymorphism in postmenopausal women of Han and Kazak nationality in China. AB - Polymorphyism of the vitamin D receptor (VDR) gene is reported to play a major role in variations of the genetic regulation of bone mass. The role of VDR polymorphisms within various ethnic populations is also undetermined. The purpose of the present study was to determine the frequencies of VDR genotypes in the Han and Kazak nationalities in China. We analyzed the polymorphism defined by the Bsm1 restriction enzyme using polymerase chain reaction-restriction fragment length polymorphism in 179 healthy postmenopausal Han women and 63 healthy postmenopausal Kazak women. The genotype frequencies of the VDR were calculated later using the Hardy-Weinberg equilibrium formula. The results showed that for women of Han nationality, the bb, Bb, and BB genotypes accounted for 90.5%, 9.5%, and 0%, respectively. In Kazak women, the respective frequencies were 38.1%, 55.56%, and 6.35%. We found that there was a significant difference between women of Han and Kazak nationality in terms of the frequency of distribution of VDR genotype (P < 0.001). There was a similar distribution of VDR genotypes in Kazak women to that seen for the Caucasian population of the USA. The results of the present study provide further evidence on the different pathogeny of osteoporosis in various ethnic groups. PMID- 11498737 TI - Distribution of bone mineral density in the proximal tibia in mid-teens. AB - The distribution of bone mineral density (BMD) in the proximal tibia was studied in 94 knees from 47 mid-teens. The subjects were healthy high school freshmen (19 male; 28 female) who engaged in daily physical exercise. We measured BMD at six regions of interest (ROI) at the bilateral proximal tibia (three immediately above and three immediately be-low the physeal line) using dual-energy X-ray absorptiometry (QDR-2000; Hologic, Waltham, MA, USA). Subjects were supine during the measurements, with the knee extended and the foot fixed in a special holding device, ensuring correct rotational alignment of the knee. Measurements were performed using the forearm array scan mode. The precision of this method (coefficient of variation) was 1.1%-4.9% (mean 2.5%) for six ROIs. The BMD of the medial side was higher than that of the lateral side, and the BMD at the center of the metaphysis was higher than that of the epiphysis. The subjects in the present study did not show any misalignment of the knee joints. These findings indicate that the medial side of the proximal tibia already has higher BMD than the lateral side in teenagers with normal alignment of the knee joint, and that the increase in epiphyseal BMD may be slower than that of metaphyseal BMD. It is suggested that this phenomenon should be life-long and may play a role in the progression of osteoarthritis of the knee. PMID- 11498739 TI - Kinetics of standardized large volume leukapheresis (LVL) in patients do not show a recruitment phenomenon of peripheral blood progenitor cells (PBPC). AB - Although several studies have demonstrated the efficacy of large volume leukapheresis (LVL) to yield high numbers of peripheral blood progenitor cells (PBPC), the mechanisms of stem cell release into circulation and the postulated phenomenon of PBPC recruitment during apheresis have not been investigated in detail. Therefore, we analyzed the kinetics of stem cell enrichment in a total of 34 standardized LVL for patients with hematologic malignancies (lymphoma, multiple myeloma) and solid tumors (breast cancer, rhabdomyosarcoma). LVL was started 2 h after administration of G-CSF processing six times the patient's blood volume. Cells were sequentially collected into six bags and the numbers of leukocytes, mononuclear cells (MNC), CD34+ cells and colony-forming cells (CFU GM) in each collection bag were analyzed. The numbers of PBPC collected demonstrated a continuous decrease starting after an early maximum during the second processed blood volume (P = 0.001). Interestingly, these kinetics of decreasing stem cell yields during LVL were similar for both entities of patients with hematologic malignancies as well as for both groups of patients with solid tumors. In summary, a recruitment phenomenon, defined as a time-dependent and LVL induced increase of PBPC, could not be demonstrated in any of the diseases investigated. PMID- 11498738 TI - Collection of hematopoietic stem cells from patients with autoimmune diseases. AB - We reviewed data from 24 transplant centers in Asia, Australia, Europe, and North America to determine the outcomes of stem cell collection including methods used, cell yields, effects on disease activity, and complications in patients with autoimmune diseases. Twenty-one unprimed bone marrow harvests and 174 peripheral blood stem cell mobilizations were performed on 187 patients. Disease indications were multiple sclerosis (76 patients), rheumatoid arthritis (37 patients), scleroderma (26 patients), systemic lupus erythematosus (19 patients), juvenile chronic arthritis (13 patients), idiopathic autoimmune thrombocytopenia (8 patients), Behcet's disease (3 patients), undifferentiated vasculitis (3 patients), polychondritis (1 patient) and polymyositis (1 patient). Bone marrow harvests were used in the Peoples Republic of China and preferred worldwide for children. PBSC mobilization was the preferred technique for adult stem cell collection in America, Australia, and Europe. Methods of PBSC mobilization included G-CSF (5, 10, or 16 microg/kg/day) or cyclophosphamide (2 or 4 g/m2) with either G-CSF (5 or 10 microg/kg/day) or GM-CSF (5 microg/kg/day). Bone marrow harvests were without complications and did not affect disease activity. A combination of cyclophosphamide and G-CSF was more likely to ameliorate disease activity than G-CSF alone (P < 0.001). g-csf alone was more likely to cause disease exacerbation than the combination of cyclophosphamide and g-csf (P = 0.003). Three patients died as a result of cyclophosphamide-based stem cell collection (2.6% of patients mobilized with cyclophosphamide). When corrected for patient weight and apheresis volume, progenitor cell yields tended to vary by underlying disease, prior medication history and mobilization regimen. Trends in the approaches to, and results of, progenitor cell mobilization are suggested by this survey. While cytokine-based mobilization appears less toxic, it is more likely to result in disease reactivation. Optimization with regard to cell yields and safety are likely to be disease-specific and prospective disease-specific studies of mobilization procedures appear warranted. PMID- 11498740 TI - Allogeneic haemopoietic cell transplants in Australia, 1996--a multi-centre retrospective comparison of the use of peripheral blood stem cells with bone marrow. AB - A retrospective comparison was carried out on adult patients receiving HLA identical allogeneic haemopoietic stem cell transplants from siblings in Australia in 1996, comparing bone marrow with G-CSF-mobilised peripheral blood stem cells. A total of 131 transplant recipients from nine centres were included in this study, of whom 79 received bone marrow, 44 blood stem cells and eight both. All but three of the 131 patients had cyclosporin and methotrexate as graft versus-host disease prophylaxis. The minimum follow-up time for surviving patients is 27 months. Comparisons were carried out between the BM and PBSC groups. There were no significant differences between groups in age, sex, diagnosis, donor characteristics or pretransplant conditioning. Median time to neutrophil recovery of 0.5 x 10(9)/l was 14 days for PBSC recipients, compared to 19 days for marrow recipients (P < 0.0005). median time to platelet recovery of 20 x 10(9)/l was 17 days for PBSC recipients, compared to 28 days for marrow recipients (P < 0.0005). there were no significantly increased risks of either acute or chronic GVHD in the PBSC recipients. there were no significant differences between the groups in the incidence of major transplant-related complications, disease-free survival or overall survival. PMID- 11498741 TI - Comparison of new patients with Bence-Jones, IgG and IgA myeloma receiving sequential therapy: the need to regard these immunologic subtypes as separate disease entities with specific prognostic criteria. AB - Of the 61 newly diagnosed patients with Bence-Jones (BJ) myeloma presenting to our centre between May 1986 and December 1997, 53 received sequential therapy (ST) comprising infusional chemotherapy (IC) followed by high-dose therapy and autotransplantation with interferon-alpha2b maintenance. The outcome was compared with 153 IgG and 39 IgA similarly treated myeloma patients. Response to IC and high-dose was comparable between the three subtypes but a significantly higher proportion of patients with BJ myeloma failed to receive high-dose compared to IgG (P = 0.003) and IgA (P = 0.04) myeloma. Median overall survival (OS) of patients with BJ myeloma (2.8 years) and event-free survival (EFS, 1.2 years) was significantly shorter than for patients with IgG myeloma (4.5 years, P = 0.03 and 2.1 years, P = 0.03, respectively). However, among those patients who achieved complete remission there was no difference in OS and EFS between IgG and BJ myeloma. In distinction to IgG myeloma where age and beta2M were significant, Cox analysis on presentation features identified performance status and urine total protein as having significant impact on OS. We conclude that achieving CR is an important treatment aim in patients with BJ myeloma, conferring a similar outlook on survival as in patients with the IgG subtype, and there is a need to consider different subtype-specific staging systems when evaluating the results of published or ongoing therapeutic trials. PMID- 11498742 TI - Do corticosteroids add any benefit to standard GVHD prophylaxis in allogeneic BMT? AB - In a retrospective study, we compared 15 patients who received cyclosporine (CsA), methotrexate (MTX) and prednisone (PDN) and 15 patients who received CsA MTX for GVHD prophylaxis after allogeneic BMT (HLA-identical sibling (n = 22), related one HLA mismatch (n = 1), unrelated matched donors (n = 6), unrelated one HLA mismatch (n = 1)). The primary objectives of this study were to compare the incidence of GVHD and post-transplantation complications. Secondary objectives were to compare relapse rate, transplant-related mortality and overall survival. The incidence of acute GVHD grade III-IV was similar between the two groups (P = 0.66), as was the incidence of chronic GVHD (P = 0.67). Incidence of arterial hypertension was significantly higher in patients who received prophylactic PDN, (P = 0.03) and more insulin treatment was required in this group (P = 0.003). We observed no differences in the incidence of infections or upper digestive tract bleeding. Musculoskeletal complications appeared earlier in the group which received PDN. With a median follow-up of 4.4 years, patients in the CsA-MTX group had better overall survival, 46.7% vs 13.3% (P = 0.026). Relapse was a more frequent cause of death in the CsA-MTX group, whereas procedure-related mortality was more frequent in the CsA-MTX-PDN group (P = 0.013). These results suggest that prophylactic prednisone when combined with cyclosporine and methotrexate adds no benefit in acute or chronic GVHD prevention and may increase the morbidity of allogeneic transplantation. Corticosteroids may be reserved for GVHD treatment. PMID- 11498743 TI - Treatment of severe steroid refractory acute graft-versus-host disease with infliximab, a chimeric human/mouse antiTNFalpha antibody. AB - Acute graft-versus-host disease (aGVHD) is a serious complication of allogeneic peripheral blood stem cell transplantation (PBSCT). Patients with severe aGVHD not responding to treatment with steroids have a poor prognosis. We treated four patients with severe aGVHD refractory to steroids with infliximab, a chimeric human/mouse antiTNFalpha antibody. Patients (CML 2, MM 1, AML 1) developed grade III-IV GVHD at a median of 34 days (range 15-76) after myeloablative PBSCT (two), donor lymphocyte infusion for relapsed CML (one) or non-myeloablative PBSCT (one), respectively. All patients had severe intestinal involvement in addition to skin and/or liver disease and had received treatment with high-dose steroids (four) for a median of 11 days (range 5-17) in addition to CsA (four) and MMF (three). Infliximab (10 mg/kg) was given once a week until clinical improvement. In three of four patients a complete resolution of diarrhea and significant improvement of skin and liver disease were observed. Two patients received one, one patient two and one patient three infliximab infusions. At present two patients are alive >200 days after therapy, one with limited cGVHD. Two patients died, one of progressive malignant disease without GVHD and one of refractory GVHD. Infliximab is apparently an active drug for the treatment of aGVHD. PMID- 11498744 TI - Adenoviral infection after allogeneic stem cell transplantation (SCT): report on 130 patients from a single SCT unit involved in a prospective multi center surveillance study. AB - The incidence of adenovirus (AV) infections following SCT was determined in a prospective multicenter trial. Over 1 year, 130 consecutive patients undergoing allogeneic SCT at Essen University Hospital were included and followed for 6 months. Source of stem cells was blood in 68 cases. Fifty-eight patients had HLA identical sibling donors. Throat swabs, urine and stool samples were screened weekly for AV antigen and DNA by ELISA and nested PCR, respectively. In 35 cases adenovirus infection was detected. There was no seasonal variation. Throat swabs were positive in 24, urine in 12, and stool in 11 cases, resulting in a cumulative risk of infection of 29%. The incidences of AV infection of the respiratory, gastrointestinal and urinary tract were 19%, 10%, and 9%, respectively, and infections were diagnosed after a median (range) interval of 44 (-2-179), 37 (-2-168), and 53 (17-153) days after transplantation. On multivariate analysis, presence of AV antibody in the donor and acute graft versus-host disease grade IV were found to be independent risk factors for AV infection. Eleven patients had AV isolated from more than one site and five patients had probable AV disease. We were not able to identify patients in whom AV infection was the leading cause of death. The majority of patients infected with AV suffered from severe acute graft-versus-host disease often accompanied by other opportunistic infections, such as aspergillosis or CMV reactivation. Nineteen out of 36 patients who died during the observation period had AV infection. In summary, AV infection after allogeneic SCT was observed in a substantial number of patients. In addition to well-known risk factors for viral infection after SCT we were able to demonstrate that a positive AV antibody test in the donor is an important risk factor for AV infection. Further studies are needed, however, before final conclusions on the clinical sequelae of AV infection can be made and the role of preventive and therapeutic strategies toward AV infection after allogeneic SCT can be defined. PMID- 11498745 TI - Gastroparesis following bone marrow transplantation. AB - Patients often develop nausea, vomiting and bloating after bone marrow transplantation (BMT). These symptoms may interfere with nutrition and the ability to take oral medications. Gastroparesis is a recognized cause of these symptoms in non-transplant patients but less is known about patients who undergo BMT. Between January 1996 and March 1997, a total of 151 patients underwent BMT. Eighteen patients (12%) developed persistent symptoms suggestive of gastroparesis (persistent nausea, vomiting or bloating). Scintigraphic gastric emptying studies were performed to assess for gastroparesis. Prokinetic agents were administered at the time of study. The records on these patients were compared with those of all other patients undergoing BMT during the same time period without these symptoms. Nine patients who demonstrated delayed gastric emptying were further evaluated with esophagastroduodenoscopy and biopsy. Biopsy samples were reviewed for evidence of graft-versus-host disease (GVHD). Fourteen of 18 patients demonstrated delayed gastric emptying and most responded to prokinetic agents given at the time of study. Age, conditioning regimen, cytomegalovirus antigenemia and acute GVHD did not appear to be associated with the development of gastroparesis. Allogeneic BMT recipients were at higher risk than autologous BMT patients (26% vs 0%, P < 0.0001). of allogeneic bmt recipients, there was a nonsignificant trend of patients receiving tacrolimus to be less likely to experience gastroparesis than those receiving cyclosporine (27% vs 48%, P = 0.08). For the nine patients undergoing upper endoscopy, GVHD on gastric biopsy was an uncommon finding and was mild when present. Gastroparesis appears to be a common cause of nausea, vomiting and bloating following allogeneic BMT. This may occur less often with tacrolimus than cyclosporine because of the former agent's prokinetic properties. Patients usually respond to prokinetic drugs at the time of scintigraphy. GVHD and CMV infection do not appear to be major contributing factors. PMID- 11498746 TI - Changes in autoimmune thyroid disease following allogeneic bone marrow transplantation. AB - Autoimmune diseases can be transmitted and eliminated by bone marrow transplantation (BMT). There have been several cases of autoimmune thyroid disease (AITD) occurring after BMT, but AITD remission has been rarely reported. We present four cases in which the remission or transfer of AITD occurred after an allogeneic BMT. Two patients with severe aplastic anemia (SAA) showed evidence of remission of Hashimoto's thyroiditis which they had before allogeneic BMT. One patient with SAA, which developed during treatment with propylthiouracil for Graves' disease, underwent allogeneic BMT and showed evidence of Graves' disease remission following BMT. In one patient, new AITD occurred after an allogeneic BMT from an HLA-matched sibling who already had AITD. These cases support the evidence that the immune system is newly reconstituted after BMT, and severe autoimmune disease can be an indication for BMT. To fully understand the real changes in autoimmune status after BMT, long-term prospective studies are necessary. PMID- 11498747 TI - Factors affecting gonadal function after bone marrow transplantation during childhood. AB - Conditioning for bone marrow transplantation (BMT) may alter viability of germ cells and production of gonadal hormones. We analyzed the risk factors for gonadal failure after 12 Gy total body irradiation (TBI) given as six fractions (n = 31, group 1), 10 Gy (one dose) TBI (n = 20, group 2), 6 Gy (one dose) total lymphoid irradiation (TLI, n = 17, group 3) and chemotherapy alone (n = 7, group 4), given at 7.7 +/- 0.4 (0.6-13.6) years. Among the 34 girls, seven (20.6%) had normal ovarian function with regular spontaneous menstruation and normal plasma follicle-stimulating (FSH) and luteinizing (LH) hormones, five (14.7%) had partial ovarian failure with regular menstruation but increased FSH and/or LH, and 22 (64.7%) had complete ovarian failure. The 24 girls with chronological and bone ages >13 years included similar percentages, with increased FSH or LH in all four groups. There was a positive correlation between age at BMT and FSH (r = 0.54, P < 0.01), but not with lh, and between fsh and lh (r = 0.8, P = 0.0003). Plasma FSH concentrations had returned to normal spontaneously in six cases, and those of LH in two cases. Among the 41 boys, 16 (39%) had normal testicular function and 25 (61%) had tubular failure and increased FSH. Of these, 10 also had Leydig cell failure (three complete and seven partial). The 18 boys with chronological and bone ages >15 years included similar percentages with increased FSH or LH in groups 1 to 3, and testicular volume was significantly lower in group 2 than in group 3 (P = 0.008). There was no correlation between age at BMT and FSH, LH or testosterone, but there was a negative correlation between FSH and inhibin B (rho = -0.87, P < 0.003). we conclude that girls are more likely to suffer ovarian failure the older they are at bmt, and that early ovarian recovery is possible. the negative correlation between fsh and inhibin b in boys suggests that this parameter is an additional indicator of tubular function. PMID- 11498748 TI - Correlation between HHV-6 infection and skin rash after allogeneic bone marrow transplantation. AB - We investigated whether a causal relationship exists between human herpesvirus 6 (HHV-6) and skin rash resembling acute graft-versus-host disease (GVHD) following bone marrow transplantation (BMT). Isolation of HHV-6 was used to monitor active HHV-6 infection in this study. We analyzed 25 episodes of skin rash in 22 recipients. All recipients were seropositive for HHV-6 before BMT. The onset of skin rash started prior to 30 days post transplantation (group A) in 15 of 25 cases, but after that (group B) in the remaining 10 cases. Twenty-five skin tissue samples were obtained from 22 recipients. The HHV-6 genome was detected in four of 15 skin samples from group A, but not detected in those from group B. HHV 6 was isolated from 11 of 22 recipients around 2 to 3 weeks after BMT (range 14 to 28 days after BMT). HHV-6 was isolated at a time between 10 days before and after the onset of skin rash (skin rash-related viremia) in nine cases in group A. Meanwhile, no skin rash-related viremia was observed in group B. Of the four recipients with positive detection of HHV-6 genome in their skin tissue (group A), two had HHV-6 viremia at the same time. The association between the timing of HHV-6 infection and the onset of skin rash was analyzed statistically. HHV-6 viremia (skin rash-related viremia) was found in nine of 15 (60%) cases in group A, compared with none of 10 (0%) cases in group B. This difference was statistically significant (P = 0.008). Moreover, HHV-6 infection (skin rash related viremia and/or positive detection of HHV-6 DNA in skin tissue) was demonstrated in 11 of 15 (73.3%) cases in group A, compared with none of 10 (0%) cases in group B (P = 0.001). Thus, this study suggests that HHV-6 may be involved in the development of skin rash in the first month after allogeneic BMT. PMID- 11498749 TI - Successful unrelated bone marrow transplantation for a patient with chronic granulomatous disease and associated resistant pneumonitis and Aspergillus osteomyelitis. AB - We describe the successful treatment of a 20-year-old patient with chronic granulomatous disease (CGD), by unrelated bone marrow transplantation (UBMT). The patient is relatively old compared to other CGD patients treated with BMT. He had had repeated serious infections from early childhood and was diagnosed as CGD, gp91-phox deficiency. Prolonged antibiotic-resistant pneumonitis worsened when the patient was 18 years old. In addition, he suffered Aspergillus osteomyelitis and acute renal failure due to amphotericin B. He received 94 granulocyte transfusions from 94 adult donors and the infections gradually improved. In September 1998, at 20 years of age, he underwent UBMT from an HLA 6 antigen matched male donor, with CY and TBI conditioning. He received MTX and CsA as prophylaxis against GVHD. No serious complications occurred and rapid engraftment was achieved. Acute GVHD (grade 2, at day 19) and chronic GVHD (limited, at day 192) occurred. However, both were easily controlled. The patient is alive and well with no late rejection 26 months after UBMT. PMID- 11498750 TI - Acute autoimmune hemolytic anemia following unrelated cord blood transplantation as an early manifestation of chronic graft-versus-host disease. AB - A 16-month-old girl diagnosed with osteopetrosis underwent an unrelated, partially matched (with major mismatch at A locus) cord blood stem cell transplant. Twelve months later she developed severe acute autoimmune hemolytic anemia (AIHA). Immunophenotype analysis of lymphocyte subsets 8 months post transplant showed a low number of T lymphocytes, with normal subsets, and with NK cells and B lymphocytes within normal ranges. When the hemolytic anemia developed, the lymphocytes subsets changed and analysis showed higher numbers of B lymphocytes than previously, lower CD3+ T lymphocytes with inversion of the CD4/CD8 ratio and an abnormal proportion of T lymphocyte subsets. She was being treated with cyclosporine, and steroids and immunoglobulins were added. Initially the AIHA improved, but repeated infectious episodes led us to tail off the immunosuppressive treatment. The AIHA relapsed and cyclosporine was restarted. Currently, she is on cyclosporine and low-dose steroid treatment with no hemolytic features. During the 3 months when the AIHA was being treated, she developed extensive skin cGVHD and recurrent pneumothoraces. AIHA may be the first manifestation of abnormal reconstitution of immunity developing after a hematopoietic transplant. This abnormal reconstitution is also the basis of cGVHD. We suggest that aggressive immunosuppressive treatment with intensive measures against infection could give a better prognosis to such patients. PMID- 11498751 TI - Successful HLA-identical bone marrow transplantation in a patient with PNP deficiency using busulfan and fludarabine for conditioning. AB - PNP deficiency is an autosomal recessive metabolic disorder characterized by severe combined immunodeficiency and by complex neurological symptomatology including ataxia, developmental delay and spasticity. Patients usually die in the first or second decade of life due to recurrent infections. The only curative treatment is bone marrow transplantation (BMT). We describe a 22-month-old girl who underwent BMT from her HLA-identical brother. Conditioning consisted of busulfan and fludarabine only, resulting in low toxicity and prompt engraftment. At 18 months after BMT, the girl has developed normal immunological functions, and her neurological status has improved. PMID- 11498752 TI - Beneficial effect of intravenous lidocaine in cutaneous chronic graft-versus-host disease secondary to donor lymphocyte infusion. AB - We report two cases of refractory chronic graft-versus-host disease after donor lymphocyte infusions in which the skin lesions improved dramatically with the use of intravenous pulses of lidocaine. This form of therapy has been used successfully for the cutaneous involvement of scleroderma and may have vasodilator and anti-inflammatory effects. PMID- 11498753 TI - Acute left ventricular failure following melphalan and fludarabine conditioning. AB - Cardiotoxicity has rarely been reported as a complication of melphalan or fludarabine administration as single agents. Recently, melphalan and fludarabine have been used in combination as non-myeloablative conditioning chemotherapy prior to allogeneic stem cell transplantation. We have observed the development of severe left ventricular failure in three of 21 patients treated with this combination. Cardiotoxicity in this context has not previously been reported and has implications for the assessment, monitoring and treatment of patients undergoing pre-transplant conditioning with melphalan and fludarabine. PMID- 11498754 TI - Successful mismatched sibling cord blood transplant in Hb Bart's disease. AB - A 20-month-old girl with Hb Bart's disease, who had survived neonatal complications, underwent HLA-DR antigen mismatched sibling cord blood transplantation successfully. Immune thrombocytopenia, which occurred around 2.5 months after transplant, responded to intravenous gamma-globulin. The fetal hemoglobin level rose to a peak of 52.3% on day +69 post transplant and declined gradually during the following year. Ten percent of hemoglobin Bart's was detected 2 months after transplant and this reflects the alpha-thalassemia trait of the donor. PMID- 11498755 TI - PBSC autotransplant for inflammatory bowel disease (IBD): a case of ulcerative colitis. PMID- 11498756 TI - Unrelated cord blood transplantation in a Fanconi anemia patient using fludarabine-based conditioning. PMID- 11498757 TI - Pulmonary embolism after local urokinase instillation in a Hickman catheter. PMID- 11498759 TI - Replicating adenoviral vector-mediated transfer of a heat-inducible double suicide gene for gene therapy. AB - Tumor cells that express a fusion gene of Escherichia coli cytosine deaminase (CD) and herpes simplex virus type 1 thymidine kinase (TK) sequences activate and are subsequently killed by the nontoxic prodrugs 5-fluorocytosine and ganciclovir. We have previously developed a recombinant adenovirus containing the CD-TK fusion gene controlled by the human inducible heat shock protein 70 promoter so that heat at 41 degrees C for 1 hour induces therapeutic gene expression. This adenovirus effectively transduces heat-inducible expression of the CD-TK gene into human prostate carcinoma cells. However, because a limited number of cells in a tumor can actually be infected, we created a replicating adenoviral vector to increase CD-TK gene expression. This vector is a replication competent, E1B-attenuated adenoviral vector containing the hsp70 promoter-driven CD-TK gene (Ad.E1A(+)HS-CDTK). When human prostate adenocarcinoma DU-145 cells (mutant p53) were infected with the virus at a multiplicity of infection (MOI) of 1 or 10, the viral replication was detected within 2 days at both MOIs. Similar results were observed in human colorectal carcinoma CX-1 cells. When DU-145 cells were infected with the virus at an MOI of 10, incubated for 24 hours, heated at 41 degrees C for 4 hours, and then harvested 20 hours later, Western blot analysis demonstrated that this virus successfully produced viral E1A proteins and heat shock stimulated the CD-TK gene expression by 12.3-fold. In addition, Ad.E1A(+)HS-CDTK effectively suppressed cell proliferation by viral cytopathic effect). Unlike with a replication-incompetent virus (Ad.HS-CDTK), the cytopathic effect of the virus and cytotoxicity in the presence of the prodrugs were still observed even at low MOI (MOI=1.0). PMID- 11498760 TI - Lipid-coated polyplexes for targeted gene delivery to ovarian carcinoma cells. AB - A nonviral gene delivery vector has been developed in our laboratory based on the cationic polymer, poly(2-(dimethylethylamino)ethyl methacrylate) (p(DMAEMA)). p(DMAEMA)-based polyplexes have been successfully used for the transfection of OVCAR-3 cells in vitro. However, these polyplexes were unable to transfect OVCAR 3 cells growing in the peritoneal cavity of nude mice after intraperitoneal administration, which could be ascribed to inactivation by components (including hyaluronic acid) present in the tumor ascitic fluid. The present work aimed at (a) protecting p(DMAEMA)-based polyplexes against destabilization or inactivation by polyanions such as hyaluronic acid present in tumor ascitic fluid and (b) enhancing cellular uptake of the protected p(DMAEMA)-based polyplexes by targeting with antibody Fab' fragments. To fulfill these requirements, we have developed a detergent removal method to coat polyplexes with anionic lipids. With this method, spherical particles of approximately 125 nm, which were protected from destabilization by polyanions, were obtained. More importantly, the transfection efficiency of lipopolyplexes was unaffected in the presence of hyaluronic acid, indicating that lipid coating of polyplexes protects against destabilization by hyaluronic acid. By conjugating antibody Fab' fragments directed against the epithelial glycoprotein-2 to the lipidic surface of these lipopolyplexes, target cell-specific transfection of OVCAR-3 cells could be obtained in vitro. PMID- 11498761 TI - Efficacy of the bystander effect in the herpes simplex virus thymidine kinase mediated gene therapy is influenced by the expression of connexin43 in the target cells. AB - Tumoricidal "bystander effect" observed in the herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) gene therapy was studied between different rat glioma cell lines (9L and C6 cells) under both in vitro and in vivo conditions. For that purpose, mixed populations of wild-type cells (9Lwt and C6wt) and respective HSVtk gene-transduced cells (9Ltk and C6tk) were examined for their sensitivity to GCV. A potent in vitro bystander effect was observed in 9Lwt/9Ltk and 9Lwt/C6tk combinations but not in C6wt/9Ltk and C6wt/C6tk combinations. In vivo bystander effect studied in a subcutaneous tumor model in athymic nude mice was also potent in 9Lwt/9Ltk and 9Lwt/C6tk combinations. Because the expression of connexin43, a major protein in the connexin family gene products, in 9L cells is much higher than that in C6 cells, the results suggest that the amount of connexin in target (wild-type) cells but not in effector (HSVtk gene-bearing) cells is important for the generation of the bystander effect. This hypothesis was further confirmed by the observation that in vitro bystander effect in C6wt/C6tk combination was potentiated by transduction of the connexin43 gene to the target cells. PMID- 11498762 TI - Induction of antitumor immunity by transduction of CD40 ligand gene and interferon-gamma gene into lung cancer. AB - CD40-CD40 ligand (CD40L) interaction is an important costimulatory signaling pathway in the crosstalk between T cells and antigen-presenting cells. This receptor-ligand system is known to be essential in eliciting strong cellular immunity. Here we demonstrate that murine lung cancer cells (3LLSA) transduced with the CD40L gene (3LLSA-CD40L) were rejected in syngeneic C57BL/6 mice, but grew in CD40-deficient mice to the same extent as control tumor cells. Immunohistochemical study showed that inflammatory cells, including CD4+, CD8+ T cells and NK cells, infiltrated into the inoculated 3LLSA-CD40L tumor tissue. Inoculation of 3LLSA-CD40L cells into mice resulted in the induction of 3LLSA specific cytotoxic T-cell immunity, and the growth of parental 3LLSA tumors was inhibited when 3LLSA cells were inoculated into C57BL/6 mice mixed with 3LLSA CD40L cells or when they were rechallenged 4 weeks after 3LLSA-CD40L cells were rejected. Furthermore, co-inoculation of interferon (IFN)-gamma-transduced cells (3LLSA-IFNgamma) with 3LLSA-CD40L cells enhanced the antitumor immunity efficiently in vivo. These results indicate that the in vivo priming with CD40L- and IFN-gamma gene-transduced lung cancer cells is a promising strategy for inducing antitumor immunity in the treatment of lung cancer. PMID- 11498763 TI - Effects of adenoviral wild-type p53 gene transfer in p53-mutated lymphoma cells. AB - The present study assessed the role of adenoviral vector-mediated wild-type p53 gene transfer in B lymphoma cells. Deficiency of p53-mediated cell death is common in human cancer contributing to both tumorigenesis and chemoresistance. Lymphoma cells are being considered as suitable targets for gene therapy protocols. Recently, we reported an adenoviral protocol leading to highly efficient gene transfer to B lymphoma cells. All lymphoma cell lines (n=5) tested here showed mutations in the p53 gene locus. The aim of this work was to transduce lymphoma cells with the wild-type p53 gene. Using this protocol, 88% of Raji, 75% of Daudi, and 45% of OCI-Ly8-LAM53 cells were transfected with the reporter gene green fluorescent protein at a multiplicity of infection of 200. The expression of green fluorescent protein in CA46 and BL41 cells was 27% and 42%, respectively. At this multiplicity of infection, growth characteristics of lymphoma cell lines were not changed significantly. In contrast, cells transduced with wild-type p53 gene showed an inhibition of proliferation as well as an increase in apoptosis. Cell loss by apoptosis after p53 gene transfer was up to 40% as compared to transduction with an irrelevant vector. In addition, we determined the effects of DNA damage produced by the DNA topoisomerase II inhibitor etoposide on wild-type p53 transfected lymphoma cells. In Ad-p53 transfected Raji cells, treatment with the drug resulted in a marked increase of cell loss in comparison to Ad-beta-Gal-transfected cells (45% vs. 77%). Interestingly, performing cytotoxicity studies, we could show an increased sensitivity of Raji and Daudi cells against immunological effector cells. In conclusion, transduction of wild-type p53 into lymphoma cells expressing mutated p53 was efficient and led to inhibition of proliferation and increase in apoptotic rate in some cell lines dependent on p53 mutation. This protocol should have an impact on the use of lymphoma cells in cancer gene therapy protocols. PMID- 11498764 TI - The FBMD-1 stroma cell line secretes a unique moiety which can increase retroviral transduction of lineage-committed and primitive human peripheral blood progenitor cells. AB - Peripheral blood progenitor cells are a prime target for gene therapy approaches. As recent data point to the relevance of soluble stroma factors for the efficient transduction of progenitor cells, we tested the stroma-conditioned medium (SCM) of the two cell lines FBMD-1 and L88/5 as well as desulfated and O-sulfated heparin (HS dS and HS OS) for their effect on transduction of peripheral blood progenitor cells. We transduced CD34+ cells of nine tumor patients with the retroviral SF-MDR vector containing the human multidrug resistance 1 (MDR1) gene under serum-free conditions on the fibronectin fragment CH-296 with or without SCM. Provirus-specific polymerase chain reaction showed a median 1.6-fold higher integration rate of the transgene into committed progenitor cells for the group with added FBMD-1 SCM (P=.008). This was maintained after 2 (P=.02) and, as a trend, after 5 weeks of stroma-dependent long-term culture. We found a median 1.5 fold increase in rhodamine-123 (Rh-123) exclusion in myeloid lineage-committed progeny cells following transduction in the presence of FBMD-1 SCM (P=.0004). After 2 or 5 weeks of long-term culture, a significantly higher proportion of Rh 123(dull) cells could still be detected in the FBMD-1 SCM transduction group (P=.003 and P=.04, respectively). L88/5 SCM or HS OS or HS dS was not effective as supplement for improving gene transfer. The FBMD-1 stroma cell line appears to secrete a unique moiety, which can increase retroviral transduction of lineage committed and primitive progenitor cells. The FBMD-1 stroma activity is not attributable to heparan sulfate. PMID- 11498765 TI - Modulation of cyclophosphamide-based cytochrome P450 gene therapy using liver P450 inhibitors. AB - The sensitivity of tumors to cyclophosphamide (CPA) and other anticancer prodrugs can be substantially enhanced by transduction of tumors with a prodrug-activating mammalian cytochrome P450 (CYP) enzyme in combination with the flavoenzyme P450 reductase. This gene therapy strategy provides for intratumoral prodrug activation, but is also associated with a high level of hepatic prodrug activation, which reduces the extent of intratumoral prodrug activation and contributes to systemic drug toxicity. To address this issue, five P450 inhibitors were tested for their ability to block liver CYP2C-catalyzed CPA activation selectively, i.e., without inhibiting the corresponding intratumoral activation of CPA catalyzed by a transduced CYP2B enzyme. In vitro studies revealed that the P450 inhibitors 1-aminobenzotriazole and DDEP were preferentially inhibitory toward CYP2C-dependent liver microsomal CPA activation, whereas the P450 inhibitor SKF-525A inhibited CYP2C- and CYP2B-dependent CPA activation without P450 form selectivity. By contrast, the P450 inhibitors chloramphenicol and metyrapone preferentially inhibited CYP2B-dependent CPA activation. Rat pharmacokinetic studies confirmed the inhibitory action of these compounds in vivo, with up to a 4-fold decrease in C(max) and a 7-fold increase in apparent half-life of the activated CPA metabolite, 4-hydroxy-CPA, seen in the case of 1-aminobenzotriazole. Although the rate of hepatic CPA activation could thus be decreased substantially by P450 inhibitor treatment, the net extent of hepatic CPA activation was only modestly decreased, as judged by plasma area under-the-curve values for 4-hydroxy-CPA. Moreover, P450 inhibitor treatment did not decrease CPA's host toxicity and did not enhance the tumor growth delay response to CPA in rats bearing CYP2B1-transduced gliosarcomas. These findings are discussed in the context of P450-based gene therapy strategies and ongoing efforts to enhance anticancer drug activity by increasing the exposure of P450 expressing tumors to the P450-activated prodrug CPA. PMID- 11498766 TI - Encapsulation of c-myc antisense oligodeoxynucleotides in lipid particles improves antitumoral efficacy in vivo in a human melanoma line. AB - Phosphorothioate c-myc antisense oligodeoxynucleotides [S]ODNs (free INX-6295) were encapsulated in a new liposome formulation and the antitumor activity was compared to the unencapsulated antisense in a human melanoma xenograft. The systemic administration of INX-6295 encapsulated in stabilized antisense lipid particles (SALP INX-6295) improved plasma AUC (area under the plasma concentration-time curve) and initial half-life of free INX-6295, resulting in a significant enhancement in tumor accumulation and improvement in tumor distribution of antisense oligodeoxynucleotides. Animals treated with SALP INX 6295 exhibited a prolonged reduction of c-myc expression, reduced tumor growth and increased mice survival. When administered in combination with cisplatin (DDP), SALP INX-6295 produced a complete tumor regression in approximately 30% of treated mice, which persisted for at least 60 days following the first cycle of treatment. Finally, the median survival of mice treated with DDP/SALP INX-6295 increased by 105% compared to 84% for animals treated with the combination DDP/free INX-6295. These data indicate that the biological activity and the therapeutic efficacy of c-myc antisense therapy may be improved when these agents are administered in lipid-based delivery systems. PMID- 11498767 TI - Tissue-specific gene expression in medullary thyroid carcinoma cells employing calcitonin regulatory elements and AAV vectors. AB - Calcitonin (CT), the major secretory product of the C cell, is also expressed in C-cell-derived neoplasia. To investigate the role of the CT gene regulatory sequence in tissue-specific gene expression, the genes coding for the herpes simplex virus thymidine kinase (HSVtk) and for the enhanced green fluorescent protein (EGFP) regulated by the CT promoter (rAAV/CT266tkneo), the CT promoter/enhancer element (rAAV/CTenhtkneo), or the cytomegalovirus (CMV) promoter (rAAV/CMVtkneo) were transduced by recombinant adenoassociated viral (AAV) vectors into the medullary thyroid carcinoma (MTC) cell lines TT and hMTC and into HeLa cells as controls. In TT cell lines and hMTC cell lines transiently infected by the rAAV/CT266tkneo viruses, a significant increase in (3)H ganciclovir uptake was observed. Upon ganciclovir treatment, TT cells infected by rAAV/CT266tkneo revealed a significant growth inhibition, which was less tissue specific because HeLa cells were also affected by these particles (74.5%). In contrast, a minor but more tissue-specific growth inhibition (33.6%) was observed for TT cells after transient infection with the rAAV/CTenhtkneo particles. Employing EGFP controlled by CMV promoter and the individual CT regulatory sequences for transduction by rAAV particles, similar results were obtained indicating that both the CT promoter and enhancer element are required for tissue specific gene expression in MTC. PMID- 11498768 TI - Direct retroviral delivery of human cytochrome P450 2B6 for gene-directed enzyme prodrug therapy of cancer. AB - Human cytochrome P450 2B6 (CYP2B6) metabolizes the prodrug cyclophosphamide (CPA) to produce phosphoramide mustard that cross-links DNA leading to cell death. We have constructed a novel retroviral vector encoding CYP2B6 (designated "MetXia P450") and used it to transduce the human tumor cell lines HT29 and T47D. MetXia P450 transduction sensitised these cells to the cytotoxic effects of the prodrug CPA. Results from in vitro experiments demonstrated adverse effects on the clonogenic survival of cyclophosphamide-treated cells transduced with MetXia P450. Cytotoxic activity accompanied by bystander effect was particularly evident in 3-D multicellular spheroid models suggesting that this in vitro system may be a more appropriate model for assessing the efficacy of gene directed-enzyme prodrug therapy (GDEPT). We have applied this approach in a clinically relevant gene therapy protocol on established subcutaneous tumor xenografts. These studies show for the first time the efficacy of a P450-based GDEPT strategy mediated by a direct retroviral gene transfer in vivo. PMID- 11498769 TI - Induction of effective antitumor immune responses in a mouse bladder tumor model by using DNA of an alpha antigen from mycobacteria. AB - One of the main objectives of cancer immunotherapy is the activation and increase in number of antitumor effector cells. Recently, genetically modified tumor cell vaccines have been proposed for elicitation of antitumor effector cells. Native alpha antigen (alpha Ag) (also known as MPT59 and antigen 85B) of mycobacteria, which cross-reacts among mycobacteria species, may play an important biological role in host-pathogen interaction because it elicits various helper T-cell type 1 immune responses. To assess the induction of antitumor immune responses by alpha Ag, mouse tumor cell lines transfected with cDNA of alpha Ag from Mycobacterium kansasii were established, and the possibility of producing a tumor cell vaccine for induction of antitumor effects was explored. Transfection of tumor cell lines with an alpha Ag gene lead to primary tumor rejection and the establishment of protective immunity to nontransfected original tumor cell lines in Mycobacterium bovis bacillus Calmette-Gurin (BCG)-primed and unprimed mice. Mice immunized with tumor cell lines transfected with the alpha Ag gene showed delayed-type hypersensitivity responses in vivo and proliferative responses together with induction of interferon-gamma of spleen cells against nontransfected wild-type tumor cell lines in in vitro experiments. Moreover, immunization of mice with alpha Ag-expressing tumor cells elicited tumor-specific and cytotoxic T lymphocyte (CTL) epitope peptide-specific CD8+ CTLs. The results of this study provided evidence of the potential usefulness of alpha Ag in tumor cell vaccines. PMID- 11498770 TI - Growth of human melanoma xenografts is suppressed by systemic angiostatin gene therapy. AB - The effect of local and systemic delivery of the angiostatin gene on human melanoma growth was studied in nude mice. Liposome-coated plasmids carrying the cDNA coding for murine and human angiostatin (CMVang and BSHang) were injected weekly, locally or systemically, in mice transplanted with melanoma cells. The treatment reduced melanoma growth by 50% to 90% compared to that occurring in control animals treated with liposome-coated plasmid carrying the lacZ gene or in untreated controls. The growth of both locally injected and controlateral uninjected tumors in mice bearing two melanoma grafts was significantly suppressed after intratumoral treatment. Tumor growth inhibition was also observed in mice treated by intraperitoneal delivery, suggesting that angiostatin gene therapy acts through a systemic effect. Both melanoma growth suppression and delay in the onset of tumor growth were observed in treated mice. PCR performed on tumors and normal tissues showed that the lipofected DNA was present in tissues from treated mice, and angiostatin expression was demonstrated by RT-PCR. Histopathological analysis of melanoma nodules revealed an increase in apoptotic cells and a reduction in vessel density in tumors from treated mice. Our results suggest that systemic, liposome-mediated administration of genes coding for antiangiogenic factors represents a promising strategy for melanoma treatment in humans. PMID- 11498771 TI - Adenovirus-mediated interferon-beta gene therapy suppresses growth and metastasis of human prostate cancer in nude mice. AB - The purpose of this study was to determine the effects of interferon-beta (IFN beta) gene transfer on the growth of PC3MM2 human prostate cancer cells in nude mice. Intralesional delivery of an adenoviral vector encoding murine IFN-beta (AdIFN-beta), but not a vector encoding bacterial beta-galactosidase (AdLacZ), suppressed PC3MM2 tumors in a dose-dependent manner. At the highest dose (2x10(9) plaque-forming units, PFU), a single injection of AdIFN-beta (but not AdLacZ) suppressed orthotopic PC3MM2 tumors and development of metastasis by 80%, and eradicated the tumors in 20% of mice. Immunohistochemical staining showed that AdIFN-beta-treated tumors contained fewer microvessels, fewer proliferating cells, and more apoptotic cells than did the control tumors. Compared with controls, tumors injected with AdIFN-beta expressed higher levels of IFN-beta and inducible nitric oxide synthase (iNOS) and lower levels of basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1). In vitro analysis indicated that expression of bFGF and TGF-beta1 in PC3MM2 cells could be suppressed by the nitric oxide donor sodium nitroprusside. These data suggest that intratumoral delivery of the IFN-beta gene with adenoviral vectors could be an effective therapy for prostate cancer and that tumor suppression by AdIFN-beta correlated with up-regulation of iNOS and down-regulation of angiogenesis. PMID- 11498772 TI - Isolation, characterization, and recovery of small peptide phage display epitopes selected against viable malignant glioma cells. AB - Phage display techniques rely on nearly random oligonucleotide sequences inserted into the protein III filament binding protein of an Escherichia coli filamentous phage M13 to generate a library of phage that express more than 10(7) different peptides. Phage that expresses a sequence having high affinity for a specific molecule, cell, or tissue can then be isolated through selective binding and recovery. Selected phage cannot only be used as gene transfer vectors in themselves, but the small peptide epitopes can be sequenced and potentially recombined into the attachment proteins of viral vectors, or used by themselves to target other therapeutic agents and diagnostic imaging radiolabels. Most phage display selections are carried out against purified and/or fixed protein targets, raising concerns as to the relevance of the selected epitopes. We have selected phage from the CMTI library against viable U87-MG human malignant glioma cells using a derivation of biopanning. The library, which initially contained phage expressing 2x10(7) different epitope sequences, collapsed after four rounds of selection such that 42% of recovered clones expressed a consensus sequence. Selective binding to viable adherent U87-MG cells was subsequently demonstrated under physiologic conditions at 167% (+/-27%) unselected phage using a novel, viable enzyme-linked immunosorbent assay technique. In comparison, there was no difference in binding to control 9L rat gliosarcoma, PANC-1 human pancreatic adenocarcinoma, T98-MG human malignant glioma, or AST-4 human malignant glioma cells of selected compared to unselected phage. Using polymerase chain reaction, the epitope was recovered with flanking unique restriction sites for recombination into a herpes simplex virus type-1 vector. This study demonstrates and discusses optimized methodologies for using phage display to target viable cells. PMID- 11498773 TI - Enhanced growth suppression in esophageal carcinoma cells using adenovirus mediated fusion gene transfer (uracil phosphoribosyl transferase and herpes simplex virus thymidine kinase). AB - Advanced esophageal cancers are highly malignant and frequently resistant to 5 fluorouracil (5-FU). Escherichia coli uracil phosphoribosyltransferase (UP) is a pyrimidine salvage enzyme that alters 5-FU metabolism and sensitivity. A recombinant adenovirus encoding the UP gene (AxCA.UP) has been applied in gastric cancer gene therapy to sensitize cancer cells to lower concentrations of 5-FU. We have generated a recombinant adenovirus (AxCA.UT) encoding UP and herpes simplex virus thymidine kinase fusion protein (UT) to examine whether it would enhance the antitumor activity of AxCA.UP treatment. AxCA.UT treatment significantly enhanced the sensitivity of human esophageal cancer cells to and significantly enhanced the growth inhibition effects of UP gene therapy in vitro. Moreover, both 5-FU and ganciclovir showed bystander effects on growth inhibition. In an in vivo study, the therapeutic outcome of AxCA.UT treatment significantly enhanced the antitumor activity of AxCA.UP treatment. These observations suggest that AxCA.UT may be useful in esophageal cancer gene therapy. PMID- 11498775 TI - HSV-tk gene therapy for human renal cell carcinoma in nude mice. AB - We have treated Caki-2 human renal cell carcinoma in vivo using herpes simplex virus thymidine kinase (HSV-tk) gene therapy. Both stably transduced Caki-2 tumors, generated using retrovirus-mediated ex vivo HSV-tk gene transfer and direct intratumoral adenovirus-mediated HSV-tk gene transfer of wild type tumors, were tested. Similar treatments with LacZ containing retro- and adenoviruses were used as controls. The outcome was evaluated by imaging the tumors before and after the treatment with magnetic resonance imaging, and using histology, immunocytochemistry, and survival analysis. When implanted orthotopically into nude mouse kidneys, Caki-2 cells formed reproducible cystic papillary kidney carcinomas. In vivo magnetic resonance imaging provided an important tool for the evaluation of tumor growth. Transduction efficiency of wild-type tumors in vivo with adeno-LacZ was 22+/-14%. Significant tumor regression was achieved with direct intratumoral adeno-HSV-tk transduction followed by intraperitoneal ganciclovir (GCV) (P<.001). Also, the treatment of stably transduced Caki-2 tumors with intraperitoneal GCV resulted in a significant treatment response in the HSV-tk group as compared to the LacZ group (P<.009). Increased apoptosis and macrophage infiltrations, reduced proliferation, and degenerative changes were observed in the tumors treated with HSV-tk and GCV. Also, significant prolongation in survival was achieved with adeno-HSV-tk- and GCV-treated mice as compared to the controls. It is concluded that adeno-HSV-tk gene therapy may be useful for the treatment of renal cell carcinoma in vivo. PMID- 11498774 TI - Cathepsin L antisense oligonucleotides in a human osteosarcoma cell line: effects on the invasive phenotype. AB - Alterations in cathepsin L expression and trafficking have been associated with the progression and metastasis of several tumor entities. In the present study, we examined the effects of various cathepsin L antisense (as) phosphorothioate oligonucleotides on both the expression of cathepsin L and the invasive potential of the human osteosarcoma cell line MNNG/HOS. Seven oligonucleotides of 20-bp length each and one random control oligonucleotide were chosen to block cathepsin L expression. Northern blot analysis demonstrated a significant reduction in cathepsin L mRNA expression by the six antisense oligonucleotides at a concentration of 10 microM. Cathepsin L protein expression was reduced significantly (50-85%) by the antisense oligonucleotides, as compared with the controls. Adhesion to matrices of collagen I and matrigel was not affected. In in vitro motility and invasion assays performed in uncoated and precoated transwell chambers, the ability of cells to migrate through the filters was inhibited by 35 75% using antisense oligonucleotides. The random control did not show any inhibitory effect. These data demonstrate that in MNNG/HOS cells cathepsin L influences cellular malignancy by promoting migration and basement membrane degradation. PMID- 11498777 TI - The biology of endothelial cell-cell lateral junctions. PMID- 11498776 TI - Autocrine expression of both endostatin and green fluorescent protein provides a synergistic antitumor effect in a murine neuroblastoma model. AB - Modalities that act through different mechanisms can often provide synergistic antitumor activity for the treatment of refractory tumors when used in combination. Here we report a gene therapy approach in which the genes for the angiogenesis inhibitor, endostatin, and the marker protein and potent immunogen, green fluorescent protein (GFP), were delivered to murine neuroblastoma cells prior to inoculation of the tumor cells into syngeneic immunocompetent mice. Although the effect of either angiogenesis inhibition or immunomodulation alone resulted in only a modest delay in tumor growth, when these approaches were used in combination, prevention of the formation of appreciable tumors was effected in 15 of 24 (63%) mice. The combination of endostatin and GFP expression elicited a strong immune response that was T cell-mediated and was reactive against both GFP and tumor cell line-specific antigens. This afforded treated mice protection against subsequent tumor challenge with unmodified tumor cells. These results suggest that antiangiogenic and immunotherapy strategies, when used in a gene therapy-mediated approach, can act synergistically in an effective multimodality anticancer approach. PMID- 11498778 TI - Pores in the sieve and channels in the wall: control of paracellular permeability by junctional proteins in endothelial cells. AB - Exchange of solutes and ions between the luminal and abluminal compartments of the circulation is critically dependent on the barrier properties of the vascular endothelium. Transport of solutes and fluids occurs along the transcellular and paracellular pathways that are mediated by intracellular vesicles and intercellular junctions, respectively. Although the ability of endothelial cells to dynamically regulate permeability has long been recognized, the precise mechanism and the signaling pathways involved have not been fully elucidated. Finally, current definition of the complex molecular composition of intercellular junctions is expected to explain the difference in permeability between diverse segments of the circulation and possibly to highlight the existence of specific junctional channels. The properties of junctional adhesion molecule-1 (JAM-1) and vascular endothelial cadherin (VE-cadherin), two transmembrane components of interendothelial junctions, are described in detail. PMID- 11498779 TI - Tetraspanins and intercellular interactions. AB - The superfamily of tetraspanins comprises a group of polypeptides with four transmembrane domains that form large supramolecular structures in the plasma membrane through their associations to multiple integral membrane proteins. They are involved in homo- and heterotypic intercellular interactions in different processes such as hematopoiesis, lymphocyte activation, cancer metastasis, and fertilization. Intercellularly located tetraspanins regulate the juxtacrine activity of growth factors, cell fusion, and myelin formation. On the other hand, in motile cells they relocalize from cell-cell junctions to actin-based structures such as filopodia or growth cones and regulate cell motility in wound healing and angiogenesis processes. PMID- 11498780 TI - Roles of leukocyte and immune cell junctional proteins. AB - The restricted expression of E-cadherin on dendritic cells and intraepithelial lymphocytes has been described as a structural/adhesive system that helps to retain and integrate these cells within mucosal and dermal tissues. The activation of these cells downregulates expression of cadherins, and contributes to cell redistribution and tissue homing. It has recently been reported that lymphocytes and other leukocytes express cadherins, as well as occludin, a tight junctional component, in response to several types of stimuli. This suggests that mobilization of adherens and tight junction proteins in leukocytes may facilitate interactions of leukocytes with epithelial, endothelial, and interstitial cells that express these proteins and support homophilic adhesion. The conditions and patterns of synthesis of these adhesion molecules, in antigen-presenting cells and leukocytes, indicate that the expression of junction proteins may play roles in normal and pathological leukocyte traffic. PMID- 11498781 TI - Migration of leukocytes across endothelial junctions: some concepts and controversies. AB - This article is not meant to be a comprehensive review of leukocyte migration or endothelial cell junctions. Rather, I have chosen some aspects of inflammation that might be of general interest to vascular biologists and have focused on the structural and molecular elements of the endothelial junction involved in these processes. These are all active (and some controversial) areas of investigation. I have tried to objectively present both sides of any controversies, while stating at the end the general consensus of the field. PMID- 11498782 TI - Morphologic responses of endothelium to shear stress: reorganization of the adherens junction. AB - Shear stresses induce marked morphologic responses from endothelium which include alterations to cell shape and orientation and changes to cytoskeletal organization. These morphologic changes necessitate remodeling of cell-cell adhesion complexes that are important to control of endothelial cell physiology. Reorganization of endothelial adherens junctions has been characterized, and there are some data that pertain to the signaling pathways that regulate this reorganization. Shear-induced activation of Src, mitogen-activated protein (MAP) kinase (ERK1/2 and p38), and PI 3'-kinase pathways are important candidate pathways, and there is evidence for a role for the Rho GTPases. Very little is known concerning shear-dependence of other junctional complexes, but available data indicates a high degree of shear sensitivity. Given the continuous changes in hemodynamics which occur physiologically in vivo, sensitivity of endothelial cell-cell adhesion complexes to shear will likely prove important to vascular pathophysiology. PMID- 11498783 TI - Tissue specific expression of p53 target genes suggests a key role for KILLER/DR5 in p53-dependent apoptosis in vivo. AB - The p53 tumor suppressor plays a key role in the cell's response to genotoxic stress and loss of this 'guardian of the genome' is an important step in carcinogenesis. The ability of p53 to induce apoptosis through transactivation of its target genes is critical for its function as tumor suppressor. We have found that overexpression of p53 in human cancer cell lines resulted in apoptosis as measured by PARP cleavage. Furthermore we observed cleavage of both caspase 9 and caspase 8 after overexpression of p53 and found that p53-dependent apoptosis was inhibited by either cellular (c-Flip-s, Bcl-X(L)) or pharmacological inhibitors of caspase 8 or caspase 9 respectively. These results indicate that p53 is mediating apoptosis through both the mitochondrial and death receptor pathways. To elucidate the relevant p53 target genes and examine the caspase pathways utilized in vivo, we treated p53+/+ and age matched p53-/- mice with 5 Gy ionizing radiation or 0.5 mg/animal dexamethasone and harvested tissues at 0, 6 and 24 h. We examined the mRNA expression of p21, bax, KILLER/DR5, FAS/APO1 and EI24/PIG8 using TaqMan real time quantitative RT-PCR in the spleen, thymus and small intestine. Although the basal mRNA levels of these genes did not depend on the presence of p53, we observed a p53-dependent induction of all these targets in response to gamma-irradiation and a p53-independent regulation for p21 and KILLER/DR5 in response to dexamethasone. Furthermore, we have demonstrated that the relative induction of these p53 target genes is tissue specific. Despite observing otherwise similar levels of death in these tissues, our findings suggest that in some cases apoptosis mediated through p53 occurs by redundant pathways or by a 'group effect' while in other tissues one or few targets may play a key role in p53-dependent apoptosis. Surprisingly, KILLER/DR5 is the dominantly induced transcript in both the spleen and small intestine suggesting a potentially important role for this p53 target gene in vivo. PMID- 11498784 TI - p53 directs conformational change and translation initiation blockade of human fibroblast growth factor 2 mRNA. AB - Tumour suppressor p53 has been shown to inhibit fibroblast growth factor 2 expression post-transcriptionally in cultured cells. Here we have investigated the mechanism responsible for this post-transcriptional blockade. Deletion mutagenesis of the FGF-2 mRNA leader revealed the requirement of at least four RNA cis-acting elements to mediate the inhibitory effect of p53 in SK-Hep-1 transfected cells, suggesting the involvement of RNA secondary or tertiary structures. Recombinant wild-type, but not Ala(143) mutant p53, was able to specifically repress FGF-2 mRNA translation in rabbit reticulocyte lysate, in a dose dependent manner. Sucrose gradient experiments showed that p53 blocks translation initiation by preventing 80S ribosome formation on an mRNA bearing the FGF-2 mRNA leader sequence. Interaction of wild-type and mutant p53 with different RNAs showed no significant correlation between p53 RNA binding activity and its translational inhibiting effect. However, by checking the accessibility of the FGF-2 mRNA leader to complementary oligonucleotide probes, we showed that the binding to RNA of wild-type, but not mutant p53, induced RNA conformational changes that might be responsible for the translational blockade. This strongly suggests that p53 represses FGF-2 mRNA translation by a direct mechanism involving its nucleic acid unwinding-annealing activity. PMID- 11498785 TI - Relative frequency and morphology of cancers in carriers of germline TP53 mutations. AB - The spectrum and frequency of cancers associated with germline TP53 mutations are uncertain. To address this issue a cohort of individuals from 28 families with Li Fraumeni syndrome, segregating germline TP53 mutations was established. Predicted cancers were estimated by applying age, morphology, site and sex-specific UK cancer statistics to person-years at risk. Observed and predicted cancers were compared and two-sided P-values calculated. Cancer types occurring to excess and showing P-values <0.02, were designated strongly associated with germline TP53 mutations. These were removed from the data and a second round of analyses performed. Cancer types with P-values <0.02 and 0.02-0.05 in the second round analyses were considered moderately and weakly associated respectively. Strongly associated cancers were: breast carcinoma, soft tissue sarcomas, osteosarcoma, brain tumours, adrenocortical carcinoma, Wilms' tumour and phyllodes tumour. Carcinoma of pancreas was moderately associated. Leukaemia and neuroblastoma were weakly associated. Other common carcinomas including lung, colon, bladder, prostate, cervix and ovary did not occur to excess. Although breast carcinoma and sarcomas were numerically most frequent, the greatest increases relative to general population rates were in adrenocortical carcinoma and phyllodes tumour. We conclude that germline TP53 mutations do not simply increase general cancer risk. There are tissue-specific effects. PMID- 11498787 TI - Adenosine nucleotide modulates the physical interaction between hMSH2 and BRCA1. AB - We have identified the physical interaction between the Breast Cancer susceptibility gene product BRCA1 and the Hereditary Non-Polyposis Colorectal Cancer (HNPCC) and DNA mismatch repair (MMR) gene product hMSH2, both in vitro and in vivo. The BRCA1-hMSH2 association involved several well-defined regions of both proteins which include the adenosine nucleotide binding domain of hMSH2. Moreover, the interaction of BRCA1 with purified hMSH2-hMSH6 appears to be modulated by adenosine nucleotide much like G protein downstream interaction/signaling is modulated by guanosine nucleotide. BARD1, another BRCA1 interacting protein, was also found to interact with hMSH2. In addition, BRCA1 was found to associate with both hMSH3 and hMSH6, the heterodimeric partners of hMSH2. These observations implicate BRCA1/BARD1 as downstream effectors of the adenosine nucleotide-activated hMSH2-hMSH6 signaling complex, and suggest a global role for BRCA1 in DNA damage processing. The functional interaction between BRCA1 and hMSH2 may provide a partial explanation for the background of gynecological and colorectal cancer in both HNPCC and BRCA1 kindreds, respectively. PMID- 11498786 TI - Constitutive expression of ectopic c-Myc delays glucocorticoid-evoked apoptosis of human leukemic CEM-C7 cells. AB - Sensitivity to glucocorticoid (GC)-evoked apoptosis in lymphoid cell lines correlates closely with GC-mediated suppression of c-Myc expression. To establish a functional role for c-Myc in GC-mediated apoptosis, we have stably expressed MycER(TM), the human c-Myc protein fused to the modified ligand-binding domain of the murine estrogen receptor alpha, in GC-sensitive CEM-C7-14 cells. In CEM-C7-14 cells, MycER(TM) constitutively imparts c-Myc functions. Cells expressing MycER(TM) (C7-MycER(TM)) exhibited a marked reduction in cell death after 72 h in 100 nM dexamethasone (Dex), with 10-20-fold more viable cells when compared to the parental CEM-C7-14 clone. General GC responsiveness was not compromised, as evidenced by Dex-mediated suppression of endogenous c-Myc and cyclin D3, and induction of c-Jun and the glucocorticoid receptor. MycER(TM) also blunted Dex mediated upregulation of p27(kipI) and suppression of the Myc target p53. In comparison to parental CEM-C7-14 cells, Dex-evoked DNA strand breaks were negligible and caspase activation was delayed, but the extent of G1 cell cycle arrest was similar in C7-MycER(TM) cells. Myc-ER(TM) did not result in permanent, complete resistance to GC however, and the GC-treated cells eventually died, indicative of redundant or interactive mechanisms in the GC-evoked lytic response of lymphoid cells. Our results emphasize the importance of c-Myc suppression in GC-evoked apoptosis of CEM-C7-14 cells. PMID- 11498788 TI - Dynamic in vivo interactions among Myc network members. AB - Members of the Myc oncoprotein network (c-Myc, Max, and Mad) play important roles in proliferation, differentiation, and apoptosis. We expressed chimeric green fluorescent protein (GFP) fusions of c-Myc, Max, and three Mad proteins in fibroblasts. Individually, c-Myc and Mad proteins localized in subnuclear speckles, whereas Max assumed a homogeneous nuclear pattern. These distributions were co-dominant and dynamic, however, as each protein assumed the pattern of its heterodimeric partner when the latter was co-expressed at a higher level. Deletion mapping of two Mad members, Mad1 and Mxi1, demonstrated that the domains responsible for nuclear localization and speckling are separable. A non-speckling Mxi1 mutant was also less effective as a transcriptional repressor than wild-type Mxi1. c-Myc nuclear speckles were distinct from SC-35 domains involved in mRNA processing. However, in the presence of co-expressed Max, c-Myc, but not Mad, co localized to a subset of SC-35 loci. These results show that Myc network proteins comprise dynamic subnuclear structures and behave co-dominantly when co-expressed with their normal heterodimerization partners. In addition, c-Myc-Max heterodimers, but not Max-Mad heterodimers, localize to foci actively engaged in pre-mRNA transcription/processing. These findings suggest novel means by which Myc network members promote transcriptional activation or repression. PMID- 11498789 TI - Expression of truncated FHIT transcripts in cervical cancers and in normal human cells. AB - To analyse FHIT transcription patterns in cervical cancer, a series of primary cervical tumors and normal control samples were studied using RT-PCR. Full length and truncated FHIT transcripts were detectable in all samples tested. Interestingly, the expression of truncated FHIT transcripts by primary epithelial cells in vitro was associated with confluency. The breakpoints of most transcript deletions coincided with genuine splice site sequences, suggesting that they resulted from alternative splicing. These findings demonstrate that truncated FHIT transcripts are commonly detected in both normal and tumor tissues, and suggest that these altered transcripts are not causally related to tumorigenesis in cervical cancer. PMID- 11498790 TI - Mel-CAM-specific genetic suppressor elements inhibit melanoma growth and invasion through loss of gap junctional communication. AB - Normal human melanocytes are interspersed singly among keratinocytes along the basement membrane of the epidermis, whereas melanoma cells readily adhere to each other during invasion of the dermis or distant organs. The tumorigenic and metastatic phenotype of melanoma cells often correlates with increased expression of cell-cell and cell-matrix adhesion receptors. Mel-CAM (MCAM, MUC 18, CD146) is a cell-cell adhesion receptor highly expressed by melanoma cells but not normal melanocytes. We show here that inhibition of Mel-CAM expression in metastatic melanoma cells using genetic suppressor elements of Mel-CAM cDNA leads to inhibition of adhesion between melanoma cells and to downregulation of the tumorigenic phenotype. Growth was not inhibited in genetic suppressor elements transduced melanoma cells cultured in monolayers but was inhibited when cells were maintained anchorage-independently in soft agar and greatly reduced in immunodeficient mice. A three-dimensional epidermal skin equivalent model demonstrated that Mel-CAM allows melanoma cells to separate from the epidermis and invade the basement membrane zone and dermis. However, melanoma cells with little or no Mel-CAM were poorly invasive, possibly due to their loss of gap junctional communication. These results suggest the multifunctional role of a melanoma-associated cell-cell adhesion receptor in tumor progression. PMID- 11498791 TI - The metastasis-associated Mts1(S100A4) protein could act as an angiogenic factor. AB - The involvement of Mts1(S100A4), a small Ca(2+)-binding protein in tumor progression and metastasis had been demonstrated. However, the mechanism by which mts1(S100A4) promoted metastasis had not been identified. Here we demonstrated that Mts1(S100A4) had significant stimulatory effect on the angiogenesis. We detected high incidence of hemangiomas--benign tumors of vascular origin in aged transgenic mice ubiquitously expressing the mts1(S100A4) gene. Furthermore, the serum level of the Mts1(S100A4) protein increased with ageing. Tumors developed in Mts1-transgenic mice revealed an enhanced vascular density. We showed that an oligomeric, but not a dimeric form of the Mts1(S100A4) protein was capable of enhancing the endothelial cell motility in vitro and stimulate the corneal neovascularization in vivo. An oligomeric fraction of the protein was detected in the conditioned media as well as in human serum. The data obtained allowed us to conclude that mts1(S100A4) might induce tumor progression via stimulation of angiogenesis. PMID- 11498792 TI - The ERK-dependent signalling is stage-specifically modulated by FSH, during primary Sertoli cell maturation. AB - Primary cultures of Sertoli cells provide an interesting model to study how signalling pathways induced by a single hormone in a single cell type evolve, depending on the developmental stage. In vivo, follicle-stimulating hormone (FSH) induces proliferation of Sertoli cells in neonate and controls the subsequent differentiation of the entire population. Molecular mechanisms underlying Sertoli cell pleiotropic responses to FSH have long been investigated. But to date, only cAMP-dependent kinase (PKA) activation has been reported to account for most FSH biological activities in male. Here, we demonstrate that FSH activates the ERK MAP kinase pathway following dual coupling of the FSH-R both to Gs and to Gi heterotrimeric proteins, in a PKA- and also Src-dependent manner. This activation is required for FSH-induced proliferation of Sertoli cells isolated 5 days after birth. Consistently, we show that the ERK-mediated FSH mitogenic effect triggers upregulation of cyclin D1. In sharp contrast, at 19 days after birth, as cells proceed through their differentiation program, the ERK pathway is dramatically inhibited by FSH treatment. Taken together, these results show that FSH can exert opposite effects on the ERK signalling cascade during the maturation process of Sertoli cells. Thus, signalling modules triggered by the FSH-R evolve dynamically throughout development of FSH natural target cells. PMID- 11498793 TI - Integrin alphavbeta3 promotes anchorage-dependent apoptosis in human intestinal carcinoma cells. AB - A population of cells surviving during prolonged incubation in suspension (anoikis-negative cells) were selected from the original anoikis-positive human intestinal carcinoma cell line Caco-2. Anoikis-negative cells are characterized by a strong transcriptional downregulation of the alphav-integrin chain as detected by FACS analysis, RT-PCR and Northern blotting. This finding suggested that alphav-integrin generates a signal stimulating apoptosis of Caco-2 cells upon their detachment from the extracellular matrix. Two lines of evidence supporting this suggestion were provided. First, activation of the alphavbeta3 integrin on Caco-2 cells by their treatment with an alphavbeta3-specific monoclonal antibody resulted in marked stimulation of anoikis. Second, treatment of Caco-2 cells with alphav-specific antisense oligonucleotide resulted in downregulation of the expression of alphav chain and in elevated resistance of these cells to anoikis. Thus, for the first time, our data prove that alphavbeta3 integrin can be an active transducer of apoptosis-stimulating signals generated in response to disruption of the cell-matrix contacts. PMID- 11498794 TI - Involvement of PLAGL2 in activation of iron deficient- and hypoxia-induced gene expression in mouse cell lines. AB - We searched iron-deficient inducible cDNA, using subtraction cloning and mRNA from desferrioxamine-treated mouse macrophage Raw264.7 cells. We identified a pleomorphic adenoma gene like 2 (PLAGL2), one of PLAG superfamily proteins exhibiting antiproliferative properties on tumor cells. Mouse PLAGL2 consists of 496 amino acids with seven C2H2 zinc-fingers. PLAGL2 mRNA was induced in RAW264.7 cells, mouse erythroleukemia cells and Balb/c 3T3 cells when they were treated with desferrioxamine. Hypoxia also increased PLAGL2 mRNA. Expression of PLAGL2 in COS-7 cells led to nuclear localization. PLAGL2 had potential binding ability to GC-rich oligonucleotide and activated transcription of a gene with the binding sequence in transient reporter assay, a finding consistent with a case seen in a PLAGL2 homolog, ZAC-1. Transient co-transfection of PLAGL2 or ZAC1 cDNA and a reporter containing a lactate dehydrogenase A (LDHA) promoter carrying the hypoxia inducible factor-1 responsive element led to an increase in the basal transcription in Balb/c 3T3 and HepG2 cells. Activation in transcription from the LDHA promoter increased by desferrioxamine treatment or hypoxia was further enhanced when PLAGL2 was expressed. We propose that PLAGL2 is involved in the cell cycle arrest and apoptosis of tumor cells by regulating iron depletion- or hypoxia-inducible gene expression. PMID- 11498795 TI - Murine embryonic fibroblasts lacking TC-PTP display delayed G1 phase through defective NF-kappaB activation. AB - Previous results suggested a potential role for T-cell protein tyrosine phosphatase (TC-PTP) in cell proliferation. However, no conclusive data has supported such a function in the modulation of this process. In order to clarify this issue, we isolated TC-PTP-/- murine embryonic fibroblasts (MEFs) as well as cell lines to characterize the role of TC-PTP in the control of cell proliferation and cell cycle. Both TC-PTP-/- primary MEFs and cell lines proliferate slower than TC-PTP+/+ cells. We also demonstrated that TC-PTP-/- cells have a slow progression through the G1 phase of the cell cycle. Further characterization of the G1 defect indicates that the kinetics of cyclin D1 induction was delayed and that p27(KIP1) remains at higher levels for an extended period of time. Moreover, cells lacking TC-PTP showed a delayed activation of CDK2. This slow progression through the early G1-phase resulted in decreased phosphorylation of the RB protein and subsequent delay into the S phase transition. In contrast, no further defects were detected in other phases of the cell cycle. Survey of the potential signaling pathways leading to this delayed cyclin D1 expression indicated that NF-kappaB activation was compromised and that IKKbeta activity was also reduced following PDGF stimulation. Reintroduction of wild-type TC-PTP into the TC-PTP-/- cells rescued the defective proliferation, cyclin D1 expression, NF-kappaB activation as well as IkappaB phosphorylation. Together, these results confirm that TC-PTP plays a positive role in the progression of early G1 phase of the cell cycle through the NF-kappaB pathway. PMID- 11498797 TI - Cross-regulatory interaction between Stra13 and USF results in functional antagonism. AB - Transcription factors belonging to the basic helix-loop-helix (bHLH) family are critical regulators of cellular proliferation and differentiation. The functional activity of these proteins can be regulated by heterodimerization through the HLH domain, as a result of formation of functional or non-functional heterodimers. The presence of a leucine zipper in bHLH-leucine zipper (bHLHZip) proteins, however, prevents such heterodimeric interactions via the HLH domain between bHLH and bHLHZip proteins. To identify cellular proteins that directly interact with and modulate transcriptional repression mediated by the bHLH protein Stra13, we carried out a yeast two hybrid screen. The bHLHZip protein USF (Upstream Stimulatory factor) was identified as a Stra13 interacting protein. We demonstrate a direct interaction between Stra13 and USF that is dependent upon the C-terminal repression domain of Stra13 and the DNA-binding domain of USF. Stra13 and USF also colocalize and functionally interact in mammalian cells. Co expression of USF abrogates Stra13-mediated repression of target genes and conversely, Stra13 inhibits DNA-binding and USF-mediated transactivation. Taken together, our data demonstrate that Stra13 and USF interact physically and functionally, and identify a novel mode of cross regulatory interaction between members of the bHLH and bHLHZip families that abrogates their functional activity. PMID- 11498796 TI - Both Rb and E7 are regulated by the ubiquitin proteasome pathway in HPV containing cervical tumor cells. AB - High-risk human papillomaviruses (HPVs) are etiologically linked to human cervical and oral cancers. The E6 and E7 oncoproteins encoded by HPV target host cell tumor suppressor proteins. E6 induces proteolysis of p53 through the ubiquitin-proteasome pathway. Recent studies showed that overexpression of E7 caused proteolytic degradation of the tumor suppressor Rb. However, unlike p53, Rb is not regulated by proteolysis in normal cells. In addition, it was unclear whether in its natural context E7 regulates Rb through the ubiquitin-proteasome pathway. Therefore, we sought to determine whether Rb is regulated by the ubiquitin-proteasome pathway in HPV-containing tumor cells. We carried out a detailed analysis in Caski cells, that are derived from HPV-containing cervical cancer tissues. Studies with various protease inhibitors revealed that Rb is regulated specifically by the ubiquitin-proteasome pathway in HPV-containing cervical tumor cells. Several inhibitors of the 26S proteasome significantly increased the level of Rb in the Caski cells. Rb controls cell growth by forming complexes with the E2F-family transcription factors. Surprisingly, in spite of a significant accumulation of the hypophosphorylated form of Rb, no Rb/E2F complex was detectable in the proteasome inhibitor treated cells. Further analysis revealed that there was an increased accumulation of the E7 oncoprotein. We showed that the proteasome inhibitors simultaneously blocked the proteolysis of E7 and Rb, suggesting that E7 is also regulated by the ubiquitin-dependent proteolysis in cervical cancer cells. Taken together, this study suggests that targeted inhibition of Rb proteolysis will be required for restoring Rb function in HPV-containing cervical cancer cells. PMID- 11498798 TI - Inhalation toxicity studies of the alpha,beta-unsaturated ketones: ethyl vinyl ketone. AB - The National Toxicology Program is conducting a chemical class study to investigate the structure-activity relationships for the toxicity of alpha,beta unsaturated ketones. Ethyl vinyl ketone (EVK) was selected for study because it is a representative straight-chain aliphatic alpha,beta-unsaturated ketone with extensive use and widespread exposure. Short-term inhalation studies of EVK were conducted to provide toxicity data for comparison with the related alpha,beta unsaturated ketones 2-cyclohexene-1-one (CHX) and methyl vinyl ketone (MVK). These data will be used in designing chronic toxicity and carcinogenicity studies of these ketones. Male and female F344 rats and B6C3F1 mice were exposed to 0, 2, 4, or 8 ppm EVK 6 h/day, 5 days/wk for 13 wk. The nasal cavity was the major target organ of EVK in both rats and mice. Pathologic findings in both the olfactory and respiratory epithelium were observed. Lesions consisted primarily of olfactory epithelial necrosis, atrophy and regeneration, and/or hyperplasia and squamous metaplasia of the respiratory epithelium. Squamous metaplasia of the respiratory epithelium was present in all rats and mice exposed to 4 and 8 ppm EVK, and these lesions were more severe in rats than in mice. Few systemic effects were observed in rats and mice exposed to EVK. A transient decrease in total leukocytes due to decrements in lymphocyte and monocyte populations was present in male rats after exposure to 8 ppm for 3 and 21 days; however, this effect was not present after exposure for 13 wk. There were no chemical-related effects on micronucleus formation in mice, or on sperm motility and vaginal cytology in either species. EVK, like other alpha,beta-unsaturated ketones, is a reactive, direct-acting gaseous irritant with toxicity limited primarily to the upper respiratory tract. PMID- 11498799 TI - Inhibition of creatine kinase activity in rat brain by methyl bromide gas. AB - Rats were exposed to 290 or 495 ppm methyl bromide gas for 6 h/day, 3 times/wk for 4 to 8 wk. Creatine kinase (CK), aspartate aminotransferase (ASAT), and lactate dehydrogenase (LDH) activities and bromide ion concentrations were measured in eight regions of the brain. Methyl bromide gas inhibited CK activities in all regions of the brain, though the inhibition tended to be smallest in the cerebellum (hemisphere and vermis) and largest in the brainstem (hypothalamus, midbrain, and medulla oblongata). The dose of methyl bromide to inhibit CK activities was lower than that to damage the central nervous system histologically. No inhibition of ASAT or LDH activities was seen except for a slight inhibition of these in striatum. Inhibition of CK activities did not increase clearly on increasing dose (290 to 495 ppm) or on prolonging exposure period (4 to 8 wk). Although 50% recovery of CK activities and the half-life of bromide ion agreed well in the medulla oblongata, changes in CK activities and bromide ion concentrations did not correlate otherwise. Thus, inhibition of CK activities in brain appears to be a sensitive indicator of methyl bromide intoxication, and may be related to genesis of its neurotoxicity. The inhibition seems to be caused by methyl bromide itself rather than by bromide ion. When effects on enzyme activities in brain homogenate were examined in vitro by bubbling with methyl bromide gas, CK inhibition was seen within 15 s of exposure. Dithiothreitol suppressed the CK inhibition, whereas N-acetylcysteine did not. These observations suggest that methyl bromide may attack sites in the CK molecule different from those attacked by ethylene oxide or acrylamide. PMID- 11498800 TI - Cross-fostering inhalation toxicity study with HCFC-123 in lactating Sprague Dawley rats. AB - A study was performed in Sprague-Dawley rats (Crl:CD BR) to differentiate between effects of hydrofluorocarbon 123 (HCFC-123) on the lactating dam or on the fetus using fostering and cross-fostering of the offspring. Pregnant and/or lactating dams without the pups present were exposed to the test substance (1000 ppm) or clean air by whole-body inhalation for 6 h/day from day 6 to 19 post conceptionem (p.c.) and from day 5 to 21 post partum (p.p.). Pups were cross-fostered to new dams within the first 2 days after birth. Treatment of the mothers with HCFC-123 led to decreases in serum glucose, cholesterol, and triglycerides and increases in absolute and relative maternal liver weights. Decreased litter and individual pup weight and decreased serum triglycerides were observed in the pups of treated foster mothers. Treatment of the mothers with HCFC-123 did not influence milk production based on the body weight difference of the dam before suckling and 60 min after beginning of suckling using 12-pup "standard litters" of untreated dams. Total fat, glucose, and protein contents in the milk were also not influenced by the treatment. Trifluoroacetic acid (TFA), a main metabolite of HCFC-123, was observed in urine samples of standard litters that had been nursed by treated dams. In conclusion, the effects on offspring due to HCFC-123 treatment consisted of decreased pup weight and decreased serum triglycerides at weaning. All effects were due to treatment of the lactating dams, as no prenatally induced effects were found. Since milk production and nutritional constituents of the milk were not influenced, but significant amounts of the main metabolite were found in pup urine, an effect of HCFC-123 or its metabolite on the pups via maternal milk is considered to be a possible cause for their decreased weight gain. PMID- 11498801 TI - Recovery from oxidant-mediated lung injury: response of metallothionein, MIP-2, and MCP-1 to nitrogen dioxide, oxygen, and ozone exposures. AB - Oxidant-induced lung injury is believed to be mediated by reactive oxygen species. Recovery from oxidant exposure has been associated with pulmonary inflammation. Inflammatory cell accumulation involves the synthesis of chemokines, including neutrophil chemoattractants such as macrophage inflammatory protein-2 (MIP-2) and monocyte chemoattractants such as monocyte chemoattractant protein-1 (MCP-1). Antioxidants are the first line of defense of lung cells against inhaled oxidants. Metallothionein (MT) can act as an antioxidant and free radical scavenger. To better understand the pulmonary response associated with recovery from oxidant-mediated injury, we exposed mice to either 15 ppm nitrogen dioxide for 24 h, >99% oxygen for 72 h, or 1 ppm ozone for 24 h. Mice were examined at the end of exposure or after recovering in room air for 4 or 24 h. Neutrophils were elevated at the end of exposure and remained elevated through the postexposure period, whereas macrophage numbers were decreased at the end of exposure and remained below control levels at 4 and 24 h postexposure. MT, MIP-2, and MCP-1 mRNA levels were elevated at 4 h postexposure; however, after 24 h of recovery only MCP-1 remained elevated. These results indicate that MT, MIP-2, and MCP-1 mRNA levels responded similarly to recovery from nitrogen dioxide, oxygen, and ozone exposure. Monocyte accumulation was delayed as compared to neutrophils and was consistent with the timing of MIP-2 and MCP-1 expression. Peak expression of MT and MIP-2 preceded peak neutrophil accumulation. Consequently, the timing of MT, MIP-2, and MCP-1 expression may be important biological markers in assessing the state of injury and recovery associated with oxidant-mediated injury. PMID- 11498802 TI - Time study on development and repair of lung injury following ozone exposure in rats. AB - The aim of this study was to investigate the time course of lung injury in rats during acute and subchronic ozone exposure and during postexposure recovery. Rats were continuously exposed to 0.4 ppm ozone ( approximately 0.8 mg O(3)/m(3)) for 1, 3, 7, 28, or 56 days. Recovery from 3 days of exposure was studied at day 7, 14, and 28; recovery from 7 days of exposure was studied at day 14, 28, and 56, recovery from 28 days of exposure was studied at day 35 and 56, and recovery from 56 days of exposure was studied at day 136. The study included a correlated biochemical and morphological analysis of inflammatory responses, structural changes, and collagen content. The acute inflammatory response, as measured by an increase of polymorphonuclear cells and plasma protein in bronchoalveolar lavage (BAL) fluid, reached a maximum at day 1 and resolved largely within 6 days during ongoing exposure. Numbers of macrophages in BAL fluid increased progressively up to day 56, and slowly returned to near control levels when exposure was followed by postexposure recovery. Histological examination and morphometry of the lungs revealed centriacinar inflammatory responses throughout ozone exposure. Centriacinar thickening of septa was observed at day 7. Ductular septa, thickened progressively at days 7, 28, and 56 of exposure, showed increased collagen upon exposure at day 28, which was further enhanced at exposure at day 56. Increased collagen content in lungs, as measured biochemically by hydroxyproline concentration, was observed at exposure day 56. Collagen content was not different from control at day 56 when 7 or 28 days of exposure was followed by postexposure recovery. After continuous ozone exposure, respiratory bronchioles were present in an increasing degree, and remained present after a recovery period. The results of this study clearly show that after continuous exposure to O(3) some acute effects, such as protein and albumin content, and neutrophil influx in BAL fluid, returned to control levels within a few days. However, other parameters, such as the alveolar macrophage response and structural changes such as the presence of terminal bronchioles, thickening of ductular septa by enhanced cellularity, and collagen formation, persisted or progressively increased during continued exposure. Postexposure recovery seems to partly resolve these subchronic responses (macrophages response, septal cellularity), whereas other effects (collagen increase and respiratory bronchioles formation) do not disappear. PMID- 11498803 TI - Respiratory irritation by trimellitic anhydride in Brown Norway and Wistar rats. AB - Several acid anhydrides are known for their sensitizing and irritative properties. Since both irritation and respiratory allergy can cause changes of lung function, proper testing of allergen-dependent effects on the respiratory tract requires knowledge of the respiratory irritant effects. To study the latter effects, groups of female Brown Norway (BN) and Wistar rats were exposed for 30 min to a range of concentrations (10 to 300 mg/m(3)) of the well-known respiratory allergen trimellitic anhydride (TMA). Breathing pattern and frequency were monitored before, during, and after exposure. Animals were necropsied and lung weights were determined 1 day after exposure. In BN rats, changes in breathing pattern were seen at levels of 29 mg/m(3) and higher and decreases in frequency at 60 mg/m(3) and higher, whereas in Wistar rats changes in both pattern and frequency (increases followed by decreases) were seen at levels of 34 mg/m(3) and higher. Changes in breathing pattern consisted of a spiked form instead of a wave form of the respiratory cycle, with a pause between breaths at the end of expiration. The length of the pause increased with increasing concentrations of TMA while the duration of the respiratory cycle decreased slightly, implying that breathing frequency was mainly determined by the magnitude of the increase in pause. These reversible changes in breathing pattern and frequency were considered to be suggestive of lower airway irritation, rather than upper airway irritation. No concentration-related changes in lung weights were observed. The highest level at which no acute airway irritation as based on both breathing pattern and frequency was observed in both rat strains was 14 mg/m(3). PMID- 11498804 TI - Toxicity of fibers and particles. Report of the workshop held in Munich, Germany, 26-27 October 2000. PMID- 11498805 TI - The influence of suspension nebulization or instillation on particle uptake by guinea pig alveolar macrophages. AB - Phagocytosis represents a crucial event in the host defense against pathogens. Experimental methods are required that allow a range of particle doses to be delivered. However, it is not clear that these methods result in the same sites of deposition or mechanisms of clearance. The effect of particle administration by nebulization or instillation on the uptake by guinea pig alveolar macrophages (AMs) has been studied. Suspensions of escalating quantities of 1-microm fluorescent polystyrene latex microspheres were delivered by 15 min of nebulization (1.4 x 10(7)-11.1 x 10(7) particles) or instillation (19 x 10(7)-152 x 10(7) particles) into the lungs of guinea pigs. These doses were selected to maximize delivery using each of these methods. Macrophages were collected by alveolar lavage 6 h postadministration. The total number of cells recovered was 3 x 10(6) and the cell viability was >97%, which was measured by trypan blue exclusion. Differential cell counts of lavaged cell suspensions were conducted and results showed no difference for the two methods of administration with various concentrations of latex particles and control samples. The uptake of particles was measured using epifluorescence, confocal microscopy, and flow cytometry. AMs showed a dose-dependent increase in associated particles measured by microscopy and flow cytometry. There was a direct correlation (R(2) =.99) in the phagocytic indices (PIs) measured by flow cytometry and fluorescence microscopy. The PI was 15 times higher after instillation than that obtained after particle nebulization. The percentage of AMs involved in phagocytosis observed after instillation was 55% and after nebulization 23%. The uptake of aerosolized particles by AMs and the number of cells involved in phagocytosis were dependent on the particle dose and the efficiency of aerosol delivery to the lungs. PMID- 11498806 TI - Rat lung inflammatory responses after in vivo and in vitro exposure to various stone particles. AB - Rat lung alveolar macrophages and type 2 cells were exposed for 20 h in vitro to various stone particles with differing contents of metals and minerals (a type of mylonite, gabbro, feldspar, and quartz). The capability to induce the release of the inflammatory cytokines interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-alpha), and macrophage inflammatory protein-2 (MIP-2) was investigated. We found marked differences in potency between the various particles, with mylonite being most potent overall, followed by gabbro, and with feldspar and quartz having an approximately similar order of lower potency. The results also demonstrated differences in cytokine release pattern between the two cell types. For all particle types including quartz, type 2 cells showed the most marked increase in MIP-2 and IL-6 secretion, whereas the largest increase in TNF-alpha release was observed in macrophages. To investigate possible correlations between in vitro and in vivo inflammatory responses, rats were instilled with the same types of particles and bronchoalveolar lavage (BAL) fluid was collected after 20 h. The results demonstrated a correlation between the in vitro cytokine responses and the number of neutrophilic cells in the BAL fluid. The BAL fluid also showed a strong MIP-2 response to mylonite. However, this was the only particle type to give a significant cytokine response in the BAL fluid. We further examined whether a similar graded inflammatory response would be continued in type 2 cells and alveolar macrophages isolated from the exposed animals. Again a differential cytokine release pattern was observed between type 2 cells and macrophages, although the order of potency between particle types was altered. In conclusion, various stone particles caused differential inflammatory responses after both in vitro and in vivo exposure, with mylonite being the most potent stone particle. The results suggest the alveolar type 2 cell to be an important participant in the inflammatory response following exposure to particles. PMID- 11498807 TI - Sensory-nerve-mediated nasal vasodilatory response to inspired acetaldehyde and acetic acid vapors. AB - This study was designed to characterize the acute nasal vasodilatory responses to the sensory irritants acetaldehyde and acetic acid. For this purpose, the upper respiratory tract of the urethane-anesthetized male F344 rat was isolated by insertion of an endotracheal cannula, and irritant-laden air was drawn continuously through that site at a flow rate of 100 ml/min for 50 min. Vascular function was monitored by measuring inert vapor (acetone) uptake throughout the exposure. Both acetaldehyde and acetic acid induced an immediate concentration dependent vasodilation as indicated by increased steady-state acetone uptake rates. This response was observed at exposure concentrations of 25 ppm or 130 ppm or higher for acetaldehyde or acetic acid, respectively. The response to either vapor was significantly diminished in rats pretreated with the sensory nerve toxin capsaicin (50 mg/kg, 7 days prior to exposure), providing evidence that sensory nerves play a role in the response. Acetaldehyde is metabolized by aldehyde dehydrogenase to acetic acid. Pretreatment with the aldehyde dehydrogenase inhibitor cyanamide (10 mg/kg, 1 h prior to exposure) reduced the vasodilatory response to 200 ppm but not to 50 ppm acetaldehyde. These results suggest that formation of acetic acid is important in the sensory nerve-mediated vasodilatory response to high, but perhaps not to low, concentrations of acetaldehyde. PMID- 11498808 TI - Eastern teachings, western musings. PMID- 11498809 TI - Help-seeking for parents of individuals experiencing a first episode of schizophrenia. AB - The purpose of this triangulated study was to understand parents' experience of help-seeking in response to their child's first episode of schizophrenia. Twenty participants completed questionnaires related to stigma, parental caregiver burden, uncertainty in illness and help-seeking. Findings indicated: (1) as stigmatized attitudes towards mental illness and feelings of ambiguity increased, so did burden; (2) as parental education increased, uncertainty regarding their child's illness decreased; (3) as the number of symptoms causing distress increased so did burden and lack of clarity regarding their child's illness. A significant amount of time passed before parents were able to access help. No relationships were found between indicators of parental help-seeking and the study variables. A subset of five participants described their help-seeking experience. Three overarching themes emerged: (1) Evolving change: What does it mean? (2) Continuous help-seeking, and (3) The help-seeking experience: Impact on parents. PMID- 11498810 TI - Brain, behavior, connections and implications: psychodynamics no more. AB - The premise of this article is that the major conceptual approach to mental health in the 21st century is grounded in molecular biology. We introduce and discuss three aspects of molecular biology: the concept of neuroplasticity with attention to kindling and the sensitization effect, the effects of life events on brain and behavior, and the interaction of genes and the environment. Subsequent to each of these sections we include a discussion of the implications that these ideas have to nursing practice. In view of the implications that molecular biology has for psychiatric nursing research and practice, we conclude with a call for a re-conceptualization of the psychiatric mental health nursing enterprise and the need for a conceptual model that is comprehensive, integrative, and dynamic. PMID- 11498811 TI - To your health: implementation of a wellness program for treatment staff and persons with mental illness. AB - Persons with mental illness often have poor access to both health promotion and primary care services. Consequently, they are at greater risk for earlier mortality, and comorbid health problems that ultimately impact their community rehabilitation. The "To Your Health" program is a health promotion program, based on the states of change model that was implemented as part of a clubhouse rehabilitation program. The program used personal wellness profiles, health assessments, and personal goal setting to assess the relative wellness of clients and staff in order to increase their awareness of health-promoting behaviors. Additionally opportunities were provided to engage individuals in making changes through participation in activities that provided opportunities to try out new behaviors and to set goals that integrated changes into their lives. The initial findings are reported along with the rationale for the involvement of clients and staff, and the role of health promotion in community rehabilitation of persons with mental illness. PMID- 11498812 TI - Experiences of women healing from childhood sexual abuse. AB - Childhood sexual abuse has affected the lives of countless survivors who seek help within the health care system. Despite extensive research and clinical work, many survivors do not receive adequate help. This research aimed to add to the body of clinical knowledge by reporting on the experiences of childhood sexual abuse survivors who have achieved a measure of healing. Findings from a naturalistic study of 14 female survivors of childhood sexual abuse are presented. The character of healing, the relationship of context with healing, what facilitated healing, and what impeded healing are discussed. PMID- 11498813 TI - Differences in anxiety and role experiences between three age groups of women with heart disease. AB - Anxiety and role experiences in women may be related to age, developmental changes, social roles, and life circumstances such as cardiac disease. One hundred fifty-five women participated in a cross sectional survey on anxiety and role experiences in early middle-age, midlife, and elderly women after heart surgery. MANOVA analysis revealed significant differences for anxiety and role experiences by age. Compared with older women, early middle-age women had lower balance between role rewards and concerns and a poorer match between ideal and actual role function, which may be a contributing factor for their increased anxiety. PMID- 11498814 TI - Looking to the future. PMID- 11498815 TI - Parental competence among African American adolescent mothers and grandmothers. AB - This study describes the influence of grandmothers on parental competence among urban African American adolescent mothers, using a model adapted from Belsky's (1984) Determinants of Parenting Process Model. The sample included 53 pairs of urban African American mothers (age 16.4 +/- 1.2 years) and their mothers (age 39.2 +/- 4.3 years). All adolescents were enrolled in school, co-resided with mothers, and had infants ages 4 to 8 months. Survey instruments measured personal resources, perception of infant temperament, and contextual sources of support and stress. The outcome measure for adolescent mothers was perceived parental competence. Multiple regression analyses indicated adolescent mothers' self esteem and mastery explained 41% of the variance and grandmothers' self-esteem explained 42% of the variance in adolescent mothers' parental competence. A combined regression equation demonstrated that adolescent mothers' mastery and grandmothers' self-esteem accounted for 65% of the variance in adolescent mothers' parental competence. PMID- 11498816 TI - Respite care users who have children with chronic conditions: are they getting a break? AB - Caregiving for children with chronic illness and disabilities by family caregivers at home is widely recognized as an optimal choice for the child and family. To promote and maintain the health of the child, primary caregiver, and entire family, respite care has been established as an important caregiving resource. The purpose of this study was to describe primary caregivers' respite service use, perceptions of actual respite, and barriers to respite and respite care. A descriptive design with written survey methods and convenience sampling techniques was used. We found that, despite accessing a variety of respite services, the participants perceived limited actual respite from caregiving. Increased preparation of respite care providers and revision of available services may be means for achieving respite for primary caregivers and their families. PMID- 11498817 TI - Measuring prenatal drug exposure. AB - Prenatal drug exposure is an important pediatric health issue. However, the effects on children are not clear because of limitations in the way drug exposure is typically measured. For example, one cannot say cocaine causes a specific outcome if cocaine exposure is not measured accurately. Before we can determine the developmental outcomes associated with drug exposure, 4 measurement issues must be considered: (1) the amount of exposure varies greatly, such as from 1 to 709 g of crack per month; (2) exposure may vary by trimester; (3) exposure could be to one drug or multiple drugs; and (4) different sources of exposure data can be inconsistent (e.g., toxicology and maternal self-report). We use data from 248 families participating in an ongoing longitudinal study to provide concrete examples of these measurement issues. Both nursing researchers and practitioners must carefully attend to measurement issues when interpreting research on the effects of prenatal drug exposure. PMID- 11498818 TI - Does teaching scald burn prevention to families of young children make a difference? A pilot study. AB - This article reports a pilot project designed to ascertain what scald burn prevention practices parents of young children used and whether teaching would lead to implementation of burn-related home-safety practices. The sample (n = 49) was followed up longitudinally, and participants completed a survey during two home interviews conducted at 4-week intervals. There was a statistically significant change in the number of scald burn prevention measures (p <.001) implemented by parents after public health nurses provided teaching. Changes in parental behavior were correlated with ethnicity (p <.05). The results have implications for teaching families effective scald burn prevention strategies. PMID- 11498819 TI - Accuracy of references in four pediatric nursing journals. AB - Inaccurate references hinder retrieval of documents, may prevent researchers from examining all of the work by an author, and may result in authors not getting credit for their work. This study determines the number and types of errors in references in four widely read pediatric nursing journals. Of the 190 references examined, 79 of them contained an error, for an overall error rate of 41.6%. Major errors, which prevent the rapid retrieval of information, occurred in 28.9% of the references. Minor errors occurred at half the rate (13.7%) of major errors. Errors in the titles of articles, chapters, and books were the most common type of mistake followed by errors in authors' names. PMID- 11498820 TI - Using homeopathy for treating childhood asthma: understanding a family's choice. AB - The incidence and severity of asthma are increasing despite concerted efforts in comprehensive management. Families may be expected to look to complementary or alternative therapies (CAM) for help in treating persistent childhood asthma. One such therapy is homeopathy, a system of medicine that uses specially prepared, highly dilute substances to induce the body's self-healing in a comprehensive manner. This article describes the contrasting experiences for a family who undergoes specialty consultations with an allergist and with a homeopath. The style of the interview and the diagnostic tools used vary, as well as the basic philosophies and goals. The advantages and limitations, as well as the regulatory framework of homeopathy are explained, as evidenced by the literature. For nurses and other clinicians caring for children and families who use nonconventional therapies, the clinical implications are that these professionals need to become knowledgeable about the various alternative therapies which can complement conventional care. Families who wish to try homeopathy along with conventional care need to have open lines of communication and cooperation between their providers, both conventional and homeopathic. The care of childhood asthma may prove to benefit from clinical trials in homeopathy. PMID- 11498821 TI - Nursing workforce issues: an update, implications, and strategies for pediatric nurses. PMID- 11498822 TI - Genomics, proteomics, and nurses. PMID- 11498823 TI - The white rose. PMID- 11498824 TI - Families of Chinese children with malignancy: the factors impact on mother's anxiety. AB - This study examines the impact of the stress experienced by mothers during a child's cancer treatment. A descriptive correlation study has been designed to examine the relationships between uncertainty, sense of mastery, boundary ambiguity, and anxiety. The sample consists of 100 mothers recruited in two teaching hospitals in Taiwan. The sense of mastery was found to act as a mediator between uncertainty and anxiety, whereas uncertainty was a good predictor of boundary ambiguity. The first 2 months of treatment and the incidence of cancer recurrence represented a significant special experience for mothers. Nursing intervention to improve the mothers' sense of mastery and to assist families in establishing functional patterns of parent-child interaction is discussed. PMID- 11498825 TI - Role of adjuvant endocrine therapy in early-stage breast cancer. AB - The value of adjuvant endocrine therapy in saving lives of women with estrogen receptor-positive (ER(+)) early-stage breast cancer cannot be disputed. Tamoxifen has proven to be effective in improving relapse-free and overall survival in both pre- and postmenopausal women with ER(+) early-stage breast cancer. In the meta analysis of the Early Breast Cancer Trialists' Collaborative Group, the proportional reduction in recurrence and mortality for 5 years of tamoxifen therapy was 50% and 28% respectively for patients with ER(+) tumors. These reductions in recurrence and mortality were similar in both lymph node-negative (N(-)) and lymph node-positive (N(+)) patients and translate to an absolute improvement in 10-year survival of approximately 11% in N(+) patients and 6% in N(-) patients. Current data suggest that about 5 years of tamoxifen therapy is the optimal duration of treatment. For women with ER(-)/progesterone receptor negative (PR(-)) tumors, tamoxifen does not lower the risk of distant metastases or improve survival. In ER(+) patients, the addition of tamoxifen to chemotherapy further lowers the risk of recurrence by about 30% to 40% when compared to chemotherapy alone. In premenopausal women with ER(+) breast cancer, ovarian ablation has proven to be as effective as chemotherapy in improving both relapse free and overall survival and the potential additive role of ovarian ablation to chemotherapy and/or tamoxifen is presently being explored in clinical trials. The combination of tamoxifen and ovarian ablation is currently being tested and may be superior to tamoxifen alone. In addition, newer, more effective, and less toxic aromatase inhibitors are also being evaluated in clinical trials in the adjuvant setting and have great promise. "Pure" antiestrogens or selective estrogen receptor down-regulators (SERDs) will be studied in adjuvant clinical trials in the near future. Recent data also suggest that molecular markers such as HER-2/neu may predict the response to endocrine therapy, and other predictive factors are currently being evaluated. Lastly, there is renewed interest in neoadjuvant endocrine therapy, a treatment option that may select those patients with early-stage breast cancer most likely to benefit from endocrine therapy. PMID- 11498826 TI - Ovarian ablation as adjuvant therapy for breast cancer. AB - For more than 100 years ovarian ablation has been used as treatment for breast cancer. There are several methods of ovarian ablation including surgical oophorectomy, radiation-induced ablation, and chronic use of luteinizing hormone releasing hormone (LHRH) analogs. In addition, there is some suggestion that cytotoxic chemotherapy may act in part by inducing ovarian ablation in premenopausal breast cancer patients. Of the numerous case series and clinical trials of ovarian ablation performed in the past century, many have been fraught with methodologic problems. The Early Breast Cancer Trialists' Collaborative Group (EBCTCG) meta-analysis of 12 properly randomized trials shows a significant improvement in disease-free survival (DFS) and overall survival for women who underwent ovarian ablation as adjuvant therapy compared with those who did not. However, a number of questions remain. The relative efficacy of chemotherapy and ovarian ablation and the value of combining ovarian ablation with chemotherapy or other endocrine therapy have not yet been determined. This articles reviews and compares the methods of ovarian ablation, and discusses the EBCTCG overview data, as well as the newer and ongoing trials, which may answer the remaining questions about the optimal use of this therapy in the adjuvant treatment of breast cancer. PMID- 11498827 TI - HER-2 and choice of adjuvant chemotherapy in breast cancer. AB - The amplification or overexpression of HER-2 is a recognized prognostic marker that is associated with poor survival for patients with node-positive breast cancer. Several studies have demonstrated that HER-2 may serve to direct the selection of optimal adjuvant chemotherapy. This article provides a critical review of the studies that offer evidence for the role of HER-2 as a predictor of response to chemotherapy. PMID- 11498828 TI - Cancer gene and oncolytic virus therapy. AB - By far, cancer accounts for the majority of gene therapy trials that are being carried out worldwide. Seventy percent of the gene therapy protocols that have been reviewed by the National Institutes of Health Recombinant DNA Advisory Committee (NIH RAC) are for the treatment of cancer. Of these, two thirds involve immunotherapy, with transfer of genes for cytokines, immune accessory molecules, or tumor antigens into a variety of cellular targets. Other clinical protocols include chemoprotection, prodrug activation, or tumor-suppressor gene replacement. Either local or distal bystander effects may mediate antitumor effects. These bystander mechanisms may help to overcome poor transduction efficiencies by currently available vectors. Replicating oncolytic viruses entering the clinic include herpes virus, Newcastle disease virus, reovirus, and others. These viruses have been shown to replicate selectively in cancer cells, albeit by different mechanisms. Reovirus, for example, requires the presence of an activated Ras signaling pathway in order to replicate and destroy cells. Tumor selectivity can be achieved by placing an essential viral gene under the control of a tumor-specific promoter. The tumoricidal effects of replicating viruses may be enhanced by genetic modification-for example, by the insertion of a cytokine gene to elicit antitumor immunity. Clearly, much work needs to be done both in the laboratory and in the clinic in order to exploit the full potential of these novel gene and viral therapies. PMID- 11498830 TI - Adjuvant chemotherapy for breast cancer: an update. AB - Adjuvant chemotherapy in breast cancer has clearly been shown to reduce mortality. The benefits extend to pre- and postmenopausal women and those with node-negative, node-positive, estrogen receptor (ER)-positive, and ER-negative disease. Updated data regarding chemoendocrine therapy in postmenopausal women and anthracycline-based regimens are presented. Dose intensity, dose density, and high-dose therapy have not been proven efficacious to date, but further trials are pending. The incorporation of taxanes and bisphosphonates has been further elucidated, with follow-up studies in progress. The potential role of trastuzumab is the focus of several clinical trials. Recent findings regarding the long-term side effects of adjuvant therapy are reviewed. These risks may outweigh the benefit derived from chemotherapy in a subset of patients who have excellent overall survival (OS) with small tumors, good prognostic features, or favorable histologic subtypes. This report will review the current state of adjuvant chemotherapy for early-stage breast cancer. PMID- 11498829 TI - New cytotoxic agents and schedules for advanced breast cancer. AB - Cytotoxic chemotherapy is important for treatment of women with hormone insensitive or hormone-refractory advanced breast cancer. A variety of agents are effective, alone or in combination. The clinical activity and side effects of many agents, as well as principles for use of chemotherapy, are reviewed. Recent advances in chemotherapy for breast cancer include important studies on the role of dose-intensity, modifications of available agents to reduce side effects, and the availability of oral chemotherapeutics. Finally, the combination of chemotherapy with novel biological agents may improve outcomes for women with certain types of breast cancer. The growing availability of such biological therapies given in combination with chemotherapy may mean better survival in the future for women with advanced breast cancer. PMID- 11498831 TI - High-dose chemotherapy and hematopoietic stem cell transplantation for breast cancer: past or future? AB - Given that each year in the United States 180,000 new cases of breast cancer are diagnosed, with about 44,000 women succumbing to the disease, and that breast cancer is the second leading cause of cancer-related death in women, it is clear that existing therapy fails a large number of patients. Recently, a number of novel strategies have been developed in attempts to improve survival. These include agents used at very high dose requiring stem cell support. High-dose chemotherapy (HDC) with hematopoietic stem cell transplantation (HSCT), most frequently in the form of peripheral blood progenitor cell transplantation (PBPCT), is an highly active treatment approach in appropriate patients and the current data relating to this modality will be reviewed here. This article will attempt to place the recent randomized studies in perspective, to highlight the strengths and limitations of the data, and to offer some thoughts on future directions for the field. PMID- 11498832 TI - Preoperative (neoadjuvant) chemotherapy in patients with breast cancer. AB - The concept of using preoperative chemotherapy in patients with operable breast cancer originated from experimental and clinical observations, as well as from theoretical hypotheses on tumor cell growth and dissemination. Results from nonrandomized studies with different chemotherapeutic agents or combination regimens given preoperatively demonstrated substantial clinical response rates but low pathologic tumor response rates. In addition, several such studies were able to show that-by reducing primary breast tumor size-preoperative chemotherapy can lead to an increase in the rate of breast-preserving procedures. Although nonrandomized studies provided useful clinical information about the effect of preoperative chemotherapy on primary breast tumors and on axillary nodes involved with tumor, they could not address the relative efficacy of preoperative versus postoperative (adjuvant) chemotherapy on disease-free survival (DFS) and overall survival (OS). As a result, several randomized trials were implemented to address the above questions. Some of the earlier trials, however, were not designed as straightforward comparisons of preoperative versus postoperative chemotherapy and, thus, did not provide meaningful answers to the fundamental question of whether DFS and OS can be prolonged by the administration of chemotherapy before surgery rather than after. The National Surgical Adjuvant Breast and Bowel Project (NSABP) B-18 trial was the largest randomized trial that aimed to compare preoperative to postoperative chemotherapy in operable breast cancer. Results from this trial on the effect of preoperative chemotherapy on local-regional disease and outcome are presented. The potential advantages and disadvantages of each approach, as well as surgical considerations and current and future directions in the use of preoperative chemotherapy, are also discussed. PMID- 11498833 TI - Prevention of invasive breast cancer in women with ductal carcinoma in situ: an update of the National Surgical Adjuvant Breast and Bowel Project experience. AB - The National Surgical Adjuvant Breast and Bowel Project (NSABP) conducted two sequential randomized clinical trials to aid in resolving uncertainty about the treatment of women with small, localized, mammographically detected ductal carcinoma in situ (DCIS). After removal of the tumor and normal breast tissue so that specimen margins were histologically tumor-free (lumpectomy), 818 patients in the B-17 trial were randomly assigned to receive either radiation therapy to the ipsilateral breast or no radiation therapy. B-24, the second study, which involved 1,804 women, tested the hypothesis that, in DCIS patients with or without positive tumor specimen margins, lumpectomy, radiation, and tamoxifen (TAM) would be more effective than lumpectomy, radiation, and placebo in preventing invasive and noninvasive ipsilateral breast tumor recurrences (IBTRs), contralateral breast tumors (CBTs), and tumors at metastatic sites. The findings in this report continue to demonstrate through 12 years of follow-up that radiation after lumpectomy reduces the incidence rate of all IBTRs by 58%. They also demonstrate that the administration of TAM after lumpectomy and radiation therapy results in a significant decrease in the rate of all breast cancer events, particularly in invasive cancer. The findings from the B-17 and B-24 studies are related to those from the NSABP prevention (P-1) trial, which demonstrated a 50% reduction in the risk of invasive cancer in women with a history of atypical ductal hyperplasia (ADH) or lobular carcinoma in situ (LCIS) and a reduction in the incidence of both DCIS and LCIS in women without a history of those tumors. The B-17 findings demonstrated that patients treated with lumpectomy alone were at greater risk for invasive cancer than were women in P-1 who had a history of ADH or LCIS and who received no radiation therapy or TAM. Although women who received radiation benefited from that therapy, they remained at higher risk for invasive cancer than women in P-1 who had a history of LCIS and who received placebo or TAM. Thus, if it is accepted from the P-1 findings that women at increased risk for invasive cancer are candidates for an intervention such as TAM, then it would seem that women with a history of DCIS should also be considered for such therapy in addition to radiation therapy. That statement does not imply that, as a result of the findings presented here, all DCIS patients should receive radiation and TAM. It does suggest, however, that, in the treatment of DCIS, the appropriate use of current and better therapeutic agents that become available could diminish the significance of breast cancer as a public health problem. PMID- 11498834 TI - Genetic susceptibility for breast cancer--risk assessment and counseling. AB - The incorporation of genetic information into clinical oncology practice is a process in evolution, necessitating consideration of cancer risk assessment, ethical and social implications of testing and efficacy of treatment, and prevention interventions. Although most cancers do have a hereditary component, recognizing individuals who are at unusually high risk is important for planning their surveillance practices and considering options for risk management over a lifetime, and because of implications for their family members. Genetic information has enormous potential to inform and transform cancer risk identification, risk reduction, and treatment practices. In this review, we summarize basic information about breast cancer genetics, examine accumulating data about the prevalence and penetrance of deleterious mutations and management options for BRCA1 and BRCA2 mutation carriers, and discuss some of the counseling issues that may occur as physicians and patients explore this exploding area of oncology. The rapid incorporation of careful genetic testing into oncology practice is an important step toward more precise risk management. Currently, the time necessary for careful discussion of the complex issues raised by genetic testing for inherited breast/ovarian cancer susceptibility may necessitate referral to geneticists or genetic counselors for truly informed consent to be obtained. However, identification of women for whom testing is appropriate and management of cancer risk with women after testing, are critical new functions for oncologists. They are only the beginning of exciting new opportunities in cancer risk and prevention. PMID- 11498835 TI - [In vitro study on the effects of the novel retinoids combined with IFN-gamma on the proliferation and differentiation of fresh acute monocytic leukemic cells]. AB - OBJECTIVE: To investigate the prospect for clinical use of novel retinoids YS 904012 and R 9158 in combination with IFN-gamma in inducing differentiation of acute monocytic leukemia. METHODS: The effects of novel retinoids with IFN-gamma on the differentiation and clonal proliferation of fresh leukemia cell from 6 monocytic leukemia patients were studied in vitro. Cell morphological examination, nitro tetrazolium blue reduction test and DCE, expression of CD14 and CD68 and CFU-L were used in the study. RESULTS: The primary leukemia cell, cultured with the combination of YS 904012 and IFN-gamma, became more mature in morphology. The NBT reduction rate and DCE were increased from 23.4% and 25.0% to 61.5% and 52.0%, respectively. The expression of CD14 and CD68 was increased. The growth of the leukemic colony in semi-solid culture was markedly inhibited. The activity of YS 904012 combined with IFN-gamma in inhibiting proliferation and inducing differentiation of monoblast was greater than that of R9158 or all-trans retinoic acid in combination with IFN-gamma (P < 0.01). CONCLUSION: The YS 904012 combined with IFN-gamma is worthy of further study for clinical differentiation therapy of acute monocytic leukemia. PMID- 11498836 TI - [Immunophenotyping in myelodysplastic syndromes]. AB - OBJECTIVE: To evaluate the values of immunophenotyping in the diagnosis and prognosis of myelodysplastic syndromes (MDS). METHODS: Surface markers of mononuclear marrow cells were analyzed by indirect immunofluorescence for a panel of monoclonal antibodies in 32 patients with MDS. RESULTS: Expression of myeloid antigen increased significantly while lymphoid antigen decreased. The antigen expression was correlated with the FAB subtype of MDS, Immature myeloid cells (CD13+, CD33+) was being increased and the lymphoid antigen expression decreased with the RA/RAS evolving to RAEB and RAEB-t. Compared with normal marrow, the number of CD34+ cells was not significantly different in RA/RAS subtypes but significantly higher in RAEB/RAEB-t subtypes. The expression of early marrow cell antigens (CD38, HLA-DR, CD99R and CD9) was significantly increased. The expression of CD34 and other early antigen was predictive for transformation and poor survival. CONCLUSION: Surface marker analysis of mononuclear marrow cells may be a useful tool for the diagnosis and prognosis of MDS. PMID- 11498837 TI - [Expression of transferrin receptor and ferrochelatase mRNAs in MEL cells]. AB - OBJECTIVE: To investigate the changes of transferrin receptor (TfR) mRNA and ferrochelatase mRNA levels under different iron status of DMSO induced or non induced MEL cells. METHODS: MEL cells were induced with 1.5% DMSO. TfR mRNA and ferrochelatase mRNA were assayed by Northern blot after adding transferrin, deferrioxamine, anti-TfR monoclonal antibody (McAb) or Epo, respectively. RESULTS AND CONCLUSION: 1. TfR mRNA level increased with cell proliferation in non induced MEL cells, while in the induced MEL cells, although the proliferating capacity was losing with cell maturation, TfR mRNA level increased with hemoglobin synthesis and its amplitude was higher than that in non-induced cells. 2. In both non-induced and induced MEL cells, TfR mRNA levels increased when intracellular iron level decreased; and vice versa. 3. Stimulation of erythropoiesis by Epo increased the expression of TfR mRNA, indicating that its expression in erythroid cells was regulated by the level of intracellular iron and the synthesis of heme. 4. The expression of ferrochelatase mRNA had no change in non-induced MEL cells, but increased significantly in induced MEL cells. 5. The level of intracellular iron in non-induced and induced MEL cells had less effects on the expression of ferrochelatase mRNA. PMID- 11498838 TI - [The clinical and biological significance of megakaryocytic antigen expression in acute myeloid leukemia]. AB - OBJECTIVE: To explore megakaryocytic (MK) antigen expression in previously untreated adult acute myeloid leukemia (AML) and its relation to the clinical and biological characteristics. METHODS: Two hundred and eleven cases of AML were detected with flow cytometric immunofluorescence assay. RESULTS: Twenty-seven cases (12.8%) were MK antigen positive with the higher positive rates in hybrid acute leukemia (45.5%) and acute monoblastic leukemia (24.1%). MK antigen expression was significantly correlated with CD34 antigen expression, high white cell count, high P-glycoprotein positivity and had no correlation with chromosome aberration. 33.3% of MK positive AML patients achieved complete remission which was significantly lower than that (71.9%) in MK negative cases. CONCLUSION: MK antigen positive AML might derived from malignant transformation of hemotopoietic stem cell at earlier stage and the detection of MK expression was of values in predicting treatment effect and prognosis for adult AML. PMID- 11498839 TI - [Study on the expression of interleukin-8 and its receptors in acute leukemia]. AB - OBJECTIVE: To evaluate the expression of interleukin 8 (IL-8) and its A, B type receptors (IL-8RA, IL-8RB) in acute leukemia (AL). METHODS: Plasma IL-8 concentrations in peripheral blood and IL-8R expressions on bone marrow mononuclear cell (MNC) membrane of 77 newly diagnosed AL patients were assayed by ELISA and FACS, respectively. IL-8 concentration in cerebral spinal fluid (CSF) of 15 AL patients in complete remission (CR) were kinetically measured. RESULTS: Plasma IL-8 levels in newly diagnosed AL patients were increased. IL-8 levels were higher in AML than in ALL, in AML-M4, M5 were higher than in AML-M1-M3, and in B-ALL were higher than in T-ALL, respectively (P < 0.05). In ALL, CR rate in patients with IL-8 > 100 ng/L was lower than in those with IL-8 < or = 100 ng/L (P < 0.05). 36.36% of the patients were MNC IL-8R positive. Peripheral blood WBC and blasts amounts in IL-8R(+) group were significantly higher than IL-8R (-) group (P < 0.05). CSF IL-8 levels in CR patients were not different from newly diagnosed patients (P > 0.05) and were increased while central nervous system leukemia (CNSL) developed. CONCLUSION: Detection of IL-8 and IL-8R might help to identify AL types and predict prognosis and the development of CNSL. PMID- 11498840 TI - [Expression of bcl-2 gene in the evolution of chronic myelogenous leukemia to blast crisis and its implication]. AB - OBJECTIVE: To investigate the expression of bcl-2 gene and cell apoptosis and cell cycle in bone marrow of chronic myelogenous leukemia (CML). METHODS: APAAP assay and in situ hybridization were used for the expression of BCL-2 protein and bcl-2 mRNA in fresh bone marrow samples from 60 cases of CML. Flow cytometry was used to assess the extent of apoptosis and cell cycle percentage. RESULTS: The expression of bcl-2 gene had no statistical difference between CMLs at presentation and in chronic phase, but was much lower in blast crisis (P < 0.05). The percentage of bcl-2 mRNA positive cells was consistent with BCL-2 protein expression. In addition, BCL-2 protein was related to the Hb levels, BPC, and immature cells in the peripheral blood and bone marrow. Notably, the extent of apoptosis in accelerated phase/blast crisis was much lower than that in chronic phase(P = 0.028), while the cell cycle had no difference. CONCLUSION: High level of bcl-2 gene expression and low extent of apoptosis in bone marrow cells of CML might partially be the mechanism of poor prognosis of blast crisis, and this provides a new experimental basis for early diagnosis and treatment of CML blast crisis. PMID- 11498841 TI - [Study on the sensitization of acute myeloid leukemia cell to daunorubicin by recombinant human interleukin-3]. AB - OBJECTIVE: To evaluate the effect of recombinant human interleukin-3(rhIL-3) on the sensitization of acute myeloid leukemia(AML) cells to daunorubicin(DNR). METHODS: AML cells were cultured in a DNR-containing medium supplemented or not with rhIL-3. CFU-AML was assayed by semi-solid culture, proliferation and differentiation of AML cells were assayed by flow cytometry, cytomorphology and benzidine staining. Intracellular DNR concentration was measured by spectrophotofluorometry. RESULTS: rhIL-3 could enhance the proliferation but not differentiation of AML cells in vitro, and had no effect on DNR intake and exclusion of AML cells while enhancing their sensitivity to DNR. CONCLUSION: rhIL 3 combined with DNR might improve the therapeutic effect of AML. PMID- 11498842 TI - [The hemoprotective effect of platelet factor 4 (PF4) and tetrapeptide AcSDKP]. AB - OBJECTIVE: To study the effects of platelet factor 4(PF4) and tetrapeptide N acetyl-Ser-Asp-Lys-Pro(AcSDKP) on hemopoietic progenitors in mice treated with 5 Fluorouracil (5-FU). METHODS: Mice were injected with PF4 (40 micrograms/kg) or AcSDKP (4 micrograms/kg) twice at 6 h intervals, and 20 h after the second injection they were given one injection of 5-FU (150 mg/kg), and the high proliferative potential-colony forming cell (HPP-CFC), burst-forming unit erythroid (BFU-E), colony forming unit megakaryocyte (CFU-MK), and megakaryocyte (MK) were examined 6, 8 and 13 days later. RESULTS: The administration of PF4 or AcSDKP resulted in a significant increase of the number of HPP-CFC on days 6-8 and BFU-E and CFU-GM on day 8 when compared to 5-FU alone. Furthermore, PF4 was found to increase significantly the number of CFU-MK and MK on day 8, which was not observed with AcSDKP. CONCLUSION: PF4 or AcSDKP accelerate the recovery in vivo of HPP-CFC, CFU-GM and BFU-E after 5-FU treatment but their effect may be different on megakaryocytic progenitors. Both molecules may have a hemoprotective effect against chemotherapeutic agents. PMID- 11498843 TI - [Analysis of fibronectin and laminin in the extracellular matrixes of acute myeloid leukemia and its implications]. AB - OBJECTIVE: To investigate the relationship between quantitative change of fibronectin (Fn) or Laminin (Lm) and growth and differentiation of leukemic cells. METHODS: The extracellular matrixes, Fn and Lm, in long-term bone marrow culture (LTBMC) from 18 acute myeloid leukemia (AML) patients and 6 normal subjects were examined by immunofluorescence assay. RESULTS: 1. The Fn or Lm content in 9 AML self-maintained (AMLsm) 1-3 week-cultures was significantly less than that in 9 AML not self-maintained (AMLnsm) cultures. 2. In the third week's cultures, the blasts of 8 AMLnsms decreased to less than 5%, but did not of 7 AMLsms. 3. As the cultures going on, the blasts were related to Fn or Lm contents positively in normal control and negatively in AMLnsm. The blasts were related to Fn contents negatively in AMLsm. CONCLUSION: The change of Fn or Lm may play an important role in the development of AML. In addition, the difference of Fn or Lm contents between AMLsm and AMLnsm was helpful to the choice of AML patient for in vitro purging of leukemic cells through LTBMC before auto-BMT. PMID- 11498845 TI - [Enhance the study of diagnosis and treatment of acute leukemia at molecular level]. PMID- 11498844 TI - [Progress on the study of related genes in patients with blast crisis of chronic myelocytic leukemia]. PMID- 11498846 TI - [Relationship between the bone marrow cell proliferation and the prognosis in childhood acute lymphoblastic leukemia]. AB - OBJECTIVE: To study the expression of proliferative antigens in leukemic cells and the relationship between the cell proliferation activity and the prognosis. METHODS: The labeling index (LI) of CD71, Ki-67 and PCNA in normal and leukemic cells were measured by flow cytometry. RESULTS: In normal children, the LI of CD71, Ki-67 and PCNA was (32.18 +/- 16.66)%, (4.82 +/- 9.27)% and (19.69 +/- 25.11)%, respectively, while in ALL children, which was (33.66 +/- 21.52)%, (32.14 +/- 23.59)% and (47.46 +/- 30.96)%, respectively. The PCNA LI of leukemia cells was (47.46 +/- 30.96)% and (27.28 +/- 12.51)% when patients was at presentation and remitted for 3 years, respectively, and was (52.59 +/- 32.00)% and at presentation in 28 CCR children and and (26.94 +/- 14.48)% in 7 high-risk group ALL children (F = 8.877, P < 0.01), respectively. CONCLUSIONS: The proliferation, LI of leukemic cells was higher than that of normal cells, and reduced to normal level after treatment. The higher PCNA proliferation of untreated cells was a favourable marker for prognosis. PMID- 11498847 TI - [Psammomatoid ossifying fibroma]. AB - OBJECTIVE: To clarify the histopathological, clinical behavior biological characterics and the relationship among the psammomatoid ossifying fibroma (POF), ossifying fibroma and fibrous dysplasia of bone. METHODS: Thirty-one cases of POF in the ear and sinonasal region were studied retrospectively. The sections, clinicopathological records from June, 1956 to November, 1996 were reviewed. The reticulum fiber and Masson's stain and alkaline phosphatase immunostain were done in 10 cases, Vimentin, Desmin, EMA and smooth actin immunohistochemical stain were done in 5 cases. One case was studied by transmission electron microscope. RESULTS: The specimens were broken tissue with the sand-like felling, eleven specimens with intact thin shell consisted of bone tissue were used to study the changes in the area between tumor and normal bone. Histopathologically they were rich in mesenchymal tumor cells and a lot of calcifying psammomatoid-like osteoid and a few of calcified bodies similar to those described by Shafer, and the bony trabecula was not found in the center of the tumor. In addition, three zones were noted at the junction of normal bone and tumor, namely, the normal bone-reactive proliferative bone-tumor. In some cases, the reactive bony proliferation may be not marked. The normal bone adjacent to the tumor may be absorbed and disappeared, and were then replaced by the tumor, and finally led to tumor tissue invasion into the soft tissue, but the infiltration appearance is not present. CONCLUSION: POF is the slowly growing benign tumor and is different from ossifying fibroma and fibrous dysplasia of bone. PMID- 11498848 TI - [Salvage surgery for nasopharyngeal carcinoma after irradiation failure]. AB - OBJECTIVE: To study the salvage surgery for nasopharyngeal carcinoma after irradiation failure. METHODS: One hundred and twenty-one full-dose-irradiation uncontrolled cases of nasopharyngeal carcinoma had undergone salvage surgery since 1965 till 1992, in which 41 had primary lesion and 80 neck lesion. All patients received radiation before surgery with the dose of 60Gy-145Gy. The patient selection for this report limited to those with recurrent tumors at the primary sites with no apparent bony involvement and no cranial nerve palsy, or cases with resectable neck recurrences. RESULTS: It showed that palatal fenestration was the best choice of approach for recurrent tumors located at the vault of the nasopharyngeal cavity. When the tumor invaded the parapharyngeal space or, owing to the after-effect of radiotherapy, the patient presented with severe occlusion of the mandible, we preferred the lateral approach through partial mandi-bulectomy. The lateral rhinotomy was only indicated for cases with tumor remnant at the posterior nares. Lymph node excision was performed for cases with single movable node on the neck, and RND for the cases with multiple nodes. The complication rate of after surgery was low (10%), considering the heavy dose of irradiation. The three- and five-year survival rate were 49% (59/121) and 38% (38/100) respectively. CONCLUSION: It suggests that the surgery should be considered as a salvage procedure for cases of nasopharyngeal cancer after irradiation failure. Because of high rate of distant metastasis (62 cases or 45% died after surgery), chemotherapy or other measures should be adopted after local treatment. PMID- 11498849 TI - [Preliminary study on the correlation between neovasculature and metastatic nasopharyngeal carcinoma by computer image analysis]. AB - OBJECTIVE: To investigate the significance of neoangiogenesis in metastasis of nasopharyngeal carcinoma (NPC) so as to clarify further the role of neovascularity in evaluating the prognosis of NPC and the probability of antiangiogenesis in preventing distant metastasis of NPC. METHODS: Fifty-two patients with metastatic NPC were selected and strictly matched in sex, age, T stage, and N stage with another 52 patients with non-metastatic NPC and survived a long time. The tumor tissues of all 104 patients were retrived for computer assisted, immunohistochemical analysis of tumor vasculature. RESULTS: Counts of microvessel and relative area of all microvessels per image were significantly higher in metastatic NPCs than those of curable, non-metastatic NPCs; while average area of microvessel and average perimeter of microvessel in metastatic NPCs were smaller than those of the opposite. No significant difference in any microvessel parameter had been found among various types of metastasis. CONCLUSIONS: The alterations of microvessel parameters bore significant relationships with metastases of NPC. Evaluation of neovascularity with computer image analysis may help in estimating the prognosis of NPC and in finding out the indications for aggressive multimodal treatments. Angiogenesis inhibitors might be helpful in preventing NPC from distant metastasis. PMID- 11498850 TI - [Loss of heterozygosity on chromosome RB1.20 locus and p110RB1 protein state in squmous cell carcinomas of larynx]. AB - OBJECTIVE: To study the RB1 gene in squamous cell carcinoma of larynx(ISCC) and to find clue for discovering and locating new suppressor gene. METHODS: Loss of heterozygosity (LOH) of microsatellite polymorphic sequence on chromosomes 13 at RB1.20 locus of 58 LSCC patients were analyzed. The p110RB1 protein state was detected by immunohistochemical staining using the polyclonal antibody to the RB1 products. RESULTS: It showed that 3 cases in the preinvasive stage (i.e. carcinoma in situ) had no any LOH on chromosome 13. Forty percent of the 55 invasive LSCC showed LOH at RB1.20 locus. By immunohistochemical staining p110RB1 protein negative reaction was observed in 8 LSCCs in which 6 cases associated with LOH at RB1.20 locus. CONCLUSIONS: RB1 gene was inactivated in some LSCCs. It is putative that there is another tumor suppressor gene on chromosome 13 near by RB1.20 locas. The inactivation of the genes at the chromosome 13q region including RB1 involved genesis and development of invasive LSCC. PMID- 11498851 TI - [Nonepithelial malignant neoplasms of the larynx and hypopharynx]. AB - OBJECTIVES: Nonepithelial Malignant Neoplasms of the larynx and hypopharynx are rare. Their clinical features, diagnosis and treatment were discussed. METHODS: From 1985 to 1996, 23 cases of nonepithelial malignant neoplasms of the larynx and 3 cases of hypopharynx were diagnosed. There were 21 males and 5 females. Age of the patients ranged from 16 to 65 years. Laryngeal tumours included 11 cases of sarcomas, 1 extramedullary plasmocytoma, 9 lymphomas and 2 malignant melanomas. Hypopharyngeal tumours included 1 leiomyosarcoma and 2 malignant melanomas. Extensive resection was the first treatment for 21 patients. Then, according to their histopathologic features, irradiation or/and chemotherapy were chosen. RESULTS: Follow-up period was from 6 months to 10 years. There were 12 patients survived for 4 to 10 years, including 6 cases of laryngeal sarcomas, 2 supraglottic lymphomas, 2 malignant melanomas of the larynx, 1 malignant melanoma and 1 leiomyosarcoma of the hypopharynx. CONCLUSIONS: Characteristics for these tumours were: 1. usually covered by intact mucosa; 2. laryngeal tumours rarely metastasize to the cervical lymph nodes; 3. difficulties in histopathological diagnosis, and immunohistochemistry is helpful to differentiation and determination. PMID- 11498852 TI - [Preoperative nasal continuous positive airway pressure treatment as substitute for protective tracheostomy in severe obstructive sleep apnea syndrome]. AB - OBJECTIVE: Preoperative nasal continuous positive airway pressure(NCPAP) treatment was used as a substitute for protective tracheostomy before UPPP surgery in 42 patients with severe obstructive sleep apnea syndrome(OSAS) from 1994 to 1996. METHODS: Fifty-two patients operated on three years before this period served as control group. All cases were diagnosed as severe OSAS by polysomnography (PSC). RESULTS: There were no statistical difference in main parameters between two groups. Seven cases in the control group underwent protective tracheostomy whose apnea and hypopnea index(AHI) was 28.4-83.5 and lowest saturation oxygen (SaO2) were 7%-32%. One patient underwent emergency tracheostomy due to life-threatening situation during uvulopalatopharyngoplasty (UPPP). There were 9 similarly severe cases who did not undergo protective tracheostomy in NCPAP treatment group. All patients underwent preoperative 5-20 day NCPAP treatment in treatment group. The average positive pressure was 1.16 kPa(11.85 cmH2O). There were significant difference in main parameters between the two groups. The AHI, SaO2 and sleep structure were evidently improved. UPPP was safely performed without protective tracheostomy and any complications in all patients with NCPAP treatment. CONCLUSION: This result implys that NCPAP treatment could be used as a substitute for protective tracheostomy before UPPP in severe OSAS. PMID- 11498853 TI - [Advances in the cochlear implantation]. PMID- 11498854 TI - [Screening for the 1555G mutation in mitochondrial DNA in pedigrees with aminoglycoside antibiotic induced deafness]. AB - OBJECTIVE: To identify the relationship between the 1555G mutation in mitochondrial DNA and aminoglycoside antibiotic induced deafness and provide theoretical evidence for establishing diagnostic method. METHODS: Blood samples were obtained from three pedigrees with aminoglycoside antibiotic induced deafness. DNA was extracted from the isolated leukocytes. The mitochondrial DNA fragments were amplified by PCR, 1555G mutation was detected by Alw26 I restriction endonuclease digestion, allele-specific oligonucleotide hybridization and DNA sequencing. RESULTS: Seven individuals from pedigrees A and C carried homoplasmic 1555G mutation, six individuals from pedigree B did not have 1555G mutation. CONCLUSION: 1555G mutation is not the only pathologic mitochondrial DNA mutation associated with aminoglycoside antibiotic induced deafness. This is the first finding of aminoglycoside antibiotic induced deafness pedigree without 1555G mutation. PMID- 11498855 TI - [Study on nitric oxide in isolated cochlear neuron of chicken with chemically modified carbon fiber microelectrode]. AB - OBJECTIVE: Nitric oxide(NO) is involved in neural signaling that is important to the physiologic and pathophysiologic activities of cochlea and vestibule. This study is to compare physiologic function of mammalian cochlea with that of chicken cochlea. METHODS: Using chemically modified carbon fiber microelectrode to measure qualitatively NO which was released from isolated cochlear neuron cluster of chicken in response to certain agonists, such as ATP, L-Arg and ACh. The presence of NO release is indicated by elevation of amplitude and lasting time of microelectric current. RESULTS: The results demonstrated that No releases stimulated by ACh and L-Arg were similar in amplitude but the lasting time stimulated by ACh was shorter than that of L-Aeg. Response to ATP, however, differred apparently in a manner which the amplitude was larger than that of ACh and L-Arg, whereas the lasting time approximated to that of L-Arg but longer than that of ACh. Before application of agonists, preapplication of L-NNA, a NOS antagonist no current changes could be monitored. CONCLUSION: These data suggested that chicken cochlear ganglion can synthesize and release NO. The effective mechanism of the agonists and possible role of NO in the cochlea is discussed. PMID- 11498856 TI - [The effect of calcium ion in perilymphatic fluid on guinea pig's distortion product otoacoustic emissions]. AB - OBJECTIVE: To study the mechanism of the production of distortion product otoacoustic emissions (DPOAE) by electromobility of out hair cell. METHODS: The changes of nine gninea pig's DPOAE by means of perfusion of artificial perilymphatic fluid with and without calcium ion were observed. RESULTS: The mean value of DPOAE decreased from 41.3 dB to 13.7 dB after perfusion of fluid devoid of calcium ion, it revived to 36.7 dB after reperfusion with normal perilymphatic fluid. The result supports Pickle's muscle theory for DPOAE's mobile mechanism and does not support Brownell's electromobility theory. CONCLUSION: A normal concentration of calcium ion in perilymphatic fluid is necessary for outer hair cell to produce DPOAE. PMID- 11498857 TI - [Quantitative analysis of vestibulo-ocular reflex and optokinetic reflex electronystagmography in patients with unilateral loss of vestibular function]. AB - OBJECTIVE: To study changes of the vestibular function in the process of vestibular compensation. METHODS: The electronystagmography recording of spontaneous nystagmus(SN) with tests for vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) were performed in 105 patients with unilateral loss of vestibular function, the results were analysed quantitatively. RESULTS: (1) Twenty-one patients with courses from 3 days to 6 years presented a SN directed towards the intact side, with an average slow phase velocity (ASPV) of 7.12-12.07 degrees/s. (2) The intensity of VOR nystagmus as a response to the warm-cold water stimulation to the intact ear was lower than that of normal subjects. (3) By 0.167 Hz, 90 degrees sinusional oscillation stimulation, the ASPVs of the patients' VOR nystagmus directed towards the intact and affected sides were lower than that of the normal. A predominance of VOR nystagmus rotation towards the intact side was found in patients with courses < or = 5 years. There was no predominance of VOR nystagmus found in patients with courses > 5 years. (4) There was no significant difference between the optokinetic nystagmus (OKN) intensity of the normal persons and that of patients with lesions outside the cerebellopontine angle and with tumors in cerebellopontine angle which was < 2.5 cm in diameter. The OKN intensity of patients with tumors in cerebellopotine angle with diameters > or = 2.5 cm was lower than that of normal subjects. A predominance of OKN towards the intactside was found in these patients. CONCLUSION: The vestibular function is in inhibitory and the vestibular system becomes rebalanced due to this inhibition in the process of vestibular compensation. PMID- 11498858 TI - [Jugular bulb anomaly]. AB - OBJECTIVE: To differentiate and treat correctly the jugular bulb anomalies, as well as to avoid the unnecessarily enlarged operations. METHODS: Four cases of jugular bulb anunalies were reported and analysed. RESULTS: Two of 4 cases were misdiognosed as jugular glomus tumors; One was considered to be jugular bulb anomaly, but was done with an enlarged operation; another one was still accompanied with benign intracranial hypertension syndrome, was inappropriately treated. CONCLUSION: The jugular bulb anomaly was often misdiagnosed, therefore the anomaly should be differentiated from the jugular glomus tumor carefully. PMID- 11498859 TI - [Skull base operations via pre-auricular temporo-sphenoidal approach and reconstructions of their defects]. AB - OBJECTIVE: To study the surgical approach for cranial base tumors removal. METHODS: Ten cases of cranial base tumors were thoroughly resected via pre auriculal temporo-sphenoidal approach. Among them, 9 cases with defects of cranial base or nasopharynx were reconstructed with the temporal muscle and fascia in the same operative field. RESULTS: Patients were followed up from 21 months to 7 years. In 2 cases with recurrent cranial base tumors 1 died of recurrence, 1 lived with the tumor. Eight cases achieved good clinical results without severe complications except 1, who got necrosis of temporal muscle. CONCLUSION: The preauricular temporo-sphenoidal approach not only provides an excellent exposure to the compartments of the skull base with a limited injury to the normal tissues, but also offers a convenient way in reconstructing defects with temporal muscle in the same operative field. The incision scar is hidden and the eosmetic affect small. It has been shown that the approach can offer some advantages in surgical treatment of cranial base tumors selectively. PMID- 11498860 TI - [Change of cell proliferation after the denervation laryngeal adducent muscles in dogs]. AB - OBJECTIVE: To investigate the changes of cell proliferation state after denervation of laryngeal adducent muscles in dog. METHODS: The muscles were obtained from 8 dogs with recurrent lanyrgeal nerves injured. The cross sectional areas of muscle cells were measured by VIDS Image Analysis System in HE stained sections. A quantitative collagen/muscle ratio was measured by TJTY-300 image analysis system in Van Gieson stained sections. The percentages of S phase cell nucleus were calculated by image cytometry/DNA quantitative analysis with CAS 200 System. RESULTS: It showed that the cross sectional area of muscle cells decreased gradually after denervation (out-standing in 6 months after denervation) and collagen/muscle ratio increased significantly with time. The percentage of S phase nucleus was high at 5 months and then decreased. CONCLUSION: The proliferation of thyroarytenoid muscle and lateral cricoarytenoid muscle cell of dog remained at high level in 5 months after denervation and then slowed down and atrophy of muscles occurred 6 months after denervation. This suggests that nerve repair should be performed early. Examination of cell proliferation might indicate if the denervation of cells irreversible. It is helpful in choosing proper surgery and estimating the effect of nerve repair. PMID- 11498861 TI - [Application of three demensional reconstruction of CT in lateral cranial base surgery]. AB - OBJECTIVE: To study the characteristics of 3D-CT of the lateral cranial base and its application in lateral cranial base surgery. METHODS: Five normal adult subjects and 10 with lateral cranial base lesions were scanned with Philips Tomoscan SR 7000 CT scanner. The scanning ranges extended from the top to the bottom of the temporal bone, including whole lesions, with bone window algorithm and contiguous serial 1.5 mm thin-cut scans in succession in axial plane. The 3D CT reconstruction was processed in the Easy Viewing CT/MRI workstation. RESULTS: The 3D-CT reconstruction could clearly delineate the foraminan, fissures and important bony marks. The ranges of the lesions, the relationships between the lesions and the structures around them were demonstrated more clearly than 2D-CT. It was of important significance in differential diagnosis for jugular foramen lesions. 3D-CT could be used to imitate operation and design operative plan. CONCLUSION: 3D-CT was of important value in diagnosis of lateral cranial base tumors, and could be used to imitate operation and design operative plan. PMID- 11498862 TI - [Expression of ras, p53, proliferating cell nuclear antigen in squamous cell carcinoma of maxillary sinus and their significance]. AB - OBJECTIVE: To determine the expression of ras, p53 and proliferating cell nuclear antigen (PCNA) in squamous cell carcinoma(SCC), inverting papilloma(IP), inflammatory mucosa(IM) of maxillary sinus and analyze the correlation between their expression and the clinicopathological characters and prognosis of SCC of maxillary sinus(SCCMS). METHODS: Expression of ras, p53 and PCNA was determined in 40 cases of SCC, 20 cases of IP and 10 cases of IM of maxillary sinus with immunohistochemical technique. RESULTS: Expression of ras was correlated with differentiation of SCCMS, it was more frequent in well differentiated SCC than in poorly differentiated SCC. Expression of p53 was only present in SCCMS, there was not expression of p53 in IP and IM of maxillary sinus. The patients with high PCNA index SCCMS had a poorer prognosis than those with low PCNA index in SCCMS. CONCLUSION: p53 protein may be a diagnostic marker in SCCMS. Measurement of PCNA index may be helpful in estimating prognosis and directing clinical treatment in SCCMS. PMID- 11498863 TI - [Endoscopic transsphenoidal approach in surgical treatment of pituitary adenomas]. AB - OBJECTIVE: To recognize the indications and the value of endoscopic transsphenoidal approach in surgical treatment of pituitary adenomas. METHODS: The management of pituitary adenomas in 24 patients was reviewed. RESULTS: Endoscopic transnasal transspheniodal approach was used successfully in 24 cases and endoscopic transseptal transsphenoidal approach was used successfully in 4 cases, total tumor resection in 23 cases and subtotal resection in 1 case were achieved. CONCLUSION: This new technique offered a simple and rapid access to the sella turcica, and also, direct visualization which improve the surgeon's ability to resect tumors, it is a safe, valuable and efficient procedure for removing pituitary adenomas. The indications and management of the complications of endoscopic pituitary surgery were discussed. PMID- 11498864 TI - [Reversal of multidrug resistance by cyproheptadine in KBV200 cells]. AB - The cyproheptadine (CYP) reversal of multidrug resistance (MDR) and its mechanism in KBV200 cell line were studies. MTT assay showed that CYP 15.0 mumol.L-1 could reverse vincristine, adriamycin (ADR) and etoposide resistance in KBV200 cells by a factor of 5.5, 2.0 and 1.9, respectively. CYP appeared to have no influence on the cytotoxicity of 5-fluorouracil (5-FU) and melphalan (MEL) in the cells. These results indicate that CYP is a MDR reversing agent. CYP 15.0 mumol.L-1 increased the ADR accumulation in KBV200 cells from 0.68 +/- 0.03 microgram/10(6) cells to 1.36 +/- 0.08 micrograms/10(6) cells (P < 0.01). CYP 15.0 mumol.L-1 was shown to obviously increase rhodamine 123 (R123) accumulation in and decrease its efflux from the cells. There was no change in PGP dying intensity under immunocytochemical assay and in mdr1 RNA level through slot blot analysis in the KBV200 cells exposed continuously to CYP 15.0 mumol.L-1 for 72 h. These results suggest that CYP acts by inhibiting the pumping function of PGP. PMID- 11498865 TI - [Effects of tachykinin receptor antagonists on allergic asthma in guinea pigs]. AB - In conscious sensitized guinea pigs, CP-96345 (2.06 mumol.kg-1, i.p.), a specific antagonist for tachykinin NK-1 receptors, SR-48968 (1.66 mumol.kg-1, i.p.), an NK 2 receptor antagonist, and the combination of both agents decreased the wheezing percentage and the mortality from anaphylactic shock induced by 0.25% ovalbumin (OA, for 0.5 or 2 min) aerosol inhalation. In the anesthetized guinea pigs, SR 48968 attenuated OA (5 mg.kg-1, i.v.)-induced bronchoconstriction, while CP-96345 inhibited OA-induced Evans blue extravasation in bronchi and intrapulmonary airways. In the isolated tracheal and bronchial smooth muscle preparations of guinea pigs, SR-48968 concentration-dependently inhibited OA (10 micrograms.ml-1) induced contraction both in trachea and in bronchi, while CP-96345 only attenuated the contraction of bronchi. Pretreatment with capsaicin, a depleting agent of sensory neuropeptides from sensory nerve C-fibers, attenuated the OA induced contractions both in trachea and in bronchi. The results indicate that (1) tachykinins in the airways are involved in the pathogenesis of allergic asthma; (2) tachykinin receptor antagonists have inhibitory effects on the allergic asthmatic responses, which is at least partly through the inhibition of antigen-induced contraction of airway smooth muscles (NK-2 receptor effect) and airway microvascular leakage (NK-1 receptor effect). PMID- 11498866 TI - [Protective action of corynoline, acetylcorynoline and protopine against experimental liver injury in mice]. AB - Oral administration of two doses of corynoline, acetylcorynoline or protopine at 50 and 100 mg.kg-1 in an interval of 8 to 24 h before i.p. injection of CCl4, acetaminophen or thioacetamide significantly impeded the elevation of serum transaminase (SGPT) and liver damage in mice. The three compounds were found to inhibit CCl4-induced microsomal lipid peroxidation and CCl4 conversing to carbon monoxide in liver microsomes in vitro. Of these compounds, acetylcorynoline was shown to be more potent than corynoline and protopine. In addition, all the three compounds exhibited biphasic effects on the hepatic cytochrome P450, i.e. inhibition followed by induction, in mice. PMID- 11498867 TI - [Effect of HH07A, A derivative of hainanensine, on DNA polymerase I]. AB - The effect of HH07A on DNA polymerase I in a cell free system was studied. The results showed that HH07A strongly inhibited the DNA synthesis catalyzed by DNA polymerase I in a dose dependent manner. Further results indicated that the DNA synthesis decreased significantly after DNA polymerase I was preincubated with HH07A. However, no change was found when DNA template was preincubated with HH07A. These results suggest that the inhibitory action of HH07A appeared to be on the enzyme molecules rather than on the DNA template. PMID- 11498868 TI - [Protective effects of D-3-N-butylphthalide and L-3-N-butylphthalide on neuronal damage induced by KCl and NMDA in cultured rat cortical neurons]. AB - The protective effects of l-3-n-butylphthalide (l-NBP) and d-3-n-butylphthalide (d-NBP) on KCl (20 mmol.L-1)- or NMDA (N-methyl-D-aspartate, 30 mumol.L-1) induced cytotoxicity in primary cultured rat cortical neurons were studied. Intracellular LDH release, percentage of cell death and cellular morphological changes were used to evaluate the effect of drugs. l-NBP (1-100 mumol.L-1) and d NBP (1-100 mumol.L-1), but not nimodipine (1-100 mumol.L-1), were shown to dose dependently inhibit LDH release induced by NMDA (30 mumol.L-1) in cultured rat cortical neurons with IC50 values of 4.89 mumol.L-1 and 13.52 mumol.L-1, respectively. The percent cell death was reduced with IC50 values of 44.37 and 49.78 mumol.L-1, and the cellular morphology improved. The effect seemed to be the same as that of equal concentration of NAME(NG-nitro-L-arginine methyl ester). In addition, l-NBP(10 mumol.L-1), d-NBP (10 mumol.L-1) and nimodipine(10 mumol.L-1) also produced significant inhibition on intracellular LDH release and decrease in percent cell death induced by KCl(20 mmol.L-1) in cultured neurons. The potencies of l-NBP and d-NBP were similar to that of equal dose of nimodipine. These data suggest that l-NBP and d-NBP can remarkably protect cultured neurons against the damage induced by KCl and NMDA. PMID- 11498869 TI - [Synthesis and antibacterial activity of tricycic fluoroquinolones]. AB - Twelve new analogues of new tricyclic rufloxacin were prepared and their MIC were evaluated against thirteen kinds of bacteria. As a result of these studies: the polarity of C10-side chain was found to exert greater positive effect on G- than on G+ bacteria. PMID- 11498870 TI - [Structure determination of new antifungal antibiotics, polaramycins A and B]. AB - Antifungal antibiotics, polaramycins A and B have been isolated from the culture of Streptomyces hygroscopicus LP-93. Polaramycins A and B are 36-membered polyol macrolide antibiotics containing hemiacetal monoester of malonic acid and guanidyl groups. Based on spectral evidence including UV, IR, FABMS, 1HNMR, 13CNMR, DEPT, 1H-1H COSY, HETCOR, HMBC, the structure of polaramycins A and B have been assigned as shown in Fig 1 (A, R = H; B, R = CH3). PMID- 11498871 TI - [Studies on the iridoid glucosides of the root of Lamiophlomis rotata (benth.) kudo, a medicinal plant in Xi zang (Tibet)]. AB - Lamiophlomis rotata (Labiatae) is a Chinese folk medicinal plant in Xi-zang (Tibet), which has effects of promoting blood circulation to remove blood stasis, subduing swelling and alleviating pain. Four iridoid glucosides were isolated from the root of the species. On the basis of IR, UV, NMR, MS spectral data and chemical methods, their structures were identified as: 8-O-acetylshanzhiside methyl ester(I), 6-O-acetylshanzhiside methyl ester(II), penstemoside(III) and 7, 8-dehydropenstemoside(IV). The last one is a new compound. PMID- 11498872 TI - [Studies on chemical constituents of Polygala arillata buch-ham]. AB - Two new compounds were isolated from the roots of Polygala arillata Buch-Ham. On the basis of chemical reactions and spectral (UV, IR, MS, 1HNMR, DIFNOE, 13CNMR) analysis, they were identified as 1,3-dihydroxy-2-methoxyxanthone(I) and 7 hydroxy-1-methoxy-2,3-methylenedioxyxanthone(II). Pharmacological study indicated that I and II have inhibitory effect on aldose reductase activity. PMID- 11498873 TI - [Isolation and identification of four new tetracyclic-atxanes from Taxus yunnanensis]. AB - Four new minor toxoids, named as taxayuntin A, B, C and D, have been isolated from the ethanolic extract of the bark of Taxus yunnanensis Cheng et L. K. Fu. Their structures were identified as the new taxane diterpenoids with 5/7/6/4 membered ring system by spectral means and 2D NMR shift correlation experiments. PMID- 11498874 TI - [The sesquiterpenoids from Celastrus flagellaris]. AB - A new sesquiterpenoid (1) named as celastrine B along with seven known compounds 1 alpha, 6 beta, 13-triacetoxy-9 beta-benzoyloxy-beta-dihydroagarofuran(2), triptogelin G-1(3), 1 alpha, 6 beta-diacetoxy-9 beta-benzoyloxy-beta dihydroagarofuran (4), triptogelin F-2 (5), 1 alpha, 2 alpha-diacetoxy-9 beta cinnamoyloxy-beta-dihydroagarofuran (6), celaforlin B-3 (7), 1 alpha, 6 beta diacetoxy-8 alpha-cinnamoyloxy-9 alpha-benzoyloxy-beta-dihydroagarofuran(8) were isolated for the first time from the seed oil of Celastrus flagellaris Rupr. and their structures elucidated by UV, IR, MS and 1HNMR, 13CNMR, DEPT, 1H-1H COSY, 13C-1H COSY, NOESY and HMBC. PMID- 11498875 TI - [Polarographic behavior of latamoxef sodium and its trace determination]. AB - In the medium of HCl-KCl (pH 1-2) by means of cyclic voltammetry and differential pulse polarography (DPP), the mechanism of the electrode reaction for latamoxef sodium (Shiomarin) at static mercury drop electrode (SMDE) and hanging mercury drop electrode (HMDE) has been proposed. A sensitive method for the determination of nanomolar concentration of latamoxef sodium by DPP was developed. The peak potential was -0.62 V (vs Ag/AgCl). The linear range was from 1 x 10(-8) to 8 x 10(-4) mol.L-1, the detection limit was ca. 4 x 10(-9) mol.L-1(tac = 90 s). The method was applied to the determination of latamoxef sodium in injection with satisfactory results. The possibility for the direct determination of latamoxef sodium in mimic urine sample was also studied. PMID- 11498876 TI - [A systematic screening and identification method for 29 central nervous system drugs in body fluid by high performance capillary electrophoresis]. AB - A systematic screening method has been developed for the detection of 29 central nervous system (CNS) drugs in human plasma, urine and gastric juice by high performance capillary electrophoresis (HPCE). The first step is sample preparation. The patient's or normal human plasma (0.5 ml) spiked with CNS drugs was extracted with 2 x 4 ml dichloromethane, while 2 ml of patient's or spiked urine was extracted with 2 x 6 ml chloroform. The combined extract from plasma or urine was evaporated to dryness in a rotation evaporator at 35 degrees C. The residue was dissolved in 100 microliters methanol and subsequently 400 microliters of redistilled water was added. The patient gastric juice (3 ml) was centrifuged at 2,000 r.min-1 for 5 min. The supernatant was filtered through 0.45 micron microporous membrane for injection onto capillary columns. The second step was to perform CZE separation in acidic buffer composed of 30 mmol.L 1(NH4)3PO4(pH 2.50) and 10% acetonitrile (condition A). Most of the benzodiazepines (diazepam, nitrazepam, chlordiazepoxide, flurazepam, extazolam, alprazolam) and methaqualone were baseline separated and detected at 5-13 min, while thiodiphenylamines showed group peaks at 3-5 min and barbiturates migrate with electroosmotic fluid (EOF) together. The third step is to separate the drugs in basic buffer constituted of 70 mmol.L-1 Na2HPO4(pH 8.60) and 30% acetonitrile (condition B). The thiodiphenylamines and some other basic drugs could be well separated, which include thihexyphenidyl, imipramine, amitriptyline, diphenhydramine, chlorpromazine, doxepin, chlorprothixene, promethazine and flurazepam, while the rest of the CNS drugs did not interfere with the separation. The last step was to separate the drugs by micellar electrokinetic chromatography (MEKC) in such a buffer as 70 mmol.L-1 SDS plus 15 mmol.L-1 Na2HPO4 (pH 7.55) and 5% methanol (condition C). Barbiturates (barbital, phenobarbital, methylphenobarbital, amobarbital, thiopental, pentobarbital, secobarbital) and some hydrophobic drugs (glutethimide, alprazolam, clonazepam, carbamazepine, trifluoperazine, oxazepam) could be well separated. These drugs might be identified by both the relative migration time (rtm = tdrug/tEOF) and the ratios of peak heights (rh) monitored at different wavelength, since the ratios are characteristic of the spectrum of a drug. This method has been used in several real clinical samples of intoxication. For example, perphenazine and doxepin were detected in the gastric juice and phenobarbital in blood and gastric juice of an intoxicated patient. PMID- 11498877 TI - [A preliminary study on the identification of crude snake drugs by molecular genetic markers]. AB - DNA was amplified from 11 samples of crude snake drugs by 2 random primers. "Wushaoshe" (Zaocys dhumnades) and its substitutes, Elaphe taeniurus, E. rufodorsata, E. carinata and Sinonatrix annularis, and "Jinqian Baihyashe" (Bungarus multicinctus) and its adulterants, Dinodon rufozonatum and Sinonatrix annularis, can be distinguished by electrophoresis pattern of the amplified products. The results showed that the molecular genetic marker technique can be used as a new method for the identification of crude snake drugs. PMID- 11498878 TI - [Progress in the chemistry of plant cyclopeptides]. PMID- 11498879 TI - [Thermal influence on intracellular calcium concentration of isolated vestibular hair cells in guinea pig]. AB - OBJECTIVE: To further understand the mechanism of caloric test. METHODS: Vestibular hair cells(VHC) were isolated from guinea pig crista ampullaris by enzymatic and mechanical methods and the variations of intracellular free calcium ion concentration ([Ca2+]i) with changes of the extracellular medium temperature were observed. The [Ca2+]i of VHC was examined using laser scanning confocal microscopy (LSCM) and the Ca2+ sensitive dye Fluo-3. RESULTS: Were as follows: 1. There was an increase in [Ca2+]i of 81.8% (18/22) type I VHC and 7/12 type II VHC with the heat stimulation and a decrease in [Ca2+]i with temperature decrease. The magnitude of variation in [Ca2+] i in type I VHC was larger than that in type II VHC. 2. There was no significant change in [Ca2+]i of outer hair cells (OHC) with the heat stimulation. CONCLUSION: There is a direct thermal influence on vestibular sensory cells and this influence accompany with the variation of [Ca2+]i in VHC. PMID- 11498880 TI - [Discernment of the sensitized inner ear by peripheral immunocompetent lymphocytes]. AB - OBJECTIVE: To understand the role of peripheral immunocompetent lymphocytes during an immune response in the inner ear. METHODS: Sensitized lymphocytes from peripheral blood, neck lymph nodes and spleen from three groups of donor animals (group A was immunized intradermally with KLH and both inner ears were challenged with KLH; group B received only KLH intradermally without an inner ear challenge, and groups C were naive animals) were labelled with 51Cr and injected intravenously into recipients (all of which were immunized intradermally with right inner ears challenged with KLH). Twenty to twenty-four hours later, the temporal bones of recipients were assayed for radioactivity to detect the infiltration of labelled cells. RESULTS: In group A and B, the right temporal bones showed higher radioactivity than the left temporal bones (P < 0.05). The highest labelling was detected in the right temporal bones of group A. The lowest migration of 51Cr-labelled cells to the inner ear was found in group C animals. The highest ratios between right (challenged) and left (unchallenged) were found in group A animals compared with those of group B and group C. The value was significantly higher in group A than in group B and C (P < 0.05). CONCLUSION: The results showed that the inner ear comes under the immunosurveillance of the peripheral circulation in response to antigenic stimulation. PMID- 11498881 TI - [Inhibitive effect of bcl-2 antisense oligodeoxynucleotide on Bcl-2 protein synthesis and cell proliferation in human laryngocarcinoma Hep-2 cells]. AB - OBJECTIVES: To evaluate the role of bcl-2 antisense oligodeoxynucleotide in the gene treatment of human laryngocarcinoma. METHODS: An antisense oligodeoxynucleotide complementary to the initiation coden and the next four codens of bcl-2 mRNA was synthesized with an automatic DNA synthesizer. It was used to treat cultured Hep-2 cells to observe its effect on cell proliferation. In situ hybridization method and immunohistochemical technology were adopted to detect bcl-2 mRNA and protein and explore the effect of bcl-2 antisense oligodeoxynucleotide fragment on bcl-2 gene expression and protein synthesis and inhibition of cell proliferation. RESULTS: Bcl-2 antisense oligodeoxynucleotide had an obvious effect on bcl-2 mRNA, but inhibited Bcl-2 protein synthesis significantly, the inhibitive rate had positive correlation with concentration of antisense oligodeoxynucleotide and time of action. Twenty mumol/L of antisense oligodeoxynucleotide could inhibit cell proliferation effectively. CONCLUSIONS: It is suggested that bcl-2 antisense oligodeoxynucleotide might specifically inhibit bcl-2 protein synthesis and cell proliferation in Hep-2 cells at translation level. PMID- 11498882 TI - [Surgery of advanced clival tumors]. AB - OBJECTIVE: To investigate the effects of surgical procedures on tumor 27 patients with advanced clival tumors during the last 9 years. METHODS: Four different kinds of surgical approaches were selected and used according to preoperative clinical and radiographic findings, and the complications and the long-term survival rates were evaluated after the surgical procedures. RESULTS: All of the 8 patients with benign clival tumors (6 cases with chordomas and 2 with meningiomas) had over 2-8 years survival and no severe complications after the surgical procedures and adjuvant radiotherapy. Fourteen of the 19 patients with malignant tumors had over 2-6 years survival (a survival rate of 73.7%) after the surgical procedures, the adjuvant chemotherapy and radiotherapy. CONCLUSION: Surgery of the clivus appears to be worthwhile procedure for improving outcome of the patients with advanced clival tumors. PMID- 11498883 TI - [Long term results of Majer-Piquet's operation in the treatment of laryngeal carcinoma]. AB - OBJECTIVE: To investigate the long term results of Majer-Piquet's operation in the treatment of glottic type of laryngeal carcinoma. METHODS: A series of 32 cases operated on between October 1990 and February 1995 were analysed, of whom 21 were T1N0M0, 3 were T1N1M0, 5 were T2N0M0 and 1 each for T2N1M0, T3N0M0, T3N1M0. RESULTS: In T1 cases, 2, 3 and 5 years survival rate were 95.8, 93.7 and 90.9% respectively, while in T2 cases, 2, 3 and 5 years survival rate were 100%, 83.3% and 60% respectively. Decannulation rate was 96.9%. All patients could finally take food by mouth without inspiration, and could speak as soon as decannulated. By modifing the operative technique, the cases operated on after 1994 got relatively better results in phonation. CONCLUSION: Majer-Piquet's operation not only is effective in the treatment of T1, T2 and some T3 glottic type of laryngeal carcinomas, but also can satisfactorily reserve laryngeal function and improve the quality of patient's life. PMID- 11498884 TI - [The experimental study of the facial nerve regeneration in silicone chamber: the influence of nerve growth factor]. AB - OBJECTIVE: To elucidate the role of NGF in the regeneration of facial nerve. METHODS: The superior buccal division of facial nerve of adult New Zealand rabbit was transected and a nerve growth chamber created. The chamber of the experimental side was filled with NGF/normal saline and that of the control side with normal saline alone. Four and eight weeks after operation, the regenerated nerves in the chambers were dissected for histological studies. RESULTS: Four weeks after operation, the average thickness of myelin sheath and the average number of myelinated axons were 0.779 +/- 0.475 micron, 2.024 +/- 1.999 (n = 11) in experimental group and 0.413 +/- 0.132 micron, 368 +/- 171 (n = 8) in control sides respectively. There was significant difference between the experimental sides and control sides (P < 0.05). Eight weeks after operation, the regenerated nerve appeared more mature. There were significant difference in the average diameters, the thickness of myelin sheath and the number of myelinated axons between the experimental and control sides (P < 0.05). CONCLUSION: NGF within a silicone chamber enhanced facial nerve regeneration in New Zealand rabbits. PMID- 11498885 TI - [Quantitation of nerve fibers in habenula perforata in chinchilla]. AB - OBJECTIVE: To examine the number of nerve fibers in the habenula perforata of the cochlea in normal and carboplatin-treated chinchillas. METHODS: Three months following administration of carboplatin (100 mg/kg x 2), six carboplatin-treated animals, together with 6 controls, were sacrificed, prepared for semi-thin sectioning of the cochlea. The sections were stained with toluidine blue and basic fuchsin, examined under a light microscope and photographed. RESULTS: In normal chinchillas, the numbers of the nerve fibers in each habenula opening varied across the cochlea, with 93.7 +/- 12.2(mean +/- s) fibers at the hook, 181.5 +/- 30.4 at the upper of basal turn, 129.7 +/- 27.8 at the second turn and 75.8 +/- 13.2 at the apical turn. In cochlear devoid of type I ganglion cells and inner hair cells as a result of carboplatin treatment, only one-seventh of the fiber population remained. CONCLUSION: Approximately six-seventh of the nerve fibers in the habenula are type I afferent axons that innervate the inner hair cells. The remaining fibers are type II afferents and surviving efferents. PMID- 11498886 TI - [Effect of substance P on the short-circuit current of nasal mucosal epithelium]. AB - OBJECTIVE: To observe the changes of epithelial apical membrane short-circuit current (Isc) of antigen-sensitized rat nasal mucosa with the stimulation of Substances P (SP). METHODS: The epithelial apical membrane Isc were measured with Ussing chamber technique, and and we observed the blockage to the effect of SP by neurokinin receptor antagonist Cp96345, histamine H1 receptor antagonist pyrilamine, H2 antagonist ranitidine and neurotoxin respectively. RESULTS: The Isc increased significantly with the stimulation of SP. The effect of SP on Isc could be blocked significantly by pretreatment with four substances. CONCLUSION: SP releasing from nervous endings plays the role on elicit a series of pathological changes like enhanced Isc, epithelial permeabilities, etc. PMID- 11498887 TI - [Radioimmunological image in tumor bearing nude mice with monoclonal antibody against human laryngeal carcinoma]. AB - OBJECTIVE: To investigate the specific autoradiographic diagnosis of laryngeal carcinoma and its metastasis. METHODS: Three strains of the mixed monoclonal antibodies against human laryngeal carcinoma (McAbLC9, 11, 12) labelled with 131I were injected(intraperitoneally) into the nude mice bearing squamous cell carcinoma. The tumor autoradiography was done by single photo-emission computed tomography (ECT). The percent injected dose/g of tumor were measured, and the radioactivity ratio of tumor to non-tumor tissues(T/NT) were calculated. RESULTS: The 131I-McAbLC 9, 11, 12 can accumulate in the area of the laryngeal carcinoma within limited time, and demonstrate a satisfactory tumor imaging at 96 h. In experimental groups: The %ID/g of tumor tissues (mean +/- s, n = 6) was 18.8 +/- 0.61; The %ID/g of tumor tissues in the control group was 4.46 +/- 0.82 (mean +/- s, n = 6) at 96 h. The difference was statistically significant (P < 0.01, t = 34.97). CONCLUSION: The results suggest that 131I McAbLC9, 11, 12 is specific and sensitive in recognising human laryngeal carcinoma cell, and the area and size of tumor can be easily detected by ECT. It is feasible for localization of tumor and its metastasis. PMID- 11498889 TI - [Study on the methodology of potassium channel expression in Xenopus oocytes by messenger RNA from rat cochlear nucleus]. AB - OBJECTIVE: To study the properties of voltage-dependent ion channels expressed in Xenopus oocytes by mRNA from rat cochlear nucleus and to lay a methodological foundation for studying the changes of ion channels in auditory centre with sensorineural hearing loss. METHODS: Poly (A)+ mRNAs isolated from the rat cochlear nucleus by acid guanidinum thiocyanate-phenol-chloroform (AGPC) method and Promega's new magnesphere technique were microinjected into Xenopus laevis oocytes to express functional ion channels. Voltage clamp technique was used to record the current of voltage-dependent ion channels. RESULTS: A voltage dependent ion channel was detected from oocytes injected with rat cochlear nucleus mRNA. The maximum amplitude of the outward current was 364 +/- 42 nA (n = 16). It was identified as transient outward potassium channel. CONCLUSION: The model set up by the test may play an important role in studying the physiology, pathology and pharmacology of auditory centre. PMID- 11498888 TI - [Experimental studies on antitumor activity of new recombinant human tumour necrosis factor in nude mice bearing nasopharyngeal carcinoma]. AB - OBJECTIVE: Nude mice model bearing nasopharyngeal carcinoma were established using nasopharyngeal squamous carcinoma cell line (CNE-2). METHODS: New recombinant human tumor necrosis factor (NrhTNF-alpha) and carboplatin were administered locally or systemically, single or in combination. The microstructural and ultra-micro-structural changes of carcinoma after administration of NrhTNF-alpha were observed under light and electron microscopes. RESULTS AND CONCLUSION: 1. After treatment, the tumor showed hemorrhage, necrosis, regression, the survival time was prolonged. 2. Locally administered NrhTNF-alpha had more significant antitumor effect than intravenously administered of NrhTNF-alpha. 3. There is a notable synergistic antitumor effect by the combination use of NrhTNF-alpha and carboplatin. 4. Microscopy showed that cell changes mostly took place in plasma especially in mitochondria and endoplosmic reticula. The mitochondria were highly bloated. Besides, some nuclei were condensed and fragmented, resulted in cell death. These studies provide experimental basis for biological treatment of nasopharyngeal carcinoma. PMID- 11498890 TI - [Diagnosis and surgical treatment of pituitary microadenoma]. AB - OBJECTIVE: Through the experiences of clinic picture and the radiologic characteristics of eleven cases of micropituitary adenomas, the early diagnosis and surgical indications were discussed. METHODS: Intensified CT and MRI were used to compare their significances in the diagnosis of micropituitary adenomas. Kaltai incision of the nares and transsphenoidal approach were adopted in these cases. RESULTS: The bulging of the upper border of the pituitary gland and the shifting position of the pituitary stalk observed in the intensified MRI are the pathognomonic signs for the diagnosis of micropituitary adenoma. The operation was stopped due to hemorrhage in one case. The average following up was 3.2 years past operative death in the second month in one case, operative failure in one case and loss of following up in one case. CONCLUSION: Strict selection of surgical indications must be done necessary after careful study of MRI. PMID- 11498891 TI - [Exploration of evaluating way of curative effect in transnasal endoscopic sinus surgery for treating chronic sinusitis]. AB - OBJECTIVE: To explore the evaluating way of curative effect in transnasal endoscopic sinus surgery for treating chronic sinusitis. METHODS: 725 cases who suffered from chronic sinusitis and undergone the endoscopic sinus surgery in 1991-1995 were analyzed according to the index that set up by ourselves. RESULTS: The difference of properties, limits and extents of pathological changes in sinuses affected directly the surgical result. CONCLUSION: It is very important to build up a staging system and a criterion of treating effect on chronic sinusitis for appraising properly the endoscopic sinus surgery. PMID- 11498892 TI - [Cerebrospinal aporrhinosis of nasal sinus operation]. AB - OBJECTIVE: To study the treatment of cerebrospinal aporrhinosis after nasal sinus operation, especially frontal, ethmoid tumor resection. METHODS: Fourteen cases of cerebrospinal aporrhinosis after nasal sinus operations were reported. RESULTS: Seven cases of cerebrospinal apporrhinosis were found during the operation. Two cases were repaired with tensor facia latae, 3 with mucosoperichondrium of nasal septum and EC otocerebral glue and 1 with TJ bone cement. Seven cases cerebrospinal aporrhinosis were found after the operations when intranasal packing was removed. They were cured by cerebral decompression and antibiotics, none complicated with intracranial infection. CONCLUSION: Careful examination should be taken daring the operation. If aporrhinosis was found, it should be repaired immediately. PMID- 11498893 TI - [The value of cephalometric analysis in diagnosis and treatment of obstructive sleep apnea syndrome]. AB - OBJECTIVES: To study the value of cephalometric analysis in the diagnosis, treatment option and evaluation of prognosis. METHODS: A detailed cephalometric analysis was performed using lateral X-ray films from 50 cases with obstructive sleep apnea syndrome (OSAS) and 30 age- and sex-matched controls. RESULTS: Statistical results showed that OSAS patients were different from controls in the following aspects: 1. Their uvula, soft palate and tongue were significantly enlarged. 2. The hyoid bone was displaced inferiorly and posteriorly. 3. The posterior airway space (palatopharyngeal plane) was narrow. 4. The hard palate was elongated. 5. The mandibular body was retropositioned. Fifty unselected consecutive OSAS patients(mean AHI 51.42 +/- 3.96, Medilog SAC 847, Oxford UK) underwent LAUPP using CO2 laser. Twenty-five cases were followed-up for 4-8 months. 10/25 had obtained a reduction in AHI > 50%, 15/25 had AI < 20 times. The success rate was compared with cephalomentric findings. CONCLUSION: Cephalometry has an important diagnostic and predictive value in the clinical investigation of patients suspected of having OSAS. PMID- 11498894 TI - [Immunocytochemical study of cholinergic innervation in the neurosensory epithelia of human vestibule]. AB - OBJECTIVE: To investigate the cholinergic innervation of the neurosensory epithelia of human vestibule. METHODS: A modified preembedding immunostaining technique for immunoelectronmicroscopy was applied to this study. A polyclonal antibody to choline acetyltransferase (ChAT) was used as the marker of cholinergic fibers. RESULTS: ChAT-immunoreactive products were restricted to the nerve fibers and terminals which were rich in synaptic vesicles. The ChAT immunoreactive fibers synaps with afferent chalice as well as with type II sensory hair cells. CONCLUSION: This study demonstrates that cholinergic fibers innervate the neurosensory epithelia of human vestible. The cholinergic fibers of human vestibular sensory epithelia belong to the vestibular efferent system. PMID- 11498895 TI - Effects of acupuncture on humoral immunologic function and trace elements in 20 cases of Behcet's disease. AB - Acupuncture therapy could normalize both increased light chain K value of IgM and lowered trace element Zn content in patients with Behcet's disease with statistically significant differences, suggesting that acupuncture therapy can elevate humoral immunologic function and improve metabolism of the trace element in these patients. PMID- 11498896 TI - Clinical study on treatment of senile vascular dementia by acupuncture. AB - 63 patients with senile vascular dementia were randomly divided into the treatment group (treated by acupuncture) and the control group (treated with piracetam). The authors observed the changes in the score of Hasegawa's dementia scale (HDS), p300, rheoencephalogram, topographic EEG, superoxide dismutase (SOD) activity in erythrocytes, and lipid peroxide (LPO) level in plasma before and after treatment. The statistical data showed that the total effective rate in the treatment group (80.6%) was significantly higher than that in the control group (25%), and the differences in the observed indexes before and after treatment were significant (P < 0.05 or P < 0.01) in the treatment group but not in the control group (P > 0.05), indicating that the acupuncture treatment was superior in immediate therapeutic effect on senile vascular dementia to drug treatment. PMID- 11498897 TI - Treatment of protrusion of the lumbar intervertebral disc by massotherapy. PMID- 11498898 TI - Observation on 105 cases of duodenal bulbar ulcer treated by combined therapy of catgut embedding and Chinese drugs. PMID- 11498899 TI - Catgut point-embedding therapy in treatment of 76 cases of ulcerative colitis. PMID- 11498900 TI - Treatment of obstinate diseases by acupuncture and cupping. PMID- 11498901 TI - An analysis of therapeutic effect of drug acupoint application in 209 cases of allergic asthma. AB - Both therapies of traditional crude herb moxibustion and drug acupoint application were used in 209 cases of allergic asthma to compare their long-term and short-term therapeutic effects and to analyze the relationship between clinic therapeutic effects of both therapies and differential types of the disease. The results showed that the short-term total effective rate in the group of drug acupoint application was higher than that in the group of traditional crude herb moxibustion, the therapeutic effects of drug acupoint application being closely related to differential types. Analysis also shows the best short-term therapeutic effect was in the type of heat in the lung while the poorest effect in the type of deficiency of the kidney-yang. PMID- 11498902 TI - Forty-seven cases of gonitis treated by a combined therapy of Chinese drugs and acupuncture. PMID- 11498903 TI - Treating principles and methods of traditional Chinese medicine in treatment of peripheral vascular diseases. PMID- 11498904 TI - Effect of Gan Fu Le on 24 cases of primary hepatocarcinoma. PMID- 11498905 TI - Effect of batroxobin on neuronal apoptosis during focal cerebral ischemia and reperfusion in rats. AB - We have found that Batroxobin plays a protective role in ischemic brain injury, which attracted us to investigate the effect of Batroxobin on apoptosis of neurons during cerebral ischemia and reperfusion. The apoptotic cells in ischemic rat brains at different reperfusion intervals were tested with method of TdT mediated dUTP-DIG nick end labeling (TUNEL) and the effect of Batroxobin on the apoptosis of neurons was studied in left middle cerebral artery (LMCA) occlusion and reperfusion in rat models (n = 18). The results showed that few scattered apoptosis cells were observed in right cerebral hemispheres after LMCA occlusion and reperfusion, and that a lot of apoptosis cells were found in left ischemic cortex and caudoputamen at 12 h reperfusion, and they reached peak at 24 h-48 h reperfusion. However, in the rats pretreated with Batroxobin, the number of apoptosis cells in left cerebral cortex and caudoputamen reduced significantly and the neuronal damage was much milder at 24 h reperfusion than that of saline treated rats. The results indicate that administration of Batroxobin may reduce the apoptosis of neurons induced by cerebral ischemia and reperfusion and afford significant cerebroprotection in the model of focal cerebral ischemia and reperfusion. PMID- 11498906 TI - Advances in TCM research of insulin resistance. PMID- 11498907 TI - Present situation in preventing and treating liver fibrosis with TCM drugs. AB - Considerable evidences have shown that the mechanism of TCM drugs for preventing and treating liver fibrosis is very complicated. TCM treatment can not only inhibit viral replication, ameliorate inflammation and promote blood circulation in the liver, and enhance regeneration of the hepatic cells, but also inhibit HSC proliferation, intra- and extracellular secretion, decrease the secretion of collagen and promote its degradation and re-absorption. However, most of the animal models are only suitable for studies of acute hepatitis. Establishment of cell lines suitable for studies of fibrosis is still at its initial stage. What we expect is that comprehensive clinical studies in TCM treatment of liver fibrosis will be carried out and evaluation of each datum given, both of which are of importance. PMID- 11498908 TI - A brief introduction to researches on treatment of hormone-dependent asthma with traditional Chinese medicine. PMID- 11498909 TI - Illustrative consultation of myelodysplastic syndrome. PMID- 11498910 TI - Treatment of nonspecific chronic prostatitis with Qian Lie Xian Yan Suppository suppository in 104 cases. AB - Qian Lie Xian Yan Suppository ([symbol: see text]) was rectally applied to treat 104 patients with nonspecific chronic prostatitis (damp-heat syndrome with blood stasis) in contrast to 30 patients treated with traditional Chinese drug Ye Ju Hua Shuan ([symbol: see text] Suppository of Flos Chrysanthemi Indici). The results show that chronic prostatitis is markedly improved by using Qian Lie Xian Yan suppository, with a short-term cure rate of 23.1% and a total effective rate of 84.6%, superior to that of the control group. The animal experiment indicates that Qian Lie Xian Yan suppository has better anti-inflammatory and analgesic effects, with an action of promoting blood circulation. PMID- 11498911 TI - Clinical study on treatment of chronic renal failure with shenshuailing. AB - The therapeutic effects of Shenshuailing Kou Fu Ye (SKFY [symbol: see text], the Oral Liquid for Renal Failure) and Shenshuailing Guan Chang Ye (SGCY [symbol: see text], the Enema for Renal Failure) were evaluated in treatment of chronic renal failure, with coateg aldehyde oxystarch as the controls. The changes in the clinical symptoms, serum creatinine, blood urea nitrogen and creatinine clearance rate were observed. The total effective rate in the former was 90.46%, and the latter 60.43%. PMID- 11498912 TI - The therapeutic effects of Sheng Jing Zhong Zi Tang in treating male infertility. AB - Eight-seven cases of male infertility due to spermatopathy were treated with Sheng Jing Zhong Zi Tang (SJZZT [symbol: see text]) for 1-3 courses. The total effective rate was 95.40% (83/87). 56.32% (49/87) of their spouses got pregnant and the semen quality was markedly improved (P < 0.01). The results showed that the decoction could dual-directionally regulate the levels of follicle stimulating hormone (FSH), prolan B luteinizing hormone (LH), testosterone (T), and cortisol (C). PMID- 11498913 TI - [Determination of acetylcholine and choline in microdialysates from rat brain by high performance liquid chromatography with electrochemical detection combined with a post-column immobilized enzyme reactor]. AB - In the present work, acetylcholine(ACh) and choline(Ch) in the microdialysates from three brain areas of anesthetized rats and from hippocampus and frontal cortex of freely moving rats were simultaneously measured by high performance liquid chromatography(HPLC) with electrochemical detection combined with a post column immobilized enzyme reactor(IMER). This assay was based on the separation of ACh and Ch on a polymer gel column followed by passage of the effluent through an IMER, on which the separated ACh and Ch reacted respectively to give each stoichiometric yield of hydrogen peroxide, which was detected electrochemically at a platinum electrode (potential + 0.5 V versus Ag/AgCl). The tip of concentric dialysis probe was made of the semipermeable dialysis membrane of 0.22 mm in outside diameter, and the effective length inserted into rat brain was 3.0 mm. The probe was perfused at a rate of 1 microL/min with Ringer's solution which contained 10 mumol/L (for anesthetized rats) or 1 mumol/L (for freely moving rats) neostigmine, a reversible cholinesterase inhibitor, to elevate ACh level in microdialysate. Before the experiment, the recovery of the probe in vitro was measured at room temperature, and the position of the probe was checked by histological procedure at the end of the experiment. In the range of 0.2-100 mumol/L, the relation between the amounts and the peak areas was linear (r = 0.9988 for ACh and r = 0.9985 for Ch). The detection limit for ACh and Ch, at a S/N ratio of two, was found to be 50 fmol per injection. The probe recoveries(%) for ACh and for Ch were 23.2 +/- 1.4 and 34.3 +/- 3.2(mean +/- SD) respectively. The basal levels of ACh in the microdialysates from striatum and frontal cortex of anesthetized rats as well as from hippocampus and frontal cortex of freely moving rats were 212 +/- 28 and 22 +/- 4 as well as 26 +/- 4 and 83 +/- 7(nmol/L, mean +/- SD, not corrected according to probe recovery) respectively. The perfusion of high concentration K+ (100 mmol/L) through the dialysis probe induced a large increase of ACh in the microdialysates. The critical points for HPLC analysis combined with IMER were briefly discussed. PMID- 11498914 TI - [Enantioseparation of chlorpheniramine and EMD-56431 by micellar electrokinetic capillary chromatography using deoxycholate salt and beta-cyclodextrin]. AB - Two chiral drugs of EMD-56431 and chlorpheniramine are separated by cyclodextrin modified micellar electrokinetic capillary chromatography using beta cyclodextrin(beta-CD) and sodium deoxycholate(SDC) as selector in the paper. All the electrolytes in experiments contain 0.1 mol/L borate. The length of capillary is 45.0 cm as total and 30.0 cm as effective. The running and loading voltages are all 7.8 kV. The effects of pH and concentrations of SDC and beta-CD are studied, in which the best chiral separation conditions for EMD-56431 are pH 10.4, [beta-CD] = 50 mmol/L, [SDC] = 150 mmol/L, and those for chlorpheniramine are pH 9.0, [beta-CD] = 50 mmol/L, [SDC] = 100 mmol/L. The mechanism of chiral separation for the buffer system is initially believed as: the micellar monomer exists almost all as inclusion body with beta-CD, some CD-SDC inclusion complex may exist in micellar because the SDC micellar's gather number is only 4 and the SDC molecular is so big that it can only partly enter beta-CD. Then, the good separation ability of the system is supplied while the ratio of concentration between SDC and beta-CD is in 4:1-4:3; but there will be a optimized total concentration for SDC and beta-CD. The complex interaction among sample, SDC and beta-CD makes intricate change for migration along with the selector's concentration, and the same complex results are also made in pH experiments because of electroosmosis and the acidity of SDC and components. The phenomenon of increasing beta-CD solubility is showed. The beta-CD's solubility with 100 mmol/L SDC can be increased above 150 mmol/L. PMID- 11498915 TI - [Research on the separation behavior of acidic drugs in capillary electrophoresis with reversed direction of electroosmotic flow]. AB - The separation behavior of acidic drugs in capillary electrophoresis with reversal of electroosmotic flow was investigated systematically. Acetylsalicylic acid and its related compound salicylic acid were employed as objective drugs. The cationic surfactant cetyltrimethylammonium bromide (CTAB) was used as reversed reagent for electroosmotic flow. The experimental conditions, such as, cationic surfactant concentration, buffer pH and organic additives, which affected migration time, peak shape and column efficiency, were studied in detail. The experimental results indicated that high speed analysis could be achieved in the capillary electrophoresis with reversed electroosmotic flow induced by cationic surfactant when acidic drugs were analysed. The poor peak shape and low column efficiency caused by the interactions of CTAB with acidic anions would be improved by adding proper organic additive, such as, beta cyclodextrin or acetonitrile. PMID- 11498916 TI - [Development and applications of high performance membrane chromatography]. AB - Development and applications of high performance membrane chromatography (HPMC) are introduced. Base membranes, membrane geometries and separations of biomolecules using high performance affinity membrane chromatography (HPAMC), high performance ion-exchange membrane chromatography (HPIEMC) and high performance hydrophobic interaction membrane chromatography (HPHIMC) are reviewed. Enantiomeric separation using HPMC are prospected. Seventy-six references are cited. PMID- 11498917 TI - [Application of capillary electrophoresis in speciation analysis]. AB - Each species has different chemical and biological behaviors. Toxicity is often dependent on it. So element speciation is important in a variety of environmental, biological, geological, and medical applications. Recently, capillary electrophoresis(CE) has been applied to speciation analysis owing to its high efficiency and short analysis time. This paper reviews the applications of CE-UV and CE-ICP-MS in speciation analysis and discusses some related problems. PMID- 11498918 TI - [Quality control of tobacco flavour by gas chromatography]. AB - In this work, gas chromatography with hydrogen flame detector(FID) and catalytic combustion detector(CCD) was applied to fast analysis of tobacco flavour. The results are satisfactory. Whether the FID or CCD is used, it has its respective characters. Though the sensitivity of GC/CCD is a little bit lower than that of GC/FID, the operation of the former is easier, more convenient and safer. The GC/CCD only uses air as the carrier gas. The column of GC/CCD was SE-54 megabore capillary column(50 m x 0.53 mm i.d.). In this method, all of the interesting flavour compounds are responsed. We have presented some relative correction factors (f1) about flavour compounds in this paper. The satisfactory results show that this new method has certain practical value as an approach of quality control. PMID- 11498919 TI - [Determination of mephenytoin enantiomers in human urine using chiral capillary chromatography and its application in metabolism polymorphism study]. AB - The determination of mephenytoin enantiomers in human urine using chiral capillary chromatography is described. MP in urine was extracted with using dichloroethane and the organic layer was washed with acidic and basic aqueous solution. The organic phase was evaporated to dryness under nitrogen on water bath (55 degrees C). The residue was dissolved in 10 microL ethyl acetate and 1 microL was injected into the GC. MP enantiomers was separated on a Chirasil-Val SFOT column with nitrogen as carrier gas and FID as detector. The linear relationship was obtained over the concentration range of 115-690 micrograms/L (r = 0.9913 for S-MP and r = 0.9934 for R-MP) with a detection limit of 60 ng MP enantiomers/mL urine. The recoveries were 74.41% for S-MP and 73.78% for R-MP. The relative standard deviation within day and between days were less than 6.5%. The method was used to study the metabolism polymorphism of 32 volunteers. The S/R ratio was calculated to express interindividual variation in metabolism, in which, the S/R ratio of 5 subjects was more than 0.95, as the poor metabolizers and that of the others was less than 0.8, as the extensive metabolizers. PMID- 11498920 TI - [Gas chromatographic determination of camphora, mentholum, isoborneol and borneol in Guanxingao]. AB - Guanxingao is a kind of traditional Chinese rubber electuary medicine which is able to either cure or guard against coronary heart disease and angina pectoris. The contents of camphora, mentholum, isoborneol and borneol in Guanxingao are determined by gas chromatography. The purpose of the study is to detect and control the loss of the four volatile components through production and standing and to guarantee the curative effect. Chromatographic analysis was performed on a GC-4004 gas chromatograph(FID). The column was a 3 mm i.d. x 2 m stainless steel tube packed with 7% PEG-1500 on 100-110 mesh 102 non-silanized white support. The column and the FID temperatures were 115 degrees C and 180 degrees C respectively. H2 was the carrier gas, 30 mL/min. Internal standard method was used for the quantitative estimation with naphthalene as the internal standard. The linear ranges were at least within 50-450 mg/L (r = 0.9999, n = 4). The correction factors against naphthalene were 1.262-1.286 and the RSDs were 0.32% 1.5%(n = 12). The recoveries were 98.44%-101.9%. In comparison with the theoretical contents, the average loss percentages are 71.72% (camphora), 65.60% (mentholum) and 66.31% (isoborneol + borneol). The samples were pretreated by means of isothermal (35 degrees C) water-bath extraction with acetone for 6 times with four hours each. PMID- 11498921 TI - [Isolation, purification and determination of the constituents of a toxic protein from plant amenone]. AB - A toxic protein constituent named AME from the stems of plant amenone, have been isolated and purified. Through CM-SFF column chromatography and gel filtration on Sephacryl S-200 column with phosphate saline buffer as mobile phase. All of the operations were performed at 4 degrees C. The pulverized plant amenone material was soaked in phosphate saline buffer, homogenized, left standing overnight and then squeezed through coarse cloth by wringing. The supernatant was applied on the S-SFF column. Then, the column was eluted with the phosphate buffer containing 1 mol/L NaCl. The eluate was collected and dialyzed against water and phosphate buffer. The chromatography of the crude toxin dialyzed was carried out on the CM-SFF column with gradient elution of phosphate buffer containing NaCl. The fourth peak was collected and then applied on a gel filtration Sephacryl S 200 column using neutral phosphate buffer as mobile phase. The protein was further separated on two connected Protein 125 columns with mobile phase of 0.2 mol/L phosphate buffer (pH 6.5), the eluate was monitored at 280 nm on photodiode array detector. The protein presents typical the absorption spectrum of protein in ultraviolet region with the strong absorption at 280 nm and the weak absorption at 260 nm. The purity of peak of the protein was judged from the spectrum. The molecular weight of AME measured by two connected protein columns was approximately 35,000 D. The composition of amino acids was determined with OPA post-column derivatization/fluorescence detection. PMID- 11498922 TI - [Separation and determination of eight plant hormones by reversed-phase high performance liquid chromatography]. AB - In this paper, reversed-phase high performance liquid chromatographic technique was used for the separation and determination of eight plant hormones. Methanol water-acetic acid system was chosen as the mobile phase. The effects of different separation conditions, such as the methanol and acetic acid concentrations in mobile phase, on the retention behaviours of eight plant hormones in this system were studied. The general trends in retention behaviours could be correlated to the methanol concentration in mobile phase. The experimental results showed that the optimum separation was achieved with following gradient elution condition: 0 3 minutes, 70% (water percentage in mobile phase), 3-13 minutes, 70%-20%, 13-48 minutes, 20%. Benzene was added to be as the internal standard. Under this experimental condition, the eight plant hormones could be separated completely and detected quantitatively at 260 nm within 16 minutes. The calibration curves for the eight compounds gave linearity over a wide range. The correlation coefficients of each components were r(ZT) = 0.9971, r(GAs) = 0.9999, r(K) = 0.9997, r(BA) = 0.9995, r(IAA) = 0.9998, r(IPA) = 0.9982, r(IBA) = 0.9995 and r(NAA) = 0.9995. The method is rapid, simple and efficient. It is a suitable method for the accurate determination of gibberellic acid (GA) and alpha naphthaleneacetic acid (alpha-NAA) in products for agricultural use. PMID- 11498923 TI - [Separation of celiprolol enantiomer by high performance liquid chromatography with urea derivative as chiral stationary phase]. AB - Celiprolol enantiomer was resolved directly by using normal-phase HPLC with urea derivative as chiral stationary phase (CSP). The resolution condition was optimized by varying the content of ethanol and 1, 2-dichloroethane in the mobile phase. The effects of the two components on stereoselectivity factor (alpha) and stereochemical resolution factor (Rs) are demonstrated. The higher the 1,2 dichloroethane content the faster the elution of solute is, but the lower the values of alpha and Rs are. For a suitable content of ethanol in mobile phase, the maximum resolution factor (Rs) can be obtained. Ethanol is a strong proton donor and proton-acceptor. Its strong hydrogen bond interaction with solute and CSP is important for the direct resolution. In order to obtain both the low retention time and a high Rs, we chose the mobile phase with n-hexane:1,2 dichloroethane:ethanol (V/V/V) = 77:21:2. Other organic modifiers such as methanol, iso-propanol, n-butanol and acetonitrile were also used. Iso-propanol, methanol and n-butanol showed longer retention time and lower values of alpha and Rs than ethanol. Acetonitrile is only a proton-acceptor and has weak hydrogen bond interaction with solute and CSP, so resolution wasn't obtained. The elution order of enantiomer was also discussed. We thought that the hydrogen bond interaction between solute and the (S)-val component of CSP may mainly control the chiral recognition. The interaction strength is different between (R)- and (S)-celiprolol, so the celiprolol enantiomer was resolved. And the elution order is S, R. PMID- 11498924 TI - [Enantiomeric high performance liquid chromatographic separation of epinephrine and analogous using chiral mobile phase additives]. AB - The enantiomeric resolution of epinephrine, isoproterenol, ephedrine was studied by reversed phase HPLC. beta-CD, DM-beta-CD, TM-beta-CD were used as chiral mobile phase additives. The effects of the concentration of DM-beta-CD and the concentration of methanol on resolution of D,L-epinephrine were investigated. The results showed: D,L-epinephrine and D,L-isoproterenol can be separated by beta CD, DM-beta-CD, TM-beta-CD, but D,L-ephedrine can't. Hydrogen bonding plays an important role in the chiral recognition mechanism. In the separation of isoproterenol, the enantioselectivity of TM-beta-CD was better than those of beta CD and DM-beta-CD. As the concentration of DM-beta-CD changed from 0.04 mmol/L to 1.00 mmol/L, the resolution varied slightly. The experiment of the effect of methanol concentration on Rs showed: when the ratio of methanol/water was 40/60, the Rs reached the maximum. The coefficients of variation of retention time of D epinephrine and L-epinephrine were 1.3% and 1.4% respectively. It is demonstrated that the chromatographic systems with a dynamically-generated stationary phase with methylated-beta-cyclodextrin are versatile tools for enantiomer separation. This technique leads to column with excellent time stability and good reproducibility of the enantio-selectivity. PMID- 11498925 TI - [Simultaneous determinations of carbamate pesticides in cereal by high performance liquid chromatography with post-column fluorescence derivatization]. AB - A liquid chromatographic method for the simultaneous determination of carbamate pesticides in cereal by Waters Carbamate Analysis System with post-column fluorescence derivatization is described. The cereal sample homogenate was extracted with acetone and water. The extract was partitioned between 5% sodium chloride solution and dichloromethane. The dichloromethane layer was concentrated to dryness. The residue was dissolved in n-hexane and extracted with acetonitrile (saturated with n-hexane). The acetonitrile layer was decreased with n-hexane (saturated with acetonitrile), then the acetonitrile layer was concentrated to dryness. The residue was dissolved in methanol. It's the test solution for HPLC. Nine carbamate pesticides and their three metabolites were separated by Waters Carbamate Column with gradient elution. Two steps underwent in the post-column system: hydrolysis by NaOH solution and the reaction of the product with o phthalaldehyde and 2-mercaptoethanol. The resulting fluorescence derivative was detected at 445 nm (excitation at 339 nm). The selectivity, reproducibility and sensitivity of proposed method are better than those reported with other methods. The detection limits was 5 micrograms/kg of samples. PMID- 11498926 TI - [Determination of sisomicin in body fluid by high performance liquid chromatography-indirect photometric detection method]. AB - A high performance liquid chromatography-indirect photometric detection (HPLC IPD) method for determination of sisomicin sulfate in body fluid (serum, urine, saliva, cerebrospinal fluid and fluid oozed out from wound) was established. The conditions in this method were a mobile phase solution of methyl alcohol acetonitrile-water (20:10:70) containing nicotinamide 0.5 mmol/L, sodium 1 heptanesulfonate 5 mmol/L and phosphoric acid 0.05 mol/L, a Spherisorb C18 column (4.6 mm x 250 mm) and an UV detector with sensitivity of 0.05 Aufs (0.1 Aufs for urine). The detection wavelength was fixed at 268 nm. The average recovery for sisomicin in serum was 96.92% +/- 4.63% and the coefficients of variation were 4.75% and 5.65% for within-day and day-to-day tests respectively. The detectable limit was 0.1 ng. The concentration of sisomicin in body fluid of 4 patients was determined. PMID- 11498927 TI - [The determination of melatonin in health-caring medicine for specific purposes by high performance liquid chromatography-mass spectrometry]. AB - A rapid and precise method for the determination of melatonin in health-caring medicine for specific purposes, based on high performance liquid chromatography mass spectrometry, has been developed. The compound was identified using DAD UV spectrum and mass spectrum. Hypersil ODS column, 100 mm x 2.1 mm i.d., and mobile phase of methanol-water (70:30, V/V) were used for the assays. The quantitation was performed with external standard, and detection at 275 nm. The calibration curve of melatonin was linear in the range of 5-50 mg/L and the detection limit was ca. 0.2 ng. PMID- 11498928 TI - [The rapid effects of steroids on glycine uptake in neuroblastoma cell strain SK N-SH cells]. AB - In the present study, glycine uptake in SK-N-SH cells was determined with liquid scintillation technique, and the rapid effects of steroids on glycine uptake in SK-N-SH cells were investigated. The results were as follows. High-affinity glycine uptake in SK-N-SH cells was dependent on Na+ and Cl-. Corticosterone (CORT), progesterone (P) and dexamethasone (DEX) had rapid effects on the glycine uptake. Since estradiol (E2) and deoxycorticosterone (DOC) had no effects, it was suggested that the rapid effects of steroids were specific. The rapid effects of CORT were concentration-dependent in a range of 10(-9)-10(-6) mol/L. The rapid effects were not affected by the inhibitor of protein synthesis and persisted even when CORT was conjugated with bovine serum album, but attenuated when Ca2+ was absent in the external medium. The results suggest that the steroid effect on glycine uptake in SK-N-SH cells was nongenomicly mediated. PMID- 11498929 TI - [Effect of scorpion venom on the release of GABA in hippocampus of epileptic rats induced by kainic acid]. AB - In the present study, the effect of scorpion venom (SV) on epileptic susceptibility and its neurochemical mechanisms were investigated. After SV or normal saline (NS) administration for three weeks in epileptic rats, epileptic seizures were much more decreased in kainic acid (KA) + SV group than those of KA + NS group. GABA immunocytochemistry showed that both (gamma-aminobutyric acid immunoreactivity (GABA-IR) density and the number of GABA positive neurons were markedly decreased in the ventral hippocampus, especially in hilus of KA + NS group as compared with NS + NS group. However, in the group treated with SV for three weeks, the number of GABA positive neurons and the density of GABA-IR were greater increased than those of both KA + NS group and the NS + NS group. The number of GABA positive neurons and the density of GABA-IR were unchanged in the dorsal hippocampus of all the four groups. The above results suggest that SV may selectively prevent loss of GABAergic interneurons in the ventral hippocampus of rats susceptible to epileptic seizures and increase the release of GABA as an antiepileptic mechanism. PMID- 11498930 TI - [Protein kinase a mediated excitatory adrenergic effect on chronically compressed dorsal root ganglion neurons in rats]. AB - With a model of chronically compressed dorsal root ganglion (CCD), the present study was undertaken to test how the plasticity of sympathetic-sensory coupling is and whether the coupling is mediated by intracellular messenger PKA by analysing extracellularly recorded spontaneous activity of single A-fibers originating from the CCD neurons in vitro. Eighty-five out of 95 neurons from injured DRGs during application of norepinephrine (NE) were adrenosensitive. Among the 85 neurons, 44 exhibited excitation, 21 showing excitation followed by suppression, 6 displaying alternated excitation and suppression, and 14 suppression. In addition, adrenosensitivity was observed in 15 silent injured DRGs. The excitatory effect of NE was blocked by alpha 1 and alpha 2 adrenoceptor antagonists yohimbine (10 mumol/L) or prazosin (5 mumol/L). Rp-cAMPS (50-250 mumol/L, n = 6), a specific inhibitor of PKA, and H-89 (10 mumol/L, n = 6), an inhibitor of PKA catalytic subunit, obviously suppressed the NE-evoked excitation. Furthermore, the excitatory effect of NE was attenuated by SQ 22, 536 (1 mmol/L), an adenylate cyclase inhibitor (n = 6). The above results demonstrate that injury to DRG neuron body triggered the adrenosensitivity, which was mediated by alpha 1, alpha 2 adrenoceptors and PKA. PMID- 11498931 TI - [Effects of L-NNA on postsynaptically evoked potential of oculomotoneurons in cats]. AB - The present study was performed to explore the effect of endogenous nitric oxide (NO) on synaptic transmission between neurons located in Forel's field H (FFH) and vertical oculomotoneurons (OMNs) innervating the inferior oblique (IO) muscle in cats. The amplitude of monosynaptic excitatory field potential in the IO OMNs pool induced by stimulating FFH began to decrease gradually 2-3 min after intracarotid administration of L-NNA, an NO synthase inhibitor, reached its minimum by 6-10 min, and was followed by a slow recovery. This effect of L-NNA could be prevented by pre-administration of NO precursor L-arginine, without changing the field potential by itself. The results suggest that there is an endogenous NO release in the oculomotor nuclear, which can potentiate synaptic transmission between FFH neurons and OMNs. PMID- 11498932 TI - [Changes in blood pressure in normal rats transplanted with kidney of SHR]. AB - The experiments were performed on spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats of 12 weeks old. Systolic blood pressure in WKY recipients receiving an SHR or WKY rat kidney was observed and the influence of immuoinhibitor cyclosporin (CsA) on blood pressure was investigated. The systolic blood pressure (SBP) of tail artery was measured by tail cuff technique in 6 WKY recipients of one SHR kidney and 6 WKY recipients of a WKY kidney from before transplantation to 5 weeks after transplantation. At the time when kidneys were harvested for transplantation from 12-week-old SHR and WKY rats, the SBP in WKY recipients was not significantly different. The SBP in WKY recipients of an SHR kidney was significantly higher than that in WKY recipients of a WKY kidney at 3, 4 and 5 weeks after transplantation. The immunoinhibitor CsA exerted no effect on the arterial blood pressure in either intact or uninephrectomized SHR and WKY rats. The results indicate that the kidney of SHR plays an important role in the pathogenesis of genetic hypertension. PMID- 11498933 TI - [Effect of recombinant human interleukin-1 beta on K+ channels of mice bone marrow stromal cells]. AB - In the present study, both cell-attached and inside-out mode of patch clamp technique were applied to detect the effect of recombinant human interleukin-1 beta (IL-1 beta) on K+ channels of mice bone marrow stromal cells. A 16.7 pS voltage-dependent K+ channel resembling the delayed rectifier K+ channel in excitable cells was identified and characterized. Under stimulation of IL-1 beta (1000 U/ml), the single channel conductance increased to 26.1 +/- 3.6 pS (P < 0.01). IL-1 beta also markedly increased the open time constant and the open probability, but decreased the closed time constant as compared to the same patches without IL-1 beta. In addition, IL-1 beta induced multi-channel activities in about 80 percent patches by inducing more K+ channels of the same type open. These results suggest that activation of K+ channels plays a role in signal transduction of interleukin-1. PMID- 11498934 TI - [Effects of bone marrow endothelial cell conditioned medium and its ultrafiltration-prepared components on the growth of CFU-GM]. AB - In this study, we obtained the serum-free conditioned medium from subcultures of murine bone-marrow-derived endothelial cell line which has been established recently by ourselves. And then, molecular weight (MW) > 10 kD, 3-10 kD, 1-3 kD, 0.5-1 kD and < 0.05 kD components were sifted out from the conditioned medium (mBMEC-CM) by means of serial ultrafiltration. Assays of granulocyte-macrophage colony-forming cell (CFU-GM) were performed to examine the effects of mBMEC-CM and its ultrafiltration-prepared components. It was not observed that mBMEC-CM and MW 3-10 kD component had any significant influence on the growth of CFU-GM. However, MW > 10 kD and 0.5-1 kD components enhanced the proliferation of CFU-GM, whereas MW 1-3 kD and < 0.5 kD ones inhibited it. All these four components exerted their effects on the growth of CFU-GM in dose-dependent way. Our observations suggest that under the conditions of in vitro, murine bone marrow endothelial cells produce several active components which have promoting or inhibitory effects on the growth of CFU-GM. PMID- 11498935 TI - Biphasic activation of renal afferent by intrarenal artery injection of bradykinin in anesthetized rabbits. AB - The effect of intrarenal artery injection of bradykinin (BK, 5.0 micrograms/kg) on multi- and single-unit recordings of afferent renal nerve activity (ARNA) was examined in anesthetized 48 rabbits. The results obtained are as follows. (1) There were two phases of increase in ARNA following intrarenal BK. The early phase occurred immediately while the delayed phase made its appearance about 7 min later. The degree of increase in ARNA of the delayed phase induced by intrarenal BK was more prominent than that in the early phase. BP was actually unaltered following intrarenal BK. (2) By pretreatment with indomethacin (Indo, 5.0 mg/kg), the delayed phase of increase in ARNA induced by intrarenal BK was attenuated, while the early phase was not affected. (3) Pretreatment with L-NAME (30 mg/kg) led the delayed phase to be blocked completely while the early phase was partially decreased. From the above-mentioned observations, it is concluded that intrarenal BK induces a significant increase in ARNA in two phases. The early phase may be due to the direct action of BK and partially due to the NO action, while the delayed phase may be attributed to the action of released prostaglandin and NO as a result of intrarenal BK. PMID- 11498936 TI - [The role of nitric oxide in the angiotensin II-induced hypertrophy of cardiac myocytes]. AB - The purpose of this study was to determine the role of nitric oxide (NO) in the angiotensin II-induced hypertrophic response in cultured neonatal rat cardiac myocytes. Angiotensin II induced significant increase of protein content, decrease of NOS activity in cultured neonatal rat cardiomyocytes and decrease of NO concentration in the culture medium. Angiotensin II decreased significantly the eNOS mRNA level of cardiomyocytes. Saralasin and PTX inhibited significantly the increased protein content, the decreased NOS activity and the decreaseed NO production of cardiomyocytes induced by angiotensin II. Sodium nitroprusside increased significantly NO concentration in the culture medium and inhibited significantly the increased protein content of cardiomyocytes induced by angiotensin II. The results suggest that angiotensin II may induce hypertrophy of cultured neonatal rat cardiomyocytes. The effect of angiotensin II is mediated by receptors which are coupled with PTX-sensitive G protein and may be related to the decreased eNOS gene expression, NOS activity and NO production. Exogenous NO can prevent the hypertrophy of cardiomyocytes induced by angiotensin II. PMID- 11498937 TI - Effect of intracarotid administration of adenosine on the activity of area postrema neurons in barodenervated rats. AB - To observe the effect of intracarotid administration of adenosine on the electrical activity of area postrema (AP) neurons, 76 spontaneous active units were recorded from 45 sino-aortic denervated Sprague-Dawley rats using extracellular recording technique. The results obtained are as follows. (1) Following intracarotid administration of adenosine (Ado, 25 micrograms/kg), the discharge rate of 29 out of 42 units decreased markedly from 6.26 +/- 0.75 to 4.74 +/- 0.76 spikes/s (P < 0.01), whereas that of 6 units increased from 4.13 +/ 0.77 to 4.72 +/- 0.83 spikes/s (P < 0.05), and the other 7 showed no response. Blood pressure (BP) and heart rate (HR) were unaltered throughout the experiment. (2) 8-phenyltheophylline (8-PT, 15 micrograms/kg), a nonselective adenosine receptor antagonist, completely blocked the inhibitory effect of Ado in 10 units. (3) Selective A1 adenosine receptor antagonist, 8-cyclopentyl-1,3 dipropylxanthine (DPCPX, 50 micrograms/kg), blocked the effect of Ado in 12 units to a remarkable extent. (4) Glibenclamide (500 micrograms/kg), a blocker of ATP sensitive potassium channel, abolished the effect of Ado in 12 units. The above results indicate that Ado can inhibit spontaneous electrical activity of AP neurons, which is mediated by adenosine A1-receptor with the involvement of ATP sensitive potassium channels. PMID- 11498939 TI - [Changes of inhibitory amino acid in the monkey frontal cortex during a figure recognition guided sequential movement task]. AB - The levels of gamma-aminobutyric acid (GABA) and glycine (Gly) in the micro dialysate from the monkey frontal cortex were measured by using the techniques of intracerebral microdialysis and high performance liquid chromatography with fluorescent detection while the animal was performing the Figure Recognition Motor Sequence (FRS) task. The probes were sited in the prefrontal cortex (PFC, 46 area), premotor cortex (PM, F7 and F2 areas) and primary motor cortex (MI, F1 area). The results showed that the level of GABA in PFC increased during the FRS task, whereas the level of Gly showed no significant changes. PMID- 11498938 TI - [Expression of prolactin, TGF alpha and TGF beta 1 genes in estrogen-induced eutopic and ectopic pituitary prolactin-secreting tumors of rats]. AB - The Sprague-Dawley (SD) rat bearing a heteroplasted pituitary underneath renal capsule was used to observe differential expression of prolactin (PRL), transforming growth factor alpha (TGF alpha) and transforming growth factor beta 1 (TGF beta 1) genes during the formation of pituitary prolactin-secreting tumor (prolactinoma) induced by 17 beta-estradiol (E2). Our results indicated that in both eutopic and ectopic pituitaries disconnected from hypothalamus formed simultaneously PRL-secreting tumors after the rats treated with E2 for 120 days in vivo, which was accompanied by overexpression of PRL gene (P < 0.05-0.01). The PRL mRNA level was higher in eutopic prolactinoma than that in ectopic prolactinoma (P < 0.05). Overexpression of TGF alpha and TGF beta 1 genes were also detected in eutopic prolactinoma. However, the expression of TGF alpha and TGF beta 1 genes in ectopic prolactinoma was similar to that in normal pituitary. It is suggested that TGF alpha and TGF beta 1 may be involved in prolactinoma tumorigenesis of eutopic pituitary. However, the mechanism mediating eutopic and ectopic prolactin-secreting tumor formation seems different. PMID- 11498940 TI - [Effects of intracarotid injection of capsaicin on electrical activity of rostral ventrolateral medullary neurons in rats]. AB - The effects of intracarotid injection of capsaicin on spontaneous electrical activity of nucleus paragigantocellularis lateralis (PGL) neurons in rostral ventrolateral medulla (RVLM) were examined in 35 anesthetized rats with sino aortic denervation by using extracellular recording technique. The results obtained were as follows: (1) In response to intracarotid injection of capsaicin (10 mumol, 0.1 ml), MAP was increased from 10.74 +/- 0.13 to 12.56 +/- 0.21 kPa (P < 0.001), and HR from 374 +/- 4 to 395 +/- 5 bpm (P < 0.001). All of 30 PGL neurons recorded responded to intracarotid injection of capsaicin with an increase in spontaneous discharge rate from 12.6 +/- 0.7 to 20.9 +/- 1.1 spikes/s (P < 0.001); (2) In 10 units, the excitatory response of PGL neurons to intracarotid injection of capsaicin was significantly inhibited by pretreatment with the vanilloid receptor (capsaicin receptor) antagonist ruthenium red (200 mmol, 0.1 ml). These results suggest that capsaicin may stimulate the vanilloid receptor of neurons in RVLM, thereby resulting in activation of PGL neurons. PMID- 11498941 TI - [A method for preparation of carbon fiber electrode]. AB - Carbon fiber electrode (CFE) can be used in detection of exocytosis of single neuron or endocrine cell. A simplified method for preparation of CFE, as described in the present paper, greatly ensures the coordination and success of the preparation. Such CFEs have a low noise level. Exocytosis of rat adrenal chromaffin cells was investigated in the clarification of the effect of MPP+ on transmitter release. It is found that exocytosis was not stimulated by MPP+ and the high K+ induced secretion was not changed by MPP+. These observations suggest that the previously reported elevation of dopamine content in the surrounding histosolution of DA neuron is probably resulted from blockage of the re-uptake of DA by MPP+. PMID- 11498942 TI - [Simultaneous observation of parameter of intraventricular pressure and energy metabolism in rat hearts using nuclear magnetic resonance]. AB - A technique for simultaneous observation of parameters of intraventricular pressure and energy metabolic status in isovolumic rat hearts was described. The parameters of intraventricular pressure were collected and processed using a computerized monitoring system for heart function, and myocardial energy metabolism was investigated by collecting the 31P NMR spectra. The preparation was stable metabolically and functionally for 2 h under the NMR experimental conditions. PMID- 11498943 TI - [Pattern and dynamic changes of integer multiples in spontaneous discharge of injured dorsal root ganglion neurons]. AB - For the purpose of the present investigation, spontaneous discharges of dorsal root ganglion (DRG) neurons of the rats, which had undergone 3-10 days' chronic compression, were studied. It was found that the interspike interval (ISI) of 17 out of 156 fibers had integer multiples temporal rhythm pattern, in which all the ISI were integer multiples of a basic ISI and a return map of their ISI could be constructed as a crystal grid structure. This temporal pattern could be modified by Na+ channel and K+ channel on their membranes. These data indicated the presence of some irregular discharge trains with their intrinsic regularity. PMID- 11498944 TI - [Changes in the mRNA concentration of preprocholecystokinin in rat cerebral cortex and hippocampus after gastric distension]. AB - Cholecyctokinin-octapeptide (CCK-8) is one of the brain-gut peptides widely distributed in neurons of the central nervous system, especially in cerebral cortex and hippocampus. This peptide is thought as a neurotransmitter or modulator. The role of central CCK in the integration of gastric motility and food intake is still unclear. The present investigation was undertaken to see whether the gastric distension exerts some effect on the expression of CCK mRNA in cerebral cortex and hippocampus by means of in situ hybridization and semi quantitative RT-PCR techniques. The results are as follows. (1) Discrete positive hybridization cells were seen in cerebral cortex and hippocampus in control group. (2) Significant increases of CCK mRNA expression in cerebral cortex and hippocampus produced by gastric distention observed by increased of the volume, the number, the IOD and the SA of positive cells. (3) Amplified bands of the expected size of 158 base pairs were found in cerebral cortex and hippocampus, and signals of gastric distention group were higher than those of control group. The results from RT-PCR assay are in agreement with those based on in situ hybridization. These results imply that biosynthesis of CCK-containing neurons in cerebral cortex and hippocampus is accelerated by afferent signals from gastrointestinal system and may be involved in regulation of gastric motility and food intake. PMID- 11498945 TI - [Effect of dietary zinc on microtubule-associated protein 2 expression in the brain of mice]. AB - Zinc deficiency could result in reduction of microtubule polymerization, which may cause impairment of brain development and function. The relationship between zinc deficiency and microtubules polymerization is still unclear. In this paper, microtubule-associated protein 2 (MAP2) expression in the brain was examined in order to explore the mechanism of zinc regulated microtubule polymerization. 80 pregnant ICR mice, randomized into 5 groups, were fed with experimental diets of different zinc levels (from 1 to 100 mg/kg) during pregnancy and lactation. The MAP2 expression in the brain of offsprings was examined by Western blot assays. The results showed that MAP2, including MAP2a, MAP2b and MAP2c, were expressed in brain from embryonic day 15, but not found on embryonic day 10. The high molecular weight of MAP2a and MAP2b expressed continuously from embryonic day 15 to postnatal day 70 (adult). While the low molecular weight of MAP2c was down regulated from embryonic day 15 to non-existing on postnatal day 70. The expression of MAP2 in cerebrum and cerebellum kept closely at the positive dependence with dietary zinc level. The order of the levels of expression of MAP2 of the various groups administrated with different amounts of zinc is as follows: 1 mg/kg < 5 mg/kg < 30 mg/kg < 100 mg/kg. The above results suggest that zinc deficiency may inhibit the MAP2 expression, while zinc supplement exerts much improvement. The lowered level of MAP2 expression is one of important mechanisms underlying impairments of microtubule polymerization, as a result of zinc deficiency. PMID- 11498946 TI - [Ciliary neurotrophic factor prevents NO mediated cytotoxicity in cultured hippocampal neurons]. AB - The effects of ciliary neurotrophic factor on nitric oxide (NO)-induced cytotoxicity were examined in primary cultures of rat hippocampal neurons. The neurotoxicity was induced by NO donors, such as sodium nitroprusside (SNP), s nitroso-N-acetylpenicillamine (SNAP), NOS substrate, L-arginine (L-Arg) and ionomycin (a calcium ionophore). Preincubation of the cultures with ciliary neurotrophic factor (CNTF) for 24 h promoted neuronal survival and decreased the LDH efflux and the level of NO induced by ionomycin. The protective effects of CNTF against NO cytotoxicity were concentration-dependent. The above findings suggest that CNTF promotes the survival of neurons in the hippocampus by preventing NO mediated neurotoxicity. PMID- 11498947 TI - [Effect of electroacupuncture and 7-NI on penicillin-induced epilepsy and their relation with intrahippocampal NO changes]. AB - In order to investigate the effect of 7-nitroindazole (7-NI) on penicillin induced epilepsy and the relationship between NO and anti-epileptic effect of electro-acupuncture, computerized physiological polygraph and NO-sensitive electrode with potentiostat were respectively used to record the total power spectrum (TPS) of EEG and the concentration of NO in rat hippocampus pretreated with electro-acupuncture or 7-NI for 30 min before penicillin. The results showed that the TPS of EEG and concentration of NO in hippocampus increased greatly after the epilepsy induction. Treatment with electroacupuncture inhibited the TPS of EEG sharply (P < 0.01). The latency and threshold of epilepsy induction were decreased by 7-NI, but paroxysm abated and the TPS was inhibited (P < 0.05) as compared with the control group. Both pretreatments with 7-NI and electroacupuncture decreased the NO concentration (P < 0.01). The above results suggest that the propagation of penicillin-induced seizures is facilitated by nNOS, but with some increase in the latency and threshold of induction. Electroacupunture may inhibit seizure through decreasing nNOS transcription in hippocampus. PMID- 11498948 TI - [Comparison of vasorelaxing actions of vasonatrin peptide, C-type natriuretic peptide and atrial natriuretic peptide]. AB - The vasorelaxing effects of vasonatrin peptide (VNP), C-type natriuretic peptide (CNP) and atrial natriuretic peptide (ANP) on isolated rat pulmonary artery, abdominal aorta and celiac vein were measured by in vitro perfusion. The results showed that VNP, CNP and ANP caused concentration-dependent relaxation in isolated rat pulmonary artery, abdominal aorta and celiac vein with endothelium or without endothelium. The maximal responses (Rmax) of VNP were (76 +/- 17)%, (51 +/- 14)% and (62 +/- 14)% in pulmonary artery, abdominal aorta and celiac vein with endothelium respectively, whereas those of CNP were (31 +/- 8)%, (22 +/ 7)% and (41 +/- 8)%, and ANP (38 +/- 10)%, (41 +/- 10)% and (11 +/- 4)%. The median effective concentration (EC50) of VNP were 16 +/- 11, 35 +/- 18 and 12 +/- 8 nmol/L in pulmonary artery, abdominal aorta and celiac vein with endothelium respectively, while those of CNP were 148 +/- 112, 299 +/- 84 and 14 +/- 12 nmol/L, and ANP 66 +/- 47, 16 +/- 15 and 909 +/- 445 nmol/L. VNP were more effective than CNP and ANP, and the differences were statistically significant (P < 0.05-0.01). The potency of these peptides for relaxing the blood vessels can be summarized as: VNP > ANP > or = CNP for pulmonary artery; VNP > ANP > CNP for abdominal aorta; VNP > CNP > ANP for celiac vein. There was no significant difference between vessels with intact endothelium and those denuded of endothelium (P > 0.05). PMID- 11498949 TI - [Effect of angiotensin II on c-fos expression and protein synthesis in cultured rat myocardial cells]. AB - The present study was to investigate the effects of angiotensin II on c-fos mRNA expression and protein synthesis in cultured neonatal rat myocardial cells. The results showed that angiotensin II induced c-fos mRNA expression, increased protein content in a dose-dependent manner and stimulated 3H-leucine incorporation rate. All these effects were blocked by angiotensin II receptor antagonist saralasin. The angiotensin II-induced expression of c-fos gene was also blocked by Ca2+ channel antagonist nicardipine. PMID- 11498950 TI - [Effect of chronic hypoxia on the calcium and potassium current of myocytes of the right ventricle in guinea pig]. AB - To explore the effect of chronic hypoxia on calcium and potassium currents of the right ventricle myocytes in the guinea pig, the cell membrane capacities, calcium and delayed rectifier potassium current were recorded using the whole-cell patch clamp technique from single right ventricle myocytes in guinea pigs under chronic hypoxia. The result demonstrated that the cell membrane capacities of the hypoxic group were larger than those of the control; between test potential of -20 mV to +20 mV, the calcium current density of the myocytes of the chronic hypoxic group was less than that of the control; the amplitude of IK of the myocytes of the chronic hypoxia group were less than that of the control group between test potential of +20 mV to +60 mV, and the IK density of the hypoxic group was also less than that of the control group between -20 mV to +60 mV. Consequently, it is suggested that chronic hypoxia may cause an increase of the membrane capacity, a decrease of the calcium current density of ventricle myocytes and a delay of the rectifier potassium current amplitude and density. All these observations provide information for the ionic basis of the attenuated contraction of cardiac muscles and prolonged action potential duration of hypoxic guinea pigs. PMID- 11498951 TI - [Preventive effect of endothelin-1 mRNA antisense oligonucleotide on acute myocardial ischemic arrhythmia in rats]. AB - Acute myocardial ischemia was induced by occlusion of the anterior descending of left coronary artery (LAD) in rats; the resultant arrhythmia in 1 h after LAD occlusion was evaluated. In order to prevent expression of endothelin-1 mRNA, human endothelin-1 mRNA antisense oligonucleotide (ET-1 AS-ODN) was intravenously injected 2 h before LAD occlusion. The effect of AS-ODN on plasma ET-1 concentration and the acute ischemic arrhythmia were observed. The results showed that plasma ET-1 was significantly decreased in rats pretreated with AS-ODN, and both the incidence and severity of the acute ischemic arrhythmia were decreased dose-dependently as compared with normal saline control and sense oligonucleotide control, indicating that ET-1 AS-ODN could prevent acute myocardial ischemic arrhythmia and that endogenous endothelin-1 may play an important role in the development of acute ischemic arrhythmia in rats. PMID- 11498952 TI - [Changes of osteopontin and matrix Gla protein mRNA expressions in the healing process of rat thoracic aorta damaged by balloon angioplasty]. AB - Using Northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR) the dynamic changes of osteopontin (OPN) and matrix Gla protein (MGP) mRNA expression in the healing process of rat thoracic aorta damaged by balloon angioplasty were investigated. The results showed that expression of OPN and MGP in the thoracic aorta damaged group was higher compared with normal group. 1, 7, and 14 d after thoracic aorta was damaged, expression of OPN and MGP was increased gradually, and decreased after artery damaged 21 d. PMID- 11498953 TI - [Changes in the content and activity of HSC70/HSP68, proteinases and phosphatases during liver regeneration]. AB - This article reports changes in acid and alkaline phosphatases (ACP and AKP), constitutive heat shock protein 70/induced heat shock protein 68 (HSC70/HSP68) and acid and neutral proteinases during liver regeneration (0-144 h) after 2/3 hepatectomy (partial hepatectomy, PH). Both ACP and AKP had two active peaks at 4 and 48 h, 16 and 96 h, respectively, which were followed by significant decrease. The content of HSC70/HSP68 also showed two peaks (16 and 96 h), of which the content after the second peak decreased more obviously than after the first peak. Moreover a 90 kD neutral proteinase was induced at the time from 2 to 6 h and a 27 kD acid proteinase was induced at 36 h. The results suggest that the ACP and the 90 kD neutral proteinase may participate in activating hepatocytes from G0 phase into G1-phase, and AKP and HSC70/HSP68 may play a role mainly in DNA synthesis, cellular metabolism and proliferation. Furthermore 27 kD acid proteinase may be involved in re-differentiation of hepatocytes and reconstruction of liver tissue. PMID- 11498954 TI - [Inhibition of gastric myoelectric activity and gastric motility by microinjection of substance P into dorsal vagal nucleus in rats]. AB - The present investigation showed that microinjection of substance P (SP) into dorsal vagal nucleus inhibited gastric myoelectric fast wave and gastric motility. The effect could be blocked by SP antiserum or SP antagonist [Arg6, D Trp7,9, N-Me-Phe8]-SP6-11 or vagotomy. Depletion of sympathetic transmitters by reserpine did not affect induction of the inhibition. The results indicate that both exogenous and endogenous SP of dorsal vagal nucleus decrease the gastric myoelectric fast wave and motility, which is mediated by vagus nerve. PMID- 11498955 TI - [Role of M-type receptor in internal calcium release and quantal secretion in rat adrenal chromaffin cells]. AB - In rat single adrenal chromaffin cells, the effects of methacholine (MCh) on [Ca2+]i and catecholamine secretion were studied with fura-2 fluorescence and carbon fiber electrodes. In the presence of 2 mmol/L Ca2+ in the bath, locally applying 1 mmol/L MCh, either containing or not containing Ca2+, evoked both [Ca2+]i and secretion signals. In the absence of Ca2+ in the bath, MCh could still evoke [Ca2+]i and secretion. These results suggest that MCh causes release of Ca2+ from Ca2+ stores, which is sufficient to yield the evoked secretion. The Ca2+ store can be depleted by single MCh puff in the absence of Ca2+ in the bath. PMID- 11498956 TI - A patch clamp study on reconstituted calcium permeable channels of human sperm plasma membranes. AB - Ionic flux is thought to be important in the initiating process of gamete interaction such as acrosome reaction. However, modern electrophysiological methods, intracellular recording and patch-clamping, are difficult to approach the ion channels in mammal sperm membrane of an intact sperm due to its small size. In this work, by reconstituting the channel protein into lipid bilayer, Ca2+ channels in human spermatozoa were investigated with voltage clamp technique. Membrane proteins isolated from human sperm of 12 healthy donors were incorporated into lipid bilayer via fusion. In a cis 50//trans 10 mmol/L CaCl2 solution system, two types of channel events with similar reversal potential near the value of a perfect Ca2+ electrode, and sensitive to nifedipine and verapamil, were observed. Their unit conductance was 40 and 25 pS respectively. Percentage of channel open time was not dependent to holding potential for the former. However, for the channels of 25 pS, the percentage increased when the holding potential was changed from -20 to 100 mV. Ca(2+)-permeable channels were also detected from the spermatozoon samples of two infertile donors. Abnormal open time of these channels indicates that there are some defects in the conformation of the channel protein of infertile sperm membrane. PMID- 11498957 TI - [Effect of OFQ injection into intracerebroventricular and preoptic area on blood pressure and heart rate in rats]. AB - Orphanin FQ (OFQ) is a novel peptide comprised of an amino acid sequence very similar to that of dynorphin A. In the present investigation the effect of OFQ on cardiovascular activities was studied. Introcerebroventricular (icv) injection of OFQ at doses of 1 and 10 micrograms produced significant decrease in heart rate (HR) and mean arterial pressure (MAP). Icv pretreatment with naloxone could not prevent the hypotensive and bradycardial response produced by 1 microgram OFQ. Injection of 1 microgram OFQ preoptic area (POA) also caused a profound decrease of MAP and HR. These results indicate that OFQ can inhibit cardiovascular activities which are not mediated by mu, delta and kappa receptors. POA may be one of the target areas of these inhibitory effects. PMID- 11498958 TI - Substance P- and 5-hydroxytryptamine-mediated depolarization in sympathetic ganglion neurons. AB - By means of intracellular recordings from sympathetic ganglion in vitro, the present study was to investigate whether the receptors of substance P (SP) and 5 hydroxytryptamine (5-HT) exist in the same neuron or separately in different neurons of guinea pig celiac ganglion (CG) and inferior mesenteric ganglion (IMG) and whether there are some interactions between the two transmitters. Of the 133 neurons of CG, 66 (49.6%) responded to both SP and 5-HT, 40 (30.1%) only to SP or 5-HT, 27 (20.3%) insensitive to both. The corresponding numbers of the corresponding groups of neurons of the 129 IMG neurons are 47 (36.4%), 65 (50.4%) and 17 (13.2%). Continuous superfusion of IMG with 5-HT did not affect SP depolarization, while continuous superfusion of IMG with SP did not affect 5-HT depolarization. The results indicate that SP receptor and 5-HT receptor may exist in the same neuron, and neither affects each other. PMID- 11498959 TI - Na+/Ca2+ exchange current in myocytes isolated from rat hypertrophied heart. AB - Effects of myocardial hypertrophy on the Na+/Ca2+ exchange current in isolated rat cardiac myocytes were investigated, using whole cell patch techniques. Goldblatt 2-kidney, one-clip technique was used to induce cardiomyocyte hypertrophy. The results demonstrated that the Ni(2+)-sensitive Na+/Ca2+ exchange current density was larger in hypertrophied cells than that in normal cells. At a holding potential of mV, the outward current densities were 1.53 +/- 0.31 pA/pF in normal cells and 2.62 +/- 0.53 pA/pF in hypertrophied cells (P < 0.01). At a holding potential of -100 mV, the inward current densities were 0.42 +/- 0.14 pA/pF in normal cells and 1.12 +/- 0.33 pA/pF in hypertrophied cells (P < 0.001). The results suggest that during myocardial hypertrophy the Na+/Ca2+ exchange current is significantly increased. PMID- 11498960 TI - [Effect of microinjection of taurine into anterior hypothalamic area on blood pressure in rats]. AB - The effect of taurine in anterior hypothalamic area (AHA) on blood pressure was investigated by means of microinjection and spectrophotofluorometry. The results are as follows. (1) Taurine microinjected into AHA decreased blood pressure markedly. (2) The depressor response of taurine in AHA could be antagonized by propranolol, but not by phentolamine. (3) After microinjection of taurine into AHA, the content of norepinephrine in hypothalamus increased significantly. The above results suggest that taurine in AHA attenuates blood pressure by increasing the content of norepinephrine, probably mediated by beta-adrenergic receptor. PMID- 11498961 TI - [Involvement of calcineurin-dependent signal pathway in the angiotensin II induced cardiac myocyte hypertrophy]. AB - The present study was undertaken to observe the role of calcineurin (CaN) dependent signaling pathway in the angiotensin II (Ang II)-induced cardiac myocyte hypertrophy. In cultured myocardial cells of neonatal rats, Ang II was used to stimulate hypertrophy and CaN-pathway blocked by CsA(an inhibitor of CaN). 3H-leucine incorporation, and activities of CaN, mitogen-activated protein kinase (MAPK) and protein kinase C (PKC) were investigated. The results showed that 3H-leucine incorporation of Ang II-stimulated myocardial cells was 46% higher than control (P < 0.01), which could be inhibited by CsA (0.5-5 micrograms/ml) and PD098059(an inhibitor of MAPK). CaN and PKC activities of Ang II-stimulated myocardial cells were 39% and 280% higher than control (P < 0.001) respectively, while no significant increase in MAPK activities was observed. CsA could reverse the increase of CaN activity, but had no effect on PKC. It is concluded that the CaN-dependent signaling pathway may play an important role in the development of the Ang II-induced cardiac myocyte hypertrophy. PMID- 11498963 TI - [Effect of stimulating the peripheral endings of transected dorsal root on discharges of neighbouring dorsal root in rats]. AB - The dorsal roots (DR) of L2-L3 segments on the left side in anesthetized rats (n = 28) were exposed and transected. After stimulating the peripheral endings of L2 DR with a train of square waves (0.8-1.2 mA, 100 Hz, 0.5 ms) for 2 s, the discharges at the distal end of transected L3 DR were recorded for 120 s. We found that the mean discharge frequency (MDF) of L3 DR was gradually increased with L2 DR stimulation. Time course analysis showed that the increase in MDF was linearly correlative to the number of stimuli, as well as to the levels of spontaneous discharge of L3 DR. The above results suggest that the transegmental information transmission between peripheral endings of primary sensory neurons may occur through a chemical non-synaptic way. PMID- 11498962 TI - Modulation of glycine-activated chloride currents by substance P in rat sacral dorsal commissural neurons. AB - The modulatory effect of substance P (SP) on strychnine-sensitive glycine (Gly) response was examined in neurons acutely dissociated from the rat sacral dorsal commissural nucleus (SDCN) using nystatin perforated patch recording configuration under voltage-clamp conditions. Application of SP potentiated 30 mumol/L Gly-activated chloride current (IGly) in a concentration-dependent manner over the range of 1 nmol/L to 1 mumol/L at a holding potential of -40 mV. SP neither changed the reversal potential of Gly response nor affected the affinity of Gly to its receptor. The SP potentiation effect could be blocked by spantide as well as a selective NK1 receptor antagonist, L-668, 169, but not by NK2 receptor antagonist, L-659, 877. The facilitatory action of SP on IGly could also be abolished by pretreatment with chelerythrine or KN-62 in different neurons, a finding suggesting that protein kinase C (PKC) or Ca2+/calmodulin-dependent protein kinase II (CaMK II) possibly contributes to an intracellular pathway of SP in the augmentation of IGly. The results imply that SP may suppress nociception in the spinal cord by potentiating Gly response. PMID- 11498964 TI - Interaction of cone signals on L-type horizontal cell in carp retina: experiments and modeling. AB - Intracellular recordings were made from the luminosity type cone driven horizontal cells (LHCs) in the isolated carp retina and model analysis was performed to investigate possible mechanisms underlying the interaction of different cone signals converging onto these cells. It was observed that a green background light enhanced the LHC's red response, and such enhancement was closely related to the activation of green cones. Model analysis showed that the activity of both glutamate- and GABA-related pathways were potentiated during green background illumination. GABA application did not abolish the response enhancement. It is speculated that the extent of the LHC's response enhancement may be determined by the balance of the increased activity between the feedforward and feedback pathways. PMID- 11498965 TI - [Role of red nucleus in inhibiting nociceptive responses of rat spindle afferent]. AB - Using extracellular recording techniques, the effect of excitation of red nucleus (RN) on C-responses of wide dynamic range (WDR) neurons and the roles of RN in the inhibitory effect of spindle afferent activated by succinycholine (SCH) on C responses of WDR neurons were studied. Stimulation of RN caused a significant reduction of nociceptive responses of WDR neurons, which was significantly attenuated by intravenous cyproheptadine and contralateral lesion of RN. These results suggest that the inhibitory effect on nociceptive responses of WDR neurons caused by RN stimulation was partly mediated by 5-HT and RN plays an important role in analgesia induced by the muscle spindle afferent activity. PMID- 11498966 TI - [Effect of ET-1 on intracellular free calcium in cultured neonatal myocardial cells]. AB - In this present study, the effects of ET-1 on intracellular free calcium concentration ([Ca2+]i) and the underlying mechanisms were investigated in cultured neonatal rat myocardial cells loaded with fura-2/AM. The results are as follows. ET-1 induced an increase of [Ca2+]i in a dose-dependent manner, which consisted of a transient and sustained phase. BQ123, a selective ETA receptor antagonist, blocked the ET-1 induced [Ca2+]i responses, suggesting that these responses were mediated by ETA receptors. After removal of extracellular Ca2+, ET 1 induced the transient increase of [Ca2+]i without the sustained change. Protein kinase C (PKC) agonist PMA attenuated the ET-1 induced transient [Ca2+]i increase. Amiloride and nifedipine did not block the [Ca2+]i change induced by ET 1. After pretreatment of myocardial cells with pertussis toxin, ET-1 also induced the transient increase of [Ca2+]i but did not affect the sustained increase. These results suggest that the transient [Ca2+]i increase may involve pertussis toxin-insensitive G protein and the sustained one may be caused by extracellular calcium influx, in which pertussis toxin sensitive G protein is involved. Furthermore, PKC, but not Na+/H+ exchange, plays an important role in these effects. PMID- 11498967 TI - [Inhibitory effects of antisense oligonucleotides targeting mitogen-activated protein kinase (MAPK) mRNA on neonatal rat cardiac fibroblast proliferation induced by Ang II and EGF]. AB - In the present study, the antisense oligodeoxynucleotide (ODN) approach was used to investigate whether mitogen-activated protein kinase (MAPK) is necessary for the proliferation response in neonatal rat cardiac fibroblast (FB) induced by angiotensin II (AngII) or epidermal growth factor (EGF), the proximal cytosolic signal transduction pathways which are quite different processes. A phosphorothioate-protected 17-mer directed against the initiation of translation sites P42 MAPK mRNA was introduced into FB by liposomal transfection. The results showed that (1) after a 24 h treatment with AngII or EGF (all 10(-8) mol/L), the FB numbers were increased by 39% and 68%, while the rate of DNA synthesis increased by 60% and 102%, respectively. (2) Following 5 min or 10 min stimulation with AngII or EGF, MAPK activity ([gamma-32P] ATP incorporation) increased by 202% and 305%, and phospho-MAPK protein content increased by 545% and 646% correspondingly. (3) As compared with lipofectin + AngII/EGF control, after pretreatment with MAPK antisense ODN, the MAPK protein expression was inhibited significantly; the rate of DNA synthesis of FB induced by AngII or EGF was reduced by 53% and 46%, cell numbers by 38% and 44%, respectively. Meanwhile, MAPK activity was decreased by 74.2% and 65.9%, phospho-MAPK protein content by 85% and 90%. The sense or random ODN has not much effect on them. Consequently, it can be concluded that (1) MAPK activity is essential in the event of involving FB proliferation response reduced by AngII and EGF, and (2) FB proliferation response could be inhibited by the MAPK antisense ODN through depletion of MAPK. PMID- 11498968 TI - [Hypoxia-induced increase in nerve activity of rabbit carotid body mediated by noradrenaline]. AB - The responses of 39 hypoxia-sensitive units of chemoreceptive afferent in sinus nerve to noradrenaline (NA) and its antagonist were recorded in carotid body sinus nerve preparations from 30 rabbits. The results are as follows. (1) Discharges of the units increased from 0.13 +/- 0.06 to 0.25 +/- 0.12 imp/s (P < 0.001, n = 19) upon lowering PO2 of modified Tyrode solution. (2) Adding NA (10( 6) mol/L) to the perfusate led to an increase in the unit discharge from 0.14 +/- 0.08 to 0.23 +/- 0.13 imp/s (P < 0.01, n = 19). (3) Prazosin (10(-6) mol/L) did not alter the basal frequency of chemosensory unit discharges under normoxic conditions (P > 0.05, n = 4). (4) Yohimbine (10(-6) mol/L) did not alter the basal frequency of chemosensory unit discharges under normoxic conditions (P > 0.05, n = 6). (5) Chemosensory responses to hypoxia were not altered after pretreatment with prazosin. (6) Chemosensory responses to hypoxia were inhibited by pretreatment with yohimbine. The present results suggest that (1) NA is not mainly concerned with spontaneous discharges of chemoreceptor sensitive to hypoxia, but does elicit an increase in spontaneous discharges, and (2) the increase of chemosensory unit discharges produced by hypoxia can be inhibited by yohimbine. It is likely that the excitatory action of hypoxia on chemoreceptive process is mediated by NA. PMID- 11498969 TI - [Effect of ET-1 antisense oligodeoxynucleotide on the hemodynamics of normal and experimental hypertensive rats]. AB - It was previously found that a phosphorothioated antisense oligodeoxynucleotide (ETASODN) significantly inhibits production of endothelin-1 (ET-1). The purpose of the present study was to determine whether intracerebroventricular injection of ETASODN targeted to prepro-ET-1 is capable of exerting the same preventing effect on the aorta narrowing of experimentally modeled hypertensive rats. Radioimmunoassay showed that ET-1 level in the brain stem of hypertensive rats was significantly elevated. In addition to down-regulating the ET-1 level, astisense could also reduce mean arterial pressure (MAP), heart rate and LVSP in model rats. The antisense also down-regulated the ET-1 level in hypothalamus and brain stem, reducing MAP in normal control rats. After treatment with the antisense, the value of delta MAP was markedly lowered in experimental hypertensive rats as compared to the control ones. Thus it appears that (1) ET-1 might play an important role in central cardiovascular regulation in rats and (2) antisense ETASODN might be used in treatment of hypertension via inhibiting ET-1 production. PMID- 11498970 TI - [Vasoactive intestinal peptide and epidermal growth factor upregulates bcl-2 gene expression in bronchial epithelial cells]. AB - Expression of protooncogenes bcl-2 and c-myc in cultured rabbit bronchial epithelial cell (BEC) was investigated in order to shed some light on genetic mechanisms underlying the protective antioxidant effect of pulmonary regulatory peptides, vasoactive intestinal peptide (VIP) and epidermal growth factor (EGF). Effects of these peptides and heat stress (HS) on expression of these genes were also studied. Total RNA was extracted from BEC. Bcl-2 mRNA and c-myc mRNA were cloned with the method of RT-PCR. GAPDH mRNA was used as internal control. The products of RT-PCR were separated with electrophoresis in 2% agarose gels. A computer image treating system (Stratagene eagleeye II) was used to identify the specific band and evaluate the density. The product bands of target genes bcl-2 were checked with Southern blot and oligoneucleotides probe hybridyzation. The results show: (1) a low level of bcl-2 and c-myc gene transcription occur in BEC at the resting state; (2) both VIP and EGF could promote bcl-2 and c-myc transcription, but no significant change could be found in the HS group; (3) there was a close correlation between bcl-2 and c-myc transcription (r = 0.98. P < 0.01). The above results indicate that VIP and EGF can improve the antioxidant effect of BEC by upregulating bcl-2 gene expression potently modulated by c-myc protein. PMID- 11498971 TI - Characteristics of apyrase (EC 3.6.1.5) on cultured bovine endocardial endothelial cells. AB - Apyrase activities in some tissues and cells, such as peripheral vascular endothelial cells, have been reported, but these in endocardium endothelial cells have not been reported. The present study was to characterise the properties of bovine endocardium endothelial cells (BEEC)-associated apyrase. Apyrase activity was assayed by inorganic phosphate release, which could be inhibited concentration-dependently by NaN3, an apyrase inhibitor. NaF (20 mmol/L), another inhibitor of apyrase, also markedly inhibited the activity. EDTA or EGTA (1 mmol/L) could also inhibit the activity completely. However, the inhibitor for Na+/K(+)-ATPase, ouabain (3 mmol/L) did not affect the enzyme activity. BEEC apyrase activity was dependent on divalent cations (Ca2+ or Mg2+) and pH value. PMID- 11498972 TI - [Antagonistic effects of melatonin on glutamate-induced neurotoxicity in rat hippocampal neurons]. AB - The effects of glutamate (Glu) and melatonin (MEL) on the evoked populaion spike (PS) following stimulation of the schaffer collateral fiber were studied by extracellular recording technique in perfused slices of rat hippocampus: 5.0 mmol/L Glu decreased the PS peak values to 4.1% of control. This effect of Glu (5.0 mmol/L) on PS peak values was changed to 14.7%, 105.2% and 24.3% of control by 0.4, 0.5, 0.6 mumol/L of MEL. But when the interaction between MEL (0.5 mumol/L) and Glu (5.0 mmol/L) was entirely suppressed by CDP (0.5 mumol/L), the PS values reduced to zero. These results suggest that the inhibitory action of MEL on Glu-induced neurotoxicity may be mediated by 5-HT receptors. PMID- 11498973 TI - [The enhancing effects of arginine-vasopressin on antibody level and lymphocyte proliferation of rats]. AB - The effects of arginine-vasopressin (AVP) on some immune parameters of rats were studied by using icv of AVP. The hemolysin to sheep red blood cell and IgG production were enhanced by icv of 100, 800 ng AVP. Both the actions were partly blocked by icv preinjection of the V1 receptor antagonist DPAVP. It was also found that the NE and corticosterone levels in the serum of rats were significantly reduced after icv of AVP. The above results suggest that AVP executes immunofunctions through V1 receptors in the paraventricular nucleus of the brain. Considering our previous results, the AVP enhanced immunofunction may be related to its inhibition of corticotropin releasing factor release or the excitatory activity of sympathetic nerves. PMID- 11498974 TI - [Effects of stimulation of dorsal medial area and ventral medial area of nucleus facialis on myoelectric activities of genioglossus and phrenic muscle]. AB - Effects of electrical and chemical stimulation of the dorsal and ventral medial area of nucleus facialis (dMNF and vMNF) on myoelectric activities of genioglossus and phrenic muscle were observed in 44 urethane-anaesthetized and vagotomized rabbits. The results are as follows. (1) Long train electrical stimulation at the dMNF facilitated myoelectric activities of genioglossus and phrenic muscle. (2) Short train electrical stimulation of dMNF, when delivered in the inspiratory phase, caused prolongation of myoelectric activities of genioglossus and phrenic muscle; but when delivered in the expiratory phase, myoelectric activities of genioglossus and phrenic muscle were shortened. (3) Long train electrical stimulation at the vMNF inhibited myoelectric activities of genioglossus and phrenic muscle. (4) Short train electrical stimulation of vMNF caused shortening of myoelectric activities of genioglossus and phrenic muscle, when delivered in the inspiratory phase, but a prolongation when delivered in the expiratory phase. (5) The effects of microinjection of glutamate to dMNF and vMNF were similar to those of electrical stimulation. The above results suggest that both dMNF and vMNF participate in the regulation of the myoelectric activities of genioglossus and the upper airway resistance. PMID- 11498975 TI - [Melatonin decreases the release of amino acid transmitters in hypoxic rat cortical slice]. AB - To study the effect of melatonin on changes in amino acid release following hypoxic stimulation, a RP-HPLC fluorometric method was used to measure the content of amino acids in the culture medium of rat brain slices. Hypoxic condition was instituted by a gas of 91.6% N2 and 8.4% O2. The results showed that the contents of amino acids, including aspartate, glutamate, glutamine, glycine, taurine and GABA, in the medium were significantly increased to 240.4%, 334.3%, 200.6%, 210.4%, 168.6% and 263.9%, respectively (P < 0.01 vs control) by 30 min hypoxic incubation. Melatonin can significantly reduce the increase of aspartate and glutamate, to 55.1% and 57.0% of the hypoxic group. PMID- 11498976 TI - [Electrophysiological and morphological characteristics of the visceral nociceptive neurons in cortex S II area of cats]. AB - Electrophysiological and morphological properties of the visceral nociceptive neurons (VNNs) in the secondary somatosensory cortex (SII) area of 16 cats were investigated with intracellular recording and labeling method. Electrophysiological properties of a total of 251 neurons were identified, 109 of which were induced to discharge by stimulating the greater splanchnic nerve and therefore were simply designated as VNNs. Various patterns of the response could be recorded: excitatory (38.53%), inhibitory (42.20%) and excitato-inhibitory (19.31%). Following acquisition of electrophysiological data, neurobiotin was injected into 21 cells by electrophoresis to show their morphology and distribution in the cerebral cortex. It was found that excitatory VNNs were pyramidal neurons predominantly, whereas more inhibitory VNNs were stellate neurons. PMID- 11498977 TI - [Increased release of orphanin FQ (OFQ) in brain of chronic morphine tolerant rats]. AB - The changes in OFQ-immunoreactivity (OFQ-ir) content and release of cerebroventricular perfusate, periaqueductal gray (PAG) and amygadala of chronic morphine tolerant rats were measured with radioimmunoassay (RIA). The results indicate: (1) chronic morphine tolerance was achieved in rats by injecting increasing doses of morphine (10, 20, 40, 50, 60 mg/kg, s.c., tid). The content of OFQ-ir in cerebroventricular perfusate in the rats of normal saline (NS) group remained at a steady level during the injections from d 1 to d 5, while in the morphine-treated group of rats the content of OFQ-ir showed 25% increase (P < 0.05 vs NS) and 52% increase (P < 0.01 vs NS) after 3 and 5 days' morphine injection respectively. (2) The content of OFQ-ir in PAG of rats receiving morphine injection for 1, 3 and 5 days showed respectively increases of 17% (P < 0.05), 48% (P < 0.05) and 80% (P < 0.01) against the NS group. (3) The content of OFQ-ir in rat amygdala receiving 1 day injection of morphine showed a 8% decrease, which was not significantly different from NS group. However, there was a 36% and 55% (P < 0.05) increase respectively after injection for 3 and 5 days. It is suggested that in the later stage of chronic morphine treatment, large amount of OFQ was released from rat brain to antagonize the effect of opioids, which may play an important role in the development of morphine tolerance. PMID- 11498978 TI - Effect of removal of external calcium on phosphoinositide hydrolysis in cultured myotubes of embryonic chicken. AB - The effect of removal of external Ca2+ on phosphoinositide hydrolysis was investigated in cultured myotubes from 9-day-old Leghorn embryonic chicken. In the myotubes exposed to Ca(2+)-free Ringer's solution, the turnover of phosphoinositide was exponentially decreased with a time constant of about 26 min. In the presence of external Ca2+, the hydrolysis of phosphoinositide was significantly increased by exposure to 80 mmol/L K+ solution. After removal of external Ca2+, 80 mmol/L K+ exposure caused a slight decrease of phosphoinositides hydrolysis in comparison with the control (normal Ringer). It is indicated that hydrolysis of phosphoinositide in cultured myotubes can be enhanced by high K+ exposure. External Ca2+ is essential for this effect, which is different from mature muscle fibres. PMID- 11498979 TI - [Effect of selective lesions in hypothalamus on bilateral synchronized bursts of rat oxytocin neurons]. AB - In order to clarify the location of the center for synchronized milk-ejection bursts of magnocellular oxytocin neurons in the hypothalamus, the bursts of these neurons were recorded extracellularly in lactating rats with selectively-cutting lesions of the middle brain or hypothalamus. Results showed that unilateral transection of the middle midbrain above the ventral tegmentum did not block the synchronized bursts on both sides; however, the synchronized bursts disappeared after unilateral transection through the middle of the medial hypothalamus. These results suggest that the area from the middle part of the midbrain to that of the hypothalamus does play a crucial role in the synchronized milk-ejection burst. PMID- 11498980 TI - [Role of caudal pressor area of medulla oblongata in vasomotor tone of peripheral vessels]. AB - To study the role of the caudal pressor area (CPA) on the ventral surface of medulla oblongata in vasotonia of the skeletal muscles and kidneys, perfusion pressures of vessels in skeletal muscles and kidneys were recorded to observe effects of microinjection of L-glutamate (L-glu) into CPA. The results are as follows. L-glu induced a significant increase of artery pressure (AP), perfusion pressure of muscles (PPm) and perfusion pressure of kidney (PPk), which was markedly attenuated by prior injection of phentolamine or propranolol to the bilateral pressor area. These results indicate that the vascular roles of CPA were mainly mediated via alpha- and/or beta-receptors. PMID- 11498981 TI - [Changes in vasopressin V1 receptor mRNA level in rats with high blood pressure induced by chronic stress]. AB - Using reverse transcription polymerase chain reaction (RT-PCR) combined with Southern blotting hybridization technique, the AVP V1 receptor mRNA was found to be widely distributed in central neuronal system and other tissues of rats, such as cerebral cortex, hypothalamus, medulla, liver and kidney. The receptor mRNA levels in the cortex, hypothalamus and medulla were decreased significantly in chronically stressed rats, as compared with normal controls (cortex: P < 0.05; hypothalamus: P < 0.01; medulla: P < 0.001). But no significant change was observed in tissues of heart, liver and kidney (heart: P > 0.05; liver: P > 0.05; kidney: P > 0.05). These results suggest that chronic stress may lead to a decrease of AVP V1 receptor density in the CNS as a result of decreased synthesis. PMID- 11498982 TI - [Time effect of restraint stress-induced suppression of lymphocyte transformation]. AB - In the present study, restraint, which caused no direct tissue damage, was selected as a method of stress to investigate time effect of stress-induced suppression of lymphocyte transformation in rats. The lymph node and spleen extract or the serum from rats restrained for 3, 6, 12, 18 h or 12, 24, 48, 72, 96 h after restraint, with concanavalin A (Con A), was added to the lymphocyte suspension of rat mesenteric lymph node and was incubated for 72 h. The optical density (OD) value indicating the level of lymphocyte transformation was determined by colorimetric assay of methylthiazol tetrazolium bromide (MTT). The results are as follows. (1) The lymph node and spleen extract or the serum from the rats restrained for 3 or 6 h did not markedly affect the lymphocyte transformation. (2) The lymph node and spleen extract or the serum from the rats restrained for 12 h or 18 h significantly inhibited the lymphocyte transformation and stronger inhibition occurred at 18 h. (3) Up to 96 h after restraint, the suppressive effect of the lymph node or spleen extract on the lymphocyte transformation still existed, but the suppression of the serum on the lymphocyte transformation almost disappeared. These results suggest that the restraint of rats may cause lymphoid tissues to produce some immune suppressive protein of stress, which is released into blood leading to a generalized suppression of cellular immunity. PMID- 11498983 TI - [Role of c-fos gene in the pulmonary surfactant synthesis of cultured alveolar type II cells induced by endothelin-1]. AB - The effects of endothelin-1 (ET-1) on pulmonary surfactant (PS) synthesis of cultured alveolar type II cells (AT II) were observed. The role of c-fos gene in cellular signal transduction of ET-1 was studied by antisense technology. The results showed that: (1) ET-1 enhanced [3H] choline incorporation into AT II cells in a dose-dependent manner. (2) Protein kinase (PKC) activator PMA increased [3H] choline incorporation into AT II cells, while PKC inhibitor H7 inhibited the stimulating effect of ET-1. (3) Both ET-1 and PMA could increase the level of c-Fos protein, and H7 and c-fos antisense oligonucleotides (AS ODN) could inhibit the effects induced by ET-1 on Fos protein expression and [3H] choline incorporation. (4) The release of lactic dehydrogenase (LDH) was not different among control, ET-1, antisense oligonucleotides and sense oligonucleotides groups. The above results demonstrated that ET-1 can enhance PS synthesis of AT II cells and ET-1 stimulating the expression of c-fos gene mediated by PKC is a major signal transduction pathway of modulating PS synthesis. PMID- 11498984 TI - [Nestin expression during P19 neuron differentiation]. AB - Mouse nestin, an intermediate filament gene, is transiently expressed during the development of the central nervous system. In order to find the clue of its function during neural development, we tried to find out the gene expression pattern during the neuronal differentiation of P19 EC cells induced RA. RT-PCR showed that nestin was transiently expressed during P19 neuron differentiation, with a peak at day 4 of this process. However, BMP4, a neural precursor cell marker, was transiently expressed with its highest level at day 6, while NF160 kD a terminal differentiated neuronal marker, was increasingly expressed during the whole process. These results implied that nestin might play some roles during the process of neural progenitor cells differentiating into neural precursor cells. Moreover, immunostaining showed that nestin was located in the neurite and the growth cone of the P19 neuron, suggesting that nestin might be also involved in the process of the establishment of neural connection. PMID- 11498985 TI - [Effect of beta-endorphin on plasma sialic acid level in rats and the relationship with the immune function]. AB - With the use of beta-EP microinjection into the central nervous system, spectrophotometrical analysis and immunochemical method we studied the effect of beta-Endorphin (beta-EP) on plasma sialic acid (SA) level. The results indicate: (1) The plasma SA level was significantly decreased after microinjection of beta EP into the lateral ventricle (LV); (2) When beta-EP was microinjection into nucleus arcuatus (ARC), the plasma SA level was significantly decreased (P < 0.05). This effect can be blocked by i.v. atropine or vagotomy; (3) Microinjection of beta-EP into ARC can cause the increase of CD3, CD4 and CD4/CD8, while CD8 was decreased. The above results suggest that the possible mechanism underlying the decrease of plasma SA level is mediated by cholinergic muscarinic receptor, and the cellular immune function is enhanced. PMID- 11498986 TI - [Beta-endorphin involved in the regulation of humoral immune function of rats during acute hypoxia]. AB - In order to investigate the role of beta-endorphin in the regulation of humoral immune function of rats during acute hypoxia, the effects of beta-endorphin on mitogen-induced spleen lymphocyte DNA synthesis and hemolysin formation as well as IgG production to chicken egg albumen were observed. It was found that after rats received icv injection of beta-endorphin (1 ng/rat), the T-lymphocyte DNA content, the hemolysin-forming capacity of SRBC-sensitized rats and the IgG level were reduced significantly compared with the control (icv normal saline). Similar suppressive effects on immune function could be found after rats were exposed to 7 km altitude in a hypobaric chamber for 48 h. Pretreatment with icv injection of naltrexone, the immunosuppressive effects of acute hypoxia were partially blocked. Icv administration of beta-endorphin produced an increase in splenic cathcholamines, similar to those of hypoxia treated group (7 km for 12 h). All the above findings suggest that beta-endorphin may modulate the immune response to hypoxia stress via opioid receptor and its inhibitory action may be mediated by activating sympathetic nervous system. PMID- 11498987 TI - Intracarotid injection of endothelin-1 facilitates the activity of rostral ventrolateral medullary neurons via area postrema in rats. AB - To observe the effect of intracarotid administration of endothelin (ET-1) on electrical activity of neurons within rostral ventrolateral medulla (RVLM) region, 87 spontaneous active units were extracellularly recorded in 35 Sprague Dawley rats with sino-aortic denervation. The results obtained are as follows. (1) Intracarotid administration of ET-1 (0.3 nmol/kg) increased the discharge firing rate from 17.8 +/- 1.5 to 20.9 +/- 1.4 spikes/s (P < 0.01) in 30 out of 36 RVLM neurons, while blood pressure and heart rate had no significant change. (2) BQ-123 (0.67 nmol/kg), a selective ETA blocker, completely blocked the facilitatory effects of ET-1 in 11 out of 14 units. (3) In 10 out of 11 units, glibenclamide (3.3 nmol/kg), a blocker of ATP-sensitive potassium channel, had no effect on the action of ET-1. (4) After ablation of area postrema (AP), the facilitatory action of intracarotid administration of ET-1 on 19 units of RVLM was abolished, while in 7 units of sham ablation animals the response of neurons to ET-1 remained unchanged. Taken together, intracarotid-administered ET-1 may act on the ETA receptors in neurons of AP, thereby resulting in the facilitating effect on RVLM neurons through the efferent projection of AP. PMID- 11498988 TI - [Atrioventricular conduction changes in acute ischemic and reperfused coronary artery of cats]. AB - Acute inferior wall infarction is frequently complicated by conduction disturbance, but the relationship between ischemia and atrioventricular (AV) conduction disturbance is not well understood. The present experiments were made on cats with heart efferent completely denervated. A, H and V waves were picked up by a template-machine algorithm from His bundle electrogram, AA interval (cardiac cycle length), AH interval (AV conduction time), HV and AV intervals were measured. Of 20 cats, 14 showed prolongation of AH interval (Group A), while not in 6 cats (Group B) after ligation of right coronary artery. A fast pacing of right atrium (AA interval = 267 ms) was produced in Group B and AH interval was also prolonged after ligation of the artery. By multiple-step pacing, the curve of AV nodal function shifted to the right, AH interval which induced AV nodal block was prolonged after ischemia. After reperfusion, AH interval was shortened both in Group A and B. The above results show that myocardial ischemia may cause abnormality of AV conduction and accommodation function. The abnormality appears to be a potential danger which will induce AV conduction block during tachycardia. PMID- 11498989 TI - [Light and electron microscopic observation of zinc distribution in the carp retina]. AB - Localization of zinc in the carp retina in both light and dark adaptation was investigated using neo-Timm sulfate silver method. The results showed that in light adaptation, zinc was preponderantly present in the outer plexiform layer (OPL). Both terminals of zinc containing photoreceptors and dendrites of bipolar cells usually extend into zinc staining areas in OPL. In dark adaptation, zinc staining in retina is blanched or only appeared in cytoplasmic bodies of few cones. There is no significant difference between dark and light adaptation in the zinc distribution in bipolar, amacrine and ganglion cells. The fact that zinc is perched in the carp retina between photoreceptors and second-order neurons of the visual pathways in OPL in light adaptation and significantly released in dark adaptation suggests that zinc might play some modulatory roles in the visual signal transduction. PMID- 11498990 TI - [Expression of cyclin dependent kinase inhibitor p27 during proliferation in vascular smooth muscle cell]. AB - This study was to investigate cell cycle distribution of the vascular smooth muscle cells (VSMCs) and negative regulator of cell proliferation p27 expression caused by platelet derived growth factor BB (PDGF-BB), angiotensin II (Ang II) and arginine vasopressin (AVP). Deprived of fotal calf serum for 48 h, cultured VSMCs in quiescent condition were collected at different times after stimulation of Ang II, AVP and PDGF-BB. Cell cycle distribution and p27 expression were determined with a flow cytometer. The results showed that the protein content of VSMCs was significantly increased (43.6%) by Ang II as a result of hypertrophy, but Ang II did not lead to downregulation of p27. AVP could downregulate p27 slightly. PDGF could inhibit p27 expression significantly and cause VSMCs hyperplasia. These results suggest that the progression of VSMCs through G1 to S phase might be brought out by the inhibition of p27 during proliferation. PMID- 11498991 TI - [Expression of tissue factor of astrocytes and its signal transductional pathways]. AB - The aim of this study was to observe the expression of tissue factor (TF) of astrocytes in basic culture medium and under the condition stimulated by thrombin and to explore the relevant signal transduction pathways. The results showed that 4-bromo calcium ionophore (A23187) and phorbol 12-myristate 13-acetate (PMA) enhanced significantly the TF expression of astrocytes, but the expression was decreased markedly by trifluoperazine (TFP) and 1-(5-isoquinolinyl sulfonyl)-3 methyl-piperazine (H7) in the basic medium. Thrombin increased significantly the TF expression of astrocytes, which was obviously inhibited by TFP and H7. The results above indicate that astrocytes can express TF activity in the basic medium, which is promoted by thrombin, probably through some pathways involving Ca2+/CaM and protein kinase C (PKC). PMID- 11498992 TI - [Effects of monocarboxylic acid derivatives on cardiac ventricular CFTR Cl- channels in guinea pig]. AB - Using the whole-cell recording technique, the effects of monocarboxylic acid derivatives on cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel were examined in guinea pig ventricular myocytes. Anthracene-9-carboxylic acid (9-AC) added to the bath solution further enhanced the outward component of isoproterenol-induced currents in a reversible manner, whereas 5-nitro-2-(3 phenylpropylamino) benzoic acid (NPPB) or diphenylamine-2-carboxylic acid (DPC) induced a biphasic effect on the currents. Either NPPB or DPC first produced a transient increase in the outward component of current before ensuing inhibition. Intracellular NPPB was found to potentiate isoproterenol-activated currents. It is concluded that these monocarboxylic acid derivatives have different binding sites in cardiac ventricular myocytes, which might partially account for the varied effects in blocking anion channels. PMID- 11498993 TI - [Bradykinin inhibits carotid sinus baroreflex in anesthetized rats]. AB - The effects of bradykinin (BK) on the carotid baroreflex were examined in 36 anesthetized rats with isolated carotid sinus perfusion. The results obtained are as follows. (1) By perfusing the isolated carotid sinus with BK (1.0 mumol/L), the functional curve of baroreflex was shifted to the right and upward, its peak slope (PS) decreasing from 0.44 +/- 0.14 to 0.31 +/- 0.01 kPa (P < 0.01) and the reflex decrease in mean arterial pressure (RD) was lowered from 6.85 +/- 0.18 to 4.46 +/- 0.16 kPa (P < 0.05), while the threshold pressure (TP), equilibrium pressure (EP) and saturation pressure (SP) were significantly enhanced from 7.76 +/- 0.20 to 10.04 +/- 0.09 kPa (P < 0.001), 12.72 +/- 0.29 to 13.74 +/- 0.31 kPa (P < 0.05) and 23.28 +/- 0.24 to 25.31 +/- 0.20 kPa (P < 0.01), respectively. Among the functional parameters of carotid baroreflex, the changes in RD, PS and TP induced by BK were dose-dependent. (2) By pretreatment with indomethacin (10 mumol/L), an inhibitor of prostaglandin synthesis, the above-mentioned effects of BK on carotid baroreflex were not affected. (3) Preperfusion with an inhibitor of NO synthase L-NAME (100 mumol/L) could completely eliminate the effects of BK. (4) Pretreatment with angiotensin-converting enzyme inhibitor captopril (20 mumol/L) could potentiate the effects of BK. Taken together, it is indicated that BK may cause vascular endothelium to release NO, thereby inhibiting the carotid baroreflex. PMID- 11498994 TI - [Ethanol-induced gastric mucosal injury and the protection of taurine against the injury in rats]. AB - 40 SD rats were randomly divided into control group, ethanol-induced injury group and taurine protective group for the measurement of the contents of endothelin (ET), nitric oxide synthetase (NOS), somatostatin (SS) and vasoactive intestinal peptide (VIP). The results are as follows. The gastric mucosa was damaged by 75% ethanol, being aggravated with prolonged application time, as shown by a significant increase of the ET content and decreases of the contents of NOS, VIP and SS in the gastric mucosa. In the taurine group, decrease of ET content and increase of the contents of NOS, SS and VIP in the gastric mucosa were observed. Apparently, the changes in ET, NOS, SS and VIP contents of gastric mucosa serve as reliable indices in the pathogenesis by ethanol and protective effect by taurine. PMID- 11498995 TI - [The role of endogenous CO in the regulation of endothelin-induced VSMC proliferation and MAPK activity]. AB - Heme oxygenase (HO) is a rate-limiting enzyme of heme degradation, which converts the cellular heme to bilirubin and carbon monoxide (CO). Recently it is suggested that endogenous CO plays an important role in regulating vascular tone under both physiological and pathological conditions, but it is not clear whether endogenous HO/CO system regulates vascular smooth muscle cell (VSMC) proliferation. In the present study, VMSC 3H-TdR incorporation, mitogen-activated protein kinase (MAPK) activity, HO activity and CO release were determined to study the role of endogenous HO/CO system in regulating the VSMC proliferation induced by endothelin-1 (ET-1) in a cultured system. The results showed that ET-1 increased VSMC 3H-TdR incorporation, MAPK activity, HO activity, and CO release were up regulated. Pretreatment of HO inhibitor, zinc protoporphyrin-9 (ZnPP-9), increased the ET-1-induced VSMC 3H-TdR incorporation and MAPK activity by 31.8% and 36.6% (P < 0.01, respectively), whereas pretreatment of heme-L-lysinate (HLL), a HO substrate, inhibited these activities. This study demonstrated that up-regulation of VSMC endogenous HO represents a cellular protective response to stress or injury. Inhibition of HO may enhance VSMC proliferation induced by ET-1 in vitro, suggesting that endogenous HO/CO system may be directly involved in the regulation of VSMC proliferation through MAPK signaling pathway. PMID- 11498996 TI - [Electrophysiological effects of agmatine on guinea pig papillary muscles in vitro]. AB - The cardiac electrophysiological effects of agmatine (AGM) were examined in guinea pig papillary muscle using intracellular microelectrode technique. The results obtained are as follows. (1) Duration of action potential (APD) in normal papillary muscles were decreased by AGM in a concentration-dependent manner. (2) In partially depolarized papillary muscles, amplitute of action potential, overshoot, maximal velocity of phase 0 depolarization and APD were depressed by AGM. (3) Pretreatment with NG-nitro-L-arginine methyl ester (L-NAME, 0.5 mmol/L) did not affect the above effects of AGM (1 mmol/L) on papillary muscles. (4) The effects of AGM (1 mmol/L) could be blocked completely by pretreatment with idazoxan (0.1 mmol/L), an alpha 2-adrenoceptor (alpha 2-AR) and imidazoline receptor (IR) antagonist. All these results indicate that the effects of AGM on papillary muscles are likely due to a decrease of intracellular calcium mediated by alpha 2-AR and IR. PMID- 11498997 TI - [Effect of enkephalin on glial neurotrophic function related to the reduction of nitric oxide production]. AB - The effect of methionine-enkephalin (ME) on glial neurotrophic function was studied in rat cortical glial-neuronal coculture. The results showed that ME treated glia enhanced neuronal survival by 28% (P < 0.05), and increased total neurite length per neuron by 11% (P < 0.05), while the expression of GAP-43 mRNA by 26% (P < 0.05). The effect of ME on NO production of glia was also studied in rat cortical glial culture. The result showed that different concentrations of ME (10(-12), 10(-10), 10(-8), 10(-6) mol/L) inhibited glial NO production. All the above show that the increase of glial neurotrophic ability by ME may be due to the inhibition of glial NO production. PMID- 11498998 TI - [The inhibitory effect of amygdaloid stimulation on the "on-off" response of medial geniculate body neurons in rabbits]. AB - Experiments were performed on 23 New Zealand rabbits immobilized with Flaxedil. The characteristics of on-off responses of medial geniculate body (MGB) neurons to tone bursts of varying frequency, intensity and duration were affected by lateral amygdaloid nucleus (LAm) stimulation, which was in form of altering the pattern of discharge or complete suppression. The effect also varied with the changes of the interval between amygdaloid stimulation and the tone bursts. The results of the present work show that LAm could modulate temporal processing of auditory information. PMID- 11498999 TI - [Alteration of ryanodine receptors in cardiac sarcoplasmic reticulum and nuclear envelope of rats during sepsis]. AB - To investigate changes of ryanodine receptors in the sarcoplasmic reticulum (SR) and the nuclear envelope (NE) of rat cardiac myocytes during sepsis induced by cecal ligation and puncture (CLP), myocardial SR and NE were fractionated with density gradient centrifugation and the characteristic of ryanodine receptor was assayed with a method of radioreceptor binding assay. The result showed that Bmax of ryanodine receptors in cardiac SR was increased by 23% during early sepsis (9 h after CLP), but decreased by 38% during late sepsis (18 h after CLP). Bmax of ryanodine receptors in cardiac NE, on the other hand, was increased by 100% and 160% during early and late sepsis respectively. Kd of ryanodine binding to SR and NE remained unchanged during sepsis. These results demonstrated up-regulation of ryanodine receptors in SR occurred during early sepsis and down-regulation of these receptors in SR occurred during late sepsis, while up-regulation of ryanodine receptors in NE occurred during both the early and the late sepsis. PMID- 11499001 TI - [Biological activity of recombinant human hepatopoietin]. AB - We examined the effects of rhHPO on the cell growth and DNA synthesis of both rat primarily cultured hepatocytes and hepatic carcinoma cell line in vitro by MTS and 3H-TdR in corporation methods. It was indicated that rhHPO is an important stimulating factor of regeneration, which may be developed as a potential drug for the treatment of severe hepatic diseases. We also found an inhibitory effect of rhHPO on the DNA synthesis of lung cancer cell lines GLC-82 in vitro, which might provide a valuable indicator for the study of its specificity and mechanisms. PMID- 11499000 TI - Detection of release of aspartate and glutamate in cat spinal dorsal horn following noxious stimulation of cutaneous and muscular afferent fibers using microdialysis. AB - To investigate the possible mechanisms underlying the difference of NMDA and non NMDA receptors in spinal nociception originating in skin and muscle, release of aspartate (Asp) and glutamate (Glu) in the spinal dorsal horn was detected by stimulation of cutaneous and muscular nerves in cats using microdialysis technique. Asp and Glu were increased respectively by (323 +/- 55)% and (169 +/- 16)% following stimulation of cutaneous nerve, but by (150 +/- 16)% and (218 +/- 42)% respectively following stimulation of muscular nerve. Asp increase was approximately three times higher than that of Glu following cutaneous nerve stimulation (P < 0.01), while Glu increase was approximately twice as high as that of Asp following muscular nerve-stimulation (P < 0.05). It is likely that nociceptive cutaneous and muscular inputs preferentially elicite release of Asp and Glu respectively, resulting in a functional differentiation of NMDA and non NMDA receptor in the mediation of different nociceptive information. PMID- 11499002 TI - [Vasoactive intestinal polypeptide induced prolactin release is model dependent in free-moving rats]. AB - Our previous experiments in vitro showed that the stimulating effects of vasoactive intestinal polypeptide (VIP) on pituitary (PRL) depended on the endocrinal status of the animals. The present investigation was to determine whether the effect of VIP varied in vivo with changes of different physiological conditions. For infusion of VIP (5 micrograms/100 g body weight) and collection of blood sample, all the animals were cannulated with silicon tube into jugular vein 2-3 d before the experiments. The results showed that VIP concentration in blood was increased rapidly after the infusion (maximum: 21.32 +/- 2.33 ng/ml at 10 min and lasting more than 30 min). The concentration of PRL in blood of all the animals tested was increased significantly (P < 0.05) after VIP infusion. The increase rate of PRL induced by VIP was higher in male rats (158.04 +/- 37.06), but lower in the female (Diestrus: 50.42 +/- 16.44, Proestrus: 62.67 +/- 21.34) and in Suckling-depended lactating ones (Suckled 90.00 +/- 36.00 vs. Separated 31.05 +/- 4.42). The above observations suggest that the VIP action in vivo depends on the endocrinal and/or neural status of the animals. PMID- 11499003 TI - [Destruction of NA innervation in the rat hippocampus resists the hypoxia-induced corticosterone secretion]. AB - With microinjection of 6-hydroxydopamine (6-OHDA) into the ventral hippocampus (VHIP) of rats, the effect of 6-OHDA lesion of hippocampal noradrenergic system on the plasma corticosterone (CS) response to acute hypoxia was investigated. It was found that the plasma CS response to inhalation of 10.4% O2 (30 min) was increased markedly; while the plasma CS level and the noradrenaline (NA) contents in hippocampus were decreased (-33.2% and -38.5%, respectively) 2 weeks after microinfusion of 6-OHDA (8 micrograms/2 microliters) into VHIP; however, the CS response to acute hypoxia was unaffected by 6-OHDA lesion of VHIP. The results above suggest that NA in hippocampus may participate in the regulation of the hypoxia-induced increase of plasma CS concentration in rats. PMID- 11499004 TI - [Glycine modulates the ERG b-waves and ON-type bipolar cells in carp retina]. AB - In the present work we investigated the effects of glycine on the electroretinograms (ERG) and light responses of ON-type bipolar cells under dark- and light-adapted conditions in superfused, isolated crucian carp retinas. It was revealed that application of 4 mmol/L glycine significantly suppressed the b waves and the responses of ON-type bipolar cells and the effects were blocked by co-application of 20 mumol/L strychnine. In addition, glycine had no apparent action on the PIII component (photoreceptor potential) isolated by application of 3 mmol/L glutamate. These results suggest that glycine may act on bipolar cells directly and thus modulate the activity of the ON pathway in the retina. PMID- 11499005 TI - [Protection of spinal motorneurons of section sciatic nerve by transplantation of NT-4 genetically expression cells into the side of section]. AB - The NT-4 genetically engineered cells were made by infecting L-6TG cell (a rat myoblast cell line) in vitro with a retroviral vector pN2A containing the rat NT 4 cDNA. The bioactivities were determined by bioassay of PC12 cell survival rate. The rat with left sciatic nerve transaction was used as a model for treatment by implanting NT-4 genetically modified cell. The condition of motorneurons was assessed by Nissl stain and ChE stain. The results showed that: (1) the percentage of surviving Nissl-stained neurons on the lessened side of NT-4 (+) grafts significantly increased as compared to that with NT-4 (-) grafts 2-3 weeks and 3 months after sciatic nerve transaction, and (2) the grafted cells produced significant increase in the positive ChE stained area after sciatic nerve transaction in 1-3 weeks. Our observations indicate that adult motor neurons are still able to respond to neurotrophic factors and they may require the factors for survival. PMID- 11499006 TI - [Evidence for involvement of NO/NOS-cGMP signal system in morphine dependence]. AB - The present study was undertaken to observe changes in cGMP contents, calcium dependent and non-calcium-dependent NOS activities in brain regions isolated from morphine-dependent mice as well as the effect of NOS inhibitor (L-NMMA) on the development of this dependence. It was found that (1) cGMP contents in cerebellum, striatum, hippocampus and cerebral cortex were significantly decreased. (2) Calcium-dependent NOS activity was noticeably increased in striatum and cerebral cortex, which was inhibited by PKA inhibitor. No similar changes were found in cerebellum and hippocampus. Changes of non-calcium dependent NOS activity did not occur in morphine-dependent mice brain. (3) In the striatum and cerebral cortex of morphine-dependent mice, the level of 150 kD protein phosphorylation in vitro was noticeably decreased, which was inhibited by IP20 (PKA inhibitor). (4) NOS inhibitor injected (icv) 15 min prior to daily morphine injection could prevent the development of morphine dependence. (5) All the changes above were not observed in mice treated with naloxone 30 min prior to daily morphine injection. Our data suggest that the reduction of cGMP contents and the increase of calcium-dependent NOS activity in striatum and cerebral cortex isolated from morphine-dependent mice may be mediated by opioid receptors and involved in the development of morphine-dependence. Why the increase of NOS activity was in association with the reduction of cGMP contents remains to be answered and it implies that the effect of NO/NOS involved in morphine-dependence may be produced through other mechanisms other than those producing cGMP signal. NOS phosphorylation in some other brain regions, which may be regulated by PKA, probably contributes to the increase of NOS activity in morphine-dependent mice. PMID- 11499007 TI - [Effect of intracerebroventricular neurotensin injection on blood pressure in rat]. AB - Injection of neurotensin (NT) (10 and 20 micrograms/rat) into the lateral cerebroventricle (icv) could induce a rise or a drop of blood pressure and a slowing down of heart rate in urethane anaesthetized male rats (1.2 g/kg). The central pressor and depressor response to NT could be respectively blocked by pretreatment with alpha 1-receptor blocker prazosin (1.5 micrograms/3 microliters, icv) and M receptor blocker atropine (2.5 micrograms/3 microliters, icv). No significant changes in central cardiovascular responses to NT were observed following pretreatment with either H1 receptor blocker chlorpheniramin (50 micrograms/3 microliters, icv) or H2 receptor blocker cimetidine (250 micrograms/3 microliters, icv). In view of the above results it appears that NT, catecholamine and acetylcholine are all involved in central regulation of blood pressure. PMID- 11499008 TI - [Rapid effects of hydrocortisone on the cholinergic synaptic transmission of B neurons in bullfrog sympathetic ganglia]. AB - Rapid effects of hydrocortisone 21-hemisuccinate (F-suc) on cholinergic synaptic transmission of B neurons in bullfrog sympathetic ganglia (BFSG) were studied with intracellular recording technique in vitro. The main results are as follows: (1) F-suc blocked cholinergic synaptic transmission of 52 B neurons within 0.5-3 min, which was concentration-dependent and partially suppressed by the cytoplasm receptor antagonist of steroid hormones, RU38486; (2) after protein synthesis was suppressed by actidione, the above effect persisted; and (3) the effect of F-suc could be potentiated by atropine. All these results suggest that the F-suc induced rapid effects of cholinergic synaptic transmission of B neurons in BFSG may be mediated by non-genomic mechanisms. PMID- 11499009 TI - [Potentiation of caffeine-induced contracture by raising extracellular potassium in frog skeletal muscle]. AB - The effect of raising extracellular potassium ([K+]o) on caffeine contracture was investigated, using small bundles dissected from frog anterior tibialis muscle. Elevating [K+]o from the control of 2 mmol/L to 10 or 25 mmol/L significantly potentiated the contracture induced by 3 mmol/L caffeine. The potentiation represented by PKC/PC, where PKC and PC are the peak tension of the caffeine contracture evoked in high and normal [K+]o respectively, was dependent on [K+]o and the duration of conditioning high K+ exposure. With 10 mmol/L [K+]o, the potentiation was gradually increased by prolonging conditioning exposure up to 10 min. On the contrary, with 25 mmol/L [K+]o the potentiation reached a maximum within only 1 min, and then subsided to the control. These different time courses of PKC/PC could not be accounted for by high K+ induced depolarization, but were in general consistence with the time courses of the change in myoplasmic free calcium induced by corresponding high [K+]o. It is suggested that, at least in frog skeletal muscle, the high [K+]o induced potentiation of caffeine contracture is mainly due to an increase of myoplasmic free calcium. PMID- 11499010 TI - [Antipyretic action of dexamethasone on egtazic acid-induced fever in rabbits]. AB - The purpose of the present study was to investigate whether the antipyretic effect of dexamethasone (DEX) delivered by intravenous injection (i.v.) on intracerebroventricularly (icv) administered egtazic acid-induced febrile response is relevant to the changes in cytosolic free calcium concentration of the hypothalamus. The colon temperatures were measured by a thermistor and the cytosolic free calcium concentration ([Ca2+]i) in dissociated brain cells was measured by Fura 2-AM. The results demonstrated that the pyretic action of egtazic acid (0.6 mumol, icv) was markedly inhibited by DEX (5 mg/kg, i.v.), but DEX (60-120 mumol/L) did't affect [Ca2+]i in dissociated hypothalamus cells. Actinomycin D, which interferes with gene transcription (3 nmol, icv), completely abolished the antipyretic action of DEX on egtazic acid-induced fever. These findings suggest that the antipyretic action of DEX on egtazic acid-induced fever is related to the activation of certain gene expression in the brain, but not to the changes of transmembrane calcium ion current in hypothalamus neurons. PMID- 11499011 TI - [The coherence analysis between neuronal discharge in the rostral ventrolateral medulla and the cardiovascular activity in rats]. AB - To analyze the coherence between neuronal discharges (ND) in the rostral ventrolateral medulla (RVLM) and the cardiovascular activity, we observed the neuronal discharge in RVLM responding to electric stimulation of the defense area of the mid-brain. Fast Fourier transform (FFT) was performed to analyze the coherence between the signals of ND and blood pressure to determine if the ND were cardiac rhythmic. The coherence between ND variability (NDV) and heart rate variability (HRV) was also analyzed. The results showed: (1) majority of the neurons (67%) were excited responding to electric stimulation in the defense area of the mid-brain; (2) the electric activity of about 70% of the neurons were substantially inhibited by administration of phenylephrine; (3) 64% of the neurons were actively synchronous with cardiac cycle; and (4) significant coherence between NDV and HRV in HF component was shown in a half of the neurons (50%). The coherence analysis thus provides a new tool to investigate the regulation of the autonomic nervous system. PMID- 11499012 TI - [Inhibitory effect of electroacupuncture on the cardiovascular response evoked by applying bradykinin on the gallbladder]. AB - The effects of electroacupuncture (EA) on the pressor response and reversible myocardial dysfunction induced by application of bradykinin (BK) on the gallbladder were studied in cats anesthetized with alpha-chloralose. The cardiovascular responses evoked by application of BK included a pressor response, an increase of LVP and its dP/dtmax, tachycardia and a decrease of local wall motion of the left ventricle with a supplying branch of the left anterior descending coronary artery ligated beforehand. Following EA of bilateral Neiguan acupionts, the pressor response of BK was inhibited, while the regional left ventricle myocardial dysfunction was alleviated significantly. The effects of EA were reversed by i.v. injection of naloxone (0.4 mg/kg). Our results indicate that EA has an inhibitory effect on the BK-induced pressor and ischemic dysfunction, which may be related with endogenous opioid peptide. PMID- 11499013 TI - [Macrophage colony-stimulating factor production and c-fms expression in cultured rat aortic vascular smooth muscle cells]. AB - The resulats of this study are as follows. (1) As measured by a bioassay, a macrophage colony-stimulating activity was detected in the serum-free conditioned medium of rat aortic vascular smooth muscle cells (VSMCs), which could be subdued by the addition of specific anti macrophage colony-stimulating factor (MCSF) antibody. (2) The presence of MCSF receptor was confirmed by immunocytochemistry using a specific anti c-Fms antibody. (3) The presence of mRNAs for MCSF and c fms (which encoded MCSF receptor) was determined by Northern blot analysis. Their expressions were detectable in quiescent VSMCs and markedly increased after addition of serum. These data demonstrated for the first time the production of MCSF and the presence of MCSF receptor in cultured rat VSMCs. It is suggested that MCSF might modulate VSMCs functions via both autocrine and paracrine mechanisms. Rat VSMCs appear to be a suitable cell model for studying the cell proliferation effect of MCSF. PMID- 11499014 TI - [Effects of intermittent hypoxia on transient outward current in rat ventricular myocytes]. AB - To explore the ionic basis of the strengthening effect of intermittent hypoxic adaptation (IHA) on the electric stability of heart, the effects of intermittent hypoxia on the transient outward current (Ito) in rat ventricular myocytes were investigated by using whole-cell patch-clamp recording techniques. After 28-day (H28) exposure (6 h/d) to intermittent hypoxia, the density of Ito in the right, but not in the left, ventricular myocytes was dramatically increased as compared with the normoxia control (16.18 +/- 4.61 vs 6.32 +/- 1.35 pA/pF, P < 0.05), while the Ito density of the myocytes isolated from both sides of ventricles in 42-day-exposure group (H42) did not show significant difference. Except for a more negative shift of the steady-state inactivation curves (half-inactivation voltages: -38.9 +/- 2.3 vs -32.8 +/- 5.9 mV in the left ventricle and -41.9 +/- 4.5 vs -33.5 +/- 3.5 mV in the right ventricle) in the H42 group, all the other parameters for activation, inactivation and recovery kinetics of Ito of each group remained unchanged. It is speculated that the change in the current density of Ito may be responsible for the different hemodynamic responses of the ventricles to the early stage of hypoxia. The alteration in inactivation may participate in the cardioprotective effect of IHA. PMID- 11499015 TI - [Expression of c-fos in the medulla oblongata after carotid baroreceptor activation by elevated intrasinus pressure and adenosine]. AB - Expression of c-fos protein in the medulla oblongata after baroreceptor activation by elevated intrasinus pressure (ISP) and perfusion of adenosine (Ado) was examined in 14 vascularly isolated carotid sinus perfusion rats. The results showed that Fos-like immunoreactive (FLI) neurons were distributed throughout nucleus tractus solitarius, area postrema, rostral ventrolateral medulla and nucleus raphe pallidus, and the number of FLI was increased with the elevation of ISP. Furthermore, perfusing the carotid sinus with Ado at a given ISP markedly increased the FLI in the above regions. From the results obtained, it is concluded that the c-fos expression in baroreflex pathway in medulla oblongata may be enhanced by elevated ISP and intrasinus perfusion of Ado, and Ado is capable of facilitating the baroreflex. PMID- 11499016 TI - [Expression of human superoxide dismutase gene in rat vascular smooth muscle cells and its anti-oxidative effect]. AB - A retroviral vector containing human superoxide dismutase (SOD) cDNA was constructed and transfected into rat vascular smooth muscle cells (VSMCs). The expression of exogenous hSOD1 in the VSMCs was analyzed with Northern and Southern blot. The protection of the transfected and/or non-transfected VSMCs from free radical produced by the xanthine/xanthine oxidase (X/XO) system was investigated. The results showed that the construction strategy of the vector was correctly performed and the expression of hSOD1 in the transfected cells was highly detectable. The cell damage of X/XO could be alleviated with expression of hSOD1 in the transfected cells, as compared to control. In addition, proliferation of the transfected VSMCs resulted from oxidative stress was suppressed. It is suggested that the expression of gene-transferred hSOD1 is able to prevent the formation of atherosclerosis, partially due to its cell protection and inhibition of the proliferative embolization. PMID- 11499017 TI - [The role of NO/ET and the effect of electroacupuncture on injuried gastric mucosa in rats]. AB - The aim of the present work is to observe changes in endothelium-derived factors- nitric oxide (NO) and endothelin (ET), with special reference to the regulative role of electroacupuncture (EA) in injured gastric mucosa of rat by ethanol. It was found that gastric mucosal blood flow (GMBF), transmucosal potential difference (PD) and the content of NO in the serum all decreased (P < 0.01), while the content of ET in the plasma and the lesion index (LI) of gastric mucosa increased (P < 0.01). After administration of L-Arg and SNP, the content of NO and GMBF increased significantly (P < 0.01), whereas the content of ET and LI decreased (P < 0.01). These effects were enhanced by EA at ZuSanLi (ST-36) points, which, however, were inhibited by NO biosynthetic inhibitor--L-NNA. The latter effect could be reversed by concurrent administration of L-Arg. The above results suggest that NO plays a critical part in protecting gastric mucosa against injury, which is also responsible for the protective effect of EA on injury of gastric mucosa. PMID- 11499018 TI - [Modulation of the secretion of active renin in human decidual cells by progesterone]. AB - Uterine decidua is a major source of renin, both active and inactive. Active renin cleaves angiotensinogen to produce angiotensin I and exerts its effect on the expression of angiotensin II in the local tissue. Our experiments showed that (1) in the decidua of early pregnancy the contents of active renin increased gradually with advancing pregnancy, and at 8th week of gestation the level of active renin reached to a maximum (63.37 +/- 12.84) AI ng/gww.h(-1); (2) active renin accounted for about 1/4 of the total renin; and (3) the synthesis and secretion of active renin were regulated by progesterone in incubated decidual cells. These data indicate that a high level of active renin in decidual tissue at early pregnancy plays an important role in the uterine renin-angiotensin system during pregnancy. PMID- 11499020 TI - [Effect of anti-opioid peptide sera on the enhancement of electroacupuncture analgesia induced by neurotensin in PAG of rats]. AB - With the use of potassium iontophoresis induced tail-flick for measuring the pain threshold, the effects of injecting neurotensin (NT), naloxone (NX), anti metenkephalin serum (AMEKS), anti-beta-endorphin serum (AEPS) and anti-dynorphin A1-13 serum (ADYNS) into periaqueductal gray (PAG) on electroacupuncture (EA) analgesia in rats were investigated. NT administration enhanced EA analgesia remarkably. Pre-injection of NX, AMEKS and AEPS into PAG could significantly attenuate the enhancement of EA analgesia induced by NT, but not by administration of ADYNS. The results indicate that NT in PAG is responsible for the enhancement of EA analgesia. The effect of NT may be partly mediated by met enkephalin and beta-endorphin. PMID- 11499019 TI - [The excitatory effects of histamine on cerebellar cortical Purkinje cells in the rat]. AB - The effects of histamine on Purkinje cells (PCs) in lobule X of cerebellar cortex were investigated in rat cerebellar slices. Histamine elicited PCs' responses predominately excitatory (94.4%, 51/54), only few inhibitory (5.6%, 3/54). The excitatory response could not be abolished by perfusing the slices with low Ca2+/high Mg2+ medium. The excitatory response of PCs to histamine could be blocked by histamine H2 receptor antagonist ranitidine, but not by H1 receptor antagonist triprolidine readily. These results suggest that histamine exerts an excitatory effect on the PCs via H2 receptors. Presumably, hypothalamo-cerebellar histaminergic fibers participate in the regulation of somatic and non-somatic functions of the cerebellum. PMID- 11499021 TI - [Hemodynamic effects of agmatine and its cellular mechanism in anesthetized rats]. AB - The hemodynamic effects of intravenous injection of agmatine and their cellular mechanism were investigated in anesthetized rats. The results obtained are as follows. (1) Following intravenous injection of agmatine (10 mg/kg), HR, MAP, LVP, +/- LV dp/dtmax, CI and TPRI were significantly decreased. (2) Pretreatment with N-nitro-L-arginine (15 mg/kg) or methylene blue (50 mg/kg), did not affect the hypotensive effect of agmatine. (3) The hemodynamic effects induced by agmatine could be inhibited by prior intravenous injection of idazoxan (2 mg/kg), an alpha 2-adrenoceptor (alpha 2-AR) and imidazoline receptor antagonist. The results indicate that the hypotensive effect induced by i.v. agmatine may be attributed to the decrease in cardiac output resulting from depression of myocardial contractility, as well as to the reduction in total peripheral resistance resulting from vasodilatation. These effects of agmatine may be mediated by imidazoline receptor and/or alpha 2-AR. PMID- 11499022 TI - [Establishment of an apoptosis model of vascular smooth muscle cells induced by ultraviolet-C radiation in vitro]. AB - After a 10-min exposure to 254 nm UV-C radiation, aortic SMCs of rat were found to undergo apoptosis, characterized by cell shrinkage, condensation of cytoplasm, membrane blebbing, appearance of apoptotic body and DNA ladder in the agarose gel. Therefore, UV-C radiation can be used to generate a useful model for the investigation of the molecular regulatory mechanisms underlying SMCs apoptosis. PMID- 11499023 TI - [Differential regulation of dopamine receptors on pre- and postsynaptic Na+, K(+) ATPase in rat striatum]. AB - The pre- and postsynaptic membranes isolated from rat striatum were used to investigate the regulation of dopamine receptors on striatal Na+, K(+)-ATPase in these membranes. The activity of Na+, K(+)-ATPase was determined by colorimetric method. Dopamine (DA) was found to inhibit the Na+, K(+)-ATPase activity on postsynaptic membranes in a concentration-dependent manner with a IC50 value of 4.6 mumol.L-1. This inhibitory effect was reversed by either selective D1 receptor antagonist SCH23390 or selective D2 receptor antagonist spipernone. The inhibitory effect similar to DA was produced by combination with selective D1 receptor agonist SKF38393 and selective D2 receptor agonist LY171555. In contrast, under the same experimental conditions, DA (10(-8)-10(-5) mol.L-1) was shown to activate the activity of Na+, K(+)-ATPase on presynaptic membranes in a concentration-dependent manner. Meanwhile, the stimulatory effect was reversed by spiperone alone rather than by SCH23390. These results show the differential regulation of presytnaptic and postsynaptic DA receptors on Na+, K(+)-ATPase in rat striatum. PMID- 11499024 TI - [A comparative study on effect of two bisbenzylisoquinolines, tetrandrine and berbamine, on reversal of multidrug resistance]. AB - A comparative study on the effect of two bisbenzylisoquinolines, tetrandrine (TTD) and berbamine (BBM), and verapamil (VRP) on reversing multidrug resistance was reported. TTD, BBM and VRP showed significant activity in reversing adriamycin (ADR) and vincristine (VCR) resistance in acquired resistant MCF-7/Adr and KBv200 cell lines, and the effect was shown to be dose-dependent. TTD, at the concentration of 10 mumol.L-1, completely reversed ADR resistance in MCF-7/adr cells. TTD, BBM and VRP increased intracellular ADR accumulation in MCF-7/adr cells. There is minor difference in structure between TTD and BBM. TTD showed greater activity than VRP in reversing MDR, while BBM showed similar activity to that of VRP. TTD also showed significant activity in vivo in reversing ADR resistance in MDR MCF-7/Adr solid tumor in nude mice. PMID- 11499025 TI - [Effect of age and ginsenoside Rg1 on nitric oxide content and nitric oxide synthase activity of cerebral cortex in rats]. AB - Nitric oxide(NO) content and nitric oxide synthase(NOS) activity were measured in cerebral cortex isolated from Wistar rats of three age groups(young: 3 months; adult: 9 months; and old: 27 months). No significant differences in NO content and NOS activity between young and adult rats were found(P > 0.05). The NO content and NOS activity in old rats were shown to be significantly higher than those of young and adult rats(P < 0.01). When treated with Rg1(10, 20, 40 mg.kg 1), the NO content and NOS activity in old rats decreased. The inhibitory effect of Rg1 on NOS was found to be dose-dependent in the range of 10-40 mg.kg-1. The optimal reduction in NO content and NOS activity induced by Rg1 occurred at 40 mg.kg-1 for old rats(P < 0.01). In view of the close relationship of NO content and NOS activity with aging, the inhibitory effect of Rg1 on NOS activity, as shown by our results, might provide an explanation for its antiaging function. PMID- 11499026 TI - [Rolipram reversed salbutamol tolerance in guinea pig trachea]. AB - Studies were carried out to examine the role of phosphodiesterase(PDE) III and PDE IV in the development of tachyphylaxis of isolated guinea pig trachea to beta 2 adrenoceptor agonists. Treating trachea with salbutamol(SB) 1 mumol.L-1 for 1 h produced a 5-fold rightward shift of the SB concentration-response curve for the spasmolytic effect against methacholine-induced bronchocontraction and decreased the maximum SB-induced relaxation by 30%, i.e. induced tolerance of airway response of SB in vitro. The PDE IV inhibitor rolipram (Rol, 1 mumol.L-1, IC50), but not the PDE III inhibitor siguazodan (SK&F 94836, 1 mumol.L-1, IC50), reversed the SB tolerance. However, the protein synthesis inhibitor cycloheximide(10 mumol.L-1) did not abolish the SB tolerance. These results indicate that the SB tolerance may be related to increase in PDE IV activity. PMID- 11499027 TI - [Effects of (-), (+) clausenamide on central N-methyl-D-asparate receptors in rodents]. AB - Using radioligand binding assay, the effects of (-), (+) clausenamide on N-methyl D-asparate(NMDA) receptor were studied in synaptic membrane, hippocampus and cerebral cortex in rats. The Bmax and KD values of NMDA receptor in mouse brain were measured with Scatchard plot method. Results showed that there was no specific binding of (-), (+) clausenamide to NMDA-receptor. However, higher Bmax values were observed in (-) clausenamide-treated rats than the control group, but no effect on KD value. (+) Clausenamide treatment showed no effect on Bmax and KD values. The findings suggest that the pharmacologic actions of clausenamide depends on its chirality. Up regulation of NMDA-receptor induced by (-) clausenamide is helpful to elucidate its nootropic mechanism. PMID- 11499028 TI - [Synthesis and antiarrhythmic activity of some (erythro)-phenylpropanediolamine compounds]. AB - For the purpose of searching for new drug with high potency and simple chemical structure, the dominant conformation and structural parameters of Guan-Fu base (GFA) molecule were modelled and calculated with a SGI-4D 25G computer. The propanediolamine chain in GFA might be considered to be a pharmacophore responsible for the bioactivity and the configuration of the chain seemed important. Thus, thirteen compounds of (erythro)-p-x-PhCHOHCHOHCH2NHR(x = H, I1 7; X = NO2, II1-6) were prepared. Among them, 10 compounds showed antiarrhythmic effect on aconitine-induced arrhythmia in rats. The ED50(to stop VT) of I2 and ED50(to stop VP) of I3 were shown to be comparable with those of GFA. In the synthesis, no stereoselectivity was found in the Prevost reaction with allylamine analogues (a1-7). After a1-7 were acetylated, the erythro type products(I1-7) were obtained. PMID- 11499029 TI - [Studies on the synthesis and activities of RGD related peptides]. AB - In the binding of Fgn to GP IIb/IIIa, RGD is the key sequence. In the present paper, RGDS, RGDV and RGDF were synthesized by use of solution method. Bioassay indicated that the C-terminal amino acid residues were very important for their antithrombosis effects. Conformational studies showed that their antithrombosis potency may depend on their total energies. The observation of their vasodilation effects suggests that this kind of function is worthy to be further studied. PMID- 11499030 TI - [Total synthesis of nordihydroguaiaretic acid]. AB - beta-Keto ester(5) was obtained from vanilin through etherification, oxidation and condensation with acetoacetic ester, (5) on oxidative coupling reaction by NaOEt/I2 produced dimer (6) in high yield. Acid catalyzed cyclodehydration of (6) gave the furan derivative(7), and by a series of selective hydrogenation nordihydroguaiaretic acid, furoguaiacin dimethyl ether and dihydroguaiaretic acid dimethyl ether were synthesized. PMID- 11499031 TI - [Studies on liposoluble constituents from the aerial parts of Siegesbeckia orientalis L]. AB - Eight compounds were isolated from the aerial parts of Siegesbeckia orientalis L. and their structures were elucidated by spectroscopic methods(IR, EI-MS, 13C-NMR, 1H-NMR, 1H-1H COSY, 1H-1H NOESY and 1H-13C COSY). Compounds I and II are new natural products and named siegesesteric acid(I) and siegesetheric acid(II), their structures were confirmed as ent-17-acetoxy-18-isobutyryloxy-16(alpha) kauran-19-oic acid(I) and ent-17-ethoxy-16(alpha)-(-)-kauran-19-oic acid(II). The others were identified as known compounds: ent-16 beta, 17-dihydroxy-kauran-19 oic acid (III), kirenol(IV), beta-sitosteryl glucoside(V), heneicosanol(VI), methyl arachidate(VII) and beta-sitosterol(VIII). These compounds, except kirenol and beta-sitosterol, were isolated for the first time from the title plant. PMID- 11499032 TI - [Studies on new cytotoxic annonaceous acetogenins from Uvaria grandiflora and absolute configurations]. AB - Two new cytotoxic annonaceous acetogenins, named uvarigrin(1) and uvarigrandin A(3), were obtained from the roots of Uvaria grandiflora Roxb(Annonaceae). Based on X-ray analysis and Mosher's methodology, the overall absolute configuration of 1 was established as 15S, 17R, 18R, 21R, 22R, 36S. The absolute configuration of 3 was also resolved by Mosher's methodology. The relative configuration of the previously reported uvarigranin (2) was revised. Compound 1 showed cytotoxicity against HCT-8, Bel7402 and A2780 human tumor cell lines at ED50 levels of 0.15, 0.21 and 0.41 microgram.ml-1, respectively. PMID- 11499033 TI - [Analysis of phenylethanoid glycosides of Herba cistanchis by RP-HPLC]. AB - The Chinese drug "Rou Cong-rong" (Herba Cistanchis) is one of the commonly used drugs in Chinese traditional medicine. It is used to reinforce the vital function of kidney, especially that of the sexual organs and induce laxation, for the treatment of impotence, premature ejaculation in men, infertility, morbid leukorrhea, profuse metrorrhagia in women, and chronic constipation in the aged. This paper deals with the qualitative and quantitative analysis of phenylethanoid glycosides of four species and one variety of Genus Cistanche and 23 lots of commercial crude drugs of Herba Cistanchis by RP-HPLC. The results were as follows: the chemical constituents of Cistanche deserticola Ma, C. salsa (C. A. Mey) G. Beck, C. salsa var. albiflora P. F. Tu et Z. C. Lou and C. tubulosa were similar while those of C. sinensis were different from the others; the contents of echinacoside and acteoside of C. salsa, which were 2.13% and 1.51%, were the highest of the genus Cistanche. An ODS column (Alltima C18, 5 microns, 250 x 4.6 mm) was employed. Linear gradient elution of acetonitrile--1.5% acetic acid was used as mobile phase, and concentration of acetontrile was from 8% to 20% (0-60 min) in the qualitative analysis, and from 11.5 to 20% (0-35 min) in the quantitative analysis. The flow rate was 1.2 ml.min-1. The detection wavelength was set at 335 nm. PMID- 11499034 TI - [Isoflavones from Glycyrrhiza eurycarpa]. AB - A new isoflavone, named eurycarpin A and a new natural product isoflavone named eurycarpin B have been isolated from the roots of Glycyrrhiza eurycarpa P. C. Li. Their structures were determined to be 7,2',4'-trihydroxy-3'-(3,3-dimethylallyl) isoflavone(I) and 7,2'-dihydroxy-6",6"-dimethylpyrano-(2",3":4',3') isoflavone(II) on the basis of spectroscopic analysis (UV, EI-MS, 1HNMR, 13CNMR, NOE difference and HMBC). In addition, three known isoflavones, licoisoflavone A, calycosin and formononetin, were obtained for the first time from this plant. PMID- 11499035 TI - [Studies on the analysis of hydrocodone and its metabolite in human urine by GC/MS]. AB - The method for the analysis of hydrocodone and its metabolite in urine by GC/MSD is reported. The urine was hydrolysed with HCl and extracted with diethyl ether/isopropanol. The gas chromatography/mass spectrometric properties of trimethylsilyl ether of hydrocodone, first reacted with methoxyamine to protect the carbonyl group, were studied. This method is sensitive and rapid for the determination of hydrocodone, codeine and dihydrocodeine. PMID- 11499036 TI - [Investigation on electrochemical behavior of emodin and its application]. AB - Emodin showed a second-order derivative reduction wave with peak potential of 0.75 V (SCE) by single sweep oscillopolarography using H3BO3-Na2B4O7 (pH 8.50) as base solution. The peak height of emodin is proportional to the concentration in the range of 1.42 x 10(-7)-5.7 x 10(-6) mol.L-1 and 7.1 x 10(-6)-7.1 x 10(-5) mol.L-1, respectively. The detection limit is 0.7 x 10(-7) mol.L-1. It can be employed for the determination of emodin in Rheum officinale Baill with satisfaction. The electrochemical behavior of emodin was studied and the electrode reaction mechanism was proposed. In addition, it was found that emodin, aloe-emodin, chrysophanol, rhein and physcion can scavenge superoxide anion radical produced by the autoxidation of pyrogallol. The scavenging activity is in the order: emodin > aloe-emodin > chrysophanol > rhein > physcion. PMID- 11499037 TI - [Interactions between ciprofloxacin, Mg2+, Mn2+ and DNA by electrochemical method]. AB - Interactions between ciprofloxacin (CPFX), Mg2+, Mn2+ and DNA, and their polarographic and voltammetric behaviour were studied. In 0.1 mol.L-1 NH3-NH4Cl buffer solution (pH 9.2), a new reduction peak was obtained by linear-sweep voltammetry with Ep = -1.72 V(vs Ag/AgCl) when adding DNA to CPFX solution, which implies binding of CPFX with DNA. In the presence of Mg2+ or Mn2+, another new sensitive reduction peak, whose peak potential is more negative (Ep = -1.78 V), was obtained which suggested that Mg2+ or Mn2+ took part in the interaction between CPFX and DNA resulting in a ternary complex of CPFX-Mg-DNA. The peak current (ip) is proportional to the concentration of DNA over the range of 1.18 x 10(-4)-3.33 x 10(-4) mol.L-1. In this paper, the properties of the peak current were studied in detail, the result showed that the electrode reaction was irreversible and the ip was influenced by adsorption. The electrode reaction mechanism was also probed into. The CPFX molecule in the complex was reduced on the electrode. Furthermore, CPFX-Mg was shown ot be intercalated between the stacked base pairs of native DNA. PMID- 11499038 TI - The ePhilanthropy revolution. AB - "The e-philanthropy revolution is here to stay, and it will transform charitable giving in as profound a way as technology is changing the commercial world. Charities that have dismissed e-philanthropy, as a fad, or run from it in confusion, will, sooner or later, need to become reconciled to it. If they don't, they risk losing touch with donors and imperiling the vitality of their work." PMID- 11499039 TI - Cutting through the e-maze: marketing strategy (Part I). AB - If we, as development officers, are doing our job properly, we should be getting to know our donors in the early stages of their giving pattern when he/she is in good health and their giving potential is maximized. During these years, emphasis should be placed on donor acquisition and retention. PMID- 11499040 TI - Get to know your allies. AB - With time at a premium, wealthy people rely on a variety of professional advisors to help them make good decisions about many of their financial choices, and those can include decisions about charitable donations. PMID- 11499041 TI - XXIII John Caples international non-profit award winners. AB - The following non-profit direct marketing campaigns took honors at this year's John Caples International Awards, held at the New York Marriott Marquis Hotel, New York, NY. PMID- 11499042 TI - Ask ... the secret to fund raising. AB - If we, as development officers, are doing our job properly, we should be getting to know our donors in the early stages of their giving pattern when he/she is in good health and their giving potential is maximized. During these years, emphasis should be placed on donor acquisition and retention. PMID- 11499043 TI - Listen and learn. AB - After making a solicitation, a lot can be gained by listening to a donor's questions. Do not simply answer the question. Consider why they asked it in the first place. PMID- 11499044 TI - How to use post cards to capture readers--and keep them begging for more. PMID- 11499045 TI - Clinical guidelines: attitudes, information processes and culture in English primary care. AB - The application to clinical medicine of evidence-based clinical guidelines is an increasingly international policy prescription, yet research on how such guidelines might be implemented has tended to focus on change initiatives without seeking to understand change processes. This paper reports an empirical study of guideline implementation in UK general practice. Most GPs welcome guidelines as a means of improving care, though have reservations about their authority, relevance and effect on professional autonomy. 'Clan' organizational culture predominates and general practices do not generally have well-functioning internal arrangements for the management of clinical evidence and related information. We found no coherent relationships between these variables and practices' actual uptake of guidelines. PMID- 11499046 TI - The practice characterization model: the importance of organizational life cycles and targeted interventions in general medical practice. AB - In response to a climate of constant change and increasing demand for services, general practice in the UK has undergone significant modification over the last 10 years. It has become a multi-disciplinary organisation encouraged by funding bodies to plan for service delivery using a more structured team based approach. In Tayside in 1996, practices were charged with producing formal Practice Development Plans (PDPs) which would focus on priority areas aligned with the Health Boards own strategic plan--those were teamwork, information management and technology, and clinical service delivery. The University of Dundee's Department of General Practice successfully applied for funding to develop ways of facilitating practices so that they could a) identify their own development priorities, and b) plan and implement action and learning to see these priorities through. Using action research methodology, the project attempted to create a climate for change, provide support and training to see the changes implemented, and ensure commitment to the changes from all members of the practice team. The Facilitator adopted a flexible style varying her role between expert, guide and support. Analysis of progress made by different practices, coupled with the Facilitator's in depth knowledge of them, suggested the importance of certain key aspects of practice organisation and culture. A practice characterisation model identified practices which were stable, currently coping, proactive and ready to face the challenge of change as best placed to engage in a full scale development programme. Other profiles suggested a range of alternative interventions as more likely to be acceptable and productive. PMID- 11499047 TI - On a hiding to nothing? Assessing the corporate governance of hospital and health services in New Zealand 1993-1998. AB - In New Zealand the governance of public sector hospital and health services has changed significantly over the past decade. For most of the century hospitals had been funded by central government grants but run by locally elected boards. In 1989 a reforming Labour government restructured health services along managerialist lines, including changing governance structures so that some area health board members were government appointments, with the balance elected by the community. More market oriented reform under a new National government abolished this arrangement and introduced (1993) a corporate approach to the management of hospitals and related services. The hospitals were established as limited liability companies under the Companies Act. This was an explicitly corporate model and, although there was some modification of arrangements following the election of a more politically moderate centre-right coalition government in 1996, the corporate model was largely retained. Although significant changes occurred again after the election of a Labour government in 1999, the corporate governance experience in New Zealand health services is one from which lessons can, nevertheless, be learnt. This paper examines aspects of the performance and process of corporate governance arrangements for public sector health services in New Zealand, 1993-1998. PMID- 11499048 TI - Decentralization and human resource management in the health sector: a case study (1996-1998) from Nampula province, Mozambique. AB - Despite political, cultural and geographical diversity, health care reforms implemented in many developing countries share a number of common features regarding management and structural issues. Decentralization of decision-making from the central authority to local and provincial levels is generally regarded in the literature to be an important way of achieving a more equitable distribution of health care and better management practices, aligned with local priorities and needs. However, in the absence of clear guidelines, continuous monitoring and an adequate supply of financial and human resources, decentralization processes are more likely to have a low impact on the process of health care reform and can, to a certain extent, provoke inequalities between regions in the same country. This qualitative study in Nampula province, Mozambique, was conducted to assess the impact of decentralization, through an analysis of the viewpoints of provincial health managers regarding their perceptions of the process, particularly with regard to the management of basic and elementary nurses. Secondary data from Nampula provincial reports and documents from the Mozambican Health Ministry were also reviewed and comparisons made with the experiences of other developing countries. PMID- 11499049 TI - Can a purchaser be a partner? Nursing education in the English universities. AB - Since the early 1990s, public sector management in England has been exhorted to follow the example of the private sector, and 'quasi-markets' have been established, for example in the health service. A quasi-market also exists between the NHS and higher education for the purchasing (or procurement) of nursing education. This paper uses policy documents such as the National Health Service Executive Circular (March 1999) on 'Good Contracting Guidelines' for Non Medical Education and Training, plus other relevant literature on the commodification of higher education, quasi-markets and contract theory to examine this market, and the confusion of two rhetorics, those of competition and partnership. Nursing occupies a marginal place in higher education in England, having only recently become part of it. The emphasis of the quasi-market on the output of a trained 'fit for purpose' labour force combines with professional attempts to create an academic discipline, in complex ways which are as yet underanalysed. PMID- 11499050 TI - Prozac. A bitter pill. PMID- 11499051 TI - Principles of screening. AB - Over the past several decades, the principles by which screening tests are performed have slowly been developed and refined. The key distinction for the clinician to understand is the difference between diagnostic tests, which give a definitive answer, and screening tests, which identify who among the low-risk population is at high risk. PMID- 11499052 TI - Screening for neural tube defects. AB - Neural tube defects are separated into two main categories: (1) abnormalities of the skull and brain (anencephaly, acrania, and encephalocele) and (2) malformations of the spine (meningomyelocele or spina bifida). The cause of neural tube defects is not always clear, and include chromosomal abnormalities, single gene mutations, maternal disease, or maternal exposure to teratogens. Mostly the disorder emerges as a multifactorial trait. Routine screening for neural tube defects was introduced in the United Kingdom in the mid-1970s and the United States in the mid-1980s. The use of screening has resulted in a marked decline in the frequency of neural tube defects diagnosed at birth. PMID- 11499053 TI - Second-trimester biochemical screening. AB - The past years have seen considerable progress in the area of biochemical screening. Increasing data have now clearly shown the advantages of multiple markers, particularly beta-hCG over AFP alone. There continues to be considerable controversy over the best mathematic algorithm and which markers are best (e.g., beta-HCG, uE3, and so forth). There seems to be a plateau of detection frequencies at about 65% to 70% with current methodologies. Further work needs to be done, however, including some new approaches, if there is to be substantial improvement of screening sensitivity. The combination of biochemical with biophysical parameters as discussed elsewhere in this issue represents the next level of sophistication in the attempt to identify the highest proportion of abnormalities with the fewest false-positives. PMID- 11499054 TI - Trisomy 21. Second-trimester ultrasound. AB - Not every aspect of sonographic examination reveals karyotypic abnormalities. Ultrasound examination of a fetus with trisomy 21 generally reveals normal amniotic fluid, normal placentation, and normal fetal growth. In addition, other chromosomal abnormalities have many of the same sonographic findings as Down syndrome, and many findings have a large overlap with phenotypically normal fetuses. The importance of second-trimester ultrasound screening for Down syndrome has remained great because of its ease of use and relative effectiveness. Trained sonographers can adjust the relative risk for trisomy 21 and alter the need for genetic amniocentesis. It is important that parents understand the limitations of a screening test and the risks and benefits of possible subsequent confirmatory testing. If a major structural abnormality is identified on ultrasound, karyotype determination should be considered. Nuchal thickness in the first or second trimester remains the most clinically useful marker for trisomy 21. The predictive value of all the markers depends on the population studied and can be modified by a host of biochemical markers and historical factors. If fetal karyotype analysis could be performed without sampling through the uterus, prenatal diagnosis could be offered to all pregnant women, and screening would be unnecessary. Despite its limitations, ultrasound will have an important role in prenatal diagnosis at least until isolating and testing fetal cells from maternal blood or other sources becomes practical and widely available. Whether used alone or in conjunction with additional biochemical or molecular serum markers, ultrasound is an important and powerful tool in prenatal genetic evaluation. PMID- 11499055 TI - First-trimester biochemical screening for Down syndrome. AB - Of all markers evaluated for first-trimester biochemical screening for Down syndrome (DS), PAPP-A and free beta-hCG emerged as the most predictive. The combined test uses these markers in conjunction with nuchal translucency measurements, and is estimated to achieve a DS detection rate of 80% to 85% at a 5% false-positive rate. The integrated test, combining first-trimester sonographic and biochemical markers with second-trimester markers, provides a single estimate of a patients DS risk, and may yield a DS detection rate of 94% at a 5% false-positive rate. The acceptability and feasibility of this test, however, remain to be proved. PMID- 11499056 TI - First-trimester ultrasound. AB - There has been an increase in the use of fetal ultrasound in the first trimester. This article reviews the published literature with emphasis on fetal nuchal translucency (NT). When measured according to well-defined guidelines, increased NT identifies about 80% of chromosomal anomalies for a false-positive rate of about 5%. Increased NT may indicate the presence of structural defects or genetic disorders, and it is relatively frequently seen in monochorionic pregnancies prone to develop twin-twin transfusion syndrome. PMID- 11499057 TI - Prenatal diagnosis using fetal cells and free fetal DNA in maternal blood. AB - The presence of fetal cells and free fetal DNA in maternal blood offers an exciting opportunity for the development of safe noninvasive forms of prenatal diagnosis. Research in this field has, however, also indicated that their levels in the maternal circulation are increased in certain pregnancy-related disorders, such as preeclampsia. Their closer examination may shed new light on the underlying etiology of this enigmatic disorder. PMID- 11499058 TI - Prenatal genetic screening in the Ashkenazi Jewish population. AB - The Ashkenazi Jewish community is a unique and ideal population in which to provide multiple disease screening because detection rates are high (> 95%) by testing a limited number of mutations. The residual risk that remains is very low. In addition, the lessons learned from carrier screening in this community indicate that only through genetic counseling and education can screening in the general population gain wide acceptance and provide maximum benefit. PMID- 11499059 TI - Cystic fibrosis. AB - Although mutation detection rates have not universally reached the 95% detection level recommended by the American Society of Human Genetics and are not likely to exceed 90% for many populations in the foreseeable future, CF carrier screening will probably be offered routinely in the near future. Although CF carrier information will be of benefit to some individuals and couples, the inability of conventional prenatal diagnosis to provide definitive diagnostic outcomes to some couples, specifically those couples in which only one partner has a detectable mutation, will make for considerable anguish and concern for some. Because genetic screening and counseling is meant to provide information and alleviate concerns and fears, the potential for CF screening to result in such a contradictory effect is of a continuing concern to those who provide obstetric and genetic services. In a National Institute of Health-sponsored workshop, Menutti and colleagues recommended that those populations to which carrier screening should be offered might include individuals and couples in high-risk groups (e.g., Ashkenazi Jews, central or northern Europeans, one partner with CF, and individuals with a family history of CF) who seek preconception counseling, infertility care, or prenatal care. The workshop participants concluded, however, that before screening can be offered systematically to these individuals or couples, practice guidelines, educational materials for providers and patients, informed-consent protocols, and laboratory standards for testing must be developed. Further advances in DNA and protein analytic capabilities, such as microchip analytic systems and protein truncation assays, may make CF screening and diagnosis more accurate and less likely to result in equivocal outcomes. In addition, it is hoped that continuing improvements in CF therapies will increase the life expectancy and improve the quality of life for individuals affected with CF. Expanding our current knowledge of genotype-phenotype correlations will not only allow us better to predict clinical outcomes but also may improve our treatments for individuals with CF because more targeted therapies may be developed for CF caused by specific mutations. Nonetheless, educational and counseling issues will, for the foreseeable future, remain of critical importance to ensure appropriate clinical care to low- and high-risk individuals. PMID- 11499060 TI - Identifying and managing hereditary risk of breast and ovarian cancer. AB - In the past, all women with a family history of breast or ovarian cancer were considered to be at increased risk of cancer themselves. The discovery of BRCA1 and BRCA2 demonstrated that susceptibility to breast and ovarian cancer can be inherited by women as a single-gene autosomal dominant disorder. For such women, evaluation of family history is an important screening tool to identify the possibility of hereditary cancer risk but only genetic testing can provide definitive, individualized risk assessment. Women who have inherited mutations in BRCA1 or BRCA2 now have several medical management options to address their increased risk of cancer. A well-educated community of health care providers and patients can use hereditary risk assessment, including genetic testing, to improve health care. PMID- 11499061 TI - To screen or not to screen for gestational diabetes mellitus. The clinical quagmire. AB - Although there continues to be a lack of agreement about the most appropriate way to screen for GDM, screening remains the standard of care in this country. Universal screening of all pregnant women maximizes sensitivity but has significant financial implications because of its increased costs. Additional studies are needed that apply cost-analysis to various screening protocols to identify cost-effective screening strategies. PMID- 11499062 TI - Genetic implications for newborn screening for phenylketonuria. AB - The genetic implications for PKU are similar to those for any inherited disorder, but require an intimate knowledge of the dietary care required by these women. Unfortunately, today most women with PKU have discontinued dietary treatment by adulthood and find the restricted phe diet onerous and difficult. Fortunately, this is changing. "Diet for life" is the usual, although not yet universally, adopted practice today, but even so, there are women who conceive "off diet." This inhibits intellectual development of the fetus. If intensive services are provided for such women, fetal outcome can be improved by good blood phe control between 120 and 360 uM/L. Although prenatal diagnosis is available by fetal mutation studies, many women today resist the benefits of genetic counseling. Unfortunately, insurance companies often are unwilling to pay for such procedures as mutation analysis or the provision of low phe diets. Overall, public policy for the care of women with genetic disorders is in a state of flux and strong leadership is required to improve services. PMID- 11499063 TI - Postgenomic medicine. Presymptomatic testing for prediction and prevention. AB - Significant changes are occurring in genetic screening paradigms. Genetic screening is moving from traditional analytes, such as small molecules and proteins, to molecular genetic testing involving DNA and RNA. There are significant consequences to these changes, involving issues for the family unit, such as misattribution of parentage, and concerns regarding discrimination, confidentiality, and privacy. Although these latter issues have broader concerns for medicine and medical information, in the context of genetic testing, information derived from one individual can have a significant impact on others within their family. Screening is also changing from mendelian disease ascertainment to predictive testing. Issues that arise involve appropriate age at testing for adult-onset disorders, the clinical validity and clinical use of genetic testing for complex diseases, and the efficacy of interventions following genetic testing. We are also learning that the phenotypes of even simple mendelian disorders are influenced by complex genetic and environmental factors. The observations that genotypes rarely predict phenotypes absolutely have significant ramifications for counseling based on mutation analysis, for example in neonates who have not yet manifested symptoms and in older children and in adults undergoing predictive testing. Molecular genetic testing often proceeds rapidly from the research laboratory to the clinical setting. We must recognize that for single-gene disorders with high penetrance, the information derived from such testing may be relatively easy to interpret and apply. For complex diseases, however, the populations studied and their demographic characteristics are extremely important for extrapolation to counseling of individual patients. The value of population-based predictive testing is exemplified by newborn screening. It is clear that the Human Genome Project, and the information and technologies from it, will have a much broader impact on public health by presymptomatic prediction and prevention of disease. PMID- 11499064 TI - Sickle cell. AB - The initiation of newborn screening and its virtually universal implementation will eventually yield a population in which sickle cell disease has been identified and comprehensive care is provided for children. The situation with SCT is different; there will continue to be the identification of parents who have the potential for having a child with a sickle cell disease but because they will not be tested or counseled, there will continue to be a population of children with a sickle cell disease whose parents have not been enabled to make informed decisions that they believe are in their best interest relative to family planning. Also, we will continue to have a population of pregnant women with a fetus with sickle cell disease who will not be given an opportunity to decide whether they wish to continue or terminate the pregnancy. They all will give birth to a child with a lifetime of chronic illness with its associated psychological, social, and financial burdens for the individual and his or her parents. The failure to implement prenatal diagnosis is an abridgment of two fundamental rights: the right to know and the right to decide. In this case it is the right to know about the potential health status of their children if that is possible, and the right to decide about the actual health care status of their children if options are available. PMID- 11499065 TI - Neonatal screening for metabolic, endocrine, infectious, and genetic disorders. Current and future directions. AB - There are good reasons to expect that future neonatal screening will expand both to include more disorders and to cover more of the global newborn population. Disorders for which neonatal screening will be given high priority in the health care field in the future are CH and PKU. Screening for CH is likely to expand faster than screening for PKU, especially in the developing world. In the future, screening for CAH will be practiced much more widely than today. Screening for CF is likely to qualify for routine neonatal screening in the future, especially if gene therapy becomes successful. Screening for infectious diseases is an area that is also developing rapidly. Which disorders to screen for neonatally will depend on a number of factors that are unique to each society, such as the prevalence, economy, and ethics. This must be realized when international guidelines are drafted. Technical development, which is of major importance for neonatal screening, includes MS-MS, different DNA techniques, and automation. The expansion of biomedical knowledge in a wide variety of fields will establish new grounds for neonatal screening. PMID- 11499066 TI - Medicolegal genetic issues. AB - As genetic advances are made at an increasingly rapid rate, the physician practicing in this specialty must be aware of the most current data, risks, and options for treatment to comply with the standard of care. Only by arming himself or herself with this information, imparting it to the patient, and appropriately documenting that the patient has been informed, can today's practitioner expect to stay abreast of legal liability and the potentially enormous damages accompanying genetic lawsuits. PMID- 11499067 TI - Future directions. AB - With the continued explosion of genetic technology, the number of disorders amenable to screening is expanding geometrically. Historically, most genetic screening has been in the newborn period. Much more can be done for the fetus if genetic screening and diagnosis are accomplished early in the pregnancy rather than after birth. The principal requirements to make neonatal screening disorders possible in the first trimester center around those tests that can be one on a molecular basis, and the development of fetal cell isolation from the maternal circulation. Over the course of the next decade, it is likely that many of the tests currently performed in the newborn period will be accomplished in the early or mid-gestational period. PMID- 11499068 TI - The full cost of dental insurance: who's really minding the store? AB - A private practitioner describes the evolution of control of practice by insurance companies. The effects of assignment of benefits have been to limit patients' demand for care to what will be covered by insurance, postponement of needed care, damaging the relationship between patients and dentists, and erosion of patients' sense of responsibility for their own oral health. Current trends toward electronic filing of insurance claims are seen as an acceleration of these practices. Educating patients regarding their role in their own care is proposed as a needed response. PMID- 11499069 TI - Dental benefits--expanding access to dental care. AB - The Executive Director of the National Association of Dental Plans presents statistics describing recent trends in product mix, growth in voluntary benefits and referral plans, geographic concentration, and industry consolidation in the dental benefits industry. Current issues include dental workforce, the economy, human resources policies, and regulation. The issue of quality oral health care is identified as needing consensus definition by the entire dental industry. PMID- 11499070 TI - The case for dental preferred provider organizations. AB - The dental PPO market is the fastest growing segment of the dental benefits business. As with traditional indemnity plans, dentists are reimbursed on a fee for-service basis. Members can refer themselves to dentists of choice, including specialists, inside and outside the network. Employers' interest in DPPOs is expected to continue as they seek to control or reduce expenses associated with dental benefits plans. Dentist participation in managed care programs significantly lags medical, and fewer than half of practicing dentists participate in DPPOs. Negotiated discounts vary across carriers and geographic regions, generally ranging 15% to 30% off average fees. The American Dental Association predicts that dentists' participation in DPPOs will continue to increase, indicating further growth for these dental plans. There are financial incentives for members to stay in network. PMID- 11499071 TI - A position paper: six lessons about managed care in dentistry. AB - The limitation of resources available for oral health care makes it inevitable that groups (such as third parties) would arise with a view toward conservation of these resources. Although the concept of adequate care has been generally operationally defined in dentistry, a confusion remains between adequate and best. Dentists may be under an ethical obligation to inform patients of all available treatment options (ranging from the adequate to the best) and to perform the selected treatment to the best of their ability, but they are not under an obligation to offer only the best alternative or to expect that society will make the resources available for that level of care. PMID- 11499072 TI - Benchmarking. PMID- 11499073 TI - Dental prepayment: the past, present, and the future. AB - Insurance now pays for slightly less than 50% of America's dental bill. The explosive growth in dental insurance in the 1970s and 1980s is traced to the tax effect (coverage paid for with pre-tax dollars offsets fees up to a point) and the insurance effect (costs for care can be projected, although improvements in oral health status are leading to lower projected costs). An equilibrium point appears to have been reached. Dentists must weigh the trade-offs between discounted fees and increased number of patients, and carriers must weight the trade-offs between smaller discounts and wider participation by dentists. There are no market forces forecast that will substantially change this equilibrium in the near future. PMID- 11499074 TI - Dental insurance: design, need, and public policy. AB - The demand for dental insurance is likely to increase as our nation ages. However, future dental benefit plans may need to differ considerably from their present day counterparts to be cost-effective. These plans will be designed to minimize adverse selection, limiting or excluding some components found in today's dental insurance plans. Interest in improving access to care for the undeserved has gained significant support as of late. Dentistry should be prepared to provide the leadership necessary to help shape the design of future dental plans and to help improve the effectiveness of public coverage programs. PMID- 11499075 TI - PACS--just another casino game? PMID- 11499076 TI - Radiology managers benefit when GPOs and vendors collaborate. PMID- 11499077 TI - Lies, damn lies, and statistics. PMID- 11499078 TI - Delegation: developing the habit. AB - Often, individuals take personal delegation skills for granted and assume the presence of expertise with the practice of delegation, which may not be the case. Those assumptions can be found at both ends of the process, with the manager and the employee. Every time a manager places an employee in a job and gives him or her a job description or a set of instructions, the manager has delegated. The manager has placed someone in a position to perform operations for which ultimately the manager is responsible. Delegation is both a process and a condition. The process is the act of assigning work to an employee; the condition of delegating a job is a thorough and mutual understanding between the supervisor and the employee of specific results and methods by which these results can be achieved. The condition goes far beyond the simple process of assigning a job. The point at which many managers fail in delegating is in neglecting to move past the process and take the required steps to establish a true condition of delegation. Failure to delegate is the leading cause of managers retarding their professional growth. In the case of a workaholic--someone who fails to learn the value of delegation--the job soon becomes too much, and the effectiveness of the department may suffer. By reducing the burden of technical duties and busy work, managers will find that it is possible to be more effective and actually spend more time managing. A number of the reasons why managers fail to delegate are complex and subconscious, such as insecurity, fear of competition and even fear of not being recognized for accomplishments achieved. Other reasons for failing to delegate are habit and shortages of staff members or time. Delegation is an investment in time. The eventual gain from such an investment, which may temporarily cause the department to fall further behind during a training period, outweighs the costs. The manager is the final authority in such duties as approval, recommendations and implementations. Remember that to delegate authority does not mean to delegate ultimate responsibility. Only the manager should bear the burden of responsibilities that directly affect his or her career. One of the most common mistakes in the process of delegating is to turn an employee loose on a job with inadequate instruction. Too often this is caused by lack of time. Strictly speaking, when managers assign tasks to employees, they are delegating the authority to perform the task. However, all responsibility for the completion of the task remains with the manager. In most instances, responsibility for the failure of a delegated project lies with the manager, not the employee. While some failure is to be expected, it can be minimized by proper evaluation of the situation and proper communication. Delegation is a calculated risk. The manager should be willing to take a risk to see if an employee does have the skills and ability to perform the specific task. How else will the manager ever know? Developing a strong habit of delegation will lead to development of an effective and efficient staff and department. Delegation will serve not only the organization well, but also managers throughout their careers. PMID- 11499079 TI - Addressing the shortage of radiologists. AB - In a survey of 254 hospital radiology departments conducted last year, Dallas based U.S. Radiology Partners (USRP), a radiology management firm, found that 45 percent of hospitals are understaffed in radiology. Fifty-six percent of hospital radiology department heads surveyed by USRP indicated that staffing shortages were diminishing the quality of care their departments are able to provide. Moreover, staffing shortages are occurring at a time when radiology volume generally is increasing. There are various underlying reasons why the supply of radiologists and other physicians is insufficient to meet demand in many areas, including institutional misdiagnosis, graduate medical education, patient preferences, the economy, restrictions on international medical graduates, practice patterns, patient and physician demographics and new technology. Given current physician supply and demand trends, a strategic radiology staffing plan is needed. Such a plan would include the following elements: retention, candidate parameters, contracts and incentives, sourcing, screening, interviewing, responsiveness and decisiveness. PMID- 11499080 TI - Radiology system evolution in the new millennium. AB - For many decades the practice of radiology grew slowly in America and was largely a secondary function under the control of hospitals. In more recent times it has vastly expanded its array of diagnostic, interventional, and therapeutic abilities. There is increasing consumer logic for direct access. Motivations have grown to create large independent entities with broadly diverse capabilities in order to succeed in the new millennium. Most regional markets are evolving rapidly in terms of managed care penetration, health system formation, physician practice consolidation and aggressive purchaser behavior by employers and consumers. To understand the enormity of healthcare evolution, it is useful to look at the industry's paradigm shifts in recent decades. Virtually every aspect of organizational infrastructure, delivery approaches, and the business environment has evolved markedly during the past fifty years. These changes will accelerate. To succeed financially, radiology groups must strengthen their market positions, technical capabilities, continuums of care and geographic dominance. Equally important is the wisdom of diversifying incomes into related services and businesses that provide additional related revenues. Key factors for successful development include facility market growth, full coverage of managed care contracts, high efficiency and aggressive diversification. A fully evolved system generates significant revenues and profitability by protecting and strengthening its financial position in this environment. That is accomplished through the development of strategically located radiology groups, aggressive alliances with medical practices in allied disciplines, and managed radiology departments and facilities for partner health systems. Organizational success ultimately depends on the ability to accept capitated payments under risk-bearing arrangements. The strategic business plan should be organized with the appropriate levels of detail needed to establish executive focus and priorities. These should be woven into operational and capital budgets to reflect expectations of the revenues, expenses and investments tied to the plan. While formidable, all of these objectives are realistic and can be accomplished if the right decisions are made. Initially, the entity's principle business objective is to formulate and begin implementation of methodical yet aggressive strategies for growth that are sensitive to sustaining high levels of quality patient care. The next phase features mergers with large, independent radiology practices in key geographic areas and successful acquisition of smaller practices. The objective of the final phase is to aggressively expand into select metropolitan areas with regional coverage and full teleradiology capabilities. High levels of market strength and financial performance are necessary to succeed. Passive limitations to small geographic areas and narrow practices will undermine their market position and dissolve financial strength with no hope of recovery. Only the dominant systems will survive and prosper. PMID- 11499081 TI - Practical solutions for staff recruitment & retention. AB - There are three essential topics for radiology managers to consider in light of persistent staffing shortages: support of the profession and educational programs, perks as recruitment tools and incentives as retention tools. Some activities that can help support departments and educational programs for radiologic technologists are job shadowing, training for volunteer services, advanced placement for school applicants, sponsoring an educational program or clinical training site, creating a positive work environment and supporting outreach projects geared to local high schools. Traditional perks used in recruitment efforts have included relocation assistance, travel and lodging expenses during the interview process, loan repayment, scholarships and sign-on bonuses. Some common incentives for retaining employees are tuition reimbursement, cross training, availability of educational resources, continuing education opportunities, professional development and incremental increases in salary. There are many other tools that can be used, such as career ladders, creating an environment conducive to teamwork or a more personal atmosphere and showcasing talents of various staff members. There is much overlap among these suggestions in support of the profession and educational programs, recruitment and retention of qualified staff radiologic technologists. Radiology managers can and should be creative in developing different programs to build loyalty and commitment to a radiology department. PMID- 11499082 TI - MRI technology overview. PMID- 11499083 TI - Change? What change? PMID- 11499084 TI - Following the force. PMID- 11499085 TI - Going public on physician liability: what difference does it make? AB - States are spending time and money to collect and post on the Internet information about a physicians medical malpractice history. Is anyone listening? PMID- 11499086 TI - California hospital stakes a claim in growing use of artificial intelligence. AB - John Muir-Mt. Diablo Health Network in northern California has found the perfect employee--RoboCOPS, an automated program from IntelliClaim, the Norwalk, CT-based claims performance company. PMID- 11499087 TI - Hospital looks for data security, also gets improved productivity. AB - What started as a way to protect computerized patient information from unauthorized users turned into a major cost-cutting initiative for Houston's Memorial Hermann Hospital. Administrators describe why they're expanding the security initiative systemwide. PMID- 11499088 TI - New software tool allows physician groups to benchmark key clinical measures. AB - An innovative software tool launched by a partnership between MGMA a and South Carolina software vendor is promising to give physician groups unparalleled benchmarking data on practice patterns and productivity. PMID- 11499089 TI - Risk adjustment drives statewide benchmarking effort. PMID- 11499090 TI - Post-rape care in hospital emergency rooms. PMID- 11499091 TI - Readers respond to "Cholera in Paris". PMID- 11499092 TI - Readers respond to "Cholera in Paris". PMID- 11499093 TI - Global environmental change as "risk factor": can epidemiology cope? PMID- 11499094 TI - Cautiously adjusting to the new millennium: changing to the 2000 population standard. PMID- 11499095 TI - Can unsafe sex behind bars be barred? PMID- 11499096 TI - Using technology to advance the public's health. PMID- 11499097 TI - Health information systems and health communications: narrowband and broadband technologies as core public health competencies. PMID- 11499098 TI - Promoting public health through electronic media: a challenge for schools of public health. PMID- 11499099 TI - Bringing the mountain to Mohammed: a mobile dental team serves a community-based program for people with HIV/AIDS. AB - In spite of the direct referral system and family-centered model of primary oral health care linking medical and dental care providers, most HIV-positive patients at the Columbia Presbyterian Medical Center received only emergency and episodic dental care between 1993 and 1998. To improve access to dental care for HIV/AIDS patients, a mobile program, called WE CARE, was developed and colocated in community-based organizations serving HIV-infected people. WE CARE provided preventive, early intervention, and comprehensive oral health services to minorities, low-income women and children, homeless youths, gays and lesbians, transgender individuals, and victims of past abuse. More efforts to colocate dental services with HIV/AIDS care at community-based organizations are urgently needed. PMID- 11499100 TI - "...so that others may walk": the March of Dimes. PMID- 11499101 TI - Is child health at risk while families wait for housing vouchers? PMID- 11499102 TI - Encouraging stair use: stair-riser banners are better than posters. PMID- 11499103 TI - The association between extreme precipitation and waterborne disease outbreaks in the United States, 1948-1994. AB - OBJECTIVES: Rainfall and runoff have been implicated in site-specific waterborne disease outbreaks. Because upward trends in heavy precipitation in the United States are projected to increase with climate change, this study sought to quantify the relationship between precipitation and disease outbreaks. METHODS: The US Environmental Protection Agency waterborne disease database, totaling 548 reported outbreaks from 1948 through 1994, and precipitation data of the National Climatic Data Center were used to analyze the relationship between precipitation and waterborne diseases. Analyses were at the watershed level, stratified by groundwater and surface water contamination and controlled for effects due to season and hydrologic region. A Monte Carlo version of the Fisher exact test was used to test for statistical significance. RESULTS: Fifty-one percent of waterborne disease outbreaks were preceded by precipitation events above the 90th percentile (P = .002), and 68% by events above the 80th percentile (P = .001). Outbreaks due to surface water contamination showed the strongest association with extreme precipitation during the month of the outbreak; a 2-month lag applied to groundwater contamination events. CONCLUSIONS: The statistically significant association found between rainfall and disease in the United States is important for water managers, public health officials, and risk assessors of future climate change. PMID- 11499104 TI - Association of normal weather periods and El Nino events with hospitalization for viral pneumonia in females: California, 1983-1998. AB - OBJECTIVES: This study examined associations between weather and hospitalizations of females for viral pneumonia during normal weather periods and El Nino events in the California counties of Sacramento and Yolo, San Francisco and San Mateo, and Los Angeles and Orange. METHODS: Associations between weather and hospitalizations (lagged 7 days) for January 1983 through June 1998 were evaluated with Poisson regression models. Generalized estimating equations were used to adjust for autocorrelation and overdispersion. Data were summed over 4 days. RESULTS: Associations varied by region. Hospitalizations in San Francisco and Los Angeles increased significantly (30%-50%) with a 5 degrees F decrease in minimum temperature. Hospitalizations in Sacramento increased significantly (25% 40%) with a 5 degrees F decrease in maximum temperature difference. The associations were independent of season. El Nino events were associated with hospitalizations only in Sacramento, with significant decreases for girls and increases for women. CONCLUSIONS: The results suggest that viral pneumonia could continue to be a major public health issue, with a significant association between weather and hospitalizations, even as the global mean temperature continues to rise. An understanding of population sensitivity under different weather conditions could lead to an improved understanding of virus transmission. PMID- 11499105 TI - Changing to the 2000 standard million: are declining racial/ethnic and socioeconomic inequalities in health real progress or statistical illusion? AB - OBJECTIVES: This study determined the effects of changing from the 1940 to the 2000 standard million on monitoring socioeconomic and racial/ethnic inequalities in health. METHODS: Using the 1940, 1970, and 2000 standard million, we calculated and compared age-adjusted rates for selected health outcomes stratified by socioeconomic level. RESULTS: Changing from the 1940 to the 2000 standard million markedly reduced the age-adjusted relative risks for self reported fair or poor health status of poor Americans compared with high-income Americans. CONCLUSIONS: Public health researchers and practitioners should give serious consideration to the implications of the change to the 2000 standard million for monitoring social inequalities in health. PMID- 11499106 TI - Retrospective validation of a surveillance system for unexplained illness and death: New Haven County, Connecticut. AB - OBJECTIVES: This study investigated retrospective validation of a prospective surveillance system for unexplained illness and death due to possibly infectious causes. METHODS: A computerized search of hospital discharge data identified patients with potential unexplained illness and death due to possibly infectious causes. Medical records for such patients were reviewed for satisfaction of study criteria. Cases identified retrospectively were combined with prospectively identified cases to form a reference population against which sensitivity could be measured. RESULTS: Retrospective validation was 41% sensitive, whereas prospective surveillance was 73% sensitive. The annual incidence of unexplained illness and death due to possibly infectious causes during 1995 and 1996 in the study county was conservatively estimated to range from 2.7 to 6.2 per 100,000 residents aged 1 to 49 years. CONCLUSIONS: Active prospective surveillance for unexplained illness and death due to possibly infectious causes is more sensitive than retrospective surveillance conducted through a published list of indicator codes. However, retrospective surveillance can be a feasible and much less labor intensive alternative to active prospective surveillance when the latter is not possible or desired. PMID- 11499107 TI - An outbreak of syphilis in Alabama prisons: correctional health policy and communicable disease control. AB - OBJECTIVES: After syphilis outbreaks were reported at 3 Alabama State men's prisons in early 1999, we conducted an investigation to evaluate risk factors for syphilis infection and describe patterns of syphilis transmission. METHODS: We reviewed medical, patient interview, and prison transfer records and documented sexual networks. Presumptive source cases were identified. Odds of exposure to unscreened jail populations and transfer from other prisons were calculated for case patients at 1 prison. RESULTS: Thirty-nine case patients with early syphilis were identified from 3 prisons. Recent jail exposure (odds ratio [OR] = 8.0, 95% confidence interval [CI] = 0.3, 158.7, P = .14) and prison transfer (OR = 32.0, 95% CI = 1.6, 1668.1, P < .01) were associated with being a source case patient. CONCLUSIONS: Probable sources of syphilis introduction into and transmission within prisons included mixing of prisoners with unscreened jail populations, transfer of infected inmates between prisons, and multiple concurrent sexual partnerships. Reducing sexual transmission of disease in correctional settings is a public health priority and will require innovative prevention strategies. PMID- 11499108 TI - Increasing voluntary HIV testing by offering oral tests in incarcerated populations. AB - OBJECTIVES: This study reports responses of incarcerated persons to voluntary blood and oral HIV testing. METHODS: Males and females in local detention and juvenile justice facilities in Maryland (n = 1314) chose oral or blood testing and reported reactions to the oral HIV test. The relationship of demographics and HIV risk factors to test choice was examined. RESULTS: Reactions to oral testing were very favorable; some participants reported that they would not otherwise have been tested. Participants who chose oral testing were more likely to be male and African American, but they did not differ from those who chose blood testing in most risk factors or in seroprevalence. CONCLUSIONS: Oral HIV testing in correctional settings may promote voluntary testing among persons who otherwise would refuse or avoid testing, especially among groups (males and African Americans) disproportionately affected by HIV. PMID- 11499109 TI - Continuing-education needs of the currently employed public health education workforce. AB - OBJECTIVES: This study examined the continuing-education needs of the currently employed public health education workforce. METHODS: A national consensus panel of leading health educators from public health agencies, academic institutions, and professional organizations was convened to examine the forces creating the context for the work of public health educators and the competencies they need to practice effectively. RESULTS: Advocacy; business management and finance; communication; community health planning and development, coalition building, and leadership; computing and technology; cultural competency; evaluation; and strategic planning were identified as areas of critical competence. CONCLUSIONS: Continuing education must strengthen a broad range of critical competencies and skills if we are to ensure the further development and effectiveness of the public health education workforce. PMID- 11499110 TI - A conceptual framework to measure performance of the public health system. AB - OBJECTIVES: This article describes a unifying conceptual framework for the public health system as a way to facilitate the measurement of public health system performance. METHODS: A conceptual framework for the public health system was developed on the basis of the work of Donabedian and a conceptual model previously developed by Bernard Turnock and Arden Handler. RESULTS: The conceptual framework consists of 5 components that can be considered in relationship to each other: macro context, mission, structural capacity, processes, and outcomes. Although the availability of measures for each of these components varies, the framework can be used to examine the performance of public health systems as well as that of agencies and programs. CONCLUSIONS: A conceptual framework that explicates the relationships among the various components of the public health system is an essential step toward providing a science base for the study of public health system performance. PMID- 11499111 TI - Accessibility of primary care services in safety net clinics in New York City. AB - OBJECTIVES: This study analyzed data from a survey of New York City ambulatory care facilities to determine primary care accessibility for low-income patients, as evidenced by the availability of enabling services, after-hours coverage, and policies for serving the uninsured. METHODS: Ambulatory care facilities were surveyed in 1997, and analysis was performed on a set of measures related to access to care. Only sites that provided comprehensive primary care services were included in the analysis. For comparison, site were classified by sponsorship (public, nonprofit voluntary hospital, federally qualified health center, non hospital-sponsored community health center). RESULTS: Publicly sponsored sites and federally qualified health center sites showed the strongest performance across nearly all the measures of accessibility that were examined. CONCLUSIONS: As safety net clinics confront the financial strain of implementing mandatory Medicaid managed care while also dealing with declining Medicaid caseloads and increasing numbers of uninsured, their ability to sustain the policies and services that support primary care accessibility may be threatened. PMID- 11499112 TI - The effect of primary care physician supply and income inequality on mortality among blacks and whites in US metropolitan areas. AB - OBJECTIVES: This study assessed whether income inequality and primary care physician supply have a different effect on mortality among Blacks compared with Whites. METHODS: We conducted a multivariate ecologic analysis of 1990 data from 273 US metropolitan areas. RESULTS: Both income inequality and primary care physician supply were significantly associated with White mortality (P < .01). After the inclusion of the socioeconomic status covariates, the effect of income inequality on Black mortality remained significant (P < .01), but the effect of primary care physician supply was no longer significant (P > .10), particularly in areas with high income inequality. CONCLUSIONS: Improvement in population health requires addressing socioeconomic determinants of health, including income inequality and primary care availability and access. PMID- 11499113 TI - Black-white differences in infectious disease mortality in the United States. AB - OBJECTIVES: This study determined the degree to which Black-White differences in infectious disease mortality are explained by income and education and the extent to which infectious diseases contribute to Black-White differences in all-cause mortality. METHODS: A sample population of the National Longitudinal Mortality Study from 1979 through 1981 was analyzed and followed up through 1989. RESULTS: Infectious disease mortality among Blacks was higher than among Whites, with a relative risk of 1.53 after adjustment for age and sex and 1.34 after further adjustment for income and education. Death from infectious diseases contributed to 9.3% of the difference in all-cause mortality. CONCLUSIONS: In the United States, infectious diseases account for nearly 10% of the excess all-cause mortality rates in Blacks compared with Whites. PMID- 11499114 TI - Acculturation and leisure-time physical inactivity in Mexican American adults: results from NHANES III, 1988-1994. AB - OBJECTIVES: This study examined the relationship between acculturation and leisure-time physical inactivity among Mexican American adults. METHODS: Using data from the Third National Health and Nutrition Examination Survey, we estimated the prevalence of physical inactivity according to place of birth and language used at home. RESULTS: Spanish-speaking Mexican Americans had a higher prevalence of physical inactivity during leisure time than those who spoke mostly English, independent of place of birth. CONCLUSIONS: Acculturation seems to be positively associated with participation in leisure-time physical activity. PMID- 11499115 TI - Heart rate as a predictor of mortality: the MATISS project. AB - OBJECTIVES: This study sought to verify the independent role of heart rate in the prediction of all-cause, cardiovascular, and noncardiovascular mortality in a low risk male population. METHODS: In an Italian population-based observational study, heart rate was measured in 2533 men, aged 40 to 69 years, between 1984 and 1993. Data on cardiovascular risk factors were collected according to standardized procedures. Vital status was updated to December 1997. RESULTS: Of 2533 men followed up (representing 24,457 person-years), 393 men died. Age adjusted death rates for 5 heart rate levels showed increasing trends. The adjusted hazard rate ratios for each heart rate increment were 1.52 (95% confidence interval [CI] = 1.29, 1.78) for all-cause mortality, 1.63 (95% CI = 1.26, 2.10) for cardiovascular mortality, and 1.47 (95% CI = 1.19, 1.80) for noncardiovascular mortality. Relative risks between extreme levels were more than 2-fold for all endpoints considered. CONCLUSIONS: Heart rate is an independent predictor of cardiovascular, noncardiovascular, and total mortality in this Italian middle-aged male population. PMID- 11499116 TI - Colorectal cancer screening participation: comparisons with mammography and prostate-specific antigen screening. AB - OBJECTIVES: The relation of personal characteristics, health and lifestyle behaviors, and cancer screening practices to current colorectal cancer (CRC) screening was assessed and compared with those factors' relation to current mammography screening in women and prostate-specific antigen (PSA) screening in men. METHODS: A cross-sectional random-digit-dialed telephone survey of 954 Massachusetts residents aged 50 and older was conducted. RESULTS: The overall prevalence of current CRC screening was 55.3%. Logistic regression results indicated that family history of CRC (odds ratio [OR] = 1.98; 95% confidence interval [CI] = 1.02, 3.86), receiving a regular medical checkup (OR = 3.07; 95% CI = 2.00, 4.71), current screening by mammography in women and PSA in men (OR = 4.40; 95% CI = 2.94, 6.58), and vitamin supplement use (OR = 1.87; 95% CI = 1.27, 2.77) were significant predictors of CRC screening. CONCLUSIONS: Health and lifestyle behaviors were related to increased current CRC, mammography, and PSA screening. Personal factors independently related to CRC screening were not consistent with those related to mammography and PSA screening. This lack of consistency may reflect different stages of adoption of each type of screening by clinicians and the public. PMID- 11499117 TI - The effect of different definitions of a patient on immunization assessment. AB - OBJECTIVES: In this report, the authors compare immunization assessment using 2 definitions of a patient. METHODS: Two Clinical Assessment Software Application (CASA) assessments were performed. The first sampled 200 two-year-olds seen at least once since birth. The second sampled 200 two-year-olds seen in the previous year. Children with incomplete immunizations in the first sample were contacted. RESULTS: In the second sample, 72% of children had complete immunizations, compared with 46% in the first sample. In the first sample, 78% of children with incomplete immunizations had not been seen during the past year. Of 134 children in the first sample seen in the past year or successfully contacted, 75% had complete immunizations. CONCLUSIONS: The CASA assessment's definition of a patient underestimates immunization rates. PMID- 11499118 TI - Adolescent sexual orientation and suicide risk: evidence from a national study. AB - OBJECTIVES: Sexual orientation has been a debated risk factor for adolescent suicidality over the past 20 years. This study examined the link between sexual orientation and suicidality, using data that are nationally representative and that include other critical youth suicide risk factors. METHODS: Data from the National Longitudinal Study of Adolescent Health were examined. Survey logistic regression was used to control for sample design effects. RESULTS: There is a strong link between adolescent sexual orientation and suicidal thoughts and behaviors. The strong effect of sexual orientation on suicidal thoughts is mediated by critical youth suicide risk factors, including depression, hopelessness, alcohol abuse, recent suicide attempts by a peer or a family member, and experiences of victimization. CONCLUSIONS: The findings provide strong evidence that sexual minority youths are more likely than their peers to think about and attempt suicide. PMID- 11499119 TI - Beyond assumptions of negligible risk: sexually transmitted diseases and women who have sex with women. AB - OBJECTIVES: This study evaluated the association of female-female sexual behavior with sexually transmitted diseases (STDs). METHODS: Female participants (n = 286) were recruited from the Twin Cities Gay/Lesbian/Bisexual/Transgender Pride Festival. Logistic regression was used to examine the association between female female sexual behavior and STDs. RESULTS: Women in all partner history groups, including 13% of women with only female partners, reported a history of STD. Increased sexual exposures with women predicted an increase in the likelihood of STDs after known risk factors had been controlled. Neither number of female partners nor number of exposures was associated with obtaining regular STD testing. CONCLUSIONS: The risk of STDs through female-female sexual exposure is not negligible. Nevertheless, patterns of STD testing do not reflect this risk. PMID- 11499120 TI - Prevalence of genital chlamydial infection in young women entering a national job training program, 1990-1997. AB - OBJECTIVES: This analysis describes trends in the prevalence of genital chlamydial infection in economically disadvantaged young women entering a national job training program. METHODS: We examined chlamydia test data for May 1990 through June 1997 for women aged 16 to 24 years who enrolled in the program. The significance of trends was evaluated with the chi 2 test for trend. RESULTS: Prevalence of chlamydial infection declined 32.9%, from 14.9% in 1990 to 10.0% in 1997 (P < .001). Prevalence decreased significantly in all age groups, racial/ethnic groups, and geographic regions. CONCLUSIONS: The decrease in prevalence of chlamydial infection suggests that prevention activities have reached disadvantaged women across the United States; however, prevalence of chlamydial infection remains high, and enhanced prevention efforts in disadvantaged communities are urgently needed. PMID- 11499121 TI - Trends in HIV testing among pregnant women: United States, 1994-1999. AB - OBJECTIVES: This study evaluated 1995 guidelines for HIV testing of pregnant women. METHODS: Analysis focused on Behavioral Risk Factor Surveillance System data for the years 1994 through 1999. Data were aggregated across states. RESULTS: Percentages of pregnant women tested for HIV increased from 1995 to 1996 (from 41% to 53%) and again from 1997 (52%) to 1998 (60%). CONCLUSIONS: After implementation of the guidelines, the percentage of pregnant women tested for HIV increased, although nearly half had not been tested. More efforts are needed to encourage women to undergo testing for HIV during pregnancy, thus maximizing opportunities for offering antiretroviral therapy. PMID- 11499122 TI - An intervention for parents with AIDS and their adolescent children. AB - OBJECTIVES: This study evaluated an intervention designed to improve behavioral and mental health outcomes among adolescents and their parents with AIDS. METHODS: Parents with AIDS (n = 307) and their adolescent children (n = 412) were randomly assigned to an intensive intervention or a standard care control condition. Ninety-five percent of subjects were reassessed at least once annually over 2 years. RESULTS: Adolescents in the intensive intervention condition reported significantly lower levels of emotional distress, of multiple problem behaviors, of conduct problems, and of family-related stressors and higher levels of self-esteem than adolescents in the standard care condition. Parents with AIDS in the intervention condition also reported significantly lower levels of emotional distress and multiple problem behaviors. Coping style, levels of disclosure regarding serostatus, and formation of legal custody plans were similar across intervention conditions. CONCLUSIONS: Interventions can reduce the long-term impact of parents' HIV status on themselves and their children. PMID- 11499123 TI - Challenges associated with increased survival among parents living with HIV. AB - OBJECTIVES: This study examined sociodemographic and psychosocial factors that predict survival among parents living with HIV. METHODS: Parents with HIV (n = 307) were recruited from 1993 to 1995 in New York City and repeatedly assessed. Survival was monitored among the sample (81% mothers; 45% Latino, 34% African American). RESULTS: Over a median period of 28 months (range = 0-53 months), 44% (n = 135) of the parents died. Having an AIDS diagnosis and being African American were associated with earlier death. Sex, age, and financial status were not related to survival. Parents who survived had initially higher levels of anxiety that decreased over time; in contrast, parents who died reported initially lower, but constant, levels of anxiety over time. After HIV diagnostic status was controlled for, it was found that parents who reported having more children, using a coping style of seeking social support, and being sexually active at baseline survived longer. CONCLUSIONS: The counterintuitive findings raise hypotheses regarding the role of change and responsibilities in the survival of parents with HIV. PMID- 11499125 TI - Physicians, vendors and HIPAA compliance. PMID- 11499124 TI - Brief intervention for heavy-drinking college students: 4-year follow-up and natural history. AB - OBJECTIVES: This study examined long-term response to an individual preventive intervention for high-risk college drinkers relative to the natural history of college drinking. METHODS: A single-session, individualized preventive intervention was evaluated within a randomized controlled trial with college freshmen who reported drinking heavily while in high school. An additional group randomly selected from the entire screening pool provided a normative comparison. Participant self-report was assessed annually for 4 years. RESULTS: High-risk controls showed secular trends for reduced drinking quantity and negative consequences without changes in drinking frequency. Those receiving the brief preventive intervention reported significant additional reductions, particularly with respect to negative consequences. Categorical individual change analyses show that remission is normative, and they suggest that participants receiving the brief intervention are more likely to improve and less likely to worsen regarding negative drinking consequences. CONCLUSIONS: Brief individual preventive interventions for high-risk college drinkers can achieve long-term benefits even in the context of maturational trends. PMID- 11499126 TI - Deploying wireless LANs. Today's smart planning helps wireless users to add and expand well beyond tomorrow. PMID- 11499127 TI - Winning with wireless. Avoiding the shock of the unplugged world. PMID- 11499128 TI - Walk a wireless mile. Research reveals big divide in expectations versus experiences among healthcare providers. PMID- 11499129 TI - Nursing system makes a difference. Northwest IDS reaps the benefits of using automated tools to support nursing. PMID- 11499130 TI - Connection tops collection. Peer-to-peer technology lets caregivers access necessary data, upon request, without using a repository. PMID- 11499131 TI - Truth or consequences. Credentialing is vital for the health of a hospital's patients, and ultimately, the hospital itself. PMID- 11499132 TI - Getting it right for providers. Louisiana-based health plan successfully puts managed care providers online and efficiency at the top of everyone's list. PMID- 11499133 TI - Automated billing systems and compliance. Nurses must become knowledgeable about software systems that can and cannot help eliminate errors. PMID- 11499134 TI - Where medicine and technology meet. Four strategies for gaining physician support and acceptance of evidence-based medicine. PMID- 11499136 TI - Wireless hotlist. PMID- 11499135 TI - What works: scheduling. Picture perfect solution. The right technology and an ASP solution bring scheduling efficiency and added revenue to a community hospital's radiology department. PMID- 11499137 TI - One physician's journey into e-success. PMID- 11499138 TI - [Therapy of hypoparathyroidism with parathyroid hormone injections? When, then only for problem cases]. PMID- 11499139 TI - [US study for prevention of Lyme borreliosis. An antibiotic after every tick bite? (interview by Dr. Beate Schumacher)]. PMID- 11499140 TI - [Risk of a diabetes epidemic. This is how you can stop it!]. PMID- 11499141 TI - [Pneumococcal infections. New vaccine protects even young children]. PMID- 11499142 TI - [What is effective in traveler's diarrhea? Current recommendations on prevention, therapy and after care]. AB - Traveller's diarrhea is almost always due to an infection with intestinal bacteria, viruses or parasites. The leading agents are the ubiquitous enterotoxic E. coli bacteria. Prophylaxis is achieved primarily by the strict avoidance of fecally contaminated food and drink--advice which, however, is often ignored. Chemoprophylactic measures include the use of probiotics, in particular Saccharomyces boulardii, with antibiotics being given only in special cases. Treatment consists mainly in the replacement of lost fluids and electrolytes in the form of oral rehydration solutions. Second-line treatment includes probiotics and antimotility agents such as loperamide. Antibiotics (quinolones) should be reserved for severe febrile forms of the condition. PMID- 11499143 TI - [Study of vaccination for travel shows: serious gaps in polio, diphtheria and tetanus vaccination]. AB - In a retrospective study, we analyzed the immune status of 1134 travellers (621 males, 513 females), by comparing actual with recommended vaccinations, in particular with regard to malaria. 61.6% of travellers showed complete immunization against diphtheria, 73.3% against tetanus, and 66.3% against polio. Overall, women have better immunization rates than men. The best immunization rates were seen in young adults (< 20 years). The majority of travellers had destinations in Africa (35.2%), Asia (21.95) and South America (20.9%). Accordingly, vaccinations for yellow fever and hepatitis were the most commonly indicated and implemented vaccinations, and more than 50% received antimalarial chemoprophylaxis. PMID- 11499144 TI - [Screening lowers breast cancer mortality. Motivate your patients for mammography]. AB - Breast cancer is the most common malignant disease in women under 70, and the most common cause of death in those under 50. Causal prophylaxis is unknown. Only early diagnosis prior to the onset of lymphogenic or hematogenic metastasization improves the prognosis. Since the early nineteen-sixties, large-scale studies have shown that screening mammography can reduce mortality. Furthermore, early diagnosis enables breast-conserving treatment in up to 70% of cases. Disadvantages of screening mammography currently under discussion are the radiation burden, the number of false-positive diagnoses prompting biopsy and incurring costs. In Germany, the cost of screening mammography in women with no relevant history or clinical symptoms is not borne by the state-affiliated health insurance carriers. Currently, ongoing pilot projects are investigating quality controlled early diagnosis with mammography. PMID- 11499145 TI - [Systolic or diastolic blood pressure. Which value is more important?]. PMID- 11499146 TI - [When you send your patient for surgery. How risky is anesthesia?]. PMID- 11499147 TI - [Prevention of skin cancer. Sunscreens from the washing machine]. PMID- 11499148 TI - [Erectile disorders. Diagnosis and therapy]. PMID- 11499149 TI - [Occasional and spasmodic pain. Thoracic pain with normal coronary arteries]. PMID- 11499150 TI - [Modern osteoporosis therapy. Only once weekly against osteoporosis]. PMID- 11499152 TI - [Migraine headache. Severe attacks: triptans are essential]. PMID- 11499151 TI - [Exercise-induced asthma. The spray belongs in the "athletic shoe"]. PMID- 11499154 TI - [When there is a choice.... General anesthesia or regional anesthesia?]. PMID- 11499153 TI - [Therapy of type 2 diabetes. Effective control of postprandial glucose surges]. PMID- 11499155 TI - [Analgesia in obstetrics. Birth without pain?]. PMID- 11499157 TI - Exclusive or concurrent competence to make medical decisions for adolescents in the United States and United Kingdom. PMID- 11499156 TI - Is HIV a disability under the Americans with Disabilities Act: unanswered questions after Bragdon v. Abbott. PMID- 11499158 TI - Consent to sperm retrieval and insemination after death or persistent vegetative state. PMID- 11499159 TI - Defining "willful" remuneration: how Bryan v. United States affects the scienter requirement of the Medicare/Medicaid anti-kickback statute. PMID- 11499160 TI - The tobacco industry and the First Amendment: an analysis of the 1998 Master Settlement Agreement. PMID- 11499161 TI - "Calling Dr. Love": the physician-patient sexual relationship as grounds for medical malpractice--society pays while the doctor and patient play. PMID- 11499162 TI - Evolution. Bickering over old bones. PMID- 11499163 TI - Perspectives. No end in sight to rising premiums, Wall Streeters say. PMID- 11499164 TI - Local temperature changes and human skeletal muscle metabolism. AB - The aim of this review is to describe the effects induced by local temperature changes on human skeletal muscle metabolism. More specifically, we will consider the influence of temperature on the mechanical properties of muscle contraction, on aerobic metabolism, anaerobic metabolism and on the Lohmann reaction. The text has been voluntarily organized on the basis of a simple bioenergetic model describing the different energy fluxes appearing in the muscle system. This approach should better highlight some of the points that still need to be investigated. Although it was not always possible to restrict the discussion to human muscle, the references report mainly data obtained directly on humans or on isolated human fibres. A short comment on skeletal muscle temperature measurement techniques, on humans, is also included. PMID- 11499165 TI - Ergonomics of living environment for the people with special needs. AB - A safe, convenient, sound and healthy living environment is the prerequisite for a good house for the people with special needs. The intention of making a house in such a way that it solves basic problems of fixture and fittings. However the construction phase of a good house is a critical to design inside and outside structures. Often the builders do not know all the factors to be considered that can maintain a safe, hygienic and healthy environment. It is believed that when housing is ergonomically furnished, then a maximum benefit will be achieved. To meet with an individual's specific needs, an analysis of user's requirement is the most important factors to be considered in the design of special houses. Users' data such as anthropometric dimension, users' choices and preferences are also necessary to design a suitable living environment. In this regard, this paper illustrates some ergonomic features to design and develop good houses in terms of how people with restricted mobility and communication can truly be helped residing in their homes and performing their daily living activities. Users' social, medical and engineering needs are highlighted following the process of disability, ageing, or impairments to achieve the maximum level of benefits, and ensuring safe and sound living. PMID- 11499166 TI - HFs/ergonomics of assistive technology. AB - An assistive device is designed to accommodate the special needs of disability that can help people with physical, mental or cognitive challenges go through their day-to-day activities with less difficulty. An assistive device usually provide alternatives to functional limitations imposed by the client's disorder, and thereby minimising rehabilitation costs. It is therefore important to know about how assistive technology will function in all the possible aspects of such disabilities and impairements. When designing a technical device, particularly in conjunction with the target user group, ergonomic issues are therefore important to find out the extent to which an assistive device is convenient or not, and to check the quality performance of assistive technology. Since the question of the match or mismatch of an assistive device and a disabled person requires much attention, it is therefore suggested that paying attention on how an assistive device be ergonomically designed and developed is important. Ergonomic applications are to be applied for increasing motivation of prospective customers through innovative performance of AT. The authors believe that there are opportunities in ergonomic applications to manufacture an assistive device as unique, cost saving, and allows less exertation and reduces energy consumption when it is used. Hence this paper highlights human factors and/or ergonomics consideration in the process of design and development of assistive devices synchronising with gerontechnological research and development aiming to emphasise user's requirement. PMID- 11499167 TI - Thermoregulatory responses to low-intensity prolonged swimming in water at various temperatures and treadmill walking on land. AB - The purpose of the present study was to examine the effect of water temperature on the human body during low-intensity prolonged swimming. Six male college swimmers participated in this study. The experiments consisted of breast stroke swimming for 120 minutes in 23 degrees C, 28 degrees C and 33 degrees C water at a constant speed of 0.4 m.sec-1 in a swimming flume. The same subjects walked on a treadmill at a rate of approximately 50% of maximal oxygen uptake (VO2max) at the same relative intensity as the three swimming trials. Rectal temperature (Tre) in 33 degrees C water was unchanged during swimming for 120 minutes. Tre during treadmill walking increased significantly compared to the three different swimming trials. Tre, mean skin temperature (Tsk) and mean body temperature (Tb) in 23 degrees C and 28 degrees C water decreased significantly more than in both the 33 degrees C water and walking on land. VO2 during swimming in 23 degrees C water increased more than during swimming in the 28 degrees C and 33 degrees C trials; however, there were no significant differences in VO2 between the 23 degrees C swimming trial and treadmill walking. Heart rate (HR) during treadmill walking on land increased significantly compared with HR during the three swimming trials. Plasma adrenaline concentration at the end of the treadmill walking was higher than that at the end of each of the three swimming trials. Noradrenaline concentrations at the end of swimming in the 23 degrees C water and treadmill walking were higher than those during the other two swimming trials. Blood lactate concentration during swimming in 23 degrees C water was higher than that during the other two swimming trials and walking on land. These results suggest that the balance of heat loss and heat production is maintained in the warm water temperature. Therefore, a relatively warm water temperature may be desirable when prolonged swimming or other water exercise is performed at low intensity. PMID- 11499168 TI - Blood pressure and endocrine responses of healthy subjects in cold pressor test after acutely increased dietary sodium intake. AB - The objective of the study was to compare blood pressure and endocrine responses in a cold pressure test in young healthy subjects who had shown increased blood pressure during an acutely increased sodium intake. Subjects (n = 53) added 121 mmol sodium into their normal diet for one week. If the mean arterial pressure had increased by a minimum of 5 mmHg compared to the control measure, they were selected for the experiments. The selected subjects (n = 8) were given 121 mmol supplemental sodium d-1 for 14 days after which they immersed the right hand into a cold (+10 degrees C) water bath for 5 min. The blood pressure increased (P < 0.05) during the test and was independent of the sodium intake. The plasma noradrenaline increased from 2.41 +/- 0.38 nmol l-1 to 2.82 +/- 0.42 nmol l-1 (P < 0.05) with normal diet and from 1.85 +/- 0.29 nmol l-1 to 2.40 +/- 0.37 nmol l 1 (P < 0.05) with high sodium diet. The starting concentrations and the endpoint concentrations were statistically similar. The plasma levels of natriuretic peptides (NT-proANP, ANP and BNP) did not change during the test, and the concentrations were independent of the sodium diet. To conclude, acutely increased sodium intake does not change blood pressure or hormonal responses in a cold pressor test in young healthy subjects. PMID- 11499169 TI - Anatomical considerations of the ankle. AB - A complete knowledge of the complex anatomy of the ankle joint and surrounding soft tissue structures is vital for the ankle surgeon. Optimal incision site placement provides the surgeon with direct visualization of all structures that need to be inspected and avoids the vital structures that might be encountered. The surgeon with a thorough understanding of ankle anatomy can avoid possible complications associated with poor incision placement. PMID- 11499170 TI - Tarsal tunnel syndrome. AB - Most authors agree that surgical decompression is the treatment of choice for tarsal tunnel syndrome when conservative treatment fails. Overall, the results of surgical treatment for tarsal tunnel have been favorable. Studies have shown that surgical release improves or resolves symptoms of tarsal tunnel syndrome in 85% to 90% of cases. PMID- 11499171 TI - Peroneus brevis tendon tears. AB - Tears of the peroneus brevis tendon are more frequent than reported in the literature. Because of the vague pain associated with structures of the lateral ankle, peroneal tears are frequently misdiagnosed. Physical signs such as swelling along the course of the peroneal tendon sheath, pain with eversion, and subluxing tendons are diagnostic of peroneal pathologic conditions. The cause of peroneal tears is not completely understood. Possible causes include subluxing peroneal tendons, a sharp posterior ridge of the fibula, overcrowding of the peroneal groove, instability of the superior peroneal retinaculum, lateral ankle instability, contraction of the peroneus longus, hypovascularity of the peroneus brevis tendon, and a shallow peroneal groove of the fibula. Although conservative measures are almost always attempted, surgical repair of peroneus brevis tears remains the standard of care. Debridement and tubularization are recommended for less extensive tears. In more severe cases, resection of the damaged tendon and tenodesis of the proximal and distal segments to the peroneus longus are necessary. Return to maximum activity is prolonged, but with proper patient selection, evaluation, and treatment, good to excellent results can be expected. PMID- 11499172 TI - Tenodesis for chronic lateral ankle instability. AB - The goal of lateral ankle stabilization is restoration and stability without any functional deficit. Obviously, only anatomic reconstruction can prevent deficits in range of motion. As a result, motion loss is unavoidable with tenodesis procedures. Unfortunately, no procedures are available for anatomic reconstruction of subtalar joint instability. Therefore, when there is objective evidence of subtalar joint instability, tenodesis procedures must be considered. The authors believe that stability is more important than range of motion when degenerative changes are present within the ankle joint. Tenodesis results are good for the short term (less than 5 years) but may deteriorate over time (after more than 9 years). Some residual pain is common following tenodesis procedures. PMID- 11499173 TI - Evaluation and management of high ankle sprains. AB - Owing to the variability of injury, a mechanism of injury other than external rotation of the foot should not rule out a high ankle sprain. One must consider syndesmotic injury in the differential diagnosis of common ankle sprains and trauma. A high index of suspicion must lead the physician through clinical and radiographic examinations. Early and appropriate treatment of a high ankle sprain can greatly decrease a prolonged return to activity. Patients, especially competitive atheletes, must be educated regarding the morbidity of the condition. PMID- 11499174 TI - Anterior ankle abutment. AB - Anterior ankle abutment can be problematic for both athletes and for persons whose occupations require repeated loading of the anterior ankle joint. Nonoperative care is often not satisfactory for the individual who demands painfree activity. Because the presence of anterior osteophytes has been shown to represent the early stages of degenerative disease of the ankle joint, operative intervention may provide the best option to decelerate the arthritic process. Open ankle debridement or arthroscopic debridement can be used to remove the osteophytes and inspect the ankle joint. Care must be taken to examine the joint adequately to ensure that the impingement is not caused by lateral ankle instability. Failure to recognize the cause of anterior ankle impingement will result in an unsatisfactory result. When the proper diagnosis is made, surgery can result in pain-free motion that allows the individual to return to painfree activity. PMID- 11499175 TI - Ankle arthroscopy. AB - The speed of the evolution in ankle arthroscopy is staggering. Indications for the procedure continue to grow, and surgeons' skill in applying the latest technology to perform surgery better has advanced significantly in a short period of time. Certainly the biggest limitation to this point has been limited access to lesions in the posterior ankle joint. As proficiency and technology improve, better arthroscopic access will be gained to lesions that have traditionally required open arthrotomy. PMID- 11499177 TI - Mosaicplasty for the treatment of osteochondral defects of the ankle joint. AB - Autogenous osteochondral transplantation is a relatively new procedure for the treatment of osteochondral lesions of the ankle joint. The procedure is reserved for larger stage III and stage IV lesions. The corresponding author has had good preliminary results with a high level of patient satisfaction. The procedure is relatively straightforward and reliable. The need for a second surgical site has not proved to be a great disadvantage of the procedure; no complications have occurred to date at the knee donor site. This procedure, which was first described for use in the knee joint, shows excellent promise for use in the ankle and warrants larger investigational studies to assess outcomes. PMID- 11499176 TI - Osteochondral fractures of the talus. AB - Osteochondral fractures of the talus have become increasingly reported as awareness of this injury following ankle trauma has grown. Because hyaline cartilage does not have the ability to repair itself fully, surgical intervention is often necessary to create an optimal healing environment. Early recognition and appropriate diagnostic studies can aid the surgeon in producing favorable results. PMID- 11499178 TI - Posttraumatic ankle arthrosis. AB - Until there is a total ankle implant developed that stands the test of time, ankle arthrodesis will continue to be the gold standard in the operative treatment of the arthritic ankle joint. Ankle arthrodesis techniques include minimally invasive methods that can be performed via arthroscopy or with miniarthrotomy. Extensile techniques include extra-articular fusions, the transfibular approach, removal of both malleoli, and the Blair fusion. An ankle arthrodesis can be fixated utilizing either internal or external fixation. The form of fixation chosen depends on availability and surgeon preference and/or experience, and most importantly, which is the optimal method for the patient. Although ankle arthrodesis is the mainstay treatment for posttraumatic arthrosis, innovations and technical improvements have resulted in reconstructive alternatives for ankle arthritis, including distraction ankle arthroplasty, supramalleolar osteotomy, and total ankle replacement. Advances in biomaterials and instrumentation have allowed for the evolution of arthrodesis techniques as evidenced by the myriad of techniques reported in the literature. PMID- 11499179 TI - The use of cancellous screws as an alternative in plate fixation of fibular fractures. AB - Altering standard AO/ASIF techniques when performing open reduction with internal fixation of fibular fractures by using cancellous screws in place of cortical screws appears to be a viable option. By eliminating a few steps in screw application, the additive effect can lead to a significant decrease in operative and tourniquet time. Close inspection of hardware postoperatively has failed to show any signs of screw loosening or delay in fracture healing. In those cases when hardware has had to be removed, it was noted that screw tightness had not been altered. This topic has been anecdotally discussed within our institution and will be the subject of future scientific investigation. The authors feel that the application of fibular plates with cancellous screws is an acceptable alternative to standard orthopedic technique. Although we do not routinely vary from standard AO/ASIF technique, we feel this particular alteration does not affect outcome other than reducing surgical and tourniquet time, therefore reducing the surgical risks. PMID- 11499180 TI - Anterior talar dome as an alternative donor site for osteochondral transplantation for medial talar dome lesions. AB - An alternative donor site for osteochondral transplantation for medial osteochondral defects of the talus eliminates the need for a second surgical site and decreases operative time. Additionally, the donor osteochondral grafts topographical shape and contour more closely resembles the medical talar shoulder than grafts harvested from the lateral femoral condyle or the superolateral margin of the intra-articular notch. This approach has been used with good preliminary results and no evidence of donor site complications. Long-term follow up will be needed to adequately evaluate the validity of this procedure. Further investigation and clinical evaluation is warranted. PMID- 11499181 TI - Advances in the management of epithelial ovarian cancer. AB - More than 23,400 new cases of ovarian cancer and 13,900 deaths are expected in the United States this year. Epithelial ovarian cancer is the most common histologic type of ovarian malignancy. Although there have been advances in the chemotherapeutic treatment of ovarian cancer, the five year survival of women with advanced-stage disease is 25-30%. Because the disease is typically asymptomatic until the disease has metastasized and because effective screening strategies are not unavailable, 70-75% of women present with advanced-stage disease. Of ovarian cancer cases, 90-95% are sporadic and 5-10% associated with germ-line mutations, including BRCA1 and BRCA2. Known risk factors for ovarian cancer include nulliparity and a strong family history of ovarian cancer. The use of oral contraceptives is known to decrease the risk of ovarian cancer: five years of use will decrease the risk by 50%. The staging of ovarian cancer (according to the International Federation of Obstetrics and Gynecology) requires surgical exploration. Determining the extent of disease is essential to appropriate management. Survival in patients with metastatic disease is improved in those who undergo optimal primary cytoreductive surgery. Adjuvant chemotherapy is recommended in patients with high-risk, early-stage disease and all patients with advanced-stage disease. Standard chemotherapy is a combination of paclitaxel and carboplatin. Selected patients with recurrent disease can undergo secondary cytoreductive surgery. Second-line chemotherapy for patients who initially respond to paclitaxel and carboplatin and who have a prolonged disease progression-free intervals (longer than 12 months) can be re-treated with either drug or both. Those whose responses to initial therapy were less successful can be treated with other chemotherapeutic agents--e.g., liposomal doxorubicin, topotecan, etoposide, gemcitabine or taxotere. PMID- 11499182 TI - Microlaparoscopic technique for partial salpingectomy using bipolar electrocoagulation. AB - OBJECTIVE: To describe a technique of performing a partial salpingectomy using a small-diameter (2-mm) laparoscope and bipolar electrocoagulation. STUDY DESIGN: Sixty consecutive women desiring permanent sterilization underwent laparoscopic partial salpingectomy using a 2-mm transumbilical laparoscope and secondary midline sites suprapubically and midway above the pubis. A midportion of the tube was coagulated using Kleppinger forceps, transected with scissors and removed using grasping forceps. RESULTS: Additional time to remove both coagulated tubal segments averaged 4 minutes (range, 3-10). Each segment (mean, 1.5 cm; range, 0.9 2.4 cm) was confirmed in the operating room, then histologically. The transected tubal edges were separated with no thermal injury to nearby structures and with no mesosalpingeal hemorrhage. No cases required conversion from microlaparoscopy to a traditional method, and recovery time was not prolonged. The puncture sites healed well without sutures. CONCLUSION: Successful removal of electrocoagulated tubal segments with histologic confirmation was undertaken microlaparoscopically, with minimal additional operative time. PMID- 11499183 TI - Controlled ovarian hyperstimulation. Relationship of early serum E2 levels to the ultimate response of oocyte donors. AB - OBJECTIVE: To assess the predictive value of measuring serum day 5 E2 and in vitro fertilization (IVF) cycle outcome of oocyte donors undergoing controlled ovarian hyperstimulation (COH). STUDY DESIGN: Retrospective data analysis. RESULTS: Day 5 E2 significantly correlated with the peak serum E2 and number of retrieved mature oocytes. The pregnancy rates associated with stimulated day 5 E2 > 70 pg/mL were significantly greater than in cycles with levels < 70 pg/mL (58%, 71/123, versus 25%, 15/60; P < .05). CONCLUSION: The cycle performance of an oocyte donor can be predicted by measuring cycle day 5 serum E2 during COH. Poor prognosis cycles can be identified and discontinued, saving patients and the program unnecessary exspense. PMID- 11499184 TI - Oral misoprostol vs. intravaginal prostaglandin E2 for preinduction cervical ripening. A randomized trial. AB - OBJECTIVE: To compare orally administered misoprostol with intravaginal prostaglandin E2 for cervical ripening and labor induction. STUDY DESIGN: Patients presenting with medical or obstetric indications for labor induction whose Bishop's score was < or = 6 were randomly allocated to receive either 50 micrograms of oral misoprostol or 4 mg of intravaginal prostaglandin E2. If adequate cervical ripening (Bishop score of 9 or cervical dilatation of 3) or active labor did not ensue, repeat doses of each medication were administered every four hours. A maximum of six doses of either oral misoprostol or intravaginal prostaglandin E2 was permitted. Intravenous oxytocin was subsequently administered according to a standardized infusion protocol. RESULTS: Sixty patients were enrolled, with 29 randomized to the oral misoprostol arm and 31 to the prostaglandin E2 group. The data on 58 patients were eligible for analysis. Delivery occurred within 48 hours in 96.4% (27/28) of those administered oral misoprostol as compared to 76.7% (23/30) of those who received intravaginal prostaglandin E2 (P = .03). The mean time intervals from the start of induction to delivery were similar between the two groups (1,496 +/- 120 vs. 1,723 +/- 230 minutes, P = .40). No statistically significant differences existed between the two groups with respect to intrapartum complications, tachysystole, uterine hyperstimulation or adverse neonatal outcomes. CONCLUSION: Oral administration of misoprostol is an effective alternative to intravaginal prostaglandin E2 for preinduction cervical ripening. PMID- 11499185 TI - Norgestimate. From the laboratory to three clinical indications. AB - This review of preclinical studies and clinical trials of efficacy and safety examines the relation between structure and function in the norgestimate (NGM) molecule, describes the pharmacologic characteristics of NGM and evaluates clinical experience with NGM in oral contraception (OC), treatment of hyperandrogenism in women and hormonal replacement therapy (HRT). NGM is a progestin of the 19-norsteroid series with an oxime group on C-3. In women, only low serum levels of NGM can be detected for five hours after ingestion. NGM is swiftly converted into its main metabolite, the 17-deacetylated norgestimate (norelgestromin), which carries the progestogenic properties of NGM. The metabolite reaches a mean peak concentration of 3,500 pg/mL 1.5 hours after intake and has a half-life of > 24 hours. The progestogenic potency of NGM and its main metabolite is comparable to that of progesterone. The doses of NGM in OCs effectively inhibit ovulation and control uterine bleeding. In the triphasic NGM/ethinyl estradiol (EE) OC, the total monthly load of progestin is only 4.5 mg. NGM has a low androgenic impact and does not interfere with the positive metabolic actions of estrogens, notably the estrogen-induced increase in high density lipoprotein levels. OCs with NGM and EE increase the serum concentration of sex hormone binding globulin threefold, augmenting the binding of circulating testosterone and reducing free testosterone levels by 50%. Consequently, OCs with NGM are therapeutic for hyperandrogenic symptoms, such as acne. In a new type of HRT three-day dosing with 17 beta-estradiol (E2) alone is followed by three-day dosing with E2 plus NGM. This regimen treats vasomotor symptoms, protects the endometrium from hyperproliferation and is associated with a favorable lipid profile. NGM is a versatile progestin suitable for medical use from adolescence through the reproductive years to menopause. PMID- 11499186 TI - Lack of prenatal care in a traditional society. Is it an obstetric hazard? AB - OBJECTIVE: To examine the association between lack of prenatal care (fewer than three visits at any prenatal care facility) and perinatal complications in the Bedouin parturient population where lack of prenatal care is not associated with absence of prenatal services, substance abuse or marital status. STUDY DESIGN: The study population consisted of all Bedouin women lacking prenatal care (n = 7,601) who gave birth between the years 1990-1997 in the Soroka University Medical Center. The analysis consisted of a comparison of labor and delivery outcomes in women without prenatal care to outcomes in women who had some prenatal care. RESULTS: During the years 1990-1997 there were 36,281 singleton deliveries to Bedouin women at our institution. Of those, 20.9% did not receive prenatal care. Mothers in the no-prenatal-care group tended to be in the extremes of their reproductive cycles (< 18 years, > 35 years) and were of higher parity (P < .001) than those receiving prenatal care. There were more deliveries prior to 32 weeks of gestation in the no-prenatal-care group (2.8%) in comparison to the prenatal care group (1.5%, P < .001) and fewer postterm deliveries (> 42 weeks, P < .01). The incidence of low birth weight (< 2,500 g) in the no-prenatal care group was higher than in the prenatal-care group (11.2% vs. 8.4%, P < .001). Women who did not receive adequate prenatal care had statistically significantly higher rates of antepartum fetal death (OR = 1.8, 95% CI 1.4-2.3, P < .001), intrapartum fetal death (OR = 2.38, 95% CI 1.2-4.5, P < .03) and postpartum fetal death (OR = 1.60, 95% CI 1.2-2.1, P < .001). Multiple logistic regression models were used to analyze the independent contribution of lack of prenatal care to perinatal mortality and to very-low-birth-weight newborns. In both models lack of prenatal care was an independent contributing factor. CONCLUSION: Lack of prenatal care is an independent contributor to perinatal mortality and low birth weight in a traditional society. In light of the high percentage of lack of prenatal care in Bedouin society, special attempts should be made to encourage women to use the available prenatal services. PMID- 11499187 TI - Effect of feedback on obstetrics and gynecology residents' teaching performance and attitudes. AB - OBJECTIVE: To evaluate obstetrics and gynecology residents' teaching performance, perception of the importance of teaching and satisfaction with their evaluations after the institution of an oral or written medical student feedback and award system. STUDY DESIGN: Residents at a single, university-based obstetrics and gynecology program were assigned to receive either oral, written or no medical student feedback on their teaching skills in a prospective, randomized, controlled trial. Students rated resident performance in seven teaching categories. Residents' scores per six-week block were evaluated for one year. Questionnaires addressing resident attitudes toward feedback were collected at baseline and at 6 and 12 months. All residents then received written feedback and public awards for high scores for an additional year. RESULTS: After 12 months of feedback there were trends toward improvement in several of the teaching categories and overall evaluations. None of the controls, 29% of residents receiving oral feedback and 50% of residents receiving written feedback rated teaching as more important than before. None of the controls, 57% of those receiving oral feedback (P = NS) and 88% of those receiving written feedback (P = .009), for a total of 73% of residents receiving any feedback (P = .001), thought that the amount of feedback was adequate at 12 months. Follow-up of 15 residents after one year of written feedback with an award for high evaluations showed that 60% improved their overall scores. The mean overall group score improved. CONCLUSION: A feedback and award system can lead to improved resident teaching performance as well as enhanced perception of residents' role as teachers and greater resident satisfaction. PMID- 11499188 TI - Video vs. CD-ROM for teaching pelvic anatomy to third-year medical students. A comparison. AB - OBJECTIVE: To compare the effectiveness and student acceptance of video and CD ROM review of pelvic anatomy in third-year obstetrics and gynecology students. STUDY DESIGN: A 34-item pretest was administered on the first day of the clerkship. Students were then randomized to watch a 45-minute video or use an interactive CD-ROM at home. All students took a posttest identical to the pretest. Examination scores were compared by paired and unpaired t test. Students completed a questionnaire on each method's ease, appropriateness, organization, appeal and time spent and whether they would recommend the method to others. RESULTS: One hundred seventy-six students were randomized, 88 to CD and 88 to video. There was 100% compliance, although not all questionnaire items were completed. The mean pretest score for both groups was 50%. The posttest CD mean score was significantly different from the posttest video mean (64% vs. 70%). Both groups significantly improved after the intervention. The video group noted ease of use, but only 49% recommended it for the future. The CD-ROM was recommended by 80%. Student comments favored the CD-ROM. CONCLUSION: Use of video review resulted in short-term significantly higher test scores, but students preferred an interactive CD-ROM. This method should be made available to all students during the rotation. PMID- 11499189 TI - Androgen suppression and clinical improvement with dopamine agonists in hyperandrogenic-hyperprolactinemic women. AB - OBJECTIVE: To examine the effect of dopamine agonist (DA) treatment on clinical and biochemical features in hirsute, hyperprolactinemic (HPRL) women and the relationship between prolactin (PRL) and androgens. STUDY DESIGN: We evaluated 80 hirsute HPRL women (age, 27 +/- 1 years [mean +/- SE]) with neuroleptic treatment, prolactinoma and idiopathic HPRL (12, 13 and 55, respectively). DA, mainly bromocriptine, was administered for 11 +/- 1 months. Response indicators were Ferriman-Gallwey hirsutism (FGS) and Leeds acne (LAS) scores, circulating PRL, dehydroepiandrosterone sulfate (DHEAS), free and total testosterone, and androstenedione. RESULTS: Baseline PRL correlated positively with DHEAS (r = .23, P = .03) and free testosterone (r = .36, P < .001). In all women, FGS, LAS, PRL, free testosterone, DHEAS and androstenedione decreased by 40-85% during DA treatment (P < .001). The decline in free testosterone was higher when PRL was > or = 65 ng/mL than when PRL was < 65 (P = .03) and correlated positively with basal DHEAS (r = .40, P < .001). CONCLUSION: Our data suggest a modulation by PRL of adrenal androgen production. DA treatment reduces PRL and serum androgens. It results in a significant clinical improvement in acne and hirsutism. Therefore, DA is recommended as monotherapy for hyperandrogenic. PMID- 11499190 TI - Relationship between normal amniotic fluid index and birth weight in term patients presenting for labor. AB - OBJECTIVE: To investigate whether a relationship between birth weight (BW) and amniotic fluid index (AFI) existed for term patients with intact membranes and, if so, whether it could predict macrosomia and intrauterine growth retardation. STUDY DESIGN: Two hundred thirty-one eligible women at term (37-42 weeks' gestation) had an AFI between 5.0 and 24.0 cm at the time of admission. Demographic information was collected, including BW. Statistical comparisons were made, with significance set at P < .05. RESULTS: The mean gestational age was 39.5 weeks, and the mean AFI was 11.3. There were 28 infants with BWs > 4,000 g (macrosomia). The cesarean section rate was 16.9%. Macrosomia was associated with a higher cesarean section rate (39.2% vs. 13.8%, P < .002). The mean AFI was higher for those with macrosomia (13.5 vs. 11.0, P < .001). Patients with an AFI > 15.0 had over twice the incidence of macrosomia (RR 2.72, 1.1-6.6, 32.1% vs. 14.8%, P < .027). The risk was even greater for them with an AFI > 18.0 (RR 6.4, 2.2-18.7, P < .002). A linear relationship was observed between AFI and BW (r2 .061, P < .0001). BW increased with increasing AFI. No relationship between AFI of 5-8 and BW < 2,500 g was found. CONCLUSION: Increasing AFI correlates linearly with increasing BW, and macrosomia is associated with an increased incidence of cesarean delivery. AFI > 15 carries over double the risk of birth of a macrosomic infant, while AFI > 18 has a risk of over six times. No association between low normal AFI and a growth-restricted infant was found. PMID- 11499191 TI - Arterial occlusion after surgery for ovarian malignancy. A case report. AB - BACKGROUND: Inherent in the risks of pelvic surgery is the postoperative morbidity and mortality associated with the procedure. The gynecologic oncology patient presents further risks for several reasons, including a relative state of hypercoagulability. Anticipation of the problem and early recognition are the keys to decreasing the long-term sequelae of a complication. A well-recognized complication of gynecologic oncology surgery is venous thromboembolism. Advances allow the prophylaxis, recognition and treatment of this problem. Acute arterial occlusion is a much less common but potentially devastating complication. CASE: A woman underwent surgery for ovarian malignancy and developed bilateral arterial occlusion of the lower extremities postoperatively. CONCLUSION: Understanding the various genetic defects that predispose certain individuals to hypercoagulability will help identify patients with a particularly high risk of developing acute arterial occlusive disease. PMID- 11499192 TI - Third consecutive complete posterior uterine rupture. A case report. AB - BACKGROUND: Management of pregnancy in a woman who has had a ruptured uterus on more than one occasion presents a great clinical and ethical challenge to the obstetrician. CASE: This appears to be the first report on complete uterine ruptures in three consecutive pregnancies. CONCLUSION: Prolonged hospital admission, intensive antenatal surveillance, antenatal steroid administration and elective premature delivery may give the best chance for a good outcome in these pregnancies. However, despite all these efforts, there is still a high risk of an unfavorable outcome. PMID- 11499194 TI - Fetal abdominal circumference and macrosomia. PMID- 11499193 TI - Prenatal diagnosis of meconium peritonitis in twin pregnancy after intracytoplasmic sperm injection. PMID- 11499195 TI - When patient can't get to ER: what role should security officers play? AB - If someone in a hospital parking lot needs immediate medical attention, should hospital staff go out and treat the patient or call an ambulance to bring the patient inside? The answer may depend on whom you talk to. The issue drew national attention in 1998 following the death of a 15-year-old boy in Chicago who was shot by gang members. The boy's friends left him just outside the emergency room of a nearby hospital. Hospital workers refused to go out and get him, so police called an ambulance to bring him inside. PMID- 11499196 TI - Suicide prevention: what hospital security directors should know. PMID- 11499197 TI - 'Non-deadly force training' for hospital security officers. AB - Some 950 law enforcement agencies worldwide and 27 hospitals, the majority located in New York and New Jersey, are providing their police and security staffs with non-deadly force training techniques to effectively defuse, control, or disarm an attacker and to help reduce liability for everyone involved. The system was developed by CDT Training Inc., Elmwood Park, NJ. According to Thomas J. Patire, a security management consultant and president/senior instructor of CDT Training Inc., "the CDT system was originally designed for personal protection agents like bodyguards and police departments. It was expanded to other sectors because of the overall problem that society faces--inconsistent or lack of realistic, proficient non-deadly force training that can reduce liability for actions taken. Hospitals, for example, face threats daily against various staff members and members of the medical staff due to their work in various areas and contacts with patients and other hospital visitors. PMID- 11499198 TI - Tips on thwarting thieves who prey on hospital employees. AB - Following a series of hospital thefts in New Jersey, the state's IAHSS Chapter, along with the New Jersey Hospital Association and the Englewood, NJ Police Department, held a seminar on "Preventing Hospital Thefts by Outsiders: Establishing a Dialogue and Strategy with Local Police." The featured speaker at the conference was Detective Timothy Torell, Englewood Police Department. The department solved a number of hospital thefts in Englewood that were perpetrated for years by one individual. In attendance were 80 law enforcement executives and hospital security directors, who learned more about. PMID- 11499199 TI - Hospitals and disasters: how they fared; what they learned. AB - Disasters happen, be they nature-driven like a tornado, hurricane, or ice storm; accidents like plane crashes or train wrecks; or failures or defects in critical power systems. When they do take place and your hospital is called on to respond, a well-thought-out disaster plan, augmented by regular drills, can save the day. But, as we have reported in our coverage of many major disasters over the years, no plan covers all eventualities and there is always some unforeseen challenge that has to be overcome and incorporated into future plans. In this report, we'll give details of how hospitals and their security officials responded to an external disaster and an internal situation that was potentially disastrous, and what these real-life events taught them. PMID- 11499200 TI - Case of the month. Awkward assessments. PMID- 11499201 TI - Olympic trials. PMID- 11499202 TI - Extrication challenges. Cement mixer entrapment. PMID- 11499203 TI - Extrication challenges. 10 tips for EMS crews working at extrication scenes. EMS on scene. PMID- 11499204 TI - Extrication challenges. Under the metal. PMID- 11499206 TI - Get the data. AB - Numerous studies affirm the benefit of rapid defibrillation with AEDs. Most of the data have been acquired in emergency medical response systems with EMTs or firefighters providing defibrillation. Yet interest is growing in the provision of rapid defibrillation by other responders, including security officers, flight attendants and laypersons trained as targeted responders. This expansion is embraced in community early defibrillation. A multicenter, prospective, randomized, controlled trial was recently launched to define the benefit of community early defibrillation. Data from this trial will likely identify the most effective deployment of AEDs, frequency and appropriateness of use, time of defibrillation before EMS arrival, device maintenance issues and--most importantly--survival benefit when compared with control groups without AEDs. PMID- 11499205 TI - How to launch a community early defibrillation program. AB - More than 60 emergency services professionals attended an EMS Today 2001 preconference symposium, "How to Champion Public Access Defibrillation in Your Community: Leadership, Art and Science," hosted by AED manufacturer Philips/Heartstream. Their goal: to learn the practicalities of starting and maintaining an early defibrillation program in their communities. The recent increase in research and attention to community early defibrillation programs led Philips/Heartstream to host the symposium, which featured cardiac arrest survivor Diane Jackson, as well as three community early defibrillation program advocates who've implemented early defibrillation programs in their communities. Their advice for kick-starting early defibrillation programs in your community follows. PMID- 11499207 TI - Heart-to-heart talk. Roundtable discussion. AB - This past August the American Heart Association (AHA) released a major revision of its recommendations for emergency cardiovascular care (ECC). The 2000 guidelines simplify CPR techniques for the layperson and strongly promote the use of community early defibrillation. What repercussions can we expect from these changes? We asked five experts to join a roundtable discussion of the BLS aspects of these new guidelines. Their conversation follows. PMID- 11499208 TI - State legislation to support family caregivers. Opportunities for partnerships abound. AB - This article focuses on three examples of state legislation that can improve the quality of life of family caregivers while enhancing the role of home care agencies. PMID- 11499209 TI - New directions in home & community-based services at the state level. PMID- 11499210 TI - Moving toward community-based care. Vermont's 10-year odyssey. AB - In Vermont, too much was spent on institutional care and not enough on home and community-based services. One study, written for Vermont's General Assembly, suggested that Vermont should work toward a gradual shift in spending with more spent on home care services and less on nursing home care. PMID- 11499211 TI - Implementing the Olmstead decision. Are states moving forward or treading water? PMID- 11499212 TI - The evolution of assisted living. A view from the states. AB - Use of nursing homes has declined significantly with the expansion of home care services and the explosion of assisted living facilities. To give consumers more choices, many states allow assisted living facilities to provide higher levels of service that enable residents to continue to live as independently as possible as their needs change. PMID- 11499213 TI - New Mexico breaks the Medicaid mold. AB - On September 1, 1999, the New Mexico Medical Assistance Division implemented the Medicaid Personal Care service. The goal of Medicaid Personal Care is to improve the quality of life for the disabled and the elderly, and prevent them from having to enter a nursing facility. Personal Care allows consumers to live in their own homes and achieve the highest level of independence they can reach. PMID- 11499214 TI - Expanding roles: delegating tasks to home care aides. PMID- 11499215 TI - HCU's online research resource. AB - Home care providers should be active consumers of research as well as participants in collaborative research efforts. Knowledge derived from good research is essential to their success in negotiating the complex environment and delivering the care their patients deserve. PMID- 11499216 TI - Hip fracture research results related to home care. AB - Hip fracture is a topic we are likely to hear more about as health care professionals continue to work collaboratively to prevent increases in incidence, cost, and poor outcomes associated with hip fracture. Research shows that home care is the optimum place for these patients and agencies should continue to strive to improve outcomes and the consistency of care across settings. PMID- 11499217 TI - The dance of federalism: power to the people. PMID- 11499218 TI - The heart of Texas home care. AB - It is only fitting that a state as large and ethnically diverse as Texas should have such a broad array of home care programs delivered through licensed home and community support services agencies. Over the past 20 years, Texas has moved from a one-third community/two-thirds nursing home mix of long-term care clients to the exact opposite. The personal care programs have provided a stable base for expansion into the waiver programs. PMID- 11499219 TI - China's lifestyle choice. PMID- 11499220 TI - Full-court cleanup. PMID- 11499221 TI - Power struggle. Who's in charge here?. PMID- 11499222 TI - Canine candy stripers. PMID- 11499223 TI - A little help from your friends. PMID- 11499224 TI - To DEET or not to DEET? PMID- 11499225 TI - Senate subcommittee to probe GPO influence. PMID- 11499226 TI - Price survey. No hike in sight for glove prices. PMID- 11499227 TI - Santa Clara auditors verify GPO savings. PMID- 11499228 TI - [Methods of identification and localization of ethmoid and sphenoid osteomeningeal breaches]. AB - The accurate localization of cerebrospinal fluid (CSF) fistula presenting as rhinorrhea is an essential requirement for successful dural repair, in order to eliminate unsuccessful surgical exploration, and to achieve a definitive closure of the osteo-dural defect. The authors report their experience in 22 patients, and propose a simplified attitude: realization of high resolution CT scan in all cases, in first choice, and a MRI with MR cysternography sequences, in second choice, in case of negative CT scan. PMID- 11499229 TI - [Ethmoid adenocarcinomas: retrospective study of 76 patients]. AB - From the retrospective study of 76 adenocarcinomas of the ethmoid sinuses, results were expressed in two terms: the morbidity related to surgery and the oncologic outcome. This case study extends between 1975 and 2000. It includes 71 men and 5 women, with an average of 61 years. 81% of them works in wood dust, with a mean duration of exposure of 26 years. The diagnosis of adenocarcinoma is realized in the three months after the first signs, essentially rhinologicals, next neurologicals and ophthalmologicals. The majority of tumors were classified as T3N0M0, i.e. 57.89%. The treatment of this tumors is surgical: 34.2% surgical only and 59.2% with radiotherapy. The transfacial approach (paralateronasal and degloving) and the combined surgery are respectively performed in 36 patients and 35 patients. 23.2% patients had a local recurrence, and 10% developed cervical nodes and systemic metastasis. Survival rate based on the Kaplan-Meier actuarial method is 82% at 3 years, 80% at 5 years, 72% at 10 years. The prognosis of ethmoidal cancer is strictly correlated to local control. Local recurrence is statistically more likely in patients with involvement of the dura, brain and sphenoid sinus. With the analysis of the carcinologics results, we discuss the therapeutics indications of the adenocarcinomas and a new classification. Taking in account the involvement of the dura, sphenoid and orbit. We did not find any statistical differences between T3 patients treated by combined approach (n = 13) or by transfacial approach (n = 15). PMID- 11499230 TI - Fungal infections of the paranasal sinuses. AB - Fungal infections can be mainly grouped into four types. The invasive forms are acute sinusitis (fulminant), chronic sinusitis (indolent), whereas the non invasive forms are mycetoma and allergic fungal sinusitis. From December 1993 to December 1997, 27 cases of fungal sinusitis, 22 of which were noninvasive forms, and 5 of which were invasive forms, were treated and are presented in this study. When we classified the patients with fungal sinusitis, 11 were diagnosed as mycetoma, 9 as allergic fungal sinusitis, 3 as acute fulminant sinusitis and 2 as chronic indolent sinusitis, while 2 patients were not included in our four groups of sinusitis. In all mycetoma cases the active agent was Aspergillus. Patients with non invasive forms of sinusitis were all treated with endoscopic sinus surgery. 2 of the patients with invasive forms of sinusitis underwent maxillectomy and they were given Amphotericin-B. With a mean follow up of 20 months, only 3 recurrences were seen. The infection recurred in 2 patients with allergic fungal sinusitis and 1 patient with chronic invasive sinusitis. However, 2 patients with acute fulminant invasive sinusitis died before they were operated on, and 1 patient died postoperatively. PMID- 11499231 TI - Complicated fungal sinusitis. Clinical and therapeutic aspects. AB - Fungal infections of the sinuses are becoming more common and increasing number of cases are being reported in immunocompetent patients. Unfortunately a significant number of cases present with a complication. In this review of 140 patients with fungal sinusitis, 16 (11.4%) presented with an extrasinus, orbital or intracranial complication. The different fungal types, presentation and therapeutic approaches are discussed. This emphasizes the need for vigilance in dealing with patients with atypical sinus disease and to suspect fungal infections pre- and intraoperatively. Combined surgical and long term medical treatments are usually needed to prevent recurrences and spread of the infection. PMID- 11499232 TI - [Olfaction disorders: preliminary findings and results in 45 cases]. AB - In this work we present the preliminary results of a retrospective study on 45 cases of disordered sense of smell. Our approach is based on a diagnostic tripod- a thorough history (with a questionnaire for evaluation), clinical examination (nasal endoscopy), and imaging. The mean age of our patients was 42.6 years, and 60% were male. In the aetiological factors there was a predominance of naso-sinus disease (62.2%), especially inflammatory (polyposis of the nose and sinuses), while general causes (metabolic and drug-induced) accounted for 24.4%, and neurological causes a further 13.3% of cases. Treatment resulted in a favourable outcome (with partial or total recovery) in 71.1% of cases, and failed in 26.6%. PMID- 11499233 TI - [Value of endonasal endoscopic surgery in the treatment of sphenoid cerebrospinal rhinorrheas. 15 cases]. AB - Surgical management of cerebrospinal fluid leaks (CSFL) has improved these few past years with the development of paranasal sinus surgery under optical guidance. CSFL localized in the sphenoid sinus represent only 5 to 15% of all CSFL. The authors have analyzed a serie of 15 patients having undergone surgery from 1992 to 1999 for CSFL: 10 cases followed pituitary gland surgery realized through sublabial-transsphenoidal approach, 2 cases followed head trauma, 1 case followed a secondary neurosurgical procedure following recurrence of a rathke's cleft cyst, 1 case followed medical treatment of a bulky pituitary gland adenoma and 1 case was associated to an empty sella syndrome. The surgical procedure has relied on sphenoid sinus approach through the sphenoethmoidal recess (SER) (n = 8), transethmoidal approach (TE) (n = 2) or transseptal approach (TS) (n = 7) followed by a filling of the defect by a graft of fat with biological glue (n = 17). Sometimes it was associated to a cartilagenous graft (n = 7) when there was a bone defect. The average period of hospitalisation was 5.2 days (4-21), mean follow-up was 2.9 years (6 months-7 years). RESULTS: The success rate was 80% (3 failures were observed), 2 patients underwent secondary procedures by TS approach with success. Our results combined with the literature data led us to propose an alternative surgical strategy adapted to each case. Endonasal surgery under optic guidance compared to the transrhinoseptal approach represents an attractive alternative with less undesirable iatrogenic consequences. PMID- 11499234 TI - [Diagnostic value of needle biopsy and frozen section histological examination in the surgery of primary parotid tumors]. AB - The necessity of fine-needle aspiration biopsy (FNAB) in the diagnosis and treatment of parotid gland lesions is still controversial. We examined the accuracy of cytology and histology in a review of 128 parotid gland tumors who underwent surgery with FNAB, n = 102 and/or frozen section examination (FS), n = 94. The diagnostic sensibility and specificity for malignant or benign lesions was respectively 81.5% and 97.5% for FNAB and 75% and 100% for FS as compared with definite histology (110 tumors were benign and 18 malignant). Insufficient material for FNAB evaluation was found in 12 patients mainly with small tumors (p = 0.043) or with tumors located in the deep process of the parotid gland (p = 0.029). Surgery was inappropriate (superficial lobe resection for malignant tumor) because of 4 false negative FS diagnoses. FNAB offers valuable information in the diagnosis of nonsurgical lesions and permits to avoid FS if FNAB identify a benign lesion. FS remains mandatory if FNAB evaluation is not possible or suggests a neoplastic tumor. PMID- 11499235 TI - [An original case of laryngomucocele after Tucker surgery]. AB - Although discovered since nearly 2 centuries, the cystic tumours of the laryngeal ventricle remain very badly known by otorhinolaryngologists. Air-containing or cystic, these lesions can be congenital or acquired. We present an original case of laryngomucocele, acquired after a partial laryngectomy of Tucker type for a laryngeal neoplasty. Through a review of the literature, we expose in the second time, the etiopathogenic mechanisms of these lesions. If this pathology contributes to mislay the diagnosis during the post-operative survey, small surgical details at the time of the initial intervention, can prevent the appearance of this lesion. The treatment is a surgical procedure, with an endoscopic way or a cervicotomy. PMID- 11499236 TI - [Cervical and pharyngeal inflammatory pseudo-tumors, a report of two cases: clinical course and treatment]. AB - The authors report two new cases of inflammatory pseudotumours, sited in the pharynx and neck--sites in which no previous cases have so far been reported in the literature. The aetiology and clinical course of these rare benign tumours remain little understood even today. Their treatment is also not clearly described. This is most often surgical, although medical treatment is often very effective, and is sometimes appropriate. The authors begin by describing these two clinical cases, and then use the literature to give an account of the clinical course and treatment of inflammatory pseudotumours. PMID- 11499237 TI - [Hygiene and sterilization in the medical office: quality control]. PMID- 11499238 TI - The Federation is back. For-profits' lobby gets a shot in the arm from members' good year. PMID- 11499239 TI - As efficient as they have to be. Study finds mergers' cost, price savings depend on market competition. PMID- 11499240 TI - Monopoly rules change. Health system neighbors unite to share hospital. PMID- 11499241 TI - Modern Healthcare celebrates the 25th anniversary of the magazine's ownership by Crain Communications. AB - In many ways it's been a quarter-century of change. But in some ways it's been just the opposite. To mark Modern Healthcare's silver anniversary under its current ownership, the magazine's staff took a trip through time, looking at pivotal events in healthcare during the past 25 years and the people who helped shape the industry. PMID- 11499242 TI - Study questions safety standards. PMID- 11499243 TI - Judge hits secrecy of PRO probes. PMID- 11499244 TI - S.C. docs sue feds on privacy regs. PMID- 11499245 TI - Not so bad? IG: Medicare outpatient losses may be overstated. PMID- 11499246 TI - Medicine. Dying for science. PMID- 11499247 TI - [Risk factors in hypertension: a disease with fatal consequences]. PMID- 11499248 TI - [Do cells sense oxygen?]. PMID- 11499249 TI - [A new path to flu treatment?]. PMID- 11499250 TI - [St. John's wort is ineffective for severe depression]. PMID- 11499251 TI - [Oral polio vaccine/AIDS theory refuted!]. PMID- 11499252 TI - [Signal transduction of PrPc]. PMID- 11499253 TI - [The race for a new target. The human AT1 receptor]. PMID- 11499254 TI - [Pharmacologic basics: the effect of AT1 receptor antagonists]. PMID- 11499255 TI - [Pharmacologic basics: the effect of AT1 receptor antagonists]. PMID- 11499256 TI - [Concurrence to ACE inhibitors? AT1 receptor blockers and hypertension]. PMID- 11499257 TI - [Can Sartane help a failing heart? AT1-receptor antagonists in cardiac insufficiency]. PMID- 11499258 TI - [Application to the internist practice. The place of AT1 receptor antagonists]. PMID- 11499260 TI - [Measurement precision and correctness!? Blood pressure measuring instrument critical assessing]. PMID- 11499259 TI - [Recommendations regarding AT1 receptor antagonists. Selection criteria of the drug commission]. PMID- 11499261 TI - [Epidemiology and costs of bronchial asthma and chronic bronchitis in Germany]. AB - BACKGROUND AND OBJECTIVE: Asthma bronchiale (AB) and chronic bronchitis (CB) are common chronic disorders with high rates of prevalence. We performed a cost of illness study that aimed to assess the economic burden of these disorders in Germany. PATIENTS AND METHODS: Costs were estimated in 1996. In a retrospective analysis we calculated direct and indirect costs based on secondary data from governmental institutions as well as from the pharmaceutical industry. To the best of our knowledge, this is the first study which uses data for the Federal Republic of Germany following its reunification. RESULTS: Total estimated costs were DM 5.81 billion related to AB, and DM 20.17 billion related to CB. We did not include outpatient physician services in the calculation of direct costs due to a lack of adequate data. Therefore, direct costs represented only 33% and 22% of total estimated costs, respectively. The most important cost driver of direct costs were outpatient prescribed medicines, followed by hospitalization. Outpatient prescribed medicines accounted for 55% and 63% of the direct costs, respectively. Of the indirect costs of AB 43% were associated with early retirement. The largest single cost driver of indirect costs due to CB was by far loss of work, amounting to 75%. CONCLUSIONS: The data suggest that therapeutic progress and cessation of smoking can provide distinctive savings of direct costs and even more of indirect costs of AB and CB. PMID- 11499262 TI - [Fatal brain edema after ingestion of ecstasy and benzylpiperazine]. AB - HISTORY AND ADMISSION FINDINGS: A 23-year-old woman was hospitalized with headache, malaise and somnolence 11 hours after ingestion of A2 (benzylpiperazine), 7 hours after ingestion of ecstasy (MDMA), and large volume of fluids. On admission she had bradycardia (heart rate 48/min), hypertension (blood pressure 154/95 mm Hg), and reduced consciousness with diminished tendon reflexes and non-reacting pupils (Glasgow Coma Score 6). INVESTIGATIONS: Serum sodium was markedly decreased (115 mmol/l [normal 135-145]) with low plasma osmolality (246 mosm/kg [normal 280-300]). Other laboratory findings were within normal limits. TREATMENT AND COURSE: The patient had severe hypervolaemic hypotonic hyponatraemia. 40 minutes after admission she seized twice and was intubated. Brain CT scan showed massive cerebral oedema with beginning tonsillar herniation. Serum sodium concentration returned to normal within 38 hours, but the patient deteriorated neurologically with increasing tonsillar herniation detected in a second brain CT scan. The patient died 57 hours after admission. CONCLUSION: 13 cases of MDMA-associated severe hyponatraemia are reported. Intake of fluids after MDMA ingestion may lead to potentially fatal hypervolaemic hypotonic hyponatraemia with cerebral oedema. Symptoms appear about 8 hours (range 4-18) after MDMA ingestion. Even low doses of MDMA and fluids may lead to a serious outcome. The only risk factor is female gender. Measurement of serum sodium and brain CT scan is recommended in all patients with altered mental status after MDMA consumption. PMID- 11499263 TI - [Ciguatera: clinical relevance of a marine neurotoxin]. AB - HISTORY AND CLINICAL FINDINGS: A 45-year-old woman complained of gastrointestinal symptoms followed by paraesthesiae of the mouth, face, hands, feet, and trunk after she had eaten fish while she was on a journey to the red sea. Additionally, she suffered from headaches and muscle aches, weakness of the arms and legs, instability of gait as well as troubles of vision, taste, speech, and swallowing. Neurological examination revealed a cerebellar syndromE with mild dysarthria, multifocal dysaesthesiae, paresis of the arms and legs, generalized hyporeflexia, bilaterally reduced vision, and ataxic gait. INVESTIGATIONS: Laboratory work-up provided no diagnostic clues. Cranial and spinal magnetic resonance imaging were normal. The cerebrospinal fluid showed a normal cell count and a moderately impaired barrier function. Electrophysiological evaluation suggested a sensory and motor axonal neuropathy. DIAGNOSIS, TREATMENT AND COURSE: Based on the history and the characteristic clinical findings we made a diagnosis of ciguatera, a marine food poisoning caused by the ingestion of fish contaminated with ciguatoxin. Symptomatic treatment with mannitol, amitriptyline, carbamazepine, and corticosteroids started 3 weeks after the onset of symptoms proved inefficient, however, during the 21 months of follow-up there was a marked spontaneous clinical and electrophysiological reversal of symptoms. CONCLUSIONS: In the age of growing foreign tourism marine food intoxications have to be considered in the differential diagnosis of the frequently occurring travellers diarrhea. The diagnosis of ciguatera is based on the typical history of a fishmeal followed by characteristic clinical findings with predominant neurological symptoms, often including dysaesthesiae, besides gastrointestinal disturbances. In the early phase of the disease intravenous therapy with mannitol has emerged as an efficient treatment. PMID- 11499264 TI - [Cyclophosphamide]. PMID- 11499265 TI - [Approved monoclonal antibodies for in vivo diagnosis]. PMID- 11499266 TI - [Acute delirium in elderly patients--a neglected interdisciplinary problem]. PMID- 11499267 TI - [Argon laser to treat polyps]. PMID- 11499268 TI - [ACE inhibitors for therapy of cardiac and kidney insufficiency]. PMID- 11499269 TI - [Hemorrhoids during pregnancy]. PMID- 11499270 TI - [Oncologic mistletoe therapy--physicians' use and estimation of efficiency]. PMID- 11499271 TI - [Spontaneous rupture of central venous catheter]. PMID- 11499272 TI - Discounts on your Rx drugs. PMID- 11499273 TI - Fighting the HMO meanies. PMID- 11499274 TI - Trials--and errors. PMID- 11499275 TI - My life as a guinea pig. PMID- 11499276 TI - The road to rave. PMID- 11499277 TI - What happens to a dream deferred? The folly of delaying HIPAA. PMID- 11499278 TI - It's more than the technology: an interview with CIO Tim Zoph. PMID- 11499279 TI - Creating sustainable physician-IDS relationships. AB - IDSs that have faced financial difficulties as a result of owning physician practices need to assess whether employing physicians remains the best way to maintain effective relationships with the physicians. IDSs should evaluate each owned physician practice individually, asking a series of specific questions. First the IDSs should assess the value of the physician practice to the organization to determine whether a future relationship is desirable. If so, the IDS should determine whether the relationship needs to be formally established. If the relationship is likely to remain productive whether or not it is formal, the IDS should consider divesting the practice. If, however, a formal business relationship is required, the IDS needs to decide whether employment is preferable to an alternative approach, such as involving physicians in joint ventures or equity relationships. PMID- 11499280 TI - Medicare+Choice risk adjustment: don't leave money on the table. AB - The Principal In-Patient Diagnostic Cost Group (PIP-DCG) risk-adjustment methodology for Medicare+Choice organizations uses inpatient hospital encounter data to trigger payment for certain diagnoses. Improved financial performance under PIP-DCG risk adjustment for Medicare+Choice organizations and their hospital partners lies in more complete and accurate coding and better record keeping at the hospital level. It is important to track members with chronic or terminal conditions that may qualify for higher payments and to appropriately code these conditions to optimize funds that may be owed to the organization. PMID- 11499281 TI - Developing mechanisms for reporting compliance violations. AB - Both the Federal Sentencing Guidelines and the Office of Inspector General's (OIG) Compliance Program Guidance for Hospitals can assist healthcare organizations in establishing a reporting system that encourages employees to report wrongdoing. One mechanism that is commonly used is the telephone hot line. Other mechanisms that can be used in conjunction with a hot line include a drop box or post office box, written or oral reports to supervisors, an open-door policy on the part of compliance personnel, and employee exit interviews. However the reporting system is set up, it should ensure confidentiality and a policy of nonretaliation to encourage the participation of all employees. Having a sound reporting system in place will enable healthcare organizations to investigate any alleged instances of noncompliance and take corrective action before the Federal government becomes involved. PMID- 11499282 TI - AICPA standard can help improve audit committee performance. AB - The AICPA's Auditing Standards Board issued Statement on Auditing Standards (SAS) No. 90, Improving the Effectiveness of Corporate Audit Committees, in response to a related Securities and Exchange Commission (SEC) rule change. Under the standard, audit committees of publicly traded organizations must enter into discussions with their organization's auditors regarding the quality and the acceptability of the accounting principles that have been applied in the organization's financial statements. Although SAS No. 90 is intended for publicly traded entities, the standard has significant implications for all healthcare organizations in defining what constitutes an effective audit committee. To comply with the standard, audit committees should have independent directors who are rotated on and off the committee, training for committee members on the effective performance of their duties, a charter specifying committee responsibilities and membership requirements, meetings at least four times annually, and responsibility for retaining services of CPA firms to conduct the organization's annual audits. PMID- 11499283 TI - Avoiding HIPAA hype: preparing for HIPAA affordably. AB - In the scramble to prepare their organizations for HIPAA compliance, healthcare financial managers may believe they must commit significant funds to overhauling information systems and hiring consulting firms. But security and confidentiality policies required by HIPAA cannot be implemented solely by upgrading computer systems. HIPAA compliance requires a wholesale change in staff attitudes and behavior regarding patient security and confidentiality. Healthcare organizations can prepare to comply with HIPAA regulations practically and affordably by combining in-house expertise with limited assistance from outside consulting firms. An added benefit of emphasizing the use of in-house resources to prepare the organization for HIPAA compliance is that the organization can develop and retain its own experts on HIPAA. PMID- 11499284 TI - Upgrading the service mix in primary care practices may increase revenue. AB - Hospitals and health systems that own primary care physician networks want to optimize revenues when possible while still providing high-quality patient care. One way to accomplish this objective is to enhance the service mix for patients seen by the primary care practices. An analysis of the diagnosis and service mix of three primary care networks showed that the primary care practices mainly are involved with common acute and chronic conditions, which are associated with office visits that are paid at low- and mid-level rates. A comparison of the primary care networks' diagnosis and service mix with Medicare data on the mix of office visits for family practitioners, internists, and cardiologists showed, for example, that primary care physicians have more level 2 office visits and fewer level 4 office visits than cardiologists. By upgrading the patient office visits from the routine acute and chronic conditions associated with lower-level visits to more serious conditions associated with higher-level visits, primary care practices may increase total practice revenue. PMID- 11499285 TI - CMS reaches out to physicians. PMID- 11499286 TI - Convergence and transparency: keys to successful market-neutral performance. PMID- 11499288 TI - Picking up the pieces after a career setback. PMID- 11499289 TI - Why are the Feds hounding this FP? PMID- 11499290 TI - Organized medicine's new turf: e-services. AB - Stodgy medical societies are partnering with, and creating, online companies. What can they do for your practice? PMID- 11499291 TI - Two ways to beat stress. AB - The daily frustrations of medical practice take their toll on body and mind. These techniques can put you back in control. PMID- 11499292 TI - Monitor patients online? AB - New disease management programs are enabling physicians to do just that. Can this help you improve care, or is it just another boondoggle? PMID- 11499293 TI - Financial survey. Retirement funding falls again. AB - Doctors can do more to overcome the lagging pension plan growth reflected in our data. PMID- 11499294 TI - Statutes of limitations may offer a lot less protection. PMID- 11499295 TI - Characteristics and compliance of patients receiving long-term oxygen therapy (LTOT) in Turkey. AB - Compliance is often poor in patients receiving long-term oxygen therapy (LTOT). We assessed some characteristics of LTOT patients and factors associated with non effective use of LTOT in Turkey. A questionnaire was sent to 1100 patients who purchased their oxygen equipment between January 1995 and September 1999 from the largest oxygen company in Turkey. The questionnaire included items about daily duration of oxygen therapy, home situation, lifestyle, education level of the patient, smoking habits and side effects of oxygen therapy. In addition, patients were asked whether supplementary medical and technical education on oxygen therapy was explained to them. 379 patients responded. The patients had been receiving LTOT for 24.3 +/- 13 months. The mean duration of oxygen therapy was 9 +/- 6.8 hours/day. Only 28.2% of the patients managed to receive oxygen therapy for 15 h or more daily. 33.2% of the patients were told to use oxygen only when they had difficulty in breathing. 6.9% of the patients continued to smoke and 8.7% of the patients had complaints related to oxygen therapy. Logistic regression analysis identified that the likelihood of achieving effective oxygen therapy increased 4.5 fold (p < 0.001) when the physician gave pre-treatment education to the patient about the therapy, and effective use increased 3.8 fold (p < 0.002) when the patients were closely followed-up by the physician. We conclude that pre-treatment education of the patient about the therapy and close follow-up by the physician may increase the compliance rate to LTOT. PMID- 11499296 TI - Enhanced lipid peroxidation in cancer tissue homogenates in non-small cell lung cancer. AB - The oxidative modification of nucleic acids by reactive oxygen species may lead to malignant conversion, but its exact role in lung cancer biology is still not clear. Lipid peroxidation, a well-known index of free radicals activity, is a process of oxidative polyunsaturated acids destruction. Our study was aimed to investigate the level of lipid peroxidation ex vivo in tumor tissue and lung parenchyma obtained from patients with non-small cell lung cancer. Thirty-two patients with lung cancer (including 19 with squamous cell lung cancer) were enrolled in the study. During a surgical resection, tumor tissue and lung parenchyma were obtained and the concentration of lipid peroxidation products, i.e. conjugated dienes and lipid hydroperoxides, measured. In the whole group of patients the concentrations of conjugated dienes and lipid hydroperoxides in the tumor tissue were higher than those in lung parenchyma (1.008 +/- 0.503 A233 nm vs. 0.717 +/- 0.283 A233 nm; p < 0.05 and 0.109 +/- 0.062 A532 nm vs. 0.102 +/- 0.087 A532 nm; p < 0.05, respectively). Similar results were obtained in squamous cell carcinoma patients (0.975 +/- 0.348 A233 nm vs. 0.708 +/- 0.300 A233 nm; p > 0.02 and 0.094 +/- 0.029 A532 nm vs. 0.080 +/- 0.071 A532 nm; p < 0.05, respectively). In both groups of patients, a positive correlation between concentration of conjugated dienes in tumor tissue and clinical stage (R = 0.45; R = 0.52; p < 0.05, respectively) was found. Our results confirm the enhanced lipid peroxidation in cancer tissue as compared with matched lung parenchyma. Additionally, a higher level of oxidative stress, expressed as the concentration of conjugated dienes in tumor tissue, was associated with clinical progression of the tumor. PMID- 11499297 TI - Erdheim-Chester disease. A case report. AB - A 63-year old man had a history of diabetes insipidus, arthralgias and myalgias, weight loss, relapsing fever and malaise. Increased uptake of Tc-99m was found predominantly in distal antebrachia, in distal femurs and in both trochanters and tibias on the bone scintigraphy. The chest radiograph showed reticulonodular pattern and the high resolution computed tomography (HRCT) scans revealed diffuse infiltrative lung disease with small multiple nodules and widening of interlobular septs. Videothoracoscopic lung biopsy and biopsy of tibial lesion were perfomed. The histopathologic examination proved non-Langerhans cell histiocytosis-Erdheim-Chester disease. Treatment with prednisone reduced the pain and fever and improved the vital capacity of the lungs while the changes in the lungs and bones remained unchanged. PMID- 11499298 TI - Pulmonary haemorrhage in a patient with Kikuchi disease. AB - Kikuchi disease is an immunologic disease and most commonly presents with cervical lymphadenopathy. It is usually self-limiting [1]. It is sometimes associated with systemic lupus erythematosus (SLE) [2], and can be associated with many complications including aseptic meningitis, neuropathy, brachial neuritis. It can also be associated with lymphoma, and may sometimes be misdiagnosed as such [3]. The following is the first reported case of Kikuchi disease associated with pulmonary haemorrhage. The patient died and no other cause of the pulmonary haemorrhage could be identified at post mortem. PMID- 11499299 TI - Congenital diaphragmatic hernia in an adult simulating a pleural liposarcoma. AB - We report the case of a 32-year-old man with the onset of exercise-related dyspnea, chest pain and chest radiography simulating a pleural effusion. The computed tomography of the chest showed signs suggesting pleural liposarcoma. Because of these findings, a videothoracoscopy was performed which surprisingly showed the presence in the left pleural space of intrapleural omentum and spleen. This report underlines that the clinical manifestations of Bochdalek hernia in adults are variegate and in most cases preoperative diagnosis is not possible. PMID- 11499300 TI - Assessment of the smoker who wants to quit. AB - This paper describes assessments that can be useful when giving smokers help to stop. The assessments discussed are: motivation, amount smoked, dependence, lung function, carbon monoxide (CO), earlier cessation experience, knowledge of nicotine's role, comorbidity, body weight, vital signs and cardiovascular risk factors. The rationale for each assessment is given. The most important factors to assess are probably motivation, dependency and CO in expired air. PMID- 11499301 TI - Photodynamic therapy for treating early stage lung cancer. AB - The importance of treating lung cancer at the earliest stage possible has been increasingly recognised. With the resurgence of interest in lung cancer screening, the use of less invasive staging and treatment procedures has gained momentum. Technical advances have propelled the use of bronchoscopic procedures into clinical practice, providing alternatives for conventional diagnostic and treatment modalities. Many lung cancer patients are limited in their pulmonary reserve capacity due to their smoking history, making them unfit to undergo more invasive medical procedures such as surgery. Surgical resection for lung cancer may also result in a relatively wasteful removal of normal lung parenchyma. In the case of superficial early stage lung cancer in the central airways, a more conserving approach such as surgical bronchoplasty is acceptable. With the move towards early detection of lung cancer, even smaller cancer lesions will be detected. Therefore, these lesions will be suitable for bronchoscopic treatment if nodal disease is absent. The potential of photodynamic therapy and alternative bronchoscopic techniques for the management of early stage lung cancer, as an alternative to, or in combination with, surgical resection should be considered. PMID- 11499302 TI - Bacterial community-acquired pneumonia in children: changes in causative agents. AB - In this review, the authors present the current concept on the bacterial etiology of pediatric community-acquired pneumonia. The changes which have taken place after the introduction of modern serology for the diagnosis of bacterial pneumonia are discussed. Based on the current knowledge, the authors present the recommendations for empirical antibiotic therapy of pediatric community-acquired pneumonia in basically healthy children. PMID- 11499303 TI - Exacerbations of COPD: predictive factors, treatment and outcome. AB - Chronic obstructive pulmonary disease (COPD) is a leading cause of death, and constitutes a major medical and an increasing economic problem for acute and long term care. A low level of irreversible airway obstruction when in stable condition, hypercapnia, hypoxia, the presence of comorbid heart disease, right ventricular failure, and low serum albumin are the main factors related to risk of exacerbations. Bronchial infections, bronchospasm, left ventricular failure, pneumonia, pneumothorax and thromboembolism are described as the most frequent relapsing causes of COPD. During exacerbation, the inflammatory process, the ventilation/perfusion (V'A/Q') mismatching, an increased airflow resistance and dynamic hyperinflation (PEEPidyn) expose the respiratory muscles to the risk of fatigue, eventually leading to ventilatory pump failure and rising hypercapnia. Prevention of exacerbations and subsequent hospitalisations may be obtained with careful rehabilitation programs, a strict drug protocol, long term oxygen therapy and sometimes using home noninvasive mechanical ventilation (NMV). During exacerbation proper management of infection and lung mechanics derangement has to be adopted using an accurate assessment of severity and standardized treatment protocols. Patient history and examination and functional tests are beneficial to decide how and where to treat these patients. Mechanical ventilation (possibly noninvasive) may be required to reverse the acute episode. The aims of all these procedures remain: i) to prolong length and quality of life; ii) to save costs. Both hospital and post-discharge mortality of exacerbated COPD remain high while quality of life appears to be poor. Future studies will elucidate the relation between number and severity of exacerbations and prognosis. PMID- 11499304 TI - Parapneumonic effusion and empyema: best therapeutic approach. PMID- 11499305 TI - Nitric oxide in the regulation of airway tone. PMID- 11499306 TI - Nitric oxide in the regulation of vascular systemic resistance. PMID- 11499307 TI - Nitric oxide and peroxynitrite: the balance between cardioprotection and cardiotoxicity. PMID- 11499308 TI - Nitric oxide and control of pulmonary vascular resistance. PMID- 11499309 TI - Nitric oxide and non steroidal antiinflammatory drugs (NSAID)-related gastroduodenal damage. PMID- 11499310 TI - Nitric oxide in childhood asthma. PMID- 11499311 TI - Exhaled nitric oxide in COPD patients. PMID- 11499312 TI - Nitric oxide in chronic heart failure. PMID- 11499313 TI - Exhaled nitric oxide in patients with alpha 1 antitrypsin (AAT) deficiency. PMID- 11499314 TI - Therapeutic use of nitric oxide in critical settings. PMID- 11499315 TI - Hospital EMTALA liability exposure expanded. Arrington v. Wong. PMID- 11499316 TI - Mississippi Supreme Court allows patient's identity to be revealed. Baptist Memorial Hospital v. Johnson. PMID- 11499317 TI - Physician's alcohol dependence ruled admissible in malpractice action. Watson v. Chapman. PMID- 11499318 TI - A comparative study of three techniques for the analysis of sperm recovery: touch print cytology, wet preparation, and testicular histopathology. AB - PURPOSE: The aim of this study is to evaluate the efficacy of simultaneous testicular touch-print cytology, testicular histopathology, and wet preparation in nonobstructive azoospermic (NOA) males. METHODS: Three hundred and sixty-three males with NOA underwent a multiple testicular sampling prior to ICSI for histopathologic evaluation, diagnostic testicular sperm extraction, and simultaneous touch-print cytology to evaluate sperm presence or absence. A total of 979 testicular samples were taken. RESULTS: Sperm recovery was achieved in 106 cases (29.2%). Patients with hypospermatogenesis and focal spermatogenesis needed 2.8 and 5.9 biopsies, respectively, to retrieve spermatozoa, while in patients with germ cell aplasia and maturation arrest, even after eight to nine samples no spermatozoa were recovered. Neither the FSH levels nor the testicular volume was found to be significant in the prediction of sperm recovery. If only a single testis was to be biopsied, 25% of the cases with sperm recovery would have been missed. The combination of touch-print cytology with histopathology and wet preparation increased the accuracy of spermatozoa identification. CONCLUSION: Touch-print cytology was found to be more predictive than wet preparation in the diagnosis of spermatogenesis; moreover, it was found to be a quick and easy technique providing an accurate diagnosis in prediction of sperm recovery. PMID- 11499319 TI - Evaluation of cryoinjury of spermatozoa after slow (programmed biological freezer) or rapid (liquid nitrogen vapour) freeze-thawing techniques. AB - PURPOSE: This study was initiated to determine the negative effect (cryodamage) on human spermatozoa after freeze-thawing and to find out whether freezing of spermatozoa with a computerized biological freezer is more advantageous than freezing above static liquid nitrogen vapour with regard to spermatozoa vitality, chromatin normality, morphology, and membrane integrity. METHODS: Forty-four semen samples were obtained from patients attending andrology laboratory, and each sample was divided into two aliquots. One aliquot was frozen using static liquid nitrogen vapour (G.II) and the second with a computerized biological freezer (G.III). Acridine orange was used for assessment of chromatin cryoinjury, whereas the morphology was evaluated according to WHO criteria. Hypoosmotic swelling test was used to identify membrane integrity and eosin-nigrosin staining was used to determine the vitality of spermatozoa. RESULTS: The mean percentage of normally condensed chromatin in the native semen sample (G.I) decreased significantly (p < .001) after freeze-thawing by using either liquid nitrogen vapour (G.II), or a biological freezer (G.III), which was significantly higher (p < .001) after freezing with liquid nitrogen vapour than after freezing with the biological programmed freezer. Morphologically normal spermatozoa decreased significantly (p < .001) in both freezing methods in comparison to the native semen samples. In addition, membrane integrity of spermatozoa (HOS-test positive) was significantly lower (p < .001) after the freeze-thawing procedure in G.II and G.III compared to G.I. In both these parameters the deterioration was similar among the two freezing procedures. Finally the mean percentage of live spermatozoa decreased significantly (p < .001) in both freezing techniques in relation to the mean value in the neat semen samples. CONCLUSIONS: Freeze-thawing procedure has a detrimental effect on chromatin, morphology, membrane integrity, and vitality of human spermatozoa not only by freezing above static liquid nitrogen vapour but even by using a computerized biological freezer. However, the chromatin deterioration rates are significantly higher by freezing above static liquid nitrogen vapour in comparison to freezing with a programmed biological freezer. Therefore, we recommend the use of this technique for freezing semen especially when ICSI technique is considered as the main therapeutic procedure. PMID- 11499320 TI - Seasonal changes in human sperm chromatin condensation. AB - PURPOSE: The aim of this study was to investigate possible seasonal changes in human sperm parameters, especially chromatin condensation. METHOD: In a first run, 3155 patients attending the andrological outpatient clinic at the Centre of Dermatology and Andrology at Justus Liebig University, Giessen, Germany, from January 1992 to October 1995 were examined for sperm count, motility, vitality, and chromatin condensation. RESULTS: The respective results were correlated according to season. Significant seasonal changes were observed in chromatin condensation and sperm count, with mean maximum values (for chromatin condensation and sperm count) of 86.24% aniline blue-negative spermatozoa in January and 68.75 x 10(6) mL-1 in April. To confirm the observation of seasonal changes in sperm chromatin condensation in Germany on the Southern Hemisphere, 179 patients attending the Reproductive Biology Unit at Tygerberg Hospital, Tygerberg, South Africa, were examined by means of the aniline blue stain from April 1999 to April 2000. For chromatin condensation, a significant seasonal change shifted by 4-5 months was observed on the Southern Hemisphere. However, no seasonal variations could be found for the sperm count. CONCLUSIONS: Our results clearly demonstrate seasonal changes in sperm count and chromatin condensation. In contrast, no circannual relation was observed for motility and vitality. PMID- 11499321 TI - A prospective, randomized comparison of two commercial media for ICSI and embryo culture. AB - PURPOSE: The aim of this prospective, randomized study was to compare the results obtained in ICSI with two culture media, P-1 (Irvine Scientific) and IVF-50 (Scandinavian IVF Science). METHODS: A total of 182 patients undergoing ICSI treatment were randomly included in this study and divided in two groups: Group I: P-1 medium (n = 91) or Group II: IVF-50 medium (n = 91). All the embryos were transferred on the second day. RESULTS: Patient age did not differ (p = .29) between Group I (34.8 +/- 4.8) and Group II (34.0 +/- 4.5). The number of oocytes retrieved from Group I (10.6 +/- 6.7) was also similar (p = .49) to that retrieved from Group II (11.1 +/- 6.4). In addition, there was no difference (p = .25) in the number of oocytes retrieved at metaphase II between Group I (7.9 +/- 4.6) and Group II (8.7 +/- 4.6). Normal fertilization rates, abnormal fertilization rates, and cleavage rates were similar (p = .62, p = .48, and p = .9, respectively) between Group I (68.4 +/- 23.3%, 6.7 +/- 10.3%, and 98.7 +/- 4.6%) and Group II (65.3 +/- 26.2%, 9.0 +/- 13.8%, and 98.9 +/- 3.9%, respectively). The embryo score was also similar (p = .62) for both groups (Group I: 31.9 +/- 14.0 and Group II: 33.4 +/- 15.8). There was no difference in the number of embryos transferred (p = .69) between Group I (2.8 +/- 1.0) and Group II (2.8 +/- 1.1). In addition, pregnancy rates/puncture, pregnancy rates/transfer, implantation rates, and abortion rates were also similar for Group I (36.2%, 37.0%, 17.4%, and 12.1%, respectively) and Group II (31.8%, 33.7%, 15.8%, and 10.3%, respectively) (p = .64, p = .75, p = .72, and p = 1.0, respectively). CONCLUSIONS: There were no differences in the results obtained with culture media P-1 (Irvine Scientific) and IVF-50 (Scandinavian IVF Science) for ICSI and embryo culture. PMID- 11499322 TI - Increased body mass index has a deleterious effect on in vitro fertilization outcome. AB - PURPOSE: Few studies have addressed the effect of weight on IVF outcome, with some showing a decrease in IVF success and some showing no change in overweight women (BMI > 25 kg/m2) compared to women with normal weight (BMI < 25 kg/m2). METHODS: One hundred thirty-nine women < 40 years old undergoing 180 IVF cycles with fresh embryo transfers were retrospectively evaluated between January 1997 and March 1999, stratified by body mass index (BMI) (cutoff of 25). RESULTS: In the group with BMI > 25 kg/m2, basal FSH, implantation rates (IR), and pregnancy rates (PR) were significantly lower, while the duration of stimulation, gonadotropin requirements, and spontaneous miscarriages were slightly higher, compared to the BMI < or = 25 group. CONCLUSIONS: Excess weight defined as BMI > 25 kg/m2 has a negative impact on IVF outcome. Future prospective studies evaluating oocyte and/or embryo quality, and androgen and insulin levels, between overweight women and those with normal weight are needed. PMID- 11499323 TI - Influence of midluteal serum prolactin on outcome of pregnancy after IVF-ET: a preliminary study. AB - PURPOSE: To investigate the relationship between serum prolactin levels in the midluteal phase and the prognosis of pregnancy after IVF-ET. METHODS: DESIGN: Prospective study; SETTING: Shimane Medical University Hospital; PATIENTS: 29 patients completing their 34th IVF cycle; and MAIN OUTCOME MEASURES: Serum levels of prolactin, estradiol-17 beta, and progesterone in the midluteal phase. RESULTS: Midluteal prolactin levels were significantly lower in patients with early pregnancy loss than in the successful pregnancy group. The mean progesterone/prolactin ratio and the estradiol-17 beta/prolactin ratio were highest in patients with early pregnancy loss and were lowest in patients with successful pregnancies. CONCLUSIONS: Findings of this study suggest that measurement of the serum prolactin concentration in the midluteal phase is useful for predicting the outcome of IVF-ET. PMID- 11499324 TI - Hypothalamic-pituitary suppression with oral contraceptive pills does not improve outcome in poor responder patients undergoing in vitro fertilization-embryo transfer cycles. AB - PURPOSE: To evaluate and compare the use of OCP with GnRHa for hypothalamic pituitary suppression in poor responder IVF patients. METHODS: Retrospective analysis of IVF-ET cycles of poor responders. Hypothalamic-pituitary suppression with OCP (Group I, n = 29) or GnRHa (Group II, n = 52), followed by stimulation with gonadotropin, oocyte retrieval, and embryo transfer. Baseline characteristics and cycle outcomes were compared. RESULTS: 73 women underwent 81 cycles from 1/1/1999 to 1/1/2000. Baseline characteristics were similar. 31/81 (38%) cycles were cancelled (Group I, 14/29 (48%) vs. Group II, 17/52 (33%), NS). Cycle outcomes including amount of gonadotropin, number of eggs retrieved, number of embryos transferred, and embryo quality were similar. Patients in Group I required fewer days of stimulation to reach oocyte retrieval. Pregnancy outcomes were similar in the two groups. CONCLUSION: Our retrospective analysis revealed no improvement in IVF cycle outcomes in poor responders who received OCPs to achieve hypothalamic-pituitary suppression instead of GnRHa. PMID- 11499325 TI - DAZLA: an important candidate gene in male subfertility? AB - PURPOSE: To study the role of the autosomal candidate gene DAZLA (Deleted in AZoospermia Like Autosome) in male subfertility. METHODS: We reviewed clinical data of subfertile men with oligozoospermia or azoospermia, mostly candidates for intracytoplasmic sperm injection (ICSI). Mutation detection was performed using polymerase chain reaction followed by single strand conformation polymorphism analysis. All shifted bands were analyzed by sequencing. RESULTS: We searched for mutations in 44 subfertile men. Nine subfertile men were included, because family history showed that their brothers also faced fertility problems. In these men a possible autosomal gene defect may contribute to their fertility problem. No mutations were found, except for two polymorphisms in intron 4 and 5. CONCLUSION: At this moment it does not seem relevant to search for possible mutations in the DAZLA gene in clinical practice. PMID- 11499326 TI - Growth retardation of rabbit embryos in ligated oviducts and the toxic effects of rabbit tubal fluid on mouse embryo development in vitro. PMID- 11499327 TI - Comparison of treatment of acne vulgaris with alternate-day applications of tazarotene 0.1% gel and once-daily applications of adapalene 0.1% gel: a randomized trial. AB - Tazarotene and adapalene are recently introduced topical retinoids that are useful in the treatment of acne vulgaris. The clinical benefits of each drug have now been compared in a multicenter, double-blind, randomized, parallel-group study involving 164 patients with mild-to-moderate facial acne vulgaris. Patients were randomized to receive 15 weeks' treatment with alternate-day tazarotene 0.1% gel, with vehicle gel on the intervening evenings, or once-daily adapalene 0.1% gel. Both regimens were comparably effective with no significant between-group differences in efficacy measures. A total of 74% of tazarotene-treated subjects and 73% of adapalene-treated subjects achieved at least a 50% improvement in their acne. In addition, there were no clinically significant differences in tolerability. It is concluded that an alternate-day tazarotene regimen offers efficacy and thus tazarotene treatment can be useful even in patients whose compliance may be suboptimal. An alternate-day regimen also offers the potential for considerable savings in drug costs. PMID- 11499328 TI - Randomized facial tolerability studies comparing gel formulations of retinoids used to treat acne vulgaris. AB - Two double-blind, randomized, split-face studies have been performed to compare the facial tolerability of topical retinoids in volunteers with sensitive skin. In one study, subjects applied tazarotene 0.1% gel to one side of their face and tretinoin 0.1% gel microsponge, tretinoin 0.025% gel, or adapalene 0.1% gel to the other side of their face, for up to 29 days. Increases in facial dryness and erythema were comparable among all retinoids. Some subjects in each treatment group experienced levels of retinoid-associated irritation that required temporary suspension of, or reduction in, treatment. Facial dryness and erythema tended to be greater in these subjects than in those who tolerated the regimen without change, suggesting that the need to discontinue or modify treatment depends more on the individual than on any major inherent differences in the irritant potential of these retinoids. A second study compared once-daily versus alternate-day tazarotene 0.1% gel therapy. Tolerability was superior when initiating therapy with the alternate-day regimen. PMID- 11499329 TI - Efficacy and tolerability of once-daily tazarotene 0.1% gel versus once-daily tretinoin 0.025% gel in the treatment of facial acne vulgaris: a randomized trial. AB - Tazarotene 0.1% gel and tretinoin 0.025% gel are both effective in the treatment of acne vulgaris. Results of a multicenter, double-blind, randomized, parallel group study that compared the efficacy and tolerability of these drugs are presented here. A total of 143 patients with mild-to-moderate facial acne vulgaris were randomized to receive tazarotene 0.1% gel or tretinoin 0.025% gel once daily for 12 weeks. Tazarotene 0.1% gel was more effective than tretinoin 0.025% gel in reducing the open comedo count (P < or = .05), the total noninflammatory lesion count (P < or = .05), and the total inflammatory lesion count (not statistically significant). At some time points, tazarotene was associated with increased irritation, but peeling, erythema, dryness, burning, and itching never exceeded trace levels. We conclude that tazarotene 0.1% gel is more effective than tretinoin 0.025% gel in reducing noninflammatory lesions and similarly effective in reducing inflammatory lesions. PMID- 11499330 TI - Safety and efficacy of short-duration oral terbinafine for the treatment of tinea corporis or tinea cruris in subjects with HIV infection or diabetes. AB - Cutaneous fungal infections in immunocompromised patients can be aggressive and difficult to treat. To determine the safety and efficacy of oral terbinafine for the treatment of tinea corporis or tinea cruris in subjects with human immunodeficiency virus (HIV) infection or diabetes, 2 prospective, randomized, open-label studies were conducted in general community and referral centers. HIV positive (n = 6) and diabetic patients (n = 8) between the ages of 18 and 75 years diagnosed with either tinea corporis or tinea cruris, as confirmed by potassium hydroxide (KOH) wet mount microscopy, were randomized to receive either 1 or 2 weeks of the antifungal treatment. Patients received oral terbinafine 250 mg once daily for 1 or 2 weeks. Main outcome measures were mycological cure, determined at week 6 for HIV-positive and diabetic patients. Three subjects were excluded from the efficacy analyses because of negative cultures at screening (n = 2) and lack of follow-up cultures (n = 1). Efficacy results were similar between the 1- and 2-week groups in both studies. All HIV-positive subjects and 83% of diabetic subjects achieved mycological cures at week 6 based on culture results. In a safety population that included all randomized patients (N = 14), no subject experienced adverse events or significant changes in laboratory findings related to study medication. Results of these small series indicate that a short course of oral terbinafine 250 mg once daily is a safe and effective treatment for tinea corporis or tinea cruris in subjects with HIV infection or diabetes. PMID- 11499331 TI - The safety and efficacy of terbinafine in patients with diabetes and patients who are HIV positive. AB - Superficial fungal infections are commonly encountered in dermatologic practices. Their incidence is increasing because of the use of immunosuppressive drugs, an aging population, and the increased prevalence of diabetes mellitus and human immunodeficiency virus (HIV) infection. Topical antifungal therapy typically has been the treatment of choice for uncomplicated dermatophytoses of the skin, such as tinea pedis and tinea cruris. However, these infections may be particularly difficult to treat in high-risk patients, such as those who have diabetes or who are HIV positive. In patients with HIV, dermatophytoses tend to be more extensive and generally require oral antifungal therapy. The allylamine terbinafine has a proven safety record and no significant drug interactions. We review the clinical experience with terbinafine in diabetic and HIV-positive subjects and conclude that terbinafine is safe and has a low drug interaction potential in these high risk cohort studies. PMID- 11499332 TI - Short-duration oral terbinafine for the treatment of tinea pedis in HIV-positive patients. AB - Management of tinea pedis in patients who have the human immunodeficiency virus (HIV) is problematic; in those patients, dermatophytoses may be more difficult to treat than in the general population. This prospective, open-label, multicenter, randomized study evaluated the efficacy and safety of a short course of oral terbinafine for tinea pedis in patients who are HIV positive. Twenty-seven patients were randomized to receive oral terbinafine 250 mg once daily for 2 or 4 weeks; 17 patients with positive initial cultures and follow-up cultures were evaluable for efficacy at week 8. Mycological cure (defined as negative potassium hydroxide [KOH] microscopy and culture results) occurred in 47% (8) of patients; and modified mycological cure (defined as negative follow-up cultures) occurred in 65% (11) of patients. All 27 patients were evaluated for safety. Clinical cure (defined as minimal residual signs and symptoms) occurred in 82% (14) of patients. Oral terbinafine was well tolerated, indicating that regimens of 2 or 4 weeks are safe and effective for the treatment of tinea pedis in patients who are HIV positive. PMID- 11499333 TI - Role of oral antifungal agents for the treatment of superficial fungal infections in immunocompromised patients. AB - Superficial fungal infections or tinea infections (also known as the dermatophytoses) are commonly encountered conditions in clinical practice, affecting the skin, hair, and nails. The most commonly prescribed modality to treat these infections is topical antifungal therapy. However, this method of treating tinea infections may be less convenient and efficacious in the immunocompromised patient. In such patients, skin infections are more difficult to treat because the disease is often more extensive and severe. Tinea infections of the hair and nails usually require oral therapy. Further, topical treatment is not as efficacious as oral antifungal therapy and, with the exception of the topical antifungal agent ciclopirox, is not indicated for the treatment of tinea unguium (onychomycosis). The 2 most frequently prescribed oral antifungal agents to treat onychomycosis are itraconazole and terbinafine. In the general population, both agents are effective in treating fungal nail infections; however, differences in the agents' mechanism of action and metabolic pathways result in differences in efficacy and drug-drug interaction potential. However, limited data exist on the use of these agents in immunocompromised patients for the treatment of onychomycosis and superficial tinea infections. The available efficacy data we have are limited to case reports or small pilot studies; thus, data supporting the efficacy of these agents for the treatment of tinea infections in the immunocompromised patient must be extrapolated from the general population. For safety issues, however, some postmarketing data exist supporting the safety of these agents in the diabetic and human immunodeficiency virus (HIV) patients populations; indeed, both agents appear to be safe. However, one contrasting point between these 2 agents is drug interactions. Oral terbinafine, unlike itraconazole (a potent cytochrome P-450 [CYP] 3A4 inhibitor), has a relatively low potential for drug-drug interactions, making terbinafine a useful agent for the treatment of tinea infections in immunocompromised patients (e.g., those who are HIV positive and those with diabetes), who are likely to be receiving concomitant medications. Further, recently conducted studies of terbinafine for the treatment of tinea pedis, tinea cruris, and tinea corporis infections in these high-risk patient groups also support efficacy claims and reemphasize its relatively safe profile and low potential for drug interactions. Additional studies in other immunocompromised patient populations may be useful to confirm recent studies and expand the potential use for this agent. PMID- 11499334 TI - Births: preliminary data for 2000. AB - OBJECTIVES: This report presents preliminary data for 2000 on births in the United States. U.S. data on births are shown by age, race, and Hispanic origin of mother. Data on marital status, prenatal care, cesarean delivery, and low birthweight are also presented. METHODS: Data in this report are based on more than 96 percent of births for 2000. The records are weighted to independent control counts of births received in State vital statistics offices in 2000. Comparisons are made with 1999 final data. RESULTS: The number of births rose 3 percent between 1999 and 2000. The crude birth rate increased to 14.8 per 1,000 population in 2000, 2 percent higher than the 1999 rate. The fertility rate rose 3 percent to 67.6 per 1,000 women aged 15-44 years between 1999 and 2000. The birth rate for teenagers, which has been falling since 1991, declined 2 percent in 2000 to 48.7 births per 1,000 females aged 15-19 years, another historic low. The rate for teenagers 15-17 years fell 4 percent, and the rate for 18-19 year olds was down 1 percent. Since 1991, rates have fallen 29 percent for teenagers 15-17 years and 16 percent for teenagers 18-19 years. Birth rates for all of the older age groups increased for 1999-2000: 1 percent among women aged 20-24 years, 3 percent for women aged 25-29 years, and 5 percent for women in their thirties. Rates for women aged 40-54 years were also up for 2000. The birth rate for unmarried women increased 2 percent to 45.2 births per 1,000 unmarried women aged 15-44 years in 2000, but was still lower than the peak reached in 1994. The number of births to unmarried women was up 3 percent, the highest number ever reported in the United States. However, the number of births to unmarried teenagers declined. The proportion of women who began prenatal care in the first trimester of pregnancy (83.2 percent) did not improve for 2000, nor did the rate of low birthweight (7.6 percent). The total cesarean rate rose for the fourth consecutive year to 22.9 percent, the result of both a rise in the rate of primary cesarean deliveries and a decline in the rate of vaginal births after previous cesarean delivery. PMID- 11499336 TI - Single-pulse transcranial magnetic stimulation reset the rhythm of essential tremor but not heart beat. AB - BACKGROUND: Human oscillator is observed in and outside the nervous system. Cardiac rhythm is generated by heart itself but can be modulated by brain. Using the technique of transcranial magnetic stimulation (TMS) and resetting index, we studied if single-pulse TMS could reset the cardiac rhythm and help differentiate oscillator of neurogenic or non-neurogenic origin. METHODS: In addition to the study of 4 patients with essential tremor, cardiac rhythm was studied in 6 normal subjects. The magnetic intensity was initiated from motor threshold of hand muscle, and then with an increment of 10% up to the maximal output of magnetic stimulator. We used the resetting index (RI) to quantify the influence of the TMS. RESULTS: The resetting phenomenon was observed in essential tremor (RI = 0.92) but not in cardiac rhythm (RI = 0.02). CONCLUSIONS: Single-pulse TMS is able to reset the rhythm of essential tremor but not heart beat. The pacing mechanism is different between essential tremor and heart beat. The cardiac rhythm is regulated and modulated chiefly by heart itself. Essential tremor should not share the same mechanism with heart beat. PMID- 11499335 TI - Mitochondrial theory of aging matures--roles of mtDNA mutation and oxidative stress in human aging. AB - Mitochondrial theory of aging, a variant of free radical theory of aging, proposes that accumulation of damage to mitochondria and mitochondrial DNA (mtDNA) leads to aging of humans and animals. It has been supported by the observation that mitochondrial function declines and mtDNA mutation increases in tissue cells in an age-dependent manner. Age-related impairment in the respiratory enzymes not only decreases ATP synthesis but also enhances production of reactive oxygen species (ROS) through increased electron leakage in the respiratory chain. Human mtDNA, which is not protected by histones and yet is exposed to high levels of ROS and free radicals in the matrix of mitochondria, is susceptible to oxidative damage and mutation in tissue cells. In the past decade, more than one hundred mtDNA mutations have been found in patients with mitochondrial disease, and some of them also occur in aging human tissues. The incidence and abundance of these mutant mtDNAs are increased with age, particularly in tissues with great demand for energy. On the other hand, recent studies have revealed that the ability of the human cell to cope with oxidative stress is compromised in aging. Comparative analysis of gene expression by microarray technology has shown that a number of genes related to oxidative stress response are altered in aging animals. We discovered that the transcripts of early growth response protein-1, growth arrest and DNA damage-inducible proteins and glutathione S-transferase genes are increased in response to oxidative stress in human skin fibroblasts. Moreover, the activities of Cu,Zn SOD, catalase and glutathione peroxidase decrease with age, whereas Mn-SOD activity increases with age up to 65 years and slightly declines thereafter in skin fibroblasts. Such an imbalance in the function of antioxidant enzymes may result in excess production of damaging ROS in the cell. This notion is supported by the observation that intracellular levels of H2O2 and oxidative damage to DNA and lipids are significantly increased with age of the fibroblast donor. Furthermore, the mitochondrial pool of reduced glutathione declines and DNA damage is enhanced in aging tissues. Taken together, these observations and our previous findings that mtDNA mutations and oxidative damage are increased in aging human tissues suggest that mitochondrial theory of aging is mature. PMID- 11499337 TI - Real-time dynamic telepathology through the Internet: evaluation of a new and economic technology at Taipei Veterans General Hospital. AB - BACKGROUND: Computer-aided, digitalized telepathology was introduced about 10 years ago and is gaining acceptance as a mode of providing pathology to remote site. The usefulness of telepathology for remote diagnosis of histology, cytology and frozen section has been evaluated in USA, Europe and Japan. In Taiwan, the use of telepathology for pathological diagnosis is still rare. In order to evaluate the potential use of this technique, we undertook the study using commercialized and low-cost microscopy, personal computer and software on routine biopsy material. METHODS: Sixty prostatic sextant transrectal needle biopsies, including 30 cases of adenocarcinoma and 30 cases of benign lesions, were retrieved retrospectively. The real-time dynamic images were transmitted to the remote site via internet and reviewed by a senior uropathologist who was unaware of the diagnosis. The diagnoses made at the remote site were correlated to the final diagnoses of the cases. RESULTS: All malignant specimens (30/30) were correctly diagnosed by this method. Only one benign case (1/31) of nonspecific granulomatous prostatitis was misdiagnosed as poorly differentiated carcinoma. CONCLUSIONS: Our results show that this method is a good way for teleconsultation. Further studies on other types of specimen worth encouragement for both intra- and inter-institutional consultation. PMID- 11499338 TI - Correlation between anti-Ro/La titers and clinical findings of patients with systemic lupus erythematosus. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is a complex autoimmune disease with various clinical and serological manifestations. Previous studies have shown the association of SLE and anti-Ro, anti-La and anti-dsDNA autoantibodies with various clinical manifestations. We investigated the relationship between these autoantibodies and clinical findings of our patients with SLE. METHODS: Eighty patients with SLE at Department of Pediatrics, Taipei Veterans General Hospital, Taiwan from October 1999 to March 2000 were enrolled in the study. Frequencies of various clinical manifestations were calculated. Autoantibodies to Ro, La and dsDNA were measured using ELISA method. RESULTS: The most frequent clinical findings were arthritis (92.3%), photosensitivity (90.9%) and malar rash (86.1%). The most frequent laboratory findings were positive anti-nuclear antibody (ANA) (94.9%), low serum CH50 hemolytic titer (88.6%) and positive anti-dsDNA level (72.5%). Patients with anemia or photosensitivity had higher titers of anti-Ro antibody (Ab) than those without the respective clinical findings (p = 0.001 and 0.002, respectively). However, patients with proteinuria had lower anti-Ro Ab titers (p < 0.05). There was no correlation between clinical findings and anti-La Ab titers. Patients have higher titers of anti-dsDNA Ab if they had clinical findings of Raynauld's phenomenon, photosensitivity, arthritis, hypocomplementemia (p < 0.001), thrombocytopenia, proteinuria, or serositis (p < 0.05). No correlation was found between renal involvement and titers of various autoantibodies. CONCLUSIONS: Neither anti-Ro, anti-La nor anti-dsDNA antibodies was correlated with renal involvement in patients with SLE. However, patients with proteinuria had significantly lower titers of anti-Ro Ab than those without proteinuria. PMID- 11499339 TI - In vitro effect of hyperbaric oxygen on the chemolysis of infective stones. AB - BACKGROUND: In order to extend the feasibility of hyperbaric oxygen therapy in the urological field, the present study aimed to investigate the dissolution activity of human infective stones in UROCITRA solution under hyperbaric oxygen condition. METHODS: The dissolution activity of 7 struvite and 11 mixed struvite and carbonate apatite stones in UROCITRA solution were studied under 2.5 atmosphere (atm) hyperbaric oxygen (HBO) status in a Sigma I N-124 monoplace chamber. Another 7 struvite and 10 mixed struvite and carbonate apatite stones were also studied under normal condition. Chemolysis was performed in a drip device with a 150-ml/hour continuous flow rate. RESULTS: Under 2.5 atm HBO status, the PO2 of UROCITRA solution was 365 +/- 44 mmHg, which was significantly higher than that of tap water (113 +/- 62 mmHg) and UROCITRA solution (125 +/- 12 mmHg) under normobaric condition (p < 0.001). The decreases in the stone weight of struvite under normobaric condition were 31 +/- 8.8% after 2 h and 48 +/- 15% after 4 h of treatment. The HBO-enriched UROCITRA solution did not increase the dissolution activity as reflected by comparable decreases in the dried stone weight (31.2 +/- 14.6% and 54 +/- 19% at the 2nd and 4th post-treatment hours, respectively, p > 0.05). Similarly, there was no significant difference in the percent stone weight decrease of the mixed struvite and carbonate apatite stones under either HBO or normobaric condition. The dissolution responsiveness of struvite was significantly greater than that of the mixed struvite and carbonate apatite stones. CONCLUSIONS: The chemolysis of struvite in UROCITRA solution is significantly greater than that of the mixed struvite and carbonate apatite stones. However, the UROCITRA solution enriched with HBO does not enhance the dissolution of infective stones. PMID- 11499340 TI - Esophageal cancer after endoscopic injection sclerotherapy for esophageal varices. AB - We reported two cases of squamous cell carcinoma of the esophagus following endoscopic injection sclerotherapy (EIS) for esophageal varices. Both patients were cigarette smokers and had a long history of alcohol abuse. HBsAg and Anti HCV were negative, and Anti-HBs was positive in one of the patients. They were diagnosed as alcoholic cirrhosis with esophageal varices and received EIS treatment. Sotradecol was utilized as the sclerosant with a mean total volume of around 30 ml. Patients developed dysphagia at 5 and 48 months following EIS, respectively. Endoscopic examination showed stenosis and ulcerative mass at the lower portion of the esophagus. Biopsy revealed well- to moderately differentiated squamous cell carcinoma of the esophagus. We conclude that endoscopic follow-up is essential and carcinoma of the esophagus should be included in the differential diagnosis for esophageal ulceration and dysphagia following EIS, particularly in those patients with risk factors for developing esophageal carcinoma. PMID- 11499341 TI - Solitary fibrous tumor of the liver. AB - Solitary fibrous tumor of the liver is a rare neoplasm. So far, 21 cases have been described in the English literature. We reported an additional case. Our patient, a 75-year-old man, suffered from abdominal fullness and body weight loss of 6 kg over 6 months, and chills and fever for 2 months. Hypoglycemia was noted at admission. Both abdominal sonography and CT showed a huge mass occupying the right lobe of liver. Liver biopsy showed fibrous tumor. Right lobectomy was performed and the tumor was resected. Pathological examination showed spindle shaped and fibroblast-like cells within the collagenous stroma. On immunohistochemical stains, these spindle tumor cells showed CD34 positive reactivity. The post-operative course was uneventful and there was no more hypoglycemia. The patient recovered smoothly, regained his body weight, and was alive without evidence of disease recurrence at the last follow-up visit in November, 2000. PMID- 11499342 TI - Hyperbaric oxygen for carbon monoxide poisoning-induced delayed neuropsychiatric sequelae. AB - The clinical manifestations of delayed neuropsychiatric sequelae after carbon monoxide (CO) intoxication are variable. In addition, there is no specific therapy for these complications. Fortunately, these complications have occurred less frequently in recent years, probably due to the usage of hyperbaric oxygen (HBO) therapy. We report an 8-year-old boy who developed late psychiatric disturbances 2 days after full recovery of consciousness from initial CO intoxication. His neuropsychiatric symptoms included consciousness disturbance, motor dysfunction, chorea, aphasia and agnosias. He received HBO therapy at 2.0 barr for 60 minutes once a day for 7 consecutive days. Three weeks later, he was functioning normally with no neuropsychiatric symptoms. A literature review concluded that HBO may be effective in treating neuropsychiatric sequelae. Moreover, immediate administration of HBO during acute CO intoxication may prevent these complications. PMID- 11499343 TI - Buried bumper syndrome--complication of percutaneous endoscopic gastrostomy. AB - Percutaneous endoscopic gastrostomy has gained wide acceptance as a relatively safe and efficient means of long-term enteral nutrition support. We describe an elderly patient in whom the internal bumper eroded into the gastric wall and was completely covered by gastric mucosa about 2 months after gastrostomy tube placement. The end orifice of the gastrostomy tube in the stomach lumen was patent, so it functioned well. Needle-type papillotome was applied endoscopically to cut the overlying mucosa, explored the buried bumper and then, reverted the gastrostomy tube in situ. We reviewed the reported methods and recommended this approach for patients with "buried bumper" syndrome to prevent continued tube migration into the gastric wall. PMID- 11499344 TI - [Twenty years of in vitro fertilization: realization and questions for the future]. AB - IVF is a well-established procedure for the treatment of longstanding infertility due to tubal disease, endometriosis, unexplained infertility or certain types of infertility involving a male factor. IVF can also be applied to couples requiring oocyte donation. Conventional IVF involves several related procedures: patient selection, ovarian stimulation, oocyte retrieval, semen preparation, insemination of cumulus-oocyte complexes, assessment of fertilization, assessment of embryo cleavage, replacement to the uterus of mostly two or three embryos, cryopreservation of excess embryos and establishment of pregnancy. GIFT and ZIFT are related techniques, suitable in couples with at least one healthy Fallopian tube. Conventional IVF is a potentially successful procedure in patients with tubal and unexplained infertility. However, fertilization may fail in couples with certain forms of andrological infertility, especially those in which sperm function is severely deficient. Techniques of assisted fertilization--partial zona dissection (PZD) and subzonal insemination (SUZI)--have been used with limited success to treat couples with severe andrological infertility who could not be helped by conventional IVF. ICSI--the injection of a single spermatozoon into the cytoplasm of a fertilizable metaphase II oocyte--has proved to be more efficient than PZD and SUZI for the alleviation of severe male-factor infertility. Nowadays ICSI can be considered as an infertility treatment, providing similar results in male infertility as conventional IVF in female factor or idiopathic infertility. The ICSI treatment involves several related steps: selection of patients for ICSI, ovarian stimulation and oocyte handling, evaluation and preparation of spermatozoa, the ICSI procedure itself, oocyte damage and pronucleus formation after ICSI, embryo development and replacement and cryopreservation of excess embryos. The audit of IVF and ICSI results is hampered by the way these data are registered in different countries and different centers. A correct and reliable analysis is possible in the United Kingdom; the UK data bank was used to study the factors influencing the outcome of IVF--especially the female age has a determining role. Infertility treatments have induced a sharp increase in the number of multiple pregnancies. Reducing the number of multiple gestations should be considered as a future major challenge for all infertility centers. Since its introduction in 1991 questions have been raised concerning the safety of ICSI, a novel and efficient assisted fertilization procedure. A careful follow-up of the ICSI pregnancies and children is therefore indicated. At the VUB this prospective follow-up of genetic counseling, the possibility of prenatal diagnosis, the problems during pregnancy, the occurrence of (major) congenital malformations as well as a further medical and psychomotor follow-up. The results of 1437 fetal karyotypes indicate that in comparison with a control group there is after ICSI a slight but significant increase of sex chromosomal aneuploidies and de-novo structural aberrations. The percentage of ICSI children (n = 2840) with major congenital malformations was similar to the malformation rate in children from conventional IVF (n = 2955) or natural conception. This prospective follow-up study should be continued much longer--if possible until adulthood to assess the fertility of these children. PMID- 11499345 TI - [From pill to pill: oral contraception, venous thrombosis and coagulation disorders]. AB - "The pill" (oral contraception) is one of the most frequently used and efficient means of birth control. The pill was developed in the middle of the 1950s, and was prescribed in Europe from the early sixties onwards. Because millions of healthy young women in industrialized societies use oral contraceptives, even rare side effects receive a lot of attention. In the early 1960s "the pill" was associated with an increased frequency of arterial and venous thrombosis. However rare, these findings often engendered strong controversies. One of the reasons thereof was the uncertainty about the mechanism: it was not understood how the pill could cause an increased tendency towards thrombosis. In the past 5 years, epidemiologic, haemostatic and clinical research has led to new insights in how "the pill" causes venous thrombosis. These developments followed the discovery of "Activated protein C resistance" and the factor V Leiden mutation. The developments were accelerated by the finding that "third generation oral contraceptives" are associated with a higher risk of venous thrombosis. The lecture describes these new scientific insights. PMID- 11499346 TI - Prognostic factors in stage I ovarian carcinoma. AB - Most studies on prognostic factors in stage I ovarian carcinoma have been hampered by a relative small number of patients included. In this study we identified the most important independent clinical and pathological prognostic factors in stage I epithelial invasive ovarian carcinoma in a large data base of 1545 patients with stage I epithelial ovarian carcinoma. The patients were treated in 6 different countries but were analysed in the same way. Because of the increasing use of endoscopy to remove possibly malignant cysts and the reports on rapid spread of ovarian carcinoma in the peritoneal cavity after laparoscopic removal of ovarian cancers, special attention was made to the presence and timing of the cysts before or during surgery. The multivariate analyses identified degree of differentiation as the most powerful prognostic indicator of disease-free survival, followed by rupture before surgery, rupture during surgery, International Federation of Gynaecology and Obstetrics (FIGO) 1973 stage and age. When the effects of these factors were accounted for, none of the following were of prognostic value for disease-free survival: histological type, dense adhesions, extracapsular growth, ascites, FIGO stage 1988, and size of tumour. In conclusion, degree of differentiation was the most powerful prognostic indicator in Stage I ovarian cancer and should be used when deciding therapy in clinical practice. We also strongly advocate the inclusion of degree of differentiation in a new FIGO classification of stage I ovarian carcinoma. In addition, rupture before and during surgery, FIGO Stage 1973 (Ib versus Ia) and age were shown to be independent prognostic factors. Hence, every effort should be made to avoid rupture during primary surgery of malignant ovarian tumours confined to the ovaries. PMID- 11499347 TI - Continuous quality improvement in public health in Ghana: CQI as a model for primary health care management and delivery. AB - This article describes the context and examines factors influencing the quality of primary health care delivery and management in Ghana, West Africa. It describes the potential of continuous quality improvement as a management philosophy and tool to improve the quality of primary health care delivery and management in Ghana. It compares the Ghanaian context in which the use of continuous quality improvement in health care is new and untried with the developed country context (mainly the U.S.), where continuous quality improvement has a relatively long history of use in health care and is a requirement for the accreditation of health institutions. Finally, the article discusses the steps that have to be taken to translate continuous quality improvement from a theoretical management concept to improve quality of care to an actual managerial intervention in Ghana. In conclusion, continuous quality improvement is shown to be a potential viable approach to improving quality of care in the Ghanaian context and merits further investigation. PMID- 11499348 TI - Community-based health improvement: lessons from the Learning for Action Institute, Simmons College. AB - The Learning for Action Institute at the Graduate School for Health Studies, Simmons College, uses continuous quality improvement concepts and methods to help community-based groups to make positive change. Eight core concepts are the basis for creating sustainable improvements, developing new models for community capacity, and building and disseminating knowledge of what has been learned. These concepts are: clarify the aim, form the right team, target improvement efforts, use data, listen to the customer, use tools and methods, conduct improvement and learning cycles, and make improvements. Two community-based projects are used to illustrate the concepts. PMID- 11499349 TI - School-based health centers as a locus for community health improvement. AB - School-based health centers are critical resources for providing and coordinating health and medical services for children and adolescents. As such, they are an increasingly important component in a strategy to meet the comprehensive health, social, and educational needs of students and families. We show how educators and health professionals, using the language, methods and principles of continuous improvement, can collaborate effectively in addressing the specific concerns of school attendance and teen smoking. PMID- 11499350 TI - Proving the effectiveness of community programs in jeopardy: the role of formative evaluation. AB - Today's health care programs are being scrutinized like never before and yet few clinicians and managers know how to show the value of programs without time consuming outcomes research. This is especially true for new and innovative programs that are dependent on outside funding and may not have enough time to show an effect. However, even well established programs usually lack a clear operational model (clear aim, well defined target population, objectives, performance measures) and well-defined theory (plausible link between activities and desired outcomes). This article proposes a methodology to assure that programs have all the components needed to assure success. PMID- 11499351 TI - The use of volume standards in health services. AB - The ways in which volume standards are implemented by health services organizations are not clear. Therefore, the authors sought to evaluate the extent of use of volume standards, the purposes for which such standards were developed, and the sources of the standards in a sample of health services organizations. The authors found that volume standards were used widely by accrediting organizations, professional societies, and hospitals in their sample, but almost never by health maintenance organizations. Volume standards were used for ensuring adequate experience among residents, providing guidelines to residency programs, and privileging and credentialing physicians. Expert consensus appeared to be the usual source of volume standards. PMID- 11499352 TI - Environmental and organizational determinants of quality management. AB - The aim of the study was to obtain more insight into the organizational and environmental determinants of the implementation of quality management in health care organizations. Primary survey data were collected in 1995 in a large nationwide study within 15 fields of health care and health care-related social services in The Netherlands. In general, there are more differences between health care organizations than between fields of health care. Environmental influence was found to be less important than was expected. Care-oriented organizations have a greater opportunity to involve their patients. It seems that apart from patients, the perceived pressure from other third parties has little influence on the implementation of quality management in health care organizations. PMID- 11499353 TI - QMHC Interview. Interview with Dean Rosen, JD. PMID- 11499354 TI - Continuous alloantigen elimination--a concept for allograft tolerancer. AB - The "enhancement" phenomenon evoked in rats by administration of donor lymphocytes and recipient-anti-donor-lymphocyte antibodies prior to organ transplantation brings about "continuous alloantigen elimination". This new concept is based on the experimental data from studies on the distribution of radiolabelled alloantigen and alloantibody in the recipient lymphoid organs and suggests that immediately after organ transplantation to the "enhanced" recipient, the circulating antibodies bind to the donor transplantation antigens both on endothelial cells and those shed into recipient blood circulation. The formed alloantigen-alloantibody complexes are trapped in the spleen and after opsonization and binding to the migrating splenocytes are transported to the liver for further processing. The process of continuous binding of alloantigen and elimination as alloantibody-alloantigen complexes in the lymphoid organs of the recipient prevents activation of the recipient effector cells and their migration to the graft. The organ grafts survive above 100 days. Although there is no evident rejection in the allograft itself a phenomenon of "rejection alert" is seen in the lymphoid tissue. The lymphocytes originating from the organ graft donor injected intravenously are rejected in the "enhanced" recipient spleen and nodes within 6 hours. The "physiological" site of elimination (rejection) of alloantigens are the lymphoid organs, as it happens with invading pathogens. The allotransplant is spared since its alloantigens are "blocked" by anti-donor specific antibodies, the complexes are continuously detached and washed-off by the percolating blood and eliminated in the lymphoid tissues and liver. PMID- 11499356 TI - Significant improvements in the management of immunosuppression over the last decades have been a major impetus for the expansion of organ transplantation. PMID- 11499355 TI - The effect of short G-CSF administration on the numbers and clonogenic efficiency of hematopoietic progenitor cells in bone marrow and peripheral blood of normal donors. AB - We have analysed the cellularity, the number of clonogenic cells and their clonogenic efficiency (the number of clonogenic cells/2 x 10(5) MNC) in peripheral blood (PB) and bone marrow (BM) during and after filgrastim (rhG-CSF) mobilization of CD34+ cells in 12 healthy donors for allogeneic stem cell donation. G-CSF was administrated subcutaneously for 5 consecutive days at a dose of 10 micrograms/kg/day. WBC, MNC, CD34+ cell counts, CFU-GM and BFU-E assays in PB were performed at baseline and then daily 12 hours after each G-CSF dose. BM was assayed before start (day 1) and after the last dose (day 6) of G-CSF. Results are given as medians, with ranges in parentheses. In PB the total WBC and MNC increased 7.4-fold (6.0-12.3) and 3.3-fold (1.5-9.4), respectively, reaching a peak of 49.4 x 10(9)/l (32.5-66.6) on day 6 for WBC and 6.28 x 10(9)/l (4.7 13.3) for MNC on day 5. CD34+ cell number reached a peak value of 48.0 x 10(6)/l (45.6-285) on day 6 whereas CFU-GM and BFU-E reached their peaks on day 5, 0.95 x 10(4)/ml (0.05-6.08) and 1.04 x 10(4)/ml, respectively. CFU-MIX, not detectable at baseline, reached a peak of 0.95 x 10(4)/ml (0.006-0.51) on day 5 as well. This was accompanied by an increase in CFU-GM, BFU-E and CFU-MIX clonogenic efficiency: 23-fold (3-150), 9.75-fold (2.2-27.8) and 20-fold (2.5-210), respectively. In BM the total WBC number increased 2.5-fold (1.3-4.9) from the baseline value of 52.6 x 10(9)/l (7.9-137.0) whereas the MNC count increased 2.0 fold (0.81-3.7) from a baseline of 13.6 x 10(9)/l (3.5-54.8). This was, however, not significant. The number of CD34+ cells increased significantly 2.9-fold (0.8 8.3). In 8 donors CFU-MIX were detectable before but not after G-CSF treatment. A similar decrease in CFU-GM and BFU-E clonogenic efficiency occurred but was not significant. CFU-GM and BFU-E numbers did not change. We conclude that the total body numbers of lineage committed progenitors increased during G-CSF administration, which indicate their proliferation in addition to mobilization. The effect of G-CSF on the number of more primitive progenitors in BM is less clear and needs further investigation. PMID- 11499357 TI - Molecular and cellular events associated with ischemia/reperfusion injury. AB - I/R is an important non-specific, antigen independent event, which significantly influences the outcome of transplanted organs. Increasing graft immunogenicity and host alloresponsiveness inflicts additional deleterious effects. Ischemia has also been associated with donor conditions such as brain death and the non-heart beating donor. As an event surrounding organ procurement, preservation and revascularization occurring early in the transplant process, it initiates a cascade of molecular and cellular events which trigger the release of proinflammatory mediators and attraction of various cell types infiltrating the tissues. This inflammatory event influences both acute functional and structural changes in the organ which contributes to reduced graft survival. Eventually, attenuation of I/R by strategies targeting various mediators and cell populations at different levels of the inflammatory cascade may constitute a means to improve both short and long-term success of solid organ grafts. PMID- 11499358 TI - Brain death and myocardial energy metabolism: in vivo and ex vivo phosphorus magnetic resonance spectroscopy studies on the feline donor heart. PMID- 11499359 TI - Mitochondrial energy metabolism in brain-dead organ donors. AB - OBJECTIVES: Change from aerobic to anaerobic metabolism has been described in brain-dead organ donors (BDOD) managed for organ procurement. Such modifications could lead to a depletion in intracellular adenine nucleotides and in part explain primary graft dysfunction. The purpose of this study was to investigate the mitochondrial energy metabolism in BDOD using permeabilized muscle fibers. METHODS: With institutional approval, the study was performed prospectively in 15 consecutive BDOD. In each patient, muscle biopsy and blood samples for the determination of plasma lactate and pyruvate were obtained just before organ removal. Mitochondrial respiratory parameters of skeletal muscle were determined in saponin-skinned muscle fibers. Mitochondrial oxygen consumption rates were measured polarographically using different substrates and inhibitors of the respiratory chain complexes. Results were compared to those obtained from muscle biopsies performed in 10 healthy patients during orthopedic surgery. RESULTS: Fifteen donors, 13 men and 2 women, aged 35 +/- 11 yrs, were studied. All patients had a high lactate-to-pyruvate ratio (10). Mitochondrial respiration rates were significantly decreased during adenosine triphosphate (ATP) synthesis. CONCLUSIONS: Major changes in energy metabolism occurred during brain death with a decrease in ATP synthesis capacity. High-risk donors should be recognized for a better graft evaluation. PMID- 11499360 TI - Myocardial performance after brain death: studies in isolated hearts. AB - OBJECTIVES AND METHODS: Brain death related hemodynamic instability and/or cardiac dysfunction is frequently described in the potential organ donor which may lead to exclusion of the heart from transplantation. The underlying mechanisms are controversely discussed. Therefore, in the present study, potential brain death associated cardiodepressant factors were evaluated separately in cross-circulated canine heart models. Brain death was induced by inflation of a subdural balloon catheter. Loading conditions and coronary perfusion pressure were kept identical in all cross-circulated hearts throughout the experiment. RESULTS: Induction of brain death led to a significant hyperdynamic response in all groups, with a maximal effect by the combination of neural and humoral pathways. After the initial reaction all hemodynamic parameters returned to baseline and remained stable until the end of experiments. Even if the hearts were explanted from brain dead donors with typical hemodynamic deterioration in vivo, they showed no significant differences in comparison to the other groups including healthy controls ex vivo. CONCLUSIONS: Therefore we conclude, that hemodynamic instability in the potential donor may rather reflect altered loading conditions and impaired coronary perfusion than neuro-humorally mediated direct myocardial injury. PMID- 11499361 TI - Leukocytes and endothelium interaction as rate limiting step in the inflammatory response and a key factor in the ischemia-reperfusion injury. AB - Leukocyte-endothelium interactions play a key role in regulation of the inflammatory response, leukocytes migration and ischaemia-reperfusion injury. These adhesive reactions controlling the circulation of leukocytes, are key parts of immune surveillance arising from extravasation of neutrophils, and migration into tissue to eliminate invading microorganism. They also play important role in the generation of ischaemic-reperfusion injury of different organs including brain. Plasma levels of soluble adhesion molecules may be a diagnostic marker of the systemic endothelial injury. It is likely that the next few years bring new therapies to control leukocyte-endothelial interaction by directly inhibiting the adhesion molecules or by modulating their expression. PMID- 11499362 TI - Evaluation of donor cardiac function for heart transplantation: experience of a French academic hospital. AB - OBJECTIVES: The diagnostic and therapeutic approaches for evaluation and management of cardiac function in brain-dead patients vary from country to country. The aim of the present study was to describe the results of the evaluation of brain-dead patients as potential cardiac donors in a French teaching hospital that manages the largest number of brain-dead patients in France. METHODS: Demographic parameters, the causes of brain death, clinical evolution, hemodynamic parameters, doses of inotropic and/or vasopressive drugs, the results of echocardiographic examination, and several biochemical markers of myocardial cell injury were retrospectively collected. RESULTS: Seventy-one consecutive brain-dead patients admitted to the intensive care unit of the Academic Hospital of Nancy from October 1st, 1998 to September 30, 1999 were analyzed. Twenty-nine patients were considered as potential heart donors: 22 males and 7 females aged 33 +/- 3 years (Mean + SEM). The cause of brain death was head trauma in 17 cases (59%), cerebrovascular disease in 10 cases (34%), and cerebral anoxia related to cardiac arrest in 2 cases (7%). Eighteen hearts (18/29 or 66%) were harvested and transplanted with a favorable outcome at one month in 17 cases. In 11 cases, the heart was not harvested, nine (9/29 or 31%) because of myocardial dysfunction upon subsequent echocardiographic examination and 2 because of the lack of matched recipients. CONCLUSION: Comparison of these results with those of other groups suggests that hormonal substitution with insulin and triiodothyronine in the presence of myocardial dysfunction could be of potential interest to correct myocardial dysfunction and increase the number of donor hearts. PMID- 11499363 TI - Consequences of labetalol administration on myocardial beta adrenergic receptors in the brain dead pig. AB - OBJECTIVES: Cardiac dysfunction following brain death is associated with highly increased myocardial norepinephrine, lactate and adenosine concentrations. Administration of labetalol, a mixed alpha-, beta-adrenergic receptor antagonist, attenuates metabolic disturbances and improves myocardial function. The purpose of this study was to investigate beta-adrenergic receptor (beta AR) density and affinity in the presence or absence of labetalol administration, as a possible mechanism of the protective effects of this drug. METHODS: Experimental animals were divided into three groups: sham-operated, brain-dead pigs, and brain-dead pigs treated with labetalol (10 +/- 3 mg/kg). The maximum number of binding sites (Bmax) and the dissociation constant (Kd) of beta AR were determined with (-) [125I]cyanopindolol on myocardial samples harvested 3 hours after brain death. RESULTS: Left ventricular beta AR density and affinity were identical in brain dead and sham-operated animals. Labetalol-treated pigs exhibited a significant decrease of Bmax and an increase of Kd as compared with brain-dead pigs. Bmax decrease was due to the persistence of labetalol in the membrane preparations. Increased Kd was too low to be biologically significant. Therefore, beta AR number and affinity can be considered as unchanged after adrenergic blockade with labetalol. CONCLUSIONS: The protective mechanism of labetalol on brain death induced myocardial dysfunction cannot be explained by changes in beta AR density and affinity but is probably related to a preservation of the oxygen consumption/oxygen delivery balance during the autonomic storm. PMID- 11499364 TI - The influence of donor brain death on short and long-term outcome of solid organ allografts. AB - Long-term survival rates of solid organ allografts have improved relatively little during the transplant experience despite more effective immunosuppression, better organ preservation techniques and advances in perioperative management. Because grafts of potentially diminished quality are increasingly accepted to reduce the severe shortage of organs, it has become apparent that a variety of donor-associated risk factors may influence adversely their short and long-term outcome. Recent interest has focused particularly on systemic changes occurring after donor brain death (BD). Numbers of experimental and clinical studies have elucidated the complexities of the hemodynamic, metabolic, neurohormonal, and other physiological alterations following this devastating central injury. This article will address the potential derangements in peripheral organs which may influence their behavior after transplantation. PMID- 11499365 TI - Medical education. The tomorrow people. PMID- 11499366 TI - Medical education. School daze. AB - Most US medical schools have adopted programmes of affirmative action but these have not been completely successful in increasing diversity. There have been some law suits against medical schools brought by rejected students claiming reverse discrimination. Most minority groups are experiencing a downturn in representation in medicine. The movement to humanize medicine through widening entrance criteria has faded away. PMID- 11499367 TI - Medical education. Early learning. PMID- 11499368 TI - Freaky fungi in Japan. Authorities are tripped up by 'magic mushrooms'. PMID- 11499369 TI - The career of a celebrity pill. As Prozac's long reign comes to an end, experts are questioning its legacy. PMID- 11499370 TI - A curse and a cure. Is the crackdown on an abused drug causing needless suffering? PMID- 11499371 TI - Virtual colon scans eliminate much of the discomfort of this important test. PMID- 11499372 TI - Stevia adds sweetness, but don't call it a sweetener. PMID- 11499373 TI - Trial patients need to ask questions after Hopkins death. PMID- 11499374 TI - Estranged relatives can repair broken bonds. PMID- 11499375 TI - Travel woes can be eased when insurance picks up some of the cost. PMID- 11499376 TI - [NVT/VMTI Congress: Risks in the dental profession]. PMID- 11499377 TI - Take two aspirin and hit the send key. Doctor e-mail. PMID- 11499378 TI - Chest X-rays. PMID- 11499381 TI - [Gastroduodenocolonic fistula developing for 45 years after cholecystectomy]. PMID- 11499382 TI - [Choledochal cysts]. PMID- 11499383 TI - Trends and milestones: Large state residential facilities: status and trends in population characteristics as of June 30, 2000. PMID- 11499384 TI - Preface. Long-term outcome following severe brain injury. PMID- 11499385 TI - Overexpression of MnSOD protects murine fibrosarcoma cells (FSa-II) from apoptosis and promotes a differentiation program upon treatment with 5 azacytidine: involvement of MAPK and NFkappaB pathways. AB - Stable transfection of neomycin and human manganese superoxide dismutase (MnSOD2) expression plasmids into a murine fibrosarcoma cell line (FSa-II) was previously done in our laboratory. Treatment with 10 microM 5-azacytidine induced apoptosis in the control cell line (NEO), whereas the MnSOD-overexpressing cell line (SOD H) demonstrated differentiated-appearing morphology. The levels of the myogenic transcription factor, MyoD, and the muscle-specific marker, alpha-actin, were increased over time with 5-azacytidine treatment in the SOD-H cell line. Nuclear transcription factor NFkappaB was activated in the SOD-H cell line, whereas inhibition of NFkappaB activation reduced the levels of MyoD and alpha-actin. Members of mitogen-activated protein kinase pathway and the Raf1/MEK/ERK cascade were shown to play a positive role in this event. Overexpression of MnSOD not only can protect cells from the toxic effects of 5-azacytidine, but can also promote the fibrosarcoma cells to enter a differentiation program. PMID- 11499386 TI - [Pharmacological study of Ajuga iva]. PMID- 11499388 TI - Conference report. Direct adhesive materials: current perceptions and evidence- future solutions. AB - This paper reports the proceedings, including the consensus views of an Australasian expert group convened to consider current perceptions, evidence and future solutions in the field of direct adhesive materials. This group, in anticipating a trend to preservative dentistry, formed the view that caries risk assessment should increasingly influence the selection of restorative materials. In low caries risk patients, aesthetic demands typically favour the use of resin based composites. Interactive (biomimetic) materials based on glass-ionomer chemistry have particular application in high caries risk patients. Teaching in dental schools, continuing education programmes and research, both in the laboratory and in the clinical environment should be more attuned to the ever increasing use of tooth-coloured restorative materials in everyday clinical practice. Linked to this trend are changes in patient--dentist relationships, whereby patients should be encouraged to become more involved in treatment decision-making. Expert group meetings are suggested to be of value in addressing some of the shortfall between the need for good evidence and the relentless challenge of the introduction of new products and concepts in the field of direct adhesive materials. PMID- 11499389 TI - [Onychomycoses: the advantage of a certain diagnosis]. PMID- 11499390 TI - Somatostatin plus isosorbide mononitrate versus somatostatin in the control of acute gastro-oesophageal variceal bleeding: a double blind, randomised, placebo controlled trial. PMID- 11499391 TI - Dilation of benign strictures following low anterior resection using Savary Gilliard bougies. Endoscopic treatment of benign anastomotic colorectal stenosis with electrocautery. PMID- 11499392 TI - Separate Nocardia infections in a patient with chronic granulomatous disease. PMID- 11499393 TI - Meningitis due to Streptococcus salivarius. PMID- 11499394 TI - Helicobacter pylori in children: acquisition of antimicrobial resistance after an initial course of treatment. PMID- 11499395 TI - LightCycler-based quantitative PCR for rapid detection of human herpesvirus 6 DNA in clinical material. PMID- 11499396 TI - Diagnosis of catheter-related bacteremia in cancer patients. PMID- 11499397 TI - Description of Mycobacterium heckeshornense sp. nov. PMID- 11499398 TI - Under one roof: the Miami Project to Cure Paralysis model for spinal cord injury research. AB - Concentrating a wide range of spinal cord injury (SCI) research laboratories in a single location to accelerate progress and draw attention to the promise of SCI research has made The Miami Project to Cure Paralysis one of the most publicly recognized and often controversial research groups in the neurosciences. A "Center of Excellence" at the University of Miami School of Medicine, the Miami Project also serves as a model for SCI research programs being developed nationally and internationally. Founded in 1985, the Miami Project set out on an unprecedented path-to develop a research center dedicated to improving treatments for SCI by bridging basic and clinical science. In doing so, neurosurgeon Barth Green, M.D., enlisted not only a multidisciplinary team of scientists but also a devoted following of financial donors and volunteer research subjects, and support from the University of Miami and Florida legislature. Highly visible spokespersons, including cofounder ex-Miami Dolphin Nick Buoniconti and his son Marc, brought the issue of SCI paralysis and the promise of research before the public, the media, and sports communities. As progress in the neurosciences has raced ahead, public attention to medical research, and SCI research in particular, has grown exponentially. This review will assess the Miami Project as a model for disease-based research that unites academic, philanthropic, and patient communities in a common cause. PMID- 11499399 TI - Neuroscience curricula for undergraduates: a survey. AB - Directors of graduate and undergraduate programs in neuroscience were asked to describe the optimal preparatory background of undergraduates entering graduate programs in neuroscience. Both undergraduate and graduate directors ranked research experience as the most important credential. Graduate directors considered grades/grade point averages and graduate record examination scores as the second and third most important variables. Undergraduate directors ranked courses and grades/grade point averages as their second and third choices. The most essential course for both types of directors was introduction to biology. The next most essential courses for the undergraduate directors were organic/inorganic chemistry and introduction to neuroscience (tied percentages), whereas the graduate directors chose calculus. This latter choice was interpreted as a symbolic representation of the need for quantification in science, generally. Both types of directors chose a course in biochemistry as the most important or desirable course but not necessarily an essential course. These findings have important implications for the training of future undergraduate neuroscience majors. PMID- 11499400 TI - Serotonergic neuromodulation in the cerebellar cortex: cellular, synaptic, and molecular basis. AB - The cerebellum, like most sensorimotor areas of the brain, receives a serotonergic innervation from neurons of the reticular formation. It is well established that local application of serotonin modulates the firing rate of cerebellar Purkinje cells in vivo and in vitro, but the mechanisms by which serotonin affects the cerebellar function are still poorly understood. Whereas interactions between serotonin, glutamate, and GABA have been reported to increase or decrease the firing frequency of Purkinje cells, there is little evidence for a modulation of excitatory and inhibitory synapses by serotonin in the cerebellar cortex. Changes in the intrinsic electrical properties of Purkinje cells upon application of serotonin have also been reported, but their impact on Purkinje cell firing is unclear. The recent finding that serotonin specifically modulates the activity of Lugaro cells, a class of inhibitory interneurons of the cerebellar cortex, offers new insights on the action of this neuromodulator. The peculiar axonal projection and specific interneuronal targets of the Lugaro cells suggest that the action of serotonin might occur upstream of Purkinje cells through a resetting of the computational properties of the cerebellar cortex. Understanding the mechanisms of the serotonergic modulation of the cerebellar cortex is of clinical relevance, as abnormal serotonin metabolism has been observed in animal models and pathological cases of motor disorders involving the cerebellum, and as chronic intravenous administration of L-5-hydroxytryptophan (5 HTP), a precursor of serotonin, was the first treatment shown to improve significantly cerebellar symptoms. PMID- 11499401 TI - The pharmacology of mu analgesics: from patients to genes. AB - Morphine and most clinical opioids act through mu opioid receptors. Yet, their pharmacological profiles differ. The presence of incomplete cross-tolerance among these drugs clinically was one of the first indications that these mu opioids differed in their receptor mechanisms of action. This was followed by similar studies in preclinical models, which also found genetic differences in sensitivity toward morphine and other mu opioids. This concept of mu receptor multiplicity is now supported by antisense and gene knockout models. Although all the mu opioids are sensitive to antisense probes against the mu opioid receptor gene MOR-1, the sensitivity profiles of the drugs to the antisense probes differ based on the exon being targeted. Knockout mice also reveal striking differences. In one knockout mouse, morphine analgesia is completely lost while the potent mu drugs morphine-6beta-glucuronide and heroin both retain analgesic activity. Finally, cloning studies have identified at least seven different splice variants of the MOR-1 gene, with more likely. These studies illustrate the complexity of mu opioid pharmacology. PMID- 11499402 TI - Plasmalogens: workhorse lipids of membranes in normal and injured neurons and glia. AB - Plasmalogens are unique glycerophospholipids because they have an enol ether double bond at the sn-1 position of the glycerol backbone. They are found in all mammalian tissues, with ethanolamine plasmalogens 10-fold higher than choline plasmalogens except in muscles. The enol ether double bond at the sn-1 position makes plasmalogens more susceptible to oxidative stress than the corresponding ester-bonded glycerophospholipids. Plasmalogens are not only structural membrane components and a reservoir for second messengers but may also be involved in membrane fusion, ion transport, and cholesterol efflux. Plasmalogens may also act as antioxidants, thus protecting cells from oxidative stress. Receptor-mediated degradation of plasmalogens by plasmalogen-selective phospholipase A2 results in the generation of arachidonic acid, eicosanoids, and platelet activating factor. Low levels of these metabolites have trophic effects, but at high concentration they are cytotoxic and may be involved in allergic response, inflammation, and trauma. Levels of plasmalogens are decreased in several neurological disorders including Alzheimer's disease, ischemia, and spinal cord trauma. This may be due to the stimulation of plasmalogen-selective phospholipase A2. A deficiency of plasmalogens in peroxisomal disorders and Niemann-Pick type C disease indicates that this deficiency may be due to the decreased activity of plasmalogen synthesizing enzymes that occur in peroxisomes. PMID- 11499403 TI - The neurobiology of pain: developmental aspects. AB - Invasive procedures that would be painful in children and adults are frequently performed on infants admitted to the neonatal intensive care unit. This article discusses sensory responses to these procedures in the immature nervous system and highlights the fact that, in addition to causing distress and delayed recovery, pain in infancy is also a developmental issue. First, the immaturity of sensory processing within the newborn spinal cord leads to lower thresholds for excitation and sensitization, therefore potentially maximizing the central effects of these tissue-damaging inputs. Second, the plasticity of both peripheral and central sensory connections in the neonatal period means that early damage in infancy can lead to prolonged structural and functional alterations in pain pathways that can last into adult life. PMID- 11499404 TI - Sex hormones in experimental autoimmune encephalomyelitis: implications for multiple sclerosis. AB - For decades, it has been known that females are more susceptible than males to multiple sclerosis (MS). It has also long been appreciated that during late pregnancy there is a decrease in MS disease activity. Interestingly, these two observations have also been made in an extensively used animal model for MS, experimental autoimmune encephalomyelitis (EAE) in SJL mice. Female mice are more susceptible to disease than male mice, and there is an improvement in disease during late pregnancy. In this review, the role of sex hormones in each of these two observations is characterized in this EAE model using castration and exogenous hormone treatment strategies. The gender difference in EAE susceptibility is due primarily to a protective effect of testosterone in male mice. The decrease in disease severity during late pregnancy appears to be due at least in part to high levels of estriol, which characterize this time period. PMID- 11499405 TI - [27-year old male with AIDS and fever, lymphadenopathies, and immature cells in peripheral blood of recent appearance]. PMID- 11499406 TI - Genetic studies of spondylarthropathies. French Spondylarthropathy Genetic Study Group. AB - Influence of genetic factors in the spondylarthropathies has been clearly established. The goal of familial studies is to determine the respective weight of genetic factors and to discover new genes involved in the pathogenesis of spondylarthropathies. Among these factors, the HLA region appears to be of most importance. We are currently conducting a prospective multicenter French study aimed at collecting multiplex families of spondylarthropathies. We present preliminary results obtained from nearly 300 individuals corresponding to the first 20 collected families. Through the screening procedure, we have identified 57 patients of an average age of 42 and a mean age at onset of 23. The relative risk of spondylarthropathy in males as compared with females is 1.03. HLA typing revealed the presence of HLA-B27 in all affected patients from 19 of the 20 families, whereas in the last family, the two affected individuals shared the HLA B14 allele. Study of HLA class I allelic transmission confirmed a strong association of spondylarthropathy with HLA-B27 (p < 10(-9)). A weaker association was found with HLA-A28, HLA-C1 and HLA-C2, probably secondary to linkage disequilibrium between these alleles and HLA-B27. Study of HLA class II region indicated a possible protective effect of HLA-DR2 and an influence due to the presence of HLA-DR14. The risk to develop spondylarthropathy in HLA-B27+ relatives of patients decreased very rapidly in respect to the genetic distance, suggesting the genetic contribution of at least one other important gene distinct from the HLA region. These data encourage us to carry on a systematic screening of the whole human genome to discover implicated genes. PMID- 11499407 TI - Memory Clinics in Switzerland. Collaborative Group of Swiss Memory Clinics. AB - Currently, eleven Memory Clinics (MC) in Switzerland offer their services to patients, caregivers and family doctors. Their primary goal is the early diagnosis and management of dementia in elderly outpatients. Special emphasis is put on the detection of reversible causes of dementing disorders. Diagnoses and medical, pharmacological, neuropsychological and social treatment recommendations are established in a multidisciplinary consensus conference and communicated back to the referring physicians. The psychosocial therapies and supportive activities include counseling, memory training for patients, neuropsychological rehabilitation, caregiver groups, relaxation and biofeedback training, day care centers, Alzheimer's Tanzcafe, and special vacations for dementia patients. These activities vary from MC to MC according to the availability of resources. Research activities are an integral part of all MCs and range from the attempt to identify preclinical markers of Alzheimer's disease to studies of neuropathological correlates of cognitive disturbances. Moreover, patients are encouraged to participate in studies investigating new medications. MCs play an important role in the education and training of health care professionals and in raising awareness and support in the general population. PMID- 11499410 TI - [Social alienation. Lesson learned in clinical care of mentally ill patients at the Saint Anne Asylum. 1881]. PMID- 11499408 TI - Point mutations in the Plasmodium falciparum cg2 gene, polymorphism of the kappa repeat region, and their relationship with chloroquine resistance. AB - Based on the available DNA sequence data of the Plasmodium falciparum cg2 gene, we have hypothesized that 3 amino-acid substitutions, His275Gln, Gly281Ala, and His299Gln, may represent the key mutations that confer resistance to chloroquine. The presence of 14 tandemly repeated hexamer units in the kappa region has also been suggested to be indicative of chloroquine resistance. These 2 hypotheses were tested by determining the sequence of DNA fragments containing all 3 codons and kappa repetitive region (approximately 450-basepairs) for 53 randomly selected clinical isolates (obtained in Cameroon in 1994-97) with known response in vivo and/or in vitro to chloroquine. The cg2 genotypes based on the 3 codons and the response in vitro to chloroquine, as well as the number of kappa repeat units and responses in vivo and in vitro to chloroquine, were associated (P < 0.05). cg2 gene mutations were more common in parasites from patients with failure in vivo. However, this difference did not achieve statistical significance (P = 0.055). The sensitivity and specificity of the 3 codons and kappa repeat region to predict the response in vitro to chloroquine ranged between 75% and 85%. The sensitivity and specificity of these genetic markers to predict the response in vivo to chloroquine were of lower values. The kappa repeat region of the clinical isolates is polymorphic but characterized by several conserved features. PMID- 11499411 TI - [Transitory intoxicated delusions of alcoholic origin. 1906]. PMID- 11499412 TI - [Attempt at a physiological interpretation of paranoia and obsessive neurosis. 1935]. PMID- 11499414 TI - [Hallucinations. 1957]. PMID- 11499413 TI - ["Daseins-analysis" in psychiatry. 1951]. PMID- 11499415 TI - [Psychopathic personality. Attempt at a structural definition. 1967]. PMID- 11499416 TI - [Filial interpreters. 1910]. PMID- 11499417 TI - [Delusions of the imagination. 1911]. PMID- 11499418 TI - [Apropos of chronic hallucinatory psychosis]. PMID- 11499419 TI - [Origin and nature of hallucinations. 1922]. PMID- 11499420 TI - [Contribution to study of autistic thought and attitude (morbid rationalism). 1923]. PMID- 11499421 TI - [Psychopathological automatism is a result and not a cause. 1928]. PMID- 11499422 TI - [Cataleptic crises. Their diagnosis with pathological sleep. Their association with hysteria and catatonia. 1928]. PMID- 11499423 TI - [Video-laparoscopic surgery in urology]. PMID- 11499424 TI - Index of suspicion. Case 2. Diagnosis: Gastroesophageal reflux (GER). PMID- 11499425 TI - Index of suspicion. Case 3. Diagnosis: jaundice. PMID- 11499426 TI - What patient preparation techniques can be applied in a case of generalized anxiety disorder (GAD) to enhance compliance and improve outcome? PMID- 11499427 TI - How does one switch from one antidepressant to another when there has been a treatment failure? PMID- 11499428 TI - When should drug treatment be initiated in schizophrenia? PMID- 11499429 TI - Which medications should be prescribed to treat generalized anxiety disorder (GAD)?. PMID- 11499430 TI - How does one manage a patient with an anxiety disorder who has been self medicating with psychotropic drugs? PMID- 11499431 TI - What are the pharmacologic considerations when treating depression in patients with HIV infection? PMID- 11499432 TI - Prospecting in the pebble beds. PMID- 11499433 TI - Calibration of a 7.6 cm x 7.6 cm (3 inch x 3 inch) sodium iodide gamma ray spectrometer for air kerma rate. AB - An experimental procedure is described for converting a gamma ray spectral measurement from a 7.6 cm x 7.6 cm (3 inch x 3 inch) sodium iodide (NaI) detector to air kerma rate. The calibration procedure involves measuring the energy deposited in the detector using 10 radioactive sources of known activity covering an energy range from 60 keV to 1,836 keV. For each of the 10 sources, gamma ray spectra were measured with the source at different angles to the detector axis. The total energy deposited in the detector for the ten sources was confirmed by Monte Carlo calculations. The spectra measured at different angles were combined to produce a spectrum that would represent a homogeneous semi-infinite source of radiation. The resultant spectrum was then subdivided into 10 energy regions. Based on the known air kerma rates due to the sources, a calibration coefficient was calculated for each of the 10 energy regions. These calibration coefficients could then be used to convert the energy deposited in the 10 regions of an unknown spectrum to air kerma rate. The calibration procedure was confirmed by comparing the results from the detector with those from calibrated collimated beams of 137Cs and 60Co. A comparison of measurements using a calibrated pressurised ionisation chamber with those from a similar Nal spectrometer in Finland provided additional confirmation of the calibration procedure. PMID- 11499434 TI - Chromosome aberration analysis in radiotherapy patients and simulated partial body exposures: biological dosimetry for non-uniform exposures. AB - Chromosome aberration analysis was carried out in peripheral blood lymphocytes of cancer patients following radiotherapy of lungs, cervix and spine. Radiotherapy in the pelvic region involving large doses (6 Gy) showed an overdispersed distribution of dicentrics. However, when the doses were fractionated (three fractions of 2 Gy) distribution was found to be near Poisson. Spine irradiation covering almost all the lymphocytes pools, indicated a Poisson distribution. The data show that depending on the sites of exposure, the distribution of dicentrics in cells varies and hence there is a non-uniform distribution of lymphocytes in the body. The average dose to the lymphocytes was found to be one sixth of the partial body dose. Based on the non-Poisson distribution of aberrations, the fraction of lymphocytes irradiated, mean dose to the fraction and part of the body exposed was calculated in a case of acute 6 Gy pelvic irradiation. The fraction of cells irradiated was calculated to be 4.11% and the portion of the body exposed was approximately 16.8%. The dose to the irradiated fraction was found to be 5.4 Gy, which is in agreement with the given dose of 6 Gy. In simulated exposures the u values increased systematically with the decrease in fraction of irradiated cells and the calculated dose to the fraction was also in good agreement with the true dose. PMID- 11499435 TI - Radiation exposure to patients during paediatric cardiac catheterisation. AB - Radiation doses were investigated for 18 infants and children undergoing cardiac catheterisation procedures with thermoluminescence dosemeters. The range of integrated current values used during cardiac catheterisation procedure was wide, from 12.2 to 1195.8 mA.min (mean 604.9). The average was 22.9 mA.min for fluoroscopy, and 616.1 mA.min for cineangiography, and the ratio of cineangiography to fluoroscopy ranged from 10.5 to 89.5 with an average of 34.0. The cineangiographic contribution was estimated to be 90% of the total doses. The entrance surface doses and thyroid doses varied widely. The ratio of maximum to minimum for entrance surface doses was 98.5, for left and right thyroid it was 59.8 and 104.4, respectively. The analysis of the entrance surface doses in three age groups showed that there was no significant difference among them. There was a weak inverse relation between the thyroid dose and the weight of the patient, while no correlation was found between the thyroid dose and the entrance surface dose. The average of entrance surface doses to the patients was 847.3 mGy, which was 1-2 orders of magnitude higher than common X ray examinations. PMID- 11499436 TI - Evaluation of environmental dose at JCO using luminescence from quartz stimulated by blue light. AB - The environmental dose due to the recent nuclear accident at JCO, Japan, was estimated using luminescence optically stimulated from unheated quartz. Two methods originally developed for dating analysis, the single aliquot additive dose method and the single aliquot regeneration added dose method, were employed to confirm the dose rate. Consistent results were obtained from both methods and from thermoluminescence measurements. Although the dose rate values had lower precision than can be obtained from heated materials, it is suggested that luminescence from sedimentary quartz can usefully be employed in retrosepective dosimetry. PMID- 11499437 TI - Calibration curves for biological dosimetry by fluorescence in situ hybridisation. AB - Dose-response curves were measured for the induction of chromosomal aberrations in peripheral blood lymphocytes after acute exposure in vitro to 60Co gamma rays. Blood was obtained from four different healthy donors, and chromosomes were either observed at metaphase, following colcemid accumulation, or prematurely condensed by calyculin A. Cells were analysed in three different Italian laboratories. Chromosomes 1, 2, and 4 were painted, and simple-type interchanges between painted and non-painted chromosomes were scored in cells exposed in the dose range 0.1-3.0 Gy. The chemical-induced premature chromosome condensation method was also used combined with chromosome painting (chromosome 4 only) to determine calibration curves for high dose exposures (up to 20 Gy X rays). Calibration curves described in this paper will be used in our laboratories for biological dosimetry by fluorescence in situ hybridisation. PMID- 11499438 TI - Bayesian prior probability distributions for internal dosimetry. AB - The problem of choosing a prior distribution for the Bayesian interpretation of measurements (specifically internal dosimetry measurements) is considered using a theoretical analysis and by examining historical tritium and plutonium urine bioassay data from Los Alamos. Two models for the prior probability distribution are proposed: (1) the log-normal distribution, when there is some additional information to determine the scale of the true result, and (2) the 'alpha' distribution (a simplified variant of the gamma distribution) when there is not. These models have been incorporated into version 3 of the Bayesian internal dosimetry code in use at Los Alamos (downloadable from our web site). Plutonium internal dosimetry at Los Alamos is now being done using prior probability distribution parameters determined self-consistently from population averages of Los Alamos data. PMID- 11499439 TI - Determination of neutralisation rates of 218Po ions in air. AB - The electric charge of the radon decay product, 218Po, has an influence on its mobility characterised by the diffusion coefficient, which chiefly controls the formation of the radioactive aerosol by attachment and the plateout processes on surfaces. These processes strongly affect the dose relevant quantities like concentration and particle size distribution of the short-lived radon decay products. The neutralisation rate of the positive 218Po ions in environmental air was determined quantitatively in chamber experiments. The experimental results show the great influence of the ionisation rate and the humidity concentration on the neutralisation rate of the 218Po ions in air. The obtained neutralisation rates were described by equations, taking into account the neutralisation processes like ion recombination, charge transfer and electron scavenging. In 'normal' environmental air with an ionisation rate of 45 muR.h(-1) = 1.16 x 10( 8) C/(kg.h) and a relative humidity of 50% (T = 20 degrees C), a neutralisation rate of the 218Po ions of 1.54 x 10(-2) s(-1) was obtained. Taking into account this neutralisation rate leads to a fraction of about 51% of the Po cluster that are neutral in air. PMID- 11499440 TI - Comparison of two JAERI phantoms and the problems discovered. AB - During the course of an intercomparison exercise it was possible to compare two JAERI phantoms with each other by using a multi-energy photon emitting lung set (241Am/152Eu). One belonged to the IAEA (Vienna), the other belonged to the Human Monitoring Laboratory (Ottawa). The intercomparison of the phantoms showed that they were statistically distinct from each other, although the differences were small. The counting efficiencies varied from each other by about 4% at 17 keV and 2% at photon energies above 17 keV. It was concluded that these difference were either due to small variations of chest wall thickness during the manufacturing process or positioning errors. The intercomparison also revealed a serious problem with one of the overlay plates of the HML's phantom. The adipose mass fraction of the overlay plate was found to be much greater (approximately 40%) than the manufacturer's stated value (10%). PMID- 11499441 TI - Physical parameters and dose factors of the radon and thoron decay products. AB - The dose per exposure unit of the short-lived radon and thoron decay products was calculated using a dosimetric approach. The calculations are based on a lung dose model with the structure that is related to the ICRP 66 respiratory tract model. The dose relevant parameters, unattached fraction of the decay product clusters (fp) and size distribution of the unattached and aerosol-attached decay products for different living and working places are reported. Taking into account these characteristics the dose conversion factors (DCF) of the radon and thoron decay products were estimated. In addition, the living and working places were divided concerning their aerosol parameters like particle number concentration and activity size distribution. PMID- 11499442 TI - Outdoor background ELF magnetic fields in an urban environment. AB - Classification of 'exposed/non-exposed' subjects in epidemiological studies concerning the possible cancer risk associated with ELF magnetic field exposure is based on the a priori assumption of magnetic field value cut-off points that, often, are defined equal to minimum exposure levels typical of a population residing near high voltage facilities (0.1-4.2 microT), but in some cases an environmental magnetic field level not associated with transmission lines can exist. The results of an ELF magnetic field survey in an Italian urban area (about 1 million inhabitants) are presented: average field levels are correlated with population density of different districts. Exposure indexes are obtained, which are compared with those evaluated in studies regarding domestic exposure: background average levels result in comparable to cut-off points in epidemiological studies, but in some districts with high population density, they are much higher. This shows that knowledge of background magnetic field level in urban areas can assume a significant role in exposure assessment. PMID- 11499443 TI - The potential for reduction of radiation doses to patients undergoing some common X ray examinations in Tanzania. AB - The potential for patient dose reduction in diagnostic radiology was investigated in five major Tanzanian hospitals. The aim of this study was to follow up previously reported suggestions for improved practices to achieve dose reductions. The suggestions were implemented and entrance surface dose measurement repeated by using well-calibrated LiF thermoluminescence dosemeters. The results show that dose reductions in chest PA X ray examinations ranged from 15% to 50%. For abdomen AP and pelvis AP X ray examinations, the dose reductions ranged from 24% to 73% and from 25% to 72%, respectively. The respective dose reductions for lumbar spine AP and LAT projections ranged from 4% to 58% and from 16% to 77%. Interestingly, the majority of radiographs obtained after the implementation of dose reduction measures were useful for intended diagnosis according to the opinion of radiologists. It is concluded that significant dose reductions can be achieved in the country without loss of diagnostic information. Such dose reductions also predict reductions of radiation risk to patients. PMID- 11499444 TI - Susceptibility of calcium-deficient hydroxyapatite-collagen composite to irradiation. AB - The ESR spectrum of calcium-deficient hydroxyapatite (DAp) has the peaks assigned to the hyperfine structure of dangling H+ (hyperfine coupling constant = 50.8 mT, I = 1/2) due to the HPO(4)2- ion which captured the hole released by X ray irradiation. This ESR peak intensity in DAp with collagen (c-DAp) decreased in an increase in the amount of collagen added into Ca(H2PO4)2H2O (MCP) electrolytic solution, because dangling H+ of HPO(4)2- binds to the carboxyl group of collagen due to the hydrogen bond. The ESR signal intensities at near g = 2 in DAp, c-DAp, and stoichiometric hydroxyapatite (HAp) after X ray irradiation, were proportional to the absorbed dose in the range from 6 to 380 Gy. These ESR signal intensities decreased when the X ray-irradiated DAp, c-DAp, and HAp was suspended in the simulated body fluid. This fact suggests that the surface layer contained high density of ESR active species in DAp, c-DAp, and HAp dissolved in the simulated body fluid. Therefore, with the dosemeter utilising such biomaterials as tooth and bone, sufficient care must be paid to the effect of body fluid. PMID- 11499445 TI - Blending incremental and stratified layering techniques to produce an esthetic posterior composite resin restoration with a predictable prognosis. AB - Composite resin restorations play an ever-increasing role as routine restorations in everyday clinical practice. However, the long-term prognosis of these restorations is still widely debated and open to question. The restorative protocols are still evolving, whether for direct or indirect placement, and little evidence is available in the scientific literature as to the ideal choice of site, technique, and category for placement. This article discusses the problems encountered and suggests a clinical restorative protocol to optimize composite resin placement. PMID- 11499446 TI - Simple technique to fabricate provisional restorations for porcelain veneers. AB - Fabrication of provisional restorations for teeth prepared to receive porcelain veneers can be a time-consuming and difficult task. This article describes a simple, quick, and predictable procedure to make these restorations. A clinical case is presented to illustrate the technique. PMID- 11499447 TI - Effect of light power density on development of elastic modulus of a model light activated composite during polymerization. AB - PURPOSE: Elastic modulus development during polymerization of a composite is a measure of the polymerization maturity and the restoration's ability to transfer stress to enamel and dentin. The characteristics of elastic modulus development in real time during cure are largely unknown. The purpose of this study was to evaluate the effect of light power density and total energy density on the early development of elastic modulus for a light-activated composite. METHODS: Cylindrical specimens of a model hybrid composite were tested in flexure in a dynamic mechanical analyzer (DMA). Specimens were light-activated (Variable Intensity Polymerizer, Bisco, Itasca, Illinois) for 60 seconds. Elastic modulus was measured continuously for 5 minutes from the start of light activation. Development of elastic modulus was assessed for six different light power densities and two reduced power density levels given at longer exposure duration to provide similar energy density values. One-way analysis of variance with Tukey's post hoc comparison test was used to evaluate significant differences of elastic modulus at p = .05. RESULTS: The rates of elastic modulus development and final moduli were dependent on the light power density applied. Composite specimens cured by equivalent energy densities using short times and high power density or long times and low power density produced equivalent elastic moduli. Elastic moduli for emitted power densities between 400 and 600 mW/cm2 (160-260 mW/cm2 measured at the specimen surface) were not significantly different (p > .05). CONCLUSIONS: Light power densities greater than 160 mW/cm2 measured at the specimen surface resulted in elastic moduli that were not significantly different. Equivalent energy densities produced comparable elastic moduli. PMID- 11499448 TI - Nightguard vital bleaching and its effect on enamel surface morphology. AB - BACKGROUND: The effect that nightguard vital bleaching (NGVB) has on enamel surface morphology is a subject of debate. Previous studies that have evaluated the effect of NGVB on the enamel surface report minimal changes to changes that appear to worsen post-treatment. The purpose of this in vivo NGVB study was to evaluate by scanning electron microscopy (SEM) the effects that 10% carbamide peroxide has on enamel morphology after 2 weeks of treatment and at 6 months post treatment. MATERIALS AND METHODS: Ten patients participated in the study. Each participant wore a guard filled with an active whitening solution for 8 to 10 hours per day for 14 treatment days. An impression of the study teeth (maxillary incisors) was taken at baseline, after 14 days of treatment, and at 6 months post treatment, and an epoxy cast made. The epoxy casts were prepared for viewing under the SEM and photographs were taken at 200 times and 2,000 times magnification. Six examiners evaluated changes in enamel surface morphology by comparing the SEM photographs taken at baseline, on treatment day 14, and at 6 months post-treatment. Still masked, the examiners also compared each patient's baseline:treatment day 14 and baseline:6 months post-treatment photographs with photographs of a known standard. RESULTS: This in vivo study demonstrated that a 14-day regimen of NGVB using a 10% carbamide peroxide solution had minimal effect on the surface morphology of enamel and that the effects did not worsen over time. PMID- 11499449 TI - Talking with patients. Posterior composites. PMID- 11499450 TI - Ask the experts. Porcelain veneers. PMID- 11499451 TI - Posterior resin-based composite restorations: clinical recommendations for optimal success. AB - Resin-based composites are increasingly used for the restoration of defects in posterior teeth. This review describes, illustrates and discusses important clinical aspects of the posterior composite technique. A relatively new stratification concept oriented to the development of functional and anatomic restorations is proposed. PMID- 11499452 TI - Advancing scientific discourse in the controversy surrounding the Comprehensive System for the Rorschach: a rejoinder to Meyer (2000). AB - A recent commentary by Meyer (2000) in the Journal of Personality Assessment alleged that Rorschach critic Wood and his colleagues had intentionally published information that they knew to be in error. To substantiate this contention, Meyer's commentary published information that was part of the peer review process at another journal. In this rejoinder, we present factual information that shows we have consistently acted in good faith. This rejoinder suggests that the scientific debate regarding the Comprehensive System for the Rorschach is unlikely to be advanced by speculating about the intentions of Rorschach critics, or by publishing information from the peer review process that is usually kept confidential. PMID- 11499453 TI - The Sexual Compulsivity Scale: further development and use with HIV-positive persons. AB - We examined the reliability and validity of the Sexual Compulsivity Scale for use in assessing HIV-positive men and women. Measures collected from 287 men and women recruited from the community, the majority of whom were African American and over the age of 30, showed that the Sexual Compulsivity Scale was reliable for men (alpha = .89) and women (alpha = .92). Correlations with measures of sexual behaviors and numbers of sex partners supported the concurrent validity of the Sexual Compulsivity Scale for men and women. Results of discriminant function analyses that included participant age, use of nonalcohol drugs, the Sexual Compulsivity Scale, and scores on measures of trait anxiety, obsessive compulsiveness, future pessimism, cognitive depression, and borderline personality characteristics showed that sexual compulsivity clearly discriminated between persons with 1 or no sex partners and persons with multiple partners in the past 3 months, for both men and women, suggesting evidence for criterion related validity. Associations between sexual compulsivity scores and other markers of psychopathology showed different patterns for men and women; sexual compulsivity was associated with indexes of psychopathology in men but not in women. We conclude that the Sexual Compulsivity Scale is reliable and valid in assessing men and women infected with HIV, although sexual compulsivity may present differently between genders in relation to other forms of psychopathology. PMID- 11499454 TI - Symptom endorsement differences on the Children's Depression Inventory with children and adolescents on an inpatient unit. AB - Responses to the Children's Depression Inventory (CDI; Kovacs, 1992), administered during intake, were collected from 521 children and adolescents (aged 7 to 17) at an inpatient crisis stabilization unit. Participants were grouped into 1 of 3 diagnostic groups: solely depressive, solely aggressive, or both depressive and aggressive. Self-report of symptoms for each diagnostic group, age and gender differences, and racial and ethnic differences in symptoms were examined in this study. There was a significant difference between the endorsement pattern of solely depressive and solely aggressive participants, whereas those categorized as both depressive and aggressive displayed an endorsement pattern similar to those who were solely aggressive. There was a significant gender difference in overall symptom report, with girls showing higher overall symptom levels than boys. This gender difference was significant for both the younger and the older age groups. These results held true even when gender was covaried out of the diagnostic group analyses and when diagnostic group was covaried out of the gender analyses. Symptom endorsement did not differ based on race and ethnicity. The primary contribution of this study centers around the findings from the item analyses of the CDI. These results are discussed in relation to the discriminant validity of the CDI and the need for additional research into comorbidity. PMID- 11499455 TI - Research Validity Scales for the NEO-PI-R: additional evidence for reliability and validity. AB - We examined the reliability and validity of the research validity scales (Schinka, Kinder, & Kremer, 1997) for the NEO-Personality Inventory-Revised (NEO PI-R) in a clinical sample. The Negative Presentation Management (NPM) and Positive Presentation Management (PPM) scales were found to have satisfactory internal consistency reliability. Support for the validity of these scales was provided by the pattern of convergent and discriminant correlations with respective Personality Assessment Inventory (PAI) validity scales. Finally, PAI profiles of individuals with invalid NPM scores were found to differ significantly from those with valid NPM scores. Comparisons of the invalid profiles with profiles from other clinical samples provided additional support for the use of the NPM scale as a measure of negative impression management. PMID- 11499456 TI - A model for brief assessment of attachment and its application to women in inpatient treatment for trauma-related psychiatric disorders. AB - We adapted self-report measures of attachment style to the psychological assessment of women in specialized inpatient treatment for trauma-related disorders. The study employed 2 measures of adult attachment style, the Relationship Questionnaire (Bartholomew & Horowitz, 1991) and the Adult Attachment Scale (Collins & Read, 1990) as well as our Current Attachment Relationships questionnaire, which assesses the extent of social support in secure attachments. We administered these measures to 99 patients and to a convenience sample of 154 women in the community. We found modest correspondence between the 2 attachment style measures and substantial relations between attachment styles and range of secure attachment relationships. Women in the trauma sample reported insecure attachment styles and relatively few secure attachment figures. We discuss the implications of these findings for clinical assessment. PMID- 11499457 TI - The Rotter Incomplete Sentences Blank: examining potential race differences. AB - The Rotter Incomplete Sentences Blank (RISB; Rotter, Lah, & Rafferty, 1992; Rotter & Rafferty, 1950) was administered to African American (n = 94) and White (n = 100) members of fraternal organizations at a large Midwestern university to explore possible influences of racial group and socially desirable response tendencies. The RISB protocols were reliably scored (interrater agreement = 92%) by 3 independent scorers blind to participant racial membership. The mean scores of the 2 racial groups were not significantly different (p > .05). However, using the traditional categorical placement scores resulted in a disproportionate assignment rate for African American scores to the maladjusted group than would be anticipated by mere chance. PMID- 11499458 TI - Rorschach administration: a comparison of the effect of two instructions. AB - The effect of administering the Rorschach Inkblot Method under 2 instructional sets was compared across 19 outcome measures. Sixty healthy participants randomly received either the short instruction "What might this be?" originally developed by Rorschach (1942) and recommended in the Comprehensive System (Exner, 1993), or a long instruction (Killingmo, 1980), which for many years has been the standard instruction in Norway. The short instruction produced significantly more questions to the examiner about the test and more brief protocols R < 14) than did the long one. However, for the traditional summary Rorschach scores no between-group differences were observed for the 2 instructional sets. It is suggested that if future studies of larger clinical and more representative samples demonstrate corresponding results, a more elaborate standard instruction might be preferable. PMID- 11499459 TI - Readministration of the MMPI-2 following defensive invalidation in a military job applicant sample. AB - Job applicants whose response style on the Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer) is excessively defensive create a dilemma for psychologists working in personnel selection settings. Clinical guidelines recommend the profile be considered uninterpretable, but the absence of usable testing data could lead to the elimination of otherwise qualified candidates. Readministering the MMPI-2 with altered instructions to reduce defensive responding has been suggested as an alternative. This option has lacked empirical support until recently. One study (Butcher, Morfitt, Rouse, & Holden, 1997) evaluated the effects of altered instructions on retest validity in a civilian job applicant sample, with the majority of participants obtaining valid and normal profiles on retest. The purpose of this study was to determine if these results would be replicated in a military job applicant sample. Participants were 97 military personnel who completed the MMPI-2 as part of application for selection to instructor duty. Forty-seven participants had obtained invalid profiles and retook the MMPI-2 after receiving instructions intended to reduce defensiveness. The results showed that 83% of retested participants obtained valid profiles on the second MMPI-2. The second test results were very similar in profile to those obtained from a comparison group of 50 participants whose initial MMPI-2 results were valid. The findings are discussed in terms of study limitations and future research directions. PMID- 11499460 TI - Rorschach protocols from children and adolescents with Asperger's disorder. AB - Rorschach protocols from 24 boys with Asperger's Disorder matched by age to 24 boys with other emotional or behavioral disorders (the contrast group) were compared to each other and to Exner' s (1995) normative data. Eight variables based on Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) criteria and a review of the literature for Asperger's Disorder were predicted to discriminate between groups with the Asperger's group having more extreme scores. Five variables (COP, CDI, H, M, and EA) were significantly different from the contrast group and T and WSumC were significantly different from the normative data in both the Asperger's group and the contrast group. PMID- 11499461 TI - Effectiveness of NEO-PI-R research validity scales for discriminating analog malingering and genuine psychopathology. AB - We investigated the research validity scales for the NEO Personality Inventory Revised (NEO-PI-R) proposed by Schinka, Kinder, and Kremer (1997): Positive Presentation Management (PPM) and Negative Presentation Management (NPM). Additionally, an experimental analog to the Minnesota Multiphasic Personality Inventory-2's (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) F-K index was calculated by subtracting the raw score on PPM from the raw score on NPM (NPM-PPM). In 2 studies, all indexes showed significant between-group differences when samples of analog malingerers (n = 97) were contrasted with psychiatric outpatients (n = 272). The sensitivity and specificity of these validity indexes indicated that although none performed well in extremely low base rate environments, the NPM and NPM-PPM indexes showed promise when the base rate of faking bad rose to higher levels. PMID- 11499462 TI - Clinical identification of compensatory structures on projective tests: a self psychological approach. AB - In this article I discuss compensatory structure, a concept from Kohut's (1971, 1977) psychology of the self that is not as familiar as Kohut's other views about the self. Compensatory structures are attempts to repair selfobject failure, usually by strengthening idealization or twinship in the face of mirroring deficits. Compensatory structures, particularly their early indications, can be detected on projective tests for identifying adaptive resources and treatment potential. The clinical identification of compensatory structures on test findings is described using Rorschach and Thematic Apperception Test (Murray, 1943) content. Particular attention is devoted to the 2-part process of demonstrating first, an injury to the self, and second, how attempts to recover from such injuries can be detected on projective tests. Clinical examples are provided, and the differentiation between compensatory structures and defenses and sublimation is discussed. PMID- 11499463 TI - Distinguishing narcissism and hostility: similarities and differences in interpersonal circumplex and five-factor correlates. AB - Narcissism and hostility are both characterized by dysfunctional social interactions, including tendencies to perceive slights, experience anger, and behave aggressively. The aim of this study was to examine the similarities and differences of narcissism and hostility, using 2 conceptual tools-the interpersonal circumplex and the Five-factor model. In a sample of 292 undergraduate men and women, composite measures of hostility (i.e., Buss-Perry Aggression Questionnaire [Buss & Perry, 1992] and Cook-Medley Hostility [Cook & Medley, 1954] total scores) were inversely correlated with affiliation and unrelated to dominance. In contrast, composite narcissism scores (i.e., Narcissistic Personality Inventory) were positively correlated with dominance and inversely correlated with affiliation. Examination of components of these traits revealed additional similarities and differences, as did associations with other dimensions of the Five-factor model. These findings suggest that the traits of narcissism and hostility are distinguishable by their interpersonal referents, as are their components. PMID- 11499464 TI - Finger reconstruction with a free neurovascular wrap-around flap from the big toe. AB - From 1983 to 1998, 16 cases of finger reconstruction with a free neurovascular wrap-around flap from the big toe were treated. Fourteen cases were successful, and two cases failed. The authors reviewed these cases on the average of about 38 months after surgery. Pinch power was 51 percent of the unaffected normal hand, and two-point discrimination was 7.6 mm. The mean resorption of the grafted bone was 13 percent in width and 9 percent in length. There were no complications such as fracture of the grafted bone, nonunion, and pulp dislodgement. This procedure provided length, stability, and adequate sensibility for a functional pinch and grasp. Sensory return to the wrap-around flap on the thumb was often greater than for the same area on the opposite foot. The donor site of the wrap-around flap was acceptable, both aesthetically and functionally, and allowed the wearing of open-toed shoes by young women. Finger reconstruction with a wrap-around flap from the big toe yielded excellent cosmetic and functional results in cases involving amputation at the level of the metacarpophalangealjoints or distal to it. In addition, this procedure was an excellent choice for treatment in cases involving avulsion injuries of the fingers and reconstruction of soft-tissue defects after tumor excision. PMID- 11499465 TI - Reconstruction after extirpation of the auricle. AB - The purpose of this study is to introduce the procedure and results of a combined free flap reconstruction with an external ear prosthesis after tumor extirpation from the lateral side of the head. Over the last 3 years, four patients have undergone total auricle extirpation for varying tumor processes. Each was reconstructed with a radial forearm flap, preserving the external auditory meatus. After primary healing and adjuvant therapy, a prosthetic pinna was applied. Combined use of microsurgical techniques with a prosthetic ear has produced an aesthetically superior result for large soft-tissue defects of the side of the head. This combined technique is able to provide the best aspects of both microsurgical and prosthetic reconstructions for a superior result for the patient. PMID- 11499466 TI - A secure locking knot for microsuturing. AB - Perfect balance between maximal suture strength and minimal foreign-body reactivity guarantees success, using microsurgical techniques. The proposed initial locking knot allows optimal edge approximation. It has strength, it is simple to master, and is not too bulky. The knot is kept in its first position, without danger of slipping, before securing it with an additional knot of two loops. No additional knots are necessary, because the completed knot may fail by breaking, but never by slippage. PMID- 11499467 TI - Use of a mechanical leech in a reverse-flow radial forearm flap: case report. AB - The reverse-flow radial forearm flap provides excellent coverage for distal upper limb defects. It is simply raised and does not require microsurgical skills. However, since its vascular pedicle is reversed, its venous outflow can be significantly diminished because of the venous valves. The authors present the case of a 16-year-old patient with a sagittal amputation of the radial aspect of the right thumb, who manifested at the time of surgery marked venous engorgement of a reverse-flow radial forearm flap. This was successfully relieved by the placement of a mechanical leech consisting of a Silastic rubber catheter--of the kind used to gain central vascular access in newborns--introduced in the lumen of the reversed vein at the extremity of the flap. This permitted intermittent evacuation of blood from the flap postoperatively, contributing to the success of this procedure. The technique used is detailed and pertinent literature is reviewed. PMID- 11499468 TI - Clinical classification of free-flap perfusion complications. AB - Free-tissue transfer is a standard procedure in hand and other plastic and reconstructive surgery. The aim of the present article was to present a new clinical classification of disturbances of the circulation, and to assess the results of treating the specific complication at each of the stages described. Within a period of 29 months, 194 free microvascular flaps were prepared and evaluated in a prospective study. Postoperative monitoring was carried out from a purely clinical point of view. A distinction was made between arterial and venous circulatory disturbances and, in both cases, four stages were characterized, according to the time required for recapillarization, the color of the transfer, and bleeding on puncture. Complications occurred in 69 cases (36 percent), including intraoperative revisions during a first operation. The ratio of arterial to venous insufficiency was 33:35. In the presence of arterial disturbances of the circulation, the proportion of losses increased in progressive stages to 41 percent. With venous disturbances, losses which occurred in stages 1 and 2 were predominantly partial. In 164 cases (85 percent), the flaps remained undamaged. In 15 cases (8 percent), there was partial loss of the transfer and, in a further 13 cases (6 percent), there was complete loss. The authors' results confirmed that the presented clinical classification is, indeed, a measurement of the severity of circulatory impairment. Despite the relatively high complication rate, the use of various methods of treatment finally led, in the overwhelming majority of cases, to a positive result. PMID- 11499469 TI - The fate of neurotization techniques on reinnervation after denervation of the gastrocnemius muscle: an experimental study. AB - In nerve injuries, if it is not possible to reinnervate muscle by using neurorrhaphy and nerve grafting technique, reinnervation should be provided by the use of neuroization-directly implanting motor nerve into muscle. A comparative study of three techniques of neurotization is presented in rabbits. In this experimental study, a total of 40 white New Zealand rabbits were used and divided into four groups, each including 10 rabbits. In the first group (control- Group 1), only surgical exposure of the gastrocnemius muscle, main muscle nerve (tibial nerve), and peroneal nerve was done, without any injury to the nerves. In the second group (direct neurotization group--Group 2), the tibial nerve was transected, and the peroneal nerve, which had already been divided into fascicles, was implanted into the lateral head of the gastrocnemius muscle aneural zone. In the third group (dual neurotization group--Group 3), the tibial nerve which had been transected and re-anastomosed, and the peroneal nerve were implanted into the lateral head of the gastrocnemius muscle. In the last experimental group (hyperneurotization group--Group 4), fascicles of the peroneal nerve were implanted into the lateral head of the gastrocnemius, preserving the tibial nerve. Six months later, changes in the histologic pattern and the functional recovery of the gastrocnemius muscle were investigated. It was found that functional recovery was achieved in all neurotization groups. Groups with the tibial nerve transected had less muscular weights than those of groups with the tibial nerve intact. EMG recordings showed that polyphasic and late potentials were frequently seen in groups with the tibial nerve transected. Degeneration and regeneration of myofibrils was observed in such groups as well. New motor end-plates, including vesicles, were formed in a scattered manner in all neurotization groups. As a result, the authors conclude that direct and dual neurotization techniques are useful in peripheral nerve injuries, if it is not possible to reinnervate muscle by using neurorraphy and nerve grafting, and that there is no suggested superiority among these techniques. PMID- 11499470 TI - Enhanced reinnervation of the paralyzed orbicularis oculi muscle after insulin like growth factor-I (IGF-I) delivery to a nerve graft. AB - Facial paralysis (FP) remains today one of the most disturbing cranial nerve disorders. The present study utilized the rat model of FP and examined a dual approach of combining the current microsurgical treatment of cross-facial nerve graft (CFNG) with local administration of insulin-like growth factor-I (IGF-I). The efficacy of this combined treatment approach was assessed by motor end-plate analysis of the reinnervated orbicularis oculi muscle (OOM). Local administration of IGF-I (50 microg/ml) to the CFNG demonstrated a 61 percent increase in the number of end-plates in the reinnervated OOMs, compared to the OOMs reinnervated with CFNG plus vehicle. These results indicate that the local therapeutic augmentation of IGF-I levels at the coaptation site(s) of the CFNG may, in fact, enhance reinnervation of muscle and recovery of function in general. PMID- 11499471 TI - Nerve conduit using fascia-wrapped fibrocollagenous tube. AB - The purpose of this study was to evaluate the application of autogenous fascia as a framework of a fibrocollagen tube for a nerve conduit in Japanese white rabbits. The fascia was wrapped with a double layer around a silicone rod 3.0 mm in diameter. After the implantation of the fascia-wrapped silicone rod into a subcutaneous pocket for 2 weeks, a 3.0 x 30-mm fascia-wrapped fibrocollagen (FFC) tube was prepared. With microvascular techniques, the tube was interposed into a right sciatic nerve gap which was 25 mm long. The results of nerve regeneration in the FFC tube group, in particular, the formation of epineurium, were able to stand comparison with the results of a control group under both histologic and electron micrographic examination. PMID- 11499472 TI - Changes in blood flow in the partially elevated epigastric pedicled flap in response to occlusion of the femoral artery in rats. AB - In this study, the authors used a custom-designed clamping method and laser Doppler flowmetry to investigate the short-term autoregulatory patterns of blood flow (BF) in partially elevated pedicled epigastric flaps in rats (n=11). The femoral artery was clamped for 30, 60, 120 and 180 sec directly after flap elevation and 40 to 80 min after flap elevation. Changes during and after clamping indicate the functioning of autoregulatory factors in the vessels of the epigastric partially elevated flap. The longer the clamp ischemia is in a flap, the greater (p < 0.0001) is the need for a compensatory increase in BF and for a longer time (p < 0.0001) in the flap after clamp release. They compared the responses of BF directly after flap elevation with those 40 to 80 min after flap elevation. The results suggest a significant increase in the autoregulatory capacity of the pedicled partially elevated flap in measurements performed 40 to 80 min after flap elevation (p = 0.043). They conclude that the novel clamping method seems to be reliable when studying the short-term autoregulatory patterns of the epigastric partially elevated pedicled flap. PMID- 11499473 TI - Methodology for the development of a drug library based upon collision-induced fragmentation for the identification of toxicologically relevant drugs in plasma samples. AB - The possibility of creating a robust mass spectral library with use of high performance liquid chromatography-atmospheric pressure-electrospray ionization (HPLC-AP-ESI) for the identification of drugs misused in cases of clinical toxicology has been examined. Factors reported as influencing the fragmentation induced by "source transport region collision induced dissociation" (CID) have been tested in this study (i.e. solvent, pH, different acids or buffer salts and their concentration, different organic modifiers and the modifier concentration). The tests performed on a few "model drugs" were analysed with use of two different single quadrupole instruments. The large number of mass spectra obtained appears to be affected by the mobile phase conditions to only a minor extent. This also holds for the mass spectra obtained at two different instruments (laboratories). Subsequently breakdown curves have been measured for about 20 randomly chosen drugs by variation of the kinetic energy of their ions in the CID zone through changing the fragmenter voltage. These breakdown curves were used to optimize the fragmenter voltage for each drug. The optimized fragmenter voltages were then applied by use of a variably ramped fragmenter voltage to acquire mass spectra for the library. The chromatographic separations were run on a Zorbax Stable bond column using a 10-mM ammonium formate acetonitrile gradient method. Spiked blank serum and patient samples with a total of 40 different drugs were extracted with use of a standard basic liquid-liquid extraction (LLE) method. A search of significant peaks in the chromatogram by application of the developed mass spectral library is shown to result in a more than 95% positive identification. reserved. PMID- 11499474 TI - Use of novel solid-phase extraction sorbent materials for high-performance liquid chromatography quantitation of caffeine metabolism products methylxanthines and methyluric acids in samples of biological origin. AB - An automated reversed-phase high-performance liquid chromatographic (RP-HPLC) method, using a linear gradient elution, is described for the simultaneous analysis of caffeine and metabolites according to their elution order: 7 methyluric acid, 1-methyluric acid, 7-methylxanthine, 3-methylxanthine, 1 methylxanthine, 1,3-dimethyluric acid, theobromine, 1,7-dimethyluric acid, paraxanthine and theophylline. The analytical column, an MZ Kromasil C4, 250 x 4 mm, 5 microm, was operated at ambient temperature with back pressure values of 80 110 kg/cm2. The mobile phase consisted of an acetate buffer (pH 3.5)-methanol (97:3, v/v) changing to 80:20 v/v in 20 min time, delivered at a flow-rate of 1 ml/min. Paracetamol was used as internal standard at a concentration of 6.18 ng/microl. Detection was performed with a variable wavelength UV-visible detector at 275 nm, resulting in detection limits of 0.3 ng per 10-microl injection, while linearity held up to 8 ng/microl for most of analytes, except for paraxanthine and theophylline, for which it was 12 ng/microl and for caffeine for which it was 20 ng/microl. The statistical evaluation of the method was examined performing intra-day (n=6) and inter-day calibration (n=7) and was found to be satisfactory, with high accuracy and precision results. High extraction recoveries from biological matrices: blood serum and urine ranging from 84.6 to 103.0%, were achieved using Nexus SPE cartridges with hydrophilic and lipophilic properties and methanol-acetate buffer (pH 3.5) (50:50, v/v) as eluent, requiring small volumes, 40 microl of blood serum and 100 microl of urine. PMID- 11499475 TI - On-line identification of 3,4-methylenedioxymethamphetamine in human urine by non aqueous capillary electrophoresis-fluorescence spectroscopy at 77 K. AB - The analytical profiles for 3,4-methylenedioxymethamphetamine (3,4-MDMA) and related amphetamines in urine samples are described for non-aqueous capillary electrophoresis-fluorescence spectroscopy. 3,4-MDMA was detected and identified on-line, using a cryogenic molecular fluorescence technique at 77 K. Under optimized conditions, baseline separation of the selected compounds was achieved in less than 12 min. Precision was evaluated by measuring the repeatability and intermediate precision of the migration times and corrected peak areas. The non aqueous CE separation conditions and the spectral characteristics of 3,4-MDMA with respect to solvent and temperature effects are also discussed. PMID- 11499476 TI - Quantitation of entacapone glucuronide in rat plasma by on-line coupled restricted access media column and liquid chromatography-tandem mass spectrometry. AB - A column-switching liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS-MS) method was developed for the direct analysis of entacapone glucuronide in plasma. The plasma samples (5 microl) were injected onto a C18-alkyl-diol silica (ADS) column and the matrix compounds were washed to waste with a mixture of 20 mM ammonium acetate solution at pH 4.0-acetonitrile (97:3). The retained analyte fraction containing (E)- and (Z)-isomers of glucuronides of entacapone and tolcapone glucuronide (internal standard) was backflushed to the analytical C18 column, with a mixture of 20 mM ammonium acetate-acetonitrile (85:15) for the final separation at pH 7.0. The eluate was directed to the mass spectrometer after splitting (1:100). The mass spectrometer was operated in the negative ion mode and the deprotonated molecules [M-H]- were chosen as precursor ions for the analytes and internal standard. Collisionally induced dissociation of [M-H] in MS-MS resulted in loss of the neutral glucuronide moiety and in the appearance of intensive negatively charged aglycones [M-H-Glu]-, which were chosen as the product ions for single reaction monitoring. Quantitative studies showed a wide dynamic range (0.0025-100 microg/ml) with correlation coefficients better than 0.995. The method was repeatable within-day (relative standard deviation, RSD<7%) and between-day (RSD<14%) and the recovery (78-103%) was better than with the traditional, laborious pretreatment method. The use of tandem mass spectrometry permitted low limits of detection (1 ng/ml of entacapone glucuronide). The method was applied for the quantitation of (E)- and (Z)-isomers of entacapone glucuronide in plasma of rats used in absorption studies. PMID- 11499477 TI - High-performance affinity capture-removal of bacterial pyrogen from solutions. AB - Synthetic peptide S3delta has high affinity for bacterial endotoxin or lipopolysaccharide (LPS). Under tested conditions of pH 5-9 and 0-0.4 M NaCl, the affinity constant, KD ranged from 2.10(-6) to 2 x 10(-9) M(-1). A novel affinity matrix based on peptide S3delta was developed for removal of LPS from solutions such as: water; buffers with a wide range of ionic strength and pH; medium for cell culture; and protein solutions under optimized conditions. At a starting LPS of approximately 100 EU/ml, a post-purification level below 0.005 EU/ml was achieved. PMID- 11499478 TI - Quantitative determination of perifosine, a novel alkylphosphocholine anticancer agent, in human plasma by reversed-phase liquid chromatography-electrospray mass spectrometry. AB - A sensitive and selective reversed-phase LC-ESI-MS method to quantitate perifosine in human plasma was developed and validated. Sample preparation utilized simple acetonitrile precipitation without an evaporation step. With a Develosil UG-30 column (10 x 4 mm I.D.), perifosine and the internal standard hexadecylphosphocholine were baseline separated at retention times of 2.2 and 1.1 min, respectively. The mobile phase consisted of eluent A, 95% 9 mM ammonium formate (pH 8) in acetonitrile-eluent B, 95% acetonitrile in 9 mM ammonium formate (pH 8) (A-B, 40:60, v/v), and the flow-rate was 0.5 ml/min. The detection utilized selected ion monitoring in the positive-mode at m/z 462.4 and 408.4 for the protonated molecular ions of perifosine and the internal standard, respectively. The lower limit of quantitation of perifosine was 4 ng/ml in human plasma, and good linearity was observed in the 4-2,000 ng/ml range fitted by linear regression with 1/x weight. The total LC-MS run time was 5 min. The validated LC-MS assay was applied to measure perifosine plasma concentrations from patients enrolled on a phase I clinical trial for pharmacokinetic/pharmacodynamic analyses. PMID- 11499479 TI - Determination of an arylether antiarrhythmic and its N-dealkyl metabolite in rat plasma and hepatic microsomal incubates using liquid chromatography-tandem mass spectrometry. AB - A method was developed and validated for the quantification of (+/-)-trans-[2 morpholino-1-(1-naphthalene-ethyloxy]cyclohexane monohydrochloride (RSD1070) and its N-dealkyl metabolite in rat plasma and hepatic microsomal incubates. Chromatographic separations were achieved using reversed-phase high-performance liquid chromatography coupled with positive ion electrospray ionization and detection by tandem mass spectrometry. The assay was linear from 2.5 to 100 ng/ml and this range was used for validation. Inter- and intra-assay variability (n=6), extraction recovery, and stability in plasma were assessed. The estimated limit of quantitation was in the range 2.5-3 ng/ml for both analytes in rat plasma. The analytical method was used in a pharmacokinetic study of RSD1070 in rats after a single i.v. bolus of 12 mg/kg. PMID- 11499480 TI - Gas chromatography-combustion-isotope ratio mass spectrometry analysis of 19 norsteroids: application to the detection of a nandrolone metabolite in urine. AB - Determination of whether the major metabolite of nandrolone in urine, 19 norandrosterone (19-NA), is exogenous or endogenous in origin is one of the most exciting challenges for antidoping laboratories. Gas chromatography-combustion isotope ratio mass spectrometry (GC-C-IRMS) can be used to differentiate these two origins by carbon isotopic ratio analysis. A complete method for purification of 19-NA in urine has been established. Acetylated ketosteroids, and in particular 19-NA, are isolated from the urine matrix before analysis after hydrolysis and purification of urine by reversed-phase and normal solid-phase extraction. The limit of detection for 19-NA was about 60 ng with recoveries of 54-60%. Evidence of exogenous administration of 19-NA may be established from isotope ratio determination from the 13C/12C ratios of several synthetic 19 norsteroids compared to those obtained for endogenous steroids. PMID- 11499481 TI - Liquid chromatography-tandem mass spectrometric quantitation of cyclophosphamide and its hydroxy metabolite in plasma and tissue for determination of tissue distribution. AB - Cyclophosphamide (CP) and its metabolite, hydroxycyclophosphamide (OH-CP) have been quantitated in mouse plasma and tissue by derivatization combined with liquid chromatography-tandem mass spectrometry (LC-MS-MS). The derivatization was conducted immediately upon sample collection, to trap the OH-CP metabolite intermediate prior to further conversion to phosphoramide mustard or other reaction products. This simple and straightforward derivatization procedure, combined with sample extraction via protein precipitation, allowed quantitation of CP and the oxime derivative of OH-CP in plasma for concentrations ranging from approximately 12.5-3333 ng/ml, and in spleen tissue for concentrations of 1,250 50,000 ng/g. The short cycle time (2.5 min) of the LC-MS-MS method allowed high throughput analysis with minimal matrix interference. Mouse plasma levels were quantitated for doses of 40, 65 and 120 mg/kg; spleen concentrations were determined for mice dosed at 120 mg/kg. The CP and oxime plasma levels correlated well with dose amounts. The CP levels in the spleen and plasma were similar while the oxime levels in the spleen were significantly lower than the plasma. PMID- 11499482 TI - o-Phthalaldelhyde-N-acetylcysteine polyamine derivatives: formation and stability in solution and in C18 supports. AB - A comparative study of different derivatization procedures has been performed in order to improve the stability of the reaction products o-phthalaldehyde-N acetylcysteine (OPA-NAC) polyamines. Procedures such as solution derivatization, solution derivatization followed by retention on a packing support, derivatization on different packing supports and on-column derivatization, have been optimized and compared. The degradation rate constant (k) of the derivative was dependent on the procedure used and on the analyte. For the spermine (the most unstable isoindol tested) k was 8 +/- 2 x 10(-2) min(-1) in solution versus 7.7 +/- 1.1 x 10(-4) min(-1) on the (C18) solid support. The results obtained showed that forming the derivative on the packing support (C18) gave the best results following this procedure: conditioning the cartridges with borate buffer (1 ml, 0.5 M, pH 8), retention of the analyte, addition of 0.8 ml of OPA-NAC reagent, 0.2 ml borate buffer 0.8 M (pH 8) and elution of the isoindol with 3 ml of MeOH-borate buffer (9:1). The different derivatization procedures have been used to study the stability of the reaction products OPA-NAC polyamines formed in urine matrix using spermine as model compound. Similar results were obtained for standard solutions and urine samples. PMID- 11499483 TI - Rapid high-performance liquid chromatographic determination of docetaxel (Taxotere) in plasma using liquid-liquid extraction. AB - A new rapid and sensitive high-performance liquid chromatographic method for analysis of docetaxel (Taxotere) in human plasma was developed and validated. After adding an internal standard (paclitaxel, Taxol), plasma was extracted following a simple liquid-liquid extraction with diethyl ether. Extraction efficiency averaged 95% for docetaxel. Separation was performed using a Nucleosil (C18) 5 microm column, monitored at 227 nm. The isocratic mobile phase consisted of acetonitrile-acetate buffer, pH 5-tetrahydrofuran (45:50:5, v/v) pumped at a flow-rate of 1.8 ml/min. The limit of quantification for docetaxel in plasma was 12.5 ng/ml. Retention times for docetaxel and paclitaxel were 7.7 and 9 min, respectively. Standard curves were linear over a range of 25-1,000 ng/ml. This new method is rapid since it does not require time-consuming extraction procedures, or complex chromatographic conditions. This rapidity, along with the lack of chromatographic interferences with various other drugs likely to be administered to the cancer patients (pain killers, corticoids, antiemetics drugs) make this method suitable for daily routine analysis of Taxotere, a major anticancer drug extensively used in clinical oncology. PMID- 11499484 TI - Simultaneous determination of fluorinated inhalation anesthetics in blood by gas chromatography-mass spectrometry combined with a headspace autosampler. AB - Although the fluorinated inhalation anesthetics, including desflurane, sevoflurane, isoflurane, enflurane, and halothane are commonly used, fatal cases resulting from their abuse or misuse have been reported. To date, gas chromatography (GC) equipped with different kinds of detectors has been utilized to analyze inhalation anesthetics. However, none of them can detect desflurane reliably or analyze all five common anesthetics simultaneously. The purpose of the present work is to further modify the previously developed headspace (HS) GC MS method for blood isoflurane determination to analyze and distinguish five common clinical inhalation anesthetics, simultaneously. The modified HS-GC-MS method adopts a 60 m x 0.25 mm I.D., 0.25 microm film thickness DB-5 capillary column along with an adequate GC temperature program, which gives the five inhalation anesthetics, including isoflurane and its isomer, enflurane, a high resolution. The method also takes both the volatility and the influence of the top space on the obtained concentration into consideration and therefore keeps the sample loss acceptable even for analyzing the highly volatile desflurane. Within a certain concentration range of the calibration standard (about 20-300 microg/ml), this method shows a good linearity with correlation coefficients greater than 0.999. In addition, both within- and between-run precision and accuracy results meet the validation requirements as well as the tested results of practical blood samples of desflurane. In summary, this is a reliable analytical method to simultaneously determine the concentration of five common inhalation anesthetics in blood. Such a method is very practical for both clinical and occupational monitoring, as well as for analytical toxicology. PMID- 11499485 TI - A high-performance liquid chromatography assay with ultraviolet detection for olanzapine in human plasma and urine. AB - Olanzapine is a commonly used atypical antipsychotic medication for which therapeutic drug monitoring has been proposed as clinically useful. A sensitive method was developed for the determination of olanzapine concentrations in plasma and urine by high-performance liquid chromatography with low-wavelength ultraviolet absorption detection (214 nm). A single-step liquid-liquid extraction procedure using heptane-iso-amyl alcohol (97.5:2.5 v/v) was employed to recover olanzapine and the internal standard (a 2-ethylated olanzapine derivative) from the biological matrices which were adjusted to pH 10 with 1 M carbonate buffer. Detector response was linear from 1-5000 ng (r2>0.98). The limit of detection of the assay (signal:noise=3:1) and the lower limit of quantitation were 0.75 ng and 1 ng/ml of olanzapine, respectively. Interday variation for olanzapine 50 ng/ml in plasma and urine was 5.2% and 7.1% (n=5), respectively, and 9.5 and 12.3% at 1 ng/ml (n=5). Intraday variation for olanzapine 50 ng/ml in plasma and urine was 8.1% and 9.6% (n=15), respectively, and 14.2 and 17.1% at 1 ng/ml (n=15). The recoveries of olanzapine (50 ng/ml) and the internal standard were 83 +/- 6 and 92 +/- 6% in plasma, respectively, and 79 +/- 7 and 89 +/- 7% in urine, respectively. Accuracy was 96% and 93% at 50 and 1 ng/ml, respectively. The applicability of the assay was demonstrated by determining plasma concentrations of olanzapine in a healthy male volunteer for 48 h following a single oral dose of 5 mg olanzapine. This method is suitable for studying olanzapine disposition in single or multiple-dose pharmacokinetic studies. PMID- 11499486 TI - Validation of a high-performance liquid chromatography method for tramadol and o desmethyltramadol in human plasma using solid-phase extraction. AB - An HPLC system using solid-phase extraction and HPLC with UV detection has been validated in order to determine tramadol and o-desmethyltramadol (M1) concentrations in human plasma. The method developed was selective and linear for concentrations ranging from 50 to 3,500 ng/ml (tramadol) and 50 to 500 ng/ml (M1) with mean recoveries of 94.36 +/- 12.53% and 93.52 +/- 7.88%, respectively. Limit of quantitation (LOQ) was 50 ng/ml. For tramadol, the intra-day accuracy ranged from 95.48 to 114.64% and the inter-day accuracy, 97.21 to 103.24%. Good precision (0.51 and 18.32% for intra- and inter-day, respectively) was obtained at LOQ. The system has been applied to determine tramadol concentrations in human plasma samples for a pharmacokinetic study. PMID- 11499487 TI - Immunoassay for human serum albumin using capillary electrophoresis-semiconductor laser-induced fluorometry. AB - Capillary electrophoresis combined with semiconductor laser-induced fluorometry was applied to an immunoassay of human serum albumin. Human serum albumin was labeled with a fluorescent molecule (Cy5), which has an absorption maximum at 649 nm. The labeled albumin was purified by ultrafiltration in order to reduce signals, which are unreacted labeling reagent, product, and fragment products derived there from. After the purification, no signal for unreacted labeling reagent and fragment products was detectable in the electropherogram of the labeled albumin. The labeled albumin was then reacted with anti-albumin to form an immunocomplex, which was separated from the excess free albumin. The competitive immunoassay was used in the determination of human serum albumin in a controlled serum sample, using the labeled albumin. The obtained value was found to be 0.21 +/- 0.02 mg/ml, which is in good agreement with other known values. PMID- 11499488 TI - Simple high-performance liquid chromatographic method to analyze serum creatinine has several advantages over the Jaffe picric acid reaction as demonstrated with a cimetidine dose response in rhesus monkeys. AB - A simple method for creatinine determination was developed using high-performance liquid chromatography (HPLC) to more accurately monitor serum creatinine levels in experimental animal models when compared to the Jaffe method. The new HPLC procedure will replace the traditional Jaffe method for rhesus monkey kidney function studies. We developed an isocratic method using a polymeric, hydrophilic, silica-based strong cation-exchange bed with a 5.0 mmol/l lithium acetate matrix, pH 4.9, which isolates creatinine with no detectable impurities as determined by three-dimensional ultraviolet-visible spectral analysis. Sample preparation includes deproteination with acetonitrile, evaporation, and resolubilization in mobile phase followed by quantitation with UV detection at 234 nm. Extraction efficiency across the measured range was 96 +/- 2%. From numerous extracted rhesus monkey creatinine curves (n=38) a slope of 251,100 +/- 756 (95% CI) and an intercept of 675.6 +/- 712.7 (95% CI) was calculated. Extraction efficiency and peak purity tests with human plasma were cross-compared with rhesus monkey serum producing equivalent results. An average of 120 samples can be run daily. PMID- 11499489 TI - Analysis of urinary N-acetyl-S-(N-methylcarbamoyl)cysteine, the mercapturic acid derived from N,N-dimethylformamide. AB - Human biotransformation of the industrial solvent N,N-dimethylformamide gives raise to N-acetyl-S-(N-methylcarbamoyl)cysteine (AMCC) which has the longest half life (about 23 h) among urinary metabolites of N,N-dimethylformamide. It could be used for monitoring industrial exposure over several workdays, by measuring it in urine samples collected at the end of the working week. This is consistent with the suggestions of the American Conference of Governmental Industrial Hygienists, which established a limit of 40 mg/l for the year 2000. An easy, cheap and user friendly method has been developed for determination of urinary AMCC. Unlike currently available methods, it requires neither a time-consuming preparation phase nor gas chromatographic analysis with a nitrogen-phosphorus or mass detector. The method uses high-performance liquid chromatography (HPLC), with an UV detector at 436 nm. A 10-microl volume of urine is added to a carbonate hydrogen carbonate buffer and mixed with a dabsyl chloride solution in acetonitrile. The reaction between AMCC and the reagent is performed at 70 degrees C for 10 min. The 'dabsylated' product is stable for at least 12 h. After brief centrifugation, the solution is ready for HPLC analysis using a C18 column (250 x 4.6 mm, 5 microm). The method is sensitive (detection limit 1.8 mg/l) and specific. It identified urinary AMCC in urine of 40 subjects not exposed to N,N dimethylformamide with a median concentration of 3.9 mg/l. In urine samples from 20 workers exposed to N,N-dimethylformamide (5-40.8 mg/m3), AMCC concentrations ranged from 16 to 170 mg/l. Industrial toxicology laboratories with limited instrumentation will be able to use it in the biological monitoring of workers exposed to N,N-dimethylformamide. PMID- 11499490 TI - Solid-phase analysis method for (S)-[18F]fluorocarazolol and its metabolites. AB - (S)-[18F]Fluorocarazolol is a radiopharmaceutical developed to quantitatively assess beta-adrenergic receptors in vivo via positron emission tomography imaging. Since radioactive metabolites of (S)-[18F]fluorocarazolol rapidly appear in the plasma, methods for conveniently and reliably evaluating plasma for (S) [18F]fluorocarazolol content are required. Here we present methods and validation of an approach using commercial extraction cartridges that is faster and more convenient than an approach using internal-surface reverse-phase chromatography but yields comparable results. PMID- 11499491 TI - Determination of plasma bromvalerylurea and its main metabolite by a simple high performance liquid chromatographic method and quantitation of bromide by energy dispersive X-ray spectrometry in carbon tetrachloride-intoxicated rats. AB - In the present study, small volumes of plasma were used for the measurement of bromvalerylurea (BVU), its metabolite, 3-methylbutyrylurea (MVU), and bromide in carbon tetrachloride (CCl4)-treated rats by HPLC-UV and energy dispersive X-ray spectrometry. A liquid-liquid extraction system was also investigated. BVU and MVU were extracted from 100 microl plasma samples in a single-step involving deproteination with 1 M hydrochloric acid using ethenzamide as internal standard. Samples were separated by HPLC in an acetonitrile-8 mM potassium dihydrogenphosphate buffer (35:65, v/v) mobile phase at a flow-rate of 0.4 ml/min on a 15 cm octadecylsilyl column at room temperature. Analytes were detected at a wavelength of 210 nm. The limits of quantitation for BVU, MVU and bromide are 0.1, 0.1 and 50 microg/ml, respectively. The intra-day accuracies over the range of concentrations were 95.8 to 121.1%, 97.2 to 119.7% and 96.2 to 105.8% for BVU, MVU and bromide, respectively. The inter-day accuracies were 97.7 to 115.1%, 98.3 to 111.6% and 98.3 to 102.9% for BVU, MVU and bromide, respectively. The absolute recoveries using tert.-butyl methyl ether are 96-98% for BVU and 95-98% for MVU. The decline in the plasma concentrations of BVU in olive oil-treated rats fitted a one-compartment model and the plasma MVU level reached a peak at around 1.5-2 h and then decreased gradually. The elimination of BVU in CCl4 (1 ml/kg)-treated rats was delayed and MVU production was less than that in the olive oil-treated group. However, there was no difference in the plasma levels of bromide between CCl4-treated rats and control rats. rights reserved. PMID- 11499492 TI - Distributive justice and clinical trials in the Third World. AB - One of the arguments against conducting human subject trials in the Third World adopts a distributive justice principle found in a commentary of the CIOM'S Eighth Guideline for international research on human subjects. Critics argue that non-participant members of the community in which the trials are conducted are exploited because sponsoring agencies do not ensure that the products developed have been made reasonably available to these individuals. I argue that the distributive principle's wording is too vague and ambiguous to be used to criticize any trial. Furthermore, the mere fact that an experiment does not fulfill this particular distributive justice principle does not entail that it is unethical. PMID- 11499493 TI - Distributive justice and clinical trials in the Third World. PMID- 11499494 TI - Defining and delineating a duty to prognosticate. AB - Prognostication, the process of formulating and communicating a prognosis, is no longer considered by most physicians to be an essential task in caring for patients with serious illness. Because of this fact, it is not surprising to find that when physicians attempt to engage in prognostication, they do it poorly. What may be surprising to those outside the medical community is the extent to which professional norms have developed which actively discourage physicians from engaging in prognostication. This article explores the causes of this state of affairs and the justifications offered for it. The conclusion is reached that physicians have a professional responsibility to competently engage in prognostication based upon the doctrine of informed consent, and that a failure or refusal to do so has not only potential legal ramifications, but serious negative implications for many of the core issues in bioethics, such as the use of advance directives, palliative medicine, and medical futility. PMID- 11499496 TI - Complexity of the concept of disease as shown through rival theoretical frameworks. AB - The concept of disease has been the subject of a vast, vivid and versatile debate. Categories such as "realist", "nominalist", "ontologist", "physiologist", "normativist" and "descriptivist" have been applied to classify disease concepts. These categories refer to underlying theoretical frameworks of the debate. The objective of this review is to analyse these frameworks. It is argued that the categories applied in the debate refer to profound philosophical issues, and that the complexity of the debate reflects the complexity of the concept itself: disease is a complex concept, and does not easily lend itself to definition. PMID- 11499495 TI - Contending medical decision models. PMID- 11499497 TI - A compassionate autonomy alternative to speciesism. AB - Many people in the animal welfare community have argued that the use of nonhuman animals in medical research is necessarily based on speciesism, an unjustified prejudice based on species membership. As such it is morally akin to racism and sexism. This is misguided. The combined capacities for autonomy and sentience with the obligations derived from relations support a morally justifiable rationale for using some nonhuman animals in order to limit the risk of harm to humans. There may be a few cases where it is morally better to use a never sentient human than a sentient animal, but these cases are few and would not fulfill the current need for research subjects. The use of nonautonomous animals instead of humans in risky research can be based on solid moral ground. It is not necessarily speciesism. PMID- 11499498 TI - A moral economy of American medicine in the managed-care era. AB - The moral economy of American medicine has been transformed by contentious innovations in organization, administration, regulation, and finance. In many settings old fee-for-service incentives and disincentives have been replaced by those of "managed care", while in other settings they have been diluted or distorted. In the everyday care of patients, old and new may alternate or interact. These innovations may also be having secondary effects on participation in life-sciences research and the development and employment of new technologies, discouraging collective support for preliminary investigation and delaying adoption of improved goods and services until cost-reducing potential has already been realized. This motivational complexity, particularly in its moral dimensions, is hard to address using standard assumptions and methods. I argue for different assumptions, based on the clinical behavior of individual patients rather than the market behavior of aggregated consumers, and I describe a different method, based on an old idea in political economy. I then present a new way to explain the core obligations of clinicians, researchers, and planners and to interpret the policy problems they must now share. PMID- 11499499 TI - Pantagruelism: a Rabelaisian inspiration for understanding poisoning, euthanasia and abortion in the Hippocratic oath and in contemporary clinical practice. AB - Contrary to the common view, this paper suggests that the Hippocratic oath does not directly refer to the controversial subjects of euthanasia and abortion. We interpret the oath in the context of establishing trust in medicine through departure from Pantagruelism. Pantagruelism is coined after Rabelais' classic novel Gargantua and Pantagruel. His satire about a wonder herb, Pantagruelion, is actually a sophisticated model of anti-medicine in which absence of independent moral values and of properly conducted research fashion a flagrant over medicalization of human problems. Ultimately this undermines the therapeutic core of medicine itself. We contend that PAS is a case of such over-medicalization and that its institution creates medicophobia. This article does not express an opinion about euthanasia in general. Rather, we claim that physicians should learn from the oath and from Rabelais that they should keep their practice to medical care and not to exploit their expertise and social privileges for the sake of ulterior motives, even when their patients desire those goals. PMID- 11499500 TI - The unwillingness of some researchers to share bacterial strains and reagents. PMID- 11499501 TI - Estrogenicity of fissure sealants and adhesive resins determined by reporter gene assay. PMID- 11499502 TI - Glucosyltransferase inactivation reduces dental caries. PMID- 11499503 TI - Collaboratories: leveraging information technology for cooperative research. AB - Economic, organizational, and societal pressures, as well as the desire to reach shared goals more efficiently and effectively, are driving an increase in collaborative research. Research collaborations frequently occur among participants separated by temporal, geographical, organizational, disciplinary, and cultural boundaries. Increasingly complex collaborative projects focus attention on the question of how to facilitate working together. Through so called collaboratories, information technology can play an important role in addressing this question. A collaboratory can be defined as an information technology infrastructure that supports cooperation among individuals, groups, or organizations in pursuit of a shared goal by facilitating interaction, communication, and knowledge-sharing. Tools such as Web-based collaborative workspaces, Internet discussion lists/newsgroups/real-time chat, screen- and application-sharing, Web-based conferencing, online Web page mark-up, automatic notification, and videoconferencing can be used to implement collaboratories. Collaboratories have significant potential to facilitate cooperative research, but should be evaluated carefully to determine best practices. PMID- 11499504 TI - Equivalence testing with dental clinical trials. AB - Clinical trials whose purpose are to show equivalence of two or more treatments have traditionally utilized methods for demonstrating superiority, and, if no statistical differences are found, the treatments are assumed to be equivalent. Methods for directly demonstrating equivalence have been developed by the pharmaceutical industry to establish the bioequivalence between drug formulations. This approach should have significant future implications for dental research. It is presently well-characterized in the statistical literature, but is virtually unknown within the dental literature and has had limited documented application with dental clinical trials. The purpose of this paper is to consider equivalence testing with dental clinical trials. Equivalence and superiority testing are contrasted, and comparisons between parallel and crossover designs with equivalence testing are discussed. To exemplify equivalency trial methodology for dental research application, several examples are proposed, including, in detail, an implant overdenture clinical trial to test the equivalence of treatment outcome between a two- and a four-implant overdenture treatment. Sample size calculations for otherwise-identical crossover and parallel equivalence studies are demonstrated. Results show that for a power of 0.90, a sample size of 24 is required for the crossover trial and over 150 subjects for the parallel design. For this dental research application, the equivalence/crossover design methodology is shown to be an efficient and appropriate approach. PMID- 11499505 TI - Vanilloid receptor expression and capsaicin excitation of rat dental primary afferent neurons. AB - Little is known about the molecular mechanisms that cause excitation of neurons which innervate the teeth. We investigated whether rat dental sensory neurons express the vanilloid (capsaicin) receptor (VR1). Dental sensory neurons were identified by retrograde transport of the fluorescent dye DiIC18 placed in maxillary molars. Patch-clamp recordings in culture showed that 65% of DiIC18 labeled rat trigeminal ganglion neurons are excited by capsaicin. Responders covered the entire range of cell sizes examined (soma diameter, 24 to 48 microm). All non-responders had a soma diameter > 33 microm. Capsazepine (1 microM) reduced the capsaicin-evoked membrane current (6/6) and depolarization (7/7 responders). RT-PCR amplified a 375-bp product from DiIC18-labeled neurons which was identical to that expected for VR1. Thus, many rat dental primary afferent neurons are excited by capsaicin, and the response appears to be mediated by VR1. These results suggest that pharmacological blockers of VR1 may provide significant relief of dental pain. PMID- 11499506 TI - Sustained microglial immunoreactivity in the caudal spinal trigeminal nucleus after formalin injection. AB - Recent studies indicate that glia may be involved in altered nociceptive processing after a peripheral inflammatory lesion produced by injection of inflammatory reagents such as formalin and zymosan. Most of these studies, however, confined their observations to a period shortly after the injections. This study investigated the immunohistochemical responses of microglia in the caudal part of the spinal trigeminal nucleus for up to 60 days after subcutaneous injection of formalin into the lateral faces of Wistar rats. The results showed obvious up-regulation of microglial markers such as OX-18, OX-42 and OX-6 up to 21 days after formalin injection. These were somewhat reduced at 30 days after injection. Electron microscope investigation revealed no evidence of significant phagocytosis of degenerative neuronal elements by microglia in the nucleus at the time--that is, 7 days after formalin injection, when microglial activation was at its peak. Significantly, however, the period of microglial activation corresponded closely to that showing enhanced nociceptive behavior after perioral formalin injection (Cadet et al., 1995). This study indicates a microglial role in the genesis of enhanced nociceptive behavior. PMID- 11499507 TI - Primary and secondary induction of apoptosis in odontoblasts after cavity preparation of rat molars. AB - The death regulation of damaged pulp cells after cavity preparation is not well known. In this study, we examined whether apoptosis is associated with the death regulation of damaged pulp cells. In normal rat molars, terminal deoxynucleotidyl transferase-mediated labeling (TUNEL)-positive cells were not observed. Just after surgery, odontoblasts under cavities were TUNEL-positive, and these signals disappeared in six hours. One day after surgery, we found the reappearance of TUNEL-positive cells in the subodontoblastic region under cavities, and positive signals disappeared in four days. Ultrastructure of TUNEL-positive cells showed characteristics typical of apoptotic cells. Phagocytosis of apoptotic cells by scavenger cells was also observed. By immunohistochemistry, we also found Bcl-2 positive odontoblasts one day after surgery. These results suggest that two waves of apoptosis are induced in odontoblasts after cavity preparation, and that apoptotic cells must be eliminated before the initiation of reparative dentinogenesis. PMID- 11499508 TI - Tumor necrosis factor-alpha and its receptors, p55 and p75, in gingiva of adult periodontitis. AB - Tumor necrosis factor-alpha (TNF-alpha), a pro-inflammatory cytokine, can stimulate matrix metalloproteinase synthesis and osteoclastic bone resorption. We hypothesized that elevated expression of TNF-alpha and its p55 and p75 receptors (TNF-R) in gingival tissue might associate with periodontitis. Immunohistochemistry was used for the study of the localization of TNF-alpha and its p55 and p75 TNF-R in adult periodontitis (AP) gingival tissue, in comparison with that in healthy control specimens. TNF-alpha and p55 TNF-R were detected in sulcular epithelial basal cells and in monocyte/macrophages, fibroblasts, and endothelial cells in the AP gingival tissue specimens, but mainly in fibroblasts and endothelial cells in control specimens. P75 TNF-R was occasionally found in monocyte/macrophage-like cells in gingival tissue specimens. The percentage of TNF-alpha-containing cells was not increased in AP compared with controls (13.2%+/-6.1% vs. 12.8%+/-7.6%), but, due to the increased cellularity of AP samples, the number of TNF-alpha positive cells/mm2 was clearly increased (1621+/ 663 vs. 664+/-191, p > 0.001). Thus, AP gingival tissue has an elevated expression of TNF-alpha and especially its p55 receptor, suggesting that TNF alpha may contribute to tissue degradation in periodontitis. PMID- 11499509 TI - Osteoblast integrin adhesion and signaling regulate mineralization. AB - Integrin adhesion and signaling events may contribute to the progressive differentiation of the osteoblast and to the initiation of a mineralized matrix. The purpose of our study was to begin to analyze the role of integrin receptors, in particular alpha2beta1, alpha5beta1, and alphaVbeta3, regarding mediation of the initiation of a mineralized matrix. Integrin-perturbation assays were conducted in microdot cultures of UMR-106-01 Bone Sialoprotein (BSP) osteoblast cells. For phenotypic analysis, we performed bright-field microscopy and Aliziran Red S staining to analyze effects on mineralization initiation. Mineralization was reduced significantly (P < 0.001) following the addition of alpha5- or beta1 integrin subunit antibody by approximately 20% and 45%, respectively--alphaVbeta3 integrin by nearly 65%, and alpha2beta1 integrin by nearly 95%. This effect was reversible following the removal of the antiintegrin antibody. These results suggest that integrin adhesion and signaling events may contribute to the ability of this cell line to mediate the initiation of the mineralization phenotype biologically. PMID- 11499510 TI - The effects of MMP inhibitors on human salivary MMP activity and caries progression in rats. AB - Previous studies suggest that salivary and pulp-derived host enzymes, matrix metalloproteinases (MMPs), may be involved in dentin caries pathogenesis. To study the inhibition of acid-activated human salivary MMPs by non-antimicrobial chemically modified tetracyclines (CMTs), we used a functional activity assay with 125I-labeled gelatin as a substrate. To address the role of MMPs in the progression of fissure caries in vivo, we administered the MMP inhibitors CMT-3 and zoledronate to young rats per os for 7 weeks, 5 days a week. Caries lesions were visualized by Schiff reagent in sagittally sectioned mandibular molars. Marked reduction in gelatinolytic activity of human salivary MMPs was observed with CMT-3. CMT-3 and zoledronate, both alone and in combination, also reduced dentin caries progression in the rats. These results suggest that MMPs have an important role in dentin caries pathogenesis, and that MMP inhibitors may prove to be useful in the prevention of caries progression. PMID- 11499511 TI - Salivary non-immunoglobulin agglutinin inhibits human leukocyte elastase digestion of acidic proline-rich salivary proteins. AB - Saliva contains acidic proline-rich salivary proteins that are involved in the formation of the salivary pellicle coating supragingival tooth surfaces. However, human leukocyte elastase, arriving in gingival exudates from inflamed periodontal tissues, degrades the acidic proline-rich salivary proteins, preventing binding to hydroxylapatite surfaces. Here it is reported that high-molecular-weight non immunoglobulin salivary agglutinin inhibited the proteolytic action of human leukocyte elastase on purified acidic proline-rich salivary proteins. Inhibition was eliminated with monoclonal antibody to a protein determinant on the salivary agglutinin. The addition of antibody against salivary agglutinin blocked the inhibitory effect of parotid saliva on exogenously applied human leukocyte elastase, allowing for the elastase-mediated digestion of the salivary acidic proline-rich salivary proteins. Salivary agglutinin, therefore, is a physiologically important inhibitor of human leukocyte elastase and is able to inhibit elastase-mediated digestion of salivary acidic proline-rich proteins. PMID- 11499512 TI - Intrafibrillar mineral may be absent in dentinogenesis imperfecta type II (DI II). AB - High-resolution synchrotron radiation computed tomography (SRCT) and small-angle x-ray scattering (SAXS) were performed on normal and dentinogenesis imperfecta type II (DI-II) teeth. The SRCT showed that the mineral concentration was 33% lower on average in the DI-II dentin with respect to normal dentin. The SAXS spectra from normal dentin exhibited low-angle diffraction peaks at harmonics of 67.6 nm, consistent with nucleation and growth of the apatite phase within gaps in the collagen fibrils (intrafibrillar mineralization). In contrast, the low angle peaks were almost non-existent in the DI-II dentin. Crystallite thickness was independent of location in both DI-II and normal dentin, although the crystallites were significantly thicker in DI-II dentin (6.8 nm [SD = 0.5] vs. 5.1 nm [SD = 0.6]). The shape factor of the crystallites, as determined by SAXS, showed a continuous progression in normal dentin from roughly one-dimensional (needle-like) near the pulp to two-dimensional (plate-like) near the dentin enamel junction. The crystallites in DI-II dentin, on the other hand, remained needle-like throughout. The above observations are consistent with an absence of intrafibrillar mineral in DI-II dentin. PMID- 11499513 TI - High-resolution electron microscopy of the crystallites of fossil enamels obtained from various geological ages. AB - To elucidate the stability of the central dark line (CDL) in biologically induced hydroxyapatite crystals, we examined the diagenetic changes on the microstructures of the crystallites during the course of fossilization. Using transmission electron microscopy, we investigated the enamel crystallites of fossil animals of various geological ages ranging from Pleistocene to Cretaceous. Electron micrographs indicated that the microstructures and lattice images of each crystallite in fossil enamels were well-preserved regardless of the thickness of the enamel layer, and the presence of CDLs in fossil enamel crystals was also confirmed. The results indicated that the microstructure of hydroxyapatite crystals containing lattice images of CDLs appear stable during long geological periods. In addition, we conclude that the existence of lattice images in apatite with CDLs may be an indicator for the assessment of the evolution of dental enamel from fossil remains. PMID- 11499514 TI - Adhesion to and decalcification of hydroxyapatite by carboxylic acids. AB - Fundamental to the processes of decalcification of or adhesion to mineralized tissues is the molecular interaction of acids with hydroxyapatite. This study was undertaken to chemically analyze the interaction of 1 mono-, 2 di-, 1 tri-, and 2 polycarboxylic acids with hydroxyapatite in an attempt to elucidate the underlying mechanism. Maleic, citric, and lactic acid decalcified hydroxyapatite, in contrast to oxalic acid and the two polycarboxylic acids that were chemically bonded to hydroxyapatite. Solubility tests showed that the calcium salts of the former were very soluble, whereas those of the latter could hardly be dissolved in the respective acid solutions. Based on these data, an adhesion/decalcification concept was advanced that predicts that carboxylic acids, regardless of concentration/pH, either adhere to or decalcify hydroxyapatite, depending on the dissolution rate of the respective calcium salts in the acid solution. This contrasting behavior of organic acids most likely results from their differential structural conformations. PMID- 11499515 TI - Influence of pulse-delay curing on softening of polymer structures. AB - Resin composites may be polymerized according to one of several light-curing modes. These modes include variations in intensity of the curing lights and time delay before final cure. The so-called pulse-delay method has earlier been found to reduce the formation of gaps due to polymerization contraction, without mechanical properties of the resin composite being compromised. It was hypothesized that the slow pre-cure of this method would give rise to a different polymer structure than results when the polymer is cured in one step at high intensity. It was found that although the quantity of remaining double bonds was unaffected, the pulse-delay technique led to polymers of increased susceptibility to softening in ethanol. The softness increased with the intensity of the pre cure and with the waiting time before final cure. The increased softening may be interpreted as the manifestation of a polymer structure having fewer crosslinks. PMID- 11499516 TI - Evidence of a critical leucite particle size for microcracking in dental porcelains. AB - The leucite particles in dental porcelains are often partially encircled by microcracks that are the result of the thermal expansion mismatch between leucite and the surrounding glass matrix. Although the magnitude of the stress at the particle-matrix interface is independent of the particle size (Selsing, 1961), Davidge and Green (1968) showed experimentally that there is a critical particle size below which microcracking is absent. The critical particle size is explained by a Griffith-type energy balance criterion: Below the critical size, the stress magnitude may be sufficient to cause cracking, but there is insufficient strain energy for the creation of the new surfaces of the microcrack. The purpose of the present study was to determine whether the mean leucite particle size of a dental porcelain influences the degree of microcracking in the porcelain. Microcrack density, leucite particle surface area per unit volume, and leucite mean volume surface diameter, D3,2, were determined by quantitative stereology on 10 specimens each of 6 dental porcelains and Component No. 1 of the Weinstein et al. patent (US Patent 3,052,982, 1962). The fraction of leucite particles with microcracks around them, f(mc), was estimated for each porcelain from the microcrack density and the leucite surface area. Using the equations of Selsing (1961) and Davidge and Green (1968), we calculated the critical particle diameter, Dc, for leucite to be 4 microm. The porcelains were partitioned according to whether their mean leucite particle diameters, D3,2, fell above or below Dc, and their values of f(mc) were analyzed by a permutation test with random re-sampling. The porcelains with mean leucite particle diameters below Dc had a significantly lower fraction of cracked particles compared with the porcelains with mean leucite particle diameters above Dc (p < 0.05). This study provides evidence that microcracking in dental porcelain can be minimized by a reduction of the mean leucite particle diameter to less than 4 microm. PMID- 11499517 TI - Early-onset periodontitis in Morocco is associated with the highly leukotoxic clone of Actinobacillus actinomycetemcomitans. AB - A particular clone (JP2) of Actinobacillus actinomycetemcomitans with increased leukotoxin production has been isolated from individuals with early-onset periodontitis (EOP). The aim of this study was to determine the frequency of carriers of this clone and its association with EOP in Moroccan schoolchildren. Of 217 plaque samples, 131 (60.4%) were culture-positive for A. actinomycetemcomitans. A total of 19 of these isolates had a 530-bp deletion in the leukotoxin promoter region characteristic of the JP2 clone. A strong association between the presence of A. actinomycetemcomitans with the 530-bp deletion and EOP was found (adjusted OR = 29.4; 95% Cl = 8.3 - 104.4; p < 0.0005), while no association could be demonstrated between the presence of A. actinomycetemcomitans without the deletion and EOP (adjusted OR = 1.3; 95% CI = 0.5 -2.9; p = 0.750). The study demonstrates that the endemic presence, in a human population, of the highly leukotoxic JP2 clone may result in an unusually high prevalence of EOP. PMID- 11499518 TI - The effects of duration and intensity of stimulation on total protein and mucin concentrations in resting and stimulated whole saliva. AB - The present investigation has characterized the influence of the duration and intensity of stimulation on the secretion pattern of total protein and salivary mucins MG1 and MG2 in whole saliva. Resting and stimulated whole saliva was collected from six healthy subjects on 2 consecutive days. Whole saliva was collected for 2 five-minute intervals under resting conditions followed by collection under masticatory stimulation induced by the chewing of parafilm (1 g) at 10 or 60 strokes/min for 15 min. Flow rates were different under the 2 levels of stimulation. The concentration of total protein was different in resting and stimulated whole saliva but was not affected by the duration or intensity of stimulation. Analysis of mucin concentrations determined by capture ELISAs revealed that the pattern of MG1 secretion was similar to that of total protein. The pattern of MG2 secretion was unique in that no differences were observed in the concentration of this mucin under resting and stimulated conditions. This study shows that the pattern of protein secretion in whole saliva does not reflect the combined pattern observed for protein secretion in parotid and submandibular/sublingual glands, and that the secretion patterns of MG1 and MG2 in whole saliva are quite different from one another. PMID- 11499519 TI - Psychological disorder, conditioning experiences, and the onset of dental anxiety in early adulthood. AB - Most studies examining the origins of dental fear and anxiety have relied on cross-sectional data. These are subject to several problems, such as recall and uncertainty concerning temporal relationships. This paper uses longitudinal data from the Dunedin Multidisciplinary Health and Development Study to assess risk factors for the development of dental anxiety in persons between the ages of 18 and 26 years. It was hypothesized that psychological factors would be as important as conditioning experiences in the genesis of dental anxiety over this period. The eight-year incidence of dental anxiety was 16.5%. Five variables entered models predicting onset: multiple fears, symptoms of substance dependence, previous experience of invasive dental treatment, dental visiting pattern, and the extraction of one or more teeth. Separate analyses for those avoiding and those using dental services resulted in different explanatory models. These results indicated that both psychological and conditioning variables contributed to the development of dental anxiety in this population of young adults. PMID- 11499520 TI - "Come on in". PMID- 11499521 TI - Detection of nerve action potentials under low signal-to-noise ratio condition. AB - We propose a method for detection of action poten. tials (APs) under low signal to-noise ratio condition. It is based on multiresolution analysis. Three parameters are used for detection. Two of them are for determining if there is an AP or not, and the other is for the estimation of waveforms. Our method provides better estimated waveforms than the conventional de-noising approach. PMID- 11499522 TI - Heart instantaneous frequency (HIF): an alternative approach to extract heart rate variability. AB - Our study focuses on a new method of estimating the heart rate variability (HRV) which does not require the use of electrocardiogram (ECG) R-wave detection. Contrary to the R-wave detection method which requires a sampling frequency higher than 100 Hz, the one proposed here can be used to calculate the HRV from an ECG signal sampled at a frequency of approximately 5 Hz with a relative mean error of 0.03. This new method is based on extracting the instantaneous fundamental frequency from the ECG. The method could be efficiently used to extract the HRV from an ECG measured for healthy subjects performing an exercise in which the HRV increases linearly with time, and for subjects with respiratory and cardiac problems. The overall error decreased as we low-pass filtered the HRV with lower cut-off frequencies. Moreover, it was shown that the method could be efficiently used to calculate the HRV from blood pressure measurements and to be robust to noise. PMID- 11499523 TI - Bionic wavelet transform: a new time-frequency method based on an auditory model. AB - In this paper, a new adaptive wavelet transform, named bionic wavelet transform (BWT), is developed based on a model of the active auditory system. The most distinguishing characteristic of BWT is that its resolution in the time-frequency domain can be adaptively adjusted not only by the signal frequency but also by the signal instantaneous amplitude and its first-order differential. The automatically adjusted resolution, even in a fixed frequency along the time-axis, is achieved by introducing the active control of the auditory system into the wavelet transform (WT). Other properties of BWT include that: 1) BWT is a nonlinear transform that has high sensitivity and frequency selectivity; 2) BWT represents the signal with a concentrated energy distribution; and 3) the inverse BWT can reconstruct the original signal from its time-frequency representation. In order to compare these three properties between BWT and WT, experiments were conducted on both constructed signals and real speech signals. The results show that BWT performs better than WT in these three aspects, and that BWT is appropriate for speech signal processing, especially for cochlear implants. PMID- 11499524 TI - Analytical solution for pulsatile axial flow velocity waveforms in curved elastic tubes. AB - An analytical solution for pulsatile axial flow velocity waveforms in curved elastic tubes is presented. The result is obtained by exact solution of linearized Navier-Stokes and tube motion equations in a torroidal coordinate system. Fourier analysis is used to divide the flow into constant and oscillatory components which are separately considered. The solution is used to investigate the effects of curvature on volumetric axial velocity flow waveforms, as would be measured by Doppler ultrasound techniques. In typical human arteries, the greatest effects of curvature on the volumetric axial flow are exerted on the constant component and at low values of the frequency parameter for the oscillatory components. Here, the magnitude and phase angle of oscillatory flow in the curved tube, relative to that in the straight tube, differ by maximum values of 1.2% and 0.15 rad, respectively. However, constant flow may vary by as much as 60% at high Dean numbers. The solution is presented in a form similar to Womersley's solution for the straight elastic tube and may, thus, be incorporated into a transmission-line analog model. These models are frequently used to investigate axial flow velocity variations in mamillian circulatory systems and this work offers a tool which may extend these models to incorporate the effects of curvature. PMID- 11499525 TI - Modeling and closed-loop control of hypnosis by means of bispectral index (BIS) with isoflurane. AB - A model-based closed-loop control system is presented to regulate hypnosis with the volatile anesthetic isoflurane. Hypnosis is assessed by means of the bispectral index (BIS), a processed parameter derived from the electroencephalogram. Isoflurane is administered through a closed-circuit respiratory system. The model for control was identified on a population of 20 healthy volunteers. It consists of three parts: a model for the respiratory system, a pharmacokinetic model and a pharmacodynamic model to predict BIS at the effect compartment. A cascaded internal model controller is employed. The master controller compares the actual BIS and the reference value set by the anesthesiologist and provides expired isoflurane concentration references to the slave controller. The slave controller maneuvers the fresh gas anesthetic concentration entering the respiratory system. The controller is designed to adapt to different respiratory conditions. Anti-windup measures protect against performance degradation in the event of saturation of the input signal. Fault detection schemes in the controller cope with BIS and expired concentration measurement artifacts. The results of clinical studies on humans are presented. PMID- 11499526 TI - The biocompatibility, integrity, and positional stability of an injectable microstimulator for reanimation of the paralyzed larynx. AB - The biocompatibility, integrity, positional stability, and potential use of hermetically sealed injectable wireless micromachined microstimulators were investigated for reanimation of the paralyzed larynx. The device, consisting of silicon and glass, has been tested and proven to be biocompatible with no evidence of pathological tissue reaction or rejection up to one-year implantation in the rat dorsum and canine larynx. By one month, each unit was encapsulated by a thin membrane, which thickened to form a fibrous layer of less than 500 microm at 6-12 months. The microstimulators demonstrated long-term in vivo durability: the hermetic seal of only one in ten devices was breached. Once implanted, migration of the device varied from 0 to 3 cm, depending upon the extent of surgical dissection. No discernable migration was noted when the tissue dissection was minimal. Studies utilizing a modified device equipped with electrodes indicated that migration was nominal with sufficient positional stability to ensure activation of target muscles for glottis opening. This study supported the feasibility of using a microstimulator for reanimation of paralyzed laryngeal muscles that open the airway during breathing. This innovative approach to treatment would alleviate the need for a tracheotomy or surgical resection of the vocal fold. PMID- 11499527 TI - Calibrated single-plunge bipolar electrode array for mapping myocardial vector fields in three dimensions during high-voltage transthoracic defibrillation. AB - Mapping of the myocardial scalar electric potential during defibrillation is normally performed with unipolar electrodes connected to voltage dividers and a global potential reference. Unfortunately, vector potential gradients that are calculated from these data tend to exhibit a high sensitivity to measurement errors. This paper presents a calibrated single-plunge bipolar electrode array (EA) that avoids the error sensitivity of unipolar electrodes. The EA is triaxial, uses a local potential reference, and simultaneously measures all three components of the myocardial electric field vector. An electrode spacing of approximately 500 microm allows the EA to be direct-coupled to high-input impedance, isolated, differential amplifiers and eliminates the need for voltage dividers. Calibration is performed with an electrolytic tank in which an accurately measured, uniform electric field is produced. For each EA, unique calibration matrices are determined which transform potential difference readings from the EA to orthogonal components of the electric field vector. Elements of the matrices are evaluated by least squares multiple regression analysis of data recorded during rotation of the electric field. The design of the electrolytic tank and electrode holder allows the electric field vector to be rotated globally with respect to the electrode axes. The calibration technique corrects for both field perturbation by the plunge electrode body and deviations from orthogonality of the electrode axes. A unique feature of this technique is that it eliminates the need for mechanical measurement of the electrode spacing. During calibration, only angular settings and voltages are recorded. For this study, ten EAs were calibrated and their root-mean-square (rms) errors evaluated. The mean of the vector magnitude rms errors over the set of ten EAs was 0.40% and the standard deviation 0.07%. Calibrated EAs were also tested for multisite mapping in four dogs during high-voltage transthoracic shocks. PMID- 11499528 TI - Single-unit neural recording with active microelectrode arrays. AB - This paper discusses the single-unit recording characteristics of microelectrode arrays containing on-chip signal processing circuitry. Probes buffered using on chip unity-gain operational amplifiers provide an output resistance of 200 ohm with an input-referred noise of 11-muV root-mean-square (rms) (100 Hz-10 kHz). Simultaneous in vivo recordings from single neurons using buffered and unbuffered (passive) iridium recording sites separated by less than 20 microm have shown that the use of on-chip circuitry does not significantly degrade system noise. Single-unit neural activity has also been studied using probes containing closed loop preamplifiers having a voltage gain of 40 dB and a bandwidth of 13 kHz, and several input dc-baseline stabilization techniques have been evaluated. Low-noise in vivo recordings with a multiplexed probe have been demonstrated for the first time using an external asymmetrical clock running at 200 kHz. The multiplexed system adds less than 8-muV rms of noise to the recorded signals, suppressing the 5-V clock transitions to less than 2 ppm. PMID- 11499529 TI - Dielectrophoretic trapping of dissociated fetal cortical rat neurons. AB - Recording and stimulating neuronal activity at multiple sites can be realized with planar microelectrode arrays. Efficient use of such arrays requires each site to be covered by at least one neuron. By application of dielectrophoresis (DEP), neurons can be trapped onto these sites. This study investigates negative dielectrophoretic trapping of fetal cortical rat neurons. A planar quadrupole microelectrode structure was used for the creation of a nonuniform electric field. The field was varied in amplitude (1, 3, and 5 V) and frequency (10 kHz-50 MHz). Experimental results were compared with a theoretical model to investigate the yield (the number of neurons trapped in the center of the electrode structure) with respect to time, amplitude and frequency of the field. The yield was a function of time(1/3) according to theory. However, unlike the model predicted, an amplitude-dependent frequency behavior was present and unexpected peaks occurred in the DEP-spectra above 1 MHz. Gain/phase measurements showed a rather unpredictable behavior of the electrode plate above 1 MHz, and temperature measurement showed that heating of the medium influenced the trapping effect, especially for larger amplitudes and higher frequencies. PMID- 11499530 TI - A novel method for beat-to-beat detection of ventricular late potentials. AB - A novel method for beat-to-beat detection of ventricular late potentials (VLP) from high-resolution electrocardiograms (ECGs) is presented. ECG signals from the X lead are first filtered using a bandpass filter, and then a time-sequence adaptive filter, to improve its signal-to-noise ratio. Eight features are extracted using wavelet transform, from the VLP time-frequency distribution of the filtered ECG signals, and used as inputs of specially designed artificial neural network for VLP recognition. The artificial neural network was trained and tested using clinical data, respectively. The results show that the presented method can detect beat-to-beat-based VLP with sensitivity of 80% and specificity of 77%, and the detection accuracy is 78%. PMID- 11499531 TI - Time-frequency analysis of temporomandibular joint (TMJ) clicking sounds using radially Gaussian Kernels. AB - Temporomandibular joint (TMJ) sounds and motion were recorded during two clinically-derived movements--simple jaw opening and jaw protrusion followed by opening--from ten patients. A new time-frequency method--radially Gaussian kernel distribution--was applied to classify the TMJ clicking sounds into six groups, type I to type VI, based on the time-frequency patterns of energy distribution. The number of clicks and percentage of each type were examined. Relations between the two movements were examined by the prevalence of each type. A detailed classification of TMJ clicking sounds is provided by time-frequency patterns and may provide a better understanding of temporomandibular disorders. PMID- 11499532 TI - Atrial activity enhancement by Wiener filtering using an artificial neural network. AB - This paper describes a novel technique for the cancellation of the ventricular activity for applications such as P-wave or atrial fibrillation detection. The procedure was thoroughly tested and compared with a previously published method, using quantitative measures of performance. The novel approach estimates, by means of a dynamic time delay neural network (TDNN), a time-varying, nonlinear transfer function between two ECG leads. Best results were obtained using an Elman TDNN with nine input samples and 20 neurons, employing a sigmoidal tangencial activation in the hidden layer and one linear neuron in the output stage. The method does not require a previous stage of QRS detection. The technique was quantitatively evaluated using the MIT-BIH arrhythmia database and compared with an adaptive cancellation scheme proposed in the literature. Results show the advantages of the proposed approach, and its robustness during noisy episodes and QRS morphology variations. PMID- 11499533 TI - Comments on "femoral surface strain in intact composite femurs: a custom computer analysis of the photoelastic coating technique.". PMID- 11499534 TI - Effects of low dose ultraviolet A-1 phototherapy on morphea. AB - AIM: The effects of low dose ultraviolet A-1 (UVA-1) phototherapy on different clinical stages of morphea (localized scleroderma) were analyzed in this case study. Based on these data, the different types of phototherapy described in the literature and currently used for treatment of morphea are compared. METHODS: Three patients with severe plaque type morphea in different stages were studied: one patient with late-stage lesions having stable sclerotic plaques; another patient with active inflammatory lesions; and a third patient with late-stage lesions associated with overlying lichen sclerosus et atrophicus (LSA). The treatment given was low dose UVA-1 phototherapy with single doses of 20 J/cm2 administered four times a week for 6 weeks, and once a week for another 6 weeks. RESULTS: Following UVA-1 phototherapy, the sclerotic plaques resolved, leaving smooth and soft tanned skin with normal structure, consistency and folding capability. In morphea with overlying LSA the elastic fibers did not completely return to the superficial papillary dermis despite the clinical clearance of both morphea and LSA. These data suggest that low dose UVA-1 phototherapy may improve, but not completely reverse, the histopathological changes of LSA. No side effects were observed during or after treatment. CONCLUSION: Our observations show in three patients that low-dose UVA-1 phototherapy is highly effective for treatment of all stages of morphea, including early inflammatory and late sclerotic lesions, and morphea with overlying lichen sclerosus et atrophicus. Because of its safety and efficacy, low dose UVA-1 phototherapy appears to be the treatment modality of choice. PMID- 11499535 TI - Does the minimal phototoxic dose after 8-methoxypsoralen baths correlate with the individual's skin phototype? AB - BACKGROUND/AIMS: Up to now no data have been available concerning whether there is a significant correlation between skin phototypes and the minimum phototoxic dose (MPD) after bath water delivery of 8-MOP. METHODS: The skin phototype of each of 46 patients was determined based on the individual past history of solar induced burning and tanning. In addition, the MPD of each patient was assesed after photosensitization with a warm water bath (37 degrees C, 98.6 degrees F) containing 1.0 mg/l 8-methoxypsoralen (8-MOP). Statistical analysis was performed using a Mann-Whitney U-test and Spearman rank order correlation. RESULTS: The median MPD in patients with skin phototype II was 2.0 J/cm2 (range < or =0.5 to > or =3.5) versus 1.5 J/cm2 (range 1.0 to > or =3.5) in patients with skin phototype III. There was a considerable overlap between both groups. No significant difference was detected comparing both groups (P=0.7326) and Spearman rank order correlation revealed no correlation between skin phototype and MPD. CONCLUSION: Erythemal sensitivity in PUVA bath therapy, measured as MPD, is not correlated with sun-reactive skin phototype in skin types II and III. Thus skin phototype is not a suitable indicator for the initial UVA dose in PUVA bath photochemotherapy. PMID- 11499536 TI - PUVA and PUVB in vitiligo--are they equally effective? AB - BACKGROUND/AIMS: The combination of psoralens with different types of ultraviolet (UVL) sources in the treatment of vitiligo has led to different reports of success. The purpose of this trial is to compare in a random right-left comparison study the efficacy and side effects of oral 8-MOP plus UVA (PUVA) and oral 8-MOP plus UVB (broadband, 290-320 nm P-UVB) in the treatment of vitiligo. METHODS: The study included 24 cases of extensive vitiligo involving more than 30% of the body surface area in a bilateral symmetrical distribution. Each patient received 0.7 mg/kg 8-MOP orally 2 h before the light session. The right side of the body was exposed to UVA (320-400 nm), while the left half was exposed to UVB (290-320 nm). The patients received 3 sessions/week for a total of 30 sessions. RESULTS: Both PUVA and PUVB produced moderate (50-60%) improvement, with similar incidences of phototoxic reaction and skin thickening. However, the study revealed a significant difference in the number of sessions needed to improve produce erythema and perifollicular pigmentation as well as a moderate response, the response on the UVA side always being earlier. Furthermore, the amount of joules needed to achieve the same response was 10 times greater on the UVA side than on the UVB side. CONCLUSION: The use of psoralen plus broadband UVB is as effective as PUVA in the treatment of vitiligo. However, the long-term side effects of psoralen plus UVB are unknown. PMID- 11499537 TI - A retrospective review of PUVA therapy at the National Skin Centre of Singapore. AB - BACKGROUND: Photochemotherapy (PUVA) is beneficial for the treatment of various dermatoses. The introduction of ultraviolet B and narrow-band ultraviolet B phototherapy has had a significant impact on the role of PUVA in dermatology. This study aims to assess the current role of PUVA in treating dermatoses, in a predominantly Asian population, at the National Skin Centre in Singapore. MATERIALS AND METHODS: We reviewed the clinical data of 115 patients who were started on PUVA treatment at the National Skin Centre, Singapore in 1998. We analysed the epidemiology data, the clinical response rate and the adverse effects of PUVA therapy. All of the patients continued to have ongoing maintenance treatment. RESULTS: Most of our patients were Chinese (74.8%) and male (58.3%), ranging from the ages of 4 to 74 years. Vitiligo (60.9%) was the commonest skin disorder treated with PUVA, followed by psoriasis (20.9%), endogenous eczema (11.3%), mycosis fungoides (3.5%), lichen amyloidosis (2.6%) and prurigo nodularis (0.9%). Most of our patients received paint PUVA (50.4%), 33.9% oral PUVA and 15.7% bath PUVA. The best clinical response--those patients achieving a greater that 50% clearance--was observed in patients with endogenous eczema, psoriasis and mycosis fungoides (76.8%, 73.9% and 60%, respectively). Of the vitiliginous patients, 54.3% experienced a poor response to PUVA therapy. The main side effects were mild erythema and pruritus. CONCLUSION: PUVA remains a valuable, well-tolerated therapeutic option for a variety of dermatoses. PMID- 11499538 TI - Subliminal ultraviolet-B irradiation for the prevention of vitamin D deficiency in the elderly: a feasibility study. AB - AIM: To investigate whether daily subliminal ultraviolet-B irradiation in elderly subjects can maintain a normal vitamin D status. MATERIAL AND METHODS: In an open, uncontrolled study, 10 residents (four men and six women), mean age on entry of 83 years (range 65-97 years), from a residential nursing home in County Durham, England, were exposed daily to an artificial source of ultraviolet-B radiation equivalent to 15 min of summer sunshine. The change in plasma 25 hydroxyvitamin D levels following irradiation was measured. RESULTS: Improvement in the vitamin D status of the elderly residents was achieved with no change in their routine and with no intervention on the part of carers. CONCLUSIONS: On the basis of this small pilot study we conclude that subliminal ultraviolet-B irradiation may be effective in maintaining vitamin D status in elderly people at a lower cost than that of using oral vitamin D supplementation. Further controlled studies are needed before this technique can be recommended to change health policy with regard to the vitamin D status of institutionalised elderly people. PMID- 11499539 TI - Pefloxacin and ciprofloxacin increase UVA-induced edema and immune suppression. AB - BACKGROUND: Quinolone antibiotics are popularly prescribed antibiotics because of their wide antibacterial spectrum and lowered bacterial resistance. Quinolone antibiotics are one of the well-known photosensitizers that induce phototoxicity. Their role in photocarcinogenesis has been suggested in some studies. MATERIAL AND METHODS: Mice were treated with two quinolone antibiotics (ciprofloxacin, which is less phototoxic, and pefloxacin, which is more phototoxic) to study the effect of the antibiotics on sunburn and immune suppression by ultraviolet A (UVA) irradiation. The effects of a combined treatment with UVA and these quinolone antibiotics were measured on back skin swellings, sunburn cell formations, depletion of epidermal Langerhans cells, and local and systemic suppression of contact hypersensitivity. RESULTS: Mice treated with both UVA and quinolone showed significantly increased back skin swellings and decreased epidermal Langerhans cells than mice treated with UVA only. Sunburn cells were increased significantly in mice treated with pefloxacin and 50 J/cm2 of UVA. Combination of pefloxacin and UVA suppressed local contact hypersensitivity significantly, but not systemic contact hypersensitivity. CONCLUSION: Phototoxic quinolones augmented the effect of UVA by increasing sunburn and apoptosis, depleting Langerhans cells and suppressing local immune response. By affecting apoptosis and immune suppression, they may facilitate photocarcinogenesis caused by UVA. PMID- 11499540 TI - Skin aging induced by ultraviolet exposure and tobacco smoking: evidence from epidemiological and molecular studies. AB - BACKGROUND: Tobacco smoking, similar to ultraviolet (UV) A radiation exposure, has previously been identified as an important factor contributing to premature aging of human skin. OBJECTIVE AND DESIGN: To investigate the relationship between these two environmental factors, we have conducted a cross-sectional study of 83 subjects (48 males, 35 females, age range 23-95), in which sun exposure, pack-years of smoking history and potential confounding variables were assessed by questionnaire. Facial wrinkles were quantified using the Daniell score. In order to study the molecular mechanism by which smoking caused wrinkle formation, in vitro studies were conducted to assess the alteration of matrix metalloproteinase-1 (MMP-1) mRNA expression in human fibroblasts stimulated with tobacco smoke extract or/and UVA. RESULTS: Logistic statistic analysis of the data revealed that age [odds ratio (OR)=7.5, 95% confidence interval (CI)=1.87 30.161, pack-years (OR=5.8, 95% CI=1.72-19.87), and sun exposure (OR=2.65, 95% CI=1.0-7.0) independently contributed to facial wrinkle formation. When excessive sun exposure (>2 h/day) and heavy smoking (35 pack-years) occurred together, the risk for developing wrinkles was 11.4 times higher than that of non-smokers and those with less sun exposure (<2 h/day) at the same age. The in vitro studies revealed that MMP-1 expression was significantly increased in fibroblasts after the stimulation with either tobacco smoke extract or UVA. Maximum induction was observed when cells were treated with tobacco smoke extract plus UVA, indicating that the two factors act in an additive manner. MMP-1 induction was significantly higher in the low glutathione (GSH) content fibroblast compared to that in the high GSH fibroblast, indicating that the differences in glutathione content define the susceptibility of fibroblasts towards UV- or tobacco smoking-induced MMP-1 expression. CONCLUSION: Tobacco smoke and UVA cause wrinkle formation independently of each other. We propose that both factors cause aging of human skin through additive induction of MMP-1 expression. PMID- 11499541 TI - Evaluation of the inhibitory activity of topical indomethacin, betamethasone valerate and emollients on UVL-induced inflammation by means of non-invasive measurements of the skin elasticity. AB - BACKGROUND/AIM: Topical indomethacin has been reported to inhibit ultraviolet light-induced erythema. The objective of this study was to verify this assertion and to compare indomethacin 10% ointment to beta-methasone valerate 0.1% ointment, water-in-oil emulsion and oil-in-water emulsion by means of non invasive skin elasticity measurements. METHODS: Products were applied on the back skin 60 min and 5 min before and 5 min after UVL irradiation. Untreated test sites served as controls. Clinical evaluation, measurements of epidermal hydration (Corneometer) and mechanical properties of the skin (Cutometer) were made 1 h before and 24 h after exposure. RESULTS: Test areas treated with indomethacin 60 min and 5 min before irradiation showed the significantly lowest visual erythema scores and no significant changes in skin mechanical parameters. At all other test sites, a significant decrease in elasticity parameters (Ue, Ur, Ua/Uf, Ur/Uf) and an increase in viscoelasticity parameters (Uv, Uv/Ue) of the skin were observed. No significant changes of epidermal hydration were found at any of the test sites. CONCLUSION: The inhibitory action of topical indomethacin on UVL-induced inflammation is superior to beta-methasone valerate and emollients. Non-invasive measurement of skin elasticity could be used as a supplementary tool for objective evaluation and comparison of the photoprotective activity of different topical agents. PMID- 11499542 TI - Phototesting based on a divergent beam--a study on normal subjects. AB - In a previous publication from our group, phototesting based on a single exposure to a divergent UVB beam with radially decreasing irradiance values was suggested. The aim of the present study was to evaluate technical, practical and biological aspects of the suggested method in normal subjects. Twenty healthy volunteers were provoked on the back with both a collimated beam (four fixed doses, in circular areas with a diameter of 1.5 cm) and the divergent beam (a continuous, radially attenuating dose spectrum covering an area with a diameter of 4.5 cm). Eleven of the subjects were subjected to double provocation with the divergent beam. Assessment was carried out at 6 and 24 h after exposure by measuring the diameter of the reactions both visually and by mapping the skin blood flow change with laser Doppler perfusion imaging (LDPI). Minimal erythemal dose (MED) was determined for both the collimated and the divergent provocation. The reaction diameters were used to decide MED by combination to a mm for mm mapped dose spectrum of the divergent beam profile. Dose-response curves were plotted using the quantitative response data of the LDPI-images against the corresponding dosimetry data. No systematic difference could be proven between LDPI and visual diameters and a 95% confidence interval for the mean difference was calculated to (-0.8, 2.0). Slightly greater diameters were found at the visual assessment performed at 6 h compared to 24 h (95% confidence interval (-0.1, 2.8)). Double provocation showed a good reproducibility both for the visual and the LDPI assessment (P<0.05). The divergent beam provocation allowed a more detailed discrimination of MED compared to the collimated beam provocation. The MED values determined with the divergent beam were, however, generally higher, especially in the lower range of MED values. Technical factors related to the beam divergence and the correct measurement of erythemal effective irradiance are believed to be the explanation for this phenomenon, which is thus correctable. In conclusion, the results from this study support our belief that the phototesting protocol based on a divergent beam constitutes a good opportunity for improved phototesting, since MED and dose-response characteristics may be extracted in more detail from a single UV exposure. PMID- 11499543 TI - A new method of phototesting before narrowband UVB therapy. AB - Narrowband UVB therapy is versatile and generally preferred to broadband sources. Recently, a convenient device has been described which gives 10 different doses simultaneously to the forearm to determine the minimal erythema dose (MED). It is known there is no relation between sun reactive skin type and MED. Twenty-four individuals were tested using 26% increments giving a range of MED of between 0.38 J/cm2 and over 1.5 J/cm2. When compared to conventional window testing in 15 cases, it was found to be identical or within one increment. PMID- 11499544 TI - Sunscreen protection in the ultraviolet A region: how to measure effectiveness. PMID- 11499545 TI - Growth factor insensitivity in renal failure. AB - Advanced chronic renal failure is associated with multiple endocrine and metabolic abnormalities that result from changes in the secretion and metabolism of hormones and growth factors and the target organ sensitivity to their physiological actions. As a consequence, growth retardation, bone disease, pertubations in lipid, carbohydrate and protein metabolism are commonly seen in patients with chronic renal failure. The recent availability of recombinant growth factors has provided new therapeutic opportunities for correcting these abnormalities. However because of the presence of end-organ resistance relatively high dose therapy is required and this carries an increased risk of side effects. One logical approach to this problem would be to prevent or treat the underlying resistance and thus restore sensitivity to endogenous GH or low doses of the recombinant molecule. To achieve this goal, a better understanding of the mechanism of growth factor resistance is required. In this lecture, in honor of the memory of Frank Carone. I review our current state of knowledge of the impact of advanced renal failure on the tissue sensitivity to insulin, growth hormone and insulin-like-growth factor I. PMID- 11499546 TI - Status of glucose transporters in the mammalian kidney and renal development. AB - Glucose is the main source or energy for the mammalian cells and its entry is mediated via various transporters. About 7 facilitative (GULT-1 to -7) and 2 concentrative glucose transporters (SGLT-1 and -2) have been identified. The facilitative glucose transporters allow the glucose entry into the cell interior due to the concentration gradient and the latter via the Na+-dependent electrochemical gradient. They have similar structural motifs with 12-14 putative transmembrane domains with a predicted protein size varying from 50 to 76kDa. Some of the facilitative glucose transporters (GLUT-1, -2, -4 and -5) and both the sodium glucose co-transporters (SGLT-1 and -2) are expressed in the kidney. The transporters that are involved in the major transport of glucose in the kidney include GLUT-2 and SGLT-2. They are of high capacity and low affinity type and are expressed in the S1 segment of the proximal tubule. All the transporters expressed in the kidney are developmentally regulated. The mRNA expression of renal GLUTs is variable during the fetal and postnatal periods. On the other hand the mRNA of SGLTs increases steadily from the fetal period to maturity along with the increase in their functional activity, i.e., glucose uptake. Recent studies indicate that the SGLTs are believed to selectively regulate tubulogenesis since they are expressed in the metanephric tubules very early in the embryonic life in mammals. PMID- 11499547 TI - Relevance of aldo-keto reductase family members to the pathobiology of diabetic nephropathy and renal development. AB - Aldo-keto reductases (AKRs) are a family of monomeric oxido-reductases with molecular weight ranging from 35-40 kDa and currently includes upwards of 60 members. They are expressed in a wide variety of tissues, where they catalyze the NADPH-dependent reduction of various aliphatic and aromatic aldehydes and ketones. The functions of most of the family members are not well defined. But two members, aldehyde reductase (AKRIA) and aldose reductase (AKRIB), have been extensively studied. The latter has received the most attention since being relevant to the complications of diabetes mellitus. It is up-regulated during hyperglycemia, and at the same time there is an increased activity of the sorbitol pathway and non-enzymatic glycation of proteins with ensuing damage in various tissues. It is developmentally regulated in the ocular lens, and is believed to modulate lens fiber morphogenesis during fetal life. Unlike the other AKR family members that are ubiquitously expressed, recently a renal-specific oxio-reductase has been described that is expressed exclusively in the proximal tubules. Although, it has no homology with other AKR members, it binds to NADPH with high affinity and is up-regulated in streptozotocin-induced diabetes in mice. It is also developmentally regulated and seems to selectively modulate renal tubulogenesis during embryonic life. PMID- 11499548 TI - Expression of cyclooxygenase-2 in the macula densa of human kidney in hypertension, congestive heart failure, and diabetic nephropathy. AB - Cyclooxygenase-2 (COX-2) is constitutively expressed in the macula densa of several laboratory animal species where it is considered to play a physiologic role in the regulation of basal renal function. Pertubations to normal homeostasis is shown to be associated with the upregulation of COX-2 in the macula densa of rats and dogs. In contrast, COX-2 has not been detected in the macula densa of normal adult human and non-human primate kidneys, suggesting a less prominent role of this isoform in normal renal function in these species. In this study, we characterized COX-2 expression in human kidneys collected from subjects with a clinical history indicative of compromised renal function associated with diabetic nephropathy (DN), hypertension, and congestive heart failure (CHF). COX-2 expression was evaluated by immunohistochemistry using isoform-specific antibodies and in situ hybridization. No COX-2 protein or mRNA was observed in the macula densa of normal kidneys (n= 11), whereas slight to moderate COX-2 expression was present in the macula densa of 7/15 subjects (46%) with DN, 5/11 (46%) subjects with hypertension, and 3/10 subjects (30%) with CHF. These results indicate that COX-2 is variably induced in the macula densa of the human kidney in compromised renal conditions and that COX-2-mediated prostaglandins may be involved in maintaining adequate renal functions in some patients with DN, hypertension, and CHF. This variability may be related to individual clinical status or synthesis of vasodilatory prostaglandins by cyclooxygenase-1 (COX-1). PMID- 11499549 TI - P-glycoprotein in HK-2 proximal tubule cell line. AB - P-glycoprotein (PGP) is an efflux pump physiologically expressed in the apical membrane of the proximal tubular cells. PGP may play a role in the elimination of exogenous substances such as chemotherapeutic drugs, calcium channel blockers and immunosuppressors. The involvement of renal PGP in the transport of endogenous substrates is under investigation. HK-2 is an immortalized proximal tubule cell line from normal adult human kidney, reported to retain a phenotype indicative of a well-differentiated state. No data regarding expression and/or activity of PGP in this cell line are available. The aim of this study was to ascertain the usefulness of HK-2 cell line to investigate the properties and roles of PGP in proximal tubular cells. PGP expression in HK-2 cells was determined by immunoblotting analysis using the monoclonal antibody C219. The activity of PGP was assessed by measuring the transport of the fluorescent probe Rhodamine 123 (R 123) in intact cell monostrates. The interactions of putative PGP modulators, including verapamil and cyclosporin A were also evaluated. Western blot revealed a C219 immunoreactive band of about 150 kDa consistent with the presence of PGP. HK-2 cells preloaded with R-123 rapidly effluxed the dye, the efflux being inhibited by verapamil. Verapamil and, to a major extent cyclosporin A, significantly increased R-123 intracellular accumulation. PGP immunoblottable amount was increased when cells were cultured in the presence of either cyclosporin A or dexamethasone. The results suggest that the HK-2 cells, among the various differentiation features of proximal tubules, retain also the expression of a functional PGP in their membranes and that both PGP activity and expression may be modulated by drugs. Therefore, HK-2 line appears a suitable and promising tool for the study in vitro of renal transport processes dependent on PGP. PMID- 11499550 TI - Paradoxical body and kidney growth in potassium deficiency. AB - In the growing animal, K deficiency (KD) retards body growth, but paradoxically stimulates renal growth. If KD persists, interstitial infiltrates appear and eventually tubulointerstitial fibrosis develops. In patients with chronic KD, renal cysts may form and with time tubulointerstitial disease with renal failure develops. Since early in KD, kidney IGF-I levels increase and may be a cause of the renal hypertrophy, and as TGF-beta promotes hypertrophy and fibrosis, we examined the expression of these growth factors in chronic KD. Rats were given a KD diet or pair or ad-lib fed a normal K diet. After 21 days, KD rats weighed less than pair fed controls, while the kidneys were 49% larger Serum IGF-I and kidney IGF-I protein levels were depressed, as were IGF-I mRNA levels, and is largely attributable to decreased food intake. Kidney IGFBP-1 and TGF-beta mRNA levels were increased (p < 0.05). There was marked hypertrophy and adenomatous hyperplasia of outer medullary collecting ducts, hypertrophy of thick ascending limbs of Henle (TALH) and interstitial infiltrates. Both nephron segments stained strongly for IGF-I and IGFBP-1. Only the non-hyperplastic TALH was strongly TGF beta positive. Interstitial infiltrates containing monocytes/macrophages were prominent. These findings are consistent with a sustained role for IGF-I in promoting the renal hypertrophy of KD and appear to be caused by local trapping of IGF-I by the over-expressed IGFBP-1. Localization of TGF-beta to the hypertrophied non-hypoplastic tubules containing IGF-I, suggests that TGF-beta may be acting to convert the proliferative action of IGF-I into a hypertrophic response. TGF-beta may also contribute to the genesis of the tubulointerstitial infiltrate. Finally, the reduced levels of serum IGF-1 levels may be a cause of the blunted body growth. PMID- 11499551 TI - The return of glomerular filtered albumin to the rat renal vein--the albumin retrieval pathway. AB - BACKGROUND: Recent studies have demonstrated that the normal glomerular capillary wall (GCW) is not charge selective to albumin. This means that albumin flux across the GCW is high. This has been confirmed in studies where albumin uptake by the tubules has been inhibited. Therefore, there must be a high capacity postglomerular retrieval pathway in normal kidneys that returns filtered albumin back to the blood supply. METHODS: This study identifies the presence of glomerular filtered albumin in the renal vein from the analysis of the decrease of radioactivity in the venous effluent after the injection of a pulse of tritium labeled albumin into the renal artery in vivo and in the isolated perfused kidney (IPK). RESULTS: The glomerular filtered albumin is returned to the blood supply by a high capacity pathway that transports this albumin at a rate of 1830+/-292 microg/min rat kidney (n= 14) (mean+/-SEM). This pathway has been identified under physiological conditions in vivo and in the IPK. The pathway is specific for albumin as it does not occur for horseradish peroxidase (HRP). The pathway is inhibited in a non-filtering kidney. The pathway is also inhibited by NH4Cl, an inhibitor of protein uptake. CONCLUSIONS: The high capacity retrieval pathway for albumin is most likely associated with transtubular cell transport. It is also apparent that most albuminuric states could be accounted for by the malfunctioning of this pathway without resorting to any change in glomerular permselectivity. PMID- 11499552 TI - The "fixed" charge of glomerular capillary wall as determinant of permselectivity. AB - The determinants of glomerular capillary wall (GCW) permeability to proteins have been subject of controversial discussion. To study this question we have developed a modified isolated perfused rat kidney model in which tubular transport processes are completely blocked by perfusion fixation with glutaraldehyde. This model allows to directly titrate the charge density of the GCW using albumin solutions buffered over a wide pH-range, a manipulation that cannot be performed in the intact kidney. Analyzing the results of these experiments helped to determine a fixed charge density of the GCW of 43 mEq/L. In the present work, we used the isolated perfused fixed rat kidney model to study the influence of this fixed charge on the transglomerular passage of proteins. To do this, the fixed kidney was perfused with albumin solutions containing different isoforms of horseradish peroxidase. The lowest sieving coefficient was obtained with the acidic isoform (0.035+/-0.008, n = 7), while the isoforms at pI 6.85 and 8.45 showed higher sieving coefficients (0.059+/-0.008, n = 7 and 0.090+/-0.008, n = 4, respectively). The highest sieving coefficient (0.59+/ 0.031, n = 6) was observed in perfusion experiments of the fixed kidney with cationic HRP (pI > or = 9.30). However, when comparing the sieving coefficients, the highly cationic isoform was excluded because it has a lower molecular weight than the other isoforms. The sieving coefficients of the other isoforms were significantly different (p < 0.05. ANOVA, Scheffe test). In conclusion, the presence of a discrete (even if lower than previously thought) "fixed" charge on the GCW of 43 mEq/L restricts the transglomerular passage of isoforms of horseradish peroxidase by a factor 2-3. These results imply that the influence of charge selectivity has been overstated in the literature. PMID- 11499553 TI - Influence of diuresis on enzymuria. AB - Urinary excretion of renal brush border enzymes may serve as an early marker of renal injury. However, the distinction between physiological and pathological levels remains controversial, since enzymuria is affected by physiological parameters. To clarify the influence of diuresis, we investigated the urinary excretion of alanine-aminopeptidase (AAP; EC 3.4.11.2) as function of diuretic state. 17 healthy volunteers of both sexes were subjected to protocols with sudden or prolonged water load preceded and followed by a thirst period. Urinary excretion of AAP was measured using an enzyme kinetic assay. As expected AAP excretion increased with urine flow, the increments diminished yielding an overall excretion pattern that resembled saturation kinetics. This function is described by a mathematical model. This model assumes, that AAP is released in proximal tubules at a constant rate and reabsorbed or inactivated in the distal tubule and collecting duct. Non-linear fits of the model equation to our data allowed two parameters, chi and mu, to be defined. Chi describes the rate of AAP release independent of urinary flow, and mu the ratio of distal tubular reabsorption or inactivation. If a substrate is not reabsorbed at all, mu approximates zero. Since mu fitted for AAP differed significantly from zero, this indicates reabsorption or inactivation of AAP in the distal nephron. Therefore, our study supports the theory of flow-dependent reabsorption or inactivation of AAP in the distal nephron. PMID- 11499554 TI - Renal effects of cardiac angiography with different low-osmolar contrast media. AB - The aim of this study was to evaluate the renal effects of cardiac angiography performed with three low-osmolar contrast media (CM): iopromide (IPR), ioversol (IVR) and ioxaglate (IOX). IPR and IVR are non-ionic CM, IOX is an ionic CM. Different parameters of renal function were determined before and 6, 24, 48, 72 hrs after angiography in 45 patients: 15 patients were examined with IPR, 15 with IVR and 15 with IOX. Glomerular effects--Plasma creatinine increased slightly at the 24th hour after IVR and IOX and at 48 hours after IOP. A significant increase in plasma beta2-microglobulin was observed, at the same time, only after IOX. A significant decrease in creatinine clearance was found at 6 hours after IOX. No significant variations in glomerular filtration rate (GFR) and in effective renal plasma flow were found at 48 hours after cardiac angiography; while filtration fraction was significantly reduced after IOP and IOX. Tubular effects--A marked decrease in sodium clearance and a relevant increase of urinary activities of different tubular enzymes were found after cardiac angiography with all CM, but were more evident after the ionic CM IOX, than after the two non-ionic agents. These tubular effects reached the maximum between 6 and 24 hours and returned to baseline within 72 hrs after cardiac angiography. In conclusion, slight glomerular effects were observed mainly after IOX. A reversible tubular malfunction was found with the three low-osmolar CM and was more evident after ionic CM IOX. thus suggesting that other mechanisms, besides osmolarity, play a role in tubular toxicity due to CM. In no patient did the glomerular and tubular effects of CM have a clinical relevance. PMID- 11499555 TI - Potentials and limitations of the low-molecular-weight protein lysozyme as a carrier for renal drug targeting. AB - Selective targeting of drugs to the kidney may enable an increased renal effectiveness combined with a reduction of extrarenal toxicity. Intrarenal delivery to the proximal tubular cell can be achieved using low-molecular-weight proteins, such as lysozyme. Administration of high dosages of lysozyme, required to study the effects of such conjugates in vivo, however, is restricted since a partial escape of the renal reabsorption and the occurrence of unwanted effects on systemic blood pressure and renal function may occur. The purpose of this study was to investigate the optimal parenteral administration schedule and the maximum dose of lysozyme, providing the most optimal tubular reabsorption and at the same time a minimal effect on blood pressure and renal hemodynamics, comparing continuous infusion of lysozyme with single dose injections. Urinary lysozyme excretion increased dose-dependently, both during continuous infusion and intravenous bolus injections. However, this loss of intact lysozyme into the urine was much higher after 3 injections of in total 250 mg x kg(-1) x 6 h(-1) (51.8+/-3.7% of the dose) compared to the same dose administered by continuous infusion (11.7+/-2.4%, P < 0.001). Continuous infusion of lysozyme up to 1000 mg x kg(-1) in 6 hours had no effect on systemic blood pressure, whereas a bolus injection of lysozyme (167 mg x kg(-1)) resulted in reversible blood pressure lowering of 52.2+/-2.2% (P<0.001). A dose-dependent decline of the glomerular filtration rate was observed at dosages of lysozyme higher than 100 mg x kg(-1) x 6 h(-1), with a maximal reduction of 53.0+/-3.7% after infusion of 1000 mg x kg( 1) x 6 h(-1). Effective renal plasma flow was less affected and only lowered statistically significant at dosages of 500 (-12.6+/-3.3%, P<0.05) to 1000 mg x kg(-1) x 6 h(-1) (-17.2+/-3.9%, P<0.01). We conclude that bolus injections of lysozyme should not be used for renal targeting purposes since it results in considerable tubular loss of lysozyme in the urine as well as cardiovascular side effects. In contrast, continuous infusion of lysozyme using dosages sufficient for renal drug targeting (maximally 15 mg x kg(-1) x h(-1)) only has minimal effects on blood pressure and renal hemodynamics, with a minimal urinary lysozyme loss as well. PMID- 11499556 TI - Blood tubing and cytokine production: effect of sterilization. AB - Blood tubings commonly represent an integral component of hemodialysis circuits. Different factors may influence their biocompatibility, such as the type of material, the sterilization mode and the geometry. In vivo the final biocompatibility may be further complicated by the individual host response, the flow parameters, and the impact of mechanical trauma on blood's cellular components (i.e. erythrocytes). In this in vitro study we evaluated some commercially available blood tubings sterilized by different methods as to their interaction with normal leukocyte population and tested the response of these cells in terms of cytokines (IL-1beta, IL-1Ra, TNF-alpha). As a positive control, leukocytes were incubated with 0.5 ng/mL of bacterial lipopolysaccharide (LPS) or with Cuprophan of comparable surface. The results showed that cytokine production was markedly reduced, particularly in the case of gamma-ray-sterilized tubings. Of interest, it was not always related to the adherence. However in some cases, particularly of gamma-ray sterilization, adherence was none, despite the cytokine production. PMID- 11499557 TI - Reappraisal of serum beta2-microglobulin as marker of GFR. AB - INTRODUCTION: Beta 2 microglobulin (beta2M) is filtered by the glomeruli and reabsorbed by the proximal tubular cells where it is metabolized. Its plasma concentration increases with decreasing renal function. AIM: To compare serum creatinine (Cr) and serum beta2M as markers of GFR. PATIENTS AND METHODS: In 160 adult patients, with various kidney diseases and different GFR, serum Cr (autoanalyzer), serum beta2M (RIA) and GFR (bladder cumulative method using 99mTc DTPA as glomerular tracer) were measured in the same day. RESULTS: A linear relationship was observed between In GFR and both In serum Cr (lnCr=3.112-0.716 lnGFR; r=0.92) and ln serum beta2M (lnbeta2M= 4.274-0.814 lnGFR; r = 0.90). With decreasing GFR the increase in serum beta2M was higher than that of serum Cr (see regression coefficients that are significantly different). The normal upper limit of serum Cr corresponds to a GFR 48.1 mL/min while that of serum beta2M to a GFR 65.0. With decreasing GFR the increase of serum beta2M occurs before than that of serum Cr. CONCLUSIONS: With declining renal function, serum beta2M increases more and before than serum Cr. Serum beta2M is a good endogenous marker of GFR, better than serum Cr. PMID- 11499558 TI - Cystatin C in a rat model of end-stage renal failure. AB - BACKGROUND: Cystatin C (MW 13kDa) serum concentration reflects glomerular filtration rate better than creatinine. Like other low-molecular weight proteins it is not eliminated by dialysis. Still, cystatin C serum concentrations do not rise progressively in end-stage renal failure and rarely exceed 10 mg/L (i.e. 8 times the upper limit of normal). OBJECTIVE: To study cystatin C kinetics in a rat model of end-stage renal failure. METHODS: Sequential bilateral nephrectomy was performed seven days apart in 13 male Sprague-Dawley rats as described by Levine and Saltzman. Serum cystatin C (Cystatin C PET-kit, DAKO), creatinine and total protein were measured in daily intervals after the second nephrectomy. Linearity of the anti-human cystatin C assay for rat cystatin C was tested using dilutions of uremic rat serum. Rats were sacrificed for signs of severe uremia on days 10 (n=5), 11 (n=4) and 12 (n = 5). RESULTS: At baseline, mean (+/- SE) cystatin C was 1.59+/-0.041 mg/L, creatinine 19.6+/-1.2 micromol/L. Following bilateral nephrectomy, cystatin C immediately rose to 3.82+/-0.15 mg/L, creatinine to 312+/-20 micromol/L. During the following days, cystatin C concentration stabilized to 4 mg/L approximately whereas creatinine continued to rise to 822+/-185 kmol/L on day 12. Correction for the decrease in serum total protein concentration from 48.9+/-2.3 g/L to 37.4+/-3.6 g/L did not alter these results. CONCLUSION: The kinetics of cystatin C and creatinine in this rat model of end-stage renal failure are in accordance with human data suggesting a change in cystatin C production or extra-renal elimination in severe chronic uremia. PMID- 11499559 TI - Assessment of renal function in renal transplant patients using cystatin C. A comparison to other renal function markers and estimates. AB - To date, little evidence is available to define the role of cystatin C in patients with renal transplants. Thus, to assess, whether cystatin C (CysC) provides better information on renal function than other markers, CysC, creatinine clearance (CrCl), serum creatinine (SCr), beta2-microglobulin (beta2 M), and 125I-Iothalamate clearance were determined in 30 patients. Correlation and ROC curves were obtained and characteristics like sensitivity and specificity were calculated. Further, to evaluate the usefulness of these markers for monitoring, intraindividual coefficients of variation for CysC and SCr measurements were compared in 85 renal transplant patients. CysC correlated best with GFR, whereas SCr, CrCl and beta2-M all had lower correlation coefficients. CysC was superior to SCr, even when renal function equations of were used. The diagnostic accuracy of CysC was significantly better than SCr. but did not differ significantly from CrCl and beta2-M. Together, our data show that in patients with renal transplants, CysC has a similar diagnostic value as CrCl. However, it is superior to determinations of SCr. The intraindividual variation of CysC is significantly greater than that of SCr. This might be due to better ability of CysC to reflect temporary changes especially in mildly impaired GFR, most critical for early detection of rejection and other function impairment. In conclusion, CysC allows for easy and accurate assessment of renal function (GFR) in steady state renal transplant patients and is clearly superior to the commonly used serum creatinine. PMID- 11499560 TI - Relationship between renal function and blood level of chromogranin A. AB - Chromogranin A (CGA) is a low MW (49,000) acidic hydrophilic protein. It is synthesized in the chromaffm granules of the neuroendocrine cells, and has been found circulating in the blood of healthy subjects. The aim of this study was to assess the relationship between serum levels of CGA and renal function. One hundred two renal patients (45 M and 57 F; age 14-76 years, mean 52) participated in the study. Glomerular filtration rate (GFR) was measured by the bladder cumulative method, using 99mTc-DTPA as a tracer. Blood CGA was determined by RIA. Plasma creatinine, beta2microglobulin (beta2m) and tumor associated trypsin inhibitor (TATI) were also determined. The reduction in renal function was associated with an increase in all of the above studied parameters. In patients with advanced renal failure (GFR <20 mL/min) CGA levels increased by 22-fold as compared to the patients with normal renal function (GFR> 100 mL/min). The other studied parameters were also increased but to a lesser degree, e.g., TATI 14-, beta2m 8- and creatinine 5-fold. The results of this study demonstrate that renal handling of the CGA is similar to other low MW proteins, and it accumulates in the blood in renal failure. PMID- 11499561 TI - Connective tissue growth factor and its regulation: a new element in diabetic glomerulosclerosis. AB - Connective tissue growth factor (CTGF), a member of the closely related CCN family of cytokines appears to be fibrotic in skin. To determine whether CTGF is implicated in diabetic glomerulosclerosis we studied cultured rat mesangial cells (MC) as well as kidney cortex and microdissected glomeruli from obese, diabetic db/db mice and their normal counterparts. Exposure of MC to rhCTGF significantly increased fibronectin and collagen type I secretion. Further, unstimulated MC expressed low levels of CTGF message and secreted minimal amounts of CTGF protein (36-38 kDa). However, exposure to TGF-beta, increased glucose concentrations, or cyclic mechanical strain, all causal factors in glomerulosclerosis, markedly induced the expression of CTGF transcripts. With all but mechanical strain there was a concomitant stimulation of CTGF protein secretion. TGF-beta also induced abundant quantities of a small molecular weight form of CTGF (18 kDa). The induction of CTGF protein by a high glucose concentration was mediated by TGF beta, since a TGF-beta neutralizing antibody blocked this stimulation. In vivo studies using quantitative RT-PCR demonstrated that while CTGF transcripts were low in the glomeruli of control mice, expression was increased 27-fold after approximately 3.5 months of diabetes. These changes occurred early in diabetic nephropathy when mesangial expansion was mild, and interstitial disease and proteinuria were absent. A substantially reduced elevation of CTGF mRNA (2-fold) observed in whole kidney cortices indicted that the primary alteration of CTGF expression was in the glomerulus. These results suggest that CTGF upregulation is an important factor in the pathogenesis of mesangial matrix accumulation in both diabetic and non-diabetic glomerulosclerosis, acting downstream of TGF-beta. PMID- 11499562 TI - The key role of the transforming growth factor-beta system in the pathogenesis of diabetic nephropathy. AB - Progressive renal injury in diabetes mellitus leads to major morbidity and mortality. The manifestations of diabetic nephropathy may be a consequence of the actions of certain cytokines and growth factors. Prominent among these is transforming growth factor-beta (TGF-beta) because it promotes renal cell hypertrophy and stimulates extracellular matrix accumulation, the two hallmarks of diabetic renal disease. In cell culture, high ambient glucose increases TGF beta mRNA and protein in proximal tubular, glomerular epithelial, and mesangial cells. Neutralizing anti-TGF-beta antibodies prevent the hypertrophic and matrix stimulatory effects of high glucose in these cells. In experimental and human diabetes mellitus, several reports describe overexpression of TGF-beta in the glomeruli and tubulointerstitium. We demonstrate that short-term treatment of diabetic mice with neutralizing monoclonal antibodies against TGF-beta significantly reduces kidney weight and glomerular hypertrophy and attenuates the increase in extracellular matrix mRNAs. Long-term treatment of diabetic mice further improves the renal pathology and also ameliorates the functional abnormalities of diabetic nephropathy. Finally, we provide evidence that the renal TGF-beta system is significantly up-regulated in human diabetes. The kidney of a diabetic patient actually elaborates TGF-beta1 protein into the circulation whereas the kidney of a non-diabetic subject extracts TGF-beta1 from the circulation. The data we review here strongly support the hypothesis that elevated production or activity of the TGF-beta system mediates diabetic renal hypertrophy and extracellular matrix expansion. PMID- 11499563 TI - Vascular endothelial growth factor (VEGF) and VEGF receptors in diabetic nephropathy: expression studies in biopsies of type 2 diabetic patients. AB - Vascular endothelial growth factor (VEGF) is involved in the pathogenesis of diabetic retinopathy but its role in diabetic nephropathy is only speculative so far. It has been shown that in renal cortex of normal kidneys, glomerular and tubular epithelial cells express VEGF and that VEGF 165 is the predominant isoform. Two VEGF receptors, KDR (kinase domain region) and Flt-1 (fms-like tyrosine kinase) are co-expressed by glomerular and peritubular capillary endothelial cells. However, VEGF and VEGF receptors are predominantly expressed at glomerular level. We recently demonstrated that in type 2 diabetic patients glomerular qualitative and quantitative changes of VEGF mRNA expression are associated with functional and structural renal changes. In the present work we focused on the tubulo-interstitial compartment; by reverse transcription/polymerase chain reaction (RT/PCR) we evaluated the expression of VEGF, KDR, Flt-1 and the relationship between the two main type of VEGF isoforms, VEGF121 and VEGF165 in the tubulo-interstitium of type 2 diabetic patients. Patients were divided in three category on the basis of renal structure pattern: CI, with normal or near normal renal structure; CII, with glomerular and tubulo interstitial lesions occurring in parallel (typical diabetic nephropathology); CIII, with atypical pattern of renal injury, i.e., more severe tubulo interstitial and vascular than glomerular changes. Comparison between the two cortical compartments revealed that, both in glomeruli and in tubulo interstitium. VEGF121 isoform exceed VEGF165 while Flt-1 was significantly lower in glomeruli. CIII patients had the lowest tubulo-interstitial level of VEGF and Flt-1 mRNAs. These results suggest that the transcriptional shifting from VEGF165 to VEGF121 isoform and the unbalanced FIt-1 expression between tubulo interstitium and glomeruli could be involved in the pathogenesis of diabetic nephropathy. Furthermore, at least in CIII patients, down-regulation of the VEGF Flt-1 system could be involved in the mechanisms leading to tubulointerstitial diabetic lesions. PMID- 11499564 TI - Role of proteinuria reduction in the progression of IgA nephropathy. AB - Proteinuria has been shown to play a causal role in the progression towards ESRD of IgA nephropathy (IgAN). We demonstrated that steroids are effective in reducing proteinuria and preserving renal function. AIM: to evaluate the long term effect of steroids in IgAN patients (6th year evaluation) and better clarify the role of proteinuria reduction in slowing down the progression. METHODS: multicenter randomized controlled trial of 86 adult IgAN patients with serum creatinine < or = 1.5 mg/ dL and moderate proteinuria. They received either supportive therapy or methylprednisolone 1-g i.v. for three days at months 1, 3, and 5, plus oral prednisone (0.5 mg/kg every other day for six months). RESULTS: Proteinuria significantly decreased in the treated patients (from 2.0+/-0.60 g/24 h at baseline to 1.0+/-0.68 g/24 h at six months) and remained stable till the 6th year (0.67+/-0.5 g/24 h), it slightly decreased in the control group. Six year renal survival was significantly better in the steroid than in the control group: 9 patient (20.9%) in the steroid group and 15 (34.8%) in the control group reached the primary end-point of a 50% increase in serum creatinine from baseline. Five controls and none of the steroid-treated patients started dialysis. Steroid-treated patients did not experience any major side effects during follow-up. CONCLUSIONS: Steroids significantly reduce proteinuria and protect against renal function deterioration in IgAN patients. Early reduction of proteinuria could also be marker of a persistent reduction in its levels over time and of a better outcome in the long term. PMID- 11499565 TI - Uninephrectomy increases kidney beta2-microglobulin: can it play a role in the progression of kidney damage? AB - Beta2-microglobulin (beta2M) is highly accumulated by the kidneys of normal rats. The aim of this study was to verify if uninephrectomy can modify the renal uptake of labeled beta2M. For this purpose the radioactivity of plasma and those of the remaining kidney, liver and urine have been measured in uninephrectomized rats (NX) and in controls (C) at different times after the injection as i.v. bolus of 131I-beta2M. The experiments were performed in 114 Sprague-Dawley male rats. Fifty seven animals underwent right nephrectomy, the other animals being the C. NX and their C were divided in 3 groups, studied 2, 4 and 6 weeks after nephrectomy, respectively. Part of the animals were sacrificed 12 min after the injection of labeled beta2M (peak-time, i.e. time of highest kidney accumulation of 131I-beta2M in the normal rat) and part 10 min later. The results demonstrate that: - uninephrectomy increases plasma retention of 131I-beta2M - kidney uptake (total and per gram) is always higher in NX - liver uptake (much lower than that of kidney) is not influenced by uninephrectomy - urine excretion of radioactivity is minimal in both NX and C. The behavior of beta2M is similar to that we previously observed with alpha1-microglobulin and lysozyme. The higher kidney content of some low mw proteins after uninephrectomy could play a role in the progressive reduction of renal function determined by the reduction of renal mass. PMID- 11499566 TI - Renal tubular peptide catabolism in chronic vascular rejection. AB - Chronic vascular rejection (CR) is the commonest cause of renal transplant loss, with few clues to etiology, but proteinuria is a common feature. In diseased native kidneys, proteinuria and progression to failure are linked. We proposed a pathogenic role for this excess protein at a tubular level in kidney diseases of dissimilar origin. We demonstrated in both nephrotic patients with normal function and in those with failing kidneys increased renal tubular catabolism and turnover rates of a peptide marker, Aprotinin (Apr), linked to increased ammonia excretion and tubular injury. These potentially injurious processes were suppressed by reducing proteinuria with Lisinopril. Do similar mechanisms of renal injury and such a linkage also occur in proteinuric transplanted patients with CR, and if so, is Lisinopril then of beneficial value? We now examine these aspects in 11 patients with moderate/severe renal impairment (51CrEDTA clearance 26.2+/-3.3 mL/min/1.73 m2), proteinuria (6.1+/-1.5 g/24 h) and biopsy proven CR. Lisinopril (10-40 mg) was given daily for 2 months in 7 patients. Four others were given oral sodium bicarbonate (Na HCO3) for 2 months before adding Lisinopril. Renal tubular catabolism of intravenous 99mTc-Apr (Apr* 0.5 mg, 80MBq), was measured before and after Lisinopril by gamma-ray renal imaging and urinary radioactivity of the free radiolabel over 26 h. Fractional degradation was calculated from these data. Total 24 h urinary N-acetyl-beta-glucoaminidase (NAG) and ammonia excretion in fresh timed urine collections were also measured every two weeks from two months before treatment. After Lisinopril proteinuria fell significantly (from 7.8+/-2.2 to 3.4+/-1.9 g/24 h, p<0.05). This was associated with a reduction in metabolism of Apr* over 26 h (from 0.5+/-0.05 to 0.3+/-0.005% dose/h, p < 0.02), and in fractional degradation (from 0.04+/-0.009 to 0.02+/-0.005/h, p<0.01). Urinary ammonia fell, but surprisingly not significantly and this was explained by the increased clinical acidosis after Lisinopril, (plasma bicarbonate fell from 19.1+/-0.7 to 17.4+/-0.8 mmol/L, p < 0.01), an original observation. Total urinary NAG did fall significantly from a median of 2108 (range 1044-3816) to 1008 (76-2147) micromol/L, p < 0.05. There was no significant change in blood pressure or in measurements of glomerular hemodynamics. In the 4 patients who were given Na HCO3 before adding Lisinopril, both acidosis (and hyperkalemia) were reversed and neither recurred after adding Lisinopril. These observations in proteinuric transplanted patients after Lisinopril treatment have not been previously described. PMID- 11499567 TI - Proteinuria in focal segmental glomerulosclerosis: role of circulating factors and therapeutic approach. AB - The clinical course of primary Focal Segmental Glomerulosclerosis (FSGS) is frequently complicated by nephrotic range proteinuria and progression to renal failure. The high recurrence rate of the disease in transplanted kidney suggests the hypothesis that such patients have a circulating factor that alters glomerular capillary permeability. In recent years some authors found that serum from patients with FSGS increases glomerular permeability to albumin and partially identified the permeability factor (PF) as a protein of 30-50 Kd m.w. The removal of this protein by means of Plasma Exchange (PE) or plasma Immunoadsorption by Protein A (IA) decreased proteinuria. In this report we provide preliminary data about the prevalence of PF and the therapeutic effect of its removal by IA, in 3 pts with recurrence in the transplanted kidney, and 4 with FSGS of the native kidneys. They were resistant to corticosteroids (CS) and immunosuppressive (IS) therapy. 10 IA sessions were performed in 4 weeks: if a remission was achieved IA was gradually tapered. The level of PF in the serum was measured by an in vitro assay to determine the glomerular permeability to albumin. The FSGS was histologically proven in all cases and the degree of evolution was evaluated. PF levels, serum creatinine, daily proteinuria and serum albumin were monitored. The 3 patients with recurrent FSGS had a normalization of the PF levels; 2 had a clinical remission. In FSGS of native kidneys PF was elevated in 3/4 cases; 1 had a clinical remission; 2 with extensive sclerohyalinosis and 1 without PF levels did not improve. Our results confirm that most patients with FSGS have high PF serum levels and suggest that its removal can be beneficial. PMID- 11499568 TI - Myofibroblast: a prognostic marker and target cell in progressive renal disease. AB - Myofibroblasts play an important role in many tissue injuries, and particularly in renal disease. The myofibroblast differentiation is an early event in the development of fibrosis. Myofibroblast-like cells express smooth muscle (SM) cytoskeletal markers (alpha-SM actin in particular) and participate actively in the production of extracellular matrix. Identification of early risk factors, particularly histological criteria, could be useful to identify patients at risk of progressive renal failure and needing a treatment. The evaluation of myofibroblast differentiation in renal tissue may reflect the intensity of tissue injury, predict long term outcome of chronic renal disease and help physicians to select therapeutic choices. More than a disease activity indicator. myofibroblasts appear to be a pivotal target for future therapies in progressive renal disease. PMID- 11499569 TI - C reactive protein in patients with chronic renal diseases. AB - Base-line serum levels of plasma C-reactive protein (CRP) are predictive of future myocardial infarction and sudden cardiac death in apparently healthy subjects, suggesting the hypothesis that chronic inflammation might be important in the pathogenesis of atherothrombosis. CRP production is mediated by several inflammatory mediators: interleukin 6 (IL-6) is currently felt to be the major cytokine influencing the acute phase response. CRP and other acute phase proteins are elevated in dialysis patients and cardiovascular diseases represent the single largest cause of mortality in chronic renal failure patients. Little information is available, however regarding CRP and IL-6 plasma levels in pre dialysis renal failure. Plasma CRP was determined by a modification of the laser nephelometry technique; IL-6 by immunoassay (RD System); and fibrinogen, serum albumin, cholesterol, triglycerides, hematocrit, white blood cell count, erythrocytic sedimentation rate (ESR) and urinary protein levels by standard laboratory techniques. Results were obtained in 102 chronic pre-dialysis patients whose mean age was 53+/-5.8 years with a mean creatinine clearance (C(Cr)) of 52+/-37 mL/min). CRP was greater than 5 mg/L in 25% of the global population. CRP and IL-6 were 4.0+/-4.6 mg/L and 5.8+/-5.6 pg/mL, respectively and were not significantly correlated (r=0.11, p=n.s.). CRP and IL-6 were however related with renal function (CRP versus C(Cr) r=-0.40 p <0.001; IL- 6 versus C(Cr) r=-0.45; p <0.001). When patients were divided in two groups according to renal function, CRP resulted 7.4+/-6.3 mg/L in the group of patients with a C(Cr) lower than 20 mL/min (n=32) and 2.76+/-4.35 in the group of patients with a C(Cr) higher than 20 mL/min (n = 70) (p <0.0001). CRP and IL-6 were positively related with ESR (r=0.32 and 0.46 respectively). Serum albumin levels were not significantly different in the two groups of patients (3.2+/-0.4 versus 3.0+/-0.5 g/dL). CRP and serum albumin were not significantly related (r=0.17). CRP and IL-6 correlated positively with ESR (r=0.32 and 0.46 respectively). In pre-dialysis patients we have demonstrated an increase in both CRP and IL-6 that occurs as renal function decreases. These data provided evidence of the activation - even in the predialysis phase of renal failure - of mechanisms known to contribute to the enhanced cardiovascular morbidity and mortality of the uremic syndrome. PMID- 11499570 TI - Oxidation of albumin is enhanced in the presence of uremic toxins. AB - Albumin has been considered a "sacrificial plasma antioxidant" due to the high reactivity of the protein sulfhydryl groups with oxidants such as hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). Based on its large quantity and high turnover. It is considered as one of the most important plasma antioxidants for protecting key cellular and regulatory proteins. Since hemodialysis patients have lower overall levels of albumin and possible protein modifications due to uremic toxins, we investigated whether modifications by various uremic toxins would affect the susceptibility of albumin to an oxidative challenge. We incubated bovine serum albumin in the presence of carboxymethyllysine (CML) (10 micromol/L(-1) mmol/L), methyl glyoxal (50 micromol/L(-5) mmol/L), p-cresol (100 micromol/L-10 mmol/L) or hippuric acid (200 micromol/L-20 mmol/L) for 16 hours at 37 degrees C and then subsequently added 0.5 mmol/L(-1) mmol of H2O2/HOCl. We measured the extent of protein modification by the loss of protein sulfhydryl groups, dityrosine formation and the formation of advanced oxidation protein products (AOPP). Incubation of albumin with the uremic toxins caused a loss of protein sulfhydryl groups and an increase in dityrosines and AOPP. The presence of uremic toxins had no effect on the loss of protein sulfhydryl groups after addition of H2O2/HOCl; however, low levels of CML, p-cresol and methyl glyoxal inhibited the formation of AOPP and dityrosines. We suggest that uremic toxins may possibly play a role in mediating free radical initiated protein damage. PMID- 11499571 TI - Interferon alpha2b increases paracellular permeability of renal proximal tubular LLC-PK1 cells via a mitogen activated protein kinase signaling pathway. AB - The therapeutic administration of Interferon alpha2b (IFNalpha) is often accompanied by impaired renal function, i.e. reduced glomerular filtration rate and sometimes a so-called "capillary leak syndrome". To clarify the mechanism behind the renal dysfunction, confluent monolayers of LLC-PK1 cells were used as a model system to analyze the effects of IFNalpha on renal tubular epithelium. Examination of epithelial barrier function via measurement of transepithelial resistance (TER) revealed a dose dependent increase in paracellular permeability by IFNalpha treatment. The effect was reversible upon removal of IFNalpha at doses up to 5 x 10(3) U/mL. Apical or basolateral application of IFNalpha yielded the same decrease in TER. Tyrphostin A25, an inhibitor of phosphotyrosine kinases, ameliorated the IFNalpha induced decrease of TER. In order to unravel intracellular signal transduction pathways that may mediate IFNalpha induced changes of epithelial barrier function, we inhibited IFNalpha signaling through a mitogen activated protein kinase pathway by the Mek1 inhibitor PD98059. The inhibitor could be shown to prevent IFNalpha induced decrease of transepithelial resistance. Inhibitors of the p38 mitogen activated protein kinase pathway did not affect IFNalpha mediated changes of epithelial barrier function, indicating a highly specific role for the Mek/Erk pathway. Activation of mitogen activated protein kinase pathways by epidermal growth factor or anisomycin could not, per se, imitate the effect of IFNalpha on the paracellular permeability of LLC-PK1 monolayers. These findings provide evidence that IFNalpha can affect barrier function in renal epithelial cells via activation of the Mek/Erk pathway. PMID- 11499572 TI - The role of tubular cells in the progression of renal damage: guilty or innocent? PMID- 11499573 TI - Production and activation of hepatocyte growth factor in acute renal failure. AB - Hepatocyte growth factor (HGF) facilitates the regeneration of injured kidney in acute renal failure (ARF). HGF is produced as a single-chain precursor by cells of mesenchymal origin and is converted to a biologically active, heterodimeric molecule by proteolytic processing. We studied HGF mRNA and protein levels in systemic organs of glycerol-induced ARF rats, a model of crush syndrome. HGF protein concentration of tissue homogenate was measured by ELISA. Both mRNA and protein levels were increased in liver and spleen at 24 hours after the glycerol injection whereas HGF protein level was decreased in the injured kidney. Expression of HGF receptor/c-met mRNA was elevated only in the kidney. These results suggest that HGF supplied in an endocrine manner may play an important role in the regenerating process following ARF. Next, we measured serum HGF concentration by ELISA in 8 ARF patients caused by crush syndrome and the molecular size of serum HGF was determined by immunoblotting. Although serum HGF levels elevated in all patients, the HGF levels did not associate with their prognoses. While a single-chain molecule was predominantly observed in sera from chronic renal failure patients and healthy subjects, the majority of serum HGF was a heterodimeric form in 7 ARF patients. In one patient who developed disseminated intravascular coagulation syndrome and had a poor prognosis, a single-chain molecule was predominant although the serum HGF concentration was equivalent. These data suggest that the activity of proteolytic processing may be also an important factor for the expression of the biological function of HGF. PMID- 11499574 TI - Urinary epidermal growth factor in different renal conditions in children. AB - Several studies have demonstrated the important role of growth factors, particularly epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha), in cellular growth after renal damage. EGF is mainly synthesized by the kidney. Many studies indicate that urinary EGF concentration significantly decreases in patients with acute and chronic renal failure. In this study we determined urinary EGF concentrations in children with renal and/or urological pathologies. We investigated 38 patients, 17 males and 21 females, of 3.34+/-2.96 years (mean +/- standard deviation), who were followed in the Nephrologic Unit of the Pediatric Department of the University of Verona for recurrent urinary tract infections: seven of these had vesicoureteric reflux and 4 had hypodysplasia. The results were compared with those from a healthy age-matched group of 44 children. In all patients, we assessed renal function including an examination of the urine with a microbiological evaluation. Moreover, a renal ultrasound and a voiding cystourethrogram were performed. PMID- 11499575 TI - The production of poly-(gamma-glutamic acid) from microorganisms and its various applications. AB - This review article deals with the chemistry and biosynthesis of poly-(gamma glutamic acid) (gamma-PGA) produced by various strains of Bacillus. Potential applications of gamma-PGA as thickener, cryoprotectant, humectant, drug carrier, biological adhesive, flocculant, or heavy metal absorbent, etc. with biodegradability in the fields of food, cosmetics, medicine and water treatments are also reviewed. PMID- 11499576 TI - An initial assessment of spent mushroom compost as a potential energy feedstock. AB - The Irish mushroom industry has expanded rapidly in the last decade, particularly in the border counties. Its success has generated concern, however, as the production of spent mushroom compost (SMC) has increased. Until recently, SMC as an agricultural waste has been disposed of without due consideration to the environment. As County Councils increasingly address river pollution, restrictions will prevent expansion of the mushroom industry without an SMC waste management solution. This paper looks at the availability and composition of SMC and considers its potential as an energy feedstock. Variability in SMC composition was investigated by sampling from various locations over several months. Analyses showed that, on a dry ash free basis, SMC has a calorific value equivalent to sewage sludge which has been successfully fired for many years. Compositional analysis confirmed the dry fuel as consistent in make-up and showed the moisture content to vary within 60-77%, on an as-received basis. PMID- 11499577 TI - Development of a high-temperature air-blown gasification system. AB - Current status of high-temperature air-blown gasification technology development is reviewed. This advanced gasification system utilizes preheated air to convert coal and waste-derived fuels into synthetic fuel gas and value-added byproducts. A series of demonstrated, independent technologies are combined to form the core of this gasification system. A high-temperature, rapid devolatilization process is used to enhance the volatile yields from the fuel and to improve the gasification efficiency. A high-temperature pebble bed filter is used to remove to the slag and particulates from the synthetic fuel gas. Finally, a novel regenerative heater is used to supply the high-temperature air for the gasifier. Component development tests have shown that higher gasification efficiencies can be obtained at more fuel-rich operating conditions when high-temperature air is used as the gasification agent. Test results also demonstrated the flex-fuel capabilities of the gasifier design. Potential uses of this technology range from large-scale integrated gasification power plants to small-scale waste-to-energy applications. PMID- 11499578 TI - Biological formation of volatile phosphorus compounds. AB - Phosphine and phosphides are reported to occur at numerous environmental sites such as fresh and marine sediments, landfills, faecal matter, biogas digesters and soils. The concentrations are several log units lower than the time-weighted average exposure standard, i.e. in the order of ng per m3 of gas or ng per kg material. Research about the biological formation of highly reduced gaseous phosphorus compounds dates back more than a hundred years. The early reports had to deal with a lot of scepticism. Thanks to new analytical tools (gas chromatography) it has become clear, during the last decade, that phosphine is a global constituent of the atmosphere. Pure strains of micro-organisms cultivated under highly anaerobic conditions were shown to produce phosphine. Thermodynamic considerations indicate that it is very improbable that the reduction of phosphate to phosphine is endergonic. Therefore the generation of phosphine cannot be compared with sulphidogenesis and methanogenesis. There seems to be a link between the existence of highly reactive gaseous phosphorus compounds and increased levels of metal corrosion. The reactive compounds could be formed by micro-organisms or they are liberated from phosphorus-containing impurities in the iron by the action of bacterial metabolites. The biochemical pathways responsible for the production of gaseous phosphorus compounds have not been characterised yet. PMID- 11499579 TI - Fungal decolorization of dye wastewaters: a review. AB - In recent years, there has been an intensive research on fungal decolorization of dye wastewater. It is becoming a promising alternative to replace or supplement present treatment processes. This paper examines various fungi, living or dead cells, which are capable of decolorizing dye wastewaters; discusses various mechanisms involved; reports some elution and regeneration methods for fungal biomass; summarizes the present pretreatment methods for increasing the biosorption capacity of fungal biomass; discusses the effect of various factors on decolorization. PMID- 11499580 TI - Immobilized cell technology applied in solubilization of insoluble inorganic (rock) phosphates and P plant acquisition. AB - This paper reviews current knowledge of the production of organic acids by immobilized microorganisms with a simultaneous solubilization of rock phosphate in fermentation and soil conditions. The most widely applied methods are based on the passive immobilization in preformed porous carriers and entrapment of the microbial cells in natural gels. In general, immobilized systems show higher acid producing and rock phosphate solubilizing activity than freely suspended cells. The potential of gel-entrapped P-solubilizers and mycorrhizal fungi as microbial soil inoculants is also pointed out. Some advantages and constraints of using immobilized cells are discussed and a special emphasis on further research is given. PMID- 11499581 TI - Phytoremediation of organic contaminants in soils. AB - Soil pollution, a very important environmental problem, has been attracting considerable public attention over the last decades. Unfortunately, the enormous costs associated with the removal of pollutants from soils by means of traditional physicochemical methods have been encouraging companies to ignore the problem. Phytoremediation is an emerging technology that uses plants to clean up pollutants in the environment. As overwhelmingly positive results have become available regarding the ability of plants to degrade certain organic compounds, more and more people are getting involved in the phytoremediation of organic contaminants. Phytoremediation of organics appears a very promising technology for the removal of these contaminants from polluted sites. PMID- 11499582 TI - Production of monomeric phenols by thermochemical conversion of biomass: a review. AB - Biomass is a renewable and alternative source for the production of fuels and chemicals. This paper provides a brief survey of lignin precursors as well as thermogravimetric and pyrolysis studies of lignin with special reference to the production of phenols. Thermogravimetric analysis provides information on pyrolysis kinetics while thermogravimetry in combination with mass or infrared spectrometers allowed a rapid characterization of the vapours produced by thermal treatment. Pyrolysis enabled even greater insight into the thermal behaviour of lignin. Pyrolysis of single, dimeric and trimeric model lignin compounds can determine the thermal stability of the intermediate compounds formed and the origin of the pyrolysis products. A free radical mechanism has been suggested as a major route during the early lignin degradation stages followed by a combined free radical and concerted pathway at elevated temperatures. Pyrolysis of lignin in the presence of catalysts as additives was investigated. Significant differences in terms of yields of pyrolysis products and phenolic compounds were observed. The addition of salts resulted in a high weight loss at low temperature and yielded more char than untreated wood. Some metal catalysts such as transition metals and metal oxides such as Fe2O3 and Cu exhibited a better activity in terms of selectivity for the degradation of lignin. PMID- 11499583 TI - Paravalvular leakage after mitral valve replacement: is left atrial enlargement an additional indication for reoperation? AB - BACKGROUND AND AIM OF THE STUDY: Prostheses used to treat heart valve disease improve patient survival, but have certain disadvantages. Paravalvular leakage (PVL) is a rare complication after mitral valve replacement (MVR), and can impair cardiac function and reduce the patient's functional capacity, depending on the degree of periprosthetic regurgitation. METHODS: Between 1985 and July 1999, 2,502 patients underwent MVR with or without concomitant cardiac procedures. Of these patients, 33 (18 males, 15 females; mean age 39.8+/-15.3 years; range: 12 62 years) had PVL of differing degree. The interval between MVR and observation of PVL was 30.5+/-31.5 months (range: 1-126 months), and the period after diagnosis was 22.6+/-31.5 months (range: 2-114 months). Fourteen patients (42.4%) underwent reoperation (RO group), and 19 (57.6%) were followed medically (ME group). Indications for reoperation were reduction of functional capacity, echocardiographically proven serious mitral regurgitation, and hemolysis. RESULTS: Reoperative mortality was 3.0% (1/33), and late mortality 3.1% (1/32) for all patients. Cumulative survival after PVL was 90.2+/-6.7% at both five and ten years. Annular calcification (33.0%) and infective endocarditis (18.2%) were important predictive factors for development of PVL. Only one patient required second re-do surgery. Univariate and forward stepwise logistic regression analyses showed that there was no predictor for the development of severe PVL requiring a second reoperation. No difference was observed between left ventricular dimensions before and after periprosthetic regurgitation. The only significant finding between groups was an increase in left atrial diameter in RO patients after the development of PVL (p <0.05). CONCLUSION: Among patients undergoing MVR there are no clinical features to distinguish who will develop severe PVL during follow up. If PVL reduces the patient's functional capacity or causes serious hemolysis, or if severe PVL is evaluated echocardiographically, then reoperation must be performed. Mild or moderate mitral regurgitation without impairment of functional capacity may be followed medically. In asymptomatic patients, enlargement (>5%) of the left atrial diameter following development of moderate PVL may be a valuable criterion for deciding when to reoperate. PMID- 11499584 TI - Does the morphology of mitral paravalvular leaks influence symptoms and hemolysis? AB - BACKGROUND AND AIM OF THE STUDY: Prosthetic mitral valve replacement (MVR) is associated with paravalvular leak in up to 12.5% of patients. The influence of the morphology and location of paravalvular leaks on clinical symptoms and degree of leak-related hemolysis is unknown. METHODS: Morphology, size, location and number of paravalvular leaks were analyzed in 96 consecutive patients with primary mitral paravalvular leaks. RESULTS: Mitral leak was diagnosed a median of 119 days after primary MVR. A small (1-2 mm) paravalvular leak was found in 41 patients (43%), an intermediate leak (3-5 mm) in 26 (27%), and a large leak (6-15 mm) in 29 (30%). Single leaks were observed in 70 patients (73%), whilst 26 (27%) had multiple leaks. Paravalvular leaks occurred around the entire prosthetic circumference, but were seen predominantly around the mitral commissural areas (76%). The larger the size of the leak, the more symptomatic the patient (p = 0.006); 80% of patients with small leaks were in NYHA classes I and II, whilst 62% with intermediate/large leaks were in NYHA classes III and IV. The number of leaks was not correlated with severity of clinical symptoms. Multiple leaks were more likely to cause significant hemolysis. Patients with preoperative chronic renal insufficiency, postoperative infection or large (>5 cm) left atria were more likely to develop multiple leaks. The size and location of the leaks was surgeon-dependent. CONCLUSION: Intraoperative transesophageal echocardiography is mandatory to detect possible small leaks and technical errors. Strict monitoring of all MVR patients is necessary for prolonged periods, as the appearance of paravalvular leaks is not necessarily correlated with clinical symptoms. Small paravalvular leaks, in particular, may go unnoticed. As the location and size of the leaks were significantly surgeon-dependent, self-monitoring should be mandatory for all surgeons. PMID- 11499585 TI - Erythropoietin can obviate the need for repeated heart valve replacement in high risk patients with severe mechanical hemolytic anemia: case reports and literature review. AB - BACKGROUND AND AIM OF THE STUDY: Brisk hemolysis due to perivalvular leak is usually an indication for valve re-replacement. Repeated surgery after multiple previous valve operations is associated with high mortality, morbidity and failure rates. The present study evaluated the role of erythropoietin (EPO) administration in deferring or obviating the need for repeated surgery. METHODS: Three patients (two men, one woman; age range 62-76 years) with two mechanical valves each and two to four previous heart valve operations, who suffered from severe mechanical hemolytic anemia, were given subcutaneous EPO for 15-17 months. RESULTS: A marked reduction in red blood cell consumption was achieved with a weekly EPO dose of 18,000 U in two patients, both of whom also had mild or moderate kidney malfunction. A third patient with normal renal function and extreme hemolysis showed a transient, partial response to 30,000 U of EPO per week, and eventually needed a fifth operation. CONCLUSION: EPO may defer or even obviate the need for repeated valve surgery in patients with severe hemolysis due to perivalvular leak, especially those with inadequate EPO response, such as those with renal malfunction. PMID- 11499586 TI - Hemodynamics and early performance of the St. Jude Medical Regent aortic valve prosthesis. AB - BACKGROUND AND AIM OF THE STUDY: The St. Jude Medical (SJM) Regent heart valve is a new bileaflet prosthetic valve modified from the currently marketed SJM mechanical valve, with a modified external profile that results in a larger geometric orifice area without changing the existing design of the pivot mechanism or blood contact surface areas. The aim of the present study was to report the early hemodynamic and clinical results of an on-going multicenter trial investigating the clinical performance of the Regent mechanical aortic valve prosthesis. METHODS: The early results from 204 patients at 11 centers in North America and Europe who underwent implantation of a Regent mechanical aortic valve prosthesis are described. Clinical status was prospectively recorded, and echocardiography with Doppler performed at hospital discharge, and at two and six months and one year postoperatively. RESULTS: Follow up to date is 109.3 patient years (average follow up 0.5+/-0.4 years per patient; range: 0 to 1.7 years). NYHA class improved for the group, and there were low rates of clinical adverse effects. Echocardiographic mean pressure gradient at six months was 13.8+/-10.3, 7.4+/-4.1, 5.4+/-3.2, 5.2+/-2.8, 3.4+/-2.3 and 3.6 mmHg, respectively, for 19, 21, 23, 25, 27 and 29 mm valves; effective orifice area was 1.5+/-0.6, 2.0+/-0.7, 2.4+/-1.0, 2.5+/-0.7, 3.6+/-1.4 and 4.8 cm2, respectively. There was a statistically significant decrease in left ventricular mass index between early postoperative (169.1+/-57.7 g/m2) and six months follow up (137.2+/-42.7 g/m2, delta = -30.1+/-42.5 g/m2, p <0.0001). CONCLUSION: The SJM Regent aortic valve has excellent associated hemodynamics with rapid and significant left ventricular mass regression. In all cases of adverse events, rates fell within Objective Performance Criteria guidelines. Long-term clinical assessment is on-going. PMID- 11499587 TI - Structural durability in Carpentier Edwards Standard bioprosthesis in the mitral position: a 20-year experience. AB - BACKGROUND AND AIM OF THE STUDY: Few long-term data are available on the Carpentier-Edwards Standard bioprosthesis in the mitral position. As for other bioprostheses, patient age at the time of implant is the main risk factor for structural deterioration, but no published report has analyzed the life-span of these bioprostheses with respect to this parameter. METHODS: A series of 139 patients who underwent mitral valve replacement with the Carpentier-Edwards Standard bioprosthesis between 1978 and 1987 was reviewed. Mean age at implant was 59.6+/-14.7 years (range: 17-79 years). Follow up was 98.4% complete; total follow up was 1,078.7 patient-years (pt-yr) (mean 8.4+/-4.1 years). Mean follow up in the subgroup of patients alive at the time of the survey was 10.4+/-3.4 years. RESULTS: Structural valve deterioration (SVD) occurred in 30 patients, with mean time to onset of deterioration 9.0+/-2.7 years (median 8.7 years). This time was independent of age at the time of implantation. Analysis by age group (< or =35, 36-50, 51-60, 61-65, 66-70, >70 years) showed deterioration to be more frequent in younger subjects (linear rates 7.9, 6.0, 3.3, 2.4, 0.6 and 0.4% pt yr, respectively). Over the age of 65 years, the risk of SVD no longer varied with age, and was a rare complication. CONCLUSION: The mean time to onset of SVD was independent of patient age at the time of implant. After 65 years, the risk of SVD was low, without any significant variation. The Carpentier-Edwards Standard bioprosthesis may be used in the mitral position in subjects aged over 65 years, and with a low risk of deterioration. PMID- 11499588 TI - Invited comment: mean time to 'actual' failure for porcine heart valves. PMID- 11499589 TI - Aortic valve reoperation after homograft or autograft replacement. AB - BACKGROUND AND AIM OF THE STUDY: With increasing use of homograft and autograft aortic valves for aortic valve replacement (AVR), more patients will be presenting for aortic valve reoperation due to structural degeneration of the homograft or autograft valve. Management options include homograft re replacement, which may require extensive surgery, versus AVR with a mechanical valve or a stented xenograft. Here, results are reported in 18 consecutive patients who underwent aortic valve re-replacement (AVreR) after previous homograft or autograft insertion. METHODS: Between May 1976 and March 2001, 18 patients underwent AVR after previous homograft (n = 16) or autograft (n = 2) insertion. The homograft or autograft had been implanted as a full root in eight patients (44%), as a mini-root in one (6%), and in the subcoronary position in nine (50%). Indication for the reoperation was structural valve degeneration (n = 14; 72%) in one occasion combined with aneurysm of the homograft, or endocarditis (n = 4; 22%), and seven (39%) presented as a non-elective procedure. The median interval between the two operations was 5.4 years (range: 0.3-10.8 years). RESULTS: Fourteen patients (78%) received either a mechanical valve (n = 12; 67%) or a stented xenograft valve (n = 2; 11%). Four others (22%) required root re replacement with either another homograft (n = 3) or a mechanical valved conduit (n = 1) for endocarditis (n = 2) or an associated aneurysm (n = 2). Overall hospital mortality was 11% (n = 2) due to stroke (n = 1) or respiratory failure (n = 1). Two patients died 3.1 and 7.0 years after the procedure. CONCLUSION: Aortic valve reoperation after previous homograft or autograft implantation is a rare operation and presents a high-risk group. A simplified approach was preferred by utilizing mechanical or stented xenograft valves at reoperation, while homograft re-replacement was reserved for endocarditis or an associated aneurysm. PMID- 11499590 TI - Editorial: Are immune mechanisms important in tissue heart valve failure? A debate. PMID- 11499591 TI - The possible role of immune responses in bioprosthetic heart valve failure. PMID- 11499592 TI - Don't blame the lymphocyte: immunologic processes are NOT important in tissue valve failure. PMID- 11499593 TI - Tissue damage and calcification may be independent mechanisms of bioprosthetic heart valve failure. AB - BACKGROUND AND AIM OF THE STUDY: Porcine bioprosthetic valves have excellent hemodynamics and do not require anticoagulation, but have limited durability. Cusp tearing is a major cause of bioprosthetic valve failure. It has been suggested that the mechanism of bioprosthetic valve failure is stiffening by calcification, which leads to elevated stresses and secondary collagen fiber damage and leaflet tearing. This thesis was tested in explanted porcine bioprostheses. METHODS: A total of 60 explanted porcine bioprosthetic valves was tested mechanically, and 15 explanted valves were examined grossly and histologically. Circumferentially and radially oriented samples of cusp tissue were tested uniaxially in a materials testing machine and compared with five controls. RESULTS: Mean (+/-SD) duration of implantation was 10.9+/-5.6 years. Circumferential specimens from explants were less extensible than controls (11.0+/-5.5% versus 24.5+/-2.8% strain, p <0.001), and failed at lower tensions (973+/-733 versus 3075+/-911 N/m, p = 0.001) and at lower strains (21.2+/-8.1% versus 47.3+/-7.1% strain, p <0.001). Radial specimens from explants were less extensible (28.7+/-6.8% versus 39.2+/-5.9% strain, p = 0.002) and failed at lower strains (60.3+/-17.3% versus 112.2+/-24.9% strain, p <0.001) than the controls. The stiffness of the explants was unchanged from controls in both circumferential and radial samples. There were no differences between explants and controls in radial and circumferential stiffness, and in radial failure strength. Calcification was mild and diffuse in most of the tested samples. Tears were found in areas without calcific deposits, along with breaks in collagen fiber bundles. CONCLUSION: These results do not support the thesis that calcification stiffens glutaraldehyde-fixed porcine bioprostheses, except when the entire cusp is transformed into a solid mass of mineral. Rather, leaflet tears may develop as a result of accumulated mechanical damage that is independent of calcification. PMID- 11499594 TI - Infective endocarditis of native cardiac valves: 22 years' surgical experience. AB - BACKGROUND AND AIM OF THE STUDY: Most previous studies on surgery for endocarditis included cases of both prosthetic and native valve endocarditis (NVE), which clearly differ in their course and prognosis. METHODS: Between February 1979 and July 2000, 199 patients underwent surgical treatment for NVE. The native valve was replaced in 185 patients, and repaired in 14. Correlations between previous heart disease and site of NVE, site and etiology, site and embolism, and etiology and extracardiac infection were analyzed. Viability of echocardiography to identify NVE features was compared with surgical inspection. Risk factors for hospital, late mortality and recurrence were assessed by multivariate analysis, including preoperative, operative and postoperative variables. RESULTS: Streptococci were found in 52 cases (26.1%), mostly spreading from splanchnic (n = 10) and cutaneous (n = 9) infections; Staphylococci were found in 43 cases (21.6%), mostly from cutaneous foci (n = 20). Tricuspid valve endocarditis was due to Staphylococcus sp. in all documented cases (11/15; p <0.001), while Streptococcus sp. predominated in the mitral (p = 0.026) position. Echocardiography was reliable in identifying anatomic features of NVE, except for periannular abscess (13 versus 31 found at surgery; p <0.001). Hospital mortality was 6% (n = 12): preoperative NYHA class IV (p = 0.034) and emergency surgery (p = 0.001) were significant predictors of mortality. Average follow up was 57.2+/ 4.4 months and was 96.3% complete. There were 27 late deaths (15%). Actuarial survival was 75.7% at 10 years and 60.3% at 20 years. Low social status (p = 0.022), diabetes mellitus (p = 0.003), fever at time of surgery (p = 0.041) and persistent postoperative fever (p = 0.0002) were predictors for late mortality. Endocarditis recurred in 28 patients (15.5%), with 74.1% freedom at 10 years and 68.7% at 20 years. Drug abuse (p = 0.001), mitral endocarditis (p = 0.002), positive blood cultures (p = 0.0003), homograft implantation (p = 0.0012) and persistent postoperative fever (p <0.0001) were significant predictors for recurrence. CONCLUSION: Preoperative conditions of patients with NVE significantly affect early and long-term outcomes. Prognosis is worsened by delayed diagnosis and operation during the active septic process. PMID- 11499595 TI - Brucella endocarditis: case report and review of the literature. AB - Cardiac involvement in childhood brucellosis is rare and, when present, findings are usually seen in acute rheumatic fever with endocarditis. We report a successfully treated case of Brucella endocarditis in which the aortic valve was affected. Medical therapy did not cure this patient who, due to hemodynamic deterioration, required valve replacement. This case report suggests that successful management of Brucella endocarditis requires a combination of medical and early surgical therapy. PMID- 11499596 TI - Antimineralization treatments in stentless porcine bioprostheses: an experimental study. AB - BACKGROUND AND AIM OF THE STUDY: Photo-oxidation treatment of porcine stentless bioprostheses (Photofix) was compared with glutaraldehyde fixation, with either AOA (Freestyle valve) or Tween-80 (Edwards Prima Plus valve). METHODS: Six valves of each type were implanted in juvenile sheep, in the pulmonary position. Valves were explanted after three or six months and examined macroscopically, by X radiography, and by light and transmission electron microscopy. Calcium content was determined by atomic absorption spectrometry. RESULTS: The cusps of all valves were free of calcification, and had normal histology and function. Calcium contents (median +/- IQR) were 0.63+/-0.45, 0.73+/-1.46 and 0.46+/-1.42 microg/mg for the Photofix, Freestyle and Prima Plus valves, respectively (p = NS). Calcium contents of the aortic wall portions were 0.71+/-1.27 (Photofix), 10.78+/-77.22 (Freestyle) and 28.70+/-66.53 (Prima Plus) (p <0.05 for Photofix versus Freestyle or Prima Plus). CONCLUSION: Photo-oxidation of a porcine stentless valve prevents calcification not only in the cusps, but also in the aortic wall portion. PMID- 11499597 TI - A general method for estimating deformation and forces imposed in vivo on bioprosthetic heart valves with flexible annuli: in vitro and animal validation studies. AB - BACKGROUND AND AIM OF THE STUDY: The use of flexible structures within cardiovascular prostheses such as valves, stents and vascular grafts has been proposed as a means of more closely modeling native mechanics, and thereby reducing the biomechanical problems associated with rigid materials. However, the design of such materials has been hampered by the paucity of quantitative information on the in-vivo behavior of such structures. The aim of this study was to explore the use of 3D ultrasound imaging coupled with finite element analysis (FEA) as a tool to estimate deformation and forces imposed in vivo on a novel bioprosthetic valve design. METHODS: The method was first tested using in-vitro static loading conditions, where good agreement between displacements seen on video and those obtained from application of the identical force within the finite element program was seen. The method was then tested in a porcine model with valves implanted in the mitral position. Images of the deforming annular ring were obtained over the cardiac cycle using 3D intravascular ultrasound; these images were fed into the FEA program for calculation of reaction forces. RESULTS: Results in vitro showed that a force of 2.7-8.0 Newtons (N) was required to produce a deformation of between 1.0 and 3.0 mm in the radial direction. A time history of deformation and force around the ring of the valve stent could be obtained for the in-vivo conditions. These results revealed a maximum deformation of 0.5-1.7 mm along the short axis (anteroposterior) of the mitral valve. Coupled to this, a peak reaction force of 4.4-13.9 N was found at the points corresponding to maximal deflection. Both deformation and reaction force reached maximum during atrial contraction. CONCLUSION: This method provides an accurate means of estimating deformation and corresponding forces imposed in vivo on intracardiac prostheses. The results provide information on the dynamic behavior of the mitral valve annulus. Such information should be useful in the design of flexible cardiovascular prostheses. PMID- 11499598 TI - A juvenile sheep model for the long-term evaluation of stentless bioprostheses implanted as aortic root replacements. AB - BACKGROUND AND AIM OF THE STUDY: Orthotopic valve replacement in large animals is an important component of the preclinical assessment of bioprosthetic valves. To provide the most useful preclinical information, the development of models that parallel clinical practice patterns is essential. Therefore, we sought to develop a technically feasible and reproducible model for chronic evaluation of stentless bioprosthetic aortic valves implanted as aortic root replacements in juvenile sheep. METHODS: Juvenile domestic sheep (mean age 21+/-2.28 weeks; range: 17-26 weeks) underwent aortic root replacement using standard cardiopulmonary bypass (CPB) and surgical techniques. Animals were implanted with 19 mm (n = 21), 21 mm (n = 18) or 23 mm (n = 4) bioprostheses from two different manufacturers, and followed for 150 days. Animals surviving at least 150 days were considered long term survivors; those which died prior to postoperative day (POD) 31 were considered operative deaths. RESULTS: Forty-three animals underwent aortic root replacement. The mean CPB time was 91+/-20 min (range: 62-149 min); mean cross clamp time was 63+/-13 min (range: 39-95 min). Thirty-five animals (81%) survived the first 30 days of the study period. Five deaths occurred at POD 0 due to anastomotic complications. One death occurred each on POD 3, 6, and 26 as a result of prosthesis size mismatching, thromboembolic complications, and endocarditis, respectively. There were five late deaths. Twenty animals survived the minimum 150-day study period, and 12 were sacrificed at 183+/-17 days. Six animals remain alive at 151+/-0.98 days, and one animal died each on POD 184 and 190. The remaining 10 animals are not yet 150 days from their operation. Currently, all are well at 102+/-34 days (range: 33-140) days. CONCLUSION: These data suggest that long-term evaluation of stentless aortic bioprostheses implanted as aortic root replacements can be accomplished using juvenile sheep. PMID- 11499599 TI - Is the tricuspid position suitable for testing replacement bioprosthetic valves in the sheep model? AB - BACKGROUND AND AIM OF THE STUDY: Glutaraldehyde may promote calcification in xenograft tissue by the action of toxic aldehyde group residues involved in the cross-link process. Post-fixation treatment with homocysteic acid (HA) neutralizes this toxicity by bonding aldehyde groups, and enhances biocompatibility on the basis of strongly electronegative sulfonic groups. Previous studies in a rat subcutaneous model showed significant long-term mitigation of mineralization of glutaraldehyde-fixed pericardium treated with HA. This study aimed to assess the anticalcific efficacy of HA in a valvular implant in growing sheep, and establish if the tricuspid position is suitable for testing replacement bioprosthetic valves. METHODS: Eleven stented 25 mm Pericarbon bioprostheses (seven HA-treated, four standard) were implanted in the tricuspid position of growing sheep. Infective endocarditis occurred in four prostheses. Among the remaining seven, three (two HA-treated, one standard) were explanted at 91 days (mid-term), and four (two HA-treated, two standard) at 140-141 days (long term). All explants were studied by gross, X-ray, light, transmission and scanning electron microscopy, as well as by atomic absorption spectroscopy. RESULTS: No histological and ultrastructural difference in tissue preservation were observed between HA-treated and standard Pericarbon bioprostheses, either in the mid or long term. The mean calcium content of mid-term HA-treated explants was 9.55 mg/g compared with 16.26 mg/g in mid-term standard explants. Only one late standard explant failed as a result of severe stenosis caused by massive dystrophic calcification. Among four late explants, two showed significant increase in mineralization (HA-treated, 87.45 mg/g; standard, 181.20 mg/g), while two showed calcium contents similar to those in mid-term explants (HA-treated, 11.96 mg/g; standard, 17.32 mg/g). CONCLUSION: Post-fixation treatment with HA preserves structural properties after tricuspid implantation in growing sheep. The tricuspid implant in the sheep model failed to reproduce remarkable accelerated progressive calcification in all xenografts so as to demonstrate a significant difference between HA and standard explants. The tricuspid position for testing replacement bioprosthetic valves should be abandoned, and investigations repeated with the prosthesis in the mitral position. PMID- 11499600 TI - De Vega annuloplasty and Carpentier-Edwards ring annuloplasty for secondary tricuspid regurgitation. AB - BACKGROUND AND AIM OF THE STUDY: Residual or recurrent tricuspid regurgitation (TR) has been reported after several types of surgical repair. The development of late TR is an important complication of left heart surgery. The results of De Vega annuloplasty were compared with those obtained after Carpentier-Edwards ring (CE ring) annuloplasty in patients with secondary TR. METHODS: The records of 45 patients who underwent surgery for secondary TR between January 1995 and July 2000 were reviewed retrospectively. Twenty-eight patients underwent De Vega annuloplasty, and 17 had a CE ring annuloplasty. The groups were similar with respect to associated cardiac lesions. No significant preoperative differences were observed in NYHA functional class, TR grade, and pulmonary artery pressure between the two groups. RESULTS: One CE patient died of left ventricular dysfunction after postoperative bleeding. The 28 De Vega patients and remaining 16 CE patients had an uneventful recovery, and were discharged. Tricuspid ring size after repair was similar between groups. Mean (+/- SD) follow up in the entire patient cohort was 39+/-23 months (range: 6 to 75 months). TR recurrence was rated as grade II or III in 13 patients (45%) after De Vega annuloplasty, but was grade II or III in only one patient (6%) patient after CE ring annuloplasty. There was a significant difference in TR recurrence between the groups (p = 0.027), but no significant difference in NYHA class. CONCLUSION: CE ring annuloplasty significantly decreased the recurrence of TR; thus, CE ring annuloplasty is superior to De Vega annuloplasty in patients with secondary TR. PMID- 11499601 TI - Results of surgical repair of ostium primum atrial septal defect in adult patients. AB - BACKGROUND AND AIM OF THE STUDY: Ostium primum atrial septal defect (ASD I) is very rarely observed in the adults. Although ASD I was first corrected surgically almost 50 years ago, the efficacy of surgical treatment in adults has not been well documented. Thus, the long-term outcome of patients aged over 20 years and having surgical repair of ASD I was examined. METHODS: Fifteen patients (10 males, five females; mean age 31.4+/-13.1 years; range: 20 to 56 years) who had surgical repair of ASD I between 1982 and 2000 were followed. All patients were examined physically and underwent chest radiography, ECG and echocardiography (cross-sectional and Doppler) before and after surgery. Autologous pericardium was used to close the defect in 14 patients, and a direct suture was used in one patient. Mitral valvuloplasty (repair of the cleft) was performed in 12 patients, and De Vega annuloplasty in eight. RESULTS: There were no hospital deaths. In one case a pacemaker was implanted five days after surgery because of complete heart block. Preoperatively, nine patients (60%) were in NYHA classes III and IV; at the end of follow up, 14 (93.3%) were in classes I and II. Preoperatively, 13 patients had sinus rhythm, and one each had atrial fibrillation (AF) and rhythm from the atrioventricular sinus. During follow up, three patients developed AF which was treated successfully with electrical cardioversion. The preoperative mean cardiac volume index of 695 +/- 216 ml/m(2) was reduced significantly after repair to 523 +/- 108 ml/m(2) (p < 0.05). Before surgery, mitral regurgitation was observed (severe in five cases, moderate in seven, mild in three). Postoperatively, a residual intracardiac shunt was identified in one case. Postoperative mitral regurgitation was noted in six patients (moderate in two, mild in four). The right ventricular dimension was decreased significantly, from 5.0 +/- 1.5 mm before surgery to 3.2 +/- 0.6 mm after repair (p < 0.001). CONCLUSION: Patients aged over 20 years with ASD I benefit from surgical repair of the defect. PMID- 11499602 TI - The predictive value of chronic atrial fibrillation for the short- and long-term outcome after percutaneous mitral balloon valvotomy. AB - BACKGROUND AND AIM OF THE STUDY: The predictive value of chronic atrial fibrillation (AF) before percutaneous mitral balloon valvotomy (PMBV) is still under discussion. The effect of the duration of chronic AF on short- or long-term results is not known. Therefore, we analyzed the predictive value of pre procedural chronic AF and the duration of this rhythm disturbance for short- and long-term outcome after PMBV in patients with mitral valve stenosis. METHODS: A total of 140 PMBV procedures was performed in 137 patients with severe mitral stenosis. Sixty-three patients (45%) were in chronic AF; in 40 patients (63%) the AF was of more than one year duration. A successful procedure is defined as PMBV achieved without acute mitral valve replacement, and a mitral valve area after PMBV of > or =1.5 cm2. RESULTS: Patients in chronic AF were significantly older, had a larger left atrial diameter and higher NYHA functional class, compared with patients in sinus rhythm (SR). The success rates of PMBV were 80.5% and 77.6% in patients with SR and AF, respectively (p = NS). Mean follow up was 4.2+/-2.6 years (n = 127). At four years' follow up the event-free survival was 86.5% in patients with SR, and 78.5% in those with chronic AF at baseline (p = 0.031). Multivariate analysis of the entire study population showed the presence of chronic AF to be the only pre-procedural independent predictor for severe mitral regurgitation after PMBV (p = 0.030), as well for an event (p = 0.039) and restenosis (p = 0.034) during follow up. The risk for an event or restenosis during follow up increased seven-fold when chronic AF at baseline was present for more than one year (p = 0.010). CONCLUSION: Pre-procedural chronic AF is an independent predictor for unfavorable outcome at short- and long-term follow up after PMBV. A longer duration of AF further increases the risk of an event or restenosis. PMID- 11499603 TI - Aortic valve repair of congenital bicuspid aortic valve associated with aneurysm of the ascending aorta. AB - Two unusual cases of congenital bicuspid aortic valve associated with aneurysm of the ascending aorta are reported. One patient with a 7-cm ascending aortic dilatation and aortic regurgitation (AR) (II/IV), and another with a 6-cm ascending aorta and AR (III/IV), presented for treatment. Replacement of the ascending aorta and aortic valve repair were performed in both cases. Aortic valve repair included resection of the raphe, leaflet plication and subcommissural annuloplasty. Both patients had satisfactory results in the early postoperative period. Despite the promising outcomes after surgery in these patients, long-term changes in valve function and durability remain unknown. Additional close observation and monitoring are required before the procedure can be recommended as the standard of care. PMID- 11499604 TI - Right atrial thrombus associated with combined valvular disease: case report. AB - A 77-year-old man had a large right atrial (RA) thrombus associated with a combined mitral and tricuspid valve disease. Echocardiography showed a large, immobile, non-homogeneous, irregularly surfaced mass in the dilated RA, and prolapse of the anterior mitral leaflet resulting in massive mitral regurgitation. Computed tomography (CT) revealed a laminated structure with calcification and distinct margins, without invasion to the wall of the RA. On the basis of these echocardiographic and CT findings, a diagnosis of combined mitral and tricuspid valvular disease complicated with RA thrombus was made. Removal of the RA thrombus, mitral valve replacement and tricuspid annuloplasty were performed simultaneously, with successful outcome. PMID- 11499605 TI - Traumatic tricuspid valve insufficiency: case report. AB - A 30-year old man underwent a head-on motor accident and was admitted to hospital, where he was successfully treated for several injuries. He was discharged home with slight tricuspid regurgitation, without signs of hemodynamic decompensation, and required no further treatment. Five years later he was admitted to the National Institute of Cardiology due to overt right ventricular failure secondary to tricuspid valve regurgitation. After several unsuccessful attempts at papillary muscle and chordal repair, a 33-mm SJM Biocor valve was implanted. This case history underlines the value of echocardiography, even after blunt chest trauma without cardiac symptoms. A good functional result can be achieved years after onset of traumatic valve insufficiency, but early diagnosis and surgical treatment increases the feasibility of tricuspid valve repair rather than replacement, and also prevents progressive deterioration of right ventricular function. PMID- 11499606 TI - Thirty years survival without anticoagulation after aortic valve replacement with a Bjork-Shiley prosthesis. AB - Anticoagulation is mandatory in all mechanical valve replacements and, when well managed, permits both embolism and valve thrombosis to be kept at low levels. Here, we report the case of a male patient who has survived 30 years without anticoagulation following aortic valve replacement with the Bjork-Shiley prosthesis. PMID- 11499607 TI - Environmental radioactivity research--expectations in the new millennium. PMID- 11499608 TI - Distribution of U-Th nuclides in the riverine and coastal environments of the tropical southwest coast of India. AB - A reconnaissance study has been made on the distribution of 238U, 234U, 232Th and 230Th in soils, water, suspended particulate matter (SPM) and bottom sediments in the Kali river basin around Kaiga, its estuarine region and the adjacent Arabian Sea to obtain the baseline data of U-Th series nuclides in view of the commissioning of nuclear power reactors at Kaiga, near Karwar, on the southwest coast of India. Drainage basin soils developed over greywackes (the dominant litho-unit upstream) are lower in 238U/Al and 232Th/Al ratios by factors of 3-5 in comparison with those developed over tonalitic gneisses (the dominant litho unit downstream). The dominance of the former type of soils is reflected in the composition of river-bottom sediments derived from the upstream drainage basin during the monsoon. The 232Th in bottom sediments tends to increase towards the estuarine and coastal areas, presumably due to deposition of heavy minerals and onshore transport of coastal sediments into the estuary. The dissolved U in the Kali river is low (0.001-0.02 microg/l) when compared to the major Indian rivers as the Kali river flows through U-poor greywackes. Thus, the input of dissolved U to the Kali estuary is dominated by sea water. Although there is some evidence for the removal of dissolved U at low salinity during estuarine mixing, its behaviour is conservative in the lower estuary (at higher salinities). The removal rate of dissolved U from the Kali river basin is similar to that reported from other tropical river basins. The U flux from all the west-flowing rivers of Peninsular India is estimated at 26.3 x 10(6) g/yr to the Arabian Sea which is about 2% of the flux from the Himalayan rivers to the Bay of Bengal. PMID- 11499609 TI - Comparison of laboratory uranium sorption data with 'in situ distribution coefficients' at the Koongarra uranium deposit, Northern Australia. AB - Distribution coefficients derived from laboratory sorption experiments are commonly used to model the migration of long-lived radionuclides in the environment. However, it has been suggested that field measurements in natural systems ('in situ distribution coefficients') may provide a more accurate indication of 'true' partitioning coefficients than laboratory experiments. In this paper, the relationship between field and laboratory sorption data for uranium is evaluated, using data from the Koongarra uranium deposit in Northern Australia. An extensive suite of laboratory sorption measurements and in situ partitioning data for U has been obtained at this site. A valid comparison can only be made when the calculation of field partitioning is based on U in 'accessible' phases (rather than total U in the solid) and U species in true solution (i.e. excluding particles). In this study, accessible U was estimated using a chemical extraction and the results were verified using an isotope exchange technique. A satisfactory correspondence between field and laboratory partitioning data was obtained when the pH values and partial pressures of CO2 in laboratory sorption experiments were similar to those found in the field. Under these conditions, the measured laboratory sorption ratios (Rd) and in-field partitioning values (Pacc) for U at Koongarra were in the range between approximately 1 x 10(3) and 2 x 10(4) ml/g. However, the distribution of U in solid and groundwater phases at Koongarra is extremely heterogeneous. This variability must be taken into account when modelling radionuclide migration at this site. PMID- 11499610 TI - Measurements of summer radon and its progeny concentrations along with environmental gamma dose rates in Taiwan. AB - The concentrations of 222Rn (radon) and its progeny with surrounding environmental gamma-dose rates were measured simultaneously inside and outside of buildings at 10 locations around Taipei and Hualien in Taiwan. For summer radon in Taiwan, indoor concentrations were estimated to be about 20 Bq m(-3) with about 90 nSv h- of environmental gamma, and outdoors, about 10 Bq m(-3) with about 70 nSv h(-1). The equilibrium factors were calculated to be 0.2-0.3 indoors and 0.3-0.4 outdoors. Indoor radon concentration had a weak positive correlation with gamma-dose rate. Since there is a possibility that high radon concentrations exist indoors during the cool season in Taiwan because of extremely low ventilation rates in the dwellings, a winter survey in January through February will be needed for future estimation of the annual effective dose. PMID- 11499611 TI - Assessment of the global fallout of plutonium isotopes and americium-241 in the soil of the central region of Saudi Arabia. AB - A radiochemical technique for determination of plutonium isotopes and 241Am in soil samples is tested against IAEA-standard reference materials to determine its accuracy and precision for reliable results. The technique is then used in the investigation of topsoil samples, collected from the natural environment of the central region of Saudi Arabia, to assess the effect of fallout accumulation of these radionuclides in the region. Plutonium and americium were sequentially separated from all other components of the sample by anion-exchange chromatography and co-precipitated with Nd3+ as fluorides. The precipitates were mounted on membrane filters and measured using a high-resolution alpha spectrometer. The results of the analysis of the reference materials showed satisfactory sensitivity and precision of the technique. The results of the analyzed soil samples show activity levels ranging from < LLD to 0.089 and from 0.9999) over a concentration range of 3-1,000 ng/ml of rutin in plasma. The lower limit of quantification was ca. 5 ng/ml of rutin in plasma. The detection limit (defined as signal-to-noise ratio of about 3) was approximately 0.75 ng/ml. A preliminary experiment to investigate the plasma concentration of rutin after oral administration of 500 mg of rutin to a healthy volunteer demonstrated that the present method was suitable for determining rutin in human plasma. PMID- 11499621 TI - Simultaneous gas chromatographic determination of concentration and isotopic enrichment of fatty acids in human plasma using flame ionization and mass spectrometric detection. AB - Free fatty acids (FFAs) are important not only because they provide substrate for oxidation but also because they have the potential to regulate several metabolic and hormonal processes. Using stable isotope tracers, these processes can be studied. Here we present a gas chromatographic method to measure FFA concentrations and enrichments after extraction from plasma and subsequent derivatization in one analytical run, using both flame ionization and mass selective detection. For concentration determinations intra-assay variation ranged from 1.5 to 4.9%, inter-assay variation ranged from 3 to 11%. Intra- and inter-assay variations of the enrichment determination of palmitic acid were 1.4 and 0.9%, respectively. PMID- 11499622 TI - Simultaneous supercritical fluid extraction and chemical derivatization for the gas chromatographic-isotope dilution mass spectrometric determination of amphetamine and methamphetamine in urine. AB - An in-situ supercritical fluid extraction (SFE) and chemical derivatization (ChD) procedure followed by gas chromatography-isotope dilution mass spectrometry (GC MS) for the determination of amphetamines in urine is described and evaluated. While using celite as the SFE wet-support, the one-pot sample pretreatment procedure also employs ammonium water to alkalize the urine matrix that contains protonated amphetamine (AP) and methamphetamine (MA). The mean recoveries achieved by simultaneous SFE-ChD, i.e., 95% (RSD=3.8%) for AP and 89% (RSD=4.0%) for MA, are significantly better than the corresponding overall recoveries obtained upon stepwise SFE-ChD, suggesting the unreacted trifluoroacetic anhydride (TFA) in the former procedure has strengthened the extracting power of CO, fluid as has been evidenced by a control test. As to GC-MS analysis, the optimal qualitative ions and quantitative ions of the respective analytes were determined via a rigorous evaluation process. Thus, the regression calibration curves for AP and MA in urine are linear within 100 approximately 50,000 ng/ml, with correlation coefficients typically exceeding 0.999. The limits of detection determined by two methods for AP and MA vary from 19 to 50 ng/ml, and limits of quantitation from 21 to 100 ng/ml. Precisions calculated for the triplicate analyses of AP and MA in a 500-ng/ml spiked control, two real-case samples and two quasi real-case samples, respectively, using regression calibration are typically below 10%. The method is simple and reliable. It may serve as an alternative to the existing confirmatory protocol for forensic urine drug testing. PMID- 11499623 TI - High-performance liquid chromatographic method for the estimation of the novel investigational anti-cancer agent SR271425 and its metabolites in mouse plasma. AB - A simple and reliable HPLC method was developed for the estimation of a new anti cancer agent that belongs to the thioxanthone class, SR271425 in mouse plasma. SR271425, it's metabolites and internal standard (SR233377) were separated from plasma by liquid-liquid extraction using dichloromethane after quenching the plasma proteins with acetonitrile. Chromatography was performed on a reversed phase C18 column using methanol-10 mM phosphate buffer, pH 3.5 (45:55) as mobile phase at a flow-rate of 0.8 ml/min for first 10 min and 1.4 ml/min for the next 15 min with UV-Vis detection at 264 nm and SR233377 as internal standard. The retention times of SR271425 and internal standard were 18.6 and 14.8 min, respectively. The limit of detection was 40 ng/ml and the limit of quantification was 78 ng/ml. This method was also able to detect the three metabolites of SR271425. The intra- and inter-day relative standard deviations were less than 13% at all concentrations. This analytical method was precise and reproducible for pharmacokinetics and metabolism studies of the drug in mice. SR271425 is proceeding to phase I clinical trials in 2001. PMID- 11499624 TI - Determination of 2-hydroxyflutamide in human plasma by high-performance liquid chromatography and its application to pharmacokinetic studies. AB - Flutamide is a potent antiandrogen used for the treatment of prostatic cancer. Flutamide undergoes extensive first-pass metabolism to the pharmacologically active metabolite 2-hydroxyflutamide. A simple, sensitive, precise, accurate and specific HPLC method, using carbamazepine as the internal standard, for the determination of 2-hydroxyflutamide in human plasma was developed and validated. After addition of the internal standard, the analytes were isolated from human plasma by liquid-liquid extraction. The method was linear in the 25 to 1,000 ng/ml concentration range (r>0.999). Recovery for 2-hydroxyflutamide was greater than 91.4% and for internal standard was 93.6%. The limit of quantitation was 25 ng/ml. Inter-batch precision, expressed as the relative standard deviation (RSD), ranged from 4.3 to 7.9%, and accuracy was better than 93.9%. Analysis of 2 hydroxyflutamide concentrations in plasma samples from 16 healthy volunteers following oral administration of 250 mg of flutamide provided the following pharmacokinetic data (mean+/-SD): Cmax, 776 +/- 400 ng/ml; AUC(0-infinity), 5,368 +/- 2,689 ng h/ml; AUC(0-t) 5,005 +/- 2,605 ng h/ml; Tmax 2.6 +/- 1.6 h; elimination half-life, 5.2 +/- 2.0 h. PMID- 11499625 TI - High-performance liquid chromatographic method with electrochemical detection for the analysis of O6-methylguanine. AB - An improved system consisting of a combination of high-performance liquid chromatographic methods with electrochemical detection for the separation and analysis of the DNA adduct O6-methylguanine (O6MG) has been developed. This adduct is produced by the interaction of methylating agents with DNA and induces mispairing in the DNA of the target cells. A good separation of modified from unmodified bases is first achieved with an HPLC system using a Partisil 10 SCX column and a salt gradient. A second HPLC step with electrochemical detection and a C18 column is used for farther separation and quantitation of O6-methylguanine. This method shows a linear response up to 15 pg of 06MG tested. The lowest amount detected was 0.5 pg of O6MG and is highly reproducible. This method is useful to study DNA damage as a product of cellular metabolism and its effects on the process of carcinogenesis. PMID- 11499626 TI - Use of molecularly imprinted polymers in the solid-phase extraction of clenbuterol from animal feeds and biological matrices. AB - Clenbuterol molecularly imprinted polymers (MIPs) as chromatographic stationary phase for the solid-phase extraction (SPE) of the drug from biological samples have been prepared. Propylene columns filled with 500 mg of clenbuterol MIPs have been tested with respect to their loading capacity, memory effects, selectivity toward related drugs (mabuterol, clenproperol, clenisopenterol, ritodrine) and specificity toward interferences arising from heterogeneous matrices such as animal feeds, bovine urine and liver. Analytes were concentrated on Extrelut 20 columns and the residues resuspended in 70% acetonitrile. Application, washing and elution fractions were collected and analyzed by HPLC-diode array detection. Results indicate this MIP approach in SPE is extremely selective for clenbuterol, mabuterol, clenproperol and clenisopenterol (>95% found in the eluate), with a loading capacity of about 20 microg/100 mg of stationary phase. Ritodrine showed a recovery rate of 51%. The molecular recognition mechanism is so specific to allow clenbuterol detection and identification by conventional detectors at level of interest (ppb) also from complex matrices such as feeds, urine and liver. PMID- 11499627 TI - Detection of the isoflavone aglycones genistein and daidzein in urine using solid phase microextraction-high-performance liquid chromatography-electrospray ionization mass spectrometry. AB - An improved method of detection of the isoflavone aglycones, genistein and daidzein, is reported using solid-phase microextraction-high-performance liquid chromatography-electrospray ionization mass spectrometry (SPME-HPLC-ESI-MS). Extraction of the isoflavonoids from urine using SPME with a Carbowax-templated resin fiber coating allows rapid preconcentration of the analytes without the usual sample preparation required by other methods. Detection of the analytes is accomplished by HPLC-ESI-MS. Analysis of spiked samples of urine resulted in a linear range of 0.25 to 250 ng/ml for daidzein and 0.27 to 27.0 ng/ml for genistein. Limits of detection of daidzein and genistein were measured at 25.4 pg/ml for daidzein and 2.70 pg/ml for genistein. Daidzein and genistein were detected in urine following consumption of a soy drink. PMID- 11499628 TI - Analysis of 3,5,6-trichloro-2-pyridinol in urine samples from the general population using gas chromatography-mass spectrometry after steam distillation and solid-phase extraction. AB - We have developed a new method for the quantitative trace determination of 3,5,6 trichloro-2-pyridinol (TCPyr). TCPyr is a urinary metabolite specific to the organophosphorus pesticides chlorpyrifos and chlorpyrifos-methyl. After hydrolysis and separation of TCPyr from the urinary matrix using semi-automated steam distillation and solid-phase extraction on a new polystyrol-divinylbenzene copolymer (Isolute 101) the analyte was converted into its tert butyldimethylsilyl derivative by N-tert-butyldimethylsilyl-N methyltrifluoroacetamide (MTBSTFA). Separation and quantitative analysis were carried out by capillary gas chromatography and mass selective detection in selected ion monitoring mode. 2,6-Dibromophenol (DBP) was used as the internal standard. The detection limit was 0.05 microg/l; the limit of quantification was 0.1 microg/l urine. The relative standard deviation of the within-series imprecision was 4.2% at a concentration of 3.5 microg/l. The relative recovery was 104%. The new method was used to analyse the urine samples of 12 persons from the general population without known exposure to the above-mentioned pesticides. TCPyr concentrations between 0.27 and 6.6 microg/l urine were detected in all urine samples. This indicates that there is a baseline excretion of TCPyr in the general population. Four urine samples collected from workers who had applied chlorpyrifos were also analysed. In these samples TCPyr was found in concentrations from 4.7 to 7.9 microg/l. PMID- 11499629 TI - Simultaneous determination of F-beta-alanine and beta-alanine in plasma and urine with dual-column reversed-phase high-performance liquid chromatography. AB - F-beta-Alanine and beta-alanine were detected in plasma and urine samples with fluorescence detection of orthophthaldialdehyde derivatives of F-beta-alanine and beta-alanine after separation with dual-column reversed-phase HPLC. The detection limits of F-beta-alanine and beta-alanine in the HPLC system were approximately 0.3 and 0.7 pmol, respectively. The procedure proved to be very reproducible with intra-assay RSDs and inter-assay RSDs being less than 8%. The usefulness of the method was demonstrated by the analysis of the F-beta-alanine and beta-alanine concentrations in plasma and urine samples from tumor patients treated with S-1 (Tegafur, 5-chloro-2,4-dihydroxypyridine and potassium oxonate in a molar ratio of 1:0.4:1). PMID- 11499630 TI - Simultaneous determination of olanzapine, clozapine and demethylated metabolites in serum by on-line column-switching high-performance liquid chromatography. AB - An automated method for simultaneous routine quantification of the antipsychotic drugs clozapine, olanzapine and their demethylated metabolites is described. The method included adsorption on a cyanopropyl (CPS) coated clean-up column (10 microm; 10 x 2.0 mm I.D.), washing off interfering serum constituents to waste, and separation on C18 ODS Hypersil reversed phase material (5 microm; 250 x 4.6 mm I.D.) using acetonitrile-water-tetramethylethylenediamine (37:62.6:0.4, v/v/v) adjusted to pH 6.5 with concentrated acetic acid. UV-detection was performed at 254 nm. The limit of quantification was 10-20 ng/ml. Relative day to day standard variations ranged between 4.5 and 13.5%. The method is suitable for routine monitoring of olanzapine and clozapine including their demethylated metabolites. PMID- 11499631 TI - Determination of residues of enrofloxacin and its metabolite ciprofloxacin in biological materials by capillary electrophoresis. AB - A method for the analysis of enrofloxacin and ciprofloxacin in chicken muscle using marbofloxacin as internal standard is proposed. Clean-up and pre concentration of the samples are effected by solid-phase extraction and determination is carried out by capillary electrophoresis using a photodiode array detector. The calibration graphs are linear for enrofloxacin and ciprofloxacin from 10 to 300 microg/kg. The method recoveries for enrofloxacin and ciprofloxacin are 74 and 54%, respectively. The limit of detection for the two compounds is lower than 25 microg/kg, which allows the detection of positive muscle samples at the required maximum residue limits. PMID- 11499632 TI - Investigations into the interaction between tumor-inhibiting ruthenium(III) complexes and nucleotides by capillary electrophoresis. AB - Ruthenium(III) complexes of the general formula HL[RuCl4L2], with two trans standing heterocyclic ligands L bound to ruthenium via nitrogen, show remarkable activity in different tumor models. To obtain a deeper insight into the mode of action of this class of anticancer compounds, we investigated the interaction of HIm trans-[RuC14(im)2] (im, imidazole) and HInd trans-[RuCl4(ind)2] (ind, indazole) with all four nucleoside monophosphates in buffered solution by means of capillary electrophoresis. A preference for GMP- and AMP-coordination was found. A decrease of the pH resulted in a significantly increased amount of bound nucleotide. This feature seems to be interesting with regard to the lower pH values in solid tumors. PMID- 11499633 TI - Determination of mephenoxalone in human plasma sample by high-performance liquid chromatography-fluorescence detection. AB - A simple and sensitive high-performance liquid chromatographic method involving fluorescence detection was developed for the determination of mephenoxalone in human plasma. A Cosmosil 5C18-MS column (250 mm x 4.6 mm I.D., 5 microm) was used as stationary phase and the mobile phase consisted of water-acetic acid acetonitrile (200:1:300) at a flow-rate of 1.0 ml/min. The fluorescence absorbance was monitored at 280 nm for excitation wavelength and 310 nm for emission wavelength. Temperature control was kept at 40 degrees C for the column. The limit of quantitation achieved was 10 ng/ml, and the standard curve was found to be linear in the concentration range of 10-10,000 ng/ml. Under these analytical conditions, a sufficient mephenoxalone plasma concentration profile could be obtained for pharmacokinetic study. PMID- 11499634 TI - Quantitative determination of CGS 26214, a cholesterol lowering agent, in human plasma using negative electrospray ionization liquid chromatography-tandem mass spectrometry. AB - CGS 26214 is a synthetic cholesterol-lowering agent shown to be active in the rat, dog and monkey. The present work was conducted to develop a sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for quantitative determination of the compound in human plasma following clinical doses of 10-100 microg per day. A number of analytical challenges were encountered during the development of the assay. The compound was an ester and susceptible to hydrolysis under experimental conditions. A lower limit of quantitation of 50 pg/ml was needed due to the low clinical doses. Positive electrospray ionization of CGS 26214 yielded insufficient sensitivity needed for the studies. Consequently, LC MS-MS conditions were optimized for the negative ion mode of detection. The sample preparation steps proved to be critical in order to reduce the possibility of microbore column (50 mm x 1.0 mm I.D.) obstruction, chromatographic deterioration, and matrix mediated electrospray ion suppression. The present method addressed the above issues. The method was accurate and reproducible and was successfully applied to generate plasma concentration-time profiles for human subjects after low oral doses of the compound. PMID- 11499635 TI - Vasculitis of the nervous system. AB - Vasculitis is inflammation of the blood vessels, which may involve either the central nervous system (CNS), or the peripheral nervous system (PNS), or both. This involvement may be primary and restricted to the CNS, and rarely to the PNS."Primary angiitis of the CNS" is the term used to describe isolated CNS involvement by vasculitis, in which neither the clinical presentation and behaviour of the disease, nor the histopathology is uniform. This heterogeneity indicates a spectrum, depending on the type and extent of the vascular involvement seen within the CNS, covering a group of disorders, rather than a single disease. This may explain the different prognosis and response to treatments. In clinical practice vasculitis of the nervous system, secondary to a known cause or underlying disease is more commonly seen than as a primary disorder. Primary systemic vasculitides and connective-tissue disorders, Behcet's Disease, lymphoproliferative diseases and other malignancies, some infections and related conditions, drugs and substance abuse are some of the conditions known to cause vasculitis in the nervous system. There is a broad variety of pathogenetic mechanisms. Both the CNS and the PNS may be involved, either separately or together. PMID- 11499636 TI - Viral infections of the CNS with special emphasis on herpes simplex infections. AB - Within the past decade the management of acute HSV I encephalitis has been improved dramatically by the advent of the polymerase chain reaction (PCR), a method which has become the gold standard of diagnosis of HSV I encephalitis, replacing diagnostic uncertainties and, avoiding, in particular, invasive brain biopsy. Early detection of HSV II in the neonate is mandatory; however, prevention by Caesarean section and/or prenatal therapy of the mother are for this the best option. Very recently the causative agent of Mollaret's meningitis has proved to be, at least in part, HSV I or II. So far prospective randomized therapeutic trials are awaited for the treatment of Mollaret's meningitis using intravenous acyclovir or the more modern oral forms of virostatics (famciclovir, valaciclovir). For decades the causative agent of facial palsy (Bell's palsy) has been sought; only with the advent of PCR has this question been answered. Although one single study indicates the superiority of a combination of acyclovir plus prednisone, this finding has to be confirmed by a large scale prospective randomised double blind study. Nevertheless, if other causes for the clinical/neurological syndrome of peripheral facial palsy have been excluded, a combination therapy with acyclovir plus prednisone seems to be indicated in a patient with Bell's palsy. PMID- 11499637 TI - Clinical characteristics of the geste antagoniste in cervical dystonia. AB - The geste antagoniste (moving an arm to the face or head) is a well-known clinical feature in cervical dystonia (CD) to alleviate the abnormal posture. The clinical phenomenology of these manoeuvres has not so far been assessed systematically. Fifty patients with idiopathic CD aware of at least one geste antagoniste (60% women, mean age at onset 44.1 years, mean disease duration 7.5 years) were subjected to a standardized investigation including a semiquantitative clinical rating scale and polymyographic recordings of six cervical muscles. Twenty-seven patients (54%) demonstrated more than one geste antagoniste (range 2-5). A clinically significant (> or = 30%) reduction of head deviation was observed in 41 patients (82 %). Dystonic head posture improved by a mean of 60 % along all planes by the geste manoeuvre with a complete cessation of head oscillations in nine of 33 patients (27 %) with phasic CD. No significant laterality of the "geste-arm" or the facial target area was found. The duration of geste-effects depended significantly on disease duration and determined the patient's self-rating of the benefit of the manoeuvre. EMG-polygraphy revealed two types of geste-induced polymyographic changes: a decrease in recruitment density and amplitude in at least one dystonic muscle (66%), and an increased tonic muscle activation in the remaining patients. The remarkable efficacy of the geste antagoniste and the considerable variety in performance, duration, and EMG pattern of these manoeuvres warrant further investigation of the therapeutic use of sensorimotor stimulation, in particular for those CD patients who experience limited or no effect from botulinum toxin therapy. PMID- 11499638 TI - Long-term sensory deficit after Guillain-Barre syndrome. AB - In order to document the sensory deficit still present several years after onset of Guillain-Barre syndrome (GBS) and to determine if the sensory residua have a disrupting effect on daily life, 122 patients were asked to cooperate in a neurological examination and to complete a questionnaire three to six years after onset. On functional assessment 84 patients had no or only minor neurological symptoms or signs, 24 patients showed moderate recovery and 14 patients were left with severe residual signs. On neurological examination, residual sensory deficit was found in the arms of 38 % of the patients and in the legs of 66 % of the patients. Sensory disturbance was experienced as moderate to severe in the arms of 27 % of the patients and in the legs of 40 % of the patients. Muscle aches and cramps were still present in 48 %. There was a statistically significant relation between muscle aches and cramps and objective residual sensory deficit but not with residual weakness. Furthermore, in the group of patients with a pure motor GBS, significantly fewer people suffered from muscle aches and cramps than in the remaining patients (p=0.04). Twenty-five percent of patients changed jobs after their illness, and 44% gave up some leisure activities. It can be concluded that many patients still suffer from sensory deficit, and a considerable number experience these as moderately to seriously disruptive, especially in the legs. Muscle aches and cramps seems to be related to sensory rather than motor dysfunction. PMID- 11499639 TI - Serum inflammatory markers and clinical/MRI markers of disease progression in multiple sclerosis. AB - The aim of this study was to assess whether mean serum levels of inflammatory markers when measured serially correlate with disease progression or putative MRI markers of axonal loss in a cohort of well-characterised multiple sclerosis (MS) patients. Serial serum levels of soluble vascular cell adhesion molecule-1 (sVCAM 1), soluble intercellular adhesion molecule-1 (sICAM-1), nitric oxide metabolites nitrate and nitrite (NOx), C-reactive protein (CRP), neopterin and tumour necrosis factor alpha (TNF-alpha) were measured in 29 MS patients, 13 with a relapsing remitting (RR) and 16 with a secondary progressive (SP) course, who were participating in a phase II clinical trial of anti-CD4 therapy. Short-term whole blood stimulated TNF-alpha production was also measured. Patients were studied 12 times over an 18-month period. MRI variables included the number and volume of Gd-enhancing lesions and the change in T2-hyperintense, T1-hypointense lesions and cerebral volume between the baseline and exit studies. Disease progression required a sustained change in the EDSS. There was no correlation between mean levels of inflammatory markers over the study period and disease progression or changes in any of the MRI measures. However, mean sICAM-1 levels from the first 6 months of the study were higher in patients who progressed during the study than in those that did not (443 (SD439) vs. 235 (SD162) ng/mL, p=0.05). Mean levels of NOx were significantly higher in patients with RR MS than in patients with SP disease (59.1 micromol/L (SD 12.8) vs. 48.7 micromol/L (SD 11.9), p=0.02). Patients with either a single relapse or no relapse had significantly higher NOx levels than to patients with multiple relapses during the 18 month follow-up (59.0 micromol/L (SD 12.3) vs. 47.9 micromol/L (SD 12.0), p=0.02). The mean levels of the other inflammatory markers did not correlate with disease course or relapse-rate. Serum levels of many inflammatory markers do not correlate with short-term disease progression. Some of the data suggest that the effects of inflammation on disease progression are delayed. In addition raised levels of serum nitric oxide metabolites are associated with a lower number of clinical relapses and a non-progressive disease course. These findings, although preliminary, pose interesting questions on the role of nitric oxide in the pathogenesis of MS. Inducible NO production in the early stages of the disease may be beneficial. PMID- 11499640 TI - The SOX10 transcription factor: evaluation as a candidate gene for central and peripheral hereditary myelin disorders. AB - The SOX10 transcription factor is involved in development of neural crest derivatives and fate determination in glial cells. SOX10 mutations have been found in patients with intestinal aganglionosis and depigmentation with deafness (Waardenburg-Hirschsprung). Associated neurological signs have been reported in some cases, including a patient exhibiting a central and peripheral myelin deficiency. Therefore, we screened for SOX10 mutations in a large cohort of patients with peripheral and central myelin disorders. 56 were affected by classical demyelinating Charcot-Marie-Tooth disease without identified mutations in the genes encoding PNS myelin proteins (PMP22, P0), connexin 32 and the zinc finger transcription factor, EGR2. 88 patients with undetermined leukodystrophy were selected from a large European prospective study. Associated clinical, magnetic resonance imaging and electrophysiological signs were consistent with a defect in CNS myelination in 83 and with an active degeneration of the CNS myelin in 5. No abnormalities in the proteolipid protein gene (PLP) were found. The absence of SOX100 mutation in this large cohort of patients suggests that this gene is not frequently involved in peripheral or central inherited myelin disorders. PMID- 11499641 TI - Polio survivors--well educated and hard working. AB - 243 patients were diagnosed with acute poliomyelitis (polio) in Western Norway between 1950 and 1954; 186 were paralytic and 57 non-paralytic. This study examines how polio influenced their education, employment, profession, annual income, marital status and energy for leisure activities. 149 of the patients identified were alive and 98 of the matched controls responded to a questionnaire. Education length did not differ between acute paralytic polio patients, acute non-paralytic polio patients and controls. Fifty percent of the patients with residual weakness and 77 % of the patients with normal muscle power were employed, against 73 % of the controls (P=0.014). A higher proportion of patients without motor deficits had manual work than those with weakness or controls (P=0.002). There was no significant association between severity of weakness and education, employment and profession. Physical ability had been an important factor for the choice of education and profession for all the polio patients, but not for controls (P < 0.001). Annual income did not differ significantly between patients and controls. Residual weakness increased the chance of being single (P=0.023), although as many as 79% had married. 53 % of the patients with weakness claimed that fatigue prevented hobbies, compared wich 31% of the other patients and only 16% of the controls (P < 0.001). There was no significant association between severity of weakness and fatigue. In conclusion, the polio patients are generally well educated, provide their own income and marry. However, their polio has influenced choice of education and profession, and polio patients with persisting weakness differ from controls and polio patients without motor deficits regarding employment and marital status. PMID- 11499642 TI - Ultrasonography and contrast-enhanced MRA in ICA-stenosis: is conventional angiography obsolete? AB - BACKGROUND: The standard techniques for the screening and staging of internal carotid artery (ICA) stenosis are Doppler (continuous wave) and Duplex sonography. For the imaging of ICA occlusive disease, magnetic resonance angiography (MRA) is replacing digital subtraction angiography (DSA). The purpose of this observational study was to assess whether contrast enhanced MRA (CE-MRA) combined with ultrasound provided sufficient information for the planning of surgical treatment. METHODS: CE-MRA was performed in 195 patients (mean age 67.5 years) with sonographic evidence of severe ICA stenosis. The MRA examination protocol contained a heavily T1-weighted contrast bolus enhanced 3D-gradient echo sequence. The degree of stenosis was estimated retrospectively by two experienced neuroradiologists who were blinded to the sonographic findings. RESULTS: The consistency of MRA and ultrasound was sufficient to plan thrombendarterectomy in 182/195 patients. The estimations of the degree of stenosis were congruent between MRA and ultrasound in 91% of 197 vessels with high-degree carotid artery stenosis. CE-MRA evaluation had a high interobserver agreement. In 3 cases ultrasound examination diagnosed a filiform ICA stenosis which was not visible with MRA. In all these cases, DSA and the intraoperative findings revealed very short (1-2 mm), high-grade, excentric stenosis. CE-MRA correctly detected patency in 5 patients with high-grade and low-flow carotid artery stenosis, which had been regarded as occluded by ultrasound. Conversely with, in CE-MRA two occluded vessels were falsely considered as open. CONCLUSION: The combination of sonography and CE-MRA is a powerful tool for the non-invasive presurgical evaluation of the carotid arteries. DSA should be reserved for selected cases. PMID- 11499643 TI - Cervico-thoracic disc protrusions in controlled compressed-air diving: clinical and MRI findings. AB - Prevalence of cervical and thoracic disc protrusions was investigated by MRI in 24 military long-term compressed-air divers and 24 controls. A total of 26 disc protrusions (17 cervical disc protrusions) were detected in 58% of the divers whereas 18 protrusions were detected in 38 % of the controls (13 disc cervical protrusions). There was no significant difference between groups and no correlation with the diving experience. Neurological examination revealed no clinical abnormalities. In contrast to a recent study, our results suggest that long-term divers are at no increased risk for accumulating spinal disc protrusions or intramedullary abnormalities. PMID- 11499644 TI - T1 hypointensity of the spinal cord in multiple sclerosis. AB - It has recently been shown in multiple sclerosis (MS) that the volume of T1 hypointense lesions in the brain explains more of the variance in disability amongst patients than T2 lesion volume. T1 hypointense lesions may therefore represent areas of underlying pathology likely to be of functional significance, such as axonal loss. The spinal cord is a common area of involvement in MS and its dysfunction is likely to be responsible for much of the motor disability seen. Hence it serves as a useful model by which to examine the functional relevance of differing imaging sequences. We have therefore examined the relationship between T1 signal intensity in the spinal cord and disability in 60 patients with MS. We have also examined the relationship between T1 signal intensity and atrophy of the cord, as the latter is another potential marker of axonal loss. Sixty patients with MS underwent spinal cord imaging with a T1 weighted sequence to acquire axial sections of the cord at the C2 level. These sections were histogram matched to allow comparison of image intensity and a manual outlining technique was applied from which the mean cord intensity was calculated. Within the patient group there was a significant relationship between T1 signal intensity and disability as measured with the EDSS (r = -0.4, p < 0.005) and also between T1 signal intensity and atrophy (r = 0.36, p < 0.005). This study demonstrates that disability and atrophy are associated with a generalised reduction in cord signal on T1 weighted images. A lower T1 signal intensity in the spinal cord may be more pathologically specific than T2 hyperintensity and may represent underlying axonal loss, although gliosis and predominant white matter atrophy are alternative possibilities. PMID- 11499645 TI - Sarcoidosis presenting as rapidly progressive dementia: clinical and neuropathological evaluation. PMID- 11499646 TI - Systemic vasculitis and hyperthyroidism in a patient with myasthenia gravis. PMID- 11499647 TI - Tracheostomy scars in Guillain-Barre syndrome: a reason for concern? PMID- 11499648 TI - Fat embolism syndrome: new MRI findings. PMID- 11499649 TI - Paroxysmal hypertensive crises induced by selegiline in a patient with Parkinson's disease. PMID- 11499650 TI - Extracorporeal photochemotherapy reduces the incidence of relapses in experimental allergic encephalomyelitis in DA rats. PMID- 11499651 TI - Cortical-subcortical lesions in "reversible posterior leukoencephalopathy syndrome". Encephalopathy or seizures? PMID- 11499652 TI - Camillo Golgi (1843-1926). PMID- 11499653 TI - Short- and long-term efficacy of oral cyclophosphamide and steroids in patients with membranous nephropathy and renal insufficiency. Study Group. AB - BACKGROUND: Up to half of the patients with idiopathic membranous nephropathy (iMN) will develop renal failure. Preferably, immunosuppressive treatment should be restricted to patients at risk for the development of end-stage renal disease. However, the evidence that immunosuppressive treatment is effective in patients with iMN and renal insufficiency is weak and based on few studies with short follow-up in a limited number of patients. METHODS: We have analyzed the efficacy of immunosuppressive treatment in a large number of patients with membranous nephropathy and renal insufficiency. Since 1991, we have prospectively treated 39 patients (31 M, 8 F) with membranous nephropathy and evidence of deterioration of renal function. Treatment consisted of oral cyclophosphamide, 1.5-2.0 mg/kg body weight for 12 months, and corticosteroids for 6 months. At regular intervals blood pressure, serum creatinine, serum albumin, and proteinuria were measured. Adverse events were recorded. RESULTS: Average follow-up is 32 months (range 6 104), 18 patients have been followed for more than 3 years. Mean age of the patients was 55 +/- 12 years. In the 6 months before start of therapy, serum creatinine increased from 150 +/- 74 to 226 +/- 108 micromol/l. After start of treatment renal function rapidly improved, serum creatinine at 12 months averaging 143 +/- 62 micromol/l. Proteinuria decreased from 10.3 +/- 4.9 g/10 mmol creatinine at baseline to 2.2 +/- 2.4 g/10 mmol creatinine at month 12. These initial favorable effects have persisted. Overall, 12 patients have developed a complete remission of proteinuria (persistent in 11), and an additional 19 have developed a partial remission of proteinuria (persistent in 15). Thus far, only one treated patient has developed end-stage renal disease. Side effects are a major drawback of the treatment, with 7 patients being admitted, mainly for the treatment of infectious complications. CONCLUSIONS: Cyclophosphamide is effective in the treatment of patients with idiopathic membranous nephropathy and deterioration of renal function. The favorable effects are maintained well beyond the one-year treatment period. Therefore, we propose that in patients with iMN immunosuppressive therapy can be restricted to patients at high risk for end-stage renal disease. PMID- 11499654 TI - Factors predictive of renal involvement in patients with primary Sjogren's syndrome. AB - AIM: To identify clinical and immunological risk factors underlying the development of renal involvement in primary Sjogren's syndrome (pSS). SUBJECTS AND METHODS: Seventy-eight patients (75 females, 3 males) with pSS were carefully interviewed and clinical and laboratory data from the time of diagnosis recorded. The baseline data on patients shown to have either latent or overt distal renal tubular acidosis (dRTA), mild proteinuria or increased urinary excretion of alpha 1 microglobulin (alpha1m) after a mean disease duration of 9 +/- 4 years, were compared to the baseline data on those who did not have these manifestations at follow-up. RESULTS: Patients with subsequent latent or overt dRTA were found to have significantly higher baseline levels of serum total gamma-globulin (24 +/- 7 vs. 19 +/- 6 g/l, p = 0.011) and serum protein (84 +/- 7 vs. 79 +/- 7 g/l, p = 0.024) compared to those with normal renal acidification capacity. The baseline levels of serum beta-2 microglobulin (beta2m) were higher in patients with an acidification defect than in those with normal acidification capacity (3.1 +/- 1.1 vs. 2.6 +/- 0.8 mg/l, p = 0.072). In those with subsequent proteinuria the levels of serum beta2m were almost significantly higher at baseline as compared to those with normal urinary protein excretion (3.1 +/- 1.4 vs. 2.5 +/- 0.8 mg/l, p = 0.052). The subgroup of pSS patients who had increased urinary alpha1m excretion as a sign of tubular proteinuria, had higher baseline levels of ESR (55 +/- 27 mm/h vs. 40 +/- 23 mm/h, p = 0.076) and significantly higher baseline levels of serum beta2m (4.6 +/- 1 .8 vs. 2.6 +/- 0.8 mg/l, p = 0.029) as compared to those with normal urinary alpha1m excretion. CONCLUSIONS: High levels of serum total gamma-globulin, serum protein and serum beta2m were the best predictors of the development of dRTA in pSS patients. High baseline levels of serum beta2m were also associated with the subsequent occurrence of mild proteinuria and increased urinary alpha1m excretion in patients with pSS. PMID- 11499655 TI - Renal biopsy in elderly patients: clinicopathological correlation in 117 Korean patients. AB - AIM: In the elderly with renal disease, the clinical presentations are frequently inconsistent with the pathologic findings. We tried to clarify the differences in pathological findings between the young and the elderly, in Korea and in Western countries, and the usefulness of a percutaneous renal biopsy in the elderly with renal disease. PATIENTS AND METHODS: We analyzed the clinical presentations and spectrums of renal histopathology by reviewing medical records and renal biopsy reports retrospectively in 117 Korean patients aged 60 years or more with renal disease. RESULTS: 85 patients had primary renal disease. The remaining 32 patients had renal diseases associated with systemic conditions. Out of the 85 patients with primary renal disease, 61 cases presented as idiopathic nephrotic syndrome. Compared with renal biopsy results of younger adult patients (age 15 59, n = 1,908), membranous nephropathy, crescentic glomerulonephritis, membranoproliferative glomerulonephritis, amyloidosis, light chain disease, and thrombotic thrombocytopenic purpura were more prevalent, but IgA nephropathy and lupus nephritis were less common in the elderly patients. In clinical presentation, nephrotic syndrome and rapidly progressive renal failure were more prevalent, but asymptomatic urinary abnormality was less common in elderly patients. The responsiveness to treatment was good in elderly patients with minimal-change lesion (complete remission in all patients) but poor in crescentic glomerulonephritis, IgA nephropathy, and membranoproliferative glomerulonephritis. From the above findings, the clinical presentation, patterns of histopathology and responsiveness to treatment of elderly Korean patients were similar to those of the younger Korean control group and the Western elderly group. CONCLUSION: Percutaneous renal biopsy is a useful diagnostic aid and can be used as a therapeutic guideline even in elderly patients with renal disease. PMID- 11499656 TI - Early changes of bone histology and circulating markers of bone turnover after parathyroidectomy in hemodialysis patients with severe hyperparathyroidism. AB - AIMS: There have so far been no reports on the changes in bone histology in the early period after parathyroidectomy and autografting (PTX-AG). We investigated the effects of PTX-AG on bone histology during the initial 12 weeks after undergoing these surgical procedures. MATERIALS AND METHODS: We performed bone histomorphometry 3 times (before as well as 4 and 12 weeks after PTX-AG) in 6 patients and 2 times (before and 4 weeks after PTX-AG) in 3 hemodialysis patients. In addition, the circulating parameters of bone metabolism were also assessed before and after PTX-AG in all 9 patients. The changes in the histomorphometric (static) parameters between pre-surgery and 4 weeks after surgery and those between 4 weeks and 12 weeks after surgery were assessed by the t-test while changes in the circulating parameters of bone metabolism were analyzed by Friedman's test. RESULTS: Bone formation parameters including carboxy terminal propeptide of human type I procollagen (PICP), alkaline phosphatase (ALP) and intact osteocalcin (i-OC) were all extremely high before surgery. These parameters initially increased after PTX-AG and thereafter gradually declined. In contrast, the circulating bone resorption parameters including tartrate-resistant acid phosphatase (TRAP) and deoxypyridinoline (Dpyr) were also extremely high at baseline but markedly declined after operation. Osteoid-related parameters including osteoid volume (OV/BV), osteoid surface (OS/BS), and osteoid thickness (O.Th) all initially increased at 4 weeks after PTX-AG. In contrast, osteoblast surface (Ob.S/BS), osteoclast surface (Oc.S/BS), eroded surface (ES/BS), and fibrosis volume (Fb.V/TV) all decreased at 4 weeks after surgery, while Ob.S/BS decreased further at 12 weeks in cases 1-6. Although bone mineralization was ongoing at 4 weeks after surgery, both the mineral apposition rate (MAR) and bone formation rate (BFR) remained below the mean for normal individuals. CONCLUSIONS: The circulating bone formation parameters and osteoid-related parameters showed an initial increase after PTX-AG. The concomitant decline in the circulating bone resorption parameters reflected the reduction in bone resorption. BFR decreased, but bone mineralization did not stop after PTX-AG. PMID- 11499657 TI - Parathyroid hormone changes during phosphorus load in patients with chronic renal insufficiency with low serum parathyroid hormone or adynamic bone disease. AB - Adynamic bone disease (ABD) is frequently associated with low serum parathyroid hormone (PTH) concentrations. Many clinical and therapeutic conditions have been associated with ABD, and recently, a low phosphorus intake accompanied by low serum concentration of phosphorus and PTH has been described. AIM: To evaluate the parathyroid gland response of chronic renal insufficiency patients (CRI) with low serum PTH or ABD to a phosphorus load. METHODS: We examined the effects of 0.5 and 1.0 g/d of phosphorus load over a period of 60 days in 18 patients with mild CRI with a bone biopsy showing ABD (n = 7) or with low serum PTH (serum intact PTH < or = 40 ng/l) and serum phosphorus < 4.5 mg/dl (n = 11). RESULTS: Serum intact PTH increased significantly only after 1 g of phosphorus (58.5 to 83 ng/l) with a median percent increase of 72%. PTH secretion increased more in patients with lower basal PTH levels (81%). Serum phosphorus did not change significantly and urinary phosphorus increased from 487 to 1,062 mg/dl (p < 0.05). Significant decreases in serum ionized calcium (from 1.26 to 1.19 mmol/l) and calcitriol (from 34.5 to 24.9 pg/ml) were observed. Changes in PTH were inversely correlated with changes in serum ionized calcium (r = -0.54, p < 0.05) and the final PTH concentrations were positively correlated with changes in serum phosphorus (r= 0.52, p < 0.05). CONCLUSIONS: The parathyroid glands of chronic renal insufficiency patients with "relative hypoparathyroidism" or ABD responded to a phosphorus load with an increase in serum PTH levels. The decrease in serum ionized calcium and calcitriol as well as minimal changes in serum phosphorus appeared to be involved in this response. PMID- 11499658 TI - Cardiac troponins have no prognostic value for acute and chronic cardiac events in asymptomatic patients with end-stage renal failure. AB - BACKGROUND: Cardiovascular diseases determine overall mortality in patients with end-stage renal failure. Therefore, testing for myocardial ischemia is important. Elevation of cardio-specific troponins have been frequently measured in patients with end-stage renal failure. Thus, we studied systematically whether patients on chronic intermittent hemodialysis without overt coronary heart disease have increased serum levels of cardiac troponin T and cardiac troponin I. After 2 years, the patients were screened again for cardiac events. METHODS AND RESULTS: The patients had no history of angina during the previous 3 months or myocardial infarction (MI) within the previous 2 years. For analysis we used two cardio specific assays for troponin T as well as for troponin I and compared the results with the CK-MB concentration. In a number of patients serum concentrations were elevated above the reference range as follows: troponin T rapid bedside assay: 41 of 100 patients, troponin I rapid bedside assay: 27 of 100 patients, quantitative measurement oftroponin T: 22 of 100 patients, quantitative measurement oftroponin I: 7 of 100 patients, CK-MB: 2 of 100 patients. The increased serum levels of cardiac troponins were neither the result of uremic perimyocarditis (pericardial effusion), changes in the hemodialysis regimen, pulmonary congestion nor were they consistent with the etiology of renal failure. None of the patients with an elevated troponin level in either of the test suffered from any acute cardiac event initially. Within 2 years 18 of 100 patients died, 13 out of them because of cardiac events. Fourteen patients had a myocardial infarction and 19 patients developed angina pectoris. Sensitivity and specificity (0.75 and 0.67) of troponin T rapid bedside assay for MACE (angina pectoris, MI, cardiac death) was lower compared to studies in patients with normal renal function. Correlation between troponin elevation and late outcome was low or absent. CONCLUSION: Patients on chronic intermittent hemodialysis frequently present with elevated TnT and TnI levels which cannot be used as predictors of acute and chronic cardiac events. Rapid bedside assays have a lower specificity than quantitative assays. PMID- 11499659 TI - Dialysis membrane-induced neutrophil apoptosis is mediated through free radicals. AB - Patients on hemodialysis are prone to infection. Neutrophils are the host's first line of defense against certain invading pathogenic microorganisms. Since apoptotic neutrophils are functionally compromised we examined the effect of dialysis membranes on neutrophil apoptosis. Dialysis patients showed greater (p < 0.001) neutrophil apoptosis when compared with control subjects. Cellulose acetate membranes directly promoted (p < 0.001) neutrophil apoptosis. Cellulose acetate membrane-treated neutrophils exhibited greater apoptosis (p < 0.01) when compared with polysulfone membrane-treated neutrophils. Superoxide dismutase (SOD) partly inhibited the cellulose acetate membrane-induced neutrophil apoptosis, whereas both catalase and dimethylthiourea (DMTU) inhibited the polysulfone membrane-induced neutrophil apoptosis. Similarly, L-NAME, a nitric oxide synthase inhibitor, attenuated both the cellulose acetate and the polysulfone membrane-induced neutrophil apoptosis. In addition, cellulose acetate and monocyte interaction products promoted (p < 0.001) neutrophil apoptosis. These results suggest that dialysis membranes can promote neutrophil apoptosis directly as well as through their interaction with monocytes. The direct effect of dialysis membranes seems to be mediated partly through the generation of reactive oxygen species. PMID- 11499660 TI - Amphotericin B, mercury chloride and peritoneal transport in rabbits. AB - BACKGROUND: The effect of glucose-induced ultrafiltration in peritoneal dialysis is dependent on the presence and function of ultrasmall transendothelial cell water channels. The mercury-sensitive aquaporin-1 was thought to represent these transcellular pores. Amphotericin B (ampho B) has been reported to increase ultrafiltration in both experimental and patient studies. The objective of this study was to investigate the hypothesis that intraperitoneal ampho B increases and mercury chloride inhibits aquaporin-1-mediated water transport in a chronic peritoneal dialysis model in the rabbit. MATERIAL AND METHODS: Eighteen female New Zealand White rabbits were included for peritoneal catheter implantation. Peritoneal transport parameters were determined in all rabbits by standard peritoneal permeability analysis (SPAR) with 3.86% glucose-based dialysis solution during a one-hour dwell prior the intervention SPARs, as a control. Ampho B (0.06 mg/kg body weight) was added to the dialysate for 3 (n = 9) or 5 consecutive days (n = 5) before investigation. Four rabbits were investigated after 3-day i.p. 0.6 mg/kg body weight ampho B. In 3 rabbits 0.06 mg/kg body weight liposomal ampho B was administered i.p. during 3 days before intervention SPAR. Fifteen rabbits were investigated during a one-hour dwell with 0.1 mM HgCl2 containing 3.86% glucose-based dialysis solution, while they were anesthetized. Three of these underwent in vivo fixation with glutaraldehyde prior to the HgCl2 SPAR to prevent toxic effects of mercury on peritoneal tissues. RESULTS: Intraperitoneal administration of ampho B did enhance the change in intraperitoneal volume during a one-hour dwell after 3-day i.p. treatment with the low dose (p < 0.02), but it did not affect peritoneal solute permeability. This was likely mediated by transcellular water channels, but not by aquaporin-1. No beneficial effects on the ultrafiltration were found with prolonged treatment or with the higher dose. Ultrafiltration decreased (8 ml/4 h to 1 ml/4 h, p < 0.03) after i.p. administration of HgCl2 with and without in vivo fixation, accompanied by a significant decrease in aquaporin-mediated water transport, estimated as the sieving of sodium (p < 0.001). Marked increases in the clearances of macromolecules were found after i.p. HgCl2 administration due to toxic effects: total protein clearance from 97 to 172 microl/min, p < 0.005, and albumin clearance from 59 to 158 microl/min, p < 0.005. These changes were less pronounced after in vivo fixation. CONCLUSION: Ampho B has likely no clinical relevance in treatment of ultrafiltration failure in PD patients. Aquaporin mediated water transport could be inhibited and consequently ultrafiltration was reduced by i.p. administration of mercury chloride in our rabbit model. PMID- 11499661 TI - Non-diabetic nodular glomerulosclerosis associated with p-ANCA seropositivity: a case report and review of the literature. AB - A 73-year-old white man with slowly progressive chronic renal failure and nephrotic-range proteinuria was found to have antineutrophil cytoplasmic antibody in a perinuclear pattern (p-ANCA) at a titer of 1:800. Renal histologic findings revealed an advanced scarring glomerulopathy with diffuse and nodular mesangial sclerosis. Light, electron, and immunofluorescence microscopic findings were highly suggestive of diabetic glomerulosclerosis. Interestingly, this patient had no history of diabetes mellitus or diabetic retinopathy. The presence of p-ANCA positivity can be found in patients with a broad range of renal histologic findings, and does not necessarily imply the existence of pauci-immune necrotizing crescentic glomerulonephritis. For this reason, we urge caution in the empiric cytotoxic treatment of p-ANCA-associated renal disease in stable patients. When possible, a tissue diagnosis should be made. PMID- 11499662 TI - Focal segmental glomerulosclerosis associated with acromegaly. AB - Experimental studies in rats have demonstrated an association between focal segmental glomerulosclerosis (FSGS) and growth hormone, but patients with FSGS complicating acromegaly are very rare. In this report we present a case of FSGS associated with acromegaly. With a long history of soft tissue swelling of hands and feet, elevated plasma growth hormone levels and other biochemical abnormalities, a 53-year old male had suffered from acromegaly for over 15 years. He had moderate proteinuria for 6 years, but never evidenced nephrotic syndrome. A renal biopsy specimen revealed FSGS and glomerular hypertrophy. Trans sphenoidal surgical removal of the pituitary adenoma resulted in the normalization of elevated growth hormone and insulin-like growth factor I levels, but proteinuria continued. This case suggests that the overproduction of growth hormone may participate, at least in part, in the development of human FSGS. It is possible that once FSGS is present in an acromegalic patient, cessation of GH overproduction may not be enough to reverse it. PMID- 11499663 TI - Horner's syndrome secondary to internal jugular catheterization. AB - We present three cases of people who developed Horner's syndrome following repeated attempts for catheterization of the internal jugular vein for hemodialysis. Any other neurologic finding or evidence of mass lesions in the neck or pulmonary apex could not be determined. Horner's syndrome should be considered a possible complication of percutaneous internal jugular vein catheterization for hemodialysis. PMID- 11499664 TI - Effect of acute urinary retention on glomerular filtration rate. PMID- 11499665 TI - Small vessel vasculitis with crescentic glomerulo-nephritis associated with IgA lambda monoclonal gammapathy. PMID- 11499666 TI - Creatininemia during fibrate therapy: a reply to Lipscombe et al. PMID- 11499667 TI - RPA position on public disclosure of information. Renal Physicians Association. PMID- 11499669 TI - Evaluation of the fragile X (FRAXA) syndrome with methylation-sensitive PCR. AB - The fragile X (FRAXA) syndrome is the most common form of inherited mental retardation in males. Its peculiar pattern of inheritance results from the parent of origin-specific expansion of a CGG-repeat within the FMR1 gene on the X chromosome. In patients, gene function is abolished by hypermethylation of the promoter and the massively expanded repeat. We have developed a methylation sensitive polymerase chain reaction (MS-PCR) strategy that combines repeat-length and methylation analysis of the CGG-repeat and the promoters of the FMR1 and XIST genes. The allelic methylation of the latter opposes that of the FMR promoter and serves as an internal control and standard for semiquantitative analyses. This system enables the delineation of 11 distinct patterns encountered in nonaffected, carrier, and affected males and females. We have evaluated our system on well-defined samples with different FMR1 mutations and have used it for the diagnostic evaluation of 253 male and 80 female probands. In the male group, we have identified five full mutations, and three gray-zone and premutation alleles with 54, 55, and 62 repeats, respectively. The female group consists of 33 normal homozygote and 41 heterozygote individuals, two of whom harbor a gray zone allele with 47 repeats, none with a premutation, and six with a full mutation. Our MS-PCR approach allows the currently most comprehensive diagnostic evaluation of the FRAXA syndrome in a cost- and time-efficient fashion. In addition, it is a valuable tool for the analysis of clonality and skewing phenomena in females. PMID- 11499668 TI - Distribution of glucose-6-phosphate dehydrogenase mutations in Southeast Asia. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a heterogeneous enzyme abnormality with high frequency in tropical areas. We performed population screening and molecular studies of G6PD variants to clarify their distribution and features in Southeast Asia. A total of 4317 participants (2019 males, 2298 females) from 16 ethnic groups in Myanmar, Lao in Laos, and Amboinese in Indonesia were screened with a single-step screening method. The prevalence of G6PD-deficient males ranged from 0% (the Akha) to 10.8% (the Shan). These G6PD deficient individuals and 12 G6PD-deficient patients who had been diagnosed at hospitals in Indonesia and Malaysia were subjected to molecular analysis by a combination of polymerase-chain-reaction-based single-strand conformation polymorphism analysis and direct sequencing. Ten different missense mutations were identified in 63 G6PD-deficient individuals (50 hemizygotes, 11 heterozygotes, and 2 homozygotes) from 14 ethnic groups. One missense mutation (1291 G-->A) found in an Indonesian Chinese, viz., G6PD Surabaya, was previously unknown. The 487 G-->A (G6PD Mahidol) mutation was widely seen in Myanmar, 383 T- >C (G6PD Vanua Lava) was specifically found among Amboinese, 871 G-->A (G6PD Viangchan) was observed mainly in Lao, and 592 C-->T (G6PD Coimbra) was found in Malaysian aborigines (Orang Asli). The other five mutations, 95 A-->G (G6PD Gaohe), 1003 G-->A (G6PD Chatham), 1360 C-->T (G6PD Union), 1376 G-->T (G6PD Canton), and 1388 G-->A (G6PD Kaiping) were identified mostly in accordance with distributions reported previously. PMID- 11499670 TI - Analyses of genetic abnormalities in type I CD36 deficiency in Japan: identification and cell biological characterization of two novel mutations that cause CD36 deficiency in man. AB - To elucidate genetic abnormalities in type I CD36 deficiency, we analyzed 28 Japanese subjects whose platelets and monocytes/macrophages lacked CD36 on their surface. We identified two novel mutations in the CD36 gene. One was a complex deletion/insertion mutation, in which 3 bp, GAG, were deleted at nucleotide (nt) 839-841, and 5 bp, AAAAC, were inserted at the same position (839-841del- >insAAAAC). Mutation 839-841del-->insAAAAC led to a frameshift and appearance of a premature stop codon; it was also accompanied with a marked reduction in the amount of CD36 mRNA. The other was a 12-bp deletion at nt 1438-1449 (1438 1449del) accompanied with or without skipping of exon 9 (nt 959-1028). Mutation 1438-1449del led to an inframe 4-amino-acid deletion, whereas exon 9 skipping led to a frameshift and the appearance of a premature stop codon. Expression assay revealed that both 1438-1449del and exon 9 skipping directly caused impairment of the surface expression of CD36. A survey of the five known mutations including 839-841del-->insAAAAC and 1438-1449del in type I CD36-deficient subjects demonstrated that the five mutations covered more than 90% of genetic defects among them and that the substitution of T for C at nt 478 (478C-->T) was the most common mutation with more than 50% frequency. However, none of the four subjects that possessed isoantibodies against CD36 had 478C-->T, suggesting that 478C-->T prevents the production of isoantibodies against CD36. PMID- 11499671 TI - The structure of duplications on human acrocentric chromosome short arms derived by the analysis of 15p. AB - We report the molecular analysis of a 130-kb DNA region containing a junction between beta and non-beta satellite DNA from chromosome 15p. The genomic region is characterized by beta satellite blocks intermingled with variants of the D4Z4 repeat, and duplicons from 4q24 and 4q35. Besides the p-arm of acrocentric chromosomes, the duplicons showed a wide genomespread involving pericentromeric, sub-telomeric, and interstitial regions. In this regard, the paralogous sequences were characterized by a high similarity index (96%), thus indicating a recent transposition during the evolution. The acrocentrics differedwith regard to the location of the 4q24 paralogous region, since it mapped on the p-arm of chromosomes 13-15 and 21, but only on 22q11.2. Conversely, the 4q35 duplication marked the p-arm of all the acrocentrics. In different individuals, the short arm of acrocentric chromosomes revealed a great variability of sequence representation and location at p11 and/or p13 for both the 4q24 and 4q35 duplications. The studied genomic region from chromosome 15p, of which a contig of approximately 200 kb has been derived, could lead to more detailed investigations into the sequence organization and possible biological function of chromosome regions that are located close to the rDNA array. PMID- 11499672 TI - Improved definition of chromosomal breakpoints using high-resolution multicolour banding. AB - Characterisation of chromosome rearrangements using conventional banding techniques often fails to determine the localisation of breakpoints precisely. In order to improve the definition of chromosomal breakpoints, the high-resolution multicolour banding (MCB) technique was applied to identify human chromosome 5 breakpoints from 40 clinical cases previously assessed by conventional banding techniques. In 30 cases (75%), at least one breakpoint was redefined, indicating that MCB markedly improves chromosomal breakpoint localisation. The MCB pattern is highly reproducible and, in contrast to conventional banding pattern, is consistent in both short and elongated chromosomes. This might be of fundamental interest for the detection of chromosomal abnormalities, especially in tumour cells. Moreover, MCB even allows the detection of abnormalities that remain cryptic in GTG-banding analysis. PMID- 11499673 TI - Statistical estimation and pedigree analysis of CCR2-CCR5 haplotypes. AB - As more SNP marker data becomes available, researchers have used haplotypes of markers, rather than individual polymorphisms, for association analysis of candidate genes. In order to perform haplotype analysis in a population-based case-control study, haplotypes must be determined by estimation in the absence of family information or laboratory methods for establishing phase. Here, we test the accuracy of the Expectation-Maximization (EM) algorithm for estimating haplotype state and frequency in the CCR2-CCR5 gene region by comparison with haplotype state and frequency determined by pedigree analysis. To do this, we have characterized haplotypes comprising alleles at seven biallelic loci in the CCR2-CCR5 chemokine receptor gene region, a span of 20 kb on chromosome 3p21. Three-generation CEPH families (n=40), totaling 489 individuals, were genotyped by the 5'nuclease assay (TaqMan). Haplotype states and frequencies were compared in 103 grandparents who were assumed to have mated at random. Both pedigree analysis and the EM algorithm yielded the same small number of haplotypes for which linkage disequilibrium was nearly maximal. The haplotype frequencies generated by the two methods were nearly identical. These results suggest that the EM algorithm estimation of haplotype states, frequency, and linkage disequilibrium analysis will be an effective strategy in the CCR2-CCR5 gene region. For genetic epidemiology studies, CCR2-CCR5 allele and haplotype frequencies were determined in African-American (n=30), Hispanic (n=24) and European-American (n=34) populations. PMID- 11499674 TI - Preimplantation genetic diagnosis for spinal and bulbar muscular atrophy (SBMA). AB - X-linked spinal and bulbar muscular atrophy is characterized by adult onset motor neuron disease and results from a defect in the androgen receptor. The disease is caused by a dynamic mutation in the first exon of the androgen receptor gene, involving a CAG trinucleotide repeat. We have developed a single-cell polymerase chain reaction assay for the androgen receptor gene and describe the application of this assay for preimlantation genetic diagnosis (PGD) in a couple at risk, where the female partner is a carrier of 47 repeats. Diagnosis was based on the detection of both normal and expanded alleles. Allele dropout of the expanded allele was observed in only 1 of 25 lymphoblasts of the carrier and of a non expanded allele in 1 of 20 research blastomeres tested before the actual PGD. One contraction of four repeats was also found in the carrier's lymphoblasts. Neither expansions nor contractions were observed in the blastomeres biopsied from 11 embryos. Two embryos were unaffected, eight were female carriers and one was an affected male embryo. PMID- 11499675 TI - Acrocentric chromosome disomy is increased in spermatozoa from fathers of Turner syndrome patients. AB - The aim of the present study was to investigate whether there was an increase of aneuploidy in the sperm from fathers of Turner syndrome patients of paternal origin who, in a previous study, showed an elevated incidence of XY meiotic nondisjunction. Sperm disomy frequencies for chromosomes 4, 13, 18, 21 and 22 were assessed by fluorescence in situ hybridisation in four of these individuals. As a group, the Turner syndrome fathers showed a general increase in disomy frequencies for chromosomes 13, 21 and 22, with a statistically significant increase in disomy frequencies for chromosomes 13 and 22 in one of the fathers and for chromosome 21 in two of them. Data from a previous work carried out by us in two fathers of Down syndrome patients of paternal origin also revealed increased sperm disomy frequencies for chromosomes 13, 21 and 22. Pooled as one group, these six fathers of aneuploid offspring of paternal origin had a statistically significant increase in the frequency of nondisjunction for these chromosomes with respect to control individuals. Our findings indicate that there may be an association between fathering aneuploid offspring and increased frequencies of aneuploid spermatozoa. Such increases do not seem to be restricted to the chromosome pair responsible for the aneuploid offspring. Acrocentric chromosomes and other chromosome pairs that usually show only one chiasma during meiosis seem to be more susceptible to malsegregation. PMID- 11499676 TI - A 1.1-kb duplication in the p67-phox gene causes chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is a rare inherited immunodeficiency that is caused by a functional defect of the NADPH oxidase of phagocytes, and that leads to severe recurrent infections. CGD results from the absence or the dysfunction of various components of NADPH oxidase, and autosomal recessive CGD with the lack of p67-phox (A67 CGD) is the rarest form of the disease. Identifying familiar mutations in subjects with A67 CGD provides the most reliable method of detecting carriers and is the basis for prenatal diagnosis. In the present study, we report the detailed characterization of the first duplication in the p67-phox gene identified in a 30-year-old patient affected by systemic aspergillosis attributable to p67-phox deficiency. We show that this new mutation involving exons 9 and 10 is the result of a tandem duplication of approximately 1.1 kb, which resulted from the juxtaposition of intron 8 to intron 10. We have sequenced both the junction fragment of this duplication and the corresponding wild-type regions and have found that the breakpoint regions in intron 8 and in intron 10 show limited homology. Our result suggests that this interchange arose as an illegitimate recombination event. As in other non-homologous rearrangements previously reported, the duplication breakpoints are located within the sequence motif 5'-CCAG-3' and its complement 5'-CTGG-3'. PMID- 11499677 TI - Partial maternal heterodisomy of chromosome 17q25 in a case of severe mental retardation. AB - We report a segmental maternal uniparental heterodisomy of chromosome 17 (mat UPD17) in a 3-year-old boy presenting with hyperactivity, major instability, mental retardation and facial dysmorphism. Since conventional and high resolution karyotypes were normal, this patient was tested for cryptic telomeric rearrangements by using the recently developed fluorescent genotyping-based technology. The mat UPD17 segment extended for a small 11-cM region of the distal chromosome 17q. Trisomy 17 in circulating lymphocytes and skin fibroblasts was excluded. Our finding emphasizes the potential use of fluorescent genotyping to detect uniparental disomies and suggests that chromosome 17q25 should contain one or several imprinted genes of particular importance for brain development. PMID- 11499678 TI - Interaction between the melanocortin-1 receptor and P genes contributes to inter individual variation in skin pigmentation phenotypes in a Tibetan population. AB - The melanocortin-1 receptor (MC1R) and P gene product are two important components of the human pigmentary system that have been shown to be associated with red hair/fair skin and cause type II oculocutaneous albinism, respectively. However, their contribution to inter-individual variation at the population level is not well defined. To this end, we genotyped 3 single nucleotide polymorphisms (SNPs) in the MC1R gene (Arg67Gln, Gln163Arg, Val92Met) and 2 SNPs in the P gene (IVS 13-15 and Gly780Gly) in 184 randomly ascertained Tibetan subjects, whose skin color was measured as a quantitative trait by reflective spectroscopy. Single locus analyses failed to demonstrate an association between any of the 5 SNPs and skin pigmentation. However, when an epistatic model was applied to the data, a significant gene-gene interaction was identified between Val92Met in MCIR and IVS13-15 in the P gene (F=2.43; P=0.0105). We also discuss the possible mechanisms of how gene interactions arise in signal transduction pathways. PMID- 11499679 TI - Genome-wide distribution of linkage disequilibrium in the population of Palau and its implications for gene flow in Remote Oceania. AB - Linkage disequilibrium (LD) between alleles on the same human chromosome results from various evolutionary processes and is thus telling about the history of populations. Recently, LD has garnered substantial interest for its value to map and fine-map disease genes. We examine the distribution of LD between short tandem repeat alleles on autosomes and sex chromosomes in the Remote Oceanic population of Palau to evaluate whether the data are consistent with a recent hypothesis about the origins of genetic variation in Palau, specifically that the population experienced extensive male-biased gene flow following initial settlement. Consistent with evolutionary theory based on effective population size, LD between X-linked alleles is stochastically greater than LD between autosomal alleles, however, small but detectable LD occurs for autosomal markers separated by substantial distances. By contrast, while Y-linked alleles experience only one-third the effective population size of X-linked alleles, their mean value for pairwise LD is only slightly larger than X-linked alleles. For a small population known to experience at least two extreme bottlenecks, 56 six-locus Y haplotypes exhibit remarkable diversity (0.96), comparable to Y diversity of Europeans, however, autosomal and X-linked markers display significantly less diversity, as measured by heterozygosity (4.1% less). Palauan Y haplotypes also fall into distinct clusters, again unlike that of Europe. We argue these data are consistent with waves of male-biased gene flow. PMID- 11499680 TI - The genes for the human VPS10 domain-containing receptors are large and contain many small exons. AB - The two human proteins with a VPS10 domain, SorLA and sortilin, both bind neuropeptides. Searching for other VPS10-domain proteins in the database revealed three new putative human neuropeptide receptors. The new receptors were designated SorCS1, SorCS2 and SorCS3, due to their identical domain composition, which, except for the N-terminal VPS10 domain, differs from that of SorLA and sortilin. Using the databases of the human genome project we elucidated the exon intron structures of the human VPS10-receptor genes. They contain many short exons, separated by introns, several of which extend over more than 50 kb. The three SorCS genes encompass more than 500 kb of genomic DNA and therefore represent some of the largest known human genes. All these genes map to chromosomal localisations of known genetic diseases, many of them neurological disorders, corresponding to the strong expression of these receptors in the brain. CpG islands are located in the first exon of each of the VPS10-receptor genes and might be involved in developmental or tissue-specific regulation of gene expression. PMID- 11499681 TI - Physical and transcriptional mapping of the X-linked cleft palate and ankyloglossia (CPX) critical region. AB - Cleft palate most commonly occurs as a sporadic multifactorial disorder with a clear but difficult to define genetic component. As a semi-dominant disorder, X linked cleft palate (CPX) provides a useful model to investigate a congenital defect that is little influenced by non-genetic factors. By using an Icelandic kindred, CPX has been localised between DXS1196 and DXS1217 and mapped, in a 3-Mb yeast artificial chromosome contig, at Xq21.3. Markers generated from this physical map have now been used to construct a contig of P1 and bacterial artificial chromosome clones for genomic DNA sequencing. Genomic DNA sequence analysis has revealed two novel expressed genes and two pseudogenes in the order Cen-KLHL4-LAMRL5-CAPZA1P-CPXCR1-Tel. KLHL4 and CPXCR1 are widely expressed in fetal tissues, including the tongue, mandible and palate. DNA mutation screening of CPXCR1 has revealed several sequence variants present on all affected CPX chromosomes. However, these variants have also been detected at a lower frequency on unaffected chromosomes, indicating that they are polymorphisms that are unlikely to cause the CPX phenotype. PMID- 11499682 TI - Analysis of lymphoedema-distichiasis families for FOXC2 mutations reveals small insertions and deletions throughout the gene. AB - Lymphoedema-distichiasis (LD) is a dominantly inherited form of primary lymphoedema with onset of lower limb swelling at puberty or later. There is variable penetrance of this disorder, but the most consistently inherited feature is distichiasis, viz. fine hairs arising inappropriately from the meibomian glands. We established linkage of this disorder to 16q24.3 and the gene has recently been identified as the forkhead transcription factor FOXC2. We report the mutational analysis of 14 families with LD. All but one of these pedigrees have small insertions or deletions in the gene, which seem likely to produce haploinsufficiency. The mutation sites are scattered throughout the gene. There is one family with a mis-sense mutation in the forkhead domain of the protein. This base alteration is not a common polymorphism, is co-inherited with the disease and produces a non-conservative amino acid change. PMID- 11499683 TI - The TNFRSF6 gene is not implicated in familial early-onset Alzheimer's disease. PMID- 11499684 TI - Doxorubicin/paclitaxel combination does not expose breast cancer patients to excessive cardiac risk. PMID- 11499685 TI - Atrasentan prolongs time to progression in prostate cancer patients. PMID- 11499686 TI - Excessive paperwork detracts from patient care, professional mentoring, and research. PMID- 11499687 TI - Prostate cancer and spinal cord compression. AB - Prostate cancer metastasis to the spine is an extremely difficult clinical problem to treat. However, it occurs commonly, and all clinicians--not only oncologists--should undertake to understand its pathogenesis, diagnosis, clinical presentation, and current treatment options. This review emphasizes the surgical treatment of prostate cancer metastasis to the spine. The goals of this article are to (1) present an overview of the pathophysiology of this disease, with an emphasis on the mechanisms of metastasis and invasion, (2) provide a general overview of the clinical presentation and diagnosis of metastatic prostate carcinoma, and (3) discuss currently available treatment options. Such options include best medical management, nonsurgical treatments (radiation, chemotherapy), and surgical treatment of newly diagnosed and previously irradiated metastatic prostate carcinoma to the spine. Algorithms for the treatment of this disease are presented. PMID- 11499688 TI - Current status of thalidomide in the treatment of cancer. AB - Tumor angiogenesis is a critical factor in the growth and metastasis of most malignant neoplasms. Thalidomide (Thalomid), banned from clinical use in the 1960s because of severe teratogenicity, has been shown to possess antiangiogenic properties. A recent clinical trial of antiangiogenic therapy with thalidomide demonstrated significant activity in a group of patients with relapsed refractory myeloma. Although its mechanism of action remains unclear, several trials have since confirmed that thalidomide is active in 25% to 35% of patients with relapsed myeloma. As a result, thalidomide has reemerged in clinical practice and is now actively being studied in the treatment of several cancers. Major toxicities associated with the use of thalidomide include constipation, sedation, skin rash, fatigue, and peripheral neuropathy. This article summarizes the current status of thalidomide therapy in cancer. PMID- 11499689 TI - Clinical trials referral resource. ST1571. PMID- 11499690 TI - Risk of breast and ovarian cancer in women with strong family histories. AB - Assessing the risk of breast and ovarian cancer starts with obtaining a complete and accurate family history. This can reveal evidence of inherited cancer risk. The highest risk of cancer is associated with germ-line abnormalities in several genes, including BRCA1, BRCA2, and TP53. Moderate-risk genes associated with syndromes that are inherited in an autosomal dominant pattern (such as Cowden's disease, hereditary non-polyposis colorectal cancer, Muir-Torre syndrome, and Peutz-Jeghers syndrome) exhibit lower penetrance and thus less risk of breast and/or ovarian cancer. Low-risk genes likely require significant environmental exposure, and although they are associated with the lowest risk of cancer, they account for more cancer than high- and moderate-risk genes. Lifetime risks for breast or ovarian cancer can be estimated. The Gail and Claus models, the more widely utilized models for calculation of lifetime breast cancer risk, are discussed. Models are also available for determining the likelihood of finding a BRCA1/2 mutation (the BRCAPRO and Myriad models). Appropriate candidates for testing include affected individuals who are most likely to have a hereditary form of cancer. Testing should proceed only after a thorough discussion of the risks, benefits, and limitations of testing. Risk-reducing options are available to women with a strong family history of breast and ovarian cancer. These options include high-risk screening, chemoprevention, and prophylactic surgery. PMID- 11499691 TI - Gemcitabine active in patients with metastatic breast cancer. PMID- 11499692 TI - Docetaxel/carboplatin combination produces impressive response in ovarian cancer patients. PMID- 11499693 TI - Pancreatic cancer in the older patient. AB - Pancreatic cancer is a disease seen predominantly in elderly patients. Compared to younger patients, older patients are more likely to present with early-stage disease and, therefore, may be candidates for aggressive local treatment. Little published information exists on treatment outcomes for elderly patients with potentially resectable disease or those with locally advanced or metastatic pancreatic cancer. The limited information available suggests that elderly patients are as likely to benefit from surgery, radiation, and chemotherapy as younger patients. Despite this apparent benefit, elderly patients appear to have a worse long-term outcome. This may be due to the failure to offer them aggressive treatment or to comorbid conditions. Nevertheless, further studies need to be conducted in this area, and greater emphasis needs to be placed on including elderly patients in clinical trials. For elderly patients with terminal disease, there should be better use of palliative measures that may be of benefit. Each of these issues is discussed in detail. PMID- 11499694 TI - Use of in vitro assays to assess hematotoxic effects of environmental compounds. AB - The number of chemicals being introduced into the environment increases and many of these substances may pose a health risk to exposed individuals. Many environmental toxicants with a potential toxicity to the hematopoietic system have been identified by animal experiments. Owing to the risks of severe chronic hematopoietic disorders, it is important to screen chemicals for their hematotoxicity. The aim of this work was to identify, through the use of in vitro techniques, targets for hematotoxic effects. Our study focused on myeloid and erythroid hematopoietic progenitors and stromal stem cells as possible targets. The in vitro assays showed that various hematotoxic compounds exert different effects on these cell populations. In vitro exposure of murine bone marrow cells to various inorganic (cadmium, lead) and organic (benzene metabolites, lindane. benzo-[a]-pyrene (BaP), PCB (polychlorinated biphenyl) congeners) environmental chemicals indicated that hematopoietic or stromal bone marrow cells were targets for most of the chemicals. Stromal cells were more affected by lead, cadmium, and BaP compared to myeloid cells. Benzene and phenol gave no response, but the metabolites catechol and hydroquinone were equally toxic to the stromal and the myeloid progenitor cells. Among the PCBs tested, PCB126 was most toxic. Human progenitor cells derived from cord blood were exposed in vitro to catechol, hydroquinone, lead nitrate, and PCBs. Human hematopoietic cells were sensitive to the tested compounds. Human erythroid progenitors are more susceptible to lead exposure than are myeloid progenitors. Based on the in vitro tests, humans are more sensitive to lead, catechol, and PCB126 than are mice. In contrast to the murine data, humans responded with individual differences to lead and PCB126. PMID- 11499695 TI - Antioxidant activity of resveratrol in endotoxin-stimulated blood platelets. AB - Resveratrol (3,4',5-trihydroxystilbene) is a natural molecule with antioxidant action. It is also considered to be a molecule with antiplatelet, anticancer and anti-inflammatory action. The effects of trans-resveratrol on the reactive oxygen species (ROS) generation and thiobarbituric acid-reactive substances (TBARS) in blood platelets induced by endotoxin (lipopolysaccharide, LPS) or thrombin were studied in vitro. The production of superoxide radicals (O2.-) and other reactive oxygen species (H2O2, singlet oxygen, and organic radicals) in the presence of resveratrol was measured by a chemiluminescence method in resting blood platelets and platelets stimulated by LPS (0.3 microg/ 10(8) platelets) or thrombin (2.5 U/10(8) platelets). We have shown that resveratrol (6.25-100 microg/ml) inhibits chemiluminescence and generation of O2.- in blood platelets. It has an inhibitory effect on the production of ROS and TBARS in platelets caused by LPS or thrombin. PMID- 11499696 TI - Comparative study of the cytotoxicity and apoptosis-inducing potential of commonly occurring oxysterols. AB - The cytotoxicity of the oxysterols 25-hydroxycholesterol, 7beta hydroxycholesterol, cholesterol-5alpha,6alphaepoxide, cholesterol-5beta,6beta epoxide, 19-hydroxycholesterol and 7-ketocholesterol was examined in U937 cells, a human monocytic blood cell line. 7beta-Hydroxycholesterol, cholesterol 5beta,6beta-epoxide, and 7-ketocholesterol, at 30 micromol/L concentration, were found to be cytotoxic to this cell line and the mode of cell death was by apoptosis. 25-Hydroxycholesterol, cholesterol-5alpha,6beta-epoxide and 19 hydroxycholesterol (30 micromol/L) did not induce apoptosis in this cell line. Since it has been suggested that the generation of an oxidative stress may occur in the early stages of the apoptotic process, the glutathione concentration and the activity of superoxide dismutase were also measured in the oxysterol-treated cells. 7beta-Hydroxycholesterol was shown to increase the superoxide dismutase activity and decrease the glutathione concentration. However, cholesterol 5beta,6beta-epoxide and 7-ketocholesterol, which were also shown to induce apoptosis, did not affect the glutathione concentration or the superoxide dismutase activity in the U937 cells. Therefore, oxysterol-induced apoptosis may not be dependent on the generation of an oxidative stress. PMID- 11499697 TI - Cancer vaccines based on dendritic cells loaded with tumor-associated antigens. AB - Dendritic cells (DCs) can be used as an antigen presentation platform for vaccination against cancer. In this approach, DCs are expanded in vitro from monocyte-derived progenitors, and subsequently loaded with well-characterized tumor-associated antigens (TAAs). TAAs can be incubated with DCs in various forms, including peptides, recombinant proteins, plasmid DNA, formulated RNA, or recombinant viruses. Advantages and limitations of DC-based cellular vaccines against cancers, as well as preliminary results of clinical studies already performed in humans, are discussed. Importantly, significant advances in our understanding of the biology of DCs can be used to support the design of new vaccines or adjuvants in order to elicit TH1 cellular immune responses. PMID- 11499698 TI - Liver transplantation of hepatic stem cells: potential use for treating liver diseases. AB - Hepatic stem cells are an alternative means for repopulating the liver after various injuries instead of liver transplantation. The first step before use is to select stem cells that will be good candidates. This review discusses the different candidates including fetal progenitor bipotential hepatic stem cells; adult hepatocytes, which can be considered as unipotential committed stem cells; and oval cells, a type of nonparenchymal pluripotential hepatic stem cell. The advantages and disadvantages of each type of cell are discussed and several other possible alternatives, such as the use of hematopoietic stem cells are analyzed. PMID- 11499699 TI - Relevance of clonogenic assays in hematotoxicology. AB - Clonogenic assays have been established in hematology for 30 years. They have been widely used in fundamental studies on hematopoiesis and they are also routinely used in clinical hematology to confirm diagnosis or to predict time to recovery in cases of bone marrow failure. Their use in toxicological studies is more recent. Adverse effects of xenobiotics can induce hematological problems and pathologies such as neutropenia, thrombocytopenia, anemia, and aplastic anemia. Three clonogenic assays are proposed for granulopoiesis, megakaryopoieisis and erythropoieisis. Hematopoietic progenitors from murine or human origin can be cultured in the presence of xenobiotics using validated protocols to complete standard animal toxicological studies. These clonogenic assays can help to predict adverse effects of drugs or toxicants. Clonogenic assays using white blood cell progenitors (CFU-GM culture) have recently been validated by ECVAM and can be used routinely. Megakaryocyte progenitor (CFU-MK) culture is under development and prevalidation in toxicological studies supported by ECVAM. Red blood cells progenitor culture (BFU-E) has been proposed but needs international validation to be recognized. PMID- 11499700 TI - In vitro bone marrow granulocyte-macrophage progenitor cultures in the assessment of hematotoxic potential of the new drugs. AB - In pharmaceutical research, in vitro toxicity tests, for assessing the potential toxicity of new chemical entities are necessary in the early stages of the developmental process, when no information is available about the metabolism or even the target organ toxicity of the compounds to be tested. In vitro specific organ toxicity tests, such as the granulocyte-macrophage colony-forming unit (CFU GM) clonogenic assay, are useful tools for predicting the adverse effects of new compounds on the blood-forming system, provided that some reference points are available, e.g., toxicological information about compounds belonging to the same chemical class and structure-activity relationship data. Furthermore, when no information is available about metabolism, the in vitro system should cover as many possibilities as possible, to avoid false positive or false negative results. In fact, while many compounds are metabolized to a variety of inactive chemical species, some undergo bioactivation to form more active metabolites. The addition of a metabolic activation system to the CFU-GM assay enables assessment of direct and metabolism-mediated toxicity. The regulatory agencies and industry value the concept of assays performed with and without metabolic activation, since they often have to take decisions about compounds with unknown mechanisms of action. CFU-GM assay, designed in this way, is an example of such a mechanism naive assay. It has been suggested that, for new compounds, metabolites should be generated and tested both in the presence and in the absence of the parent compound itself, to identify the possible contribution of metabolites to the hematotoxicity observed, and to determine whether there is any synergistic or antagonistic effect between metabolites and the parent compound that might affect hematotoxicity in vivo. Various approaches can be used to obtain such information. PMID- 11499701 TI - Magnetic resonance imaging of spongy degeneration of the central nervous system in a Labrador Retriever. AB - A 7-month-old, neutered female Labrador Retriever was evaluated for tetraparesis and subtle cerebellar dysfunction. Clinical signs progressed over a period of 6 weeks to severe ataxia, hypermetria, intention tremors, and finally non ambulatory tetraparesis. On magnetic resonance imaging of the brain there were large, bilaterally symmetrical, ovoid lesions in the region of the deep cerebellar nuclei that were hyperintense on T2-weighted and proton density images and hypointense on T1-weighted images. There were similar but smaller bilaterally symmetrical lesions present within the thalamus. Euthanasia was performed and lesions consistent with the previously described spongy degeneration of Labrador Retrievers were identified. This disease and its relation to similar human heritable leukodystrophies are discussed. PMID- 11499702 TI - Magnetic resonance imaging features of equine nigropallidal encephalomalacia. AB - Magnetic resonance imaging (MR) was used to make a diagnosis of equine nigropallidal encephalomalacia in a horse. Equine nigropallidal encephalomalacia is a neurodegenerative disease that has many characteristics with Parkinson-like diseases in humans. Historically, horses were euthanized based on clinical signs and exposure to the toxic weed, yellow star thistle (Centaurea solstitialis). Previously, the disease has only been confirmed on necropsy. MR imaging can provide accurate and sensitive visualization of typical lesions seen in the brain of horses affected with equine nigropallidal encephalomalacia. Lesions were seen on T1-weighted, T2-weighted and proton density images. There was no contrast enhancement following Gd-DTPA administration. Lesions seen on MR were confirmed at necropsy. Using MR to confirm a diagnosis of equine nigropallidal encephalomalacia will prevent unnecessary suffering of horses and expense to owners that would otherwise incur, while further diagnostics are performed. PMID- 11499703 TI - The canine trochanteric fossa: a radiographic and anatomic study. AB - Radiographic features of the region of the trochanteric fossa were evaluated bilaterally on 175 ventrodorsal radiographs of canine hip joints and findings were compared to anatomic specimens. The fossa showed a variable radiographic appearance, which was affected by positioning and anatomic diversity. On a small number of radiographs (n = 12), the trochanteric fossa was clearly defined as a circular area of radiopacity surrounded by a radiolucent halo. This appearance was significantly influenced by medial rotation of the femur (P < 0.001). On some anatomic specimens an irregular protuberance representing the site of attachment of the gemelli, internal and external obturator muscles was seen within the fossa, surrounded by a variable number of small openings, which were thought to be nutrient foramina. Dissection identified a small number of minor branches of the medial circumflex femoral artery entering these. PMID- 11499704 TI - Radiographic findings in the thorax of dogs with leptospiral infection. AB - Thoracic radiographs of 4 dogs with confirmed and 1 dog with suspected leptospirosis were reviewed. In all dogs a reticulonodular pulmonary opacity was noted, affecting the entire lung in 3 and predominantly the caudodorsal lung field in 2 dogs. The radiographic lung pattern described is associated with pulmonary hemorrhage probably due to endothelial damage and vasculitis. Pulmonary manifestations in dogs with leptospirosis may be misinterpreted and attributed to neoplasia, pneumonia, edema, hemorrhage due to disseminated intravascular coagulopathy, adult respiratory distress syndrome and pulmonary thromboembolism. PMID- 11499705 TI - Radiographic diagnosis: pericardioperitoneal diaphragmatic hernia and cholelithiasis in a dog. PMID- 11499706 TI - An integrated epidemiologic and radiographic algorithm for canine urocystolith mineral type prediction. AB - Research involved 2 databases. One database (occurrence frequency) comprised the age, breed, gender and urocystolith mineral type (pure chemical types only) from 2041 canine patients submitted to the Minnesota Urolith Center. The other database (imaging) comprised the maximum size, surface (rough, smooth, and smooth with blunt tips), shape (faceted, irregular, jackstone, ovoid, and round) and internal architecture (lucent center, random-nonuniform, and uniform) from 434 canine patients imaged in a urinary bladder phantom. The imaging database was a partial subset of the occurrence frequency database. Imaging techniques simulated were survey radiography and double contrast cystography. The databases were compared using multivariate analysis techniques. Equations were developed to use clinically-relevant characteristics (age, breed, gender, maximum size, surface, shape, and internal architecture) to predict urocystolith mineral types. The goal was to assess the accuracy of the various techniques in predicting the urocystolith mineral types. The combination of signalment (age, breed, gender) and simulated survey radiographic findings does not improve mineral type prediction accuracy (average across all mineral types is 69.9%) beyond that achievable with signalment alone (average across all mineral types is 69.8%). However, the combination of signalment and double contrast cystography does improve mineral type prediction accuracy (average across all mineral types is 75.3%). For comparison, mineral type prediction accuracy without signalment from survey radiographs only was 65.7% across all mineral types. The clinical utility of the algorithm is the option to distinguish urocystolith mineral types requiring surgical vs. medical treatment. PMID- 11499707 TI - Anatomic models and phantoms for diagnostic ultrasound instruction. AB - The preparation and application of anatomic models and phantoms to facilitate learning diagnostic ultrasound is described. Imaging with diagnostic ultrasound requires mastery of many skills, along with knowledge of sound-tissue interactions which contribute to the formation of diagnostic images and artifacts. Understanding the genesis of artifacts encountered during ultrasound scanning can avoid misinterpretation and aid diagnosis. In addition, development of machine related knowledge and skills, including manipulation of the transducer and the selection of correct settings for variables such as gain, power, time gain compensation, and transducer type, is dependent on an understanding of how these factors affect the image. The normal appearance of an organ relates to both its echogenicity and morphologic characteristics, and confirmation of the nature of an abnormality often requires ultrasound guided biopsy. The use of anatomic models and phantoms in ultrasound instruction allows principles to be demonstrated, knowledge acquired, and biopsy procedures practiced and mastered in a controlled setting. This can minimize live animal use, and enhance the knowledge base and skills of the clinician prior to applying this diagnostic technique to the clinical patient. PMID- 11499708 TI - Ultrasonography of the normal feline pancreas and associated anatomic landmarks: a prospective study of 20 cats. AB - The sonographic appearance of the feline pancreas and associated anatomic landmarks including the pancreatic duct, duodenum, duodenal papilla, portal vein, and gastric lymph node were evaluated in 20 healthy, awake cats. The pancreas appeared nearly isoechoic to surrounding mesenteric tissues, isoechoic to slightly hyperechoic to adjacent liver lobes, and hypoechoic to the spleen. The mean thickness measurements for the right pancreatic lobe, body, and left pancreatic lobe were 4.5 mm (range 2.8-5.9), 6.6 mm (range 4.7-9.5), and 5.4 mm (range 3.4-9.0), respectively. The pancreatic duct was consistently visualized in the left pancreatic lobe and had a mean thickness of 0.8 mm (range 0.5-1.3). It could be differentiated from the pancreatic vessel, by its central location, and the duct's lack of Doppler flow signal. The duodenum was used as a landmark to identify the right lobe of the pancreas. The mean duodenal wall thickness measurement was 2.8 mm (range 2.1-3.8) in sagittal section, and 3.0 mm (range 2.2 4.4) in transverse section. The duodenal papilla was identified in 4 of 20 cats. It ranged in size from 2.9 to 5.5 mm in width, and had a maximum height of 4.0 mm in transverse section. The portal vein was used as a consistent anatomic landmark for identification of the left lobe and body of the pancreas. The mean diameter of the portal vein at the level where the pancreatic body joins the left pancreatic lobe was 4.3 mm (range 2.7-5.9) when viewed in sagittal section, and 4.5 mm (range 3.6-6.1) in transverse section. The gastric lymph node was identified cranial and ventromedial to the pyloroduodenal angle in 6 of 20 cats. It had an asymmetrical shape with a larger caudal pole in five of the six cats. The largest dimensions of the gastric lymph node were 10 mm in length, and 6 mm in width for the larger caudal pole, and 5.1 mm in width for the smaller cranial pole. PMID- 11499709 TI - The role of ultrasound in the assessment of laryngeal paralysis in the dog. AB - Forty dogs with clinical signs suggestive of upper respiratory tract disease underwent echolaryngography and laryngoscopy. Laryngoscopy was used as the definitive technique to diagnose laryngeal paralysis. The ultrasound investigation accurately indicated the presence of the paralysis and confirmed the uni- or bilateral nature of the disorder. Findings indicative of laryngeal paralysis included asymmetry or absence of motion of the cuneiform processes (30/30), abnormal arytenoid movement (16/30), paradoxical movement (9/30), caudal displacement of the larynx (2/30) and laryngeal collapse (1/30). Thirty dogs were found to be afflicted with laryngeal paralysis and ten had normal laryngeal motility. PMID- 11499710 TI - Radiographic and ultrasonographic findings of liver lobe torsion in a dog. AB - A twelve-year-old neutered male beagle presented for a cranial abdominal mass. The results of physical examination, laboratory tests, radiography, and ultrasonography are presented. A torsion of the quadrate lobe of the liver was diagnosed at surgery and confirmed by histopathology. A brief discussion of liver lobe torsion is presented. PMID- 11499711 TI - Canine splenic myelolipomas. PMID- 11499712 TI - Percutaneous ultrasound-guided pyelography aided diagnosis of ectopic ureter and hydronephrosis in a 3-week-old filly. AB - Ultrasound-guided percutaneous pyelography has previously been used in dogs to diagnose ectopic ureter. To the authors' knowledge its use has never been reported in the horse. This paper reports the technique used in a three-week-old filly with a suspected ectopic ureter. Pyelography can be performed under sedation and is a relatively simple, safe procedure. The technique enables assessment of individual renal function through collection and analysis of urine from each renal pelvis. PMID- 11499713 TI - Ultrasonographic diagnosis of ovine cystic echinococcosis. AB - The sanitary and economic impact of cystic echinococcosis is serious in those countries where it becomes endemic. Ultrasonography is one technique that may be used to diagnose this disease in endemic areas. In parasitized sheep, hydatid cysts appear sonographically as a round hypoechoic structure. Twenty two sheep destined for slaughter were studied sonographically and imaging findings compared to post-mortem findings. Three sheep with hydatid cysts were identified. Eighty additional sheep not destined for slaughter were also studied. Echinococcus granulosus cysts were detected in three animals. Forty sheep from a non-endemic area had no hepatic cysts. The in vivo sonographic study of sheep provides a useful screening tool for echinococcosis. PMID- 11499714 TI - Ultrasound appearance of an equine testicular seminoma. AB - A 16-year-old thoroughbred stallion developed sudden swelling of the left testicle. The stallion had previously been regarded as a unilateral cryptorchid. Ultrasound examination of the left testicle revealed a diffusely heterogeneous parenchyma. The testicle was diffusely hypoechoic with ill defined regions of hyperechogenicity giving the appearance of hypoechoic nodules throughout the testicular parenchyma. No normal testicular tissue was identifiable. An echogenic band, representing a pseudocapsule could be seen surrounding the testicle. Histopathologic diagnosis a seminoma. PMID- 11499715 TI - A retrospective study into the effects of operator experience on the accuracy of ultrasound in the diagnosis of gastric neoplasia in dogs. PMID- 11499716 TI - Arterial remodeling and coronary artery disease: the concept of "dilated" versus "obstructive" coronary atherosclerosis. AB - Traditionally, the development of coronary artery disease (CAD) was described as a gradual growth of plaques within the intima of the vessel. The outer boundaries of the intima, the media and the external elastic membrane (EEM), were thought to be fixed in size. In this model plaque growth would always lead to luminal narrowing and the number and severity of angiographic stenoses would reflect the extent of coronary disease. However, histologic studies demonstrated that certain plaques do not reduce luminal size, presumably because of expansion of the media and EEM during atheroma development. This phenomenon of "arterial remodeling" was confirmed in necropsy specimens of human coronary arteries. More recently, the development of contemporary imaging technology, particularly intravascular ultrasound, has allowed the study of arterial remodeling in vivo. These new imaging modalities have confirmed that plaque progression and regression are not closely related to luminal size. In this review, we will analyze the role of remodeling in the progression and regression of native CAD, as well as its impact on restenosis after coronary intervention. PMID- 11499717 TI - Nuclear factor kappa-B and the heart. AB - Nuclear factor kappa-B (NFkappaB), a redox-sensitive transcription factor regulating a battery of inflammatory genes, has been indicated to play a role in the development of numerous pathological states. Activation of NFkappaB induces gene programs leading to transcription of factors that promote inflammation, such as leukocyte adhesion molecules, cytokines, and chemokines, although some few substances with possible anti-inflammatory effects are also NFkappaB regulated. The present article reviews basic regulation of NFkappaB and its activation, cell biological effects of NFkappaB activation and the role of NFkappaB in apoptosis. Evidence involving NFkappaB as a key factor in the pathophysiology of ischemia reperfusion injury and heart failure is discussed. Although activation of NFkappaB induces pro-inflammatory genes, it has lately been indicated that the transcription factor is involved in the signaling of endogenous myocardial protection evoked by ischemic preconditioning. A possible role of NFkappaB in the development of atherosclerosis and unstable coronary syndromes is discussed. Nuclear factor kappa-B may be a new therapeutic target for myocardial protection. PMID- 11499718 TI - Development of left ventricular hypertrophy in adults in hypertrophic cardiomyopathy caused by cardiac myosin-binding protein C gene mutations. AB - OBJECTIVES: We sought to determine whether the development of left ventricular hypertrophy (LVH) can be demonstrated during adulthood in genetically affected relatives with hypertrophic cardiomyopathy (HCM). BACKGROUND: Hypertrophic cardiomyopathy is a heterogeneous cardiac disease caused by mutations in nine genes that encode proteins of the sarcomere. Mutations in cardiac myosin-binding protein C (MyBPC) gene have been associated with age-related penetrance. METHODS: To further analyze dormancy of LVH in patients with HCM, we studied, using echocardiography and 12-lead electrocardiography, the phenotypic expression caused by MyBPC mutations in seven genotyped pedigrees. RESULTS: Of 119 family members studied, 61 were identified with a MyBPC mutation, including 21 genetically affected relatives (34%) who did not express the HCM morphologic phenotype (by virtue of showing normal left ventricular wall thickness). Of these 21 phenotype-negative individuals, 9 were children, presumably in the prehypertrophic phase, and 12 were adults. Of the 12 adults with normal wall thickness < or = 12 mm (7 also with normal electrocardiograms), 5 subsequently underwent serial echocardiography prospectively over four to six years. Of note, three of these five adults showed development of LVH in mid-life, appearing for the first time at 33, 34 and 42 years of age, respectively, not associated with outflow obstruction or significant symptoms. CONCLUSIONS: In adults with HCM, disease-causing MyBPC mutations are not uncommonly associated with absence of LVH on echocardiogram. Delayed remodeling with the development of LVH appearing de novo in adulthood, demonstrated here for the first time in individual patients with prospectively obtained serial echocardiograms, substantiates the principle of age-related penetrance for MyBPC mutations in HCM. These observations alter prevailing perceptions regarding the HCM clinical spectrum and family screening strategies and further characterize the evolution of LVH in this disease. PMID- 11499719 TI - Spectrum of clinical phenotypes and gene variants in cardiac myosin-binding protein C mutation carriers with hypertrophic cardiomyopathy. AB - OBJECTIVES: We studied the clinical and genetic features of hypertrophic cardiomyopathy (HCM) caused by mutations in the myosin-binding protein C gene (MYBPC3) in 110 consecutive, unrelated patients and family members of European descent. BACKGROUND: Mutations in the MYBPC3 gene represent the cause of HCM in approximately 15% of familial cases. MYBPC3 mutations were reported to include mainly nonsense versus missense mutations and to be characterized by a delayed onset and benign clinical course of the disease in Japanese and French families. We investigated the features that characterize MYBPC3 variants in a large, unrelated cohort of consecutive patients. METHODS: The MYBPC3 gene was screened by single-strand conformational polymorphism analysis and sequencing. The clinical phenotypes were analyzed using rest and 24-h electrocardiography, electrophysiology, two-dimensional and Doppler echocardiography and angiography. RESULTS: We identified 13 mutations in the MYBPC3 gene: one nonsense, four missense and three splicing mutations and five small deletions and insertions. Of these, 11 were novel, and two were probably founder mutations. Patients with MYBPC3 mutations presented a broad range of phenotypes. In general, the 16 carriers of protein truncations had a tendency toward earlier disease manifestations (33 +/- 13 vs. 48 +/- 9 years; p = 0.06) and more frequently needed invasive procedures (septal ablation or cardioverter-defibrillator implantation) compared with the 9 carriers of missense mutations or in-frame deletions (12/16 vs. 1/9 patients; p < 0.01). CONCLUSIONS: Multiple mutations, which include missense, nonsense and splicing mutations, as well as small deletions and insertions, occur in the MYBPC3 gene. Protein truncation mutations seem to cause a more severe disease phenotype than missense mutations or in-frame deletions. PMID- 11499720 TI - On genetic and phenotypic variability of hypertrophic cardiomyopathy: nature versus nurture. PMID- 11499721 TI - Biphasic changes in left ventricular end-diastolic pressure during dynamic exercise in patients with nonobstructive hypertrophic cardiomyopathy. AB - OBJECTIVES: The aim of this study was to clarify the serial changes in left ventricular (LV) end-diastolic pressure (LVEDP) during dynamic exercise in patients with hypertrophic cardiomyopathy (HCM). BACKGROUND: Although HCM is characterized by impaired resting LV diastolic function, serial changes in LVEDP during exercise have not been characterized. METHODS: We simultaneously measured LV pressure and LV dimensions during symptom-limited supine bicycle exercise in 5 healthy individuals and 20 patients with HCM. Exercise thallium-201 scintigraphic studies were also performed. RESULTS: The LVEDP (baseline: 12 +/- 5 mm Hg) progressively increased to a maximum value at peak exercise (28 +/- 8 mm Hg) in 11 patients with HCM (group I). In the remaining nine patients with HCM (group II), changes in LVEDP during exercise were biphasic, with an initial progressive increase and a subsequent gradual decline up to peak exercise (14 +/- 4 mm Hg at baseline, 27 +/- 5 mm Hg at the critical heart rate, 16 +/- 3 mm Hg at peak exercise). Exercise-induced changes in LV dimensions and LV peak systolic pressures were similar in both groups. However, the maximum first derivative of LV pressure was greater and the LV pressure half-time was shorter in group II than in group I at a similar peak exercise heart rate. The biphasic changes in LVEDP disappeared by pretreatment with propranolol. The LV hypertrophy scores were higher in group I than in group II. Exercise thallium-201 images showed more severe perfusion defects in group I than in group II patients. CONCLUSIONS: The biphasic changes in LVEDP seen during exercise may be related to improved coronary microcirculation in response to beta-adrenergic stimulation in patients with mild to moderate HCM. PMID- 11499722 TI - Antiarrhythmic drug therapy in the Multicenter UnSustained Tachycardia Trial (MUSTT): drug testing and as-treated analysis. AB - OBJECTIVES: Using data from the Multicenter UnSustained Tachycardia Trial (MUSTT), we examined the factors used to select antiarrhythmic drug therapy and their impact on outcomes. BACKGROUND: The MUSTT examined the use of programmed ventricular stimulation (PVS) to guide antiarrhythmic therapy in patients with coronary arteriosclerosis, left ventricular dysfunction and asymptomatic, unsustained ventricular tachycardia (VT). Trial outcomes may reflect factors used to select antiarrhythmic drug therapy. METHODS: We compared subgroups of patients with inducible sustained VT randomized to PVS-guided antiarrhythmic therapy (n = 351), in particular those receiving PVS-guided antiarrhythmic drug therapy (n = 142) versus no antiarrhythmic therapy (controls, n = 353). RESULTS: "Effective" antiarrhythmic drug therapy (i.e., the term "effective" was used to denote therapy that resulted in noninducible VT or hemodynamically stable induced VT) was found for 142 of the 351 patients (43%), most often at the first or second PVS session (125/142, 88%). Mortality among the 142 patients did not differ from that among control patients. Of these 142 patients, the PVS end point was noninducibility in 91 patients and stable VT in 51 patients. Mortality did not differ between these two groups either, but arrhythmia was numerically more frequent in the PVS-induced stable VT group. Mortality was greatest in the few patients receiving propafenone (unadjusted p = 0.07, adjusted p = 0.14 vs. controls), but mortality with all agents did not differ from that of controls, even after adjustment. CONCLUSIONS: Even when presenting the results as favorably as possible, we found no benefit with PVS-guided drug therapy in patients with clinical unsustained VT who had inducible sustained VT. These findings are unaltered by using different end points for PVS or considering the response to individual drugs. PMID- 11499723 TI - Mindfully mining MUSTT. Multicenter Unsustained Tachycardia Trial. PMID- 11499724 TI - Prevalence, characteristics and clinical implications of regular atrial tachyarrhythmias in patients with atrial fibrillation: insights from a study using a new implantable device. AB - OBJECTIVES: This study prospectively analyzed atrial tachyarrhythmia (AT) organization and antitachycardia pacing (ATP) success in patients with an implanted device for AT therapy. BACKGROUND: In patients with atrial fibrillation (AF), the incidence of regular, slow ATs amendable by ATP is unknown. METHODS: Forty patients with previously documented AT (70% with AF) received a new pacemaker with atrial electrogram (AEG) storage and atrial ATP capabilities for standard pacing indications. The AEGs acquired during the first month (study phase 1) were classified into high (type I), intermediate (type II) and low (type III) degrees of organization. Atrial ATP was then activated, and treated AT episodes were retrieved three and six months after implantation (study phase 2). RESULTS: Of 824 AEGs retrieved before ATP activation (study phase 1), 351 (43%) were classified as type 1, 47% as type II and 10% as type III. Episodes of AT starting as type I (35%) and type II or III (65%) maintained their type over 1 min in 73%. All patients with an exclusive history of AF also showed type I AEGs. In 361 subsequently treated AT episodes (study phase 2), ATP was successful in 62% of type I and 34% of type II episodes, but not in type III (p < 0.0001). CONCLUSIONS: The majority of patients with a history of AF show not only disorganized but also highly organized AT episodes, which can be successfully terminated by ATP. PMID- 11499725 TI - Long-term follow-up of right ventricular monomorphic extrasystoles. AB - OBJECTIVES: The purpose of this study was to verify in a long-term follow-up whether frequent monomorphic right ventricle extrasystoles may progress to arrhythmogenic right ventricular dysplasia (ARVD). BACKGROUND: Frequent monomorphic right ventricle extrasystoles are generally considered benign. However, in patients with this pattern, cardiac magnetic resonance (MR) has recently shown anatomical and functional abnormalities of the right ventricle. METHODS: Sixty-one patients who had been classified by noninvasive examinations as having frequent idiopathic right ventricle ectopy were contacted after 15 +/- 2 years (12 to 20) and submitted to clinical examination, electrocardiogram (ECG), Holter monitoring, stress test, signal averaged ECG, echocardiography and, in 11 patients, cardiac MR. The primary end point was to ascertain the presence of cases of sudden death or progression to ARVD. RESULTS: At the end of the follow-up, 55 patients were alive; six died, none of sudden death; eight stated to be well but refused further examinations. The 47 patients examined had normal ECG; in 24 patients (51%), extrasystoles were no longer present at Holter monitoring; late potentials were present in up to 15% of the patients; the right ventricle was normal at echocardiography. In 8 of 11 patients (73%), cardiac MR showed focal fatty replacement and other abnormalities of the right ventricle. CONCLUSIONS: In this long-term follow-up study, no patient died of sudden death nor developed ARVD; two-thirds of the patients were asymptomatic, and, in half of the patients, ectopy had disappeared. Focal fatty replacement in the right ventricle was present in most. PMID- 11499727 TI - Coexistence of type I atrial flutter and intra-atrial re-entrant tachycardia in patients with surgically corrected congenital heart disease. AB - OBJECTIVES: This study assessed the coexistence of intra-atrial re-entrant tachycardia (IART) and isthmus-dependent atrial flutter (IDAF) in patients presenting with supraventricular tachyarrhythmias after surgical correction of congenital heart disease (CHD). BACKGROUND: In patients with CHD, atrial tachyarrhythmias may result from IART or IDAF. The frequency with which IART and IDAF coexist is not well defined. METHODS: Both IDAF and IART were diagnosed in 16 consecutive patients using standard criteria and entrainment mapping. Seven patients had classic atrial flutter morphology on surface electrocardiogram (ECG), whereas nine had atypical morphology. RESULTS: A total of 24 circuits were identified. Three patients had IDAF only, five had IART only, seven had both, and one had a low right atrial wall tachycardia that could not be entrained. Twenty two different reentry circuits were ablated. Successful ablation was accomplished in 13 of 14 (93%) IART and 9 of 10 (90%) IDAF circuits. There was one IART recurrence. The slow conduction zone involved the region of the right atriotomy scar in 12 of 14 (86%) IART circuits. No procedural complications and no further recurrences were seen after a mean follow-up of 24 months. CONCLUSIONS: Both IDAF and IART are the most common mechanisms of atrial re-entrant tachyarrhythmias in patients with surgically corrected CHD, and they frequently coexist. The surface ECG is a poor tool for identifying patients with coexistent arrhythmias. The majority of IART circuits involve the lateral right atrium and may be successfully ablated by creating a lesion extending to the inferior vena cava. PMID- 11499726 TI - Significance of inducible ventricular fibrillation in patients with coronary artery disease and unexplained syncope. AB - OBJECTIVES: This study was designed to determine the incidence and prognostic significance of inducible ventricular fibrillation (VF) in patients with coronary artery disease (CAD) and unexplained syncope. BACKGROUND: Current American College of Cardiology/American Heart Association practice guidelines recommend implantation of internal cardioverter-defibrillators (ICDs) in patients with unexplained syncope in whom either ventricular tachycardia (VT) or VF is inducible during electrophysiologic (EP) testing. Although the prognostic significance of inducible monomorphic VT is known, the significance of inducible VF remains undefined. METHODS: We evaluated 118 consecutive patients with CAD and unexplained syncope who underwent EP testing. Sustained monomorphic VT was inducible in 53 (45%) patients; in 20 (17%) patients, VF was the only inducible arrhythmia; and no sustained ventricular arrhythmia was inducible in the remaining 45 (38%) patients. The latter two groups of 65 (55%) patients make up the study population. RESULTS: There were 16 deaths among the study population during a follow-up period of 25.3 +/- 19.6 months. The overall one- and two-year survival in these patients was 89% and 81%, respectively. No significant difference in survival was observed between patients with and without inducible VF (80% power to detect a fourfold survival difference). CONCLUSIONS: In 17% of patients with CAD and unexplained syncope, VF is the only inducible ventricular arrhythmia. Within the limits of this pilot study, long-term follow-up of patients with and without inducible VF demonstrates no difference in survival between the two groups. Therefore, the practice of ICD implantation in patients with CAD, unexplained syncope and inducible VF, especially with triple ventricular extrastimuli, may merit reconsideration. PMID- 11499728 TI - Characterization of the anatomy and conduction velocities of the human right atrial flutter circuit determined by noncontact mapping. AB - OBJECTIVES: This study was done to characterize human right atrial (RA) flutter (AFL) using noncontact mapping. BACKGROUND: Atrial flutter has been mapped using sequential techniques, but complex anatomy makes simultaneous global RA mapping difficult. METHODS: Noncontact mapping was used to map the RA of 13 patients with AFL (5 with previous attempts), 11 with counterclockwise and 2 with clockwise AFL. "Reconstructed" electrograms were validated against contact electrograms using cross-correlation. The Cartesian coordinates of points on a virtual endocardium were used to calculate the length and thus the conduction velocity (CV) of the AFL wave front within the tricuspid annulus-inferior vena cave isthmus (IS) and either side of the crista terminalis (CT). RESULTS: When clearly seen, the AFL wave front split (n = 3) or turned in the region of the coronary sinus os (n = 6). Activation progressed toward the tricuspid annulus (TA) from the surrounding RA in 10 patients, suggesting that the leading edge of the reentry wave front is not always at the TA. The IS length and CV was 47.73 +/- 24.40 mm (mean +/- SD) and 0.74 +/- 0.36 m/s. The CV was similar for the smooth and trabeculated RA (1.16 +/- 0.48 m/s and 1.22 +/- 0.65 m/s, respectively [p = 0.67]) and faster than the IS (p = 0.03 and p = 0.05 for smooth and trabeculated, respectively). CONCLUSIONS: Noncontact mapping of AFL has been validated and has demonstrated that IS CV is significantly slower than either side of the CT. PMID- 11499729 TI - Ablation of atrial tachycardia originating from the vicinity of the atrioventricular node: significance of mapping both sides of the interatrial septum. AB - OBJECTIVES: The purpose of the study was to examine the value of right- and left sided mapping to identify the site of tachycardia origin. BACKGROUND: Focal atrial tachycardia may originate from the vicinity of the atrioventricular node from either side of the interatrial septum. METHODS: In 16 patients undergoing radiofrequency catheter ablation of perinodal atrial tachycardia, activation mapping of the right and left side of the interatrial septum was performed. RESULTS: Atrial tachycardia originated from the right side of the interatrial septum in 10 patients (group A) and from the left side in 6 patients (group B). On the right side, earliest atrial activity preceded the onset of the P-wave by 49 +/- 15 ms in group A and by 38 +/- 8 ms in group B (NS), and it preceded the signal recorded from the right atrial appendage by 59 +/- 19 ms in group A and by 60 +/- 13 ms in group B (NS). On the left side, earliest activity preceded the onset of the P-wave by 27 +/- 16 ms in group A and by 51 +/- 6 ms in group B (<0.01), and it preceded the signal obtained from the right atrial appendage by 38 +/- 19 ms in group A and by 73 +/- 9 ms in group B (<0.01). Atrial tachycardias were successfully eliminated in all patients without impairment of atrioventricular conduction. During follow-up, two patients had a recurrence of tachycardia. CONCLUSIONS: Mapping of only the right side cannot exclude a left sided origin. Therefore, mapping of both sides of the interatrial septum is required prior to ablation of focal atrial tachycardia originating from the vicinity of the atrioventricular node. PMID- 11499730 TI - The clinical course of multifocal atrial tachycardia in infants and children. AB - OBJECTIVES: This study outlines the clinical course, treatment and the late outcome of infants and children with multifocal atrial tachycardia (MAT). BACKGROUND: Multifocal atrial tachycardia is defined by three distinct P waveforms, irregular P-P intervals, isoelectric baseline between P-waves and rapid rate on an electrocardiogram. Several smaller prior reports have described pediatric patients with MAT, but their long-term outcome has not been fully assessed. METHODS: The clinical records, echocardiograms and long-term follow-up of patients with MAT were reviewed and compared to previous reports of MAT. RESULTS: Fourteen boys and seven girls (median age 1.8 months) presented with MAT. At diagnosis, six patients had respiratory illness, of whom two were critical. Ten were asymptomatic. Seven patients had structural heart disease (SHD), one of whom died. Four of 15 patients (27%) with echocardiograms had diminished ventricular function. Ventricular rates were 111 to 253 beats/min (mean 181 beats/min). Median duration of the arrhythmia was 4.9 months (mean 6.7 months). Electrical cardioversion was attempted in 4 patients without success and 15 patients received antiarrhythmic medication. Seventeen patients were followed for a mean of 60 months. Four patients were lost to follow-up. There were no late arrhythmias. CONCLUSIONS: The majority of children with MAT are healthy infants under one year of age; a few may exhibit mild to life-threatening cardiorespiratory disease. Less often, MAT accompanies SHD. Mild ventricular dysfunction may be observed in the presence of MAT, but symptoms are few and resolution is generally complete. Response to antiarrhythmic agents is mixed, and cardioversion is of no avail. Finally, long-term cardiovascular and developmental outcome depends principally on underlying condition; for otherwise healthy children, it is excellent. PMID- 11499731 TI - Procedural outcomes and long-term survival among patients undergoing percutaneous coronary intervention of a chronic total occlusion in native coronary arteries: a 20-year experience. AB - OBJECTIVES: The study compared procedural outcomes and long-term survival for patients undergoing percutaneous coronary intervention (PCI) of a chronic total coronary artery occlusion (CTO) with a matched non-CTO cohort to determine whether successful PCI of a CTO is associated with improved survival. BACKGROUND: Percutaneous coronary intervention of a CTO is a common occurrence, and the long term survival for patients with successful PCI of a CTO has not been clearly defined. METHODS: Between June 1980 and December 1999, a total of 2,007 consecutive patients underwent PCI for a CTO. Utilizing propensity scoring methods, a matched non-CTO cohort of 2,007 patients was identified and compared to the CTO group. The cohorts were stratified into successful and failed procedures. RESULTS: The in-hospital major adverse cardiac event (MACE) rate was 3.8% in the CTO cohort. Technical success has improved over the last 10 years (overall 74.4%, slope 1.0%/yr, p = 0.02, R2 = 49.9%) as did procedural success (overall 69.9%, slope 1.2%/yr, p = 0.02, R2 = 51.5%) without a concomitant increase in in-hospital MACE rates (slope 0.1%/yr, p = 0.7). There was a distinct 10-year survival advantage for successful CTO treatment compared with failed CTO treatment (73.5% vs. 65.1%, p = 0.001). The CTO versus non-CTO 10-year survival was the same (71.2% vs. 71.4%, p = 0.9). Diabetics in the CTO cohort had a lower 10-year survival compared with nondiabetics (58.3% vs. 74.3%, p < 0.0001). CONCLUSIONS: These data represent follow-up of the largest reported series of patients undergoing PCI for a CTO. The 10-year survival rates for matched non-CTO and the CTO cohorts were similar. Success rates have continued to improve without an accompanying increase in MACE rates. A successfully revascularized CTO confers a significant 10-year survival advantage compared with failed revascularization. PMID- 11499732 TI - Geographical miss during catheter-based intracoronary beta-radiation: incidence and implications in the BRIE study. Beta-Radiation In Europe. AB - OBJECTIVES: We sought to determine the incidence and causes of geographical miss (GM) and to evaluate its impact on edge restenosis after intracoronary beta radiation therapy. BACKGROUND: Edge restenosis is a limitation of intracoronary beta-radiation therapy. Geographical miss is the situation in which the radiation source does not fully cover the injured segment and may lead to edge restenosis. METHODS: We analyzed 175 vessels treated according to the Beta-Radiation In Europe (BRIE) study protocol. The effective irradiated segment (EIRS) and both edges were studied with quantitative coronary angiography. The edges of the EIRS that were injured constituted the GM edges. Restenosis was defined as diameter stenosis >50% at follow-up. Geographical miss was determined by simultaneous electrocardiographic-matched, side-by-side projection of the source and balloons deflated at the injury site, in identical angiographic projections surrounded by contrast. RESULTS: Geographical miss affected 41.2% of the edges and increased edge restenosis significantly compared with non-GM edges (16.3% vs. 4.3%, respectively, p = 0.004). Restenosis was increased both in the proximal (p = 0.05) and distal (p = 0.02) GM edges compared with noninjured edges. Geographical miss associated with stent injury significantly increased edge restenosis (p = 0.006), whereas GM related to balloon injury did not significantly increase edge restenosis (p = 0.35). The restenosis in the EIRS was similar between vessels with and without GM (24.3% and 21.6%, respectively, p = 0.8). CONCLUSIONS: Geographical miss is strongly associated with restenosis at the edges of the EIRS. This effect is more prominent when caused by stenting. Geographical miss does not increase restenosis in the EIRS. PMID- 11499733 TI - Prognostic impact of diabetes mellitus in patients with heart failure according to the etiology of left ventricular systolic dysfunction. AB - OBJECTIVES: We sought to determine the relative impact of diabetes mellitus on prognosis in ischemic compared with nonischemic cardiomyopathy. BACKGROUND: Ischemic myocardium is characterized by increased reliance on aerobic and anaerobic glycolysis. Because glucose utilization by cardiomyocytes is an insulin mediated process, we hypothesized that diabetes would have a more adverse impact on mortality and progression of heart failure in ischemic compared with nonischemic cardiomyopathy. METHODS: We performed a retrospective analysis of the Studies Of Left Ventricular Dysfunction (SOLVD) Prevention and Treatment trials. RESULTS: In adjusted analyses, diabetes mellitus was strongly associated with an increased risk for all-cause mortality in patients with ischemic cardiomyopathy, (relative risk [RR] 1.37, 95% confidence interval [CI] 1.21 to 1.55; p < 0.0001), but not in those with nonischemic cardiomyopathy (RR 0.98, 95% CI 0.76 to 1.32; p = 0.98). The increased mortality in patients with ischemic cardiomyopathy compared with nonischemic cardiomyopathy was limited to those with ischemic cardiomyopathy and diabetes mellitus (RR 1.37, 95% CI 1.21 to 1.56; p < 0.0001). When patients with ischemic cardiomyopathy and diabetes mellitus were excluded, there was no significant difference in mortality risk between the ischemic and nonischemic cardiomyopathy groups after adjusted analysis (RR 0.99, 95% CI 0.86 to 1.15; p = 0.99). Previous surgical revascularization identified patients within the cohort with ischemic cardiomyopathy and diabetes mellitus, with improved prognosis. CONCLUSIONS: The differential impact of diabetes on mortality and heart failure progression according to the etiology of heart failure suggests that diabetes and ischemic heart disease interact to accelerate the progression of myocardial dysfunction. Evaluation of the potential for revascularization may be particularly important in patients with ischemic cardiomyopathy and diabetes mellitus. PMID- 11499734 TI - Cardiac nitric oxide production due to angiotensin-converting enzyme inhibition decreases beta-adrenergic myocardial contractility in patients with dilated cardiomyopathy. AB - OBJECTIVES: This study tested the hypothesis that angiotensin-converting enzyme (ACE) inhibitors attenuate beta-adrenergic contractility in patients with idiopathic dilated cardiomyopathy (DCM) through nitric oxide (NO) myocardial signaling. BACKGROUND: The ACE inhibitors increase bradykinin, an agonist of NO synthase (NOS). Nitric oxide inhibits beta-adrenergic myocardial contractility in patients with heart failure. METHODS: The study patients were given the angiotensin-1 (AT-1) receptor antagonist losartan for one week. The hemodynamic responses to intravenous dobutamine were determined before and during intracoronary infusion of enalaprilat (0.2 mg/min) with and without the NOS inhibitor N(G)-monomethyl-L-arginine (L-NMMA, 5 mg/min). RESULTS: In patients with DCM (n = 8), dobutamine increased the peak rate of rise of left ventricular pressure (+dP/dt) by 49 +/- 8% (p < 0.001) and ventricular elastance (Ecs) by 53 +/- 16% (p < 0.03). Co-infusion with enalaprilat decreased +dP/dt to 26 +/- 12% and Ecs to -2 +/- 17% above baseline (p < 0.05), and this anti-adrenergic effect was reversed by L-NMMA co-infusion (p < 0.05 vs. enalaprilat). In addition, intracoronary enalaprilat reduced left ventricular end-diastolic pressure (LVEDP), but not left ventricular end-diastolic volume, consistent with increased left ventricular distensibility. Infusion with L-NMMA before enalaprilat in patients with DCM (n = 5) prevented the reduction in +dP/dt, Ecs and LVEDP. In patients with normal left ventricular function (n = 5), enalaprilat did not inhibit contractility or reduce LVEDP during dobutamine infusion. CONCLUSIONS: Enalaprilat attenuates beta-adrenergic contractility and enhances left ventricular distensibility in patients with DCM, but not in subjects with normal left ventricular function. This response is NO modulated and occurs in the presence of angiotensin receptor blockade. These findings may have important clinical and pharmacologic implications for the use of ACE inhibitors, AT-1 receptor antagonists and their combination in the treatment of heart failure. PMID- 11499735 TI - Prognostic evaluation of neurohumoral plasma levels before and during beta blocker therapy in advanced left ventricular dysfunction. AB - OBJECTIVES: The study assessed the relative predictive potency of neurohumoral factors in patients with advanced left ventricular (LV) dysfunction during neurohumoral blocking therapy. BACKGROUND: The course of heart failure is characterized by progressive LV deterioration associated with an increase in cardiac (natriuretic peptides) and predominantly extracardiac (norepinephrine, big endothelin [big ET]) hormone plasma levels. METHODS: Plasma hormones were measured at baseline and months 3, 6, 12 and 24 in 91 patients with heart failure (left ventricular ejection fraction [LVEF] <25%) receiving 40 mg enalapril/day and double-blind atenolol (50 to 100 mg/day) or placebo. After the double-blind study phase, patients were followed up to four years. Stepwise multivariate regression analyses were performed with 10 variables (age, etiology, LVEF, symptom class, atenolol/placebo, norepinephrine, big ET, log aminoterminal atrial natriuretic peptide, log aminoterminal B-type natriuretic peptide [N-BNP] and log B-type natriuretic peptide [BNP]). During the study, the last values prior to patient death were used, and in survivors the last hormone level, New York Heart Association class and LVEF at month 24 were used. RESULTS: Thirty-one patients died from a cardiovascular cause during follow-up. At baseline, log BNP plasma level (x2 = 13.9, p = 0.0002), treatment allocation (x2 = 9.5, p = 0.002) and LVEF (x2 = 5.6, p = 0.017) were independently related to mortality. During the study, log BNP plasma level (x2 = 21.3, p = 0.0001) remained the strongest predictive marker, with LVEF (x2 = 11.2, p = 0.0008) log N-BNP plasma level (x2 = 8.9, p = 0.0027) and treatment allocation (x2 = 6.4, p = 0.0109) providing additional independent information. CONCLUSIONS: In patients with advanced LV dysfunction receiving high-dose angiotensin-converting enzyme inhibitors and beta blocker therapy BNP and N-BNP plasma levels are both independently related to mortality. This observation highlights the importance of these hormones and implies that they will likely emerge as a very useful blood test for detection of the progression of heart failure, even in the face of neurohumoral blocking therapy. PMID- 11499736 TI - Acquired growth hormone resistance in patients with chronic heart failure: implications for therapy with growth hormone. AB - OBJECTIVES: We aimed to determine whether growth hormone (GH) resistance is present in patients with chronic heart failure (CHF) and whether it may be linked to the biochemical response to GH treatment. BACKGROUND: Acquired GH resistance is a feature of severe illness, in particular, cachexia. In patients with CHF, the response to GH therapy appears to be variable. METHODS: Biochemical markers of the GH-insulin-like growth factor-I (IGF-I) axis were compared in 21 cachectic patients with CHF, 51 noncachectic patients and 26 healthy control subjects. In separate studies, the predictive value of baseline biochemical variables for the IGF-I response to GH treatment was analyzed. RESULTS: Cachectic patients showed an increase of total GH and immunologically intact GH (p < or = 0.0002) and a decrease of GH-binding protein (BP) (p = 0.005), IGF-BP3 (p = 0.01) and IGF-I (p = 0.06), compared with noncachectic patients. Similar changes were found when the cachectic group was compared with the control group. No differences were found between noncachectic patients and control subjects. Levels of GH-BP correlated with the IGF-I/GH ratio in all subgroups (all p < or = 0.002). Baseline GH-BP levels were related to the increase of IGF-I levels in response to GH treatment in patients with CHF after 24 h (r = 0.83, p = 0.005; n = 9; study 2), 44 days (r = 0.52, p = 0.007; n = 25; study 3) and 96 days (r = 0.54, p = 0.006; n = 24; study 3). CONCLUSIONS: Most cachectic and some noncachectic patients with CHF show features of acquired GH resistance. The principal predictors of the biochemical features of GH resistance and of the poor biochemical response to short-term and longer-term GH treatment are GH-BP plasma levels. The presence of GH resistance is potentially a major factor determining the response to GH therapy in patients with CHF. PMID- 11499737 TI - Predicting one-year mortality among elderly survivors of hospitalization for an acute myocardial infarction: results from the Cooperative Cardiovascular Project. AB - OBJECTIVES: We sought to develop a model based on information available from the medical record that would accurately stratify elderly patients who survive hospitalization with an acute myocardial infarction (AMI) according to their risk of one-year mortality. BACKGROUND: Prediction of the risk of mortality among older survivors of an AMI has many uses, yet few studies have determined the prognostic importance of demographic, clinical and functional data that are available on discharge in a population-based sample. METHODS: In a cohort of patients aged > or = 65 years who survived hospitalization for a confirmed AMI from 1994 to 1995 at acute care, nongovernmental hospitals in the U.S., we developed a parsimonious model to stratify patients by their risk of one-year mortality. RESULTS: The study sample of 103,164 patients, with a mean age of 76.8 years, had a one-year mortality of 22%. The factors with the strongest association with mortality were older age, urinary incontinence, assisted mobility, presence of heart failure or cardiomegaly any time before discharge, presence of peripheral vascular disease, body mass index <20 kg/m2, renal dysfunction (defined as creatinine >2.5 mg/dl or blood urea nitrogen >40 mg/dl) and left ventricular dysfunction (left ventricular ejection fraction <40%). On the basis of the coefficients in the model, patients were stratified into risk groups ranging from 7% to 49%. CONCLUSIONS: We demonstrate that a simple risk model can stratify older patients well by their risk of death one year after discharge for AMI. PMID- 11499738 TI - Risk prediction after myocardial infarction in the elderly. PMID- 11499739 TI - Time course and determinants of left ventricular function recovery after primary angioplasty in patients with acute myocardial infarction. AB - OBJECTIVES: We sought to evaluate the importance of time in relation to treatment, time course and determinants of recovery of left ventricular (LV) function in patients with acute myocardial infarction (AMI) undergoing primary percutaneous transluminal coronary angioplasty (PTCA). BACKGROUND: Myocardial salvage has been shown to be dependent on the time elapsed from the onset of AMI to reperfusion. METHODS: Left ventricular function was evaluated at hospital admission, after angioplasty, at 24 h and 6 months by both echocardiography and angiography and at 1, 7, 30, 90 and 180 days by echocardiography in 101 consecutive patients. RESULTS: Patients were allocated to three groups according to interval between symptom onset and angioplasty: <2 h (group A), 2 to 4 h (group B) and >4 h (group C). Patients in groups A and B showed a progressive improvement of LV function between day 7 and day 90, which became statistically significant at day 30 (p < 0.01). No LV function changes were noted in group C patients. Thrombolysis In Myocardial Infarction (TIMI) flow grade <3 at 24 h was not associated with any significant change in LV volume and function during the six-month follow-up period. Restenosis, when associated with TIMI flow grade 3 in the infarct-related vessel, did not influence LV function. Flow grade <3 of the infarct-related artery was not associated with any improvement of cardiac events independently from the time to treatment at the initial procedure. CONCLUSIONS: Patients undergoing primary PTCA for AMI have a good recovery of LV function if TIMI flow grade 3 is restored within 4 h. Coronary angioplasty limits further remodeling of the LV in patients treated after 4 h. PMID- 11499740 TI - Predictive factors for development of the no-reflow phenomenon in patients with reperfused anterior wall acute myocardial infarction. AB - OBJECTIVES: We sought to elucidate the clinical factors related to the development of no-reflow phenomenon after successful coronary reperfusion in patients with an acute myocardial infarction (AMI). BACKGROUND: Myocardial contrast echocardiography revealed that the no-reflow phenomenon is observed in some patients with a reperfused AMI, and those patients usually have poor functional and clinical outcomes. It is still unknown what clinical factors are related to the development of the no-reflow phenomenon. METHODS: Myocardial contrast echocardiography was performed 15 min after successful coronary reperfusion therapy in 199 patients with an anterior wall AMI who underwent successful coronary reperfusion with primary coronary angioplasty within 24 h after the onset of AMI. Multiple logistic regression analysis was used to identify independent predictors of the no-reflow phenomenon. RESULTS: Seventy nine patients showed the no-reflow phenomenon. Univariate analysis indicated that pre-infarction angina within 48 h before symptom onset, Killip class, Thrombolysis in Myocardial Infarction flow grade 0 on the initial coronary angiogram, the number of abnormal Q-waves and the wall motion score (WMS) on the echocardiogram obtained at hospital admission are related to the no-reflow phenomenon. Multivariate logistic regression analysis revealed that all of these factors, except for Killip class, are independent predictive factors of the no reflow phenomenon. CONCLUSIONS: Development of the no-reflow phenomenon is related to the severity of myocardial damage (number of Q-waves), the size of the risk area (WMS) and the occlusion status of infarct-related artery. In addition, ischemic preconditioning (pre-infarction angina) seems to be the factor that attenuates the no-reflow phenomenon. PMID- 11499741 TI - The prognostic value of troponin in patients with non-ST elevation acute coronary syndromes: a meta-analysis. AB - OBJECTIVES: This study was designed to compare the prognostic value of an abnormal troponin level derived from studies of patients with non-ST elevation acute coronary syndromes (ACS). BACKGROUND: Risk stratification for patients with suspected ACS is important for determining need for hospitalization and intensity of treatment. METHODS: We identified clinical trials and cohort studies of consecutive patients with suspected ACS without ST-elevation from 1966 through 1999. We excluded studies limited to patients with acute myocardial infarction and studies not reporting mortality or troponin results. RESULTS: Seven clinical trials and 19 cohort studies reported data for 5,360 patients with a troponin T test and 6,603 with a troponin I test. Patients with positive troponin (I or T) had significantly higher mortality than those with a negative test (5.2% vs. 1.6%, odds ratio [OR] 3.1). Cohort studies demonstrated a greater difference in mortality between patients with a positive versus negative troponin I (8.4% vs. 0.7%, OR 8.5) than clinical trials (4.8% if positive, 2.1% if negative, OR 2.6, p = 0.01). Prognostic value of a positive troponin T was also slightly greater for cohort studies (11.6% mortality if positive, 1.7% if negative, OR 5.1) than for clinical trials (3.8% if positive, 1.3% if negative, OR 3.0, p = 0.2) CONCLUSIONS: In patients with non-ST elevation ACS, the short-term odds of death are increased three- to eightfold for patients with an abnormal troponin test. Data from clinical trials suggest a lower prognostic value for troponin than do data from cohort studies. PMID- 11499742 TI - Noninvasive quantitative tissue characterization and two-dimensional color-coded map of human atherosclerotic lesions using ultrasound integrated backscatter: comparison between histology and integrated backscatter images. AB - OBJECTIVES: The purpose of the present study was to define clinicopathologically whether integrated backscatter (IB) combined with conventional two-dimensional echo (2DE) can differentiate the tissue characteristics of calcification (CL), fibrosis (FI), lipid pool (LP) with fibrous cap, intimal hyperplasia (IH) and thrombus (TH) and can construct two-dimensional tissue plaque structure in vivo. BACKGROUND: It is difficult to characterize the components of plaque using conventional 2DE techniques. METHODS: Integrated backscatter values of plaques were measured in the right common carotid and femoral arteries (total 24 segments) both during life and after autopsy in 12 patients (age 68 to 84 years, 10 men and two women). Integrated backscatter values were determined using a 5-12 MHz multifrequency transducer, setting the region of interests (ROIs) (11 x 11 pixels) on the echo tomography of the entire arterial wall (55 +/- 10 ROI/segment) and comparing it with histologic features in the autopsied arterial specimens. RESULTS: Corrected IB values obtained before death and at autopsy were significantly correlated (r = 0.93, p < 0.01). Corresponding to the histologic features, corrected IB values on the rectangle ROIs obtained during life were divided into five categories: category 1 (TH) 4 < IB < or = 6; category 2 (media and IH or LP in the intima) 7 < IB < or = 13; category 3 (FI) 13 < IB < or = 18, category 4 (mixed lesion) 18 < IB < or = 27 and category 5 (CL) 28 < IB < or = 33. In category 2, media and intima were differentiated using conventional 2DE. Under the above procedures, color-coded maps constructed with IB-2DE obtained during life precisely reflected the histologic features of media and intima. CONCLUSIONS: Integrated backscatter with 2DE represents a useful noninvasive tool for evaluating the tissue structure of human plaque. PMID- 11499743 TI - Tetrahydrobiopterin improves endothelial dysfunction in coronary microcirculation in patients without epicardial coronary artery disease. AB - OBJECTIVES: We aimed to determine whether intracoronary supplementation with nitric oxide (NO) synthase co-factor tetrahydrobiopterin (BH4) improves NO dependent coronary microvascular dilation in patients with coronary risk factors but no significant organic stenosis. BACKGROUND: Impaired coronary microvascular dilator reserve attributable to endothelial dysfunction plays an important role in the regulation of coronary blood flow (CBF). METHODS: Fifteen patients were measured for CBF (Doppler-wire and quantitative coronary angiography). Stimulated release of NO in the coronary microcirculation was evaluated by percent increase in CBF (%ACBF) at graded doses of intracoronary acetylcholine (1, 3, 10 and 30 microg/min). Measurements were repeated after intracoronary co-infusion of BH4 (4 mg/min) and acetylcholine. RESULTS: The patients were divided into two groups on the basis of CBF responses to acetylcholine: those with "diminished" (%deltaCBF <300%, n = 8) and "normal" (%deltaCBF >300%, n = 7) flow responses. Tetrahydrobiopterin significantly (p < 0.0001) improved acetylcholine-induced increases in CBF in patients with diminished flow responses, but exerted no effect in those with normal flow responses. Among the 15 studied patients, the magnitude of flow improvement by BH4 was inversely correlated with baseline flow responses (p < 0.02). Microvascular dilator response to direct NO donor (isosorbide dinitrate) was not affected by BH4. CONCLUSIONS: We demonstrated for the first time that intracoronary BH4 improved acetylcholine-induced microvascular dilator responses in patients with endothelial dysfunction in vivo. Thus, supplementation with BH4 may be a novel therapeutic means to increase NO availability for patients with coronary microvascular disease. PMID- 11499744 TI - Endothelium-dependent vasodilation is independent of the plasma L-arginine/ADMA ratio in men with stable angina: lack of effect of oral L-arginine on endothelial function, oxidative stress and exercise performance. AB - OBJECTIVES: This study was designed to determine the effect of two weeks' treatment with L-arginine on the ratio of plasma L-arginine to asymmetric dimethylarginine (ADMA), oxidative stress, endothelium-dependent vasodilatation to acetylcholine, exercise performance and heart rate variability in men with stable angina. BACKGROUND: The ratio of plasma L-arginine:ADMA has been proposed as a determinant of endothelium-dependent dilation; dietary supplementation with L-arginine has been shown to improve endothelium-dependent vasodilation and symptoms in some conditions. METHODS: Men (n = 40) with stable angina, at least one epicardial coronary artery with a stenosis >50% and a positive exercise test were randomized to receive L-arginine (15 g daily) or placebo for two weeks according to a double-blind parallel-group design. Plasma L-arginine, ADMA, 8-epi prostaglandin F2alpha (a marker of oxidative stress) and forearm vasodilator responses to brachial artery infusion of nitroprusside and acetylcholine (+/-L arginine) were measured. A standard Bruce protocol exercise test was performed before and at the end of the treatment period. RESULTS: Plasma L-arginine increased after oral L-arginine, whereas ADMA remained unchanged, leading to an increase in the L-arginine/ADMA ratio of 62 +/- 11% (mean +/- SE, p < 0.01). Despite a significant enhancement in acetylcholine response by intra-arterial L arginine at baseline, this response remained unchanged after oral L-arginine. Measures of oxidative stress and exercise performance after L-arginine/placebo were similar in placebo and active groups. CONCLUSIONS: In men with stable angina, an increase in plasma L-arginine/ADMA ratio after two weeks' oral supplementation with L-arginine is not associated with an improvement in endothelium-dependent vasodilatation, oxidative stress or exercise performance. PMID- 11499745 TI - Blood pressure reductions with exercise and sodium restriction in postmenopausal women with elevated systolic pressure: role of arterial stiffness. AB - OBJECTIVES: This study determined the relative efficacy of aerobic exercise (daily walking) and moderate dietary sodium restriction (sodium intake <100 mmol/day) for reducing systolic blood pressure (SBP) and pulse pressure (PP) in postmenopausal women with elevated initial levels, and the potential role of reductions in large artery stiffness in these changes. BACKGROUND: Lifestyle behaviors are recommended for lowering blood pressure (BP) in adults with elevated baseline levels, but there is little information as to the relative efficacy of different interventions or the mechanisms underlying their potential beneficial effects. METHODS: After baseline measurements and random assignment, 35 nonmedicated healthy postmenopausal women with SBP between 130 and 159 mm Hg completed three months of either aerobic (walking) exercise (n = 18; 62 +/- 9 years, mean +/- SD) or moderate dietary sodium restriction (SR) (n = 17; 65 +/- 10 years, mean +/- SD). RESULTS: Body mass and composition, plasma volume, and fasting concentrations of metabolic coronary risk factors did not differ between the groups at baseline or change with intervention. Systolic BP and PP at rest decreased with both exercise and SR (p < 0.05); however, the reductions were three- to fourfold greater with SR (p < 0.05). Sodium restriction, but not exercise, also reduced 24-h SBP and PP (p < 0.05). Aortic pulse wave velocity (PWV) and carotid augmentation index were reduced only with SR (p < 0.05). Changes in SBP and PP at rest and over 24 h correlated with the corresponding changes in aortic PWV (r = 0.53 to 0.61, p < 0.01). CONCLUSIONS: Moderate SR lowers SBP and PP in postmenopausal women with elevated baseline levels more than does daily walking. The greater blood pressure reductions with SR may be mediated in part by a decrease in the stiffness of the large elastic arteries. PMID- 11499746 TI - Relationship of the electrocardiographic strain pattern to left ventricular structure and function in hypertensive patients: the LIFE study. Losartan Intervention For End point. AB - OBJECTIVES: This study was designed to assess the relation of electrocardiographic (ECG) strain to increased left ventricular (LV) mass, independent of its relation to coronary heart disease (CHD). BACKGROUND: The classic ECG strain pattern, ST depression and T-wave inversion, is a marker for left ventricular hypertrophy (LVH) and adverse prognosis. However, the independence of the relation of strain to increased LV mass from its relation to CHD has not been extensively examined. METHODS: Electrocardiograms and echocardiograms were examined at study baseline in 886 hypertensive patients with ECG LVH by Cornell voltage-duration product and/or Sokolow-Lyon voltage enrolled in the Losartan Intervention For End point (LIFE) echocardiographic substudy. Strain was defined as a downsloping convex ST segment with inverted asymmetrical T-wave opposite to the QRS axis in leads V5 and/or V6. RESULTS: Strain occurred in 15% of patients, more commonly in patients with than without evident CHD (29%, 51/175 vs. 11%, 81/711, p < 0.001). When differences in gender, race, diabetes, systolic pressure, serum creatinine and high density lipoprotein cholesterol were controlled, strain on baseline ECG was associated with greater indexed LV mass in patients with (152 +/- 33 vs. 131 +/- 32 g/m2, p < 0.001) or without CHD (131 +/- 24 vs. 119 +/- 22 g/m2, p < 0.001). In logistic regression analyses, strain was associated with an increased risk of anatomic LVH in patients with CHD (relative risk 5.14, 95% confidence interval [CI] 1.16 to 22.85, p = 0.0315), without evident CHD (relative risk 2.91, 95% CI 1.50 to 5.65, p = 0.0016), and in the overall population when CHD was taken into account (relative risk 2.98, 95% CI 1.65 to 5.38, p = 0.0003). CONCLUSIONS: When clinical evidence of CHD is accounted for, ECG strain is likely to indicate the presence of anatomic LVH. Greater LV mass and higher prevalence of LVH in patients with strain offer insights into the known association of the strain pattern with adverse outcomes. PMID- 11499747 TI - Redilation of endovascular stents in congenital heart disease: factors implicated in the development of restenosis and neointimal proliferation. AB - OBJECTIVES: We sought to determine the incidence of and risk factors for the development of restenosis and neointimal proliferation after endovascular stent implantation for congenital heart disease (CHD). BACKGROUND: Risk factors for the development of restenosis and neointimal proliferation are poorly understood. METHODS: This was a retrospective review of patients who underwent endovascular stent redilation between September 1989 and February 2000. RESULTS: Of 368 patients who had 752 stents implanted, 220 were recatheterized. Of those 220 patients, 103 underwent stent redilation. Patients were classified into three groups: 1) those with pulmonary artery stenosis (n = 94), tetralogy of Fallot/pulmonary atresia (n = 72), congenital branch pulmonary stenosis (n = 9), status post-Fontan operation (n = 6), status post-arterial switch operation (n = 7); 2) those with iliofemoral venous obstruction (n = 6); and 3) those with miscellaneous disorders (n = 3). The patients' median age was 9.9 years (range 0.5 to 39.8); their mean follow-up duration was 3.8 years (range 0.1 to 10). Indications for stent redilation included somatic growth (n = 67), serial dilation (n = 27) and development of neointimal proliferation or restenosis, or both (n = 9). There was a low incidence of neointimal proliferation (1.8%) and restenosis (2%). There were no deaths. Complications included pulmonary edema (n = 1), hemoptysis (n = 1) and contralateral stent compression (n = 2). CONCLUSIONS: Redilation or further dilation of endovascular stents for CHD is effective as late as 10 years. The risk of neointimal proliferation (1.8%) and restenosis (2%) is low and possibly avoidable. Awareness of specific risk factors and modification of the stent implantation technique, including avoidance of minimal stent overlap and sharp angulation of the stent to the vessel wall and avoidance of overdilation, have helped to reduce the incidence of restenosis. PMID- 11499748 TI - Rapid evaluation of left ventricular volume and mass without breath-holding using real-time interactive cardiac magnetic resonance imaging system. AB - OBJECTIVES: The purpose of this study was to validate cardiac measurements derived from real-time cardiac magnetic resonance imaging (MRI) as compared with well-validated conventional cine MRI. BACKGROUND: Although cardiac MRI provides accurate assessment of left ventricular (LV) volume and mass, most techniques have been relatively slow and required electrocardiogram (ECG) gating over many heart beats. A newly developed real-time MRI system allows continuous real-time dynamic acquisition and display without cardiac gating or breath-holding. METHODS: Fourteen healthy volunteers and nine patients with heart failure underwent real-time and cine MRI in the standard short-axis orientation with a 1.5T MRI scanner. Nonbreath-holding cine MRI was performed with ECG gating and respiratory compensation. Left ventricular end-diastolic volume (LVEDV), left ventricular endsystolic volume (LVESV), ejection fraction (EF) and LV mass calculated from the images obtained by real-time MRI were compared to those obtained by cine MRI. RESULTS: The total study time including localization for real-time MRI was significantly shorter than cine MRI (8.6 +/- 2.3 vs. 24.7 +/- 3.5 min, p < 0.001). Both imaging techniques yielded good quality images allowing cardiac measurements. The measurements of LVEDV, LVESV, EF and LV mass obtained with real-time MRI showed close correlation with those obtained with cine MRI (LVEDV: r = 0.985, p < 0.001; LVESV: r = 0.994, p < 0.001; EF: r = 0.975, p < 0.001; LV mass: r = 0.977, p < 0.001). CONCLUSIONS: Real-time MRI provides accurate measurements of LV volume and mass in a time-efficient manner with respect to image acquisition. PMID- 11499749 TI - Regulation of myocardial betaARK1 expression in catecholamine-induced cardiac hypertrophy in transgenic mice overexpressing alpha1B-adrenergic receptors. AB - OBJECTIVES: Using a transgenic mouse model of myocardial-targeted overexpression of the wild-type alpha1B adrenergic receptor (AR) (Tg alpha43), we studied the role of the betaAR kinase (betaARK1) in the evolution of myocardial hypertrophy and its transition to heart failure (HF). BACKGROUND: Increased myocardial expression of betaARK1 has been shown to be associated with HF and certain models of hypertrophy. METHODS: Tg alpha43 mice and their nontransgenic littermate controls were treated with the alpha1AR agonist phenylephrine (PE) for 3, 7 or 14 days to characterize the cardiac consequences. RESULTS: Nontransgenic littermate control mice treated for 14 days with PE display cardiac hypertrophy with no increase in betaARK1 expression. However, Tg alpha43 animals show a reduced tolerance to 14-day PE treatment, demonstrated by reduced survival and severe cardiac hypertrophy. Moreover, PE treatment for three and seven days in Tg alpha43 mice resulted in an exaggerated hypertrophic response accompanied by significant cardiac biochemical abnormalities that are normally associated with HF, including fetal gene expression, reduced betaAR density and enhanced betaARK1 expression. We also found reduced myocardial stores of the sympathetic neurotransmitter neuropeptide Y. CONCLUSIONS: These data suggest that PE-treated Tg alpha43 mice have chronic activation of the cardiac sympathetic nervous system, which may be responsible for the appearance of apparent maladaptive hypertrophy with an evolution towards HF and sudden death. Thus, the cardiac phenotypes found in these mice are not the direct result of enhanced alpha1B AR signaling and suggest that betaARK1 is a key molecule in the transition of myocardial hypertrophy to HF. PMID- 11499750 TI - G protein-coupled receptor kinases: gotta real kure for heart failure? PMID- 11499751 TI - Nitric oxide synthase inhibitors decrease coronary sinus-free radical concentration and ameliorate myocardial stunning in an ischemia-reperfusion model. AB - OBJECTIVES: Our objective was to determine the effect of a nitric oxide synthase inhibitor, NG-nitro-L-arginine (L-NNA) on free radical generation and myocardial contractility after ischemia-reperfusion. BACKGROUND: Cardiotoxic free radicals are generated by ischemia-reperfusion sequences. Nitric oxide reacts with superoxide radical to form peroxynitrite, which generates additional free radicals. Our hypothesis was that by inhibiting NO production, free radical formation will be diminished, which should be cardioprotective. METHODS: We studied 32 dogs. Coronary occlusion-reperfusion (20 min each) sequences were created by intracoronary balloon angioplasty inflation-deflation. Using electron paramagnetic resonance, we monitored the coronary sinus concentration of ascorbate free radical (Asc*-), a measure of total oxidative flux. The L-NNA (4.8 mg/kg total) was infused intravenously during occlusion-reperfusion; control dogs received saline. Immunohistochemical staining demonstrated the peroxynitration product nitrotyrosine. RESULTS: In the control dogs Asc*- rose from 3.2 +/- SD 0.5 nmol/l to 4.8 +/- 1.1 nmol/l with reperfusion, a 50% rise. With L-NNA the Asc*- rose from 3.2 +/- 0.9 nmol/l to 4.0 +/- 1.2 nmol/l, a 25% rise (p < 0.01, L NNA vs. control). Echocardiographic left ventricular fractional area shortening (FAS) in the control dogs declined from 38 +/- 19% (baseline) to 26 +/- 14% (ischemia), and to 22 +/- 11% with reperfusion (p < 0.01 vs. baseline). With L NNA, FAS declined from 36 +/- 13% (baseline) to 27 +/- 12% (ischemia) but then rose to 33 +/- 14 with reperfusion (p = NS vs. baseline). Nitrotyrosine was present in the myocardium subjected to ischemia-reperfusion, but almost absent in dogs receiving L-NNA. Myocardial perfusion was not altered by L-NNA. CONCLUSIONS: The NO synthase inhibitors decrease coronary sinus free radical concentration and ameliorate myocardial stunning after ischemia-reperfusion. PMID- 11499752 TI - Validation of in vivo myocardial strain measurement by magnetic resonance tagging with sonomicrometry. AB - OBJECTIVES: This study was designed to validate strain measurements obtained using magnetic resonance tagging with spatial modulation of magnetization (SPAMM). We compared circumferential segment shortening measurements (%S) obtained using SPAMM to sonomicrometry %S in a canine model with (n = 28) and without (n = 3) coronary artery ligation. BACKGROUND: Magnetic resonance tagging enables noninvasive measurement of myocardial strain, but such strain measurements have not yet been validated in vivo. METHODS: Circumferential sonomicrometry crystal pairs were placed in apical myocardium at ischemic risk in ligation studies and in adjacent and remote myocardium. The %S was obtained from closely juxtaposed sonomicrometry and SPAMM sites. RESULTS: Paired data were available from 19 of 31 studies. Both methods distinguished remote from ischemic function effectively (p = 0.014 for SPAMM and p = 0.002 for sonomicrometry). SPAMM %S was similar to sonomicrometry %S in ischemic myocardium (2 +/- 3 vs. 0 +/- 3 p = 0.067) but was slightly higher than sonomicrometry %S in remote myocardium (11 +/- 10 vs. 7 +/- 5, p = 0.033). End-systolic (n = 30) and late systolic (n = 34) SPAMM %S correlated well with sonomicrometry %S (r = 0.84, p < 0.0001 and r = 0.88, p < 0.0001). CONCLUSIONS: Magnetic resonance tagging using SPAMM can quantitate myocardial strain in ischemic and remote myocardium. This study validates its application in scientific investigation and clinical assessment of patients with myocardial ischemia. PMID- 11499753 TI - Pressure rinsing of coronary stents immediately before implantation reduces inflammation and neointimal hyperplasia. AB - OBJECTIVES: This study evaluates whether rinsing stents with high pressure immediately before implantation minimizes stent-induced inflammation and neointimal formation. BACKGROUND: Several reports indicate that manual stent manipulation before implantation results in foreign body contamination and increased neointimal hyperplasia. METHODS: A stent-cleaning chamber was developed to rinse stents at a sustained hydrodynamic pressure of 4 atm for 10 s. Commercial pre-mounted stents were examined with different levels of manipulation: 1) untouched stents: no stent manipulation before implantation; 2) handled stents: manual stent re-crimping on the balloon; 3) rinsed stents: pressure-rinsed with the stent-cleaning chamber. In vitro surface analysis was evaluated by scanning electron microscopy. Neointimal hyperplasia and inflammation around stent struts were also assessed in the pig in-stent restenosis model. RESULTS: In vitro analysis revealed fewer contaminants on rinsed stents compared with untouched (p = 0.01) and handled stents (p < 0.001). In vivo, neointimal thickness, neointimal area and vessel percent stenosis were significantly reduced in rinsed, compared with not-rinsed, stents (p = 0.002, p = 0.007, p = 0.008 respectively). In addition, a significant reduction in the inflammatory infiltrate around struts was observed in untouched, compared with handled, stents (p = 0.04) and in rinsed, compared with not-rinsed, stents (p < 0.001). Regression analysis accounting for injury and neointimal thickness showed significant differences in slopes between "handled + not-rinsed" and "handled + rinsed" stents (p = 0.004), and between "untouched + not-rinsed" and "untouched + rinsed stents" (p = 0.037). CONCLUSIONS: Rinsing stents under high pressure immediately before coronary implantation results in less inflammation around struts and thinner neointima at 28 days in this pig model. PMID- 11499754 TI - Expression of exogenous tissue factor pathway inhibitor in vivo suppresses thrombus formation in injured rabbit carotid arteries. AB - OBJECTIVES: The aim of the present study was to test the hypothesis that retrovirus-mediated in vivo tissue factor pathway inhibitor (TFPI) gene transfer to the arterial wall would efficiently inhibit thrombosis without causing significant changes in systemic hemostatic variables. BACKGROUND: Acute coronary syndromes (unstable angina and acute myocardial infarction) are usually caused by atherosclerotic plaque rupture, with consequent activation of the coagulation cascade and circulating platelets. Tissue factor (TF) exposure represents an early event in this pathophysiologic sequence, leading to activation of the extrinsic coagulation pathway and thrombin formation. Tissue factor pathway inhibitor is a naturally occurring inhibitor of the extrinsic pathway. METHODS: In the present study, the gene coding for rabbit TFPI was inserted in a retroviral vector under control of a tetracycline-inducible promoter. Replication defective, infectious, recombinant retroviruses were used to transfect rabbit carotid arteries with either TFPI or a reporter gene--green fluorescent protein (GFP). RESULTS: Retroviral-mediated arterial gene transfer of TFPI resulted in potent inhibition of intravascular thrombus formation in stenotic and injured rabbit carotid arteries, whereas transfection of the contralateral carotid artery with GFP had no effect on thrombosis. No significant changes in systemic hemostatic variables (prothrombin time and partial thromboplastin time) were observed when thrombosis was inhibited. CONCLUSIONS: These data suggest that retroviral-mediated transfection of the arterial wall with TFPI might represent an attractive approach for the treatment of thrombotic disorders. PMID- 11499755 TI - Prevention of intimal hyperplasia with recombinant soluble P-selectin glycoprotein ligand-immunoglobulin in the porcine coronary artery balloon injury model. AB - OBJECTIVES: The role of P-selectin in the process of restenosis was evaluated using a recombinant immunoglobulin (Ig) chimera form of its ligand, soluble P selectin glycoprotein ligand-Ig (rPSGL-Ig), as a competitive inhibitor for the natural ligand on leukocytes. BACKGROUND: Inflammation and coagulation activation after vascular injury may be an important factor in the development of restenosis. P-selectin has been shown to mediate leukocyte-endothelium and leukocyte-platelet interaction. These interactions are mediated through binding of P-selectin to P-selectin glycoprotein ligand-1 (PSGL-1) located on the surface of leukocytes. METHODS: Balloon injury was induced in the left anterior descending and right coronary arteries of 16 pigs at a balloon/artery diameter ratio of 1.5:1. Either rPSGL-Ig (1 mg/kg) or saline was randomly administered 15 min before balloon injury as an intravenous bolus. Four weeks after injury, morphometric analysis, immunohistochemistry and histological evaluation were performed on injured arterial segments. RESULTS: Increased luminal area was found in the rPSGL-Ig group compared with the placebo group (1.63 +/- 0.57 mm2 vs. 1.26 +/- 0.32 mm2, p = 0.044) owing to significantly reduced neointimal hyperplasia (cross-sectional area, 0.46 +/- 0.45 mm2 vs. 0.13 +/- 0.11 mm2, p = 0.013). Immunohistochemistry and histological evaluation showed a significant decrease in the presence of tumor necrosis factor-alpha, interleukin-1 beta, and infiltration of macrophages in the injured vessel segments in the rPSGL-Ig group. CONCLUSIONS: P-selectin antagonism using rPSGL-Ig decreases neointimal hyperplasia following balloon injury, by inhibiting the inflammatory and thrombotic responses at the site of balloon injury, which appears to play a pivotal role in the pathogenesis of restenosis. PMID- 11499756 TI - HEART Group notification regarding "management of potential conflict of interest". Heart Editors Action Round Table. PMID- 11499757 TI - President's page: The changing face of research. PMID- 11499758 TI - Dental trauma in children presenting for treatment at the Department of Dentistry for Children and Orthodontics, Budapest, 1985-1999. AB - Data on children with dental trauma who presented for treatment at the Department of Dentistry for Children and Orthodontics in Budapest over a period of 15 years were analysed. The WHO guidelines were used to classify the traumatic injuries. A total of 590 children were involved, 810 teeth being affected. Children aged 7-14 years made up 88% of the cohort. The male:female ratio was 58:42. The permanent:primary ratio for the affected teeth was 90:10. The teeth most commonly affected were the maxillary central incisors. In 70% of the cases, only one tooth was traumatised. The incidence of dental trauma peaked at 10 years of age. The most common injury type observed was enamel-dentin crown fracture. The decreasing sequence of frequency of etiological factors was playing, sports, falls, cycling, road accidents and fighting. Of the accidents, 65% occurred at school or at home. Seventy seven per cent of the patients presented for medical care in the first 3 days after the accident. PMID- 11499759 TI - Prevalence of teeth with untreated dental trauma among nursery and primary school pupils in Dar es Salaam, Tanzania. AB - The aim of this study was to investigate the prevalence of teeth with untreated dental trauma among children aged 4-15 years in Dar es Salaam, Tanzania. A sample of 4524 children from three districts of different socio-economic status in the Dar es Salaam area was examined for signs of dental trauma in 1998/99. In 21% of all children examined, at least one type of untreated dental trauma was observed. A high percentage of untreated dental trauma was observed at the ages of 4 and 15 years. The most frequently observed type of dental trauma was enamel fracture (67%) followed by enamel-dentin fracture (26%). A significant gender influence on the occurrence of untreated dental trauma was observed, with more boys (23%) having untreated dental trauma than girls (19%). A higher percentage of untreated dental traumas was observed among children in the district with highest SES (26%) compared to the prevalence in the other two districts (14-17%). The findings of this study show that dental traumas are prevalent among Tanzanian children. Therefore, provision of information to the lay community about the importance of early management of dental trauma, ways of preventing trauma and appropriate handling of avulsed teeth is essential in order to minimize the rate of dental injuries and the related complications. PMID- 11499760 TI - Clinical application of computer-aided rapid prototyping for tooth transplantation. AB - The maintenance of healthy periodontal ligament cells in the donor tooth is one of the most important factors for successful tooth transplantation. This is achieved by minimizing the extraoral time during the surgical procedure. If a duplicate form of donor tooth could be obtained before extraction, it would be possible to precontour the recipient alveolar bone compatible with the donor tooth, and thereby reduce the extra-oral time of the donor tooth. We obtained a three-dimensional image with the real dimensions of the donor tooth from a CT Highspeed Advantage, allowing a life-sized resin model of the tooth to be fabricated. From 22 clinical cases, we achieved an average total transplantation time of 7.7 min. The average distance between the transplanted root surface and the alveolar bone from 12 available cases was 0.87 mm at the mesial cervix, 0.91 mm at the mesial apex, 0.98 mm at the distal cervix and 1.16 mm at the distal apex on the postoperative radiographs. Clinical data indicate that computer-aided rapid prototyping may be of value in minimizing the extra-oral time and possible injury to transplanted tooth during the process of autotransplantation. PMID- 11499761 TI - Effect of topical alendronate on root resorption of dried replanted dog teeth. AB - Alendronate (ALN) is a third generation bisphosphonate with demonstrated osteoclast inhibitory activity that may slow down the resorptive process after severe traumatic injuries. Eighty-two premolar roots of five mongrel dogs were endodontically treated and restored, extracted and treated as follows: 70 roots were bench dried for either 40 or 60 min. Thirty-eight of these roots were then soaked for 5 min in a 1 mM solution of ALN in Hanks' Balanced Salt Solution (HBSS) and replanted. Thirty-two roots were soaked for 5 min in HBSS and replanted. In the remaining 12 roots which were not exposed to the bench drying procedure, a 0.5 mM deep lingual mid-root cemental defect was made. Six of these roots were soaked in a 1 mM solution of ALN in HBSS for 5 min and replanted. The other six roots were soaked for 5 min in HBSS and replanted. Historical negative and positive controls were used from similarly treated teeth in our previous studies. After 4 months the dogs were killed and the roots prepared for histological evaluation. Five-microm-thick cross-sections of the root and surrounding tissue taken every 70 microm were evaluated for healing according to the criteria of Andreasen. In the 12 roots with cemental defects, healing with cementum of the damaged root surface was evaluated. In addition, residual root mass was also measured to determine the extent of root structure loss for each soaking method. Cemental healing took place in all 12 artificially damaged roots, indicating that these soaking media did not inhibit cementogenesis. The alendronate-soaked roots had statistically significantly more healing than the roots soaked in HBSS without alendronate. This improvement in healing was seen in all dogs except one and in all teeth except the first premolar. Soaking in alendronate also resulted in significantly less loss in root mass due to resorption compared to those teeth soaked in HBSS without alendronate. PMID- 11499763 TI - Treatment of replacement resorption with Emdogain--preliminary results after 10 months. AB - Ankylosis of traumatized teeth in children and adolescents may inhibit further development and growth of the corresponding jawbone. Therefore, ankylosed teeth should be removed. As an alternative treatment option to autotransplantation of a premolar, intentional replantation using Emdogain may be considered, provided the ankylosis is detected at an early stage or has only affected a small area of the root. Eleven ankylosed teeth presenting with replacement resorption were treated as follows: after tooth extraction, the root canal was treated extraorally and obturated by retrograde insertion of a titanium post. Emdogain was applied to the root surface and into the extraction socket with subsequent replantation of the tooth. During a mean follow-up period of 6.3 months, no signs of recurrence of ankylosis were noted. The horizontally and vertically measured Periotest scores were identical to those obtained on the adjacent teeth. These results suggest that intentional replantation using Emdogain may prevent or delay ankylosis of these replanted teeth. PMID- 11499762 TI - Periodontal healing after replantation of intentionally rotated teeth with healthy and denuded root surfaces. AB - The purpose of the study was to evaluate periodontal healing after replantation of intentionally rotated teeth with healthy and denuded root surfaces. Ten teeth with hopeless prognoses because of extensive alveolar bone loss and deep pockets extending to the apexes only on one or two surfaces, but with a healthy periodontal apparatus on the other surfaces, were selected. A mucogingival flap was elevated and the teeth were extracted carefully so as not to damage the healthy remnant of the periodontal membrane remaining on the root surfaces. Thorough extra-oral debridement was performed on the contaminated root surface and the remnant was carefully conserved. Retrofilling was also done to eliminate an endodontic cause of attachment loss. The teeth were horizontally rotated and then replanted so that the healthy root would face the connective tissue at the initially periodontally involved sites, and the root planed surfaces, which had been periodontally involved, would face the surfaces of the alveolar sockets at initially healthy sites. The teeth were splinted with adjacent teeth. Clinical parameters and radiographic examination were performed pre-operatively, and at 3 or 6 months, 1, 2, and 3 years postoperatively. One tooth was extracted 1.5 years postoperatively due to reduced support and the treatment strategy of the patient. The other nine teeth were well maintained without any discomfort for the whole maintenance period of 3 years. In all teeth, areas that previously had no pocketing but were now against denuded root surfaces maintained the previous pocket depth readings. In areas where a deep pocket was present but now had a root surface with healthy periodontium, a distinct decrease of mean pocket depth was observed at the first reexamination 3 months postoperatively. The mean radiographic alveolar bone level increased from 0.3% to 45.3% in 2 years and was thereafter maintained for the entire observation period. Loss of periodontal space and possible root resorption were observed in only one case without other ankylotic symptoms. These results suggest that the healing without ankylosis of an extensive denuded root surface may occur by mechanisms other than the maintenance of a viable periodontal ligament on the root surface. PMID- 11499764 TI - Extraction as a treatment alternative follows repeated trauma in a severely handicapped patient. AB - Handicapped patients with protruding maxillary incisors are prone to repeated dental trauma. A 13-year-old girl with cerebral palsy, severe mental retardation and seizure disorder was referred to our department for restoring the traumatized anterior teeth. Despite drug combination, the frequency of seizure attack was around 10 times a month. The oral examination showed multiple caries, gingival hyperplasia, class II malocclusion with 14 mm overjet and deep overbite. During the first 3 years of a 7-year follow-up period, six episodes of anterior tooth trauma due to seizure attack occurred. The trauma-related treatment performed included endodontic therapy, multiple composite restorations, apical repositional flap, and finally extraction of all four upper incisors with fabrication of a semi-fixed band-retained denture. The denture restored normal overbite and overjet with improved esthetics. For 4 years following the fabrication of denture, no trauma occurred to the anterior teeth in later seizure attacks. Considering inadequate control of seizure disorder, little ability of the patient to receive comprehensive orthodontic treatment, poor prognosis of restorations, and possible future injuries, the removal of non-functional, nonesthetic, trauma susceptible incisor teeth can be justified as an alternative to tooth preservation. PMID- 11499765 TI - Guidelines for the evaluation and management of traumatic dental injuries. PMID- 11499766 TI - High-energy illumination of the truth, or the serial endorser? PMID- 11499767 TI - Ask the experts. Enamel bonding. PMID- 11499768 TI - The science of communicating the art of esthetic dentistry. Part III: precise shade communication. AB - Shade matching is inherently complex and involves understanding the science of color, determining the shade and surface characteristics of teeth, and communicating this information to the laboratory technician. The laboratory technician then has the difficult task of mimicking the selected shade and ultimately reproducing it in the final restoration. PMID- 11499769 TI - Anterior esthetic rehabilitation on teeth and dental implants optimized with Procera technology: a case report. AB - With the latest developments of the Procera system, all-ceramic crowns have become an attractive solution to provide functional and esthetic rehabilitation on teeth and dental implants. The Procera AllCeram crown and Procera Abutment embrace the concept of computer-assisted design and computer-assisted machining (CAD/CAM) and can be used together for optimal esthetic result. The purpose of this case report was to illustrate the advantages of these new components for complex anterior rehabilitation. Three natural teeth and a Procera Abutment were restored using four Procera AllCeram crowns. Treatment planning and esthetic benefits are discussed. PMID- 11499770 TI - Periosteal connective tissue grafting or root coverage with enamel matrix derivative: a case report. AB - A 24-year-old female patient presented for the resolution of esthetic concerns associated with wide gingival recession facial to the maxillary left central incisor before orthodontic treatment. A periosteal connective tissue graft in conjunction with an enamel matrix derivative was used to achieve root coverage. This technique achieved healthy thick keratinized tissue coverage of the root surfaces and effectively improved the esthetics. These satisfactory clinical results have been maintained for 18 months, even through active orthodontic treatment. PMID- 11499771 TI - Comparison of color of resin composites of white and translucent shades with two shade guides. AB - PURPOSE: The purpose of this study was to compare the color of resin composites of white, translucent, and conventional shades with shade guides, based on the analysis of the color distribution of shade guides. The influence of color measuring mode, specular component included (SCI) or excluded (SCE), was also determined. MATERIALS AND METHODS: The labial portions of shade guide tabs (Vita and Chromascop) were polished flat with up to 2400-grit SiC paper. Color coordinates were measured according to CIE L*a*b* color scale on a reflection spectrophotometer with both the SCE and SCI modes. The color coordinates of white, translucent, and conventional shades of two brands of resin composites were measured and compared with those of the shade guides. RESULTS: There was no logical order in the color distribution of the two shade guides. The color of white and translucent shades of resin composites was located on the low CIE a* and CIE b* value area. The CIE L*, a*, b* values of resin composite with the same color designations were different, depending on the brand of material. The color difference (deltaE*) of shade guides by the measuring mode (SCE vs. SCI) was between 3.2 and 6.5. PMID- 11499772 TI - The clinical microscope in an aesthetic restorative practice. PMID- 11499773 TI - Visible light-curing: part II. PMID- 11499774 TI - The bottom line. PMID- 11499775 TI - Talking with patients. Ceramic inlays. PMID- 11499776 TI - Ultrasound of the knee. AB - Ultrasound is emerging as a viable imaging modality in the diagnosis and assessment of the musculoskeletal system. Advantages of ultrasound include its easy availability and multiplanar capability, as well as economic advantages. Unlike magnetic resonance imaging, ultrasound demonstrates the fibrillar microanatomy of tendons, ligaments and muscles, enhancing its diagnostic capability. The ability to compress. dynamically assess structures and compare easily with the contralateral side is advantageous. The patient's exact point of clinical tenderness can be correlated with underlying anatomical structures and associated pathology. The main strength of knee ultrasound is the assessment of para-articular disease. The specific structures best suited for ultrasound assessment include tendons, muscles and ligaments, as well as periarticular soft tissue masses. Joint effusions, synovial thickening, bursal fluid collections, intra-articular loose bodies, ganglion cysts, ligament and tendons tears, tendonitis and occult fractures can be diagnosed. With experience, ultrasound is a time-efficient, economical imaging tool for assessment of the knee. PMID- 11499777 TI - T1-weighted MR imaging for distinguishing large osteolysis of Paget's disease from sarcomatous degeneration. AB - OBJECTIVE: To report five symptomatic patients, four with unequivocal Paget's disease and large areas of osteolysis and one patient with presumed osteolytic Paget's disease, evaluated by MR imaging to confirm or exclude a sarcoma. DESIGN AND PATIENTS: Four men and one woman (median age 74 years) presented with new symptoms of pain. Four of these patients had unequivocal Paget's disease with large areas of osteolysis; one patient presented with large focal osteolysis and no other finding. MR imaging was performed in each case to exclude malignancy in the area of osteolysis. RESULTS: Two patients whose MR images showed a low signal abnormality on the T1-weighted sequence corresponding to osteolysis on the radiograph were found to have malignant degeneration. Three patients with osteolytic lesions on T1-weighted MR imaging showed preservation of fat signal in the areas of osteolysis, were not biopsied and have been free of malignant disease for from 12 months to 2 1/2 years. One patient had one area of osteolysis in the iliac bone which showed malignancy and another area of osteolysis which showed preservation of fat signal on the T1-weighted sequence. CONCLUSIONS: The information obtained from T1-weighted MR imaging sequences performed on patients with Paget's disease who have new symptoms and large areas of osteolysis could reliably be used in the clinical decision-making process between conservative follow-up and biopsy. PMID- 11499778 TI - Contrast-enhanced three-dimensional helical CT for soft tissue tumors in the extremities. AB - OBJECTIVE: To evaluate the ability of contrast-enhanced three-dimensional (3D) helical computed tomography (CT) to image soft tissue tumors in the extremities. DESIGN AND PATIENTS: Forty-five consecutive patients with soft tissue tumors in the extremities were examined (mean age 46.2 years; 24 females, 21 males). Twenty five patients had benign lesions and 20 had malignant lesions. All the patients underwent contrast-enhanced 3DCT scanning and magnetic resonance (MR) imaging preoperatively. All patients were surgically treated. Spiral CT scanning was performed with intravenous contrast enhancement. 3D reconstruction images were produced after thresholding, using Active-Windows (version 2.0, General Electric, Milwaukee, WI) software. 3DCT findings were compared in a masked fashion with the MR imaging and surgical findings regarding bone and major vessel invasion by the tumors. RESULTS: Forty-four of 45 tumors were satisfactorily imaged for the interpretation of their size, location and relationship to the skeleton and major vessels. One malignant tumor was judged on 3DCT to invade the major vessel, but the vessel proved to be normal at surgery. CONCLUSIONS: Contrast-enhanced 3D helical CT can be used for the evaluation of soft tissue tumors in the extremities, for preoperative surgical planning. PMID- 11499779 TI - Sonographic detection, evaluation and aspiration of infected acromioclavicular joints. AB - OBJECTIVE: To describe the sonographic findings of septic arthritis of the acromioclavicular joint. DESIGN AND PATIENTS: A retrospective study of five male patients was carried out. Four of the patients were referred because of signs and symptoms suggestive of glenohumeral joint septic arthritis, one for signs and symptoms suggestive of septic arthritis of the acromioclavicular joint. All the acromioclavicular joints were evaluated with ultrasound, aspirated and the aspirate cultured. RESULTS: All patients had normal ultrasound findings of their glenohumeral joints and distended acromioclavicular joints as determined by ultrasound. Ultrasound examination elicited focal tenderness over the acromioclavicular joint. Aspirates of each acromioclavicular joint grew pyogenic organisms. CONCLUSION: Infection in the acromioclavicular joint is uncommon, but is seen in increased frequency in immune-compromised patients and intravenous drug users. A normal glenohumeral joint on ultrasound in a patient suspected of having a septic shoulder should prompt careful review of the acromioclavicular joint. Aspiration of the acromioclavicular joint is easily performed under ultrasound guidance. PMID- 11499780 TI - Bony ankylosis following thermal and electrical injury. AB - OBJECTIVE: Bony ankylosis has been described following trauma, paralysis, psoriasis, Reiter's syndrome, ankylosing spondylitis, juvenile chronic arthritis and rheumatoid arthritis. Reports of bony ankylosis following thermal and electrical injury are limited. DESIGN AND PATIENTS: Thirteen cases of burn related joint ankylosis in four patients are presented. CONCLUSION: Patients with burns from thermal or electrical injury may develop bony ankylosis among other radiographic manifestations. This bony ankylosis may result either from bridging extra-articular heterotopic ossification with preservation of the underlying joint or from intra-articular fusion due to joint destruction. PMID- 11499781 TI - Quantitative assessment of an MR technique for reducing metal artifact: application to spin-echo imaging in a phantom. AB - OBJECTIVE: To quantify image artifact reduction using a new technique (MARS- metal artifact reduction sequence) in vitro. DESIGN: Coronal T1-weighted MR images were obtained through two metal phantoms (titanium/chromium-cobalt and stainless steel femoral prostheses) immersed in water. Comparison of artifact volume was made with images obtained using conventional and modified (MARS) T1 weighted sequences. Signal intensity values outside a range of +/-40% the average signal intensity for water were considered artifact and segmented into low or high signal artifact categories. Considering the arbitrary selection of this threshold value, volumetric calculations of artifact were also evaluated at +/ 50%, 60%, 70%, and 80% the mean signal for water. RESULTS: Conventional T1 weighted images produced 87% more low signal artifact and 212% more high signal artifact compared with the MARS modified T1-weighted images of the stainless steel prosthesis. Conventional T1-weighted images of the titanium prosthesis produced 84% more low signal artifact and 211% more high signal artifact than the MARS modified sequence. The level of artifact reduction was essentially uniform for the various threshold levels tested and was greatest at +/-20% the global signal intensity average for water. CONCLUSION: The MARS technique reduces the volume of image signal artifact produced by stainless steel and titanium/chromium cobalt femoral prostheses on T1-weighted spin-echo images in a tissue phantom model. PMID- 11499782 TI - Ossification of the ligamentum flavum associated with osteoblastoma: a report of three cases. AB - We report three cases of spinal osteoblastoma with ossification of the ligamentum flavum (OLF) adjacent to the tumor. The patients in this report, all young adults, had no symptoms except for back pain. Computed tomography (CT) demonstrated a typical radiolucent nidus in the spinal pedicle/lamina with a dense sclerotic rim. In addition, ectopic bone formation at the insertion point of the ligamentum flavum adjacent to the tumor was clearly illustrated. Magnetic resonance imaging (MRI) revealed the tumor and surrounding inflammatory responses, but OLF was not detected clearly. Histological examination revealed endochondral ossification of the ligamentum flavum that is quite unusual for normal young adults. Immunohistochemical assays in one case demonstrated that bone morphogenetic protein (BMP)-2/4 was expressed in the osteoblastic tumor cells. This case raises the possibility that BMPs secreted from the tumor cells triggered ectopic ossification in the spinal ligament. PMID- 11499783 TI - Pseudo-osteomyelitic crisis upon presentation of Gaucher disease. AB - We report on a 4-year-old boy adopted from Paraguay who presented with an acute onset of thigh pain. Initial clinical, imaging, and histopathologic findings suggested florid osteomyelitis. However, the development of pancytopenia on intravenous antibiotics prompted further investigation and the ultimate diagnosis of Gaucher disease. In retrospect, characteristic changes on conventional radiographic and MR images, as well as growth of a contaminant organism, pointed to the diagnosis of pseudo-osteomyelitis rather than osteomyelitis. PMID- 11499784 TI - Ultrasound-guided aspiration of posterior cruciate ligament ganglion cysts. AB - Intra-articular ganglion cysts of the knee joint are rare. Percutaneous aspiration of ganglion cysts of the knee, which has both diagnostic and therapeutic value, has been described, but usually under guidance by computed tomography (CT). We describe ultrasound-guided aspiration of posterior cruciate ligament cysts in two patients. PMID- 11499785 TI - Cotton-induced pseudotumor of the femur. AB - Twenty-five years prior to presentation a 41-year-old man had a femoral fracture stabilized with a 4.5 mm AO/ASIF steel plate. The femur healed uneventfully and the patient was asymptomatic for the following 20 years. He then noticed a slow growing swelling of the left thigh associated with a degree of weakness. Radiographs of the femur 25 years after fracture stabilization showed a massive expansive osteolytic process surrounded by a rim of bone. Magnetic resonance imaging (MRI) confirmed the presence of a large tumor. Since malignancy could not be excluded the patient underwent incisional biopsy. The histologic findings were nonspecific. Because of persistent symptoms the lesion was marginally excised. Intraoperatively a folded cotton sponge was found adjacent to the femur. Histopathologic investigation confirmed a foreign body reaction probably related to the retained cotton sponge. Reactive, foreign-body-induced change may mimic bone and or soft tissue malignancies. PMID- 11499786 TI - Focal fibrocartilaginous dysplasia: curettage as an alternative to conservative management or more radical surgery. AB - We describe two cases of focal fibrocartilaginous dysplasia, one treated conservatively while the other underwent curettage of the lesion. Resolution of tibia vara and healing of the focal fibrocartilaginous dysplasia was noted at 6 months in the patient who underwent curettage while the conservatively managed patient required 8 years of follow-up. Of the 17 cases of conservatively followed tibial focal fibrocartilaginous dysplasia described in the literature, 11 showed complete recovery after a median interval of 57 months. We believe that curettage may result in rapid healing by removing the persistent mesenchymal anlage which can interfere with the normal growth of the tibia. PMID- 11499787 TI - Absence of coupling between release and biosynthesis of peptide hormones in insect neuroendocrine cells. AB - Adipokinetic hormone (AKH)-producing cells in the corpus cardiacum of the insect Locusta migratoria represent a neuroendocrine system containing large quantities of stored secretory peptides. In the present study we address the question whether the release of AKHs from these cells induces a concomitant enhancement of their biosynthesis. The effects of hormone release in vivo (by flight activity) and in vitro (using crustacean cardioactive peptide, locustamyoinhibiting peptide, and activation of protein kinase A and C) on the biosynthetic activity for AKHs were measured. The intracellular levels of prepro-AKH mRNAs, the intracellular levels of pro-AKHs, and the rate of synthesis of (pro-)AKHs were used as parameters for biosynthetic activity. The effectiveness of in vitro treatment was assessed from the amounts of AKHs released. Neither flight activity as the natural stimulus for AKH release, nor in vitro treatment with the regulatory peptides or signal transduction activators appeared to affect the biosynthetic activity for AKHs. This points to an absence of coupling between release and biosynthesis of AKHs. The strategy of the AKH-producing cells to cope with variations in secretory stimulation seems to rely on a pool of secretory material that is readily releasable and continuously replenished by a process of steady biosynthesis. PMID- 11499788 TI - Yolk utilization in stick insects entails the release of vitellin polypeptides into the perivitelline fluid. AB - This study investigates the developmental fate of vitellin (Vt) polypeptides generated by limited proteolysis in an insect embryo. To this end, a number of polyclonal (pAb) and monoclonal antibodies (mAb) were raised against the yolk sac and the perivitelline fluid of late embryos of the stick insect Carausius morosus. Two dimensional immuno gel electrophoresis and Western blotting demonstrate that polypeptides resulting from Vt processing are present both in the yolk sac and the perivitelline fluid. At the confocal microscope, different labelling patterns were detected in the ooplasm depending on the stage of development attained by the embryo. At early developmental stages, label is associated with large unsegmented portions of the fluid ooplasm. During embryonic development, the fluid ooplasm is gradually transformed into yolk granules by intervention of vitellophages. Prior to dorsal closure, the yolk sac is separated from the perivitelline fluid by interposition of serosa cells (the so called serosa membrane). Several mAbs raised against the perivitelline fluid react specifically with this membrane suggesting that the release of Vt polypeptides from the yolk sac occurs by intracellular transit through the serosa cells. By immunocytochemistry, gold label appears associated with the cell surface and a number of vacuoles of the serosa membrane. These data are interpreted as suggesting that Vt polypeptides resulting from limited proteolysis in stick insect embryos are not exhaustively degraded within the yolk sac, but are instead transferred transcytotically to the perivitelline fluid through the serosa membrane. PMID- 11499789 TI - Azithromycin, a lysosomotropic antibiotic, impairs fluid-phase pinocytosis in cultured fibroblasts. AB - The dicationic macrolide antibiotic azithromycin inhibits the uptake of horseradish peroxidase (HRP) by fluid-phase pinocytosis in fibroblasts in a time- and concentration-dependent fashion without affecting its decay (regurgitation and/or degradation). The azithromycin effect is additive to that of nocodazole, known to impair endocytic uptake and transport of solutes along the endocytic pathway. Cytochemistry (light and electron microscopy) shows a major reduction by azithromycin in the number of HRP-labeled endocytic vesicles at 5 min (endosomes) and 2 h (lysosomes). Within 3 h of exposure, azithromycin also causes the appearance of large and light-lucentlelectron-lucent vacuoles, most of which can be labeled by lucifer yellow when this tracer is added to culture prior to azithromycin exposure. Three days of treatment with azithromycin result in the accumulation of very large vesicles filled with pleiomorphic content, consistent with phospholipidosis. These vesicles are accessible to fluorescein-labeled bovine serum albumin (FITC-BSA) and intensively stained with filipin, indicating a mixed storage with cholesterol. The impairment of HRP pinocytosis directly correlates with the amount of azithromycin accumulated by the cells, but not with the phospholipidosis induced by the drug. The proton ionophore monensin, which completely suppresses azithromycin accumulation, also prevents inhibition of HRP uptake. Erythromycylamine, another dicationic macrolide, also inhibits HRP pinocytosis in direct correlation with its cellular accumulation and is as potent as azithromycin at equimolar cellular concentrations. We suggest that dicationic macrolides inhibit fluid-phase pinocytosis by impairing the formation of pinocytic vacuoles and endosomes. PMID- 11499790 TI - Coordinated regulation of procollagens I and III and their post-translational enzymes by dissipation of mechanical tension in human dermal fibroblasts. AB - Mechanical tension governs fibroblast proliferation and survival and the homeostasis of the extracellular matrix to adapt its resistance to the mechanical requirements of the organs. To consolidate this view, we analysed the effect of tension release on the expression of molecules involved in the architecture and stabilisation of the collagen fibres, namely the procollagens type I and III, the amino- and carboxy-procollagen peptidases (N-pCP and C-pCP) and lysyl oxidase. Cells were cultured in conditions of high mechanical stress in monolayer on a collagen coat and under reduced tension by disruption of the cytoskeleton upon treatment with cytochalasin D in monolayer on a collagen coat or by integrin mediated stress relaxation in a freely retracting collagen gel. The mRNAs were measured by quantitative RT-PCR monitored by simultaneous reverse-transcription and amplification of an original internal standard. Tension relaxation resulted in a decreased expression of the procollagens type I and III, of the two expressed forms of C-pCP, of the two forms of N-pCP and of lysyl oxidase. Type III collagen, known to control diameter of the fibres, was less down-regulated than type I collagen. Interestingly, the expression of the two alternatively spliced forms of the N-pCP was dissimilarly regulated. These data suggest that mechanical tension may modulate the stiffness of the extracellular matrix by controlling not only the level of expression of its fibrillar constituents but also that of the enzymes participating in their extracellular processing and mechanical stabilisation. PMID- 11499791 TI - Targeted gene knockout of inner arm 1 in Tetrahymena thermophila. AB - Cilia and flagella contain at least eight different types of dynein arms. It is not entirely clear how the different types of arms are organized along the axoneme. In addition, the role each different type of dynein plays in ciliary or flagellar motility is not known. To initiate studies of dynein organization and function in cilia, we have introduced a mutation into one dynein heavy chain gene (DYH6) in Tetrahymena themophila by targeted gene knockout. We have generated mutant cells that lack wild-type copies of the DYH6 gene. We have shown that the DYH6 gene encodes one heavy chain (HC2) of Tetrahymena 18S dynein and that 18S dynein occupies the I1 position in the ciliary axoneme. We have also shown that Tetrahymena I1 is required for normal motility, normal feeding and normal doubling rate. PMID- 11499792 TI - Exclusion of pulmonary embolism using quantitative plasma D-dimer assays. AB - Recent quantitative studies using pulmonary angiography as reference method have indicated that the overall accuracy of quantitative D-dimer assays for the exclusion of pulmonary embolism (PE) in patients suspected of PE is poorer than was reported in earlier studies in which the same D-dimer assays were used (90 94% vs 98-100%). An explanation can be found in the fact that the earlier studies are hampered by the fact that the reference method was a compilation of clinical data and non-invasive diagnostic tests rather than a true gold standard. Furthermore, in those studies no discrimination was made between the milder cases of subsegmental PE and the more severe cases of segmental and larger PE. The lack of a true gold standard and preselection leading to reduced proportions of cases of subsegmental PE in the earlier studies rather than differences in the storage conditions of plasma samples or treatment with heparin, appear to have caused the discrepancies in the reported accuracy of D-dimer assays. It is concluded that the sensitivity and negative predictive values obtained with different quantitative D-dimer assays are in close agreement with each other, that is poor for subsegmental PE and excellent for segmental and larger PE. In diagnostic algorithms D-dimer can reliably exclude acute segmental and/or massive PE. Further work-up within 48 hours is still necessary in negative D-dimer outcomes to exclude subsegmental PE. PMID- 11499793 TI - Role of von Willebrand factor for the onset of arterial thrombosis. AB - Von Willebrand factor, a multimeric plasma protein released from either vascular endothelial cells or platelets, has been recently shown to play a crucial role in platelet thrombosis at sites exposed to high shear stress. Since VWF is only stored in two cellular components in normal adults, platelets and endothelial cells, the local release of VWF is an important regulatory mechanism of local thrombogenicities. Since endothelial cells are a larger source of plasma VWF, plasma VWF concentration can be used as an indicator of certain endothelial functions. Moreover, VWF is now being considered as a new target for the development of antithrombotic agents because the inhibition of VWF-mediated thrombosis may result in selective inhibition of platelet thrombosis at sites exposed to high shear stress, such as in atherosclerotic stenotic coronary arteries, without bleeding complications. PMID- 11499794 TI - Salivary HCV-antibodies; a follow-up cohort study of liver disease patients. AB - We have recently shown in Liver Clinic patients that saliva instead of serum may be used for anti-HCV detection. As compared to blood withdrawing, saliva is easier to obtain, non invasive, especially for infants. In the present study, sequential determination of serum and salivary anti-HCV was performed in the same cohort for 36 months. Anti-HCV seropositive and seronegative patients were studied. Blood and saliva samples were obtained simultaneously. From the anti-HCV seronegative patients (n=33), 161 sequential serum and 161 matched saliva samples were obtained. All were anti-HCV negative. From the anti-HCV seropositive patients (n=35), 131 sequential serum and 131 matched saliva samples were obtained. All sequential serum samples were anti-HCV positive. Of the saliva samples 126 (96%) were anti-HCV positive and five (4%) were anti-HCV negative. These five samples were obtained from two patients with autoimmune hepatitis and HCV-RNA seronegative by PCR. The results suggest that saliva may serve as a substitute for serum for the detection of anti-HCV antibodies. PMID- 11499795 TI - Estrogen and lipoproteins. PMID- 11499796 TI - Diagnostic evaluation of a new combined HIV p24 antigen and anti-HIV1/2/O screening assay. AB - To evaluate a new fourth generation assay for simultaneous detection of antibodies to the human immunodeficiency virus (HIV) 1 and 2 and HIV p24 antigen in daily routine we tested 675 sera obtained from 673 patients and compared the results to conventional antibody tests. In 546 uninfected patients the rate of unspecific reactivities was slightly higher in the new screening assay as compared to conventional antibody assays (1.1% vs. 0.4%). All 121 sera derived from patients with known HIV infection were detected correctly. In six patients from whom sera were obtained during early seroconversion the fourth generation ELISA was positive in three cases, while conventional third generation tests still were negative. In patients negative for HIV antibodies and low amounts of p24 antigen less than 100 pg/ml also the fourth generation ELISA remained negative. Thus, this new assay permits earlier detection of HIV infection and reduces the diagnostic window. It is a reliable tool for routine diagnosis of HIV, especially in blood donors and patients with high risk behavior. PMID- 11499797 TI - Validation of the first automated chemiluminescence protein-binding assay for the detection of 25-hydroxycalciferol. AB - BACKGROUND: Measurement of 25-hydroxyvitamin D, 25(OH)D is the ideal parameter to indicate the access of the organism to vitamin D. Numerous studies have shown that serum levels of 25(OH)D are the best markers of vitamin D deficiency, normal vitamin D supply or vitamin D intoxication. The aim of the study was to validate a beta-site version of the first fully automated chemiluminescence protein binding assay (CLPBA) for the detection of 25-hydroxycalciferol. METHODS: The newly developed CLPBA run on the Nichols Advantage Specialty Systems was compared to an inhouse radioimmunoassay (RIA), focussing on the major assay features such as imprecision, functional sensitivity, linearity, method comparison and suitability of serum or EDTA-plasma as well as establishing a preliminary reference range. RESULTS: Within-run imprecision is approximately 4.5% and total imprecision approximately 6% respectively (NCCLS protocol), functional sensitivity 6.8 microg/l. With mean recovery values of 96.9% and 98.7% for two diluted serum samples linearity is given over the measuring range. Due to different calibrations used for RIA and CLPBA the CLPBA reads approximately 70% lower (CLPBA = 0.321xRIA + 0.571, n = 469) but correlates well with the RIA (r = 0.9045). Method comparison with HPLC reveals a regression line of CLPBA = 0.8921xHPLC + 0.1358 (n = 54, r = 0.9117). Serum or EDTA-plasma is not equally suitable. Plasma samples read on average 5 microg/l higher than serum samples. The preliminary reference range is 11 microg/l to 84 microg/l (95% of all values). CONCLUSION: The validated 25(OH)D CLPBA is a very promising alternative to established commercially available 25(OH)D measurements and will, with its use on a fully automated platform, simplify the reliable quantification of 25 hydroxycalciferol significantly. PMID- 11499798 TI - The clinical importance of correctly diagnosing venous thrombosis. PMID- 11499799 TI - On difficulties in diagnosing deep vein thrombosis in daily clinical work and research practice. AB - This article enlightens the diagnostic difficulties when performing and interpreting a phlebography or color Doppler ultrasonography (CDU), which may have serious consequences in the daily clinical and laboratory work. In laboratory research as well as in clinical trials the most fundamental principle is to obtain a correct diagnosis. Less than 50% of patients with deep venous thrombosis (DVT) manifest the classical symptoms and signs of DVT and consequently the diagnosis is based upon methods such as phlebography or CDU. Some veins are especially hard to display, i.e. the veins of the planta pedis, the deep muscle veins of the calf and thigh and the deep internal iliac vein. Thus, the ideal diagnostic method must display the whole venous system from the planta pedis to the caval vein. A false negative examination puts the patient at risk of pulmonary embolism, which may present with a sudden onset and fatal outcome. A venous dysfunction as well as recurrent DVT will also be a continuous risk factor. A false positive interpretation may result in unnecessary anticoagulation treatment and bleeding complications. The combination of phlebography and CDU together with the strict use of a scoring method will increase the possibility of a proper diagnosis focusing on the diagnostically difficult areas of the venous system. PMID- 11499800 TI - Current issues in glycated haemoglobin measurement. PMID- 11499801 TI - Inhibitors of von Willebrand factor-cleaving protease in thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura (TTP), characterized by platelet thrombi in the arterioles and capillaries, is associated with antibodies that inhibit the activity of von Willebrand factor (vWF)-cleaving protease. Using a modified Bethesda method, we studied the inhibitor titers in patients who participated in the trial conducted by the Canadian Apheresis Group. Among the 41 patients investigated, the inhibitor titers at presentation were 1.4 +/- 1.7 U/mL (range 0.2-6.2 U/mL). Thirty-one patients (76%) had a titer > or = 0.2 U/mL, 8 patients (20%) had a titer > or = 2.0 U/mL but none had a titer > or = 10 U/mL. Among the 33 patients with an inhibitor titer < 2.0 U/mL (low titer group) and the 8 patients with an inhibitor titer > or = 2 U/mL (high titer group), 20 (61%) and 8 (100%) respectively had a platelet count < 25x10(9)/L (P = 0.04). Neurological abnormalities were among the presenting problems in 19 (58%) of the low titer and 6 (75%) of the high titer groups. Among the 23 patients who were randomized to plasma exchange, 5 patients had an inhibitor titer > or = 2 U/mL and none responded at the end of the first treatment cycle, while 8 of the 18 patients (44%) with a titer < 2 U/mL responded. This study shows that inhibitors of vWF cleaving protease are of low titers in most cases of acquired TTP. A higher inhibitor titer is associated with a more advanced stage of the disease and may delay the response to plasma exchange. PMID- 11499802 TI - Comparison of three methods for total homocysteine plasma determination. AB - Hyperhomocysteinemia is a well established risk factor for atherothrombotic disease, and the request for homocysteine determinations and the number of laboratories that need to perform this assay to assess individual risk profile is increasing. Different methods to evaluate homocysteine plasma levels are at present available. In the present study three methods, an in-house high-pressure liquid chromatographic (HPLC) method (considered as reference method) and two commercial immunoassays, an enzyme-linked immunoassay (EIA) and an automated fluorescence polarization immunoassay (FPIA), were used to measure homocysteine plasma levels in 100 samples. The median of homocysteine plasma levels obtained by HPLC was 9.0 micromol/L (range 4.2-23.0); the median of values obtained by EIA and FPIA were 10.6 micromol/L (range 3.3-21.5) and 9.6 micromol/L (4.8-20.2), respectively. The FPIA method showed the lowest within-run and between-run coefficients of variation (3.6% and 4.1%, respectively). There was a significant correlation between EIA and HPLC (r=0.81; p<0.0001), and between FPIA and HPLC (r=0.85; p<0.0001). The Bland-Altman analysis showed that FPIA agreed best with HPLC; EIA displayed a relatively wide scatter of difference data points. The present results indicate that the technological characteristics of the FPIA assay make this method suitable for the determination of Hcy in clinical laboratories. PMID- 11499803 TI - Stability of ketone bodies in serum in dependence on storage time and storage temperature. AB - The serum concentrations of beta-hydroxybutyrate and acetoacetate as well as the beta-hydroxybutyrate/acetoacetate ratio are important parameters for the differential diagnosis of certain inborn errors of metabolism. Acetoacetate, however, is an unstable compound which becomes rapidly decarboxylated. At a storage temperature of -20 degrees C about 40% of the acetoacetate is lost within 7 days and after 40 days storage at this temperature virtually all of the acetoacetate has become degraded. At -80 degrees C the decomposition of acetoacetate occurs with a much slower rate and only 15% of the initial acetoacetate is lost after 40 days storage. The rate constants for the decarboxylation reaction were found to be (6.4 +/- 2.9) * 10(-5) [min(-1)] at -20 degrees C and (0.4 +/- 0.3) * 10(-5) [min(-1)] at -80 degrees C. In contrast, beta-hydroxybutyrate is very stable during storage and hence should be used as main parameter for the evaluation of ketonemia. If determination of acetoacetate and/or of the beta-hydroxybutyrate/acetoacetate ratio is necessary, an assay immediately after collecting the serum samples is recommended. Otherwise, the serum samples should be frozen as soon as possible and stored at -80 degrees C during transport and storage. PMID- 11499804 TI - Analyzer transfer of a broad range high-sensitivity C-reactive protein immunoassay. AB - C-reactive protein (CRP) is measured in two main clinical situations: inflammation, where levels in the range of 5-300 mg/L are expected, and, more recently, assessment of the cardiovascular risk, where concentrations between 0.1 and 10 mg/L shoud be determined. Few commercially available methods display a measuring range covering both zones and laboratories are compelled to use two different assay protocols or even two different methods. The aim of the study was to adapt the Roche C-Reactive Protein (Latex) kit, initially developed for the Roche Cobas Integra analyzers, to the Hitachi Modular P800 analyzer in order to obtain on this instrument a broad range assay for CRP measurement. The method was successfully adapted and validated against a high-sensitivity and a traditional assay. The resulting method correlates well with the other two and displays a measuring range of 0.10-171 mg/L with an imprecision lower than 5.5%. This assay could be particularly practical in the routine clinical laboratory, being suitable for every use of CRP measurements. PMID- 11499805 TI - Foetal and neonatal thyroid disorders. PMID- 11499806 TI - Comparative analysis of IFN-gamma B7.1 and antisense TGF-beta gene transfer on the tumorigenicity of a poorly immunogenic metastatic mammary carcinoma. AB - Cancer progression is attributed in part to immune evasion strategies that include lack of co-stimulation, down-regulation of cell surface MHC molecules, and secretion of immunosuppressive factors, such as transforming growth factor beta (TGF-beta). Gene therapy has been employed to counter these mechanisms of immune evasion by transference of B7.1, IFN-gamma or antisense TGF-beta genes into tumor cells, resulting in cell surface expression of B7.1, upregulation of MHC class I and class II molecules, or elimination of tumor-derived TGF-beta, respectively. Although each of these transgenes has been shown to alter tumorigenicity in murine models, a direct comparison of their efficacy has not been performed. In this study, we have employed a very aggressive, poorly immunogenic and highly metastatic mammary model, 4T1, to compare the efficacy of B7.1, IFN-gamma and antisense TGF-beta gene transfer in stimulating an anti-tumor response. We demonstrate that both IFN-gamma and antisense TGF-beta gene expression significantly reduced the tumorigenicity of these cells compared to mock transduced cells, with IFN-gamma having a greater effect. In contrast, B7.1 gene transfer did not affect the tumorigenicity of 4T1 cells. The anti-tumor response directed against antisense TGF-beta-expressing 4T1 tumors was mediated by CD4+ and CD8+ T cells. However, CD8+ T cells, and not CD4+ T cells, appeared to mediate the anti-tumor response against IFN-gamma-expressing tumors. Treatment of tumor-bearing animals with IFN-gamma or antisense TGF-beta gene-modified tumor cell vaccines reduced the number of clonogenic metastases to the lungs and liver compared to treatment with mock-transduced cells. Finally, in a residual disease model in which the primary tumor was excised and mice were vaccinated with irradiated tumor cells, treatment of mice with vaccinations consisting of 4T1 cells expressing both antisense TGF-beta and IFN-gamma genes was the most effective in prolonging survival. PMID- 11499807 TI - Construction, expression and characterisation of a single-chain diabody derived from a humanised anti-Lewis Y cancer targeting antibody using a heat-inducible bacterial secretion vector. AB - A single-chain antibody fragment (scFv) of the humanised monoclonal antibody, hu3S193, that reacts specifically with Le(y) antigen expressed in numerous human epithelial carcinomas was constructed. A five-residue linker joined the C terminus of the V(H) and the N-terminus of the V(L), which prevented V-domain association into a monomeric scFv and instead directed non-covalent association of two scFvs into a dimer or diabody. The diabody was secreted into the E. coli periplasm using a heat-inducible vector, pPOW3, and recovered as a soluble, correctly processed protein, following osmotic shock or solubilised with 4 M urea from the insoluble fraction. The diabody from both fractions was isolated by a rapid batch affinity chromatography procedure, using the FLAG affinity tag to minimise degradation and aggregation. The purified diabody has an Mr of approximately 54 kDa, was stable and demonstrated similar binding activity as the parent monoclonal antibody, as measured by FACS and BIAcore analyses. The radiolabelled diabody showed a rapid tumour uptake, with fast blood clearance, proving it to be an excellent potential candidate as a tumour-imaging agent. PMID- 11499808 TI - Th1-cytokine induction and anti-tumor effect of 55 kDa protein isolated from Aeginetia indica L., a parasitic plant. AB - We have isolated a 55 kDa protein from the seed extract of Aeginetia indica L. (AIL), a parasitic plant, by affinity chromatography on an N-hydroxysuccinimide activated Sepharose High Performance column bound with F3, a monoclonal antibody that neutralizes the cytokine-inducing and anti-tumor effect of AIL. In the present study, we examined this protein (AILb-A) for cytokine induction and anti tumor effects by animal study, using syngeneic Meth-A tumor-bearing BALB/c mice, in which the Th2 response is genetically dominant. AILb-A administration resulted in markedly increased levels of Th1 cytokines [interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, IL-12 and IL-18] in the sera derived from Meth-A-bearing mice. The in vitro re-stimulation with AILb-A of splenocytes derived from AILb-A-primed mice also selectively induced Th1-type cytokines and antigen-specific killer cell activity. The neutralizing test using cytokine-specific antibodies revealed that AILb-A-induced IL-18 plays a most significant role for and killer cell-inducing activities. Furthermore, IL-12 and IL-18 induced by AILb-A inhibited specifically IL-10 and IL-4 production, respectively. Finally, we examined the anti-tumor effect of AILb-A in both Meth-A bearing BALB/c mice and Meth-A-bearing nude mice with BALB/c background. AILb-A exhibited a striking anti-tumor effect in normal BALB/c mice inoculated with Meth A cells. In athymic nude mice, the anti-tumor effect of AILb-A was relatively weak. These findings strongly suggested that AILb-A is a potent Th1 inducer and may be a useful immunotherapeutic agent for patients with malignant diseases. PMID- 11499809 TI - Renal carcinoma cell lines inhibit natural killer activity via the CD94 receptor molecule. AB - MHC class I molecules protect normal and transformed cells from lysis by natural killer (NK) cells through recognition of receptors expressed on leucocytes. Defects in NK cell activity and lymphokine activated killer (LAK) cell generation have been previously demonstrated in patients with renal cell carcinoma (RCC). However, to date, the importance of NK receptor/MHC class I interactions for immune evasion by RCC cells has not been described. In this study, human RCC cell lines (HTB46, HTB47, ACHN, CRL 1933 and HTB44) were found to be susceptible to lysis by both NK cells and interleukin-15 (IL-15)-derived LAK cells from normal donors in vitro. However, when NK cells were co-cultured with RCC cells their expression of the CD94 NK receptor molecule was significantly increased and their cytolytic activity against RCC targets was reduced. The cytolytic activity of NK cells was restored by the addition of IL-15, which further augmented the expression of CD94 on CD56+ NK cells. Disruption of NK receptor-MHC class I interactions by the addition of blocking antibodies to CD94 had no effect on the lysis of K562 or HTB47 targets by NK cells. However, the sensitivity of HTB46 cells to NK-mediated lysis was increased by blocking the CD94 receptor molecule, but only when the NK cells had not been previously co-cultured with RCC cells. This was independent of the presence of IL-15. These results show that RCC cells can inhibit NK activity via CD94 and suggest that disruption of interactions between receptor and ligand on RCC cells in vivo may augment the immune response against tumours by innate effector cells. PMID- 11499810 TI - Adoptive immunotherapy of cancer with pharmacologically activated lymph node lymphocytes: a pilot clinical trial. AB - Adoptive immunotherapy (AIT) of cancer with T lymphocytes may be limited by the need to activate tumor antigen-sensitized cells in vitro. In murine models, we have shown that AIT with tumor-sensitized T cells that have been pharmacologically activated with bryostatin 1 and ionomycin plus interleukin-2 can induce tumor regression. A Phase I clinical trial was carried out to assess the feasibility and toxicity associated with using tumor- or vaccine-draining lymph node cells, activated pharmacologically and expanded in culture with low dose interleukin-2 and infused intravenously, followed by IL-2 infusion. Nine patients were entered into the trial, and six were treated as planned. Average expansion of cell numbers over 13 to 27 days in culture was 118-fold. No patient's cells reached the target cell number (2.5 x 10(10)). Infusion of these cells did not result in any unexpected toxicities. The toxicities observed were related to IL-2 infusion, and conformed to the expected range of side-effects. Based on these Phase I results, additional trials, with tumor antigen vaccine sensitized DLN and technical modifications of the culture technique, are planned. PMID- 11499811 TI - Construction of humanized anti-ganglioside monoclonal antibodies with potent immune effector functions. AB - Gangliosides GD3, GD2 and GM2, which are the major gangliosides expressed on most human cancers of neuroectodermal and epithelial origin, have been focused on as effective targets for passive immunotherapy with monoclonal antibodies. We previously developed a chimeric anti-GD3 mAb, KM871, and a humanized anti-GM2 mAb, KM8969, which specifically bound to the respective antigen with high affinity and showed potent immune effector functions. Humanization of anti ganglioside antibody is expected to enhance its use for human cancer therapy. In the present study, we generated a chimeric anti-GD2 mAb, KM1138, and further developed the humanized form of anti-GD2 and anti-GD3 mAbs by the complementarity determining regions grafting method. The resultant humanized anti-GD2 mAb, KM8138, and anti-GD3 mAb, KM8871, showed binding affinity and specificity similar to those of their chimeric counterparts. In addition, both humanized mAbs had functional potency comparable to the chimeric mAbs in mediating the immune effector functions, consisting of antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity. The production of these humanized anti ganglioside mAbs, with potent effector functions and low immunogenicity, precedes the evaluation of the therapeutic value of anti-ganglioside mAbs in passive immunotherapy and the target validation for ganglioside-based vaccine therapy. PMID- 11499812 TI - Images and reflections from Mayo Clinic heritage. PMID- 11499813 TI - The growing impact of genetics on health care: do we have appropriate educational resources? PMID- 11499814 TI - Use of the Internet by patients and their families to obtain genetics-related information. AB - OBJECTIVE: To characterize use of the Internet by patients and their families referred to general genetics clinics. PATIENTS AND METHODS: We developed a survey to assess Internet use among patients visiting urban and rural clinics in Colorado and Wyoming. One hundred eighty-nine surveys were distributed to patients and their family members visiting outpatient general genetics clinics in spring 2000. The 8-page anonymous survey instrument asked about use of the Internet to obtain genetics-related information (GRI). All participants were asked whether a physician or health professional had referred them to the Internet for GRI. Subjects who had previously used the Internet to search for GRI were asked to rate whether they considered the GRI they encountered to be accurate, inaccurate, easy to understand, confusing, or trustworthy. RESULTS: One hundred fifty-seven surveys (83%) were returned (52% urban; 48% rural). Ninety (60%) of 149 respondents were at the clinic for a new-patient visit, and 59 (40%) were follow-up visits. All respondents were older than 17 years; 141 (91%) of 155 respondents were the patient's parent or guardian. Seventy-three (47%) of 155 respondents had searched the Internet for GRI prior to their clinic visit. The patients and families themselves initiated the majority of such efforts; only 8 (5%) of 148 respondents had been referred to a site on the World Wide Web by a physician. Interestingly, 136 (92%) of 147 respondents indicated that they would be likely to visit a Web site that was recommended by a geneticist. The most compelling reasons for searching the Internet for GRI were to get information in layperson's terms (60/131 [46%]); to get information about treatment (16/131 [12%]); and to get information about genetic research (16/131 [12%]). Among respondents who reported visiting GRI Web sites, 24 (41%) of 58 agreed that information was confusing or difficult to understand, 35 (53%) of 66 agreed that information was accurate and trustworthy, and 44 (77%) of 57 agreed that using the Internet was a positive experience. CONCLUSION: Internet use among patients referred to general genetics clinics and their family members appears to be widespread. Respondents reported that they found some of the information confusing and questioned its accuracy. Referral to Web sites by physicians was reported rarely, although the majority of respondents said they would visit a Web site recommended by a genetics physician. Further studies are needed to establish the accuracy of Internet information and how best to integrate and/or accommodate the data into the genetics clinic. PMID- 11499815 TI - Prevalence and patterns of presentation of genetic disorders in a pediatric emergency department. AB - OBJECTIVES: To determine the prevalence and patterns of presentation of previously diagnosed and of suspected genetic disorders among pediatric emergency department (ED) visits to a hospital that serves an inner-city population. PATIENTS AND METHODS: A retrospective review of 15,258 pediatric (<18 years old) ED visits at Lincoln Medical and Mental Health Center was undertaken for visits that occurred between October 1998 and February 1999. Suspected genetic disorders, classified into chromosomal, single gene, multifactorial, and other syndromic categories, were recorded. RESULTS: Of 15,258 visits reviewed, 2839 visits (18.6%) were by patients who had known or suspected genetic disorders. Previously diagnosed genetic disorders were documented in 80 visits (2.8%). Of these, 69 visits (86.2%) were related to single gene disorders, 3 (3.8%) to chromosomal disorders, 6 (7.5%) to multifactorial disorders, and 2 (2.5%) to disorders in the "other" category. Of these 80 visits, 59 (74%) were associated with sickle cell disease. The remaining 2759 visits (97.2%) were associated with complaints or diagnoses that suggested the possibility of an underlying genetic disorder requiring further evaluation and diagnostic work-up. CONCLUSIONS: Pediatric patients with known or suspected genetic disorders are frequently treated in EDs. Awareness of underlying genetic disorders facilitates diagnostic evaluation, treatment planning, and referral to a genetics clinic for counseling. PMID- 11499816 TI - Effect of abciximab on the outcome of emergency coronary artery bypass grafting after failed percutaneous coronary intervention. AB - OBJECTIVE: To evaluate the outcome of coronary artery bypass grafting (CABG) for failed percutaneous coronary intervention (PCI) in patients who had received abciximab. PATIENTS AND METHODS: In this retrospective study, we analyzed the records of patients who had PCI at our institution between January 1994 and December 1998 and identified those who had urgent or emergency CABG within 48 hours after PCI. CABG was performed for failed PCI in patients who had ongoing ischemia, hemodynamic compromise, or both. These patients were categorized into 2 groups depending on whether they had been given abciximab during PCI. We compared blood product transfusion requirements, bleeding complications, and frequency of in-hospital adverse events of the 2 groups. RESULTS: Of 5636 patients who had PCI, 77 (1.4%) had urgent or emergency CABG within 48 hours, including 11 who were given abciximab (abciximab group) during PCI and 66 who were not given abciximab (no abciximab group). The 2 groups had similar baseline characteristics. The mean +/- SD time to surgery was 8.4 +/-8.0 hours (median, 6 hours) for the abciximab group vs 12.1 +/- 12.5 hours (median, 4 hours) for the no abciximab group. Major bleeding (Thrombolysis in Myocardial Infarction criteria) occurred in 9 (90%) of 10 patients in the abciximab group vs 48 (77%) of 62 patients in the no abciximab group. The total volumes of intraoperative autotransfusion and transfusion of red blood cells and fresh frozen plasma tended to be higher for the abciximab group. Also, this group received a mean of 13.9 U of platelets vs 3.2 U for the no abciximab group (P<.001). However, no in hospital deaths occurred among patients in the abciximab group, and adverse events were infrequent and comparable between the 2 groups. No difference was noted between the 2 groups in the frequency of surgical reexploration for bleeding. CONCLUSION: Transfusion requirements are higher for patients who undergo emergency or urgent CABG after having received abciximab during PCI. However, in-hospital adverse events are infrequent and comparable to those for patients who do not receive abciximab. PMID- 11499817 TI - Hypopituitarism caused by intrasellar aneurysms. AB - OBJECTIVE: To determine the prevalence, clinical presentation, and outcome of hypopituitarism due to an intrasellar aneurysm. PATIENTS AND METHODS: We performed a retrospective review of Mayo Clinic, Rochester, Minn, medical records from 1950 through 1995. We calculated the prevalence of hypopituitarism, characterized the clinical presentation, and evaluated postoperative outcomes. RESULTS: Of 4087 patients with a diagnosis of hypopituitarism, 7 had hypopituitarism due to an intrasellar aneurysm, accounting for a prevalence of 0.17%. Adrenal, thyroid, and gonadal deficiencies were observed in 7, 6, and 5 patients, respectively. The prolactin level was increased in the 2 patients in whom it was measured. No patient had diabetes insipidus. All had visual impairment. In 5 patients an intrasellar aneurysm of the internal carotid artery was diagnosed preoperatively based on 1 or more imaging procedures. In the other 2 patients (1 with an internal carotid artery aneurysm and 1 with an anterior cerebral artery aneurysm), the diagnosis was made at surgical exploration. Four patients underwent a surgical procedure: 2 had direct packing of the aneurysm, and 2 had a bypass with proximal carotid occlusion or aneurysm trapping. Pituitary function remained unchanged postoperatively. CONCLUSION: An intrasellar aneurysm is a rare cause of hypopituitarism (0.17% of cases). Preoperative diagnosis is essential for successful surgical outcomes. Hypopituitarism is usually permanent. PMID- 11499818 TI - Utility of endoscopic ultrasonography in endoscopic drainage of pancreatic pseudocysts in selected patients. AB - OBJECTIVES: To determine the effect of endoscopic ultrasonography (EUS) on endoscopic drainage of pancreatic pseudocysts and to determine patency with fistula dilation and placement of multiple stents. PATIENTS AND METHODS: Between September 1995 and January 1999, 19 patients underwent endoscopic drainage of pancreatic pseudocysts, 17 of whom were assessed by EUS before drainage. Radial EUS scanning was used to detect an optimal site of apposition of pseudocyst and gut wall, free of intervening vessels. A fistula was created with a fistulatome, followed by balloon dilation of the fistula tract. Patency was maintained with multiple double pigtail stents. The primary goal of this retrospective study was to determine whether EUS affected the practice of endoscopic drainage of pancreatic pseudocysts. RESULTS: In 3 patients, drainage was not attempted based on EUS findings. In the other 13 patients (14 pseudocysts), creation of a fistula was successful on 13 occasions, and no immediate complications occurred. However, 1 patient subsequently developed sepsis that required surgery. All other patients were treated with balloon dilation, multiple stents, and antibiotics, with no septic complications. Of 14 pseudocysts (in 13 patients), 13 (93%) resolved. CONCLUSIONS: Results of EUS may alter management of patients considered for endoscopic drainage of pancreatic pseudocysts. Endoscopic ultrasonography was useful for selecting an optimal and safe drainage site. The combination of balloon dilation, multiple stents, and antibiotics appears to resolve pancreatic pseudocysts without septic complications. PMID- 11499819 TI - Anatomy of the first septal perforating artery: a study with implications for ablation therapy for hypertrophic cardiomyopathy. AB - OBJECTIVE: To determine the variability in the size and distribution of the first septal perforating artery (FSPA). MATERIAL AND METHODS: In this pilot study, 10 fresh autopsy hearts from patients who did not have hypertrophic cardiomyopathy (HCM) or clinical evidence of coronary artery disease were evaluated for the variability in the size of the FSPA. The size of the FSPA was also measured during coronary angiography in 8 patients with HCM who were undergoing alcohol septal ablation. RESULTS: Of the 10 autopsy hearts, 2 had a large FSPA (> or = 1.0 mm in maximal diameter) with prominent septal myocardial distribution, 2 had a medium-sized FSPA (0.5-0.9 mm), 2 had a small FSPA (0.1-0.4 mm), 3 had a tiny FSPA (< 0.1 mm), and 1 had an indiscernible ostium. In 2 patients the FSPA supplied the right ventricular free wall. In 4 patients the basal ventricular septum was incompletely supplied by the FSPA. Of the 8 patients with HCM, the FSPA was larger than 2 mm in diameter in 2 patients, 1 to 2 mm in 4, and smaller than 1 mm in 2. The distance between the left anterior descending coronary artery ostium and the origin of the FSPA ranged between 13.1 and 37.4 mm, indicating a large variation in the size and distribution of the FSPA. CONCLUSIONS: Variability in the size and distribution of the FSPA in patients without HCM was substantial. Areas of the heart other than the basal septum were supplied in some patients by the FSPA. In other patients the FSPA did not supply the entire basal septum. Similar findings were noted in patients with HCM. A detailed evaluation of the distribution of the FSPA may be necessary in all patients with HCM who are undergoing alcohol septal ablation. PMID- 11499820 TI - Left ventricular and biventricular pacing in congestive heart failure. AB - Dual-chamber pacing improved hemodynamics acutely in a subset of patients with left ventricular (LV) dysfunction but conveyed no long-term symptomatic benefit in most. More recently, LV pacing and biventricular (multisite) pacing have been used to improve systolic contractility by altering the electrical and mechanical ventricular activation sequence in patients with severe congestive heart failure (CHF) and intraventricular conduction delay or left bundle branch block (LBBB). Intraventricular conduction delay and LBBB cause dyssynchronous right ventricular and LV contraction and worsen LV dysfunction in cardiomyopathies. Both LV and biventricular cardiac pacing are thought to improve cardiac function in this situation by effecting a more coordinated and efficient ventricular contraction. Short-term hemodynamic studies have shown improvement in LV systolic function, which seems more pronounced with monoventricular LV pacing than with biventricular pacing. Recent clinical studies in limited numbers of patients suggest long-term clinical benefit of biventricular pacing in patients with severe CHF symptoms. Continuing and future studies will demonstrate whether and in which patients LV and biventricular pacing are permanently effective and equivalent and which pacing site within the LV produces the most beneficial hemodynamic results. PMID- 11499821 TI - Pathophysiology, diagnosis, and current management strategies for chest pain in patients with normal findings on angiography. AB - Chest pain syndromes in patients with normal angiographic findings represent a multifactorial pathophysiologic state, which may range from abnormalities in pain perception to abnormalities in endothelial- and non-endothelial-dependent coronary flow reserve associated with myocardial ischemia. Treatment begins with an accurate diagnosis by obtaining a comprehensive history and performing a physical examination, followed possibly by performing functional angiography in those who continue to have symptoms. This approach may help to determine appropriate treatment. PMID- 11499822 TI - Making sense of refractive surgery in 2001: why, when, for whom, and by whom? AB - Surgical alteration of the focusing or refractive properties of the eye has been performed on millions of patients. An array of procedures to correct myopia, hyperopia, astigmatism, and presbyopia have been introduced over the past 25 years with varying degrees of success. Improved technology has increased patient and physician satisfaction and enthusiasm. Currently available surgical procedures can be categorized as incisional, surface-altering, lamellar, and intraocular. The choice of procedure depends on individual patient indications and contraindications based on results of ocular examinations, eg, corneal pachymetry to measure corneal thickness, keratometry to measure the corneal curvature, basal tear secretory rate, and dark-adapted pupil size. The postoperative uncorrected visual acuity depends, in large part, on the quality of the preoperative evaluation and refraction. Before scheduling a patient for surgery, the ophthalmologist must ensure that the patient understands the potential risks of the procedure and has realistic expectations for the postoperative level and quality of uncorrected visual acuity. Postoperative complications include corneal flap displacement, undercorrection and overcorrection, and epithelial ingrowth under the corneal flap and inflammatory keratitis. Postoperative dry eye, infection, and inflammation are usually treated medically. Ongoing technological innovations to customize the surgical approach to an individual patient's eye continue to improve outcomes. PMID- 11499823 TI - Karl Landsteiner--discoverer of the major human blood groups. PMID- 11499824 TI - Medical advice for international travelers. AB - Each year, approximately 30 to 40 million Americans travel outside the United States. Although the most popular destinations are Europe, Central America, and the Caribbean, travel to Africa and Asia is increasing substantially. International travel, particularly to developing countries, can be associated with the risk of infectious and noninfectious diseases. These risks can be decreased, eliminated, or modified with vaccinations, prophylactic medications, and education. Optimally, pretravel advice must be individualized to a person's medical history, itinerary, and risk behavior. In addition to risk assessment based immunizations, issues such as traveler's diarrhea, malaria prophylaxis, sexually transmitted diseases, and management of underlying medical problems must form a part of pretravel management. Adventure or prolonged travel or persons with underlying medical diseases such as insulin-dependent diabetes mellitus, transplantation, immunodeficiencies, and dialysis warrant additional preventive measures. This review primarily updates pretravel management of adults. PMID- 11499825 TI - 29-year-old man with fever, malaise, and abdominal pain. PMID- 11499826 TI - Cutaneous lymphoma associated with Epstein-Barr virus infection in 2 patients treated with methotrexate. AB - Whether patients with rheumatoid arthritis (RA) have an increased risk of developing non-Hodgkin lymphoma is controversial, and opinions differ on the possible role of methotrexate in the occurrence of lymphomas in patients with RA. We report 1 T-cell lymphoma and 1 B-cell lymphoma restricted to the skin associated with Epstein-Barr virus infection that healed completely and spontaneously after discontinuation of methotrexate in a man with RA and a woman with dermatomyositis. Cutaneous infiltrating cells were infected by a replicative form of Epstein-Barr virus. After discontinuation of methotrexate, the cutaneous lesions disappeared completely in 15 days without recurrence. Discontinuation of methotrexate is necessary in patients with RA or dermatomyositis who have a lymphoproliferative disorder, and a follow-up period of several weeks should be observed before specific therapy is initiated. PMID- 11499827 TI - Arteritis and brachial plexus neuropathy as delayed complications of radiation therapy. AB - Radiation-induced arteritis of large vessels and brachial plexus neuropathy are uncommon delayed complications of local radiation therapy. We describe a 66-year old woman with right arm discomfort, weakness, and acrocyanosis that developed 21 years after local radiation for breast adenocarcinoma. Arteriography revealed arteritis, with ulcerated plaque formation at the subclavian-axillary artery junction, consistent with radiation-induced disease, and diffuse irregularity of the axillary artery. Electromyography showed a chronic brachial plexopathy. The patient's acrocyanosis, thought to be due to digital embolization from her vascular disease, improved with antiplatelet therapy. The concurrent combination of radiation-induced arteritis and brachial plexopathy is uncommon but should be considered in patients presenting with upper extremity pain or weakness after radiation therapy. PMID- 11499828 TI - External-beam radiation therapy in the treatment of diffuse tracheobronchial amyloidosis. AB - Tracheobronchial amyloidosis is characterized by deposits of amyloid in airway walls. No effective treatment is known. We describe a 59-year-old woman who presented with increasing symptoms of airway obstruction due to diffuse deposition of amyloid throughout her tracheobronchial tree. She was treated with external-beam radiation therapy (20 Gy) with marked improvement in her symptoms, effort tolerance, bronchoscopic appearance, and forced expiratory volume in 1 second (1.39 L to 1.97 L [42%]). This improvement was maintained during 21 months of follow-up. PMID- 11499829 TI - Digital medicine: the promise and the peril. PMID- 11499830 TI - Quinine-induced thrombocytopenia in a 64-year-old man who consumed tonic water to relieve nocturnal leg cramps. PMID- 11499831 TI - Primary esophageal motility disorders. PMID- 11499832 TI - Multiple aortic thrombi and protein C and S deficiency. PMID- 11499833 TI - C282Y and H63D mutation of the hemochromatosis gene in German porphyria cutanea tarda patients. AB - BACKGROUND AND AIMS: Patients with porphyria cutanea tarda (PCT) have a susceptibility to reversible inactivation of hepatocyte uroporphyrinogen decarboxylase, which can be triggered by alcohol, hepatitis C virus, and other agents. Inherited factors that may predispose to PCT include the C282Y mutation in the hemochromatosis (HFE) gene. METHODS: We analyzed the hemochromatosis mutations C282Y and H63D in liver biopsies and serum samples of 190 German patients (mean age 48+/-12.5 years) with sporadic PCT. The hepatic iron concentration was determined within the liver tissue. Age-matched healthy blood donors (115 donors) served as controls. RESULTS: The C282Y and H63D mutations were found in 75 (39%) and 85 (45%) of 190 patients with PCT, respectively. Twenty-two patients (12%) were homozygous for the C282Y mutation, and eighteen patients (9%) were compound heterozygotes, displaying both the C282Y and the H63D mutation. Within the control group, 3 of 115 patients were heterozygous for C282Y (3%) and 12 for H63D (10%). Serum and hepatic iron, ferritin, transferrin saturation, or liver enzymes did not differ significantly between patients with or without HFE mutations. CONCLUSIONS: The high frequency of homo- and heterozygosity for the C282Y and H63D alleles strongly suggests that these mutations are important predisposing factors for PCT in German patients. PMID- 11499834 TI - Are mammary pagetoid foam cells histiocytic or epithelial? PMID- 11499835 TI - Identification of Paneth cells in pyloric glands associated with gastric and intestinal mixed-type intestinal metaplasia of the human stomach. AB - We have proposed that intestinal metaplasia (IM) of the human stomach be divided into two types on the basis of cell differentiation status: a gastric and intestinal (GI) mixed type and a solely intestinal (I) type. In the GI mixed type, gastric (foveolar epithelial and pyloric gland cells) and intestinal (goblet, intestinal absorptive, and Paneth cells) phenotype cells coexist in the same intestinalized gastric glands in various combinations and degrees. Consequently, intestinalized gastric glands are hybrids. Although we have described the rare appearance of Paneth-like cells in pyloric glands of GI mixed type IM, the absence of an appropriate Paneth cell marker leaves room for doubt as to their true character. The purpose of this study was to clearly identify Paneth cells in pyloric glands in IM lesions using a new Paneth cell marker, a polyclonal antibody human defensin (HD)-5, raised against HD-5, which is included in granules of Paneth cells. A total of 105 gastric samples (4 biopsy and 101 surgical resected specimens) were examined. In only nine cases (8.6%), the antibody allowed demonstration of Paneth cells in pyloric glands in GI mixed-type IM, confirming our previous finding. Analysis of the proliferative cell (P) zone indicated that a common stem cell might generate both GI phenotype cells by upward and downward migration. No Paneth cells were found above the P zone. The results suggest that the stem cells show abnormal cell differentiation in IM lesions but preserve their normal direction of migration. PMID- 11499836 TI - Human oesophagostomiasis: a histomorphometric study of 13 new cases in northern Ghana. AB - Oesophagostomiasis is an infrequently described and recognised parasitic infection in humans, caused by Oesophagostomum bifurcum. Although the disease is most often found in the northern part of Togo and the neighbouring part of Ghana, sporadic cases have been described in other parts of Africa and in Asia and South America: Uganda, Ivory Coast, Sudan, Kenya, Ethiopia, Indonesia, Malaysia and Brazil. Infection probably occurs by way of the ingestion of L3 larvae. These larvae penetrate the intestinal wall, especially that of the colon. Some of these larvae develop into young adult worms and return to the bowel lumen. Other larvae, however, develop into immature worms, which fail to settle in the lumen, forming abscesses in the bowel wall and causing pathology. In the literature 105 human cases have been described, many originating in the northern regions of Ghana and Togo. The present study was performed to evaluate 13 new cases originating in the northern part of Ghana (7 female and 6 male patients, aged between 2 and 60 years). Histopathologically, the patients could be divided into two groups: the first group showed multinodular disease, while patients in the second group presented with a single, nodular mass. In the first group, abscesses were seen throughout the colonic wall. The mean size of the cavities was 4.3+/ 0.7 mm. There was no relation between the size and the localisation in the colonic wall. Abscesses were significantly larger in male patients than in female patients. There was no correlation with age. In the second group, histopathological examination showed a cyst of variable wall thickness with very limited inflammation. These cysts represented older lesions, often encapsulated in the mesentery. In conclusion, in this study we present 13 new cases of human oesophagostomiasis. The abscess formation was found to be organ specific, independent of age, and gender-related, producing a more intense tissue reaction in male patients. PMID- 11499837 TI - Renal expression of intercellular adhesion molecule-1 in immunoglobulin A nephropathy: tubulointerstitial injury and prognosis. AB - In immunoglobulin A nephropathy (IgAN), the abnormal expression of intercellular adhesion molecule-1 (ICAM-1) on proximal tubule epithelium is associated with the glomerular and interstitial infiltration of leucocytes, but its clinical significance remains uncertain. We analysed the relationship between the ICAM-1 (CD54) expression in tubular epithelial cells and interstitial leucocytes, macrophages (CD14) and T lymphocytes (CD3) with the histologic features, proteinuria and serum creatinine at the time of renal biopsy and after 2.42 years in 45 patients with IgAN and after 1.8+/-1.5 years in 29 patients with non glomerulonephritis (non-GN). In IgAN, ICAM-1+ tubule epithelium was 0.1+/-0.18 (x+/-SD), and this was associated with extracapillary proliferation (up to 20% of Bowman's space), glomerular sclerosis involving less than 50% of glomerular area, interstitial cellular infiltration, tubular atrophy and proteinuria level. ICAM 1+ interstitial leucocytes were correlated with glomerular sclerosis involving less than 50% of glomerular area, glomerular sclerosis involving more than 50% of glomerular area, tubular atrophy, interstitial fibrosis and serum creatinine level. In patients with an increase of 50% in serum creatinine, ICAM-1+, CD14+ and CD3+, interstitial leucocytes were significantly outnumbered than in patients with stable serum creatinine. In non-GN, ICAM-1+ tubule epithelium was 0.02+/ 0.04 (U=344, P<0.05, vs IgAN), and this was inversely correlated with the percentage of the normal glomeruli and associated with glomerular sclerosis covering more than 50% of glomerular area, tubular atrophy and serum creatinine level. The association between tubular ICAM-1 and proteinuria and the association between interstitial ICAM-1+, CD14+ and CD3+, leucocytes and renal failure at presentation and the deterioration in IgAN in contrast with non-GN suggest that tubular and interstitial expression of ICAM-1 may be a marker of tubulointerstitial disturbance in IgAN. PMID- 11499838 TI - Cytokines and cell cycle regulation in the fibrous progression of crescent formation in antiglomerular basement membrane nephritis of WKY rats. AB - Cytokines may regulate cell proliferation by cell-cycle-regulatory proteins, in which cyclin-dependent kinase inhibitors (CDKI) inhibit cell proliferation. We investigated whether CDKI p21 or p27, both of which are potentially regulated by transforming growth factor (TGF)-beta, a key cytokine in fibrogenesis, are involved together with TGF-beta and/or platelet-derived growth factor (PDGF) in the fibrous progression of glomerular crescent formation and examined the sequential change in the cell type and the cellular background of myofibroblasts in crescent formation. Crescentic glomerulonephritis (GN) was induced by i.v. injection of rabbit antirat glomerular basement membrane antiserum in WKY rats. Animals were killed 1, 2, 3 and 4 weeks after the induction of GN, and their kidneys were processed for immunohistochemical examination. After 1 week more than 85% of glomeruli showed cellular crescents, which became fibrocellular with decreased cellularity by 4 weeks. ED 1-positive macrophages were components of crescent cells in about 44% at 1-2 weeks, and this proportion declined markedly afterwards. Alpha smooth muscle actin (alpha SMA, a marker for myofibroblasts) positive cells were found in Bowman's epithelial cells (BEP) and in some crescent cells at 1 week, becoming major components of crescent cells by 4 weeks (about 40%). It was 2 weeks before invasion of alpha SMA-positive interstitial cells into glomeruli was evident. PDGF-B and PDGF receptor beta-positive cells, indicating possible targets for PDGF, were found in BEP adjoining crescent formation almost exclusively from 1 to 2 weeks. By contrast, both TGF-beta receptor types I- and II-positive cells, indicating possible effectors for TGF beta, were found in BEP and crescent formation, and the percentage of these in the crescent formation did not change until 4 weeks (about 32%). Cells with positive immunostaining for proliferating cell nuclear antigen and cyclin A, markers for cell proliferation, in the crescent formation peaked in number and proportion at 1-2 weeks, then decreased. In contrast, cells with positive immunostaining for p21 and p27, CDKI, were sparse at 1 week, and then increased markedly in number and in proportion, peaking at 3 (39.6%) or 2-3 weeks (about 25 30%), respectively. The present study demonstrates that restrained expression or a transient increase in p21 and p27 may be associated with proliferation or with inhibited proliferation of crescent cells, most of which are macrophages and myofibroblasts. The action, of PDGF and TGF-beta may contribute to the recruitment of myofibroblasts into the crescent. The action of TGF-beta on crescent cells might be linked to the expression of p21 and/or p27. PMID- 11499839 TI - Enhanced in situ expression of aldose reductase in peripheral nerve and renal glomeruli in diabetic patients. AB - To explore the relationships between polyol pathway-related enzymes and pathologic features, we examined the immunohistochemical expression of aldose reductase (AR) and sorbitol dehydrogenase (SDH) in the peripheral nerve and kidney tissues collected postmortem from diabetic patients and compared it with those from non-diabetic patients. Tissue AR protein concentrations were also quantified. In non-diabetic patients, AR distributed in pericytes, smooth muscle cells of endo- and epi-neurial microvessels, Schwann cells in the sciatic nerve, and tubular cells of the renal medulla. By contrast, positive SDH reactions were observed in tubular cells of the renal cortex but were faint in the sciatic nerve. Diabetic patients frequently showed dense AR expressions in the sciatic nerve. In nephropathic diabetic patients, the glomerular mesangial area showed diffuse positive reactions for AR. The severity of structural changes in glomeruli correlated with the intensity of immunoreactive AR (r2=0.626, P<0.01). AR contents in the renal cortex and sciatic nerve from diabetic patients were 1.5 and 1.8-fold greater than those from non-diabetic patients, respectively (P<0.05 for both). These findings are the first to demonstrate enhanced AR expressions in peripheral nerve and renal glomeruli in diabetic patients and its relevance to the characteristic pathology. PMID- 11499840 TI - Expression of cyclin-dependent kinase inhibitors p16MTS1, p21WAF1, and p27KIP1 in HPV-positive and HPV-negative cervical adenocarcinomas. AB - Inactivation or down-regulation of the cell-cycle inhibitors p16MTS1, p21WAF1, and p27KIP1 is involved in the carcinogenesis of various human tumors. In cervical squamous cell carcinomas that are associated with human papillomavirus (HPV) infection, the expression or function of these proteins is impaired by the action of viral oncoproteins E6 and E7. Comparably less is known about the role of these cyclin-dependent kinase inhibitors in cervical adenocarcinomas, 15-40% of which are HPV negative. Therefore, we studied the expression of p16MTS1, p21WAF1, and p27KIP1 by immunohistochemistry in 60 cervical adenocarcinomas. HPV infection was determined by PCR, and HPV 16 and 18 E6/E7 oncogene expression was analyzed by RNA-RNA in situ hybridization. We found significant correlations of strong p16 expression with HPV 16/18 infection and HPV 16/18 E6/E7 oncogene expression (P=0.001). Moderate or strong p16 expression was also observed in 41% of HPV-negative carcinomas, indicating that HPV-independent mechanisms might also lead to p16 overexpression. In addition, stronger p21 and p27 expression was significantly associated with the detection of HPV 16 or 18 E6/E7 transcripts (P=0.015 and 0.030, respectively). Obviously, the tumor suppressor action of these proteins can be overcome in HPV-positive lesions. In contrast, absent or low p16, p21, and p27 immunostaining was observed in most HPV-negative cervical adenocarcinomas and might contribute to carcinogenesis in these tumors. PMID- 11499841 TI - Invasive carcinoma derived from intraductal papillary-mucinous carcinoma of the pancreas: clinicopathologic and immunohistochemical study of eight cases. AB - Most intraductal papillary-mucinous carcinomas (IPMCs) of the pancreas are resectable and curable, but some develop into frankly invasive carcinomas. We studied the clinicopathologic features of eight cases of invasive carcinoma derived from IPMC (IC-IPMC) of the pancreas. The patients were aged 54-75 years (mean, 66.6 years); six were male and two were female. The mean tumor size was 7.7 cm (range 5.5-10.5 cm). Two patients without lymph node metastasis had no peripancreatic invasion, and survived longer (115 and 20 months). Three out of four patients with extrapancreatic invasion died of their tumors or developed tumor recurrence within a year. One patient with evidence of liver and lymph node metastasis at the time of first surgery again showed metastatic tumor 21 months later. One patient died of another cause. We also performed a comparative study of the immunohistochemical features of IC-IPMCs in 9 IPMCs (including minimally invasive cases) and 15 ductal adenocarcinomas. CEA cytoplasmic positivity was observed in most of the IC-IPMCs (87.5%) and ductal adenocarcinomas (93.3%), but in only 1 IPMC (11.1%). The frequency of p53 nuclear staining in ductal adenocarcinoma (73.3%) was higher than in IPMC (33.3%) or IC-IPMC (37.5%). In conclusion, IC-IPMC with extrapancreatic invasion should be treated as ductal carcinoma because of its aggressive behavior after resection. Some IPMCs might progress to invasive carcinoma via pathways that are different from those followed by ductal adenocarcinomas. PMID- 11499842 TI - Estrogen receptor-alpha and C-ERBB-4 expression in breast carcinomas. AB - The presence of estrogen receptors (ERs) in breast carcinomas is important for clinical response to endocrine therapy. However, the cellular mechanisms following ER activation are not fully understood. It has been indicated that expression of the ER is associated with the expression of c-erbB-4. To address this question, 103 breast carcinoma samples were studied using reverse transcriptase polymerase chain reaction (RT-PCR) analysis after application of a microselection method for RNA isolation. Total RNA for RT-PCR was isolated from 20-microm-thick frozen sections, which were made from microselected areas. Paraffin blocks from 98 of these 103 tumors were also immunohistochemically examined. Significant associations between ER-alpha and c-erbB-4 mRNA and protein expressions were found in the present study with both methods. One-fourth of the tumors did not express ER-alpha (22%, 24%, and 26% with chemical binding, immunohistochemistry, and RT-PCR, respectively). About one-half of the ER-alpha negative tumors did not express c-erbB-4 on both mRNA and protein levels (48% with RT-PCR and 46% with immunohistochemistry, P=0.001 for both methods). The endocrine therapy responsive breast cancer cell lines MCF-7 and T47-D were positive for both ER-alpha and c-erbB-4 expression, while the endocrine therapy nonresponsive breast cancer cell lines MDA-MD-231 and SK-BR-3 were not. Thus, we confirm the association between the expression of ER-alpha and c-erbB-4 mRNA and protein in breast carcinomas, indicating a role for c-erbB-4 in estrogen signal transduction. PMID- 11499843 TI - Spindle cell ductal carcinoma in situ. An unusual variant of ductal intra epithelial neoplasia that simulates ductal hyperplasia or a myoepithelial proliferation. AB - Seventeen examples of a variant of ductal carcinoma in situ (DCIS) composed exclusively or predominantly of spindle cells arranged in fascicles, whorls, and solid sheets are described. The fascicular arrangement of the spindle cells simulates the "streaming" phenomenon associated with ordinary intraductal epithelial hyperplasia (IDH). This process also resembles the myoid, solid form of intraductal myoepithelial proliferation. The women ranged in age from 38 years to 79 years with a mean age of 59.3 years. Five patients presented with a palpable mass. The remaining tumors were discovered using mammography. The radiological appearances of the lesions raised concern for carcinoma, but there were no distinctive mammographic findings to suggest an unusual variant of DCIS. Cytological preparations were suspicious for malignancy in two patients and were reported as malignant in another case. Sixteen patients were treated with wide local excision, and one woman had a partial mastectomy. The tumors measured from 3 mm to 15 mm (mean 8.65 mm). In three cases, minute foci of stromal invasion were associated with the spindle cell DCIS. In another specimen, a 2.7-cm invasive ductal carcinoma of no special type was identified in an area away from the foci of the spindle cell DCIS. None of the patients has experienced recurrence or metastasis during the relatively short mean follow-up period of 16.2 months (range 4-77 months). Spindle cell DCIS is distinguished from the streaming pattern of ordinary IDH by its solid growth pattern, lack of secondary spaces or peripheral fenestrations, uniformity of appearance and distribution of nuclei, cytological atypia in the range of low to intermediate-grade DCIS, and negative immunoreaction with CK-34betaE12 (HMW-CK903). When fenestrations are present, they are evident in areas of cribriform DCIS that merge with the solid, spindle cell areas in hybrid ducts harboring both patterns. This admixture, with conventional cribriform DCIS, and the association with foci of invasive ductal carcinoma in some cases further help recognition and confirmation of this lesion as in situ carcinoma. When there is no transition from the spindle cells to recognizable cribriform DCIS, distinction from intraductal myoepithelial hyperplasia (myoepitheliosis) requires immunostains for actin and S-100 protein. Recognition of this pattern of DCIS is important in order to avoid its frequent misclassification as a benign lesion. PMID- 11499844 TI - Constrictive bronchiolitis obliterans. Characterisation of fibrogenesis and lysyl oxidase expression patterns. AB - The process leading to irreversible fibrotic constriction of the bronchioles was studied in two cases of bronchiolitis obliterans (BO) after bone marrow transplantation. Because lysyl oxidase (LOX) is the main collagen cross-linking enzyme that might account for irreversible fibrosis, its expression was studied together with expression of extracellular matrix (ECM) proteins. Characteristic types of lesions could be distinguished on the basis of histological and immunohistological criteria. An inflammatory stage was characterised by infiltration restricted to the bronchioles by lymphocytes and dendritic cells. A fibro-inflammatory stage was characterised by the coexistence of a persistent immune cellular lesion pattern with further focal modelling of a sub-epithelial neo-synthesised connective matrix. LOX expression was observed at the tips of intra-luminal fibrotic protrusions, together with tenascin and cellular fibronectin. A fibrotic stage was characterised by dense ECM deposits spreading throughout the peri-bronchiolar connective tissue, resulting in bronchiole obliteration and final disappearance. In contrast to reversible cases of fibrosis, persistence of long-term LOX expression reflecting continuing fibrosing activity might account for the irreversible status of BO. Our two cases illustrated that, at inflammatory and fibro-inflammatory stages, BO may be stabilised by immunosuppressive treatment, while the persistence of LOX expression in the fibrotic stage might correspond to a disease that becomes irreversible and fatal. PMID- 11499845 TI - Molecular cytogenetic analysis of two primary squamous cell carcinomas of the lung using multicolor fluorescence in situ hybridization. AB - To fully characterize the numerous chromosomal aberrations in two human squamous cell carcinomas (SCCs) of the lung, molecular cytogenetic characterization was carried out utilizing conventional banding analysis and multicolor fluorescence in situ hybridization (mFISH), providing simultaneous color discrimination of all 24 human chromosomes. Both tumors displayed complex aneuploid karyotypes with a host of numerical and structural chromosome abnormalities. Structural aberrations common to both SCCs included rearrangements of chromosomes 1, 3p, 7q, and 8q, contributing to net loss of chromosomal sequences on 1p, 3p, and 8p, and a net gain of 8q. The recently introduced mFISH technique enabled the disclosure of cryptic translocations and the chromosomal composition of previously unrecognized marker chromosomes. Furthermore, mFISH greatly enhanced the ability to delineate chromosomal breakpoints when integrating banding information from conventional banding analysis. Eventually, the application of mFISH as a powerful approach to refine complex tumor karyotypes is expected to result in a more detailed and complete picture of cytogenetic events associated with the development and progression of solid tumors. PMID- 11499846 TI - Sulfatide storage in visceral organs of arylsulfatase A-deficient mice. AB - The inherited deficiency of arylsulfatase A (ASA) in humans causes lysosomal accumulation of sulfatides in visceral organs and in the nervous system and leads to wide-spread demyelination (metachromatic leukodystrophy, MLD). ASA-deficient mice have previously been generated by means of targeted gene disruption. In the present study, visceral organs of ASA-deficient mice were investigated. A simple technique for the histochemical detection of accumulated sulfatides was elaborated using pre-embedding staining with alcian blue. The gall bladder, intrahepatic bile ducts, exocrine pancreatic ducts, respiratory epithelium and, with low degree, testicular Sertoli cells, showed sulfolipid storage. The storage pattern in the kidney will be described in a separate publication. Hepatocytes, pancreatic islets, adrenal glands, and gastric epithelium were unaffected. Ultrastructurally, the intralysosomal storage material displayed parallel and concentric lamellar patterns. Apart from some differences, the topographic distribution of the sulfatide storage resembled that in human MLD. In addition to being an animal model of the human disease, the ASA-deficient mouse may be useful for investigating the cell biology of sulfolipids in visceral organs. PMID- 11499847 TI - Uterine tumor resembling ovarian sex-cord tumor--a case report and review of the literature. AB - A uterine tumor resembling an ovarian sex-cord tumor (UTROSCT) is a very rare lesion with only 38 cases reported in the literature so far. Here, we show an additional case of a pure UTROSCT with a DNA stemline at 1c in a 49-year-old woman presenting with abnormal vaginal bleeding. Problems in differential diagnosis arise mainly due to the variable histological picture of UTROSCT. Immunohistochemically, these tumors express cytokeratin, epithelial membrane antigen, vimentin, and smooth muscle actin. Moreover, in some cases, CD99 and alpha-inhibin are detectable. Although 36% of UTROSCT have infiltrative margins, almost all of them behave benignly. It is thus questionable whether the same prognostic criteria apply for these tumors as for endometrial stromal sarcomas. However, in the so-called mixed UTROSCT, the endometrial stromal sarcoma component determines the outcome. PMID- 11499848 TI - The multiple propensity score for analysis of dose-response relationships in drug safety studies. AB - In order to detect adverse drug reactions, large observational drug safety studies are necessary as randomized clinical trials rarely have enough power. However, in order to obtain reliable results the issue of confounding, especially confounding by indication, should be addressed. We proposed a multiple propensity score, which is an extension of the propensity score, to reduce the bias in a dose-response analysis in a drug safety study. The multiple propensity score has similar properties to the propensity score in Rosenbaum and Rubin.(1) Using the propensity score for bias reduction of the risk ratio was considered. We used the multiple propensity score in a study of the dose-response relationship between diclofenac prescriptions and hospitalization for gastrointestinal bleeding and perforation using a record linkage database. We found that the unadjusted risk ratios were biased downwards due to confounders and that this bias was reduced by using stratification based on the multiple propensity score. PMID- 11499849 TI - Identifying hospital admissions due to adverse drug events using a computer-based monitor. AB - BACKGROUND: Hospital admissions due to adverse drug events (ADEs) are expensive, and many may be preventable, yet few institutions have ongoing surveillance for these events. OBJECTIVE: To evaluate the use of a computer-based ADE monitor to identify admissions due to ADEs and to measure the associated costs. DESIGN: Prospective cohort study in one tertiary care hospital. PARTICIPANTS: All patients admitted to nine medical and surgical units in a tertiary care hospital over an 8-month period. MAIN OUTCOME MEASURE: Admissions to the hospital due to an adverse drug event. METHODS: A computer-based monitoring program generated alerts suggesting that an ADE might be present. A trained reviewer then evaluated the record. RESULTS: Among the 3238 admissions, 76 (2.3%, 1.4% after adjusting for sampling) were found to be caused by an ADE. Of these ADEs, 78% were severe and 28% were preventable. Estimated costs were $16,177 per ADE, and $10,375 per preventable ADE; annualized costs to the hospital were $6.3 million per year for all ADEs, and $1.2 million for preventable ADEs. CONCLUSIONS: Many admissions were caused by ADEs, although our point estimate undoubtedly represents a lower bound. These events were mostly severe, often preventable, and expensive. The computer-based monitoring system represents a practical approach for identifying ADEs that occur in outpatients and cause admission to the hospital. PMID- 11499850 TI - Adult height in women with childhood asthma--a population-based study. AB - PURPOSE: To study adult height in children that grew up with asthma before inhaled steroids became first-line therapy. METHODS: Data from the Swedish Medical Birth Register (self-reported asthma) and the Hospital Discharge Register (first hospitalization for asthma) were used, to compare adult height for asthmatic and non-asthmatic pregnant women. The analysis was restricted to women in their first full-term pregnancy, born in Sweden between 1960-1974 and of Swedish citizenship. RESULTS: The mean height of all the women in the study population was 166.7 cm (SD = 8.8, n = 287,750) and of the women who reported asthma 166.5 cm (SD = 6.1, n = 13,059, p < 0.01). The mean height of women first hospitalized because of asthma at age 0-8 years was 165.5 cm (SD = 5.9 cm, n = 555, p < 0.01). Among the asthmatic women, there was no skew distribution of heights. CONCLUSIONS: Girls with moderate to severe childhood asthma who grew up before inhaled glucocorticosteroids became first-line therapy attained 0.7-1.2 cm lower adult height. The differences in height. while of statistical relevance, are not clinically relevant. PMID- 11499851 TI - Population screening for isoniazid acetylator phenotype. AB - PURPOSE: To establish a useful method for acetylator phenotypification and therapeutic drug monitoring of patients receiving isoniazid. METHODS: Sixty patients with uncomplicated pulmonary tuberculosis were given a 5-mg/kg oral dose of isoniazid each. Plasma concentrations of isoniazid and its metabolite, acetyl isoniazid, were determined by HPLC analyses at various post-dose times. From the isoniazid concentration and the concentration ratio of acetyl-isoniazid and isoniazid (metabolic ratio), phenotypification methods were assessed. RESULTS: The metabolic ratios at 3 h post-dose revealed a trimodal distribution; a fast, intermediate and slow acetylator phenotype group. The 2-h and 6-h data showed different bimodal combinations of these phenotype groups. The metabolic ratio phenotypification method could be simplified by using the HPLC data directly without converting it to absolute concentrations. CONCLUSIONS: A single-sample test based upon the plasma isoniazid concentration, combined with the metabolic ratio of acetyl-isoniazid and isoniazid, appears to be a reliable parameter for phenotype discrimination and for bioavailability testing. PMID- 11499852 TI - Association between terbinafine and arthralgia, fever and urticaria: symptoms or syndrome? AB - PURPOSE: The antifungal agent terbinafine has been approved for marketing in The Netherlands since 1992. Adverse drug reactions (ADRs) may occur in about 10% of the patients, the majority gastrointestinal disorders and skin reactions. Since the introduction of terbinafine, the Netherlands Pharmacovigilance Foundation Lareb received eight reports of arthralgia during the use of this drug. In four reports the additional presence of skin reactions was mentioned, two of these reports concerned urticaria. Two patients who reported arthralgia also had a fever. These reports were described in more detail, and analysed statistically in order to determine whether symptoms are interrelated. METHODS: All reports with known gender and a reporting date between 1 March 1992 and 1 January 1999, concerning patients older than 10 years, were included. The extent to which the symptoms urticaria, fever and arthralgia were interrelated was examined by logistic regression modelling. RESULTS: Case series as well as the results of the statistical analysis show a clustering of symptoms among reports of patients using terbinafine. Both urticaria and arthralgia were statistically significantly associated with reports on terbinafine compared to all other reports in the database. CONCLUSION: The findings might point towards a clustering of these symptoms in patients using terbinafine. Possibly these symptoms have a shared aetiology, presumably an immunological reaction. PMID- 11499853 TI - Minimized analysis of costs applied to antimicrobial consumption in a rural area. AB - PURPOSE: Antimicrobial agents constitute one of the most utilized groups of drugs in daily clinical practice and, therefore they involve a significant expense. The aim of this study was to evaluate the economic cost of the antimicrobials prescribed in a rural area as well as to search for some cheaper alternatives. METHODS: Retrospective study. The economic cost of antimicrobial agents prescribed at a health centre over 18 months was studied. To do this, clinical histories of 800 people were reviewed. Afterwards, a minimized analysis of costs was carried out. RESULTS: The total cost of antimicrobial consumption came to 2,080.752 pts. The average expenditure per patient came to 6,433.85 +/- 14 269.29 pts. Significant differences between the sexes were not found; however, the expenditure in patients of 65 years of age or over was significantly higher than the rest. After applying the ABC analysis it was noticed that macrolides, cephalosporins, antimicrobial combinations and quinolones were the most important groups from an economic point of view. The use of monodose containers would allow us to save up to 7.83% of the total expenditure. In addition, by prescribing the cheapest marketed pharmaceutical product we could save a further 6.54%, and, finally, by combining these two measures the total possible saving would reach 299,052 pts, a 14.37% of the total expenditure. CONCLUSION: We consider important the elaboration of pharmacoeconomic guides as well as the introduction of monodose containers not only at hospitals but also at community pharmacists. PMID- 11499854 TI - Survey of knowledge and practice on oral contraceptive and emergency contraceptive pills of drugstore personnel in Hat Yai, Thailand. AB - In Thailand, oral contraceptive (OC) and emergency contraceptive pill (ECP) are available as over-the-counter (OTC) drugs, and drugstores share 30% of services. While the rate of dispensing contraceptive pills has increased, the knowledge and awareness of ECP use is limited among users and providers. The objective of this study was to assess knowledge and practice of drugstore personnel on providing OC and ECP, in order to improve the quality of services. Drugstores located in Hat Yai District, Songkhla Province, Southern Thailand, were the accessible population. There were 109 drugstores, half of them owned by pharmacists. The population was stratified by owner (pharmacist or non-pharmacist) and randomly selected to obtain a sample size of 30 drugstores for each class. Two study methods, questionnaire interview and secret shopping, were used to measure knowledge, and practice, respectively. History-taking, drug-choosing, and advice giving were the domains measured. The results demonstrated that knowledge on OC was fair, but that on ECP was poor. Pharmacists had better knowledge of proper history taking and ECP indication than non-pharmacists. OC and ECP provision were inappropriately practised in drugstores in the study area. A majority of drugstores were mainly owned by non-pharmacists. For OC practice, drug-choosing was good, but history-taking and advice-giving were poor in both groups. Although both groups dispensed ECP poorly, pharmacists dispensed significantly better than non-pharmacists. Among non-pharmacist staff, the average scores of OC advice giving, and ECP dispensing, were statistically significantly better among those working in pharmacist-owned drugstores. Both knowledge and practice on OC and ECP should be improved in both types of drugstores in the study area. PMID- 11499855 TI - The risk of obstructive airways disease in a glaucoma population. AB - BACKGROUND: Asthma and chronic obstructive pulmonary disease (COPD), defined as obstructive airways disease (OAD), are two common chronic conditions especially in the elderly. Glaucoma is also a common disease in the elderly with a prevalence close to 5% among those older than 75 years. Most medical therapy for glaucoma is given as eye drops. It has been described that small amounts of systemically absorbed beta-blockers can produce significant respiratory adverse events in predisposed patients. METHODS: Population-based cohort study with nested case-control analysis using the UK General Practice Research Database (GPRD). We studied the prevalence of OAD in a cohort of patients 60 to 85 years old with a first ever diagnosis of glaucoma and compared it to the prevalence in an age- and sex-matched cohort sampled from the general population. We also calculated the RR and 95% CI of worsening asthma in non-severe asthma patients among the two cohorts. Incidence of OAD was studied in a cohort of glaucoma patients 60 to 85 years old and in an age- and sex-matched cohort from the general population. RESULTS: The prevalence of OAD was the same between the glaucoma cohort and the general population (OR 1.1; 95% CI 0.9-1.4). The risk of worsening asthma associated with glaucoma was OR 1.2 (95% CI 0.5-2.8). The incidence of OAD was similar in both cohorts. Current users of ophthalmic drugs presented a RR of 1.2 (95% CI 0.8-1.9) of developing asthma compared to non-users in the glaucoma population. The risk in the first month of treatment with topical beta-blockers was 2.1 (95% CI 0.7-6.7). DISCUSSION: We did not find an association between glaucoma and OAD. Use of topical glaucoma medication was not associated with a major increased risk of asthma. PMID- 11499856 TI - Pharmacoepidemiology--a tool for public health. PMID- 11499857 TI - Pharmacoepidemiology and drug safety. PMID- 11499858 TI - Methylphenidate in children with hyperactivity: review and cost-utility analysis. AB - OBJECTIVE: To evaluate the effectiveness and cost effectiveness of methylphenidate in the treatment of children with hyperkinetic disorder as defined using ICD-10 criteria. DESIGN: Comprehensive literature review and cost utility analysis comparing methylphenidate treatment with placebo. Costs and effects were estimated from a NHS perspective according to the methodology developed by the previous South and West Development and Evaluation Committee. The number of Quality Adjusted Life Years (QALYs) gained was estimated by using the Index of Health Related Quality of Life to model treatment effects. RESULTS: Evidence from good and medium quality randomized controlled trials shows benefits of methylphenidate over weeks and months respectively. Evidence beyond 6 months is poorer and it is uncertain whether effects of methylphenidate persist into adolescence and adulthood. Methylphenidate is of reasonable cost-effectiveness when considering short- and medium- term benefits with an estimated cost per QALY of 7 pounds 400 to 9 pounds 200 at 1997 prices. CONCLUSIONS: Short-term treatment of hyperkinetic children with methylphenidate is effective and cost effective. PMID- 11499859 TI - Prospective cohort study of adverse events monitored by hospital pharmacists. Hospital Adverse Event Monitoring Study (HAEMS) Group. AB - PURPOSE: To examine the feasibility of pharmacist-led intensive hospital monitoring of adverse events (AEs) associated with newly marketed drugs. SUBJECTS/SETTING: 303 patients admitted to Southampton Hospitals who were prescribed selected newly marketed drugs during their inpatient stay in 1998. METHODS: Prospective observational study. Patients were identified from computerized pharmacy records, clinical pharmacist ward rounds, dispensary records or via nursing staff. The pharmacist reviewed medical notes and recorded AEs, suspected adverse drug reactions (ADRs) and reasons for stopping drugs. OUTCOMES: Incidence of AEs, ADRs; proportionate agreement between the physician's and pharmacist's event recording. RESULTS: 303 patients were monitored. Of the patients taking newly marketed drugs 92% were identifiable using pharmacy computer systems and pharmacist ward visits. There were 21 (7%) suspected ADRs detected during this pilot study. The types of adverse events detected were broadly similar to those identified by general practice-based prescription event monitoring. However, biochemical changes featured more frequently than in general practice. Differences between adverse events recorded by pharmacist and physician were systematic and attributed to differences in event coding. CONCLUSION: Pharmacist-led monitoring in a typical NHS hospital setting was effective at detecting ADRs in newly marketed drugs. However, this effort might have been substantially less time-consuming and more effective were electronic patient records (EPRs) available. Pharmacy computer systems are not designed to be patient focused and are therefore unable to identify patients taking newly marketed drugs. It is argued that future EPR and computerised patient-specific prescribing systems should be designed to capture this data in the same way as some US systems are currently able to do. PMID- 11499860 TI - A clinical approach to diagnosis and management of Paget's disease of bone. PMID- 11499861 TI - Female estrogen receptor beta-/- mice are partially protected against age-related trabecular bone loss. AB - Recently, it has been shown that inactivation of estrogen receptor beta (ER-beta) by gene targeting results in increased cortical bone formation in adolescent female mice. To study the possible involvement of ER-beta in the regulation of the mature skeleton, we have extended the analyses to include 1-year-old ER-beta knockout mice (ER-beta-/-). Male ER-beta-/- mice did not express any significant bone phenotypic alterations at this developmental stage. However, the increase in cortical bone parameters seen already in the adolescent female ER-beta-/- mice was maintained in the older females. The aged female ER-beta-/- mice further exhibited a significantly higher trabecular bone mineral density (BMD) as well as increased bone volume/total volume (BV/TV) compared with wild-type (wt) mice. This was caused by a less pronounced loss of trabecular bone during adulthood in female ER-beta-/- mice. The growth plate width was unaltered in the female ER beta-/- mice. Judged by the expression of the osteoclast marker tartrate resistant acid phosphatase (TRAP) and cathepsin K (cat K; reverse-transcription polymerase chain reaction [RT-PCR]) as well as the serum levels of C-terminal type I collagen cross-linked peptide, bone resorption appeared unaffected. However, an increase in the messenger RNA (mRNA) expression levels of the osteoblast marker core-binding factor alpha1 (Cbfa1) suggested an anabolic effect in bones of old female ER-beta-/- mice. In addition, the mRNA expression of ER alpha was augmented, indicating a role for ER-alpha in the development of this phenotype. Taken together, the results show that ER-beta is involved in the regulation of trabecular bone during adulthood in female mice and suggest that ER beta acts in a repressive manner, possibly by counteracting the stimulatory action of ER-alpha on bone formation. PMID- 11499862 TI - Sequential treatment with basic fibroblast growth factor and parathyroid hormone restores lost cancellous bone mass and strength in the proximal tibia of aged ovariectomized rats. AB - This study was designed to determine whether sequential treatment with basic fibroblast growth factor (bFGF) and parathyroid hormone (PTH) can restore lost cancellous bone mass and strength at a severely osteopenic skeletal site in aged ovariectomized (OVX) rats. Female Sprague-Dawley rats were subjected to sham surgery or ovariectomy at 3 months of age and maintained untreated for the first year after surgery. At 15 months of age, groups of baseline control and OVX rats were killed and catheters were inserted in the jugular veins of all remaining rats. Two groups of OVX rats were injected intravenously (iv) daily with bFGF for 14 days at a dose of 200 microg/kg body weight. At the end of bFGF treatment, one group was killed whereas the other group was subjected to 8 weeks of treatment with synthetic human PTH 1-34 [hPTH(1-34)] consisting of subcutaneous (sc) injections 5 days/week at a dose of 80 microg/kg. Another group of OVX rats was treated iv with vehicle for 2 weeks followed by treatment with PTH alone for 8 weeks. Other groups of sham-operated control rats and OVX rats were treated iv and sc with vehicle alone. The right proximal tibia from each rat was processed undecalcified for quantitative bone histomorphometry and the left proximal tibia was subjected to biomechanical testing. Baseline and vehicle-treated OVX rats were severely osteopenic because their tibial cancellous bone volumes were less than 5% compared with mean values of 20.3% and 15.0% in baseline and vehicle treated control rats, respectively. Treatment of OVX rats for 2 weeks with bFGF alone did not significantly increase tibial cancellous bone volume but induced marked increases in osteoid volume, osteoblast surface, and osteoid surface. Sequential treatment of aged OVX rats with bFGF and PTH increased tibial cancellous bone volume (15.1%) and load to failure to at least the level of vehicle-treated control rats. Tibial cancellous bone volume (10.8%) and load to failure also were significantly increased by treatment with PTH alone, and these variables were not significantly different from those of OVX rats treated with bFGF + PTH. However, tibial ash density was significantly greater in OVX rats treated sequentially with bFGF and PTH compared with OVX rats treated with PTH alone. Our findings suggest that sequential treatment with bFGF and PTH may be useful for restoration of lost cancellous bone in the severely osteopenic, estrogen-deplete skeleton, but it cannot be concluded with certainty that this sequential treatment has a greater bone restorative effect than treatment with PTH alone. PMID- 11499863 TI - Relation between vitamin D insufficiency, bone density, and bone metabolism in healthy postmenopausal women. AB - Although only few postmenopausal women exhibit biochemical signs of hypovitaminosis D, vitamin D insufficiency has been shown to have adverse effects on bone metabolism and could be an important risk factor for osteoporosis and fracture. We determined serum levels of 25-hydroxyvitamin D [25(OH)D], intact parathyroid hormone (iPTH), bone turnover markers, dietary calcium intake, and bone mineral density (BMD; measured by dual X-ray absorptiometry) in 161 consecutive ambulatory women, healthy except for osteoporosis, referred to a bone metabolic unit. The prevalence of vitamin D insufficiency [25(OH)D < or = 15 ng/ml] was 39.1%. 25(OH)D was lower in the osteoporotic subjects (15.7 +/- 5.3 ng/ml vs. 21.8 +/- 9.7 ng/ml; p < 0.001). After controlling for all other variables, lumbar spine (LS) BMD was found to be significantly associated with 25(OH)D, body mass index (BMI), and years after menopause (YSM) (R2 = 0.253; p < 0.001). For femoral neck (FN), significant independent predictors of BMD were YSM, BMI, iPTH, and 25(OH)D (R2 = 0.368; p < 0.001). The probability of meeting osteoporosis densitometric criteria was higher in the vitamin D insufficiency group (odds ratio [OR], 4.17, 1.83-9.48) after adjusting by YSM, BMI, iPTH, and dietary calcium intake. Our study shows that vitamin D insufficiency in an otherwise healthy postmenopausal population is a common risk factor for osteoporosis associated with increased bone remodeling and low bone mass. PMID- 11499864 TI - Expression profiles of receptor activator of nuclear factor kappaB ligand, receptor activator of nuclear factor kappaB, and osteoprotegerin messenger RNA in aged and ovariectomized rat bones. AB - The receptor activator of nuclear factor-kappaB ligand (RANKL; also known as tumor necrosis factor-related activation-induced cytokine [TRANCE], osteoprotegerin ligand [OPGL], and osteoclast differentiation factor [ODF]) is a transmembrane ligand expressed in osteoblasts and bone marrow stromal cells. It binds to RANK, which is expressed in osteoclast progenitor cells, and induces osteoclastogenesis. OPG, a decoy receptor for RANKL, also binds to RANKL, and competitive binding of RANKL with RANK or OPG is thought to regulate bone metabolism. To investigate roles of the RANKL/RANK/OPG system in pathophysiological conditions, the expression of RANKL, RANK, and OPG messenger RNA (mRNA) was analyzed in bones of aged and ovariectomized rats by means of in situ hybridization. In the control 8-week-old male and sham-operated female rat bones, the expression of RANKL mRNA was detected in hypertrophic chondrocytes of the growth plate and some periosteal and endosteal mesenchymal cells. The expression of RANK mRNA was detected in osteoclast-like cells and mononuclear cells in contact with the cortical and trabecular bones. The expression of OPG mRNA was detected in proliferating chondrocytes and osteocytes. In the 2.5-year old rat bones, the expression of RANKL, RANK, and OPG mRNA tended to decrease except for the endosteal region. In the ovariectomized rat bones, the expression of RANKL, RANK, and OPG mRNA increased, and high expression of OPG mRNA was induced in resting chondrocytes and osteocytes. These results suggest that estrogen deficiency stimulates the RANKL/RANK/OPG system and induces OPG in cells that have been thought to be less important for bone metabolism. PMID- 11499865 TI - Leptin is expressed in and secreted from primary cultures of human osteoblasts and promotes bone mineralization. AB - The adipose hormone leptin and its receptor are important for regulation of food intake and energy metabolism. Leptin also is involved in the growth of different tissues. In this study, we show the expression of leptin in primary cultures of normal human osteoblasts (hOBs) as evidenced by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry. Release of leptin into the medium also was found. Leptin was not detected in commercially available hOBs (NHOst) or in three different human monoclonal osteosarcoma cell lines. Leptin expression was observed in OBs in the mineralization and/or the osteocyte transition period but not during the matrix maturation period. Furthermore, hOBs and osteosarcoma cell lines expressed the long signal-transducing form of the leptin receptor (OB-Rb) as shown by RT-PCR. We observed no significant changes in leptin or OB-Rb genes in hOBs after incubation with recombinant leptin, indicating no autoregulation of the leptin expression. Incubation of both hOBs entering the mineralization phase and osteosarcoma cell lines with recombinant leptin markedly increased the number of mineralized nodules as shown by alizarin S staining. These findings indicate that leptin may be of importance for osteoblastic cell growth and bone mineralization. PMID- 11499866 TI - Inhibition of Na+/Ca2+ exchange with KB-R7943 or bepridil diminished mineral deposition by osteoblasts. AB - Osteoblasts form new bone by secreting a complex extracellular matrix that has the capacity to mineralize when adequate amounts of calcium and phosphate are supplied. The studies reported here show that long-term treatment of cultured, primary osteoblasts with Na+/Ca2+ exchanger (NCX) inhibitors, bepridil and KB R7943, impacts in a dose-dependent manner the ability of the cells to form a calcified matrix. Treatment of confluent osteoblast cultures for 14 days with low levels of bepridil (3.0 microM) or KB-R7943 (1.0 microM and 0.1 microM) resulted in a significantly diminished capacity of these cells to mineralize bone matrix, without significantly altering cell morphology, viability, or cell differentiation. The data indicate that inhibition of NCX reduces mineral accumulation in the bone matrix by blocking the efflux of Ca2+ from the osteoblast into the bone fluid. In addition, immunocytochemistry of type I collagen (COLI) and bone sialoprotein (BSP) suggests that inhibition of NCX by 1.0 microM KB-R7943 also may impair the secretion of bone matrix proteins by the osteoblasts. This study is the first to show that NCX is an important regulator of the bone fluid microenvironment and that NCX appears critical to the mineralization process. PMID- 11499867 TI - Accelerated turnover of metaphyseal trabecular bone in mice overexpressing cathepsin K. AB - This study is based on a hypothesis that overexpression of an osteoclast enzyme, cathepsin K, causes an imbalance in bone remodeling toward bone loss. The hypothesis was tested in transgenic (TG) mice harboring additional copies of the murine cathepsin K gene (Ctsk) identifiable by a silent mutation engineered into the construct. For this study, three TG mouse lines harboring 3-25 copies of the transgene were selected. Tissue specificity of transgene expression was determined by Northern analysis, which revealed up to 6-fold increases in the levels of cathepsin K messenger RNA (mRNA) in calvarial and long bone samples of the three TG lines. No changes were seen in the mRNA levels of other osteoclast enzymes, indicating that the increase in cathepsin K mRNA was not a reflection of activation of all osteoclast enzymes. Immunohistochemistry confirmed that cathepsin K expression in the TG mice was confined to osteoclasts and chondroclasts. Histomorphometry revealed a significantly decreased trabecular bone volume (BV), but, surprisingly, also a marked increase in the number of osteoblasts, the rate of bone turnover, and the amount of mineralizing surface (MS). However, monitoring of bone density in the proximal tibias of the TG mice with peripheral quantitative computed tomography (pQCT) failed to reveal statistically significant changes in bone density. Similarly, no statistically significant alterations were observed in biomechanical testing at the age of 7 months. The increases in parameters of bone formation triggered by increased cathepsin K expression is an example of the tight coupling of bone resorption and formation during the bone-remodeling cycle. PMID- 11499868 TI - Reconstituting telomerase activity using the telomerase catalytic subunit prevents the telomere shorting and replicative senescence in human osteoblasts. AB - The rate of bone formation is largely determined by the number of osteoblasts, which in turn is determined by the rate of replication of progenitors and the life span of mature cells, reflecting the timing of death by apoptosis. However, the exact age-dependent changes of the cellular activity, replicative potential, and life span of osteoblasts have not been investigated to date. Here, we present evidence that the cellular activity, telomere lengths, and replicative life span of osteoblastic cells obtained from juxta-articular bone marrow gradually decrease with the advance of donor age. Recently, telomerase reverse transcriptase (hTERT) has been identified as a human telomerase catalytic subunit. We transfected the gene encoding hTERT into telomerase-negative human osteoblastic cells from donors and osteoblastic cell strain NHOst 54881 cells and showed that expression of hTERT induces telomerase activity in these osteoblastic cells. In contrast to telomerase-negative control cells, which exhibited telomere shortening and senescence after 10-15 population doublings, telomerase-expressing osteoblastic cells had elongated telomere lengths and showed continued alkaline phosphatase activity and procollagen I C-terminal propeptide (PICP) secretion for more than 30 population doublings. These results indicate that osteoblasts with forced expression of hTERT may be used in cell-based therapies such as ex vivo gene therapy, tissue engineering, and transplantation of osteoblasts to correct bone loss or osteopenia in age-related osteoporotic diseases. PMID- 11499869 TI - Dual mechanism of intercellular communication in HOBIT osteoblastic cells: a role for gap-junctional hemichannels. AB - Intercellular communication allows tissue coordination of cell metabolism and sensitivity to extracellular stimuli. Paracrine stimulation and cell-to-cell coupling through gap junctions induce the formation of complex cellular networks, which favors the intercellular exchange of nutrients and second messengers. Intercellular Ca2+ signaling was investigated in human osteoblast-like initial transfectant (HOBIT) cells, a human osteoblastic cell line in which cells retain most of the osteoblastic differentiation markers. HOBIT cells express connexin43 (Cx43) clustered at the cell-to-cell boundary and display functional intercellular coupling as assessed by the intercellular transfer of Lucifer yellow. Mechanical stimulation of a single cell induced a wave of increased Ca2+ that was radially propagated to surrounding cells. Treatment of cells with thapsigargin blocked mechanically induced signal propagation. Intercellular Ca2+ spreading and dye transfer were inhibited by 18alpha-glycyrrhetinic acid (18-GA), showing the involvement of gap junctions in signal propagation. Pretreatment of cells with suramin or with apyrase decreased the extent of wave propagation, suggesting that ATP-mediated paracrine stimulation contribute to cell-to-cell signaling. The functional expression of gap-junctional hemichannels was evidenced in experiments of Mn2+ quenching, extracellular dye uptake, and intracellular Ca2+ release, activated by uptake of inositol 1,4,5-trisphosphate (InsP3) from the external medium. Gap-junctional hemichannels were activated by low extracellular Ca2+ concentrations and inhibited by 18-GA. A role for Cx hemichannels in adenosine triphosphate (ATP) release and paracrine stimulation is suggested. PMID- 11499870 TI - Characterization of tissue transglutaminase in human osteoblast-like cells. AB - Tissue transglutaminase (tTG) is a calcium-dependent and guanosine 5' triphosphate (GTP) binding enzyme, which catalyzes the post-translational modification of proteins by forming intermolecular epsilon(gamma-glutamyl)lysine cross-links. In this study, human osteoblasts (HOBs) isolated from femoral head trabecular bone and two osteosarcoma cell lines (HOS and MG-63) were studied for their expression and localization of tTG. Quantitative evaluation of transglutaminase (TG) activity determined using the [1,4 14C]-putrescine incorporation assay showed that the enzyme was active in all cell types. However, there was a significantly higher activity in the cell homogenates of MG-63 cells as compared with HOB and HOS cells (p < 0.001). There was no significant difference between the activity of the enzyme in HOB and HOS cells. All three cell types also have a small amount of active TG on their surface as determined by the incorporation of biotinylated cadaverine into fibronectin. Cell surface related tTG was further shown by preincubation of cells with tTG antibody, which led to inhibition of cell attachment. Western blot analysis clearly indicated that the active TG was tTG and immunocytochemistry showed it be situated in the cytosol of the cells. In situ extracellular enzyme activity also was shown by the cell-mediated incorporation of fluorescein cadaverine into extracellular matrix (ECM) proteins. These results clearly showed that MG-63 cells have high extracellular activity, which colocalized with the ECM protein fibronectin and could be inhibited by the competitive primary amine substrate putrescine. The contribution of tTG to cell surface/matrix interactions and to the stabilization of the ECM of osteoblast cells therefore could by an important factor in the cascade of events leading to bone differentiation and mineralization. PMID- 11499871 TI - A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in vitro. AB - Breast cancer has a predilection for spreading to bone. The mechanism of preferential metastasis of breast cancer to bone is unknown. We hypothesize that breast cancer cells that develop bone metastases have the capacity to facilitate their colonization in bone. To examine this hypothesis, we established bone seeking (MDA-231BO) and brain-seeking (MDA-231BR) clones of the human breast cancer cell line MDA-MB-231 by repeated sequential passages in nude mice and in vitro of metastatic cells obtained from bone and brain metastases, respectively. These clones were examined for distinguishing biological characteristics and compared with the MDA-231 parental cells (MDA-231P) in vivo and in vitro. Both the MDA-231BR and the MDA-231BO showed identical tumorigenicity to MDA-231P at the orthotopic site. MDA-231P that was inoculated into the heart developed metastases in bone, brain, ovary, and adrenal glands. On the other hand, MDA 231BO exclusively metastasized to bone with larger osteolytic lesions than MDA 231P. MDA-231BR exclusively disseminated to brain and failed to develop bone metastases. In culture, MDA-231BO produced greater amounts of parathyroid hormone related protein (PTH-rP) than MDA-231BR and MDA-231P in the absence or presence of transforming growth factor beta (TGF-beta). Furthermore, the anchorage independent growth of MDA- 231BO in soft agar was not inhibited by TGF-beta, whereas TGF-beta profoundly inhibited the growth of MDA-231P and MDA-231BR. Insulin-like growth factor I (IGF-I) markedly promoted the anchorage-independent growth of MDA-231BO, whereas marginal or no stimulation was observed in MDA-231BR or MDA-231P, respectively. Our data suggest that these phenotypic changes allow breast cancer cells to promote osteoclastic bone resorption, survive, and proliferate in bone, which consequently leads to the establishment of bone metastases. PMID- 11499872 TI - Colocalization of glucocorticoid and mineralocorticoid receptors in human bone. AB - Osteoporosis is a poorly understood but common complication of glucocorticoid therapy. The actions of glucocorticoids are mediated via glucocorticoid receptors (GRs), but in vitro, glucocorticoids also can bind to mineralocorticoid receptors (MRs). It is not known if MR protein is present in human bone and little is known of GR isoform expression (GRalpha and GRbeta). GR and MR protein expression and possible sites of action were investigated in neonatal rib and adult iliac crest biopsy specimens using antibodies specific for MR, GRalpha, and GRalphabeta. Colocalization [MR GRalpha] [MR GRalphabeta] was performed using fluorescent conjugated secondary antibodies. GRalpha, GRbeta, and MR show distinct but overlapping patterns of expression, suggesting important functions for each receptor type. Osteoclasts showed no staining for GRalpha but strong staining for GRalphabeta, indicating expression of GRbeta and a specific role in addition to antagonizing the transcriptional activity of GRalpha. MR also was observed in osteoclasts and colocalized with GRalphabeta. Coexpression of MR, GRalpha, and GRalphabeta was seen in osteoblasts. Reverse-transcription-polymerase chain reaction (RT-PCR) of cultured osteoblast RNA confirmed expression of both GRalpha and GRbeta. Osteocytes stained with MR, GRalpha, and GRalphabeta antibodies but to a lesser degree than osteoblasts. In the neonatal rib cartilage, staining for GRalpha, GRalphabeta, and MR was present in approximately one-half of the resting and hypertrophic chondrocytes and in most of proliferating chondrocytes and chondrocytes within the mineralizing matrix. Identification of MR raises the possibility that the physiological and pharmacologic effects of glucocorticoids on bone may be mediated via MR as well as GR and that GRalpha, GRbeta, and MR synergize to influence corticosteroid metabolism in human bone. PMID- 11499873 TI - Bone reinnervation after fracture: a study in the rat. AB - Reinnervation after tibial fracture in the rat was studied by analyzing the occurrence of growth-associated protein 43 (GAP-43), a marker for regenerating nerve fibers, and protein gene product 9.5 (PGP-9.5), a marker for mature nerve fibers, by immunohistochemistry. At 3 days postfracture, GAP-43--immunoreactive nerve fibers were first observed in the fracture hematoma and periosteum. At 7 days postfracture, abundant sprouting of GAP-43--positive fibers was seen in the callus, hyperplastic periosteum, and edge of fibrocartilage. In the latter region, the nerve fibers were nonvascular, showing dense ramifications and terminal sprouting close to chondroid cells. At 14 days and 21 days postfracture, many GAP-43--positive fibers were still sprouting into the fibrocartilage and new woven bone. Fine varicose GAP-43--positive fibers also were present in the bone marrow. In contrast to GAP-43, PGP-9.5-positive nerve fibers were observed only occasionally at 3 days postfracture but gradually increased in number from day 14 to 21. Our study shows that intense nerve regeneration occurs in early fracture healing partly unrelated to neovascularization. Considering that neuronal mediators have been shown to participate in local bone formation and resorption, the nerve regeneration observed may prove to be essential for delivery of neuronal mediators required for normal callus formation and/or neovascularization. PMID- 11499874 TI - Trabecular structure assessment in lumbar vertebrae specimens using quantitative magnetic resonance imaging and relationship with mechanical competence. AB - The purpose of this study was to use quantitative magnetic resonance imaging (MRI; high-resolution [HR] and relaxometry) to assess trabecular bone structure in lumbar vertebrae specimens and to compare these techniques with bone mineral density (BMD) in predicting stress values obtained from mechanical tests. Fourteen vertebral midsagittal sections from lumbar vertebrae L3 were obtained from cadavers (aged 22-76 years). HR images with a slice thickness of 300 microm and an in-plane spatial resolution of 117 microm2 x 117 microm2 were obtained. Transverse relaxation time T2' distribution was measured by using an asymmetric spin-echo (ASE) sequence. Traditional morphometric measures of bone structure such as apparent trabecular bone fraction (app. BV/TV), apparent trabecular bone number (app. Tb.N), apparent trabecular bone separation (app. Tb.Sp), and apparent trabecular bone thickness (app. Tb.Th) as well as the directional mean intercept length (MIL) were calculated. Additionally, BMD measurements of these sections were obtained by dual-energy X-ray absorptiometry (DXA) and biomechanical properties such as directional stress values (to fracture) were determined on adjacent specimens. With the exception of T2', all morphological parameters correlated very well with age, BMD, and stress values (R between 0.79 and 0.92). However, in the direction perpendicular to the magnetic field, T2' values enhanced the adjusted R2 correlation value with horizontal (M/L) stress values in addition to BMD from 0.70 to 0.91 (p < 0.05). PMID- 11499875 TI - Digital topological analysis of in vivo magnetic resonance microimages of trabecular bone reveals structural implications of osteoporosis. AB - Osteoporosis is a disease characterized by bone volume loss and architectural deterioration. The majority of work aimed at evaluating the structural implications of the disease has been performed based on stereologic analysis of histomorphometric sections. Only recently noninvasive imaging methods have emerged that provide sufficient resolution to resolve individual trabeculae. In this article, we apply digital topological analysis (DTA) to magnetic resonance microimages (mu-MRI) of the radius obtained at 137 x 137 x 350 microm3 voxel size in a cohort of 79 women of widely varying bone mineral density (BMD) and vertebral deformity status. DTA is a new method that allows unambiguous determination of the three-dimensional (3D) topology of each voxel in a trabecular bone network. The analysis involves generation of a bone volume fraction map, which is subjected to subvoxel processing to alleviate partial volume blurring, followed by thresholding and skeletonization. The skeletonized images contain only surfaces, profiles, curves, and their mutual junctions as the remnants of trabecular plates and rods after skeletonization. DTA parameters were compared with integral BMD in the lumbar spine and femur as well as MR-derived bone volume fraction (BV/TV). Vertebral deformities were determined based on sagittal MRIs of the spine with a semiautomatic method and the number of deformities counted after threshold setting. DTA structural indices were found the strongest discriminators of subjects with deformities from those without deformities. Subjects with deformities (n = 29) had lower topological surface (SURF) density (p < 0.0005) and surface-to-curve ratio (SCR; a measure of the ratio of platelike to rodlike trabeculae; p < 0.0005) than those without. Profile interior (PI) density, a measure of intact trabecular rods, was also lower in the deformity group (p < 0.0001). These data provide the first in vivo evidence for the structural implications inherent in postmenopausal osteoporosis accompanying bone loss, that is, the conversion of trabecular plates to rods and disruption of rods due to repeated osteoclastic resorption. PMID- 11499876 TI - Variability in skeletal mass, structure, and biomechanical properties among inbred strains of rats. AB - The aim of this study was to assess the usefulness of the inbred rat model for studies of genetic influences on skeletal fragility. We characterized bone mass, geometry, and skeletal biomechanics in 11 inbred strains of rats. This study showed that considerable variation exists in bone structure, areal bone mineral density (aBMD), and fragility phenotypes among inbred strains of rats. Interestingly, the variability in skeletal phenotypes in rats was site specific, suggesting that no single gene regulates skeletal fragility at all sites. For instance, the Copenhagen 2331 (COP) strain had the greatest biomechanical properties in the femoral neck but only modest bone strength at the femoral midshaft, compared with other strains. Consequently, COP rats appear to have alleles that specifically enhance femoral neck biomechanical properties and may serve as a model for studying genetic influences on hip strength. The Brown Norway (BN) and Fischer 344 (F344) strains may provide models for vertebral fragility because each has relatively fragile lumbar vertebrae. The F344 rats also had the most fragile femora and, thus, appear to carry alleles that cause overall skeletal fragility. We identified two inbred rat crosses that will facilitate the study of genetic influences on skeletal fragility at clinically relevant skeletal sites: Lewis (LEW) with F344 (primarily for vertebral fragility) and COP with DA (primarily for femoral neck fragility). The results strongly suggest that selected crosses of inbred strains of rats will provide useful models for studying genetic influences on bone strength and structure. PMID- 11499877 TI - Influence of the upper femur and pelvic geometry on the risk and type of hip fractures. AB - The geometry of the upper femur has been reported to associate with the hip fracture risk in postmenopausal women. However, these associations seem to be partly conflicting, probably because of differences in measurement setup. Here, we compared the upper femur and pelvic geometries of 70 hip fracture patients (46 cervical and 24 trochanteric fractures) and 40 age-adjusted controls based on plain anteroposterior radiographs, eliminating the possible sources of inaccuracy as far as possible by using a standardized patient position and calibrated dimension measurements by digital image analysis. The femoral neck/shaft angle (NSA) was larger in the fracture patients compared with the controls (p < 0.001). The fracture group had thinner cortices in the upper femur than the controls (p < 0.001). The femoral shaft diameter (FSD; p < 0.001), trochanter width (TW; p < 0.01), and the pelvic dimensions, that is, the smallest outer pelvic diameter (SOPD; p < 0.01) and the largest inner pelvic diameter (LIPD; p < 0.05) were smaller in the fracture group. Comparing the fracture types, we found NSA larger in the cervical hip fracture patients than in the patients with a trochanteric fracture (p < 0.01). The femoral neck/shaft cortex ratio was lower and the FSD was smaller in the cervical hip fracture group (p < 0.05). Acetabular width (AW) was greater and the SOPD was wider in the cervical fracture patients (p < 0.01). We concluded that the upper femur and pelvic dimensions as defined from calibrated and position-standardized plain radiographs are useful in the evaluation of hip fracture risk and fracture type. PMID- 11499878 TI - The development of metaphyseal cortex--implications for distal radius fractures during growth. AB - Fractures of the distal radial metaphysis are very common in otherwise healthy children. The reasons for this high fracture incidence are not entirely clear. To address this problem, we undertook a detailed analysis of distal radius development using peripheral quantitative computed tomography (pQCT) at a site 4% proximal to the radial articular surface. The study population comprised 337 healthy children and adolescents (aged 6-18 years; 171 girls) and 107 adults (aged 29-40 years; 88 women). Total volumetric bone mineral density (vBMD) remained stable at about 70% of the adult value between the ages of 6-7 years and 14-15 years in both genders. Cortical thickness increased little between 6-7 years and 12-13 years in girls and 14-15 years in boys. Strength-Strain Index (SSI; a parameter combining geometry and density) was still at only 20% of the adult value in girls aged 10-11 years and at 21% of the adult level in boys aged 12-13 years. At these ages, factors that contribute to the mechanical challenge to the distal radius in case of a fall (forearm length and body weight) had already reached 49% and 36% of the adult value in girls and boys, respectively. The shaping of the distal radius cortex (metaphyseal inwaisting) was assessed by analyzing the decrease in cross-sectional bone size between adjacent bone slices in a separate population of 44 children (aged 8-19 years; 26 girls). The rates of periosteal resorption and endocortical apposition were estimated to average 8 microm/day and 10 microm/day, respectively, during the growth period. In conclusion, during growth the increase in distal radius strength lags behind the increase in mechanical challenges caused by a fall, because metaphyseal cortical thickness does not increase sufficiently. The endocortical apposition rate is already very high at that site and apparently cannot be further increased to levels that would be necessary to keep bone strength adapted to the mechanical requirements. PMID- 11499879 TI - Selenium deficiency-induced growth retardation is associated with an impaired bone metabolism and osteopenia. AB - Although the importance of selenium for bone metabolism is unknown, some clinical conditions such as Kashin-Beck osteoarthropathy have been associated with selenium deficiency. Although selenium deficiency induces growth retardation in rats, it has not been established whether this growth inhibition is associated with changes in bone metabolism. We investigated the effect of selenium deficiency on bone metabolism in growing male rats fed a selenium-deficient diet for two generations (Se-). In Se- rats, erythrocyte glutathione peroxidase activity and plasma selenium concentration were strongly reduced compared with pair-fed selenium-adequate rats (Se+). Weight and tail length were reduced by 31% and 13% in the Se- rats, respectively (p < 0.001). The Se- diet was associated with a 68% reduction of pituitary growth hormone (GH; p = 0.01) and a 50% reduction of plasma insulin-like growth factor I (IGF-I; p < 0.001). Plasma calcium was lower and urinary calcium concentration was greater in Se- rats. This group had a 2-fold increase in parathyroid hormone (PTH) and 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] in plasma. Plasma osteocalcin and urinary deoxypyridoline were reduced by 25% and 57% in the Se- rats (p < 0.001). Selenium deficiency resulted in a 23% and 21% reduction in bone mineral density (BMD) of the femur and tibia (p < 0.001) and this effect persisted after adjustment for weight in a linear regression model. A 43% reduction in trabecular bone volume of the femoral metaphysis (p < 0.001) was found in Se- rats. This experimental study shows that growth retardation induced by selenium deficiency is associated with impaired bone metabolism and osteopenia in second-generation selenium-deficient rats. PMID- 11499880 TI - Pseudo-(tumor-induced) rickets. AB - An athletic 8-year-old boy developed severe muscle weakness over 2 years. At the age of 10 years, investigation for possible neuromuscular disease disclosed hypophosphatemia (1.8 mg/dl) and rickets. There was selective renal tubular wasting of inorganic phosphate (Pi) but no history of toxin exposure, familial bone or kidney disease, or biochemical evidence of vitamin D deficiency. Urine amino acid quantitation was unremarkable. Serum 1,25-dihydroxyvitamin D [1,25(OH)2D] concentration was in the lower half of the reference range. Our presumptive diagnosis was tumor-induced rickets; however, physical examination and bone scanning in search of a neoplasm were unrevealing. Soon after 1,25(OH)2D3 and Pi treatment began, muscle strength improved considerably. After 6 months of therapy, radiographic abnormalities were substantially better. During the next 6 years, physical examinations, a second bone scan, whole-body and nasal sinus magnetic resonance imaging, and octreotide scintigraphy were unremarkable. When his physes fused at the age of 16 years, assessment of his course showed excellent control of his rickets requiring decreasing doses of medication. Furthermore, fasting serum Pi levels and tubular maximum phosphorus/glomerular filtration (TmP/ GFR) values had increased steadily and normalized after 3 years of treatment. Accordingly, therapy was stopped. Seven months after stopping medication, he continues to feel completely well. Fasting serum Pi levels, TmP/GFR, other biochemical parameters of bone and mineral homeostasis, creatinine clearance, and renal sonography are normal. Neither spontaneous or pharmacologic cure of tumor-induced rickets or osteomalacia nor a patient matching ours has been reported. His disorder, which we call pseudo-(tumor-induced) rickets, should be considered when investigation for oncogenic rickets or osteomalacia discloses no causal lesion. Consequently, prolonged medical therapy and futile searches for a neoplasm may be avoided. PMID- 11499881 TI - Detection of marijuana use by oral fluid and urine analysis following single-dose administration of smoked and oral marijuana. AB - We compared oral fluid testing to urine testing in subjects who were administered single doses of marijuana by smoked and oral routes. Oral fluid specimens were collected with the Intercept DOA Oral Specimen Collection Device, screened for THC with the Cannabinoids Intercept MICRO-PLATE Enzyme Immunoassay (EIA) utilizing a 1.0-ng/mL cutoff concentration, and confirmed for THC by gas chromatography-tandem mass spectrometry (GC-MS-MS) with a 0.5-ng/mL cutoff concentration. Urine specimens were screened for 11-nor-carboxy-delta9 tetrahydrocannabinol (THCCOOH) by immunoassay utilizing a 50-ng/mL cutoff concentration and confirmed for THCCOOH by GC-MS with a 15-ng/mL cutoff concentration. Oral fluid specimens tested positive following smoked marijuana (N = 10) consecutively for average periods (+/-SEM; range) of 15 (+/-2; 1-24) and 13 h (+/-3; 1-24) by EIA and GC-MS-MS, respectively. The average THC detection times of the last oral fluid positive specimen following smoked marijuana by EIA and GC MS-MS were 31 (+/-9; 1-72) and 34 h (+/-11; 1-72), respectively. In comparison to oral fluid, urine specimens generally tested negative for THCCOOH immediately after marijuana use. The average times to detection of the first urine specimen positive for THCCOOH by EIA and GC-MS were 6 (+/-2; 1-16) and 4 h (+/-1; 2-8), respectively. Urine specimens tested positive consecutively for average periods of 26 (+/-9; 2-72) and 33 h (+/-10; 4-72) for EIA and GC-MS, respectively. The average THCCOOH detection times of the last specimen by EIA and GC-MS were 42 (+/ 10; 2-72) and 58 h (+/-6; 16-72), respectively. Considering the noninvasive nature of oral fluid collection and improved detection of recent marijuana use compared to urine testing, it was concluded that oral fluid testing for THC offers specific advantages over other means of marijuana testing when used in safety-sensitive testing programs. PMID- 11499882 TI - Analysis of pyrolysis products of dimethylamphetamine. AB - This study examines the pyrolysis products of dimethylamphetamine (DMAMP) and its pyrolysis mechanism. A sealed glass tube, in which DMAMP-HCI was placed, was wrapped with pyrolysis-foil and heated at the Curie point of the pyrolysis-foil. The pyrolysis products of DMAMP were detected by gas chromatography-mass spectrometry (GC-MS), headspace (HS)-GC-MS and liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). DMAMP-d6-HCl, in which all the hydrogen atoms of the two methyl groups of DMAMP were substituted with deuterium atoms, was pyrolyzed to investigate the elimination and transformation reactions of the methyl group. Methamphetamine (MAMP) and amphetamine (AMP) were produced via demethylation reaction by heating DMAMP, and the maximum amounts of MAMP and AMP were about 31.8% and 13.7% of the starting DMAMP at 358 degrees C and 386 degrees C, respectively. The reactions of a chlorine anion and DMAMP with a methyl group eliminated from the dimethylamino group of DMAMP resulted in the formation of methyl chloride and benzylethyltrimethylammonium, respectively. These results indicate that one of the demethylation and methylation reactions occurs in the form of a methyl cation. PMID- 11499883 TI - Laboratory analysis of remotely collected oral fluid specimens for opiates by immunoassay. AB - The performance characteristics of a method for detecting opiates (morphine, codeine, heroin, and 6-acetylmorphine [6-AM]) in oral fluid specimens were examined and compared with methods for urine specimens. The oral fluid was easily obtained using a simple device that collects between 1 and 1.5 mL of fluid for laboratory analysis. Simultaneously collected specimens from 60 known opiate abusers from a drug-treatment center were first tested using an immunoassay cutoff of 10 ng/mL in oral fluids and 2,000 ng/mL in urine. Using a second aliquot, opiate confirmation in urine was performed by gas chromatography-mass spectrometry (GC-MS) and in oral fluids by GC-MS-MS. The combined immunoassay and GC-MS-MS procedures were completed with less than 250 pL of oral fluid. Opiates identified in oral fluid specimens from heroin users included morphine, codeine, heroin, and 6-AM. The immunoassay was tested for precision, stability, and the effects of potential cross-reactants. The results yielded 93.6% agreement between oral fluid and urine, suggesting that oral fluid may be a reliable matrix for opiate detection. PMID- 11499884 TI - Investigation into some aspects of EMIT d.a.u., TLC, and GC-MS urinalysis of bromazepam. AB - Among the different 1,4-benzodiazepine urinary metabolites, those of bromazepam possess distinctive chemical features that may be used in their selective isolation and detection. The detection of bromazepam metabolites in urine was carried out using EMIT d.a.u., thin-layer chromatography (TLC), and gas chromatography-mass spectrometry (GC-MS). The positive EMIT d.a.u. benzodiazepine assay for bromazepam was found to be due to the 3-hydroxybromazepam (3HOB) metabolite. The detection by TLC and GC-MS was carried out after enzyme or acid hydrolysis of the glucuronide conjugates. Both the 2-amino-3-hydroxy5 bromobenzoylpyridine (AHBBP) metabolite and the acid hydrolysis product of 3-HOB, 2-amino-5-bromobenzoylpyridine (ABBP), were selectively detected by TLC. The bromazepam metabolites in urine could be both isolated and detected selectively by GC-MS in the presence of the metabolites of other 1,4-benzodiazepines that were sometimes used in combination with bromazepam. Both 3-HOB and AHBBP were detected by GC-MS only after trimethylsilyl (TMS) derivatization and not as the free compounds or the acetyl derivatives. Only ABBP was detected in three forms: ABBP, the TMS derivative, and the acetyl derivative. Evidence has been obtained from the enzyme hydrolysis and the TLC studies for the formation of the glucuronide conjugate of AHBBP at the 3-OH group rather than at the 2-NH2 group. All the results have been validated using reference 3-HOB and AHBBP. PMID- 11499885 TI - Comparison of HPLC and GC-MS methods for determination of embutramide (a component of Tanax or T-61) in biological specimens. AB - Tanax or T-61, a euthanasia solution commonly used in veterinary medicine, has been often involved in suicide attempts (humans) and malicious intoxications (animals). For forensic reasons, the identification of one or more of the three components (embutramide, mebenzonium iodide, and tetracaine hydrochloride) of Tanax is needed to confirm the hypothesis of intoxication. This study was performed with new high-performance liquid chromatographic and gas chromatographic-mass spectrometric methods to identify embutramide in biological matrices (blood, liver, kidney) from different animal species. The good sensitivity and specificity of both methods recommend their use in toxicological analysis in both human and veterinary medicine. PMID- 11499886 TI - Improved procedure for the analysis of gamma-hydroxybutyrate and ethylene glycol in whole blood. AB - The modification of a procedure originally developed for the analysis of ethylene glycol (EG) in serum was also found to permit the simultaneous analysis of gamma hydroxybutyrate (GHB) in whole blood. The primary feature of the EG procedure was that it employed a water scavenger, 2,2-dimethoxypropane, which reacted with water to produce volatile methanol. Water scavenging is a technique that could be adapted for the analysis of drugs such as GHB as their respective di-t butyldimethylsilyl derivatives. A close structural analogue of GHB, 2-hydroxy-3 methylbutyric acid, was successfully employed as the internal standard for both EG and GHB. The advantages of the modified procedure are that it is very quick and easy to perform and produces remarkably clean extracts for GHB, especially when compared to other liquid-liquid techniques. We have successfully applied this technique for the analysis of GHB and EG in several postmortem and driving under-the-influence cases. There is an apparently wide variability between levels of GHB that can be associated with impairment versus those levels that can be associated with death. PMID- 11499887 TI - The antispasmodic drug mebeverine leads to positive amphetamine results by fluorescence polarization immunoassay (FPIA)--studies on the toxicological analysis of urine by FPIA and GC-MS. AB - Mebeverine (Duspatal, MB), an antispasmodic drug, is the veratric acid ester of 4 [ethyl-[2-(4-methoxyphenyl)-1-methylethyl]amino]butan-1-ol (MB-OH), which is a N substituted ethylamphetamine derivative. MB is metabolized via ester hydrolysis to MB alcohol (MB-OH) and veratric acid. N-Dehydroxybutylation leads to methoxyethylamphetamine (MO-EA) and, after O-demethylation, to hydroxy EA (HO EA). N-Bisdealkylation leads to p-methoxyamphetamine (PMA). MO-EA and PMA are also known as designer drugs. Fluorescence polarization immunoassay (FPIA) and gas chromatographic-mass spectrometric studies on the toxicological analysis of MB after ingestion of a single 405-mg oral dose of MB were performed. We could show that intake of MB leads to positive FPIA results for amphetamine. The N dehydroxybutyl metabolites of MB, MO-EA, HO-EA, and the bis-dealkyl metabolite PMA should be responsible for the positive immunoassay results. Using our systematic toxicological analysis procedure, every positive amphetamine immunoassay could be explained by detection of MO-EA, HO-EA, and/or PMA. Misinterpretation of the origin of MO-EA, HO-EA, or PMA can be avoided by detecting the specific (non-dehydroxybutylated) metabolites of MB, which are excreted for a much longer time after ingestion. PMID- 11499888 TI - Importance of vacutainer selection in forensic toxicological analysis of drugs of abuse. AB - The enzymatic degradation of cocaine in blood samples, even during transport to a forensic laboratory, is a common problem in toxicological analysis. This can be avoided by the use of blood-sampling devices such as gray-top Vacutainers containing the cholinesterase inhibitor sodium fluoride. In the present study, which included 147 authentic cases, blood samples were collected into two different tubes, one containing fluoride/oxalate and one without stabilizing agents. In all cases, both samples were analyzed for drugs of abuse using Abbott FPIA immunoassays after precipitation and gas chromatography-mass spectrometry (GC-MS) for quantitative analysis. The cannabinoid immunoassay showed markedly lower values in the fluoride-containing samples; this was investigated further and could be explained by hemolysis of these samples. In addition, the concentrations of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid (THCCOOH) were lower in these samples. A stability study with the THCCOOH acyl glucuronide showed that it is unstable in unpreserved serum, which could explain our observation. GC-MS quantitative data for amphetamine and derivatives, opiates, delta9-tetrahydrocannabinol, and 11-hydroxy-delta9-tetrahydrocannabinol were essentially identical; however, they also differed substantially for cocaine, cocaethylene, ecgonine methylester, and benzoylecgonine. Unexpectedly, the concentrations of benzoylecgonine in unpreserved serum were almost half as high as in the fluoride-containing samples. PMID- 11499889 TI - Dermal exposure to strychnine. AB - A non-fatal case of strychnine poisoning through dermal exposure is described. About 24 h after cleaning up a strychnine spill, a 50-year-old woman presented to the emergency department with classical signs of strychnine poisoning, consisting of marked pain in the muscles of her lower limbs, dermal sensitivity, and stiffness in her jaw. Her treatment was intravenous fluid replacement and alkalinization in anticipation of potential renal failure due to rhabdomyolysis. Her plasma creatine kinase was elevated to 677 U/L with no rise in the heart specific MB fraction. Serum myoglobin level obtained retrospectively was 195 microg/L. Biological samples were taken approximately 28 h after her exposure. Strychnine was measured in plasma (196 ng/mL) and urine (6,850 ng/mL) by gas chromatography-mass spectrometry. The small amount of pheniramine in the plasma (35 ng/mL) and urine (1,255 ng/mL) is considered an inconsequential finding. PMID- 11499890 TI - Concentrations of cis(Z)-clopenthixol and trans(E)-clopenthixol in a lethal case involving zuclopenthixol, diazepam, and cyamemazine. AB - cis(Z)-Clopenthixol and trans(E)-clopenthixol were determined by gas chromatography-mass spectrometry and high-performance liquid chromatography-diode array detection in necropic samples from a postmortem case. The peripheral blood concentrations of cis(Z)-clopenthixol and trans(E)-clopenthixol were 278 and 177 ng/mL, respectively. The level of the active cis(Z)-isomer is within the toxic range. Other associated drugs' concentrations were within their therapeutic ranges. Postmortem redistribution of the drug and instability of the drug due to trans-isomerization were discussed. PMID- 11499891 TI - Heroin use by motorists in Sweden confirmed by analysis of 6-acetylmorphine in urine. PMID- 11499892 TI - Response to the presence of gamma-hydroxybutyric acid (GHB) in postmortem biological fluids. PMID- 11499893 TI - Informatics and scientific information exchange in forensic toxicology. PMID- 11499894 TI - Ethics for toxicologists: an examination of conscience. PMID- 11499895 TI - Alternate strategies for postmortem drug testing. PMID- 11499896 TI - Alternative specimens for workplace drug testing. AB - Recent advances in analytical techniques have enabled the detection of drugs and drug metabolites in alternative biological specimens for the purposes of workplace testing. A wide variety of specimens are available, each providing valuable information concerning prior or current drug use. The present focus is on oral fluid (saliva), hair, and sweat. An extensive evaluation by the Division of Workplace Programs of the Department of Health and Human Services is underway to determine the utility of these specimens in federally regulated programs. In future years, the testing of alternative specimens will expand our ability to understand the patterns of drug use and will become routine in all areas of forensic toxicology. PMID- 11499897 TI - Amphetamines: an update on forensic issues. AB - Methamphetamine is currently enjoying a resurgence of popularity as a recreational drug. It presents a number of challenges to the forensic toxicologist both analytically and interpretively, and these latter interpretive issues are considered here. This review also discusses the current popular syntheses which account for the widespread domestic synthesis of the drug; the demographics of methamphetamine use in the United States as assessed from the Drug Abuse Warning Network (DAWN) data; developments in research of the neurotransporter pharmacology of the drug and its implications for interpretive forensic toxicology; the psychomotor effects of the drug and its potential for cognitive and functional impairment; interpretive issues related to postmortem blood drug concentrations and how these are impacted by evidence for incomplete distribution and the potential for postmortem redistribution; and, finally, concerns caused by designer methamphetamine analogues. All data indicate that methamphetamine and its analogues will present significant interpretive challenges to forensic toxicologists as the popularity of the drug continues to grow. PMID- 11499898 TI - Genetic mechanisms for variability in drug response and toxicity. AB - It is now well established that many proteins involved in the metabolism or pharmacodynamic action of drugs and foreign compounds exhibit structural polymorphism and variation in their level of expression. This variation leads to dramatic phenotypic differences in response to medicines or susceptibility to carcinogenesis. Some of the changes in the phenotypic expression of proteins are secondary to variation in the nucleic acid sequence of their respective genes. The science of pharmacogenetics links differences in gene structure (polymorphism) with pharmacologic differences in drug action and disposition of foreign compounds. Through discussion of four examples, we will emphasize the variety of genetic mechanisms that can potentially influence the phenotypic response to xenobiotic challenge and pharmacotherapy. The first example illustrates how structural variation in the coding region of drug metabolizing enzymes influences risk of drug toxicity. A second example demonstrates how genetic variation can influence gene transcriptional regulation and how the resulting dysregulation may be linked to increased susceptibility to exposure linked cancer. The third example illustrates how genetic polymorphism can selectively influence the pharmacodynamic response to medication, and the final example of warfarin response illustrates how genetic variation in more than one gene can account for broad extremes in phenotypic response. PMID- 11499899 TI - Structural MRI as a tool for the study of neurotoxicity and neurodegenerative disorders. AB - High-resolution magnetic resonance (MR) imaging affords an unprecedented opportunity to study the severity and distribution of neurodegenerative changes in the human brain. By selecting specific MR sequence parameters (i.e., TE and TR), different MR signals can be received from different tissue types, such as gray and white matter. Through optimization of the contrast between different tissue types, the surfaces and internal structures of brain structures of special interest can be visualized and quantitated. Metrics such as two-dimensional areas, three-dimensional volumes, and three-dimensional shape characteristics have proven to be highly useful for quantitating the effects of toxins on the human brain. Among toxins, the effects of alcohol on the human brain have been most intensively studied using structural MR imaging. Volume losses in the cerebral cortex and other brain regions of interest have been carefully quantitated. However, because exposure to alcohol is almost always repeated over many years, the effects of normal aging must be carefully considered when making comparisons between diseased and healthy populations. In contrast to the literature on alcohol, structural MR imaging has been relatively underutilized in the study of drugs and other chemicals such as MPTP and other drugs of abuse that are toxic to special populations of neurons. However, as the resolution of structural MR continues to improve, the structural characteristics of such neuron populations will be visualized and quantitated, and successful use of structural MR imaging for the study of such toxins will become possible. PMID- 11499900 TI - fMRI: a new tool for the in vivo localization of drug actions in the brain. AB - Functional magnetic resonance imaging (fMRI), a still-emerging, non-invasive neuroimaging tool, has been applied to a wide range of questions in sensory, motor control, and cognitive psychology. Only more recently has it been applied to understand the sites and mechanisms of action of pharmacological agents within the human CNS. However, in so doing, a new set of problems and concerns surrounding the technique must be addressed because of the unique transduction mechanisms (both physiological and biophysical) that exist to produce the fMRI signal from the underlying neuronal activity. Experimental design and control issues become paramount in performing fMRI pharmacological protocols and in signal interpretation. With these caveats, the use of pharmacological agents with fMRI is likely to greatly increase in the near term as new questions about both brain physiology and neuropharmacological mechanisms become addressable for the first time. Examples are given using nicotine and cocaine as a prototypical agents. PMID- 11499901 TI - Intrahepatic biliary lesions after orthotopic liver transplantation. AB - Intrahepatic biliary lesions (IBL) are rare (2-9%) after orthotopic liver transplantation (OLT). The aim was to evaluate the incidence, etiology and outcome. In nine years, a total 532 OLTs were performed in 481 patients. Twenty four patients developed IBL. Eight were due to HAT, seven to ABOI, three to CDR and six to PI. The time until diagnosis of HAT is longest in patients (14+/-6) with IBL. ABOI is another cause of IBL. CDR is a rare cause of IBL, however when it takes place, patients must undergo Rtx. Finally, PI is a relevant cause of IBL. In order to suppress the incidence of IBL we should consider 1) the systematic use of Doppler-Ultrasound; 2) emergency reoperation of patients with HAT, 3) avoid ABOI in OLT; 4) Rtx in cases of CDR, and 5) OLT should still be performed as an emergency procedure. PMID- 11499902 TI - Sirolimus improves the two-year outcome of renal allografts in African-American patients. AB - The present study evaluated whether the addition of sirolimus to a cyclosporine (CyA)/prednisone (Pred) regimen mitigated the greater proclivity to acute rejection episodes and graft loss characteristic of African-American renal transplant recipients. Using Kaplan-Meier and log-rank tests, African-American renal transplant recipients treated with either CyA/Pred (n = 90) or sirolimus/CyA/Pred (n = 47) were compared with 120 Caucasian patients treated with sirolimus/CyA/Pred for 2-year rates of patient and graft survival as well as acute rejection episodes. Mean laboratory values were compared using analysis of variance and F-tests. Addition of sirolimus to the CyA/Pred regimen reduced the incidence of acute rejection episodes in African-Americans from 43.3% to 19.2% (P = 0.004), a value similar to Caucasian patients. The 97.9% 2-year graft survival rate among 47 African-American patients treated with sirolimus/CyA/Pred was significantly higher than the 85.6% rate shown among the 90 CyA/Pred-treated African-American transplant recipients (P = 0.0479) and similar to that in Caucasians. The 95.7% patient survival rate among the African-American sirolimus/CyA/Pred group was similar to the 97.8% rate in the African-American CyA/Pred cohort. Interestingly, there was no evident toxicity from the addition of sirolimus. The addition of sirolimus to a CyA-based regimen reduced acute rejection episodes and graft loss experienced by African-American renal transplant recipients. PMID- 11499903 TI - Chronic rejection in H-2 matched cardiac allografts: early emergence of vasculopathy, alloantibody, and accumulation of IFN-gamma and IL-10 mRNA. AB - Cardiac allograft vasculopathy (CAV) is one of the crucial problems of clinical heart transplantation. We have developed a novel model of murine cardiac allograft rejection, in which chronic rejection associated with CAV occurs in its natural course. In this study we analyzed the pathogenesis of chronic cardiac allograft rejection using an H-2 matched multiple minor histocompatibility antigen-mismatched combination, AKR (H-2k) to C3H (H-2k) recipient mice. All the cardiac allografts survived for more than 100 days but were rejected within 260 days post-transplant (n = 13; mean survival times +/- standard deviation = 189.0+/-72.0; median = 210). The heartbeats of the graft became gradually weaker throughout the duration of the rejection process. Serial histological analyses with hematoxylin and eosin, elastica van Gieson or Masson trichrome staining revealed mononuclear cell infiltration and intimal thickening (i.e. CAV) which started in most grafts at 2 weeks post-transplant. These pathological changes eventually developed to severe graft fibrosis, and the severity of these changes correlated with the deterioration of the heartbeats. Production of anti-donor antibodies in most recipients was detectable by 2 weeks post-transplant, it peaked before day 100, and subsided before rejection was complete in most grafts. Intragraft expression of IFN-gamma and IL-10 mRNA was demonstrated by reverse transcriptase-polymerase chain reaction during early periods post-transplant. In this study, we demonstrate a novel model feasible for analysis of chronic cardiac allograft rejection, in which the vascular rejection processes, including fibrosis and alloantibody production, can be tested from an early stage on, after transplantation. PMID- 11499904 TI - Identification of two down-regulated genes in rat liver allografts by mRNA differential display. AB - Total RNA differential display (DD) using random primers was performed for rat orthotopic liver transplantation (OLT) models. DA (RT1a) donor livers were transplanted into DA, PVG (RT1c), and LEW (RT1l) recipients: (1) syngeneic OLT (DA-DA): no rejection occurs; (2) allogeneic OLT (DA-PVG): rejection occurs, but is naturally overcome without immunosuppression; (3) allogeneic OLT (DA-LEW): animals die of acute rejection within 14 days. cDNA was isolated from selected bands, re-amplified for sequencing, and confirmed by Northern blots. Two down regulated genes were observed in day-7 allogeneic OLT livers (DA-PVG, DA-LEW), while they were consistently expressed in day-7 syngeneic OLT (DA-DA) livers. These two genes were identified as alpha-glutathione sulfotransferase (alpha-GST) Ya gene and estrogen sulfotransferase (EST), respectively. Northern blots confirmed that their expression was down-regulated in OLT (DA-PVG) livers on days 7-26 and gradually restored. The mRNA expression of GST and EST may be good markers to predict rejection or induction of tolerance. PMID- 11499905 TI - Bone marrow augmentation in kidney transplantation: a large animal study. AB - Specific immunomodulatory strategies are required to eliminate the need for lifelong dependence on debilitating immunosuppressants. One proposed strategy is to simultaneously transplant the kidney and infuse donor-specific bone marrow cells. We prospectively studied the effect of unmodified donor-specific bone marrow infusion (DSBMI) on rejection, infection, graft-versus-host disease (GvHD), and graft survival. We performed 57 kidney transplants in mixed lymphocyte culture (MLC)-reactive, outbred pigs. The groups of recipient pigs differed according to the use of (1) indefinite versus short-term tacrolimus based immunosuppression, (2) DSBMI, and (3) recipient preconditioning (RPC: whole body irradiation with 400 rads on day 0 and horse anti-pig thymocyte globulin (ATG) on days -2, -1, and 0). In all, we studied eight groups: group 1, nonimmunosuppressed control pigs (n = 8); group 2, nonimmunosuppressed DSBMI pigs (n = 7); group 3, nonimmunosuppressed RPC + DSBMI pigs (n = 5); group 4, tacrolimus (indefinite) pigs (n = 11); group 5, tacrolimus (10 days only) pigs (n = 5); group 6, DSBMI + tacrolimus (indefinite) pigs (n = 8); group 7, DSBMI + tacrolimus (10 days only) pigs (n = 6); and group 8, RPC + DSBMI + tacrolimus (indefinite) pigs (n = 7). DSBMI alone (group 2) or in combination with RPC (group 3) did not prolong graft survival, as compared with nonimmunosuppressed controls (group 1). In groups 1, 2, and 3, all but one pig died from rejection; in group 3 only, 45% of the pigs died from concurrent infection or GvHD, indicating that RPC in combination with DSBMI aggravated the risk of generalized infection and GvHD. Post-transplant immunosuppression--irrespective of indefinite or short-term administration--was required for prolonged graft survival. With indefinite use of immunosuppression, graft survival rates and death rates from rejection were not different for pigs with (group 6) versus without (group 4) DSBMI; however, the death rate from infection was higher in group 6, suggesting that the bone marrow inoculum increased the risk of systemic infection. With short-term use of immunosuppression, graft survival rates were higher and death rates from rejection lower for pigs with (group 7) versus without (group 5) DSBMI. But DSBMI and short-term immunosuppression (group 7) failed to prolong survival beyond that achieved with indefinite immunosuppression (groups 4 and 6). Although the combination of DSBMI and short-term immunosuppression (group 7) reduced the risk of infection, it did not avert severe rejection. The addition of RPC to DSBMI and indefinite immunosup- pression (group 8) significantly decreased graft survival, as compared with groups 4, 6, and 7. It also increased the incidence of death from rejection, GvHD, and infection, or a combination thereof. Unmodified DSBMI did not prolong graft survival after kidney transplantation, nor did it decrease the incidence of rejection. But it aggravated the risk of GvHD and infection. Short-term immunosuppression with DSBMI reduced the incidence of death from infection or GvHD, but it resulted in a higher incidence of death from rejection (as compared with indefinite use of immunosuppression). RPC, combined with DSBMI and indefinite immunosuppression, increased the death rate from rejection, GvHD, infection, or a combination thereof. In this large animal study, the effect of unmodified DSBMI has been disappointing. The search continues for the optimal way to successfully perform bone marrow augmentation in solid organ transplants. PMID- 11499906 TI - Do alcoholic liver transplantation candidates merit lower medical priority than non-alcoholic candidates? AB - Alcoholic patients are frequently regarded as responsible for their alcoholism and alcohol-related diseases, such as liver damage. These patients run the risk of receiving lower medical priority for liver transplantation than patients who are considered as not responsible for their liver damage. However, hardly any scientific research findings support this supposed responsibility of the alcoholic patient for his addiction and the related diseases. Many alcoholic patients have comorbid psychiatric disorders such as antisocial personality disorder, schizophrenia and social phobia, and these cormorbid diseases are often linked specifically and also in a neurobiological way to alcohol abuse. Furthermore, concepts such as responsibility and health have multiple dimensions, which can be contrasted against each other. Useful and fair criteria are presented for the assessment of responsibility for our health. PMID- 11499907 TI - The limits of philosophy in liver transplantation. AB - At the Editor's request, Dr. Beresford reviews the paper by Martens, with particular reference to the clinical literature on liver transplantation in alcoholism. He concludes that abstract concepts, such as responsibility for one's illness, may have some theoretical value in academic discussions but that no studies support any direct practical usefulness for them in patient selection for solid organ transplantation. Rather, a sizeable body of work delineates a practical clinical approach to the evaluation of alcohol-dependent persons who request liver transplantation. He highlights the relevant aspects of this work briefly. PMID- 11499908 TI - Pulmonary diffusion abnormalities in relation to cytomegalovirus antigenemia and cytomegalic endothelial cells in blood. AB - The pathophysiology of HCMV infection may involve many different organs including the lungs. In this study we investigated HCMV antigenemia levels and cytomegalic endothelial cells (CEC) in blood in relation to the pulmonary diffusion capacity. Patients with high HCMV antigenemia (> or = 100 pp65+ PMNs/50.000) (n = 8) showed a more extensive decrease in the membrane factor (Dm) than patients with lower levels of HCMV antigenemia (n = 7). The decline of the diffusion capacity of the alveolar capillary membrane (KCOc) and of the pulmonary capillary volume (Vcap) was the same in both groups. Four out of nine patients had CEC in the range of 0.22 CEC/ml to 30.26 CEC/ml. All the HCMV patients showed a decreased KCOc together with a decrease of Dm and Vcap but no difference was observed between patients with and without CEC. We conclude that a higher viral load is associated with a more extensive decrease in the membrane factor and therefore with more subclinical pneumonitis. No relation was observed between CEC and pulmonary dysfunction. Therefore, we postulate that CEC levels are related indirectly to subclinical pneumonitis mediated via the viral load. PMID- 11499909 TI - Relevance of two-stage total hepatectomy and liver transplantation in acute liver failure and severe liver trauma. AB - Emergency liver transplantation frequently is the only life-saving procedure in cases of acute liver failure. It remains unclear whether emergency hepatectomy with portocaval shunt followed by liver transplantation as a two-stage procedure should be performed in cases in which a donor organ is not yet available. It has been stated that "toxic liver syndrome" could be treated by means of this strategy. From 1990 to 1995 we performed emergency hepatectomies in eight cases of acute liver failure or traumatic liver rupture with exsanguinating bleeding. In six cases we were able to perform a subsequent liver transplantation. Five of the six patients who underwent an emergency hepatectomy died. Emergency hepatectomy led to a significant increase in epinephrine dosage until the transplantation was performed. Only after transplantation did the need for epinephrine therapy decrease. The need for oxygen support did not change during the entire observation period. Plasmatic coagulation was stabilized by substitution, showing significantly higher values at 24 h after transplantation than at 48 h before transplantation. Fibrinogen increased significantly after transplantation in this group of patients. The experiences gathered at our clinic, however, do not show advantages that would allow a recommendation of emergency hepatectomy and subsequent liver transplantation as a two-stage procedure except for situations of severe and uncontrollable hepatic bleeding. Considering the progressive destabilization of our patients, fast procurement of donor organs seems to be of imminent importance for the outcome. PMID- 11499910 TI - Selective cytotoxicity of MIA Pa Ca-2 conditioned medium to acinar cells: a novel approach to reduce acinar cell contaminants in isolated islet preparations from BALB/c mice. AB - Contamination of acinar cells in islet preparations has been shown to affect islet viability, functionality and yield adversely. Therefore, a strategy which would reduce acinar contamination in islet preparations is much sought. We here demonstrate selective cytotoxicity of conditioned medium (CM) of MIA Pa Ca-2 (human pancreatic carcinoma) cells to acinar cells and suitability of this approach as a simple method of obtaining a pure islet population without affecting their viability and yield. When isolated, islet preparations from BALB/c mice were exposed to conditioned media of MIA Pa Ca-2, acinar cells underwent extensive degeneration to yield 80-85% pure islet population, and islets showed comparable viability to controls not exposed to conditioned media. They also maintained their normal morphology, as assessed by digital image analysis. Islets treated with a conditioned medium also preserved in vitro insulin secretion. Flow cytometric analysis of acinar cells treated with conditioned media revealed accelerated DNA damage (45%), compared to controls (22%). Results emphasize the role of factors secreted by MIA Pa Ca-2 cells in inducing selective toxicity to acinar cells. PMID- 11499911 TI - Multiorgan donation from a donor with unrecognized ornithine transcarbamylase deficiency. AB - Ornithine transcarbamylase (OTC) deficiency, the most common inherited urea cycle disorder, shows a spectrum of severity ranging from severe neonatal hyperammonemic coma to no symptoms among adults. We report on the multiorgan procurement from a donor who died of cerebral edema due to unrecognized late onset OTC deficiency. The donor's OTC deficiency was diagnosed retrospectively since the liver graft recipient developed cerebral edema postoperatively due to hyperammonemia. Plasma ammonia was extremely elevated (3793 micromol/l), but was not accompanied by general liver dysfunction. Post mortem, the diagnosis of OTC deficiency was established by enzyme and molecular analysis in a biopsy of the transplanted liver. In contrast to the fatal course of the liver graft recipient, the kidney, lung, and heart transplantations were successful. Ten months after transplantation these recipients were alive and showed good graft function. This case demonstrates the importance of careful donor evaluation, particularly if the donor's cause of death is obscure. PMID- 11499912 TI - Nocardiosis revealed by thyroid abscess in a liver--kidney transplant recipient. AB - Nocardiosis is a life-threatening infection, particularly among immunocompromised patients, which usually affects lungs, skin and central nervous system. We report a case of disseminated nocardiosis revealed by suppurative thyroiditis in a liver kidney transplant recipient with poor nutritional status at the time of infection. Nocardia Asteroides was isolated from fine-needle aspiration material of the thyroid abscess. Clinical manifestations resolved after surgical drainage of the thyroid abscess, prolonged antibiotherapy and diminution of immunosuppressive regimen. Clinicians should be aware of this entity, as Nocardia Asteroides may need more than 5 days of culture to be isolated. PMID- 11499913 TI - Pump upgrade for machine perfusion at the Freeman Hospital in Newcastle. PMID- 11499914 TI - Fungal hydrophobins in medical and technical applications. AB - Class I and class II hydrophobins are small secreted fungal proteins that self assemble at hydrophilic-hydrophobic interfaces into amphipathic films. Apart from eight conserved cysteine residues, the amino acid sequences between and within both classes have diverged considerably, and this is reflected in the biophysical properties of these proteins. For instance, assemblages of class I hydrophobins are highly insoluble, while those of class II hydrophobins readily dissolve in a variety of solvents. The properties of hydrophobins make them interesting candidates for use in a wide range of medical and technical applications. Each application has its own requirements, which may be met by using specific natural variants of hydrophobins or by modifying hydrophobins chemically or genetically. Applications also require high production systems for hydrophobins. In this respect, filamentous fungi that naturally secrete hydrophobins into the medium seem to be the hosts of choice. PMID- 11499915 TI - Bioconversion of limonene to increased concentrations of perillic acid by Pseudomonas putida GS1 in a fed-batch reactor. AB - Pseudomonas putida GS1 is able to convert limonene to perillic acid (up to 64 mM,(11 g/l) when the bacteria is cultivated in fed-batch culture with non limiting amounts of glycerol. ammonium, and limonene. P. putida GS1 can use p cymene as a single source of carbon and energy, and the enzymes that are responsible for the conversion of limonene to perillic acid belong to the degradation pathway of p-cymene. The p-cymene pathway of P putida GS1 is very similar, if not identical, to the cym pathway of P. putida F1. The latter strain, and a recombinant Escherichia coli strain that carried the genes of the cym pathway of P. putida Fl, also converted limonene to perillic acid. However, the final concentrations that were obtained in batch cultures with these two strains were lower than those obtained with P. putida GS1. PMID- 11499916 TI - Effect of directional switching frequency on toluene degradation in a vapor-phase bioreactor. AB - A potential method to improve biomass distribution and the stability of vapor phase bioreactors is to operate them in a directionally switching mode such that the contaminant air stream direction is periodically reversed through the reactor. In this study, the effect of switching frequency (SF) on bioreactor performance and biodegradation activity was investigated at 1-, 3- and 7-day SFs using toluene as a model compound. Rapid losses of biodegradation capacity and serious bioreactor instability were observed in the bioreactor operated at a 1 day SF. It is hypothesized that the frequent dynamic loading conditions at the 1 day SF hindered biofilm development and ultimately bioreactor stability. In contrast, bioreactors operated at the 3- and 7-day SFs achieved overall removal efficiencies of greater than 99% for 72 and 59 days of operation, respectively. Following each air-stream reversal, the bioreactor operated at the 7-day SF required 48 h to fully restore biodegradation capacity in the inlet bioreactor section. The 1-day SF bioreactor required no such reacclimation period. The toluene-degrading activity in the inlet section of the 7-day SF bioreactor dropped by 71% during the 7-day cycle, whereas it decreased by only 11% in the inlet of the 3-day SF bioreactor. These declines suggest that continuous or near continuous exposure to toluene can inhibit microbial activity. Of the three SFs examined, the 3-day SF yielded the most efficient bioreactor performance by balancing reacclimation requirements with biodegradation activity losses. PMID- 11499917 TI - Production and characterization of anti-nisin Z monoclonal antibodies: suitability for distinguishing active from inactive forms through a competitive enzyme immunoassay. AB - As a pre-requisite to monoclonal antibody development, an efficient purification strategy was devised that yielded 72 mg of nisin Z from 14.5 1 of Lactococcus lactis subsp. lactis biovar. diacetylactis UL 719 (L. diacetylactis UL719) culture in supplemented whey permeate. Specific monoclonal antibodies (mAbs) were produced in mice against the purified nisin Z using keyhole limpet hemocyanin as a carrier protein. These antibodies did not recognize nisin A, suggesting that the asparagine residue at position 27 is involved in antibody recognition to nisin Z. However, the high reactivity of mAbs against biologically inactive nisin Z degradation products, produced during storage of freeze-dried pure nisin Z at 70 degrees C, indicated that the dehydroalanine residue at position 5 (Dha5), required for biological activity, is not necessary in nisin Z recognition by the mAb. A competitive enzyme immunoassay (cEIA) using the specific anti-nisin Z mAb was developed and used for rapid and sensitive detection and quantification of nisin Z in fresh culture supernatant, milk and whey. Detection limits of 78 ng/ml in phosphate-buffered saline, 87 ng/ml in culture supernatant, 106 ng/ml in milk and 90.5 ng/ml in whey were obtained for this assay. The cEIA using specific mAbs can be used to quantify nisin Z in food products. PMID- 11499918 TI - Use of catabolite repression mutants for fermentation of sugar mixtures to ethanol. AB - Use of agricultural biomass, other than corn-starch, to produce fuel ethanol requires a microorganism that can ferment the mixture of sugars derived from hemicellulose. Escherichia coli metabolizes a wide range of substrates and has been engineered to produce ethanol in high yield from sugar mixtures. E. coli metabolizes glucose in preference to other sugars and, as a result, utilization of the pentoses in hemicellulose-derived sugar mixtures is delayed and may be incomplete. Residual sugar lowers the ethanol yield and is problematic for downstream processing of fermentation products. Therefore, a catabolite repression mutant that simultaneously utilizes glucose and pentoses would be useful for fermentation of complex substrate mixtures. We constructed ethanologenic E. coli strains with a glucose phosphotransferase (ptsG) mutation and used the mutants to ferment glucose, arabinose, and xylose, singly and in mixtures, to ethanol. Yields were 87-94% of theoretical for both the wild type and mutants, but the mutants had an altered pattern of mixed sugar utilization. Phosphotransferase mutants metabolized the pentoses simultaneously with glucose, rather than sequentially. Based upon fermentations of sugar mixtures, a catabolite-repression mutant of ethanologenic E. coli is expected to provide more efficient fermentation of hemicellulose hydrolysates by allowing direct utilization of pentoses. PMID- 11499919 TI - Optimisation of media and cultivation conditions for L(+)(S)-lactic acid production by Lactobacillus casei NRRL B-441. AB - Process variables and concentration of carbon in media were optimised for lactic acid production by Lactobacillus casei NRRL B-441. Lactic acid yield was inversely proportional to initial glucose concentration within the experimental area (80-160 g l(-1)). The highest lactic acid concentration in batch fermentation, 118.6 g l(-1), was obtained with 160 g 1(-1) glucose. The maximum volumetric productivity, 4.4 g 1(-1) h(-1) at 15 h, was achieved at an initial glucose concentration of 100 g l(-1). Similar lactic acid concentrations were reached with a fedbatch approach using growing cells, in which case the fermentation time was much shorter. Statistical experimental design and response surface methodology were used for optimising the process variables. The temperature and pH optima for lactic acid production were 35 degrees C, pH 6.3. Malt sprout extract supplemented with yeast extract (4 g l(-1)) appeared to be an economical alternative to yeast extract alone (22 g l(-1)) although the fermentation time was a little longer. The results demonstrated both the separation of the growth and lactic acid production phases and lactic acid production by non-growing cells without any nutrient supplements. Resting L. casei cells converted 120 g l(-1) glucose to lactic acid with 100% yield and a maximum volumetric productivity of 3.5 g l(-1) h(-1). PMID- 11499920 TI - Comparative study of the relationship between monomer structure and reactivity for two polyhydroxyalkanoate synthases. AB - Using organically synthesized hydroxyalkanoate coenzyme A thioesters, the activities of two short-chain polyhydroxalkanoate (PHA) synthases were investigated--Ralstonia eutropha PHA synthase (a type I PHA synthase) and Ectothiorhodospira shaposhnikovii PHA synthase (a type III synthase). The results indicate that the two synthases have similar activities towards most of the monomers tested. 3-Hydroxybutyryl CoA was found to be the most efficient substrate for both synthases. Changes in the side-chain length of the monomers affect monomer reactivity, with shortening of the side-chain length having the more severe effect. Hydrophobicity in the side chain appears to play an important role in the catalytic reaction. The configuration and the position of the hydroxyl group also affect the reactivity of a monomer. Monomers with the [S] configuration can not be recognized by either synthase. Moving the hydroxyl group from the beta carbon to the alpha carbon has a much more severe effect on the reactivity of the monomer than moving the hydroxyl group to the gamma carbon. The results demonstrate that the in vitro system can be used to prepare entirely novel polymers that may not be obtainable from living cells because of metabolic restrictions. PMID- 11499921 TI - Characterization of a chitinase and an endo-beta-1,3-glucanase from Trichoderma harzianum Rifai T24 involved in control of the phytopathogen Sclerotium rolfsii. AB - Of 24 Trichoderma isolates, T harzianum Rifai (T24) showed a potential for control of the phytopathogenic basidiomycete Sclerotium rolfsii. When T24 was grown on different carbon sources, growth inhibition of S. rolfsii by the T24 culture filtrate correlated with the activity of extracellular chitinase and beta 1,3-glucanase. The 43-kilodalton (kDa) chitinase and the 74-kDa beta-1,3 glucanase were purified from the T24 culture filtrate in two and three steps, respectively, using ammonium sulphate precipitation followed by hydrophobic interaction chromatography (phenyl-Sepharose) and gel filtration (beta-1,3 glucanase). Km and Kcat were 3.8 g l(-1) and 0.71 s(-1) for the chitinase (chitin) and 1.1 g(-1) and 52 s(-1) for the beta-1,3-glucanase (laminarin). The chitinase showed higher activity on chitin than on less-acetylated substrate analogues (chitosan), while the beta-1,3-glucanase was specific for beta-1,3 linkages in polysaccharides. Both enzymes were stable at 30 degrees C, while at 60 degrees C the chitinase and the beta-1,3-glucanase were rapidly inactivated, showing half-lives of 15 and 20 min, respectively. The enzymes inhibited growth of S. rolfsii in an additive manner showing a promising ED50 (50% effective dose) value of 2.7 microg/ml. PMID- 11499922 TI - Construction of an expression vector for propionibacteria and its use in production of 5-aminolevulinic acid by Propionibacterium freudenreichii. AB - Several promoters from Propionibacterium freudenreichii subsp. shermanii were isolated using a promoter probe vector, pCVE1, containing the Streptomyces cholesterol oxidase gene (choA) as a reporter gene. Three of four promoters isolated exhibiting a strong activity in Escherichia coli also expressed a strong activity in P. freudenreichii subsp. shermanii IFO12426. Using two promoters with a strong activity and a previously constructed shuttle vector, pPK705, shuttling between E. coli and Propionibacterium. we constructed expression vectors for propionibacteria. To overproduce 5-aminolevulinic acid (ALA), which is the first intermediate in the synthesis of porphyrins, the ALA synthase gene (hemA) from Rhodobacter sphaeroides was recombined with the expression vectors. The activity of ALA synthase in the recombinant P freudenreichii subsp. shermanii increased about 70-fold that in the strain without a vector. The recombinant Propionibacterium produced ALA at a maximum concentration of 8.6 mM in the absence of levulinic acid, an inhibitor of ALA dehydratase, with 1% glucose as a carbon source. The recombinant P. freudenreichii accumulated 18.8 mmol/g cells ALA in the presence of 1 mM levulinic acid and 30 mM glycine. The construction of an efficient expression vector will facilitate genetic studies of a vitamin B12 producer, Propionibacterium. PMID- 11499923 TI - Efficiency of delivery of DNA to cells by bovine papillomavirus type-1 L1/L2 pseudovirions. AB - To investigate the efficiency of encapsidation of plasmid by papillomavirus virus like particles (PV VLPs), and the infectivity of the resultant PV pseudovirions, Cos-1 cells were transfected with an 8-kb plasmid incorporating a green fluorescent protein (GFP) reporter gene (pGSV), and infected with bovine PV (BPV 1) L1/L2 recombinant vaccinia virus to produce BPV1 pseudovirions. Approximately 1 in 1.5 x 10(4) of dense (1.35 g/ml) PV pseudovirions and 0.3 in 10(4) of less dense (1.29 g/ml) pseudovirions packaged an intact pGSV plasmid. The majority (>75%) of packaged plasmids contained deletions, and the deletions affected all tested genes. After exposure of Cos-1 cells to BPV-1 pseudovirions at an MOI of 40,000:1, 6% of cells expressed GFP, giving a calculated efficiency of delivery of the pGSV plasmid, by pseudovirions which had packaged an intact plasmid, of approximately 5%. Plasmid delivery was not effected by purified pGSV plasmid, was blocked by antiserum against BPV-1, and was not blocked by DNase treatment of pseudovirions, confirming that delivery was mediated by DNA within the pseudovirion. We conclude that a major limitation to the use of PV pseudovirions as a gene delivery system is that intact plasmid DNA is not efficiently selected for packaging by VLPs in cell-based pseudovirions production systems. PMID- 11499924 TI - Expression of mouse anticreatine kinase (MAK33) monoclonal antibody in the yeast Hansenula polymorpha. AB - The methylotrophic yeast Hansenula polymorpha HM1-39 (ura 3 and leu 2) was used as a host strain for the expression of the Fab fragment of the MAK33 monoclonal antibody. The MAK33 antibody reacts specifically with creatine kinase-M. The cDNA of kappa and gamma chains were inserted between the FMD or MOX promoter and the MOX terminator within the expression plasmids. In addition, the secretion signal sequence of the mating factor-alpha (prepro segment) and a fragment from glucoamylase with its secretion signal peptide, were also inserted in the expression plasmids for efficient secretion and production of the MAK33 monoclonal antibody. The co-expression of kappa and gamma chains was achieved by double transformation with kappa and then with gamma chain-expressing plasmids. The cells of H. polymorpha HM1-39 showed high mitotic stability and both uracil+ and leucine+ phenotypic stability after double transformation. Northern analysis showed a high rate of transcription of either kappa or gamma chain mRNA but not both, when the cells were grown in an induction medium. Protein analysis of double-transformed cells showed the monomers of the MAK33 antibody (kappa and gamma chains) were not assembled into a heterodimeric functional form. The expressed proteins of light and heavy chains represent about 11-12% of total cell protein and are found more inside than outside the cell. The expressed monomers show antigen-binding affinity in the Ouchterlony diffusion test; and the binding activity exhibited by cell-free extract was more than that of the cell culture supernatant. PMID- 11499925 TI - Expression, processing and high level secretion of a virus toxin in fission yeast. AB - The virally encoded K28 toxin of Saccharomyces cerevisiae kills sensitive yeast cells in a multi-step receptor-mediated fashion by cell cycle arrest and inhibition of DNA synthesis. In vivo, the toxin is translated as a 38 kDa preprotoxin (pptox) which is enzymatically processed to the biologically active alpha/beta heterodimer during passage through the yeast secretory pathway. Here, we demonstrate that Schizosaccharomyces pombe, a yeast from which no natural toxin-secreting killer strains are known, is perfectly capable of expressing a killer phenotype. Episomal as well as integrating K28 pptox gene cassettes were constructed that allowed a tightly thiamine-regulated killer phenotype expression under transcriptional control of the Sch. pombe nmt1 promotor. Northern analysis of the toxin-coding transcript as well as Western analysis of the secreted toxin indicated that fission yeast is capable of expressing a correctly processed and fully functional virus toxin. Moreover, toxin secretion in recombinant Sch. pombe was at least ten-fold higher than in any natural and/or recombinant Sac. cerevisiae killer strain, indicating that pptox-derived vectors might be attractive in the fast growing field of heterologous protein expression and secretion in yeast. PMID- 11499926 TI - Fuel ethanol production from lignocellulose: a challenge for metabolic engineering and process integration. AB - With industrial development growing rapidly, there is a need for environmentally sustainable energy sources. Bioethanol (ethanol from biomass) is an attractive, sustainable energy source to fuel transportation. Based on the premise that fuel bioethanol can contribute to a cleaner environment and with the implementation of environmental protection laws in many countries, demand for this fuel is increasing. Efficient ethanol production processes and cheap substrates are needed. Current ethanol production processes using crops such as sugar cane and corn are well-established; however, utilization of a cheaper substrate such as lignocellulose could make bioethanol more competitive with fossil fuel. The processing and utilization of this substrate is complex, differing in many aspects from crop-based ethanol production. One important requirement is an efficient microorganism able to ferment a variety of sugars (pentoses, and hexoses) as well as to tolerate stress conditions. Through metabolic engineering, bacterial and yeast strains have been constructed which feature traits that are advantageous for ethanol production using lignocellulose sugars. After several rounds of modification/evaluation/modification, three main microbial platforms, Saccharomyces cerevisiae, Zymomonas mobilis, and Escherichia coli, have emerged and they have performed well in pilot studies. While there are ongoing efforts to further enhance their properties, improvement of the fermentation process is just one of several factors-that needs to be fully optimized and integrated to generate a competitive lignocellulose ethanol plant. PMID- 11499927 TI - Identification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis. AB - The functional importance of a conserved region in a novel chitosanase from Bacillus sp. CK4 was investigated. Each of the three carboxylic amino acid residues (Glu-50, Glu-62, and Asp-66) was changed to Asp and Gln or Asn and Glu by site-directed mutagenesis, respectively. The Asp-66-->Asn and Asp-66-->Glu mutation remarkably decreased kinetic parameters such as Vmax and kcat to approximately 1/1,000 those of the wild-type enzyme, indicating that the Asp-66 residue was essential for catalysis. The thermostable chitosanase contains three Cys residues at positions 49, 72, and 211. The Cys-49-->Ser/Tyr and Cys-72- >Ser/Tyr mutant enzymes were as stable to thermal inactivation and denaturating agents as the wild-type enzyme. However, the half-life of the Cys-211-->Ser/Tyr mutant enzyme was less than 10 min at 80 degrees C, while that of the wild-type enzyme was about 90 min. Moreover, the residual activity of Cys-211-->Ser/Tyr enzyme was substantially decreased by 8 M urea; and it lost all catalytic activity in 40% ethanol. These results show that the substitution of Cys with any amino acid residues at position 211 seems to affect the conformational stability of the chitosanase. PMID- 11499928 TI - Polyketide synthase genes in insect- and nematode-associated fungi. AB - Production of polyketides is accomplished through complex enzymes known as polyketide synthases (PKS); these enzymes have highly conserved domains that might be useful in screens for PKSs in diverse groups of organisms. A degenerate PCR-based approach was used to amplify PKS fragments of the ketosynthase domain from genomic DNA of a group of insect- and nematode-associated fungi. Of 157 isolates (representing 73 genera and 144 species) screened, 92 isolates generated PCR products of predicted size (approximately 300 bp). The ability to detect PKS domains was a function of the number of different primer pairs employed in the screen. Cloning and sequencing revealed that 66 isolates had at least one unique PKS sequence; ten members of this set contained multiple PKS fragments, for a total of 76 unique PKS fragments. Since PKS genes appear to be widespread among fungi, a PCR-based screening system appears to be an efficient, directed means to identify organisms having the potential to produce polyketides. PMID- 11499930 TI - Microbial desulfurization of alkylated dibenzothiophene and alkylated benzothiophene by recombinant Rhodococcus sp. strain T09. AB - The dibenzothiophene (DBT) desulfurizing operon, dsz, was introduced into various benzothiophene (BT)-desulfurizing bacteria using a Rhodococcus-E. coli shuttle vector. Of the tested recombinant bacteria, only those from Rhodococcus sp. strain T09 grew with both DBT and BT as the sole sulfur source. These recombinant cells desulfurized not only alkylated BTs, but also various alkylated DBTs, producing alkylated hydroxybiphenyls as the desulfurized products. Recombinant strain T09 also desulfurized alkylated DBT in an oil-water, two-phase resting cell reaction. The dsz operon had the same desulfurizing activity when inserted into the vector in either orientation, indicating that the promoter region of the operon was functional in strain T09. PMID- 11499929 TI - The physiological effects and metabolic alterations caused by the expression of Rhizobium etli pyruvate carboxylase in Escherichia coli. AB - Oxaloacetate (OAA) plays an important role in the tricarboxylic acid cycle and for the biosynthesis of a variety of cellular compounds. Some microorganisms, such as Rhizobium etli and Corynebacterium glutamicum, are able to synthesize OAA during growth on glucose via either of the enzymes pyruvate carboxylase (PYC) or phosphoenolpyruvate carboxylase (PPC). Other microorganisms, including Escherichia coli, synthesize OAA during growth on glucose only via PPC because they lack PYC. In this study we have examined the effect that the R. etli PYC has on the physiology of E. coli. The expressed R. etli PYC was biotinylated by the native biotin holoenzyme synthase of E. coli and displayed kinetic properties similar to those reported for alpha4 PYC enzymes from other sources. R. etli PYC was able to restore the growth of an E. coli ppc null mutant in minimal glucose medium, and PYC expression caused increased carbon flow towards OAA in wild-type E. coli cells without affecting the glucose uptake rate or the growth rate. During aerobic glucose metabolism, expression of PYC resulted in a 56% increase in biomass yield and a 43% decrease in acetate yield. During anaerobic glucose metabolism, expression of PYC caused a 2.7-fold increase in succinate concentration, making it the major product by mass. The increase in succinate came mainly at the expense of lactate formation. However, in a mutant lacking lactate dehydrogenase activity, expression of PYC resulted in only a 1.7-fold increase in succinate concentration. The decreased enhancement of succinate formation in the /dh mutant was hypothesized to be due to accumulation of pyruvate and NADH, metabolites that affect the interconversion of the active and inactive form of the enzyme pyruvate formate-lyase. PMID- 11499931 TI - Genetic transformation of Trametes versicolor to phleomycin resistance with the dominant selectable marker shble. AB - We have developed a stable, DNA-mediated transformation system for the white-rot basidiomycete Trametes versicolor based on the dominant selectable marker shble (phleomycin resistance). We employed a vector containing the selectable marker under control of expression sequences from the basidiomycete Schizophyllum commune and a polyethylene glycol/ CaCl2 protoplast-fusion technique to introduce the transforming DNA. This transformation system generated stable phleomycin resistant transformants at a frequency of four to seven transformants/microg of transforming DNA. PMID- 11499932 TI - Cloning, expression, and carbon catabolite repression of the bamM gene encoding beta-amylase of Bacillus megaterium DSM319. AB - The bamM gene from Bacillus megaterium DSM319 encoding an extracellular beta amylase was isolated and completely sequenced. Chromosomal inactivation by deletion mutagenesis resulted in total loss of amylolytic activity, indicative of a single starch-degrading enzyme. Functional characterization of the expressed protein revealed a maltogenic enzyme exhibiting optimal activities at pH 7.5 and 50 degrees C. Amylase expression is subject to catabolite repression by glucose. A putative cis-acting catabolite-responsive element (CRE) was identified; it is located within the bamM coding region, matching the position of the predicted signal peptide processing site. Base substitutions introduced by site-directed mutagenesis within the bamM-CRE--retaining unchanged the amino acid sequence- provoked a remarkable relief from carbon catabolite repression (CCR), thereby proving functionality of the CRE for CCR. PMID- 11499933 TI - Functional patterns and temperature response of cellulose-fermenting microbial cultures containing different methanogenic communities. AB - The effect of microbial composition on the methanogenic degradation of cellulose was studied using two lines of anaerobic cellulose-fermenting methanogenic microbial cultures at two different temperatures: that at 15 degrees C being dominated by Methanosaeta and that at 30 degrees C by Methanosarcina. In both cultures, CH4 production and acetate consumption were completely inhibited by either 2-bromoethanesulfonate or chloroform, whereas H2 consumption was only inhibited by chloroform, suggesting that homoacetogens utilized H2 concomitantly with methanogens. Hydrogen was the intermediate that was consumed first, while acetate continued to accumulate. At 15 degrees C, acetoclastic methanogenesis smoothly followed H2-dependent CH4 production. Fluorescence in situ hybridization showed that populations of Methanosaeta steadily increased with time from 5 to 25% of total cell counts. At 30 degrees C, two phases of CH4 production were obtained, with acetate consumed after the abrupt increase of Methanosarcina from 0 to 45% of total cell counts. Whereas populations of Methanosaeta were able to adapt after transfer from 15 to 30 degrees C, those of Methanosarcina were not, irrespective of during which phase the cultures were transferred from 30 degrees C to 15 degrees C. Our results thus show that the community structure of methanogens indeed affects the function of a cellulose-fermenting community with respect to temperature response. PMID- 11499934 TI - The influence of supplemental components in nutrient medium on chitosan formation by the fungus Absidia orchidis. AB - Chitosan, a derivative of chitin, is a natural component of some fungus cell walls. It is formed by the complex action of chitin synthase and chitin deacetylase. The in vitro activity of these two enzymes is known to be influenced by several factors. We investigated the influence of ferrous ions, manganese ions, cobalt ions, trypsin, and chitin, as individual supplements to the nutrient medium, on the in vivo activity of chitin synthase and chitin deacetylase to form chitosan in the fungus Absidia orchidis. Manganese and ferrous ions gave the most significant results. These ions increase chitosan yields through an increase in biomass production rather than an increase of chitosan content in cell walls. Manganese and ferrous ions lowered the activity of chitin deacetylase; however, their influence on the activity of chitin synthase was more complex. The effects of trypsin and chitin on biomass and cell wall chitosan content were negligible, while cobalt ions completely inhibited the growth of fungi. PMID- 11499936 TI - Mycelial pellet intrastructure and visualization of mycelia and intracellular lipid in a culture of Mortierella alpina. AB - The intrastructure of mycelial pellets of Mortierella alpina, which accumulate fatty acids in mycelia, was visualized following labeling with fluorescein isothiocyanate (FITC) and Nile red using fluorescence microscopy. The pellet was an ellipse shape, but its intrastructure was shaped as a doughnut with a cave inside. Using three-dimensional image analysis, it was shown that the lipid was produced on the edge of the pellet, which corresponded to the area where the mycelial density was high. The cavity ratio of the pellet section was determined on the basis of the FITC fluorescence intensity, and in the early culture stage remained at 0.2 in a 10-kl fermentor culture, but finally increased to 0.35. Mycelial pellet volume paralleled the cavity ratio. Application of the technique used here allows analysis of the intrastructure of fungal pellets and new types of fungal biological study. PMID- 11499935 TI - Enhanced formation of laccase activity by the white-rot fungus Trametes pubescens in the presence of copper. AB - The white-rot fungus Trametes pubescens MB 89 has been identified as an outstanding, although not-yet-described, producer of the industrially important enzyme laccase. Extracellular laccase formation could be greatly stimulated by the addition of Cu(II) to a simple, glucose-based culture medium. Using optimum Cu concentrations (1.5-2.0 mM), maximum values for laccase activity of approximately 65 U/ml were obtained. The synthesis of the laccase protein depended on the presence of Cu in the medium as shown by Western blot analysis. Copper had to be supplemented during the exponential phase of growth for its maximal effect; addition during the stationary phase, during which laccase activity is predominantly formed, resulted in markedly reduced laccase productivity. As was shown by X-ray microanalysis of T pubescens mycelia obtained from a laboratory fermentation, Cu was rapidly taken up by the fungal biomass. A possible explanation for this stimulatory effect of Cu on laccase biosynthesis could be a role for this enzyme activity in melanin synthesis. The stimulatory effect of Cu on laccase synthesis was also effective for several other basidiomycetes and hence could be used as a simple method to boost the production of this enzyme. PMID- 11499937 TI - Accumulation of 2,5-dimethoxy-1,4-benzoquinone in suspension cultures of Panax ginseng by a fungal elicitor preparation and a yeast elicitor preparation. AB - Suspension cultures of Panax ginseng C.A. Meyer (Araliaceae) were treated with either an elicitor preparation from the culture broth of the phytopathogenic hyphomycete Botrytis cinerea or a yeast elicitor preparation, and the accumulation of a new compound, which was not detected in non-elicited cultures, was observed. The accumulated compound was isolated and shown to be 2,5-dimethoxy 1,4-benzoquinone by 1H-NMR, 13C-NMR and electron ionization (EI) mass spectra. While it is well known that this compound shows antibacterial activity against Staphylococcus aureus, its presence in ginseng root has not been reported to date. Levels of the compound in the media increased rapidly, reaching a maximum level of 65.10 +/- 4.96 microg/g fresh weight at approximately 12 h after treatment with the yeast elicitor preparation. The maximal level of the compound in medium from the culture treated with an elicitor preparation from the culture broth of B. cinerea was 46.13 +/- 10.42 microg/g fresh weight after 24 h of incubation. PMID- 11499938 TI - Antimicrobial activity of argon fluoride (ArF) excimer laser on gram-negative bacteria. AB - The objective of this study was to evaluate the antibacterial activity of argon fluoride (ArF) excimer laser radiation on clinically important strains of gram negative bacteria. The antibacterial activity of ArF excimer laser radiation was evaluated on two Acinetobacter baumannii, one Enterobacter cloacae, three Escherichia coli, two Helicobacter pylori, one Klebsiella pneumoniae and two Pseudomonas aeruginosa strains. The strains were isolated from clinical specimens and typed by the usual biochemical procedures. Square agar plates of 12 x 12 cm were divided into rectangular (2 x 3 cm) regions and spread with 0.5x 10(4) colony forming units (CFU)/ml of bacterial suspension. The excess liquid was removed and the plates were allowed to dry for 30 min. A total of 96 rectangular (2x3 cm) regions were used for each strain, in order to test an equal number of laser parameters. Each rectangular region was irradiated with different laser parameters, using a 193 nm ArF excimer laser, linked with a simple Galilean afocal system and a rectangular diaphragm of the same dimensions as the original laser beam cross-section, at a distance of 10 cm from the irradiated surface. This system was used in order to keep the laser pulse energy under 80 mJ and to cut-out the non-transverse electromagnetic mode branches of the laser beam. We then studied the bacterial survival ratio versus the number of laser pulses, the repetition frequency and the total laser beam fluence. Our results showed that the total laser beam fluence was the most important parameter to consider in evaluating the bactericidal effect of ArF excimer laser radiation. A critical value of the total fluence was determined for each strain, such that, for laser beam fluences greater than this critical value, no colonies appeared to survive while, for laser fluences less than this critical value, the survival ratio did not exceed 2 x 10(7) CFU (2 x 10(-5)%). These critical values were found to vary between 8 J/cm2 and 16 J/cm2 for the bacterial species studied. Under these conditions, ArF laser irradiation is promising for the sterilisation of hard surfaces and for in situ application. PMID- 11499939 TI - The effect of osmotic pressure on the membrane fluidity of Saccharomyces cerevisiae at different physiological temperatures. AB - Membrane fluidity in whole cells of Saccharomyces cerevisiae W303-1A was estimated from fluorescence polarization measurements using the membrane probe, 1,6-diphenyl-1,3,5-hexatriene, over a wide range of temperatures (6-35 degrees C) and at seven levels of osmotic pressure between 1.38 MPa and 133.1 MPa. An increase in phase transition temperatures was observed with increasing osmotic pressure. At 1.38 MPa, a phase transition temperature of 12 +/- 2 degrees C was observed, which increased to 17 +/- 4 degrees C at 43.7 MPa, 21+/- 7 degrees C at 61.8 MPa, and 24 +/- 9 degrees C at an osmotic pressure of 133.1 MPa. From these results we infer that, with increases in osmotic pressure, the change in phospholipid conformation occurs over a larger temperature range. These results allow the representation of membrane fluidity as a function of temperature and osmotic pressure. Osmotic shocks were applied at two levels of osmotic pressure and at nine temperatures, in order to relate membrane conformation to cell viability. PMID- 11499940 TI - Introduction of green fluorescent protein gene into phenol-degrading Alcaligenes faecalis cells and their monitoring in phenol-contaminated soil. AB - Alcaligenesfaecalis (CCT 7145) was isolated from an Amazonian soil sample after an enrichment process to select for phenol-degrading microorganisms. The isolate was labeled with the green fluorescent protein (gfp) gene. The gfp-transformed cells were easily detected using a hand-held UV transilluminator and their taxonomy was confirmed by 16S rRNA sequencing. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that the gfp gene was integrated into the chromosome. The addition of the gfp marker did not affect phenol degradation ability compared with the wild-type. Both, wild-type and gfp-marked A. faecalis cells encapsulated in alginate, tolerated 1,700 microg ml(-1) phenol in liquid medium compared with 1,100 microg ml(-1) phenol for free cells. 14C-Phenol mineralization in soil microcosms was also enhanced by inoculation with encapsulated cells. Survival of gfp-marked cells in phenol-contaminated soil over 22 days was determined from plate counts using an epifluorescence microscope. PMID- 11499941 TI - A simple mediatorless amperometric method using the cyanobacterium Synechococcus leopoliensis for the detection of phytotoxic pollutants. AB - The unicellular cyanobacterium Synechoccocus leopoliensis is used in a micro electrochemical cell to generate photocurrents. The photocurrent is dependent on photosynthetic electron transport and is mediated by hydrogen peroxide formation following the reduction of oxygen on the acceptor side of photosystem I. This is the first known application of cyanobacteria in an electrochemical device where no artificial electroactive mediator is needed. The potential for the development of this micro-electrochemical cell for the detection of phytotoxic pollutants, such as herbicides and toxic metal cations, using the photosynthetic system of the cyanobacteria without interference from added electron acceptor is discussed. PMID- 11499942 TI - Aerobic degradation of mixtures of tetrachloroethylene, trichloroethylene, dichloroethylenes, and vinyl chloride by toluene-o-xylene monooxygenase of Pseudomonas stutzeri OX1. AB - A recombinant strain of Escherichia coli (JM109/pBZ1260) expressing constitutively toluene-o-xylene monooxygenase (ToMO) of Pseudomonas stutzeri OX1 degraded binary mixtures (100 microM each) of tetrachloroethylene (PCE) with either trichloroethylene (TCE), 1,1-dichloroethylene (1,1-DCE), cis dichloroethylene (cis-DCE), trans-1,2-dichloroethylene (trans-DCE), or vinyl chloride (VC). PCE degradation was 8-20% for these binary mixtures, while TCE and trans-DCE with PCE were degraded at 19%, 1,1-DCE at 37%, cis-DCE at 97%, and VC at 27%. The host P. stutzeri OXI was also found to degrade binary mixtures of PCE/TCE, PCE/cis-DCE, and PCE/VC when induced with toluene. Degradation of quaternary mixtures of PCE/TCE/trans-DCE/VC and PCE/TCE/cis-DCE/VC by JM109/pBZ1260 were also investigated as well as mixtures of PCE/TCE/trans-DCE/1,1 DCE/cis-DCE/VC; when all the chlorinated compounds were present, the best degradation occurred with 24-51% removal of each. For these degradation reactions, 39-85% of the stoichiometric chloride expected from complete degradation of the chlorinated ethenes was detected. The time course of PCE/TCE/1,1-DCE degradation was also measured for a mixture of 8, 17, and 6 microM, respectively; initial degradation rates were 0.015, 0.023. and 0.029 nmol/min x mg protein, respectively. This indicates that for the first time an aerobic enzyme can degrade mixtures of all chlorinated ethenes, including the once--so it was believed-completely recalcitrant PCE. PMID- 11499943 TI - Isolation and characterization of a thermotolerant bacterium Ralstonia sp. strain PHS1 that degrades benzene, toluene, ethylbenzene, and o-xylene. AB - A thermotolerant bacterium, designated as PHS1, was isolated from a hot spring in Pohang, Korea, on the basis of its ability to grow on benzene, toluene, ethylbenzene, and xylenes (BTEX) as a sole carbon source. Strain PHS1 is a gram negative, rod-shaped aerobe and grows optimally at 42 degrees C and pH 7.2. According to 16 S rDNA analysis, strain PHS1 showed highest similarity to Ralstonia eutropha (previously named Alcaligenes eutrophus). Unlike its closest known Ralstonia species, however, strain PHS1 was able to utilize toluene, ethylbenzene, o-xylene, and both m- and o-cresol. The degradation of o-xylene by strain PHS1 is particularly important, since o-xylene is a compound of considerable environmental interest, owing to its recalcitrance; and very few microorganisms have been reported to utilize o-xylene as a sole carbon source. It was found that strain PHS1 transformed o-xylene to 2,3-dimethylphenol through direct oxygenation of the aromatic ring. The unique properties of strain PHS1, such as thermotolerance and the ability to degrade o-xylene, may have important implications for the treatment of BTEX-contaminated industrial effluents. PMID- 11499944 TI - Bioaccumulation of mercury from wastewater by genetically engineered Escherichia coli. AB - Genetically engineered E. coli, which express both a Hg2+ transport system and metallothionein, were tested for their ability to remove mercury from wastewater. The wastewater contained more than ten different ions, including 2.58 mg/l mercury, and its pH was 9.6. Mercury uptake was faster from the wastewater than from distilled water, probably because of the higher ionic strength, as the high pH had little effect on mercury accumulation. EDTA also stimulated mercury uptake rather than inhibiting it. A hollow-fiber bioreactor was used to retain induced cells for continuous mercury uptake. The cells removed more than 99% of the mercury in the wastewater and the final amount of mercury accumulated was 26.8 mg/g cell dry weight, while none of the other ions were removed from the water. These results indicated that the induced cells had a high affinity and specificity for mercury. PMID- 11499945 TI - Methane fermentation of coastal mud sediment by a two-stage upflow anaerobic sludge blanket (UASB) reactor system. AB - The removal of organic matter from a coastal mud sediment was carried out by a methane fermentation process under anaerobic conditions. In a batch acidogenic fermentation, the addition of vitamins containing thiamine, nicotinic acid and biotin dramatically enhanced acetate production from the mud sediment (200 g wet wt l(-1) artificial sea water), yielding 77 mM acetate after 6 days, which corresponded to 77% of the organic matter in the mud sediment, measured on the basis of chemical oxygen demand. Thereafter, the two-fold diluted, post acidogenic fermentation liquor (PAF liquor) was continuously treated at 2.4x original dilution rate day(-1) for 30 days, using an upflow anaerobic sludge blanket methanogenic reactor containing the acclimated methanogenic sludge from the mud sediment. Acetate, 42 mM in the PAF liquor, was converted to methane at a maximum methane production rate of 96 mmol l(-1) day(-1); and 87.5% of the acetate and 88.7% of the total organic carbon in the PAF liquor were removed. Moreover, an efficient treatment of the mud sediment was carried out by a semi continuous, two-stage reactor system, where the culture broth was circulated between acidogenic and methanogenic reactors. This two-stage reactor system gave a stable operation at 4-day intervals for one treatment period, yielding 112 mmol methane from the wet mud in the PAF liquor (278 g l(-1)). PMID- 11499946 TI - The use of alternative technologies to develop malolactic fermentation in wine. AB - The development of the malolactic fermentation, bioconversion of L-malic acid to L-lactic acid, is a difficult and time-consuming process that does not always proceed favorably under the natural conditions of wine. Traditional fermentations are used worldwide to produce high-quality wines, although delay or failure is not an unusual outcome. During recent years several technologies have been proposed to induce biological deacidification of wines by using malolactic bacteria, principally Oenococcus oeni and Lactobacillus sp. These alternative technologies usually involve the use of high densities of cells or enzymes, free or immobilized onto different matrices. Immobilization materials, several types of bioreactors, and the properties of many specific systems are discussed in this review. PMID- 11499947 TI - Application of rRNA-targeted oligonucleotide probes in biotechnology. AB - Ribosomal RNA-targeted oligonucleotide probes have become valuable tools for the detection of microorganisms involved in important biotechnological processes. Microorganisms which are of major importance for processes such as wastewater treatment, microbial leaching or methane production can be detected and quantified in situ within a complex microbial community. For certain processes, such as nitrification or biological phosphate removal, new microorganisms have become the focus of interest and have led to an improved understanding of these bioremediation techniques. Hybridization techniques have become fast and reliable alternatives to conventional cultivation techniques in the food industry as a control method for starter cultures for fermentation processes or product control. Recent analytical tools such as flow cytometry and digital image processing have improved the efficiency of these techniques. This review is intended to present a summary of methodological aspects of rRNA-based hybridization techniques and their application in biotechnology. PMID- 11499948 TI - The fungal biocontrol agent Coniothyrium minitans: production by solid-state fermentation, application and marketing. AB - Biological control agents (BCAs) are potential alternatives for the chemical fungicides presently used in agriculture to fight plant diseases. Coniothyrium minitans is an example of a promising fungal BCA. It is a naturally occurring parasite of the fungus Sclerotinia sclerotiorum, a wide-spread pathogen which substantially reduces the yield of many crops. This review describes, exemplified by C. minitans, the studies that need to be carried out before a fungal BCA is successfully introduced into the market. The main aspects considered are the biology of C. minitans, the development of a product by mass production of spores using solid-state fermentation technology, its biocontrol activity and marketing of the final product. PMID- 11499949 TI - Basic and applied aspects in the microbial degradation of azo dyes. AB - Azo dyes are the most important group of synthetic colorants. They are generally considered as xenobiotic compounds that are very recalcitrant against biodegradative processes. Nevertheless, during the last few years it has been demonstrated that several microorganisms are able, under certain environmental conditions, to transform azo dyes to non-colored products or even to completely mineralize them. Thus, various lignolytic fungi were shown to decolorize azo dyes using ligninases, manganese peroxidases or laccases. For some model dyes, the degradative pathways have been investigated and a true mineralization to carbon dioxide has been shown. The bacterial metabolism of azo dyes is initiated in most cases by a reductive cleavage of the azo bond, which results in the formation of (usually colorless) amines. These reductive processes have been described for some aerobic bacteria, which can grow with (rather simple) azo compounds. These specifically adapted microorganisms synthesize true azoreductases, which reductively cleave the azo group in the presence of molecular oxygen. Much more common is the reductive cleavage of azo dyes under anaerobic conditions. These reactions usually occur with rather low specific activities but are extremely unspecific with regard to the organisms involved and the dyes converted. In these unspecific anaerobic processes, low-molecular weight redox mediators (e.g. flavins or quinones) which are enzymatically reduced by the cells (or chemically by bulk reductants in the environment) are very often involved. These reduced mediator compounds reduce the azo group in a purely chemical reaction. The (sulfonated) amines that are formed in the course of these reactions may be degraded aerobically. Therefore, several (laboratory-scale) continuous anaerobic/aerobic processes for the treatment of wastewaters containing azo dyes have recently been described. PMID- 11499950 TI - Microbial decolourisation and degradation of textile dyes. AB - Dyes and dyestuffs find use in a wide range of industries but are of primary importance to textile manufacturing. Wastewater from the textile industry can contain a variety of polluting substances including dyes. Increasingly, environmental legislation is being imposed to control the release of dyes, in particular azo-based compounds, into the environment. The ability of microorganisms to decolourise and metabolise dyes has long been known, and the use of bioremediation based technologies for treating textile wastewater has attracted interest. Within this review, we investigate the mechanisms by which diverse categories of microorganisms, such as the white-rot fungi and anaerobic bacterial consortia, bring about the degradation of dyestuffs. PMID- 11499951 TI - Changes in morphology of Paecilomyces japonica and their effect on broth rheology during production of exo-biopolymers. AB - The influence of Paecilomyces japonica pellet morphology on fermentation broth rheology and exobiopolymer production was investigated in a 5-1 jar fermenter. Rapid formation of pellets was observed after the first day of fermentation; and these slowly increased in size and roughness. This, together with the increase in biomass concentration, altered the transport characteristics and broth rheology towards a pseudoplastic nature which, in turn, influenced cell growth and exo biopolymer production. At mild agitation, high aeration and optimum substrate concentration, pellets were the most predominant morphological form, compared with free mycelia. The broth rheology showed pseudoplastic behavior; and the fungal morphology was closely related to the rheological properties. PMID- 11499952 TI - Pharmaceutically relevant metabolites from lichens. AB - Lichen metabolites exert a wide variety of biological actions including antibiotic, antimycobacterial, antiviral, antiinflammatory, analgesic, antipyretic, antiproliferative and cytotoxic effects. Even though these manifold activities of lichen metabolites have now been recognized, their therapeutic potential has not yet been fully explored and thus remains pharmaceutically unexploited. In this mini-review, particular attention is paid to the most common classes of small-molecule constituents of lichens, from both the chemical viewpoint and with regard to possible therapeutic implications. In particular, aliphatic acids, pulvinic acid derivatives, depsides and depsidones, dibenzofuans, anthraquinones, naphthoquinones as well as epidithiopiperazinediones are described. An improved access to these lichen substances in drug discovery high-throughput screening programs will provide impetus for identifying novel lead-compounds with therapeutic potential and poses new challenges for medicinal chemistry. PMID- 11499953 TI - Changes of in vivo fluxes through central metabolic pathways during the production of nystatin by Streptomyces noursei in batch culture. AB - The central carbon metabolism of the nystatin-producing strain Streptomyces noursei ATCC 11455 was evaluated by 13C-labelling experiments. A batch fermentation was examined during the idiophase by GC-MS measurements of the labelling patterns of amino acids in the biomass. The labelling patterns of the amino acids and calculated fluxes of the central metabolism showed that changes in the primary and secondary metabolisms occurred simultaneously. Changes in the profiles for the integrated fluxes showed a decreased flux through the pentose phosphate pathway and an increased flux in the tricarboxylic acid cycle relative to the glucose uptake rate when the culture entered a phase with reduced specific growth rate and enhanced nystatin yield. The flux through the pentose phosphate pathway seemed to be adjusted according to the NADPH requirement during the different phases of the batch fermentation. PMID- 11499954 TI - Inactivation of infectious pathogens in labile blood components: meeting the challenge. AB - Substantial improvement in the safety of blood transfusion has been achieved through the addition of new tests, such as nucleic acid tests, yet residual risk associated with transfusion of blood components persists. Transfusion of blood components has been implicated in the transmission of viruses, bacteria, and protozoa. While it is commonly recognized that hepatitis B virus (HBV), hepatitis C virus (HCV), cytomegalovirus (CMV), and the retroviruses, such as human immunodeficiency virus (HIV) and the human lymphotrophic viruses (HTLV) can be transmitted through cellular components, other pathogens are emerging as potentially significant transfusion-associated infectious agents. For example, transmission of protozoan infections due to trypanosomes and babesia have been reported. In addition to viral and protozoal infectious agents, bacterial contamination of platelet and red cell concentrates continues to be reported; and may be an under-reported transfusion complication. More importantly, new infectious agents may periodically enter the donor population before they can be definitively identified and tested for to maintain consistent safety of the blood supply. The paradigm for this possibility is the HIV pandemic, which erupted in 1979. During the past decade a number of methods to inactivate infectious pathogens in labile blood components have been developed and have entered the advanced clinical trial phase. PMID- 11499955 TI - Results of unrelated umbilical cord blood hematopoietic stem cell transplant. AB - The number of umbilical cord blood transplants (UCBT) is increasing worldwide, and the purpose of Eurocord is to evaluate the results and compare the outcome of UCBT with allogeneic bone marrow transplants (BMT). Data have been reported to Eurocord by many transplant centers. Close links have been established with cord blood banks through Netcord. BMT data have been provided by transplant centers and also by the European Blood and Marrow Transplant (EBMT) and International Bone Marrow Transplant Registries (IBMTR). Eurocord has analyzed the outcome of unrelated UCBT from 121 transplant centers and 29 countries. The results have shown that survival with unrelated mismatched UCBT was comparable to that with unrelated BMT. Engraftment with cord blood was delayed, resulting in an increased incidence of early transplant complications. The incidence of acute and chronic graft-versus-host-disease (GVHD) was reduced with cord blood grafts even in HLA mismatched transplants and in adults. In patients with leukemia, the rate of relapse was similar to that after BMT. The overall event-free survival with UCBT was not statistically different when compared to BMT. In conclusion, this large registry study confirms the potential benefit of using umbilical cord blood hematopoietic stem cells for allogeneic transplants. PMID- 11499956 TI - Platelet glycoproteins and their role in diseases. AB - The role of platelet glycoprotein receptors in disorders caused by their absence or defects such as in Bernard-Soulier syndrome or Glanzmann's thrombasthenia has been known for many decades now. Their function as targets for pathological antibodies is also well established. The possible roles of platelet receptors or their polymorphism variants in the origins of diseases such as cardiovascular disorders are less well studied. Investigation of this area began about five years ago and many findings still remain controversial. The involvement of platelet receptors in other diseases like asthma, diabetes and HIV are only starting to be studied. PMID- 11499957 TI - Structural and functional diversity of blood group antigens. AB - Biochemical and molecular genetic studies have revealed that blood group antigens are present on cell surface molecules of wide structural diversity, including carbohydrate epitopes on glycoproteins and/or glycolipids, and peptide antigens on proteins inserted within the membrane via single or multi-pass transmembrane domains, or via glycosylphosphatidylinositol linkages. These studies have also shown that some blood group antigens are carried by complexes consisting of several membrane components which may be lacking or severely deficient in rare blood group 'null' phenotypes. In addition, although all blood group antigens are serologically detectable on red blood cells (RBCs), most of them are also expressed in non-erythroid tissues, raising further questions on their physiological function under normal and pathological conditions. In addition to their structural diversity, blood group antigens also possess wide functional diversity, and can be schematically subdivided into five classes: i) transporters and channels; ii) receptors for ligands, viruses, bacteria and parasites; iii) adhesion molecules; iv) enzymes; and v) structural proteins. The purpose of this review is to summarize recent findings on these molecules, and in particular to illustrate the existing structure-function relationships. PMID- 11499958 TI - Insights into the epidemiology, natural history and pathogenesis of hepatitis C virus infection from studies of infected donors and blood product recipients. AB - Studies of patients with post-transfusion non-A, non-B hepatitis and their implicated donors were critical to the discovery of hepatitis C virus (HCV) and to the development and progressive enhancement of HCV immunoassays for donor screening and diagnostic applications. Post-transfusion HCV cases have also been used to define the time course of viral and immunological markers following acute infection. Even more precise data on the timing and characteristics of viremia and immune responses during primary HCV infection have been derived from studies of serial samples from source plasma donors who were identified while in the process of infection and seroconversion. Such data have been critical to the derivation of estimates for the viremic, pre-seroconversion window period, and hence projections of the yield of addition of nucleic acid amplification technology (NAT) or HCV core antigen enzyme immunoassays (Ag-EIAs) to antibody screening in donor and diagnostic settings. In addition to these implications for screening and diagnosis, studies of HCV in the blood donor and blood product recipient settings have yielded substantial insights into the epidemiology, pathogenesis and prognosis of HCV infection. In the future, prospective studies of blood and plasma donors detected in the primary phase of HCV infection by NAT screening, will be important for defining viral and host genetic and immunological determinants of clearance of acute viremia, as well as for investigating the benefits of early treatment. Thus, the findings from studies of HCV among donors and blood product recipients have yielded and should continue to provide important insights into HCV pathogenesis leading to novel diagnostic, therapeutic and vaccination strategies. PMID- 11499959 TI - Red blood cell transfusion strategies. AB - Although the hemoglobin level of 100 g/L has been used for many years as the allogeneic red blood cell (RBC) transfusion trigger, current evidence indicates that for most patients a more restrictive transfusion strategy is at least as effective as and possibly superior to a liberal transfusion strategy. Moreover, the available data indicate that the use of smaller volumes of allogeneic RBCs may be associated with decreased risk of morbidity and mortality. Thus several recent studies indicate that the use of more restrictive triggers than 100 g/L does not appear to adversely affect patient outcomes. Indeed, the majority of recently published RBC transfusion guidelines recommend a more conservative and cautious approach to allogeneic RBC transfusion practice, primarily to reduce the risk of transfusion-related adverse effects. However, the available transfusion trigger studies do not provide sufficient data to allow the claim that the improved outcomes observed are the sole result of the transfusion strategy used. It is possible that the results are the consequence of effects yet to be defined clearly. Additional studies will be necessary to determine the effects of RBC storage time and the presence of allogeneic leukocytes in allogeneic RBC transfusion practice. Nonetheless, the available data, together with detailed information about alternatives to blood product transfusions, will enable physicians to improve outcomes in transfused patients. PMID- 11499960 TI - Orthopaedic Surgery Transfusion Haemoglobin European Overview: the OSTEO study (extended abstract). PMID- 11499961 TI - Risk management: an important tool for improving quality. AB - Risk management implies that one has identified and analysed the root cause of the risk. In blood transfusion public and political opinion on the perceived risks are mainly related to product stigmatisation, to cognitive aspects of risk. Therefore this perception is affective, and has a negative connotation. In order to manage risk in an optimal manner, we need to understand how people think about it, and recognise that thoughts, feelings and behaviour are determined not only by psychological factors, but also by social, cultural and political influences. Perception of risk is always situated within a context, which may differ. Therefore people (i.e., public and political opinion) seem to act inconsistently from one risk context to another. Crucial for understanding the logic behind different risk perceptions is how people think about a hazard and organise information about it. The blood supply system has aspects that make it very vulnerable to crises of confidence, as the subject of blood can easily become stigmatised. The impact of the latter on the perception of blood transfusions and their recipients as well as the willingness of the public to accepttransfusions can be dramatic. Risk perception needs to be monitored in order to anticipate and adequately deal with public and political acceptance. We know that risk and stigmatisation are closely interconnected, and that the costs are likely to be high both for human health and for the maintenance of the healthcare system. Thus there is a global need to carefully monitor the safety of the blood supply systems and communicate risk information in a way that both informs people and builds up public and political confidence. It is therefore not sufficient to simply state that the blood supply is safe; it must also be made safe. So risk management becomes an integral part of quality management, as it deals with the public perception of the blood supply system and its respective elements: procurement and use. PMID- 11499962 TI - Quality and risk management: the commitment of the Council of Europe. PMID- 11499963 TI - Emerging viruses and transfusion. AB - The development of new technologies leads to the discovery of new viruses. For each of these new infectious agents, their possible relevance to blood transfusion needs to be assessed. The questions to be answered are transmissibility by transfusion, pathogenicity, prevalence in blood donors, persistence, and the availability of screening assays. Since 1995, three new viruses have been identified and extensively studied. PMID- 11499964 TI - The genetic variability of HIV-1 and its implications. PMID- 11499965 TI - Molecular diversity in the biosynthesis of GI tract glycoconjugates. A blood group-related chart of microorganism receptors. AB - This paper examines the potential of carbohydrate blood-group antigens present on mucosal surfaces in acting as receptors for microorganisms. Mucosal surfaces express significant amounts of carbohydrate blood-group antigens under the control of the Secretor, Lewis and ABO systems. The exact glycoconjugate profile an individual presents to the lumen is complex, and can only be correctly determined by a combination of serology and genotyping. We have isolated and structurally resolved the glycolipids expressed in the small intestine of group O individuals having various common or rare phenotypes. Using this information, we have been able to construct a biosynthetic pathway and propose that the type, size and glycotopes expressed, are controlled to a major extent by blood-group related glycosyltransferases. Many of these glycotopes are potential receptors for microorganisms; some resemble tumour antigens, while others resemble the lipopolysaccharides of some pathogens. Although the origins of the blood-group glycosyltransferases remain uncertain, it is evident that they significantly diversify the mucosal glycotopes exposed to microbes; and therein may be found a potential explanation for their existence. PMID- 11499966 TI - Non-myeloablative hematopoietic stem cell transplantation. AB - Conventional approaches to allogeneic stem cell transplantation have used toxic high-dose conditioning therapy in attempts to eradicate underlying diseases and achieve allogeneic engraftment. Preclinical studies and clinical observations have shown that to achieve engraftment conditioning regimens could be markedly reduced in intensity with reduction in treatment toxicities. The use of potent pre- and postgrafting immunosuppression facilitated stable mixed hematopoietic chimerism in a preclinical canine model. The initial clinical experiences with attenuated conditioning regimens have shown promise as a modality to achieve human stem cell transplantation with an improved safety profile. This may allow offering potentially curative hematopoietic stem cell transplantation (HSCT) to a more representative patient population (older and sicker) who are currently not eligible for such therapy. Obtaining a state of mixed hematopoietic chimerism could prove curative of the disease phenotype of various nonmalignant disturbances of the hematopoietic and immune systems. On the other hand, patients with hematopoietic malignancy will likely require conversion to full donor hematopoeisis by virtue of graft-versus-host (GVH) reactions directed against both recipient hematopoiesis and underlying malignancy. The infusion of additional donor lymphocytes has been proposed by many groups to augment graft versus tumor responses, but most likely more specific strategies will need to be developed to improve efficacy and avoid nonspecific GVH reactions. PMID- 11499967 TI - The impact of improved safety on maintaining a sufficient blood supply. AB - The impact of improved safety on maintaining a sufficient blood supply is becoming an increasingly real issue facing the National Blood Service in England and North Wales. This paper shows the extent of the impact that safety measures can have on reducing collection levels, without making any value judgement on the safety critera themselves. It demonstrates that underlying trends in collection are making it increasingly difficult to meet demand. Further potential restrictions, perhaps associated with variant Creutzfeldt-Jacob disease (vCJD), will require a reassessment of the safety-sufficiency trade-off. The analysis strongly suggests that continual enhancement of safety criteria, for both individual donor and patient benefit, will lead to insufficient supply to meet current levels of hospital demand. New approaches are needed to effectively manage blood throughout the supply chain, thereby ensuring sufficient supply. PMID- 11499968 TI - Nucleic acid testing: the international status of NAT. PMID- 11499970 TI - Evidence-based transfusion. PMID- 11499969 TI - Molecular nature of granulocyte antigens. AB - Granulocyte (neutrophil) antibodies can cause febrile and pulmonary (TRALI) transfusion reactions as well as immune neutropenias. In the last decade enormous efforts have been made to characterize the implicated alloantigens. The NA1, NA2, and SH antigens could be identified as polymorphic forms of the neutrophil Fc gammaReceptor IIIb encoded by three alleles. The antigens MART and OND have been located on the leucocyte adhesion molecules (beta2 integrins) and found to be caused by single nucleotide mutations in the alphaM (CD11b) and alphaL (CD11a) subunits. We have succeeded in throwing light on the primary structure of the NB1 antigen, which has recently been clustered as CD177. Based on these findings, the Granulocyte Antigen Working Party of the ISBT introduced in 1998 a new nomenclature for human neutrophil alloantigens (HNA nomenclature) based on the antigen's glycoprotein location. Elucidation of the molecular nature of granulocyte antigens now allows antigen identification by glycoprotein-specific immunoassays and genotyping by DNA techniques. Thus, considerable progress has been made in the characterization of granulocyte antigens. Further studies will improve our diagnostic tools and facilitate the prevention and management of transfusion reactions and immune neutropenias. PMID- 11499971 TI - Platelet transfusion trigger in difficult patients. AB - There is general consensus that a prophylactic pre-transfusion trigger at 10.000 platelets/microL in stable oncohematological patients is as safe as the traditional trigger of 20.000/microL, and that perioperative triggers at 50.000 and 100.000/microL are adequate in most surgical and neurosurgical conditions respectively. Guidelines on the trigger and other issues related to platelet transfusion can be found in nine documents published during 1987-2001 by the National Institutes of Health (NIH), the British Committee on Standardization in Hematology, the Royal College of Physicians of Edinburgh, the College of American Pathologists, the American Society of Anesthesiology and the American Society of Clinical Oncology (ASCO). Although consensus may be less evident on specific triggers for 'difficult' patients, the following triggers, listed by progressively increasing levels, have been proposed in the literature and have found general agreement: a stable oncohematological recipient: 10.000; lumbar puncture in a stable pediatric leukemic patient: 10.000; thrombocytopenia secondary to gpIIb/IIIa receptor inhibitors [corrected]:10.000; bone marrow aspiration and biopsy: 20.000; gastrointestinal endoscopy in cancer: 20.000 40.000; disseminated intravascular coagulation: 20.000-50.000; fiber-optic bronchoscopy in a bone marrow transplant recipient: 20.000-50.000; neonatal alloimmune thrombocytopenia: 30.000; major surgery in leukemia: 50.000; thrombocytopenia secondary to massive transfusion: 50.000; invasive procedures in cirrhosis: 50.000; cardiopulmonary bypass: 50.000-60.000; liver biopsy: 50.000 100.000; a nonbleeding premature infant: 60.000; neurosurgery: 100.000. The proposed values must be considered within the context of careful clinical evaluation of each individual patient, and attention should be given to the power of discrimination of platelet counters at low counts and to the prompt availability of good quality platelet products in the case of emergency. PMID- 11499972 TI - The therapeutic use of antibodies for malignancy. AB - The idea of using the specificity of antibodies to target malignant cells was put forward very soon after the discovery of techniques to generate monoclonal reagents. The responses seen with mouse anti-idiotype in patients with B-cell lymphomas indicated the potential of this approach, but it was some years before key technical obstacles were overcome and the more widespread application of these therapies became possible. Whilst they were originally conceived as having an immunotherapeutic effect, it has become clear that recruitment of immune effectors is only one component of successful antibody therapy, and their action upon the cellular target, either blocking or agonistic, is also critical. The development of immunoconjugates to deliver toxins or radiation is a further extension of the approach, and here again the intracellular effect of antibody ligation appears to be crucial. This presentation will address the central theme of antibody treatments for malignancy that are now reaching the clinic, and will use these examples to highlight ways in which antibodies may be acting in vivo. PMID- 11499973 TI - The safety of human blood: experimental TSE/prion infectivity studies. PMID- 11499974 TI - Platelet polymorphisms in thrombotic disorders. AB - Plaque rupture and/or endothelial damage lead to exposure of von Willebrand factor (VWF) and collagen which facilitate the adhesion of circulating platelets via glycoprotein (GP) GPIb-IX-V and integrin alpha2 beta1, respectively, to the damaged vessel wall. This process activates the platelets and leads to a conformational change of a second integrin alphaIIb beta3 that facilitates fibrinogen binding and platelet aggregation. Thrombin generated at the blood plaque interface converts fibrinogen to fibrin, which stabilizes thrombus growth. Therefore, any genetic differences that might alter surface expression or activity of these receptors could influence the risk for adverse outcome as a result of the hemostatic process. In the last five years, there has been a rapid accumulation of literature concerning the relationship between genetic variations in platelet glycoproteins and risk for coronary heart disease. In this study, we have presented a comprehensive review of the impact of platelet receptor polymorphisms and thrombotic risk. PMID- 11499975 TI - Novel platelet products, substitutes and alternatives. AB - Despite the many advances in the safety, processing, and storage of conventional 22 degrees C liquid-stored allogeneic platelet concentrates, there still are significant drawbacks to the use of such products. Efforts to overcome these shortcomings have resulted in an array of novel platelet products, substitutes, and alternatives; which are currently at various stages of development. This review summarizes the recent developments in the frozen and cold storage of platelets; their pathogen inactivation; as well as the status of lyophilized platelets, infusible platelet membranes (IPMs), red cells bearing arginine glycine-aspartic acid (RGD) ligands, fibrinogen-coated albumin microcapsules, and liposome-based agents; as potential alternatives to the use of conventional platelet transfusions. Pre-clinical studies have been encouraging for several of these novel products; however, to date, very few have entered human trials. Nonetheless, with the ongoing development of diverse products, those properties that may be necessary for their hemostatic effectiveness will become apparent. However, safety and efficacy must be demonstrated in pre-clinical and phase I to III clinical trials before these novel agents, substitutes and alternatives can be used clinically for patients with thrombocytopenia. PMID- 11499976 TI - Transfusion and lung injury. AB - The respiratory tree has been viewed as an infrequent site of injury arising as a complication of transfusion. In recent years, this view has changed as investigators have shown that two complications--circulatory overload and transfusion-related acute lung injury--are relatively frequent events. Circulatory overload is a result of hypertransfusion to individuals at risk, the very young or old recipient. The reaction is due to fluid infusion which overwhelms the capacity of the left ventricle, resulting in pulmonary edema. While rarely fatal, studies have shown that such incidents result in intensive care and extended hospitalization. In the setting of orthopedic surgery, 1% of elderly patients undergoing hip or knee surgery experience circulatory overload. These events are associated with autologous, as well as allogeneic red blood cells (RBC) and fresh frozen plasma. Transfusionists need to be vigilant with transfusion therapy in this population. Phlebotomy and supplemental oxygen are the key therapies. Transfusion-related acute lung injury (TRALI) is the adult respiratory distress syndrome due to transfusion. It is associated with a significant morbidity and mortality of 5-14%, making it the third most common cause of death from transfusion in developed countries. It is characterized by the onset of acute respiratory distress, bilateral pulmonary edema and hypoxemia. It occurs within 1-2 hours of transfusion of a plasma-containing blood product. All blood components have been associated with the reaction, and rarely, intravenous immune globulin. There is no recognized profile of individuals at increased risk for TRALI. There are two purported mechanisms of injury; the vast majority of cases are associated with passively transfused complement-activating antibodies. These antibodies are either HLA (Class I or II) or granulocyte specific. These antibodies appear to act as mediators, which result in granulocyte aggregation, activation, and microvascular pulmonary injury. With appropriate respiratory intervention, 80% of patients recover within 96 hours of the original insult. There are no permanent pulmonary sequelae. PMID- 11499977 TI - Mechanisms of severe transfusion reactions. AB - Serious adverse effects of transfusion may be immunologically or non immunologically mediated. Currently, bacterial contamination of blood products, particularly platelets, is one of the most significant causes of transfusion related morbidity and mortality. Septic transfusion reactions can present with clinical symptoms similar to immune-mediated hemolytic transfusion reactions and transfusion-related acute lung injury. Extremely high fever and/or gastrointestinal symptoms, in a transfusion recipient, may be indicative of sepsis. The diagnosis is based upon culturing the same organism from both the patient and the transfused blood component. Numerous organisms have been implicated as the cause of septic transfusion reactions. Due to different storage conditions, gram negative organisms are more often isolated from red blood cell components; gram positive organisms are more often isolated from platelets. Prevention of septic transfusion reactions is primarily dependent on an adequate donor history and meticulous preparation of the donor phlebotomy site. Visual inspection of blood components prior to transfusion is also vital to preventing these reactions. Several methods of detection of bacterial contamination and inactivation of pathogens are currently under active investigation. PMID- 11499978 TI - Yield and future issues of nucleic acid testing. AB - Despite the much lower actual yield than that estimated for hepatitis C virus (HCV) and human immunodeficiency virus (HIV) nucleic acid testing (NAT)-only positives in the USA and Germany, look-back procedures have revealed that no HCV transmission has occurred in Germany since the introduction of NAT. This indicates sufficient sensitivity of the pool-PCR approach. The slow ramp-up of hepatitis B virus (HBV) however, may require a different approach. It has been shown in Germany that the pooling of samples followed by virus enrichment results in a significant yield. Single donation testing for HBV would not increase the yield, because virus enrichment from mini-pool results in a similar sensitivity to that of single donation testing. Both strategies may be useful for extending future NAT to HBV screening. New candidate viruses for NAT are Parvo B19 and hepatitis A virus (HAV) because of their extreme resistance to inactivation procedures. Their low pathogenicity and epidemiologic characteristics, however, make them candidate viruses only for pooled source plasma. The main future issues of NAT will be related to the automation of pooling, extraction and amplification as a single homogeneous process. Depending on the throughput, automated single donation NAT as demonstrated by the 'Tigris' system may be an option, as far as all transfusion-relevant viruses will be included. In the near future high throughput systems will rely on pooled donor samples, most probably in conjunction with efficient enrichment procedures. For these systems, automation of the extraction and amplification process will be one of the first steps. These procedures will also limitthe costs of NAT and keep it available for use with future candidate viruses. PMID- 11499979 TI - Haemovigilance in Europe: the European Haemovigilance Network. PMID- 11499980 TI - DNA chips: a new tool for genetic analysis and diagnostics. AB - DNA chips are miniaturized microsystems based on the ability of DNA to spontaneously find and bind its complementary sequence in a highly specific and reversible manner, known as hybridization. Labeled DNA molecules in a sample are analyzed by DNA probes tethered at distinct sites on a solid support. The composition of the DNA sample is then deduced by analyzing the signal generated by labels present at each probe site. Applications are widespread: fundamental research, cancer or microbiology diagnostics, genotyping, gene expression, pharmacogenomics, and environmental control. Medical application consists, for example, in the identification and detection of mutations in genes responsible for cancers, or DNA chip analysis of individual polymorphisms which may provide a guide towards the most efficient treatment. In the environmental and agro industrial fields, DNA chips show great promise in rapidly testing microorganism content, contamination or pathogenicity. DNA chip dimensions offer hybridization sites in the 50-200 micron range, producing arrays ranging from 100 to 1,000,000 different probes per cm2. PMID- 11499981 TI - Universal leukoreduction of cellular and plasma components: process control and performance of the leukoreduction process. AB - Many countries in Europe and over the world are currently or will be concerned in the near future, by the implementation of universal leukoreduction (ULR) for red blood cells (RBC), platelets (PT) and now also for plasma. Recommended by several advisory committees, this decision to implement ULR must be considered as a recognition of the benefit of early leukocyte removal, and also as a precautionary measure to increase blood safety. The leukodepletion technology for RBC, PT and plasma has become increasingly more elaborated and integrated in the collection or in the component preparation process. To reach this aim and to assure that the end-products meet local specifications (1 or 5 x 10(6) residual leukocytes), a process control and validation program for leukoreduction has been described in the specific guidelines. Tested on a wide scale by a group of centers, flow cytometry is emerging as reference method for residual leukocyte enumeration. Validation protocols (linearity, precision, accuracy) have been defined in numerous national or international studies (PSL and BEST Working Party). The sensitivity of the method is greatly improved by concentration of the sample, with a detection limit equivalent to 10 cells/mL for RBC or PT, and 0.5 cells/mL for plasma. Furthermore, monitoring of the performance of the leukoreduction process includes a quality control program based on a general statistical model with a parametric or non parametric approach, sampling plan, ongoing control, process capability assessment, confidence limit, detection of failure, and estimation of the non conforming units rate. PMID- 11499982 TI - The future of plasma derivatives. AB - In the early 1980s, the advent of recombinant DNA technology provided the perspective of an unlimited supply of recombinant plasma proteins. However, the cost of mammalian cell expression, downstream processing, and the development time of biotechnological pharmaceuticals have generally been underestimated. Despite the initial optimism that plasma derivatives would soon become obsolete, no more than three plasma proteins have been licensed for therapeutic use since that time. The current situation favours a reappraisal of plasma derivatives, which nowadays can meet safety standards similar to those of recombinant products. In terms of cost, human blood has the further advantage over biotechnological sources in that it allows the production of multiple pharmaceutical proteins from one single material. This provides a favourable starting point for exploring the therapeutic potential of the numerous plasma proteins that have remained unused so far. PMID- 11499983 TI - Fetal DNA in maternal plasma: application to non-invasive blood group genotyping of the fetus. AB - The non-invasive determination of fetal genetic characteristics, including blood group types, is a long-sought goal of modern genetics. Previous work on the use of fetal cells in maternal blood has been hampered by the rarity of such cells. The recent discovery of cell-tree fetal DNA in maternal blood has opened up new possibilities for non-invasive prenatal diagnosis. It is particularly useful that fetal DNA is present in relatively high concentrations in maternal plasma, making its robust detection possible using modern technology. Large-scale clinical trials and standardization of protocols still need to be carried out. However, there is optimism that the accurate and safe prenatal determination of fetal blood group types may be achieved in routine clinical practice in the near future. PMID- 11499984 TI - Immune thrombocytopenia in the foetus and the newborn: diagnosis and therapy. AB - Thrombocytopenia is a common condition in intensive care units. However the frequency of neonatal thrombocytopenia in all newborns (< 150 x 10(9)/L) has been estimated at 1-4%. Foetal/neonatal immune thrombocytopenia due to the transplacental passage of maternal antiplatelet IgG is a transient passive disease in an otherwise healthy newborn. The major risk of severe thrombocytopenia is intracranial haemorrhage (ICH) leading to death or neurological impairment. The principal aim in the management of the affected infants is to prevent the deleterious consequences of severe thrombocytopenia. Autoimmune thrombocytopenic purpura (AITP) in pregnant women can induce moderate or severe thrombocytopenia in the foetus or the newborn whatever the mother's disease status. Foetal thrombocytopenia can occur as early as 20 weeks of gestation. The frequency of ICH has been estimated to be 1-3% of cases. Foetal thrombocytopenia cannot be prevented. After birth, thrombocytopenia usually worsens during the first days of life. Postnatal management is usually ly of intravenous immunoglobulins. Neonatal alloimmune thrombocytopenia is considered to be the platelet counterpart of haemolytic disease of the newborn. Severe bleeding in the central nervous system and death (10% of cases) or neurological sequelae (20% of cases) may occur. The incidence of neonatal alloimmune thrombocytopenia has been estimated at 1 per 800-1000 live births. After birth, maternal platelet transfusion is the treatment of choice. Due to the high risk of recurrence of foetal thrombocytopenia in subsequent pregnancies, protocols for antenatal management including maternal therapy with intravenous immunoglobulins and/or corticosteroids, or in utero transfusion have been proposed. PMID- 11499985 TI - Modulation of the alloimmune response by blood transfusions. AB - Blood transfusions can induce both immune activation and immunosuppression. The former is expressed by the induction of HLA alloantibodies and T cell activation, while the latter is accompanied by enhanced graft survival in transfused versus non-transfused recipients. The immunological mechanism leading to downregulation of the alloimmune response has not yet been elucidated. Possible explanations include the induction of a Th2 response by non-professional antigen presentation by the transfused blood cells and blockage of alloreactive T cell reactivity by soluble HLA and soluble FasL in the supernatant of blood components. These mechanisms, however, do not explain the observations which have shown that the degree of HLA compatibility between the transfusion donor and patient is a determining factor. Transfusions in which the donor blood shares at least one HLA DR antigen with the recipient induce tolerance, while fully HLA-DR mismatched transfusions lead to immunization. The importance of HLA-DR sharing suggests a central role for CD4+ regulatory T cells. In this case, indirect recognition of an allopeptide in the context of self-HLA-DR on the transfusion donor by CD4+ T cells of the recipient might be the clue to the induction of tolerance. Recent data from our laboratory in fact show that CD4+ T cells specific for an allopeptide in the context of self HLA-DR are able to downregulate the alloimmune response of autologous T cells. As these regulatory T cells produce IL-10, they may also be involved in the extension of tolerance via their modulatory effect on dendritic cells. It remains to be established whether these regulatory T cells are indeed responsible for the 'blood transfusion effect' in organ transplantation. PMID- 11499986 TI - Allorecognition. AB - Until recently, the vigorous T cell response via the direct pathway has overshadowed studies involving the indirect pathway. Thus, while the direct pathway has previously been considered to be the main driving force in alloimmune responses, there is an increasing body of data to support a prominent role of the indirect pathway in transplant rejection. Most importantly, while the direct anti donor alloresponse diminishes with time after transplantation, the indirect alloresponse is likely to be permanently active due to traffic of recipient dendritic cells (DCs) through the graft. Consequently, the future challenge in the induction of allograft tolerance is to design interventions that will target the cells involved in the indirect pathway, especially the T cells with indirect allospecificity. PMID- 11499987 TI - Effects of left frontal lesions on the selection of context-appropriate meanings. AB - The present study examined the role of the left frontal cortex in strategic aspects of semantic processing. Participants were tested in a semantic priming task involving the meaning access of ambiguous and unambiguous words. Patients with left or bilateral frontal lesions failed to develop semantic facilitation of context-appropriate homograph meanings relative to age-matched controls and patients with right frontal lesions who produced more facilitation than controls. When the ambiguous words, however, were replaced by unambiguous words, patients with left frontal lesions improved to normal levels of semantic priming. This pattern of results seems difficult to explain in terms of a problem to access semantic information per se or to use contextual cues. The findings are, however, consistent with a deficit in selecting context-appropriate meanings in the presence of competing meanings. PMID- 11499988 TI - Sex-related differences in event-related potentials, face recognition, and facial affect processing in prepubertal children. AB - Thirty-five prepubertal children, 17 boys and 18 girls, between the ages of 8 and 11 years, were studied to examine electrophysiological and cognitive sex differences during a face-recognition-memory (FRM) task and a facial-affect identification task (FAIT). All participants were prepubertal, as determined by J. M. Tanner's (1962) staging and endocrine evaluation. Sex-dependent event related potential (ERP) amplitude asymmetries were found during FRM. Boys displayed greater right versus left ERP amplitude to auditory tone probes during the task, whereas girls displayed the opposite pattern. In addition, positive correlations were obtained between ERP amplitude during FRM and FAIT accuracy scores for boys, but not for girls. Results suggest that girls and boys may use different neuronal systems in the processing of faces and facial affect. Findings are consistent with developmental theories regarding sex differences in visuospatial processing. PMID- 11499989 TI - Motor sequencing deficits in schizophrenia: a comparison with Parkinson's disease. AB - Motor abnormalities occur in schizophrenia (SZ) and may arise from striatal dysfunction. This study examined whether the pattern of performance on simple and complex motor abilities in SZ was similar to that of patients with Parkinson's disease (PD). Quantitative tests of speeded movement and motor and cognitive sequencing were used to assess 25 SZ, 16 PD, and 84 normal controls (NCs). Sequencing performance was also examined with motor rigidity taken into account. Compared with the NC group, the SZ and PD groups were impaired on measures of motor rigidity and motor sequencing. With rigidity accounted for, the SZ group was significantly more impaired than the PD group on motor sequencing; cognitive and motor processes contributed to the motor deficit. Cognitive sequencing performance predicted motor sequencing performance in PD but not SZ. Although both SZ and PD resulted in significant motor and cognitive sequencing deficits, the pattern and correlates of these deficits differ, suggesting that the affected neural systems underlying motor deficits in SZ are different from those involved in PD. PMID- 11499990 TI - Time perception and reproduction in young adults with attention deficit hyperactivity disorder. AB - Adults with attention deficit hyperactivity disorder (ADHD; n = 104) were compared with a control group (n = 64) on time estimation and reproduction tasks. Results were unaffected by ADHD subtype or gender. The ADHD group provided larger time estimations than the control group, particularly at long intervals. This became nonsignificant after controlling for IQ. The ADHD group made shorter reproductions than did the control group (15- and 60-s intervals) and greater reproduction errors (12-, 45-, 60-s durations). These differences remained after controlling for IQ and comorbid oppositional defiant disorder, depression, and anxiety. Only the level of anxiety contributed to errors (at 12-s duration) beyond the level of ADHD. Results extended findings on time perception in ADHD children to adults and ruled out comorbidity as the basis of the errors. PMID- 11499991 TI - Attentional asymmetry in schizophrenia: disengagement and inhibition of return deficits. AB - This research examined 2 components of visual orienting in medicated schizophrenia patients: the validity effect and the inhibition of return (IOR). In the 1st experiment, patients showed the expected asymmetry in orienting attention, that is, larger validity effect in the right visual field than in the left. However, this asymmetry was due to a deficit in facilitatory processes rather than a disengagement deficit. In addition, patients showed a deficit in IOR. In the 2nd experiment, a 2nd central cue for summoning attention, explicitly, back to the center was used. In this experiment, normal IOR in schizophrenia patients was found. Because it was shown that schizophrenia patients do not have a disengagement deficit, IOR possibly could not be observed because of the increased facilitation in that location. It was proposed that the abnormality in visual attention in schizophrenia is due to a deficit in inhibitory processes. PMID- 11499992 TI - The spectrum of category effects in object and action knowledge in dementia of the Alzheimer's type. AB - The validity and origin of category effects in the anomia demonstrated by individuals with dementia of the Alzheimer's type (DAT) remains controversial. Twenty DAT subjects were tested with picture naming and semantic association judgment tests. Picture and word stimuli were drawn from biological, nonbiological, and actions-verbs categories, all of equal difficulty and previously normed on elderly controls. DAT subjects made significantly more naming and semantic judgment errors in the biological category than in the nonbiological category. They were relatively more accurate in naming and making judgments for actions-verbs when presented as words or as 5-s animations. When line drawings of actions were shown for naming, performance deteriorated significantly. Converging results from these 2 tasks provide strong evidence for a semantic memory impairment preferentially affecting biological items to a greater extent than nonbiological items or action verbs in DAT. PMID- 11499993 TI - Are hemispheric strategy differences independent of the level of performance? AB - Studies requiring identification of consonant-vowel-consonant (CVC) nonwords have suggested that attention is distributed more slowly or sequentially across the letters when they are presented to the right hemisphere than to the left hemisphere. Two experiments investigated whether hemispheric differences in processing strategy would be reduced with reductions of hemispheric differences in accuracy. The magnitude of visual field differences in accuracy was controlled by manipulating exposure duration, and the effect was observed on visual field differences in processing strategy. For both CVC identification (Experiment 1) and identification of nonletter symbols (Experiment 2), hemispheric strategy differences were independent of differences in accuracy. Both quantitative and qualitative hemispheric differences in processing visual displays appear to depend on the nature of the stimuli and the nature of the processes they invoke. PMID- 11499994 TI - Emotion recognition from faces and prosody following temporal lobectomy. AB - The anteromedial temporal lobe has been found to participate in processing emotion, but there are unresolved discrepancies in the literature. To address this issue, the authors investigated recognition of emotion from faces and from prosody in 26 participants with unilateral temporal lobectomy (15 left, 11 right) and in 50 brain-damaged controls. Participants with right, but not left, temporal lobectomy did significantly worse in recognizing fear from facial expressions. There were no group differences in recognizing emotional prosody. Neither IQ nor basic perceptual function accounted for task performance; however, there was a moderate negative correlation between extent of amygdala damage and overall performance. Consistent with some prior studies, these findings support a role for the right anteromedial temporal lobe (including amygdala) in recognizing emotion from faces but caution in drawing conclusions from small group samples. PMID- 11499995 TI - Determinants of Benton Facial Recognition Test performance in normal adults. AB - Clinical investigations have found that lesions of the right cerebral hemisphere can disrupt face perception. Much less is known about the determinants of facial discrimination in healthy adults, although age-related differences in many cognitive abilities have been shown to correlate with simple processing speed and variation in regional brain volumes. In this study, 174 healthy adults between the ages of 20 and 92 were asked to match pictures of unfamiliar faces. After their performance was regressed on age, sex, education, and perceptual comparison speed, adding terms for frontal lobe volume, nonfrontal volume, and ventricle-to brain ratio (VBR) derived from magnetic resonance imaging improved the model and accounted for 35% of the variance in facial discrimination. VBR and processing speed alone accounted for nearly 34% of the variance. These findings suggest that both normal atrophic brain changes and decreases in processing speed contribute to individual differences in facial discrimination. PMID- 11499996 TI - Neonatal hypoxic risk in preterm birth infants: the influence of sex and severity of respiratory distress on cognitive recovery. AB - The authors examined the differential effects of neonatal respiratory distress syndrome (RDS)--a condition associated with elevated risk for neonatal hypoxia ischemia--on the cognitive development of the two sexes. The authors also attempted to establish whether the severity of respiratory distress is linked to cognitive outcome. Findings revealed an appreciable female advantage in cognitive recovery from RDS, yet the association between severity of RDS and outcome did not reach conventional statistical significance level. The sex effect on intelligence test performance was significantly greater for nonverbal subtests than for verbal ones. These results augment earlier findings of sex differences in cognitive recovery from perinatal intracranial hemorrhage. The current investigation also extends these results to a population of children at neonatal hypoxic risk for whom early brain injury was excluded on the basis of neonatal cranial ultrasound findings. PMID- 11499997 TI - Trichloroethylene in urine as biological exposure index. AB - Occupational exposure to trichloroethylene (TRI) was studied by analysis of environmental air and urine from 49 workers operating in a special printing house on glass. For the measurement of environmental concentration of TRI (Cenv), the ambient air was sampled using personal passive dosimeters. The activated charcoal was desorbed with carbon disulfide and injected into a gas-cromatograph - mass spectrometer (GC-MSD). The biological monitoring of exposed workers was conducted by determining the concentration of TRI in urine (Curine) Urine concentration of TRI was determined by headspace analysis using GC-MSD. Significant correlation was found between the environmental TRI concentration and urinary TRI concentration. The use of a regression equation between Curine (microg/l) and Cenv (mg/m3) (Curine = 0.081 x Cenv + 4.27) resulted in a value of Curine corresponding to Threshold Limit Value-Time Weighted Average (TLV-TWA) exposure value (269 mg/m3) of 26.0 microg/L. PMID- 11499998 TI - Toluene alters mu-opioid receptor expression in the rat brainstem. AB - Toluene is an ototoxic organic solvent widely used in industry and could be a cause of sleep apnea. Acute toluene administration in rats induces an increase in the number of neural cells immunostained for mu-opioid receptors in several brainstem nuclei, such as the inferior colliculus, dorsal and lateral periaqueductal gray and dorsal raphe, without changes in the superior colliculus and the interpeduncular and lateral reticular nuclei. These data suggest that mu opioid receptors could be involved in toluene-induced neurotoxic effects on the physiological regulation of breathing during sleep, and auditive function. PMID- 11499999 TI - Induction of gonadal toxicity to female rats after chronic exposure to mancozeb. AB - Mancozeb, a fungicide of ethylenebisdithiocarbamate group was orally administered at doses of 500, 600, 700 and 800 mg/kg body weight/day to normal virgin rats of Wistar strain for 30 days. The vaginal smear and body weight of the rats were recorded daily and rats were sacrificed on 31st day. Estrous cycle was effected by showing a significant decrease in the number of estrous cycle, duration of proestrus, estrus and metestrus with concomitant significant increase in the duration of diestrus in all the mancozeb treated groups when compared with controls. There were a significant decrease in the number of healthy follicles and a significant increase in the number of atretic follicles in all the mancozeb treated groups when compared with controls. The histologic observation of the ovary revealed the presence of less number of corpora lutea and the size of the ovary was also reduced in high doses of mancozeb treated rats. There was a significant increase in the thyroid weight in all the mancozeb treated rats except in 500 mg/kg/d. In rats treated with 500 mg/kg/d showed a significant increase in the level of total lipids in the liver. In rats treated with 600 mg/kg/d mancozeb showed a significant decrease in the levels of glycogen and total lipids in the uterus and total lipids in the liver. In rats treated with 700 mg/kg/d showed a significant decrease in the levels of protein in ovary, glycogen, total lipids, phospholipids and neutral lipids in the uterus and a significant increase in the levels of phospholipids, neutral lipids in the ovary and total lipids, phospholipids and neutral lipids in the liver. In rats treated with 800 mg/kg/d showed a significant decrease in the levels of protein and glycogen in the ovary and protein, glycogen, total lipids, phospholipids and neutral lipids in the uterus and a significant increase in the levels of total lipids, phospholipids and neutral lipids in the ovary and liver when compared with controls. These observed effect of mancozeb on the estrous cycle, follicles and biochemical constituents may be due to imbalance in the hormone or toxic effect. PMID- 11500000 TI - Regular overtime and cardiovascular functions. AB - It was concluded that there was sufficient evidence about a possible link between long working hours particularly exceeding 50 a week and the risk of significant health outcomes, including cardiovascular disease from literature review by Spurgeon et al.. This study was conducted to find out the single effect of regular overtime work on the cardiovascular functions through objective biological indices such as blood pressure or heart rate variability. We conducted a field survey of 238 male engineers who were working at the department of research & development of three electronics manufacturing companies in S. Korea. The field survey consisted of (1) self-report questionnaire (working hours and health conditions, and fatigue) and (2) measurements of blood pressure and heart rate variability. By multivariate analyses we could show the relationship between overtime work and some cardiovascular functions after controlling the effects of major confounders such as age and sleeping hours, which were pointed out by Iwasaki et al. (1998) and Sasaki et al. (1999). Especially, low frequency component (power in the low frequency range, 0.04-0.15 Hz) of the heart rate variability during work might be used as early objective biological indices for chronic effect of regular overtime work on cardiovascular functions. However, we should confirm those effect through the well-designed prospective study. PMID- 11500001 TI - Long working hours and subjective fatigue symptoms. AB - Presuming that overtime work may associate with chronic fatigue and then decrease cardiovascular function, this report was prepared to find out the definite positive relationship between long working hours and subjective fatigue complaints. We analysed the data of a field survey of 238 men who were working at the department of research & development of three electronics manufacturing companies in S. Korea. The field survey consisted of self-report questionnaires on the working hours, health conditions, and fatigue. For data analysis, the subjects were divided into 3 groups on the criteria of 60 and 70 working hours per week: less longer (LLWH), longer (LWH), and more longer (MLWH) working hour groups. We compared the age-adjusted mean % scores of fatigue complaints among LLWH, LWH, and MLWH. The rate of complaints of subjective fatigue before going to work for LWH and MLWH tended to be significantly higher than those for LLWH. Hence, we conclude that the questionnaire on the subjective fatigue complaints is a good screening tool for early detection of cumulative fatigue due to chronic job stress such as long working hours. PMID- 11500002 TI - A comparison of vibration magnitudes on the tool with different subject according to the ISO 8662-7 standard. AB - Recently, ISO has defined the guidelines for the evaluation of the measurement and exposure limits of hand-transmitted vibrations at the handle of hand-held portable power tools. In a measurement of vibrations at pneumatic impact wrenches that tighten and remove screws. ISO 8862-7 standard: 1997 and ISO 5349 standard: 1986 are set as international standards. In order to make an exact measurement of the quantities of vibrations exposed to the hand-arm system of a human body, ISO 8662-7 gives specifications to a measurement device and a loading device. As to a measurement device, for instance, the mass, installation method etc. of transducers and mechanical filters are defined. In addition, it specifies with each tolerance the working conditions of a pneumatic tool, the number of revolutions of socket in a loading device, the feed force, the number of subjects, and such an evaluation method as how to decide a declaration value of vibrations. These specifications give toolmakers a comparative measurement of vibration values of their tools before shipment. Meanwhile, although the measurement condition described in ISO8662-7: 1997 is that skilled operators conduct vibration measurement, no provisions as to operators are made in detail. Hence, this study focuses its concern on ambiguous work experience years in measuring vibrations of the tools in conformity with ISO 8662-7, and hereby the effect of years of work experience upon measurement values is considered and at the same time a knowledge of how to select operators is made clear. PMID- 11500003 TI - Occupational hypersensitivity pneumonitis due to isocyanates: mechanisms of action and case reports in Japan. AB - Isocyanates are very useful chemicals, and these are important for our daily life. Various products, especially urethane resin, are made from these chemicals. There is no substitute for isocyanates at the present. Isocyanates are patent agents to provoke immunological responses. This article deals with the case reports of hypersensitivity pneumonitis (HP) induced by isocyanates in Japan. It is said that HP is one of the rare diseases induced by isocyanate with very low frequency so far in the world. Certainly there are rare cases of isocyanate-HP also in Japan, but these are the cases only in large enterprises that pay attention to their working places. In Japan, there are several cases of isocyanate-induced HP, which happened in minor enterprises such as those with less than 50 workers. In clinical appearance, there are few conflicts with Western studies. The best choice for sensitized people is isolation from working places. It is important for the workers to understand the diseases. In these days, several tests for diagnosis are developed. It is indispensable to put these tests for prevention of these diseases and improvement of their working places. PMID- 11500004 TI - A new approach to assess low frequency noise in the working environment. AB - To assess high-level low frequency noise in the working environment, adverse extra-aural effects caused by the noise should be taken into account. The human body vibration induced by low frequency noise, 'noise-induced vibration', was measured on the body surface and the equal-acceleration level contours of the vibration were tentatively estimated. With these contours, we can predict the magnitude of noise-induced vibration at every measuring position on the body surface. This is helpful in relating the total dosage of low frequency noise with the physical symptoms caused by the noise. But some important points in the contours remain to be investigated and improved. When these points are dealt with, the equal-acceleration level contours will be useful for assessing high level low frequency noise in the working environment from the standpoint of predicting the adverse extra-aural effects. PMID- 11500005 TI - National occupational health research strategies. Report of "the Conference on Occupational Health Research Strategies in the 21st Century", supported by Ministry of Health, Labour and Welfare. PMID- 11500006 TI - Aging: fact or fiction? PMID- 11500007 TI - Management of hearing loss in infants: the UTD/Callier Center position statement. AB - As infants are being identified earlier and earlier by universal newborn hearing screening programs, there is a need to establish principles for professionals involved in habilitative processes. Recently, a panel of experts was convened for a 2-day conference entitled "Habilitative Issues for Infants with Hearing Loss," the Bruton Conference on Audiology/Communication Sciences, UTD/Callier Center for Communication Disorders. The purpose of this document was to summarize the panel's discussions on habilitative principles, state-of-the-art practices, and future needs for infants with hearing loss. PMID- 11500008 TI - Hearing sensitivity in adults screened for selected risk factors. AB - As a means of partially distinguishing age effects and other risk factors in presbyacusis, hearing thresholds for 56- to 65-year-old participants screened for exclusion of selected risk factors were compared with thresholds for 48- to 55 year-old participants without the same risk factors. Hearing thresholds for both age groups were also compared with age-appropriate International Standards Organization (ISO) norms. Even after screening participants for exclusion of selected risk factors, differences in hearing sensitivity remained across age groups. Across the male and female groups, thresholds were generally better (lower) for 48 to 55 year olds than for 56 to 65 year olds. ISO norms generally underestimated the degree of hearing loss for participants of comparable age in the present study. The underestimation was small at lower test frequencies, was generally greater for men, and was largest at test frequencies above 2000 Hz. PMID- 11500009 TI - Possible neuronal refractory or recovery artifacts associated with recording the mismatch negativity response. AB - The mismatch negativity (MMN) is an event-related potential that involves a negative voltage shift of baseline electroencephalographic (EEG) activity in the approximate latency window of the N1 and P2 cortical potentials in response to new or novel sounds. The MMN is present at birth and has been hypothesized to serve as an automatic preconscious detector of changes in the auditory environment. Research paradigms used to extract the MMN response from EEG activity have a potential problem related to neuronal refractoriness or recovery. Both N1 and P2 are known to increase in amplitude with longer interstimulus intervals (ISIs). The MMN extraction procedures involve mathematical subtraction of waveforms elicited by standard sounds (with short ISIs) from those recorded to rare deviant sounds (with longer ISIs). Any ISI-dependent amplitude changes in N1 and/or P2 could therefore alter the morphology of the resulting difference wave and lead to misinterpretation of the nature of the underlying MMN generators. We tested 12 young females and found that the MMN can be influenced by ISI-dependent refractory effects that may modify the waveform morphology. This has important clinical implications since the MMN is being investigated as an assessment tool. PMID- 11500010 TI - Evaluation of a cognitive test battery in young and elderly normal-hearing and hearing-impaired persons. AB - A cognitive test battery sensitive to processes important for speech understanding was developed and investigated. Test stimuli are presented as text or in an auditory or audiovisual modality. The tests investigate phonologic processing and verbal information processing. Four subject groups, young/elderly with normal-hearing and young/elderly with hearing impairment, each including 12 subjects, participated in the study. The only significant effect in the text modality was an age effect in the speed of performance, seen also in the auditory and audiovisual modalities. In the auditory and audiovisual modalities, the effects of hearing status and modality were seen in accuracy parameters. Interactions between hearing status and modality, both in accuracy and in reaction times, show that hearing-impaired subjects have difficulties without visual cues. Performing the test battery in noise made the tasks more difficult, especially in the auditory modality and for the elderly, affecting both accuracy and speed. Test-retest measurements showed learning effects and a modality dependent variability. The test battery has proven useful in assessing the relative contribution of different input signals and the effects of age, hearing impairment, and visual contribution on functions important for speech processing. PMID- 11500011 TI - Effects of speech babble on transient evoked otoacoustic emissions in normal hearing adults. AB - The results of transient evoked otoacoustic emissions (TEOAEs) have been found to be adversely affected by noise. However, there are few investigations into the specific effect of background noise on TEOAEs. The purpose of the present study was to determine the effects of increasing levels of speech babble and the recording method on TEOAEs using the ILO88 Otodynamics instrumentation. Subjects were 30 normal-hearing adults (15 males and 15 females), aged between 18 and 32 years. TEOAE recordings were obtained from both ears of each subject under five different levels of speech babble delivered via a loudspeaker in sound field using both the default and Quickscreen methods of data collection. The results indicated that both the whole-wave reproducibility (WWR) and mean signal-to-noise ratio (MSNR), averaged across frequencies from about 2 to 4 kHz, decreased with increasing speech babble levels. The results also showed that the Quickscreen mode was less susceptible to noise than the default mode. The pass percentage, using either WWR > or = 50 percent or an MSNR > or = 3 dB as a pass criterion, also decreased with increasing speech babble levels. Furthermore, the findings indicated that the use of the WWR pass criterion resulted in a high failure rate under high levels of speech babble. In contrast, the MSNR pass criterion was robust to speech babble levels of up to 70 dBA in the Quickscreen mode and 65 dBA in the default setting. The clinical implications of these findings, as applied to non-sound-treated environments, are discussed. PMID- 11500012 TI - Tinnitus handicap inventory. PMID- 11500013 TI - Effect of arsenite on certain aspects of protein metabolism in fresh water teleost, Tilapia mossambica (Peters). AB - The sublethal toxicity of sodium arsenite on protein metabolism was investigated in teleost fish, Tilalpia mossambica at the end of 24, 48, 72 and 96 h of exposure. Total protein content, free amino acid content and activities of the enzymes aspartate amino transferase (AAT) and alanine amino transferase (ALAT) in liver, gill, brain and muscle exhibited significant (P<0.05) alterations throughout the investigation in relation to that of control. It is suggested that the fish is able to respond to the stressful situations by gearing up the metabolic activity as revealed by the elevated protein, amino acid content and the activities of AAT and ALAT. PMID- 11500014 TI - Toxic effects of copper and zinc mixtures on some haematological and biochemical parameters in common carp, Cyprinus carpio (Linn). AB - The effect of heavy metals and mixture were studied on some haematological and biochemical parameters in fish, Cyprinus carpio at sublethal level over the period of 30 days. Heavy metal significantly decreased total RBC count, haemogoblin, heamatocrit (Hct) (except copper after 10 days exposure). The WBC count was increased significantly in all the treated fish. The MCH, MCHC, MVC were increased depending upon the exposure period; declined PVC was noticed at 1% level of significance after 30 days on all the treated fish. Serum level of chloride and calcium content were increased where as serum glucose, cholesterol and total serum protein were decreased significantly. The alteration in haematological and biochemical parameters shows more toxic in mixture after 30 days, showed more than 'addition action' of the individual toxicant. PMID- 11500015 TI - Influence of animal manures on the biology of temperate earthworm, Eisenia fetida in tropical semiarid climate. AB - For understanding the potential utility in field scale production of vermicompost and vermiprotein economically, Eisenia fetida was cultured to establish the influence of (i) prevailing tropical semiarid (North-East region of Karnataka, India) environmental factors and (ii) different animal manures (cattle, horse and 1:1 mixture of cattle and horse) (on its growth, reproduction and life span. In three forms of diet, growth (mg/d/g live weight of worm) was almost similar, but the biomass in cattle manure (565.7 +/- 15.3) was significantly more than horse manure (494.9 +/- 22.8) and 1:1 mixture (470.3 +/- 22.0). Mean cocoon production (per worm/week) in horse manure (0.16) was significantly (P<0.001) lower than that in cattle manure (1.6) and in 1:1 mixed manure (1.4). Cattle favoured biomass, growth and cocoon production. Horse manure inhibited cocoon production and the conserved energy in this process might have been added to the body weight almost equal to that in cattle manure. In 1:1 mixed manure, the biomass was less due to moderate cocoon production (presumably due to the stimulatory influence of 50% cattle manure) under semifavorable nutrients and environmental conditions. Fecundity of the worms declined with aging, despite favourable nutrients and environmental conditions. Worms survived up to 92, 68 and 66 weeks in cattle manure, 1:1 mixed manure and horse manure respectively. PMID- 11500016 TI - Bioconversion of 1,3 dinitrobenzene by Micrococcus colpogenes strain MCM B410: quantitation and characterization of the intermediates. AB - Micrococcus colpogenes MCM B 410, indigenous to soil, collected from nitro aromatic contaminated site, could transform 1,3 dinitrobenzene (m-DNB) initially to m-nitroaniline, m-nitrophenol, m-aminophenol and resorcinol at 30 degrees C under shake culture condition. Carbon mineralization studies with unlabelled and radio labelled 1,3 (U14 C) dinitrobenzene subtrates indicated that the above metabolities appeared within 4 days. After 7 days incubation a significant traction of the source compound was degraded to C Q through aliphatic acids. Presence of nitro aryl reductase, aryl monooxygenase and resorcinol 1,3 di oxygenase was also noted in the sonicated cell mass. PMID- 11500017 TI - Health problems among workers of iron welding machines: an effect of electromagnetic fields. AB - The possible effects of EMFs on 100 workers were studied by means of structured interview and rating of subjective symptoms. As control 41 sewing machine operators and assembly workers were chosen, interviewed and likewise tested. The present Indian ceiling value of 250 Tesla for the equivalent power density was exceeded in more than 50% of the machines. The highest leakage fields, for EMFs, were found near machines, which gave a high exposure to the hands. Eye irritation complaints were reported by 40% of the workers. The fertility outcome did not show any significant result. PMID- 11500018 TI - Effect of sewage water on the growth and yield parameters of wheat and blackgram with different fertilizer levels. AB - Impact of biologically treated domestic sewage water was studied on the growth and yield attributing characters of wheat and blackgram under different fertilizer levels in the field condition. There was no significant difference on the yield and yield attributing characters of wheat. However, yield attributing characters like number of tillers/plant, panicle length (cm), number of grains/panicle and 1,000 grain weight showed increasing trend to the tune of 13.29, 1.59. 5.49 and 3.79 per cent respectively over the tubewell irrigated crop. Yield was also increased to the tune of 3.63 per cent when the crop was irrigated with sewage water. Interactions between irrigation and fertilizer levels were also insignificant. However, sewage water with N60P45K45 responded better on tillering, panicle length, number of grains/panicle and yield of wheat. In case of blackgram, treated sewage water had no significant effect on the grwoth and yield attributing characters, rather sewage water had some negative effect on these parameters. Interactions between irrigation and fertilizer levels were insignificant. But tubewell irrigation with N10P10K10 was more responsive on growth parameters. When the crop was grown with tubewell irrigation and without fertilizer, yield attributing characters were more positively responded. PMID- 11500019 TI - Effect of rubber factory effluent on seed germination and seedling growth of Vigna radiatus L. AB - Physico-chemical analysis of the rubber factory effluents revealed high amounts of total suspended and dissolved solids. Sulphate, phosphate, total nitrogen were also present in significant amounts. At higher concentration (above 50%) of effluent, the seed germination percentage was retarded. Diluted effluent (upto 50%) favoured seedling growth. Length of root system, shoot system and number of lateral roots were increased by low concentrations of effluent. PMID- 11500020 TI - Effects of some trace heavy metals on Poecilia reticulata (Peters). AB - Three trace heavy metals viz., nickel, copper and zinc were studied for their toxic action against Poecilia retriculata (Peters). Among these, copper was found to be most active followed by zinc and nickel. Accumulations of these metals as well as behavioural studies were carried out after exposing fish to sub-lethal concentration of LC20. It was found that the highest quantity of nickel was accumulated in the fish body followed by zinc and copper. After exposure to sub lethal concentration of metals some behavioural changes in fish were observed due to stress, such as mucus like secretion over gills, excessive excretion, anoretic condition and increased distance between gills and operculum. In all the cases fin movement was observed. Role and use of such changes as biological indicators or as biological early warning system in water quality assessment has been discussed. PMID- 11500021 TI - Metalaxyl effect on nitrogenase activity (acetylene reduction) and yield of mungbean (Vigna radiata (L.) wilzek). AB - In an experiment, application of different levels of metalaxyl to a sandy loam soil significantly affected the nodulation and nitrogenase activity of mungbean. In both the compost amended and unamended soils, 0.5 mg kg(-1) of metalaxyl enhanced acetylene reduction activity and yield of mungbean, where as higher concentrations (1 mg and 2.5 mg kg(-1) of fungicide) inhibited the nodulation traits as well as economic traits of mungbean. PMID- 11500022 TI - Environmental effect on the reoccurrence of Alternaria alternata on Trianthema portulacastrum. AB - Trianthema portulacastrum is a noxious weed which spreads rapidly in the field. Pathogen Alternaria alternata was observed on this host for three years continuously i.e. 1997 to 1999. The incidence of the pathogen depended on rainfall and temperature. PMID- 11500023 TI - Gill lesions in the perch, Anabas testudineus, exposed to monocrotophos. AB - Histopathological effects of sublethal doses of monocrotophos on the gills have been studied by exposing the fish for a period ranging from ten to twenty days. The extent of damage to gills was dependent on the dose and duration of exposure. Histopathological changes in the gills observed were characterized primarily by hemorrhage in the primary and secondary gill lemellae. Degeneration and necrosis of epithelial cells were very prominent. Distortion of the secondary lamellae was very prominent, beginning with disruption of epithelial cells from pillar cells. PMID- 11500024 TI - Breeding biology of Megalops cyprinoides from Visakhapatnam coast. AB - Studies on stages of maturity, gonado-somatic indices and breeding cycle from two different habitats viz., the polluted Harbour Waters and the unpolluted Yerragedda Mouth revealed that M. cyprinoides breeds twice a year i.e., during Jan-Jul and Dec-Jan. Size at first maturity was 18 cm. in females and 19 cm. in males at both the stations. Sex ratio was 1:1 at the polluted station and 1:1.1 at the unpolluted station indicating a slight domination of females. PMID- 11500025 TI - The role of tissue transglutaminase in the germinal vesicle breakdown of mouse oocytes. AB - We have investigated the novel function of tissue transglutaminase (tTG) in the germinal vesicle breakdown (GVBD) of mouse oocyte. tTG was identified in ooplasm and germinal vesicle by immunostaining with less amount in germinal vesicle. Spontaneous maturation of the oocytes elevated in situ activity of tTG by over 2.5-fold at 3 h, which was determined by a confocal microscopic assay. However, incubation with monodansylcadaverine (MDC), a tTG inhibitor, blocked the activation of tTG. The possible role of tTG in GVBD was investigated by the use of two tTG inhibitors, MDC and cystamine. MDC largely inhibited the GVBD by a concentration-dependent manner. GV-stage oocytes were matured to the GVBD stage by 78% at 3 h in the normal culture condition. However, in the oocytes incubated with MDC for 3 h, the GVBD rates were 43 and 11% by 50 and 100 microM, respectively. MDC also blocked the entry of 70 kDa RITC-dextran from the ooplasm to the compartment of germinal vesicle, indicating a possible inhibition of nuclear pore disassembly by MDC. The role of tTG in GVBD was further investigated by microinjection with cystamine. The control oocytes, injected with DPBS, showed about 80% of GVBD at 3 h. But the oocytes injected with cystamine showed 15% of GVBD at 3 h and a little higher rate at 6 h. In addition, the inhibition of GVBD maturation by MDC was reversible by washing. These results suggested that tTG was involved in the early event of mouse oocyte maturation. PMID- 11500026 TI - The cyclooxygenase hydroperoxide product PGG(2) activates synaptic nitric oxide synthase: a possible antioxidant response to membrane lipid peroxidation. AB - Nitric oxide is a potent inhibitor of membrane lipid peroxidation. It is unknown, however, whether nitric oxide synthase (NOS) activity increases under conditions of membrane lipid peroxidation. Importantly, cyclooxygenase (COX)-catalyzed peroxidation of arachidonic acid is well-established to be increased by lipid hydroperoxides. The results of the present study demonstrate that the COX hydroperoxide product prostaglandin G(2) (PGG(2)) greatly stimulated NOS activity in synaptosomal membrane fractions from rat brain in a dose-dependent (EC(50) = 0.2 microM) manner in the presence of ATP and the antioxidant urate. NOS activation was also produced, albeit to a lesser extent, by 15 hydroperoxyeicosatetraenoic acid (15-HPETE) but not by the corresponding hydroxy compounds PGH(2) and 15-HETE or by hydrogen peroxide. These findings demonstrate that PGG(2)-activated synaptic NOS by a hydroperoxide-mediated pathway and support the view that NOS activation may be an important physiological response to lipid peroxidation. PMID- 11500027 TI - The effect of dexamethasone on P450 activities in regenerating rat liver. AB - The aim of our study was to detect four P450s (CYP1A, CYP2B, CYP2E1, CYP3A) on the basis of selective enzyme activities and protein amount, and to investigate the effect of dexamethasone treatment during liver regeneration. Partial hepatectomy of rats resulted in the loss of CYP1A, CYP2B, CYP2E1, and CYP3A activities. The reduction of enzyme activities and the loss of enzyme protein of CYP2B1/2, CYP2E1, and CYP3A1/2 were the most pronounced. In the case of CYP1A1, only slight decrease was observed. Dexamethasone treatment seems to counteract this loss mainly in the first 12 h. PMID- 11500028 TI - Sialylation of human IgG-Fc carbohydrate by transfected rat alpha2,6 sialyltransferase. AB - A recombinant IgG3 antibody with Phe-243 replaced by Ala (FA243) was expressed in a CHO-K1 parental cell line. The resulting IgG-Fc-linked carbohydrate was significantly alpha2,3-sialylated (53% of glycans), as indicated by normal- and reverse-phase HPLC analyses. Following transfection of a rat alpha2,6 sialyltransferase gene into this parental cell line, IgG-Fc-linked glycans were sialylated (60% of glycans) such that the ratio of alpha2,6- to alpha2,3-linked sialic acid was 0.9:1.0. By comparison, the wild-type IgG3 (F243) is minimally sialylated (2-3% alpha2,3-linked), thus suggesting that sialylation is controlled primarily by the protein structure local to the carbohydrate and that the two sialyltransferases compete to sialylate the nascent oligosaccharide. The additional alpha2,6-sialylation affected the function of the recombinant antibody. FA243 IgG3 having both alpha2,6 and alpha2,3-sialylation restored recognition to wild-type IgG3 levels for human FcgammaRI, FcgammaRII, and target cell lysis by complement. We discuss how sialylation linkage could modulate IgG function. PMID- 11500029 TI - Cloning, chromosomal mapping, and characteristic 5'-UTR sequence of murine cytosolic sialidase. AB - We have totally sequenced a cytosolic sialidase [EC 3.2.1.18] by RT-PCR from the murine thymus (murine thymic sialidase, MTS) which has a 1844-base length (encoding 385 amino acids including two sialidase motifs) and is the longest cytosolic sialidase ever reported. MTS has high and relatively low homologies with those of mammalian cytosolic sialidases from the mouse brain (99%), rat (91%), and human skeletal muscle (75%), and those of the mouse lysosomal (47%) and membrane-bound (51%) sialidases, respectively. Chromosomal mapping, being the first report of mouse cytosolic sialidase gene, showed that the MTS gene is localized to the distal part of mouse chromosome 1D and to rat chromosome 9q36. RT-PCR with the site-specific primers revealed that the coding region was expressed in all organs tested, but expressions including the 5'-UTR were barely detectable except for in the upper-thymic fraction. Also, soluble sialidase activity in the thymus was the highest of these organs. There were mRNA instability signals and AT-rich regions in 143 bp of MTS 5'-end. PMID- 11500030 TI - A novel bradykinin-related peptide from skin secretions of toad Bombina maxima and its precursor containing six identical copies of the final product. AB - Amphibian skin contains rich bradykinin-related peptides, but the mode of biosynthesis of these peptides is unknown. In the present study, a novel bradykinin-related peptide, termed bombinakinin M, was purified from skin secretions of the Chinese red belly toad Bombina maxima. Its primary sequence was established as DLPKINRKGPRPPGFSPFR that comprises bradykinin extended from its N terminus by a 10-residue segment DLPKINRKGP. The cDNA structure of bombinakinin M was found to contain a coding region of 624 nucleotides. The encoded precursor of bombinakinin M is composed of a signal peptide, an acidic peptide, six 100% identical copies of a 28-amino-acid peptide unit including bombinakinin M plus a spacer peptide. The sequence of bombinakinin M is preceded by a single basic residue (arginine), which represents the site of cleavage for releasing of mature bombinakinin M. This is the first cDNA cloning of bradykinin-related peptides from amphibian skin. The unique cDNA structure encoding bombinakinin M suggests that the generation modes of bradykinin-related peptides in amphibian skin and in mammalian blood system are different. PMID- 11500031 TI - Intercalating fluorescence dye YOYO-1 prevents the folding transition in giant duplex DNA. AB - Recently, it has become clear that with the addition of polyamines, giant DNA molecules of size greater than 10 kbp exhibit all-or-none switching between elongated coil and folded compact states. Here the effects of the intercalating fluorescent labeling dye, YOYO-1, and the minor-groove binding fluorescent labeling dye, DAPI, on the folding transition of single giant T4 DNA (166 kbp) induced by spermidine(3+) were examined, by use of the experimental technique of single molecular chain observation with fluorescence microscopy. It is found that the intercalating dye, YOYO-1, markedly prevents the folding transition, whereas the minor-groove binding dye, DAPI, exhibits negligible effect on the folding transition. This action of YOYO-1 is discussed in relation to the biological effect of intercalators. PMID- 11500032 TI - Del1: a new protein in the superficial layer of articular cartilage. AB - Articular cartilage contains four distinct zones, extending from the surface to the subchondral bone. Freshly isolated chondrocytes from the superficial zone of articular cartilage retain a collagenase-P-resistant cell-associated matrix. In the studies described here, the protein Del1 was identified as a component of the cell-associated matrix of superficial zone chondrocytes from adult bovine articular cartilage. Very little Del1 was associated with freshly isolated deep zone chondrocytes. Western blot analysis of articular cartilage cell and tissue extracts using polyclonal antibodies specific for Del1 showed Del1 was present in an insoluble cell-associated fraction. Extracts of the superficial zone of articular cartilage were found to be enriched in Del1 compared to the deeper layers of the tissue. Immunohistochemical staining of full-thickness articular cartilage with anti-Del1 antibodies also showed an enrichment of Del1 in the superficial zone. These observations are the first to describe the protein Del1 in a nonendothelial, nonfetal tissue. PMID- 11500033 TI - Downregulation of phospholipase C delta3 by cAMP and calcium. AB - Four different isoforms of mammalian phospholipase C delta (PLCdelta) have been described. PLCdelta1, the best-understood isoform, is activated by an atypical GTP-binding protein. It has been suggested that it is a calcium signal amplifier. However, very less is known about other subtypes, including PLCdelta3. Therefore, in the present study, we examined the expression of PLCdelta3 in different human tissues. Moreover, the cellular underlying regulation for PLCdelta3 was studied in different cell lines. Our study showed that the mRNA and protein levels differed significantly among human tissues. The human PLCdelta3 gene was composed of 15 exons and 1 putative cAMP response element in the 5'-end promoter region. PLCdelta3 mRNA expression was downregulated by cAMP and calcium in both the human normal embryonic lung tissue diploid WI38 cell line and the glioblastoma/astrocytoma U373 cell line. However, mRNA expression showed no impact by PKC activators or inhibitors. This study shows the human PLCdelta3 expression pattern and is the first report that PLCdelta3 gene expression is downregulation by cAMP and calcium. PMID- 11500034 TI - B lymphocytes and plasma cells express functional E-selectin by constitutive activation of NF-kappaB. AB - E-selectin (CD62E), a cell adhesion molecule for most leukocytes, is known to be expressed exclusively on the cytokine-stimulated endothelial cells mainly by inductive activation of NF-kappaB. Using immunohistochemistry and in situ hybridization, we showed that B lymphocytes and plasma cells in the spleens and lymph nodes from nude mice (T-lymphocyte-deficient), but not from SCID mice (T- and B-lymphocyte-deficient), expressed E-selectin prior to cytokine stimulation. The expression of E-selectin was also confirmed on human B lymphocytes isolated from peripheral bloods. The mouse J774A.1 monocytes could adhere to the marginal zones of mouse spleens in an E-selectin Ab inhibitable manner, suggesting the functional activity of the expressed E-selectin. In addition, ARH-77 cells, a cell line derived from human plasma cells, were found to express E-selectin mRNA and protein and to have a NF-kappaB activity for an E-selectin promoter. NF kappaB antagonists, such as TPCK (tosylsulfonyl phenylalanyl chloromethyl ketone), dexamethasone and a IkappaBalpha mutant plasmid could inhibit both the NF-kappaB activity and the expression of E-selectin. Transfection with an E selectin promoter-driven reporter gene construct further verified the E-selectin promoter activity in ARH-77 cells. Again, TPCK, dexamethasone, and the IkappaBalpha mutant plasmid could neutralize this activity. These findings suggest that B lymphocytes and plasma cells can express E-selectin, which is functional for monocytic leukocytes, by a mechanism of constitutive activation of NF-kappaB. PMID- 11500035 TI - Iba1 is an actin-cross-linking protein in macrophages/microglia. AB - Iba1 is a 17-kDa EF hand protein that is specifically expressed in macrophages/microglia and is upregulated during the activation of these cells. When exposed to macrophage colony-stimulating factor (M-CSF), microglia cell line MG5 immediately produces intense membrane ruffles in which Iba1 accumulates together with filamentous actin. In this study, we investigated the physical interaction between Iba1 and actin by centrifugation assay and electron microscopic examination and showed that Iba1 possesses actin-binding and -cross linking activities. Inhibitory mutant Iba1 that suppresses M-CSF-induced membrane ruffling had lost the actin-cross-linking activity, and it inhibited the cross linking activity of intact Iba1. These results indicate that Iba1 is a macrophage/microglia-specific actin-cross-linking protein essential for M-CSF induced membrane ruffling. PMID- 11500036 TI - Phosphorylation of serine-rich protein encoded by open reading frame 3 of the TT virus genome. AB - TT virus (TTV) is a newly discovered human virus with a single-stranded, circular DNA genome. The TTV DNA sequence includes two major open reading frames (ORFs), ORF1 and ORF2. Recently, spliced TTV mRNAs were detected and revealed two additional coding regions, ORF3 and ORF4. We found sequence similarity between the TTV ORF3 protein and hepatitis C virus (HCV) nonstructural 5A (NS5A) protein, which is a phosphoprotein and is thought to associate with various cellular proteins. To test whether the TTV ORF3 protein is phosphorylated, the state of phosphorylation was analyzed with a transient protein production system. The TTV ORF3 protein was phosphorylated at the serine residues in its C-terminal portion. Furthermore, the TTV ORF3 gene generated two forms of proteins with a different phosphorylation state, similar to the HCV NS5A region, suggesting that TTV ORF3 protein has function(s) similar to phosphorylated viral proteins such as the HCV NS5A protein. PMID- 11500037 TI - Cholesteryl ester transfer protein deficiency causes slow egg embryonation of Schistosoma japonicum. AB - In our attempt to discover a potential cause for accumulation of cholesteryl ester transfer protein (CETP) deficiency in Eastern Asia, we studied the association of CETP deficiency with pathogenesis of Schistosoma japonicum, a life threatening parasite peculiar to this region. The eggs of S. japonicum showed slow embryonation when cultured in CETP-deficient human plasma. Restoration of CETP to the deficient plasma rescued it, while inhibition of CETP in normal plasma did not cause slow embryonation of the cultured eggs. The egg embryonation was also retarded in the liver but not in the intestine of wild-type mice in comparison to the CETP-transgenic mice. The granulomatous lesion around the parasite eggs in the liver was less in the wild-type than in the CETP-transgenic mice. Thus, CETP deficiency may act against Schistosomiasis japonica by retarding egg embryonation, a potential cause of liver granulomatosis. It does not seem directly due to the lack of CETP activity in plasma but to abnormal lipoprotein generated by chronic CETP deficiency. PMID- 11500038 TI - Direct observation of release of cytochrome c from lipid-encapsulated protein by peroxide and superoxide: a possible mechanism for drug-induced apoptosis. AB - Release of cytochrome c from inside lipid vesicles and from inside proteoliposomes formed by cytochrome c oxidase has been studied by spectrophotometric methods. The protein encapsulated inside vesicles did not form complex with sodium azide solution added externally. Both hydrogen peroxide and superoxide were found to cause release of cytochrome c from the lipid encapsulated protein, which was detected from the distinct spectral changes due to the formation of the azide complex of cytochrome c in the solution. Cytochrome c encapsulated inside proteoliposomes containing cytochrome c oxidase (CcO) did not release the cytochrome c during enzymatic turnover of CcO. The anticancer drug, doxorubicin, was found to inhibit the biochemical function of cytochrome c oxidase and release of cytochrome c was observed from the proteoliposome encapsulating the protein during the enzymatic turnover in the presence of doxorubicin. The results indicated that the inhibition of enzymatic activity by doxorubicin possibly leads to the formation of reactive oxygen species, which induce the release of cytochrome c from inside to outside of the membrane. PMID- 11500039 TI - Spatial organization of Ca(2+) entry and exocytosis in mouse pancreatic beta cells. AB - Secretion from single pancreatic beta-cells was imaged using a novel technique in which Zn(2+), costored in secretory granules with insulin, was detected by confocal fluorescence microscopy as it was released from the cells. Using this technique, it was observed that secretion from beta-cells was limited to an active region that comprised approximately 50% of the cell perimeter. Using ratiometric imaging with indo-1, localized increases in intracellular Ca(2+) concentration ([Ca(2+)](i)) evoked by membrane depolarization were also observed. Using sequential measurements of secretion and [Ca(2+)](i) at single cells, colocalization of exocytotic release sites and Ca(2+) entry was observed when cells were stimulated by glucose or K(+). Treatment of cells with the Ca(2+) ionophore 4-Br-A23187 induced large Ca(2+) influx around the entire cell circumference. Despite the nonlocalized increase in [Ca(2+)](i), secretion evoked by 4-Br-A23187 was still localized to the same region as that evoked by secretagogues such as glucose. It is concluded that Ca(2+) channels activated by depolarization are localized to specific membrane domains where exocytotic release also occurs; however, localized secretion is not exclusively regulated by localized increases in [Ca(2+)](i), but instead involves spatial localization of other components of the exocytotic machinery. PMID- 11500040 TI - The N-terminal internal region of BLM is required for the formation of dots/rod like structures which are associated with SUMO-1. AB - Bloom Syndrome (BS) is a human autosomal genetic disorder characterized by a predisposition to a variety of malignant tumors. The gene responsible for BS encodes a protein (BLM) consisting of 1417 amino acids with a nuclear localization signal in the C-terminal region, which is a member of the RecQ helicase family. We previously showed, using a yeast two-hybrid system, that BLM interacted with Ubc9, which is the conjugating enzyme of SUMO-1 (small ubiquitin related modifier-1). In the present study, we exogenously expressed a green fluorescent protein-tagged Bloom syndrome protein, GFP-BLM, in human 293EBNA cells and found that it formed dots/rod-like structures associated with SUMO-1 in the nucleus. Deletion experiments indicated that the region from amino acids 238 to 586 of BLM is required for the formation of dots/rod-like structures associated with SUMO-1, and the DNA helicase domain, but not the helicase activity itself, slightly affected the formation and/or stability of these structures. Expression of a GFP-BLM which contained the 238-586 region, but lacked the C-terminal nuclear localization signal, resulted in localization to the cytoplasm without the formation of dots/rod-like structures and association with SUMO-1, indicating that these events occur only in the nucleus. PMID- 11500041 TI - Determinants of shear stress-stimulated endothelial nitric oxide production assessed in real-time by 4,5-diaminofluorescein fluorescence. AB - The extremely short biological half-life of endothelial-derived nitric oxide (NO) has impeded real-time measurements of NO synthesis. We used the membrane permeable fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA) to study determinants of NO synthesis in bovine aortic endothelial cells (BAECs). A step increase in shear stress (SS) from 0.3 to 3.4 dyne/cm(2) triggered an increase in DAF-2 fluorescence starting 3.0 +/- 0.5 min after the flow rise and peaking at 44.7 +/- 7.2 min. This was abolished by intracellular Ca(2+) chelation, but was unaffected by blocking extracellular Ca(2+) influx or by inhibiting SS-related changes in intracellular pH. The increase in DAF-2 fluorescence occurred significantly earlier in BAECs transfected with either superoxide dismutase (SOD) or catalase (CAT), indicating concomitant reactive oxygen species (ROS) generation by SS and "competition" between ROS- and DAF-2-NO interactions. These data provide novel insights into several NO signaling determinants and reveal that DAF-2 can assess real-time SS-stimulated NO synthesis in endothelial cells. This should facilitate the analysis of NO-signaling pathways. PMID- 11500042 TI - Increased retinoid signaling in vascular smooth muscle cells by proinflammatory cytokines. AB - Retinoids have been shown to modulate inflammation and the immune response in many cell types including macrophages, endothelial cells, and vascular smooth muscle cells. However, present knowledge of whether inflammatory mediators modulate vitamin A status in these cells is limited. To identify the role of inflammation on retinoid metabolism in vascular smooth muscle cells, the cells were exposed to a combination of proinflammatory cytokines: interleukin-1beta, interferon-gamma, and lipopolysaccharides. Without stimulation with proinflammatory cytokines, vascular smooth muscle cells expressed retinol dehydrogenases-2 and 5 mRNA detected by RT-PCR. Stimulation with the combination of cytokines induced a substantial increase of retinol dehydrogenase-5 mRNA. This was associated with increased production of ligands for retinoic acid receptors, when assayed in a retinoic acid receptor-dependent luciferase reporter system. Our results demonstrate that inflammatory mediators activate the retinoid metabolic pathway in vascular smooth muscle cells, which potentially may modulate the inflammatory response in the vascular wall. PMID- 11500043 TI - Recombinant expression, purification, and kinetic characterization of chondroitinase AC and chondroitinase B from Flavobacterium heparinum. AB - Glycosaminoglycans (GAGs) are a family of complex polysaccharides involved in a diversity of biological processes, ranging from cell signaling to blood coagulation. Chondroitin sulfate (CS) and dermatan sulfate (DS) comprise a biologically important subset of GAGs. Two of the important lyases that degrade CS/DS, chondroitinase AC (EC 4.2.2.5) and chondroitinase B (no EC number), have been isolated and cloned from Flavobacterium heparinum. In this study, we outline an improved methodology for the recombinant expression and purification of these chondroitinases, thus enabling the functional characterization of the recombinant form of the enzymes for the first time. Utilizing an N-terminal 6x histidine tag, the recombinant chondroitinases were produced by two unique expression systems, each of which can be purified to homogeneity in a single chromatographic step. The products of exhaustive digestion of chondroitin-4SO(4) and chondroitin-6SO(4) with chondroitinase AC and dermatan sulfate with chondroitinase B were analyzed by strong-anion exchange chromatography and a novel reverse-polarity capillary electrophoretic technique. In addition, the Michaelis-Menten parameters were determined for these enzymes. With chondroitin-4SO(4) as the substrate, the recombinantly expressed chondroitinase AC has a K(m) of 0.8 microM and a k(cat) of 234 s(-1). This is the first report of kinetic parameters for chondroitinase AC with this substrate. With chondroitin-6SO(4) as the substrate, the enzyme has a K(m) of 0.6 microM and a k(cat) of 480 s(-1). Recombinantly expressed chondroitinase B has a K(m) of 4.6 microM and a k(cat) of 190 s(-1) for dermatan sulfate as its substrate. Efficient recombinant expression of the chondroitinases will facilitate the structure-function characterization of these enzymes and allow for the development of the chondroitinases as enzymatic tools for the fine characterization and sequencing of CS/DS. PMID- 11500044 TI - Reaction of human myoglobin and peroxynitrite: characterizing biomarkers for myoglobin-derived oxidative stress. AB - Mixtures of human myoglobin (Mb) (or the Y103F variant of human Mb), authentic peroxynitrite (ONOO(-), ONOO(-):protein 2 mol/mol), and 5,5-dimethyl-1-pyrroline N-oxide (DMPO) gave radicals adducts at cysteine-110 (DMPO-C110) that are detected directly by electron paramagnetic magnetic spectroscopy (EPR). DMPO-C110 was detected exclusively over a range of DMPO concentrations (DMPO:protein ratios 25-100 mol/mol). Treatment of human Mb (or Y103F Mb) with the ONOO(-) generator 5 amino-3-(4-morpholinyl)-1,2,3-oxadiazolium (SIN-1) chloride (ONOO(-):protein 5 mol/mol) yielded a cross-linked Mb dimer as judged by SDS-PAGE analyses. Addition of DMPO or carbonate effectively eliminated the cross-linked product. Mass analyses of samples containing human Mb (or Y103F Mb), carbonate, and ONOO(-) indicated that nitration occurs exclusively at Y103. Thus, reaction of human Mb and ONOO(-) yields specific products that depend on the presence or absence of physiological concentrations of carbonate. These products may serve as biomarkers for the participation of Mb-derived radicals in the oxidative damage associated with myocardial reperfusion injury. PMID- 11500045 TI - Occurrence of a putative SCF ubiquitin ligase complex in Drosophila. AB - Many proteins are targeted to proteasome degradation by a family of E3 ubiquitin ligases, termed SCF complexes, that link substrate proteins to an E2 ubiquitin conjugating enzyme. SCFs are composed of three core proteins-Skp1, Cdc53/Cull, Rbx1/Hrt1-and a substrate specific F-box protein. We have identified in Drosophila melanogaster the closest homologues to the human components of the SCF(betaTrCP) complex and the E2 ubiquitin-conjugating enzyme UbcH5. We show that putative Drosophila SCF core subunits dSkpA and dRbx1 both interact directly with dCu11 and the F-box protein Slmb. We also describe the direct interaction of the UbcH5 related protein UbcD1 with dCul1 and Slmb. In addition, a functional complementation test performed on a Saccharomyces cerevisiae Hrt1p-deficient mutant showed that Drosophila Rbx1 is able to restore the yeast cells viability. Our results suggest that dRbx1, dSkpA, dCullin1, and Slimb proteins are components of a Drosophila SCF complex that functions in combination with the ubiquitin conjugating enzyme UbcD1. PMID- 11500046 TI - Light scatter and DNA accessibility to propidium iodide of ataxia telangiectasia and fanconi anemia cells. AB - Cells from individuals with genetic diseases ataxia telangiectasia (AT) and Fanconi anemia (FA) exhibit hypersensitivity to ionizing radiation (AT) or DNA cross-linking agents (FA) which may be caused by multiple factors including defects in chromatin structure and DNA repair. In this study, a combination of cytometric techniques was employed to study the chromatin conformation of AT and FA cells. Nuclei of peripheral blood mononuclear cells (PBMCs) and of skin fibroblasts established from AT and FA patients were analyzed by light scattering and fluorimetric titration with the DNA-intercalating dye propidium iodide. The light scatter measurements revealed the presence of small-sized nuclei with reduced granularity in PBMCs and fibroblasts from both AT and FA patients. The fluorometric titration data could be interpreted by assuming two classes of propidium iodide binding sites with different affinities. The number of high affinity sites in AT and FA fibroblasts was significantly larger (by 20%) than in control cells. Our findings show the applicability of cytometric techniques for the rapid assessment of chromatin conformation and also suggest the possibility to identify AT and FA carriers. PMID- 11500047 TI - Phorbol ester downregulates PDGFbeta receptor via PKCbeta1 in vascular smooth muscle cells. AB - The role of protein kinase C (PKC) and their isoforms in cell growth regulation remains elusive. Here we showed that in cultured human vascular smooth muscle cells (SMC), the PKC stimulator phorbol 12-myristate 13-acetate (PMA) inhibited [(3)H]thymidine incorporation in response to the growth factor PDGF associated with downregulation of PDGFbeta (but not alpha) receptors, which was recovered to normal level after PKC was depleted. The changes in PDGFbeta receptor were inversely correlated with PKCbeta1 protein levels regulated by PMA. The downregulation of PDGFbeta receptor by PMA was fully prevented by the PKCbeta inhibitor LY379196, however, without recovery of [(3)H]thymidine incorporation to PDGF. In contrast, [(3)H]thymidine incorporation was fully recovered after depletion of PKCs. These results indicate that in human SMC PKCbeta1 mediates PDGFbeta receptor downregulation. Other PKC isoforms activated by phorbol ester also contribute to the inhibitory effects on cell growth. PMID- 11500048 TI - Inactivating properties of recombinant ROMK2 channels expressed in mammalian cells. AB - Biophysical properties of ROMK2 channel were investigated at physiological temperature, after reexpression of the recombinant ROMK2 protein in a mammalian cell expression system (COS-7). We observed that ROMK2 induced an inwardly rectifying K(+) current whether polyvalent cations were present or not. Above +10 mV, ROMK2-induced current exhibited a voltage- and time-dependent decay, consistent with an inactivation process. Inactivation of ROMK2-induced current was also seen in inside out patch from ROMK2-expressing Xenopus oocyte. In COS-7 cells, inactivation was found to account for most of the inward rectification. Mg(2+) and spermine modulated rectification by accelerating inactivation kinetics independently of membrane potential. These results establish for the first time ROMK2 properties in a mammalian cell expression system. PMID- 11500049 TI - Identification and characterization of the promoter region of the GRM3 gene. AB - We have recently described the genomic organisation of the human metabotropic glutamate receptor 3 (GRM3) gene. The putative promoter region is characterised by the presence of a CCAAT and Sp1 site and the absence of a TATA box. Using a reporter gene assay, now we describe the functional activity of GRM3 promoter by transient transfection in both human neuroblastoma and astroglioma cell lines. Deletion of the CCAAT box and Sp1 site resulted in a pronounced reduction of reporter gene expression in both cell types, which indicates that these elements to correspond to the core promoter region. Moreover, we found that the genomic sequence 140 bp upstream of the first transcription initiation site appears to contain regulatory promoter elements for a preferential transcription of the gene in neuroblastoma cells. We also provide evidence that the genomic sequence spanning exon I, corresponding to the GRM3 5'-untranslated region, contains a negative regulatory element that represses gene transcription. PMID- 11500050 TI - ET(B) receptor activates adenylyl cyclase via a c-PLA(2)-dependent mechanism: a novel counterregulatory mechanism of ET-induced contraction in airway smooth muscle. AB - Endothelin-1 (ET-1) contracted the rabbit tracheal smooth muscle (RTSM), yielding a bell-shaped tension-concentration curve. Moreover, ET-1 induced concentration- and time-dependent increases in cAMP concentrations in RTSM (EC(50), 58 nM; t(1/2), 2.4 min). Pretreatment with the AC inhibitors, SQ-22536, or 2'-5' dideoxyadenosine, enhanced contraction to ET-1 and converted its bell-shaped tension curve into a sigmoidal one, but left contraction to carbachol and KCl unaltered. The potent ET(B)-receptor agonists, ET-3 or sarafotoxin-c, mimicked ET 1's effects on cAMP levels (EC(50) values 55 and 50 nM). Further, cAMP formation by ETs was inhibited by BQ-788 (selective ET(B) receptor blocker; IC(50), 8 nM), but not by BQ-610 (selective ET(A) receptor blocker). Removal of the epithelium did not prevent ET-induced increases in cAMP levels. Unlike isoproterenol, ETs failed to activate AC in membrane fractions from RTSM. In intact RTSM, the c PLA(2) inhibitor, AACOCF3, and the cyclooxygenase inhibitor, indomethacin, blocked ET-induced increases in cAMP levels. These findings reveal a novel, nonepithelial, c-PLA(2)-mediated, regulatory mechanism downstream from ET(B) receptors. PMID- 11500051 TI - Cloning and characterization of F-LANa, upregulated in human liver cancer. AB - Differentially expressed genes between normal liver and hepatocellular carcinomas were investigated using differential display. Consequently, we identified a fragment cDNA upregulated in tumor tissues. We screened the liver library and cloned the full-length cDNA, named F-LANa. Increased expression of F-LANa was confirmed by Northern blot analysis in 10 of 14 (71%) cases of hepatocellular carcinomas. Human F-LANa gene maps to chromosome 17p at D17S1828-D17S786, spans at least 11.8 kb, and contains 7 exons. This gene encodes a 239 aa protein exhibiting 97.9% similarity to the mouse ortholog gene, identified later by in silico cloning. Homology analysis was carried out in various species and showed that F-LANa was evolutionarily conserved from yeast to human. In addition, F-LANa antisense oligonucleotide suppressed F-LANa expression in human hepatocellular carcinoma BEL-7404 cells and significantly inhibited cell growth. Together, our data demonstrate that overexpression of evolutionarily conserved F-LANa occurs frequently and may play an important role in proliferation. PMID- 11500052 TI - Appearance of shortened Bcl-2 and Bax proteins and lack of evidence for apoptosis in rat forebrain after severe experimental traumatic brain injury. AB - To investigate whether apoptosis plays a role in traumatic brain injury (TBI), we examined the expression of Bcl-2 and Bax proteins and the release of mitochondrial cytochrome c in rat brains using Western blot analysis. Bcl-2 at the predicted 26 kDa was not detected in controls and TBI groups. However, at 1 h post-TBI, a shortened Bcl-2 protein with a molecular size of approximately 14.5 kDa was detected in the injured hemisphere (R). At 4 and 12 h post TBI, an additional bcl-2 band ( approximately 10 kDa) was detected in R. Both bands disappeared at 14 days post-injury. The predicted 21-kDa band of Bax was detected in both controls and TBI animals. In addition, two shortened Bax proteins ( approximately 18 kDa) were detected after TBI. The time course of appearance was similar to that of Bcl-2 described above. In the present study, neither cytochrome c release from mitochondria nor DNA fragmentation was detected in the forebrains of sham and TBI groups. Treatment of animals with an antioxidant N acetylcysteine administered ip greatly diminished the levels of shortened Bcl-2 and Bax proteins. These findings suggest that the induction of shortened Bcl-2 and Bax proteins in rat brains may be associated with reactive oxygen species generated after TBI. PMID- 11500053 TI - Reversal of cisplatin and multidrug resistance by ribozyme-mediated glutathione suppression. AB - gamma-Glutamylcysteine synthetase (gamma-GCS) is a key enzyme in glutathione (GSH) synthesis, and is thought to play a significant role in intracellular detoxification, especially of anticancer drugs. Increased levels of GSH are commonly found in the drug-resistant human cancer cells. We designed a hammerhead ribozyme against gamma-GCS mRNA (anti-gamma-GCS Rz), which specifically down regulated gamma-GCS gene expression in the HCT-8 human colon cancer cell line. The aim of this study was to reverse the cisplatin and multidrug resistance for anticancer drugs. The cisplatin-resistant HCT-8 cells (HCT-8DDP cells) overexpressed MRP and MDR1 genes, and showed resistance to not only cisplatin (CDDP), but also doxorubicin (DOX) and etoposide (VP-16). We transfected a vector expressing anti-gamma-GCS Rz into the HCT-8DDP cells (HCT-8DDP/Rz). The anti gamma-GCS Rz significantly suppressed MRP and MDR, and altered anticancer drug resistance. The HCT-8DDP/Rz cells were more sensitive to CDDP, DOX and VP-16 by 1.8-, 4.9-, and 1.5-fold, respectively, compared to HCT-8DDP cells. The anti gamma-GCS Rz significantly down-regulated gamma-GCS gene expression as well as MRP/MDR1 expression, and reversed resistance to CDDP, DOX and VP-16. These results suggested that gamma-GCS plays an important role in both cisplatin and multidrug resistance in human cancer cells. PMID- 11500054 TI - Escherichia coli CopA N-terminal Cys(X)(2)Cys motifs are not required for copper resistance or transport. AB - Escherichia coli CopA is a Cu(I)-translocating P-type ATPase that is involved in copper export and resistance. It is an orthologue of the human Menkes and Wilson disease-related proteins. Each of those two human copper pumps has six N-terminal Cys(X)(2)Cys sequences, but their function in transport is unclear. CopA has two N-terminal Cys(X)(2)Cys sequences, GLSC(14)GHC(17) and GMSC(110)ASC(113). The requirement of these cysteine motifs was investigated by mutagenesis of the codons for all four cysteine residues, singly and in combination. Cells of a copA deletion strain expressing genes for the mutant genes were nearly as resistant to copper as the wild type. In addition, everted membrane vesicles from cells expressing the mutant copA genes exhibited ATP-coupled accumulation of copper similar to that of the wild type. The results indicate that neither of two N terminal Cys(X)(2)Cys motifs is required for either resistance or transport. PMID- 11500055 TI - Chronic hypoxia induces apoptosis in cardiac myocytes: a possible role for Bcl-2 like proteins. AB - The effect of prolonged hypoxia as well as the molecular mechanisms on cardiac cell death is not well established. A possible role of Bcl-2 and Bax in hypoxia induced apoptosis in different cell types has been proposed. Here we demonstrate the effect of hypoxia on the induction of apoptosis and the expression of Bcl-2 like proteins in vivo and in vitro. Hearts from rats exposed to chronic hypoxia (n = 4) showed an increased rate of apoptosis compared to normoxic hearts (n = 4). The induction of apoptosis in hypoxic hearts correlated with a significant decrease of Bcl-2 protein level, whereas Bax protein expression was increased. Exposure of isolated neonatal rat cardiac myocytes to hypoxia also resulted in a significant increase in apoptosis. However, Bcl-2 and Bax protein levels essentially remained unchanged. Our results may suggest a different molecular mechanism of hypoxia-induced apoptosis in vivo and in vitro. PMID- 11500056 TI - Menin interacts directly with the homeobox-containing protein Pem. AB - The tumour suppressor gene causing multiple endocrine neoplasia type 1 (MEN1) encodes a 610 amino acid protein, menin. In order to identify menin-interacting proteins we used a yeast two-hybrid assay to screen a 12.5-dpc mouse embryo library with partial menin encompassing amino acids 278 to 476. This identified a homeobox containing protein encoded by a placenta and embryonic expression gene, referred to as Pem. GST-pull-down and coimmunoprecipitation experiments confirmed the interaction. Both proteins colocalised predominantly in the nucleus but were occasionally also found in the cytoplasm. Furthermore, in situ hybridisation studies revealed similarities in their expression patterns in mouse embryos and adult tissues. In adult mice both Men1 and Pem yielded strong signals in testis, Sertoli cells and particularly in seminiferous tubules. Thus, our study has identified that menin interacts with Pem, and the high expression of these proteins in the testis suggests a role in spermatogenesis. PMID- 11500057 TI - A novel system to identify Myb target promoters in friend murine erythroleukemia cells. AB - Friend murine erythroleukemia (MEL) cells provide an early erythroid precursor model that can be induced to terminally differentiate in cell culture and has been used to study erythroid differentiation as well as multistage tumorigenesis. During the chemically induced differentiation of MEL cells, expression of the c myb protooncogene is downregulated in a biphasic fashion and forced expression of c-myb is able to block the differentiation process, suggesting that c-myb activity may be limiting for differentiation in MEL cells. We have recently produced stable transfectants in the C19 MEL cell line that carry a dominant interfering myb allele (MEnT) under the control of an inducible mouse metallothionein I (MTH) promoter. Upon inducing expression of MEnT, transfected cells enter a differentiation program and begin to produce alpha-globin mRNA, assemble hemoglobin, and stop proliferating. Differential display was used to compare mRNA expression between parental C19 MEL cells induced to differentiate with hexamethylene bisacetamide (HMBA) and stable transfectants induced to differentiate via expression of MEnT to identify potential Myb target promoters. We identified six candidate cDNAs in this fashion and present evidence that two of these represent genes that are dependent on c-Myb activity for maximal expression in MEL cells. PMID- 11500058 TI - The role of transcriptional activation in the function of the Drosophila myb gene. AB - Vertebrate myb genes encode DNA-binding proteins that regulate transcription and have been implicated in regulation of cell proliferation, differentiation, and apoptosis. We have demonstrated that the single myb gene in Drosophila melanogaster, Dm myb, is required for the G(2)/M transition of the cell cycle and for suppression of endoreduplication. Recently, it has become apparent that the family of proteins containing Myb-related DNA-binding domains is much larger than originally believed and that the biochemical properties and functions of these proteins are diverse. We undertook studies to characterize the biochemical properties of the Drosophila Myb protein (DMyb). We now provide evidence that in addition to having homology with the vertebrate Myb proteins, the Drosophila Myb protein (DMyb) shares its biochemical properties. DMyb binds to a similar consensus sequence and activates transcription from a reporter construct regulated by vertebrate Myb proteins. We also show that DMyb proteins carrying mutations corresponding to previously isolated mutant alleles of Dm myb are less active as transcriptional activators than wild-type DMyb, indicating that a decrease in transcriptional activation ability is likely to cause the mutant phenotypes. PMID- 11500059 TI - Regulation of c-Myb activity by tumor suppressor p53. AB - Heat shock proteins (HSPs) act as chaperones and play important roles during cellular proliferation and apoptosis. Heat shock factors (HSFs) mediate transcriptional induction of HSP genes. Among multiple heat shock transcription factors (HSFs) in vertebrates, HSF3 is specifically activated in unstressed proliferating cells by direct binding to the c-myb proto-oncogene product (c Myb). Since c-Myb has an important role in cellular proliferation, this regulatory pathway suggests a link between the events of cellular proliferation and the stress response. The c-Myb-induced activation of HSF3 is negatively regulated by the p53 tumor suppressor protein. p53 directly binds to HSF3 and blocks the interaction between c-Myb and HSF3. In addition, p53 stimulates the degradation of c-Myb, which is, at least partly, mediated by induction of Siah in certain types of cells. Thus, c-Myb and p53 regulate the expression of HSPs via HSF3 in opposite ways. PMID- 11500060 TI - Telomerase activity in mouse myeloid leukemic cells and in cells from normal hematopoietic systems. AB - The telomeric repeat amplification protocol (TRAP) assay was used to measure telomerase activity in radiation-induced mouse myeloid leukemic (ML) cells and in several populations of normal cells. A detectable level of telomerase activity was found in normal hematopoietic tissues, i.e., bone marrow (BM) cells, day 9 colony-forming unit spleen (CFU-S) colonies, peripheral blood (PB) lymphocytes, and spleen. The level of telomerase activity in normal BM cells was used as a background level. Nine of the 12 cases of ML had higher levels of activity than that of the normal BM cells and therefore they were scored as ML with positive telomerase. The other three cases were considered as ML with negative telomerase because the levels of the enzyme were equivalent to that of normal BM cells. The data indicate that cellular differentiation may suppress telomerase activity in mouse ML cells. In summary, the results suggest that the CBA/Ca mouse model should be a useful animal system for future studies on the assessment of telomerase activity in both malignant and normal hematopoietic cells. PMID- 11500061 TI - Mutations in the HFE gene and their interaction with exogenous risk factors in hepatocellular carcinoma. AB - The possible role of iron in facilitating the development of liver cancer is still debated. The aims of this study were to define the prevalence of the mutations 845G --> A and 187C --> G (C282Y and H63D) in the HFE gene associated with hereditary hemochromatosis in Italian patients with hepatocellular carcinoma occurring in cirrhosis and to analyze the interaction between these mutations and other established risk factors for hepatocellular carcinoma. The HFE gene mutations, performed by polymerase chain reaction, were analyzed in 81 patients (63 males, 18 females) with hepatocellular carcinoma. None of the patients had a phenotype compatible with homozygous hereditary hemochromatosis. Interaction between HFE mutations and exogenous risk factors was analyzed by collecting information on alcohol consumption, hepatitis B and C virus infections, and iron status at the time of diagnosis of chronic liver disease. This analysis was performed only in males to rule out gender influence on patients' iron status by using the case-only approach specifically designed to estimate departure from multiplicative risk ratios under the assumption of independence between genotype and environmental exposure. The prevalence of the C282Y mutation was significantly higher in patients with hepatocellular carcinoma than in normal controls (8.6% vs 1.6%, P < 0.03). At univariate analysis, iron overload was significantly associated with both HFE mutations (P < 0.0001), whereas ongoing hepatitis B virus infection was associated with the C282Y mutation (P < 0.05). By multivariate analysis, a trend for an increased risk of being positive for hepatitis virus markers (OR 2.9, CI 95% 0.9-9.5) and of having been alcohol abusers (OR 3, CI 95% 0.7-14) was observed in patients heterozygous for the HFE mutations. These data indicate that the prevalence of the main mutation associated with hereditary hemochromatosis is significantly higher in cirrhotic Italian patients with hepatocellular carcinoma compared to a normal population and suggest that heterozygotes for HFE mutations exposed to hepatitis virus infections or who had been alcohol abusers could have an increased risk of developing cirrhosis and later liver cancer than people without the mutations exposed to the same risk factors. PMID- 11500062 TI - A new type of inherited catalase deficiencies: its characterization and comparison to the Japanese and Swiss type of acatalasemia. AB - Thirteen Hungarian families that exhibited inherited catalase deficiencies have been detected. Differences between the deficiencies reported from Hungary and the previously reported Swiss acatalasemia were characterized using biochemical analysis of the catalase proteins. Molecular biological methods were used to compare the previously reported types of catalase deficiencies in Japan and the Hungarian deficiencies. Three mutations (a GA insertion in exon 2, a G insertion in exon 2, and a T to G substitution in intron 7) are responsible for decreased catalase activity in 7 of the 13 Hungarian kindreds; the other 6 families have not yet been characterized. These are not the mutations observed in Japan. Changes in lipid and carbohydrate metabolism and the high incidence (12.7%) of diabetes mellitus in the Hungarian kindreds suggest that individuals with inherited catalase deficiency are at risk of atherosclerosis and diabetes mellitus. The Hungarian subjects were detected during screening of a large population for catalase activity; no overt disease state was associated with the deficiencies. We hypothesize that the increased risk of disease may be due to prolonged exposure to elevated levels of blood hydrogen peroxide due to the lack of normal removal of hydrogen peroxide by blood catalase. PMID- 11500063 TI - Low penetrant hemochromatosis phenotype in eight families: no evidence of modifiers in the MHC region. AB - The gene responsible for hemochromatosis (HFE) has been identified on the short arm of chromosome 6, 4.5 Mb telomeric to HLA-A. A major mutation C282Y is closely correlated with the disease, as it accounts for 68 to 100?% of the cases of hemochromatosis. Nevertheless, some C282Y homozygotes subjects have no clinical or biological expression of the disease. Moreover, in Northern European populations a large discrepancy is observed between the number of C282Y homozygotes and the number of diagnosed hemochromatosis patients, suggesting incomplete penetrance of the mutation. To localize and identify the modifying genes, we investigated eight families including C282Y homozygous relatives showing no clinical signs of the disease, in addition to the hemochromatosis patients. Genomic DNA from 20 C282Y homozygotes (10 patients and 10 siblings presenting no or minor biological abnormalities) were studied. Five polymorphisms from the HFE gene were determined by PCR restriction. Extended haplotypes of the 6p21.3 region were constructed with 10 microsatellite markers. All the C282Y homozygotes shared the same HFE polymorphism. The haplotypes presented no significant difference between the probands and their unaffected relatives. These studies suggest that neither HFE polymorphism nor genes surrounding HFE are able to modulate HFE expression. PMID- 11500064 TI - Targeted correction of the point mutations of beta-thalassemia and targeted mutagenesis of the nucleotide associated with HPFH by RNA/DNA oligonucleotides: potential for beta-thalassemia gene therapy. AB - An RNA/DNA chimeric oligonucleotide was found to be effective in the targeted correction of point mutations in Escherichia coli, plant, and mammalian genomes. This strategy, named chimeraplasty, has the potential for gene therapy of many genetic diseases caused by point mutations. beta-Thalassemia is a very common human genetic disease and in most cases it is caused by point mutations. To test whether the chimeraplasty can be used to correct the point mutations responsible for beta-thalassemia, we introduced one mutated beta-globin gene, betaE, into MEL cells and successfully corrected the point mutation of the betaE gene with the highest correction efficiency of 1.9%. Furthermore, a targeted -202 C-->G mutation of the Ggamma-globin gene, which is associated with the elevated Ggamma globin gene expression in the adult stage, was introduced into HeLa and CMK cells by an RNA/DNA oligonucleotide. These results indicated that the chimeraplasty has potential for human beta-thalassemia gene therapy. PMID- 11500065 TI - Polymorphisms in the transferrin 5' flanking region associated with differences in total iron binding capacity: possible implications in iron homeostasis. AB - We have identified five single nucleotide polymorphisms (SNPs) upstream (5') of the transferrin coding region. One polymorphism is in the 5' UTR at nt +49, and four are in the promoter region at nt -34, -551, -617, and -739, numbering from the start of transcription. The -34 and -617 SNPs are tightly but not completely linked. The -34 polymorphism lies between a conserved Sp1 site and the TATA box. The -617 polymorphism is within the DRII enhancer region. Five haplotypes have been defined from these SNPs by the identification of at least one homozygous individual, and two other haplotypes were deduced from heterozygous individuals. The total iron-binding capacity associated with each transferrin haplotype was haplotype 2 > 1 > 4 > 3. Transferrin promoter haplotype 2 had a significantly higher mean TIBC and haplotype 3 had a significantly lower mean TIBC than the more common haplotype 1. Persons with haplotype 4, which includes the -34T and 617A minor alleles, have a lower mean TIBC but the difference was not statistically significant. In normal individuals, the differences in the haplotypes were not found to be associated with differences in transferrin saturation and ferritin levels. There was no difference in the extent of increase in the mean TIBC levels in individuals with iron deficiency anemia in regard to their haplotype. Furthermore, there was no difference in the relative frequencies of the transferrin haplotypes in the iron-deficient population. In hemochromatosis patients who were homozygous for the C282Y HFE mutation, no particular haplotype was associated with a significant difference in transferrin saturation or ferritin levels. In White patients with Parkinson's disease, a disorder in which there is abnormal iron deposition in the brain, the presence of transferrin haplotype 3 was in slight excess over the normal White population. PMID- 11500066 TI - Frequency analysis and allele map in favor of the celtic origin of the C282Y mutation of hemochromatosis. AB - After the main hereditary hemochromatosis mutation C282Y in the HFE gene was described, we report here the C282Y frequencies for various European populations. The aim of this meta-analysis is to compile the Y allele frequencies of the C282Y mutation for 53 European populations, representing a total of 9265 unrelated people representing control samples. The most elevated values are observed in residual Celtic populations in Ireland, in the United Kingdom, and in France, in accordance with the initial hypothesis of Simon et al. (Prog. Med. Genet. 4, 135 168, 1980) concerning a Celtic origin of the hereditary hemochromatosis mutation. PMID- 11500067 TI - Walking or talking? Behavioral and neurophysiological correlates of action verb processing. AB - Brain activity elicited by visually presented words was investigated using behavioral measures and current source densities calculated from high-resolution EEG recordings. Verbs referring to actions usually performed with different body parts were compared. Behavioral data indicated faster processing of verbs referring to actions performed with the face muscles and articulators (face related words) compared to verbs referring to movements involving the lower half of the body (leg-related words). Significant topographical differences in brain activity elicited by verb types were found starting approximately 250 ms after word onset. Differences were seen at recording sites located over the motor strip and adjacent frontal cortex. At the vertex, close to the cortical representation of the leg, leg-related verbs (for example, to walk) produced strongest in-going currents, whereas for face-related verbs (for example, to talk) the most in-going activity was seen at more lateral electrodes placed over the left Sylvian fissure, close to the representation of the articulators. Thus, action words caused differential activation along the motor strip, with strongest in-going activity occurring close to the cortical representation of the body parts primarily used for carrying out the actions the verbs refer to. Topographically specific physiological signs of word processing started earlier for face-related words and lasted longer for verbs referring to leg movements. We conclude that verb types can differ in their processing speed and can elicit neurophysiological activity with different cortical topographies. These behavioral and physiological differences can be related to cognitive processes, in particular to lexical semantic access. Our results are consistent with associative theories postulating that words are organized in the brain as distributed cell assemblies whose cortical distributions reflect the words' meanings. PMID- 11500068 TI - Prosodic facilitation in the resolution of syntactic ambiguities in subjects with left and right hemisphere damage. AB - The purpose of this study was to determine if prosody facilitates the comprehension of sentences containing temporary syntactic ambiguities in control, and left (LHD) and right hemisphere damaged (RHD) subjects. To test for effects of prosodic facilitation, sentences were created where prosodic boundaries coincided with (cooperating), were absent (baseline), or conflicted (conflicting) with syntactic boundaries in three response times (RTs) experiments. Despite differences in overall RTs and response accuracy for each group, all three groups responded faster and more accurately to sentences in the cooperating than in the baseline and conflicting conditions across experiments, indicating that prosody facilitates syntactic parsing in brain-damaged subjects just as it does with normal control subjects. Results are discussed in relation to psycholinguistic theories of syntactic parsing and neurolinguistic theories of hemispheric specialization in processing the acoustic properties of prosodic structures. PMID- 11500069 TI - An investigation of the interaction between thematic and phrasal structure in nonfluent agrammatic subjects. AB - Garrett (1982) developed a model of normal sentence production which has been used in the description of aphasic language (Schwartz, 1987). This study investigated the effects of the thematic representation specified at the functional level on the complexity of the phrases produced at the positional level. A group of 14 nonfluent, agrammatic subjects were compared to 20 normal controls in their production of the story of Cinderella. The agrammatic subjects produced fewer argument structures than the normal control subjects. Their phrasal realization of the arguments, however, was not qualitatively different from that of the normal subjects. In both cases, with an increase in the number of arguments, there was a concurrent increase in the mean complexity of the phrases used to realize those arguments and in the total phrasal complexity of the utterances. The complexity of noun phrases differed according to the thematic roles expressed; this seemed to be a consequence of their different locations in the sentence. Preverbal noun phrases were much less complex than postverbal noun phrases. There was no evidence to suggest that there was a trade-off between the production of thematic structure and subsequent phrasal production. Neither was there evidence to suggest that production differed according to whether the phrase was an argument of the verb or a nonargument. The complexity of a phrase was determined by the type of information it conveyed. PMID- 11500070 TI - From "logographic" to normal reading: the case of a deaf beginning reader. AB - Visual word recognition of a profoundly deaf girl (AH) with developmental reading disorders was explored using an experimental technique that measures performance as a function of eye fixation within a word. AH's fixation-dependent word recognition profile revealed that she was inferring the identity of words using a "logographic" reading strategy (i. e., using salient visual features). Following this observation a special training program that enhances the understanding of grapheme-phoneme relations was applied. After few months of training, AH's reading skills improved, while her fixation-dependent performance changed to become like that of normal readers. We discuss the impact of our technique for the early diagnosis of reading impairments. PMID- 11500071 TI - Sources of performance on theory of mind tasks in right hemisphere-damaged patients. AB - Patients with right hemisphere (RHD) or left hemisphere brain damage (LHD) were tested on Theory of Mind (ToM) tasks presented with visual aids that illustrated the relevant premises. As a measure of pragmatic ability, patients were also asked to judge replies in conversation that violated Gricean maxims. Both RHD and LHD patients performed well on the ToM tasks presented with visual aids, but RHD patients displayed difficulty when the same tasks were presented only verbally. In addition, RHD patients showed reduced sensitivity to pragmatic violations. These findings point to the role of right hemisphere structures in processing information relevant to conversations. They indicate that a crucial source of RHD patients' errors in ToM tasks may involve difficulties in utterance interpretation owing to impairments of visuospatial processing required for the representation of textual information. PMID- 11500072 TI - Effect of bilateral stimulation of the subthalamic nucleus on parkinsonian voice. AB - The purpose of this study was to assess several acoustic features of the voices of 26 parkinsonian patients under two conditions, with and without bilateral chronic stimulation of the subthalamic nucleus (STN) to estimate the effectiveness of this procedure on parkinsonian speech. When compared to unstimulated patients, stimulated patients showed longer duration of sustained vowels, shorter duration of sentences, nonsense words, and pauses, more variable fundamental frequency (f0) in sentences, and more stable f0 during sustained vowels. Relative intensity was unchanged in both conditions. Further acoustic analyses are warranted to clarify the role of STN stimulation on parkinsonian speech. PMID- 11500073 TI - Neurological evidence in support of a specialized phonetic processing module. AB - Event-related potentials (ERPs) were utilized to study brain activity while subjects listened to speech and nonspeech stimuli. The effect of duplex perception was exploited, in which listeners perceive formant transitions that are isolated as nonspeech "chirps," but perceive formant transitions that are embedded in synthetic syllables as unique linguistic events with no chirp-like sounds heard at all (Mattingly et al., 1971). Brain ERPs were recorded while subjects listened to and silently identified plain speech-only tokens, duplex tokens, and tone glides (perceived as "chirps" by listeners). A highly controlled set of stimuli was developed that represented equivalent speech and nonspeech stimulus tokens such that the differences were limited to a single acoustic parameter: amplitude. The acoustic elements were matched in terms of number and frequency of components. Results indicated that the neural activity in response to the stimuli was different for different stimulus types. Duplex tokens had significantly longer latencies than the pure speech tokens. The data are consistent with the contention of separate modules for phonetic and auditory stimuli. PMID- 11500074 TI - Effect of socioeconomic status on aphasia severity and recovery. AB - Low levels of educational attainment and low socioeconomic status have been significantly linked to poor health and increased incidence of disease, including Alzheimer's disease and diseases of the cardiovascular, pulmonary, renal, and gastrointestinal systems. Our goal in the present study was to determine the degree to which educational level and socioeconomic status influence initial severity of aphasia and subsequent recovery. We evaluated the records of 39 persons with aphasia twice: at about 4 months and 103 months postonset. We found early severity of aphasia to be significantly greater for subjects in the lower educational and occupational groups. However, rate of recovery (the slope of the recovery curve) was the same regardless of educational or occupational status. PMID- 11500075 TI - The frequency paradox in disguise: a response to Breedin, Saffran, and Schwartz (1998). PMID- 11500077 TI - A tale of two paradigms or metatheoretical approaches to cognitive neuropsychology: did Schmolck, Stefanacci, and Squire (2000) show that hippocampal lesions only impair memory, whereas adjacent (extrahippocampal) lesions impair detection and explanation of sentence ambiguity? AB - This note discusses two fundamentally different paradigms or metatheoretical approaches that currently guide cognitive neuropsychology: the Theoretical- vs. Anatomical-paradigms. To illustrate these paradigms, we compare a Theoretical paradigm paper (MacKay & James, 2001) with an Anatomical-paradigm paper (Schmolck, Stefanacci, & Squire, 2000): These papers report virtually identical experiments on relations between language, memory, and hippocampal systems, using the same task (the detection and explanation of ambiguities in sentences that participants know are ambiguous), virtually identical ambiguous sentences, and at least one identical participant (the amnesic HM). However, MacKay and James made strikingly different claims from Schmolck et al., and we show that the Schmolck et al. claims comport not with their data but with an unstated theory to which they are implicitly committed within the Anatomical-paradigm. PMID- 11500080 TI - Detection, characterisation and purification of a murine liver factor capable of desensitising towards the lethal activity of tumour necrosis factor. AB - Tumour necrosis factor (TNF) is a major mediator in septic shock and several inflammatory diseases such as hepatitis. Galactosamine (GalN) sensitises experimental animals for TNF and the combination TNF/GalN leads to a lethal inflammatory hepatitis. We describe that a single injection of lipopolysaccharide (LPS), interleukin-1 (IL-1) or TNF can desensitise against the lethality induced by TNF/GalN, but also against changes in metabolic parameters such as hypothermia and transaminase release, in a dose responsive way. We also describe the desensitising capacity of a component present in Mouse Liver Extract (MLE). The MLE desensitises mice against the effects of TNF/GalN in a dose responsive way. The activity of the MLE is heat labile and does not involve LPS, TNF, IL-1 or TNF soluble receptors. We describe partial and complete purification of the factor. Partially pure material protects mice against all changes induced by TNF/GalN. The protection is dose dependent and heat labile and also possible in endotoxin hyporesponsive C3H/HeJ mice. The pure material protects against lethality, hypothermia and AST release and it appears as a heat labile protein of relative molecular weight of 70 kDa probably with a break down product of 35 kDa. PMID- 11500081 TI - Reduced expression of flice-inhibitory protein (FLIP) and NFkappaB is associated with death receptor-induced cell death in human aortic endothelial cells (HAECs). AB - We investigated the effects of TNF receptor 1 (TNFR1) and 2 (TNFR2) modulation on the death of human aortic endothelial cells (HAECs) resistant to TNF-alpha induced cell death. Alteration of the transcription of anti-apoptotic proteins, including inhibitor of apoptosis protein 1, 2 (cIAP1, 2), TNF receptor-associated factor 1 (TRAF1), nuclear factor kappa B1 protein (NFkappaB1), and FLICE inhibitory protein (FLIP) was assessed by real-time reverse transcriptase polymerase chain reaction (RT-PCR). TNF-alpha (200 ng/ml) or actinomycin D (ActD) (5 ng/ml) did not kill cells, while 5 ng/ml of TNF-alpha and 5 ng/ml of ActD increased expression of Fas (CD95) and FasL (CD95L), and 45% of cells died. TNFR2 blockade suppressed NFkappaB1 and FLIP expression and increased cell death. TNFR1 blockade enhanced FLIP expression and decreased cell death. Cells insensitive to TNF-alpha may respond to TNF-alpha through TNFR that induces transcription of NFkappaB1 and FLIP. Down-regulation of these proteins may facilitate death of cells insensitive to TNF-alpha-induced cell death. PMID- 11500082 TI - Evidence for a novel thrombopoietin signalling event: activation of protein kinase A in human megakaryoblastic CMK cells. AB - Thrombopoietin (TPO) plays a crucial role in megakaryocyte development. TPO signalling, which is mediated by its receptor Mpl, includes Janus kinase, (JAK) signal transducer and activator of transcription (STAT) and Shc/Ras/mitogen activated protein kinase (MAPK) pathways. The precise nature of these signalling routes has not been clarified in detail up until now. We investigated the effect of TPO on activation of cAMP-dependent protein kinase (PKA) and its involvement in MAPK signalling in human megakaryoblastic leukaemia CMK cells. For estimation of PKA activity, phosphorylation of a PKA-specific peptide substrate was assayed in CMK cell lysates. Since activation of PKA is associated with translocation of its catalytic subunit alpha (C-PKA) into the cell nucleus, Western blot analysis of nuclear fractions with an anti-C-PKA antibody was additionally performed. The activation of TPO-induced MAPK activation and the effect of the PKA inhibitor H 89 was measured using immunoblotting with a monoclonal anti-pERK antibody. TPO enhanced cAMP and induced activation of PKA in CMK cells. In addition, H-89 partly blocked TPO-induced MAPK activation in CMK cells. Our results indicate a novel TPO-triggered signalling event, activation of the cAMP/PKA pathway in human megakaryoblastic CMK cells. This signal transduction route seems to be involved in TPO-induced MAPK signaling. PMID- 11500083 TI - High tumour necrosis factor-alpha levels are associated with exercise intolerance and neurohormonal activation in chronic heart failure patients. AB - Immune activation plays an important role in the progression of chronic heart failure (CHF). We sought to investigate whether different degrees of tumor necrosis factor-alpha (TNF-alpha) activation are associated with exercise intolerance, neurohormonal activation and alterations in muscle mass and function in patients with CHF without cardiac cachexia. Patients were divided into quartiles according to their TNF levels (first quartile: 0.98-4.90 pg/ml, second quartile: 5.00-6.60 pg/ml; third quartile 6.80-9.00 pg/ml; fourth quartile 9.80 32.00 pg/ml). Patients underwent cardiopulmonary exercise testing, quadriceps muscle strength test, quadriceps fatigue test, and assessment of thigh muscle and fat cross-sectional area (CSA) by computerized tomography scanning. Patients in the highest TNF quartile had the lowest peak oxygen consumption [13.1 (+/-4.1) ml/kg/min vs 18.1 (+/-5.3), 18.8 (+/-4.8) and 18.7 (+/-5.6) ml/kg/min, P<0.01] the greatest relation of ventilation and dioxide production (VE/VCO(2)) slope (P<0.05) and the most elevated catecholamine levels (P<0.05) compared to patients in the first three quartiles. Patients with the lowest TNF levels had preserved thigh muscle size and quadriceps strength. Strength/muscle CSA was similar in the four groups. Muscle strength during fatigue testing was significantly lower in the fourth quartile (P=0.01) compared with the other three groups. In CHF patients only the highest levels of TNF are associated with poor functional status and neurohormonal activation. This group of patients may represent the appropriate target population for TNF antagonism. PMID- 11500084 TI - Recombinant human macrophage colony-stimulating factor augments pulmonary host defences against Aspergillus fumigatus. AB - The in vivo and ex vivo effects of macrophage colony-stimulating factor (M-CSF) were studied in a profoundly neutropenic rabbit model in order to determine its potential to augment pulmonary host defence against Aspergillus. M-CSF (100-600 microg/kg/d) was administered prophylactically to neutropenic rabbits with pulmonary aspergillosis starting three days pre-inoculation and then throughout neutropenia. Rabbits receiving M-CSF had significantly increased survival (P=0.01) and decreased pulmonary injury, as measured by decreased pulmonary infarction (P=0.004), when compared with untreated controls. Microscopic studies demonstrated greater numbers of activated pulmonary alveolar macrophages (PAMs) in lung tissue of rabbits receiving M-CSF, in comparison to controls (P<0.001). PAMs harvested from rabbits treated with M-CSF had a significantly greater percent phagocytosis of Aspergillus fumigatus conidia than did PAMs from controls (P=0.04). These data indicate that prophylactic administration of M-CSF augments pulmonary host defence against A. fumigatus and suggest a potential role for this cytokine as adjunctive therapy in the treatment of pulmonary aspergillosis in the setting of profound neutropenia. PMID- 11500085 TI - Role of CD23 in astrocytes inflammatory reaction during HIV-1 related encephalitis. AB - Soluble factors released by intra-cerebral activated cells are implicated in neuronal alterations during central nervous system inflammatory diseases. In this study, the role of the CD23 pathway in astrocyte activation and its participation in human immunodeficiency virus-1 (HIV-1)-induced neuropathology were evaluated. In human primary astrocytes, CD23 protein membrane expression was dose dependently upregulated by gp120. It was also upregulated by gamma-interferon (gamma-IFN) and modulated by interleukin-1-beta (IL-1beta) whereas microglial cells in these stimulation conditions did not express CD23. Cell surface stimulation of CD23 expressed by astrocytes induced production of nitric oxide (NO) and IL-1beta which was inhibited by a specific inducible NO-synthase (iNOS) inhibitor (aminoguanidine), indicating the implication of this receptor in the astrocyte inflammatory reaction. On brain tissues from five out of five patients with HIV-1-related encephalitis, CD23 was expressed by astrocytes and by some microglial cells, whereas it was not detectable on brain tissue from five of five HIV-1-infected patients without central nervous system (CNS) disease or from two of two control subjects. In addition, CD23 antigen was co-localized with iNOS and nitrotyrosine on brain tissue from patients with HIV1-related encephalitis, suggesting that CD23 participates in iNOS activation of astrocytes in vivo. In conclusion, CD23 ligation is an alternative pathway in the induction of inflammatory product synthesis by astrocytes and participates in CNS inflammation. PMID- 11500086 TI - TNF and CD95 promote IL-8 gene transactivation via independent elements in colon carcinoma cells. AB - The pro-inflammatory cytokine interleukin-8 (IL-8) is produced by HT29 colon epithelial cells following engagement of either CD95 or tumour necrosis factor (TNF) receptors. While the IL-8 promotor elements activated by TNF are well characterised, those responsible for induction of IL-8 by CD95 are unknown. We examined the pathway for CD95 induced IL-8 secretion using two luciferase reporter constructs; the first comprising approximately 500 bp of the IL-8 promotor that includes the nuclear factor kappa B (NFkappaB), C/EBP and AP-1 sites known to be involved in TNF mediated IL-8 induction; the second that encompasses these elements but extends approximately 1.1 kb further upstream. Although IL-8 mRNA and protein were produced in response to either TNF or CD95 ligation, only TNF induced an increase in the reporter activity of the promoter constructs. Nevertheless, IL-8 induction by CD95 resulted primarily from increased transcription and not from an increase in IL-8 mRNA stability. These results suggest that promoter elements/enhancers involved in CD95 mediated IL-8 induction are distinct from those used by TNF and not contained within the 1.6 kb region immediately upstream of the initiation codon. PMID- 11500087 TI - Adult survival, maturation, and reproduction of the desert locust Schistocerca gregaria infected with the fungus Metarhizium anisopliae var acridum. AB - Studies were conducted with two different doses of Metarhizium anisopliae var acridum to examine the effects on survival and reproductive potential of adult Schistocerca gregaria under conditions that either limited thermoregulation or enabled optimal thermoregulation. Adult S. gregaria infected with the fungal pathogen showed either a rapid and high mortality at relatively constant temperatures or a much reduced mortality and lengthened survival time when allowed to thermoregulate. Mortality rate varied from >90% after 10 days under constant temperature conditions to 66% after 70 days under optimal thermoregulatory conditions. Effects of infection on maturation and reproduction depended on the age of the adults at the time of inoculation, the nighttime temperature regime, the fungal dose, and the length of time of the monitoring period. No difference in reproductive behaviors in treated and control insects were found in one experiment that utilized older adults and was conducted over 25 days. In a second experiment with newly fledged locusts, differences in maturation rates and total reproductive output were observed due to infection. The results from these experiments are discussed in terms of the potential of M. anisopliae var acridum to alter the balance of insect endocrine systems and the importance of the assessment of behavioral changes and their impact on microbial control agents in the long term. PMID- 11500088 TI - Epidemiological parameters of White Spot Syndrome Virus infections in Litopenaeus vannamei and L. setiferus. AB - An experimental protocol based on a mathematical epidemiology model was developed to study the transmission, virulence, and recovery rates of White Spot Syndrome Virus (WSSV). Two modes of transmission were compared for WSSV in Litopenaeus vannamei. We compared transmission by ingestion of infected cadavers to transmission by cohabitation with infected animals. In addition, we compared the ingestion transmission of WSSV in L. vannamei and in L. setiferus. Finally, we compared the virulence and recovery rates of WSSV in L. vannamei and L. setiferus. The transmission rate of WSSV to L. vannamei by cohabitation was 0.01. The transmission rate by ingestion of infected cadavers was over an order of magnitude larger at 0.46, suggesting that cohabitation is a much less important mode of transmission for WSSV. A statistically significant difference was detected between the estimates of ingestion transmission of L. vannamei (0.46) and those of L. setiferus (0.84), yet no differences in the virulence or recovery rates were detected between hosts. The overall estimated virulence rate was 0.34, and the overall estimated recovery rate from a WSSV infection was 0.007 for both species. According to epidemiological theory the threshold density of hosts necessary for an epidemic to occur is directly related to the virulence and recovery rates and inversely related to the transmission rate. Therefore, the epidemic threshold density may be lower for ingestion transmission than cohabitation transmission and lower for L. setiferus than for L. vannamei. PMID- 11500089 TI - Formulation of entomopathogenic nematode-infected cadavers. AB - Entomopathogenic nematodes are commercially applied in aqueous suspension. These biocontrol agents may also be applied in nematode-infected insect cadavers, but this approach may entail problems in storage and ease of handling. We determined the feasibility of formulating nematode-infected insect cadavers to overcome these hindrances. All experiments were conducted with Heterorhabditis bacteriophora Poinar and Galleria mellonella (L.). Nonformulated cadavers were used as controls. Of 19 formulations tested (including combinations of starches, flours, clays, etc.) 1 (starch-clay combination) was found to adhere to the cadaver and to have no significant deleterious effects on nematode reproduction and infectivity; other formulations exhibited poor adhesion or reduced nematode reproduction. Two formulations enabled cadavers to be partially desiccated without affecting reproduction; other formulations and nonformulated cadavers exhibited reduced reproduction upon desiccation. Four-day-old cadavers were more amenable to desiccation than 8-day-old cadavers. Formulated cadavers were more resistant to rupturing and sticking together during agitation than nonformulated cadavers. PMID- 11500090 TI - A bacterially produced virus enhancing factor from an entomopoxvirus enhances nucleopolyhedrovirus infection in armyworm larvae. AB - Using an Escherichia coli expression system, pGEX-2T, that expresses foreign sequences as fusion proteins with a glutathione S-transferase (GST) carrier, we have expressed a virus enhancing factor (EF) from Pseudaletia separata entomopoxvirus, which enhances P. unipuncta multi nucleopolyhedrovirus (PsunMNPV) infection in larvae of the armyworm, P. separata. The lysates of transformed E. coli cells, which were not active in enhancing PsunMNPV infection, became active when treated with either trypsin or thrombin. The GST-EF fusion protein in a lysate was purified with a bulk GST purification module and cleaved into the EF and GST moieties with thrombin. Removal of the GST moiety with glutathione Sepharose 4B resulted in a highly purified EF preparation, which enhanced PsunMNPV infection in armyworm larvae and PsunMNPV fusion with an armyworm cell line, SIE-MSH-805-F. PMID- 11500091 TI - Factors influencing the infectivity of isolates of Paecilomyces fumosoroseus against diamondback moth, Plutella xylostella. AB - Paecilomyces fumosoroseus isolate 1576 was isolated from an insect, but is avirulent against larvae of diamondback moth, Plutella xylostella, and several other species. Isolate 1576 grew faster and produced more conidia than isolate 4461 on potato dextrose agar. Pregermination of conidia failed to increase the infectivity of isolate 1576, but the procedure did increase the infectivity of isolates 3682, 4461, and 4482. Isolates 1576 and 4461 were both more infective when moisture was high during incubation of inoculated larvae. Starved Pl. xylostella larvae were more susceptible than fed larvae to isolate 1576 (40 and 10% mortality, respectively), but starved and fed larvae were similar in susceptibility to isolate 4461. These results show that isolate 1576 grows vigorously in aerial culture and is capable of infecting stressed Pl. xylostella larvae. Further tests are needed to characterize its pathogenicity toward its original host or closely related species. PMID- 11500092 TI - Construction and characterization of a recombinant Bacillus thuringiensis subsp. israelensis strain that produces Cry11B. AB - The mosquitocidal bacterium Bacillus thuringiensis subsp. israelensis (Bti) produces four major endotoxin proteins, Cry4A, Cry4B, Cry11A, and Cyt1A, and has toxicity in the range of many synthetic chemical insecticides. Cry11B, which occurs naturally in B. thuringiensis subsp. jegathesan, is a close relative of Cry11A, but is approximately 10-fold as toxic to Culex quinquefasciatus. To determine whether the addition of Cry11B to Bti would improve its toxicity, we produced this protein in Bti. High levels of Cry11B synthesis were obtained by expression of the cry11B gene under the control of cyt1A promoters and the STAB SD sequence. This construct was cloned into the shuttle vector pHT3101, yielding the derivative plasmid pPFT11Bs, which was then transformed by electroporation into acrystalliferous (4Q7) and crystalliferous (IPS-82) strains of Bti. Synthesis of Cry11B in Bti 4Q7 produced crystals approximately 50% larger than those produced with its natural promoters without STAB-SD. However, less Cry11B was produced per unit culture medium with this construct than with the wild-type construct, apparently because the latter construct produced more cells per unit medium. Nevertheless, the Bti IPS-82 strain that produced Cry11B with pPFT11Bs was twice as toxic as the parental IPS-82 strain (LC(50) = 1.4 ng/ml versus 3.3 ng/ml, respectively) to fourth instars of C. quinquefasciatus. Against fourth instars of Aedes aegypti, no statistically significant difference between parental Bti IPS-82 (LC(50) = 4.7 ng/ml) and the Bti IPS-82 recombinant producing Cry11B (LC(50) = 3.5 ng/ml) was found in toxicity. PMID- 11500093 TI - Gene knockout demonstrates that vip3A contributes to the pathogenesis of Bacillus thuringiensis toward Agrotis ipsilon and Spodoptera exigua. AB - Vip3A is an 89-kDa protein secreted by Bacillus thuringiensis during vegetative growth. To determine the importance of Vip3A for the insect pathogenicity of B. thuringiensis the vip3A gene was deleted from strain HD1, yielding strain HD1Deltavip3A. Compared with HD1, strain HD1Deltavip3A was one-fourth as toxic to Agrotis ipsilon larvae and less than one-tenth as toxic to Spodoptera exigua larvae. When streptomycin was included in the S. exigua diet the toxicity of HD1Deltavip3A was approximately half that of HD1. Addition of HD1 spores increased the toxicity of purified Cry1 protein more than 600-fold against S. exigua, whereas addition of HD1Deltavip3A spores increased toxicity of Cry1 protein approximately 10-fold. These results demonstrate that an important component of B. thuringiensis insecticidal activity against S. exigua is the synthesis of Vip3A protein by B. thuringiensis cells after ingestion of spores and crystal proteins by insect larvae. PMID- 11500094 TI - Gender differences in phenoloxidase activity of Panorpa vulgaris hemocytes. PMID- 11500095 TI - The Cry toxins and the putative hemolysins of Clostridium bifermentans ser. malaysia are not involved in mosquitocidal activity. PMID- 11500097 TI - Characterization of the CsfC and CsfD proteins involved in the biogenesis of CS5 pili from enterotoxigenic Escherichia coli. AB - The region required for biosynthesis of CS5 pili consists of six csf genes, with csfA encoding the major subunit. In this study, we describe the characterization of two of the genes constituting the region, csfC and csfD, but also identify the true morphology of the CS5 pilus by high resolution electron microscopy. CsfD was shown to be essential in the initiation of CS5 pilus biogenesis, did not possess any chaperone-like activity for the major subunit, and was an integral minor component of the pilus structure. Studies on CsfD translocation across the outer membrane in Escherichia coli K-12 using a csfA mutant also showed that CsfD is likely to be the first pilin subunit assembled. A specific in-frame deletion in the csfC gene resulted in the complete absence of cell surface CS5 pili and prevented the translocation of CsfA and CsfD pilins across the outer membrane. Specific cell localization studies showed an accumulation of CsfC in the outer membranes of E. coli K-12, while complementation experiments with homologous outer membrane assembly genes from CS1 and CFA/I pili systems were unable to restore assembly of CS5 pili. The CS5 pilus was shown to be a 2 nm flexible fibrillar structure, which adopted a predominantly open helical conformation under the electron microscope. PMID- 11500096 TI - Complementation of a speA negative Streptococcus pyogenes with speA: effects on virulence and production of streptococcal pyrogenic exotoxin A. AB - We have shown previously that an isogenic SPEA-negative Streptococcus pyogenes strain did not attenuate virulence in a murine model of necrotizing fasciitis. The aim of this study was to confirm that streptococcal pyrogenic exotoxin A (SPEA) is not crucial for streptococcal invasiveness in murine invasive infection. The SPEA-negative S. pyogenes (H326) was complemented with speA extra chromosomally to create strain H361 which produced 2.2-fold more SPEA compared with the parental speA(+)wild-type (H305). The growth phase-regulated expression of SPEA in vitro was unaffected in this strain. Complementation with speA resulted in reduced virulence and bacterial counts in invasive murine infection. SPEA production was quantitated from muscle tissue of infected mice. However, H361 did not produce more SPEA than H305 in vivo. We conclude that SPEA does not play a key role in invasive murine streptococcal infection. PMID- 11500099 TI - Orientia tsutsugamushi suppresses the production of inflammatory cytokines induced by its own heat-stable component in murine macrophages. AB - Orientia tsutsugamushi is a Gram-negative obligate intracellular bacterium, which causes scrub typhus. To understand the pathogenesis of scrub typhus, we have investigated the induction of tumor necrosis alpha (TNF-alpha) and interleukin-6 (IL-6) by O. tsutsugamushi in two murine macrophage cell lines. Both live and heat-killed orientia stimulated the production of cytokines in J774A.1 cells. Polymyxin B does not affect the secretion of cytokines. These together with the fact that the immature macrophage cell line, P388D1, did not produce TNF-alpha when induced by either live or heat-killed O. tsutsugamushi strongly argue against any roles of lipopolysaccharide (LPS) in cytokine production. Furthermore, the result that the cytokine responses were more brisk when macrophage cell lines had been induced by heat-killed O. tsutsugamushi than by live organisms strongly suggest that a heat-stable molecule might be responsible for the induction of cytokine production and O. tsutsugamushi might have mechanisms suppressing the production of inflammatory cytokines induced by its own heat-stable molecule. PMID- 11500098 TI - Binding of fibronectin by Trichomonas vaginalis is influenced by iron and calcium. AB - We have reported that Trichomonas vaginalis, the causative agent of the most common, non-viral sexually transmitted disease, bound to cover slips coated with fibronectin (FN) (Crouch & Alderete, Microbiol 1999 145: 2835-43). In this study, we extend that observation by showing that FN binding is specific, and we present data on the requirements of FN binding by T. vaginalis. Immunofluorescence and immuno-gold labelling readily detected FN throughout the trichomonal surface. Parasites bound to(125)I-labelled FN in a time- and concentration-dependent fashion. In the absence of protease inhibitor, iodinated FN was released from the trichomonad surface. Unlabelled FN specifically competed for binding in a concentration-dependent fashion with the(125)I-labelled FN. Interestingly, the amount of FN bound by T. vaginalis organisms was dependent on iron. High-iron grown trichomonads acquired lower numbers of molecules but with 10-fold higher affinity than low-iron-grown organisms. Further, we show that for iron-replete organisms, calcium (Ca(2+)) at physiological levels increased amounts of bound FN. The increase in binding was rapid, occurring within 5 min of Ca(2+)addition, and required de novo protein synthesis. Co-incubation of live parasites with Ca(2+)in the presence of FN was necessary to increase the amount of FN bound. Treatment of trichomonads with okadaic acid, but not other phosphatase inhibitors, resulted in a 50% decrease in binding of FN, regardless of the presence of Ca(2+), suggesting a role for phosphatase in FN association. These results indicate that depending on the iron status of T. vaginalis organisms in vivo, Ca(2+)may influence trichomonad recognition and binding to FN during host parasitism. PMID- 11500103 TI - Topological indices based on vertex, distance, and ring: on the boiling points of paraffins and cycloalkanes. AB - Vertex, distance, and ring (in cyclic compounds) are three essential structure elements of a molecular graph, based on which three new topological indices VDI, OEI, and RDI are proposed. Multiple regression analysis was carried out against the boiling points of 343 hydrocarbons(160 paraffins and 183 cycloalkanes) with VDI, OEI, RDI, and N(2/3)(N is the number of carbon atoms) together, and a good QSPR model was obtained: ln(577 - Bp) = 6.729609 - 0.154107N(2/3) + 2.285632 x 10(-2)VDI - 7.921410 x 10(-3)OEI - 1.821962 x 10(-2)RDI (F = 6455.09, r = 0.9935, rms = 6.44 degrees C, n = 343). PMID- 11500100 TI - Protein D expression promotes the adherence and internalization of non-typeable Haemophilus influenzae into human monocytic cells. AB - Protein D, having a glycerol-3-phosphodiester phosphodiesterase activity, is found at the surface of all Haemophilus influenzae strains and is a possible virulence factor. In the present study, the involvement of protein D in the entry of NTHi into human monocytic cells is reported. Primary monocytes and the monocytic cell lines U-937 and THP-1 were infected with NTHi strain 772 and the mutant 772 Delta hpd 1 (lacking the gene for protein D). NTHi 772 adhered to and entered monocytic cells up to four-fold more efficiently compared to 772 Delta hpd 1. When an Escherichia coli transformant expressing protein D was incubated with monocytic cells, the number of intracellular bacteria increased 1.6-fold compared to protein D-deficient controls. Any correlation between internalization and phosphorylcholine expression was not detected. In conclusion, our data suggest that surface-expressed protein D promotes the adherence of NTHi to human monocytes leading to a higher number of internalized bacteria. PMID- 11500104 TI - A bibliometric study on chemical information and computer sciences focusing on literature of JCICS. AB - A bibliometric approach was used to survey the state-of-the-art of research in the field of chemical information and computer sciences (CICS). By examining the CA database for the articles abstracted under the subsection "Chemical information, documentation, and data processing", Journal of Chemical Information and Computer Sciences (JCICS) was identified to have been the top journal in this subsection for the last 30 years. Based on this result, CA subsections and controlled index terms given to JCICS articles were analyzed to see trends in subjects and topics in the CICS field during the last two decades. These analyses revealed that the subjects of research in CICS have diversified from traditional information science and computer applications to chemistry to "molecular information sciences". The SCISEARCH database was used to grasp interdependency between JCICS and other key journals and also the international nature of JCICS in its publications and citedness. PMID- 11500105 TI - A novel formalism to characterize the degree of unsaturation of organic molecules. AB - The existing formalism to calculate the degree of unsaturation from the molecular formula of organic molecules cannot be applied to charged and/or disconnected species. Moreover, the calculated value depends on the assumed formal valence of each of the elements. In this work, we introduce a new formalism that eliminates these problems. The suggested property, degree of unsaturation, can be calculated from the molecular formula as well as from any structural representation of a molecule corresponding to that molecular formula. PMID- 11500106 TI - Stochastic generator of chemical structure. 3. Reaction network generation. AB - A new method to generate chemical reaction network is proposed. The particularity of the method is that network generation and mechanism reduction are performed simultaneously using sampling techniques. Our method is tested for hydrocarbon thermal cracking. Results and theoretical arguments demonstrate that our method scales in polynomial time while other deterministic network generators scale in exponential time. This finding offers the possibility of investigating complex reacting systems such as those studied in petroleum refining and combustion. PMID- 11500107 TI - A dissipative exponentially-fitted method for the numerical solution of the Schrodinger equation. AB - A dissipative exponentially fitted method is constructed in this paper for the numerical integration of the Schrodinger equation. We note that the present method is a nonsymmetric multistep method (dissipative method) An application to the bound-states problem and the resonance problem of the radial Schrodinger equation indicates that the new method is more efficient (i.e. more accurate and more rapid) than the classical dissipative method and other well-known methods. Based on the new method and the method of Raptis and Allison(19) a new variable step method is obtained. The application of the new variable-step method to the coupled differential equations arising from the Schrodinger equation indicates the efficiency of the new approach. PMID- 11500108 TI - Applying the concept of partially ordered sets on the ranking of near-shore sediments by a battery of tests. AB - When a ranking of some objects (chemicals, geographical sites, river sections, etc.) by a multicriteria analysis is of concern, then it is often difficult to find a common scale among the criteria, and therefore even the simple sorting process is performed by applying additional constraints, just to get a ranking index. However such additional constraints, often arising from normative considerations, are controversially discussed. The theory of partially ordered sets and its graphical representation (Hasse diagrams) does not need such additional information just to sort the objects. Here, the approach of using partially ordered sets is described by applying it to a battery of tests, developed by Dutka et al. In our analysis we found the following: (1) The dimension analysis of partially ordered sets suggests that, at least in the case of the 55 analyzed samples and the evaluation by the scores, developed by Dutka et al., there is a considerable redundancy with respect to ranking. The visualization of the sediment sites can be performed within a two-dimensional grid. (2) Information, obtained from the structure of the Hasse diagram: For example six classes of sediment sites have high priority, and each class exhibits a different pattern of results. (3) Loss of information, when an aggregation of test results is used in order to guarantee complete comparability among all objects. A relation between information drawn from the graphic and the uncertainty of ranking after using an aggregation is given. (4) The sensitivity analysis identifies one test as most important, namely the test for Fecal Coliforms/Escherichia coli. This means that the ranking of samples is heavily influenced by the results of this specific test. PMID- 11500109 TI - Iterated reaction graphs: simulating complex Maillard reaction pathways. AB - This study investigates a new method of simulating a complex chemical system including feedback loops and parallel reactions. The practical purpose of this approach is to model the actual reactions that take place in the Maillard process, a set of food browning reactions, in sufficient detail to be able to predict the volatile composition of the Maillard products. The developed framework, called iterated reaction graphs, consists of two main elements: a soup of molecules and a reaction base of Maillard reactions. An iterative process loops through the reaction base, taking reactants from and feeding products back to the soup. This produces a reaction graph, with molecules as nodes and reactions as arcs. The iterated reaction graph is updated and validated by comparing output with the main products found by classical gas chromatographic/mass spectrometric analysis. To ensure a realistic output and convergence to desired volatiles only, the approach contains a number of novel elements: rate kinetics are treated as reaction probabilities; only a subset of the true chemistry is modeled; and the reactions are blocked into groups. PMID- 11500110 TI - A novel PI index and its applications to QSPR/QSAR studies. AB - A novel topological index, PI (Padmakar-Ivan index), is derived in this paper. The index is very simple to calculate and has disseminating power similar to that of the Wiener (W) and the Szeged (Sz) indices. The comprehensive studies show that the proposed PI index correlates highly with W and Sz as well as with physicochemical properties and biological activities of a large number of diversified and complex compounds. The proposed PI index promises to be a useful parameter in the QSPR/QSAR studies. The stability of each model is demonstrated by applying cross-validation test. Furthermore, more favorable comparison with other representative indices such as the Randic index is also made in order to establish the predictive ability of the PI index. The results have shown that in several cases the PI index gave better results. PMID- 11500111 TI - Monte Carlo methods for small molecule high-throughput experimentation. AB - By analogy with Monte Carlo algorithms, we propose new strategies for design and redesign of small molecule libraries in high-throughput experimentation, or combinatorial chemistry. Several Monte Carlo methods are examined, including Metropolis, three types of biased schemes, and composite moves that include swapping or parallel tempering. Among them, the biased Monte Carlo schemes exhibit particularly high efficiency in locating optimal compounds. The Monte Carlo strategies are compared to a genetic algorithm approach. Although the best compounds identified by the genetic algorithm are comparable to those from the better Monte Carlo schemes, the diversity of favorable compounds identified is reduced by roughly 60%. PMID- 11500112 TI - A linear algorithm for the Hyper-Wiener index of chemical trees. AB - An algorithm with a complexity linear in the number of vertices is proposed for the computation of the Hyper-Wiener index of chemical trees. This complexity is the best possible. Computational experience for alkanes is reported. PMID- 11500113 TI - Exhaustive generation of organic isomers. 6. Stereoisomers having isolated and spiro cycles and new extended N_tuples. AB - A new stereoisomer generation system named CAMGEC2 for generation of stereoisomers containing isolated and spiro cycles with one or more descriptors among R, S, Z, E, M, and P is developed using Graph Theory. It includes new approaches for symmetry analysis, cycle detection processes in molecular graphs in a modular way, and also an extension of the N_tuple format for linear representation of molecular graphs that keeps graph topographical information. PMID- 11500114 TI - Identification of active molecular sites using quantum-self-similarity measures. AB - A novel approach to construct theoretical QSAR models is proposed. This technique, based on the systematic use of quantum similarity measures as theoretical molecular descriptors, opens the possibility to localize and to identify the position of the bioactive part of drug molecules in situations, where the nature of the pharmacophore is not known. To test the reliability of this new approach, the method has been applied to the study of steroids binding to corticosteroid-binding human globulin. The studied molecules involved the set of 31 Cramer's steroids, often used as a benchmark set in QSAR studies. It has been shown that theoretical QSAR models based on the present procedure are superior to those derived from alternative existing approaches. In addition, a new method to measure the statistical significance of multiparameter QSAR models is also proposed. PMID- 11500115 TI - A new approach to near-infrared spectral data analysis using independent component analysis. AB - This paper presents a new approach to near-infrared spectral (NIR) data analysis that is based on independent component analysis (ICA). The main advantage of the new method is that it is able to separate the spectra of the constituent components from the spectra of their mixtures. The separation is a blind operation, since the constituent components of mixtures can be unknown. The ICA based method is therefore particularly useful in identifying the unknown components in a mixture as well as in estimating their concentrations. The approach is introduced by reference to case studies and compared to other techniques for NIR analysis including principal component regression (PCR), multiple linear regression (MLR), and partial least squares (PLS) as well as Fourier and wavelet transforms. PMID- 11500116 TI - Quest for molecular graphs with maximal energy: a computer experiment. AB - If lambda(1), lambda(2),..., lambda(n) are the eigenvalues of a graph G, then the energy of G is defined as E(G) = the absolute value of lambda(1) + the absolute value of lambda(2) +.... + the absolute value of lambda(n). If G is a molecular graph, representing a conjugated hydrocarbon, then E(G) is closely related to the respective total pi-electron energy. It is not known which molecular graph with n vertices has maximal energy. With the exception of m = n - 1 and m = n, it is not known which molecular graph with n vertices and m edges has maximal energy. To come closer to the solution of this problem, and continuing an earlier study (J. Chem. Inf. Comput. Sci. 1999, 39, 984-996, ref 7), we performed a Monte Carlo type construction of molecular (n,m)-graphs, recording those with the largest (not necessarily maximal possible) energy. The results of our search indicate that for even n the maximal-energy molecular graphs might be those possessing as many as possible six-membered cycles; for odd n such graphs seem to prefer both six- and five-membered cycles. PMID- 11500117 TI - Cage: software for a critical analysis of (2)H spin-lattice relaxation in liquid crystals. AB - A software package of Mathematica, aimed at the analysis of (2)H NMR Zeeman (T(1Z)) and quadrupolar (T(1Q)) spin-lattice relaxation times in liquid crystals in terms of diffusional models, is presented. The models most commonly used to describe internal, overall, and collective motions in liquid-crystalline phases are considered, and dynamic parameters are obtained by means of either single point or global target approaches using simulation or fitting procedures. The use of the software as a tool for highlighting the problems encountered in this kind of analysis as well as for dealing with such problems following suitable strategies is illustrated by means of applications to experimental (2)H relaxation times of three different calamitic liquid crystals. PMID- 11500118 TI - Can 3D structural parameters be predicted from 2D (topological) molecular descriptors? AB - The dihedral angle between both phenyl rings determined by photoelectron spectroscopy in a series of seven alkylbiphenyl is described by the local spectral moments of the bond matrix. This series is extended to 78 alkylbiphenyl compounds by estimating the dihedral angle from molecular mechanics force field calculations. The linear correlation obtained between this angle and the local spectral moments shown a correlation coefficient of 0.9838. This result proves that 2D (topological) descriptors can account for 3D structural parameters. A new substituent constant is calculated as the contribution of groups to the studied rotational angle by using the information encoded into the local spectral moments. This substituent constant is not linearly related to the Taft's steric constants E(S) as they have a correlation coefficient of only 0.75. These steric constants are able to account only for 71% of the variance in the studied 3D parameter. The implications for QSPR/QSAR studies of the demonstration that 2D (topological) descriptors can describe 3D structural parameters are also analyzed. PMID- 11500119 TI - QSAR for boiling points of "small" sulfides. Are the "high-quality structure property-activity regressions" the real high quality QSAR models? AB - Our investigation was motivated by a recent paper concerning "high-quality structure-property-activity regressions" (J. Chem. Inf. Comput. Sci. 2000, 40, 899-905) for boiling points of small sulfides using variable connectivity indices. We performed QSAR study of the same data set using a logically preselected solvation index and obtained a very good one-parameter regression. The structures of the whole possible set of small sulfides (C2-C6) were generated and the statistics were proven by real prediction using an external test set of sulfides. The variants of extended prediction with extrapolated data and QSAR using an expanded training set were also performed, and all these data also revealed the preference of the solvation index. A general problem of descriptive vs predictive QSAR is discussed. PMID- 11500120 TI - Helium 4 dimer in nanotubes. AB - The ground state of the helium 4 dimer is considered using the Monte Carlo technique. In a cylinder with a hard core wall, binding depends on its radius. For a small radius binding occurs as in the one-dimensional case. With an increase of the radius, the binding becomes stronger, reaches its maximum value, and then slowly diminishes. In conical geometry, that may be realized as a generalization of a cylindrical one, this dependence of the binding energy on the radius might lead to an effective force which tends to move the molecule toward the region of minimal energy. Thus, in channels, with nonhomogeneous cross sections, the particles move easier in dimer form. In addition, the square of the momentum and of the particle separation along the cylinder axis and in the plane perpendicular to it are calculated as well. PMID- 11500121 TI - 3D QSAR analyses of novel tyrosine kinase inhibitors based on pharmacophore alignment. AB - In an effort to develop a quantitative ligand-binding model for the receptor tyrosine kinases, a pharmacophore search was first used to identify structural features that are common in two novel sets of 12 molecules of the 3-substituted indolin-2-ones and 19 compounds of the benzylidene malononitriles with low-to high affinity for HER2, a kind of receptor tyrosine kinase. The common pharmacophore model based on these 31 compounds was used as a template to obtain the aligned molecular aggregate, which provided a good starting point for 3D-QSAR analysis of only the 19 benzylidene malononitriles. Two molecular field analysis (MFA) techniques, including CoMFA and CoMSIA, were used to derive the quantitative structure-activity relationships of the studied molecules. From the studied results, it was obvious that the 3D-QSAR models based on the pharmacophore alignment were superior to those based on the simple atom-by-atom fits. Considering the flexibility of the studied molecules and the difference between the active conformers and the energy-lowest conformers, the pharmacophore model can usually provide the common features for the flexible regions. Moreover, the best CoMSIA model based on the pharmacophore hypothesis gave good statistical measure from partial least-squares analysis (PLS) (q(2) = 0.71), which was slightly better than the CoMFA one. Our study demonstrated that pharmacophore modeling and CoMSIA research could be effectively combined. Results obtained from both methods helped with understanding the specific activity of some compounds and designing new specific HER2 inhibitors. PMID- 11500122 TI - Modeling boiling points of cycloalkanes by means of iterated line graph sequences. AB - A class of models for predicting boiling points of cycloalkanes is put forward, based on iterated line graphs L(i), i = 1, 2,., of the molecular graph G = L(0). Let m(i) be the number of edges of L(i), i = 0, 1, 2,. The models analyzed are of the form a(0)m(i)()(0) + a(1)m(i)(1) + a(2)m(i)(2) +. + a(k)m(ik) + b. Our optimal QSPR formulas contain m(0), m(1), m(2), m(3), and/or m(4) but never m(5) and m(6). Their precision is as good as or better than the approximations recently reported by Rucker and Rucker (J. Chem. Inf. Comput. Sci. 1999, 39, 788 802). PMID- 11500123 TI - SIRS-SS: a system for simulating IR/Raman spectra. 1. Substructure/subspectrum correlation. AB - An IR/RAMAN spectra simulation system is reported. The development of this software was based on the substructure/subspectrum relationships established for four different structural classes: small molecules, special fragments, atom centered FRELs, and bond-centered FRELs (FREL: Fragment centered on an Environment which is Limited). Four corresponding knowledge-bases (now, at a pilot stage) are constructed from usual correlation charts or data analyses of large populations of compounds using data mining techniques. PMID- 11500124 TI - A temperature-dependent quantum mechanical/neural net model for vapor pressure. AB - We present a temperature-dependent model for vapor pressure based on a feed forward neural net and descriptors calculated using AM1 semiempirical MO-theory. This model is based on a set of 7681 measurements at various temperatures performed on 2349 molecules. We employ a 10-fold cross-validation scheme that allows us to estimate errors for individual predictions. For the training set we find a standard deviation of the error s = 0.322 and a correlation coefficient (R(2)) of 0.976. The corresponding values for the validation set are s = 0.326 and R(2) = 0.976. We thoroughly investigate the temperature-dependence of our predictions to ensure that our model behaves in a physically reasonable manner. As a further test of temperature-dependence, we also examine the accuracy of our vapor pressure model in predicting the related physical properties, the boiling point, and the enthalpy of vaporization. PMID- 11500125 TI - Differential Shannon Entropy as a sensitive measure of differences in database variability of molecular descriptors. AB - A method termed Differential Shannon Entropy (DSE) is introduced to compare differences in information content and variance of molecular descriptors between compound databases. The analysis is based on histograms recording the individual and grouped distributions of molecular descriptors and calculation of Shannon entropy (SE), a formalism originally applied to digital communication. We have recently shown that SE values reflect the nonparametric variability of descriptor settings. Now the analysis has been advanced to assess differences in information content of 143 molecular descriptors in databases containing synthetic compounds, natural products, or drug-like molecules. The DSE metric captures the degree to which descriptor distributions complement or duplicate information contained in molecular databases. In our analysis, we observe significant differences for a number of descriptors and rank them according to their associated DSE values. Using DSE calculations, relative information content of different types of descriptors can be quantified, even if differences are subtle. PMID- 11500126 TI - QSPR modeling for solubility of fullerene (C(60)) in organic solvents. AB - Solubility of fullerene C(60) in 75 organic solvents was examined to develop quantitative structure-solubility relationships. Topological indices and polarizability parameter computed from refractive index were used to form the regression models. The models suggested for individual data sets such as alkanes, alkyl halides, alcohols, cycloalkanes, alkylbenzenes, and aryl halides have good predictive ability and are better than the models for the combined groups. Inclusion of an indicator parameter which is a combination of atom contributions and contributions of substituents' position in benzenes improved the predictive ability significantly. PMID- 11500127 TI - Design and development of computer-aided chemical systems: virtual labs for teaching chemical experiments in undergraduate and graduate courses. AB - An environment for the construction of virtual chemistry experiments is presented. This environment is based on the E(V) = M + m model-Experiment (Virtual) = Materials + method-proposed and described herein, which allows the representation and subsequent building of chemistry experiments in virtual 3D worlds to any degree of complexity. The object-based nature of the environment not only allows its use on the Internet but also facilitates integration with other systems, while enabling the system to represent and organize knowledge in such a way that it is available to any teaching environment dealing with chemical laboratory experiments. PMID- 11500128 TI - Use of pruned computational neural networks for processing the response of oscillating chemical reactions with a view to analyzing nonlinear multicomponent mixtures. AB - The suitability of pruned computational neural networks (CNNs) for resolving nonlinear multicomponent systems involving synergistic effects by use of oscillating chemical reaction-based methods implemented using the analyte pulse perturbation technique is demonstrated. The CNN input data used for this purpose are estimates provided by the Levenberg-Marquardt method in the form of a three parameter Gaussian curve associated with the singular profile obtained when the oscillating system is perturbed by an analyte mixture. The performance of the proposed method was assessed by applying it to the resolution of mixtures of pyrogallol and gallic acid based on their perturbating effect on a classical oscillating chemical system, viz. the Belousov-Zhabotinskyi reaction. A straightforward network topology (3:3:2, with 18 connections after pruning) allowed the resolution of mixtures of the two analytes in concentration ratios from 1:7 to 6:2 with a standard error of prediction for the testing set of 4.01 and 8.98% for pyrogallol and gallic acid, respectively. The reduced dimensions of the selected CNN architecture allowed a mathematical transformation of the input vector into the output one that can be easily implemented via software. Finally, the suitability of response surface analysis as an alternative to CNNs was also tested. The results were poor (relative errors were high), which confirms that properly selected pruned CNNs are effective tools for solving the analytical problem addressed in this work. PMID- 11500129 TI - ACD/I-Lab 4.5: an internet service review. PMID- 11500131 TI - Chromosome nondisjunction and loss induced by protons and X rays in primary human fibroblasts: role of centromeres in aneuploidy. AB - To study the origin of micronuclei induced in human primary fibroblasts by low energy protons (7.7 and 28.5 keV/microm) and X rays, we have developed a combined antikinetochore-antibody (CREST) and FISH staining with pancentromeric probes. This technique allowed us to analyze the integrity of the kinetochore and centromeric DNA structures and to assess their role in induced aneuploidy. The effect of LET on radiation-induced chromosome nondisjunction was studied in binucleated cells with centromeric-specific DNA probes for chromosomes 7 and 11. Our results indicate that, though more than 90% of radiation-induced micronuclei were CREST(-)/FISH(-), 28.5 keV/microm protons and X rays were also able to induce statistically significant increases in the number of micronuclei that were CREST(-)/FISH(+) and CREST(+)/FISH(+), respectively. One interpretation of these results could be that the protons induced chromosome loss by kinetochore detachment or by breakage in the centromeric DNA region, whereas X rays induced aneuploidy through a non-DNA damage mechanism. Nondisjunction appears to be a far more important mechanism leading to radiation-induced aneuploidy. Irrespective of the higher frequency of micronuclei induced by 28.5 keV/microm protons, the frequency of chromosome loss was markedly higher for X rays than for 28.5 keV/microm protons, strengthening the hypothesis that non-DNA targets, such as components of the mitotic spindle apparatus, may be involved in aberrations in chromosome segregation after X irradiation. PMID- 11500132 TI - Induction of a senescence-like phenotype in bovine aortic endothelial cells by ionizing radiation. AB - Treatment of confluent monolayers of bovine aortic endothelial cells (BAEC) with gamma rays resulted in the delayed appearance of cells with an enlarged surface area that were morphologically similar to senescent cells. The majority of these cells stained positively for senescence-associated beta-galactosidase (SA-beta gal), indicating that these cells are biochemically similar to senescent cells. The incidence of the senescence-like phenotype increased with dose (5-15 Gy) and time after irradiation. Cells with a senescence-like phenotype began to appear in the monolayer several days after irradiation. The onset of the appearance of this phenotype was accelerated by subculturing 24 h after irradiation. This acceleration was not entirely due to stimulation of progression through the cell cycle, since a high percentage of the senescent-like cells that appeared after subculture were not labeled with BrdUrd during the period after subculture. Prolonged up-regulation of expression of CDKN1A (also known as p21(CIP1/WAF1)) after irradiation was noted by Western blot analysis, again suggesting a similarity to natural senescence. Phenotypically altered endothelial cells were present in the irradiated monolayers as long as 20 weeks after irradiation, suggesting that a subpopulation of altered endothelial cells that might be functionally deficient could persist in the vasculature of irradiated tissue for a prolonged period after irradiation. PMID- 11500133 TI - Effect of low doses of ionizing radiation on cells cultured from the hematopoietic tissue of the Dublin Bay prawn, Nephrops norvegicus. AB - Explant cultures from the hematopoietic tissue of the Dublin Bay prawn, Nephrops norvegicus, were exposed to low doses of (60)Co gamma radiation. Cells growing from the explants were examined 7 days after irradiation using light and transmission electron microscopy and were also tested for their ability to produce signals indicative of a bystander effect. The exposed cultures displayed pronounced damage and were orders of magnitude more sensitive than the data in the literature would suggest for arthropod cells. The cultures were also more sensitive than mammalian cells that were exposed to similar doses. Cellular abnormalities included damage to cytoplasmic organelles, particularly the cytoskeleton. Abnormal mitochondria were also prominent. At low doses (0.5 Gy), nuclear damage was not apparent in the cultures, but there was evidence of a dose dependent increase in apoptosis. The irradiated cultures released a factor into the medium that was capable of inducing apoptosis and cell death in unirradiated fish and human cells. This bystander effect was of a similar magnitude to that reported for mammalian cell systems. It is suggested that these crustaceans may be highly sensitive to radiation, unlike terrestrial arthropods and certain other invertebrates, which are generally considered to be radioresistant. PMID- 11500134 TI - Production of delayed death and neoplastic transformation in CGL1 cells by radiation-induced bystander effects. AB - Other investigators have demonstrated by transfer of medium from irradiated cells and by irradiation with low-fluence alpha particles or microbeams that cells do not have to be directly exposed to ionizing radiation to be detrimentally affected, i.e. bystander effects. In this study, we demonstrate by transfer of medium from X-irradiated human CGL1 hybrid cells that the killing of bystander cells reduces the plating efficiency of the nonirradiated CGL1 cells by 33 +/- 6%. In addition, we show that the amount of cell death induced by bystander effects is not dependent on X-ray dose, and that the induction of apoptosis does not appear to be responsible for the cell death. Furthermore, we found that the reduction in plating efficiency in bystander cells is evident for over 18 days, or 22 cell population doublings, after medium transfer, despite repeated refeeding of the cell cultures. Finally, we report the novel observation that bystander effects induced by the transfer of medium from irradiated cells can induce neoplastic transformation. Exposing unirradiated CGL1 cells to medium from cells irradiated with 5 or 7 Gy increased the frequency of neoplastic transformation significantly from 6.3 x 10(-6) in unirradiated controls to 2.3 x 10(-5) (a factor of nearly four). We conclude that the bystander effect induces persistent, long-term, transmissible changes in the progeny of CGL1 cells that result in delayed death and neoplastic transformation. The data suggest that neoplastic transformation in bystander cells may play a significant role in radiation-induced neoplastic transformation at lower doses of X rays. PMID- 11500135 TI - Diverse delayed effects in human lymphoblastoid cells surviving exposure to high LET (56)Fe particles or low-LET (137)Cs gamma radiation. AB - To obtain information on the origin of radiation-induced genomic instability, we characterized a total of 166 clones that survived exposure to (56)Fe particles or (137)Cs gamma radiation, isolated approximately 36 generations after exposure, along with their respective control clones. Cytogenetic aberrations, growth alterations, responses to a second irradiation, and mutant frequencies at the Na(+)/K(+) ATPase and thymidine kinase loci were determined. A greater percentage of clones that survived exposure to (56)Fe particles exhibited instability (defined as clones showing one or more outlying characteristics) than in the case of those that survived gamma irradiation. The phenotypes of the unstable clones that survived exposure to (56)Fe particles were also qualitatively different from those of the clones that survived gamma irradiation. A greater percentage (20%) of the unstable clones that survived gamma irradiation than those that survived exposure to (56)Fe particles (4%) showed an altered response to the second irradiation, while an increase in the percentage of clones that had an outlying frequency of ouabain-resistant and thymidine kinase mutants was more evident in the clones exposed to (56)Fe particles than in those exposed to gamma rays. Growth alterations and increases in dicentric chromosomes were found only in clones with more than one alteration. These results underscore the complex nature of genomic instability and the likelihood that radiation-induced genomic instability arises from different original events. PMID- 11500136 TI - Sensitization to the cytotoxicity of melphalan by ethacrynic acid and hyperthermia in drug-sensitive and multidrug-resistant Chinese hamster ovary cells. AB - The ability of physical and pharmacological modulators to increase the cytotoxicity of melphalan was investigated in Chinese hamster ovary cells using a clonogenic cell survival assay. Hyperthermia has potential for use in cancer treatment, particularly as an adjuvant to chemotherapy or radiotherapy. Ethacrynic acid is a glutathione S-transferase inhibitor and also undergoes conjugation with glutathione. Interactions between hyperthermia (41-43 degrees C), ethacrynic acid and melphalan were evaluated in multidrug-resistant (CH(R)C5) cells with overexpression of P-glycoprotein (33.69-fold), and in drug-sensitive (AuxB1) cells. GST alpha was expressed at a higher level (3.65-fold) in CH(R)C5 cells than in sensitive cells, whereas levels of isoforms pi and mu were the same. GST pi was the most highly expressed isoform in the two cell populations. Ethacrynic acid was cytotoxic at elevated temperatures, while it caused little or no cytotoxicity at 37 degrees C. This effect occurred in drug-resistant and drug sensitive cells, and attributes thermosensitizing properties to ethacrynic acid. Ethacrynic acid (20 microM) alone did not alter the cytotoxicity of melphalan at 37 degrees C. Hyperthermia potentiated drug cytotoxicity in cells, both with and without ethacrynic acid treatment. Ethacrynic acid could be useful in cancer treatment by acting as a thermosensitizer when combined with heat and by enhancing the cytotoxicity of melphalan at elevated temperatures. A major advantage arising from the use of regional hyperthermia is the ability to target drug cytotoxicity to the tumor volume. A useful finding is that ethacrynic acid, heat and/or melphalan are also effective against multidrug-resistant cells with overexpression of P-glycoprotein. PMID- 11500137 TI - In vivo radioprotection by 5-androstenediol: stimulation of the innate immune system. AB - We showed previously that 5-androstenediol stimulates myelopoiesis, increases the numbers of circulating neutrophils and platelets, and enhances resistance to infection in gamma-irradiated mice. We have extended those studies to include monocytes, natural killer (NK) cells, eosinophils and basophils, and we have measured the activation marker CD11b using flow cytometry. Androstenediol (160 mg/kg) was administered subcutaneously to female B6D2F1 mice 24 h before whole body gamma irradiation. Androstenediol treatments increased the blood levels of neutrophils, monocytes and NK cells in unirradiated animals; decreased the numbers of circulating eosinophils; and ameliorated radiation-induced decreases in neutrophils, monocytes, NK cells, erythrocytes and platelets. The androstenediol treatments had no significant effect on the numbers of circulating B cells or T cells. CD11b labeling intensity on monocytes was decreased slightly after androstenediol treatment. In contrast, radiation or androstenediol alone caused increases in CD11b labeling intensity on NK cells. Androstenediol and radiation combined caused a marked increase in NK cell CD11b. The results indicate that androstenediol increases the numbers of the three major cell types of the innate immune system (neutrophils, monocytes and NK cells), that androstenediol-induced changes in blood elements in irradiated animals persist for at least several weeks, and that there is a significant positive interaction between radiation and administration of androstenediol in the activation of NK cells. PMID- 11500138 TI - Enhancement of tumor oxygenation and radiation response by the allosteric effector of hemoglobin, RSR13. AB - Prior studies using pO(2) microelectrodes have shown that RSR13, an allosteric modifier of hemoglobin, increases tissue oxygenation in vivo. Recently, measurements of tissue oxygenation have been performed by many investigators using blood oxygen level-dependent magnetic resonance imaging (BOLD MRI). In this study, we tested the hypothesis that the BOLD MRI signal ratio in tumors will change after administration of RSR13. NCI-H460 human lung carcinoma cells were used as a xenograft in athymic nude mice. Mice with 1-cm(3) tumors in the flank were anesthetized and mounted on the MRI apparatus, and various doses of RSR13 were administered intraperitoneally (i.p.). MR images were then acquired at 10 min intervals for up to 60 min after injection. The effect of RSR13 on tumor response was studied using the same mouse xenograft model with tumor growth delay measurements. RSR13 increased the MRI signal ratio [Intensity(t)/Intensity(t = 0)] in a dose-dependent manner, with maximum increases occurring 30 min after RSR13 was administered. An RSR13 dose of 200 mg/kg proved to be optimum. Since the MRI signal ratio has been shown previously to be linearly related to tissue oxygenation, the changes in the MRI signal ratio can be attributed to changes in tumor oxygen levels. Using a 200-mg/kg dose of RSR13, with a 10-Gy dose of radiation administered to tumors 30 min later, enhancement of radiation-induced tumor growth delay by RSR13 was observed (enhancement factor = 2.8). Thus our MRI results support and verify the previously reported RSR13-induced increase in tumor oxygenation obtained using pO(2) microelectrodes. Based upon these results and other previous studies, the mechanism of enhancement of the effect of radiation by RSR13 probably involves an increase in tumor oxygenation. PMID- 11500139 TI - Tumor cytotoxicity in vivo and radical formation in vitro depend on the shock wave-induced cavitation dose. AB - Local tumor therapy using focused ultrasonic waves may become an important treatment option. This technique exploits the ability of mechanical waves to induce thermal and nonthermal effects noninvasively. The cytotoxicity to cultured cells and biological tissues in vivo that results from exposure to ultrasonic shock waves is considered to be a nonthermal effect that is partly a consequence of ultrasound-induced cavitation. Cavitation is defined as the formation of bubbles during the negative wave cycle; their subsequent oscillation and/or violent implosion can affect surrounding structures. To investigate cavitational effects in cells and tissues, defined cavitation doses must be applied while ideally holding all other potential ultrasound parameters constant. The application of independent cavitation doses has been difficult and has yielded little knowledge about quantitative cavitation-tissue interactions. By using a special shock-wave pulse regimen and laser optical calibration in this study, we were able to control the cavitation dose independently of other physical parameters such as the pressure amplitudes, and averaged acoustic intensity. We treated Dunning prostate tumors (subline R3327-AT1) transplanted into Copenhagen rats with shock waves at three cavitation dose levels and then determined the tumor growth delay and the histopathological changes. All of the treated animals exhibited a significant tumor growth delay compared to the controls. Higher cavitation doses were associated with a greater delay in the growth of the tumor and more severe effects on tumor histopathology, such as hemorrhaging, tissue disruption, and necrosis. In vitro, the cavitation dose level correlated with the amount of radical formation. We concluded that the process of acoustic cavitation was responsible; higher cavitation doses caused greater effects in tumors both in vivo and in vitro. These findings may prove important in local tumor therapy and other applications of ultrasound such as ultrasound-mediated drug delivery. PMID- 11500141 TI - Photon W value for krypton in the M-shell transition region. AB - Absolute W values for krypton have been measured for incident X rays with energies in the range of 85 to 1000 eV, using monochromatic synchrotron radiation and a multiple-electrode ion chamber technique that yields the absolute intensity of the X-ray beam and the photoabsorption cross section. To improve the purity of the incident X rays, the electron storage ring was operated at an energy lower than the normal mode, and thin filters were used. The W values are derived from the measured photon intensity and photoabsorption cross section, using the mean charges of the residual ions obtained in previous work. A considerable oscillation of the W values with the photon energy was found in the region near the krypton 3d electron ionization edge. The results are discussed and compared with data in the literature for low-energy electrons and with the calculations from a model that includes multiple photoionization effects related to inner shell ionization. PMID- 11500140 TI - Comparisons of LET distributions measured in low-earth orbit using tissue equivalent proportional counters and the position-sensitive silicon-detector telescope (RRMD-III). AB - Determinations of the LET distribution, phi(L), of charged particles within a spacecraft in low-Earth orbit have been made. One method used a cylindrical tissue-equivalent proportional counter (TEPC), with the assumption that for each measured event, lineal energy, y, is equal to LET and thus phi(L) = phi(y). The other was based on the direct measurement of LETs for individual particles using a charged-particle telescope consisting of position-sensitive silicon detectors called RRMD-III. There were differences of up to a factor of 10 between estimates of phi(L) using the two methods on the same mission. This caused estimates of quality factor to vary by a factor of two between the two methods. PMID- 11500142 TI - The Trp53 pathway is induced in vivo by low doses of gamma radiation. AB - The induction of the Trp53 response after very low doses (0.01-1 Gy) of ionizing radiation was studied in the adult mouse using immunochemical and immunohistochemical methods. We found a detectable response at 0.01 Gy and an increased induction of Trp53 with increasing dose in both radiation-resistant and radiation-sensitive tissues. These results suggest that there is no lower threshold for induction. This response was heterogeneous, since cells that received the same dose had different staining intensities, suggesting that the induction of Trp53 is not based simply on dose-dependent responses to DNA damage. These data also demonstrate the exquisite sensitivity of the Trp53 pathway and show that this response is controlled by cell- and tissue-specific factors that have yet to be defined. PMID- 11500143 TI - Measurement of DNA damage in mammalian cells exposed in vitro to radiofrequency fields at SARs of 3-5 W/kg. AB - In the present study, we determined whether exposure of mammalian cells to 3.2 5.1 W/kg specific absorption rate (SAR) radiofrequency fields could induce DNA damage in murine C3H 10T(1/2) fibroblasts. Cell cultures were exposed to 847.74 MHz code-division multiple access (CDMA) and 835.62 frequency-division multiple access (FDMA) modulated radiations in radial transmission line (RTL) irradiators in which the temperature was regulated to 37.0 +/- 0.3 degrees C. Using the alkaline comet assay to measure DNA damage, we found no statistically significant differences in either comet moment or comet length between sham-exposed cells and those exposed for 2, 4 or 24 h to CDMA or FDMA radiations in either exponentially growing or plateau-phase cells. Further, a 4-h incubation after the 2-h exposure resulted in no significant changes in comet moment or comet length. Our results show that exposure of cultured C3H 10T(1/2) cells at 37 degrees C CDMA or FDMA at SAR values of up to 5.1 W/kg did not induce measurable DNA damage. PMID- 11500144 TI - Chernobyl thyroid tumor tissue and nucleic acid bank. PMID- 11500145 TI - Ossification of the posterior longitudinal ligament and ligamentum flavum: imaging features. AB - Ossifications of the posterior longitudinal ligament and ligamentum flavum are both special subcategories of degenerative diseases responsible for compression of the spinal cord. Ossification of the ligaments is well demonstrated by plain radiography and computed tomography. Magnetic resonance imaging noninvasively provides useful information about the degree and extent of spinal cord compression as well as the character of the ossification. T2-weighted sequences are most effective to evaluate both spinal cord compression due to the ossification and abnormal signal intensity of the spinal cord. PMID- 11500146 TI - Pustulotic arthro-osteitis: defining the radiologic spectrum of the disease. AB - Pustulotic arthro-osteitis (PAO) was termed by Sonozaki et al., who discussed the relationship between palmoplantar pustulosis (PPP) and osteoarticular inflammation. Manifestations of PAO are observed in the anterior chest wall, the spine, the pelvis, the sacroiliac joint, and the long bones. Hyperostosis is a radiological feature of PAO; furthermore, anterior chest wall involvement is common. The term SAPHO syndrome (SAPHO being an acronym for synovitis, acne, pustulosis, hypertostosis, and osteitis) has been coined to describe disease that manifests sterile inflammatory bone lesions together with skin eruptions. SAPHO syndrome groups together the following osteo-articular lesions that have been described as separate medical entities: chronic recurrent multifocal osteomyelitis (CRMO), PAO, and arthro-osteitis associated with a follicular occlusive triad. Osseous changes due to psoriasis vulgaris and generalized pustular psoriasis can be radiologically and clinically distinguishable from osseous changes due to PPP, acne, and CRMO as seronegative spondyloarthropathies. PMID- 11500147 TI - Adult T-cell leukemia. AB - Adult T-cell leukemia (ATL) is endemic in southwestern Japan, in the Caribbean islands, and in central Africa. Human T-cell lymphotropic virus type I (HTLV-I) is the etiologic agent of ATL. The clinical characteristics are (1) onset in adulthood, (2) subacute or chronic leukemia with rapidly progressive terminal course, (3) frequent skin lesions, (4) lymphadenopathy that characteristically spares the mediastinum, (5) hepatosplenomegaly, (6) hypercalcemia, and (7) a tendency toward geographical clustering. Although hypercalcemia and osteoclastic activity due to parathyroid hormone-related peptide are frequently reported histologically, radiographic abnormalities of bone are not common. Two major patterns of osteolytic lesions observed in ATL are "punched-out" lesions resembling multiple myeloma and osteolytic metastasis and subperiosteal bone resorptions similar to those in hyperparathyroidism. PMID- 11500148 TI - Phalangeal microgeodic syndrome. AB - Phalangeal microgeodic syndrome specifically affects the phalanges in infants. The radiographic findings are characterized by small lacunae measuring less than 1 mm in diameter in the phalanges. The symptoms usually subside spontaneously within 2 to 3 months with no residual deformity. Accurate diagnosis is possible by radiologists with knowledge on this disease entity. PMID- 11500149 TI - Elastofibroma dorsi: CT and MR imaging findings. AB - Elastofibroma dorsi is a benign fibroelastic tumor of unknown etiology that arises bilaterally or unilaterally at the lower subscapular level. It can be easily diagnosed on computed tomography or magnetic resonance images and should not be mistaken for a malignant tumor. PMID- 11500150 TI - Osteoarticular tuberculosis. AB - The radiological features of osteoarticular tuberculosis are quite variable. The rareness of AIDS-associated tuberculosis and relatively mild course and imaging features may be the common feature of osteoarticular tuberculosis in Japan. Imaging features are generally similar to other low-grade infections, and liberal (often incomplete) use of antibiotics for the treatment of pyogenic infection may also make the differential diagnosis difficult on many occasions. PMID- 11500151 TI - Dialysis-related amyloidosis. AB - Dialysis-related amyloidosis (DRA) is a well-recognized complication in patients on long-term hemodialysis. It occurs secondary to the deposition of beta(2) microglobulin, preferentially in the musculoskeletal tissues. Plain radiography demonstrates advanced DRA findings such as bone erosions and cystic lesions, but it is not suitable for the demonstration of earlier changes. Magnetic resonance imaging and ultrasound are sufficient for the detection of amyloid deposition in the periarticular soft tissues and in the evaluation of spinal complications. PMID- 11500152 TI - Osteoporosis in the Japanese population. AB - The characteristic of osteoporosis in the Japanese population is different from that in the Caucasian population. The different incidence of vertebral and hip fracture may be related to both genetic and lifestyle factors. In the aspect of genetics, approximately 77% of the Japanese have the bb genotype of the vitamin D receptor gene, which is considered to be associated with higher bone mass and lower bone turnover than the Bb genotype. The incidence of vertebral fracture is significantly lower in the younger birth cohorts in both genders. However, the total number of hip fractures rapidly grows as the elderly population increases in Japan. It may be explained by some epidemiological studies that the traditional Japanese lifestyle may prevent hip fractures in the Japanese elderly. The knowledge of the differences of osteoporosis between Caucasian and Japanese people must contribute to the prevention of osteoporosis and its complications. PMID- 11500153 TI - Values and problems in MR imaging for the evaluation of low back pain (orthopedic surgeon's view). AB - MR imaging has a significant diagnostic value, but abnormal findings revealed by MR imaging do not always cause symptoms. MR imaging findings should be confirmed by subjective symptoms, objective findings, and functional diagnostic procedures. PMID- 11500154 TI - Proper use of MR imaging for evaluation of low back pain (radiologist' view). AB - Magnetic resonance (MR) imaging has been recognized as a modality of choice in the evaluation of the spine. However, morphological abnormalities demonstrated by MR imaging do not always reflect low back pain (LBP). MR imaging should be interpreted with consideration of full clinical signs, symptoms, and other relevant background. At the same time, MR findings potentially relevant to LBP, such as morphological abnormalities of the disc, presence or absence of high intensity zone, abnormalities of the end plate and adjacent marrow signal, degree of nerve root compression, and degeneration of the facet joint, should be described without omission. PMID- 11500155 TI - Evaluation of patients with low back pain: a need for a standardized approach (radiologist's view). AB - Magnetic resonance (MR) imaging has been recognized as a modality of choice in the evaluation of the spine. However, morphological abnormalities demonstrated by MR imaging do not always reflect low back pain (LBP). MR imaging should be interpreted with consideration of full clinical signs, symptoms, and other relevant background. At the same time, MR findings potentially relevant to LBP, such as morphological abnormalities of the disc, presence or absence of high intensity zone, abnormalities of the end plate and adjacent marrow signal, degree of nerve root compression, and degeneration of the facet joint, should be described without omission. PMID- 11500156 TI - Application of MR imaging for internal derangement of the knee (orthopedic surgeon' view). AB - Magnetic resonance imaging is a noninvasive imaging modality with clear contrast and superior spatial resolution. These characteristics are especially useful for detecting pathology of the soft tissue of the knee joint, such as the menisci, ligaments and articular cartilage, which are difficult to diagnose using plain X ray examination. MRI has become one of the first choice diagnostic modalities for the internal derangement of the knee joint, and is generally replacing some invasive arthrographic or arthroscopic examination. Pathology of the articular cartilage cannot yet be depicted clearly by MRI. We expect further development of the spatial resolution of MRI to make possible the detection of chondral lesions more clearly and precisely in the near future. PMID- 11500157 TI - Proper use of MR imaging in internal derangement of the knee (orthopedic surgeon's view). AB - MR imaging of the knee is a highly accurate examination in terms of the detection of meniscal tears and cruciate ligament ruptures among internal derangement of the knee. The overall accuracy in more than 2000 published cases reached nearly as high as 90% in meniscal tears, and 94% in anterior cruciate ligament (ACL) and 99% in posterior cruciate ligament (PCL) ruptures. Furthermore, the negative predictive value of the knee MR imaging, the number of true negative examination results divided by the numbers of true negative plus false-negative examination results, reaches almost 100% because of its very low false-negative rate. Therefore, from the orthopedic surgeon's point of view, even though the medical cost becomes a great concern, MR imaging has prime importance in the diagnosis of the internal derangement of the knee. PMID- 11500158 TI - MR imaging of cartilage lesions of the knee: what is the clinical indication? (radiologist's view). AB - This commentary focuses on the clinical application of magnetic resonance (MR) imaging in the assessment of articular cartilage of the knee. MR imaging is superior to conventional imaging techniques for evaluation of the articular cartilage. Several pulse sequences are advocated for demonstration of the normal articular cartilage and cartilage lesions. However, a consensus has not yet been reached in regard to the most appropriate sequence for evaluation of the articular cartilage. Early morphological changes in the degenerative cartilage are not reliably diagnosed even with the use of tailored MR imaging techniques. The detection of the biochemical changes of cartilage will serve as an important tool for the early diagnosis of cartilage degeneration in the near future. Further prospective studies are needed to establish the role of MR imaging in clinical use. PMID- 11500159 TI - Current practice of knee MR imaging in Japan (radiologist's view). AB - The knee is probably the most frequent site for MR imaging of the extremities in most institutions, and it has significantly changed the practice of knee surgery, including arthroscopy. The following commentary by a radiologist discusses the growing number of MR units, which facilitates increases in the number of MR examinations of the knee in Japan. This easy access to MRI includes elderly patients with degenerative knee disorders, as well as younger patients with traumatic injuries. With prolonged life expectancy, indication for MRI of the knee will be expanded, despite its high cost. Precise evaluation of articular cartilage in the early stages of regeneration is still a challenge; proper use of MR imaging for this disorder must be considered. PMID- 11500160 TI - MR imaging for evaluation of early rheumatoid arthritis. AB - In rheumatoid arthritis (RA), due to its superior contrast resolution and tomographic nature, magnetic resonance (MR) imaging can depict soft tissue and joint involvement better than plain radiography. Active synovitis and pannus are shown by a wide variety of contrast on T1- and T2-weighted images. They are markedly enhanced by intravenous gadolinium-chelate injection. Fat-suppressed T1 weighted imaging with gadolinium enhancement is the most sensitive technique to demonstrate these tissues. Compared with plain radiography, MR imaging is more sensitive and equally specific in the diagnosis of early RA compared with plain radiography. MR imaging is useful in the diagnosis and treatment of patients who are suspected of having early RA. PMID- 11500161 TI - Muscular sarcoidosis. AB - There are four types of muscular sarcoidosis: nodular, chronic myopathy, acute myositis, and asymptomatic. The nodular type is important because it may be confused with a soft tissue tumor. Magnetic resonance (MR) imaging provides specific findings. A star-shaped central structure of decreased signal intensity, a "dark star" sign, is seen on the axial images. A long nodule with an inner stripe of decreased signal intensity and outer stripes of increased signal intensity, a "three stripes" sign, is seen on the coronal and sagittal images. In the chronic myopathy type, the role of MR imaging is limited. Gallium scintigraphy, which reflects activity of inflammation, may show increased uptake. In the acute myositis type, MR imaging shows increased signal intensity, and gallium scintigraphy shows increased uptake; however, these findings are nonspecific. PMID- 11500162 TI - Intertrabecular vertebral metastases: metastases only detectable on MR imaging. AB - Metastatic intertrabecular vertebral tumors that infiltrate the marrow space without trabecular bone alteration are not visible on radiographs or bone scans. To understand the clinical importance of intertrabecular metastases, their histological and radiological aspects were reviewed based on an examination using 69 cadavers. Metastatic tumors were found in 940 of 1653 vertebral bodies. Radiography of the specimen demonstrated lesions in 485 of 940 vertebral bodies (51.6%). Bone scintigraphy showed lesions in 109 of 415 vertebral bodies (26.3%) with tumors examined within 3 months before autopsy, whereas magnetic resonance (MR) images detected 132 of 146 (90.4%) lesions. The intertrabecular metastases were found in 36.9% of the metastatic lesions and was difficult to see on radiography (5.8%) and bone scans (3.3%) whereas MR images detected most of them (94.6%). The intertrabecular metastasis is the most common type of skeletal metastases and is only detectable on MR images. PMID- 11500163 TI - Nuclear imaging of bone tumors: thallium-201 scintigraphy. AB - (201)Tl scintigraphy provides important information for the management of patients with bone tumors. Although the role for staging the disease of bone tumors and for differentiation of benign from malignant lesions is limited, (201)Tl scintigraphy reflects the disease activity after treatment and it should be used to determine the treatment response and for early diagnosis of recurrence. Baseline study is essential for future reference to evaluate the response to preoperative chemotherapy and to detect recurrence after surgery. Sequential (201)Tl scintigraphy before and after treatment is useful in assessing the grade of response of the tumor to chemotherapy. The early prediction of chemotherapeutic effect by (201)Tl scintigraphy during treatment will affect the management of patients who do not respond to the therapy. This is of special importance to determine whether the patient needs an amputation or a limb-salvage surgery. PMID- 11500164 TI - Nuclear imaging of bone tumors: FDG-PET. AB - Positron emission tomography (PET) has become a very useful adjunct to anatomic imaging techniques, because it can provide an in vivo method for quantifying functional metabolism in normal and diseased tissues. Clinical trials with [(18)F] 2-deoxy-2-fluoro-D-glucose (FDG), the most commonly used radiolabeled tracer for PET imaging, has demonstrated increased accumulation of FDG in cancer tissue. FDG-PET is now widely used for the detection, differentiation, grading, staging, and monitoring of various neoplasms. However, the significance of FDG PET in such evaluations of primary bone tumors and tumor-like lesions has not been extensively elucidated. In this article, we present recent advances in FDG PET studies for evaluating primary bone tumors and tumor-like lesions. PMID- 11500165 TI - Total en bloc spondylectomy. AB - We have developed a surgical technique of total en bloc spondylectomy (TES) through a posterior approach applied to solitary metastatic vertebral tumors. Our TES is a radical en bloc resection of a whole vertebra in two parts with an oncological wide margin to obtain local curability. It consists of two steps: (1) en bloc laminectomy and setting of posterior spinal instrumentation for stabilization and (2) en bloc corpectomy and replacement by a vertebral prosthesis. We have performed this procedure in 72 patients with malignant vertebral tumors. Three of these 72 patients had local recurrence. TES is the most radical method for malignant spinal tumors. PMID- 11500166 TI - Pasteurized autogenous bone graft for reconstruction after resection of malignant bone and soft tissue tumors: imaging features. AB - Our experimental study revealed that pasteurization was sufficient for killing all tumor cells in any massive bone and that the osteoinductivity and mechanical strength of pasteurized bone were about same as that of a fresh bone graft and better than boiled or autoclaved bone. We have performed reconstruction surgery using pasteurized autogenous bone graft since 1990. Resected bone was warmed for 30 minutes in a homeothermal heater set at 60 degrees C and then placed back to the original anatomic site. The graft was fixed with a plate or intramedullary nail with or without bone cement. Based on our experience in 31 patients, no local recurrence was seen in the grafts. Among 24 patients who can be adequately evaluated, incorporation of graft was complete in 17 cases and partial in 4 cases. In 18 of 21 patients, uptake of radionuclide in the grafted pasteurized bone was detected on bone scintigraphy approximately 6 months after surgery, and the uptake increased gradually, which suggested revascularization and remodeling of grafted bone. PMID- 11500167 TI - Sagittal splitting laminoplasty for spinal canal enlargement for ossification of the spinal ligaments (OPLL and OLF). AB - Patients have been followed up radiologically after sagittal splitting laminoplasty (SSL) for ossification of the posterior longitudinal ligament (OPLL) and ligamentum flavum. The enlarged spinal canal was maintained satisfactorily in all the cases, and the posterior spinal elements were also almost completely restored. In most cases, the range of the vertebral motion was good with more than 50% of the preoperative status. Postoperative instability of the vertebral column was not identified in any case. Mild cervical kyphosis was noted in 6% of cervical OPLL patients. There were no patients with neurological deterioration. Using SSL, reconstruction and restoration of the posterior spinal elements and retention of the enlarged spinal canal were accomplished safely and adequately. PMID- 11500168 TI - Hyperglycemia-induced ascorbic acid deficiency promotes endothelial dysfunction and the development of atherosclerosis. AB - Dehydroascorbic acid, the oxidized form of vitamin C, is transported into mammalian cells via facilitative glucose transporters and hyperglycemia inhibits this process by competitive inhibition. This inhibited transport may promote oxidative stress and contribute to the increase in atherosclerotic cardiovascular disease observed in patients with diabetes mellitus. This review explores the importance of this proposed mechanism in light of current research. For example, recent reports suggest that administration of antioxidants, such as vitamin C, may slow atherogenesis by improving endothelium-dependent vasodilation in individuals with abnormal glucose and lipid metabolism, perhaps by preventing the oxidation of nitric oxide, an important regulator of vasomotor tone. Endothelial dysfunction plays a key role in the development of atherosclerosis and endothelial cells may be particularly affected by hyperglycemia-induced ascorbic acid deficiency as they line the interior of blood vessels. In addition, we discuss evidence of several other mechanisms by which vitamin C status may affect the development of atherosclerotic cardiovascular disease, particularly its inverse relationship to multiple cardiovascular disease risk factors and indicators. Given these factors, vitamin C administration is recommended during periods of both acute and chronic hyperglycemia to help preserve endothelial function. PMID- 11500169 TI - Chlamydia pneumoniae infection accelerates hyperlipidemia induced atherosclerotic lesion development in C57BL/6J mice. AB - Considerable evidence of an association between Chlamydia pneumoniae infections and cardiovascular disease has emerged. Animal models using genetically altered mice and hypercholesterolemic rabbits have shown a pathogenic role of C. pneumoniae in accelerating atherosclerotic plaque development. In the present study, we evaluated the effect of chronic C. pneumoniae infection on atherosclerosis in C57BL/6J mice, fed either a regular chow diet or a high fat, high cholesterol diet. Infected animals on an atherogenic diet developed significantly larger lesion areas compared with control mice at 18 weeks (2.5 fold increase; 4177+/-777 vs. 1650+/-808 microm(2); P<0.05) and 24 weeks of age (3.3-fold increase; 14139+/-4147 vs. 4298+/-869 microm(2); P<0.02). This study shows that chronic C. pneumoniae infection accelerates atherosclerotic lesion development in diet induced hypercholesterolemic mice, indicating that C. pneumoniae is a co-risk factor of hyperlipidemia in atherogenesis. PMID- 11500170 TI - Antiplatelet agent cilostazol potentiates adipocyte differentiation of 3T3-L1 cells. AB - Cilostazol is an antiplatelet drug, which has beneficial effects in treatment of intermittent claudication and decreases serum triacyiglycerol level in these patients. In this study, we examined adipogenic potency of cilostazol using 3T3 L1 preadipocyte cell line because cilostazol is one of the tissue specific phosphodiesterase (PDE) inhibitors. Addition of cilostazol into the differentiation medium including insulin and dexamethasone, induced the adipocyte differentiation without isobutyl methylxanthine (IBMX). Compared with the cells incubated with vehicle, the cells treated with cilostazol contain much more lipid droplets in the cells 6 days after induction of differentiation. Adipocyte specific gene like stearoyl-CoA desaturase was strongly induced after addition of cilostazol. C/EBPbeta, which is induced by IBMX was also induced by cilostazol. These findings suggest a possibility that adipogenic effect of cilostazol is one of the mechanisms, by which this agent decreases blood triacylglycerol level in the intermittent claudication patients. PMID- 11500171 TI - Induction of apoptosis and inhibition of migration of inflammatory and vascular wall cells by cerivastatin. AB - Statins are thought to play a role in directly affecting immune and mesenchymal cells. Since cerivastatin's pleiotropic effects are poorly investigated, we were interested to find out whether this drug can modulate leukocyte and vessel wall cell functions. Leukocyte migration was tested in modified Boyden microchemotaxis chambers and oxygen radical production was measured fluorometrically. Transendothelial migration experiments were performed with human umbilical vein endothelial cells and neutrophils. Neutrophil, monocyte, and vascular smooth muscle cell caspase-3 activity and annexin-V binding were quantified by FIENA and FACS, respectively. Cerivastatin [10 pM to 100 microM] decreased leukocyte chemotaxis towards interleukin-8 or RANTES. Migration of cells was completely restored by addition of mevalonic acid. In neutrophils, cerivastatin [100 microM] reduced transendothelial migration, whereas treatment of endothelial cells failed to affect transmigration. Neutrophil respiratory burst activity was unaffected by cerivastatin. At concentrations of 10 nM or higher, cerivastatin increased the rate of apoptosis in phagocytes and smooth muscle cells. Results show that cerivastatin is able to inhibit leukocyte chemotaxis, and that cerivastatin induces neutrophil, monocyte, and smooth muscle cell apoptosis. The drug's impact on transendothelial migration is due to its effects on neutrophils. In addition to its lipid-lowering effects, pharmacological properties of cerivastatin may include modulatory actions in leukocytes and mesenchymal cells. PMID- 11500172 TI - A multiplex PCR-based DNA assay for the detection of paraoxonase gene cluster polymorphisms. AB - Paraoxonase (PON) is a high-density lipoprotein (HDL) associated protein which is supposed to protect low-density lipoprotein (LDL) against oxidation and to play a role in the development of atherosclerosis. Interindividual variability in serum PON activity is attributable to common variants in components of the PON gene cluster on chromosome 7. We describe experimental conditions that permit the simultaneous determination of three common PON polymorphisms (PON1-192, PON1-55 and PON2-311) that are tightly associated with an increased risk of atherosclerosis. We used a multiplex PCR-based DNA assay using mismatch primers that introduce a unique recognition site for the endonuclease HinfI in the PCR products in case of presence of the R allele of PON 1-192, of the L allele of PON1-55 and of the S allele of PON2-311. The restriction analysis with HinfI allows to identify an electrophoretic band pattern which is specific for the combination of the three polymorphisms. This technique could be applied in the association studies aimed at assessing the role of PON and their polymorphisms in many clinical settings. In a preliminary study on a small population sample from south Italy about 10% of chromosomes exhibited the presumed risk-related haplotype R(192)/L(55)/S(311). PMID- 11500173 TI - C-terminal fragment of alpha1-antitrypsin activates human monocytes to a pro inflammatory state through interactions with the CD36 scavenger receptor and LDL receptor. AB - Monocyte scavenger receptor, CD36 has been implicated in the pathogenesis of atherosclerosis as a major oxidised LDL receptor mediating lipid accumulation and foam cell formation. Previously, we found that treatment of monocyte cultures with the carboxyl terminal fragment of alpha1-antitrypsin (C-36) increases lipid binding and uptake, induces LDL receptor mRNA and CD36 receptor protein expression, and also significantly increases production of pro-inflammatory molecules. To assess the role of the CD36 receptor in proatherogenic monocyte activation by the C-36 fragment, we tested whether specific anti-CD36 receptor antibodies would block the effects of C-36 on monocyte activation. We find that pre-incubation of cells with anti-LDL and anti-CD36 receptor antibodies (10 microg/ml) blocks binding of 125I-C-36 by about 50%. Similarly, cells pre incubated with oxidised LDL or native LDL at concentrations from 2.5 to 10 microg/ml showed a loss of 125I-C-36 binding (up to 49 and 57%) and uptake (up to 47 and 59.8%), respectively. In parallel experiments, monocytes were first incubated for 1 or 6 h with anti-CD36 antibodies (10 microg/ml) prior to adding C 36 peptide. Anti-CD36 antibodies suppressed C-36-induced production of gelatinase B, monocyte chemoattractant protein-1, interleukin-6 and cellular oxygen consumption to control levels, whereas levels of TNFalpha were unaffected. In contrast, saturation of LDL receptors with excess of anti-LDL (20 microg/ml) significantly inhibited C-36 induced TNFalpha levels. Results indicate that the C 36 peptide binds to both LDL and CD36 scavenger receptors which involves selective upregulation of pro-inflammatory molecules and activation of the respiratory burst in human monocytes. This also supports important roles for CD36 and LDL receptors in atherogenesis and suggests that blockade of CD36 receptor can be protective in pro-inflammatory activation of human monocytes. PMID- 11500174 TI - Comparison of endothelial function evaluated by strain gauge plethysmography and brachial artery ultrasound. AB - Strain gauge plethysmography and brachial artery ultrasound are widely used to study endothelial function. No data on correlation between these two procedures are reported. The present study compared these two methods and investigated the correlation between vasodilation and brachial wall shear stress. In six healthy subjects and ten patients with hypertension or obesity, strain gauge plethysmography was performed in resting conditions and after infusion of 7.5,15 and 30 microg/min of acetylcholine, and brachial artery ultrasound in resting conditions and after 5 min hand ischemia. Wall shear stress was calculated as: blood viscosity x blood velocity/internal diameter. Forearm blood flow following acetylcholine infusion increased more in healthy subjects than in patients with hypertension or obesity. In addition, brachial artery dilated more in the former group. Change in brachial artery diameter correlated with change in forearm blood flow, calculated as area under the curve of acetylcholine infusion (r=0.739, P<0.001). Wall shear stress was higher in healthy subjects (67.8+/-20.0 dynes/cm(2)) than in patients with either hypertension or obesity (39.2+/-16.7, P<0.001), and correlated with variations of diameter (r=0.796, P<0.0002), and marginally of blood flow (r=0.516, P<0.05). The present findings demonstrate that there is a high correlation between endothelial function evaluated by strain gauge plethysmography and brachial artery ultrasound. Wall shear stress correlates with brachial artery diameter change following hand ischemia, and marginally with blood flow change following acetylcholine infusion. PMID- 11500175 TI - 25-hydroxycholesterol induces lipopolysaccharide-tolerance and decreases a lipopolysaccharide-induced TNF-alpha secretion in macrophages. AB - Several different oxysterols are formed when LDL is oxidized. The role of oxysterols in the inflammatory process in the atherosclerotic plaque is not totally elucidated. In this study we have investigated the effect of four different oxysterols on an LPS-induced TNF-alpha secretion in human macrophages. Cultured human macrophages were incubated with 7-keto-, 7beta-hydroxy-, 27 hydroxy- and 25-hydroxycholesterol for 24 h before exposure to endotoxin (LPS) for 3 h. All oxysterols, except 7-ketocholesterol, significantly decreased an LPS induced TNF-alpha secretion. The most pronounced effect was obtained with 25 hydroxycholesterol, where the TNF-alpha secretion was reduced to 8%. This decreased effect was also found on the TNF-alpha mRNA level. The decreased LPS induced TNF-alpha secretion coincided with an increased binding of the transcription factors Sp1 and Sp3 to the TNF-alpha promoter. In vitro studies of the TNF-alpha promoter suggests possible interactions between Sp1 and Sp3 and the NF-kappaB transcription factor complex that might affect the transcriptional initiation. PMID- 11500176 TI - Demonstration of Chlamydia pneumoniae in atherosclerotic arteries from various vascular regions. AB - Chlamydia pneumoniae (CP) has been reported to be a pathogenic agent in the mechanism leading to atherosclerosis. The majority of available data is focused mainly on coronary artery disease whereas the distribution of CP in different areas, associated with atherosclerotic disorders, has not been completely clarified. In this study we investigated the presence of CP in atheromasic plaques from five different vascular areas (basilary artery, coronary artery, thoracic aorta, abdominal aorta, renal arteries) using nested polymerase chain reaction (PCR) and immunohistochemical staining (IHC), in order to establish the putative association of CP with atherosclerotic disease. The same atheromasic plaques were also tested for the presence of Helicobacter pylori (HP) and cytomegalovirus (CMV), other putative agents of atherosclerosis, using a nested PCR technique. Our data indicate that the presence of CP can be demonstrated in 100% of patients tested, considering globally the five areas of analysis. On the other hand the presence of HP has been demonstrated in four out of 18 patients (22.2%), and CMV only in three out of 18 (16.6%). Our results strongly suggest an association between CP and atherosclerosis and highlight the need for the detection of CP in multiple vascular areas of the same patient. PMID- 11500177 TI - The rise of the plasma lipid concentration elicited by dietary sodium chloride restriction in Wistar rats is due to an impairment of the plasma triacylglycerol removal rate. AB - Studies in humans have indicated that dietary salt restriction raises plasma levels of total cholesterol (TC) and triacylglycerols (TAG). In order to explain the mechanisms involved, a rat experimental model was developed consisting of chronic feeding ad libitum isocaloric diets with variable sodium chloride contents. Rates of synthesis of plasma TAG were measured either as the increase of plasma TAG after blocking its removal from plasma by the intra-arterial pulse infusion of Triton-WR 1339, or as the plasma rate of incorporation of [(14)C] oleic acid [(14)C]-TAG. Plasma TAG removal rate was determined by the intra arterial pulse infusion of a lipid emulsion. Severe salt restriction increased the plasma concentrations of TAG (71%) and of TC (10%). This result was not due to modification of the rate of synthesis of plasma TAG but was attributed to a 55% slower rate of removal of the TAG-containing lipoproteins. An increased plasma non-esterified fatty acid concentration, probably due to a salt restriction-related insulin resistance, may have impaired the activity of the enzyme lipoprotein lipase. PMID- 11500178 TI - Freunds adjuvant alone is antiatherogenic in apoE-deficient mice and specific immunization against TNFalpha confers no additional benefit. AB - TNFalpha participates in the pathogenesis of atherosclerosis. The effect of immunization against TNFalpha on development of advanced vascular lesions in atherosclerosis-susceptible apoE-deficient mice was investigated. At 5-7 weeks of age, animals received immunization with either Freunds adjuvant and a recombinant antigenic TNFalpha molecule (TNF106), Freunds adjuvant alone, or no immunization. All mice received a Western-type high-fat diet for 12 weeks. Aortic sinus lesion area was determined by microscopic morphometry, the total aortic arch cholesterol content was determined by gas chromatography, and antibodies against TNFalpha, malondialdehyde-modified low density lipoprotein, or heat shock protein 60, were assessed by ELISAs. Immunization with TNF106 induced high-titered circulating antibodies against TNFalpha (n=23), and these antibodies were not detected in mice immunized with Freunds adjuvant alone (n=22), or in non-immunized control animals (n=25). After 12 weeks, the atherosclerotic lesion size was significantly reduced in immunized animals, whether they had been immunized with TNF106 or Freunds adjuvant alone, and the total lesional cholesterol content was decreased in mice immunized with TNF106. There were no correlations between circulating antibody titers and plaque size, total aortic arch cholesterol content, or plasma lipid levels, respectively. Administration of Freunds adjuvant alone can thus reduce formation of mature atherosclerotic lesions in apoE-deficient mice and this response is not modified by specific immunization against TNFalpha. PMID- 11500179 TI - 2,4-Decadienal downregulates TNF-alpha gene expression in THP-1 human macrophages. AB - Oxidized lipoproteins inhibit TNF-alpha secretion by human THP-1 macrophages due, at least in part, to aldehydes derived from the oxidation of polyunsaturated fatty acids. This study extends these findings by investigating the effect of three aldehydes (2,4-decadienal (2,4-DDE), hexanal and 4-hydroxynonenal (4-HNE)) on TNF-alpha and IL-1beta mRNA expression. The 2,4-DDE and 4-HNE showed considerable biological activity which induced cytotoxicity on THP-1 macrophages at concentration of 50 microM. Hexanal, on the other hand, had a lower cytotoxic capacity and concentration of 1000 microM was needed for the effect to be observed. Exposure of THP-1 macrophages to aldehydes for 24 h inhibited TNF-alpha mRNA expression but increased or did not affect IL-1beta mRNA levels. The inhibitory action of 2,4-DDE was dose dependent and began at 5 microM (46%, P<0.001). The effect of 4-HNE was less inhibitory than 4-DDE but only when cytotoxic concentrations were used (50 microM). Very high concentrations of hexanal (200 microM) were needed to inhibit TNF-alpha expression (23%, P<0.001). This downregulation of TNF-alpha gene expression by 2,4-DDE was parallel to a lower protein production. These data indicate that low levels of 2,4-DDE may modulate inflammatory action by inhibiting TNF-alpha mRNA gene expression and that the biological activity of 2,4-DDE may be involved in the development of atherosclerosis. PMID- 11500180 TI - Intravascular hemolysis increases atherogenicity of diet-induced hypercholesterolemia in rabbits in spite of heme oxygenase-1 gene and protein induction. AB - Free radical mediated oxidation of apoB lipoproteins in the arterial intima appears to contribute to atherogenicity of the entrapped particles. A plausible pathogenic mechanism for oxidation is the one induced by heme leaking from erythrocytes that is then carried into the arterial wall by its high affinity for lipoproteins. In the intima, in the presence of H(2)O(2) secreted by macrophages, heme can be a potent oxidant. To study the role of heme as a promoter of oxidative stress damage in vivo we used a model of intravascular hemolysis (IVH) caused by phenylhydrazine in rabbits with and without diet-induced moderate hypercholesterolemia (MHC). Evaluation of the antioxidant status of plasma indicated that at the end of the treatment period this was compromised by the MHC IVH. After 10 weeks the animals with combined MHC-IVH showed more of the aorta surface covered by lesions (27%+/-8, mean (SD) than the animals with only MHC (11%+/-7), in spite of having similar plasma levels of VLDL+LDL lipoproteins. The animals with only IVH, as well as the controls, showed minimal lesions (<1%). Heme oxygenase (HO-1) expression in aorta and other tissues was markedly increased in the group with MHC-IVH and it was correlated with the extent of IVH. The data suggest that the oxidative stress associated with IVH potentiates the atherogenicity of moderate hypercholesterolemia and that in spite of a strong induction of HO-1 this is not sufficient to counteract the atherogenicity of the combined condition. PMID- 11500181 TI - Thiazolidinediones, peroxisome proliferator-activated receptor gamma agonists, regulate endothelial cell growth and secretion of vasoactive peptides. AB - Insulin resistance has been highlighted as a common causal factor for glucose intolerance, hypertension and dyslipidemia, all of which are cardiovascular risk factors. A new class of antidiabetic agents, thiazolidinediones (TZDs), has been developed and demonstrated to improve insulin sensitivity. TZDs are high affinity ligands for peroxisome proliferator-activated receptor gamma (PPARgamma), the crucial transcription factor for adipocytes. Recent studies showed that PPARgamma is also expressed in monocytes/macrophages and is suggested to be involved in atherosclerosis. We could detect PPARgamma gene transcript in several cultured endothelial cells (human aortic endothelial cells (HAoECs), human coronary artery endothelial cells (HCAECs), human umbilical vein endothelial cells (HUVECs) and bovine carotid artery endothelial cells (BAECs)) as well as human coronary arteries we examined. Since endothelial dysfunction is critical for atherosclerosis, we investigated the effects of TZDs, troglitazone (TRO) and pioglitazone (PIO), on endothelial cell growth and secretion of C-type natriuretic peptide (CNP), which we demonstrated as a novel endothelium-derived relaxing peptide, and endothelin (ET), a potent vasoconstrictor, using HAoECs, HCAECs, HUVECs and BAECs. When all these cultured endothelial cells were daily treated with TRO and PIO for 5 days, both TRO and PIO (10(-8)M) significantly stimulated (3)H-thymidine incorporation of all these endothelial cells. In contrast, higher dose of TRO and PIO (10(-5)M) significantly suppressed DNA synthesis. TRO and PIO also exerted the compatible effect on the increase of cell numbers. TRO and PIO significantly enhanced CNP secretion from BAECs. In contrast, ET secretion from BAECs was suppressed by both TRO and PIO in a dose dependent manner. The results of the present study suggest that TZDs modulate endothelial functions, including regulation of endothelial cell growth and secretion of endothelium-derived vasoactive substances, which affect vascular tone and remodeling in the process of atherosclerosis. PMID- 11500182 TI - Cilostazol represses vascular cell adhesion molecule-1 gene transcription via inhibiting NF-kappaB binding to its recognition sequence. AB - Cilostazol is a specific inhibitor of cAMP phosphodiesterase, which is used for treatment of ischemic symptoms of peripheral vascular disease. Although cilostazol has antiplatelet and vasodilator properties, its effect on the expression of adhesion molecules in vascular endothelium is not known. In the present investigation, we examined the effect of cilostazol on the expression of vascular cell adhesion molecule-1 (VCAM-1) in cultured vascular endothelial cells. Cilostazol strongly inhibited tumor necrosis factor (TNF)-alpha-induced expression of VCAM-1 protein and its mRNA. In addition, cilostazol reduced TNF alpha-induced U937 cell adhesion to the vascular endothelial cells. In transient transfection studies, cilostazol inhibited TNF-alpha-induced transcriptional activation of VCAM-1 promoter. Electrophoretic mobility shift assays revealed that cilostazol repressed TNF-alpha-induced increase in binding of the transcription nuclear factor-kappaB (NF-kappaB) to its recognition site of VCAM-1 promoter. Cilostazol, however, failed to prevent nuclear translocation of the NF kappaB p65 protein. These data indicate that cilostazol repressed VCAM-1 gene transcription in cultured vascular endothelial cells, via inhibiting NF-kappaB binding to its recognition sequence. Since the expression of the adhesion molecule is one of the earliest events occurred in atherogenic process, cilostazol might have the potential to prevent atherosclerosis at least via inhibition of the expression of the adhesion molecule. PMID- 11500183 TI - Increasing levels of dietary homocystine with carotid endarterectomy produced proportionate increases in plasma homocysteine and intimal hyperplasia. AB - PURPOSE: The role that homocysteine may play in post-carotid endarterectomy (CEA) restenosis due to intimal hyperplasia is not well understood. This study was designed to investigate the effects of different levels of dietary homocystine on: (1) plasma homocysteine; (2) post-CEA intimal hyperplasia; and (3) levels of the methyl donor S-adenosylmethionine (SAM) and its counterpart S adenosylhomocysteine (SAH) in the homocysteine pathway. METHODS: Male rats were fed specialized diets for 2 weeks pre- and post-CEA. Groups included control (0 homocystine added, n=9), 1.5 (1.5 g/kg homocystine added, n=10), 3.0 (3.0 g/kg homocystine added, n=9), and 4.5 (4.5 g/kg homocystine added, n=11). The rats underwent a surgical carotid endarterectomy. Endpoints included; plasma homocysteine, intimal hyperplasia, replicative index using with alpha-SM actin and BrdU, hepatic SAM levels, SAH levels, and the hepatic activities of methylenetetrahydrofolate reductase (MTHFR) and cystathionine beta-synthase (CBS). RESULTS: Increasing dietary homocystine produced a proportionate increase in plasma homocysteine and an increase in intimal hyperplasia. Regression analysis of plasma homocysteine levels and intimal hyperplasia showed a significant correlation (r=0.71,P=0.003). Plasma homocysteine levels above 15 microM were associated with significant increases in intimal hyperplasia above 6.5% (P=0.04). Elevation of plasma homocysteine levels to moderate levels (5-25 microM) resulted in significant post-CEA intimal hyperplasia. Cellular analysis of the area of intimal hyperplasia in all diet groups showed comparable amounts of cells positive for alpha-SM actin. However, with increasing levels of dietary homocystine and plasma homocysteine there was an increase in replicative index (P<0.001) as determined by BrdU staining. Increasing dietary homocystine increased plasma homocysteine and was followed by increases in the replicative index thus producing increased intimal hyperplasia and lumenal stenosis. In hepatic measurements the 1.5 and 3.0 g/kg homocystine diets caused: increased liver activity of MTHFR (P=0.03) and decreased hepatic levels of SAM, SAH and SAM/SAH ratios compared to controls. Homocystine treatment did not cause significant alterations in CBS levels (P=0.992). These studies also showed no correlation of the MTHFR and CBS enzymes with plasma homocysteine levels or intimal hyperplasia. However, hepatic levels of SAM showed significant negative correlations with plasma homocysteine (r=-0.58; P=0.006) and with BrdU percentages of cellular proliferation (r=-0.69; P=0.06). CONCLUSION: The degree of post-CEA intimal hyperplasia in a rat model is directly related to the plasma level of homocysteine. The hyperplastic effects of homocysteine may be mediated in part by a physiological insufficiency of methyl donors as shown by decreases in SAM. Thus, increasing levels of plasma homocysteine enhanced and accelerated the smooth muscle cell response after CEA which led to increased intimal hyperplasia and lumenal stenosis. PMID- 11500184 TI - Effect of F-1394, an acyl-CoA:cholesterol acyltransferase inhibitor, on atherosclerosis induced by high cholesterol diet in rabbits. AB - Cholesterol-fed rabbits were used to study the anti-atherosclerotic effect of (1S,2S)-2-[3-(2,2-dimethylpropyl)-3-nonylureido]cyclohexane-1-yl 3-[(4R)-N (2,2,5,5-tetramethyl-1,3-dioxane-4-carbonyl)amino]propionate (F-1394), an acyl CoA:cholesterol acyltransferase (ACAT) inhibitor. To test its effect on the development of atherosclerosis, rabbits were fed a high-cholesterol diet (HCD) for 6 weeks, followed by regular chow (RC) for 12 weeks plus 0 or 100 mg/kg per day F-1394. Serum total cholesterol (TC) rose to approximately 2000 mg/dl on HCD and then declined gradually after the change in diet in both groups. F-1394 significantly reduced the extent of the atherosclerotic lesions and the total and esterified cholesterol contents of the aorta (by 57,38, and 59%, respectively), without affecting the serum TC level. To clarify whether F-1394 accelerates the regression of preexisting atherosclerosis, rabbits were fed HCD for the first 6 weeks and then RC for the next 6 weeks. Then, the rabbits were given 0 or 100 x 2 mg/kg per day F-1394 for another 12 weeks while on RC. F-1394 significantly reduced the extent of the atherosclerotic lesions and the total and esterified cholesterol content in the aorta (by 31, 31, and 43%, respectively), without affecting the serum TC level. These results demonstrate that F-1394 both prevents the formation of atherosclerosis and accelerates its regression without affecting the serum TC level, indicating that F-1394 acts directly on the arterial wall. PMID- 11500185 TI - Distribution of intimal and medial thickening in the human right coronary artery: a study of 17 RCAs. AB - OBJECTIVE: To quantify the distribution of intimal and medial thickening in human right coronary arteries (RCAs) obtained at autopsy. BACKGROUND: The shear and tensile stresses created by arterial bifurcation are believed to result in eccentric fibromuscular intimal thickening that leads to atherosclerosis. Vascular curvature has been cited as a cause of atherosclerosis; however, details of the location and extent of intimal and medial thickness in the largely curved human RCA are not adequately documented. METHODS: The right coronary arteries were obtained from 40 postmortem hearts and cut into 20-30 segments, each being 3 4 mm in length. Microscopic sections from the proximal, acute margin, and distal regions of the RCA were digitized around the circumference of the vessel. Seventeen arteries showed insignificant stenosis (<50%) and were selected for detailed examination. RESULTS: Seventy-one percent (12/17) of proximal sections displayed eccentric intimal thickening. Normalized ensemble averaging revealed a preferential thickening on the myocardial side of the artery. At the acute margin region where curvature is most pronounced and at the distal region, 51% (8/17) of the samples showed eccentric thickening, but the ensemble average thickening in these regions showed no preferential location. In these mildly diseased arteries, the thickened intima comprised of mainly smooth muscle cells with an extracellular matrix of collagen and some elastin. A relatively uniform medial smooth muscle layer was seen at all three locations. CONCLUSIONS: The proximal region of the RCA appears to be a site of intrinsic eccentric intimal thickening with maximum thickness on the myocardial side of the artery. Eccentric thickening does occur in the acute margin and distal regions; however, no distinct pattern or location was evident. PMID- 11500186 TI - Impact of vascular branching sites on focal progression of allograft vasculopathy in transplanted hearts. AB - BACKGROUND: Cardiac allograft vascular disease (CAVD) represents one of the most accelerated progressing coronary syndromes in the human heart. A variety of risk factors have been identified over recent years; however, little is known about the influence of physical forces. As a model for differences in focal blood flow dynamics, we analyzed progression of intimal hyperplasia at vascular bifurcational sites using intravascular ultrasound (IVUS). METHODS: The most diseased vascular sites ("worst sites") in 59 coronary arteries were assessed (30 MHz, motorized pull back) in 25 consecutive heart transplant recipients at baseline (52.8+/-15.3 days postoperatively) and after 1 year of follow up (360.5+/-24.9 days). Progression of intimal hyperplasia was compared between branching and non-branching lesions as well as in focal relation to the position of the flow divider. RESULTS: A total of 41 (69.5%) worst sites were identified at branching locations. Progression of intimal hyperplasia was found to be significantly more severe at bifurcational sites with an increase in plaque area by 1.5+/-1.8 mm(2) in branching versus 0.4+/-0.6 mm(2) in non-branching lesions (P=0.015). The highest rate in focal progression was found at the opposite site of the flow divider with an increase in maximal intimal thickness by 0.3+/-0.23 mm (180 degrees ) as compared to 0.11+/-0.15 mm (90 degrees, P<0.001) and 0.15+/ 0.15 mm (P=0.014) at 270 degrees. CONCLUSIONS: Using serial intravascular ultrasound examinations, vascular branching sites could be identified to be predisposing locations not only for a donor related arteriosclerosis, but also for progression of intimal hyperplasia within transplanted hearts. The highest regional increase in intimal thickness was found at the outer wall of the flow divider, suggesting focal shear or wall stress to be involved in pathogenesis. PMID- 11500187 TI - Vitamin supplementation can markedly reduce the homocysteine elevation induced by fenofibrate. AB - Elevated homocysteine concentrations are a risk factor for atherosclerotic disease. Recently it was reported that lipid lowering with fibrates increases homocysteine by up to 40%. Since elevated homocysteine concentrations can readily be lowered by vitamin supplementation, a randomized, double-blind crossover study was performed to investigate the effect of fenofibrate plus folic acid, vitamin B6 and B12 versus fenofibrate plus placebo in hyperlipidemic men. The crossover study comprised a run-in period of 6 weeks, a first treatment phase of 6 weeks, a washout phase of 8 weeks and a second treatment phase of 6 weeks. Vitamins were given at doses of 650 microg folic acid, 50 microg vitamin B12 and 5 mg vitamin B6 per day for a period of 6 weeks. After fenofibrate plus placebo the increase in homocysteine concentration was 44+/-47%. After fenofibrate plus vitamins it was 13+/-25%, being significantly lower than without vitamins. The increase in homocysteine in response to fenofibrate may counteract the cardioprotective effect of lipid lowering. The addition of vitamins involved in homocysteine metabolism can prevent most of the homocysteine increase seen after fenofibrate plus placebo. Addition of these vitamins to fenofibrate may therefore be warranted for routine use. PMID- 11500188 TI - Impact of postprandial hypertriglyceridemia on vascular responses in patients with coronary artery disease: effects of ACE inhibitors and fibrates. AB - We analyzed vascular responses (endothelial function, oxidant stress) to postprandial hypertriglyceridemia (PHTG) in patients with coronary artery disease (CAD) to reveal potential therapeutical effects of angiotensin converting enzyme inhibition (ACE-I) and of lipid lowering (fibrate). The study population (n=39, mean age: 60 years) consisted of four groups, all of which had angiographically documented CAD. A high fat group (n=9) consumed a high fat meal, a low fat group (n=9) a low fat meal, and ACE-I (n=10) or fibrate (n=11) groups consumed a high fat meal plus lisinopril or fenofibrate. Serum triglycerides (TG) increased significantly 2 h after eating a test meal in all groups with the exception of the low fat group. In the high and low fat groups changes of serum TG were positively correlated (r=0.664, P<0.005) with changes of phorbol ester-activated leukocyte superoxide anion radical (O(2-.)) formation and were negatively correlated (r=-0.488, P<0.05) with flow-mediated brachial artery dilation (FMD). There was a negative correlation (r=-0.419, P=0.094) between FMD and changes of O(2-.) formation in the high and low fat groups. In the ACE-I and fibrate groups, O(2-.) formation decreased 2 h after eating a test meal (from 5.34+/-1.01 to 3.81+/-1.15 nmol/10(6)cells per min, P<0.01, and from 4.66+/-0.91 to 4.26+/-0.97 nmol/10(6)cells per min, P=0.374, respectively). However, endothelial function did not show any significant changes 2 h after eating a test meal in all groups. PHTG increases oxidant stress and further deteriorates endothelial function, even in patients with CAD. Both ACE-I and fibrates have an antioxidant effect but no acute beneficial effects in terms of endothelial function under conditions of PHTG in CAD patients. PMID- 11500189 TI - Association of the Sst-I polymorphism at the APOC3 gene locus with variations in lipid levels, lipoprotein subclass profiles and coronary heart disease risk: the Framingham offspring study. AB - Apolipoprotein (apo) CIII participates in the regulation of the metabolism of triglyceride-rich lipoproteins and it is a major component of chylomicrons and VLDL. The APOC3 gene is on chromosome 11q23 and is highly polymorphic. The less common allele (S2) of the SstI polymorphism on the 3' untranslated region of the APOC3 gene has been previously associated with increased triglycerides, total cholesterol (TC), and apoCIII levels and cardiovascular risk on several, but not all, studies. The aim of this study was to examine the association of this polymorphism with plasma lipid levels, lipoprotein subfractions and coronary heart disease (CHD) risk in a population-based study: The Framingham Offspring Study. The frequency of the S2 allele was 0.086, consistent with previous reports in Caucasian populations. In men, the S2 allele was associated with lower concentrations of high-density lipoprotein cholesterol (HDL-C; P<0.04) and HDL2-C (P<0.02) and a significant increase in apoCIII non-HDL (P<0.05). TG levels were higher in men carriers of the S2 allele, but this association did not reach statistical significance (P=0.30). Conversely, in women, the S2 allele was associated with increased TC (P<0.03), low-density lipoprotein cholesterol (LDL C; P<0.03), and ApoB levels (P<0.04). Lipoproteins subfractions were also examined using nuclear magnetic resonance (NMR) spectroscopy. S2 male carriers had significantly lower concentrations of large LDL and a significant reduction in LDL particle size (P<0.04). In women, there was a significant increase in intermediate LDL particles (P<0.05) with no significant effect on lipoprotein diameters. We also examined the associations between the S2 allele and biochemical markers of glucose metabolism. In men, the S2 allele was associated with elevated fasting insulin concentrations (P<0.04), whereas no significant associations were observed in women. Despite the described associations with lipid and glucose metabolism related risk factors, we did not find any significant increase in CHD risk associated with the S2 allele in this population. PMID- 11500190 TI - Apolipoprotein E genotype affects plasma lipid response to atorvastatin in a gender specific manner. AB - The response to therapy with hypolipidemic agents shows considerable individual variation. These differences may be due to the interaction of environmental and genetic factors that affect drug bioavailability, receptor function or ligand structure. Our objective was to assess the effect of apolipoprotein (apo) E genotype and gender on lipid-lowering response to the HMG CoA reductase inhibitor, atorvastatin. Genotyping was carried out on DNA from 328 male and female subjects who participated in a multicentric, double-blind clinical trial, and received 10 mg/day of atorvastatin. Our data demonstrate no significant gender differences for LDL cholesterol levels at baseline. Moreover, mean LDL-C lowering was similar in men (-36.2%, range -2.7 to -57.8%) and in women (-38.1%, range -9.5 to -58.5%) as compared to baseline. However, men carrying the epsilon2 allele had a significantly higher mean LDL-C response (-44%) than epsilon3 homozygotes (-37%) and epsilon4 carriers (-34%); P=0.01 for apoE group by treatment interaction. No such gene/treatment interactions were noted in women, with those carrying the epsilon2 allele showing a similar mean response (-34%) as epsilon3 homozygotes (-39%) and epsilon4 carriers (-34%). Mean plasma triglyceride lowering with atorvastatin was 17%. A significant apoE group by treatment interaction (P=0.010) was also observed in men, with epsilon2 carriers being more responsive (-27%) than epsilon3/3 (-13%) and epsilon4 (-22%). This interaction was not observed in women. In summary, atorvastatin treatment had similar effects on plasma lipid levels in both men and women; however, the apoE gene locus was a significant predictor of LDL-C and TG responses to atorvastatin therapy in men, but not in women. PMID- 11500191 TI - Pomegranate juice consumption inhibits serum angiotensin converting enzyme activity and reduces systolic blood pressure. AB - Consumption of pomegranate juice which is rich in tannins, possess anti atherosclerotic properties which could be related to its potent anti-oxidative characteristics. As some antioxidants were recently shown to reduce blood pressure, we studied the effect of pomegranate juice consumption (50 ml, 1.5mmol of total polyphenols per day, for 2 weeks) by hypertensive patients on their blood pressure and on serum angiotensin converting enzyme (ACE) activity. A 36% decrement in serum ACE activity and a 5% reduction in systolic blood pressure were noted. Similar dose-dependent inhibitory effect (31%) of pomegranate juice on serum ACE activity was observed also in vitro. As reduction in serum ACE activity, even with no decrement in blood pressure, was previously shown to attenuate atherosclerosis, pomegranate juice can offer a wide protection against cardiovascular diseases which could be related to its inhibitory effect on oxidative stress and on serum ACE activity. PMID- 11500192 TI - Genetic variation at the lipoprotein lipase locus and plasma lipoprotein and insulin levels in the Quebec Family Study. AB - The associations between the S447X, BamHI, HindIII and PvuII DNA variants of the lipoprotein lipase (LPL) gene and indicators of body fat, fat distribution and plasma lipids and insulin were studied in the Quebec Family Study cohort. Strong linkage disequilibrium among all the markers was observed. For the S447X polymorphism, plasma very low density lipoprotein (VLDL)-cholesterol (chol) (P<0.001), total triglyceride (TG) (P=0.033) and VLDL-TG (P<0.001) levels were lower and high density lipoprotein (HDL)-chol level higher (P<0.001) in the subjects homozygous or heterozygous for X447 (X447+, n=160) compared to the homozygotes for the S447 allele (X447-, n=576). The BamHI, PvuII and HindIII polymorphisms were not associated with the plasma lipid values when all X447 allele carriers were removed. In addition, the HindIII polymorphism as well as the HindIII and S447X markers combination influenced the insulin area under the curve during an oral glucose tolerance test. We conclude that DNA sequence variation in the LPL gene contributes significantly to the variability in the levels of VLDL-chol, total- and VLDL-TG as well as HDL-chol. The effects of the other polymorphisms considered here are most likely mediated by their linkage disequilibrium with the S447X mutation. In addition, genetic variation at the LPL locus may, by an unknown mechanism, influence insulin metabolism but not body fat variability. PMID- 11500193 TI - Femoral artery wall thickness and stiffness in evaluation of peripheral vascular disease in type 2 diabetes mellitus. AB - Stiffening and thickening of arterial wall are two important components of atherosclerosis. The purpose of this study was to evaluate the effects of femoral artery wall stiffness on clinical manifestation of peripheral vascular disease (PVD) in type 2 diabetes mellitus. The subjects were 315 patients with type 2 diabetes. Presence of intermittent claudication and/or leg pain at rest and reduced ankle-brachial blood pressure index (ABI<0.9) were used as a subjective and an objective index of PVD, respectively. Femoral artery intima-media thickness (FA-IMT) and stiffness parameter beta (FA-stiffness beta) were measured by ultrasound methods. Symptomatic patients (N=58) showed greater values for both FA-IMT and FA-stiffness beta than those without symptom (N=257). Similarly, patients with reduced ABI (N=56) had greater FA-IMT and FA-stiffness beta than those without (N=259). However, correlation between FA-IMT and FA-stiffness beta was not impressive, especially in the symptomatic patients. To evaluate the effect of FA-stiffness beta on PVD symptoms, the subjects were divided into three subgroups according to FA-IMT, and then FA-stiffness beta was compared between those with and without PVD symptoms in each subgroup. The symptomatic patients had greater FA-stiffness beta values than the asymptomatic subjects in all the three subgroups. Multiple logistic regression analysis indicated that the presence of PVD symptoms was associated more closely with increased FA-stiffness beta than with increased FA-IMT, whereas reduced ABI was associated more closely with FA-IMT than with FA-stiffness beta. These data suggest that stiffening of arterial wall has a significant impact on PVD manifestations, particularly on the leg symptoms, in patients with type 2 diabetes. PMID- 11500194 TI - Immunolocalization of apolipoproteins in aortic atherosclerosis in American youths and young adults: findings from the PDAY study. AB - The immunohistochemical distribution of apolipoproteins in the abdominal aortas of 142 men, 15-34 years of age, collected in a cooperative multicenter study group (Pathobiological Determinants of Atherosclerosis in Youth) was examined in relationship to serum VLDL+LDL+HDL cholesterol levels. ApoB deposits were limited to the intima of specimens with intimal fibro cellular thickening or atherosclerotic lesions. Apo A-I, E and J were observed in both the intima and media of the aortas with intimal lesions. The pattern of apoJ distribution was similar to that of apoA-I and E. The distribution patterns of these apolipoproteins in these young adults were very similar to those in adults and old men seen in an earlier study. The extent of apolipoprotein distribution in the intima and media increased with age and the stage of atherosclerosis development, but was not correlated significantly with serum VLDL+LDL or HDL cholesterol levels. The infiltration of lipoprotein particles into the aortic wall seems to be more strongly associated with the progression of intimal lesions rather than with serum cholesterol levels. PMID- 11500195 TI - Effect of glutathione S-transferase M1 genotype on progression of atherosclerosis in lifelong male smokers. AB - We hypothesize that smokers with the null genotype for GSTM1 (GSTM1-0), who thus lack the detoxification enzyme glutathione S-transferase mu-1, develop atherosclerosis at an increased rate compared to smokers with the positive genotype (GSTM1-1). We used data from a 2-year randomized placebo-controlled trial on the effect of vitamin E on atherosclerosis among 189 male smokers. Progression of atherosclerosis was measured by 2-year change of the common carotid intima media thickness (CCA-IMT) as measured by B-mode ultrasonography. The frequency of GSTM1-0 genotype was 0.5 in both the placebo and the vitamin E group. Smokers with GSTM1-0 genotype had a tendency to higher baseline CCA-IMT values than those with GSTM1-1 (0.97 versus 0.92 mm, P=0.09). Within the placebo group, more CCA-IMT progression was found for smokers with the GSTM1-0 than for smokers with the GSTM1-1 genotype after adjustment for baseline IMT and major CVD risk factors (0.050 versus -0.002 mm, P=0.046). In the vitamin E group no effect of GSTM1 genotype on atherosclerosis progression was found. Overall, smokers with GSTM1-0 genotype had a higher mean 2-year progression compared to those with GSTM1-1 as shown by a difference in increase of 0.042 mm (95% CI 0.006; 0.078, P=0.02). In conclusion, our data suggest that smokers lacking the detoxifying enzyme GST mu-1 develop progression of atherosclerosis at an increased rate. PMID- 11500196 TI - Involvement of polymorphisms in the chemokine system in the susceptibility for coronary artery disease (CAD). Coincidence of elevated Lp(a) and MCP-1 -2518 G/G genotype in CAD patients. AB - The central role of chemokines in the pathogenesis of atherosclerosis has been made clear. Recently polymorphisms in the gene regulatory region of MCP-1 and in the promoter region of RANTES have been found, which increase the expression of these chemokines. We investigated the role of these polymorphisms together with the chemokine SDF-1-801A and the chemokine receptors CCR2-64I and CCR5Delta32 mutations in 318 patients with coronary artery disease (CAD) referred to coronary bypass surgery, comparing them with 320 healthy controls. The prevalence of the MCP-1 -2518 G/G homozygotes was significantly higher among CAD patients than among controls (P<0.005; OR=2.2 (95% CI 1.25-3.92). The Lp(a) levels of CAD patients with G/G genotype were significantly higher than those in patients with G/A or A/A genotypes. No CAD patients homozygous for the CCR5Delta32 and CCR2-64I mutations have been found. The genotype distributions of the two alleles deviated from the Hardy Weinberg equilibrium in patients, indicating that the numbers of homozygotes were significantly lower than expected. The MCP-1 -2518G variant in homozygous form appears as a genetic risk factor for severe CAD. This genotype is associated with elevated Lp(a) levels in patients. Individuals homozygous for CCR2-64I or CCR5Delta32 mutations are at reduced risk for severe CAD. PMID- 11500197 TI - Autoantibodies and endothelial dysfunction in well-controlled, uncomplicated insulin-dependent diabetes mellitus patients. AB - Patients with insulin-dependent diabetes mellitus (IDDM) are well known to be at high risk of vascular disease, and dysfunction of vascular endothelium is considered as an early step in the development of diabetic complications. Because of the involvement of autoimmunity in the pathogenesis of IDDM, our aim was to assess, in 45 IDDM patients without clinically evident vascular complications, whether early signs of endothelial cell dysfunction were correlated to alterations of the immune system. IDDM patients were characterized by significantly increased serum levels of C-reactive protein, of polymorphonuclear cells-derived elastase, of endothelin-1 (ET-1) and of thrombomodulin, while plasma concentrations of fibronectin (FNT) were significantly decreased, with a statistically significant inverse correlation between ET-1 and FNT values. The presence of circulating immune complexes (CIC) was investigated in 36 out of our 45 IDDM patients, and values above the cut-off were found in 17 (47.2%) of them. One-third of all patients showed values above the cut-off for IgG-aCL. In IDDM patients, at variance from the control group, the levels of ET-1 were directly correlated to those of von Willebrand factor, of anticardiolipin beta(2)-GPI and of CIC, with an inverse correlation with plasma FNT. An association between antiphospholipid antibodies and endothelial dysfunction and/or activation is therefore suggested, pointing to a synergism, in the early phases of IDDM vascular disease, between generation of autoantibodies and endothelial activation. PMID- 11500198 TI - Vascular dilatory functions of ovo-lactovegetarians compared with omnivores. AB - Vegetarians have lower blood pressure and lower cardiovascular mortality. Vegetarian diets may have lower cardiovascular risks through positive influence on endothelium-dependent relaxation and related functions. The objectives of this study were to assess the differences of vascular dilatory functions between middle-aged vegetarians and sex and age-matched omnivores before they develop any clinical manifestations of atherosclerosis. Twenty healthy vegetarians over the age of 50 and 20 healthy omnivores over the age of 50 were recruited for this study. Subjects with known risk factors for atherosclerosis such as hypertension, diabetes, obesity, hypercholesteremia, cigarette smoking, family history of vascular diseases, or taking any regular medication were excluded. Medical history, body weight, height, and duration of vegetarian diet were recorded. Baseline CBC, urinalysis and biochemical data such as fasting blood glucose, thyroid function, blood urea nitrogen, creatinine, serum electrolytes (sodium, potassium, chloride, calcium and magnesium), lipid profiles [total cholesterol, triglycerides, high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol] were obtained after a 14 h fast. Blood pressures and heart rate were recorded in supine position. Vascular dilatory functions, both flow-mediated (endothelium-dependent) and nitroglycerin-induced (endothelium independent), were evaluated by using a non-invasive ultrasonographic method. The results show that there were no significant differences in the baseline characteristic between the vegetarians and the omnivores. There were also no significant differences in serum glucose, lipid profiles and thyroid function between these two groups. However, vasodilatation responses (both flow-mediated and nitroglycerin-induced) were significantly better in the vegetarian group and the degree of vasodilatation appeared to be correlated with years on vegetarian diets. Our findings suggest that vegetarian diets, by themselves, have a direct beneficial effect on vascular endothelial and smooth muscle function and may help to account for the lower incidence of atherosclerosis and cardiovascular mortality. PMID- 11500199 TI - Familial defective apolipoprotein B-100: a mutation emerged in the Mesolithic period. PMID- 11500200 TI - Biotechnology in the textile industry--perspectives for the new millennium. PMID- 11500201 TI - Biodegradation of bioaccessible textile azo dyes by Phanerochaete chrysosporium. AB - Azo dyes are important chemical pollutants of industrial origin. Textile azo dyes with bioaccessible groups for lignin degrading fungi, such as 2-methoxyphenol (guaiacol) and 2,6-dimethoxyphenol (syringol), were synthesised using different aminobenzoic and aminosulphonic acids as diazo components. The inocula of the best biodegradation assays were obtained from a pre-growth medium (PAM), containing one of the synthesised dyes. The results of the dye biodegradation assays were evaluated every 7 days, by the decrease of the absorbance at the maximum wavelength of the dye, by the decrease of the sucrose concentration in the culture medium and by the increase of the biomass during the 28 days of assay. It was observed that the extent of dye biodegradation depended on the sucrose concentration, on the degraded dye structure and, on the dye present in the PAM medium. PMID- 11500202 TI - A packed-bed fungal bioreactor for the continuous decolourisation of azo-dyes (Orange II). AB - The degradation of an azo dye, Orange II, by immobilised Phanerochaete chrysosporium in a continuous packed bed bioreactor for periods longer than 30 days has been carried out. Nearly complete decolourisation (>95%) was achieved when working at a high dye load rate of 0.2 g x l(-1) x d(-1), a temperature of 37 degrees C, a hydraulic retention time (HRT) of 24 h and applying oxygen gas in a pulsed flow. These conditions allowed Manganese peroxidase (MnP) production and the subsequently Orange II decolourisation. A correlation between residual MnP activity in the effluent and decolourisation was established. Apparently, for decolourisation to be effective, a minimum MnP activity was required, no substantial increase in efficiency at MnP activities higher than 10 U x 1(-1) was observed. The treatment caused, the breakdown of the chromophoric group as well as the cleavage of the aromatic ring. PMID- 11500203 TI - Decolourisation of Remazol Brilliant Blue R via a novel Bjerkandera sp. strain. AB - A novel strain of Bjerkandera sp. (B33/3), with particularly high decolourisation activities upon Poly R-478 and Remazol Brilliant Blue R (RBBR) dyes, was isolated. The role of the ligninolytic extracellular enzymes produced by this strain on decolourisation of RBBR was studied in some depth. The basis of decolourisation is an enzyme-mediated process, in which the main enzyme responsible is a recently described peroxidase with capacity for oxidation of manganese, as well as veratryl alcohol and 2,6-dimethoxyphenol in a manganese independent reaction. PMID- 11500204 TI - Capacity of Irpex lacteus and Pleurotus ostreatus for decolorization of chemically different dyes. AB - The rate and efficiency of decolorization of poly R-478- or Remazol Brilliant Blue R (RBBR)-containing agar plates (200 microg x g(-1)) were tested to evaluate the dye degradation activity in a total of 103 wood-rotting fungal strains. Best strains were able to completely decolorize plates within 10 days at 28 degrees C. Irpex lacteus and Pleurotus ostreatus were selected and used for degradation of six different groups of dyes (azo, diazo, anthraquinone-based, heterocyclic, triphenylmethane, phthalocyanine) on agar plates. Both fungi efficiently degraded dyes from all groups. Removal of RBBR, Bromophenol blue, Cu-phthalocyanine, Methyl red and Congo red was studied with I. lacteus also in liquid medium. Within 14 days, the following color reductions were attained: RBBR 93%, Bromophenol blue 100%, Cu-phthalocyanine 98%, Methyl red 56%, Congo red 58%. The ability of I. lacteus to degrade RBBR spiked into sterile soil was checked, the removal being 77% of the dye added within 6 weeks. The capacity of selected white rot fungal species to remove efficiently diverse synthetic dyes from water and soil environments is documented. PMID- 11500205 TI - Use of laccase together with redox mediators to decolourize Remazol Brilliant Blue R. AB - A pure fungal laccase, obtained from a commercial formulation used in the textile industry, did not decolourize Remazol Brilliant Blue R (RBBR). Decolourization was only observed when a small molecular weight redox mediator was added together with the laccase. Under the conditions specified, violuric acid (5.7 mM) was the most effective mediator studied and almost complete decolourization was observed within 20 min. In contrast, 1-hydroxybenzotriazole (HOBT, 11 mM) decolourized RBBR at about a two-fold slower rate and to a lesser extent. Also, higher concentrations of HOBT were inhibitory which could be due to inactivation of laccase by the toxic HOBT radical. The commercial laccase formulation that contained phenothiazine-10-propionic acid as the mediator was least effective, giving 30% decolourization under equivalent conditions. We suggest that similar laccase plus mediator systems could be used for the detoxification of related anthraquinone textile dyes. PMID- 11500206 TI - Indigo degradation with purified laccases from Trametes hirsuta and Sclerotium rolfsii. AB - The degradation of the textile dye indigo with purified laccases from the fungi Trametes hirsuta (THL1 and THL2) and Sclerotium rolfsii (SRL1) was studied. All laccases were able to oxidize indigo yielding isatin (indole-2,3-dione), which was further decomposed to anthranilic acid (2-aminobenzoic acid). Based on the oxygen consumption rate of the laccases during indigo degradation, a potential mechanism for the oxidation of indigo involving the step-wise abstraction of four electrons from indigo by the enzyme was suggested. Comparing the effect of the known redox-mediators acetosyringone, 1-hydroxybenzotriazole (HOBT) and 4 hydroxybenzenesulfonic acid (PHBS) on laccase-catalyzed degradation of indigo, we found a maximum of about 30% increase in the oxidation rate of indigo with SRL1 and acetosyringone. The particle size of indigo agglomerates after laccase treatment was influenced by the origin of the laccase preparation and by the incubation time. Diameter distributions were found to have one maximum and compared to the indigo particle size distribution of the control, for all laccases, the indigo agglomerates seemed to have shifted to smaller diameters. Bleaching of fabrics by the laccases (based on K/S values) correlated with the release of indigo degradation products. PMID- 11500207 TI - Decolorization of textile indigo dye by ligninolytic fungi. AB - The indigo dye is extensively used by textile industries and is considered a recalcitrant substance, which causes environmental concern. Chemical products used on textile processing, which affect the environment through effluents, can be voluminous, colored and varied. Vat textile dyes, like indigo, are often used and dye mainly cellulosic fibers of cotton. Decolorization of this dye in liquid medium was tested with ligninolytic basidiomycete fungi from Brazil. Decolorization started in a few hours and after 4 days the removal of dye by Phellinus gilvus culture was in 100%, by Pleurotus sajor-caju 94%, by Pycnoporus sanguineus 91% and by Phanerochaete chrysosporium 75%. No color decrease was observed in a sterile control. Thin layer chromatography of fungi culture extracts revealed only one unknown metabolite of Rf=0.60, as a result of dye degradation. PMID- 11500208 TI - Thermo-alkali-stable catalases from newly isolated Bacillus sp. for the treatment and recycling of textile bleaching effluents. AB - Three thermoalkaliphilic bacteria, which were grown at pH 9.3-10 and 60-65 degrees C were isolated out of a textile wastewater drain. The unknown micro organisms were identified as thermoalkaliphilic Bacillus sp. Growth conditions were studied and catalase activities and stabilities compared. Catalases from Bacillus SF showed high stabilities at 60 degrees C and pH 9 (t1/2=38 h) and thus this strain was chosen for further investigations, such as electron microscopy, immobilization of catalase and hydrogen peroxide degradation studies. Degradation of hydrogen peroxide with an immobilized catalase from Bacillus SF enabled the reuse of the water for the dyeing process. In contrast, application of the free enzyme for treatment of bleaching effluents, caused interaction between the denaturated protein and the dye, resulting in reduced dye uptake, and a higher color difference of 1.3DeltaE* of dyed fabrics compared to 0.9DeltaE* when using the immobilized enzyme. PMID- 11500209 TI - Batch tests for assessing decolourisation of azo dyes by methanogenic and mixed cultures. AB - Most of the published studies on azo dye colour removal involve anaerobic mixed cultures and there is some interest in the knowledge of how dye reduction occurs, if by facultative, strictly anaerobic or both bacterial trophic groups present in classic anaerobic digestors. This paper describes the behaviour of methanogenic and mixed bacteria cultures on the colour removal in batch systems, of a commercial azo dye, C.I. Acid Orange 7, used in paper and textile industries. The aim of this study is to demonstrate, by analysing dye decolourisation, that it occurs with mixed cultures as well as with strictly anaerobic (methanogenic) cultures. Tests were performed with a range of dye concentrations between 60 and 300 mg x l(-1). The influence of dye concentration on the carbon source removal and decolourisation processes was studied. The effect of carbon source concentration on colour removal was also analysed for both cultures. The degradation rates in mixed and methanogenic cultures were compared. The consumption of carbon source was monitored by COD analysis and dye degradation by ultraviolet-visible spectrophotometry and thin layer chromatography. PMID- 11500210 TI - Effect of some operational parameters on textile dye biodegradation in a sequential batch reactor. AB - The combination of anaerobic and aerobic periods in the operation cycle of a Sequencing Batch Reactor (SBR) was chosen to study biological color removal from simulated textile effluents containing reactive, sulfonated, monoazo and diazo dyes, respectively, Remazol Brilliant Violet 5R and Remazol Black B. 90% color removal was obtained for the violet dye in a 24-h cycle with a Sludge Retention Time (SRT) of 15 days and an aerated reaction phase of 10 h. For the black dye only 75% color removal was achieved with the same operational conditions and no improvement was observed with the increase of the SRT to 20 days. For the violet dye a reduction of the color removal values from 90 to 75% was observed with the increase of the aerated reaction phase from 10 to 12 h. However, this increase did not promote the aerobic biodegradation of the produced aromatic amines. Abiotic tests were performed with sterilized SBR samples and no color removal was observed in cell-free supernatants. However color removal values of 30 and 12% were observed in the presence of sterilized cells and supernatants with violet and black dye, respectively and could be attributed to the presence of active reducing principles in the sterilized samples. PMID- 11500211 TI - Biodegradation, decolourisation and detoxification of textile wastewater enhanced by advanced oxidation processes. AB - Recently, an increasing application of so called advanced oxidation processes (AOPs) to industrial wastewater has been observed. In particular, an integrated approach of biological and chemical treatment of wastewater is advantageous conceptually. The subject of our study was synthetic wastewater, simulating effluents from knitting industry. The wastewater contained components that are very often used in Polish textile industry: an anionic detergent Awiwaz KG conc., a softening agent Tetrapol CLB and an anthraquinone dyestuff-Acid Blue 40, CI 2125. The toxicity of the detergents and the dye was determined in terms of effective concentration EC50 using mixed cultures of activated sludge as well as pure culture of luminescent bacteria Vibrio fischerii NRRLB-11177. The dye did not undergo biodegradation without AOPs pretreatment, therefore a degree of its removal (decolourisation) by the AOPs has been determined and its bio-sorption properties on the flocks of activated sludge have been studied. The dye adsorption onto flocks of activated sludge was described by Henry's isotherm. Our investigations focussed on the influence of various oxidants like O3, H2O2 and UV light on biodegradation of single components aqueous solution as well as of the whole textile wastewater. The results of kinetic measurements of the biodegradation (by means of acclimated activated sludge) was described by Monod type of kinetic equation. The experimental evidence of the positive effect of chemical oxidation pretreatment on the biodegradation of recalcitrant compounds was quantified by estimation of the kinetic parameters of the Monod equation. Due to the AOPs pretreatment a decrease of the Monod constant and an increase of maximal specific growth rate was observed. The activity of degradative enzymes of activated sludge was assayed by the methods of 2-[4-iodophenyl]-3-[4-nitrophenyl] 5-phenyltetrazolium chloride test. PMID- 11500212 TI - Integration of biotechnological wastewater treatment units in textile finishing factories: from end of the pipe solutions to combined production and wastewater treatment units. AB - Increasing costs for water, wastewater and energy put pressure on textile finishing plants to increase the efficiency of wet processing. An improved water management can decrease the use of these resources and is a prerequisite for the integration of an efficient, anaerobic on-site pretreatment of effluents that will further cut wastewater costs. A two-phase anaerobic treatment is proposed, and successful laboratory experiments with model effluents from the cotton finishing industry are reported. The chemical oxygen demand of this wastewater was reduced by over 88% at retention times of 1 day or longer. The next step to boost the efficiency is to combine the production and wastewater treatment. The example of cotton fabric desizing (removing size from the fabric) illustrates how this final step of integration uses the acidic phase bioreactor as a part of the production and allows to close the water cycle of the system. PMID- 11500213 TI - Enzyme-retting of flax and characterization of processed fibers. AB - Enzyme-retting formulations consisting of Viscozyme L, a pectinase-rich commercial enzyme product, and ethylenediaminetetraacetic acid (EDTA) were tested on Ariane fiber flax and North Dakota seed flax straw residue. Flax stems that were crimped to disrupt the outer layers were soaked with various proportions of Viscozyme-EDTA solutions, retted, and then cleaned and cottonized with commercial processing equipment. Fiber properties were determined and crude test yarns were made of raw and Shirley cleaned flax fibers and cotton in various blend levels. Cleaned fibers were obtained from both seed and fiber flax types, but with variations due to treatment. Retting formulations produced fibers having different properties, with enzyme levels of 0.3% (v/v as supplied) giving finer but weaker fibers than 0.05% regardless of EDTA level. Experimental yarns of blended flax and cotton fibers varied in mass coefficient of variation, single end strength, and nep imperfections due to sample and formulation. With cost and fiber and yarn quality as criteria, results established a range in the amounts of components comprising retting formulations as a basis for further studies to optimize enzyme-retting formulations for flax. Under conditions examined herein, Viscozyme L at 0.3% (v/v) plus 25 mM EDTA produced the best test yarns and, therefore, established a base for future studies to develop commercial-grade, short staple flax fibers for use in textiles. PMID- 11500214 TI - Analysis of retted and non retted flax fibres by chemical and enzymatic means. AB - Flax fibres (Linum usitatissimum L.) were subjected to chemical and enzymatic analysis in order to determine the compositional changes brought about by the retting process and also to determine the accessibility of the fibre polymers to enzymatic treatment. Chemical analysis involved subjecting both retted and non retted fibres to a series of sequential chemical extractions with 1% ammonium oxalate, 0.05 M KOH, 1 M KOH and 4 M KOH. Retting was shown to cause minimal weight loss from the fibres but caused significant changes to the pectic polymers present. Retted fibres were shown to have significantly lower amounts of rhamnogalacturonan as well as arabinan and xylan. In addition the average molecular mass of the pectic extracts was considerably lowered. Enzyme treatment of the 1 M KOH extracts with two different enzymes demonstrated that the non retted extract contained a relatively high molecular weight xylan not found in the retted extract. Treatment of the 1 M KOH extracts and the fibres with Endoglucanase V from Trichoderma viride demonstrated that while this enzyme solubilised cellulose as well as xylan and xyloglucan oligomers from the extract, it had limited access to these polymers on the fibre. MALDI-TOF MS analysis of the material solubilised from the extract suggested that the xylan was randomly substituted with 4-O-methyl glucuronic acid moieties. The xyloglucan was shown to be of the XXXG type and was substituted with galactose and fucose units. The enzyme treatments of the fibres demonstrated that the xylan and xyloglucan polymers in the fibres were not accessible to the enzyme but that material which was entrapped by the cellulose could be released by the hydrolysis of this cellulose. PMID- 11500215 TI - Mechanism of interaction between cellulase action and applied shear force, an hypothesis. AB - An overview is given of what is known in literature concerning the structure of both cellulose and cellulase enzymes and the enzymatic degradation of cellulose. Based on this knowledge, a hypothesis is formulated about the relation between cellulase performance and required applied shear force on the fabric. In short, the specific cellulase activity is highest on the flexible amorphous cellulose when compared to that with the more rigid crystalline cellulose. When applying cellulase activity on damaged, fuzzy fabric, the connection point of cellulose material oriented away from the fiber axis, which is partly amorphous due to the damage, will be turned again to a more rigid, mainly crystalline structure. Due to this shift, this connection point will function as a primary point of application for shear force, resulting in removal of the fuzz. PMID- 11500216 TI - Guidelines to come to minimized tensile strength loss upon cellulase application. AB - Application of cellulase technology in the textile production process often results in a certain loss of tensile strength along with the desired performance. In this paper guidelines are given how to come to minimization or even prevention of tensile strength loss. Part of the considerations is based on the hypothesis given in the accompanying paper (Lenting and Warmoeskerken, 2001, J. Biotechnol.) concerning the mechanism of interaction between cellulase action and applied shear force. Recommendations given concern the enzyme choice, process parameters and enzyme targeting. PMID- 11500217 TI - Application of microassays for investigation of cellulase abrasive activity and backstaining. AB - Model microassays were used for testing the denim-washing performance and indigo backstaining for Trichoderma reesei and Chrysosporium lucknowense commercial cellulase preparations on a 'test-tube scale'. C. lucknowense preparation demonstrated a higher potential in the denim biostoning process. The performance of four purified cellulases (two endoglucanases and two cellobiohydrolases) from C. lucknowense on cotton textiles was assayed, and the key enzyme (endoglucanase with a molecular weight of 25 kDa) responsible for high abrasion effects on denim fabrics was found. PMID- 11500218 TI - Cellulase finishing of woven, cotton fabrics in jet and winch machines. AB - Some authors have reported that as the applied agitation rate increases, the apparent activity of the endoglucanases from Trichoderma reesei towards cotton cellulose increases more markedly than does the apparent activity of the cellobiohydrolases. This suggests that the quality of cellulase finishing effects on cellulosic textiles may be machine-type dependent. The present work using total crude, endoglucanase-rich and cellobiohydrolase-rich cellulases from T. reesei confirmed that the final properties of woven, cotton fabrics treated under realistic processing conditions in a jet machine, were measurably and perceivably different from those of the same fabrics, treated using the same processing conditions of temperature, time, pH, enzyme concentration and fabric to liquor ratio, but in a winch machine. The results are interpreted in terms of the effects of agitation rate on the adsorption-desorption behaviour of the T. reesei endoglucanases and cellobiohydrolases. PMID- 11500219 TI - Action of purified Trichoderma reesei cellulases on cotton fibers and yarn. AB - In this work the possibility and potential of treating cotton fibers and yarns instead of fabrics with monocomponent cellulases was investigated. Different pretreatments on fibers were performed and tested in order to improve the accessibility of cotton to enzymatic modification. The enzymatic treatments were evaluated microscopically and by analysing the effects of treated fibers on spinnability, yarn evenness, tenacity and pilling. The accessibility of the cotton fibers for cellulases could be increased by different pretreatments. Steaming of fibers prior to enzymatic treatment was found to be an efficient way to increase hydrolysis levels. Cellulase treatments of carded yarns resulted in modification of yarn properties. Decrease in yarn hairiness was observed and the knitted fabric made of the treated yarn showed a lowered tendency towards pilling. In all cases endoglucanase activity rather than cellobiohydrolase activity was responsible for these modifications. PMID- 11500220 TI - Biostoning of denims by Penicillium occitanis (Pol6) cellulases. AB - The Pol6 mutant of Penicillium occitanis, secreting a large quantity of cellulases, was cultivated in fermentor using a local paper pulp as an inducer substrate. A high titer of extracellular cellulase activity was reached after a fed batch process: 23 IU x ml(-1) filter paper activity, 21 IU x ml(-1) CMCases activity (endoglucanase units) and 25 mg x ml(-1) of proteins. Various tests were done to compare the action of the P. occitanis cellulases with those commercially available and with the traditional stonewashing process. This cellulase preparation was successfully applied in a biostoning process at an industrial scale. The abrasive effect of the P. occitanis cellulases was very uniform and with an efficiency comparable to that obtained by the commercial ones. PMID- 11500221 TI - Enzyme processing of textiles in reverse micellar solution. AB - Scouring of cotton using pectinase enzyme, bioscouring, in reverse micellar system was studied. The effectiveness of bioscouring was evaluated by measuring weight loss of cotton, analyzing pectin and cotton wax remaining and by wetness testing. Pectinase enzyme showed excellent activity even in organic media, and the effectiveness of scouring was equivalent or better than that achieved by conventional alkaline process or bioscouring in aqueous media. Enzymatic modification of wool using protease enzyme in the same system was also studied. It has found that felting property and tensile strength of wool fabrics treated by protease in reverse micellar system were superior to those in aqueous media. Possibilities of utilization of the same system for the subsequent textile dyeing process were also investigated. It was found that cotton and polyester fabrics were dyed satisfactorily by reverse micellar system compared to conventional aqueous system. PMID- 11500223 TI - Extremozymes for improving wool properties. AB - The project 'EXTRETEX' funded by the German Federal Foundation Environment (DBU, Osnabruck, Germany) aims at the improvement of wool properties dyeability, handle, felting behaviour and degree of whiteness by means of enzymes derived from extremophilic micro-organisms. In this paper the effects of a commercial thermo- and alkalistable protease on wool with regard to the degree of whiteness, the dyeability and the felting behaviour are presented. A method to treat wool top and wool fabric was developed on a laboratory scale in which the protease was integrated into the pre-washing step of a dyeing process. This treatment method was than scaled up and tested on an industrial winch beck for fabric. With this method-the addition of enzyme in the pre-washing step-the degree of whiteness is generally enhanced. Dyeing untreated and the enzyme-treated wool with Lanasol Blue 8G leads to an improved dyestuff uptake and a distinctive difference in the colour shade for the latter. Microscopy pictures of fibre cross-sections of these samples display a more even distribution of the dyestuff and a better penetration in the enzyme-treated wool fibres but the colour fastness of the enzyme-treated wool is decreased. Though the felting behaviour of the protease treated wool is significantly improved the felting tendency is still too high for an antifelting finish. An increased damage of the enzyme-treated wool in comparison with the untreated one was not observed. PMID- 11500222 TI - Enzymes and chelating agent in cotton pretreatment. AB - Desized cotton fabric and cotton seed-coat fragments (impurities) have been treated with commercial cellulase (Celluclast 1.5 L), hemicellulase-pectinase (Viscozyme 120 L) and xylanase (Pulpzyme HC) enzymes. Seed-coat fragments hydrolyzed much faster than the cotton fabric itself. This relative difference in hydrolysis rates makes possible a direct enzymatic removal of seed-coat fragments from desized cotton fabric. Addition of chelating agents such as ethylenediamine tetra-acetic acid (EDTA) markedly enhanced the directed enzyme action. Pretreatments carried out in acidic solution at pH 5 increased the lightness of seed-coat fragments, contrary to the samples treated in neutral medium at pH 7. Alkaline scouring resulted in darker seed-coat fragments except for the samples pretreated with Pulpzyme HC plus EDTA. This effect is similar to that observed in the biobleaching process in pulp and paper industry. PMID- 11500224 TI - The role of chitosan in wool finishing. AB - The application of the biopolymer chitosan as an alternative to the conventional contaminating processes in textiles was studied. As chitosan is produced by biomass, it is biodegradable and bioadsorbable. These properties are increasingly important given the current environmental legislation. The main aim of chitosan treatment of oxidised wool fabrics is to improve felting properties and dyeing behaviour. PMID- 11500225 TI - A study of the influence of structure on the effectiveness of chitosan as an anti felting treatment for wool. AB - The effect of chitosan on the resistance of wool fabric to felting on washing has been studied using nine structurally different samples of chitosan. Structural differences examined include molecular weight, level of N-acetylation, and the nature and concentration of homologous N-acyl groups. No strong dependency of shrinkage on molecular weight or level of N-acetylation has been found, but increasing the hydrophobic character of chitosan through the incorporation of a number of long-chain N-acyl groups gives improved anti-felting behaviour, compared to chitosan itself, at the same level of add-on. PMID- 11500227 TI - Potential and problems of developing transdermal patches for veterinary applications. AB - A new frontier in the administration of therapeutic drugs to veterinary species is transdermal drug delivery. The primary challenge in developing these systems is rooted in the wide differences in skin structure and function seen in species ranging from cats to cows. The efficacy of a transdermal system is primarily dependent upon the barrier properties of the targeted species skin, as well as the ratio of the area of the transdermal patch to the species total body mass needed to achieve effective systemic drug concentrations. A drug must have sufficient lipid solubility to traverse the epidermal barrier to be considered for delivery for this route. A number of insecticides have been developed in liquid "pour-on" formulations that illustrate the efficacy of this route of administration for veterinary species. The human transdermal fentanyl patch has been successfully used in cats and dogs for post-operative analgesia. The future development of transdermal drug delivery systems for veterinary species will be drug and species specific. With efficient experimental designs and available transdermal patch technology, there are no obvious hurdles to the development of effective systems in many veterinary species. PMID- 11500228 TI - Veterinary drug delivery: potential for skin penetration enhancement. AB - A range of topical products are used in veterinary medicine. The efficacy of many of these products has been enhanced by the addition of penetration enhancers. Evolution has led to not only a highly specialized skin in animals and humans, but also one whose anatomical structure and skin permeability differ between the various species. The skin provides an excellent barrier against the ingress of environmental contaminants, toxins, and microorganisms while performing a homeostatic role to permit terrestrial life. Over the past few years, major advances have been made in the field of transdermal drug delivery. An increasing number of drugs are being added to the list of therapeutic agents that can be delivered via the skin to the systemic circulation where clinically effective concentrations are reached. The therapeutic benefits of topically applied veterinary products is achieved in spite of the inherent protective functions of the stratum corneum (SC), one of which is to exclude foreign substances from entering the body. Much of the recent success in this field is attributable to the rapidly expanding knowledge of the SC barrier structure and function. The bilayer domains of the intercellular lipid matrices within the SC form an excellent penetration barrier, which must be breached if poorly penetrating drugs are to be administered at an appropriate rate. One generalized approach to overcoming the barrier properties of the skin for drugs and biomolecules is the incorporation of suitable vehicles or other chemical compounds into a transdermal delivery system. Indeed, the incorporation of such compounds has become more prevalent and is a growing trend in transdermal drug delivery. Substances that help promote drug diffusion through the SC and epidermis are referred to as penetration enhancers, accelerants, adjuvants, or sorption promoters. It is interesting to note that many pour-on and spot-on formulations used in veterinary medicine contain inert ingredients (e.g., alcohols, amides, ethers, glycols, and hydrocarbon oils) that will act as penetration enhancers. These substances have the potential to reduce the capacity for drug binding and interact with some components of the skin, thereby improving drug transport. However, their inclusion in veterinary products with a high-absorbed dose may result in adverse dermatological reactions (e.g., toxicological irritations) and concerns about tissue residues. These are important considerations when formulating a veterinary transdermal product when such compounds are added, either intentionally or otherwise, for their penetration enhancement ability. PMID- 11500229 TI - Aquaculture pharmaceuticals and biologicals: current perspectives and future possibilities. AB - With the continued expansion of cultured fin and shellfish species, aquaculture has become a key component of the animal health industry. The ever-increasing demand for quality pharmaceutical and biological products, to combat bacterial and viral infections, calls for the development of modern formulations and novel drug delivery systems. The purpose of this paper is to provide a general overview of important fish pathogens, manifested fish diseases, and control methods. Currently available pharmaceutical and biological products are tabulated. Emphasis is placed on the extension of formulation and process technologies, commonly encountered in human pharmaceuticals, to the aquaculture arena. New research efforts in the area of novel vaccine delivery systems, i.e. enteric coated beads and biodegradable microspheres, are also summarized. PMID- 11500230 TI - Bovine mastitis and intramammary drug delivery: review and perspectives. AB - Intramammary infections (IMIs) represent a major feature in bovine pathology. The treatment of IMIs concern antimicrobial substances. Therapeutic strategies involve administration of immediate release formulations during lactation with or without long-acting formulations during the dry period. Current treatments are not very successful and cure rates are poor, especially towards Staphylococcus aureus which is responsible for chronic infections and huge economic losses. New strategies have recently been investigated. These include particular immunomodulators like lysostaphin or cytokines, and novel formulations (e.g. liposomes, microparticles or nanoparticles) that allow uptake of the active component by phagocytes and thus prolong an enhanced antibacterial activity. PMID- 11500231 TI - Opportunities and obstacles in veterinary dental drug delivery. AB - Dentistry is a relatively new field in veterinary medicine, despite the fact that dental disease is common in dogs and cats. Obstacles to the development of dental treatments in animals include the diversity of the dentition among the species, the difficulty in administration of oral treatments, the different presentations of oral disease, and the cost of regulatory approval for each species. Mechanical removal of plaque and calculus has been the mainstay of periodontal disease treatment in animals and humans. New adjunctive therapies, as well as new applications for older drugs, are being introduced for periodontal therapy. PMID- 11500232 TI - Recent advances in bovine reproductive endocrinology and physiology and their impact on drug delivery system design for the control of the estrous cycle in cattle. AB - When methods of drug intervention are being developed to control estrous cycles, a thorough understanding of the endocrine and functional changes together with the reproductive behavior of the animals are essential. This review presents our current knowledge on reproductive endocrinology, physiology and behavior, and the methods of drug intervention to control estrous cycles. It also describes current efforts to develop advanced drug delivery systems that meet the animal scientist's demands to control the estrous cycle in cattle. PMID- 11500233 TI - Identification of two estrogen regulated genes associated with growth regulation of human breast cancer. AB - We have identified two estrogen regulated gene products in the E(2) growth inhibited human breast cancer xenograft, T61; one showing 100% homology to the human BAC clone RP11-112E16, the other 100% homology to the human CPR3/DNJ3 gene. Verification by Northern blot analyses showed an up-regulation of the BAC clone RP11-112E16 and the CPR3/DNJ3 mRNAs upon E(2) treatment. Treatment of T61 tumors with tamoxifen, leading to static tumor growth, also increased the expression of the BAC clone RP11-112E16 and the CPR3/DNJ3 mRNAs. A similar association between growth inhibition and BAC clone RP11-112E16 and CPR3/DNJ3 mRNA induction was observed in MCF-7 cells treated with ICI 182.780. In MCF-7 cells, treatment with E(2) resulted in growth stimulation concomitant with a decrease in the BAC clone RP11-112E16 and CPR3/DNJ3 mRNA expression. Treatment with a combination of E(2) and ICI 182.780 abolished the anti-estrogen induced increase in BAC clone RP11 112E16 and CPR3/DNJ3 mRNA expression, indicating that regulation of the gene products is mediated through the ER. The association between growth inhibition and BAC clone RP11-112E16 or CPR3/DNJ3 mRNA expression was supported by high expression of both gene products in brain tissue. Further investigations are ongoing to clarify the biological function of these two gene products. PMID- 11500234 TI - Absence of thyroid transcription factor-1 expression in human parathyroid and pituitary glands. AB - Thyroid transcription factor-1 (TTF-1), a tissue-specific nuclear transcription factor involved in the embryogenesis and differentiation of human thyroid, lung and brain, has been recently identified in other rat tissues, including parafollicular C cells and parathyroid chief cells. Based on this distribution, a possible role for this factor in calcium homeostasis has been suggested. This study investigated the presence of TTF-1 transcripts and protein in human tissues expressing the calcium sensing receptor (CaSR). Using a RT-PCR technique, complemented by Southern blot analysis, TTF-1 expression was detected in human C cells (two medullary thyroid carcinomas), but not in normal and adenomatous (four adenomas and three hyperplasia) parathyroid, and normal and adenomatous (six adenomas) pituitary tissues. CaSR was expressed in all samples. The absence of expression was confirmed by Western blot. In contrast to previous studies in the rat, this study demonstrates the absence of TTF-1 transcripts in the human adult parathyroid and pituitary glands, although a role for this factor during the ontogeny of these organs cannot be excluded. PMID- 11500235 TI - Estradiol-induced IP(3) mediates the estrogen receptor activity expressed in human cells. AB - The recent findings that estradiol-induced IP(3)/PKC-alpha signalling pathway triggers DNA synthesis in HepG2 cells, containing estrogen receptor unable to stimulate gene transactivation, raises the hypothesis that this pathway represents an alternative signalling present when the amount of estrogen receptor (ER) is insufficient to mediate genomic effects. beta-estradiol-stimulated DNA synthesis and target gene expression have been studied in HepG2 and, ER-alpha or ER-beta negative, HeLa cells. We also examined whether either receptor is required for rapid effects of estrogen on DNA synthesis. Finally, the consequences of increased ER expression on estrogen-induced DNA synthesis and synthetic target gene expression have been evaluated. Our data indicate that the E2-induced IP(3) production is dependent on expression of either ER-alpha or ER beta in both HepG2 and HeLa cells. Moreover, inhibition of the IP(3) second messenger pathway blocks E2-induced cellular actions suggesting that this second messenger is responsible for estrogen's rapid, non-genomic effects on both DNA synthesis and gene expression. PMID- 11500236 TI - Demonstration of estrogen receptor subtypes alpha and beta in human adipose tissue: influences of adipose cell differentiation and fat depot localization. AB - A novel ER-subtype, the ER-beta has recently been characterized in various tissues, furthermore five isoforms of the ER-beta are known (ER-beta1--ER-beta5). Using immunoblotting and real- time RT-PCR, ER-alpha and beta were studied in human adipose tissue. The expression of ER-alpha mRNA was equal in subcutaneous gluteal adipose tissue, subcutaneous abdominal and intra-abdominal adipose tissue, similar findings were obtained at the protein level. In contrast the amount of ER-beta1 (protein and mRNA) was significantly lower in intra-abdominal adipose tissue as compared with the subcutaneous adipose tissue (five-fold lower in women, P<0.005 and three-fold lower in men, P<0.005) whereas the expression of ER-beta4 and -beta5 mRNA isoforms were significantly higher in gluteal adipose tissue compared to subcutaneous abdominal adipose tissue. No significant gender differences in ER expression was detected in any of the fat depots investigated. During adipocyte differentiation the expression of ER-alpha, -beta4 and -beta5 mRNA declined, whereas, the expression of ER-beta1 mRNA was constant. In conclusion, the existence of ER-beta isoforms in human adipose tissue was demonstrated and the amount of these receptors was dependent upon fat depot localization, with much reduced expression of ER-beta1 in intra-abdominal adipose tissue compared to subcutaneous adipose tissue. These findings may indicate that estrogens could have differentiation and depot specific effects in human adipose tissue. PMID- 11500237 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin increases steady-state estrogen receptor-beta mRNA levels after CYP1A1 and CYP1B1 induction in rat granulosa cells in vitro. AB - Previous in-vitro investigations of rat granulosa cells (GC) have shown that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) inhibits estrogen secretion and FSH induced aromatase activity. Although TCDD exerted no effect on basal aromatase enzyme activity, TCDD did reduce steady-state aromatase mRNA levels in GC using competitive RT-PCR. TCDD is hypothesized to induce these changes through aromatic hydrocarbon receptor(AHR)-mediated gene transcription and the modulation of the estrogen receptor (ER)-signaling pathway. In this study we show that rat GC express mRNA for AHR and the AHR nuclear translocator (ARNT) as well as biomarkers of TCDD action, CYP1A1 and CYP1B1 mRNA. Basal CYP1A1 and ER-alpha mRNAs were present only in trace amounts. By relative RT-PCR analysis we showed that CYP1A1 and CYP1B1 mRNA were induced significantly by TCDD at 6 h and that induction of CYP1A1 was maintained throughout the experiment. Using competitive RT-PCR, we observed no significant change in the mRNA levels of ARNT between control and TCDD-treated GC. Both AHR and ER-beta mRNA levels increased significantly at 48 h with TCDD compared with controls. Since ER-beta mRNA was not increased significantly until 48 h in culture, we suggest that in rat GC, the observed ER-beta mRNA increase by TCDD might be a result of CYP1A1/CYP1B1 catalyzed estrogen metabolism and aromatase mRNA inhibition via AHR. PMID- 11500238 TI - Tilapia glycoprotein hormone alpha subunit: cDNA cloning and hypothalamic regulation. AB - The cDNA encoding the glycoprotein alpha (GPalpha) subunit of tilapia (Oreochromis mossambicus) was partially cloned using RACE-polymerase chain reaction (PCR) technique. The amplified cDNA was found to be 583 bases long, and to consist of a portion of the signal peptide, the full sequence encoding the mature peptide (94 amino acids) and the 3' untranslated region. Northern blot analysis revealed a single band of approximately 600 bp. Alignment of the deduced amino acids of the mature protein showed that the tilapia GPalpha subunit shares more than 80% identity with that of other perciform fish (i.e. striped bass, sea bream and yellowfin porgy) and less than 70% with that of more taxonomically remote fish and other vertebrates. Exposure of dispersed tilapia pituitary cells to salmon gonadotropin-releasing hormone (sGnRH) elevated GPalpha mRNA levels via both PKC and cAMP-protein kinase A (PKA) pathways. The transcript levels were also regulated by pituitary adenylate cyclase activating polypeptide (PACAP) and neuropeptide Y (NPY), both acting through PKC and PKA pathways. Moreover, a combined treatment of PACAP or NPY with GnRH seems to have an additive effect on the GPalpha subunit gene transcription. These results suggest that in tilapia the expression of GPalpha subunit is regulated by GnRH mainly via PKC and PKA pathways. Furthermore, PACAP and NPY can elevate the GnRH-stimulated GPalpha subunit transcription and can directly affect the subunit mRNA levels, via the same transduction pathways. PMID- 11500239 TI - Iodide organification defects resulting from cosegregation of mutated and null thyroid peroxidase alleles. AB - This report describes an intriguing combination of the thyroid peroxidase (TPO) alleles resulting in an iodide organification defect. Sequence analysis of the patient's TPO gene showed the presence of T-deletion in exon 14 of the TPO gene (T2512del). From the sequencing pattern, this new mutation of the TPO gene was thought to be homozygous. mRNA transfection studies in which mutated mRNA was transfected to CHO-K(1) cells by electroporation showed that the cells transfected with mutated mRNA expressed smaller TPO molecules than those of cells transfected with wild-type mRNA and that they had TPO activity. However, the smaller TPO molecules could not translocate onto the cell surface. To investigate T2512del in the parents, their genomic DNAs were sequenced. Results showed that the mother had T2512del but the father did not. However, when seven polymorphic positions reported earlier were analyzed, the mother showed two kinds of nucleotides at four positions but the patient and father showed only one nucleotide at all seven positions. We suspected a deletion of the TPO gene (2p25) in one of two second chromosomes, and analyzed the patient's chromosomes by FISH using TPO cDNA and N-myc genomic DNA as probes. N-myc genomic DNA exhibited two signals and TPO cDNA only one signal, although the G-band showed no morphological abnormalities. T2512-deleted and 2p25-deleted null alleles cosegregated from her parents, resulting in iodide organification defect in the patient. PMID- 11500240 TI - Hormonal control of ovarian cell production of insulin-like growth factor binding proteins. AB - To determine if the hormonal effects on insulin-like growth factor binding protein (IGFBP) production differed between granulosa and thecal cells, both cell types were collected and cultured in serum-free medium with various hormone treatments, arranged in three experiments. Following treatment, cells were enumerated and media were collected, concentrated 10-fold and subjected to ligand blotting. Experiment 1 revealed that > or =1.5 x 10(5) viable cells at plating were needed for maximal IGFBP production by granulosa and thecal cells. The major forms of IGFBPs produced were a 27-34-kDa IGFBP (IGFBP-2 and -5), and a 20-22-kDa IGFBP (IGFBP-4) by the granulosa cells and a 40-44-kDa IGFBP (IGFBP-3), 34-kDa IGFBP (IGFBP-2), 27-29-kDa IGFBP (IGFBP-5) and a 20-22-kDa IGFBP (IGFBP-4) by the thecal cells. In Experiment 2A, insulin stimulated production of IGFBP-5 by thecal cells, and basic fibroblast growth factor (bFGF) inhibited the insulin induced increase in IGFBP-5 production; epidermal growth factor (EGF) and luteinizing hormone were without effect. The small amounts of IGFBP-2 and -3 produced by thecal cells of Experiment 2A were not affected by treatment. Production of IGFBP-2/-5 by granulosa cells in Experiment 2B was inhibited by insulin, with EGF and bFGF further enhancing insulin's inhibitory effect; follicle-stimulating hormone was without effect. In Experiment 3A, insulin enhanced production of IGFBP-5 by thecal cells whereas glucagon blocked insulin's stimulatory effect. In contrast, insulin or glucagon alone had no effect on production of the IGFBP-4 by thecal cells but when combined inhibited IGFBP-4 production. The small amounts of IGFBP-2 and -3 produced by thecal cells of Experiment 3A were not affected by treatment. In Experiment 3B, production of IGFBP-2/-5 by granulosa cells was attenuated in the presence of cortisol with or without insulin and insulin plus glucagon; glucagon and cortisol decreased production of IGFBP-4 by granulosa cells. These results suggest that production of IGFBP-2, -4, and -5 by granulosa and thecal cells are differentially affected by hormonal stimuli, and that IGFBP-3 is more consistently produced by thecal cells than granulosa cells of cattle although its production was not hormonally regulated. PMID- 11500241 TI - Spatial and topological distribution of progesterone receptor A and B isoforms during human development. AB - Progesterone receptor (PR) is a member of the nuclear receptor superfamily. To date, two isoforms of PR have been identified, PR-A and PR-B. In progesterone responsive tissues, the relative ratio of PR-A and PR-B is considered to contribute to the tissue-specific actions of progesterone. In this study, we examined the distribution of PR-A and PR-B in human fetal tissues ranging from 11 to 40 gestational weeks using immunohistochemistry and RT-PCR analysis. PR immunoreactivity was detected in a wide range of fetal tissues until 20 weeks of gestation, but gradually decreased towards the late gestational period. However, PR continued to remain positive throughout the gestational period in the interstitial cells of Cajal and endocrine tissues. PR-B was demonstrated as the predominant isoform in comparison to PR-A in all fetal tissues examined. These findings suggest that progesterone may be involved in the development of fetal organs throughout the gestational period. PMID- 11500242 TI - Estrogen receptor beta dominates in baboon carotid after endothelial denudation injury. AB - Increasing evidence, mainly from rodents, suggests that the predominant estrogen receptor (ER) in arteries is the newly-described ERbeta. We have investigated the expression of the two ERs in baboon carotid artery before and after denudation injury. Prior to denudation, both full length receptors were detected in semiquantitative RT-PCR; in addition two ERalpha but no ERbeta splicing variants were found. After denudation, ERbeta mRNA increased five-fold and declined, whereas ERalpha mRNA expression remained low. Prior to and after denudation, two ERalpha-specific antibodies showed no reaction with the vessel wall. Instead, two affinity purified antisera to ERbeta demonstrated a weak but distinct reaction over vascular smooth muscle cells with predenudation specimens, escalating post denudation and declining thereafter. The results suggest that selective targeting to ERbeta should be attempted when designing estrogen-based vasculoprotective drug therapies devoid of uterotrophic side effects. PMID- 11500243 TI - Developmental expression, alternative splicing and gene copy number for the pituitary adenylate cyclase-activating polypeptide (PACAP) and growth hormone releasing hormone (GRF) gene in rainbow trout. AB - Both growth hormone-releasing hormone (GRF) and pituitary adenylate cyclase activating polypeptide (PACAP) are encoded on the same gene in fish, but not in mammals. Our objective was to examine the onset and pattern of expression for the grf/pacap gene and to determine whether there is more than one gene in rainbow trout. The results show that grf/pacap mRNA is first expressed at 4 days after fertilization and continues through to hatching. Alternative splicing at all developmental stages produces a full-length transcript and one lacking exon four, which encodes GRF. Thus, independent regulation of the hormones occurs throughout development. Southern analysis shows that two grf/pacap genes exist in trout, but only one gene is responsible for the two identified transcripts. Overexpression of the grf/pacap gene in transgenic fish was attempted, but did not succeed. We conclude that the early and continued expression of grf/pacap mRNA in trout embryos and regulation of the neuropeptide ratio suggests they have a role in early brain development apart from their later role in releasing pituitary hormones. PMID- 11500244 TI - A mathematical approach to predict the affinity of estrogen receptors alpha and beta binding to DNA. AB - Estrogen receptors alpha and beta (ERalpha and ERbeta) bind to specific DNA sequences, estrogen response elements (EREs), usually located in the promoters of estrogen-regulated genes. The consensus ERE contains two inverted repeats of the 5'-AGGTCA-3' half-site (1/2 ERE) separated by three base pairs (bp). Many estrogen-responsive gene promoters contain one or more direct repeats (DR) of 1/2 ERE. Here, we examined the affinity of ERalpha and ERbeta binding and estradiol (E(2))-induced transactivation from select EREs and DRs. The affinity of ERalpha and ERbeta binding to imperfect EREs in vitro can be predicted from equations using the number of 1/2 EREs and the number of (AT)-(GC) bp substitutions within the 15-bp candidate ERE sequence as independent variables. Transactivation by ERalpha and ERbeta correlates with the affinity of ER-ERE binding with the exception of ERalpha from two low-affinity EREs. The equations developed here can be used to screen the promoters of estrogen-responsive genes for candidate ERE sequences. PMID- 11500245 TI - Expression of neurotrophin receptors in rat testis. Upregulation of TrkA mRNA with hCG treatment. AB - We report the expression of TrkA, TrkB and TrkC mRNAs in adult rat testis. With in situ hybridisation a low signal for TrkB and TrkC could be seen in postmeiotic cells of the seminiferous epithelium, whereas no signal for TrkA could be observed in untreated animals. Animals treated with hCG showed an induction of TrkA mRNA in premeiotic cells 12 h after the treatment, whereas an injection with EDS had no effect on the expression of Trk mRNAs. With the RNAse protection assay a low signal for TrkA was seen in whole testis of hCG treated animals. In staged tubules low expression was seen at stages VII-XI of untreated animals. Animals injected with hCG revealed that TrkA induction was highest during stages VIIcd and VIII of the cycle. The distinct expression pattern of these high-affinity neurotrophin receptors suggests different roles for neurotrophins during spermatogenesis. Induction of TrkA mRNA by hCG suggests that high-affinity binding of NGF during stages VIIcd-VIII in premeiotic cells is under control of the hypothalamic-pituitary-testicular axis. PMID- 11500246 TI - Ecdysone-controlled mRNA stability in Drosophila salivary glands: deadenylation independent degradation of larval glue protein gene message during the larval/prepupal transition. AB - 20-Hydroxyecdysone induces poly(A) shortening and the subsequent degradation of transcripts encoding the larval glue protein LGP-1 in Drosophila virilis late third larval instar salivary glands. Degradation concurs with the transient increase of ribonucleolytic activities in the gland cells. In vitro nuclease assays using crude cytoplasmic extracts of ecdysone-treated salivary glands demonstrate degradation to be deadenylation-independent and that the induced ribonucleolytic activities initiate the degradation of the Lgp-1 transcripts in putative single-stranded loop regions. The independence of degradation from deadenylation is also found in vivo in transformed D. melanogaster carrying a modified Lgp-1 gene. PMID- 11500247 TI - Inhibitory effect of oxytocin on corticotrope function in humans: are vasopressin and oxytocin ying-yang neurohormones? AB - Oxytocin (OT) and vasopressin (VP) are very similar neurohypophyseal peptides. While VP is known as an ACTH stimulating factor synergistic to CRF since two decades, the inhibiting activity of OT, first demonstrated in the human, is now confirmed in various species including mouse and rat!It is likely that endogenous oxytocinergic system which can be activated by physiological and/or pharmacological manipulation can "buffer" the stress activated vasopressin-ACTH cortisol action. Since VP and OT share also opposite action on cognitive function, those two "sister" neuropeptides might be considered as "ago antagonist" or "ying-yang" neurohormones! PMID- 11500248 TI - Brief vs. long maternal separations in infancy: contrasting relationships with adult maternal behavior and lactation levels of aggression and anxiety. AB - We compared the effects of daily long (3 h), brief (15 min) or no maternal separation (LMS, BMS, NMS) on postnatal days 2-14 on maternal behavior, aggression and anxiety levels during lactation in adulthood. Animals subjected to LMS received less maternal grooming than animals subjected to BMS. Maternal behaviors, including nursing, pup-grooming (PG) frequency and proportion of total grooming (PG+self-grooming) and nest-building during the immediate postpartum period and on postpartum days 2 and 5 were lower in dams with LMS experience compared to dams with BMS experience. LMS dams attacked male rats placed in their home cages less quickly and less often than did BMS or NMS dams. LMS dams also exhibited more anxiety than BMS dams in the elevated plus maze test. Thus, maternal separation during the postnatal period (or associated changes in the amount of maternal grooming received) affected subsequent adult maternal behavior, aggression and anxiety. The mechanism for this remains to be discovered, however, it seems likely to involve alteration of the development of oxytocin receptors in the brain. PMID- 11500249 TI - Cortisol mediates redistribution of CD8+ but not of CD56+ cells after the psychological stress of public speaking. AB - The present study investigated the question if a pharmacological blockade of cortisol release with stress affects lymphocyte redistribution in healthy volunteers. It was expected that the well known increases in the number of CD8+ (T-suppressor/cytotoxic cells) and CD56+ (natural killer cells) after stress would not be downregulated in the absence of an appropriate cortisol response, since redistribution is markedly influenced by glucocorticoids. In a double blind design, forty healthy male volunteers were exposed to a brief psychological stressor (public speaking) and received a single oral dose of dexamethasone [DEX] (N=20) or placebo (N=20) the evening before the main experiment. Ratings on emotional states and blood samples for determination of hormones, CD8+, and CD56+ cell counts were obtained at different time points during the experiment. Stress of public speaking led to highly significant increases in catecholamine and cortisol concentrations, to subjective discomfort and, most pronounced, to high increases in the number of CD8+ and CD56+ cells. DEX neither influenced baseline levels of mood, catecholamines and cell numbers nor stress induced responses of mood and catecholamines. However, during the whole experiment cortisol concentrations were suppressed in the DEX-condition and the number of CD8+, but not CD56+, cells remained elevated at the end of the session, while in the placebo condition the numbers of these cells were decreased to baseline levels. The data demonstrate that cortisol seems to play an important role in stress induced redistribution patterns of CD8+ but not CD56+ cells. This, however, can be explained by different migration processes between those cells (e.g. different targets of migration) and, therefore, different glucocorticoid influences on target tissues. PMID- 11500250 TI - Reduced dopamine function in depressed patients is related to suicidal behavior but not its lethality. AB - Several lines of evidence suggest a role for dopamine in the control of suicidal behaviour. Previously, we suggested an involvement of D2-dopaminergic function in the biology of suicide by demonstrating a smaller growth hormone (GH) response to apomorphine, a dopaminergic agonist, in depressed patients who later died by suicide. The purpose of the present study was to assess GH response to apomorphine in major depressed in-patients with a history of highly lethal suicide attempt compared to depressed patients with a low lethal lifetime suicide attempt history and non-attempters. The study was performed in a sample of 26 male depressed in-patients with a history of suicide attempt compared to 26 male depressed non-attempters. We observed a significant difference between suicide attempters and non-attempters (for GH peak, 6.3+/-5.1 ng/ml vs 15.8+/-14.2 ng/ml, F=10.3, df=1, 50, P=0.002). Moreover, GH peak responses to apomorphine did not differ between depressed patients with a high lethal lifetime suicide attempt history and patients who made low lethal lifetime suicide attempt. In conclusion, the results of the present study support a role for dopamine in the biology of suicidal behaviour. More specifically, an impaired GH response to apomorphine could be a marker of suicide risk. PMID- 11500251 TI - Effects of acute progesterone administration in healthy postmenopausal women and normally-cycling women. AB - The goal of this study was to investigate the behavioral and subjective effects of single doses of progesterone (intramuscular) in post-menopausal women and in women with normal menstrual cycles. Although certain metabolites of progesterone (e.g., allopregnanolone) are known to bind to GABA(A) receptors and produce sedative-like effects in laboratory animals, few studies have examined the acute effects of these neurosteroids in humans. Postmenopausal women (N=10) received progesterone (25, 50, 100 mg im) or placebo at weekly intervals, and women with normal menstrual cycles (N=8) received progesterone (100 mg im) or placebo once a month during the early follicular phase. Dependent measures included plasma levels of progesterone and allopregnanolone, self-report measures of mood and subjective effects and psychomotor performance. Plasma concentrations of progesterone and allopregnanolone increased in a time and dose-dependent manner, with relatively little variability. The concentrations were similar in both groups, although the ratio of allopregnanolone to progesterone was higher in cycling women at certain time points. Contrary to expectations, progesterone produced only modest behavioral or subjective effects. In the cycling women, it produced mild sedative-like effects (i.e., decreases in ratings of Vigor and Friendliness). In the post-menopausal women, only the highest dose (100 mg) slightly increased ratings of feeling "sluggish". These results suggest that progesterone and its metabolites, at concentrations well beyond those attained during the normal menstrual cycle, produce only marginal sedative-like effects. These findings suggest that brief (i.e., several hours) increases in plasma levels of allopregnanolone do not have direct effects on mood. PMID- 11500252 TI - The relationship between stress induced cortisol levels and memory differs between men and women. AB - Epidemiological as well as experimental studies in elderly subjects have suggested that postmenopausal women are more susceptible to the memory impairing effects of elevated cortisol levels than elderly men. Little is known however about gender differences in the susceptibility to acute stress in young subjects. In the present study a total of 58 healthy young subjects learned a word list, with recall being tested after a brief distraction task. Twenty-two subjects had to learn the list after exposure to a psychosocial stressor (Trier Social Stress Test: TSST), while the remaining subjects served as controls. Free cortisol was determined via saliva samples taken before and 10 minutes after stress. Subjects exposed to the stressor, did not show impaired memory performance per se when compared to the control group. However the cortisol increase in response to the stressor was negatively correlated (r=-0.43, P<0.05) with the memory performance within the stressed group (i.e., subjects showing a larger cortisol response recalling less words than subjects showing only a small cortisol increase). Additional analysis revealed, that this correlation was solely caused by the strong association observed in men (r=-0.82, P<0.05), while no association was observed in women (r=-0.05, P=ns). Our data suggests, that gender modulates the association between cortisol and memory after stress. Whether these differences reflect activational effects of sex steroids or developmentally-programmed sex differences awaits to be determined. PMID- 11500253 TI - Pituitary LH reserve suggests high risk of bulimia in amenorrheic women. AB - The present study was aimed at investigating a) the risk of having bulimia in a heterogeneous population of secondary amenorrhea; b) the LH and FSH secretion under basal and stimulated conditions (GnRH challenge) according to the presence of bulimic risk in our study population; c) the clinical and endocrine factors predictive of the bulimic risk in amenorrheic women. Amenorrheic women (n=73; age: 23.1+/-4.8 yrs; BMI:20.2+/-2.2 kg/m2) filled in a self rating scale for bulimia (BITE) and were classified accordingly, as being at low risk (score <10), at medium risk (score between 10 and 24), and at high risk (score > or =25) of having bulimia. In each subject basal mean plasma LH levels were calculated over one hour, sampling every 10 minutes, while in a subgroup of 45 patients the area under the curve (AUC) of plasma LH and FSH levels following a challenge with two doses of GnRH (10+10 microg, every two hours), sampling every 15 minutes, was also evaluated. High risk of bulimia was present in 12.3% of the population whereas 45.2% showed a low risk and 42.5% were at medium risk of developing the disorder. Mann-Whitney U test revealed that basal LH values were differently distributed with significantly lower levels (P<0.046) in amenorrheic women at high risk of bulimia in comparison with amenorrheic women at low risk. The AUC of LH secretion following the first challenge of GnRH was significantly higher in amenorrheic women with a high risk of bulimia in respect with both groups of women at low (P<0.034) and medium (P<0.009) risk. A similar result was found with FSH AUC following the first GnRH challenge (P<0.04 high risk vs low risk and P<0.014 high risk vs medium risk). In a multiple regression analysis, the best model predicting the risk of bulimia (BITE total score) included both the LH response to GnRH challenge and BMI. In conclusion, when facing secondary amenorrhea at first consultation, long before a precise pathophysiologic diagnosis of the disease, low basal plasma LH levels and LH response to GnRH challenge may allow one to suspect the presence of abnormal eating pattern of bulimic type. PMID- 11500254 TI - The testosterone metabolite and neurosteroid 3alpha-androstanediol may mediate the effects of testosterone on conditioned place preference. AB - Testosterone (T) and pregnane neurosteroids can enhance conditioned place preference (CPP). The present experiment examined CPP produced by T and its androgenic metabolite dihydrotestosterone (DHT) and 3alpha-Androstanediol (3alpha diol; an androstane neurosteroid). Administration of 3alpha-diol (>DHT>T) to intact male Long-Evans rats, 1.0 mg daily for six days, 30 min prior to exposure to the non-preferred side of the CPP chamber significantly increased preference for the non-preferred side of the chamber compared to that seen in home cage controls. Levels of circulating 3alpha-diol were increased significantly in 3alpha-diol>DHT>T-administered rats, compared to rats that had vehicle administered or androgen-administration discontinued. Androgen administration decreased seminal vesicle weight and intrahypothalamic androgen receptor (AR) binding compared to that seen in rats that had vehicle administered or androgen administration discontinued. Testosterone, DHT, and 3alpha-diol decreased GABA stimulated chloride influx in cortical synaptoneurosomes, and muscimol binding in the hippocampus compared to that seen in rats with vehicle administered or that had androgen-administration discontinued. These data indicate that administration of 3alpha-diol is more effective at enhancing CPP and increasing circulating 3alpha-diol levels than is DHT or T administration, and that all of the androgen regimens employed decreased peripheral and hypothalamic androgen receptor binding and cortical and hippocampal GABA(A) receptor function. Hence, whether the effects of 3 alpha-diol on CPP are mediated by differential actions at ARs or GABA(A) receptors in particular brain regions needs to be determined. PMID- 11500255 TI - Impact of cortisol on buspirone stimulated prolactin release: a double-blind placebo-controlled study. AB - Buspirone is known to stimulate prolactin release. Clinical studies (e.g. in chronic fatigue syndrome) suggest that the response may be influenced by baseline cortisol levels. We conducted a double-blind placebo-controlled study to examine the relationship between the prolactin response to buspirone challenge and baseline cortisol level. Fifty healthy volunteers took part in the study. Buspirone was found to consistently elevate PRL levels above those seen following placebo administration. The PRL response as measured by area under the curve was highly correlated with the baseline cortisol level. PMID- 11500256 TI - Novel mechanisms and devices to enable successful transdermal drug delivery. AB - Optimisation of drug delivery through human skin is important in modern therapy. This review considers drug-vehicle interactions (drug or prodrug selection, chemical potential control, ion pairs, coacervates and eutectic systems) and the role of vesicles and particles (liposomes, transfersomes, ethosomes, niosomes). We can modify the stratum corneum by hydration and chemical enhancers, or bypass or remove this tissue via microneedles, ablation and follicular delivery. Electrically assisted methods (ultrasound, iontophoresis, electroporation, magnetophoresis, photomechanical waves) show considerable promise. Of particular interest is the synergy between chemical enhancers, ultrasound, iontophoresis and electroporation. PMID- 11500257 TI - A dynamic in vitro lipolysis model. I. Controlling the rate of lipolysis by continuous addition of calcium. AB - Lipolysis by pancreatic lipase was investigated with the aim to establish an in vitro lipolysis model, which can be used to investigate the dissolution of poorly soluble lipophilic drug substances at controlled hydrolysis rates. The effects of three experimental parameters -- the concentrations of bile salts and Ca(2+) and the lipase activity -- were investigated. The effect on the rate of hydrolysis of emulsified soybean oil was investigated in experiments in a pH-stat at pH 6.5 and 37 degrees C. The free fatty acids produced by the hydrolysis were titrated at pH 6.5. It was shown that all three investigated parameters influence the initial rate of hydrolysis, whereas only the lipase activity and the concentration of Ca(2+) affect the subsequent stages. It was also shown that the rate of lipolysis can be controlled by the rate of adding Ca(2+). Thus, it is possible to design an in vitro model using readily available and inexpensive materials in which the hydrolysis rate can be controlled by the continuous addition of Ca(2+). PMID- 11500258 TI - In vitro and in vivo evaluation of polyoxyethylene esters as dermal prodrugs of ketoprofen, naproxen and diclofenac. AB - Novel polyoxyethylene esters of ketoprofen (1(a-e)), naproxen (2(a-e)) and diclofenac (3(a-e)) were synthesized and evaluated as potential dermal prodrugs of naproxen, ketoprofen and diclofenac. These esters were obtained by coupling these drugs with polyoxyethylene glycols by a succinic acid spacer. The aqueous solubilities, lipophilicities and hydrolysis rates of esters 1(a-e), 2(a-e) and 3(a-e) were determined in a buffered solution and in porcine esterase. The permeation of these prodrugs through excised human skin was studied in vitro. Furthermore we investigated the in vivo topical anti-inflammatory activity of esters 1(d), 2(e) and 3(e), which showed the best in vitro profile, evaluating the ability of these compounds to inhibit methyl nicotinate (MN)-induced skin erythema on healthy human volunteers. Esters 1(a-e), 2(a-e) and 3(a-e) showed good water stability and rapid enzymatic cleavage and their hydrolysis rates, both chemical and enzymatic, were not significantly affected by the length of the polyoxyethylenic chain used as promoiety. Concerning in vitro percutaneous absorption studies, only esters 1(d-e), 2(d-e) and 3(c-e) showed an increased flux through stratum corneum and epidermis membranes compared to their respective parent drugs. In vivo results showed an interesting delayed and sustained activity of esters 1(d) and 3(e) compared to the parent drugs. In conclusion polyoxyethylene glycols could prove to be suitable promoieties for ketoprofen, naproxen and diclofenac design since esters 1(d-e), 2(d-e) and 3(c-e) showed some requirements (chemical stability, enzymatic lability and an increased skin permeation) needed to obtain successful dermal prodrugs. Furthermore, was observed an appreciable and sustained in vivo topical anti-inflammatory activity of esters 1(d) and 3(e), compared to the parent drugs, using MN-induced erythema in human volunteers as inflammation model. PMID- 11500259 TI - Effects of physical properties of powder particles on binder liquid requirement and agglomerate growth mechanisms in a high shear mixer. AB - A study was performed in order to elucidate the effects of the physical properties of small powder particles on binder liquid requirement and agglomerate growth mechanisms. Three grades of calcium carbonate having different particle size distribution, surface area, and particle shape but approximately the same median particle size (4-5 microm), were melt agglomerated with polyethylene glycol (PEG) 3000 or 20,000 in an 8-l high shear mixer at three impeller speeds. The binder liquid requirement was found to be very dependent on the packing properties of the powder, a denser packing resulting in a lower binder liquid requirement. The densification of the agglomerates in the high shear mixer could be approximately predicted by compressing a powder sample in a compaction simulator. With the PEG having the highest viscosity (PEG 20,000), the agglomerate formation and growth occurred primarily by the immersion mechanism, whereas PEG 3000 gave rise to agglomerate growth by coalescence. Powder particles with a rounded shape and a narrow size distribution resulted in breakage of agglomerates with PEG 3000, whereas no breakage was seen with PEG 20,000. Powder particles having an irregular shape and surface structure could be agglomerated with PEG 20,000, whereas agglomerate growth became uncontrollable with PEG 3000. When PEG 20,000 was added as a powder instead of flakes, the resultant agglomerates became rounder and the size distribution narrower. PMID- 11500260 TI - Investigation of a new mathematical model for compression of pharmaceutical powders. AB - A new compaction equation, the log-exp model: V=V(l)-w log(P)+V(e) exp(P/P(m)) is presented. The model presumes that two compaction processes: a logarithmic and an exponential decline may be active simultaneously. Using non-linear regression techniques the model gives an excellent fit to a number of model substances with wide differences in compaction behaviour. Compared to the Kawakita equation the model covers a broader range of the compaction profile. The new model and the Cooper and Eaton equation fit the data on the same level, but the parameters derived from the log-exp model seems to have more discriminative power between substances and have a close relation to the apparent plastic or brittle densification mechanism. The log-exp model has potential as a tool in estimation of the strength of agglomerated materials. A proposal for a set-up to an iterative non-linear regression calculation in a spreadsheet program is attached. PMID- 11500261 TI - First-pass of GTS-21 on canine gut wall and liver determined by portal-systemic concentration difference. AB - To clarify the cause of the canine individual variability of plasma concentration after oral administration of GTS-21 [(E)-3-(2,4-dimethoxybenzylidene)-3,4,5,6 tetra-hydro-2,3'-bipyridine dihydrochloride], we evaluated the absorption ratio (F(A)), intestinal availability (F(G)), and hepatic availability (F(H)). The bioavailability (F) was evaluated from the ratio of the area under the plasma concentration versus time curves after oral and intravenous administration. Three isoflurane anaesthetised dogs were fitted with an electromagnetic flow probe attached to the portal vein and cannulated through the portal and the femoral veins. After intraduodenal administration of GTS-21, both plasma concentrations were determined simultaneously. F(A) x F(G) was calculated from the portal systemic concentration difference taking into consideration the blood-plasma partition ratio. F(A) was calculated from the residual drug contents of the small intestine. F(H) was calculated by dividing F by F(A) x F(G). The F values were 0.072, 0.021, and 0.037, indicating an individual variability of ca. threefold. The F(A) values were close to 1, and the F(G) values ranged from 0.449 to 0.461. Accordingly, the F(H) values were estimated at 0.170, 0.047, and 0.083. GTS-21 was completely absorbed but lost by first-pass effects of passage through the gut wall and liver. The first-pass effect of liver is larger than that of the gut wall, and dominates the individual variability in plasma concentration. PMID- 11500262 TI - Lipophilicity and membrane interactions of cationic-amphiphilic compounds: syntheses and structure-property relationships. AB - This study was performed to elucidate the relationship between steric factors, lipophilicity, and the potency of cationic-amphiphilic compounds to displace calcium ions from phosphatidylserine monolayers. The latter property is considered to be a substance/phospholipid affinity measure. A series of cationic amphiphilic 3-phenyl-N,N-dimethylpropylamine derivatives with systematic structural variations was synthesized. Lipophilicity values were determined by chromatographic (RP-HPLC, log D(7.4)), shake-flask (log P), and theoretical (CLOGP) techniques. The potency of the compounds to displace calcium ions from phosphatidylserine monolayers was determined using a radiotracer technique, employing the isotope (45)Ca(2+). The experimental lipophilicity values of several isomeric biphenyl- and diphenyl-congeners differ more than could be expected from the CLOGP-calculations and show a good correlation to the calculated molecular surface areas. Although the affinity of the substances to the phospholipid monolayer tends to increase with lipophilicity, no general interrelation between the two properties could be found. Surprisingly, the assay system (a phospholipid monolayer) was quite sensitive towards small steric changes at the 'ligand' molecules. Stereochemical factors have a considerable influence on the interaction of solutes with phospholipid membranes. It must be questioned whether lipophilicity measures alone, without taking other molecular features into account, can meaningfully be used to explain or predict the influence of solutes on membrane-related processes and properties. PMID- 11500263 TI - Cancer epidemiology in the elderly. AB - Although age per se is not an important determinant of cancer risk, but rather a surrogate of prolonged carcinogen exposure, the increased life expectancy over the last few decades in most areas of the world has greatly increased cancer burden in the elderly. Trends in mortality for most common cancer sites in the elderly tended systematically to increase up to the early 1990s, but declined thereafter in North America and Western Europe in both sexes, and in Japanese females. Most trends remain unfavourable for elderly population of both sexes in eastern Europe, mainly reflecting patterns in smoking. Trends of incidence in elderly populations are more difficult to interpret, on account of changed diagnosis and certification for several cancer sites, including prostate cancer, brain tumours and multiple myeloma. Screening and early diagnosis for cancer in the elderly should be critically considered for prostate, but also for colorectal, breast and other selected neoplasms. PMID- 11500264 TI - The unexpected contribution of immunosenescence to the leveling off of cancer incidence and mortality in the oldest old. AB - In this paper the hypothesis that some features of immunosenescence might impact on the levelling off of cancer incidence and mortality in the oldest old will be considered. In fact, the term immunosenescence suggests that a progressive loss of immune system (IS) function occurs with aging. However, the age-related modifications of the IS can be more properly acknowledged as a 'remodeling' characterized by profound structural changes, which modify the functional properties of IS. We suggest that the expansion with age of natural killer cells (NK) and of T cells which progressively acquire phenotypes intermediate between T lymphocytes and NK cells, together with the age-related changes in the production of inflammatory/anti-inflammatory cytokines, such as INFgamma and IL-4, might create an environment unfavorable for neoplastic growth in the oldest old. In this perspective, studies on immunosenescence likely provide insights on mechanisms responsible for the individual capacity to escape from the life threatening consequences of cancer outgrowth. PMID- 11500265 TI - The elderly cancer patient: a nursing perspective. AB - Since cancer incidence tends to increase with age, health professionals will encounter ever-greater numbers of older people with cancer. Elderly cancer patients present complex problems that need comprehensive physical and psychosocial support. In order to give specialised care to this segment of the population, a multidisciplinary approach must be used; only in this way can an individualised treatment program be provided. Oncology nurses are an important component of this team and can contribute significantly to the panorama of needs of this segment of the population, which include the prevention and early detection of cancer, the use of state-of-the-art treatments, patient education, care during and after hospitalisation and quality of life (QOL) issues. In this way, the older person with cancer can be treated in an optimal manner and survival can hopefully be improved in a meaningful way. PMID- 11500266 TI - Home care and short-run nursing homes: organizational aspects of their integration with oncological organizations. AB - Social-health care to oncological elderly patients implies interconnection among oncological hospital and sub-district services and acknowledgement of a sole access channel. The project requires the formation of an inter-administrative coordination group and of functional transmural units with evaluational and operative roles. Various care levels (protected hospital admission and discharge, continuity visits, evaluational-therapeutic integration during treatment, palliative cures) implicate specific criterion of eligibility and actions to rationalize organization, coordination and distribution of interventions. Efficiency and effectiveness depend on integration with the services that supply material and with the diagnostic and ambulatory ones. The mid-term prospectives of the integration regard computerization of diagnostic, therapeutic, care and rehabilitation courses of patients (Regional Computerized Register of Disability) and formation of polyfunctional centres that concern home, residential and hospital intervention. Powerful technological instruments and the new organizational forms now available should encourage the formation of a morally upright society. PMID- 11500267 TI - Prevalence of functional disability among elderly patients with cancer. AB - This study is part of a larger multicenter prospective study conducted in Italy to assess the efficacy of the comprehensive geriatric assessment (CGA) among elderly patients with cancer (i.e. aged 65 years or older). The prevalence of functional limitations, and its association with selected characteristics, was investigated among 303 elderly patients consecutively admitted at the Department of Medical Oncology, IRCCS Centro di Riferimento Oncologico, Aviano (Northeast Italy), between 1995 and 1998. These patients had a median age of 72 years (range, 65-94), and were affected by haematological (n=182) or solid tumours (n=121). At baseline, their physical function was assessed, in addition to performance status (PS), by means of the activity of daily living (ADL) and the instrumental activities of daily living (IADL) scales. Overall, 17% of the patients had a limitation for ADL, and 59% for IADL, the prevalence of functional disabilities increased with age. Specifically, 8% of patients had continence limitations and 13% had limitations in taking the prescribed drugs. By multivariate analysis, a poor PS turned out to be a strong independent determinant of both ADL and IADL disabilities. PMID- 11500268 TI - Acute myeloid leukemia in the elderly: biology and therapeutic strategies. AB - Age represents one of the most important adverse prognostic factors in acute myeloid leukemia (AML). The therapeutic results for patients older than 60 years accrued into clinical trials of intensive chemotherapy are largely unsatisfactory (complete remission rates rarely superior to 50-60%; median relapse-free survival usually less than 12 months). Because only 30-40% of elderly patients are actually entered into these trials, the overall failure of current treatments appear even more disappointing when considered in the context of the whole population of older individuals with AML. This appears primarily due to intrinsic differences in the biology of leukemia itself and to host-related factors (i.e. reduced tolerance to chemotherapy and comorbidity). AMLs of older subjects display several biological overlaps with secondary AMLs including multilineage involvement, phenotype, unfavorable cytogenetics and elevated activity of multidrug resistance genes. The clinical application of biologically-based prognostic factors may enable to separate patients who may actually benefit from aggressive chemotherapy from those who should be offered attenuated/palliative treatments or enrolled upfront into experimental trials of new drugs or biologic/immunologic treatments. This may hopefully result in a 'risk-adapted' strategy aimed at improving disease free survival and/or quality of life for patients with differing risk profiles. PMID- 11500269 TI - Management of hematologic malignancies in the elderly: 15-year experience at the Aviano Cancer Center, Italy. AB - The optimal management of hemopoietic malignancies in the elderly requires specific strategies targeted at the peculiar clinico-biologic features of such patients. In the time frame spanning from 1983 to 1998, several clinical trials have been performed at the Aviano Cancer Center, Italy, aimed at extending the knowledge of natural history of hemopoietic malignancies in the elderly and evaluating the efficacy and toxicity of different chemotherapy programs specifically devised for older subjects affected by acute myeloid leukemia, myelodysplasia, non-Hodgkin's lymphomas, or multiple myeloma. Here the most relevant results are summarized and information stemmed from such studies. The present report rather than provide a comprehensive review on hematologic neoplasms in the elderly, mainly reflects the 'philosophy' deriving from 15 years of studies of the group in the field of geriatric oncology/hematology. Elaboration of a 'consensus' view will represent the challenge for all the investigators involved in such an important field of research. PMID- 11500270 TI - Aging and cancer: the geriatrician's point of view. PMID- 11500271 TI - The role of radiotherapy in the management of elderly cancer patients in light of the GROG experience. AB - 60-70% of all cancers will develop in individuals aged 65 and over during this century. Radiation therapy will play a major role in the treatment of cancer, especially in the elderly. The Italian Geriatric Oncology Group (GROG) in 1994 started a number of prospective studies to assess the characteristics of each elderly cancer patient referred to the radiotherapy centers (age, gender, performance status, comorbidity, ADL, family status) and the main features of the tumor in that patient. Based on the obtained results, the role of radiotherapy is underlined and future perspectives are identified. PMID- 11500272 TI - Developing departmental consensus in the search for a new chair. PMID- 11500273 TI - Call for a unified match. PMID- 11500275 TI - Biomedical scientists and physicians as experts in federal courts. PMID- 11500276 TI - From concept to culture: the WWAMI program at the University of Washington School of Medicine. AB - Shortages of primary care physicians have historically affected rural areas more severely than urban and suburban areas. In 1970, the University of Washington School of Medicine (UWSOM) administrators and faculty initiated a four-state, community-based program to increase the number of generalist physicians throughout a predominantly rural and underserved region in the U.S. Northwest. The program developed regional medical education for three neighboring states that lacked their own medical schools, and encouraged physicians in training to practice in the region. Now serving five Northwest states (Washington, Wyoming, Alaska, Montana, and Idaho), the WWAMI program has solidified and expanded throughout its 30-year history. Factors important to success include widespread participation in and ownership of the program by the participating physicians, faculty, institutions, legislatures, and associations; partnership among constituents; educational equivalency among training sites; and development of an educational continuum with recruitment and/or training at multiple levels, including K--12, undergraduate, graduate training, residency, and practice. The program's positive influences on the UWSOM have included historically early attention to primary care and community-based clinical training and development of an ethic of closely monitored innovation. The use of new information technologies promises to further expand the ability to organize and offer medical education in the WWAMI region. PMID- 11500277 TI - Lessons learned from a distance-based consulting program to assist faculty development projects. AB - Changes in faculty roles and demographics necessitate a re-examination of the types of professional development opportunities offered in academic institutions. A distance-based consulting program was designed to assist faculty development projects as they progress through all stages of faculty development: needs assessment, project design, implementation, and, in particular, program evaluation and dissemination of results (i.e., presentations and published articles). The progress of 17 faculty development projects in primary care educational sites that received assistance in the United States and Canada was tracked over two years. Three factors were identified as having the most impact on the success of faculty development projects: (1) funds committed to and designated for faculty development; (2) funded, protected time for at least one person to implement the faculty development initiative; and (3) an environment capable of supporting faculty development initiatives (e.g., no major budget shortfall, few faculty transitions, a strong mission, no threat of mergers). Only a few of the participating sites reached the stage of evaluating and publishing articles about the outcomes of their projects within the designated 15-month time frame, with many sites reporting environmental impediments to project success. The authors describe the institutional characteristics that facilitated project success, assess the usefulness of distance-based consulting efforts, and offer recommendations for future distance-based consulting programs. They conclude by noting that the personal touch (i.e., one-on-one contact with consultants) is what is most appreciated, and that excellent one-on-one, in-person assistance may be inherently more effective than even the best-run distance-based consulting. PMID- 11500278 TI - Should academic medical centers conduct clinical trials of the efficacy of intercessory prayer? AB - Intercessory prayers for health or healing are requests to an object of worship for the preservation or restoration of health. There has been a recent proliferation of clinical trials that compare the health outcome of a group of prayed-for patients with that of controls, to test the efficacy of intercessory prayer. In this essay, the author defines the concept of intercessory prayer, contrasts it with other forms of prayer, and reviews the literature concerning clinical trials of its efficacy. The arguments put forward in favor of conducting such trials and those against are described and the reader is invited to consider their relative merits. The author concludes by discussing the potential power of faith in healing, reviewing the philosophical basis and pitfalls of clinical trials of intercessory prayer, and urging readers to weigh the arguments for and against such trials in academic medicine. PMID- 11500279 TI - A process for reducing workload and enhancing residents' education at an academic medical center. AB - Academic medical centers are under increasing pressure to find alternatives to residents for the provision of patient care and to expand and improve the educational opportunities for residents. To address these concerns, the authors performed a study of the medical wards at Harborview Medical Center, a county owned medical center managed by the University of Washington School of Medicine. Admitting diagnoses, provider names, and billings were obtained from professional practice plan billing records. Based on the distribution of admitting diagnoses, a subset of patients was identified that could be removed from routine care by residents and could instead be cared for by non-physician providers (i.e., physician assistants and nurse practitioners) using clinical pathways. The cohort was large enough to reduce the number of patients per resident to within national accreditation guidelines, and to provide faculty with more time available for teaching. The authors summarize the approach used to identify the new model for care delivery indicated above and the plans made to implement that model and to analyze its impact on the quality of patient care, hospital costs, residents' education, and the process of implementing change. The authors conclude that solutions to the problems of workload and education that they confronted will vary by department and hospital setting. Yet a systematic approach to discovering solutions, such as they present, can be adapted to any setting. PMID- 11500280 TI - Who counts in medical school? PMID- 11500281 TI - What makes a good teacher? Lessons from teaching medical students. PMID- 11500282 TI - Coordination of cancer teaching 1951. PMID- 11500283 TI - Hospital of the transfiguration. PMID- 11500285 TI - Do clerkship experiences affect medical students' attitudes toward chronically ill patients? AB - PURPOSE: To measure changes in medical students' attitudes toward chronically ill patients, and to identify experiences, specifically during clerkships, that contributed to students' attitudes. METHOD: A cohort of students from five U.S. medical schools voluntarily participated in three surveys longitudinally administered before and after required clinical rotations. The first two questionnaires were identical and asked for demographic information and pre matriculation experiences with chronically ill patients. The third was modified to include questions about clinical experiences with chronically ill patients. Responses from the first and third questionnaires were linked for analysis. RESULTS: A total of 502 of 695 students (69%) completed both the first and the third questionnaires. Many students (36%) had had pre-matriculation experiences with chronic illness. After clinical training, 25% of the respondents stated that they would seek another career specialty if the incidence of chronically ill patients increased in their chosen field, compared with the 9% who responded so before clinical training (p <.001). While 73% of the students had favorable perceptions toward chronically ill patients, and 91% felt involved in care, significantly fewer students (p <.01) had had positive patient care experiences when working with residents (57%) and attendings (59%). Gender, age, prior experiences, and school site were not associated with attitudinal changes. CONCLUSION: Students begin medical school with positive attitudes toward caring for chronically ill patients, but this perception depreciates with clinical experience, which may affect specialty decisions. Contributing factors may include adequate role modeling by residents and attendings and a perceived discrepancy in the quality of care patients receive. PMID- 11500286 TI - Diminished use of osteopathic manipulative treatment and its impact on the uniqueness of the osteopathic profession. AB - PURPOSE: To determine whether osteopathic manipulative treatment (OMT), a key identifiable feature of osteopathic medicine, is becoming a "lost art" in the profession, and whether the long-term evolution of osteopathic medicine into mainstream medicine and particularly specialization has had a similar impact on the use of OMT by family practitioners and specialists. METHOD: In April 1998, a two-page questionnaire was mailed to 3,000 randomly selected osteopathic physicians in the United States to assess factors affecting their use of OMT. Descriptive statistics, linear regression analyses, and analysis of variance techniques were used to test for differences. RESULTS: The response rate was 33.2%. Over 50% of the responding osteopathic physicians used OMT on less than 5% of their patients, and analysis of variance revealed OMT use was significantly affected by practice type, graduation date, and family physicians versus specialists. For specialists, 58% of the variance regression was attributed to barriers to use, practice protocol, attitudes, and training, whereas for family physicians, 43% of the variance regression was attributed to barriers to use, practice protocol, and attitudes. More important, the eventual level of OMT use was related to whether postgraduate training had been undertaken in osteopathic, allopathic, or mixed staff facilities, particularly for osteopathic specialists. CONCLUSIONS: The evidence supports the assertion that OMT is becoming a lost art among osteopathic practitioners. Osteopathic as well as allopathic medical educators and policymakers should address the impact of the diminished use of OMT on both U.S. health care and the unique identifying practices associated with the osteopathic profession. PMID- 11500287 TI - Longitudinal trends in the applicant pool for U.S. medical schools, 1974--1999. AB - PURPOSE: To examine the pool of applicants to U.S. medical schools from 1974- 1999 for changes in size and demographics and to identify factors that may be associated with such changes. METHOD: Data on characteristics of the total applicant pools to U.S. medical schools for 1974--1999 were collected from the Association of American Medical Colleges' Data Warehouse. Data on undergraduates' receiving bachelor's degrees and unemployment rates were obtained from the National Center for Education Statistics and the U.S. Bureau of Labor Statistics, respectively. Variables such as race/ethnicity, gender, age, and first time or reapplicant status were compared across the study period. RESULTS: The percentage of women applicants increased from 20% of the pool in 1974 to 45% of the pool in 1999, while the percentage of men dropped from 80% to 55%. The number of underrepresented minority (URM) applicants increased 45% during the period, from 2,890 to 4,181, but URM applicants represented only 11% of the total applicant pool in 1999. Between 1974 and 1999, the number of URM men applying to medical school dropped by 18%, from 1,984 to 1,629, while the number of URM women nearly tripled, from 906 to 2,552. The number of Asian/Pacific Islander applicants increased dramatically, from 986 in 1974 to 7,622 in 1999, and they now represent a fifth of all applicants. The proportion of reapplicants grew when the applicant pool grew and it shrank when the applicant pool shrank. No relationship was found between the size of the applicant pool and economic indicators, age, or geographic origin. CONCLUSION: Changes in the proportions of women and Asian/Pacific Islander applicants were the driving force in the expansion of the applicant pool between 1974 and 1999. PMID- 11500288 TI - Administration of open-ended test questions by computer in a clerkship final examination. AB - PURPOSE: To evaluate the reliability, efficiency, and cost of administering open ended test questions by computer. METHODS: A total of 1,194 students in groups of approximately 30 were tested at the end of a required surgical clerkship from 1993 through 1998. For the academic years 1993--94 and 1994--95, the administration of open-ended test questions by computer was compared experimentally with administration by paper-and-pencil for two years. The paper and-pencil mode of the test was discontinued in 1995, and the administration of the test by computer was evaluated for all students through 1998. Computerized item analysis of responses was added to the students' post-examination review session in 1996. RESULTS: There was no significant difference in the performances of 440 students (1993--94 and 1994--95) on the different modes of test administration. Alpha reliability estimates were comparable. Most students preferred the computer administration, which the faculty judged to be efficient and cost-effective. The immediate availability of item-analysis data strengthened the post-examination review sessions. CONCLUSION: Routine administration of open ended test questions by computer is practical, and it enables faculty to provide feedback to students immediately after the examination. PMID- 11500289 TI - Using standardized patients as teachers: a concurrent controlled trial. AB - PURPOSE: To compare two methods of teaching physical assessment, a traditional faculty-taught course and a course with components taught by specially trained standardized patients (SPs), with respect to students' performances and costs. METHOD: Medical students in their second year and without preliminary course work in physical assessment were taught by faculty-led small groups. Students in their first year were taught by faculty-led lecture-demonstrations and exercises led by physical examination teaching associates (PETAs). Both groups of students were tested with a performance-based examination that involved six identical stations. The costs of both courses were calculated using faculty and SP salaries. RESULTS: There was no difference in students' performances on two of the stations, those involving the eye and abdominal examinations. The class that had been taught by PETAs, however, demonstrated a statistically significant performance advantage on the remaining four stations. The cost saving from using the PETAs was conservatively estimated at $24,155. CONCLUSION: Specially trained SPs can effectively teach the normal physical examination to medical students and are a less expensive alternative to traditional faculty small-group teaching methods. PMID- 11500290 TI - Effectiveness of a formal post-baccalaureate pre-medicine program for underrepresented minority students. AB - PURPOSE: To address the effectiveness of a formal postbaccalaureate (PB) experience for underrepresented minority (URM) students before medical school. The program provided an intense year-long experience of course work, research, and personal development. METHOD: There were 516 participants from one medical school: 15 URM medical students had completed the formal PB program, 58 students had done independent PB work before matriculation, and 443 students were traditional matriculants. Cognitive and academic indicators [college science and non-science grade-point averages (GPAs); biology, physics, and verbal MCAT scores; and percentage scores from first-year medical school courses] were compared for the three groups. RESULTS: Both groups of students with PB experience demonstrated competency in the first year of medical school consistent with traditional students even though the students who had completed the formal PB program had lower MCAT scores and lower college GPAs than did the traditional students. Traditional predictors of academic performance during the first year of medical school did not significantly contribute to actual academic performances of students from the formal PB program. CONCLUSION: The results support the use of a formal PB program to provide academic readiness and support for URM students prior to medical school. Such a program may also improve retention. Noncognitive variables, however, may be important to understanding the success of such students in medical school. PMID- 11500291 TI - Women on the editorial boards of major journals. AB - PURPOSE: To determine the percentages of women on the editorial boards of general and specialty medical journals in comparison with the numbers of women physicians in the journals' respective specialties. METHOD: The numbers of women editors, deputy editors, assistant editors, and members of editorial boards of 12 major journals in 1999 were counted and compared with the percentages of women physicians in the journals' specialties, as published by the American Medical Association. RESULTS: Parity between the percentages of women on editorial boards of specialty journals and women physicians in the journals' specialties was found for five journals. Only one journal had more women on the editorial board than there were women physicians in the specialty. CONCLUSIONS: Fewer than half of the journals studied had parity between the percentages of women members of editorial boards and the percentages of women physicians in the specialties. Parity should be maintained to accurately reflect the numbers of women physicians in these fields. PMID- 11500292 TI - Bringing the human genome and the revolution in bioinformatics to the medical school classroom: a case report from Washington University School of Medicine. AB - The human genome project is revolutionizing medical research and the practice of clinical medicine. To understand and participate in this revolution, physicians must be fluent in human genomics and bioinformatics. At Washington University School of Medicine (WUSM), the authors designed a module for teaching these skills to first-year students. The module uses clinical cases as a platform for accessing information stored in GenBank, Online Mendelian Inheritance in Man (OMIM), and PubMed databases at the National Center for Biotechnology Information (NCBI). This module, which is also designed to reinforce problem-solving skills, has been integrated into WUSM's first-year medical genetics course. PMID- 11500293 TI - Widening the lens on standardized patient assessment: what the encounter can reveal about the development of clinical competence. AB - The standardized patient (SP) examination is used in a majority of medical schools to test clinical skills. This examination usually yields both numerical ratings of clinical skill and narrative comments by patients or observers, yet most empirical studies of SP assessment focus on the numerical ratings only. This quantitative focus can lead to a narrow conceptualization of the nature and development of clinical competence. The authors suggest that in addition to utilizing SP numerical ratings, medical educators also use the rich qualitative material produced in the SP examination (e.g., patient comments, videotapes of the examination) to explore students' development of clinical competence, which involves the purposive integration of basic science, technical skill, empathy, communication, professional role, and personal history. PMID- 11500294 TI - An inexpensive patient-encounter log. AB - Many medical schools use patient-encounter log forms to document and improve training experiences; however, they are expensive and/or time-consuming to analyze. The patient-encounter log form developed at the University of Hawaii is a one-page form on which students in the family medicine clerkship are required to maintain a daily record of patient demographics and diagnoses. The form is reviewed throughout the clerkship, and efforts are made to ensure that students' learning is supplemented to cover any gaps discovered with the log forms. The simple paper log form can be visually analyzed in less than a minute. During the initial implementation period, 44 students completed the forms. Twelve of these students also returned an end-of-year survey, which indicated that updating the logs required less than one minute per patient. The information obtained from the log forms was used to document and improve the consistency of the educational experience and to establish clinical curriculum guidelines for the clerkship. This form can easily be reproduced and adapted to other settings. PMID- 11500295 TI - Methods for measuring glycogen cycling. AB - Simultaneous synthesis and breakdown of glycogen is called glycogen cycling. The extent of hyperglycemia and decreased glycogen stores in diabetes mellitus may relate in part to the extent cycling occurs. Four methods have been introduced to estimate its extent in liver in humans. 1) In the fasted state, the rate of net hepatic glycogenolysis, i.e., glycogen breakdown minus synthesis, is estimated using NMR, and the rate of glycogenolysis is estimated from deuterium labeling of blood glucose on (2)H(2)O ingestion. 2) The rate of glycogen synthesis is estimated from the rate of labeling of carbon 1 of glycogen on [1-(13)C]glucose infusion, monitored by NMR, and the rate of breakdown from the rate of disappearance of that labeling on unlabeled glucose infusion. 3) The rate of synthesis from glucose-1-P, formed by glycogenolysis, is measured by the decrease in the (3)H/(14)C ratio in acetaminophen glucuronide on acetaminophen and [2 (3)H,6-(14)C]galactose administration. 4) The rate of synthesis is estimated from the dilution of label from labeled galactose in its conversion to the acetaminophen glucuronide, and the rate of glycogenolysis is estimated from the amount of label in blood glucose. In the first method, the fate of glucose-6-P is assumed to be only to glycogen and glucose. In the second, only glucose-6-P molecules formed by breakdown that are not cycled back to glycogen are measured. In the third, (3)H is assumed to be removed completely during cycling, and only the molecules cycled back to glycogen are measured. In the fourth, galactose conversion to glucose is assumed to be via glycogen. Quantitations in all four methods depend on assuming the order in which the molecules deposited in glycogen are released. PMID- 11500296 TI - Destabilization of TNF-alpha mRNA by retinoic acid in hepatic macrophages: implications for alcoholic liver disease. AB - Retinoic acid (RA) inhibits hepatic macrophage (HM) cytokine expression, and retinoids are depleted in alcoholic liver disease (ALD). However, neither the causal link between the two nor the mechanism underlying RA-mediated HM inhibition is known. The aim of the present study was to determine the mechanism of RA-induced inhibition of HM tumor necrosis factor (TNF)-alpha expression and the relevance of this regulation to ALD. Treatment with all-trans RA (500 nM) caused a 50% inhibition in lipopolysaccharide (LPS)-stimulated TNF-alpha expression by cultured normal rat HM. The mRNA levels for inducible nitric oxide synthase, interleukin (IL)-6, IL-1alpha, and IL-1beta were also reduced, whereas those for transforming growth factor-beta1, MMP-9, and membrane cofactor protein 1 were unaffected. The inhibitory effect on TNF-alpha expression was reproduced by LG268, a retinoid X receptor (RXR)-specific ligand, but not by TTNPB, an RA receptor (RAR)-specific ligand. RA did not alter LPS-stimulated NF-kB and activation protein-1 binding but significantly decreased TNF-alpha mRNA stability in HM. HM isolated from the ALD model showed significant decreases in all-trans RA (-48%) and 9-cis RA (-61%) contents, RA response element (RARE) binding, and mRNA levels for RARbeta, RXRalpha, and cytosolic retinol binding protein-1, whereas TNF-alpha mRNA expression was induced. TNF-alpha mRNA stability was increased in these cells, and an ex vivo treatment with all-trans RA normalized both RARbeta and TNF-alpha mRNA levels. These results demonstrate the RA-induced destabilization of TNF-alpha mRNA by cultured HM and the association of RA depletion with increased TNF-alpha mRNA stability in HM from experimental ALD. These findings suggest that RA depletion primes HM for proinflammatory cytokine expression in ALD, at least in part, via posttranscriptional regulation. PMID- 11500297 TI - Deficiency of dietary EAA preferentially inhibits mRNA translation of ribosomal proteins in liver of meal-fed rats. AB - The goal of these studies was to investigate the mechanisms by which amino acid supply regulates global rates of protein synthesis as well as the translation of ribosomal protein (rp) mRNAs in liver. In the experiments conducted, male weanling rats were trained over a 2-wk period to consume their daily food intake within 3 h. On day 14, rats were fed the control diet or an isocaloric, isonitrogenous diet lacking glycine, tryptophan, leucine, or the branched-chain amino acids (BCAA) for 1 h. Feeding Trp-, Leu-, or BCAA-deficient diets resulted in significant reductions in serum insulin, hepatic protein synthesis, eukaryotic initiation factor 2B (eIF2B) activity, and phosphorylation of eIF4E-binding protein 1 (4E-BP1) and ribosomal protein S6 kinase (S6K1). Phosphorylation of eIF2alpha was inversely related to eIF2B activity under all conditions. Alterations in the hepatic synthesis of rp were assessed by changes in the distribution of rp (S4, S8, L26) mRNAs across sucrose density gradients and compared with non-rp (beta-actin, albumin) mRNAs. In all dietary treatments, non rp mRNAs were mostly polysome associated. Conversely, the proportion of rp mRNAs residing in polysomes was two- to fivefold less in rats fed diets lacking tryptophan, leucine, or BCAA compared with rats fed the control diet. Total hepatic abundance of all mRNAs examined did not differ among treatment groups. For all parameters examined, there were no differences between rats fed the glycine-deficient diet and rats fed the control diet. The data suggest that essential amino acid (EAA) deficiency inhibits global rates of liver protein synthesis via a block in translation initiation. Additionally, the translation of rp mRNAs is preferentially repressed in association with decreased S6K1 phosphorylation. PMID- 11500298 TI - Sulfate transport in porcine thyroid cells. Effects of thyrotropin and iodide. AB - In porcine thyroid cells, thyroglobulin sulfation is controlled by thyrotropin (TSH) and iodide, which contribute to regulating the intracellular sulfate concentration, as we previously established. Here, we studied the transport of sulfate and its regulation by these two effectors. Kinetic studies were performed after [(35)S]sulfate was added to either the basal or apical medium of cell monolayers cultured without any effectors, or with TSH with or without iodide. The basolateral uptake rates were about tenfold higher than the apical uptake rates. TSH increased the basolateral and apical uptake values (by 24 and 9%, respectively, compared with unstimulated cells), and iodide inhibited these effects of TSH. On the basis of results of the pulse-chase experiments, the basolateral and apical effluxes appeared to be well balanced in unstimulated cells and in cells stimulated by both TSH and iodide: approximately 40-50% of the intracellular radioactivity was released into each medium, whereas in the absence of iodide, 70% of the intracellular radioactivity was released on the basolateral side. The rates of transepithelial sulfate transport were increased by TSH compared with unstimulated cells, and these effects decreased in response to iodide. These results suggest that TSH and iodide may each control the sulfate transport process on two sides of the polarized cells, and that the absence of iodide in the TSH-stimulated cells probably results in an unbalanced state of sulfate transport. PMID- 11500299 TI - Catecholamines inhibit Ca(2+)-dependent proteolysis in rat skeletal muscle through beta(2)-adrenoceptors and cAMP. AB - Overall proteolysis and the activity of skeletal muscle proteolytic systems were investigated in rats 1, 2, or 4 days after adrenodemedullation. Adrenodemedullation reduced plasma epinephrine by 95% and norepinephrine by 35% but did not affect muscle norepinephrine content. In soleus and extensor digitorum longus (EDL) muscles, rates of overall proteolysis increased by 15-20% by 2 days after surgery but returned to normal levels after 4 days. The rise in rates of protein degradation was accompanied by an increased activity of Ca(2+) dependent proteolysis in both muscles, with no significant change in the activity of lysosomal and ATP-dependent proteolytic systems. In vitro rates of Ca(2+) dependent proteolysis in soleus and EDL from normal rats decreased by ~35% in the presence of either 10(-5) M clenbuterol, a beta(2)-adrenergic agonist, or epinephrine or norepinephrine. In the presence of dibutyryl cAMP, proteolysis was reduced by 62% in soleus and 34% in EDL. The data suggest that catecholamines secreted by the adrenal medulla exert an inhibitory control of Ca(2+)-dependent proteolysis in rat skeletal muscle, mediated by beta(2)-adrenoceptors, with the participation of a cAMP-dependent pathway. PMID- 11500300 TI - Effects of antecedent hypoglycemia, hyperinsulinemia, and excess corticosterone on hypoglycemic counterregulation. AB - This study aimed to differentiate the effects of repeated antecedent hypoglycemia, antecedent marked hyperinsulinemia, and antecedent increases in corticosterone on counterregulation to subsequent hypoglycemia in normal rats. Specifically, we examined whether exposure to hyperinsulinemia or elevated corticosterone per se could impair subsequent counterregulation. Four groups of male Sprague-Dawley rats were used: 1) normal controls (N) had 4 days of sham antecedent treatment; 2) an antecedent hypoglycemia group (AH) had 7 episodes of hyperinsulinemic hypoglycemia over 4 days; 3) an antecedent hyperinsulinemia group (AE) had 7 episodes of hyperinsulinemic euglycemia; and 4) an antecedent corticosterone group (AC) had 7 episodes of intravenous corticosterone to simulate the hypoglycemic corticosterone levels in AH rats. On day 5, hyperinsulinemic euglycemic-hypoglycemic clamps were performed. Epinephrine responses to hypoglycemia were impaired (P < 0.05 vs. N) after antecedent hypoglycemia and hyperinsulinemia. This correlated with diminished (P < 0.05 vs. N) absolute glucose production responses in AH rats and diminished incremental glucose production responses in AE rats. Paradoxically, norepinephrine responses were increased (P < 0.05 vs. N) after antecedent hypoglycemia. Glucagon and corticosterone responses were unaffected by antecedent hypoglycemia and hyperinsulinemia. In AC rats, incremental but not absolute glucose production responses were decreased (P < 0.05 vs. N). However, neuroendocrine counterregulation was unaltered. We conclude that both antecedent hypoglycemia and hyperinsulinemia impair epinephrine and glucose production responses to subsequent hypoglycemia, suggesting that severe recurrent hyperinsulinemia may contribute to the development of hypoglycemia-associated autonomic failure. PMID- 11500301 TI - Leucine and insulin activate p70 S6 kinase through different pathways in human skeletal muscle. AB - Amino acids and insulin have anabolic effects in skeletal muscle, but the mechanisms are poorly understood. To test the hypothesis that leucine and insulin stimulate translation initiation in human skeletal muscle by phosphorylating 70 kDa ribosomal protein S6 kinase (p70(S6k)), we infused healthy adults with leucine alone (n = 6), insulin alone (n = 6), or both leucine and insulin (n = 6) for 2 h. p70(S6k) and protein kinase B (PKB) serine(473) phosphorylation were measured in vastus lateralis muscles. Plasma leucine increased from approximately 116 to 343 micromol/l during the leucine-alone and leucine + insulin infusions. Plasma insulin increased to approximately 400 pmol/l during the insulin-alone and leucine + insulin infusions and was unchanged during the leucine-alone infusion. Phosphorylation of p70(S6k) increased 4-fold in response to leucine alone, 8-fold in response to insulin alone, and 18-fold after the leucine + insulin infusion. Insulin-alone and leucine + insulin infusions increased PKB phosphorylation, but leucine alone had no effect. These results show that physiological concentrations of leucine and insulin activate a key mediator of protein synthesis in human skeletal muscle. They suggest that leucine stimulates protein synthesis through a nutrient signaling mechanism independent of insulin, raising the possibility that administration of branched-chain amino acids may improve protein synthesis in insulin-resistant states. PMID- 11500302 TI - Amino acids do not suppress proteolysis in premature neonates. AB - To determine whether increased amino acid availability can reduce proteolysis in premature neonates and to assess the capacity of infants born prematurely to acutely increase the irreversible catabolism of the essential amino acids leucine (via oxidation) and phenylalanine (via hydroxylation to form tyrosine), leucine and phenylalanine kinetics were measured under basal conditions and in response to a graded infusion of intravenous amino acids (1.2 and 2.4 g. kg(-1). day(-1)) in clinically stable premature (approximately 32 wk gestation) infants in the 1st wk of life. In contrast to the dose-dependent suppression of proteolysis seen in healthy full-term neonates, the endogenous rates of appearance of leucine and phenylalanine (reflecting proteolysis) were unchanged in response to amino acids (297 +/- 21, 283 +/- 19, and 284 +/- 31 micromol. kg(-1). h(-1) for leucine and 92 +/- 6, 92 +/- 4, and 84 +/- 7 micromol. kg(-1). h(-1) for phenylalanine). Similar to full-term neonates, leucine oxidation (40 +/- 5, 65 +/- 6, and 99 +/- 7 micromol. kg(-1). h(-1)) and phenylalanine hydroxylation (12 +/- 1, 16 +/- 1, and 20 +/- 2 micromol. kg(-1). h(-1)) increased in a stepwise fashion in response to graded amino acids. This capacity to increase phenylalanine hydroxylation may be crucial to meet tyrosine needs when exogenous supply is limited. Finally, to determine whether amino acids stimulate glucose production in premature neonates, glucose rate of appearance was measured during each study period. In response to amino acid infusion, rates of endogenous glucose production were unchanged (and near zero). PMID- 11500303 TI - ATP from glycolysis is required for normal sodium homeostasis in resting fast twitch rodent skeletal muscle. AB - Myocellular sodium homeostasis is commonly disrupted during critical illness for unknown reasons. Recent data suggest that changes in intracellular sodium content and the amount of ATP provided by glycolysis are closely related. The role of glycolysis and oxidative phosphorylation in providing fuel to the Na(+)-K(+) pump was investigated in resting rat extensor digitorum longus muscles incubated at 30 degrees C for 1 h. Oxidative inhibition with carbonyl cyanide m chlorophenylhydrazone, known as CCCP (0.2 microM), or by hypooxygenation did not alter myocellular sodium or potassium content ([Na(+)](i), [K(+)](i), respectively), whereas treatment with iodoacetic acid (0.3 mM), which effectively blocked glycolysis, dramatically increased [Na(+)](i) and the [Na(+)](i)/[K(+)](i) ratio. Experiments using ouabain and measurements of myocellular high-energy phosphates indicate that Na(+)-K(+)-ATPase activity is only impaired when glycolysis is inhibited. The data suggest that normal glycolysis is required to regulate intracellular sodium in fast-twitch skeletal muscles, because it is the predominant source of the fuel for the Na(+)-K(+) ATPase. PMID- 11500305 TI - The catecholamine response to spaceflight: role of diet and gender. AB - Compared with men, women appear to have a decreased sympathetic nervous system (SNS) response to stress. The two manifestations where the sexual dimorphism has been the most pronounced involve the response of the SNS to fluid shifts and fuel metabolism during exercise. The objectives of this study were to investigate whether a similar sexual dimorphism was found in the response to spaceflight. To do so, we compared catecholamine excretion by male and female astronauts from two similar shuttle missions, Spacelab Life Sciences 1 (SLS1, 1991) and 2 (SLS2, 1993) for evidence of sexual dimorphism. To evaluate the variability of the catecholamine response in men, we compared catecholamine excretion from the two SLS missions against the 1996 Life and Microgravity Sciences Mission (LMS) and the 1973 Skylab missions. RESULTS: No gender- or mission-dependent changes were found with epinephrine. Separating out the SLS1/2 data by gender shows that norepinephrine excretion was essentially unchanged with spaceflight in women (98 +/- 10%; n = 3) and substantially decreased with the men (41 +/- 9%; n = 4, P < 0.05). Data are a percentage of mean preflight value +/- SE. Comparisons among males demonstrated significant mission effects on norepinephrine excretion. After flight, there was a transient increase in norepinephrine but no evidence of any gender-specific effects. We conclude that norepinephrine excretion during spaceflight is both mission and gender dependent. Men show the greater response, with at least three factors being involved, a response to microgravity, energy balance, and the ratio of carbohydrate to fat in the diet. PMID- 11500304 TI - Induction of osteoblast differentiation indexes by PTHrP in MG-63 cells involves multiple signaling pathways. AB - Parathyroid hormone (PTH)-related peptide (PTHrP) can modulate the proliferation and differentiation of a number of cell types including osteoblasts. PTHrP can activate a G protein-coupled PTH/PTHrP receptor, which can interface with several second-messenger systems. In the current study, we have examined the signaling pathways involved in stimulated type I collagen and alkaline phosphatase expression in the human osteoblast-derived osteosarcoma cells, MG-63. By use of Northern blotting and histochemical analysis, maximum induction of these two markers of osteoblast differentiation occurred after 8 h of treatment with 100 nM PTHrP-(1-34). Chemical inhibitors of adenylate cyclase (H-89) or of protein kinase C (chelerythrine chloride) each diminished PTHrP-mediated type I collagen and alkaline phosphatase stimulation in a dose-dependent manner. These effects of PTHrP could also be blocked by inhibiting the Ras-mitogen-activated protein kinase (MAPK) pathway with a Ras farnesylation inhibitor, B1086, or with a MAPK inhibitor, PD-98059. Transient transfection of MG-63 cells with a mutant form of Galpha, which can sequester betagamma-subunits, showed significant downregulation of PTHrP-stimulated type I collagen expression, as did inhibition of phosphatidylinositol 3-kinase (PI 3-kinase) by wortmannin. Consequently, the betagamma-PI 3-kinase pathway may be involved in PTHrP stimulation of Ras. Collectively, these results demonstrate that, acting via its G protein-coupled receptor, PTHrP can induce indexes of osteoblast differentiation by utilizing multiple, perhaps parallel, signaling pathways. PMID- 11500306 TI - Metabolism of rabbit plasma-derived factor VII in relation to prothrombin in rabbits. AB - In the human circulation, factor VII is present in relatively low plasma concentration (0.01 microM) and has been reported to have a short half-life (t(1/2); 6 h). In contrast, prothrombin is present in a relatively high plasma concentration (2 microM) and has a relatively long catabolic half-life (t(1/2) = approximately 2-3 days). This report examines the metabolic characteristics of purified rabbit plasma factor VII and prothrombin, radiolabeled with (125)I and (131)I, respectively, in healthy young rabbits. From the plasma clearance curves of protein-bound radioactivities, fractional catabolic rates and compartmental distributions were calculated using a three-compartment model. Turnover of factor VII within the intravascular space (2.95 days) exceeded that of prothrombin (1.9 days). However, the whole body fractional catabolic rate of factor VII (0.34 days(-1); catabolic t(1/2) = 2.04 days) was significantly slower than that of prothrombin (0.53 days(-1); t(1/2) = 1.31 days). Furthermore, the fractional distributions of factor VII in the intravascular (0.14) and extravascular compartments (0.76) differed from those of prothrombin (0.29 and 0.53). Absolute quantities of factor VII and prothrombin catabolized by a 3-kg rabbit amounted to 0.18 and 24.0 mg/day, respectively (molar ratio of prothrombin to factor VII = 100). The molar ratio of catabolism was compared with the release rates of factor VII and prothrombin from rabbit livers perfused ex vivo. After correction for uptake of factor VII and prothrombin by the liver, the molar ratio of released prothrombin to factor VII in the perfusate was approximately 293:1 over a 0.25- to 3-h interval. These results indicate that, compared with prothrombin, factor VII in the healthy rabbit circulates as a relatively long-lived protein. This behavior does not reflect that reported for factor VII in the human circulation. PMID- 11500307 TI - Attenuation of age-related declines in glucagon-mediated signal transduction in rat liver by exercise training. AB - This study investigated alterations in glucagon receptor-mediated signal transduction in rat livers from 7- to 25-mo-old animals and examined the effects of exercise training on ameliorating these changes. Sixty-six young (4 mo), middle-aged (12 mo), and old (22 mo) male Fischer 344 rats were divided into sedentary and trained (treadmill running) groups. Isolated hepatic membranes were combined with [(125)I-Tyr(10)]monoiodoglucagon and nine concentrations of glucagon to determine maximal binding capacity (B(max)) and dissociation constant (K(d)). No alterations were found in B(max) among groups; however, middle-aged trained animals had significantly higher glucagon affinity (lower K(d); 21.1 +/- 1.8 nM) than did their untrained counterparts (50.2 +/- 7.1 nM). Second messenger studies were performed by measuring adenylyl cyclase (AC) specific activity under basal conditions and with four pharmacological stimulations to assess changes in receptor-dependent, G protein-dependent, and AC catalyst-dependent cAMP production. Age-related declines were observed in the old animals under all five conditions. Training resulted in increased cAMP production in the old animals when AC was directly stimulated by forskolin. Stimulatory G protein (G(s)) content was reduced with age in the sedentary group; however, training offset this decline. We conclude that age-related declines in glucagon signaling capacity and responsiveness may be attributed, in part, to declines in intrinsic AC activity and changes in G protein [inhibitory G protein (G(i))/G(s)] ratios. These age-related changes occur in the absence of alterations in glucagon receptor content and appear to involve both G protein- and AC-related changes. Endurance training was able to significantly offset these declines through restoration of the G(i)/G(s) ratio and AC activity. PMID- 11500308 TI - Kinetic modeling of [(18)F]FDG in skeletal muscle by PET: a four-compartment five rate-constant model. AB - Various modeling strategies have been developed to convert regional [(18)F]fluorodeoxyglucose ([(18)F]FDG) concentration measured by positron emission tomography (PET) to a measurement of physiological parameters. However, all the proposed models have been developed and tested mostly for brain studies. The purpose of the present study is to select the most accurate model for describing [(18)F]FDG kinetics in human skeletal muscle. The database consists of basal and hyperinsulinemic-euglycemic studies performed in normal subjects. PET data were first analyzed by an input-output modeling technique (often called spectral analysis). These results provided guidelines for developing a compartmental model. A new model with four compartments and five rate constants (5K model) emerged as the best. By accounting for plasma and extracellular and intracellular kinetics, this model allows, for the first time, PET assessment of the individual steps of [(18)F]FDG kinetics in human skeletal muscle, from plasma to extracellular space to transmembrane transport into the cell to intracellular phosphorylation. Insulin is shown to affect transport and phosphorylation but not extracellular kinetics, with the transport step becoming the main site of control. The 5K model also allows definition of the domain of validity of the classic three-compartment three- or four-rate-constant models. These models are candidates for an investigative tool to quantitatively assess insulin control on individual metabolic steps in human muscle in normal and physiopathological states. PMID- 11500309 TI - High-altitude acclimation increases the triacylglycerol/fatty acid cycle at rest and during exercise. AB - High-altitude acclimation alters lipid metabolism during exercise, but it is unknown whether this involves changes in rates of lipolysis or reesterification, which form the triacylglycerol/fatty acid (TAG/FA) cycle. We combined indirect calorimetry with [2-(3)H]glycerol and [1-(14)C]palmitate infusions to simultaneously measure total lipid oxidation, lipolysis, and rate of appearance (R(a)) of nonesterified fatty acids (NEFA) in high-altitude-acclimated (HA) rats exercising at 60% maximal O(2) uptake (VO(2 max)). During exercise, relative total lipid oxidation (%VO(2)) equaled sea-level control (SL) values; however, acclimation greatly stimulated lipolysis (+75%) but had no effect on R(a) NEFA. As a result, TAG/FA cycling increased (+119%), due solely to an increase in recycling (+144%) within adipocytes. There was no change in either group in these variables with the transition from rest to exercise. We conclude that, in HA, 1) acclimation is a potent stimulator of lipolysis; 2) rats do not modify TAG/FA cycling with the transition to exercise; and 3) in normoxia, HA and SL derive the same fraction of their total energy from lipids and carbohydrates. PMID- 11500310 TI - Parathyroid hormone receptor internalization is independent of protein kinase A and phospholipase C activation. AB - Parathyroid hormone (PTH) and PTH-related peptide (PTHrP) binding to their common receptor stimulates second messenger accumulation, receptor phosphorylation, and internalization. LLC-PK(1) cells expressing a green fluorescent protein-tagged PTH/PTHrP receptor show time- and dose-dependent receptor internalization. The internalized receptors colocalize with clathrin-coated pits. Internalization is stimulated by PTH analogs that bind to and activate the PTH/PTHrP receptor. Cell lines expressing a mutant protein kinase A regulatory subunit that is resistant to cAMP and/or a mutant receptor (DSEL mutant) that does not activate phospholipase C internalize their receptors normally. In addition, internalization of the wild-type receptor and the DSEL mutant is stimulated by the PTH analog [Gly(1),Arg(19)]hPTH-(1-28), which does not stimulate phospholipase C. Forskolin, IBMX, and the active phorbol ester, phorbol-12 myristate-13-acetate, did not promote receptor internalization or increase PTH induced internalization. These data indicate that ligand-induced internalization of the PTH/PTHrP receptor requires both ligand binding and receptor activation but does not involve stimulation of adenylate cyclase/protein kinase A or phospholipase C/protein kinase C. PMID- 11500311 TI - Effects of vitamin D receptor inactivation on the expression of calbindins and calcium metabolism. AB - Hypocalcemia, rickets, and osteomalacia are major phenotypic abnormalities in vitamin D receptor (VDR)-null mice. In an attempt to understand the abnormal regulation of calcium metabolism in these animals, we examined the expression of calbindins (CaBP) as well as calcium handling in the intestine and kidney of VDR null mice. In adult VDR-null mice, intestinal and renal CaBP-D9k expression was reduced by 50 and 90%, respectively, at both the mRNA and protein levels compared with wild-type littermates, whereas renal CaBP-D28k expression was not significantly changed. Intestinal calcium absorption was measured by the rate of (45)Ca disappearance from the intestine after an oral dose of the isotope. (45)Ca absorption was similar in VDR-null and wild-type mice, but the amount of (45)Ca accumulated in the serum and bone was 3-4 times higher in wild-type mice than in VDR-null mice. Despite the hypocalcemia, the urinary excretion of calcium in VDR null mice was not different from that in wild-type mice. Moreover, 1 wk of a high calcium diet treatment that normalized the serum ionized calcium level of VDR null mice increased the urinary calcium level of these mutant mice to twofold higher than that of wild-type mice on the same diet, suggesting impaired renal calcium conservation in VDR-null mice. These data demonstrate that renal CaBP D9k, but not CaBP-D28k, is highly regulated by the VDR-mediated action of 1,25 dihydroxyvitamin D(3). Furthermore, the results also suggest that impaired calcium conservation in the kidney may be the most important factor contributing to the development of hypocalcemia in VDR-null mice, and CaBP-D9k may be an important mediator of calcium reabsorption in the kidney. PMID- 11500312 TI - Amino acids and insulin are both required to regulate assembly of the eIF4E. eIF4G complex in rat skeletal muscle. AB - The respective roles of insulin and amino acids in regulation of skeletal muscle protein synthesis and degradation after feeding were examined in rats fasted for 17 h and refed over 1 h with either a 25 or a 0% amino acid/protein meal. In each nutritional condition, postprandial insulin secretion was either maintained (control groups: C(25) and C(0)) or blocked with diazoxide injections (diazoxide groups: DZ(25) and DZ(0)). Muscle protein metabolism was examined in vitro in epitrochlearis muscles. Only feeding the 25% amino acid/protein meal in the presence of increased plasma insulin concentration (C(25) group) stimulated protein synthesis and inhibited proteolysis in skeletal muscle compared with the postabsorptive state. The stimulation of protein synthesis was associated with increased phosphorylation of eukaryotic initiation factor (eIF)4E binding protein 1 (4E-BP1), reduced binding of eIF4E to 4E-BP1, and increased assembly of the active eIF4E. eIF4G complex. The p70 S6 kinase (p70(S6k)) was also hyperphosphorylated in response to the 25% amino acid/protein meal. Acute postprandial insulin deficiency induced by diazoxide injections totally abolished these effects. Feeding the 0% amino acid/protein meal with or without postprandial insulin deficiency did not stimulate muscle protein synthesis, reduce proteolysis, or regulate initiation factors and p70(S6k) compared with fasted rats. Taken together, our results suggest that both insulin and amino acids are required to stimulate protein synthesis, inhibit protein degradation, and regulate the interactions between eIF4E and 4E-BP1 or eIF4G in response to feeding. PMID- 11500313 TI - Type 1 diabetes leads to cytoskeleton changes that are reflected in insulin action on rat cardiac K(+) currents. AB - A sustained K(+) current (I(ss)) is attenuated in ventricular cells from streptozotocin (STZ)-induced diabetic rats. The in vitro addition of insulin to isolated cells augments I(ss) in a process that is blocked by disrupting either actin microfilaments (with cytochalasin D) or microtubules (with colchicine). When these agents are added at progressively later times, the effect of insulin becomes evident in a time-dependent manner. I(ss) is also augmented by insulin in control cells in a cytoskeleton-dependent manner. However, in contrast to diabetic cells, cytoskeleton-dependent augmentation of I(ss) by insulin occurs at a considerably faster rate in control cells. Immunofluorescent labeling shows a reduced density of beta-tubulin in diabetic cells, particularly in perinuclear regions. In vitro insulin replacement or in vivo insulin injections given to STZ treated rats enhances beta-tubulin density. These results suggest an impairment of cytoskeleton function and structure under insulin-deficient conditions, which may have implications for cardiac function. PMID- 11500314 TI - Body surface area prediction in normal-weight and obese patients. AB - None of the equations frequently used to predict body surface area (BSA) has been validated for obese patients. We applied the principles of body size scaling to derive an improved equation predicting BSA solely from a patient's weight. Forty five patients weighing from 51.3 to 248.6 kg had their height and weight measured on a calibrated scale and their BSA calculated by a geometric method. Data were combined with a large series of published BSA estimates. BSA prediction with the commonly used Du Bois equation underestimated BSA in obese patients by as much as 20%. The equation we derived to relate BSA to body weight was a power function: BSA (m(2)) = 0.1173 x Wt (kg)(0.6466). Below 10 kg, this equation deviated significantly from the BSA vs. body weight curve, necessitating a different set of coefficients: BSA (m(2)) = 0.1037 x Wt (kg)(0.6724). Covariance of height and weight for patients weighing <80 kg reduced the Du Bois BSA-predicting equation to a power function, explaining why it provides good BSA predictions for normal size patients but fails with obesity. PMID- 11500315 TI - Glucocorticoids impair fetal beta-cell development in rats. AB - In rats, poor fetal growth due to maternal food restriction during pregnancy is associated with decreased beta-cell mass at birth and glucose intolerance in adulthood. Overexposure to glucocorticoids in utero can induce intrauterine growth retardation in humans and animals and subsequent glucose intolerance in rodents. The aims of this study were to investigate whether glucocorticoid overexposure mediates the effect of undernutrition on beta-cell mass and to study their potential role in normally nourished rats. Undernutrition significantly increased maternal and fetal corticosterone levels. Twenty-one-day-old fetuses with undernutrition showed growth retardation and decreased pancreatic insulin content; adrenalectomy and subcutaneous corticosterone implants in their dams prevented the maternal corticosterone increase and restored fetal beta-cell mass. In fetuses with normal nutrition, fetal corticosterone levels were negatively correlated to fetal weight and insulin content; fetal beta-cell mass increased from 355 +/- 48 microg in sham to 516 +/- 160 microg after maternal adrenalectomy; inhibition of steroid production by metyrapone induced a further increase to 757 +/- 125 microg. Our data support the new concept of a negative role of glucocorticoids in fetal beta-cell development. PMID- 11500316 TI - Insulin increases FA uptake and esterification but reduces lipid utilization in isolated contracting muscle. AB - We examined the effect of insulin on the synthesis and degradation of muscle lipid pools [phospholipid (PL), diacylglycerol (DG), triacylglycerol (TG)] and palmitate oxidation in isolated resting and contracting (20 tetani/min) soleus muscles. Lipid metabolism was monitored using the previously defined pulse-chase procedure. At rest, insulin significantly increased total palmitate uptake into soleus muscle (+49%, P < 0.05), corresponding to enhanced DG (+60%, P < 0.05) and TG (+61%, P < 0.05) esterification, but blunted palmitate oxidation (-38%, P < 0.05) and TG hydrolysis (-34%, P < 0.05). During muscle contraction, when total palmitate uptake was increased, insulin further enhanced uptake (+21%, P < 0.05) and esterification of fatty acids (FA) to PL (+73%, P < 0.05), DG (+19%, P < 0.05), and TG (+161%, P < 0.01). Despite a profound shift in the relative partitioning of FA away from esterification and toward oxidation during contraction, the increase in palmitate oxidation and TG hydrolysis was significantly blunted by insulin [oxidation, -24% (P = 0.05); hydrolysis, -83% (P < 0.01)]. The effects of insulin on FA esterification (stimulation) and oxidation (inhibition) during contraction were reduced in the presence of the phosphatidylinositol 3-kinase inhibitor LY-294002. In summary, the effects of insulin and contraction on palmitate uptake and esterification are additive, while insulin opposes the stimulatory effect of contraction on FA oxidation and TG hydrolysis. Insulin's modulatory effects on muscle FA metabolism during contraction are mediated at least in part through phosphatidylinositol 3-kinase. PMID- 11500317 TI - GLUT-4 translocation in skeletal muscle studied with a cell-free assay: involvement of phospholipase D. AB - GLUT-4-containing membranes immunoprecipitated from insulin-stimulated rat skeletal muscle produce the phospholipase D (PLD) product phosphatidic acid. In vitro stimulation of PLD in crude membrane with ammonium sulfate (5 mM) resulted in transfer of GLUT-4 (3.0-fold vs. control) as well as transferrin receptor proteins from large to small membrane structures. The in vitro GLUT-4 transfer could be blocked by neomycin (a PLD inhibitor), and neomycin also reduced insulin stimulated glucose transport in intact incubated soleus muscles. Furthermore, protein kinase B(beta) (PKB(beta)) was found to associate with the GLUT-4 protein and was transferred to small vesicles in response to ammonium sulfate in vitro. Finally, addition of cytosolic proteins, prepared from basal skeletal muscle, and GTP nucleotides to an enriched GLUT-4 membrane fraction resulted in in vitro transfer of GLUT-4 to small membranes (6.8-fold vs. unstimulated control). The cytosol and nucleotide-induced GLUT-4 transfer could be blocked by neomycin and N ethylmaleimide. In conclusion, we have developed a cell-free assay that demonstrates in vitro GLUT-4 transfer. This transfer may suggest release of GLUT 4-containing vesicles from donor GLUT-4 membranes involving PLD activity and binding of PKB(beta) to GLUT-4. PMID- 11500318 TI - Impaired muscular contractile performance and adenine nucleotide handling in creatine kinase-deficient mice. AB - Creatine kinase (CK) forms a small family of isoenzymes playing an important role in maintaining the concentration of ATP and ADP in muscle cells. To delineate the impact of a lack of CK activity, we studied contractile performance during a single maximal tetanic contraction and during 12 repeated tetanic contractions of intact dorsal flexors of CK knockout (CK(-/-)) mice. To investigate the effect on ATP regeneration, muscular high-energy phosphate content was determined at rest, immediately after the contraction series, and after a 60-s recovery period. Maximal torque of the dorsal flexors was significantly lower in CK(-/-) mice than in wild-type animals, i.e., 23.7 +/- 5.1 and 33.3 +/- 6.8 mN. m. g(-1) wet wt, respectively. Lower muscle ATP (20.1 +/- 1.4 in CK(-/-) vs. 28.0 +/- 2.1 micromol/g dry wt in controls) and higher IMP (1.2 +/- 0.5 in CK(-/-) vs. 0.3 +/- 0.1 micromol/g dry wt in controls) levels at the onset of contraction may contribute to the declined contractility in CK(-/-) mice. In contrast to wild type muscles, ATP levels could not be maintained during the series of 12 tetanic contractions of dorsal flexors of CK(-/-) mice and dropped to 15.5 +/- 2.4 micromol/g dry wt. The significant increase in tissue IMP (2.4 +/- 1.1 micromol/g dry wt) content after the contraction series indicates that ATP regeneration through adenylate kinase was not capable of fully compensating for the lack of CK. ATP regeneration via the adenylate kinase pathway is a likely cause of reduced basal adenine nucleotide levels in CK(-/-) mice. PMID- 11500320 TI - Age-related decline in RMR in physically active men: relation to exercise volume and energy intake. AB - We tested the hypothesis that resting metabolic rate (RMR) declines with age in physically active men (endurance exercise > or =3 times/wk) and that this decline is related to weekly exercise volume (h/wk) and/or daily energy intake. Accordingly, we studied 137 healthy adult men who had been weight stable for > or =6 mo: 32 young [26 +/- 1 (SE) yr] and 34 older (62 +/- 1 yr) sedentary males (internal controls); and 39 young (27 +/- 1 yr) and 32 older (63 +/- 2 yr) physically active males (regular endurance exercise). RMR was measured by indirect calorimetry (ventilated hood system) after an overnight fast and approximately 24 h after exercise. Because RMR is related to fat-free mass (FFM; r = 0.76, P < 0.001, current study), FFM was covaried to adjust RMR (RMR(adj)). RMR(adj) was lower with age in both the sedentary (72.0 +/- 2.0 vs. 64.0 +/- 1.3 kcal/h, P < 0.01) and the physically active (76.6 +/- 1.1 vs. 67.9 +/- 1.2 kcal/h, P < 0.01) males. In the physically active men, RMR(adj) was related to both exercise volume (no. of h/wk, regardless of intensity; r = 0.56, P < 0.001) and estimated energy intake (r = 0.58, P < 0.001). Consistent with these relations, RMR(adj) was not significantly different in subgroups of young and older physically active men matched either for exercise volume (h/wk; n = 11 each) or estimated energy intake (kcal/day; n = 6 each). These results indicate that 1) RMR, per unit FFM, declines with age in highly physically active men; and 2) this decline is related to age-associated reductions in exercise volume and energy intake and does not occur in men who maintain exercise volume and/or energy intake at a level similar to that of young physically active men. PMID- 11500319 TI - Postprandial dyslipidemia in men with visceral obesity: an effect of reduced LDL receptor expression? AB - Postprandial lipemia after an oral fat challenge was studied in middle-aged men with visceral obesity. The two groups had similar plasma cholesterol levels, but obese subjects had higher levels of plasma triglyceride and reduced amounts of high-density cholesterol. Fasting plasma insulin was fourfold greater in obese subjects because of concomitant insulin resistance, with a calculated HOMA score of 3.1 +/- 0.6 vs. 0.8 +/- 0.2, respectively. Plasma apolipoprotein B(48) (apoB(48)) and retinyl palmitate (RP) after an oral fat challenge were used to monitor chylomicron metabolism. Compared with lean subjects, the fasting concentration of apoB(48) was more than twofold greater in obese individuals, suggestive of an accumulation of posthydrolyzed particles. After the oral lipid load, the incremental areas under the apoB(48) and RP curves (IAUC) were both significantly greater in obese subjects (apoB(48): 97 +/- 17 vs. 44 +/- 12 microg.ml(-1). h; RP: 3,120 +/- 511 vs. 1,308 +/- 177 U. ml(-1). h, respectively). A delay in the conversion of chylomicrons to remnants probably contributed to postprandial dyslipidemia in viscerally obese subjects. The triglyceride IAUC was 68% greater in obese subjects (4.7 +/- 0.6 vs. 2.8 +/- 0.8 mM. h, P < 0.06). Moreover, peak postprandial triglyceride was delayed by approximately 2 h in obese subjects. The reduction in triglyceride lipolysis in vivo did not appear to reflect changes in hydrolytic enzyme activities. Postheparin plasma lipase rates were found to be similar for lean and obese subjects. In this study, low-density lipoprotein (LDL) receptor expression on monunuclear cells was used as a surrogate marker of hepatic activity. We found that, in obese subjects, the binding of LDL was reduced by one-half compared with lean controls (70.9 +/- 15.07 vs. 38.9 +/- 4.6 ng LDL bound/microg cell protein, P = 0.02). Because the LDL receptor is involved in the removal of proatherogenic chylomicron remnants, we suggest that the hepatic clearance of these particles might be compromised in insulin-resistant obese subjects. Premature and accelerated atherogenesis in viscerally obese, insulin-resistant subjects may in part reflect delayed clearance of postprandial lipoprotein remnants. PMID- 11500321 TI - Programming into adulthood of islet adaptations induced by early nutritional intervention in the rat. AB - To investigate the influence of a high carbohydrate (HC) intake during the suckling period on pancreatic function in adult life, neonatal rats were artificially reared on a HC milk formula during the preweaning period and then weaned onto lab chow. In the adult HC rat, hyperinsulinemia is sustained by a variety of biochemical and molecular adaptations induced in the HC islets during the suckling period. The adult HC islets showed a distinct left shift in the glucose-stimulated insulin-secretory pattern. HC islets were also able to secrete moderate levels of insulin in the absence of glucose and in the presence of Ca(2+) channel inhibitors. In addition, the mRNA levels of preproinsulin, somatostatin transcription factor-1, upstream stimulatory factor-1, stress activated protein kinase-2, phosphatidylinositol kinase, and GLUT-2 genes were significantly increased in HC islets. These results show that consumption of a HC formula during the suckling period programs pancreatic islet function in adult rats, resulting in the maintenance of hyperinsulinemia in the postweaning period and eventually leading to the development of obesity in adult life. PMID- 11500328 TI - Sleep in space flight: breath easy--sleep less? PMID- 11500329 TI - Prevention of ventilator-associated pneumonia by oral decontamination: just another SDD study? PMID- 11500330 TI - End-stage chronic obstructive pulmonary disease: the cigarette is burned out but inflammation rages on. PMID- 11500331 TI - Critical care neurology. PMID- 11500332 TI - Sudden infant death syndrome and other causes of infant mortality: diagnosis, mechanisms, and risk for recurrence in siblings. PMID- 11500333 TI - Lower respiratory illnesses promote FEV(1) decline in current smokers but not ex smokers with mild chronic obstructive pulmonary disease: results from the lung health study. AB - We analyzed Lung Health Study (LHS) data to assess the effect of self-reported lower respiratory illnesses resulting in physician visits (LRI) on lung function. Participants were 5,887 smokers aged 35-60 yr, FEV(1)/FVC < 0.70 and FEV(1) of 55 90% predicted. Two-thirds were randomized into an intensive smoking cessation program (SI); one-third were advised only to stop smoking (UC). For 5 yr participants had annual spirometry and questioning regarding LRI. SI had greater rates of smoking cessation than usual care (UC) with fewer LRI (p = 0.0008). Sustained quitters had fewer LRI than continuing smokers (p = 0.0003). In the year LRI occurred, FEV(1) did not change in sustained quitters, but decreased significantly in smokers (p = 0.0001) with some recovery the following year if no LRI occurred. Over 5 yr, LRI had a significant effect on rate of decline of FEV(1) only in smokers. In smokers averaging one LRI/yr over 5 yr there were additional declines in FEV(1) of 7 ml /yr (p = 0.001). Smokers with more than one LRI/yr had greater declines. Chronic bronchitis was associated with increased frequencies of LRI, but did not affect their influence on lung function. Smoking and LRI had an interactive effect on FEV(1) in people with mild COPD, and in smokers frequent LRI may influence the long-term course of the disease. PMID- 11500334 TI - Effects of withdrawal of inhaled steroids in men with severe irreversible airflow obstruction. AB - Inhaled corticosteroid therapy has proven efficacy for asthmatics, but the benefit for patients with chronic obstructive pulmonary disease (COPD) is less well supported. We hypothesized that withdrawal of inhaled steroids in elderly patients with severe irreversible airway obstruction would not lead to a deterioration in respiratory function. We designed a prospective, double-blind, randomized, placebo-controlled, crossover study to follow spirometry, quality of life questionnaire, six-minute (6-min) walk test, and sputum markers of inflammation during a 6-wk placebo treatment period and a 6-wk treatment period with beclomethasone dipropionate (BDP), 336 microg/d. There were 24 men receiving BDP who entered the study; 15 completed the study. Their mean age was 66.9 +/- 1.9 yr, and mean FEV(1) was 1.61 +/- 0.1 L (47% of predicted). There was a significant decrease in the mean FEV(1 )while using the placebo inhaler (1.70 L versus 1.60 L, baseline versus placebo: 95% CI, 0.002 to 0.195; p < 0.05). There was a decrease in the mean percentage change in FEV(1) for the study subjects during the placebo treatment period as compared with the BDP treatment period ( 6.28 versus 5.03%, placebo versus BDP: 95% CI, -23.38 to 0.76; p = 0.06). Six minute walk test results and sputum analysis for cell count and differential were not significantly different during placebo and BDP treatment periods. Borg scale assessment of dyspnea after exercise was increased while using the placebo inhaler as compared with baseline, and decreased during the BDP treatment period. Chronic Respiratory Disease Questionnaire (CRQ) scores revealed no significant difference between placebo and BDP. This study has demonstrated that in elderly patients with severe irreversible airway obstruction, withdrawal of inhaled corticosteroid therapy leads to a deterioration in ventilatory function and increased exercise-induced dyspnea. PMID- 11500335 TI - The health impact of undiagnosed airflow obstruction in a national sample of United States adults. AB - To determine the health and functional impact of undiagnosed airflow obstruction for subjects in the general population, we used data obtained as part of the Third National Health and Nutrition Examination Survey (NHANES III). Categories of diagnosed and undiagnosed airflow obstruction were defined using questionnaire responses and spirometric results. Health and functional impact of airflow obstruction was assessed from responses to questions about general health status, walking 1/4 mile, lifting or carrying something as heavy as 10 lb, or needing help with personal care. Undiagnosed airflow obstruction (12.0%) was more common than doctor-diagnosed chronic obstructive pulmonary disease (COPD) (3.1%) or asthma (2.7%). Although undiagnosed airflow obstruction was usually very mild, approximately 5% of the entire sample had an FEV(1) less than 75% predicted. After adjusting for smoking, obesity, and comorbid conditions, the risk of impaired health and functional status with undiagnosed airflow obstruction was independently associated with severity of FEV(1) impairment. For males and females, ever smoking was strongly associated with all types of airflow obstruction, diagnosed or not. However, among females with airflow obstruction, 12.2% to 35.2% never smoked. Undiagnosed airflow obstruction is common in the general population of the United States and is associated with impaired health and functional status. KEYWORDS: airflow obstruction; spirometry; health impact; screening PMID- 11500336 TI - Validity and safety of sputum induction by inhaled uridine 5'-triphosphate. AB - Inhalation of hypertonic saline during sputum induction causes bronchoconstriction. We studied the validity and safety of sputum induction by uridine 5'-triphosphate (UTP). Sputum was induced by a 5-min inhalation of hypertonic saline (3%) on Day 1 and UTP (5 mg/ml in 0.9% saline) on Days 8 and 15 in 16 healthy subjects and 16 patients with mild-to-moderate asthma. Inhaled UTP produced twofold greater amounts of sputum than did hypertonic saline. There were significant differences in oxygen desaturation and bronchoconstriction during the procedure between the two methods: the maximal fall in Sa(O(2)), the AUC of the Sa(O(2))-time response, and the fall in PEF were less in the subjects who received UTP than in those who received hypertonic saline. Sputum total cell and differential cell counts, with a high proportion of eosinophils in asthmatics, were similar between specimens obtained by hypertonic saline and UTP. When we compared two consecutive measurements on the UTP-induced sputum samples, the reproducibility calculated by the intraclass correlation coefficient was high for the proportion of eosinophils, neutrophils, and macrophages. Therefore, inhalation of UTP aerosols may provide an effective, relatively noninvasive, valid, and reproducible method of sputum induction for the assessment of airway inflammation in asthma. KEYWORDS: uridine triphosphate; induced sputum; airway inflammation; bronchoconstriction; asthma PMID- 11500337 TI - Prevention of ventilator-associated pneumonia by oral decontamination: a prospective, randomized, double-blind, placebo-controlled study. AB - Colonization of the intestinal tract has been assumed to be important in the pathogenesis of ventilator-associated pneumonia (VAP), but relative impacts of oropharyngeal, gastric, or intestinal colonization have not been elucidated. Our aim was to prevent VAP by modulation of oropharyngeal colonization, without influencing gastric and intestinal colonization and without systemic prophylaxis. In a prospective, randomized, placebo-controlled, double-blind study, 87 patients received topical antimicrobial prophylaxis (gentamicin/ colistin/vancomycin 2% in Orabase, every 6 h) in the oropharynx and 139 patients, divided over two control groups, received placebo (78 patients were studied in the presence of patients receiving topical prophylaxis [control group A] and 61 patients were studied in an intensive care unit where no topical prophylaxis was used [control group B]). Baseline characteristics were comparable in all three groups. Topical prophylaxis eradicated colonization present on admission in oropharynx (75% in study group versus 0% in control group A [p < 0.00001] and 9% in control group B patients [p < 0.00001]) and in trachea (52% versus 22% in A [p = 0.03] and 7% in B [p = 0.004]). Moreover, topical prophylaxis prevented acquired oropharyngeal colonization (10% versus 59% in A [p < 0.00001] and 63% in B [p < 0.00001]). Colonization rates in stomach and intestine were not affected. Incidences of VAP were 10% in study patients, 31% in Group A, and 23% in Group B patients (p = 0.001 and p = 0.04, respectively). This was not associated with shorter durations of ventilation or ICU stay or better survival. Oropharyngeal colonization is of paramount importance in the pathogenesis of VAP, and a targeted approach to prevent colonization at this site is a very effective method of infection prevention. KEYWORDS: cross infection, prevention and control; respiration, artificial, adverse effects; antibiotics, administration and dosage infection control methods; pneumonia, etiology, prevention and control; intubation, intratracheal, adverse effects PMID- 11500338 TI - Mitochondrial membrane potential and apoptosis peripheral blood monocytes in severe human sepsis. AB - Reduced mitochondrial membrane potential (Delta(Psi)m), which is considered as an initial and irreversible step towards apoptosis, as well as cell death regulating proteins, such as Fas, Hsp70, or Bcl-2, may play an important role in sepsis. We studied the relationship between sepsis severity and peripheral blood monocyte Delta(Psi)m, cell death (necrosis and apoptosis), soluble Fas ligand, Hsp70, and Bcl-2 expression over time in 18 patients with sepsis, and compared these data with those of a group of 17 healthy control subjects. All measurements were performed within 3 d of the onset of severe sepsis (T1), then 7 to 10 d later (T2), and finally at hospital discharge (T3). Delta(Psi)m was expressed as the percent monocytes with altered Delta(Psi)m (%Delta(Psi)m). Patients with sepsis had greater %Delta(Psi)m at T1 and T2 but not at T3 (14.6 +/- 2.6% and 15.9 +/- 2%, respectively, versus control 6.6 +/- 0.2%, p < 0.01). Septic patients exhibited greater cell death in their monocytes and had greater Hsp70 expression only at T1. Bcl-2 levels were similar in septic and control subjects. Comparing survivors with non-survivors of sepsis, nonsurvivors had a greater %Delta(Psi)m at T1 (26.4 +/- 5.3% versus 10.1 +/- 2.7%, p < 0.01) and a significant decrease in Bcl-2 expression, whereas no difference was found in Hsp70 levels. These results indicate that mitochondrial dysfunction and subsequent cell death occur in severe sepsis and suggest that %Delta(Psi)m is a marker of severity in human sepsis. KEYWORDS: mitochondria; apoptosis; sepsis; heat-shock protein 70; proto oncogene protein c-Bcl-2 PMID- 11500339 TI - Diagnostic value of procalcitonin, interleukin-6, and interleukin-8 in critically ill patients admitted with suspected sepsis. AB - To assess the diagnostic value of procalcitonin (PCT), interleukin (IL)-6, IL-8, and standard measurements in identifying critically ill patients with sepsis, we performed prospective measurements in 78 consecutive patients admitted with acute systemic inflammatory response syndrome (SIRS) and suspected infection. We estimated the relevance of the different parameters by using multivariable regression modeling, likelihood-ratio tests, and area under the receiver operating characteristic curves (AUC). The final diagnosis was SIRS in 18 patients, sepsis in 14, severe sepsis in 21, and septic shock in 25. PCT yielded the highest discriminative value, with an AUC of 0.92 (CI, 0.85 to 1.0), followed by IL-6 (0.75; CI, 0.63 to 0.87), and IL-8 (0.71; CI, 0.59 to 0.83; p < 0.001). At a cutoff of 1.1 ng/ml, PCT yielded a sensitivity of 97% and a specificity of 78% to differentiate patients with SIRS from those with sepsis-related conditions. Median PCT concentrations on admission (ng/ ml, range) were 0.6 (0 to 5.3) for SIRS; 3.5 (0.4 to 6.7) for sepsis; 6.2 (2.2 to 85) for severe sepsis; and 21.3 (1.2 to 654) for septic shock (p < 0.001). The addition of PCT to a model based solely on standard indicators improved the predictive power of detecting sepsis (likelihood ratio test; p = 0.001) and increased the AUC value for the routine value-based model from 0.77 (CI, 0.64 to 0.89) to 0.94 (CI, 0.89 to 0.99; p = 0.002). In contrast, no additive effect was seen for IL-6 (p = 0.56) or IL-8 (p = 0.14). Elevated PCT concentrations appear to be a promising indicator of sepsis in newly admitted, critically ill patients capable of complementing clinical signs and routine laboratory parameters suggestive of severe infection. PMID- 11500340 TI - Ultrasonic examination: an alternative to chest radiography after central venous catheter insertion? AB - We evaluated ultrasonic examination as a diagnostic tool for catheter misplacement and pneumothorax after central venous catheter insertion. Physicians in the intensive care unit (ICU) performed the ultrasonic examinations, and the results were compared with those of chest radiography. Eighty-five central venous catheters (70 subclavian and 15 internal jugular) were inserted into 81 patients; 10 misplacements and one pneumothorax occurred. Ultrasonic examination feasibility was 99.6%. The only pneumothorax and all misplacements except one were diagnosed by ultrasound. Taking into consideration misplacements and pneumothorax research, ultrasonic examination did not give any false positive results. The mean time of the entire ultrasonic examination was 6.8 +/- 3.5 min, whereas 80.3 +/- 66.7 min were needed for the radiography (p < 0.0001). This study has suggested that ultrasonic diagnosis of catheter misplacement and pneumothorax related to central venous catheterization is a rapid and accurate method that can be easily performed by ICU physicians. PMID- 11500341 TI - Alveolar JE/MCP-1 and endotoxin synergize to provoke lung cytokine upregulation, sequential neutrophil and monocyte influx, and vascular leakage in mice. AB - The C-C chemokine monocyte chemotactic protein 1 (JE/MCP-1) is a key cytokine for lung monocyte recruitment, and may be detected in high levels in the alveolar space in lung injury. We hypothesized that alveolar JE/MCP-1 might synergize with endotoxin in this compartment to elicit lung inflammatory events. Intratracheal instillation of JE/MCP-1 into BALB/c mice did not provoke increased bronchoalveolar lavage tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL 6), and macrophage inflammatory protein 2 (MIP-2) levels, but elicited monocyte recruitment into this compartment. Intratracheal Escherichia coli endotoxin provoked elevated lavage TNF-alpha, IL-6, and MIP-2 levels, peaking after 6 h in parallel with increased alveolar neutrophil numbers, in the absence of vascular leakage. Mice receiving both endotoxin and JE/ MCP-1 showed drastically increased lavage TNF-alpha, IL-6, and MIP-2 levels, 22-fold higher lavage neutrophil numbers, and lung vascular leakage. Moreover, an 8-fold increased alveolar accumulation of monocytes, peaking at 48 h together with expansion of the resident alveolar macrophage pool, was noted. Intraperitoneal instead of alveolar deposition of MCP-1 or endotoxin failed to reproduce the synergistic response, and the same was true for employment of RANTES instead of MCP-1. Blockade of neutrophil recruitment by anti-CD18 did not affect the intra-alveolar cytokine response to MCP-1 plus endotoxin. Together, JE/MCP-1 and endotoxin, when coappearing in the alveolar compartment at low dosage, elicit an early phase of lung inflammatory injury with increased cytokine synthesis and neutrophil recruitment, and a late phase of enhanced monocyte traffic and expansion of the alveolar macrophage pool. PMID- 11500342 TI - Endogenous nitric oxide synthesis and vascular leakage in ischemic-reperfused rabbit lungs. AB - Pulmonary edema formation resulting from loss of capillary barrier properties is a prominent finding in lung ischemia/reperfusion (I/R) injury. The role of endogenous nitric oxide (NO) in this process is unresolved. We exposed buffer perfused rabbit lungs to warm I/R and measured air space NO liberation and intravascular accumulation of NO degradation products. In lungs undergoing 210 min of ischemia with normoxic ventilation, with maintenance of positive intravascular pressure to avoid vascular collapse, NO synthesis was moderately reduced during ischemia but was fully restored upon reperfusion, and a moderate leakage response occurred during reperfusion. Pretreatment with the NO synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA) suppressed NO synthesis but did not affect the leakage. During ischemia with anoxic ventilation, NO synthesis was fully abrogated, but again promptly reappeared upon reperfusion and entrance of oxygen into the system. It was with this protocol that the most severe vascular leakage was encountered, which was markedly reduced in the presence of L-NMMA or superoxide dismutase. We conclude that endogenous NO does not play a major role in the induction or mitigation of I/R injury under conditions of normoxic ischemia, but that return of endogenous NO synthesis upon reperfusion after anoxic ischemia contributes substantially to the triggering of vascular leakage, possibly via interaction with superoxide. PMID- 11500343 TI - Electrical activity of the diaphragm during pressure support ventilation in acute respiratory failure. AB - We compared crural diaphragm electrical activity (EAdi) with transdiaphragmatic pressure (Pdi) during varying levels of pressure support ventilation (PS) in 13 intubated patients. With changing PS, we found no evidence for changes in neuromechanical coupling of the diaphragm. From lowest to highest PS (2 cm H(2)O +/- 4 to 20 cm H(2)O +/- 7), tidal volume increased from 430 ml +/- 180 to 527 ml +/- 180 (p < 0.001). The inspiratory volume calculated during the period when EAdi increased to its peak did not change from 276 +/- 147 to 277 +/- 162 ml, p = 0.976. Respiratory rate decreased from 23.9 (+/- 7) to 21.3 (+/- 7) breaths/min (p = 0.015). EAdi and Pdi decreased proportionally by adding PS (r = 0.84 and r = 0.90, for mean and peak values, respectively). Mean and peak EAdi decreased (p < 0.001) by 33 +/- 21% (mean +/- SD) and 37 +/- 23% with the addition of 10 cm H(2)O of PS, similar to the decrease in the mean and peak Pdi (p < 0.001) observed (34 +/- 36 and 35 +/- 23%). We also found that ventilator assist continued during the diaphragm deactivation period, a phenomenon that was further exaggerated at higher PS levels. We conclude that EAdi is a valid measurement of neural drive to the diaphragm in acute respiratory failure. PMID- 11500344 TI - Unexplained exertional limitation: characterization of patients with a mitochondrial myopathy. AB - Exercise intolerance is a common complaint, the cause of which often remains elusive after a comprehensive evaluation. In this report, we describe 28 patients with unexplained dyspnea or exertional limitation secondary to biopsy-proven mitochondrial myopathies. Patients were prospectively identified from a multidisciplinary dyspnea clinic at a tertiary referral center. All patients were without underlying pulmonary, cardiac, or other neuromuscular disorders. Patients underwent history, physical examination, complete pulmonary function testing, respiratory muscle testing, cardiopulmonary exercise testing, and muscle biopsy. Results were compared with a group of normal control subjects. The estimated period prevalence was 8.5% (28 of 331). Spirometry, lung volumes, and gas exchange were normal in patients and control subjects. Compared with control subjects, the patient group demonstrated decreased exercise capacity (maximum achieved V O(2) 67 versus 104% predicted; p < 0.0001) and respiratory muscle weakness (PI(max) 77 versus 115% predicted; p = 0.001). These patients have a characteristic exercise response that was hyperventilatory (peak VE/V CO(2); 55 versus 42) and hypercirculatory (maximum heart rate - baseline heart rate/V O(2)max - baseline V O(2)max; 91 versus 41) compared to control subjects. Patients stopping exercise due to dyspnea (n = 16) (as compared with muscle fatigue, n = 11) displayed weaker respiratory muscles (Pdi(max) 61 versus 115 cm H(2)O; p = 0.01) and were more likely to reach mechanical ventilatory limitation (V Emax/ MVV 0.81 versus 0.58; p = 0.02). The sensation of dyspnea was related to indices of respiratory muscle function including respiratory rate and inspiratory flow. We conclude that mitochondrial myopathies are more prevalent than previously reported. The characteristic physiological profile may be useful in the diagnostic evaluation of mitochondrial myopathy. PMID- 11500345 TI - Effect of maturity on maximal transdiaphragmatic pressure in infants during crying. AB - The aim of this study was to determine the effect of maturation on diaphragmatic function. In addition, we investigated whether noninvasive assessment yielded similar results to invasive measurement. Twenty-eight infants, median gestational age (GA) 35.5 wk (range, 25 to 42 wk) and postconceptional age (PCA), 37.6 wk (range, 32 to 44 wk), were examined. Diaphragmatic function was assessed by measuring the maximal transdiaphragmatic pressure during crying (cPdi) using balloon catheters in the midesophagus (Pes) and the stomach (Pgas). In 14 of the infants, a noninvasive measurement of inspiratory muscle strength, maximal inspiratory pressure (PImax), was also made. cPdi and PImax were recorded during a crying effort with the airway occluded at end-expiration. The median cPdi and Pes during crying (cPes), but not Pgas during crying (cPgas), were significantly lower in those studied at a PCA of less than term compared with those studied at an older age (p < 0.05). cPdi and cPes, but not cPgas, correlated significantly with PCA (r = 0.44, p < 0.02; r = 0.43, p < 0.03; respectively) and gestational age (r = 0.46, p < 0.02 and r = 0.56, p < 0.01; respectively). In the 14 infants, the median PImax was lower, but it correlated significantly with cPdi (r = 0.79, p < 0.01). We conclude maturation does affect diaphragm function, and PImax may provide a noninvasive index of diaphragm strength. PMID- 11500346 TI - Role of nitric oxide in thermoregulation and hypoxic ventilatory response in obese Zucker rats. AB - To examine the role of nitric oxide (NO) on thermoregulation and control of breathing in obesity, awake obese and age-matched lean Zucker (Z) rats underwent a sustained hypoxic challenge. Body temperature (Tb), oxygen consumption (V O(2)) and ventilation (V E) were measured during room air and during 30-min of hypoxia (10% O(2)) after intraperitoneal administration of either 100 mg/kg of N(G)-nitro L-arginine methyl ester (L-NAME), a nonspecific NOS inhibitor, 25 mg/kg of 7 nitroindazole (7-NI), a selective neuronal NOS inhibitor, or equal volume of vehicle (dimethyl sulfoxide: DMSO) as control. Tb in obese rats during room air was significantly lower than that of lean rats. Hypoxia induced a more pronounced drop in Tb and V O(2) in lean rats than in obese rats. Tb in lean Z rats dropped significantly by approximately 0.2 degrees C after L-NAME and, more markedly, by approximately 1.1 degrees C after 7-NI compared with control during room air, whereas Tb in obese Z rats was unaffected. L-NAME and 7-NI attenuated hypoxia induced hypothermia or hypometabolism in lean rats, but not in obese rats. Lean rats exhibited an abrupt increase in V E in response to hypoxia followed by a gradual decline in V E. In contrast, obese rats displayed an initial increase in V E that plateaued during sustained hypoxia. Both L-NAME and 7-NI induced marked decreases in V E during room air and hypoxia compared with control lean rats, whereas V E was virtually unaffected by either agent in obese rats. The present results suggest that the blunted thermoregulatory and ventilatory responses to hypoxia in obese Z rats may be attributed to reduced activity of NOS in the central nervous system. PMID- 11500347 TI - Exercise inhibits epithelial sodium channels in patients with cystic fibrosis. AB - The aim of this study was to determine the effects of a single exercise bout on luminal Cl(-) and Na(+) conductance in the respiratory epithelium of patients with cystic fibrosis (CF). In nine patients with CF and nine healthy control subjects, the transepithelial electrical potential difference (PD) of the nasal respiratory epithelium was recorded, first at rest and then during moderate intensity exercise. Under both conditions, PD was first measured while superfusing the epithelium with isotonic saline. Then, the effects of amiloride and amiloride plus low chloride plus isoproterenol were determined. Exercise resulted in a significant lower PD compared with rest in patients with CF (-6.6 +/- 16.6 mV versus -33.6 +/- 10.0 mV, p < 0.0001) and control subjects (0.1 +/- 8.7 mV versus -7.1 +/- 5.1 mV, p < 0.01). The effects of amiloride on PD were reduced during exercise compared with rest in patients with CF (+15.8 +/- 9.5 mV versus +26.1 +/- 11.0 mV, p < 0.01) and control subjects (+5.8 +/- 4.8 mV versus +10.0 +/- 3.1 mV, p < 0.01). There was no effect of exercise on chloride conductance in patients with CF and control subjects. We conclude that moderate intensity exercise partially blocks the amiloride-sensitive sodium conductance in the respiratory epithelium. The inhibition of luminal sodium conductance could increase water content of the mucus in the CF lung during exercise and may, in part, explain the beneficial effects of exercise in patients with CF. PMID- 11500349 TI - Clinical importance of Alternaria exposure in children. AB - The fungus Alternaria is known to be allergenic and is one of the most common fungi worldwide. We investigated the extent to which exposure to Alternaria increases the severity of asthma. We undertook a prospective cohort study in Australia of 399 school children who had positive skin tests to one or more aeroallergens. Airway responsiveness to histamine, wheeze, and bronchodilator use in 1 mo was measured five times between 1997 and 1999. Airway hyperresponsiveness was defined as PD(20)FEV(1) = 3.9 micromol histamine. Airborne concentrations of Alternaria spores were measured throughout the study, and mean daily concentrations over 1 mo ranged from 2.2 to 307.7 spores/m(3) of ambient air. Using generalized estimating equations, we found that airway responsiveness, wheeze, and bronchodilator use increased significantly in association with increased spore concentrations and that the increase in airway responsiveness was greater in children sensitized to Alternaria than in other children (p = 0.01). The odds ratio for airway hyperresponsiveness in children sensitized to Alternaria was 1.26 (95% CI, 1.14 to 1.39) after an increase in mean exposure of 100 spore/m(3)/d over 1 mo. These results suggest that Alternaria allergens contribute to severe asthma in regions where exposure to the fungus is high. PMID- 11500348 TI - Bronchodilator responsiveness in normal infants and young children. AB - Several studies have demonstrated that normal infants exhibit bronchoconstriction after inhalation of nonspecific agonists and that the induced airway narrowing can be reversed by the inhalation of a beta-agonist. However, there are very limited data on baseline airway tone and the airway response to a beta-agonist in this subject population. The purpose of our study was to evaluate in normal infants baseline airway responsiveness to the inhaled beta-agonist, albuterol, using changes in maximal expiratory flows. Forty-one healthy infant volunteers with no history of respiratory disease or recurrent wheezing (ages 5.4 to 141.4 wk) were studied. Maximal expiratory flow- volume curves were obtained at baseline and 10 min after inhalation of albuterol (n = 28) or placebo (n = 13) using a metered-dose inhaler with a spacer. The mean percent change was significantly greater (p < 0.05) in the albuterol versus placebo group for FEV(0.5) (2.2% versus -1.5%), FEF(75%) (10.6% versus -3.1%), and FEF(85%) (12.9% versus 0.5%). Six of 28 albuterol-treated infants demonstrated increases in FEF(75%) greater than two standard deviations from the mean change in FEF(75%) seen in the placebo group. These infants were younger and more frequently exposed to maternal smoking during pregnancy. We conclude that normal healthy infants have overall levels of baseline airway tone that are similar to that reported in adults and older children; however, among the infants we evaluated the response to an inhaled bronchodilator was greatest in the youngest infants and in those exposed to tobacco smoking. KEYWORDS: airway responsiveness; asthma; tobacco smoke; infant pulmonary function; bronchodilator PMID- 11500350 TI - Diagnostic value of follow-up transbronchial lung biopsy after lung rejection. AB - Although transbronchial lung biopsy (TBBx) is widely acknowledged as the "gold standard" for diagnosis of acute rejection, controversy exists regarding the need to perform follow-up procedures. Over a 5-yr period, we performed 1,142 TBBx of which 173 were follow-up TBBx in 99 patients with pulmonary allograft rejection greater than or equal to International Society for Heart and Lung Transplantation (ISHLT) grade A(2) on initial TBBx. Rejection on the previous 173 TBBx was associated with lymphocytic bronchiolitis/bronchitis (LBB) > or = ISHLT grade B(2) in 82 patients and with cytomegalovirus (CMV) pneumonitis in 16 patients. Persistent rejection (> or = A(2)) was observed in 45 of 173 (26%) follow-up TBBx. Persistent B grade rejection (> or = B(2)) was present in 28 patients whereas new B grade rejection developed in 11 patients with > or = A(2) grade rejection. Rejection > or = B(2) was significantly (p < 0.05) associated with rejection > or = A(2). Fifteen follow-up TBBx showed new B grade rejection without signs of > or = A(2) rejection. A new diagnosis of CMV pneumonitis was made in 33 of 173 (19%). CMV pneumonitis occurred in 35 follow-up TBBx, four associated with > or = A(2) rejection and eight with > or = B(2) rejection. The overall incidence of bronchiolitis obliterans syndrome (BOS) in both groups was similar. Patients with persistent rejection on follow-up TBBx developed BOS at a median of 1.3 yr and median of 2.0 yr (p = not significant [NS]) posttransplantation. The practice of follow-up TBBx after rejection within 2 yr posttransplant is clinically useful as it provides valuable diagnostic information. PMID- 11500351 TI - Oral clearance and pathogenic oropharyngeal colonization in the elderly. AB - The elderly have an increased incidence of oropharyngeal colonization with respiratory pathogens, a well-known risk factor for the development of pneumonia. Changes in the oral milieu may occur secondary to decreased salivary production and abnormalities in swallowing. These abnormalities, common in the elderly, may result in impaired clearance of organisms, allowing pathogenic colonization. To test this hypothesis, we performed a prospective cross-sectional analysis of 75 elderly institutionalized patients and measured oral clearance using (99m)Tc human serum albumin (HSA) administered to the oropharynx. Oropharyngeal cultures, salivary cell populations, elastase activity, and clinical parameters were measured simultaneously. Retention of radiolabel ranged from 100% to 2.3% over 120 min of observation. Clearance in the oropharynx was significantly decreased in those patients who had oropharyngeal colonization with gram-negative bacilli (GNB), Staphylococcus aureus (SA), or yeast compared with those demonstrating normal flora by 95% confidence intervals. Decreased clearance was also seen in patients on antidepressants by 95% confidence levels. The absolute number of salivary lymphocytes/ml and buccal cells/ml was increased in colonized patients versus noncolonized persons (mean +/- SEM, 128 x 10(3) +/- 49 x 10(3), 25.4 +/- 11.6 x 10(3)). Elastase activity was elevated in patients who had GNB compared with patients without GNB (mean +/- SEM, 10.6 nM +/- 5.7, versus 2.2 nM +/- 1.2, p = 0.036). We conclude that a decrease in salivary clearance of potentially pathogenic organisms may be a major risk factor for the development of colonization in the elderly. PMID- 11500352 TI - Amplification of inflammation in emphysema and its association with latent adenoviral infection. AB - This study examines the hypothesis that the cigarette smoke-induced inflammatory process is amplified in severe emphysema and explores the association of this response with latent adenoviral infection. Lung tissue from patients with similar smoking histories and either no (n = 7), mild (n = 7), or severe emphysema (n = 7) was obtained by lung resection. Numbers of polymorphonuclear cells (PMN), macrophages, B cells, CD4, CD8 lymphocytes, and eosinophils present in tissue and airspaces and of epithelial cells expressing adenoviral E1A protein were determined using quantitative techniques. Severe emphysema was associated with an absolute increase in the total number of inflammatory cells in the lung tissue and airspaces. The computed tomography (CT) determined extent of lung destruction was related to the number of cells/m(2) surface area by R(2) values that ranged from 0.858 (CD8 cells) to 0.483 (B cells) in the tissue and 0.630 (CD4 cells) to 0.198 (B cells) in the airspaces. These changes were associated with a 5- to 40 fold increase in the number of alveolar epithelial cells expressing adenoviral E1A protein in mild and severe disease, respectively. We conclude that cigarette smoke-induced lung inflammation is amplified in severe emphysema and that latent expression of the adenoviral E1A protein expressed by alveolar epithelial cells influenced this amplification process. PMID- 11500353 TI - Airway smooth muscle cell proliferation is increased in asthma. AB - Increased airway smooth muscle (ASM) within the bronchial wall of asthmatic patients has been well documented and is likely to be the result of increased muscle proliferation. We have for the first time been able to culture ASM cells from asthmatic patients and to compare their proliferation rate with that of nonasthmatic patients. Asthmatic ASM cell cultures (n = 12) were established from explanted lungs and endobronchial biopsies. Nonasthmatic ASM cells (n = 10) were obtained from explanted tissue from patients with no airway disease, emphysema, carcinoma, and fibrosing alveolitis. Cell counts, tritiated thymidine incorporation, and cell cycle analysis were conducted over 7 d. Asthmatic ASM cell numbers at Days 3, 5, and 7 were significantly higher than corresponding values for nonasthmatic cells (p < 0.05). Tritiated thymidine incorporation was increased 3.2-fold in asthmatic cells compared with nonasthmatic cells within the first 24 h (p = 0.026). Flow cytometric analysis of DNA content on Days 1 and 2 revealed that a significantly greater percentage of asthmatic ASM cells were in the G2 + M phase (p < 0.05). This study shows for the first time that proliferation of ASM cells is increased in patients with asthma and provides evidence for an intrinsic abnormality in the ASM cell in this disease. KEYWORDS: asthma; human airway smooth muscle; cell culture; cell proliferation; hyperplasia PMID- 11500354 TI - Microgravity reduces sleep-disordered breathing in humans. AB - To understand the factors that alter sleep quality in space, we studied the effect of spaceflight on sleep-disordered breathing. We analyzed 77 8-h, full polysomnographic recordings (PSGs) from five healthy subjects before spaceflight, on four occasions per subject during either a 16- or 9-d space shuttle mission and shortly after return to earth. Microgravity was associated with a 55% reduction in the apnea-hypopnea index (AHI), which decreased from a preflight value of 8.3 +/- 1.6 to 3.4 +/- 0.8 events/h inflight. This reduction in AHI was accompanied by a virtual elimination of snoring, which fell from 16.5 +/- 3.0% of total sleep time preflight to 0.7 +/- 0.5% inflight. Electroencephalogram (EEG) arousals also decreased in microgravity (by 19%), and this decrease was almost entirely a consequence of the reduction in respiratory-related arousals, which fell from 5.5 +/- 1.2 arousals/h preflight to 1.8 +/- 0.6 inflight. Postflight there was a return to near or slightly above preflight levels in these variables. We conclude that sleep quality during spaceflight is not degraded by sleep disordered breathing. This is the first direct demonstration that gravity plays a dominant role in the generation of apneas, hypopneas, and snoring in healthy subjects. PMID- 11500355 TI - Structural alterations of gene complexes by cystic fibrosis sputum. AB - Gene complexes with optimal physicochemical characteristics for cystic fibrosis (CF) gene therapy in vitro may become inactive in vivo as a result of destruction upon interaction with CF mucus. Therefore, we examined in this study to what extent main sputum components (linear DNA, mucin, phosphatidylcholine, phosphatidylglycerol, and albumin) may disintegrate lipoplexes. We found that mixing linear DNA with lipoplexes, in concentration ratios as occurs in the mucus of patients with CF in clinical studies with lipoplexes, drastically altered the surface charge and size of our lipoplexes and resulted in the liberation of plasmid DNA from the lipoplexes. These concentration ratios occur in vivo when the DNA concentration in the sputum becomes > 2.7 mg/ml, a quite realistic concentration even in patients without acute exacerbations. Lipoplexes brought in contact with native CF sputa at clinically relevant concentration ratios dissociated when the DNA concentration in the sputa was > 2.7 mg/ml. However, when the linear DNA was degraded by recombinant human deoxyribonuclease I before lipoplexes were added, the linear DNA did not cause any dissociation of the lipoplexes. Addition of albumin and mucin to the lipoplexes in a clinically relevant concentration ratio changed the surface charge of the lipoplexes to negative, however, without release of plasmid DNA. Mucin, dipalmitoylglycerophosphocholine, and dipalmitoylglycerophosphoglycerol did not cause any change in lipoplex properties at clinically relevant concentration ratios. PMID- 11500356 TI - Effects of ventilation with different positive end-expiratory pressures on cytokine expression in the preterm lamb lung. AB - Ventilator-induced lung injury increases proinflammatory cytokines in the adult lung. We asked if positive end-expiratory pressure (PEEP) affects proinflammatory cytokine mRNA expression in the preterm lung. Preterm lambs at 129 +/- 3 d gestation were treated with 100 mg/kg recombinant human surfactant protein-C surfactant and ventilated for 2 or 7 h with 0, 4, or 7 cm H(2)O of PEEP. Unventilated fetal lambs were used as controls. Within 2 h of ventilation, alveolar total protein and activated neutrophils were increased and expression of mRNAs for the proinflammatory cytokines interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor-alpha (TNF-alpha) was increased in lung tissue of all ventilated animals relative to unventilated controls. Alveolar protein and neutrophils were higher for 0 and 7 PEEP animals than 4 PEEP animals. IL-1beta, IL-6, and IL-8 mRNAs were significantly elevated in animals ventilated with 0 PEEP compared with 4 PEEP. The percentage fractional area of collapsed alveoli was significantly higher for 0 PEEP compared with 4 and 7 PEEP groups. Mechanical ventilation increased the expression of proinflammatory mediators in surfactant treated preterm lungs and the use of 4 PEEP minimized this response. PMID- 11500357 TI - Pseudoasbestos bodies and fibers in bronchoalveolar lavage of refractory ceramic fiber users. AB - Refractory ceramic fibers (RCF) are widely used to replace asbestos in applications requiring high heat resistance. Ferruginous bodies mimicking asbestos bodies (ABs) have been detected in the lungs of RCF production workers. This raises the question about their presence in other occupational groups and whether "typical ABs" still reflect past asbestos exposures in all settings. An AB counting by phase-contrast light microscopy and a screening test by analytical electron microscopy were systematically performed on all bronchoalveolar lavage fluids (BALF) submitted to our laboratory in 1992 through 1997 (n = 1,800). When RCF were detected in electron microscopy, the structures considered as "typical ABs" were marked under light microscopy and prepared for further chemical and structural analysis. Pseudo-ABs on RCF were detected in samples from nine subjects (0.5%). All of them had worked either as foundry workers, steel workers, or welders. In these subjects, alumino-silicate fibers compatible with RCF accounted for 42% of the core fibers analyzed, other nonasbestos fibers for 28%, and asbestos fibers for 30%. ABs thus remain a valid marker of asbestos retention but attention must be paid to a possible occurrence of pseudo-asbestos bodies on RCF and other nonasbestos fibers in end-users of refractory fibers. PMID- 11500358 TI - New EMBO members' review: viral and bacterial proteins regulating apoptosis at the mitochondrial level. AB - Mitochondrial membrane permeabilization (MMP) is a critical step of several apoptotic pathways. Some infectious intracellular pathogens can regulate (induce or inhibit) apoptosis of their host cells at the mitochondrial level, by targeting proteins to mitochondrial membranes that either induce or inhibit MMP. Pathogen-encoded mitochondrion-targeted proteins may or may not show amino acid sequence homology to Bcl-2-like proteins. Among the Bcl-2-unrelated, mitochondrion-targeted proteins, several interact with the voltage-dependent anion channel (VDAC) or with the adenine nucleotide translocator (ANT). While VDAC-targeted proteins show homology to VDAC/porin, ANT-targeted proteins possess relatively short cationic binding domains, which may facilitate insertion into the negatively charged inner mitochondrial membrane. It may be speculated that such proteins employ pre-existing host-intrinsic mechanisms of MMP control. PMID- 11500359 TI - New EMBO members' review: actin cytoskeleton regulation through modulation of PI(4,5)P(2) rafts. AB - The phosphoinositide lipid PI(4,5)P(2) is now established as a key cofactor in signaling to the actin cytoskeleton and in vesicle trafficking. PI(4,5)P(2) accumulates at membrane rafts and promotes local co-recruitment and activation of specific signaling components at the cell membrane. PI(4,5)P(2) rafts may thus be platforms for local regulation of morphogenetic activity at the cell membrane. Raft PI(4,5)P(2) is regulated by lipid kinases (PI5-kinases) and lipid phosphatases (e.g. synaptojanin). In addition, GAP43-like proteins have recently emerged as a group of PI(4,5)P(2) raft-modulating proteins. These locally abundant proteins accumulate at inner leaflet plasmalemmal rafts where they bind to and co-distribute with PI(4,5)P(2), and promote actin cytoskeleton accumulation and dynamics. In keeping with their proposed role as positive modulators of PI(4,5)P(2) raft function, GAP43-like proteins confer competence for regulated morphogenetic activity on cells that express them. Their function has been investigated extensively in the nervous system, where their expression promotes neurite outgrowth, anatomical plasticity and nerve regeneration. Extrinsic signals and intrinsic factors may thus converge to modulate PI(4,5)P(2) rafts, upstream of regulated activity at the cell surface. PMID- 11500360 TI - New EMBO members' review: the double life of HMGB1 chromatin protein: architectural factor and extracellular signal. PMID- 11500361 TI - The Dictyostelium homologue of mammalian soluble adenylyl cyclase encodes a guanylyl cyclase. AB - A new Dictyostelium discoideum cyclase gene was identified that encodes a protein (sGC) with 35% similarity to mammalian soluble adenylyl cyclase (sAC). Gene disruption of sGC has no effect on adenylyl cyclase activity and results in a >10 fold reduction in guanylyl cyclase activity. The scg- null mutants show reduced chemotactic sensitivity and aggregate poorly under stringent conditions. With Mn(2+)/GTP as substrate, most of the sGC activity is soluble, but with the more physiological Mg(2+)/GTP the activity is detected in membranes and stimulated by GTPgammaS. Unexpectedly, orthologues of sGC and sAC are present in bacteria and vertebrates, but absent from Drosophila melanogaster, Caenorhabditis elegans, Arabidopsis thaliana and Saccharomyces cerevisiae. PMID- 11500362 TI - Eukaryotic initiation factor 2B: identification of multiple phosphorylation sites in the epsilon-subunit and their functions in vivo. AB - Eukaryotic initiation factor (eIF) 2B is a heteromeric guanine nucleotide exchange factor that plays an important role in regulating mRNA translation. Here we identify multiple phosphorylation sites in the largest, catalytic, subunit (epsilon) of mammalian eIF2B. These sites are phosphorylated by four different protein kinases. Two conserved sites (Ser712/713) are phosphorylated by casein kinase 2. They lie at the extreme C-terminus and are required for the interaction of eIF2Bepsilon with its substrate, eIF2, in vivo and for eIF2B activity in vitro. Glycogen synthase kinase 3 (GSK3) is responsible for phosphorylating Ser535. This regulatory phosphorylation event requires both the fourth site (Ser539) and a distal region, which acts to recruit GSK3 to eIF2Bepsilon in vivo. The fifth site, which lies outside the catalytic domain of eIF2Bepsilon, can be phosphorylated by casein kinase 1. All five sites are phosphorylated in the eIF2B complex in vivo. PMID- 11500363 TI - A novel method to identify protein kinase substrates: eEF2 kinase is phosphorylated and inhibited by SAPK4/p38delta. AB - We have developed a method of general application for identifying putative substrates of protein kinases in cell extracts. Using this procedure, we identified the physiological substrates of several mitogen-activated protein kinase kinases and an authentic substrate of stress-activated protein kinase (SAPK) 2a/p38. A 120 kDa protein was detected in skeletal muscle extracts that was phosphorylated rapidly by SAPK4/p38delta, but poorly by SAPK2/p38, SAPK3/p38gamma, SAPK1/JNK or extracellular signal-regulated kinase 2 (ERK2). It was purified and identified as eukaryotic elongation factor 2 kinase (eEF2K). SAPK4/p38delta phosphorylated eEF2K at Ser359 in vitro, causing its inactivation. eEF2K became phosphorylated at Ser359 and its substrate eEF2 became dephosphorylated (activated) when KB cells were exposed to anisomycin, an agonist that activates all SAPKs, including SAPK4/p38delta. The anisomycin-induced phosphorylation of Ser359 was unaffected by SB 203580, U0126 or rapamycin, and was prevented by overexpression of a catalytically inactive SAPK4/p38delta mutant, suggesting that SAPK4/p38delta may mediate the inhibition of eEF2K by this stress. The phosphorylation of eEF2K at Ser359 was also induced by insulin like growth factor-1. However, this was blocked by rapamycin, indicating that Ser359 is targeted by at least two signalling pathways. PMID- 11500364 TI - Regulation of elongation factor 2 kinase by p90(RSK1) and p70 S6 kinase. AB - Elongation factor 2 kinase (eEF2k) phosphorylates and inactivates eEF2. Insulin induces dephosphorylation of eEF2 and inactivation of eEF2 kinase, and these effects are blocked by rapamycin, which inhibits the mammalian target of rapamycin, mTOR. However, the signalling mechanisms underlying these effects are unknown. Regulation of eEF2 phosphorylation and eEF2k activity is lost in cells in which phosphoinositide-dependent kinase 1 (PDK1) has been genetically knocked out. This is not due to loss of mTOR function since phosphorylation of another target of mTOR, initiation factor 4E-binding protein 1, is not defective. PDK1 is required for activation of members of the AGC kinase family; we show that two such kinases, p70 S6 kinase (regulated via mTOR) and p90(RSK1) (activated by Erk), phosphorylate eEF2k at a conserved serine and inhibit its activity. In response to insulin-like growth factor 1, which activates p70 S6 kinase but not Erk, regulation of eEF2 is blocked by rapamycin. In contrast, regulation of eEF2 by stimuli that activate Erk is insensitive to rapamycin, but blocked by inhibitors of MEK/Erk signalling, consistent with the involvement of p90(RSK1). PMID- 11500365 TI - The PIF-binding pocket in PDK1 is essential for activation of S6K and SGK, but not PKB. AB - PKB/Akt, S6K1 and SGK are related protein kinases activated in a PI 3-kinase dependent manner in response to insulin/growth factors signalling. Activation entails phosphorylation of these kinases at two residues, the T-loop and the hydrophobic motif. PDK1 activates S6K, SGK and PKB isoforms by phosphorylating these kinases at their T-loop. We demonstrate that a pocket in the kinase domain of PDK1, termed the 'PIF-binding pocket', plays a key role in mediating the interaction and phosphorylation of S6K1 and SGK1 at their T-loop motif by PDK1. Our data indicate that prior phosphorylation of S6K1 and SGK1 at their hydrophobic motif promotes their interaction with the PIF-binding pocket of PDK1 and their T-loop phosphorylation. Thus, the hydrophobic motif phosphorylation of S6K and SGK converts them into substrates that can be activated by PDK1. In contrast, the PIF-binding pocket of PDK1 is not required for the phosphorylation of PKBalpha by PDK1. The PIF-binding pocket represents a substrate recognition site on a protein kinase that is only required for the phosphorylation of a subset of its physiological substrates. PMID- 11500366 TI - X-ray structure of junctional adhesion molecule: structural basis for homophilic adhesion via a novel dimerization motif. AB - Junctional adhesion molecules (JAMs) are a family of immunoglobulin-like single span transmembrane molecules that are expressed in endothelial cells, epithelial cells, leukocytes and myocardia. JAM has been suggested to contribute to the adhesive function of tight junctions and to regulate leukocyte trans migration. We describe the crystal structure of the recombinant extracellular part of mouse JAM (rsJAM) at 2.5 A resolution. rsJAM consists of two immunoglobulin-like domains that are connected by a conformationally restrained short linker. Two rsJAM molecules form a U-shaped dimer with highly complementary interactions between the N-terminal domains. Two salt bridges are formed in a complementary manner by a novel dimerization motif, R(V,I,L)E, which is essential for the formation of rsJAM dimers in solution and common to the known members of the JAM family. Based on the crystal packing and studies with mutant rsJAM, we propose a model for homophilic adhesion of JAM. In this model, U-shaped JAM dimers are oriented in cis on the cell surface and form a two-dimensional network by trans interactions of their N-terminal domains with JAM dimers from an opposite cell surface. PMID- 11500367 TI - Solution structure of the constant region of nuclear envelope protein LAP2 reveals two LEM-domain structures: one binds BAF and the other binds DNA. AB - The nuclear envelope proteins LAP2, emerin and MAN1 share a conserved approximately 40-residue 'LEM' motif. Loss of emerin causes Emery-Dreifuss muscular dystrophy. We have solved the solution NMR structure of the constant region of human LAP2 (residues 1-168). Human LAP2(1-168) has two structurally independent, non-interacting domains located at residues 1-50 ('LAP2-N') and residues 111-152 (LEM-domain), connected by an approximately 60-residue flexible linker. The two domains are structurally homologous, comprising a helical turn followed by two helices connected by an 11-12-residue loop. This motif is shared by subdomains of T4 endonuclease VII and transcription factor rho, despite negligible (< or =15%) sequence identity. NMR chemical shift mapping demonstrated that the LEM-domain binds BAF (barrier-to-autointegration factor), whereas LAP2-N binds DNA. Both binding surfaces comprise helix 1, the N-terminus of helix 2 and the inter-helical loop. Binding selectivity is determined by the nature of the surface residues in these binding sites, which are predominantly positively charged for LAP2-N and hydrophobic for the LEM-domain. Thus, LEM and LEM-like motifs form a common structure that evolution has customized for binding to BAF or DNA. PMID- 11500368 TI - Projection structure and molecular architecture of OxlT, a bacterial membrane transporter. AB - The major facilitator superfamily (MFS) represents the largest collection of evolutionarily related members within the class of membrane 'carrier' proteins. OxlT, a representative example of the MFS, is an oxalate-transporting membrane protein in Oxalobacter formigenes. From an electron crystallographic analysis of two-dimensional crystals of OxlT, we have determined the projection structure of this membrane transporter. The projection map at 6 A resolution indicates the presence of 12 transmembrane helices in each monomer of OxlT, with one set of six helices related to the other set by an approximate internal two-fold axis. The projection map reveals the existence of a central cavity, which we propose to be part of the pathway of oxalate transport. By combining information from the projection map with related biochemical data, we present probable models for the architectural arrangement of transmembrane helices in this protein superfamily. PMID- 11500369 TI - Functional relevance of the disulfide-linked complex of the N-terminal PDZ domain of InaD with NorpA. AB - In Drosophila, phototransduction is mediated by G(q)-activation of phospholipase C and is a well studied model system for understanding the kinetics of signal initiation, propagation and termination controlled by G proteins. The proper intracellular targeting and spatial arrangement of most proteins involved in fly phototransduction require the multi-domain scaffolding protein InaD, composed almost entirely of five PDZ domains, which independently bind various proteins including NorpA, the relevant phospho lipase C-beta isozyme. We have determined the crystal structure of the N-terminal PDZ domain of InaD bound to a peptide corresponding to the C-terminus of NorpA to 1.8 A resolution. The structure highlights an intermolecular disulfide bond necessary for high affinity interaction as determined by both in vitro and in vivo studies. Since other proteins also possess similar, cysteine-containing consensus sequences for binding PDZ domains, this disulfide-mediated 'dock-and-lock' interaction of PDZ domains with their ligands may be a relatively ubiquitous mode of coordinating signaling pathways. PMID- 11500370 TI - Cic1, an adaptor protein specifically linking the 26S proteasome to its substrate, the SCF component Cdc4. AB - In eukaryotes, the ubiquitin-proteasome system plays a major role in selective protein breakdown for cellular regulation. Here we report the discovery of a new essential component of this degradation machinery. We found the Saccharomyces cerevisiae protein Cic1 attached to 26S proteasomes playing a crucial role in substrate specificity for proteasomal destruction. Whereas degradation of short lived test proteins is not affected, cic1 mutants stabilize the F-box proteins Cdc4 and Grr1, substrate recognition subunits of the SCF complex. Cic1 interacts in vitro and in vivo with Cdc4, suggesting a function as a new kind of substrate recruiting factor or adaptor associated with the proteasome. PMID- 11500371 TI - Studies on influenza haemagglutinin fusion peptide mutants generated by reverse genetics. AB - Influenza haemagglutinin (HA) is responsible for fusing viral and endosomal membranes during virus entry. In this process, conformational changes in the HA relocate the HA(2) N-terminal 'fusion peptide' to interact with the target membrane. The highly conserved HA fusion peptide shares composition and sequence features with functionally analogous regions of other viral fusion proteins, including the presence and distribution of glycines and large side-chain hydrophobic residues. HAs with mutations in the fusion peptide were expressed using vaccinia virus recombinants to examine the requirement for fusion of specific hydrophobic residues and the significance of glycine spacing. Mutant HAs were also incorporated into infectious influenza viruses for analysis of their effects on infectivity and replication. In most cases alanine, but not glycine substitutions for the large hydrophobic residues, yielded fusion-competent HAs and infectious viruses, suggesting that the conserved spacing of glycines may be structurally significant. When viruses containing alanine substitutions for large hydrophobic residues were passaged, pseudoreversion to valine was observed, indicating a preference for large hydrophobic residues at specific positions. Viruses were also obtained with serine, leucine or phenylalanine as the N terminal residue, but these replicated to significantly lower levels than wild type virus with glycine at this position. PMID- 11500372 TI - HBV infection of cell culture: evidence for multivalent and cooperative attachment. AB - Hepadnaviruses do not infect cultured cells, therefore our knowledge of the mechanism of the early stages of virus-cell interaction is rather poor. In this study, we show that dimethylsulfoxide (DMSO)-treated HepG2 hepatoblastoma cells are infected efficiently by serum-derived hepatitis B virus (HBV) as monitored by viral gene expression and replication markers. To measure virus attachment, a variety of HBV surface proteins (HBsAgs) were conjugated to polystyrene beads and their capacity to attach cells was visualized and quantified by light microscopy at a single-cell resolution. Remarkably, DMSO increases the attachment efficiency by >200-fold. We further identify the QLDPAF sequence within preS1 as the receptor-binding viral domain epitope. Interestingly, a similar sequence is shared by several cellular, bacterial and viral proteins involved in cell adhesion, attachment and fusion. We also found that the small HBsAg contains a secondary attachment site that recognizes a distinct receptor on the cell membrane. Furthermore, we provide evidence in support of multivalent HBV attachment with synergistic interplay. Our data depict a mechanistic view of virus attachment and ingestion. PMID- 11500373 TI - Hes1 and Hes3 regulate maintenance of the isthmic organizer and development of the mid/hindbrain. AB - The isthmic organizer, which is located at the midbrain-hindbrain boundary, plays an essential role in development of the midbrain and anterior hindbrain. It has been shown that homeobox genes regulate establishment of the isthmic organizer, but the mechanism by which the organizer is maintained is not well understood. Here, we found that, in mice doubly mutant for the basic helix-loop-helix genes Hes1 and Hes3, the midbrain and anterior hindbrain structures are missing without any significant cell death. In these mutants, the isthmic organizer cells prematurely differentiate into neurons and terminate expression of secreting molecules such as Fgf8 and Wnt1 and the paired box genes Pax2/5, all of which are essential for the isthmic organizer function. These results indicate that Hes1 and Hes3 prevent premature differentiation and maintain the organizer activity of the isthmic cells, thereby regulating the development of the midbrain and anterior hindbrain. PMID- 11500374 TI - Identification of a mammalian H(+)-myo-inositol symporter expressed predominantly in the brain. AB - Inositol and its phosphorylated derivatives play a major role in brain function, either as osmolytes, second messengers or regulators of vesicle endo- and exocytosis. Here we describe the identification and functional characterization of a novel H(+)-myo- inositol co-transporter, HMIT, expressed predominantly in the brain. HMIT cDNA encodes a 618 amino acid polypeptide with 12 predicted transmembrane domains. Functional expression of HMIT in Xenopus oocytes showed that transport activity was specific for myo-inositol and related stereoisomers with a Michaelis-Menten constant of approximately 100 microM, and that transport activity was strongly stimulated by decreasing pH. Electrophysiological measurements revealed that transport was electrogenic with a maximal transport activity reached at pH 5.0. In rat brain membrane preparations, HMIT appeared as a 75-90 kDa protein that could be converted to a 67 kDa band upon enzymatic deglycosylation. Immunofluorescence microscopy analysis showed HMIT expression in glial cells and some neurons. These data provide the first characterization of a mammalian H(+)-coupled myo- inositol transporter. Predominant central expression of HMIT suggests that it has a key role in the control of myo-inositol brain metabolism. PMID- 11500375 TI - FANCC interacts with Hsp70 to protect hematopoietic cells from IFN-gamma/TNF alpha-mediated cytotoxicity. AB - The Fanconi anemia (FA) complementation group C gene product (FANCC) functions to protect hematopoietic cells from cytotoxicity induced by interferon-gamma (IFN gamma), tumor necrosis factor-alpha (TNF-alpha) and double-stranded RNA (dsRNA). Because apoptotic responses of mutant FA-C cells involve activation of interferon inducible, dsRNA-dependent protein kinase PKR, we sought to identify FANCC binding cofactors that may modulate PKR activation. We identified the molecular chaperone Hsp70 as an interacting partner of FANCC in lymphoblasts and HeLa cells using 'pull-down' and co-immunoprecipitation experiments. In vitro binding assays showed that the association of FANCC and Hsp70 involves the ATPase domain of Hsp70 and the central 320 residues of FANCC, and that both Hsp40 and ATP/ADP are required. In whole cells, Hsp70-FANCC binding and protection from IFN-gamma/TNF alpha-induced cytotoxicity were blocked by alanine mutations located in a conserved motif within the Hsp70-interacting domain of FANCC. We therefore conclude that FANCC acts in concert with Hsp70 to prevent apoptosis in hematopoietic cells exposed to IFN-gamma and TNF-alpha. PMID- 11500376 TI - RasC is required for optimal activation of adenylyl cyclase and Akt/PKB during aggregation. AB - Disruption of Dictyostelium rasC, encoding a Ras subfamily protein, generated cells incapable of aggregation. While rasC expression is enriched in a cell type specific manner during post-aggregative development, the defect in rasC(-) cells is restricted to aggregation and fully corrected by application of exogenous cAMP pulses. cAMP is not produced in rasC(-) cells stimulated by 2'-deoxy-cAMP, but is produced in response to GTPgammaS in cell lysates, indicating that G-protein coupled cAMP receptor activation of adenylyl cyclase is regulated by RasC. However, cAMP-induced ERK2 phosphorylation is unaffected in rasC(-) cells, indicating that RasC is not an upstream activator of the mitogen-activated protein kinase required for cAMP relay. rasC(-) cells also exhibit reduced chemotaxis to cAMP during early development and delayed response to periodic cAMP stimuli produced by wild-type cells in chimeric mixtures. Furthermore, cAMP induced Akt/PKB phosphorylation through a phosphatidylinositide 3-kinase (PI3K) dependent pathway is dramatically reduced in rasC(-) cells, suggesting that G protein-coupled serpentine receptor activation of PI3K is regulated by RasC. Cells lacking the RasGEF, AleA, exhibit similar defects as rasC(-) cells, suggesting that AleA may activate RasC. PMID- 11500377 TI - Negative regulation of the Wnt-beta-catenin pathway by the transcriptional repressor HBP1. AB - In certain cancers, constitutive Wnt signaling results from mutation in one or more pathway components. The result is the accumulation and nuclear localization of beta-catenin, which interacts with the lymphoid enhancer factor-1 (LEF)/T-cell factor (TCF) family of HMG-box transcription factors, which activate important growth regulatory genes, including cyclin D1 and c-myc. As exemplified by APC and axin, the negative regulation of beta-catenin is important for tumor suppression. Another potential mode of negative regulation is transcriptional repression of cyclin D1 and other Wnt target genes. In mammals, the transcriptional repressors in the Wnt pathway are not well defined. We have previously identified HBP1 as an HMG-box repressor and a cell cycle inhibitor. Here, we show that HBP1 is a repressor of the cyclin D1 gene and inhibits the Wnt signaling pathway. The inhibition of Wnt signaling and growth requires a common domain of HBP1. The apparent mechanism is an inhibition of TCF/LEF DNA binding through a physical interaction with HBP1. These data suggest that the suppression of Wnt signaling by HBP1 may be a mechanism to prevent inappropriate proliferation. PMID- 11500378 TI - Transcription of chromosomal rRNA genes by both RNA polymerase I and II in yeast uaf30 mutants lacking the 30 kDa subunit of transcription factor UAF. AB - UAF, a yeast RNA polymerase I transcription factor, contains Rrn5p, Rrn9p, Rrn10p, histones H3 and H4, and uncharacterized protein p30. Mutants defective in RRN5, RRN9 or RRN10 are unable to transcribe rDNA by polymerase I and grow extremely slowly, but give rise to variants able to grow by transcribing chromosomal rDNA by polymerase II. Thus, UAF functions as both an activator of polymerase I and a silencer of polymerase II for rDNA transcription. We have now identified the gene for subunit p30. This gene, UAF30, is not essential for growth, but its deletion decreases the cellular growth rate. Remarkably, the deletion mutants use both polymerase I and II for rDNA transcription, indicating that the silencer function of UAF is impaired, even though rDNA transcription by polymerase I is still occurring. A UAF complex isolated from the uaf30 deletion mutant was found to retain the in vitro polymerase I activator function to a large extent. Thus, Uaf30p plays only a minor role in its activator function. Possible reasons for slow growth caused by uaf30 mutations are discussed. PMID- 11500379 TI - Sir2p exists in two nucleosome-binding complexes with distinct deacetylase activities. AB - The absolute requirement for the histone deacetylase activity of Sir2p in silencing coupled with the conservation of Sir2p-like proteins in larger eukaryotes suggests that this molecule plays an important role in gene regulation in all organisms. Here we report the purification and characterization of two Sir2p-containing protein complexes; one of which contains Sir4p and the other Net1p. The Sir4p-containing complex has an NAD-dependent histone deacetylase activity, while the Net1p-containing complex possesses deacetylase activity but only weak NAD-dependent histone deacetylase activity. Finally, we demonstrate that the Sir2p-containing complexes bind nucleosomes efficiently and partially restrict accessibility of the linker DNA to enzymatic probes. PMID- 11500380 TI - A novel U2 and U11/U12 snRNP protein that associates with the pre-mRNA branch site. AB - Previous UV cross-linking studies demonstrated that, upon integration of the U2 snRNP into the spliceosome, a 14 kDa protein (p14) interacts directly with the branch adenosine, the nucleophile for the first transesterification step of splicing. We have identified the cDNA encoding this protein by microsequencing a 14 kDa protein isolated from U2-type spliceosomes. This protein contains an RNA recognition motif and is highly conserved across species. Antibodies raised against this cDNA-encoded protein precipitated the 14 kDa protein cross-linked to the branch adenosine, confirming the identity of the p14 cDNA. A combination of immunoblotting, protein microsequencing and immunoprecipitation revealed that p14 is a component of both 17S U2 and 18S U11/U12 snRNPs, suggesting that it contributes to the interaction of these snRNPs with the branch sites of U2- and U12-type pre-mRNAs, respectively. p14 was also shown to be a subunit of the heteromeric splicing factor SF3b and to interact directly with SF3b155. Immuno precipitations indicated that p14 is present in U12-type spliceosomes, consistent with the idea that branch point selection is similar in the major and minor spliceosomes. PMID- 11500381 TI - PML RING suppresses oncogenic transformation by reducing the affinity of eIF4E for mRNA. AB - The promyelocytic leukemia protein PML is organized into nuclear bodies which mediate suppression of oncogenic transformation and of growth. The biochemical functions of PML bodies are unknown, despite their involvement in several human disorders. We demonstrate that eukaryotic initiation factor 4E (eIF4E) directly binds the PML RING, a domain required for association with bodies and for suppression of transformation. Nuclear eIF4E functions in nucleocytoplasmic transport of a subset of transcripts including Cyclin D1. Present studies indicate that some PML requires the evolutionarily older eIF4E protein for association with nuclear bodies. Further more, PML RING modulates eIF4E activity by drastically reducing its affinity for its substrate, 5' m(7)G cap of mRNA. We demonstrate that eIF4E requires cap binding for transport of Cyclin D1 mRNA and subsequent transformation activity. Additionally, PML reduces the affinity of eIF4E for m(7)G mRNA cap, causing a reduction in Cyclin D1 protein levels and consequent transformation inhibition. PML is the first factor shown to modulate nuclear eIF4E function. These findings provide the first biochemical framework for understanding the transformation suppression activity of PML. PMID- 11500382 TI - Translation initiation factor IF3: two domains, five functions, one mechanism? AB - Initiation factor IF3 contains two domains separated by a flexible linker. While the isolated N-domain displayed neither affinity for ribosomes nor a detectable function, the isolated C-domain, added in amounts compensating for its reduced affinity for 30S subunits, performed all activities of intact IF3, namely: (i) dissociation of 70S ribosomes; (ii) shift of 30S-bound mRNA from 'stand-by' to 'P decoding' site; (iii) dissociation of 30S-poly(U)-NacPhe-tRNA pseudo- initiation complexes; (iv) dissociation of fMet-tRNA from initiation complexes containing mRNA with the non-canonical initiation triplet AUU (AUUmRNA); (v) stimulation of mRNA translation regardless of its start codon and inhibition of AUUmRNA translation at high IF3C/ribosome ratios. These results indicate that while IF3 performs all its functions through a C-domain-30S interaction, the N-domain function is to provide additional binding energy so that its fluctuating interaction with the 30S subunit can modulate the thermodynamic stability of the 30S-IF3 complex and IF3 recycling. The localization of IF3C far away from the decoding site and anticodon stem-loop of P-site-bound tRNA indicates that the IF3 fidelity function does not entail its direct contact with these structures. PMID- 11500383 TI - The intrinsic hypermutability of antibody heavy and light chain genes decays exponentially. AB - Somatic hypermutation, essential for the affinity maturation of antibodies, is restricted to a small segment of DNA. The upstream boundary is sharp and is probably related to transcription initiation. However, for reasons unknown, the hypermutation domain does not encompass the whole transcription unit, notably the C-region exon. Since analysis of the downstream decay of hypermutation is obscured by sequence-dependent hot and cold spots, we describe a strategy to minimize these fluctuations by computing mutations of different sequences located at similar distances from the promoter. We pool large databases of mutated heavy and light chains and analyse the decay of mutation frequencies. We define an intrinsic decay of probability of mutation that is remarkably similar for heavy and light chains, faster than anticipated and consistent with an exponential fit. Indeed, quite apart from hot spots, the intrinsic probability of mutation at CDR1 can be almost twice that of CDR3. The analysis has mechanistic implications for current and future models of hypermutation. PMID- 11500384 TI - Mechanistic aspects of DnaA-RepA interaction as revealed by yeast forward and reverse two-hybrid analysis. AB - Using yeast forward and reverse two-hybrid analysis and biochemical techniques, we present novel and definitive in vivo and in vitro evidence that both the N terminal domain I and C-terminal domain IV of the host-encoded DnaA initiator protein of Escherichia coli interact physically with plasmid-encoded RepA initiator of pSC101. The N-terminal, but not the C-terminal, region of RepA interacted with DnaA in vitro. These protein-protein interactions are critical for two very early steps of replication initiation, namely origin unwinding and helicase loading. Neither domain I nor IV of DnaA could individually collaborate with RepA to promote pSC101 replication. However, when the two domains are co expressed within a common cell milieu and allowed to associate non-covalently with each other via a pair of leucine zippers, replication of the plasmid was supported in vivo. Thus, the result shows that physical tethering, either non covalent or covalent, of domain I and IV of DnaA and interaction of both domains with RepA, are critical for replication initiation. The results also provide the molecular basis for a novel, potential, replication-based bacterial two-hybrid system. PMID- 11500385 TI - Human origin recognition complex binds to the region of the latent origin of DNA replication of Epstein-Barr virus. AB - Epstein-Barr virus (EBV) replicates in its latent phase once per cell cycle in proliferating B cells. The latent origin of DNA replication, oriP, supports replication and stable maintenance of the EBV genome. OriP comprises two essential elements: the dyad symmetry (DS) and the family of repeats (FR), both containing clusters of binding sites for the transactivator EBNA1. The DS element appears to be the functional replicator. It is not yet understood how oriP dependent replication is integrated into the cell cycle and how EBNA1 acts at the molecular level. Using chromatin immunoprecipitation experiments, we show that the human origin recognition complex (hsORC) binds at or near the DS element. The association of hsORC with oriP depends on the DS element. Deletion of this element not only abolishes hsORC binding but also reduces replication initiation at oriP to background level. Co-immunoprecipitation experiments indicate that EBNA1 is associated with hsORC in vivo. These results indicate that oriP might use the same cellular initiation factors that regulate chromosomal replication, and that EBNA1 may be involved in recruiting hsORC to oriP. PMID- 11500386 TI - CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells. AB - CENP-H has recently been discovered as a constitutive component of the centromere that co-localizes with CENP-A and CENP-C throughout the cell cycle. The precise function, however, remains poorly understood. We examined the role of CENP-H in centromere function and assembly by generating a conditional loss-of-function mutant in the chicken DT40 cell line. In the absence of CENP-H, cell cycle arrest at metaphase, consistent with loss of centromere function, was observed. Immunocytochemical analysis of the CENP-H-deficient cells demonstrated that CENP H is necessary for CENP-C, but not CENP-A, localization to the centromere. These findings indicate that centromere assembly in vertebrate cells proceeds in a hierarchical manner in which localization of the centromere-specific histone CENP A is an early event that occurs independently of CENP-C and CENP-H. PMID- 11500387 TI - c-Fos/activator protein-1 transactivates wee1 kinase at G(1)/S to inhibit premature mitosis in antigen-specific Th1 cells. AB - M-phase promoting factor is a complex of cdc2 and cyclin B that is regulated positively by cdc25 phosphatase and negatively by wee1 kinase. We isolated the wee1 gene promoter and found that it contains one AP-1 binding motif and is directly activated by the immediate early gene product c-Fos at cellular G(1)/S phase. In antigen-specific Th1 cells stimulated by antigen, transactivation of the c-fos and wee1 kinase genes occurred sequentially at G(1)/S, and the substrate of wee1 kinase, cdc2-Tyr15, was subsequently phosphorylated at late G(1)/S. Under prolonged expression of the c-fos gene, however, the amount of wee1 kinase was increased and its target cdc2 molecule was constitutively phosphorylated on its tyrosine residue, where Th1 cells went into aberrant mitosis. Thus, an immediate early gene product, c-Fos/AP-1, directly transactivates the wee1 kinase gene at G(1)/S. The transient increase in c-fos and wee1 kinase genes is likely to be responsible for preventing premature mitosis while the cells remain in the G(1)/S phase of the cell cycle. PMID- 11500388 TI - Escherichia coli translocation at the blood-brain barrier. PMID- 11500389 TI - Selection of glutamate-rich protein long synthetic peptides for vaccine development: antigenicity and relationship with clinical protection and immunogenicity. AB - Antibodies against three long synthetic peptides (LSPs) derived from the glutamate-rich protein (GLURP) of Plasmodium falciparum were analyzed in three cohorts from Liberia, Ghana, and Senegal. Two overlapping LSPs, LR67 and LR68, are derived from the relatively conserved N-terminal nonrepeat region (R0), and the third, LR70, is derived from the R2 repeat region. A high prevalence of antibody responses to each LSP was observed in all three areas of endemic infection. Levels of cytophilic immunoglobulin G (IgG) antibodies against both GLURP regions were significantly correlated with protection from clinical P. falciparum malaria. Protected children from the Ghana cohort possessed predominantly IgG1 antibodies against the nonrepeat epitope and IgG3 antibodies against the repeat epitope. T-cell proliferation responses, studied in the cohort from Senegal, revealed that T-helper-cell epitopes were confined to the nonrepeat region. When used as immunogens, the LR67 and LR68 peptides elicited strong IgG responses in outbred mice and LR67 also induced antibodies in mice of different H 2 haplotypes, confirming the presence of T-helper-cell epitopes in these constructs. Mouse antipeptide antisera recognized parasite proteins as determined by immunofluorescence and immunoblotting. This indicates that synthetic peptides derived from relatively conserved epitopes of GLURP might serve as useful immunogens for vaccination against P. falciparum malaria. PMID- 11500390 TI - Predominance of serotype-specific mucosal antibody response in Shigella flexneri infected humans living in an area of endemicity. AB - The mucosal humoral immune response elicited following Shigella flexneri infection in patients living in Antananarivo districts (Madagascar Island) was evaluated by measuring the gut-derived, circulating immunoglobulin A (IgA) antibody-secreting cells (ASC) specific for the major bacterial antigen lipopolysaccharide (LPS). Fifty, 34, 11, and 5% of the S. flexneri-positive patients were infected with serotypes 2a, 1a, 4a, and 3a, respectively. The total number of IgA ASC in infected patients increased significantly, compared to the number in healthy controls, early after the onset of disease. The number of anti homologous LPS IgA ASC varied among individuals and peaked between days 5 and 10 after the onset of the disease. In the S. flexneri 1a- and 2a-infected patients, the level of IgA ASC cross-reactivity to heterologous S. flexneri serotypes was weak. These data indicate that S. flexneri 2a and 1a are the predominant strains responsible for shigellosis in this area of endemicity and that the anti-LPS antibody response following natural infection is mainly directed against serotype specific determinants. PMID- 11500391 TI - Involvement of mitogen-activated protein kinase pathways in Staphylococcus aureus invasion of normal osteoblasts. AB - Staphylococcus aureus invades osteoblasts and can persist in the intracellular environment. The present study examined the role of osteoblast mitogen-activated protein kinase (MAPK) pathways in bacterial invasion. S. aureus infection of normal human and mouse osteoblasts resulted in an increase in the phosphorylation of the extracellular signal-regulated protein kinases (ERK 1 and 2). This stimulation of ERK 1 and 2 correlated with the time course of S. aureus invasion, and bacterial adherence induced the MAPK pathway. ERK 1 and 2 phosphorylation was time and dose dependent and required active S. aureus gene expression for maximal induction. The nonpathogenic Staphylococcus carnosus was also able to induce ERK 1 and 2 phosphorylation, albeit at lower levels than S. aureus. Phosphorylation of the stress-activated protein kinases was increased in both infected human and mouse osteoblasts; however, the p38 MAPK pathway was not activated in response to S. aureus. Finally, the transcription factor c-Jun, but not Elk-1 or ATF-2, was phosphorylated in response to S. aureus infection. PMID- 11500392 TI - Recognition of Lewis x derivatives present on mucins by flagellar components of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa binds to human respiratory mucins by mechanisms involving flagellar component-receptor interactions. The adhesion of P. aeruginosa strain PAK is mediated by the flagellar cap protein, FliD, without the involvement of flagellin. Two distinct types of FliD proteins have been identified in P. aeruginosa: A type, found in strain PAK, and B type, found in strain PAO1. In the present work, studies performed with the P. aeruginosa B-type strain PAO1 indicate that both the FliD protein and the flagellin of this strain are involved in the binding to respiratory mucins. Using polyacrylamide-based fluorescent glycoconjugates in a flow cytometry assay, it was previously demonstrated that P. aeruginosa recognizes Le(x) (or Lewis x) derivatives found at the periphery of human respiratory mucins. The aim of the present work was therefore to determine whether these carbohydrate epitopes (or glycotopes) are receptors for FliD proteins and flagellin. The results obtained by both flow cytometry and a microplate adhesion assay indicate that the FliD protein of strain PAO1 is involved in the binding of glycoconjugates bearing Le(x) or sialyl-Le(x) determinants, while the binding of flagellin is restricted to the glycoconjugate bearing Le(x) glycotope. In contrast, the type A cap protein of P. aeruginosa strain PAK is not involved in the binding to glycoconjugates bearing Le(x), sialyl-Le(x), or sulfosialyl-Le(x) glycotopes. This study demonstrates a clear association between a specific Pseudomonas adhesin and a specific mucin glycotope and demonstrates that fine specificities exist in mucin recognition by P. aeruginosa. PMID- 11500393 TI - Cellular mechanisms that cause suppressed gamma interferon secretion in endotoxin tolerant mice. AB - Endotoxin (lipopolysaccharide [LPS]) tolerance is a state of altered immunity characterized, in part, by suppression of LPS-induced gamma interferon (IFN gamma) expression. However, the cellular mediators regulating LPS-induced production of IFN-gamma in normal mice and the effect of LPS tolerance on these mediators has not been well characterized. Our studies show that macrophage dysfunction is the primary factor causing suppressed IFN-gamma expression in LPS tolerant mice. Specifically, LPS-tolerant macrophages have a markedly impaired ability to induce IFN-gamma secretion by T cells and NK cells obtained from either control or LPS-tolerant mice. However, T cells and NK cells isolated from LPS-tolerant mice produce normal levels of IFN-gamma when cocultured with control macrophages or exogenous IFN-gamma-inducing factors. Assessment of important IFN gamma-regulating factors showed that interleukin-12 (IL-12) and costimulatory signals provided by IL-15, IL-18, and CD86 are largely responsible for LPS induced IFN-gamma expression in control mice. IL-10 is an inhibitor of IFN-gamma production in both the control and LPS-tolerant groups. Expression of IL-12 and the IL-12 receptor beta1 (IL-12Rbeta1) and IL-12Rbeta2 subunits are suppressed in the spleens of LPS-tolerant mice. LPS-tolerant splenocytes also exhibit decreased production of IL-15 and IL-15Ralpha. However, expression of IL-18 and the B7 proteins CD80 and CD86 are unchanged or increased compared to controls after induction of LPS tolerance. CD28, a major receptor for B7 proteins, is also increased in the spleens of LPS-tolerant mice. Expression of the inhibitory cytokine IL-10 and the IL-10R are sustained after induction of LPS tolerance. These data show that suppression of IFN-gamma production in LPS-tolerant mice is largely due to macrophage dysfunction and provide insight into the cellular alterations that occur in LPS tolerance. This study also better defines the factors that mediate LPS-induced IFN-gamma production in normal mice. PMID- 11500394 TI - CD4+ T helper 1 cells facilitate regression of murine Lyme carditis. AB - Murine Lyme borreliosis, caused by infection with the spirochete Borrelia burgdorferi, results in acute arthritis and carditis that regress as a result of B. burgdorferi-specific immune responses. B. burgdorferi-specific antibodies can attenuate arthritis in mice deficient in both B cells and T cells but have no effect on carditis. Because macrophages comprise the principal immune cell in carditis, T-cell responses that augment cell-mediated immunity may be important for carditis regression. To investigate this hypothesis, we examined the course of Lyme carditis in mice selectively deficient in B cells or alphabeta T cells. Our results show that carditis regresses in B-cell-deficient B10.A(k) mice but not in alphabeta T-cell-deficient mice, independently of the mouse strain background. Despite prominent macrophage infiltrates, hearts from B. burgdorferi infected alphabeta T-cell-deficient mice had less mRNA for tumor necrosis factor alpha as measured by reverse transcription-PCR compared to infected control mice. Anti-inflammatory cytokine mRNA levels were equivalent. Adoptive transfer of gamma interferon-secreting CD4+ T cells into infected alphabeta T-cell-deficient mice promoted carditis resolution. These results show that alphabeta T cells can promote resolution of murine Lyme carditis and are the first demonstration of a beneficial role for CD4+ T helper 1 cells in this disease. PMID- 11500395 TI - Potent stimulation of the innate immune system by a Leishmania brasiliensis recombinant protein. AB - The interaction of the innate immune system with the microbial world involves primarily two sets of molecules generally known as microbial pattern recognition receptors and microbial pattern recognition molecules, respectively. Examples of the former are the Toll receptors present particularly in macrophages and dendritic cells. Conversely, the microbial pattern recognition molecules are conserved protist homopolymers, such as bacterial lipopolysaccharides, lipoteichoic acids, peptidoglycans, glucans, mannans, unmethylated bacterial DNA, and double-strand viral RNA. However, for protists that lack most of these molecules, such as protozoans, the innate immune system must have evolved receptors that recognize other groups of microbial molecules. Here we present evidence that a highly purified protein encoded by a Leishmania brasiliensis gene may be one such molecule. This recombinant leishmanial molecule, a homologue of eukaryotic ribosomal elongation and initiation factor 4a (LeIF), strongly stimulates spleen cells from severe combined immunodeficient (SCID) mice to produce interleukin-12 (IL-12), IL-18, and high levels of gamma interferon. In addition, LeIF potentiates the cytotoxic activity of the NK cells of these animals. Because LeIF is a conserved molecule and because SCID mice lack T and B lymphocytes but have a normal innate immune system (normal reticuloendothelial system and NK cells), these results suggest that proteins may also be included as microbial pattern recognition molecules. The nature of the receptor involved in this innate recognition is unknown. However, it is possible to exclude the Toll receptor Tlr4 as a putative LeIF receptor because the gene encoding this receptor is defective in C3H/HeJ mice, the mouse strain used in the present studies. PMID- 11500396 TI - vsp gene expression by Giardia lamblia clone GS/M-83-H7 during antigenic variation in vivo and in vitro. AB - Giardia lamblia infections are associated with antigenic variation of the parasite, which is generated by a continuous change of the variant-specific surface proteins (VSPs). Many investigations on the process of antigenic variation were based on the use of G. lamblia clone GS/M-83-H7, which expresses VSP H7 as its major surface antigen. In the present study, mice were infected with the aforementioned clonal line to investigate vsp gene expression during the complex process of antigenic variation of the parasite. Trophozoites collected from the intestines of individual animals at different time points postinfection (p.i.) were analyzed directly for their vsp gene expression patterns, i.e., without cultivating the recovered parasites in vitro. Because few trophozoites were recovered at late time points p.i., a combined 5' rapid amplification of cDNA ends-reverse transcription-PCR approach was utilized. This allowed detection and subsequent sequence analysis of vsp gene transcripts upon generation of amplified cDNA analogues. The same PCR approach was applied for analysis of vsp gene expression in variants obtained after negative selection of axenic GS/M-83 H7 trophozoites by treatment with a cytotoxic, VSP H7-specific monoclonal antibody. In an overall view of the entire panel of in vivo- and in vitro-derived parasite populations, expression of 29 different vsp gene sequences could be demonstrated. In vivo antigenic variation of G. lamblia clone GS/M-83-H7 was shown to be a continuous process involving the consecutive appearance of relatively distinct sets of vsp transcripts. During the 42-day infection period investigated, this process activated at least 22 different vsp genes. Comparative molecular analyses of the amino acid level demonstrated that all cDNA segments identified encode structural elements typical of the terminal segment of Giardia VSP. The similarity of most of the GS/M-83-H7 VSP sequences identified in the present study supports previous suggestions that vsp gene diversification in G. lamblia is the result of ancestral gene duplication, mutation, and/or recombination events. PMID- 11500398 TI - Pulmonary and systemic host response to Streptococcus pneumoniae and Klebsiella pneumoniae bacteremia in normal and immunosuppressed mice. AB - Mortality related to bacteremic pneumonia remains high, and the role of sepsis in inflammation, pulmonary injury, and death remains unclear, mostly in leukopenic states. In the present study, the microbiology, histopathology, and host response to Streptococcus pneumoniae and Klebsiella pneumoniae infection were determined in an experimental model of bacteremia in immunocompetent and leukopenic mice. Leukocyte depletion by cyclophosphamide did not impair the early clearance of pneumococci from blood but facilitated growth in lungs. By contrast, klebsiellae rapidly grew in blood of leukopenic mice. These observations suggest that tissue based phagocytes and circulating leukocytes, respectively, play prominent roles in S. pneumoniae and K. pneumoniae eradication. The kinetics of leukocyte recruitment in lungs during S. pneumoniae bacteremia suggested early strong inflammation in immunocompetent mice that is associated with tumor necrosis factor alpha release and histological disorders, including cell debris and surfactant in alveolar spaces. Leukocyte depletion further stimulated pulmonary capillary leakage both in S. pneumoniae and K. pneumoniae bacteremia, which seemed attributable to bacterial virulence factors. Nitric oxide production did not differ significantly among groups. Leukopenia and low platelet counts characterized the late stage of bacteremia for both strains, but only K. pneumoniae altered renal function. Understanding the pathogenesis of bacteremia will help establish beneficial therapies for both sepsis and pneumonia. PMID- 11500397 TI - Borrelia burgdorferi RevA antigen is a surface-exposed outer membrane protein whose expression is regulated in response to environmental temperature and pH. AB - Borrelia burgdorferi, the causative agent of Lyme disease, produces RevA protein during the early stages of mammalian infection. B. burgdorferi apparently uses temperature as a cue to its location, producing proteins required for infection of warm-blooded animals at temperatures corresponding to host body temperature, but does not produce such virulence factors at cooler, ambient temperatures. We have observed that B. burgdorferi regulates expression of RevA in response to temperature, with the protein being synthesized by bacteria cultivated at 34 degrees C but not by those grown at 23 degrees C. Tissues encountered by B. burgdorferi during its infectious cycle vary in their pH values, and the level of RevA expression was also found to be dependent upon pH of the culture medium. The cellular localization of RevA was also analyzed. Borrelial inner and outer membranes were purified by isopycnic centrifugation, and membrane fractions were conclusively identified by immunoblot analysis using antibodies raised against the integral inner membrane protein MotB and outer membrane-associated Erp lipoproteins. Immunoblot analyses indicated that RevA is located in the B. burgdorferi outer membrane. These analyses also demonstrated that an earlier report (H. A. Bledsoe et al., Infect. Immun. 176:7447-7455, 1994) had misidentified such B. burgdorferi membrane fractions. RevA was further demonstrated to be exposed to the external environment, where it could facilitate interactions with host tissues. PMID- 11500400 TI - Kinetics of Bartonella birtlesii infection in experimentally infected mice and pathogenic effect on reproductive functions. AB - The kinetics of infection and the pathogenic effects on the reproductive function of laboratory mice infected with Bartonella birtlesii recovered from an Apodemus species are described. B. birtlesii infection, as determined by bacteremia, occurred in BALB/c mice inoculated intravenously. Inoculation with a low-dose inoculum (1.5 x 10(3) CFU) induced bacteremia in only 75% of the mice compared to all of the mice inoculated with higher doses (> or =1.5 x 10(4)). Mice became bacteremic for at least 5 weeks (range, 5 to 8 weeks) with a peak ranging from 2 x 10(3) to 10(5) CFU/ml of blood. The bacteremia level was significantly higher in virgin females than in males but the duration of bacteremia was similar. In mice infected before pregnancy (n = 20), fetal loss was evaluated by enumerating resorption and fetal death on day 18 of gestation. The fetal death and resorption percentage of infected mice was 36.3% versus 14.5% for controls (P < 0.0001). Fetal suffering was evaluated by weighing viable fetuses. The weight of viable fetuses was significantly lower for infected mice than for uninfected mice (P < 0.0002). Transplacental transmission of Bartonella was demonstrated since 76% of the fetal resorptions tested was culture positive for B. birtlesii. The histopathological analysis of the placentas of infected mice showed vascular lesions in the maternal placenta, which could explain the reproductive disorders observed. BALB/c mice appeared to be a useful model for studying Bartonella infection. This study provides the first evidence of reproductive disorders in mice experimentally infected with a Bartonella strain originating from a wild rodent. PMID- 11500399 TI - Role of trehalose dimycolate in recruitment of cells and modulation of production of cytokines and NO in tuberculosis. AB - Mice treated with viable Mycobacterium tuberculosis with no glycolipid trehalose dimycolate (TDM) on the outer cell wall (delipidated M. tuberculosis) by intraperitoneal or intratracheal inoculation presented an intense recruitment of polymorphonuclear cells into the peritoneal cavity and an acute inflammatory reaction in the lungs, respectively. In addition, lung lesions were resolved around the 32nd day after intratracheal inoculation. TDM-loaded biodegradable poly-DL-lactide-coglycolide microspheres as well as TDM-coated charcoal particles induced an intense inflammatory reaction. In addition, high levels of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha), IL-12, IL-10, gamma interferon (IFN-gamma), and IL-4 production were detected in lung cells, and nitric oxide (NO) production was high in culture supernatants of bronchoalveolar lavage cells. These in vivo data were confirmed by in vitro experiments using peritoneal macrophages cultured in the presence of TDM adsorbed onto coverslips. High levels of IFN-gamma, IL-6, TNF-alpha, IL-12, IL-10, and NO were detected in the culture supernatants. Our results suggest that TDM contributes to persistence of infection through production of cytokines, which are important for the recruitment of inflammatory cells and maintenance of a granulomatous reaction. In addition, our findings are important for a better understanding of the immunostimulatory activity of TDM and its possible use as an adjuvant in experiments using DNA vaccine or gene therapy against tuberculosis. PMID- 11500401 TI - Independent and coordinate effects of ADP-ribosyltransferase and GTPase activating activities of exoenzyme S on HT-29 epithelial cell function. AB - Type III-mediated translocation of exoenzyme S (ExoS) into HT-29 epithelial cells by Pseudomonas aeruginosa causes complex alterations in cell function, including inhibition of DNA synthesis, altered cytoskeletal structure, loss of readherence, microvillus effacement, and interruption of signal transduction. ExoS is a bifunctional protein having both GTPase-activating (GAP) and ADP ribosyltransferase (ADPRT) functional domains. Comparisons of alterations in HT 29 cell function caused by P. aeruginosa strains that translocate ExoS having GAP or ADPRT mutations allowed the independent and coordinate functions of the two activities to be assessed. An E381A ADPRT mutation revealed that ExoS ADPRT activity was required for effects of ExoS on DNA synthesis and long-term cell rounding. Conversely, the R146A GAP mutation appeared to have little impact on the cellular effects of ExoS. While transient cell rounding was detected following exposure to the E381A mutant, this rounding was eliminated by an E379A E381A ADPRT double mutation, implying that residual ADPRT activity, rather than GAP activity, was effecting transient cell rounding by the E381A mutant. To explore this possibility, E381A and R146A-E381A mutants were examined for their ability to ADP-ribosylate Ras in vitro or in vivo. While no ADP-ribosylation of Ras was detected by either mutant in vitro, both mutants were able to modify Ras when translocated by the bacteria, with the R146A-E381A mutant causing more efficient modification than the E381A mutant, in association with increased inhibition of DNA synthesis. Comparisons of Ras ADP-ribosylation by wild-type and E381A mutant ExoS by two-dimensional electrophoresis found the former to ADP ribosylate Ras at two sites, while the latter modified Ras only once. These studies draw attention to the key role of ExoS ADPRT activity in causing the effects of bacterially translocated ExoS on DNA synthesis and cell rounding. In addition, the studies provide insight into the enhancement of ExoS ADPRT activity within the eukaryotic cell microenvironment and into possible modulatory roles that the GAP and ADPRT domains might have on the function of each other. PMID- 11500402 TI - ADP-ribosylation of Rab5 by ExoS of Pseudomonas aeruginosa affects endocytosis. AB - Pseudomonas aeruginosa exoenzyme S (ExoS) is an ADP-ribosyltransferase that modifies low-molecular-weight GTPases. Here we studied the effect of Rab5 ADP ribosylation by ExoS on its cellular function, i.e., regulation of early endocytic events. Coculture of CHO cells with P. aeruginosa induced a marked decrease in horseradish peroxidase (HRP) uptake compared to noninfected cells, while coculture with a P. aeruginosa mutant strain that fails to produce ExoS did not lead to any change in HRP uptake. Microinjection of recombinant ExoS into Xenopus oocytes induced strong inhibition of basal HRP uptake by oocytes. Moreover, coinjection of recombinant ExoS with Rab5 abolished the typical stimulation of HRP uptake obtained after GTPase microinjection. Cytosols prepared from injected oocytes were used in an endosome-endosome fusion assay. Cytosol from ExoS-microinjected oocytes was ineffective in promoting endosome-endosome fusion. However, in these conditions, the addition of Rab5 to the assay led to fusion recovery. Finally, we found that the interaction of Rab5 with EEA1 was markedly diminished after Rab5 ADP-ribosylation by ExoS. PMID- 11500403 TI - Comparison of virulence of Legionella longbeachae strains in guinea pigs and U937 macrophage-like cells. AB - A guinea pig model of experimental legionellosis was established for assessment of virulence of isolates of Legionella longbeachae. The results showed that there were distinct virulence groupings of L. longbeachae serogroup 1 strains based on the severity of disease produced in this model. Statistical analysis of the animal model data suggests that Australian isolates of L. longbeachae may be inherently more virulent than non-Australian strains. Infection studies performed with U937 cells were consistent with the animal model studies and showed that isolates of this species were capable of multiplying within these phagocytic cells. Electron microscopy studies of infected lung tissue were also undertaken to determine the intracellular nature of L. longbeachae serogroup 1 infection. The data showed that phagosomes containing virulent L. longbeachae serogroup 1 appeared bloated, contained cellular debris and had an apparent rim of ribosomes while those containing avirulent L. longbeachae serogroup 1 were compact, clear and smooth. PMID- 11500404 TI - T-cell reactivity against streptococcal antigens in the periphery mirrors reactivity of heart-infiltrating T lymphocytes in rheumatic heart disease patients. AB - T-cell molecular mimicry between streptococcal and heart proteins has been proposed as the triggering factor leading to autoimmunity in rheumatic heart disease (RHD). We searched for immunodominant T-cell M5 epitopes among RHD patients with defined clinical outcomes and compared the T-cell reactivities of peripheral blood and intralesional T cells from patients with severe RHD. The role of HLA class II molecules in the presentation of M5 peptides was also evaluated. We studied the T-cell reactivity against M5 peptides and heart proteins on peripheral blood mononuclear cells (PBMC) from 74 RHD patients grouped according to the severity of disease, along with intralesional and peripheral T-cell clones from RHD patients. Peptides encompassing residues 1 to 25, 81 to 103, 125 to 139, and 163 to 177 were more frequently recognized by PBMC from RHD patients than by those from controls. The M5 peptide encompassing residues 81 to 96 [M5(81-96) peptide] was most frequently recognized by PBMC from HLA-DR7+ DR53+ patients with severe RHD, and 46.9% (15 of 32) and 43% (3 of 7) of heart-infiltrating and PBMC-derived peptide-reactive T-cell clones, respectively, recognized the M5(81-103) region. Heart proteins were recognized more frequently by PBMC from patients with severe RHD than by those from patients with mild RHD. The similar pattern of T-cell reactivity found with both peripheral blood and heart-infiltrating T cells is consistent with the migration of M-protein sensitized T cells to the heart tissue. Conversely, the presence of heart reactive T cells in the PBMC of patients with severe RHD also suggests a spillover of sensitized T cells from the heart lesion. PMID- 11500405 TI - Cellular and humoral immune responses and protection against schistosomes induced by a radiation-attenuated vaccine in chimpanzees. AB - The radiation-attenuated Schistosoma mansoni vaccine is highly effective in rodents and primates but has never been tested in humans, primarily for safety reasons. To strengthen its status as a paradigm for a human recombinant antigen vaccine, we have undertaken a small-scale vaccination and challenge experiment in chimpanzees (Pan troglodytes). Immunological, clinical, and parasitological parameters were measured in three animals after multiple vaccinations, together with three controls, during the acute and chronic stages of challenge infection up to chemotherapeutic cure. Vaccination induced a strong in vitro proliferative response and early gamma interferon production, but type 2 cytokines were dominant by the time of challenge. The controls showed little response to challenge infection before the acute stage of the disease, initiated by egg deposition. In contrast, the responses of vaccinated animals were muted throughout the challenge period. Vaccination also induced parasite-specific immunoglobulin M (IgM) and IgG, which reached high levels at the time of challenge, while in control animals levels did not rise markedly before egg deposition. The protective effects of vaccination were manifested as an amelioration of acute disease and overall morbidity, revealed by differences in gamma-glutamyl transferase level, leukocytosis, eosinophilia, and hematocrit. Moreover, vaccinated chimpanzees had a 46% lower level of circulating cathodic antigen and a 38% reduction in fecal egg output, compared to controls, during the chronic phase of infection. PMID- 11500406 TI - Ongoing horizontal and vertical transmission of virulence genes and papA alleles among Escherichia coli blood isolates from patients with diverse-source bacteremia. AB - The phylogenetic distributions of multiple putative virulence factors (VFs) and papA (P fimbrial structural subunit) alleles among 182 Escherichia coli blood isolates from patients with diverse-source bacteremia were defined. Phylogenetic correspondence among these strains, the E. coli Reference (ECOR) collection, and other collections of extraintestinal pathogenic E. coli (ExPEC) was assessed. Although among the 182 bacteremia isolates phylogenetic group B2 predominated, exhibited the greatest concentration of individual VFs, and contained the largest number of familiar virulent clones, other phylogenetic groups exhibited greater concentrations of certain VFs than did group B2 and included several additional virulent clones. Certain of the newly detected VF genes, e.g., fyuA (yersiniabactin; 76%) and focG (F1C fimbriae; 25%), were as prevalent or more prevalent than their more familiar traditional counterparts, e.g., iut (aerobactin; 57%) and sfaS (S fimbriae; 14%), thus possibly offering additional useful targets for preventive interventions. Considerable diversity of VF profiles was observed at every level within the phylogenetic tree, including even within individual lineages. This suggested that many different pathways can lead to extraintestinal virulence in E. coli and that the evolution of ExPEC, which involves extensive horizontal transmission of VFs and continuous remodeling of pathogenicity-associated islands, is a highly active, ongoing process. PMID- 11500407 TI - Structural and genetic analyses of O polysaccharide from Actinobacillus actinomycetemcomitans serotype f. AB - The oral bacterium Actinobacillus actinomycetemcomitans is implicated as a causative agent of localized juvenile periodontitis (LJP). A. actinomycetemcomitans is classified into five serotypes (a to e) corresponding to five structurally and antigenically distinct O polysaccharide (O-PS) components of their respective lipopolysaccharide molecules. Serotype b has been reported to be the dominant serotype isolated from LJP patients. We determined the lipopolysaccharide O-PS structure from A. actinomycetemcomitans CU1000, a strain isolated from a 13-year-old African-American female with LJP which had previously been classified as serotype b. The O-PS of strain CU1000 consisted of a trisaccharide repeating unit composed of L-rhamnose and 2-acetamido-2-deoxy-D galactose (molar ratio, 2:1) with the structure -->2)-alpha-L-Rhap-(1-3)-2-O (beta-D-GalpNAc)-alpha-L-Rhap-(1-->* O-PS from strain CU1000 was structurally and antigenically distinct from the O-PS molecules of the five known A. actinomycetemcomitans serotypes. Strain CU1000 was mutagenized with transposon IS903phikan, and three mutants that were deficient in O-PS synthesis were isolated. All three transposon insertions mapped to a single 1-kb region on the chromosome. The DNA sequence of a 13.1-kb region surrounding these transposon insertions contained a cluster of 14 open reading frames that was homologous to gene clusters responsible for the synthesis of A. actinomycetemcomitans serotype b, c, and e O-PS antigens. The CU1000 gene cluster contained two genes that were not present in serotype-specific O-PS antigen clusters of the other five known A. actinomycetemcomitans serotypes. These data indicate that strain CU1000 should be assigned to a new A. actinomycetemcomitans serotype, designated serotype f. A PCR assay using serotype-specific PCR primers showed that 3 out of 20 LJP patients surveyed (15%) harbored A. actinomycetemcomitans strains carrying the serotype f gene cluster. The finding of an A. actinomycetemcomitans serotype showing serological cross-reactivity with anti-serotype b-specific antiserum suggests that a reevaluation of strains previously classified as serotype b may be warranted. PMID- 11500408 TI - Characterization of an endoprotease (PrpL) encoded by a PvdS-regulated gene in Pseudomonas aeruginosa. AB - The expression of many virulence factors in Pseudomonas aeruginosa is dependent upon environmental conditions, including iron levels, oxygen, temperature, and osmolarity. The virulence of P. aeruginosa PAO1 is influenced by the iron- and oxygen-regulated gene encoding the alternative sigma factor PvdS, which is regulated through the ferric uptake regulator (Fur). We observed that overexpression of PvdS in strain PAO1 and a DeltapvdS::Gm mutant resulted in increased pyoverdine production and proteolytic activity compared to when PvdS was not overexpressed. To identify additional PvdS-regulated genes, we compared extracellular protein profiles from PAO1 and the DeltapvdS::Gm mutant grown under iron-deficient conditions. A protein present in culture supernatants from PAO1 but not in supernatants from DeltapvdS::Gm was investigated. Amino acid sequence analysis and examination of the genomic database of PAO1 revealed that the N terminus of this 27-kDa protein is identical to that of protease IV of P. aeruginosa strain PA103-29 and is homologous to an endoprotease produced by Lysobacter enzymogenes. In this study, the gene encoding an endoprotease was cloned from PAO1 and designated prpL (PvdS-regulated endoprotease, lysyl class). All (n = 41) but one of the strains of P. aeruginosa, including clinical and environmental isolates, examined carry prpL. Moreover, PrpL production among these strains was highly variable. Analysis of RNase protection assays identified the transcription initiation site of prpL and confirmed that its transcription is iron dependent. In the DeltapvdS::Gm mutant, the level of prpL transcription was iron independent and decreased relative to the level in PAO1. Furthermore, transcription of prpL was independent of PtxR, a PvdS-regulated protein. Finally, PrpL cleaves casein, lactoferrin, transferrin, elastin, and decorin and contributes to PAO1's ability to persist in a rat chronic pulmonary infection model . PMID- 11500409 TI - Roles of fructosyltransferase and levanase-sucrase of Actinomyces naeslundii in fructan and sucrose metabolism. AB - The ability of Actinomyces naeslundii to convert sucrose to extracellular homopolymers of fructose and to catabolize these types of polymers is suspected to be a virulence trait that contributes to the initiation and progression of dental caries and periodontal diseases. Previously, we reported on the isolation and characterization of the gene, ftf, encoding the fructosyltransferase (FTF) of A. naeslundii WVU45. Allelic exchange mutagenesis was used to inactivate ftf, revealing that FTF-deficient stains were completely devoid of the capacity to produce levan-type (beta2,6-linked) polysaccharides. A polyclonal antibody was raised to a histidine-tagged, purified A. naeslundii FTF, and the antibody was used to localize the enzyme in the supernatant fluid. A sensitive technique was developed to detect levan formation by proteins that had been separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the method was used to confirm that the levan-synthesizing activity of A. naeslundii existed predominantly in a cell-free form, that a small amount of the activity was cell associated, and that the ftf mutant was unable to produce levans. By using the nucleotide sequence of the levanase gene of a genospecies 2 A. naeslundii, formerly Actinomyces viscosus, a portion of a homologue of this gene (levJ) was amplified by PCR and inserted into a suicide vector, and the resulting construct was used to inactivate the levJ gene in the genospecies 1 strain WVU45. A variety of physiologic and biochemical studies were performed on the wild-type and LevJ deficient strains to demonstrate that (i) this enzyme was the dominant levanase and sucrase of A. naeslundii; (ii) that LevJ was inducible by growth in sucrose; (iii) that the LevJ activity was found predominantly (>90%) in a cell-associated form; and (iv) that there was a second, fructose-inducible fructan hydrolase activity produced by these strains. The data provide the first detailed molecular analysis of fructan production and catabolism in this abundant and important oral bacterium. PMID- 11500410 TI - Protective efficacy of a DNA vaccine encoding antigen 85A from Mycobacterium bovis BCG against Buruli ulcer. AB - Buruli ulcer, caused by Mycobacterium ulcerans, is characterized by deep and necrotizing skin lesions, mostly on the arms and legs. Together with tuberculosis and leprosy, this mycobacterial disease has become a major health problem in tropical and subtropical regions, particularly in central and western Africa. No specific vaccine is available for Buruli ulcer. There is, however, evidence in the literature that suggests a cross-reactive protective role of the tuberculosis vaccine M. bovis BCG. To identify potential mechanisms for this cross-protection, we identified and characterized the M. ulcerans homologue of the important protective mycobacterial antigen 85 (Ag85A) from BCG. The homologue is well conserved in M. ulcerans, showing 84.1% amino acid sequence identity and 91% conserved residues compared to the sequence from BCG. This antigen was sufficiently conserved to allow cross-reactive protection, as demonstrated by the ability of M. ulcerans- infected mice to exhibit strong cellular immune responses to both BCG and its purified Ag85 complex. To further address the mechanism of cross-reactive protection, we demonstrate here that prior vaccination with either BCG or plasmid DNA encoding BCG Ag85A is capable of significantly reducing the bacterial load in the footpads of M. ulcerans- infected mice, as determined by Ziehl-Neelsen staining and by actual counting of CFU on 7H11 Middlebrook agar. Together, the results reported here support the potential of a cross-protective Ag85-based future vaccine against tuberculosis, Buruli ulcer, and leprosy. PMID- 11500411 TI - Characterization and protective potential of the immune response to Taenia solium paramyosin in a murine model of cysticercosis. AB - Paramyosin has been proposed as a vaccine candidate in schistosomiasis and filariasis. However, limited information is available about its protective potential against cysticercosis and the immune response it induces. Immunization of mice with recombinant full-length paramyosin of Taenia solium (TPmy) results in about a 52% reduction in parasite burden after a subsequent challenge by intraperitoneal inoculation of Taenia crassiceps cysticerci. Immunization assays using recombinant fragments of TPmy, corresponding approximately to thirds on the amino, central, or carboxyl regions, suggest that protective epitopes are located mostly in the amino-end third. Proliferation assays using T cells obtained from mice immunized with the full-length recombinant TPmy also showed a preferential response to the amino-terminal fragment. In contrast, antibodies in the sera from these mice predominantly recognize epitopes located in the carboxyl-terminal fragment, being the immunoglobulin G1 subclass, the predominant antibody isotype. Characterization of the cellular immune response induced against the protective amino-terminal fragment reveals production of gamma interferon and interleukin-2, but not interleukin-4, suggesting a Th1-like profile. PMID- 11500412 TI - Ontogeny of Th1 memory responses against a Brucella abortus conjugate. AB - Protective immune responses to intracellular pathogens such as Brucella abortus are characteristically Th1-like. Recently we demonstrated that heat-killed B. abortus (HKBa), a strong Th1 stimulus, conjugated to ovalbumin (HKBA-OVA), but not B. abortus alone, can alter the antigen-specific cytokine profile from Th2- to Th1-like. In this report we study the ability of a single injection of B. abortus to switch a Th2 to a Th1 response in immature mice. One-day- and 1-week old mice were given a single injection of B. abortus in the absence or presence of OVA, and at maturity mice were challenged with an allergenic preparation, OVA with alum (OVA-A). B. abortus given without OVA did not diminish the subsequent Th2 response in either age group. In contrast, mice receiving a single injection of B. abortus-OVA at the age of 1 week, but not those injected at the age of 1 day, had reversal of the ratio of OVA-specific Th1 to Th2 cells and decreased immunoglobulin E levels after allergen challenge as adults. Within 6 h both 1-day and 1-week-old mice expressed interleukin-12 p40 mRNA following either B. abortus or B. abortus-OVA administration. However, only the 1-week-old mice exhibited increased expression of gamma interferon (IFN-gamma) mRNA. The absence of the early IFN-gamma response in 1-day-old mice may explain their inability to generate a Th1 memory response. These results suggest that at early stages of immune development, responses to intracellular bacteria may be Th2- rather than Th1-like. Furthermore, they suggest that the first encounter with antigen evokes either a Th1- or a Th2-like response which becomes imprinted, so that subsequent memory responses conform to the original Th bias. This has implications for protection against infectious agents and development of allergic responses. PMID- 11500413 TI - Chlamydia pneumoniae expresses genes required for DNA replication but not cytokinesis during persistent infection of HEp-2 cells. AB - Chlamydia pneumoniae causes community-acquired pneumonia and is associated with several chronic diseases, including asthma and atherosclerosis. The intracellular growth rate of C. pneumoniae slows dramatically during chronic infection, and such persistence leads to attenuated production of new elementary bodies, appearance of morphologically aberrant reticulate bodies, and altered expression of several chlamydial genes. We used an in vitro system to further characterize persistent C. pneumoniae infection, employing both ultrastructural and transcriptional activity measurements. HEp-2 cells were infected with C. pneumoniae (TW-183) at a multiplicity of infection of 3:1, and at 2 h postinfection gamma interferon (IFN-gamma) was added to the medium at 0.15 or 0.50 ng/ml. Treated and untreated cultures were harvested at several times postinfection. RNA was isolated and reverse transcribed, and reverse transcription (RT)-PCR analyses targeting primary transcripts from chlamydial rRNA operons as well as dnaA, polA, mutS, minD, ftsK, and ftsW mRNA were done. Some cultures were fixed and stained for electron microscopic analysis, and a real-time PCR assay was used to assess relative chlamydial chromosome accumulation under each culture condition. The latter assays showed that bacterial chromosome copies accumulated severalfold during IFN-gamma treatment of infected HEp-2 cells, although less accumulation was observed in cells treated with the higher dose. Electron microscopy demonstrated that high-dose IFN-gamma treatment elicited aberrant forms of the bacterium. RT-PCR showed that chlamydial primary rRNA transcripts were present in all IFN-gamma-treated and untreated cell cultures, indicating bacterial metabolic activity. Transcripts from dnaA, polA, mutS, and minD, all of which encode products for bacterial chromosome replication and partition, were expressed in IFN-gamma-treated and untreated cells. In contrast, ftsK and ftsW, encoding products for bacterial cell division, were expressed in untreated cells, but expression was attenuated in cells treated with low-dose IFN-gamma and absent in cells given the high dose of cytokine. Thus, the development of persistence included production of transcripts for DNA replication related, but not cell division-related, genes. These results provide new insight regarding molecular activities that accompany persistence of C. pneumoniae, as well as suggesting requirements for reactivation from persistent to productive growth. PMID- 11500414 TI - Changes in availability of oxygen accentuate differences in capsular polysaccharide expression by phenotypic variants and clinical isolates of Streptococcus pneumoniae. AB - Most isolates of Streptococcus pneumoniae are mixed populations of transparent (T) and opaque (O) colony phenotypes. Differences in the production of capsular polysaccharide (CPS) between O and T variants were accentuated by changes in the environmental concentration of oxygen. O variants demonstrated a 5.2- to 10.6 fold increase in amounts of CPS under anaerobic compared to atmospheric growth conditions, while CPS production remained low under all conditions for T variants. Increased amounts of CPS in O compared to T pneumococci were associated with increased expression of cps-encoded proteins. The inhibitory effect of oxygen on expression of CPS in O variants correlated with decreased tyrosine phosphorylation of CpsD, a tyrosine kinase and regulator of CPS synthesis. Modulation of CpsD expression and its activity by tyrosine phosphorylation may allow the pneumococcus to adapt to the requirements of both colonization, where decreased CPS allows for adherence, and bacteremia, where increased CPS may be required to escape from opsonic clearance. In patients with invasive infection, paired isolates from the same patient were shown to have predominantly a T colony phenotype without phosphotyrosine on CpsD when cultured from the nasopharynx, and an O phenotype that phosphorylates CpsD in response to oxygen when cultured from the blood. Differences in the availability of oxygen, therefore, may be a key factor in allowing for the selection of distinct phenotypes in these two host environments. PMID- 11500415 TI - Production of Neisseria meningitidis transferrin-binding protein B by recombinant Bordetella pertussis. AB - Neisseria meningitidis serogroup B infections are among the major causes of fulminant septicemia and meningitis, especially severe in young children, and no broad vaccine is available yet. Because of poor immunogenicity of the serogroup B capsule, many efforts are now devoted to the identification of protective protein antigens. Among those are PorA and, more recently, transferrin-binding protein B (TbpB). In this study, TbpB of N. meningitidis was genetically fused to the N terminal domain of the Bordetella pertussis filamentous hemagglutinin (FHA), and the fha-tbpB hybrid gene was expressed in B. pertussis either as a plasmid-borne gene or as a single copy inserted into the chromosome. The hybrid protein was efficiently secreted by the recombinant strains, despite its large size, and was recognized by both anti-FHA and anti-TbpB antibodies. A single intranasal administration of recombinant virulent or pertussis-toxin-deficient, attenuated B. pertussis to mice resulted in the production of antigen-specific systemic immunoglobulin G (IgG), as well as local IgG and IgA. The anti-TbpB serum antibodies were of the IgG1, IgG2a, and IgG2b isotypes and were found to express complement-mediated bactericidal activity against N. meningitidis. These observations indicate that recombinant B. pertussis may be a promising vector for the development of a mucosal vaccine against serogroup B meningococci. PMID- 11500416 TI - Cloning, sequencing, and expression of the leukotoxin gene from Fusobacterium necrophorum. AB - Fusobacterium necrophorum is a gram-negative, rod-shaped, anaerobic bacterium that is a primary or secondary etiological agent in a variety of necrotic purulent infections in animals and humans. Included are diseases of cattle such as liver abscesses and foot rot, which have economically important consequences for the cattle industry. The major virulence factor of this bacterium is leukotoxin, a secreted protein of high molecular weight active against leukocytes from ruminants. The screening of a genomic DNA library with polyclonal antisera raised against native affinity-purified leukotoxin and further extension of the sequence using inverse PCR led to the cloning of the entire leukotoxin gene. The leukotoxin gene open reading frame (ORF; lktA) consists of 9,726 bp and encodes a protein of 3,241 amino acids with an overall molecular weight of 335,956. The leukotoxin does not have sequence similarity with any other bacterial leukotoxin. Five truncated overlapping polypeptides covering the whole lktA ORF were used to immunize rabbits. In Western blot assays, polyclonal antisera raised against all five truncated polypeptides recognized affinity-purified leukotoxin from F. necrophorum culture supernatant in a Western blot assay. Antisera directed against two of the five polypeptides had neutralizing activity against the toxin. The entire leukotoxin ORF was expressed in Escherichia coli. Flow-cytometric analysis showed that the recombinant leukotoxin was active against bovine polymorphonuclear leukocytes and was inhibited with antiserum raised against the F. necrophorum leukotoxin. Southern blot hybridization analysis revealed different patterns of lktA hybridizing bands between isolates of the two subspecies of F. necrophorum. PMID- 11500417 TI - Genetic immunization with the region encoding the alpha-helical domain of PspA elicits protective immunity against Streptococcus pneumoniae. AB - Pneumococcal surface protein A (PspA) is a pneumococcal virulence factor capable of eliciting protection against pneumococcal infection in mice. Previous studies have demonstrated that the protection is antibody mediated. Here we examined the ability of pspA to elicit a protective immune response following genetic immunization of mice. Mice were immunized by intramuscular injections with a eukaryotic expression vector encoding the alpha-helical domain of PspA/Rx1. Immunization induced a PspA-specific serum antibody response, and immunized mice survived pneumococcal challenge. Survival and antibody responses occurred in a dose-dependent manner, the highest survival rates being seen with doses of 10 microg or greater. The ability of genetic immunization to elicit cross-protection was demonstrated by the survival of immunized mice challenged with pneumococcal strains differing in capsule and PspA types. Also, immunized mice were protected from intravenous and intratracheal challenges with pneumococci. Similar to the results seen with immunization with PspA, the survival of mice genetically immunized with pspA was antibody mediated. There was no decline in the level of protection 7 months after immunization. These results support the use of genetic immunization to elicit protective immune responses against extracellular pathogens. PMID- 11500418 TI - Purification, characterization, and immunogenicity of a disulfide cross-linked Plasmodium vivax vaccine candidate antigen, merozoite surface protein 1, expressed in Escherichia coli. AB - The Plasmodium vivax merozoite surface protein 1 (MSP-1) 42-kDa fragment (PvMSP-1 p42) is a promising vaccine candidate antigen against the blood stage of the malarial parasite. We have developed a process for the production of this vaccine target, keeping in mind its use in human volunteers. A novel strain, Origami(DE3), of Escherichia coli with mutations in the glutathione and thioredoxin reductase genes yielded 60% more soluble PvMSP-1 p42 than the conventional E. coli BL21(DE3) strain. Recombinant PvMSP-1 p42 was purified to > or = 99% purity with a rapid two-step protocol designed for easy scaling up. The final product had a low endotoxin content and was stable in its lyophilized form. PvMSP-1 p42 was found to have the predicted primary and tertiary structures and consisted of a single conformer containing one free cysteine, as predicted. The product was recognized by conformational monoclonal antibodies against P. vivax MSP-1. Immunogenicity studies of PvMSP-1 p42 were carried out with two strains of mice and the adjuvants Montanide ISA51 and Montanide ISA720. Both formulations were found to induce high levels of immunoglobulin G1 (IgG1), IgG2b, and IgG2a antibodies along with low levels of IgG3. Lymphocytes from animals in all the PvMSP-1 p42-immunized groups showed proliferative responses upon stimulation with PvMSP-1 p42; the cytokines interleukin 2 (IL-2), gamma interferon, IL-4, and IL 10 were detected in the culture supernatants. These results indicate that PvMSP-1 p42 in combination with both of the adjuvants elicited cellular and humoral responses in mice. PMID- 11500419 TI - Salmonella enterica serovar Gallinarum requires the Salmonella pathogenicity island 2 type III secretion system but not the Salmonella pathogenicity island 1 type III secretion system for virulence in chickens. AB - Salmonella enterica serovar Gallinarum is a host-specific serotype that causes the severe systemic disease fowl typhoid in domestic poultry and a narrow range of other avian species but rarely causes disease in mammalian hosts. Specificity of the disease is primarily at the level of the reticuloendothelial system, but few virulence factors have been described other than the requirement for an 85-kb virulence plasmid. In this work, by making functional mutations in the type III secretion systems (TTSS) encoded by Salmonella pathogenicity island 1 (SPI-1) and SPI-2, we investigated the role of these pathogenicity islands in interactions between Salmonella serovar Gallinarum and avian cells in vitro and the role of these pathogenicity islands in virulence in chickens. The SPI-1 mutant showed decreased invasiveness into avian cells in vitro but was unaffected in its ability to persist within chicken macrophages. In contrast the SPI-2 mutant was fully invasive in nonphagocytic cells but failed to persist in macrophages. In chicken infections the SPI-2 mutant was attenuated while the SPI-1 mutant showed full virulence. In oral infections the SPI-2 mutant was not observed in the spleen or liver, and following intravenous inoculation it was cleared rapidly from these sites. SPI-2 function is required by Salmonella serovar Gallinarum for virulence, primarily through promoting survival within macrophages allowing multiplication within the reticuloendothelial system, but this does not preclude the involvement of SPI-2 in uptake from the gut to the spleen and liver. SPI-1 appears to have little effect on virulence and survival of Salmonella serovar Gallinarum in the host. PMID- 11500420 TI - DNA sequences encoding CD4+ and CD8+ T-cell epitopes are important for efficient protective immunity induced by DNA vaccination with a Trypanosoma cruzi gene. AB - Immunization of BALB/c mice with a plasmid containing the gene for Trypanosoma cruzi trans-sialidase (TS) induced antibodies that inhibited TS enzymatic activity, CD4+ Th1 and CD8+ Tc1 cells, and protective immunity against infection. We used this model to obtain basic information on the requirement of CD4 or CD8 or B-cell epitopes for an effective DNA-induced immunity against T. cruzi infection. For that purpose, mice were immunized with plasmids containing DNA sequences encoding (i) the entire TS protein, (ii) the TS enzymatic domain, (iii) the TS CD4+ T-cell epitopes, (iv) the TS CD8+ T-cell epitope, or (v) TS CD4+ and CD8+ T-cell epitopes. Plasmids expressing the entire TS or its enzymatic domain elicited similar levels of TS-inhibitory antibodies, gamma interferon (IFN-gamma) producing T cells, and protective immunity against infection. Although the plasmid expressing TS CD4 epitopes was immunogenic, its protective efficacy against experimental infection was limited. The plasmid expressing the CD8 epitope was poorly immunogenic and provided little protective immunity. The reason for the limited priming of CD8+ T cells was due to a requirement for CD4+ T cells. To circumvent this problem, a plasmid expressing both CD4+ and CD8+ T cell epitopes was produced. This plasmid generated levels of IFN-gamma-producing T cells and protective immunity comparable to that of the plasmid expressing the entire catalytic domain of TS. Our observations suggest that plasmids expressing epitopes recognized by CD4+ and CD8+ T cells may have a better protective potential against infection with T. cruzi. PMID- 11500421 TI - pH-enhanced cytopathic effects of Clostridium sordellii lethal toxin. AB - Clostridium sordellii lethal toxin (TcsL) is a large clostridial toxin (LCT) that glucosylates Ras, Rac, and Ral. TcsL differs from other LCTs because it modifies Ras, which does not cycle from cytosol to membrane. By using a suite of inhibitors, steps in cell entry by TcsL were dissected, and entry appears to be dependent on endosomal acidification. However, in contrast to TcdB, TcsL was substantially slower in its time course of entry. TcsL cytopathic effects (CPE) were blocked by bafilomycin A1 and neutralized by antiserum up to 2 h following treatment of cells with the toxin. The slow time course of intoxication and relatively high cytopathic dose were alleviated by exposing TcsL to acid pH, resulting in a time course similar to that of TcdB. The optimal pH range for activation was 4.0 to 5.0, which increased the rate of intoxication over 5-fold, lowered the minimal intoxicating dose by over 100-fold, and allowed complete substrate modification within 2 h, as shown by differential glucosylation. Fluorescence analysis of TcsL with 2-(p-toluidinyl) naphthalene-6-sulfonic acid as a probe suggested the acid pH stimulated a hydrophobic transition in the protein, a likely prelude to membrane insertion. Finally, acid entry by TcsL caused TcdB-like morphological changes in CHO cells, which suggesting that acid activation may impact substrate recognition profiles for TcsL. PMID- 11500422 TI - Novel extracellular x-prolyl dipeptidyl-peptidase (DPP) from Streptococcus gordonii FSS2: an emerging subfamily of viridans Streptococcal x-prolyl DPPs. AB - Streptococcus gordonii is generally considered a benign inhabitant of the oral microflora, and yet it is a primary etiological agent in the development of subacute bacterial endocarditis (SBE), an inflammatory state that propagates thrombus formation and tissue damage on the surface of heart valves. Strain FSS2 produced several extracellular aminopeptidase and fibrinogen-degrading activities during growth in culture. In this report we describe the purification, characterization, and cloning of a serine class dipeptidyl-aminopeptidase, an x prolyl dipeptidyl-peptidase (Sg-xPDPP, for S. gordonii x-prolyl dipeptidyl peptidase), produced in a pH-controlled batch culture. Purification of this enzyme by anion exchange, gel filtration, and hydrophobic interaction chromatography yielded a protein monomer of approximately 85 kDa, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) under denaturing conditions. However, under native conditions, the protein appeared to be a homodimer on the basis of gel filtration and PAGE. Kinetic studies indicated that purified enzyme had a unique and stringent x-prolyl specificity that is comparable to both the dipeptidyl-peptidase IV/CD26 and lactococcal x-prolyl dipeptidyl-peptidase families. Nested PCR cloning from an S. gordonii library enabled the isolation and sequence analysis of the full-length gene. A 759-amino acid polypeptide with a theoretical molecular mass of 87,115 Da and a calculated pI of 5.6 was encoded by this open reading frame. Significant homology was found with the PepX gene family from Lactobacillus and Lactococcus spp. and putative x prolyl dipeptidyl-peptidases from other streptococcal species. Sg-xPDPP may serve as a critical factor for the sustained bacterial growth in vivo and furthermore may aid in the proteolysis of host tissue that is commonly observed during SBE pathology. PMID- 11500423 TI - Induction of specific cytotoxic lymphocytes in mice vaccinated with Brucella abortus RB51. AB - A safe, more sensitive, nonradioactive, neutral red uptake assay was adopted to replace the traditional 51Cr release assay for detection of Brucella-specific cytotoxic T lymphocyte (CTL) activity. Our studies indicated that Brucella abortus strain RB51 vaccination of mice induced specific CTLs against both strain RB51- and strain 2308-infected J774.A1 macrophages but not against Listeria monocytogenes-infected J774.A1 cells. The antigen-specific cytotoxic activity was exerted by T lymphocytes but not by NK cells. CD3+ CD4+ T cells secreted the highest level of gamma interferon (IFN-gamma) and were able to exert a low but significant level of specific lysis of Brucella-infected macrophages. They also exerted a low level of nonspecific lysis of noninfected macrophages. In contrast, CD3+ CD8+ T cells secreted low levels of IFN-gamma but demonstrated high levels of specific lysis of Brucella-infected macrophages with no nonspecific lysis. These findings indicate that B. abortus strain RB51 vaccination of mice induces specific CTLs and suggest that CD3+ CD4+ and CD3+ CD8+ T cells play a synergistic role in the anti-Brucella activity. PMID- 11500424 TI - Neisseria meningitidis RTX protein FrpC induces high levels of serum antibodies during invasive disease: polymorphism of frpC alleles and purification of recombinant FrpC. AB - The Neisseria meningitidis FAM20 strain secretes two proteins of unknown function, FrpA and FrpC, which contain typical RTX domains found in cytotoxins from other gram-negative pathogens. To evaluate whether the Frp proteins could be involved in meningococcal virulence, 65 isolates of all serogroups were screened by PCR for the presence of both frp genes. The frpA allele was, however, poorly conserved. A single strain harbored an frpA allele of the previously described size, while large insertions were detected in the frpA loci of 22 isolates (34%), and the 42 remaining isolates (65%) did not contain frpA at all. In contrast, frpC alleles, albeit of variable length, were detected in all invasive and most carrier strains. This suggests that meningococci may produce a family of FrpC proteins of various molecular masses. High levels of both immunoglobulin G (IgG) and IgA class antibodies recognizing recombinant FrpC were indeed detected in convalescent-phase sera of most patients at 2 and 4 to 5 weeks after the first symptoms of meningococcal disease. These results show that FrpC-like proteins are produced and may play a role in invasive meningococcal infections. PMID- 11500425 TI - Genotypic variation in the Bordetella pertussis virulence factors pertactin and pertussis toxin in historical and recent clinical isolates in the United Kingdom. AB - The reemergence of pertussis has been reported in several countries despite high vaccination coverage. Studies in The Netherlands and Finland have investigated polymorphism in the genes coding for two important virulence factors of Bordetella pertussis, pertactin and pertussis toxin, and identified the emergence and subsequent dominance in circulating strains of pertactin and toxin variants not found in the whole-cell vaccine (WCV). The study described here investigated whether such variation had occurred in the United Kingdom, which presently has low levels of pertussis. Sequence analysis of the genes for pertactin (prnA) and the pertussis toxin S1 subunit (ptxA) among isolates of B. pertussis from 285 United Kingdom patients, from 1920 to 1999, revealed three prnA variants, prnA(1), prnA(2), and prnA(3), and two ptxA variants, ptxA(1) and ptxA(2), showing differences in nucleic acid sequence. The proportion of pertactin gene types not included in the United Kingdom WCV, i.e., prnA(2) and prnA(3), has increased in recent years and was found in 21 of 86 (24%) strains from the 1980s and 56 of 105 (53%) strains from the 1990s. To date, the presence of these nonvaccine prnA types has not been associated with a resurgence of pertussis in the United Kingdom. The distribution of prnA and ptxA types in The Netherlands, Finland, and the United Kingdom in the 1990s is distinct. The most striking difference in the United Kingdom isolates is that all 105 of the most recent circulating strains (from 1998 to 1999) are of a pertussis toxin type found in the United Kingdom WCV, i.e., ptxA(1). PMID- 11500426 TI - Adherent invasive Escherichia coli strains from patients with Crohn's disease survive and replicate within macrophages without inducing host cell death. AB - Escherichia coli strains recovered from Crohn's disease (CD) lesions are able to adhere to and invade cultured intestinal epithelial cells. We analyzed the behavior within macrophages of adherent invasive E. coli (AIEC) strains isolated from patients with CD. All the 15 AIEC strains tested were able to replicate extensively within J774-A1 cells: the numbers of intracellular bacteria increased 2.2- to 74.2-fold at 48 h over that at 1 h postinfection. By use of murine peritoneal macrophages and human monocyte-derived-macrophages, the reference AIEC strain LF82 was confirmed to be able to survive intracellularly. Transmission electron micrographs of AIEC LF82-infected macrophages showed that at 24 h postinfection, infected cells harbored large vacuoles containing numerous bacteria, as a result of the fusion of several vacuoles occurring after 8 h postinfection. No lactate dehydrogenase (LDH) release, no sign of DNA fragmentation or degradation, and no binding to fluorescein isothlocyanate labeled annexin V were observed with LF82-infected J774-A1 cells, even after 24 h postinfection. LF82-infected J774-A1 cells secreted 2.7-fold more tumor necrosis factor alpha (TNF-alpha) than cells stimulated with 1 microg of lipopolysaccharide (LPS)/ml. No release of interleukin-1beta was observed with LPS-prestimulated J774-A1 cells infected with AIEC LF82. These findings showed that (i) AIEC strains are able to survive and to replicate within macrophages, (ii) AIEC LF82 replication does not induce any cell death of the infected cells, and (iii) LF82-infected J774-A1 cells release high levels of TNF-alpha. These properties could be related to some features of CD and particularly to granuloma formation, one of the hallmarks of CD lesions. PMID- 11500427 TI - Conserved DegP protease in gram-positive bacteria is essential for thermal and oxidative tolerance and full virulence in Streptococcus pyogenes. AB - The DegP protease, a multifunctional chaperone and protease, has been shown to be essential for virulence in gram-negative pathogens such as Salmonella enterica serovar Typhimurium, Brucella abortus, Yersinia enterocolitica, and Pseudomonas aeruginosa. The function of DegP in pathogenesis appears to be the degradation of damaged proteins that accumulate as a result of the initial host response to infection, which includes the release of reactive oxygen intermediates. Additionally, the DegP protease plays a major role in monitoring and maintaining the Escherichia coli periplasm and influences E. coli pilus biogenesis. We report here the identification of highly homologous enzymes in Streptococcus pyogenes, Streptococcus gordonii, Streptococcus mutans, Staphylococcus aureus, and Enterococcus faecalis. Moreover, the phenotype of an insertionally inactivated degP allele in S. pyogenes is similar to that reported for E. coli, with temperature sensitivity for growth and enhanced sensitivity to reactive oxygen intermediates. Virulence studies in a mouse model of streptococcal infection indicate that a functional DegP protease is required for full virulence. These results suggest DegP as an attractive broad-spectrum target for future anti infective drug development. PMID- 11500428 TI - Role of the heat shock protein 90 in immune response stimulation by bacterial DNA and synthetic oligonucleotides. AB - To elucidate the mechanisms of immunostimulation by bacterial DNA and synthetic oligonucleotides, the effects of heat shock protein 90 (Hsp90) inhibitors on the activation of murine spleen cells and macrophages by these molecules were investigated. Murine spleen cells and J774 and RAW264.7 macrophages responded to a CpG-containing oligodeoxynucleotide (CpG ODN) and Escherichia coli DNA by increased production of interleukin 6 (IL-6), IL-12, tumor necrosis factor alpha, and nitric oxide (NO). Pretreatment with any of the three Hsp90 inhibitors geldanamycin, radicicol, and herbimycin A resulted in a dose-dependent suppression of cytokine production from the spleen cells and macrophages and of NO from macrophages stimulated with CpG ODN or E. coli DNA. These Hsp90 inhibitors, however, had no effect on Staphylococcus aureus Cowan strain 1 induced IL-12 production from either the murine spleen cells or macrophages. CpG ODN and E. coli DNA induced increased intracellular levels of phosphorylated extracellular signal-regulated kinases (ERK1 and -2), which are members of the mitogen-activated protein (MAP) kinase family, while geldanamycin and radicicol blocked the phosphorylation of ERK1 and -2 in J774 and RAW264.7 cells. These data indicate that DNA-induced activation of murine spleen cells and macrophages is mediated by Hsp90 and that Hsp90 inhibitor suppression of DNA-induced macrophage activation is associated with disruption of the MAP kinase signaling pathway. Our findings suggest that Hsp90 inhibitors may provide a useful means of elucidating the mechanisms of immunostimulation by bacterial DNA and CpG ODN as well as a strategy for preventing adverse effects of bacterial DNA as well as lipopolysaccharide. PMID- 11500429 TI - Comparative sequence analysis of the plasmid-encoded regulator of enteropathogenic Escherichia coli Strains. AB - Enteropathogenic Escherichia coli (EPEC) strains that carry the EPEC adherence factor (EAF) plasmid were screened for the presence of different EAF sequences, including those of the plasmid-encoded regulator (per). Considerable variation in gene content of EAF plasmids from different strains was seen. However, bfpA, the gene encoding the structural subunit for the bundle-forming pilus, bundlin, and per genes were found in 96.8% of strains. Sequence analysis of the per operon and its promoter region from 15 representative strains revealed that it is highly conserved. Most of the variation occurs in the 5' two-thirds of the perA gene. In contrast, the C-terminal portion of the predicted PerA protein that contains the DNA-binding helix-turn-helix motif is 100% conserved in all strains that possess a full-length gene. In a minority of strains including the O119:H2 and canine isolates and in a subset of O128:H2 and O142:H6 strains, frameshift mutations in perA leading to premature truncation and consequent inactivation of the gene were identified. Cloned perA, -B, and -C genes from these strains, unlike those from strains with a functional operon, failed to activate the LEE1 operon and bfpA transcriptional fusions or to complement a per mutant in reference strain E2348/69. Furthermore, O119, O128, and canine strains that carry inactive per operons were deficient in virulence protein expression. The context in which the perABC operon occurs on the EAF plasmid varies. The sequence upstream of the per promoter region in EPEC reference strains E2348/69 and B171-8 was present in strains belonging to most serogroups. In a subset of O119:H2, O128:H2, and O142:H6 strains and in the canine isolate, this sequence was replaced by an IS1294-homologous sequence. PMID- 11500430 TI - Multistage multiantigen heterologous prime boost vaccine for Plasmodium knowlesi malaria provides partial protection in rhesus macaques. AB - A nonhuman primate model for malaria vaccine development allowing reliable, stringent sporozoite challenge and evaluation of both cellular and antibody responses is needed. We therefore constructed a multicomponent, multistage DNA vaccine for the simian malaria species Plasmodium knowlesi including two preerythrocytic-stage antigens, the circumsporozoite protein (PkCSP) and sporozoite surface protein 2 (PkSSP2), and two blood stage antigens, apical merozoite antigen 1 (PkAMA1) and merozoite surface protein 1 (PkMSP1p42), as well as recombinant canarypox viruses encoding the four antigens (ALVAC-4). The DNA vaccine plasmids expressed the corresponding antigens in vitro and induced antiparasite antibodies in mice. Groups of four rhesus monkeys received three doses of a mixture of the four DNA vaccine plasmids and a plasmid encoding rhesus granulocyte-monocyte colony-stimulating factor, followed by boosting with a single dose of ALVAC-4. Three groups received the priming DNA doses by different routes, either by intramuscular needle injection, by intramuscular injection with a needleless injection device, the Biojector, or by a combination of intramuscular and intradermal routes by Biojector. Animals immunized by any route developed antibody responses against sporozoites and infected erythrocytes and against a recombinant PkCSP protein, as well as gamma interferon-secreting T-cell responses against peptides from PkCSP. Following challenge with 100 P. knowlesi sporozoites, 1 of 12 experimental monkeys was completely protected and the mean parasitemia in the remaining monkeys was significantly lower than that in 4 control monkeys. This model will be important in preclinical vaccine development. PMID- 11500431 TI - Gamma interferon-induced inhibition of Toxoplasma gondii in astrocytes is mediated by IGTP. AB - Toxoplasma gondii is an important pathogen in the central nervous system, causing a severe and often fatal encephalitis in patients with AIDS. Gamma interferon (IFN-gamma) is the main cytokine preventing reactivation of Toxoplasma encephalitis in the brain. Microglia are important IFN-gamma-activated effector cells controlling the growth of T. gondii in the brain via a nitric oxide (NO) mediated mechanism. IFN-gamma can also activate astrocytes to inhibit the growth of T. gondii. Previous studies found that the mechanism in murine astrocytes is independent of NO and all other known anti-Toxoplasma mechanisms. In this study we investigated the role of IGTP, a recently identified IFN-gamma-regulated gene, in IFN-gamma inhibition of T. gondii in murine astrocytes. Primary astrocytes were cultivated from IGTP-deficient mice, treated with IFN-gamma, and then tested for anti-Toxoplasma activity. In wild-type astrocytes T. gondii growth was significantly inhibited by IFN-gamma, whereas in astrocytes from IGTP-deficient mice IFN-gamma did not cause a significant inhibition of growth. Immunoblot analysis confirmed that IFN-gamma induced significant levels of IGTP in wild-type murine astrocytes within 24 h. These results indicate that IGTP plays a central role in the IFN-gamma-induced inhibition of T. gondii in murine astrocytes. PMID- 11500432 TI - Exploitation of interleukin-8-induced neutrophil chemotaxis by the agent of human granulocytic ehrlichiosis. AB - The agent of human granulocytic ehrlichiosis (HGE) is an obligate intracellular bacterium with a tropism for neutrophils; however, the mechanisms of bacterial dissemination are not yet understood. Interleukin-8 (IL-8) is a chemokine that induces neutrophil migration to sites of infection for host defense against pathogens. We now show that HGE bacteria, and the HGE-44 protein, induce IL-8 secretion in a promyelocytic (HL-60) cell line that has been differentiated along the neutrophil lineage with retinoic acid and in neutrophils. Infected HL-60 cells also demonstrate upregulation of CXCR2, an IL-8 receptor, but not CXCR1. Human neutrophils migrate towards Ehrlichia sp.-infected cells in a chemotaxis chamber assay, and this movement can be blocked with antibodies to IL-8. Finally, immunocompetent and severe combined immunodeficient mice administered CXCR2 antisera, and CXCR2(-/-) mice that lack the human IL-8 receptor homologue, are much less susceptible to granulocytic ehrlichiosis than are control animals. These results demonstrate that HGE bacteria induce IL-8 production by host cells and, paradoxically, appear to exploit this chemokine to enhance infection. PMID- 11500433 TI - Laccase of Cryptococcus neoformans is a cell wall-associated virulence factor. AB - Virulence is the outcome of an interaction between the host and a microbe and is characterized by a large array of opposing reactions operating at the host pathogen interface. Cryptococcus neoformans is an important opportunistic pathogen in immunocompromised patients, including those with human immunodeficiency virus, and expresses a virulence-associated laccase which is believed to oxidize brain catecholamines and iron as a defense against host immune cells. In the present report, we investigated the cellular location of laccase to understand more fully how it contributes to cryptococcal virulence. A monoclonal antibody to the C. neoformans laccase was generated and used to show localization in the cell walls of representative serotype A (H99) and serotype D (B-3501) strains by immunoelectron microscopy. In addition, confocal microscopy was used to show a peripheral location of green fluorescent protein-tagged laccase expressed in live H99 cells. Biochemical studies showed that laccase could be released from intact cells or cell wall fractions with glucanase enzymes but was retained in the cell wall after sequential extraction with 1 M NaCl, 6 M urea, and 1% sodium dodecyl sulfate. The presence of a hydrolyzable bond linking laccase to the cell wall was suggested by removal of laccase from cell wall preparations after they were boiled in 1% sodium dodecyl sulfate, as was the presence of a disulfide or thioester bond by removal with dithiothreitol or beta mercaptoethanol. These data show that laccase is present as a tightly associated cell wall enzyme that is readily accessible for interactions with host immune cells. PMID- 11500434 TI - Intimin-specific immune responses prevent bacterial colonization by the attaching effacing pathogen Citrobacter rodentium. AB - The formation of attaching and effacing (A/E) lesions on gut enterocytes is central to the pathogenesis of enterohemorrhagic (EHEC) Escherichia coli, enteropathogenic E. coli (EPEC), and the rodent pathogen Citrobacter rodentium. Genes encoding A/E lesion formation map to a chromosomal pathogenicity island termed the locus of enterocyte effacement (LEE). Here we show that the LEE encoded proteins EspA, EspB, Tir, and intimin are the targets of long-lived humoral immune responses in C. rodentium-infected mice. Mice infected with C. rodentium developed robust acquired immunity and were resistant to reinfection with wild-type C. rodentium or a C. rodentium derivative, DBS255(pCVD438), which expressed intimin derived from EPEC strain E2348/69. The receptor-binding domain of intimin polypeptides is located within the carboxy-terminal 280 amino acids (Int280). Mucosal and systemic vaccination regimens using enterotoxin-based adjuvants were employed to elicit immune responses to recombinant Int280alpha from EPEC strain E2348/69. Mice vaccinated subcutaneously with Int280alpha, in the absence of adjuvant, were significantly more resistant to oral challenge with DBS255(pCVD438) but not with wild-type C. rodentium. This type-specific immunity could not be overcome by employing an exposed, highly conserved domain of intimin (Int388-667) as a vaccine. These results show that anti-intimin immune responses can modulate the outcome of a C. rodentium infection and support the use of intimin as a component of a type-specific EPEC or EHEC vaccine. PMID- 11500435 TI - Comparative immune response to PE and PE_PGRS antigens of Mycobacterium tuberculosis. AB - Sequencing of the entire genome of Mycobacterium tuberculosis identified a novel multigene family composed of two closely related subfamilies designated PE and PE_PGRS. The major difference between these two families is the presence of a domain containing numerous Gly-Ala repeats extending to the C terminus of the PE_PGRS genes. We have used a representative PE_PGRS gene from M. tuberculosis, Rv1818c (1818PE_PGRS), and its amino-terminal PE region (1818PE), to investigate the immunological response to these proteins during experimental tuberculosis and following immunization with DNA constructs. During infection of mice with M. tuberculosis, a significant humoral immune response was observed against recombinant 1818PE_PGRS but not toward the 1818PE protein. Similarly, immunization with a 1818PE_PGRS DNA construct induced antibodies directed against 1818PE_PGRS but not against 1818PE proteins, and no humoral response was induced by 1818PE DNA. These results suggest that certain PE_PGRS genes are expressed during infection of the host with M. tuberculosis and that an antibody response is directed solely against the Gly-Ala-rich PGRS domain. Conversely, splenocytes from 1818PE-vaccinated mice but not mice immunized with 1818PE_PGRS secreted gamma interferon following in vitro restimulation and demonstrated protection in the mouse tuberculosis challenge model. These results suggest that the PE vaccine can elicit an effective cellular immune response and that immune recognition of the PE antigen is influenced by the Gly-Ala-rich PGRS domain. PMID- 11500436 TI - Mycoplasma pneumoniae P1 type 1- and type 2-specific sequences within the P1 cytadhesin gene of individual strains. AB - Mycoplasma pneumoniae strains traditionally are divided into two types, based on sequence variation in the P1 gene. Recently, however, we have identified 8 P1 subtypes by restriction fragment length polymorphism analysis. In the present study the P1 gene sequences of three P1 type 1 and two P1 type 2 M. pneumoniae strains were analyzed. A new P1 gene sequence in a type 1 strain with partial similarity to a recently reported variable region in the P1 gene of an M. pneumoniae type 2 strain (T. Kenri, R. Taniguchi, Y. Sasaki, N. Okazaki, M. Narita, K. Izumikawa, M. Umetsu, and T.Sasaki, Infect. Immun. 67:4557-4562, 1999) was identified. In addition, the P1 gene of the type 1 strain contained another region with nucleotide polymorphisms identical to a stretch in the P1 gene of one of our type 2 strains. These findings indicate that recombination between sequences specific for P1 type 1 and type 2 had occurred and that P1 type 1 and type 2 hybrid sequences can be present within the P1 gene of an individual strain. Identical or nearly identical variable P1 gene sequences were present in several repetitive regions outside the P1 gene locus in the genome of M. pneumoniae strain M129, implying recombination as a mechanism for generation of the P1 gene variation. Additionally, in the P1 gene sequences of four of the five strains studied, single-nucleotide polymorphisms different from the previously reported P1 type 1 and 2 characteristic sequences were identified. The polymorphic sites are candidate targets for genotyping of M. pneumoniae by direct sequencing of amplicons from clinical specimens. PMID- 11500437 TI - Absence of all components of the flagellar export and synthesis machinery differentially alters virulence of Salmonella enterica serovar Typhimurium in models of typhoid fever, survival in macrophages, tissue culture invasiveness, and calf enterocolitis. AB - In this study, we constructed an flhD (the master flagellar regulator gene) mutant of Salmonella enterica serovar Typhimurium and compared the virulence of the strain to that of the wild-type strain in a series of assays that included the mouse model of typhoid fever, the mouse macrophage survival assay, an intestinal epithelial cell adherence and invasion assay, and the calf model of enterocolitis. We found that the flhD mutant was more virulent than its parent in the mouse and displayed slightly faster net growth between 4 and 24 h of infection in mouse macrophages. Conversely, the flhD mutant exhibited diminished invasiveness for human and mouse intestinal epithelial cells, as well as a reduced capacity to induce fluid secretion and evoke a polymorphonuclear leukocyte response in the calf ligated-loop assay. These findings, taken with the results from virulence assessment assays done on an fljB fliC mutant of serovar Typhimurium that does not produce flagellin but does synthesize the flagellar secretory apparatus, indicate that neither the presence of flagella (as previously reported) nor the synthesis of the flagellar export machinery are necessary for pathogenicity of the organism in the mouse. Conversely, the presence of flagella is required for the full invasive potential of the bacterium in tissue culture and for the influx of polymorphonuclear leukocytes in the calf intestine, while the flagellar secretory components are also necessary for the induction of maximum fluid secretion in that enterocolitis model. A corollary to this conclusion is that, as has previously been surmised but not demonstrated in a comparative investigation of the same mutant strains, the mouse systemic infection and macrophage assays measure aspects of virulence different from those of the tissue culture invasion assay, and the latter is more predictive of findings in the calf enterocolitis model. PMID- 11500439 TI - Association of interleukin-10 cytokine expression status with HLA non-DRB1*02 and Mycobacterium bovis BCG scar-negative status in south Indian pulmonary tuberculosis patients. AB - HLA DRB1*02 and its subtypes predispose individuals for a far-advanced sputum positive pulmonary tuberculosis transcending ethnic boundaries. Mycobacterium bovis BCG does not afford the desired protection against adult pulmonary tuberculosis, and a spectrum of immune reactivity exists in controls and hospital contacts. All of these findings have been identified and demonstrated in areas of endemicity. Skewing of immunity from protective to pathogenic may involve a shift in the Th1-Th2 paradigm. To elaborate these ideas, we studied gamma interferon (IFN-gamma), interleukin-4 (IL-4), and IL-10 cytokine expression in 71 adult pulmonary tuberculosis patients and 74 controls from areas of endemicity in south India by 48-h microculture and reverse transcription-PCR. Most of the patients and controls expressed IFN-gamma de novo, and in the presence of purified protein derivative (PPD), all of them expressed significantly higher levels of IFN-gamma, suggesting a PPD-specific recall memory. HLA DRB1* allele-dependent IFN-gamma expression was identified only in controls, suggesting a skewing of the immune response in patients. In contrast to the case for IFN-gamma, only some patients and controls expressed IL-4 or IL-10 (Th2 profile); thus, the Th1 profile was identifiable only by a nonexpression of IL-4 or IL-10 in this area of endemicity. The Th2 profile was associated with HLA non-DRB1*02 and BCG scar-negative status in patients, attributing a significant risk (odds ratio = 2.074; 95% confidence interval = 0.612 to 7.07). It is possible that Mycobacterium tuberculosis (PPD) specific IL-10 is expressed preemptively in unvaccinated (BCG scar-negative) individuals with a non-DR2 genetic background by chronic exposure in this area of endemicity and leads to pulmonary tuberculosis of adults. PMID- 11500438 TI - Characterization of Haemophilus ducreyi cdtA, cdtB, and cdtC mutants in in vitro and in vivo systems. AB - Haemophilus ducreyi expresses a soluble cytolethal distending toxin (CDT) that is encoded by the cdtABC gene cluster and can be detected in culture supernatant fluid by its ability to kill HeLa cells. The cdtA, cdtB, and cdtC genes of H. ducreyi were cloned independently into plasmid vectors, and their encoded proteins expressed singly or in various combinations in an Escherichia coli background. All three gene products had to be expressed in order for E. coli derived culture supernatant fluids to demonstrate cytotoxicity for HeLa cells. Isogenic H. ducreyi cdtA and cdtB mutants were constructed and used in combination with the wild-type parent strain and a previously described H. ducreyi cdtC mutant (M. K. Stevens, J. L. Latimer, S. R. Lumbley, C. K. Ward, L. D. Cope, T. Lagergard, and E. J. Hansen, Infect. Immun. 67:3900-3908, 1999) to determine the relative contributions of the CdtA, CdtB, and CdtC proteins to CDT activity. Expression of CdtA, CdtB, and CdtC appeared necessary for H. ducreyi derived culture supernatant fluid to exhibit cytotoxicity for HeLa cells. Whole cell sonicates and periplasmic extracts from the cdtB and cdtC mutants had no effect on HeLa cells, whereas these same fractions from a cdtA mutant had a very modest cytotoxic effect on these same human cells. CdtA appeared to be primarily associated with the H. ducreyi cell envelope, whereas both CdtB and CdtC were present primarily in the soluble fraction from sonicated cells. Both the cdtA mutant and the cdtB mutant were found to be fully virulent in the temperature dependent rabbit model for experimental chancroid. PMID- 11500440 TI - Cardiac myosin autoimmunity in acute Chagas' heart disease. AB - Infection with Trypanosoma cruzi, the agent of Chagas' disease, may induce antibodies and T cells reactive with self antigens (autoimmunity). Because autoimmunity is generally thought to develop during the chronic phase of infection, one hypothesis is that autoimmunity develops only after long-term, low level stimulation of self-reactive cells. However, preliminary reports suggest that autoimmunity may begin during acute T. cruzi infection. The goal of the present study was to investigate whether cardiac autoimmunity could be observed during acute T. cruzi infection. A/J mice infected with the Brazil strain of T. cruzi for 21 days developed severe myocarditis, accompanied by humoral and cellular autoimmunity. Specifically, T. cruzi infection induced immunoglobulin G (IgG) autoantibodies and delayed type hypersensitivity (DTH) to cardiac myosin. This autoimmunity resembles that which develops in A/J mice immunized with myosin in complete Freund's adjuvant in that myosin-specific antibodies and DTH responses both develop by 21 days postinfection or postimmunization. While the levels of myosin IgG in T. cruzi-infected mice were slightly lower than those in myosin-immunized mice, the magnitude of myosin DTH in the two groups was statistically equivalent. In contrast, C57BL/6 mice, which are resistant to myosin-induced myocarditis and its associated autoimmunity, developed undetectable or low levels of myosin IgG and did not exhibit myosin DTH or myocarditis upon T. cruzi infection. Therefore, humoral and cellular cardiac autoimmunity can develop during acute T. cruzi infection in the genetically susceptible host. PMID- 11500441 TI - Hydrolysis of interleukin-12 by Porphyromonas gingivalis major cysteine proteinases may affect local gamma interferon accumulation and the Th1 or Th2 T cell phenotype in periodontitis. AB - Porphyromonas gingivalis cysteine proteinases (gingipains) have been associated with virulence in destructive periodontitis, a disease process variously considered to represent an unregulated stimulation of either T helper type 1 (Th1)- or Th2-type cells. Critical in maintaining Th1 activity is the response of T lymphocytes to environmental interleukin 12 (IL-12) in the form of up regulation of gamma interferon (IFN-gamma) production. Here we demonstrate that in the presence or absence of serum, gingipains were able to hydrolyze IL-12 and reduce the IL-12-induced IFN-gamma production from CD4+ T cells. However, the induction of IL-12 receptors on T cells by gingipains did not correlate with the enhancement of IFN-gamma production. The gingipains cleaved IL-12 within the COOH terminal region of the p40 and p35 subunit chains, which leads to IL-12 inactivity, whereas IL-2 in these assays was not affected. Inactivation of IL-12 by the gingipains could disrupt the cytokine balance or favor Th2 activities in the progression of periodontitis. PMID- 11500442 TI - Production of matrix metalloproteinases in response to mycobacterial infection. AB - Matrix metalloproteinases (MMPs) constitute a large family of enzymes with specificity for the various proteins of the extracellular matrix which are implicated in tissue remodeling processes and chronic inflammatory conditions. To investigate the role of MMPs in immunity to mycobacterial infections, we incubated murine peritoneal macrophages with viable Mycobacterium bovis BCG or Mycobacterium tuberculosis H37Rv and assayed MMP activity in the supernatants by zymography. Resting macrophages secreted only small amounts of MMP-9 (gelatinase B), but secretion increased dramatically in a dose-dependent manner in response to either BCG or M. tuberculosis in vitro. Incubation with mycobacteria also induced increased MMP-2 (gelatinase A) activity. Neutralization of tumor necrosis alpha (TNF-alpha), and to a lesser extent interleukin 18 (IL-18), substantially reduced MMP production in response to mycobacteria. Exogenous addition of TNF alpha or IL-18 induced macrophages to express MMPs, even in the absence of bacteria. The immunoregulatory cytokines gamma interferon (IFN-gamma), IL-4, and IL-10 all suppressed BCG-induced MMP production, but through different mechanisms. IFN-gamma treatment increased macrophage secretion of TNF-alpha but still reduced their MMP activity. Conversely, IL-4 and IL-10 seemed to act by reducing the amount of TNF-alpha available to the macrophages. Finally, infection of BALB/c or severe combined immunodeficiency (SCID) mice with either BCG or M. tuberculosis induced substantial increases in MMP-9 activity in infected tissues. In conclusion, we show that mycobacterial infection induces MMP-9 activity both in vitro and in vivo and that this is regulated by TNF-alpha, IL-18, and IFN gamma. These findings indicate a possible contribution of MMPs to tissue remodeling processes that occur in mycobacterial infections. PMID- 11500443 TI - Expression of a Mycobacterium tuberculosis arabinomannan antigen in vitro and in vivo. AB - The outermost layer of Mycobacterium tuberculosis contains two major polysaccharides, arabinomannan (AM) and glucan (GC). We studied the in vitro and in vivo expression of an M. tuberculosis AM antigen using monoclonal antibody (MAb) 9d8 (2a), an isotype-switched variant of the immunoglobulin G3 (IgG3) MAb 9d8. MAb 9d8 had been previously shown to bind M. tuberculosis AM and the M. tuberculosis surface. Our in vitro experiments showed that MAb 9d8(2a) bound strongly to whole-cell M. tuberculosis Erdman but not to the CDC 1551 strain grown in medium for an extended period. However, AM antigen was detected in the culture supernatant of both strains, and its concentration increased in a time dependent manner. The detection of AM antigen from both strains was decreased in the presence of Tween 80. In mice infected with M. tuberculosis Erdman, AM antigen accumulated in organ homogenates concomitant to an increase in bacterial organ burden and an increase in IgG and IgM titer to AM. These results (i) indicate that the surface expression of AM during in vitro growth changes with culture age, is strain dependent, and is affected by the presence of Tween 80 in the culture media; (ii) show that AM is produced by bacteria growth in vivo; and (iii) demonstrate that the amount of in vivo-detected AM can be dependent on the number of bacteria in the infected organ. PMID- 11500444 TI - Enteropathogenic Escherichia coli activates ezrin, which participates in disruption of tight junction barrier function. AB - Enteropathogenic Escherichia coli (EPEC) is an important human intestinal pathogen, especially in infants. EPEC adherence to intestinal epithelial cells induces the accumulation of a number of cytoskeletal proteins beneath the bacteria, including the membrane-cytoskeleton linker ezrin. Evidence suggests that ezrin can participate in signal transduction. The aim of this study was to determine whether ezrin is activated following EPEC infection and if it is involved in the cross talk with host intestinal epithelial cells. We show here that following EPEC attachment to intestinal epithelial cells there was significant phosphorylation of ezrin, first on threonine and later on tyrosine residues. A significant increase in cytoskeleton-associated ezrin occurred following phosphorylation, suggesting activation of this molecule. Nonpathogenic E. coli and EPEC strains harboring mutations in type III secretion failed to elicit this response. Expression of dominant-negative ezrin significantly decreased the EPEC-elicited association of ezrin with the cytoskeleton and attenuated the disruption of intestinal epithelial tight junctions. These results suggest that ezrin is involved in transducing EPEC-initiated signals that ultimately affect host physiological functions. PMID- 11500445 TI - Opsonized virulent Edwardsiella tarda strains are able to adhere to and survive and replicate within fish phagocytes but fail to stimulate reactive oxygen intermediates. AB - Edwardsiella tarda is responsible for hemorrhagic septicemia (edwardsiellosis) in fish and also causes diseases in higher vertebrates such as birds, reptiles, and mammals, including humans. Interactions of E. tarda with blue gourami phagocytes were studied by light microscopy as well as by adherence, intracellular replication, and superoxide anion assays. Both nonopsonized virulent (PPD130/91 and AL9379) and avirulent (PPD125/87 and PPD76/87) bacteria could adhere to and survive and replicate within phagocytes, while only opsonized virulent strains replicated within the phagocytes. Furthermore, only avirulent E. tarda elicited a higher rate of production of reactive oxygen intermediates (ROIs) by phagocytes, indicating that they were unable to avoid and/or resist reactive oxygen radical based killing by the fish phagocytes. TnphoA transposon mutagenesis was used to construct a library of 200 alkaline phosphatase (PhoA+) fusion mutants from a total of 182,000 transconjugants derived from E. tarda PPD130/91. Five of these mutants induced more ROI production in phagocytes than the wild-type strain. Two mutants had lower replication ability inside phagocytes and moderately higher 50% lethal dose values than the wild-type strain. Sequence analysis revealed that three of these mutants had insertions at sequences having homology to PhoS, dipeptidase, and a surface polymer ligase of lipid A core proteins of other pathogens. These three independent mutations might have changed the cell surface characteristics of the bacteria, which in turn induced phagocytes to produce increased ROIs. Sequences from two other mutants had no homology to known genes, indicating that they may be novel genes for antiphagocytic killing. The present study showed that there are differences in the interactions of virulent and avirulent E. tarda organisms with fish phagocytes and PhoA+ fusion mutants that could be used successfully to identify virulence genes. The information elucidated here would help in the development of suitable strategies to combat the disease caused by E. tarda. PMID- 11500446 TI - Porphyromonas gingivalis traffics to autophagosomes in human coronary artery endothelial cells. AB - Porphyromonas gingivalis is a periodontal pathogen that also localizes to atherosclerotic plaques. Our previous studies demonstrated that P. gingivalis is capable of invading endothelial cells and that intracellular bacteria are contained in vacuoles that resemble autophagosomes. In this study, we have examined the trafficking of P. gingivalis 381 to the autophagic pathway. P. gingivalis 381 internalized by human coronary artery endothelial (HCAE) cells is located within vacuoles morphologically identical to autophagosomes. The progression of P. gingivalis 381 through intracellular vacuoles was analyzed by immunofluorescence microscopy. Vacuoles containing P. gingivalis colocalize with Rab5 and HsGsa7p early after internalization. At later times, P. gingivalis colocalizes with BiP and then progresses to a vacuole that contains BiP and lysosomal glycoprotein 120. Late endosomal markers and the lysosomal cathepsin L do not colocalize with P. gingivalis 381. The intracellular survival of P. gingivalis 381 decreases over 8 h in HCAE cells pretreated with the autophagy inhibitors 3-methyladenine and wortmannin. In addition, the vacuole containing P. gingivalis 381 lacks BiP but contains cathepsin L in the presence of wortmannin. These results suggest that P. gingivalis 381 evades the endocytic pathway to lysosomes and instead traffics to the autophagosome. PMID- 11500447 TI - Candidate vaccine against botulinum neurotoxin serotype A derived from a Venezuelan equine encephalitis virus vector system. AB - A candidate vaccine against botulinum neurotoxin serotype A (BoNT/A) was developed by using a Venezuelan equine encephalitis (VEE) virus replicon vector. This vaccine vector is composed of a self-replicating RNA containing all of the VEE nonstructural genes and cis-acting elements and also a heterologous immunogen gene placed downstream of the subgenomic 26S promoter in place of the viral structural genes. In this study, the nontoxic 50-kDa carboxy-terminal fragment (H(C)) of the BoNT/A heavy chain was cloned into the replicon vector (H(C) replicon). Cotransfection of BHK cells in vitro with the H(C)-replicon and two helper RNA molecules, the latter encoding all of the VEE structural proteins, resulted in the assembly and release of propagation-deficient, H(C) VEE replicon particles (H(C)-VRP). Cells infected with H(C)-VRP efficiently expressed this protein when analyzed by either immunofluorescence or by Western blot. To evaluate the immunogenicity of H(C)-VRP, mice were vaccinated with various doses of H(C)-VRP at different intervals. Mice inoculated subcutaneously with H(C)-VRP were protected from an intraperitoneal challenge of up to 100,000 50% lethal dose units of BoNT/A. Protection correlated directly with serum enzyme-linked immunosorbent assay titers to BoNT/A. The duration of the immunity achieved was tested at 6 months and at 1 year postvaccination, and mice challenged at these times remained refractory to challenge with BoNT/A. PMID- 11500449 TI - In vivo activation of dendritic cells and T cells during Salmonella enterica serovar Typhimurium infection. AB - The present study was initiated to gain insight into the interaction between splenic dendritic cells (DC) and Salmonella enterica serovar Typhimurium in vivo. Splenic phagocytic cell populations associated with green fluorescent protein (GFP)-expressing bacteria and the bacterium-specific T-cell response were evaluated in mice given S. enterica serovar Typhimurium expressing GFP and ovalbumin. Flow cytometry analysis revealed that GFP-positive splenic DC (CD11c+ major histocompatibility complex class II-positive [MHC-II+] cells) were present following bacterial administration, and confocal microscopy showed that GFP expressing bacteria were contained within CD11c+ MHC-II+ splenocytes. Furthermore, splenic DC and T cells were activated following Salmonella infection. This was shown by increased surface expression of CD86 and CD40 on CD11c+ MHC-II+ cells and increased CD44 and CD69 expression on CD4+ and CD8+ T cells. Salmonella-specific gamma interferon (IFN-gamma)-producing cells in both of these T-cell subsets, as well as cytolytic effector cells, were also generated in mice given live bacteria. The frequency of Salmonella-specific CD4+ T cells producing IFN-gamma was greater than that of specific CD8+ T cells producing IFN gamma in the same infected animal. This supports the argument that the predominant source of IFN-gamma production by cells of the specific immune response is CD4+ T cells. Finally, DC that phagocytosed live or heat-killed Salmonella in vitro primed bacterium-specific IFN-gamma-producing CD4+ and CD8+ T cells as well as cytolytic effector cells following administration into naive mice. Together these data suggest that DC are involved in priming naive T cells to Salmonella in vivo. PMID- 11500448 TI - Cholera toxin B subunit as a carrier molecule promotes antigen presentation and increases CD40 and CD86 expression on antigen-presenting cells. AB - Cholera toxin B subunit (CTB) is an efficient mucosal carrier molecule for the generation of mucosal antibody responses and/or induction of systemic T-cell tolerance to linked antigens. CTB binds with high affinity to GM1 ganglioside cell surface receptors. In this study, we evaluated how conjugation of a peptide or protein antigen to CTB by chemical coupling or genetic fusion influences the T cell-activating capacity of different antigen-presenting cell (APC) subsets. Using an in vitro system in which antigen-pulsed APCs were incubated with antigen specific, T-cell receptor-transgenic T cells, we found that the dose of antigen required for T-cell activation could be decreased >10,000-fold using CTB conjugated compared to free antigen. In contrast, no beneficial effects were observed when CTB was simply admixed with antigen. CTB conjugation enhanced the antigen-presenting capacity not only of dendritic cells and B cells but also of macrophages, which expressed low levels of cell surface major histocompatibility complex (MHC) class II and were normally poor activators of naive T cells. Enhanced antigen-presenting activity by CTB-linked antigen resulted in both increased T-cell proliferation and increased interleukin-12 and gamma interferon secretion and was associated with up-regulation of CD40 and CD86 on the APC surface. These results imply that conjugation to CTB dramatically lowers the threshold concentration of antigen required for immune cell activation and also permits low-MHC II-expressing APCs to prime for a specific immune response. PMID- 11500450 TI - Discrete protein determinant directs the species-specific adherence of Porphyromonas gingivalis to oral streptococci. AB - For pathogens to survive in the human oral cavity, they must identify a suitable niche in the complex multispecies biofilm that exists on oral tissues. The periodontal pathogen Porphyromonas gingivalis adheres to Streptococcus gordonii by interacting with a specific region of the streptococcal SspB polypeptide, designated BAR. However, it does not adhere to Streptococcus mutans, which expresses SpaP, a highly conserved homolog of SspB. Comparison of the predicted secondary structure of BAR with the corresponding region of SpaP suggested that the substitution of Asn for Gly1182 and Val for Pro1185 in SspB may confer a unique local structure that is not conserved in SpaP. A synthetic peptide of 26 amino acids that encompassed residues 1167 to 1193 of SspB promoted avid adherence of P. gingivalis, whereas a peptide derived from the region corresponding to BAR in SpaP was inactive. Substitution of Gly1182 and Pro1185 for Asn1182 and Val1185 in SspB by site-specific mutation generated proteins that were predicted to assume an SpaP-like secondary structure, and the purified proteins did not promote P. gingivalis adherence. Furthermore, Enterococcus faecalis strains expressing the site-specific mutants did not support adherence of P. gingivalis cells. In contrast, P. gingivalis adhered efficiently to E. faecalis strains expressing intact SspB or SspB-SpaP chimeric proteins containing BAR. These results suggest that a region of SspB consisting of 26 amino acids is sufficient to mediate the adherence of P. gingivalis to S. gordonii and that the species specificity of adherence arises from its interaction with a discrete structural determinant of SspB that is not conserved in SpaP. PMID- 11500451 TI - dksA is required for intercellular spread of Shigella flexneri via an RpoS independent mechanism. AB - Pathogenesis of Shigella flexneri is dependent on the ability of the bacterium to invade and spread within epithelial cells. In this study, we identified dksA as a gene necessary for intercellular spread in, but not invasion of, cultured cells. The S. flexneri dksA mutant exhibited sensitivity to acid and oxidative stress, in part due to an effect of DksA on production of RpoS. However, an S. flexneri rpoS mutant formed plaques on tissue culture monolayers, thus excluding DksA regulation of RpoS as the mechanism responsible for the inability of the dksA mutant to spread intercellularly. Intracellular analysis of the dksA mutant indicates that it survived and divided within the Henle cell cytoplasm, but the dksA mutant cells were elongated, and some exhibited filamentation in the intracellular environment. Some of the S. flexneri dksA mutant cells showed aberrant localization of virulence protein IcsA, which may inhibit spread between epithelial cells. PMID- 11500452 TI - Cytolethal distending toxin demonstrates genotoxic activity in a yeast model. AB - Cytolethal distending toxins (CDTs) are multisubunit proteins produced by a variety of bacterial pathogens that cause enlargement, cell cycle arrest, and apoptosis in mammalian cells. While their function remains uncertain, recent studies suggest that they can act as intracellular DNases in mammalian cells. Here we establish a novel yeast model for understanding CDT-associated disease. Expression of the CdtB subunit in yeast causes a G2/M arrest, as seen in mammalian cells. CdtB toxicity is not circumvented in yeast genetically altered to lack DNA damage checkpoint control or that constitutively promote cell cycle progression via mutant Cdk1, because CdtB causes a permanent type of damage that results in loss of viability. Finally, we establish that CDTs are likely to be potent genotoxins, as indicated by in vivo degradation of chromosomal DNA associated with expression of CdtB-suggesting that the varied distribution of CDT in bacteria implicates many human pathogens as possessors of genotoxic activity. PMID- 11500453 TI - Detection of melanin-like pigments in the dimorphic fungal pathogen Paracoccidioides brasiliensis in vitro and during infection. AB - Melanins are implicated in the pathogenesis of several human diseases, including some microbial infections. In this study, we analyzed whether the conidia and the yeasts of the thermally dimorphic fungal pathogen Paracoccidioides brasiliensis produce melanin or melanin-like compounds in vitro and during infection. Growth of P. brasiliensis mycelia on water agar alone produced pigmented conidia, and growth of yeasts in minimal medium with L-3,4-dihydroxyphenylalanine (L-DOPA) produced pigmented cells. Digestion of the pigmented conidia and yeasts with proteolytic enzymes, denaturant, and hot concentrated acid yielded dark particles that were the same size and shape as their propagules. Immunofluorescence analysis demonstrated reactivity of a melanin-binding monoclonal antibody (MAb) with the pigmented conidia, yeasts, and particles. Electron spin resonance spectroscopy identified the yeast-derived particles produced in vitro when P. brasiliensis was grown in L-DOPA medium as a melanin-like compound. Nonreducing polyacrylamide gel electrophoresis of cytoplasmic yeast extract revealed a protein that catalyzed melanin synthesis from L-DOPA. The melanin binding MAb reacted with yeast cells in tissue from mice infected with P. brasiliensis. Finally digestion of infected tissue liberated particles reactive to the melanin binding MAb that had the typical morphology of P. brasiliensis yeasts. These data strongly suggest that P. brasiliensis propagules, both conidia and yeast cells, can produce melanin or melanin-like compounds in vitro and in vivo. Based on what is known about the function of melanin in the virulence of other fungi, this pigment may play a role in the pathogenesis of paracoccidioidomycosis. PMID- 11500454 TI - Fas/Fas ligand system mediates epithelial injury, but not pulmonary host defenses, in response to inhaled bacteria. AB - The Fas/Fas ligand (FasL) system has been implicated in alveolar epithelial cell apoptosis during pulmonary fibrosis and acute respiratory distress syndrome. However, Fas ligation can also lead to cell activation and cytokine production. The goal of this study was to determine the role of the Fas/FasL system in host defenses against Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae. We administered bacteria by aerosolization into the lungs of Fas deficient (lpr) mice and wild-type (C57BL/6) mice and measured bacterial clearance at 6 and 12 h. One hour prior to euthanasia, the mice received an intraperitoneal injection of human serum albumin (HSA) for alveolar permeability determinations. At all times after bacterial challenges, the lungs of the lpr mice contained similar or lower numbers of bacteria than those of the C57BL/6 mice. Alveolar permeability changes, as determined by bronchoalveolar lavage fluid HSA concentrations, were less severe in the lpr mice 6 h after the challenges. In response to E. coli, the lpr mice had significantly more polymorphonuclear leukocytes (PMN) and macrophage inflammatory protein 2 in the lungs, whereas histopathologic changes were less severe. In contrast, in response to the gram-positive cocci, the lpr animals had similar or lower numbers of PMN. We conclude that the Fas/FasL system contributes to the development of permeability changes and tissue injury during-gram negative bacterial pneumonia. The Fas/FasL system did not have a major role in the clearance of aerosolized bacteria from the lungs at the bacterial doses tested. PMID- 11500455 TI - Identification and characterization of mycobacterial proteins differentially expressed under standing and shaking culture conditions, including Rv2623 from a novel class of putative ATP-binding proteins. AB - The environmental signals that affect gene regulation in Mycobacterium tuberculosis remain largely unknown despite their importance to tuberculosis pathogenesis. Other work has shown that several promoters, including acr (also known as hspX) (alpha-crystallin homolog), are upregulated in shallow standing cultures compared with constantly shaking cultures. Each of these promoters is also induced to a similar extent within macrophages. The present study used two dimensional gel electrophoresis and mass spectrometry to further characterize differences in mycobacterial protein expression during growth under standing and shaking culture conditions. Metabolic labeling of M. bovis BCG showed that at least 45 proteins were differentially expressed under standing and shaking culture conditions. Rv2623, CysA2-CysA3, Gap, and Acr were identified from each of four spots or gel bands that were specifically increased in bacteria from standing cultures. An additional standing-induced spot contained two comigrating proteins, GlcB and KatG. The greatest induction was observed with Rv2623, a 32 kDa protein of unknown function that was strongly expressed under standing conditions and absent in shaking cultures. Analysis using PROBE, a multiple sequence alignment and database mining tool, classified M. tuberculosis Rv2623 as a member of a novel class of ATP-binding proteins that may be involved in M. tuberculosis's response to environmental signals. These studies demonstrate the power of combined proteomic and computational approaches and demonstrate that subtle differences in bacterial culture conditions may have important implications for the study of gene expression in mycobacteria. PMID- 11500456 TI - Towards development of an edible vaccine against bovine pneumonic pasteurellosis using transgenic white clover expressing a Mannheimia haemolytica A1 leukotoxin 50 fusion protein. AB - Development of vaccines against bovine pneumonia pasteurellosis, or shipping fever, has focused mainly on Mannheimia haemolytica A1 leukotoxin (Lkt). In this study, the feasibility of expressing Lkt in a forage plant for use as an edible vaccine was investigated. Derivatives of the M. haemolytica Lkt in which the hydrophobic transmembrane domains were removed were made. Lkt66 retained its immunogenicity and was capable of eliciting an antibody response in rabbits that recognized and neutralized authentic Lkt. Genes encoding a shorter Lkt derivative, Lkt50, fused to a modified green fluorescent protein (mGFP5), were constructed for plant transformation. Constructs were screened by Western immunoblot analysis for their ability to express the fusion protein after agroinfiltration in tobacco. The fusion construct pBlkt50-mgfp5, which employs the cauliflower mosaic virus 35S promoter for transcription, was selected and introduced into white clover by Agrobacterium tumefaciens-mediated transformation. Transgenic lines of white clover were recovered, and expression of Lkt50-GFP was monitored and confirmed by laser confocal microscopy and Western immunoblot analysis. Lkt50-GFP was found to be stable in clover tissue after drying of the plant material at room temperature for 4 days. An extract containing Lkt50-GFP from white clover was able to induce an immune response in rabbits (via injection), and rabbit antisera recognized and neutralized authentic Lkt. This is the first demonstration of the expression of an M. haemolytica antigen in plants and paves the way for the development of transgenic plants expressing M. haemolytica antigens as an edible vaccine against bovine pneumonic pasteurellosis. PMID- 11500457 TI - Mutualism versus independence: strategies of mixed-species oral biofilms in vitro using saliva as the sole nutrient source. AB - During initial dental plaque formation, the ability of a species to grow when others cannot would be advantageous, and enhanced growth through interspecies and intergeneric cooperation could be critical. These characteristics were investigated in three coaggregating early colonizers of the tooth surface (Streptococcus gordonii DL1, Streptococcus oralis 34, and Actinomyces naeslundii T14V). Area coverage and cell cluster size measurements showed that attachment of A. naeslundii and of S. gordonii to glass flowcells was enhanced by a salivary conditioning film, whereas attachment of S. oralis was hindered. Growth experiments using saliva as the sole carbon and nitrogen source showed that A. naeslundii was unable to grow either in planktonic culture or as a biofilm, whereas S. gordonii grew under both conditions. S. oralis grew planktonically, but to a much lower maximum cell density than did S. gordonii; S. oralis did not grow reproducibly as a biofilm. Thus, only S. gordonii possessed all traits advantageous for growth as a solitary and independent resident of the tooth. Two species biofilm experiments analyzed by laser confocal microscopy showed that neither S. oralis nor A. naeslundii grew when coaggregated pairwise with S. gordonii. However, both S. oralis and A. naeslundii showed luxuriant, interdigitated growth when paired together in coaggregated microcolonies. Thus, the S. oralis-A. naeslundii pair formed a mutualistic relationship, potentially contact dependent, that allows each to grow where neither could survive alone. S. gordonii, in contrast, neither was hindered by nor benefited from the presence of either of the other strains. The formation of mutually beneficial interactions within the developing biofilm may be essential for certain initial colonizers to be retained during early plaque development, whereas other initial colonizers may be unaffected by neighboring cells on the substratum. PMID- 11500458 TI - Type 3 fimbrial shaft (MrkA) of Klebsiella pneumoniae, but not the fimbrial adhesin (MrkD), facilitates biofilm formation. AB - Isolates of Klebsiella pneumoniae are responsible for opportunistic infections, particularly of the urinary tract and respiratory tract, in humans. These bacteria express type 3 fimbriae that have been implicated in binding to eucaryotic cells and matrix proteins. The type 3 fimbriae mediate binding to target tissue using the MrkD adhesin that is associated with the fimbrial shaft comprised of the MrkA protein. The formation of biofilms in vitro by strains of K. pneumoniae was shown to be affected by the production of fimbriae on the bacterial surface. However, a functional MrkD adhesin was not necessary for efficient biofilm formation. Nonfimbriate strains were impaired in their ability to form biofilms. Using isogenic fimbriate and nonfimbriate strains of K. pneumoniae expressing green fluorescent protein it was possible to demonstrate that the presence of type 3 fimbriae facilitated the formation of dense biofilms in a continuous-flowthrough chamber. Transformation of nonfimbriate mutants with a plasmid possessing an intact mrk gene cluster restored the fimbrial phenotype and the rapid ability to form biofilms. PMID- 11500459 TI - Filarial antigens impair the function of human dendritic cells during differentiation. AB - The antigen-specific T-cell unresponsiveness seen in lymphatic filariasis is mediated, in part, by diminished antigen-presenting cell function and is most specific for microfilariae (MF), the parasite stage found in large numbers in the peripheral circulation. We investigated the effect of MF antigen (MFAg) on dendritic cells (DC) in both their differentiation process from monocyte precursors and also after they have developed into DC. When MFAg was added to cultures of monocytes during their differentiation process to immature DC, the production of interleukin 12 (IL-12) p40, p70 protein, and IL-10 was significantly (P < 0.03) inhibited in response to Staphylococcus aureus Cowan (SAC) and SAC-gamma interferon (IFN-gamma) (60% to 80% inhibition). IL-10 was also inhibited (P = 0.04) in response to CD40 ligand-IFN-gamma. Moreover, MFAg inhibited the mRNA expression of IL-12 p40 and IL-10 as assessed by RNA protection assays. This effect was antigen specific, as another parasite antigen (soluble Toxoplasma gondii antigen) did not inhibit the production of these cytokines. This effect was also not a result of diminished cell viability nor of an alteration in surface expression of most costimulatory surface molecules, including major histocompatibility complex class I and class II. In contrast to exposure throughout the differentiation process, MFAg added to immature DC had no effect on DC cytokine expression. Although MF-differentiated DC were capable of inducing an allogeneic mixed lymphocyte reaction, they did so to a significantly lesser degree than DC without antigen exposure. These data collectively suggest that once DC are differentiated from their precursor cells, they become resistant to changes by MFAg. PMID- 11500460 TI - Effects of Mycobacterium bovis BCG infection on regulation of L-arginine uptake and synthesis of reactive nitrogen intermediates in J774.1 murine macrophages. AB - The generation of nitric oxide (NO) by activated macrophages is believed to control mycobacterial infection in the murine system. In this study we examined the effect of Mycobacterium bovis BCG infection on the L-arginine-dependent NO pathway in J774.1 murine macrophages. We have confirmed previous results by demonstrating that stimulation of J774.1 with lipopolysaccharide (LPS) and gamma interferon (IFN-gamma) results in an increase in the uptake of 3H-labeled L arginine and a concomitant increase in the production of NO. We have also shown that BCG can mimic LPS treatment, leading to enhanced L-[3H]arginine uptake by IFN-gamma-stimulated macrophages. Lipoarabinomannan, a component of the BCG cell wall that is structurally similar to LPS, is not responsible for the uptake stimulation in IFN-gamma stimulated macrophages. Although we demonstrated that there was a 2.5-fold increase in NO production by macrophages 4 h after LPS-IFN gamma stimulation, BCG infection (with or without IFN-gamma stimulation) did not lead to the production of NO by the macrophages by 4 h postinfection. At 24 h postinfection, the infected macrophages that were stimulated with IFN-gamma produced amounts of NO similar to those of macrophages stimulated with LPS-IFN gamma. This suggests that there are multiple regulatory pathways involved in the production of NO. Finally, our data suggest that increased expression of the arginine permease, MCAT2B, after 4 h of LPS-IFN-gamma treatment or BCG infection IFN-gamma treatment is not sufficient to account for the increases in L [3H]arginine uptake detected. This suggests that the activity of the L-arginine transporter(s) is also altered in response to macrophage activation. PMID- 11500461 TI - Delayed invasion of the kidney and brain by Borrelia crocidurae in plasminogen deficient mice. AB - Borrelia crocidurae is an etiologic agent of relapsing fever in Africa and is transmitted to humans by the bite of soft ticks of the genus Ornithodoros. The role of the plasminogen (Plg) activation system for the pathogenicity of B. crocidurae was investigated by infection of Plg-deficient (plg(-/-)) and Plg wild type (plg(+/+)) mice. No differences in spirochetemia were observed between the plg(-/-) and plg(+/+) mice. However, signs indicative of brain invasion, such as neurological symptoms and/or histopathological changes, were more common in plg(+/+) mice. Quantitative immunohistochemical analysis demonstrated infection of spirochetes in kidney interstitium and brain as soon as 2 days postinoculation. Lower numbers of extravascular spirochetes in plg(-/-) mice during the first days of infection suggested a less efficient invasion mechanism in these mice than in the plg(+/+) mice. The invasion of the kidneys in plg(-/-) mice produced no significant inflammation, as seen by quantitative immunohistochemistry of the CD45 common leukocyte marker. However, significant kidney inflammation was observed with infection in the plg(+/+) mice. In brain, inflammation was more severe in plg(+/+) mice than in plg(-/-) mice, and the numbers of CD45(+) cells increased significantly with duration of infection in the plg(+/+) mice. The results show that invasion of brain and kidney occurs as early as 2 days after inoculation. Also, Plg is not required for establishment of spirochetemia by the organism, whereas it is involved in the invasion of organs. PMID- 11500462 TI - Distinct proinflammatory host responses to Neisseria gonorrhoeae infection in immortalized human cervical and vaginal epithelial cells. AB - In this study we utilized immortalized morphologically and functionally distinct epithelial cell lines from normal human endocervix, ectocervix, and vagina to characterize gonococcal epithelial interactions pertinent to the lower female genital tract. Piliated, but not nonpiliated, N. gonorrhoeae strain F62 variants actively invaded these epithelial cell lines, as demonstrated by an antibiotic protection assay and confocal microscopy. Invasion of these cells by green fluorescent protein-expressing gonococci was characterized by colocalization of gonococci with F actin, which were initially detected 30 min postinfection. In all three cell lines, upregulation of interleukin 8 (IL-8) and IL-6, intercellular adhesion molecule 1 (CD54), and the nonspecific cross-reacting antigen (CD66c) were detected 4 h after infection with piliated and nonpiliated gonococci. Furthermore, stimulation of all three cell lines with gonococcal whole cell lysates resulted in a similar upregulation of IL-6 and IL-8, confirming that bacterial uptake is not essential for this response. Increased levels of IL-1 were first detected 8 h after infection with gonococci, suggesting that the earlier IL-8 and IL-6 responses were not mediated through the IL-1 signaling pathway. The IL-1 response was limited to cultures infected with piliated gonococci and was more vigorous in the endocervical epithelial cells. The ability of gonococci to stimulate distinct proinflammatory host responses in these morphologically and functionally different compartments of the lower female genital tract may contribute directly to the inflammatory signs and symptoms characteristic of disease caused by N. gonorrhoeae. PMID- 11500463 TI - Fresh isolates from children with severe Plasmodium falciparum malaria bind to multiple receptors. AB - The sequestration of Plasmodium falciparum-infected erythrocytes (pRBC) away from the peripheral circulation is a property of all field isolates. Here we have examined the pRBC of 111 fresh clinical isolates from children with malaria for a number of adhesive features in order to study their possible coexpression and association with severity of disease. A large number of adhesion assays were performed studying rosetting, giant rosetting, and binding to CD36, intercellular adhesion molecule 1, platelet endothelial cell adhesion molecule 1, thrombospondin, heparin, blood group A, and immunoglobulins. Suspension assays were performed at the actual parasitemia of the isolate, while all the static adhesion assays were carried out at an equal adjusted parasitemia. The ability to bind to multiple receptors, as well as the ability to form rosettes and giant rosettes, was found to be more frequent among isolates from children with severe versus mild malaria (P = 0.0015). Rosettes and giant rosettes were more frequent for children with severe malaria, and the cell aggregates were larger and tighter, than for those with mild disease (P = 0.0023). Binding of immunoglobulins (97% of isolates) and of heparin (81% of isolates) to infected erythrocytes was common, and binding to heparin and blood group A was associated with severity of disease (P = 0.011 and P = 0.031, respectively). These results support the idea that isolates that bind to multiple receptors are involved in the causation of severe malaria and that several receptor-ligand interactions work synergistically in bringing about severe disease. PMID- 11500464 TI - In vivo and in vitro studies of cytosolic phospholipase A2 expression in Helicobacter pylori infection. AB - Modifications of mucosal phospholipids have been detected in samples from patients with Helicobacter pylori-positive gastritis. These alterations appear secondary to increased phospholipase A2 activity (PLA2). The cytosolic form of this enzyme (cPLA2), normally involved in cellular signaling and growth, has been implicated in cancer pathogenesis. The aim of this study was to investigate cPLA2 expression and PLA2 activity in the gastric mucosae of patients with and without H. pylori infection. In gastric biopsies from 10 H. pylori-positive patients, cPLA2 levels, levels of mRNA as determined by reverse transcriptase PCR, levels of protein as determined by immunohistochemistry, and total PLA2 activity were higher than in 10 H. pylori-negative gastritis patients. To clarify whether H. pylori had a direct effect on the cellular expression of cPLA2, we studied cPLA2 expression in vitro with different human epithelial cell lines, one from a patient with larynx carcinoma (i.e., HEp-2 cells) and two from patients with gastric adenocarcinoma (i.e., AGS and MKN 28 cells), incubated with different H. pylori strains. The levels of cPLA2, mRNA, and protein expression were unchanged in Hep-2 cells independently of cellular adhesion or invasion of the bacteria. Moreover, no change in cPLA2 protein expression was observed in AGS or MKN 28 cells treated with wild-type H. pylori. In conclusion, our study shows increased cPLA2 expression and PLA2 activity in the gastric mucosae of patients with H. pylori infection and no change in epithelial cell lines exposed to H. pylori. PMID- 11500465 TI - Gene cluster for assembly of pilus colonization factor antigen III of enterotoxigenic Escherichia coli. AB - The assembly of pilus colonization factor antigen III (CFA/III) of enterotoxigenic Escherichia coli (ETEC) requires the processing of CFA/III major pilin (CofA) by a prepilin peptidase (CofP), similar to other type IV pilus formation systems. CofA is produced initially as a 26.5-kDa preform pilin (prepilin) and then processed to a 20.5-kDa mature pilin by CofP which is predicted to be localized in the inner membrane. In the present experiment, we determined the nucleotide sequence of the whole region for CFA/III formation and identified a cluster of 14 genes, including cofA and cofP. Several proteins encoded by cof genes were similar to previously described proteins, such as the toxin-coregulated pili of Vibrio cholerae and the bundle-forming pili of enteropathogenic E. coli. The G+C content of the cof gene cluster was 37%, which was significantly lower than the average for the E. coli genome (50%). The introduction of a recombinant plasmid containing the cof gene cluster into the E. coli K-12 strain conferred CFA/III biogenesis and the ability of adhesion to the human colon carcinoma cell line Caco-2. This is the first report of a complete nucleotide sequence of the type IV pili found in human ETEC, and our results provide a useful model for studying the molecular mechanism of CFA/III biogenesis and the role of CFA/III in ETEC infection. PMID- 11500466 TI - HLA class II polymorphism influences onset and severity of pathology in Schistosoma mansoni-infected transgenic mice. AB - Genetic factors that might influence susceptibility or resistance in naive individuals and early-stage pathology in schistosomiasis are difficult to study in clinical trials, since in areas where the disease is endemic the first contact with the parasite occurs most often at very early ages. Therefore, four strains (DR1.Abeta degrees, DR2.Abeta degrees, DQ8.Abeta degrees, and DQ6.Abeta degrees ) of major histocompatibility complex class II-deficient mice (Abeta degrees ), transgenic for different HLA alleles, have been used to evaluate the potential role of HLA class II polymorphism in the onset of the infection by Schistosoma mansoni. The survival rates and parasitological and immunological parameters after infection were evaluated and compared against the control values obtained with Abeta degrees mice. All four mouse strains used in this study were able to generate a specific immune response against S. mansoni antigens (cytokine production and antibody production). However, only mice expressing DR alleles survived until the chronic stage of the infection and were able to mount protective granulomatous response avoiding hepatic damage, presenting predominant gamma interferon production. In contrast, strains expressing DQ alleles revealed an impairment in generating effective granulomas, resulting in earlier death, which was associated with an impaired hepatic granulomatous response and liquefactic necrosis, reflecting the influence of HLA polymorphism in the establishment of protective response in the early stage of infection. PMID- 11500467 TI - Dietary glycine prevents peptidoglycan polysaccharide-induced reactive arthritis in the rat: role for glycine-gated chloride channel. AB - Peptidoglycan polysaccharide (PG-PS) is a primary structural component of bacterial cell walls and causes rheumatoid-like arthritis in rats. Recently, glycine has been shown to be a potential immunomodulator; therefore, the purpose of this study was to determine if glycine would be protective in a PG-PS model of arthritis in vivo. In rats injected with PG-PS intra-articularly, ankle swelling increased 21% in 24 to 48 h and recovered in about 2 weeks. Three days prior to reactivation with PG-PS given intravenously (i.v.), rats were divided into two groups and fed a glycine-containing or nitrogen-balanced control diet. After i.v. PG-PS treatment joint swelling increased 2.1 +/- 0.3 mm in controls but only 1.0 +/- 0.2 mm in rats fed glycine. Infiltration of inflammatory cells, edema, and synovial hyperplasia in the joint were significantly attenuated by dietary glycine. Tumor necrosis factor alpha (TNF-alpha) mRNA was detected in ankle homogenates from rats fed the control diet but not in ankles from rats fed glycine. Moreover, intracellular calcium was increased significantly in splenic macrophages treated with PG-PS; however, glycine blunted the increase about 50%. The inhibitory effect of glycine was reversed by low concentrations of strychnine or chloride-free buffer, and it increased radiolabeled chloride influx nearly fourfold, an effect also inhibited by strychnine. In isolated splenic macrophages, glycine blunted translocation of the p65 subunit of NF-kappaB into the nucleus, superoxide generation, and TNF-alpha production caused by PG-PS. Further, mRNA for the beta subunit of the glycine receptor was detected in splenic macrophages. This work supports the hypothesis that glycine prevents reactive arthritis by blunting cytokine release from macrophages by increasing chloride influx via a glycine-gated chloride channel. PMID- 11500468 TI - Intermediate subunit of the Gal/GalNAc lectin of Entamoeba histolytica is a member of a gene family containing multiple CXXC sequence motifs. AB - Killing by Entamoeba histolytica requires parasite adherence to host galactose- and N-acetyl-D-galactosamine (Gal/GalNAc)-containing cell surface receptors. A 260-kDa heterodimeric E. histolytica Gal/GalNAc lectin composed of heavy (Hgl) and light (Lgl) subunits has been previously described. Here we present the cloning and characterization of Igl, a 150-kDa intermediate subunit of the Gal/GalNAc lectin. Igl, Hgl, and Lgl colocalized on the surface membrane of trophozoites. Two unlinked copies of genes encoding Igl shared 81% amino acid sequence identity (GenBank accession no. AF337950 and AF337951). They encoded cysteine-rich proteins with amino- and carboxy-terminal hydrophobic signal sequences characteristic of glycosylphosphatidylinositol (GPI)-anchored membrane proteins. The igl genes lacked carbohydrate recognition domains but were members of a large family of amebic genes containing CXXC and CXC motifs. These data indicate that Igl is part of the parasite's multimolecular Gal/GalNAc adhesin required for host interaction. PMID- 11500469 TI - Isolation and characterization of a mutant Chinese hamster ovary cell line that is resistant to Chlamydia trachomatis infection at a novel step in the attachment process. AB - Host factors involved in Chlamydia trachomatis pathogenesis were investigated by random chemical mutagenesis of Chinese hamster ovary (CHO-K1) cells followed by selection for clones resistant to chlamydial infection. A clonal mutant cell line, D4.1-3, refractory to infection by the C. trachomatis L2 serovar was isolated. The D4.1-3 cell line appears to be lacking in a previously undescribed temperature-dependent and heparin-resistant binding step that occurs subsequent to engagement of cell surface heparan sulfate by L2 elementary bodies. This novel binding step differentiates the lymphogranuloma venereum (LGV) serovar from other serovars and may contribute the different pathologies associated with LGV and non LGV strains. PMID- 11500470 TI - Proteomics reveals open reading frames in Mycobacterium tuberculosis H37Rv not predicted by genomics. AB - Genomics revealed the sequence of 3924 genes of the H37Rv strain of Mycobacterium tuberculosis. Proteomics complements genomics in showing which genes are really expressed, and here we show the expression of six genes not predicted by genomics, as proved by two-dimensional electrophoresis and matrix-assisted laser desorption ionization and nano-electrospray mass spectrometry. PMID- 11500471 TI - PcrV immunization enhances survival of burned Pseudomonas aeruginosa-infected mice. AB - Burned Pseudomonas aeruginosa-infected mice immunized against PcrV, a type III virulence system translocating protein, showed significantly enhanced survival compared to controls. Survival was non-O serotype specific and correlated with a reduced systemic microbial load. Infection with a high-level toxin A-producing strain required supplemental antitoxin treatment to enhance survival. PMID- 11500472 TI - Brucella abortus HtrA functions as an authentic stress response protease but is not required for wild-type virulence in BALB/c mice. AB - A second mutation has recently been identified in the previously described Brucella abortus htrA mutant PHE1. As a result of this finding, a new B. abortus htrA mutant, designated RWP11, was constructed to evaluate the biological function of the Brucella HtrA protease. RWP11 is more sensitive to oxidative killing in vitro and less resistant to killing by cultured murine neutrophils and macrophages than the virulent parental strain 2308 but is not attenuated in BALB/c mice through 4 weeks postinfection. The in vitro phenotype of B. abortus RWP11 is consistent with the proposed function of bacterial HtrA proteases as components of a secondary line of defense against oxidative damage. The in vivo phenotype of this mutant, however, indicates that, unlike the corresponding Salmonella and Yersinia proteins, Brucella HtrA does not play a critical role in virulence in the mouse model. PMID- 11500473 TI - Cloning, expression, and catalytic activity of Helicobacter hepaticus urease. AB - Helicobacter hepaticus causes disease in the liver and lower intestinal tract of mice. It is strongly urease positive, although it does not live in an acidic environment. The H. hepaticus urease gene cluster was expressed in Escherichia coli with and without coexpression of the Helicobacter pylori nickel transporter NixA. As for H. pylori, it was difficult to obtain enzymatic activity from recombinant H. hepaticus urease; special conditions including NiCl2 supplementation were required. The H. hepaticus urease cluster contains a homolog of each gene in the H. pylori urease cluster, including the urea transporter gene ureI. Downstream genes were homologs of the nik nickel transport operon of E. coli. Nongastric H. hepaticus produces urease similar to that of H. pylori. PMID- 11500474 TI - Demonstration of polysaccharide capsule in Campylobacter jejuni using electron microscopy. AB - Recently, we reported that Campylobacter jejuni, an important gastrointestinal pathogen, has the genetic determinants to produce a capsular polysaccharide (Karlyshev et al., Mol. Microbiol. 35:529-541, 2000). Despite these data, the presence of a capsule in these bacteria has remained controversial. In this study we stain C. jejuni cells with the cationic dye Alcian blue and demonstrate for the first time by electron microscopy that C. jejuni cells produce a polysaccharide capsule that is retained in the coccoid form but is absent in a kpsM mutant. PMID- 11500475 TI - Recombinant Actinobacillus actinomycetemcomitans cytolethal distending toxin proteins are required to interact to inhibit human cell cycle progression and to stimulate human leukocyte cytokine synthesis. AB - It has recently been discovered that Actinobacillus actinomycetemcomitans, an oral bacterium causing periodontitis, produces cytolethal distending toxin (CDT), a cell cycle-modulating toxin that has three protein subunits: CdtA, CdtB, and CdtC. In this study, we have cloned and expressed each toxin gene from A. actinomycetemcomitans in Escherichia coli and purified the recombinant Cdt proteins to homogeneity. Individual Cdt proteins failed to induce cell cycle arrest of the human epithelial cell line HEp-2. The only combinations of toxin proteins causing cell cycle arrest were the presence of all three Cdt proteins and the combination of CdtB and CdtC. A similar experimental protocol was used to determine if recombinant Cdt proteins were able to induce human peripheral blood mononuclear cells (PBMCs) to produce cytokines. The individual Cdt proteins were able to induce the synthesis by PBMCs of interleukin-1beta (IL-1beta), IL-6, and IL-8 but not of tumor necrosis factor alpha, IL-12, or granulocyte-macrophage colony-stimulating factor, with CdtC being the most potent and CdtB being the least potent cytokine inducer. There was evidence of synergism between these Cdt proteins in the stimulation of cytokine production, most markedly with gamma interferon, which required the minimum interaction of CdtB and -C to stimulate production. PMID- 11500476 TI - Escherichia coli cytotoxic necrotizing factor and Pasteurella multocida toxin induce focal adhesion kinase autophosphorylation and Src association. AB - Cytotoxic necrotizing factor 1 and Pasteurella multocida toxin induced dose- and time-dependent increases in focal adhesion kinase (FAK) Tyr397 phosphorylation in Swiss 3T3 cells. FAK autophosphorylation was sensitive to inhibitors of p160/ROCK and coincided with the formation of stable complexes between FAK and Src family members. PMID- 11500477 TI - Characterization of the secreted MPT53 antigen of Mycobacterium tuberculosis. AB - MPT53 is a secreted protein of Mycobacterium tuberculosis. Southern transfer and hybridization showed mpt53 to be conserved in the M. tuberculosis complex and to have homology with DNA from Mycobacterium avium and other nontuberculous mycobacteria. However, anti-MPT53 polyclonal antibodies detected no antigen in the culture filtrates of M. avium and other nontuberculous mycobacteria. MPT53 of M. tuberculosis induced strong, tuberculosis-specific antibody responses in guinea pigs but induced no delayed-type hypersensitivity. Involvement in immune responses during human tuberculosis was very modest. PMID- 11500478 TI - Cryptosporidium parvum infection requires host cell actin polymerization. AB - The intracellular protozoan parasite Cryptosporidium parvum accumulates host cell actin at the interface between the parasite and the host cell cytoplasm. Here we show that the actin polymerizing proteins Arp2/3, vasodilator-stimulated phosphoprotein (VASP), and neural Wiskott Aldrich syndrome protein (N-WASP) are present at this interface and that host cell actin polymerization is necessary for parasite infection. PMID- 11500479 TI - Isolation and characterization of a Vibrio vulnificus mutant deficient in both extracellular metalloprotease and cytolysin. AB - We isolated a Vibrio vulnificus mutant that was deficient in both metalloprotease and cytolysin by allelic exchange. The virulence of this mutant in mice and its cytotoxicity for HEp-2 cells were comparable to those of the wild-type strain, indicating that neither factor was essential for these properties. The cytolysin, but not the protease, seemed to be important for causing damage in the alimentary tract of the mice. PMID- 11500480 TI - Role of endogenous interleukin-12 in immune response to staphylococcal enterotoxin B in mice. AB - In the present study, the roles of interleukin 12 (IL-12) and IL-18 and their possible interaction during superantigen-induced responses were studied by injection of staphylococcal enterotoxin B (SEB) into mice. These data suggest that the role of IL-12 in SEB-induced responses is limited to sustaining gamma interferon release by an IL-18-independent mechanism. PMID- 11500481 TI - Development of a competitive index assay to evaluate the virulence of Listeria monocytogenes actA mutants during primary and secondary infection of mice. AB - We developed a competitive index assay for murine listeriosis that tests the virulence of Listeria monocytogenes strains in different organs and at various times postinoculation. Studies presented here demonstrate the reproducibility of this assay during primary and secondary infection of inbred and outbred mice. We verified the validity of this assay by performing competitive index analysis of a well-characterized strain of L. monocytogenes lacking the actA gene. In addition, we found that while L. monocytogenes strains unable to recruit vasodilator stimulated phosphoprotein (VASP) to their surface exhibit a 10-fold virulence attenuation in the livers of naive animals, they display a 50-fold survival defect in the liver during secondary listeriosis. PMID- 11500482 TI - Schistosome calcium channel beta subunits. Unusual modulatory effects and potential role in the action of the antischistosomal drug praziquantel. AB - Schistosomes are parasitic flatworms that cause schistosomiasis, a major tropical disease. The current drug of choice against schistosomiasis is praziquantel (PZQ), which has minimal side effects and is potent against all schistosome species. The mode of action of PZQ is unknown, though the drug clearly affects Ca(2+) homeostasis in worms, and there is indirect evidence for interaction of PZQ with schistosome voltage-gated Ca(2+) channels. We have cloned and expressed two Ca(2+) channel beta subunits, one from Schistosoma mansoni and one from Schistosoma japonicum. These two subunits (SmCa(v)beta A and SjCa(v)beta) have structural motifs that differ from those found in other known beta subunits. Surprisingly, coexpression of either SmCa(v)beta A or SjCa(v)beta with a cnidarian (CyCa(v)1) or mammalian (Ca(v)2.3) Ca(2+) channel alpha(1) subunit results in a striking reduction in current amplitude. In the case of Ca(v)2.3, this current reduction can be partially reversed by addition of 100 nm PZQ, which results in a significant increase in current amplitude. Thus, these unusual schistosome beta subunits can confer PZQ sensitivity to an otherwise PZQ insensitive mammalian Ca(2+) channel, indicating that a possible target for PZQ action is the interaction between beta subunits and pore-forming alpha(1) subunits in schistosomes. PMID- 11500483 TI - The mammalian target of rapamycin regulates C2C12 myogenesis via a kinase independent mechanism. AB - Rapamycin inhibits differentiation of mouse C2C12 myoblasts, a tissue culture model for skeletal muscle differentiation. The mechanism by which a rapamycin sensitive signaling pathway regulates myogenesis is largely unknown. The mammalian target of rapamycin (mTOR) is a central regulator of cell growth and proliferation, but its role in myogenesis has not been examined directly. Here we report the investigation of the function of mTOR and its downstream effectors in muscle differentiation. Rapamycin exerts an inhibitory effect on C2C12 myogenesis at different stages, implying that a rapamycin-sensitive pathway may be required for multiple processes during muscle differentiation. The mTOR protein level increases 10-fold during differentiation, via a post-transcriptional mechanism. As the first direct demonstration of the essential role of mTOR in muscle differentiation, we show that a rapamycin-resistant mTOR, but not S6 kinase 1, can rescue rapamycin-inhibited myogenesis. Remarkably, the myogenic function of mTOR does not require its kinase activity. Two downstream effectors of the rapamycin-sensitive pathway, S6 kinase 1 and eIF4E-binding protein 1, undergo differential regulation during myogenesis, but neither protein is the relevant effector for the myogenic signaling of mTOR. Taken together, our observations suggest a novel mTOR signaling mechanism essential for skeletal muscle differentiation. PMID- 11500484 TI - Coupling of RYR1 and L-type calcium channels via calmodulin binding domains. AB - In skeletal muscle the L-type Ca2+ channel directly controls the opening of the sarcoplasmic reticulum Ca2+ release channel (RYR1), and RYR1, in turn, prevents L type Ca2+ channel inactivation. We demonstrate that the two proteins interact using calmodulin binding regions of both proteins. A recombinant protein representing amino acids 1393-1527 (D1393-1527) of the carboxyl-terminal tail of the skeletal muscle L-type voltage-dependent calcium channel binds Ca2+, Ca2+ calmodulin, and apocalmodulin. In the absence of calmodulin, D1393-1527 binds to both RYR1 and a peptide representing the calmodulin binding site of RYR1 (amino acids 3609-3643). In addition, biotinylated R3609-3643 peptide can be used with streptavidin beads to pull down [3H]PN200-110-labeled L-type channels from detergent-solubilized transverse tubule membranes. The binding of the L-type channel carboxyl-terminal tail to the calmodulin binding site on RYR1 may stabilize the contact between the two proteins, provide a mechanism for Ca2+ and/or calmodulin regulation of their interaction, or participate directly in functional signaling between these two proteins. A unique aspect of this study is the finding that calmodulin binding sequences can serve as specific binding motifs for proteins other than calmodulin. PMID- 11500485 TI - Tyrosine phosphorylation of villin regulates the organization of the actin cytoskeleton. AB - We have previously shown that tyrosine phosphorylation of the actin-regulatory protein villin is accompanied by the redistribution of phosphorylated villin and a concomitant decrease in the F-actin content of intestinal epithelial cells. The temporal and spatial correlation of these two events suggested that tyrosine phosphorylation of villin may be involved in the rearrangement of the microvillar cytoskeleton. This hypothesis was investigated by analyzing the effects of tyrosine phosphorylation of villin on the kinetics of actin polymerization by reconstituting in vitro the tyrosine phosphorylation of villin and its association with actin. Full-length recombinant human villin was phosphorylated in vitro by expression in the TKX1-competent cells that carry an inducible tyrosine kinase gene. The actin-binding properties of villin were examined using a co-sedimentation assay. Phosphorylation of villin did not change the stoichiometry (1:2) but decreased the binding affinity (4.4 microm for unphosphorylated versus 0.6 microm for phosphorylated) of villin for actin. Using a pyrene-actin-based fluorescence assay, we demonstrated that tyrosine phosphorylation had a negative effect on actin nucleation by villin. In contrast, tyrosine phosphorylation enhanced actin severing by villin. Electron microscopic analysis showed complementary morphological changes. Phosphorylation inhibited the actin bundling and enhanced the actin severing functions of villin. Taken together our data show that tyrosine phosphorylation of villin decreases the amount of villin bound to actin filaments, inhibits the actin-polymerizing properties of villin, and promotes the actin-depolymerizing functions instead. These observations suggest a role for tyrosine phosphorylation in modulating the microvillar cytoskeleton in vivo by villin in response to specific physiological stimuli. PMID- 11500486 TI - Cloning and verification of the Lactococcus lactis pyrG gene and characterization of the gene product, CTP synthase. AB - The pyrG gene of Lactococcus lactis subsp. cremoris, encoding CTP synthase, has been cloned and sequenced. It is flanked upstream by an open reading frame showing homology to several aminotransferases and downstream by an open reading frame of unknown function. L. lactis strains harboring disrupted pyrG alleles were constructed. These mutants required cytidine for growth, proving that in L. lactis, the pyrG product is the only enzyme responsible for the amination of UTP to CTP. In contrast to the situation in Escherichia coli, an L. lactis pyrG mutant could be constructed in the presence of a functional cdd gene encoding cytidine deaminase. A characterization of the enzyme revealed similar properties as found for CTP synthases from other organisms. However, unlike the majority of CTP synthases the lactococcal enzyme can convert dUTP to dCTP, although a half saturation concentration of 0.6 mm for dUTP makes it unlikely that this reaction plays a significant physiological role. As for other CTP synthases, the oligomeric structure of the lactococcal enzyme was found to be a tetramer, but unlike most of the other previously characterized enzymes, the tetramer was very stable even at dilute enzyme concentrations. PMID- 11500487 TI - Phosphoglucomutase is an in vivo lithium target in yeast. AB - Lithium is a drug frequently used in the treatment of manic depressive disorder. We have observed that the yeast Saccharomyces cerevisiae is very sensitive to lithium when growing in galactose medium. In this work we show that lithium inhibits with high affinity yeast (IC50 approximately 0.2 mm) and human (IC50 approximately 1.5 mm) phosphoglucomutase, the enzyme that catalyzes the reversible conversion of glucose 1-phosphate to glucose 6-phosphate. Lithium inhibits the rate of fermentation when yeast are grown in galactose and induces accumulation of glucose 1-phosphate and galactose 1-phosphate. Accumulation of these metabolites was also observed when a strain deleted of the two isoforms of phosphoglucomutase was incubated in galactose medium. In glucose-grown cells lithium reduces the steady state levels of UDP-glucose, resulting in a defect on trehalose and glycogen biosynthesis. Lithium acts as a competitive inhibitor of yeast phosphoglucomutase activity by competing with magnesium, a cofactor of the enzyme. High magnesium concentrations revert lithium inhibition of growth and phosphoglucomutase activity. Lithium stress causes an increase of the phosphoglucomutase activity due to an induction of transcription of the PGM2 gene, and its overexpression confers lithium tolerance in galactose medium. These results show that phosphoglucomutase is an important in vivo lithium target. PMID- 11500488 TI - MMP inhibitors augment fibroblast adhesion through stabilization of focal adhesion contacts and up-regulation of cadherin function. AB - Increased pericellular proteolysis due to an imbalance between MMPs (matrix metalloproteinases) and TIMPs (tissue inhibitors of metalloproteinases) promotes early stages of tumorigenesis. We have reported that TIMP-1 down-regulation confers tumorigenicity on immortal Swiss 3T3 fibroblasts. In pursuit of the mechanism involved in this transformation, we asked whether MMP inhibitors modulate contact inhibition and cell adhesion, because the dysregulation of these events is essential for cellular transformation. Using both genetic and biochemical means, we demonstrate that MMP inhibitors regulate fibroblast cell adhesion. TIMP-1 down-regulated cells formed dense, multilayered colonies, suggesting a loss of contact inhibition. Recombinant TIMP-1 and synthetic MMP inhibitors (MMPi) restored normal cell contact and density of these cells in a dose-dependent manner. Consequently, the effect of MMPi on both cell extracellular matrix (ECM) and cell-cell adhesion were investigated. Upon MMPi treatment, p125(FAK) was redistributed, together with vinculin, to points of cell ECM contact. Furthermore, phosphorylation of p125(FAK) was restored to levels similar to that of wild type. In parallel, MMPi treatment increased cadherin levels and stabilized cadherin-mediated cell-cell contacts. Moreover, enhanced cadherin function was evident as increased calcium-dependent cell-cell aggregation and co-localization of cadherin and beta-catenin at the cell membrane. We also obtained independent evidence of altered cadherin function using timp-1(-/-) mouse embryonic fibroblasts. Our data provide provocative evidence that increased pericellular proteolysis impacts cell adhesion systems to offset normal contact inhibition, with subsequent effects on cell transformation and tumorigenesis. PMID- 11500489 TI - Cysteine 38 in p65/NF-kappaB plays a crucial role in DNA binding inhibition by sesquiterpene lactones. AB - Sesquiterpene lactones (SLs) have potent anti-inflammatory properties. We have shown previously that they exert this effect in part by inhibiting activation of the transcription factor NF-kappaB, a central regulator of the immune response. We have proposed a molecular mechanism for this inhibition based on computer molecular modeling data. In this model, SLs directly alkylate the p65 subunit of NF-kappaB, thereby inhibiting DNA binding. Nevertheless, an experimental evidence for the proposed mechanism was lacking. Moreover, based on experiments using the SL parthenolide, an alternative mode of action has been proposed by other authors in which SLs inhibit IkappaB-alpha degradation. Here we report the construction of p65/NF-kappaB point mutants that lack the cysteine residues alkylated by SLs in our model. In contrast to wild type p65, DNA-binding of the Cys(38) --> Ser and Cys(38,120) --> Ser mutants is no longer inhibited by SLs. In addition, we provide evidence that parthenolide uses a similar mechanism to other SLs in inhibiting NF-kappaB. Contrary to previous reports, we show that parthenolide, like other SLs, inhibits NF-kappaB most probably by alkylating p65 at Cys(38). Although a slight inhibition of IkappaB degradation was detected for all SLs, the amount of remaining IkappaB was too low to explain the observed NF-kappaB inhibition. PMID- 11500490 TI - ERK2- and p90(Rsk2)-dependent pathways regulate the CCAAT/enhancer-binding protein-beta interaction with serum response factor. AB - The serum response element (SRE) of the c-fos promoter is a convergence point for mitogenic signaling pathways. Several transcription factors regulate SRE, including serum response factor (SRF), ternary complex factors, and CCAAT/enhancer-binding protein-beta (C/EBPbeta). C/EBPbeta can interact with both SRF and the ternary complex factor family member Elk-1, but only in response to activated Ras. Transactivation of the SRE by C/EBPbeta is also greatly stimulated by Ras. The Ras effectors that signal to C/EBPbeta are unknown. In this report, we demonstrate that a consensus MAPK site in C/EBPbeta is necessary for Ras stimulation of both C/EBPbeta-SRF interaction and transactivation of the SRE by C/EBPbeta. To dissect signaling pathways activated downstream of Ras, different Ras effector constructs were analyzed. We show that activated forms of Raf and phosphatidylinositol 3-kinase stimulate C/EBPbeta-SRF interaction. We also show a novel selectivity for the MAPK family member ERK2, where dominant-negative ERK2, but not dominant-negative ERK1, blocks Ras stimulation of C/EBPbeta-SRF interaction. In addition, recombinant C/EBPbeta protein is phosphorylated by ERK2, but not by ERK1, in vitro. Finally, we demonstrate a requirement for p90(Rsk2) in regulation of C/EBPbeta-SRF interaction. These data show that multiple Ras effectors are required to regulate C/EBPbeta and SRF association. PMID- 11500491 TI - Calcium entry through L-type calcium channels causes mitochondrial disruption and chromaffin cell death. AB - Sustained, mild K+ depolarization caused bovine chromaffin cell death through a Ca(2+)-dependent mechanism. During depolarization, Ca(2+) entered preferentially through L-channels to induce necrotic or apoptotic cell death, depending on the duration of the cytosolic Ca(2+) concentration ([Ca(2+)](c)) signal, as proven by the following. (i) The L-type Ca(2+) channel activators Bay K 8644 and FPL64176, more than doubled the cytotoxic effects of 30 mm K+; (ii) the L-type Ca(2+) channel blocker nimodipine suppressed the cytotoxic effects of K+ alone or K+ plus FPL64176; (iii) the potentiation by FPL64176 of the K+ -evoked [Ca(2+)](c) elevation was totally suppressed by nimodipine. Cell exposure to K+ plus the L type calcium channel agonist FPL64176 caused an initial peak rise followed by a sustained elevation of the [Ca(2+)](c) that, in turn, increased [Ca(2+)](m) and caused mitochondrial membrane depolarization. Cyclosporin A, a blocker of the mitochondrial transition pore, and superoxide dismutase prevented the apoptotic cell death induced by Ca(2+) overload through L-channels. These results suggest that Ca(2+) entry through L-channels causes both calcium overload and mitochondrial disruption that will lead to the release of mediators responsible for the activation of the apoptotic cascade and cell death. This predominant role of L-type Ca(2+) channels is not shared by other subtypes of high threshold voltage-dependent neuronal Ca(2+) channels (i.e. N, P/Q) expressed by bovine chromaffin cells. PMID- 11500492 TI - Alanine scanning mutagenesis of a type 1 insulin-like growth factor receptor ligand binding site. AB - The high resolution crystal structure of an N-terminal fragment of the IGF-I receptor, has been reported. While this fragment is itself devoid of ligand binding activity, mutational analysis has indicated that its N terminus (L1, amino acids 1-150) and the C terminus of its cysteine-rich domain (amino acids 190-300) contain ligand binding determinants. Mutational analysis also suggests that amino acids 692-702 from the C terminus of the alpha subunit are critical for ligand binding. A fusion protein, formed from these fragments, binds IGF-I with an affinity similar to that of the whole extracellular domain, suggesting that these are the minimal structural elements of the IGF-I binding site. To further characterize the binding site, we have performed structure directed and alanine-scanning mutagenesis of L1, the cysteine-rich domain and amino acids 692 702. Alanine mutants of residues in these regions were transiently expressed as secreted recombinant receptors and their affinity was determined. In L1 alanine mutants of Asp(8), Asn(11), Tyr(28), His(30), Leu(33), Leu(56), Phe(58), Arg(59), and Trp(79) produced a 2- to 10-fold decrease in affinity and alanine mutation of Phe(90) resulted in a 23-fold decrease in affinity. In the cysteine-rich domain, mutation of Arg(240), Phe(241), Glu(242), and Phe(251) produced a 2- to 10-fold decrease in affinity. In the region between amino acids 692 and 702, alanine mutation of Phe(701) produced a receptor devoid of binding activity and alanine mutations of Phe(693), Glu(693), Asn(694), Leu(696), His(697), Asn(698), and Ile(700) exhibited decreases in affinity ranging from 10- to 30-fold. With the exception of Trp(79), the disruptive mutants in L1 form a discrete epitope on the surface of the receptor. Those in the cysteine-rich domain essential for intact affinity also form a discrete epitope together with Trp(79). PMID- 11500493 TI - The Npr1 kinase controls biosynthetic and endocytic sorting of the yeast Gap1 permease. AB - Membrane trafficking of the general amino acid permease (Gap1) of Saccharomyces cerevisiae is under nitrogen regulation. In cells growing on proline or urea as the sole nitrogen source, newly synthesized Gap1 is delivered to the plasma membrane, where it accumulates. Upon addition of NH(4)(+), a preferential nitrogen source, Gap1 is endocytosed and targeted to the vacuole, where it is degraded. This down-regulation requires ubiquitination of the permease, and this ubiquitination is dependent on the essential Npi1/Rsp5 ubiquitin ligase. In this study, we investigated the role of the Npr1 kinase in the regulation of Gap1 trafficking. We show that Npr1 is required for stabilization of Gap1 at the plasma membrane: when an npr1(ts) mutant growing on proline is shifted to the restrictive temperature, Gap1 down-regulation is triggered, as it is when NH(4)(+) is added to wild-type cells. The fate of newly synthesized Gap1 en route to the plasma membrane is also under Npr1 control: in an npr1Delta mutant, neosynthesized Gap1 is sorted from the Golgi to the vacuole without passing via the plasma membrane. Similar direct sorting of neosynthesized Gap1 to the vacuole was observed in wild-type cells grown on NH(4)(+). Finally, Gap1 is phosphorylated in NPR1 cells, but this phosphorylation is not strictly dependent on Npr1. Our results show that Npr1 kinase plays a central role in the physiological control of Gap1 trafficking and that this control is exerted not only on Gap1 present at the plasma membrane but also on Gap1 late in the secretory pathway. Npr1 belongs to a subgroup of protein kinases, some of which are reported to exert a positive control on the activity of other permeases. We propose that these kinases also function as regulators of permease trafficking. PMID- 11500494 TI - Ubiquitin is required for sorting to the vacuole of the yeast general amino acid permease, Gap1. AB - In yeast, ubiquitin plays a central role in proteolysis of a multitude of proteins and serves also as a signal for endocytosis of many plasma membrane proteins. We showed previously that ubiquitination of the general amino acid permease (Gap1) is essential to its endocytosis followed by vacuolar degradation. These processes occur when NH(4)(+), a preferential source of nitrogen, is added to cells growing on proline or urea, i.e. less favored nitrogen sources. In this study, we show that Gap1 is ubiquitinated on two lysine residues in the cytosolic N terminus (positions 9 and 16). A mutant Gap1 in which both lysines are mutated (Gap1(K9K16)) remains fully stable at the plasma membrane after NH(4)(+) addition. Furthermore, each of the two lysines harbors a poly-ubiquitin chain in which ubiquitin is linked to the lysine 63 of the preceding ubiquitin. The Gap1(K9) and Gap1(K16) mutants, in which a single lysine is mutated, are down regulated in response to NH(4)(+) although more slowly. In proline-grown cells lacking Npr1, a protein kinase involved in the control of Gap1 trafficking, newly synthesized Gap1 is sorted from the Golgi to the vacuole without passing through the plasma membrane (accompanying article, De Craene, J.-O., Soetens, O., and Andre, B. (2001) J. Biol. Chem. 276, 43939-43948). We show here that ubiquitination of Gap1 is also required for this direct sorting to the vacuole. In an npr1Delta mutant, neosynthesized Gap1(K9K16) is rerouted to and accumulates at the plasma membrane. Finally, Bul1 and Bul2, two proteins interacting with Npi1/Rsp5, are essential to ubiquitination and down-regulation of cell-surface Gap1, as well as to sorting of neosynthesized Gap1 to the vacuole, as occurs in an npr1Delta mutant. Our results reveal a novel role of ubiquitin in the control of Gap1 trafficking, i.e. direct sorting from the late secretory pathway to the vacuole. This result reinforces the growing evidence that ubiquitin plays an important role not only in internalization of plasma membrane proteins but also in their sorting in the endosomes and/or trans-Golgi. PMID- 11500495 TI - Effects of the membrane dipole potential on the interaction of saquinavir with phospholipid membranes and plasma membrane receptors of Caco-2 cells. AB - The combined use of the membrane surface potential fluorescent sensor fluorescein phosphatidylethanolamine (FPE) and the membrane dipole potential fluorescent sensor di-8-ANEPPS to characterize the interaction of molecules with model and cellular membranes and to asses the influence of the dipole potential on the interaction is reported. The study of the human immunodeficiency virus protease inhibitor saquinavir with Caco-2 cells and phospholipid membranes reveals that the compound interacts with the lipidic bilayer of model membranes with a simple hyperbolic binding profile but with Caco-2 cells in a cooperative way involving membrane receptors. Additional studies indicated that colchicine acts as a competitor ligand to saquinavir and suggests, in agreement with other reports, that the identity of the saquinavir "receptor" could be P-glycoprotein or the multiple drug resistance-associated protein. The modification of the magnitude of the membrane dipole potential using compounds such as cholesterol, phloretin, and 6-ketocholestanol influences the binding capacity of saquinavir. Furthermore, removal of cholesterol from the cell membrane using methyl-beta-cyclodextrin significantly decreases the binding capacity of saquinavir. Because removal of cholesterol from the cell membrane has been reported to disrupt membrane domains known as "rafts," our observations imply that the membrane dipole potential plays an important role as a modulator of molecule-membrane interactions in these membrane structures. Such a role is suggested to contribute to the altered behavior of receptor-mediated signaling systems in membrane rafts. PMID- 11500496 TI - MRG15 activates the B-myb promoter through formation of a nuclear complex with the retinoblastoma protein and the novel protein PAM14. AB - The MORF4-Related Gene on chromosome 15 (MRG15) is a member of a novel family of genes originally identified in studies to reveal cell senescence-inducing factors. MRG15 contains several predicted protein motifs, including a nuclear localization signal, a helix-loop-helix region, a leucine zipper, and a chromodomain. These motifs are commonly associated with transcription factors, suggesting that MRG15 may likewise function as a transcriptional regulator. To examine the potential function(s) of MRG15, we sought to identify cellular factors associated with this MRG family member. In this regard, we have found that both the retinoblastoma tumor suppressor (Rb) and a novel nuclear protein PAM14 (Protein Associated with MRG, 14 kDa) specifically associate with MRG15. We have further demonstrated that these interactions require the helix-loop-helix and leucine zipper domains of MRG15. Interestingly we have found all three proteins present in a multiprotein complex, suggesting that at least some of their functions may be interdependent. Although the functions of PAM14 have yet to be elucidated, Rb has several well characterized activities, including repression of E2F-activated promoters such as that of B-myb. Significantly we have demonstrated that MRG15 blocks the Rb-induced repression of this promoter, leading to B-myb promoter activation. Collectively these results suggest that MRG15 regulates transcription through interactions with a cellular protein complex containing Rb and PAM14. PMID- 11500497 TI - Cloning and characterization of mouse UBPy, a deubiquitinating enzyme that interacts with the ras guanine nucleotide exchange factor CDC25(Mm)/Ras-GRF1. AB - We used yeast "two-hybrid" screening to isolate cDNA-encoding proteins interacting with the N-terminal domain of the Ras nucleotide exchange factor CDC25(Mm). Three independent overlapping clones were isolated from a mouse embryo cDNA library. The full-length cDNA was cloned by RACE-polymerase chain reaction. It encodes a large protein (1080 amino acids) highly homologous to the human deubiquitinating enzyme hUBPy and contains a well conserved domain typical of ubiquitin isopeptidases. Therefore we called this new protein mouse UBPy (mUBPy). Northern blot analysis revealed a 4-kilobase mRNA present in several mouse tissues and highly expressed in testis; a good level of expression was also found in brain, where CDC25(Mm) is exclusively expressed. Using a glutathione S transferase fusion protein, we demonstrated an "in vitro" interaction between mUBPy and the N-terminal half (amino acids 1-625) of CDC25(Mm). In addition "in vivo" interaction was demonstrated after cotransfection in mammalian cells. We also showed that CDC25(Mm), expressed in HEK293 cells, is ubiquitinated and that the coexpression of mUBPy decreases its ubiquitination. In addition the half-life of CDC25Mm protein was considerably increased in the presence of mUBPy. The specific function of the human homolog hUBPy is not defined, although its expression was correlated with cell proliferation. Our results suggest that mUBPy may play a role in controlling degradation of CDC25(Mm), thus regulating the level of this Ras-guanine nucleotide exchange factor. PMID- 11500498 TI - Effect of insulin on cell cycle progression in MCF-7 breast cancer cells. Direct and potentiating influence. AB - We recently demonstrated that in MCF-7 breast cancer cells, insulin promoted the phosphorylation and activation of geranylgeranyltransferase I (GGTI-I), increased the amounts of geranylgeranylated Rho-A and potentiated the transactivating activity of lysophosphatidic acid (LPA) (Chappell, J., Golovchenko, I., Wall, K., Stjernholm, R., Leitner, J., Goalstone, M., and Draznin, B. (2000) J. Biol. Chem. 275, 31792-31797). In the present study, we explored the mechanism of this potentiating effect of insulin on LPA. Insulin (10 nm) potentiated the ability of LPA to stimulate cell cycle progression and DNA synthesis in MCF-7 cells. The potentiating effect of insulin appears to involve increases in the expression of cyclin E and decreases in the expression of the cyclin-dependent kinase inhibitor p27Kip1. All potentiating effects of insulin were inhibited in the presence of an inhibitor of GGTase I, GGTI-286 (3 microm) or by an expression of a dominant negative mutant of Rho-A. In contrast to its potentiating action, a direct mitogenic effect of insulin in MCF-7 cells involves activation of phosphatidylinositol 3-kinase and increased expression of cyclin D1. We conclude that the ability of insulin to increase the cellular amounts of geranylgeranylated Rho-A results in potentiation of the LPA effect on cyclin E expression and degradation of p27Kip1 and cell cycle progression in MCF-7 breast cancer cells. PMID- 11500499 TI - Prenylation of target GTPases contributes to signaling specificity of Ras-guanine nucleotide exchange factors. AB - Ras-GRF1 and Ras-GRF2 constitute a family of calmodulin-regulated guanine nucleotide exchange factors (GEFs) that activate Ras proteins. Here we show that whereas Ras-GRF1 activated both Ha-Ras and R-Ras in cells, Ras-GRF2 activated only Ha-Ras. The inability of Ras-GRF2 to activate R-Ras was the consequence of the GTPase being post-translationally modified, since Ras-GRF2 activated unprocessed R-Ras as effectively as unprocessed Ha-Ras when assays were performed either in vivo or in vitro. Moreover, Ras-GRF2 failed to activate fully processed R-Ras in vitro. The particular C-terminal lipid attached to the GTPases played an important role in determining signaling specificity, since R-Ras became more responsive to Ras-GRF2 when it was farnesylated instead of geranylgeranylated. Similarly, Ha-Ras became less responsive to Ras-GRF2 when it was geranylgeranylated instead of farnesylated. Analysis of chimeras between Ras-GRF1 and Ras-GRF2 demonstrated that a 30-amino acid segment embedded with their catalytic domains was responsible for recognizing the presence of different lipids on Ras proteins. These results indicate that the specific lipid moiety attached to GTPases can contribute to signaling specificity of Ras-GEFs. PMID- 11500500 TI - New insights into the heparan sulfate proteoglycan-binding activity of apolipoprotein E. AB - Defective binding of apolipoprotein E (apoE) to heparan sulfate proteoglycans (HSPGs) is associated with increased risk of atherosclerosis due to inefficient clearance of lipoprotein remnants by the liver. The interaction of apoE with HSPGs has also been implicated in the pathogenesis of Alzheimer's disease and may play a role in neuronal repair. To identify which residues in the heparin-binding site of apoE and which structural elements of heparan sulfate interact, we used a variety of approaches, including glycosaminoglycan specificity assays, (13)C nuclear magnetic resonance, and heparin affinity chromatography. The formation of the high affinity complex required Arg-142, Lys-143, Arg-145, Lys-146, and Arg 147 from apoE and N- and 6-O-sulfo groups of the glucosamine units from the heparin fragment. As shown by molecular modeling, using a high affinity binding octasaccharide fragment of heparin, these findings are consistent with a binding mode in which five saccharide residues of fully sulfated heparan sulfate lie in a shallow groove of the alpha-helix that contains the HSPG-binding site (helix 4 of the four-helix bundle of the 22-kDa fragment). This groove is lined with residues Arg-136, Ser-139, His-140, Arg-142, Lys-143, Arg-145, Lys-146, and Arg-147. In the model, all of these residues make direct contact with either the 2-O-sulfo groups of the iduronic acid monosaccharides or the N- and 6-O-sulfo groups of the glucosamine sulfate monosaccharides. This model indicates that apoE has an HSPG binding site highly complementary to heparan sulfate rich in N- and O-sulfo groups such as that found in the liver and the brain. PMID- 11500501 TI - Fucosylation of Cripto is required for its ability to facilitate nodal signaling. AB - O-linked fucose modification is rare and has been shown to occur almost exclusively within epidermal growth factor (EGF)-like modules. We have found that the EGF-CFC family member human Cripto-1 (CR) is modified with fucose and through a combination of peptide mapping, mass spectrometry, and sequence analysis localized the site of attachment to Thr-88. The identification of a fucose modification on human CR within its EGF-like domain and the presence of a consensus fucosylation site within all EGF-CFC family members suggest that this is a biologically important modification in CR, which functionally distinguishes it from the EGF ligands that bind the type 1 erbB growth factor receptors. A single CR point mutation, Thr-88 --> Ala, results in a form of the protein that is not fucosylated and has substantially weaker activity in cell-based CR/Nodal signaling assays, indicating that fucosylation is functionally important for CR to facilitate Nodal signaling. PMID- 11500502 TI - Characterization of the binding interface between the copper chaperone Atx1 and the first cytosolic domain of Ccc2 ATPase. AB - The interaction of the copper chaperone Atx1 and the first cytosolic domain of Ccc2 ATPase, Ccc2a, was investigated by NMR in solution. In particular, a solution of Cu(I)-15NAtx1 was titrated with apo-Ccc2a, and, vice versa, a solution of Cu(I)-15NCcc2a was titrated with apo-Atx1. By following the 15N and 1H chemical shifts, a new species is detected in both experiments. This species is the same in both titrations and is in fast exchange with the parent species on the NMR time scale. Nuclear relaxation data are consistent with the formation of an adduct. Judging from the nuclear Overhauser effect spectroscopy patterns, the structure of Cu(I)-15NCcc2a in the presence of apo-Atx1 is not significantly altered, whereas Cu(I)-15NAtx1 in the presence of apo-Ccc2a experiences some changes with respect to both the apoproteins and the Cu(I)-loaded proteins. The structure of the Cu(I)-15NAtx1 moiety in the adduct was obtained from 1137 nuclear Overhauser effects to a final root mean square deviation to the mean structure of 0.76 +/- 0.13 A for the backbone and 1.11 +/- 0.11 A for the heavy atoms. 15N and 1H chemical shifts suggest the regions of interaction that, together with independent information, allow a structural model of the adduct to be proposed. The apo form of Atx1 displays significant mobility in loops 1 and 5, the N-terminal part of helix alpha1, and the C-terminal part of helix alpha2 on the ms-micros time scale. These regions correspond to the metal binding site. Such mobility is largely reduced in the free Cu(I)-Atx1 and in the adduct with apo-Ccc2a. The analogous mobility of Ccc2a in both Cu(I) and apo forms is reduced with respect to Atx1. Such an adduct is relevant as a structural and kinetic model for copper transfer from Atx1 to Ccc2a in physiological conditions. PMID- 11500503 TI - Differential dependence of the D1 and D5 dopamine receptors on the G protein gamma 7 subunit for activation of adenylylcyclase. AB - The D(1) dopamine receptor, G protein gamma(7) subunit, and adenylylcyclase are selectively expressed in the striatum, suggesting their potential interaction in a common signaling pathway. To evaluate this possibility, a ribozyme strategy was used to suppress the expression of the G protein gamma(7) subunit in HEK 293 cells stably expressing the human D(1) dopamine receptor. Prior in vitro analysis revealed that the gamma(7) ribozyme possessed cleavage activity directed exclusively toward the gamma(7) RNA transcript (Wang, Q., Mullah, B., Hansen, C., Asundi, J., and Robishaw, J. D. (1997) J. Biol. Chem. 272, 26040-26048). In vivo analysis of cells transfected with the gamma(7) ribozyme showed a specific reduction in the expression of the gamma(7) protein. Coincident with the loss of the gamma(7) protein, there was a noticeable reduction in the expression of the beta(1) protein, confirming their interaction in these cells. Finally, functional analysis of ribozyme-mediated suppression of the beta(1) and gamma(7) proteins revealed a significant attenuation of SKF81297-stimulated adenylylcyclase activity in D(1) dopamine receptor-expressing cells. By contrast, ribozyme mediated suppression of the beta(1) and gamma(7) proteins showed no reduction of SKF81297-stimulated adenylylcyclase activity in D(5) dopamine receptor-expressing cells. Taken together, these data indicate that the structurally related D(1) and D(5) dopamine receptor subtypes utilize G proteins composed of distinct betagamma subunits to stimulate adenylylcyclase in HEK 293 cells. Underscoring the physiological relevance of these findings, single cell reverse transcriptase polymerase chain reaction analysis revealed that the D(1) dopamine receptor and the G protein gamma(7) subunit are coordinately expressed in substance P containing neurons in rat striatum, suggesting that the G protein gamma(7) subunit may be a new target for drugs to selectively alter dopaminergic signaling within the brain. PMID- 11500504 TI - Insulin-like growth factor-mediated muscle differentiation: collaboration between phosphatidylinositol 3-kinase-Akt-signaling pathways and myogenin. AB - The differentiation and maturation of skeletal muscle require interactions between signaling pathways activated by hormones and growth factors and an intrinsic regulatory network controlled by myogenic transcription factors. Insulin-like growth factors (IGFs) play key roles in muscle development in the embryo and in regeneration in the adult. To study mechanisms of IGF action in muscle, we developed a myogenic cell line that overexpresses IGF-binding protein 5. C2BP5 cells remain quiescent in low serum differentiation medium until the addition of IGF-I. Here we use this cell line to identify signaling pathways controlling IGF-mediated differentiation. Induction of myogenin by IGF-I and myotube formation were prevented by the phosphatidylinositol (PI) 3-kinase inhibitor, LY294002, even when included 2 days after growth factor addition, whereas expression of active PI 3-kinase could promote differentiation in the absence of IGF-I. Differentiation also was induced by myogenin but was blocked by LY294002. The differentiation-promoting effects of IGF-I were mimicked by a modified membrane-targeted inducible Akt-1 (iAkt), and iAkt was able to stimulate differentiation of C2 myoblasts and primary mouse myoblasts incubated with otherwise inhibitory concentrations of LY294002. These results show that an IGF regulated PI 3-kinase-Akt pathway controls muscle differentiation by mechanisms acting both upstream and downstream of myogenin. PMID- 11500505 TI - Mutation of Trp1254 in the multispecific organic anion transporter, multidrug resistance protein 2 (MRP2) (ABCC2), alters substrate specificity and results in loss of methotrexate transport activity. AB - The ATP-binding cassette (ABC) proteins comprise a large superfamily of transmembrane transporters that utilize the energy of ATP hydrolysis to translocate their substrates across biological membranes. Multidrug resistance protein (MRP) 2 (ABCC2) belongs to subfamily C of the ABC superfamily and, when overexpressed in tumor cells, confers resistance to a wide variety of anticancer chemotherapeutic agents. MRP2 is also an active transporter of organic anions such as methotrexate (MTX), estradiol glucuronide (E217betaG), and leukotriene C4 and is located on the apical membrane of polarized cells including hepatocytes where it acts as a biliary transporter. We recently identified a highly conserved tryptophan residue in the related MRP1 that is critical for the substrate specificity of this protein. In the present study, we have examined the effect of replacing the analogous tryptophan residue at position 1254 of MRP2. We found that only nonconservative substitutions (Ala and Cys) of Trp1254 eliminated [3H]E217betaG transport by MRP2, whereas more conservative substitutions (Phe and Tyr) had no effect. In addition, only the most conservatively substituted mutant (W1254Y) transported [3H]leukotriene C4, whereas all other substitutions eliminated transport of this substrate. On the other hand, all substitutions of Trp1254 eliminated transport of [3H]MTX. Finally, we found that sulfinpyrazone stimulated [3H]E217betaG transport by wild-type MRP2 4-fold, whereas transport by the Trp1254 substituted mutants was enhanced 6-10-fold. In contrast, sulfinpyrazone failed to stimulate [3H]MTX transport by either wild-type MRP2 or the MRP2-Trp1254 mutants. Taken together, our results demonstrate that Trp1254 plays an important role in the ability of MRP2 to transport conjugated organic anions and identify this amino acid in the putative last transmembrane segment (TM17) of this ABC protein as being critical for transport of MTX. PMID- 11500506 TI - Galpha(i2) enhances insulin signaling via suppression of protein-tyrosine phosphatase 1B. AB - Suppression of the expression of the heterotrimeric G-protein Galpha(i2) in vivo has been shown to provoke insulin resistance, whereas enhanced insulin signaling is observed when Galpha(i2) is overexpressed in vivo. The basis for Galpha(i2) regulation of insulin signaling was explored in transgenic mice with targeted expression of the GTPase-deficient, constitutively active Q205L Galpha(i2) in fat and skeletal muscle. Phosphorylation of insulin receptor and IRS-1 in response to insulin challenge in vivo was markedly amplified in fat and skeletal muscle expressing Q205L Galpha(i2). The expression and activity of the protein-tyrosine phosphatase 1B (PTP1B), but not protein-tyrosine phosphatases SHP-1, SHP-2, and LAR, were constitutively decreased in tissues expressing the Q205L Galpha(i2), providing a direct linkage between insulin signaling and Galpha(i2). The loss of PTP1B expression may explain, in part, the loss of PTP1B activity in the iQ205L transgenic mice. Activation of Galpha(i2) in mouse adipocytes with lysophosphatidic acid was shown to decrease PTP1B activity, whereas pertussis toxin inactivates Galpha(i2), blocks lysophosphatidic acid-stimulated inhibition of PTP1B activity, and blocks tonic suppression of PTP1B activity by Galpha(i2). Elevation of intracellular cAMP in fat cells is shown to increase PTP1B activity, whereas either depression of cAMP levels or direct activation of Galpha(i2) suppresses PTP1B. These data provide the first molecular basis for the interplay between Galpha(i2) and insulin signaling, i.e. activation of Galpha(i2) can suppress both the expression and activity of PTP1B in insulin-sensitive tissues. PMID- 11500507 TI - MD-2 binds to bacterial lipopolysaccharide. AB - The exact roles and abilities of the individual components of the lipopolysaccharide (LPS) receptor complex of proteins remain unclear. MD-2 is a molecule found in association with toll-like receptor 4. We produced recombinant human MD-2 to explore its LPS binding ability and role in the LPS receptor complex. MD-2 binds to highly purified rough LPS derived from Salmonella minnesota and Escherichia coli in five different assays; one assay yielded an apparent KD of 65 nm. MD-2 binding to LPS did not require LPS-binding proteins LBP and CD14; in fact LBP competed with MD-2 for LPS. MD-2 enhanced the biological activity of LPS in toll-like receptor 4-transfected Chinese hamster ovary cells but inhibited LPS activation of U373 astrocytoma cells and of monocytes in human whole blood. These data indicate that MD-2 is a genuine LPS binding protein and strongly suggest that MD-2 could play a role in regulation of cellular activation by LPS depending on its local availability. PMID- 11500508 TI - A fraction of yeast Cu,Zn-superoxide dismutase and its metallochaperone, CCS, localize to the intermembrane space of mitochondria. A physiological role for SOD1 in guarding against mitochondrial oxidative damage. AB - Cu,Zn-superoxide dismutase (SOD1) is an abundant, largely cytosolic enzyme that scavenges superoxide anions. The biological role of SOD1 is somewhat controversial because superoxide is thought to arise largely from the mitochondria where a second SOD (manganese SOD) already resides. Using bakers' yeast as a model, we demonstrate that Cu,Zn-SOD1 helps protect mitochondria from oxidative damage, as sod1Delta mutants show elevated protein carbonyls in this organelle. In accordance with this connection to mitochondria, a fraction of active SOD1 localizes within the intermembrane space (IMS) of mitochondria together with its copper chaperone, CCS. Neither CCS nor SOD1 contains typical N terminal presequences for mitochondrial uptake; however, the mitochondrial accumulation of SOD1 is strongly influenced by CCS. When CCS synthesis is repressed, mitochondrial SOD1 is of low abundance, and conversely IMS SOD1 is very high when CCS is largely mitochondrial. The mitochondrial form of SOD1 is indeed protective against oxidative damage because yeast cells enriched for IMS SOD1 exhibit prolonged survival in the stationary phase, an established marker of mitochondrial oxidative stress. Cu,Zn-SOD1 in the mitochondria appears important for reactive oxygen physiology and may have critical implications for SOD1 mutations linked to the fatal neurodegenerative disorder, amyotrophic lateral sclerosis. PMID- 11500510 TI - Multiple levels of control regulate the yeast cAMP-response element-binding protein repressor Sko1p in response to stress. AB - The Sko1p transcriptional repressor regulates a subset of osmoinducible stress defense genes in Saccharomyces cerevisiae by binding to cAMP-responsive elements. We have reported previously that in response to stress Sko1p is phosphorylated by the stress-activated Hog1p mitogen-activated protein kinase, which disrupts its interaction with the Ssn6p x Tup1p corepressor. Here we report that other mechanisms are essential for the regulation of the Sko1p repressor activity upon stress. The nuclear localization of Sko1p depends on the stress-inhibited protein kinase A (PKA). Sko1p is localized in the nucleus of unstressed cells, and it redistributes to the cytosol upon severe salt stress (1 m NaCl). Yeast mutants with low PKA activity localize Sko1p to the cytoplasm in the absence of stress and exhibit deregulated expression of cAMP-responsive element-regulated genes. The central part (315) of Sko1p, containing the PKA phosphorylation sites and the basic domain-leucine zipper domain, is essential for its nuclear localization. Salt-induced export of Sko1p from the nucleus is independent of Hog1p and of the Bcy1p regulatory subunit of PKA. Furthermore, phosphorylation by PKA slightly enhanced DNA binding affinity of Sko1p in vitro, whereas Sko1p dimerization in vivo is not regulated by stress. Sko1p repressor activity is associated to its binding to the Ssn6p x Tup1p complex. Interestingly, the Sko1p NH(2) terminus (1), containing the Hog1p phosphorylation sites, associates in vivo with Tup1p in the absence of Ssn6p, suggesting that Sko1p represses gene transcription by interacting directly with the Tup1p subunit of the Ssn6p x Tup1p complex. PMID- 11500509 TI - Suppression of tumor necrosis factor-mediated apoptosis by nuclear factor kappaB independent bone morphogenetic protein/Smad signaling. AB - The activation of nuclear factor kappaB (NF-kappa B) plays a pivotal role in the regulation of tumor necrosis factor (TNF)-mediated apoptosis. However, little is known about the regulation of TNF-mediated apoptosis by other signaling pathways or growth factors. Here, unexpectedly, we found that bone morphogenetic protein (BMP)-2 and BMP-4 inhibited TNF-mediated apoptosis by inhibition of caspase-8 activation in C2C12 cells, a pluripotent mesenchymal cell line that has the potential to differentiate into osteoblasts depending on BMP stimulation. Utilizing both a trans-dominant IkappaBalpha inhibitor of NF-kappaB expressed in C2C12 cells and IkappaB kinase beta-deficient embryonic mouse fibroblast, we show that BMP-mediated survival was independent of NF-kappaB activation. Rather, the antiapoptotic activity of BMPs functioned through the Smad signaling pathway. Thus, these findings provide the first report of a BMP/Smad signaling pathway that can inhibit TNF-mediated apoptosis, independent of the prosurvival activity of NF-kappaB. Our results suggest that BMPs not only stimulate osteoblast differentiation but can also promote cell survival during the induction of bone formation, offering new insight into the biological functions of BMPs. PMID- 11500511 TI - The fate of desmosomal proteins in apoptotic cells. AB - Activation of caspases results in the disruption of structural and signaling networks in apoptotic cells. Recent biochemical and cell biological studies have shown that components of the cadherin-catenin adhesion complex in epithelial adherens junctions are targeted by caspases during apoptosis. In epithelial cells, desmosomes represent a second type of anchoring junctions mediating strong cell-cell contacts. Using antibodies directed against a set of desmosomal proteins, we show that desmosomes are proteolytically targeted during apoptosis. Desmogleins and desmocollins, representing desmosome-specific members of the cadherin superfamily of cell adhesion molecules, are specifically cleaved after onset of apoptosis. Similar to E-cadherin, the desmoglein-3 cytoplasmic tail is cleaved by caspases. In addition the extracellular domains of desmoglein-3 and desmocollin-3 are released from the cell surface by a metalloproteinase activity. In the presence of caspase and/or metalloproteinase inhibitors, both cleavage reactions are almost completely inhibited. As reported previously, the desmosomal plaque protein plakoglobin is cleaved by caspase-3 during apoptosis. Our studies now show that plakophilin-1 and two other major plaque proteins, desmoplakin-1 and -2, are also cleaved by caspases. Immunofluorescence analysis confirmed that this cleavage results in the disruption of the desmosome structure and thus contributes to cell rounding and disintegration of the intermediate filament system. PMID- 11500512 TI - Characterization of the human ABCG1 gene: liver X receptor activates an internal promoter that produces a novel transcript encoding an alternative form of the protein. AB - The human ABCG1 gene encodes a member of the ATP-binding cassette (ABC) superfamily of transporter proteins and is highly induced when macrophages are incubated with oxysterols. Using mRNA from oxysterol-treated human THP-1 cells together with 5'-rapid amplification of cDNA ends and polymerase chain reaction, we identified a novel ABCG1 transcript that encodes a putative protein of 786 residues containing a new amino terminus of 203 amino acids. Characterization of the genomic organization and structure of the human ABCG1 gene demonstrates that: (i) the gene consists of 23 exons spanning 98 kilobase pairs (kb) on chromosome 21q22.3, (ii) the 203 amino acids are encoded on three previously unidentified exons, 8-10, and (iii) a promoter, containing a TATA box and two liver X receptor (LXR) alpha response elements (LXREs), is located upstream of exon 8. Northern analysis using exon-specific probes confirms that oxysterol treatment results in >10-fold induction of ABCG1 transcripts that are derived from either exons 8-23 or exons 5, 7, and 11-23. Electromobility shift assays demonstrate that LXRalpha and retinoid X receptor alpha bind to the two LXREs in intron 7. Cells were transiently transfected with reporter luciferase constructs under the control of either (i) 9 kb of genomic DNA corresponding to intron 7 and part of exon 8 and containing either wild-type or mutant LXREs or (ii) two copies of the wild-type or mutant LXRE. In all cases, the wild-type construct was regulated in an LXR- and oxysterol-dependent manner, and this regulation was attenuated when the LXREs were mutated. In conclusion, the human ABCG1 gene contains multiple promoters, spans more than 98 kb and comprises 23 exons that give rise to alternative transcripts encoding proteins with different amino-terminal sequences. Elucidation of the various roles of different ABCG1 isoforms will be important for our understanding of mammalian cholesterol homeostasis. PMID- 11500513 TI - The eighth FIII domain of human fibronectin promotes integrin alpha5beta1 binding via stabilization of the ninth FIII domain. AB - Binding of the extracellular matrix molecule fibronectin to the integrin receptor alpha(5)beta(1) elicits downstream signaling pathways that modulate cell function. Fibronectin-alpha(5)beta(1) interaction occurs via the conserved RGD sequence in the tenth FIII (FIII10) domain of fibronectin. A synergistic site containing the sequence PHSRN in the adjacent FIII9 domain has also been identified. Here we investigate the function of the eighth FIII domain in integrin-mediated cell adhesion using a wide range of methods, including biochemical, biological, and biophysical assays of integrin binding, cell adhesion, and protein denaturation. Mutation of the FIII9 synergistic site (PHSRN to PHAAA) in FIII9-10 reduced the binding activity for integrin alpha(5)beta(1) to levels observed for FIII10 alone, but the corresponding mutant in FIII8-9-10 showed no loss of binding activity. Cell adhesion assays also demonstrated enhanced functional activity of constructs containing FIII8. Equilibrium chemical denaturation studies indicated that FIII8 confers conformational stability upon FIII9, but only if the exposed loops, PHSRN and VKNEED on FIII9 and FIII8, respectively, are intact. These results demonstrate that the loss of integrin binding activity, observed upon alteration of the PHSRN synergistic site of FIII9 10, results partly from a loss of conformational stability of FIII9. Our data suggest a mechanism for integrin alpha(5)beta(1)-fibronectin interaction, which in addition to the primary RGD binding event, involves a conformation-sensitive scanning by the integrin for accessible sites on the ligand, whereupon full activation of downstream signaling occurs. PMID- 11500514 TI - Constitutively active mu-opioid receptors inhibit adenylyl cyclase activity in intact cells and activate G-proteins differently than the agonist [D-Ala2,N MePhe4,Gly-ol5]enkephalin. AB - The most convincing evidence demonstrating constitutive activation of mu-opioid receptors is the observation that putative inverse agonists decrease basal G protein activity in membrane preparations. However, it is not clear whether constitutively active receptors in isolated membranes have any physiological relevance in intact cells. GH3 cells expressing mu-opioid receptors (GH3MOR) exhibit higher basal G-protein activity and lower basal cAMP levels than wild type GH3 cells, indicative of constitutively active receptors. This study determined whether alkylation of mu-opioid receptors by the irreversible antagonist beta-funaltrexamine would decrease spontaneous receptor activity in intact cells, revealing constitutive activity. GH3MOR cells were pretreated with increasing concentrations of beta-funaltrexamine followed by functional testing after removal of unbound drug. beta-Funaltrexamine pretreatment produced a concentration-dependent decrease in mu-opioid receptor binding with an IC50 of 0.98 nm and an Emax of 77%. Similar concentrations of beta-funaltrexamine pretreatment produced a half-maximal reduction in basal [35S]GTPgammaS binding, a decrease in basal photolabeling of G-proteins with azidoanilido-[alpha-32P]GTP, and an increase in basal adenylyl cyclase activity in intact cells. Therefore, mu opioid receptors are constitutively active in intact cells, producing stimulation of G-proteins and inhibition of adenylyl cyclase. Importantly, photolabeling of Galpha-subunits with azidoanilido-[alpha-32P]GTP demonstrated that constitutively active mu-opioid receptors activate individual G-proteins differently than the agonist [d-Ala2,N-MePhe4,Gly-ol5]enkephalin. PMID- 11500515 TI - Cloning and characterization of a novel protein kinase that impairs osteoblast differentiation in vitro. AB - The bone morphogenic proteins (BMPs) play a key role in skeletal development and patterning. Using the technique of differential display polymerase chain reaction (ddPCR), we have identified a novel gene whose expression is increased during BMP 2-induced differentiation of the prechondroblastic cell line, MLB13MYC clone 17, to an osteoblastic phenotype. The 6.5-kilobase mRNA recognized by this ddPCR product is increased 10-fold by BMP-2 treatment of the MLB13MYC clone 17 cells. The mRNA recognized by this ddPCR product is also increased as MC3T3-E1 cells recapitulate the program of osteoblast differentiation during prolonged culture. The full-length transcript corresponding to this ddPCR product was cloned from a MLB13MYC clone 17 cell cDNA library. Analysis of the deduced amino acid sequence demonstrated that this gene encodes a novel 126-kDa putative serine/threonine protein kinase containing a nuclear localization signal. The kinase domain, expressed in Escherichia coli, is capable of autophosphorylation as well as phosphorylation of myelin basic protein. The gene was, therefore, named BIKe (BMP 2-Inducible Kinase). The BIKe nuclear localization signal is able to direct green fluorescent protein to the nucleus in transfected COS-7 cells. When stably expressed in MC3T3-E1 cells, BIKe significantly decreases alkaline phosphatase activity and osteocalcin mRNA levels and retards mineral deposition relative to vector control. This novel kinase, therefore, is likely to play an important regulatory role in attenuating the program of osteoblast differentiation. PMID- 11500516 TI - Multiple ErbB-2/Neu Phosphorylation Sites Mediate Transformation through Distinct Effector Proteins. AB - Amplification of the type I receptor tyrosine kinase ErbB-2 (HER2/Neu) is observed in 20-30% of human mammary carcinomas, correlating with a poor clinical prognosis. We have previously demonstrated that four (Tyr(1144), Tyr(1201), Tyr(1226/1227), or Tyr(1253)) of the five known Neu/ErbB-2 autophosphorylation sites can independently mediate transforming signals. The transforming potential of at least two of these autophosphorylation sites (Tyr(1144) and Tyr(1226/1227)) has been further correlated with their ability to associate with Grb2 and Shc adapter proteins, respectively. To confirm the specificity of these interactions, we have created a series of second site mutants in these phosphorylation sites. The results showed that Grb2 recruitment to site 1144 is absolutely required for transforming signal from this autophosphorylation site, whereas association of Shc-mediated transformation is dependent on conservation of the NPXY motif spanning Tyr(1227). A stretch of amino acid identity around tyrosines 1201 (ENPEYLTP)and 1253 (ENPEYLDL) exists, and mutation of key residues within this motif reveals distinct requirements for an intact protein tyrosine-binding protein (NPXY). We show that DOK-R, a protein tyrosine-binding site-containing protein implicated in Ras signaling, interacts with Neu/ErbB-2 at Tyr(1253) as do two unidentified proteins, p150 and p34, the latter correlating with transformation. Together these data argue that ErbB-2/Neu is capable of mediating transformation through distinct effector pathways. PMID- 11500517 TI - Peroxisomal straight-chain Acyl-CoA oxidase and D-bifunctional protein are essential for the retroconversion step in docosahexaenoic acid synthesis. AB - Docosahexaenoic acid (DHA, C22:6n-3) is essential for normal brain and retinal development. The nature and subcellular location of the terminal steps in DHA biosynthesis have been controversial. Rather than direct Delta4-desaturation of C22:5n-3, it has been proposed that this intermediate is elongated to C24:5n-3, desaturated to C24:6n-3, and "retroconverted" to DHA via peroxisomal beta oxidation. However, this hypothesis has recently been challenged. The goal of this study was to determine the mechanism and specific enzymes required for the retroconversion step in human skin fibroblasts. Cells from patients with deficiencies of either acyl-CoA oxidase or D-bifunctional protein, the first two enzymes of the peroxisomal straight-chain fatty acid beta-oxidation pathway, exhibited impaired (5-20% of control) conversion of either [1-14C]18:3n-3 or [1 14C]22:5n-3 to DHA as did cells from peroxisome biogenesis disorder patients comprising eight distinct genotypes. In contrast, normal DHA synthesis was observed in cells from patients with rhizomelic chondrodysplasia punctata, Refsum disease, X-linked adrenoleukodystrophy, and deficiency of mitochondrial medium- or very long-chain acyl-CoA dehydrogenase. Acyl-CoA oxidase-deficient cells accumulated 2-5 times more radiolabeled C24:6n-3 than did controls. Our data are consistent with the retroconversion hypothesis and demonstrate that peroxisomal beta-oxidation enzymes acyl-CoA oxidase and D-bifunctional protein are essential for this process in human skin fibroblasts. PMID- 11500518 TI - Oleoyl-CoA is the major de novo product of stearoyl-CoA desaturase 1 gene isoform and substrate for the biosynthesis of the Harderian gland 1-alkyl-2,3 diacylglycerol. AB - 1-Alkyl-2,3-diacylglycerol (ADG) is a unique neutral lipid found in the eyeball associated Harderian gland (HG) of the mouse and acts as a lubricant to facilitate eyelid movement. We found that the HG of the mice with a disruption in the gene for stearoyl-CoA desaturase 1 (SCD1) (SCD1-/-) is deficient in ADG. The amount of C20:1n-9, which is a major fatty acid of ADG, was reduced by greater than 90% despite normal elongase enzyme activity proposed to elongate it from C18:1n-9. HG from SCD1-/- mice exhibited high desaturase activity toward C16:0 CoA as substrate but had very low desaturase activity toward C18:0-CoA. Feeding diets containing high levels of oleate to the SCD1-/- mice did not increase the levels of C18:1n-9 or C20:1n-9 in the HG and failed to restore the ADG to the levels found in the HG of the wild-type mouse. De novo ADG synthesis as measured by the incorporation of [(3)H]glycerol and [(14)C]glucose was high in the SCD1+/+ mouse but was reduced by greater than 90% in the HG of SCD1-/- mouse. The deficiencies in the levels of ADG and C20:1n-9 were not compensated for by the expression of SCD2 and SCD3 isoforms in the HG of the SCD1-/- mouse. These observations demonstrate that SCD1-synthesized oleoyl-CoA is a major substrate required for the biosynthesis of normal levels of ADG and that the SCD isoforms present in the HG have different substrate specificity. PMID- 11500519 TI - RyR3 amplifies RyR1-mediated Ca(2+)-induced Ca(2+) release in neonatal mammalian skeletal muscle. AB - The neonatal mammalian skeletal muscle contains both type 1 and type 3 ryanodine receptors (RyR1 and RyR3) located in the sarcoplasmic reticulum membrane. An allosteric interaction between RyR1 and dihydropyridine receptors located in the plasma membrane mediates voltage-induced Ca(2+) release (VICR) from the sarcoplasmic reticulum. RyR3, which disappears in adult muscle, is not involved in VICR, and the role of the transiently expressed RyR3 remains elusive. Here we demonstrate that RyR1 participates in both VICR and Ca(2+)-induced Ca(2+) release (CICR) and that RyR3 amplifies RyR1-mediated CICR in neonatal skeletal muscle. Confocal measurements of intracellular Ca(2+) in primary cultured mouse skeletal myotubes reveal active sites of Ca(2+) release caused by peripheral coupling between dihydropyridine receptors and RyR1. In myotubes lacking RyR3, the peripheral VICR component is unaffected, and RyR1s alone are able to support inward CICR propagation in most cells at an average speed of approximately 190 microm/s. With the co-presence of RyR1 and RyR3 in wild-type cells, unmitigated radial CICR propagates at 2,440 microm/s. Because neonatal skeletal muscle lacks a well developed transverse tubule system, the RyR3 reinforcement of CICR seems to ensure a robust, uniform, and synchronous activation of Ca(2+) release throughout the cell body. Such functional interplay between RyR1 and RyR3 can serve important roles in Ca(2+) signaling of cell differentiation and muscle contraction. PMID- 11500520 TI - Decreased cardiolipin synthesis corresponds with cytochrome c release in palmitate-induced cardiomyocyte apoptosis. AB - Apoptosis has been identified recently as a component of many cardiac pathologies. However, the potential triggers of programmed cell death in the heart and the involvement of specific metabolic pathway(s) are less well characterized. Detachment of cytochrome c from the mitochondrial inner membrane is a necessary first step for cytochrome c release into the cytosol and initiation of apoptosis. The saturated long chain fatty acid, palmitate, induces apoptosis in rat neonatal cardiomyocytes and diminishes content of the mitochondrial anionic phospholipid, cardiolipin. These changes are accompanied by 1) acyl chain saturation of phosphatidic acid and phosphatidylglycerol, 2) large increases in the levels of these two phospholipids, and 3) a decline in cardiolipin synthesis. Although cardiolipin synthase activity is unchanged, saturated phosphatidylglycerol is a poor substrate for this enzyme. Under these conditions, decreased cardiolipin synthesis and release of cytochrome c are directly and significantly correlated. The results suggest that phosphatidylglycerol saturation and subsequent decreases in cardiolipin affect the association of cytochrome c with the inner mitochondrial membrane, directly influencing the pathway to cytochrome c release and subsequent apoptosis. PMID- 11500521 TI - The extracellular calcium-sensing receptor is required for calcium-induced differentiation in human keratinocytes. AB - In cultured keratinocytes, the acute increase of the extracellular calcium concentration above 0.03 mM leads to a rapid increase in intracellular calcium concentration ([Ca(2+)]i) and inositol trisphosphate production and, subsequently, to the expression of differentiation-related genes. Previous studies demonstrated that human keratinocytes express the full-length extracellular calcium-sensing receptor (CaR) and an alternatively spliced variant lacking exon 5 and suggested their involvement in calcium regulation of keratinocyte differentiation. To understand the role of the CaR, we transfected keratinocytes with an antisense human CaR cDNA construct and examined its impact on calcium signaling and calcium-induced differentiation. The antisense CaR cDNA significantly reduced the protein level of endogenous CaRs. These cells displayed a marked reduction in the rise in [Ca(2+)]i in response to extracellular calcium or to NPS R-467, a CaR activator, whereas the ATP-evoked rise in [Ca(2+)]i was not affected. Calcium-induced inhibition of cell proliferation and calcium stimulated expression of the differentiation markers involucrin and transglutaminase were also blocked by the antisense CaR cDNA. When cotransfected with luciferase reporter vectors containing either the involucrin or transglutaminase promoter, the antisense CaR cDNA suppressed the calcium stimulated promoter activities. These results indicate that CaR is required for mediating calcium signaling and calcium-induced differentiation in keratinocytes. PMID- 11500522 TI - Influence of total vs. visceral fat on insulin action and secretion in African American and white children. AB - OBJECTIVE: To examine whether total body fat (FAT) in general or visceral fat (VFAT) in particular is associated with greater metabolic risk in white and African American children. RESEARCH METHODS AND PROCEDURES: A total of 68 white and 51 African American children had measures of insulin sensitivity (Si) and acute insulin response (AIR) by a frequently sampled intravenous glucose tolerance test, total body fat by DXA and abdominal fat distribution (visceral vs. subcutaneous) by computed tomography. The influence of FAT and VFAT on insulin parameters were examined by comparing subgroups of children with high or low FAT vs. high or low VFAT and by multiple regression analysis. RESULTS: In whites, fasting insulin, Si, and AIR were significantly influenced by FAT, but not VFAT (e.g., for Si, 9.8 +/- 0.8 in low FAT vs. 4.6 +/- 0.7 x 10( 4)/min/[microIU/mL[ in high FAT, p < 0.05; 6.8 +/- 0.7 in low VFAT vs. 7.5 +/- 0.8 x 10(-4)/min/[microIU/mL] in high VFAT, p > 0.1). In African Americans, fasting insulin and Si were also primarily influenced by FAT (e.g., for Si, 4.9 +/- 0.4 in low FAT vs. 2.8 +/- 0.5 x 10(-4)/min/[microIU/mL] in high FAT, p < 0.05) but not by VFAT, and there were no significant effects of either fat compartment on AIR. In multiple regression analysis, Si was significantly influenced by FAT (negative effect), ethnicity (lower in African Americans), and gender (lower in females), whereas fasting insulin was significantly influenced by VFAT (positive effect), ethnicity (higher in African Americans), and fat free mass (positive effect). DISCUSSION: Body fat in general is the predominant factor influencing Si, but VFAT may have additional effects on fasting insulin. The lack of major effects of VFAT on Si in children may be explained by lower levels of VFAT or because VFAT affects aspects of whole body insulin action that are not measured by the minimal model. PMID- 11500523 TI - Total and regional fat and serum cardiovascular disease risk factors in lean and obese children and adolescents. AB - OBJECTIVE: This study was conducted to evaluate the association of total and central adiposity with serum cardiovascular disease (CVD) risk factors in lean and obese Portuguese children and adolescents. RESEARCH METHODS AND PROCEDURES: A total of 87 girls (13.2 +/- 1.6 years old, 29.9 +/- 6.4% body fat [mean +/- SD]) and 72 boys (13.2 +/- 1.6 years old, 20.8 +/- 9.9% body fat) volunteered for the study. Whole-body composition and fat distribution, from DXA and anthropometry, and serum lipids, lipoproteins, and apolipoproteins were evaluated. RESULTS: The sum of three trunk skinfolds (STS) was highly correlated with total trunk fat mass measured by DXA (p < 0.001). Body mass index, DXA-measured percentage of body fat, trunk fat mass, STS, and the waist-to-height ratio were generally found to be associated with triacylglycerol, the ratio of total cholesterol (TC) to high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), and apolipoprotein B levels, (significant age-adjusted r between 0.16 and 0.27, p < 0.05). Body mass index, STS, and the waist circumference were also associated with HDL-C (p < 0.05), whereas no body composition variable significantly correlated with TC or apolipoproteins A-I. The STS was significantly correlated with HDL-C (p < 0.01), TC/HDL-C (p < 0.05), and apolipoproteins A-I (p < 0.05) independently of whole-body fatness. Obese subjects (n = 73) had higher TC, LDL-C, TC/HDL-C, and apolipoprotein B than did non-obese subjects (n = 86), and significant associations between central adiposity and some lipid variables (triacylglycerol and HDL-C) were found in obese children and adolescents that were not present in leaner individuals. DISCUSSION: DXA- and anthropometry-based whole-body and central fat measures are associated with serum CVD risk factors in Portuguese boys and girls. Obese children and adolescents have a poorer lipid profile than do their leaner counterparts. Trunk skinfolds, which are easy to obtain even in large samples, predict CVD risk factors to the same extent as DXA-based variables, in some cases, independently of total fatness. PMID- 11500524 TI - Total and central obesity among elderly Hispanics and the association with Type 2 diabetes. AB - OBJECTIVE: To report the prevalence of total and central obesity in a representative sample of Puerto Rican and Dominican elders in Massachusetts, to compare them with a neighborhood-based group of non-Hispanic white elders, and to examine associations of obesity indices with the presence of type 2 diabetes. RESEARCH METHODS AND PROCEDURES: We examined the prevalence of overweight, obesity, and central obesity in 596 Hispanics of Caribbean origin, ages 60 to 92 years, and 239 non-Hispanic whites, and tested linear and logistic regression models to determine associations among body mass index (BMI), waist circumference (WC), and diabetes. RESULTS: Obesity (BMI > or = 30 kg/m(2)) was prevalent among all ethnic groups, ranging from 17% to 29% for Dominican and Puerto Rican men, respectively, and from 29% to 40% for non-Hispanic white and Dominican women, respectively. These differences were not statistically significant. Among Hispanic men and women, diabetes was prevalent across all BMI and WC categories but tended to be greatest among those with BMI of 25 to 29 kg/m(2) (41% to 43%). In contrast, diabetes was most prevalent in the obese group (36% to 45%) of non Hispanic whites. Both BMI and WC were associated with the presence of diabetes, but the coefficients were greater for non-Hispanic whites than for Hispanics. DISCUSSION: Caribbean Hispanics and non-Hispanic whites living in the same Massachusetts localities had high prevalences of overweight and obesity. Total and central obesity exerted a differential effect on the presence of diabetes among ethnic groups; for Hispanics, diabetes was prevalent even among non-obese individuals, whereas for non-Hispanic white women, the prevalence of diabetes was strongly associated with total and central obesity. Additional research is needed to investigate the factors associated with the differential effect of obesity on the prevalence of type 2 diabetes among Hispanic and non-Hispanic white elders. PMID- 11500525 TI - Leptin levels are associated with fat oxidation and dietary-induced weight loss in obesity. AB - OBJECTIVE: To examine the relationship between fasting plasma leptin and 24-hour energy expenditure (EE), substrate oxidation, and spontaneous physical activity (SPA) in obese subjects before and after a major weight reduction compared with normal weight controls. To test fasting plasma leptin, substrate oxidations, and SPA as predictive markers of success during a standardized weight loss intervention. RESEARCH METHODS AND PROCEDURES: Twenty-one nondiabetic obese (body mass index: 33.9 to 43.8 kg/m(2)) and 13 lean (body mass index: 20.4 to 24.7 kg/m(2)) men matched for age and height were included in the study. All obese subjects were reexamined after a mean weight loss of 19.2 kg (95% confidence interval: 15.1-23.4 kg) achieved by 16 weeks of dietary intervention followed by 8 weeks of weight stability. Twenty-four-hour EE and substrate oxidations were measured by whole-body indirect calorimetry. SPA was assessed by microwave radar. RESULTS: In lean subjects, leptin adjusted for fat mass (FM) was correlated to 24 hour EE before (r = -0.56, p < 0.05) but not after adjustment for fat free mass. In obese subjects, leptin correlated inversely with 24-hour and resting nonprotein respiratory quotient (r = -0.47, p < 0.05 and r = -0.50, p < 0.05) both before and after adjustments for energy balance. Baseline plasma leptin concentration, adjusted for differences in FM, was inversely related to the size of weight loss after 8 weeks (r = -0.41, p = 0.07), 16 weeks (r = -0.51, p < 0.05), and 24 weeks (r = -0.50, p < 0.05). DISCUSSION: The present study suggests that leptin may have a stimulating effect on fat oxidation in obese subjects. A low leptin level for a given FM was associated with a greater weight loss, suggesting that obese subjects with greater leptin sensitivities are more successful in reducing weight. PMID- 11500526 TI - The effects of leptin administration in non-obese human subjects. AB - OBJECTIVE: Body fatness is partly under hypothalamic control with effector limbs that include the endocrine system and the autonomic nervous system (ANS). In previous studies of both obese and never-obese subjects, we have shown that weight increase leads to increased sympathetic and decreased parasympathetic activity, whereas weight decrease leads to decreased sympathetic and increased parasympathetic activity. We now report on the effect of leptin, independent of weight change, on the ANS. RESEARCH METHODS AND PROCEDURES: Normal weight males (ages 20-40 years) were fed a solid food diet, measured carefully to maintain body weight, for 3 weeks, as inpatients at the Rockefeller University General Clinical Research Center. In a single-blind, 22-day, placebo/drug/placebo design, six subjects received leptin 0.3 mg/kilogram subcutaneously for 6 days. ANS measures of amount of parasympathetic control and sympathetic control of heart period (interbeat interval) were made by sequential pharmacological blockade with intravenous atropine and esmolol. Norepinephrine, dopamine, and epinephrine levels in 24-hour urine collections were also measured as well as resting metabolic rate. RESULTS: Sufficient food intake maintained constant body weight in all subjects. There was no evidence that leptin administration led to changes in energy metabolism sufficient to require additional food intake or to alter resting metabolic rate. Likewise, leptin administration did not alter autonomic activity. Parasympathetic control and sympathetic control, as well as the urinary catecholamines, were not significantly affected by leptin administration. Glucose and insulin levels were increased by food intake as expected, but leptin had no affect on these levels before or after food intake. DISCUSSION: ANS responses to changes in energy metabolism found when food intake and body weight are altered were not found in these never-obese subjects given leptin for 6 days. Although exogenous leptin administration has profound effects on food intake and energy metabolism in animals genetically deprived of leptin, we found it to have no demonstrable effect on energy metabolism in never-obese humans. The effects of longer periods of administration to obese individuals and to those who have lost weight demand additional investigation. PMID- 11500527 TI - Neck circumference as a simple screening measure for identifying overweight and obese patients. AB - OBJECTIVE: There are numerous methods of assessing overweight and obesity. We undertook an observational study to test a method of identifying overweight or obese patients solely by measuring the circumference of the neck. RESEARCH METHODS AND PROCEDURES: A test sample and a second validation sample included 979 subjects (460 men and 519 women), who visited a family medicine clinic in a southern Israeli urban district for any reason between the randomly chosen months of January and September 1998. Main outcome included neck, waist, and hip circumferences; body mass index (BMI); and waist:hip ratio measures. RESULTS: Pearson's correlation coefficients indicated a significant association between neck circumference (NC) and: BMI (men, r = 0.83; women, r = 0.71; each, p < 0.0001), age (men, r = 0.33; women, r = 0.36; each, p < 0.0001), weight (men, r = 0.7; women, r = 0.81; each, p < 0.0001), waist circumference (men, r = 0.86; women, r = 0.85; each, p < 0.0001), hip circumference (men, r = 0.62; women, r = 0.56; each, p < 0.0001), and waist:hip ratio (men, r = 0.66; women, r = 0.87; each, p < 0.0001). NC > or=37 cm for men and > or =34 cm for women were the best cutoff levels for determining the subjects with BMI > or =25.0 kg/m(2) using the receiver output curve analysis. In the validation unrelated group, the test characteristics were excellent with 98% sensitivity, 89% specificity, and 94% accuracy for men, and 100% sensitivity, 98% specificity, and 99% accuracy for women. NC > or =39.5 cm for men and > or =36.5 cm for women were the best cutoff levels for determining the subjects with BMI > or =30 kg/m(2) using the receiver output curve analysis. In the validation unrelated group, the test characteristics were excellent with 93% sensitivity, 90% specificity, and 91% accuracy for men, and 93% sensitivity, 98% specificity, and 97% accuracy for women. DISCUSSION: NC measurement is a simple and time-saving screening measure that can be used to identify overweight and obese patients. Men with NC <37 cm and women with NC <34 cm are not to be considered overweight. Patients with NC > or =37 cm for men and > or =34 cm for women require additional evaluation of overweight or obesity status. PMID- 11500528 TI - Ability of young women to recall past body size and age at menarche. AB - OBJECTIVE: The ability of young women (n = 132, average age 17 years) to recall body size and age at menarche was examined. The use of body silhouettes to assist women in recalling their body size at menarche and to represent their current body size was also evaluated. RESEARCH METHODS AND PROCEDURES: Subjects, who previously participated in a cohort study, were asked to recall height and weight at the time of menarche, to select body silhouettes that best reflected their body shape at the time of menarche and their current body shape, and to recall age at menarche. Two sets of body silhouettes were developed, one representative of an adult body shape and another representative of an adolescent body shape. RESULTS: Pearson correlation coefficients between the adult and adolescent body figures and actual body mass index (BMI; kg/m(2)) at the time of menarche were not significantly different (r = 0.82 for adult figures vs. r = 0.72 for adolescent figures, p > 0.05). The correlation between actual BMI at the time of menarche and body silhouette (r = 0.77, all subjects) was similar to the correlation between actual and recalled BMI at the time of menarche (r = 0.83) as well as the correlation between current BMI and current body silhouette (r = 0.75). Recalled and actual ages at menarche were highly correlated (r = 0.83). DISCUSSION: The recall of body shape was considered to be a less precise measure of body size than asking about height and weight, but use of body silhouettes may offer advantages in certain situations. PMID- 11500529 TI - Metabolic and hemodynamic response of adipose tissue to angiotensin II. AB - OBJECTIVE: Recent studies have revealed the presence of a local renin-angiotensin system in adipose tissue. To examine the possible role of this system in adipose tissue, we performed microdialysis studies on the effect of angiotensin II (Ang II) on blood flow and metabolism in abdominal subcutaneous adipose tissue (aSAT) and femoral subcutaneous adipose tissue (fSAT) in young healthy men. RESEARCH METHODS AND PROCEDURES: Using the microdialysis technique, two different protocols were run perfusion with Ringer's solution + 50 mM ethanol with the subsequent addition of 125, 250, and 500 microg/liter Ang II (n = 8) and Ringers's solution + 50 mM ethanol with the subsequent addition of isoproterenol (1 microM) alone and in combination with 500 microg/liter Ang II (n = 6). Dialysate concentrations of ethanol, glycerol, glucose, and lactate were measured for estimating blood flow (ethanol dilution technique), lipolysis, and glycolysis, respectively. RESULTS: Perfusion with Ang II resulted in a dose dependent decrease in blood flow (fSAT > aSAT), lipolysis (fSAT > aSAT), and glucose uptake (fSAT = aSAT). Isoproterenol increased blood flow and lipolysis at both sites and those effects could be returned to baseline values by the addition of Ang II in aSAT but not fSAT. DISCUSSION: In conclusion, our data indicate that in addition to its well-known vasoconstricting effect, Ang II inhibits lipolysis in adipose tissue, whereby femoral fat depots seem to be more sensitive to this effect than abdominal depots. PMID- 11500530 TI - Hyperphagia as a mediator of renal disease initiation in obese Zucker rats. AB - OBJECTIVE: We sought to determine whether prevention of overeating would block the very earliest manifestations of renal injury in young obese Zucker rats (OZRs). RESEARCH METHODS AND PROCEDURES: Three groups of rats were studied, obese (fa/fa) Zucker rats and lean (Fa/Fa). Zucker controls were allowed to feed ad libitum, whereas a group of obese (fa/fa) Zucker rats was pair-fed to the lean group. Urine albumin and serum lipids were studied weekly from 6 to 10 weeks of age. Renal pathology and renal glomerular gene expression were examined when the rats were killed at 10 weeks of age. RESULTS: Obese rats fed ad libitum developed significant albuminuria by 6 weeks of age, increasing at each subsequent time point. This increase was completely blocked by pair-feeding. Serum triglycerides were significantly increased in obese rats fed ad libitum vs. the other groups. Urine albumin correlated significantly with both body weight and serum triglyceride level. Renal histopathology was normal in all groups. Analysis of gene expression of glomerular proteins by reverse transcriptase-polymerase chain reaction revealed that pair-feeding attenuated the increased expression of glomerular desmin, fibronectin, and the 92-kDa collagenase that was seen in obese animals fed ad libitum. DISCUSSION: Prevention of overeating in young OZR normalizes albuminuria and attenuates the pathogenic alterations in glomerular gene expression seen at the initiation of renal disease in obese animals allowed to feed ad libitum. This model may be relevant for studying the early end-organ effects of obesity. PMID- 11500531 TI - Changes of adiposity in response to vitamin A status correlate with changes of PPAR gamma 2 expression. AB - OBJECTIVE: To gain insight into the in vivo modulation of the expression of the adipogenic transcription factors PPAR gamma 2, C/EBP alpha, and ADD1/SREBP1c by retinoids and its relationship with whole-body adiposity. RESEARCH METHODS AND PROCEDURES: Three-week-old mice were fed with standard chow or a vitamin A deficient diet for 10 weeks. During the 4 days immediately before they were killed, the animals were treated either with all-trans retinoic acid (tRA; 100 mg/kg per day, subcutaneously) or vehicle. The specific levels of the mRNAs for the three transcription factors were analyzed in epididymal white adipose tissue (eWAT) and inguinal white adipose tissue and in brown adipose tissue (BAT). Other parameters determined were leptin and UCP2 levels in white adipose tissue depots, total cholesterol and triglyceride serum levels, energy intake, body weight, and adiposity. RESULTS: Vitamin A-deficient diet feeding led to a marked increase of adiposity and to a small increase of body weight. Hypertrophy of white adipose tissue depots correlated with enhanced PPAR gamma 2 expression. Hypertrophy of BAT, in contrast, correlated with a decrease of PPAR gamma 2 expression that may contribute to the known reduced thermogenic potential of BAT under conditions of vitamin A restriction. Treatment with tRA triggered a reduction of adiposity and body weight that correlated with a down-regulation of PPAR gamma 2 expression in all adipose tissues. The effects of tRA were more pronounced in eWAT, where C/EBP alpha and ADD1/SREBP1c levels were also reduced. The response to tRA was impaired in the eWAT and BAT of animals fed the vitamin A-deficient diet. DISCUSSION: The results emphasize the importance of retinoids as physiological regulators of adipose tissue development and function in intact animals. PMID- 11500533 TI - Samples from fifty years of career decisions. PMID- 11500534 TI - A life in science, editing, and writing. PMID- 11500535 TI - Lignin formation in plants. The dilemma of linkage specificity. PMID- 11500536 TI - The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants. AB - Major intrinsic proteins (MIPs) facilitate the passive transport of small polar molecules across membranes. MIPs constitute a very old family of proteins and different forms have been found in all kinds of living organisms, including bacteria, fungi, animals, and plants. In the genomic sequence of Arabidopsis, we have identified 35 different MIP-encoding genes. Based on sequence similarity, these 35 proteins are divided into four different subfamilies: plasma membrane intrinsic proteins, tonoplast intrinsic proteins, NOD26-like intrinsic proteins also called NOD26-like MIPs, and the recently discovered small basic intrinsic proteins. In Arabidopsis, there are 13 plasma membrane intrinsic proteins, 10 tonoplast intrinsic proteins, nine NOD26-like intrinsic proteins, and three small basic intrinsic proteins. The gene structure in general is conserved within each subfamily, although there is a tendency to lose introns. Based on phylogenetic comparisons of maize (Zea mays) and Arabidopsis MIPs (AtMIPs), it is argued that the general intron patterns in the subfamilies were formed before the split of monocotyledons and dicotyledons. Although the gene structure is unique for each subfamily, there is a common pattern in how transmembrane helices are encoded on the exons in three of the subfamilies. The nomenclature for plant MIPs varies widely between different species but also between subfamilies in the same species. Based on the phylogeny of all AtMIPs, a new and more consistent nomenclature is proposed. The complete set of AtMIPs, together with the new nomenclature, will facilitate the isolation, classification, and labeling of plant MIPs from other species. PMID- 11500537 TI - Overexpression of KNAT1 in lettuce shifts leaf determinate growth to a shoot-like indeterminate growth associated with an accumulation of isopentenyl-type cytokinins. AB - Leaves are specialized organs characterized by defined developmental destiny and determinate growth. The overexpression of Knotted1-like homeobox genes in different species has been shown to alter leaf shape and development, but a definite role for this class of genes remains to be established. Transgenics that overexpress Knotted1-like genes present some traits that are characteristic of altered cytokinin physiology. Here we show that lettuce (Lactuca sativa) leaves that overexpress KNAT1, an Arabidopsis kn1-like gene, acquire characteristics of indeterminate growth typical of the shoot and that this cell fate change is associated with the accumulation of specific types of cytokinins. The possibility that the phenotypic effects of KNAT1 overexpression may arise primarily from the modulation of local ratios of different cytokinins is discussed. PMID- 11500538 TI - Aluminum inhibits the H(+)-ATPase activity by permanently altering the plasma membrane surface potentials in squash roots. AB - Although aluminum (AL) toxicity has been widely studied in monocotyledonous crop plants, the mechanism of Al impact on economically important dicotyledonous plants is poorly understood. Here, we report the spatial pattern of Al-induced root growth inhibition, which is closely associated with inhibition of H(+) ATPase activity coupled with decreased surface negativity of plasma membrane (PM) vesicles isolated from apical 5-mm root segments of squash (Cucurbita pepo L. cv Tetsukabuto) plants. High-sensitivity growth measurements indicated that the central elongation zone, located 2 to 4 mm from the tip, was preferentially inhibited where high Al accumulation was found. The highest positive shifts (depolarization) in zeta potential of the isolated PM vesicles from 0- to 5-mm regions of Al-treated roots were corresponded to pronounced inhibition of H(+) ATPase activity. The depolarization of PM vesicles isolated from Al-treated roots in response to added Al in vitro was less than that of control roots, suggesting, particularly in the first 5-mm root apex, a tight Al binding to PM target sites or irreversible alteration of PM properties upon Al treatment to intact plants. In line with these data, immunolocalization of H(+)-ATPase revealed decreases in tissue-specific H(+)-ATPase in the epidermal and cortex cells (2--3 mm from tip) following Al treatments. Our report provides the first circumstantial evidence for a zone-specific depolarization of PM surface potential coupled with inhibition of H(+)-ATPase activity. These effects may indicate a direct Al interaction with H(+)-ATPase from the cytoplasmic side of the PM. PMID- 11500539 TI - Molybdenum sequestration in Brassica species. A role for anthocyanins? AB - To elucidate plant mechanisms involved in molybdenum (Mo) sequestration and tolerance, Brassica spp. seedlings were supplied with molybdate, and the effects on plant physiology, morphology, and biochemistry were analyzed. When supplied with (colorless) molybdate Indian mustard (Brassica juncea) seedlings accumulated water-soluble blue crystals in their peripheral cell layers. Energy dispersive x ray analysis showed that Mo accumulated predominantly in the vacuoles of the epidermal cells. Therefore, the blue crystals are likely to be a Mo compound. The x-ray absorption spectrum of the plant-accumulated Mo was different than that for molybdate, indicating complexation with a plant molecule. Because the blue compound was water soluble and showed a pH-dependent color change, possible involvement of anthocyanins was investigated. An anthocyanin-less mutant of Brassica rapa ("fast plants") was compared with varieties containing normal or high anthocyanin levels. The anthocyanin-less mutant did not show accumulation of a blue compound when supplied with molybdate. In the anthocyanin-containing varieties, the blue compound colocalized with anthocyanins in the peripheral cell layers. Mo accumulation by the three B. rapa varieties was positively correlated with anthocyanin content. Addition of molybdate to purified B. rapa anthocyanin resulted in an in vitro color change from pink to blue. Therefore, Mo appears to be sequestered in vacuoles of the peripheral cell layers of Brassica spp. as a blue compound, probably a Mo-anthocyanin complex. PMID- 11500540 TI - A new C-type cyclin-dependent kinase from tomato expressed in dividing tissues does not interact with mitotic and G1 cyclins. AB - Cyclin-dependent kinases (CDKs) form a conserved superfamily of eukaryotic serine threonine protein kinases whose activity requires the binding of a cyclin protein. CDKs are involved in many aspects of cell biology and notably in the regulation of the cell cycle. Three cDNAs encoding a C-type CDK, and a member of each B-type CDK subfamily, were isolated from tomato (Lycopsersicon esculentum Mill.) and designated Lyces;CDKC;1 (accession no. AJ294903), Lyces; CDKB1;1 (accession no. AJ297916), and Lyces;CDKB2;1 (accession no. AJ297917). The predicted amino acid sequences displayed the characteristic PITAIRE (CDKC), PPTALRE (CDKB1), and PPTTLRE (CDKB2) motives in the cyclin-binding domain, clearly identifying the type of CDK. The accumulation of all transcripts was associated preferentially with dividing tissues in developing tomato fruit and vegetative organs. In contrast to that of CDKA and CDKBs, the transcription pattern of Lyces;CDKC;1 was shown to be independent of hormone and sugar supply in tomato cell suspension cultures and excised roots. This observation, together with the absence of a patchy expression profile in in situ hybridization experiments, suggests a non-cell cycle regulation of Lyces;CDKC;1. Using a two hybrid assay, we showed that Lyces;CDKC;1 did not interact with mitotic and G1 cyclins. The role of plant CDKCs in the regulation of cell division and differentiation is discussed with regard to the known function of their animal counterparts. PMID- 11500541 TI - Efficient prenylation by a plant geranylgeranyltransferase-I requires a functional CaaL box motif and a proximal polybasic domain. AB - Geranylgeranyltransferase-I (GGT-I) is a heterodimeric enzyme that shares a common alpha-subunit with farnesyltransferase (FTase) and has a distinct beta subunit. GGT-I preferentially modifies proteins, which terminate in a CaaL box sequence motif. Cloning of Arabidopsis GGT-I beta-subunit (AtGGT-IB) was achieved by a yeast (Saccharomyces cerevisiae) two-hybrid screen, using the tomato (Lycopersicon esculentum) FTase alpha-subunit (FTA) as bait. Sequence and structure analysis revealed that the core active site of GGT-I and FTase are very similar. AtGGT-IA/FTA and AtGGT-IB were co-expressed in baculovirus-infected insect cells to obtain recombinant protein that was used for biochemical and molecular analysis. The recombinant AtGGT-I prenylated efficiently CaaL box fusion proteins in which the a(2) position was occupied by an aliphatic residue, whereas charged or polar residues at the same position greatly reduced the efficiency of prenylation. A polybasic domain proximal to the CaaL box motif induced a 5-fold increase in the maximal reaction rate, and increased the affinity of the enzyme to the protein substrate by an order of magnitude. GGT-I retained high activity in a temperature range between 24 degrees C and 42 degrees C, and showed increased activity rate at relatively basic pH values of 7.9 and 8.5. Reverse transcriptase-polymerase chain reaction, protein immuno-blots, and transient expression assays of green fluorescent protein fusion proteins show that GGT-IB is ubiquitously expressed in a number of tissues, and that expression levels and protein activity were not changed in mutant plants lacking FTase beta subunit. PMID- 11500542 TI - Numeric simulation of plant signaling networks. AB - Plants have evolved an intricate signaling apparatus that integrates relevant information and allows an optimal response to environmental conditions. For instance, the coordination of defense responses against pathogens involves sophisticated molecular detection and communication systems. Multiple protection strategies may be deployed differentially by the plant according to the nature of the invading organism. These responses are also influenced by the environment, metabolism, and developmental stage of the plant. Though the cellular signaling processes traditionally have been described as linear sequences of events, it is now evident that they may be represented more accurately as network-like structures. The emerging paradigm can be represented readily with the use of Boolean language. This digital (numeric) formalism allows an accurate qualitative description of the signal transduction processes, and a dynamic representation through computer simulation. Moreover, it provides the required power to process the increasing amount of information emerging from the fields of genomics and proteomics, and from the use of new technologies such as microarray analysis. In this review, we have used the Boolean language to represent and analyze part of the signaling network of disease resistance in Arabidopsis. PMID- 11500543 TI - Hydrogen peroxide is involved in abscisic acid-induced stomatal closure in Vicia faba. AB - One of the most important functions of the plant hormone abscisic acid (ABA) is to induce stomatal closure by reducing the turgor of guard cells under water deficit. Under environmental stresses, hydrogen peroxide (H(2)O(2)), an active oxygen species, is widely generated in many biological systems. Here, using an epidermal strip bioassay and laser-scanning confocal microscopy, we provide evidence that H(2)O(2) may function as an intermediate in ABA signaling in Vicia faba guard cells. H(2)O(2) inhibited induced closure of stomata, and this effect was reversed by ascorbic acid at concentrations lower than 10(-5) M. Further, ABA induced stomatal closure also was abolished partly by addition of exogenous catalase (CAT) and diphenylene iodonium (DPI), which are an H(2)O(2) scavenger and an NADPH oxidase inhibitor, respectively. Time course experiments of single cell assays based on the fluorescent probe dichlorofluorescein showed that the generation of H(2)O(2) was dependent on ABA concentration and an increase in the fluorescence intensity of the chloroplast occurred significantly earlier than within the other regions of guard cells. The ABA-induced change in fluorescence intensity in guard cells was abolished by the application of CAT and DPI. In addition, ABA microinjected into guard cells markedly induced H(2)O(2) production, which preceded stomatal closure. These effects were abolished by CAT or DPI micro-injection. Our results suggest that guard cells treated with ABA may close the stomata via a pathway with H(2)O(2) production involved, and H(2)O(2) may be an intermediate in ABA signaling. PMID- 11500544 TI - Phospholipase D and phosphatidic acid-mediated generation of superoxide in Arabidopsis. AB - Phospholipase D (PLD), which hydrolyzes phospholipids into free head groups and phosphatidic acid (PA), may regulate cellular processes through the production of lipid and lipid-derived messengers. We have genetically abrogated PLD alpha, the most prevalent isoform of PLD in plants, and the depletion of PLD alpha in Arabidopsis decreased the levels of PA and superoxide production in Arabidopsis leaf extracts. Addition of PA promoted the synthesis of superoxide in the PLD alpha-depleted plants, as measured by chemiluminescence and superoxide dismutase inhibitable, NADPH-dependent reduction of cytochrome c and nitroblue tetrazolium. The PA-enhanced generation of superoxide was associated mainly with microsomal membranes. Among various lipids tested, PA was the most effective stimulator with the optimal concentrations between 100 and 200 microM. The PA-promoted production of superoxide was observed also in leaves directly infiltrated with PA. The added PA was more effective in stimulating superoxide generation in the PLD alpha depleted leaves than in the PLD alpha-containing, wild-type leaves, suggesting that PA produced in the cell was more effective than added PA in promoting superoxide production. These data indicate that PLD plays a role in mediating superoxide production in plants through the generation of PA as a lipid messenger. PMID- 11500545 TI - Physiological and molecular biological characterization of intracellular carbonic anhydrase from the marine diatom Phaeodactylum tricornutum. AB - A single intracellular carbonic anhydrase (CA) was detected in air-grown and, at reduced levels, in high CO(2)-grown cells of the marine diatom Phaeodactylum tricornutum (UTEX 642). No external CA activity was detected irrespective of growth CO(2) conditions. Ethoxyzolamide (0.4 mM), a CA-specific inhibitor, severely inhibited high-affinity photosynthesis at low concentrations of dissolved inorganic carbon, whereas 2 mM acetazolamide had little effect on the affinity for dissolved inorganic carbon, suggesting that internal CA is crucial for the operation of a carbon concentrating mechanism in P. tricornutum. Internal CA was purified 36.7-fold of that of cell homogenates by ammonium sulfate precipitation, and two-step column chromatography on diethylaminoethyl-sephacel and p-aminomethylbenzene sulfone amide agarose. The purified CA was shown, by SDS PAGE, to comprise an electrophoretically single polypeptide of 28 kD under both reduced and nonreduced conditions. The entire sequence of the cDNA of this CA was obtained by the rapid amplification of cDNA ends method and indicated that the cDNA encodes 282 amino acids. Comparison of this putative precursor sequence with the N-terminal amino acid sequence of the purified CA indicated that it included a possible signal sequence of up to 46 amino acids at the N terminus. The mature CA was found to consist of 236 amino acids and the sequence was homologous to beta-type CAs. Even though the zinc-ligand amino acid residues were shown to be completely conserved, the amino acid residues that may constitute a CO(2)-binding site appeared to be unique among the beta-CAs so far reported. PMID- 11500546 TI - Modification of expansin transcript levels in the maize primary root at low water potentials. AB - We previously demonstrated that maintenance of cell elongation in the apical region of maize primary roots at low water potentials (psi(w)) was associated with an increase in expansin activity and extractable expansin protein. Here, we characterized the spatial pattern of expansin gene expression along the growing maize root and studied the effect of low psi(w) on expansin gene expression. Roots were divided into three segments: apical 0 to 5 mm, subapical 5 to 10 mm, and non-growing 10 to 20 mm. Of the five expansin genes expressed in control roots, two alpha-expansins (Exp1 and Exp5) and two beta-expansins (ExpB2 and ExpB8) are expressed specifically in the growing region, whereas expression of beta-expansin ExpB6 is shifted basipetally. After seedlings were transplanted to vermiculite with a psi(w) of -1.6 MPa, transcripts for Exp1, Exp5, and ExpB8 rapidly accumulated in the apical region of the root. These mRNA changes correlated with the maintenance of root elongation and increases in wall extensibility found previously. The beta-expansins ExpB2 and ExpB6 showed distinctive patterns of expression and responses to low psi(w,) indicative of distinctive functions. Inhibition of abscisic acid (ABA) accumulation at low psi(w) (by fluridone treatment) had no effect on expansin expression, except that ExpB2 transcript level showed a minor dependence on ABA. Gene-specific regulation of alpha- and beta-expansin mRNA pools likely contributes to growth alterations of the maize (Zea mays) root as it adapts to a low psi(w), but these changes do not appear to be mediated by changes in ABA content. PMID- 11500547 TI - Isolation and characterization of kinase interacting protein 1, a pollen protein that interacts with the kinase domain of PRK1, a receptor-like kinase of petunia. AB - Many receptor-like kinases have been identified in plants and have been shown by genetic or transgenic knockouts to play diverse physiological roles; however, to date, the cytosolic interacting proteins of relatively few of these kinases have been identified. We have previously identified a predominantly pollen-expressed receptor-like kinase of petunia (Petunia inflata), named PRK1, and we have shown by the antisense RNA approach that it is required for microspores to progress from the unicellular to bicellular stage. To investigate the PRK1-mediated signal transduction pathway, PRK1-K cDNA, encoding most of the cytoplasmic domain of PRK1, was used as bait in yeast (Saccharomyces cerevisiae) two-hybrid screens of pollen/pollen tube cDNA libraries of petunia. A protein named kinase interacting protein 1 (KIP1) was found to interact very strongly with PRK1-K. This interaction was greatly reduced when lysine-462 of PRK1-K, believed to be essential for kinase activity, was replaced with arginine (the resulting protein is named PRK1-K462R). The amino acid sequence of KIP1 deduced from full-length cDNA contains an EF-hand Ca(2+)-binding motif and nine predicted coiled-coil regions. The yeast two-hybrid assay and affinity chromatography showed that KIP1 interacts with itself to form a dimer or higher multimer. KIP1 is present in a single copy in the genome, and is expressed predominantly in pollen with a similar temporal pattern to PRK1. In situ hybridization showed that PRK1 and KIP1 transcripts were localized in the cytoplasm of pollen. PRK1-K phosphorylated KIP1 NT (amino acids 1--716), whereas PRK1-K462R only weakly phosphorylated KIP1-NT in vitro. PMID- 11500548 TI - Characterization of rice anthranilate synthase alpha-subunit genes OASA1 and OASA2. Tryptophan accumulation in transgenic rice expressing a feedback insensitive mutant of OASA1. AB - Anthranilate synthase (AS) is a key enzyme in the synthesis of tryptophan (Trp), indole-3-acetic acid, and indole alkaloids. Two genes, OASA1 and OASA2, encoding AS alpha-subunits were isolated from a monocotyledonous plant, rice (Oryza sativa cv Nipponbare), and were characterized. A phylogenetic tree of AS alpha-subunits from various species revealed a close evolutionary relationship among OASA1 and Arabidopsis ASA2, Ruta graveolens AS alpha 2, and tobacco ASA2, whereas OASA2, Arabidopsis ASA1, and R. graveolens AS alpha 1 were more distantly related. OASA1 is expressed in all tissues tested, but the amount of its mRNA was greater in panicles than in leaves and roots. The abundance of OASA2 transcripts is similar among tissues and greater than that of OASA1 transcripts; furthermore, OASA2 expression was induced by a chitin heptamer, a potent elicitor, suggesting that OASA2 participates in secondary metabolism. Expression of wild-type OASA1 or OASA2 transgenes did not affect the Trp content of rice calli or plants. However, transformed calli and plants expressing a mutated OASA1 gene, OASA1(D323N), that encodes a protein in which aspartate-323 is replaced with asparagine manifested up to 180- and 35-fold increases, respectively, in Trp accumulation. These transgenic calli and plants were resistant to 300 microM 5-methyl-Trp, and AS activity of the calli showed a markedly reduced sensitivity to Trp. These results show that OASA1 is important in the regulation of free Trp concentration, and that mutation of OASA1 to render the encoded protein insensitive to feedback inhibition results in accumulation of Trp at high levels. The OASA1(D323N) transgene may prove useful for the generation of crops with an increased Trp content. PMID- 11500549 TI - Plasma membrane phosphatidylinositol 4,5-bisphosphate levels decrease with time in culture. AB - During the stationary phase of growth, after 7 to 12 d in culture, the levels of phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)) decreased by 75% in plasma membranes of the red alga Galdieria sulphuraria. Concomitant with the decrease in PtdInsP(2) levels in plasma membranes, there was an increase in PtdInsP(2) in microsomes, suggesting that the levels of plasma membrane PtdInsP(2) are regulated differentially. The decline of PtdInsP(2) in plasma membranes was accompanied by a 70% decrease in the specific activity of PtdInsP kinase and by reduced levels of protein cross-reacting with antisera against a conserved PtdInsP kinase domain. Upon osmotic stimulation, the loss of PtdInsP(2)from the plasma membrane increased from 10% in 7-d-old cells to 60% in 12-d-old cells, although the levels of inositol 1,4,5-trisphosphate (InsP(3)) produced in whole cells were roughly equal at both times. When cells with low plasma membrane PtdInsP(2) levels were osmotically stimulated, a mild osmotic stress (12.5 mM KCl) activated PtdInsP kinase prior to InsP(3) production, whereas in cells with high plasma membrane PtdInsP(2), more severe stress (250 mM KCl) was required to induce an increase in PtdInsP kinase activity. The differential regulation of a plasma membrane signaling pool of PtdInsP(2) is discussed with regard to the implications for understanding the responsive state of cells. PMID- 11500550 TI - Enhanced copper tolerance in Silene vulgaris (Moench) Garcke populations from copper mines is associated with increased transcript levels of a 2b-type metallothionein gene. AB - Silene vulgaris (Moench) Garcke has evolved populations with extremely high levels of copper tolerance. To evaluate the role of metallothioneins (MTs) in copper tolerance in S. vulgaris, we screened a cDNA library derived from a highly copper-tolerant population using Arabidopsis-based MT probes and identified an MT2b-like gene. When expressed in yeast, this gene, SvMT2b, restored cadmium and copper tolerance in different hypersensitive strains. Northern-blot analysis and quantitative reverse transcriptase-PCR showed that plants from the copper tolerant S. vulgaris populations had significantly higher transcript levels of SvMT2b than plants from the copper-sensitive populations, both in roots and shoots and with and without copper exposure. Southern-blot analysis suggested that the higher expression of the latter allele was caused by gene amplification. Segregating families of crosses between copper-sensitive and copper-tolerant plants exhibited a 1 to 3 segregation for SvMT2b expression. Allele-specific PCR showed that low-expression F(3) plants were homozygous for the allele inherited from the copper-sensitive parent, whereas high-expression plants possessed at least one allele from the tolerant parent. SvMT2b expression did not cosegregate with copper tolerance in crosses between sensitive and tolerant plants. However, a significant cosegregation with copper tolerance did occur in families derived from crosses between moderately tolerant F(3) plants with different SvMT2b genotypes. Thus, overexpression of SvMT2b conferred copper tolerance although only within the genetic background of a copper tolerant plant. PMID- 11500551 TI - T-DNA-associated duplication/translocations in Arabidopsis. Implications for mutant analysis and functional genomics. AB - T-DNA insertion mutants have become a valuable resource for studies of gene function in Arabidopsis. In the course of both forward and reverse genetic projects, we have identified novel interchromosomal rearrangements in two Arabidopsis T-DNA insertion lines. Both rearrangements were unilateral translocations associated with the left borders of T-DNA inserts that exhibited normal Mendelian segregation. In one study, we characterized the embryo defective88 mutation. Although emb88 had been mapped to chromosome I, molecular analysis of DNA adjacent to the T-DNA left border revealed sequence from chromosome V. Simple sequence length polymorphism mapping of the T-DNA insertion demonstrated that a >40-kbp region of chromosome V had inserted with the T-DNA into the emb88 locus on chromosome I. A similar scenario was observed with a prospective T-DNA knockout allele of the LIGHT-REGULATED RECEPTOR PROTEIN KINASE (LRRPK) gene. Whereas wild-type LRRPK is on lower chromosome IV, mapping of the T DNA localized the disrupted LRRPK allele to chromosome V. In both these cases, the sequence of a single T-DNA-flanking region did not provide an accurate picture of DNA disruption because flanking sequences had duplicated and inserted, with the T-DNA, into other chromosomal locations. Our results indicate that T-DNA insertion lines--even those that exhibit straightforward genetic behavior--may contain an unexpectedly high frequency of rearrangements. Such duplication/translocations can interfere with reverse genetic analyses and provide misleading information about the molecular basis of mutant phenotypes. Simple mapping and polymerase chain reaction methods for detecting such rearrangements should be included as a standard step in T-DNA mutant analysis. PMID- 11500552 TI - A T-DNA insertion knockout of the bifunctional lysine-ketoglutarate reductase/saccharopine dehydrogenase gene elevates lysine levels in Arabidopsis seeds. AB - Plants possess both anabolic and catabolic pathways for the essential amino acid lysine (Lys). However, although the biosynthetic pathway was clearly shown to regulate Lys accumulation in plants, the functional significance of Lys catabolism has not been experimentally elucidated. To address this issue, we have isolated an Arabidopsis knockout mutant with a T-DNA inserted into exon 13 of the gene encoding Lys ketoglutarate reductase/saccharopine dehydrogenase. This bifunctional enzyme controls the first two steps of Lys catabolism. The phenotype of the LKR/SDH knockout was indistinguishable from wild-type plants under normal growth conditions, suggesting that Lys catabolism is not an essential pathway under standard growth conditions. However, mature seeds of the knockout mutant over-accumulated Lys compared with wild-type plants. This report provides the first direct evidence for the functional significance of Lys catabolism in regulating Lys accumulation in seeds. Such a knockout mutant may also provide new perspectives to improve the level of the essential amino acid Lys in plant seeds. PMID- 11500554 TI - Whole-plant gas exchange and reductive biosynthesis in white lupin. AB - Simultaneous measurements of CO(2) (CER) and O(2) (OER) exchange in roots and shoots of vegetative white lupin (Lupinus albus) were used to calculate the flow of reducing power to the synthesis of biomass that was more reduced per unit of carbon than carbohydrate. On a whole-plant basis, the diverted reductant utilization rate (DRUR which is: 4 x [CER + OER]) of shoot tissue was consistently higher than that of roots, and values obtained in the light were greater than those in the dark. An analysis of the biomass being synthesized over a 24-h period provided an estimate of whole-plant DRUR (3.5 mmol e(-) plant(-1) d(-1)), which was similar to that measured by gas exchange (3.2 mmol e(-) plant( 1) d(-1)). Given that nitrate reduction to ammonia makes up about 74% of whole plant DRUR, root nitrate reduction in white lupin was estimated to account for less than 43% of whole-plant nitrate reduction. The approach developed here should offer a powerful tool for the noninvasive study of metabolic regulation in intact plants or plant organs. PMID- 11500553 TI - Purification and biochemical properties of phytochromobilin synthase from etiolated oat seedlings. AB - Plant phytochromes are dependent on the covalent attachment of the linear tetrapyrrole chromophore phytochromobilin (P Phi B) for photoactivity. In planta, biliverdin IX alpha (BV) is reduced by the plastid-localized, ferredoxin (Fd) dependent enzyme P Phi B synthase to yield 3Z-P Phi B. Here, we describe the >50,000-fold purification of P Phi B synthase from etioplasts from dark-grown oat (Avena sativa L. cv Garry) seedlings using traditional column chromatography and preparative electrophoresis. Thus, P Phi B synthase is a very low abundance enzyme with a robust turnover rate. We estimate the turnover rate to be >100 s( 1), which is similar to that of mammalian NAD(P)H-dependent BV reductase. Oat P Phi B synthase is a monomer with a subunit mass of 29 kD. However, two distinct charged forms of the enzymes were identified by native isoelectric focusing. The ability of P Phi B synthase to reduce BV is dependent on reduced 2Fe-2S Fds. A K(m) for spinach (Spinacea oleracea) Fd was determined to be 3 to 4 microM. P Phi B synthase has a high affinity for its bilin substrate, with a sub-micromolar K(m) for BV. PMID- 11500555 TI - Rapid low temperature-induced stomatal closure occurs in cold-tolerant Commelina communis leaves but not in cold-sensitive tobacco leaves, via a mechanism that involves apoplastic calcium but not abscisic acid. AB - Commelina communis stomata closed within 1 h of transferring intact plants from 27 degrees C to 7 degrees C, whereas tobacco (Nicotiana rustica) stomata did not until the leaves wilted. Abscisic acid (ABA) did not mediate cold-induced C. communis stomatal closure: At low temperatures, bulk leaf ABA did not increase; ABA did not preferentially accumulate in the epidermis; its flux into detached leaves was lower; its release from isolated epidermis was not greater; and stomata in epidermal strips were less sensitive to exogenous ABA. Stomata of both species in epidermal strips on large volumes of cold KCl failed to close unless calcium was supplied. Therefore, the following cannot be triggers for cold induced stomatal closure in C. communis: direct effects of temperature on guard or epidermal cells, long-distance signals, and effects of temperature on photosynthesis. Low temperature increased stomatal sensitivity to external CaCl(2) by 50% in C. communis but only by 20% in tobacco. C. communis stomata were 300- to 1,000-fold more sensitive to calcium at low temperature than tobacco stomata, but tobacco epidermis only released 13.6-fold more calcium into bathing solutions than C. communis. Stomata in C. communis epidermis incubated on ever decreasing volumes of cold calcium-free KCl closed on the lowest volume (0.2 cm(3)) because the epidermal apoplast contained enough calcium to mediate closure if this was not over diluted. We propose that the basis of cold-induced stomatal closure exhibited by intact C. communis leaves is increased apoplastic calcium uptake by guard cells. Such responses do not occur in chill-sensitive tobacco leaves. PMID- 11500556 TI - Harpin induces activation of the Arabidopsis mitogen-activated protein kinases AtMPK4 and AtMPK6. AB - Mitogen-activated protein kinases (MAPKs) are key enzymes that mediate adaptive responses to various abiotic and biotic stresses, including pathogen challenge. The proteinaceous bacterial elicitor harpin (secreted by Pseudomonas syringae pv syringae) activates two MAPKs in suspension cultures of Arabidopsis var. Landsberg erecta. In this study, we show that harpin and exogenous hydrogen peroxide (H(2)O(2)) activate myelin basic protein kinases in Arabidopsis leaves. Using anti-AtMPK4 and anti-AtMPK6 antibodies, we identify the harpin-activated MAPKs in both leaves and suspension cultures as AtMPK4 and AtMPK6, and show that H(2)O(2), generated by Arabidopsis cells in response to challenge with harpin, activates only AtMPK6. However, treatments with catalase, which removes H(2)O(2), or diphenylene iodonium, which inhibits superoxide and H(2)O(2) production, do not inhibit harpin-induced activation of AtMPK4 or AtMPK6. In addition, activation of AtMPK4 but not AtMPK6 is inhibited by the MAPK kinase inhibitor PD98059. Neither harpin nor H(2)O(2) has any effect on AtMPK4 or AtMPK6 gene expression. In addition, the expression of AtMEKK1, AtMEK1, or AtMKK2, previously shown to be potential functional partners of AtMPK4, were not affected by either harpin or H(2)O(2) treatments. These data suggest that harpin activates several signaling pathways, one leading to stimulation of the oxidative burst and others leading to the activation of AtMPK4 or AtMPK6. PMID- 11500557 TI - Cold acclimation-induced WAP27 localized in endoplasmic reticulum in cortical parenchyma cells of mulberry tree was homologous to group 3 late-embryogenesis abundant proteins. AB - We have shown that two 27-kD proteins, designated as WAP27A and WAP27B, were abundantly accumulated in endoplasmic reticulum-enriched fractions isolated from cortical parenchyma cells of mulberry tree (Morus bombycis Koidz.) during winter (N. Ukaji, C. Kuwabara, D. Takezawa, K. Arakawa, S. Yoshida, S. Fujikawa [1999] Plant Physiol 120: 480--489). In the present study, cDNA clones encoding WAP27A and WAP27B were isolated and characterized. The deduced amino acid sequences of WAP27A and WAP27B cDNAs had 12 repeats of an 11-mer amino acid motif that was the common feature of group 3 late-embryogenesis-abundant proteins. Under field conditions, transcripts of WAP27 genes were initially detected in mid-October, reached maximum level from mid-November to mid-December, and then gradually decreased. The transcript levels of WAP27 genes in cortical parenchyma cells harvested in October was drastically induced by cold treatment within a few days, whereas those in cortical parenchyma cells harvested in August were low even by cold treatment for 3 weeks. Immunocytochemical analysis by electron microscopy confirmed that WAP27 was localized specifically in vesicular-form ER and also localized in dehydration-induced multiplex lamellae-form ER. The role of WAP27 in the ER is discussed in relation to acquisition of freezing tolerance of cortical parenchyma cells in mulberry tree during winter. PMID- 11500558 TI - A novel phytase with sequence similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings. AB - Phytic acid (myo-inositol hexakisphosphate) is the major storage form of phosphorus in plant seeds. During germination, stored reserves are used as a source of nutrients by the plant seedling. Phytic acid is degraded by the activity of phytases to yield inositol and free phosphate. Due to the lack of phytases in the non-ruminant digestive tract, monogastric animals cannot utilize dietary phytic acid and it is excreted into manure. High phytic acid content in manure results in elevated phosphorus levels in soil and water and accompanying environmental concerns. The use of phytases to degrade seed phytic acid has potential for reducing the negative environmental impact of livestock production. A phytase was purified to electrophoretic homogeneity from cotyledons of germinated soybeans (Glycine max L. Merr.). Peptide sequence data generated from the purified enzyme facilitated the cloning of the phytase sequence (GmPhy) employing a polymerase chain reaction strategy. The introduction of GmPhy into soybean tissue culture resulted in increased phytase activity in transformed cells, which confirmed the identity of the phytase gene. It is surprising that the soybean phytase was unrelated to previously characterized microbial or maize (Zea mays) phytases, which were classified as histidine acid phosphatases. The soybean phytase sequence exhibited a high degree of similarity to purple acid phosphatases, a class of metallophosphoesterases. PMID- 11500559 TI - Origins of phytochrome-modulated Lhcb mRNA expression in seed plants. AB - The levels of Lhcb mRNA in higher plants are regulated by phytochrome, cryptochrome, and an endogenous circadian oscillator. To determine whether similar regulatory mechanisms operate in the ancient gymnosperm Ginkgo biloba, we measured Lhcb mRNA levels in seedlings in response to different light conditions. Removal of a diurnally oscillating light stimulus caused dampening of maximal Lhcb mRNA accumulation levels, with little change in periodicity. Although low fluence pulses of both red and blue light given to etiolated seedlings caused maximal accumulation of Lhcb mRNAs characteristic of the phasic/circadian response seen in flowering plants, the additional initial acute response seen in flowering plants was absent. The induction of Lhcb gene expression in both cases was at least partially reversible by far-red light, and appeared biphasic over a range of red fluences. Together, these data indicate that Lhcb genes in G. biloba appear to be regulated in a manner similar to that of flowering plants, whereas signaling and attenuation of mRNA levels through the photoreceptor systems and circadian clock show features distinct from those characterized to date. The implications for these findings are discussed in light of the evolution of circadian clock input signaling. PMID- 11500560 TI - The expression of alternative oxidase and uncoupling protein during fruit ripening in mango. AB - The expression of alternative oxidase (Aox) and uncoupling proteins (Ucp) was investigated during ripening in mango (Mangifera indica) and compared with the expression of peroxisomal thiolase, a previously described ripening marker in mango. The multigene family for the Aox in mango was expressed differentially during ripening. Abundance of Aox message and protein both peaked at the ripe stage. Expression of the single gene for the Ucp peaked at the turning stage and the protein abundance peaked at the ripe stage. Proteins of the cytochrome chain peaked at the mature stage of ripening. The pattern of protein accumulation suggested that increases in cytochrome chain components played an important role in facilitating the climacteric burst of respiration and that the Aox and Ucp may play a role in post-climacteric senescent processes. Because both message and protein for the Aox and Ucp increased in a similar pattern, it suggests that their expression is not controlled in a reciprocal manner but may be active simultaneously. PMID- 11500561 TI - Sequence architecture downstream of the initiator codon enhances gene expression and protein stability in plants. AB - Nucleotide positions conserved on the 3' side of the initiator codon ATG and the corresponding N-terminal amino acid residues in a number of highly abundant plant proteins were identified by computational analysis of a dataset of highly expressed plant genes. The reporter genes uidA and gfp were modified to introduce these features. Insertion of GCT TCC TCC after the initiator codon ATG augmented expression for both the reporter genes. The insertion of each successive codon improved the expression of beta-glucuronidase (GUS) in an incremental fashion in transient transformation of tobacco (Nicotiana tabacum) leaves. The insertion of alanine-serine (Ser)-Ser resulted in about a 2-fold increase in the stability of GUS. However, this did not account for the 30- to 40-fold increase in GUS activity between the constructs coding for methionine-alanine-Ser-Ser-GUS and the native enzyme. Substitution of the codon for Ser at the third amino acid residue with synonymous codons reduced GUS expression. The results suggest a role for the conserved nucleotides in the +4 to +11 region in augmenting posttranscriptional events in the expression of genes in plants. PMID- 11500562 TI - Expression of 35S::Pto globally activates defense-related genes in tomato plants. AB - The tomato (Lycopersicon esculentum) resistance gene Pto confers resistance to the bacterial pathogen Pseudomonas syringae pv tomato carrying the avirulent gene avrPto. Overexpressing Pto under the control of the cauliflower mosaic virus 35S promoter constitutively activates defense responses in the absence of pathogen infection and nonspecifically enhances disease resistance. To elucidate the mechanisms underlying this resistance, we isolated cDNAs corresponding to transcripts that accumulated in 35S::Pto plants. By using suppression subtractive hybridization, we isolated 82 unique cDNA clones, most of which corresponded to differentially expressed transcripts. Most of the genes examined were also induced by pathogen inoculation. Sequence analysis showed that a large number of genes encode defense-related proteins, and most had not been previously isolated from tomato. The isolated cDNAs also include those with a putative role in the oxidative burst, proteolysis, the hypersensitive response, signal transduction, and a number of genes with unknown functions. The isolation of these cDNAs of diverse functions will assist in the characterization of defense pathways activated during disease resistance. PMID- 11500563 TI - Phylogenetic relationships within cation transporter families of Arabidopsis. AB - Uptake and translocation of cationic nutrients play essential roles in physiological processes including plant growth, nutrition, signal transduction, and development. Approximately 5% of the Arabidopsis genome appears to encode membrane transport proteins. These proteins are classified in 46 unique families containing approximately 880 members. In addition, several hundred putative transporters have not yet been assigned to families. In this paper, we have analyzed the phylogenetic relationships of over 150 cation transport proteins. This analysis has focused on cation transporter gene families for which initial characterizations have been achieved for individual members, including potassium transporters and channels, sodium transporters, calcium antiporters, cyclic nucleotide-gated channels, cation diffusion facilitator proteins, natural resistance-associated macrophage proteins (NRAMP), and Zn-regulated transporter Fe-regulated transporter-like proteins. Phylogenetic trees of each family define the evolutionary relationships of the members to each other. These families contain numerous members, indicating diverse functions in vivo. Closely related isoforms and separate subfamilies exist within many of these gene families, indicating possible redundancies and specialized functions. To facilitate their further study, the PlantsT database (http://plantst.sdsc.edu) has been created that includes alignments of the analyzed cation transporters and their chromosomal locations. PMID- 11500564 TI - A novel superoxide dismutase with a high isoelectric point in higher plants. expression, regulation, and protein localization. AB - Several isoforms of superoxide dismutase (SOD) with a high isoelectric point (pI) have been identified by isoelectric focusing chromatography in protein extracts from Scots pine (Pinus sylvestris) needles. One of these isoforms, a CuZn-SOD with a pI of about 10 and thus denoted hipI-SOD, has been isolated and purified to apparent homogeneity. A cDNA encoding the hipI-SOD protein was cloned and sequenced. Northern hybridization of mRNA isolated from different organs and tissues showed that hipI-SOD has a markedly different pattern of expression compared with chloroplastic and cytosolic SOD. Furthermore, the transcript levels of hipI-SOD and cytosolic SOD were found to respond differently to mechanical wounding, treatment with oxidized glutathione, paraquat, and ozone. Immunogold electron microscopy localized the hipI-SOD in the plasma membrane of sieve cells and the Golgi apparatus of albuminous cells. Moreover, high protein density was also detected in extracellular spaces such as secondary cell wall thickenings of the xylem and sclerenchyma and in intercellular spaces of parenchyma cells. PMID- 11500565 TI - Cloning and characterization of a coronatine-regulated tyrosine aminotransferase from Arabidopsis. AB - In plants, the phytotoxin coronatine, which is an analog of the octadecanoids 12 oxo-phytodienoic acid and/or jasmonic acid, gives rise to a number of physiological responses similar to those of octadecanoids. To further elucidate the physiological role of these compounds, the differential RNA display technique was used to isolate a number of novel octadecanoid-inducible genes expressed in coronatine-treated Arabidopsis. Among these, a cDNA clone was identified that was similar to known tyrosine aminotransferases (TATs). The function was verified with the expressed recombinant protein. In Arabidopsis, the protein is present as a multimer of 98 kD, with a monomer of an apparent molecular mass of 47 kD. TAT mRNA could be induced within 2 h by various octadecanoids and by wounding of the plants. Accumulation of the TAT protein and a 5- to 7-fold increase in its enzymatic activity was observed 7 to 9 h after application of octadecanoids, coronatine, or wounding. The potential role of TAT in the defense response to herbivores and pathogens is discussed. PMID- 11500566 TI - Electrodiffusional uptake of organic cations by pea seed coats. Further evidence for poorly selective pores in the plasma membrane of seed coat parenchyma cells. AB - In developing seeds, the permeability of the plasma membrane of seed coat parenchyma cells is crucial for the supply of nutrients to the embryo. Here, we report characteristics of the transport of the organic cation choline and the basic amino acid L- histidine (His; cation at pH 5, electroneutral at pH 7) into isolated seed coats of pea (Pisum sativum). Supplied at sub-micromolar concentrations, choline(+) accumulated in the seed coat tissue 5.1 +/- 0.8-fold, His(+) 2.4 +/- 0.3-fold, and His(0) 1.3 +/- 0.2-fold. Taking into consideration that at pH 5 His influxes as a cation but effluxes as a neutral molecule, these accumulations are in reasonable agreement with (electro) diffusional uptake at the prevailing membrane potential of -55 +/- 3 mV. At a concentration of 100 mM, choline(+) and His(+), but not His(0), depolarized the membrane of the parenchyma cells and neither of the substrates was accumulated. At this concentration, the relative influx (the ratio of influx and external concentration, a measure for membrane permeability) of choline and His was approximately 10 micromol g(-1) fresh weight min(-1) M(-1), similar to that found for neutral amino acids, sucrose, glucose, and mannitol. At lower concentrations, the relative influx of choline(+) and His(+) increased because of increasingly more negative membrane potentials, giving rise to apparent saturation kinetics. It is suggested that transport of organic cations can proceed by a general, poorly selective pore in the plasma membrane of seed coat parenchyma cells. This pore is thought to be responsible for the unloading of a range of solutes that serve as nutrients for the embryo. PMID- 11500567 TI - Formation of rhamnogalacturonan II-borate dimer in pectin determines cell wall thickness of pumpkin tissue. AB - Boron (B) deficiency results in inhibition of pumpkin (Cucurbia moschata Duchesne) growth that is accompanied by swelling of the cell walls. Monomeric rhamnogalacturonan II (mRG-II) accounted for 80% to 90% of the total RG-II in B deficient walls, whereas the borate ester cross-linked RG-II dimer (dRG-II-B) accounted for more than 80% of the RG-II in control plants. The results of glycosyl residue and glycosyl linkage composition analyses of the RG-II from control and B-deficient plants were similar. Thus, B deficiency does not alter the primary structure of RG-II. The addition of (10)B-enriched boric acid to B deficient plants resulted within 5 h in the conversion of mRG-II to dRG-II-(10)B. The wall thickness of the (10)B-treated plants and control plants was similar. The formation and possible functions of a borate ester cross-linked RG-II in the cell walls are discussed. PMID- 11500568 TI - A Medicago truncatula homoglutathione synthetase is derived from glutathione synthetase by gene duplication. AB - Glutathione (GSH) and homo-GSH (hGSH) are the major low-molecular weight thiols synthesized in Medicago truncatula. Two M. truncatula cDNAs (gshs1 and gshs2) corresponding to a putative GSH synthetase (GSHS) and a putative hGSH synthetase (hGSHS) were characterized. Heterologous expression of gshs1 and gshs2 cDNAs in an Escherichia coli strain deficient in GSHS activity showed that GSHS1 and GSHS2 are a GSHS and an hGSHS, respectively. Leucine-534 and proline-535 present in hGSHS were substituted by alanines that are conserved in plant GSHS. These substitutions resulted in a strongly stimulated GSH accumulation in the transformed E. coli strain showing that these residues play a crucial role in the differential recognition of beta-alanine and glycine by hGSHS. Phylogenetic analysis of GSHS2 and GSHS1 with other eukaryotic GSHS sequences indicated that gshs2 and gshs1 are the result of a gene duplication that occurred after the divergence between Fabales, Solanales, and Brassicales. Analysis of the structure of gshs1 and gshs2 genes shows they are both present in a cluster and in the same orientation in the M. truncatula genome, suggesting that the duplication of gshs1 and gshs2 occurred via a tandem duplication. PMID- 11500569 TI - Transpiration rate. An important factor controlling the sucrose content of the guard cell apoplast of broad bean. AB - Evaporation of water from the guard cell wall concentrates apoplastic solutes. We hypothesize that this phenomenon provides two mechanisms for responding to high transpiration rates. First, apoplastic abscisic acid is concentrated in the guard cell wall. Second, by accumulating in the guard cell wall, apoplastic sucrose (Suc) provides a direct osmotic feedback to guard cells. As a means of testing this second hypothesized mechanism, the guard cell Suc contents at a higher transpiration rate (60% relative humidity [RH]) were compared with those at a lower transpiration rate (90% RH) in broad bean (Vicia faba), an apoplastic phloem loader. In control plants (constant 60% RH), the guard cell apoplast Suc content increased from 97 +/- 81 femtomol (fmol) guard cell pair(-1) to 701 +/- 142 fmol guard cell pair(-1) between daybreak and midday. This increase is equivalent to approximately 150 mM external, which is sufficient to decrease stomatal aperture size. In plants that were shifted to 90% RH before daybreak, the guard cell apoplast Suc content did not increase during the day. In accordance, in plants that were shifted to 90% RH at midday, the guard cell apoplast Suc content declined to the daybreak value. Under all conditions, the guard cell symplast Suc content increased during the photoperiod, but the guard cell symplast Suc content was higher (836 +/- 33 fmol guard cell pair(-1)) in plants that were shifted to 90% RH. These results indicate that a high transpiration rate may result in a high guard cell apoplast Suc concentration, which diminishes stomatal aperture size. PMID- 11500570 TI - Chemical composition of the Prunus laurocerasus leaf surface. Dynamic changes of the epicuticular wax film during leaf development. AB - The seasonal development of adaxial Prunus laurocerasus leaf surfaces was studied using newly developed methods for the mechanical removal of epicuticular waxes. During epidermal cell expansion, more than 50 microg leaf(-1) of alkyl acetates accumulated within 10 d, forming an epicuticular wax film approximately 30 nm thick. Then, alcohols dominated for 18 d of leaf development, before alkanes accumulated in an epicuticular wax film with steadily increasing thickness (approximately 60 nm after 60 d), accompanied by small amounts of fatty acids, aldehydes, and alkyl esters. In contrast, the intracuticular waxes stayed fairly constant during development, being dominated by triterpenoids that could not be detected in the epicuticular waxes. The accumulation rates of all cuticular components are indicative for spontaneous segregation of intra- and epicuticular fractions during diffusional transport within the cuticle. This is the first report quantifying the loss of individual compound classes (acetates and alcohols) from the epicuticular wax mixture. Experiments with isolated epicuticular films showed that neither chemical conversion within the epicuticular film nor erosion/evaporation of wax constituents could account for this effect. Instead, transport of epicuticular compounds back into the tissue seems likely. Possible ecological and physiological functions of the coordinate changes in the composition of the plant surface layers are discussed. PMID- 11500571 TI - Novel anther-specific myb genes from tobacco as putative regulators of phenylalanine ammonia-lyase expression. AB - Two cDNA clones (NtmybAS1 and NtmybAS2) encoding MYB-related proteins with strong sequence similarity to petunia (Petunia hybrida) PhMYB3 were isolated from a tobacco (Nicotiana tabacum cv Samsun) pollen cDNA library. Northern blot and in situ hybridization revealed that NtmybAS transcripts are specifically expressed in both sporophytic and gametophytic tissues of the anther including tapetum, stomium, vascular tissue, and developing pollen. Random binding site selection assays revealed that NtMYBAS1 bound to DNA sequences closely resembling consensus MYB binding sites MBSI and MBSIIG, with a higher affinity for MBSI. Transient expression analyses of the N-terminal MYB domain demonstrated the presence of functional nuclear localization signals, and full-length NtMYBAS1 was able to activate two different phenylalanine ammonia-lyase promoters (PALA and gPAL1) in tobacco leaf protoplasts. Similar analysis of truncated NtmybAS1 cDNAs identified an essential, C-terminal trans-activation domain. Further in situ hybridization analyses demonstrated strict co-expression of NtmybAS and gPAL1 in the tapetum and stomium. Despite abundant expression of NtmybAS transcripts in mature pollen, gPAL1 transcripts were not detectable in pollen. Our data demonstrate that NtMYBAS1 is a functional anther-specific transcription factor, which is likely to be a positive regulator of gPAL1 expression and phenylpropanoid synthesis in sporophytic, but not in gametophytic, tissues of the anther. PMID- 11500572 TI - 30 years later: On the problem of the relation between structure and function in the brain from a contemporary viewpoint (1996), part II. AB - In the first half of the present paper, which appeared in vol. 3, issue 4 issue of Motor Control, the authors elaborated on Bernstein's (1935/1967) idea of the ambiguity of the relationship between the central command and the peripheral effect. The authors presented maybe the strongest statement so far: It is because the means are variant that the results can be invariant. As in Bernstein's 1935 paper, this was taken as evidence that there is no one-to-one relationship between structure and function in the brain. The authors discussed the history of localization theories, pointing out that neither strong localizationism nor strong anti-localizationism (as in Lashley's equipotentiality) would help understand the relation between brain structure and function. In order to understand the nature of a "brain center" for a function, the authors argued, one has to understand the concept of "function" itself. The development of "function" does not imply that the organism learns to (re)act in a stereotyped fashion, but that a control matrix is established, with non-single-valued relationships, allowing the organism to (re)act differently every time, in accordance with the need and actual situation. At the end of the first part of the paper, the authors emphasized the importance of a new basic logic of neurophysiology. In that sense, there are certain parallels between neurophysiology in the 60s (and also today, the present editors would add) and physics around the turn of the century, when Maxwell, Boltzmann, Planck, and others, created a completely new framework for theoretical physics. PMID- 11500573 TI - Terrestrial locomotion in the black-billed magpie. I. Spatio-temporal gait characteristics. AB - Spatio-temporal gait characteristics are determined for walking, running, and out of phase hopping magpies, at velocities ranging from 0.4 to 4 m/s. Below 1 m/s, magpies walk. At higher velocities they either run or hop, the latter being preferred. Stride length and frequency during walking and running relate to speed in an identical way. It is suggested that the control of walking and running, despite the abrupt drop in duty factor and step length at the transition from walking to either running or hopping, is represented by one single intrinsic pattern. Swing phase duration is independent of speed and similar of the three gaits, pointing to a passive, mechanical control. Stride frequencies during hopping barely change with velocity, while its stride length relates to velocity in a way highly comparable to that of walking and running. Hopping step length and duty factor are indifferent from those of running. These facts, combined with the similar spatio-temporal behavior of both legs in hopping suggest fairly comparable intra-limb coordination for running and hopping, and a simple phase shift in inter-limb coordination to transform a run into a hop. PMID- 11500574 TI - Evidence of motor equivalence in a pointing task involving locomotion. AB - A pointing task was performed both while subjects stood beside and while subjects walked past targets that involved differing movement amplitudes and differing sizes. The hand kinematics were considered relative both to a fixed frame of reference in the movement environment (end effector kinematics) and to the subject's body (kinematics of the hand alone). From the former view, there were few differences between standing and walking versions of the task, indicating similarity of the kinematics of the hand. However, when the hand was considered alone, marked differences in the kinematics and spatial trajectories between standing and walking were achieved. Furthermore, kinematic analyses of the trunk showed that subjects used differing amounts of both flexion-extension and rotation movements at the waist depending on whether they were standing or walking as well as on the constraints imposed by target width and movement amplitude. The present results demonstrate the existence of motor equivalence in a combined upper and lower extremity task and that this motor equivalence is a control strategy to cope with increasing task demands. Given the complexity involved in controlling the arm, the torso, and the legs (during locomotion), the movements involved in the present tasks appear to be planned and controlled by considering the whole body as a single unit. PMID- 11500575 TI - Rambling and trembling in quiet standing. AB - The goal of this study was to explore the rambling-trembling decomposition in quiet standing. The center of pressure (COP) and the horizontal ground reaction force (F(hor)) were registered in healthy subjects standing in an upright bipedal posture on a force platform. The COP positions at the instants when F(hor) = 0 were identified (instant equilibrium points, IEP) for the anterior-posterior direction, then the COP time series, were partitioned into its components using 2 different techniques, rambling-trembling decomposition and gravity line decomposition. The two decomposition techniques provided very similar results. An unexpectedly large correlation between the trembling trajectory and the difference between COP and gravity line was found, r = 0.91 (range, 0.83 < r < 0.98). The correlation implies that the GL moves from an IEP to the subsequent IEP along a smooth trajectory that can be predicted by the spline approximation. A substantial negative cross-correlation at a zero time lag was observed between the trembling and the F(hor), -0.90 < r < -0.75. For the rambling trajectory, the coefficients of correlation with F(hor) were low, -0.33 < r < -0.05. The data support the hypothesis that during quiet standing the body sways for two reasons: the migration of the reference point (rambling) and the deviation away from that point (trembling). PMID- 11500576 TI - Parameter precuing and motor preparation. AB - A movement task was used to investigate the effects of precued variables on reaction time. The task involved rapid rotation of a hand-held manipulandum to target locations and required either pronation or supination of the forearm through short or long extent. The effects on reaction time of precues signalling target direction, extent, or a combination of direction and extent, were measured. The longest reaction times occurred when no information about direction or extent was provided in the precue (all parameters uncertain). Complete prior specification of target position produced the shortest reaction times. Specification of direction when extent was uncertain produced a significantly larger reduction in reaction time than specification of extent when direction was uncertain. Prior specification of extent also produced a small but significant reduction in reaction time relative to the condition in which direction and extent were specified in a mutually conditional manner. The results are discussed in relation to parameter precuing and motor programming, in which the direction is programmed by the pre-selection of neurons representing the muscles to be used in the task while programming of extent is represented by their level of activity during task performance. PMID- 11500577 TI - Accuracy of dynamic isometric force production: the influence of age and bimanual activation patters. AB - The purpose of this study was to investigate how children and adults control bimanual activities with the influence of kinematic variables minimized. Force and timing measures were analyzed in self-paced, isometric bimanual pinch tasks performed by 6-, 8-, 10-, 12-year-old, and adult subjects. Subjects (n = 84) performed four tasks (inphase symmetrical, antiphase reciprocal, inphase asymmetrical force-right high, inphase asymmetrical force-left high) cycling between low levels (10--30%) of maximal volitional force during three 15-s trials. Bimanual tasks requiring similar activation between the hands were performed more accurately, more quickly, and with less force and timing variability than tasks requiring different actions and/or levels of force to be produced simultaneously. Evidence of force entrainment between the hands was exhibited when force direction (increasing vs. decreasing) was similar between hands but greater relative force was required of the left hand. Lower accuracy and greater variability resulted when controlled decrement of force was required to reach the lower force targets as opposed to the upper force targets which required subjects to increase force. Subjects in the two youngest age groups exhibited lower force accuracy and greater force and timing variability relative to older children and adults. Twelve-year-old subjects approximated adults' performance in all variables. PMID- 11500578 TI - Applied research and the increasing toll of a rush to judgment. PMID- 11500579 TI - Linear lesion radiofrequency ablation in canine vagal atrial fibrillation: effects of special catheters designed for efficiency, and the critical role of lesions from the crista terminalis to the superior vena cava. AB - OBJECTIVE: To determine whether specially devised catheters could be used to place radiofrequency (RF) linear lesions quickly and efficiently for termination and/or prevention of atrial fibrillation (AF). METHODS: Two versions of 2 different types of ablating catheters were used in 12 canines with AF induced by rapid pacing during vagal stimulation. 1) Modified basket catheters in two versions, one designed to produce caudo-cranial linear lesions through extended bare electrode-splines in contact with the atrial wall; and the other designed to produce horizontal linear lesions by revolving within the atrium. Together these would form "longitude and latitude" grids in the atrium. 2) The second catheter type was 2 versions of coil electrodes with thermocouples centered under each of the large-area coil electrodes. One version of these deflectable coil electrodes was intended to produce lesions in the tricuspid valve annulus-inferior vena cava (IVC) isthmus; and along the crista terminalis from the superior vena cava (SVC) to the IVC. A different type of deflection angulation on the second version was intended to produce more horizontal lesions from the crista to the tricuspid annulus. Guidance was fluoroscopic, and by electrograms and transesophageal echo. Gross pathologic examinations followed each experiment. Prior to use in canines, all electrode configurations were tested in vitro on fresh bovine preparations suspended in saline at 37 degrees C. RESULTS: The bare spline and coil electrode catheter configurations produced discrete non-perforating non-charring lesions in the in vitro preparations. One dog died of exsanginating hemorrhage. Post mortem examination revealed the lesions to be extremely variable, ranging from no evidence of effective RF delivery to deep lesions with perforation. Seven clinical successes were achieved (6 complete), with the coil electrode catheters accounting for 5 of the 7, although the procedure times were shorter with the baskets. Critical lesions were those from the crista to the SVC. Planned trans isthmus lesions were not done, but may be needed to prevent atrial flutter not seen prior to effective AF ablation. CONCLUSIONS: Special basket and coil electrode catheters may be useful but require refinement. The finding that lesions between the crista terminalis and the SVC were critical to success may be applicable to some cases of AF in humans. PMID- 11500580 TI - Electrogram-guided radiofrequency catheter ablation of atrial tissue comparison with thermometry-guide ablation: comparison with thermometry-guide ablation. AB - OBJECTIVES: To characterize a new method for radiofrequency energy titration during ablation of atrial tissue based on reduction in electrogram amplitude. To compare this method with energy titration using electrode thermometry. BACKGROUND: Complications associated with "anatomy-based" atrial endocardial radiofrequency ablation for suppression of atrial fibrillation may be due to flawed methods of energy titration. METHODS: The effect of radiofrequency ablation on electrogram amplitude was characterized in a porcine model. A method for energy titration guided by electrogram amplitude reduction ("electrogram guided") was developed and validated prospectively. Focal (smooth and trabeculated endocardial areas) and linear (smooth endocardial areas) ablation was performed comparing energy titration guided by amplitude reduction with electrode thermometry. RESULTS: Amplitude reduction during radiofrequency application was not necessarily equal among unipolar and bipolar electrograms in the ablation region; specific patterns of reduction could be discerned, based on factors such as catheter-endocardial orientation. A criterion of >90 % reduction of unipolar and/or bipolar amplitude best predicted pathologic lesion success. Electrogram-guided focal and linear lesions in smooth areas were free of lesion complications such as endocardial charring, barotrauma, or damage to contiguous extraatrial structures. However, there was a significant incidence of insufficient lesion size, principally non-transmurality, probably due to undertitration of energy. Thermometry-guided focal and linear lesions in smooth areas were uniformly transmural but frequently evidenced complications, due to overtitration of energy. Electrogram-guided focal lesions in trabeculated areas could usually not be achieved, probably due to insufficient contact of the ablation electrode with adjacent pectinate muscles. Thermometry-guided focal lesions in trabeculated areas were smaller than electrogram-guided lesions and did not evidence complications. CONCLUSIONS: Electrogram-guided lesions in smooth endocardial areas were uncomplicated but had a significant incidence of non transmurality. Thermometry-guided lesions were uniformly transmural but were frequently complicated. PMID- 11500581 TI - Atrial fibrillation: a risk factor for increased mortality--an AVID registry analysis. AB - Emerging evidence suggests that atrial fibrillation is not a benign arrhythmia. It is associated with increased risk of death. The magnitude of association is controversial and potential causes remain unknown. Patients in the registry of the Antiarrhythmics Versus Implantable Defibrillators (AVID) Trial form the basis for this report. Baseline variables, in particular the presence or absence of a history of atrial fibrillation/flutter, were examined in relation to survival. Multivariate Cox regression was used to adjust for differences in important baseline co-variables using 27 pre-selected variables. There were 3762 subjects who were followed for an average of 773+/-420 days; 1459 (39 %) qualified with ventricular fibrillation and 2303 (61 %) with ventricular tachycardia. A history of atrial fibrillation/flutter was present in 24.4 percent. There were many differences in baseline variables between those with and those without a history of atrial fibrillation/flutter. After adjustment for baseline differences, a history of atrial fibrillation/flutter remained a significant independent predictor of mortality, (relative risk=1.20; 95 % confidence intervals=1.03-1.40; p=0.020). Antiarrhythmic drug use, other than amiodarone or sotalol, was also a significant independent predictor of mortality (relative risk 1.34; 95 % confidence intervals 1.07-1.69, p=0.011. Atrial fibrillation/flutter is a significant independent risk factor for increased mortality in patients presenting with ventricular tachyarrhythmias. This risk may have been overestimated in previous studies that could not adjust for the proarrhythmic effects of antiarrhythmic drugs other than amiodarone or sotalol. PMID- 11500582 TI - Effectiveness of bi-atrial pacing for reducing atrial fibrillation after coronary artery bypass graft surgery. AB - Atrial fibrillation (AF) is common after cardiac surgery and adds significant cost and morbidity. The use of prophylactic pacing strategies to prevent post operative AF has been controversial. We previously performed a pilot study which suggested that the combination of beta-blockers and bi-atrial pacing (BAP) may reduce AF after cardiac surgery. We prospectively randomized 118 patients to continuous BAP for up to 96 hours post-operatively versus standard therapy. All patients were treated with beta-blockers as tolerated. Patients were paced in the AAI mode at a rate of 100 pulses per minute. The primary endpoint of the study was the occurrence of sustained AF (>10 minutes). There was a significant reduction in the incidence of AF in the BAP group among patients undergoing coronary artery bypass graft surgery with or without aortic valve replacement (35 % vs. 19 % AF; OR=0.38, 95 % CI 0.15, 0.93; p <0.05). Including patients undergoing isolated aortic valve surgery (n=7), there remained a strong trend toward a reduction of AF with pacing (no atrial pacing [NAP] vs. BAP; 35 % vs. 21 % AF; OR=0.48, 95 % CI 0.21, 1.11; p=0.08). Patients age 70 or greater benefited most from pacing (NAP vs. BAP; 55 vs. 25 % AF; p<0.05), while those less than 70 years of age did not (17 vs. 18 % p=NS). There was a significant reduction in the amount of time spent in the intensive care unit among patients receiving BAP (50+/-40 vs. 37+/-25 h; p<0.05).BAP together with beta-blockade after coronary artery bypass graft surgery reduces the incidence of post-operative atrial AF. Elderly patients (age 70 or greater) appear to benefit most, and may be a group to whom this therapy should be targeted. PMID- 11500583 TI - Early reinitiation of atrial fibrillation following external electrical cardioversion in amiodarone-treated patients. AB - Early reinitiation of atrial fibrillation (ERAF) following external or internal electrical cardioversion is one of the factors determining unsuccessful electrical cardioversion. Prevention of ERAF has not been studied systematically in patients on amiodarone therapy. METHODS AND RESULTS: 22 patients had ERAF within 1 min after external electrical cardioversion of atrial fibrillation. 11 patients were on amiodarone therapy and 11 patients had no antiarrhythmic medication. The effect of atropine, post-shock atrial pacing and intravenous ajmaline on ERAF was consecutively tested in these patients. Administration of atropine before repeated defibrillation or post-shock atrial pacing prevented ERAF in 9 of the 11 patients (82%) on amiodarone therapy but in only 3 of 11 patients (27%) without amiodarone (p<0.05). In the remaining patients, intravenous ajmaline was effective in the suppression of ERAF in 5 patients without amiodarone and in 1 patient with amiodarone. The PP interval preceding the atrial premature beat reinitiating atrial fibrillation was nonsignificantly longer in amiodarone-treated patients (1127+/-419 ms) in comparison to patients without amiodarone (896+/-271ms). 27% of patients without amiodarone at the time of electrical cardioversion and 55% of patients with amiodarone remained in sinus rhythm during the follow-up of 29+/-14 and 30+/-14 months, respectively. CONCLUSIONS: ERAF in patients on amiodarone can be treated by atropine or atrial pacing to prevent bradycardia-dependent ERAF. ERAF in amiodarone-treated patients does not apparently predict late recurrence of atrial fibrillation on continued amiodarone therapy. PMID- 11500584 TI - Preterm infants with paroxysmal supraventricular tachycardia: presentation, response to therapy, and outcome. AB - OBJECTIVES: We investigated the clinical course of preterm infants with paroxysmal supraventricular tachycardia in comparison to their term counterparts. BACKGROUND: Paroxysmal supraventricular tachycardia (PSVT) is the most common arrhythmia in childhood. It is known to cause significant morbidity and rarely mortality, most commonly in infants. Yet, there is minimal information in the literature on preterm infants with PSVT. METHODS: Retrospective review of 40 infants, 26 term and 14 preterm, less than three months of age who presented with PSVT from January 1990 to January 1999. We compared the severity of first clinical presentation, in-hospital response to long-term medications, and outcome after discharge. RESULTS: Symptomatic severity on presentation was not different between preterm and term infants. Preterm infants required fewer medication trials (p=0.01) and had no recurrences after discharge in contrast with 43 % recurrence in the term infants (p<0.0001). No preterm infants had Wolff-Parkinson White syndrome (WPW) in contrast to 42 % of term infants (p=0.003). Term infants with WPW were more symptomatic (p=0.01), required more medications (p=0.004), but had a similar recurrence frequency as terms infants without WPW (p=0.95). Excluding infants with WPW, preterm infants were more severely symptomatic (p=0.02), yet no longer was there a difference in response to first medication trial (p=0.30). CONCLUSIONS: We found that preterm infants with PSVT are as severely symptomatic on presentation, require fewer medications for adequate in hospital control, and have fewer recurrences than their term counterparts. Unexpectedly, preterm infants did not present with WPW. The presence of WPW only in the term infants may account for differences in the clinical course between preterm and term infants. PMID- 11500585 TI - Interactions between pacing and arrhythmia detection algorithms in the dual chamber implantable cardioverter defibrillator. AB - Dual chamber implantable cardioverter defibrillator (ICD) combines the possibility to detect and treat ventricular and atrial arrhythmias with the possibility of modern heart stimulation techniques. Advanced pacing algorithms together with extended arrhythmia detection capabilities can give rise to new types of device-device interactions. Some of the possible interactions are illustrated by four cases documented in four models of dual chamber ICDs. Functioning of new features in dual chamber devices is influenced by the fact that the pacemaker is not a separate device but a part of the ICD system and that both are being used in a patient with arrhythmia. Programming measures are suggested to optimize use of new pacing algorithms while maintaining correct arrhythmia detection. PMID- 11500586 TI - Merits and limitations of the mode switching rate stabilization pacing algorithms in the implantable cardioverter defibrillator. AB - BACKGROUND: The 7250 Jewel AF Medtronic model of ICD is the first implantable device in which both therapies for atrial arrhythmias and pacing algorithms for atrial arrhythmia prevention are available. Feasibility of that extensive atrial arrhythmia management requires correct and synergic functioning of different algorithms to control arrhythmias. METHODS AND RESULTS: The ability of the new pacing algorithms to stabilize the atrial rate following termination of treated atrial arrhythmias was evaluated in the marker channel registration of 600 spontaneously occurring episodes in 15 patients with the Jewel AF. All patients (55+/-15 years) had structural heart disease and documented atrial and ventricular arrhythmias. Dual chamber rate stabilization pacing was present in 245 (41 %) of episodes following arrhythmia termination and was a part of the mode switching operation during which pacing was provided in the dynamic DDI mode. This algorithm could function as the atrial rate stabilization pacing only when there was a slow spontaneous atrial rhythm or in presence of atrial premature beats conducted to the ventricles with a normal AV time. In case of atrial premature beats with delayed or absent conduction to the ventricles and in case of ventricular premature beats, the algorithm stabilized the ventricular rate. The rate stabilization pacing in DDI mode during sinus rhythm following atrial arrhythmia termination was often extended in time due to the device-based definition of arrhythmia termination. This was also the case in patients, in whom the DDD mode with true atrial rate stabilization algorithm was programmed. CONCLUSIONS: The rate stabilization algorithms in the Jewel AF applied after atrial arrhythmia termination provide pacing that is not based on the timing of atrial events. Only under certain circumstances the algorithm can function as atrial rate stabilization pacing. Adjustments in availability and functioning of the rate stabilization algorithms might be of benefit for the clinical performance of pacing as part of device therapy for atrial arrhythmias. PMID- 11500587 TI - Characterization of the 16 blanking periods of the Medtronic GEM DR dual chamber defibrillators. AB - We determined the blanking periods of the Medtronic GEM DR dual chamber defibrillators by using a simulator to deliver signals mimicking arrhythmias into external devices programmed to various settings. The blanking periods for the tachycardia and bradycardia functions were measured in the atrial and ventricular channels after a paced atrial event, sensed atrial event, paced ventricular event and sensed ventricular event, adding to a total of 16 blanking periods. Our findings complement the incomplete or unclear specifications published by the manufacturer. Accurate knowledge of blanking periods is essential for the interpretation of device function. PMID- 11500588 TI - Noncontact mapping for radiofrequency ablation of complex cardiac arrhythmias. AB - Radiofrequency (RF) catheter ablation is the current treatment of choice for several cardiac arrhythmias. The conventional approach utilizing intracardiac electrograms during sinus rhythm and during tachycardia has inherent limitations, including limited two-dimensional fluoroscopic imaging and limited ability to evaluate several potential sites for ablation then go precisely to the most suitable site. Recently, a noncontact mapping system has been developed that can be used to perform single beat high resolution mapping of cardiac arrhythmias. In this report, we describe the advantage of utilizing the system in facilitating a successful outcome in 5 patients with different complex arrhythmias. PMID- 11500589 TI - Case report: severe skin burn at the site of the indifferent electrode after radiofrequency catheter ablation of typical atrial flutter. AB - Although radiofrequency (RF) catheter ablation has been shown to be an effective treatment strategy in patients with supraventricular tachycardia, RF ablation may lead to potentially serious complications. We describe a case of a 65-year old man who was transferred for catheter ablation of typical atrial flutter. 21 RF applications (mean energy: 81+/-9 watts) were applied in the temperature controlled mode (70 degrees C) between a 8-mm tip electrode and an indifferent electrode using a high-power RF generator (100 watts) until bi-directional atrial isthmus block was achieved. After the procedure, a third-degree skin burn (10x2 cm) was observed at the lateral edge of the adhesive indifferent electrode whereas the medial edge of the electrode was not fully attached to the skin surface. This case is one out of 1128 ablation procedures (0.09 %) at our institution using a high-power RF generator. The present study demonstrates a severe skin burn induced by mal-attachment of an indifferent electrode during RF ablation. Long RF energy application times, high-power settings, and heavy sedation may have contributed to the observed severity of skin damage. PMID- 11500590 TI - "Pacemaker syndrome", 70 years before the first pacemaker was implanted. PMID- 11500593 TI - Dy-EOB-DTPA: tolerance and pharmacokinetics in healthy volunteers and preliminary liver imaging in patients. AB - RATIONALE AND OBJECTIVES: To investigate the tolerance and pharmacokinetics of the new liver-specific x-ray contrast agent Dy-EOB-DTPA [(4S)-4-(4-ethoxybenzyl) 3,6,9-tris(carboxylatomethyl)-3,6,9-triazaundecanedioic acid, dysprosium (Dy) complex, disodium salt] in healthy volunteers and to obtain preliminary imaging data by abdominal spiral computed tomography (CT) in tumor patients with liver metastases. METHODS: A total of 40 healthy male volunteers received 10-minute intravenous infusions of 0.05, 0.1, 0.25, 0.375, or 0.5 mmol/kg Dy-EOB-DTPA (n = 6 per dose group) or placebo (n = 10). Blood, urine, and feces were sampled for Dy measurements by inductively coupled plasma atomic emission spectrometry (ICP AES) and for the detection of possible metabolites by high-performance liquid chromatography analysis with ICP-AES detection. Safety parameters were determined before, during, and after the study. Two patients with suspected liver metastases first received 120 mL of iopromide (300 mg iodine/mL; approximately 0.6 mmol/kg) and, 24 or 72 hours later, Dy-EOB-DTPA at a dose of 0.25 mmol/kg. Computed tomography images were obtained 50 seconds after iopromide administration and before and 90 minutes after Dy-EOB-DTPA administration. RESULTS: Dysprosium-EOB DTPA was well tolerated. At the higher doses (0.375 and 0.5 mmol/kg), there was a slight increase in side effect intensity. In general, nausea, headache, and paresthesia mainly were reported as mild to moderate adverse events. Laboratory parameters did not exceed the normal range. Electrocardiographic, vital sign, or hemodynamic parameters were not affected by contrast agent administration. The terminal half-life of elimination of Dy-EOB-DTPA was approximately 1.5 hours, total clearance was 2 to 3 mL x min(-1) x kg(-1), and the renal clearance was approximately 1.5 mL x min(-1) x kg(-1). There was a significant dose dependence for the following parameters: maximal concentration in blood, terminal half-life, mean residence time, total clearance, urinary excretion, and fecal excretion. The volume of distribution in the steady state and renal clearance were not dependent on dose. In the blood and urine, no metabolites of Dy-EOB-DTPA could be detected. In the tumor patients, CT scanning after Dy-EOB-DTPA injection increased the number of detected metastases from 27 (plain scan) to 40 (iopromide) and then to 41 (Dy-EOB-DTPA) in patient No. 1 and from 1 (plain scan and iopromide) to 3 (Dy EOB-DTPA) in patient No. 2. CONCLUSIONS: Dysprosium-EOB-DTPA was shown to be a well-tolerated liver-specific contrast agent. Its pharmacokinetic profile is characterized by a terminal half-life of approximately 1.5 hours. There are indications of saturation of liver uptake at the highest dose level of 0.5 mmol/kg. In comparison with plain scans and scans performed after iodinated contrast agent administration, Dy-EOB-DTPA seems to increase the number of detectable liver lesions. PMID- 11500594 TI - Physicochemical and biological evaluation of P792, a rapid-clearance blood-pool agent for magnetic resonance imaging. AB - RATIONALE AND OBJECTIVES: To summarize the physicochemical characterization, pharmacokinetic behavior, and biological evaluation of P792, a new monogadolinated MRI blood-pool agent. METHODS: The molecular modeling of P792 was described. The r1 relaxivity properties of P792 were measured in water and 4% human serum albumin at different magnetic fields (20, 40, 60 MHz). The stability of the gadolinium complex was assessed. The pharmacokinetic and biodistribution profiles were studied in rabbits. Renal tolerance in dehydrated rats undergoing selective intrarenal injection was evaluated. Hemodynamic safety in rats and in vitro histamine and leukotriene B4 release were also tested. RESULTS: The mean diameter of P792 is 50.5 A and the r1 relaxivity of this monogadolinium contrast agent is 29 L x mmol(-1) x s(-1) at 60 MHz. The stability of the gadolinium complex in transmetallation is excellent. The pharmacokinetic and biodistribution profiles are consistent with that of a rapid-clearance blood-pool agent: P792 is mainly excreted by glomerular filtration, and its diffusion across normal endothelium is limited. Renal and hemodynamic safety is comparable to that of the nonspecific agent gadolinium-tetraazacyclododecane tetraacetic acid. No histamine or leukotriene B4 release was found in RBL-2H3 isolated mastocytes. CONCLUSIONS: The relaxivity of P792 at clinical field is very high for a monogadolinium complex without protein binding. The pharmacokinetic and biodistribution profiles are consistent with those of a rapid-clearance blood-pool agent. Its initial safety profile is satisfactory. Experimental and clinical studies are underway to confirm the potential of P792 in MRI. PMID- 11500592 TI - Report of the NASPE/NHLBI Round Table on Future Research Directions in Atrial Fibrillation. North American Society of Pacing and Electrophysiology. PMID- 11500596 TI - Effect of dose levels on the diagnostic performance of a selenium-based digital chest system. AB - RATIONALE AND OBJECTIVES: To evaluate the effect of dose reduction on diagnostic performance by using a digital chest imaging system in which amorphous selenium serves as the x-ray detector. METHODS: Two hundred forty-seven patients were examined with the selenium system. Three sets of images were made in each patient: one set with a standard x-ray dose, one set with 55% of the standard dose, and one set with 35% of the standard dose. All 741 images were read by two radiologists. The diagnostic value of each set of images for detection of pulmonary, mediastinal, and pleural pathology was analyzed with receiver operating characteristic (ROC) methodology by using computed tomography as the reference standard. The authors also assessed the effect of sex, height, and weight of the patients on the diagnostic performance of the readers. RESULTS: The areas under the ROC curves for the detection of various abnormalities at 100%, 55%, and 35% of the standard dose for observer 1, respectively, were pulmonary opacities 0.82, 0.83, 0.84; interstitial disease 0.71, 0.70, 0.72; mediastinal disease 0.81, 0.80, 0.77; and pleural abnormalities 0.71, 0.72, 0.72. There were no statistically significant differences between the two observers. CONCLUSIONS: No statistically significant difference was found between the radiologists' performance in detecting abnormalities with standard x-ray dose images and the performance with images made with 55% and 35% of the standard dose. Sex, height, and weight had no influence on diagnostic performance. PMID- 11500597 TI - Early development of vasogenic edema in experimental cerebral fat embolism in cats: correlation with MRI and electron microscopic findings. AB - RATIONALE AND OBJECTIVES: To evaluate the magnetic resonance imaging and electron microscopic findings of the hyperacute stage of cerebral fat embolism in cats and the time needed for the development of vasogenic edema. METHODS: Magnetic resonance imaging was performed at 30 minutes (group 1, n = 9) and at 30 minutes and 1, 2, 4, and 6 hours after embolization with triolein (group 2, n = 10). As a control for group 2, the same acquisition was obtained after embolization with polyvinyl alcohol particles (group 3, n = 5). Magnetic resonance images were analyzed qualitatively and quantitatively. Electron microscopic examination was done in all cats. RESULTS: In group 1, the lesions were iso- or slightly hyperintense on T2-weighted (T2W) and diffusion-weighted (DWIs) images, hypointense on the apparent diffusion coefficient (ADC) map image, and markedly enhanced on the gadolinium-enhanced T1-weighted images (Gd-T1WIs). In group 2 at 30 minutes, the lesions were similar to those in group 1. Thereafter, the lesions became more hyperintense on T2WIs and DWIs and more hypointense on the ADC map image. The lesions were enhanced on Gd-T1WIs at all acquisition times. In group 3, the lesions showed mild hyperintensity on T2WIs at 6 hours but hypointensity on the ADC map image from 30 minutes, with a tendency toward a greater decrease over time. The lesions were not enhanced on Gd-T1WIs at any time point. Electron microscopic findings revealed discontinuity of the capillary endothelial wall, perivascular and interstitial edema, and swelling of glial and neuronal cells in groups 1 and 2. Cellular swelling and interstitial edema were more prominent in group 2. In group 3, interstitial edema was seen; however, discontinuity of the endothelial wall was absent. CONCLUSIONS: The lesions were hyperintense on T2WIs and DWIs, hypointense on the ADC map image, and enhanced on Gd-T1WIs. On electron microscopy, the lesions showed cytotoxic and vasogenic edema with disruption of the blood-brain barrier. Vasogenic edema seems to develop within 30 minutes in cerebral fat embolism in cats. PMID- 11500598 TI - Feasibility of vagus nerve stimulation-synchronized blood oxygenation level dependent functional MRI. AB - RATIONALE AND OBJECTIVES: Left cervical vagus nerve stimulation (VNS) by use of an implanted neurocybernetic prosthesis (NCP) system is effective in treating epilepsy, with open data suggesting effectiveness in depression, yet the mechanisms of action are unknown. Our objective was to develop a methodology for performing VNS-synchronized functional magnetic resonance imaging (VNS-fMRI) and then to demonstrate its feasibility for studying VNS effects. METHODS: In nine patients implanted for treatment of intractable depression, a Macintosh computer was used to detect the signal from the implanted VNS stimulator and then to synchronize fMRI image acquisition with its regular firing. RESULTS: With our VNS fMRI methodology, the blood oxygenation level-dependent response to VNS was shown in brain regions regulated by the vagus nerve: orbitofrontal and parieto occipital cortex bilaterally, left temporal cortex, the hypothalamus, and the left amygdala. CONCLUSIONS: Vagus nerve stimulation pulses from an NCP system can be detected externally to determine its firing pattern, thus allowing VNS-fMRI studies of VNS-induced brain activity. PMID- 11500599 TI - Percutaneous radiofrequency (RF) thermal ablation of rabbit tumors embedded in fat: a model for RF ablation of breast tumors. AB - RATIONALE AND OBJECTIVES: To develop an experimental tumor model for RF ablation of breast tumors surrounded by fat and to investigate the minimally invasive treatment of such tumors by saline-enhanced RF ablation monitored by ultrasound (US). METHODS: Twenty-eight VX2 tumors were implanted into the retroperitoneum of 14 rabbits and monitored by B-mode US at regular intervals of 2 to 3 days. Saline enhanced RF ablation (25-mm tip length) was performed 16 days after tumor implantation (10-minute treatment time, 28 W, 15 mL/h infusion of 0.9% NaCl, which was increased to 30 mL/h in cases of an impedance increase). Thermal lesion growth was monitored by B-mode US. Treatment was considered complete if no relapse was detectable histopathologically after a follow-up period of up to 3 weeks. RESULTS: All tumor implantations were successful, reaching sizes from 5 to 38 mm 16 days after implantation. Tumors >20 mm showed central necrosis on US. Smaller tumors without signs of necrosis required greater needle perfusion to maintain constant needle-to-tissue impedance. Local relapses occurred in 14 of 27 tumors (51.8%), all with tumors >20 mm. In 12 cases, they were already detected sonographically. In 2 cases, "islands" of vital tumor were detected only during histopathological assessment. CONCLUSIONS: VX2 tumors are implantable into hypovascular adipose tissue with a high success rate and may be used as a model for RF ablation of breast tumors. B-mode US is not suitable for guiding RF ablation of tumors embedded in fat. The method appears to be more successful with smaller tumors. PMID- 11500600 TI - Hepatocellular carcinoma: correlation between vascular endothelial growth factor level and degree of enhancement by multiphase contrast-enhanced computed tomography. AB - RATIONALE AND OBJECTIVES: To determine whether vascular endothelial growth factor (VEGF) is a histopathological factor influencing contrast enhancement of hepatocellular carcinoma (HCC) on computed tomography (CT). METHODS: Twenty-two nodular HCCs underwent multiphase helical CT and surgery. Tumor size, histological grading of differentiation, and type of hepatitis were evaluated. Tumor attenuation was graded as hyperattenuated, isoattenuated, and hypoattenuated. Immunohistochemical staining with anti-VEGF antibody was performed and scored as weak, intermediate, or strong. Spearman's rank correlation test was used. RESULTS: Tumors ranged from 1.0 to 12.0 cm (mean 5.1 cm). The degree of enhancement during the hepatic arterial phase was significantly correlated with VEGF expression. Size was negatively correlated with VEGF expression and the degree of enhancement, but histological grade and type of hepatitis were not correlated with VEGF expression, tumor size, or degree of enhancement. CONCLUSIONS: In HCC, VEGF expression is correlated with the degree of contrast enhancement during arterial-phase CT. PMID- 11500601 TI - Indirect MR arthrography of the knee: effects of low-intensity ultrasound on the diffusion rate of intravenously administered Gd-DTPA in healthy volunteers. AB - RATIONALE AND OBJECTIVES: To evaluate whether application of low-intensity ultrasound may increase the diffusion rate of intravenously administered gadopentetate dimeglumine (Gd-DTPA) and increase the amount of joint fluid on indirect magnetic resonance (MR) arthrography. METHODS: Conventional MR imaging, indirect MR arthrography, and power Doppler ultrasonography were performed before and after application of therapeutic, pulsed low-intensity ultrasound in 12 asymptomatic knees of 12 volunteers. Intra-articular diffusion of intravenously administered Gd-DTPA as measured by signal intensity differences of the intra articular joint fluid before and after ultrasound treatment was assessed. In addition, the amount of joint fluid was rated, and differences in synovial blood flow as evidenced by power Doppler ultrasonography were noted. RESULTS: All volunteers tolerated well the application of therapeutic low-intensity ultrasound. A significant increase in intra-articular diffusion of intravenously administered Gd-DTPA was noted in all knees, and an increase in joint fluid was noted in 8 of 12 knees (66.6%). Detection of power Doppler flow signal in the synovium of the suprapatellar recess was possible in one instance at posttreatment exam. CONCLUSIONS: Use of pulsed, therapeutic low-intensity ultrasound may increase the diffusion rate of intravenously administered Gd-DTPA and may induce joint effusion. PMID- 11500602 TI - Of Fibiger and fables: a cautionary tale of cockroaches and Helicobacter pylori. PMID- 11500603 TI - Unresectable hepatocellular carcinoma: the need for an individualized multidisciplinary approach. PMID- 11500604 TI - Endoscopic retrograde cholangiopancreatography and laparoscopic cholecystectomy (the relationship is strained, but they are still together for the children's sake). PMID- 11500605 TI - The use of endoscopy in patients with gastrointestinal motility problems. AB - Gastrointestinal motility disorders are a commonly encountered problem. Although some are associated with organic alterations, others are defined by their symptoms, and no anatomic or histologic organic changes are to be found. In most cases, the etiology is completely unclear. Endoscopy, with the option of obtaining biopsies for histopathologic evaluation, plays the most important role in the diagnostic workup, as it can exclude such lesions as tumors, ulcers, inflammatory processes, and diverticula and it helps to define the grade and extent of motility-associated diseases (e.g., GERD). Furthermore, endoscopic interventional procedures offer sufficient treatment of several motility-related disorders (e.g., achalasia, GERD, its associated diseases, secondary constipation). PMID- 11500606 TI - Acute liver failure. AB - Acute liver failure (ALF) is defined as hepatic encephalopathy complicating acute liver injury. The most common etiologies are acute viral hepatitis A and B, medication overdose (e.g., acetaminophen), idiosyncratic drug reactions, ingestion of other toxins (e.g., amanita mushroom poisoning), and metabolic disorders (e.g., Reye's syndrome). Despite advances in intensive care management, mortality continues to be high (40-80%) and is partly related to ALF's complications, such as cerebral edema, sepsis, hypoglycemia, gastrointestinal bleeding, and acute renal failure. Several prognostic models have been developed to determine which patients will spontaneously recover. Treatment is directed at early recognition of the complications and general supportive measures. The only proven therapy for those who are unlikely to recover is liver transplantation. Therefore, recognition of ALF is paramount, and urgent referral to a transplant center is critical to assess transplantation status. PMID- 11500607 TI - Childhood constipation: evaluation and treatment. AB - Constipation is common in children. It is estimated that between 5% and 10% of pediatric patients have constipation and/or encopresis. Constipation is the second most referred condition in pediatric gastroenterology practices, accounting for up to 25% of all visits. In this article, a practical approach is laid out for those not familiar with constipation in children. Emphasis is placed on the evaluation and management options that are available to the treating practitioner. The diagnosis of constipation requires careful history taking and interpretation. Diagnostic tests are not often needed and are reserved for those who are severely affected. The daily bowel habits of children are extremely susceptible to any changes in routine environment. Constipation and subsequent fecal retention behavior often begins soon after a child has experienced a painful evacuation. Childhood constipation can be very difficult to treat. It often requires prolonged support by physicians and parents, explanation, medical treatment, and, most important, the child's cooperation. PMID- 11500608 TI - Does short-term treatment with proton pump inhibitors cause rebound aggravation of symptoms? AB - BACKGROUND: Rebound acid hypersecretion might occur after treatment with proton pump inhibitors. This study looks for a rebound aggravation of symptoms after short-term treatment with lansoprazole. STUDY: Sixty-two patients (19 men and 43 women; mean age, 54 years; range, 32-77 years) with heartburn and regurgitation and normal upper endoscopy findings were studied in a randomized, double-blind, placebo-controlled trial with a crossover design. There were two 5-day treatment periods with lansoprazole 60 mg once daily or placebo in random order, separated by a 9-day washout period. Reflux, total, and antacid scores were calculated for each of the treatment periods. Higher scores during the placebo period in the group given lansoprazole first than in the group given placebo first indicated a rebound aggravation of symptoms. RESULTS: The mean symptom scores during the placebo period in the groups given lansoprazole first and placebo first were as follows: reflux score, 21.5 and 17.6, respectively (not significant); total score, 11.2 and 10.3, respectively (not significant); and antacid score, 8.2 and 7.2, respectively (not significant). CONCLUSIONS: There is no indication of a rebound aggravation of symptoms 12 to 14 days after a 5-day treatment with lansoprazole 60 mg once daily in patients with reflux symptoms. PMID- 11500609 TI - Unsedated small-caliber esophagogastroduodenoscopy (EGD): less expensive and less time-consuming than conventional EGD. AB - BACKGROUND: A significant portion of the costs and complications of esophagogastroduodenoscopy (EGD) are related to the use of sedation. The feasibility and tolerability of small-caliber EGD (scEGD) without sedation has been evaluated; however, there is limited data concerning times and costs associated with this procedure as compared with conventional EGD (cEGD) with sedation. STUDY: Sixteen patients underwent scEGD with the Pentax EG-1840 (outside diameter, 6 mm) without sedation. A control group of 16 patients was matched for age, sex, day, and indication of procedure. The time of procedure, time in procedure room, time in recovery room, and procedure costs were determined in both the study and control groups. After the procedure, scEGD patients completed surveys consisting of visual-analogue scales to assess tolerance and preference regarding sedation for future procedures. RESULTS: Procedure time, procedure room time, and recovery room time was 5.2, 16.3, and 9 minutes for scEGD and 13.5, 34.9, and 41.3 minutes for cEGD, respectively (p < 0.001 for all comparisons). The mean cost of scEGD, excluding physician fees, was $462.00, which was significantly lower than the $587.00 for cEGD (p < 0.001). Survey results revealed good tolerance for the unsedated procedure. CONCLUSIONS: Unsedated scEGD was well tolerated and resulted in a shorter time of procedure, less time spent in procedure room, reduced recovery room time, and lower costs as compared with cEGD with sedation. PMID- 11500610 TI - Prophylactic placement of gastrostomy feeding tubes before radiotherapy in patients with head and neck cancer: is it worthwhile? AB - BACKGROUND: After radiation treatment of head and neck cancer, placement of gastrostomy feeding tubes can be technically difficult. The practice of placing tubes before treatment is probably justified if the tube is used for more than 4 weeks and if complications are infrequent. The aim of this study was to determine the outcome of prophylactically placed gastrostomy tubes in patients with head and neck cancer at our institution from 1995 to 1999. STUDY: Data collected retrospectively from the patients' medical records included demographics, duration of tube use, and complications associated with placement. RESULTS: A total of 54 patients (40 men, 14 women) with a mean age of 68.5 years (range, 49 88 years) were studied. Thirty-one patients were treated with both surgery and radiotherapy; 17, with only radiotherapy; and 6, with chemotherapy, radiation, and surgery. The gastrostomy tube was placed before initiation of radiation treatment in 41 patients and after treatment in 13. The method of placement included pull technique (n = 41), introducer technique (n = 10), and surgical (n = 3). Four patients who had a tube placed after treatment required hospitalization for dehydration, whereas no hospitalizations were needed in the prophylactic group. The median duration of tube use was 165 days (range, 0-1,105 days). Only three patients had a complication directly related to placement. CONCLUSION: Gastrostomy tube placement before treatment is appropriate, given the median number of days required for use and limited complications associated with placement. PMID- 11500611 TI - Endoscopic therapy after laparoscopic cholecystectomy. AB - The role of endoscopic therapy after laparoscopic cholecystectomy (LC) was assessed in 62 patients referred for endoscopic retrograde cholangiopancreatography (ERCP). Patients were referred because of persistent biliary pain, jaundice, abnormal liver function tests, elevated serum amylase, abnormal ultrasound of the biliary system, or abnormal laparoscopic cholangiogram. Diagnostic imaging of the biliary system was abnormal in 47 of 62 (76%) cases. Sonographic abnormalities were seen in 33 of 57 (58%) patients: common bile duct (CBD) stones were present in 10 of 58 (17%); CBD >7 mm, in 22 of 58 (38%); and subhepatic fluid collection, in 10 of 58 (17%). Laparoscopic cholangiogram was done in nine cases, and CBD calculi were detected in eight. Magnetic resonance cholangiogram was abnormal in six of seven patients: CBD stones were seen in two cases; subhepatic fluid collection, in two; and clip-on CBD, in two. Endoscopic therapy was carried out in 46 of 62 (74%) patients: 40 of 47 (85%) had abnormal imaging (p = 0.0003); 29 of 33 (88%), abnormal sonography (p = 0.002); 10 of 10 (100%), CBD stones on sonography (p = 0.03); and 31 of 46 (67%), abnormal liver function tests before ERCP (p = 0.04). Twenty-five patients had a CBD stone extracted at ERCP. Bile leaks were treated successfully with papillotomy alone in 12 of 13 cases; a stent was necessary in 1 case. Endoscopic papillotomy was done in two patients with biliary pancreatitis, in one patient with recurrent idiopathic pancreatitis, and in five with suspected biliary dyskinesia. Surgery was necessary in four patients with a CBD transection. We conclude that most patients referred for ERCP after LC need endoscopic intervention. Abnormal imaging correlates best with the need for endoscopic therapy. PMID- 11500612 TI - The anatomical distribution of colorectal polyps at colonoscopy. AB - BACKGROUND: Colorectal carcinoma is one of the most common causes of cancer related deaths in Australia. The distribution of polyps in the colon may effect the efficacy of a screening modality. The aim of this study was to determine the age-matched anatomic location and histologic type of colorectal polyps observed at colonoscopy over a 10-year period at our endoscopy unit. STUDY: Endoscopy reports on 2,578 patients were reviewed; polyp/lesion histology and location (left, right, or both) were determined in 2,553. RESULTS: Of all polyps observed, 1,310 (51%) cases were left-sided, 510 (20%) were right-sided, and 733 (29%) were synchronous. Adenomas were present in 1,659 cases (65%); of these, 734 (44%) were left-sided only and 405 (24.5%) were right-sided only. Carcinoma was observed in 189 (7%) cases, of which 71 (37.5%) were left-sided only. There was an increased right-sided prevalence of adenoma or carcinoma with age (p = 0.0029). CONCLUSION: This was not a screening study, but it has shown that a significant number of adenomas and carcinomas lie proximal to the splenic flexure. Thus, in the absence of left-sided lesions, it is expected that examination of the colon limited to the splenic flexure would miss 23% of such lesions. The increasing right-sided prevalence of these lesions with age suggests that evaluation of the proximal bowel is particularly important in older people. PMID- 11500613 TI - Clinical characteristics of acute hemorrhagic rectal ulcer. AB - BACKGROUND: Acute hemorrhagic rectal ulcer (AHRU) occurs suddenly with painless, massive, fresh rectal bleeding in elderly, bed-ridden patients who have a serious primary disease. There have been several reports on AHRU in Japan, but in Western countries, there have been none. We examined the clinical characteristics of four cases, and we report on AHRU in Western countries. STUDY: The medical records and endoscopic findings in four patients with AHRU seen during a 3-year period were reviewed. The diagnosis of AHRU was established by clinical symptoms, endoscopic examination, and other methods. RESULTS: All of the patients were elderly and bed ridden and all had sudden onset of massive, fresh rectal bleeding without pain. Sigmoidoscopic examination of all patients showed a shallow or irregular bleeding ulcer in the lower rectum. In three cases, the clinical course was uneventful. In the other case, massive, fresh rectal bleeding recurred but transanal ligation was effective. CONCLUSIONS: When painless, massive, fresh rectal bleeding occurs in elderly and bed-ridden patients, it is important to carefully examine the lower rectum, as there is a strong possibility of AHRU. PMID- 11500614 TI - Successful living donor liver transplantation for polycystic liver in a patient with autosomal-dominant polycystic kidney disease. AB - Orthotopic liver transplantation has been recommended for patients with disabling polycystic liver disease (PCLD). Because of the shortage of cadaveric donors, living donor liver transplantation (LDLT) has been developed as an alternative. We describe the case of a woman with PCLD as an extrarenal manifestation of autosomal-dominant polycystic kidney disease (ADPKD) who was successfully palliated by LDLT. The patient was a 48-year-old woman with abdominal distention. Computed tomography showed a massively enlarged liver containing innumerable cysts, as well as bilateral kidney cysts. Hepatic and renal functions were well preserved. Genetic analysis of the family did not exclude linkage to the PKD1 locus. Two and a half years after the first examination, the patient reported severely disabling symptoms caused by the PCLD. Living donor liver transplantation was performed using a right-lobe graft. The recipient and donor were both well 8 months after the transplantation. The excised liver weighed 7.4 kg, and the histopathology revealed multiple cysts and von Meyenburg complexes in the portal areas. PMID- 11500615 TI - Chronic hepatitis C virus infection associated with primary warm-type autoimmune hemolytic anemia. AB - Autoimmune hematologic abnormalities are not well recognized in chronic hepatitis C virus infection. We demonstrate an unusual association between primary autoimmune hemolytic anemia and chronic hepatitis C virus infection. A 69-year old woman who had a history of hepatitis C virus-related liver cirrhosis was found to have deteriorating anemia with reticulocytosis when admitted to the hospital. Laboratory work revealed both positive direct and indirect Coombs' tests, and warm-type immunoglobulin G against surface antigens of red blood cells. After prednisolone therapy, her anemia improved dramatically. To our knowledge, this is the first reported case of chronic hepatitis C virus infection linked with autoimmune hemolytic anemia in its natural course, not related to prior interferon treatment. Our report suggests that isolated autoimmune hemolytic anemia may be one of the unusual hematologic manifestations of chronic hepatitis C virus infection. PMID- 11500616 TI - Rapid disappearance of hepatic adenoma after contraceptive withdrawal. AB - We present the case of a 25-year-old woman who developed a large central liver adenoma after 8 years of continuous oral contraceptive use. The first diagnosis was made by ultrasonography, after a rise in plasmatic gamma-glutamyl transpeptidase and alkaline phosphatase levels was noted. Withdrawal of the oral contraceptive was followed by shrinkage of the adenoma, with complete disappearance 9 months after the diagnosis. Hepatic adenoma (HA) still presents problems in terms of differential diagnosis and clinical management. There are reports of complete or partial regression of an HA after discontinuation of oral contraceptives, but they are poorly documented. To our knowledge, a patient with such rapid disappearance of a large HA has never been reported. PMID- 11500617 TI - Endosonographic images of low-grade lymphoma of mucosa-associated lymphoid tissue after radiotherapy. AB - Recently, radiation therapy has been reported to be effective in patients with low-grade lymphoma of mucosa-associated lymphoid tissue (MALT) of the stomach. We describe changes in endoscopic ultrasonographic (EUS) findings after radiation therapy in a patient with low-grade gastric MALT lymphoma. Endoscopic ultrasonography initially showed enlargement of regional lymph nodes and diffuse thickening of the second and third mural layers in the gastric body. Two months after the end of radiation therapy, EUS showed hypoechoic changes in the third layer that corresponded to the ulcer scar but no wall thickening or lymph node enlargement. The hypoechoic changes were thought to reflect inflammatory change. We conclude that EUS is useful for assessing the response to radiation therapy in patients with low-grade gastric MALT lymphoma. PMID- 11500618 TI - Clinical and endoscopic features of adult T-cell leukemia/lymphoma with duodenal involvement. AB - We describe three cases of adult T-cell leukemia/lymphoma (ATLL) with duodenal involvement and provide a review of the literature. The first case, a 74-year-old woman with acute subtype of ATLL, had multiple polypoid lesions from the bulbus extending into the descending portion of the duodenum. The second case, a 70-year old man with lymphoma subtype of ATLL, had a polypoid tumor in the descending portion of the duodenum and multiple protruded lesions in the small and large intestines. The third case, a 67-year-old man with lymphoma subtype of ATLL, had a flat-elevated lesion in the descending portion of the duodenum, as well as a gastric ulcerated lesion. Biopsies from these lesions showed mucosal invasion of ATLL cells in each case. All patients received combination chemotherapy, which was successful in the first and third cases, accompanied by the disappearance of gastroduodenal lesions. PMID- 11500619 TI - Recurrent peritoneal mesothelioma with long-delayed recurrence. AB - Peritoneal mesothelioma is a rare malignancy with a poor prognosis. Etiology is unclear and presentation of this disease is often atypical. There are very few cases of long-term survival in patients with peritoneal mesothelioma. We report the case of a patient with peritoneal mesothelioma of 19 years' duration. To the best of our knowledge, there have been no reports of recurrent peritoneal mesothelioma with such long-delayed recurrence and survival. Recurrent peritoneal mesothelioma in the current patient was confirmed by comparing the biopsies of omentum taken in 1979 with that taken in 1997. An extensive literature search was undertaken to look for previous reports of recurrent peritoneal mesothelioma. Aggressive and intensive chemotherapy appeared to provide a longer survival time. Our patient had paraneoplastic manifestations that included paraneoplastic hepatopathy and a wasting syndrome. Peritoneal mesothelioma is a rare malignancy with one to two cases per million in the United States. It is known to exhibit unusual paraneoplastic manifestations, including paraneoplastic hepatopathy. Treatment is often unsuccessful, with a median survival time of approximately 18 months from diagnosis. A better understanding and aggressive management of this disease appear to help prolong survival. PMID- 11500620 TI - Lactobacillus casei subsp. rhamnosus sepsis in a patient with ulcerative colitis. AB - Lactobacillus species are part of normal gastrointestinal flora and are an uncommon cause of bacteremia. Lactobacillus casei subsp. rhamnosus was repeatedly isolated in the blood cultures of a 43-year-old woman with ulcerative colitis. Lactobacillus spp. could be considered a potential opportunistic pathogen whenever there is disruption of the physiologic architecture of the intestinal mucosa, such as in abdominal surgery, endoscopic procedures, or enteric pathologies. Clinicians and clinical microbiologists need to be aware of this infective complication in patients with ulcerative colitis. PMID- 11500621 TI - Intestinal pseudoobstruction as a presenting feature of a pheochromocytoma. PMID- 11500622 TI - Patterns of response during therapy with interferon and outcome of retreatment with interferon plus ribavirin for chronic hepatitis C. PMID- 11500623 TI - Mortality associated with infliximab. PMID- 11500624 TI - Clinicopathological factors in early squamous cell carcinoma of the tongue and floor of the mouth, in Biscay (the Basque Country, Spain). AB - OBJECTIVES: A study is made of the main clinicopathological factors in squamous cell carcinoma (SCC) of the tongue and floor of the mouth in the province of Vizcaya (Biscay) (The Basque Country, Spain), and their relation to patient prognosis. DESIGN: A retrospective study was made of 40 patients with early (clinical stage I/II) SCC of the tongue and floor of the mouth. A previously designed protocol was used to record the clinical and histopathological data, which were subjected to descriptive and comparative bi- and multivariate statistical and survival analyses. RESULTS: There were 34 males and 6 females, with a mean age of 55.7 years (range 33-81). In 23 cases the SCC was located in the tongue, and in 17 cases in the floor of the mouth. The average tumor diameter was 2.6 cm; 65% of the neoplasms were ulcerated. The mean clinical course was 3.4 months, with an average tumor growth rate or velocity of 268 (Evans formula). Thirty-four patients were smokers and 33 consumed alcohol. All SCC of the floor of the mouth, and all regional recurrences, were diagnosed in smokers and drinkers. Survival at 5 years was 65%. Well differentiated SCC were diagnosed in 52.5% of cases. The average histological malignancy grade was 1.96, and was higher in males, tongue malignancies, T2 lesions, tumors with growth velocities above 200, and in recurrent neoplasms. CONCLUSIONS: Early SCC of the tongue and floor of the mouth in Vizcaya affects mainly male smokers and drinkers of alcohol under the age of 60 years. A relationship is observed between the histopathological findings (particularly invasion mode and stage) and patient prognosis. PMID- 11500625 TI - Eosinophilic granuloma as a form of inflammatory reaction. A case report. AB - Eosinophilic granuloma consists of the proliferation and/or accumulation of Langerhans cells in the bones, generally of the cranium and face, as a uni- or multifocal cystic lesion. It is considered to be a localized chronic form of Langerhans cell disease. The most frequent oral location is the posterior part of the mandible, where the bone lesion often gives rise to lesions of the overlying soft tissues. We report a case showing bilateral involvement of the upper jaw and unilateral involvement of the mandible. The eosinophilic granulomas arose in association with odontogenic periapical infectious processes, suggesting that this disorder may be a form of inflammatory response. PMID- 11500626 TI - Orofacial manifestations of histoplasmosis in HIV-positive patients: a case report. AB - Amongst the main opportunistic diseases that affect the HIV-positive patient, histoplasmosis is found. This systemic mycosis caused by the fungus Histoplasma capsulatum has the capacity to disseminate from the lung to the skin and oral mucosa. Oral lesions of histoplasmosis can be found with ulcerated or nodular aspect, being always very painful and infiltrating the mucosa. When they are present in the mouth, they strongly indicate the presence of some kind of immunosuppression. This study shows the disease's evolution in an HIV-positive patient, who presented several ulcerated lesions in the oral cavity and facial skin. The symptomatology and clinical aspects of the lesions were not specific for the disease, and due to this, the diagnosis was obtained by cytological smear and oral biopsy. The results of the exams defined the disseminated picture of the infection. The treatment plan involved the use of amphotericin B for the lesions' remission, and, following this, itraconazole was administered in the maintenance phase. PMID- 11500627 TI - Myxoma of the jaws. Report of three cases. AB - Odontogenic myxoma is a locally aggressive, uncommon benign tumour which arises from mesenchymal tissues normally present in developing teeth. The most frequent locations of odontogenic myxoma are the posterior regions of the mandible, as well as the condylar region. Since odontogenic myxomas are not associated with any specific clinical or radiological sign, a histopathological examination of the specimen is required for confirmation of the primary diagnosis. We report three cases of myxoma diagnosed during the last 18 years. Two of them were located in atypical regions of the mandible and one was located in the maxilla. Presence of a slow-growing swelling associated with expansion of the bone plates raised suspicion of a tumour in two cases, while in the third patient the myxoma was an incidental finding during radiological examination. Due to the unspecific nature of these lesions, in every case a histopathological examination of the surgical specimen was required for diagnostic confirmation. In one of the three reported cases, we shall underline the need to follow a correct diagnostic work up of all radiolucent lesions of the jaws, in order to avoid contraindicated therapeutic procedures. PMID- 11500629 TI - Peripheral ossifying fibroma. Report of a case and review of the literature. AB - Peripheral ossifying fibroma, a reactive gingival disorder known under the generic term of epulis, is widely considered to originate from the cells of the periodontal ligament. A 39-year-old woman presented with a well circumscribed, hard pedunculated exophytic tumor measuring 2 cm in size, without ulceration. The lesion had developed three months before in the interdental space, between the lower left lateral incisor and canine. No radiological signs of involvement of the alveolar ridge were observed, and the clinically manifest diastema was confirmed. The histological study of the specimen after simple resection with a cold scalpel confirmed the diagnosis of peripheral ossifying fibroma. The patient posteriorly suffered three recurrences that were respectively treated via simple resection, carbon dioxide laser exeresis and finally wide resection including the periodontal ligament, periosteum and underlying bone. There have been no further recurrences following wide resection. In addition to presenting this new case of peripheral ossifying fibroma, the present study reviews the literature, comments on the predisposing factors and treatment options, and suggests the importance of this disorder in the differential diagnosis of gingival lesions. PMID- 11500628 TI - Retinoids: application in premalignant lesions and oral cancer. AB - Squamous cell carcinoma (SCC) of the oral cavity is a multistep process, progressing through a series of discrete, irreversible and complementary alterations in genes that control cell growth, death, and differentiation. In the premalignant state, the oral mucosa progresses through various grades of epithelial dysplasia, with the potential to convert to SCC. Natural and synthetic vitamin A metabolites and analogs (retinoids) were found to suppress head and neck and lung carcinogenesis in animal models, and inhibit carcinogenesis in individuals with premalignant lesions and a high risk to develop cancer of the aerodigestive tract. Likewise, retinoids prevent the development of second primary cancers in head and neck and lung cancer patients who had been treated for the first primary. These effects are thought to result from changes in the expression of genes that regulate cell growth and differentiation. Chemopreventive strategies are designed to suppress, reverse, or prevent the formation of premalignant lesions and their subsequent progression to SCC. This is a review of the application of retinoids in the chemoprevention of head and neck cancer. PMID- 11500630 TI - Malignant tumors of the minor salivary glands. Retrospective study on 59 cases. AB - Minor salivary glands cancer account for 2-4% of head and neck cancer, 10% of all oral cavity cancer and 15-23% of all salivary glands cancer (1). There is a remarkable variation in clinical presentation, behavior and histology of those neoplasms. The incidence of malignancy is around 52-60% (2-4) Several classifications of salivary gland tumors have been presented, but for the present study we have utilized the WHO classification of 1992 (5). We present the results of a review of our experience with those malignancies, considering prognostic factors as histopathology, tumor grade, clinical staging, age and sex, primary location, regional and distant metastasis. PMID- 11500631 TI - True aneurysm of the facial artery. Report of a clinical case. AB - Aneurysms are abnormal and localized dilations of a blood vessels presenting in any artery or vein of the body. They can be classified in false aneurysms, when the have a traumatic etiology, and in true aneurysms, when the vessel wall is weakened but containing all layers. True aneurysms are not much described in literature. We present a case of a 54 year old female patient with a true aneurysm of the facial vein, its surgical treatment and its final diagnosis. The clinical and pathological aspects of these kind of lesions and differential diagnosis with other similar lesions are discussed. PMID- 11500633 TI - Anaesthesia of the right lower hemilip as a first manifestation of multiple myeloma. Presentation of a clinical case. AB - Multiple myeloma is a malignant proliferation of plasma cells. It may affect any of various bones, causing osteolytic lesions with a characteristic "punched out" radiographic appearance. The commonest symptom is bone pain. One of the most frequent locations is the mandible. Symptoms of multiple myeloma of the mandible include tumefaction, non-specific pain, tooth mobility and sometimes loss, and paraesthesia of the dental nerve. Here we report a case of multiple myeloma of the mandible which was unusual in that the presenting complaint was anaesthesia of the right lower hemilip. PMID- 11500632 TI - Cutaneous lupus erythematosus. PMID- 11500634 TI - Ameloblastic fibrosarcoma: report of a case. Immunohistochemical study and review of the literature. AB - Ameloblastic fibrosarcoma is a rare malignant odontogenic tumour characterized by a benign epithelial component within a malignant fibrous stroma. Its behaviour is relatively benign, with absence of metastatic disease, and the prognosis is reported to be good. It is a paradoxical neoplasm with "sarcomatous" morphological and immunohistochemical patterns but with a favourable clinical course. We report a new case of this tumour in a mandibular ramus of a 31-years old male patient, that was surgically excised and treated with adjuvant chemotherapy and radiotherapy. Five years later the patient is free of disease. The growth potential of ameloblastic fibrosarcoma is evaluated and compared with a related lesion, the ameloblastic fibroma. The sarcomatous mesenchymal component of ameloblastic fibrosarcoma is positive to Ki67, PCNA and p53, in front of the negativity of ameloblastic fibroma. PMID- 11500635 TI - Mucosal cysts of the maxillary sinus: a literature review. AB - The aim of this article is to review the clinical, radiographic and microscopic features, as well as the etiopathogenesis, the differential diagnosis and the treatment of mucosal cysts of the maxillary sinus. PMID- 11500636 TI - Aminopeptidase activity in human saliva. AB - In this work, different aminopeptidase activity levels in whole human saliva are described. Aminopeptidase activities were studied by measuring the rate of hydrolysis of the artificial substrates Ala-, pGlu-, Pro-, Arg-, Asp- y Cis-2 naphthylamides (fluorimetrically detected at 412 rim with excitation at 345 nm). The presence of these enzyme activities in the saliva suggests that the active levels of saliva peptides can be controlled by homeostatic mechanism similar to those that have been described in other tissues, such as plasma, the central nervous system, and immunocompetent cells. PMID- 11500637 TI - Pulsioxymetry monitorization during lower third molar extraction. A comparative study of three local anesthetics with epinephrine 1:100,000. AB - A double-blind, parallel non-randomized study is made to compare the heart rate and oxygen saturation levels in healthy patients subjected to lower third molar extractions under the effect of three different local anesthetics (4% articaine, 2% lidocaine, 2% mepivacaine) with the same epinephrine concentration (1:100,000). The patients were divided into three groups of 15 subjects each, according to the anesthetic employed. Heart rate and oxygen saturation were determined in each patient before and immediately after anesthetic injection, and again one minute after motor application to perform the ostectomy, at the start of force application to extract the tooth, on completing suture, and one week later at suture removal (the latter being taken as baseline reference). Blood pressure was also recorded, calculating the mean arterial pressure and the rate pressure product (RPP) and pressure-rate quotient (PRQ). Only heart rate showed variations during the intervention, regardless of the local anesthetic used (based on a multifactor analysis of variance and considering statistical significance for p < 0.05). It may be concluded that none of the local anesthetics studied exert important effects upon the variables analyzed in healthy patients, though significant variations may arise due to the stress and/or anxiety associated to certain moments of lower third molar extraction. PMID- 11500638 TI - Surgical management of actinic cheilitis. AB - OBJECTIVE: An analysis is made of the etiologic factors underlying actinic cheilitis and of the results obtained following surgical treatment of the disease in a series of 54 patients (32 males and 22 females). STUDY DESIGN: The case history was evaluated, along with the triggering factors, histopathological pattern and recurrence of lesions. A cold scalpel vermilionectomy was performed in 41 patients, followed by primary closure of the defect using an oral mucosal flap. The remaining 13 patients were subjected to carbon dioxide laser vermilion ablation. RESULTS: The history of the patients revealed liver disease associated to alcohol consumption in 35.2% of cases and tobacco smoking in 77.8%. As regards solar exposure, 53.1% of the men referred open-air professional activities, while 100% of the women presented important solar exposure. The most frequent clinical manifestations were bleeding and the presence of leukoplakia patches; pain was reported in only 16% of cases. Over 40% of the patients were asymptomatic. Following treatment, and after a follow-up period of at least 6 months, 90.7% of the subjects showed complete healing. Of the 5 patients reoperated upon, four healed and one evolved towards squamous cell carcinoma. CONCLUSION: Vermilionectomy is the recommended treatment for actinic cheilitis, using either a cold scalpel or carbon dioxide laser. A histological study of the lesion is indicated in all cases to secure early detection of possible malignization. The control of the possible etiologic factors is also essential. PMID- 11500639 TI - Eosinophilic granuloma of the jaws: a report of three cases. AB - Three cases of eosinophilic granuloma involving two boys and a girl are reported. Osteolytic lesions of the jaws were seen in all three patients. The two boys presented mandibular lesions, while the girl had lesions in both jaws. Patient age at the onset of the disease was 6 and 9 years in the boys and 10 years in the case of the girl. The affected zone was painful to palpation in all cases, with intra- and extraoral swelling and bone bulging that caused slight facial deformity and bleeding of the gums. Biopsies were obtained in all three cases, and the histopathological study confirmed the diagnosis. The clinical course was favorable in all three children. PMID- 11500640 TI - Cellular immune alterations in fifty-two patients with oral lichen planus. AB - OBJECTIVES: Abnormal immune mechanisms appear to play an important role in the pathogenesis of lichen planus (LP). DESIGN: A prospective clinical study was made to investigate the differences in T cell subpopulations in 52 patients with oral lichen planus (OLP) and 54 healthy controls. RESULTS: Statistically significant differences were observed among the OLP patients versus the controls: the percentages of CD3+, CD8+, DR+, CD57+ and CD8CD45RA+ cells were depressed, while elevations were recorded in the subpopulations expressing CD4+, and in the ratios CD4/CD8+ and CD8CD45RO+/CD8CD45RA+. CONCLUSIONS: These results suggest the existence of differences in the lymphocyte subpopulations between healthy subjects and patients with OLP. These findings do not coincide with the previously published observations, however; the observed cellular immune alterations thus appear to relate more to lymphocyte reaction capacity than to the actual number of cells in each population subgroup. PMID- 11500641 TI - Kikuchi-Fujimoto disease. A case report. AB - Histologically, Kikuchi-Fujimoto disease (KFD) is a necrotising, self-limiting lymphadenitis. It typically affects the lymph nodes of the neck and it is often accompanied by a high fever. The present paper documents the case of a 28-year old male carrier of KFD. From the x-ray images, the KFD clinically appeared to be a submandibular salivary gland tumor. An ultrasound study led us to believe that it was more likely a branchial cyst. Malignant lymphoma can be mistaken for KFD, thus the diagnosis must be precise and the differentiation is crucial. PMID- 11500642 TI - Recurrent infection of a complex odontoma following eruption in the mouth. AB - Odontomas are the most common odontogenic tumors, and are classified as either compound or complex lesions. Although its underlying etiology remains unclear, a number of factors appear to be involved (e.g., prior trauma). The lesions are generally diagnosed in the second decade of life, often in the context of routine dental X-ray examinations. A complex odontoma is described in a 22-year-old woman referring discomfort due to overinfection following its aperture into the oral cavity in the distal alveolar region of 2.6. Initial treatment consisted of antibiotics and antiinflammatory drugs. Computed axial tomography revealed a solid, irregular mass in the distal zone of the left maxilla, imprinting upon the region of the maxillary sinus on the same side. Surgical resection was performed, raising the alveolar mucosa and upper left vestibular fundus. The surgical piece contained the amorphous adenoma mass and the tooth 2.7. PMID- 11500643 TI - Oral lesions due to child abuse. Identification of the problem and role of the professional. AB - Violence in the family setting is a serious social problem of obliged concern to all health professionals. The victims are mainly children, women and the elderly. Due to the defenselessness characterizing childhood and the requirements of Spanish law, the dentist must be aware of the magnitude of the situation in order to act in favor of the affected child. The problems encountered in this aspect comprise physical abuse, sexual abuse, physical neglect, emotional abuse and emotional neglect. Any of these problems can have irreversible consequences for the psychological and physical development of the child. A review is made of the characteristic orofacial lesions that should alert dental professionals to the possible existence of abuse, with a discussion of the approach to be adopted within the context of current legislation. PMID- 11500644 TI - Characteristics and dental treatment of partial trisomy 9. AB - A description is made of the dental characteristics and treatment of a patient with partial trisomy 9. Due to the deep mental retardation and motor deficiency of the patient involved, treatment (extractions, caries restorations and periodontal treatment) is conducted in hospital and under general anesthesia. Trisomy 9 has only rarely been described in literature, and the present study provides one of the first descriptions of both the dental characteristics of these patients and the treatment of their oral pathology. PMID- 11500645 TI - Prevalence of biopsied oral lesions in a service of oral surgery. AB - INTRODUCTION: A retrospective study is made of 205 case histories of patients subjected to biopsy of an oral cavity lesion. PATIENTS, MATERIAL AND METHODS: The study covered an 18-month period in which 2960 first visits were documented, including 205 oral lesions amenable to histological evaluation. Patient age and sex was evaluated, along with the clinical manifestations and location of the lesions, the biopsy technique used, the size of the tissue fragment referred for histological study, the reason for consultation (related to the lesion or otherwise), and the histological results obtained. RESULTS: The most frequently biopsied lesions were cysts, including particularly root cysts (19.5%). Periapical granulomas (18.1%) and fibromas (15.7%) were also common in our series. The histological findings were varied, with a total of 35 different diagnoses. Two of the 205 biopsied oral lesions were shown to be squamous cell carcinomas (1%). CONCLUSIONS: The frequency of oral lesions susceptible to histopathological study was similar to the figures reported elsewhere in the literature. PMID- 11500646 TI - Ask us: Are there predictors for root resorption? PMID- 11500647 TI - No exemptions from lessons in humility. PMID- 11500648 TI - The necessity of thinking. PMID- 11500649 TI - California proposition may help patients in search of better oral health. PMID- 11500650 TI - The importance of incisor positioning in the esthetic smile: the smile arc. AB - The smile arc is defined as the relationship of the curvature of the incisal edges of the maxillary incisors and canines to the curvature of the lower lip in the posed smile. The ideal smile arc has the maxillary incisal edge curvature parallel to the curvature of the lower lip. Evaluation of anterior smile esthetics must include both static and dynamic evaluations of profile, frontal, and 45 degrees views to optimize both dental and facial appearance in orthodontic planning and treatment. This article presents the concept of the smile arc and how it relates to orthodontics-from the recognition of its importance, to its impact on orthodontic treatment planning, to how procedures and mechanics are adapted to optimize the appearance of the smile. Three cases are used to illustrate how treatment is directed, emphasizing how facial and smile goal setting go hand in hand. PMID- 11500651 TI - Malocclusion as an etiologic factor in periodontal disease: a retrospective essay. PMID- 11500652 TI - Open gingival embrasures after orthodontic treatment in adults: prevalence and etiology. AB - The purposes of this study were to determine the prevalence of posttreatment open gingival embrasures in adult orthodontic patients and to examine the association of pretreatment maxillary incisor malalignment, posttreatment alveolar bone height, interproximal contact position, root angulation, crown shape, and embrasure area with open gingival embrasures. Posttreatment intraoral photographs of 337 adult orthodontic patients were evaluated to determine the prevalence of open gingival embrasures. A subsample of 119 patients was identified for measurement and divided into 2 groups: normal gingival embrasures and open gingival embrasures. Digital images of the pretreatment maxillary models and posttreatment maxillary central incisor periapical radiographs were made to measure the pretreatment and posttreatment variables. The prevalence of posttreatment open gingival embrasures in adult orthodontic patients was 38%. Pretreatment maxillary central incisor rotation and overlap were not statistically associated with posttreatment open gingival embrasures. A posttreatment alveolar bone-interproximal contact distance greater than 5.5 mm was associated with open gingival embrasures. Short and more incisally positioned posttreatment interproximal contacts were associated with open gingival embrasures. Open gingival embrasures were found to have more divergent root angulations and more divergent or triangular-shaped crown forms than normal gingival embrasures. Embrasure areas larger than 5.09 mm(2) were also correlated with open gingival embrasures. Increased alveolar bone-interproximal contact distance and increased root angulation demonstrated the greatest increase in the odds of an association with an open gingival embrasure. This investigation indicates that open gingival embrasures are common in adults who have undergone orthodontic treatment and that posttreatment variables are significant factors in open gingival embrasures. PMID- 11500653 TI - Predicting upper lip response to 4-piece maxillary LeFort I osteotomy. AB - The purpose of this retrospective study was to understand and predict the multidimensional changes in upper lip morphologic features after segmental (4 piece) maxillary Le Fort I advancement/impaction with VY closure and alar base cinch sutures. The study evaluated longitudinal lateral cephalograms of 57 patients (42 women, 15 men) 27.5 +/- 11.2 years of age before surgery. Lateral cephalograms with teeth in occlusion and lips in repose were taken 2 weeks before surgery and at least 6 months after the operation. Mean postsurgical duration was 15.5 months. The upper lip predictably moved anteriorly in a graduated fashion, from 50% (subnasale) to 90% (labrale superius) the amount of the underlying osseous anterior movement, and showed a slight lengthening (0.73 +/- 1.9 mm) from subnasale to upper lip stomion. The upper lip surface contour was also straightened as a result of the surgical movement. Multiple regression models showed that the anterior changes in the landmarks prosthion and facial surface of the upper incisor were the most important variables in predicting upper lip response. The prediction equations for horizontal movements explained 86% to 94% of the variation, with errors of the estimates that range between 1.27 mm and 1.65 mm. The models, when applied to an independent validation sample of 14 subjects, explained between 86% and 94% of the total variation. The conclusion is that upper lip response after 4-piece Le Fort I advancement/impaction (VY closure and alar base cinch suture) can be accurately predicted. PMID- 11500654 TI - Stability of skeletal Class II correction with 2 surgical techniques: the sagittal split ramus osteotomy and the total mandibular subapical alveolar osteotomy. AB - Combined orthodontic and surgical treatment of severe Class II dentoskeletal deformities with the use of the bilateral sagittal split ramus osteotomy is a routine procedure in orthodontic practices. However, an alternative surgical technique, the total mandibular subapical alveolar osteotomy, could be used for the same purpose. The aim of this investigation was to compare the stability of the sagittal split ramus osteotomy with the total mandibular subapical alveolar osteotomy in the correction of dentoskeletal Class II malocclusions. Forty patients that exhibited Class II dentoskeletal relationships were included in the study. Twenty of these patients had mandibular advancement with the sagittal split ramus osteotomy; the remaining 20 patients had advancement of the whole lower alveolar segment with the total mandibular subapical alveolar osteotomy. The cephalograms studied were taken before the surgical procedure (T1 = 4 weeks before operation), immediately after the procedure (T2 = 10 days after surgery), and 1 year later (T3). The statistical analysis used to assess the results between and within the groups over the different time periods was the analysis of variance. The regression analysis was used to test the interdependence of soft tissue response to hard tissue movement. The results of this study show that both procedures are equally stable when correcting Class II malocclusions. This was proved by the stability of the correction of overjet, B point, and incisor mandibular plane angle. There were no statistically significant differences between or within the groups in the position of these landmarks over time. There was a statistically significant change in the position of pogonion from T1 to T2 (P <.0028) between the groups, although at T3 this difference was not significant (P <.05). There were no significant changes in face height either within or between the groups over time. The hard/soft tissue interactions for the total mandibular subapical alveolar osteotomy were as follows: The lower lip advanced 60% to the incisor movement; soft tissue B' point responded with a 130% advancement in relation to the change in its hard tissue counterpart. Soft tissue pogonion advanced 90% in relation to the hard tissue landmark. The data suggest that the total mandibular alveolar osteotomy is the treatment of choice for the correction of severe dentoalveolar retrusive Class II malocclusion for which alteration of the mentolabial sulcus is desirable. PMID- 11500655 TI - Maxillary sagittal and vertical displacement induced by surgically assisted rapid palatal expansion. AB - The purpose of this study was to investigate the sagittal and vertical effects on the maxilla induced by surgically assisted rapid palatal expansion. Twenty patients (average age, 25.6 years) who required a surgically assisted rapid palatal expansion procedure were available for this study. Each patient was banded with a Haas-type palatal expander, maxillary surgery was performed, and the expander was activated. Presurgical and postexpansion lateral cephalograms were taken for each patient. The measurements of SNA, Frankfort horizontal-nasion A point, sella-nasion-palatal plane, A-point-nasion-perpendicular (in millimeters), 1-nasion-A-point, 1-sella-nasion, 1-nasion-perpendicular (in millimeters) were made on each cephalogram, and the presurgical and postexpansion tracings were superimposed on the cranial base to determine the changes in the anterior nasal spine and posterior nasal spine positions. Results showed that from the presurgical cephalogram to the postexpansion cephalogram SNA, Frankfort horizontal-nasion-A point and A-point-nasion-perpendicular had a mean increase of 0.60 degrees (P <.05), 0.65 degrees (P <.05), and 0.55 mm (P <.05), respectively; 1-nasion-A-point decreased 2.18 degrees (P <.05) and 1-sella-nasion decreased 1.53 degrees (P <.05). No significant maxillary vertical displacement was noted from the first cephalogram to the second. In conclusion, surgically assisted RPE did not significantly affect the maxilla vertically; however, it did induce a slight forward movement of the maxilla and a slight retroclination of the maxillary incisors sagittally (P <.05). PMID- 11500656 TI - Facial symmetry after distraction osteogenesis and orthodontic therapy. AB - The purpose of this study was to document changes in the facial symmetry of patients with severe hypoplastic mandibles treated with distraction osteogenesis and orthodontic therapy. Ten patients (5 males and 5 females), ages 7 to 22 years, underwent unilateral mandibular distraction osteogenesis with extraoral distracting devices. Orthodontic therapy was initiated after the completion of mandibular lengthening. The mean follow-up period was 25.5 months. Posteroanterior cephalograms were traced before distraction, 6 months after distraction, and at a later follow-up appointment. Displacement of the chin point (in millimeters) and the midpoint of the mandibular incisors, the piriform angle, the intergonial angle, the occlusal cant to the horizontal line (in degrees), the ratio of linear measurements (in millimeters) Co-Gn, Co-Go, and Go-Gn of the affected to the nonaffected side, CoA'-Mid to CoA-Mid, and GoA'-Mid to GoA-Mid were compared. At the latest follow-up appointment, patients defined as having a successful result had a mean occlusal cant of 1.25 degrees, an intergonial angle of 2.13 degrees, and a piriform angle of 2.63 degrees. The mean displacement of the chin point to the midline was 1.5 mm and that of the mandibular central incisors to the midvertical line was 1.38 mm. The results indicated improvement in all patients. PMID- 11500657 TI - Pharyngeal airway space changes after counterclockwise rotation of the maxillomandibular complex. AB - The purpose of this study was to evaluate the effects of double-jaw surgery with counterclockwise rotation of the maxillomandibular complex on the pharyngeal airway space and velopharyngeal anatomy in patients with high occlusal plane facial morphology. Fifty patients (22 men, 28 women) with high occlusal plane facial morphology underwent double-jaw surgery with counterclockwise rotation of the maxillomandibular complex. The patients were divided into 2 groups: group 1, 30 patients (8 men, 22 women) who underwent maxillary and mandibular advancement and group 2, 20 patients (14 men, 6 women) who underwent maxillary advancement and mandibular setback. Presurgery and postsurgery lateral cephalometric radiographs were analyzed to correlate changes in pharyngeal airway space dimensions and velopharyngeal anatomy with maxillary and mandibular positional changes. The calibration showed a more than 0.94 correlation for both intra- and interoperator error. The average follow-up time was 29.6 months in group 1 and 22.2 months in group 2. Mean maxillary surgical change at point A was 4.15 mm in group 1 and 2.5 mm in group 2. Mean mandibular surgical change at the genial tubercles was 7.5 mm in group 1 and -4.95 mm in group 2. After surgery, group 1 patients had an increase in pharyngeal airway space of 47% at the soft palate and 76% at the base of the tongue relative to the amount of mandibular advancement. Group 2 patients had a decrease in pharyngeal airway space of 47% at the soft palate and 65% at the base of the tongue relative to the amount of mandibular setback. Double-jaw surgery with counterclockwise rotation of the maxillomandibular complex significantly affects the pharyngeal airway space and velopharyngeal anatomy in patients with high occlusal plane facial morphology, with both mandibular advancement and setback. PMID- 11500658 TI - Changes in mandibular position and upper airway dimension by wearing cervical headgear during sleep. AB - We previously reported that the wearing of cervical headgear induced forward displacement of the mandible in awake subjects. However, it was unclear whether such mandibular displacement also occurred during sleep. The purpose of this study was to examine changes in mandibular position and oropharyngeal structures that were induced by the wearing of cervical headgear during sleep. Ten healthy adults (7 male and 3 female) who gave their informed consent were included in this study. A pair of lateral cephalograms was taken with the patient in the supine position with and without cervical headgear at end-expiration during stage 1 to 2 non-rapid-eye-movement sleep. The Wilcoxon signed-rank test was used for a statistical analysis. The amount of jaw opening was significantly decreased by the wearing of the cervical headgear (P <.05), although no significant anteroposterior mandibular displacement was induced. The sagittal dimension of the upper airway was significantly reduced (P <.05); however, no significant changes were observed in the vertical length of the upper airway. Although the hyoid bone and the third cervical vertebra moved significantly forward by the wearing of the cervical headgear (P <.05), the relationship among the mandibular symphysis, the hyoid bone, and the third cervical vertebra did not change. These results suggest that cervical headgear significantly reduced the sagittal dimension of the upper airway during sleep, although there was no significant anteroposterior displacement of the mandible. PMID- 11500659 TI - Cephalometric and polysomnographic analyses of functional magnetic system therapy in patients with obstructive sleep apnea. AB - The therapeutic effect of a functional magnetic system on obstructive sleep apnea and the system's operating mechanism have not been examined. Two hypotheses are postulated: a functional magnetic system increases the size of the oral cavity airway passage, or it increases the pharyngeal space. Twenty-eight patients with mild-to-moderate obstructive sleep apnea were examined; 10 patients (9 men and 1 woman; aged, 50.5 +/- 2.6 years) met the study criteria. After baseline nocturnal polysomnography and daytime tiredness self-evaluation, a functional magnetic system was inserted. The functional magnetic system is a mandibular repositioning appliance that uses a pair of attractive magnets (Sm2Co17), placed opposite each other in the jaws, which results in an advancement-to-opening ratio of 1:2. After 8 weeks of functional magnetic system treatment, polysomnography, daytime tiredness, and nighttime snoring were evaluated, and cephalogram radiographs with and without the appliance were taken. It was found that the respiratory disturbance index decreased significantly; minimal oxygen saturation increased significantly, reaching a normal value; day time tiredness improved; snoring declined; the oral cavity anterior region increased significantly, and the pharyngeal airway passages did not change. Reduction in the respiratory disturbance index and enlargement of the anterior oral cavity area were highly and significantly correlated. In conclusion, the functional magnetic system is a reliable mandibular repositioning appliance that has no apparent adverse effects. A functional magnetic system operates by increasing the anterior region of the oral cavity, mainly vertically, with no change in the posterior oral cavity region and pharyngeal airway passages. PMID- 11500660 TI - Posttreatment evaluation of the distal jet appliance. AB - This study analyzed molar distalization with the distal jet appliance, its effect on the anchor teeth, and the outcome at the completion of orthodontic treatment. Pretreatment, after distalization, and posttreatment lateral cephalometric radiographs were evaluated for 21 adolescent girls and 12 adolescent boys. The mean age of the subjects at the time of the pretreatment radiograph was 12.8 +/- 2.2 years. The mean time for the correction of the Class II molar relationship was 6.7 +/- 1.7 months, and the mean total treatment time was 25.7 +/- 3.9 months. The results of this study showed that the distal jet appliance distalized the maxillary molars, but there was significant loss of anchorage. The distal jet also showed less tipping of the maxillary molars and better bodily movement of molars because the force was applied closer to the center of resistance. The observations of treatment outcome indicate that the 1.8-mm mean net anterior movement of the maxillary first molar was more than offset by the 4.8-mm mesial movement of the mandibular first molar. There was no significant increase in lower face height. Consequently the effect on the facial balance was negligible. PMID- 11500661 TI - Can maxillary incisor trauma be predicted from cephalometric measurements? AB - Dental injury is a traumatic event related to many factors, including dentofacial morphology. In a previous study in which we used a logistic regression model, 5 cephalometric variables were defined as being the best predictors of traumatic injury. The purposes of this study were to test the predictive value of the logistic regression analysis results for the occurrence of dental injury in a new patient sample and to compare these results with the outcome based on a visual evaluation of cephalograms. The study population consisted of 2 groups of orthodontic patients: patients with injured incisors (n = 64) and patients with non-injured incisors (n = 90). Cephalometric skeletal, dental, and soft tissue measurements were made. In addition, 6 professional judges visually evaluated all cephalograms. Classification tables were constructed for the results of the logistic regression model and for the assignment of patients according to the visual evaluation of the cephalograms. The percentage of correctly classified patients at the.7 probability level was 62%. Surprisingly, the professional panel rendered an equal predictive value. The relatively inferior performance of both the mathematical model and the "eyeballing" may demonstrate that dentofacial morphology is only partially responsible for susceptibility to trauma. PMID- 11500662 TI - Experimental evaluation of initial tooth displacement, center of resistance, and center of rotation under the influence of an orthodontic force. AB - The purpose of this study was to determine the location of the center of resistance and the center of rotation of the maxillary central incisors under the influence of a single simple force and to investigate related geometric parameters of the teeth and the surrounding periodontal tissues. By measuring the initial displacement of the central incisors with a magnetic sensing system, the location of the center of resistance and the centers of rotation associated with various forces were determined in 3 human subjects. The results show that the location of the center of resistance of the maxillary central incisor depends on the palatal bone level and is at approximately two-thirds of the palatal alveolar bone height, measured from the root apex. A greater moment-to-force ratio is needed for any controlled movement of the maxillary incisors during retraction in patients with reduced palatal alveolar bone height. This study suggests a method for estimating the location of the center of resistance of a tooth. PMID- 11500663 TI - Physical properties of root cementum: Part I. A new method for 3-dimensional evaluation. AB - Cementum is a nonuniform connective tissue that covers the roots of human teeth. Investigation of the physical properties of cementum may help in understanding or evaluating any possible connection to root resorption. A variety of engineering tests are available to investigate these properties. However, the thickness of the cementum layer varies, and this limits the applicability of these techniques in determining the physical properties of cementum. Hardness testing with Knoop and Vickers indentations overcame some of these limitations, but they prohibited the retrieval and retesting of the sample and therefore the testing was restricted to one area or section of the tooth. Another limiting factor with the existing techniques was the risk of artifacts related to the embedding material such as acrylic. A new method to investigate the physical properties of human premolar cementum was developed to obtain a 3-dimensional map of these properties with the Ultra Micro Indentation System (UMIS-2000; Commonwealth Scientific and Industrial Research Organization, Campbell, Australia). UMIS-2000 is a nano indentation instrument for investigation of the properties of the near-surface region of materials. Premolars were harvested from orthodontic patients requiring extractions and then mounted on a newly designed surveyor that allowed sample retrieval and 3-dimensional rotation. This novel method enabled the quantitative testing of root surface cementum, on all 4 root surfaces, extending from the apex to the cementoenamel junction at 60 different sites. PMID- 11500664 TI - Intrusion of overerupted molars by corticotomy and magnets. AB - Although posterior tooth intrusion in an adult patient is a difficult procedure, it can be achieved without extruding the adjacent teeth by performing a corticotomy and using magnets. In carrying out this procedure on 2 adult patients whose molars had overerupted due to the early loss of antagonists, tooth movement was rapidly achieved without discomfort or side effects. PMID- 11500665 TI - Litigation, legislation, and ethics. Expanding the scope of the doctor-patient contract. PMID- 11500668 TI - Drug eruptions in children. PMID- 11500673 TI - Tracing medical information over the Internet. AB - The Internet became with do doubt a huge and valuable source of information for researchers. The wealth of information on the Internet is second to none and medical information is no exception. Yet with the vast expansion of the Internet and the World Wide Web in specie, to find the kind of information one is looking for, he/she needs to browse thousands of web sites and the experience would be like digging into a stack of hay looking for a needle. That's why search engines and subject indexes, as means to overcome this problem, were introduced and grew so rapidly. In general, there are three approaches to retrieve data from the World Wide Web; the subject directories, search engines and detailed subject indexes. However, there is no single comprehensive search engine or directory and it is recommended to use more than one with different keywords and synonymous. PMID- 11500674 TI - Update on diagnosis of congenital infection. AB - Congenial infection is one of the greatest diagnostic challenges facing clinicians. The list pathogens related to intrauterine infections continues to grow with the identification of new etiologies and resurgence of others. Identification of a congenital infection as early as possible has both diagnostic and therapeutic advantages. This article will give an overview on common clinical findings in infants with congenital infection and a recommended clinical investigational approach for suspected congenital infection PMID- 11500675 TI - New trends in prenatal screening for chromosomal abnormalities. AB - In the recent years, the scope of prenatal genetic diagnosis has expanded greatly with the development of a number of new methods, such as maternal serum screening and ultrasound screening for detection of fetal abnormalities. These methods have the advantage of providing earlier diagnosis in addition to being non-invasive and of less psychological traumas. Pre-implantation genetic diagnosis is another new growing field offering the genetic diagnosis prior to implantation. The most promising of all is the first trimester biochemical screening in conjunction with ultrasound nuchal translucency screening. PMID- 11500676 TI - Subclinical diabetic neuropathy: a common complication in Saudi diabetics. AB - OBJECTIVE: To determine the prevalence of sub clinical diabetic neuropathy in Saudi diabetics and the risk factors associated with symptomatic diabetic neuropathy. METHODS: A prospective study of Saudi diabetics attending King Abdulaziz University Hospital out patient clinic from January 1998 until April 1999. Detailed information of each patients' age, sex, body mass index, type and duration of diabetes mellitus, mode of treatment, degree of blood glucose control, presence of hypertension, hyperlipidemia, smoking, family history of diabetes mellitus and hypertension were recorded. Patients were assessed for diabetic neuropathy using the Michigan Neuropathy Program. Patients who were asymptomatic and scored less than 2 on simple clinical examination were referred to a neurologist for a complete neurological examination and nerve conduction studies. RESULTS: A total of 237 patients were studied with a mean age of 54.19 years and mean duration of diabetes 10.6 years. Symptomatic diabetic neuropathy was present in 132 (56%) patients while subclinical neuropathy was present in 58 (57%) of asymptomatic patients. Old age, type II diabetes with long duration, poor control and smoking were risk factors associated with symptomatic diabetic neuropathy (p<0.001, p=0.09, p<0.001, p=0.04, p=0.08). CONCLUSION: Subclinical diabetic neuropathy is common. Early diagnosis is important for possible prevention of late neuropathic complications (foot ulcers and infections). Prolonged poorly controlled diabetes mellitus, old age and smoking are risk factors for symptomatic diabetic neuropathy. Meticulous blood glucose control is important for nerve function protection. Researches are urgently needed for satisfactory therapy. PMID- 11500677 TI - Prevalence of diabetes mellitus in a Saudi community. AB - OBJECTIVE: The aim of this retrospective cross sectional study is to find out the prevalence of Diabetes Mellitus according to age and sex in the population attending the family and community medicine clinic in Al-Kharj Armed Forces Hospital. METHODS: Total eligible patients of Al-Kharj Military Hospital are about 100,000. The case notes in Medical Records were selected randomly and examined. Data was collected from 3747 case notes. They were divided into different age groups and also separated into male and female groups. RESULTS: The prevalence of Diabetes Mellitus in this study sample of 3747 (1,683 males and 2,064 females) was 2.55% in males and 5.32% in females. In the age group of 35 and above the prevalence in males was 8.52% and in females 19.48%. CONCLUSION: A high prevalence of Diabetes Mellitus was noted in the population and particularly in females. There is need for comprehensive programs for the prevention of Diabetes Mellitus particularly in the adult population. PMID- 11500678 TI - Diabetic foot: presentation and treatment. AB - OBJECTIVE: The aim of this study is to determine the mode of presentation, medical and surgical treatment in diabetic foot. METHODS: Medical notes of 34 diabetic patients admitted to King Abdulaziz University Hospital in Jeddah from June 1997 to June 1999 with diabetic foot were studied retrospectively. Collection of demographic data, mode of presentation, investigations, medical treatment (antibiotics cover) and surgical treatment has been studied. Data of those patients were collected for gender, age, duration of diabetes, mode of presentation, presence of peripheral neuropathy, peripheral vascular disease and other complications, precipitating factors, concurrent medical illness (ischemic heart disease, hypertension, chronic renal failure, and retinopathy), microbial flora, medical treatment by antibiotics, surgical treatment, and duration of hospital stay RESULTS: Majority of patients were male above 50 years, mean age 59 +?- 9.6, 29 male, 47% Saudi, 65% diabetic patients, their blood sugar were controlled by oral hypoglycemic drugs. However only 29% were on insulin. History of trauma preceding infection was present in 20%. Peripheral neuropathy was the main precipitating factor in 94%, as well the presence of peripheral vascular disease in 50%. Smoking was found in 44% patients. Foot ulcer was the most common type of presentation in 59% of patients, 65% of these patients need debridement. However 8 patients had major amputation. Proteus and pseudomonas were the most common organisms isolated. The mean hospital stay was 21.44 +?- 17.7 days. CONCLUSION: Diabetic foot sepsis is a common health problem presented in Saudi Arabia particularly among men, peripheral neuropathy, as well poor glycemic control are the most common precipitating factors. Foot infection is usually poly microbial. Most of these patients required debridement, however 8 (23.5%) of them, ended up having major limb amputation. PMID- 11500679 TI - Consumers' satisfaction with primary health services in the city of Jeddah, Saudi Arabia. AB - OBJECTIVE: To study consumers' satisfaction and socio-demographic correlates of satisfaction with services provided by Primary Health Care Centres in the city of Jeddah. METHODS: Study subjects included the consumers visiting the Primary Health Care Centres during the study period. Four Primary Health Care Centres were randomly selected from Jeddah according to the geographical location. Seventy five subjects were selected systematically where every tenth Saudi aged 15 years or above was chosen. Data was collected via a self administered pilot tested questionnaire which included socio-demographic characteristics, as well as the overall and differential satisfaction with the different services and facilities in the selected Primary Health Care Centres rated on a scale of 1 - 5 points, the higher the score the higher the satisfaction. RESULTS: Male subjects constituted 60% of the visitors. More than half of the subjects were young 15-29 years of age, about 58% were married, 50% completed intermediate/secondary school, more than two fifths of the subjects were employees and more than three quarters have a monthly income of 6000 Saudi Riyals or less. The summary satisfaction score was 3.76 points and the overall satisfaction with the services provided was 2.45 points out of a maximum of 5 points. The highest satisfaction was for dental clinic (3.44 points) and the lowest for co- operation of the receptionist (1.95 point). Unskilled laborers showed the highest summary and overall satisfaction (4.31 and 2.71 points) and students showed the lowest satisfaction scores (3.54 and 1.89 points). Other socio-demographic variables were not significantly related to summary and overall satisfaction scores. CONCLUSION: Measuring satisfaction by asking one summary question tends to give a significantly higher satisfaction score compared to satisfaction score taking in consideration of all services offered. Certain service components need corrective intervention measures to make them more satisfactory to consumers. PMID- 11500680 TI - Children's risk perception and parents' views on levels of risk that children attach to outdoor activities. AB - OBJECTIVE: Higher rates of outdoor injuries are found in older children, boys and children from more deprived areas. Some of the causes of these variations have been studied in some age groups and in different situations. To study children's perception of safety and danger and parents' views on the level of risk that their children attach to outdoor activities comparing age groups, boys and girls, and the less economically well off with the better off in order to understand outdoor accidental injuries. METHODS: A cross-sectional study, obtained information from children aged 7 and 9 (n=471) and their parents (n=416) were living in Newcastle upon Tyne by self-completed questionnaires. Nine schools in different socio-economic areas were selected. Children were asked to paste different stickers on to the pictures in the classrooms. The parents' questionnaires were delivered by children and then returned to school after completion. The deprivation of an individual was accessed by Z-score of the households. RESULTS: Older children compared to younger ones, boys versus girls and more economically deprived children compared to less deprived children had less perception of the dangers of outdoor activities. For example, older children compared to younger ones were more likely to perceive crossing a busy road with their friends (38.0% vs. 26.4%, P=0.007) as being safe. Boys compared to girls were more likely to perceive climbing wall (31.5% vs. 13.2%, P<0.001) as being safe. More economically deprived children were more likely to perceive cycling without a helmet (25.8% vs. 11.2%, P<0.001) as being safe than less economically deprived children. CONCLUSION: These findings may partly explain the higher rate of accidental injuries among older children, boys and more deprived children. The results may inform how education of primary schoolchildren about dealing with activities such as crossing a busy road, climbing walls and playing in street and how parents should be conscious of their children's outdoor activities. PMID- 11500681 TI - Erythrocyte sedimentation rate during steady state, painful crisis and infection in children with sickle cell disease. AB - OBJECTIVE: To evaluate Erythrocyte Sedimentation Rate in children with sickle cell disease. METHODS: Erythrocyte Sedimentation Rate was performed in 95 children with sickle cell disease during the period from December 1992 to June 1999. Patients were classified into 4 groups. Patients in the steady state (group 1), painful crisis (group 2), mild infection (group 3) and moderate/severe infection (group 4). Comparison between the groups was performed using ANOVA and ANCOVA. RESULTS: There were 55 males (58%) and 40 females (42%) with a mean age of 79.1 months. The mean Erythrocyte Sedimentation Rate values for the various groups were as follows: Group 1 (n=10); mean Erythrocyte Sedimentation Rate is 5 mm/hr (+/-4.6) and was significantly the lowest (p<0.05), group 2 (n=44); mean Erythrocyte Sedimentation Rate is 25.8 mm/hr (+/-16.3), group 3 (n=18); mean Erythrocyte Sedimentation Rate is 28.1 mm/hr (+/-25.8), while group 4 (n=23); mean Erythrocyte Sedimentation Rate is 99.2 mm/hr (+/-33.5) and was significantly the highest (p<0.05). Two cut-off values for the Erythrocyte Sedimentation Rate (> or = 30 mm/hr then > or = 50 mm/hr) were considered to evaluate their specificity and sensitivity for moderate/severe infection. CONCLUSION: Although the Erythrocyte Sedimentation Rate is low in sickle cell disease, it does increase during painful crisis and infection and it is a useful indicator of moderate/severe infection. PMID- 11500682 TI - Reinfestation in cutaneous leishmaniasis: a new look at predisposing conditions. AB - OBJECTIVE: A prospective study of cases of reinfestation with cutaneous leishmaniasis is carried out to try to identify if there is any associated condition which might predispose to reinfestation. METHODS: Over a 2 year period, cases of reinfestation with cutaneous leishmaniasis presenting in the Department of Dermatology, were collected. RESULTS: Thirteen cases showing a typical scar with a parasitologically proven new infection were included. The mean age of patients was 54.69+/-2.65 years. The time lapse between the two infections was 52.38+/-2.60 years. The scar of the primary infection was mainly on the face while the lesion was mostly on the limbs. Six patients had diabetes mellitus, one patient was pregnant and three received either oral corticosteroids or other immunosuppressive drug treatment. CONCLUSION: It is concluded that in patients with reinfestation it is important to take a careful medical history and send for a fasting blood glucose, as there is a chance of one in two that the patient may be diabetic. PMID- 11500683 TI - Inhalation of wild barley into the airways: two different outcomes. AB - OBJECTIVE: There are only two previous reports of wild barley inhalation into the tracheobronchial tree in the literature. Our aim is to describe the effects, outcome and management of wild barely inhalation into the airways. METHODS: In a retrospective study, 18 patients with inhalation of spike of wild barley into the tracheobronchial tree were divided into 2 groups according to their clinical presentation. RESULTS: Eighteen children below 5 years of age presented between 1989 and 1994 inclusive. Fourteen patients presented with a short duration of choking and cough. The wild barley spike was removed by laryngoscopy (12 patients) or rigid bronchoscopy (2 patients). Four patients presented with a longer history of cough, dyspnea and fever and had serious respiratory disease such as pneumothorax, lobar pneumonia and pleural empyema requiring surgical intervention. All patients made a satisfactory recovery. CONCLUSION: Wild barley is a common grass in our area and we should be aware that children are exposed to the risk of inhalation into the airways. PMID- 11500684 TI - Nasal bone fracture. AB - OBJECTIVE: To study fracture nasal bone at King Faisal Hospital University in Al Khobar from various aspects, to present our experience and to compare the result with previous works. METHODS: A prospective study conducted from January 1997 to May 1998 including all new cases of fracture nasal bone presented at our emergency department. Plain x-ray of the nose is requested for all patients except one in whom pregnancy is suspected. Reduction of fracture nose was carried out 2-7 days post injury by closed technique under general or local anesthesia with application of nasal cast for 2 weeks. Follow-up of those patients in our clinic is evaluated regarding shape, patency and satisfaction. RESULTS: Eighty six patients included in our study with peak incidence at age 1-30 years with male being dominant. Fall down and road traffic accidents were the common cause of fracture nasal bone. The common presentation was swelling and tenderness in all patients followed by deformity (30.5%) nasal obstruction (40%), and deviated nasal septum (15%). Negative x-ray finding was noticed in 30%, and computerized tomography scan was required for evaluating complicated cases. Only 30 patients (35%) required reduction by closed technique, 23% had poor outcome and were considered as failure. CONCLUSION: This paper stresses the importance of early evaluation of fracture nose in order to avoid complications. The diagnosis is made mainly based on clinical judgement. Closed reduction technique under general anesthesia within 2-10 days post injury is the standard treatment if needed. Failure by closed technique can be managed by septorhinoplasty months later. PMID- 11500685 TI - Khat induced hemorrhoidal disease in Yemen. AB - OBJECTIVE: The aim of this study was to evaluate the potential association between the habit of khat chewing and the development of hemorrhoidal disease METHODS: Four hundred and seventy four individuals (373 men and 101 women) with ages ranging from 17 to 80 years were divided into 2 groups. Group 1 (n=247) chronic khat chewers. Group 2 (n=200) non-khat chewers. Data was collected regarding chewing habits, colorectal symptoms, abdominal, proctoscopic, and operative findings. RESULTS: The key difference between the 2 groups was the incidence of hemorrhoids and hemorrhoidectomy. In the chronic khat chewers group: 169 (62%) had hemorrhoids. Of these 124 (45.4%) underwent hemorrhoidectomy. In the control group there is 8 (4%) had hemorrhoids and one patient underwent hemorrhoidectomy (0.5%). P-value (0.05). CONCLUSION: The study demonstrated a significant association between the habit of khat chewing and the development of hemorrhoidal disease. PMID- 11500686 TI - Undiagnosed term breech: management and outcome. AB - OBJECTIVE: To assess the antenatal and intra-partum risk factors associated with undiagnosed breech at term and to compare their outcome with those diagnosed before the onset of labor, as well as identifying the determinants for cesarean section in breech presentation. METHODS: Case control study of 183 undiagnosed breech deliveries at term with those diagnosed before labor. RESULTS: There was no significant difference between diagnosed term breech and undiagnosed breech deliveries with regards to maternal, neonatal outcome and incidence of cesarean section. Footling breech presentation, hypertension and failure to attend antenatal visits were independent risk factor for cesarean section in all case of breech. CONCLUSION: Undiagnosed breech at term are not at increased risk for cesarean section and there is no additional maternal or fetal morbidity. PMID- 11500687 TI - Munchausen syndrome by proxy. AB - Five children (3F:2M), in the age group 1 years to 11 years, with Munchausen syndrome by proxy are reported from the Sultanate of Oman. They were seen over a four years period from 1996-1999. In all these children, the mother came up with history of uncontrolled epilepsy. Carbamazepine was the most common antiepileptic drug used. One of these children remained hospitalized elsewhere for nearly 9 months, as a case of uncontrolled status epilepticus. It took 18 months to 6 years (mean 2.8 years) to establish the diagnosis and the mother was the offender in all. The main lead to diagnosis, was the disparity between history and clinical presentation to hospital. The carbamazepine levels were several times above the upper limit of therapeutic range. Munchausen syndrome by proxy very much exists here, but is possibly less recognized and needs immediate attention to formulate policies to identify and manage these children. It is necessary to create awareness even in the medical community, to recognise this problem. There is an urgent need to develop a child protection council at the national or regional level. PMID- 11500688 TI - Delayed presentation of congenital diaphragmatic hernia in association with group B streptococcus infection in a preterm Omani neonate. AB - We present an interesting case of a preterm Omani newborn that had delayed onset of congenital diaphragmatic hernia in association with group B streptococcus infection. The association and the pathogenesis are supported by literature review. The message to follow is that any neonate with prolonged course of streptococcal pneumonia, with unusual course, should be investigated for presence of congenital diaphragmatic hernia. PMID- 11500689 TI - Modified retrograde intubation in a patient with difficult airway. AB - We report a modified technique of retrograde endotracheal intubation in a patient with limited motility at the atlanto-occipital joint, temporomandibular joint, and cervical spine, presenting for closure of a large oronasal fistula. Despite more recent advances in intubation techniques and technology, retrograde intubation still deserves a place in the anesthetist's armamentarium for the management of the difficult airway. PMID- 11500690 TI - Double dislocation of the interphalangeal joints in the finger. AB - A case of simultaneous dislocation of the proximal and distal interphalangeal joints of the same digit is described. The case presented at Princess Basma Teaching Hospital after athletic trauma. It was treated successfully with close reduction followed by two weeks immobilization in slight flexion position. The condition is described in this report with review of the relevant literature. PMID- 11500691 TI - Laparoscopic juxtarenal varicocelectomy. PMID- 11500692 TI - Pseudohypoaldosteronism type 1 in an Omani male infant simulating salt-losing congenital adrenal hyperplasia. PMID- 11500693 TI - Neonatal salmonella meningitis. PMID- 11500694 TI - Overview of chronic diseases in the Kingdom of Saudi Arabia. PMID- 11500695 TI - Temporal aberrations in dreams. PMID- 11500696 TI - Criteria for diagnosis of growth hormone deficiency. PMID- 11500697 TI - Prevention of migraine. PMID- 11500698 TI - Occupational hearing loss. AB - Occupational hearing loss is a common work related problem that can be attributed to an offending agent in the workplace. This paper describes the different causes of occupational hearing loss and its compensation. Physicians should be aware of this preventable medical condition. PMID- 11500699 TI - Job satisfaction and organizational commitment for nurses. AB - OBJECTIVE: The purpose of this study is to find out the extent to which nurses in public hospitals are satisfied with their jobs and committed to their hospitals. Also, it is intended to assess the relationship between job satisfaction and organizational commitment, as well as to examine the effect of nurses' demographic factors on these 2 variables. METHODS: Two-hundred and ninety nurses working in a number of public hospitals are the sample size. The measures of the study are already tested for validity and reliability. In this study, they are also examined and found to be valid and reliable. The method of data collection used was the questionnaire to include a large sample. RESULTS: The means and standard deviations show that nurses are satisfied with their jobs to some extent, and they are slightly committed to their hospitals. A strong positive correlation between job satisfaction and organizational commitment was found at 0.59 (p <.01). Also, age is significantly correlated with satisfaction and commitment, but experience is correlated only with the latter. Moreover, analysis of variance shows that nurses differ in their degree of commitment in terms of their marital status and nationality, whereas they differ in their degree of satisfaction only with regard to their nationality. CONCLUSION: This study found that nurses in public hospitals are slightly satisfied and committed to their hospitals. Besides, satisfied nurses tend to have a higher degree of commitment than less satisfied ones. However, further research is needed to examine this result before a generalization is made. PMID- 11500700 TI - Growth hormone: do we have a national perspective of indications for its use? AB - OBJECTIVE: Presentation of indications for growth hormone use in children, from the view of physicians practising in Saudi Arabia. METHODS: A questionnaire containing a list of common possible indications of growth hormone use was collected from 52 physicians practising in Saudi Arabia who were attending a didactic endocrinology course. RESULTS: All (100%) physicians considered growth hormone deficiency to be an indication. Sixty four and 29% considered Turner's syndrome and chronic renal failure to be indications. Other indications included Russell-Silver syndrome in 23%, X-linked hypophosphatemic rickets in 10%, Achondroplasts in 10%, Sickle cell anemia in 10% and Bartter's syndrome in 2%. Genetic and constitutional short stature were considered in 17 and 19%. Only 65% did not advocate the use of growth hormone for short non-growth hormone-deficient children. CONCLUSION: Indications of growth therapy are not clearly defined in Saudi Arabia. It is indicated to define this by a clear, national decided criteria which should take into consideration the internationally approved indications, availability and cost of this hormone. PMID- 11500701 TI - Common causes of admission in diabetics. AB - OBJECTIVE: To report on the causes of admissions of diabetic patients to the medical unit of King Abdulaziz University Hospital, mortality and risk factors, associated with high mortality, and to find out possible ways to reduce admissions, cost and mortality. METHODS: Retrospective analysis of diabetic admissions to the medical unit of King Abdulaziz University Hospital between January 1996 to September 1999. Patients age, sex, body mass index, type and duration of diabetes mellitus, mode of treatment, degree of blood glucose control, presence of hypertension, hyperlipidemia, and smoking were recorded as well as the causes of admissions and mortality. RESULTS: A total of 5917 patients were admitted, 17% of them were diabetics. Admissions for blood glucose control and for macrovascular complications were found in 21% and 38%. Mortality rate was 13%. Hypertension, hyperlipidemia, smoking, obesity, infection, poor glycemic control, long duration of diabetes mellitus and long hospital stay were risk factors associated with high mortality. CONCLUSION: Macrovascular complications and uncontrolled blood glucose were the most common causes of admissions. Control of hypertension, hyperlipidemia, cessation of smoking and weight reduction will not only decrease the risk of macrovascular complications, but also in addition to patient's education for tight blood glucose control, will decrease the rate, cost and mortality of diabetic admissions. PMID- 11500702 TI - Effect of labor duration on fetal fibrinolysis: a pilot study. AB - OBJECTIVE: Activation of neonatal and maternal fibrinolysis at birth has been shown earlier. To evaluate further on the effect of labor duration on the fetal fibrinolysis, we conducted this preliminary study. METHODS: An indirect assessment of activation of fetal fibrinolysis was carried out, by measuring the cord blood D-dimer. The cord blood samples were obtained randomly from 11 women undergoing vaginal delivery at term. Specimens were analyzed within 2 hours by using latex agglutination test. Pregnancy details were obtained from the delivery ward register. RESULTS: The mean duration of labor was noted to be 5.4 hours (3.5 9.46). The mean maternal age was 25.5 years (21-34). The mean gravidity was 2.4 (1-5) and the mean parity was 1.4 (1-4). No effect of duration of labor on the cord blood D-dimer concentration was observed. CONCLUSION: Our findings suggested that it is not the time or duration of labor process that is involved in the activation of fetal fibrinolysis. The role of uterotropins and uterotonins (oxytocin, prostaglandins and endothelin-1), in activation of fetal fibrinolysis, should be evaluated in further studies. PMID- 11500703 TI - Clinical characteristic of amoebic liver abscesses in the North of Iraq. AB - OBJECTIVE: The purpose of the study was to find out the clinical characteristic of amoebic liver abscesses in this area, the simplest method for diagnosis and to determine the effectiveness of treatment by metronidazole therapy proved by disappearance of symptoms and regression in the size of the abscesses by ultrasound. METHODS: We studied prospectively all cases of suspected liver abscesses admitted to our unit over 2 years (1990 & 1991). A special case sheet was prepared. Daily follow up of patients was carried out in hospital for at least 10 days. Patients were later followed up by ultrasound after discharge. We compared the rate of infection from the hospital records over the last 9 years up to the end of 1998. RESULTS: We found the clinical features of the disease similar to those mentioned in essential text books of medicine, except that pain is not always epigastric while fever may be absent and hepatomegaly is not marked. Ultrasound is a simple, cheap diagnostic test, which is available. Metronidazole is an effective treatment and none of the patients required an invasive method for diagnosis or treatment. None had secondary bacterial infection. CONCLUSION: Amoebic liver abscess if untreated is a grave disorder. We concluded that recognizing the disorder clinically and proving it by ultrasound is the main method for diagnostic confirmation. Following the hospital records of the last 9 years, it seems that the rate of infection is declining in this area. PMID- 11500704 TI - Neonatal meningitis. AB - OBJECTIVE: To determine the prevalent bacterial agents of neonatal meningitis and their antibiotic susceptibility in a referral intensive care unit in Assir Central Hospital, Saudi Arabia, during the years 1993-1998. METHODS: Records of newborn infants with positive cerebrospinal fluid culture during the period were retrospectively studied. RESULTS: There were 1473 nursery admissions, of which 32 episodes of meningitis occurred amongst 31 neonates. Klebsiella pneumoniae (31%) and Serratia marcescens (21%) were the main pathogens. The incidence of concurrent septicemia among these infants was 58%. Klebsiella pneumoniae appears to dominate in both early and late onset infections. The sex incidence was equal and the mortality rate was 48%. CONCLUSION: The survey identifies Klebsiella pneumoniae and Serratia sp. as the leading bacterial agents of neonatal meningitis in our environment. The relatively high frequency of Serratia infection in the present survey appears unique as this organism is comparatively rare in other reports across the globe. No Group B Streptococcus was isolated, which is in contrast to reports obtained in Europe, America and Australia where it is the predominant organism of neonatal sepsis or meningitis. Antibiogram identified imipenem and cefotaxime as the empirical antibiotics in infants with a clinical diagnosis of neonatal sepsis in our hospital; no more conventional use of ampicillin. In view of the changing bacterial pattern of infant infection with time even in the same environment, a periodic review of this subject is advocated. PMID- 11500705 TI - Epidemiological patterns of scoliosis in a spinal center in Saudi Arabia. AB - OBJECTIVE: To study the patterns of scoliosis seen in a major Spinal Unit in Saudi Arabia. METHODS: Medical records of 192 Saudi patients with scoliosis seen between 1992 and 1997 at the Spinal Unit of the Riyadh Medical Complex (which is a major spinal unit in the Kingdom) were studied and analyzed and epidemiological data was collected. RESULTS: Fifty nine percent of all cases of scoliosis were idiopathic, 7% were secondary to poliomyelitis and 17% were congenital scoliosis. The mean age of discovery of idiopathic scoliosis was 12.5 years and at presentation was 16 years. The mean Cobb's angle at presentation was 58 degrees C. Adolescent type constituted 74% of idiopathic curves with a male to female ratio of 1:3.8. Thoracic curves were the most common followed by the double major curves. Half of the curves were right sided. Infantile idiopathic scoliosis constituted 8% with male to female ratio of 4:1 and here left thoracic curves formed 50%. The juvenile type constituted the remaining 18% with male to female ratio of 1:1.5. CONCLUSION: The pattern of scoliosis in our study is comparable in many aspects with other studies carried out elsewhere, with the exception of delay in patients age at referral. We believe that our data could reflect a possible pattern of the epidemiology of scoliosis in Saudi Arabia, given the fact that there is no National Registry. To avoid delay in management, community education will help in the early detection of cases. PMID- 11500706 TI - Prevalence of hypertension in patients attending Al-Kharj Military Hospital. AB - OBJECTIVE: This study was carried out to ascertain the prevalence of hypertension in our community with a view to forming a baseline and planning strategies for prevention and control. METHODS: A cross sectional study carried out by collecting data from randomly selected case notes of 3747 patients (1683 males and 2064 females), regarding age, sex, blood pressure measurements and whether or not they were on treatment for hypertension. RESULTS: The prevalence of hypertension in the total population was 3.1% (females 3.05% and males 2.67%). With regard to age group, the prevalence was highest in 65-74 years in both males and females. CONCLUSION: There is a need for health education among our patients regarding the nature of hypertension and the relation between life style and hypertensive disease. PMID- 11500707 TI - Peak expiratory flow rate in Saudi school boys at Al-Khobar City, Saudi Arabia. AB - OBJECTIVE: To determine the normal peak expiratory flow rate for Saudi school boys living at Al-Khobar City. METHODS: A self administered questionnaire was completed by the parents of 1312 school boys who satisfied the selection criteria of the study. Peak expiratory flow rate was measured for them, using Wright peak flow meter. The multiple linear regression equation for peak expiratory flow rate was generated where height, age, weight, socio-economic class, presence of smoker, pets or both at home were the independent variables RESULTS: Height, age and weight were found to correlate significantly with peak expiratory flow rate. However, only height and age were included in the final regression model which had a reasonably high coefficient of multiple determination value (R2 =0.72). CONCLUSION: Peak expiratory flow rate values in this study were lower than those reported from Riyadh, other Arab countries, Europe and North America. However, they are close to those reported from Yanbu (Saudi Arabia). Normal values of peak expiratory flow rate for Saudi children should be developed. PMID- 11500708 TI - Head and neck squamous cell carcinoma in Hajjah, Yemen. AB - OBJECTIVE: Incidence of head and neck squamous cell carcinoma seems to be relatively high in Yemen but not well documented. The purpose of this study is to analyze the clinical profile of the Yemeni patients of squamous cell carcinoma of the head and neck and to evaluate the possible relationship to kath chewing. METHODS: With the help of a special protocol, all the patients of head and neck squamous cell carcinoma seen between October 1997 and December 1998 at the Ear, Nose and Throat and Dermatology Clinics of Saudi Hospital, Hajjah, Yemen Republic were subjected to detailed analysis. The diagnosis was confirmed by histopathologic studies in all the cases. RESULTS: All the 36 patients (23 male and 13 female) were Yemani nationals, aged 18 to 80 years (median age 50 years). Thirty patients were Kath addicts. The tumor was localized to the oral cavity in 17 (47%) patients, oropharynx in 1 (3%) patient, nasopharynx in 15 (42%) patients and larynx in 3 (8%) patients. CONCLUSION: The incidence of head and neck squamous cell carcinoma seems to be relatively high, especially the oral squamous cell carcinoma, all of whom had a habit of kath chewing, which may be considered as an important contributing factor. PMID- 11500709 TI - Ultrastructural study of the gastric mucosa and Helicobacter pylori in duodenal ulcer patients. AB - OBJECTIVE: To investigate the relationship between Helicobacter pylori and gastric mucosa in control and duodenal ulcer patients at the electron microscopic level. METHODS: Three antral biopsies were taken from each of 20 normal control volunteers and 30 duodenal ulcer patients presented to the gastroenterology unit at Jordan University Hospital for upper endoscopic examination. Each specimen was fixed and processed for electron microscopic study. RESULTS: Two types of Helicobacter pylori were observed and identified by their morphology at electron microscopy. The first one was characterized by double external smooth membranes and homogeneous cytoplasmic contents, and the second type with a characteristic ring-shaped intracytoplasmic vacuole. Electron microscopic examination of normal controls showed normal gastric mucosa and a small number of Helicobacter pylori in 12 out of 20 controls. However, in duodenal ulcer patients, 5 different patterns of interaction between the Helicobacter pylori and gastric mucosa were observed in relation to the severity of the disease. In duodenal ulcer patients, various types of epithelial damage was seen accompanied with a decrease or absence of mucous secretion and with more colonization of bacteria. CONCLUSION: The morphology and pathogenesis of Helicobacter pylori was described in duodenal ulcer patients, and 5 different patterns of contact between Helicobacter pylori and surface epithelium were recognized causing variable degrees of microvillous atrophy and reduced mucous secretion. The vacuolated type of Helicobacter pylori was more adherent to the damaged epithelium and there was a direct relationship between the epithelial damage and bacterial load. In the normal controls, no epithelial damage and scanty bacteria were observed. The various types of epithelial changes of gastric mucosa has initiated more research at electron microscopic level on the immune mechanism of the gastric mucosa to determine the underlying cause of the varying severity of the disease. PMID- 11500710 TI - Development of contralateral pleural effusion during chemotherapy for tuberculous pleurisy. AB - Paradoxical worsening of tuberculous lesions, despite effective chemotherapy, has been reported in intracranial tuberculomas, lymph nodes, pulmonary disease, and tuberculous pleurisy. However, development of contralateral pleural effusion during treatment of tuberculous pleurisy is very rare. We report the case of a 22 year old female patient who presented with right sided pleural effusion and was treated with antituberculous drugs. Four weeks later although her right sided pleural effusion was subsiding she developed a left sided pleural effusion. Closed pleural biopsy on the left side showed granulomatous inflammation with early caseation. Antituberculous drugs were continued and a short course of oral prednisolone was added. She recovered completely and her chest x-ray became normal after finishing her treatment. PMID- 11500711 TI - Neurobrucellosis. AB - Brucellosis, which is endemic in Saudi Arabia, can present as a disease with multisystem involvement. Occasionally the organisms invade the central nervous system producing varied neurological manifestations. Often the clinical diagnosis of neurobrucellosis is not straight forward and is obscure. Certain criteria have to be fulfilled for the definitive diagnosis of neurobrucellosis. We report 2 cases of neurobrucellosis which were encountered for the first time during the past 10 years at this hospital. PMID- 11500713 TI - Trichobezoar: a rare cause of acute bowel obstruction. AB - A surgically successfully treated case of (2.8 kg) Trichobezoar presenting with severe epigastric pain, vomiting, associated with anemia and dark stool of one days duration. Complicated on admission by an acute intestinal obstruction. There has been no report of such a case from Saudi Arabia up to the present time. The presenting symptoms and signs, and management strategies are discussed. PMID- 11500712 TI - Allergic fungal rhinosinusitis: report of 4 cases from Saudi Arabia. AB - Allergic fungal rhinosinusitis is a newly recognized clinical entity of chronic rhinosinusitis. Over the past 3 years, 4 such patients were treated in our hospital. The clinical and pathological features of these 4 cases which merited the criteria for such diagnosis, are described. All the 4 cases had history of nasal polyps, asthma, or both with radiographical evidence of pansinusitis. Histologically, the thick greenish-brown inspissated material specimens which were collected and submitted to the laboratory showed, eosinophils, Charcot Leyden crystals but no fungal elements were detected on routine hematoxylin and eosin sections and no tissue invasion was noted. However, scanty aspergillus hyphae were detected on sections stained with silver. All 4 cases grew Aspergillus flavus only from the swabs and no other fungi were seen and all were treated by surgical debridement, aeration, oral itraconazole with no steroids. PMID- 11500714 TI - Fibroadenoma of the supernumerary breast of the axilla. AB - Supernumerary breast or polymastia is a well documented anomaly of the breast, and commonly presents along the embryonic milk line extending between the axilla and groin. However, cases of polymastia have been recorded in the face, vulva and perineum. The clinical significances of these anomalies include their susceptibility to inflammatory and malignant changes, and their association with other congenital anomalies of the urinary and cardiovascular systems. The present article reports a case of fibroadenoma developing in the supernumerary breast of the right axilla in a 28 year old woman. Clinical and mammography examination of both breasts revealed no abnormalities and no lymph nodes were detected in the axillae or the neck. No associated urologic or cardiovascular abnormalities were found, and the histopathological examination of the excisional biopsy samples showed a well-defined, capsulated intracanalicular type of fibroadenoma similar to that of eutopic mammary tissue. The article also outlines the common congenital anomalies of the breast, and emphasizes on their proper clinical assessment for any other associated anomaly together with adequate surgical excision and regular follow up of the treated patients. PMID- 11500715 TI - Neonatal circumcision, motivation and technique. PMID- 11500716 TI - Unforgettable consultation. PMID- 11500717 TI - Hypernatremic dehydration in a neonate due to high sodium concentration in breast milk and apparent lactation failure. PMID- 11500718 TI - Have you ever wondered? PMID- 11500719 TI - Gastric volvulus in infancy and childhood. PMID- 11500720 TI - Hypercalcemia due to parathyroid adenoma. PMID- 11500721 TI - Pre-surgical evaluation for epilepsy surgery. AB - Neuropsychological testing may reflect subtle structural changes that may not be readily apparent with neuroimaging studies, and physiologic disruption of normal neural function secondary to epileptic activity. Neuropsychological testing is used during the pre-operative evaluation for epilepsy surgery to assess functional brain status, which, in turn, provides important information on the risks for post-operative neuropsychological deficits and also provides confirmatory evidence of seizure onset laterality in patients whose seizures originate in temporal lobes. This review will focus primarily on the pre operative neuropsychological of candidates for temporal lobectomy surgery since they represent the majority of individuals undergoing ablative epilepsy surgery, and also because the literature and knowledge for the neuropsychology of temporal lobectomy far exceeds that of any other epilepsy surgical group. PMID- 11500722 TI - Febrile convulsions in children. AB - The term febrile convulsion is not a diagnostic entity. It simply describes any seizure that occurs in response to a febrile stimulus. It usually occurs between the age of 3 months and 5 years and occurs in 2-4% of young children. The typical febrile convulsion is a generalized tonic clonic seizure lasting between a few seconds and 15 minutes, followed by a period of drowsiness. Febrile seizures tend to occur in families, although the exact mode of inheritance is not known. Viruses are the most common cause of illness in children admitted to the hospital with a first febrile seizure. Routine laboratory studies are not indicated for patients who have febrile seizures and should be performed only as part of the evaluation for a source of fever. Prognosis is generally good. Only a small minority of children develop epilepsy or recurrent non-febrile seizures. Children with febrile seizures are at no greater risk of intellectual impairments than their peers. Treatment to prevent recurrence has not been shown to prevent later development of epilepsy. PMID- 11500723 TI - Clobazam for the treatment of intractable childhood epilepsy. AB - OBJECTIVE: Clobazam is a newer 1,5-benzodiazepine used for the treatment of epilepsy. It is better tolerated and less sedating than other benzodiazepines. It has yet to gain wide use for epilepsy in the Middle East. Our objective is to report our experience with clobazam for the treatment of childhood epilepsy. METHODS: A cohort of children with intractable epilepsy, defined as recurrent seizures after at least 3 anti-epileptic medication trials, were included prospectively. Clobazam was added to a maximum dose of 2 mg/kg/day. Follow-up by two pediatric neurologists was performed. Therapeutic response was recorded as complete (no seizures), good (>50% seizure reduction), fair (<50% seizure reduction), or none. RESULTS: Thirty one children (21 males-10 females), aged 2 months-15 years (mean 4.6 years) were followed for 3-12 months. Most children (68%) had daily seizures and were on multiple anti-epileptic drugs (mean 2.3, +/- SD 1). Fourteen (45%) children had Lennox-Gastaut Syndrome. After the introduction of clobazam, 11 (35.5%) became completely seizure free and 14 (45%) had >50% seizure reduction. Side effects were reported in 7 (22.5%) in the form of excessive sedation, vomiting, irritability, behavioral change, and ataxia. In 4 children these side effects resolved either spontaneously or with dose reduction. CONCLUSION: Clobazam is a well tolerated, safe, and very effective antiepileptic drug. It has a broad spectrum of antiepileptic activity, minimal side effects, and is relatively inexpensive. Wider use of this drug is recommended in children with intractable epilepsy. PMID- 11500724 TI - Morphologic patterns of male infertility in Saudi patients. A University Hospital experience. AB - OBJECTIVE: To determine the predominant histopathological patterns seen in the testicular biopsies taken during the investigation of male infertility and to compare the obtained histopathological findings with those seen in other similar studies. METHODS: This is a retrospective study performed on 230 testicular biopsies which were examined in the Department of Pathology at King Khalid University Hospital in Riyadh over a period of 10 years. The histopathological findings were grouped into 8 different morphologic categories. We have utilized a classification that is principally morphologic but that uses known or suspected clinical associations in the case of karyotypic abnormalities and excurrent duct obstruction. RESULTS: Of the total of 230 testicular biopsies studied, 72 cases showed normal spermatogenesis, of which 50 cases were suspected to be associated with excurrent duct obstruction. Germinal cell aplasia with and without focal spermatogenesis was found in about 90 cases. Thirty cases showed hypospermatogenesis, 25 cases showed maturation arrest and 12 cases showed end stage tubular sclerosis with interstitial fibrosis. Only one case was noted to show features associated with karyotypic abnormalities. CONCLUSIONS: A higher percentage of germinal cell aplasia was noted in this study when compared with other similar investigations including one previous local study. Possible causes of these discrepancies may be related to several factors including environmental effects. The design of the different studies and the criteria used for patient selection or both could also explain the cause of these observed differences. PMID- 11500725 TI - Gastrointestinal parasites presentations and histological diagnosis from endoscopic biopsies and surgical specimens. AB - OBJECTIVE: To diagnose parasitic ova or worms during endoscopy, surgery or by histology from endoscopic or surgical specimen in our patients. METHODS: The diagnosis of parasitic disease in our patients was made by histological examinations from biopsies obtained either during an upper gastrointestinal or lower gastrointestinal endoscopy or from surgical specimens. RESULTS: Parasites were seen in endoscopic biopsies from upper gastrointestinal tract in 21 patients. Schistosoma ova was seen in biopsies from stomach or duodenum (12 patients). Small intestine biopsies showed Giardiasis (8 patients) and strongyloides (1 patient). Colonic biopsies showed schistosoma ova by paraffin section or by squash technique in 216 patients. Surgical specimens from 12 patients, who presented with acute abdomen and had surgery, due to appendicitis in 8 patients, in whom specimens showed (Schistosoma in 5 patients, amoebiasis in 2 patients and Trichuria in 1 patient). Four other patients presented with acute abdomen, where ischemic bowel necrosis or mesenteric vein thrombosis was found during surgery, specimens showed schistosoma ova. CONCLUSION: The diagnosis of gastrointestinal parasites is not only made by stool but the diagnosis can be made by histology from endoscopic biopsies or surgical specimens. PMID- 11500726 TI - Schistosomes infection rate in relation to environmental factors in school children. AB - OBJECTIVE: An epidemiological comparative survey aimed at determination of prevalence and focal distribution of Schistosomes infection and intestinal parasites to provide a reference for evaluating the need for community intervention. METHODS: All children of 14th October Primary School were involved. The children were from 7 villages that lie on the Assahul valley of lbb governorates in Yemen. The total number was 230 with (82%) boys and (18%) girls. Their age was between 5-18 years with a mean of 10.24 +/- 2.6 years. Millipore and modified Kato techniques were adopted to quantify urinary and intestinal Schistosomes eggs. Other ova, larvae, cysts were recorded whenever seen. RESULTS: It was revealed that there was a Schistosomes infection rate of 37%. The mansoni prevalence was 35%, hematobia was 5% and mixed infections were 3%. Light infection was classified among 17% of all children; moderate infection among 18% and no intense mansoni infection was determined. Whereas in the case of hematobia species, 2% were intense and 3% were light. Intensity in all children was 5% eggs/g feces in case of intestinal bilharzia and 1% egg/10 ml urine in case of urinary. With regard to the prevalence of any soil-transmitted parasites, it was found to be 69% (Ascariosis 68%, Trichuriosis 10%). Double infection was found in 10%. Hookworm eggs were not seen. Infection rates with other parasites were as follows: Giardiosis 18%, Amoebiosis 14%, ova of Hymenolepes nana were seen in 13%, Taeniosis affected 13% and E. Vermicularis 1%. CONCLUSION: Bivariate analysis revealed significant associations between Schistosomes infection with residence near the valley, male sex and frequent water contact activities. No significant association was found with the age of the child, parents' education, availability of latrine or household standpipe water. In conclusion, schistosomosis was moderate, whereas soil transmitted helminthosis were intense. PMID- 11500727 TI - Serum cytokines levels in Graves' disease. AB - OBJECTIVE: The aim of the study was to look at the serum cytokines profile in newly diagnosed thyrotoxic patients with Graves' disease and to compare their cytokine levels with those of normal control subjects. Furthermore, the levels of T4, being an indicator of the severity of thyrotoxicosis, were also correlated with the Th1/Th2 and proinflammatory cytokines in Graves' disease patients. METHODS: Serum IFN-gamma (Th1), IL-10 (Th2), inflammatory cytokines including IL 6, TNF-alpha, sCD23 and sIL-2R cytokine levels were measured in 28 patients with Graves' thyrotoxicosis and in 30 normal controls. RESULTS: In Graves' disease patients, the levels of IFN-gamma (mean 142.1 +/- 29.53 units/ml), IL-10 (mean 583.8 +/- 253.3 pg/ml) and IL-4 (mean 132.4 +/- 44.52 pg/ml) were significantly higher than their corresponding levels in controls: IFN-gamma (mean 31.6 +/- 2.08 units/ml, P<0.001), IL-10 (mean 69.8 +/- 31.72 pg/ml, P<0.001) and IL-4 (mean 46.44 +/- 11.53 pg/ml). There was a marked increase in proinflammatory cytokines in Graves' disease patients: levels of IL-6 (481.5 +/- 192.3 pg/ml) and TNF-alpha (30.69 +/- 16.7 pg/ml) were significantly higher than those of normal controls for IL-6 (63.81 +/- 21.72 pg/ml, P<0.001) and TNF-alpha (8.81 +/- 1.72 pg/ml, P<0.001). Similarly the levels of sCD23 (mean 164 +/- 67.03 ng/ml) and sIL-2R (mean 2131 +/- 461.1 units/ml) were significantly higher in GD patients than in the control group (mean 31.24 +/- 11.53 ng/ml, P<0.001) and (mean 345.53 +/- 121.75 units/ml, P< 0.001) for sCD23 and sIL-2R. Furthermore, in thyrotoxic Graves' disease patients, we detected a positive correlation between free T4 and sIL-2R levels (r2 = 0.81, P<0.00), but no significant correlation was found between T4 and the other measured cytokines. CONCLUSION: The elevated serum cytokines of Graves' thyrotoxic patients reflect the activation and interplay of mixed Th1 and Th2 cells which may be consistent with long standing inflammatory and destructive processes of thyroid gland. The clinical severity of hyperthyroidism in Graves' patients only correlated with sIL-2R. PMID- 11500728 TI - D-Xylose, fat load test and antigliadin antibody in children with coeliac disease. AB - OBJECTIVE: To study the clinical pattern of coeliac disease in children living in the northwest coast of Libya, to investigate the sensitivity and specificity of D Xylose, fat load test and antigliadin antibody and their correlation to each other and changes in jejunal biopsy. METHODS: A 4 year prospective study began in 1993. It includes all suspected coeliacs referred to our department for further evaluation. All have suction jejunal biopsy using Crosby capsule performed for diagnosis, screened at the same time with one hour D-Xylose, 3-5 hours urinary D Xylose, fat load test and antigliadin antibody and followed up for 2-4 years. RESULTS: The mean age at presentation was 8 years. There was no significant correlation between D-Xylose, fat load test and antigliadin antibody. Antigliadin antibody has 87.5% sensitivity and 50% specificity in this study. Three hours urinary D-Xylose has 93% sensitivity where as one hour D-Xylose has 82% sensitivity and only 25% specificity, whereas fat load test has 69% sensitivity. CONCLUSIONS: Blood and urinary D-Xylose and fat load tests are not useful for diagnosing coeliac disease. They have no correlation to each other or to jejunal histology. Antigliadin antibody is superior to the above tests. PMID- 11500730 TI - Patients' expectations, satisfaction and future behavior in hospitals in Riyadh City. AB - OBJECTIVE: The main objectives of this study are to assess patient satisfaction at both Ministry of Health and private hospitals in Riyadh city, to compare the perceived satisfaction of patients to their expectations prior to admission at both sectors, and to determine the variables that influence the patient's future intention. METHODS: A self-administered questionnaire was used to collect data from 500 patients (392 were returned fully completed) in 7 private and Ministry of Health hospitals. In addition to the descriptive statistics, paired and independent t-tests, phi and Cramers' V tests were used for the inferential statistics of the data analysis. RESULTS: The results showed a significant difference between the general mean scores of expectation and satisfaction levels among the private sector, as well as the Ministry of Health patients. Though 12 variables were significantly less than expected among Ministry of Health patients, 6 were significantly associated with the Ministry of Health patient's future behavior. They were staff kindness, waiting time, cleanliness of the hospital, perceived nurses' quality, perceived physicians' quality, and availability of advanced medical technology. Yet, 11 variables were significantly less than expected, only 3 variables showed significant influence on the private patient's future behavior. They were availability of medicine or pharmacy, availability of advanced medical technology, and staff kindness. CONCLUSION: This study showed the important areas stand behind the dissatisfaction of patients at both sectors. In addition the study revealed the most important area that influence patient future intention for each sector. This could help hospital managers, at both sectors, in focusing their corrective effort on such areas to improve their hospitals' service's quality. PMID- 11500729 TI - The relation of low birth weight to psychosocial stress and maternal anthropometric measurements. AB - OBJECTIVES: To determine the relationship between low birth weight and psychological stress during pregnancy, as well as anthropometric measurements of Saudi mothers. METHODS: A total of 500 Saudi women were selected, according to weight and gestation age of their new born infants, one control was selected per one case (their weight less than 2500 grams). Data was collected from El-Shamasy Maternal and Child Hospital at Riyadh city. Four instruments were used; interview questionnaire, psychosocial scales to measure life stresses, true sheet to assess maternal anthropometric data and new form characteristics. Data was analyzed using SAS system. Analysis techniques included chi-square, odd ratio and P values at < 0.05 significance level. RESULTS: The results reflected an increased risk of low birth weight newborns among women who had experienced an intermediate level of stress during a period of 12 months prior to delivery. There was also an association between low birth weight newborns and maternal anthropometric measurements such as weight, height, body mass index, and upper arm circumference. CONCLUSION: Saudi women who had an increasing amount of stress during pregnancy, under weight and short stature have an increased risk of having low birth weight babies. More research is needed in different regions in the Kingdom of Saudi Arabia by using the same test for stressor during pregnancy or even using different tests for measuring life stress during pregnancy. PMID- 11500731 TI - Hemoglobin H disease in the eastern region of Saudi Arabia. AB - OBJECTIVE: Alpha-thalassemia is frequently encountered in eastern Saudi Arabia. We wanted to find out laboratory based incidence and laboratory features of Hemoglobin H disease in the Dammam region. METHODS: We retrospectively analyzed the results of Hemoglobin electrophoresis carried out during the last 5 years in our laboratory. Hemoglobin electrophoresis was performed on cellulose acetate, pH 8.6 using Helena or Biomidi kits. Hemoglobin S was confirmed by sickle solubility test. Variant hemoglobin if present, was confirmed by citrate agar (pH 6.0) electrophoresis. Helena rapid electrophoresis system was used for plate densitometry. The diagnosis of Hemoglobin H disease was made on the basis of the presence of Hemoglobin H on electrophoresis supplemented by demonstration of Hemoglobin H inclusions in red blood cells. RESULTS: Fifteen thousand, four hundred and ninety two blood samples were analyzed by Hemoglobin electrophoresis. We found 100 cases of Hemoglobin H disease, only one case was non-Saudi. The age ranged between 45 days to 85 years. There were 51 females and 49 males. Children (less than 12 years) were 35 and of adults there were 65. There were 35 adult females and 30 adult males. The mean +/- standard deviation of Hemoglobin H in children was 13.54 +/- 7, in adult females the mean +/- standard deviation of Hemoglobin H was 12 +/- 5.4, and in adult males it was 11.99 +/- 6.4. The Hemoglobin H inclusions seen in red blood cells ranged from 2.6-80 in children and 10-80 in adults. The sickle cell trait was co-existent in 7 cases. Hemoglobin Bart's along with Hemoglobin H was seen in 32 cases. Hemoglobin F was present, beyond first year of life in 34 cases. The Hemoglobin A2 as measured by densitometry was significantly low in all of the 3 age groups as compared to corresponding controls. The complete blood count results were available for analysis in only 26 cases of Hb H disease. The mean +/- SD values of Hb (g/dl), Hct (ratio), MCV (fl), MCH (pg) MCHC (g/dl), RDW-SD (fl) and RDW-CV (%) in these patients (all age groups together) were 8.15 +/- 1,.278 +/-.04, 59.4 +/- 5.8, 17.65 +/- 2.1, 29.4 +/- 1.7, 37.8 +/- 8.7 and 25.1 +/- 4.6. The mean Hb, Hct, MCV, MCH and MCHC were significantly reduced in all 3 age groups as compared to corresponding controls. RBC counts and RDW-CV were elevated in Hb H disease compared to corresponding controls. The blood film showed typical red cell morphology. CONCLUSION: Hb H disease is not infrequently encountered in the Dammam region. This condition should be kept in mind while evaluating patients for anemia. The genetic studies to determine the exact alpha-thalassemia determinants producing Hb H disease in eastern Saudi Arabia are needed. PMID- 11500732 TI - Tracheobronchial foreign bodies in children. AB - OBJECTIVE: The objective of this study is to prove that rigid bronchoscopy is a relatively safe procedure in experts' hands and should be carried out in children whenever symptoms like persistent cough, chest infection or stridor persist despite proper antibiotic therapy. METHODS: A review of 534 bronchoscopies in children was carried out between August 1988 and May 1995. Three hundred and fifteen were male and 219 were female children. The most common age was between 1 and 2 years while the mean age being 1 year 10 months. RESULTS: Out of 534 bronchoscopies, 332 were positive for foreign bodies in the tracheosbronchial tree and 202 were negative. The most common site of lodgment of foreign body was the right bronchus. The clinical presentation was variable. An increase in the number of cases was observed. A variety of foreign bodies were encountered, the majority being peanuts. CONCLUSION: In our opinion rigid bronchoscopy is a relatively safe procedure and should be carried out in children whenever symptoms like persistent cough, chest infection or stridor persist despite proper antibiotic therapy. PMID- 11500733 TI - Temporomandibular joint osteoarthrosis. Histopathological study of the effects of intra-articular injection of triamcinolone acetonide. AB - OBJECTIVES: The aim of this study was to investigate the histopathological changes of human temporomandibular joint osteoarthrosis following intra-articular injection of triamcinolone acetonide, and compare the results to these of control group. METHODS: One hundred and forty nine patients who complained of temporomandibular joint dysfunction underwent non-surgical treatment for at least 6 months. Twenty three patients had high condylectomy as a treatment. Forty four patients received two intra-articular injections of triamcinolone acetonide. Histological study was carried out on 44 specimens from 41 patients, the histological changes were recorded for each specimen. RESULTS: Patients who received temporomandibular intra-articular injection of triamcinolone acetonide showed damage to the fibrous layer (100%), to the cartilage (64%) and to the bone (42%). In those cases treated by condylectomy there were changes in the fibrous layer (100%), in the cartilaginous layer (68%) and to the bony layer (32%). Analysis of the results showed that the changes seen in the steroid (triamcinolone acetonide) group were more obvious than the changes in the non steroid group. CONCLUSION: The result of this study indicated that intra articular injection of steroid (triamcinolone acetonide) into human osteoarthritic temporomandibular joints acts as a lytic agent. PMID- 11500734 TI - Pneumoperitoneum and free meconium without gastrointestinal perforation in a neonate. AB - A newborn was referred to our hospital because of poor feeding and abdominal distension and was found to have pneumoperitoneum on abdominal x-ray. At operation there was free intraperitoneal air with no free fluid in the peritoneal cavity. In addition there was free air and meconium retroperitoneally on the left side but there was no evidence of gastrointestinal perforation. This case is rare and unique in that no demonstrable cause for the free air and free meconium could be demonstrated. PMID- 11500735 TI - Intussusception. A cause of postoperative intestinal obstruction in children. AB - Intestinal obstruction is a common postoperative complication and usually related to intra-abdominal adhesions. Postoperative intussusception, however, is a rare cause and may be confused with postoperative ileus. With more children undergoing abdominal operations, the incidence of postoperative intussusception should rise but reports indicate that this complication may be either not suspected or overlooked. We report 2 children who developed postoperative intussusception following repair of ruptured urinary bladder in one and appendicectomy in the other. Both had successful manual reduction at laparotomy. PMID- 11500736 TI - Long term intubation and successful weaning in two children with Guillain-Barre syndrome. AB - No definite criteria exists in Guillian-Barre syndrome in children regarding prolonged ventilation through an endo-tracheal tube without tracheostomy and successful weaning using a T-piece. Here we report two such cases of Guillian Barre syndrome requiring prolonged intubation for 56 days and ventilation for 30 days and ultimately successfully weaning them using the T-piece. Both the children eventually made a complete recovery, highlighting the point that in children prolonged intubation and ventilation using the portex tube is equally good, if not, better than tracheostomy with its attendant risks. PMID- 11500737 TI - Cutaneous myiasis due to Dermatobia hominis in Saudis. AB - Cutaneous myiasis infestations are normally found in South and Central America but increasing travel has resulted in their spread to non-indigenous countries with increasing frequency. We report two cases of cutaneous infestation by Dermatobia hominis in Taif, Saudi Arabia. There was no history of travel outside Saudi Arabia. The source of infection appears to be domestic cattle indicating that these infestations may be endemic in this region. PMID- 11500738 TI - Women's height age or age height index and risk of cesarean delivery. PMID- 11500739 TI - Neonatal varicella and acyclovir. PMID- 11500740 TI - Guidelines and code of ethics. PMID- 11500742 TI - Primary hyperparathyroidism. PMID- 11500743 TI - [Use of transesophageal echocardiography during implantation of aortic endoprosthesis (stent). Initial experience]. AB - OBJECTIVE: To report the role played by transesophageal echocardiography during implantation of self-expanding aortic endoprostheses (stent) at a hemodynamics laboratory. METHODS: Thirteen patients underwent stent implantation in the descending thoracic aorta with the aid of transesophageal echocardiography during the entire procedure. Indications for stenting were as follows: 8 aortic dissections, 2 true aneurysms, 2 penetrating atherosclerotic ulcers, and 1 traumatic pseudoaneurysm. RESULTS: No complications resulting from the use of transesophageal echocardiography were observed. In 12 patients, the initial result was considered appropriate, with total or partial resolution of the major lesion confirmed by a posterior examination. In 1 patient, the procedure was suspended after transesophageal echocardiography and angiography showed that the proximal aortic diameter was inappropriate. Transesophageal echocardiography contributed to clarifying relevant points, such as aortic diameter, anatomic detail of the intimal lesion, and location and size of the communicating orifice. In addition, it facilitated placing the stent in the target lesion, reduced the time of exposure to radiation and the use of contrast medium, and provided rapid identification of intercurrent events, possibly reducing the total duration of the procedure. CONCLUSION: The use of transesophageal echocardiography during placement of aortic stents seems appropriate. The actual advantages of the procedure will be defined in a comparative prospective study. PMID- 11500744 TI - Prevalence of hypertension in the urban population of Catanduva, in the State of Sao Paulo, Brazil. AB - OBJECTIVE: To study the prevalence of systemic hypertension and its control in the population of Catanduva, in the state of Sao Paulo, Brazil. METHODS: We carried out a randomized cross-sectional population-based study of the urban population of Catanduva with individuals above 18 years of age (688 individuals accounting for 0.9% of the referred population). We interviewed study participants to analyze the major qualitative and quantitative variables that could influence the hypertensive scenario and the risk for systemic hypertension. Blood pressure was measured through the indirect method according to the III Consenso Brasileiro de Hipertensao (III Brazilian Consensus on Hypertension), which established blood pressure levels >/= 140/90 mm Hg as hypertensive. RESULTS: The prevalence of systemic hypertension was higher in individuals with: (1) history of hypertension (p<0.0001); (2) diabetes mellitus (p=0.05); (3) body mass index (B. M. I) >/= 25 kg/m(2) (p<0.001); (4) low educational level (p<0.0001); (5) familial income ranging from 1 to 5 minimum wages (p<0.05); (6) unmarried status (divorced/separated and widow(er)s) (p<0.0001). Of the interviewed individuals, 27.6% (p=0.05) had blood pressure levels under control. CONCLUSION: Our study showed that the prevalence of systemic hypertension was 31.5%, and that 27.6% of the individuals interviewed had blood pressure levels under control at the time of the interview. PMID- 11500745 TI - Do cardiologists at a university hospital adopt the guidelines for the treatment of heart failure? AB - OBJECTIVE: To verify whether the guidelines for the treatment of heart failure have been adopted at a university hospital. The guidelines recommend the following: use of angiotensin-converting enzyme inhibitors for all patients with systolic ventricular dysfunction, use of digitalis and diuretics for symptomatic patients, use of beta-blockers for patients in functional classes II or III, use of spironolactone for patients in functional classes III or IV. METHODS: We analyzed the prescriptions of 199 patients. All these patients had ejection fraction (EF) 70 years (p<0.001), the presence of previous coronary artery disease (p=0.0004), previous myocardial infarction (p<0.001), infarction in the anterior wall (p=0.007), presence of left ventricular dysfunction (p<0.001), and the absence of thrombolytic therapy (p=0.04). Through the multivariate analysis of logistic regression, the following variables were associated with in-hospital mortality: female sex (p=0.001), age (p=0.008), the presence of previous myocardial infarction (p=0.02), and left ventricular dysfunction (p<0.001). CONCLUSION: After adjusting for all risk variables, female sex proved to be a variable independently related to in-hospital mortality in acute myocardial infarction. PMID- 11500747 TI - [Functional characterization of myocardial hypertrophy induced by isoproterenol and its regression]. PMID- 11500748 TI - Chagas' heart disease: evolutive evaluation of electrocardiographic and echocardiographic parameters in patients with the indeterminate form. AB - OBJECTIVE: To identify and associate potential electrocardiographic and echocardiographic changes in patients with the indeterminate form of Chagas' disease during long-term follow-up. METHODS: One hundred sixty patients underwent standard electrocardiography and two-dimensional guided M-mode echocardiography for left ventricular ejection fraction determination. Patients were followed up for 98.6+/-30.4 months, undergoing repeat electrocardiographic studies at 6-month intervals and echocardiographic studies at 12-month intervals. RESULTS: Based on the electrocardiographic findings, the patients were divided into group I, 125 patients (78.6%) with normal electrocardiograms throughout follow-up, and group II, 34 patients (21.3%) who developed electrocardiographic changes. Group II was further divided into group IIA (9 patients, 5.6%) with permanent electrocardiographic changes, group IIB (14 patients, 8.8%) with transitory electrocardiographic changes, and group IIC (11 patients, 6.9%) with changes appearing only on the final electrocardiogram. Left ventricular ejection fractions remained normal in the entire population studied and did not differ among groups. CONCLUSION: The indeterminate form of Chagas' disease clearly represents a benign condition with a favorable long-term prognosis. Although some patients develop electrocardiographic changes, left ventricular systolic function is well preserved. PMID- 11500749 TI - [Echocardiographic diagnosis of transposition of the great arteries associated with anomalous pulmonary venous connection]. AB - We report 2 cases of transposition of the great arteries associated with anomalous pulmonary venous connection emphasizing the clinical findings, the diagnosis, and the evolution of the association. One of the patients had the anomalous pulmonary venous connection in its total infradiaphragmatic form, in the portal system, and the other patient had a partial form, in which an anomalous connection of the left superior lobar vein with the innominate vein existed. At the time of hospital admission, the patients had cyanosis and respiratory distress with clinical findings suggesting transposition of the great arteries. The diagnosis in 1 of the cases, in which the anomalous connection was partial, was established only with echocardiography, without invasive procedures that would represent risk for the patient; in the other case, in which the anomalous connection was total, the malformation was only evidenced with catheterization. The patients underwent surgery for anatomical correction of the heart disease. Only 1 patient had a good outcome. PMID- 11500750 TI - Uhl's anomaly. Differential diagnosis and indication for cardiac transplantation in an infant. AB - We report the case of an 8-month-old female infant with Uhl's anomaly, who underwent successful cardiac transplantation. The clinical findings, complementary laboratory tests, anatomic findings, and differential diagnosis of the anomaly are discussed. PMID- 11500751 TI - Fibroelastoma of the mitral valve as a cause of transient ischemic stroke. AB - A 44-year-old woman had a transient ischemic stroke, fibroelastoma of the mitral valve being the source of the embolus. The patient evolved with neutropenia induced by ticlopidine after 10 days of treatment. We report the major clinical features, therapeutical options, and medicamentous toxicity resulting from the use of antiplatelet drugs. PMID- 11500752 TI - [Clinic-radiographic correlation. Case 7/2001 - Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 11500753 TI - [Use of intravascular ultrasonics in interventional Cardiology]. PMID- 11500754 TI - Vectorial competence of Aedes aegypti (Linnaeus 1762) Rio de Janeiro strain, to Dirofilaria immitis (Leidy 1856). AB - Dirofilaria immitis (Leidy 1856), a nematode parasite, is the etiologic agent of canine heartworm disease and mosquitoes are essential intermediate hosts. Mosquito susceptibility to the worms differ with species, strains and also among individuals of the same strain. To evaluate the degree of susceptibility of Rio de Janeiro laboratory raised strain of Aedes aegypti, we fed mosquitoes on canine blood with different densities of microfilariae (mf). There was no significant difference in the rate of development among the three different densities of mf. Infective larvae were found in the head and proboscis of all mosquitoes provided bloodmeals with different densities of mf after the 11th day post-infection. The infection rate of mosquitoes after ingestion of blood containing 3,000 mf/ml, 5,000 mf/ml and 7,000 mf/ml were 55.3%, 66.7% and 100%, respectively. The vector efficiency indices ranged from 1.6 to 9.3. The finding of L3 stage larvae, high infection rates and vector efficiency indices suggest that Ae. aegypti, Rio de Janeiro laboratory strain, is a potential vector of D. immitis, although of low efficiency. PMID- 11500755 TI - Trypanosoma evansi control and horse mortality in the Brazilian Pantanal. AB - The impact of three treatment strategies for Trypanosoma evansi control on horse mortality in the Brazilian Pantanal based on four size categories of cattle ranches is explored. The region's 49,000 horses are indispensable to traditional extensive cattle ranching and T. evansi kills horses. About 13% of these horses would be lost, annually, due to T. evansi if no control were undertaken. One preventive and two curative treatment strategies are financially justifiable in the Pantanal. The best available technology for the treatment of T. evansi from a horse mortality perspective is the preventive strategy, which spares 6,462 horses, annually. The year-round cure spares 5,783 horses, and the seasonal cure saves 5,204 horses on a regional basis relative to no control strategy. Regardless of the strategy adopted, 39% of the costs or benefits fall to the largest ranches, while 18% fall to the smallest ranches. PMID- 11500756 TI - Small rodents fleas from the bubonic plague focus located in the Serra dos Orgaos Mountain Range, State of Rio de Janeiro, Brazil. AB - Eleven species of fleas were collected from 601 small rodents, from November 1995 to October 1997, in areas of natural focus of bubonic plague, including the municipalities of Nova Friburgo, Sumidouro and Teresopolis, State of Rio de Janeiro, Brazil. Among 924 fleas collected, Polygenis (Polygenis) rimatus (Rhopalopsyllidae) was the predominant species regarding the frequency, representing 41.3% (N:382), followed by P. (Neopolygenis) pradoi, representing 20% (N:185) and Craneopsylla minerva minerva (Stephanocircidae), representing 18.9% (N:175). The host Akodon cursor harbored 47.9% of these fleas. Other six host species were infested by 52.1% of the remaining fleas. Fleas were found on hosts and in places within the focus not previously reported by the literature. PMID- 11500757 TI - Biogeography of Triatominae (Hemiptera: Reduviidae) in Ecuador: implications for the design of control strategies. AB - Chagas disease control strategies strongly depend on the triatomine vector species involved in Trypanosoma cruzi transmission within each area. Here we report the results of the identification of specimens belonging to various species of Triatominae captured in Ecuador (15 species from 17 provinces) and deposited in the entomological collections of the Catholic University of Ecuador (Quito), Instituto Oswaldo Cruz (Brazil), the Natural History Museum London (UK), the London School of Hygiene and Tropical Medicine (UK), the National Institute of Hygiene (Quito), and the Vozandes Hospital (Quito). A critical review of published information and new field records are presented. We analysed these data in relation to the life zones where triatomines occur (11 life zones, excluding those over 2,200 m altitude), and provide biogeographical maps for each species. These records are discussed in terms of epidemiological significance and design of control strategies. Findings relevant to the control of the main vector species are emphasised. Different lines of evidence suggest that Triatoma dimidiata is not native to Ecuador-Peru, and that synanthropic populations of Rhodnius ecuadoriensis in southern Ecuador-northern Peru might be isolated from their sylvatic conspecifics. Local eradication of T. dimidiata and these R. ecuadoriensis populations might therefore be attainable. However, the presence of a wide variety of native species indicates the necessity for a strong longitudinal surveillance system. PMID- 11500758 TI - Enteropathogens associated with diarrheal disease in infants of poor urban areas of Porto Velho, Rondonia: a preliminary study. AB - One hundred and thirty cases of diarrhea and 43 age-matched controls, 0 to 5 years old, were studied in a pediatric outpatient unit from a poor peri urban area of Porto Velho, Rondonia. Eighty percent of diarrheal cases were observed in the groups under 2 years of age. Rotavirus (19.2%) was the most frequent enteropathogen associated with diarrhea, followed by Shigella flexneri (6.15%) and S. sonnei (1.5%) and Salmonella sp. (6.9%). Four cases of E. coli enterotoxigenic infections (3.1%), E. coli enteropathogenic (EPEC)(2.3%) one case of E. coli enteroinvasive infection (0.8%) and one case of Yersinia enterocolitica (0.8%) were also identified. Mixed infections were frequent, associating rotavirus, EPEC and Salmonella sp. with Entamoeba histolytica and Giardia lamblia. PMID- 11500759 TI - Spatio-temporal dynamics and transition from asymptotic equilibrium to bounded oscillations in Chrysomya albiceps (Diptera, Calliphoridae). AB - The sensitivity of parameters that govern the stability of population size in Chrysomya albiceps and describe its spatial dynamics was evaluated in this study. The dynamics was modeled using a density-dependent model of population growth. Our simulations show that variation in fecundity and mainly in survival has marked effect on the dynamics and indicates the possibility of transitions from one-point equilibrium to bounded oscillations. C. albiceps exhibits a two-point limit cycle, but the introduction of diffusive dispersal induces an evident qualitative shift from two-point limit cycle to a one fixed-point dynamics. Population dynamics of C. albiceps is here compared to dynamics of Cochliomyia macellaria, C. megacephala and C. putoria. PMID- 11500760 TI - Hepatitis B infection among patients attending a sexually transmitted diseases clinic in Rio de Janeiro, Brazil. AB - Hepatitis B virus (HBV) has a low endemicity in Rio de Janeiro, Brazil. Sexual transmission must play an important role in this virus, but the prevalence and risk factors have never been properly investigated. The aim of this paper is to determine the prevalence and risk factors for HBV infection in patients attending a Sexually Transmitted Diseases Clinic of the Universidade Federal Fluminense, from the State of Rio de Janeiro, Brazil. In a retrospective study, HBV seroprevalence was investigated in 440 patients. Serum of each patient was assayed for antibodies against hepatitis B core antigen (anti-HBc), hepatitis B surface antigen (HBsAg) and antibodies against hepatitis B surface antigen (anti HBs). Demographic and risk factor data were extracted from clinic notes. The overall seroprevalence of exposure markers for HBV (anti-HBc, HBsAg and anti-HBs) were 13%, 3.4% and 8.5% respectively. Homo/bisexual behaviour, anal intercourse, HIV infection, positive serology for syphilis and blood transfusion were predictors of the HBV exposure. Among demographic data, age and place of birth were associated with the anti-HBc seropositivity. PMID- 11500761 TI - Attractiveness of black Shannon trap for phlebotomines. AB - A white Shannon-type trap was used for captures of female sand flies in the search for natural infection with flagellates, however, due to its low productivity and as a large number of phlebotomines settled on the researchers' black clothes, we decided to compare the relative attractiveness of black and white Shannon-type traps for sand flies. Several pairs of black and white traps were placed side by side in front of caves in four areas in the Serra da Bodoquena, Bonito county, State of Mato Grosso do Sul, Brazil, for a total of 12 observations and 44 h of capture. The experiment resulted in 889 phlebotomines captured, 801 on the black and 88 on the white trap, representing 13 species. The hourly Williams' means were 8.67 and 1.24, respectively, and the black/white ratio was 7.0:1.0. Lutzomyia almerioi, an anthropophilic species closely associated with caves, was predominant (89%). Only two other species, Nyssomyia whitmani and Psathyromyia punctigeniculata, also anthropophilic, were significantly attracted to the black rather than to the white trap (chi(2) test; p < or = 0.01). The difference between the diversity index of the two traps was not significant at level 0.05. The black trap in these circumstances was much more productive than the white, especially for anthropophilic species. PMID- 11500762 TI - Force of infection and evolution of lesions of canine tegumentary leishmaniasis in northwestern Argentina. AB - A clinical-serological follow-up was carried out in a canine population in endemic foci of Leishmania braziliensis spread in northwestern Argentina. Each dog was studied in at least two visits, 309+/-15 days (X+/-SE) apart. Some initially healthy dogs (n=52) developed seroconversion or lesions. The clinical evolution of the disease in dogs resembles in many aspects the human disease. Similarities include the long duration of most ulcers with occasional healing or appearance of new ones and the late appearance of erosive snout lesions in some animals. Yearly incidence rates of 22.7% for seroconversion and of 13.5% for disease were calculated as indicators of the force of infection by this parasite upon the canine population. PMID- 11500763 TI - Human infection by Pseudoterranova decipiens (Nematoda, Anisakidae) in Chile: report of seven cases. AB - From 1997 to 1999, we identified seven human cases of infection by fourth stage larvae of Pseudoterranova decipiens in Chile. All identified larvae were coughed up by the patients. Subjects were 10-55 years old; five were female. Some patients complained of coughing, expectoration, pharyngeal pain, nausea or anal and nasal pruritus. Larvae of three patients were coughed up from 36 h to 7 days after having eaten raw (cebiche or sushi) or lightly fried fish. P. decipiens has a marine life cycle. Infective third stage larva develop to adult stage in pinniped mammals. The nematode eggs are voided with the host faeces and develop and hatch releasing third stage larvae. Some crustaceans and fish act as hosts of third stage larvae. Man is an accidental host for third or fourth stage larvae. PMID- 11500764 TI - Reappearance of Aedes aegypti (Diptera: Culicidae) in Lima, Peru. AB - We report here the reappearance of Aedes aegypti in the Rimac district, and summarize the history of this mosquito species in Peru since its first detection in 1852. On March 17 2000 were found Ae. aegypti and Culex quinquefasciatus in Mariscal Castilla town, Flor de Amancaes, San Juan de Amancaes, El Altillo and Santa Rosa in the Rimac district, Lima Province. PMID- 11500765 TI - Evidence of colonization of man-made ecotopes by Triatoma dimidiata (Latreille, 1811) in Costa Rica. AB - Triatoma dimidiata adults have been frequently found, during the last five years, in a dog kennel and a chicken coop, in the back yard of a well-built house, 15 km from San Jose, the capital of Costa Rica. In the chicken coop nymphs were also found. Two of the 11 dogs from the kennel were serologically positive for Trypanosoma cruzi infection. The inhabitants of the house, three adults and two children, were negative. This type of colonization by the insect, which is attracted to lights, is becoming common in old and new settlements, with different degrees of success, a fact with epidemiological implications and great relevance in the control strategies that can be applied. PMID- 11500766 TI - Identification of Biomphalaria havanensis and Biomphalaria obstructa populations from Cuba using polymerase chain reaction and restriction fragment length polymorphism of the ribosomal RNA intergenic spacer. AB - In Cuba, several Biomphalaria species have been reported such as B. orbignyi, B. schrammi, B. helophila, B. havanensis and B. peregrina; only the latter three are considered as potential hosts of Schistosoma mansoni. The specific identification of Biomphalaria species is based on anatomical and morphological characters of genital organs and shells. The correct identification of these snails is complicated by the high variation in these characters, similarity among species and in some cases by the small size of the snails. In this paper, we reported the classical morphological identification, the use of PCR and RFLP analysis of the internal transcribed spacer region of the ribosomal RNA genes for molecular identification of seven snail populations from different localities in Cuba. Using morphological and molecular analysis, we showed that among the studied Cuban Biomphalaria populations only B. havanensis and B. obstructa species were found. PMID- 11500767 TI - New host records and description of the egg of Anacanthorus penilabiatus (Monogenea, Dactylogyridae). AB - Anacanthorus penilabiatus is referred parasitizing the type-host Piaractus mesopotamicus (Serrasalmidae) and two new hosts, Colossoma macropomum and C. brachypomum (Characidae) from fish ponds of "Departamento Nacional de Obras Contra as Secas", Pentecoste, State of Ceara, Brazil. Table of measurements and the first description of the egg are presented. PMID- 11500768 TI - Immunological follow-up of hydatid cyst cases. AB - Hydatid disease is caused by Echinococcus granulosus. In this study, we aimed to investigate the benefit of monitoring cases with hydatid cyst by means of immune components in patients in a long-term follow-up after surgery. Eighty-four preoperative and postoperative serum samples from 14 cases undergoing surgery for hydatid disease were evaluated in terms of immune parameters, such as total and specific IgE, IgG, IgM, IgA and complement. Total and specific IgE were determined by ELISA. Specific IgG levels were measured by indirect hemagglutination. Total IgG, IgM, IgA and complement (C3 and C4) were detected by nephelometry. Imaging studies were also carried out during the follow-up. In none of the patients hydatid cysts were detected during the follow-up. Total IgE levels in the sera of the patients decreased to normal six months after surgery. Although specific IgE against echinococcal antigens decreased one year after operation, levels were still significantly high. There were no changes in the levels of anti-Echinococcus IgG and total IgG in follow-up period. Additionally, other parameters, such as IgA, IgM, C3 and C4, were not affected. PMID- 11500769 TI - In vivo lymphocyte activation and apoptosis by lectins of the Diocleinae subtribe. AB - This paper reports the overall effects of three lectins, extracted from Canavalia brasiliensis, Dioclea violacea, and D. grandiflora, on BALB/c mice popliteal draining lymph nodes. These lectins have presented high stimulatory capacity on lymph node T cells. Additionally, they were able to induce apoptosis and inflammation (frequently associated with high endothelial venule necrosis). The data presented here suggest that the Diocleinae lectins studied can stimulate in vivo T cell activation and apoptosis, as well as present important side effects. PMID- 11500770 TI - Skin reactions to thimerosal and Leishmania in dogs from a leishmaniasis endemic area: it is better to keep them apart. AB - Positive Montenegro's skin test is a delayed type hypersensitivity reaction widely used as indicative of previous infection with Leishmania in both humans and dogs. Montenegro's antigen consists of a crude Leishmania antigen solution, usually containing thimerosal as preserving agent. In this work it is shown that a large proportion of dogs (11 out of 56) examined in an endemic area of leishmaniasis presented induration at the site of injection of a diluent containing thimerosal alone. This clearly demonstrates that thimerosal leads to a high number of false positive skin reactions in dogs and that its use in Montenegro's skin test antigenic preparations should be avoided. PMID- 11500771 TI - Diagnostic performance characteristics of rapid dipstick test for Plasmodium vivax malaria. AB - We compared the diagnostic performance characteristics of newly developed method, the rapid dipstick test, which provides colorimetric determination by developing antibody to the lactate dehydrogenase enzyme of parasites, with conventional standard thick-blood film examination. For the rapid test, OptiMAL commercial kits were used. The results were also evaluated with clinical findings from patients. The parasites were determined by microscopic examination of thick-blood films from 81 patients with vivax malaria from southeastern Anatolia, Turkey. The OptiMAL test results were found to be negative in five patients who were diagnosed clinically and through thick-film testing as having vivax malaria. There was no false positivity observed with the OptiMAL test. We concluded that this rapid malaria test has a lower level of sensitivity than the classical thick blood-film test for malaria, but that these methods have equal specificity. PMID- 11500772 TI - Seroprevalence of Toxoplasma gondii infection in goats by the indirect haemagglutination, immunofluorescence and immunoenzymatic tests in the region of Uberlandia, Brazil. AB - A comparative study of the indirect haemagglutination (IHA), immunofluorescence (IFAT) and immunoenzymatic (ELISA) tests was carried out to determine the prevalence of Toxoplasma gondii antibodies in goats. One hundred seventy-four serum samples were obtained from four goat herds from the region of Uberlandia, State of Minas Gerais. The distribution of the animals, according to their origin, was as follow: 71 from herd I; 39 from herd II; 37 from herd III; and 27 from herd IV. Serum samples were analyzed by IHA, IFAT and ELISA, considering the reactivity of the serum samples at dilution > or = 1:64 as cut off titer for the three tests. A global seroprevalence of 18.4% was observed, with significantly higher positivity rate in the herd II (66.7%) and older animals (> 36 months). A high and significant positive correlation was found between the titers obtained by the IHA versus IFAT, IHA versus ELISA, and ELISA versus IFAT. Therefore, it can be concluded that the three analyzed tests have shown to be highly concordant and appropriate for epidemiological surveys of Toxoplasma infection in goats. Although the seroprevalence of T. gondii infection in goats is relatively low in this region as compared to other regions of the country, adequate management might be useful and essential to control the infection in the goat herds. PMID- 11500773 TI - Comparison between precipitin and ELISA tests in the bloodmeal detection of Aedes aegypti (Linnaeus) and Aedes fluviatilis (Lutz) mosquitoes experimentally fed on feline, canine and human hosts. AB - The identification of arthropod bloodmeals is important in many epidemiological studies, as, the understanding of the life cycle of vectors and the pathogens they transmit, as well as helping to define arthropods' control strategies. The precipitin test has been used for decades, but ELISA is slowly becoming more popular. To compare the two tests for sensitivity, specificity and accuracy to detect small insect bloodmeals, Aedes aegypti or Ae. fluviatilis mosquitoes were fed either on feline, canine or human hosts. Mosquitoes were frozen at 6, 12, 24, 48 or 72 h after feeding. Precipitin test showed better specificity and accuracy and ELISA test showed higher sensitivity. Better results with both tests were achieved when mosquitoes were frozen within 48 h from feeding. PMID- 11500774 TI - A alpha-glycerophosphate dehydrogenase is present in Trypanosoma cruzi glycosomes. AB - alpha-glycerophosphate dehydrogenase (alpha-GPDH-EC.1.1.1.8) has been considered absent in Trypanosoma cruzi in contradiction with all other studied trypanosomatids. After observing that the sole malate dehydrogenase can not maintain the intraglycosomal redox balance, GPDH activity was looked for and found, although in very variable levels, in epimastigotes extracts. GPDH was shown to be exclusively located in the glycosome of T. cruzi by digitonin treatment and isopycnic centrifugation. Antibody against T. brucei GPDH showed that this enzyme seemed to be present in an essentially inactive form at the beginning of the epimastigotes growth. GPDH is apparently linked to a salicylhydroxmic-sensitive glycerophosphate reoxidizing system and plays an essential role in the glycosome redox balance. PMID- 11500775 TI - Production of cytolethal distending toxin and other virulence characteristics of Escherichia coli strains of serogroup O86. AB - Genetic and phenotypic virulence markers of different categories of diarrhoeagenic Escherichia coli were investigated in 106 strains of enteropathogenic E. coli (EPEC) serogroup O86. The most frequent serotype found was O86:H34 (86%). Strains of this serotype and the non motile ones behaved as EPEC i.e., carried eae, bfpA and EAF DNA sequences and presented localised adherence to HeLa cells. Serotypes O86:H2, O86:H6, O86:H10, O86:H18, O86:H27 and O86:H non determined, belonged to other categories. The majority of the strains of serotype O86:H34 and non motile strains produced cytolethal-distending toxin (CDT). The ribotyping analysis showed a correlation among ribotypes, virulence markers and serotypes, thus suggesting that CDT production might be a property associated with a universal clone represented by the O86:H34 serotype. PMID- 11500776 TI - Variants of the Plasmodium vivax circumsporozoite protein (VK210 and VK247) in Colombian isolates. AB - Phenotypic diversity has been described in the central repeated region of the circumsporozoite protein (CSP) from Plasmodium vivax. Two sequences VK210 (common) and VK247 (variant) have been found widely distributed in P. vivax isolates from several malaria endemic areas around the world. A third protein variant called P. vivax-like showing a sequence similar to the simian parasite P. simioovale has also been described. Here, using an immunofluorescent test and specific monoclonal antibodies, we assessed the presence of two of these protein variants (VK210 and VK247) in laboratory produced sporozoite. Both sequences were found in parasite isolates coming from different geographic regions of Colombia. Interestingly, sporozoites carrying the VK247 sequence were more frequently produced in Anopheles albimanus than sporozoites with the VK210 sequence. This difference in sporozoites production was statistically significant (p <0.05, Kruskal-Wallis); not correlation was found with parameters as the total number of parasites or gametocytes in blood from human donors used to feed mosquitoes. Previous studies in the same region have shown a higher prevalence of anti-VK210 antibodies which in theory may suggest their role in blocking the development of sporozoites carrying the CSP VK210 sequence. PMID- 11500777 TI - Experimental transmission of Trypanosoma cruzi through the genitalia of albino mice. AB - Trypanosoma cruzi is usually transmitted by contact with the excreta of infected Triatominae; among non-vectorial infections, direct transmission through coitus has been proposed. We investigated this possibility by instilling, through the external meatus of the vagina and the penis of previously anesthetized NMRI albino mice, blood of mice infected with strains isolated from Didelphis marsupialis (opossum, strain CO57), Rattus rattus (rat, strain CO22) and human (strain EP). Some animals were allowed to copulate the same day of the instillation. In other experiments, the strains were inoculated in the scrotum. To determine the effect of immunosuppression, some mice were treated with cyclophosphamide 30 days post-instillation. Controls were instilled orally and ocularly. Vaginal instillation with strain CO22 produced systemic infection with tropism to the heart, skeletal muscle, skin, duodenum, pancreas, ovary and sternum. Scrotal inoculation with strain EP likewise invaded liver, spleen, lung, lymph nodes and urogenital organs; while strain CO57 invaded skeletal and cardiac muscle, pancreas, testis, and vas deferens. Penile infection with strain CO22 was detected by xenodiagnosis. Immunosuppression did not increase parasitemia of vaginally infected mice or controls. Mating did not produce infection. Our results show that contact of blood trypomastigotes of T. cruzi with genital mucosa can produce blood and tissue infections. These results are discussed in relation to reports of frequent experimental tropism of T. cruzi toward urogenital organs. PMID- 11500778 TI - Sequential histological changes in Biomphalaria glabrata during the course of Schistosoma mansoni infection. AB - Biomphalaria glabrata, highly susceptible to Schistosoma mansoni, were seen to shed less and less cercariae along the time of infection. Histological examination kept a close correlation with this changing pattern of cercarial shedding, turning an initial picture of no-reaction (tolerance) gradually into one of hemocyte proliferation with formation of focal encapsulating lesions around disintegrating sporocysts and cercariae, a change that became disseminated toward the 142nd day post miracidial exposure. Findings were suggestive of a gradual installation of acquired immunity in snails infected with S. mansoni. PMID- 11500779 TI - Biological activities of Curcuma longa L. AB - There are several data in the literature indicating a great variety of pharmacological activities of Curcuma longa L. (Zingiberaceae), which exhibit anti-inflammatory, anti-human immunodeficiency virus, anti-bacteria, antioxidant effects and nematocidal activities. Curcumin is a major component in Curcuma longa L., being responsible for its biological actions. Other extracts of this plant has been showing potency too. In vitro, curcumin exhibits anti-parasitic, antispasmodic, anti-inflammatory and gastrointestinal effects; and also inhibits carcinogenesis and cancer growth. In vivo, there are experiments showing the anti parasitic, anti-inflammatory potency of curcumin and extracts of C. longa L. by parenteral and oral application in animal models. In this present work we make an overview of the pharmacological activities of C. longa L., showing its importance. PMID- 11500780 TI - Resistance of Musca domestica L. populations to cyromazine (insect growth regulator) in Brazil. AB - Five field populations of Musca domestica L. collected in poultry farms were bioassayed in order to detect possible resistance to the larvicide cyromazine in Brazil. The concentrations used were 0, 0.5, 0.1, 0.2, 0.4, 1, 2, 4 and 8 ppm. Three populations (Petropolis, RJ, Montes Claros, MG and Promissao, SP) were resistant, while the other two populations (Ibiuna, SP and Monte Mor, SP) were more susceptible than the reference pathern used by the World Health Organization. The presence of three resistant house fly populations to cyromazine in Brazilian poultry farms strongly suggests that the operational aspects of larvicide use are important for the resistance development. Cyromazine is applied as a feed-through, both in Brazil and in the USA, where resistance has already been documented. However, in Denmark, where it was approved only as a topical manure spray, no case of resistance has yet been detected. PMID- 11500781 TI - Seasonal analysis of the number of aeropiles in Anocentor nitens (Neumann, 1897) (Acari: Ixodidae) from the State of Rio de Janeiro, Brazil. AB - Variation between aeropile numbers of the right and left peritrematic plate in male and female Anocentor nitens (Neumann, 1897) is reported from a site in Brazil. From January to December 1998, 146 males and 247 females of A. nitens were recovered from Equus caballus L. in Silva Jardim District, State of Rio de Janeiro. Asymmetry of numbers of aeropiles between right and left plates occurred in 83.6% of the males and 82.2% of the females. Differences in the number of aeropiles between the sexes were not significant. Quantitative variation of aeropiles was correlated to the period of recovery, with significant asymmetry detected in August-September and November-December, mainly in males. Results suggest an adaptation, especially in the male ticks, that expresses itself as greater variation in the number of aeropiles in some periods of the year. PMID- 11500782 TI - Effects of the desiccation on Biomphalaria tenagophila (Orbigny, 1835) (Mollusca) infected by Schistosoma mansoni Sambon, 1907. AB - Specimens of Biomphalaria tenagophila exposed to miracidia of Schistosoma mansoni were submitted to different desiccation periods as follows: group I: 24 h after exposure, desiccated for 28 days; group II: after cercariae elimination, desiccated for 7 days; group III: 21 days after exposure, desiccated for 7 days; group IV: 14 days after exposure, desiccated for 14 days; group V: 7 days after exposure, desiccated for 21 days. From the obtained data it was verified that desiccation was not capable of interrupting the development of larvae of S. mansoni in mollusks. A delay in the development of S. mansoni larvae in groups I, III, IV and V was observed. A pause was verified in the development of S. mansoni larvae in groups II, III, IV and V. Some larvae, in groups I, III, IV and V, did not suffer as a result of desiccation and continued their development. Larvae in the cercariae stage were shown to be more sensitive to desiccation. It was possible to obtain clearing of mollusks infected by sporocysts II and cercariae using a period of 7 days of desiccation. PMID- 11500783 TI - Salt and Brazilian ancestry. PMID- 11500785 TI - Iron deficiency in blood donors. AB - CONTEXT: Blood donation results in a substantial loss of iron (200 to 250 mg) at each bleeding procedure (425 to 475 ml) and subsequent mobilization of iron from body stores. Recent reports have shown that body iron reserves generally are small and iron depletion is more frequent in blood donors than in non-donors. OBJECTIVE: The aim of this study was to evaluate the frequency of iron deficiency in blood donors and to establish the frequency of iron deficiency in blood donors according to sex, whether they were first-time or multi-time donors, and the frequency of donations per year. DESIGN: From September 20 to October 5, 1999, three hundred blood donors from Santa Casa Hemocenter of Sao Paulo were studied. DIAGNOSTIC TESTS: Using a combination of biochemical measurements of iron status: serum iron, total iron-binding capacity, transferrin saturation index, serum ferritin and the erythrocyte indices. RESULTS: The frequency of iron deficiency in blood donors was 11.0%, of whom 5.5% (13/237) were male and 31.7% (20/63) female donors. The frequency of iron deficiency was higher in multi-time blood donors than in first-time blood donors, for male blood donors (7.6% versus 0.0%, P < 0.05) and female ones (41.5% versus 18.5%, P < 0.05). The frequency of iron deficiency found was higher among the male blood donors with three or more donations per year (P < 0.05) and among the female blood donors with two or more donations per year (P < 0.05). CONCLUSIONS: We conclude that blood donation is a very important factor for iron deficiency in blood donors, particularly in multi time donors and especially in female donors. The high frequency of blood donors with iron deficiency found in this study suggests a need for a more accurate laboratory trial, as hemoglobin or hematocrit measurement alone is not sufficient for detecting and excluding blood donors with iron deficiency without anemia. PMID- 11500786 TI - Intracavernous injection in the treatment of erectile dysfunction after radical prostatectomy: an observational study. AB - CONTEXT: Despite the recent improvements in performing radical retropubic prostatectomy that have led to a considerable decrease in the complication rate, erectile dysfunction still represents a major problem. Moreover, less invasive treatment options that are emerging for erectile dysfunction have not shown satisfactory results in managing these patients. OBJECTIVE: To study the efficacy and side effects of self-injection therapy in the treatment of men who had become impotent after undergoing radical prostatectomy due to prostate cancer, over a study period of 96 months. DESIGN: Observational study. SETTING: University Referral Center. PARTICIPANTS: 168 patients with erectile dysfunction, aged 43 to 78 years old, who underwent radical retropubic prostatectomy due to localized prostate cancer. PROCEDURES: The patients were treated with self-injection therapy using papaverine, phentolamine and prostaglandin E1, at home. RESULTS: This study showed an acceptable 94.6% success rate, with no life-threatening complications. In addition to this, our series presented a 13.1% cure rate with this therapy. CONCLUSION: Self-injection therapy with papaverine, phentolamine and prostaglandin E1 is effective and safe in the treatment of erectile dysfunction after radical prostatectomy. PMID- 11500787 TI - Comparison of positivity frequency of bcl-2 expression in prostate adenocarcinoma with low and high Gleason score. AB - CONTEXT: Multiple genetic and epigenetic factors have been implicated in the oncogenesis and progression of prostate cancer. The major difficulty is in that the clinical management stems from the reality that reliable and accurate prognostic biomarkers are not available and that effective treatment regimens forming hormone-resistant prostate cancers are yet to be developed. Among the most important regulators of apoptosis and programmed cell death is the bcl-2 gene and its related proteins. Elevated levels of bcl-2 protein may contribute to the progression of prostate cancers to a metastatic and hormone-insensitive state characterized by poor responses to chemotherapy. OBJECTIVE: To characterize the expression of bcl-2 proteins as a prognostic factor in humans. DESIGN: A retrospective approach. SETTING: Urology section, Federal University of Sao Paulo. DIAGNOSTIC TEST USED: Immunohistochemical analysis using bcl-2 protein antibody and normal staining by hematoxylin-eosin. MAIN MEASUREMENTS: Prognostic relations and protein expression were evaluated considering the total sample (28) divided into two groups, high (8 to 10) and low (2 to 4), separated according to the histological differentiation grade (Gleason score) with 10 and 18 samples, respectively. RESULTS: The differentiation of grade into two groups separated according to the Gleason score in low and high types presented different bcl-2 expression (P < 0.001). CONCLUSION: The higher frequency of bcl-2 immunostaining in tumor samples was observed in association with more advanced Gleason scores and suggests that an increase in the ratio of this anti-apoptotic protein often occurs during progression of prostate cancers. PMID- 11500788 TI - A 12-month follow-up of an influenza vaccination campaign based on voluntary adherence: report on upper-respiratory symptoms among volunteers and non volunteers. AB - CONTEXT: Routine immunization of groups at high risk for influenza has been progressively implemented as a matter of Brazilian public health policy. Although the benefits of the vaccination for healthy young adults are still controversial, it has been offered yearly to hundreds of thousands of Brazilian workers, generally as part of wellness initiatives in the workplace. OBJECTIVE: To study the characteristics of subjects that accepted or refused to be vaccinated against influenza and to report on respiratory symptoms in both groups, one year after the campaign date. DESIGN: A prospective observational study. SETTING: Workers at a subsidiary of an international bank in Sao Paulo, Brazil. PARTICIPANTS: 124 persons that did not accept and 145 that voluntarily accepted the vaccine completed 12 months of follow-up. MAIN MEASUREMENTS: Data concerning gender, age, tobacco use, and any history of chronic respiratory illness such as asthma, bronchitis, rhinitis, and repetitive upper-respiratory infections, were recorded at the time of vaccination. After that, workers were asked monthly by questionnaire or telephone about respiratory symptoms, days of work lost and medical consultations. RESULTS: The results showed statistically significant differences regarding age (P = 0.004) with the vaccinated group (V) being younger than the non-vaccinated (NV) one, and with reference to previous repetitive upper respiratory infections being higher among the V group (P < 0.0001). During the follow-up, the V group reported more occurrences of upper respiratory symptoms (P < 0.0001), due to both non-influenza (P < 0.0001) and influenza-like illness (P = 0.045). Differences were also found between V and NV groups concerning days off work and number of medical consultations due to upper-respiratory symptoms and non-influenza illness. Gender and history of repetitive upper-respiratory infections were the best predictors of influenza-like illness-related events. CONCLUSIONS: The making of previous reference to repetitive upper-respiratory infections was a major difference between those who accepted or rejected the vaccine. The vaccination itself was not sufficient to reduce the number of occurrences of respiratory symptoms and related absenteeism to levels similar to those found among non-vaccinated people. PMID- 11500789 TI - The frequency of Tay-Sachs disease causing mutations in the Brazilian Jewish population justifies a carrier screening program. AB - CONTEXT: Tay-Sachs disease is an autosomal recessive disease characterized by progressive neurologic degeneration, fatal in early childhood. In the Ashkenazi Jewish population the disease incidence is about 1 in every 3,500 newborns and the carrier frequency is 1 in every 29 individuals. Carrier screening programs for Tay-Sachs disease have reduced disease incidence by 90% in high-risk populations in several countries. The Brazilian Jewish population is estimated at 90,000 individuals. Currently, there is no screening program for Tay-Sachs disease in this population. OBJECTIVE: To evaluate the importance of a Tay-Sachs disease carrier screening program in the Brazilian Jewish population by determining the frequency of heterozygotes and the acceptance of the program by the community. SETTING: Laboratory of Molecular Genetics--Institute of Biosciences--Universidade de Sao Paulo. PARTICIPANTS: 581 senior students from selected Jewish high schools. PROCEDURE: Molecular analysis of Tay-Sachs disease causing mutations by PCR amplification of genomic DNA, followed by restriction enzyme digestion. RESULTS: Among 581 students that attended educational classes, 404 (70%) elected to be tested for Tay-Sachs disease mutations. Of these, approximately 65% were of Ashkenazi Jewish origin. Eight carriers were detected corresponding to a carrier frequency of 1 in every 33 individuals in the Ashkenazi Jewish fraction of the sample. CONCLUSION: The frequency of Tay-Sachs disease carriers among the Ashkenazi Jewish population of Brazil is similar to that of other countries where carrier screening programs have led to a significant decrease in disease incidence. Therefore, it is justifiable to implement a Tay-Sachs disease carrier screening program for the Brazilian Jewish population. PMID- 11500790 TI - Primary aortoenteric fistula related to septic aortitis. AB - CONTEXT: Primary aortoenteric fistulas usually result from erosion of the bowel wall due to an associated abdominal aortic aneurysm. A few patients have been described with other etiologies such as pseudoaneurysm originating from septic aortitis caused by Salmonella. OBJECTIVE: To present a rare clinical case of pseudoaneurysm caused by septic aortitis that evolved into an aortoenteric fistula. CASE REPORT: A 65-year-old woman was admitted with Salmonella bacteremia that evolved to septic aortitis. An aortic pseudoaneurysm secondary to the aortitis had eroded the transition between duodenum and jejunum, and an aortoenteric fistula was formed. In the operating room, the affected aorta and intestinal area were excised and an intestine-to-intestine anastomosis was performed. The aorta was sutured and an axillofemoral bypass was carried out. In the intensive care unit, the patient had a cardiac arrest that evolved to death. PMID- 11500791 TI - Breast cancer with choriocarcinomatous and neuroendocrine features. AB - CONTEXT: Breast cancer may express the presence of b-human chorionic gonadotrophin in 12% to 18% of cases, using immunohistochemical reactions. Usually the tumors will show positivity in a few scattered cells. Breast cancer with choriocarcinomatous features, as reported by Saigo and Rosen, is a distinct variant of breast cancer. We report a case of breast cancer with choriocarcinomatous and neuroendocrine features. OBJECTIVE: This is a case report of an invasive ductal carcinoma of the breast with choriocarcinomatous and neuroendocrine features. DESIGN: Case Report. CASE REPORT: A 50-year-old Brazilian woman underwent surgery for a lump in the right breast, which had been first noticed about 3 months earlier. The surgery consisted of quadrantectomy followed by right mastectomy with ipsilateral axillary lymph node dissection. The specimen from the quadrantectomy revealed a 7 x 6.5 x 4.5 cm tumor. Histology of the lesion showed the presence of an invasive ductal carcinoma with areas of giant cells and intense atypia. The immunohistochemistry was positive in the pleomorphic areas for human chorionic gonadotrophin, while the less pleomorphic areas showed positivity for synaptophysin and chromogranin. PMID- 11500792 TI - Convergence of the horizontal semicircular canal and otolith afferents on cat single vestibular neurons. AB - We studied the convergence of two afferent pairs of single vestibular neurons by selective stimulation of the horizontal semicircular canal (HC) and saccular (SAC) nerves, and the HC and utricular (UT) nerves in decerebrate cats. All recorded neurons were classified as vestibulospinal (VS), vestibulo-oculospinal (VOS) or vestibulo-ocular (VO), by antidromic stimulation from the oculomotor/trochlear nuclei and the spinal cord: neurons that could not be activated from any test sites were classified as vestibular (V) neurons. Of a total of 125 neurons activated by stimulation of the HC/SAC nerves, 21(17%) received convergent inputs. Twelve of 21 neurons received monosynaptic excitatory inputs from both nerves. About half (9/21, 43%) of the convergent neurons were classified as VS neurons, the majority of which descended through the ipsilateral lateral vestibulospinal tract (i-LVST). The HC/SAC convergent neurons were located in the rostral part of the descending, the medial and the caudal-ventral part of the lateral vestibular nucleus. In 80 neurons studied by stimulation of the HC/UT nerves, both inputs converged in 12 (15%) neurons, more than half of which were VS neurons. Eight of 12 convergent neurons received excitatory inputs followed by inhibition from both the HC and UT nerves. A few convergent neurons (3/12) projected to the oculomotor/trochlear nucleus. Half of the convergent and non-convergent VS neurons descended to the spinal cord through the i-LVST, and the only one VOS convergent neuron via the medial vestibulospinal tract. Most of the convergent neurons were located in the lateral, the rostral part of the descending and medial vestibular nucleus. The percentages of HC/SAC and HC/UT convergence were half those of the posterior semicircular canal (PC), PC/SAC (33%) and PC/UT (33%) convergence, respectively. The convergent neurons receiving the HC and otolith inputs may contribute at least partly to the vestibulocollic reflex. PMID- 11500793 TI - Anatomical evidence for brainstem circuits mediating feeding motor programs in the leopard frog, Rana pipiens. AB - Using injections of small molecular weight fluorescein dextran amines, combined with activity-dependent uptake of sulforhodamine 101 (SR101), brainstem circuits presumed to be involved in feeding motor output were investigated. As has been shown previously in other studies, projections to the cerebellar nuclei were identified from the cerebellar cortex, the trigeminal motor nucleus, and the vestibular nuclei. Results presented here suggest an additional pathway from the hypoglossal motor nuclei to the cerebellar nucleus as well as an afferent projection from the peripheral hypoglossal nerve to the Purkinje cell layer of the cerebellar cortex. Injections in the cerebellar cortex combined with retrograde labeling of the peripheral hypoglossal nerve demonstrate anatomical convergence at the level of the medial reticular formation. This suggests a possible integrative region for afferent feedback from the hypoglossal nerve and information through the Purkinje cell layer of the cerebellar cortex. The activity-dependent uptake of SR101 additionally suggests a reciprocal, polysynaptic pathway between this same area of the medial reticular formation and the trigeminal motor nuclei. The trigeminal motor neurons innervate the m adductor mandibulae, the primary mouth-closing muscle. The SR101 uptake clearly labeled the ventrolateral hypoglossal nuclei, the medial reticular formation, and the Purkinje cell layer of the cerebellar cortex. Unlike retrograde labeling of the peripheral hypoglossal nerve, stimulating the hypoglossal nerve while SR101 was bath-applied labeled trigeminal motor neurons. This, combined with the dextran labeling, suggests a reciprocal connection between the trigeminal motor nuclei and the cerebellar nuclei, as well as the medulla. Taken together, these data are important for understanding the neurophysiological pathways used to coordinate the proper timing of an extremely rapid, goal-directed movement and may prove useful for elucidating some of the first principles of sensorimotor integration. PMID- 11500794 TI - Neck muscle responses to abrupt vertical acceleration in the seated human. AB - The control of neck muscle during marked changes in "g" loading must be to protect head-trunk orientation. However, little is known about the organization of reflexes. We therefore investigated the shortest latency in neck muscle evoked by abrupt ascending and descending vertical acceleration with a stroke of 20 cm and peak acceleration 0.4xg in six healthy subjects. The subjects were seated upright and restrained on a chair driven by a hydraulic servo system. Ascent induced small responses in sternocleidomastoid muscles (SCMs) with a latency of 24.0 ms (SE 1.7) from onset of head acceleration, followed by larger responses of 100 ms in duration. In comparison the responses in SCMs evoked by descent were significantly delayed, with greater interindividual variations at 44.1 ms (SE 3.1) and smaller in amplitude than responses in ascent. Latencies were consistent and showed no habituation. In order to eliminate stretch components, we measured the neck responses to vertical acceleration with the head fixed by cervical collar. The latency of the fast response of SCM evoked by ascent was not significantly different from the latency of SCMs without the collar. These results may indicate that the fast responses of SCMs to sudden ascent may be composed of a vestibular-collic reflex for making the neck and head rigid for defense to sudden gravitational change. PMID- 11500795 TI - Head unrestrained horizontal gaze shifts after unilateral labyrinthectomy in the rhesus monkey. AB - Following the orienting saccade of a combined eye-head gaze shift, normal monkeys exhibit a compensatory eye counterrotation that stabilizes gaze as the head movement continues. This counterrotation, which has a gain (eye velocity/head velocity) of near unity, is a manifestation of the vestibulo-ocular reflex (VOR). Acute unilateral labyrinthectomy (UL) causes severe asymmetry in the VOR during passive head rotations that recovers incompletely over time. The purpose of this investigation was to compare the recovery of the counterrotation gain during horizontal gaze shifts with that of the passive VOR after UL. During the 1st week after UL, counterrotation gains were asymmetric, being lower for head movements towards the lesion but nearly normal for head movements towards the intact side. Whereas this asymmetry in the counterrotation gain resolved within a week after UL, asymmetries in the passive VOR persisted. During the 1st week after UL, behavioral performance was generally poor, with a high incidence of inaccurate gaze shifts and larger latencies. In addition, animals used slower head movements such that peak head amplitude during the eye saccade was significantly lower during the 1st week after UL as compared to control values. Bilateral labyrinthectomy (BL) resulted in larger but symmetric deficits in counterrotation, which, contrary to the passive VOR, exhibited significant recovery over time. It is hypothesized that recovery of counterrotation gain after UL has contributions from multiple sources, including the contralateral intact labyrinth and an efference copy of the head movement. PMID- 11500796 TI - Effect of single-limb inertial loading on bilateral reaching: interlimb interactions. AB - This study employed the paradigm of asymmetric limb loading during bilateral arm reaching to examine the motor system's ability to independently organize the discrete movement of both upper limbs to equidistant targets when one of the limbs is loaded under specific timing constraints. The loading procedure involved attaching two different Velcro strapped weights to the right wrist, thus increasing the right arm's mass by 25% (1 kg) and 50% (2 kg). Movements were captured by a high-speed digital camera (240 Hz), while electromyographic (EMG) activity of selected elbow and shoulder muscles of both limbs was recorded (1,000 Hz) simultaneously. The results revealed that the mechanisms used by the system to compensate for unilateral limb loading were as follows: First, addition of an inertial load resulted in an increased movement time and concomitant decrease in peak velocity of both the upper arm and forearm of only the loaded limb and was scaled to the added weight. Second, for the EMG parameters, adjustments to the inertial load were primarily characterized by an increase in burst duration of all muscles, with load-specific changes in activity and onset time: the elbow antagonist (biceps) demonstrated a decrease in activity with the 50% load, and the elbow agonist (triceps) had an earlier onset with the 25% load. Concomitant adjustments on the unloaded limb consisted primarily of an increase in burst duration of the shoulder and elbow agonists (pectoralis and triceps), an earlier triceps onset solely with the 25% load, and a decrease in activity of the biceps solely with the 50% load. Third, with the exception of biceps activity, the amplitude of EMG activity was invariant across changes in load for both the loaded and unloaded limb. This lack of modulation in activity may have been related to the inability of performers to meet the time constraint of simultaneous bilateral limb arrival to the end targets. This inability can be the result of an active strategy selection process to safeguard the actions against interference or alternatively it could simply be a consequence of the biomechanical properties of the system in relation to task constraints. These issues are discussed in the light of the present findings and those of previous studies. PMID- 11500797 TI - Local field potentials related to bimanual movements in the primary and supplementary motor cortices. AB - We recorded local field potentials (LFP) in primary (MI) and supplementary (SMA) motor areas of rhesus monkey cortex in order to compare movement-evoked potentials (mEP) in bimanual and unimanual movements with single-unit activity recorded concurrently. The mEP was often different during bimanual and unimanual movements (a "bimanual-related" effect), but, unlike the single units, the size of the mEP in both MI and SMA was always greater during bimanual movements than during unimanual movements. This increase primarily reflected an increase in the late positive peak of the mEP, a result that may reflect greater overall cortical activation during bimanual movements. In addition, analysis of the mEP revealed differences between MI and SMA not seen in the single-unit activity. mEP in MI had greater contralateral preference than in SMA. Also, SMA mEP was more correlated to the single-unit activity than in MI. This greater correlation was also more apparent in the late peaks of the mEP than in the early peaks and may reflect a greater influence of recurrent activation in SMA than in MI. Our results further reinforce the idea that unimanual and bimanual movements are represented differently both in MI and in SMA and also show that a complex relationship between spikes of individual neurons and LFP may reflect the different input-output relations of different cortical areas. PMID- 11500798 TI - Scalp potentials to pitch change in rapid tone sequences. A correlate of sequential stream segregation. AB - The object of the study was to look for a neurophysiological substrate of sequential auditory stream segregation. When a sequence of tones alternates rapidly between pitches separated by more than a few semitones, there is a tendency for it to be perceived as two independent "streams". We examined the scalp potentials evoked when the pitch interval abruptly changes, to see whether there are response parameters which might be correlated with sudden stream segregation and/or integration. For 3 s a continuous synthesized tone of "clarinet" timbre oscillated between pitches of F4 and F#4 (one semitone higher) at 16 notes/s, perceived as an integrated stream. The upper note was then raised to E5 (11 semitones above F4, perceived as segregated streams) for a further 3 s and the cycle was repeated 40 times. In a second condition also starting with oscillation between F4 and F#4, the upper note was lowered to E4 (one semitone below F4, still perceived as a single stream). Further conditions examined the changes between oscillations of 1 and 11 semitones down from E5, 1 and 23 semitones up from F4, and 10 and 11 semitones up from F4. Virtually no potentials were detectable during the periods of unchanging oscillation, but an N1/P2 complex was evoked on each change in the pitch interval. The N1 was termed "MN1" on account of its arguable relatedness to the mismatch negativity, recorded in a separate experiment using discontinuous tones at a much slower rate. The mean peak latency of the MN1 varied between 96 and 123 ms, the shortest latencies being recorded, not to the largest changes of pitch interval but to the widest pitch intervals between the new tone and the immediately preceding one. Therefore, although a causal relationship with streaming cannot necessarily be inferred, the MN1 latency appears to mark the degree of pitch contrast between consecutive tones, in correlation with the streaming effect. PMID- 11500799 TI - Long-term adaptation to dynamics of reaching movements: a PET study. AB - Positron emission tomography (PET) was used to examine changes in the cerebellum as subjects learned to make movements with their right arm while holding the handle of a robot that produced a force field. Brain images were acquired during learning and then during recall at 2 and 4 weeks. We also acquired images during a control task where the force field was not learnable and subjects did not show any improvements across sessions. During the 1st day, we observed that motor errors decreased from the control condition to the learning condition. However, regional cerebral blood flow (rCBF) in the posterior region of the right cerebellar cortex initially increased from the control condition and then gradually declined with reductions in motor error. Correspondingly, rCBF in the ipsilateral deep cerebellar nuclei (DCN) initially decreased from the control condition and then increased with reductions in motor error. If measures of rCBF mainly reflect presynaptic activity of neurons, this result predicts that DCN neurons fire with a pattern that starts high in the control task then decreases as learning proceeds. Similarly, Purkinje cells should generally have their lowest activity in the control task. This pattern is consistent with neurophysiological recordings that find that cerebellar activity during early learning of a motor task may mainly reflect changes in coactivation of muscles of the limbs, rather than a learning specific signal. By the end of the first session, motor errors had reached a minimum and no further improvements were observed. However, across the weeks a region in the anterior cerebellar cortex showed gradual decreases in rCBF that could not be attributed to changes in motor performance. Because patterns of rCBF in the cortex and nuclei were highly anti correlated, we used structural equation modeling to estimate how synaptic activity in the cerebellar cortex influenced synaptic activity in the DCN. We found a negative correlation with a strength that significantly increased during the 4 weeks. This suggests that, during long-term recall, the same input to the cerebellar cortex would produce less synaptic activity at the DCN, possibly because of reduced cerebellar cortex output to the DCN. PMID- 11500800 TI - Syntactic and semantic processing in the healthy and aphasic human brain. AB - A syntactic and a semantic task were performed by German-speaking healthy subjects and aphasics with lesions in the dominant left hemisphere. In both tasks, pictures of objects were presented that had to be classified by pressing buttons. The classification was into grammatical gender in the syntactic task (masculine or feminine gender?) and into semantic category in the semantic task (man- or nature made?). Behavioral data revealed a significant Group by Task interaction, with aphasics showing most pronounced problems with syntax. Brain event-related potentials 300-600 ms following picture onset showed different task dependent laterality patterns in the two groups. In controls, the syntax task induced a left-lateralized negative ERP, whereas the semantic task produced more symmetric responses over the hemispheres. The opposite was the case in the patients, where, paradoxically, stronger laterality of physiological brain responses emerged in the semantic task than in the syntactic task. We interpret these data based on neuro-psycholinguistic models of word processing and current theories about the roles of the hemispheres in language recovery. PMID- 11500801 TI - Bilateral multifinger deficits in symmetric key-pressing tasks. AB - Maximal voluntary force during simultaneous bilateral and multifinger exertion has been shown to be smaller than the sum of unilateral or single-finger exertions. The goal of this study was to study the force deficit associated with bilateral multifinger tasks. Eight normal college students performed four types of maximal isometric key-pressing tasks: (1) unilateral single-finger, (2) bilateral single-finger, (3) unilateral multifinger, and (4) bilateral multifinger. Forces produced by the index (I), middle (M), ring (R), and little (L) fingers and surface electromyography (EMG) of extrinsic finger flexors were recorded. Multifinger deficit (MFD) was defined as the percentage difference between the force (or EMG) produced by a set of fingers and the sum of the forces (or EMGs) produced by the individual fingers in their unilateral single-finger tasks. Bilateral deficit (BLD) was defined as the percentage difference between the force (or EMG) produced by a set of fingers and the sum of the forces (or EMGs) produced by the finger subsets of the left and right hands. Significant BLD and MFD in force and EMG were found for all bilateral multifinger tasks and some of the bilateral single-finger tasks. Both BLD and MFD were dependent on the number of fingers involved. BLD ranged from 3% to 22.7% for force and from 8.9% to 31.0% for EMG, including bilateral single-finger and bilateral multifinger tasks. MFDs in force during bilateral I-, IM-, IMR-, and IMRL-finger tasks were 13.2%, 37.8%, 53.2%, 52.3%, respectively; and the corresponding MFDs in EMG were 11.7%, 51.3%, 67.6%, and 71.0%, respectively. BLD and MFD in EMG were found to vary in parallel with the corresponding force deficits. It was suggested that the neural ceiling effect remains the most plausible mechanism underlying the observed deficits. The central nervous system is unable to activate maximally a large number of muscle groups at the same time during tasks involving multiple body parts. During bilateral multifinger tasks, the ceiling effect may be organized hierarchically: (1) a certain limited neural drive is shared bilaterally, leading to a BLD; (2) at each hand, a certain limited neural drive is shared by multiple fingers, leading to MFD within a hand; (3) the deficits at bilateral and unilateral multifinger levels are cumulative during bilateral multifinger tasks, leading to a higher deficit associated with the tasks. PMID- 11500802 TI - Vestibular influences on human postural control in combinations of pitch and roll planes reveal differences in spatiotemporal processing. AB - The present study examined the influence of bilateral peripheral vestibular loss (BVL) in humans on postural responses to multidirectional surface rotations in the pitch and roll planes. Specifically, we examined the effects of vestibular loss on the directional sensitivity, timing, and amplitude of early stretch, balance correcting, and stabilizing reactions in postural leg and trunk muscles as well as changes in ankle torque and trunk angular velocity following multidirectional rotational perturbations of the support surface. Fourteen normal healthy adults and five BVL patients stood on a dual axis rotating platform which rotated 7.5 degrees at 50 degrees/s through eight different directions of pitch and roll combinations separated by 45 degrees. Directions were randomized within a series of 44 perturbation trials which were presented first with eyes open, followed by a second series of trials with eyes closed. Vestibular loss did not influence the range of activation or direction of maximum sensitivity for balance correcting responses (120-220 ms). Response onsets at approximately 120 ms were normal in tibialis anterior (TA), soleus (SOL), paraspinals (PARAS), or quadriceps muscles. Only SOL muscle activity demonstrated a 38- to 45-ms delay for combinations of forward (toe-down) and roll perturbations in BVL patients. The amplitude of balance correcting responses in leg muscles between 120 and 220 ms was, with one exception, severely reduced in BVL patients for eyes open and eyes closed conditions. SOL responses were decreased bilaterally for toe-up and toe-down perturbations, but more significantly reduced in the downhill (load bearing) leg for combined roll and pitch perturbations. TA was significantly reduced bilaterally for toe-up perturbations, and in the downhill leg for backward roll perturbations. Forward perturbations, however, elicited significantly larger TA activity in BVL between 120 and 220 ms compared to normals, which would act to further destabilize the body. As a result of these changes in response amplitudes, BVL patients had reduced balance correcting ankle torque between 160 and 260 ms and increased torque between 280 and 380 ms compared to normals. There were no differences in the orientation of the resultant ankle torque vectors between BVL and normals, both of which were oriented primarily along the pitch plane. For combinations of backward (toe-up) and roll perturbations BVL patients had larger balance correcting and stabilizing reactions (between 350 and 700 ms) in PARAS than normals and these corresponded to excessive trunk pitch and roll velocities. During roll perturbations, trunk velocities in BVL subjects after 200 ms were directed along directions different from those of normals. Furthermore, roll instabilities appeared later than those of pitch particularly for backward roll perturbations. The results of the study show that combinations of roll and pitch surface rotations yield important spatiotemporal information, especially with respect to trunk response strategies changed by BVL which are not revealed by pitch plane perturbations alone. Our results indicate that vestibular influences are earlier for the pitch plane and are directed to leg muscles, whereas roll control is later and focused on trunk muscles. PMID- 11500803 TI - Transcranial magnetic stimulation in the rat. AB - Transcranial magnetic stimulation (TMS) allows for quantification of motor system excitability. While routinely used in humans, application in other species is rare and little is known about the characteristics of animal TMS. The unique features of TMS, i.e., predominantly interneuronal stimulation at low intensity and non-invasiveness, are particularly useful in evaluating injury and recovery in animal models. This study was conducted to characterize the rodent motor evoked potential to TMS (MEPTMS) and to develop a methodology for reproducible assessment of motor excitability in the rat. MEPTMS were compared with responses evoked by electrical stimulation of cervical spinal cord (MEPCES) and peripheral nerve. MEP were recorded by subcutaneous electrodes implanted bilaterally over the calf. Animals remained under propofol infusion and restrained in a stereotactic frame while TMS followed by CES measurements were obtained before and after 2 h of idle time. TMS was applied using a 5-cm-diameter figure-of-eight coil. MEPTMS had onset latencies of 6.7+/-1.3 ms. Latencies decreased with higher stimulation intensity (r=-0.7, P<0.05). Two morphologies, MEPTMS, 1 and MEPTMS, 2, were distinguished by latency of the first negative peak (N1), overall shape, and amplitude. MEPTMS, 2 were more frequent at higher stimulation intensity. While recruitment curves for MEPTMS, 1 followed a sigmoid course, no supramaximal response was reached for MEPTMS, 2. Mid-cervical spinal transection completely abolished any response to TMS. MEPCES showed a significantly shorter latency (5.29+/-0.24, P<0.0001). Two types of MEPCES resembling MEPTMS, 1 and 2 were observed. Neither MEPTMS nor MEPCES changed on repeat assessment after 2 h. This study demonstrates the feasibility and reproducibility of TMS in the rat. Sigmoid recruitment curves for MEPTMS, 1 suggest input-output properties similar to those of the human corticospinal system. Latency differences between CES and TMS point to a supraspinal origin of the MEPTMS. The two morphologies likely reflect different cortical or subcortical origins of MEPTMS. PMID- 11500804 TI - Nociceptive visceral stimulation modulates the activity of cerebellar Purkinje cells. AB - The cerebellum is a system with various input and output functions that influence motor, sensory, cognitive, and other processes. In a previous study, we showed that cerebellar cortical stimulation increases spinal neuronal responses to visceral noxious stimulation by colorectal distension (CRD). However, the neuronal network underlying the cerebellar modulation of nociceptive phenomena is largely unknown. Purkinje cells of the cerebellar cortex receive ascending and descending inputs and exert a major inhibitory control over neurons in the underlying cerebellar nuclei that constitute the cerebellar output. Therefore, in this study, we tested the effect of CRD and other somatic stimuli on the firing rate of Purkinje cells using in vivo extracellular recording techniques. The results suggest that Purkinje cells respond to nociceptive visceral and somatic stimulation in the form of early and delayed changes in activity. Based on these and previous findings, we propose a negative feedback circuitry involving the cerebellum for the modulation of peripheral nociceptive events. PMID- 11500805 TI - MC1R genotype modifies risk of melanoma in families segregating CDKN2A mutations. AB - Mutations in the exons of the cyclin-dependent kinase inhibitor gene CDKN2A are melanoma-predisposition alleles which have high penetrance, although they have low population frequencies. In contrast, variants of the melanocortin-1 receptor gene, MC1R, confer much lower melanoma risk but are common in European populations. Fifteen Australian CDKN2A mutation-carrying melanoma pedigrees were assessed for MC1R genotype, to test for possible modifier effects on melanoma risk. A CDKN2A mutation in the presence of a homozygous consensus MC1R genotype had a raw penetrance of 50%, with a mean age at onset of 58.1 years. When an MC1R variant allele was also present, the raw penetrance of the CDKN2A mutation increased to 84%, with a mean age at onset of 37.8 years (P=.01). The presence of a CDKN2A mutation gave a hazard ratio of 13.35, and the hazard ratio of 3.72 for MC1R variant alleles was also significant. The impact of MC1R variants on risk of melanoma was mediated largely through the action of three common alleles, Arg151Cys, Arg160Trp, and Asp294His, that have previously been associated with red hair, fair skin, and skin sensitivity to ultraviolet light. PMID- 11500806 TI - Melanocortin-1 receptor variant R151C modifies melanoma risk in Dutch families with melanoma. AB - Germline mutations of the cell-cycle regulator p16 (also called "CDKN2A") in kindreds with melanoma implicate this gene in susceptibility to malignant melanoma. Most families with familial atypical multiple-mole melanoma (FAMMM) who are registered at the Leiden dermatology clinic share the same p16-inactivating deletion (p16-Leiden). Incomplete penetrance and variable clinical expression suggest risk modification by other genetic and/or environmental factors. Variants of the melanocortin-1 receptor (MC1R) gene have been shown to be associated with red hair, fair skin, and melanoma in humans. Carriers of the p16-Leiden deletion in Dutch families with FAMMM show an increased risk of melanoma when they also carry MC1R variant alleles. The R151C variant is overrepresented in patients with melanoma who are from families with the p16-Leiden mutation. Although some of the effect of the R151C variant on melanoma risk may be attributable to its effect on skin type, our analyses indicate that the R151C variant contributes an increased melanoma risk even after statistical correction for its effect on skin type. These findings suggest that the R151C variant may be involved in melanoma tumorigenesis in a dual manner, both as a determinant of fair skin and as a component in an independent additional pathway. PMID- 11500807 TI - The amyloid precursor protein locus and very-late-onset Alzheimer disease. AB - Although mutations in the amyloid-beta precursor protein (APP) gene are known to confer high risk of Alzheimer disease (AD) to a small percentage of families in which it has early onset, convincing evidence of a major role for the APP locus in late-onset AD has not been forthcoming. In this report, we have used a covariate-based affected-sib-pair linkage method to analyze the chromosome 21 clinical and genetic data obtained on affected sibships by the National Institute of Mental Health Alzheimer Disease Genetics Initiative. The baseline model (without covariates) gave a LOD score of 0.02, which increases to 1.43 when covariates representing the additive effects of E2 and E4 are added. Larger increases in LOD scores were found when age at last examination/death (LOD score 5.54; P=.000002) or age at onset plus disease duration (LOD score 5.63; P=.000006) were included in the linkage model. We conclude that the APP locus may predispose to AD in the very elderly. PMID- 11500808 TI - Anatomy of tissue engineering. PMID- 11500809 TI - Regaining chondrocyte phenotype in thermosensitive gel culture. AB - Chondrocyte tissue engineering continues to be a challenging problem. When chondrocytes are duplicated in vitro, it is imperative to obtain an adequate number of cells of optimal phenotype. A temperature-sensitive polymer gel, a copolymer of poly(N-isopropylacrylamide) and acrylic acid (PNiPAAm-co-Aac), has the ability of gelling at 37 degrees C (the lower critical solution temperature, LCST) or above and liquefying below that temperature (Vernon and Gutowska, Macromol. Symp. 1996;109:155-167). The hypothesis of this study was that chondrocytes could (1) duplicate in the copolymer gel; (2) regain their chondrocyte phenotype; and (3) be easily recovered from the gel by simply lowering the temperature below 37 degrees C. Chondrocytes from adult rabbit scapular cartilage were harvested and cultured in a monolayer culture until confluency (approximately 2 weeks). Next, the cells were harvested and seeded into the copolymer gel and cultured for 2-4 weeks. The phenotype of the cultured cells was then characterized. Two groups of control cultures, monolayer and agarose gel, were used to compare their ability to maintain chondrocyte phenotype. The results showed that chondrocytes isolated from rabbit scapula can re-express chondrocyte phenotype in agarose culture and polymer gel culture but not in monolayer culture. Also, cultured chondrocytes can be easily recovered from polymer gel culture by simply lowering the temperature. This new in vitro method of chondrocyte culture is recommended for chondrocyte propagation and regaining chondrocyte phenotype before cell seeding or transplantation. PMID- 11500810 TI - Injectable gels for tissue engineering. AB - Recently, tissue engineering approaches using injectable, in situ gel forming systems have been reported. In this review, the gelation processes and several injectable systems that exhibit in situ gel formation at physiological conditions are discussed. Applications of selected injectable systems (alginate, chitosan, hyaluronan, polyethylene oxide/polypropylene oxide) in tissue engineering are also described. Injectable polymer formulation can gel in vivo in response to temperature change (thermal gelation), pH change, ionic cross-linking, or solvent exchange. Kinetics of gelation is directly affected by its mechanism. Injectable formulations offer specific advantages over preformed scaffolds such as: possibility of a minimally invasive implantation, an ability to fill a desired shape, and easy incorporation of various therapeutic agents. Several factors need to be considered before an injectable gel can be selected as a candidate for tissue engineering applications. Apart from tissue-specific cell-matrix interactions, the following gel properties need to be considered: gelation kinetics, matrix resorption rate, possible toxicity of degradation products and their elimination routes, and finally possible interference of the gel matrix with histogenesis. PMID- 11500811 TI - Clonogenic analysis reveals reserve stem cells in postnatal mammals: I. Pluripotent mesenchymal stem cells. AB - Clonal populations of lineage-uncommitted pluripotent mesenchymal stem cells have been identified in prenatal avians and rodents. These cells reside in the connective tissue matrices of many organs and tissues. They demonstrate extended capabilities for self-renewal and the ability to differentiate into multiple separate tissues within the mesodermal germ line. This study was designed to determine whether such cells are present in the connective tissues of postnatal mammals. This report describes a cell clone derived by isolation from postnatal rat connective tissues, cryopreservation, extended propagation, and serial dilution clonogenic analysis. In the undifferentiated state, this clone demonstrates a high nuclear-to-cytoplasmic ratio and extended capacity for self renewal. Subsequent morphological, histochemical, and immunochemical analysis after the induction of differentiation revealed phenotypic markers characteristic of multiple cell types of mesodermal origin, such as skeletal muscle, smooth muscle, fat cells, cartilage, and bone. These results indicate that this clone consists of pluripotent mesenchymal stem cells. This report demonstrates that clonal populations of reserve stem cells are present in mammals after birth. Potential roles for such cells in the maintenance, repair, and regeneration of mesodermal tissues are discussed. PMID- 11500812 TI - Tissue engineering strategies for adipose tissue repair. AB - Tissue engineering is a relatively young field that combines engineering, clinical science, and life sciences to, in part, repair or regrow tissues. Adipose tissue has recently become a focus area for tissue engineering, encouraged by the large number of reconstructive, cosmetic, and correctional indications that could be addressed with clinically translatable adipose tissue engineering strategies. This review discusses the three aspects of an adipose construct, namely cell types, scaffold, and microenvironment, and presents current tissue engineering strategies under pursuit. PMID- 11500813 TI - Tissue engineering in the cardiovascular system: progress toward a tissue engineered heart. AB - Achieving the lofty goal of developing a tissue engineered heart will likely rely on progress in engineering the various components: blood vessels, heart valves, and cardiac muscle. Advances in tissue engineered vascular grafts have shown the most progress to date. Research in tissue-engineered vascular grafts has focused on improving scaffold design, including mechanical properties and bioactivity; genetically engineering cells to improve graft performance; and optimizing tissue formation through in vitro mechanical conditioning. Some of these same approaches have been used in developing tissue engineering heart valves and cardiac muscle as well. Continued advances in scaffold technology and a greater understanding of vascular cell biology along with collaboration among engineers, scientists, and physicians will lead to further progress in the field of cardiovascular tissue engineering and ultimately the development of a tissue-engineered heart. PMID- 11500814 TI - Tissue engineering in plastic reconstructive surgery. AB - Tissue engineering (TE) is a new interdisciplinary field of applied research combining engineering and biosciences together with clinical application, mainly in surgical specialities, to develop living substitutes for tissues and organs. Tissue engineering approaches can be categorized into substitutive approaches, where the aim is the ex vivo construction of a living tissue or organ similar to a transplant, vs. histioconductive or histioinductive concepts in vivo. The main successful approaches in developing tissue substitutes to date have been progresses in the understanding of cell-cell interactions, the selection of appropriate matrices (cell-matrix interaction) and chemical signalling (cytokines, growth factors) for stimulation of cell proliferation and migration within a tissue-engineered construct. So far virtually all mammalian cells can be cultured under specific culture conditions and in tissue specific matrices. Future progress in cell biology may permit the use of pluripotent stem cells for TE. The blueprint for tissue differentiation is the genome: for this it is reasonable to combine tissue engineering with gene therapy. The key to the progress of tissue engineering is an understanding between basic scientists, biochemical engineers, clinicians, and industry. PMID- 11500815 TI - Engineering of vascular ingrowth matrices: are protein domains an alternative to peptides? AB - Anastomotic intimal hyperplasia and surface thrombogenicity are the main reasons for the high failure rate of prosthetic small-diameter vascular grafts. While anastomotic intimal hyperplasia is a multifactorial event, ongoing surface thrombogenicity is primarily caused by the lack of an endothelium, even after years of clinical implantation. After decades of poorly performing synthetic artery-grafts, tissue engineering has emerged as a promising approach to generate biologically functional bio-synthetic hybrid grafts mimicking native arteries regarding the presence of an endothelial lining on the blood surface. "In vitro endothelialization" represented the first generation of such tissue-engineered vascular grafts, utilising cell culture techniques for the creation of a confluent autologous endothelium on ePTFE grafts. The clinical long-term results with this method in almost 200 patients are highly encouraging, showing patencies equal to vein grafts. Since "in vitro endothelialization" requires cell culture facilities, it will always be confined to large centres. Therefore, research of the 1990s turned to the development of spontaneously endothelializing implants, to make tissue-engineered grafts amenable to the entire vascular-surgical community. Apart from scaffold designs allowing transmural ingrowth, biological signalling through a facilitating ingrowth matrix holds a key to spontaneous endothelialization. In biological signalling, the increasingly deeper understanding of bio-active molecules and the discovery of domains and peptide sequences during the 1980s created the expectation in the 1990s that peptide signalling may be all that is needed. This present review highlights the possible problems associated with such a reductionist approach. Using the fibronectin molecule, we demonstrated that domains may be more suitable modules in tissue engineering than peptide sequences. PMID- 11500816 TI - Spinal fusion with recombinant human growth and differentiation factor-5 combined with a mineralized collagen matrix. AB - The availability of recombinant osteoinductive growth factors and new osteoconductive matrices offers an alternative to the use of autogenous bone (autograft) for grafting indications. This study evaluates the bone-forming activity of a mineralized collagen matrix combined with recombinant human growth and differentiation factor-5 in a rabbit posterolateral spinal fusion model. The activity of three distinct matrix-growth factor formulations is assessed by radiographic, histologic, and mechanical strength methods. Results show that the radiographic density, histologic quality, and mechanical strength of fusion at 12 weeks post-treatment rank consistently within the treatment groups. Optimal formulations are shown to perform similar to autograft in both the rate and strength of fusion. Fusion rates as high as 80% are observed within specific matrix/growth factor formulations. The average biomechanical strength of treated motion segments in the most efficacious formulation is 82% higher than that obtained with autograft, although this difference is not statistically significant. The fusion mass formed in response to matrix/growth factor formulations is composed of normal trabecular bone with a thin outer cortical plate and modest hematopoietic bone marrow. These results demonstrate that the combination of a mineralized collagen matrix with recombinant human growth and differentiation factor-5 maximizes the inherent conductive and inductive properties of each component, respectively, to provide an effective alternative to autograft for bone grafting procedures. PMID- 11500817 TI - Peripheral nerve injury: a review and approach to tissue engineered constructs. AB - Eleven thousand Americans each year are affected by paralysis, a devastating injury that possesses associated annual costs of $7 billion (American Paralysis Association, 1997). Currently, there is no effective treatment for damage to the central nervous system (CNS), and acute spinal cord injury has been extraordinarily resistant to treatment. Compared to spinal cord injury, damage to peripheral nerves is considerably more common. In 1995, there were in excess of 50,000 peripheral nerve repair procedures performed. (National Center for Health Statistics based on Classification of Diseases, 9th Revision, Clinical Modification for the following categories: ICD-9 CM Code: 04.3, 04.5, 04.6, 04.7). These data, however, probably underestimate the number of nerve injuries appreciated, as not all surgical or traumatic lesions can be repaired. Further, intraabodominal procedures may add to the number of neurologic injuries by damage to the autonomic system through tumor resection. For example, studies assessing the outcome of impotency following radical prostatectomy demonstrated 212 of 503 previously potent men (42%) suffered impotency when partial or complete resection of one or both cavernosal nerve(s). This impotency rate decreased to 24% when the nerves were left intact (Quinlan et al., J. Urol. 1991;145:380-383; J. Urol. 1991;145:998-1002). PMID- 11500818 TI - A gender--dependent genetic predisposition to produce high levels of IL-6 is detrimental for longevity. AB - Current literature indicates that elevated IL-6 serum levels are associated with diseases, disability and mortality in the elderly. In this paper, we studied the IL-6 promoter genetic variability at -174 C/G locus and its effect on IL-6 serum levels in a total of 700 people from 60 to 110 years of age, including 323 centenarians. We found that the proportion of homozygotes for the G allele at 174 locus decreases in centenarian males, but not in centenarian females. Moreover, we found that, only among males, homozygotes for the G allele at -174 locus have higher IL-6 serum levels in comparison with carriers of the C allele. On the whole, our data suggest that those individuals who are genetically predisposed to produce high levels of IL-6 during aging, i.e. -174 locus GG homozygous men, are disadvantaged for longevity. PMID- 11500819 TI - Emergence of a type II collagen-specific helper T cell response. AB - Antigen-driven development of persistent self-reactive Th cells underlies the chronic, progressive nature of many autoimmune diseases. It is crucial to understand the behavior of these self-reactive Th cells; however, they have been notoriously difficult to isolate directly ex vivo. Collagen-induced arthritis (CIA) can be initiated in I-A(q)-expressing mice (DBA/1) using heterologous type II collagen (cII) immunization and is dependent on Th cells that are specific for a single immunodominant epitope. Here, we identify one compartment of cII specific Th cell using TCR beta expression, cell surface phenotype, and direct single-cell repertoire analysis. A subpopulation of CD4(+)V beta 10(+) T cells up regulates both CD44 and GL7 and expands significantly in response to initial priming in the majority of animals (D9: 70%). The cII-specific V beta 10(+) primary responders are further resolved through expression of a highly restricted junctional region, previously associated with autoimmune disease. This cII specific clonotype rapidly re-expands upon antigen recall and can be isolated from the lymph nodes of arthritic animals. These single-cell analyses quantify the emergence, decline and rapid re-emergence of a self-reactive Th cell population in vivo and outline one strategy for isolating these cells directly ex vivo. PMID- 11500820 TI - Carrier-mediated enhancement of cognate T cell help: the basis for enhanced immunogenicity of meningococcal outer membrane protein polysaccharide conjugate vaccine. AB - Haemophilus influenzae type b capsular polysaccharide (PRP) conjugate vaccines, which are thought to induce T cell-dependent antibody production, induce protective responses after a single dose in individuals under 15 months of age. However, multiple doses of these vaccines are required to induce protective antibody responses in infants, with the exception of PRP conjugated to meningococcal outer membrane proteins (OMPC), which does so after a single dose. The basis for this difference is not fully understood, although others have proposed that OMPC and porins, the major protein component of OMPC, act as adjuvants or mitogens. In this report OMPC is shown to enhance CD40 ligand mediated, T cell-dependent antibody production in mice. This paralleled the induction by OMPC of CD86, CD80 and CD40 costimulatory molecules on human neonatal and murine B cells and of Th1 cytokines. Neither porins nor lipopolysaccharide fully reproduced the effects of OMPC. These studies indicate that OMPC acts both as carrier and adjuvant, and thereby enhances T cell dependent antibody responses in human infants. PMID- 11500821 TI - Hck SH3 domain-dependent abrogation of Nef-induced class 1 MHC down-regulation. AB - The ability of specific virally encoded proteins to down-regulate MHC class I molecules may enable infected cells to elude killing by CTL. In the case of HIV 1, Nef appears to be responsible for this effect. Thus, interfering with Nef induced MHC class I down-regulation would be a strategy for increasing HIV-1 specific CTL activity, particularly towards long-lived T cell populations such as memory T cells that harbor replication-competent virus. Here, using two Nef expressing human cell model systems, we show that a dominant-negative mutant derived from the Hck protein-tyrosine kinase, composed of the Hck N-terminal region, as well as the SH3 and SH2 domains, was able to inhibit Nef-induced MHC class I molecule down-regulation. This effect was SH3 domain dependent as it was not evident when the cells were transfected with DN-Hck-W93F, an SH3 domain mutant. The inhibitory effect of dominant-negative-Hck (DN-Hck) on Nef-induced class I down-regulation suggests that this Nef-mediated effect requires an interaction between the Nef polyproline site and an SH3-containing cellular protein that is involved in MHC class I molecule turnover. Interfering with the function of the Nef SH3 binding site in this way represents a strategy for assisting the host CTL response to clear HIV-1-infected cells. PMID- 11500822 TI - Direct ex vivo comparison of the breadth and specificity of the T cells in the liver and peripheral blood of patients with chronic HCV infection. AB - The role of intrahepatic lymphocytes in the control of hepatitis C virus (HCV) infection and the pathology associated with it is not understood; most studies of the immunology of this infection use peripheral blood lymphocyte populations. To address this further, we examined in detail the IHL from HCV-infected patients and controls, focusing on the antigen-specific CD8(+) T lymphocyte component. Individual T cells from needle liver biopsies and peripheral blood were isolated from patients with chronic HCV infection and examined directly ex vivo. We used RT-PCR spectratyping to compare the breadth of the T cell receptor usage in the liver in comparison with the peripheral blood, and applied MHC class I tetramer technology to investigate the numbers of HCV-specific CD8(+) cells in the two compartments. T cell receptor usage in the liver of HCV-infected patients was broad, comparable with that in the peripheral blood of the same patients. A much higher proportion of liver CD8(+) cells expressed receptors specific for HCV antigens compared with paired peripheral blood CD8(+) cells. A greater proportion of the liver tetramer-positive cells expressed the activation marker CD69, compared with those in the periphery or other CD8(+) cells in the liver. In the course of chronic HCV infection, HCV-specific CD8 cells, which have been recently activated, appear to accumulate specifically in the livers of infected patients but are present in much lower numbers in the peripheral circulation. Further studies are needed to determine the function of these cells and their role in protection and immunopathology. PMID- 11500823 TI - Clonal expansion of freshly isolated CD4T cells by randomized peptides and identification of peptide ligands using combinatorial peptide libraries. AB - We synthesized Xn (n = 9 -- 19) peptides that consist of 9 to 19 residues with random sequences. X19 is considered to deliver antigenic stimuli to CD4 T cells, because: (a) X19 induces proliferation of peripheral blood mononuclear cells (PBMC), in the presence of IL-2, which is abrogated by monoclonal antibodies to class II HLA; (b) X19 + IL-2 induces proliferation of CD4 T cell clones of distinct specificities; and (c) T cell clones recognizing the same TCR ligands with distinct V beta usage are equally stimulated by X19 + IL-2. We next co cultured single peripheral CD4 T cells with X19 and mitomycin-treated autologous PBMC. Indeed, single T cells of CD45RA(-) memory phenotype exhibited clonal expansion, with variable rates of proliferation, when IL-4, IL-7, IL-9, IL-15 and agonistic antibody to CD29 were included in the culture. These T cell clones showed heterogeneous proliferation patterns against KGXXXXXXXXXGK-based and KGXXXXXXXXXGKGKK-based combinatorial peptides libraries, in the presence of IL-2. Pattern-match search on a T cell clone resulted in peptide ligand candidates, one of which induced proliferation, as did protein molecules carrying the corresponding sequence. These results indicate that X19 can induce proliferation of peripheral memory T cells, the peptide ligands of which can be determined using combinatorial peptide libraries. PMID- 11500824 TI - Vav-1 regulates NK T cell development and NK cell cytotoxicity. AB - The hematopoietic-specific Rho-family GTP exchange factor Vav-1 is a regulator of lymphocyte antigen receptor signaling and mediates normal maturation and activation of B and T cells. Recent findings suggest that Vav-1 also forms part of signaling pathways required for natural and antibody dependent cellular cytotoxicity (ADCC) of human NK cells. In this study, we show that Vav-1 is also expressed in murine NK cells. Vav-1(-/-) mice had normal numbers of splenic NK cells, and these displayed a similar expression profile of NK cell receptors as wild-type mice. Unexpectedly, IL-2-activated Vav-1(-/-) NK cells retained normal ADCC. Fc-receptor mediated activation of ERK, JNK, and p38 was also normal. In contrast, Vav-1(-/-) NK cells exhibited reduced natural cytotoxicity against EL4, C4.4.25, RMA and RMA/S. Together, the results demonstrate that Vav-1 is dispensable for mainstream NK cell development, but is required for NK natural cytotoxicity. Unlike the findings for NK cells, NK T cells were dramatically diminished in Vav-1(-/-) mice and splenocytes from Vav-1 mutant mice failed to produce IL-4 in response to in vivo CD3 stimulation. These data highlight the important role of Vav-1 in NK T cell development and NK cell function. PMID- 11500825 TI - A critical role for IL-12 in CCR5 induction on T cell receptor-triggered mouse CD4(+) and CD8(+) T cells. AB - Despite increasing evidence for the role of the chemokine system in leukocyte trafficking, the mechanism underlying the induction of chemokine receptors is poorly understood. Here, we investigated how CCR5, a chemokine receptor implicated in T cell migration to inflammatory sites, is induced in the T cell. CCR5 mRNA was hardly detected in resting T cells and marginally induced following T cell receptor (TCR) stimulation. However, TCR-triggered T cells expressed IL-12 receptor, and stimulation with recombinant IL-12 resulted in high levels of CCR5 expression on both CD4(+) and CD8(+) T cells. In contrast, IL-2 failed to up regulate CCR5 expression. The effect of IL-12 was selective to CCR5 because IL-12 did not up-regulate CXCR3 expression. Surface expression of CCR5 was shown by staining with anti-CCR5 monoclonal antibody. Stimulation of these CCR5-positive T cells with the relevant chemokine MIP-1 alpha elicited Ca(2+) influx, showing that IL-12-induced CCR5 is functional. These results indicate a critical role for IL-12 in the induction of CCR5 on TCR-triggered T cells. PMID- 11500826 TI - The Fas-FasL death receptor and PI3K pathways independently regulate monocyte homeostasis. AB - Peripheral blood-derived monocytes spontaneously undergo apoptosis mediated by Fas-Fas ligand (FasL) interactions. Activation of monocytes by LPS or TNF-alpha prevents spontaneous monocyte apoptosis through an unknown mechanism. Here, we demonstrate that LPS and TNF-alpha up-regulate Flip and suppress spontaneous Fas FasL mediated monocyte apoptosis and caspase 8 and 3 activation. Flip was responsible for this protection, since inhibition of Flip by antisense oligonucleotides in the presence of LPS or TNF-alpha restored monocyte sensitivity to spontaneous apoptosis. We also investigated whether the PI3K pathway contributes to the suppression of spontaneous monocyte apoptosis mediated by LPS and TNF-alpha. Monocytes treated with a reversible PI3K inhibitor (LY294002) displayed enhanced apoptosis, while LPS and TNF-alpha partially protected against apoptosis mediated by LY294002. However, direct suppression of Fas-FasL interactions by addition of neutralizing anti-FasL antibody did not further suppress LY294002-induced apoptosis in the presence of LPS or TNF-alpha. Collectively, these data demonstrate that LPS or TNF-alpha protect monocytes from death receptor-mediated apoptosis through the up-regulation of Flip, but not apoptosis initiated by inhibition of the PI3K pathway. PMID- 11500827 TI - Production of IL-4 and leukemia inhibitory factor by T cells of the cumulus oophorus: a favorable microenvironment for pre-implantation embryo development. AB - The nature and the functional activity of immunocytes present in the cumulus oophorus, a mass of cells surrounding the oocyte, were examined here for the first time. The cumuli oophorus were obtained from women who had taken part in an in vitro fertilization program and were suffering from blocked fallopian tubes. Both macrophages and CD4(+) T cells were detected in all cumuli. CD4(+) T cell clones, generated from T cells of these cumuli, showed higher potential to produce IL-4 and leukemia inhibitory factor (LIF) than CD4(+) T cell clones generated from peripheral blood or ovary specimens from the same women. More importantly, IL-4 and LIF, but not IFN-gamma mRNA was found to be constitutively expressed in vivo by cumulus oophorus cells. Progesterone is highly produced by the cumulus oophorus/oocyte complex. We recently showed that progesterone up regulates the production of LIF by T cells and that the progesterone-induced LIF production is mediated by IL-4. Progesterone produced by cumulus granulosa cells may favor IL-4 production by T cells, which in turn can produce LIF. As the treatment with LIF enhances the in vitro growth and development of mammalian embryos, our data suggest that T cells present in the cumulus oophorus produce cytokines that may provide a microenvironment suitable for pre-implantation development of the mammalian embryo. PMID- 11500828 TI - CD28 co-stimulates TCR/CD3-induced phosphoinositide turnover in human T lymphocytes. AB - Upon engagement of TCR with peptide-MHC complexes displayed on the surface of antigen-presenting cells, T lymphocytes undergo a sustained elevation of intracellular Ca(2+) concentration([Ca(2+)](i)), which is required for cytokine production. In the present work, we investigate how inositol lipid metabolism can be activated for a prolonged time to ensure a sustained link between receptor triggering and downstream signaling effectors. Four lines of evidence indicate that an extensive phosphoinositide turnover induced by TCR and CD28 engagement allows this task to be accomplished: (i) continuous phosphoinositide breakdown is required for a sustained [Ca(2+)](i )increase in antigen-stimulated T cells; (ii) TCR triggering results in a continuous release of inositol phosphates from the cell membrane paralleled by a massive and sustained phosphoinositide re-synthesis due to free inositol re-incorporation; (iii) TCR-induced phosphoinositide turnover is strongly increased by CD28 ligation; and (iv) CD28 engagement augments and sustains the TCR-induced [Ca(2+)](i )increase. Our results show that the T cell pool of phosphoinositides is continuously re-formed during T cell-APC cognate interaction, thereby explaining how sustained receptor triggering can transduce an equally sustained [Ca(2+)](i) increase. Importantly, our data identify a novel step in the signaling cascade where co-stimulation converges with TCR-generated signals to sustain and amplify the activation process. PMID- 11500829 TI - The role of MyD88 and TLR4 in the LPS-mimetic activity of Taxol. AB - Taxol can mimic bacterial lipopolysaccharide (LPS) by activating mouse macrophages in a cell cycle-independent, LPS antagonist-inhibitable manner. Macrophages from C3H/HeJ mice, which have a spontaneous mutation in Toll-like receptor 4 (TLR4), are hyporesponsive to both LPS and Taxol, suggesting that LPS and Taxol may share a signaling pathway involving TLR4. To determine whether TLR4 and its interacting adaptor molecule MyD88 are necessary for Taxol's LPS mimetic actions, we examined Taxol responses of primary macrophages from genetically defective mice lacking either TLR4 (C57BL/10ScNCr) or MyD88 (MyD88 knockout). When stimulated with Taxol, macrophages from wild-type mice responded robustly by secreting both TNF and NO, while macrophages from either TLR4-deficient C57BL/10ScNCr mice or MyD88 knockout mice produced only minimal amounts of TNF and NO. Taxol-induced NF-kappa B-driven luciferase activity was reduced after transfection of RAW 264.7 macrophages with a dominant negative version of mouse MyD88. Taxol-induced microtubule-associated protein kinase (MAPK) activation and NF-kappa B nuclear translocation were absent from TLR4-null macrophages, but were preserved in MyD88 knockout macrophages with a slight delay in kinetics. Neither Taxol-induced NF-kappa B activation, nor I kappa B degradation was affected by the presence of phosphatidylinositol 3-kinase inhibitors. These results suggest that Taxol and LPS not only share a TLR4/MyD88-dependent pathway in generating inflammatory mediators, but also share a TLR4-dependent/MyD88-independent pathway leading to activation of MAPK and NF-kappa B. PMID- 11500830 TI - Selective recruitment of Th2-type cells and evasion from a cytotoxic immune response mediated by viral macrophage inhibitory protein-II. AB - The viral CC chemokine macrophage inhibitory protein-II (vMIP-II) encoded by human herpes virus 8 (HHV-8) binds to multiple chemokine receptors, however, its ability to control the initial recruitment of specific leukocyte subtypes from the peripheral circulation has not been fully clarified. Here we show that vMIP II blocks the firm arrest and transmigration of monocytes or Th1-like T lymphocytes triggered by RANTES immobilized on activated human microvascular endothelium (HMVEC) under flow conditions. The internalization of the receptors CCR1 and CCR5 that mediate arrest and transmigration of these cells in response to RANTES was prevented by vMIP-II, supporting its role as an antagonist of CCR1 and CCR5. In contrast, vMIP-II triggered the firm arrest of eosinophils and Th2 like T cells by engaging CCR3, as confirmed by its down-regulation. Immunohistochemical analysis of HHV-8-associated Kaposi's sarcoma lesions marked by vMIP-II expression and mononuclear cell infiltration revealed a predominance of Th2-type CCR3(+) lymphocytes over Th1-type CXCR3(+)/CCR5(+) leukocytes, indicating that as a CCR3 agonist vMIP-II can drive a Th2-type immune response in vivo. Thus, our data provide evidence for a immunomodulatory role of vMIP-II in directing inflammatory cell recruitment away from a Th1-type towards a Th2-type response and thereby facilitating evasion from cytotoxic reactions. PMID- 11500831 TI - Impaired IgE response in SWAP-70-deficient mice. AB - Protein SWAP-70 was initially isolated from nuclei of activated B cells and was implicated in the immunoglobulin class switch process. After B cell activation the protein translocates from the cytoplasm to the nucleus, and may serve to signal nuclear processes. We have generated mice deficient in SWAP-70 and found three main differences when compared to wild-type mice: (i) their B lymphocytes are two- to threefold more sensitive to gamma-irradiation than B cells of wild type; (ii) SWAP-70-deficient mice developed autoantibodies at a much higher frequency; and (iii) the CD40 signaling pathway is compromised in the mutant mice. CD40-dependent switching to the IgE isotype is reduced five- to eightfold in vitro. In SWAP-70-deficient mice, IgE levels prior to immunization were six- to sevenfold lower than in wild-type mice, and after immunization three- to fourfold lower. CD40-induced proliferation was transiently increased in the mutant. LPS-induced switching to other isotypes, however, and LPS-induced proliferation were normal. We propose that SWAP-70 serves a specific role in the CD40 signaling pathway, in particular in the IgE response. PMID- 11500832 TI - Sampling of complementing self-antigen pools by thymic stromal cells maximizes the scope of central T cell tolerance. AB - Expression of peripheral antigens in the thymus has been implicated in T cell tolerance and autoimmunity, yet the identity of cells involved remains elusive. Here we show that antigen expression in a minor fraction of medullary thymic epithelial cells leads to deletion of specific CD4 T cells. Strikingly, this deletion is not dependent on cross-presentation by hemopoietic antigen-presenting cells, which have been ascribed a predominant role in negative selection. By contrast, when the same antigen enters the thymus via the blood stream, negative selection is strictly dependent on antigen presentation by hemopoietic cells. These findings imply that the (re)-presentation of "self" by thymic stromal cells is non-redundant, and that different thymic antigen-presenting cells instead cover complementing sets of self-antigens, thus maximizing the scope of central tolerance PMID- 11500834 TI - Laser capture microdissection-based expression profiling identifies PD1-ligand as a target of the nude locus gene product. AB - T cell development requires the interaction of developing thymocytes with thymic epithelial cells. Thymic epithelial cells acquire their unique phenotype under the control of the winged-helix transcription factor Whn, which is lacking in the nude mouse. Whn-dependent genes may therefore be important regulators of lympho epithelial interactions. To identify Whn target genes we isolated RNA populations of wild-type and nude thymic anlagen from embryonic day 12.5 embryos by laser capture microdissection and compared them by gene expression profiling on microarrays representing 22,000 transcripts. All cDNA with expression differences confirmed by quantitative RT-PCR using RNA from individual anlagen and by in situ hybridization were found to be present in wild-type and absent in nude samples. Three of eight confirmed transcripts were of hematopoietic origin; these transcripts emanate from hematopoietic precursors which have just entered the thymic anlage. Five transcripts were of epithelial origin; one of these corresponds to the recently identified PD-1 ligand (PD-L1), the receptor of which is known to modulate positive selection and to play a role in the control of autoimmunity, and the remaining transcripts code for novel genes. The presented results support our prediction that this systematic approach by gene expression profiling yields regulators of thymopoiesis. PMID- 11500833 TI - Weak TCR stimulation induces a calcium signal that triggers IL-4 synthesis, stronger TCR stimulation induces MAP kinases that control IFN-gamma production. AB - Th1 and Th2 cells produce different cytokines and have distinct functions. Th1/Th2 cell differentiation is influenced, among other factors, by the nature of TCR-MHC interactions. However, how the TCR transduces a signal resulting in IFN gamma or IL-4 production is a matter of debate. For example, some authors reported a loss of calcium signaling pathway in Th2 cells. We used a T cell hybridoma producing IL-4 upon weak TCR stimulation and both IL-4 and IFN-gamma for strong TCR engagement as a model to study how TCR signaling pathways are differentially activated in both conditions of stimulation and how this influences the production of cytokines. We show that: (1) the calcium response is identical following weak and strong TCR stimulation; (2) mitogen-activated protein kinase(MAPK) activation is a gradual phenomenon depending upon the strength of TCR activation; (3) a calcium response, even weak, triggers IL-4 expression; (4) IFN-gamma synthesis requires not only a calcium response but also MAPK activation. The MAPK pathway is dispensable for IL-4 production, although it amplifies IL-4 synthesis upon strong TCR stimulation; (5) TCR-induced IL-4 production also depends on calcium signaling in Th2 cells, while IFN-gamma synthesis is dependent, in addition, on MAPK activation in Th1 cells. PMID- 11500835 TI - The membrane-bound but not the soluble form of human Fas ligand is responsible for its inflammatory activity. AB - The ectopic expression of Fas ligand (FasL/CD95L) in tissues or tumors induces neutrophil infiltration and the destruction of the tissues or the rejection of tumors. It has been suggested that the infiltrated neutrophils are responsible for the latter phenomena. FasL is synthesized as a type II transmembrane protein, and soluble FasL is produced by a proteolytic mechanism from the membrane-bound form. We previously demonstrated that uncleavable membrane-bound FasL of mice induces IL-1 beta release from inflammatory cells, and suggested that the IL-1 beta enhances neutrophil infiltration. However, recent papers reported that human soluble FasL is directly chemoattractive to neutrophils in vitro and proposed that the soluble form of FasL is responsible for its inflammatory activity. Therefore, in this report, we investigated which form is responsible for the inflammatory activities of human FasL. We produced tumor cell lines expressing one or both forms of human FasL. Cells expressing both forms or only the membrane bound form of FasL induced neutrophil infiltration when transplanted into the peritoneal cavity of syngeneic mice, while cells expressing only the soluble form did not. Purified soluble FasL failed to induce neutrophil infiltration in vivo. IL-1 beta release from inflammatory peritoneal exudate and acceleration of tumor rejection were also mediated by membrane-bound but not soluble FasL. These results indicate that the membrane-bound form of FasL is primarily responsible for its inflammatory activity. PMID- 11500836 TI - CD4(+)CD8(dim) T lymphocytes exhibit enhanced cytokine expression, proliferation and cytotoxic activity in response to HCMV and HIV-1 antigens. AB - CD4(+)CD8(dim) T cells represent a minor subset of the total CD3(+) T cell population in peripheral blood. Although transient and persistent expansions of these cells have been reported in both healthy and diseased individuals, the functional properties of the CD4(+)CD8(dim) population are largely unknown. In this study, we examined antigen-specific cytokine and proliferative responses of the CD4(+)CD8(dim) subset. In whole blood cultures stimulated with the viral antigens HCMV and HIV-1, a significant fraction of the CD4(+)CD8(dim) subset exhibited cytokine expression and proliferation in response to antigen activation. Typically, the CD4(+)CD8(dim) population contained two- to eightfold higher frequencies of antigen-specific cytokine producing cells than the CD4(+)CD8(-) population. Phenotypic analysis of the cytokine expressing CD4(+)CD8(dim) population indicated that these cells are memory T cells, with a high frequency of this population expressing the cytotoxic markers CD56 and perforin. Furthermore, the CD4(+)CD8(dim) cytokine responses to CMV were shown to be MHC class II dependent. Significantly, purified CD4(+)CD8(dim) T cells were found to possess higher CMV-specific cytotoxic activity than purified CD4(+)CD8( ) T cells in a standard (51)Cr-release CTL assay. Thus, CD4(+)CD8(dim) T cells appear to be MHC class II dependent, are capable of cytolytic effector activity, and are highly enriched within the CD4(+) cell populations specific for HCMV and HIV-1. PMID- 11500837 TI - Cross reactivity of three T cell attracting murine chemokines stimulating the CXC chemokine receptor CXCR3 and their induction in cultured cells and during allograft rejection. AB - Recent work identified the murine gene homologous to the human T cell attracting chemokine CXC receptor ligand 11 (CXCL11, also termed I-TAC, SCYB11, ss-R1, H174, IP-9). Here, the biological activity and expression patterns of murine CXCL11 relative to CXCL9 (MIG) and CXCL10 (IP-10/crg-2), the other two CXCR3 ligands, were assessed. Calcium mobilization and chemotaxis experiments demonstrated that murine CXCL11 stimulated murine CXCR3 at much lower doses than murine CXCL9 or murine CXCL10. Murine CXCL11 also evoked calcium mobilization in CHO cells transfected with human CXCR3 and was chemotactic for CXCR3-expressing human T lymphocytes as well as for 300--19 pre-B cells transfected with human or murine CXCR3. Moreover, murine CXCL11 blocked the chemotactic effect of human CXCL11 on human CXCR3 transfectants. Depending on cell type (macrophage-like cells RAW264.7, J774A.1, fetal F20 and adult dermal fibroblasts, immature and mature bone marrow-derived dendritic cells) and stimulus (interferons, LPS, IL-1 beta and TNF-alpha), an up to 10,000-fold increase of CXCL9, CXCL10 and CXCL11 mRNA levels, quantified by real-time PCR, was observed. In vivo, the three chemokines are constitutively expressed in various tissues from healthy BALB/c mice and were strongly up-regulated during rejection of allogeneic heart transplants. Chemokine mRNA levels exceeded those of CXCR3 and IFN-gamma which were induced with similar kinetics by several orders of magnitude. PMID- 11500838 TI - Pathfinding by sensory axons in Drosophila: substrates and choice points in early lch5 axon outgrowth. AB - We have examined the pattern of axon growth from the lateral chordotonal (lch5) neurons in the body wall of the Drosophila embryo and identified cellular substrates and choice points involved in early axon pathfinding by these sensory neurons. At the first choice point (TP1), the lch5 growth cones contact the most distal cells of the spiracular branch (SB) of the trachea. The SB provides a substrate along which the axons extend internally to the level of the intersegmental nerve (ISN). In the absence of the SB, the lch5 axons often stall near TP1 or follow aberrant routes towards the CNS. At the second choice point (TP2), the lch5 growth cones make their first contact with other axons and turn ventrally toward the CNS, fasciculating specifically with the motor axons of the ISN. PMID- 11500839 TI - Testosterone treatment increases the metabolic capacity of adult avian song control nuclei. AB - In songbirds, the size of brain nuclei that control song learning and production change seasonally. These changes are mainly controlled by seasonal changes in plasma testosterone (T) concentration. One hypothesis to explain why it may be adaptive for these areas to regress in the fall is that this would decrease the metabolic demand of maintaining a large song system when singing is reduced or absent. We used a marker for cellular metabolism to examine birds with regressed song nuclei and compared them to birds whose song nuclei were induced to grow by administration of exogenous T. Photorefractory male Gambel's white-crowned sparrows were captured during their autumnal migration and kept in outdoor aviaries on a natural photoperiod. We implanted birds with Silastic capsules containing T or with empty implants. Three weeks later the birds were sacrificed. We assayed the brains for cytochrome oxidase (CO) activity and measured the volume of four song nuclei: HVc, RA, 1MAN, and area X. All four nuclei increased in volume in response to T treatment. T treatment increased the metabolic capacity of area X, HVc, and RA relative to surrounding tissue but had no effect on the metabolic capacity of 1MAN. These results support the hypothesis that song nuclei are more metabolically active under the influence of T than they are when plasma T levels are low. PMID- 11500840 TI - The fates of cells generated at the end of neurogenesis in developing mouse cortex. AB - Most cerebral cortical neurons are generated between embryonic days 11 and 17 (E11-17) in the mouse. Radial glial cells also proliferate during this time; they can give rise to neurons and many later transform into astrocytes. It is thought that most glial cells comprising the mature cortex, including additional astrocytes, are generated after neurogenesis is complete. Little is known about the cellular events that occur during the transition from the phase dominated by neurogenesis to that of gliogenesis. We labeled cells generated on E18 and E19 and the day of birth (P0) with bromodeoxyuridine and followed their fates over the following 20 days. Our results showed that, on E18-P0, cells divide throughout the ventricular zone, subventricular zone, intermediate zone, and to a lesser extent, the developing cortical plate, whereas neuronal precursors generated prior to E18 divide in the ventricular zone. Our results indicated that 30-40% of cells dividing on E18 give rise to neurons that migrate to the most superficial part of the cortex. The rest of the cells dividing on E18 and 76-94% of cells generated on E19 and P0 express the QKI RNA-binding protein, indicating that they either remain as multipotential progenitors or develop into glial cells. Nine to fifteen percent of cells generated on E18-P0 become glial fibrillary acidic protein-positive astrocytes. Many E19 and P0 labeled cells disappear between 2 and 20 days postlabeling, probably because they continue to divide. We conclude that the population of cells produced at the end of cortical neurogenesis is heterogeneous and comprises postmitotic neurons, glia (including astrocytes), and possibly multipotential progenitors. PMID- 11500841 TI - Transneuronal tracing from sympathectomized lumbar epaxial muscle in female rats. AB - Pseudorabies virus (PRV) has been used as a transneuronal tracer to study central neural networks, including the central control of the lordosis-producing, lumbar epaxial muscles. Within muscles, however, the sympathetic innervation of blood vessels poses a confounding source of tracer labeling in the CNS. The present study destroyed sympathetic nerves before injection of PRV, thereby allowing for a more selective uptake by somatic motoneurons. Specifically, a focal sympathectomy was created by the injection of dopamine-beta-hydroxylase immunotoxin (DHIT). When PRV was injected into control rats, both somatic motoneurons within the ventral horn of the spinal cord and sympathetic preganglionic neurons within the intermediolateral column (IML) of the spinal cord became labeled. Additionally, labeled neurons were observed in many brain regions, including those previously implicated in the control of the lordosis reflex (e.g., the medullary reticular formation; MRF) and those previously implicated in the control of vasomotor tone (e.g., the rostral ventrolateral medulla; RVLM). When injected into DHIT-pretreated animals, PRV labeling in ventral horn neurons persisted in many animals; however, labeling in IML was eliminated in almost every case. In these animals, PRV labeling was absent in brain areas traditionally associated with vasomotor tone, such as RVLM, whereas labeling persisted in brain areas previously implicated in the control of the lordosis response, such as MRF. The results support the connectivity of spinal and medullary structures with the somatic control of the lordosis-producing muscles and provide a more detailed description of these portions of the putative lordosis-relevant neurocircuitry. PMID- 11500842 TI - GABA exerts opposite influence on warm and cold sensitive neurons in medial preoptic area in rats. AB - The preoptic area regulates body temperature. GABA-ergic terminals and receptors are present in this area. Local microinjection studies have shown that GABA, its agonist, and its antagonist in this area may modulate body temperature. However, there are warm and cold sensitive neurons, and they are known to be affected by local and peripheral temperatures. In order to understand the mechanism of action of GABA in temperature regulation at the cellular level it was necessary to study the effect of GABA on individual thermosensitive neurons in in vivo preparations. Hence, in this study the responses of preoptic area thermosensitive and insensitive neurons to microiontophoretic application of picrotoxin, a GABA-A antagonist, were studied in anaesthetized rats. It was observed that a majority of both the thermosensitive and insensitive neurons were affected by microiontophoretic application of picrotoxin. Although almost an equal number of cold and warm sensitive neurons were affected, a majority of the cold sensitive neurons were excited, while a majority of the warm sensitive neurons were inhibited by picrotoxin. The results suggested that in normal conditions GABA acts through GABA-A receptor in modulating the spontaneous activity of thermosensitive neurons in the preoptic area. Furthermore, the results of the present study taken together with other reports suggest that normally GABA exerts a direct inhibitory action on the cold sensitive neurons, while it acts on presynaptic heteroreceptors, possibly on norepinephrinergic afferent input terminals on the warm sensitive neurons, for mediating its action. PMID- 11500843 TI - Ovariectomy attenuates dendritic growth in hormone-sensitive spinal motoneurons. AB - The lumbar spinal cord of rats contains the sexually dimorphic, steroid-sensitive spinal nucleus of the bulbocavernosus (SNB). Dendritic development of SNB motoneurons in male rats is biphasic, initially showing exuberant growth through 4 weeks of age followed by a retraction to mature lengths by 7 weeks of age. The initial growth is steroid dependent, attenuated by castration or aromatase inhibition, and supported by hormone replacement. Dendritic retraction is also steroid sensitive and can be prevented by testosterone treatment, but is unaffected by aromatase inhibition. Together, these results suggest a role for estrogens during the initial growth phase of SNB development. In this study, we tested whether ovarian hormones could support SNB somal and dendritic development. Motoneuron morphology was assessed in normal males and in females perinatally masculinized with dihydrotestosterone and then either ovariectomized or left intact. SNB motoneurons were retrogradely labeled with cholera toxin-HRP at 4 or 7 weeks of age and reconstructed in three dimensions. Initial growth of SNB dendrites was reduced after ovariectomy in masculinized females. However, no differences in dendritic length were seen at 7 weeks of age between intact and ovariectomized masculinized females, and lengths in both groups were significantly lower than those of normal males. Together with previous findings, these results suggest that estrogens are involved in the early growth of SNB dendrites, but not in their subsequent retraction. PMID- 11500844 TI - Short- and long-chain natural polyamines play specific roles in adult cricket neuroblast proliferation and neuron differentiation in vitro. AB - In the house cricket (Acheta domesticus) mushroom bodies, neurogenesis still occurs during adulthood. Using in vitro approaches, the respective roles of natural polyamines in neurogenesis were examined. Mushroom body neuroblast proliferation was assayed in organotypic culture using 5-bromo, 2'-deoxyuridine labeling. The number of labeled cells was significantly increased when putrescine was added to culture medium, whereas spermidine and spermine supplementation did not alter cell proliferation. Conversely, in vitro morphometric studies on mushroom body neurons cultured in a defined medium showed that putrescine addition failed to alter any morphological character of these interneurons, whereas addition of the long-chain polyamines, spermidine and spermine, stimulated neuron differentiation. These two polyamines significantly increased total neurite length; moreover, spermidine-treated cells exhibited more branches than the controls. The present data demonstrate that putrescine has a mitogenic effect on mushroom body neuronal precursors, and that spermidine and spermine, which failed to induce neuroblast proliferation, act on neuronal differentiation, inducing neurite outgrowth. Our results indicate that short- and long-chain polyamines play specific roles during neurogenesis, and provide a basis for further studies on neuronal precursor proliferation and differentiation. PMID- 11500845 TI - Evaluation of confocal microscopy system performance. AB - BACKGROUND: The confocal laser scanning microscope (CLSM) has been used by scientists to visualize three-dimensional (3D) biological samples. Although this system involves lasers, electronics, optics, and microscopes, there are few published tests that can be used to assess the performance of this equipment. Usually the CLSM is assessed by subjectively evaluating a biological/histological test slide for image quality. Although there is a use for the test slide, there are many other components in the CLSM that need to be assessed. It would be useful if tests existed that produced reference values for machine performance. The aim of this research was to develop quality assurance tests to ensure that the CLSM was stable while delivering reproducible intensity measurements with excellent image quality. METHODS: Our ultimate research objective was to quantify fluorescence using a CLSM. To achieve this goal, it is essential that the CLSM be stable while delivering known parameters of performance. Using Leica TCS-SP1 and TCS-4D systems, a number of tests have been devised to evaluate equipment performance. Tests measuring dichroic reflectivity, field illumination, lens performance, laser power output, spectral registration, axial resolution, laser stability, photomultiplier tube (PMT) reliability, and system noise were either incorporated from the literature or derived in our laboratory to measure performance. These tests are also applicable to other manufacturer's systems with minor modifications. RESULTS: A preliminary report from our laboratory has addressed a number of the QA issues necessary to achieve CLSM performance. This report extends our initial work on the evaluation of CLSM system performance. Tests that were described previously have been modified and new tests involved in laser stability and sensitivity are described. The QA tests on the CLSM measured laser power, PMT function, dichroic reflection, spectral registration, axial registration, system noise and sensitivity, lens performance, and laser stability. Laser power stability varied between 3% and 30% due to various factors, which may include incompatibility of the fiber-optic polarization with laser polarization, thermal instability of the acoustical optical transmission filter (AOTF), and laser noise. The sensitivity of the system was measured using a 10-microm Spherotech bead and the PMTs were assessed with the CV concept (image noise). The maximum sensitivity obtainable on our TCS-SP1 system measured on the 10-microm Spherotech beads was approximately 4% for 488 nm, 2.5% for 568 nm, 20% for 647 nm, and 19% for 365 nm laser light. The values serve as a comparison to test machine sensitivity from the same or different manufacturers. CONCLUSIONS: QA tests are described on the CLSM to assess performance and ensure that reproducing data are obtained. It is suggested strongly that these tests be used in place of a biological/histological sample to evaluate system performance. The tests are more specific and can recognize instrument functionality and problems better than a biological/histological sample. Utilization of this testing approach will eliminate the subjective assessment of the CLSM and may allow the data from different machines to be compared. These tests are essential if one is interested in making intensity measurements on experimental samples as well as obtaining the best signal detection and image resolution from a CLSM. Published 2001 Wiley-Liss, Inc. PMID- 11500846 TI - Statistical evaluation of confocal microscopy images. AB - BACKGROUND: The coefficient of variation (CV) is defined as the standard deviation (sigma) of the fluorescent intensity of a population of beads or pixels expressed as a proportion or percentage of the mean (mu) intensity (CV = sigma/mu). The field of flow cytometry has used the CV of a population of bead intensities to determine if the flow cytometer is aligned correctly and performing properly. In a similar manner, the analysis of CV has been applied to the confocal laser scanning microscope (CLSM) to determine machine performance and sensitivity. METHODS: Instead of measuring 10,000 beads using a flow cytometer and determining the CV of this distribution of intensities, thousands of pixels are measured from within one homogeneous Spherotech 10-microm bead. Similar to a typical flow cytometry population that consists of 10,000 beads, a CLSM scanned image consists of a distribution of pixel intensities representing a population of approximately 100,000 pixels. In order to perform this test properly, it is important to have a population of homogeneous particles. A biological particle usually has heterogeneous pixel intensities that correspond to the details in the biological image and thus shows more variability as a test particle. RESULTS: The bead CV consisting of a population of pixel intensities is dependent on a number of machine variables that include frame averaging, photomultiplier tube (PMT) voltage, PMT noise, and laser power. The relationship among these variables suggests that the machine should be operated with lower PMT values in order to generate superior image quality. If this cannot be achieved, frame averaging will be necessary to reduce the CV and improve image quality. There is more image noise at higher PMT settings, making it is necessary to average more frames to reduce the CV values and improve image quality. The sensitivity of a system is related to system noise, laser light efficiency, and proper system alignment. It is possible to compare different systems for system performance and sensitivity if the laser power is maintained at a constant value. Using this bead CV test, 1 mW of 488 nm laser light measured on the scan head yielded a CV value of 4% with a Leica TCS-SP1 (75-mW argon-krypton laser) and a CV value of 1.3% with a Zeiss 510 (25-mW argon laser). A biological particle shows the same relationship between laser power, averaging, PMT voltage, and CV as do the beads. However, because the biological particle has heterogeneous pixel intensities, there is more particle variability, which does not make as useful as a test particle. CONCLUSIONS: This CV analysis of a 10-microm Spherotech fluorescent bead can help determine the sensitivity in a confocal microscope and the system performance. The relationship among the factors that influence image quality is explained from a statistical endpoint. The data obtained from this test provides a systematic method of reducing noise and increasing image clarity. Many components of a CLSM, including laser power, laser stability, PMT functionality, and alignment, influence the CV and determine if the equipment is performing properly. Preliminary results have shown that the bead CV can be used to compare different confocal microscopy systems with regard to performance and sensitivity. The test appears to be analogous to CV tests made on the flow cytometer to assess instrument performance and sensitivity. Published 2001 Wiley Liss, Inc. PMID- 11500847 TI - A standard for calibration and shading correction of a fluorescence microscope. AB - BACKGROUND: Numerous applications of fluorescence microscopy require quantitation of signal intensity in reproducible units. Two problems must be overcome to achieve this goal. First, due to various instrumental factors, the same sample imaged on two microscopes or even on the same microscope at different times may produce highly divergent readings. Second, because of shading, some areas within the same field may appear brighter than others despite the same amount of fluorophore. The first type of variability requires calibration using a sample of reproducible fluorescence yield; to correct for shading, a uniform fluorescent field is needed. METHODS: Standard slides were prepared by placing several microliters of 10%-50% w/v fluorescein or rhodamine between a coverglass and a slide. They were used to perform shading correction and normalization under a variety of imaging conditions. RESULTS: Concentrated fluorophores produced a uniform fluorescent field of moderate and reproducible brightness. By expressing the staining of a biological object in the units of standard slides, identical results were obtained irrespective of the imaging conditions or the microscope used. We compared shading correction based on concentrated fluorescein with two other standards. Concentrated fluorescein resulted in the best equalization of the field. CONCLUSIONS: Standardization of fluorescent images can be achieved by normalizing them to the image of a concentrated solution of a fluorophore. Due to its simplicity and efficiency, this method can be used in clinical analysis as well as in routine laboratory practice. PMID- 11500848 TI - Optimization of DOP-PCR amplification of DNA for high-resolution comparative genomic hybridization analysis. AB - BACKGROUND: Whole-genome amplification of minute samples of DNA for the use in comparative genomic hybridization (CGH) analysis has found widespread use, but the method has not been well validated. METHODS: Four protocols for degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) and fluorescence labeling were applied to test DNA from normal and K-562 cells. The DNA products were used for CGH analysis. RESULTS: The DOP-PCR-amplified DNA from each protocol produced hybridizations with different qualities. These could be seen primarily as differences in background staining and signal-to-noise ratios, but also as characteristic deviations of normal/normal hybridizations. One DOP-PCR-protocol was further investigated. We observed concordance between CGH results using unamplified and DOP-PCR-amplified DNA. An example of an analysis of an invasive carcinoma of the breast supports the practical value of this approach. CONCLUSIONS: DOP-PCR-amplified DNA is applicable for high- resolution CGH, the results being similar to those of CGH using unamplified DNA. PMID- 11500849 TI - Multiplexed molecular interactions of nuclear receptors using fluorescent microspheres. AB - BACKGROUND: We describe a novel microsphere-based system to identify and characterize multiplexed interactions of nuclear receptors with peptides that represent the LXXLL binding region of coactivator proteins. METHODS: In this system, individual microsphere populations with unique red and orange fluorescent profiles are coupled to specific coactivator peptides. The coactivator peptide coupled microsphere populations are combined and incubated with a nuclear receptor that has been coupled to a green fluorochrome. Flow cytometric analysis of the microspheres simultaneously decodes each population and detects the binding of receptor to respective coactivator peptides by the acquisition of green fluorescence. RESULTS: We have used this system to determine the binding affinities of human estrogen receptor beta ligand binding domain (ERbeta LBD) and human peroxisome proliferator activated receptor gamma ligand binding domain (PPARgamma LBD) to a set of 34 coactivator peptides. Binding of ERbeta LBD to a coactivator peptide sequence containing the second LXXLL motif of steroid receptor coactivator-1 (SRC-1(2) (676-700) is shown to be specific and saturable. Analysis of receptor binding to a multiplexed set of coactivator peptides shows PPARgamma LBD binds with high affinity to cAMP response element binding protein (CBP) peptides and to the related P300 peptide while ERbeta LBD exibits little binding to these peptides. Using the microsphere-based assay we demonstrate that ERbeta LBD and PPARgamma LBD binding affinities for the coactivator peptides are increased in the presence of agonist (estradiol or GW1929, respectively) and that ERbeta LBD binding is decreased in the presence of antagonist (raloxifene or tamoxifen). CONCLUSIONS: This unique microsphere-based system is a sensitive and efficient method to simultaneously evaluate many receptor-coactivator interactions in a single assay volume. In addition, the system offers a powerful approach to study small molecule modulation of nuclear receptor binding. PMID- 11500850 TI - Epidermal growth factor receptor, c-erbB2 and c-erbB3 receptor interaction, and related cell cycle kinetics of SK-BR-3 and BT474 breast carcinoma cells. AB - BACKGROUND: Receptors belonging to the epidermal growth factor receptor (EGFR) family transfer extracellular signals by homotypic and heterotypic receptor interaction and cross-activation. Cell differentiation, death, and proliferation are regulated via these receptor-tyrosine-kinases. However, the initial mechanisms that lead to signal specificity and diversity, which cause a defined cellular response, are incompletely understood. We investigated the recruitment of receptor complexes in two c-erbB2-overexpressing breast carcinoma cell lines, SK-BR-3 and BT474, after ligand binding and its effects on intracellular signal transduction and cell cycle regulation. METHODS: In order to analyze the coaggregation of receptors on the cell surface induced by specific growth factor treatment, we used the flow cytometric Foerster-type fluorescence resonance energy transfer (FRET) technique. Cell cycle kinetics were monitored flow cytometrically via the anti-BrdU technique and acitivation of intracellular signal cascades was analyzed by Western blotting. RESULTS: After stimulation with EGF BT474, but not SK-BR-3, cells formed EGFR/c-erbB2 receptor complexes. Neither EGF nor heregulin (HRG) induced c-erbB2/c-erbB3 receptor complexes in BT474. However, SK-BR-3 cells exhibited a high amount of c-erbB2/c-erbB3 heterodimers even without growth factor stimulation which could be elevated after prolonged EGF and HRG treatment. In both cell lines, mitogen-activated protein kinase (MAPK) phosphorylation was detectable after short-term and prolonged EGF and HRG treatment. However, only SK-BR-3 cells showed a constitutive activation of both protein kinase B (PKB)/Akt and MAPK signaling pathways. Growth factor treatment caused an amplified PKB/Akt activation in this cell line. The induction of EGFR/c erbB2 complexes in BT474 was associated with shortening of the G1-phase of the cell cycle. In contrast, the concurrent activation of MAPK and PKB/Akt by EGF treatment led to an inhibition of proliferation in SK-BR-3 and can be attributed to missing EGFR/c-erbB2 heterodimers. HRG was a strong stimulator of proliferation in both cell lines. CONCLUSIONS: We show that in the presence of identical amounts of c-erbB2 receptors, the ligand-induced cellular response differs significantly. These differences were mediated by variances in signal transduction, most likely due to different recruitment of heterotypic receptor complexes. Overall, there is strong evidence that c-erbB2 receptor overexpression in breast cancer cells is an insufficient marker to determine cellular response in terms of cell proliferation. 2001. PMID- 11500851 TI - Effects of paclitaxel on cultured synovial cells from patients with rheumatoid arthritis. AB - BACKGROUND: Proliferation of synovial cells is considered to play a key role in rheumatoid arthritis (RA). Using paclitaxel, a unique antineoplastic agent known to suppress collagen-induced arthritis, we conducted an in vitro study of cell kinetics on cultured synovial cells from patients with RA. METHODS: Alterations of the cell cycle of cultured fibroblast-like synovial cells (FLSs) from patients with RA were studied using flow cytometry and laser scanning cytometry. Apoptosis and accumulation of cyclin concerning effects of paclitaxel were detected. RESULTS: Paclitaxel induced arrest of the cell cycle at G2/M phase and apoptosis in FLSs. The late stage of apoptosis was determined by the positivity of terminal deoxynucleotidyl transferase assay. Morphological observation by combined usage of both annexin V and propidium iodide on FLSs on a slide glass showed early apoptotic changes in detail. FLSs arrested at G2/M phase showed marked accumulation of cyclin B1. The effects of paclitaxel decreased on FLSs, which diminished proliferative activity. CONCLUSIONS: These data indicate that paclitaxel induces cell arrest at G2/M phase followed by apoptosis in human FLSs, which have high proliferative activity, and possible therapeutic effects of paclitaxel on RA. PMID- 11500852 TI - "Liquidless" cell staining by dye diffusion from gels and analysis by laser scanning cytometry: potential application at microgravity conditions in space. AB - BACKGROUND: Conventional staining of cells or tissue sections on microscope slides involves immersing the slides into solutions of dyes then rinsing to remove the unbound dye. There are instances, however, when use of stain solutions is undesirable-e.g., at microgravity conditions in space, where the possibility of accidental spill (many dyes are known carcinogens) introduces health hazard. Likewise, transporting bulk of liquid stains and rinses may be burdensome in certain situations such as field expeditions or combat. METHODS: The "liquidless" staining procedure is proposed in which the dyes are contained in thin strips of hydrated polyacrylamide or gelatin gels that have been presoaked in the stain solutions. Fluorochromes that have affinity to DNA (propidium iodide, PI; 4,6 diamidino-2-phenylindole, DAPI, Hoechst 33342) or to protein (sulforhodamine 101) were used to saturate the gels. The gel strips were placed over the prefixed cells or tissue sections deposited on microscope slides and relatively low (20 g/cm2) pressure was applied to ensure the contact. The cells were also stained by using commercially available mounting media into which DAPI or PI were admixed. Intensity of fluorescence of the PI stained cells was measured by laser scanning cytometry (LSC). RESULTS: Satisfactory cell and tissue staining, with minimal background, was achieved after 10-20 min contact between the cells and gels. Optimal concentrations of the dyes in the solutions used to presoak the gels was found to be 2-4-fold higher than the concentrations used routinely in cytometry. The measurements of intensity of cellular fluorescence by LSC revealed that the staining of DNA was stoichiometric as reflected by the characteristic cellular DNA content frequency histograms with distinct G1, S, and G2/M cell populations and 2:1 ratio of G2/M to G1 peak fluorescence. Individual gels can be saturated with more than a single dye-e.g., to obtain differential DNA and protein staining. Cell staining with DAPI or PI in the gelatin-based mounting media led to high fluorescence background while staining with DAPI in "aqueous" medium was satisfactory. CONCLUSIONS: Relatively fast staining of cells or tissue sections on microscope slides can be achieved by nonconvective dye diffusion using hydrated gels permeated with the dyes, applied to cells at low pressure. The quality of the staining provided by this methodology is comparable to conventional cell staining in dye solutions. PMID- 11500853 TI - Fluorescence resonance energy transfer analysis of cell surface receptor interactions and signaling using spectral variants of the green fluorescent protein. AB - BACKGROUND: Fluorescence resonance energy transfer (FRET) is a powerful technique for measuring molecular interactions at Angstrom distances. We present a new method for FRET that utilizes the unique spectral properties of variants of the green fluorescent protein (GFP) for large-scale analysis by flow cytometry. METHODS: The proteins of interest are fused in frame separately to the cyan fluorescent protein (CFP) or the yellow fluorescent protein (YFP). FRET between these differentially tagged fusion proteins is analyzed using a dual-laser FACSVantage cytometer. RESULTS: We show that homotypic interactions between individual receptor chains of tumor necrosis factor receptor (TNFR) family members can be detected as FRET from CFP-tagged receptor chains to YFP-tagged receptor chains. Noncovalent molecular complexation can be detected as FRET between fusions of CFP and YFP to either the intracellular or extracellular regions of the receptor chains. The specificity of the assay is demonstrated by the absence of FRET between heterologous receptor pairs that do not biochemically associate with each other. Interaction between a TNFR-like receptor (Fas/CD95/Apo 1) and a downstream cytoplasmic signaling component (FADD) can also be demonstrated by flow cytometric FRET analysis. CONCLUSIONS: The utility of spectral variants of GFP in flow cytometric FRET analysis of membrane receptors is demonstrated. This method of analyzing FRET allows probing of noncovalent molecular interactions that involve both the intracellular and extracellular regions of membrane proteins as well as proteins within the cells. Unlike biochemical methods, FRET allows the quantitative determination of noncovalent molecular associations at Angstrom level in living cells. Moreover, flow cytometry allows quantitative analyses to be carried out on a cell-by-cell basis on large number of cells. Published 2001 Wiley-Liss, Inc. PMID- 11500854 TI - T-cell receptor analysis by flow cytometry. PMID- 11500855 TI - Fluorescence spectroscopy of epithelial tissue throughout the dysplasia-carcinoma sequence in an animal model: spectroscopic changes precede morphologic changes. AB - BACKGROUND AND OBJECTIVE: The hamster cheek pouch carcinogenesis model, using chronic treatments of dimethylbenz[alpha]anthracene (DMBA) was used as a model system to investigate changes in epithelial tissue autofluorescence throughout the dysplasia-carcinoma sequence. STUDY DESIGN/MATERIALS AND METHODS: Fluorescence emission spectra were measured weekly from 42 DMBA-treated animals and 20 control animals at 337, 380, and 460 nm excitation. A subset of data in which histopathology was available was used to develop diagnostic algorithms to separate neoplastic and non-neoplastic tissue. The change in fluorescence intensity over time was examined in all samples at excitation-emission wavelength pairs identified as diagnostically useful. RESULTS: Algorithms based on autofluorescence can separate neoplastic and non-neoplastic tissue with 95% sensitivity and 93% specificity. Greatest contributions to diagnostic algorithms are obtained at 380 nm excitation, and 430, 470, and 600 nm emission. Changes in fluorescence intensity are apparent as early as 3 weeks after initial treatment with DMBA, whereas morphologic changes associated with dysplasia occur on average at 7.5-12.5 weeks after initial treatment. CONCLUSIONS: Fluorescence spectroscopy provides a potential tool to identify biochemical changes associated with dysplasia and hyperplasia, which precede morphologic changes observed in histologically stained sections. PMID- 11500856 TI - Fluorescent imaging in a glioma model in vivo. AB - BACKGROUND AND OBJECTIVE: Nile blue dyes have been shown to have affinity for tumor tissue as compared to surrounding normal tissue and to be relatively non toxic. We have employed EtNBA, a lipophilic, fluorescent benzophenoxazine dye, in a murine model to image subcutaneous and intracranial U-87 glioma implants. STUDY DESIGN/MATERIALS AND METHODS: The imaging system used to detect fluorescence consists of a SIT video camera fitted with a zoom microscope-magnifying lens. The tumor was illuminated with a 632.8-nm diffuse beam from a helium-neon laser. The video image was processed using a Sony image processor to give real-time pseudocolor and enhanced black and white images. RESULTS: Following subcutaneous injection of the dye at doses of 2.5-5.0 mg/kg bw, we observed a gradual increase of the fluorescent signal from the tumor which peaked 1-3 hours post-injection with variable selectivity (typically 4:1) for tumor to normal surrounding tissues permitting the clear demarcation of the tumor. CONCLUSIONS: The present in vivo study demonstrates that EtNBA is a safe and effective photodiagnostic agent, able to demarcate U87-MG solid tumors in mice on a real-time basis at a concentration of 2.5-5.0 mg/kg 1-3 hours after administration. PMID- 11500857 TI - Near-infrared photo-excited emission from tissues treated at different temperature levels. AB - BACKGROUND AND OBJECTIVE: There is a lack of methods to evaluate the extent of thermal treatment of biological tissue. The intensity of the near-infrared (NIR) emission photo-excitation was investigated from tissue undergoing different levels of heat treatment. STUDY DESIGN/MATERIALS AND METHODS: Chicken muscle was heated in an oven at different temperature levels ranging from 40 degrees C until burn-off. The spectral emission intensity from these heat-treated tissues was measured with a CCD camera and the intensity was calculated. RESULTS: The emission intensity increased proportionally with respect to the extent of treatment temperature until burning. Linear relationships between treatment temperature and the emission intensity from tissue samples were found in three temperature ranges: from 40 to 160 degrees C, from 165 to 220 degrees C, and from 225 to 250 degrees C. CONCLUSIONS: The change in tissue damage after heat treatment could be detected by measuring the NIR emission intensity from the thermally damaged tissues. PMID- 11500858 TI - Treatment of canine hemangiopericytomas with photodynamic therapy. AB - BACKGROUND AND OBJECTIVES: Canine hemangiopericytomas are a commonly occurring neoplasm with a clinical course of recurrence after surgical removal. This study sought to evaluate Photochlor (HPPH) photodynamic therapy (HPPH-PDT) as an adjuvant therapy to prevent recurrence of tumor after surgical removal. STUDY DESIGN/MATERIALS AND METHODS: Sixteen dogs with naturally occurring hemangiopericytomas were treated with surgical removal of the tumor followed by PDT using Photochlor as the photosensitizer. Photochlor was injected intravenously at a dose of 0.3 mg/kg. Forty-eight hours later the treatment consisted of surgical removal of the tumor followed by HPPH-PDT. RESULTS: Nine dogs (56%) had recurrence of tumor from 2 to 29 (median 9) months after treatment. These results are comparable or not as good as other forms of therapy. CONCLUSIONS: Photochlor photodynamic therapy applied after surgery appears to have no advantage over other forms of therapy in regards to preventing recurrence. Delayed wound healing and infections are problematic and make HPPH PDT an undesirable addition to surgery for the treatment of this tumor type. PMID- 11500859 TI - Effect of photodynamic therapy (PDT) on the expression of pro-apoptotic protein Bak in nasopharyngeal carcinoma (NPC). AB - BACKGROUND AND OBJECTIVE: To investigate the effect of photodynamic therapy (PDT) on expression of the pro-apoptotic gene Bak in nasopharyngeal carcinoma (NPC). STUDY DESIGN/MATERIALS AND METHODS: Apoptosis and expression of the pro-apoptotic gene Bak on the tumor tissues from both pre- and post-PDT were determined using the in situ end labeling (ISEL), standard immunohistochemistry technique and western blot, respectively, in 24 patients with either persistent or recurrent NPC after radiotherapy. RESULTS: Before PDT, apoptotic index (AI) in tumor tissue was 1.2 +/- 0.6. At 6, 12, 24 and 48 hours after PDT, AI were 6.5 +/- 3.1, 23.6 +/- 8.3, 67.2 +/- 14.2 and 89.3 +/- 8.1, respectively. PDT caused apoptosis in a time-dependent fashion. Immunohistochemical assay indicated that 75% (18/24) of the patients had an upgrade expression of Bak protein in their tumor tissues after PDT. Increases in expression of Bak from PDT were also confirmed by western blot analysis. CONCLUSIONS: PDT probably causes NPC cell apoptosis through an upregulation of the pro-apoptotic protein Bak expression. PMID- 11500860 TI - The effect of irrigation on peak temperatures in nerve root, dura, and intervertebral disc during laser-assisted foraminoplasty. AB - BACKGROUND AND OBJECTIVE: The Holmium: YAG (Ho: YAG) laser has been used for the ablation of prolapsed discs but alternative techniques are available, and this application remains controversial. It also has potential for the decompression of nerve roots within narrowed foraminae with the technique of endoscopic laser foraminoplasty. Traditional methods of decompression necessitate a major surgical procedure with potential destabilisation of the lumbar spinal segment. Nevertheless, minimally invasive techniques are attractive only if serious complications can be avoided. This study reports the peak temperatures reached in surrounding tissues with and without saline irrigation. STUDY DESIGN/MATERIALS AND METHODS: Investigation of the hypothesis was carried out in excised sheep lumbar spines. T-type thermocouples were used for the measurement of tissue temperatures during laser ablation of nerve root foraminae. The temperature was assessed in the nerve root, dura mater, and disc space. RESULTS: The Ho: YAG laser was effective in widening the foraminae by approximately 1.5 mm with a total energy of 4.60 kJ. This was statistically significant in both vertical and horizontal directions (P < 0.0003 and P < 0.00005, respectively). The mean temperature of the nerve root, dura, and disc space during the procedure was 44 +/- 3.1 degrees C, 42.8 +/- 4.7 degrees C, and 41 +/- 3.4 degrees C respectively. There were transient high peaks seen in the temperature profiles. Using saline irrigation at 27 ml/minutes these temperatures were reduced to 34.1 +/- 1.8 degrees C (P = 0.0002), 34.9 +/- 1.5 degrees C (P = 0.002), and 37.2 +/- 1.2 degrees C (P < 0.014), for nerve roots, dura, and disc space respectively. CONCLUSIONS: Laser ablation of bone and ligament for nerve root decompression using the Ho: YAG laser may offer substantial advantages, but the risk of serious complication may only be avoided if the technique is combined with saline irrigation. PMID- 11500861 TI - Cortical bone healing following laser osteotomy using 6.1 microm wavelength. AB - BACKGROUND AND OBJECTIVE: Use of laser wavelengths in the 6.1 microm (amide I) to 6.45 microm (amide II) regions and a macropulse width of 4.0 microseconds delivered by a computer-controlled delivery system have produced clean, deep cortical bone ablations with minimal collateral thermal injury and no char formation. The purpose of this study was to evaluate the healing of cortical bone following 6.1 microm wavelength laser osteotomy using a 4.0 microsecond pulse, and compare that response to the response of similar osteotomies made with a standard pneumatic surgical bone saw. STUDY DESIGN/MATERIALS AND METHODS: Sixteen mature rabbits were divided equally into 2, 4, 6 and 8-week post-surgical survival groups. A nitrogen driven sagittal bone saw and an FEL generating 6.1 microm wavelength in 4.0 microsecond macropulses of 22.5 +/- 2.5 mJ/pulse directed into a 200 microm diameter spot were used to make 6.6 mm linear cuts into rabbit tibial cortex, and the healing response over time was monitored. Bone saw cuts were made halfway through the thickness of the cortex. Laser cuts were directed by a computer-controlled delivery system, and were either partial or full thickness cortical cuts. Location of the cortical bone cuts (saw or laser, partial or full thickness cut, proximal or distal, medial right or medial left tibia) were randomly assigned. At each predetermined post-surgical time point, rabbits of the appropriate group were euthanized, and the tibias of each subject collected, processed for histologic evaluation, and analyzed by light microscopy. RESULTS: At 2 weeks post-surgery, bone saw cuts showed no evidence of a healing response, while both the partial and full laser cut sites were filled with trabecular bone and primitive bone marrow. By 4 weeks post-surgery, the bone saw cuts showed filling of the defect with trabecular bone and primitive marrow, and an intense osteonal remodeling of the original cortex adjacent to the cuts was evident. All laser cut defects were filled, reactive periosteal bone was being converted to osteons and consolidating, and secondary osteons were appearing in the original cortex. At 6 weeks following surgery, the bone saw defects were filed with a mixture of woven and lamellar bone. All laser defects were filled with lamellar osteons and woven bone, the osteons were remodeling from primary to secondary osteons. By 8 weeks following the surgery, all bone saw and laser cut specimens revealed complete healing. CONCLUSIONS: Histologic evaluation of osteotomy sites made in skeletally mature rabbit tibia using the 6.1 microm wavelength, 4.0 microsecond macropulse FEL, delivered at 6 Hz at the osteotomy site, reveals a healing response which is at least as good as the healing of bone saw osteotomies, and appears to proceed at a faster rate during the first 2-4 weeks following surgery. PMID- 11500862 TI - In-vivo response to free electron laser incision of the rabbit cornea. AB - BACKGROUND AND OBJECTIVE: We analyzed the in vivo ocular response to corneal incisions made by Medical Free Electron Laser (MFEL) as a function of scan rate and incision depth. Additionally, we compared biomicroscopy, optical coherence tomography (OCT), and light microscopy as ocular response diagnostic tools. STUDY DESIGN/MATERIALS AND METHODS: Rabbit corneas were incised with pulsed MFEL radiation at 2.94 microm wavelength, scalpel incisions or focal cautery treatment were used as controls. The MFEL beam scan rate ranged from 0.2 to 1.0 mm/second. Ocular effects were monitored for 2 hours postoperatively using OCT and slit lamp examination. Ocular tissue was fixed for light microscopic examination. RESULTS: Anterior chamber fibrin formation correlated with MFEL incision depth. Slower scan rates resulted in deeper incisions and greater fibrin formation. OCT was better than slit lamp biomicroscopy at identifying fibrin attachments. OCT and light microscopy proved to be excellent companion technologies. CONCLUSIONS: Deep corneal incisions in the rabbit produced by the MFEL resulted in fibrin formation in the anterior chamber. PMID- 11500863 TI - Optimal solder and power density for diode laser tissue soldering (LTS). AB - BACKGROUND AND OBJECTIVE: Laser tissue soldering (LTS) using albumin and indocyanine green dye (ICG) is an effective technique utilized in various reconstructive surgical procedures. The purpose of this study was to describe in vivo and in vitro temperature profiles of an albumin-based solder while varying ICG concentration and laser power density (PD), and to describe immediate and short-term tensile strength measurements and histology of tissue with variable ICG concentrations and PD. STUDY DESIGN/MATERIALS AND METHODS: ICG ranged from 0.31 to 20 mg/mL while PD ranged from 3.2 to 63.7 W/cm(2). Direct solder temperature measurements were obtained at 5-second intervals during laser activation. Differential temperature measurements were determined within the dermis of rat skin and the overlying solder. Eighteen rats were subjected to 2.0 cm incisions (n = 113) created on the dorsal skin followed by closure with LTS at varying PD and ICG concentrations. ICG concentrations included 0.31, 2.5, and 20 mg/mL, while PD ranged from 8.0 to 63.7 W/cm(2). Tensile strength (TS) profiles were measured immediately and 10 days post-operatively. Histological examination was performed at the time of sacrifice. RESULTS: Temperature profiles of the ICG/albumin solder differed significantly only at the highest concentration of ICG (20 mg/mL), but showed statistically significant variability at different laser PD. Using solder color changes as an endpoint of LTS, average peak solder temperature ranged from 69 degrees C at a PD of 8.0 W/cm(2), 105 degrees -120 degrees C at PD 15.9-31.8 W/cm(2), and > 200 degrees C at PD > or = 47.7 W/cm(2). Peak intradermal temperatures remained below 50 degrees C at all PDs. Varying ICG concentration only had an effect on the immediate TS of wounds at the lowest power densities. Increasing PD resulted in statistically significant increases in immediate TS up to a PD of 23.9 W/cm(2) at an ICG concentrations of 0.31 and up to a PD of 15.9 W/cm(2) at a concentration of 2.5 mg/mL. Statistically insignificant decreases in 10-day would strength resulted from higher PD. Power densities > or = 23.9 W/cm(2) showed significant thermal injury upon histologic examination. CONCLUSIONS: Power density, not ICG concentration, is the primary determinant of solder and tissue temperature during LTS. Effective and reproducible laser tissue soldering may be achieved primarily by power density control when using diode laser and ICG-based albumin solder. Alterations in PD show the most direct and predictable effects on the healing properties of skin closed by LTS. Optimal laser wound closure occurs with an ICG of 2.5 mg/mL and at a PD between 15.9 and 23.9 W/cm(2). PMID- 11500864 TI - Comparison of argon-laser welded and sutured repair of inferior vena cava in a canine model. AB - BACKGROUND AND OBJECTIVE: Advantages of laser-welded microarterial anastomoses have been reported. However, whether laser repair of large veins is advantageous is not yet known. Argon-laser welding of inferior vena cava was therefore compared with conventional-sutured repair. STUDY DESIGN/MATERIALS AND METHODS: Twenty-four adult mongrel dogs were used. Twelve inferior vena cavas were repaired using an argon laser and the rest using continuous sutures. Specimens were removed at 2, 8, 16, and 24 weeks after repair. Technical characteristics, cavographical findings, and results of histopathological study using light and transmission electron microscopy were compared. RESULTS: Procedure time did not significantly differ between the two groups. The sutured repair sites were narrower than those repaired by laser immediately after and at 2 and 24 weeks. Histologically, laser welding resulted from denaturation of collagen and mild healing occurred simultaneously with the disappearance of denatured collagen. By contrast, there was strong fibrotic reaction at sutured repair sites. CONCLUSIONS: These findings suggest that argon-laser repair of large veins is superior to continuous suturing. PMID- 11500865 TI - Laser resistance of expandable metal stents in interventional bronchoscopy: an experimental evaluation. AB - BACKGROUND AND OBJECTIVE: Nd-YAG non-contact laser therapy is used to eliminate recurrent tumor tissue within airway stents. This study was performed to determine maximum exposure time until the stent wire is destroyed as well as exposure time necessary to deliberately fracture the wire mesh. STUDY DESIGN/MATERIALS AND METHODS: Strecker stents, Wallstents, and Nitinol Accuflex stents were implanted into pig bronchi. Then Nd-YAG laser light was directed to the stent wires at different power levels. Portions of the stent surrounded by bronchial tissue as well as bare wires were investigated. RESULTS: The Strecker stent and the Wallstent revealed a considerably higher laser resistance than did the Nitinol Accuflex stent. At 15 W, minimum exposure time until wire destruction was 4.6 and 6.3 s for the Strecker stent and the Wallstent respectively. In contrast, the Nitinol Accuflex stent tolerated only 1.7 s. CONCLUSIONS: Laser therapy with bronchial wire stents can be carried out at low power levels. It should, however, be avoided in the case of Nitinol Accuflex stents due to their low heat tolerance. PMID- 11500866 TI - Laser enhancement in hepatic regeneration for partially hepatectomized rats. AB - BACKGROUND AND OBJECTIVE: The bio-stimulation effect of laser has been observed in many areas of Medicine. However, there are a few works which investigate its use for liver regeneration. Most of their results were inconclusive due to the use of high power lasers. This work was carried out to investigate the bio stimulation effect of laser in liver regeneration using low power lasers. STUDY DESIGN/MATERIALS AND METHODS: We used Wistar male rats, which were irradiated with laser light (wavelength 590 nm and intensity of 50 mW/cm(2)) for 5 minutes after 70% hepatectomy. The respiratory mitochondrial activity, the serum level of aminotransferase and the PCNA were measured. RESULTS: Our results show a dramatic increase in the mitochondrial activity for the laser treated group at 24 hours after the hepatectomy. CONCLUSIONS: We conclude that the laser promotes a bio stimulation effect on the early stages of liver regeneration without any detectable damage of the cells. PMID- 11500867 TI - "Hot" KTP-laser treatment of facial angiofibromata. AB - BACKGROUND AND OBJECTIVE: Treatment of facial angiofibromata (AF) relied largely upon cutaneous resurfacing. While effective, resurfacing affects large areas with attendant risks of dyspigmentation, infection, and scarring. We investigated the pulsed KTP (532 nm) laser energy for its high absorption by melanin and hemoglobin as a photothermal destructive method for treating AF. STUDY DESIGN/MATERIALS AND METHODS: In five patients (Fitzpatrick phototypes II-;VI), AF were treated with the KTP laser (10 ms, 20 J/cm(2), 2 mm beam) using stacked pulses (2-3.3 Hz) or passes. No cooling device was employed. Each pulse evoked puffs of steam and caused progressive flattening of AF. Normal intervening skin was strictly avoided. Patients underwent one to five sessions in which as many as 100 lesions were treated. RESULTS: Individual lesions responded with complete flattening in one or two treatments. While this effect has persisted for 18-;24 months, slow recrudescence is expected. Transient hypopigmentation and hyperpigmentation were localized to treated skin. No scarring, infection, or other adverse events were observed. Patient satisfaction with this method was high due to rapid healing time ( < 10 days), minimal pain, ease of wound care, and efficacy. CONCLUSIONS: "Hot" KTP laser is an effective and safe method of treatment for facial AF. Limiting treatment only to lesional skin allowed rapid healing and very limited adverse effects despite the increased non-specific thermal damage caused by high fluence, long pulse duration, and an absence of superficial tissue cooling. PMID- 11500868 TI - Case report: holmium laser resection and lasertripsy for intravesical ureterocele with calculus. AB - The holmium laser has established an important role in urology, particularly in the treatment of benign prostatic hyperplasia (BPH) and urolithiasis. Its light (wavelength 2140 nm) has a high coefficient of absorption in water, such that 95% of the laser energy is absorbed in 0.5 mm of water. Thus, the majority of the laser energy is contained superficially, imparting the holmium laser with excellent cutting and tissue ablation properties. It provides sharp incision with simultaneous hemostasis. Perhaps the most significant benefit of the holmium laser is its versatility when delivered through rigid or flexible endoscopes using quartz fibers of various sizes. We present the use of this laser in the ablation of an intravesical ureterocele with calculus. PMID- 11500869 TI - Excimer laser (308 nm) based transmyocardial laser revascularization: effects of the lasing parameters on myocardial histology. AB - BACKGROUND AND OBJECTIVE: The effect of the excimer laser (308 nm) parameters on transmyocardial revascularization (TMR) channels is not well defined. This study investigates the influence of the pulse repetition rate, the size of the delivery catheter and its advancement speed on the morphology of TMR channels in vivo. STUDY DESIGN/MATERIALS AND METHODS: Myocardial ablation was performed in a porcine model (N = 27) using multifiber catheters of 1.0 and 1.4 mm in diameter. The catheters were advanced into the myocardium at different speeds (1.27 and 2.54 mm/sec) while ablating at various repetition rates (10-80 Hz). The radiant exposure was kept at 35 mJ/mm(2) throughout the experiments. The channel histology was quantified by digital microscopy. RESULTS: The channel cross sectional area and the extent of the thermal damage decrease as the catheter advancement speed exceeds the ablation speed and vice versa. Within the parameters tested, advancement speed of about 1.3 mm/sec and pulse repetition rates of 40 Hz produce channels of size comparable to the catheter's diameter with moderate thermomechanical damage. CONCLUSIONS: The repetition rate, catheter size, and catheter advancement speed are closely intertwined and crucial to the histological outcome of excimer laser based TMR. PMID- 11500871 TI - Real and Hypothetical Intermediate-Valence Ag(II)/Ag(III) and Ag(II)/Ag(I) Fluoride Systems as Potential Superconductors. AB - With the aim of gauging their potential as conducting or superconducting materials, we examine the crystal structures and magnetic properties of the roughly one hundred binary, ternary, and quaternary Ag(II) and Ag(III) fluorides in the solid state reported up to date. The Ag(II) cation appears in these species usually in a distorted octahedral environment, either in an [AgF](+) infinite chain or as [AgF(2)] sheets. Sometimes one finds discrete square-planar [AgF(4)](2-) ions. The Ag(III) cation occurs usually in the form of isolated square-planar [AgF(4)](-) ions. Systems containing Ag(III) (d(8)) centers are typically diamagnetic. On the other hand, the rich spectrum of Ag(II) (d(9)) environments in binary and ternary fluorides leads to most diverse magnetic properties, ranging from paramagnetism, through temperature-independent paramagnetism (characteristic for half-filled band and metallic behavior) and antiferromagnetism, to weak ferromagnetism. Ag(II) and Ag(III) have the same d electron count as Cu(II) (d(9)) and Cu(III) (d(8)), respectively. F(-) and O(2-) ions are isoelectronic, closed-shell (s(2)p(6)) species; both are weak-field ligands. Led by these similarities, and by some experimental evidence, we examine analogies between the superconducting cuprates (Cu(II)/Cu(III)-O(2-) and Cu(II)/Cu(I)-O(2-) systems) and the formally mixed-valence Ag(II)/Ag(III)-F(-) and Ag(II)/Ag(I)-F(-) phases. For this purpose we perform electronic-structure computations for a number of structurally characterized binary and ternary Ag(I), Ag(II), and Ag(III) fluorides and compare the results with similar calculations for oxocuprate superconductors. Electronic levels in the vicinity of the Fermi level (x(2)-y(2) or z(2)) have usually strongly mixed Ag(d)/F(p) character and are Ag-F antibonding, thus providing the potential of efficient vibronic coupling (typical for d(9) systems with substantially covalent bonds). According to our computations this is the result not only of a coincidence in orbital energies; surprisingly the Ag-F bonding is substantially covalent in Ag(II) and Ag(III) fluorides. The electron density of state at the Fermi level (DOS(F)) for silver fluoride materials and frequencies of the metal-ligand stretching modes have values close to those for copper oxides. The above features suggest that properly hole- or electron-doped Ag(II) fluorides might be good BCS-type superconductors. We analyze a comproportionation/disproportionation equilibrium in the hole-doped Ag(II) fluorides, and the possible appearance of holes in the F(p) band. It seems that there is a chance of generating an Ag(III)-F(-)/Ag(II)-F(0) "ionic/covalent" curve crossing in the hole-doped Ag(II)-F(-) fluorides, significantly increasing vibronic coupling. PMID- 11500872 TI - Dioxygen Activation and Methane Hydroxylation by Soluble Methane Monooxygenase: A Tale of Two Irons and Three Proteins A list of abbreviations can be found in Section 7. AB - Methanotrophic bacteria are capable of using methane as their sole source of carbon and energy. The first step in methane metabolism, the oxidation of methane to methanol, is catalyzed by a fascinating enzyme system called methane monooxygenase (MMO). The selective oxidation of the very stable C-H bond in methane under ambient conditions is a remarkable feat that has not yet been repeated by synthetic catalysts and has attracted considerable scientific and commercial interest. The best studied MMO is a complex enzyme system that consists of three soluble protein components, all of which are required for efficient catalysis. Dioxygen activation and subsequent methane hydroxylation are catalyzed by a hydroxylase enzyme that contains a non-heme diiron site. A reductase protein accepts electrons from NADH and transfers them to the hydroxylase where they are used for the reductive activation of O(2). The third protein component couples electron and dioxygen consumption with methane oxidation. In this review we examine different aspects of catalysis by the MMO proteins, including the mechanisms of dioxygen activation at the diiron site and substrate hydroxylation by the activated oxygen species. We also discuss the role of complex formation between the different protein components in regulating various aspects of catalysis. PMID- 11500873 TI - The Structure of PMID- 11500874 TI - IBX-New Reactions with an Old Reagent. PMID- 11500875 TI - Catalytic Use of a Boron Source for Boron Enolate Mediated Stereoselective Aldol Reactions in Water This work was partially supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan. Y. M. thanks the JSPS fellowship for Japanese Junior Scientists. PMID- 11500876 TI - CH/pi Attraction: The Origin of Enantioselectivity in Transfer Hydrogenation of Aromatic Carbonyl Compounds Catalyzed by Chiral eta(6)-Arene-Ruthenium(II) Complexes This work was financially supported by a Grant-in-Aid for Scientific Research (No. 07CE2004) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan. PMID- 11500877 TI - Architecture of Polymeric Superstructures: Self-Color Tone Films Constructed by Mesoscopically Ordered Cubic Lattices. PMID- 11500878 TI - Incorporation of Peptide Isosteres into Enantioselective Peptide-Based Catalysts as Mechanistic Probes This research is supported by the U.S. National Science Foundation (CHE-9874963). We are also grateful to the U.S. NIH (GM-57595), DuPont, Eli Lilly, Glaxo-Wellcome, and Merck for research support. S.J.M. is a Fellow of the Alfred P. Sloan Foundation, a Cottrell Scholar of Research Corporation, and a Camille Dreyfus Teacher-Scholar. PMID- 11500879 TI - Ion-Pair Recognition by Nucleoside Self-Assembly: Guanosine Hexadecamers Bind Cations and Anions This research is sponsored by the Separations and Analysis program of the U.S. Department of Energy. J.D. thanks the Dreyfus Foundation for a Teacher-Scholar Award. We thank LaTarsha Riddick for help with experiments. PMID- 11500880 TI - Nickel(II) Phosphate VSB-5: A Magnetic Nanoporous Hydrogenation Catalyst with 24 Ring Tunnels A.K.C. thanks the Fondation de l'Ecole Normale Superieure and the Region de l'Ile de France for a Chaire Internationale de Recherche, Blaise Pascal. We also thank the CNRS for financial support and for providing a Poste Rouge for Q.G. and a PICS to the two groups for cooperation. The authors are indebted to D. S. Kim for his support with pore size analysis. We thank the Korean Ministry of Science and Technology (Key Research Program, KK-0005-F0) for supporting this work, and the Korea Science and Engineering Foundation (KOSEF) Fellowship for J.S.C. is gratefully acknowledged. J.S.C. was partially supported by the U.S. Department of Energy under grant DE-FG03-96ER14672. P.M.F. was supported by the National Science Foundation under the MRSEC Program (NSF-DMR-96 32716). PMID- 11500881 TI - Spherical Aromaticity of Inorganic Cage Molecules This work was supported by the Alexander von Humboldt Stiftung (Z. Chen), the Centre National de la Recherche Scientifique (H. Jiao), and the Fonds der Chemischen Industrie. We thank Prof. Dr. T. F. Fassler for his helpful discussion and reprints. PMID- 11500882 TI - Time- and Space-Resolved Luminescence of a Photonic Dye-Zeolite Antenna This work is part of the Swiss National Science Foundation Project NFP 47 (4047-057481). PMID- 11500883 TI - A Fully Characterized Complex Ion with Unreduced TCNQ as Fourfold Bridging Ligand: PMID- 11500884 TI - Targeting Molecular Recognition: Exploring the Dual Role of Functional Pseudoprolines in the Design of SH3 Ligands This work was supported by the Swiss National Science Foundation. PMID- 11500885 TI - Designed Molecules for Self-Assembly: The Controlled Formation of Two Chiral Self Assembled Polynuclear Species with Predetermined Configuration This work was financially supported by the Swiss National Science Foundation. We thank F. Nydegger for ESI-MS measurements and F. Fehr for conducting NOE experiments. Prof. C. W. Schlapfer and Prof. C. A. Daul are thanked for fruitful discussions. PMID- 11500886 TI - By Overexpression in the Yeast Pichia pastoris to Enhanced Enantioselectivity: New Aspects in the Application of Pig Liver Esterase We thank the Konrad-Adenauer foundation (St. Augustin, Germany) for a stipend to A.M., Prof. R. D. Schmid (Institute of Technical Biochemistry, Stuttgart University) for his support and discussions, and A. Gollin for the synthesis of the acetates. PMID- 11500887 TI - Hydrolysis/Polycondensation in the Solid State: Access to Crystalline Silica Based Hybrid Materials The authors are grateful to Dr. C. Dumas from the LBS laboratory at the Universite de Montpellier I for the two-dimensional X-ray analysis. PMID- 11500888 TI - A More Efficient Catalyst for the Carbonylation of Chloroarenes Palladium Catalyzed Reactions for Fine Chemical Synthesis, Part 19. This project was suported by the Solvias AG, Basel (Switzerland), and from the Bildungsministerium des Landes Mecklenburg-Vorpommern. We thank Dres. H.-U. Blaser, M. Studer, A. Schnyder, and F. Spindler (Solvias AG) for helpful contributions, M. Heyken (IfOK) for technical support, and Dr. C. Fischer, K. Kortus, and S. Buchholz (IfOK) for analytical work. Part 18: ref. PMID- 11500889 TI - Synthesis, Structure, and Reactivity of a Diphosphadiferratetrahedrane with a Fe Fe Double Bond This work was supported by the Fonds der Chemischen Industrie and by the Graduiertenkolleg "Phosphorchemie als Bindeglied verschiedener chemischer Disziplinen". PMID- 11500890 TI - Evidence for sigma Dimerization During Anodic Redox Switching of 1,3,5 Tripyrrolidinobenzene: A New Molecular Switch This work was supported by the DFG, the VW Foundation, and the Fonds der Chemischen Industrie. PMID- 11500891 TI - [] PMID- 11500892 TI - Double Photoionization of Dimethylaminobenzonitrile in Solution: A Three-Quantum Process with Intervening Chemical Step This work was supported by the Volkswagenstiftung. PMID- 11500893 TI - How Does Ethene Inactivate Cytochrome P450 En Route to Its Epoxidation? A Density Functional Study The research is supported in part by the ISF and in part by the Ministry of Science, Culture, and Sport. F.O. acknowledges the European Union for a Marie Curie Fellowship. PMID- 11500894 TI - Chameleon States: High-Valent Metal-Oxo Species of Cytochrome P450 and Its Ruthenium Analogue The research in HU was sponsored by the Binational German Israeli Foundation (GIF) and by the Israeli Ministry of Science, Culture and Sport. Partial support by the US National Science Foundation (CHE-9814301) to J.T.G. is acknowledged. F.O. thanks the EU for a Marie Curie Fellowship. PMID- 11500895 TI - The First Organically Templated Open-Framework Niobium Silicate and Germanate Phases: Low-Temperature Hydrothermal Syntheses of PMID- 11500896 TI - A Lewis Acid Catalyst Anchored on Silica Grafted with Quaternary Alkylammonium Chloride Moieties T.M.J. is grateful to the Blechner fund for a postdoctoral fellowship. PMID- 11500897 TI - Molecular Models of Titania-Silica Systems and a Late Transition Metal Complex Grafted Thereon Financial support from the Direccion General de Ensenanza Superior e Investigacion, Spain (Grant. No. D.G.E.S. PB98-0159-C02-01) is gratefully acknowledged. We thank Prof. Dr. J. L. G. Fierro, Instituto de Catalisis y Petroleoquimica, for XPS measurements. PMID- 11500898 TI - Methylenation of Aldehydes: Transition Metal Catalyzed Formation of Salt-Free Phosphorus Ylides We thank the Charette group (Universite de Montreal) for generously sharing their chemicals and equipment. This research was supported by NSERC (Canada), F.C.A.R (Quebec), the Foundation for Innovation, and the Universite de Montreal. PMID- 11500899 TI - Carbanions Substituted by Transition Metals: Synthesis, Structure, and Configurational Restrictions of a Lithium Titanium Phosphonate. PMID- 11500900 TI - Chemical Involvement of Solvent Water Molecules in Elementary Steps of the Fenton Oxidation Reaction We gratefully acknowledge the helpful discussions with Michiel Gribnau (Unilever-Vlaardingen) and we thank the Netherlands Organization for Scientific Research (NWO) for support through the PPM-CMS program and the NCF for providing computer time. PMID- 11500901 TI - Insights into the Aryl-Aryl Exchange between Palladium and Phosphane Ligands in Pd(II) Complexes: Preparation of Phthalocyanine-Containing Phosphonium Salts This work was supported by CICYT (Spain) through grant MAT-99-0180. We are indebted to Mr. G. Martin Fuchs, Laboratoire de Photonique Quantique et Moleculaire, Ecole Normale Superieure de Cachan, France, for the NLO measurements. PMID- 11500902 TI - Synthesis of Tri- and Tetracoordinate Phosphorus Compounds Containing a PCF(3) Group by Nucleophilic Trifluoromethylation of the Corresponding PF Compounds. PMID- 11500903 TI - The Thioglycoside and Glycosyl Phosphite of 5-Azido Sialic Acid: Excellent Donors for the alpha-Glycosylation of Primary Hydroxy Groups This research was supported by Academia Sinica (Taipei) and the NIH (USA). PMID- 11500904 TI - Enzyme-Activated Gd(3+) Magnetic Resonance Imaging Contrast Agents with a Prominent Receptor-Induced Magnetization Enhancement We thank Dr. Shrikumar Nair for helpful discussions. PMID- 11500905 TI - Butane-2,3-Diacetal-Desymmetrized Glycolic Acid-A New Building Block for the Stereoselective Synthesis of Enantiopure alpha-Hydroxy Acids We thank the EU (Marie Curie Fellowship to E.D.), the EPSRC (to D.J.D.), the Novartis Research Fellowship (to S.V.L.), and Pfizer Gobal Research and Development, Groton, USA, for financial support. PMID- 11500906 TI - Directed Assembly of Periodic Materials from Protein and Oligonucleotide-Modified Nanoparticle Building Blocks C.A.M. acknowledges DARPA, NSF, ARO, and NIH for support of this research. R.L.L. acknowledges the NIH. The DND-CAT Synchrotron Research Center is supported by E.I. Dupont de Nemours & Co., The Dow Chemical Company, the U.S. National Science Foundation through Grant DMR-9304725, and the State of Illinois through the Department of Commerce and the Board of Higher Education Grant IBHE HECA NWU 96. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Energy Research under Contract No. W-31-102-Eng-38. PMID- 11500907 TI - Ruthenium-Catalyzed One-Step Transformation of Propargylic Alcohols into Alkylidene Cyclobutenes: X-ray Characterization of an Ru(eta(3)-cyclobutenyl) Intermediate This work was supported by the CNRS and the Ministere de la Recherche. The authors are grateful to the latter for a PhD grant to J. LP. The authors wish to thank the European Cost Program D17/003/00 and the Region Bretagne for financial support. They are also grateful to A. Bondon for helpful assistance in NMR analysis. PMID- 11500908 TI - Tandem Intramolecular Alkyne Silylformylation-Allylsilylation: A Case of Remote 1,5-Asymmetric Induction Financial support was provided by the National Institutes of Health (National Institute of General Medical Sciences, GM58133). We are grateful to Bristol-Myers Squibb for generous financial support in the form of an Unrestricted Grant in Synthetic Organic Chemistry to J.L.L. We thank Merck Research Laboratories and DuPont Pharmaceuticals for generous financial support. PMID- 11500909 TI - Living Polymerization of Ethylene with a Titanium Complex Containing Two Phenoxy Imine Chelate Ligands. PMID- 11500910 TI - The First In(OTf)(3)-Catalyzed Conversion of Kinetically Formed Homoallylic Alcohols into the Thermodynamically Preferred Regioisomers: Application to the Synthesis of 22alpha-Sterols This research was supported by the National University of Singapore. PMID- 11500911 TI - Synthesis, Structure, and Properties of a Novel Heterooctametallic Complex Containing a Cyclic Ru(4)Ni(4) Core This work was supported by a grant (Grant No. SP/S1/F33/98) from the Department of Science and Technology, New Delhi, India and Hong Kong Research Grants Council Earmarked Grant CUHK 4022/98P. PMID- 11500912 TI - Selective Photoproduction of O(2) from the Mn(4)O(4) Cubane Core: A Structural and Functional Model for the Photosynthetic Water-Oxidizing Complex This research was supported by grants from the US National Institutes of Health (GM39932) and the National Science Foundation (CHE96-19190). Fellowship support was provided by the Japan Society for the Promotion of Science (M.Y.) and the Netherlands NOW Scholarship (P.B.). PMID- 11500913 TI - Expression and function of CCAAT/enhancer binding proteinbeta (C/EBPbeta) LAP and LIP isoforms in mouse mammary gland, tumors and cultured mammary epithelial cells. AB - CCAAT/Enhancer binding proteins (C/EBPs) play important roles in the regulation of cell growth and differentiation. This study investigated the expression and function of C/EBPbeta isoforms in the mouse mammary gland, mammary tumors, and a nontransformed mouse mammary epithelial cell line (HC11). C/EBPbeta mRNA levels are 2-5-fold higher in mouse mammary tumors derived from MMTV/c-neu transgenic mice compared with lactating and involuting mouse mammary gland. The "full length" 38 kd C/EBPbeta LAP ("Liver-enriched Activator Protein") isoform is the predominant C/EBPbeta protein isoform in mammary tumor whole cell lysates, however, the truncated 20 kd C/EBPbeta LIP ("Liver-enriched Inhibitory Protein") isoform is also present at detectable levels (mean LAP:LIP ratio 5.3:1). The mammary tumor C/EBPbeta LAP:LIP ratio decreases 70% (from 5.3:1 to 1.6:1) when lysate preparation is switched from a rapid whole cell lysis protocol to a multistep nuclear/cytoplasmic fractionation protocol. In contrast to mammary tumors, only the C/EBPbeta LAP isoform is detectable in the mammary gland whole cell and nuclear lysates; the truncated "LIP" isoform is undetectable regardless of isolation protocol. Ectopic over expression of C/EBPbeta LIP or C/EBPbeta LAP did not alter HC11 growth rates. However, C/EBPbeta LIP over expressing HC11 cells (LAP:LIP ratio of approximately 1:1) exhibited a consistent 2-4 h delay in G(0)/S phase transition. C/EBPbeta LIP overexpressing HC11 cells did not express beta-casein mRNA (mammary epithelial cell differentiation marker) in response to lactogenic hormones. This defect in beta-casein expression was not corrected by carrying out the differentiation protocol in the presence of an artificial extracellular matrix. These results demonstrate that the "full-length" C/EBPbeta LAP isoform is the predominant C/EBPbeta protein isoform expressed in mouse mammary gland in vivo and mouse mammary epithelial cell cultures in vitro. C/EBPbeta LIP detected in mammary tumor lysates may result from in vivo production or ex vivo isolation-induced proteolysis of C/EBPbeta LAP. Ectopic overexpression of C/EBPbeta LIP (LAP:LIP ratio of approximately 1:1) inhibits mammary epithelial cell differentiation (beta-casein expression). PMID- 11500914 TI - Extracellular matrix regulates alpha s1-casein gene expression in rabbit primary mammary cells and CCAAT enhancer binding protein (C/EBP) binding activity. AB - Previous studies have shown that both the signal transducer and activator of transcription 5 (STAT5) and the CCAAT enhancer binding proteins (C/EBPs) are involved in the regulation of casein gene expression by mammary epithelial cells. Prolactin (Prl) activation of STAT5 is necessary for casein gene expression. The extracellular matrix (ECM) regulates also casein gene expression. Here, we have investigated whether ECM regulates C/EBPs activity in primary rabbit mammary epithelial cells. Isolated primary mammary cells were cultured on plastic or on floating collagen I gel. Prolactin induced alphas 1-casein gene expression when cells were cultured on collagen but not on plastic. It is noteworthy that activated STAT5 was detected in both culture conditions. Several STAT5 isoforms (STAT5a, STAT5b, and other STAT5 related isoforms, some with lower molecular weight than the full-length STAT5a and STAT5b) were detected under the different culture conditions. However, their presence was not related to the expression of alphas 1-casein gene. The binding of nuclear factors to a C/EBP specific binding site and the protein level of C/EBPbeta differed in cells cultured on plastic or on collagen but these parameters were not modified by Prl. This suggests that C/EBP binding activity was regulated by ECM and not by Prl. Interestingly, these modifications were correlated to the expression of the alphas 1-casein gene. Hence, the activation of the alphas 1-casein gene expression depends on two independent signals, one delivered by Prl via the activation of STAT5, the other delivered by ECM via C/EBP. PMID- 11500915 TI - Green tea extracts decrease carcinogen-induced mammary tumor burden in rats and rate of breast cancer cell proliferation in culture. AB - Epidemiological evidence suggests tea (Camellia sinensis L.) has chemopreventive effects against various tumors. Green tea contains many polyphenols, including epigallocatechin-3 gallate (EGCG), which possess anti-oxidant qualities. Reduction of chemically induced mammary gland carcinogenesis by green tea in a carcinogen-induced rat model has been suggested previously, but the results reported were not statistically significant. Here we have tested the effects of green tea on mammary tumorigenesis using the 7,12-dimethylbenz(a)anthracene (DMBA) Sprague-Dawley (S-D) rat model. We report that green tea significantly increased mean latency to first tumor, and reduced tumor burden and number of invasive tumors per tumor-bearing animal; although, it did not affect tumor number in the female rats. Furthermore, we show that proliferation and/or viability of cultured Hs578T and MDA-MB-231 estrogen receptor-negative breast cancer cell lines was reduced by EGCG treatment. Similar negative effects on proliferation were observed with the DMBA-transformed D3-1 cell line. Growth inhibition of Hs578T cells correlated with induction of p27(Kip1) cyclin dependent kinase inhibitor (CKI) expression. Hs578T cells expressing elevated levels of p27(Kip1) protein due to stable ectopic expression displayed increased G1 arrest. Thus, green tea had significant chemopreventive effects on carcinogen induced mammary tumorigenesis in female S-D rats. In culture, inhibition of human breast cancer cell proliferation by EGCG was mediated in part via induction of the p27(Kip1) CKI. PMID- 11500916 TI - Caveolae and the organization of carbohydrate metabolism in vascular smooth muscle. AB - We have previously found that glycolysis and gluconeogenesis occur in separate "compartments" of the VSM cell. These compartments may result from spatial separation of glycolytic and gluconeogenic enzymes (Lloyd and Hardin [1999] Am J Physiol Cell Physiol. 277:C1250-C1262). We have also found that an intact plasma membrane is essential for compartmentation to exist (Lloyd and Hardin [2000] Am J Physiol Cell Physiol. 278:C803-C811), suggesting that glycolysis and gluconeogenesis may be associated with distinct plasma membrane microdomains. Caveolae are one such microdomain, in which proteins of related function colocalize. Thus, we hypothesized that membrane-associated glycolysis occurs in association with caveolae, while gluconeogenesis is localized to non-caveolae domains. To test this hypothesis, we disrupted caveolae in vascular smooth muscle (VSM) of pig cerebral microvessels (PCMV) with beta methyl-cyclodextrin (CD) and examined the metabolism of [2-(13)C]glucose (a glycolytic substrate) and [1 (13)C]fructose 1,6-bisphosphate (FBP, a gluconeogenic substrate in PCMV) using (13)C nuclear magnetic resonance spectroscopy. Caveolar disruption reduced flux of [2-(13)C]glucose to [2-(13)C]lactate, suggesting that caveolar disruption partially disrupted the glycolytic pathway. Caveolae disruption may also have resulted in a breakdown of compartmentation, since conversion of [1-(13)C]FBP to [3-(13)C]lactate was increased by CD treatment. Alternatively, the increased [3 (13)C]lactate production may reflect changes in FBP uptake, since conversion of [1-(13)C]FBP to [3-(13)C]glucose was also elevated in CD-treated cells. Thus, a link between caveolar organization and metabolic organization may exist. PMID- 11500917 TI - Heat shock proteins of chicken lens. AB - The presence of heat shock proteins HSP-40, HSP-70, and HSc-70 in adult and embryonic chicken lenses were determined. The epithelium, cortex, and nucleus of adult chicken lens were separated and tested for the presence of heat shock proteins (hsps) by western blot, using specific antibodies for HSP-40, HSP-70, and HSc-70. Water soluble (WSF) and water insoluble fractions (WIF) of embryonic chicken lenses were isolated and tested for the presence of HSP-40, HSP-70, and HSc-70 by immunoblot. Embryonic chicken lens sections were also analyzed for the presence of heat shock proteins by immunofluorescence technique. Data obtained from these experiments revealed that HSP-40, HSP-70, and HSc-70 are present in all areas of both adult and embryonic chicken lens. Presence of hsps protein in the deep cortex and nucleus is intriguing as no detectable metabolic activities are reported in this area. However it can be proposed that hsps HSP-40, HSP-70, and HSc-70 can interact with protein of these areas and protect them from stress induced denaturation. PMID- 11500918 TI - Upregulation of the gene encoding a cytoplasmic dynein intermediate chain in senescent human cells. AB - Normal human somatic cells, unlike cancer cells, stop dividing after a limited number of cell divisions through the process termed cellular senescence or replicative senescence, which functions as a tumor-suppressive mechanism and may be related to organismal aging. By means of the cDNA subtractive hybridization, we identified eight genes upregulated during normal chromosome 3-induced cellular senescence in a human renal cell carcinoma cell line. Among them is the DNCI1 gene encoding an intermediate chain 1 of the cytoplasmic dynein, a microtubule motor that plays a role in chromosome movement and organelle transport. The DNCI1 mRNA was also upregulated during in vitro aging of primary human fibroblasts. In contrast, other components of cytoplasmic dynein showed no significant change in mRNA expression during cellular aging. Cell growth arrest by serum starvation, contact inhibition, or gamma-irradiation did not induce the DNCI1 mRNA, suggesting its specific role in cellular senescence. The DNCI1 gene is on the long arm of chromosome 7 where tumor suppressor genes and a senescence-inducing gene for a group of immortal cell lines (complementation group D) are mapped. This is the first report that links a component of molecular motor complex to cellular senescence, providing a new insight into molecular mechanisms of cellular senescence. PMID- 11500919 TI - MCF-7/VD(R): a new vitamin D resistant cell line. AB - Several in vitro and in vivo experiments have demonstrated potent cell regulatory effects of vitamin D compounds in cancer cells. Moreover, a promising phase I study with the vitamin D analogue Seocalcitol (EB 1089) in patients with advanced breast and colon cancer has already been carried out and more clinical trials evaluating the clinical effectiveness of EB 1089 in other cancer types are in progress (Mork Hansen et al. [2000a]). However, only little is known about the mechanisms underlying the actions of vitamin D or about the possible development of drug resistance in the patients. Therefore, in an attempt to gain more insight into these aspects, we have developed the MCF-7/VD(R) cell line, a stable subclone of the human MCF-7 breast cancer cell line, which is resistant to the growth inhibitory and apoptosis inducing effects of 1alpha,25(OH)(2)D(3). Despite this characteristic, receptor studies on the VDR have clearly demonstrated that the MCF-7/VD(R) cells contain fully functional VDRs, although in a lower number than seen with the parental MCF-7 cells. The regulation of the 24-hydroxylase enzyme appeared to be intact in the MCF-7/VD(R) cells and no differences with regard to growth rate and morphological appearance between the MCF-7/VD(R) cells and the parental MCF-7 cells were observed. Interestingly, however, the sensitivity of the MCF-7/VD(R) cells to the pure anti-estrogen ICI 182,780 was found to be increased. The MCF-7/VD(R) cell line shows characteristics different from those of previously described vitamin D resistant breast cancer cell lines but also some similarities. Together such vitamin D resistant cell lines therefore serve as a useful tool for studying the exact mechanism of action of vitamin D and the development of vitamin D resistance. PMID- 11500920 TI - Oxidative stress signalling in the apoptosis of Jurkat T-lymphocytes. AB - The pathways of transduction of oxidative stress signals have been studied using the Jurkat T cell model. The oxidative stress was induced by exposure of the cells to 100 microM H(2)O(2). DNA damage was detected within 15 min after commencement of treatment. DNA damage repair occurred within about 1 h in cells exposed to oxidative stress for 15 min. In continuous exposure to stress, DNA repair was slower and control levels of DNA integrity were not reached. DNA repair did not involve gene transcription. H(2)O(2) at 100 microM caused cell death by necrosis as well as by apoptosis. Both these processes were induced by 15 min exposure to the stress stimulus. However, some important differences were found between necrosis and apoptosis. Necrosis was more rapid, began within an hour of treatment and continued to increase during the full duration of the experiment. But apoptosis was seen after 4 h from treatment and was conspicuous between 6 and 20 h after the start of treatment. The necrotic phase preceded apoptosis, although these did show an overlap. In the necrotic phase, Bcl-2, Caspase 8 genes were down regulated. The 6-20 h phase characterised by a marked increase in apoptosis is accompanied by the up regulation of both Bcl-2 and Caspase genes. Expression of the Fas and p53 genes was not altered in either phase. We also analysed the levels of expression of the scavenging genes whose gene products are involved in detoxification. No modulation of the antioxidant enzymes, catalase, Cu/Zn superoxide dismutase and glutathione peroxidase was detectable. PMID- 11500921 TI - Apoptosis induced by progesterone in human ovarian cancer cell line SNU-840. AB - Progesterone has been used as an ingredient of anticancer drug for patients with ovarian carcinoma. However, the mechanism of anticancer effects by progesterone has not been understood. In this study, the effects of progesterone on ovarian cancer cells, SNU-840, were investigated. After the incubation with progesterone, the viability of the cells was evaluated by MTT assay. As a result, 45% of the cells were viable after 48 h of incubation with 100 microM progesterone. In addition, [(3)H]thymidine incorporation assay showed that the proliferation of the cells was completely inhibited by progesterone after 48 h of incubation at 100 microM concentration. Colorimetric TUNEL assay revealed the fragmentation of the chromosomal DNA, suggesting that the process of the cell death was apoptosis. The level of the p53 mRNA was determined by northern blotting assay, since many apoptosis processes are mediated by up-regulation of the p53 expression. The level of the p53 mRNA reached its maximum at 12 h and decreased after 24 h of incubation with progesterone. In conclusion, progesterone inhibits the proliferation and elicits apoptosis of SNU-840 cells. Also, it up-regulates the p53 mRNA transiently. PMID- 11500922 TI - Nerve growth factor stimulates the interaction of ZIP/p62 with atypical protein kinase C and targets endosomal localization: evidence for regulation of nerve growth factor-induced differentiation. AB - Atypical protein kinase Cs zeta and lambda/iota play a functional role in the regulation of NGF-induced differentiation and survival of pheochromocytoma, PC12 cells [Coleman and Wooten, 1994; Wooten et al., 1999]. Here we demonstrate an NGF dependent interaction of aPKC with its binding protein, ZIP/p62. Although, ZIP/p62 was not a PKC-iota substrate, the formation of a ZIP/p62-aPKC complex in PC12 cells by NGF occurred post activation of PKC-iota and was regulated by the tyrosine phosphorylation state of aPKC. Furthermore, NGF-dependent localization of ZIP/p62 was observed within vesicular structures, identified as late endosomes by colocalization with a Rab7 antibody. Both ZIP/p62 as well as PKC-iota colocalized with Rab7 upon NGF stimulation. Inhibition of the tyrosine phosphorylation state of PKC-iota did not prevent movement of ZIP/p62 to the endosomal compartment. These observations indicate that the subcellular localization of ZIP/p62 does not depend entirely upon activation of aPKC itself. Of functional importance, transfection of an antisense p62 construct into PC12 cells significantly diminished NGF-induced neurite outgrowth. Collectively, these findings demonstrate that ZIP/p62 acts as a shuttling protein involved in routing activated aPKC to an endosomal compartment and is required for mediating NGF's biological properties. PMID- 11500923 TI - Subcellular distribution of native estrogen receptor alpha and beta isoforms in rabbit uterus and ovary. AB - The association of estrogen receptors with non-nuclear/cytoplasmic compartments in target tissues has been documented. However, limited information is available on the distribution of estrogen receptor isoforms, specially with regard to the newly described beta isotype. The subcellular localization of estrogen receptor alpha and beta isoforms was investigated in rabbit uterus and ovary. Native alpha and beta subtypes were immunodetected using specific antibodies after subjecting the tissue to fractionation by differential centrifugation. The ovary expressed alpha and beta estrogen receptors in predominant association to cytosolic components. However, in the uterus, a substantial proportion of the total estrogen binding capacity and coexpression of the two isoforms was detected in mitochondria and microsomes. The mitochondrial-enriched subfraction represented an important source of 17beta-estradiol binding, where the steroid was recognized in a stereospecific and high affinity manner. The existence of mitochondrial and membrane estrogen binding sites correlated with the presence of estrogen receptor alpha but mainly with estrogen receptor beta proteins. Using macromolecular 17beta-estradiol derivatives in Ligand Blot studies, we could confirm that both alpha and beta isoforms were expressed as the major estrogen binding proteins in the uterus, while estrogen receptor alpha was clearly the dominant isoform in the ovary. Other low molecular weight estrogen receptor alpha-like proteins were found to represent an independent subpopulation of uterine binding sites, expressed to a lesser extent. This differential cellular partitioning of estrogen receptor alpha and beta forms may contribute to the known diversity of 17beta estradiol effects in target organs. Both estrogen receptor alpha and beta expression levels and cellular localization patterns among tissues, add complexity to the whole estrogen signaling system, in which membrane and mitochondrial events could also be implicated. PMID- 11500924 TI - Constitutive and heat-shock induced expression of Hsp70 mRNA during chicken testicular development and regression. AB - The constitutive and heat shock induced expression of Hsp70 mRNA was investigated in normal adult chicken testis and in adult testis after testicular regression induced by diethylstilbestrol (DES) treatment. In addition to the canonical form of Hsp70 mRNA, we have detected transcripts with an extended 5'UTR and transcripts containing, in the 5'UTR, sequences of the 18S ribosomal RNA. Hsp70 was expressed in unstressed male gonads in adult and regressed testis, being the expression much lower in regressed testis. Upon heat shock at 44 degrees C or 46 degrees C, Hsp70 was highly induced in both tissues. However, when testicular seminiferous tubules were incubated at the chicken internal temperature of 39 degrees C, no induction of Hsp70 was observed in mature testis, while the expression markedly increased in regressed testis. Induction at 39 degrees C was completely inhibited in the presence of 6 mM aspirin. Aspirin in the range 3-10 mM decreases the expression of Hsp70 in unstressed and stressed testicular cells, in striking contrast with the effect observed in other tissues as liver. These data suggest that the expression of Hsp70 is regulated in a specific manner in chicken testis and particularly in the male gonad undergoing regression. PMID- 11500925 TI - Induction of oxidative stress by homocyst(e)ine impairs endothelial function. AB - Previous studies have demonstrated a relationship between hyperhomocysteinemia and endothelial dysfunction, reduced bioavailability of nitric oxide, elastinolysis and, vascular muscle cell proliferation. In vivo decreased nitric oxide production is associated with increased matrix metalloproteinase (MMP) activity and formation of nitrotyrosine. To test the hypothesis that homocysteine neutralizes vascular endothelial nitric oxide, activates metalloproteinase, causes elastinolysis and vascular hypertrophy, we isolated aortas from normotensive Wistar rats and cultured them in medium containing homocysteine, and calf serum for 14 days. Homocysteine-mediated impairment of endothelial-dependent vasodilatation was reversed by co-incubation of homocysteine with nicotinamide (an inhibitor of peroxinitrite and nitrotyrosine), suggesting a role of homocysteine in redox-mediating endothelial dysfunction and nitrotyrosine formation. The Western blot analysis, using anti-nitrotyrosine antibody, on aortic tissue homogeneates demonstrated decreased nitrotyrosine in hyperhomocysteinemic vessels treated with nicotinamide. Zymographic analysis revealed increased elastinolytic gelatinase A and B (MMP-2, -9) in homocysteine treated vessels and the treatment with nicotinamide decreases the homocysteine induced MMP activation. Morphometric analyses revealed significant medial hypertrophic thickening (1.4 +/- 0.2-fold of control, P = 0.03) and elastin disruption in homocysteine-treated vessels as compared to control. To determine whether homocysteine causes endothelial cell injury, cross-sections of aortas were analyzed for caspase activity by incubating with Ac-YVAD-AMC (substrate for apoptotic enzyme, caspase). The endothelium of homocysteine treated vessels, and endothelial cells treated with homocysteine, showed marked labeling for caspase. The length-tension relationship of homocysteine treated aortas was shifted to the left as compared to untreated aortas, indicating reduced vascular elastic compliance in homocysteine-treated vessels. Co-incubation of homocysteine and inhibitors of MMP, tissue inhibitor of metalloproteinase-4 (TIMP-4), and caspase, YVAD-CHO, improved vascular function. The results suggest that alteration in vascular elastin/collagen ratio and activation of MMP-2 are associated with decreased NO production in hyperhomocysteinemia. PMID- 11500926 TI - Effects of zinc on cell proliferation and proteoglycan characteristics of epiphyseal chondrocytes. AB - Zinc has been postulated as an important nutritional factor involved in growth promotion; however, the cellular mechanisms involved in the effects of zinc on linear growth remain to be elucidated. This study was conducted to evaluate the effects of zinc on the proliferation rate of epiphyseal growth plate chondrocytes and on the structural characteristics of the proteoglycans synthesized by these cells. For these purposes, hypertrophic and proliferating chondrocytes were isolated from the tibiae of 1- and 5-week-old chickens, respectively. Chondrocytes were cultured under serum-free conditions and primary cultures were used. The results showed that zinc stimulated proliferation by 40-50% above the baseline in the case of proliferating chondrocytes, but it had no effect on hypertrophic chondrocytes. Zinc had neither any effects on mean charge density of proteoglycans synthesized by hypertrophic chondrocytes nor in their hydrodynamic size. In contrast, zinc induced an increase in mean charge density and a decrease of hydrodynamic size of proteoglycans synthesized by proliferating chondrocytes. In both cell types zinc had no effect on the composition and hydrodynamic size of the glycosaminoglycan chains. The increased ability of proliferating chondrocytes cultured in the presence of zinc to synthesize 3'-phosphoadenosine 5' phosphosulfate (PAPS) could be explained by the induction of enzymes participating in the sulfation pathway of proteoglycans. Therefore, the increase in mean charge density of proteoglycans observed in this study may be explained by an increase of the degree of sulfation of proteoglycan molecules. We speculate that the effect of zinc on linear growth may be explained at a cellular level by: a) an increase in proliferation rates of proliferating chondrocytes, and b) increased synthesis of highly charged proteoglycan molecules which decreases mineralization. PMID- 11500927 TI - Salvianolic acid B attenuates VCAM-1 and ICAM-1 expression in TNF-alpha-treated human aortic endothelial cells. AB - Attachment to, and migration of leukocytes into the vessel wall is an early event in atherogenesis. Expression of cell adhesion molecules by the arterial endothelium may play a major role in atherosclerosis. It has been suggested that antioxidants inhibit the expression of adhesion molecules and may thus attenuate the processes leading to atherosclerosis. In the present study, the effects of a potent water-soluble antioxidant, salvianolic acid B (Sal B), and an aqueous ethanolic extract (SME), both derived from a Chinese herb, Salvia miltiorrhiza, on the expression of endothelial-leukocyte adhesion molecules by tumor necrosis factor-alpha (TNF-alpha)-treated human aortic endothelial cells (HAECs) were investigated. When pretreated with SME (50 and 100 microg/ml), the TNF-alpha induced expression of vascular adhesion molecule-1 (VCAM-1) was notably attenuated (77.2 +/- 3.2% and 80.0 +/- 2.2%, respectively); and with Sal B (1, 2.5, 5, 10, and 20 microg/ml), 84.5 +/- 1.9%, 78.8 +/- 1.2%, 58.9 +/- 0.4%, 58.7 +/- 0.9%, and 57.4 +/- 0.3%, respectively. Dose-dependent lowering of expression of intercellular cell adhesion molecule-1 (ICAM-1) was also seen with SME or Sal B. In contrast, the expression of endothelial cell selectin (E-selectin) was not affected. SME (50 microg/ml) or Sal B (5 microg/ml) significantly reduced the binding of the human monocytic cell line, U937, to TNF-alpha-stimulated HAECs (45.7 +/- 2.5% and 55.8 +/- 1.2%, respectively). SME or Sal B significantly inhibited TNF-alpha-induced activation of nuclear factor kappa B (NF-kappaB) in HAECs (0.36- and 0.48-fold, respectively). These results demonstrate that SME and Sal B have anti-inflammatory properties and may explain their anti atherosclerotic properties. This new mechanism of action of Sal B and SME, in addition to their previously reported inhibition of LDL, may help explain their efficacy in the treatment of atherosclerosis. PMID- 11500928 TI - Immortalization of human WI38 cells is associated with differential activation of the c-myc origins. AB - To study the possible relationships between origin activities and cellular processes leading to malignancy, we used an isogenic system of human embryo lung fibroblast cells WI38 and a SV40-transformed variant, WI38 VA13 2RA (WI38(SV40)). We found that the activities of all initiation sites at the c-myc locus were approximately two-fold as high in WI38(SV40) cells as in WI38 cells. Thus, higher initiation frequency of origins at certain loci is induced with cell immortalization, one of the steps in the multi-step process leading to malignancy. We measured the activities of the four c-myc promoters P0, P1, P2, and P3 with nuclear runon assay in the two cell lines in order to detect potential individual promoter changes that may be also associated with immortalization by SV40 virus. The results show that the activities of the promoters P0, P1, and P3 did not significantly change, but the activity of the major promoter P2 in WI38(SV40) cells was about 7.5- to 8.0-fold as high as that in WI38 cells. The increased activity of promoter P2, although approximately 600 bp downstream of one of the major DNA replication initiation sites, had no preferential influence on the major sites of origin activity. Since the distribution of nascent strand abundance was not significantly altered, binding of transcription factors does not seem to facilitate the assembly of pre replication complex (pre-RC) or otherwise preferentially alter the activities of the DNA replication proteins at this major initiation site. PMID- 11500931 TI - Inhibition of nitric oxide synthase inhibitors and lipopolysaccharide induced inducible NOS and cyclooxygenase-2 gene expressions by rutin, quercetin, and quercetin pentaacetate in RAW 264.7 macrophages. AB - Several natural flavonoids have been demonstrated to perform some beneficial biological activities, however, higher-effective concentrations and poor absorptive efficacy in body of flavonoids blocked their practical applications. In the present study, we provided evidences to demonstrate that flavonoids rutin, quercetin, and its acetylated product quercetin pentaacetate were able to be used with nitric oxide synthase (NOS) inhibitors (N-nitro-L-arginine (NLA) or N-nitro L-arginine methyl ester (L-NAME)) in treatment of lipopolysaccharide (LPS) induced nitric oxide (NO) and prostaglandin E2 (PGE2) productions, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) gene expressions in a mouse macrophage cell line (RAW 264.7). The results showed that rutin, quercetin, and quercetin pentaacetate-inhibited LPS-induced NO production in a concentration dependent manner without obvious cytotoxic effect on cells by MTT assay using 3 [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide as an indicator. Decrease of NO production by flavonoids was consistent with the inhibition on LPS induced iNOS gene expression by western blotting. However, these compounds were unable to block iNOS enzyme activity by direct and indirect measurement on iNOS enzyme activity. Quercetin pentaacetate showed the obvious inhibition on LPS induced PGE2 production and COX-2 gene expression and the inhibition was not result of suppression on COX-2 enzyme activity. Previous study demonstrated that decrease of NO production by L-arginine analogs effectively stimulated LPS induced iNOS gene expression, and proposed that stimulatory effects on iNOS protein by NOS inhibitors might be harmful in treating sepsis. In this study, NLA or L-NAME treatment stimulated significantly on LPS-induced iNOS (but not COX-2) protein in RAW 264.7 cells which was inhibited by these three compounds. Quercetin pentaacetate, but not quercetin and rutin, showed the strong inhibitory activity on PGE2 production and COX-2 protein expression in NLA/LPS or L-NAME/LPS co-treated RAW 264.7 cells. These results indicated that combinatorial treatment of L-arginine analogs and flavonoid derivates, such as quercetin pentaacetate, effectively inhibited LPS-induced NO and PGE2 productions, at the same time, inhibited enhanced expressions of iNOS and COX-2 genes. PMID- 11500932 TI - Stromelysin-3 suppresses tumor cell apoptosis in a murine model. AB - Stromelysin-3 (STR-3) is a matrix metalloproteinase with a unique pattern of expression and substrate specificity. During embryogenesis and remodeling of normal adult tissues, STR-3 is produced by stromal cells in direct contact with epithelial cells undergoing regional apoptosis and selective cell survival. STR-3 is also overexpressed by interdigitating stromal cells in primary epithelial malignancies. Although STR-3 does not degrade classic extracellular matrix components, the enzyme promotes the establishment of local tumors in nude mice by as yet undefined mechanisms. STR-3 is induced when malignant epithelial cells come into contact with surrounding stromal elements; the active stromal cell derived 45 kDa enzyme is subsequently processed to a 35 kDa protein without enzymatic activity. We have generated MCF-7 transfectants expressing wild type or catalytically inactive 45 kDa STR-3 (STR-3wt and STR-3cat-) or secreted 35 kDa STR-3 (35 kDa STR-3sec) and evaluated their implantation and survival in nude mice. Tumors developed significantly more rapidly in animals receiving STR-3wt, rather than vector-only, STR-3cat- or 35 kDa STR-3sec transfectants. Most importantly, STR-3wt tumors had a significantly lower percentage of apoptotic cells than tumors derived from vector-only, STR-3cat- or 35 kDa STR-3sec transfectants. Taken together, these studies suggest that the active STR-3 enzyme may increase tumor take by suppressing tumor cell apoptosis and that 45 kDa to 35 kDa STR-3 processing limits STR-3 activity at the tumor/stromal interface. Because STR-3 is secreted as an active enzyme rather than a proform, subsequent 45 kDa to 35 kDa STR-3 processing may represent a novel mechanism for regulating enzymatic activity. PMID- 11500933 TI - p38beta MAP kinase protects rat mesangial cells from TNF-alpha-induced apoptosis. AB - p38 MAP kinases (p38) and c-Jun N-terminal protein kinases (JNK) have been associated with TNF-alpha-induced apoptosis. However, recent studies indicate that an early but brief activation of JNK and/or p38 may actually protect some cells from TNF-alpha-induced apoptosis. Whether the activation of JNK and p38 provides a pro- or anti-apoptotic signal for TNF-alpha has been controversial. In this study, we investigated the role of p38 in the regulation of TNF-alpha cytotoxicity in rat mesangial cells. Treatment of the cells with TNF-alpha alone had little effect on their viability, but they became very sensitive to apoptosis when treated with TNF-alpha in the presence of the p38 inhibitor SB 203580. These results suggested that the p38 pathway is critical for mesangial cells to survive the toxic effect of TNF-alpha. Using adenovirus-mediated gene transfer technique, we further demonstrated that p38beta, but not p38alpha, is essential to protect the cells from TNF-alpha toxicity. It has been speculated that there is a synergetic interaction between the p38 and the nuclear factor-kappaB (NF-kappaB) pathways in protecting certain cells from apoptosis. However, expression of neither p38beta nor its dominant negative mutant in mesangial cells interfered with TNF-alpha-induced translocation of NF-kappaB, the initial step of NF-kappaB activation. While it is unclear whether p38beta regulates NF-kappaB transcription activity at other steps, it is apparent that p38beta does not affect TNF-alpha induced NF-kappaB activation at the stage of nuclear translocation. PMID- 11500934 TI - Expression patterns of vitamin D receptor in human prostate. AB - UV exposure and serum levels of vitamin D have been linked in several studies with prostate cancer risk. At the cellular level, the principal action of vitamin D is mediated though vitamin D receptors (VDR). Since prostate cancer is a disease strongly associated with age, we examined the presence of VDR in normal prostate from donors of various ages to determine if the VDR expression pattern changed with age. We also compared the VDR expression in the peripheral and central zones of the prostate to determine if the expression pattern varied by location. Immunohistochemical studies were performed on paraffin-embedded tissue from cases selected by the following age decades; 10-19, 20-29, 30-39, 40-49, 50 59, and 60-69. Both the central and peripheral zones were examined for VDR expression. The intensity of VDR expression in prostate was compared with expression in different types of human tissues. Mean VDR expression was lowest in the 10-19 years of age group. The intensity of the nuclear VDR was higher though the fifth decade, and then declined in cases of ages 60-70. When multiple sections of the same donor prostate were compared, VDR expression was greater in the peripheral zone compared to the central zone. PMID- 11500935 TI - Chromatin and histones from Giardia lamblia: a new puzzle in primitive eukaryotes. AB - The three deepest eukaryote lineages in small subunit ribosomal RNA phylogenies are the amitochondriate Microsporidia, Metamonada, and Parabasalia. They are followed by either the Euglenozoa (e.g., Euglena and Trypanosoma) or the Percolozoa as the first mitochondria-containing eukaryotes. Considering the great divergence of histone proteins in protozoa we have extended our studies of histones from Trypanosomes (Trypanosoma cruzi, Crithidia fasciculata and Leishmania mexicana) to the Metamonada Giardia lamblia, since Giardia is thought to be one of the most primitive eukaryotes. In the present work, the structure of G. lamblia chromatin and the histone content of the soluble chromatin were investigated and compared with that of higher eukaryotes, represented by calf thymus. The chromatin is present as nucleosome filaments which resemble the calf thymus array in that they show a more regular arrangement than those described for Trypanosoma. SDS-polyacrylamide gel electrophoresis and protein characterization revealed that the four core histones described in Giardia are in the same range of divergence with the histones from other lower eukaryotes. In addition, G. lamblia presented an H1 histone with electrophoretic mobility resembling the H1 of higher eukaryotes, in spite of the fact that H1 has a different molecular mass in calf thymus. Giardia also presents a basic protein which was identified as an HU-like DNA-binding protein usually present in eubacteria, indicating a chimaeric composition for the DNA-binding protein set in this species. Finally, the phylogenetic analysis of selected core histone protein sequences place Giardia divergence before Trypanosoma, despite the fact that Trypanosoma branch shows an acceleration in the evolutionary rate pointing to an unusual evolutionary behavior in this lineage. PMID- 11500936 TI - Age-related decline in the osteogenic potential of human bone marrow cells cultured in three-dimensional collagen sponges. AB - Studies with human and animal culture systems indicate that a sub-population of bone marrow stromal cells has the potential to differentiate into osteoblasts. There are conflicting reports on the effects of age on human marrow-derived osteogenic cells. In this study, we used a three dimensional (3D) culture system and quantitative RT-PCR methods to test the hypothesis that the osteogenic potential of human bone marrow stromal cells decreases with age. Marrow was obtained from 39 men aged 37 to 86 years, during the course of total hip arthroplasty. Low-density mononuclear cells were seeded onto 3D collagen sponges and cultured for 3 weeks. Histological sections of sponges were stained for alkaline phosphatase activity and were scored as positive or negative. In the group < or = 50 years, 7 of 11 samples (63%) were positive, whereas only 5 of 19 (26%) of the samples in the group > or = 60 years were positive (p = 0.0504). As revealed by RT-PCR, there was no expression of alkaline phosphatase or collagen type I mRNA before culture, however there were strong signals after 3 weeks, an indication of osteoblast differentiation in vitro. We performed a quantitative, competitive RT-PCR assay with 8 samples (age range 38-80) and showed that the group < or = 50 years had 3-fold more mRNA for alkaline phosphatase than the group > or = 60 years (p = 0.021). There was a significant decrease with age (r = - 0.78, p = 0.028). These molecular and histoenzymatic data indicate that the osteogenic potential of human bone marrow cells decreases with age. PMID- 11500937 TI - Involvement of p38 MAP kinase in TGF-beta-stimulated VEGF synthesis in aortic smooth muscle cells. AB - Although it is known that transforming growth factor (TGF)-beta induces vascular endothelial growth factor (VEGF) synthesis in vascular smooth muscle cells, the underlying mechanisms are still poorly understood. In the present study, we examined whether the mitogen-activated protein (MAP) kinase superfamily is involved in TGF-beta-stimulated VEGF synthesis in aortic smooth muscle A10 cells. TGF-beta stimulated the phosphorylation of p42/p44 MAP kinase and p38 MAP kinase, but not that of SAPK (stress-activated protein kinase)/JNK (c-Jun N-terminal kinase). The VEGF synthesis induced by TGF-beta was not affected by PD98059 or U0126, specific inhibitors of the upstream kinase that activates p42/p44 MAP kinase. We confirmed that PD98059 or U0126 did actually suppress the phosphorylation of p42/p44 MAP kinase by TGF-beta in our preparations. PD169316 and SB203580, specific inhibitors of p38 MAP kinase, significantly reduced the TGF-beta-stimulated synthesis of VEGF (each in a dose-dependent manner). PD169316 or SB203580 attenuated the TGF-beta-induced phosphorylation of p38 MAP kinase. These results strongly suggest that p38 MAP kinase plays a part in the pathway by which TGF-beta stimulates the synthesis of VEGF in aortic smooth muscle cells. PMID- 11500938 TI - Tissue specific metabolism of 1alpha,25-dihydroxy-20-epi-vitamin D3 into new metabolites with significant biological activity: studies in rat osteosarcoma cells (UMR 106 and ROS 17/2.8). AB - In a recent study, we investigated the metabolism of 1alpha,25-dihydroxy-20-epi vitamin D3 (1alpha,25(OH)2-20-epi-D3), a potent synthetic vitamin D3 analog in the isolated perfused rat kidney and proposed that the enhanced biological activity of 1alpha,25(OH)2-20-epi-D3 is in part due to its metabolism into stable bioactive intermediary metabolites derived via the C-24 oxidation pathway (Siu Caldera et al. [1999] J. Steroid. Biochem. Mol. Biol. 71:111-121). It is now well established that 1alpha,25(OH)2D3 and its analogs are metabolized in target tissues not only via the C-24 oxidation pathway but also via the C-3 epimerization pathway. As the perfused rat kidney does not express the C-3 epimerization pathway, we could not identify other possible bioactive metabolites of 1alpha,25(OH)2-20-epi-D3 such as 1alpha,25(OH)2-20-epi-3-epi-D3, derived via the C-3 epimerization pathway. Therefore, we studied the metabolism of 1alpha,25(OH)2-20-epi-D3 in rat osteosarcoma cells (UMR 106) which express both the C-24 oxidation and the C-3 epimerization pathways. Our results indicate that 1alpha,25(OH)2-20-epi-D3 is metabolized in UMR 106 cells into several metabolites which included not only the previously known metabolites of the C-24 oxidation pathway but also three new metabolites which were labeled as metabolites X, Y1, and Y2. Metabolite X was unequivocally identified as 1alpha,25(OH)2-20-epi-3-epi D3. Even though definite structure identification of the metabolites, Y1 and Y2 was not achieved in our present study, we determined that the metabolite Y1 is produced from 1alpha,25(OH)2-20-epi-D3 and the metabolite Y2 is produced from 1alpha,25(OH)2-20-epi-3-epi-D3. We also noted the production of both 1alpha,25(OH)2-20-epi-3-epi-D3 and the two metabolites Y1 and Y2 in different rat osteosarcoma cells (ROS 17/2.8) which express only the C-3 epimerization pathway but not the C-24 oxidation pathway. Furthermore, we investigated the metabolism of 1alpha,25(OH)2-20-epi-D3 in the isolated perfused rat kidney in an earlier study. The results of this study indicated that the rat kidney unlike rat osteosarcoma cells did not produce either 1alpha,25(OH)2-20-epi-3-epi-D3 or the metabolites Y1 and Y2. Thus, it appears that the metabolites Y1 and Y2, like 1alpha,25(OH)2-20-epi-3-epi-D3, are produced only in specific tissues. Preliminary biological activity of each new metabolite is assessed by measuring its ability to generate VDR-mediated gene transcription. 1alpha,25(OH)2-20-epi-3 epi-D3 was found to be almost equipotent to 1alpha,25(OH)2-20-epi-D3 while the metabolites, Y1 and Y2 were found to be less active. The metabolite Y1 when compared to the metabolite Y2 has higher biological activity and its potency is almost equal to 1alpha,25(OH)2D3. In summary, we report for the first time tissue specific metabolism of 1alpha,25(OH)2-20-epi-D3 into several bioactive metabolites which are derived not only via the previously established C-24 oxidation and C-3 epimerization pathways but also via a new pathway. (c) 2001 Wiley-Liss, Inc. PMID- 11500939 TI - Regulation of the Akt/Glycogen synthase kinase-3 axis by insulin-like growth factor-II via activation of the human insulin receptor isoform-A. AB - Insulin-like growth factor II (IGF-II) plays a key role in mitogenesis during development and tumorigenesis and is believed to exert its mitogenic functions mainly through the IGF-I receptor. Recently, we identified the insulin receptor isoform A (IR(A)) as an additional high affinity receptor for IGF-II in both fetal and cancer cells. Here we investigated the mitogenic signaling of IGF-II via the Akt/Glycogen synthase kinase 3 (Gsk3) axis employing R-IR(A) cells that are IGF-I receptor null mouse embryonic fibroblasts expressing the human IR(A). IGF-II induced activation of the proto-oncogenic serine kinase Akt, reaching maximal at 5-10 min. IGF-II also caused the rapid and sustained deactivation of glycogen synthase kinase 3-beta (Gsk3beta), reaching maximal at 1-3 min, shortly preceding, therefore, maximal activation of Akt. Under our conditions, IGF-II and insulin induced 70-80% inhibition of Gsk3betaactivity. In these cells IGF-II also deactivated Gsk3alpha although less effectively than Gsk3beta. In parallel experiments, we found that IGF-II induced transient activation of extracellular signal-regulated kinases (Erk) reaching maximal at 5-10 min and decreasing thereafter. Time courses and potencies of regulation of both mitogenic pathways (Akt/Gsk3beta and Erk) by IGF-II via IR(A) were similar to those of insulin. Furthermore, IGF-II like insulin effectively stimulated cell cycle progression from the G0/G1 to the S and G2/M phases. Interestingly, AP-1-mediated gene expression, that was reported to be negatively regulated by Gsk3beta was only weakly increased after IGF-II stimulation. Our present data suggest that the coordinated activation or deactivation of Akt, Gsk3beta, and Erk may account for IGF-II mitogenic effects and support an active role for IR(A) in IGF-II action. PMID- 11500940 TI - Response of bovine endothelial cells to FGF-2 and VEGF is dependent on their site of origin: Relevance to the regulation of angiogenesis. AB - Angiogenesis, the formation of new capillary blood vessels, occurs almost exclusively in the microcirculation. This process is controlled by the interaction between factors with positive and negative regulatory activity. In this study, we have compared the effect of two well described positive regulators, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2) on bovine adrenal cortex-derived microvascular endothelial (BME) and bovine aortic endothelial (BAE) cells. The parameters we assessed included (a) cellular reorganization and lumen formation following exposure of the apical cell surface to a three-dimensional collagen gel; (b) organization of the actin cytoskeleton; (c) expression of thrombospondin-1 (TSP-1), an endogenous negative regulator of angiogenesis; and (d) extracellular proteolytic activity mediated by the plasminogen activator (PA)/plasmin system. We found that (a) collagen gel overlay induces rapid reorganization and lumen formation in BME but not BAE cells; (b) FGF-2 but not VEGF induced dramatic reorganization of actin microfilaments in BME cells, with neither cytokine affecting BAE cells; (c) FGF-2 decreased TSP-1 protein and mRNA expression in BME cells, an effect which was specific for FGF-2 and BME cells, since TSP-1 protein levels were unaffected by VEGF in BME cells, or by FGF-2 or VEGF in BAE cells; (d) FGF-2 induced urokinase type PA (uPA) in BME and BAE cells, while VEGF induced uPA and tissue-type PA in BME cells with no effect on BAE cells. Taken together, these findings reveal endothelial cell-type specific responses to FGF-2 and VEGF, and point to the greater specificity of these cytokines for endothelial cells of the microvasculature than for large vessel (aortic) endothelial cells. Furthermore, when viewed in the context of our previous observation on the synergistic interaction between VEGF and FGF-2, our present findings provide evidence for complementary mechanisms which, when acting in concert, might account for the synergistic effect. PMID- 11500941 TI - Nuclear apoptotic changes: an overview. AB - Apoptosis is a form of active cell death essential for morphogenesis, development, differentiation, and homeostasis of multicellular organisms. The activation of genetically controlled specific pathways that are highly conserved during evolution results in the characteristic morphological features of apoptosis that are mainly evident in the nucleus. These include chromatin condensation, nuclear shrinkage, and the formation of apoptotic bodies. The morphological changes are the result of molecular alterations, such as DNA and RNA cleavage, post-translational modifications of nuclear proteins, and proteolysis of several polypeptides residing in the nucleus. During the last five years our understanding of the process of apoptosis has dramatically increased. However, the mechanisms that lead to apoptotic changes in the nucleus have been only partially clarified. Here, we shall review the most recent findings that may explain why the nucleus displays these striking modifications. Moreover, we shall take into consideration the emerging evidence about apoptotic events as a trigger for the generation of autoantibodies to nuclear components. PMID- 11500942 TI - Characterization of the upstream mouse Cbfa1/Runx2 promoter. AB - Cbfa1 (or Runx2/AML-3/PEPB2alpha) is a transcriptional activator of osteoblastic differentiation. To investigate the regulation of Cbfa1 expression, we isolated and characterized a portion of the 5'-flanking region of the Cbfa1 gene containing its "bone-related" or P1 promoter and exon 1. We identified additional coding sequence in exon 1 and splice donor sites that potentially give rise to a novel Cbfa1 isoform containing an 18 amino acid insert. In addition, primer extension mapping identified in the Cbfa1 promoter a minor mRNA start site located approximately 0.8 kb 5' upstream of the ATG encoding the MASN/p57 isoform and approximately 0.4 kb upstream of the previously reported start site. A luciferase reporter construct containing 1.4 kb of the mouse Cbfa1 promoter was analyzed in Ros 17/2.8 and MC3T3-E1 osteoblast cell lines that express high levels of Cbfa1 transcripts. The activity of this construct was also examined in non-osteoblastic Cos-7 and NIH3T3 cells that do not express Cbfa1 and mesenchymal derived cell lines, including CH3T101/2, C2C12, and L929 cells, that express low levels of mature Cbfa1 transcripts. The 1.4 kb 5' flanking sequence of the Cbfa1 gene directed high levels of transcriptional activity in Ros 17/2.8 and MC3T3-E1 osteoblasts compared to non-osteoblasts Cos-7 cells, but this construct also exhibited high levels of expression in C310T1/2, L929, and C2C12 cells as well as NIH3T3 cells. In addition, Cbfa1 mRNA expression, but not the activity of the Cbfa1 promoter, was upregulated in a dose-dependent manner in pluripotent mesenchymal C2C12 by bone morphogenetic protein-2 (BMP-2). These data indicate that Cbfa1 is expressed in osteogenic as well as non-osteogenic cells and that the regulation of Cbfa1 expression is complex, possibly involving both transcriptional and post-transcriptional mechanisms. Additional studies are needed to further characterize important regulatory elements and to identify additional regions of the promoter and/or post-transcriptional events responsible for the cell-type restricted regulation of Cbfa1 expression. PMID- 11500943 TI - Iron-induced oxidative stress up-regulates calreticulin levels in intestinal epithelial (Caco-2) cells. AB - Calreticulin, a molecular chaperone involved in the folding of endoplasmic reticulum synthesized proteins, is also a shock protein induced by heat, food deprivation, and chemical stress. Mobilferrin, a cytosolic isoform of calreticulin, has been proposed to be an iron carrier for iron recently incoming into intestinal cells. To test the hypothesis that iron could affect calreticulin expression, we investigated the possible associations of calreticulin with iron metabolism. To that end, using Caco-2 cells as a model of intestinal epithelium, the mass and mRNA levels of calreticulin were evaluated as a function of the iron concentration in the culture media. Increasing the iron content in the culture from 1 to 20 microM produced an increase in calreticulin mRNA and a two-fold increase in calreticulin. Increasing iron also induced oxidative damage to proteins, as assessed by the formation of 4-hydroxy-2-nonenal adducts. Co-culture of cells with the antioxidants quercetin, dimethyltiourea and N-acetyl cysteine abolished both the iron-induced oxidative damage and the iron-induced increase in calreticulin. We postulate that the iron-induced expression of calreticulin is part of the cellular response to oxidative stress generated by iron. PMID- 11500944 TI - Molecular and cellular mechanisms of adipose secretion: comparison of leptin and angiotensinogen. AB - Besides their function of lipid storage, the adipose cells secrete a number of proteins of physiopathological importance. To get further insights into this function, which remains poorly characterized, we sought to compare the mechanisms and regulation of secretion of two individual proteins in the same cells. Leptin and angiotensinogen were chosen and assessed by radioimmunoassay and quantitative immunoblotting, respectively, in primary culture of epididymal adipose cells from young obese Zucker rats. Leptin was secreted at a steady rate of 4 ng/10(6) cells/h over 2-6 h. Despite secretion, leptin cellular content remained stable at 3 ng/10(6) cells. In contrast, the rate of angiotensinogen secretion decreased regularly from 45 arbitrary units/10(6) cells/h at 2 h, to half this value at 6 h, although cell content remained constant at 100 arbitrary units/10(6) cells. Inhibition of protein synthesis by cycloheximide depleted the cells from leptin, but not from angiotensinogen for up to 6 h. Insulin increased leptin secretion (+75%) and cell content (+70 %), without affecting angiotensinogen. Secretion of both proteins was inhibited by Golgi-disturbing agents, brefeldin A and monensin. The presence of brefeldin A led to a specific rise in leptin cell content, an effect inhibited by cycloheximide and enhanced by insulin (+80%). These data show that leptin and angiotensinogen are both secreted through Golgi-dependent pathways and that their respective intracellular pool exhibit distinct turn-over rate and insulin sensitivity. These characteristics might account for the differential response of these adipose proteins to variations in the systemic environment. PMID- 11500945 TI - Multiple role of reactive oxygen species in the arterial wall. AB - Increased oxidative stress plays an important role in vascular dysfunction and atherogenesis. Both systemic factors, such as hypercholesterolemia and hyperglycemia, and local factors, such as activation of macrophages and T cells, may contribute to oxidative stress. Oxidation of lipids in lipoproteins and cell membranes leads to functionally important modifications of proteins that affect their recognition by cell surface receptors and protein-protein interactions within the cell, including DNA binding. Oxidized LDL and extracellular oxidation modulate oxidation-sensitive signaling pathways, but it is not clear to what extent this results from receptor-mediated activation or from direct effects on the intracellular redox-balance. Extensive evidence indicates that reactive oxygen species (ROS) regulate gene expression by modulating a large number of transcription factors, including the nuclear transcription factor kappa B (NFkappaB), the peroxisome proliferator activated receptorgamma (PPARgamma), and pathways linked to apoptosis. It is also increasingly recognized that cell differentiation and proliferation, cytokine expression, and programmed cell death are determined by the interactions between oxidation-sensitive regulatory pathways previously thought to lead to distinct outcomes. Because hypercholesterolemia exerts pro-oxidant effects both intra- and extracellularly and because increased ROS formation affects vascular reactivity and atherogenesis by modulating multiple signaling pathways and transcriptional events, future investigations of its atherogenic mechanisms should place greater emphasis on the net effect of such modulation on the expression of a large spectrum of genes. One way of doing this will be by defining clusters of genes responding to hypercholesterolemic stimuli--or interventions with structurally unrelated antioxidants--in analogous ways, irrespective of what regulatory pathway they are controlled by. Microarray technologies that allow simultaneous assessment of large numbers of genes may provide a tool for this approach. PMID- 11500946 TI - Characterisation of the temporal sequence of osteoblast gene expression during estrogen-induced osteogenesis in female mice. AB - Osteoblast differentiation under in vitro conditions is associated with increased expression of non-collagenous bone proteins including osteocalcin, osteopontin, and osteonectin, the exact function of which remain poorly understood. To determine whether these proteins play an important role in the formation of mineralised bone matrix by osteoblasts in vivo, we analysed the time-course of their expression during estrogen-induced osteogenesis in female mice, and compared this with the formation of new cancellous bone. Female mice were sacrificed prior to or following treatment with 17beta-estradiol for up to 32 days (500 microg/animal/week). Total RNA was extracted from femurs, and changes in expression of genes for a range of osteoblast-derived proteins assessed by Northern blot analysis. In parallel experiments, the time course of cancellous bone formation was determined by measuring bone mineral density (BMD) of the distal femur. Estrogen led to a rapid increase in BMD, which reached significance by Day 16. This was preceded by three-fold increases in expression of alkaline phosphatase (ALP) and type I collagen (COL I) at Days 8 and 12 respectively. In contrast, osteocalcin, osteopontin, and osteonectin expression showed no change during this initial period, although modest increases were observed at later times (i.e., Days 20 and 24). Our results suggest that osteocalcin, osteopontin, and osteonectin are not involved in the initial phase of the osteogenic response to estrogen, suggesting that these non-collagenous bone proteins do not play a direct role in the formation of mineralised bone matrix by osteoblasts in vivo. PMID- 11500947 TI - Heat shock factor-4 (HSF-4a) is a repressor of HSF-1 mediated transcription. AB - Heat shock transcription factors (HSFs) regulate the expression of heat shock proteins and other molecular chaperones that are involved in cellular processes from higher order assembly to protein degradation and apoptosis. Among the human HSFs, HSF-4 is expressed as at least two splice variants. One isoform (HSF-4b) possesses a transcriptional activation domain, but this region is absent in the other isoform (HSF-4a). We have recently shown that the HSF-4a isoform represses basal transcription from heterologous promoters both in vitro and in vivo. Here we show that HSF-4a and HSF-4b have dramatically different effects on HSF-1 containing nuclear bodies, which form after heat shock. While the expression of HSF-4b colocalizes with nuclear granules, the expression of HSF-4a prevents their formation. In addition, there is a concurrent reduction of HSF-1 in the nucleus, and there is reduction in its DNA binding activity and in HSE-dependent transcription of a reporter gene. To better understand the mechanism by which HSF 4a represses transcription, we inducibly expressed HSF-4a in cells and found that HSF-4a binds to the heat shock element (HSE) during attenuation of the heat shock response. Thus HSF-4a is an active repressor of HSF-1-mediated transcription. This repressor function makes the HSF-4a isoform unique within the HSF family. PMID- 11500948 TI - Regulatory role of endogenous regucalcin in the enhancement of nuclear deoxyribonuleic acid synthesis with proliferation of cloned rat hepatoma cells (H4-II-E). AB - The role of endogenous regucalcin in the regulation of deoxyribonuleic acid (DNA) synthesis in the nuclei of the cloned rat hepatoma cells (H4-II-E) with proliferative cells was investigated. Cells were cultured for 6-96 h in a alpha minimum essential medium (alpha-MEM) containing fetal bovine serum (FBS; 1 or 10%). Cell number was significantly increased between 24 and 96 h after culture with 10% FBS; cell proliferation was markedly stimulated by culture with 10% FBS as compared with that of 1% FBS. In vitro DNA synthesis activity in the nuclei of cells was significantly elevated 6 h after culture with 10% FBS and its elevation was remarkable at 12 and 24 h after the culture. Nuclear DNA synthesis activity was significantly reduced in the presence of various protein kinase inhibitors (PD98059, staurosprine, or trifluoperazine) in the reaction mixture containing the nuclei of cells cultured for 12 and 24 h with FBS (1 and 10%). The addition of regucalcin (10(-7) and 10(-6)M) in the reaction mixture caused a significant inhibition of nuclear DNA synthesis activity. The presence of anti-regucalcin monoclonal antibody (25-100 ng/ml) in the reaction mixture containing the nuclei of cells cultured for 24 h with 10% FBS resulted in a significant increase in nuclear DNA synthesis activity. This increase was completely blocked by the addition of regucalcin (10(-6) M). The effect of anti-regucalcin antibody (100 ng/ml) in increasing nuclear DNA synthesis activity was significantly inhibited in the presence of various protein kinase inhibitors. DNA synthesis activity was significantly enhanced in the presence of anti-regucalcin antibody (100 ng/ml) in the reaction mixture containing the nuclei of cells cultured for 24 h with 10% FBS in the presence of Bay K 8644 (2.5 x 10(-6) M). Culture with Bay K 8644 did not cause a significant increase in DNA synthesis activity in the absence of anti regucalcin antibody. The present study demonstrates that endogenous regucalcin plays a suppressive role in the enhancement of nuclear DNA synthesis with proliferative cells. PMID- 11500949 TI - Mapping of intrinsic bent DNA sites in the upstream region of DNA puff BhC4-1 amplified gene. AB - We have identified bent DNA sites in the distal and proximal DNA puff BhC4-1 amplified gene promoter region of Bradysia hygida. The 2D modeling of the 3D DNA path and the ENDS ratio values calculated in this promoter region resulted in the identification of ten pronounced bent sites named BhC4B - 9 to + 1. The 1847 bp fragment (- 3697 to - 1850) in relation to the transcription start site shows multiple bending sites, BhC4B - 9 to BhC4B - 4, with periodicity approximately 300 bp. The analysis of the other identified bent region, starting at position - 957, reveals that the BhC4B + 1 bent site colocalizes with the putative BhC4-1 minimal promoter. The sequence analysis of bent site BhC4B - 4 shows a distribution of dA*dT at approximately 10 bp intervals between the middle of each tract, but intervals with more than one turn, approximately 20 bp, two helix turns, were detected in the other bent sites described here. The bent sites BhC4B - 6 and BhC4B - 4, contain two consensus sequences, with 60 bp each. The apparent molecular weight of fragments in the BhC4-1 promoter region were estimated in agarose gels and compared with the data obtained in polyacrylamide gels without and with ethidium bromide. The mobility reduction ratios (R-values) were determined, and a high R-value, 1.80, for a 1215 bp fragment in the distal promoter region and a 1.23 significant R-value for a 662 bp fragment in the proximal segment were found. To further analyze the predicted bent DNA sites in these fragments, the 2D trajectories of the 3D DNA path and other parameters, AT percentage, roll angle, ENDS ratio and DeltaG, were determined. The role of these bent sites in the BhC4-1 transcription regulation is discussed. PMID- 11500950 TI - Protein tyrosine phosphatase SHP-1 specifically recognizes C-terminal residues of its substrates via helix alpha0. AB - The catalytic domain of protein tyrosine phosphatase SHP-1 possesses distinct substrate specificity. It recognizes the P-3 to P-5 residues of its substrates via the beta5-loop-beta6 region. To study the substrate specificity further, we determined the structure of the catalytic domain of SHP-1 (C455S) complexed with a less-favorable-substrate peptide originated from SIRPalpha. The complex has disordered N-terminal peptide structure and reduced interactions between the N terminal peptide and the beta5-loop-beta6 region. This could be the basis for the lower affinity of peptide pY(427) for the catalytic domain of SHP-1. In addition, by comparing the SHP-1/less-favorable peptide complex structure with the SHP 1/substrate complex structures, we identified a novel substrate-recognition site in the catalytic domain of SHP-1. This site was formed by helix alpha0 and the alpha5-loop-alpha6 motif of SHP-1, and specifically bound residues at the P + 4 and further C-terminal positions of peptide substrates. PMID- 11500951 TI - Low-density caveolae-like membrane from Xenopus laevis oocytes is enriched in Ras. AB - Detergent-free discontinuous sucrose density gradient centrifugation was used to resolve low- and high-density membrane fractions from Xenopus laevis oocytes. Compared to high-density membrane, low-density oocyte membrane is enriched two fold in cholesterol and highly enriched in ganglioside GM1. Protein immunoblotting of membrane fractions from whole cells with polyclonal anti-human caveolin antibody detected multiple bands, including a distinctive triad with apparent molecular weights of 21, 33, and 48 kDa. To more clearly determine which of these caveolin-like protein(s) is associated with the oocyte plasma membrane, microdissection was used to separate external membrane (cortical preparations containing plasma membrane) from intracellular membrane. Cortical membrane preparations displayed a single 21-kDa caveolin-like protein in low-density membrane. Internal oocyte membrane displayed the higher molecular weight bands of 33 and 48 kDa and a lesser amount of the 21-kDa protein in low-density membrane fractions. Monoclonal anti-human Ras antibody detected a single 23-kDa immunoblot band that is enriched an average of eight-fold in low-density membrane fractions prepared from whole cells. This is the first report of caveolin-associated, low density membrane in amphibian oocytes, and is consistent with a role for caveolin and caveolae-like microdomains in oocyte signal transduction. PMID- 11500952 TI - Structural and functional studies on Troponin I and Troponin C interactions. AB - Troponin I (TnI) peptides (TnI inhibitory peptide residues 104-115, Ip; TnI regulatory peptide resides 1-30, TnI1-30), recombinant Troponin C (TnC) and Troponin I mutants were used to study the structural and functional relationship between TnI and TnC. Our results reveal that an intact central D/E helix in TnC is required to maintain the ability of TnC to release the TnI inhibition of the acto-S1-TM ATPase activity. Ca(2+)-titration of the TnC-TnI1-30 complex was monitored by circular dichroism. The results show that binding of TnI1-30 to TnC caused a three-folded increase in Ca(2+) affinity in the high affinity sites (III and IV) of TnC. Gel electrophoresis and high performance liquid chromatography (HPLC) studies demonstrate that the sequences of the N- and C-terminal regions of TnI interact in an anti-parallel fashion with the corresponding N- and C-domain of TnC. Our results also indicate that the N- and C-terminal domains of TnI which flank the TnI inhibitory region (residues 104 to 115) play a vital role in modulating the Ca(2+)- sensitive release of the TnI inhibitory region by TnC within the muscle filament. A modified schematic diagram of the TnC/TnI interaction is proposed. PMID- 11500953 TI - Stimulatory effect of endothelin-1 on Na-dependent phosphate transport and its signaling mechanism in osteoblast-like cells. AB - Endothelin-1 (ET-1) has been reported to modulate bone metabolism both in vivo and in vitro. In the present study, we investigated the effect of ET-1 on inorganic phosphate (Pi) transport in osteoblast-like cells, which is now considered to be important for the initiation of bone matrix calcification. ET-1 time- and dose-dependently stimulated Na-dependent Pi transport in mouse calvaria derived osteoblast-like MC3T3-E1 cells, and this effect was dependent on transcriptional and translational process. Kinetic analysis indicated that the change in Pi transport activity induced by ET-1 was due to alteration in the number of the Pi transporter. BQ123, a selective antagonist for ET(A) receptor, suppressed the ET-1-induced Pi transport, but BQ788, a selective antagonist for ET(B) receptor, had no effect. The inhibition of phosphoinositide hydrolysis by phospholipase C (PLC) partially attenuated the Pi transport by ET-1. Propranolol, which inhibits phosphatidic acid phosphohydrolase, also suppressed ET-1-induced Pi transport. On the contrary, indomethacin did not affect the stimulatory effect of Pi transport by ET-1. Calphostin C, a protein kinase C (PKC) inhibitor, significantly blunted the stimulatory effect of ET-1 on Pi transport. Combined effect of PMA and ET-1 on Pi transport was not additive. Pi transport induced by ET-1 was also suppressed in PKC down-regulated cells. In conclusion, the results of the present study indicate that in MC3T3-E1 osteoblast-like cells, ET-1 acting through ET receptor links to a stimulation of Pi transport via activation of PKC through both phosphoinositide and phosphatidylcholine hydrolyses. PMID- 11500954 TI - Impact of cell swelling on proliferative signal transduction in the liver. AB - Cellular swelling has emerged as an important initiator of metabolic and proliferative changes in various cells. Because of the unique regenerative capacity of the adult liver, researchers have delineated key intracellular signals that are activated following mitogens, injury, and partial hepatectomy. Although hepatocellular swelling is commonly observed following these regenerative stimuli, only recently has the relationship between cell volume increase and proliferative activity been investigated; to date, the data implicating cell volume increase with hepatocyte regeneration has been mostly indirect. Hepatocyte swelling has been demonstrated in various clinical scenarios from sepsis, hepatic resection, ischemia-reperfusion injury, glucocorticoid excess, and hyperinsulinemia. Using various in vivo and in vitro models of hepatocyte swelling, particularly hypo-osmotic stress, investigators have demonstrated changes in cellular structure: (1) cell membrane stretch, (2) cytoskeletal microtubule and microfilament reorganization, and (3) alterations in cytoskeletal-membrane complexes. Similar studies have demonstrated a causal relationship between cell volume increase and intracellular signals: (1) activation of cytoplasmic signaling cascades such as MAPKs, PI-3-K, and PKC, (2) activation of proliferative transcription factors NF-kappaB, AP-1, STATs, C/EBPs, and (3) transcription of metabolic and immediate early genes of regeneration. Through mechanotransduction, or the translation of physical changes to chemical signals, cell volume is a potent effector of these signaling events. Growing evidence demonstrates a link between these physical and chemical changes in the swelling-mediated growth in the liver. PMID- 11500955 TI - Tumor necrosis factor-alpha mediates RANK ligand stimulation of osteoclast differentiation by an autocrine mechanism. AB - Osteoblasts or bone marrow stromal cells are required as supporting cells for the in vitro differentiation of osteoclasts from their progenitor cells. Soluble receptor activator of nuclear factor-kappaB ligand (RANKL) in the presence of macrophage colony-stimulating factor (M-CSF) is capable of replacing the supporting cells in promoting osteoclastogenesis. In the present study, using Balb/c-derived cultures, osteoclast formation in both systems-osteoblast/bone marrow cell co-cultures and in RANKL-induced osteoclastogenesis-was inhibited by antibody to tumor necrosis factor-alpha (TNF-alpha), and was enhanced by the addition of this cytokine. TNF-alpha itself promoted osteoclastogenesis in the presence of M-CSF. However, even at high concentrations of TNF-alpha the efficiency of this activity was much lower than the osteoclastogenic activity of RANKL. RANKL increased the level of TNF-alpha mRNA and induced TNF-alpha release from osteoclast progenitors. Furthermore, antibody to p55 TNF-alpha receptors (TNF receptors-1) (but not to p75 TNF-alpha receptors (TNF receptors-2) inhibited effectively RANKL- (and TNF-alpha() induced osteoclastogenesis. Anti-TNF receptors-1 antibody failed to inhibit osteoclastogenesis in C57BL/6-derived cultures. Taken together, our data support the hypothesis that in Balb/c, but not in C57BL/6 (strains known to differ in inflammatory responses and cytokine modulation), TNF-alpha is an autocrine factor in osteoclasts, promoting their differentiation, and mediates, at least in part, RANKL's induction of osteoclastogenesis. PMID- 11500956 TI - Transfer of proalpha2(I) cDNA into cells of a murine model of human Osteogenesis Imperfecta restores synthesis of type I collagen comprised of alpha1(I) and alpha2(I) heterotrimers in vitro and in vivo. AB - The oim mouse is a model of human Osteogenesis Imperfecta (OI) that has deficient synthesis of proalpha2(I) chains. Cells isolated from oim mice synthesize alpha1(I) collagen homotrimers that accumulate in tissues. To explore the feasibility of gene therapy for OI, a murine proalpha2(I) cDNA was inserted into an adenovirus vector and transferred into bone marrow stromal cells isolated from oim mice femurs. The murine cDNA under the control of the cytomegalovirus early promoter was expressed by the transduced cells. Analysis of the collagens synthesized by the transduced cells demonstrated that the cells synthesized stable type I collagen comprised of alpha1(I) and alpha2(I) heterotrimers in the correct ratio of 2:1. The collagen was efficiently secreted and also the cells retained the osteogenic potential as indicated by the expression of alkaline phosphatase activity when the transduced cells were treated with recombinant human bone morphogenetic protein 2. Injection of the virus carrying the murine proalpha2(I) cDNA into oim skin demonstrated synthesis of type I collagen comprised of alpha1 and alpha2 chains at the injection site. These preliminary data demonstrate that collagen genes can be transferred into bone marrow stromal cells as well as fibroblasts in vivo and that the genes are efficiently expressed. These data encourage further studies in gene replacement for some forms of OI and use of bone marrow stromal cells as vehicles to deliver therapeutic genes to bone. PMID- 11500957 TI - ERK 1,2 and p38 pathways are involved in the proliferative stimuli mediated by urokinase in osteoblastic SaOS-2 cell line. AB - Bone metastases from prostate origin generate an osteoblastic reaction that is expressed in vitro by increased osteoblast proliferation. The urokinase-like plasminogen activator (u-PA) present in the media conditioned by tumoral prostatic cells acting as a ligand of the cellular membrane receptor (u-PAR), has been identified as the specific factor that modulates this proliferative reaction. The present study represents an effort to unravel the intracellular pathway by which u-PA activates osteoblastic proliferation and to evaluate the role of cellular receptor u-PAR in this proliferative phenomenon. Our results show that in vitro u-PA stimulates proliferation of SaOS-2 osteoblastic cells by activating the MAP kinase route of ERK 1 and 2 and the p38 pathway. These results are in accordance with the inhibition of intermediate activation and cell proliferation by PD 098059 and SB 203580, specific inhibitors of MEK and p38, respectively. We also show that SaOS-2 cells increase their proliferative response when cells are plated onto vitronectin, the second natural ligand of u PAR, and that culturing SaOS-2 cells in the presence of u-PA represents a stimuli for u-PAR expression. On the basis of these results we propose that osteoblastic cells respond to the prostate-derived u-PA stimuli in a very efficient manner that includes the utilization of two different signaling routes and the stimulation of the expression of the u-PA receptor. PMID- 11500958 TI - Characterization of the biologically important interaction between troponin C and the N-terminal region of troponin I. AB - The N-terminal regulatory region of Troponin I, residues 1-40 (TnI 1-40, regulatory peptide) has been shown to have a biologically important function in the interactions of troponin I and troponin C. Truncated analogs corresponding to shorter versions of the N-terminal region (1-30, 1-28, 1-26) were synthesized by solid-phase methodology. Our results indicate that residues 1-30 of TnI comprises the minimum sequence to retain full biological activity as measured in the acto S1-TM ATPase assay. Binding of the TnI N-terminal regulatory peptides (TnI 1-30 and the N-terminal regulatory peptide (residues 1-40) labeled with the photoprobe benzoylbenzoyl group, BBRp) were studied by gel electrophoresis and photochemical cross-linking experiments under various conditions. Fluorescence titrations of TnI 1-30 were carried out with TnC mutants that carry a single tryptophan fluorescence probe in either the N- or C-domain (F105W, F105W/C domain (88-162), F29W and F29W/N domain (1-90)) (Fig. 1). Low Kd values (Kd < 10(-7) M) were obtained for the interaction of F105W and F105W/C domain (88-162) with TnI 1-30. However, there was no observable change in fluorescence when the fluorescence probe was located at the N-domain of the TnC mutant (F29W and F29W/N domain (1 90)). These results show that the regulatory peptide binds strongly to the C terminal domain of TnC. PMID- 11500959 TI - Inhibitory effect of regucalcin on protein phosphatase activity in the nuclei of rat kidney cortex. AB - The role of regucalcin, which is a regulatory protein of calcium signaling, in the regulation of protein phosphatase activity in the nuclei of rat kidney cortex was investigated. Protein phosphatase activity towards phosphotyrosine, phosphoserine, and phosphothreonine was found in the nuclei. The enzyme activity towards three phosphoamino acids was significantly increased by the addition of calcium chloride (10-50 microM) in the enzyme reaction mixture. This increase was significantly inhibited by trifluoperazine (25 or 50 microM), an antagonist of calmodulin. The presence of regucalcin (50 or 100 nM) in the enzyme reaction mixture caused a significant decrease in protein phosphatase activity towards three phosphoamino acids. This effect was also seen in the presence of calcium (25 microM) and/or calmodulin (5 microg/ml). Protein phosphatase activity towards three phosphoamino acids was significantly increased in the presence of anti regucalcin monoclonal antibody (25 or 50 ng/ml) in the enzyme reaction mixture. This effect was completely blocked by the addition of regucalcin (100 nM). The effect of antibody (25 ng/ml) in increasing protein phosphatase activity towards phosphotyrosine was significantly inhibited by vanadate (10(-4) M). Also, the antibody's effect towards phosphoserine and phosphothreonine was significantly inhibited by cyclosporin A (10(-5) M). Endogenous regucalcin was found in the nuclei of rat kidney cortex using Western blot analysis. Nuclear regucalcin level was significantly reduced by the administration of saline (0.9% NaCl) for seven days in rats. Protein phosphatase activity towards three phosphoamino acids was significantly decreased by saline administration. The effect of anti-regucalcin monoclonal antibody (25 ng/ml) in increasing protein phosphatase activity towards three phosphoamino acids was weakened in the renal cortex nuclei of saline administrated rats. The present study demonstrates that endogenous regucalcin plays a suppressive role in the regulation of protein phosphatase activity in the nuclei of rat kidney cortex cells. PMID- 11500960 TI - Enhanced cartilage tissue engineering by sequential exposure of chondrocytes to FGF-2 during 2D expansion and BMP-2 during 3D cultivation. AB - Bovine calf articular chondrocytes, either primary or expanded in monolayers (2D) with or without 5 ng/ml fibroblast growth factor-2 (FGF-2), were cultured on three-dimensional (3D) biodegradable polyglycolic acid (PGA) scaffolds with or without 10 ng/ml bone morphogenetic protein-2 (BMP-2). Chondrocytes expanded without FGF-2 exhibited high intensity immunostaining for smooth muscle alpha actin (SMA) and collagen type I and induced shrinkage of the PGA scaffold, thus resembling contractile fibroblasts. Chondrocytes expanded in the presence of FGF 2 and cultured 6 weeks on PGA scaffolds yielded engineered cartilage with 3.7 fold higher cell number, 4.2-fold higher wet weight, and 2.8-fold higher wet weight glycosaminoglycan (GAG) fraction than chondrocytes expanded without FGF-2. Chondrocytes expanded with FGF-2 and cultured on PGA scaffolds in the presence of BMP-2 for 6 weeks yielded engineered cartilage with similar cellularity and size, 1.5-fold higher wet weight GAG fraction, and more homogenous GAG distribution than the corresponding engineered cartilage cultured without BMP-2. The presence of BMP-2 during 3D culture had no apparent effect on primary chondrocytes or those expanded without FGF-2. In summary, the presence of FGF-2 during 2D expansion reduced chondrocyte expression of fibroblastic molecules and induced responsiveness to BMP-2 during 3D cultivation on PGA scaffolds. PMID- 11500961 TI - Integrin alpha(v)beta(3) is involved in stimulated migration of vascular adventitial fibroblasts by basic fibroblast growth factor but not platelet derived growth factor. AB - We examined the effects of basic fibroblast growth factor (bFGF) and platelet derived growth factor (PDGF) on the migration of vascular adventitial fibroblasts (VAFs) isolated from rat aortic adventitiae. Both bFGF and PDGF significantly stimulated VAF migration in vitro. An antibody to rat beta(3) integrin reduced bFGF-stimulated migration in a dose dependent manner. Moreover, VAF migration was inhibited in the presence of cyclic RGD (cRGD) peptide. However, PDGF-directed migration was blocked only by equivalent cRGD peptide but not by antibody to beta(3) integrin. These data suggest that alpha(v)beta(3) integrin mediates VAF migration stimulated by bFGF and that chemoattractant directed migration may be through distinct integrins. PMID- 11500962 TI - G protein beta2 subunit antisense oligonucleotides inhibit cell proliferation and disorganize microtubule and mitotic spindle organization. AB - The association of G protein beta2 subunit (Gbeta2) with mitotic spindles in various mammalian cells has been demonstrated previously. Recently, we have identified the association of Gbeta2 protein with microtubules (Wu et al., [1998] J. Cell. Biochem. 70: 552-562). In the present experiment we have demonstrated the possible functional role of Gbeta2 in microtubule and mitotic spindle organization in mammalian cells. When Gbeta2 antisense phosphorothioate oligonucleotides were transfected into mammalian cells, inhibition of cell proliferation with cell death after a 4-day treatment was observed. If the transfected cells were incubated for two days and their Gbeta2 and microtubules were examined by Western blotting and immunofluorescence localization, marked reduction of the Gbeta2 protein, fragmentation and disassembly of cytoplasmic microtubules, and disorganized mitotic spindles were found. We conclude that the Gbeta2 protein is closely associated with microtubule assembly and may play a potential role in the regulation of cell proliferation and microtubule and mitotic spindle organization in mammalian cells. PMID- 11500963 TI - Stimulatory effect of lactate on testosterone production by rat Leydig cells. AB - Previously we found that the increased plasma testosterone levels in male rats during exercise partially resulted from a direct and luteinizing hormone (LH) independent stimulatory effect of lactate on the secretion of testosterone. In the present study, the acute and direct effects of lactate on testosterone production by rat Leydig cells were investigated. Leydig cells from rats were purified by Percoll density gradient centrifugation subsequent to enzymatic isolation of testicular interstitial cells. Purified rat Leydig cells (1 x 10(5) cells/ml) were in vitro incubated with human chorionic gonadotropin (hCG, 0.05 IU/ml), forskolin (an adenylyl cyclase activator, 10(-5) M), or 8-bromo-adenosine 3':5'-cyclic monophosphate (8-Br-cAMP, 10(-4) M), SQ22536 (an adenylyl cyclase inhibitor, 10(-6)-10(-5) M), steroidogenic precursors (25-hydroxy-cholesterol, pregnenolone, progesterone, and androstenedione, 10(-5) M each), nifedipine (a L type Ca(2+) channel blocker, 10(-5)-10(-4) M), or nimodipine (a potent L-type Ca(2+) channel antagonist, 10(-5)-10(-4) M) in the presence or absence of lactate at 34 degrees C for 1 h. The concentration of medium testosterone was measured by radioimmunoassay. Administration of lactate at 5-20 mM dose-dependently increased the basal testosterone production by 63-187% but did not alter forskolin- and 8 Br-cAMP-stimulated testosterone release in rat Leydig cells. Lactate at 10 mM enhanced the stimulation of testosterone production induced by 25-hydroxy cholesterol in rat Leydig cells but not other steroidogenic precursors. Lactate (10 mM) affected neither 30- nor 60-min expressions of cytochrome P450 side chain cleavage enzyme (P450scc) and steroidogenic acute regulatory (StAR) protein. The lactate-stimulated testosterone production was decreased by administration of nifedipine or nimodipine. These results suggested that the physiological level of lactate stimulated testosterone production in rat Leydig cells through a mechanism involving the increased activities of adenylyl cyclase, cytochrome P450scc, and L-type Ca(2+) channel. PMID- 11500964 TI - Assessment of promoter elements of the germ cell-specific proacrosin gene. AB - The testis-specific proacrosin gene encodes for a fertilization-promoting protein. In mouse and rat it is first transcribed in late pachytene spermatocytes and revealed to be translationally regulated. Former proacrosin promoter studies demonstrated that elements necessary for conducting a stage and temporal-specific expression of the gene are located within 0.9 kb upstream of the translational start codon. In the present study we analyzed putative cis-acting elements located in this promoter region for their specific binding properties to nuclear factors assumed to be involved in proacrosin gene regulation. Supplement of specific antibodies in electrophoretic mobility shift assays (EMSA) revealed that two Y-box proteins and the transcription factors CREM and YY1 interact with proacrosin promoter elements. The Y-box proteins, antigenically related to the frog Y-box proteins FRGY1 and FRGY2, bound to the Y-box (55-66 bp upstream of the ATG initiation codon) in brain and testis nuclear extracts, respectively. CREM bound to three elements (30-37, 252-259, and 717-724 bp upstream of ATG). The ubiquitous transcription factor YY1 bound to a conserved element in the central proacrosin promoter (457-473 bp upstream of ATG) and showed almost germ cell specific truncates in EMSA. These results suggest that the identified factors are involved in proacrosin gene regulation. PMID- 11500965 TI - Inhibition of connexin43 gap junctional intercellular communication by TPA requires ERK activation. AB - The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), is a potent inhibitor of gap junctional intercellular communication (GJIC). This inhibition requires activation of protein kinase C (PKC), but the events downstream of this kinase are not known. Since PKC can activate extracellular signal regulated kinases (ERKs) and these also downregulate GJIC, we hypothesized that the inhibition of GJIC by TPA involved ERKs. TPA treatment (10 ng/ml for 30 min) of WB-F344 rat liver epithelial cells strongly activated p42 and p44 ERK-1 and -2, blocked gap junction-mediated fluorescent dye-coupling, and induced connexin43 hyperphosphorylation and gap junction internalization. These effects were completely prevented by inhibitors of PKC (bis-indolylmaleimide I; 2 microM) and ERK activation (U-0126; 10 microM). These data suggest that ERKs are activated by PKC in response to TPA treatment and are downstream mediators of the gap junction effects of the phorbol ester. PMID- 11500966 TI - Statistical analysis of atomic contacts at RNA-protein interfaces. AB - Forty-five crystals of complexes between proteins and RNA molecules from the Protein Data Bank have been statistically surveyed for the number of contacts between RNA components (phosphate, ribose and the four bases) and amino acid side chains. Three groups of complexes were defined: the tRNA synthetases; the ribosomal complexes; and a third group containing a variety of complexes. The types of atomic contacts were a priori classified into ionic, neutral H-bond, C H...O H-bond, or van der Waals interaction. All the contacts were organized into a relational database which allows for statistical analysis. The main conclusions are the following: (i) in all three groups of complexes, the most preferred amino acids (Arg, Asn, Ser, Lys) and the less preferred ones (Ala, Ile, Leu, Val) are the same; Trp and Cys are rarely observed (respectively 15 and 5 amino acids in the ensemble of interfaces); (ii) of the total number of amino acids located at the interfaces 22% are hydrophobic, 40% charged (positive 32%, negative 8%), 30% polar and 8% are Gly; (iii) in ribosomal complexes, phosphate is preferred over ribose, which is preferred over the bases, but there is no significant preference in the other two groups; (iv) there is no significant prevalence of a base type at protein-RNA interfaces, but specifically Arg and Lys display a preference for phosphate over ribose and bases; Pro and Asn prefer bases over ribose and phosphate; Met, Phe and Tyr prefer ribose over phosphate and bases. Further, Ile, Pro, Ser prefer A over the others; Leu prefers C; Asp and Gly prefer G; and Asn prefers U. Considering the contact types, the following conclusions could be drawn: (i) 23% of the contacts are via potential H-bonds (including CH...O H bonds and ionic interactions), 72% belong to van der Waals interactions and 5% are considered as short contacts; (ii) of all potential H-bonds, 54% are standard, 33% are of the C-H...O type and 13% are ionic; (iii) the Watson-Crick sites of G, O6(G) and principally N2(G) and the hydroxyl group O2' is more often involved in H-bonds than expected; the protein main chain is involved in 32% and the side chains in 68% of the H-bonds; considering the neutral and ionic H-bonds, the following couples are more frequent than expected-base A-Ser, base G-Asp/Glu, base U-Asn. The RNA CH groups interact preferentially with oxygen atoms (62% on the main chain and 19% on the side chains); (iv) the bases are involved in 38% of all H-bonds and more than 26% of the H-bonds have the H donor group on the RNA; (v) the atom O2' is involved in 21% of all H-bonds, a number greater than expected; (vi) amino acids less frequently in direct contact with RNA components interact frequently via their main chain atoms through water molecules with RNA atoms; in contrast, those frequently observed in direct contact, except Ser, use instead their side chain atoms for water bridging interactions. PMID- 11500967 TI - Direct monitoring of molecular recognition processes using fluorescence enhancement at colloid-coated microplates. AB - Direct monitoring of recognition processes at the molecular level is a valuable tool for studying reaction kinetics to assess affinity constants (e.g. drugs to receptors) and for designing rapid single step immunoassays. Methods currently used to gain information about binding processes predominantly depend on surface plasmon resonance. These systems use excitation with coherent light in attenuated total reflection geometry to obtain discrimination between surface-bound and free molecules in solution. Therefore labeling of the compounds is not necessary, but due to the complexity of the measuring setup the method is rather costly. In this contribution we present a simple method for performing kinetic single step biorecognition assays with fluorophore labeled compounds using the fluorescence enhancement properties of surface bound silver colloids. Silver colloids are bound to standard microplates via silanization of the plastic surface. Fluorophores close to this colloid coated surface show a significant gain in fluorescence compared to fluorophores farther away in the bulk solution. Therefore discrimination between surface bound and free fluorophores is possible and the binding of, for example, fluorophore labeled antibodies to antigens immobilized on the colloid surface results in increasing fluorescence intensity. Utilization of standard microplates makes this method fully compatible with conventional microplate processing and reading devices. Neither excitation with coherent laser light nor ATR geometry is required, the measurement is performed in a standard fluorescence microplate reader in front face geometry with a xenon flash lamp as excitation source. Methods for the preparation of colloid-coated microplates and fluorescence-enhanced biorecognition assays are presented. Additionally the dependence of the system performance on the structure and properties of the metal colloid coated surface is described. A two-component biorecognition model system shows a detection limit in the subnanomolar range. The ease of colloid-surface preparation and the high sensitivity makes fluorescence enhancement at colloid-coated microplates a valuable tool for studying reaction kinetics and performing rapid single-step immunoassays. PMID- 11500968 TI - BIACORE J: a new platform for routine biomolecular interaction analysis. AB - SPR biosensor technology continues to evolve. The recently released platform from Biacore AB (Uppsala, Sweden), BIACORE J, is designed for the routine analysis of biomolecular interactions. Using an antibody-protein A and a ligand-receptor system, we demonstrate the utility of BIACORE J in determining active concentration and binding affinities. The results from these studies illustrate the high sensitivity of the instrument and its ability to generate reproducible binding responses. The BIACORE J is easy to operate and useful in diverse applications, making SPR technology widely accessible as a research tool. PMID- 11500969 TI - Binding of synthetic peptides by a human monoclonal IgM with an unusual combining site structure. AB - Using X-ray crystallography, a human monoclonal IgM cryoglobulin (Mez) was found to have an unusual combining site topography. Analysis of the unliganded Fv at 2.6 A resolution revealed that the HCDR3 had partitioned the active site into two compartments [Ramsland PA et al. 2000. Mol. Immunol. 37: 295-310]. The two cavities had dimensions and chemical properties that were compatible with the binding of peptides. In this study, libraries of peptides were prepared using solid-phase synthesis. Binding of the intact Mez IgM to these peptides was tested by enzyme-linked immunoassays. Screening of 400 dipeptides revealed that binding was markedly skewed toward amino acids with aromatic side-chains (Phe and Trp), especially when located in the second position. Preferential recognition of aromatic side-chains by Mez IgM was confirmed with larger peptides of three to five residues, but C-terminal positioning was not favored in these peptides. Mez IgM also showed binding propensities for acidic residues (Asp and Glu) as well as several other side-chains with different chemical properties, including His, Pro, Asn and Gln. Mez IgM recognized sets of overlapping octapeptides representing the sequences of the constant domains of human IgG1 heavy chains. These peptides represented similar stretches of polypeptide on the three-dimensional structures of all three constant domains (CH1, CH2 and CH3). Thus, Mez IgM may recognize structurally homologous regions of immunoglobulin domains, which were conserved during the evolution of the immune system. PMID- 11500970 TI - Localization of the large subunit of replication factor C near the 5' end of DNA primers. AB - Replication factor C (RFC) is a heteropentameric sliding clamp loader protein essential for processive synthesis of DNA by eukaryotic DNA polymerases delta and epsilon. To study the interaction of RFC with 3' and 5' ends of the DNA primer, we have developed chemical photocrosslinking assay using a synthetic DNA gap and DNA primer-template structures. We have found that the radioactively labeled primers containing a photoreactive group at their 5' end could crosslink with the largest RFC subunit (RFC140) on primer-templates and DNA gap structures, but that 3' end photoreactive primers could only crosslink with RFC140 within the DNA gap structure. Addition of replication protein A (RPA) to the reaction mixture resulted in the crosslinking of RPA subunits and inhibited crosslinking of RFC140 using 3' but not 5' photoreactive primers present at the gap. The results suggest specific contacts between RFC140 and the 5' end of the DNA primer. Together with previous data, these experiments allow us to propose a model for the DNA polymerase switch during eukaryotic DNA replication. PMID- 11500971 TI - IgM expressed by leukemic CD5(+) B cells binds mouse immunoglobulin light chain. AB - Mouse immunoglobulin (Ig) molecules have previously been shown to bind to the surface of CD5(+) B cells from patients with B-cell chronic lymphocytic leukemia (B-CLL). The results indicated that surface IgM was involved in the interaction and suggested the phenomenon was an example of the polyreactive binding capacity of the surface Ig (sIg) expressed by these malignant cells. This article describes the further characterization of the interaction between human IgM and mouse Ig molecules and subunits. Mouse Ig molecules of both kappa and lambda light chain classes bound to the B-CLL cell surface. The dissociation constant for the interaction of mouse IgG1 (K121) with the B-CLL cell surface was 3.6 x 10(-7) M. To confirm the involvement of the human IgM expressed by the B-CLL cells in the interaction, the malignant cells were stimulated in vitro to induce secretion of human IgM. Enzyme immunoassay was used to show that secreted human IgM bound to intact mouse Ig, as occurred with the cell surface analysis. The mouse Ig epitope recognized by the purified secreted human IgM was shown by Western blot analysis to be located on the light chain of the mouse Ig molecule and to be conformationally dependent. K121 light chain was cloned and expressed in E. coli and the recombinant light chain bound to the surface of CLL B cells. The results confirm that human IgM is the reactive ligand in the interaction with mouse Ig and indicate that the interaction of polyreactive IgM with mouse IgG occurs via the light chain component of IgG. PMID- 11500972 TI - Src homology-2 domain binding assays by scintillation proximity and surface plasmon resonance. AB - Various SH2 competitive binding assays, based on different techniques, have been described in the literature to identify and characterize SH2 ligands. The consideration that most reported methods show experimental limitations associated with assay parameters has prompted us to base our Src-SH2 inhibitor discovery program on the use of two different assays. In this study, two conceptually different biochemical methods designed to discover Src-SH2 inhibitors, respectively scintillation proximity assay (SPA) and surface plasmon resonance (SPR), have been evaluated and compared. For its high sensitivity and adaptability to automation SPA was chosen for high capacity screening (primary screen), whereas SPR was used for hits confirmation (secondary screening). However with the drastic improvement of inhibitor affinities, the limit of sensitivity was rapidly reached for the SPR assay based on the canonical pYEEI ligand. The substitution of the natural, monophosphorylated peptide ligand with a triphosphorylated peptide has allowed us to remarkably increase its sensitivity, so that molecules with nanomolar affinities could be easily differentiated in terms of IC(50) ranking. Such a new, improved SPR assay can be of great interest for the study of high affinity ligands of different SH2-based drug targets. PMID- 11500973 TI - Zebrafish mutants: behavioral genetic studies of visual system defects. AB - Zebrafish are a promising model for behavioral and genetic studies of vertebrate visual system development and retinal degeneration. In the past few years, numerous studies on zebrafish vision have been published. While most of the studies focus on the molecular and cellular characterization of mutations that disrupt zebrafish visual system structure in early development, others examine the mechanisms that underlie inherited visual system disorders in adults. Behavioral assays, along with morphologic and electrophysiological methods, are powerful tools for functional analyses of zebrafish visual development and performance. PMID- 11500974 TI - Agouti signaling protein and other factors modulating differentiation and proliferation of immortal melanoblasts. AB - The melanocyte lineage potentially forms an attractive model system for studies in cell differentiation, developmental genetics, cell signaling, and melanoma, because differentiated cells produce the visible pigment melanin. Immortal lines of murine melanoblasts (melanocyte precursors) have been described previously, but induction of differentiation involved a complex culture system with keratinocyte feeder cells. Here we describe conditions for both growth and induced differentiation of the melanoblast line melb-a, without feeder cells, and analyze factors that directly control proliferation and differentiation of these pure melanoblasts. Several active factors are products of developmental and other coat color genes, including stem cell factor (SCF), melanocyte-stimulating hormone (alphaMSH), and agouti signaling protein (ASP), a natural antagonist at the MSH receptor (melanocortin 1 receptor, MC1R) encoded by the agouti gene. A stable analog of alphaMSH (NDP-MSH) stimulated differentiation and inhibited growth. ASP in excess inhibited both effects of NDP-MSH, that is, ASP could inhibit pigmentation and stimulate growth. These effects provide an explanation for the interactions in mice of melanocyte developmental mutations with yellow agouti and Mc1r alleles, and a role for embryonic expression patterns of ASP. PMID- 11500975 TI - Cell proliferation and movement during early fin regeneration in zebrafish. AB - Cell proliferation and cell movement during early regeneration of zebrafish caudal fins were examined by injecting BrdU and Di-I, respectively. In normal fins of adult fish, a small number of proliferating cells are observed in the epidermis only. Shortly following amputation, epithelial cells covered the wound to form the epidermal cap but did not proliferate. However, by 24 hr, epithelial cells proximal to the level of amputation were strongly labeled with BrdU. Label incorporation was also detected in a few mesenchymal cells. Proliferating cells in the basal epithelial layer were first observed at 48 hr at the level of the newly formed lepidotrichia. At 72 hr, proliferating mesenchymal cells were found distal to the plane of amputation whereas more proximal labeled cells included mainly those located between the lepidotrichia and the basal membrane. When BrdU injected fins were allowed to regenerate for longer periods, labeled cells were observed in the apical epidermal cap, a location where cells are not thought to proliferate. This result is suggestive of cell migration. Epithelial cells, peripheral to the rays or in the tissue between adjacent rays, were labeled with Di-I and were shown to quickly migrate towards the site of amputation, the cells closer to the wound migrating faster. Amputation also triggered migration of cells of the connective tissue located between the hemirays. Although cell movement was induced up to seven segments proximal from the level of amputation, cells located within two segments from the wound provided the main contribution to the blastema. Thus, cell proliferation and migration contribute to the early regeneration of zebrafish fins. PMID- 11500976 TI - Endogenous electric current is associated with normal development of the vertebrate limb. AB - A steady ionic current is driven out of both developing and regenerating amphibian limbs. In the developing limbs of anurans and urodeles, focal outwardly directed current (0.5-2 microA/cm(2)) predicts the location of mesenchyme accumulations producing the early bud. Here, we report measurements of a similar outwardly directed ionic current associated with the development of the limb bud in the mouse and chick embryo by using a noninvasive, self-referencing electrode for the measurement of extracellular current. In both the mouse and chick embryo, flank currents were usually inwardly directed - the direction of Na(+) uptake by ectoderm. Outward currents associated with the mouse limb bud ranged from 0.04 10.8 microA/cm(2). Mouse limb bud and flank currents were similar to those measured in amphibian larvae, because they were reversibly collapsed and/or reversed by application of 30 microM amiloride, a Na(+) channel blocker. Unlike the amphibian embryos, flank ectoderm adjacent to the mouse limb bud in the anterior/posterior axis was usually associated with outwardly directed ionic current. This raises the possibility of a different, or changing, gradient of extracellular voltage experienced by mesenchyme cells in this plane of development than that observed in other regions of the limb bud. In the chick flank caudal to the somites, a striking reversal of the inwardly directed flank currents to very large ( approximately 100 microA/cm(2)) outwardly directed currents occurred three developmental stages before limb bud formation. We tested the relevance of this outwardly directed ionic current to limb formation in the chick embryo by reversing it by using an artificially applied "countercurrent" pulled through a microelectrode inserted just beneath the caudal ectoderm of the embryo. This application was performed for approximately 6 hr 2.5-3 developmental stages before hindlimb bud formation. This method resulted in abnormal limb formation by the tenth day of gestation in some embryos, whereas all control embryos developed normally. These data suggest an early physiological control of limb development. PMID- 11500977 TI - Differential response of Shh expression between chick forelimb and hindlimb buds by FGF-4. AB - The interactions of Sonic hedgehog (Shh) and fibroblast growth factor (FGF) play important roles in vertebrate limb pattern formation. In the posterior region of the chick limb bud, Shh and FGF-4 each maintain expression in a positive feedback loop. In the anterior region, Shh can also induce Fgf-4 expression in the anterior apical ectodermal ridge. However, the possibility of Shh induction by FGF protein is unclear. Because many experiments to analyze gene expression have been carried out by using the forelimb bud of the chick embryo, we investigated gene expression of the cells in the anterior region of the chick hindlimb bud after FGF-4 application and compared the results with those for the forelimb bud. When an FGF-4-containing bead was implanted into the anterior region of a stage 20 hindlimb bud, ectopic expression of Shh was induced in the mesenchyme beneath the anterior end of the apical ectodermal ridge at 36 hr after implantation. Subsequent to Shh activation, Hoxd13 was also observed in the anterior-distal region of the limb bud. Furthermore, FGF-4 implantation to the hindlimb bud caused additional digit formation accompanying respecification of positional value in the anterior tissue. Ectopic Shh was induced in cells located distal to the FGF-4 bead, and the cells of the flank region did not contribute to ectopic Shh induction. On the other hand, no ectopic Shh and Hoxd13 expression was detected by grafting an FGF-4 bead into the forelimb bud. Although FGF-4 implantation to the forelimb bud occasionally induced extra digit 2 formation, no embryos had an extra digit 3 or digit 4, and many specimens exhibited normal skeletal pattern. These results demonstrate the difference between the fore- and hindlimb buds in the cell competence of Shh induction in response to FGF-4, suggesting the possibility that the responsiveness of mesenchymal cells in signaling molecules is not the same in the fore- and hindlimb buds. PMID- 11500978 TI - Anterior expression of the caudal homologue cCdx-B activates a posterior genetic program in avian embryos. AB - Several families of regulatory genes have been implicated in anteroposterior patterning of gastrulation-stage vertebrate embryos. Members of the Drosophila caudal family of homeobox genes (Cdx) are among the earliest regulators of posterior cell fates. The regulatory cascade initiated by the caudal homologue, cCdx-B, was examined in avian embryos. During gastrulation, cCdx-B is expressed with other posterior patterning genes. In the posterior primitive streak, cCdx-B expression coincides with posteriorly expressed Hox cluster genes and Wnt family members such as Wnt-8c. The hierarchical relationship between these patterning genes was examined after anterior ectopic expression of cCdx-B. cCdx-B expression in anterior cardiogenic cells by means of adenoviral infection leads to the induction of Wnt-8c and the posterior Hox genes, Hoxa-7, Hoxc-6, and Hoxc-8. Cardiogenesis is not inhibited in cCdx-B expressing anterior lateral mesoderm, indicating that anterior cell fates are not respecified with the activation of posterior patterning genes after gastrulation. These results support an important role for cCdx-B in initiating a posterior program of gene expression that includes Wnt signaling molecules and the Hox cluster genes. PMID- 11500979 TI - Postembryonic expression of Caenorhabditis elegans mab-21 and its requirement in sensory ray differentiation. AB - The male tail sensory rays in Caenorhabditis elegans are complex copulatory structures, the normal patterning of which requires a number of regulatory genes. Among them, mab-21 specifies the identity of sensory ray 6. By using green fluorescent protein reporters, we identify multiple cis-acting elements that control the developmental expression of mab-21. Traced with a functional mab 21:gfp gene driven by authentic regulatory sequences, mab-21 expression could be detected in hypodermal, neuronal, muscle, and ray cells. We showed here that the expression of mab-21 in the hypodermis and neuronal cells was dispensable for its function in ray 6. In contrast, its expression in the ray 6 structural cell and neurons as conferred by its 3' enhancer was crucial for determining the correct ray 6 identity. PMID- 11500980 TI - Sorting nexin-14, a gene expressed in motoneurons trapped by an in vitro preselection method. AB - A gene-trap strategy was set up in embryonic stem (ES) cells with the aim of trapping genes expressed in restricted neuronal lineages. The vector used trap genes irrespective of their activity in undifferentiated totipotent ES cells. Clones were subjected individually to differentiation in a system in which ES cells differentiated into neurons. Two ES clones in which the trapped gene was expressed in ES-derived neurons were studied in detail. The corresponding cDNAs were cloned, sequenced, and analysed by in situ hybridisation on wild-type embryo sections. Both genes are expressed in the nervous system. One gene, YR-23, encodes a large intracellular protein of unknown function. The second clone, YR 14, represents a sorting nexin (SNX14) gene whose expression in vivo coincides with that of LIM-homeodomain Islet-1 in several tissues. Sorting nexins are proteins associated with the endoplasmic reticulum (ER) and may play a role in receptor trafficking. Gene trapping followed by screening based on in vitro preselection of differentiated ES recombinant clones, therefore, has the potential to identify integration events in subsets of genes before generation of mouse mutants. PMID- 11500981 TI - Neonatal estrogen exposure inhibits steroidogenesis in the developing rat ovary. AB - Treatment of newborn female rats with estrogens significantly inhibits the growth and differentiation of the ovary. To understand the molecular mechanism of estrogen action in the induction of abnormal ovary, we examined the expression profiles of steroidogenic factor 1 (SF-1) and several of its target genes in the developing ovaries after neonatal exposure to synthetic estrogen, estradiol benzoate (EB) by using reverse transcriptase polymerase chain reaction, in situ hybridization, and immunohistochemistry. Morphologic examination indicated inhibitory effects of estrogen on the stratification of follicles and development of theca and interstitial gland during postnatal ovarian differentiation. The expression of the steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage cytochrome P450 (P450(SCC)), which are both essential for steroid biosynthesis, markedly decreased in theca and interstitial cells throughout the postnatal development of the EB-treated ovary. However, expression of the transcriptional activator of the two genes, SF-1 was unaffected in theca and interstitial cells, although the number of these cells was lower in the EB treated ovary than in the control ovary. The expression of the estrogen mediator, estrogen receptor-alpha (ER-alpha), diminished specifically in theca cells at P6 and recovered by P14 in the EB-treated ovary. These results indicate that the effect of estrogens is mediated by means of ER-alpha resulting in the down regulation of StAR and P450(SCC) genes during early postnatal development of the ovary. These results suggest that the abnormal ovarian development by neonatal estrogen treatment is closely correlated with the reduced steroidogenic activity, and the data obtained by using this animal model may account in part the mechanism for aberrant development and function of the ovary in prenatally estrogen-exposed humans. PMID- 11500982 TI - TGFbeta Type III and TGFbeta Type II receptors have distinct activities during epithelial-mesenchymal cell transformation in the embryonic heart. AB - During the early stages of heart development, progenitors for the heart valves and septa come from endothelial cells via a developmental process known as "epithelial-mesenchymal cell transformation." This process is restricted to the atrioventricular (AV) canal and outflow tract portions of the embryonic heart. TGFbeta signal transduction pathways play critical roles during epithelial mesenchymal cell transformation in heart development. Previously, we showed that both TGFbeta Type II (TbetaRII) and Type III (TbetaRIII) receptors are required to mediate epithelial mesenchymal cell transformation in chick heart. Further, distinct TGFbeta2 and TGFbeta3 activities correspond to separate components of the embryonic cell transformation process. Studies by others of TGFbeta-mediated inhibition of cell proliferation produced a model where TbetaRIII functions by facilitating TGFbeta2 binding to TbetaRII. In the present study, we provide evidence that TbetaRIII mediates distinct cellular responses from those of TbetaRII. Blocking antibody for TbetaRIII, but not antibody against TbetaRII, specifically inhibits the endothelial cell-cell separation step. Examination of developmental markers, perturbed by blocking TbetaRIII antibody, revealed a pattern of expression distinctively different from that of TbetaRII antibody treatment. These data show that a distinct TbetaRIII-mediated process is required for endothelial cell-cell separation during epithelial mesenchymal cell transformation. As TGFbeta2 mediates endothelial cell-cell separation, the data point to a specific association of TGFbeta2 and TbetaRIII in the cell separation step of epithelial mesenchymal cell transformation. We conclude that distinct TbetaRII and TbetaRIII signal transduction pathways mediate epithelial mesenchymal cell transformation in the heart. PMID- 11500983 TI - Smoothelin expression during chicken embryogenesis: detection of an embryonic isoform. AB - Two isoforms of a novel smooth muscle cell (SMC) -specific cytoskeletal protein, smoothelin, have been described. In the adult chick, the 55-kDa smoothelin-A is expressed in visceral SMC, whereas the 120-kDa smoothelin-B is the major product in vascular SMC. Chicken was chosen to study smoothelin expression during embryogenesis and neonatally. Smoothelin-B was found in vascular SMC from stage 20 onward. In visceral SMC, smoothelin-B was present from stage 29 until hatching. Perinatally, a strong up-regulation of smoothelin synthesis was observed in visceral tissues, coinciding with a switch to the A-isoform. Transient smoothelin synthesis was observed in the somites and the developing heart. Western blotting revealed in these tissues a 62-kDa smoothelin isoform, designated smoothelin-C. Expression of the smoothelin isoforms seems to be strictly controlled with respect to cell type and developmental stage and may be related to the mode of contraction of the different cells. PMID- 11500984 TI - Protein and mRNA expression of serum and glucocorticoid-dependent kinase 1 in metanephrogenesis. AB - Abstract Expression of serum and glucocorticoid-dependent kinase 1 (SGK1) during development in mouse kidney (embryonic day [E] 14 to postnatal day [P] 1) was studied by in situ hybridization and immunofluorescence. In whole embryos, SGK1 mRNA was highly abundant in the developing metanephros, where SGK1 mRNA was expressed in the ureteric buds of the branching collecting duct system and in the mesenchymal blastema-derived comma- and s-shaped bodies. In E14 kidneys, SGK1 protein was below detection level, whereas at day E16, ureteric buds, s-shaped bodies and outgrowing loops of Henle expressed detectable amounts of SGK1 protein. SGK1 protein was also expressed in E16 primary tubules of the collecting duct system. In P1 kidneys, no or only faint SGK1 protein expression was apparent in comma- and s-shaped bodies, whereas SGK1 was continuously expressed by medullary collecting ducts. In conclusion, SGK1 is developmentally expressed in metanephrogenesis. High expression in developing collecting duct and in blastema derived comma- and s-shaped bodies suggests a dual function of SGK1 in maturation of the reabsorbing collecting duct epithelium and in epithelial transition of the blastema cells. PMID- 11500985 TI - Isolation and expression analysis of three zebrafish angiopoietin genes. AB - The Tie1 and Tie2 receptor tyrosine kinases and the Tie2 ligands, the angiopoietins, play critical roles in vertebrate vascular embryogenesis, helping to mediate the interaction between endothelial cells and the pericytes or vascular smooth muscle cells that envelop and support them. We have obtained full length cDNA sequences for zebrafish orthologs of angiopoietin-1 (ang1), angiopoietin-2 (ang2), and angiopoietin-like-3 (angptl3). Ang1 is expressed in head ventral mesenchyme, in the ventromedial region of somites, in mesenchyme surrounding trunk axial vessels, and in the hypochord, a transient embryonic structure of endodermal origin that has been implicated in dorsal aorta assembly in both zebrafish and Xenopus. Ang2 is expressed in head and anterior trunk ventral mesenchyme and the developing pronephric glomeruli. Angptl3 is expressed in the yolk syncytial layer. PMID- 11500986 TI - A system for the generation of curves on 3D brain images. AB - In this study, a computational optimal system for the generation of curves on triangulated surfaces representing 3D brains is described. The algorithm is based on optimally computing geodesics on the triangulated surfaces following Kimmel and Sethian ([1998]: Proc Natl Acad Sci 95:15). The system can be used to compute geodesic curves for accurate distance measurements as well as to detect sulci and gyri. These curves are defined based on local surface curvatures that are computed following a novel approach presented in this study. The corresponding software is available to the research community. PMID- 11500987 TI - Multiresolution analysis in fMRI: sensitivity and specificity in the detection of brain activation. AB - Multiresolution analysis of fMRI studies using wavelets is a new approach, previously reported to yield higher sensitivity in the detection of activation areas. No data are available, however, in the literature on the analytic approach and wavelet bases that produce optimum results. The present study was undertaken to assess the performance of different wavelet decomposition schemes by making use of a "gold standard," a realistic computer-simulated phantom. As activation areas are then known "a priori," accurate assessments of sensitivity, specificity, ROC curve area and spatial resolution can be obtained. This approach has allowed us to study the effect of different factors: the size of the activation area, activity level, signal-to-noise ratio (SNR), use of pre smoothing, wavelet base function and order and resolution level depth. Activations were detected by performing t-tests in the wavelet domain and constructing the final image from those coefficients that passed the significance test at a given P-value threshold. In contrast to previously reported data, our simulation study shows that lower wavelet orders and resolution depths should be used to obtain optimum results (in terms of ROC curve area). The Gabor decomposition offers the maximum fidelity in preserving activation area shapes. No major differences were found between other wavelet bases functions. Data pre smoothing increases ROC area for all but very small activation region sizes. PMID- 11500988 TI - Frequency dependence and gender effects in visual cortical regions involved in temporal frequency dependent pattern processing. AB - Neural response to flickering stimuli has been shown to be frequency dependent in the primary visual cortex. Controversial gender differences in blood oxygen level dependent (BOLD) amplitude upon 6 and 8 Hz visual stimulation have been reported. In order to analyze frequency and gender effects in early visual processing we employed a passive graded task paradigm with a dartboard stimulus combining eight temporal frequencies from 0 to 22 Hz in one run. Activation maps were calculated within Statistical Parametric Mapping, and BOLD amplitudes were estimated for each frequency within the striate and extrastriate visual cortex. The BOLD amplitude was found to steadily rise up to 8 Hz in BA 17 and 18 with an activation plateau at higher frequencies. In addition, we observed a laterality effect in the striate cortex with higher BOLD contrasts in the right hemisphere in men and in women. BOLD response rises similarly in men and women up to 8 Hz but with lower amplitudes in women at 4, 8, and 12 Hz (30% lower). No frequency effect above 1 Hz was found in the extrastriate visual cortex. There was also a regional specific gender difference. Men activated more in the right lingual gyrus (BA 18) and the right cerebellum compared with women, whereas women showed more activation in the right inferior temporal gyrus (BA 17). The study indicates that frequency dependent processing at the cortical level is limited to the striate cortex and may be associated with a more global information processing (right hemisphere dominance), particularly in men. The finding of significantly lower BOLD amplitudes in women despite previously shown larger VEP (visual evoked potential) amplitudes might suggest gender differences in cerebral hemodynamics (baseline rCBV, rCBF, or neurovascular coupling). The regional distinction points at additional differences in psychological processing even when using a simple visual stimulus. PMID- 11500989 TI - Differences in auditory processing of words and pseudowords: an fMRI study. AB - Although there has been great interest in the neuroanatomical basis of reading, little attention has been focused on auditory language processing. The purpose of this study was to examine the differential neuroanatomical response to the auditory processing of real words and pseudowords. Eight healthy right-handed participants performed two phoneme monitoring tasks (one with real word stimuli and one with pseudowords) during a functional magnetic resonance imaging (fMRI) scan with a 4.1 T system. Both tasks activated the inferior frontal gyrus (IFG), the posterior superior temporal gyrus (pSTG) and the inferior parietal lobe (IPL). Pseudoword processing elicited significantly more activation within the posterior cortical regions compared with real word processing. Previous reading studies have suggested that this increase is due to an increased demand on the lexical access system. The left inferior frontal gyrus, on the other hand, did not reveal a significant difference in the amount of activation as a function of stimulus type. The lack of a differential response in IFG for auditory processing supports its hypothesized involvement in grapheme to phoneme conversion processes. These results are consistent with those from previous neuroimaging reading studies and emphasize the utility of examining both input modalities (e.g., visual or auditory) to compose a more complete picture of the language network. PMID- 11500990 TI - Fast realistic modeling in bioelectromagnetism using lead-field interpolation. AB - The practical use of realistic models in bioelectromagnetism is limited by the time-consuming amount of numerical calculations. We propose a method leading to much higher speed than currently available, and compatible with any kind of numerical methods (boundary elements (BEM), finite elements, finite differences). Illustrated with the BEM for EEG and MEG, it applies to ECG and MCG as well. The principle is two-fold. First, a Lead-Field matrix is calculated (once for all) for a grid of dipoles covering the brain volume. Second, any forward solution is interpolated from the pre-calculated Lead-Fields corresponding to grid dipoles near the source. Extrapolation is used for shallow sources falling outside the grid. Three interpolation techniques were tested: trilinear, second-order Bezier (Bernstein polynomials), and 3D spline. The trilinear interpolation yielded the highest speed gain, with factors better than x10,000 for a 9,000-triangle BEM model. More accurate results could be obtained with the Bezier interpolation (speed gain approximately 1,000), which, combined with a 8-mm step grid, lead to intrinsic localization and orientation errors of only 0.2 mm and 0.2 degrees. Further improvements in MEG could be obtained by interpolating only the contribution of secondary currents. Cropping grids by removing shallow points lead to a much better estimation of the dipole orientation in EEG than when solving the forward problem classically, providing an efficient alternative to locally refined models. This method would show special usefulness when combining realistic models with stochastic inverse procedures (simulated annealing, genetic algorithms) requiring many forward calculations. PMID- 11500991 TI - Event-related changes in neuromagnetic activity associated with syncopation and synchronization timing tasks. AB - For low rhythmic rates (1.0 to approximately 2.0 Hz), subjects are able to successfully coordinate finger flexion with an external metronome in either a syncopated (between the beats) or synchronized (on each beat) fashion. Beyond this rate, however, syncopation becomes unstable and subjects spontaneously switch to synchronization to maintain a 1:1 stimulus/response relationship. We used a whole-head magnetometer to investigate the spatiotemporal dynamics of neuromagnetic activity (MEG) associated with both coordinative patterns at eight different rates spanning the range 1.0-2.75 Hz. Timing changes in the event related fields accompanied transitions from syncopation to synchronization and followed the placement of the motor response within each stimulus/response cycle. Decomposition of event-related fields into component auditory and motor brain responses revealed that the amplitude of the former decreased with increasing coordination rate whereas the motor contribution remained approximately constant across all rates. Such an interaction may contribute to changes in auditory-motor integration that cause syncopation to become unstable. Examination of event related changes in high frequency bands revealed that MEG signal power in the beta band (15-30 Hz) was significantly lower during syncopated coordination in sensors covering the contralateral sensorimotor area suggesting a dependence of beta rhythm amplitude on task difficulty. Suppression of beta rhythms was also stronger during synchronization preceded by syncopation, e.g., after subjects had switched, when compared with a control condition in which subjects synchronized throughout the entire range of rates. PMID- 11500992 TI - Non-stationary distributed source approximation: an alternative to improve localization procedures. AB - Localization of the generators of the scalp measured electrical activity is particularly difficult when a large number of brain regions are simultaneously active. In this study, we describe an approach to automatically isolate scalp potential maps, which are simple enough to expect reasonable results after applying a distributed source localization procedure. The isolation technique is based on the time-frequency decomposition of the scalp-measured data by means of a time-frequency representation. The basic rationale behind the approach is that neural generators synchronize during short time periods over given frequency bands for the codification of information and its transmission. Consequently potential patterns specific for certain time-frequency pairs should be simpler than those appearing at single times but for all frequencies. The method generalizes the FFT approximation to the case of distributed source models with non-stationary time behavior. In summary, the non-stationary distributed source approximation aims to facilitate the localization of distributed source patterns acting at specific time and frequencies for non-stationary data such as epileptic seizures and single trial event related potentials. The merits of this approach are illustrated here in the analysis of synthetic data as well as in the localization of the epileptogenic area at seizure onset in patients. It is shown that time and frequency at seizure onset can be precisely detected in the time frequency domain and those localization results are stable over seizures. The results suggest that the method could also be applied to localize generators in single trial evoked responses or spontaneous activity. PMID- 11500993 TI - Functional neuroanatomy of visuo-spatial working memory in Turner syndrome. AB - Turner syndrome (TS), a genetic disorder characterized by the absence of an X chromosome in females, has been associated with cognitive and visuo-spatial processing impairments. We utilized functional MRI (fMRI) to investigate the neural substrates that underlie observed deficits in executive functioning and visuo-spatial processing. Eleven females with TS and 14 typically developing females (ages 7-20) underwent fMRI scanning while performing 1-back and 2-back versions of a standard visuo-spatial working memory (WM) task. On both tasks, TS subjects performed worse than control subjects. Compared with controls, TS subjects showed increased activation in the left and right supramarginal gyrus (SMG) during the 1-back task and decreased activation in these regions during the 2-back task. In addition, decreased activation in the left and right dorsolateral prefrontal cortex (DLPFC) and caudate nucleus was observed during the 2-back task in TS subjects. Activation differences localized to the SMG, in the inferior parietal lobe, may reflect deficits in visuo-spatial encoding and WM storage mechanisms in TS. In addition, deficits in the DLPFC and caudate may be related to deficits in executive function during WM performance. Together these findings point to deficits in frontal-striatal and frontal-parietal circuits subserving multiple WM functions in TS. PMID- 11500994 TI - Influence of EEG electrodes on the BOLD fMRI signal. AB - Measurement of the EEG during fMRI scanning can give rise to image distortions due to magnetic susceptibility, eddy currents or chemical shift artifacts caused by certain types of EEG electrodes, cream, leads, or amplifiers. Two different creams were tested using MRS and T2* measurements, and we found that the one with higher water content was superior. This study introduces an index that quantifies the influence of EEG equipment on the BOLD fMRI signal. This index can also be used more generally to measure the changes in the fMRI signal due to the presence of any type of device inside (or outside) of the field of view (e.g., with fMRI and diffuse optical tomography, infrared imaging, transcranial magnetic stimulation, ultrasound imaging, etc.). Quantitative noise measurements are hampered by the normal variability of functional activation within the same subject and by the different slice profiles obtained when inserting a subject multiple times inside a MR imaging system. Our measurements account for these problems by using a matched filtering of cortical surface maps of functional activations. The results demonstrate that the BOLD signal is not influenced by the presence of EEG electrodes when using a properly constructed MRI compatible recording cap. PMID- 11500995 TI - Paradigm-dependent modulation of event-related fMRI activity evoked by the oddball task. AB - We have previously shown that event-related functional magnetic resonance imaging (ER-fMRI) may be used to record responses to the rapid, interleaved presentation of stimuli in the three-stimulus oddball task. The present study examined the sensitivity of ER-fMRI responses to variations in the range of inter-stimulus intervals (ISIs, calculated as the time from the offset of one stimulus to the onset of the next stimulus) and the type of behavioral response task used. ISIs were varied between a wide ISI range (550-2,050 msec) and a narrow ISI range (800 1,200 msec), while maintaining a similar mean ISI (approximately 1 stimulus per sec) between experiments. The response task was varied between button press and subvocal target counting. Gradient echo, echo planar images were acquired for each of three experiments (wide ISI with button press, narrow ISI with button press, and wide-ISI with counting) in five subjects. Target stimuli generated increased fMRI signal in a wide range of brain regions. The use of a narrow ISI range generated a greater volume of subcortical activity and a reduced volume of cortical activity relative to a wide ISI range. The counting task generated a larger amplitude and longer lasting evoked response in brain regions that responded during all three experiments. Rare distractor stimuli evoked fMRI signal change primarily in orbitofrontal, ventral-medial prefrontal and superior parietal cortex. These results illustrate that although ER-fMRI is relatively insensitive as a technique to small variations in the timing of stimulus-evoked responses, it is remarkably sensitive to consequences such variations have for the topographic location and amplitude of neural responses to stimuli. PMID- 11500996 TI - The Expert Consensus Guideline Series. Treatment of behavioral emergencies. AB - OBJECTIVES: Behavioral emergencies are a common and serious problem for consumers, their communities, and the healthcare settings on which they rely to contain, assess, and ultimately help the individual in a behavioral crisis. Partly because of the inherent dangers of this situation, there is little research to guide provider responses to this challenge. Key constructs such as agitation have not been adequately operationalized so that the criteria defining a behavioral emergency are vague. The significant progress that has been made for some disease states with better treatments and higher consumer acceptance has not penetrated this area of practice. A significant number of deaths of patients in restraint has focused government and regulators on these issues, but a consensus about key elements in the management of behavioral emergencies has not yet been articulated by the provider community. The authors assembled a panel of 50 experts to define the following elements: the threshold for emergency interventions, the scope of assessment for varying levels of urgency and cooperation, guiding principles in selecting interventions, and appropriate physical and medication strategies at different levels of diagnostic confidence and for a variety of etiologies and complicating conditions. METHOD: In order to identify issues in this area on which there is consensus, a written survey with 808 decision points was developed. The survey was mailed to a panel of 52 experts, 50 of whom completed it. A modified version of the RAND Corporation 9 point scale for rating appropriateness of medical decisions was used to score options. Consensus on each option was defined as a non-random distribution of scores by chi-square "goodness-of-fit" test. We assigned a categorical rank (first line/preferred choice, second line/alternate choice, third line/usually inappropriate) to each option based on the 95% confidence interval around the mean rating. Guideline tables were constructed describing the preferred strategies in key clinical situations. RESULTS: The expert panel reached consensus on 83% of the options. The relative appropriateness of emergency interventions was ascertained for a continuum of behaviors. When asked about the frequency with which emergency interventions (parenteral medication, restraints, seclusion) were required in their services, 47% of the experts reported that such interventions were necessary for 1%-5% of patients seen in their services and 32% for 6%-20%. In general, the consensus of this panel lends support to many elements of recent Health Care Financing Administration regulations, including the timing of clinician assessment and reassessment and the intensity of nursing care. However, the panel did not endorse the concept of "chemical restraint," instead favoring the idea that medications are treatments for target behaviors in behavioral emergencies even when the causes of these behaviors are not well understood. Control of aggressive behavior emerged as the highest priority during the emergency; however, preserving the physician-patient relationship was rated a close second and became the top priority in the long term. Oral medications, particularly concentrates, were clearly preferred if it is possible to use them. Benzodiazepines alone were top rated in 6 of 12 situations. High-potency conventional antipsychotics used alone never received higher ratings than benzodiazepines used alone. A combination of a benzodiazepine and an antipsychotic was preferred for patients with suspected schizophrenia, mania, or psychotic depression. There was equal support for high-potency conventional or atypical antipsychotics (particularly liquids) in oral combinations with benzodiazepines. Droperidol emerged in fourth place in some situations requiring an injection. CONCLUSIONS: To evaluate many of the treatment options in this survey, the experts had to extrapolate beyond controlled data in comparing modalities with each other or in combination. Within the limits of expert opinion and with the expectation that future research data will take precedence, these guidelines provide some direction for addressing common clinical dilemmas in the management of psychiatric emergencies and can be used to inform clinicians in acute care settings regarding the relative merits of various strategies. PMID- 11500997 TI - The Expert Consensus Guideline Series. Treatment of depression in women. AB - OBJECTIVES: Women constitute two-thirds of patients suffering from common depressive disorders. The treatment of depression in women is therefore a substantial public health concern. High-quality, empirical data on depressive disorders specific to women are limited. As a result, there are no comprehensive evidence-based practice guidelines on the best treatment approaches for these illnesses. We conducted a consensus survey of expert opinion on the treatment of 4 depressive conditions specific to women: premenstrual dysphoric disorder (PMDD), depression in pregnancy, postpartum depression in a mother choosing to breast-feed, and depression related to perimenopause/menopause. METHOD: After reviewing the literature and convening a work group of leading experts, we prepared a written survey covering a total of 858 treatment options in 117 specific clinical situations. Depression severity (mild to severe) was specified for most clinical situations. Treatment options included a broad range of pharmacological, psychosocial, and alternative medicine approaches. Most options were scored using a modified version of the RAND 9-point scale for rating appropriateness of medical decisions. We identified 40 national experts, 36 (90%) of whom completed the survey. Consensus on each option was defined as a non random distribution of scores by chi-square "goodness-of-fit" test. We assigned a categorical rank (first line/preferred choice, second line/alternate choice, third line/usually inappropriate) to each option based on the 95% confidence interval around the mean rating. Guideline tables indicating preferred treatment strategies were then developed for key clinical situations. RESULTS: The expert panel reached consensus on 76% of the options, with greater consensus in situations involving severe symptoms. For women with severe symptoms in each of the 4 central disorder areas we asked about, the first-line recommendation was for antidepressant medication combined with other modalities (generally psychotherapy), paralleling existing guidelines for severe depression in general populations. For milder symptoms in each situation, the panel was less uniform in recommending antidepressants. For the initial treatment of milder symptoms, the panel either gave equal endorsement to other treatment modalities (e.g., nutritional or psychobehavioral approaches in PMDD; hormone replacement in perimenopause) or preferred psychotherapy over medication (in conception, pregnancy, or lactation). In all milder cases, however, antidepressants were recommended as at least second-line options. Among antidepressants, selective serotonin reuptake inhibitors (SSRIs) as a class were recommended as first-line treatment in all situations. The specific SSRIs that were preferred depended on the particular clinical situation. Tricyclic antidepressants were highly rated alternatives to SSRIs in pregnancy and lactation. CONCLUSIONS: The experts reached a high level of consensus on the appropriateness of including both antidepressant medication, specifically SSRIs, and nonpharmacological modalities in treatment plans for severe depression in 4 key clinical situations unique to women. To evaluate many of the treatment options in this survey, the experts had to extrapolate beyond controlled data in comparing modalities with each other or in combination. Within the limits of expert opinion and with the expectation that future research data will take precedence, these guidelines provide some direction for addressing common clinical dilemmas in women. They can be used to inform clinicians and educate patients regarding the relative merits of a variety of interventions. PMID- 11500998 TI - Premenstrual dysphoric disorder: a guide for patients and families. PMID- 11500999 TI - Major depression during conception and pregnancy: a guide for patients and families. PMID- 11501000 TI - Postpartum depression: a guide for patients and families. PMID- 11501001 TI - Depression during the transition to menopause: a guide for patients and families. PMID- 11501002 TI - [Technical orthopedics: transforming synthetic aids to physical integration]. PMID- 11501003 TI - [Functional anatomy and kinematics of the femorotibial joint. Results of research 1836-1950]. AB - Ever since the investigations of the Weber brothers in 1836, on the mechanics of the walking implements of humans, several others, like von Meyer (1853), Langer (1858), Robert (1855), Albert (1878), Bougnion (1892), Fischer (1907), Zuppinger (1904), A.E. Fick (1877), R. Fick (1911), Strasser (1917), Tretter (1944), and Knese (1959), have presented a vast amount of literature on the kinematics and functional anatomy of the knee joint which continue to be a source of inspiration even for contemporary international research work. This literature has been reviewed and is now presented in short form, not only because of its historical value, but to stimulate further research activity in this field. Furthermore, possible application of the kinematics of the physiological knee joint in orthotics and prosthetics is indicated. PMID- 11501004 TI - [Syme amputation in heel defects]. AB - The main advantage of Syme's amputation is the end-bearing stump. A defective heel pad often leads to below-knee amputation. The question of interest is whether an atypical cutaneous flap also provides a covering for the stump adaptable to weight-bearing. Thirteen patients with heel pad ulcers or tumors who could not be attended to with a classic Syme's amputation were operated on in a modified fashion. Skin from the dorsum of the foot or a medial flap was used for covering the stump. Two patients underwent transtibial amputation. The remaining 11 patients received a covering for the end of the stump composed of tissue thick and bulky enough for weight-bearing in a prosthesis typical for a Syme stump. The results show that a transtibial amputation can be prevented by atypical soft tissue coverage of a Syme stump with satisfactory results regarding function and cosmetics. PMID- 11501005 TI - [Amputation of the lower extremity in paraplegic patients. Etiology, sequelae and patient management]. AB - Up to now, there are not many reports about indications and guidelines for prosthetic fitting in amputated paraplegics and tetraplegics. In a review of 35 spinal cord patients with 42 amputations of the lower extremity, data on the neurological level, cause of injury, level of amputation, long-term complications, prosthetic fitting, and usage of the prostheses in daily life are presented. Everyday usage of the prostheses is reported for 15 patients. PMID- 11501006 TI - [Coping with illness and psychotherapy for patients after amputation]. AB - Psychology is often neglected or glossed over in most works dealing with the rehabilitation of the amputee. Treating the psychological and social problems faced by the amputee often has more significance for his-satisfaction with life than the quality of the surgery or the nature of his prosthetic device. In the following paper, a complex model for the biopsychosocial adaptation after amputation is presented. In this model, the important diagnostic criteria in interaction with specific individual coping strategies are formulated, which explain the difficult psychosocial adaptation. Professional diagnosis and further indication are necessary on an individual basis. The main goal of every intervention is the psychic stabilization of the patient by taking into account his personal resources. Because of the complex somatic, social, and psychic problems which arise after an amputation, thorough cooperation between specialists is recommended. PMID- 11501007 TI - [Significance of static prosthesis alignment for standing and walking of patients with lower limb amputation]. AB - The influence of three alignment parameters of a transtibial prosthesis (sagittal foot position, plantar flexion, mediolateral foot position) on the load and motion of the lower extremity joints was investigated in 13 unilateral transtibial amputees. The aim was to determine whether a correlation exists between static prosthetic alignment and gait pattern that would allow an optimal biomechanical prosthetic alignment. The gait pattern was measured using kinematic, kinetic, and electromyographic methods. Statics was defined using the alignment apparatus L.A.S.A.R. Posture. The electromyogram of the m. vastus lateralis and m. biceps femoris was recorded on both sides. The motion of joints is described by joint angles. External joint moments define the mechanical loads. Alignment has almost no influence on muscle activity and joint mechanics of the contralateral leg. In contrast, prosthetic alignment affects clearly and systematically the load and motion of the knee joint during the stance phase on the ipsilateral side. The sagittal foot position influences the maximal flexion angle in the stance phase. The plantar flexion of the foot affects the temporal structure of knee motion. The mediolateral foot position causes correspondingly different varus and valgus moments acting on the knee. Swing phase motion does not depend on prosthetic alignment. The iEMG of the m. vastus lateralis is reduced. Innervation characteristics of the m. biceps femoris on the prosthetically fitted leg has completely changed. The ischiocrural muscles take over the neuromuscular action of the m. gastrocnemius to compensate for the external knee extension moment during the second part of the stance phase. Prosthetic statics determines if the knee joint is physiologically stressed in a standing posture and during walking. Statics will be correct if the anatomical knee axis of the standing amputee is located about 15 mm posterior to the load line in the sagittal plane. In the frontal plane, the load line touches the lateral patella border and strikes the middle of the foot about 5 cm anterior to the adapter. During walking, attention should be paid to performance of knee flexion in the stance phase. PMID- 11501008 TI - [Effect of walking speed on pressure distribution of orthopedic shoe technology]. AB - Lesions to the diabetic foot have various causes. However, there is broad consensus that excessive plantar pressure plays a major role in the chain of events leading to ulcerations and gangrenes. During walking, on the other hand, peak values of plantar pressure are likely to increase with velocity even in therapeutic shoes. Therefore, the question arises whether a moderate velocity should be recommended to diabetic patients to reduce the risk of foot lesions. In this study, two velocities were compared for different types of therapeutic footwear. The velocities selected were considered moderate (0.7 m/s) and normal (1.3 m/s) for diabetic patients. A specially designed mathematical algorithm (velocity normalization) provided the pressure distributions from a common set of measurements: seven trials at different velocities for each subject and each type of footwear. Ten test subjects with healthy feet were studied. The shoes were ready-made and all had a midfoot rocker. The following four conditions were tested: flexible or rigid outsole respectively in combination with a flat insole or molded foot bed respectively. Pressure distribution measurements were performed with the Pedar in-shoe system, and the Pedar software package was used for analysis. The foot was divided into six regions: first toe, second to fifth toes, metatarsal region, medial midfoot, lateral midfoot, and heel. Only peak pressures were taken into account. Gait velocity was found to have an effect on plantar pressure distribution, mainly in the toes and heel region. Peak pressure in the heels increased significantly by about 20%. In the toe region, the increase was about the same, but was not statistically significant. At a higher velocity, pressure even slightly decreased in the midfoot region. The percentage variation was similar for all four conditions. Thus, walking slowly prevented the foot from high peak pressures, and the combination of rigid outsole and molded foot bed was best suited for both slow and higher velocities. PMID- 11501009 TI - [Incidence and detection of nosocomial infections in technical orthopedics]. AB - The significance of nosocomial infections is increasing. The reasons for this are a higher concentration of problematic patients in hospitals due to the increasing number of outpatient surgeries, the increase of invasive therapeutic and diagnostic procedures, the growing share of immunodeficient patients as well as the increase in antibiotic-resistant and multiresistant pathogenic organisms. Basic changes in the medical system call for a systematic discussion about directed quality management. Surveillance, i.e., the systematic registration and evaluation of occurred diseases as well as the feedback to the personnel in charge is very important in this context, particularly when complemented by comprehensive hygienic measures. The growing significance of hospital infections increasingly triggered by multiresistant pathogenic organisms emphasizes the importance of general prevention. Each hospital has to determine which surveillance methods for the registration of nosocomial infections and multiresistant pathogenic organisms are the most appropriate under their respective conditions. To get a first idea of possible problem areas, prevalence studies are very suitable for the start before moving on to well-directed incidence studies in certain wards and for certain kinds of infection. PMID- 11501010 TI - [Biomechanical study of angle-stable anterior atlanto-axial spondylodesis plate]. AB - The optimum fixation method to achieve atlantoaxial fusion after odontoid resection is still subject to discussion. Isolated posterior surgical procedures for treatment of irreducible atlantoaxial kyphosis with spinal cord compression are associated with a high rate of morbidity and mortality. Transoral atlantoaxial plate fixation has been described by Harms as a fixation technique after odontoid resection. The purpose of this study was to compare biomechanically a new anterior atlantoaxial locking plate (AALP) with the Harms plate and the atlantoaxial transarticular screw fixation described by Magerl. Sixteen human cadaver craniocervical specimens were tested in flexion, extension, rotation, and bending with a nondestructive flexibility method using a nonconstrained testing apparatus. Five different groups were examined: (1) control group (intact) (n = 16), (2) unstable group (after dissection of the atlantoaxial ligaments and odontoidectomy) (n = 16), (3) Harms group (transoral atlantoaxial plate fixation) (n = 8), (4) AALP group (anterior atlantoaxial locking plate fixation) (n = 8), and (5) Magerl group (transarticular atlantoaxial screw fixation) (n = 16). Stiffness in any direction was significantly higher in the AALP specimens and those fixated with the Magerl method than in the Harms, control, or unstable specimens. The difference in stiffness between the AALP and Magerl reconstruction groups was not statistically significant. Experimentally, the AALP was significantly stiffer than the Harms plate. The AALP provided stiffness equal to transarticular screw fixation according to Magerl. A question that cannot be answered by this in vitro study concerns the level of rigidity required to obtain long-term stability and fusion by atlantoaxial fixation methods. However, it may be assumed that the more spinal motion is eliminated the greater the chance of definite spinal fusion. Therefore, it seems reasonable that the most reliable and rigid fixation method would be the method of choice. PMID- 11501011 TI - [Bone substitutes 2000 to 2010. Current status and innovation in therapy of bone defects]. PMID- 11501012 TI - [Artificial hand with innovative motoricity]. PMID- 11501013 TI - Analysis of relationship between Anopheles subpictus larval densities and environmental parameters using Remote Sensing (RS), a Global Positioning System (GPS) and a Geographic Information System (GIS). AB - Remote Sensing (RS), a Global Positioning System (GPS) and a Geographic Information System (GIS) were used to analyze relationship between Anopheles subpictus larval densities and environmental parameters in the Sekotong district on Lombok Island, Indonesia. Distance from the coast to larval habitats, season and surface water were considered as environmental parameters for determining An. subpictus larval densities. Japanese Earth Resources Satellite (JERS) Visible and Near Infrared Radiometer (VNIR) satellite imagery for the area acquired by National Space Development Agency of Japan (NASDA) were used to detect water, which could be used to characterize larval habitats. Data on larval sampling sites obtained from a GPS were entered into a GIS for mapping larval habitats to measure distance between the coast and the larval habitats. A GIS was used for overlaying of data coverages (i.e., water distribution from RS data and larval habitats coupled with data on larval densities) to identify factors that may explain the spatial distribution patterns of larval densities. An. subpictus larval densities were significantly associated with season and distance from the coast to larval habitats. The rainy season and the distance from the coast to larval habitats were critical environmental determinants for presence of An. subpictus larvae in the study. In this paper, we investigated relationship between An. subpictus larval densities and the environmental parameters using RS/GPS/GIS to determine if these tools could be used to predict larval densities. PMID- 11501014 TI - Immunolocalization of relaxin in the corpus luteum of the dromedary camel. (Camelus dromedarius). AB - The objective of this study was to determine the distribution of the peptide hormone relaxin in ovarian tissues of the pregnant and non-pregnant dromedary (Camelus dromedarius). Using light and electron microscopy we here describe the presence of relaxin in cells of the corpus luteum at various stages of the reproductive cycle of this important species. PMID- 11501015 TI - The effect of MS-818, newly synthesized pyrimidine compound, on fracture repair. AB - Both periosteal cell proliferation and endochondral ossification accompanied are characteristic of the fracture healing process. This process is regulated by various kinds of soluble factors including prostanoids, cytokines and growth factors. In particularly, basic fibroblast growth factor (bFGF) stimulates the mesenchymal cell proliferation and differentiation in the periosteum and leads to the fracture healing. Recently, newly synthesized pyrimidine compound, 2 piperadino-6-methyl-5-oxo-5,6-dihydro (7H) pyrrolo [3,4-d] pyrimidine maleate (MS 818) has been reported to augment the biological effect of bFGF in vitro. Therefore, we have studied the effect of MS-818 on fracture healing process in which bFGF has been reported to play an important role. In the rat fracture model, 5 mg/kg MS-818 which had been administered intraperitoneally for fourteen consecutive days enhanced the cartilage matrix formation. In the bone defect model, in which we can find only membranous ossification without chondrogenesis, cartilage matrix formation was observed in seven days after 1 microgram of human bFGF containing polymer pellet was embedded in the defect site. Chondrogenesis induced by bFGF was enhanced significantly after 5 micrograms MS-818 containing pellet was implanted with bFGF pellet. These results suggest that MS-818 might promote the fracture healing process through enhancement of the effect of bFGF on endochondral ossification. PMID- 11501016 TI - Intervertebral disc cell apoptosis by nitric oxide: biological understanding of intervertebral disc degeneration. AB - While the unphysiological mechanical load is a central etiologic factor of disc degeneration, biologic factors including nitric oxide (NO) seems to play an important role in this condition. It is, therefore, investigated whether NO is related to the degeneration of intervertebral disc by way of inducing the disc cell apoptosis. Twelve herniated lumbar disc and eight control specimens were obtained from the patients underwent surgery. Apoptotic cells were identified by TUNEL procedure and the percent apoptotic cell index (ACI%) of each sample was calculated. Detection of iNOS expression was performed by immunohistochemical analysis. Disc cell monolayer culture was prepared from the surgical specimen of the patients with lumbar disc herniation. NO generation of the disc cells was measured by Griess reaction. Cell proliferation was detected by measuring the incorporation of 3H-Thymidine. The extent of fragmented DNA induced by NO donor, NOC-18, was also measured by an enzyme-linked immunosorbent assay. The incidence of apoptotic cell death (ACI%) was greater in the herniated group (61.3 +/- 24.5%) than that of control group (5.6 +/- 6.8%; P < 0.001). iNOS positive disc cells were detected in all the samples. NO production of disc cells was enhanced by the stimulation of IL-1 alpha. Suppression of 3H-Thymidine incorporation and DNA fragmentation in the disc cells were promoted by treatment of 100 microM NOC 18. These results suggest that disc cells are able to release NO and NO may play an important role in the pathogenesis of disc degeneration through the induction of apoptosis of disc cells in situ. PMID- 11501017 TI - [Functional reconstruction of the hand after electrical injury with a composite flap of the vessel nerve and fascia]. AB - OBJECTIVE: In the case of electrical injury of the wrist, it may be necessary to transplant a long tendon, nerve and vessel for functional reconstruction. But the fibrosis of the recipient bed often makes the survival and functioning of the grafts difficult. This clinical study was to solve the problem. METHODS: A composite flap was designed consisting of the small saphenous vein, the sural nerve and the fascia of the lower-leg. The small saphenous vein and the nerve were transplanted to repair the defects of the radial or ulnar artery and the media nerve. The fascia was used to reconstruct the recipient bed for tendon grafting. RESULTS: Six cases were treated with this method with success in 5 cases. 5 to 18 months' follow-up revealed restoration of sensation and finger flexion in the 5 cases. CONCLUSION: This method is effective in alleviating tendon adhesion, accelerating functional restoration of the injured hand. PMID- 11501018 TI - [Anatomical study and clinical applications of the tensor fascia lata myocutaneous flap]. AB - OBJECTIVE: To explore the causes of blood circulation problems in 3 of 6 cases of pedicled tensor fascia lata myocutaneous flap transferring. METHODS: Anatomical study was performed on 16 cadavers (32 sides) after red emulsion injection. RESULTS: On 22 sides of 12 cadavers, the nutrient arteries lie in the muscle; on 2 sides of 2 cadavers, intramuscular nutrient arteries coexist with the big branches anterior to the muscle; on 4 sides of 3 cadavers, intramuscular nutrient arteries coexist with the big branches in the loose connective tissue deep to the muscle; on 4 sides of 3 cadavers, the nutrient arteries locate in the subcutaneous layer. According to the anatomical observations, much attention were paid to protect the vessels deep, superficial and anterior to the muscle during surgery, which resulted in successful clinical applications in 8 cases. CONCLUSIONS: The supplying arteries of the tensor fascia lata myocutaneous flap may exit not only intramuscularly, but also deep, superficial and anterior to the muscle. Special attention should be paid to them during operation. PMID- 11501019 TI - [Correction of blepharophimosis with a temporal periosteal flap]. AB - OBJECTIVE: To Provide a dependable and effective method for the treatment of blepharophimpsos. METHODS: In Blaskovics canthoplasty, a periosteum flap raised from the lateral orbit margin was turned over and fixed to the end of superiou tarsur. The operation was performed on 11 patients (13 lids), with blepharophimosis. Of whom 8 patients (8 lids) were serious blepharophimosis associated with cryptophthalmus or hollow socket; 3 patients (5 lids) were moderate. The epicanthus in 3 cases (4 lids) was corrected by Spaeth's technique. RESULTS: The follow-up period was 10.5 months in average. 10 lids of blepharophimosis were cured; 2 were improved significantly and 1 improved. No severe complications were found. CONCLUSIONS: The periosteal flap acts not only as a palpebral supporting tissue but also forms lateral ligament. The corrective effects of this method for blepharophimosis are fairly good. PMID- 11501020 TI - [The clinical application of the reverse island skin flap supplied by the superficial sural artery]. AB - OBJECTIVE: To conform the reliability of the reverse island skin flap supplied by the superficial sural artery in clinical applications. METHODS: The distally based superficial sural artery flap was designed on the posterior aspect of the leg. 15 patients were treated with this flap, who had soft-tissue shortage of the lower third of the leg, the foot or the heel. The size of the flap ranged from 10 cm x 8 cm to 5 cm x 4 cm. RESULTS: All flaps survived. The results were satisfactory after 6 to 18 months' postoperative follow-up. CONCLUSIONS: The blood supply of this flap is reliable without sacrificing the major arteries. Flap elevation is easy and quick. This flap may provide a new method in reconstructioning the soft-tissue shortage of the lower leg, the foot and the heel. PMID- 11501021 TI - [Reconstruction of the mandible and soft tissue defects with the osteomyocutaneous free fibula flap]. AB - OBJECTIVE: In order to ascertain a material that is ideal both for reconstruction of the composite mandibular defects and for osseointegrated implantation. METHODS: Fifteen osteomyocutaneous fibula flaps were transferred for reconstruction of the mandible and surrounding soft tissues. The characters of the blood supply of the flap, the operation course, the relation of the fibula and dental implants were observed. RESULTS: The osteomyocutaneous fibula flap was supplied by double vessels. The blood supply was reliable even after the fibula was cut into several segments. Periosteal circulation was its main blood supply. The postoperative contour and function of the mandibular region were good. The flap also favours dental implants and denture. CONCLUSION: The osteomyocutaneous fibula flap was the ideal material to reconstruct the defects of mandible and surrounding soft tissues. PMID- 11501022 TI - [Expanded extra-long fascia flap in the trapezius region for repair of scar contracture of the neck]. AB - OBJECTIVE: In order to gain a larger fascia flap in the trapezius region and to close the donor site easily and primarily. METHODS: We applied tissue expansion to form the expanded extra-long trapezius fascia flap. RESULTS: The flap was used to repair the neck scar contracture with satisfactory effects. CONCLUSIONS: The flap is very useful for patients who have a large burned defect and are short of donor sites, especially the children. PMID- 11501023 TI - [Viability and dynamic observation of stretchback of the continuously expanded skin flap in the pig model]. AB - OBJECTIVE: To investigate the viability of the continuously expanded skin flap and the dynamic change of flap stretchback in the pig model. METHODS: White piglets and a constant pressure-infusing pump were employed to establish a continuous tissue expansion model. The animals were divided into two groups. The short rectangular expander of 180 ml was used in group A and the long rectangular expander of 210 ml was used in group B. Three days after completion of the continuous expansion, normal random and expanded skin flaps were created to assess flap viability and stretchback. RESULTS: Flap survival length in group A and B increased by 92.60% and 91.96% respectively versus normal random flap. But there was no significant difference in survival length between group A and B. Obvious flap stretchback was observed in the early stage. Eight weeks after flap operation the flap width of group A and B restored to that of the random skin flaps (P > 0.05). CONCLUSION: Continuously fast-expanded skin flaps are reliable in viability for soft tissue reconstruction. Larger expanders cannot further enhance the viability of the expanded flap. PMID- 11501024 TI - [Experimental study of biologic effects of tumour necrosis factor-alpha on scar derived fibroblasts]. AB - OBJECTIVE: To investigate the biologic effects of tumour necrosis factor alpha(TNF-alpha) on scar-derived fibroblasts and to evaluate the role of TNF alpha in genesis, prevention and treatment of scar. METHODS: Cell strains were established by tissue culture technique. To study the fibroblasts derived from hypertrophic scar and normal dermis, cytoactivity and expression of fibronectin (FN) were determined by MTT, FCM and immunofluorescence. RESULTS: 1. When TNF alpha is at a high concentration (1,000 U/ml increases), the cytoactivity of hypertrophic scar fibroblasts was decreased significantly, and the positive rate of FN expression was decreased by 11.7%. 2. Cytoactivity of normal dermis fibroblasts was increased with TNF-alpha while the positive rate of FN expression was increased by 9.6%. CONCLUSIONS: TNF-alpha has different biological effects on fibroblasts derived from hypertrophic scar or normal dermis. Studying the regulative mechanisms of TNF on abnormal scar will probably provide a new way to prevent and cure it. PMID- 11501025 TI - [An experiment study on wound healing with exogenous hyaluronic acid]. AB - OBJECTIVE: To evaluate the effects and possible mechanisms of hyaluronic acid (HA) to inhibit scar formation as well as the dose-dependent relationship. METHODS: Forty-five health adult SD rats in a dorsal trauma model were randomly divided into the 2% HA treated-group(group A), 1% HA treated-group(group B), and the PBS treated-group as a control group(group C). The process of healing was observed, the remained wound areas were measured, and a mean wound healing time was calculated. Furthermore, the dynamic analysis was performed on local tissue protein, HPr and PCIII contents. RESULTS: 1. The remained wound areas of the control group were much smaller than that of group A and B, and the mean value of wound healing time was 15.2 +/- 2.0, 14.2 +/- 1.6 and 13.4 +/- 1.4 days respectively. 2. Histologically, when compared qualitatively with PBS controls, HA-treated wounds exhibited a less inflammatory response and angiogenesis. The fibroblast proliferated significantly, and the collagen fibril was slender and arrayed in good order, 3. The contents of tissue HPr were statistically lower in HA-treated groups, while the contents of PCIII were statistically higher compared to the control group. CONCLUSIONS: The study indicated that 1. The exogenous hyaluronan acid may effectively inhibit collagen synthesis and increase tissue PCIII content. As a result, the scar formation was decreased. 2. Hyaluronal efficacy presented a dose-dependent manner. PMID- 11501026 TI - [Quantitative study on the expression of mRNA for TGF-beta and collagenase (MMP 1), tissue metalloproteinase inhibitor-1 (TIMP-1) in hypertrophic scar]. AB - OBJECTIVE: To investigate the expression of mRNA for TGF-beta, MMP-1 and TIMP-1 and their effects on the formation of hypertrophic scar (HTS). METHODS: In situ hybridization and image analysis were used in this study. RESULTS: The expressions of mRNA for TGF-beta and TIMP-1 in HTS was greatly increased compared with those of normal skin. The average integral optical density of mRNA for TGF beta and TIMP-1 increased by 8 and 7 folds (P < 0.001) related to normal skin and the expression of them showed highly positive correlation. CONCLUSIONS: The findings suggest that TGF-beta promotes the expression of TIMP-1 and may be a possible factor for hypertrophic scar formation. PMID- 11501027 TI - [Expression of alpha 1-4 integrins in hypertrophic scar fibroblasts]. AB - OBJECTIVE: To determine the expression levels of alpha 1-4 integrins in hypertrophic scar fibroblasts and investigate the role that alpha 1-4 integrins may play in the process of hypertrophic scar formation. METHODS: The membrane protein was extracted from hypertrophic scar and normal skin tissues. The content of alpha 1-4 integrins of the membrane protein was detected by immunoblotting method. The locations of alpha 1-4 integrin expression were examined by immunohistochemical stain. RESULTS: 1. Higher concentration of alpha 1-4 integrins was identified in the membrane protein extracted from hypertrophic scar than from normal skin. 2. In normal skin, the expression of alpha 1, alpha 2 and alpha 3 integrins located on the epidermal cell surface but not on the dermal fibroblast surface. In the hypertrophic scar, not only did the epidermal cell surface express alpha 1-3 integrins, but also the fibroblast surface expressed alpha 1-4 integrins. CONCLUSIONS: The expressions of hypertrophic scar fibroblast surface integrins are subjected not only to cell-extracellular matrix adhesion, but also to intracellular/extracellular signal transduction. Integrins may play an important role in regulating much of fibroblast behavior, including growth, differentiation, phenotype, and in scar tissue remodelling. PMID- 11501028 TI - [The application of meshed acellular dermis xenograft (pigskin) with split thickness skin autograft in 23 patients]. AB - OBJECTIVE: An effective and inexpensive coverage, which can prevent scar formation and fill up scar tissue depression for full thickness skin burns in 23 patients was presented. METHODS: A deepithelized and acellular basement membranes of dermis from freshly harvested pigskin was obtained as meshed acellular dermis xenograft. The patients were treated in one or two stages. Two stages, 7 day postoperative the meshed acellular xenograft, a split thickness skin autograft was applied. One stage, the meshed acellular xenograft and split thickness skin autograft were applied at same time. RESULTS: The take rate of two staged treatment was (87.0 +/- 25.4)% and one stage was (60.3 +/- 3.4)%. All grafts showed in pink or near normal skin color and felt soft and smooth. CONCLUSIONS: The combination of meshed acellular dermis xenograft and split thickness skin autograft demonstrated an ideal way for the coverage of full thickness skin burn, which is effective and inexpensive. PMID- 11501029 TI - [A serial histological study on suture expansion osteogenesis for cleft palate closure]. AB - OBJECTIVE: The purpose of this study was to know the process of tissue response and regeneration in the palato-maxillary suture under tensile forces. METHODS: Sutural tissue changes after 3, 7, 14, 28, 56 days of suture expansion were examined histologically and fluorescently. RESULTS: It was found that a cyst-like zone appeared in the conjuncture of bony front and sutural connective tissue at the early stage of sutural expansion. Then a great amount of osteoblasts and fibroblasts proliferated. New bone deposited along the septum and the front of the cyst until the new bone front formed and the suture restored its original morphology. CONCLUSIONS: The findings suggested that sutural expansion be a process of bone tissue regeneration and fibrous tissue repair. PMID- 11501030 TI - [The effects of electric copper needle on venous thrombosis in the rat]. AB - OBJECTIVE: In order to study the relation between the electric copper needle and venous thrombosis in the rat. METHODS: 90 rats were divided into two groups, each using either the superficial epigastric vein or the femoral vein for the experiment. The relations of voltage, current, electrifying time and venous thrombosis were studied and the histological changes were observed. RESULTS: 1. 1.4 V DC did not damage the vessel-nerve. 2. The current declined with time to 0.1 mA. 3. When the diameters of the copper needle and the vein were similar (P > 0.05), the thrombogenesis time was from 5.5 to 10.5 minutes. When the diameters of the copper needle and the vein were different (P < 0.05), the thrombogenesis time was from 7.5 to 17.5 minutes. CONCLUSIONS: For the electric copper needle to get ideal treatment, the best voltage is 4 V DC for 5.5 to 10.5 minutes or 7.5 to 17.5 minutes. PMID- 11501031 TI - [Advances in the study of etiologic mechanism of keloid]. PMID- 11501032 TI - [Cosmetic surgery and science]. PMID- 11501033 TI - [Progress in the study of etiology and treatment of palpebral bags]. PMID- 11501034 TI - [Transposition of the pedicled latissimus dorsi myocutaneous flap for the treatment of lymphedema of the upper limb after radical mastectomy]. AB - OBJECTIVE: The clinical study was to improve the surgical treatment to upper limb lymphedema. METHODS: The author treated 12 cases of severe lymphedenia of the upper limb using the pedicled latissimus dorsi myocutaneous flap. The treatment was completed in one stage. RESULTS: The results were satisfactory in all the cases during the follow-up period of one to three years. No recurrence occurred. CONCLUSIONS: The method is a good treatment to secondary lymphedema of the upper limb after radical mastectomy as it possesses the advantages of casiness to perform, short operation time, reliable effects, very safety and accordance to physiology. PMID- 11501035 TI - Calcitonin gene-related peptide: an endogenous mediator of preconditioning. AB - Preconditioning of the heart induced by a brief ischemia or hyperthermia is exerted in two phases, early and delayed protection. The cardioprotection of ischemic preconditioning is related to the release of endogenous mediators. Calcitonin gene-related peptide (CGRP), a principal transmitter of capsaicin sensitive sensory nerves, is involved in the mediation of ischemic preconditioning, and CGRP-mediated ischemic preconditioning has been shown to protect the endothelial cells. The cardioprotection mediated by endogenous CGRP is also found in heat stress reaction. Drug-induced preconditioning, such as by nitroglycerin, may be related to stimulated release of CGRP. These findings suggest that CGRP may be an endogenous myocardial protective substance and plays an important role in the mediation of preconditioning. PMID- 11501036 TI - Prostaglandins but not nitric oxide are endothelium-derived relaxing factors in the trout aorta. AB - AIM: To identify the type of prostanoids produced by endothelial cells of trout aorta and to determine whether or not the smooth muscle responds to nitric oxide. METHODS: Ventral aortas, with and without endothelium from rainbow trout (S gairdneri), were incubated in a buffered salt solution. RESULTS: Addition of the calcium ionophore A23187 caused a significant increase in prostaglandin E's and a consistent increase in the stable metabolite of prostacyclin (6-keto prostaglandin F1 alpha) in the incubation media only when the endothelium was present. This production was inhibited by methylene blue (10 mumol/L). In rings of trout aorta without endothelium suspended for the measurement of isometric force in organ chambers, prostacyclin and prostaglandin E1 but not prostaglandin E2 caused concentration-dependent decreases in tension when the rings were contracted with acetylcholine. The smooth muscle did not relax to nitric oxide but did so to sodium nitroprusside. Relaxations to the latter nitrovasodilator were not inhibited by methylene blue. Descending aorta without endothelium from frogs relaxed in a concentration-dependent manner to nitric oxide. CONCLUSION: Predominant endothelium-derived relaxing factors in trout aorta are prostaglandins, the synthesis of which can be inhibited by methylene blue. A phylogenetic appearance of nitric-oxide sensitive mechanism for vasodilatation, perhaps is associated with the transition from water to air respiration. PMID- 11501037 TI - Effects of microiontophoretically-applied opioid peptides on Purkinje cells in the cat cerebellum. AB - AIM: The purpose of the present study was to examine the effects of microiontophoretically-applied opioid peptides on Purkinje cell of the cerebellum. METHODS: The effects of microiontophoretically-applied morphine, leucine-enkephalin (Leu-Enk), methionine-enkephalin (Met-Enk), and dynorphin 1-13 (Dyn) on the spontaneous discharge of Purkinje cells in the cerebellum of the anesthetized cat were examined. RESULTS: Microiontophoretic applications of Leu Enk and morphine produced inhibitory and excitatory responses, respectively in Purkinje cells. Application of both morphine and Leu-Enk induced dose-dependent responses. The excitatory responses were antagonized by naloxone, whereas the inhibitory responses were not. Bicuculline, a GABA-A antagonist, completely abolished both the Leu-Enk- and morphine-induced-inhibitory responses. Iontophoretic application of Met-Enk and dyn produced inhibitory responses only. Met-enk- and dyn-induced inhibition was antagonized by naloxone. CONCLUSION: In Purkinje cell activity, microiontophoretically applied Leu-Enk- and morphine induced excitation is connected with opiate receptors, whereas inhibition is related to the GABA receptor. However, Met-Enk and dyn produced only inhibitory effects via an opiate receptor in the cerebellum of cats. PMID- 11501038 TI - Relevance between striatal expression of Fos, proenkephalin mRNA, prodynorphin mRNA and rotation induced by l-stepholidine in 6-hydroxydopamine-lesioned rats. AB - AIM: To study that l-stepholidine (SPD) regulates the expression of proenkephalin (PENK) and prodynorphin (PDYN) mRNA and Fos in the striatum after rotational test in the 6-hydroxydopamine (6-OHDA)-lesioned rats. METHODS: PENK and PDYN mRNA levels were examined with in situ hybridization, and Fos expression was detected with immunocytochemistry. The data were semi-quantified with image analyzer. RESULTS: (1) Following repeated SPD treatment, the rotation was kept on high activity in the 6-OHDA-lesioned rats. (2) SPD significantly elicited Fos expression in both sides of striatum, particularly in the denervated one. Repeated administration of SPD, Fos expression declined on both sides, particularly in the intact one. (3) In the denervated striatum of 6-OHDA-lesioned rats, the PENK mRNA level was extremely increased vs that in the intact striatum. This high level of PENK mRNA was significantly reduced by 7-d treatments of SPD. SPD also reduced the level of PENK mRNA in the intact striatum. However, the level of PDYN mRNA did not show significant change in both sides of striatum after denervation or SPD treatment. CONCLUSION: In the 6-OHDA-lesioned rats, the rotation induced by SPD was kept on a high activity, which was in pace with the inducement of Fos expression and the reduction of expression of PENK mRNA in the denervated striatum. But then the lesion and SPD treatment had no remarkable effect on the expression of PDYN mRNA. PMID- 11501039 TI - Moclobemide-induced gynecomastia in rats. AB - AIM: To study the toxic effect of moclobemide on male breast and to elucidate its mechanism of action. METHODS: Routine histopathological analysis was used to diagnose the effect of moclobemide on male breast in rats. Plasma concentrations of estrogen, androgen, and prolactin were measured by a ratioimmunometer and relative receptors of mammary gland tissue were detected immunohistochemically. RESULTS: After 180-d moclobemide treatment, the presence of gynecomastia was 0, 5, 5, 7/10 rats in 0, 60, 240, and 600 mg/kg groups, respectively. After 30-d convalescence, only one rat in 600 mg/kg group got the incidence of gynecomastia. Serum prolactin concentration had a trend to decrease with increasing dose and prolactin receptors in mammary gland were up-regulated. CONCLUSION: Long-term treatment with moclobemide causes gynecomastia in rats, which is reversible. The mechanism of moclobemide-induced gynecomastia may be related to the increase in prolactin receptors in mammary glands. PMID- 11501041 TI - Effect of histamine H3-receptor antagonist clobenpropit on spatial memory of radial maze performance in rats. AB - AIM: The effect of histamine H3-receptor antagonist, clobenpropit (VUF9153) on spatial memory deficits induced by scopolamine was investigated in rats. METHODS: Eight-Arm radial maze performance was used to measure spatial memory in rats, and the brain regions were subsequently dissected and histamine contents were determined by HPLC. RESULTS: Intracerebroventricular (icv) injection of clobenpropit (50 micrograms) ameliorated memory impairment induced by scopolamine in both parameters of radial maze performance. The amelioration induced by clobenpropit was antagonized by an H3-agonist, (R)-alpha-methylhistamine (5-10 micrograms). alpha-Fluoromethylhistidine (10, 20 micrograms), a histidine decarboxylase inhibitor, also effectively reversed clobenpropit-induced ameliorating effects. CONCLUSION: The brain histamine H3-receptor antagonists are highly related to the spatial memory, and this action may be due to cholinergic neurons. PMID- 11501040 TI - Dithiolane analogs of lignans inhibit interferon-gamma and lipopolysaccharide induced nitric oxide production in macrophages. AB - AIM: To investigate the effect of a group of novel synthetic dithiolane analogs of lignans and a well characterized platelet-activating factor (PAF) receptor antagonist, L659,989 on PAF-receptor binding, IFN-gamma- and lipopolysaccharide (LPS)-induced NO production, and steady-state inducible nitric-oxide synthase (iNOS) mRNA expression. METHODS: PAF-receptor binding study was performed by displacement of 3H-PAF from rabbit platelet membrane; NO production was quantitated by measuring the NO oxidation product, nitrite, in conditioned culture medium; expression of iNOS mRNA was assessed by Northern blot analysis. RESULTS: The dithiolane analogs inhibited the production of NO, decreased iNOS mRNA expression and antagonized PAF-receptor binding. L659,989 had no effect on NO production and iNOS mRNA expression. Among the compounds tested, there was no simple correlation between their PAF-receptor antagonistic and iNOS inhibitory activities. CONCLUSION: The dithiolane analogs are a new synthetic chemical class of iNOS expression regulators with dual biologic functions: inhibiting iNOS induction and blocking PAF-receptor. PMID- 11501042 TI - Synaptic interaction between GABAergic terminals and substance P receptor positive neurons in rat spinal superficial laminae. AB - AIM: To identify synaptic relationship between gamma-aminobutyric acid (GABA) containing terminals and substance P receptor-positive neurons in the superficial dorsal horn of rat spinal cord. METHODS: A combination of preembedding immunoperoxidase staining for substance P receptor with postembedding immunogold staining for GABA was employed. RESULTS: Intense substance P receptor-expressing neurons were concentrated in medial half of the lamina I and the outer lamina II of the spinal cord. Under electron microscopy, substance P receptor immunoreactivity was found mainly in perikarya and dendrites, and deposited along neuronal membranes, in Golgi complex, in rough endoplasmic reticulum and on the surface of mitochondria with patch- or granular-like staining. The double labeling studies revealed that substance P receptor-expressing dendrites received symmetric synaptic contacts from axonal terminals labeled with immunogold particles indicating GABA. GABA also co-localized with substance P receptors in the dendrites. CONCLUSION: The synaptic contact between GABA-containing terminal and substance P receptor-expressing neuron provided important morphological evidence for previous pharmacological studies concerning antinociceptive function of GABA in the spinal cord. PMID- 11501043 TI - Effect of ginsenoside on IL-1 beta and IL-6 mRNA expression in hippocampal neurons in chronic inflammation model of aged rats. AB - AIM: To study the effects of ginseng root saponins (GRS) on interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) expression in hippocampal neurons in chronic inflammation model of aged rats. METHODS: Chronic inflammation was induced by consecutive intraperitioneal injections of bacterial lipopolysaccharides (LPS) in aged rats. The semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was used for determination of IL-1 beta and IL-6 mRNA in hippocampal neurons of aged rats. RESULTS: No marked expression of IL-1 beta and IL-6 mRNA was found in hippocampal neurons of young rats (3 months). In adult rats (9 months) the levels of IL-1 beta and IL-6 expressed were 1.54 +/- 0.21 and 1.63 +/ 0.21, respectively; in aged rats (24 months) treated with LPS the levels were 1.98 +/- 0.21 and 1.91 +/- 0.11 respectively, significantly higher than those of adult rats. After treatment with GRS (10, 20, 40 mg/kg), the IL-1 beta and IL-6 mRNA content in aged rats decreased markedly. CONCLUSION: The levels of IL-1 beta and IL-6 mRNA in hippocampal neurons increased with aging. GRS exerted inhibitory effects on IL-1 beta and IL-6 gene expression in chronic inflammation model of aged rats. PMID- 11501044 TI - Tripterygium wilfordii saponins and interleukin-10 prevent induction of experimental autoimmune thyroiditis by dendritic cells. AB - AIM: To study the roles of Tripterygium wilfordii saponins (TII) and interleukin 10 (IL-10) on dendritic cells (DC)-induced experimental autoimmune thyroiditis (EAT). METHODS: We used mice as autoimmune thyroiditis model animals and divided them into 4 groups, namely DC group, TII group, IL-10 group, and control group. The level of thyroglobulin (Tg) antibody was assayed by ELISA. TNF beta production in the cultured supernatants and nitric oxide (NO) in the serum were measured by biological activation assay and Griess reaction, respectively. Tg stimulated proliferation of lymphocytes was detected with MTT incorporation assay. The histopathological analysis of thyroid was carried out. RESULTS: Tg pulsed DC were able to induce EAT with increase in the concentration of TgAb in serum and lymphocytes infiltration in thyroid. After treatment with TII or IL-10, DC could not induce EAT with lower levels of TgAb and no lymphocyte infiltration. The concentration of NO in serum, TNF beta activation, and the proliferation of lymphocytes in response to thyroglobulin in TII or IL-10 group were lower than those in DC group. CONCLUSION: TII and IL-10 are able to strongly inhibit the ability of DC to induce experimental autoimmune thyroiditis. PMID- 11501045 TI - Influence of tramadol on morphine discriminative behavior in rats. AB - AIM: To study the potential of the psychological dependence of tramadol. METHODS: Rats were trained to discriminate 4.0 mg/kg morphine, and to discriminate 0.5 mg/kg methamphetamine (MA) from saline under a fixed-ratio (FR10) schedule of reinforcement. After they acquired the discrimination, different doses of tramadol were used to substitute for training dose of morphine and MA. Naltrexone was injected concomitantly with tramadol. RESULTS: Tramadol fully substituted morphine at a dose of 32 mg/kg or higher. The ED50 value of the discriminative effects of tramadol was 20.94 mg/kg, higher than that of morphine (2.04 mg/kg, P < 0.01). MA failed to generalize to tramadol at the doses tested. Naltrexone antagonized the discriminative response of tramadol. CONCLUSION: Tramadol can substitute for morphine in morphine discriminative rats. The discriminative stimulus effects of tramadol are mediated by a mu opioid mechanism. PMID- 11501046 TI - Effects of diltiazem on down-regulation of lymphocyte beta-adrenoceptors in patients with chronic congestive heart failure. AB - AIM: To determine whether diltiazem could reverse down-regulation of lymphocyte beta-adrenoceptor (beta-AR) in patients with chronic congestive heart failure (CHF). METHODS: Before and after the treatment with diltiazem in CHF patients, lymphocyte beta-AR density was measured with [3H]dihydroalprenolol radioligand binding assay, levels of free cytosolic calcium ([Ca2+]i) in platelets were estimated with fluorescent indicator Fura 2-AM, plasma norepinephrine (NE) levels were measured with 125I-radioimmunoassay. RESULTS: Lymphocyte beta-AR density was lower and [Ca2+]i in platelets was significantly higher in CHF patients than those in control. Plasma NE levels were higher in CHF patients than those in control. Diltiazem therapy reduced [Ca2+]i in platelets and increased lymphocyte beta-AR density in CHF patients without significant change of plasma NE concentration. CONCLUSIONS: Diltiazem partly reversed down-regulation of lymphocyte beta-AR density in CHF patients, and this effect was not related to the level of plasma NE, and might be attributed to intracellular [Ca2+]i decrease. PMID- 11501047 TI - Inhibition by agmatine on spontaneous activity of rabbit atrioventricular node cells. AB - AIM: To study the effects of agmatine on spontaneous activity of atrioventricular (AV) node and its action mechanisms. METHODS: Action potentials in AV node cells were recorded using intracellular microelectrode technique. RESULTS: Agmatine not only reduced the amplitude of action potential (APA), maximal rate of depolarization (Vmax), velocity of diastolic (phase 4) depolarization (VDD), and rate of spontaneous firing (RSF), but also prolonged 90% duration of action potential (APD90) in a concentration-dependent manner. The effects of agmatine (10 mmol/L) could be blocked completely by pretreatment with idazoxan (0.1 mmol/L), an imidazoline receptor (IR) and alpha 2-adrenergic receptor (alpha 2 AR) antagonist. Pretreatment with NG-nitro-L-arginine methyl ester (L-NAME, 0.5 mmol/L), a nitric oxide (NO) synthase inhibitor, did not affect the effects of agmatine on AV node cells. Elevation of Ca2+ concentration (5 mmol/L) in perfusate antagonized the effects of agmatine (10 mmol/L). Lemakalim (30 mumol/L), an ATP-sensitive potassium channel opener, inhibited the prolonging effects of agmatine on repolarization. CONCLUSION: The inhibitory effects of agmatine on spontaneous activity of AV node cells in rabbits were likely mediated by IR and/or alpha 2-AR, and were related to the reduction in calcium influx and potassium efflux. PMID- 11501048 TI - 1-Methyl-3-isobutylxanthine delays apoptosis induced by deprivation of growth factors in vascular endothelial cells. AB - AIM: To investigate the effects of 1-methyl-3-isobutylxanthine (MIBX) on apoptosis induced by deprivation of acidic fibroblast growth factor (aFGF) and serum in vascular endothelial cells (VEC). METHODS: Nuclear fragmentation was observed by fluorescence microscopy. Viability was determined by counting the cells that attached to dishes after treatments. DNA fragmentation was measured by agarose gel electrophoresis. RESULTS: The cells deprived of aFGF and serum were treated with MIBX 25-200 mumol/L for 3, 6, 9, and 12 h, respectively. Morphological changes including the formation of apoptotic bodies and DNA fragmentation of these cells were significantly suppressed by IBMX 50-200 mumol/L at 3 h and 6 h. But after 12-h treatment, no difference was observed between the treated and untreated cells. CONCLUSION: MIBX delays apoptosis of vascular endothelial cells induced by deprivation of aFGF and serum. PMID- 11501049 TI - Antiangiogenic effect of alpha-anordrin in vitro and in vivo. AB - AIM: To study the antiangiogenic effect of alpha-anordrin (alpha-Ano), a partial antagonist of estrogen receptor. METHODS: The in vivo inhibitory effect of alpha Ano on angiogenesis was determined by microvascular density (MVD) in tumors and the chicken chorioallantoic membrane (CAM) model. The in vitro effects of alpha Ano on proliferation, migration, and attachment of human umbilical vein endothelial cells (HUVEC) were assessed by trypan blue exclusion, wound-induced two-dimensional migration model, and their ability to adhere to type I collagen, respectively. The possible involvement of nitric oxide (NO) in alpha-Ano antiangiogenic effect was determined by measuring NO content using fluorescent assay. RESULTS: alpha-Ano significantly inhibited the MVD in Lewis lung carcinoma model and this effect was correlated with its inhibition of the tumor growth. alpha-Ano also showed an inhibitory effect on the angiogenesis of CAM with the inhibitory rate of 53% and such action of alpha-Ano could not be blocked by simultaneous administration of 17 beta-estrodiol, a typical agonist of estrogen receptor. In vitro studies showed that alpha-ANO obviously suppressed the proliferation and migration of HUVEC, but had no obvious effect on the attachment of HUVEC to the type I collagen. Moreover, alpha-Ano significantly reduced the level of NO released by HUVEC in a dose- and time-dependent manner. CONCLUSION: alpha-Ano possesses an antiangiogenic effect, and this effect is mediated, at least in part, by reducing the NO content and subsequently inhibiting the proliferation and migration of endothelial cells. PMID- 11501050 TI - Nociceptin inhibits electric field stimulation-induced cholinergic constrictions in rat airways. AB - AIM: To study the effect of nociceptin (orphanin FQ), a newly discovered heptadecapeptide, on cholinergic constrictions in isolated trachea and bronchus of rat. METHODS: The electric field stimulation (EFS) induced a monophasic constriction, which was due to an activation of the cholinergic nerves. RESULTS: Nociceptin 0.001-0.1 mumol/L inhibited cholinergic constriction in a concentration-dependent manner. IC50 (95% of confidence limits) were 0.06 (0.04 0.08) mumol/L and 0.07 (0.05-0.1) mumol/L in tracheae and bronchi respectively. The constrictions inhibited by nociceptin 0.01 mumol/L in traheae and bronchi were (58 +/- 32)% and (60 +/- 26)% respectively compared with the control, in which nociceptin was not applied. After pretreatment with naloxone 0.1 mumol/L, the constrictions were (60 +/- 19)% and (54 +/- 20)% (P > 0.05 vs the above figures). However, the constrictions induced by exogenous acetylcholine were unaffected by nociceptin 0.01 mumol/L. kappa-Opioid receptor agonist, U-50488H (0.01-1 mumol/L) did not affect the EFS-induced cholinergic constrictions in rat airways. CONCLUSION: Nociceptin inhibits EFS-induced cholinergic constriction, which is not affected by naloxone in rat airways. PMID- 11501051 TI - In vitro inhibition of rat monoamine oxidase by liquiritigenin and isoliquiritigenin isolated from Sinofranchetia chinensis. AB - AIM: To study the inhibition of liquiritigenin (1) and isoliquiritigenin (2) isolated from Sinofranchetia chinensis on rat monoamine oxidase A and B (MAO A and B). METHODS: Rat brain mitochondrial fraction, prepared by differential centrifugation, was utilized as a source of MAO activity. MAO activity was determined radiochemically with [14C]5-hydroxytryptamine (5-HT) and [14C]beta phenylethylamine (beta-PEA) used as MAO A or B specific radiolabled substrates, respectively. The Ki and KI values were obtained from Lineweaver-Burk plot using linear regression analysis. RESULTS: Liquiritigenin and isoliquiritigenin were found to be inhibitory against both MAO A and B in a dose-dependent manner. IC50 (95% of confidence limits) of liquiritigenin and isoliquiritigenin were 32 (26 36) and 13.9 (12.8-15.6) mumol/L for the inhibition of MAO A, and 104.6 (89.0 118.9) and 47.2 (39.5-54.5) mumol/L for that of MAO B, respectively. Lineweaver Burk transformation of the MAO A inhibition data indicated that the inhibition was non-competitive for both liquiritigenin and isoliquiritigenin whereas their inhibition of MAO B was of mixed type. Regarding MAO A inhibition, the Ki values of liquiritigenin and isoliquiritigenin were 31.5 mumol/L and 14.3 mumol/L, respectively. As to the inhibition of MAO B, the Ki and KI data for liquiritigenin were 164.7 and 15.2 mumol/L, and those for isoliquiritigenin were 62.2 and 9.3 mumol/L, respectively. CONCLUSION: Liquiritigenin and isoliquiritigen inhibited the activity of MAO A and B in rat brain mitochondria, and the latter was more active than the former. PMID- 11501052 TI - Population pharmacokinetic analysis of amikacin and validation on neonates using Monte Carlo method. AB - AIM: To make programs for population pharmacokinetic analysis and to assess the ability of this method in pharmacokinetic parameter estimation and in the prediction of serum concentrations. METHODS: Data of amikacin as a model drug were collected from 42 neonates with 142 serum samples. A one-compartment open model was used to describe the kinetics of amikacin after the intravenous infusion. Following Sheiner's idea of population pharmacokinetics, we made the programs to evaluate population parameter and individual parameter. The target function minimality was obtained from Monte Carlo algorithm. The validation of the population analysis was performed using classic pharmacokinetic program 3p87 for antithesis. The predictability of the developed method was evaluated by computing precision and accuracy of serum concentration predicted using the parameter estimates. RESULTS: The stability of our self-made program was good. The population parameters obtained from this approach were in conformity with those from 3p87, and the interindividual variability was relatively small. For the learning sample and the validation sample, predicted and observed concentrations were all close with correlation coefficient 0.995 and 0.990, respectively. Most of predicted errors were found < +/- 1 mg/L, and RMSD and BIAS were 0.58 and -0.07 for the validation sample, respectively. The choice of blood sampling time was an important factor for the predictive performance. An early sampling time after the infusion was observed to be the best sampling time. CONCLUSION: The estimation program of population parameter and individual parameter made by us ran stably, and allowed us to use sparse data to estimate population pharmacokinetic parameters. It provided accurate estimates of these parameters and satisfactory ability of serum concentration prediction. Therefore, it can be used for the population pharmacokinetic analysis and individualization of dosage regimen. PMID- 11501053 TI - Sympathectomy induces novel purinergic sensitivity in sciatic afferents. AB - AIM: To test the hypothesis that sympathectomy could induce novel purinergic sensitivity in sciatic afferents. METHODS: Teased-fiber recordings were made from 32 spontaneously active A afferents from the sciatic nerves in surgically sympathectomized rats and 30 spontaneously active A afferents from the sciatic nerves in intact rats. Adenosine 5'-triphosphate (ATP) was injected via a cannula in jugular vein. RESULTS: Twenty eight percent of the spontaneously active afferent fibers from sciatic nerves in the sympathectomized rats responded to ATP, either with an increase or with a decrease in spontaneous firing. However, none of the fibers from the sciatic nerves in the intact rats was activated by ATP. CONCLUSION: Sympathectomy induces novel purinergic sensitivity in A afferents from sciatic nerve. PMID- 11501054 TI - Adenovirus-mediated expression of antisense RNA transcripts complementary to pig alpha(1,3) galactosyltransferase mRNA inhibits expression of Gal alpha(1,3) Gal epitope. AB - AIM: To examine the effects of the expression of antisense RNA transcripts complementary to the pig alpha(1,3) galactosyltransferase [alpha(1,3)GT]mRNA on the expression of Gal alpha(1,3) Gal structure (gal epitope) in cultured cell lines. METHODS: Human adenoviral vectors were used to mediate the expression of antisense RNA. The expression levels of H blood group antigens and gal epitopes were analyzed by flow cytometry using FITC-UEA-I and FITC-GS-IB4 lectins, respectively. RESULTS: Recombinant adenoviruses, Ad5anti-sGT600 and Ad5-anti sGT1100, which express antisense RNA complementary to different regions of the pig alpha(1,3) GT mRNA, were constructed and used to infect cell line of NIH3T3. The results showed about 30% reduction in the expression level of gal epitopes on the surface of NIH3T3 cells. In addition, co-expression of human secretor type alpha(1,2) fucosyltransferase [alpha(1,2)FT]cDNA and antisense RNA complementary to the pig alpha(1,3) GT mRNA led to a further reduction in the gal epitope level. CONCLUSION: Recombinant adenoviruses, Ad5anti-sGT600 and Ad5anti-sGT1100, are effective to down-regulate the gal epitope expression. PMID- 11501055 TI - Influence of idazoxan on analgesia, tolerance, and physical dependence of morphine in mice and rats in vivo. AB - AIM: To study the influence of idazoxan (Ida), an antagonist of imidazoline receptors (I-R), on analgesia, tolerance, and physical dependence of morphine. METHODS: The effects of Ida on pain threshold and morphine analgesia were observed in mouse acetic acid writhing test and 55 degrees C hot plate test. The effect of Ida on morphine tolerance and physical dependence were observed in mouse tolerant model and in mice and rat models. RESULTS: Ida (3-9 mg/kg) significantly decreased the pain threshold by 120% in acetic acid writhing test and by 39% in 55 degrees C hot plate test of mice, respectively. It inhibited the analgesic effect of morphine in a dose-dependent manner. Ida promoted the development of tolerance to morphine in mice and induced the abstinence syndrome in morphine-dependent mice and rats similar to naloxone. CONCLUSION: I-R and its endogenous ligand agmatine might participate in the pain threshold and influence morphine analgesia as well as negatively regulate tolerance to and physical dependence on morphine. PMID- 11501056 TI - Blocking effect of tricyclopinate on nicotinic receptors in cultured sympathetic neurons. AB - AIM: To investigate the mechanism of tricyclopinate, an antagonist of nicotinic receptor, on neuronal nicotinic acetylcholine receptors (nAChR). METHODS: A tight seal whole-cell recording patch-clamp technique was performed to record nicotine evoked currents in the cultured sympathetic neurons from neonatal rat superior cervical ganglia (SCG). RESULTS: Tricyclopinate inhibited the nicotine-induced currents competitively and the inhibition was voltage-independent. The decay of the nicotine-induced current was accelerated significantly in the presence of tricyclopinate. CONCLUSION: Tricyclopinate inhibits neuronal nAChR by interacting with the allosteric sites rather than the open ionic channels or acetylcholine recognition site of the receptor. PMID- 11501057 TI - High-throughput screening for human collagenase 1 inhibitors. AB - AIM: To establish a high-throughput method for inhibitor screening using a recombinant collagenase catalytic domain. METHODS: Human collagenase 1 catalytic domain protein was expressed in E coli and used for screening a set of 2720 compounds in a high-throughput fashion. RESULTS: The screening was accomplished within 2 h and 10 min with consumption of each compound at 4 micrograms. Sixty six compounds were identified with > 60% inhibitory activity at 20 mg/L, among which 44 compounds were confirmed by subsequent testing at multiple concentrations. The most potent compound showed an IC50 at 4.3 mumol/L, and there were total 15 compounds with IC50 less than 20 mumol/L. CONCLUSION: The high throughput method using the recombinant collagenase is fast, effective and practical in identifying inhibitors. PMID- 11501058 TI - Determination of fluoxetine and its metabolite norfluoxetine in human liver microsomes by reversed-phase HPLC in vitro. AB - AIM: A high-performance liquid chromatography (HPLC) method was developed for the determination of fluoxetine (FLU) and its metabolite norfluoxetine (N-FLU) in human liver microsomes in vitro. METHODS: An incubation buffer containing human liver microsomes, NADPH-generating system, and FLU, after termination of enzyme reaction and addition of nortriptyline (NOR) as internal standard (IS), was extracted with n-hexane/acetonitrile, and separated on a reversed-phase ODS column. Detection was achieved at 226 nm by ultraviolet detector (UV). RESULTS: The limit of detection was 5 micrograms/L for both FLU and N-FLU. No potential interference was found. The method provides recoveries of up to 94%-104% and acceptable coefficients of variation were found for both within-run (< 7.8%) and day to day (< 9.1%) assays. CONCLUSION: This method is rapid, sensitive, and simple for studying the metabolism of FLU and N-FLU. PMID- 11501059 TI - G-2964A and C734A genetic polymorphisms of CYP1A2 in Chinese population. AB - AIM: To observe the G-2964A and C734A genetic polymorphisms of human CYP1A2 in Chinese population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed in genotyping analysis. RESULTS: The allele frequencies of A-2964 were 0.25 and 0.22 in Qidong and Changsha populations, respectively. The incidences of A734 were 0.68 in Qidong population and 0.66 in Changsha population. No more than two low-inducibility/activity alleles were presented in one person. CONCLUSION: The distribution of the G-2964A and C734A genetic polymorphisms did not show significant difference between Chinese and Japanese populations. The incidence of C734A in Chinese was also similar to that in Caucasian population. PMID- 11501060 TI - Effects of angiotensin II on release of CRH and AVP from hypothalamus during acute hypoxia. AB - AIM: To investigate the effects of angiotensin II (Ang II) on corticotropin releasing hormone (CRH) and argipressin (AVP) release from median eminence (ME) of hypothalamus during acute hypoxia in rats. METHODS: Simulated hypoxia was performed in a hypobaric chamber. CRH and AVP were determined by radioimmunoassay (RIA). Plasma corticosterone concentration was measured by fluorometry. RESULTS: Ang II did not influence CRH release induced by hypoxia [CRH in group pretreated with Ang II (16 +/- 8) ng/ME vs control (15 +/- 4) ng/ME, both exposed to hypoxia with simulated altitude 7 km (8.2% O2)]. Ang II enhanced AVP release, from (5.7 +/- 1.6) ng/ME in control decreasing to (2.6 +/- 1.2) ng/ME (P < 0.05), meanwhile plasma corticosterone concentration was also increased markedly, from (356 +/- 58) in control to (536 +/- 134) micrograms/L plasma (P < 0.05), which was partly abolished by administration of AVP antiserum. CONCLUSION: Ang II might stimulate hypothalamo-pituitary-adrenal axis (HPA) through activating AVP but not CRH release during acute hypoxia. PMID- 11501061 TI - Plasma CPU-86017 concentrations regarding suppression of ouabain-induced cardiac arrhythmias and decrease of heart rate in guinea pigs. AB - AIM: To determine the effective plasma levels of CPU-86017 which could suppress the cardiac arrhythmias induced by i.v. ouabain in guinea pigs. METHODS: The cardiac arrhythmias and the heart rate were monitored by ECG traces. Blood samples were collected to determine plasma levels using HPLC assay. TXB2 and 6 keto-PGF1 alpha were measured in plasma. RESULTS: The plasma concentrations of CPU-86017 which were effective to suppress ventricular fibrillation (VF) and heart rate were 0.13-0.23 mg/L and 0.13-0.31 mg/L, respectively. A reduction of TXB2 levels and an elevation of 6-keto-PGF1 alpha levels were observed after CPU 86017 i.v. administration. CONCLUSION: The arrhythmia-suppressing and heart rate slowing effect of CPU-86017 followed a linear relationship with its concentrations in plasma. PMID- 11501062 TI - Effect of combination of valsartan with benazepril on blood pressure and left ventricular hypertrophy in SHR. AB - AIM: To evaluate the antihypertensive efficacy of angiotensin converting enzyme inhibitor (ACEI) benazepril in combination with AT1 receptor antagonist valsartan and their effect on left ventricular hypertrophy, renin angiotensin aldosterone system (RAAS) and endoxin in spontaneously hypertensive rat (SHR). METHODS: WKY control group (n = 6) and other 4 groups consisted of 24 SHR (14-week-old, male, n = 6): SHR control group, benazepril group, valsartan group, and combination drug therapy group. Systolic blood pressure (SBP) of SHR was measured at the beginning and at the end of 2, 4, 6, and 8 wk of drug intervention. Morphometric determination, renin activities, angiotensin II (Ang II), endoxin, and ATPase activity analysis were performed at the end of 8 week of drug intervention. RESULTS: SBP, ratio of left ventricular mass (LVM), body weight (BW) (LVM/BW), and transverse diameter of myocardial cell (TDM) of SHR were remarkably decreased after drug intervention, and this decrease was most remarkable in the combination drug therapy group. Renin activities of plasma and myocardium were remarkably increased in drug intervention groups. The levels of Ang II in plasma and myocardium were remarkably increased in valsartan group, decreased in benazepril group and combination drug therapy group. Na(+)-K(+)-ATPase activities in myocardium were remarkably increased and the level of endoxin in myocardium were remarkably decreased as SBP decreased after drug intervention. CONCLUSION: Both benazepril and valsartan can decrease SBP of SHR, and cause regression of ventricular hypertrophy. The efficacy of combination drug therapy group is most remarkable among all groups and avoids the side effects of induction of high Ang II levels in plasma and myocardium caused by long-term use of valsartan alone. PMID- 11501063 TI - Benznidazole-induced ultrastructural and biochemical alterations in rat colon. AB - AIM: To study the effects of benznidazole (Bz), a drug used in the chemotherapy of the acute and the intermediate phase of Chagas' disease, on the colon of rats. METHODS: Sprague Dawley male rats received Bz 100 mg/kg ig. After 24 h colons were examined by electron microscopy. Concentrations of Bz in colonic tissue were measured by HPLC. Bz nitroreduction was followed by the decrease in the drug concentration using spectrophotometry and HPLC or by covalent binding to proteins of reactive products formed under in vivo and in vitro conditions. RESULTS: Colon mucosa of Bz-treated rats showed intense ultrastructural alterations: abundant mucus secretion at the level of the Goblet cells and dilatation of the endoplasmic reticulum and the Golgi apparatus in epithelial cells. The concentration of Bz in tissue was (59 +/- 18) and (93 +/- 14) nmol/g (protein) 1 and 3 h after oral administration to rats, respectively. Colonic microsomes anaerobically activated Bz in the presence of NADPH. This activating nitroreductive pathway only involved a minor part of the total Bz and could not be detected spectrophotometrically or by HPLC analysis of the Bz consumed. Reactive metabolites that bound covalently to microsomal proteins were formed in this process. The covalent binding was also observed in vivo 1, 3, 6, and 24 h after administration of the labeled drug to rats. CONCLUSION: Reactive Bz metabolites produced during nitroreductive bioactivation of the drug in the colonic mucosa could interact with proteins and other cellular constituents to cause injury. PMID- 11501064 TI - Effect of selective IL-6 inhibitor Am-80 on experimental autoimmune encephalomyelitis in DA rats. AB - AIM: To observe the role of interleukin (IL)-6 in the development of experimental autoimmune encephalomyelitis (EAE). METHODS: DA rats were immunized by injecting bovine myelin basic protein (MBP). mRNA of cytokines, such as IL-6, IL-10, TNF alpha, TGF-beta 1, IFN-gamma, and iNOS, were detected by RT-PCR. MBP was injected into ear to induce delayed type cutaneous hypersensitivity response (DTH). Histological studies were performed on the spinal cord with HE staining. Nitric oxide (NO) production from cultured murine macrophage clones was stimulated with LPS plus IFN-gamma. RESULTS: DA rats developed EAE disease with a peak of severity on d 13 and d 14. Am-80 (1.0, 3.0 mg/kg), a selective IL-6 inhibitor, inhibited the symptoms in terms of deterioration as observed by the clinical score, body weight and histological findings, in a dose-related manner. A high dose of Am-80 (3.0 mg/kg for 12 d) did not completely inhibit the disease, but delayed the symptoms and enhanced the delayed response. By prolonging the duration of treatment (18 d), Am-80 inhibited the onset of EAE during administration, but the symptoms of EAE appeared after the administration was stopped. Am-80 administerd for 12 d inhibited the DTH response on d 11 but not on d 22. RT-PCR studies demonstrated a strong expression of IFN-gamma, IL-6, IL-10, TGF-beta 1, TNF-alpha, and iNOS mRNA in spinal cord 13 d after immunization. However IFN-gamma, IL-10, TNF-alpha, and iNOS mRNA expression (on d 13) was suppressed by Am-80, except in the case of IL-6, hence the effect of Am-80 on the expression of IL-6 mRNA was examined in additional experiments. After Am-80 was administered for 12 d or 18 d, the expression of IL-6 mRNA was inhibited on d 12 or d 18, but increased on d 13 or d 19, respectively. CONCLUSION: These findings suggest that inhibition of EAE by Am-80 is initiated by inhibition of IL-6 production. PMID- 11501065 TI - Extracellular detection of delayed afterdepolarization of cardiac fibers using signal averaging technique. AB - AIM: To detect delayed after depolarizations (DAD) in extracellular electrograms using signal averaging technique. METHODS: DAD were induced by acetylstrophanthidin (0.25 mumol.L-1, n = 9) and resibufogenin (0.52 mumol.L-1, n = 5) in sheep cardiac Purkinje fibers. Intracellular voltage was recorded with a conventional microelectrode, and simultaneous extracellular electrograms were recorded differentially from widely spaced electrodes placed in the tissue bath. Noise of electrograms was reduced using signal averaging technique. RESULTS: Acetylstrophanthidin and resibufogenin both induced DAD in the intracellular recording and extracellular DAD (DAD-E) in the extracellular electrogram in sheep heart Purkinje fibers. Acetylstrophanthidin and resibufogenin induced typical changes in the action potential including decrease in action potential amplitude, resting potential, maximum diastolic potential, and action potential duration. Similar shortening occurred in the "Q-T interval" recorded by the extracellular electrogram. With either acetylstrophanthidin or resibufogenin, shortening of stimulation cycle length from 990 ms to 690 ms reduced the coupling interval between action potential upstroke and peak voltage of the DAD (P < 0.01), and the coupling interval between the "QRS" and DAD-E recorded extracellularly (P < 0.01). CONCLUSION: DAD can be detected using the extracellular electrograms combined with the high resolution, signal averaging technique. PMID- 11501066 TI - Swelling-activated chloride currents in embryonic chick heart cells. AB - AIM: To characterize a swelling-activated chloride current, I(Cl, swell), in white Leghorn chick heart cells and the effects of chlorpromazine (CPZ) effects. METHODS: The patch-clamp technique in the whole-cell configuration was used. RESULTS: Hyposmotic swelling elicited I(Cl, swell) in white Leghorn chick heart cells. The current amplitude increased from (452 +/- 200) pA to (849 +/- 373) pA with a reduction of osmolarity from 300 mmol.L-1 to 270 mmol.L-1, 4',4 Diisothiocyanostilbene-2,2'-disulphonic acid (DIDS) 100 mumol.L-1 decreased I(Cl, swell) from (1196 +/- 505) pA to (830 +/- 328) pA in hyposmotic solution. In white Leghorn chick heart cells I(Cl, swell) was not induced by CPZ 30 mumol.L-1, which is different from the case of E coli spheroplast. CONCLUSION: Swelling activated chloride current was activated by hyposmotic swelling in white Leghorn chick heart cells. The mechanism for activating the current is different from that of mechanosensitive ion channels of E coli. PMID- 11501067 TI - Effects of p38 and p42/p44 CCDPK signaling on H2O2-induced apoptosis in bovine aortic endothelial cells. AB - AIM: To investigate the effects of p38 and p42/p44 Ca(2+)-calmodulin dependent protein kinases (CCDPK) signaling on hydroperoxide (H2O2)-induced apoptosis in cultured bovine aortic endothelial cells (BAEC). METHODS: Morphologic changes and quantification of apoptotic cells were determined under fluorescence microscope after a 24-h treatment of BAEC by H2O2. Cell viability was determined with MTT method. DNA fragmentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 and phospho-p42/p44 CCDPK was measured by Western blotting. RESULTS: H2O2 elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation) and DNA fragmentation. At 100-500 mumol.L-1, incubation of BAEC with H2O2 for 24 h also induced phospho-p38 and phospho p42/p44 CCDPK expression in a concentration-dependent manner. Interestingly, H2O2 induced apoptosis was markedly increased by preincubation with U0126, a specific p42/p44 CCDPK inhibitor. However, SB203580, a specific p38 CCDPK inhibitor, enhanced the expression of phospho-p42/p44 CCDPK induced by H2O2, but had no effect on BAEC survival. CONCLUSION: p42/p44 CCDPK signaling appears to play protective roles in H2O2-induced apoptosis in BAEC, whereas p38 CCDPK is not the main signaling pathway mediating H2O2-induced cellular apoptosis. PMID- 11501068 TI - FUT2 gene involved in expression of H blood group antigen on surface of human tumor cell lines BEL-7404, SGC-7901, and SPC-A-1. AB - AIM: To investigate the expression of blood group H antigen on the surface of several human tumor cell lines such as BEL-7404, SPC-A-1, and SGC-7901. METHODS: In vitro and in vivo expression of the blood group H antigen was analyzed with flow cytometry and immunohistochemistry. Expression of FUT2 gene was determined by RT-PCR, Southern blot, and restriction digestion. RESULTS: Flow cytometric analysis showed that SGC-7901 cells, BEL-7404 cells, and SPC-A-1 cells had mean fluorescence intensity (MFI) values of 162 +/- 43, 81 +/- 25, and 28 +/- 17, respectively. DNA fragments of about 1.0 kb in length were obtained by RT-PCR from RNA isolated from these cells and were detected by FUT2-specific [alpha 32P]dATP labeled DNA probes. CONCLUSION: Human tumor cell lines BEL-7404, SGC 7901, and SPC-A-1 all expressed blood group H antigen on their cell membranes in vitro and in vivo, but their expression levels varied significantly between different cell lines. FUT2 gene expression resulted in the production of these antigens on the cell membrane. PMID- 11501069 TI - [Relevant problems in the management of inhalation injury]. PMID- 11501070 TI - [The changes in respiratory cycle, ventilation volume and blood analysis after bronchoalveolar lavage in dogs with steam inhalation injury]. AB - OBJECTIVE: The changes in respiration cycle, ventilation volume and blood analysis after bronchoalveolar lavage(BAL) in dogs with steam induced inhalation injury were investigated to explore ways of treating inhalation injury. METHODS: A bronchoscope was introduced down to the openings of the tertiary-quarternary bronchioles, and controlled volume BAL with drugs was done. RESULTS: The inspiration time (Ti) and respiration cycle (Te) at 30, 60, 90 minutes after BAL were significantly lower than those of pre-BAL(P < 0.05), and respiration rate (RR) post-BAL at the same time points was markedly higher than that of pre-BAL(P < 0.05 or P < 0.01). The inspiratory minute ventilation volume (Vi) and expiratory minute ventilation volume (Ve) at 30 to 60 minutes after BAL were obviously increased as compared to pre-BAL(P < 0.01-P < 0.05); Vi and Ve at 90 minutes after BAL were also higher than those of pre-BAL, but there was no significant difference between them. The PaO2 at 30 minutes after BAL declined obviously as compared to pre-BAL(P < 0.05), and then rose gradually to approach pre-BAL level compared with pre-BAL. No significant changes were found in the expiration time (Te), ration of Ti to Te (Ti/Te), inspiratory tidal volume (Vti), expiratory tidal volume (Vte), PaCO2 and pH between values before and after BAL at all time points. CONCLUSION: Controlled volume BAL with mixed drugs could improve lung alveolar ventilation, and it might have an excitory effect on respiration with potential clinical value. PMID- 11501071 TI - [Experience in the management of severe inhalation injury]. AB - OBJECTIVE: To summarize our clinical experiences in the treatment of severe inhalation injury accompanying extensive burn. METHODS: Retrospective analysis of 30 cases of inhalation injury (1980-1996) was done, the cure rate, mortality, effective treatments and lessons of failure were summarized. RESULTS: As soon as the diagnosis was defined, tracheostomy should be done immediately, followed by oxygen therapy, tracheo-bronchial lavage, aspiration, and clearance of airway secretion. If there was pulmonary edema or severe bronchospasm, cortical hormone was used for a short time, in addition, the patients were encouraged to cough, breathe deeply, and change postures, as well as patted on the back and also positioned to facilitate posture drainage, etc. These measures were effective. In this series, there were thirty cases of severe inhalation injury. Fourteen patients(46.6%) were cured, sixteen patients died(53.3%). The result was basically satisfactory. CONCLUSION: It is possible to increase the cure rate of severe inhalation injury, and the treatment should be carried out conscientiously as early as possible, and great attention should be paid to prevent various complications. PMID- 11501072 TI - [Analysis of the treatment of 82 patients with inhalation injury]. AB - OBJECTIVE: To learn the experiences and improve the treatment of inhalation injury. METHODS: Causes of death and treatment effects were analyzed in a group of 82 patients admitted from January 1994 to April 1998. RESULTS: Among the 82 patients, 33 patients were cured, 11 patients were improved and 38 patients died. Among the 38 cases of death, 24 patients died directly of inhalation injury and 14 patients died of other complications. CONCLUSION: The major causes of death of the patients with inhalation injury are acute intoxication, asphyxia, and ARDS(acute respiratory distress syndrome). For the treatment, tracheostomy should be carried out the earlier the better. Moistening of the airway and lavage may be effective in preventing lung infection, facilitating the ejection of phlegm and necrotic mucosa. The amount of fluid infusion should appropriately be increased and this will not lead to lung edema. Administration of corticol hormone in the early stage for a short period will also result in positive effects. PMID- 11501073 TI - [Clinical experiences in the treatment of 77 cases of inhalation injury]. PMID- 11501074 TI - [Surveillance and diagnosis of laryngeal burn]. AB - OBJECTIVE: To explore early diagnosis and treatment of laryngeal burn, in order to prevent laryngeal obstruction, define the prevention of laryngemphraxis, and elect optimal time of treatment and appropriate treatment. METHODS: A total of 18 patients suffering from laryngeal burn were repeatedly examined with a fiberlaryngoscope. Each part of the larynx was assessed for the degree of congestion, swelling, blister formation and impairment of laryngeal function, and the effects of these pathologies on breathing. RESULTS: Our observations showed that classification of inhalation injury according to anatomical division could not reflect the severity of the injury. For this reason, every changes in each part of the larynx was analyzed. A classification consisting of 3 types of laryngeal injury was proposed in respect to 3 predisposing factors of laryngeal obstruction. This classification could serve as a guide for treatment. CONCLUSION: A clearcut diagnosis of laryngeal inhalation injury is a pre requisite for the choice of the correct treatment and the decision on the time for surgical intervention, thus laryngeal obstruction could be avoided, and death rate of early stage of inhalation injury could also be lowered. PMID- 11501075 TI - [The role and significance of PAF in the early pulmonary injury following burns combined with endotoxemia in rats]. AB - OBJECTIVE: To investigate the role and significance of PAF in the early pulmonary injury following burns combined with endotoxemia in rats. METHODS: An animal model, in which rats were subjected to 20% TBSA III degree burns combined with intraperitoneal injection of lipopolysaccharide (LPS), was used for this experiment. The changes in the pulmonary morphology were observed by LM, EM and immunohistochemistry(IHC). The contents of PAF and TNF-alpha in the plasma were determined with quantitative analysis. PAF receptor antagonist(BN50739) was used to treat early pulmonary injury. RESULTS: PAF was one of the inflammatory mediators to precede TNF changes during burns combined with endotoxemia in the early stage of pulmonary injury. The contents of PAF in the plasma was positively correlated to the extent of pulmonary injury, which may be alleviated after injection of PAF receptor antagonist (BN50739). CONCLUSION: The results suggest that PAF plays an very important pathogenic role in the early lung injury resulted from burns combined with endotoxemia. PMID- 11501076 TI - [The relationship between tissue tumor necrosis factor-alpha(TNF-alpha) mRNA expression and gut-derived endotoxemia in rats after thermal injury]. AB - OBJECTIVE: To investigate the dynamic changes in tumor necrosis factor-alpha(TNF alpha) mRNA expression and the relationship between TNF-alpha mRNA and gut derived endotoxemia in rats after thermal injury. METHODS: 56 male Wistar rats were subjected to a 35% total body surface area full-thickness thermal injury and randomly divided into 3 groups: normal controls, 35% III* injury group, and polymyxin B treatment group. Plasma endotoxin was measured by limulus amebocyte lysate test, and tissue TNF-alpha mRNA expression was determined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: It was found that a significant elevation of endotoxin concentration in both portal and systemic circulation was observed at 2 hours after thermal injury, peaking at 8 hours (P < 0.05-0.01) and lowering at 24 hours. Meanwhile, TNF-alpha mRNA expression significantly increased in liver, lung, intestine, and kidney at 2 hours, peaking at 8 hours, and a high level was maintained till 24 hours. However, treatment with a low-dose polymyxin B could lower both portal and systemic endotoxin levels, and markedly inhibit TNF-alpha mRNA expression in various organs. In addition, a positive correlation between portal endotoxin concentration and pulmonary, intestinal and renal TNF-alpha mRNA expression was noted (r = 0.365 0.484, all P < 0.05). CONCLUSION: This study suggests that major burns can lead to a marked elevation of tissue TNF-alpha mRNA expression in vital organs, which may be associated with gut-derived endotoxemia caused by acute thermal injury. PMID- 11501077 TI - [Effects of nitric oxide on platelet function in burned rats]. AB - OBJECTIVE: To investigate the effects of nitric oxide (NO) on rat platelet function after burn injury. METHODS: Wistar rats with 30% TBSA full thickness burns were used as model, indices of platelet functions including platelet aggregation rate in first minute (PAR-1), maximal platelet aggregation rate (PAR m), platelet dissipation rate (PDR) and platelet adhesion rate (PAR) were determined in four groups, namely burn control (B), NO donor (sin-1), NO synthesis inhibitor (L-NAME) and the normal control (C). RESULTS: The PAR-1, PAR m and PAR were significantly increased and the PDR was decreased remarkably after burn injury. In sin-1 group the PAR-1, PAR-m and PAR were obviously lower and the PDR was higher than that of B group (P < 0.05-0.01). In L-NAME group the PAR-1 was higher and the PDR was lower as compared with B group, but the PAR-m and PAR showed no significant differences between two groups. CONCLUSIONS: The ability of platelet adhesion and aggregation induced by activated platelet are increased after burn injury. NO can partially reduce the degree of platelet activation and inhibit platelet aggregation. Moreover, it may dissipate aggregated platelets. PMID- 11501078 TI - [Morphological study of CD3+ dendritic epidermal T cells in human skin and scar]. AB - OBJECTIVE: To investigate the CD3/TCR-bearing T cell in the epidermis and explore the influence of the dendritic epidermal T cells on regeneration and differentiation of epidermal cells. METHODS: The CD3/TCR-bearing T cells in epidermis of human skin and cicatrix were examined by means of histomorphology and immunohistochemistry. RESULTS: It was found that the density of CD3/TCR bearing T cells in human skin and scar tissue had great dissimilarity. CD3+ dendritic epidermal T cells increased in hyperplastic scar. Such result was not observed in atrophic scar. CONCLUSION: Our findings indicate CD3+ dendritic epidermal T cells regulate the generation and differentiation of epidermal cells. CD3+ dendritic epidermal T cells may play an important role in the maintenance of the morphology of epidermal tissue. PMID- 11501079 TI - [The effects of IL-1 on synthesis of collagen in scar-derived fibroblasts]. AB - OBJECTIVE: To investigate modulation of cytokines on biological activities of scar-derived fibroblasts. METHODS: This study focused on the effects of interleukin-1 (IL-1) on collagen biosynthesis, nucleic acid, and cell organs of fibroblasts. RESULTS: The results showed the IL-1 could reduce not only the collagen synthesis of fibroblasts, but also quantity of nucleic acid. CONCLUSION: The authors confirm that IL-1 is a cytokine with down-regulation effect on scar formation, and suggest that IL-1 can play an important role in prevention and treatment of scar. PMID- 11501080 TI - [The architecture of the cutaneous blood vessels in scent pigs]. AB - OBJECTIVE: To study the anatomy of cutaneous blood vessels of the scent pig, and to provide an animal model for research in plastic surgery. METHODS: Seven Chinese mini-pigs (scent pigs) were used in the study. RESULTS: 1. The scent pig is small, easy to house, rich in panniculus adiposus. 2. The origin of cutaneous blood vessels is divided into three types: the musculo-cutaneous perforator, the intermuscular septal perforator and the direct cutaneous vessel. There are some differences in distribution, size, and architecture from the characteristics of that of human being. 3. The architecture of cutaneous vessels could be divided into four vascular plexuses: the profound vascular plexus, middle vascular plexus, superficial vascular plexus of the superficial fascia and the subdermal vascular plexus. 4. The density of vessels and vessel plexuses in the neck and trunk is higher than that of the legs. CONCLUSION: In terms of the morphology of cutaneous blood vessels, the scent pig is an ideal animal model for skin flap study. PMID- 11501081 TI - [Experimental study on the relationship between the ratio of pedicle width to flap width and the surviving length of the random skin flap]. AB - OBJECTIVE: To clarify the influence of the ratio of pedicle width to flap width of a random flap on flap survival length. METHODS: 42 skin flaps in seven groups were created in 6 pigs' torso with different ratio of pedicle width to flap width. The flaps were evaluated with fluorescence examination, blood flow ECT test, and computerized analysis of survival area. RESULTS: The flap survival length of the seven groups was the same on the whole. The change in the ratio of pedicle width to flap width has nothing to do with flap survival length as long as the ratio is within the range of 1:1 to 1:4. CONCLUSION: The random flap has a fixed survival length regardless of the width of the pedicle and the flap. The survival area of a random flap seems to approach a semicircle with a radius of the fixed survival length of the flap. PMID- 11501082 TI - [Adenosine is effective to improve the viability of the transverse rectus abdominis myocutaneous (TRAM) flap in the pig]. AB - OBJECTIVE: The aim of this experiment was to investigate the adenosine treatment for augmentation of TRAM flap viability in pigs. METHODS: This TRAM flap model was based on the deep inferior epigastric vascular pedicle, with the center of the transverse skin pedicle attached to the underlying rectus abdominis muscle at the superior end of the muscle and extending bilaterally from its attached muscle. The transverse skin pedicle (6 x 30 cm) included a contralateral and ipsilateral random portion of skin. Prior to raising the TRAM flap, 1 mg, 2 mg or 5 mg adenosine was injected through the superior epigastric artery in the experiment group. RESULTS: Treatment with 2 mg or 5 mg adenosine increased skin viability of the transverse skin flap in the experiment group compared with the sham-operated control (P < 0.05, n = 5). CONCLUSION: Adenosine treatment through a dominant vascular pedicle prior to raising the TRAM flap is effective in augmenting skin viability of the flap. PMID- 11501083 TI - [MRI features of lymphedema of the lower extremity: comparison with lymphangioscintigraphy]. AB - OBJECTIVE: To investigate the features of chronic lymphedema of the extremity. METHODS: Magnetic resonance imaging (MRI) and lymphangioscintigraphy (LAS) examinations were performed on 12 patients with peripheral lymphedema. RESULTS: MRI characteristically showed diffusive subcutaneous edema, reticular lymphangiectasis and "channels" with sequestered lymph. MRI scan clearly displayed the proliferative and extended lymphatic vessels, trunks and chylocyst. LAS showed dermal diffusion (dermal backflow) or retention at the injection site of the tracer with poorly defined lymphatic trunks and delayed or no visualization of regional lymph nodes. CONCLUSION: MRI can visualise peripheral lymph trunks, lymph nodes and soft tissue. LAS is more helpful in ascertaining the condition of obstruction of lymphatic system. Using these two imaging modalities together is helpful for anatomical diagnosis and delineating the disarranged pattern of lymphedema. PMID- 11501085 TI - [Comparison of tongue position and movement pattern before and after repair in cleft palate patients]. AB - OBJECTIVE: To study the influence of tongue position and movement pattern on the phonation of cleft palate patients. METHODS: The standard lateral cephalometric roentgenographic recordings were performed at rest and functional position. Computer scanning system was used to measure and analyse the graphs. RESULTS: At rest position, A/F ratio in pre-operation group is smaller than in post-operation and control groups; C/H ratio in post-operation group is greater than in pre operation and control groups; S/V ratio in post-operation group is smaller than in control group. During phonation of /a:/, D/F, A/P, A/F and a/p ratios in post operation group are smaller, but P/B, C/H ratios are bigger than in control group. A/F ratio in post-operation group is smaller and C/H ratio is bigger than in control group. D/F, A/P, a/p ratios in pre-operation group are smaller than in post-operation group. CONCLUSION: Significant abnormalities of tongue position and movement pattern exist in pre-operation group. After operation for cleft palate, most abnormalities disappeared, but some still persist. It is necessary for the patients to have phonetic training to eliminate these abnormalities and achieve clear phonation. PMID- 11501084 TI - [The principles of use of antibiotic drugs in plastic surgery: a preliminary exploration]. AB - OBJECTIVE: To investigate the use of antibiotic drugs in the plastic surgery patients. METHODS: With a random-sampling method, the patterns of use of antibiotic drugs were reviewed in 840 cases (1014 surgical operations) from the Plastic Surgery Hospital, the Chinese Academy of Medical Sciences. RESULTS: About 99.21 per cent of patients received antibiotic drugs, and prophylaxis accounted for about 99.70 per cent of all antibiotic drugs. The drugs used most often for prophylaxis were penicillin, followed by cephalosporins. CONCLUSION: By analysing the data, it is clearly shown that there exist abuse and inappropriate use of antibiotic drugs. Surgeons must keep in mind that antibiotic prophylaxis should be used with appropriate indications, optimum time and duration. PMID- 11501086 TI - [Repair of skin defects of the hand with the reverse forearm fasciocutaneous flap on the terminal radial artery]. AB - OBJECTIVE: A large reverse forearm fasciocutaneous flap has been developed and used to repair defects of the hand. METHODS: Supplied by the terminal branches of the radial artery, the reverse forearm fasciocutaneous flap was designed as large as 3-5 cm above the elbow with its pedicle located 3-5 cm proximal to the radial styloid process. The flap can cover a large skin defect of the palm and back of the hand. RESULTS: We have successfully treated 42 cases of hand skin defects with the terminal radial artery reverse forearm fasciocutaneous flap. Postoperative follow-up in 18 cases revealed satisfactory results cosmetically and functionally. CONCLUSION: The terminal radial artery reverse fasciocutaneous flap is a simple, safe and less traumatic method for treatment of hand skin defects. PMID- 11501087 TI - [The use of reverse fasciocutaneous pedicle flap in repairing electrical burn wound on extremity]. AB - OBJECTIVE: This is to introduce a method for reconstruction of extremities with electrical injury. METHODS: Thirty-six reverse fasciocutaneous pedicle flaps were designed to be used for the reconstruction of extremities with electrical injury in 20 patients. The size of the flaps ranged from 5 cm x 3 cm to 18 cm x 8 cm. The length and width of the pedicle ranged 3-6 cm and 1.5-4 cm. The ratio of the length to width of the flap ranged from 1.6:1 to 4.5:1. All flaps had been successfully transplanted with 180 degrees rotation. RESULTS: All flaps survived, except individual flaps showing mild swelling and disperse blisters. CONCLUSION: Satisfactory results both in functional and cosmetic aspects had been achieved with this kind of flap after followed-up for over one year. PMID- 11501088 TI - [Growth factors and wound healing]. PMID- 11501089 TI - [Cleft lip repair using a lateral columellar flap]. AB - OBJECTIVE: Since the important cosmetic structures of a good looking lip, viz. the philtrum column, philtrum dimple and Cupid's bow of a cleft lip repaired with the commonly used methods, are still defective in various degrees, a new method is designed. METHODS: On the medial side of the cleft, the short philtrum is lengthened and the upwardly displaced Cupid's bow maintained in a lowered position by a rectangular flap transplanted from the lateral surface of the columella. No lip flap has been formed on the lateral side of the cleft. RESULTS: In a period of three years (1996-1998), 23 cases of unilateral cleft lip, 9 cases of bilateral cleft lip and 14 cases of secondary deformities of cleft lip have been repaired with this new method. All of the repaired lips have normal looking philtrum column (limited by an uninterrupted straight-line scar), philtrum dimple and Cupid's bow. CONCLUSION: The lateral columellar flap method is a more rational surgical procedure for cleft lip repair in that it repairs tissue defect by transplanting tissue from the unexposed area. The appearance of the repaired lip is better than that of the commonly used methods. PMID- 11501090 TI - [Tissue response in circum-maxillary sutures after transverse palated suture expansion]. AB - OBJECTIVE: This study was carried out to investigate the histological reactions of midfacial sutures induced by transverse palatal suture expansion. METHODS: Nine mongrel dogs of 8-week old were used in the study. Six of them were used as experimental group as described in a previous paper. The additional three served as controls. In the experimental dogs, a NiTi-shape memory alloy (NiTi-SMA) arch wire was used to expand the transverse palatal suture. Tetracycline was injected intramuscularly (50 mg/kg) three days before operation and at the time of sacrifice. Tissue reactions of premaxillo-maxillary suture, transverse palatal suture, zygomatico-maxillary suture, zygomatico-temporal suture and fronto maxillary suture were observed in undecalcified tetracycline labeled sections and decalcified histological sections. RESULTS: The sutures in normal young dogs showed active bone formation. With the increase of age, the cellular components of the suture decreased. A great amount of new bone apposition was found in the bony front of expanded transverse palatal suture of the experimental dogs. Degenerative changes were found in premaxillo-maxillary suture at the expanded side while the contralateral showed active osteogenesis. Other sutures manifested both bone apposition and deposition due to the irregular interdigitations. CONCLUSION: After expansion, a great amount of new bone formed in the transverse palatal suture and thus the hard palate was lengthened. All the circum-maxillary sutures reacted in different patterns. PMID- 11501091 TI - [A new approach for secondary deformities of unilateral cleft lip nose]. AB - OBJECTIVE: The aim of this work was to correct the secondary deformities of unilateral cleft lip nose. METHODS: On the basis of patho-anatomical study and the comprehension on the cleft lip nose, a new approach to correct the secondary deformities of unilateral cleft lip nose was designed and used. During operation, an intranasal and marginal combining incision was made to explore the alar cartilages and to from a mucosal-cartilage flap in the nasal vestibule. After the deformity of septal cartilage and the abnormalities of the alar base on the cleft side were repaired, the total alar cartilage was repositioned and rotated with suspension and V-Y advancement to correct the nasal deformities. RESULTS: Since 1993, a total of 92 cases were treated by the above procedure and satisfactory results were obtained. CONCLUSION: The clinical applications showed that there were several advantages for this method, and good results can be gained by repositioning the total alar cartilage on the cleft side. PMID- 11501092 TI - [Intraoral one-stage curved osteotomy for prominent mandibular angle]. AB - OBJECTIVE: This paper presents a new method of osteotomy for the prominent mandibular angle (PMA). METHODS: Two modifications have been made to the conventional procedures. The first is the one-stage curved osteotomy of the mandibular angle. The second is non-resection of the masseter muscle. The surgical procedure has been performed through an intraoral approach with a direct field of vision. RESULTS: From December 1996 to December 1997, a total of thirty eight patients suffering from PMA have been operated on by this method with satisfactory results. The follow-up is 3 to 12 months. CONCLUSION: This method is a very simple and ideal procedure with much less complications for surgical treatment of PMA. PMID- 11501093 TI - [Repair of the skin defect on the remote part of the lower limb with the distally based sural nerve nutritional artery island flap]. AB - OBJECTIVE: The study was to verify the existence of the nutritional artery accompanying the sural nerve and to design a distally based island skin flap nourished by this artery for repairing skin defects on the remote part of the lower limb. METHODS: Anatomical study on 15 cadaver lower limbs and clinical applications in 7 cases were performed. RESULTS: The sural nerve nutritional artery is plexiform, forming anastomoses with the vascular network in the superficial and deep fascia. All 7 skin flaps depending on this arterial supply survived in clinical practice. CONCLUSION: This flap has high success rate. The operation is less traumatic and easy to perform. For the repair of the lower limb, it is a good option. PMID- 11501095 TI - [Re-assessment of the inframammary crease ligament]. AB - OBJECTIVE: An anatomical study was carried out to examine the existence of the inframammary crease ligament. METHODS: 18 breasts of preserved female cadavers were dissected and dyed for the fascia. RESULTS: The so-called inframammary crease ligament described by other authors does not exist. CONCLUSION: The reinforced Cooper's ligament supports the inframammary fold in position. PMID- 11501094 TI - [Classification and surgical correction of hump nose]. AB - OBJECTIVE: To establish a new classification of hump nose and to develop operative techniques based on its characteristics in Chinese patients. METHODS: This paper presents and summarises 68 cases of hump nose. According to the anatomic structure of the hump nose, we classify it into four types: small hump type I and II, moderate hump and large hump. Based on the classification of hump nose, two operative methods were developed-osteotomy and non-osteotomy. RESULTS: From 1990 to 1997, 68 cases of hump nose were operated on. Postoperative follow up for 1-18 months showed satisfactory results. CONCLUSION: By means of this classification and the operative techniques, good result can be obtained for the correction of the majority of hump noses in the Chinese. PMID- 11501096 TI - [Acute ischemic preconditioning protects against skeletal muscle infarction in the pig]. AB - OBJECTIVE: The aim of this study was to investigate metabolism of ischemic muscle and the efficacy of acute ischemic preconditioning for protection of skeletal muscles against infarction. METHODS: The efficacy of preconditioning was tested by subjecting pig latissimus dorsi muscle to 3 cycles of ischemia reperfusion, each for 10 min, before 4 h of global ischemia. Infarction was assessed at 48 h reperfusion using nitroblue tetrazolium dye. Muscle biopsies were taken from the latissimus dorsi before ischemia, at the end of 2 and 4 h of ischemia and 1.5 h of reperfusion. RESULTS: Preconditioning reduced the total infarct size by 44% in the latissimus dorsi. The muscle contents of ATP were maintained higher and the lactate lower (P < 0.05) in the preconditioned than in the non-preconditioned muscle at the end of 2 h, 4 h of ischemia and 1.5 h of reperfusion. CONCLUSION: Preconditioning of pig skeletal muscle is associated with a lower energy metabolism during sustained ischemia. At the present time, it is not known if this energy sparing effect is a major mechanism of ischemia preconditioning against infarction in the skeletal muscle. PMID- 11501097 TI - [Effects of heat-shock pretreatment on the survival of island flaps in rats]. AB - OBJECTIVE: This study was to develop a new method that can lessen ischemia reperfusion injury and improve the survival of the island flap. METHOD: A right lower abdominal island flap was created in the SD rat according to the protocol of Harashina. Animals were divided into 2 groups (heat-shock pretreatment and control groups). The island flap viability and electronic microscopical appearance of flap tissue were evaluated following ischemia reperfusion injury. In order to explore the mechanism of this phenomenon, we examined the contents of HSP70 and measured the levels of SOD and MDA of the flap at various times. RESULTS: Compared with the control group, SOD activities of the flap were higher, and MDA content was lower. Survival rate of the island flap with heat-shock pretreatment was significantly increased (P < 0.01). CONCLUSION: The heat-shock pretreatment can lessen ischemia reperfusion injury and improve the survival of the island flap. It is suggested that the mechanism be related to following aspects: 1. The heat-shock pretreatment maintains stability of cell's function and structure. 2. It may increase the ability to catabolize free radicals by antioxidative enzymes. PMID- 11501098 TI - [High voltage transmission electron microscope and X-ray double-crystal diffractometer observation of pure titanium implant-bone interface]. AB - OBJECTIVE: To examine the integration of pure titanium implant and bone tissue. METHODS: Using low temperature dualion mill technique, the interface of titanium implant and bone was examined with high voltage transmission electron microscope and X-ray double crystal diffractometer. RESULTS: It was shown that calium globules of various electron density or sheets of calcified osseous tissue blended with titanium at the interface. The bone had grown in two directions. CONCLUSION: The integration of pure titanium implant and bone tissue is good, leaving no gap at their interface. PMID- 11501099 TI - [Effects of microwave baking on the immunological cells in primary lymphedema patients]. AB - OBJECTIVE: To elucidate the effects of microwave on the immunological cells in primary lymphedema. METHODS: The immunological cells including lymphocytes in the affected limb skin and peripheral blood of 10 patients with primary lower limb lymphedema were analysed using ABC and APAAP immunohistochemical methods before and after microwave baking and bandaging treatment. RESULTS: It is demonstrated that in the peripheral blood of the patients there was an increase of CD8+ T lymphocytes as well as a decrease of CD4+ T lymphocytes and the ratio of CD4/CD8. It was found that there was significant perivascular infiltration of mononuclear cells (most were monocytes and macrophages) in the skin of the affected limb. CONCLUSION: Microwave modulates the systemic immunological imbalance by its heating and complex biological effects on primary lymphedema patients through reversing the ratio of CD4/CD8 to normal level by increasing CD4+ T lymphocytes and decreasing CD8+ T lymphocytes. It can also decrease the perivascular T lymphocyte infiltration of the affected dermis and enhance the phagocytic capabilities by promoting the proteolytic activities of macrophages, finally resulting in edema resolution. PMID- 11501100 TI - [Role of endothelin and nitric oxide in early gut ischemia]. AB - OBJECTIVE: Inadequate perfusion in splanchnic organs and especially in the gut during acute burn period has been reported in many conventionally "successfully" resuscitated patients, but the mechanisms still remain unclear and its early preventive measures need to be further studied. The aim of this study is to evaluate the role of endothelin and nitric oxide in gut ischemia. METHODS: Eighteen male pigs were randomly assigned to one of the three groups: group C, a sham burn group that was subjected to all surgical procedures except burn; group B, sustained 30% TBSA cutaneous thermal burn; Group N, NO donor (C87-3754) was given intravenously (0.0125 mg.kg-1.min-1) at the beginning of resuscitation. RESULTS: In group B, PVF decreased rapidly after burn, and did not recover in the observation period (72 h), ET levels in portal blood and intestinal tissue elevated contrary to the changes in NO. In group N, PVF was higher than in group B. CONCLUSION: 1. Changes in ET and NO may influence the protal blood flow. 2. NO donor was proved to be beneficial in improving GI tissue perfusion by releasing NO. PMID- 11501101 TI - [The effects of zinc on liver metallothionein in scalded rats]. AB - OBJECTIVE: To investigate the changes in liver metallothionein (MT) in scalded rats. METHODS: The scalded rats in deficient zinc condition were fed with diets containing zinc 4 micrograms/g, 10 micrograms/g, and 80 micrograms/g respectively. The changes in serum zinc, liver zinc and liver MT were observed. RESULTS: The serum zinc decreased, liver zinc and MT increased. The higher diet zinc levels were, The higher serum zinc, liver zinc and MT became. The correlation analysis showed that the changes between liver zinc level and liver MT was positive relationship(r = 0.85, P < 0.01). CONCLUSION: Serum zinc decrease, liver zinc increase after burn. This is connect with increment of MT synthesis in liver, zinc is inducer to MT. It can improve synthesis of liver MT. PMID- 11501102 TI - [The changes in endogenous growth factors and their relation with wound healing]. AB - OBJECTIVE: To explore the effects of endogenous growth factors on the formation of scars and ulcers after trauma or burn. METHODS: The amounts of endogenous epidermal growth factor (EGF), tumor necrosis factor(TNF) and nitric oxide (NO) in scar and the developing granulation tissues after trauma or burn were determined and their relation with the wound healing were studied. RESULTS: The results showed that EGF, TNF and NO could be found in all scars and the granulation tissues, but their concentration varied with different tissues, sex and age of patients. In male patients or youngsters, the amounts of EGF, TNF and NO in scars were much greater than those in granulation tissues. CONCLUSIONS: The results indicated that the growth factors are necessary for wound healing, but insufficiency or overproduction of the endogenous growth factors may lead to the development of chronic ulcers or scar formation after trauma or burn. PMID- 11501103 TI - [The effect of the molecular triggers of hemostatic activity on thrombosis in early stage of electrical injuries]. AB - OBJECTIVE: To evaluate the mechanisms of thrombosis in the early stage of electrical injury. METHODS: Endothelin-1(ET-1), prostacylin(PGI2), platelet alpha granule membrane protein (Gmp-140), thromboxane (TXB2), and plasminogen(PLG-A) were measured in 26 patients with electrical injury. RESULTS: It was found that GMP-140 and ET-1 increased significantly(P < 0.01), PLG-A showed a significant change 2 weeks after the injury, while there was an imbalance between TXB2 and PGI2. CONCLUSION: It is believed that change in one or several mediators mentioned above may trigger thrombosis after electrical injury. PMID- 11501104 TI - [The changes in arteriography in extremities sustaining high tension electric injuries]. AB - OBJECTIVE: Because it is difficult to estimate the extent and degree of damage to the blood vessels of extremities sustaining high tension electrical injuries, arteriography was carried out to evaluate the vascular changes. METHODS: Selective arteriography was carried out 29 times for 26 extremities in 17 patients suffering from > 380 V electrical injuries. RESULTS: The arteriography showed that the injured arteries were narrowed and occluded, and vascular branches decreased or disappeared. Recanalization of the partially occluded arteries or collateral circulation formation was found after decompression. CONCLUSION: Arteriography is valuable to evaluate the vascular changes in high tension electrical injuries of extremities. PMID- 11501106 TI - [Endothelial cell damage induced in vitro by subeschar tissue fluid]. AB - OBJECTIVE: To assess the effect of subeschar tissue fluid (STF) on the function and structure of endothelial cell in vitro. METHODS: Human umbilical vein endothelial cells (HUVEC) were incubated with STF obtained from patients in the early postburn stage. Then the morphological changes in EC were observed, and activity of EC and contents of LDH, 6-keto-PGF1 alpha in media were determined. RESULTS: The activity of LDH and the content of 6-keto-PGF1 alpha were increased gradually after HUVEC incubated with STF, and were higher than that measured after HUVEC incubated with healthy volunteer's sera. Furthermore, activity of HUVEC decreased and they became deformed, showing shrinkage of the cell body with enlargement of intercellular space. In addition, small vacuoles appeared in cytoplasm with karyopyoknosis after 24 h. CONCLUSION: STF can damage endothelial cell and might play an important role in the pathophysiological process of burn injury. PMID- 11501105 TI - [Clinical studies on the effects of burn trauma on pituitary-testis axis]. AB - OBJECTIVE: To investigate the effects of burn trauma on pituitary-testis axis. METHODS: The changes in serum estradiol(E2), testosterone(T), luteinizing hormone(LH), and follicle stimulating hormone(FSH) levels were dynamically investigated by RIA in 30 days after thermal injury in 28 burnt men. RESULTS: The serum FSH, LH, and T levels significantly decreased after injury, and there was some increase from PBD(postburn day) 14 to 30. In patients in group II and III (BSA > 30% TBSA), not all of their FSH, LH, and T levels reached the normal control on the PBD 30. Serum E2 levels were elevated after injury, then returned gradually to normal. In three patients who died, their mean T levels were significantly lower than patients who survived, but mean E2 levels were significantly higher than that of survivors. CONCLUSION: The activity of pituitary might be depressed after burn trauma. The imbalance between E2 and T might play a role in the processes of immuno-depression after injury. The changed extent of E2 and T levels was correlated to burn size. They could be sensitive indicators of burn stress. PMID- 11501107 TI - [Progress in the study of immunological activity of silicone gel]. PMID- 11501108 TI - [Clinical uses of growth hormone in burns]. PMID- 11501109 TI - [The study of wound repair]. PMID- 11501110 TI - [Study on the expression of calreticulin in hypertrophic scar-derived fibroblasts]. AB - OBJECTIVE: To investigate the expression level and intracellular distribution of calreticulin(CRT) in hypertrophic scar-derived fibroblasts(HSF) and normal skin derived fibroblasts(NSF). METHODS: Using anti-CRT antibody, CRT was detected in HSF and NSF by immunocytochemistry and ELISA technique. RESULTS: CRT was present in the cytoplasm and nuclei in proliferating HSF or NSF in culture, but HSF possessed significantly higher expression level of CRT than NSF(P < 0.01). When these cells (HSF and NSF) were fused, neither intracellular nor intranuclear staining for CRT was observed. CONCLUSION: In HSF and NSF, the distribution of CRT is widespread. Because of its abundant expression in HSF, we postulate that CRT as a multifunctional protein plays an important role in Ca2+ sequestering, integrin-mediated signalling and cell adhesion. PMID- 11501111 TI - [Effects of PCMV4-hTGF beta 1 as nucleic acid vaccine on II* burn wound healing and postburn scarring in rats]. AB - OBJECTIVE: To investigate the gene expression of PCMV4-hTGF beta 1 as nucleic acid vaccine regulation and the effects on burn wound healing and scarring in rats and study the feasibility of gene therapy in burns. METHODS: Sixty wistar rats were divided into three groups: 1. burn + nucleic acid vaccine; 2. nucleic acid vaccine only(control); 3. burn only(control). ELISA was used to determine the dynamic changes in anti-TGF beta 1 neutralizing antibody in serum; determination of the quantities of DNA retained in muscles by Southern blot and in situ hybridization; observation of the ratio in collagen I/III during the wound healing by special stain method. RESULTS: The level of anti-TGF beta 1 antibody in serum reached the peak at the third week after naked DNA was injected, and a slight drop in the 4th week. TGF beta 1 plasmid DNA could be detected 5 minute after injection, and lasted 3 hours. In situ hybridization showed a positive staining in muscle fibers 5 days after injection. During the day 0-day 9, the wound healing speed in vaccine group was faster than control, and after day 10, no significant difference was found between groups, but the ratio of collagen I/III was reduced remarkably in vaccine group. CONCLUSION: Injection of PCMV4-hTGF beta 1 in rats can really cause general immune response reaction. It was showed that PCMV4-hTGF beta 1 was similar to TGF beta 1, and it had the effect of stimulating of epidermic cells to accelerate wound healing at early stage, and at later stage, it was similar to anti-TGF beta 1 neutralizing antibody, having the effect of inhibiting hyperplasia of scar. So it is confirmed that PCMV4-hTGF beta 1 as nucleic acid vaccine has the effect of promoting healing of burn wound and controlling the formation of scar. PMID- 11501112 TI - [The effect of intradermal injection of collagenase in implanted hypertrophic scar in nude mice]. AB - OBJECTIVE: Effects of collagen degradation in hypertrophic scar by intradermal injection of collagenase were observed. METHODS: Hypertrophic scar was implanted into nude mice and collagenase was injected locally. Light and electron microscopic observations were carried out. RESULTS: Collagen degradation in hypertrophic scar by intradermal injection of collagenase was observed. The implanted scars became soft, thin and small. CONCLUSION: These results suggested that local injection of collagenase may be a useful therapeutic tool for hypertrophic scar. PMID- 11501113 TI - [A study on guided tissue regeneration and tissue engineering-transplantation of collagen membrane seeded with cultured hyaline chondrocytes]. AB - OBJECTIVE: Lack of transplant material is a main problem in the repair and reconstruction of tissue or organ defects. To seek the succedaneum of transplants, we studied and used tissue-engineered growth hyaline cartilage. METHODS: The articular hyaline cartilage obtained from newborn calf within 6 hours of sacrifice was enzymatically dissolved. The chondrocytes were then seeded onto the medical collagen membrane of guided tissue regeneration in a 24 well plate in RPMI1640 medium. One week later, the chondrocyte medical collagen membrane of guided tissue regeneration complex was implanted subcutaneously into the back of nude mice. Eight weeks after implantation, the animals were sacrificed. RESULTS: The present study seems to demonstrate that the cartilage like tissue was strong enough to be transferred after being implanted for 8 weeks. The cartilage-like tissue was proved to be tissue-engineered hyaline cartilage by HE stain. The chondrocytes could secrete chondroitin sulfate as proved by Lev-Spicer stain. CONCLUSION: Tissue engineering technique was used to make tissue-engineered cartilage in vitro. It will provide a new transplant material for the repair of tissue or organ defects. PMID- 11501114 TI - [Culture of chondrocytes using allogenous acellular cartilaginous matrix]. AB - OBJECTIVE: To observe the effect of chondrocyte culture with natural ECM made from allogenous acellular cartilage. METHODS: The allogenous acellular cartilaginous matrix was used as a scaffold in chondrocyte culture in vitro, comparing with the acellular perichondrium. The growth and regeneration of the chondrocytes in vitro were observed. RESULTS: The chondrocytes seeded on the acellular cartilaginous matrix grew well while the chondrocytes on the acellular perichondrium matrix grew poorly. CONCLUSION: The surface condition of the scaffold has a notable influence on the growth of the cells. The allogenous acellular cartilaginous matrix may provide a suitable condition for chondrocytes, and it has the possibility to be developed as a natural scaffold for tissue engineering. PMID- 11501115 TI - [Repair of rabbit auricular defects using chondrocyte-scaffolding constructs]. AB - OBJECTIVE: To evaluate if the tissue-engineered cartilage can repair the auricular defects in rabbits that have inherent immunity. METHODS: Rabbit auricular defects were repaired using chondrocyte-PGA constructs cultured in vitro. The gross and histological comparisons were made between the experimental and the control groups. RESULTS: The rabbit auricular defects were repaired with chondrocyte-PGA constructs, leaving fibroid tissue in the interface between the tissue-engineered cartilage and the normal cartilage. CONCLUSION: The auricular defects can be repaired by chondrocyte-PGA in vitro cultured constructs in animals like rabbits that have acquired immunity. The interface healing problems need further study. PMID- 11501116 TI - [Clinical use of the prefabricated expanded island skin flap]. AB - OBJECTIVE: This is to introduce the applications of the prefabricated expanded island skin flap based on a fasciovascular pedicle. METHODS: The superficial temporal vessel was transferred to the cervical region and an expander was buried under it. When the expander was fully expanded, an island skin flap was elevated and transferred based on the fasciovascular pedicle. Eight patients were treated with this method since 1996. RESULTS: All the flaps survived without necrosis. CONCLUSION: The island skin flap prefabricated in this manner has rich blood supply and can be transferred safely. It is useful in plastic and reconstructive surgery, especially for the severe burned patients whose normal skin is insufficient. PMID- 11501117 TI - [Coverage of the lower leg defects using the reversed island skin flap based on the nutrient vessels of the saphenous nerve]. AB - OBJECTIVE: To report the application of the distally based neurocutaneous saphenous island flap for the coverage of soft tissue defects in the lower limb. METHODS: According to the size and site of the defect and the rotation point, the flap was designed, and it was based on the arterial axis associated with the saphenous nerve and the greater saphenous vein. RESULTS: Six neurocutaneous saphenous island flaps were used and all survived. Flap dimensions were as large as 6 cm x 8 cm. The results were satisfactory after 6 to 20 months' follow-up. CONCLUSION: The vascularity of the saphenous nerve is closely connected with the vascularity of the skin. The flap supplied by the accompanying vessels of the cutaneous nerve can be utilized with good results. PMID- 11501118 TI - [Clinical application of 96 hours' rapid skin tissue expanding technique]. AB - OBJECTIVE: To reduce expansion course, diminish complications and improve expanding effect in skin tissue expander treatment. METHODS: We have chosen 38 cases of skin tissue defects of small and moderate areas, on which we have carried out 96 hours' rapid skin tissue expanding technique by altering the time and the amount of saline injected. RESULTS: All skin tissue defects have been successfully repaired. Half a year later, the incisions were inconspicuous. CONCLUSION: 96 hours' rapid skin tissue expanding technique has the following advantages, it is simple and safe, economic and convenient, rapid and reliable. This technique tends to be mature. What is more, it is easy to be popularized. We regard it as an ideal rapid expansion technique. PMID- 11501119 TI - [Suture expansion osteogenesis for closing the posterior hard palate: an experimental study in dogs]. AB - OBJECTIVE: This study investigated the possibility of a new approach, e.g. suture expansion osteogenesis for closing the posterior hard palate. By this technique we might hope to replace missing tissue in the clefts by tissue regeneration and achieve bony repair in the posterior hard palate and circumvent the problems of speech and facial growth seen in traditional cleft surgery. METHODS: Nine 8-month mongrel dogs were divided into three groups: the control(n = 1), sham control(n = 2) and the experimental group(n = 6). In six experimental dogs, an 8 mm wide cleft was made surgically. Suture expansion devices made of NiTi-shape memory alloy with force levels of 200 g, 360 g and 480 g were secured in the palatine bones to expand the lateral sutures of the palatine bones. Clinical and histological examinations were performed to observe the changes. RESULTS: The clefts in the experimental group were closed completely sooner or later within a period from 5 to 14 days. While the clefts in the sham group became a little larger than their original size. There was significant new bone formation at the edge of the expanded suture. CONCLUSION: The clefts in young dogs could be closed by the technique of suture expansion. The NiTi-SMA expander used in the study was consistently effective and all the force levels could induce a great amount of new bone formation with the trabeculae being thinner and longer by heavy forces. PMID- 11501120 TI - [Comparative study on early revascularization of membranous and endochondral onlay bone grafts in the rat]. AB - OBJECTIVE: To investigate the disparity in early revascularization of the membranous and endochondral bone grafts. METHODS: Twenty Wistar rats were randomly divided into two groups of 10 each. Full-thickness cranial (membranous) and split-thickness iliac crest (endochondral) onlay bone grafts were placed subperiosteally onto the snout of the animal without bony fixation. At 7 and 14 days after grafting the bone grafts together with the recipient sites were harvested. Revascularization was studied utilizing opacifying injection and computerized image analysis technique. RESULTS: The extent of vascularization at 7 days was 11.88% and 22.33% in membranous and endochondral bone grafts, respectively, and they developed to be 15.93% and 34.93% at 14 days, respectively. CONCLUSION: The results indicated that endochondral bone grafts were more rapidly revascularized than membranous bone grafts at 7 and 14 days (P < 0.05). Differences in graft architecture are theorized to account for the relationship between the vascularization and volume maintenance in the bones of different embryonic origin. PMID- 11501121 TI - [Human preadipocyte culture and the establishment of hyperplasia and hypertrophy model]. AB - OBJECTIVE: To better understand hyperplasia and hypertrophy properties of human adipose tissue. METHODS: Using an "adhesion-ceiling" culture strategy, fibroblast like cells from adult pure adipose granules were successfully cultured. RESULTS: The cells were highly homogeneous, highly proliferative and with a high differentiation rate. Their dynamic morphological changes, growth curve, glycerophosphate dehydrogenase pattern, oil red O staining, and reaction to insulin and dexamethasone all verified their preadipocyte identity. Under controlled conditions, the preadipocytes replayed their in vivo hyperplasia and hypertrophy process. CONCLUSION: In mature human adipose tissue there exist functionally active fractions that can be modified. This study laid the basis for further probing into obesity and fat autotransplantation. PMID- 11501123 TI - [Comparison between copper needle insertion and stainless steel needle insertion into rabbit ear central veins]. AB - OBJECTIVE: The clinical application of copper needle insertion has been proceeded satisfactorily for peripheral vascular disorders. This work was to study the mechanism of the copper needle insertion on a pathological basis. METHODS: Sixty rabbits were used in the experiments. The copper needle or the stainless steel needle was inserted into the ear central vein of the animal. Then the specimens of the central vein including its surrounding tissue were taken for light and electron microscopic examinations. RESULTS: At 7 days after copper needle insertion, acute inflammation reaction of the venous wall and vascular thrombosis were revealed in the specimens. At 14 days, giant cells granuloma and destruction of the venous wall were seen. After 28 days to 2 months, granuloma and the residual destroyed venous wall were gradually absorbed and fibrosis resulted. The vascular inflammation and thrombosis caused by stainless steel needle insertion was slight and slow. CONCLUSION: Copper needle treatment for vascular disorders is superior to the stainless steel needle. PMID- 11501122 TI - [Experimental study and preliminary clinical application of bacterial collagenase in catabolizing scars]. AB - OBJECTIVE: To investigate the mechanism of collagenase in degradation of hypertrophic scars and observe the clinical effects of this therapy. METHODS: Both animal experiment and clinical trial were conducted. RESULTS: The injection of collagenase into the hypertrophic scar tissues in nude mice resulted in 86% volume reduction after two injections within two weeks, while the control tissues reduced only by 35% during the same time. Clinically, 13 cases were treated with intralesional injection of bacterial collagenase. The volume reduction rate was 46.92% in average within two weeks. The volume reduction in 4 of 13 cases was above 50%. Three to sixteen months' follow-up of five patients revealed that only one case of keloid recurred after treatment in the second month. Examinations of histological sections and TEM showed the dissolved collagen fiber clearly. CONCLUSION: Collagenase can degrade collagen fiber directly. The prompt significant effects and the mild complications make it a prospective therapy. PMID- 11501124 TI - [Study of surgical depilation in pig and human body]. AB - OBJECTIVE: The study is to show how much dermis and the follicles within it should be removed in order to prevent hair regeneration. METHODS: The experiment used Wuzhishan mini pigs. Different thickness of the pig's dermis and the follicles within it were removed to observe hair regeneration. According to the result, the dermis and the follicles were removed from the scalps of 62 patients from Jan. 1995 to Dec. 1996. Subsequent hair regeneration was observed. RESULTS: Using surgical depilation to remove 1/4-2/5 thickness of the scalp dermis and the follicles within it stopped most of the hair regeneration. The rate of hair regeneration was 10-30%. Satisfactory clinical effects were achieved. CONCLUSION: Surgical depilation is a simple, effective, cheap way for hair removing. PMID- 11501125 TI - [Analysis of factors influencing healing time of burn wounds]. AB - OBJECTIVE: To analyze the factors influencing healing time, length of hospital stay and hospital costs in burn patients. METHODS: Seven hundred and ninety-one burn patients with principal component analysis were studied retrospectively. RESULTS: There were some factors that influence healing time, including total area of burn, the area of full-thickness burn, the area of partial-thickness burn, inhalation injury, sepsis, shock, visceral complication, combined injuries, the size of surgical excision, the time of surgical excision, whether or not excision of burn wound was done in shock stage, and use of antibiotic, etc. Further analysis indicated that the patients undergo escharectomy during shock stage had better recovery with lower incidence of visceral complications and sepsis, especially in those with major burn over 30 percent TBSA. CONCLUSION: To reduce healing time of burn patients, there are many effective methods such as the prevention and treatment of shock, sepsis, visceral complication, and appropriate use of antibiotics. The most important factor to reduce healing time of burn patients is escharectomy as early as possible for major burns. PMID- 11501126 TI - [The study of growth hormone on wound healing rate in adult burns]. AB - OBJECTIVE: To determine the effects of rhGH on wound healing in adult burn patients. METHODS: 42 patients with burn wounds covering over 83% of total body surface area were enrolled in this placebo controlled prospective study. They were comparable in nutrient intake, TBSA, and full thickness burn area. For rhGH group patients rhGH subcutaneously was given in the dose of 0.2 U.kg-1 at 8 a.m. each morning for 14 days. The control group patients were given normal saline as placebo. Protein catabolism, immunocompetence, healing time of burn wounds and donor sites were assayed. RESULTS: 1. The patients in rhGH group had higher serum levels in albumin, transferrin, prealbumin and CD4 T-cells (P < 0.01). 2. The healing time of autologous skin grafting and donor site and the length of hospital stay were significantly shorter in rhGH group patients compared with control group. CONCLUSION: rhGH could enhance the wound healing rate, improve protein anabolism and reduce the length of hospital stay in severe burn patients. PMID- 11501127 TI - [Fiberoptic bronchoscopy in diagnosis and treatment of inhalation injury]. AB - OBJECTIVE: To explore the morphological standard of diagnosing inhalation injury, the feasibility of early diagnosis and the topical therapy of inhalation injury through fiberoptic bronchoscopy. METHODS: In one hundred and forty four patients with inhalation injury that were admitted in 10 years were examined with bronchoscopy, and the results were analysed. RESULTS: Diagnostic criteria and classification were obtained with the aid of the bronchoscopy. It was found feasible to make early diagnosis within 8 h and also to exercise treatments in the form of suction, irrigation, etc. CONCLUSION: Bronchoscopy plays an important role in the early diagnosis and treatment of inhalation injury, and it should be used more extensively in patients suspected to have inhalation injury. PMID- 11501128 TI - [Effects of hyperbaric oxygen therapy on the changes in serum sIL-2R and Fn in severe burn patients]. AB - OBJECTIVE: Many reports showed that level of soluble interleukin 2 receptor (sIL 2R) in sera was obviously increased in burn patients, while plasma fibronectin (Fn) was markedly decreased, and they had close relation with the occurrence and development of burn infection. The purpose of this study was to verify the significance of hyperbaric oxygen therapy (HBOT) in the prevention and treatment of burn infections by observing its effect on the levels of serum sIL-2R and Fn in severe burn patients. METHODS: Forty two burn patients(with more than 30% body surface area or three degree more than 10%) were selected and divided into two groups at random: HBOT group(25 cases) and non-HBOT group(17 cases); forty healthy blood donors were selected as normal control. The venous blood samples were drawn from 42 patients at 1/3, 1, 2, 3, 5, 7, 10, 14, 17, 21, 28 and 35 days postburn, the levels of serum sIL-2R and Fn were measured by ELISA and rocket electrophoresis techniques respectively, meanwhile, the incidences of sepsis of two groups were compared. RESULTS: 1. In non-HBOT group: the levels of serum sIL 2R were remarkably increased, while serum Fn were markedly decreased at all periods postburn (compared with normal control: P < 0.05 or P < 0.01); 2. In HBOT group: the levels of serum sIL-2R were not obviously increased at all periods except 21 days postburn, and Fn were not decreased at all periods except 8 h postburn (compared with normal control: P > 0.05). Compared with non-HBOT group, however, the levels of serum sIL-2R were profoundly reduced, while Fn was significantly enhanced at all periods postburn(P < 0.05 or P < 0.01), and the incidence of sepsis was obviously lowered (P < 0.05). CONCLUSION: HBOT can significantly reduce the level of serum sIL-2R but enhance Fn in severe burn patients, so it may have a positive effect on the prevention and treatment of burn infections. PMID- 11501130 TI - [Gene therapy technique and its uses in tissue repair]. PMID- 11501129 TI - [Early cardiac myocyte membrane changes in severely burned rats: a spin labding ESR study]. AB - OBJECTIVE: To demonstrate the mechanism of early cardiac myocyte membrane damage at molecular level in severely burned rats. METHODS: SD rats were randomly divided into sham-burn and burn groups. A 30% TBSA burn was created in burn group. Hearts were obtained at different time postburn. Cardiac myocytes isolated by collagenase using Langendoff perfusion apparatus were spin labeled with 5 doxylstearic acid (5NS), 16-doxylstearic acid (16NS) and 3-maleidoproxyl (3MP). The order parameters (S value), hs/hw ratio and rotational correlation time (tau c) were measured according to the spin labeling-ESR spectra. RESULTS: The S value of the polar head-group region of membrane lipid, as reported by 5NS, did not changed significantly, while that of the lipid in the hydrophobic midzone, as reported by 16NS, increased significantly. The hs/hw ratio of 2 and 6 hours postburn groups increased significantly postburn compared with that of the sham burn group, and so did the rotational correlation time in 1 to 12 h postburn groups. CONCLUSION: There were structural and functional changes in cardiac myocyte membrane after burn injury. They were characterized by decreased fluidity in the deeper part of membrane lipid, configurational changes and motion restriction in membrane proteins. They could affect the normal performance of cardiac function. PMID- 11501131 TI - [Brief introduction to tissue engineering]. PMID- 11501132 TI - [Changes in cardiac renin-angiotensin system after severe burn injury in rats]. AB - OBJECTIVE: To observe changes in cardiac renin-angiotensin and its effect on sarcoplasmic reticulum(SR) calcium transport function, and investigate the mechanism underlying cardiac dysfunction in early postburn stage. METHODS: Rats were divided into three groups: control group (n = 8, without burns), burn group (n = 40, 30% TBSA full-thickness burns), and treatment group (n = 40, gavage Lisinopril for 3 days before 30% TBSA burns). The left ventricular(LV) contractile function was measured at 3, 8, 24 hours postburn. Angiotensin converting enzyme(ACE) activities, the contents of angiotensin II (AII) and calcium in myocardium were also measured. In addition, Ca(2+)-ATPase activities and calcium transport function in SR were determined. RESULTS: The maximal rates of LV pressure rise (+/- dp/dtmax) and fall (-dp/dtmax) in burn group were significantly lower than that in control group. The ACE activities, AII levels, as well as calcium contents in myocardium increased markedly in burn group. Ca(2+)-ATPase activities and Ca2+ uptake function in SR decreased significantly in burn group. After treatment with ACE inhibitor-Lisinopril, the ACE activities, AII levels and calcium contents were significantly decreased compared to the burn group. Meanwhile, Ca(2+)-ATPase activities, calcium uptake function and LV contractile function decreased markedly. CONCLUSION: The cardiac renin angiotensin system is activated rapidly after severe burns and inhibits the calcium transport function which may play an important role in cardiac contractile dysfunction following burns. PMID- 11501133 TI - [Effect of different immunomodulators on macrophage function in rats with scald injury]. AB - OBJECTIVE: To explore effect of different immunomodulators on macrophage (M phi) functions in rats with scald injury. METHODS: The M phi functions in various groups of rats with scald on the 5th and 10th day after scald were respectively determined with methods of McAb APAAP, agar bacteriolytic plate and MTT colorimetry. RESULTS: The results showed that: 1. after scald, presenting rate of Ia antigen, ability of antigen presentation, phagocytic power to candida albicans and lysozyme ability of M phi were significantly decreased, the capacity of M phi secreting TNF was markedly increased compared with those of the normal group, and the differences were significant (P < 0.01); 2. after the therapy with immunodulators, the above functions of M phi in rats with scald improved markedly. As compared with those of the control group, differences were significant (P < 0.01). CONCLUSION: The specific immune RNA, which is administered by intraperitoneal injection early after scald, can markedly improve immune functions of rats with scald. PMID- 11501135 TI - [The protective effects of prompt escharectomy on myocardial injury following burn in rats]. AB - OBJECTIVE: To investigate the protective effects of prompt escharectomy on myocardial injury following severe burns. METHODS: Rats were inflicted with 30% TBSA III degree scald. They were randomly divided into 3 groups: control group (C, n = 10), non-escharectomy group (NE, n = 50), escharectomy group (E, n = 50). In the E group, burn wounds were excised promptly after the injury. The levels of troponin T (TnT) and tumor necrosis factor (TNF) in plasma and TNF in myocardium were measured at 1, 3, 6, 12 and 24 hours postburn. RESULTS: TnT levels increased markedly at 3 hours post burn, plasma TNF levels increased markedly at 6 hours postburn, and TNF levels in myocardium also increased markedly at 12 hours postburn compared with the C group. Although the levels of TnT and TNF were a little lower in NE group than those in E group at 3 hours post burn, the levels of TnT 6 hours postburn and levels of TNF 12 hours postburn were significantly higher in NE group than those in E group. There were good positive correlations between TNF with TnT levels. CONCLUSIONS: TNF played important roles in the pathogenesis of myocardial injury following burns, its level was correlated with the severity of myocardial injury. TNF levels in E group was much lower that in NE group, suggesting that prompt escharectomy could reduce the production and release of inflammatory cytokines, and lessen the myocardial injury induced by uncontrolled inflammatory response. PMID- 11501134 TI - [Experimental study of cultured autologous keratinocyte sheet(Auto-CKS) and allodermis composite skin grafted onto open wound of rats]. AB - OBJECTIVE: Investigate the "take" and histological changes of grafted auto-CKS and allo-dermis composite skin. METHODS: Thirty Sprague-Dawley rats were divided into group auto-CKS and group allo-dermis composite skin. All wounds were dressed after grafting. Wound healing and "take" of grafts were determined for 90 days after grafting. Biopsies of wound tissue including grafts were collected at regular intervals for histological analysis to check the reconstruction of epidermis-dermis junction (EDJ) and fate of allo-dermis of the composite skin. RESULTS: In group auto-CKS, the "take" of grafts was pretty good, but the quality and histological structure of CKS was not good enough, the proliferated collagen fibers were disorderly arranged, and the CKS was too thin to withstand abrasion and tended to worn out and contracted severely. On the other hand, the "take" as well as the quality, histological structure, abrasion resistance and contraction of composite skin in the allo-dermis composite skin group were much better than that of group auto-CKS. Up to 90 days after grafting, the allo-dermis still existed without obvious signs of immuno-rejection. CONCLUSION: The CKS was not suitable for the wound of full thickness skin defect, while the composite skin composed of auto-CKS and treated allo-dermis unfolded the potential encouraging prospects. PMID- 11501136 TI - [Management of destructive electrical burns of wrist]. AB - OBJECTIVE: To reduce amputation rate of most severe pattern (Type III) of electrical burns of wrist. METHODS: Early wrist decompression followed by selective transfusion of dextran, PGE1, low molecule heparin to prevent thrombosis of injured radial and ulnar arteries, vascular bridging across the distal forearm and wrist to the hand soon after the appearance of circulatory failure to the hand. Free tissue transfer, especially the use of greater omentum and the innovated abdominal flaps, to repair the circumferential wounds of the wrists are recommended. RESULTS: A group of 102 limbs of 90 patients were treated in the last 10 years, among them the amputation rate of most severe pattern (Type III) of electrical burns of wrists was significantly reduced from 80% to 38.9% compared with 10 years before. CONCLUSION: A comprehensive treatment including the use of anticoagulation drugs, vascular reconstruction, sophisticated surgical repair of the wound are effective in reducing the amputation rate of electrical burns of wrists. PMID- 11501137 TI - [Hemodynamic monitoring in 52 serious burn patients in ten years]. AB - OBJECTIVE: To determine the benefit of hemodynamic monitoring for resuscitation of burn shock. METHODS: A group of 52 burned patients with mean burn area of (69.9 +/- 20.1)% TBSA burn injury (range 31%-100%), and mean III burn area of (60.4 +/- 13.6)% TBSA were included in this study. Swan-Ganz catheters were inserted to monitor hemodynamics including RAP, PAP, PAWP, HR, CO, CI, and SI, etc. after admission at 8, 16, 24, 36, 48, 72, and 96 hours postburn. RESULTS: It was reasonably safe to perform invasive monitoring during early resuscitation. With the guide of hemodynamic monitoring, evidence of global hypovolemia disappeared at 24 hours after burn injury with appropriate resuscitation therapy. CONCLUSION: Invasive hemodynamic monitoring may be necessary to optimize resuscitation of serious burn patients with reasonable safety. PMID- 11501139 TI - [Reconstruction of perforating defects of the maxillofacial region with double flaps]. AB - OBJECTIVE: To solve the problems in restoration of perforating (full thickness) defects in the maxillofacial region. METHODS: Double flaps have been used to reconstruct the defects. Forty-five patients with full thickness defects on the maxillofacial region were treated with 90 flaps, including 58 (64%) deltopectoral flaps, the platysma myocutaneous flap, pectoralis major myocutaneous flap, sternomastoid myocutaneous flap, forehead flap and latissimus myocutaneous flap. RESULTS: The operation results were satisfactory. Of the 90 flaps, 87 flaps survived completely or subtotally with a successful rate of 97%. Three flaps sustained total or large necrosis. CONCLUSION: A number of flaps can be used for repairing the full thickness defects of the maxillofacial region, especially the deltopectoral flap, the platysma myocutaneous flap and the pectoralis major myocutaneous flap. They can provide excellent external cover or internal lining. They are well vascularized, hairless and colour-matched. PMID- 11501138 TI - [Comparison of internal fixation with mini-titanium plate and wire osteosynthesis in midface fractures]. AB - OBJECTIVE: To compare clinical results of the internal fixation with wire and the internal fixation with mini-titanium plate in midface fractures. METHODS: By the coronal approach and supplementary incisions, forty patients with midface fractures were treated with wire osteosynthesis or rigid internal fixation (RIF) with mini-titanium plate. RESULTS: A good stability can not he obtained with wire osteosynthesis in the treatment of midface fractures. The supporting bony structure of midface can be reconstructed with mini-titanium plate RIF, which results in a sufficient three-dimensional stability of the fractured segments and an ideal facial configuration Mini-titanium plate RIF is more convenient and reliable than wire osteosynthesis in the difficult operation fields of the infraorbital margin and zygomaticomaxillary suture. CONCLUSION: Mini-titanium plate RIF is the best method for treatment of midface fractures. PMID- 11501140 TI - [A study of innervation of musculus temporalis and its clinical application]. AB - OBJECTIVE: To study the innervation of the musculus temporalis and improve the successful rate of the operation for paralytic eyelid ectropion. METHODS: Anatomic dissection was performed in 12 fresh cadavers. RESULTS: The data show that the distance between the outer canthus and the crus helicis is 8.733 +/- 1.311 cm. The deep temporal nerve spread radially to the edge of musculus temporalis at 3.852 +/- 0.353 cm and 6.317 +/- 0.330 cm at a 60-degree angle to the horizontal line. The rami of the deep temporal nerve rise at a 60-degree angle to the horizontal line in 3 parts from the outer canthus to the crus helicis. CONCLUSION: In transferring the temporal muscle to treat paralytic ectropion, it is safe to make incisions at the middle one-third between the outer canthus and the crus helicis. The discovery has been used to treat 32 patients with eyelid ectropion of facial palsy, among whom 30(93.8%) got good results. PMID- 11501141 TI - [Diagnosis and treatment of true hermaphroditism: a report of 3 cases and a review of literature]. AB - OBJECTIVE: This paper evaluates the importance of early diagnosis and systemic treatment of true hermaphroditism. METHODS: Three patients with true hermaphroditism, because of inaccurate diagnosis and improper treatment at their early ages, were put in a dilemma when they grew up. After correct diagnosis, they accepted sex determinative operations according to their selective gender role. RESULTS: Through plastic operations, the three patients turned to be a male and two females, with their long time depression being released. CONCLUSION: True hermaphroditism is a rare deformity of ambiguous genitalia characterised by the presence of both ovarian and testicular tissues in the same individual. It is a key point for the patient to form normal sex psyche and adapt society through early precise diagnosis, appropriate gender option and successful operation according to clinical appearances and tissue pathological examination. PMID- 11501142 TI - [The application of external skin expansion in the soft tissue donor site]. AB - OBJECTIVE: To overcome the insufficiency of donor tissue for repairing a large postburn defect. METHODS: The donor site was pre-expanded with an external expansion device in 54 clinical cases. RESULTS: The expansion process was uneventful. The external expansion had delay effect that enabled the flap to be transferred with greater length to width ratio. After external expansion, the donor site of a flap or skin graft could be closed directly. The recipient and donor sites healed simultaneously. CONCLUSION: External expansion is a practical method to provide 'extra' skin for reconstruction of a large skin defect. PMID- 11501143 TI - [Orbital fat preservation and orbicularis muscle flap suspension for lower eyelid pouches]. AB - OBJECTIVE: This is to introduce the experience in the treatment of lower eyelid pouches by orbital fat preservation and orbicularis oculi muscle flap suspension techniques. METHODS: From Sept. 1996 to May 1997, thirty patients with lower eyelid pouches were treated using Hamra's procedure. During the operation, the arcus marginalis was incised. The orbital fat was released and advanced beyond the infraorbital rim and sutured. A lateral-based orbicularis oculi muscle flap was created, and its pedicle was repositioned to the periosteum of the upper lateral orbital rim in a superiomedial direction. RESULTS: A total of 26 patients were available for postoperative follow-up for 3 to 8 months (mean 5.6 months). Of them one patient developed unilateral lower eyelid ectropion due to flabby reposition of the orbicularis oculi muscle flap. The rest obtained excellent cosmetic results without complications. CONCLUSION: This new method has following advantages: 1. The lower eyelid pouches and the infraorbital rim show that may be corrected simultaneously: 2. Sunken eyelids and scleral show that may follow fat removal in conventional lower blepharoplasty can be effectively avoided. 3. "Cheek lift" and a youthful eyelid-cheek complex contour can be obtained. PMID- 11501144 TI - [A clinical and experimental study on autologous nail transplantation ophthalmoplasty]. AB - OBJECTIVE: To seek for better materials for ophthalmoplasty. METHODS: The nails were used as autografts in an experimental and a clinical study. The paws were subcutaneously grafted in the backs of 15 rabbits. The nails were used as the tarsus to constitute the lower eyelid on one patient. On another patient, the toenails were used to restore fracture of the base of the orbit. RESULTS: Without reject reaction, the paws were encapsulated by fibrous tissue ten weeks after grafting. The six patients were followed up for 1 to 5 years and were satisfied with the plastic operations. CONCLUSION: The nail is a practical and an effective autograft in ophthalmoplasty. PMID- 11501145 TI - [Urinary diversion in the one-stage hypospadias repair]. AB - OBJECTIVE: To investigate whether the urinary diversion is helpful for healing of the neourethra. METHODS: Twenty-four cases that had received one-stage urethroplasty were divided into two groups and analyzed retrospectively. The urinary diversion was used in one group of 10 cases. In another group of 14 cases no urinary diversion was used. RESULTS: One case developed a fistula and wound infection in the urinary diversion group. In the no urinary diversion group, 2 out of 5 cases treated with the penile ventrolateral island skin flap urethroplasty developed wound infection and fistulas; excellent results were achieved in all 4 cases treated with the scrotal septal vascular pedicle skin flap urethroplasty; 2 cases developed fistulas and one case had wound infection in the 5 cases treated with the transverse preputial island flap urethroplasty. CONCLUSION: The urinary diversion was not to be used in one-stage urethroplasty using scrotal septal vascular pedicle skin flap. When the neourethra was reconstructed by the penile ventrolateral island skin flap or the transverse preputial island flap, urinary diversion is helpful for spontaneous healing of small fistulas and useful in reducing the incidence of fistulas. PMID- 11501146 TI - [The supporting effects of the pedicle of the subdermal vascular network flap in the rat]. AB - OBJECTIVE: To investigate the role of the pedicle in the survival of the subdermal vascular network skin flap. METHODS: The subdermal vascular network skin flap was created on the back of the rat in the experimental group. The subdermal vascular network skin graft was made at the same position of the rat in the control group. The ratio of length to width of the flap was 4:1. The observation was carried out with naked eyes and instrument. Hair growing on the flap was one of the most important indexes of the observation. RESULTS: It was observed in 116 animals that the proximal 25 percent of the flap possessed regular colour and well-growing hair. CONCLUSION: The pedicle of the subdermal vascular network skin flap in the rat can supply 25 percent of the whole flap. PMID- 11501147 TI - [An anatomical study of the nasolabial fold region]. AB - OBJECTIVE: This study of the nasolabial fold is to provide the morphological basis for maxillofacial plastic and cosmetic surgery. METHODS: Twenty fresh adult cadavers' heads were dissected with microsurgical technique and the structure of the nasolabial region was observed and measured. RESULTS: 1. The data about the length, width and thickness of each mimetic muscle were obtained and presented in the paper. 2. The medial and lateral subcutaneous fat of the nasolabial groove was 1.3 mm and 4.5 mm in thickness respectively. 3. There existed muscle fiber attachments in the medial dermis of nasolabial groove, and in the lateral dermis there were only a few muscle bundle attachments. 4. Among the superficial facial muscles there were not only fasciae but also aponeuroses. The muscles, fasciae and aponeuroses composed a three-dimensional reticular structure. CONCLUSION: This study gave a further support to the theoretical hypothesis of SMAS intermediate tendon. PMID- 11501148 TI - [Advances in the technique of craniofacial surgical computer-assisted system and its clinical applications]. PMID- 11501149 TI - [Study of the uses of growth factors in plastic surgery]. PMID- 11501150 TI - [Gene expression of growth factors and their receptors in healing of partial thickness burn wound in rats]. AB - OBJECTIVE: To investigate the role of growth factors and their receptors in partial-thickness burn wound healing. METHODS: SD rats were used. After 10% total body surface area partial-thickness burn, wound tissues were harvested on postburn days (PBDs) 0(normal control), 1, 3, 5, 7, 10 and 14 respectively. The gene expressions of growth factors and their receptors were determined in wound by in situ hybridization and slot blotting hybridization. At the same time, the process of the wound healing was observed histologically, and the regeneration cycle of epidermal cells and the temporal change in inflammatory cells were measured. RESULTS: Inflammatory cells infiltrated into wound surface were neutrophils, followed by macrophages and lastly lymphocytes. Epidermal cells proliferated most actively on PBD 3 and the mitoses of them increased significantly on day 7 after burn. The gene expression of PDGF, PDGFR and EGFR reached peaks on PBD 1 and the gene expression of EGF and TGF beta-R2 were highest on PBD 3. In addition, the gene expression of TGF beta-R1 and TGF beta 1 increased significantly on PBDs 5 and 7 respectively. CONCLUSION: The data suggested that burn can induce gene expression of EGF, PDGF, TGF-beta 1 and their receptors temporally, spatially, and reversibility, which might play a major role in burn wound healing, and the mutual regulation may exist in the gene expression and the cell cycle. PMID- 11501151 TI - [Effects of hyaluronic acid-stimulating factor on viability and collagen synthesis of fibroblasts]. AB - OBJECTIVE: To study the effects of hyaluronic acid-stimulating factor(HASF) on viability and collagen synthesis of fibroblasts derived from human dermis and scars. METHODS: HASF was purified from amniotic fluid and fetal serum of gestational rabbits, and its activity was determined. To observe the effects of HASF on viability and collagen synthesis of two cell lines, AgNORs staining, 3H proline incorporation and HYP determination were carried out. RESULTS: HASF reduces AgNORs contents in skin FB and scar FBi 3H-proline incorporation and HYP contents were lower in experimental groups than those in controls in both cell lines. Two methods to detect collagen correlate well with each other. CONCLUSION: HASF inhabits viability and collagen synthesis of both cell lines, thus may interfered with the formation of scar during wound healing. PMID- 11501152 TI - [Effects of metallothionein (MT) on burned skin of rats]. AB - OBJECTIVE: To elucidate the relationship between skin viability and lipid peroxidation after deep partial thickness burn injury. METHODS: Deep partial thickness burn was produced. Metallothionin (MT) in different concentrations (0.5 x 10(-6) mol/L, 1 x 10(-5) mol/L) was topically applied. Oxygen consumption, succinate dehydrogenase(SDH), Schiff's base contents and healing time of wound were determined in 24 Wistar rats. Specimens were harvested at 8, 24, and 48-hour postburn. RESULTS: Oxygen consumption and SDH value decreased and Schiff's base value increased after burn injury. With the application of MT, oxygen consumption and SDH value of burned skin were significantly higher compared with control group, and Schiff's base value was significantly lower (P < 0.05 or P < 0.01). The mean healing time of burned wound was two days less in two MT protection groups. CONCLUSION: MT may be beneficial in improving postburn skin viability by attenuating lipid peroxidation of burned tissue. PMID- 11501153 TI - [Experimental study on the early liver injury and expression of TNF-alpha mRNA after burns complicated by endotoxemia]. AB - OBJECTIVE: To observe the distribution and cell localization of TNF-alpha mRNA and its protein and study their role on pathogenesis of liver injury in rats. METHODS: An animal model in which rats were subjected to 20% TBSA III degree burns combined with intraperitoneal injection of lipopolysaccharide (LPS) was used for this experiment. The changes in the hepatic morphology, functions and TNF-alpha content in the serum and the expression, localization of liver TNF alpha and TNF-alpha mRNA were determined with LM and EM, quantitative analysis, immunohistochemistry(IHC) and in situ hybridization (ISH). RESULTS: It was showed that there were sinusoid reaction, KCs activation and degeneration, necrosis of HCs, and platelets aggregation, fibrin deposition and PMNs attachment in sinusoid. The activity of ALT was obviously elevated and the content of ALB was slightly lowered. The contents of TNF-alpha in serum showed the peak values at 3 h. TNF-alpha was mainly localized in sinusoid endothelial cells(SECs), Kupffer cells (KCs) and TNF-alpha mRNA was mainly distributed in KCs, polymorphonuclear neutrophils (PMNs) and macrophages(MPs). CONCLUSION: The results suggest that the expression and localization of TNF-alpha mRNA and its protein are in agreement with the pathological changes of liver injury. TNF-alpha is one of the key cytokines in pathogenesis of liver injury in rats after burns complicated by endotoxemia. PMID- 11501154 TI - [The changes of calcitonin gene-related peptide (CGRP)-containing nerve fibers in adrenal gland of burned rats]. AB - OBJECTIVE: To investigate the changes in calcitonin gene-related peptide(CGRP) within the adrenal glands medulla and cortex of the rat during early post burn period and the effect of CGRP on function of adrenal gland of burned rats. METHODS: The rats were randomly divided into two groups: control and burned. The distribution density of CGRP containing nerve fibers and cells in medulla of adrenal gland were determined at different timepoints post burn. RESULTS: 1. The CGRP-containing nerve fibers were distributed in capsule, cortex and medulla. CGRP-containing nerve fibers were associated with CGRP immunoreactive cell in medulla of adrenal glands. 2. Distribution density of CGRP-containing nerve fiber was decreased, but distribution density of CGRP-containing cells in medulla increased. CONCLUSION: CGRP may affect function of the adrenal gland of burned rats. PMID- 11501155 TI - Inducible overexpression of Bak sensitizes HCC-9204 cells to apoptosis induced by doxorubicin. AB - AIM: To investigate the role of overexpression of Bak in apoptotic pathways and drug susceptibility using doxorubicin and vinorelbine in human HCC-9204 cells. METHODS: An inducible system, MT-II regulatory system which allowed controlled expression of protein upon addition of ZnSO4(100 mumol/L) as an external inducer was used. Stable transfection of pMD-Bak gene was performed on HCC-9204 cells. Apoptotic cells were measured by morphological criteria, as well as by TUNEL assay and flow cytometry. The ability of Bak to decrease clonogenic cell survival was studied by colony-forming assays, while decrease in cell viability was assessed by MTT assay. RESULTS: Cells overexpressing Bak showed extensive cell death with nucleus fragmentation detected by TUNEL assay. FACS analyses showed that Bak could induce significant G1 accumulation and apoptosis in 19.29% cells 24 h after induction. Bak significantly decreased the clonogenic survival following exposure to adriamycin, but not vinorelbine. Furthermore, the time course of cell viability rates following exposure of HCC-9204/Bak cells to adriamycin and vinorelbine was in agreement with the above findings. Bak selectively sensitized HCC-9204 cells to death induced by adriamycin while resisted to vinorelbine. CONCLUSION: Bak may prolong cell cycle in G1 phase, leading to apoptosis and decrease clonogenic survival of HCC-9204 cells in a drug specific manner. PMID- 11501156 TI - Inhibition of experimental cirrhosis in rats by HD-03. AB - AIM: To investigate the protective effect of HD-03 in experimental cirrhosis following chronic intoxication with thioacetamide (TAA). METHODS: The effect of HD-03 (750 mg/kg p.o.) was studied in rats following TAA-induced intoxication (50 mg/kg p.o.) for a period of 90 d. HD-03 was administered as an aqueous suspension. Levels of biochemical markers indicative of hepatotoxicity were assessed in serum and liver. Histopathological evaluation of liver was also carried out to find out the protective effect of HD-03 following TAA-induced chronic intoxication. RESULTS: Administration of TAA at a dose of 50 mg/kg p.o. for 90 d resulted in a significant derangement of serum [serum glutamic pyruvate transaminase (SGPT), serum glutamic oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), albumin and bilirubin] and hepatic (triglycerides, protein, hydroxyproline, collagen and glycogen) biochemical parameters. Histopathological evaluation of liver sections following TAA-intoxication showed necrosis and proliferative changes characteristic of cirrhosis. Simultaneous treatment of TAA intoxicated rats with HD-03 at a dose of 750 mg/kg p.o. for the same duration significantly prevented the changes in both serum and hepatic biochemical parameters. The reversal of serum and hepatic biochemical parameters also correlated with the preservation of liver histoarchitecture in HD-03 treated rats. CONCLUSION: The responses such as membrane stabilization, hepatocellular regeneration, and inhibition of collagen formation are the contributing factors in the correction of TAA-induced cirrhosis by HD-03. PMID- 11501157 TI - Triptolide: a potent inhibitor of NF-kappa B in T-lymphocytes. AB - AIM: To study the effect of triptolide on the activity of nuclear factor-kappa B (NF-kappa B) and gene expression of I kappa B alpha, a major inhibitor of NF kappa B, in human Jurkat T cell line. METHODS: Jurkat cells treated with or without PMA/PHA were incubated with varied doses of triptolide for different time periods. The activity of NF-kappa B in Jurkat cells was measured with electrophoretic mobility shift assays (EMSA), and the mRNA expression of I kappa B alpha in Jurkat cells was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: PMA/PHA treatment was found to induce NF-kappa B activation rapidly. Triptolide treatment decreased the activity of NF-kappa B in both PMA/PHA treated and untreated Jurkat cells. This effect of triptolide was in a dose-dependent manner and was more obvious in cells treated with PMA/PHA. The mRNA expression of I kappa B alpha were upregulated by triptolide, while this effect was more potent in cells without PMA/PHA treatment. CONCLUSION: Triptolide is a potent inhibitor of NF-kappa B activation in T lymphocyte and this effect is partly due to the upregulation of I kappa B alpha mRNA expression. PMID- 11501158 TI - Effect of protein kinase C on guinea pig sperm acrosome reaction induced by progesterone. AB - AIM: To investigate the effect of protein kinase C (PKC) on the guinea pig sperm acrosome reaction (AR) induced by progesterone (P4). METHODS: Guinea pig spermatozoa were preincubated in MCM-Ca2+ DF medium at 38.5 degrees C under 5% CO2/air for 6-6.5 h for capacitation, and were then washed through a three-step gradient Percoll, resuspended in MCM-Ca2+ DF and adjusted to 5 x 10(9) cells/L. The AR was assessed by means of phase-contrast microscopy (expressed as percent of AR) 15 min after different reagents were added. RESULTS: In the presence of Ca2+ 2 mmol/L, phorbol 12,13-dibutyrate (PDB) or the permanent synthetic diacylglycerol, 1-oleoyl-2-acetyl-sn-glycerol (OAG) initiated the AR of spermatozoa in a concentration-related manner, whereas staurosporine (STA) exhibited an inhibition of the AR. PDB and OAG significantly enhanced P4-induced AR that was greater than that treated with either agonist separately. STA markedly inhibited the P4-induced AR. The AR induced by PDB was completely antagonized by inclusion of egtazic acid. Also, both egtazic acid and nifedipine antagonized the AR induced by P4 and PDB + P4. In addition, neomycin also prevented the P4-induced AR. This effect could be partly reversed by PDB. CONCLUSION: Activation or inhibition of PKC significantly plays an important role in the signal transduction pathway for the physiological AR. PMID- 11501159 TI - Metabolism of ginsenoside Rg1 by intestinal bacteria. II. Immunological activity of ginsenoside Rg1 and Rh1. AB - AIM: To compare the effect of ginsenoside Rg1 and its metabolite Rh1 on proinflammatory cytokines and their mRNA expression by THP-1 cells. METHODS: Human peripheral blood mononuclear cells (PBMC) were incubated with Rg1 and Rh1 at concentrations of 0.1, 1, 10, and 100 mg/L, and the cell proliferation was measured 24 h after incubation. Radioimmunoassay (RIA) was used to detect the production of proinflammatory cytokines, TNF alpha, IL-1 alpha, and IL-8. TNF alpha mRNA level was detected by reverse transcription polymerase chain reaction (RT-PCR) after administration of Rg1 and Rh1. RESULTS: Rg1 and Rh1 (at concentration of 0.1, 1, 10, 100 mg/L) had no effect on PBMC proliferation. Rh1 1 mg/L could upregulate the productions of TNF (and IL-8 induced by lipopolysaccharides (LPS) 10 mg/L plus phorbol myristate acetate (PMA) 200 nmol/L, however, Rg1 showed an inhibitory effect on TNF alpha production induced by LPS 100 mg/L. Rg1 1 mg/L and Rh1 100 mg/L enhanced the production of IL-1 alpha level in THP-1 cells in the presence of LPS 10 mg/L. RT-PCR revealed that Rh1 stimulated TNF alpha mRNA expression in suitable stimulatory conditions. CONCLUSION: Rg1 and Rh1 have different effects on the production of cytokines produced THP-1 cells stimulated by LPS and PMA. PMID- 11501160 TI - Effects of rhM-CSF expressed in silkworm on cytokine productions and membrane molecule expressions of human monocytes. AB - AIM: To study the effects of recombinant human macrophage colony-stimulating factor (rhM-CSF) expressed in silkworm on cytokine productions and membrane molecule expressions of monocytes. METHODS: The rhM-CSF was added to the human peripheral blood monocyte cultures and 3 d later, the culture supernatants and cells were collected, respectively. TNF-alpha, IL-6, IL-8, and IFN-alpha levels in the supernatants were detected by biological activity test or ELISA and expressions of CD11b, CD16, HLA I, and HLA II on the cellular surface were examined by the method of alkaline phosphatase anti-alkaline phosphatase complex (APAAP). RESULTS: The rhM-CSF promoted TNF-alpha, IL-6, and IL-8 inductions of monocytes and increased the percentages of CD11b, CD16, HLA I, and HLA II molecule expression on monocytes. CONCLUSION: The rhM-CSF plays a role in monocyte function up-regulation and has a certain practical value in immunological therapy. PMID- 11501161 TI - Influence of AVP4-8 on MAPK activity in astrocytic glial and fetal neuronal cells in primary culture. AB - AIM: To study the effect of argipressin (4-8) (AVP4-8) on the mitogen-activated protein kinase (MAPK) activity in astroglial culture and fetal neuronal culture from rat cerebral cortex and hippocampus. Some protein kinases involved in this signal pathway were also addressed. METHODS: Rat brain primary cells were cultured in serum free medium or starved for 24 h before use. Cells were transferred to Ca2+ and Mg2+ free Dulbeco's phosphate buffer (D-PBS) with various drugs. MAPK activity was measured. RESULTS: The main findings were: (1) AVP4-8 induced the MAPK activity in rat brain astroglial culture but not in fetal neuronal cultures. And this was blocked by ZDC (C) PR, an antagonist of AVP4-8. (2) PD98059, a potent selective inhibitor of MAPK/ERK kinase (MEK) and GF109203X, a specific inhibitor of protein kinase C (PKC) abolished AVP4-8-evoked MAPK activity on astrocytes. CONCLUSION: AVP4-8 can activate the MAPK activity in astrocytes but not in fetal neuronal culture. MEK and PKC may be involved in the AVP4-8-evoked cascade. PMID- 11501163 TI - Facilitating effects of histamine on spatial memory deficit induced by scopolamine in rats. AB - AIM: To investigate whether or not histamine was involved in scopolamine-induced spatial memory deficits evaluated in 8-arm radial maze performance of rats. METHODS: Eight-Arm radial maze performance was used to measure spatial memory in rats, and the brain regions were subsequently dissected and histamine contents were determined by HPLC. RESULTS: Intracerebroventricular (icv) injection of histamine (100 or 200 ng) or thioperamide (50 micrograms), and intraperitoneal (i.p.) injection of histidine (1000 mg/kg) ameliorated memory impairment induced by scopolamine regarding both parameters of radial maze performance. 2 Thiazolylethylamine, but not 4-methylhistamine showed the similar effect to histamine. Both histamine (200 ng, icv) and histidine (1000 mg/kg, i.p.) were equally effective in increasing the histamine content in the cortex, hippocampus, and hypothalamus. CONCLUSION: These results suggest that brain histamine plays an important role in learning and memory, and its action may be due to cholinergic neurons. PMID- 11501162 TI - Adenovirus-mediated expression of human secretor type alpha(1,2) fucosyltransferase reduces level of Gal alpha(1,3)Gal epitope. AB - AIM: To test the potential of human secretor type alpha(1, 2) fucosyltransferase [Se alpha(1,2)FT] to downregulate the expression of Gal alpha(1,3)Gal epitope (gal epitope) in cultured cell lines. METHODS: Expression of Se alpha(1,2) FT was mediated by human adenoviral vector. Flow cytometric analysis was used to compare the expression level of H blood group antigen or gal epitope. MTT was employed to assess the susceptibility of mouse NIH3T3 cells to human natural antibody and complement mediated lysis. RESULTS: A recombinant replication-deficient adenovirus (rAdv) containing human Se alpha(1,2)FT cDNA (Ad5hSeFT) was designed and successfully constructed. Flow cytometric analysis showed that after mock infection, Ad5null infection, and Ad5hSeFT infection, the mean fluorescence intensity (MFI) values for the binding of Ulex europaeus I (UEA-I) lectin to NIH3T3 cells were 2.3 +/- 0.6, 2.1 +/- 1.0, and 36.5 +/- 5.9, respectively; MFI values for the binding of Griffonia simplicifolia isolectin B4 (GS-IB4) lectin to NIH3T3 cells were 167 +/- 23, 170 +/- 19, and 100 +/- 14, respectively; MFI values for the binding of human natural IgG and IgM antibodies to NIH3T3 cells were 31 +/- 3, 32 +/- 4, and 22 +/- 4, respectively. CONCLUSION: H blood group antigen was detected on NIH3T3 cells after Ad5hSeFT infection and resulted in more than 40% reduction in the level of gal epitope on the cell surface. This reduction increased the resistance of NIH3T3 cells to lysis by normal human serum. PMID- 11501164 TI - Alterations of amino acid levels from striatum, hippocampus, and cerebral cortex induced by global cerebral ischemia in gerbil. AB - AIM: To investigate global cerebral ischemia-induced alterations in the levels of glutamate, aspartate, gamma-aminobutyric acid (GABA), glutamine, glycine, and taurine from hippocampus, striatum, and cerebral cortex in gerbils. METHODS: The gerbil global cerebral ischemia model was prepared by bilateral carotid artery occlusion; the contents of amino acids were assayed using high performance liquid chromatography (HPLC) combined with fluorescent detection after precolumn derivatization. RESULTS: After the ligation of bilateral carotid artery for 5 min and reperfusion for 60 min, the contents of glutamate from hippocampus, striatum, and cortex in gerbils were increased by 40%, 49%, and 67%, respectively. Similarly, the global cerebral ischemia resulted in increase by 80%, 69%, and 83% of aspartate contents in hippocampus, striatum, and cortex, respectively. Moreover, the same treatment also induced significant increases in the contents of GABA, glutamine, glycine, and taurine from various brain regions in gerbils. Furthermore, pretreatment with ketamine (120 mg/kg, i.p.) reversed ischemia evoked increases of glutamate, aspartate, glycine, and glutamine in hippocampus, striatum, and cortex of gerbils. However, administration of ketamine (120 mg/kg, i.p.) markedly suppressed but not abolished the ischemia-induced increases of taurine and GABA from hippocampus, striatum, and cortex in gerbils. CONCLUSION: The increases of glutamate, aspartate, glycine, and glutamine induced by acute global cerebral ischemia may constitute the biochemical basis of ischemic brain damage. Correspondingly, the release of GABA and taurine may be an important self protective mechanism. Ketamine may protect neurons against ischemic insult by inhibiting global cerebral ischemia-evoked increase of glutamate, glycine, and aspartate. PMID- 11501165 TI - Effects of MCI-154, a calcium sensitizer, on cardiac function in endotoxemic rabbits. AB - AIM: To observe the effects of MCI-154, a calcium sensitizer, on cardiac function after endotoxic shock. METHODS: The rabbits were intravenously injected with MCI 154 0.1 mg.kg-1 at 10 h after the administration of endotoxin 1.0 mg.kg-1, followed by a continuous infusion of normal saline (NS) 50 mL.kg-1 + MCI-154 0.1 mg.kg-1. During this process, the parameters of cardiac function were measured. RESULTS: Ten hours after the endotoxin injection, heart rate (HR) increased noticeably while the mean arterial blood pressure (MAP), left ventricular systolic pressure (LVSP), isovolumetric pressure (IP), myocardial contractility (MC), and the area of p-dp/dtmax vector loop (Lo) were all markedly decreased. Treatment with NS 50 mL.kg-1 alone had slight effects on these parameters. LVSP, IP, MC, and Lo were all markedly increased while HR did not obviously change and left ventricular end-diastolic pressure (LVEDP) was reduced markedly following MCI-154 administration in endotoxic shock rabbits. The parameters of MC were improved nearly to the same values as in sham shock group and were markedly higher than in NS treated group. CONCLUSION: MCI-154 exerts remarkable therapeutic effects on cardiac dysfunction after endotoxic shock. PMID- 11501166 TI - Effect of MCI-154, a calcium sensitizer, on calcium sensitivity of myocardial contractile system in endotoxemic rats. AB - AIM: To investigate the effect of MCI-154 on calcium sensitivity of myocardial contractile system in endotoxemic rats. METHODS: Skinned right ventricular papillary muscles from endotoxemic rats were prepared by saponin 500 mg/L. Forces of the skinned muscles were recorded when they were activated sequentially by different concentrations of Ca2+ with or without cardiotonic agents. The tension pCa relationship and pCa50 of the skinned fibers were taken as the index of Ca2+ sensitivity of myocardial contractile system. RESULTS: The maximal Ca(2+) activated tension (Tmax) was lower, and pCa50 was reduced in endotoxemia group as compared with those in sham control group. Milrinone 50 mumol/L could not counteract the above abnormalities. However, after MCI-154 10 mumol/L was added, the Tmax and pCa50 were increased to an extent similar to that of sham control group and significantly higher than those of endotoxemia group and endotoxemia + milrinone group. Furthermore, such effects of MCI-154 were concentration dependent. CONCLUSION: The Ca2+ sensitivity of cardiac contractile system in endotoxemic rats is decreased. MCI-154 can reverse the decreased sensitivity and increase Tmax of myocardial muscles from endotoxemic rats. PMID- 11501167 TI - Inhibitory effects of purified green tea epicatechins on contraction and proliferation of arterial smooth muscle cells. AB - AIM: The present study was aimed to examine the vasorelaxant and antiproliferative responses to purified green tea epicatechin mixture, ( )epicatechin and (-)epigallocatechin gallate on rat arterial smooth muscle cells. METHODS: Changes in isometric force were measured by Grass force transducer and effects on cell proliferation were evaluated using [3H]thymidine incorporation assay. RESULTS: Epicatechin mixture, (-)epicatechin and (-)epigallocatechin gallate, which we isolated and purified from jasmine green tea, concentration dependently, reduced the contractile response to phenylephrine in rat isolated aortic rings with (-)epigallocatechin gallate being more effective. These three agents also inhibited [3H]thymidine incorporation into DNA in cultured rat aortic smooth muscle cells in a concentration-dependent manner. CONCLUSION: The purified epicatechin derivatives from jasmine green tea relaxed the isolated rat arteries preconstricted by phenylephrine and inhibited aortic smooth muscle cell proliferation. PMID- 11501168 TI - Effect of melittin on potassium currents and action potential in ventricular myocytes of guinea pig. AB - AIM: To examine the effects of melittin (Mel), the major component of bee venom, on delayed rectifier K+ current (Ik), inward rectifier K+ current (Ikl) and action potential (AP) in guinea pig ventricular myocytes. METHODS: Ik, Ikl, and AP were recorded using the whole-cell patch-clamp technique. RESULTS: The action potential duration (APD) was shortened by Mel in a concentration-dependent manner. Mel 0.05, 0.1, 0.2 mumol.L-1 shortened APD50 from (520 +/- 55) to (459 +/ 91) (n = 5, P > 0.05), (385 +/- 102) (n = 5, P < 0.01), and (281 +/- 81) ms (n = 5, P < 0.01), respectively; and APD90 from (613 +/- 96) to (536 +/- 93) (n = 5, P > 0.05), (467 +/- 96) ms (n = 5, P < 0.01), and (354 +/- 95) ms (n = 5, P < 0.01), respectively. Mel increased the amplitude of Ik also in a concentration dependent manner. Mel 0.05, 0.1, 0.2 mumol.L-1 increased Ik from (295 +/- 109) to (371 +/- 142) (n = 5, P < 0.05), (467 +/- 180) (n = 5, P < 0.05), (552 +/- 248) pA (n = 5, P < 0.05), respectively at testing potential of + 40 mV. But no significant effect of Mel on Ikl was observed at these concentrations. CONCLUSION: Mel significantly increased Ik in a concentration-dependent manner which contributed to shortening of APD. PMID- 11501169 TI - Hypoxic preconditioning upregulates KATP channels through activation of protein kinase C in rat ventricular myocytes. AB - AIM: To test: 1) if hypoxic preconditioning upregulates the activity of KATP channels in isolated adult rat cardiac myocytes; 2) if the upregulation involves protein kinase C (PKC). METHODS: By the whole-cell patch-clamp recording technique, KATP channel currents (IKATP) were measured in the cardiomyocytes with no pretreatment (CON), hypoxic exposure preceding reoxygenation (HPC), phorbol 12 myristate 13-acetate (PMA), and chelerythrine addition with HPC (CH + HPC), respectively. RESULTS: At 0 mV and 5 min after 2, 4-dinitrophenol (DNP) perfusion, the membrane currents for the CON, HPC, PMA, and CH + HPC were (3.5 +/ 1.9), (7.7 +/- 1.5), (7.5 +/- 3.3), (4.6 +/- 2.4) nA, respectively. Compared with the CON, the IKATP in the HPC and PMA were significantly augmented (P < 0.01) while the IKATP in the CH + HPC remained similar to the CON (P > 0.05). CONCLUSION: 1) hypoxic preconditioning stimulated the activity of PKC and markedly enhanced the activity of KATP channels in the isolated rat cardiac myocytes; 2) PKC activation was involved in the upregulation of KATP channels. PMID- 11501170 TI - Effect of valsartan and fosinopril on catecholamine-induced cardiac hypertrophy. AB - AIM: To study the influence of angiotensin (Ang) II receptor antagonist (AT1) valsartan and angiotensin-converting enzyme (ACE) inhibitor fosinopril on the cardiac hypertrophy induced by catecholamine. METHODS: A cardiac hypertrophy model was produced by i.p. norepinephrine (NE) 1.5 mg.kg-1.d-1 x 15 d. The animals were divided into four groups: 1) control (sodium chloride), 2) NE, 3) NE + fosinopril, 4) NE + valsartan. Fosinopril ig 15 mg.kg-1.d-1 x 15 d, valsartan ig 30 mg.kg-1.d-1 x 15 d. RESULTS: Valsartan ig 30 mg.kg-1.d-1 x 15 d and fosinopril ig 15 mg.kg-1.d-1 x 15 d prevented left ventricular hypertrophy induced by NE and decreased the content of collagen in myocardium; valsartan and fosinopril both elevated the myosin ATPase activity, Na+, K(+)-ATPase activity in membrane, and Ca(2+)-ATPase activity in mitochondrias. Apoptosis was induced in cardiomyocytes by catecholamine. Valsartan and fosinopril both inhibited apoptosis, and no significant differences were found in the apoptotic index between the two treatment groups. CONCLUSION: Valsartan and fosinopril prevent the remodeling of cardiac hypertrophy induced by norepinephrine. Cardiac myocyte apoptosis may play a key role in the heart remodeling. PMID- 11501171 TI - Free radical scavenging and inhibition of lipid peroxidation by magnesium lithospermate B. AB - AIM: To study the effect of magnesium lithospermate B (MLB) on the lipid peroxidation and on its free radical scavenging activity. METHODS: MLB was incubated in rat tissue homogenate or in a free radical generating system. MLB induced inhibition of lipid peroxidation and its scavenging activity on superoxide anions and hydroxyl radicals was studied using colorimetric estimation. RESULTS: MLB inhibited the lipid peroxidation induced by either an auto-oxidant or Fe2+/VitC in vitro, in the liver homogenate, the inhibitory rate of MLB (10 mg/L) being 69.2% and 57.7%, respectively. MLB (25 and 50 mg/kg) decreased the amount of thiobarbituric acid reactive substances (TBARS) in rat serum, liver, kidney, and heart. However, it did not inhibit the lipid peroxidation of brain homogenate ex vivo. MLB scavenged superoxide anions generated from xanthine/xanthine oxidase system and iron-dependent hydroxyl radicals. CONCLUSION: MLB is an inhibitor of lipid peroxidation and scavenge superoxide anions and hydroxyl radicals both in vitro and ex vivo. PMID- 11501172 TI - Effects of magnesium lithospermate B on aggregation and 5-HT release in rabbit washed platelets. AB - AIM: To study the effects and mechanism of magnesium lithospermate B(MLB) on rabbit platelet aggregation and 5-HT release. METHODS: The platelet aggregation was determined by Born's method. Release of serotonin (5-HT) and formation of thromboxane A2 (TXA2) were measured by fluorophotometry and radioimmunoassay (RIA) respectively. Cytoplasmic free Ca2+ concentration ([Ca2+]i) in platelets was measured by Fura 2-AM fluorescence technique. RESULTS: In washed platelets, thrombin (200 U/L) or arachidonic acid (AA) (30 mumol/L)-induced aggregation was inhibited by MLB 50-800 mg/L in a concentration-dependent manner. In addition, MLB had more inhibitory effects on platelet aggregation in the absence of extracellular calcium with IC50 of 102 mg/L than in the presence of CaCl2 1 mmol/L with IC50 of 194 mg/L. MLB concentration-dependently decreased the thrombin-activated release of 5-HT, whereas it did not affect the formation of TXA2 in platelets. Furthermore, MLB not only inhibited the rise of [Ca2+]i in thrombin stimulated platelets, but decreased the [Ca2+]i in resting platelets. CONCLUSION: MLB inhibited the aggregation and 5-HT release in rabbit platelets and it is probably by attenuating intracellular calcium concentration. PMID- 11501173 TI - Polymorphism of CYP450 and cancer susceptibility. AB - CYP450s form a superfamily involved in the metabolism of many endogenous and exogenous substrates. In this review, the substrates, probe drugs, genetic polymorphisms, and associated cancers of CYP1A1, CYP1A2, CYP2C19, CYP2D6, CYP2E1, and CYP3A4 are summarized. PMID- 11501174 TI - Gender difference in letrozole pharmacokinetics in rats. AB - AIM: To study gender difference in letrozole (Letr) pharmacokinetics in rats. METHODS: Letr concentrations in plasma and tissues were determined after ig administration of Letr 2 mg/kg. Recoveries of Letr in urine and feces were also analyzed. RESULTS: Marked gender differences were found 6 h after ig Letr 2 mg/kg, the plasma concentrations of Letr in male rats were significantly (P < 0.01) lower than those in female rats. For example, at 24, 36, 48, and 72 h after administration, plasma concentrations in female rats were about 3.3, 5.6, 10.5, and 7.4-fold of that of male rats, respectively. AUC value of Letr in male was only about one-third of that in female rats. Estimated terminal phase half-lives (T1/2) were 10.5 and 40.4 h, respectively. In female rats, cumulative excreted fractions of Letr in urine and feces were 5.8% +/- 1.4% and 6.6% +/- 1.1% within 120 h after administration, respectively, but in male rats, the excreted fractions of Letr in urine and feces were only 1.30% +/- 0.59% and 0.87% +/- 0.31%. Letr concentrations in female rat tissues were significantly (P < 0.01) higher than those in male rat tissues 24 h after administration. CONCLUSION: There are marked gender differences in Letr pharmacokinetics in rats. PMID- 11501175 TI - Interaction of combined administration of intrathecal morphine with subcutaneous morphine or buprenorphine. AB - AIM: To analyze the mode of interaction of combined administration of intrathecal morphine with subcutaneous morphine or buprenorphine. METHODS: Different groups of rats were scheduled to undergo administration of intrathecal (ith) morphine, subcutaneous (s.c.) morphine, s.c. buprenorphine, and the combinations of ith morphine with s.c. morphine or buprenorphine in a series of dose ratios. Nociceptive responses of hind paws of each animal were measured by means of "plantar stimulation" test. The test latency was converted to the percent of maximal possible effect (% MPE). RESULT: Morphine ith, morphine s.c., buprenorphine s.c., as well as combinations in all dose ratios increased the % MPE in a dose-dependent manner. Isobolograms showed that the ED50 points determined for the combinations were plotted significantly left to the theoretical additive line. CONCLUSION: The combination of morphine ith with either morphine s.c. or buprenorphine s.c. resulted in a synergistic effect. This interaction might be due to the activation of the synergistic antinociceptive mechanisms between supraspinal and spinal levels. PMID- 11501176 TI - Enzyme kinetics and inhibition of nimodipine metabolism in human liver microsomes. AB - AIM: To study the enzyme kinetics of nimodipine (NDP) metabolism and the effects of selective cytochrome P-450 (CYP-450) inhibitors on the metabolism of NDP in human liver microsomes in vitro. METHODS: Microsomes from six individual human liver specimens were used to perform enzyme kinetic studies and the kinetic parameters were estimated by Eadie-Hofstee equation. Various selective CYP-450 inhibitors were used to investigate their effects on the metabolism of NDP and the principal CYP-450 isoform involved in dehydrogenation of dihydropyridine ring of NDP in human liver microsomes. RESULTS: There was an important intersubject variability in NDP metabolism in human liver microsomes. For NDP dehydrogenase activity, the Km value was (36 +/- 11) mumol and the Vm value was (17 +/- 7) mumol.g-1.min-1. The dehydrogenation of dihydropyridine ring of NDP was competitively inhibited by ketoconazole (Ket) and troleandomycin (TAO), and the Ki values for Ket and TAO were 0.59 and 122.2 mumol, respectively. Phenacetin (Pnt), quinidine (Qui), diethyldithiocarbamate (DDC), sulfaphenazole (Sul), and tranylcypromine (Tra) had a little or no inhibitory effects on the dehydrogenation of NDP. CONCLUSION: The intersubject variability of NDP pharmacokinetics was attributed to the metabolic polymorphism of NDP in liver. Cytochrome P-4503A (CYP3A) is involved in the dehydrogenation of dihydropyridine ring of NDP. PMID- 11501177 TI - Mechanisms of regulation of tyrosine phosphorylation of NMDA receptor subunit 2B after cerebral ischemia/reperfusion. AB - AIM: To study the mechanisms of the regulation of the tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor subunit 2B(NR2B) in the gerbil hippocampal synaptosomes following ischemia/reperfusion (I/R). METHODS: Transient (15 min) cerebral ischemia was produced by bilateral carotid artery occlusion procedure. The tyrosine phosphorylation of NR2B was analyzed by immunoprecipitation and immunoblot assay. RESULTS: Transient forebrain ischemia for 15 min caused a marked decrease in the levels of tyrosine phosphorylation of many protein bands including 180 kDa protein. Transient ischemia followed by reperfusion induced rapid (within 15 min of reperfusion), and sustained (for at least 48 h) increase in the tyrosine phosphorylation of many protein bands including 180 kDa protein. Immunoprecipitation and immunoblot confirmed that NR2B is among the phosphorylated 180 kDa protein. Maximal phosphorylation of 180 kDa band corresponding to NR2B (1.8 fold relative to sham-operated controls) was reached at 6 h of reperfusion following 15 min of cerebral ischemia. But the level of protein expression of NR2B did not change. Administration of ketamine (KT), a non competitive NMDA receptor antagonist, or nifedipine (ND), an L-type voltage gated calcium channel (L-type VGCC) blocker, 20 min before ischemia attenuated stimulation of the tyrosine phosphorylation of NR2B without affecting the level of protein expression of NR2B. Under these conditions, non-NMDA receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX) had no effect on the level of tyrosine phosphorylation. Protein tyrosine phosphatase (PTP) inhibitor vanadate and protein tyrosine kinase (PTK) inhibitor genestein resulted in the increase and the decrease of the tyrosine phosphorylation of NR2B, respectively. Src coprecipitated with NR2B protein. CONCLUSION: The increase of the tyrosine phosphorylation of NR2B induced by I/R has relation to NR and L-type VGCC; PTK and PTP participate in the regulation of the tyrosine phosphorylation of NR2B during I/R. Src that associates with NR2B may play an important role in the regulation of the tyrosine phosphorylation of NR2B during I/R. PMID- 11501178 TI - Building 3D-structural model of kappa opioid receptor and studying its interaction mechanism with dynorphin A(1-8). AB - AIM: To construct the 3D-structural model of human kappa opioid receptor (HKOR) and study its interacting mechanism with dynorphin A(1-8) (Dyn8). METHODS: Comparative molecular modeling was applied to build the 7 transmembrane (TM) helical domain of HKOR using the bovine rhodopsin (OPSD) model as a template. Molecular dynamics was performed to minimize the HKOR model and to simulate the 3D-structure of Dyn8 based on the NMR results of dynorphin A(1-14). The extracellular loops (EL) were built by self-constructed database searching. DOCK4.0 program was performed to construct Dyn8 complex with HKOR. RESULTS: (1) The model of HKOR was obtained and validated by theoretical and experimental data. (2) The Dyn8-HKOR interacting mechanism is reasonably explained: Side chain of residue Asp138 interacts with protonated nitrogen atom at the N-terminal residues of Dyn8 through electrostatic and hydrogen bonding, which play an important role in ligand binding with receptor. (3) Negatively charged amino acids in the second extracellular loop (EL2) as Asp223 and Glu209 interact with the C-terminal positively charged residues in Dyn8, and Glu209 is a likely determinant of peptide ligand specificity. CONCLUSION: Some amino acid residues positioned in EL2, TM3, TM4, and TM5 form the binding site and therefore determine the selectivity of kappa peptide agonist. PMID- 11501179 TI - Inhibition of calcium signaling in terminal and soma of carp retinal bipolar cells by GABA. AB - AIM: To investigate the effect of activation of gamma-aminobutyric acid (GABA) receptors on high K(+)-evoked Ca2+ signaling in the terminal and soma of carp retinal ON-type bipolar cells. METHODS: Freshly dissociated carp retinal cells were loaded with fluo-3AM and then the fluorescence measurements were performed on a confocal laser-scanning microscope. RESULTS: Ca2+ signaling evoked by high K+ 35 mmol/L was completely suppressed in both the terminal and soma of bipolar cells by GABA 100 mumol/L. However, different results were found in the terminal and soma when only one subtype of GABA receptors was activated. While activation of either GABAA or GABAC receptors totally suppressed Ca2+ signaling in the soma, a gradual elevation of [Ca2+]i appeared in the terminal. GABA 10 mumol/L could also completely suppress Ca2+ signaling in the soma, but could only partially reduce Ca2+ signaling in the terminal. CONCLUSION: Activation of both GABAA and GABAC receptors could completely inhibit high K(+)-evoked Ca2+ signaling in the terminal and soma of carp retinal ON-type bipolar cells. While activation of either GABAA or GABAC receptors alone still totally suppressed Ca2+ signaling in the soma, a gradual elevation of [Ca2+]i appeared in the terminal, which may be due to desensitization of GABA receptors. PMID- 11501180 TI - Changes and mechanisms of protein-tyrosine kinase and protein-tyrosine phosphatase activities after brain ischemia/reperfusion. AB - AIM: To study the changes and mechanisms of protein-tyrosine kinase (PTK) and protein-tyrosine phosphatase (PTP) activities in the hippocamal synaptosome following cerebral ischemia/reperfusion (I/R) in gerbil. METHODS: Transient (15 min) global ischemia was produced by bilateral carotid artery occlusion. Total PTK and PTP activities were measured by [r-32P] incorporation and colorimetric analysis, respectively. Src and proline-rich tyrosine kinase2 (PYK2) activities were measured by immunoprecipitation and [r-32P] incorporation. RESULTS: Total PTK activity increased significantly after I/R, but the PTP activity did not change. The Src activity was much higher than PYK2 activity in sham-operated controls. I/R mainly caused a pronounced increase in Src activity, but not PYK2 activity. The increase in Src activity had no relation to the expression of Src protein. Administration of ketamine (KT) or nifedipine (ND) 20 min before ischemia caused a decrease in total PTK and Src activities, and no change in the PYK2 and PTP activities. CONCLUSION: The increase in PTK activity caused by I/R may be mainly due to the increase in Src activity. This increase in Src activity has no relation to the expression of Src protein. But it is related to the activation of NMDA (N-methyl-D-aspartate) receptor (NR) and L-type voltage-gated calcium channel (L-type VGCC). In other words, the increase in total PTK and Src activities induced by I/R may be mediated via NR and L-type VGCC. The PTP activity did not change during I/R. PMID- 11501181 TI - Effects of salvianolic acid-A on rat hepatic stellate cell proliferation and collagen production in culture. AB - AIM: To investigate the effects of salvionolic acid-A (SA-A), one of main effective components of Salvia miltiorrhiza for its antifibrotic action, on the cell proliferation and collagen production in cultured hepatic stellate cells (HSC). METHODS: HSC were isolated through in situ perfusion of liver with pronase E and collagenase, and gradient centrifugation with Nycodenz. The cultured HSC were incubated with SA-A 0.1-100 mumol/L for 24 h. MTT spectrometric assay and intercellular incorporation of methyl-[3H]thymidine ([3H]TdR) was used to assess the cell proliferation. The amount of collagen was semi-quantified by ponceau staining and image analysis, the amount of type I collagen secretion was measured with ELISA and normalized by the total protein of cell layer. The total RNA was prepared from the control cells and the drug treated cells respectively, and the expression of pro-collagen alpha 2 (I) mRNA was semi-quantitatively analyzed with RT-PCR. RESULTS: SA-A 100 mumol/L showed a little cytotoxity, SA-A 0.1-10 mumol/L did not influence cell morphology, and SA-A 1-100 mumol/L decreased the cell proliferation significantly in a concentration-dependent manner (P < 0.05). SA-A 1, 10, 100 mumol/L decreased the cell collagen deposition by 78.6%, 71.8%, and 61.3% of the control respectively (P < 0.05), and decreased type I collagen secretion to 53.1%, 52.6%, and 49.5% (P < 0.01 or P < 0.05). Both SA-A 1 and 10 mumol/L downregulated procollagen alpha 2 (I) mRNA expression remarkably (P < 0.05). CONCLUSION: SA-A inhibited HSC proliferation and collagen expression. The inhibitory effect on HSC activation is the main mechanism of SA-A action against liver fibrosis. PMID- 11501182 TI - Antihypertensive effects of D-polymannuronic sulfate and its related mechanisms in renovascular hypertensive rats. AB - AIM: To investigate the antihypertensive effects of D-polymannuronic sulfate (DPS), a kind of sulfated polysaccharide, and the underlying mechanisms in renovascular hypertensive rats (RHR). METHODS: Used two-kidney one clip (Goldblatt, 2-K 1C) method to produce RHR model. DPS was given i.v. or ig for 5 wk with the initiation of establishment of RHR. Serum nitric oxide (NO) was determined with NO kit; plasma angiotensin II (Ang II) and endothelin-1 (ET-1) were measured by radioimmumoassays. RESULTS: In acute therapeutic experiments, DPS markedly reduced systolic blood pressure (SBP) and diastolic blood pressure (DBP) dose-dependently and decreased heart rate (HR) with reduction in arterial blood pressure. In the prophylactic experiments, DPS prevented the rise in SBP and DBP in a dose-dependent manner. The hypotensive potency of DPS 50 mg/kg is comparable to that of captopril (14 mg/kg). Moreover, DPS elevated serum NO contents and lowered plasma concentrations of Ang II and ET-1. CONCLUSION: The antihypertensive activities of DPS might be involved both in increasing the generation of nitric oxide and in decreasing the production of angiotensin II and endothelin-1 in vivo. PMID- 11501183 TI - Rapid assay of A1c-type glycosylated hemoglobin in blood of diabetic rats using fast protein liquid chromatography. AB - AIM: To develop a fast protein liquid chromatography (FPLC) method for the assay of Alc-type glycosylated hemoglobin in diabetic rats. METHODS: Venous blood was collected from rats. After the erythrocytes were washed and sedimented, the hemolysate was prepared and diluted with isotonic saline as a sample for the assay. Samples were then separated on Mono_S_HR_5/5 cation exchange column by a lithium chloride gradient elution system. The Alc-type glycosylated hemoglobin was monitored by measuring the absorbance at 415 nm. RESULTS: The Alc-type glycosylated hemoglobin was well separated from total hemoglobin. The average HbAlc amount in diabetic rats was determined to be (3.6% +/- 0.6%, n = 7), while the corresponding figure in normal rats was (1.4% +/- 0.4%, n = 7, P < 0.01). CONCLUSION: This FPLC method is easy, rapid and reproducible, and can be used for an assay of diabetic rat HbAlc to evaluate and screen new drugs for diabetes mellitus therapy. PMID- 11501184 TI - Inhibitory effect of disodium quercetin-7,4'-disulfate on aggregation of pig platelets induced by thrombin and its mechanism. AB - AIM: To study the inhibitory effect of semi-synthesized quercetin derivatives- disodium quercetin-7,4'-disulfate (DQD) on the platelet aggregation induced by thrombin and its mechanism. METHODS: Platelet aggregation was analysed by turbidimetry. Cytosolic free calcium concentration ([Ca2+]i) was determined by Fura-2 fluorescence technique. Activity of Ca2+/PL dependent protein kinase C (PKC) was assayed by incubating PKC with histone III S and [gamma-32P]ATP. The cytoskeletal proteins were precipitated by Triton and separated by SDS-PAGE. RESULTS: DQD inhibited the platelet aggregation induced by thrombin (500 U/L), when DQD concentrations were 100, 200, and 400 mumol/L, the inhibition rates were 77%, 86%, and 82% respectively. DQD inhibited Ca2+ influx in platelets induced by thrombin (500 U/L) in the presence of extracellular Ca2+ 1 mmol/L in a concentration-dependent manner (10-80 mumol/L); DQD also had inhibitory effect on intracellular Ca2+ mobilization in the absence of extracellular Ca2+. DQD (10-160 mumol/L) inhibited the cytosolic Ca2+/PL dependent PKC from platelets in a concentration-dependent manner, but had no effect on membrane PKC. DQD (20-200 mumol/L) inhibited the actin polymerization induced by thrombin (500 U/L) in platelets in a concentration-dependent manner. CONCLUSION: DQD inhibited pig platelet aggregation induced by thrombin and its molecular mechanism was due to its inhibition of Ca2+ influx, intracellular Ca2+ mobilization, Ca2+/PL dependent PKC activity, and actin polymerization. PMID- 11501185 TI - Human hepatoma cell telomerase activity inhibition and cell cycle modulation by its RNA component antisense oligodeoxyribonucleotides. AB - AIM: To investigate the effects of human telomerase RNA component antisense oligodeoxyribonucleotides on telomerase activity of human hepatoma cells and their effects on cell cycle distribution. METHODS: Modified telomeric repeat amplification protocol was used to detect telomerase activity. Cell cycle was analyzed by flow cytometer. RESULTS: Telomerase activity was detected in all of four human hepatoma cell lines but absent in normal liver cells. Antisense oligomers to human telomerase RNA component (hTR) inhibited telomerase activity of BEL-7404 human hepatoma cells markedly in vitro. After in vitro treatment with antisense oligomers for 96 h, cell cycle of BEL-7404 human hepatoma cells was mainly arrested at G2/M phase. CONCLUSION: Antisense oligomers to hTR inhibited telomerase activity of BEL-7404 human hepatoma cells in vitro and resulted in cell cycle arrest at G2/M phase. PMID- 11501186 TI - In vitro proguanil activation to cycloguanil is mediated by CYP2C19 and CYP3A4 in adult Chinese liver microsomes. AB - AIM: To identify the cytochrome P450 isoforms involved in proguanil (PG) activation to cycloguanil (CG) in Chinese liver microsomes. METHODS: The kinetics of the CG formation from PG was determined in the liver microsomes of 6 Chinese subjects. Selective chemical inhibitors to various cytochrome P450 isoforms were employed to conduct inhibition experiments. The relationship between the CG formation and S-mephenytoin 4'-hydroxylation was analyzed. RESULTS: The kinetic behaviors of CG formation were described well by a single-enzyme Michaelis-Menten equation in five livers. The apparent Km and Vmax were (82 +/- 47) mumol.L-1 and (8 +/- 6) pmol.min-1.mg-1 protein, respectively. However, the remaining one displayed a two-enzyme kinetic behavior. Inhibition experiments showed that troleandomycin (100 mumol.L-1) and diethyldithiocarbamate (100 mumol.L-1), as potent CYP3A4 and CYP2E1 inhibitors, respectively, reduced the formation rate of CG by 81.1% and 47.23%, while quinidine (10 mumol.L-1), furafylline (20 mumol.L 1), and sulfaphenazole (10 mumol.L-1), which were inhibitors towards CYP2D6, 1A2 and 2C9/10, respectively, did not display significant inhibition. At a low PG concentration of 5 mumol.L-1, the CG formation correlated well with S-mephenytoin 4'-hydroxylation (r = 0.805, P < 0.05). Nevertheless, when a high substrate concentration (500 mumol.L-1) was used, the correlation coefficient decreased (r = 0.581, P < 0.05). CONCLUSION: The present study indicates that CYP3A4 and CYP2C19 are involved in PG activation to CG in adult Chinese liver microsomes. CYP2C19 played an important role in the clearance of PG at a substrate concentration close to in vivo therapeutic concentrations, while CYP3A4 gradually made a dominant contribution with the increase of PG concentration. PMID- 11501187 TI - Probing CYP2C19 and CYP3A4 activities in Chinese liver microsomes by quantification of 5-hydroxyomeprazole and omeprazole sulphone. AB - AIM: To develop an analytical method for simultaneous quantification of 5 hydroxyomeprazole (5-OH-OP) and omeprazole sulfone (OPS), and explore whether omeprazole (OP) is an appropriate phenotypic probe for CYP2C19 and CYP3A4 in Chinese liver microsomes. METHODS: OP metabolism in vitro was conducted in Chinese liver microsomes, and the major metabolites 5-OH-OP and OPS were determined using high pressure liquid chromatography (HPLC). Monoclonal antibodies anti-CYP2C8/9/19 and anti-CYP3A4 were employed to conduct inhibition experiments. The protein contents of CYP2C19 and CYP3A4 were quantified using Western blot analysis and densitometric scanning. RESULTS: 5-OH-OP and OPS gave a baseline resolution in the HPLC analysis. The detection limits for both compounds were 0.01 nmol and the recovery (98%-102%) had good precision with relative standard deviation of < 9.5%. Both anti-CYP2C8/9/19 and anti-CYP3A4 had a significant inhibitory effect (P < 0.05) on the 5-OH-OP formation in a substrate concentration-dependent manner, and anti-CYP3A4 alone could almost abolish the formation of OPS (> 87%). At a substrate concentration of 2 mumol/L OP, good correlations were found between OP 5-hydroxylation and S-mephenytoin 4' hydroxylation activities (r = 0.72, P < 0.01), OP 5-hydroxylation activities and CYP2C19 contents (r = 0.82, P < 0.01), and OP sulfoxidation activities and CYP3A4 contents (r = 0.78, P < 0.01) in Chinese liver microsomes. CONCLUSION: OP metabolism is mediated mainly by CYP2C19 and CYP3A4, and OP can be used to probe CYP2C19 and CYP3A4 activities in Chinese liver microsomes at appropriate substrate concentrations with the HPLC method presently developed. PMID- 11501188 TI - Regulation of telomerase activity in camptothecin-induced apoptosis of human leukemia HL-60 cells. AB - AIM: To explore the regulation of telomerase activity in camptothecin induced apoptosis of human leukemia HL-60 cells. METHODS: Apoptosis of HL-60 cells was induced by camptothecin (1 mg/L) for 2, 4, and 6 h. Apoptosis was determined by agarose electrophoresis and flow cytometry analysis. To assess telomerase activity, a PCR-based telomeric repeat amplification protocol assay (TRAP) was used. RT-PCR was performed to examine the mRNA levels of hTR (human telomerase RNA component), hEST2/hTERT (human telomerase reverse transcriptase), TLP1/TP1 (telomerase association protein 1), and bcl-2 (B cell leukemia/lymphoma 2 gene) in HL-60 cells before and after camptothecin treatment. RESULTS: Telomerase activity was decreased in a time-dependent manner during the camptothecin induced apoptosis of HL-60 cells. However, no difference in expression of each telomerase subunit was detected, while expression of bcl-2 was progressively down-regulated. CONCLUSION: These lines of evidences indicated that down-regulation of the telomerase activity in HL-60 cells was closely related to camptothecin-induced apoptosis, and the telomerase activity was not blocked at the transcriptional levels of the genes of the known ribonucleoprotein (RNP) complex components. We also found that bcl-2 did not regulate the mRNA expressions of telomerase subunits directly. PMID- 11501189 TI - Immunosuppressive effects of rubidatum in mice. AB - AIM: To study the effect of rubidatum (Rub) on immune function in normal mice. METHODS: Serum lysozyme concentration (SLC) was measured using micrococcus lysodiekticus as a substrate. Delayed type hypersensitivity (DTH) was determined by measuring the thickness of the right hind footpad 24 h after the injection of 1 x 10(8) washed sRBC (50 microL 10% sRBC). Serum hemolysin concentration was determined by OD measuring at A540 after the serum was treated with 2 mercaptoethanol. Phagocytic function of peripheral leukocyte (Leu) were determined by the incorporated radioactivity of [3H]TdR. The hemolytic activity of plaque forming cell (PFC) was determined by measuring the lymphocytemediated hemolysis of sheep red blood cell in vitro. T- and B-lymphocyte transformation (TLT and BLT) were induced by phytohemagglutinin (PHA) and lipopolysacharide (LPS) respectively and measured by the incorporation of [3H]TdR. RESULTS: Rub 125, 500, 2000 mg.kg-1.d-1 p.o. to BALB/c (or NIH) mice decreased the SLC; inhibited the phagocytosing functions of peripheral leukocytes; diminished the hemolytic activity of PFC; decreased the HC50; inhibited the DTH reaction; and showed inhibitory effects on TLT and BLT. CONCLUSION: Rub has immunosuppressive effects on immune system in mice by affecting M phi, T, and B lymphocyte, which suggests that Rub has inhibitory effects on both nonspecific and specific immune function. PMID- 11501190 TI - Evaluation of abnormal liver tests. AB - Serum liver tests are important but often problematic in evaluating patients with and without symptoms of hepatic disease. The common term "liver function tests" is misleading because most tests used in clinical practice measure hepatocellular damage not function. True liver function tests are those that measure synthesis of proteins made by the liver (albumin, clotting factors) or the liver's capacity to metabolize drugs. A commonly ordered panel of automated tests includes bilirubin, aminotransferases, alkaline phosphatase, and gamma-glutamyl transpeptidase. This article reviews patterns of elevated enzyme values encountered in liver diseases and their diagnostic limitations and provides an algorithm for evaluating abnormal liver test results. PMID- 11501191 TI - The role of liver biopsy in the evaluation of liver test abnormalities. AB - Percutaneous liver biopsy, first performed by Paul Ehrlich in 1883, remains an important diagnostic procedure for the management of hepatobiliary disorders. Modern biochemical, immunologic, and radiographic techniques have facilitated the diagnosis of liver disease but have not made biopsy obsolete. Indeed, examination of liver histology remains the "gold standard" for many diagnoses. This article reviews current indications for liver biopsy and the role biopsy plays in managing a variety of disorders. PMID- 11501192 TI - Treatment of hepatitis B. AB - Hepatitis B virus (HBV) is a major world health problem and a common cause of cirrhosis and hepatocellular carcinoma. The natural history of HBV varies with many factors, including age of acquisition. Persistent elevation of alanine aminotransferase (ALT) levels and presence of hepatitis B surface antigen for > 6 months after infection suggest chronic HBV. Presence of hepatitis B e antigen (HBeAg) and HBV DNA in serum indicate active disease. Treatment is indicated for chronic active HBV. The aim of treatment is to suppress viral replication and eliminate the virus. Endpoints of treatment are normalization of ALT levels and elimination of HBeAg and HBV DNA from the blood. Available treatments are interferon alfa and lamivudine. Interferon is effective in 25% to 40% of patients, but has serious side effects. Lamivudine is effective in a similar percentage of patients and has fewer side effects; however, it is associated with the emergence of viral mutations and drug-resistant strains. PMID- 11501193 TI - Treatment of hepatitis C. AB - With > 100 million people infected worldwide and > or = 4 million people infected in the United States, hepatitis C virus (HCV) is a major public health concern. In the United States, HCV is the most common cause of chronic liver disease. End stage liver disease from chronic HCV is the most common indication for liver transplantation. PMID- 11501194 TI - Fatty liver and nonalcoholic steatohepatitis. AB - Fatty liver is a relatively common incidental finding on imaging studies. Although generally a benign condition, fat in the liver can be troubling for clinicians because it can cause persistently elevated liver enzyme levels. The finding of fatty liver may also indicate the presence of nonalcoholic steatohepatitis (NASH). NASH is a histologic diagnosis applied to a constellation of liver biopsy findings that appear similar to alcoholic liver disease but are found in the absence of alcohol abuse. NASH is typically identified during the evaluation of elevated aminotransferase levels after exclusion of viral, metabolic, and other causes of liver disease. Obesity is a major risk factor; the role of diabetes is less certain, although evidence is accumulating that hyperinsulinism may play an important pathophysiologic role. About 15% to 40% of NASH patients develop hepatic fibrosis, a precursor to cirrhosis. Exactly how many patients with NASH progress to cirrhosis is unknown, but 1% to 2% of liver transplants are now performed because of a pretransplant diagnosis of NASH. Specific and effective treatments are needed but until the pathogenesis of this common liver disease is better understood, weight loss will remain the mainstay of treatment for obese patients. PMID- 11501195 TI - Mitochondrial toxicity. PMID- 11501196 TI - Evidence for mitochondrial toxicity: lactic acidosis as proof of concept. PMID- 11501197 TI - Clinical manifestations of mitochondrial toxicity. PMID- 11501198 TI - Mitochondrial dysfunction in HIV infection: an overview of pathogenesis. PMID- 11501199 TI - Clinical significance of treatment-induced lipid abnormalities and lipodystrophy. PMID- 11501200 TI - Cohort studies: what do they tell us? PMID- 11501201 TI - Effects of sex and race on lipodystrophy pathogenesis. PMID- 11501202 TI - Does antiretroviral-induced hyperlipidaemia constitute a cardiovascular risk? PMID- 11501203 TI - The use of lipid-lowering agents in HIV-infected patients. PMID- 11501204 TI - Tennessee medical schools: are we educating today's medical students for tomorrow's practices? PMID- 11501205 TI - Health policy and the law of unintended consequences. PMID- 11501206 TI - Loss prevention case of the month. Sympathy for family--no quality issues physician still loses. PMID- 11501207 TI - Medication errors: you can help reduce the numbers. PMID- 11501208 TI - Delivery of pharmaceutical care via the Internet. PMID- 11501209 TI - Access to prenatal, delivery, and newborn care among undocumented Hispanics in the Memphis area. AB - The purpose of this study was to examine the prenatal, delivery, and newborn care received by undocumented Hispanics in the Memphis area. The focus was to sketch a demographic profile of this population, identify barriers encountered in obtaining prenatal, delivery and newborn care, and present a five-year projection of the demand for prenatal care for this population. Participants included 97 undocumented Hispanic women, who either were pregnant or had a baby within the past 12 months, and health care providers. Most women came from Mexico (90.7%). They tended to be young, to have low educational attainment, to be homemakers, to have a high number of members in their households, to be financially supported by their spouses, and to live at or below the poverty line. The average length of time that they had been in the United States and Memphis were 42 and 21 months respectively. With regard to access to health care, undocumented Hispanics who migrated to the Memphis area lacked adequate prenatal and delivery care, and we estimated that the demand for prenatal care in this population will increase over 60% in the next five years. PMID- 11501210 TI - Cardiovascular disease: a clinical and public health problem. PMID- 11501211 TI - Interdisciplinary student fellowship program started by state government and the University of Tennessee. PMID- 11501212 TI - Quality of life--ageing and Down syndrome. AB - This article, based on pilot qualitative research, examines the quality of life of people with Down syndrome who are in the upper age bracket (45-70 years). Through use of a questionnaire, the current life experience and interests of a small group of individuals are noted, along with some of their perceptions concerning their past and present, including their views on the ageing process. Recommendations are made particularly in relation to the need to recognise the principles of variability, perception and choice, while providing support to encourage dignified and active lifestyles. PMID- 11501213 TI - Life be in it: lifestyle choices for active leisure. AB - For members of the community, participation in leisure, sports and recreation is an important lifestyle choice. Individuals with Down syndrome live in our community and they, too, are equally entitled to active lifestyle choices. Children, adolescents and adults with Down syndrome have a wide range of interests and, although reported trends indicate that their engagement in recreational activity is often sedentary and solitary in nature, other factors apart from the syndrome may account for this. Using a perception of difference perspective, this paper will examine certain aspects of their motor development, health and interactions with others which could be viewed as restrictive factors to their ability to participate in active leisure opportunities in the community. Program examples from Australia will be used to illustrate how a perception of difference which facilitates ability rather than disability across community based activities can enable a range of active leisure choices. PMID- 11501214 TI - Debunking the pathological model--the functions of an Internet discussion group. AB - In recent years there has been a profusion of resources about Down syndrome available on the World Wide Web and for increasing numbers of people the internet is the resource first contacted when new information is required on a topic. The present paper examined the function of a discussion group which is used primarily by parents of people with Down syndrome. Qualitative research methodology (content analysis) was used to analyse the daily discussions over a 5 month period in 1998 (February to July) and over a 2-week follow up period six months later (January 1999). The analysis revealed themes of celebration, seeing the child before seeing the handicap, hope and optimism, a sense of purpose in life and of being like a family within this group. PMID- 11501215 TI - Down syndrome in Israel. AB - The incidence of Down syndrome was studied in Jerusalem for the years 1964-1970 showing an overall incidence rate of 2.43 per 1,000 live births. A National Down Syndrome Register was established in 1978 and data on annual incidence and mortality rates from 1979-1997 is presented. The incidence in 1997 was 1.0 per 1,000 live births, but 2.32 per 1,000, when live births and terminated pregnancies are summed. Infant mortality has generally decreased in the past 20 years in Israel, and a decrease in infant mortality in Down syndrome has also been noted. This is due to better medical treatment and increased parental involvement in the care for infants with Down syndrome. PMID- 11501216 TI - Refractive errors and visual anomalies in Down syndrome. AB - A comparatively high incidence of ocular and orbital abnormalities has been reported in persons with Down syndrome. Eighty six children (50% male, 50% female) with Down syndrome in several institutions for individuals with learning difficulties (age range 5-18 years, mean 12.5) were examined for visual impairment in order to relate the ocular impairment to the level of learning difficulty. 6% had mild, 7% moderate, 45% severe and 42% profound learning difficulty. 9% of the children had no refractive errors. A significant (P < 0.01) positive correlation was found between progressive amounts of strabismus and ocular pathology with increasing amount of learning difficulty. On the other hand no correlation was found between refractive errors and the level of learning difficulty. Due to the significant number of ocular disorders found it is recommended that all children with Down syndrome should have an eye examination during the first six months of life and annually thereafter. PMID- 11501217 TI - Characterisation of the somatic evolution of Portuguese children with Trisomy 21- preliminary results. AB - We present preliminary results of a cross-sectional study which had the following objectives: 1--to develop percentile curves of weight, height and head circumference of Portuguese children with Trisomy 21 from 0 to 48 months of age; 2--a comparison of the growth of children with Trisomy 21 with a control population of their siblings, and 3--a comparison between the growth of Portuguese and American children with Trisomy 21 (based on the data of Cronk et al). We conclude that: 1--there is growth delay (weight, height, head circumference) in the Portuguese children with Trisomy 21, in all of the parameters evaluated and in all age groups; 2--Portuguese children with Trisomy 21 present values similar to those obtained by Cronk et al until 24 months of age; 3--from the age of 30 months onward Portuguese children with Trisomy 21 were heavier and taller than American children with Trisomy 21. This supports the usefulness of percentile curves specifically for Portuguese children with Trisomy 21. PMID- 11501218 TI - The relation between the psychological functioning of children with Down syndrome and their urine peptide levels and levels of serum antibodies to food proteins. AB - AIMS: To investigate the relation between psychological functioning of subjects with Down syndrome, and their levels of urine peptide and serum antibodies to food proteins. METHODS: 55 children with Down syndrome in a cross-sectional study. Psychological functioning was measured by the Stanford-Binet Intelligence Scale: Fourth Edition, McCarthy Scales of Children's Abilities and Fagan's computer based test of novelty preference. RESULTS: The participants, and their siblings, were found to have significantly increased total urine peptide levels. There were no significant correlations between peptide levels and psychological functioning. Significantly increased levels of IgG activity to gliadin and gluten, and IgA activity to gliadin, gluten and casein were found. There were significant negative correlations (Spearman r = -0.13 to -0.51) between psychological functioning, and IgG and IgA activity to gliadin and gluten. CONCLUSIONS: A significant relation between antibodies to gluten and psychological functioning was documented. The mechanism and potential causal link are still unknown. PMID- 11501219 TI - Acute coronary syndrome: are intervention and IIb/IIIa platelet inhibitors epiphenomena? PMID- 11501220 TI - Smoking cessation consensus. PMID- 11501221 TI - Hair as a biological specimen for therapeutic drug monitoring. AB - To study the potential of hair as a biological specimen in therapeutic drug monitoring (TDM), a review of the correlation between hair concentration, blood levels, dose and therapeutic effects of various drugs is presented. The results indicate there is a fair correlation between plasma and hair for several medicines. Moreover, hair samples have the advantage of providing information of more prolonged drug intake, thus allowing a better evaluation of patient compliance. Using segmental hair analysis, only a single sample is necessary. We conclude that hair has potential as a biological specimen in TDM--at least as far as compliance is concerned and possibly as a longer term record of drug concentrations--and that correlation between hair concentration, blood levels and clinical efficacy should be investigated for all drugs where TDM is indicated. PMID- 11501222 TI - Management of upper respiratory tract infection by family doctors. AB - The prescribing behaviour of family doctors in Hong Kong towards upper respiratory tract infections (URTIs) and the factors that might affect such behaviour were studied. All 1016 members of the Hong Kong College of Family Physicians were sent a postal questionnaire. A total of 730 family doctors completed the questionnaire, with an overall response rate of 71.8%. Doctors who were older, more senior, or in private practice were more likely to think antibiotics were useful for URTIs and to prescribe them. These doctors were also more likely to think their patients would expect antibiotics from them. Postgraduate vocational training in general practice/family medicine helped make family doctors think fewer patients would expect antibiotics from them but did not affect their perception of the usefulness of antibiotics for URTIs. Our results showed that doctors with certain characteristics were more likely to prescribe antibiotics for URTIs and these doctors may be targeted for continued medical education. PMID- 11501223 TI - The prevalence of pre-eclampsia and obstetric outcome in pregnancies of normotensive and hypertensive women attending a hospital specialist clinic. AB - To study the prevalence of pre-eclampsia (PE) and other obstetric outcomes (growth restriction and fetal mortality) in pregnancies of normotensive and hypertensive women attending an antenatal hypertension clinic, we studied a cohort of 372 pregnancies from 267 women. The prevalence of PE in the groups of pregnancies of normotensive and chronic hypertensive women was 11.9% (19/159 cases) and 16.0% (34/213 cases) respectively (chi 2 = 1.2, p = 0.27). There were no significant differences in respect of ethnicity, being primi- or multigravida and smoking status or age. Treatment with antihypertensive drugs during pregnancy did not decrease the prevalence of PE. In pregnancies with hypertensive complications (with or without PE) there was a trend towards higher rates of pre term delivery (< 37 weeks), caesarean section, small for gestational age babies, stillbirth and lower baby birth weight and ponderal index values. Pregnancies in women with uncomplicated hypertension had an increased risk for emergency caesarean section, pre-term delivery (< 37 weeks), birth weight < 2500 g and stillbirth (relative risks [with confidence intervals] 2.5 [1.9-3.2], 2.3 [1.8 2.9], 3.1 [2.5-3.7] and 5.5 [2.6-11.9] respectively) compared with the general hospital obstetric population. After classification according to the type of hypertensive syndrome, a progressively higher risk for fetal growth restriction and adverse perinatal outcome was shown in the hypertensive and pre-eclamptic groups. In chronic hypertension, this was irrespective of superimposed pre eclampsia or antihypertensive therapy. The high prevalence of PE in chronic hypertensive women (16.0%) was not statistically significant to that of normotensive women (11.9%), reflecting the referral selection of 'high risk' normotensive women to our clinic. PMID- 11501224 TI - The niacin skin flush test in schizophrenia: a replication study. AB - The aim of this study was to confirm a recent report that the non-invasive niacin skin flush test can be used to demonstrate impaired arachidonic acid-related signal transduction in schizophrenia. The response to topical aqueous methyl nicotinate solution was recorded at five-minute intervals over 20 minutes in 21 patients with schizophrenia, and in 20 age- and sex-matched normal individuals with no personal or family psychiatric history. The response was significantly lower in the patients with schizophrenia. At a concentration of 0.001 M, at the 15-minute timepoint, only two out of the 21 patients with schizophrenia showed a response, compared with 15 out of 20 of the controls (p < 0.00002), giving a sensitivity of the niacin skin test of 90% and a specificity of 75%. Our results are therefore consistent with the previous published report and suggest that this test may be useful clinically in the diagnosis of schizophrenia. PMID- 11501225 TI - The toxic oil syndrome: 20 years on. AB - With hindsight, it is easy to criticise the standards of food regulation of two decades ago. Nevertheless, when the Spanish toxic oil syndrome (TOS) appeared in 1981, there were many who asked why aniline was permitted as an official adulterant for imported French rape seed oil, and why such adulterated oils were often illegally refined in Spain and marketed without difficulty. This review brings up to date a comprehensive survey of the ensuing research published in 1995 and concentrates on recent significant findings. These include the identification of the refinery that produced the toxic oil, and the detection of oil contaminants with possible aetiological significance. Possible chemical links have been found between oil contaminants and those detected in L-tryptophan implicated in the eosinophilia-myalgia syndrome (EMS). There is good evidence that the initial pathogenetic mechanism is immunological. On metabolic evidence, it is suggested that not one, but a group of, toxic agents was responsible for TOS. PMID- 11501226 TI - Research into complementary/alternative medicine: an attempt to dispel the myths. AB - The high level of popularity of complementary/alternative medicine (CAM) renders rigorous research into its efficacy and safety an important task. A House of Lords select committee called for more CAM research and for the establishment of UK centres of excellence to carry it out. In view of the otherwise dire funding situation, this would be a necessary step. This initiative should be complemented by CAM professional bodies setting up their own research budgets. Any central body overseeing CAM research has to be seen to be rigorous and impartial. Strong opinions which often dominate CAM can be a hindrance to good research. Evidence based CAM should no longer remain a contradiction in terms. PMID- 11501227 TI - Mealtime glucose regulation in type 2 diabetes. AB - Glucose control remains the cornerstone of therapeutic strategies designed to reduce the risk of long-term complications in patients with type 2 diabetes. This article reviews a growing body of evidence suggesting that post-mealtime glucose spikes may exert an independent effect on diabetic complications, and discusses how a more effective strategy requires better focus on the regulation of mealtime glucose. New treatment options, specifically designed to restore a more physiological post-mealtime insulin profile, may provide better tools to ensure more comprehensive glycaemic control, which will allow for better prevention of micro- and macrovascular complications in type 2 diabetic patients. PMID- 11501228 TI - Smoking cessation: a consensus statement with special reference to primary care. AB - Nicotine addiction is a serious medical condition that needs to be treated like any other chronic disease. Primary care must play a key role in smoking cessation and offering help should be a routine part of primary care practice. As the most frequent opportunity for intervention lies within primary care, GPs should ensure that they raise the issue of stopping smoking at least annually with their smoking patients. When the smoker is ready to stop, the initial personal involvement of the GP is likely to increase the chance of a successful quit attempt. Follow-up may be with another healthcare professional. The support of healthcare professionals for the smoker who is motivated to quit, combined with appropriate pharmacotherapy, can substantially increase the chances of a successful quit attempt. Secondary care staff should also make every effort to help people to stop smoking and should communicate effectively with primary care- opportunities during hospitalisation are frequently missed. Specialist smoking cessation clinics have an essential role in providing more intensive specialist treatment and the expertise to partner and support the primary care effort. Smoking cessation is one of the most cost-effective healthcare interventions that can be made. PMID- 11501229 TI - Glatiramer acetate (Copaxone). AB - Glatiramer acetate (Copaxone) is a novel preparation of synthetic peptides composed of four amino acids. Laboratory studies have shown that it prevents, or modifies, experimental allergic encephalomyelitis, the animal model for multiple sclerosis (MS), in several mammalian species. Its mode of action has not been fully elucidated but it is known to induce suppresser T-cells, known to be deficient in MS, and competitively inhibits the effect of CNS myelin antigens, thought to be important in the pathogenesis of MS, through MHC blockade. Controlled clinical trials have shown it to improve the natural history of MS by reducing both the relapse rate and the resultant disability. GA shows similar efficacy to interferon-beta (IFN-beta) but with fewer systemic side-effects and appears to be better tolerated by patients. It has thus justified its place in the new era of disease-modifying treatments for MS. While the evidence suggests GA should be considered as first-line therapy in selected patients, its differing mechanism of action also gives patients and doctors the option of an alternative agent when the efficacy of IFN-beta is waning or side-effects predominate. PMID- 11501230 TI - Clinical rationale for rosuvastatin, a potent new HMG-CoA reductase inhibitor. AB - Coronary heart disease (CHD) is the leading cause of death worldwide, and effective treatment of hyperlipidaemia can prevent development of CHD and significantly reduce the risk for cardiovascular events and mortality in this disease. The advent of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) has revolutionised the treatment of hyperlipidaemia, but many patients receiving these drugs still do not achieve their therapeutic goals. Rosuvastatin (Crestor; formerly ZD4522) is a new, potent and long-lasting inhibitor of HMG-CoA reductase that is highly selective for hepatocytes. Its pharmacokinetics permit once-daily dosing, and a lack of oxidative hepatic metabolism results in a reduced potential for drug-drug interactions. Preliminary clinical results indicate that it produces rapid dose-related reductions in total cholesterol, low-density lipoprotein cholesterol, triglycerides, and apolipoprotein B that may exceed those achieved with other currently available statins. Increases in high-density lipoprotein cholesterol have also been observed. Rosuvastatin is also well tolerated, with no evidence of either hepato- or myotoxicity. It is hoped that new agents such as rosuvastatin may help to reduce the high global morbidity, mortality and associated costs of CHD and related vascular disorders. PMID- 11501231 TI - Major arterial occlusion by embolisation of an occult thrombus following thrombolytic therapy for myocardial infarction. AB - We describe a case in which thrombolytic therapy for acute myocardial infarction resulted in a major arterial embolism from an existing aortic mural thrombus. Clinicians should be alert to this rare complication of thrombolytics, because timely diagnosis and treatment may prevent serious morbidity and mortality. PMID- 11501232 TI - Does low tri-iodothyronine independently predict mortality in elderly hospitalised patients? AB - A fall in serum tri-iodothyronine (T3) is the earliest abnormality in thyroid hormonal function tests in non-thyroidal illnesses. Our study shows an association of low serum T3 with patient mortality in elderly hospitalised patients. PMID- 11501233 TI - Does balance control deficit account for walking difficulty in Parkinson's disease? AB - Patients with Parkinson's disease (PD) walk slowly, in part to compensate for their balance control deficit. We tested the effect of balance support to determine if walking performance in PD patients would improve. The sample consisted of unmedicated older adults with idiopathic Parkinson's disease who had poor balance control but no stooped posture, arthritis or muscle weakness. There was no difference in walking speed between unsupported and supported walking. The speeds were between those reported for disease-free older adults and older adults with muscle weakness and a history of falling. PD patients' walking difficulties, even while using a balance aid, may be partly explained by their set-changing problems. They frequently hold the cane off the ground when walking, suggesting their set-changing difficulty may be severe enough that using it aggravates their walking difficulty. Treatment of walking difficulty in PD patients should consider interventions other than those dealing only with balance control. PMID- 11501234 TI - Facial paralysis with an inflammatory parotid mass. AB - Facial paralysis in association with a parotid mass is usually associated with a diagnosis of malignancy. Benign parotid neoplasms and inflammatory processes resulting in facial paralysis are extremely rare. This report describes such a case and highlights some of the difficulties surrounding the diagnosis and management of these cases. PMID- 11501235 TI - Collecting duct carcinoma of the kidney associated with the sickle cell trait. AB - Collecting duct carcinoma of the kidney is a rare neoplasm that arises from the medullary collecting ducts. It has an aggressive clinical course and patients usually present with metastatic disease. Treatment approaches apart from surgery have been unrewarding. Renal medullary carcinoma is also an aggressive tumour which occurs predominantly in young, black, male patients with sickle cell trait. We describe a case of collecting duct carcinoma in a 61-year-old black female who exhibited the sickle cell trait. Histopathology revealed a high-grade tubular carcinoma with marked desmoplasia, adjacent dysplastic collecting ducts and positive immunohistochemical staining for high and low molecular weight cytokeratins and epithelial membrane antigen. The association of collecting duct carcinoma with sickle cell trait has not been previously reported. This case supports the postulate that collecting duct carcinoma and renal medullary carcinoma may be part of the same spectrum of renal malignancies. PMID- 11501236 TI - Congenital anosmia. AB - We present two cases demonstrating congenital anosmia. In both cases, magnetic resonance imaging (MRI) has served to highlight an interesting clinical oddity. To date, there has only been one study of the use of MRI in the assessment of patients with congenital anosmia who do not have Kallmann's syndrome. PMID- 11501237 TI - Word deafness presenting as a sudden hearing loss. AB - Sudden hearing loss is a condition frequently seen in ENT departments and is usually due to otological pathology. We present a patient who described symptoms of a sudden inability to hear but who, on further questioning and investigations, had 'word deafness' due to bilateral temporoparietal infarcts. We discuss the clinical syndromes associated with these infarcts and the specific management of word deafness. PMID- 11501238 TI - Superficial radial neuropathy following venepuncture. AB - A 42-year-old female suffered excruciating pain and paraesthesia on venepuncture of the cephalic vein in her left wrist. The left superficial radial nerve was injured. A flexed wrist during venepuncture renders the superficial radial nerve immobile and vulnerable to being punctured by the needle. To reduce the risk of nerve injury during venepuncture, the phlebotomist should choose a large and visible vein and insert the needle at a 5-15 degrees angle with the skin. The wrist should be selected only if the veins in the antecubital area are deemed unsuitable. The feeling of an electric shock along the distribution of the nerve, or rupture of the vein during venepuncture, should alert the phlebotomist to the possibility of nerve injury and the procedure should be stopped immediately. PMID- 11501240 TI - Confessions of a continuing education junkie: or, how I survived an excruciatingly boring dental lecture. PMID- 11501239 TI - New York State--dental establishments--keep getting larger. AB - Bureau of the Census data, through the end of the 1990s, indicate a continuing increase in the size of dental establishments. A review at the national, state and county levels document these developments for New York State. PMID- 11501242 TI - Voodoo Barbie and the dental office. PMID- 11501241 TI - Hemostatic technique. Using a splint in oral bleeding. AB - Controlling hemorrhage from dental treatment in bleeding disorder patients is one of the most serious procedures encountered by the dentist. Local hemostatic techniques combined with replacement therapy are the usual management. A polyurethane stent as an adjunct therapy is beneficial in controlling hemorrhage. The hemostatic stent provides pressure at the surgical site and adequate protection. PMID- 11501243 TI - When the insurance company demands a refund. PMID- 11501244 TI - Don't just get angry.... PMID- 11501245 TI - ADA responds to amalgam litigation. PMID- 11501246 TI - A very expensive vacation. Dentist pays ultimate price for failing to make adequate arrangements for covering his office while away. PMID- 11501247 TI - Preventing conduct problems, promoting social competence: a parent and teacher training partnership in head start. AB - Studied the effectiveness of parent and teacher training as a selective prevention program for 272 Head Start mothers and their 4-year-old children and 61 Head Start teachers. Fourteen Head Start centers (34 classrooms) were randomly assigned to (a) an experimental condition in which parents, teachers, and family service workers participated in the prevention program (Incredible Years) or (b) a control condition consisting of the regular Head Start program. Assessments included teacher and parent reports of child behavior and independent observations at home and at school. Construct scores combining observational and report data were calculated for negative and positive parenting style, parent teacher bonding, child conduct problems at home and at school, and teacher classroom management style. Following the 12-session weekly program, experimental mothers had significantly lower negative parenting and significantly higher positive parenting scores than control mothers. Parent-teacher bonding was significantly higher for experimental than for control mothers. Experimental children showed significantly fewer conduct problems at school than control children. Children of mothers who attended 6 or more intervention sessions showed significantly fewer conduct problems at home than control children. Children who were the "highest risk" at baseline (high rates of noncompliant and aggressive behavior) showed more clinically significant reductions in these behaviors than high-risk control children. After training, experimental teachers showed significantly better classroom management skills than control teachers. One year later the experimental effects were maintained for parents who attended more than 6 groups. The clinically significant reductions in behavior problems for the highest risk experimental children were also maintained. Implications of this prevention program as a strategy for reducing risk factors leading to delinquency by promoting social competence, school readiness, and reducing conduct problems are discussed. PMID- 11501248 TI - The efficacy of a universal school-based program to prevent adolescent depression. AB - Evaluated whether a universal school-based program, designed to prevent depression in adolescents, could be effectively implemented within the constraints of the school environment. Participants were 260 Year 9 secondary school students. Students completed measures of depressive symptoms and hopelessness and were then assigned to 1 of 3 groups: (a) Resourceful Adolescent Program-Adolescents (RAP-A), an 11-session school-based resilience building program, as part of the school curriculum; (b) Resourceful Adolescent Program Family (RAP-F), the same program as in RAP-A, but in which each student's parents were also invited to participate in a 3-session parent program; and (c) Adolescent Watch, a comparison group in which adolescents simply completed the measures. The program was implemented with a high recruitment (88%), low attrition rate (5.8%), and satisfactory adherence to program protocol. Adolescents in either of the RAP programs reported significantly lower levels of depressive symptomatology and hopelessness at post-intervention and 10-month follow-up, compared with those in the comparison group. Adolescents also reported high satisfaction with the program. The study provides evidence for the efficacy of a school-based universal program designed to prevent depression in adolescence. PMID- 11501249 TI - The prospective relation between dimensions of anxiety and the initiation of adolescent alcohol use. AB - Examined the relation between early anxiety symptomatology (generalized and separation) and initiation of alcohol use 4 years later in an epidemiological sample of 936 children (45% girls), assessed at ages 9, 11, and 13, while controlling for the effects of depression. Although earlier overall anxiety symptomatology was unrelated to later onset of drinking, children with early symptoms of generalized anxiety were found to be at increased risk for initiation of alcohol use, whereas children with early symptoms of separation anxiety were at decreased risk. The magnitude of these relations was equally strong for boys and girls. In addition, early depressive symptomatology was associated with increased risk for initiation of alcohol use in adolescence. Results indicate that it is important to consider specific dimensions of anxiety symptomatology when attempting to identify those individuals at risk for early initiation of alcohol use. PMID- 11501250 TI - Effects of cumulative prenatal substance exposure and environmental risks on children's developmental trajectories. AB - Examined the effects of cumulative prenatal substance exposure and cumulative environmental risk on the developmental trajectories of 278 infants, toddlers, and preschool children. Results indicated that both cumulative risk indexes were significantly correlated. Results also indicated that both indexes were related to decrements in developmental trajectories (intercept and slope) from 3 to 57 months of age. Both prenatal exposure and environmental risk added unique variance to the prediction of developmental level and rate of growth when entered after covariates (i.e., birth weight and sex). However, across a number of models with and without covariates, environmental risk accounted for more variance in developmental trajectories than did prenatal exposure. Implications are discussed. PMID- 11501251 TI - Gender and weight concerns in early and middle adolescence: links with well-being and family characteristics. AB - Studied sex and developmental differences in weight concerns in early and middle adolescence and links between concerns and adolescent well-being and family experiences. Participants were mothers, fathers, and older and younger siblings (Ms = 15 and 12.5 years, respectively) from 197, Caucasian, working-middle class, 2-parent families. Parents rated their gender role attitudes and adolescents rated their weight concerns, well-being, gender role orientations, and physical development. Girls reported more concerns than boys; body mass index (BMI) correlated with weight concerns for all youth. Controlling for BMI and pubertal status, weight concerns were linked to older girls' well-being; with physical characteristics controlled, mothers' gender attitudes explained older girls' weight concerns, and siblings' weight concerns explained those of older and younger girls and boys. PMID- 11501252 TI - Parental maltreatment and emotion dysregulation as risk factors for bullying and victimization in middle childhood. AB - Examined whether children who were maltreated by caregivers were more likely to bully others and to be at risk for victimization by peers. An additional focus was to investigate emotion's role in bullying and victimization among children at risk. Participants were 169 maltreated and 98 nonmaltreated boys and girls attending a summer day camp for inner-city children. As predicted, maltreated children were more likely than nonmaltreated children to bully other children. Bullying was especially prevalent among abused children who experienced maltreating acts of commission (physical or sexual abuse). Maltreatment also placed children at risk for victimization by peers. Gender did not moderate these findings, in that maltreated boys and girls appeared to be at similar risk for bullying and victimization. As expected, both bullies and victims evidenced problems with emotion regulation. Furthermore, logistic regression analyses suggested that emotion dysregulation made a unique contribution toward differentiating bullies and victims from children who did not evidence bully victim problems. In addition, maltreatment's effects on children's risk for bullying and victimization were mediated by emotion dysregulation. PMID- 11501253 TI - Social inhibition and overfriendliness: two-year follow-up and observational validation. AB - Studied socioemotional functioning in 3 groups (n = 15) of high-inhibited, low inhibited-overfriendly, and low-inhibited-low-overfriendly children at age 9, who were identified by parental ratings in a sample (N = 392) at age 7. Ratings of social inhibition and overfriendliness were stable over the 2-year period from age 7 to age 9. Validity of rated social inhibition and overfriendliness is established with behavioral observations, both predictively and concurrently. The groups are contrasted along the dimensions of social inhibition and overfriendliness with respect to socioemotional distress, social problem solving, empathic ability, and the disinhibitory phenomena of thrill seeking and hyperactivity. The high-inhibited group shows heightened socioemotional distress. The overfriendly group shows heightened levels of thrill seeking and hyperactivity. Social problem solving and empathy are less well developed in the inhibited and in the overfriendly group as contrasted to the low-inhibited-low overfriendly group. The results are interpreted in terms of risks for socioemotional problems at both ends of the dimension of social inhibition. PMID- 11501254 TI - The child PTSD Symptom Scale: a preliminary examination of its psychometric properties. AB - Reports on the development and preliminary validation of the Child PTSD Symptom Scale (CPSS) for children and adolescents. The CPSS is a new instrument that was developed to assess the severity of Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) posttraumatic stress disorder symptoms in children exposed to trauma. The CPSS was administered to 75 school-age children approximately 2 years after the 1994 Northridge, California, earthquake. The psychometric properties of the CPSS show high internal consistency and test-retest reliability for both the total score and the three subscales. Convergent validity with the Child Post-Traumatic Stress Disorder Reaction Index (CPTSD-RI) was established. As expected, the correlations of the CPSS with depression and anxiety measures were lower than those with the CPTSD RI, providing some support for discriminant validity of the CPSS. These results suggest that the CPSS is a useful tool for the assessment of posttraumatic stress disorder (PTSD) severity and for the screening of PTSD diagnosis among traumatized children. PMID- 11501255 TI - How much does physical appearance say about the psychological adjustment of competent and dysfunctional children? AB - Presents a study in which three sets of photographs of socially competent, aggressive, and anxious preschoolers were rated by college students (n = 150 raters per set), blind to the children's group membership. This was done to assess the extent to which adults are able to make valid and reliable evaluations of children's psychological adjustment on the basis of physical appearance alone. Sets 1 and 2 were photographs of different children taken under the same conditions and providing both facial and nonfacial cues. Sets 2 and 3 were of the same children taken under conditions that varied as to the amount of nonfacial cues they provided. Results showed that (a) socially competent children were judged to be better adjusted than their dysfunctional peers (i.e., more competent, less aggressive, less anxious, and less likely to have emotional or behavioral problems); (b) within the dysfunctional group, aggressive and anxious children were distinguished in ways that correspond closely to what is known about them from behavioral and clinical research; (c) irrespective of group membership, girls and boys were generally distinguished in ways that reflect normative beliefs about gender differences from social and developmental research; (d) group differences in ratings of psychological adjustment were generally comparable across photograph sets and could not be accounted for by differences in the children's perceived physical attractiveness; and (e) raters reported that they relied mainly on the children's expression, eyes, and posture to make their judgments of adjustment. These results replicate and extend earlier findings based on 1 of the 3 photograph sets (Serketich & Dumas, 1997). They suggest that when first impressions matter, competent children are at an advantage and their dysfunctional peers at a disadvantage even before their actual behavior comes to confirm or to invalidate these impressions. Theoretical and clinical implications of the findings are discussed. PMID- 11501256 TI - Assessing ADHD across settings: contributions of behavioral assessment to categorical decision making. AB - Adapted methods of behavioral assessment to assess home and school functioning in a way that maps directly to the Diagnostic and Statistical Manual of Mental Disorders (4th ed., [DSM-IV]; American Psychiatric Association, 1994). The study was conducted in a school-based sample with 5- to 12-year-old children referred to a school intervention team. A multigate set of procedures was used to assign children to one of 3 groups: attention deficit hyperactivity disorder (ADHD), inattentive group; ADHD, combined group; and a non-ADHD control group. The ADHD Rating Scale-IV was used to assess parent and teacher ratings of ADHD symptoms as delineated in DSM-IV. The findings suggest that the use of a fixed cutoff point (i.e., 6 or more symptoms), which is employed in the DSM-IV, is often not the best strategy for making diagnostic decisions. The optimal approach depends on whether diagnostic information is being provided by the parent or teacher and whether the purpose of assessment is to conduct a screening or a diagnostic evaluation. Also, the results indicate that a strategy that aggregates symptoms in the order in which they are accurate in predicting a diagnosis of ADHD is a more effective strategy than the approach used in DSM-IV, which aggregates any combination of a specific number of items. Implications for using methods of behavioral assessment to make diagnostic decisions using DSM-IV criteria are discussed. PMID- 11501257 TI - The effect of child characteristics on teachers' acceptability of classroom-based behavioral strategies and psychostimulant medication for the treatment of ADHD. AB - Studied the effect of student characteristics on teachers' ratings of treatment acceptability for attention deficit hyperactivity disorder (ADHD). Participants (N = 159) included experienced elementary school teachers who read 1 of 6 vignettes describing a child with symptoms representative of ADHD. Vignettes varied by sex and symptom-subtype classification. However, the number and specific type of symptoms described in the vignettes were consistent across all conditions. Next, teachers read a description of a daily report card (DRC), response cost technique, classroom lottery, and medication and rated their levels of agreement to the items of the Behavioral Intervention Rating Scale (BIRS). Teachers preferred the DRC to all other forms of treatment. However, there was a significant interaction between the type of treatment and sex of the student on the 3 factors (Treatment Acceptability, Treatment Effectiveness, and Timeliness) of the BIRS. PMID- 11501258 TI - Examining outcome variability: correlates of treatment response in a child and adolescent anxiety clinic. AB - Examined correlates of treatment response in a clinic providing cognitive behavioral therapy for children with anxiety disorders. Youth (ages 7 to 15) with a primary Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev., or 4th ed.; American Psychiatric Association, 1987, 1994) anxiety-disorder diagnosis (overanxious disorder, generalized anxiety disorder, separation anxiety disorder, social phobia, or avoidant disorder) participated. After completing a full course of treatment and posttreatment (n = 135) and 1-year follow-up (n = 107) assessments, participants were classified into 1 of 2 groups--poor treatment response and good treatment response--using parent diagnostic reports. Discriminant function analyses indicated that higher levels of maternal- and teacher-reported child-internalizing psychopathology at pretreatment, higher levels of maternal self-reported depressive symptoms, and older-child age were all associated with less favorable treatment response. Other factors, such as child ethnicity, child sex, family income, family composition (i.e., dual parent vs. single parent), child-reported symptomatology, and maternal-reported level of child-externalizing behavior problems did not predict treatment response. Both practical and conceptual implications of the findings are discussed. PMID- 11501259 TI - [Evolution of isolation and identification techniques of waterborne viruses]. AB - The paralytic potential of the poliovirus was recognized as early as the 14th century B.C. as illustrated in Egyptian art. But it is only after the four last decades that methods for their concentration from water and their identification were performed. Among several of them the adsorption-elution method was retained. Nevertheless two important barriers had to be ran-over. The first one was the concentration-elution steps on different materials which had to be improved. The second one was the typing method which had to move from particle by particle identification to entire viral population. Despite of these advances only a few cytopathogenic serotypes were found. The reverse transcriptase-polymerase chain reaction with its far more wide spectrum allows the fast and direct identification of viral nucleic acids (or their fragments) of almost all viruses, cytopathogenic or not. With this method elevated amounts of drinking water samples were found positive for several non cytopathogenic viruses. The sanitary significance of these results has still to be proved. PMID- 11501260 TI - [Inducible nitric oxide synthase expression and nitric oxide production by monocytes in systemic sclerosis]. AB - We investigated nitric oxide (NO) production and inducible NO synthase (iNOS) expression by cultured peripheral blood mononuclear cells (PBMC) in systemic sclerosis (SSc). Eighteen patients with SSc were compared to two control groups: 16 rheumatoid arthritis patients (RA) and 23 mechanical sciatica patients. The sum of nitrites and nitrates was determined by fluorimetry in sera and spectrophotometry in supernatants. Inducible iNOS was detected in cultured PBMC by immunofluorescence, immunoblot and flow cytometry with or without IL-1 beta + TNF alpha, IL-4 or IFN gamma from day 1 to day 5. NO metabolite concentrations in the plasma were lower in SSc (34.3 mumol/l +/- 2.63 SEM) than in RA (48.3 mumol/l +/- 2.2; p < 0.02) and sciatica (43.3 mumol/l +/- 5.24; p < 0.03) patients. iNOS was detected in cultured monocytes in the 3 groups but induction occurred on day 1 in RA, day 2 in sciatica and only on day 3 in SSc, whatever the stimulus. The concentrations of NO metabolites are decreased in SSc patients and the induction of iNOS in PBMC is delayed. Low levels of NO, a vasodilator, may be involved in vasospasm, which is critical in SSc. This may suggest therapeutic implications. PMID- 11501261 TI - [Prosopagnosia]. AB - True prosopagnosy preserves the perception of human faces. However, the latter faces cannot be identified even by patients who have kept an intact implicit recognition. After a thorough examination of the visual recognition disorder, other kinds of information may be involved like, on one hand, those of places, landscapes and building constructions or, on the other hand, those of animals. We observed that a transitorily prosopagnosic farmer was definitively unable to recognize his cows. Arguments for a right hemisphere dominance are discussed. PMID- 11501262 TI - [Impaired recognition of faces: implicit recognition, feeling of familiarity, role of each hemisphere]. AB - We report three observations of patients who suffered from impaired face recognition following cerebral lesions. Two had classical prosopagnosia, resulting from bilateral in one case and right unilateral occipito-temporal in the other. They could not differentiate famous face from unknown ones, and did not feel any familiarity. The third patient has a normal feeling of knowing, could distinguish between familiar and unfamiliar faces, but was unable to evoke any biographical information about the personalities. Prosopagnosic patients demonstrated, in an experimental condition of learning face-name pairs, implicit knowledge. We assume that these capacities were dependent of the activation of networks coding familiar faces in memory. Mental imagery of faces were normal in theses two cases. In addition, stimulation of mental imagery in the first patient improved implicit knowledge in forced choice tasks. These cases throws a light on the respective role of each hemisphere in face recognition. The right hemisphere is advantaged in perceptual analysis, and activates, from the perceived faces, mnestic systems which codes for previously encountered faces. It generates feeling of familiarity, probably by the way of specific systems which differs from, and completes, those allowing identification. The left hemisphere enable access to semantic-biographic knowledge in a conscious, verbal and explicit way. PMID- 11501263 TI - [Neuronal death: potential role of the nuclear enzyme, poly (ADP-ribose) polymerase]. AB - Poly(ADP-ribose) polymerase (PARP, EC 2.4.2.30) is known as a nuclear enzyme that is activated by DNA strand breaks to participate in DNA repair. It is also called poly(ADP-ribose) synthase (PARS) or poly(ADP-ribose) transferase (PADRT). In physiological conditions, PARP plays an important role in maintaining genomic stability. However, for several pathological situations, which include massive DNA injury (brain ischemia for example), excessive activation of PARP can deplete stores of nicotinamide adenine dinucleotide (NAD+), the PARP substrate, which, with the subsequent ATP depletion, leads to cell death. PARP activation appears to play a major role in neuronal death induced by cerebral ischemia, traumatic brain injury, Parkinson disease and other pathologies. PARP inhibitors (3 aminobenzamide and other compounds) and PARP gene deletion induced dramatic neuroprotection in experimental animals (rats, mice). Accordingly, these data suggest that PARP inhibitors could provide a novel therapeutic approach in a wide range of neurodegenerative disorders including cerebral ischemia and traumatic brain injury. PMID- 11501264 TI - [A human pluripotent stem cell in the blood of adults: towards a new cellular therapy for tissue repair]. AB - The presence in normal adult man of stem cells sharing the properties of embryonic stem cells opens new avenues for basic and therapeutic research. We describe a stem cell present in normal adult human blood, probably able to give rise to the "reserve" stem cells in charge of repair, present in different organs. These monocytoid circulating cells are able to transdifferentiate into several cell types. In normal man, they are almost quiescent and are strictly controlled by a special subpopulation of T lymphocytes. In diseases such as fibrosis and chondrosarcoma, these cells proliferate and the differentiated cells escape T lymphocyte control. As a consequence, these cells accumulate, giving rise in vitro to a tissue which evoke the lesions characterizing the disorder of the patient, showing spontaneously their pluripotentiality. Neural cell markers are present in this migrating cell, suggesting that pluripotent stem cells present in adult man may derive from the neural crest. These circulating cells could offer a source of stem cells for cellular and gene therapy provided the normal cells could be expanded, their transdifferentiation directed and the control by T lymphocytes maintained. PMID- 11501265 TI - [Reflections on some methodological errors in the evaluation of drugs. Ten years experience at the National Marketing Authorization Commission]. AB - A ten year experience at the national french marketing authorization committee has permitted to notice the most commonly methodological errors in the field of clinical research and, particularly, in the dossiers for drug approval: a frequent insufficient sample size resulting in lack of statistical power, an unsatisfactory optimal dosing research, a misuse of the so-called surrogate markers, an erroneous opinion about the meaning of the p value, an abusive claim for equivalence in non significant superiority trials, a misuse of unadjusted multiple comparisons and too much confidence in subgroup analysis results. PMID- 11501266 TI - [Possible role of the pineal gland in the pathogenesis of idiopathic scoliosis. Experimental and clinical studies]. AB - The unexpected finding in 1959 by Marie-Jeanne Thillard that pinealectomy in young chickens gives way to spinal deformities was confirmed by the authors. In another experiment they found that injected melatonine to the chick at adequate dose and at the same time as surgery, lessen or even totally prevents the occurrence of deformities. On the other hand, at too low dose or delayed after pinealectomy melatonine injection, may not prevent the deformity which will be persisting or even increasing. In a subsequent series of experiments on the rat, pinealectomy results in decreasing the plasmatic amount of melatonine as well as giving way to spinal deformities. The nature of these deformities observed here is dependent on the stature between of the animal. The normal quadrupede rat develops after pinealectomy a standard scoliosis. Inversely the scoliotic deformity occurs when the animal has been forced to a bipede condition, which may be achieved by removing its forelimbs when baby, then forcing it to stand and remain in erect posture by high enough feeding. Melatonine depressing and erect position are in two conditions, when associated, likely to give way to experimental scoliosis. In human, a low nycthemeral level of plasmatic melatonine is correlated with progressive scoliosis. The level of platelets calmoduline, when is normally modulated by melatonine, has been proved by Kindsfater to be increased in progressive scoliosis. Then raises the hypothesis that human idiopathic scoliosis may be due to an inherited disorder of neuro-transmitters from neuro-hormonal origin, associated with bipedal condition, where an horizontal localized neuro-muscular imbalance starts and produces the scoliotic deformity of the fibro-elastic and bony structures axial spinal pilar. PMID- 11501267 TI - [Humoral effectors and cellular targets of chronic arterial wall allorejection]. AB - Contrasting with acute rejection, chronic rejection of arterial allograft don't involve leukocyte passengers from the graft to the host. Chronic rejection involves the proteins of the major histocompatibility complexes. Experimental studies in rats showed that the process evolved in three stages: a first stage of histo-incompatibility recognition mediated by the graft endothelium, a second stage of immune antibody-dependent injury of allogenic smooth muscle cells of the media associated with inflammatory infiltration of the adventitia, lastly a third stage of scarring process including intimal proliferation and adventitial fibrosis. PMID- 11501269 TI - Pediatric care. PMID- 11501268 TI - Wipeout. AB - When prehospital providers transported this patient to the trauma center, they felt a bit awkward, to say the least. The patient appeared to be intoxicated, and had fallen from approximately three feet. Nevertheless, upon EMS' arrival, the patient was responding only to painful stimuli and was not moving his lower extremities. This prompted EMS to activate the trauma system and treat the patient accordingly: He was immobilized on a long backboard. During transport, however, the patient became responsive to verbal stimuli and began moving his lower extremities. When he was transferred to the ED staff, he appeared to be doing fine neurologically, except for the intoxication. The EMS crew felt a bit embarrassed for the activation of trauma services. Three hours later, however, the ED physician called the providers at their station to inform them that the patient had an unstable cervical spine fracture (see x-ray above), and their care was definitely appropriate. PMID- 11501270 TI - Speaking the same language: PSWN works to improve communications in EMS. PMID- 11501271 TI - Take it online moving classroom-based EMS training and education to the Internet. PMID- 11501272 TI - A is for airway: alternative airway devices. AB - There are many alternative airway devices available on today's market. The type of device that your department uses will depend on your medical control, as well as local protocols and regulations. These devices all require practice, refresher training and on-going skill maintenance. Endotracheal intubation remains the gold standard for airway control; however, proper usage of an alternative airway device can allow patients to be ventilated and oxygenated, even if they cannot be immediately intubated. PMID- 11501273 TI - You did what? Clinical errors in EMS. AB - It was late and you were exhausted. You ran a call that didn't go the way you would have liked. Errors occurred. What actions do you take when your partner is making mistakes? When do you step in? Do you step in? What if there's more to the errors than you realize? This article provides guidelines for EMS personnel to consider when dealing with a scenario that contains clinical errors. Although not an all-inclusive list, the examples and solutions may be beneficial to providers, including rookies and veterans. PMID- 11501274 TI - Top of the line. 10 new product innovations from EMS EXPO 2001. PMID- 11501275 TI - Before the call: how EMS outreach can help kids. PMID- 11501276 TI - International EMS. Hawaii EMS challenged by powerful surf. PMID- 11501277 TI - Sublethal effects of chronic exposure to an organochlorine compound on northern leopard frog (Rana pipiens) tadpoles. AB - Global contamination with organochlorine compounds (OCs) has posed developmental and reproductive problems in wildlife worldwide. However, little is known about the impact of OCs or other pollutants on amphibians, despite mounting concerns about amphibian population declines and developmental deformities in the wild. Wildlife populations may be affected critically by sublethal impacts of anthropogenic disturbances, yet little research has focused on such effects in amphibians. In the current study, northern leopard frog (Rana pipiens) tadpoles were chronically exposed to a polychlorinated biphenyl (PCB) congener, 77-TCB, and effects on behavior, morphology, competitive performance, and corticosterone content were determined. R. pipiens activity levels and feeding rates were decreased by 77-TCB exposure, but morphology of mouthparts and body proportions were unaffected. 77-TCB enhanced growth and altered competitive interactions between R. pipiens and wood frog (Rana sylvatica) tadpoles. R. pipiens tadpoles exposed to 77-TCB showed decreased whole-body corticosterone content compared to controls both before and after injection with adrenocorticotropic hormone (ACTH). All of the factors examined in the current study play critical roles in tadpole development, growth, survivorship, and eventual reproductive success, suggesting negative population-level consequences for amphibians in PCB-contaminated habitats. PMID- 11501278 TI - Co-production of microcystins and aeruginopeptins by natural cyanobacterial bloom. AB - The relationship between Microcystis composition and the production of microcystins and nontoxic peptides in bloom cells, which was regularly collected in Lake Suwa, Japan, in the summer season from 1991 to 1994, was investigated. In order to determine the structures of the nontoxic peptides, we collected large amounts of bloom materials from the same lake on July 23, 1991, and isolated three nontoxic peptides. They were named as aeruginopeptins 917S-A, -B, and -C, and their structures were mainly determined by a mass spectrometry/mass spectrometry (MS/MS) technique as 19-membered cyclic depsipeptides possessing the Ahp (3-amino-6-hydroxy-2-piperidone) moiety. An analysis of the microcystins and aeruginopeptins in the collected blood cells and their Microcystis composition suggested that the M. aeruginosa large cell size produces both microcystins and aeruginopeptins, and the production of both compounds is genetically closely related. PMID- 11501279 TI - Factors affecting biodegradation of 2-chlorophenol by Alcaligenes sp. in aerobic reactors. AB - The influence of variations in carbon source concentration, cell inocula, pH, presence of other substrates, and other organisms on the biodegradation of 2 chlorophenol (2-CP) was studied for Alcaligenes sp. isolated from natural sources. Assays of biodegradation were performed in batch and continuous-flow fluidized-bed aerobic reactors. Evaluation of biodegradation was performed by determining total phenols, chemical oxygen demand (COD), and 2-CP by ultraviolet (UV) spectrophotometry. Measurement of microbial growth was carried out by the plate count method. Bioassays of acute toxicity were performed to evaluate detoxification by using Daphnia magna. Results obtained show that under batch conditions with initial inocula of 10(6) cells/mL the strain grew exponentially with 100, 200, and 300 mg/L of 2-CP within 48 hr. A lag period was observed with low cell density inocula (10(5) cells/mL). The strain showed marked delay in the biodegradation of 2-CP at pH 5. Removal of target substrate from mixtures containing other carbon sources demonstrated the possibility of concurrent growth. Mineralization of 2-CP was assessed by gas chromatography carried out at the end of the batch assays and at the exit of the continuous-flow reactor. The presence of other organisms (bacteria, rotifers, ciliate, and algae) that developed in the fluidized-bed reactor did not affect the efficacy of the biodegradation of 2-CP. The removal of 2-CP in the two assayed systems was over 97% in all cases. Toxicity was not detected at the exit of the continuous reactor. PMID- 11501280 TI - Effects of the organophosphorus insecticide fenitrothion on growth in five freshwater species of phytoplankton. AB - The acute toxicity of the insecticide fenitrothion was measured using four freshwater algae (Chlorella saccharophila, Chlorella vulgaris, Scenedesmus acutus, and Scenedesmus subspicatus) and one cyanobacteria (Pseudanabaena galeata). Insecticide concentrations eliciting 50% growth reduction over 96 hr (EC50) ranged from 0.84 to 11.9 mg/L. Fenitrothion was more toxic than other pesticides studied with the same algal species such as chlorsulfuron, molinate, and pyridaphenthion. The transformation of effective concentrations of fenitrothion and other pesticides obtained from toxicity measurements into percent of the saturation level in water is used as a first evaluation of potential hazard to aquatic systems. The insecticides fenitrothion and pyridaphenthion were less hazardous than the herbicides atrazine, benthiocarb, cinosulfuron, chlorsulfuron, methyl-bensulfuron, and molinate. The two species of Chlorella and the cyanobacterium Pseudanabaena were more tolerant to fenitrothion than the two species of Scenedesmus. PMID- 11501281 TI - Detection of DNA damage in two cell lines from rainbow trout, RTG-2 and RTL-W1, using the comet assay. AB - Screening methods to indicate the genotoxic potential of individual chemicals or environmental mixtures rely mainly on short-term bacterial tests. Differences in the genotoxic response of prokaryotic and eukaryotic cells necessitate the development of nonbacterial screening assays. A promising approach for this purpose could be the comet (single-cell gel electrophoresis) assay performed with fish cells in vitro. In the present study, we evaluated the comet assay with two different fish cell lines from rainbow trout (Oncorhyhnchus mykiss), the fibroblast-like RTG-2 cell line established from gonad tissue, and the epitheloid RTL-W1 cell line established from liver tissue. The cells were exposed in vitro during 2 hr to the genotoxins, 4-nitroquinoline-1-oxide (NQO), and benzo(a)pyrene (BaP), as well as to environmental samples. The LOEC values for NQO were similar in both cell lines, whereas for BaP, the RTL-W1 cells were found to be more sensitive than the RTG-2 cells. The slopes of the concentration-response curves of the two test compounds differed between the two cell lines, with RTG-2 cells showing a steeper slope for NQO, and RTL-W1 cells showing a steeper slope for BaP. When exposed to environmental samples from a remediation site, the RTL-W1 cell line, but not the RTG-2 cell line, indicated a genotoxic potential of the samples. The differences in the genotoxic response pattern of the two cell lines could be only partly explained in relation to metabolic enzymes, cytochrome P4501A, glutathione-S-transferase, and xenobiotic reductase. The findings of this study demonstrate that the comet assay with fish cell lines is suitable as in vitro screening assay in environmental genotoxicity testing, but the choice of test cell line may be critical. PMID- 11501282 TI - Time-dependent accumulation of cyanobacterial hepatotoxins in flounders (Platichthys flesus) and mussels (Mytilus edulis) from the northern Baltic Sea. AB - There is only limited information about the accumulation of algal toxins in aquatic organisms in the Baltic Sea. In this study we measured total cyanobacterial hepatotoxin levels in blue mussel (Mytilus edulis) and flounderi (Platichthys flesus) tissues. Flounder were caught with gillnets from the western Gulf of Finland during July and August 1999. Blue mussels were collected from an enclosure at 3 m depth and from an artificial reef (wreck, 25-35 m depth) in the western Gulf of Finland between June and September 1999. Flounder liver and muscle samples and soft tissues of mussels were analyzed for the cyanobacterial hepatotoxins (nodularin, NODLN and/or microcystins, MCs) using an enzyme-linked immunosorbent assay (ELISA). Results showed a time-dependent accumulation of hepatotoxins in flounder and mussels. In flounder, the maximum concentration 399 +/- 5 (sd) ng NODLN or MC/g dry weight (dw) was found in the liver of specimens caught on 21 August 1999. No hepatotoxins were detected in muscle samples. The maximum concentration of 2150 ng +/- 60 (sd) ng hepatotoxin/g dw was found in the mussel soft tissues collected on 20 August 1999. Temporal NODLN or MC trends indicated depuration of cyanobacterial hepatotoxin from mussels at surface level and an increase in NODLN or MC concentrations in those from the sea bed. These studies showed that despite the low cyanobacteria cell numbers the cyanobacterial hepatotoxins can accumulate in flounder and mussels. This may allow the further transfer of cyanobacterial hepatotoxins in the food web. PMID- 11501283 TI - Degradation of the cyanobacterial hepatotoxin microcystin by a new bacterium isolated from a hypertrophic lake. AB - A bacterium capable of degrading microcystins-RR, -YR, and -LR was isolated from a hypertrophic lake. The bacterium, designated Y2 and classified phenotypically as a member of the genus Sphingomonas, was shown to be distinct phylogenetically from any established species of Sphingomonas on the basis of 16S rDNA sequencing. The bacterium was tentatively identified as Sphingomonas by manual chemotaxonomy, but 16S rRNA sequencing analysis suggests that it is in fact a new species or even a new genus. When the Y2 bacterium was added to microcystins present in culture medium, the microcystins were degraded thoroughly in 4 days. The highest degradation rates of microcystins-RR and -LR were 13 and 5.4 mg L-1 day-1, respectively. The degradation rates were strongly dependent on temperature and the maximum rate was at 30 degrees C. PMID- 11501284 TI - Effect of linear alkylbenzene sulfonate on population growth of Daphnia galeata: a life table evaluation. AB - The chronic effect of linear alkylbenzene sulfonate (LAS12) on a daphnid species (Daphnia galeata) was examined by the life table experiment. The estimated responses in the intrinsic rate of population growth r were analyzed with two alternate concentration-response functions, i.e., the power function and the quadratic function. Based on the best-fit power function model with biases corrected by the jackknife procedure, the population-level EC50, which is defined as the concentration of chemicals that reduces the population growth rate (the intrinsic rate of natural increase) by 50%, was estimated as 2.5 mg/L. The 48-h acute immobility test yielded EC50 of 4.6 mg/L. The population-level effect of LAS12 on this test species is considerably more sensitive than the acute lethal effects to neonates. PMID- 11501285 TI - Acute toxicity modeling of rainbow trout and silver sea bream exposed to waterborne metals. AB - Of three proposed acute toxicity models, the uptake-depuration (UD) model, the time-integrated concentration (TIC) model, and the concentration-time (CT) model are derived and verified with acute toxicity data to estimate the internal residues of waterborne metals in fish as a function of a few constants and variables. The main factors are the exposure time, the external exposure concentration, the bioconcentration factor (BCF), and the depuration rate constant (k2). The UD model is based on the concept of residue levels at the cell membrane well correlating with the whole-body concentrations, whereas the TIC and the CT models are based on the idea of irreversible inhibition of the enzyme acetylcholinesterase (AChE) governing the metal acute toxicity in that metals in the entire fish or in the aqueous phase can be described by the critical area under the time-concentration curve that is associated with a critical TIC of toxicant in the target tissue. A highly significant correlation (r2 > 0.9) was found between predictions and LC50(t) data for both the TIC and the CT models, indicating successfully describe 4- to 18-d LC50(t) data of arsenic (As), cobalt (Co), copper (Cu), and Co/Cu mixture in rainbow trout (Oncorhyuchus mykiss) and of Cu in fingerlings and subadults of silver sea bream (Sparus sarba). The time dependent lethal internal concentration at the site of action that causes 50% mortality is also predicted for a given compound and species. It concludes that the TIC and the CT models can be applied to regulate the acute toxicity and to estimate incipient LC50 values and internal residues of waterborne metals in fish. PMID- 11501286 TI - Effect of cadmium on reproduction of daphnids in a small aquatic microcosm. AB - Chlorella vulgaris colonies were grown as nutrient agar for one week, followed by addition of Daphnia magna in overlying water for a second week. A range of concentrations of cadmium were added to the Chlorella or to the Chlorella followed by higher cadmium concentrations in the overlying fluid. A dose dependent reduction in reproduction was observed with increased cadmium. When parent daphnids were first exposed to a lower cadmium concentration, and their offspring of preconditioned juvenile daphnids then exposed to a higher concentration, cadmium tolerance in new-born daphnids was observed. PMID- 11501287 TI - Marriage through abduction ('Telefa') in rural north west Ethiopia. AB - A community based cross sectional study was conducted in a rural district of North West Ethiopia between February and April 1997 to determine the magnitude of marriage through abduction ('Telefa') and identify problems associated with it. Randomly selected and currently married 1,168 women were interviewed. The prevalence of marriage through abduction was 6.2% (72/1168). All the abductions reported were only once in lifetime during the first marriage. The median age at first marriage of abducted women was 13 years with a range of 13 (Minimum = 7 and Maximum 20). About two third (66.7%) of abducted women had been married more than once in their life time. Following a multivariate analysis in a logistic regression model abducted women were likely to be victims of abortion [Adjusted OR (95% CI) = 1.71 (1.10-3.05)], marital instability [Adjusted OR (95% CI) = 1.87 (1.10-3.18)], rape [Adjusted OR (95% CI) = 7.77 (3.78-15.95)] and domestic violence [Adjusted OR (95% CI) = 1.69 (1.11-2.81)]. The recognition of the magnitude and the associated health problems of marriage through abduction (Telefa) is important. Appropriate strategies that address the health needs of abducted women must be designed. Enforcing the judiciary system to discourage this harmful practice and empowerment of young girls and rural women is needed. PMID- 11501288 TI - Otitis media seen in Yekatit 12 Hospital. AB - Two thousand three hundred and thirty four selected patients with ear problem were seen in one of the ENT out patients department of Yekatit 12 Hospital from September 1994 to August 1996 were prospectively studied. Most of the patients came from Addis Ababa. Major clinical presentations were ear pain, purulent ear discharge, uni or bilateral decreased hearing ability. Patients were carefully evaluated by physical examination, otoscopy examination of the ear, schuilers view of the mastoid bone, audiometry findings and pus culture results. Diagnosis of these patients revealed that 1,630 (69.8%) had otitis while the rest 704 (30.16%) had other ear problems like ear wax, tinnitus and otitis external. One hundred two hundred thirty two patients (52.8%) had chronic otitis media with purulent discharge and decreased hearing ability, while 245 patients (10.5%) had chronic otitis media with out purulent discharge but with decreased hearing ability. Ninety six patients (4.1%) had acute otitis media and 57 patients (2.4%) had chronic seromucinous otitis media with decreased hearing ability. The micro organisms identified include klebsiella spp. (28.97%), E. coli (10.7%), Citrobacter (3.6%), Acinetobacter (4.7%), S. aureus (3.57%), P. Valgaris (4.3%), S. Epidermidis (4.54%) and Dephtheroids (7.3%). Eighty nine patients with acute otitis media were completely cured and regained their hearing ability, the rest did not regain their hearing ability despite treatment. A combination of antibiotics and surgical treatment is recognized as the most effective intervention in this situation. The management of the chronic draining ear are: 1). To achieve a clean healed dry ear, 2) to obtain an air-containing middle ear space, 3) to rehabilitate hearing. The prevention of deafness and treatment of chronic draining ear disease especially in children is surgery which is not yet fully practiced in our centers. PMID- 11501290 TI - Variations of the hepatic and cystic arteries among Ethiopians. AB - The anatomy of the hepatic and cystic arteries were investigated in 110 postmortem and cadaveric subjects. The right hepatic artery took origin from the proper hepatic artery (66.3%), the common hepatic artery (18.2%), the superior mesenteric artery (8.2%) or the celiac trunk (7.3%). Ten cases of accessory right hepatic artery originating from the superior mesenteric artery (7 cases), gastroduodenal artery (2 cases) or the left hepatic artery (1 case) were observed. The origin of the left hepatic artery included the proper hepatic artery (71.8%), the common hepatic artery (16.4%), the celiac trunk (10.9%) and the splenic artery (0.9%). The 14 cases of accessory left hepatic arteries originated from the common hepatic artery (5 cases), right hepatic artery (3 cases), gastroduodenal artery (2 cases) or the celiac trunk (4 cases). An extrahepatic branch to the quadrate lobe of the liver, also known as the middle hepatic artery, was observed in 47.3% arising mainly from the right or left hepatic arteries (20% each), the superior mesenteric artery (2.7%) and from the gastroduodenal artery (4.6%). The cystic artery mainly arose from the right hepatic artery (75.5%) but also took origin from the middle hepatic artery (12.7%), gastroduodenal artery (7.3%) or the left hepatic artery (4.5%). When the cystic artery is to the left of the common hepatic artery at its origin (39.1%), it crossed from left to right anterior to the common hepatic duct (28.2%) or posterior to the duct (10.9%). Irrespective of its relationship with the common hepatic duct, the cystic artery passed in the triangle of Calot in 89 cases. There were 11 accessory cystic arteries arising from the right hepatic (6 cases), the middle hepatic (3 cases) or the left hepatic arteries (2 cases). Arterial variations seen in the present study were significantly higher (p < 0.05) than that reported in the literature. This was mainly due to the variations seen in origin of the right hepatic artery in the female subjects which was significantly higher (p < 0.05) than in the male subjects. The significance of this finding needs further investigation. PMID- 11501289 TI - Retinopathy in patients of Tikur Anbessa Hospital diabetic clinic. AB - A total of 302 diabetic patients were selected from regular attendants of the Tikur Anbessa Hospital (TAH) diabetic clinic to determine the prevalence of retinopathy from December 1994 to March 1995. The mean age was 41.4 +/- 14.4 years (range 14-85). There were 160 males (53%) and 142 females (47%). One hundred forty (46%) were type 1 and 162 (53.6%) were type 2. The mean duration of diabetes was 9.4 +/- 5.4 years and the mean Hemoglobin Alc (HbAlc) was 10.4 + 2.2%. On the day of the examination the mean fasting blood glucose (FBG) and random blood glucose (RBG) were 195.5 +/- 79.9 mg/dl and 273.1 +/- 114.5 mg/dl respectively. The mean serum total cholesterol, triglycerides, LDL, VLDL and GDL were 166.5 +/- 45.5 mg/dl, 129.9 +/- 92.4 mg/dl, 94.5 +/- 36.4 mg/dl, 24.4 +/- 15.1 mg/dl and 44.3 +/- 11.5 mg/dl respectively. The overall prevalence of retinopathy was 37.8% out of which 108 patients (36.1%) had background retinopathy and 5 patients (1.7%) had proliferative retinopathy. The retina could not be visualized in three patients because of dense cataract. Retinopathy correlated positively with age, duration of diabetes and blood pressure respectively, however no significant correlation was seen with mean total HgAlc and serum lipids. Prevalence of retinopathy was comparable in type 1 and type 2 (p > 0.05). The prevalence of retinopathy in our patients relative to the duration of diabetes mellitus is high. Therefore, improving facilities for the diagnosis and treatment of retinopathy is recommended. PMID- 11501291 TI - Case of tuberous sclerosis. AB - Tuberous Sclerosis (TSc) is a benign multi-system hamaertomatosis and is one of the neurocutaneous syndromes (2, 3, 7). The first case of tuberous sclerosis in a 27 years old female patient is reported from Ethiopia. The importance of meticulous evaluation of a patient as a whole to reach at the right diagnosis is stressed. The clinicopathological features of tuberous sclerosis are discussed with literature review. PMID- 11501292 TI - A concomitant occurrence of esophageal and cervical carcinoma in a sixty year old female patient from Bale Zone of Oromia Regional State, a case report with literature review. AB - Oesophageal carcinoma usually presents with difficulty of swallowing initially for solid food materials later even for liquid diets. Cervical carcinoma presents with pain and bleeding per vaginum during intercourse (sexual contact). We report a concomitant occurrence of squamous cell carcinoma of esophagus and cervix in a 60 years old female patient from Bale Zone of Oromia Regional State, who initially presented with vaginal bleeding of 2 years duration and later with difficulty of swallowing of 9 months duration. To our knowledge this is the first report of a concomitant occurrence of esophageal and cervical carcinoma in Ethiopia. PMID- 11501293 TI - Mental health services and epidemiology of mental health problems in Ethiopia. PMID- 11501295 TI - Prevalence and antibiotic susceptibility pattern of bacterial isolates from blood culture in Tikur Anbassa Hospital, Addis Ababa, Ethiopia. AB - Between Mid-1996 and Mid-1998, 238 bacteria strains isolated from blood culture of adult patients of Tikur Anbassa Hospital, Addis Ababa, Ethiopia, were retrospectively analyzed for their frequency of isolation and antibiotic susceptibility pattern. Coagulase negative Staphylococcus aureus (CNS) were isolated with the highest frequency 103 (43.3%), followed by Staphylococcus aureus 34(14.3%), Klebsiella spp. 23(9.7%), E. Coli 19(8.1%), Pseudomonas spp. 16(6.7%), Acinetobacter spp. 12(5%), Salmonella spp. 9(3.8%) and miscellaneous group 22(9.2%). The gram positive bacteria constituted 149(62.6%) of the total blood isolates. It is suggested that a proportion of both the gram positive and gram negative isolated represent contaminants at blood sampling. Rates of susceptibility for gram positive range from 12% to 76%, and for gram negatives range from 8% to 46%. In general, rates of susceptibilities to all antibiotics tested for gram negatives were very low as compared to gram positives. Among the gram positives, more than half of the isolates were sensitive to amoxicillin + clavulanic acid, ampicillin, carbenicillin, cephalothin, chloramphenicol, erythromycin and methicillin. Gram negative bacteria showed a high rate of resistance to many of the commonly prescribed antimicrobial drugs: amoxicillin + clavulanic acid (65%), ampicillin (87.5%), amoxicillin (91.7%), carbenicillin (75%), cephalothin (73.6%), chloramphenicol (65%), gentamicin (55.6%), kanamycin (54%), trimethoprim-sulphamethoxazole (64%) and tetracycline (61%). If generally considered, only gentamicin and kanamycin were relatively effective against gram negatives. Over 85% Salmonella spp were sensitive to chloramphenicol and trimethoprim-sulphamethoxazole. Compared to previous studies done in the same hospital, there is a higher rate of antibiotic resistance for most types of blood culture isolates particularly for gram negatives. The rational use of drugs should be practiced in order to minimize the spread of drug resistant bacteria. PMID- 11501296 TI - A typology approach to describing parents as communicators about sexuality. AB - Teenagers in Grades 8 and 10 and their parents completed a questionnaire examining the frequency of parental communications about sexuality and the communicative style when discussing sexuality and in general. Respondents also assessed parents' competence in communicating about sexual matters. For each set of respondents (teens reporting about mother, teens reporting about father, mothers' self-reports, fathers' self-reports), a cluster analysis yielded four clusters that were similar for each set. Relative to other parents, there was a group of parents that could be labelled as competent communicators and a group that could be labelled as problematic communicators about sexuality, with strong associations between cluster membership and score on the global measure of communicative competence. There were two intermediate categories that reflected more or less competence although the precise nature of these clusters differed as a function of informant group. Overall, fathers were rated as poorer communicators about sexuality than were mothers, at least by their teenage children. Consistent with other studies, mothers were more likely to be perceived as effective communicators by daughters and older teens. It appears that, independent of their level of competence, parents adjust their communication strategies according to the age and sex of their child, at least in the eyes of that child. Effective and problematic communicators among mothers were regarded as such by both sets of informants. This was not the case for fathers. We conclude that it is possible to classify parents usefully on the basis of perceived competence as communicators about sexuality. PMID- 11501297 TI - Sexual child abuse in a defined Swedish area 1993-97: a population-based survey. AB - Attempting to avoid some of the most common methodological problems involved in research on sexual child abuse, we collected data on crimes, perpetrators, and sanctions in all convicted cases of sexual child abuse in a defined population during a 5-year period. This approach provided amply documented and ascertained cases with precise definitions and descriptions of the crimes involved, no clinical referral bias, and minimal dependence on memory effects. The results are valid for the small proportion of cases that lead to conviction in the context of Swedish legislation. Structured data were collected from the court dossiers in all cases of sexual crimes against minors (less than 15 years of age) tried and sentenced at the courts in the Vastra Gotaland region of Sweden between 1993 and 1997. The total number of 496 sentences for sexual crimes during the study period included 203 cases of sexual child abuse (40.8%) with 283 victims and 196 perpetrators, all men. Girls were victims in 85% of the cases, boys in 12%, and boys as well as girls in 3%. Sexual penetration had occurred in 54.5% of cases and the total proportion of hands-on crimes was 83%. Most perpetrators, 72%, were well known to the child. The most severe offenses took place within the family. A wide range of acts were classified as sexual child abuse, but most common was sexual penetration of a female child by her biological father or a family friend. PMID- 11501298 TI - An evaluation of self-report measures of cognitive distortions and empathy among Australian sex offenders. AB - The modification of deviant cognitions and the enhancement of victim empathy are central components in many treatment programs for sex offenders. There appear to be three broad problems with self-report measures of these factors: variations in the psychometric evaluation of measures; the transparency of items and thus the likely influence of social desirability; and the difficulty of determining which measures are specific to particular types of sex offenders. The aim of this study was to investigate these three issues among child molesters (CMs), and men convicted of sex offences against adults (ASOs). Data were collected from 36 CMs and 31 ASOs and from two comparison groups (33 men convicted of nonsexual offences and 40 nonoffenders from the community), to assess the reliability (internal and test-retest) and validity (discriminant, construct, and face) of measures, the influence of sexual social desirability on responding and the specificity of measures to both sex offender groups. Collectively, the results raise issues related to the assessment of sex offenders that require further investigation. They also have theoretical implications about the relationship between cognitive and emotive processes among sex offenders. PMID- 11501299 TI - Victim-choice polymorphia among serious sex offenders. AB - The victim-choice polymorphia of 178 sexual aggressors divided into six subtypes, incest offenders, pseudoincest offenders, sexual aggressors of familiar children, sexual aggressors of unfamiliar children, sexual aggressors of familiar women, and sexual aggressors of unfamiliar women, was compared. Results showed that sex offenders remained stable in their choice of victim from one offence to another in terms of victim age, victim gender, and aggressor-victim relationship. Subjects characterised by high levels of polymorphia were pseudoincest offenders and sexual aggressors of familiar women, whereas sexual aggressors of both unfamiliar women and unfamiliar children were characterised by low levels of polymorphia. Recommendations regarding how to further refine sex offender typologies are discussed. PMID- 11501300 TI - Comparative data of childhood and adolescence molestation in heterosexual and homosexual persons. AB - In research with 942 nonclinical adult participants, gay men and lesbian women reported a significantly higher rate of childhood molestation than did heterosexual men and women. Forty-six percent of the homosexual men in contrast to 7% of the heterosexual men reported homosexual molestation. Twenty-two percent of lesbian women in contrast to 1% of heterosexual women reported homosexual molestation. This research is apparently the first survey that has reported substantial homosexual molestation of girls. Suggestions for future research were offered. PMID- 11501301 TI - (Serious) sadomasochism: a protected right of privacy? AB - Several adult male members of a sadomasochistic sex club were arrested for violating a U.K. statute passed in 1861--The Offences Against the Person Act. The sex acts spanned a 10-year period and were videotaped. They involved manipulation of the genitalia with hot wax, sand paper, fish hooks, and needles. The sexual interactions were consensual. This analysis tracks the case from trial, to the Court of Appeal, to the House of Lords, and on to the European Court of Human Rights. It examines whether or not these sexual behaviors should be protected under a right to privacy. It contrasts state concerns of bodily harm, albeit consented to, in sex with those incurred in sport. PMID- 11501302 TI - A column study of soil contamination by lead: influence of pH and carbonate content. II. Mathematical model. AB - A mathematical model is used for the interpretation of the results from earlier experimental studies in lab-scale columns on the contamination of a carbonatic soil with lead. Local equilibrium conditions suffice to reproduce the experimental curves for every pH value of the influent contaminant solution and carbonate content of the soils essayed, but heterogeneous contact between the aqueous and solid phase should be included. This heterogeneous contact is responsible for the important tailing effects observed, and is difficult to estimate even for the lab conditions. Then, important uncertainties should be accepted both for risk assessment and in situ remediation feasibility studies. PMID- 11501303 TI - Factors affecting nitrite build-up in nitrifying biofilm reactor. AB - Nitrite build-up in a nitrifying biofilm reactor for direct denitrification from the nitrite stage was investigated. At least three factors were found to influence the nitrite build-up: (i) the relative specific growth rates of Nitrosomonas to Nitrobacter, microNs/microNb in the biofilm; (ii) the relative initial ratio between Nitrosomonas and Nitrobacter on the support surface, (Mao)Ns/(Mao)Nb; (iii) the level of free ammonia, particularly at greater than 0.1 mg N/l that can be inhibitory to Nitrobacter. Results showed that without free ammonia inhibition, the first two factors determined the degree of nitrite build-up. The effect of (Mao)Ns and (Mao)Nb on nitrification kinetics were more significant than the specific growth rates of Nitrosomonas and Nitrobacter. By regulating effects of these three factors, extremely high concentrations of nitrite build-up could occur in a biofilm reactor. This offers a potential for achieving direct denitrification through a nitrate shunt. PMID- 11501304 TI - Influence of different bioparticles on bed expansion characteristics of anaerobic fluidized bed reactors. AB - Bed expansion characteristics and predicting biofilm development and biomass hold up in the anaerobic fluidized bed reactor were investigated. The treatability studies of brewery wastes were carried out in a pilot-scale fluidized bed reactor for a nine months period. The expansion and particle mixing characteristics of a fluidized bed containing different bioparticles is a complicated function of many variables, including hydrodynamics, support characteristics, and biofilm structures. The biological fluidized bed reactors have two types of bed expansion; the first is due to an increase of the superficial velocity and the second is due to the microbial growth in the bed. A new relationship for the porosity of the biological fluidized beds, was developed. Biofilm thickness and bed porosity increased rapidly in the upper part of the bed and the bioparticles become lighter than the clean particles with decreasing effective density. Most bioparticles in the anaerobic fluidized bed reactor (AFBR) were ellipsoidal and the geometric mean diameters were considered as the characteristic diameter. Their surface roughness causes an increase in the drag force acting on the particle. Thus, the expansion coefficient increased from 3.44 to 3.64 as terminal settling velocity decreased from 6.99 to 4.29 cm/s. An almost linear relationship was observed between local porosity and biofilm thickness. The differences between the measured and calculated biomass concentrations varied in the range of 0.4 and 12% (average 4.6%) considering the results from the pilot-scale AFBR. The bed expansion rate of the AFBR, varied from 12.3 to 18.3% whereas, the expansion rate of the clean bed also found between 7.9 and 12.7%. Consequently, the bed expansion rate due to biofilm accumulation on the particles can be well described to vary between 4.4 and 5.6%. PMID- 11501305 TI - Sorption kinetics of lead ions by zeolite tuff. AB - The kinetics of sorption of Pb ions by zeolite tuff treated with NaCl solution was studied using agitated batch laboratory apparatus. The uptake of ions was determined as a function of time at different values of initial Pb ion concentration, zeolite particle size, slurry concentration, solution temperature and pH. The pseudo-second order reaction model was successfully applied to the experimental data with correlation coefficients higher than 0.98. Sorption rates decreased with increase in average particle diameter while it increased with increase in each of the initial Pb ion concentration, slurry concentration, solution temperature and solution pH. Results indicate the importance of chemisorption. PMID- 11501306 TI - The removal of bacteria by modified natural zeolites. AB - The removal effect of natural and modified zeolites containing different heavy metals (Ni2+, Zn2+, Fe3+ and Cu2+) on pure cultures of Escherichia coli and Staphylococcus aureus in a solid medium was evaluated in this work. These experiments were carried out in a continuous mode treating municipal wastewater. Faecal coliform species and Pseudomonas aeruginosa were identified. The rate constants of heavy metal lixiviation were determined using a first order kinetic model. The removal effect of modified natural zeolites in both a solid medium and in continuous mode showed an increased elimination of the bacterial population. The results established a decreasing order of the removal effect as follows: Cu2+ > Fe3+ > Zn2+ > Ni2+. The best performance of columns was obtained for inlet bacterial concentrations below 10(6) cells/100 ml. Most of the identified bacterial species were affected by copper modified zeolites, although Serratia marcescens presented the highest sensitivity and Klebsiella pneumoniae the greatest resistance. PMID- 11501307 TI - Mineralogical and hydrochemical effects on adsorption removal of cesium-137 and strontium-90 by kaolinite. AB - Adsorption characteristics of the nuclides onto kaolinite were investigated by batch experiment under various pH conditions and concentrations of groundwater cations (Ca2+, Mg2+, K+ and Na+) and anions (HCO3-, CO3(2-) and SO4(2-). Adsorption removal of 137Cs and 90Sr by kaolinite greatly increased as the concentration of groundwater cations increased from 10(-5) to 10(-1) M. In contrast, the pH exerted a small effect on the adsorption of 137Cs and 90Sr onto kaolinite. The zeta potential of kaolinite particles showed a negative increase of amphoteric surface charge with increasing pH. The adsorption behavior of 90Sr was also highly dependent on the concentration of bicarbonate. The thermodynamic saturation index indicated that bicarbonate exerts great effect on strontium adsorption by the precipitation of a strontianite (SrCO3) and a change in pH. PMID- 11501308 TI - Feasibility study of a constructed wetland for sewage treatment in a Korean rural community. AB - A pilot study was performed to examine the feasibility of using a constructed wetland system for treatment of sewage in a Korean rural community. The treatment system was a subsurface flow wetland with a loading rate and hydraulic residence time of 6.3 cm/day and 3.5 days, respectively. The wetland system was highly effective in treating the sewage; median removal efficiencies of BOD5 (biological oxygen demand) and TSS (total suspended solids) were about 70.0%, with median effluent concentrations of 24 and 13 mg/L, respectively, for these constituents. However, BOD5 and TSS did often exceed the effluent water quality standards of 20 mg/L. Removal of TN (total nitrogen) and TP (total phosphorus) was relatively less effective and median effluent concentrations were approximately 81 and 8 mg/L, respectively. The treatment system did not experience any clogging or accumulation of organic and inorganic solids during the study, and during winter a substantial amount of removal of BOD5 and TSS continued to occur. Overall, the constructed wetland was judged to be an effective sewage treatment system, but not sufficient by itself to attain regional water quality standards. Therefore, treatment of rural sewage by a wetland system should be followed by another process such as use of the effluent for irrigation of agricultural crops (e.g., rice). PMID- 11501309 TI - Biodesulphurization of coal: mechanism and rate limiting factors. AB - The pyrite sulphur removal from coal by Thiobacillus ferrooxidans was studied in batch reactor. A combination of SEM, IR and XRD was used to study the presence of superficial phases and the changes in solid surface during biodesulphurization. Biodesulphurization was found to be a three-step process. In the first step (0-4 days), direct oxidation of pyrite by bacteria brought about 28% pyritic sulphur removal. Both direct and indirect oxidation contributed to the second step (4-10 days) resulting in 51% pyrite removal. The deposition of elemental sulphur, jarosite and ferric sulphate precipitates in the third step reduced the pyrite availability and ferric iron concentration in the leachate and brought the process of biodesulphurization to an end. PMID- 11501310 TI - Cement-based stabilization/solidification of metal plating industry sludge. AB - This study examines the cement-based Stabilization/Solidification (S/S) technique of sludge produced from a metal plating industry. The sludge samples were characterized in terms of pH and heavy metal content (Cr, Cu, Fe, Ni, Pb and Zn). The leachability of the sludge was estimated using the Toxicity Characteristic Leaching Procedure (TCLP). Two binder mixtures were used for the S/S process, fly ash/cement and zeolite/cement. The weight ratio of the binder mixtures was optimized to achieve the highest strength. The optimum ratio binder mixtures was mixed with sludge samples of different weight ratios and cured for 28 days in order to find the S/S products with the highest strength and the lowest leachability. PMID- 11501311 TI - Degradation of dimethylphthalate by cells of Bacillus sp. immobilized in calcium alginate and polyurethane foam. AB - A Bacillus sp. which is capable of degrading dimethylphthalate (DMP) was immobilized in calcium alginate and polyurethane foam for efficient and long term degradation of DMP. Freely suspended cells (10(12) cfu ml-1) degraded a maximum of 20 mM DMP. Whereas, alginate-(10(12)cfu g-1 beads) and polyurethane foam entrapped (0.34 x 10(6-9) cfu g-1 foam cubes) cells degraded a maximum of 40 mM DMP within 12-15 days of incubation. Polyurethane foam-entrapped cells degraded 30 mM of DMP at 4 days and alginate-entrapped cells degraded within 10 to 12 days of incubation irrespective of the cell population. When the initial concentration of DMP increased to 50 mM, the DMP degrading ability of the immobilized cells was not increased even after 20 days. Repeated batch cultures by alginate-entrapped cells with initial 35 mM DMP loading could be reused for a maximum of 20 cycles. However, the degradation rate was gradually decreased when the beads were reused for more than 15 cycles. On the other hand, the foam-entrapped cells, with the same initial DMP loading there was no decrease in DMP degrading ability and could be reused for more than 20 cycles. The packed bed reactor with alginate-entrapped cells (1 x 10(10-12) cfu g-1 bead) could be continuously operated for 7-8 days with an initial 25 mM DMP at a flow rate of 50 ml h-1. Whereas, the polyurethane foam-entrapped cells (1 x 10(6-9) cfu g-1 foam cubes) could be operated continuously for more than 90 days with the same initial DMP loading at a flow rate of 100 ml h-1. Thus the enhanced degradation of DMP could be achieved by immobilizing the cells of Bacillus sp. in calcium alginate and polyurethane foam as compared to that of freely suspended cells. PMID- 11501312 TI - Biodegradation potential of photocatalyzed surfactant washwater. AB - Enhanced release of hydrophobic compounds from a soil matrix can be achieved by use of soil-washing or soil-flushing using various surfactants. However, the surfactants used in achieving the desorption of organic contaminants may also cause a problem in subsequent removal/disposal of these contaminants. UV radiation in the presence of TiO2 as a pre-treatment step to achieve initial (or partial) breakdown of naphthalene and Sodium Dodecyl Sulfate (SDS) using batch experiments indicated that 56% to 88% naphthalene degradation occurred within 30 minutes to one hour. Preliminary results on the estimate of the batch aerobic biodegradation potential of photocatalyzed washwater containing naphthalene and SDS suggested that SDS was the major carbon and energy source for an activated sludge enrichment culture and an enrichment culture obtained from microorganisms at a contaminated site. Continuous-flow stirred tank reactors (CSTRs) with with a solids retention time (SRT) of 4 days were not effective, but an SRT of 8 days was successful in biodegrading the naphthalene and surfactant. These results indicated that photocatalytic treatment as a pre-treatment step followed by a biodegradation step may offer potential in cleaning up surfactant washwaters containing organic contaminants. PMID- 11501313 TI - Automated sampling of stormwater runoff in an urban watershed, north-central Texas. AB - Automated samplers were used to collect urban runoff in Pecan Creek, Denton, Texas. Runoff from four storm events was sampled at four stations. Concentrations of some heavy metals (cadmium and arsenic), as well as pesticides (atrazine and diazinon), exceeded maximum contaminant levels for drinking water, but were within ranges typical for urban runoff. Calcium and phosphorous concentrations were high compared to previous studies, which was attributed to soils, building materials, and fertilizer applications in the study area. First flush samples were more concentrated than composite samples for most of the constituents analyzed, but statistically different for only five constituents. Concentrations of four constituents differed significantly among stations, and six differed significantly among storms, due to variations in land use, channel composition, and storm conditions. Overall, the automated samplers were an effective way to sample urban runoff in Pecan Creek. PMID- 11501314 TI - A study of heavy metal pollution in Lake Victoria sediments by Energy Dispersive X-Ray Fluorescence. AB - Sources of heavy metal pollution of Lake Victoria is of interest due to its economic and domestic implication in East Africa. Sediments from the shore of Lake Victoria and from some streams flowing into the lake have been analyzed for their heavy metal content using Energy Dispersive X-Ray Fluorescence (EDXRF) analysis. The samples were collected from seventeen different locations around the Municipality of Mwanza divided into three zones according to their activities. The results show that sediments from Mwanza North, which is least in anthropogenic activities, contains Cr and Co in addition to metals of terrestrial origin (K, Ca, Ti, Mn, Fe, Rb, Sr, Y, and Zr), samples from Mwanza Central, which is the town center, contains high concentrations of V, Cu, Zn and Pb, and samples collected from Mwanza South, which is the industrial area, contains the highest concentrations of V, Cu, Zn, As and Pb. It is concluded that the industrial and sewage wastes discharged into the lake are the main sources of the heavy metal contamination of Lake Victoria. Further studies will be required to assess detriments of these pollutants to human and aquatic life before policy for the factories responsible is put in place. PMID- 11501315 TI - Electrochemical remediation of trichloroethene-contaminated groundwater using palladized iron oxides. AB - The objective of this study is to develop electrochemically-enhanced dechlorination of trichloroethene (TCE) using palladized iron oxides minerals for ex situ remediation of contaminated groundwaters. A bench-scale column packed with the palladized iron oxide media connected to a cathode and an anode embedded in a carbon pad was prepared for flow through column tests. Contaminated groundwaters with about 14-16 mg/L TCE were passed from the cathode side to the anode side of the column while the system was supplied with direct current. All of the TCE in the groundwater was dechlorinated even after 300 pore volumes were passed. Furthermore, intermediate reaction products, dichloroethene isomers and vinyl chloride, were not detected in the treated water. PMID- 11501316 TI - A simple electrolyte for determination of small cations in natural waters by capillary electrophoresis. AB - Three simple electrolyte systems were tested for determination of four alkali and alkaline earth cations (K+, Na+, Ca2+, Mg2+) in aqueous sulfate solutions. The separation was achieved in a system of 5 mM 4-aminopyridine and 5% v/v methanol with indirect UV detection at 214 nm, in which the electrolyte pH was adjusted to be 4.30 by adding 1 M glycolic acid. Four cations were well separated within 5 min at an applied voltage of 15 kV. Linear relationships of the calibration curves were obtained up to 50 ppm for all four cations. To evaluate this electrolyte system, the determination of these cations was also conducted for real rain waters. PMID- 11501317 TI - Ultrasonic destruction of chloroform and carbon tetrachloride in aqueous solution. AB - The sonochemical destruction of binary mixtures of chloroform and carbon tetrachloride in aqueous solution at 30 kHz has been studied. The influences of initial concentration, bicarbonate/chloride, and acoustical intensity on the destruction rates have been observed. It was found that the destruction of the two compounds appeared to follow pseudo-first-order reaction kinetics with the current experimental concentration range. The rate constant of chloroform for initial concentration as high as 100.4 mg/L decreased compared to initial concentration 36.13 micrograms/L, when the rate constants of carbon tetrachloride hardly changed. However, their destruction rates increased with increasing acoustical intensity and were independent of addition of bicarbonate or chloride. The resulting yield of ionic product (Cl-) for chloroform was determined. PMID- 11501318 TI - Environmental assessment of a site contaminated by organic compounds. AB - This paper presents a study on environmental assessment of an abandoned industrial area located in central Italy. Main production was refractory materials and compounds for treatment of industrial wastewater. The present work deals with a methodology for development of a sound sampling design, chemical characterization of soil samples, definition of the degree of site contamination according to law limits and evaluation of the fate and transport of contaminants by EPA simulation model (VLEACH 2.2a). Results indicate that toxic compounds (polycyclic aromatic hydrocarbons and plasticizers) are uniformly distributed in the contaminated site and only in one sampling point their concentrations exceed law limits. Modeling results confirm that contaminants migration to groundwater can be excluded, addressing for a site remediation limited to the surface layer. PMID- 11501319 TI - The effectiveness of municipal sewage sludge application on the stabilization of Pb, Zn, and Cd in a soil contaminated from mining activities. AB - The effectiveness of municipal sewage sludge for the stabilisation of Pb, Zn and Cd in a heavily contaminated soil was evaluated by performing pot experiments on soil-sludge mixtures. The soil sample originated from the Montevecchio mining district, Sardinia, Italy, and presented high Pb, Zn and Cd content, as well as US EPA TCLP solubility values for Pb and Cd, which exceeded the respective regulatory limits. Sewage sludge application increased the soil pH. Stabilisation experiments showed that 10% w/w sewage sludge addition effectively reduced Pb and Cd solubilities below the TCLP regulatory limits. At the same addition rate, the EDTA extractable fraction of Pb, Zn, Cd in the treated soil was reduced by 12, 47 and 50% respectively compared with the untreated sample. The five-stage sequential extraction procedure applied on the untreated and treated soil samples, showed a remarkable shift of the metals towards more stable forms. The reducible fractions of Zn and Cd and the residual fraction of Pb were increased by 12, 20 and 18% respectively, while a corresponding decrease in the mobile fractions (exchangeable and carbonate) occurred which accounted for 14, 23 and 25% respectively. PMID- 11501320 TI - Biosorption of heavy metals from landfill leachate onto activated sludge. AB - The removal of various heavy metals was studied when activated sludge was exposed to heavy metals in landfill leachate. Batch uptake tests were conducted for this purpose. Adsorption was the main mechanism of removal when biomass was contacted with heavy metals. Activated sludge had a high biosorption capacity and equilibrium was reached in a short time with respect to copper, iron, manganese, zinc and chromium. Adsorption isotherms were generated for those heavy metals and the Freundlich constants were calculated. Among the metals studied, manganese became very concentrated on activated sludge with time. PMID- 11501321 TI - Air sparging tracer test to investigate the flow path. AB - The paper presents a useful method for investigating the flow path of the injected air, estimating the radius of influence, and assisting the design of well spacing of sparging wells. The method applies both tracer and modeling approach. A field test using helium as a tracer was conducted on an air sparging pilot. A 3-dimensional, 3-phase compositional model was utilized to simulate the air sparging process at the pilot. The model was first calibrated with the tracer concentrations observed in the field test at various locations between the air sparging well and the soil vapor extraction well. The calibrated model was then used to predict the flow path of the injected air and to simulate the water saturation distributions in the aquifer. The steady-state water saturation distributions was found useful for estimating the radius of influence of air sparging and for designing the well spacing of sparging well. PMID- 11501322 TI - TNF alpha blockade in rheumatoid arthritis--time for caution or enthusiasm? PMID- 11501323 TI - Patient empowerment stroke--a strategy for Scotland. PMID- 11501324 TI - Myocardial infarction in men and women under 65 years of age: no evidence of gender bias. AB - We examined short and long term outcomes of MI in a consecutive series of 169 men and 50 women who were followed for an average of 3.5 years. Similar percentages of men and women were admitted to medical intensive care, received in-patient cardiac rehabilitation, quit smoking at one year, were still smoking, were taking a lipid lowering drug or had returned to work at one year, underwent coronary angiography at 3.5 years or had died by 3.5 years. The lack of gender difference in outcome may reflect an absence of gender bias in the management of men and women with MI in southwest Scotland. PMID- 11501325 TI - Reputation and the legibility of doctors' handwriting in situ. AB - The consequences of failed communication can be as disastrous and doctors have a reputation for illegibility. Previous research supports this opinion but little work has been done to assess doctors' legibility in situ. Our study evaluates if doctors deserve their reputation and investigates how legibility is affected by the time taken to write. Sets of in-patient hospital notes were selected at random. The first written entry by a doctor and a nurse in the current admission were analysed. In addition to this, 10 doctors and 10 nurses, unaware of the true nature of the study, wrote out lists of words and the time taken to do the task was recorded. The doctors' handwriting was significantly less legible (p = 0.010) and they wrote significantly quicker (p = 0.005). However a small minority of the doctors was responsible for the majority of illegible words written by that group. PMID- 11501326 TI - Playing the waiting game ... the asymptomatic patient with recurrent ovarian cancer detected only by rising Ca125 levels. AB - The recurrence of ovarian cancer is usually heralded by an increase in the blood level of the tumour marker Ca125 but most oncologists are reluctant to initiate second-line chemotherapy before there is clinical or radiological evidence in addition to the increasing tumour marker. Thus patients have to spend an indefinite period of time, which can be as long as a year or more, knowing that their cancer has returned, but that treatment will be delayed until they have symptoms. For patients, this situation requires a change of outlook and an abandonment of the idea that early treatment is the best option. A dilemma exists for both oncologists and patients over the best time to begin aggressive treatment in this situation. This article explores some of the arguments for and against early treatment, in the hope that it might provide some insight into patients' concerns, and help patients to enjoy their symptom-free and treatment free period of recurrence with some measure of tranquillity. PMID- 11501327 TI - Mortality during the winter flu epidemic--two cases of death associated with self medication. AB - We report two cases of mortality associated with the recent winter influenza outbreak. Both cases were associated with self-medication. In one case an elderly lady died from haemorrhagic duodenitis induced by over the counter ibuprofen. In the second case the lady died from the consequences of exceeding the recommended doses of paracetamol by combining doses of the generic product with proprietary flu-remedies and Tylex (paracetamol and codeine). PMID- 11501328 TI - Free wall rupture following rescue angioplasty and intravenous abciximab for failed thrombolysis. AB - Early thrombolytic therapy reduces the risk of cardiac rupture but delayed thrombolysis may increase this risk, despite improving overall survival. The mechanism appears to be related to both unsuccessful early reperfusion and haemorrhagic transformation following delayed reperfusion. The effect of antiplatelet therapy with glycoprotein IIb-IIIa receptor blockers (abciximab) on cardiac rupture is unknown. It is possible that they may contribute to cardiac rupture by promoting haemorrhagic transformation of the infarcted area. In this report we describe a 57 year old man who underwent emergency coronary angioplasty and stenting following failed thrombolytic therapy for an acute anterior myocardial infarction. A suboptimal result was obtained which necessitated an intravenous bolus of abciximab followed by an infusion. He abruptly developed electromechanical dissociation. Echocardiogram confirmed pericardial tamponade and a pericardial drain was inserted but the patient could not be resuscitated. Postmortem examination confirmed a large transmural rupture of the infarcted anterior wall which had undergone haemorrhagic transformation. PMID- 11501329 TI - Two sons of Paisley and the birth of academic pathology in Scotland. PMID- 11501330 TI - Proactive safety surveillance. AB - Growth in health information systems presents opportunities to enhance postmarketing safety surveillance of medical products. Spontaneous suspected side effect reports provide the foundation, but we need to 'proactively' improve their quality and our strategies to seek signals. In our more familiar 'reactive' mode, we examine hypotheses from inquiries or publicity. Such responsive evaluations remain essential but may miss latent information on unsuspected risks. Efficient techniques to disclose hidden clusters and associations may emerge through adaptation of approaches from industrial quality control and other disciplines. Data-driven techniques like exploratory analysis, control charts, and time series modeling may help in sifting through accumulated data and in screening consecutive submissions to discern hints of new product hazards or of more specific understanding about previously identified potential side effects. We also need to cultivate non-spontaneous data for hypothesis generation as well as testing, the systematic epidemiologic evaluation of questions and concerns. This hypothesis testing function will assume greater importance if proactive safety surveillance methods yield larger numbers of putatively positive findings. Whether from spontaneous reports or other sources, signals that could have arisen by chance alone usually represent only clues to potential hazards until or unless they can be verified through independent studies. PMID- 11501331 TI - Signal generation and clarification: use of case-control data. AB - Case-control surveillance systems are useful for 'signal' generation, i.e., signaling potential previously unidentified adverse effects of drugs. Two systems currently in operation, the Slone Epidemiology Unit's Case-Control Surveillance and the Birth Defects Study, have monitored drug effects since 1976. With extensive information on the diagnoses and covariates, the systems have the capacity to carry out in-depth analyses in which the outcome measure is more specifically defined and in which confounding is controlled, thus reducing the possibility of false alarms. PMID- 11501332 TI - Risk management of marketed drugs: FDA and the interface with the practice of medicine. PMID- 11501333 TI - Drug safety: can simple interventions be effective in a complex world? PMID- 11501334 TI - Contraindicated medications dispensed with cisapride: temporal trends in relation to the sending of 'Dear Doctor' letters. AB - PURPOSE: 'Dear Doctor' letters alert the prescribing community of drug labeling changes that contain new contraindications, warnings, adverse reactions, and precautions. There has been little assessment of the impact of these letters. We quantified the impact of two 'Dear Doctor' letters concerning interactions between cisapride and a series of drugs. A letter in 1995 described a risk of prolonged QT intervals and serious ventricular arrhythmia in patients who received macrolide antibiotics and imidazole antifungals in conjunction with cisapride. A June 1998 letter that expanded the list of contraindicated comedications had wider distribution than an earlier one, was accompanied by substantial Internet and media coverage, and was complemented by an effort to inform large pharmacy dispensing information organizations of the warnings against concurrent use of the named drugs. METHODS: Health plan members with one or more outpatient pharmacy claims for cisapride during the period 1 January 1995 through 31 May 1999 were identified among members of a large New England health insurer. A retrospective review of concurrent and nearly concurrent dispensings of cisapride and contraindicated comedications was undertaken in the automated pharmacy claims data using both graphical and statistical time-series analysis. We tabulated by month the fraction of cisapride dispensings that occurred in close temporal relation to dispensings of contraindicated comedications. Codispensings that occurred on the same day were taken as the most direct measure of prescriber responsiveness to the letters. Codispensings that occurred in windows of plus or minus 2 weeks (29 day window) and plus or minus 4 weeks (57 day window) were taken as measures of possible simultaneous consumption. Among overlapping dispensings, we counted the proportion dispensed by the same pharmacy. Time series regression analysis of secular, seasonal, and step-effects was conducted. RESULTS: There was a steady decline in codispensing of cisapride and contraindicated medicines, and a pronounced seasonal effect, arising principally from the seasonal use of macrolide antibiotics. Against this background, the isolated Dear Doctor letter of October 1995 had no discernible effect on prescribing practices. The 1998 letter and surrounding activity, by contrast, were followed by a 66% decline in same-day dispensings and a smaller, but still pronounced decline in dispensings in the wider time windows. For most codispensings of contraindicated medications with cisapride, both medications came from the same pharmacy. CONCLUSIONS: Publicity and direct intervention with dispensing pharmacies may be an important supplement to Dear Doctor letters when the goal is to eliminate the codispensing of drugs that should not be taken together. PMID- 11501335 TI - Antihypertensive drug prescription trends at the primary health care centres in Bahrain. AB - PURPOSE: To determine the antihypertensive drug prescribing pattern by primary care physicians in patients with uncomplicated essential hypertension; to identify whether such pattern of prescription is appropriate and in accordance with international guidelines for pharmacotherapy of hypertension; and to estimate the impact of such prescriptions on cost of treatment. METHODS: A prescription-based survey among patients with uncomplicated essential hypertension was conducted in seven out of a total of 18 health centres in Bahrain. The relevant data for our study was collected using cards, designed for chronically-ill patients. RESULTS: A total of 1019 male and 1395 female (62.9%) out of 3838 of the study population were on monotherapy, whereas 596 male and 828 female (37.1%) were on antihypertensive combination therapy. Among the monotherapy category, the various antihypertensive drugs used were as follows: beta-blockers (58.8%), angiotensin converting enzyme (ACE) inhibitors (14.2%), calcium channel blockers (11.1%), diuretics (8.1%) and alpha-methyldopa (7.0%). With respect to overall utilization pattern, beta-blockers were the most frequently prescribed (65.5%), diuretics ranked second (27.4%), followed by ACE inhibitors (20.6%), calcium channel blockers (19.9%) and alpha-methyldopa (8.5%). Within each class of antihypertensives used, the most frequently used individual agents were as follows: (a) among beta-blockers 97.7% used atenolol; (b) among the diuretics, indapamide (35.4%), hydrochlorothiazide (HCTZ) (32.7%), HCTZ in combination with triamterene (25.7%), and chlorthalidone (4.6%); (c) among the ACE inhibitors, captopril (44.9%), enalapril (29.7%), and lisinopril (19.0%); (d) among the calcium channel blockers, nifedipine (98.2%). Significant age- and gender-related differences in prescribing patterns were seen. Short-acting nifedipine monotherapy was inappropriately prescribed in a significant number of patients above the age of 50 years. ACE inhibitors accounted for approximately two-thirds of the total antihypertensive drug expenditure, although these drugs represent only one-fifth of overall antihypertensives used. There is a trend towards excessive use of expensive thiazide-like diuretics such as indapamide which seems to be unjustifiable practice, particularly in a study population free from diabetic hypertensive patients. CONCLUSIONS: The general pattern of antihypertensive utilization appears to be in accordance with the guidelines of WHO and the Joint National Committee issued in the 1990s. The trends of prescribing of antihypertensives were in favour of conventional ones such as the beta blockers and diuretics, and the introduction of newer classes of antihypertensives had a generally minimal impact on the prescribing profile. Almost two-thirds of the patients were treated with monotherapy. A disproportionately large percentage of antihypertensive drug cost was due to overt use of ACE inhibitors, and indapamide, instead of thiazide diuretics. The use of short-acting calcium channel blockers especially in the elderly is unjustifiable. PMID- 11501336 TI - Collaborative study on drug use in pregnancy: the results of the follow-up 10 years after (Novi Sad Centre). AB - An international multicenter study entitled Collaborative Study on Drug Use in Pregnancy (DUP) was initiated in 1987 by the Mario Negri Institute (Milano) and co-sponsored by the WHO Regional Office for Europe (Copenhagen). The study covered the use of medicines during pregnancy, labour and postpartum period. The Novi Sad centre contributed with 296 questionnaires. The aim of the study was to evaluate the pharmacotherapy by means of comparison, to identify problems in a particular settings and to plan actions toward improvement. The prescribing of some medicines in the Novi Sad center was found to be at the extreme, being among the lowest in the use of iron preparations during pregnancy and among the highest in the use of ergot alkaloids in the postpartum period. Educational efforts have been made to change this practice after the conclusion of the study in 1989. Ten years later, in 1999, the same questionnaire was applied to 100 women in the postpartum period in the same nursery, the Department of Obstetrics and Gynecology in Novi Sad. The results have shown a distinct improvement with the increase of iron prescribing during pregnancy and almost complete abandonment of postpartum use of ergot alkaloids. In the second study period some other inconsistencies with accepted therapeutical principles were identified. For example, although the use of analgesics fell from 22.3 to 8%, the selection changed towards the high use of metamizole (noraminophenazone) and no use of paracetamol (acetaminophen). It was concluded that international pharmacoepidemiological studies such as DUP, could substantially contribute to the improvement of pharmacotherapy if followed by appropriate actions in changing the prescribing behaviour of the physicians and information directed to the target population. PMID- 11501337 TI - An audit of the use of antirheumatic drugs in a north Indian referral hospital. AB - OBJECTIVE: This study was conducted with the aim of auditing the pattern of use of antirheumatic drugs in a tertiary care hospital in Northern India. METHODS: The study was carried out in 1000 patients recruited sequentially from the clinic for a period of 1 year (January to December 1999). Patient data such as age, sex, income, family size, diagnosis, duration of illness, drugs prescribed/duration, adverse drug reaction were noted and used to calculate core drug use indicators and pattern of drug use. RESULTS: The pattern of drug use was in accordance with the standard practices followed internationally. In rheumatoid arthritis the most common prescriptions were for non-steroidal antiinflammatory drugs (NSAIDs) alone, followed by the combination of NSAIDs, disease modifying agents (DMARDs) and steroids. Of the NSAIDs diclofenac was the most frequently prescribed drug, while chloroquine was the most commonly used DMARD. The most commonly seen adverse drug reactions were gastritis, Cushings syndrome and decreased visual acuity. CONCLUSION: In conclusion, this study has demonstrated that in this clinic, the pattern of use of antirheumatic drugs follows standard guidelines. PMID- 11501338 TI - Is hormone replacement therapy protective for venous ulcer of the lower limbs? AB - PURPOSE: Estrogen has been found to increase the risk of deep vein thrombosis (DVT), which is associated with venous ulcers. A matched case-control study was therefore performed to quantify the effect of hormone replacement therapy (HRT) on the occurrence of venous ulcers of the lower limbs. METHODS: Women presenting to a participating vascular surgery department between January and December 1997 with a first open venous ulcer served as cases. Controls were sampled among women with sub-acute conditions such as skin problems, cold, headache/migraine, sore throat and mild ear infections, and were matched on referral site and age (+/- 5 years). Subjects were eligible for this study if they were postmenopausal, and excluded if they had non-palpable pedal pulse or any chronic active diseases. The study consisted of an interviewer-administered risk factor questionnaire. Conditional multivariate logistic regression analyses were performed adjusting for history of DVT and unbalanced distributions in important covariates. RESULTS: Forty-four cases and 80 matched controls were recruited. The mean age of participants was 64 years. HRT users were significantly thinner (27% with body mass index (BMI = (kg/height (m2)) > or = 27 vs. 41%, p = 0.04) and less likely to have a history of DVT (12 vs. 33%, p = 0.02) and varicose veins (VV) (52 vs. 69%, p = 0.04) than non-users. A matched univariate logistic regression analysis was performed to quantify the effect of HRT on venous ulcers and generated an odds ratio (OR) of 0.14 with 95% confidence interval of (0.04, 0.49)95%: adjusting for prior DVT resulted in an OR of 0.29 (0.09, 0.93)95%. When adjusting for BMI, history of VV, exercise, education, and smoking, the OR observed was 0.03 (0.01, 0.13)95%: further adjustment for prior DVT gave an OR estimate of 0.06 (0.01, 0.50)95%CI. CONCLUSIONS: Our study demonstrates that HRT has a protective effect on the occurrence of venous ulcers of the lower limbs. Adjusting for prior DVT moves the estimate closer to the null. DVT is a strong contraindication for HRT and therefore must be considered in order to eliminate confounding by contraindication. PMID- 11501339 TI - Report on the 12th Annual Scientific Meeting of the UK Drug Utilisation Research Group, Friday 15 December 2000, the Royal Society of Medicine, London. PMID- 11501340 TI - The case-crossover and case-time-control designs in pharmacoepidemiology. PMID- 11501341 TI - The loss of assets to nursing homes can be avoided by proper planning. PMID- 11501342 TI - Interaction between apolipoprotein-E and angiotensin-converting enzyme genotype in Alzheimer's disease. AB - Both apolipoprotein-E (apo-E) epsilon 4 allele and angiotensin-converting enzyme (ACE) deletion (D) polymorphism have been associated with a high risk for coronary heart disease. Increased frequency of the epsilon 4 allele has also been reported in patients with late-onset of familial and sporadic Alzheimer's disease (AD). The primary aim of this study is to examine the possible relationship between the ACE gene polymorphism and AD. The second aim of this study is to explore the relation of the ACE and apo-E genotypes with AD. Polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and agarose gel electrophoresis techniques were used to determine the apo-E and ACE genotypes. The frequencies of ACE D and ACE insertion (I) allele among AD patients and controls were 55.7 percent versus 44.2 percent and 51.7 versus 48.2 percent, respectively. Apo-E allele frequencies in the AD group for epsilon 2, epsilon 3 and epsilon 4 were, 1.7 percent, 96.5 percent, and 1.7 percent, respectively. The apo-E allele frequencies of healthy groups for epsilon 2, epsilon 3 and epsilon 4 were 1 percent, 56 percent, and 1.7 percent, respectively. In conclusion ACE D and apo epsilon 4 allele were found to be more frequent in patients with Alzheimer's disease than in the control group. PMID- 11501343 TI - Insight in Alzheimer's patients: results of a longitudinal study using three assessment methods. AB - Three direct measures of dementia insight were administered to 20 participants in a longitudinal Alzheimer's disease (AD) rehabilitation research project and to subsets of these participants that completed one (N = 19), two (N = 12), and three (N = 6) years of program participation. The measures were: (1) responses to a discourse prompt question about AD (ADPQ); (2) endorsements of seven items on the Geriatric Depression Scale (GDS) about the effects of dementia (separate analyses were done for two of the seven items that related specifically to memory and thinking); and (3) a sentence-completion exercise. Responses to measures 1 and 2 and the subset of 2 were quantified, tracked over time, and subjected to correlational analyses with age, Mini-Mental State Exam (MMSE) score, and depression, as measured by total GDS score, and with each other. MAJOR FINDINGS: There were no decreases in insight from baseline to year 1, 2, or 3, as measured by free responses to the AD prompt question. There was a significant decline in insight from baseline to year one on the GDS measure, but no change from year 1 to year 2 and a return to baseline level at year 3. There was no correlation between insight and baseline age, between insight and MMSE score at any time point, between MMSE score and depression, as measured by total GDS score, or between MMSE score and depression score, except for the year 3 completers, where depression score was negatively correlated with MMSE score at year 3 only. GDS insight and ADPQ scores were not correlated. Several participants that showed no insight on the quantified measures did so on the sentence completions. PMID- 11501344 TI - An overview of AIDS dementia complex. AB - The major neurological complications associated with HIV infection include cognitive, behavioral, and motor disturbances, which may range in severity from subtle, mild cognitive deficits to the clinical syndrome referred to as HIV associated dementia or AIDS dementia complex (ADC). As with Alzheimer's type dementia, caregivers for people with HIV/AIDS have the overwhelming and burdensome task of caring for someone with deteriorating cognitive abilities, increasing physical debilitation, and changes in personality. This article describes ADC as well as some of the similarities and differences from Alzheimer's type dementia, and offers some special considerations for older adults and HIV. PMID- 11501345 TI - The employee victim of violence: recognizing the impact of untreated psychological trauma. AB - The impact of psychological trauma in the workplace often goes unaddressed. The untreated aftermath of these critical incidents may manifest itself in various states of anxiety, depression, substance use disorders, and even subsequent violence by the victims. This paper reviews common presentations of untreated traumatic events and provides suggestions for outreach to untreated employee victims as well as basic prevention strategies to reduce the risk of additional episodes of violence and enhance safety in health care facilities, including long term care (LTC) and special-care units (SCU) for dementia patients. PMID- 11501347 TI - The agitation-activity interface of people with dementia in long-term care. AB - The purpose of this study was to examine agitated behaviors of people with dementia relative to time and activity variables. This descriptive study involved a sample of 420 units of 30-minute time periods collected in a long-term care setting. The highest agitation scores occurred during the evening (F = 3.93, p = 0.009). Agitation scores were significantly higher when the same level of activity was sustained for 1.5 hours or longer (p = 0.000). This study suggests that future examination is needed of an intervention to control activity schedules so that there is a balance between sensory-stimulating and sensory calming activities. PMID- 11501346 TI - A day care program and evaluation of animal-assisted therapy (AAT) for the elderly with senile dementia. AB - We conducted a survey to clarify the evaluation methods of animal-assisted therapy (AAT) for the elderly with senile dementia in an adult day care center. AAT was implemented for a total of six biweekly sessions. The AAT group consisted of seven subjects and the control group numbered 20 subjects. In a comparison between Mini-Mental State Exam (MMSE) scores at baseline and those measured three months later, the average MMSE score before AAT (baseline) was 11.43 (+/- 9.00), and three months later it was 12.29(+/- 9.69). In the AAT group, the average score on Nishimura's Activities of Daily Living (N-ADL) at baseline was 28.43(+/- 14.00), and after ATT it was 29.57(+/- 14.47). In the AAT group, the average baseline score on behavioral pathology of Alzheimer's disease (Behave-AD) was 11.14(+/- 4.85), and three months after AAT it was 7.29(+/- 7.11) (p < 0.05). In the control group, the average baseline score was 5.45(+/- 3.27) and three months later it was 5.63(+/- 3.59). The evaluation of salivary CgA, as a mental stress index, showed a decreasing tendency in the AAT group. Our findings demonstrate the usefulness of using several methods for evaluation of the changes in patients given AAT. PMID- 11501348 TI - Abandonment fears of persons with Alzheimer's disease attending adult day care. AB - By using participant observation and staff interviews, we examined the phenomenon of abandonment fears in persons with Alzheimer's disease who were frequent attendees of an adult day care center. Two women, who most frequently expressed fears of abandonment, were the primary participants of the study. Their fears stemmed from two sources: contextual confusion, centered on their inability to grasp the concept of attending an adult day care center, and imagined barriers between them and their caregivers. One of the women tended to "recruit" the other woman into her world of insecurity, and hence their friendship actually may have had a detrimental effect on each other's days at the center. Staff members discussed the issue with reluctance and indicated that methods to attempt to change the behavior would be futile. PMID- 11501349 TI - Using external memory aids to increase room finding by older adults with dementia. AB - When nursing home residents with dementia are unable to locate their own rooms, it often creates problems for staff and other residents. The impact of placing two external memory aids outside participants' bedrooms was evaluated by using a multiple-baseline design experiment. Three female special care unit (SCU) residents with severe Alzheimer's disease (MMSE = 5.7) participated in the study. Results showed that a combination of a portrait-type photograph of the participant as a young adult and a sign stating the resident's name increased room finding by over 50 percent with all three participants. Room finding stabilized at 100 percent accuracy for all participants within a few days of implementing the environmental intervention. PMID- 11501350 TI - Statement on influenza vaccination for the 2001-2002 season. PMID- 11501351 TI - Report on the Cabin Health Conference 2001, 17-18 May, Geneva, Switzerland. PMID- 11501352 TI - Vectors of diseases hazards and risks for travellers: Part I. PMID- 11501353 TI - The role of early radiological studies after gastric bariatric surgery. AB - BACKGROUND: The authors investigated early radiological findings after gastric surgery for morbid obesity to evaluate their usefulness in avoiding complications or facilitating treatment. MATERIAL AND METHODS: 413 patients underwent gastric bariatric surgery: 327 had vertical banded gastroplasty (VBG), 55 Roux-en-Y gastric bypass (RYGBP), 22 adjustable silicone gastric banding (ASGB), and 9 biliopancreatic diversion (BPD). A radiological upper gastrointestinal investigation employing water-soluble contrast medium was performed in each patient between the 2nd and 8th postoperative day. Several techniques were employed to assess different radiological findings related to the anatomic modifications after the bariatric surgery. RESULTS: In VBGs, delayed emptying was found in 10 patients (3%), gastric leak in 3 patients (0.9%), vertical suture breakdown in 1 patients (0.3%), and a wide pouch in 4 patients (1.2%). In RYGBP, a leak was detected in 2 patients (3.6%), delayed emptying in 2 (3.6%), and a wide pouch in 5 (9.1%). ASGB required band enlargement for stomal stenosis in 6 patients (27.2%). Temporary delayed emptying from stomal stenosis was also observed in 2 BPDs (22.2%). Overall complications were 35/413 (8.2%). Two cases of gastric leak after VBG were reoperated. Stomal stenosis after ASGB were treated by percutaneous band deflation; other cases were medically treated until complete healing. CONCLUSIONS: Early radiological study after gastric bariatric surgery is advisable, since it detected postoperative complications (gastric perforation, stomal stenosis, etc.) and modified the clinical approach. As the interpretation of these radiographs is often difficult, involving different projections or patient's positions or other technical managements, surgeons and radiologists must interact and be knowledgable. PMID- 11501354 TI - Partnership, sexuality, and sexual disorders in morbidly obese women: consequences of weight loss after gastric banding. AB - BACKGROUND: A study was performed to determine what consequences surgery for morbid obesity has on sexual attitudes and partnership in obese female patients. METHOD: Semi-structured interviews concerning sociodemographic data, sexuality and relationship were conducted on 82 female patients preoperatively and at least 1 year postoperatively. RESULTS: Physical appearance played the main role in the decision to undergo weight reduction surgery in only 17% of the study patients. Postoperatively, half of the patients were satisfied with their physical appearance 1 year after surgery, the other half not. Preoperatively, 44% of the patients stated that sexuality with their partners was satisfying and the frequency of sexual intercourse was regular. Postoperatively, 63% of the patients stated that they enjoyed sex more, compared with 12% of the patients who enjoyed sex less than before surgery. Postoperatively, 20% of the patients reported that partnership had changed positively, 10% negatively. CONCLUSIONS: The results indicate that many of the sexual problems in obese individuals are the result of an underlying lack of self-esteem, unsatisfactory relationships, or collective stigmatization of obese individuals. Binge eating, often found in morbidly obese patients, seems to be less the result of or compensation for sexual problems but is more likely to stem from other psychosocial or psychological problems. PMID- 11501355 TI - Adaptability and compliance of the obese patient to restrictive gastric surgery in the short term. AB - BACKGROUND: Low compliance, as well as recurrent vomiting, are often pointed out as the main causes of both postoperative technical complications and poor weight loss. METHODS: 50 patients were examined. All underwent preoperative psychological assessment through an intensive clinical interview. Between October 1999 and January 2000, all the patients underwent vertical banded gastroplasty (VBG). Excess Weight Loss (EWL) was followed, and the number of vomiting episodes was recorded at 1, 3 and 6 months after surgery. RESULTS: 3 different groups of patients could be distinguished: 1) those with no vomiting and good weight loss; 2) those with sporadic vomiting and fairly good weight loss; and 3) those with frequent vomiting and poorer weight loss. However, this last group of patients was not homogeneous and should be divided into 2 subgroups: 1) patients who lose weight, while vomiting; 2) patients who do not lose weight, while vomiting. CONCLUSIONS: The experience of vomiting is a crucial cross-roads. The influence of psychosocial factors was noticed in all patients. The absence of psychosocial stressors, together with realistic expectations and a strong motivation to change, should be considered as reliable and replicable predictors of success. However, even those patients with accentuated psychological difficulties were able to succeed in coping with them, and achieved a good weight outcome, with good effects on the psychic side. Therefore, the evidence of psychological disorders cannot be taken as an absolute criterion of selection/exclusion of candidates for the operation. Accurate presurgical education and postoperative psychological support are likely to increase the patient's compliance and the percentage of successful cases. PMID- 11501356 TI - Long-term follow-up of patients' status after gastric bypass. AB - BACKGROUND: We report a long-term (13-15 year) follow-up of a cohort of 100 patients who underwent gastric bypass for morbid obesity. METHODS: Sources of information include baseline data collected before surgery and information obtained at follow-up interview including data on weight history, psychosocial functioning, and medical complications. RESULTS: Mean age at follow-up was 56.8 years. The mean weight loss at long-term follow-up was 29.5 kg (range -13.6 to 93.6 kg). Three subjects weighed more at long-term follow-up than before the operation. Overall, 74% of those interviewed indicated that the gastric bypass had benefited them in terms of their physical health. However, 68.8% reported continued problems with vomiting and 42.7% with "plugging". Eight had died. CONCLUSION: The findings in this study suggest that at long-term follow-up the majority of individuals who have undergone gastric bypass feel that the procedure benefited them, although some complications including difficulties with "plugging" and vomiting were present at long-term follow-up. PMID- 11501357 TI - A biodegradeable membrane from porcine intestinal submucosa to reinforce the gastrojejunostomy in laparoscopic Roux-en-Y gastric bypass: preliminary report. AB - BACKGROUND: A Silastic ring has been used to prevent dilation of the gastrojejunostomy in Roux-en-Y gastric bypass (RYGBP). The use of a bio-membrane may prevent dilation of the anastomosis without the risks associated with prostheses. The aim of this study was to evaluate the feasibility and safety of applying such a bio-membrane around the gastrojejunostomy in Laparoscopic RYGBP (LRYGBP). METHODS: We used a new bio-membrane, that is derived from porcine small intestinal submucosa (SIS) and acts as a scaffolding for the ingrowth of connective tissue. Over a 4-month period, 14 LRYGBP patients had their proximal anastomosis wrapped with 10 x 2.5 cm SIS by a single surgeon. We compared these patients to a control group of LRYGBP patients matched for BMI. RESULTS: The average age of the patients was 35.0 years (control group: 45.1 years). The patients had a mean initial BMI of 44.7 kg/m2 (+/- 5.9) standard error, and the control subjects had a mean initial BMI of 46.7 kg/m2 (+/- 6.5). SIS application took a mean time of 11 (+/- 3) minutes without any intraoperative complication. The median hospital stay was 3.5 days in the experimental group and 3.7 days in controls. Three patients developed a symptomatic stenosis at the gastrojejunostomy following surgery. In the control group there were two stenoses. At an average follow-up of 87 days (controls: 95 days), the mean reduction in BMI was 7.8 (+/- 0.8) kg/m2 [controls 8.6 kg/m2 (+/- 1.5)]. CONCLUSION: Application of SIS around the gastrojejunostomy in patients undergoing LRYGBP is feasible and safe. Further follow-up is required, however, to evaluate the effectiveness in preventing dilation of the anastomosis. PMID- 11501358 TI - Avoidance of weight regain after gastric bypass. PMID- 11501359 TI - Obesity-associated disorders before and after weight reduction by vertical banded gastroplasty in morbidly vs super obese individuals. AB - BACKGROUND: The amount of excess weight which must be lost in order to cure or to improve disorders associated with obesity remains unknown. This study was designed to compare super obese and morbidly obese patients in terms of weight reduction following VBG and to investigate the effects of postoperative weight changes to preexisting co-morbidities. METHODS: 125 patients underwent VBG. Group A consisted of 80 morbidly obese patients (64%) and group B consisted of 45 super obese patients (36%). Preoperative examination was planned to identify and determine the severity of any disorders associated with obesity, that the patients may have had. Following VBG, all patients were followed-up at regular time periods, for at least 2 and up to 4 years. The progress of preexisting co morbidities was evaluated and carefully recorded. RESULTS: Among the 80 patients of Group A, there were 240 total co-morbidities (3 per patient), and in group B there were 196 co-morbidities (4.35 per patient) preoperatively. Dyspnea during fatigue and arthritis were found at statistically higher incidence in the super obese category. At the end of the second postoperative year, greater weight loss in terms of number kilograms was seen in patients in group B, but these patients did not reach a BMI lower than 35, while patients in group A had mean BMI below 30. In group A, 66% of the co-morbidities completely resolved, 19% significantly diminished and 15% remain unchanged. In group B, the respective percentages were 53%, 27.5% and 19.5%. However, after weight reduction by VBG a significant number of co-morbidities remain in the super obese patients (92 or 2.044 per/patient), and this is believed to be due to the greater remaining excess weight. CONCLUSION: Reduction of body weight by VBG is associated with resolution or improvement of a significant number of the obesity-associated disorders. However, super obese patients remain obese after surgery, and this results in two-fold higher remaining morbidity. PMID- 11501360 TI - Vertical banded gastroplasty in the Aegean Region of Turkey. AB - BACKGROUND: Obesity is increasing in Turkey. The first experience with vertical banded gastroplasty (VBG) with regular intermediate-term follow-up in the Aegean Region of Turkey is presented. METHODS: From November 1993 to August 1999, 40 morbidity obese patients underwent VBG. The patients were evaluated on the basis of excess weight loss (EWL) and satisfaction with the operation. RESULTS: Regular follow-up was obtained in 38 patients (95%), with mean follow-up 27.2 months (14 85). Average preoperative body weight (BW) was 141.4 kg (93-238) and body mass index (BMI) was 52.3 kg/m2 (41-77.8). Average EWL was 64.1% (21.2-92.3). Average postoperative BW and BMI were 93.3 kg (70-145) and 34.4 kg/m2 (25.1-53) respectively. 35 of 38 patients (92%) lost more than 25% of EW and 28 of 38 (73.6%) lost more than 50% of EW. After weight loss, hypertension disappeared or improved in 86% of patients and diabetes resolved in 75%. Sleep apnea disappeared in 100% of patients. The early and late complication rates were 7.9% and 15.8% respectively. 33 of 38 patients (87%) were satisfied with the operation. CONCLUSION: VBG was safe and effective, resulted in acceptable weight loss, and the vast majority of patients were satisfied. PMID- 11501361 TI - Complications after gastroplasty and gastric bypass as a primary operation and as a reoperation. AB - BACKGROUND: Since 1991 we performed vertical banded gastroplasty (VBG) as our surgical treatment of choice for morbid obesity in 680 patients, and since 1996 we also performed Roux-en-Y gastric bypass (RYGB) in 36 patients. For revisional surgery, the surgeons chose procedures based on their experience. METHODS: We recorded early complications (0.6%) and late complications (8.5%) after the primary operations. When staple-line disruption or stenosis of the banded stoma occurred after VBG, revisional surgery was performed with re-VBG or conversion to RYGBP. Some early complications needed emergency operation for bleeding or gastric perforation. RESULTS: Mortality was zero. Reoperation with re-VBG and RYGBP was effective in all patients, but for many, a long stay in hospital was necessary because reoperation had a high rate of early and late complications, 33.8% and 21.8% respectively. CONCLUSION: The treatment of complications after VBG with re-VBG and RYGBP had danger. We believe that when VBG failure occurs, to avoid dangerous complications again, we should perform a biliopancreatic diversion, which does not involve a gastric restriction. PMID- 11501362 TI - Outcome of biliopancreatic diversion in subjects with Prader-Willi Syndrome. AB - BACKGROUND: In Prader-Willi Syndrome (PWS), mental retardation and compulsive hyperphagia cause early obesity, the co-morbidities of which lead to short life expectancy, with death usually occurring in their 20s. Long-term weight loss is mandatory to lengthen the survival; therefore, the lack of compliance in voluntary food restriction requires a surgical malabsorptive approach. METHODS: 15 PWS subjects were submitted to biliopancreatic diversion (BPD) and followed (100%) for a mean period of 8.5 (4-13) years. BPD consists of a distal gastrectomy with a long Roux-en-Y reconstruction which, by delaying the meeting between food and biliopancreatic juices, causes an intestinal malabsorption. Indication for BPD was BMI > 40 or > 35 with metabolic complications. Preoperative mean age was 21 +/- 5 years, mean weight 127 +/- 26 kg, and mean Body Mass Index (BMI, kg/m2) 53 +/- 10. According to Holm's criteria, all of the subjects had a total score > or = 8. IQ assessment was performed in each subject, with a mean score of 72 +/- 10. An arbitrary lifestyle score was given to each subject. RESULTS: No perioperative complications were observed. Percent excess weight loss (%EWL) was 59 +/- 15 at 2 years and 56 +/- 16 at 3 years, and then progressive regain occurred; at 5 years %EWL was 46 +/- 22 and at 10 years 40 +/- 27. Spearman rank test failed to demonstrate any correlation between weight loss at 5 years and patient data, except with lifestyle score (Spearman r = 0.8548, p < .0001). Current mean age is 31 +/- 7 years. CONCLUSION: BPD has to be considered for its value in prolonging and qualitatively improving the PWS patient's life. PMID- 11501363 TI - Total intravenous anesthesia versus single breath technique and anesthesia maintenance with sevoflurane for bariatric operations. AB - BACKGROUND: The choice of anesthetic technique for general anesthesia in morbidly obese patients remains controversial. We aimed to compare blood gases, recovery and hemodynamic parameters using TIVA and sevoflurane anesthesia in bariatric surgery. METHODS: The study was performed with permission of the ethics committee. We studied 40 morbidly obese patients allocated to 2 groups. The total i.v. anesthesia (TIVA) group was named Group T, and the sevoflurane group was named Group S. In Group T, anesthesia induction was achieved with propofol. In Group S, anesthesia induction was achieved by sevoflurane with single breath technique, with maintenance provided with 1-2% volume sevoflurane. Student t, Chi square and ANOVA tests were used for data analysis; p-value < 0.05 was considered statistically significant. RESULTS: There was no significant difference between the 2 groups in demographic data, blood gas values and recovery characteristic. Hemodynamic values were significantly lower in Group T than Group S, during and after the operative period. CONCLUSION: While sevoflurane induction and maintenance is a suitable anesthetic modality for obese patients, TIVA can be applied easily in those patients possessing no extra risk factors other than morbid obesity. PMID- 11501364 TI - Improved scintigraphic assessment of occult leakages in adjustable gastric bands using 99mTc-labelled human albumin colloid. AB - BACKGROUND: Adjustable gastric banding has become the preferred procedure in Europe for the treatment of morbid obesity. A complication of this treatment is the presence of leakage in the system. The knowledge of the localization of the leak is essential for planning the reoperation procedure. PATIENTS AND METHODS: In a series of 325 adjustable gastric bandings, we observed 10 band leakages. In 3 cases, fluoroscopy failed to detect fluid extravasation. We present a sensitive and simple scintigraphic method for the detection of gastric band leakage in these patients using 37 MBq of a 99mTc-human albumin colloid suspension. Imaging was started immediately after the dose application. During the first minute, images were acquired dynamically with 1 frame every 2 seconds. Afterwards, a static image was obtained every 10 minutes, up to 60 minutes post-injection. Then the system was emptied completely to detect tracer extravasation and consecutive reabsorption during 60 minutes. The study was analyzed by using the regions of interest (ROI) technique drawn on the following points: injection reservoir, tube, anterior band, posterior band, and an adjacent region which was taken as background. RESULTS: During the first hour, ROI analysis showed a clear diminution in the number of counts contained in the defect parts of the band, whereas it remained constant in the other locations. After system emptying, detection of tracer extravasation further strengthened the diagnosis of band leakage in all patients. CONCLUSION: This new approach using the 99mTc-human albumin colloid suspension with ROI analysis is an effective and simple method to detect occult leakages in adjustable gastric bands which escaped detection by fluoroscopy. In contrast to previously described scintigraphic methods, this investigation is able to demonstrate the exact site of leakage. PMID- 11501365 TI - Laparoscopic band repositioning for pouch dilatation/slippage after gastric banding: disappointing results. AB - BACKGROUND: Pouch dilatation with or without slippage of the band is a serious complication of gastric banding, often attributed to initial malpositioning of the band. Food intake is increased, and weight regain occurs. Progressive rotation of the band follows, leading to functional stenosis and dysphagia. Reoperation is necessary in most cases, and may consist of band removal, band change, band repositioning, or conversion to another bariatric procedure. MATERIAL AND METHODS: The study consisted of chart review of all patients who underwent laparoscopic repositioning of the band for pouch dilatation/slippage, and long-term follow-up through regular office visits and phone calls. RESULTS: Among 272 patients who had laparoscopic gastric banding, 20 (7.3%) developed pouch dilatation and/or slippage, of whom 19 underwent reoperation. Laparoscopic band repositioning was performed in 9 patients. One of them developed an intraabdominal collection postoperatively and required percutaneous CT-guided drainage. Recovery was uneventful in the other 8. Follow-up since reoperation varies from 13 to 42 months (mean 20 months). The result was good in 2 patients who lost further weight, satisfactory in 1 whose weight remained stable, and unsatisfactory in 6 patients. Weight loss was insufficient in 2, dilatation recurred in 2, and band infection or erosion developed each in 1 patient. 5 patients required further surgery: band removal in 3 and conversion to gastric bypass in 2. CONCLUSIONS: Laparoscopic band repositioning is feasible and safe if pouch dilatation and/or slippage develops after gastric banding. The mid-term results are disappointing in two-thirds of the patients. In some patients, pouch dilatation could result from poor adjustment to diet restriction rather than merely from original malplacement. Conversion to gastric bypass may be a better option in these cases. PMID- 11501366 TI - Pseudo-achalasia following laparoscopically placed adjustable gastric banding. AB - BACKGROUND: The complication of pseudo-achalasia may occur after laparoscopic adjustable gastric banding (LAGB) in patients with normal band position and normal stomal width. We hypothesized that this complication occurs especially in patients with preexisting insufficiency of the lower esophageal sphincter (LES), who show poor compliance secondary to lacking the sensation of satiety and who therefore also have insufficient weight loss at follow-up. METHODS: Early and late postoperative barium meal studies of 120 LAGB patients were retrospectively analyzed to identify patients who developed esophageal widening and dysmotility despite normal band position and normal stomal width. Results were compared with preoperative endoscopies, clinical findings, each patient's compliance with dietary instructions and postoperative weight loss. RESULTS: 9/120 patients developed pouch dilatation, esophageal widening and esophageal dysmotility as a late complication, despite normal band position and normal stomal width. All these patients had shown preexisting insufficiency of their LES endoscopically. They all showed bad compliance with dietary instruction, and they all abused their distal esophagus as an additional pouch. 7 of these patients presented with insufficient weight loss at follow-up, whereas of 3 other patients with pre existing LES insufficiency who had shown good compliance, only 1 showed insufficient weight loss. Insufficient weight loss after 1 year was significantly more common in patients with pre-existing LES insufficiency (8/12 patients, 67%) than in patients with a competent LES (26/108 patients, 24%). CONCLUSION: Patients with pre-existing LES insufficiency appear to be at risk for pouch dilatation and esophageal decompensation despite normal band position and normal stomal width. These patients are prone to show lack of satiety and poor compliance with dietary instruction, use of their lower esophagus as additional space for food, and tend to have insufficient weight loss. Preoperative manometry should be used to identify such patients, where the indication for gastric banding should be discussed very critically. PMID- 11501367 TI - Weight reduction by means of intragastric device: experience with the bioenterics intragastric balloon. AB - BACKGROUND: A new intragastric balloon is available for weight reduction. METHODS: Patients consulting for a weight reduction plan and refusing any kind of actual surgery, or suffering from obesity but not meeting the IFSO standards for surgery, were offered the possibility of weight reduction by the BioEnterics intragastric balloon (BIB). A preoperative questionnaire was completed by all patients, inquiring about medical history, co-morbidity factors, dietary habits, previous treatments for weight and social, psychological, relational and economic impact of the obesity. BIB placement was done on an inpatient basis, under general anesthesia. Inflation was standardized at 500 ml saline. Intravenous antiemetic and spasmolytic drugs were given to control post-insertion nausea for 24 hours, and oral medication was administered on the patient's discharge. A standard 800 calorie diet was prescribed after dietitian's consultation. Extraction of the balloon was left to the patient's discretion at 3 or maximum 6 months after placement. Patients choosing for the maximal period received a formal invitation to extract the balloon. Evaluation of weight reduction was done at extraction and by questionnaire. RESULTS: 126 patients (5 M, 121 F) with mean age of 35.6 years (20-62) were included after preoperative evaluation. Mean preoperative BMI was 37.7 kg/m2 (26.7-57.7 kg/m2), with a mean initial excessive weight of 35.3 kg (8.8-96.4 kg) and mean initial % excess weight of 32.2 (6.3 102). 69 patients were eligible for review; mean excess weight loss after 3 months was 48.6% and after 6 months 50.8%. Mean weight loss was 15.4 kg (0-35 kg). 76.8% of the patients (41/69) complained of severe nausea and vomiting lasting an average of 1 week (1 day-6 months), resulting in 3 patients in early removal of the balloon (at 1 day, 1 week, 1 month after placement respectively). 2 patients suffered gastric perforation presenting as acute peritonitis 3 and 4 months after placement and were operated. Extraction of the balloon was performed in 3 patients after 3 months and in 66 patients after 6 months. In 11 patients (22%), esophagitis was present (8 grade 1, 2 grade II, 1 grade III), and one patient showed diffuse gastric erosion. One patient required removal of the balloon by rigid esphagoscopy following technical failure of the endoscopic extraction device. 45 patients replied to the mailed, questionnaire; 15% (7/45) were very satisfied, 13% (6/45) satisfied, 22% (10/45) reasonably satisfied, 8.8% (4/45) unsatisfied and 40% very unsatisfied. Degree of satisfaction correlated poorly with weight loss. Results may be better with close continuous guidance by a counselor. CONCLUSION: BIB as a means of weight reduction in the obese patient led to a 50.8% loss of excess weight after 6 months. Although severe morbidity can occur, the BIB provides a means for short-term weight reduction in conjunction with dietary measures. PMID- 11501368 TI - The BioEnterics Intragastric Balloon (BIB): how to use it. AB - BACKGROUND: The authors have been using the new saline-filled BioEnterics intragastric balloon (BIB) since 1995. METHODS: By now, more than 650 BIBs have been implanted on our Unit. RESULTS: Because of close collaboration between the engineers and the users, and after several modifications, an easy-to-use balloon is available. In addition, we have developed 3 instruments which provide easy and quick removal of the BIB. CONCLUSION: The device, supported by a competent motivated team, is another modality available for weight loss. PMID- 11501369 TI - Laparoscopic inflatable band with Roux-en-Y gastric bypass. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGBP) leaves a large blind gastric segment, which is inaccessible for conventional endoscopy. METHOD: A case is reported, describing a variation of laparoscopic RYGBP by partitioning the stomach by an inflatable band rather than by stapling or division. RESULTS: The stomach was partitioned into a proximal 15 cc pouch and a distal part by an adjustable gastric band. A RYGBP was fashioned from the proximal pouch. 9 patients were treated with this technique: 7 as an initial procedure and 2 after previous gastric banding which had been followed by insufficient weight loss. 1 of these latter patients developed erosion of the band through the gastrojejunostomy 7 months postoperatively. CONCLUSION: Laparoscopic proximal RYGBP with inflatable-band gastric partitioning is feasible. Erosion of the band though the gastrojejunostomy, however, might be a serious side-effect of this technique. PMID- 11501370 TI - The inflatable band and exercise machines. PMID- 11501371 TI - More on mini-gastric bypass. PMID- 11501372 TI - Post-transcriptional light regulation of nuclear-encoded genes. AB - A significant number of studies have detected a post-transcriptional component in the light responses of nuclear genes. As yet there are few in-depth studies of the mechanism(s) involved, and it seems likely some additional examples have been missed. For instance, transcriptional responses have sometimes been inferred on the basis of experiments with translational fusions containing both the promoter and 5' UTR of the test gene, but we now know that elements within the 5' UTR can mediate post-transcriptional light responses. Similarly, because of possible changes in translation rates and protein turnover, the common assumption that mRNA levels directly dictate protein levels is tenuous at best. It is no longer permissible to assume that the biological effect of a gene is a simple function of its transcription. Thus it is likely that with careful experimental design, reports of nuclear-encoded post-transcriptional gene regulation will become increasingly prevalent. PMID- 11501373 TI - Novel methods of introducing pest and disease resistance to crop plants. PMID- 11501374 TI - Elucidation of biosynthetic pathways by retrodictive/predictive comparison of isotopomer patterns determined by NMR spectroscopy. PMID- 11501375 TI - Are gene silencing mutants good tools for reliable transgene expression or reliable silencing of endogenous genes in plants? PMID- 11501376 TI - Manipulating plant viral RNA transcription signals. PMID- 11501377 TI - Genetic engineering strategies for hematologic malignancies. PMID- 11501378 TI - Telomerase and cancer. AB - The past few years have brought a flood of new information to the telomerase field. The identification of multiple components of both the telomere and telomerase, the understanding of the importance of telomere maintenance to the long term viability of cells, and the demonstration of the utility of telomerase inhibition in limiting tumor cell growth all convene to provide great enthusiasm for the prospects of targeting the telomerase enzyme in cancer. However, there is clearly much to be learned. Because tumor cells evolve under powerful selection, the emergence of non-telomerase based mechanisms for telomere maintenance should be examined closely. Additionally, the nature of telomerase regulation is currently only poorly understood. More work on the tumor specific regulation of telomerase activity might provide either more opportunities for telomerase inhibition, or more skepticism, as a tumor cell might possess mechanisms for upregulating telomerase activity in the presence of inhibitors. The potential for such regulation has already been observed in certain cell types (46). Currently, the field is intensively investigating the biology and applications of telomere and telomerase biology. In it are great hopes that these fundamental cellular processes might be manipulated to success in the treatment of cancer. PMID- 11501379 TI - Targeted gene repair in mammalian cells using chimeric oligonucleotides. PMID- 11501380 TI - Exploring the mechanism of action of insecticidal proteins by genetic engineering methods. PMID- 11501381 TI - Enzyme stabilization by directed evolution. PMID- 11501382 TI - ET-cloning: think recombination first. PMID- 11501383 TI - Growth and genetic modification of human beta-cells and beta-cell precursors. PMID- 11501384 TI - Open surgery has not had its day. PMID- 11501385 TI - Arterial reconstruction for lower limb ischemia. AB - The treatment of advanced atherosclerosis involving the lower extremities has undergone considerable evolution over the last several decades. Current strategies hinge on an appreciation of the natural history of disease, the overall health status of the patient, and an armamentarium of endovascular and open surgical reconstructive techniques that may be tailored to optimize outcome for the individual patient. Patients with aorto-iliac disease have a variety of available options with generally good results. For infrainguinal disease, surgical bypass using autogenous vein is the mainstay of interventional therapy and will remain so for the foreseeable future. Increasing medical and surgical challenges are presented by this population, particularly as aggressive medical treatment of risk factors leads to an ongoing decline in cardiovascular mortality, combined with a dramatic increase in the prevalence of diabetes. The future, which is now already at hand, will bring the application of genomics, with a potential for altering the progression of disease as well as extending the long-term benefits of reconstruction. PMID- 11501386 TI - Complications of thrombolytic therapy for peripheral arterial occlusions. PMID- 11501387 TI - Amputations in diabetic patients: a plea for footsparing surgery. AB - The authors observed a rather high rate of primary major amputation (above-knee or below-knee) performed for diabetic foot problems as well as an important revision rate for minor amputations (forefoot or toe) in diabetics. They reviewed their experience in order to compare it with more recent data from the literature, pleading for foot-sparing surgery. From 1993 to 1998, 186 amputations were performed on 146 diabetic patients. The cause of foot ulcers was neuropathy in 43 of them (51 episodes of diabetic foot problems) while in the remaining 103 patients (135 episodes of diabetic foot problems), diabetic macroangiopathy (absent ankle pulses) was on cause. For neuropathic foot problems, amputations were almost minor, resulting in a limb salvage rate of 90%. Only five of these patients (12%) had primary major limb amputation versus 43 of the dysvascular patients (42%). The reasons for major amputation by first intention were extensive tissue loss, intractable infection or non-reconstructible occlusive vessel disease, as judged by the surgeon. A foot-sparing surgery was attempted in 92 dysvascular cases. In only 44 of them, a preliminary vascular repair was performed. Twenty eight percent of the primary toe amputations and 24% of the forefoot amputations required secondary revision to a more proximal level. Minor amputations in case of diabetic neuropathy were characterized by a more favourable outcome: only 14% of the toe and 9% of the forefoot amputations failed. During follow-up, only 63% of the major amputations regained an autonomic walking capability with their prosthesis. Wound healing problems in diabetic foot are mainly due to infection and poor tissue perfusion. An aggressive control of the infection and distal revascularization of calf- or foot arteries, whenever possible, could improve the results of diabetic foot surgery. The poor functional recovery after major amputation (only 63% autonomic gait with limb prosthesis) argues for foot-sparing surgery whenever possible. PMID- 11501388 TI - Beating heart surgery using the Octopus tissue stabilizers: initial experience including triple vessel disease and high-risk patients. AB - OBJECTIVE: The aim of this study was to evaluate initial results in beating heart coronary artery bypass grafting performed on patients with multivessel disease, using suction-based tissue stabilizers. METHODS: Forty patients (mean age 65.5 years, range 39-83) underwent beating heart coronary bypass by median sternotomy, with the use of the Octopus 1 (n = 27) and Octopus 2 (n = 13) devices. Twenty patients had unstable angina, four had pulmonary oedema, and four required preoperative intra-aortic balloon counterpulsation. Five interventions were repeat procedures. Mean ejection fraction was 54.4%(range 20-82%). Eighteen patients had triple-vessel disease, 18 patients had double-vessel disease and > 50% left main stem stenosis was present in six patients (isolated or in association). The average number of distal anastomoses was 2.5 (range 1-5, total 101). A mean of 1.5 (range 1-3) anastomoses was achieved with arterial grafts (45 mammary and 12 gastroepiploic arteries). RESULTS: Immediate graft patency was evaluated by Doppler flowmeter and five anastomoses were successfully corrected, based on an occlusion pattern. The perioperative myocardial infarction rate was zero. Transient episodes of supraventricular arrhythmias were detected in 19 patients. The administration of dobutamine at an inotropic concentration > 5 gamma-1 kg-1 min-1 was required in one patient. There were two in-hospital deaths (one non-cardiac-related). Overall survival and cardiac event-free rate at 20 months were 92.4% +/- 4.2% and 81.8% +/- 11.6%, respectively. CONCLUSIONS: Satisfactory results can be achieved in multivessel disease high-risk patients with beating heart coronary bypass surgery. PMID- 11501389 TI - Laparoscopic inguinal hernia repair: review of 6 years experience. AB - Since 6 years, the totally extraperitoneal laparoscopic hernia repair has become our procedure of choice to manage inguinal hernia in adult patients, especially for bilateral hernias and recurrences after classical anterior repair. Between March 1993 and March 1999, 976 patients underwent 1259 hernia repairs by an endoscopic total extraperitoneal approach. A large polypropylene prosthesis (15 x 15 cm) is placed and covers all potential defects. Follow-up on patients ranged from 6 to 79 months (mean, 39 months). Per- and postoperative morbidity and complications were acceptable (8.4%) and included conversion to open surgery (0.4%), bleedings (0.3%), urinary retention (4.2%), seromas (2.7%), neuralgias (0.2%), vague persistent groin discomfort (0.4%), orchitis (0.08%) and sigmoido cutaneous fistula (0.08%). Recurrence rate so far is 0.1%. This retrospective study shows that the totally extraperitoneal repair for inguinal hernia should have a promising future because of low morbidity and low recurrence rate. PMID- 11501390 TI - Multiple postoperative thromboembolic events and fatal pulmonary embolism: the white clot syndrome. A case report. PMID- 11501392 TI - Gathering evidence-based information from the Internet. PMID- 11501391 TI - Congenital right-sided diaphragmatic hernia in an elderly patient. A case report. AB - Congenital diaphragmatic hernia is a rare condition in adulthood. It is even more exceptional when located on the right side. We describe a case of right-sided congenital diaphragmatic hernia in a 74-year old woman. The diagnosis was only made when the patient developed an acute intestinal obstruction after a laparotomy for a gynaecological benign tumour. The treatment of this condition is discussed. In our opinion, in elderly, the advantages of an elective operation in asymptomatic congenital diaphragmatic hernia have to outweigh the risks of the operation. However, if the patient shows symptoms of pulmonary dysfunction or motility disorders of the gut, or even when he has to undergo a laparotomy for a different reason, we advise to repair the hernia in order to prevent complications. PMID- 11501393 TI - Evidence-based medicine, cost-containment and necessary choices. PMID- 11501394 TI - Evidence-based medicine, cost-containment, care effectiveness: is it a new trilogy aimed at transforming the surgical mystique or the reality of double standards? AB - Clinically relevant attitudes and guidelines issued by a rational Evidence-Based Medicine (EBM) approach integrate individual clinical expertise with the best available external clinical evidence from systematic research. However, many surgeons, while considering the ultraliberal world they are practicing in and fearing that the primary goal of managed care in a market environment is reducing cost in order to make profit or decrease spending, remain suspicious of this kind of tentative protocol-driven medicine when applied to surgical practice. If surgeons want to develop a health policy agenda that emphasizes patient care issues above provider's or payor's interests, they should also enhance education programs, improve continuing objective assessment of the way surgery is performed, face moral issues raised by innovation, and assume an increased leadership role in sound critical evaluation of nonvalidated new techniques. They should no more consider EBM as a weapon turned against the surgical profession, but as a tool that may provide some answers to chronically unresolved questions in the evolving art of surgery. PMID- 11501395 TI - Isoquinoline alkaloids from Mahonia aquifolium stem bark are active against Malassezia spp. AB - Bioassay directed fractionation of crude extract from Mahonia aquifolium led to the isolation of fraction A (bisbenzylisoquinoline alkaloid complex, BBI) and a fraction of protoberberine alkaloids, where the major compounds berberine and jatrorrhizine were isolated as their iodides. The antifungal activity of the crude extract, two protoberberine alkaloids and BBI from M. aquifolium stem bark were evaluated against six strains of Malassezia spp. The compounds tested were generally found to possess only weak to moderate antifungal properties: the MICs for individual strains were in the range < or = 50-> or = 1000 mg/L. PMID- 11501396 TI - Antibacterial and mutagenic activities of new isothiocyanate derivatives. AB - Nine newly synthesized isothiocyanate derivatives were demonstrated to posses antibacterial and genotoxic activities in vitro. 4-Hydroxybutyl isothiocyanate exhibited a broad antibacterial effect, with MIC values of 762 mumol/L for Staphylococcus aureus and Escherichia coli. Ethyl 4-methylsulfoxidobutanoate had the highest antibacterial activity in Gram-positive bacteria, the MIC value being 425 mumol/L for S. aureus. The highest tested concentrations of ethyl 4 isothiocyanatobutanoate and 4-hydroxybutyl isothiocyanate produced a bacteriocidal effect in Gram-positive bacteria. The compounds showed no mutagenic effects on Salmonella typhimurium tester strains TA 98 and TA 100, either in the absence or in the presence of a metabolically active microsomal S9 fraction from rat liver using standard Ames test. PMID- 11501397 TI - The restriction-modification system in Streptomyces flavopersicus. AB - To clone bifunctional vectors in streptomycetes, it was necessary to define the restriction-modification system of Streptomyces flavopersicus. Plasmid DNA from bifunctional vectors pIJ699 and pXED3-13, isolated from E. coli strains with different methylation systems: E. coli DH5 alpha (dam+ dcm+), E. coli MB5386 (dam dcm), E. coli CB51 (dam dcm+), E. coli NM544 (dam+ dcm), was used for transformation of protoplasts from strain S. flavopersicus. Restriction of dcm methylated DNA from S. flavopersicus was established. As a host in the intermediate cloning strain E. coli NM544 (dam+ dcm) should be used, as the dcm transmethylase-dependent strain S. flavopersicus does not process DNA from this strain. PMID- 11501398 TI - Relationship between production of 3-indoleacetic acid and peroxidase-laccase activities depending on the culture periods in Funalia trogii (Trametes trogii). AB - The relationship between production of 3-indoleacetic acid (IAA) and peroxidase and laccase activity was investigated in white-rot fungus Funalia trogii (Trametes trogii). F. trogii produced IAA and peroxidase and laccase as both primary metabolite and secondary metabolite; the levels of IAA may be influenced by peroxidase and laccase. A correlation exists between the levels of IAA and peroxidase-laccase activity. PMID- 11501399 TI - Penicillin V production by Penicillium chrysogenum in the presence of Fe3+ and in low-iron culture medium. AB - Late-exponential-phase Penicillium chrysogenum mycelia grown in a complex medium possessed an intracellular iron concentration of 650 mumol/L (2.2 +/- 0.6 mumol per g mycelial dry mass). This iron reserve was sufficient to ensure growth and antibiotic production after transferring mycelia into a defined low-iron minimal medium. Although the addition of Fe3+ to the Fe-limited cultures increased significantly the intracellular iron levels the surplus iron did not influence the production of penicillin V. Supplements of purified major P. chrysogenum siderophores (coprogen and ferrichrome) into the fermentation media did not affect the beta-lactam production and intracellular iron level. Neither 150 nor 300 mumol/L extracellular Fe3+ concentrations disturbed the glutathione metabolism of the fungus, and increased the oxidative stress caused by 700 mmol/L H2O2. Nevertheless, when iron was applied in the FeII oxidation state the oxidative cell injuries caused by the peroxide were significantly enhanced. PMID- 11501400 TI - Industrial and agricultural wastes as substrates for laccase production by white rot fungi. AB - White-rot fungi, Coriolus versicolor and Funalia trogii, produced laccase on media with diluted olive-oil mill wastewater and vinasse. Addition of spent cotton stalks enhanced the laccase activity with a maximum after 12 d of cultivation. PMID- 11501401 TI - Individual and combined effects of berberine and santonin on spore germination of some fungi. AB - Berberine and santonin were isolated from rhizomes of Berberis aristata and unexpanded flower buds of Artemisia maritima, respectively. Efficacy of these two chemicals individually as well as of their mixtures, was tested against spore germination of some saprophytic and obligate fungi. While berberine individually was effective against most of the fungi, Helminthosporium spp. were least affected even at the highest dose (1500 ppm). Santonin was equally effective against several fungi. Mixture of both alkaloids found to be more effective than individual ones. Keeping the dose of berberine constant and santonin at two different concentrations (viz. 250 and 500 ppm) the spore germination of Helminthosporium oryzae was stimulated. Increasing concentration of santonin inhibited the spore germination of all other fungi tested, Colletotrichum capsici being affected only by 20 and 5% (at berberine concentration of 250 and 500 ppm, respectively). On the other hand, santonin being constant and berberine at different concentrations, the mixture was effective against all the fungi. PMID- 11501402 TI - Decolorization of Orange II dye by white-rot fungi. AB - Agitation, temperature, inoculum size, initial pH and pH of buffered medium affected the decolorization of Orange II dye by Coriolus versicolor and Funalia trogii. The optimum temperature and initial pH value for decolorization were 30 degrees C and 6.5-7.0, respectively; pH 4.5 was the most efficient in buffered cultures. High decolorization extents were reached at all agitation rates. At an inoculum size of more than 1 mL, the extent of decolorization changed only slightly. High extents were obtained using immobilized fungi at repeated batch mode. PMID- 11501403 TI - Yeast-like microorganisms in eye infections. AB - The proportion of yeast species involved in eye infections in 11 patients was examined. The presence of yeast organisms as causative agents of endophthalmitis was found in corneal smears (n = 4), conjunctival swabs (4), and vitreous fluid (3). Altogether 5 strains of Candida albicans, 2 strains of C. krusei and one strain each of C. guilliermondii, C. parapsilosis, C. tropicalis and Cryptococcus neoformans were isolated from the clinical material. The hematogenic origin of endophthalmitis was proved in 7 cases on the basis of positive blood samples and in 2 cases by the isolation of yeasts from the tip of an intravenous catheter. Endophthalmitis-supporting risk factors such as indwelling intravenous catheters, prolonged use of broad-spectrum antibiotics and chemotherapy, surgical intervention, diabetes mellitus, and malignancy were observed in the patients. PMID- 11501404 TI - Occurrence of variants with temperature-dependent susceptibility (TDS) to antibiotics among Stenotrophomonas maltophilia clinical strains. AB - Susceptibility to 20 antibiotics was tested in 104 Stenotrophomonas maltophilia strains at 37 and 30 degrees C by means of a dilution micromethod to verify the phenomenon of temperature-dependent susceptibility (TDS). Trimethoprim sulfamethoxazole, pefloxacin and ofloxacin were the most active preparations at 37 degrees C (93, 90, and 86% of susceptible strains, respectively), whilst trimethoprim-sulfamethoxazole, cefoperazone-sulbactam and pefloxacin performed best at 30 degrees C (94, 94, and 76% of susceptible strains, respectively). Variants 37TDS (minimum inhibitory concentration, MIC, of tested antibiotics at least 4-times lower at 37 than at 30 degrees C) occurred in 60%. Variants 30TDS (at least 4-times lower value of MIC at 30 than at 37 degrees C) were found in 7.7%. Both variants in susceptibility to tested antibiotics appeared in 23.1%, whilst neither of them was observed in 9.6%. The 37TDS phenomenon was recorded most of all with gentamicin (51% of strains), amikacin (47), colistin (44) and tetracycline (34). The 30TDS phenomenon was found particularly with cefoperazone sulbactam (16.0% of strains) and colistin (10.0%). The above phenomena may be due to changes in membrane permeability, temperature-dependent ribosomal changes, and insufficient adaptation to higher temperatures of some strains of the originally environmental species S. maltophilia. PMID- 11501405 TI - Effect of enterocin CCM 4231 on Listeria monocytogenes in Saint-Paulin cheese. AB - The bacteriocin production by Enterococcus faecium strain in cheese milk and cheese was demonstrated. Purified enterocin CCM 4231 exhibited an anti-listerial effect during Saint-Paulin cheese manufacture. During cheese production the strain grew to a final concentration of 10.1 +/- 0.01 log CFU per mL per g in cheese. Then only a slight decrease of the cell concentration was noticed during ripening and was almost stable for 8 weeks. No significant differences in pH were observed between the experimental and reference cheeses. Bacteriocin production during cheese manufacture was detected only in milk samples and curd, reaching a level of 100 AU/mL. After addition of purified enterocin CCM 4231 (concentration 3200 AU/mL) into the experimental cheese, the initial concentration of 6.7 +/- 0.06 log CFU per mL of Listeria monocytogenes Ohio was reduced up to 1.9 +/- 0.01 log CFU per mL per g. After 6 weeks and at the end of the experiment the difference of surviving cells of L. monocytogenes Ohio in ECH was only one or 0.7 log cycle compared to the control cheese. Although enterocin CCM 4231 partially inhibited L. monocytogenes in Saint-Paulin cheese manufacture, an inhibitory effect of enterocin added was shown in 1-week cheese; however, it was not possible to detect bacteriocin activity by the agar spot test. The traditional fermentation and ripening process was not disturbed, resulting in acceptable end products, including sensory aspects. PMID- 11501406 TI - Candidaemia in cancer patients and in children in a neonatal intensive care unit. AB - The species most frequently found in the group of neonates was C. parapsilosis, its incidence reaching 48%. Other yeasts isolated from blood were C. albicans (41%), C. tropicalis (7%), C. krusei and C. pseudotropicalis (2% each). Six yeast species (C. albicans 44%, C. parapsilosis 28%, C. tropicalis 11%, C. krusei 8%, C. guilliermondii 6%, C. lusitaniae 3%) were detected in cancer patients. The presence of an intravenous catheter as a possible risk in the development of candidaemia was identified in all neonates and in 69% of cancer patients. C. krusei candidaemia was associated with prophylactic fluconazole therapy. PMID- 11501407 TI - Protein profile and biochemical properties of Bacillus circulans isolated from intestines of small free-living animals in Poland. AB - Forty-seven strains of Bacillus circulans isolated from the intestinal tract of free-living small mammals from the Narvia and Biebrza National Park (NE Poland) were compared with the electrophoretic whole-cell protein patterns on the basis of SDS-PAGE and biochemical characteristic using API tests (50 CHB and 20E). The strains were grouped into two clusters (I and II) at the similarity of protein pattern of 78% using the simple matching coefficient and clustering on unweighted pair group arithmetic average algorithm (SSM/UPGMA). Each of the clusters comprised reference strains. The rods included in the cluster I are characterized by higher biochemical activity in comparison with the bacilli II. PMID- 11501408 TI - NK cytotoxic activity and relative distribution of NK cells in 3-acetylpyridine influenced mice. AB - NK-cell cytotoxic activity and their relative distributions were studied in the spleen of female Lurcher mice with spontaneous olivopontocerebellar degeneration (C3H) and female athymic nu/nu mice (BALB/c) influenced by 3-acetylpyridine (the neurotoxin causing selective degeneration of cerebellar and inferior olive neurons in some rodent species). The congenital olivopontocerebellar degeneration in Lurcher mice is followed by only an insignificant increase of NK-cell cytotoxic activity (1.2 times). On the other hand, the congenital thymic dysgenesis in nu/nu mice is compensated by a substantial increase in cytotoxic activity (19.4-fold). The administration of 3-acetylpyridine (including prevalent neuronal destruction particularly in Lurcher mutants) caused a decrease of NK cell cytotoxic activities in all groups of mice (in Lurcher and C3H controls to 60 and 50%, respectively, and in nu/nu and BALB/c controls to 25 and 60%). Relative distributions of NK-cells in spleens of non-influenced and influenced animals were not significantly changed. Some fundamental immune mechanisms, such as the NK-cell cytotoxic activity, were demonstrated to be controlled by congenitally determined or artificially induced changes in both the nervous and the immune systems. PMID- 11501409 TI - Phylogenetic analysis of the rplA genes encoding ribosomal protein L1. AB - Previously we have identified the rplA gene encoding ribosomal protein L1 in Streptomyces aureofaciens. Sequence comparison of ribosomal protein L1 among several bacterial genera revealed a high level of conservation. Based on this conservation, these proteins were used as a phylogenetic tool to compare evolutionary relationships among eubacteria and archaebacteria. This phylogenetic analysis of L1 ribosomal proteins including the S. aureofaciens rplA gene product revealed, except similar bacterial groupings, some new evolutionary relationships. PMID- 11501410 TI - Ovine scrapie: priorities and importance. AB - Ovine and caprine scrapie occupies a unique place among animal transmissive spongiform encephalopathies (TSE). It is an object of intensive biomedicinal, ecological and economical studies. Its causative agents are demonstrably associated with the development of TSE in farmed minks, goats and moufflons. Ovine strains of scrapie occurring in North America (particularly in the USA) differ from strains which occur in Europe and were present at the onset of development of TSE in three species of deer living in free nature and in captivity in the USA. The studies dealing with the development of bovine spongiform encephalopathy (BSE) of the English type have indicated justifiably that its origin is associated with one (or more) heretofore unidentified ovine strain. The development of a variant form, the Creutzfeldt-Jacob disease in humans, and transmission of the BSE agent to several families of bovidae, felidae and primates, puts stress on its zoonotic potential. All this leads to the conclusion that domesticated sheep are the decisive reservoir species of animal TSE. They have been infected to an unknown extent with the causative agent of BSE probably through contaminated meat-bone meal. The occurrence of natural ovine prion isolates with properties similar to those of the BSE agent requires that scrapie should be included in the surveillance of human and animal TSE. At present, scrapie is a noticeable disease also in other than European Communities Member States. It is on the list B of the International Epizootics Office. Many countries have initiated control of ovine scrapie. It should therefore become a topical question also in Central and Eastern European countries. Elimination or even eradication of ovine scrapie (or its causative agents) from populations of small and large domestic ruminants is the prerequisite for prevention of penetration of ovine pathogenic prions into the human feed chain. Moreover, it should be ensured that these species will be able to produce foods of a new type (immunotrition and similar) or proteins with therapeutic effects in the near future. Our study established that the PrP genotype of Valachian rams, the Slovak autochthonous breed, contains also VRQ and ARQ alleles encoding the susceptibility to scrapie. Their selection is part of the improvement of Slovak Valachian sheep towards resistance to scrapie. PMID- 11501411 TI - Effects of subinhibitory concentrations of nitroxoline on the surface properties of Escherichia coli. AB - Nitroxoline (5-nitro-8-quinolinol; NIQ) at subinhibitory concentrations (sub-MIC) decreased the adherence of uropathogenic Escherichia coli to catheter surface and significantly enhanced cell surface hydrophobicity. The surface hydrophobicity increased in the presence of sub-MIC of NIQ and also in an excess of Mg2+. The effect of NIQ on the cell surface was not related to the bacteriostatic effect of this agent. The increase in nitrogen and decrease in phosphate content in the cell surface was found in the presence of NIQ. NIQ did not inhibit the expression of fimbriae. PMID- 11501412 TI - Dihexyl sulfosuccinate biodegradation by mixed cultures. AB - The primary biodegradation of dihexyl sulfosuccinate has been demonstrated using two single cultures and three mixed bacterial cultures of Comamonas terrigena. Primary biodegradation was found in all variants. The highest biotransformation rate and efficiency was observed in one of the single strains. No synergistic effect on degradation was found in mixed cultures. PMID- 11501413 TI - Effect of starvation and chloramphenicol on acceleration of bacterial dihexyl sulfosuccinate biotransformation. AB - Starvation for carbon and energy sources accelerated the biotransformation of the anion-active surfactant dihexyl sulfosuccinate (DHS) by Comamonas terrigena cells. Chloramphenicol (Cm) added at different time intervals to non-starved cells inhibited the DHS transformation. The largest difference between cells treated and non-treated by Cm was observed for a 16-h-starvation period. Protein synthesis de novo during starvation enhanced the DHS biotransformation efficiency. A partial transformation of DHS in the presence of Cm indicated the constitutive character of enzymes involved in primary DHS biodegradation. PMID- 11501414 TI - Stable chloroplast transformation in Chlamydomonas reinhardtii using microprojectile bombardment. AB - The chloroplasts of Chlamydomonas reinhardtii were transformed using a vector (paadAGUS4.1) that contained a spectinomycin-resistance gene (aadA) as a selectable gene, and bacterial uidA (GUS) as a reporter gene, and pea 4.1 kb D loop containing sequence. The vector was introduced into the alga through particle gun bombardment. The transformed colonies were screened for the presence of foreign genes by Southern hybridization using GUS, aadA and 4.1 pea Ori probes. Expression of aadA and GUS genes was detected in all colonies that were grown on spectinomycin. A detailed restriction analysis followed by southern hybridization of total genomic DNA using pea 4.1 kb D-loop as probe indicated that the D-loop sequence can serve in site-specific integration of foreign DNA due to high homology. Restriction analysis of different colonies showed that the foreign DNA was probably present in a mixture population of autonomous segment and integrated in the native chloroplast genome. PMID- 11501415 TI - Effects of a ferrate-containing preparation on diverse metabolic processes in yeast. AB - A plant-sap-derived preparation containing bi- and tervalent ferrate anions was tested on growth, respiration on glucose, and membrane transport of 6-deoxy-D glucose (6-dGlc) and 2-aminoisobutyric acid (Aib) in several yeast species, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Lodderomyces elongisporus, Rhodotorula gracilis, and Dipodascus magnusii. Growth was enhanced by as much as 65%, respiration was not affected significantly except for a decrease in R. gracilis, transport of 6-dGlc was not affected while that of Aib was increased by up to 45% in R. gracilis and up to 27% in L. elongisporus. PMID- 11501416 TI - Spontaneous and radical-induced plasma membrane lipid peroxidation in differently oxidant-sensitive yeast species and its suppression by antioxidants. AB - Formation of thiobarbituric acid-reactive substances (TBRS; nmol/mg lipids) indicative of lipid peroxidation was measured in whole cells and in isolated plasma membrane lipids from three yeast species differing in oxidant sensitivity (Schizosaccharomyces pombe, Saccharomyces cerevisiae and Rhodotorula glutinis) after exposure to the Fenton reagent, FeII, H2O2, tert-butyl hydroperoxide (TBHP) and azo compounds (AAPH, ACHN). In whole cells, spontaneous TBRS formation rose in the sequence S. pombe < S. cerevisiae < R. glutinis (1:approximately 5:approximately 7). Oxidants increased the TBRS production 13-18 fold in the sequence FeII approximately TBHP > AAPH approximately ACHN approximately Fe Fenton > H2O2. This increase need not be solely due to increased lipid peroxidation. In isolated plasma membrane lipids from all three species, the spontaneous TBRS production referred to 1 mg lipids was 9-13-fold higher than in whole cells. In S. pombe lipids, only TBHP increased the TBRS production. In lipids from S. cerevisiae and R. glutinis, all added oxidants increased the spontaneous TBRS production 2-3 times in the sequence TBHP > ACHN > AAPH > FeII > Fe-Fenton > H2O2. Oxidant-induced TBRS production in both whole cells and isolated membrane lipids was partially suppressed by the lipid peroxidation inhibitors 2,6-di-tert-butyl-4-methylphenol ("butylated hydroxytoluene"; BHT) and the newly synthesized PYA12 compound. Both agents were more effective in isolated lipids than in whole cells and against OH.-producing than against ROO.- or RO. producing oxidants. Yeast membrane lipids, which are generally poor in polyunsaturated fatty acids, are thus subject to perceptible lipid peroxidation. PMID- 11501417 TI - Effects of the Fenton reagent on transport in yeast. AB - In the facultatively anaerobic yeast Saccharomyces cerevisiae the uptake rate and the accumulation ratio of 2-aminoisobutyric acid was decreased by some 30% by Fenton's reagent (FR), a powerful source of OH. radicals. Likewise, the uptake of glutamic acid, leucine and arginine was diminished. The mediated diffusion of 6 deoxy-D-glucose was not affected. The H+ symport of maltose and trehalose was inhibited by some 40% both in the initial rate and in the accumulation ratio. FR had a dramatic inhibitory effect when present during preincubation with 50 mmol/L glucose. In the obligately aerobic Lodderomyces elongisporus the uptake of all amino acids tested was decreased by 15-30%, that of 6-deoxy-D-glucose by about 10%. The initial rates of uptake of maltose and trehalose were depressed by FR by 40% and the acceleration of uptake observed after 8 min of incubation, was abolished by FR completely. Acidification rate of the external medium by S. cerevisiae in the presence of glucose or galactose was enhanced three-fold, that after subsequently added K+ was substantially decreased. FR appears to have a dual effect on sugar and amino acid transport processes in yeast: (1) it blocks carrier protein synthesis; (2) it inhibits the source of energy for transport. It does not appreciably affect the carrier proteins themselves. PMID- 11501418 TI - Sugar repression in the methylotrophic yeast Hansenula polymorpha studied by using hexokinase-negative, glucokinase-negative and double kinase-negative mutants. AB - Two glucose-phosphorylating enzymes, a hexokinase phosphorylating both glucose and fructose, and a glucose-specific glucokinase were electrophoretically separated in the methylotrophic yeast Hansenula polymorpha. Hexokinase-negative, glucokinase-negative and double kinase-negative mutants were isolated in H. polymorpha by using mutagenesis, selection and genetic crosses. Regulation of synthesis of the sugar-repressed alcohol oxidase, catalase and maltase was studied in different hexose kinase mutants. In the wild type and in mutants possessing either hexokinase or glucokinase, glucose repressed the synthesis of maltase, alcohol oxidase and catalase. Glucose repression of alcohol oxidase and catalase was abolished in mutants lacking both glucose-phosphorylating enzymes (i.e. in double kinase-negative mutants). Thus, glucose repression in H. polymorpha cells requires a glucose-phosphorylating enzyme, either hexokinase or glucokinase. The presence of fructose-phosphorylating hexokinase in the cell was specifically needed for fructose repression of alcohol oxidase, catalase and maltase. Hence, glucose or fructose has to be phosphorylated in order to cause repression of the synthesis of these enzymes in H. polymorpha suggesting that sugar repression in this yeast therefore relies on the catalytic activity of hexose kinases. PMID- 11501419 TI - Some tryptophan pathways in the phytopathogen Xanthomonas oryzae pv. oryzae. AB - Xanthomonas oryzae pv. oryzae, the causal organism of bacterial blight of rice which produces leaf blight as well as kresek (wilt) symptoms in plants were tested for indole, auxin production in culture supplemented with L-tryptophan. On the basis of indoleacetic acid (IAA) production the isolates were grouped into IAA-positive and IAA-negative. Out of 17 isolates, 11 were IAA-positive while 6 were IAA-negative. The isolates metabolized tryptophan through two different routes and the isolates vary in the pathway of tryptophan utilization. The IAA positive isolates converted tryptophan to IAA as the end product, whereas the IAA negative isolates formed anthranilate as an intermediate metabolite and finally produced pyrocatechol via the kynurenine pathway. Quantification of tryptophan metabolism revealed that the maximum production of IAA and pyrocatechol in culture occurred during 2-d incubation at 30 +/- 2 degrees C. PMID- 11501420 TI - Plant growth promotion and fungicidal activity in a siderophore-producing strain of Proteus sp. AB - A proteus strain inhibited mycelial growth of Fusarium oxysporum in vitro. Seed bacterization showed significant plant growth promotion and Fusarium-wilt suppression activity of Phaseolus mungo in a gnotobiotic system. The culture filtrate of this strain exhibited three prominent bands in UV-VIS spectra between 300 and 400 nm. The growth promotion assay of the extracted compound against different indicator organisms indicated the production of a compound related to a 2-oxoacid-type siderophore. The HPLC of the purified ethyl acetate extract of the strains and standard 4-methyl-2-oxopentanoate (2-oxoisocaproate) revealed a single peak, similarly as the coinjection of the extract and the standard. The production of siderophore, probably 2-oxoisocaproate, was demonstrated. PMID- 11501421 TI - Effect of soil bacteria on hyphal growth of the arbuscular mycorrhizal fungus Glomus claroideum. AB - The effect of 46 bacterial strains isolated from tilled and non-tilled soils collected at 3 localities on the growth of intraradical hyphae of the arbuscular mycorrhizal (AM) fungus Glomus claroideum was demonstrated. A larger number of stimulatory bacterial isolates was obtained from tilled soils, but the bacteria showing the strongest stimulation of hyphal growth were isolated from a soil that had not been cultivated. Isolates obtained from hyphae of AM fungi showed no substantial stimulatory effects, but produced more uniform effects on hyphal growth than the isolates of bacteria obtained from soil. Bacterial cenoses present in 3 different soils differ significantly in their effects on AM fungi. PMID- 11501422 TI - Antifungal activity of Bacillus sp. isolated from compost. AB - Four strains of Bacillus isolated from lupine compost exhibited an antifungal activity against six plant fungal pathogens (Rhizoctonia solani, Bipolaris sorokiniana, Sclerotinia sclerotiorum, Trichothecium roseum, Fusarium solani, Fusarium oxysporum). It was significantly influenced by the composition of the cultivation media. PMID- 11501423 TI - Antibiotic resistance patterns of coliforms isolated from six protected wetlands in the southeast of Spain. AB - Coliforms and thermotolerant coliforms were isolated from six protected lakes in the Antequera area of Spain in order to assess the sanitary quality of these wetlands. The thermotolerant coliform group consisted of several bacterial genera with a wide spectrum of antibiotic resistance. Resistance among Escherichia coli strains was correlated with the origin of the sample and three homogeneous groups of wetlands were distinguished. One of them, which corresponded to wetlands where faecal indicators were indigenous, exhibited no correlation between these bacteria and faecal pollution. Non-coli strains showed a similar range of antibiotic susceptibility for this group. In the other two groups a faecal pollution was detected and antibiotic resistance for E. coli strains established the origin of pollution. PMID- 11501424 TI - Relative pathogenicity of Candida tropicalis in rat tongue mucosa. AB - The potential of C. tropicalis to colonize and infect rat tongue mucosa was demonstrated. Thirty female Sprague-Dawley rats were infected orally with three different strains of C. tropicalis. The animals were killed one and three weeks following the inoculation and sections of their tongue were stained with hematoxylin-eosin and Grocott stain. Histological changes were observed in the group of animals killed one week after inoculation and infected with C. tropicalis strain isolated from the crural ulcer of a diabetic patient. The most important finding was acute purulent myositis with the formation of abscesses. The myositis was local without signs of spreading to the surrounding tissue. Epithelium-penetrating hyphae observed in the Grocott-stained sections were relatively fewer and more sparsely distributed. PMID- 11501425 TI - Influence of intoxication with organophosphates on rumen bacteria and rumen protozoa and protective effect of clinoptilolite-rich zeolite on bacterial and protozoan concentration in rumen. AB - The effect of O-ethyl-S-(2-diisopropylaminoethyl) methylthiophosphonate on rumen bacteria and rumen protozoa was investigated in sheep (after premedication with clinoptilolite-rich zeolite and without that premedication). In control animals a decrease in the total concentration of rumen protozoa was observed 3-7 d after intoxication (particularly in small and large ones). In clinoptilolite-rich zeolite-treated animals only a slight decrease in protozoan numbers occurred during the first hours after the intoxication. Similarly, in every category of rumen bacteria marked differences between the groups were recorded, particularly in concentration of lipolytic bacteria. The results suggest some protective effect of clinoptilolite-rich zeolite for rumen microbiota against the organophosphate poison. PMID- 11501426 TI - Degradation of polycyclic aromatic hydrocarbons by the copper(II)-hydrogen peroxide system. AB - A non-enzymic system containing CuSO4 (10 mmol/L) and hydrogen peroxide (100 mmol/L) was used for the degradation of three polycyclic aromatic hydrocarbons: phenanthrene, fluoranthene, and pyrene (all at 10 mmol/L). The system degraded the compounds rapidly and efficiently. After 1 d at room temperature, more than 80% of pyrene, phenanthrene, and fluoranthene disappeared. Several products are formed during the reaction including a black precipitate. PMID- 11501427 TI - Characterization of the third generation enzyme immunoassay IEA-HIV1/2-III for the detection of anti-HIV specific antibodies in human sera. AB - The sensitivity and specificity of the developed anti-HIV1/2 third generation enzyme immunoassay, the IEA-HIV1/2-III, was examined. The test system for the detection of anti-HIV antibodies included peroxidase-conjugated HIV-specific recombinant Gag protein fragments (epitopes of p24 and p17 proteins), Env-1 (epitopes of p41 and p120 proteins), and Env-2 (p36 epitopes). Sensitivity was evaluated with 346 sera from HIV1-seropositive subjects, Anti-HIV1 Low Titer panels no. 10 and PRB-106 and seropositive panel PRB-931 in comparison with other third- and second-generation assays. The IEA-HIV1/2-III assays are characterized with high sensitivity comparable to the other third generation assays and the better sensitivity with respect to the second generation test-kit to determine HIV-specific antibodies in human sera. The specificity was determined using three hundred sixty-seven potentially cross-reactive samples (but negative for anti HIV1/2). Only one specimen among them was reactive by IEA-HIV1/2-III. PMID- 11501428 TI - Genetic markers and genotyping methodologies: an overview. PMID- 11501429 TI - Sublethal identification of susceptible individuals: using swim performance to identify susceptible fish while keeping them alive. AB - One branch of ecotoxicology has focused on identifying genetic markers in fish that are associated with susceptibility to toxic compounds. In laboratory studies, a common approach has been to compare the genetic variation in fish that die first in time-to-death studies to that found in fish that live longer or survive the exposure. Studies of this kind would benefit from the ability to identify living individuals as susceptible, as these individuals could then be used to answer currently unanswerable questions. The purpose of this mini-review is to suggest that post-exposure swim performance can be used to sublethally discriminate between susceptible and resistant individual fish after these fish have been exposed to environmental stressors, particularly heavy metals. PMID- 11501430 TI - Consequences of polluted environments on population structure: the bank vole (Clethrionomys glareolus) at Chornobyl. AB - The accident at the Chornobyl Nuclear Power Plant in April 1986, released 100-200 million Curies of radioactive material into the surrounding environment. To investigate the possible genetic and population effects resulting from chronic exposure to this environmental radiation, we have examined mitochondrial DNA (control region) sequences from bank voles, Clethrionomys glareolus, inhabiting contaminated sites. Our analysis indicates genetic diversity is elevated in the contaminated sites when compared to relatively uncontaminated reference sites. This may be attributed to either an increased mutation rate in the mtDNA control region or immigration of individuals from surrounding areas into the contaminated environment. Although our observations do indicate that the contaminated areas represent sink populations, we cannot statistically discriminate between these two alternatives at this time. In addition, we have been unable to attribute any significant detrimental effects to bank vole populations inhabiting the contaminated Chornobyl environment based on these data. This is particularly paradoxical considering bank voles in the contaminated areas harbor the highest radiocesium (137Cs) body burdens and external dose rates of any mammal ever measured. Our long-term research on the bank vole indicates that several factors, including contaminants, may affect haplotype dynamics both spatially and temporally. These multifarious influences subsequently affect population genetic estimates typically used to address the effects of environmental pollution on animal populations. Finally, we provide a general framework for designing experiments investigating the role contaminants play in altering the genetic characteristics of exposed populations. PMID- 11501431 TI - Ecotoxicology and population genetics: the emergence of "phylogeographic and evolutionary ecotoxicology". AB - Genetics of ecotoxicology has recently emerged as a priority research field. The advent of polymerase chain reaction and molecular population genetics has made it possible to examine the genetics in even the smallest individuals. Although a potentially powerful technique, current approaches oversimplify the relationship of change in gene frequency to contaminant exposure. Many of these approaches cannot control for random correlation or accessory abiotic factors that impinge on the system tested. Indeed, the gestalt approaches of laboratory exposure or natural field experiments may ignore significant genome-level interactions that are important within a given system. At the very least, these approaches would benefit by a biogeographic survey of genetic variation to understand geographic microevolutionary patterns, or phylogeography, within a species to reduce spurious correlations and erroneous conclusions. Other single locus approaches can be chosen to enhance this approach if genetic/environmental interactions have been characterized for laboratory populations or for other model systems. PMID- 11501432 TI - Population genetic structure of the western mosquitofish, Gambusia affinis, in a highly channelized portion of the San Antonio River in San Antonio, TX. AB - Population genetic assessments were performed on populations of the mosquitofish, Gambusia affinis, sampled at seven sites along a 10 km reach of the upper San Antonio River in San Antonio, TX. Mosquitofish populations were sampled from the downtown area, known as the "Riverwalk", where the river is concrete lined, deeply pooled, and receives heavy tour boat traffic. Populations were also sampled from sites upstream and downstream from the Riverwalk. Allozyme electrophoresis was performed on approximately 50 individuals from each site. Five of 12 loci surveyed were polymorphic (95% criterion). Exact tests over all loci for all populations indicated that allele frequencies differed significantly among sites. Allele frequencies of the upstream population were significantly different from the Riverwalk populations. Differences resulted primarily from significant decreases in frequencies of rare alleles at MDH-2* and two GPI* loci in the downtown area. Mean dissolved oxygen measurements were also significantly lower at Riverwalk sites. These results suggest that selective forces such as non point source runoff or low dissolved oxygen, perhaps in combination with limited migration due to numerous dams, have reduced genetic diversity of populations in the downtown area. PMID- 11501433 TI - Development of DNA-based microsatellite marker technology for studies of genetic diversity in stressor impacted populations. AB - In recent years microsatellite marker applications in various disciplines increased rapidly. Microsatellites are characterized as independent and highly variable markers most noted for their usefulness and popularity among researchers. Microsatellites can be constructed in-house from a partial genomic library, ordered from a vendor, or "borrowed" from a species that is closely related to the target species. Microsatellite application in natural populations can reveal present as well as historical impacts of stressors to particular population. PMID- 11501434 TI - Development and use of microsatellite DNA loci for genetic ecotoxicological studies of the fathead minnow (Pimephales promelas). AB - The development of new DNA technologies has increased the opportunities to study the effect of genetic variation as a factor in selection, resistance, and fitness of individuals and populations. Genetic composition affects the evolutionary potential of an organism; however, whether genetic variation has a role in adaptation and survival due to chemical stress is still under debate. The strategy of measuring such variation should be based on the desired number of loci needed and degree of polymorphism at each locus. Microsatellite DNA loci (also known as Simple Tandem Repeats, STRs) have among the highest levels of variation within most eukaryotic genomes. Methods to determine the high levels of variation in individuals and populations have become more refined, making DNA based studies in ecotoxicology more feasible. Herein, we describe the general characteristics of microsatellite DNA loci, their isolation, and their applications to toxicological studies using the fathead minnow (Pimephales promelas) as a focal species. PMID- 11501435 TI - Choice of methodology for assessing genetic impacts of environmental stressors: polymorphism and reproducibility of RAPD and AFLP fingerprints. AB - PCR-based multi-locus DNA fingerprints represent one of the most informative and cost-effective measures of genetic diversity and are useful population-level biomarkers of toxicologic and other anthropogenic impacts. However, concerns about reproducibility of DNA fingerprints have limited their wider use in environmental biology. We assessed polymorphism and reproducibility of two common fingerprinting techniques, RAPD (randomly amplified polymorphic DNA) and AFLP (amplified fragment length polymorphism), in pedigreed populations of rainbow trout (Oncorhynchus mykiss) to derive general rules for selective removal of problematic fingerprint bands. We found that by excluding bands that comprised less than 1% of total intensity, and by excluding the largest and smallest 10% of the bands, we could achieve nearly 100% reproducibility of AFLP fingerprints. Similar application of band exclusion criteria to RAPD fingerprints did not significantly enhance their reproducibility, and at least 15% of RAPD bands were not fully repeatable, heritable, or transmittable. The RAPD technique produced more polymorphic fingerprints than AFLP; however, considering that a substantial proportion of RAPD markers did not demonstrate Mendelian inheritance patterns, the AFLP methodology is to be preferred for future research. PMID- 11501436 TI - Integration of genotoxic and population genetic endpoints in biomonitoring and risk assessment. AB - Genetic ecotoxicology is a multifaceted discipline that examines the effects of xenobiotic compounds on the structure and function of DNA. This paper discusses the role of genetic ecotoxicology in environmental biomonitoring and risk assessment. Genetic ecotoxicology may include somatic effects (e.g., DNA damage) or population genetic effects (changes in genetic diversity or gene frequencies). Traditionally, genetic ecotoxicology studies have focused on either one of these sub-disciplines, but integration of these two approaches would be advantageous for three reasons. First, at the population level, concordant responses between changes in population genetic structure and elevated levels of DNA damage may provide evidence that the population genetic changes are influenced by exposure to genotoxic chemicals. Second, if the frequencies of alleles or other genetic markers differ between genotoxicant-contaminated and reference populations, associations between relative amount of DNA damage and genotype may provide evidence that these changes are due to genotoxicant-induced selection. Third, genetic analysis of gene flow may provide insight into patterns of dispersal that could obscure differences between contaminated and reference populations. In order to demonstrate the application of these ideas, three lines of research are summarized herein. The first is a series of studies that focus on radionuclide contaminated populations of mosquitofish (Gambusia). This research identified RAPD markers that may be indicative of genetic adaptation to radionuclide stress. Relative amounts of DNA damage among genotypes presented evidence that these markers may be indicators of relative radioresistance. The second study examined DNA damage and population genetic structure in radionuclide-contaminated kangaroo rat (Dipodomys) populations. It was found that between-population differences in genetic diversity paralleled those for DNA damage and relative levels of contamination. Also, population genetic analysis indicated that there was dispersal between contaminated and reference populations, and that between population differences in the amount of DNA damage could not be detected until this dispersal was taken into account. In the third study, populations of redbreast sunfish (Lepomis auritris) from streams contaminated with complex mixtures of industrial chemicals were examined. It was found that the genetic distances between populations within the contaminated stream corresponded with the relative magnitude of molecular and community-level effects. It was concluded that genetic ecotoxicology could make significant contributions to the fields of environmental biomonitoring and ecological risk assessment, and that integration of genotoxicology and population genetic studies would be a definite advantage toward this end. PMID- 11501437 TI - What makes recovered-memory testimony compelling to jurors? AB - Little is known about how jurors arrive at verdicts in cases involving recovered memories of childhood sexual abuse. Study 1 investigated mock jurors' reactions to the recovered-memory testimony of an alleged victim when a therapist intervened with hypnosis, suggestion, or symptom management. When a therapist used hypnosis, jurors viewed the victim's recovered-memory testimony as particularly accurate and credible, and favored the victim in their verdicts. In Study 2, mock jurors were presented with a therapist who was sued for allegedly influencing a client's recall of false memories of abuse. In this case, however, jurors viewed therapists who used hypnosis or suggestion as more likely to have created false memories, more responsible for having caused harm, and less competent, and tended not to favor these therapists in their verdicts. We discuss these seemingly contradictory findings in terms of how culturally formed expectancies about hypnosis produce different causal explanations depending on the focus of a trial. PMID- 11501438 TI - Detecting deception in children's testimony: factfinders' abilities to reach the truth in open court and closed-circuit trials. AB - This study examined the influence of closed-circuit television (CCTV) on jurors' abilities to detect deception in children's testimony. Children ages 7-9 individually played games and made a video movie with a male confederate. In the guilty condition, stickers were placed on exposed body parts (i.e., the child's arm, toes, and bellybutton). In the not-guilty and deception conditions, stickers were placed on the child's clothing rather than on bare skin. Approximately 3 weeks later, mock jurors recruited from the community viewed child participants testify either in a traditional courtroom setting or via one-way CCTV. The mock jurors responded to questions about the child witness and the defendant as well as deliberated to reach a verdict. Children in the deception condition were asked to testify as if the stickers had been placed on exposed body parts rather than on their clothing. Predeliberation, jurors were less likely to convict when a child testified in the deception condition as opposed to the guilty condition. These differences disappeared following deliberation. There was no support for the notion that jurors reach the truth better when children testify in open court versus via CCTV. Implications for jurors' abilities to reach the truth are discussed. PMID- 11501439 TI - The effects of note-taking and trial transcript access on mock jury decisions in a complex civil trial. AB - Mock juries were either permitted to take notes or not and provided with access to the trial transcript during deliberations or were not given access. Juries viewed a videotape of a complex trial involving multiple plaintiffs. Note-taking juries were able to distinguish among differentially worthy plaintiffs when assigning awards while non note takers did not distinguish among the plaintiffs and allocated higher overall compensation. Note-taking was significantly more effective than access to trial transcripts in increasing jury competence. Note taking juries appeared better able to recognize probative evidence and reject false lures than were non note-taking juries. Limits and implications of the present study were discussed. PMID- 11501440 TI - The influence of actuarial risk assessment in clinical judgments and tribunal decisions about mentally disordered offenders in maximum security. AB - Research has shown that actuarial assessments of violence risk are consistently more accurate than unaided judgments by clinicians, and it has been suggested that the availability of actuarial instruments will improve forensic decision making. This study examined clinical judgments and autonomous review tribunal decisions to detain forensic patients in maximum security. Variables included the availability of an actuarial risk report at the time of decision making, patient characteristics and history, and clinical presentation over the previous year. Detained and transferred patients did not differ in their actuarial risk of violent recidivism. The best predictor of tribunal decision was the senior clinician's testimony. There was also no significant association between the actuarial risk score and clinicians' opinions. Whether the actuarial report was available at the time of decision making did not alter the statistical model of either clinical judgments or tribunal decisions. Implications for the use of actuarial risk assessment in forensic decision making are discussed. PMID- 11501441 TI - Evaluating the comprehensibility of jury instructions: a method and an example. AB - Methodological problems in jury simulation research involve issues of sampling, choice of stimulus materials, appropriate unit of analysis, appropriate dependent variable, corroborative data, and problems of role playing. Despite these issues, comprehension of jury instructions may be suitable for examination by jury simulation techniques--if certain of these methodological concerns can be satisfied. In a series of 5 experiments using typical Canadian legal instructions on criminal conspiracy and the coconspirator exception to the hearsay rule, this study attempted to validate a simple and inexpensive technique for testing the incomprehensibility of a given set of jury instructions by requiring participants to apply those legal instructions to a set of facts. The results demonstrate the utility of an application test, and suggest that for assessing the comprehensibility of jury instructions, it may be acceptable to use undergraduate students as participants, to use individual participants without group deliberation, and to employ written stimulus materials. PMID- 11501443 TI - Physician, know thy limits. PMID- 11501442 TI - Time to weed the CPG garden. PMID- 11501444 TI - Physicians, know thy limits. PMID- 11501445 TI - Do women treated for breast cancer at teaching hospitals really fare better? PMID- 11501446 TI - Do women treated for breast cancer at teaching hospitals really fare better? PMID- 11501447 TI - Do women treated for breast cancer at teaching hospitals really fare better? PMID- 11501448 TI - Do women treated for breast cancer at teaching hospitals really fare better? PMID- 11501449 TI - Do women treated for breast cancer at teaching hospitals really fare better? PMID- 11501450 TI - Do women treated for breast cancer at teaching hospitals really fare better? PMID- 11501451 TI - Raloxifene: handle with care. PMID- 11501452 TI - Nonclinical factors in patient selection for surgery. PMID- 11501453 TI - Lifestyle drugs. PMID- 11501454 TI - What is the quality of drug therapy clinical practice guidelines in Canada? AB - BACKGROUND: The Canadian Medical Association maintains a national online database of clinical practice guidelines developed, endorsed or reviewed by Canadian organizations within 5 years of the current date. This study was designed to identify and describe guidelines in the database that make recommendations related to the use of drug therapy, and to assess their quality using a standardized guideline appraisal instrument. METHODS: Drug therapy guidelines in the database were identified with the use of search terms and hand searching. Descriptive information about the developers, endorsement by other organizations, publication status, disease and drug focus was abstracted. Each guideline was independently assessed by 3 appraisers (a physician, a pharmacist and a methodologist) with the use of the Appraisal Instrument for Clinical Guidelines. Conditions were classified according to the tenth revision of the International Statistical Classification of Diseases and Related Health Problems. RESULTS: We identified 217 drug therapy guidelines produced or reviewed from 1994 to 1998. Guideline developers included national organizations (47.0%), paragovernment organizations (39.6%) and professional associations (30.9%); 31.3% of the guidelines were published, and 10.6% stated drug company sponsorship. The most common conditions addressed by the guidelines were infections and parasitic diseases (39.6%), neoplasms (11.5%) and diseases of the circulatory system (11.5%). Drugs most commonly cited were anti-infective agents (42.9%), antiviral agents (15.2%) and cardiovascular drugs (16.1%). Eleven organizations produced 176 (81.1%) of the guidelines. In all, 14.7% of the guidelines met half or more of the 20 items assessing rigour of guideline development on the appraisal instrument (mean quality score 30.0% [95% confidence interval (CI) 27.5%-32.6%]), 61.8% met half or more of the 12 items assessing guideline context and content (mean score 57.0% [95% CI 54.6%-59.3%]), and none met half or more of the 5 items assessing guideline application (mean score 5.6% [95% CI 4.7%-6.5%]). Overall, 64.6% of the guidelines were recommended with modification by at least 2 of the 3 appraisers, 9.2% were recommended without change, and 26.3% were not recommended. The quality of the guidelines assessed varied significantly by developer, publication status and drug company sponsorship. No substantial improvement in guideline quality was observed over the 5-year study period. INTERPRETATION: Developers of Canadian drug therapy guidelines are producing guidelines that are often perceived to be clinically useful to physicians and pharmacists, although the methods (or the description of the methods) by which they are developed need to be more rigorous and thorough. PMID- 11501455 TI - Clinical practice guidelines for the care and treatment of breast cancer: 13. Sentinel lymph node biopsy. AB - OBJECTIVE: To provide information and recommendations to women with breast cancer and their physicians regarding what is now known about sentinel lymph node (SLN) biopsy. OPTIONS: Axillary dissection; SLN biopsy followed by backup axillary dissection; SLN biopsy. OUTCOMES: Accurate determination of cancer stage, resulting in better-informed therapeutic decisions. EVIDENCE: Systematic review of English-language literature published from January 1991 to December 2000 retrieved primarily from MEDLINE and CANCERLIT. RECOMMENDATIONS: Axillary dissection is the standard of care for the surgical staging of operable breast cancer. If a patient requests or is offered SLN biopsy, the benefits and risks as well as what is and is not known about the procedure should be outlined. Patients should be informed of the number of SLN biopsies performed by the surgeon and the surgeon's success rate with the procedure, as determined by the identification of the SLN and the false-negative rate (the presence of tumour cells in the axillary nodes when the SLN biopsy result is negative). Before surgeons replace axillary dissection by SLN biopsy as the staging procedure at their institution, they should (a) familiarize themselves with the literature on the topic and the techniques needed to perform the procedure, (b) follow a defined protocol for all 3 aspects of the procedure (nuclear medicine, surgery, pathology) and (c) perform backup axillary dissection until an acceptable success rate (as determined by the identification of the SLN and the false-negative rate) is achieved. A surgeon who performs breast cancer surgery infrequently should not perform SLN biopsy. A positive SLN biopsy result or failure to identify an SLN should prompt full axillary dissection. SLN biopsy is contraindicated in women who have clinically palpable nodes, locally advanced breast cancer, multifocal tumours, previous breast surgery or previous irradiation of the breast. Staining of tissue sections with hematoxylin and eosin, and not immunohistochemical analysis for cytokeratin, should determine adjuvant therapy. Participation in randomized clinical trials is encouraged. [A patient version of these guidelines appears in Appendix 1.] VALIDATION: Internal validation within the Steering Committee on Clinical Practice Guidelines for the Care and Treatment of Breast Cancer; no external validation. PMID- 11501456 TI - Does how you do depend on how you think you'll do? A systematic review of the evidence for a relation between patients' recovery expectations and health outcomes. AB - BACKGROUND: Most clinicians would probably agree that what patients think will happen can influence what does happen over the clinical course. Yet despite useful narrative reviews on expectancy of therapeutic gain and the mechanisms by which expectancy can affect health outcomes, we were unable to locate a systematic review of the predictive relation between patients' recovery expectations and their health outcomes. METHODS: We searched MEDLINE for English language articles published from 1966 to June 1998 with a title or abstract containing at least 1 of the medical subject headings (MeSH) "self-assessment," "self-concept" or "attitude to health," or the MeSH subheading "psychology," and at least 1 word from each of 3 sets: "patient" and similar words; a form of "expectation," "belief" or "prediction"; and a form of "recover," "outcome," "survival" or "improve." Relevant articles contained original research data, measured patients' recovery expectations, independently measured a subsequent health outcome and analyzed the relation between expectations and outcomes. We assessed internal validity using quality criteria for prognostic studies based on 6 categories (case definition; patient selection; extent of follow-up; objective outcome criteria; measurement and reporting of recovery expectations; and analysis). RESULTS: A total of 1243 titles or abstracts were identified through the computer search, and 93 full-text articles were retrieved. Forty-one of these articles met the relevance criteria, along with 4 additional articles identified through other means. Agreement beyond chance on quality assessments of 18 randomly selected articles was high (kappa = 0.87, p = 0.001). Sixteen of the 45 articles provided moderate-quality evidence and included a range of clinical conditions and study designs; 15 of the 16 showed that positive expectations were associated with better health outcomes. The strength of the relation depended on the clinical conditions and the measured used. INTERPRETATION: Consistency across the studies reviewed and the evidence they provided support the need for clinicians to clarify patients' expectations and to assist them in having appropriate expectations of recovery. The understanding of the nature, extent and clinical implications of the relation between expectations and outcomes could be enhanced by more conceptually driven and methodologically sound research, including evaluations of intervention effectiveness. PMID- 11501457 TI - Further disquiet on the guidelines front. PMID- 11501458 TI - Promoting effective guideline use in Ontario. PMID- 11501459 TI - Rheumatology: 16. Diagnosing musculoskeletal pain in children. PMID- 11501460 TI - Bioethics for clinicians: 27. Catholic bioethics. AB - There is a long tradition of bioethical reasoning within the Roman Catholic faith, a tradition expressed in scripture, the writings of the Doctors of the Church, papal encyclical documents and reflections by contemporary Catholic theologians. Catholic bioethics is concerned with a broad range of issues, including social justice and the right to health care, the duty to preserve life and the limits of that duty, the ethics of human reproduction and end-of-life decisions. Fundamental to Catholic bioethics is a belief in the sanctity of life and a metaphysical conception of the person as a composite of body and soul. Although there is considerable consensus among Catholic thinkers, differences in philosophical approach have given rise to some diversity of opinion with respect to specific issues. Given the influential history of Catholic reflection on ethical matters, the number of people in Canada who profess to be Catholic, and the continuing presence of Catholic health care institutions, it is helpful for clinicians to be familiar with the central tenets of this tradition while respecting the differing perspectives of patients who identify themselves as Catholic. PMID- 11501461 TI - Don't panic, it's organic: dispute over natural health products regulations continues. PMID- 11501462 TI - No public voice in genetics research? PMID- 11501463 TI - BC's liberals promise a revolution, with health care one of the targets. PMID- 11501464 TI - Putting a human face on neurologic illness. PMID- 11501465 TI - Methylmercury exposure: fishing for answers. PMID- 11501466 TI - Colorectal cancer screening. Recommendation statement from the Canadian Task Force on Preventive Health Care. PMID- 11501467 TI - Thermostable glucose-tolerant glucoamylase produced by the thermophilic fungus Scytalidium thermophilum. AB - Glucoamylase produced by Scytalidium thermophilum was purified 80-fold by DEAE cellulose, ultrafiltration and CM-cellulose chromatography. The enzyme is a glycoprotein containing 9.8% saccharide, pI of 8.3 and molar mass of 75 kDa (SDS PAGE) or 60 kDa (Sepharose 6B). Optima of pH and temperature with starch or maltose as substrates were 5.5/70 degrees C and 5.5/65 degrees C, respectively. The enzyme was stable for 1 h at 55 degrees C and for about 8 d at 4 degrees C, either at pH 7.0 or pH 5.5. Starch, amylopectin, glycogen, amylose and maltose were the substrates preferentially hydrolyzed. The activity was activated by 1 mmol/L Mg2+ (27%), Zn2+ (21%), Ba2+ (8%) and Mn2+ (5%). Km and vlim values for starch and maltose were 0.21 g/L, 62 U/mg protein and 3.9 g/L, 9.0 U/mg protein, respectively. Glucoamylase activity was only slightly inhibited by glucose up to a 1 mol/L concentration. PMID- 11501468 TI - Steroid biotransformation by different strains of Micrococcus sp. AB - A strain of Micrococcus sp. was isolated for its capability of side chain degradation of cholesterol. This strain was characterized and identified as Micrococcus roseus. It was found to be the best strain for the production of androsta-1,4-diene-3,17-dione and androst-4-ene-3,17-dione compared with other Micrococcus strains. PMID- 11501469 TI - OR-1--a mixture of esters of glyceric acid produced by Penicillium funiculosum and its antitrypsin activity. AB - A mixture of related metabolites (denoted OR-1) was isolated from the fermentation broth of Penicillium funiculosum together with mitorubrinic acid. Structurally OR-1 is glyceric acid esterified with C14-C18 fatty acids. Steady state studies revealed that OR-1 behaved like an uncompetitive trypsin inhibitor with Ki 17.6 mumol/L. PMID- 11501470 TI - Survival of and wheat-root colonization by alginate encapsulated Herbaspirillum spp. AB - The survival of Herbaspirillum spp. cells added directly or encapsulated in alginate beads and colonization of wheat roots was evaluated in soil microcosms. Cells entrapped in alginate in the presence of JNFb-broth and introduced into unplanted non-sterile clay loamy and sandy soils survived better than cells added directly to the same soils after 50 d incubation. On amendment by JNFb broth and/or skim milk the entrapped cells survived better than those prepared in water. Encapsulated cells survived better in a heavier textured soil (clay-loamy) than in a lighter (sandy) soil. Wheat plants growing in microcosms inoculated with various bead types from day 0 to day 30 exhibited high levels of histosphere colonization, nitrogenase activity (in situ) measured by acetylene reduction assay, plant dry mass and total N content but no symptoms of mottled stripe disease were observed. Comparable results of growth criteria and nitrogenase activity, but relatively lower bacterial populations, were obtained with wheat grown for 45 d after the inoculant had been introduced into the soil with different bead types. PMID- 11501471 TI - Kinetic properties of fractions of extracellular NAD+ nucleosidase from Streptococcus pyogenes as an example of host selection by a pathogen: possible role of serum albumin in the organism. AB - Preparative isoelectric focusing was used to separate free bacterial NAD+ nucleosidase from its complex with a bound host component. Both fractions were characterized by optimum temperature and activation energy of denaturation. The bacterial product is enzymically inactive. The enzymically active structure is formed upon binding to the host component. Only the host organism can provide the suitable, activating structure. The host component in the present system is added to the cultivation medium with a beef heart extract but it can be replaced by serum albumin. The possible role of albumin as a carrier structure for flexible and enzymically inactive peptides is discussed. Different peptides bound to albumin can provide different enzyme activities. The term binary enzyme is coined, referring to a situation where the two enzyme components are coded at genetically distant loci. The pathogen makes use of the carrier structure of albumin type and produces another polypeptide invested with an enzyme activity convenient for the pathogen. PMID- 11501472 TI - Non-specific DNAases from the rumen bacterium Prevotella bryantii. AB - Extracellular non-specific nucleases were observed in some strains belonging to the ruminal species of the genus Prevotella, mostly P. brevis and P. bryantii. The nuclease from P. bryantii appeared to be extracellular; it mediates the degradation of the supercoiled plasmid DNA via an open circle intermediate. The cleavage is not site specific although a preference for certain cleavage sites does seem to exist. Our attempts to clone the wild-type P. bryantii B(1)4 nuclease in E. coli strain ER1992 that reports on the DNA damage sustained, were unsuccessful probably due to excessive intracellular nuclease activity that killed the cells bearing the gene for the nuclease. On the other hand, the nuclease from a related strain TCl-1, which has a less active enzyme of the same type, was successfully cloned. PMID- 11501473 TI - The bacteriophages of ruminal prevotellas. AB - Rumen bacteriophage-lyzed bacterial strains of the genus Prevotella were isolated and preliminarily characterized. The strain TCl-1 the species P. bryantii was the only prevotella strain successfully infected with filter sterilized rumen fluid from a black-and-white Holstein cow. Two types of plaques were observed, both rather small and turbid. Preliminary electron microscopy observation showed that several morphologically different bacteriophages were present in these plaques. The plaque eluates were further used for the infection of other prevotella strains. The plaques produced by the bacteriophages were observed with two strains, i.e. P. bryantii B(1)4 and P. brevis GA33. The bacteriophages from both strains were examined by transmission electron microscopy and several morphologically different bacteriophages were observed, among others also a large virion with an icosahedral head with the diameter of approximately 120 nm. The bacteriophage was identified in plaques of bacterial cells of the strain GA33 and has an approximately 800 nm long helical tail, which places it among the largest ruminal bacteriophages described to date. Other bacteriophages from the same indicator strain as well as from P. bryantii B(1)4 strain were smaller and tail structures were not observed in all of them. PMID- 11501474 TI - Systematics and evolution of ruminal species of the genus Prevotella. AB - Bacterial species of the genus Prevotella represent a numerically dominant microbial population in the rumen of cattle. They belong to the phylogenetic division Cytophaga-Flexibacter-Bacteroides (CFB) which is a large group of ecologically diverse bacteria with only a few shared traits. The phylogenetic descent from a common ancestor seems to be unquestionable, however, as judged from the small subunit ribosomal RNA analysis. Only 4 ruminal Prevotella species have been described to date, even though the sequence analysis of directly retrieved 16S rRNA genes indicates a large genetic diversity within this group of rumen bacteria. The closest relatives of ruminal Prevotella spp. are not surprisingly other species of the genus Prevotella, typically inhabiting the gastrointestinal tract, oral cavity and genital areas of other animals and man. The previous phylogenetic analysis showed that species of the genus Prevotella can be split into two groups or superclusters, the "ruminal" and the "non-ruminal prevotellas". One of 4 currently described ruminal Prevotella spp., i.e. P. albensis, has been placed outside the supercluster containing ruminal Prevotella spp. and within the supercluster containing the non-ruminal Prevotella spp. However, the number of available small subunit rRNA sequences from this species represents only a fraction of all known ruminal Prevotella sequences. PMID- 11501475 TI - Phenotypic and genetic data supporting reclassification of Butyrivibrio fibrisolvens isolates. AB - Among 55 Butyrivibrio fibrisolvens strains five ribotypes of B. fibrisolvens were described on the basis of RFLP profiles of 16S rDNA regions obtained with restriction endonuclease HaeIII. In the phylogenetic tree, these ribotypes were located in the XIVa cluster of Gram-positive bacteria. Phenotypic differences of selected ribotype groups became the basis for further reclassification of B. fibrisolvens. PMID- 11501476 TI - Production of bacteriocins by different enterococcal isolates. AB - Bacteriocins produced by ruminal as well as environmental isolates were found to be heat-stable antimicrobial substances with a broad inhibitory spectrum (including Gram-negative species and sanitary-important species). The exponential phase of growth and pH 4-7 were optimal for their production. After purification, some of them were sequenced and specified. PMID- 11501477 TI - Application of flow cytometry for ecological monitoring of the rumen microbial ecosystem. AB - Flow cytometry in combination with fluorescently labeled ribosomal RNA oligonucleotide probes was used for enumeration and monitoring of ruminal bacteria. The polyanionic azo dye Trypan Blue was used for discrimination between live bacterial cells and inorganic particles and the separation was further improved by lysozyme treatment and sonication. Cy3-labeled universally conserved probe EUB338 and FITC-labeled Prevotella bryantii specific probe PBB14 were used for in situ hybridization in mixed culture experiments and in samples of crude rumen fluid. The results were analyzed by flow cytometry. The separation of P. bryantii and Butyrivibrio fibrisolvens, another ruminal bacterium, in mixed culture experiments was satisfactory and enabled monitoring of these bacteria in a test system. P. bryantii cells were detected in crude rumen fluid samples only after supplementation with pure culture cells; this implicates a low concentration of P. bryantii cells in vivo (less than 100/nL, i.e. 10(5) per mL). PMID- 11501478 TI - Reliable identification of Prevotella and Butyrivibrio spp. from rumen by fatty acid methyl ester profiles. AB - Data for bacterial identification were provided by culturing anaerobic bacteria under standardized conditions followed by extraction and methylation of cellular long-chain fatty acids and gas chromatographic analysis. The databases of fatty acid methyl ester (FAMEs) profiles for two predominant ruminal genera, Prevotella and Butyrivibrio, were created. Major long-chain cellular fatty acids found in the 23 analyzed Prevotella strains were 15:0 (anteiso), 15:0, 15:0 (iso) and 16:0. The strains of Prevotella could be well identified on species level by the characteristic ratios among major fatty acids and by acids unique fatty for each species. The 45 Butyrivibrio strains were grouped into 4 major and 2 minor groups according to FAMEs profiles. The major fatty acids for the bulk of the Butyrivibrio strains were 14:0, 15:1, 16:0 and 16:0 (iso). This groups corresponded to those based on 16S rDNA sequences. PMID- 11501479 TI - Bacteriocins of ruminal bacteria. AB - Similar sequences of distribution of structural genes encoding enterocin A (isolated from the ruminal strain E. faecium BC25) and enterolysin A (isolated from the ruminal amylolytic strain S. bovis II/1) were demonstrated by PCR using oligonucleotide primers specific for these bacteriocins within the ruminal enterococcal and streptococcal strains. Variable occurrence of these bacteriocins was found within the populations of Gram-positive ruminal cocci. PMID- 11501480 TI - Development of competitive PCR for detection of Butyrivibrio fibrisolvens in the rumen. AB - Competitive PCR method was developed for the detection and enumeration of Butyrivibrio fibrisolvens. Sequences of 16S rDNA were obtained from our isolates (serving as a source of data for primer design) and were distinguished into nine different groups of butyrivibria. Specific primers for two distinct groups were designed with the help of BioEdit program. These primers were tested with DNA of 20 strains of ruminal B. fibrisolvens isolates. Annealing temperature 58 degrees C showed a little specificity but a better selectivity was found after raising it up to 65 degrees C. A group 1 competitive fragment of 16S rDNA of different length was constructed using restriction cutting with MspI followed by ligation; the size of the resulting fragment was cut down by 75 bp. The fragment worked in the presence of the original 16S rDNA fragment of B. fibrisolvens JK 609. PMID- 11501481 TI - Ribosomal RNA genes from Prevotella bryantii: organization and heterogeneity. AB - Five P. bryantii B(1)4 16S rRNA gene copies and their flanking regions were cloned and analyzed. A genomic library was constructed and screened with oligonucleotide DNA probe specific for 16S rRNA gene of P. bryantii. Five out of six different copies of 16S RNA gene were recovered and sequenced. Only minor differences (0.3-1.2%) between copies were detected within the 1541 bp long sequence. The impact of the sequence variability of 16S rRNA gene copies on phylogenetic positioning of P. bryantii was determined. All five sequences from cloned P. bryantii B(1)4 16S rRNA genes were placed in the same operational taxonomy unit. Control regions of all five analyzed rRNA operons were almost identical and three candidate for promoter sequences were identified by Neutral Network Promoter Prediction. Spacer regions between 16S rRNA and 23S rRNA genes in all five cloned copies were 543 bp long and genes for tRNA(Ile) and tRNA(Ala) were identified inside this regions. PMID- 11501482 TI - Restriction and modification systems of ruminal bacteria. AB - A high frequency of type II restriction endonuclease activities was detected in Selenomonas ruminantium but not in other rumen bacteria tested. Eight different restriction endonucleases were characterized in 17 strains coming from genetically homogeneous local population. Chromosomal DNA isolated from S. ruminantium strains was found to be refractory to cleavage by various restriction enzymes, implying the presence of methylase activities additional to those required for protection against the cellular endonucleases. The presence of Dam methylation was detected in S. ruminantium strains as well as in several other species belonging to the Sporomusa subbranch of low G + C Gram-positive bacteria (Megasphaera elsdenii, Mitsuokella multiacidus). PMID- 11501483 TI - Enrichment of bifidobacteria in the hen caeca by dietary inulin. AB - Caecal bifidobacterial concentration was increased more than 3-fold in inulin treated laying hens. The counts of bifidobacteria in birds fed as the control were 9.64, in inulin-diet fed ones 10.17 log CFU/g of caecal content, respectively. Dietary inulin had no effect on caecal microbial metabolite concentration. The proportion of inulin-fermenting bifidobacteria in the total bifidobacteria increased 2-fold in inulin-treated birds. PMID- 11501484 TI - Chitinolytic bacteria of the mammal digestive tract. AB - Chitinolytic bacteria were isolated from the digestive tract of different mammals and characterized. All isolates were facultatively anaerobic, long Gram-positive, straight rods resembling Clostridium sp. Only one isolate consisted of Gram positive ovoid cells. All cultures grew on glucose, N-acetylglucosamine, glucosamine, galactose, starch, hemicellulose and xylan. Fermentation products were mainly formate, acetate, butyrate and lactate. The isolates were identified as Clostridium sartagoforme (2 species), C. aminovalericum, C. bifermentans and Enterococcus durans (1 isolate of each species). Exocellular fractions of all strains exhibited higher activities of all enzymes than cellular ones. Inductive effects of hemicelluloses, pectin and laminarine on chitinases were demonstrated. High exocellular endochitinase activity was found in cultures grown on chitin. N Acetylglucosaminidase activity was low with the exception of exocellular fractions of two strains of C. sartagoforme. PMID- 11501485 TI - Isolation and characterization of rabbit caecal pectinolytic bacteria. AB - Two hundred and thirty colonies from the caecal contents of six rabbits were picked up and, after a 2-d incubation, were microscopically characterized using Gram staining. Large Gram-negative (34%) and small Gram-negative (30%) irregular rods, Gram-negative (27%) and Gram-positive (8%) cocci were found. Eleven isolates (Bacteroides ovatus (6 strains), B. thetaiotamicron, B. caccae, B. stercoris, B. capillosus and Capnocytophaga ochracea) were identified using commercial tests for measuring their catalase activity, metabolite production, etc., and testing their growth in 20% bile. Bacteria belonging to the genus Bacteroides were demonstrated to be the principal pectinolytic organisms in the rabbit caecum. PMID- 11501486 TI - Binding of extracellular matrix proteins by lactobacilli. AB - Ten gut and ten vaginal Lactobacillus strains were investigated for their ability to bind type I collagen (Cn-I) and four selected gut lactobacilli were investigated for their binding to other extracellular matrix (ECM) molecules. Immobilized Cn-I (100 mg/L) in wells of microtitre plates was bound by all 10 autoaggregating vaginal strains and by 3 strains of gut lactobacilli from piglets in the range of A570 readings 0.114-1.806. L. acidophilus strain SV31 was much more adherent than the rest of strains. All four gut lactobacilli tested for binding to other ECM molecules displayed good binding to porcine fibronectin and heparin and some of them bound weakly to fetuin and porcine mucin. No binding of these strains was observed to bovine mucin, bovine fibrinogen and bovine lactoferrin. PMID- 11501487 TI - Molecular monitoring of ruminal prevotellas. AB - The development and preliminary use of two different molecular approaches for rapid enumeration and monitoring of ruminal prevotellas are described. Several oligonucleotide DNA probes, specific for the genus Prevotella and the species P. ruminicola and P. bryantii were labeled with various fluorochromes and used in in situ hybridization experiments. Epifluorescent microscopy was successfully used for the detection of fluorescent signal emitted by the probes in pure and mixed culture samples. The enumeration of the target cells and the analysis of the crude rumen fluid proved to be difficult, however, mainly due to the autofluorescent background and nonhomogeneous distribution of the cells on the microscope slide. The development of a competitive PCR system for ruminal prevotellas is described and the preliminary results of the rumen fluid analysis from a black-and-white Holstein cow are given. PMID- 11501488 TI - Preliminary characterization of a tentatively novel rumen bacterial species from the genus Treponema. AB - A new spirochetal strain was isolated from the rumen of a black-and-white Holstein cow and preliminarily characterized. The sugar fermentation tests and morphological observations indicated this organism to be a member of a novel, as yet undescribed spirochetal rumen species. The small subunit ribosomal RNA genes were amplified and the PCR products were cut with the restriction endonucleases TaqI, DdeI, HhaI and Sau3AI. The comparison of the observed RFLP with the hypothetical fragment lengths of the computer analyzed 16S rRNA sequences from the type strains of the ruminal spirochetes Treponema bryantii and T. saccharophilum confirmed the tentative novel identification. Transmission electron microscopy showed that the bacterium has the typical spirochetal structures, i.e. the outer sheath, the protoplasmic cylinder and the axial filament (it is not yet clear how many flagella compose the filament). An additional extracellular structure was observed which appeared as an exocytoplasmic polar flagellum, approximately 2 microns long and protruding from one tip of the cell. The average size of the cells was 0.5 x 10-15 microns and the wavelengths and the amplitudes of the primary coils were 2.9 and 1.3 microns, respectively. PMID- 11501489 TI - Multiple endoxylanases of Butyrivibrio sp. AB - Butyrivibrio sp. Mz 5 with a high xylanolytic activity was isolated. Four major xylanases were detected in the cell-associated fraction using the zymogram technique. The xylanolytic activity was inducible with the oat spelts xylan; two endoxylanases (51 and 145 kDa) were formed constitutively. The bulk of the xylanolytic activity was cell-bound and growth-phase dependent; the maximum activity in the cell-associated fraction was achieved after 16 h of incubation. The highest xylanolytic activity was determined in a medium with 0.5% oat spelts xylan. Under optimum conditions (the highest xylanolytic activity produced), the two cell-bound xylanases (51 and 58 kDa) were isolated by anion exchange chromatography on CIM DEAE 8 tubes attached to a MPLC system, and gel filtration. PMID- 11501490 TI - The standard renal volume of Japanese boys and girls determined by three dimensional ultrasonography. AB - We calculated the volume of the kidney in 442 kidneys in 221 normal children (15 neonates, 45 infants, 82 boys and 79 girls between 1 and 15 years of age) by using three-dimensional ultrasonography. The standard curves of the renal volume by height and age were determined. No differences in the renal volumes were observed between boys and girls from 1 to 14 years of age, whereas the difference between volume in boys and girls 15 years of age was significant; the boys had larger kidneys. A significant correlation between height and the volume of the right kidney (r = 0.756, P < 0.0001), and between height and the volume of the left kidney (r = 0.791, P < 0.0001) was observed. In addition, a significant correlation was found between the renal volume and the standard renal weight in male and female Japanese children. The results provide a new standard for renal volume in Japanese children. It is considered that this standard curve might be useful in a clinical setting, as a reference for evaluating patients with renal diseases that may affect the volume of the kidneys. PMID- 11501491 TI - Angiogenic growth factors in patients with cyanotic congenital heart disease and in normal children. AB - Previous studies have demonstrated that the expression of angiogenic growth factors is induced in hypoxic models. However, little is known about these factors in patients with cyanotic heart disease. The purpose of this study was to examine the relationship between the plasma level of angiogenic growth factors and the severity of cyanosis. The study included 85 patients with cyanotic heart disease and age matched 81 controls. Median age was 4.2 years in the cyanotic group and 4.8 years in the control group. Mean systemic oxygen saturation was 80.6 +/- 7.3% in the cyanotic group and 98.1 +/- 0.5% in the control group. In the control group, vascular endothelial growth factor (VEGF) in the neonatal period was significantly elevated, then rapidly decreased within 3 months after birth. After 3 months of age, VEGF levels remained at a plateau. In contrast, this age dependency did not occur in hepatocyte growth factor (HGF) levels. Although VEGF and HGF levels were not different between the cyanotic and control groups within 3 months after birth, the VEGF level in the cyanotic group after 3 months of age was significantly elevated compared to the levels measured in the control group (149.2 +/- 105.6 vs. 66.3 +/- 22.5 pg/ml, p < 0.0001). Moreover, the VEGF level was negatively correlated with oxygen saturation (y = 440.6-3.53x, R = 0.47, p < 0.0001) in cases more than 3 months old. In contrast, no correlation was found between HGF level and oxygen saturation. Although physiologically increased VEGF in the neonatal period was rapidly decreased under normal oxygen saturation, a higher VEGF level persisted if systemic hypoxia was present. Persistently higher VEGF level may be related to the development of systemic to pulmonary collateral arteries in patients with cyanotic heart disease. PMID- 11501492 TI - Early diagnosis and treatment of pulmonary opportunistic infection by using polymerase chain reaction and beta-glucan in patients with hematological neoplasms. AB - The early diagnosis of 58 patients with hematological neoplasms accompanied by severe pulmonary infections of Pneumocystis carini (Pc), or Cytomegalovirus (CMV) pneumonia was made by polymerase chain reaction (PCR) using sputum samples, and of pulmonary mycosis by measuring blood beta-glucan levels by a Limulus test. The effectiveness of early treatment for opportunistic infection based on these early stage diagnosis was evaluated and the results of pathological analyses of the lung at autopsy were analyzed. PCR identified Pc pneumonia in 7 of the total 58 patients (12.1%), and early treatment was effective in all 7 patients (100%). PCR identified CMV pneumonia in 5 patients (8.6%), and early treatment was effective in 2 of the 5 patients. The level of beta-glucan confirmed mycotic pneumonia in 9 of these 58 patients (15.5%), and early treatment was effective in 7 of these (66.7%). These findings indicate that PCR and the beta-glucan method effectively enabled clinicians to diagnose pulmonary opportunistic infection in the early stage in 21 of the 58 patients (36.2%), and that early treatment was effective in 16 of the 21 patients (76.2%). The results of the pathological analyses of the lung at autopsy were: pulmonary tumor cell infiltration in a total of 5 patients (2 with ATL, 2 with NAE and 1 with AML); infection in a total of 6 patients (2 with ML and 4 with ATL); and diffused alveolar damage in a total of 4 patients (2 with ML, 1 with ATL and 1 with AML). CMV infection was confirmed in a total of 5 patients (2 with ML and 3 with ATL), and mucormycosis in a total of 2 patients (1 with ML and 1 with ATL). Despite these findings, Pc and other fungi or bacteria were not detected. Early diagnosis and treatment by the present PCR and beta glucan method were useful, but the underlying disease and its disease state influenced the clinical outcomes of patients with terminal pulmonary infection caused by CMV or mucor, suggesting that prevention and early diagnostic measures for these infections remain to be established. PMID- 11501493 TI - Quantification of WT1 mRNA by competitive NASBA in AML patients. AB - The measurement of Wilms' tumor gene (WT1) mRNA levels by reverse transcriptase polymerase chain reaction (RT-PCR) is useful in detecting minimal residual disease (MRD) in leukemia patients. In the present study, we quantified the level of WT1 mRNA in the peripheral blood and bone marrow of patients with acute myelocytic leukemia (AML) at initial onset, remission and recurrence by the use of nucleic acid sequence based amplification (NASBA), and then ascertained the clinical usefulness of this method. At initial onset, the level of WT1 mRNA in the peripheral blood was above 10(3) copies/microgram and that in the bone marrow was above 10(4) copies/microgram. The level of WT1 mRNA was decreased in cases where therapy resulted in complete remission, but it was abnormally high in recurring cases. In AML (M3) patients, the relationship between the level of WT1 mRNA and the expression of the PML-retinoic acid alpha receptor (RAR alpha) gene, assessed by fluorescence in situ hybridization (FISH), was investigated. When leukemia was in remission hematologically, the PML-RAR alpha gene was negative and the level of WT1 mRNA decreased. These findings suggest that the quantification of WT1 gene expression by competitive NASBA is useful in assessing therapeutic effects and detecting MRD. PMID- 11501494 TI - Soluble forms of P-, E- and L-selectin in children with Kawasaki disease. AB - Kawasaki disease (KD) is an acute, self-limiting systemic vasculitis syndrome of unknown origin, that mainly affects small and medium-sized arteries, particularly the coronary artery, which affects primarily infants and young children. Cell adhesion molecules play important roles in the inflammatory process. The aims of this study were to investigate the pathophysiological role of cell adhesion molecules in KD, and to look for the evidence of direct relationship between the plasma levels of soluble cell adhesion molecules and the incidence of coronary artery lesion (CAL). The 52 patients with KD, Group A patients who were clinical responders of initial intravenous immunoglobulin (IVIG) treatment (n = 30), Group B patients who did not respond to the initial IVIG treatment (n = 22), were studied. The circulating E-selectin (105.6 +/- 12.6 ng/ml) in the acute phase of KD, while the peak plasma P-selectin level (238.4 +/- 26.8 ng/ml) occurred in the subacute phase of illness (p < 0.05, respectively). Plasma L-selectin levels (1557.3 +/- 44.3 ng/ml) during the convalescent phase tend to higher than in the acute and in the convalescent phases (p = NS). The analysis of paired samples in Group A patients before (E-selectin: 131.2 +/- 9.8 ng/ml, P-selectin: 216.6 +/- 13.4 ng/ml) and 48 hour after (E-selectin: 98.9 +/- 9.2 ng/ml, P-selectin: 153.9 +/- 34.1 ng/ml) IVIG administration revealed significantly lower values of E- and P-selectins, however, no significant differences in those in Group B patients. There were also no significant differences in the values of L-selectins between the 2 groups. Before IVIG treatment, the plasma levels of E- (225.1 +/- 46.1 ng/ml) and P-selectin (259.4 +/- 76.2 ng/ml) of patients with CAL (n = 11) were significantly higher than those of patients (n = 41) without CAL (p < 0.05, respectively). Plasma L-selectin levels (1596.9 +/- 385.0 ng/ml) in patients with CAL tended to be lower than those in patients without CAL (p = NS). E- and P selectin may have potential as predictors of CAL in patients with KD. PMID- 11501495 TI - Acid-base imbalance adapts without changes in cell polarity in cortical collecting ducts in premature rabbits. AB - It has been demonstrated that intercalated cells (ICs) change their cellular composition in acid stimuli in adult rabbits but not fully explained in developing rabbits. To clarify the mechanisms of adaptation in acid-base imbalance in collecting ducts in developing rabbit kidneys, we examined the cellular composition of cortical collecting ducts (CCDs) in 4-week-old rabbits. In the control group, the ratio of ICs to total CCD cells and that of peanut agglutinin (PNA) non-binding ICs to total ICs were 37.2 +/- 7.2% and 40.3 +/- 3.1%, respectively. By contrast, inconsistent with adult reports, in the acidotic group, these ratios were 38.4 +/- 5.1% and 41.9 +/- 1.7%, respectively, similar to the control group. The urinary pH in the control group was 8.20 +/- 0.14, while that in acidemia was 4.98 +/- 0.33 (p < 0.01). These data indicated that cellular remodeling of ICs in the acidotic state is less important for adaptation in the 4-week-old rabbit. Another mechanism, employing an acid-base related protein, might be playing an important role during development for acid base imbalance. PMID- 11501496 TI - Cryopreservation and engraftment potential of peripheral blood stem cells: pediatric experience. AB - Peripheral blood stem cells (PBSC) harvests were performed from children with various types of cancer in our institution. The PBSC was cryopreserved using the simplified method at -80 degrees C with 6% hydroxyethyl starch (HES) and 5% dimethyl sulfoxide (DMSO) without a programmed freezer. To determine the optimum cryo-storage time in this method, 33 samples harvested from 33 patients were investigated. While there was no correlation between the time of cryopreservation and the nucleated cell recovery rates, the CFU-GM recovery rate was negatively correlated with the cryopreservation times (p < 0.01). The CFU-GM recovery rate after 2, 3, and 5 years cryopreservation was 74%, 56%, and 21%, respectively. To date, 22 of the 33 children have subsequently received PBSCT using their cryopreserved cells. The number of infused CFU-GM of PBSCs was 6.9-114.6 (median 20.5) x 10(4)/kg. The cryopreservation time of infused PBSCs was 1-35 (median 4) months. After PBSCT, the absolute neutrophils count (ANC) achieved 500/mm3 between 8-16 (median 10.5) days, and the platelet count achieved 5.0 x 10(4)/mm3 between 13-200 (median 29) days. We have experienced PBSCT using PBSCs after 35 months cryopreservation as the maximum and successful engraftment. It has been clarified in this study that PBSCs could be clinically used successfully after 3 years cryopreservation. PMID- 11501497 TI - Detection of the retinoic acid-regulated genes in a RTBM1 neuroblastoma cell line using cDNA microarray. AB - A microarray system is a powerful and very useful technology for analyzing the expression profile of thousands of genes. In this study, we made a cDNA microarray system carrying 2007 cDNAs obtained from primary neuroblastoma cDNA library and identified retinoic acid (RA)-regulated genes in a RTBM1 neuroblastoma cell line. We repeated independent hybridization experiment twice and found that 7 genes were up-regulated, and 5 genes were down-regulated on the cDNA microarray. The semi-quantitative reverse transcriptase (RT)-PCR analysis to confirm the results showed that 4 genes which included amyloid precursor-like protein 2 (APLP2), P311, dihydropyrimidinase related protein3 (DRP3) and RGP4 were up-regulated, while 2 genes, Id-2 and vimentin, were down-regulated. Thus, our neuroblastoma cDNA microarray system is useful to screen the neuronal differentiation- and growth-related genes regulated by RA with high efficiency. PMID- 11501498 TI - Prevalence of Chlamydia pneumoniae in acute respiratory tract infection and detection of anti-Chlamydia pneumoniae-specific IgE in Japanese children with reactive airway disease. AB - We examined the prevalence of Chlamydia pneumoniae in acute respiratory tract infection and association of C. pneumoniae infection and reactive airway disease in Japanese children. Four hundred eleven children with acute respiratory tract infection were enrolled in this study, and C. pneumoniae was isolated from 58 (14.1%) patients by culture. Evidence of infection with C. pneumoniae was detected in 58 children with pneumonia (34.5%), bronchitis (41.4%) and upper respiratory tract infection (24.1%). Twenty-nine (50.0%) out of 58 patients were younger than 5 years old and 18 (31.0%) had wheezing at first visit. A logistic test for anti-C. pneumoniae-specific IgE showed the deference in the fluorescence unit between the patients with C. pneumoniae infection with and without wheezing was statistically significant (Po = 0.02748, to = 2.31891). In conclusion, C. pneumoniae seems to be an important respiratory tract pathogen among young Japanese children, and our results support the association of C. pneumoniae infection and reactive airway disease. PMID- 11501499 TI - Interferon-alpha: an effective adjuvant for peptide-based cytotoxic T-cell vaccines. AB - An important issue for developing a vaccine therapy for human malignancy is identifying adjuvants that can elicit T-cell responses to peptides. The present study evaluates interferon-alpha (IFN-alpha) as a vaccine adjuvant. C57BL/6 mice were immunized subcutaneously with peptide derived from influenza virus (Flu) either with or without IFN-alpha using different vaccine formulations. IFN-alpha significantly enhanced cytotoxic T lymphocytes (CTL) induction in mice immunized with Flu peptide in incomplete Freund's adjuvant (IFA). Flu peptide administered continuously for 3 days by osmotic pump with IFN-alpha could elicit CTL induction, whereas either Flu peptide or IFN-alpha alone was non immunogenic. Furthermore, injection of the liquid formation of Flu peptide with IFN-alpha in phosphate-buffered saline (PBS) did not elicit CTL induction. These results suggest that the continuous administration of peptide and local delivery of IFN alpha are important for efficient CTL induction, and that IFN-alpha is an effective adjuvant for peptide-based vaccines. PMID- 11501500 TI - Role of gamma/delta T-cells in the peripheral blood of patients with pulmonary tuberculosis. AB - Although gamma/delta T-cells are known to contain the highest frequency of mycobacteria-reactive cells in humans, and recent studies have suggested that they play an important role in the initial immune response to Mycobacterium tuberculosis (Mtb), very few studies have attempted to analyze these cells in patients with active pulmonary tuberculosis (TB). The aim of the present study was therefore to evaluate the gamma/delta T-cell populations present in the peripheral blood and the IFN-gamma production of gamma/delta T-cells stimulated by PMA before and after anti-TB chemotherapy in patients in the initial treatment stage for primary active pulmonary TB. Cell populations were measured by three color flow cytometry of peripheral blood mononuclear cells. We compared the population of gamma/delta T-cells and the production of IFN-gamma between normal healthy controls and TB patients. Absolute numbers of gamma/delta T-cells remained constant in the peripheral blood of TB patients. However, production of IFN-gamma in gamma/delta T-cells was dramatically suppressed prior to anti-TB chemotherapy in comparison with healthy control subjects, and further reduced following anti-TB chemotherapy. We also examined the influence of isoniazid (INH) in anti-TB chemotherapy. INH suppressed IFN-gamma production of gamma/delta and alpha/beta T-cells. The findings demonstrated a strong correlation between the production of IFN-gamma in gamma/delta T-cells and manifestation of primary active pulmonary TB, which was consistent with the hypothesized role for gamma/delta T-cells in the protective immune response to Mtb infection. PMID- 11501501 TI - A case of ileus caused by implantation of cancer after surgical treatment of bile duct carcinoma. AB - We encountered a patient with an ileus caused by implantation of cancer cells after surgical treatment of bile duct carcinoma. The patient was a 55-year-old male diagnosed as having cancer in the lower bile duct who underwent pancreatoduodenectomy in March 1996. The comprehensive curability was B. The patient experienced nausea and vomiting and was diagnosed as having ileus in October 1998. The symptoms were alleviated by medical treatment. Because ileus symptoms recurred in December 1998 but were not alleviated by medical treatment, the patient was hospitalized in Kurume University Hospital for surgery. A mirror reflection was observed by plain radiography of the abdomen. Ultrasonography (US) revealed dilation of the intestinal duct and hypertrophy of the intestinal wall. An ileus tube was inserted but the symptoms did not improve, and therefore, the patient underwent surgery in February 1999. Upon laparotomy, a node of milk white color about 2 cm in size was found in the region coinciding with the enhancement on CT images, and part of the intestinal duct was found to be folded and adhered to the lower surface of the liver. No recurrence was observed in the liver, peritoneum, and lymph nodes. The tumor was excised together with part of the intestinal duct and liver to remove the ileus. At present, the patient is alive without recurrence. Because recurrence of cancer by implantation of cancer cells is often localized in a limited region, early diagnosis and excision are important. PMID- 11501502 TI - Adult colonic intussusception: a case report. AB - Intussusception accounts for almost all cases of intestinal obstruction in children. In contrast, intussusception in adults is relatively rare. An 86-year old Japanese female with rectal bleeding came to our hospital via ambulance. At first, colonoscopy findings revealed the sigmoid colon cancer. Ultrasonography showed a hypoechoic mass with a multiple concentric ring sign. Computed tomography showed a round fluid-filled cystic structure. Colon contrast studies demonstrated stenosis in the rectosigmoid colon. A laparotomy was performed. The sigmoid colon was intussuscepted to the rectosigmoid colon. We employed both rectosigmoid and sigmoid colon resection. The resected specimen showed that the disease was advanced sigmoid colon cancer with ulcer formation due to an ischemic change. Tumor was 4.5 cm x 2.0 cm in size. The disease was histopathologically diagnosed as advanced sigmoid colon cancer, well-differentiated adenocarcinoma. We report here a case of adult intussusception due to the sigmoid colon cancer. PMID- 11501503 TI - Placental transport of bile acids: analysis of bile acids in maternal serum and urine, umbilical cord blood, and amniotic fluid. AB - To investigate the role of placental transport of bile acids in fetal bile acid metabolism, such as with regard to synthesis of the unusual bile acids (1 beta- and 6 alpha-hydroxylated and unsaturated bile acids), we measured the concentrations of bile acids in umbilical cord blood, amniotic fluid, maternal serum and maternal urine at delivery by means of gas chromatography-mass spectrometry. Serum and urine from healthy nonpregnant women were used as controls. We detected large amounts of unusual bile acids, especially hyocholic acid and 3 beta-hydroxy-delta 5 bile acids, in amniotic fluid and umbilical cord blood. The concentration of total bile acids in maternal serum was less than that of control serum and umbilical cord blood, and the concentration of total bile acids in maternal urine was higher than that of control urine and amniotic fluid. In conclusion, the fetus synthesized large amounts of unusual bile acids, and these compounds were transported from fetus to mother by placental transfer. We suggest that pregnant women may excrete large amounts of bile acids into the urine to control serum concentration of bile acids in fetus. PMID- 11501504 TI - Dihydropyrimidine dehydrogenase activity and thymidylate synthase level are associated with response to 5-fluorouracil in human colorectal cancer. AB - In the recent studies associated with the modulation of 5-fluorouracil (5-FU) and the development of new antifolates, attentions have been focused on the expression of the target enzymes, thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD), that affect tumor sensitivity and resistance to drugs. In order to evaluate predictability of therapeutic efficacy by intratumoral enzyme activity, we investigated the role of TS content and DPD activity in tumor sensitivity of 5-FU. Surgical specimens were obtained from 51 patients with colorectal cancer and used to measure TS content and DPD activity. TS content and DPD activity in tissues were measured by [3H]-FdUMP binding assay and radioenzymatic assay, respectively. The sensitivity to 5-FU in tumor specimens was determined by collagen-gel droplet embedded-drug sensitivity test (CD-DST). The TS content and DPD activity did not correlate with Dukes' staging. There was no correlation between TS content and DPD activity in any tumors. Simple linear regression analysis showed that neither DPD activity (r = -0.267, p > 0.05) nor TS content (r = -0.277, p > 0.05) in tumors had a significant correlation with 5 FU effectiveness independently. Four out of 24 patients, highly responsive to 5 FU, showed low levels in both DPD and TS. The patients with high value in either DPD activity or TS content proved not to respond to 5-FU. In conclusion, these results demonstrate that both tumor DPD activity and TS content are the factors predicting 5-FU responsiveness in colorectal cancer. PMID- 11501505 TI - The usefulness of restorative laparoscopic-assisted total colectomy for ulcerative colitis. AB - The aim of this study was to evaluate the effectiveness of laparoscopic-assisted total colectomy with ileal J-pouch anal anastomosis in comparison with that of conventional open total colectomy in patients with ulcerative colitis. From 1990 until 1999, 11 patients underwent open total colectomy, while 21 patients had laparoscopic-assisted total colectomy. Patients characteristics, perioperative course, and recovery were compared. Differences between the groups were tested using Student's t-test for independent groups and chi-square tests when appropriate. Nasogastric tube could be removed after POD 1.1 vs. 4.8 (p < 0.05), the mean time to passage of stool was 1.7 (range, 1 to 3) vs. 5.4 (range, 3 to 7) days (p < 0.05), and in the laparoscopic group watery stool was soon made solid after POD 24.3 vs. 87.3 (p < 0.01). There were no significant differences in the mean operating time and mean estimated operating blood loss between two groups. Postoperative morbidity did not differ significantly between the patients treated conventionally (45.5%) and laparoscopically (33.3%). The findings of this study indicate that laparoscopic-assisted total colectomy improved cosmetic results and to be a safe and effective treatment for the elective surgery of ulcerative colitis. PMID- 11501506 TI - Proceedings of the 3rd International Symposium on Growth Hormone Secretagogues. Keystone, Colorado, USA. February 2001. PMID- 11501507 TI - Efficacy and tolerance of topical calcitriol 3 microg g(-1) in psoriasis treatment: a review of our experience in Poland. AB - Although topical vitamin D3 derivatives have been used in the treatment of patients with psoriasis for the past 15 years, questions remain about the indications and limitations of application. Extensive personal experience gained during the development of calcitriol (1alpha25-dihydroxyvitamin D3) is therefore reviewed. Three double-blind, vehicle-controlled trials have revealed that calcitriol 3 microg g(-1) ointment (Silkis ointment, Galderma Laboratories) has very good clinical efficacy. In a left-right comparison with vehicle ointment, complete clearance of psoriatic lesions was achieved in 48% of sites treated with calcitriol and a further 41% showed considerable or definite improvement. The clinical response to calcitriol in another study was as good as, or even better than, that achieved with betamethasone valerate 0.1% ointment. A preparation containing calcitriol 15 microg g(-1) did not show any clinical superiority to the lower dose but was associated with a higher risk of hypercalciuria, particularly when applied to extensive skin lesions. These results suggest that calcitriol 3 microg g(-1) ointment is an effective and safe treatment for chronic plaque psoriasis. PMID- 11501508 TI - Long-term safety of topical calcitriol 3 microg g(-1) ointment. AB - To assess the topical and systemic safety and tolerance of twice-daily application of 3 microg g(-1) 1alpha25-dihydroxyvitamin D3 (calcitriol) ointment (Silkis ointment, Galderma Laboratories) in the long-term treatment of patients suffering from chronic plaque psoriasis, we performed an open-design, multicentre study. Two hundred and fifty-three patients (155 males, 98 females) treated all their psoriatic lesions, except for those on the head and scalp, for up to 78 weeks. No serious adverse events were reported: 37 patients (14.6%) had a transient skin irritation reaction on one or more occasions during the study that resulted in study withdrawal for seven of them. The baseline/endpoint analyses showed no clinically relevant changes in measures of calcium and phosphorus homeostasis and renal function. Slight hypercalcaemia was observed in five (2%) patients: in four of these patients, serum albumin-adjusted total calcium levels normalized during treatment. In conclusion, twice-daily calcitriol 3 microg g(-1) ointment is safe and well tolerated in the long-term treatment of chronic plaque psoriasis. PMID- 11501509 TI - Clinical experience with topical calcitriol (1,25-dihydroxyvitamin D3) in psoriasis. AB - The use of vitamin D analogues for the treatment of chronic plaque psoriasis is well documented. Of importance now is their comparability and compatibility with other established treatments for psoriasis. This paper reviews five studies with calcitriol 3 microg g(-1) ointment (Silkis ointment, Galderma Laboratories). Calcitriol applied twice daily was found to be as effective as short-contact dithranol in terms of global improvement and PASI scores. However, patients favoured calcitriol over dithranol when both quality of life and treatment acceptability were assessed. Two studies provide evidence of the benefit of combining calcitriol with other antipsoriatic therapies. Combination with ultraviolet (UV) B phototherapy proved as effective as UVB alone over an 8-week period; however, the combination had a radiation dose-sparing effect, thus reducing the risk of adverse events. Likewise, calcitriol combined with betamethasone valerate (each applied separately, once daily) was as efficacious as twice-daily betamethasone, thereby achieving a corticosteroid-sparing effect. Finally, two studies confirm that calcitriol 3 microg g(-1) ointment can be used safely in patients with psoriasis of the head and confirm the high level of clinical efficacy achieved with this compound. PMID- 11501510 TI - Combination therapy with vitamin D analogues. AB - Monotherapy with vitamin D analogues has been shown to be effective in the treatment of psoriasis. Vitamin D analogues have also been used in combination with other topical therapies, systemic therapies and phototherapy. In many instances, the efficacy of these other treatments can be maximized and adverse effects minimized when combined with vitamin D analogues. The combination of a topical corticosteroid with a vitamin D analogue can work synergistically to improve efficacy and reduce the side-effects from both treatments. However, caution must be used when mixing the two agents, as some topical corticosteroids will result in the degradation of the vitamin D analogue. Benefit from phototherapy is also increased when using vitamin D analogues, so that greater improvement occurs with fewer treatments. Effects on minimal erythema dose must be considered and the potential for ultraviolet blocking by vitamin D analogues may affect treatment. Some vitamin D analogues may also be susceptible to degradation by certain wavelengths of ultraviolet light. Combining vitamin D analogues with systemic agents exerts a dose-sparing effect, thus reducing the possibility of side-effects, but such combinations require further study. As long as treatments are used correctly, the benefits of combination therapy with vitamin D analogues usually outweigh the few drawbacks. PMID- 11501511 TI - Topical calcitriol--studies on local tolerance and systemic safety. AB - Calcitriol 3 microg g(-1) ointment (Silkis ointment, Galderma Laboratories) is a new treatment for psoriasis. Calcitriol is the biologically active metabolite of vitamin D3. It induces keratinocyte differentiation, inhibits keratinocyte, T cell and fibroblast proliferation, and inhibits the production of some inflammatory mediators, all contributors to the pathogenesis of psoriasis. Preclinical studies have shown an effect of topical calcitriol on calcium homeostasis at doses higher than those in clinical use. No adverse local events were observed when calcitriol was applied to animal skin. Phase I clinical studies confirmed that calcitriol 3 microg g(-1) ointment is well tolerated in humans. These studies have demonstrated that at the minimal effective concentration of 3 microg g(-1), calcitriol ointment has no discernible photosensitizing or phototoxic potential and no skin irritant or allergic potential in healthy volunteers. Its low systemic absorption through human skin is unlikely to significantly affect calcium homeostasis. This paper summarizes the findings of the preclinical and early clinical studies that provided the foundation of the later Phase II and III clinical trials on efficacy and safety with topical calcitriol 3 microg g(-1) ointment for the treatment of plaque psoriasis. PMID- 11501512 TI - The impact of psoriasis on quality of life. AB - Psoriasis is a chronic inflammatory skin disease in which the signs vary from one patient to another and over time. Traditionally, physicians have used various parameters to assess the severity of the disease: percentage of body surface area covered, erythema, plaque thickness, degree of scaling and systemic symptoms such as arthritis. However, these clinical assessments alone do not accurately reflect the overall effect of the disease on patients' daily activities. Apart from the clinical severity of affected areas, psoriasis can also have a profound psychosocial impact on the patient's quality of life. This concept is multidimensional, encompassing the physical, social and psychological wellbeing of the person and is based on the patient's view of their condition. PMID- 11501513 TI - Psoriasis: the future. AB - The aetiology of psoriasis is still unclear but our knowledge of the psoriatic process has grown substantially over the last two decades. The future will undoubtedly bring advances in our understanding of the pathogenesis of psoriasis and, as a consequence, new therapies. Defining the molecular genetics of psoriasis will enhance our understanding of the disease process and hopefully facilitate the development of a representative animal model. This in itself will be a key step in the development and testing of new therapies. Precise identification of the immunological events involved in psoriasis will allow specific T-cell- and cytokine-targeted, and perhaps less toxic. immunotherapies. Anti-angiogenic agents that are in development for use in oncology may also be effective in psoriasis. The adaptation of current topical therapies such as retinoids and vitamin D analogues to produce more effective and better-tolerated formulations will also play a significant role in our future first-line management of patients. The increased recognition and better management of environmental trigger factors such as psychological distress will become an important factor in future psoriasis care. The development of physical therapies including photodynamic therapy and excimer lasers has the potential to expand the remit of psoriasis therapy. There is little doubt that the future for our patients with psoriasis is bright. However, this will only be achievable by a concerted research effort to understand all facets of this enigmatic disease ranging from the molecular to the environmental. PMID- 11501514 TI - Fluoride--Seen from Different Perspectives. Proceedings of a workshop. Amsterdam, The Netherlands, November 4, 2000. PMID- 11501516 TI - Proceedings of the Symposium on Immunological, Chemical and Clinical Problems of Food Allergy. 11-13 March 2001, Venice, Italy. PMID- 11501515 TI - Life-threatening giant mediastinal goiter: a surgical challenge. AB - Mediastinal goiter is a well known benign disease, usually resectable through a cervical approach with minimal morbidity and mortality. Only occasionally a median sternotomy or a lateral thoracotomy may be required. The present case is worthy of presentation because of the exceptional dimension of the disease and the surgical challenge that it presented. In a 72-year-old woman a large intrathoracic goiter of the right thorax caused a severe dyspnoea due to an important contralateral mediastinal shift with compression of the lung, superior vena cava system and trachea. At surgical exploration, through a cervico sternotomic approach, the mediastinal structures dislocation and the strong adherences between the anomalous neovascularized capsula of the mass and the surrounding structures, complicated the surgical dissection. An accidental lesion of the innominate artery required its reimplantation on the ascending aorta. An immense mass, was finally removed and pathological examination revealed a rare case of neovascularized, pseudosarcomatoid capsula among a benign hyperplastic proliferation. In spite of its benign nature, a giant goiter caused a life threatening compression of the respiratory tract and lung parenchyma in this patient. The dimension of the lesions, the mediastinal anatomy alterations and the severe intraoperative haemorrhage represented major technical difficulties during surgical resection. PMID- 11501517 TI - A commitment to safety. A toolkit for JCAHO's new patient safety standards. PMID- 11501518 TI - Visible light-curing: part I. PMID- 11501519 TI - Familial uveal melanoma: report on three sibling cases. PMID- 11501520 TI - Herpes simplex dendritic keratitis after treatment with latanoprost for primary open angle glaucoma. PMID- 11501521 TI - Optic neuropathy and cerebellar ataxia associated with a rare missense variation (A14510G) of mitochondrial DNA. PMID- 11501522 TI - The tuberculointerstitial nephritis and uveitis (TINU) syndrome is associated with HLA-DR14 in Spanish patients. PMID- 11501523 TI - Idiopathic polypoidal choroidal vasculopathy in a patient with atrophic age related macular degeneration. PMID- 11501524 TI - Spontaneous orbital haemorrhage following cardiac angioplasty. PMID- 11501525 TI - Eyes in the back of your head. PMID- 11501526 TI - [2nd Symposium on Evidence Based Medicine. 5-7 October 2000, Berlin, Germany. Abstracts]. PMID- 11501527 TI - Ligand binding regions in the receptor for urokinase-type plasminogen activator. AB - The interaction between urokinase plasminogen activator (uPA) and its cellular receptor (uPAR) is a key event in cell surface-associated plasminogen activation, relevant for cell migration and invasion. In order to define receptor recognition sites for uPA, we have expressed uPAR fragments as fusion products with the minor coat protein on the surface of M13 bacteriophages. Sequence analysis of cDNA fragments encoding uPA-binding peptides indicated the existence of a composite uPA-binding structure including all three uPAR domains. This finding was confirmed by experiments using an overlapping 15-mer peptide array covering the entire uPAR molecule. Four regions within the uPAR sequence were found to directly bind to uPA: two distinct regions containing amino acids 13--20 and amino acids 74--84 of the uPAR domain I, and regions in the putative loop 3 of the domains II and III. All the uPA-binding fragments from the three domains were shown to have an agonistic effect on uPA binding to immobilized uPAR. Furthermore, uPAR-(154--176) increased uPAR-transfected BAF3-cell adhesion on vitronectin in the presence of uPA, whereas uPAR-(247--276) stimulated the cell adhesion both in the absence or presence of uPA. The latter fragment was also able to augment the binding of vitronectin to uPAR in a purified system, thereby mimicking the effect of uPA on this interaction. These results indicate that uPA binding can take place to particular part(s) on several uPAR molecules and that direct uPAR-uPAR contacts may contribute to receptor activation and ligand binding. PMID- 11501528 TI - Measurement of atherosclerosis progression. PMID- 11501529 TI - Measurement of atherosclerosis progression. PMID- 11501530 TI - Measurement of atherosclerosis progression. PMID- 11501531 TI - Smoking and wrinkling of the skin. PMID- 11501532 TI - Serum soluble interleukin-2-receptor concentrations. PMID- 11501533 TI - Relevance of high and low penetrance. PMID- 11501534 TI - Gnathostomiasis. PMID- 11501535 TI - Cautions over idiopathic aldosteronism. PMID- 11501536 TI - Cautions over idiopathic aldosteronism. PMID- 11501537 TI - Cautions over idiopathic aldosteronism. PMID- 11501538 TI - Reye's syndrome. PMID- 11501539 TI - US drug policy. PMID- 11501540 TI - Proposed definitions for terminal sedation. PMID- 11501543 TI - Abstracts of the British Geriatrics Society Spring Meeting. 5-7 April 2001, Cardiff, United Kingdom. PMID- 11501542 TI - Abstracts of the British Association of Dermatologists 81st Annual Meeting. 3-7 July 2001. Cardiff, United Kingdom. PMID- 11501544 TI - [III National Convention on the Global Approach to the Cancer Patient, from Innovative Treatment to Palliative Cure. Rimini, Italy, 6-8 June 2001. Abstracts]. PMID- 11501545 TI - Abstracts of the International Meeting on Inflammatory Bowel Diseases. Capri, Italy, June 18-21, 2000. PMID- 11501546 TI - Bacteroides fragilis arthritis in a patient with sickle cell disease. PMID- 11501547 TI - What role would thiazide diuretics have in the prevention of postmenopausal osteoporosis? PMID- 11501548 TI - Acute oliguric renal failure associated with angiotensin II receptor antagonists. PMID- 11501549 TI - Disappearance of antiphospholipid antibodies syndrome after Helicobacter pylori eradication. PMID- 11501550 TI - Gemella morbillorum bacteremia associated with adenocarcinoma of the cecum. PMID- 11501551 TI - Eleven-year survival of a patient with bullous sarcoidosis after bilateral pleurodesis. PMID- 11501552 TI - Increased plasma adrenomedullin levels in Kawasaki disease with coronary artery involvement. PMID- 11501553 TI - Vertebral osteomyelitis due to Corynebacterium striatum. PMID- 11501555 TI - Abstracts of the 27th Annual Meeting of the European Thyroid Association. Warsaw, Poland, August 25-29, 2001. PMID- 11501554 TI - Pulmonary hyperinfection syndrome with Strongyloides stercoralis. PMID- 11501556 TI - Canadian Surgery Forum. Quebec, Canada, September 6-9, 2001. Abstracts. PMID- 11501557 TI - EPICOH 2001. 15th Symposium on Epidemiology in Occupational Health. Copenhagen, Denmark, 20-22 August 2001. Abstracts. PMID- 11501558 TI - EUROMEDLAB 2001. Abstracts of the 14th IFCC-FESCC European Congress of Clinical Chemistry and Laboratory Medicine, 5th Czech National Congress of Clinical Biochemistry. Prague, Czech Republic, May 26-31, 2001. PMID- 11501559 TI - Abstracts of the 2nd ASI (Arbeitsgemeinschaft Stabile Isotope) Meeting. October 4 6, 2000. Leipzig, Germany. PMID- 11501560 TI - Abstracts of the 10th International Congress on Cardiovascular Pharmacotherapy. Kyoto, Japan, 27-31 March 2001. PMID- 11501561 TI - A Decade of ELSI Research: A Celebration of the first ten years of the Ethical, Legal, and Social Implications (ELSI) Programs. January 16-18, 2001. Abstracts. PMID- 11501562 TI - Length measurement: new developments in neurostereology and 3D imagery. AB - Quantification of linear biological structures has important applications in neuroscience; for example, the length of neurotransmitter-specific axonal innervation or length of dendritic processes within particular brain structures. Until recently, however, there have been practical limitations in the application of stereological tools for the unbiased estimation of object length on tissue sections. The recent development of efficient new approaches allows for the wider application of theoretically unbiased sampling and estimation techniques that are devoid of the assumptions and models of earlier methods. In this review, we outline the historical background and recent advances in the estimation of total length for biological objects on tissue sections, including a practical method to estimate the length of cholinergic fibers using newly developed methods. These newer methods also take advantage of three-dimensional image datasets and virtual probes, techniques that may have wider application in quantitative morphometry. PMID- 11501563 TI - Redefined nomenclature for members of the carcinoembryonic antigen family. PMID- 11501564 TI - ACOG Committee Opinion. Committee on Ethics. Human immunodeficiency virus: Ethical guidelines for obstetricians and gynecologists. PMID- 11501566 TI - ACOG Committee Opinion no. 256: Committee on Obstetric Practice. Optimal goals for anesthesia care in obstetrics. PMID- 11501565 TI - ACOG Practice Bulletin. Clinical management guidelines for obstetrician gynecologists. Medical management of abortion. AB - According to the U.S. Centers for Disease Control and Prevention, 1.18 million legal abortions were performed in the United States in 1997. Of these, 55.5% were performed at or before 56 days of gestation (calculated from the first day of the last menstrual period [LMP]) (I). Almost 98% of abortion procedures were performed by uterine curettage; all but 1% of these used suction curettage. There were 305 legal induced abortions per 1,000 live births, and the abortion rate was 20 per 1,000 women aged 15-44 years. For the first time in 1997, medical abortions were counted and comprised 0.25% of all abortions; 0.45% of those procedures were performed up to 56 days of gestation. Because of the lack of availability of mifepristone, these procedures mostly represent the use of a combination of methotrexate and misoprostol. Over the past two decades, medical methods of abortion have developed throughout the world and are now used clinically in the United States. This document will present evidence of effectiveness, benefits, and risks of medical methods of abortion and provide a framework for the evaluation and counseling of women who are considering such medical methods. PMID- 11501567 TI - ACOG Practice Bulletin No. 27: Clinical Management Guidelines for Obstetrician Gynecologists. Prenatal diagnosis of fetal chromosomal abnormalities. AB - The prevalence of chromosomal abnormalities in clinically recognized early pregnancy loss is approximately 50% (1). Aneuploid fetuses account for 6-11% of all stillbirths and neonatal deaths (2). Chromosome defects compatible with life but causing significant morbidity occur in 0.65% of newborns, and another 0.2% have structural chromosomal rearrangements that will eventually affect reproduction (3). Although it is not possible to identify all aneuploidies antenatally, screening and diagnostic programs to detect the most common autosomal trisomy in liveborn infants, Down syndrome, are well established. This document will provide clinical management guidelines for the prenatal detection of these aneuploidies. PMID- 11501568 TI - ACOG Practice Bulletin no. 28: Clinical Management Guidelines for Obstetrician Gynecologists. Use of botanicals for management of menopausal symptoms. AB - Lack of confidence in the espoused benefits of hormone replacement therapy (HRT) coupled with a significant array of side effects of HRT, results in fewer than 1 in 3 women choosing to take HRT. The use of alternatives to conventional HRT has become more accessible and acceptable to many women. As more women choose these alternatives, physicians are confronted with the challenges of how to advise patients about alternative medicine and how to determine which therapies may be safe and effective. This document will examine available scientific information on alternative therapies for treatment of menopausal symptoms and provide recommendations on efficacy and potential adverse consequences. PMID- 11501569 TI - Safety and efficacy of AS-1 red blood cell use in neonates. AB - Many Regional Blood Centers are providing AS-1(Adsol preservative) red blood cells (RBCs) as a standard product because of the extended shelf life (42 days). The use of AS-1 RBCs is concerning in neonates because of high exposure to dextrose, adenine and mannitol. We conducted this study to evaluate the safety and efficacy of AS-1 RBC use in neonates. We assigned one unit of AS-1 RBCs to each infant for small volume transfusions (15 ml/kg) for the life of the unit (42 days). The study was conducted for one year. The infants under 1500 g were included in the study. We measured the pre- and post-transfusion hematocrit, post transfusion serum sodium, potassium, glucose, bilirubin and blood pH. We compared the average number of transfusions per patient and average blood donor exposure per patient using AS-1 RBC to CPDA-1 packed red blood cells (PRBC) use, data available for prior year. We monitored the blood transfusion reactions during the study period. The hematocrit increased significantly from 30.1 +/- 4.6 pre transfusion to 38.3 +/- 4.9 post-transfusion. The post-transfusion serum bilirubin, blood pH, serum potassium, sodium and glucose remained within the normal range. In spite of an increase in the number of average transfusions per patient with AS-1 RBC (6.67 +/- 5.1), the average donor exposure (1.8 +/- 1.1) remained less than two donors. There were not any transfusion reactions reported during the study. In conclusion, the use of AS-1 red blood cells is safe for small volume transfusions in neonates. PMID- 11501570 TI - Generation of dendritic cells: role of cytokines and potential clinical applications. AB - Over the last decade there has been great interest in generating populations of antigen presenting cells (APC), which can be exploited to improve immune responses to a variety of diseases including malignancies. Dendritic cells (DC) are an APC population that are easily generated ex vivo using a variety of cytokines. Cytokines can also be used to further manipulate these cells in maturation and function. The following discussion will provide an overview of dendritic cell isolation and generation with a focus on the role cytokines play in this process. PMID- 11501571 TI - First experience with a ready-for-use rheohemapheresis system. AB - Rheohemapheresis is increasingly being used for the improvement of microcirculation in numerous diseases. The method is based on the constant-flow separation of plasma by a cell separator and the secondary filtration of plasma through a hollow-fiber membrane. A new CE-marked system has recently been launched for an improved continuous flow blood separator (dideco Excel Pro) that contains a connection kit between the cell separator and a secondary filter and software which includes differential filtration as a standard protocol. The new system was used in our center using ethylenevinylalcohol secondary filters (Evaflux LA4 or LA5). 99 procedures were completed in 47 patients. A median of 2.7 (1.6-4.2) 1 of plasma were processed via the secondary filter in 109 (41-218) min. The values in peripheral blood before and after the treatment were total protein 6.7 (5.8-8.9)/ 5.3 (4.4-6.7) g/dl, fibrinogen 215 (118-480)/110 (37-275) mg/dl and cholesterol 200 (134-254)/92 (69 149) mg/dl. A median platelet loss of 30% in the peripheral blood of the patients was observed partly by platelet content of the separated plasma of 30 g/l after 500 ml of treatment and 10 g/l after 2,500 ml. Major side effects in the patients were not observed. The new differential filtration system already fulfills the demands of a ready-to-use CE marked rheohemapheresis system but improvements in the details of the treatment protocol are still required and under way. PMID- 11501572 TI - Release of microparticles in LDL apheresis. AB - Particle contamination of blood always takes place in extracorporeal systems and few studies have been conducted to evaluate potential risks. Particle concentration was measured in the efferent blood line on original equipment for two established LDL elimination procedures (DALI) (Fresenius) and Liposorber (Kaneka). Acquired data were compared with standards for infusion solutions from European (EP) and American (USP) Pharmacopoeia. All values were well below the given limits. Even in extreme situations (>20 pump stops) particle concentration did not exceed the standards. Considering an average treated blood volume of 7.31 for the DALI-System and 17.01 for Liposorber (long term clinical studies) the absolute amount of particles infused per treatment was 167,000 (DALI) and 465,000 (Liposorber) particles > or = 2 microm. PMID- 11501574 TI - AB0/O terminology. PMID- 11501573 TI - Leukoreduction of sickle cell trait blood: an unresolved issue. AB - With the increasing demand for leukoreduction of blood components and the implementation of universal leukoreduction in several countries, the problems associated with leukocyte filtration of sickle cell trait blood have been revisited. Currently, there is no unified standard practice for sickle cell trait donors. Different blood centers adopt different policies. While some defer these donors from red cell component donation, some do not. Some screen all ethnic African donors for hemoglobin S (Hb S), others do not. Furthermore, there are differences in views of whether sickle cell trait red cells should be considered as equivalent to non-sickle cell trait red cells. Some blood centers do not give red cells from a sickle cell trait donor to the newborn or patients undergoing general anesthesia as a preventative measure. In this presentation, we discuss the epidemiology of the sickle gene, the sickling process, problems associated with leukoreduction of sickle cell trait whole blood and red cells, and some unresolved issues concerning donor referral and the usage of sickle cell trait blood. PMID- 11501575 TI - Epigenetics and breast cancer. PMID- 11501576 TI - Above and within the genome: epigenetics past and present. AB - Epigenetic regulation involves the maintenance of a particular state of gene expression--most commonly, repression--in the face of repeated mitosis, and frequently meiosis. Remarkably, changes in such heritable expression states occur without an alteration of the primary DNA sequence. We present a brief history of research in epigenetics, beginning with pioneering work in the 1950s by B. McClintock and R. A. Brink on maize kernel color inheritance. We describe the complex biochemistry of DNA methylation--the molecular basis of most epigenetic regulation in mammalian genomes--and review data connecting it to targeted modification and remodeling of chromatin structure. Several prominent examples of epigenetically regulated loci--X chromosome inactivation, imprinting, repetitive DNA silencing, and aberrant methylation patterns in neoplasia--are reviewed along with a description of our current understanding of the underlying molecular mechanisms. A common theme that emerges is the complex integration of epigenetic regulatory pathways with the chromatin infrastructure over target DNA loci. PMID- 11501577 TI - Genes, chromatin, and breast cancer: an epigenetic tale. AB - The production of heritable changes in gene expression is the driving force in the development and progression of breast cancer. Such changes can result from mutations or from epigenetic events such as hypermethylation of DNA and hypoacetylation of histones. Histone acetylation and DNA methylation are major determinants of chromatin structure, and chromatin structure is a primary regulator of gene transcription. Cancer cells frequently contain both mutated genes and genes with altered expression due to one or more epigenetic mechanisms. This review describes the epigenetic changes that disrupt normal chromatin architecture and modify the expression of key genes in breast cancer cells. The structural integrity of the latter genes is usually intact, but their expression has been substantially altered due to methylation in their promoter region or deacetylation of histones that interact with their promoter region or both mechanisms. Genes affected by epigenetic changes in breast cancers include HoxA5, p21WAF, gelsolin, BRCA1, BRCA2, E-cadherin, steroid hormone receptors, and retinoic acid receptor II. Because these epigenetic modifications are usually reversible by treatment with certain drugs, they represent vulnerabilities in the cancer cell that can be exploited as novel targets for new prevention and therapeutic strategies. PMID- 11501578 TI - Role of DNA methylation and histone acetylation in steroid receptor expression in breast cancer. AB - DNA methylation is an epigenetic modification that is associated with transcriptional silencing of gene expression in mammalian cells. Hypermethylation of the promoter CpG islands contributes to the loss of gene function of several tumor related genes, including estrogen receptor a (ER) and progesterone receptor (PR). Gene expression patterns are also heavily influenced by changes in chromatin structure during transcription. Indeed both the predominant mammalian DNA methyltransferase (DNMTI), and the histone deacetylases (HDACs) play crucial roles in maintaining transcriptionally repressive chromatin by forming suppressive complexes at replication foci. These new findings suggest that epigenetic changes might play a crucial role in gene inactivation in breast cancer. Further, inhibition of DNA methylation and histone deacetylation might be a therapeutic strategy in breast cancer, especially for those cancers with ER and PR negative phenotypes. PMID- 11501579 TI - Epigenetic downregulation of the retinoic acid receptor-beta2 gene in breast cancer. AB - A growing body of evidence supports the hypothesis that the retinoic acid receptor beta2 (RAR-beta2) gene is a tumor suppressor gene which induces apoptosis and that the chemopreventive and therapeutic effects of retinoids are due to induction of RAR-beta2. During breast cancer progression, RAR-beta2 is reduced or even lost. It is known from studies of other tumor-suppressor genes that methylation of the 5'-region is the cause of loss of expression. Several groups demonstrated that this is also true for the RAR-beta2 in breast cancer by treating breast cancer cell lines with a demethylating agent and examining expression of the RAR-beta2 gene in response to a challenge with retinoic acid. Studies using sodium bisulfite genomic sequencing as well as methylation specific PCR showed that a number of breast cancer cell lines as well as breast cancer tissue showed signs of methylation. The RAR-beta2 gene was unmethylated in non neoplastic breast tissue as well as in other normal tissues. A combination of retinoic acid with demethylating agents as well as with histone deacetylase inhibitors acts synergistically to inhibit growth. This review presents data that suggest that treatment of cancer patients with demethylating agents followed by retinoic acid may offer a new therapeutic modality. Both the time of commencement of chemoprevention and the choice of substances that are able either to prevent de novo methylation or to reverse methylation-caused gene silencing may be important considerations. PMID- 11501580 TI - The contribution of epigenetic changes to abnormal centrosomes and genomic instability in breast cancer. AB - The centrosome is the major microtubule organizing center of the cell and as such it plays an important role in cytoskeletal organization and in the formation of the bipolar mitotic spindle. Centrosome defects, characterized by abnormal size, number, and microtubule nucleation capacity, are distinguishing features of most high grade breast tumors and have been implicated as a possible cause for the loss of tissue architecture and the origin of mitotic abnormalities seen in solid tumors in general. Centrosome defects arise through uncoupling of centriole duplication and the cell cycle as a result of either genetic alterations or through physical or chemical perturbation of centrosome function. Centrosomes manifest unique epigenetic properties whereby positional or structural information can be propagated through somatic cell lineages by way of nongenetic pathways. Because aberrant centrosome function can result in chromosomal instability, these properties may have important implications for the origin of malignant breast tumors. PMID- 11501581 TI - It takes a tissue to make a tumor: epigenetics, cancer and the microenvironment. AB - How do normal tissues limit the development of cancer? This review discusses the evidence that normal cells effectively restrict malignant behavior, and that such tissue forces must be subjugated to establish a tumor. The action of ionizing radiation will be specifically discussed regarding the disruption of the microenvironment that promotes the transition from preneoplastic to neoplastic growth. Unlike the highly unpredictable nature of genetic mutations, the response of normal cells to radiation damage follows an epigenetic program similar to wound healing and other damage responses. Our hypothesis is that the persistent disruption of the microenvironment in irradiated tissue compromises its ability to suppress carcinogenesis. PMID- 11501582 TI - Epigenetic changes accompanying human mammary epithelial cell immortalization. AB - Acquisition of immortality may be an early and crucial step in malignant progression. We hypothesize that acquisition of unlimited growth potential in individual human mammary epithelial cells (HMEC) requires inactivation of several distinct negative growth constraints as well as reactivation of a mechanism to maintain telomeres on chromosomes. Some of the heritable changes that occur during HMEC immortalization, i.e., loss of expression of cyclin dependent kinase inhibitors p16INK4a and p57KIP2, loss of TGFbeta-mediated growth inhibition, and derepression of telomerase, appear to occur without identifiable mutations in the genes and pathways involved. The absence of mutations, combined with the fact that the changes are often incremental over several cell generations even in clonal populations indicates that some changes associated with immortalization can be epigenetic. We have used the term "conversion" to describe the gradual epigenetic process in chemical carcinogen-immortalized HMEC that leads to activation of telomerase, stabilization of telomere length, and ability to grow uniformly well in the presence or absence of TGFbeta. Characterization of the epigenetic mechanisms involved in immortalization may uncover additional factors that drive tumor progression, and that may be responsive to novel forms of intervention. PMID- 11501584 TI - Dermot Hedley (Derek) Williamson (1929-1998). PMID- 11501583 TI - Loss of chromosomal integrity in human mammary epithelial cells subsequent to escape from senescence. AB - The genomic changes that foster cancer can be either genetic or epigenetic in nature. Early studies focused on genetic changes and how mutational events contribute to changes in gene expression. These point mutations, deletions and amplifications are known to activate oncogenes and inactivate tumor suppressor genes. More recently, multiple epigenetic changes that can have a profound effect on carcinogenesis have been identified. These epigenetic events, such as the methylation of promoter sequences in genes, are under active investigation. In this review we will describe a methylation event that occurs during the propagation of human mammary epithelial cells (HMEC) in culture and detail the accompanying genetic alterations that have been observed. PMID- 11501585 TI - Interleukin-1beta production in periradicular lesions in a human immunodeficiency virus/acquired immune deficiency syndrome model compared with a noninfected host. AB - This study elucidates the role of interleukin-1 (IL-1) in developing periradicular lesions in immunocompetent and immunocompromised (human immunodeficiency virus/acquired immune deficiency syndrome) hosts. Eight cats were immunosuppressed with steroids before infection with feline immunodeficiency virus (FIV). Eight uninoculated cats served as controls. Periradicular lesions were induced around the canine teeth. At 1 and 4 wk periradicular exudate was sampled via the root canals. IL-1beta levels were measured with ELISA. Data were analyzed using the Mann-Whitney U test and the Wilcoxon signed rank test. Statistically significant differences existed in cytokine levels between the FIV and non-FIV groups (p < 0.001). Cytokines were below detectable levels in the FIV group. A significant decrease in IL-1beta levels at 4 wk compared with 1 wk occurred in the non-FIV group (p < 0.05). In conclusion decreased IL-1beta production was obtained in the FIV group. In the non-FIV group decreases in IL 1beta levels were encountered at the chronic stage of the periradicular lesion compared with the acute stage. PMID- 11501586 TI - Qualitative description of a new preparation technique: the balanced-force motion using the Endographe. AB - The forces and torque applied during the balanced-force motion technique of root canal preparation were measured with the Endographe. Five endodontists prepared 100 extracted teeth, all straight and with similar root anatomy. The assignment of "tooth practitioner" was randomized. Gates-Glidden burs were used to prepare the coronal part and ensure apical patency for the other instruments. The balanced-force technique with conventional instruments was then used for the body of the canal. For the apical part either the watch-winding pull with conventional instruments or the reversed balanced-force technique with manual GT files was used. The Endogrammes were similar among endodontists for the coronal phase and showed low vertical forces in contrast with the lateral forces. For the body the Endogrammes were highly specific and described perfectly the different steps of instrument work. For the apical part the Endogrammes of the conventional instruments showed a lack of vertical forces. The Endogrammes of the manual GT files showed that torque was applied inversely to that of the conventional instruments. Force application was similar. In conclusion these objective data indicate the correct way of working with the different techniques incorporated into the balanced-force motion technique. PMID- 11501587 TI - Response of human pulpal tissue to orthodontic extrusive applications. AB - The aim of this study was to investigate histologically the human pulp response to orthodontic extrusive force applications. In group 1, 20 teeth were extruded with the use of a fixed sectional orthodontic appliance. In group 2, 20 teeth were extruded by means of 1/4-inch 4.5-oz elastics. The test periods were 10 and 40 days. None of the teeth in the groups showed any inflammatory reactions or reparative dentin formation at the test periods. Five teeth from the elastics group and two teeth from the sectional appliance group showed large and numerous pulp stones in the serial sections (17.5% of all cases). Seven teeth from the sectional appliance group and two teeth from the elastics group showed odontoblast aspiration into the dentin tubules (22.5%). The results of the study showed that the extrusive forces applied in this study did not cause significant pathological changes in human pulp tissue. PMID- 11501588 TI - Radicular temperatures associated with thermoplasticized gutta-percha. AB - Thermoplasticized gutta-percha has been used to obturate root canals. The continuous wave of condensation technique uses the System B Heat Source with the choice of different-sized pluggers. The purpose of this study was to measure the temperatures within the root canal and on the root surface at different radicular levels while using the System B Heat Source. Fine, Fine-Medium, and Medium pluggers were evaluated at temperature settings of 200 degrees C, 250 degrees C, and 300 degrees C. The Obtura II gutta-percha delivery system following the manufacturer's instructions and ultrasonically thermoplasticized gutta-percha were used for comparative purposes. The highest mean temperature change on the internal root surface was 74.19 degrees C with the system B at the 6 mm level (6 mm coronal to working length) when the Fine-Medium plugger was set at 300 degrees C. The lowest mean temperature change on the internal root surface was 2.09 degrees C at the 0 mm level (at working length) when the F plugger was set at 200 degrees C. With the Obtura II, the lowest mean internal temperature change was 5.22 degrees C at the 0 mm level, whereas the highest mean internal temperature change was 26.63 degrees C at the 6 mm level. With ultrasonic lateral compaction the lowest mean internal temperature change was 5.01 degrees C at the 0 mm level, whereas the highest mean internal temperature change was 28.95 degrees C at the 6 mm level. At no time did the System B, the Obtura II, or ultrasonic delivery of warm gutta-percha exceed an increase of 10 degrees C at any thermocouple level on the external root surface. PMID- 11501589 TI - Influence of structure on nickel-titanium endodontic instruments failure. AB - The purpose of this work was to investigate the process history on fracture life of nickel-titanium endodontics files. The results are based on microstructural investigations of nickel-titanium engine-driven rotary instruments based on X-ray diffraction, scanning electron microscopy, and microhardness tests. Endodontic files are very work-hardened, and there is a high density of defects in the alloy that can disturb the phase transformation. The microhardness Vickers confirmed these observations (dislocations and precipitates). The X-rays show that experimental spectrum lines are extended, typical of a distorted lattice. The surface state of the endodontic files (scanning electron microscopy) is an important factor in failure and fracture initiation. PMID- 11501590 TI - Adhesive sealing of the pulp chamber. AB - The purpose of this in vitro study was to evaluate quantitatively the ability of four different filling materials to seal the orifices of root canals as a secondary seal after root canal therapy. Forty extracted human molar teeth were used. The top of pulp chambers and distal halves of the roots were removed using an Isomet saw. The canal orifices were temporarily sealed with a gutta-percha master cone without sealer. The pulp chambers were then treated with a self etching primer adhesive system (Clearfil SE Bond), a wet bonding system (One Step), a 4-methacryloyloxyethyl trimellitate anhydride adhesive system (C&B Metabond), or a reinforced zinc oxide-eugenol (IRM). The specimens were randomly divided into four groups of 10 each. A fluid filtration method was used for quantitative evaluation of leakage. Measurements of fluid movement were made at 2 min intervals for 8 min. The quality of the seal of each specimen was measured by fluid filtration immediately and after 1 day, 1 wk, and 1 month. Even after 1 month the resins showed an excellent seal. Zinc oxide-eugenol had significantly more leakage when compared with the resin systems (p < 0.05). Adhesive resins should be considered as a secondary seal to prevent intraorifice microleakage. PMID- 11501591 TI - Regional bond strengths of adhesive resins to pulp chamber dentin. AB - Microleakage of oral microorganisms, which can occur due to the lack of sealing ability of permanent restorative materials, may cause failure of root canal treatments. Although a great deal of research has been done on sealing enamel and coronal dentin with resins, little research has been done on the adhesion of resins to the walls of pulp chambers. The purpose of this study was to evaluate regional bond strengths of two adhesive systems to the walls of pulp chambers. A section was made horizontally through the middle of the pulp chamber of extracted human third molars to divide the chamber into upper and lower halves. The pulp tissue was removed and the tooth segments were then divided into treatment subgroups. The pulp chambers were bonded with C&B Metabond (Parkell) or One-Step (Bisco), with or without 5% NaOCI pretreatment. The microtensile bond strengths of these resins to four different pulp chamber regions (bottom, wall, roof, and pulp horn areas) were then measured using an Instron machine. The data were expressed in MPa and were analyzed by a three-way ANOVA. Statistically significant differences were found among the test groups (p < 0.001). One-Step produced higher bond strengths to all pulp chamber regions except the floor, compared with C&B Metabond. The results indicated that high bond strengths can be achieved between adhesive resins and the various regions of the pulp chamber. This should permit the use of a thick layer of unfilled resin along the floor of the pulp chamber and over the canal orifices as a secondary protective seal after finishing root canal therapy. PMID- 11501592 TI - The endodontic cube: a system designed for evaluation of root canal anatomy and canal preparation. AB - An extensively redesigned muffle system is presented incorporating improved design features, such as rigid external fixation and machined internal indexing to enable a more accurate, clinically relevant, and reproducible evaluation of root canal anatomy before and after preparation. Given the ability to directly observe and quantify changes in the root canal system, the information obtained using this design is comprehensive. In each tooth pretreatment evaluation provides the ideal control, reinforcing the suitability of this technique. PMID- 11501593 TI - Philosophies and practices regarding the management of the endodontic smear layer: results from two surveys. AB - The purpose of this investigation was to ascertain the prevailing attitudes within the endodontic community regarding smear layer removal. The first survey asked the philosophies and techniques currently being taught to predoctoral dental students and postdoctoral endodontic residents in the United States, whereas the second surveyed some of the members of the American Association of Endodontists currently practicing in the United States. Findings from these surveys revealed that more than three-fourths of the dental students and nearly two-thirds of the endodontic residents are not being taught routine smear layer removal. Slightly more than one-half of endodontists responded they practice smear layer removal. These surveys indicate that there is no clear consensus in the endodontic community, either academically or clinically, as to whether the smear layer should be removed or be allowed to remain before obturation of the root canal space. PMID- 11501594 TI - Prophylactic treatment of dens evaginatus using mineral trioxide aggregate. AB - Two case reports with dens evaginatus are presented. Each patient had one tooth affected. There was a prominent tubercle on the occlusal surface of the mandibular second premolar. Under local anesthesia and rubber dam isolation a partial pulpotomy was conducted and mineral trioxide aggregate was placed. After 6 months the teeth were removed as part of planned orthodontic treatment. Histological examination of these teeth showed an apparent continuous dentin bridge formation in both teeth, and the pulps were free of inflammation. These cases show that mineral trioxide aggregate can be used as an alternative to existing materials in the proplylactic treatment of dens evaginatus. PMID- 11501595 TI - Conservative management of an accidentally resected root: a 3-year follow-up. AB - Despite all the effort by dentists to provide high levels of dental care in clinical practice, there is always the possibility of accidents occurring. This report describes a referred case that presented with irreversible pulpitis in tooth #19. The situation developed after accidental resectioning of the distal root of tooth #19 during the surgical extraction of tooth #18 by another dentist. The resultant open apex in the distal root of tooth #19 was managed conservatively. A positive apical stop was prepared at the root end of the distal root and sterile calcium hydroxide powder was used to create an apical barrier against which a gutta-percha filling was condensed. The treatment appeared successful at 3-year follow-up. PMID- 11501596 TI - Zebra XXI, Part 2. Cementoblastoma. PMID- 11501597 TI - Statin therapy in the new millennium: is there room for improvement? PMID- 11501598 TI - Newer risk markers and surrogate endpoints in atherosclerosis management. AB - Risk markers that allow improved and individualized assessment of atherosclerotic disease risk and response to treatment are needed. Current candidate markers include cell adhesion molecules, such as intercellular adhesion molecule-1 and E selectin, and inflammatory markers, such as C-reactive protein; advances in genomics and proteomics will suggest additional candidate markers. Noninvasive imaging procedures such as electron-beam computed tomography and magnetic resonance imaging also show considerable promise for monitoring disease status and response to treatment, and ultimately could provide surrogate endpoints for clinical trials of therapeutic interventions. PMID- 11501599 TI - A new statin: a new standard. AB - Numerous studies have demonstrated that treatments designed to reduce low-density lipoprotein cholesterol (LDL-C) can reduce the risk of coronary heart disease (CHD) events in the setting of either primary or secondary prevention. The rationale for aggressive lowering of LDL-C, supported by large observational studies, is the concept that no threshold exists below which reductions fail to provide additional benefit. The statins are widely considered first-line therapy for preventing CHD events because these agents yield the greatest reductions in LDL-C. However, many patients do not achieve target LDL-C levels with the currently available statins. Newer, more effective statins may permit the benefits of aggressive LDL-C reduction to be extended to larger numbers of patients. A novel, highly efficacious statin, rosuvastatin (Crestor, AstraZeneca group of companies), is currently undergoing clinical investigation. Dose-ranging studies in hypercholesterolemic patients have shdwn that rosuvastatin produces significant, dose-dependent decreases in LDL-C when compared with placebo. Reductions have ranged from 34% at a dose of 1 mg/day to 65% at 80 mg/day. This agent has been found to be well tolerated across the range of doses studied. Phase III studies indicate that rosuvastatin is more effective than atorvastatin, pravastatin, and simvastatin in improving the atherogenic lipid profiles of hypercholesterolemic patients, and more effective than atorvastatin in improving the atherogenic lipid profiles of patients with heterozygous familial hypercholesterolemia. Overall, these findings suggest that rosuvastatin is a promising new medication for the treatment of dyslipidemias. PMID- 11501600 TI - Therapies on the horizon for cholesterol reduction. AB - Statins are powerful agents for the reduction of low-density lipoprotein cholesterol (LDL-C) and reduction of cardiovascular risk. Newly developed statins with increased potency, such as rosuvastatin (Crestor) and NK-104 (in earlier clinical development), are capable of achieving marked LDL-C reductions. Cholesterol-lowering agents with mechanisms of action distinct from those of the statins are in active development. These include bile transport inhibitors, such as improved bile acid-absorbing resins and specific inhibitors of the ileal Na+/bile acid cotransporter. There are also specific inhibitors of cholesterol absorption, such as ezetimibe, which may provide cholesterol lowering that is additive to that achieved with statin treatment. Another approach is to reduce cardiovascular risk by modifying atherosclerotic processes within the arterial wall, as represented by the acyl CoA:cholesterol acyltransferase (ACAT) inhibitor avasimibe; ACAT inhibitors may reduce atherosclerotic lesions by inhibiting macrophage cholesterol storage. PMID- 11501601 TI - What do the statin trials tell us? AB - The results of five large-scale, randomized, placebo-controlled trials, involving nearly 31,000 subjects, attest to the benefits of statins in the prevention of coronary events. Several key observations can be made on the basis of the evidence from these investigations. Of primary importance is the fact that statins reduce coronary event rates in patients with or without coronary heart disease. The percentage reduction in risk increases with each successive year of statin therapy. Moreover, the risk reduction is proportional to the reduction in low-density lipoprotein cholesterol. Subgroup analyses have demonstrated that the efficacy of statins extends to specific subgroups of patients, including women, people with diabetes, and older individuals. These agents also reduce the risk of stroke and transient ischemic attacks in patients with coronary disease. Both the randomized trials and widespread clinical experience have confirmed that statins are safe and do not increase the risk of cancer or mortality. Several markers of atherosclerotic risk are ameliorated by statins, although the clinical significance of this observation remains under investigation. The broad range of the therapeutic effects of statins yields safe, effective management of hypercholesterolemia in current practice while also providing a foundation for additional therapeutic refinements in the future. PMID- 11501602 TI - How do we achieve optimal cardiovascular risk reduction? AB - Optimizing coronary heart disease (CHD) risk reduction requires the application of clinical evidence to patient care, as well as the refinement of risk assessment. Clinical evidence indicates that most patients are not treated to optimal low-density lipoprotein (LDL) cholesterol goals. Despite the efficacy of statin therapy in reducing the incidence of CHD, many treated patients still experience CHD events. Targeting other lipid factors such as high-density lipoprotein cholesterol and triglycerides may augment the risk reduction achieved by lowering LDL cholesterol. Refined global risk assessment can lead to more accurate determinations of absolute risk and to the identification both of high risk patients needing aggressive intervention and intermediate-risk patients who appear to be at low risk. Previous global risk assessment measures failed to identify a substantial proportion of primary prevention patients who would benefit from therapy. However, revised guidelines issued by the National Cholesterol Education Program introduce new criteria for more precise risk assessment and advocate use of the Framingham scoring system to calculate absolute risk. Although intensified treatment is recommended for high-risk patients, cost considerations may limit drug therapy for some lower-risk individuals. PMID- 11501603 TI - What happened to the Wolff-Parkinson-White syndrome? PMID- 11501604 TI - Spinal cord stimulation for chronic intractable angina pectoris: a unified theory on its mechanism. AB - The use of spinal cord stimulation (SCS) for chronic intractable anginal pain was first described in 1987. Numerous studies have demonstrated its efficacy in improving exercise tolerance, decreasing frequency of anginal episodes, and prolonging time to electrocardiographic signs of ischemia. This review will examine the potential mechanisms of this antianginal effect and propose a unified hypothesis explaining it. The effect of SCS involves a mutual interaction of decreased pain, decreased sympathetic tone, and a likely redistribution of myocardial blood flow to ischemic regions. Spinal cord stimulation reduces the transmission of nociceptive impulse via the spinothalamic tract due to an enhanced release of gamma aminobutyric acid (GABA) from dorsal horn interneurons. Improvement of myocardial blood flow at the microvascular level has been demonstrated by positron emission tomography (PET). A decreased sympathetic tone has been shown by norepinephrine kinetics, tests of sympathetic reflexes, and the use of ganglionic blockers. We hypothesize that SCS exerts its beneficial effects by decreasing pain and decreasing sympathetic tone, the result of which is decreased myocardial oxygen consumption along with an improved myocardial microcirculatory blood flow. PMID- 11501605 TI - Obesity is associated with premature occurrence of acute myocardial infarction. AB - BACKGROUND: The American Heart Association has classified obesity as a major modifiable risk factor for coronary artery disease, but its relationship with age at presentation with acute myocardial infarction (AMI) is poorly documented. HYPOTHESIS: The study was undertaken to evaluate the impact of obesity on age at presentation, and on in-hospital morbidity and mortality in patients with AMI. METHODS: Our analysis includes a consecutive series of 906 Olmsted County patients (mean age 67.7 years, 51% male) admitted with AMI to the Mayo Clinic Coronary Care Unit (CCU). The patients were entered into the Mayo CCU Database, a prospective registry of data pertaining to patients admitted to the Mayo Clinic CCU with AMI. Age at AMI occurrence and in-hospital morbidity and mortality were noted. RESULTS: Obese patients (body mass index [BMI] >30) with AMI were significantly younger than patients with AMI in the overweight (BMI 25-30) and normal-weight (BMI < 30) groups (62.3+/-13.1 vs. 66.9+/-13.2 and 72.9+/-13.4, respectively. p < 0.001). Obesity and overweight status were associated with male gender, diabetes mellitus, hypercholesterolemia, and smoking history; however, after multivariate adjustment for these risk factors, excess weight and premature AMI remained significantly associated. Compared with normal-weight patients, overweight patients presenting with AMI were 3.6 years younger (p < 0.001, confidence interval [CI] 1.9-5.4) and obese patients 8.2 years younger (p < 0.001, Cl 6.2-10.1). No significant increase in in-hospital morbidity and mortality was seen. CONCLUSION: In this population-based study, overweight and obese status are independently associated with the premature occurrence of AMI, but not with an increased incidence of in-hospital complications. PMID- 11501606 TI - Exceptional prevalence of interatrial block in a general hospital population. AB - BACKGROUND: Interatrial block (IAB: P wave > or = 110 ms) is a strong correlate of left atrial (LA) enlargement and an important predictor of supraventricular tachyarrhythmias, notably atrial fibrillation and flutter. It is surprising that, despite its association with arrhythmias and its effects on the electromechanical properties of the left atrium, there is widespread neglect of this common abnormality. HYPOTHESIS: The study was undertaken to investigate the prevalence of IAB in a general hospital population. METHODS: We prospectively evaluated the electrocardiograms of 1,000 consecutive adult patients. analyzed for P-wave duration. RESULTS: Our results showed a very high prevalence of IAB (41.1% of patients in sinus rhythm and 32.8% of all patients). As expected, it was more common in patients aged > 60 years. CONCLUSIONS: Given this unusually high prevalence of IAB in hospital patients and its ominous portents (LA enlargement. thrombosis and embolism, arrhythmias), physicians should be aware of its frequency and computer software should be programmed to recognize it. PMID- 11501607 TI - Long-term follow-up after deferring angioplasty in asymptomatic patients with moderate noncritical in-stent restenosis. AB - BACKGROUND: Many patients with in-stent restenosis (ISR) are angina-free, but the optimal treatment for these patients remains uncertain. HYPOTHESIS: In cases with asymptomatic moderate noncritical ISR. deferral of the intervention may be safe and associated with favorable clinical outcome. METHODS: We evaluated the long term clinical outcome of asymptomatic patients (Group 1, n = 98) with moderate noncritical ISR (< 70% diameter stenosis) after intervention was deferred, and compared it with that of patients (Group 2, n = 655) without restenosis. After repeat angioplasty was deferred, all patients were treated medically and later underwent angioplasty only in the case of clinical recurrence. RESULTS: Baseline characteristics were similar between the two groups. Clinical follow-up was available in all patients at 26.3+/-15.9 months. Twenty patients died during the follow-up: 1 in Group 1 and 19 in Group 2. Target lesion revascularization was performed in 3 patients in Group 1 and 11 patients in Group 2 during follow-up (p = NS), and new lesion revascularization in 2 patients in Group 1 and 27 patients in Group 2 (p = NS). Event-free survival rate (cardiac death, nonfatal myocardial infarction, repeat revascularization) was 86.7+/-6.1% in Group 1 and 84.8+/-2.2% in Group 2 at the end of follow-up (p = NS). Major adverse cardiac events were only associated with the presence of diabetic mellitus (hazards ratio 2.65, 95% confidence interval [CI] 1.48-4.73, p<0.01). The percentage of patients receiving antianginal medication was similar between the two groups at the end of the study (p = NS). CONCLUSIONS: Asymptomatic patients with moderate noncritical ISR have a good prognosis and similar clinical outcome as those without ISR, suggesting that it may be safe to defer repeat angioplasty in these patients until angina recurrence. PMID- 11501608 TI - Development and validation of the ITG Health-Related Quality-of-Life Short-Form measure for use in patients with coronary artery disease. Integrated Therapeutics Group. AB - BACKGROUND: Available coronary artery disease (CAD)-specific health-related quality-of-life (HRQL) measures are not ideally suited for routine clinical practice. We report development of a valid and reliable CAD-specific short-form measure. HYPOTHESIS: The Integrated Therapeutics Group (ITG) CAD-specific short form HRQL measure is reliable and valid for assessing the health status of patients with CAD. METHODS: In all, 409 patients (mean age 62.4 years) completed the 24-item CAD-specific HRQL questionnaire. Factor analysis was used to identify the scaling of the 24 CAD-specific items. Analysis of variance was used to reduce the number of items within each scale, and reliability (Cronbach's alpha), discriminant validity, and ceiling and floor effects of short-form scales were evaluated. Short-form scales were compared with the longer scales using relative validity coefficients. The CAD severity was assessed using New York Heart Association criteria, physician assessment, electrocardiogram results, the number of myocardial infarctions, and the number of CAD-specific medications. Clinical validity of short-form scales was then assessed based on their ability to discriminate across severity levels of these clinical criteria. RESULTS: Four scales were identified: Extent of Chest Pain, Functioning and Well-Being, Activities Level-Physical, and Activities Level-Social. The 24-item questionnaire was reduced to a 13-item short form, with reliability exceeding 0.70 for all four scales. Relative validity estimates comparing short-form to original scales ranged from 0.68 to 2.58. Mean scores varied significantly (p < 0.05) by clinical severity, supporting the discriminant validity of the ITG CAD short-form scales. CONCLUSIONS: The ITG CAD short form (used alone or with a general HRQL measure) is valid and practical for assessing patients with CAD. PMID- 11501609 TI - Event-free survival in patients after an acute coronary event with exercise induced normalization of the T-wave. AB - BACKGROUND: Risk stratification of patients with unstable angina or non-Q-wave myocardial infarction (MI) is an unresolved clinical problem. The prognostic value of T-wave normalization (TWN) during exercise has not been studied in this group of patients. HYPOTHESIS: Event-free survival in clinically stable patients after an acute coronary event without ST-segment elevation can be predicted by the presence of exercise-induced TWN. METHODS: Sixty-five patients (43 men and 22 women, mean age 62+/-10 years) entered the study. The diagnosis of unstable angina and non-Q-wave MI was made in 40 and 25 patients, respectively. A treadmill exercise test was performed in all patients after clinical stabilization. The patients were divided into three groups: those with negative baseline T waves and exercise-induced TWN (Group 1); those with negative baseline T waves, but without TWN (Group 2); and those with positive baseline T waves (Group 3). The patients were followed up for 6 months. RESULTS: During follow-up, serious cardiovascular complications occurred in 15 (23%) patients. These included exacerbation of ischemic heart disease (14 patients) and acute MI (1 patient). Event-free survival was greater in patients in Group 1 (95%) than in those in Group 2 (68%, p < 0.034) or Group 3 (71%, NS). Among all patients studied, exercise-induced TWN was predictive of event-free survival with a sensitivity of 38% and a specificity of 93%. CONCLUSIONS: In clinically stable patients after an acute coronary event without ST-segment elevation, exercise induced TWN is a specific but n ot sensitive predictor of event-free survival after 6 months. PMID- 11501610 TI - A mitral valve myxoma prolapsing into the left ventricular outflow tract. PMID- 11501611 TI - George Ralph Mines: victim of self-experimentation? PMID- 11501612 TI - Female lower urinary tract symptoms: pharmacotherapeutic consequences. PMID- 11501613 TI - Novel role for alpha1-adrenergic receptor subtypes in lower urinary tract symptoms. PMID- 11501614 TI - Alpha-blockers and lower urinary tract function: more than smooth muscle relaxation? PMID- 11501615 TI - Clinical pharmacology of alpha1A selective and nonselective alpha1-blockers. PMID- 11501616 TI - New roles for muscarinic receptors in the pathophysiology of lower urinary tract symptoms. PMID- 11501617 TI - Modern pharmacotherapy of urge urinary incontinence in the USA: tolterodine and oxybutynin. PMID- 11501618 TI - Issues of uroselectivity in male and female LUTS. Summary of the meeting. PMID- 11501619 TI - Tissue remodeling of rat pulmonary artery in hypoxic breathing. I. Changes of morphology, zero-stress state, and gene expression. AB - The remodeling of the pulmonary arterial tissue in response to a step change of the oxygen concentration in the gas in which a rat lives was recorded as function of time and function of O2 concentration. Three steps of changing from 20.9% to 17.2%, 13.6%, and 10% O2 were imposed. Earlier work in our laboratory has shown that pulmonary arterial tissue remodeling is significant in the first 24 h after a step change of oxygen tension. Hence we made measurements in this period. Furthermore, data were obtained for tissue remodeling of circumferential and axial lengths of the pulmonary arteries. We recorded the activities of gene expressions in the lung tissues by microarray, determined the dose response curves of gene expression in the homogenized whole lungs with respect to four levels of O2 concentration, and obtained the time courses of gene expression in the lung parenchyma in 30 days after a step decrease of O2 concentration from 20.9% to 10%. We would like to suggest that the correlation of gene expression with physiological function parameters, i.e., time, O2 tension, blood pressure, opening angle, wall thicknesses, etc., is the way to narrow down the search for specific genes for specific physiological functions. PMID- 11501620 TI - Tissue remodeling of rat pulmonary artery in hypoxic breathing. II. Course of change of mechanical properties. AB - When cells and the matrix of a tissue remodel, the mechanical properties of the tissue do change. The mechanical properties are expressed by constitutive equations. In this article the remodeling of the constitutive equation of the pulmonary artery is studied. The remodeling was induced in a rat breathing a gas whose oxygen concentration was suddenly decreased as a step function of time and maintained constant (17.2%, 13.6%, or 10%) afterwards. Since the mathematical form of the constitutive equation has been identified in earlier papers, we need to determine only the elastic constants that change in the process of tissue remodeling. We consider arteries subjected to blood pressure and longitudinal stretch, and limit ourselves to two-dimensional problems involving only circumferential and longitudinal stress and strain. In the neighborhood of an in vivo state, the perturbations of stresses and strains are related by linear, anisotropic, tensor equations involving three elastic constants: the incremental Young's modulus in the circumferential direction Ythetaz, that in the longitudinal direction Yzz, and the cross modulus Ythetaz. Over a 24 h period, changes of Ythetatheta between 164 and 187 kN/m2, Yzz between 64 and 92 kN/m2, and Ythetaz between 61 and 88 kN/m2 are statistically insignificant. PMID- 11501621 TI - Role of tissue hypoxia in cerebrovascular regulation: a mathematical modeling study. AB - This paper presents a mathematical model of cerebrovascular regulation, in which emphasis is given to the role of tissue hypoxia on cerebral blood flow (CBF). In the model. three different mechanisms are assumed to work on smooth muscle tension at the level of large and small pial arteries: CO2 reactivity, tissue hypoxia, and a third mechanism necessary to provide good reproduction of autoregulation to cerebral perfusion pressure (CPP) changes. Using a single set of parameters for the mechanism gains, assigned via a best fitting procedure, the model is able to reproduce the pattern of pial artery caliber and CBF under a large variety of physiological stimuli, either acting separately (hypoxia, CPP changes, CO2 pressure changes) or combination (hypercapnia+hypoxia; hypercapnia+hypotension). Furthermore, the model can explain the increase in CBF and the vasoconstriction of small pial arteries observed experimentally during hemodilution, ascribing it to the decrease in blood viscosity and to the antagonistic action of the flow-dependent mechanism (responsible for vasoconstriction) and of hypoxia (responsible for vasodilation). Finally, the interaction between hypoxia and intracranial pressure (ICP) has been analyzed. This interaction turns out quite complex, leading to different ICP time patterns depending on the status of the cerebrospinal fluid outflow pathways and of intracranial compliance. PMID- 11501622 TI - Role of the mechanical properties of tracheobronchial airways in determining the respiratory resistance time course. AB - A physiologically based simulation model of breathing mechanics was considered in an attempt to interpret and explain the time course of input respiratory resistance during the breathing cycle, observed in recent studies on ventilated patients. The model assumes a flow-dependent Rohrer resistance for the upper extrathoracic airways and volume-dependent resistance and elastance for the intermediate airways. A volume-dependent resistance describes the dissipative pressure loss in the lower airways, and two constant elastances represent lung and chest wall elasticity. Simulated mouth flow and pressure signals obtained in a variety of well-controlled conditions were used to analyze total respiratory resistance and elastance estimated by an on-line algorithm based on a time varying parameter model. These estimates were compared with those provided by classical estimation algorithms based on time-invariant models with two, three, and four parameters. The results show that the four-parameter model is difficult to identify, while the three-parameter one offers no substantial advantage for estimating input resistance with respect to the more simple two-parameter model. In contrast, the time-varying approach provides good on-line estimates of the simulated end-expiration and end-inspiration resistances. These values provide further information of potential clinical utility, with respect to time-invariant models. For example, the results show that the difference between the end expiration and end-inspiration resistance increases when obstructions shift from the upper to the lower airways. The similarity of the results obtained with measured and simulated data indicates that, in spite of its simplicity, the simulation model describes important physiological mechanisms underlying changes in respiratory input resistance, specifically the mechanical properties of intermediate airways. PMID- 11501623 TI - A noninvasive estimation of hypernasality using a linear predictive model. AB - The pronunciation of a speaker with a defective soft palate is marked by hypernasality and an operation may be necessary to repair the defective soft palate to reduce this hypernasality. An assessment of hypernasality is necessary to quantify the effect of the surgery. The current clinical methods for assessing hypernasality are uncomfortable or require expensive equipment. In this paper, a new quantitative method is proposed to estimate hypernasality. This method requires only a microphone and a personal computer equipped with a sound card. Zeros in the frequency response of the vocal tract system are one of the major characteristics of hypernasality. The proposed method made use of the fact that a linear predictive model with a typical order for the human vocal tract system is not accurate when the vocal tract system has zeros in its frequency response. Hypernasality was estimated by comparing the distance between the sequences of linear predictive cepstrum of low- and high-order linear predictive models. The proposed method provides a better correlation (0.58) with nasalance measured by a nasometer than Teager method (0.44) for all the data. Furthermore, the proposed method showed higher correlation of 0.84 than 0.71 of the Teager method for data with a nasalance higher than 35%. Since the proposed method needs only digitized speech data, it is much less invasive and provides an easy and cost-effective evaluation of hypernasality. PMID- 11501624 TI - Minimally invasive estimation of systemic vascular parameters. AB - A cardiovascular parameter estimator to identify the systemic vascular parameters was developed using an extended Kalman filter (EKF) algorithm. Measurements from a ventricular assist device (VAD) and arterial pressure were used in the estimator. The systemic vascular parameters are important indices of heart condition. However, obtaining these parameters usually requires invasive measurements, which are difficult to obtain under most clinical environments. Including a VAD model into the estimator and using the signals from a VAD to identify the cardiovascular parameters for VAD patients would minimize the need for indwelling sensors. This paper illustrates the use of a Novacor left ventricular assist system (LVAS) model with a cardiovascular model in the estimator to identify the systemic vascular parameters: characteristic resistance, blood inertance at the aorta, systemic compliance, and systemic resistance. Performance of the estimator was evaluated using data from a computer simulation and from a mock circulatory system experiment. Robustness of the estimator to the available measurements was also described. The estimation results showed that the estimates converged with reasonable accuracy in a limited time when the LVAS pump volume and arterial pressure were used as measurements. These parameter estimates can provide additional diagnostic information for patient and device monitoring and can be used for future VAD control development. PMID- 11501625 TI - Deterministic chaos and noise in three in vitro hippocampal models of epilepsy. AB - Recent reports have suggested that chaos control techniques may be useful for electrically manipulating epileptiform bursting behavior in neuronal ensembles. Because the dynamics of spontaneous in vitro bursting had not been well determined previously, analysis of this behavior in the rat hippocampus was performed. Epileptiform bursting was induced in transverse rat hippocampal slices using three experimental methods. Slices were bathed in artificial cerebrospinal fluid containing: (1) elevated potassium ([K+]o= 10.5 mM), (2) zero magnesium, or (3) the GABAA-receptor antagonists bicuculline (20 microM) and picrotoxin (250 microM). The existence of chaos and determinism was assessed using two different analytical techniques: unstable periodic orbit (UPO) analysis and a new technique for estimating Lyapunov exponents. Significance of these results was assessed by comparing the calculations for each experiment with corresponding randomized surrogate data. UPOs of multiple periods were highly prevalent in experiments from all three epilepsy models: 73% of all experiments contained at least one statistically significant period-1 or period-2 orbit. However, the expansion rate analysis did not provide any evidence of determinism in the data. This suggests that the system may be globally stochastic but contains local pockets of determinism. Thus, manipulation of bursting behavior using chaos control algorithms may yet hold promise for reverting or preventing epileptic seizures. PMID- 11501626 TI - Identification of time-varying Hammerstein systems from ensemble data. AB - In this paper, we describe a new technique to identify rapidly time-varying Hammerstein systems from ensembles of input-output realizations. The technique involves two steps. A correlation approach is first used to obtain initial estimates of the linear subsystem parameters for every sampling time. An iterative optimization algorithm is then employed to produce final estimates of the system parameters. The input does not need to be white. The technique was tested on simulated data and was found to produce excellent results under realistic conditions. PMID- 11501627 TI - Editorial: transferring the helm. PMID- 11501628 TI - 3D-VIEWER: an atlas-based system for individual and statistical investigations of the human brain. AB - 3D-VIEWER is a new software tool for neurosurgical planning and population studies. It is based on digitized three-dimensional brain atlases derived from standard stereotactic atlases that can be adapted to an individual's brain and shown as a series of displayed images. If the patient's brain has been imaged in different modalities, the standardized anatomical information can be adapted to the individual images, which will bring the images into registration. The 3D VIEWER can be used as a tool for combining multimodal information from the same patient. In addition, several tools are available that allow oblique views of anatomical structures or the view along the intended trajectory during a neurosurgical intervention. Furthermore, using the atlas transformation matrices, anatomical information can be determined when comparing an individual's brain to the anatomy of the atlas brain. Thus, standardized anatomical information from the atlas can be introduced into individual images. This standardization is used to perform individual-group and group-by-group comparisons between patients and normal controls in anatomical studies. PMID- 11501629 TI - A four-dimensional approach to medical image retrieval. AB - Image retrieval is faced with different problems. Few proposed solutions consider the heterogeneity of user competence (e.g. physicians, researchers, students, etc.) and the necessity of a high expressive power for medical image description. This paper presents a solution designed for medical imaging retrieval. It consists of providing a four-dimensional approach able to integrate different views of the medical image, to deal with complex medical queries. Via use of the Spatial Knowledge Model, we propose hybrid (textual and visual) and customizable interfaces, appropriate for non-computer scientist users. Our prototype, accessible through the Web, has been tested by a group of medical students and physicians and gives very satisfactory results. PMID- 11501630 TI - Paradigms for recording cognitive brain electrical activity. AB - Human event-related potentials reflect cognitive processing, and are normally elicited by external events, such as acoustic sounds or visual stimuli. As such they provide an opportunity to study normal and abnormal brain function noninvasively, at sub-second resolution. Advances in multimedia technology permit specialists in informatics and neuropsychology to co-operate in the design and implementation of paradigms, which influence ERP components. The paper illustrates the progression from standard paradigms such as the auditory oddball, which can be used to study memory update through to contingent negative variation and three condition, visual paradigms which can be used to study cognitive and emotional responses. Data from a study investigating the comparative processing of target pictures and words illustrate how external stimuli influence the later cognitive potentials. PMID- 11501632 TI - Probabilistic record linkage: relationships between file sizes, identifiers and match weights. AB - This study investigates relationships between file sizes, amounts of information contained in commonly used record linkage variables, and the amount of information needed for a successful probabilistic linkage project. We present an equation predicting the amount of information needed for a successful linkage project. Match weights for variables commonly used in record linkage are measured using artificially created databases. Linkage algorithms were successful when the sum of minimum weights for variables used in a linkage exceeded the predicted cutoff. Linkage results were acceptable when this sum was near the predicted cutoff. This technique enables researchers to determine if enough information exists to perform a successful probabilistic linkage. PMID- 11501631 TI - Influence of the method of data collection on the documentation of blood-pressure readings with an Anesthesia Information Management System (AIMS). AB - The influence of methods for record keeping on the documentation of vital signs was assessed for the Anesthesia Information Management System (AIMS) NarkoData. We compared manually entered blood-pressure readings with automatically collected data. These data were stored in a data-base and subsequently evaluated and analyzed. The data sets were split into two groups, "manual" and "automatic". We evaluated the effect of automatic data collection on the incidence of corrected data, data validity and data variation. Blood-pressure readings of 37,726 data sets were analyzed. We could assess that the method of documentation did influence the data quality. It could not be assessed whether the incorrectness of data during automatic data gathering was caused by artefacts or by the anesthesiologist. PMID- 11501633 TI - Partitioning an object-oriented terminology schema. AB - Controlled medical terminologies are increasingly becoming strategic components of various healthcare enterprises. However, the typical medical terminology can be difficult to exploit due to its extensive size and high density. The schema of a medical terminology offered by an object-oriented representation is a valuable tool in providing an abstract view of the terminology, enhancing comprehensibility and making it more usable. However, schemas themselves can be large and unwieldy. We present a methodology for partitioning a medical terminology schema into manageably sized fragments that promote increased comprehension. Our methodology has a refinement process for the subclass hierarchy of the terminology schema. The methodology is carried out by a medical domain expert in conjunction with a computer. The expert is guided by a set of three modeling rules, which guarantee that the resulting partitioned schema consists of a forest of trees. This makes it easier to understand and consequently use the medical terminology. The application of our methodology to the schema of the Medical Entities Dictionary (MED) is presented. PMID- 11501634 TI - A prospective evaluation of the medical consultation system CADIAG-II/RHEUMA in a rheumatological outpatient clinic. AB - To evaluate the performance of CADIAG-II/RHEUMA as consultant in the primary evaluation of patients visiting a rheumatological outpatient clinic, a CADIAG II/RHEUMA consultation was done for 54 patients and the list of generated diagnostic hypotheses was compared to each clinical discharge diagnosis. For 26 of a total of 126 rheumatological discharge diagnoses, no matching CADIAG II/RHEUMA diagnosis was available. 94% of all other discharge diagnoses were found in the list of CADIAG-II/RHEUMA hypotheses, 82% among the first third of the list of hypotheses and 48% among the first five hypotheses. We identified the following factors limiting the ability of CADIAG-II/RHEUMA to generate a comprehensive and correctly ranked list of diagnostic hypotheses: (1) a large percentage of patients with early stages of not clearly identified rheumatological conditions; (2) the limited number of CADIAG-II/RHEUMA diagnoses compared to the large number of known rheumatological conditions; (3) the fact that rheumatological diseases are rarely characterized by a single pathognomonic feature but are usually diagnosed by combinations of rather unspecific findings. PMID- 11501635 TI - Attitudes towards information system security among physicians in Croatia. AB - To examine attitudes about information system security among Croatian physicians a cross-sectional study was performed on a representative sample of 800 Croatian physicians. An anonymous questionnaire comprising 21 questions was distributed and statistical analysis was performed using a chi-square test. A 76.2% response rate was obtained. The majority of respondents (85.8%) believe that information system security is a new area in their work. In general, physicians are not informed about European directives, conventions, recommendations, etc. Only a small number of physicians use personal computers at work (29%). Those physicians who have a personal computer use it mainly for administrative reasons. Most healthcare institutions (89%) do not have a security manual and the area of information system security is left to individual interest and initiative. Only 25% of physicians who have a personal computer use any type of password. A high percentage of physicians (22%) has never thought about the problem of personal data being used by organizations (e.g. police, banks) without legal background; a small, but still significant percentage of physicians (5.6%) has even agreed with such use. Results indicate that for the vast majority of physicians, information system security is a new area in their daily work, one which is left to individual interest and initiative. They are not familiar with the ethical, technical and legal backgrounds which have been defined for that area within the Council of Europe and the European Union. New aspects: This is the first study performed in Central and Eastern Europe dealing with information system security, performed on a representative nationwide sample of all the physicians. PMID- 11501636 TI - Migration of legacy mumps applications to relational database servers. AB - An extended implementation of the Mumps language is described that facilitates vendor neutral migration of legacy Mumps applications to SQL-based relational database servers. Implemented as a compiler, this system translates Mumps programs to operating system independent, standard C code for subsequent compilation to fully stand-alone, binary executables. Added built-in functions and support modules extend the native hierarchical Mumps database with access to industry standard, networked, relational database management servers (RDBMS) thus freeing Mumps applications from dependence upon vendor specific, proprietary, unstandardized database models. Unlike Mumps systems that have added captive, proprietary RDMBS access, the programs generated by this development environment can be used with any RDBMS system that supports common network access protocols. Additional features include a built-in web server interface and the ability to interoperate directly with programs and functions written in other languages. PMID- 11501637 TI - Advantages of long observation in episode-oriented electronic patient records in family practice. AB - From 1985-2000, 58 Dutch family physicians (FPs) of the Transition Project collected ICPC-coded data on 47, 2451 episodes of care, first in paper records for direct encounters only, later with a complete electronic patient record (EPR) for all (direct and indirect) encounters. Based on these data, the effects of a long observation period and the inclusion of all encounters (both direct and indirect) in the EPR were studied. Long observation periods in EPRs appear to have important advantages for patient documentation and the assessment of the content of family practice. Comparison of data from a one-year versus a four-year observation period showed a shift in utilization rates. In a long observation period, visiting patients appeared to make less demand on care, while the content of the FP's care for selected chronic diseases was reflected more realistically. The inclusion of all indirect encounters in an EPR (as compared to the previous inclusion of direct encounters only) resulted in more than twice the number of coded entries per listed patient, and thus led to a major shift in perspective on the FP's involvement in patient care. PMID- 11501638 TI - Putting the pieces together: using "off-the-shelf" software to safely transfer medical data. AB - We describe a new computer-based, network-driven system for administering, transferring, and evaluating reports of spontaneous adverse drug reactions and serious adverse events. The system consists of three regional centers, one coordinating center, and a central database at the national drug regulatory authority. In view of its high level of data security, reliability, easy expandability, and inexpensiveness due to the use of standard shareware products, the system can serve as a model for a clinical health information network. PMID- 11501639 TI - Nonparametric frontier model as a tool for exploratory analysis of hospital stays. AB - Diagnosis-related groups (DRG) were introduced in 1995 to the Belgian hospital financing system. Trimming rules are generally used when mean length of stay (LOS) is estimated by DRG. This paper proposes the use of frontier models instead of trimming rules. These models allow to take into account the characteristics of the patients, to rank hospital stays, and to indicate stays presenting discrepancy between the patient's characteristics and the resources consumed. The analysis is done with the nonparametric Free Disposal Hull (FDH) model and the method developed by Wilson to detect extreme observations, when defining the frontier is adapted to analyze large databases. PMID- 11501640 TI - More up-to-date monitoring of long-term survival rates by cancer registries: an empirical example. AB - A few years ago, a new method of survival analysis, denoted period analysis, has been developed to derive more up-to-date estimates of long-term survival rates of cancer patients, but this method has rarely been applied so far. Using empirical examples from the Saarland Cancer Registry, we illustrate how seriously traditional long-term survival estimates may lag behind survival expectations of newly diagnosed cancer patients in the case of recent improvement in prognosis, and to what extent this problem may be reduced by period analysis of survival. We conclude that period analysis should be more widely used for deriving more up-to date long-term survival estimates. PMID- 11501641 TI - Retrieval-oriented design of clinical research forms. AB - Computerized clinical forms are subject to a wide variety of different requirements. They have to allow detailed documentation and must be user friendly. State-of-the-art applications for design permit clinicians themselves to create their own forms as needed, with the various variables presented in different ways depending on their intended use. Often, however, only aspects of clinical documentation are considered, with no thought being given to subsequent data retrieval. This article presents guidelines for the retrieval-oriented design of clinical forms. It discusses where anticipatory measures for structuring forms are easier to accomplish than complex data linkage at the time of retrieval and analysis. PMID- 11501642 TI - Application of resampling techniques to the statistical analysis of the Brier score. AB - We investigated the application of resampling techniques to the statistical analysis of the Brier score (B), and extended them to the statistical comparison of two Bs derived from the same set of patients. The resampling techniques are helpful in the statistical analysis of B, and there are almost no differences between the jackknife method and the bootstrap method in this analysis. Thus, we believe that B should be used more often as an index to evaluate probabilistic judgments in the case in which the data sets for the assessment are "degenerate" as the "receiver operating characteristic data sets." PMID- 11501643 TI - A measurement model of disease severity in absence of a gold standard. AB - In the absence of a gold standard, we propose the use of confirmatory factor analysis for the quantification of agreement between diagnostic measurements and the true disease severity. The essential assumption is conditional independence of diagnostic measurements adjusted for the severity of the disease. However, depending on the number of measurements available, the method works even if some of them are conditionally dependent. We illustrate the method using an example related to glaucoma eye disease. PMID- 11501644 TI - Water sorption-desorption test and moisture accumulation test for functional assessment of atopic skin in children. AB - Sorption-desorption and moisture accumulation tests are simple and quick methods for the in vivo functional analysis of stratum corneum hydration kinetics. The aim of this study was to evaluate the hydration dynamics of the uninvolved and affected skin of children with atopic dermatitis and to compare them with the skin of healthy children. The study investigated 45 children. The dynamic tests were performed using the corneometer CM820. Numerical parameters were calculated. With the sorption-desorption test, eczematous skin showed lower water accumulation during the sorption phase, whereas water was released more slowly during the desorption phase. With the moisture accumulation test, increases in water accumulation velocity and in water accumulation were observed in atopic children. Dynamic tests showed that the stratum corneum of unaffected atopic skin was less hydrated but more easily hydratable than normal skin. Conversely, despite a lower absorption capability, eczematous skin showed a greater avidity to retain water. New functional parameters (water-sorption capacity and accumulated water decay) are proposed to describe more precisely the hydration kinetics of eczematous skin. PMID- 11501645 TI - Effect of moisturizers on skin susceptibility to irritants. AB - Moisturizers are used for the treatment of dry and irritated skin. The benefit of moisturizers when used on normal skin has recently been challenged, since an earlier study indicated that the increased hydration that follows long-term use of moisturizers on normal skin may facilitate penetration of irritants. The aim of the present study was to evaluate short-term use of 2 different moisturizers used on normal skin: cream A (high lipid content) and B (moderate/low lipid content). Nineteen healthy volunteers applied the moisturizers on the upper arm/forearm 3 times daily for 5 days, while the other upper arm/forearm served as symmetrical control. The day after moisturizer treatment was stopped the skin was challenged with a patch test of sodium lauryl sulphate. Skin reactions were evaluated by bioengineering measuring methods and clinical scoring. Skin response to sodium lauryl sulphate was increased on moisturizer-treated arms compared to controls for one of the moisturizer (cream A), while this was not statistically significant for the other moisturizer (cream B). Data confirm previous indications that some moisturizers when used on normal skin may increase skin susceptibility to irritants. PMID- 11501646 TI - A questionnaire for the assessment of pruritus: validation in uremic patients. AB - A questionnaire was constructed for the evaluation and measurement of pruritus. The questionnaire, based on the short form of the McGill Pain Questionnaire, was tested in 145 patients suffering from uremic pruritus and currently undergoing hemodialysis treatment in 3 centers. The newly developed questionnaire proved to be reliable and provided valid data on the sensory, affective and overall intensity of uremic pruritus. The data suggest that uremic pruritus tends to be prolonged, frequently intense and a major source of distress to the patient. Dialysis was not found to influence the pruritus. The questionnaire may also be useful in pruritus secondary to other causes. PMID- 11501647 TI - Relationship between season and diagnoses of melanocytic tumours. AB - The frequency of various diagnoses of pigmented lesions, their distribution on various body sites and the seasonal variations in their diagnoses were studied according to the specimen of pigmented lesions sent to a laboratory for histopathological examination. From the total number of 18,402 investigated lesions, 7,682 were excised from males and 10,720 from females. Melanoma was diagnosed in 2.5% of all excised tumours more often during the summer months. Lighter clothing and greater awareness during summer may explain this finding. Additional factors such as a sun-induced melanocytic activation in naevi cannot be ruled out. Dysplastic naevi, which represented 26% of all naevi in this study, were excised less often during summer. Presuming that the majority of the north German population acquires the highest ultraviolet (UV) doses of the year during summer, it seems unlikely that the histological features of dysplasia in naevi are related to short-term UV activation. The localization of dysplastic naevi did not correspond to the body-site distribution of melanoma according to sex. This finding does not indicate the dysplastic naevus as a possible precursor lesion of malignant melanoma. PMID- 11501648 TI - Treatment of psoriasis in the Nordic countries: a questionnaire survey from 5739 members of the psoriasis associations data from the Nordic Quality of Life Study. AB - The data from a questionnaire-based study of 5,739 members of the psoriasis associations of Denmark, Finland, Iceland, Norway, Sweden and the Faeroe Islands showed that the two most commonly used active agents were topical steroids (89.7% total use and 49.4% present use) and calcipotriol (73.1% total use and 35.8% present use), with only small variations between the countries. Marked differences between the countries were, however, found within all other types of psoriasis therapy, including the so-called alternative treatments. Significant priorities varied between the different countries. The use of dithranol in Finland was almost twice the average. While 14.2% of Danish members had received grenz-rays within the last week only 0.1% of the Finns had been given the same treatment. Psoralen plus ultraviolet A (PUVA) was being used by 13.1% of the Finnish psoriatics compared with 3.8% of Danes, while PUVA was almost non existent on the Faeroe Islands. The use of non-PUVA phototherapy was highest in Norway and Sweden. Almost 10% of the Danes were presently on methotrexate, which was used far more than etretinate/acitretin or cyclosporine. In contrast, Finnish patients more often received etretinate than other systemic agents, and in Iceland there was a higher present use of cyclosporine than of etretinate. The popularity of alternative therapies was highest in Iceland, where 26.6% had taken such medication during the last week. The results of the study suggest that different treatment patterns should be taken into consideration when discussing the prognosis of psoriasis in different countries. PMID- 11501649 TI - Patch testing of nickel sulfate and potassium dichromate with a standardized ready-to-use test system gives highly reproducible results: a double-blind multicentre study. AB - There is still some doubt about the reproducibility of patch tests. A sound assessment needs optimized and unbiased studies. This study analysed the results of a double-blind multicentre study with nickel sulfate and potassium dichromate patch tests attached synchronously to both sides of the back of patients with a history of nickel allergy, conducted with a highly standardized randomized test system (TRUE-test). Out of 589 patients tested, a total of 388 had responded with allergic reactions to nickel sulfate and 130 to potassium dichromate. The reproducibility of positive nickel (dichromate) patch tests was 99.2% (90.8%). The reaction index was also calculated, which relates the number of allergic reactions obtained with a test preparation to the number of questionable and irritant reactions; the reaction index can range from -1 (questionable and irritant reactions only occur) to 1 (allergic reactions only occur). For nickel sulfate the reaction index was 0.91, but it was only 0.23 for potassium dichromate, as a result of considerably more questionable reactions. In conclusion, a highly synchronous reproducibility of results can be achieved by using a well-standardized patch-test system, especially with nickel sulfate. However, distinct allergens and test systems need to be evaluated separately. PMID- 11501651 TI - Extensive human papillomavirus type 7-associated orofacial warts in an immunocompetent patient. AB - Human papillomavirus (HPV) type 7 is frequently found in butchers' warts and has been demonstrated in oral and facial warts of HIV-infected patients. The reservoirs of HPV7 and the route of transmission are still unclear. Here we describe an HIV-negative, otherwise healthy patient with extensive, recurrent orofacial papillomatosis whose immune status proved to be normal and who had no history of meat handling. HPV7 L1 gene DNA that differed in 3 point mutations from the HPV7 prototype could be detected in 2 morphologically distinct, perioral lesions by different PCR protocols. In situ hybridization confirmed the presence of HPV7 DNA in the nuclei of vacuolated cells of the granular layer. Our data show that HPV7 can lead to perioral, spiky warts and brownish plaques in immunocompetent patients who had never been working as a meat or fish handler. PMID- 11501650 TI - HPV-related cancer susceptibility and p53 codon 72 polymorphism. AB - Conflicting results regarding the association of a polymorphism at codon 72 of the p53 tumour suppressor gene and susceptibility to develop human papilloma virus (HPV)-associated cervical cancer have been published over the last year, implicating differences in ethnic background, sample origin, sample size and/or detection assay. The material for this study was collected in the identical geographical region as for 2 previous reports with contradictory results regarding the association of codon 72 genotype with squamous cell cancer (SCC). We have used an alternative detection assay, based on pyrosequencing technology, that interrogates the variable position by the accuracy of DNA polymerase. In addition to cervical clinical specimens from SCC, HPV16- and HPV18-infected adenocarcinoma cases as well as cervical intraepithelial neoplasia (CIN) were investigated. No significant association was found between p53 codon 72 genotype and the risk to develop adenocarcinoma, SCC or CIN in the Swedish population. PMID- 11501652 TI - Pyodermatitis-pyostomatitis vegetans: a clinical course of two decades with response to cyclosporine and low-dose prednisolone. AB - Pyodermatitis-pyostomatitis vegetans is a rare, polymorphous inflammatory disorder of the skin and oral mucosa first described by Hallopeau in 1898. On the skin papules, pustules and reddish brown annular vegetating plaques develop, most frequently in the intertriginous areas. In the mouth, yellowish flat ulcerations arise, typically in the shape of "snail tracks". The association with inflammatory bowel disease is very common. An unusual case with a chronic relapsing course of 2 decades is presented. Gastrointestinal inflammation was absent. Prednisolone in high and medium doses suppressed most lesions. Various attempts with other drugs (dapsone, isotretinoin, azathioprine) to reduce the corticosteroid dose failed. This is the first report of the successful treatment of pyodermatitis-pyostomatitis vegetans with cyclosporin A, which proved to be highly effective in this regard. The unknown aetiopathology of pyodermatitis pyostomatitis vegetans is discussed. PMID- 11501653 TI - A case of chondrodermatitis nodularis chronica helicis with an autoantibody to denatured type II collagen. PMID- 11501654 TI - Epithelioid sarcoma: a frequently misdiagnosed neoplasm. PMID- 11501655 TI - Infantile acropustulosis successfully controlled with topical corticosteroids under damp tubular retention bandages. PMID- 11501656 TI - Autoinvolutive photoexacerbated tinea corporis mimicking a subacute cutaneous lupus erythematosus. PMID- 11501657 TI - Pharyngitis and exanthema caused by Arcanobacterium haemolyticum. PMID- 11501658 TI - Atopic dermatitis: increased prevalence and the influence of birth, siblings and maternal factors. PMID- 11501659 TI - Molluscum contagiosum on a tattoo. PMID- 11501660 TI - Dowling-Degos disease affecting the vulva. PMID- 11501661 TI - Pemphigus vulgaris in two sisters. PMID- 11501662 TI - Multiple paraneoplasia in the absence of a detectable tumour. PMID- 11501663 TI - Local reactions associated with subcutaneous injections of both beta-interferon 1a and 1b. PMID- 11501664 TI - Uselessness of topical calcipotriol as monotherapy for acrodermatitis continua of Hallopeau. PMID- 11501665 TI - Recurrence of fungal nail disease and the dissociation of relapse from re infection. PMID- 11501666 TI - Epidermal p53 response and repair of thymine dimers in human skin after a single dose of ultraviolet radiation: effects of photoprotection. AB - A cellular p53 response, DNA repair enzymes and melanin pigmentation are important strategies utilized by skin keratinocytes against impairment caused by ultraviolet radiation (UVR). In this study a double-immunofluorescence technique was used to investigate UVR-induced thymine dimers and p53 protein simultaneously. Four healthy volunteers were irradiated on both sides of their buttock skin with a single dose of solar-simulating UVR. One side was pretreated with a topical sunscreen. Biopsies from different time-points were immunostained for visualization of thymine dimers, p53 and proliferation. One single physiological dose of UVR generated widespread formation of thymine dimers throughout the epidermis 4h after irradiation. The level of thymine dimers decreased over time and was followed by a p53 response in the same cells. A late proliferative response was also found. The formation of thymine dimers, the p53 response and the late proliferative response were partially blocked by topical sunscreen. Large inter-individual differences in the kinetics of thymine dimer formation and repair as well as in the p53 response were evident in both sunscreen-protected and unprotected skin. PMID- 11501667 TI - Urocanic acid isomers do not modulate intracellular calcium and cyclic AMP in human natural killer cells. AB - Ultraviolet irradiation influences natural killer cell function both in vitro and in vivo. The postulated ultraviolet photoreceptor in the epidermis, urocanic acid, has been reported to depress the cytotoxic activity of human natural killer cells. Therefore, this study investigated whether this would occur through specific second messengers, using a radioimmunoassay for intracellular adenosine 3',5'-cyclic monophosphate (cAMP) and Fluo-3 staining plus flow cytometry for free calcium. Both isolated lymphocytes and enriched CD16+ cells were used. A combination of the trans- and cis-isomers of urocanic acid (200 microg/ml) induced cAMP in both CD16+ and CD16- cells, but individual, stereospecific effects were not demonstrable. Urocanic acid did not induce significant changes in calcium levels in lymphocytes, or natural killer cells alone or conjugated to K562 target cells. Evidently, the biochemistry of urocanic acid-mediated natural killer-cell modulation is complex, and the cellular receptor(s) and specific signal transduction pathway(s) mediating the biological effects of urocanic acid remain elusive. PMID- 11501668 TI - Role of Th2 cytokines, RANTES and eotaxin in AIDS-associated eosinophilic folliculitis. AB - The pathogenesis of AIDS-associated eosinophilic folliculitis is still unknown. The expression of chemokines and Th2-type cytokines is increased in other conditions associated with tissue eosinophilia and in allergic reactions. We evaluated the mRNA expression by reverse transcriptase polymerase chain reaction of two Th2 cytokines (interleukin-4 and interleukin-5) and of two chemokines (RANTES and eotaxin) in the skin of 6 patients with AIDS-associated eosinophilic folliculitis; the tissue localization of eotaxin was shown by immunohistochemistry. We demonstrated the increased expression of interleukin-4, interleukin-5, RANTES and eotaxin in lesional skin of the patients compared to normal skin of HIV+ individuals. We concluded that a Th2 pattern is present in AIDS-associated eosinophilic folliculitis. The cytokine milieu in this disease may favour a Th2 immune response to an unknown antigen, whereby RANTES and eotaxin act in synergy with interleukin-4 and interleukin-5 to mediate tissue inflammation. PMID- 11501669 TI - Neutrophil-related host response in hidradenitis suppurativa: a pilot study in patients with inactive disease. AB - Host-defence defects in hidradenitis suppurativa patients have been suspected, but not proven. Activated neutrophils can destroy the surrounding tissues by a release of reactive oxygen species and active proteases. Peripheral neutrophils from 15 female patients (mean age 46, range 27-57 years) in an inactive state of their hidradenitis suppurativa, were studied and compared with 15 age-matched healthy female controls. There were no significant differences between patients and controls in the assessments of intracellular elastase activity, total content of antigenic elastase or release of elastase. Furthermore, no differences were found in total content and membrane expression of the receptors measured. The generation of free oxygen radicals, after stimulation with the protein kinase C activator phorbol myristate acetate, was significantly higher in the patients than in the controls, while there was no difference after Fc-receptor-mediated stimulation. Dysfunctional neutrophils might be involved in the pathogenesis of hidradenitis suppurativa, but the findings should be interpreted with caution because of the small number of observed cases. PMID- 11501670 TI - The Tol-Pal proteins of the Escherichia coli cell envelope: an energized system required for outer membrane integrity? AB - The outer membrane of gram-negative bacteria acts as a barrier against harmful lipophilic compounds and larger molecules unable to diffuse freely through the porins. However, outer membrane proteins together with the Tol-Pal and TonB systems have been exploited for the entry of macromolecules such as bacteriocins and phage DNA through the Escherichia coli cell envelope. The TonB system is involved in the active transport of iron siderophores and vitamin B12, while no more precise physiological role of the Tol-Pal system has yet been defined than its requirement for cell envelope integrity. These two systems, containing an energized inner membrane protein interacting with outer membrane proteins, share similarities. PMID- 11501671 TI - Prion model in yeast. AB - Two unusual phenotypes in Saccharomyces cerevisiae, [PSI+] and [URE3], have been suggested to be due to prion proteins. Various research groups have shown that this is indeed the case and have characterized these yeast prions in more detail. The factors involved in prion formation, such as chaperone protein, and the intramolecular determinants of prion formation have been investigated. The ability of these yeast proteins to form prion is due to modular domains conserved throughout evolution. PMID- 11501673 TI - Genetic characterization of the nonconventional yeast Hansenula anomala. AB - We describe genetic, molecular and taxonomic characteristics of the yeast Hansenula anomala. Pulsed-field gel electrophoresis of chromosomal DNAs from 19 H. anomala strains and related species indicated that H. anomala had a clearly different karyotype. Chromosome length polymorphism of the H. anomala strains was independent of their geographic origin and source of isolation. The strains were classified into four groups of similar karyotypes and one strain showed a unique profile. The sizes of chromosomes ranged from 850 to 3500 kb in different strains. The haploid chromosome number of H. anomala is at least nine. We have found RAPD primers discriminating at both the species and strain levels. All the primers tested except the M13 core sequence generated unique patterns with most strains. The results indicate the usefulness of PCR analysis with primer M13 for identification of the H. anomala species. Screening of the CBS (Utrecht) collection strains of H. anomala showed that they are rather difficult objects for genetic hybridization analysis. The strains have low fertility, viz. very poor sporulation, low mating type activities and, as a rule, nonviable ascospores. The majority of the hybrids obtained are polyploid, probably tetraploid, as judged by the segregation of control auxotrophic markers. Nevertheless, some monosporic cultures of the strains studied, including the biocontrol yeast J121, formed diploid hybrids with regular meiotic segregation of control auxotrophic markers. As a rule, H. anomala isolates are homothallic, showing delayed self-diploidization. Rare stable heterothallic strains of H. anomala also occur. PMID- 11501672 TI - Expression of horizontally transferred gene clusters: activation by promoter generating mutations. AB - The occurrence of promoter-generating mutations allowing the transcription of heterologous genes has been studied in a system based on the plasmid-mediated conjugal transfer of histidine biosynthetic genes from a donor bacterium (Azospirillum brasilense) into a heterologous Escherichia coli mutant population lacking histidine biosynthetic ability and initially unable to recognize the transcriptional signal of the introgressed gene(s). Under selective stressful conditions, His+ revertants accumulated in the E. coli His- culture. The number of His+ colonies was dependent on the time of incubation under selective conditions, the strength of selective pressure, and on the crowding of cells plated; moreover, it was independent of the physiological status of the cell (i.e. the growth phase). Sequence analysis of plasmid DNA extracted from E. coli His+ revertants revealed that single base substitutions in the region upstream of the A. brasilense his operon resulted in an adjustment of the pre-existing sequence that was rendered similar to the E. coli -10 promoter sequence and transcriptable by the host RNA-polymerase. One particular transition (C --> T) was predominant in the His+ revertants. Data presented here indicated that the barriers to the expression of horizontally transferred heterologous genes or operons may be overcome in a short time scale and at high frequency, and supported the selfish operon model on the origin and evolution of gene clusters. PMID- 11501674 TI - Recombinant Listeria strains producing the nontoxic L-chain of botulinum neurotoxin A in a soluble form. AB - Fragments of Clostridium botulinum neurotoxin A (BoNT/A) gene (botA) were expressed in Listeria monocytogenes ATCC10527 to produce the L-chain of the toxin in a soluble form. A shuttle vector pAT19 (EmR) was used to make plasmid pAT-RL containing a botA gene fragment placed under C. botulinum ntnH-gene promoter control. The plasmid also contained a C. botulinum botR/A gene, a positive transcriptional regulator of botA. The cytoplasmic fraction of the L. monocytogenes (pAT-RL) strain was found to contain up to 3 mg/L of a soluble protein of expected size and immunologically positive towards BoNT antibodies. This is the first evidence of heterologous botA gene expression producing a soluble safe derivative of botulinum neurotoxin A needed as a molecular tool for exploratory research in neurosciences as well as a basis for raising protective immunity in humans. PMID- 11501675 TI - Activity against Mycobacterium tuberculosis with concomitant induction of cellular immune responses by a tetraaza-macrocycle with acetate pendant arms. AB - The novel tetraaza-macrocyclic compound 3,7,11-tris(carboxymethyl)-3,7,11,17 tetraaza-bicyclo[11.3.1]heptadeca-1(17),13,15-triene, abbreviated as ac3py14, was investigated for its activity against Mycobacterium tuberculosis and for induction of protective cellular immune responses. Perspective results show that ac3py14 and its Fe3+ 1:1 complex, [Fe(ac3py14)], inhibited radiometric growth of several strains of M. tuberculosis. Inhibition with 25 microg/mL varied from 99% for H37Rv to 80% and above for multiple drug-resistant clinical isolates. The capacity of ac3py14 to elicit a beneficial immune response without cellular apoptosis was assessed and compared to the effects of virulent M. tuberculosis. The present study produces evidence that after stimulation with ac3py14 there was significant production of interferon gamma (IFN-gamma), whereas the production of interleukin-5 (IL-5) remained low, and there was development of a memory population (CD45RO). The level of binding of Annexin V, a marker of apoptosis, was not sufficient to result in toxic effects toward alphabeta and gammadelta T cells and CD14+ macrophages. This preliminary study is the first report of a compound that simultaneously exerts an inhibitory effect against M. tuberculosis and induces factors associated with protective immune responses. PMID- 11501676 TI - Direct detection of thermotolerant campylobacters in chicken products by PCR and in situ hybridization. AB - We have evaluated the use of PCR and fluorescent in situ hybridization (FISH) techniques for the detection of thermotolerant campylobacters in naturally contaminated chicken products. 16S rRNA sequence data was used to design two specific primers and an oligonucleotide probe for PCR and FISH analyses, respectively. The PCR protocol amplified a 439-bp fragment corresponding to a portion of specific 16S RNA gene from thermotolerant campylobacters. The detection range of the PCR assay varied between 10 cells (after enrichment) to 10(2) cells per mL (without enrichment). FISH probes were able to identify thermotolerant Campylobacter species in 'spiked' and 'unspiked' naturally contaminated samples. PCR and FISH were performed on naturally contaminated samples and compared with the isolation of cells on selective media. The in situ hybridization technique was less sensitive than PCR, although its sensitivity of detection was increased considerably after 22 h of enrichment. These results confirm the usefulness of 16S rRNA-based techniques for the direct detection of campylobacters in food samples. PMID- 11501677 TI - Polychlorinated biphenyl degradation activities and hybridization analyses of fifteen aerobic strains isolated from a PCB-contaminated site. AB - Fifteen bacterial strains using biphenyl as sole carbon and energy source, obtained from different positions and depths of a polychlorinated biphenyl (PCB) contaminated area, were analyzed for their basic metabolic phenotypes and subjected to genomic DNA hybridization screening for the presence of well characterized bph operons such as those of Pseudomonas pseudoalcaligenes KF707 and Rhodococcus globerulus P6. Most of the isolates belonged to the gamma subdivision (Pseudomonas stutzeri, P. plutida, P. fluorescens and Vibrio logei species) and to the beta-subdivision (genera Alcaligenes, Comamonas, Ralstonia) of the Proteobacteria. All the isolates were able to cometabolize different low chlorinated PCB congeners. Among the dichlorinated biphenyls tested, a lower degradation capacity was observed for the di-ortho substituted congeners, whereas high levels of degradation were observed for the di-meta and di-para isomers, whether they were chlorinated on one or on both rings. The PCB congeners nonsubstituted in the 2,3 or 2,3 and 3,4 positions were also degraded by most of the isolated strains, which were, however, unable to significantly metabolize PCBs with more than 3 chlorine atoms. Five of the isolated strains were also able to degrade some of the tri- and tetrachlorobiphenyls tested. Southern hybridization analysis showed a strong homology between four of the fifteen isolated strains and the bph operon obtained from P. pseudoalcaligenes strain KF707. Conversely, none of the isolates here examined showed homology with the bph operon of R. globerulus strain P6. In line with this, the KF707 bph probe strongly hybridized with DNA of a significant number of bacterial colonies obtained from selected locations in the contaminated area using biphenyl supplemented minimal medium agar plates. PMID- 11501678 TI - Hydrogen peroxide effects on Streptococcus thermophilus CNRZ368 cell viability. AB - Streptococcus thermophilus CNRZ368 is an anaerobic aerotolerant bacteria and its ability to survive under aerobic growth conditions raises the question of the existence of a putative defence system against oxidative stress. Thus, survival of CNRZ368 in the presence of increasing concentrations of hydrogen peroxide was studied. Moreover, the influence of the physiologic state of the cells, as well as that of a preexposition with sublethal doses of hydrogen peroxide, upon S. thermophilus CNRZ368 survival were determined. The results suggest that S. thermophilus displays a defence system against oxidative stress and that this system is inducible. PMID- 11501679 TI - More on fluvoxamine and enuresis. PMID- 11501680 TI - Haloperidol decanoate in children. PMID- 11501681 TI - Valproic acid and risperidone. PMID- 11501682 TI - Mirtazapine for excessive masturbation in an adolescent with autism. PMID- 11501683 TI - Conversion and dissociative disorders in the Sultanate of Oman. PMID- 11501684 TI - Autistic psychopathy. PMID- 11501685 TI - National Institute of Mental Health research roundtable on prepubertal bipolar disorder. AB - OBJECTIVE: A research roundtable meeting was convened at the National Institute of Mental Health on April 27, 2000, to discuss the existing controversial areas in the diagnosis of bipolar disorder in prepubertal children. METHOD: Invited clinicians and researchers with expertise on bipolar disorder in children were asked to share and discuss their perspectives on diagnostic issues for bipolar disorder in prepubertal children. RESULTS: The group reached agreement that diagnosis of bipolar disorder in prepubertal children is possible with currently available psychiatric assessment instruments. In addition to phenotypes that fit DSM-IV criteria for bipolar I and bipolar II, participants agreed on the existence of other phenotypic possibilities that do not meet diagnostic criteria. Bipolar not otherwise specified (NOS) was recommended as a "working diagnosis" for the non-DSM-IV phenotype. CONCLUSIONS: Bipolar disorder exists and can be diagnosed in prepubertal children. In children who present with both the DSM-lV and non-DSM-IV phenotypes (i.e., those given a diagnosis of bipolar-NOS), assessment should include careful evaluation of all behaviors that are impairing. Moreover, these children should be monitored systematically to explore stability and change over time in diagnosis and impairment. PMID- 11501686 TI - Adaptation of preventive interventions for a low-income, culturally diverse community. AB - OBJECTIVE: To describe essential elements in the adaptation of a prevention approach with a high-risk urban sample, chosen to contrast sharply with the primarily middle-class sample in which it had been originally tested. METHOD: Key elements of a preventive intervention for families with parental depression were adapted for use in the new context. RESULTS: A sequence of alliance-building events was implemented, involving engagement at three levels: community, caregivers, and family. The prevention approach was modified to include an expanded approach to defining depression and resilience; greater flexibility on the part of the clinician; more intensive engagement between clinician and family, with a focus on immediate daily concerns; as well as awareness of cultural issues and responsiveness to the subject's experience of violence. CONCLUSION: Core principles of helping family members to discuss the effects of depression and adversity on family life were affirmed. PMID- 11501687 TI - Olanzapine versus haloperidol in children with autistic disorder: an open pilot study. AB - OBJECTIVES: Conventional neuroleptics ameliorate symptoms in children with autistic disorder; however, they are known to cause dyskinesias. Atypical neuroleptics, including olanzapine, may have less risk for dyskinesia, but their efficacy in autistic disorder is not established. This study was designed to investigate the safety and effectiveness of open-label olanzapine as a treatment for children with autistic disorder by using haloperidol as a standard comparator treatment. METHOD: In a parallel groups design, 12 children with DSM-IV autistic disorder (mean age 7.8+/-2.1 years) were randomized to 6 weeks of open treatment with olanzapine or haloperidol. Mean final dosages were 7.9+/-2.5 mg/day for olanzapine and 1.4+/-0.7 mg/day for haloperidol. Outcome measures included the Clinical Global Impressions (CGI) and the Children's Psychiatric Rating Scale (CPRS). RESULTS: Both groups had symptom reduction. Five of six in the olanzapine group and three of six in the haloperidol group were rated as responders according to the CGI Improvement item. Subjects showed improvement on the CPRS Autism Factor (F1,9 = 24.4, p = .0008). Side effects included drowsiness and weight gain. CONCLUSIONS: The findings suggest that olanzapine is a promising treatment for children with autistic disorder. Further placebo-controlled and long-term studies of olanzapine in autistic disorder are required. PMID- 11501688 TI - Long-term experience with citalopram in the treatment of adolescent OCD. AB - OBJECTIVE: The primary purpose of the study was to describe tolerability and effectiveness of citalopram in the treatment of adolescent obsessive-compulsive disorder (OCD). METHOD: Thirty nondepressed patients (15 females, 15 males) with a mean age of 13.7 years (range 13-18 years) were treated for their OCD with citalopram in an open-label, flexible-dose study (range of dose 20-70 mg; mean dose 46.5 mg). All patients were referred to Aarhus University Hospital. The patients were monitored for 1 to 2 years. The mean total score on the Yale-Brown Obsessive Compulsive Scale (child or adult version) was 28.7 at base-line, 23.3 after 10 weeks of treatment, 20.0 after 6 months, 18.4 after 1 year, and 17.9 after 2 years (from baseline to 2 years of treatment: t = 11.65; p < .001). RESULTS: Seventy percent showed a decrease in total Yale-Brown Obsessive Compulsive Scale score in excess of 35% from baseline to 1 year of treatment. Twenty percent still had a score of greater than 20 after 1 year of treatment, indicating that clinically they still had OCD. Side effects were similar to those reported from the use of other selective serotonin reuptake inhibitors (SSRIs). No patient was excluded because of serious side effects during the 1 year of observation. CONCLUSIONS: The clinical effectiveness and tolerability of citalopram in the long-term treatment seem to be comparable with the observations of other SSRIs in childhood and adolescent OCD. A further, statistically significant reduction is provided by an extended treatment period of up to 1 year. PMID- 11501689 TI - Case study: caudate glutamatergic changes with paroxetine persist after medication discontinuation in pediatric OCD. AB - Proton magnetic resonance spectroscopy (1H-MRS) was used to examine glutamatergic (Glx) abnormalities in the caudate nucleus in pediatric obsessive-compulsive disorder (OCD), associated with severity of illness and response to acute (12 weeks) treatment with paroxetine. In this report, OCD symptoms improved markedly in an 8-year-old girl treated for 14 months with the selective serotonin reuptake inhibitor paroxetine (titrated from 10 to 40 mg/day). Paroxetine dose was then decreased in 10-mg decrements and discontinued without symptom recurrence. Serial 1H-MRS examinations were acquired before and after 12 weeks of paroxetine treatment (40 mg/day) and 3 months after medication discontinuation. A striking decrease in caudate Glx was observed after 12 weeks of treatment which persisted after medication discontinuation. These data provide further support for a reversible glutamatergically mediated dysfunction of the caudate nucleus in OCD that may serve as a pathophysiological and treatment response marker. PMID- 11501690 TI - Measures of psychopathology in children with complex partial seizures and primary generalized epilepsy with absence. AB - OBJECTIVE: This investigation examined psychopathology in 48 children with complex partial seizures (CPS), 39 children with primary generalized epilepsy with absence (PGE), and 59 nonepileptic children, aged 5 to 16 years, by comparing the Child Behavior Checklist (CBCL) and the Schedule for Affective Disorders and Schizophrenia for School-Age Children (K-SADS). METHOD: The CBCL was completed by parents and the K-SADS was administered to both parent and child. RESULTS: The CBCL identified psychopathology in 26% and the K-SADS in 51% of the CPS and PGE patients (kappa = 0.32). The CPS and PGE groups had significantly higher mean CBCL scores, as well as higher rates of psychiatric diagnoses and symptoms of psychopathology, compared with the nonepileptic group. However, the CPS and PGE groups did not differ in these measures. Within each patient group, Full Scale IQ, but not seizure control, was associated with these measures of psychopathology. CONCLUSION: These findings suggest that the K-SADS identifies more children with psychopathology than the CBCL in children with CPS and PGE. PMID- 11501691 TI - Relationship between acute morphine and the course of PTSD in children with burns. AB - OBJECTIVE: To investigate the relationship between the dose of morphine administered during a child's hospitalization for an acute burn and the course of posttraumatic stress disorder (PTSD) symptoms over the 6-month period following discharge from the hospital. METHOD: Twenty-four children admitted to the hospital for an acute burn were assessed twice with the Child PTSD Reaction Index: while in the hospital and 6 months after discharge. The Colored Analogue Pain Scale was also administered during the hospitalization. All patients received morphine while in the hospital. The mean dose of morphine (mg/kg/day) was calculated for each subject through chart review. RESULTS: The Pearson product moment correlation revealed a significant association between the dose of morphine received while in the hospital and a 6-month reduction in PTSD symptoms. Children receiving higher doses of morphine had a greater reduction in PTSD symptoms over 6 months. CONCLUSIONS: This study suggests the possibility that acute treatment with morphine can secondarily prevent PTSD. This result is discussed in terms of the possible effect of morphine on fear conditioning and the consolidation of traumatic memory. PMID- 11501692 TI - Moderators and mediators of long-term adherence to stimulant treatment in children with ADHD. AB - OBJECTIVE: To identify moderators and mediators of long-term adherence to stimulant medication in children with attention-deficit hyperactivity disorder (ADHD). METHOD: Seventy-one children with ADHD were prescribed methylphenidate, followed prospectively on an annual basis for 3 years, and evaluated for adherence to stimulant treatment. The study occurred in Toronto between 1993 and 1997. Adherents were those who took methylphenidate, or another psychostimulant, for 5 or more days per week throughout the follow-up period, except for "drug holidays." Children who adhered at consecutive evaluations were compared with those who did not. Severity of ADHD, presence of oppositional defiant disorder/conduct disorder, learning difficulties, anxiety, age, family dysfunction, and socioeconomic adversity at baseline were investigated as moderators of adherence. Response to treatment at school, measured at 12 months, was investigated as a mediator of adherence. RESULTS: Fifty-two percent of children adhered to stimulant treatment for 3 years. Absence of teacher-rated oppositional defiant disorder, more teacher-rated ADHD symptoms, and younger age at baseline predicted adherence. CONCLUSIONS: Adherence to stimulant medications is a significant factor in the long-term treatment of children with ADHD. PMID- 11501693 TI - Screening for anxiety and depression in early adolescence. AB - OBJECTIVE: To examine the level of diagnostic and discriminative accuracy of three dimensional rating scales for detecting anxiety and depressive disorders in a school-based survey of 9th grade youths. METHOD: Classroom screening instruments, the Center for Epidemiologic Studies-Depression Scale (CES-D), the Revised Children's Manifest Anxiety Scale (RCMAS), and the Multidimensional Anxiety Scale for Children (MASC) were administered to 632 youths from three sites in 1998. On the basis of rating scale results, samples of high-scoring and non-high-scoring youths were invited to participate in a diagnostic interview conducted within 2 months of the screening sessions. RESULTS: MASC scores were most strongly associated with individual anxiety disorders, particularly among females, whereas the CES-D composite score was associated with a diagnosis of major depression, after controlling for comorbid disorders. The RCMAS was least successful in discriminating anxiety and depression. When receiver operator characteristic curves were examined, diagnostic accuracy was moderate. CONCLUSIONS: The ability of the MASC and CES-D to discriminate within and between categorically defined diagnostic groups has important implications for the accurate identification of youths in need of services. PMID- 11501694 TI - Test-retest reliability of anxiety symptoms and diagnoses with the Anxiety Disorders Interview Schedule for DSM-IV: child and parent versions. AB - OBJECTIVE: To examine the test-retest reliability of the DSM-IV anxiety symptoms and disorders in children with the Anxiety Disorders Interview Schedule for DSM IV: Child and Parent Versions (ADIS for DSM-IV:C/P). METHOD: Sixty-two children (aged 7-16 years) and their parents underwent two administrations of the ADIS for DSM-IV:C/P with a test-retest interval of 7 to 14 days. RESULTS AND CONCLUSIONS: Results revealed that the ADIS for DSM-IV:C/P is a reliable instrument for deriving DSM-IV anxiety disorder symptoms and diagnoses in children. The ADIS for DSM-IV:C/P was found to have excellent reliability in symptom scale scores for separation anxiety disorder, social phobia, specific phobia, and generalized anxiety disorder and good to excellent reliability for deriving combined diagnoses of these disorders, as well as using child-only and parent-only interview information. Reliability coefficients were generally similar and, in most instances, superior to those found in previous ADIS-C/P reliability studies. Limitations and directions for future research are discussed. PMID- 11501695 TI - A parent-child relationship scale derived from the child and adolescent psychiatric assessment (CAPA). AB - OBJECTIVE: To examine a measure of children's perception of their relationships with parents. METHOD: The Child and Adolescent Psychiatric Assessment (CAPA) was administered to 114 inpatients (aged 9-18 years) at a tertiary asthma center from 1991 to 1994. Ten items from the CAPA were developed as a separate scale, the Parent-Child Relationship Scale (PCRS). Some subjects (82) were given family and child assessments. RESULTS: The PCRS had good internal reliability (Cronbach alpha = .72). Construct validity was demonstrated by correlations with child and parent report on the Family Assessment Device (r= 0.46 and 0.35; p < .001) and high expressed emotion of the parent (t= 2.89; p < .01). Divergent validity may be evidenced by the fact that the PCRS was not significantly related to high emotional over-involvement. Predictive validity was shown by significant correlations with the total problem scores of parents' (r = 0.28; p < .01) and children's (r = 0.41; p < .001) Achenbach reports, and prediction of CAPA psychiatric diagnosis (OR = 5.83; 95% CI 1.80-22.63). CONCLUSION: The PCRS can potentially be used to assess the child's perspective of the parent-child relationship for research or clinical purposes and deserves further study. PMID- 11501696 TI - Multiple complex developmental disorder: the "multiple and complex" evolution of the "childhood borderline syndrome" construct. AB - OBJECTIVES: To provide an overview of the history, evolution, and nosology of the diagnostic constructs for "borderline syndrome of childhood," also known as "multiple complex developmental disorder." METHOD: The authors synthesized information found via electronic searches of databases (MEDLINE, PsycINFO, Current Contents, Humanities Abstracts, and Social Sciences Abstracts) and bibliographic directed searches. RESULTS: Although early publications (prior to 1980) were either highly anecdotal or lacking in scientific rigor, they were nonetheless noted for their historic value and influence on research trends. The recent publications (1990s) were characterized by more rigorous methodology and greater generalizability. Current classifications, proposals for diagnostic criteria, epidemiological data, and nosological suggestions were summarized. CONCLUSION: The literature supports the creation of a new diagnostic label to describe a population of children whose symptoms are currently subsumed under the labels "borderline" or "multiple complex developmental disorder." A full characterization of the syndrome, including its evolution, would require prospective studies and may differ from the known evolution for personality disorders and/or pervasive developmental disorders. The authors propose a process by which a new nomenclature is derived. PMID- 11501697 TI - Thought disorder in attention-deficit hyperactivity disorder. AB - OBJECTIVE: This study compared thought disorder and associated cognitive variables in attention-deficit hyperactivity disorder (ADHD) and schizophrenia. METHOD: Speech samples of 115 ADHD, 88 schizophrenic, and 190 normal children, aged 8 to 15 years, were coded for thought disorder. A structured psychiatric interview, the WISC-R, the Continuous Performance Test, and the Span of Apprehension task were administered to each child. RESULTS: The ADHD and schizophrenic groups had thought disorder compared with the normal children. However, the subjects with ADHD had a narrower range of less severe thought disorder than did the schizophrenic subjects. The younger children with ADHD and schizophrenia had significantly more thought disorder than did the older children with these diagnoses. IQ, attention, and working memory were associated with thought disorder in the ADHD but not the schizophrenic group. CONCLUSIONS: Thought disorder in childhood is not specific to schizophrenia and reflects impaired development of children's communication skills. PMID- 11501698 TI - Data collection techniques. AB - We have provided an overview of techniques used to assess variables in the applied behavioral sciences. Most of the methods are used by both quantitative/positivist and qualitative/constructivist researchers but to different extents. Qualitative researchers prefer more open-ended, less structured data collection techniques than do quantitative researchers. Direct observation of participants is common in experimental and qualitative research; it is less common in so-called survey research, which tends to use self-report questionnaires. It is important that investigators use instruments that are reliable and valid for the population and purpose for which they will be used. Standardized instruments have manuals that provide norms and indexes of reliability and validity. However, if the populations and purpose on which these data are based are different from yours, it may be necessary for you to develop your own instrument or provide new evidence of reliability and validity. PMID- 11501699 TI - Genetics of childhood disorders: XXIX. Autoimmune disorders, part 2: Molecular mimicry. PMID- 11501700 TI - Impact of measurement uncertainty in chemical quantities on environmental prognosis by geochemical transport modelling. AB - In Germany, geochemical modelling takes a strong position in two aspects of broader public interest. The first aspect is the safety assessment of a nuclear waste repository, the second is remediation of uranium mining areas. In both aspects, the application of geochemical modelling is stipulated by authorities. This situation results from the possibility to model highly complex situations by computers. The increase in computing power experienced in recent times now offers techniques to assess the sensitivity of modelling results to uncertain input data both in the thermodynamic data base and the site-specific field data. Both aspects are investigated by using Monte Carlo methods in combination with non parametric statistics. A probabilistic geochemical modelling of uranium mill tailings leaching is demonstrated by application of TReaC modelling code using a simplified site model. PMID- 11501702 TI - Mobilization and retardation of uranium DOC species at three mine piles in Schlema/Alberoda, Saxony, Germany. AB - The modeling of geo-chemical processes needs the detailed and comprehensive knowledge of all chemical interactions existing in the flow path of waste dumps, subsoil and aquifer. This includes the adsorption, displacement and transport of heavy metal species of fulvic and humic acids, which represent the main amount of DOC in the liquid/solid system of the flow path. Comparative measurements of DOC concentrations in the input and output flow at the three waste dumps in the district Schlema/Alberoda indicated that DOC is produced and/or supplied within the waste dumps. The speciation of heavy metal compounds of fulvic and humic acids was achieved by means of sequential chromatographic analysis SCA and isoelectric focusing IEF. Between 5% and 20% of uranium exist in the flow path in the form of stable fulvic and humic acid species. Their distribution coefficients are strongly correlated with the pH values of the geo-chemical system. PMID- 11501701 TI - Environmental isotopes as a useful tool for studies at mixed uranium mill tailings sites. AB - Groundwaters in the area of a mixed landfill (domestic waste above uranium mill tailings) in Dresden (Saxony, Germany) were investigated for their isotope signatures to distinguish between different groundwater types. To determine between the two contamination sources (waste and uranium mill tailings) a multi parameter interpretation was done using both, the main hydrochemical parameters the radionuclides 234U, 238U, 226Ra and 222Rn as well as the environmental isotopes of the elements hydrogen, oxygen, sulphur and carbon. The seepage water from the landfill shows higher delta34S, delta18O and tritium values as the inflow. The tritium values give an idea about water movement in the dump and mean residence time of the groundwater. The water in the dump shows varying delta13C values which indicate different processes occurring in the dump. PMID- 11501703 TI - The mechanism of detection of air pollution by an ionization chamber. AB - The mechanism of detection of chemical vapors in air by an ionization chamber supplied by DC and AC voltage has been described. The theoretical explanation is based on numerical solutions of the differential equations of the cylindrical ionization chamber. The current of the ionization chamber operating in the AC regime has two components: a conductive component, caused by the ions drifts, and a capacitive component, caused by the distortion of the electric field. The ionization chamber operating in the DC regime has only the first component; hence the AC supplied chamber has larger response than the DC supplied chamber. PMID- 11501704 TI - Stable isotope ratios and uric acid preservation in termites belonging to three feeding habits in Thailand. AB - Nitrogen and carbon stable isotope ratios and uric acid concentrations in termites sampled from a dry evergreen forest in Thailand, were determined across three kinds of feeding habits. Feeding habits of Microcerotermes crassus, which is an abundant wood-feeder, and Dicuspiditermes makhamensis, a common soil feeding termite, were confirmed by isotopic signatures. Lichen feeding termites (Hospitalitermes birmanicus, H. bicolor and H. ataramensis) were characterized by low delta15N values, suggesting that they assimilated nitrogen deposited from the atmosphere. There was also a significant difference in uric acid concentrations between termites representing different feeding habits. No significant relationships were found between uric acid concentrations and delta15N or delta13C in Hospitalitermes. However, delta15N values were correlated with C/N ratios in H. birmanicus, except in one colony of H. ataramensis. delta13C values in both species were negatively correlated with C/N ratios. PMID- 11501705 TI - Estimating spatial and seasonal phytoplankton delta13C variations in an estuarine mangrove ecosystem. AB - In estuarine ecosystems, large spatial and seasonal variations in delta13C values of primary producers can occur, and knowledge of these variations may be crucial when interpreting stable isotope data of higher trophic levels. Obtaining clean phytoplankton samples for isotope analysis is usually impossible in such systems, and analysis of total suspended matter is not a simple proxy for phytoplankton delta13C variations. Based on a few simple assumptions regarding the C and N content of the two end-members (terrestrial detritus and phytoplankton) and the delta13C of the terrestrial component, we here present a simple model to estimate the phytoplankton delta13C variations using an existing dataset on the delta13C and elemental (C:N) composition of suspended organic matter from an estuarine mangrove ecosystem in southeast India. These variations are related to the monthly rainfall pattern during the sampling period. It is stressed that this method estimates approximate phytoplankton delta13C values, which should not be used in e.g., mixing models. However, we propose that in cases where sufficiently large datasets are available, the described procedure could provide a valuable method to semi-quantitatively estimate the seasonal or spatial variations of the phytoplankton delta13C signal. PMID- 11501706 TI - Do stable isotopes reflect the food web development in regenerating ecosystems? AB - We evaluated the use of delta15N- and delta13C-values to monitor the development of food web complexity and biodiversity in a regenerating ecosystem. Therefore a model food chain was established feeding cultivated woodlice (Porcellio dilatatus) on a cellulolytic fungus (Chaetomium globosum) grown on cellulose paper. Two diets of different quality (C:N ratios of 54 vs. 200) with different delta15N- (1.3% vs. 3.1%) but identical delta13C-values caused low and high dietary stress in animals of treatment A and B, respectively. After an incubation time of 7 weeks amount, elemental and isotopic composition of collected faeces and exuviae as well as woodlice and remaining food were determined. The increase of delta15N-values of woodlice relative to the diet was 5.7% and 2.5% in treatments A and B, respectively, whereas delta13C-shifts were 1.0% and 1.6%, showing a reverse relationship. Modelling of elemental and isotopic mass balances indicated that faeces recycling explains the unexpected high 15N-enrichments. Moreover, 13C-enrichments were positively correlated to the degree of starvation. Considering the effects of starvation and recycling of faeces, stable isotopes represent a useful tool to elucidate trophic interactions in regenerating food webs. PMID- 11501707 TI - There is no temperature dependence of net biochemical fractionation of hydrogen and oxygen isotopes in tree-ring cellulose. AB - The isotopic composition of tree-ring cellulose was obtained over a two-year period from small diameter, riparian zone trees along an elevational transect in Big Cottonwood Canyon, Utah, USA to test for a possible temperature dependence of net biological fractionation during cellulose synthesis. The isotope ratios of stream water varied by only 3.6% and 0.2% in deltaD and delta18O, respectively, over an elevation change of 810m. The similarity in stream water and macroenvironment over the short (13km) transect produced nearly constant stem and leaf water deltaD and delta18O values. In addition, what few seasonal variations observed in the isotopic composition of source water and atmospheric water vapor or in leaf water evaporative enrichment were experienced equally by all sites along the elevational transect. The temperature at each site along the transect spanned a range of > or = 5 degrees C as calculated using the adiabatic lapse rate. Since the deltaD and delta18O values of stem and leaf water varied little for these trees over this elevation/temperature transect, any differences in tree ring cellulose deltaD and delta18O values should have been associated with temperature effects on net biological fractionation. However, the slopes of the regressions of elevation versus the deltaD and delta18O values of tree-ring cellulose were not significantly different from zero indicating little or no temperature dependence of net biological fractionation. Therefore, cross-site climatic reconstruction studies using the isotope ratios of cellulose need not be concerned that temperatures during the growing season have influenced results. PMID- 11501708 TI - Continuing Medical Education and Continuing Professional Development: a credit system for monitoring and promoting excellence. AB - Probably the most important demand on the career of a medical specialist is that of having to keep up-to-date both scientifically and professionally. But the onus does not fall only on the practitioner. The institutions involved in medical teaching and professional development also have a crucial role to play by providing opportunities for continuing education and assuring that the specialist carries out enough relevant, experience-enhancing tasks to ensure continuous professional growth. As upgrading medical knowledge and developing professionally is a life-long task, both the need and the obligation to learn and improve apply to doctors of all ages and at all hierarchical levels. PMID- 11501709 TI - Another anti-allergic mechanism: antibody IgE deglycosylation induced by a substance extracted from human urine. AB - Enzymically-deglycosylated antibody IgE lost its allergic activity in mouse systemic anaphylaxis, though the IgE kept its antibody activity. IgE antibody obtained from mice treated with a substance extracted from human urine was deglycosylated. This IgE also lost the allergic activity on the systemic anaphylaxis but kept its antibody activity. These findings strongly suggest that glycosylation of IgE has a close relation to the binding of the Fc receptor and that humans have another antiallergic mechanism: in vivo IgE antibody deglycosylation induced by the substance. PMID- 11501710 TI - Epidemiology of hyperuricemia in the elderly. AB - BACKGROUND: Our study used data collected in Chung-Hsing Village to evaluate the relationship between hyperuricemia and the cardiovascular risk factors and the socio-demographic factors in the elderly in May 1998. METHODS: All individuals aged 65 and over were studied. A total of 1093 subjects, out of 1774 registered residents, were contacted by face-to-face interview. The response rate was 61.6 percent. However only 586 respondents had blood tests and completed questionnaires. Analysis in this study was based on these 586 subjects. To study the significant correlates of hyperuricemia, t- test, two-way ANOVA, chi-square test and multivariate logistic regression were used. RESULTS: Our results showed that 66 percent were men and 34 percent were women. The mean age was 73.1+/-5.3 years. The proportions of hyperuricemia were 57.3 percent in men and 40.9 percent in women (p < .01). In chi-square test, hypercholesterolemia, hypertriglyceridemia, renal function impairment, retirement status and marital status were related to hyperuricemia. After controlling the other covariates, the multivariate logistic regression analysis showed that the significant related factors of hyperuricemia were hypercholesterolemia, hypertriglyceridemia, and renal function impairment. CONCLUSIONS: Hyperuricemia is often found in the elderly. Hyperuricemia is more common in elderly men than in elderly women. A large-scale investigation will be suggested in the future to address causal effect issues between hyperuricemia and hypercholesterolemia, hypertriglyceridemia, or renalfunction impairment. PMID- 11501711 TI - A case of pleural mesothelioma with effusive-constrictive pericarditis. PMID- 11501712 TI - Sweet's syndrome in a patient with acute ulcerative colitis: presentation of a case and review of the literature. PMID- 11501713 TI - The second sacrifice: costly advances in medicine and surgery during the Civil War. PMID- 11501714 TI - Peculiar elongated and sickle-shaped red blood corpuscles in a case of severe anemia. 1910. PMID- 11501715 TI - Anatomy and Leonardo da Vinci. PMID- 11501716 TI - Biotechnology at Yale. PMID- 11501717 TI - Biopsychosocial mediators and moderators of stress-health relationships in patients with recently diagnosed rheumatoid arthritis. AB - OBJECTIVE: To investigate the mediating and moderating roles of social support, coping, and physiological variables in the relationship between life events and health status. METHODS: Psychological and biological measurements were taken in 54 patients (38 women, 16 men, mean age +/- SD 56 +/-14.4 years) with recently diagnosed rheumatoid arthritis (RA). RESULTS: Life events were correlated with psychological distress, but not with disease activity. No mediators for the relationship of life events with psychological well-being and disease activity were observed. In 40 tests, 4 moderators were found: Problem-focused coping, perceived support, diastolic blood pressure, and total number of lymphocytes were moderators of the relationship between daily hassles and health status (P < or = 0.05). CONCLUSION: Our study provides limited support for the notion that the interactions of life stress with biopsychosocial variables have an impact on health. None of these variables were found to be crucial mediators of stress health associations in recently diagnosed patients with RA, but some provocative evidence was given that biopsychosocial variables may have a minor impact on stress-health relationships. PMID- 11501718 TI - Development of an instrument to measure pain in rheumatoid arthritis: Rheumatoid Arthritis Pain Scale (RAPS). AB - OBJECTIVE: To develop a valid and reliable clinical instrument for measuring pain in adult patients with rheumatoid arthritis. The resulting Rheumatoid Arthritis Pain Scale (RAPS) is a quantitative, single-score, self-report 24-item instrument. METHODS: Psychometric evaluation of RAPS was conducted following estimation of content validity and a pilot study. The actual study used a convenience sample of 120 adults, 18 years of age or older, with pain of at least 3 months duration. The setting was a large rheumatology private practice in a metropolitan southwestern city. The gate control and affective motivational theories of pain served as a framework guiding the development of RAPS, which includes items suggestive of the multidimensional pain experience in rheumatoid arthritis. Four subscales, physiological, affective, sensory-discriminative, and cognitive, evaluated numerous pain factors. RESULTS: Findings indicate a high estimate for internal consistency for the total scale and a moderate to high estimate of internal consistency for projected subscales. Data were analyzed using Cronbach's coefficient alpha, Pearson product-moment correlation coefficients, and exploratory factor analysis. Using Cronbach's coefficient alpha, RAPS showed an internal consistency reliability coefficient of 0.92, a strong indicator of reliability. Reliability assessments for the 4 subscales also indicate reliability, with Cronbach's coefficients ranging from 0.65 to 0.86. Exploratory factor analysis yielded 3 factors with criteria for factor loadings > or = 0.4. CONCLUSION: The study's findings provided support for RAPS as a reliable and valid measurement of rheumatoid arthritis pain. Assessment of rheumatoid arthritis pain and its relationship to treatment outcomes could significantly impact the treatment interventions. PMID- 11501719 TI - Health care utilization among rheumatoid arthritis patients referred to a rheumatology center: unequal needs, unequal care? AB - OBJECTIVE: To quantify the utilization of health care by rheumatoid arthritis (RA) patients and to estimate the contribution of patient characteristics to the explanation of the use of care, in order to evaluate whether those in need of care actually receive care. METHODS: A questionnaire survey and a clinical examination were conducted among patients with RA referred to a rheumatology center. Health care utilization was assessed for medical care, allied health care, psychosocial care, and home care. The influence of sociodemographic variables and clinical and health characteristics on health care utilization was assessed by means of logistic regression. RESULTS: Multivariate analyses showed that, for all types of services, disease-related factors explained most of the utilization. However, some sociodemographic variables (age, sex, and living situation) were also related to the utilization of care. CONCLUSION: Most patients received the care they needed. However, for the elderly with RA, problems in access to allied health care and psychosocial care exist. PMID- 11501720 TI - Modifiers of change in physical functioning in older adults with knee pain: the Observational Arthritis Study in Seniors (OASIS). AB - OBJECTIVE: To ascertain predictors of decline in physical functioning among older adults reporting knee pain. METHODS: The Observational Arthritis Study in Seniors was a longitudinal study of 480 adults over 65 years of age. Measurements of strength, sociodemographic characteristics, disease burden (including radiographic knee osteoarthritis [OA]), self-reported disability, and functional limitations were obtained on participants at baseline and at 15 and 30 months. RESULTS: Radiographic evidence of OA at baseline was moderately associated with an increased decline in both transfer (P = 0.06) and ambulatory-based performance tasks (P = 0.04) but not in self-reported disability. This effect disappeared after accounting for baseline levels of knee pain intensity and knee strength. CONCLUSION: Knee pain intensity and knee strength may mediate the relationship between radiographic evidence of knee OA and change in performance. Although it is not clear whether joint disease precedes or follows a decline in muscular strength, these results may help to identify subpopulations of older persons with knee OA who may benefit from interventions aimed at slowing the progression of disability related to transfer and ambulatory-based tasks. PMID- 11501721 TI - Variation in perceptions of treatment and self-care practices in elderly with osteoarthritis: a comparison between African American and white patients. AB - OBJECTIVE: To compare elderly African American and white patients with osteoarthritis of the knee or hip with respect to their perceptions of the efficacy of traditional and complementary treatments and their self-care practices. METHODS: An observational, cross-sectional study design using structured questionnaires was employed. RESULTS: The sample consisted of 593 patients (44% African American and 56% white). The 2 groups were comparable with respect to age, disease severity or functional status, and comorbidities. African Americans were more likely than whites to report lower educational level and household income. African Americans were also more likely than whites to perceive various traditional and complementary care modalities as efficacious. However, they were less likely than whites to perceive joint replacement therapy as efficacious (odds ratio 0.52, 95% confidence interval 0.28-0.98). African American patients were more likely than white patients to rely on self-care measures for their arthritis. CONCLUSION: African American and white patients with osteoarthritis of the knee or hip differ with respect to their perceptions of traditional and complementary treatments for arthritis and their self-care practices. PMID- 11501722 TI - Esophageal involvement and pulmonary manifestations in systemic sclerosis. AB - OBJECTIVE: To assess whether esophageal manometric motor disturbances are associated with abnormalities consistent with interstitial lung disease (ILD) on both pulmonary function tests (PFT) and high resolution computerized tomography (HRCT) scans in patients with systemic sclerosis (SSc), during initial evaluation and at 2 years followup. METHODS: Esophageal manometry, PFT, and HRCT scans were performed in 43 consecutive SSc patients. PFT and HRCT scan parameters were compared between patients with severe esophageal motor dysfunction (i.e., aperistalsis and decreased low esophageal sphincter pressure), patients with moderate esophageal motor dysfunction (hypoperistalsis), and patients without esophageal motor dysfunction on manometry. RESULTS: During initial evaluation of SSc, patients with severe esophageal motor impairment, compared with those with moderate and without esophageal dysmotility, exhibited significantly decreased median values of diffusing capacity for carbon monoxide (DLco) (68% vs. 94% vs. 104%) and exhibited higher prevalence of evidence for ILD on HRCT scan (57% vs. 27% vs. 18%). At 2 years followup of SSc, patients with severe esophageal motor disturbances, compared with those without, had faster deterioration of DLco median values (-16% vs. +1%) and higher frequency of ILD on HRCT scan (70% vs. 25%). CONCLUSION: Our series underscores a correlation between the degree of esophageal manometric motor disturbances and evidence for ILD on PFT and HRCT scan in SSc patients, suggesting that gastroesophageal reflux (GER) may be one of the contributing factors of ILD in SSc. Our findings further indicate that patients with severe esophageal impairment may require closer followup of lung parameters. In turn, it suggests that aggressive therapy of GER should be initiated in these SSc patients, as it may result in decreased deterioration of pulmonary function. PMID- 11501723 TI - Pain and pain relief in fibromyalgia patients followed for three years. AB - OBJECTIVE: To examine the natural clinical course of pain in fibromyalgia (FM) and patients' reports of the use of interventions for pain relief. METHODS: This prospective 3-year study examined pain, and the treatment thereof, in a cohort of 82 women with FM, of whom 59 (72%) were reassessed on 3 subsequent occasions. Pain was measured by the following parameters: visual analog scale (VASpain), tender point count (TP), and the occurrence of widespread pain (WP). Function was assessed by the Health Assessment Questionnaire and the Fibromyalgia Impact Questionnaire, and depression and anxiety by the Arthritis Impact Measurement Scales. All treatments for FM were recorded, and patients identified the treatment that they believed had helped their symptoms of FM. RESULTS: Pain reporting as measured by all parameters decreased significantly for the whole group over the duration of the study. The mean VASpain decreased from 66 to 55, the mean TP count decreased from 13.5 to 10.5, and the number of patients with WP decreased from 100% to 63%. VASpain correlated positively with TP and WP. One third of patients experienced a reduction in pain by at least 30% from baseline as well as a better outcome in overall status of FM. There was a decline in the use of prescribed medications, whereas the use of alternative products increased. Physical treatment modalities were more often perceived to be of benefit than prescribed medications. CONCLUSION: We have observed a spontaneous improvement in pain reporting and less medication use in FM patients, suggesting that the course of this condition may be more favorable than has previously been reported. PMID- 11501724 TI - Psychosocial and health status variables independently predict health care seeking in fibromyalgia. AB - OBJECTIVE: To determine whether variables derived from the self-regulatory model of health and illness behavior accurately predict status as a patient or nonpatient with fibromyalgia (FM). METHODS: Subjects were 79 patients who met American College of Rheumatology (ACR) criteria for FM and 39 community residents who met ACR criteria for FM but had not sought medical care for their symptoms (nonpatients). Subjects were administered 14 measures that produced 6 domains of variables: background demographics and pain duration; psychiatric morbidity; and personality, environmental, cognitive, and health status factors. These domains were entered in 4 different hierarchical logistic regression analyses to predict status as patient or nonpatient. RESULTS: The full regression model was statistically significant (P < 0.0001) and correctly identified 90.7% of the subjects with a sensitivity of 92.4% and a specificity of 87.2%. The best individual predictors of group status were self-reports of self-efficacy, negative affect, recent stressful events, and perceived pain. Relative to nonpatients, patients reported higher levels of negative affect and perceived pain and a greater number of recent stressful experiences, as well as lower levels of self-efficacy. CONCLUSION: Consistent with the self-regulatory model of health and illness behavior, psychosocial and health status variables predict health care-seeking behavior in persons with FM independently of background demographics and psychiatric morbidity. These variables may influence the severity of symptoms experienced by persons with this disorder as well as their health care-seeking behavior, but they are not necessary to produce abnormal pain sensitivity in FM. PMID- 11501725 TI - Rheumatology patient and physician concordance with respect to important health and symptom status outcomes. AB - OBJECTIVE: In chronic diseases such as arthritis, patients' priorities and values are of prime importance in the measurement of outcome. Relatively few indices take this into account. We sought to identify individual rheumatology patients' choices for important health and symptom status outcomes and assess physicians' ability to detect them. METHODS: A cross-sectional, observational study design was used to examine rheumatology patients' choices for important health and symptom status outcomes. A descriptive summary of patients' choices of "most important" health and symptom status outcomes was compiled. The sensitivity and specificity of physicians' ability to detect their patients' choices for important health or symptom status outcomes were calculated. RESULTS: The sample consisted of 79 rheumatology patients, 30 men and 49 women, with a mean age of 52.6 years. The feeling of being in control was the mental health outcome rated most important by the largest number of the patients (42%); cognitive function (making decisions, solving problems) (22%), feelings of being happy (18%), and feelings of being satisfied (16%) were also rated as being "most important" by many patients. Feelings of hope were rated "most important" by the fewest patients (2%). The physicians' sensitivity to detecting patients' choices of most important outcomes was highest for social functioning and lowest for mental functioning. The specificity to detect patients' choices of less important outcomes was highest for physical functioning and lowest for symptom status. CONCLUSION: In this study involving 4 rheumatologists, patients were found to differ in their choices of important health and symptom status outcomes. Physicians often fail to identify these choices. PMID- 11501726 TI - Volunteers' experiences of becoming arthritis self-management lay leaders: "It's almost as if I've stopped aging and started to get younger!". AB - OBJECTIVE: To determine whether undergoing training to become a lay leader and conducting an arthritis self-management course is associated with improvements in physical and psychological health status, arthritis self-efficacy, use of self management techniques, and visits to the general practitioner. In addition, we aimed to describe the experiences of training and course delivery from the older volunteers' perspective. METHODS: 21 participants completed all assessments and had a median age of 58, median disease duration of 10 years, and either osteoarthritis (n = 13) or rheumatoid arthritis (n = 8). The study was a pretest posttest design with qualitative data collected at 3 points in time: before training, 6 weeks after training, and 6 months after training. Quantitative data were collected through self-administered postal questionnaires at baseline and 6 month followup. RESULT: Six months after training, participants reported small, significant increases in arthritis self-efficacy for pain (P = 0.002), cognitive symptom management (P = 0.004), and communication with their physician (P = 0.024) and a small, significant decrease in depressed mood (P = 0.04). Qualitative data supported these findings, with participants reporting more confidence, happiness, and a changed outlook on life in general. Volunteerism was associated with altruistic behavior and with filling the vocational void caused by retirement. CONCLUSION: Findings support the value of volunteerism and training to become lay leaders in arthritis self-management programs. Volunteers reported positive changes both in themselves and in course participants. They enjoyed helping similar others and being involved in a worthwhile activity, and they valued their newly acquired status as lay leaders. Many had begun to apply their newfound knowledge about self-management to their own situation, reporting less pain and more willingness "to get on with life." PMID- 11501727 TI - Smallest detectable and minimal clinically important differences of rehabilitation intervention with their implications for required sample sizes using WOMAC and SF-36 quality of life measurement instruments in patients with osteoarthritis of the lower extremities. AB - OBJECTIVE: To discuss the concepts of the minimal clinically important difference (MCID) and the smallest detectable difference (SDD) and to examine their relation to required sample sizes for future studies using concrete data of the condition specific Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the generic Medical Outcomes Study 36-Item Short Form (SF-36) in patients with osteoarthritis of the lower extremities undergoing a comprehensive inpatient rehabilitation intervention. METHODS: SDD and MCID were determined in a prospective study of 122 patients before a comprehensive inpatient rehabilitation intervention and at the 3-month followup. MCID was assessed by the transition method. Required SDD and sample sizes were determined by applying normal approximation and taking into account the calculation of power. RESULTS: In the WOMAC sections the SDD and MCID ranged from 0.51 to 1.33 points (scale 0 to 10), and in the SF-36 sections the SDD and MCID ranged from 2.0 to 7.8 points (scale 0 to 100). Both questionnaires showed 2 moderately responsive sections that led to required sample sizes of 40 to 325 per treatment arm for a clinical study with unpaired data or total for paired followup data. CONCLUSION: In rehabilitation intervention, effects larger than 12% of baseline score (6% of maximal score) can be attained and detected as MCID by the transition method in both the WOMAC and the SF-36. Effects of this size lead to reasonable sample sizes for future studies lying below n = 300. The same holds true for moderately responsive questionnaire sections with effect sizes higher than 0.25. When designing studies, assumed effects below the MCID may be detectable but are clinically meaningless. PMID- 11501728 TI - Physical and exercise therapy for treatment of the rheumatoid hand. AB - OBJECTIVE: To study the short-term effects of physical therapy (ice massage or wax packs, thermal baths, and faradic hand baths) and exercise therapy on the rheumatoid hand. METHODS: The effect of individual physical therapy and exercise therapy programs was evaluated in 50 randomly selected rheumatoid arthritis inpatients (38 women and 12 men). Mean patient age (+/- SD) was 47.94 +/- 11.22 years, and mean disease duration was 5.04 +/- 4.80 years. The control group consisted of 50 randomly selected rheumatoid arthritis outpatients (37 women and 13 men; mean age 48.46 +/- 10.65 years, mean duration of disease 5.23 +/- 4.89 years) who at the time of the investigation were not receiving any physical or exercise therapy. The clinical indices used for evaluation of inflammation included erythrocyte sedimentation rate (ESR), pain intensity, proximal interphalangeal (PIP) joint size, and Ritchie articular index. Hand grip strength, palmar tip-to-tip and key pinch finger strength, finger range of motion, and activities of daily living (ADL) were the parameters used to assess the functional hand status. The study was single-blinded and of 3 weeks duration. RESULTS: In the physical therapy treated group, there was an improvement for most of the observed indices from baseline parameters that achieved statistical significance (P < 0.01 and P < 0.005) after the 3-week study period. ESR and PIP joint size improved clinically but failed to reach statistical significance. Patients had a more significant improvement in hand pain, joint tenderness, and ADL score (P < 0.005) than in range of motion (P < 0.01). All parameters in the control group slightly deteriorated over the study period. CONCLUSION: At least in the short term, physical and, particularly, exercise therapy produce a favorable improvement in the functional status of the rheumatoid hand. PMID- 11501729 TI - Use of complementary and alternative medicine by older patients with arthritis: a population-based study. AB - OBJECTIVE: To determine the prevalence of complementary and alternative medicine (CAM) use and to identify factors associated with its use in older patients with arthritis. METHODS: A population-based telephone survey of 480 elderly patients with arthritis was conducted to determine demographics, comorbidities, health status, arthritis symptoms, and the use of CAM and traditional providers and treatments for arthritis. RESULTS: CAM provider use was reported by 28% of respondents, and 66% reported using one or more CAM treatments. Factors independently related to CAM provider use (P < 0.05) included podiatrist or orthotist use, physician visits for arthritis, and fair or poor self-reported health. For CAM treatments, independent associations were found with physical or occupational therapist use, physician visits for arthritis, chronic obstructive pulmonary disease, and alcohol abstinence. Rural residence, age, income, education, and health insurance type were unrelated to CAM use. CONCLUSION: Many older patients with arthritis reported seeing CAM providers, and most used CAM treatments. The use of CAM for arthritis was most common among those with poorer self-assessed health and higher use of traditional health care resources. PMID- 11501730 TI - Rapid remission of treatment-resistant ankylosing spondylitis with etanercept--a drug for refractory ankylosing spondylitis? PMID- 11501731 TI - Functional neuroimaging of cognition: state-of-the-art. AB - This paper presents a brief overview of functional neuroimaging studies of cognitive processes. A "publication profile" is identified for PET and fMRI studies of cognitive functions. It is shown that the number of publications has increased exponentially over the last ten years, and that a set of ten journals accounts for the vast majority of publications. It is argued that the field has moved from a state of explorative character to a state of high theoretical sophistication. With special reference to the contributions in this issue of Scandinavian Journal of Psychology, methodological, topical, regional, and clinical implications of cognitive neuroscientific studies are discussed. PMID- 11501732 TI - Generative models, brain function and neuroimaging. AB - The representational capacity and inherent function of any neuron, neuronal population or cortical area in the brain is dynamic and context-sensitive. Functional integration, or interactions among brain systems, that employ driving (bottom up) and backward (top-down) connections, mediate this adaptive and contextual specialisation. A critical consequence is that neuronal responses, in any given cortical area, can represent different things at different times. This can have fundamental implications for the design of brain imaging experiments and the interpretation of their results. Our arguments are developed under generative models of brain function, where higher-level systems provide a prediction of the inputs to lower-level regions. Conflict between the two is resolved by changes in the higher-level representations, which are driven by the ensuing error in lower regions, until the mismatch is "cancelled". From this perspective the specialisation of any region is determined both by bottom-up driving inputs and by top-down predictions. Specialisation is therefore not an intrinsic property of any region but depends on both forward and backward connections with other areas. Because the latter have access to the context in which the inputs are generated they are in a position to modulate the selectivity or specialisation of lower areas. The implications for classical models (e.g., classical receptive fields in electrophysiology, classical specialisation in neuroimaging and connectionism in cognitive models) are severe and suggest these models may provide incomplete accounts of real brain architectures. Here we focus on the implications for cognitive neuroscience in the context of neuroimaging. PMID- 11501733 TI - Repetition effects for words and nonwords as indexed by event-related fMRI: a preliminary study. AB - We have previously shown differential effects of stimulus familiarity on the repetition-related responses in right fusiform cortex to both faces and symbols. Repetition of familiar stimuli produced a response decrease, whereas repetition of unfamiliar stimuli produced a response increase. In the present experiment, we used words and nonwords as the familiar and unfamiliar stimuli respectively. In this case, the only fusiform region showing the familiarity-by-repetition interaction was in anterior left fusiform. This left-lateralisation of the fusiform interaction is consistent with our hypothesis that these repetition related effects occur in the same regions responsible for perceptual recognition of familiar stimuli. PMID- 11501734 TI - On the relation between conceptual priming, neural priming, and novelty assessment. AB - A consistently reported finding in functional neuroimaging studies which compare processing of new information to processing of old information is a reduction in blood flow, and hence neural activity, associated with the old condition. This deactivation has been labeled neural priming. Some investigators have hypothesized that neural priming is the physiological mechanism underlying conceptual priming--a facilitation in the semantic processing of repeated information. Others, however, have hypothesized that neural priming reflects novelty assessment--a mechanism which minimizes the probability that redundant information will be stored in long-term memory. In this paper, the conceptual priming and novelty assessment hypotheses are compared and contrasted in order to ask, and tentatively answer, the question: Are conceptual priming and novelty assessment cognitively and neurophysiologically distinct? Based on a review of the literature, it is suggested that whereas novelty assessment and conceptual priming are distinct cognitive entities, they cannot be presently separated neurophysiologically. That is, some novelty assessment deactivations may in fact reflect priming, and some priming deactivations may in fact reflect novelty assessment. PMID- 11501735 TI - Learning related modulation of functional retrieval networks in man. AB - The medial temporal lobe has been implicated in studies of episodic memory tasks involving spatio-temporal context and object-location conjunctions. We have previously demonstrated that an increased level of practice in a free-recall task parallels a decrease in the functional activity of several brain regions, including the medial temporal lobe, the prefrontal, the anterior cingulate, the anterior insular, and the posterior parietal cortices, that in concert demonstrate a move from elaborate controlled processing towards a higher degree of automaticity. Here we report data from two experiments that extend these initial observations. We used a similar experimental approach but probed for effects of retrieval paradigms and stimulus material. In the first experiment we investigated practice related changes during recognition of object-location conjunctions and in the second during free-recall of pseudo-words. Learning in a neural network is a dynamic consequence of information processing and network plasticity. The present and previous PET results indicate that practice can induce a learning related functional restructuring of information processing. Different adaptive processes likely subserve the functional re-organisation observed. These may in part be related to different demands for attentional and working memory processing. It appears that the role(s) of the prefrontal cortex and the medial temporal lobe in memory retrieval are complex, perhaps reflecting several different interacting processes or cognitive components. We suggest that an integrative interactive perspective on the role of the prefrontal and medial temporal lobe is necessary for an understanding of the processing significance of these regions in learning and memory. It appears necessary to develop elaborated and explicit computational models for prefrontal and medial temporal functions in order to derive detailed empirical predictions, and in combination with an efficient use and development of functional neuroimaging approaches, to further the understanding of the processing significance of these regions in memory. PMID- 11501736 TI - The role of spatial configuration in tests of working memory explored with functional neuroimaging. AB - While the importance of the prefrontal cortex for "higher-order" cognitive functions is largely undisputed, no consensus has been reached regarding the fractionation of functions within this region. Several recent functional neuroimaging studies have suggested that the mid-ventrolateral frontal cortex may play an important role in various aspects of human memory. Thus, similar patterns of activation have been observed in this region during analogous spatial, verbal and visual span tasks. In the present study, however, activation was observed in a more dorsolateral region of the lateral frontal cortex during a modified version of the spatial span task, which differed only in the spatial configuration of the array employed. The results of a supplementary behavioral study, designed to investigate this effect further, suggest that in spatial memory tasks certain stimulus configurations may encourage subjects to adopt mnemonic strategies, which may depend upon dorsolateral, rather than ventrolateral, regions of the frontal cortex. These findings shed further light on the functional relationship between dorsal and ventral regions of the lateral frontal cortex and, more specifically, how the "executive" processes assumed to be dependent upon these regions might contribute to aspects of human memory. PMID- 11501737 TI - The retrosplenial contribution to human navigation: a review of lesion and neuroimaging findings. AB - The clinical and neuroimaging literatures are surveyed in order to collate for the first time the available data on retrosplenial involvement in human navigation. Several notable features emerge from consideration of the case reports of relatively pure topographical disorientation in the presence of a retrosplenial lesion. The majority of cases follow damage to the right retrosplenial cortex, with Brodmann's area 30 apparently compromised in most cases. All patients displayed impaired learning of new routes, and defective navigation in familiar environments complaining they could not use preserved landmark recognition to aid orientation. The deficit generally resolved within eight weeks of onset. The majority of functional neuroimaging studies involving navigation or orientation in large-scale space also activate the retrosplenial cortex, usually bilaterally, with good concordance in the locations of the voxel of peak activation across studies, again with Brodmann's area 30 featuring prominently. While there is strong evidence for right medial temporal lobe involvement in navigation, it now seems that the inputs the hippocampus and related structures receive from and convey to right retrosplenial cortex have a similar spatial preference, while the left medial temporal and left retrosplenial cortices seem primarily concerned with more general aspects of episodic memory. PMID- 11501738 TI - Neuroimaging of declarative memory in schizophrenia. AB - The past three decades have seen tremendous growth in our understanding of the cerebral underpinnings of schizophrenia. including the neural correlates of the cognitive impairment seen in this syndrome. In this article we review the role that structural and functional neuroimaging has played in elucidating the cerebral basis for the declarative memory deficits associated with schizophrenia. Memory impairment in schizophrenia appears to involve abnormal connectivity between the prefrontal cortex and three regions important in normal learning and memory: the hippocampus, thalamus, and cerebellum. PMID- 11501739 TI - Cognitive processing in literate and illiterate subjects: a review of some recent behavioral and functional neuroimaging data. AB - The study of illiterate subjects, which for specific socio-cultural reasons did not have the opportunity to acquire basic reading and writing skills, represents one approach to study the interaction between neurobiological and cultural factors in cognitive development and the functional organization of the human brain. In addition the naturally occurring illiteracy may serve as a model for studying the influence of alphabetic orthography on auditory-verbal language. In this paper we have reviewed some recent behavioral and functional neuroimaging data indicating that learning an alphabetic written language modulates the auditory-verbal language system in a non-trivial way and provided support for the hypothesis that the functional architecture of the brain is modulated by literacy. We have also indicated that the effects of literacy and formal schooling is not limited to language related skills but appears to affect also other cognitive domains. In particular, we indicate that formal schooling influences 2D but not 3D visual naming skills. We have also pointed to the importance of using ecologically relevant tasks when comparing literate and illiterate subjects. We also demonstrate the applicability of a network approach in elucidating differences in the functional organization of the brain between groups. The strength of such an approach is the ability to study patterns of interactions between functionally specialized brain regions and the possibility to compare such patterns of brain interactions between groups or functional states. This complements the more commonly used activation approach to functional neuroimaging data, which characterize functionally specialized regions, and provides important data characterizing the functional interactions between these regions. PMID- 11501740 TI - Common pathways in mental imagery and pain perception: an fMRI study of a subject with an amputated arm. AB - The present paper reviews data from two previous studies in our laboratory, as well as some additional new data, on the neuronal representation of movement and pain imagery in a subject with an amputated right arm. The subject imagined painful and non-painful finger movements in the amputated stump while being in a MRI scanner, acquiring EPI-images for fMRI analysis. In Study I (Ersland et al., 1996) the Subject alternated tapping with his intact left hand fingers and imagining "tapping" with the fingers of his amputated right arm. The results showed increased neuronal activation in the right motor cortex (precentral gyrus) when tapping with the fingers of the left hand, and a corresponding activation in the left motor cortex when imagining tapping with the fingers of the amputated right arm. Finger tappings of the intact left hand fingers also resulted in a larger activated precentral area than imagery "finger tapping" of the amputated right arm fingers. In Study II (Rosen et al., 2001 in press) the same subject imagining painful and pleasurable finger movements, and still positions of the fingers of the amputated arm. The results showed larger activations over the motor cortex for movement imagining versus imagining the hand being in a still position, and larger activations over the sensory cortex when imagining painful experiences. It can therefore be concluded that not only does imagery activate the same motor areas as real finger movements, but also that adding instructions of pain together with imaging moving the fingers intensified the activation compared with adding instructions about non-painful experiences. From these studies, it is clear that areas activated during actual motor execution to a large extent also are activated during mental imagery of the same motor commands. In this respect the present studies add to studies of visual imagery that have shown a similar correspondence in activation between actual object perception and imagery of the same object. PMID- 11501741 TI - Cognitive neuroscience of aging: contributions of functional neuroimaging. AB - By revealing how brain activity during cognitive performance changes as a function of aging, studies using positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) are contributing to the development of a new discipline of Cognitive Neuroscience of Aging. This article reviews functional neuroimaging studies of cognitive aging in the domains of visual perception, episodic memory encoding and semantic memory retrieval, episodic memory retrieval, implicit memory, and working memory. The most consistent finding of these studies was that brain activity tends to be less lateralized in older adults than in younger adults. This finding is conceptualized in terms of a model called Hemispheric Asymmetry Reduction in Old Adults (HAROLD). According to a compensation hypothesis, bihemispheric involvement could help counteract age related neurocognitive decline, whereas, according to a dedifferentiation hypothesis, it reflects a difficulty in recruiting specialized neural mechanisms. PMID- 11501742 TI - Dopamine and cognitive functioning: brain imaging findings in Huntington's disease and normal aging. AB - Recent brain imaging studies in Huntington's disease (HD) and normal aging suggest a relationship between central dopaminergic neurotransmission and cognitive performance. Results demonstrate substantial losses in dopamine (DA) function in both HD and aging. Moreover, HD patients and older adults show deficits across multiple cognitive domains, including episodic memory, speed of processing, and executive functioning. Although few studies are available at present, there is converging evidence that multiple measures of pre- and postsynaptic DA biochemistry are (a) highly interrelated, and (b) strongly associated with the cognitive deficits that accompany HD and aging. There is also emerging evidence that DA neurotransmission influences cognitive performance independent of HD or age. In general, the research reviewed in this article indicates that the nigrostriatal DA system is an important component of a frontostriatal circuitry that is critically involved in cognitive functioning. PMID- 11501743 TI - Transcranial magnetic stimulation as a tool for cognitive studies. AB - Transcranial Magnetic Stimulation (TMS) is a tool for the non-invasive stimulation of the human brain. It allows the activation of arbitrary sites of the superficial cortex and, combined with other brain-imaging techniques such as EEG, PET, and fMRI, it can be used to evaluate cortical excitability and connectivity. This is of major importance in, for example, the study of cognitive processes such as language, learning, memory and self-representation, which are thought to be represented in multiple brain areas. The mechanisms of action of TMS are known on a basic level, but its effect on the activation state of brain tissue is still poorly understood. Clinical applications of TMS have also been proposed and guidelines for its safe use drafted. PMID- 11501744 TI - Utilization of oncoprotein-pulsed dendritic cells as tumor vaccines. PMID- 11501745 TI - The nature of giant cell tumor of bone. AB - Giant cell tumor of bone (GCT) is a locally osteolytic tumor with variable aggressiveness. In rare cases, pulmonary metastasis can be observed. The lesion most frequently occurs in the epiphysis of long tubular bones of the knee region, predominantly affecting young adults after closure of the growth plate. The characteristic histological appearance of GCT displays a high number of osteoclast-like multinucleated giant cells, which resulted in the classification "osteoclastoma" or "giant cell tumor". Apart from the multinucleated giant cells, there are two mononuclear cell types in GCT. The first one has a round morphology and resembles monocytes. The second cell type is the spindle-shaped, fibroblast like stromal cell. Cell culture experiments with GCT cells revealed the stromal cell to be the proliferating component of the GCT. The other two cell types, the monocyte and the multinucleated giant cell, were lost after a few cell culture passages. Furthermore, latest results from GCT reveal that the stromal cells secrete a variety of cytokines and differentiation factors, including MCP1, ODF, and M-CSF. These molecules are monocyte chemoattractants and are essential for osteoclast differentiation, suggesting that the stromal cell stimulates blood monocyte immigration into tumor tissue and enhances their fusion into osteoclast like, multinucleated giant cells. The multinucleated giant cell itself resembles a normal osteoclast that is able to resorb bone leading to extended osteolysis. This new model of GCT genesis supports the hypothesis that the stromal cell is the neoplastic component whilst the monocytes and the multinucleated giant cells are just reactive components of this tumor. Taking this into consideration, the nomenclature of the "giant cell tumor" needs to be reconsidered. PMID- 11501746 TI - Treatment of early stage testicular seminoma. AB - Stage I and IIA/B testicular seminoma represent approximately 45% of all testicular germ cell tumours. Due to the availability of highly efficient salvage treatment, the disease-specific survival in stage I seminoma is approximately 100%, irrespective of the choice of adjuvant treatment. Radiotherapy with 26 Gy to the paraaortic/paracaval lymph nodes yields excellent cure rates of 95 98% with a favourable profile of acute and late toxicity. Likewise, phase-II trials with single-agent carboplatinum systemic treatment have demonstrated a rate of relapse of 3-4% on average. However, carboplatinum chemotherapy has to be regarded as experimental until data of phase-III trials are available. Surveillance in stage I disease is conflicted with a rate of relapse of approximately 20%. However, 80% of the patients will avoid potentially toxic overtreatment by the watch-and-wait policy. In stage IIA/B seminoma, "dogleg" radiotherapy with 30 Gy and 36 Gy, respectively, provides high cure rates of 90 95%. Those patients relapsing will be salvaged in almost 100% of cases. Testicular intraepithelial neoplasia (TIN) is the common precursor lesion of testicular germ cell tumours except for spermatocytic seminoma. In case of TIN in a single testis or bilateral TIN, local radiotherapy with 18 Gy is recommended as standard treatment. PMID- 11501747 TI - Chromosome 8p deletion is associated with metastasis of human hepatocellular carcinoma when high and low metastatic models are compared. AB - Recently, we found that chromosome 8p deletion might be associated with hepatocellular carcinoma (HCC) metastasis by analyzing the differences in chromosomal alterations between primary tumors and their matched metastatic lesions of HCC with comparative genomic hybridization (CGH) (Qin et al. 1999). To further confirm this interesting finding, the genomic changes of two models bearing human HCC with different metastatic potentials (LCI-D20 and LCI-D35), and the new human HCC cell line with high metastatic potential (MHCC97) were analyzed by CGH. Gains on 1q, 6q, 7p, and 8q, and losses on 13p, 14p, 19p, 21, and 22 were detected in both LCI-D20 and LCI-D35 models. However, high copy number amplification of a minimum region at 1q12-q22 and 12q, and deletions on 1p32 pter, 3p21-pter, 8p, 9p, 10q, 14q, and 15p were detected only in the LCI-D20 model. Gains on 1p21-p32, 2p13-p21, 6p12-pter, 9p, 15q, and 16q11-q21, and losses on 2p23-pter, 4q24-qter, 7q31-qter, 12q, 17p, and 18 were detected only in the LCI-D35 model. The chromosomal aberration patterns in the MHCC97 cell line were similar to its parent LCI-D20 model, except that gains on 19q and losses on 4, 5, 10q, and 13q were found only in the cell line. These results provide some indirect clues to the metastasis-related chromosomal aberrations of HCC and further support the finding that 8p deletion is associated with HCC metastasis. 1q12-22 and 12q might harbor a novel oncogene(s) that contributes to the development and progression of HCC. Amplification on 8q and deletions on 4q and 17p may be not necessary for HCC metastasis. PMID- 11501748 TI - Immunobead RT-PCR versus regular RT-PCR amplification of CEA mRNA in peripheral blood. AB - PURPOSE: The reverse transcription polymerase chain reaction (RT-PCR) amplification of tumor-specific mRNA has been used for the detection of cancer cells in peripheral blood. More recently, an immunomagnetic isolation and reverse transcription polymerase chain reaction (immunobead RT-PCR) was developed which has reportedly significant advantages over the previous RT-PCR analysis. In our study, we compared these two methods using a model set of peripheral blood containing tumor cells under standardized conditions. MATERIAL AND METHODS: In order to compare the false positive rate, normal peripheral blood samples from five volunteers were analyzed by both methods. A model set of peripheral blood containing tumor cells was established by adding SNUC4 human colon cancer cells to peripheral blood collected from normal volunteers not showing any nonspecific bands upon electrophoresis of the PCR products. RT-PCR amplification of carcinoembryonic antigen (CEA) mRNA was done with total RNA and mRNA prepared from this model sample. In immunobead RT-PCR analysis, mRNA was prepared from the cells isolated with anti-CEA antibody-coated magnetic beads or anti-Ber-EP4 antibody-coated magnetic beads before the RT-PCR analysis. RESULT: The immunobead RT-PCR yielded no non-specific band, while the regular RT-PCR using total RNA did show non-specific band formation in all five samples. When mRNA rather than total RNA was used, nonspecific bands were formed in three of the five samples. Immunobead RT-PCR allowed the detection of 10(1) tumor cells in 1 ml of peripheral blood. The regular RT-PCR analysis had a detection limit of 10(2) tumor cells in 1 ml of peripheral blood. CONCLUSION: The immunobead RT-PCR proved to be more sensitive and specific than the regular RT-PCR at least in our model system. PMID- 11501749 TI - Role of gonadotropins in malignant progression of sex cord stromal tumors produced by sequential auto- and isogenic transplantation of ovarian tissue in ovariectomized rats. AB - PURPOSE: In this study the effect of continuous stimulation of gonadotropins on sex cord stromal tumors in the rat was examined. METHODS: Sex cord stromal tumors were induced by transplantation of ovaries under the splenic capsule of ovariectomized rats. Beginning 180 days after transplantation, these tumors were taken out and cut into several pieces, which were then retransplanted (by isotransplantation) under the splenic capsule of 80 either intact or ovariectomized rats. RESULTS: Most of the tumor grafts grew up to a median size of 0.7 cm in ovariectomized rats. However, some of the tumors recovered from recipient rats that were retransplanted with donor tumors differed significantly from the others. Characterized by a high mitotic rate, nuclear atypia, size (up to 3.8 cm) as well as growth in intact animals, these tumors were defined as malignant. They could be kept in culture and always led to the development of metastases after retransplantation into other rats. CONCLUSION: Benign sex cord stromal tumors can show malignant growth after transplantation. This study for the first time demonstrates that gonadotropins are involved in the induction of ovarian malignancies. PMID- 11501750 TI - Over-expression of beta-1,4-galactosyltransferase I, II, and V in human astrocytoma. AB - PURPOSE: beta-1,4-Galactosyltransferase (beta-1,4-GalT) I, II, and V are the enzymes responsible for the biosynthesis of N-acetyllactosamine on N-glycans by transferring UDP-galactose to the terminal N-acetylglucosamine (N-GlcNAc) residues with the formation of a beta-1,4-linkage. Neoplasms undergo various changes in the carbohydrate of their glycoconjugates, indicating the possible changes in glycosyltransferases themselves. METHOD: Therefore, we compared the expression of beta-1,4-GalTs between astrocytoma and normal brain tissues. RESULTS: Our reverse-transcription polymerase chain reaction (RT-PCR) results showed that beta-1,4-GalT I transcript was absent in normal adult brain but detectable in grade II, III, and IV astrocytomas; the level of beta-1,4-GalT II transcript was increased in grade II, III, and IV astrocytomas while only a trace amount was found in normal brain; beta-1,4-GalT V transcript existed in normal brain and increased in the process of astrocytoma progress, with the highest level in grade IV astrocytoma. By Ricinus communis agglutinin-1 (RCA-1) lectin blot assay, we also found the more extensive galactosylated bands in astrocytomas compared with normal brain. A major 61kD protein was galactosylated in astrocytoma but not in normal brain tissues. CONCLUSION: These results indicate that the increase of galactosylation in astrocytomas may be caused by the alterations of gene expression of beta-1,4-GalT I, II, and V and that the malignant degree of astrocytoma is correlated with the expression of beta-1,4 GalT V. PMID- 11501751 TI - Accelerated hyperfractionated radiotherapy with concurrent chemotherapy in locally advanced nasopharyngeal carcinoma: a phase II study. AB - PURPOSE: The incidence of nasopharyngeal carcinoma in Germany is relatively low in comparison with certain regions in south-east Asia. However, standardised therapeutical regimes are required in the treatment of these tumours. METHODS: Between August 1990 and December 1997, 25 patients with stage III and IV nasopharyngeal carcinoma received an accelerated and hyperfractionated radiotherapy with concurrent chemotherapy (5-FU and mitomycin C). The primary tumour and positive lymph nodes received a total dose of 72 Gy over a period of 6 weeks. In the first 3 weeks, irradiation fields were treated five times per week with 2 Gy per fraction. Thereafter, treatment was accelerated, giving two daily fractions of 1.4 Gy. Salvage surgery was offered for residual lymph node disease after radiotherapy. RESULTS: The overall response rate defined as complete and partial response of the primary was 100%. Sixteen of the 25 patients were still alive and were free of any evidence of tumour recurrence or distant metastases at a mean follow-up period of 34 months (range 7-95 months). Six patients received salvage surgery. Only one of these six patients had histologically proven evidence of vital tumour. No severe late complications such as blindness or temporal lobe necrosis were observed. CONCLUSIONS: The presented data are promising and show that the combination of hyperfractionated accelerated radiotherapy and chemotherapy is feasible and effective. PMID- 11501752 TI - Structural alterations of sugar chains in urine fibronectin from bladder cancer patients and its enzymatic mechanism. AB - PURPOSE: To study the structural alterations of asparagine-linked sugar chains (N glycans) on urine fibronectin (Fn) from bladder cancer (BCa) patients and its enzymatic mechanism. METHODS: Eight pairs of urine samples from eight BCa patients pre-operation and 3 months post-operation (which proved to be normal) were collected, and the Fn in the urine samples was purified with an anti-Fn antibody affinity column. Different lectins labeled with horseradish peroxidase (HRP) were used as probes to bind the glycans of purified Fn immobilized on membrane. Enhanced chemiluminescence (ECL) reagent was adopted to estimate the activity of the bound HRP as a measure of the binding affinity of the Fn glycans to lectins, and expressed as luminescent light units (LLU). The enzymatic mechanism of the structural alteration of N-glycans in BCa Fn was studied by determination of GnT activities using the HPLC method and fluorescent-labeled substrate. RESULTS: The mean LLU of BCa Fn was only 18.1% of the normal samples when Con A-HRP were used as probes, while the mean LLU of the BCa group was 3.34 times and 3.26 times higher than normal for the DSA-HRP and WGA-HRP probes, respectively. The individual data of the patients did not overlap between the BCa sample and normal counterparts, indicating that the positive rates were 100%, regardless of which lectin-HRP was used. These results reveal that the antennary number and bisecting GlcNAc structure are increased in the N-glycans of urine Fn from BCa assessed according to the binding specificity of ConA, DSA, and WGA. In addition, the binding affinities of urine Fn with DSA and WGA were correlated to pathological stage, and the affinity of Fn with WGA was also correlated with pathological grade. The results of GnT determination showed that GnT-III, IV, and V in BCa tissues increased by 34.0, 18.1, and 1.6 times, respectively, in normal bladder tissues which were at least 5 cm away from the BCa of the same bladder. These findings were compatible with the structural changes of N-glycans in BCa Fn, since GnT-III and GnT-IV/V are responsible for the synthesis of bisecting GlcNAc and the increase of antennary number in N-glycans, respectively. CONCLUSIONS: (1) The highest elevation of GnT-III and the close relationship between the WGA binding of BCa Fn with the pathological stage and grade of BCa indicate that the increase of bisecting GlcNAc in N-linked glycans contributes more to the malignant behavior of BCa than the increase of GnT-IV, GnT-V, and the antennary number. (2) The correlation of altered activities of bladder GnTs with the abnormal structures of urine Fn in BCa patients indicates that the urine Fn is synthesized in the bladder. (3) The lectin-HRP assay for analyzing the structure of N-glycans in urine Fn may be used as a simple and accurate diagnosis method for BCa in the future. PMID- 11501753 TI - Biological activity of mammalian transcriptional repressors. AB - Research on the regulation of transcription in mammals has focused in recent years mainly on the mechanism of transcriptional activation. However, transcriptional repression mediated by repressor proteins is a common regulatory mechanism in mammals and might play an important role in many biological processes. To understand the molecular mechanism of transcriptional repression, the activity of eight mammalian repressors or repressor domains was investigated using a set of model promoters in combination with two different transcriptional detection methods. The repressors studied were: REST, the thyroid hormone receptors alpha and beta, the zinc finger protein NK10 containing a 'kruppel associated box' (KRAB), repressor domains derived from the proteins Egr-1, Oct2A and Dr1 and the repressor/activator protein YY1. Here we show that the repressor domains of REST, Egr-1, the thyroid hormone receptors alpha< and beta and NK10 were transferable to a heterologous DNA-binding domain and repressed transcription from proximal and distal positions. Moreover, these repressor domains also blocked the activity of a strong viral enhancer in a 'remote position'. Thus, these domains are 'general' transcriptional repressor domains. The 'kruppel-associated box' was the most powerful repressor domain tested. In contrast, the repressor domains derived from Oct2A and Dr1 were inactive when fused to a heterologous DNA-binding domain. The repressor domain of YY1 exhibited transcriptional repression activity only in one of the transcriptional assay systems. The recruitment of histone deacetylases to the proximity of the basal transcriptional apparatus was recently discussed as a mechanism for some mammalian transcriptional repressor proteins. Here we show here that histone deacetylase 2, targeted to the reporter gene via DNA-protein interaction, functions as a transcriptional repressor protein regardless of the location of its binding site within the transcription unit. PMID- 11501754 TI - Cloning and characterisation of chlorophyll synthase from Avena sativa. AB - The chlorophyll synthase gene from oat (Avena sativa) was cloned and expressed in Escherichia coli. The deduced amino acid sequence consists of 378 amino acids including a presequence of 46 amino acids. Deletion mutants show that a core protein comprising amino acid residues 88 to 377 is enzymatically active. The sequence of the mature protein shows 85% identity with the chlorophyll synthase of Arabidopsis thaliana and 62% identity with the chlorophyll synthase of Synechocystis PCC 6803. The gene is constitutively expressed as the same transcript level is found in dark-grown and in light-grown seedlings. The enzyme requires magnesium ions for activity; manganese ions can reconstitute only part of the activity. Diacetyl and N-phenylmaleimide (NPM) inhibit the enzyme activity. Site-directed mutagenesis reveals that, out of the 4 Arg residues present in the active core protein, Arg-91 and Arg-161 are essential for the activity. Five cysteine residues are present in the core protein, of which only Cys-109 is essential for the enzyme activity. Since the wild-type and all other Cys-mutants with the exception of the mutant C304A are inhibited by N phenylmaleimide, we conclude that the inhibitor binds to a non-essential Cys residue to abolish activity. The role of the various Arg and Cys residues is discussed. PMID- 11501755 TI - DNA damage by 3,6-dihydropyrazine-2,5-dipropanoic acid, the cyclic dimerization product of 5-aminolevulinic acid. AB - 5-Aminolevulinic acid (ALA) is a heme precursor that accumulates in lead poisoning and inborn porphyrias. It has been shown to produce reactive oxygen species upon metal-catalyzed aerobic oxidation and to cause oxidative damage to proteins, liposomes, DNA, and subcellular structures. Studies have also shown that ALA may condense to yield the cyclic product 3,6-dihydropyrazine-2,5 dipropanoic acid (DHPY). Here we propose that DHPY could be involved in DNA damage in the presence of high concentrations of ALA. Exposure of plasmid pUC19 DNA to low concentrations of DHPY (2-10 microM) in the presence of 0.1 mM Cu2+ ions causes DNA strand breaks, as demonstrated by agarose gel electrophoresis. It was also shown that in the presence of Cu2+ ions DHPY is able to increase the oxidation of monomeric 2'-deoxyguanosine to form 8-oxo-7,8-dihydro-2' deoxyguanosine as inferred from high performance liquid chromatography measurements using electrochemical detection. Addition of a metal chelator (bathocuproine, 0.5 mM), the DNA compacting polyamines spermidine (1 mM) and spermine (1 mM) or antioxidant enzymes such as superoxide dismutase (10 microg/ml) and catalase (20 pg/ml) protect the DNA against these damages. The data presented here are discussed with respect to the increased frequency of liver cancer in patients with acute intermittent porphyria. PMID- 11501756 TI - Chicken erythrocyte pyrimidine 5'-nucleotidase: purification and characterization of the subclass I enzyme. AB - Nucleotidase activities resembling subclass I and subclass II of human pyrimidine 5'-nucleotidases (P5N) were detected in chicken red blood cells (RBCs). In chicken RBCs from untreated controls, the activity of the subclass II enzyme was about one third of that of subclass I enzyme, whereas that ratio was approximately 5:1 in rat or human RBCs. The subclass I activity in chicken RBCs was increased 5- to 6-fold upon erythropoietic induction by phenylhydrazine administration, but the subclass II activity did not increase under these conditions. The subclass I enzyme was purified to near homogeneity. Its molecular mass was about 35 kDa as estimated by gel filtration and SDS-polyacrylamide gel electrophoresis. Its N-terminal 12 amino acids, PEFQKKTVHIKD, were also determined. The catalytic properties of the subclass I enzyme were very similar to those of the human enzyme with regard to substrate (preferential hydrolysis of CMP, dCMP, UMP), Km values, optimum pH, and metal ion requirements. Antibodies against chicken P5N subclass I were raised in rats. The chicken P5N-I as well as the rat P5N-I proteins could be detected by antibodies in Western blot analyses, but not the P5N-II proteins. These findings indicate that P5N subclass I may have an important function in chicken erythropoiesis. PMID- 11501757 TI - Coenzymes Q9 and Q10, vitamin E and peroxidation in rat synaptic and non-synaptic occipital cerebral cortex mitochondria during ageing. AB - Great attention has been devoted both to ageing phenomena at the mitochondrial level and to the antioxidant status of membrane structures. These kinds of investigations are difficult to perform in the brain because of its heterogeneity. It is known that synaptic heavy mitochondria (HM) may represent an aged mitochondrial population characterized by a partial impairment of their typical mitochondrial function. We arranged a novel system requiring no extraction procedure, very limited handling of the samples and their direct injection into the HPLC apparatus, to carry out, for the first time, a systematic and concomitant determination of vitamin E, Coenzyme Q9 (CoQ9) and Coenzyme Q10 (CoQ10) contents in rat brain mitochondria. The trends found for CoQ9 and CoQ10 levels in synaptic and non-synaptic occipital cerebral cortex mitochondria during rat ageing are consistent with previous data. Hydroperoxides (HP) differed with age and it was confirmed that in the HM fraction the summation of contributions results in an oxidatively jeopardized subpopulation. We found that vitamin E seems to increase with age, at least in non-synaptic free (FM) and synaptic light (LM) mitochondria, while it was inclined to remain substantially constant in HM. PMID- 11501758 TI - Membrane activity of (Cys48Ser) lung surfactant protein B increases with dimerisation. AB - One of the possible functions of lung surfactant protein B (SP-B), an hydrophobic membrane-associated saposin-like protein, is to reduce the alveolar surface tension by promoting insertion of phospholipids into the air/liquid interface of the lung. SP-B is a covalent homodimer; Cys48 of two polypeptides form an intermolecular disulphide bond. In order to test whether dimerisation of SP-B is important for surfactant function, transgenic mice which express (Cys48Ser) human SP-B in a mouse SP-B null background were generated. In previous studies (Cys48Ser)SP-B showed a concentration-dependent in vitro activity, suggesting that it may form non-covalent dimers. Here (Cys48Ser)SP-B isolated from bronchoalveolar lavage of transgenic mice was studied at different concentrations by circular dichroism (CD) spectroscopy, pulsating bubble surfactometry, mass spectrometry and reversed-phase HPLC. The results indicate that (Cys48Ser)SP-B, both in a phospholipid environment and in organic solvents, is largely monomeric and exhibits low activity at concentrations lower than 1 -2 microM, while at higher concentrations it forms non-covalent dimers, which are nearly functionally equivalent to native SP-B in vitro. Furthermore, electrospray mass spectrometry showed that more dimers were found relative to the monomer when the polarity of the solvent was decreased, and when the concentration of SP-B increased. (Cys48Ser)SP-B also eluted earlier than native SP-B in reversed-phase HPLC. Taken together, these results indicate that a polar surface is buried upon dimerisation, thereby promoting formation of interchain ion pairs between Glu51 Arg52' and Glu51'-Arg52. PMID- 11501759 TI - Participation of residue F552 in domain III of the protective antigen in the biological activity of anthrax lethal toxin. AB - The protective antigen (PA) component of anthrax toxin translocates the catalytic moieties lethal factor (LF) and edema factor (EF) into the cytosol. The proteolytically activated 63 kDa form of PA (PA63) has the ability to oligomerize and bind LF/EF. PA has four distinct domains performing specialized functions; whereas the function of domains I, II and IV has been well characterized, domain III has no known role in the biological activity of PA. Here we report the role of amino acid residues lining an exposed hydrophobic patch of domain III in the biological activity of PA. The residues Phe552, Phe554, lIe562, Leu566 and lle574 were individually substituted with alanine and the effect was studied. All mutant PA proteins except Phe552Ala were equally active as wild-type PA in exhibiting a toxic phenotype to J774A.1 cells in the presence of LF. Substitution of Ala for Phe552 reduced the ability of PA to intoxicate cells by more than 250-fold. However, Phe552Ala was equally active in receptor binding and susceptibility to trypsin and chymotrypsin as wild-type PA, the activities that have been shown to be essential for the biological activity of PA. This mutated PA protein had a decreased ability to bind LF, oligomerize on cells and to induce release of 86Rb+ from Chinese hamster ovary cells. These results suggest that the residue Phe552 in PA plays an important role in LF binding and oligomerization. Our study provides a basis for further exploration of the biological significance of domain III of PA. PMID- 11501760 TI - Purification of chrysancorin, a novel antifungal protein with mitogenic activity from garland chrysanthemum seeds. AB - A novel antifungal protein, designated chrysancorin, was isolated from seeds of Chrysanthemum coronarium var. spatiosum with a procedure involving ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue resin, ion exchange chromatography on SP-Sepharose and FPLC-gel filtration on Superdex 75. The N-terminus of chrysancorin displays sequence similarity to the genomic sequence of chromosome 1 from Arabidopsis thaliana BAC T19E23. Chrysancorin exhibits a molecular mass of 13.4 kDa in gel filtration and SDS-polyacrylamide gel electrophoresis. It stimulates the proliferation of mouse splenocytes and inhibits the activity of human immunodeficiency virus-1 reverse transcriptase. The protein possesses antifungal activity against Botrytis cinerea, Mycosphaerella arachidicola and Physalospora piricola, but not against Rhizoctonia solani, Fusarium oxysporum and Coprinus comatus. However, we could not detect antibacterial activity against a variety of bacteria. PMID- 11501761 TI - Legumain forms from plants and animals differ in their specificity. AB - We purified forms of legumain from a plant source (seeds of kidney bean, Phaseolus vulgaris) and a mammal (kidney of pig, Sus scropha) for comparison of their properties. Both forms were found to be stable only under moderately acidic pH conditions, and were maximally active at about pH 6; the plant enzyme was somewhat less stable and had a slightly higher pH optimum. With benzyloxycarbonyl Xaa-Ala-Asn-aminomethylcoumarylamide substrates, the two forms of legumain showed distinctly different specificities for the P3 residue, the plant legumain preferring amino acids with bulky hydrophobic side chains because of lower Km values. Both forms of legumain were highly specific for hydrolysis of asparaginyl bonds in the arylamide substrates and in neurotensin. Aspartyl bonds were hydrolysed about 100-fold more slowly with lower pH optima. Potential substrates containing other amino acids structurally similar to asparagine were not hydrolysed. There were clear differences in specificity of hydrolysis of protein substrates. The plant legumain differed from pig legumain in its action on tetanus toxoid C-fragment, cleaving at Asn97 but not at Asn337, and produced more extensive digestion of phaseolin. The plant form of legumain was much more weakly inhibited by egg-white cystatin than was the mammalian form. PMID- 11501762 TI - Assignment of the complete disulphide bridge pattern in the human recombinant follitropin beta-chain. AB - The chemical assessment of the complete disulphide bridge pattern in the beta chain of human recombinant follicotropin (betaFSH) was accomplished by integrating classical biochemical methodologies with mass spectrometric procedures. A proteolytic strategy consisting of a double digestion of native betaFSH using the broad-specificity protease subtilisin first, followed by trypsin, was employed. The resulting peptide mixture was directly analysed by FAB MS, leading to the assignment of the first three disulphide bridges. The remaining S-S bridges were determined by HPLC fractionation of the proteolytic digest followed by ESMS analysis of the individual fractions. The pattern of cysteine couplings in betaFSH was determined as: Cys3-Cys5l, Cys17-Cys66, Cys20 Cys104, Cys28-Cys82, Cys32-Cys84 and Cys87-Cys94, confirming the arrangement inferred from the crystal structure of the homologous betaCG. A subset of the S-S bridge pattern comprising Cys3-Cys51, Cys28-Cys82 and Cys32-Cys84 constitutes a cysteine knot motif similar to that found in the growth factor superfamily. PMID- 11501763 TI - The small Gtpase ras is involved in growth factor-regulated expression of the alpha1 integrin subunit in PC12 cells. AB - PC12 cells interact with several growth factors (e. g. EGF, FGF, and NGF) via specific tyrosine receptor kinases, resulting in cell proliferation or neuronal differentiation. The small GTPase Ras is known to be involved in downstream signaling of these growth factor receptors. Furthermore, cell-matrix interactions mediated by integrins, as well as integrin-induced signaling, are also involved in growth factor-stimulated signal transduction in PC12 cells. In this study we determined the expression of the alpha1 integrin subunit in response to EGF and NGF in PC12 wild-type (wt) cells, and in PC12 cells overexpressing an inactive H Ras protein (RasN17). In PC12 wt cells, alpha1 integrin expression is upregulated by EGF and NGF. Cell surface expression of alpha1beta1integrin is also enhanced in growth factor-treated cells. This upregulation leads to increased alpha1beta1 specific adhesion to collagen. In cells expressing the dominant-negative RasN17 variant, alpha1 integrin expression and alpha1beta1-specific adhesion remain unchanged in response to both growth factors. PMID- 11501764 TI - The mouse gap junction gene connexin29 is highly expressed in sciatic nerve and regulated during brain development. AB - A novel mouse gap junction gene, coding for a presumptive protein of 258 amino acids (molecular mass: 28 981 Da), has been designated connexin29. This single copy gene was mapped to distal mouse chromosome 5 and shows 75% sequence identity to a human connexin30.2 sequence in the database. Connexin29 mRNA (4.4 kb) is highly expressed in mouse sciatic nerve and less abundant in spinal cord as well as in adult brain, where it increased 12-fold between day 7 and 14 post partum. Our expression data suggest that the new connexin gene is active in myelin forming glial cells. PMID- 11501765 TI - A selenocysteine-containing peroxiredoxin from the strictly anaerobic organism Eubacterium acidaminophilum. AB - A strongly 75Se-labeled 22 kDa protein detected previously showed in its N terminal sequence the highest similarity to the family of thiol-dependent peroxidases, now called peroxiredoxins. The respective gene prxU was cloned and analyzed. prxU encodes a protein of 203 amino acids (22,470 Da) and contains an in-frame UGA codon (selenocysteine) at the position of the so far strictly conserved and catalytically active Cys47. The second conserved cysteine present in 2-Cys peroxiredoxins was replaced by alanine. Heterologous expression of the Eubacterium acid-aminophilum PrxU as a recombinant selenoprotein in Escherichia coli was not possible. A cysteine-encoding mutant gene, prxU47C, containing UGC instead of UGA was strongly expressed. This recombinant PrxU47C mutant protein was purified to homogeneity by its affinity tag, but was not active as a thiol dependent peroxidase. The identification of prxU reveals that the limited class of natural selenoproteins may in certain organisms also include isoenzymes of peroxiredoxins, previously only known as non-selenoproteins containing catalytic cysteine residues. PMID- 11501766 TI - Characterization of C-terminally truncated human tissue inhibitor of metalloproteinases-4 expressed in Pichia pastoris. AB - The tight regulation of extracellular matrix remodeling and degradation is of great importance in physiological processes like development and morphogenesis, as well as in pathological situations like tumor invasion and metastasis. Tissue inhibitors of metalloproteinases (TIMPs) are the naturally occuring inhibitors of matrix metalloproteinases, which are involved in matrix turnover. In this report we describe the cloning of human TIMP-4 from a human adenocarcinoma and an osteosarcoma cell line and the expression of the inhibitory domain in the methylotrophic yeast Pichia pastoris. The inhibition of MMP-8, -9, -12, -13 and 14 by the N-terminal domain of TIMP-4 was analysed. Using a fluorescent MCA peptide, Ki values for each subclass of MMPs were determined. With dissociation constants in the nanomolar range, TIMP-4 seems to be a good inhibitor for all classes of MMPs without remarkable preference for special MMPs. PMID- 11501767 TI - Gynaecologic oncology: looking back and moving ahead. PMID- 11501768 TI - Ovarian tumor development: insights from ovarian embryogenesis. AB - The study of ovarian embryogenesis can provide important clues about the etiology and development of the different subtypes of ovarian neoplasms. The coelomic epithelium, also called germinal epithelium, was once thought to represent the site of origin of most cellular elements present in the adult ovary. However, recent observations at the morphological, functional, and molecular biological levels strongly suggest that this epithelium plays little or no role in ovarian development. The same observations provide strong support for an important role of the components of the fetal excretory system. These conclusions weaken the hypothesis that the coelomic epithelium is the site of origin of ovarian epithelial tumors. Knowledge of the origin and maturation of germ cells can shed light on several clinico-pathological characteristics of germ cells tumors, including their occasional extra-gonadal origin and differences in the biological behavior of ovarian versus testicular lesions. Knowledge of the mechanisms of regulation of mitotic and meiotic activity during ovarian germ cell maturation can provide insights into the molecular genetic determinants of germ cell neoplasms. The elucidation of molecular pathways actively involved in controlling gonadal differentiation may shed further light into our understanding of the relationship between aberrant differentiation and predisposition to gonadal cancers. PMID- 11501769 TI - Phase II study of a combination of cyclophosphamide, adriamycin and cisplatin in advanced fallopian tube carcinoma. An EORTC gynecological cancer group study. European Organization for Research and Treatment of Cancer. AB - OBJECTIVE: To investigate the clinical activity and toxicity of a combination chemotherapy consisting of cyclophosphamide (C), adriamycin (A) and cisplatin (P) for patients with primary adenocarcinoma of the Fallopian tube having FIGO stage III-IV disease. METHODS: The CAP-regimen consisted of cyclophosphamide 600 mg/m2, adriamycin 45 mg/m2, and cisplatin 50 mg/m2 administered intravenously on day one every 28 days. RESULTS: Twenty-four eligible patients with histologically confirmed Fallopian tube adenocarcinoma were entered in the trial. Fourteen patients had FIGO stage III, and ten had stage IV disease. The median number of CAP cycles was six. Ten patients had a complete and six had a partial response (response rate: 67%, 95% confidence limits: 45-84%). WHO grade III-IV side effects included haematological toxicity, nausea/vomiting and alopecia. Furthermore, mild signs of cisplatin-related peripheral neurotoxicity were observed. At a median follow-up of 40 months, nine patients were alive and 15 had died due to malignant disease. The median time to progression was 13 months for all patients. The median overall survival was 24 months and the 1-, 3- and 5-year survival and their 95% confidence limits were 73% (54-92%), 25% (4-46%) and 19% (0-38%), respectively. CONCLUSION: The present data confirm the therapeutic activity of the CAP-regimen in primary Fallopian tube adenocarcinoma. The response rate is moderate and the toxicity profile is acceptable. PMID- 11501770 TI - Uterine sarcomas in South African black women: a clinicopathologic study with ethnic considerations. AB - BACKGROUND: There is considerable evidence for a higher incidence of uterine sarcomas in blacks when compared to whites. However, whether this higher incidence is related to differences in clinicopathologic presentation is not known. PATIENTS AND METHODS: We reviewed slides and clinical charts of 81 patients with a primary diagnosis of uterine sarcoma referred between 1991 and 1999 to Kalafong Academic and Pretoria Academic Hospital. After review, 49 cases remained for study. RESULTS: Uterine sarcomas were distributed between leiomyosarcoma (LMS) (39%), carcinosarcoma (CS) (49%) and endometrial stromal sarcoma (ESS) (12%). LMS and ESS tend to present at an earlier age when compared to CS (respectively p < 0.008 and 0.02). Of women with LMS more women are premenopausal when compared to CS (p < 0.009). Lower abdominal pain is more common in LMS (p < 0.009), whereas bleeding is more common in women suffering from CS (p < 0.01). Lymphovascular space involvement and cervical involvement are more common in CS when compared to LMS. In CS, the carcinoma component has most of the metastatic potential. CONCLUSION: Among black South African women different clinicopathologic features for uterine LMS, CS and ESS are observed. We also present genetic and/or hormonal factors possibly contributing to the pathophysiology of uterine sarcomas in blacks. PMID- 11501771 TI - MR imaging underestimates stromal invasion in patients with adenocarcinoma of the uterine cervix. AB - PURPOSE OF INVESTIGATION: To evaluate magnetic resonance (MR) imaging in assessing the depth of stromal invasion in patients with adenocarcinoma of the uterine cervix. METHODS: Twenty-three women with adenocarcinoma of the cervix underwent T2-weighted and dynamic MR imaging before surgical evaluation. The images were evaluated for the depth of stromal invasion and compared with histological results. RESULTS: Twelve of 23 patients (52%) were correctly diagnosed in agreement with the histological findings. However, the depth of stromal invasion was underestimated in ten patients (44%). In four of nine patients who exhibited scattered type lesions, no lesions were detected by MR imaging, although deep stromal invasion (more than 1/3) was recognized histologically. In contrast, all 11 patients showed cancer lesions in solid type adenocarcinomas with deep stromal invasion. CONCLUSION: MR imaging detected less stromal invasive lesions in adenocarcinoma of the cervix than surgical specimens, and some scattered type adenocarcinomas could not be visualized. PMID- 11501772 TI - Human papillomavirus infection in the female population of Antwerp, Belgium: prevalence in healthy women, women with premalignant lesions and cervical cancer. AB - Worldwide there is a strong relation between the presence of human papillomavirus (HPV) and the development of cervical cancer. This study investigated the prevalence and genotype of HPV in women with normal smears, women with premalignant lesions and women with cervical cancer in Antwerp, Belgium. Type specific polymerase chain reaction (PCR) for HPV types 16 and 18 and general primer PCR (GP5+/6+) was performed on DNA extracted from paraffin-embedded tissue from women with lesions or fresh material from controls. HPV was detected in 11% of controls, 61% of women with atypia, 77% of women with CIN lesions and 88% of women with cervical carcinoma (chi2 trend, 273, p<0.001). The odds ratio for high risk HPV types was 9.3 for atypia (95%CI. 4.3-19.8), 33.6 for CIN lesions (95%CI, 19.3-58.6) and 78.8 for cervical cancer (95%CI, 39.2-158.3). In total, 19 different HPV genotypes were detected, including five low risk HPV types. Seven of the 14 high-risk HPV types were detected in cervical cancer patients. Based on our study it is suggested that a prophylactic vaccine based on a cocktail of a limited number of high-risk HPV types should be considered in order to protect most women from developing cervical cancer. PMID- 11501773 TI - Assessment of myometrial and cervical invasion of endometrial cancer by transvaginal sonography. AB - INTRODUCTION: The aim of this study was to determine the efficiency of transvaginal ultrasonography in the assessment of myometrial invasion and cervical involvement (preoperative staging) of endometrial cancer. MATERIALS AND METHODS: Transvaginal ultrasonography was performed on 52 women to classify endometrial cancer with respect to myometrial invasion and cervical involvement according to the International Federation of Gynaecologists and Obstetricians recommendations for surgical staging of endometrial cancer. Endometrial cancer was diagnosed on the basis of dilatation and curettage and the degree of invasion was evaluated preoperatively by transvaginal ultrasonography. Ultrasonographic findings were compared to the surgical staging and histopathology of the surgical specimen. RESULTS: Myometrial invasion evaluated by transvaginal sonography was accurate in 46 of 52 cases (accuracy 88%, sensitivity 86%, specificity 90%, positive predictive value 92%, negative predictive value 83%). Tumor extension to the cervix was properly diagnosed in seven of ten women in which it was present. CONCLUSION: Transvaginal ultrasonography is a reliable method for assessing myometrial invasion and cervical involvement. This non-invasive method should be included as an important tool in the establishment of individualized treatment programs for women with endometrial cancer. PMID- 11501774 TI - Adenoid cystic carcinoma of Bartholin's gland: a case report. AB - OBJECTIVE: A case of adenoid cystic carcinoma (ACC) of the Bartholin's gland in a 34-year-old woman with unusual presentation and early recurrence is reported. METHODS: Clinical and histologie features were recorded. Immunohistochemical stains and cell-cycle analysis by flow cytometry technique on paraffin-embedded tumor tissue were performed. RESULTS: The tumor presented as a painful nodule in the episiotomy scar three months after delivery. Initial treatment included only wide local excision. Six months later local recurrence occurred despite clear surgical margins. Histologically a predominant "classic" cribriform growth pattern was identified. Immunoreactivity in tumor cells supported dual epithelial myoepithelial differentiation. Estrogen and progesterone receptors were negative. The DNA histogram revealed a diploid stemline and a low S-phase fraction. CONCLUSION: ACC of the Bartholin's gland is a rare malignant tumor with great propensity for local recurrence. The optimal therapeutic approach has not been established due to the lack of well-defined prognostic parameters. PMID- 11501775 TI - Reduction of visible bone metastases by clodronate therapy in breast cancer. AB - The current report describes a 57-year-old patient with multiple bone metastases 14 years after her initial treatment for breast cancer. The only therapy the patient received for her osteolytic lesions was oral clodronate (800 mg/daily), as she refused any other kind of treatment. On bone scintigraphy the number of visible bone metastases diminished slowly and after two years only a few minor lesions could be seen. Together with this report the value of oral clodronate as anti-osteolytic therapy in breast cancer patients is discussed. PMID- 11501776 TI - Medroxyprogesterone acetate therapy for patients with adenocarcinoma of the endometrium who wish to preserve the uterus-usefulness and limitations. AB - BACKGROUND: To determine the effectiveness of medroxyprogesterone acetate therapy for women with endometrial adenocarcinoma who wish to preserve their uterus. STUDY DESIGN: Fifteen patients with endometrial carcinoma (12 with grade 1 endometrioid adenocarcinoma. 2 with grade 2 adenocarcinoma and 1 with adenoacanthoma) were treated with high-dose medroxyprogesterone acetate alone as primary therapy and their clinical responses evaluated. RESULTS: Seven of the 12 cases (58%) with grade I adenocarcinoma and one of the two (50%) with grade 2 carcinoma responded initially to medroxyprogesterone acetate. The median length of treatment required for regression was 29 weeks. Three patients who initially responded relapsed. Thirteen patients are alive without evidence of disease as of December 1999 (10 to 146 months, median; 4 years and 11 months) and one is continuing medroxyprogesterone acetate therapy as a final follow-up. One patient was lost to follow-up. Two patients have conceived having three healthy infants. CONCLUSION: Treatment of endometrial carcinoma with high-dose medroxyprogesterone acetate could be an alternative to hysterectomy, although the successful rate is limited. PMID- 11501777 TI - A hospital-based multicentric study results on gestational trophoblastic disease management status in a developing country. AB - OBJECTIVE: To determine the clinical management of gestational trophoblastic disease in Turkey. MATERIAL AND METHODS: An inquiry form was sent to 55 health centers including university hospitals, maternity hospitals with residency programs and maternity hospitals without residency programs in 1997. The inquiry consisted of questions about the type of classification systems in use, distribution of cases according to those classifications, use of prophylactic chemotherapy and its indications, and drug preference for single-agent or combined chemotherapies. RESULTS: The overall response rate to the conducted inquiry was 47.1%. A clinical classification system was identified in 60% of the hospitals in Turkey. Generally, methotrexate was the most used single-agent chemotherapy. With regard to first-line combined chemotherapy, MAC (methotrexate, antinomycin-D, cyclophosphamide) was the preferred combination. EMA-CO (etoposide, methotrexate, actinomycin-D, cyclophosphamide, vincristine) was the most common used second-line chemotherapeutic regimen. CONCLUSION: Due to insufficient data acquisition from all the medical centers and a lack of national population-based studies, it is difficult to draw a conclusion with respect to the interpretation of the data about the management protocols of gestational trophoblastic disease. PMID- 11501778 TI - Long-term results from a phase II study of paclitaxel combined with doxorubicin in recurrent platinum refractory ovarian cancer. AB - BACKGROUND: There is still a need for newer non-cross-resistant agents and combinations to be tried in cases of failure after first line platinum-based therapy. Several agents have demonstrated activity after failure of platinum containing regimens. Response rate in true platinum refractory disease up to 20% but with poor long-term survival, has been reported by single drug paclitaxel. In an effort to improve response rate and survival duration obtainable with single drug paclitaxel, we have combined paclitaxel with doxorubicin for the treatment of patients refractory to cisplatin-cyclophosphamide. PATIENTS AND METHODS: Between October 1994 and November 1996, 23 patients whereof 21 refractory to cisplatin-cyclophosphamide were enrolled for toxicity and survival analysis after receiving the combination doxorubicin 50 mg/m2 and paclitaxel 135 mg/m2 every third week for four courses. Responding patients continued on single drug paclitaxel 175 mg/m2 every third week until unacceptable toxicity or tumor progression occurred. RESULTS: The objective response rate (CR + PR) was 33%, 95% CI (14.6-57). The median duration of response was 8.5 months (range 4.0-62.5+) and the median overall survival was 15.5 months (range 4.0-63.5+). No serious toxicity was registered. CONCLUSION: Doxorubicin combined with paclitaxel could safely be administered using this schedule. This study shows that some patients obtaining CR can be rendered disease-free for a substantial period of time, sometimes five years or more. A median overall survival of 15.5 months with a 5 year survival probability of 15% is impressive. However, although responses can be induced in a significant number of patients, the survival figures remain poor. PMID- 11501779 TI - Diagnostic and prognostic value of serum and peritoneal fluid lactate dehydrogenase in epithelial ovarian cancer. AB - OBJECTIVE: The aim of this study was to investigate the peritoneal fluid and serum lactate dehydrogenase (LDH) levels in patients with ovarian masses. MATERIALS & METHODS: Peritoneal fluid and serum lactate dehydrogenase (LDH) levels were measured in 27 patients with epithelial ovarian carcinoma and 38 with benign ovarian tumors. Serum and peritoneal fluid LDH levels were also compared with the levels of CA-125. RESULTS: Both of the marker levels in ovarian cancer patients were significantly higher than those in patients with benign ovarian tumors. Serous and undifferantiated carcinomas presented higher marker levels than endometrioid and mucinous carcinomas. High grade, advanced stage and positive cytology were associated with higher serum and peritoneal fluid LDH levels; there was an inefficient correlation between them but, when these two markers were used together with CA-125, sensitivity of CA-125 increased to 70%. CONCLUSIONS: In conclusion, serum LDH can be used to discriminate adnexal mass origin and peritoneal fluid LDH may have prognostic value because of the strict relationship with advanced stage, poor histologic type, higher grade and positive abdominal cytology. Peritoneal LDH is found to be a reliable biochemical marker related to prognosis in ovarian carcinoma patients. PMID- 11501780 TI - Replacement of the uterus by malignant mesothelioma of the peritoneum: a case report. AB - Malignant mesothelioma of the peritoneum is a rare tumor that must be distinguished from the more common primary peritoneal serous epithelial neoplasms. We report a case of a 66-year-old female presenting with weight loss, anemia, and a large pelvic mass on ultrasound. At laparotomy a large (9 x 6.5 x 3.5 cm) mass in the anatomical position of the uterus was found in the minor pelvis. The uterus, tubes and ovaries could not be identified. Palpable paraaortic lymph nodes and liver nodules were found. Immunohistochemically the tumor cells were positive for cytokeratin, epithelial membrane antigen and vimentin-CEA, but S-100 protein and Leu-M1 were negative. Remnants of the uterine corpus, fallopian tubes and ovaries could not be identified. The overall features were best regarded as malignant peritoneal mesothelioma. There have been reports of mesotheliomas involving the uterus. However, this is the first reported case of mesothelioma causing total replacement of the uterus. PMID- 11501781 TI - Cervical intraepithelial neoplasia in the young female. Diagnosis and management. AB - The prevalence of HPV and CIN in young women has increased in recent years. During a 5-year period (1996-2000), 78 sexually active young females, aged 15-20 years, were referred to the Colposcopic Unit of the 1st Department of Obstetrics and Gynecology of the University of Athens in the major University-appointed hospital in Greece, because of an abnormal cytology or a suspicious cervical abnormality in the presence of negative cytology. Colposcopic examinations were found to be within normal limits in 12/78 (15.4%) of cases. Cervical pathology was related in 22 cases (28.2%) to HPV infection, 23 (29.5%) cases to CIN 1, 18 (23.1%) cases to CIN II and 3 (3.8%) to CIN III. No relation between oral contraceptive use and cigarette smoking with HPV infection was found. Our findings strongly confirm the necessity of obtaining cervicovaginal smears on all sexually active gynecologic and obstetric teenage patients. PMID- 11501782 TI - Squamous cell carcinoma arising in mature cystic teratoma of the ovary: a case report. PMID- 11501783 TI - Antimetastasis gene expression and numerical chromosomal abnormalities of chromosomes 1 & 17 in serous tumours of the ovary. AB - OBJECTIVE(S): The aim of this study was to examine the expression of the antimetastasis gene nm23 and numerical changes on chromosome 1 and 17 in ovarian tumours. METHODS: In this study 20 serous cystadenocarcinomas, ten borderline and five benign tumours were analysed for expression of the nm23 antimetastasis gene by immunohistochemistry and for numerical chromosomal abnormalities of chromosomes 1 and 17 by interphase cytogenetics. RESULTS: Strong intracytoplasmic immunoreactivity with the antimetastasis gene was observed in late stage carcinomas but not in benign or borderline tumours or in lymph node metastases. Numerical abnormalities were only observed in carcinomas. CONCLUSION(S): These sets of data are consistent with the majority of benign and borderline tumours lacking invasive potential. Odds Ratio (OR) assessment indicates that the presence of numerical aberrations correlates with immunopositivity. PMID- 11501784 TI - Ovarian tumors in young females. A laparoscopic approach. AB - Ovarian tumors are one of the major preoccupations in the everyday practice of gynecology. During the period January 1997 through December 2000, 54 cases of ovarian tumors in young females aged 14-20 years were diagnosed and managed laparoscopically in our institution. Twenty-two cases of mature cystic teratoma, 12 cases of endometriosis, eight cases of serous cystadenoma, five cases of mucinous cystadenoma, three cases of fibroma-thecoma, two cases of serous low malignant tumors and one case of mucinous low-malignant tumor were found. The management of ovarian tumors during this age by laparoscopic techniques represents an efficient and safe procedure. PMID- 11501785 TI - Effect of hydrophobic permeation enhancers on the release and skin permeation kinetics from matrix type transdermal drug delivery system of ketotifen fumarate. AB - Ketotifen fumarate is effective in low doses in the treatment of bronchial asthma particularly of allergic origin. However, it is substantially metabolized in the liver when administered orally. Hence transdermal patches of combination of ethylcellulose/polyvinylpyrrolidone and Eudragits RS 100/RL 100 were prepared and their drug release kinetics and skin permeation profiles were evaluated. However, the skin permeation profiles were found to be low and subtherapeutic. Hence three hydrophobic biocompatible substances, viz, isopropyl myristate, isopropyl palmitate and linoleic acid and also combination of isopropyl myristate and linoleic acid were used as permeation enhancers in the film. It was found that isopropyl myristate and linoleic acid combination and isopropyl myristate alone produced promising results compared to isopropyl palmitate and linoleic acid. PMID- 11501786 TI - Effect of hydrophilic substances on liberation of quinidine from starch - alginate sphere. AB - The effect of selected hydrophilic agents on the quinidine loading and its release from potato starch alginate spheres prepared by instilling viscous suspensions into a desolvation liquid (saturated calcium chloride: paraffin liquid: heptan 1:1:1 v/v/v) was studied. The loading of quinidine in spheres was 59.77%-46.30% and it depended on the presence of auxiliary hydrophillic agents such as polyethylene glycol 2000 (59.77%), polyoxyethylene - 20 - sorbitan monostearate (49.55%), glyceryl monostearate (49.02%), sorbitan monooleate (46.30%). The drug release from spheres was diffusion controlled in accordance with the Higuchi model. It was concluded that the release rate constants depended on the drug - carrier - hydrophilic agent composition and dissolution media. PMID- 11501787 TI - Effect of pH and temperature on the interaction of methotrimeprazine with cyclodextrins. PMID- 11501788 TI - Solubility and dissolution rate of ibuprofen in ionic and non-ionic micellar systems. AB - Equilibrium solubility, the profile and the rate of ibuprofen dissolution in solutions of sodium dodecyl sulfate, Tween 60 and Brij 35 were studied. It was found that the solubility of ibuprofen was enhanced by the formed micellar system. The size and shape of formed micelles depended on the used particular surfactant. The mechanism of ibuprofen dissolution in micellar system was discussed. PMID- 11501789 TI - The development and validation of the HLPC method for morphine content determination in poppy straw. AB - The HPLC method for morphine content determination in poppy straw has heen developed and validated. The method validation involved reproducibility and the following parameters of accuracy: selectivity and specificity, linearity and limits of detection and determination, interferences and recovery. The method was used to determine the morphine content in plant resources classified by the Law on "Neutralization of drug abuse" as a intoxicating agent from group I-N, in the content range of 0.02 to 0.27%. The determination was established for 80 samples of plant resources from the whole country of Poland. PMID- 11501790 TI - Flavonoid compounds in the flowers of Kitaibelia vitifolia Willd. (Malvaceae). AB - The following compounds from the flowers of Kitaibelia vitifolia were isolated and identified: kaempferol 3-O-(6"-p-coumaroyl)-beta-glucoside (trans tiliroside); quercetin and kaempferol - 3-O-beta-xylopyranosyl (1-->2)-beta glucopyranoside (3-O-sambubioside): quercetin and kaempferol - 3-O-sambubioside-7 O-glucoside; apigenin, 7-O-sambubioside, luteolin and chrysoerioL, 7-O xylosylglucosides also apigenin 7-O-alpha-rhamnopyranosyl (1-->2)-beta glucopyranoside. PMID- 11501791 TI - Radermachera xylocarpa: the highly efficient source of lapachol and synthesis of its derivatives. AB - Isolation and characterization of the major chemical constituents from the stem bark of Radermachera xylocarpa. Chloroform extract of the powdered stem bark contains mainly two components: one is lapachol (91.2%) and second is alpha lapachone (5.0%), which are reported herewith first time from this plant, in good quantity and purity. PMID- 11501792 TI - Effect of epsilon-aminocaproylaminoacids on the activity of proteolytic enzymes. AB - Effect of three epsilon-aminocaproylaminoacids with significant antifibrinolytic activity on chymotrypsin, trypsin, cathepsin B, cathepsin C and cathepsin D activities was examined. Slight inhibition of trypsin and chymotrypsin activity was observed only at high concentrations of these compounds. All tested dipeptides did not influence activities of cathepsin B, cathepsin C and cathepsin D. PMID- 11501793 TI - Lack of effect of physical exercise on pharmacokinetics of acetaminophen tablets in healthy subjects. AB - The study was designed to evaluate the effect of physical exercise on the pharmacokinetics of acetaminophen. Ten healthy volunteers were drawn to the investigation. Each person was subjected to the pharmacokinetic study twice, i.e. at rest and on an exercise day. On the exercise day the subjects were performing 3 hours long physical exercise which consisted of a walk on a treadmill at 3 mph for 20 minutes of each half an hour. Acetaminophen was given orally at a single dose of 1.0 g prior to the exercise. Blood for pharmacokinetic assay was sampled within 24 hours following the drug administration. Acetaminophen concentrations were determined by the FPIA method. The total concentrations versus time curve were described by the one-compartment open model for extravascular administration. It was revealed, that the standard submaximal physical exercise does not affect the pharmacokinetics of acetaminophen statistically significantly. PMID- 11501794 TI - Conformational analysis and pharmacophore design for selected 1-(2 pyrimidinyl)piperazine derivatives with sedative-hypnotic activity. AB - The conformational of selected 1-(2-pyrimidinyl)piperazine derivatives with high sedative-hypnotic activity was analysed and the model bioactive conformations were suggested. Subsequently, the pharmacophores of analysed compounds were designed. It was suggested that the pharmacophore of bioactive derivatives should be composed of 11 features that characterise the binding model of pyrimidinylpiperazine ligands to the binding site at the hypothetic receptor. This 11 feature pharmacophore was compared to three other pharmacophores designed for the selected anxiolytics (benzodiazepines and buspirone analogues) and the sedative-hypnotic agents (benzodiazepines and barbiturates). Several substantial differences between the pharmacophores were found: the number of pharmacophoric features and their distribution in 3-D space were unique for selected groups of compounds that exhibit sedative-hypnotic or anxiolytic activity. PMID- 11501795 TI - Application of ammonium peroxidisulfate and metavanadate for spectrophotometric determination of prothipendyl hydrochloride. AB - Prothipendyl hydrochloride reacts with (NH4)2S2O8 and NH4VO3 forming the coloured oxidation products which exhibit maximum absorbance at lambda = 372 nm and 374 nm, respectively. The optimum conditions of the reaction have been established. It was found that Beer's law is obeyed in the concentration range 3-95 microg/ml and 3-90 microg/ml in PTP-(NH4)2S2O8 and PTP-NH4VO3 system, respectively. PMID- 11501796 TI - Elaboration of HPLC method for biotin determination in multiple vitamin drugs and comparison with microbiological method. AB - We were searching for optimal analytical conditions for the biotin determination by the HPLC method with detection at the wavelength 200 nm in multiple vitamin drugs. Statistical parameters of the HPLC method and the microbiological method of the biotin determination, were compared. PMID- 11501797 TI - Determination of moclobemide, paroxetine, and fluvoxamine in tablets by HPLC. AB - A simple, accurate high performance liquid chromatography procedure is presented in order to determine moclobemide in Aurorix-tablets, paroxetine in Seroxat tablets and fluvoxamine in Fevarin tablets. These drugs were chromatographed on a Nova-Pak C18, (dp = 4 microm), 150x3.9 mm i.d. column using methanol/phosphate butter adjusted to pH 2.65 with phosphoric acid (7:3, v/v) as the mobile phase to determining of the moclobemide and methanol/tetrahydrofuran/phosphate buffer at pH 2.65 (53:5:4, v/v) for determining of paroxetine and fluvoxamine. The detection was carried at 235 nm. The method was tested for linearity over the range 5-50 microg/ml. The relative standard deviation for moclobemide is 1.16, but for paroxetine and fluvoxamine is less than 1% (0.25 and 0.46 respectively). PMID- 11501798 TI - Dome (modified Chiari) pelvic osteotomy: 10- to 18-year followup study. AB - Ninety-six dome (modified Chiari) pelvic osteotomies in 87 patients with pain and disability because of osteoarthrosis secondary to hip dysplasia were reviewed. The mean age of the patients at the time of surgery was 29 years (range, 16-55 years). The mean followup was 13 years (range, 10-18 years). Forty-one hips were classified into a hip dysplasia stage, 32 hips into an early stage of osteoarthrosis, and 23 hips into an advanced stage of osteoarthrosis according to the radiographic grading of the Japanese Orthopaedic Association. The average preoperative Merle d'Aubigne and Postel hip score was 13.8, and the average score at final followup was 16.6. Excellent or good results with a score greater than 14 were achieved in 96% of the hips at final followup. Radiographically, signs of progression of osteoarthrosis were not seen in 87% of the hips. Osteoarthrosis progressed during the postoperative course in 13% of the hips. Four patients (four hips) eventually had a total hip arthroplasty at 13.8 years (range, 12.5 15.3 years) after surgery. The survival rate of dome pelvic osteotomy, using clinical failure as an end point, was 82% at 15 years. Using radiologic failure as an end point, 61 hips in 54 patients (survival rate, 78%) survived at 15 years. Using hip replacement as an end point, 92 hips in 83 patients (survival rate, 82%) survived at 15 years. Dome pelvic osteotomy is an excellent and effective operation for pain relief and functional maintenance of the hip. PMID- 11501799 TI - Debris from failed ceramic-on-ceramic and ceramic-on-polyethylene hip prostheses. AB - To compare the properties of wear debris between ceramic-on-ceramic and ceramic on-polyethylene total hip prostheses, particles were isolated and characterized from tissue biopsies obtained at revision arthroplasty or autopsy from two similar uncemented modular hip systems. Group A hips (11 patients; mean, 31 months in vivo) had titanium shells with alumina inserts, alumina femoral heads, and titanium alloy stems. Group B hips (seven patients; mean, 42 months) were the same as Group A but with polyethylene acetabular inserts. Particles were characterized using an electrical resistance particle analyzer, scanning electron microscope, and energy dispersive xray spectroscope. Most of the particles in Group A were ceramic, whereas most of the particles in Group B were polyethylene. Metal particles from the femoral stem and the acetabular shell also were present. If one Group A hip with impingement is excluded, the rate of particle production is significantly lower in the ceramic-on-ceramic group than in the ceramic-on polyethylene group. With the number of samples available, no significant difference in average size could be detected among the different types of particles or among the groups. PMID- 11501800 TI - Results of hip revision using the Exeter stem, impacted allograft bone, and cement. AB - The Exeter stem and impacted, morselized allograft bone and cement were used in the revisions of 18 consecutive femoral components (17 patients). The primary arthroplasty had been done because of osteoarthritis. All of the femoral components were revised for the first time because of aseptic loosening. The migration pattern of the Exeter stem after revision was studied using roentgen stereophotogrammetric analysis. At 2 years after surgery, all 18 femoral stems had migrated in the distal direction (average, 2.5 mm). In addition, seven of the stems had migrated in the medial direction (average, 1.3 mm), and two stems had migrated in the lateral direction (0.5 mm and 1 mm, respectively). Sixteen of the femoral stems also had migrated in the posterior direction (average, 2.9 mm), but none migrated in the anterior direction. The migration rate decreased gradually with time during the followup. Six femoral stems continued to migrate between 1.5 and 2 years after surgery. In patients with major femoral bone deficiency at the time of hip revision, the use of impacted morselized allograft bone and cement yielded an initial fixation similar to that obtained in conventionally cemented revisions. Pain had improved in all patients at the 2-year followup. PMID- 11501801 TI - Natural progress of a bone scan after cementless hydroxyapatite-coated total hip arthroplasty. AB - To establish the natural progress pattern of postoperative bone uptake, a periprosthetic quantitative technetium-99m methylene diphosphonate scintiscan was done on 80 asymptomatic hips (62 patients) with uncomplicated cementless hydroxyapatite-coated total hip arthroplasties and 20 healthy control hips (10 subjects) without previous surgery. The patients were studied in eight groups at scheduled intervals of 1 to 48 months. There were 10 hips in each group. The measurement of bone uptake in the healthy untreated control group indicated that the uptake ratio in the proximal femur was physiologically higher in the metaphyseal area than in the diaphyseal area and the uptake ratio in the acetabulum appeared to be much higher than that of the proximal femur. In the patient group, the uptake ratio around the femoral stem area and the acetabular cup area showed a statistically significant decrease between 1 and 3 months after surgery and changed little after 3 months. Comparing the result of the patient group with that of the healthy untreated control group, the uptake ratio decreased much faster in the hydroxyapatite-coated metaphyseal zone than in the noncoated diaphyseal zone of the femoral stem area. In the acetabular cup area, the uptake ratio decreased fast, as in the hydroxyapatite-coated metaphyseal zone of the femoral stem area. Based on these clinical results, a quantitative bone scan may be a helpful diagnostic procedure for evaluating postoperative progress when used in conjunction with clinical symptoms and radiologic examinations. PMID- 11501802 TI - Knee arthrodesis after infected total knee arthroplasty using the Ilizarov method. AB - Numerous treatments have been proposed for infected total knee arthroplasty. In selected patients, a knee arthrodesis is a well-recognized salvage procedure. However, there are no reports in the literature discussing the treatment of infected total knee arthroplasty using the Ilizarov method. The authors reviewed their experience with this technique in infected total knee arthroplasty, presenting six patients (four women, two men) treated between 1992 and 1998. The average age was 56.6 years (range, 23-70 years) and the mean number of previous surgical procedures was seven (range, 4-10 procedures). From the time of frame removal, the patients were followed up for a mean of 34.2 months (range, 12.4 87.5 months). Full weight-bearing was allowed 1 week after surgery if half-pins were used and after a mean of 2.6 months if Kirschner wires were used. Of the five patients who have completed treatment, all have obtained a stable knee arthrodesis after a mean external fixation time of 6.8 months without additional surgical procedures or bracing. All of the patients were satisfied with the treatment. The authors recommend knee arthrodesis by the Ilizarov method for infected total knee arthroplasty, particularly in patients with extensive bone loss, significant limb shortening or axial deformity or both, active infection, or previous failed arthrodesis. PMID- 11501803 TI - Patellofemoral imaging. AB - Imaging of the patella and patellofemoral joint cannot be isolated from imaging of the knee anymore than the function of the patellofemoral joint can be separated from the function of the entire knee. Therefore, the current author will focus on imaging of the patellofemoral joint yet integrate this with the entire knee. Because radiographs and other imaging techniques are adjuncts to the history and physical examination, and specifically an extension of the physical examination, each view or technique must be selected thoughtfully to yield the most useful and accurate information possible. Imaging techniques range from relatively inexpensive to very costly, yet the amount of useful information may not show a positive correlation with the cost. Only by integrating the history, physical examination, and the best imaging techniques can the clinician locate the abnormality, determine its cause, and plan a proper course of treatment. Radiographic imaging of the patellofemoral joint requires attention to detail on the part of the technician and knowledge of these techniques on the part of the clinician who orders and interprets the results. Understanding these concepts will help achieve everyone's goal: high-quality and cost-effective orthopaedic care. PMID- 11501804 TI - Observations of the proximal tibia in total knee arthroplasty. AB - Relatively little has been written concerning the proximal tibia in total knee arthroplasty. Few authors have looked at landmarks and guidelines for tibial tray preparation and tibial tray orientation. The current study showed that a line drawn 1 mm medial to the medial border of the tibial tubercle and going through the midsulcus of the tibial spines (the midsulcus line) provided a reproducible landmark for the tibia, and when a perpendicular cut was made relative to this line, 46 of 50 knees were cut in appropriate alignment. In addition, it has been said that the tibial tray should be rotated externally to approximately the medial 1/3 of the tubercle to maximize function. The current study showed that when the tibia is allowed to float in a functional position relative to the femoral implant, the tibial external rotation was only 2 mm lateral from the medial age of the tibial tubercle; this is far less than the medial 1/3 of the tubercle and close to the starting point of the midsulcus line. PMID- 11501805 TI - Surgical treatment of chronic lower extremity neuropathic pain. AB - The current authors retrospectively reviewed 147 lower extremity peripheral nerve procedures in 114 patients (average age, 42 years) with chronic lower extremity neuropathic pain to determine whether surgical treatment based on an empirically derived algorithm could reduce pain and improve function. This algorithm assigns crush, stretch, and chronic transection injuries to treatment with transection and containment. Peripheral nerve stimulation was used in conjunction with transection and containment for patients with more chronic presentations for whom previous transections had been unsuccessful. Patients with adhesive neuralgia underwent revision neurolysis with vein wrapping. Patients with repetitive nerve trauma (overuse) underwent primary or revision neurolysis. Duration of symptoms averaged 37 months, and mechanisms of nerve injury included chronic transection, crush, adhesive neuralgia, stretch, repetitive trauma, and idiopathic etiology. Time to followup averaged 38 months. Pain and dysfunction were ranked from 0 points (no pain or dysfunction) to 10 points (pain prompting request for amputation or functional deficit warranting wheelchair use); preoperative and followup work status were documented. Average pain and dysfunction scores improved: 8.8 to 5.6 points and 7.6 to 5.0 points, respectively. Of the 114 patients, 52 (46%) patients improved their work status, including 35 of 87 (40%) involved in workers' compensation. There were no statistically significant differences in outcome based on mechanism of nerve injury or type of procedure. The consistent average improvement suggests this algorithm assigns the appropriate procedure to a given mechanism of injury. PMID- 11501806 TI - Pathologic fractures after surgery and radiation for soft tissue tumors. AB - Twenty pathologic fractures occurred in 285 patients with soft tissue tumors that were treated with radiation therapy and surgery. Twelve of these fractures occurred in 11 patients from a subpopulation of 163 patients with 168 soft tissue tumors of the thigh (155 soft tissue sarcomas and 13 aggressive fibromatosis). The fractures occurred at a mean of 40.5 months after treatment and were not associated with significant trauma. Risk factors associated with the development of fracture included tumor location within the anterior compartment of the thigh, extensive surgical periosteal stripping, and a marginal or intralesional margin of resection. The dose, timing, and fractionation of radiation therapy were not related to the risk of fracture. A high rate of complications was seen with this series, including fracture nonunion (45%) and deep infection (20%). Prophylactic intramedullary fixation of the femur should be considered for patients undergoing resection of large tumors in the anterior compartment of the thigh requiring extensive periosteal stripping and adjuvant radiation therapy. PMID- 11501807 TI - Functional evaluation in distal femoral endoprosthetic replacement for bone sarcoma. AB - A multicenter study of successfully treated patients (mean age, 36.7 years) with a minimum 1-year followup (average, 35.4 months) after distal femoral endoprosthetic replacement for bone sarcoma was done using the 1987 and 1993 versions of the Musculoskeletal Tumor Society, the Short Form-36, and the Toronto Extremity Salvage Score functional evaluation criteria. Fifty-six patients (28 women and 28 men) fulfilled the criteria. Thirty-one Kotz prostheses (fixed hinge, uncemented) and 25 Modular Replacement System Prostheses (rotating hinge, cemented) were used. Thirty-five patients walked without aids, 19 used a cane, and two used crutches or a walker. The Musculoskeletal Tumor Society 1987 mean score was 28.1. The Musculoskeletal Tumor Society 1993 mean score was 80.4. The Toronto Extremity Salvage Score mean was 81.6. The Short Form-36 Physical Component Score had a mean of 43.2 and Mental Component Score mean of 54.2. The two groups of implants were comparable, except for the length of bone resection. Multivariate regression analysis revealed that patient age, existence of a pathologic fracture, and type of prosthesis all significantly accounted for differences in functional outcome as measured by the Musculoskeletal Tumor Society 1993, the Toronto Extremity Salvage Score, and the Short Form-36 Physical Component Score scales. Although both implants provided satisfactory function, the Musculoskeletal Tumor Society 1993 and the Toronto Extremity Salvage Score results were significantly better with the Modular Replacement System prosthesis. The effect of possible differences among surgeons or institutions was not addressed. PMID- 11501808 TI - Parosteal lipoma of the distal phalanx: a case report and review of the literature. AB - A rare case of parosteal lipoma of the distal phalanx in a small finger of a 68 year-old woman is reported. Radiographs revealed an osseous projection and cortical irregularity in the distal phalanx of the right small finger. Magnetic resonance imaging showed a hyperintense soft tissue mass adjacent to the phalanx on T1-weighted and T2-weighted images. Histologically, the lesion consisted of mature fat cells. Marginal excision of the lesion was curative. PMID- 11501809 TI - Calcific myonecrosis of the leg: a case report and review of the literature. AB - Calcific myonecrosis is a rare complication of the leg after trauma. It usually develops as a late sequelae of compartment syndrome. Only approximately 21 cases have been reported in the English literature. The reported cases mostly have involved the anterior compartment of the leg. The current authors report the case of a 49-year-old man with a huge mass that involved the anterior and posterior compartments of the leg 30 years after initial trauma. Soft tissue sarcoma was suspected initially. A secondary infection developed after open incisional biopsy. After extensive debridement and closure of the wound over antibiotic impregnated cement beads and a suction drain followed by compressive dressings, there has been no recurrence of the lesion or infection. PMID- 11501810 TI - Type IV tibial tubercle fracture revisited: a case report. AB - Tibial tubercle avulsions and proximal tibial physeal fractures are uncommon. An even more rare case was seen when an adolescent athlete presented with a combined Type III tibial tubercle avulsion and a Salter-Harris Type IV proximal tibial physeal fracture. There was no previous report in the literature of the combination of these two uncommon fracture patterns, both of which are associated with serious potential complications and long-term sequelae. The two fracture patterns must be recognized individually, with treatment directed appropriately. PMID- 11501811 TI - Treating forearm fractures using an internal fixator: a prospective study. AB - Some major complications of internal fixation with plates, such as infections and disturbance of healing, have been shown to be related to necrosis of bone and to the soft tissues immediately deep to the plate. This is attributable to plate contact. To deal with this phenomenon, an internal fixator, the Point Contact Fixator, was developed according to a new concept. The Point Contact Fixator resembles a plate but functions like a fixator, that is, the fracture is stabilized using a splint fixed to the bone by monocortical, angularly locked screws that are designed not to exert pressure between the splint and the bone, thereby minimizing implant-to-bone contact. Vascular damage to the osseous blood supply consequently is avoided. The new internal fixator is the first of a new family of implants in addition to nails, plates, and external fixators. To study the potential of the Point Contact Fixator in a prospective study, 79 forearm fractures in 55 patients were treated in a consecutive series by one surgeon using the same technique throughout. Followup to union is reported for 100% of the patients. Handling the fixator was simple; healing was uneventful; and the rate of complication was low. PMID- 11501812 TI - Cerebral fat embolism after a nondisplaced tibial fracture: case report. AB - Fat embolism syndrome has been observed after traumatic or nontraumatic events. In traumatic cases, fat embolism syndrome is known to occur in patients with a fracture of a long bone. The case of a patient with a cerebral fat embolism associated with a nondisplaced fracture of the tibial shaft is reported. PMID- 11501813 TI - Percutaneous implantation of iliopsoas for functional neuromuscular stimulation. AB - Hip flexion is required for walking and stair climbing. Percutaneous electrical stimulation of the iliopsoas muscle is a potentially useful and reliable method of providing hip flexion in individuals who are paralyzed. In this study, groin, lateral abdominal, and paraspinal approaches of percutaneous electrode implantation for electrical stimulation of the iliopsoas muscle are described. The paraspinal approach using stimulation of the second and third lumbar roots gave the best hip flexion response; however, it often was accompanied by unwanted stimulation of the hip adductor and abdominal muscles. Wire breakage and electrode movement were the most common causes for failure of maintaining hip flexion. The paraspinal approach, using double helix electrodes, provided an average of 110 weeks of functional hip flexion sufficient for walking. It is feasible to implant electrodes in the iliopsoas muscle. An open technique for permanent implantation of intramuscular electrodes is being developed to selectively stimulate the iliopsoas, which will extend the range and duration of hip flexion that will allow stair climbing in individuals who are paraplegic. PMID- 11501814 TI - Effect of femoral stiffness on bone remodeling after uncemented arthroplasty. AB - The current study examined the relationships among femoral stiffness, implant stiffness, and bone remodeling in 40 femurs retrieved at autopsy from 20 patients with unilateral uncemented hip replacements. The purpose of the study was to determine if the magnitude of periprosthetic bone loss after arthroplasty was correlated with, and could be predicted from, stem and femoral stiffness terms. For analysis, the contralateral normal femur was used as a control to represent the unremodeled condition of the in vivo implanted femur. Bone loss attributable to remodeling was quantified by video-densitometric analysis. Stiffness terms were calculated as the product of the elastic modulus and geometric properties digitized from cross-sectional slab radiographs. Femoral stiffness calculations accounted for variations in modulus attributable to patient differences in bone mineral density and geometric properties attributable to differences in the shape of individual femurs. Similarly, calculations of implant stiffness accounted for variations in implant shape. Results showed axial bone stiffness was the variable most strongly correlated with bone loss. Individual stem stiffness terms were not significantly correlated with bone loss. Multiple linear regression analysis, using stem-to-bone stiffness ratios as independent variables, accounted for 46% of the variance in bone loss data. In the regression analysis, the axial stem-to bone stiffness ratio was the strongest correlate with bone loss. Although these results show the influence of mechanical stiffness factors on bone remodeling, other factors (hormonal status, drugs, disease, activity level) could represent the variance in bone loss data not accounted for in the study. PMID- 11501815 TI - Congenital dislocation of the patella. AB - Congenital dislocation of the patella may occur as a persistent lateral dislocation of the patella that presents with a knee flexion contracture and the patella tethered lateral to the femoral condyles or as an intermittent dislocation of the patella. In the latter syndrome, the patella dislocates completely with each flexion and extension cycle of the knee and is best termed obligatory dislocation of the patella, because the patient has no control over the patella dislocating as he or she moves the knee. The first type of congenital dislocation, which is fixed, often is associated with syndromes such as arthrogryposis and should be corrected surgically by lateral release and realignment of the patella. Obligatory dislocation of the patella tends to be an isolated dysplastic anomaly and may be relatively well tolerated. Rebalancing of the patella usually is done at a later age because of less interference with function. The current author describes the natural history of patella femoral dysplasia, detailing the pathologic changes that are present, and recommends surgical techniques for correcting both types of congenital dislocation of the patella. PMID- 11501816 TI - Donor cell fate in tissue engineering for articular cartilage repair. AB - Articular cartilage repair is a clinical challenge because of its limited intrinsic healing potential. Considerable research has focused on tissue engineering and transplantation of viable chondrogenic cells to enhance cartilage regeneration. However, the question remains: do transplanted allogenic cells survive in the repair with time? This study assessed donor cell fate after transplantation of male New Zealand White rabbit perichondrium cell and polylactic acid constructs into osteochondral defects created in the medial femoral condyles of female New Zealand White rabbits. Repair tissue was harvested at 0, 1, 2, 3, 7, and 28 days after implantation and was evaluated for cell viability and total cell number using confocal microscopic analysis. The number of donor cells in each sample was estimated using quantitative polymerase chain reaction targeting a gender-specific gene present on the Y-chromosome, the sex determining region Y gene, and a control deoxyribonucleic acid present in male and female cell deoxyribonucleic acid, the matrix metalloproteinase-1 gene promoter. Average cell viability was found to be 87% or more at all times. Donor cells were present in repair tissue for 28 days after implantation. However, the number of donor cells declined from approximately 1 million at Time 0 to approximately 140,000 at 28 days. This decline in donor cells was accompanied by a significant influx of host cells into the repair tissue. This study shows that the sex-determining region Y gene is a valuable marker for tracking the fate of transplanted allogenic cells in tissue engineering. PMID- 11501817 TI - Evaluation and treatment of dural tears in lumbar spine surgery: a review. AB - Incidental durotomy is a frequent complication of lumbar spinal surgery. The number and complexity of spinal procedures is increasing, leading to a greater prevalence of dural tears; therefore, it is imperative that spine surgeons be familiar with safe and effective closure techniques. Occasionally, a tear may not be recognized during the procedure, so that one must recognize the signs and symptoms of a cerebrospinal fluid leak postoperatively. Several newer treatment concepts show promise. The current study represents an extensive review of the recent literature on the prevalence, mechanism, diagnosis, treatment, and outcomes of dural tears. The authors provide an overview of the problem, an update on current treatment strategies, and describe the senior author's technique of repair, which is easy to do and is effective in stopping additional leakage of cerebrospinal fluid. PMID- 11501818 TI - Two-dimensional mechanical model of the upper end of the humerus. PMID- 11501819 TI - Osteomyelitis of the patella. AB - Osteomyelitis of the patella is an uncommon infection that is primarily a disease of childhood. The diagnosis frequently is delayed because of its rarity and variable presentation, which may be acute with systemic signs or insidious with mild local signs. The diagnosis should be considered in patients with persistent peripatellar pain and swelling, cellulitis, septic prepatellar bursitis, and septic arthritis of the knee that does not respond to standard treatment. Treatment is identical to the treatment of osteomyelitis at other locations. The outcome is favorable in the child regarding range of motion and activity, although there may be persistent changes radiographically. PMID- 11501820 TI - Patellar tumors. AB - Pain in the anterior knee has become synonymous with patella chondromalacia. However, pain also is the first sign of a tumor of the patella. Tumors of the patella are rare, but they often are mistaken for other benign lesions. Benign tumors of the patella are more frequent than malignant tumors (73% versus 27%). The most common benign neoplasms are giant cell tumor and chondroblastoma. Margin of the lesion, cortical involvement, trabecular pattern, and type of the matrix are important features for a radiographic diagnosis. Bone scans, computed tomography scans, and magnetic resonance images are used for staging of the tumor before surgery. Curettage only should be considered for patients with Stage 1 or Stage 2 benign lesions. Patellectomy is indicated for patients with Stage 3 aggressive benign tumors and for patients with Stage IA malignant tumors. A patellectomy with excision of involved soft tissues is used in patients with Stage IB tumors, whereas in patients with Stage IIA tumors, an extensive patellectomy is indicated. In patients with Stage IIB tumors, an extraarticular resection of the knee is preferred when there is a small soft tissue involvement, but amputation is necessary when there is a large soft tissue mass. PMID- 11501821 TI - Surgical repair of cartilage defects of the patella. AB - The structure and biomechanical forces on the patellar joint challenges researchers to define an ideal method for resurfacing the patellar cartilage. The articular surface of the patella presents variability between individuals, and has various minor articulations that bear partial or total compressive, shear, and combined forces during movement. Surgical techniques for the repair of patellar cartilage defects have evolved from cumulative advances in basic science and technology. Such surgeries include the techniques that promote either fibrocartilage formation or hyalinelike cartilage formation. Techniques promoting the formation of fibrocartilage yield short-term solutions because fibrocartilage lacks the durability and the mechanical properties of articular hyaline cartilage. Currently, there is no ideal method for the repair of patellar cartilage defects; all methods are considered experimental. Additional controlled and randomized clinical studies with large series of patients and long-term followup are required. PMID- 11501822 TI - The etiology of chondromalacia patellae. 1961. PMID- 11501823 TI - Alterations in the patella after a high tibial or distal femoral osteotomy. AB - Knee osteotomies realign the knee in an attempt to better distribute forces across the knee. The anatomic and physiologic function of the extensor mechanism, which includes the quadriceps tendon, patella, and patella ligament, may be altered during this procedure. An understanding of these changes is important especially when additional surgery becomes necessary, such as a conversion to a total knee arthroplasty. The current authors discuss patella mechanics and changes in the patella associated with osteotomies about the knee and the influence on normal patella biomechanics. Although patella changes are uncommon after a distal femoral osteotomy, poor total knee arthroplasty outcomes after a high tibial osteotomy attributable to patella alterations exist. Surgical technique during the primary high tibial osteotomy and the conversion to the total knee arthroplasty can reliably improve the final outcome. Rigid internal fixation with early knee mobilization after high tibial osteotomy reduces the incidence of patella baja and improves total knee arthroplasty outcome after a high tibial osteotomy, whereas while patella changes after a distal femoral osteotomy are minimal and largely ignored. PMID- 11501824 TI - Arthroscopic-assisted percutaneous stabilization of patellar fractures. AB - The authors did percutaneous, arthroscopic-assisted osteosynthesis of patellar fractures associated with skin abrasions and lacerations in 11 patients. Skin problems did not delay the operation and the rehabilitation. The average followup was 2.8 years. All fractures healed without complications and good results were obtained in all patients using the criteria of Levack et al. This technique avoids the problems of patellar fractures with skin abrasions and may be adapted to other fractures of the patella. PMID- 11501825 TI - Patella in total knee arthroplasty. AB - The patella is a reliable guide to the success or failure of a total knee replacement. Patients who do not experience peripatellar symptoms or a patellar complication usually have a successful result. Conversely, peripatellar symptoms or complications usually reflect an underlying problem with surgical technique, component designs, or both. Current designs still do not replicate normal kinematics, and current instrumentation and techniques significantly alter the anatomy of the patellofemoral articulation in a substantial percentage of patients. Reproducing extensor mechanism balance and using components that provide adequate congruency and contact area through a physiologic arc of motion should lead to a successful result with minimal patellar symptoms or complications whether or not the patella is resurfaced. Attempting to achieve normal patellofemoral kinematics and minimize patellar complications has led to a better understanding of total knee arthroplasty. PMID- 11501826 TI - Patellectomy: an overview with reconstructive procedures. AB - Patellectomy is a relatively old procedure and its effect on knee function has been a matter of controversy. Although indications for a patellectomy have been narrowed, patellectomy still is indicated as the last choice of treatment in certain situations including comminuted fractures, advanced chondromalacia or osteoarthritis, dislocations, infections, and tumoral conditions. Excision of the patella results in a decrease in the moment arm of the extensor mechanism, alteration of the forces acting on the tibiofemoral joint and instant center of motion, limitation of range of motion, anterior instability, loss of protection of the trochlea from injury, and poor cosmesis. A good technique of patellectomy should overcome all or at least some of these problems. Although comparison of various techniques of patellectomy is impossible because of the lack of prospective randomized trials, reinforcement techniques seem more logical than simple repair procedures. PMID- 11501827 TI - Response of peripheral lymphocytes from patients with ossification of posterior longitudinal ligament. AB - The in vitro response of peripheral blood mononuclear cells or enriched CD4+ T cells from patients with ossification of the posterior longitudinal ligament to anti-CD3 monoclonal antibody has been studied. The response in both was significantly lower in patients with the continuous-type ossification than in patients with the segmental-type ossification and in healthy volunteers, and was inversely correlated with the number of vertebral bodies with ossified ligament. In patients with the segmental-type ossification, the response of peripheral blood mononuclear cells was significantly lower than that in healthy volunteers, but that of the enriched T cells was not. B cell proliferation in response to fixed Staphylococcus aureus cells was significantly lower in patients with the continuous-type ossification than in healthy volunteers but was not correlated with the number of vertebral bodies with ossified ligament. The B cell response in patients with the segmental-type ossification was not lower than that in healthy volunteers. Serum concentrations of transforming growth factor-beta1 and basic fibroblast growth factor also were higher in patients with the continuous type ossification than in patients with the segmental-type ossification and in healthy volunteers. The findings raise the possibility that continuous-type ossification of posterior longitudinal ligament might develop differently from segmental-type ossification. PMID- 11501828 TI - Intracranial subdural hematoma after puncture of spinal meningeal cysts. AB - A patient is reported with an intracranial subdural hematoma after puncture of spinal meningeal cysts. In this case, spinal meningeal cysts were diagnosed by myelography. No intracranial subdural hematoma was detected immediately after myelography. Deterioration in the patient's level of consciousness occurred after puncture of the cysts. The authors speculated that the cerebrospinal fluid pressure dropped rapidly when the spinal meningeal cysts were punctured. This displaced the cerebral bridging veins downward, tearing them and resulting in an intracranial subdural hematoma. PMID- 11501829 TI - Applied biomechanics of the patella. AB - Although numerous prominent orthopaedists of the twentieth century considered the patella to be useless, even detrimental, it now is clear that the patella serves an important biomechanical function. It is a complex lever that magnifies the moment arm of the extensor mechanism. The patellofemoral contact area (the fulcrum of the lever) shifts along a proximodistal axis through the knee's arc of motion. As the knee flexes, the force within the patellar tendon diminishes relative to that of the quadriceps tendon. One's interpretation of patellar tracking is dependent on the choice of coordinates. When assessing tracking by way of anatomic coordinates, patellas are seen to be slightly lateralized at 0 degrees flexion and to follow similar paths down the trochlea. The tracking pattern is the result of an elaborate interplay between the quadriceps muscles, patellofemoral ligaments, the geometry of the trochlea, and the quadriceps angle. The articular cartilage of the patella is the thickest in the human body and does not follow the contour of the subchondral bone. Patellar cartilage is softer and more permeable than that of the trochlea. It is insensate. In size, nature, and number, the facets of the patellar articulation vary from person to person. PMID- 11501830 TI - Nucleus pulposus allograft retards intervertebral disc degeneration. AB - Autogenous implantation of nucleus pulposus or nucleus pulposus cells that were activated by coculture retards intervertebral disc degeneration, but harvesting such grafts causes disc degeneration at the donor site. This study examined whether nucleus pulposus allografts similarly retard disc degeneration and whether such allografting induces immunologic rejection. Japanese White rabbits served as donors and recipients for allografts. Lumbar disc degeneration was induced by aspirating the nucleus pulposus. Two weeks later, intact nucleus pulposus or nucleus pulposus cells were injected and compared with a sham procedure and normal control. The recipients' discs were examined histologically and immunologically at intervals for 16 weeks. Discs receiving an intact nucleus pulposus showed the least degeneration, followed by discs receiving nucleus pulposus cells, both of which were better than no treatment. These findings correlated directly with the intensity of immunochemical staining for Type II collagen. Allogeneic grafts did not induce any appreciable host-versus-graft response. Injection of nucleus pulposus and nucleus pulposus cells retards intervertebral disc degeneration. However, injection of intact nucleus pulposus is more effective than injection of nucleus pulposus cells alone. The intercellular matrix plays an important, but poorly understood, role in preserving intervertebral discs. PMID- 11501831 TI - Radioguided-surgery of early breast lesions. AB - BACKGROUND: Radioguided-surgery has been recently proposed in patients with clinically occult breast lesions. This study aimed to evaluate the feasibility of correctly locating and eradicating, by a single intralesional injection of a radiotracer, any breast lesion and, in the case of malignancy, to perform simultaneous sentinel lymph node (SLN) biopsy procedure. PATIENTS AND METHODS: Sixty-three women with early breast lesions were enrolled: 42 were invasive carcinomas, 16 in situ ductal carcinomas (DCIS) and 5 fibroadenomas. RESULTS: Scintigraphic images clearly identified the lesions in all patients while SLN/s were evident in 88% of them. At surgery all the breast lesions were easily radiolocalized and eradicated with minimum surgical trauma and, for those patients with invasive carcinomas, the SLN technique was performed in 86% of them. No skip metastases were found. CONCLUSION: A single intralesional administration of radiotracer is an easy and reliable procedure to simultaneously locate and remove both the non-palpable breast lesion and the SLN when primary malignancy was intraoperatively confirmed. PMID- 11501832 TI - Serial quantitative PCR analysis of bone marrow samples from breast cancer patients to monitor systemic micrometastases. AB - BACKGROUND: We have previously developed a quantitative calibrated PCR assay to measure cytokeratin 19 (CK19) expression in haematopoietic tissue in order to detect systemic micrometastases. PATIENTS AND METHODS: Serial measurements of CK19 expression in bone marrow of patients with primary breast cancer were performed at operation, at 3 weeks and 6 months postoperatively. RESULTS: Reference values for CK19 expression were established by analysing bone marrow samples from 48 healthy female volunteers or patients without epithelial cancer. Samples from breast cancer patients with CK19 values above the upper reference limit were considered positive. Bone marrow samples taken at operation were positive in 29 out of 141 patients (20.6%) and remained positive in 12, turned negative in 4 and were unavailable in 13 at 6 months postoperatively. CONCLUSION: Serial measurements increase the reliability of detecting micrometastases perioperatively. Further studies are in progress to evaluate the relationship between elevated CK19 values and clinical outcome. PMID- 11501833 TI - Retrospective analysis of administration of a combination of docetaxel and carboplatin for advanced non-small cell lung cancer. AB - Retrospective analysis was performed for 60 patients with advanced non-small-cell lung cancer (NSCLC) who had been treated with combination therapy combining 60 mg/m2 of docetaxel with carboplatin in the range of 200 to 360 mg/m2 (average: 290 mg/m2) every 3 weeks. Considering the patients' performance status, the dose of carboplatin was lowered accordingly, being equivalent to AUC 1.9 to 6.1 (average: 3.26) by the Chatelut formula. The mean treatment cycle was 2.3 (range 1 to 7). Complete response and partial response were observed in 2 and 18 (37.0%) of the 54 evaluable patients, respectively, with a median survival time of 12.8 months and 1-year survival of 56.4%. The calculated AUC of carboplatin was not proportional to the response rate. Moderate myelosuppression was exhibited. The severity of leukopenia increased in relation to the AUC of carboplatin (R2=0.1093), whereas the relation between the platelet count and the AUC of carboplatin was relatively disproportional (R2=0.0553). Although gastrointestinal toxicity was slight, its severity increased dependent on the AUC of carboplatin. No occurrence of neurotoxicity was observed. Treatment with a combination of docetaxel and low-dose carboplatin seemed to be effective and safer in patients with advanced NSCLC. PMID- 11501834 TI - Apoptosis and p53 overexpression in human rectal cancer; relationship with response to hyperthermo-chemo-radiotherapy. AB - Hyperthermo-chemo-radio (HCR) therapy has been found to be effective for rectal cancer. Biomarkers for predicting the effect of HCR therapy are important in determining optimum treatment regimens. Hyperthermo-chemo-radiotherapy (HCR therapy), consisting of hyperthermia at 42 degrees C to 45 degrees C for 40 minutes (twice per week for two weeks), a total of 60 Gy irradiation and administration of 1-hexylcarbamoyl-5-fluorouracil (HCFU) (total 8400 mg), were prescribed pre-operatively for 29 patients with rectal cancer, using tissue specimens collected at pre-treatment biopsy. Apoptosis and overexpression of p53 protein were investigated histopathologically and immunohistochemically. On termination of HCR therapy, all the tumors were surgically resected and effectiveness of the therapy was evaluated histologically. Spontaneous apoptosis was evident in the pre-treatment cancer tissues of 14 patients (48.2%). In this apoptosis-positive group, the positive rate of expression of the p53 protein (21.4%, 3 out of 14) was lower as compared to findings in the apoptosis-negative group (66.7%, 10 out of 15). The response to HCR therapy was better in the apoptosis-positive group than in the apoptosis-negative group. We propose that spontaneous apoptosis is closely related to the function of wild-type p53 protein and is also a predictive biomarker of the effect of HCR therapy for patients with rectal cancer. PMID- 11501835 TI - Mandibular metastatic hepatocellular carcinoma: report of a case involving severe and uncontrollable hemorrhage. AB - Hepatocellular carcinoma (HCC) is a malignant tumor with a marked tendency to spread through the portal system. Metastases from HCC usually involve lungs, surrenal glands, the skeletal and gastroenteric systems, spleen, heart and kidneys. Secondary localizations to the mandible are rare. Generally, bone metastases from HCC appear as osteolytic lesions more likely localized to the ribs, spine, femor, omer, sternum, and then to the mandible. Mandibular metastatic HCC is hemorrhagic in nature because of its hypervascularity. Any diagnostic maneuver that could end in bleeding should be avoided. Non-invasive diagnostic procedures such as computer tomography (CT) scan should be preferred. Among the invasive diagnostic procedures, only fine needle biopsy should be attempted and palliative radiotherapy could be useful for the control of local symptoms. A case report of a hemorrhagic mandibular metastatic HCC that had to be treated surgically, in order to control the severe and profuse bleeding, is presented. PMID- 11501836 TI - Pancreatic acinar cell carcinoma with endocrine differentiation: immunohistochemical and ultrastructural analyses. AB - The majority of pancreatic malignant tumors are adenocarcinomas of the ductal type (ductal cell carcinomas) and combined tumors consisting of different tumor components are very rare. We present here a rare case of acinar cell carcinoma with apparent foci of endocrine differentiation. A 46-year-old man underwent pylorus-preserving pancreatoduodenectomy under the diagnosis of pancreatic tumor. The pancreatic tumor was mainly composed of typical acinar cell carcinoma, but some tumor cells were positive for both acinar and endocrine cell markers such as pancreatic amylase, trypsin, lipase and chromogranin A. At the electron microscopic level, the tumor cells were seen to have numerous electron-dense neuroendocrine, as well as a few zymogen-like, granules. The tumor part positive for both acinar and endocrine cell markers originated from a subclone (dis differentiated tumor cells) of the typical acinar cell carcinoma tissue of the pancreas. PMID- 11501837 TI - Immunohistochemical TRF1 expression in human primary intracranial tumors. AB - BACKGROUND: The telomeric-repeat binding factor (TRF1) participates in a physiological homeostatic mechanism controlling telomere shortening by inhibiting telomerase activity: down-regulation of TRF1 expression results in telomere elongation and may be involved in cell immortalization. PATIENTS AND METHODS: To determine the TRF1 expression by immunohistochemistry (IHC) in human brain tumors, a cohort of 20 consecutive flash-frozen surgical specimens (14 meningiomas and 6 anaplastic astrocytomas (AA)) were collected. RESULTS: Variable levels of TRF1 expression in 12 out of the 14 (87.5%) meningioma samples were observed. By contrast, no expression of TRF1 in tissue samples from AA (p = 0.008) was detected. Positive TRF1 cells were usually more differentiated (less atypical features) and Ki67 negative (inverse statistical association, chi2 = p < 0.001). CONCLUSION: We demonstrated, for the first time, that routine IHC techniques are capable of identifying TRF1 expression in intracranial tumors, which is heterogeneously expressed in meningiomas, but absent in AA. Although these preliminary observations need confirmation from larger studies, the TRF1 status in intracranial tumors might become of prognostic value. PMID- 11501838 TI - Cyclooxygenase-2, P-glycoprotein-170 and drug resistance; is chemoprevention against multidrug resistance possible? AB - BACKGROUND: It is generally accepted that P-glycoprotein 170 (MDR1/Pgp170) expression in breast tumors results in poor response to chemotherapy due to its ability to export chemotherapeutic agents. Studies indicate that the use of non steroidal anti-inflammatory drugs (NSAIDs) may enhance the anti-tumor activity of cancer chemotherapeutic agents and reduce the risk of many cancers. The best known function of NSAIDs is to block the enzyme cyclooxygenase (Cox), the rate limiting enzyme in the conversion of arachidonic acid to prostaglandins. In this study we investigated whether expression of the inducible isoform of Cox (Cox-2) is linked with the multidrug resistance phenotype in breast cancer. METHODS: Expression of Cox-2 and MDR1/Pgp170 was investigated in tumor specimens along with normal epithelium in breast cancer patients using immunohistochemisrty. Expression of Cox-2, MDR1/Pgp170, Protein Kinase C (PKC), and Activator Protein 1 (AP1) were investigated in a series of increasingly resistant human MCF-7 breast cancer cells compared to wild type using immunohistochemistry, Western blots, Northern blots, RT-PCR, and Southern blots. RESULTS: Immunohistochemical analyses of human breast tumor specimens revealed a strong correlation between expression of Cox-2 and MDR1/Pgp170. In drug resistant cell lines that over-express MDR1/Pgp170 there was also significant up-regulation of Cox-2 expression. In addition, PKC and AP1 subunits c-Jun and c-Fos were also upregulated. We hypothesized that increased prostaglandin production by Cox-2 induces PKC and the expression of transcriptional factor c-Jun, which in turn, induces the expression of MDR1/Pgp170. CONCLUSION: We propose that pretreatment with selective Cox-2 inhibitors may be useful in the prevention of multidrug resistance in response to cancer chemotherapy and should be further evaluated. PMID- 11501839 TI - p53, bcl-2 and thymidine phosphorylase as predictive markers of chemotherapy in patients with advanced and recurrent gastric cancer. AB - PURPOSE: To study the relationship between expression of p53, bcl-2, thymidine phosphorylase and Ki-67 and the response to chemotherapy and survival in patients with recurrent and advanced gastric cancer. MATERIALS AND METHODS: Protein expression was assessed immunohistochemically in 28 patients treated with 5 fluorouracil, pirarubicin and cisplatin (FAP). RESULTS: The response rate in patients positive for p53 expression was 23% compared with 47% of p53-negative patients. The response rate was also reduced from 44% in patients negative for bcl-2 protein expression to 25% in bcl-2 positive patients. Thymidine phosphorylase (dThdPase) expression was observed in 20 patients (71%), 50% of whom responded to chemotherapy, while patients negative for dThdPase expression did not respond to chemotherapy. The correlation between response rate and dThdPase-positivity was statistically significant (p < 0.05). However, with regard to patient survival, p53- and bcl-2-negative patients showed significantly better survival than patients positive for p53 and/or bcl-2 (p = 0.036). CONCLUSION: While dThdPase expression may be a useful predictor of response to chemotherapies that include 5-FU, p53 and bcl-2 expression may predict the outcome of patients with recurrent and advanced gastric cancer following chemotherapy. PMID- 11501840 TI - Human herpesviruses HHV-4 (EBV) and HHV-6 in Hodgkin's and Kikuchi's diseases and their relation to proliferation and apoptosis. AB - BACKGROUND: Human herpesviruses types 4 and 6 (EBV, HHV-6) are frequently found in Hodgkin 's disease (HD) and--to a certain extent--in Kikuchi-Fujimoto's disease (KFD). Both viruses are apparently related to proliferative and/or apoptotic processes as represented by HD or KFD respectively. OBJECTIVE: To correlate frequency and location of antigen- and DNA expression of both viruses in HD and KFD tissue sections in relation to markers for cell proliferation and apoptosis. STUDY DESIGN: Archival lymph node biopsies from 103 patients with HD and 14 KFD patients were investigated immunohistologically for viral antigen expression (EBV LMP- 1: HHV-6 pl 10/60), Ki67/PCNR, marker for proliferation (MIB1)/p53 and WAF1 for apoptosis. Viral DNA was shown by in situ hybridization. Apoptosis was determined by ISEL and TUNEL techniques. RESULTS: HD is frequently infected by both EBV and HHV-6 while KFD tends to be infected only by HHV-6. EBV in HD is present in HD cells and in Reed-Sternberg cells (HD/RS cells), HHV-6 preferentially in lymphocytes and in histiocytes in both HD and in KFD. Proliferation marker Ki67 is found in lymphocytes and histiocytes of both diseases and in HD and RS cells in HD. Apoptosis is demonstrated in lymphocytes and histiocytes preferentially in KFD and to a lesser extent also in HD. CONCLUSION: Although EBVand HHV-6 may not be openly oncogenic in HD and KFD, they may well influence the course of the disease. Dual infection in HD appears to support proliferative processes, i.e. a predominance of EBV effects. Single infection with HHV-6 in KFD instead appears to favor an apoptotic course. These effects are--according to the literature--possibly cytokine-mediated. PMID- 11501841 TI - Tie-1 tyrosine kinase is an independent prognostic indicator for invasive breast cancer. AB - Receptor tyrosine kinases are known to be involved in the growth, progression and metastasis of solid tumors. We investigated the relationship between tie-1 expression and progression of invasive ductal breast carcinoma with immunohistochemical analysis. Tie-1 protein was detected in the microvessel endothelial cells and cytoplasm of tumor cells. The tumor size and stage were significantly associated with the expression of tie-1, which portends a worse 5 year disease-free status (39.3% v 59.2%, p = 0.07) and overall survival rate (67.3% v 93%, p = 0.02) than those without tie-1 expression. Multivariate analysis demonstrated that larger tumor size, presence of lymph node metastasis and tie-1 expression were independent prognostic parameters, both in 5-year disease-free survival and overall survival. Patients with lymph node metastases and tie-1 expression had the worst 5-year disease-free survival (0%) and overall survival (42.4%) compared to those without tie-1 expression (50.2%, 85%). In lymph node negative patients, those without tie-1 expression had better 5-year disease-free survival and overall survival (72.9%, 100%) compared to those with tie-1 expression (65.5%, 87.7%). We conclude that tie-1 expression is an independent prognostic factor for invasive ductal breast carcinoma, adversely affecting survival of breast cancer patients with positive nodes to a significant extent. PMID- 11501842 TI - Human alpha-and beta-defensin immunoreactivity in oral mucoepidermoid carcinomas. AB - The purpose of this study was to demonstrate the immunohistochemical localization and distribution of human alpha- and beta-defensins, peptides with antimicrobial activity, in oral mucoepidermoid carcinoma tissue. Tissue samples were embedded in paraffin and alpha- and beta-defensins were immunostained by the streptavidin biotin coupled peroxidase method. Cancer cells that constituted the ducts, as well as neutrophils, were positively immunostained with the anti-alpha-defensin antibody (HNPs). On the other hand, epidermoid cells and intermediate cells were intensely stained with the anti-beta-defensin-2 (HBD-2) antibody. Mucous secreting cells were clearly not immunostained with the anti-HBD-2 antibody. The epithelial hyperplasia region adjacent to the tumor tissues was also positively immunostained with the anti-HBD-2 antibody. PMID- 11501843 TI - Diagnostic application of CD44 variant expression in pancreatic juice for detection of pancreatic neoplasm. AB - Recently, increased and disorganized expression of CD44 variant exons (CD44v) has been demonstrated in several types of human malignancy. We tried to investigate CD44v expression in pancreatic juice from patients who underwent endoscopic retrograde pancreatography. We analyzed 24 patients with pancreatic neoplasms diagnosed histologically (adenocarcinoma, 17; adenoma, 7) and 15 patients with non-neoplastic lesions. The expression of CD44v mRNA in pancreatic juice was detected by using the reverse-transcription polymerase chain reaction technique followed by Southern hybridization with exon-specific probes. Of 17 patients with adenocarcinoma, 14 (82%) showed expression of CD44v6 mRNA and 11 (65%) showed expression of CD44v2 mRNA. Of 7 patients with adenoma, 6 (86%) were positive CD44v6 mRNA expression and 2 (29%) for CD44v2 mRNA expression; while, out of 15 patients with non-neoplastic lesion, 5 (33%) showed positive findings for CD44V6 mRNA and 3 (20%) for CD44v2 mRNA. Comparing of diagnostic accuracy among CD44v6, CD44v2 and cytological examination, the sensitivities for adenocarcinoma were 82%, 65% and 41% respectively. However, the specificity was lower in CD44v6 (50%), CD44v2 (77%) than in cytology (100%), because CD44v was positive in adenoma cases and normal cases. A combination of RT-PCR analysis for the expression of CD44v with cytological examination in the pancreatic juice may increase the accuracy of diagnosis for pancreatic cancer. PMID- 11501844 TI - Prognostic value of beta1-integrin (=CD29) in serous adenocarcinomas of the ovary. AB - BACKGROUND: Integrins are heterodimeric transmembranous proteins, comprised of two transmembrane subunits, called alpha and beta. They bind to various structures of the extra cellular matrix (ECM) and are responsible for cell-cell interactions. The aim of the current study was to evaluate the prognostic value of beta1-integrin in patients suffering from malignant serous surface epithelial stromal tumors of the ovary. MATERIALS AND METHODS: With immunohistochemical methods we investigated 76 formalin-fixed, paraffin-embedded tissue samples. FIGO stages I, II, III and IV were present in 18, 10, 41 and 7 cases, respectively. RESULTS: 12 sections (15.8%) stained positive for beta1-integrin (=CD29), 64 sections (84.2%) showed no expression of beta1-integrin. The product limit method by Kaplan and Meier showed no statistical significance of beta1-integrin expression on overall survival, (p = 0.1005, log-rank test). Also the univariate Cox regression analysis showed no statistical significance of beta1-integrin expression on overall survival (p=0.1066), whereas in the multivariate analysis, adjusted for grading, FIGO stage and residual tumor, the expression of beta1 integrin turned out to be significantly correlated with reduced overall survival (p=0.0208). CONCLUSION: Our results using multivariate Cox regression analysis indicated a shorter overall survival for patients with positive staining for beta1-integrin in malignant serous surface epithelial-stromal tumors of the ovary. This pilot study warrants further investigation of the role of beta1 integrin in ovarian cancer. PMID- 11501845 TI - Expression of mucin-associated carbohydrate core antigens in esophageal squamous cell carcinomas. AB - In contrast to gastrointestinal cancer, where a correlation between the expression of different mucin-associated core antigens with clinico-pathological parameters and survival probability, has been established, little is known about their importance in esophageal cancer. Therefore, we characterized esophageal squamous cell carcinomas from 84 patients immunohistochemically by applying monoclonal antibodies (mabs) directed against the Thomsen-Friedenreich (TF) antigen MUC1-bound TF antigen and sialyl-Tn. TF was observed in about 40% of the cases and MUC1-TF epitope in about 75%. Sialyl-Tn was detectable in about half of the carcinomas under study. None of these mabs showed any correlation between binding pattern and clinico-pathological variables, such as TNM stage, lymph node metastasis or grading. However, a strong expression of MUC1-TF epitope as well as sialyl-Tn antigen predicted a poor survival probability. In conclusion, it is suggested that mucin-associated carbohydrate core antigens are involved in the biology and clinical course of esophageal squamous carcinomas. PMID- 11501846 TI - Hepatocyte growth factor production by peripheral blood mononuclear cells of recurrent cancer patients. AB - The elevation of circulating hepatocyte growth factor (HGF) concentrations is associated with tumor progression and prognosis in cancer patients. We aimed to clarify whether peripheral blood mononuclear cells (PBMC) of cancer patients can produce circulating HGF. In sera and PBMC supematants of 39 cancer patients and 55 control subjects, we measured HGF concentrations by enzyme-linked immunosorbent assay and analyzed HGF expression in PBMC by reverse transcriptase polymerase chain reaction and Western blotting. Compared with primary cancer patients and control subjects, recurrent cancer patients showed higher serum HGF concentrations (p < 0.05, p < 0.01 respectively), higher HGF concentrations in PBMC supernatants (p < 0.01 for both) and more frequent HGF mRNA expression in PBMC. Additionally, HGF mRNA was induced in PBMC of a control subject cultured with the sera of recurrent cancer patients. These results indicate that PBMC of recurrent cancer patients can produce HGF and that their sera may contain substances that induce HGF mRNA expression. PMID- 11501847 TI - Sestamibi scintimammography in pT1 breast cancer: alternative or complementary to X-ray mammography? AB - The aim of this study was to compare the results obtained using X-ray mammography (MG) and 99mTc-sestamibi scintimammography (SSM) in patients with breast lesions < or = 2 cm. One hundred and thirty-four women (median age 52 years, range 32 78), who had already been selected for open breast biopsy, underwent both MG and SSM prior to surgery. Final pathology showed 27 (20.1%) benign breast lesions and 107 (79.9%) breast cancers (pT1a=5 [4.7%], pT1b=50 [46.7%], pT1c=52 [48.6%]). The sensitivity, positive predictive value, negative predictive value and accuracy were 81.3%, 97.6%, 55.6% and 83.6% for SSM and 83.2%, 89.9%, 48.6% and 79.1% for MG, respectively. The results were similar (p=NS), but patients with breast cancer (BC) undetected by MG were significantly (p<0.05) younger than those in which the tumor was correctly diagnosed, while the age did not affect SSM sensitivity. SSM was more (p<0.05) specific than MG in BC detection both in the overall group of patients (92.6% vs 63.0%) and in those with < or = 10 mm breast lesions (94.7% vs 63.2%). In conclusion, in patients with suspected BC sized < or = 2 cm, SSM may help in surgical planning because of its high specificity, and should be considered complementary to MG, especially in younger women. PMID- 11501848 TI - HER-2/neu overexpression in breast cancer: an immunohistochemical study including correlations with clinicopathologic parameters, p53 oncoprotein and cathepsin-D. AB - PURPOSE: To evaluate the relationship between HER-2/neu overexpression and standard as well as investigational prognostic factors in Greek females with invasive breast carcinoma. MATERIALS AND METHODS: Paraffin-embedded tumor sections from 128 consecutive primary breast cancer patients were screened for HER-2/neu oncoprotein (p185) overexpression by immunohistochemistry using the polyclonal antibody A 0485. The relationship between HER-2/neu staining and other established markers of prognosis were examined using both a univariate and a multivariate analysis. RESULTS: HER-2/neu overexpression was detected in 46.1% of tumors. No statistical correlation was found between HER-2/neu and age, tumor diameter, histologic type, nodal status, tumor grade, stage, estrogen and progesterone receptor status or cathepsin-D. A significant correlation was noted between HER-2/neu and p53 overexpression in the total sample (p=0.014) as well as in node-positive patients (p=0.026). In those groups, HER-2/neu and p53 co expression was found in 19.5% and 21.6% of cases, respectively. In node-negative patients, HER-2/neu overexpression did not correlate with any other marker. CONCLUSION: HER-2/neu overexpression seemed to correlate only with p53 oncoprotein. PMID- 11501849 TI - Comparison of technetium-99m methoxyisobutylisonitrile single photon emission computed tomography and computed tomography to detect recurrent or residual nasopharyngeal carcinomas after radiotherapy. AB - The diagnostic accuracy of technetium-99m methoxyisobutylisonitrile (Tc-MIBI) single photon emission computed tomography (SPECT) of head and neck to differentiate between recurrent or residual nasopharyngeal carcinomas (NPC) and benign lesions after radiotherapy was evaluated, and compared with computed tomography (CT). Thirty-six NPC patients 4 months after radiotherapy underwent Tc MIBI SPECT and CT of head and neck, as well as histopathological examination of nasopharyngeal biopsies. Based on the biopsy results, the sensitivity, specificity, and accuracy of CT were 73%, 88%, and 83%, respectively. The sensitivity, specificity, and accuracy of Tc-MIBI SPECT were 64%, 96%, and 86%, respectively. The sensitivity, specificity, and accuracy of combined Tc-MIBI SPECT and CT were 100%, 88%, and 92%, respectively. Tc-MIBI SPECT had a better specificity and a lower sensitivity to differentiate benign lesions from recurrent/residual NPC when compared with CT The combined use of CT and Tc-MIBI SPECT significantly increased accuracy compared with the single use of either Tc MIBI SPECT or CT to differentiate benign lesions recurrent/residual NPC. PMID- 11501850 TI - Radiation therapy for oncological emergencies. AB - AIMS AND BACKGROUND: Radiation therapy holds a fundamental role in oncological emergencies such as superior vena cava syndrome, spinal cord compression and endocranial hypertension. The purpose of our study was, by comparing schedules of treatment, to confirm the efficacy of hypofractionated radiation therapy. METHODS: From January 1994 to December 1998, 43 patients with superior vena cava syndrome, 37 patients with metastatic spinal cord compression and 108 patients with endocranial hypertension secondary to metastasis were treated at our institution. In the group of patients with superior vena cava syndrome, radiotherapy schedules were: 4 Gy x 5 to a total dose of 20 Gy (23 patients) and 3 Gy x 10 to a total dose of 30 Gy (20 patients). In the group of patients with spinal cord compression, radiation schedules were: 3 Gy x 10 to a total dose of 30 Gy (15 patients); 4 Gy x 5 to a total dose of 20 Gy (12 patients); a single fraction of 8 Gy in 10 cases, repeated after 1 week in 7 responder cases to a total dose of 16 Gy. 5 out of 37 patients were underwent to laminectomy plus stabilization of the spine and post-operative radiotherapy. In the group of patients with endocranial hypertension, radiotherapy schedules were: 6 Gy x 2 to a total dose of 12 Gy (53 patients), repeated after 4 weeks in 34 responder patients and 3 Gy x 10 to a total dose of 30 Gy (55 patients). RESULTS: The patients with superior vena cava syndrome, revaluated after 4 weeks at the end of treatment, obtained a partial remission of symptomatology in 73.9% with 20 Gy and in 75% with 30 Gy. The patients with spinal cord compression obtained symptomatic relief in 73.3% with 30 Gy, in 66.6% with 20 Gy and in 70% of cases treated with 8 Gy. The patients with endocranial hypertension obtained symptomatic relief in 64.1% with 12 Gy and in 63.3% with 30 Gy. CONCLUSION: Histology, pretreatment and performance status were important prognostic factors for the response to therapy. Our results demonstrated no significant difference among different schedules of radiotherapy and confirmed the importance of radiotherapy for oncological emergencies: it improves the quality of life and, in responding patients, is associated with a longer survival time. PMID- 11501851 TI - From codeine to transdermal fentanyl for cancer pain control: a safety and efficacy clinical trial. AB - BACKGROUND: Fentanyl is a synthetic opioid, suitable for transdermal delivery, offering an interesting solution as a step 3 opioid in cancer pain treatment. The purpose of the study was to carefully investigate: 1) the feasibility of the direct conversion from codeine to TTS fentanyl, in patients already receiving codeine and requiring strong opioids for their analgesia; 2) the safety of 25 microg/hour incremental steps and at shorter than 72-hour intervals, if clinically required. PATIENTS AND METHODS: 130 patients were judged eligible for the study. All the patients were receiving 280-360 mg or more of codeine and required strong opioid for their analgesia. The study lasted 56 days. The initial dose was 25 microg/hour. TTS fentanyl for all patients. Data assessments were made on baseline, day 1, day 2, day 3, in the hospital and thereafter on days 7, 14, 21, 28, 42 and 56. After the patch application, all the patients were given an immediate release oral morphine (5 mg) every 4-6 hours for the first 12 hours and then if needed only as rescue doses. The patients remained in the hospital for the first three days of the study where follow-up (pain score, satisfaction, side effects etc.). was recorded by the palliative care team and by daily cards. RESULTS: The itnitial dose of fentanyl was 25 microg/hour while the mean dose on day 3 was 45.9 microg/hour. All the patients required upward titration of the study medication during follow-up visits. On day 56 the mean dose of fentanyl was 87.4 microg/hour. Mean pain intensity decreased from an initial 5.96 on the baseline to 0.83 on day 3. Karnofsky scale measurements between treatment phases revealed non-significant changes. The rate of overall satisfaction was quite high. Nine patients discontinued the study due to inadequate pain relief or side effects between day 7 and day 28, while five patients died between day 28 and day 56. Constipation, nausea and vomiting were the most common side effects. Skin reaction was relatively mild and acceptable during the study. CONCLUSION: Under controlled conditions, TTS fentanyl seems to be feasible for direct conversion from mild to strong opioids and additionally, 25 microg/hour incremental steps day by day can be made by palliative care specialists, if clinically required for cancer pain management. PMID- 11501852 TI - Postoperative radiotherapy after prostatectomy can be associated with severe side effects. AB - This retrospective study was initiated to evaluate the efficacy and side effects of post-prostatectomy external beam radiation therapy (XRT) with a short time interval between surgery and irradiation in patients with prostate adenocarcinoma. Sixteen patients were investigated. The overall results in this study were 3 deaths due to recurring disease and two relapses after an average follow-up of 60 months. Severe side effects were observed. Two patients required surgical intervention due to severe post-radiotherapy side effects. The reason for this could be the high dose delivered to peripheral organs and/or a too short time interval between surgery and postoperative XRT. The results of this study confirmed that postoperative XRT can improve local control frequency in prostate carcinomas. It is recommended that the time interval between surgery and postoperative radiotherapy should to be 3-6 month. PMID- 11501853 TI - Genetic polymorphisms of CYP1A1 and GSTM1 and lung cancer risk. AB - Susceptibility to lung cancer may, in part, be determined by interindividual differences in the cytochrome P450-catalysed bioactivation and the glutathione S transferase-catalysed detoxification of procarcinogens. Therefore a lung cancer case-control study was set up to investigate the association of three polymorphisms of the CYP1A1 gene (CYP1A1*2A, CYP1A1*2B, CYP1A1*4) and GSTM1*0 genotype with lung cancer risk in Austrian Caucasians. Genomic DNA was isolated from the peripheral blood lymphocytes of 134 male lung cancer patients and 134 age-matched controls with nonmalignant conditions and PCR-based analyses were performed. There was no significant difference in risk between cases and controls, either for the CYP1A1*2A (OR=1.09, 95%CI=0.46-2.58), CYP1A1*2B (OR=1.09, 95%CL=0.46-2.58) or for the CYP1A1*4 polymorphism (OR=0.49, 95%CL=0.20 1.16). The prevalence of the GSTM1*0 genotype in the lung cancer group (47.8%) was comparable to that found in the control group (49.3%) and also had no effect on lung cancer risk (OR=0.94, 95%CL=0.54-1.57). Further, in a subgroup of male ever-smokers (n=126), no significant influence on the relative risk was found for these polymorphisms. Our results suggest that these investigated polymorphisms can not be considered as genetic susceptibility markers for lung cancer within the Austrian Caucasian population. PMID- 11501854 TI - Melphalan monitoring during hyperthermic perfusion of isolated limb for melanoma: pharmacokinetic study and 99mTc-albumin microcolloid technique. AB - BACKGROUND: The kinetics of melphalan leakage from extracorporeal fluid to the peripheral blood was studied in ten patients undergoing hyperthermic isolation perfusion of the lower limbs as an adjuvant treatment in high-risk melanoma. MATERIALS AND METHODS: Systemic leakage was monitored by a new technique using 99mTc-albumin microcolloid. Serial samples were drawn from a peripheral vein and from the perfusion circuit during surgical treatment and analysed by HPLC. RESULTS: The leakage measured with 99mTc-albumin microcolloid ranged from 1.5 to 18%/h (mean 8%/h). The average concentrations in the perfusate were 200-300-fold those found in the systemic circulation. A good correlation (R=0.945) was obtained between systemic AUC (0 to 1 hour) and leakage measured through the 99mTc procedure. Negligible toxicity was found and the survival rate yielded 92% of objective response. CONCLUSION: By studying the pharmacokinetic data of melphalan in the circuit and in the systemic circulation, we were able to validate the 99mTc procedure used during clinical perfusion. Moreover, considering the efficiency of the system as well as the minimum toxicity and the high survival rate, a reduction of perfusion time may be considered. PMID- 11501855 TI - Nourishing the brain: supporting dietetic research opens doors for the entire profession. PMID- 11501856 TI - Zoo nutrition: a walk on the wild side. PMID- 11501857 TI - Folic acid fortification: informed mothers, healthy babies. PMID- 11501858 TI - Optimizing nutritional health for children through school-based initatives. PMID- 11501859 TI - Breastfeeding and adolescent mothers. PMID- 11501860 TI - The Internet Healthcare Coalition: eHealth Ethics Initiative. PMID- 11501861 TI - Past, present, and future of the Food Guide Pyramid. PMID- 11501862 TI - Development and validation of a screening tool to identify eating disorders in female athletes. AB - OBJECTIVE: To develop and validate a screening tool to identify eating pathology in women athletes. DESIGN: Three cross-sectional subject groups were established: college women athletes diagnosed with eating disorders; college women athletes without eating pathology; college women who did not participate in athletics but were diagnosed with an eating disorder. The Female Athlete Screening Tool (FAST), and 3 valid psychometric measures were administered to subjects in all groups. Internal reliability, discriminant and concurrent validity were established. SUBJECTS/SETTINGS: Women college students between the ages of 18 and 23 years (n=41) from the University of Connecticut and St Joseph College were recruited. The athletes were screened for eating disorders by a sports medicine team. STATISTICAL ANALYSIS: Cronbach's alpha, one-way analysis of variance, and correlation analyses. RESULTS: Reliability analysis indicated a high internal consistency of the FAST (Cronbach's alpha = 0.87). Athletes with eating disorders scored significantly higher on the FAST as compared with athletes without eating pathology and nonathletes with eating disorders (P<.001), which demonstrated discriminant validity. Correlation analyses showed that the FAST was strongly correlated to the Eating Disorder Examination-Questionnaire (0.60, P<.05) and Eating Disorder Inventory (0.89, P<.001). APPLICATIONS/CONCLUSIONS: Early detection of eating disorders can help prevent the onset or severity of a clinical eating disorder. By using the screening tool that has been validated for use in women athletes, dietetic professionals can quickly identify those athletes who need assistance with their aberrant eating habits. PMID- 11501863 TI - Higher carbohydrate intake is associated with decreased incidence of newborn macrosomia in women with gestational diabetes. AB - OBJECTIVE: To study the influence of energy and macronutrient intake on infant birthweight in women with gestational diabetes mellitus undergoing intensive management. DESIGN: This prospective study evaluated the impact of intensive management of gestational diabetes on maternal and fetal morbidity, and addressed the relationship between food intake and infant birthweight. SETTING: Fifteen maternity hospitals in northern France. SUBJECTS: Ninety-nine women with gestational diabetes or gestational mild hyperglycemia diagnosed between 24 and 34 weeks of gestation were surveyed. After 1 was excluded because of a premature birth and 18 were excluded as underreporters, 80 women were included in the final analysis. Diet intake was assessed by a dietary history at the first interview, and by two 3-day diet records at the 3rd and 7th week after diagnosis. RESULTS: In a forward-stepwise regression analysis (controlling for maternal age; smoking; parity; prepregnancy BMI; pregnancy weight gain; gestational duration; infant sex; fasting and 2-hour postprandial serum glucose; insulin therapy; and energy, fat, protein and carbohydrate intake during treatment) infant birthweight was positively associated with gestational duration (beta = +0.34, P<.002), and negatively with smoking (beta = -0.27, P<.02) and carbohydrate intake (beta = 0.24, P<.03). There were no large-for-gestational-age infants among women whose carbohydrate intake exceeded 210 g/day. CONCLUSION: For women with gestational diabetes undergoing intensive management, higher carbohydrate intake is associated with decreased incidence of macrosomia. APPLICATION: These findings suggest that nutrition counseling in gestational diabetes must be directed to maintain a sufficient carbohydrate intake (at least 250 g per day), which implies a low-fat diet to limit energy intake. A careful distribution of carbohydrate throughout the day and the use of low-glycemic index foods may help limit postprandial hyperglycemia. PMID- 11501864 TI - Nutritional assessment of pregnant women enrolled in the Special Supplemental Program for Women, Infants, and Children (WIC). AB - OBJECTIVE: To evaluate nutrient intake from dietary sources for 95 pregnant women enrolled in the Special Supplemental Program for Women, Infants, and Children (WIC). SUBJECTS/SETTING: Women were recruited from Minneapolis and St Paul area WIC clinics between January and June 1999. Based on estimates from the WIC clinics, the study was described to 159 (63%) of the 251 potentially eligible women. Of these 159 women, 107 (67%) completed the in-person interview. Ninety five (89%) were included in the nutrition analysis. METHODS: Each woman completed a 1-hour in-person interview that included a questionnaire, anthropometric measurements, and a venous blood sample. A shortened Block 98 food frequency questionnaire was used to assess dietary intake. Serum ferritin was measured for 86 women. Means, medians, and standard deviations of dietary intake were explored as well as body mass index distributions. Additionally, the percentage of women consuming less than two thirds of the recommended dietary allowance (RDA) for certain nutrients was calculated. RESULTS: Overall, the women reported consuming only 85% of the RDA for energy. The average percentage of energy from fat was higher than recommended (37% vs 30%). The most notable nutrient shortfall was iron; 90% of the women reported consuming less than 2/3 of the RDA. Additionally, serum ferritin analysis classified 22% of the women with iron-deficiency anemia (<12 mg/L). CONCLUSIONS: Interventions should be designed to decrease fat consumption and increase iron intake among pregnant WIC participants to meet dietary recommendations. PMID- 11501865 TI - Point-of-purchase messages framed in terms of cost, convenience, taste, and energy improve healthful snack selection in a college foodservice setting. AB - OBJECTIVE: To examine the effects of a point-of-purchase (POP) intervention emphasizing various properties of healthful food items on college students' snack purchases. DESIGN: In Study 1, vegetable baskets (containing cut pieces of vegetables), fruit baskets (containing cut pieces of fruit), pretzels, and yogurt were promoted in separate POP interventions. Food sales were monitored over 2 week baseline, 4-week intervention, and 2-week follow-up periods. In Study 2, yogurt was promoted across a 2-week baseline, 12-week intervention, and 2-week follow-up periods and an intercept survey was conducted. SUBJECTS/SETTING: Approximately 2,280 university students were potentially exposed to the intervention, and 72 students responded to the intercept survey. INTERVENTION: POP messages were placed on an 11 x 17-in poster located at the cafeteria entrance, and two 4 x 2.5-in signs placed next to the targeted food item. Messages emphasized the Budget-friendly, Energizing, Sensory/taste, Time efficient/convenient (BEST) stimulus properties of food. MAIN OUTCOME MEASURES: Daily sales of the targeted food items. STATISTICAL ANALYSES PERFORMED: Analyses of variance with Tukey post hoc tests were used to compare food sales during the baseline, intervention, and follow-up periods. RESULTS: In Study 1, yogurt and pretzel sales increased during the intervention and post-intervention periods (P<.05). Interventions had no effect on fruit basket and vegetable basket sales (P>.05), but whole fruit sales increased during the fruit basket intervention and follow-up (P<.05). In Study 2, yogurt sales were significantly greater during the intervention and follow-up periods than at baseline (P<.01). APPLICATIONS/CONCLUSIONS: Using the BEST properties in POP interventions may be beneficial in promoting the consumption of healthful foods among university students, particularly when the targeted foods are priced comparably to less healthful foods. PMID- 11501866 TI - College and university students present a challenge for nutritionists. PMID- 11501867 TI - Development of a medical nutrition therapy protocol for female collegiate athletes. AB - Nutrition intervention can improve athletic performance and reduce the risk of nutrition related problems in women athletes. The current healthcare environment demands that dietitians document the outcomes of the medical nutrition therapy (MNT) they provide. This requires the development and validation of MNT protocols so that outcomes can be documented and compared in similar populations across multiple settings. The purpose of this project was to develop a sports nutrition management MNT protocol for collegiate women athletes. A registered dietitian currently working with collegiate women athletes collaborated with four dietitians from the community to develop an MNT protocol. Further testing and validation using this MNT protocol will help dietitians document the outcomes of their interventions in this population. PMID- 11501868 TI - Score points by ducumenting treatment interventions. PMID- 11501869 TI - Post-diagnosis family adaptation influences glycemic control in women with type 2 diabetes mellitus. AB - Family adaptation to type 2 diabetes management after nutrition education was studied as part of a 2.5-year ethnography. Data analysis included descriptive data (observations, transcribed audio recordings, written field notes) and quantitative data (self-monitored morning fasting blood glucose, food records) from the families of 5 New York suburban women. The following inferences emerged from data analysis. Among the women with improved glycemic control, the home and family routines had changed. Diabetes management was observed as certain tasks: food tasks, patient care tasks, management tasks, and medical advice tasks. The social context of the women's daily lives influenced their glycemic control; specifically, family members other than the women with diabetes took on new "hospital roles and routines." Nutrition education should be directed toward assessing existing family social support and helping women organize and supplement this support to ensure the successful completion of the tasks of diabetes self-management. PMID- 11501870 TI - Children's patterns of macronutrient intake and associations with restaurant and home eating. PMID- 11501871 TI - Development of a food and activity record and a portion-size model booklet for use by 6- to 17-year olds: a review of focus-group testing. PMID- 11501872 TI - Fundamental dietary patterns and their correlates among US whites. PMID- 11501873 TI - Wet-nesting of foodservice dishware: investigation and analysis of potential bacterial contamination. PMID- 11501874 TI - Strategies for improving follow-up client appointment-keeping compliance. PMID- 11501875 TI - The new credential: advanced diabetes management. AB - A new multidisciplinary credential, the advanced diabetes manager, has been developed by the American Association of Diabetes Educators (AADE) and the American Nurses Credentialing Center (ANCC), in collaboration with the American Dietetic Association, the American Diabetes Association, and the American Pharmaceutical Association (APhA). The credential, designated as BC-ADM, is conferred upon successful completion of an exam to registered dietitians, registered nurses and registered pharmacists who hold advanced degrees and meet experience requirements. The exam is based on the scope and standards of practice approved by the governing agencies of the sponsoring organizations and defines advanced diabetes management knowledge and skills associated with improved outcomes. Information related to the test content outline, application process, suggested exam preparation, test sites, etc., is available from www.aadenet.org or www.nursingworld.org/ancc. PMID- 11501876 TI - Standards of professional practices for dietetics professionals in management and foodservice settings. PMID- 11501877 TI - Herpetic gingivostomatitis in otherwise healthy adolescents and young adults. AB - The purpose of this study was to review and analyze the records of herpes simplex infections from a specialist Oral Medicine clinic in Iceland, to investigate the clinical impression that the age of patients experiencing initial infection with this virus was higher than expected and that the character of the clinical picture of the disease had changed. Records of patients with herpes infections attending the Oral Medicine clinic covering a 3-year period were examined and the clinical and virological data analyzed. Diagnosis was based on clinical appearance, history, and viral identification with culture or detection of viral DNA by means of the polymerase chain reaction. Records of 60 patients (34 female) were included in the study (mean age, 23.1 years; range, 2 68 years). No patients were known or suspected to be positive for human immunodeficiency virus, none was known to be immunocompromised, and 38 patients (mean age, 16.6 years; 21 female) were diagnosed as having primary herpetic gingivostomatitis. Eighteen patients (mean age, 36.2 years; 11 female) had lesions of recurrent herpes simplex infection present on the oral mucosa. Primary infection with herpes simplex virus was more common in young adults than had been expected. Recurrent infections appeared on the oral mucosal even in otherwise healthy patients, and the clinical course of these infections in this age group sometimes followed a more severe course than that seen in young children. PMID- 11501878 TI - Dental caries and dental health behavior of patients with primary Sjogren syndrome. AB - This comprehensive study was carried out to describe and analyze the oral health status, perceived oral health problems, patients' costs, and oral health behavior in a group of patients with primary Sjogren syndrome (PSS). In particular, the objective of this report was to assess whether Sjogren syndrome patients had more dental caries experience than a control group. Data were collected by means of interviews and clinical oral examinations. The study comprised 53 patients with PSS and a control group of 53 persons matched by age. Among the younger patients the number of decayed, missing, or filled teeth (DMFT) was 22.3, compared with 18.8 among controls (P< 0.05). In parallel, the DMFT in the old-age PSS patients was 26.2, against a DMFT of 22.1 for controls (P< 0.001). On average, the young patients had seven teeth missing, whereas two missing teeth were found among controls (P < 0.01). PSS patients had more frequent dental visits--every 3-4 months (40%)--than controls (19%). In parallel, 78% of the PSS patients brushed their teeth more than twice daily, compared with 28% of the control group. The PSS patient group reported having had more teeth extracted, more trouble with their teeth during lifetime, and higher expenses for dental treatment than controls. In spite of the more regular oral health care practices than the general population, PSS patients had experienced more dental caries and more radical dental treatment. It is suggested that the National Health Insurance should give emphasis to preventive care to patients with PSS. PMID- 11501879 TI - Innate and acquired immunity, cytokines, and genetic factors in relation to the mucosal immune system. PMID- 11501880 TI - Toll-like receptors and their signaling mechanism in innate immunity. AB - In Drosophila the Toll family, a group of transmembrane proteins, plays crucial roles in the host defense against invading pathogens. Mammalian species also conserve this system as the Toll-like receptor (TLR) family, which includes more than 10 members that have been identified so far. Both the Toll and TLR families recognize various kinds of microorganisms through pathogen-associated molecular patterns. Mammalian TLRs are expressed on macrophages and dendritic cells and mediate the signal for cytokine release or upregulation of costimulatory molecules. These activities cooperatively generate host defense mechanisms. Recently, gene targeting experiments, including ours, have contributed much to clarifying not only the function but also the signaling mechanism of TLRs. TLR2 is essential for recognizing lipopeptides and lipoproteins from several microorganisms and also peptidoglycans derived from gram-positive bacteria. TLR4 recognizes lipopolysaccharides and lipoteichoic acids from gram-negative and- positive bacteria, respectively. Furthermore, TLR9 is critical for recognizing bacterial DNAs. Thus, TLRs distinguish various immunostimulatory molecular patterns. Although TLR9 can produce similar biological responses, studies with mutant mice lacking a TLR-associating protein, MyD88, showed that TLR signaling is differentially regulated among TLR family members. Here, we describe recent progress in elucidating the function and signaling mechanisms of the TLR family. PMID- 11501881 TI - Porphyromonas gingivalis lipopolysaccharide: an unusual pattern recognition receptor ligand for the innate host defense system. AB - Lipopolysaccharide (LPS) is a key inflammatory mediator. Due to its ability to potently activate host inflammatory and innate defense responses, it has been proposed to function as an important molecule that alerts the host of potential bacterial infection. However, although highly conserved, LPS contains important structural differences among different bacterial species that can significantly alter host responses. For example, LPS obtained from Porphyromonas gingivalis, an etiologic agent for periodontitis, causes a highly unusual host innate host response. It is an agonist for human monocytes and an antagonist for human endothelial cells. Correspondingly, although it activates p38 MAP kinase in human monocytes, P. gingivalis LPS does not activate p38 nor ERK MAP kinase in endothelial cells. In fact, P. gingivalis LPS is an effective inhibitor of Escherichia coli LPS induced p38 phosphorylation. These data show that P. gingivalis LPS modulates host defenses in endothelial cells by interfering with MAP kinase activation. In addition, P. gingivalis LPS is unusual in that it engages TLR-2 but not TLR-4 when examined in stably transfected CHO cell lines. We propose that, since LPS is a key ligand for the human innate host defense system, these unusual properties of P. gingivalis LPS are associated with the bacterium's role in the pathogenesis of periodontitis. PMID- 11501882 TI - The multiple roles of major histocompatibility complex class-I-like molecules in mucosal immune function. AB - The human major histocompatibility complex (MHC) on chromosome 6 encodes three classical class-I genes: human leukocyte antigens (HLA) A, B, and C. These polymorphic genes encode a 43- to 45-kDa cell surface glycoprotein that, in association with the 12-kDa beta2-microglobulin molecule, functions in the presentation of nine amino acid peptides to the T-cell receptor of CD8-bearing T lymphocytes and killer inhibitory receptors on natural killer cells. In addition to these ubiquitously expressed, polymorphic proteins, the human genome also encodes several nonclassical MHC class-I-like, or class Ib, genes that, in general, encode nonpolymorphic molecules involved in various specific immunological functions. Many of these genes, including CD1, the neonatal Fc receptor for IgG, HLA-G, HLA-E, the MHC class-I chain-related gene A, and Hfe, are prominently displayed on epithelial cells, suggesting an important role in epithelial cell biology. PMID- 11501884 TI - Clinical, pathological and immunological aspects of periodontal disease. AB - The inflammatory and immune responses during the development and progression of periodontitis are reviewed. Susceptibility to periodontitis may be related to whether plasma cells predominate in the tissues of an individual, or a site, in response to the microbial insult from dental plaque. The tendency for an individual or site to form an extensive plasma cell infiltrate may indicate an inability to defend against periodontopathogenic bacteria and thus a predisposition to periodontitis. Selected pertinent areas of current interest in cellular and humoral immunology are considered within the periodontal context. These topical issues include (a) homing of immune and inflammatory cells to target tissues; (b) the local proliferation and synthetic activity of immune and inflammatory cells; (c) the cytokine profile of the inflammatory and immune cells; and (d) the immunoglobulin subclasses of locally produced antibodies. PMID- 11501883 TI - The mucosal immune system: from specialized immune defense to inflammation and allergy. AB - The mucosal immune system is a first line of defense against foreign antigens, including microbial and dietary antigens. Under normal circumstances, the mucosal immune system employs tightly regulated dynamic mucosal intra- and internets consisting of inductive and effector sites for the induction of an appropriate immunological homeostasis between the host and mucosal environments. The common mucosal immune system (CMIS), which interconnects between inductive (e.g. Peyer patch) and effector (e.g. intestinal lamina propria) tissues for the induction of the IgA response, is well characterized. Recent results provide strong evidence for the presence of a CMIS-independent IgA induction pathway. Two distinct subsets of mucosal IgA-committed B cells termed B-1 and B-2, are associated with CMIS-independence and CMIS-dependent cascades respectively. In some cases, the breakdown of this tightly regulated mucosal immune system leads to pathological responses to different gut environmental antigens. As a result, disorders such as inflammatory bowel disease (e.g. IBD) and allergic gastroenteropathy can be evoked in the gastrointestinal tissues. Recently, many studies have described possible molecular and cellular mechanisms for this dysfunction in the gastrointestinal tissues by using murine models with specific gene manipulation. In this review we summarize recent findings from our group concerning the CMIS dependent and CMIS-independent IgA induction pathways and gastrointestinal diseases (IBD and intestinal allergic diseases). These observations may provide useful information for the development of new mucosal immune therapy. PMID- 11501885 TI - Periodontal diseases: to protect or not to protect is the question? AB - For decades, investigations have identified local and systemic humoral immune responses to microorganisms comprising the supra- and subgingival biofilms in the oral cavity. Inflammation and tissue destruction in the periodontium are accompanied by alterations in the quantity, quality, and specificity of antibody. The conundrum in this scenario is the existence of a substantial plasma cell infiltrate at sites of periodontal lesions and a seemingly robust antibody response in the oral cavity and the serum, apparently coincident with progressing disease. Consequently, much effort has been expended to elucidate the critical characteristics of protective humoral responses and to develop strategies for enhancing these unique features. We and others have conducted studies attempting to distinguish disease susceptibility associated with: i) variations in response levels significantly increased to some species with disease, minimal response to others; ii) functional comparisons of antibody subclass differences, genetic regulation, and maturation of responses; iii) microbial and antigenic specificity of the antibody focus on specific pathogens and identification of selected antigens as targets for immunoprotection; and, iv) kinetics of responses during disease and therapeutic interventions linking immune changes with infection and as a measure of treatment success. This report summarizes varied research designs and results, to provide a profile of antibody in health, gingivitis, and periodontitis. These profiles may be used to provide a framework focusing on the humoral response to commensal microorganisms and likely pathogens, as they emerge in the biofilm--etiologic for or in response to disease processes. Models for antibody as a diagnostic adjunct and for predicting protective antibody responses are suggested. These concepts are likely relevant for considering vaccine approaches to periodontitis. PMID- 11501886 TI - Cytokines in periodontal disease: where to from here? AB - Numerous studies have attempted to elucidate the cytokine networks involved in chronic periodontitis, often with conflicting results. A variety of techniques were used to study cells in situ, cells extracted from gingival tissues, peripheral blood mononuclear cells, purified cell populations, and T cell lines and clones. Bacterial components, including sonicates, killed cells, outer membrane components, and purified antigens, have all been used to stimulate cells in vitro, making comparisons of cytokine profiles difficult. As it is likely that different cells are present at different disease stages, the inability to determine disease activity clinically is a major limitation of all these studies. In the Context of tissue destruction, cytokines such as IL-1, IL-6 and IL-18 are likely to be important, as are their regulating cytokines IL-10 and IL-11. In terms of the nature of the inflammatory infiltrate, two apparently conflicting hypotheses have emerged: one based on direct observations of human lesions, the other based on animal experimentation and the inability to demonstrate IL-4 mRNA in gingival extracts. In the first of these, Th1 responses are responsible for the stable lesion, while in the second Th2 responses are considered protective. Using Porphyromonas gingivalis-specific T cell lines we have shown a tendency for IFN-gamma production rather than IL-4 or IL-10 when antigen is presented with peripheral blood mononuclear cells which may contain dendritic cells. It is likely that the nature of the antigen-presenting cell is fundamental in determining the nature of the cytokine profile, which may in turn open up possibilities for new therapeutic modalities. PMID- 11501887 TI - Immune mechanisms in oral lichen planus. AB - Although we still don't know the cause, there has been much research into the immune and pathological mechanisms that underlie oral lichen planus (OLP) and it is now possible to piece together a much clearer picture of the disease process. There is consensus that in OLP there is chronic, cell-mediated, immune damage to basal keratinocytes in the oral mucosa that are recognized as being antigenically foreign or altered. In most cases, however, the identity of the target antigen remains unknown. It is likely that cytokines released by the affected keratinocytes, and the associated inflammatory infiltrate, play a key role in the selective recruitment of the T-cell-dominated infiltrate that characterizes OLP, through their ability to induce adhesion molecule expression as well as further cytokine and chemokine release. In susceptible individuals, chronic presentation of antigen by basal keratinocytes may perpetuate the condition and direct cell mediated immune damage on the keratinocytes. PMID- 11501888 TI - Cytokine network and dysregulated apoptosis in atopic dermatitis. AB - Activation and skin-selective homing of peripheral blood memory/effector T cells and effector functions in the skin represent sequential immunological events in the pathogenesis of atopic dermatitis (AD). T cells infiltrating the skin utilize the cutaneous lymphocyte-associated antigen (CLA) and other receptors to recognize and cross the vascular endothelium. In the peripheral blood of AD patients, both CD4+ and CD8 subsets of CLA+CD45RO+ T cells are in an activated state with high CD25, HLA-DR, and CD40-ligand expression. They express upregulated Fas and Fas-ligand and undergo activation-induced apoptosis. After homing to skin these T cells form dermal infiltrates which play a key role in the pathogenesis of the disease. Skin-infiltrating T cells in AD are protected from activation-induced cell death, although they express both Fas and Fas-ligand. They are protected from apoptosis by cytokines such as IL-2, IL-4, and IL-15 and extracellular matrix components such as fibronectin and transferrin. CLA+, skin homing T cells may play a role in peripheral blood eosinophilia and hyper IgE production by high IL-5 and IL-13 expression, respectively. These T cells secrete IFN-gamma in the skin, which upregulates Fas on keratinocytes and renders them susceptible to apoptosis. Keratinocyte apoptosis is induced by Fas-ligand, either soluble or expressed on the surface of T cells, leading to eczema formation. Here we discuss the mechanisms of skin-selective T cell homing and activation, and emphasize the concept of dysregulated apoptosis of T cells, eosinophils, and keratinocytes as essential pathogenetic episodes in AD and other eczematous disorders. PMID- 11501889 TI - Genes and environment in celiac disease. AB - Celiac disease is an intestinal disorder that develops as a result of interplay between genetic and environmental factors. HLA genes along with non-HLA genes predispose to the disease. Linkage studies have failed to identify chromosomal regions other than the HLA region which have major effects, indicating the existence of multiple non-HLA predisposing genes with modest effects. Association studies have shown that CTLA4 or a closely located gene is one of these genes. The primary HLA association in the majority of celiac disease patients is with DQ2 (DQA1*05/DQB1*02) and in the minority of patients with DQ8 (DQA1*0301/DQB1*0302). Gluten reactive CD4+ T cells can be isolated from small intestinal biopsies of celiac patients but not from controls. DQ2 or DQ8, but not other HLA molecules carried by patients, present peptides to these T cells. A number of distinct T cell gluten epitopes exist, most of them posttranslationally modified by deamidation. DQ2 and DQ8 bind the epitopes such that the glutamic acid residues created by deamidation are accommodated in pockets that have a preference for negatively charged side chains. There is evidence that deamidation in vivo is mediated by the enzyme tissue transglutaminase (tTG). Overall, the results point to control of the immune response to gluten by intestinal T cells restricted by the DQ2 or DQ8 molecules. This is likely to be a critical checkpoint for the development of celiac disease and could explain the dominant genetic role of HLA in this disorder. The products of the other predisposing genes may participate in pathway(s) that lead(s) to lesion formation. The minor genetic effects of the non-HLA genes could indicate a lack of critical checkpoints along these pathways, or that there are several pathways leading to the lesion formation. PMID- 11501890 TI - Genetics of inflammatory bowel disease: from bench to bedside? AB - Epidemiological data, particularly concordance rates in twin pairs and in multiply affected families, provide strong evidence that both genetic and environmental influences are important in the development of the chronic intestinal inflammation characteristic of Crohn disease and ulcerative colitis. Furthermore, supplementary data now suggest that not only susceptibility, but also disease behavior and response to therapy may have a strong genetic influence. The model of disease susceptibility most pertinent to the inflammatory bowel diseases is that Crohn disease and ulcerative colitis are related polygenic disorders. Recently, this model has received strong support from the results of genome-wide scanning and candidate gene studies carried out in European, North American and Australian populations. In spite of all potential difficulties related to disease and ethnic heterogeneity, consistent replication of linkage has been found with distinct regions on chromosomes 16, 12, 6 (the major histocompatibility complex) and most recently chromosome 14. Whereas the linkages on chromosome 16 and 14 appear strongest in Crohn disease, the chromosome 12 locus appears most pertinent to ulcerative colitis, and the HLA region appears more pertinent in all forms of inflammatory bowel disease. The current challenge is to narrow these regions of linkage and identify the susceptibility genes within the regions. The task may be greatly benefited by the recent successful sequence data available from the human genome project. Compelling data have emerged to suggest genetic markers that may allow prediction of disease severity, and efficacy and tolerability of immuno-suppressants commonly used in inflammatory bowel disease. PMID- 11501891 TI - Effects of providing and withholding postfitting fine-tuning adjustments on outcome measures in novice hearing aid users: a pilot study. AB - This pilot study was designed to examine the effects of either providing or withholding subject-driven, postfitting electroacoustic changes (fine tuning) on hearing aid performance as measured by tests of speech recognition in noise, sound quality, and benefit. A group of adult first-time hearing aid users with moderate, high-frequency sensorineural hearing loss was studied over a 5-month postfitting period. Half of the group served as a control group; half constituted the treatment group. Treatment consisted of making as many postfitting electroacoustic manipulations as the participants' complaints/observations required. These adjustments were withheld from the control group individuals. Group performance differences were assessed using the COSI, two versions of the APHAB, a Satisfaction scale, a Sound Quality tool, and the SIN test. Analysis revealed no statistically significant differences in the control and treatment groups' performances or perceived benefit at any of the five postfitting test sessions. These data suggested that there was no measurable advantage to be derived from making postfitting frequency-gain adjustments of 10 dB or less to hearing aids with wide dynamic range compression fitted in the manner described in this article. Definitive studies of the effects of postfitting electroacoustic fine tuning must be undertaken. PMID- 11501892 TI - The honor of the "invitation" to participate in the editorial review process: giving a little bit back. PMID- 11501893 TI - Parents' reactions and recommendations after diagnosis and hearing aid fitting. AB - This study was designed to explore parent reactions to the early stages of audiologic assessment and intervention. A total of 213 parents whose children were under the age of 6 years returned a mail survey. Respondents from 45 states participated. Parents were asked to (1) report the approximate age of diagnosis and hearing aid fitting; (2) comment on reasons for any delays encountered from diagnosis to fitting; and (3) respond to questions concerning their reactions to the initial fitting of amplification. The median age of identification was earlier than some previous investigations; however, substantial delays occurred between diagnosis and hearing aid fitting. Reasons for delay included the need for further audiologic evaluation, problems obtaining return appointments, illness of the child, and difficulties obtaining adequate earmolds. Parent reactions to hearing aids, once fitted, included concerns about appearance and questions about maintenance and use, but attitudes regarding hearing aids and their perceived benefits improved over time. PMID- 11501894 TI - Universal neonatal hearing screening: past, present, and future. AB - After a brief review of the history of newborn hearing screening including the Downs behavioral testing procedure, the Crib-o-gram and similar devices, and the use of auropalpebral reflex and otoacoustic emissions, there is a discussion of key issues that need to be resolved before universal hearing screening is introduced. Included are questions regarding the target population(s) of screening programs, well baby versus NICU screening, dealing with false-positives and the effects on parent-child relationships, and finally, the availability of resources for screening and follow-up. The results of a recent study in the United Kingdom that assessed the current state of audiology services and found there is a difference between existing standards and what is actually being done in practice, are presented and considered in terms of current trends in the United States to move ahead with universal screening without a solid database of information regarding the preparedness of clinical centers to deal with the need for services that will result from the initiation of universal programs. Caution is urged. PMID- 11501895 TI - Survey of hearing aid fitting practices for children with multiple impairments. AB - The fitting of amplification on young children with multiple impairments in addition to hearing loss is a challenge faced regularly by audiologists. However, very little has been published on this topic in the audiological literature. The purpose of this survey was to document hearing aid fitting practices for this population within the United States. Specifically, audiologists who regularly serve children were asked to complete a series of questions on their educational preparation and their hearing aid selection, fitting, and verification practices for children with multiple impairments. For purposes of this survey, multiple impairments included vision impairment, mental retardation, physical impairment, and autism spectrum disorders. Findings from this survey suggest that children with special needs in addition to hearing loss are typically fit in the same way and with the same type of amplification as those with hearing loss only. In addition, differences were noted in hearing aid selection, fitting, and verification practices across work settings. Future directions and research needs are suggested. PMID- 11501896 TI - Practical considerations of real-ear-to-coupler difference measures in infants. AB - The purpose of this study was to investigate some of the practical issues surrounding the use of the real-ear-to-coupler difference (RECD) measure in infants in clinical settings. Twenty-two newborns were seen once a month during their first year of life for the purpose of obtaining RECD measurements. Two methods for obtaining the RECD were examined: the commonly used constant insertion depth method and the acoustic method. Differences between the results of the two methods were examined as well as test-retest reliability for each. Findings suggest that no compelling evidence exists to support the use of one of these methods over the other. Use of either method resulted in good test-retest reliability in the midfrequency range of the RECD, with poorer reliability on both ends of the frequency spectrum. Test-retest reliability decreased with increasing age for both methods. In addition, reliability of between-ear RECD values was examined. Between-ear measurements were most reliable in the midfrequency range, and reliability decreased slightly with increasing age. PMID- 11501897 TI - Determination of azadirachtin in agricultural matrixes and commercial formulations by enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for azadirachtin (aza), a biopesticide from the neem tree (Azadirachta indica A. Juss). The immunogen was synthesized by epoxidation using the furan ring in the aza molecule. Rabbits were immunized with either bovine serum albumin (BSA) azadirachtin or ovalbumin (OA)-azadirachtin conjugate. Evaluation of the antisera by antibody capture assay showed that the antibody titer of antisera raised against OA-aza was 1:30,000. An indirect competitive ELISA was developed with BSA azadirachtin as coating antigen and aza-specific antibodies raised against OA-aza immunogen. The immunoassay showed an inhibitory concentration (IC50) value of 75 ppb, with a range of detection from 0.5 to 1,000 ppb for azadirachtin [based on regression analysis, y= 85.87 (-18.89x); r2 = -0.97]. Cross-reactivity of the antibodies with 2 aza- derivatives (22,23-dihydro-23beta-methoxy azadirachtin and 3-tigloylazadirachtol) was 33 and 29%, respectively. The indirect competitive ELISA was validated and evaluated by quantitating aza in spiked agricultural commodities and from neem formulations. Azadirachtin was spiked into 5 different agricultural commodities: tomato, brinjal, coffee, tea, and cotton seed at 500 and 1,000 ppb and recovered at 62-100%. In samples drawn from 6 lots, the aza content in neem-seed kernels ranged from 0.1 to 0.15%; in commercial neem formulations the content ranged from 200 to 2,000 ppm. The method developed may be applied to environmental monitoring of aza and quality assurance studies of aza-based commercial formulations. PMID- 11501898 TI - Determination of manganese in feedstuffs by flow injection spectrophotometry based on rapid formation of permanganate at room temperature. AB - A simple and rapid flow injection spectrophotometric procedure was developed for determination of manganese. In the presence of pyrophosphate and acetate, manganese was immediately oxidized to permanganate by periodate at room temperature in slightly alkaline medium. Under optimized conditions, the determination was made with a sampling rate of 120/h, a linear range of 0-30 mg/L Mn(II), a detection limit (S/N = 3) of 0.08 mg/L, and a relative standard deviation of 0.6% (n = 11) at 10 mg/L Mn(II). The proposed method was used to determine manganese in trace mineral premixes and feedstuffs. Results agreed well with those obtained by the standard atomic absorption spectroscopy method. PMID- 11501899 TI - Spectrofluorimetric determination of vigabatrin and gabapentin in dosage forms and spiked plasma samples through derivatization with 4-chloro-7-nitrobenzo-2-oxa 1,3-diazole. AB - A highly sensitive and specific method is proposed for the determination of vigabatrin (I) and gabapentin (II) in their dosage forms and spiked human plasma. The method is based on coupling the drugs with 4-chloro-7-nitrobenzo-2-oxa-1,3 diazole in borate buffer at pH 7.1 and measuring the resulting fluorescence at 532 nm after excitation at 465 nm. The fluorescence intensity was a linear function of the concentration of the drugs over the ranges of 1.3-6.5 and 1.7-8.5 microg/mL for I and II, respectively. Minimum detectability values were 0.54 microg/mL (4.2 x 10(-6)M) and 0.97 microg/mL (5.7 x 10(-6)M) for I and II, respectively, under the described conditions. The proposed method was successfully applied to the determination of the 2 drugs in their dosage forms, and the percent recoveries +/- standard deviation (SD) were 104.53 +/- 1.2 and 100.00 +/- 1.32 of the label claim for I and II, respectively. The method was further applied to the determination of vigabatrin in spiked plasma samples. The percent recovery +/- SD was 101.58 +/- 2.68. Interference from endogenous alpha amino acids was overcome through selective complexation with freshly prepared Cu(OH)2. The interference likely to be encountered from co-administered drugs, such as carbamazepine, cimetidine, clonazepam, clopazam, phenobarbital, valproic acid, and lamotrigine, was also studied. A reaction pathway is suggested. PMID- 11501900 TI - Determination of clenbuterol residues in bovine urine by optical immunobiosensor assay. AB - Clenbuterol (CBL) is an orally active beta2-adrenoceptor agonist which has been used in veterinary medicine as a broncodilator and an agent of uterine relaxation. It has however become better known as a drug used illegally to promote growth in farm animals. A rapid and sensitive biosensor assay was developed to detect CBL residues in bovine urine. The method involved a simple extraction procedure using tert-butyl methyl ether followed by analysis on the biosensor with results obtained against a buffer calibration curve. The assay allowed up to 88 samples to be analyzed per working day, with each cycle on the biosensor taking approximately 7 min to complete. The limit of detection (LOD) was determined as 0.27 ng/mL using 20 EU reference blank urine samples. The intra assay Sr ranged from 4.7-7.6% for 3 control samples while the interassay Sr ranged from 9.2-12.7%. The recovery was found to be approximately 95%. A series of incurred urine samples were assayed and the results compared by Enzyme immunoassay (EIA), radio-immunoassay (RIA), and gas chromatography/mass spectrometry (GC/MS) analysis. Urine samples taken from local abattoirs were also analyzed by the biosensor method and by EIA analysis. The antibody used in the biosensor test exhibited high cross reactivity with at least 7 other beta agonists allowing detection of these compounds at less than 1 ng/mL in bovine urine. PMID- 11501901 TI - Determination of pyrimethamine in animal tissue and egg by liquid chromatography with fluorescence detection. AB - A sensitive and selective liquid chromatographic (LC) assay was developed to determine the concentration of pyrimethamine in animal tissue and egg by fluorescent derivative. Animal samples were extracted with acetonitrile, centrifuged, and purified by hexane. Fluorescent derivatization was performed by reacting pyrimethamine with chloroacetaldehyde and subjected to LC with fluorescence detection (excitation wavelength 300 nm, emission wavelength 420 nm). The limit of detection was 10 ng/g (10 ppb) and the standard calibration curve was linear in the range of 1-100 ppb (0.01-1 ng/10 microL). Recoveries from samples fortified at levels of 0.1 and 1 ppm (microg/g) were 61.0-77.4 and 65.5 81.2%, respectively. The method was applied to the monitoring of marketed samples. Pyrimethamine was not determined in any of the 70 samples: 20 swine muscle; 20 chicken muscle; 10 chicken liver; and 20 egg. PMID- 11501902 TI - Comparison of liquid chromatography/mass spectrometry, ELISA, and phosphatase assay for the determination of microcystins in blue-green algae products. AB - More than 100 samples of blue-green algae products (consisting of Aphanizomenon, Spirulina, and unidentified blue-green algae) in the form of pills, capsules, and powders were collected from retail outlets from across Canada. The samples were extracted with 75% methanol in water and centrifuged to remove solids. Aliquots of the extracts along with spiked blank sample extracts were sent to each participating laboratory and independently analyzed for microcystins by enzyme linked immunosorbent assay (ELISA), protein phosphatase inhibition assay, and by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after sample cleanup using C18 solid-phase extraction. The results obtained by ELISA and LC-MS/MS agreed very well over a concentration range of about 0.5-35 microg/g. The colorimetric phosphatase results generally agreed with the other 2 methods. While the 2 biochemical assays measured total microcystin content compared with a standard of microcystin LR, the LC-MS/MS method measured specific microcystins (LA, LR, RR, YR) using external standards of these for identification and quantitation. Microcystin LR was found in all positive samples by LC-MS/MS. Microcystin LA was the only other microcystin found in the samples analyzed. These 2 microcystins represent essentially all the microcystins that were present in the extracts. Otherwise, the LC-MS/MS results would have been significantly lower than the results of the biochemical assays had other unknown microcystins been present. PMID- 11501903 TI - Simultaneous determination of magnesium L-ascorbyl-2-phosphate and kojic acid in cosmetic bleaching products by using a microbore column and ion-pair liquid chromatography. AB - A rapid, sensitive, and reliable method was developed for the simultaneous determination of magnesium L-ascorbyl-2-phosphate (AS-PM) and kojic acid (KA), commonly using bleaching agents in cosmetic products. The method uses a prepacked C18 microbore column for ion-pair liquid chromatography with photodiode array detection. The eluant is 0.5mM tetrabutylammonium bromide and 0.05M phosphate buffer, pH 5, containing 5% methanol, at a flow rate of 70 microL/min. The determinations of AS-PM and KA were linear in the range of 0.5-1,000 and 0.25-500 ng, respectively. The precision of the assay ranged from 0.31 to 2.19%, and the detection limits (signal-to-noise ratio = 3) were 0.04 and 0.02 ng for AS-PM and KA, respectively. PMID- 11501904 TI - Liquid chromatographic determination of nystatin in pharmaceutical preparations. AB - A rapid, reversed-phase liquid chromatographic method was developed for the assay of nystatin in the bulk drug and a variety of dosage forms. Analysis was performed on a Symmetry C18 reversed-phase column using a mobile phase of methanol-water-dimethylformamide (DMF; 55 + 30 + 15, v/v/v), with detection by UV at 305 nm. Quantitation is based on the sum of the peak areas of the 2 major isomers of nystatin. The linearity of the assay was determined for a concentration range of 0.05 to 0.2 mg/mL (correlation coefficient > 0.999). Accuracies and precision showed good reproducibility. PMID- 11501905 TI - Determination of organic acids of low molecular weight and phosphate in soil by capillary electrophoresis. AB - Capillary zone electrophoresis with indirect UV detection at 254 nm was found to be suitable for the determination of organic acids and phosphate in aqueous extracts of soil. The best support electrolyte solution was found to be 10 mM p hydroxybenzoic acid with 0.5 mM tetradecyltrimethylammonium bromide to reverse electroosmotic flow. This methodology was tested with 9 analytes found in soils: acetate, citrate, formate, phosphate, lactate, oxalate, pyruvate, succinate, and tartrate. The results obtained show that the methodology is adequate for most of the analytes. The sensitivity to oxalate and citrate was low, and the high concentrations of major inorganic anions interfered with the detection of the former. The methodology was applied to the analysis of aqueous extracts of soil samples. Formate, phosphate, lactate, and acetate anions were detected in most of the samples. PMID- 11501906 TI - Spectrophotometric determination of sulfur dioxide in air, using thymol blue. AB - A simple and sensitive spectrophotometric method was developed for the determination of trace amounts of sulfur dioxide. The method is based on the reaction of SO2 with a known excess of ICI as the oxidant. The unreacted ICI iodinates thymol blue under acidic conditions. The lambdamax of thymol blue is at 545 nm under acidic conditions, and on lodination lambdamax shifts to 430 nm. This shift results in a decrease in the absorbance at 545 nm. The amount of uniodinated thymol blue present depends on the concentration of unreacted ICI, which in turn depends on the SO2 concentration. The system obeys Beer's law in the range 0-30 microg SO2 in a final volume of 25 mL, having a molar absorptivity of 3.2 x 10(4) L/mol cm with a relative standard deviation (RSD) of 2% at 24 microg SO2 (n = 10). The uniodinated dye can be extracted into 5 mL isoamyl alcohol under acidic conditions for measurement of absorbance. The extraction method obeys Beer's law in the range 0-5 microg SO2, having a molar absorpitivity of 4.16 x 10(4) L/mol x cm with an RSD of 1.9% at 4 microg SO2 (n = 10). The method has been successfully applied to the determination of atmospheric SO2. PMID- 11501907 TI - Stability and recovery of triazine and chloroacetamide herbicides from pH adjusted water samples by using empore solid-phase extraction disks and gas chromatography with ion trap mass spectrometry. AB - Empore disks were used to successfully extract herbicide residues from a difficult-to-analyze surface water source and deionized water. Herbicide recoveries were lower in surface water at 7,14, or 21 days after fortification and storage at 4 degrees C, presumably due to chemical sorption onto precipitated organic particulates. The addition of acid to the samples, as recommended in EPA Method 525.2, did not affect recoveries of alachlor and metolachlor, but reduced recoveries of atrazine, simazine, and cyanazine. Treatment of water samples with sodium hypochlorite did not affect alachlor or metolachlor recoveries, but greatly reduced the recovery of all triazine herbicides. This indicates that addition of acid or sodium hypochlorite to water samples may be detrimental to triazine analysis. PMID- 11501908 TI - Determination of phenolic xenoestrogens in environmental samples by liquid chromatography with mass spectrometric detection. AB - A method is proposed for the determination of several phenolic xenoestrogens in aqueous and solid environmental samples. The method uses solid-phase extraction (preceded by ultrasonic solvent extraction for solid samples), reversed-phase liquid chromatographic separation, and mass spectrometric detection using both atmospheric pressure chemical ionization and electrospray ionization. This method was developed to support several studies undertaken to obtain aquatic and sedimentary data for rivers and seashores in Spain that are likely to be contaminated by endocrine-disrupting compounds (EDCs) as a consequence of wastewater discharge. Nonylphenol polyethoxylates (NPEOs), nonylphenoxy carboxylates (NPECs), nonylphenol (NP), octylphenol (OP), and bisphenol A (BPA) were determined in various samples of surface water and sediment, collected at different locations upstream and downstream from outfalls of municipal wastewater treatment plants (WWTPs). Seawater and marine sediments were collected in different harbor areas in Spain. Additionally, WWTP influent and effluents were analyzed to monitor the occurrence and transformation of phenolic EDCs during physicochemical and biological treatment. Rather high concentrations of the compounds investigated were found in some samples. Concentrations of NP were < or = 590 microg/kg in sediments and < or = 15 microg/L in water samples. NPEOs and NPECs were found in water samples in concentrations < or = 41 and < or = 35 microg/L, respectively. In solid samples (river sediment), concentrations of NPEO were < or = 818 microg/kg and those of NP1EC were 95 microg/kg. PMID- 11501909 TI - Sensitivity and specificity of the BAX for screening/Listeria monocytogenes assay: internal validation and independent laboratory study. AB - In this study to certify the BAX for Screening/Listeria monocytogenes assay (DuPont Qualicon, Wilmington, DE), an internal evaluation was conducted on 16 food types that were simultaneously analyzed with the BAX system (BAX), and the ISO method for the detection of L. monocytogenes (ISO). No statistically significant difference in performance between the BAX and ISO methods was observed. Inclusivity/exclusivity testing showed that the BAX system was able to detect 97 of 97 (100%) of L. monocytogenes strains tested. None of 56 other Listeria species or non-Listeria tested gave a reproducible positive BAX result. Ruggedness testing demonstrated that performance of the assay was not affected by reasonable variability in the operating parameters. BAX was then submitted for independent laboratory validation. In this phase, BAX was compared with standard culture methods for the detection of L. monocytogenes in chicken (USDA-FSIS), crab meat (BAM), and milk (AOAC). This study validated product claims of sensitivity and specificity >98% in accordance with AOAC Performance Tested Method requirements. PMID- 11501910 TI - Quantitative determination of paralytic shellfish poisoning toxins in shellfish by using prechromatographic oxidation and liquid chromatography with fluorescence detection. AB - The prechromatographic oxidation LC method developed by Lawrence [J. Assoc. Off. Anal. Chem. 74, 404-409(1991)] for the determination of paralytic shellfish poisoning (PSP) toxins has been tested for the quantitative determination of PSP toxins in shellfish. All aspects of the method were studied and modified as necessary to improve its performance for routine regulatory purposes. The chromatographic conditions were changed to shorten analysis time. The oxidation reaction was tested for repeatability and the influence of the sample matrix on quantitation. An important part of the study was to quantitatively evaluate an ion exchange (-COOH) cleanup step using disposable solid-phase extraction cartridges that separated the PSP toxins into 3 distinct groups for quantitation, namely the C toxins, the GTX toxins, and the saxitoxin group. The cleanup step was very simple and used increasing concentrations of aqueous NaCl for elution of the toxins. The C toxins were not retained by the cartridges and thus were eluted unretained with water. The GTX toxins (GTX1 to GTX6 as well as dcGTX2 and dcGTX3) eluted from the cartridges with 0.05M NaCl while the saxitoxin group (saxitoxin, neosaxitoxin, and dcsaxitoxin) required 0.3M NaCl for elution. Each fraction was analyzed by LC after oxidation with periodate or peroxide. All of the compounds could be separated and quantitatively determined in spiked samples of mussels, clams, and oysters. The nonhydroxylated toxins could be quantitated at concentrations as low as about 0.02 microg/g (2 micro/100 g) of tissue while the hydroxylated toxins could be quantitated at concentrations as low as about 0.1 microg/g (10 microg/100 g). Average recoveries of the toxins through the complete cleanup procedure were 85% or greater for spiked extracts of oysters and clams and greater than 73% for mussels. PMID- 11501911 TI - Characterization of partially transesterified poly(beta-hydroxyalkanoate)s by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used for the characterization of a partially transesterified poly(beta-hydroxyalkanoate) (PHA), a polymer produced by the bacterial strain Alcaligenes eutrophus with saponified vegetable oils as the sole carbon sources. The transesterification was carried out separately under acidic and basic conditions to obtain PHA oligomers weighing <10 kDa. The intact oligomers were detected in their cationized forms, [M + Na]+ and [M + K]+, by MALDI-TOFMS. A composition analysis, using the MALDI-TOF spectra, indicated that the oligomers obtained via acid catalysis contained a methyl 3-hydroxybutyrate end group, and those obtained by base catalysis had a methyl crotonate (olefinic) end group. In addition to hydroxybutyrate (HB), the oligomers were found to contain a small percentage of hydroxyvalerate, which was independently confirmed by gas chromatography/mass spectrometry. In comparison, analysis of a commercial PHA polymer, transesterified under identical conditions, showed only the presence of HB, i.e., a pure poly(HB) homopolymer. PMID- 11501912 TI - Determination of aflatoxin B1 in baby food (infant formula) by immunoaffinity column cleanup liquid chromatography with postcolumn bromination: collaborative study. AB - A collaborative study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for determination of aflatoxin B, in a milk powder based infant formula at a possible future European regulatory limit (0.1 ng/g). The test portion was extracted with methanol-water (8 + 2 [v + v]), filtered, diluted with water, and applied to an immunoaffinity column. The column was washed with water to remove interfering compounds, and the purified aflatoxin B1 was eluted with methanol. The separation and determination of the aflatoxin B1 was performed by reversed-phase LC and detected by fluorescence after postcolumn derivatization (PCD) involving bromination. PCD was achieved with either pyridinum hydrobromide perbromide (PBPB) or an electrochemical (Kobra) cell by addition of bromide to the mobile phase. The baby food (infant formula) test samples, both spiked and naturally contaminated with aflatoxin B1, were sent to 14 laboratories in 13 different European countries. Test portions were spiked at levels of 0.1 and 0.2 ng/g for aflatoxin B1. Recoveries ranged from 101 to 92%. Based on results for spiked test samples (blind pairs at 2 levels) and naturally contaminated test samples (blind pairs at 3 levels), the relative standard deviation for repeatability (RSDr) ranged from 3.5 to 14%. The relative standard deviation for reproducibility (RSDR) ranged from 9 to 23%. Nine participants used PBPB derivatization, and PMID- 11501913 TI - Proposed phytic acid standard including a method for its analysis. AB - A method is described that accurately and rapidly quantifies the free and total phosphorous content of a commercially available, purified, phytic acid preparation. This allows its use as a standard for phytic acid determinations in foods. The method involves a wet ashing step followed by phosphorous measurement with a 1-amino-2-naphthol-4-sulfonic acid-molybdate reagent in a microplate reader at 660 nm. The procedure can be performed in 3 h with as little as 50 mg sample. PMID- 11501914 TI - Quantitation of volatile oils in ground cumin by supercritical fluid extraction and gas chromatography with flame ionization detection. AB - Ground cumin is used as a flavoring agent in a number of ethnic cuisines. The chemical entities, which primarily establish its characteristically pungent flavor, are found in the volatile oil of cumin. Fixed oils and carbohydrates tend to round out the harshness of the volatile oil components. However, the quantity of volatile oil is commonly the measure of the quality of this spice. For several decades, the spice industry has used a classical distillation procedure for the determination of volatile oil in cumin and other spices. However, the method is cumbersome and requires nearly 8 h to complete. Supercritical fluid extraction with capillary gas chromatography-flame ionization detection is utilized in the formulation of a rapid, accurate, and specific method for the determination of volatile oil in ground cumin. Samples are extracted in a static-dynamic mode with CO2 at 550 bar and 100 degrees C. Toluene is used as a static modifier addition. The extracted volatile oil, collected in toluene, is analyzed directly using tetradecane as the internal standard. Integration is performed as grouped peaks to include all chemical entities found in cumin volatile oil recovered from the official distillation procedure. Results from this procedure compare favorably with those obtained by the official procedure (coefficient of correlation = 0.995, 24 samples). PMID- 11501915 TI - Furanocoumarins in celery and parsnips: method and multiyear Canadian survey. AB - The natural occurrence of biologically active furanocoumarins in common vegetables is an area of increasing interest with respect to human health. In this study, an efficient, rugged, and sensitive liquid chromatographic method with ultraviolet photodiode array detection was developed for the estimation of 5 biologically active furanocoumarins (psoralen, bergapten, xanthotoxin, trioxsalen, and angelicin) in celery and parsnips. When authentic samples were spiked with a mixture of furanocoumarins at individual levels of 2 to 10 microg/g, the method produced overall recoveries of 77 and 75% of all furanocoumarins from celery and parsnips, respectively. The method was applied in 2 laboratories to a multiyear survey of more than 200 samples. Of 110 parsnips samples, 109 (99%) contained quantitatable levels of furanocoumarins. The mean level of total furanocoumarins in the positive parsnip samples was 15.1 microg/g; the maximum level detected was 145 microg/g. Of 114 celery samples, 88 (77%) contained quantitatable levels of furanocoumarins. The mean level of total furanocoumarins in the positive celery samples was 1.9 microg/g; the maximum level detected was 15.2 microg/g. Xanthotoxin and bergapten were the most commonly detected furanocoumarins in both celery (68 and 63%) and parsnips (97 and 96%). Xanthotoxin had the highest mean level of positives in both celery (1.3 microg/g) and parsnips (8.5 microg/g). Little year-to-year variation in either total furanocoumarin levels or incidence was noted. PMID- 11501916 TI - Rapid determination of total trans fat content--an attenuated total reflection infrared spectroscopy international collaborative study. AB - Interest in trans fat labeling has prompted efforts to develop new, more efficient methods for rapidly and accurately determining trans fat content of foods. A novel and rapid (5 min) attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopic procedure was recently developed and applied to food products. This procedure was voted official method AOCS Cd 14d-99 by the American Oil Chemists' Society in 1999 after testing in a 12 laboratory international collaborative study. The results of the study are described in this paper. Analytical ATR-FTIR results exhibited high accuracy in the range 5-40% trans; results tended to have <2% high bias relative to the gravimetrically determined values. The precision of this internal reflection method was found to be superior to the precision of transmission infrared official methods. It is recommended that the applicability of the ATR-FTIR method be limited to trans levels of >5% (as percent of total fat). PMID- 11501917 TI - Polyvinyl chloride-based membranes for flow injection analysis of quinine in beverages. AB - Potentiometric response characteristics were evaluated for quinine selective sensors based on a lipophilic ion-exchanger potassium tetrakis[3,5 bis(trifluoromethylphenyl)]borate (PTFB) immobilized together with plasticizing solvents in polyvinyl chloride membranes. The use of dioctyl phthalate (DOP), 2 nitrophenyl phenyl ether (NPPE), and bis(2-ethylhexyl)adipate (BEHA) plasticizers produced good quality quinine sensors that were sensitive and fast responding, and exhibited near Nernstian responses when used as batch-sensors. These membranes were further tested in a wall-jet flow-through potentiometric flow injection analysis (FIA) detector. Quinine sensors containing BEHA were the most suitable membrane, with no noticeable differences in sensitivity even after 5 h of continuous exposure to solutions. Interference by foreign species such as alkali, alkaline earth metal ions, sugars, and sodium benzoate was minimal in either the batch-mode (log selectivity coefficients <-2.60) or as FIA detector. When the FIA method was used, recoveries averaged 99.70% from mineral water samples spiked with 16.22 and 162.20 ppm quinine. Results for determination of quinine in other drinks such as carbonated (64.88 and 71.37 ppm) and bitter lemon drinks (55.15 ppm) agreed with those of the manufacturer's claimed values of <68, <81, and 51 ppm, respectively. With a sample throughput of 50 samples/h under optimized flow conditions, the method can be routinely used to monitor quinine in such samples. PMID- 11501918 TI - Culture age and drying time as variables of the AOAC Sporicidal Test. AB - In the United States, the AOAC Sporicidal Activity of Disinfectants Method 966.04 is the standard for identifying a liquid chemical germicide as a sterilant. Furthermore, the highest level of a disinfectant must also be a sterilant as defined by Method 966.04, when used in its sterilant mode for a longer exposure time. The AOAC Sporicidal Test is also used as a part of the standard test methods to define a sterilant for Australia and the European Union. Many laboratories have identified variables of this test that can affect the sterilization exposure time for sterilants, or even the ability to classify a chemical as a sterilant. Method 966.04 requires spore-labeled porcelain penicylinders (cylinders) and silk suture loops, collectively referred to as carriers, to be dried for 24 h, but allows these carriers to be used for at least 7 days, in effect allowing a drying time of 24 h to at least 7 days. We tested the resistance of cylinders that had been labeled with Bacillus subtilis spores cultured for 72, 96, and 120 h, and dried for 24, 48, and 72 h against a 60 min exposure to 2.0% alkaline glutaraldehyde, and 2, 5, 10, 15, and 20 min exposures to 2.5N HCl. All the culture incubation and drying times met the standard of resistance to 2.5N HCI for at least 2.0 min at 20 degrees C, and all carriers contained at least 10(5) colony-forming units (CFU) of B. subtilis per carrier. However, for 3 repeated tests, regardless of incubation time, an average of 96% of the carriers were sterilized by the 2.0% glutaraldehyde after drying for 24 h, and an average of 61 % were sterilized after drying for 48 or 72 h. We propose that the variable of drying time be eliminated from Method 966.04. PMID- 11501919 TI - Multiresidue determination of pesticides in honey by matrix solid-phase dispersion and gas chromatography with electron-capture detection. AB - A multiresidue method was developed for the determination of 15 pesticides (organochlorines, organophosphorus compounds, pyrethroids, and other acaricides) in various commercial honeys (eucalyptus, lavender, orange, rosemary, and multifloral). The analytical procedure is based on the matrix solid-phase dispersion of honey in a mixture of Florisil and anhydrous sodium sulfate; the mixture is placed in small plastic columns and extracted with hexane-ethyl acetate (90 + 10, v/v). The pesticide residues are determined by capillary gas chromatography with electron-capture detection. Recoveries with the method at concentrations between 0.15 and 1.5 microg/g ranged from 80 to 113%, and relative standard deviations were <10% for all the pesticides studied. The pesticide detection limits were within the range 0.5-5 microg/kg for organochlorines, around 3 microg/kg for the chlorinated organophosphorus pesticides studied, near 15 microg/kg for fluvalinate, and about 3 microg/kg for the other pyrethroids. PMID- 11501920 TI - Determination of clethodim and its oxidation metabolites in crops by liquid chromatography with confirmation by LC/MS. AB - A method was developed for determination of the herbicide clethodim (C0) and its oxidation metabolites clethodim sulfoxide (C1) and clethodim sulfone (C2) in agricultural products. Upon extraction, both C0 and C1 were oxidized to C2 by m chloroperoxybenzoic acid, and C2 was determined by liquid chromatography (LC). The C2 peak was confirmed by liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization (ESI). Recoveries of C0 from radish, tomato, onion, sweet potato, kidney bean, carrot, cabbage, and lettuce ranged from 91 to 118% following fortification at 0.05-1.0 ppm. The detection limit of C2 in crops was 0.01 ppm (S/N > 3). The fortified samples of onion, sweet potato, kidney bean, and carrot were confirmed by LC/MS (ESI), and the peak of C2 was detected. PMID- 11501922 TI - Determination of aluminium and chromium in slurried baby food samples by electrothermal atomic absorption spectrometry. AB - Electrothermal atomic absorption spectrometry was used to determine aluminium and chromium in different types of baby foods. Samples without previous treatment were introduced into the atomizer as suspensions, which were prepared in a medium containing 0.1% (w/v) Triton X-100 (for chromium) or 10% (v/v) ethanol (for aluminium), 5% (v/v) concentrated hydrogen peroxide, and 0.5% (v/v) concentrated nitric acid. The slurries were homogenized with a potter and introduced directly into the furnace. The presence of nitric acid allowed a substantial amount of analyte to be extracted to the liquid phase. No matrix effect was observed and aqueous standard solutions were used for calibration. The detection limits were 4 and 50 pg for chromium and aluminium, respectively; characteristic masses were 3.8 pg for chromium and 18 pg for aluminium. Reliability of the procedures was checked by statistically comparing the results with those obtained with a previous microwave oven mineralization stage and by analyis of several certified reference materials. Values for the aluminium content ranged between 0.4 and 3.0 microg/g, and for chromium between 0.003 and 0.06 microg/g. PMID- 11501921 TI - Identical flow injection spectrophotometric manifold for determination of protein, phosphorus, calcium, chloride, copper, manganese, iron, and zinc in feeds or premixes. AB - A simple procedure using an identical manifold was developed for determination of nitrogen (protein) phosphorus, calcium, chloride, copper, manganese, iron, and zinc in feeds and feedstuffs. By changing appropriate reagents and detection wavelength, these 8 elements were determined successively with a simple identical double-line flow injection (FI) manifold. Fl spectrophotometric determinations were made by the blue indophenol reaction for ammonium, the molybdenum blue method for phosphate, the cresolphthalein complexone procedure for calcium, and the mercuric thiocyanate procedure for chloride. The chromogenic reagents for copper, iron, manganese, and zinc determination were bis(cyclohexanone)oxalydihydrazone (Cuprizone), 1,10-phenanthroline, formaldoxime, and xylenol orange, respectively. Sample digestion catalyst, Fl manifold, and some chemical parameters were optimized. The proposed procedure had a sampling rate of 90/h for each analyte. The determination ranges (mg/L) were 10 60 for N, 1-15 for P and Ca, 540 for Cl, and 0.5-15 for Cu, Fe, Mn, and Zn, respectively. Results of the analyses of animal feed and feedstuff samples by this procedure did not differ significantly from those obtained by proven manual methods. PMID- 11501923 TI - Determination of organochlorine pesticides and chlorobenzenes in strawberries by using accelerated solvent extraction combined with sorptive enrichment and gas chromatography/mass spectrometry. AB - An analytical scheme for the determination of several organochlorine pesticides like hexachlorocyclohexanes (HCHs) and DDX compounds (p,p'-DDE, p,p'-DDD, and p,p'-DDT) as well as chlorobenzenes in strawberries has been developed. The procedure is based on aqueous accelerated solvent extraction (ASE) followed by solid-phase microextraction (SPME) or stir bar sorptive extraction (SBSE) and subsequent thermodesorption-gas chromatography/mass spectrometry analysis. A 65 microm polydimethylsiloxane/ divinylbenzene fiber was chosen for the SPME experiments. Significant SPME and ASE parameters were optimized using spiked water and strawberry samples. For the ASE of the organochlorine compounds, a water-acetone mixture (90 + 10, v/v) as the extraction solvent, an extraction temperature of 120 degrees C, and 2 cycles of 10 min extraction proved optimal. The developed method was evaluated with respect to precision and limits of detection (LOD). The relative standard deviations of replicate ASE-SPME determinations (n = 5) were in the range of 4-24%. LOD values between 1 and 10 microg/kg were achieved with the exception of DDT and DDE (40 microg/kg). Using SBSE, the LOD of these compounds could be improved (2 and 5 microg/kg). The main advantages of this method are the avoidance of cleanup and concentration procedures as well as the significant reduction of the required volume of organic solvents. The described method was applied to the determination of the pollutants in strawberry samples collected from different allotment gardens in a potentially polluted area, the Bitterfeld-Wolfen region (Germany). PMID- 11501924 TI - Certified reference material for analytical quality assurance of minor and trace elements in food and related matrixes based on a typical Japanese diet: interlaboratory study. AB - A Certified Reference Material (CRM) was prepared at the National Institute for Environmental Studies (NIES), Japan, in collaboration with the National Institute of Radiological Sciences (NIRS), Japan, for the analytical quality assurance of minor and trace elements in food and related matrixes. The starting material for the CRM was all food served in 29 households in Japan over two 3-day periods in 1997-1998, and thus the CRM represented a typical Japanese diet. All foods (meals, snacks, and beverages) were homogenized, freeze-dried, pulverized, blended, dispensed into 1,100 bottles, and sterilized. The within- and between bottle homogeneity of the prepared CRM was satisfactory for most of the elements. The concentrations of 14 elements (Na, Mg, K, Ca, Mn, Cu, Zn, As, Se, Sr, Cd, Sn, Ba, and U) were certified based on a collaborative analysis involving NIES, NIRS, and 20 other laboratories. Reference values were given for the concentrations of 12 additional elements (P, Cl, Fe, Co, Ni, Br, Rb, Mo, I, Cs, Pb, and Th). The elements certified and those given reference values include minerals, essential trace elements, contaminant elements, and long-lived radionuclides. Thus, this CRM is of practical value in the quality assurance of element analysis of foods and diets in nutritional, environmental, and radiological research. PMID- 11501925 TI - Multiresidue determination of pesticides in fruit and vegetables by gas chromatography/tandem mass spectrometry. AB - Pesticide residues in fruit and vegetables were determined by gas chromatography/tandem mass spectrometry (GC/MS/MS). Electron impact (EI)/MS/MS and chemical ionization (CI)/MS/MS were developed for 80 compounds, including organochlorine, organophosphorus, organonitrogen, and pyrethroids, providing unambiguous spectral confirmation for these complex matrixes. Residues were extracted from samples with acetone followed by a mixture of dichloromethane petroleum ether. Two injections per sample were required for analysis of the entire pesticide list by EI/MS/MS and CI/MS/MS. Initial steps involving cleanup and concentration of extracts were eliminated. The excellent selectivity and good linearity allowed quantification and identification of low levels of pesticides in the most difficult matrixes. The method has been used for routine analysis of many vegetables. PMID- 11501926 TI - The role of planar chromatography in medicinal plant research. AB - This paper summarizes the role of planar chromatography (PC) in medicinal and aromatic plant (MAP) research and development, and demonstrates the importance of the technique, after extraction, in the analysis of MAP (identification and quantitative determination of the separated compound/s), in the purification and isolation process, and in different types of screening procedure. Special attention is paid to analytical, micropreparative and preparative forced-flow techniques, for example overpressured-layer chromatography (OPLC) and rotation planar chromatography (RPC). The special features of analytical, micropreparative, and preparative layer chromatography (PLC), OPLC, and RPC are compared in tables. Purification and isolation procedures using forced-flow techniques are shown in flowcharts. Some applications, relating to different classes of substance, are presented to demonstrate the versatility of various planar chromatographic techniques. PMID- 11501927 TI - Fluorophotometric thin-layer chromatography of ginkgo terpenes by postchromatographic thermochemical derivatization and quality survey of commercial ginkgo products. AB - Terpenoid lactones in Ginkgo biloba leaves are the main active constituents and the content of these terpenes is therefore the key factor for evaluating the quality of the leaves, the extract, and its preparations distributed on the market. The precleanup sample solutions were applied onto the silica gel plate modified with sodium acetate solution and developed with a solvent system of toluene-ethyl acetate-acetone-methanol, and a fluorescence chromatogram was generated by means of postchromatographic thermal chemical reaction. Fluorescence scanning was conducted quantitatively. The methodology validation confirmed that it is a practical alternative for routine quality control for ginkgo terpenes and the results are comparable with those obtained by high-performance liquid chromatography (HPLC). On the basis of the method established, a quality survey of the various commercial ginkgo products from different sources was undertaken. The obtained data demonstrated that the fluctuation of the content of individual terpene and/or the total terpenes among replicate samples is so significant as to cast doubts on the consistency of their pharmacological and clinical efficacy. PMID- 11501928 TI - Thin-layer chromatography--a useful technique for the separation of enantiomers. AB - Separation of enantiomers has become a well-established technique in many fields of science over the last decade. Unfortunately, even though there are a large number of chiral stationary phases able to perform enantiomeric separation, there is still a great deal of trial and error in developing a method for the separation of enantiomers. Thin-layer chromatography is a very versatile technique, which has brought much advancement in various fields of science. The simplicity of the technique makes it amenable for separation of enantiomers. This paper will present a review of the literature concerning separation of enantiomers. Because of the process of trial and error present in developing a chiral separation method, this paper also presents the mechanism underlying each form of separation. Thus, the methods are presented according to the main mechanism governing the particular separation. PMID- 11501929 TI - Plasma analysis of celiprolol by HPTLC: a useful technique for pharmacokinetic studies. AB - A rapid and sensitive high performance, thin-layer chromatographic (HPTLC) method has been developed for the measurement of celiprolol in human plasma and its use in pharmacokinetic studies has been evaluated. Detection and quantitation were performed without using an internal standard. A simple extraction procedure was followed for extracting celiprolol from plasma and a known amount of the extract was spotted on precoated silica gel 60 F254 plates using a Camag Linomat IV autosampler. Celiprolol was quantitated using a Camag TLC Scanner 3. The average recovery of authentic analytes (20 to 200 ng/mL) added to plasma was 72.06 +/- 2.8% and the lowest amount of celiprolol that could be detected was 10 ng/mL. The method provides a direct estimate of the amount of celiprolol present in plasma. Pharmacokinetic parameters of 2 marketed preparations have also been determined after oral administration to 12 healthy human volunteers. PMID- 11501930 TI - TLC determination of mebendazol and pentoxifylline as residues on pharmaceutical equipment surfaces. AB - This paper presents the applicability of thin-layer chromatographic methods with a subsequent densitometric or video densitometric quantitation for determination of residues in controlling pharmaceutical equipment cleanliness. Analytical methods were developed for monitoring residues of pentoxifylline at 10 mg/M2 and mebendazol at 1 mg/M2 on stainless steel surfaces. Simulated samples were prepared by addition of a calculated amount of pharmaceutical (as a solution) on a 35 x 35 cm stainless steel surface. After evaporation of solvent, the residues were wiped with wetted cotton. The cotton was extracted with dichloromethan methanol (1 + 1). Filtered extract was concentrated by vacuum evaporation and an aliquot applied to the plate, where standards were also applied. In the narrow concentration range near the acceptable residue limits, linear calibration curve could be obtained for both substances. The mean recovery (n = 4) obtained by densitometric quantitation was 93.4% for pentoxifylline and 85.6% for mebendazol, with coefficients of variation of 3.5 and 8.3%, respectively. Results of video densitometric quantitation did not differ significantly. However, data acquisition and evaluation is faster compared with densitometry and allows better archiving possibilities as required by the regulatory authorities. Both quantitation modes can be applied to routine control of pharmaceutical equipment cleanliness. PMID- 11501931 TI - Validation and quality assurance of planar chromatographic procedures in pharmaceutical analysis. AB - Within the process of the International Conference on Harmonization (ICH), 2 guidelines were released containing a standardized terminology, a verified model of requirements for the validation of analytical procedures, and some guidance in the practical aspects of conducting validation studies in pharmaceutical analysis. For planar chromatographic procedures, which may be used at different levels either in qualitative identity testing, assays, semiquantitative limit tests, or quantitative determination of impurities, this paper tries to transfer these formal requirements into practical approaches for validation. Basic acceptance criteria for evaluation of validation experiments based on practical experience are proposed. In addition, selected parameters for robustness testing of given procedures and quality assurance of quantitative planar chromatographic testing by control charts is described. PMID- 11501932 TI - Separation and quantitation of alkylphosphocholines and analogues of different liposome formulations by HPTLC. AB - High-performance thin-layer chromatographic (HPTLC) analysis of non UV-active phospholipids in biological matrixes is a common method for separation, detection, and quantitation. Liposomes containing new alkylphosphocholines and analogues with enhanced cytostatic activity had been prepared. The liposomal formulations were designed to enable the intravenous application of the alkylphosphocholines and analogues and to reduce dose-limiting toxicities observed after oral administration. For quality control the liposomes were analyzed by HPTLC for content of 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG), cholesterol, alkylphosphocholines, and analogues and their related compounds (main degradation products). Due to the differences in lipophily of the compounds, different mobile phases were necessary to achieve separation. Automated Multiple Development was used to reduce the number of plates and to improve the selectivity and the capacity of the chromatographic system to separate the described alkylphosphocholines and analogues from DPPG and 1,2 dipalmitoyl-sn-glycero-3-phosphocholine in one chromatographic system. PMID- 11501933 TI - Quantitative determination of phospholipids in a pharmaceutical drug by scanning and video densitometry. AB - This paper describes a convenient and practice method for quantitation of surfactant phospholipids (1,2-dipalmitoyl-3-sn-phosphatidyl choline [DPPC] and 1 palmitayl-2-oleyl-3-sn-phosphatidyl glycerol [POPG]) in a recombinant surfactant lyophile (Venticute) by high-performance, thin-layer chromatography (HPTLC) with video densitometry. DPPC and POPG were extracted from Venticute-lyophile using methanol. Separation from the other active ingredients and excipients was accomplished by HPTLC on silica gel F254 plates with a mixture of chloroform, methanol, glacial acetic acid, and water as development solvent. Postchromatographic derivatization by dipping in copper sulphate/phosphoric acid reagent and subsequent heating shows grey-brown bands on a light blue background. These were detected with the video densitometer in the VIS range, and with scanning densitometry at 365 nm. Linear calibration in a working range of 0.7-1.3 microg DPPC and 0.35-0.65 microg POPG was demonstrated by integrating the area under the peaks. Good results were obtained with recovery experiments. When compared to classical slit scanning densitometry, video densitometry represents a fast alternative to quantitate thin-layer chromatograms in surfactant phospholipid analysis. PMID- 11501934 TI - Instrumental planar chromatographic determination of benzodiazepines: comparison with liquid chromatography and gas chromatography. AB - Diazepam, chlordiazepoxide, and midazolam are determined by high-perfromance, thin-layer chromatography (HPTLC). Results are compared with those obtained by liquid chromatography (LC) and gas chromatography (GC). Analytical conditions for all the methods are also described. Validation parameters for linearity, precision, detection, and quantitative limits are informed. Correlation study of HPTLC with LC and GC gave a correlation factor between 0.98 and 0.99. The obtained results showed that HPTLC, LC, and GC techniques are comparable for determination of such benzodiazepins in the requested working range to be analyzed in raw materials. PMID- 11501935 TI - Application of high-performance, thin-layer chromatography to quality control of antimetabolite analogue infusion bags. AB - A global postproduction quality program was developed to secure chemotherapy infusion at the Gustave Roussy Institute. Despite rigorous procedures and computerized prescriptions, an analytical check was necessary to improve the quality of ready-to-use solutions of cytotoxic drugs in our Centralized Antineoplastics Reconstitution Unit. High-performance, thin-layer chromatography was selected as the analytical tool to assay 12 anticancer drugs. One of the analytical methods can separate 4 antimetabolite substances, i.e., fludarabine (FDB), cytarabine (CTB), gemcitabine (GTB), and fluorouracil (5 FU). For all infusion bags manufactured, up to 26 samples could be assayed per series using a double standard calibration (GTB and 5 FU). PMID- 11501936 TI - Horwitz receives Boyle Medal from Royal Society of Chemistry. PMID- 11501937 TI - Pohland receives Professional Service Award from Alpha Chi Sigma's Washington, DC, Chapter. PMID- 11501938 TI - Recent advances in thin-layer chromatography of pesticides. AB - Advances in the applications of thin-layer chromatography (TLC) and high performance thin-layer chromatography (HPTLC) for the separation, detection, and qualitative and quantitative determination of pesticides, other agrochemicals, and related compounds are reviewed for the period 1998-2000. Analyses are covered for a variety of samples, such as food, biological, and environmental, and for residues of pesticides of various types, including insecticides, herbicides, and fungicides, belonging to different chemical classes. References on formulation analysis, hydrophobicity studies, and the use of TLC and thin-layer radiochromatography (TLRC) for studies of pesticide metabolism, degradation, uptake, and related studies are also included. PMID- 11501939 TI - Corneal dystrophies in Japan. AB - Recent advances in molecular genetics have increased our understanding of the role of genes. Four autosomal dominant corneal dystrophies (CDs); granular CD (GCD), Avellino CD (ACD), lattice CD (LCD), and Reis-Bucklers CD (RBCD) were mapped to the long arm of chromosome 5 (5q31). These four diseases were shown, in a Caucasian series, to result from different missense mutations in the TGFBI (BIGH3, keratoepithelin) gene. The same mutations were also detected in Japanese patients, from a different ethnic background. Gelatinous drop-like corneal dystrophy (GDLD), on the other hand, which was found in Japanese patients in 1914, is a rare autosomal recessive disorder characterized by corneal amyloidosis. Parents of the patients had a markedly higher frequency of consanguineous marriages than the general population. The gene responsible for GDLD, the membrane component, chromosome 1, surface marker 1 (M1S1) gene was mapped to the short arm of chromosome 1(1p). Four deleterious mutations in this gene were detected in Japanese patients. We review here additional studies on mutations of the TGFBI and M1S1 genes found in Japanese patients. In the TGFBI gene, nine different mutations were detected in Japanese patients with GCD, ACD, LCD, or RBCD. The codons R124 and R555 of the TGFBI gene were hotspots in Japanese patients, of whom many were ACD patients with the R124H mutation. New mutations responsible for LCD were detected in the TGFBI gene of patients with LCD, in addition to the P501T mutation in LCD type IIIA found earlier. These studies showed a clear genotype/phenotype correlation associated with the TGFBI gene. In the M1S1 gene, the Q118X mutation was the most common alteration, and a founder mutation in Japanese GDLD patients, as previously reported. Ninety-two percent of the mutated alleles were the Q118X. PMID- 11501940 TI - Association of a G994 --> T (Val279 --> Phe) polymorphism of the plasma platelet activating factor acetylhydrolase gene with myocardial damage in Japanese patients with nonfamilial hypertrophic cardiomyopathy. AB - Plasma platelet-activating factor acetylhydrolase (PAF-AH) acts as a key defense against oxidative stress by hydrolyzing PAF and oxidized phospholipids. Deficiency of the activity of this enzyme may thus potentially result in predisposition to myocardial damage. The possible role of the G994 (V allele) --> T (F allele) polymorphism of the PAF-AH gene in modulating cardiac function was investigated in 142 Japanese subjects with nonfamilial hypertrophic cardiomyopathy (HCM). Logistic regression analysis adjusted for age, sex, height, and body weight revealed that the frequency of the F allele was significantly higher in HCM patients than in 284 healthy controls. Echocardiographic examination revealed that left ventricular (LV) end-diastolic and end-systolic dimensions were significantly greater in HCM patients with the FF genotype than in those with the VV genotype. Cardiac catheterization revealed that LV end diastolic pressure was significantly higher, whereas the LV ejection fraction was significantly smaller, for HCM patients with the F allele than for those with the VV genotype. Interstitial fibrosis was significantly more severe in HCM subjects with the FF genotype than in those with the VV genotype. These results suggest that the G994 --> T (Val279 --> Phe) polymorphism in the plasma PAF-AH gene may exacerbate cardiac damage in Japanese individuals with nonfamilial HCM, although this polymorphism is unlikely to be a causative factor for this condition. PMID- 11501941 TI - Association analysis of polymorphisms in the prepronociceptin gene and schizophrenia. AB - The human prepronociceptin (ppNoc) gene is located on chromosome 8p21, and several linkage studies have suggested that the susceptibility locus for schizophrenia is present in this chromosomal region. We investigated genetic variations in the promoter and coding regions of the ppNoc gene to determine if there may be an association between naturally occurring mutations and the manifestation of schizophrenia. Association analyses for the variations found were conducted between 87 unrelated schizophrenic patients and 100 healthy controls. Polymorphisms were found at both the -503 locus (G/A) in the promoter region and the 353 locus (Ala118Gly) in exon 3. Additionally, rare variants were identified at the -261 locus (A/G) in the promoter region, the 515 locus (Gln172Arg) in exon 3, and the 561 locus (G/A) in the untranslated region of exon 3. A significant difference was observed in allele frequency at the 353 locus between controls and patients with continuous-course schizophrenia (P = 0.0237), and between patients with and without prominent negative symptoms (P = 0.0201). However, neither difference was significant after Bonferroni correction. In addition, there were no significant differences in genotype and allele frequencies at either the -503 or 353 locus between the control group and patients with schizophrenia. These results suggest that ppNoc gene polymorphisms have no association with schizophrenia. PMID- 11501942 TI - Analysis of 168 short tandem repeat loci in the Japanese population, using a screening set for human genetic mapping. AB - We devised a multiplex polymerase chain reaction (PCR) amplification and loading system for the convenient typing of 168 short tandem repeat (STR) polymorphic markers in a commercially available screening primer set for human linkage analysis. We genotyped all these 168 STR loci with 32 healthy unrelated Japanese, calculated allele frequencies at each STR locus, and performed three kinds of tests for Hardy-Weinberg equilibrium (HWE). Significant deviations from HWE in all three tests were observed at only three loci, and the average heterozygosity in the Japanese (0.733) was slightly lower than that in Caucasians (0.773). We also examined 32 Caucasians at some selected loci, to be compared with Japanese. Some markers showed greatly different heterozygosities or allelic distributions in Japanese and Caucasian populations. In two groups of STRs, those with and without irregular alleles (or interalleles), the former had a higher proportion of bimodal allelic distribution and possessed more alleles per locus than the latter. However, no significant differences in the observed and expected heterozygosities, or in the powers of discrimination, were found between the two groups. The present basic study of allele frequency databases of these STRs will contribute to further applications in forensic science and human genetics. PMID- 11501943 TI - Identification of sequence polymorphisms of the COMP (cartilage oligomeric matrix protein) gene and association study in osteoarthrosis of the knee and hip joints. AB - Osteoarthrosis (OA) is a common cause of musculoskeletal disability characterized by late-onset degeneration of articular cartilage. Although several candidate genes have been reported, susceptibility genes for OA remain to be determined. Hereditary osteochondral dysplasias produce severe, early-onset OA and hence are models for common idiopathic OA. Among them are pseudoachondroplasia and multiple epiphyseal dysplasia, both of which are caused by mutations in the cartilage oligomeric matrix protein (COMP) gene. Therefore, COMP may be a susceptibility gene for OA. We screened for polymorphisms by direct sequencing of all exons of the COMP gene with their flanking intron sequences and the promoter region. We identified 16 polymorphisms, of which 12 were novel. Using six polymorphisms spanning the entire COMP gene, we examined the association of COMP in Japanese patients with OA of the knee and hip joints. Genotype and allele frequencies of the polymorphisms were not significantly different between OA and control groups, and there was no significant difference in haplotypes. These results do not support an association between COMP and OA in the Japanese population. PMID- 11501944 TI - Genomic organization and promoter analysis of human KCNN3 gene. AB - KCNN3 is a member of the gene family, KCNN1-4, encoding the small and intermediate conductance calcium-activated potassium channels. Long CAG-repeat alleles of this gene have been found to be over-represented in patients with schizophrenia in a number of population-based association studies, and this gene maps to human chromosome 1q21, a region recently implicated in schizophrenia by linkage. To set the stage for a further functional evaluation of KCNN3, we defined the nature of the genomic locus in the size, structure, and sequence of its introns and exons and the function of potential upstream regulatory regions. We isolated P1-derived artificial chromosome (PAC) clones from a genomic library and identified an overlapping available bacterial artificial chromosome (BAC) clone. Cosmids subcloned from the PAC and BAC clones were then sequenced and merged with the sequence in the public database. The KCNN3 gene spans over 163.1 kb and is composed of eight exons and seven introns. All of the exon-intron junctions conform closely to consensus splice sites. The proximal 2.5 kb of the 5'-flanking sequence was obtained and analyzed for potential transcription factor binding sites. In the proximal 2.5 kb upstream region, potential sites for the Ikaros factor (IK2), homeodomain factor Nkx-2.5/Csx (NKX25), nuclear factor of activated T-cells (NFAT), upstream stimulating factor (USF), c-AMP responsive element binding protein (CREB), POU factor Brn2 (BRN-2), myeloid zinc finger protein (MZF1), vitellogenin binding protein (VBP), HNF3 forkhead homologue 2 (HFH2), and transcription initiation were identified, as well as several potential AP-1 and AP-4 sites. Finally, a 2261-bp fragment of this upstream region was cloned into a promoterless pGL3-luciferase vector, where it produced orientation-dependent expression of the reporter gene in transiently transfected PC12 cells, cells which natively express functional KCNN3 channels, suggesting that this cloned fragment includes competent promoter elements of this gene. PMID- 11501945 TI - A high-throughput SNP typing system for genome-wide association studies. AB - One of the most difficult issues to be solved in genome-wide association studies is to reduce the amount of genomic DNA required for genotyping. Currently available technologies require too large a quantity of genomic DNA to genotype with hundreds or thousands of single-nucleotide polymorphisms (SNPs). To overcome this problem, we combined the Invader assay with multiplex polymerase chain reaction (PCR), carried out in the presence of antibody to Taq polymerase, as well as using a novel 384-well card system that can reduce the required reaction volume. We amplified 100 genomic DNA fragments, each containing one SNP, in a single tube, and analyzed each SNP with the Invader assay. This procedure correctly genotyped 98 of the 100 SNP loci examined in PCR-amplified samples from ten individuals: the genotypes were confirmed by direct sequencing. The reproducibility and universality of the method were confirmed with two additional sets of 100 SNPs. Because we used 40 ng of genomic DNA as a template for multiplex PCR, the amount needed to assay one SNP was only 0.4 ng; therefore, theoretically, more than 200,000 SNPs could be genotyped at once when 100 microg of genomic DNA is available. Our results indicate the feasibility of undertaking genome-wide association studies using blood samples of only 5-10 ml. PMID- 11501946 TI - The power of genome-wide association studies of complex disease genes: statistical limitations of indirect approaches using SNP markers. AB - Genome-wide association studies using a dense map of single nucleotide polymorphism (SNP) markers seem to enable us to detect a number of complex disease genes. In such indirect association studies, whether susceptibility genes can be detected is dependent not only on the degree of linkage disequilibrium between the disease variant and the SNP marker but also on the difference in their allele frequencies. These factors, as well as penetrance of the disease variant, influence the statistical power of such approaches. However, the power of indirect association studies is not well understood. We calculated the number of individuals necessary for the detection of the disease variant in both direct and indirect association studies with a case-control design. The result shows that a remarkable reduction in the statistical power of indirect studies, compared with that of direct ones, is unavoidable in the genome-wide screening of complex disease genes. If there is a large difference in allele frequency between the disease variant and the marker, the disease variant cannot be detected. Because the frequency of the disease variant is unknown, SNP markers with various allele frequencies, or a large number of SNP markers, must be used in indirect association studies. However, if the number of SNP markers is increased, the obtained P value may not reach the significance level due to the Bonferroni adjustment. Thus, to test a possible association between functional variants and a complex disease directly, we should identify such SNPs in as many genes as possible for use in genome-wide association studies. PMID- 11501948 TI - Single nucleotide polymorphisms of the fukutin gene. AB - Mutations in the LMNA gene, which encodes nuclear lamins A and C, underlie both Emery-Dreifuss muscular dystrophy (EMD2) and Dunnigan-type familial partial lipodystrophy (FPLD). This indicates that one gene can cause different phenotypes characterized by tissue degeneration. The gene for one form of Berardinelli-Seip type congenital total lipodystrophy (BSCL) has been mapped to chromosome 9q34. Based on the observation that one gene caused both FPLD and EMD2, we considered that a known gene for muscular dystrophy at or near the BSCL locus on chromosome 9q would be an appropriate candidate for BSCL. The gene encoding fukutin, which is mutated in Fukuyama congenital muscular dystrophy has been mapped to 9q31. We thus developed amplification primers for the coding regions of the fukutin gene. We found no putative disease mutations, but through screening of diseased and normal subjects, we identified three novel single nucleotide polymorphisms (SNPs). We conclude that mutations in fukutin are not present in subjects with BSCL. However, the identification of SNPs provides tools to investigate this protein for association with other phenotypes. PMID- 11501947 TI - Cloning and sequencing of a novel human gene that encodes a putative target protein of Nesh-SH3. AB - By using a conventional two-hybrid technique with an Src homology 3 (SH3) domain of Nesh as the bait protein, a novel full-length cDNA was isolated and sequenced from a human placenta cDNA library. This cDNA consists of 3023 bp and has a predicted open reading frame that encodes 486 amino acids. It possesses an SH3 binding motif, a nuclear targeting sequence, and no catalytic domain. Overall, it has no similarity to known molecules involved in a signaling cascade. Polymerase Chair reaction-based mapping with both a monochromosomal hybrid panel and radiation hybrid cell panels localized the gene on human chromosome 3q12 near the marker D3S1271. PMID- 11501949 TI - Single nucleotide polymorphisms of the very low density lipoprotein receptor (VLDLR) gene. AB - The very low density lipoprotein receptor (VLDLR) has a potentially important role in lipoprotein metabolism and Alzheimer's disease. We developed amplification primers for most of the coding region and 3'-untranslated region of VLDLR and used sequencing of genomic DNA to examine these regions of VLDLR in subjects with familial combined hyperlipidemia and in normal controls. We identified ten novel single nucleotide polymorphisms (SNPs) for VLDLR. We also found one rare coding sequence variant, S>R153, in a subject with familial combined hyperlipidemia, which was absent from 2360 normal alleles. The identification of intron-exon boundaries, amplification primers, and SNPs provides tools to investigate VLDLR for genetic association and linkage studies. PMID- 11501950 TI - Linkage disequilibrium and haplotype analysis among ten single-nucleotide polymorphisms of interleukin 11 identified by sequencing of the gene. AB - Interleukin (IL11) is a member of the interleukin 6 (IL6)-related cytokine subfamily, which stimulates T cell-dependent development of immunoglobulin producing B cells. IL11 is also an important paracrine regulator of bone metabolism that induces formation of osteoclasts. In the work reported here, we sequenced the entire IL11 structural gene of 48 alleles in a Japanese test population. These experiments identified ten single-nucleotide polymorphisms (SNPs) and determined their allelic frequencies. One polymorphism was identified upstream of exon 1, one in exon 3, four in intron 4 and four in the 3' untranslated region (3'UTR) of exon 5. Based on the genotype data, we constructed six haplotypes in the tested population. Two-way comparisons of SNPs revealed two combinations in complete linkage disequilibrium, one with SNPs at nucleotide positions 2753, 3644, 5154, and 5568, and another with SNPs at positions 3686, 5141, and 5734. These results will be useful in disease-association studies where a contribution of the human IL11 gene has been suspected, especially in disorders affecting immune response and bone metabolism. PMID- 11501951 TI - Quality control in cardiovascular research. PMID- 11501952 TI - Quality assurance in cardiovascular medical publications. PMID- 11501953 TI - Quality control in cardiovascular research: a difficult task. PMID- 11501954 TI - The search for the causes of myocardial infarction. Introduction. PMID- 11501955 TI - Laboratory markers of hypercoagulability. AB - Investigations carried out over the last 40 years have demonstrated that coronary artery thrombosis is the critical event underlying myocardial infarction and unstable angina. The existence of a prolonged hypercoagulable state preceding the thrombotic event has been postulated for some time and significant associations have been established between the plasma concentrations of a number of hemostatic variables and the frequency of myocardial infarction. High plasma fibrinogen, factor VII/VIIa, tissue-type plasminogen activator and plasminogen activator inhibitor levels have been associated with at least as great a risk of developing myocardial (re)infarction or sudden death as high cholesterol levels, especially in the young. In the last year more sensitive assays have been developed, and they should allow a precise biochemical definition of hypercoagulable states. The significance of these new assays and their role in defining a hypercoagulable state in different conditions are analyzed. PMID- 11501956 TI - Prothrombotic genetic markers. AB - The last decade has been characterized by an explosion of research studies on genetic epidemiology. In particular, as far as ischemic heart disease is concerned, a lot of research was focused on prothrombotic genetic risk factors. Unfortunately, the success of this approach in the field of venous thrombosis has not been replicated in the field of myocardial infarction. In the present editorial, a comment on the studies already available is provided and the possible limitations of the present approach are analyzed. PMID- 11501957 TI - Hemostatic markers and prognosis in ischemic heart disease. AB - Failure of traditional risk factors in identifying patients who develop a cardiac event, has led investigators to focus on other factors involved in precipitating cardiac events. As acute or subacute thrombosis is the major complication of atherosclerotic plaque rupture, attention has been dedicated to prothrombotic markers as possible risk factors. Recently, the role of new laboratory markers in predicting the risk of cardiac events has been evaluated in large epidemiological studies. The results of these studies as well as the value and applicability of new prothrombotic markers in the clinical practice are discussed. PMID- 11501958 TI - The A-Med right heart support for off-pump coronary artery bypass grafting. AB - BACKGROUND: Off-pump coronary artery bypass grafting (OPCAB) presents several advantages but, mainly due to the impaired diastolic filling of the right ventricle, the displacement of the heart can cause hemodynamic instability. The aim of this study was to investigate the possible role of the A-Med right heart support during OPCAB. METHODS: We report our early experience with the A-Med system (A-Med, West Sacramento, CA, USA) during OPCAB. The system consists of a coaxial cannula, a microcentrifugal pump and a control console. The coaxial cannula is passed through the right atrium with the tip of the cannula positioned in the main pulmonary artery. Thus the blood is actively removed from the right atrium and returned to the pulmonary artery. RESULTS: We successfully used this right heart support in 2 patients undergoing elective OPCAB. In both cases the system was used during the exposure of the proximal portion of the obtuse marginal branch. A mean pump flow of 3.2 l/min guaranteed normal cardiac output and hemodynamic stability during the exposure of the posterior target area. No complication occurred and the patients were discharged shortly after surgery. CONCLUSIONS: In our early experience the A-Med right heart support was safe and effective and allowed achievement of hemodynamic stability during exposure of the posterior areas of the left ventricle. PMID- 11501959 TI - Risk predictors of paroxysmal atrial fibrillation following aortic valve replacement. AB - BACKGROUND: Atrial fibrillation (AF) is the most frequently encountered arrhythmic complication associated with cardiac surgery. The aim of this paper was to identify the clinical predictors of AF occurrence following aortic valve replacement. METHODS: Three hundred and two patients were included in this study and divided into two groups according to the absence (SR group, 243 patients, mean age 55.6 +/- 15 years) or the evidence (AF group, 59 patients, mean age 63.8 +/- 11 years) of post-aortic valve replacement AF. Sixty-five perioperative variables (37 preoperative, 8 intraoperative and 20 postoperative) were considered. RESULTS: Post-aortic valve replacement paroxysmal AF occurred in 59 out of 302 patients (19%). At univariate analysis, post-aortic valve replacement AF was associated with advanced age, left atrial enlargement, preoperative episodes of paroxysmal AF, the use of a warm blood cardioplegic solution and normothermia, administration of inotropic agents, prolonged assisted ventilation but also with postoperative acidosis, electrolyte imbalance and atrioventricular and intraventricular conduction disorders. Stepwise forward multivariate logistic regression analysis identified age (p = 0.002, odds ratio--OR 1.04), left atrial enlargement (p = 0.004, OR 2.6), a prior history of paroxysmal AF (p = 0.0003, OR 10.9), and postoperative electrolyte imbalance (p = 0.01, OR 2.3) as independent correlates of AF, whereas the use of hypothermia appeared to be a protective factor (p = 0.0004, OR 0.26). CONCLUSIONS: According to our findings, post-aortic valve replacement AF seems to be associated with well-defined anatomical and electrical substrates generated by advanced age, increased left atrial dimensions, and a possible electrical remodeling consequent to prior repetitive episodes of paroxysmal AF. On these grounds, external factors such as postoperative electrolyte imbalance might enhance atrial ectopic activity and trigger postoperative sustained tachyarrhythmias, while the use of hypothermia might allow for better protection of the atrial myocardium against intraoperative ischemia. PMID- 11501960 TI - Can short-term verapamil therapy reduce the recurrence of atrial fibrillation after successful low energy intracardiac cardioversion? AB - BACKGROUND: Calcium-lowering drugs seem to be able to reduce the recurrences of atrial fibrillation (AF) after cardioversion by preventing electrical remodeling of atrial cells. The aim of our study was to prospectively evaluate the efficacy of short-term verapamil therapy associated with propafenone or amiodarone in reducing recurrences of AF after low energy intracardiac cardioversion. METHODS: Eighty-two patients with chronic AF (mean duration 6.1 months, range 1-96 months) underwent low energy intracardiac cardioversion. Forty-one patients (Group A) were instructed to suspend antiarrhythmic therapy 48 hours before the procedure (only chronic amiodarone was allowed). The subsequent 41 patients (Group B), in addition to previous prescriptions, had to take verapamil (120 mg twice daily) for 3 days before low energy intracardiac cardioversion and for 7 days after cardioversion. A right atrium-coronary sinus or right atrium-left pulmonary artery electrode configuration was indifferently utilized. Propafenone (450-900 mg daily) or amiodarone (200 mg daily) was prescribed to all patients after cardioversion. RESULTS: Sinus rhythm was acutely restored in 80 patients (97.6%): the mean number of shocks delivered was 2.3 (range 1-5); the mean energy required was 10.5 J (range 7.2-19.8 J). No statistically significant differences were found between the right atrium-coronary sinus vs right atrium-left pulmonary artery electrode configuration regarding the energy required and the number of shocks delivered. Group A and Group B showed the same number of AF recurrences at the first month of follow-up. CONCLUSIONS: In our study, short-term verapamil treatment associated with propafenone or amiodarone seems to be useless for the prevention of recurrent AF after low energy intracardiac cardioversion. PMID- 11501961 TI - Characterization of post-discharge atrial fibrillation following open-heart surgery in uncomplicated patients referred to an early rehabilitation program. AB - BACKGROUND: Atrial fibrillation (AF) is frequently observed after open-heart surgery, following discharge from the cardiac surgery clinic. Compared to those usually reported in the early postoperative period, this arrhythmia is delayed in onset and is often a cause of re-hospitalization. Post-discharge AF has never been characterized in the literature. METHODS: We retrospectively analyzed post discharge AF occurring within 30 days of coronary artery bypass graft or of valvular procedures in 376 patients referred to an early postoperative rehabilitation program. To investigate the probability of the persistence of post discharge AF, we prospectively examined 232 patients who had undergone valvular procedures. RESULTS: An arrhythmia was recognized in 61/376 patients (16%), resulted in worsening of the NYHA functional class in 27 (44%) and in life threatening hemodynamic effects requiring urgent cardioversion in 5 (8%). Events were predicted by the occurrence of postoperative AF (6-fold higher risk), left ventricular hypertrophy, an enlarged left atrium and valvular pathology (3-fold) and by the lack of beta-blocker protection (5-fold). AF persisted in 20/232 (9% of the study population, 18% of patients who had post-discharge AF) and had a relevant impact on the patient's clinical status. Predictors of events were older age, an enlarged left atrium and a lower left ventricular ejection fraction. CONCLUSIONS: Post-discharge AF following open-heart surgery is frequent in patients undergoing valvular procedures and often persists over time. The clinical impact of the arrhythmia is relevant, it might cause re-hospitalizations in many circumstances and, consequently, may have an impact on hospital resources. Events are much less frequent in patients taking beta-blockers than in those who do not, and they can be predicted by simple variables observed in the early stages after surgery. PMID- 11501962 TI - Time-domain analysis of exercise-induced ST-segment elevation in Q-wave myocardial infarction: a useful tool for the screening of myocardial viability. AB - BACKGROUND: Exercise-induced ST-segment elevation in Q-wave leads has been traditionally associated with passive stretching of the infarct wall, perinecrotic ischemia and, according to recent scintigraphic studies, with myocardial viability. At present, however, no definitive conclusions are available. We evaluated the potential role of a time-domain analysis of exercise induced ST-segment elevation for the identification of viable myocardium and residual ischemia in patients with previous Q-wave myocardial infarction. METHODS: Sixty patients with a previous Q-wave myocardial infarction underwent a bicycle exercise stress test, dobutamine stress echocardiography, coronary arteriography and left ventriculography. RESULTS: Patients with exercise-induced ST-segment elevation in Q-wave leads (n = 36) showed more severe impairment of resting left ventricular function, when evaluated in terms of wall motion score index at echocardiography (1.62 +/- 0.33 vs 1.41 +/- 0.22, p < 0.01) and in terms of wall motion score at ventriculography (5.9 +/- 1.6 vs 4.1 +/- 1.5, p < 0.03), compared to patients without ST-segment shift (n = 24). No differences between the two groups were seen in the severity and extension of coronary artery disease. The two groups of patients did not differ in the overall incidence of viability (50% in patients with vs 62% in those without ST-segment elevation, p = NS) and homozonal ischemia (39 vs 26%, p = NS), when evaluated with dobutamine echocardiography. However, a time-domain analysis of the ST-segment changes during exercise showed that the duration of exercise up to 0.1 mV ST-segment elevation was significantly lower in patients with viability (6.2 +/- 3.3 min) than in those without (10.2 +/- 2.2 min) (p < 0.001). Accordingly, ST-segment elevation occurred within 3 and 6 min of exercise in 7/18 and in 12/18 patients with viability respectively, but in only 0/18 (p < 0.01) and in 1/18 (p < 0.01) patients without viability. Thus, ST-segment elevation occurring within the first two stages of the exercise test was, respectively, 39 and 67% sensitive and 100 and 94% specific for viability. Early onset ST-segment elevation (within 3 and 6 min) was also more frequent in patients with high-dose dobutamine-induced homozonal ischemia than in those without (sensitivity for ischemia 50 and 67%; specificity 95 and 74%, respectively). CONCLUSIONS: After myocardial infarction, ST-segment elevation in Q-wave leads at the peak of exercise is associated with severe resting left ventricular dysfunction but fails to identify patients with a viable myocardium or residual ischemia. Instead, ST-segment elevation occurring in the early phases of exercise is a highly specific, although not very sensitive marker of dobutamine-assessed viability in the infarct area and may be indicative of residual ischemia. PMID- 11501963 TI - Diagnostic accuracy and prognostic implications of stress testing for coronary artery disease in the elderly. AB - BACKGROUND: The aim of this study was to define the diagnostic accuracy and the prognostic significance of stress electrocardiography (ECG) and of thallium-201 single-photon emission computed tomography (SPECT) in determining the incidence of coronary artery disease (CAD) in an elderly population. METHODS: A selected series of 132 patients (90 males, mean age 72.4 years; 42 females, mean age 68.2 years) hospitalized because of cardiac events associated with suspected CAD, underwent stress ECG and thallium-201 testing; as endpoints we considered the heart rate and the appearance of clinical symptoms or ST segment depression. Patients unable to develop an adequate exercise workload, were tested with dipyridamole. All patients also underwent coronary angiography following the stress test. One hundred and twenty-four patients had a mean follow-up of 2 years. Endpoints included subsequent coronary events or new serious disease. The sensitivity, specificity, diagnostic accuracy, and positive and negative predictive values of both ECG and of thallium-201 SPECT were calculated. RESULTS: ECG findings were positive in 102 patients and coronary angiography confirmed the presence of CAD in 92 of them; ECG findings were negative in 30 patients, but only 14 were free from obstructive coronary lesions. Thallium-201 SPECT findings were positive in 112 patients (coronary angiograms confirmed CAD in 101) and negative in 20 patients (in 13 of whom angiography confirmed the absence of disease). The sensitivity of the stress tests was quite good: 85.1% for ECG and 93.5% for thallium-201 SPECT; conversely, the specificity of ECG was superior to that of SPECT (58.3 vs 54.1%). The sensitivity of both techniques was superior in males than in females and seemed to correlate with the extent of CAD. CONCLUSIONS: The diagnostic accuracy of thallium-201 SPECT was 86.3% whereas that of ECG was 80.3%. Considering the overall cardiac events, the predictive value of SPECT was superior to that of ECG both in terms of the positive value (54 vs 51%, p = NS) and, more importantly, in terms of the negative value (84 vs 62%, p < 0.03). In fact, patients with normal thallium images were at low risk for future cardiac events. PMID- 11501964 TI - Role of diet and lifestyle in the prevention of cardiovascular disease: knowledge and attitude of physicians and patients in Southern Italy. AB - BACKGROUND: The present study aimed to investigate the knowledge and attitude of family physicians and their patients respectively in prescribing and adopting lifestyle and dietary changes in order to prevent cardiovascular disease which is the main cause of mortality in the industrialized Western countries. METHODS: All physicians working in two suburban areas of Naples and in the city of Avellino were invited to participate in the study and 63% accepted (n = 218). A sample of lay people living in the same areas was randomly selected and 59% accepted (n = 267). Two different questionnaires were used to assess the knowledge and attitudes of physicians and lay people about the role of nutrition and lifestyle in the prevention of cardiovascular risk. RESULTS: Physicians showed satisfactory knowledge and a good attitude to non-pharmacological prevention--particularly secondary prevention--of cardiovascular disease. The opinions of the lay people interviewed on the impact of lifestyle changes on cardiovascular disease prevention were similarly satisfactory although some inconsistencies were noted. CONCLUSIONS: In spite of the good propensity of both physicians and their patients to the implementation of lifestyle changes, it appears that dietary modifications prescribed by physicians are not accurately followed or even misunderstood by their patients. This suggests the need of implementing further educational programs. PMID- 11501965 TI - Constrictive pericarditis after cardiac surgery. AB - Constrictive pericarditis is an infrequent complication of cardiac surgery. We report the case of a young woman who developed dyspnea and ascites 3 years after surgical closure of an atrial septal defect, and the findings at chest X-ray, computed tomographic scan and Doppler echocardiography are described. Epidemiology of the disease, new pathophysiologic concepts, diagnostic features, and therapeutic targets are reviewed. PMID- 11501966 TI - Heart metastases from gastric sarcoma. A case report. AB - Cardiac involvement by metastatic gastric sarcoma is a rare event. We describe an unusual case of gastric sarcoma involving the heart in a 29-year-old man who presented with heart failure. The diagnosis was made at two-dimensional echocardiography. Heart metastases prolapsing into and obstructing the mitral valve orifice were located in the left atrium. In an attempt to excise the left atrial mass, the patient underwent heart surgery twice. The resections were non curative. The patient died of refractory heart failure 9 months after the onset of symptoms. This case report is an example of a secondary intracavitary tumor that causes obstruction of the mitral valve orifice. Two-dimensional echocardiography has been confirmed as the examination of choice for the early diagnosis of cardiac tumors. PMID- 11501967 TI - Chylothorax: a complication after internal thoracic artery harvesting. AB - Chylothorax is a rare but serious complication of cardiac surgery. A 64-year-old man with three-vessel disease underwent coronary artery bypass grafting. Ten days later he developed left pleural effusion. An intercostal drain was inserted and 1600 ml of pale pink, milky fluid were obtained. The results of biochemical analysis were consistent with chyle. The diagnosis of a left chylothorax was made. Conservative treatment consisting of total parenteral nutrition and pleural drainage was successfully employed. In the literature we found 17 cases in which the development of chylothorax after a coronary revascularization procedure is described. PMID- 11501968 TI - ATRA(ouble) in the treatment of acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is a unique disease that responds to differentiation-inducing effects of all-trans-retinoic acid (ATRA). ATRA induces complete clinical remissions (CRs) in most patients and now constitutes a standard therapy in patients with APL. However, CRs induced by ATRA are usually brief, and resistance to the therapy rapidly develops, leading to relapses in almost every patient; thus limiting the use of ATRA as a single agent. On the basis of clinical and in vitro studies, the following mechanisms have been proposed to explain ATRA resistance: 1) induction of accelerated metabolism of ATRA, 2) increased expression of cellular retinoic acid-binding proteins (CRABPs), 3) constitutive degradation of PML-RAR alpha, 4) point mutations in the ligand-binding domain of RAR alpha of PML-RAR alpha, 5) P-glycoprotein expression, 6) transcriptional repression by histone deacetylase activity, 7) isoforms of PML-RAR alpha, 8) persistent telomerase activity, and 9) expression of type II transglutaminase. In this review, we discuss the evidence provided in support of each mechanism, the mechanism's possible impact on the outcome of APL, and the newer approaches that are being employed to overcome ATRA resistance. PMID- 11501969 TI - Identification and characterization of androgen receptor associated coregulators in prostate cancer cells. AB - The androgen receptor (AR) is a member of the nuclear receptor (NR) superfamily that mediates the effects of androgens on target tissues. Over the last decade, it has become apparent that NRs require accessory factors for optimal activation of target gene expression. Numerous NR coregulators have been identified, with diverse structures and potential mechanisms of coregulation, creating an increasingly complicated picture of NR action. Due to the expanding complexity of the coregulator field, this review will focus on the AR ligand-binding domain (LBD) and N-terminal interacting proteins identified by our lab. The LBD interacting proteins ARA70, ARA55 and ARA54 were first characterized and ARA70 was found to have a relatively higher specificity for the AR in human prostate cancer DU145 cells. Characterization of the functional relationship between the AR and these coregulators indicated that ARA70 and ARA55 could enhance the androgenic effects of 17beta-estradiol (E2) and hydroxyflutamide (HF), an antiandrogen commonly used in the treatment of prostate cancer. ARA160, an AR N terminal interacting protein also known as TATA element modulatory factor (TMF), was subsequently shown to cooperate with ARA70 in enhancing AR activity. Another AR N-terminal interacting protein, ARA24, interacted with the poly-Q tract, a region within the N-terminus of the AR linked to Kennedy's disease (X-linked spinal and bulbar muscular atrophy). More recently, our lab has identified ARA267, a SET domain containing protein, and supervillin, an F-actin binding protein, as AR coregulators. Collectively, the data from these studies indicate that these coregulators are necessary for optimal AR transactivation. Interruption of the interaction between AR and these proteins may serve as a new therapeutic target in the treatment of prostate cancer. PMID- 11501970 TI - Transmembrane proteases as disease markers and targets for therapy. AB - Transmembrane proteases (i.e. membrane-associated proteases, ectoproteases) are present in a wide variety of tissues and cell types including endothelial, epithelial and hematopoietic cells. Natural and synthetic inhibitors have been characterized and have revealed that certain ectoenzymes are able to modulate bioactive peptide responses and to influence major biological events such as cell proliferation, survival and invasiveness. Dysregulated expression of some of them in human diseases triggers research on their role in pathophysiology, on their value as disease markers and as putative targets for therapy. PMID- 11501971 TI - Abnormally enhanced blood concentrations of vascular endothelial growth factor (VEGF) in metastatic cancer patients and their relation to circulating dendritic cells, IL-12 and endothelin-1. AB - Elevated VEGF blood concentrations have been proven to be associated with poor prognosis in human neoplasms. This finding is generally explained as a consequence of the potential angiogenic properties of VEGF itself. However, preliminary experimental studies suggest that VEGF, in addition to its angiogenic activity, may also play an immunosuppressant role by inhibiting dendritic cell (DC) maturation. The present study was performed to analyze blood levels of VEGF in cancer patients in relation to those of another potentially angiogenic tumor growth factor, endothelin-1 (ET-1), and to the absolute number of circulating immature and mature DC, and serum levels of the best known antitumor cytokine, IL 12. The study was performed in 100 healthy controls and in 80 solid tumor patients (colorectal cancer: 24; gastric cancer: 17; cancer of pancreas: 4; lung cancer: 13; breast cancer: 11; renal cell cancer: 6; gynecologic tumors: 5), 48 of whom showed distant organ metastases. In each patient, we have evaluated serum concentrations of VEGF-165, total VEGF, ET-1, IL-12 and the circulating number of immature (CD123+) and mature (CD11c+) DC. Mean serum levels of VEGF-165 were significantly higher in metastatic patients than in controls or in non-metastatic patients, whereas the total amounts of VEGF were not significantly higher. Moreover, it has been observed that patients with abnormally elevated blood concentrations of VEGF-165 showed significantly lower mean values of immature DC, mature DC and IL-12 and significantly higher mean levels of ET-1 than those with normal concentrations. This study, by confirming that advanced neoplastic disease may be associated with increased endogenous secretion of VEGF, seems to suggest that the association between high blood levels of VEGF and poor prognosis in cancer does not depend only on VEGF-induced stimulation of the neovascularization, but also on VEGF-related immunosuppression. PMID- 11501972 TI - Monoclonal antibody panels for acute leukemia and myelodysplastic syndrome diagnosis. Results of a co-operative quality control group. AB - The need for standardization criteria and result reproducibility in immunophenotyping hematological diseases has increased along with their clinical importance. Our group "Policentric Study Group on Immunological Markers", is composed of 40 laboratories. Its aim, over recent years, has been to find a standardized way of immunophenotypic analysis applicable to various hematological diseases. The objective of this study is to contribute to the debate concerning standardization of monoclonal antibody panels and immunophenotypic analysis procedures in acute leukemia (AL) and myelodysplastic syndrome (MDS), with the following targets: to improve interlaboratory reproducibility of the immunophenotyping data, and interpretative results; to study, with improved feasibility, correlation between immunophenotype and clinical or biological findings on a large number of AL and MDS cases; to verify the utility of the proposed monoclonal antibody panels for proper AL and MDS classification, and to detect minimal residual disease. In the field of AL and MDS our experience is based on about 1800 and 700 cases respectively analyzed over the last five years. Starting from these experiences and data of the literature we have elaborated the proposed panels of monoclonal antibodies and the methods of analysis. We have suggested a standardized immunophenotypic approach to study AL and MDS. In particular our work has focused on the gating strategy. This aims at drawing a gate of analysis having high purity and recovery, and on the choice of monoclonal antibody combinations for multiparametric analysis, particularly the normal antigen expression on each step of lineage differentiation or their clinically relevant aberrant expressions. A standardized criteria has become a necessary starting point in any kind of analytical process. In the field of acute leukemias and myelodysplastic syndromes the work of this polycentric group has focused on the pre-analytical and analytical steps to be taken in cytometric evaluation of hematological malignancies. The results obtained may contribute to reaching intra and inter-laboratory reproducibility. PMID- 11501973 TI - ACTH receptor distribution and modulation among murine mononuclear leukocyte populations. AB - Murine mononuclear leukocytes express adrenocorticotropin (ACTH) receptors that were recognized by a monospecific antiserum to the ACTH receptor on Y-1 adrenal cells. The antiserum was utilized in an immunofluorescence (IF) assay to characterize the distribution of ACTH receptors on resting murine mononuclear leukocyte populations. Forty-seven percent of spleen cells, 32% of lymph node cells, and 1% of thymocytes constitutively expressed ACTH receptors. Separation of lymphocytes into purified B cell and T cell populations, followed by IF analysis revealed that 47% of B cells and 23% of T cells possessed ACTH receptors. Helper T cells (CD4+ T cells) constituted the majority of ACTH receptor-positive T lymphocytes. Furthermore, 47% of resident peritoneal macrophages, purified by adherence to plastic, expressed ACTH receptors. The T lymphocyte mitogen, concanavalin A, interferon gamma, and ACTH enhanced ACTH receptor expression. The differential distribution of ACTH receptor-positive cells among specific leukocyte populations explains in part why differential cellular responses are observed and implies important regulatory functions for these receptors in the generation or regulation of immune responses. PMID- 11501974 TI - Stimulation of IL-12 secretion by GM-CSF in advanced cancer patients. AB - In addition to its efficacy in the treatment of chemotherapy-induced neutropenia, recent evidence would suggest that GM-CSF may have immunomodulatory effects on anticancer immunity. In particular, GM-CSF has been proven to promote dendritic cell maturation, with following potential stimulation of the anticancer cytokine, IL-12. Unfortunately, at present there are only few and controversial results on GM-CSF effects on IL-12 secretion in cancer patients. This preliminary study was performed to evaluate IL-12 response to an acute injection of GM-CSF in human neoplasms. The study included 20 advanced cancer patients, who received GM-CSF for chemotherapy-induced neutropenia. GM-CSF was injected at 3 micrograms/kg at 8.00 A.M., and venous blood samples were drawn before GM-CSF, and at 4, 8, 12 and 24 hours after its injection. Serum levels of IL-12 were measured by an enzyme immunoassay. High basal levels of IL-12 were seen in 8/20 patients. In patients with abnormally high pretreatment levels of IL-12, no significant change occurred in IL-12 mean serum concentration after GM-CSF administration. In contrast, patients with normal baseline levels of IL-12 showed a significant increase in IL 12 mean concentrations in response to GM-CSF, with a peak after 12 hours. This preliminary study seems to show that GM-CSF may acutely stimulate IL-12 secretion in cancer patients. Further studies are required to evaluate the effects of chronic GM-CSF administration, and the impact of GM-CSF-induced secretion of IL 12 on the efficacy of the immunotherapies of cancer with cytokines, such as IL-2. In any case, this study would justify further research in the emerging oncological applications of GM-CSF as an immunomodulatory agent on host anticancer defenses. PMID- 11501975 TI - Comparison of protease genotype and phenotype in HIV-1 infected patients exposed to more than one protease inhibitor. AB - Many patterns of mutations selected by HIV-1 protease inhibitors have been described, but in most cases isolates with these patterns have been obtained from pre-clinical studies or after failures of monotherapies. We compared genotype and phenotype in HIV-1 infected patients who have failed more than one PI-including regimen. Phenotypic resistance could arise also in the absence of specific primary mutations and in the presence of different substitutions among those known to confer resistance to ritonavir, indinavir or nelfinavir. The number of secondary mutations was significantly associated with phenotypic resistance for each protease inhibitor. Thus, more study of mutational patterns in heavily pretreated patients is warranted; in the mean time treatment choices might be optimized if phenotyping could integrate genotyping within this setting. PMID- 11501976 TI - First International Workshop on Human/SCID Mouse Models. PMID- 11501977 TI - Information resources for CD and related molecules. PMID- 11501978 TI - CD89 (Fc alphaRI). PMID- 11501980 TI - CD54 (ICAM-1). PMID- 11501979 TI - CD24. PMID- 11501981 TI - CD102 (ICAM-2). PMID- 11501982 TI - CD58 (LFA-3). PMID- 11501983 TI - CD104. PMID- 11501984 TI - New insights in medical mycology: focus on fungal infections. Introduction. PMID- 11501985 TI - In vitro antifungal susceptibility testing. AB - With the rising frequency of fungal infections, as well as increasing reports of resistance to antifungal agents, it is imperative that clinically applicable antifungal susceptibility testing be available. In 1997 the National Committee for Clinical Laboratory Standards published standard guidelines for antifungal susceptibility testing of Candida sp and Cryptococcus neoformans with amphotericin B, flucytosine, fluconazole, itraconazole, and ketoconazole. Although the methods are standard, they are time consuming, can be difficult to interpret, and are approved only for testing limited organisms and drugs. Modifications to the methods and alternative approaches have been proposed to make these tests more convenient and efficient, applicable to a greater number of species, and appropriate for performing in the clinical laboratory. PMID- 11501986 TI - Antifungal resistance among Candida species. AB - Isolation of Candida sp less susceptible to traditional therapies and recovery of increasingly resistant isolates during antifungal therapy are growing problems. It is important for clinicians to be aware of trends and mechanisms responsible for the expression of resistance. PMID- 11501987 TI - Antifungal pharmacodynamics: concentration-effect relationships in vitro and in vivo. AB - The pharmacodynamics of antifungal compounds involve relationships among drug concentrations, time, and antimicrobial effects in vitro and in vivo. Beyond better understanding of a drug's mode of action, characterization of these relationships has important implications for setting susceptibility breakpoints, establishing rational dosing regimens, and facilitating drug development. Important advances have been made in the experimental investigation of pharmacokinetics and pharmacodynamics of antifungal drugs; however, much remains to be learned about specific pathogens and specific sites of infection. Increased incorporation of pharmacokinetic and pharmacodynamic principles in experimental and clinical studies with antifungal agents is an important objective that will benefit the treatment and prophylaxis of life-threatening invasive fungal infections in immunocompromised patients. PMID- 11501988 TI - Rationale for combination antifungal therapy. AB - The relentless increase of invasive fungal infections and poor outcomes associated with available antifungal agents prompted the search for better therapeutic strategies. Combining antifungal drugs was recommended as a means to enhance efficacy in a variety of invasive infections including cryptococcosis, candidiasis, and aspergillosis. With the exception of cryptococcal meningitis, data from controlled clinical trials supporting such combinations are sparse. Moreover, little consensus exists regarding which combinations are synergistic or antagonistic in vitro and in vivo. Based on available data, several principles underlie these combinations. PMID- 11501989 TI - Investigational antifungal agents. AB - Several new antifungal agents, including novel compounds in familiar classes and entirely new classes targeting previously untapped mechanisms, are in various stages of the drug development process. Many new triazole antifungal agents are being studied, including voriconazole, posaconazole, and ravuconazole. The echinocandin antifungals, which represent a new class of antifungal agents, possess activity against a variety of fungal pathogens. The sodarin derivatives and nikkomycins are two additional classes of antifungals in early stages of development; future studies will determine their therapeutic usefulness. PMID- 11501990 TI - A clinical perspective for the management of invasive fungal infections: focus on IDSA guidelines. Infectious Diseases Society of America. AB - Invasive fungal infections, especially candidiasis and aspergillosis, are a major cause of morbidity and mortality. Many controversies surround the management of these infections. A critical overview of the recent Infectious Diseases Society of America practice guidelines is provided, as are comments on both the conundrums and future perspectives in medical mycology. PMID- 11501991 TI - Recognition of nucleotide G745 in 23 S ribosomal RNA by the rrmA methyltransferase. AB - Methylation of the N1 position of nucleotide G745 in hairpin 35 of Escherichia coli 23 S ribosomal RNA (rRNA) is mediated by the methyltransferase enzyme RrmA. Lack of G745 methylation results in reduced rates of protein synthesis and growth. Addition of recombinant plasmid-encoded rrmA to an rrmA-deficient strain remedies these defects. Recombinant RrmA was purified and shown to retain its activity and specificity for 23 S rRNA in vitro. The recombinant enzyme was used to define the structures in the rRNA that are necessary for the methyltransferase reaction. Progressive truncation of the rRNA substrate shows that structures in stem-loops 33, 34 and 35 are required for methylation by RrmA. Multiple contacts between nucleotides in these stem-loops and RrmA were confirmed in footprinting experiments. No other RrmA contact was evident elsewhere in the rRNA. The RrmA contact sites on the rRNA are inaccessible in ribosomal particles and, consistent with this, 50 S subunits or 70 S ribosomes are not substrates for RrmA methylation. RrmA resembles the homologous methyltransferase TlrB (specific for nucleotide G748) as well as the Erm methyltransferases (nucleotide A2058), in that all these enzymes methylate their target nucleotides only in the free RNA. After assembly of the 50 S subunit, nucleotides G745, G748 and A2058 come to lie in close proximity lining the peptide exit channel at the site where macrolide, lincosamide and streptogramin B antibiotics bind. PMID- 11501992 TI - Counterion association with native and denatured nucleic acids: an experimental approach. AB - The melting temperature of the poly(dA) . poly(dT) double helix is exquisitely sensitive to salt concentration, and the helix-to-coil transition is sharp. Modern calorimetric instrumentation allows this transition to be detected and characterized with high precision at extremely low duplex concentrations. We have taken advantage of these properties to show that this duplex can be used as a sensitive probe to detect and to characterize the influence of other solutes on solution properties. We demonstrate how the temperature associated with poly(dA) . poly(dT) melting can be used to define the change in bulk solution cation concentration imparted by the presence of other duplex and triplex solutes, in both their native and denatured states. We use this information to critically evaluate features of counterion condensation theory, as well as to illustrate "crosstalk" between different, non-contacting solute molecules. Specifically, we probe the melting of a synthetic homopolymer, poly(dA) . poly(dT), in the presence of excess genomic salmon sperm DNA, or in the presence of one of two synthetic RNA polymers (the poly(rA) . poly(rU) duplex or the poly(rU) . poly(rA) . poly(rU) triplex). We find that these additions cause a shift in the melting temperature of poly(dA) . poly(dT), which is proportional to the concentration of the added polymer and dependent on its conformational state (B versus A, native versus denatured, and triplex versus duplex). To a first approximation, the magnitude of the observed tm shift does not depend significantly on whether the added polymer is RNA or DNA, but it does depend on the number of strands making up the helix of the added polymer. We ascribe the observed changes in melting temperature of poly(dA) . poly(dT) to the increase in ionic strength of the bulk solution brought about by the presence of the added nucleic acid and its associated counterions. We refer to this communication between non-contacting biopolymers in solution as solvent-mediated crosstalk. By comparison with a known standard curve of tm versus log[Na+] for poly(dA) . poly(dT), we estimate the magnitude of the apparent change in ionic strength resulting from the presence of the bulk nucleic acid, and we compare these results with predictions from theory. We find that current theoretical considerations correctly predict the direction of the t(m) shift (the melting temperature increases), while overestimating its magnitude. Specifically, we observe an apparent increase in ionic strength equal to 5% of the concentration of the added duplex DNA or RNA (in mol phosphate), and an additional apparent increase of about 9.5 % of the nucleic acid concentration (mol phosphate) upon denaturation of the added DNA or RNA, yielding a total apparent increase of 14.5 %. For the poly(rU) . poly(rA) . poly(rU) triplex, the total apparent increase in ionic strength corresponds to about 13.6% of the amount of added triplex (moles phosphate). The effect we observe is due to coupled equilibria between the solute molecules mediated by modulations in cation concentration induced by the presence and/or the transition of one of the solute molecules. We note that our results are general, so one can use a different solute probe sensitive to proton binding to characterize subtle changes in solution pH induced by the presence of another solute in solution. We discuss some of the broader implications of these measurements/results in terms of nucleic acid melting in multicomponent systems, in terms of probing counterion environments, and in terms of potential regulatory mechanisms. PMID- 11501993 TI - Structural organisation of the head-to-tail interface of a bacterial virus. AB - In tailed icosahedral bacteriophages the connection between the 5-fold symmetric environment of the portal vertex in the capsid and the 6-fold symmetric phage tail is formed by a complex interface structure. The current study provides the detailed analysis of the assembly and structural organisation of such an interface within a phage having a long tail. The region of the interface assembled as part of the viral capsid (connector) was purified from DNA-filled capsids of the Bacillus subtilis bacteriophage SPP1. It is composed of oligomers of gp6, the SPP1 portal protein, of gp15, and of gp16. The SPP1 connector structure is formed by a mushroom-like portal protein whose cap faces the interior of the viral capsid in intact virions, an annular structure below the stem of the mushroom, and a second narrower annulus that is in direct contact with the helical tail extremity. The layered arrangement correlates to the stacking of gp6, gp15, and gp16 on top of the tail. The gp16 ring is exposed to the virion outside. During SPP1 morphogenesis, gp6 participates in the procapsid assembly reaction, an early step in the assembly pathway, while gp15 and gp16 bind to the capsid portal vertex after viral chromosome encapsidation. gp16 is processed during or after tail attachment to the connector region. The portal protein gp6 has 12-fold cyclical symmetry in the connector structure, whereas assembly-naive gp6 exhibits 13-fold symmetry. We propose that it is the interaction of gp6 with other viral morphogenetic proteins that drives its assembly into the 12-mer state. PMID- 11501994 TI - Investigation of the mechanism of domain closure in citrate synthase by molecular dynamics simulation. AB - Six, 2 ns molecular dynamics simulations have been performed on the homodimeric enzyme citrate synthase. In three, both monomers were started from the open, unliganded X-ray conformation. In the remaining three, both monomers started from a closed, liganded X-ray conformation, with the ligands removed. Projecting the motion from the simulations onto the experimental domain motion revealed that the free-energy profile is rather flat around the open conformation, with steep sides. The most closed conformations correspond to hinge-bending angles of 12-14 compared to the 20 degrees that occurs upon the binding of oxaloacetate. It is also found that the open, unliganded X-ray conformation is situated at the edge of the steep rise in free energy, although conformations that are about 5 degrees more open were sampled. A rigid-body essential dynamics analysis of the combined open trajectories has shown that domain motions in the direction of the closed X ray conformation are compatible with the natural domain motion of the unliganded protein, which has just two main degrees of freedom. The simulations starting from the closed conformation suggest a free-energy profile with a small barrier in going from the closed to open conformation. A combined essential dynamics and hinge-bending analysis of a trajectory that spontaneously converts from the closed to open state shows an almost exact correspondence to the experimental transition that occurs upon ligand binding. The simulations support the conclusion from an earlier analysis of the experimental transition that the beta hairpin acts as a mechanical hinge by attaching the small domain to the large domain through a conserved main-chain hydrogen bond and salt-bridges, and allowing rotation to occur via its two flexible termini. The results point to a mechanism of domain closure in citrate synthase that has analogy to the process of closing a door. PMID- 11501995 TI - Type II protein secretion in gram-negative pathogenic bacteria: the study of the structure/secretion relationships of the cellulase Cel5 (formerly EGZ) from Erwinia chrysanthemi. AB - Erwinia chrysanthemi, a Gram-negative plant pathogen, secretes the cellulase Cel5 (formerly EGZ) via the type II secretion pathway (referred to as Out). Cel5 is composed of two domains, a large N-terminal catalytic domain (390 amino acid residues) and a small C-terminal cellulose-binding domain (62 amino acid residues) separated by a linker region. A combination of mutagenesis and structural analysis permitted us to investigate the structure/secretion relationships with respect to the catalytic domain of Cel5. The 3D structure of the catalytic domain was solved by molecular replacement at 2.3 A resolution. Cel5 exhibits the (beta/alpha)8 structural fold and two extra-barrel features. Our previous genetic study based upon tRNA-mediated suppression allowed us to predict positions of importance in the molecule in relation to structure and catalysis. Remarkably, all of the predictions proved to be correct when compared with the present structural information. Mutations of Arg57, which is located at the heart of the catalytic domain, allowed us to test the consequences of structural modifications on the secretion efficiency. The results revealed that secretability imposes remarkably strong constraints upon folding. In particular, an Arg-to-His mutation yielded a species that folded to a stable conformation close to, but distinct from the wild-type, which however was not secretable. We discuss the relationships between folding of a protein in the periplasm, en route to the cell exterior, and presentation of secretion information. We propose that different solutions have been selected for type II secreted exoproteins in order to meet the constraints imposed by their interaction with their respective secretion machineries. We propose that evolutionary pressure has led to the adaptation of different secretion motifs for different type II exoproteins. PMID- 11501996 TI - Folate-binding triggers the activation of folylpolyglutamate synthetase. AB - Folic acid is an essential vitamin for normal cell growth, primarily through its central role in one-carbon metabolism. Folate analogs (antifolates) are targeted at the same reactions and are widely used as therapeutic drugs for cancer and bacterial infections. Effective retention of folates in cells and the efficacy of antifolate drugs both depend upon the addition of a polyglutamate tail to the folate or antifolate molecule by the enzyme folylpolyglutamate synthetase (FPGS). The reaction mechanism involves the ATP-dependent activation of the free carboxylate group on the folate molecule to give an acyl phosphate intermediate, followed by attack by the incoming L-glutamate substrate. FPGS shares a number of structural and mechanistic details with the bacterial cell wall ligases MurD, MurE and MurF, and these enzymes, along with FPGS, form a subfamily of the ADP forming amide bond ligase family. High-resolution crystallographic analyses of binary and ternary complexes of Lactobacillus casei FPGS reveal that binding of the first substrate (ATP) is not sufficient to generate an active enzyme. However, binding of folate as the second substrate triggers a large conformational change that activates FPGS and allows the enzyme to adopt a form that is then able to bind the third substrate, L-glutamate, and effect the addition of a polyglutamate tail to the folate. PMID- 11501997 TI - Crystal structure of alkaline cellulase K: insight into the alkaline adaptation of an industrial enzyme. AB - The crystal structure of the catalytic domain of alkaline cellulase K was determined at 1.9 A resolution. Because of the most alkaliphilic nature and it's highest activity at pH 9.5, it is used commercially in laundry detergents. An analysis of the structural bases of the alkaliphilic character of the enzyme suggested a mechanism similar to that previously proposed for alkaline proteases, that is, an increase in the number of Arg, His, and Gln residues, and a decrease in Asp and Lys residues. Some ion pairs were formed by the gained Arg residues, which is similar to what has been found in the alkaline proteases. Lys-Asp ion pairs are disfavored and partly replaced with Arg-Asp ion pairs. The alkaline adaptation appeared to be a remodeling of ion pairs so that the charge balance is kept in the high pH range. PMID- 11501998 TI - Solution structure of the catalytic domain of gammadelta resolvase. Implications for the mechanism of catalysis. AB - The site-specific DNA recombinase, gammadelta resolvase, from Escherichia coli catalyzes recombination of res site-containing plasmid DNA to two catenated circular DNA products. The catalytic domain (residues 1-105), lacking a C terminal dimerization interface, has been constructed and the NMR solution structure of the monomer determined. The RMSD of the NMR conformers for residues 2-92 excluding residues 37-45 and 64-73 is 0.41 A for backbone atoms and 0.88 A for all heavy atoms. The NMR solution structure of the monomeric catalytic domain (residues 1-105) was found to be formed by a four-stranded parallel beta-sheet surrounded by three helices. The catalytic domain (residues 1-105), deficient in the C-terminal dimerization domain, was monomeric at high salt concentration, but displayed unexpected dimerization at lower ionic strength. The unique solution dimerization interface at low ionic strength was mapped by NMR. With respect to previous crystal structures of the dimeric catalytic domain (residues 1-140), differences in the average conformation of active-site residues were found at loop 1 containing the catalytic S10 nucleophile, the beta1 strand containing R8, and at loop 3 containing D67, R68 and R71, which are required for catalysis. The active-site loops display high-frequency and conformational backbone dynamics and are less well defined than the secondary structures. In the solution structure, the D67 side-chain is proximal to the S10 side-chain making the D67 carboxylate group a candidate for activation of S10 through general base catalysis. Four conserved Arg residues can function in the activation of the phosphodiester for nucleophilic attack by the S10 hydroxyl group. A mechanism for covalent catalysis by this class of recombinases is proposed that may be related to dimer interface dissociation. PMID- 11501999 TI - Solution structure of the pseudoknot of SRV-1 RNA, involved in ribosomal frameshifting. AB - RNA pseudoknots play important roles in many biological processes. In the simian retrovirus type-1 (SRV-1) a pseudoknot together with a heptanucleotide slippery sequence are responsible for programmed ribosomal frameshifting, a translational recoding mechanism used to control expression of the Gag-Pol polyprotein from overlapping gag and pol open reading frames. Here we present the three dimensional structure of the SRV-1 pseudoknot determined by NMR. The structure has a classical H-type fold and forms a triple helix by interactions between loop 2 and the minor groove of stem 1 involving base-base and base-sugar interactions and a ribose zipper motif, not identified in pseudoknots so far. Further stabilization is provided by a stack of five adenine bases and a uracil in loop 2, enforcing a cytidine to bulge. The two stems of the pseudoknot stack upon each other, demonstrating that a pseudoknot without an intercalated base at the junction can induce efficient frameshifting. Results of mutagenesis data are explained in context with the present three-dimensional structure. The two base pairs at the junction of stem 1 and 2 have a helical twist of approximately 49 degrees, allowing proper alignment and close approach of the three different strands at the junction. In addition to the overwound junction the structure is somewhat kinked between stem 1 and 2, assisting the single adenosine in spanning the major groove of stem 2. Geometrical models are presented that reveal the importance of the magnitude of the helical twist at the junction in determining the overall architecture of classical pseudoknots, in particular related to the opening of the minor groove of stem 1 and the orientation of stem 2, which determines the number of loop 1 nucleotides that span its major groove. PMID- 11502000 TI - Real time kinetic studies of the interaction between folded antisense and target RNAs using surface plasmon resonance. AB - Antisense RNAs interact with their complementary target RNAs as folded structures. The formation of early binding intermediates is the most important step in determining the overall rates of stable complex formation in vitro and the efficiency of control in vivo. In the case of CopA and CopT (antisense/target RNA pair of plasmid R1), recent studies have identified a four-way junction structure as the major binding intermediate. Previously, the kinetics of antisense/target RNA interaction was studied by indirect methods. Here we have used surface plasmon resonance to follow the binding of CopI (a truncated variant of CopA) to CopT in real time. A protocol was developed that permitted the determination of association and dissociation rate constants for wild-type and mutant CopI-CopT pairs. The K(D)-values calculated from these rate constants were in good agreement with the results obtained by indirect methods. In comparison to earlier model studies of interactions between simple complementary nucleic acids, we observe a different temperature dependence for dissociation rate constants. This may be indicative of the complexity of the steps required for interacting folded RNAs; intramolecular structure competes with intermolecular helix progression during complex formation. The association rate constants were not significantly dependent on temperature. The analysis presented shows that the stability of a kissing complex is not the primary determinant of the rate of stable CopA/CopT complex formation. PMID- 11502001 TI - Defining linear segments in protein structure. AB - The analysis of protein structure using secondary structure line segments has been widely used in many structure analysis and prediction methods over the past 20 years. Its use in methods that compare protein structures at this level of representation is becoming more important as an increasing number of protein structures become determined through structural genomic programmes. The standard method used to define line segments is to fit an axis through each secondary structure element. This approach has difficulties, however, both with inconsistent definitions of secondary structure and the problem of fitting a single straight line to a bent structure. The procedure described here avoids these problems by finding a set of line segments independently of any external secondary structure definition. This allows the segments to be used as a novel basis for secondary structure definition by taking the average rise/residue along each axis to characterise the segment. This practice has the advantage that secondary structures are described by a single (continuous) value that is not restricted to the conventional classes of alpha-helix, 310 and beta-strand. This latter property allows structures without "classic" secondary structures to be encoded as line segments that can be used in comparison algorithms. When compared over a large number of pairs of homologous proteins, the current method was found to be slightly more consistent than a widely used method based on hydrogen bonds. PMID- 11502002 TI - In silico structural and functional analysis of the human cytomegalovirus (HHV5) genome. AB - The open reading frames of human cytomegalovirus (human herpesvirus-5, HHV5) encode some 213 unique proteins with mostly unknown functions. Using the threading program, ProCeryon, we calculated possible matches between the amino acid sequences of these proteins and the Protein Data Bank library of three dimensional structures. Thirty-six proteins were fully identified in terms of their structure and, often, function; 65 proteins were recognized as members of narrow structural/functional families (e.g. DNA-binding factors, cytokines, enzymes, signaling particles, cell surface receptors etc.); and 87 proteins were assigned to broad structural classes (e.g. all-beta, 3-layer-alphabetaalpha, multidomain, etc.). Genes encoding proteins with similar folds, or containing identical structural traits (extreme sequence length, runs of unstructured (Pro and/or Gly-rich) residues, transmembrane segments, etc.) often formed tandem clusters throughout the genome. In the course of this work, benchmarks on about 20 known folds were used to optimize adjustable parameters of threading calculations, i.e. gap penalty weights used in sequence/structure alignments; new scores obtained as simple combinations of existing scoring functions; and number of threading runs conducive to meaningful results. An introduction of summed, per residue-normalized scores has been essential for discovery of subdomains (EGF like, SH2, SH3) in longer protein sequences, such as the eight "open sandwich" cytokine domains, 60-70 amino acids long and having the 3beta1alpha fold with one or two disulfide bridges, present in otherwise unrelated proteins. PMID- 11502003 TI - High-resolution structure of HLA-A*0201 in complex with a tumour-specific antigenic peptide encoded by the MAGE-A4 gene. AB - The heterotrimeric complex of the human major histocompatibity complex (MHC) molecule HLA-A*0201, beta2-microglobulin and the decameric peptide GVYDGREHTV derived from the melanoma antigen (MAGE-A4 protein has been determined by X-ray crystallography at 1.4 A resolution. MAGE-A4 belongs to a family of genes that are specifically expressed in a variety of tumours. MAGE-A4-derived peptides are presented by MHC molecules at the cell surface to cytotoxic T-lymphocytes. As the HLA-A*0201:MAGE-A4 complex occurs only on tumour cells, it is considered to be an appropriate target for immunotherapy. The structure presented here reveals potential epitopes specific to the complex and indicates which peptide residues could be recognised by T-cell receptors. In addition, as the structure could be refined anisotropically, it was possible to describe the movements of the bound peptide in more detail. PMID- 11502004 TI - The osmophobic effect: natural selection of a thermodynamic force in protein folding. AB - Intracellular organic osmolytes are present in certain organisms adapted to harsh environments and these osmolytes protect intracellular macromolecules against the denaturing environmental stress. In natural selection of organic osmolytes as protein stabilizers, it appears that the osmolyte property selected for is the unfavorable interaction between the osmolyte and the peptide backbone, a solvophobic thermodynamic force that we call the osmophobic effect. Because the peptide backbone is highly exposed to osmolyte in the denatured state, the osmophobic effect preferentially raises the free energy of the denatured state, shifting the equilibrium in favor of the native state. By focusing the solvophobic force on the denatured state, the native state is left free to function relatively unfettered by the presence of osmolyte. The osmophobic effect is a newly uncovered thermodynamic force in nature that complements the well recognized hydrophobic interactions, hydrogen bonding, electrostatic and dispersion forces that drive protein folding. In organisms whose survival depends on the intracellular presence of osmolytes that can counteract denaturing stresses, the osmophobic effect is as fundamental to protein folding as these well-recognized forces. PMID- 11502005 TI - Biogenesis of the dicarboxylate carrier (DIC): translocation across the mitochondrial outer membrane and subsequent release from the TOM channel are membrane potential-independent. AB - The mitochondrial inner membrane of Saccharomyces cerevisiae contains a group of homologous carrier proteins that mediate the exchange of several metabolites. The members of this protein family are synthesized in the cytosol and reach their final topology after translocation across the mitochondrial outer membrane. Using the ADP/ATP carrier (AAC) as a model protein, previous studies have established four distinct steps of the import pathway (stages I-IV). In the absence of the mitochondrial membrane potential (deltapsi), the AAC accumulates at the inner surface of the outer membrane (stage IIIa) and remains bound to the outer membrane import channel. Only in the presence of the membrane potential, can a complex of small Tim proteins mediate transfer of the AAC to the inner membrane. In this study, we characterized the import pathway of the dicarboxylate carrier (DIC). Different from the AAC, the DIC showed complete deltapsi-independent translocation across the outer membrane, release from the import pore, and mainly accumulated in a soluble state in the intermembrane space, thus defining a new translocation intermediate (stage III*). The DIC should be a suitable model protein for the characterization of deltapsi-independent functions of the intermembrane space Tim proteins. PMID- 11502006 TI - Structural conversion between open and closed forms of radixin: low-angle shadowing electron microscopy. AB - The function of ERM (ezrin/radixin/moesin) proteins as general cross-linkers between actin filaments and plasma membranes is regulated downstream of Rho, through the transition between active and inactive forms. To directly examine the conformational change between the active and inactive forms of ERM proteins, we applied low-angle rotary-shadowing electron microscopy to the radixin molecules, wild-type, T564A-non-phosphorylated-type, and T564E-phosphorylated-type, since most of the active forms are reportedly stabilized in cells by the C-terminal threonine phosphorylation. As a result, the T564A- and wild-type radixin molecules yielded the globular closed forms, approximately 8-14 nm in diameter, with some striations on their surfaces. In contrast, the T564E-radixin molecules tended to take elongated open forms, in which two globular structures measuring approximately 8 nm and approximately 5 nm in diameter were associated with both ends of the filamentous structures. The filamentous structure took either a approximately 20-25 nm-long straight course or a folded course. Taken together with the biochemical and the crystal structural results obtained to date, the closed and open forms represent the inactive and active forms of radixin as cross linkers between actin filaments and plasma membranes. PMID- 11502007 TI - Mechanism of Tet repressor induction by tetracyclines: length compensates for sequence in the alpha8-alpha9 loop. AB - Natural Tet repressor (TetR) variants are alpha-helical proteins bearing a large loop between helices 8 and 9, which is variable in sequence and length. We have deleted this loop consisting of 14 amino acid residues in TetR(D) and rebuilt it stepwise with up to 42 alanine residues. All except the mutant with the longest alanine loop show wild-type repression, but none is inducible with tetracycline. This demonstrates the importance of the alpha8-alpha9 loop and its amino acid sequence for induction. The induction efficiencies increase with loop length, when the more tightly binding inducer anhydrotetracycline is used. The largest increase of inducibility was observed for TetR mutants with loop lengths between eight and 17 alanine residues. Since loop residues Asp/Glu157 and Arg158 are conserved in the natural TetR sequence variants, we constructed a mutant in which all other residues of the loop were replaced by alanine. This mutant exhibits increased anhydrotetracycline induction compared to the corresponding alanine variant. Thus, these residues are important for induction. Binding constants for the anhydrotetracycline-TetR interaction are below the detection level of 10(5) M(-1) for the mutant with a loop of two alanine residues and increase sharply until a loop size of ten residues is reached. TetR variants with longer loops have similar anhydrotetracycline-binding constants, ranging between 2.6 x 10(9) M(-1) and 8.0 x 10(9) M(-1), about 500-fold lower than wild-type TetR. The increase of the affinity occurs at shorter loop lengths than that of inducibility. We conclude that the induction defect of the polyalanine variants arises from two increments: (i) the loop must have a minimal length-to allow efficient inducer binding; (ii) the loop must structurally participate in the conformational change associated with induction. PMID- 11502008 TI - A sequence required for -1 ribosomal frameshifting located four kilobases downstream of the frameshift site. AB - Programmed ribosomal frameshifting allows one mRNA to encode regulate expression of, multiple open reading frames (ORFs). The polymerase encoded by ORF 2 of Barley yellow dwarf virus (BYDV) is expressed via minus one (-1) frameshifting from the overlapping ORF 1. Previously, this appeared to be mediated by a 116 nt RNA sequence that contains canonical -1 frameshift signals including a shifty heptanucleotide followed by a highly structured region. However, unlike known -1 frameshift signals, the reporter system required the zero frame stop codon and did not require a consensus shifty site for expression of the -1 ORF. In contrast, full-length viral RNA required a functional shifty site for frameshifting in wheat germ extract, while the stop codon was not required. Increasing translation initiation efficiency by addition of a 5' cap on the naturally uncapped viral RNA, decreased the frameshift rate. Unlike any other known RNA, a region four kilobases downstream of the frameshift site was required for frameshifting. This included an essential 55 base tract followed by a 179 base tract that contributed to full frameshifting. The effects of most mutations on frameshifting correlated with the ability of viral RNA to replicate in oat protoplasts, indicating that the wheat germ extract accurately reflected control of BYDV RNA translation in the infected cell. However, the overall frameshift rate appeared to be higher in infected cells, based on immunodetection of viral proteins. These findings show that use of short recoding sequences out of context in reporter constructs may overlook distant signals. Most importantly, the remarkably long-distance interaction reported here suggests the presence of a novel structure that can facilitate ribosomal frameshifting. PMID- 11502009 TI - The financial impact of health promotion and disease prevention programs--why is it so hard to prove value? PMID- 11502010 TI - Conceptual framework, critical questions, and practical challenges in conducting research on the financial impact of worksite health promotion. AB - A conceptual framework to guide research on the financial impact of health promotion programs is described. Important questions that must be answered within this framework are discussed and brief summaries of the research addressing these questions are summarized. Key issues and challenges that are encountered in designing and implementing financial outcome research are reviewed. PMID- 11502011 TI - Getting closer to the truth: overcoming research challenges when estimating the financial impact of worksite health promotion programs. AB - The authors describe the most important methodological challenges often encountered in conducting research and evaluation on the financial impact of health promotion. These include selection bias, skewed data, small sample size, metrics. They discuss when these problems can and cannot be overcome and suggest how some of these problems can be overcome through a creating an appropriate framework for the study, and using state of the art statistical methods. PMID- 11502012 TI - Financial impact of health promotion programs: a comprehensive review of the literature. AB - PURPOSE: The purpose of this review is to summarize the literature on the ability of health promotion programs to reduce employee-related health care expenditures and absenteeism. SEARCH PROCESS: Using key words in a literature-searching program, a comprehensive search was conducted on the following databases: MEDLINE, Embase, HealthSTAR. SPORTDiscus, PsycINFO, SciSearch, ERIC, and ABI Inform. STUDY INCLUSION AND EXCLUSION CRITERIA: All data-based studies that appeared in peer reviewed journals in the English language. Theses, dissertations, or presentation abstracts that were not published in peer reviewed journals were excluded. The initial search identified 196 studies, but only 72 met the inclusion criteria and were included in the review. DATA EXTRACTION METHODS: Summary tables were created that include design classification, subject size, results, and other key information for each study. DATA SYNTHESIS: Both the nature of the findings and the overall quality of the literature were evaluated in an attempt to answer two questions: Do individuals or populations with high health risks have worse financial outcomes than individuals or populations with low health risks? Do health promotion programs improve financial outcomes? MAJOR CONCLUSIONS: There are good correlational data to suggest that high levels of stress, excessive body weight, and multiple risk factors are associated with increased health care costs and illness-related absenteeism. The associations between seat belt use, cholesterol, diet, hypertension, and alcohol abuse and absenteeism and health care expenditures are either mixed or unknown. Health promotion programs are associated with lower levels of absenteeism and health care costs, and fitness programs are associated with reduced health care costs. PMID- 11502013 TI - The financial impact of smoking on health-related costs: a review of the literature. AB - PURPOSE: To summarize our knowledge of the economic impact of smoking on health related outcomes and to discuss the quality of this evidence. The potential effect of smoking-related health promotion programs on reducing costs is discussed. DATA SOURCES: Studies reviewed were cited in recently published reviews, identified through database searches of MEDLINE/HealthSTAR, PsychINFO, and ABI/Inform or obtained from reference lists of other studies. STUDY INCLUSION AND EXCLUSION CRITERIA: Included are studies of the health care and other costs of smoking published in English in peer-reviewed journals during the last 20 years. Studies of annual and lifetime costs are included as are studies of employer costs. Forty-nine studies were reviewed. DATA EXTRACTION METHODS: Data extracted were total costs, medical care and other direct costs, lost productivity, and mortality costs. Underlying methodologies were described and compared. DATA SYNTHESIS: The annual cost of smoking is between 6 and 14% of personal health expenditures. Estimates have increased over time, reflecting better and more comprehensive methodologies. Smoking appears to result in increased lifetime costs, although some studies havefound contrary results. In the workplace, smokers incur greater medical costs and more lost productivity than nonsmokers. They also impose costs on their nonsmoking co-workers. MAJOR CONCLUSIONS: The overwhelming body of evidence in the literature asserts that smoking imposes costs on an annual basis, that it leads to increased medical costs over the life span, and that many of these costs are borne by employers. Although the methods for studying this issue have evolved over time and are presently quite comprehensive, future research is needed to further substantiate the findings and develop further refinements. PMID- 11502014 TI - Shining lights: studies that have most influenced the understanding of health promotion's financial impact. AB - PURPOSE: To examine the literature from the past 20 years and identify those studies that support the economic merit of health promotion. DATA SOURCE: A panel of experts was used to identify the top studies supporting the purpose of this article. STUDY INCLUSION AND EXCLUSION CRITERIA: Studies were chosen based on the following criteria: the study (1) examined the relationship between health risks and financial outcomes, or health promotion programs and financial outcomes; (2) provided strong and compelling financial data supporting the worth of health promotion; (3) had a high-quality methodology; (4) answered an important question or replicated important findings with superior methodology; and (5) represented U.S.-based initiatives published since 1980. After initially nominating a group of studies for consideration, panelists rated each on a scale from 1 to 3 representing their opinion of importance. Studies rating the highest were included for this discussion. DATA EXTRACTION METHODS: Studies were analyzed by population characteristics, design, statistical tests, limitations, and results. This information was summarized for each identified article. MAJOR CONCLUSIONS: A relationship between modifiable health risk factors and health care costs is supported by research. Health promotion interventions appear to provide positive financial returns, most notably for health care costs and absenteeism reduction. Private sector initiatives seem to be driving economic-based research. Overall, health promotion shows promising results for providing financial advantages for its sponsors; however, if this discipline is to show its true worth, considerable funding is needed from government or philanthropic sources to cover the substantial costs of quality research. PMID- 11502015 TI - Emerging research: a view from one research center. AB - The Health Management Research Center at the University of Michigan has assembled a database on health risks, medical care costs, an in some cases, productivity measures for over 2,000,000 individuals. For employees of its corporate consortium members, the database contains seven to eighteen years of data. Working with this data, the research team has observed a number of emerging trends. These trends have been stable in this data set for a number of years, but some of them are yet to be subjected to rigorous external peer review. The trends are summarized below. 1) Annual participation rates of 20% to 30% in Health Risk Appraisal are typical; over 10 years, 80% participate at least once, 60% at least twice and 40% at least three times. 2) Among the employers in the data base, excess risk factors account for 21% to 31% of medical care costs, with a mean of 25%. 3) Medical care costs increase as the number risk factors and age increase. As risk factors increase, medical costs increase; as risk factors decrease, medical care costs decrease. The mean cost increase per risk factor increased ($350) may be more than double the mean cost decrease per risk factor decreased ($150). 4) Cost savings greatest among those who participate in programs multiple times. 5) Absenteeism seems to be higher and other measures of productivity lower for those with health risk factors. 6) Programs designed to keep healthy people healthy in addition to reducing the risks of those with multiple risks will probably provide the greatest return to the employers. 7) Best results may be achieved by focusing efforts on employees who have clusters of risk factors associated with low perceived health status. 8) A corporate wellness score which combines risk factor levels and participation rates may provide a "corporate wellness score" which can be used to compare health status across employer. 9) Increased use of longitudinal data sets, fuzzy cut points for data categories and data mining techniques may allow breakthroughs in future analysis efforts. PMID- 11502016 TI - Expanding health insurance coverage for smoking cessation treatments: experience of the Pacific Business Group on Health. AB - The business case for health insurance coverage of smoking cessation treatments by employers is a strong one. Smoking is one of the nation's costliest health problems, in both human and financial terms. The science behind smoking cessation treatment and promotion of treatment is strong; the cost effectiveness of smoking cessation treatment is among the highest in all of medicine, the time required before a positive return on investment is reasonable for employers, and the short term costs of treatments are well estimated and manageable for health plans and employers. Armed with this business case, the PBGH Negotiating Alliance has expanded health insurance to include pharmacotherapy, over the counter or by prescription, and behavioral interventions. Because PBGH has been a national leader, we hope that other employers, employer coalitions, and public purchasers will follow their lead. The potential health effect of even small reductions in smoking are striking, and unlike other chronic illnesses, nicotine addiction is curable, at both individual and societal levels. Thus, if employers make the investment in smoking cessation and other tobacco control today, they face the real possibility that the need for such outlays could decrease in the future. PMID- 11502017 TI - A corporate perspective: reflections from the economic buyer of health promotion programs. PMID- 11502018 TI - The Glaxo Wellcome health promotion program: the contract for health and wellness. PMID- 11502019 TI - Prevention: is there value for corporate purchasers? PMID- 11502020 TI - A corporate perspective from Lucent Technologies: reflections from the economic buyer of health promotion programs--what really matters to me and my boss? PMID- 11502021 TI - An optimistic view from the corporate perspective. PMID- 11502022 TI - Johnson & Johnson LIVE FOR LIFE Program: now and then. PMID- 11502023 TI - A corporate perspective on health promotion: reflections and advice from Chevron. AB - In summary, our program's focus on helping-management meet their business needs has been the most important component of our survival. We have used a conceptual philosophy that involves health change on multiple levels: individual, environmental, and cultural. We have demonstrated results, but most have not been financial. Our preference would be to provide services that our management values and at the same time demonstrate that our services deliver a tangible financial return. Our difficulty in doing so has been the lack of an integrated database and a small budget. We have supported large-scale studies as a participant, both in the HERO database' and in studies conducted by the MEDSTAT group (in progress). We will continue to participate in such activities to further the science of health promotion. Despite the success of our programs at Chevron, we continue to have to defend ourselves against those who question the value of health promotion. This is particularly true during cost reduction initiatives that focus on cutting everything that is not required for the business. Health promotion is not a necessity in corporate America's viewpoint. As with most prevention programs, health promotion must prove itself to be taken seriously. Getting management's continued support is a combination of adding value to their business, doing so cost effectively, measuring outcomes and communicating them, and finally, impacting management in a way that they experience the positive impact of wellness on a very personal level. PMID- 11502024 TI - When the health promotion program does not adapt to changing business priorities: the experience at Southern California Edison. PMID- 11502025 TI - The 3M corporate experience: health as a business strategy. PMID- 11502026 TI - Strategy, not return on investment, drives health promotion at Applied Materials, Inc.. PMID- 11502027 TI - An overview of two worksite health promotion programs--the quest for qualitative and quantitative results: what really matters? PMID- 11502028 TI - Financial impact of health promotion: we need to know much more, but we know enough to act. PMID- 11502029 TI - A government perspective: if there is so much proof, why is Medicare not rapidly adopting health promotion and disease prevention programs? PMID- 11502030 TI - Why health promotion lags knowledge about healthful behavior. PMID- 11502031 TI - Does proof matter? why strong evidence sometimes yields weak action. PMID- 11502032 TI - Oral care for patients with cancer: a review of the literature. AB - Mouth care is an integral part of nursing practice. However, it has become a ritualistic and banal activity, a topic of conflicting advice and subjective conclusions from sporadic research. Rarely do experts teach it, and it frequently is delegated to the most junior members of the nursing staff. Many cancer treatments result in unavoidable oral toxicity, and the significant prevalence of oral complications in oncology makes mouth care a particular priority for cancer nurses. The confusion and conflict that surrounds best nursing practice in relation to delivering appropriate mouth care should be redressed. This review article details pertinent research to date regarding oral care, with an aim to clarify intervention options, and to identify and promote best nursing practice. PMID- 11502033 TI - Quality of life experienced by patients receiving radiation treatment for cancers of the head and neck. AB - The purpose of this descriptive study was to explore the quality of life for 58 outpatients during and after a course of radiation treatment for cancers of the head and neck. Data were collected at three time points: T1 (first week of treatment), T2 (last week of treatment), and T3 (1 month after treatment). Patients completed two measures that assessed physical, emotional, functional, and social aspects of well-being: the Functional Assessment of Cancer Therapy: Head and Neck (FACT-H&N) and the Hospital Anxiety and Depression Scale (HADS). Repeated measures multivariate analysis of variance and post hoc t tests were performed to assess changes in quality of life over the three time points. Results indicated overall increased levels of physical and functional symptoms, head and neck specific concerns, and depression between T1 and T2. However, except for depression, there was some improvement between scores on each of these measures of physical and functional well-being between T2 and T3, although this improvement was not to the pretreatment level. The FACT subscales assessing social and emotional well-being, and the HADS subscale showed no significant changes across time. Implications of this study relate to the ongoing need for interventions to assist patients once they have completed the radiation treatment course, and to improved assessment in some areas of emotional distress. PMID- 11502034 TI - Coping with radiation therapy: strategies used by women with breast cancer. AB - The purpose of this qualitative study was to describe coping strategies used by women with breast cancer during and after radiation therapy. Content analysis was used to structure the statements provided by the patients in the Wheel Questionnaire. The patients used several problem- and emotion-focused strategies to cope with the treatment, and the strategies changed over time. Family and friends had a positive impact on the coping process for the three points of measurement: before treatment started, on completion of treatment, and 3 months after treatment. Contact with colleagues at work also provided the women with a sense of normalcy. The women used a broad spectrum of own activities to aid recovery, instead of leaving their treatment solely to medicine. The women also found that a hopeful and optimistic attitude was a helpful strategy for coping with the treatment. PMID- 11502035 TI - Psychosocial stages and quality of life of women with breast cancer. AB - Certain issues are universal for all women with breast cancer, irrespective of age, ethnic group, or stage of disease. Yet, along with common concerns, experiences, and anxieties, each woman may encounter a unique set of problems. Ultimately, each woman's adaptation and choices will be strongly influenced by her personal history, her psychosocial stage, and her life-cycle concerns. Changes in the criteria norms for the psychosocial stages of women's lives and their subsequent influence on quality of life are issues that have substantial implications for nursing and other healthcare professions. Younger and older women have different needs, concerns, and quality of life issues in a context of psychosocial life stages that have changed significantly across the post-World War II generations. The experience of breast cancer is perceived differently by women of distinct psychosocial life stages. At each critical life stage, the unique emerging problems require specific psychosocial supports that can reduce or avert the ensuing emotional distress. The planning and implementation of care must be tailored to address the differences demonstrated by age and psychosocial life stage, and to enhance quality of life outcomes for survivors of breast cancer, both young and old. PMID- 11502036 TI - The aftermath of breast cancer: an altered sexual self. AB - The literature lacks descriptions of the effect that breast cancer treatments have on sexuality from the perspective of women who have lived through this experience. The purpose of this study was to describe aspects of sexuality that were important to women after breast cancer treatment. Participants in this study were 18 white women with breast cancer between the ages of 35 and 68 years (mean, 50.5 years). Time since diagnosis ranged from 6 months to more than 10 years. Primary surgical treatment for seven women (39%) had been lumpectomy, and for 11 women (61%) mastectomy. Most of the participants also received adjuvant chemotherapy. This qualitative descriptive study used grounded theory methods. Two primary categories (losses and influencing pieces) emerged from the data analysis. The losses category had four components: missing parts, loss of bleeding-becoming old, loss of sexual sensations, and loss of womanhood. The influencing pieces were relationships and information control. The core concept identified was an altered sexual self. Central to the task of adjustment to living with breast cancer was coming to terms with a new sexual self that emerged after treatment. Women who sought information about the sexual side effects of cancer treatment, and who had strong intimate relationships appeared to experience a more successful adjustment. PMID- 11502037 TI - The Human Genome Project: an update. AB - The mapping, sequencing, and analysis of the human genome that has occurred during the last decade through the Human Genome Project are providing fundamental advances for basic science and medicine. Genomic information is providing insights into causes of, susceptibility to, and protection from cancer and a host of other diseases. Already, information generated by the Human Genome Project has been incorporated into the care of cancer patients. Perhaps more so than other types of medical information, genetic knowledge can have profound implications for individuals, families, and society. As a result, nursing professionals in clinical and academic settings are being called upon to identify and deliberate medical, social, ethical, and legal issues stemming from Human Genome Project advancements. The purpose of this article is to review the goals and implications of the Human Genome Project to further prepare cancer nurses to actively participate in the deliberations, research, and clinical activities evolving from the Human Genome Project. PMID- 11502038 TI - Patterns of advanced cancer pain in home hospice patients. AB - The purpose of this descriptive study was to provide a description of advanced cancer pain in home hospice subjects. The McGill Pain Questionnaire, Parts 1 and 2, measured pain in a convenience sample of 76 patients who were terminally ill with cancer. Analysis of the McGill Pain Questionnaire Part 1 delineated the following mean scores: total score (11.24), number of words chosen (5.89), sensory (5.71), affective (3.07), evaluative (1.09), and miscellaneous (1.96). The Present Pain Intensity Scale revealed a mean of (1.66) for Part 2. Descriptive statistics were used to analyze choices from the 78 possible word selections that categorize the four dimensions of the McGill Pain Questionnaire: sensory, affective, evaluative, and miscellaneous. Subjects selected 444 words from these four pain components, and their rank order from the highest to the lowest numbers were sensory (227), affective (90), miscellaneous (75), and evaluative (52). Although a mean Present Pain Intensity score of 1.66 demonstrated good pain management, patients with advanced cancer in home hospice care chose words that described four main pain patterns: "tiring and exhausting" (11.5%), "troublesome and annoying" (8.8%), "dull and aching" (7.7%), and "nauseating and sickening" (6.8%). PMID- 11502039 TI - Home caregivers of the person with advanced cancer: an Australian perspective. AB - Australian palliative care services are predominantly community based, with an emphasis on enabling the person to live at home for as long as possible. Home care of the person with advanced cancer receiving palliative care in the community depends largely on the availability of a family caregiver. Family caregivers are required to assess, monitor, and deliver complex therapeutic interventions such as pain and symptom control, including the administration and adjustment of complex medication regimens. This article reports a study of 42 family caregivers providing home care to persons with advanced cancer. This study sought to describe a number of caregiver variables that may influence reactions to caring roles and caregiver well-being. The study found that family caregivers are significantly involved in symptom management, and that they take on almost total responsibility for routine household tasks. In addition, the caregiver role has a negative impact on caregiver health, schedule, anxiety, and energy. However, caregivers find significant meaning in their role and feel relatively well prepared for caregiving. The results of this study support the applicability of the vast international literature on caregiver issues for the Australian setting and suggest the need to move toward development of caregiver-focused nursing interventions. PMID- 11502040 TI - Hope among patients with cancer hospitalized for bone marrow transplantation: a phenomenologic study. AB - This article describes a phenomenologic study designed to further an understanding concerning the concept of hope in a purposive sample of nine patients with cancer hospitalized for bone marrow transplantation. The strategies these patients use to sustain and foster hope are examined. Data collection consisted of a one-time semistructured interview using open-ended questions. A phenomenologic-hermeneutic approach to data analysis was used to identify major themes. The findings showed that participants used six strategies to foster their hope during preparation for BMT: feeling connected with God, affirming relationships, staying positive, anticipating survival, living in the present, and fostering ongoing accomplishment. Religious practices and family members were the most frequently identified sources of hope. The findings of this study provide a base for the improvement of nursing practice. PMID- 11502041 TI - Breast self-exams by teenagers: outcome of a teaching program. AB - Teaching breast self-care as breast changes occur in the adolescent girl can influence positive behaviors such as performing breast self-examinations and seeking regular professional breast examinations. Health promotion behaviors often are taught in high school, but little research has been performed on teaching breast health in a high school setting. Therefore, this descriptive study aimed to identify the beliefs, knowledge, and practices of breast self examination in adolescent girls. A pre- and posttest design was used to determine whether a teaching program would change beliefs, knowledge, and practices of breast self-examination. The results of this study demonstrate that a one-time intervention can be successful in increasing breast self-examination practice and the knowledge of breast self-examination and cancer in adolescents. Before the intervention, approximately 66% of students said they never examined their breasts, and only 3% performed breast self-examination monthly. In 1 month after teaching, these percentages changed appreciably, with 32% of students reporting that they never practiced breast self-examination and 23% reporting that they practiced breast self-examination monthly. PMID- 11502042 TI - Psychometric evaluation of Breast Health Behavior Questionnaire: Spanish version. AB - The purpose of the study was to test the psychometric properties of a culturally sensitive and theory-based instrument: the Breast Health Behavior Questionnaire. This instrument was translated into Spanish and back-translated at a third- to fourth-grade reading level. The pilot group consisted of 70 Hispanic women who attended a class at a local church. Subsequent to pilot testing, another 40 Hispanic women who attended a class at the local health department comprised the study sample. The participants responded to the 15-item questionnaire, which is formatted as a Likert scale. Content validity of the Breast Health Behavior Questionnaire was determined by a panel of experts. A factor analysis of this instrument showed five separate dimensions accounting for 71.82% of the instrument's variance. The three major components of self-regulation theory (schema, coping, and appraisal criteria) were found clustered within the first three dimensions after three items were discarded. The Breast Health Behavior Questionnaire demonstrated an internal consistency reliability coefficient of .7172. The psychometric properties of the Spanish version of this questionnaire warrant further research. The instrument may support a better understanding of the Hispanic woman's practice of breast health behavior. Eventually, the Breast Health Behavior Questionnaire may assist nurses in the formulation of culturally grounded interventions. PMID- 11502043 TI - Attitudes of patients with breast and prostate cancer toward complementary therapies in Finland. AB - This study aimed to describe the attitudes of patients with breast cancer and those with prostate cancer toward complementary therapies. The data were collected with a postal questionnaire administered to 216 patients with breast cancer (response rate, 55.4%) and 1 90 patients with prostate cancer (response rate, 54.9%) in southern and southwestern Finland. The questionnaire was composed of 44 Likert-type statements, which were analyzed using descriptive statistics, chi2 tests, t tests, and two-way analyses of variance. More than half (54%) of the patients with breast cancer and 45% of the patients with prostate cancer believed that people with cancer may benefit from complementary therapies, although they did not think these therapies actually could cure cancer. More than half of the respondents were dubious about using complementary therapies as long as there was no solid scientific evidence. Most believed that complementary therapies were used because they gave people hope (women, 88%; men, 72%) or "something to cling to" (women, 83%; men, 76%). The respondents believed most in dietary therapies and least in healing. There was much confusion and uncertainty about the professional competencies and expertise of the people who provided complementary therapies. Approximately one fourth of the respondents had spoken to their physician about complementary therapies. Only a few had talked about the matter with nursing staff. About half of the respondents thought that physicians and nurses took a negative attitude toward complementary therapies. In both groups, patients who had talked with their physician about complementary therapies tended to show a more positive attitude. PMID- 11502044 TI - Nonfluoroscopic endoventricular electromechanical three-dimensional mapping: current status and future perspectives. PMID- 11502045 TI - Correlation between the effective refractory period and activation-recovery interval calculated from the intracardiac unipolar electrogram of humans with and without dl-sotalol treatment. AB - In experimental studies and/or human body surface mapping, the activation recovery interval (ARI) is used as a parameter to estimate local repolarization. However, it has not been clarified whether the ARI calculated from the intracardiac unipolar electrogram of humans reasonably represents the local effective refractory period (ERP). Measurement of ARIs at multiple ventricular sites can be helpful in assessing the dispersion of ventricular refractoriness of humans, so we examined the relationship between ERP and ARI in the control state and under treatment with dl-sotalol during clinical electrophysiologic studies (EPS). Of 19 patients, an EPS was performed in the control state in 12 and during treatment with dl-sotalol in the other 7. Quadripolar electrode catheters with an interelectrode distance of 5 mm were placed at the right atrium and in the right ventricle. Using atrial pacing, the heart rate was increased incrementally by 10 beats/min, and ERP and ARI were measured for each pacing rate. The ERP at the right ventricle was measured by single extrastimulation between the first and third distal electrodes of the catheter in the right ventricle, and the ARI was calculated from the second distal unipolar electrode of the same catheter as the interval between the minimum derivative of the intrinsic deflection and the maximum derivative of the T wave. In all patients, the unipolar electrogram was stable during the entire EPS, and 83 data points in the control group and 50 in the dl-sotalol group were analyzed. At each pacing rate, the beat-to-beat difference of ARI was less than 10 ms. As the atrial pacing rate increased, the ERP and ARI were progressively shortened, and linear regression analysis revealed an excellent correlation between ERP and ARI. At the same pacing rate, the ERP and ARI in the dl-sotalol group were longer than those in the control group, but no difference was observed in the slope (close to 1.0) and in the intercept of the regression lines between ERP and ARI. In the human ventricle, the ARI calculated from the intracardiac unipolar electrogram represents the local ERP both in the control state and under treatment with dl-sotalol. The ARI can be used as a parameter of local refractoriness and used to study the distribution of refractoriness in the human ventricle. PMID- 11502046 TI - Differences in inflammatory activity at the onset of acute myocardial infarction according to the clinical presentation of preinfarction angina. AB - It is unknown whether the pathogenetic mechanisms underlying acute myocardial infarction (AMI) differ according to the clinical presentation of preinfarction angina, so the present study measured plasma levels of C-reactive protein (CRP) in 280 patients with AMI in whom serum creatine kinase levels were normal on admission and increased subsequently. Patients were classified into 3 groups according to the type of preinfarction angina: no angina (n=95), stable angina (n=48), and unstable angina (n= 137). Patients with unstable angina were subdivided according to the Braunwald classification: class IB (n=39), class IIB (n=22), and class RIB (n=76). There were no differences among the 5 groups in baseline characteristics. CRP on admission was significantly higher and the level of physical activity at symptom onset was significantly lower in the Braunwald class RIB group than in the other groups, but no differences were observed among the other groups. Patients with preinfarction Braunwald class IIB unstable angina had higher CRP levels on admission and symptom onset at a lower level of physical activity. In such patients, the pathogenetic mechanisms may differ from those in other subsets of patients with AMI and active inflammation may play a more important role in AMI onset. PMID- 11502047 TI - Changes in the occurrence of mechanical alternans after long-term beta-blocker therapy in patients with chronic heart failure. AB - Mechanical alternans has been observed in patients with severe congestive heart failure, and the phenomenon is considered to be a terminal sign. Therapeutic strategies for chronic heart failure have significantly developed, but it is uncertain whether patients with mechanical alternans can be effectively treated or not. Seventeen consecutive patients with dilated cardiomyopathy were enrolled: 11 were treated with beta-blockers on conventional therapeutic regimens and 6 patients were not indicated for or were unable to continue beta-blockade. Mechanical alternans was detected during cardiac catheterization in the patients under physiologic tachycardia (110 beats/min) and stepwise dobutamine loading. In the initial study, mechanical alternans occurred in 70.6% of the patients: 8 of the 11 being treated with beta-blockers and 4 of the 6 without beta-blockade therapy. In the second study, none of the patients taking beta-blockers showed mechanical alternans under the same protocol; the occurrence of mechanical alternans did not change in the patients who were not being treated with beta blockers. The left ventricular ejection fraction increased in patients whose mechanical alternans could not be induced during the follow up, but decreased in the patients in whom mechanical alternans was repeatedly inducible. It is concluded that mechanical alternans is associated with the failing myocardium and may be potentially correctable. PMID- 11502048 TI - Prediction of wall motion recovery from the left anterior descending coronary artery velocity pattern recorded by transthoracic doppler echocardiography in patients with anterior wall myocardial infarction retrospective and prospective studies. AB - The diastolic deceleration slope of coronary flow velocity is steeper in patients with substantial 'no reflow' phenomenon than in those without it. This study investigated whether functional outcomes in patients with anterior wall acute myocardial infarction (AMI) can be predicted by analyzing the coronary flow velocity pattern recorded with transthoracic Doppler (TTD) echocardiography. Coronary blood flow velocity in the distal left anterior descending coronary artery was recorded with TTD at day-2 after primary percutaneous transluminal coronary angioplasty/Stent in 51 patients with anterior AMI and the diastolic deceleration half time (DHT, ms) was measured. The wall motion score index (WMSI) was measured at day-1 and -21. In the retrospective study, the DHT was much shorter in those with a poor outcome than in those with good outcome (152 +/- 109 vs 395 +/- 128 ms, p<0.05). Receiver-operating characteristic analysis documented that DHT > or = 300 ms is a suitable cut-off point (sensitivity of 83% and specificity of 93%). In the prospective study (n=30), AWMSI(dl-d21) was significantly higher in those with a DHT > or = 300 ms than those without (0.3 > or = 0.5 vs 1.6 > or = 0.7, p<0.001). DHT correlated significantly with AWMSI(dl d21) (r=0.76, p<0.001). Patients with a shorter DHT of diastolic coronary flow velocity have a poorer functional outcome among patients with anterior AMI. The TTD-determined DHT is a useful predictor of myocardial viability after an anterior AMI. PMID- 11502049 TI - Left ventricular epicardial outflow tract tachycardia: a new distinct subgroup of outflow tract tachycardia. AB - The present study investigated the incidence and ECG characteristics of ventricular tachycardias (VTs) originating from the left ventricular (LV) epicardium. Thirty-one consecutive patients with VT or premature ventricular contraction originating from the outflow tract (OT-VT) underwent catheter ablation. Twenty-one OT-VTs were ablated from the endocardium in the right ventricular (RV) OT and 3 were ablated from the endocardium in the LVOT. In the remaining 7 patients, 4 (13%) OT-VTs were LV epicardial in origin, and 1 of these was ablated from the left sinus of Valsalva. The ECG characteristics of OT-VT of epicardial origin included prominent tall R-waves in the inferior leads, an R wave in V1 and an S-wave in V2, precordial R-wave transition in V2-4, a deep QS wave in aVL, and no S-wave in V6. In addition, there was an atypical left bundle branch block morphology with an inferior axis. These findings were observed during pacing from several sites in the LV epicardium. Furthermore, pacing from the left sinus of Valsalva caused a relatively tall R in V1, deep S-wave in V2 and a tall R-wave with a shallow S-wave in V3, as well as tall R-waves in the inferior leads, which represented intermediate characteristics between RV endocardial OT-VT and LV endocardial OT-VT. In conclusion, OT-VT originating from the LV epicardium is not uncommon and has characteristic ECG findings. Some of them can be ablated from the left sinus of Valsalva. PMID- 11502050 TI - Role of hypertension, dyslipidemia and diabetes mellitus in the development of coronary atherosclerosis in Japan. AB - The present study evaluated the effect of hypertension (HT), dyslipidemia and diabetes mellitus (DM) on the development of coronary atherosclerosis in the Japanese population, using a cross-sectional study of 433 patients (254 men and 179 women) aged 30 years or older who underwent coronary angiography for suspected or known coronary heart disease angina at 5 cardiology departments in the Fukuoka area between September 1996 and August 1997. Patients with a disease duration of 6 months or more were excluded. The main outcome measure was angiographically defined coronary artery stenosis and was found to a significant degree in 146 patients (33.7%). HT, DM, low levels of high-density lipoprotein cholesterol (HDL-C) and hypertriglyceridemia remained as significant coronary artery disease (CAD) risk factors even after controlling for age, sex, hospital, smoking, alcohol use, body mass index and leisure time physical activity. However, hypercholesterolemia was not a significant risk factor after adjusting for these variables. After controlling for these variables, DM, low HDL-C and hypertriglyceridemia were significant CAD risk factors for men, but only DM was a significant CAD risk factor in women. These results indicate that in Japan DM, low HDL-C and hypertriglyceridemia may be more important CAD risk factors than hypercholesterolemia. PMID- 11502051 TI - Differential effects of hypoxia and hypercapnia on respiratory sinus arrhythmia in conscious dogs. AB - To test the hypothesis that hypoxia and hypercapnia have different effects on the genesis of respiratory sinus arrhythmia (RSA), the magnitude of RSA to these stimuli was compared in 3 unanesthetized dogs. Respiration was continuously monitored through a permanent tracheostomy, and the electrocardiogram and blood pressure were also monitored. The magnitude of RSA was assessed as an instantaneous amplitude of the R-R interval oscillation in the high-frequency band of 0.15-0.80Hz by means of complex demodulation. In a total of 47 runs with hypoxia, heart rate, mean arterial pressure, respiratory rate and tidal volume increased, but RSA magnitude decreased even after adjusting for the effects of respiratory rate and tidal volume. In a total of 39 runs with hypercapnia, heart rate and mean arterial pressure did not change, despite the increased respiratory rate and tidal volume. In contrast to hypoxia, RSA magnitude increased even after adjusting for the effects of respiratory rate and tidal volume. The different effects of the two respiratory stimuli on RSA magnitude were noted at any level of ventilation and support the original hypothesis. PMID- 11502052 TI - Apoptosis, rather than oncosis, is the predominant mode of spontaneous death of isolated adult rat cardiac myocytes in culture. AB - A number of isolated adult cardiomyocytes dies within a few days of culture and the mode of death has recently been suggested to be apoptosis, based on its association with the appearance of DNA fragmentation. However, morphological evidence is still lacking and precise analysis, including quantification, has not been performed. Adult rat ventricular cardiomyocytes isolated by enzymatic dissociation were incubated for 7 days in a serum-free medium (the rapid attachment model) and after various incubation periods, both attached and floating cells were counted and classified based on combined criteria of morphology and membrane permeability (dye exclusion): type 1, rod cells with intact membranes; type 2, non-rod cells with intact membranes; and type 3, non rod cells with ruptured membranes. The number of both rod-shaped and dye excluding cells decreased with the incubation period. After 7 days culture, the number of residual cells decreased to 12% of the initial value. Electron microscopy identified type 1 cells as viable, type 2 cells as viable or apoptotic, and type 3 as undergoing oncosis (primary necrosis) or secondary post apoptotic necrosis. Ultrastructural morphometry revealed that oncotic cell death occurred predominantly during the early phase of culture whereas the more abundant apoptotic cell death occurred throughout the culture period. In conclusion, although both apoptotic and oncotic death occur in the natural course of adult rat cardiomyocytes in short-term culture, apoptosis is more predominant. Because of the high incidence of spontaneous cell death predominantly via apoptosis, this information is important for the interpretation of studies using this cell type in culture. PMID- 11502053 TI - Influence of cyclic variation of right ventricular volume on left ventricular mechanical parameters measured with conductance catheter. AB - The conductance catheter is widely used for the continuous measurement of the left ventricular (LV) pressure-volume loops. Cyclical change of the right ventricular (RV) volume may alter the parallel conductance volume, thereby affecting the LV mechanical parameters. Using 8 open-chest adult mongrel dogs, multiple LV pressure-volume loops were obtained by 2 methods: first with a vena cava occlusion (VCO) method, which involved RV volume alteration, and second with a right-heart-bypass (RHB) preparation, which decompressed the right ventricle completely. The slope of the end-systolic pressure-volume relation (Ees), the end systolic volume associated with the end-systolic pressure of 100 mmHg (V100,es), stiffness constant (beta), and the end-diastolic volume associated with the end diastolic pressure of 9 mmHg (V9,ed) were calculated from each loop. There was minimal influence from RV volume alteration on systolic-phase indices [Ees (VCO method, 6.37 +/- 1.91 mmHg/ml; RHB preparation, 6.60 +/- 1.66mmHg/ml; p=0.356), and V100,es (VCO method, 18.4 +/- 9.3ml; RHB preparation, 17.8 +/- 9.0 ml; p=0.681)], but there was a significant influence on diastolic-phase indices [beta (VCO method, 0.0599 +/- 0.0152; RHB preparation, 0.0839 +/- 0.0150; p=0.007), and V9,ed (VCO method, 35.6 +/- 11.3 ml; RHB preparation, 31.9 +/- 12.3 ml; p=0.001)]. The increase in the RV volume in the diastolic phase increased the parallel conductance volume, causing overestimation of the LV diastolic volume measured by the conductance catheter. PMID- 11502054 TI - Left internal mammary-to-left circumflex coronary artery collateral pathway in a patient with occluded left circumflex artery. AB - A unique case of left internal mammary artery-to-circumflex coronary artery collateral pathway, which was conceivably encouraged to develop by coronary occlusion. The pathway provided sufficient collateral perfusion to preserve the local myocardial performance. PMID- 11502055 TI - Recurrence of idiopathic thromboembolism during anticoagulant therapy. AB - Patients with acute pulmonary embolism and venous thromboembolism are usually treated with anticoagulant therapy for at least 3 months as the optimum duration. A patient with recurrent idiopathic venous thromboembolism at the eighth month during anticoagulation (warfarin to target international normalized ratio of 2.0 3.0) is described. The case suggests that patients with idiopathic venous thromboembolism have a high risk of recurrence, even if a strict anticoagulant regimen is followed. PMID- 11502056 TI - Torsade de pointes with a normal QT interval associated with hypokalemia: a case report. AB - The patient was a 46-year-old man with a history of syncope attack after diarrhea. Nonsustained polymorphic ventricular tachycardia (PVT) initiated by short-coupled premature ventricular complex was detected by Holter monitoring. No organic heart disease was found, and the QT interval during sinus rhythm was normal. It was thought that the PVT might be related to hypokalemia, so electrophysiological studies were performed under the condition of hypokalemia (K=3.4mmol/L), after potassium loading (K=4.2mmol/L) and after oral amiodarone therapy. Under the condition of hypokalemia, nonsustained PVT occurred spontaneously, and the monophasic action potential duration at 90% repolarization (MAPD90) at the right ventricular apex was very short (175 ms). The MAPD90 returned to normal after loading potassium (230ms) and after oral amiodarone therapy (240ms), and PVT no longer occurred. With continued oral amiodarone and spironolactone therapy, the patient has been free of syncope attack over a follow up period of 5 years. PMID- 11502057 TI - Right ventricular metastasis from a primary cervical carcinoma. AB - A 49-year-old female presented with a 1-month history of cough and low-grade fever. Echocardiography showed a large mass in the right ventricle and percutaneous right ventricular endomyocardial biopsy provided the histologic diagnosis. Despite radiotherapy and chemotherapy, the patient died. At autopsy, the metastatic deposit in the heart was larger than the primary cervical carcinoma. PMID- 11502058 TI - Host plant effects on activity of the mitosporic fungi Beauveria bassiana and Paecilomyces fumosoroseus against two populations of Bemisia whiteflies (Homoptera: Aleyrodidae). AB - Laboratory bioassays were conducted to determine the effect of host plant on mycosis in two geographically distinct populations of early 2nd-instar nymphs of Bemisia argentifolii Bellows & Perring from the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin and Paecilomyces fumosoroseus (Wize) Brown & Smith. Mycosis in B. argentifolii nymphs varied according to the host plant on which the nymphs were reared but not according to the population. Both populations of whiteflies reared on cotton were consistently significantly less susceptible to infection by either fungus than when reared on melon. We hypothesized that the cotton plant produced a fungal inhibitor that may confer protection on whiteflies feeding (and possibly sequestering) upon it. Germination of conidia of both fungi was strongly inhibited (below 12% germination) on the cuticle of nymphs reared on cotton but was over 95% on the cuticle of nymphs reared on melon. We further hypothesized that the terpenoid gossypol, produced by many cultivars of cotton, might have been involved in antibiosis. Gossypol mixed with Noble agar at five concentrations was tested for its effects on germination of conidia of both fungi. P. fumosoroseus was highly tolerant of gossypol, even at the relatively high concentration of 1000 ppm, while B. bassiana tolerated gossypol at concentrations up to 500 ppm and strong inhibition only occurred in presence of gossypol at 1000 ppm. Our in vivo findings on cotton and on the insect's cuticle pointed at a potential host plant-mediated antibiosis. The in vitro tolerance of P. fumosoroseus and partial tolerance of B. bassiana to gossypol disagreed with our in vivo data. Gossypol concentrations higher than 1000 ppm might have increased the sensitivity of the fungi in our in vitro tests. Sequestered gossypol (and/or other cotton plant allelochemicals) by B. argentifolii nymphs would explain, at least partially, the insect's defense against the pathogens. PMID- 11502059 TI - Effects of prolonged oral administration of fumonisin B1 and aflatoxin B1 in rats. AB - The effects of prolonged oral administration (21 days) of fumonisin B1 (FB1) and aflatoxin B1 (AFB1) were evaluated on male Wistar rats. The animals were housed in individual metabolic cages and submitted to the following treatments: 1-0 microg AFB1 + 0 mg FB1/100g bw.; 2-72 microg AFB1+ 0 mg FB1/100 g bw; 3-0 microg AFB1 + 0.5 mg FB1 g bw; 4-0 microg AFB1 + 1.5 mg FB1/100 g bw; 5-72 microg AFB1 + 0.5 mg FB1/100g bw; 6-72 microgAFB1 + 1.5 mg FB1/100g bw. On day 21, the rats were sacrificed for evaluation. The results showed that treated animals presented differences in body weight and absolute/relative weights of liver and kidney as well as altered hepatic function and cholesterol blood levels. Rats fed with the greatest doses of AFB1 and FB1 gained less weight (2.79 g/day) at the end of the experimental period; their blood concentrations of liver enzymes aspartate aminotransferase (AST) and alkaline phosphatase (AP) were above control levels (130.35 micro/l and 471.00 micro/l, respectively). Blood cholesterol increased in the groups treated with the highest dose of FB1 or FB1 associated with AFB1. Histopathology revealed the occurrence of apoptosis in the liver of rats exposed to FB1. The association of aflatoxin B1 with fumonisin B1 at higher dose probably potentiated the effects of the higher dose of fumonisin B1 acting singly. PMID- 11502060 TI - Isolation of aflatoxigenic strains of Aspergillus and detection of aflatoxin B1 from feeds in India. AB - In a preliminary study, 256 feed samples collected from different parts of Northern India were examined for the presence of aflatoxigenic strains of Aspergillus favus/parasiticus and for detection of Aflatoxin B1 (AFB1). Out of 198 A. flavus and 15 A. parasiticus strains isolated, 76% and 86% respectively, were found to be toxigenic. Aflatoxin B1 content of these feeds, as estimated by thin layer chromatography (TLC) and enzyme linked immunosorbent assay (ELISA) were very high (average 0.412+/-0.154 ppm) in comparison to the permissible Indian regulation level (0.03 ppm). Seasonal variation of incidence and level of toxin in feed was recorded and it was high during monsoon/post monsoon period. PMID- 11502061 TI - Microcystis aeruginosa as the causative organism of rhinosporidiosis. PMID- 11502062 TI - Cost-effective multiplication of the entomopathogenic fungus Nomuraea rileyi (F) Samson. AB - Cost-effective and rapid multiplication of Nomuraea rileyi is reported. The spore yields in semi-synthetic media were comparable or significantly higher to the standard medium. Maltose and peptone, carbon and nitrogen sources could be effectively replaced with 2% barley extract and 1% soybean extract respectively. However, replacement of yeast extract with dry yeast resulted in lower spore yields. Sporulation of the fungus multiplied on solid substrate was possible only when the bags used had a 0.2 microm filter to facilitate passive exchange of sterile air. A high spore yield of 2.8 x 10(9)/g of substrate was realized on crushed sorghum. PMID- 11502063 TI - Fungal flora of the digestive tract of 5 species of triatomines vectors of Trypanosoma cruzi, Chagas 1909. AB - A study of the mycobiota in the digestive tract of 5 important species of triatomines, Triatoma brasiliensis, T infestans, T. sordida, T. pseudomaculata and T. vitticeps, was made. The digestive tracts of 164 adults and 535 nymphs of those triatomines were studied and 393 fungal strains were isolated. The genera with the greatest number of species were Penicillium (19 species), Aspergillus (17 species) and Acremonium (5 species) and the most frequent species, in decreasing order, were Penicillium corylophilum, Aspergillus niger, Penicillium felluttanum, Cladosporium herbarum, Penicillium waksmanii, Aspergillus awamori and Paecilomyces variotii. Among the isolated fungi, we found species that are recognized as entomopathogenic and pathogenic for humans and animals. PMID- 11502064 TI - Susceptibility profile of vaginal yeast isolates from Brazil. AB - Vaginal specimens for culture were obtained from two hundred and five immunocompetent, non-hospitalized patients selected among all women attending the Gynecology and Obstetric Ambulatory Clinic of the University of Espirito Santo, Brazil, during a 2-year period (From 1998 to 1999). Patients were checked for signs and symptoms of vulvovaginitis and previous use of topical and systemic antifungal drugs. Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to NCCLS microbroth assay. The prevalence of vaginal yeast isolates from asymptomatic women was 25% (30/121) and 60% (50/84) among patients with symptoms of vulvovaginitis. Candida albicans was the most frequently isolated species in both groups (46% and 90%, respectively), followed by C. glabrata (13% and 6%, respectively). All isolates were susceptible to amphotericin B. Only ten isolates had dose dependent susceptibility (DDS) or resistance to azoles; and seven of these were non albicans species. Based on our results we suggest that species identification and antifungal susceptibility testing need not be routinely performed in immunocompetent women, and may be reasonable only for the minority of patients with complicated vulvovaginal candidiasis that fail to respond to therapy. PMID- 11502065 TI - Cytokines and chemokines. PMID- 11502066 TI - Receptor dimerization: a key step in chemokine signaling. AB - Chemokines exert their effects through their interaction with seven transmembrane domain receptors coupled to G-proteins, GPCRs. Such receptor ligation leads to the regulation of numerous activities where chemokines play a key role, including hematopoiesis, T-cell activation, angiogenesis, inflammatory diseases or HIV-1 infection. Here we discuss the molecular mechanisms that underlie chemokine receptor activation. As occurs with other GPCRs, chemokines initiate the signaling cascades by inducing receptor dimerization. This dimerization enables the activation of the JAK/STAT pathway which allows the subsequent triggering of G-protein dependent signaling events. This mechanism provides a new context to explain some of the activities exerted by chemokines and introduces new targets for the development of drugs to fight those diseases were chemokines are implicated, such as inflammation and AIDS. PMID- 11502067 TI - The soluble IL-6 receptors: serum levels and biological function. AB - IL-6 exerts its biological activities through interaction with specific receptors expressed on the surface of target cells. IL-6 binds first to a low-affinity (10( 9) M) subunit, a 80 kDa glycoprotein also called gp80 or IL-6R alpha. The IL-6/IL 6R alpha complex recruits the signal-transducing b subunit, a 130 kDa glycoprotein called gp130. The association of gp130 with IL-6 and IL-6R alpha leads to the formation of the high-affinity IL-6 receptor complex, to the linkage of two gp130 subunits and to signal transduction. Soluble forms of both receptors have been described and found in biological fluids. Soluble cytokine receptors are generated by either proteolytic cleavage of their membrane moiety or by alternative splicing. Both mechanisms have been described for sIL-6R and sgp130 formation. Interestingly, the association of IL-6 with the soluble form of IL-6R alpha is capable of eliciting a biological response in cells that express only the membrane gp130. This type of activation, called "trans-signalling", renders virtually all cells capable of responding to IL-6/sIL-6R alpha complexes, making for a large new spectrum of IL-6 activities, ranging from the control of the immune response to involvement in pathological states. In this review the biological activities of IL-6 will be considered in the light of new knowledge concerning the association of IL-6 and the soluble IL-6 receptors. PMID- 11502068 TI - Chemokines and lymphocytes: the role of chemokines and their receptors in the immune system. AB - In the last few years. chemokines have emerged as an important superfamily where importance extends far beyond their most famous function as inflammatory mediators. Indeed, they are important molecules not only in inflammatory responses but also as immunoregulators. Chemokines ensure the continuous recirculation of immune cells among the various anatomical microenvironments, and are essential for maintaining immunological homeostasis. In addition, chemokines also have critical functions in lymphocyte development. In this article, we review the role of chemokines and their receptors in lymphopoiesis, lymphocyte's migration and immune response. PMID- 11502069 TI - Interferon-alpha in inflammation and immunity. AB - Type I interferons, constituting IFN-alpha and IFN-beta, were initially described for their ability to interfere with viral replication. IFN-alpha was the first cytokine to be cloned and used successfully as a therapeutic cytokine, although its mechanism of action remained largely elusive. Evidence gathered over the last few years shed light on the molecular effects of IFN-alpha, especially its interaction with the cytokine cascade. Recently, the principle source of IFN alpha could be identified as the precursor of type 2 dendritic cells, and IFN alpha has been identified as the cytokine linking innate with adaptive immunity via its interaction with dendritic cells. PMID- 11502070 TI - TNF and TNFR biology in health and disease. AB - Many insights have been gained into cytokine-regulated control of inflammatory processes and host defence in recent years. Evidence has also gradually accumulated that cytokine cascades play a central role in events regulating cell death and differentiation. Further developments include an understanding that the biological effects of the tumor necrosis factor-alpha (TNF-alpha or TNFSF) cytokine may be regulated by soluble TNF receptor binding and that modulation of receptor levels may permit physiological inhibition of TNF action. There has been a gradual realisation of the value of TNF/TNFR ratios as predictors of disease outcome, and the discovery of functional regulatory polymorphisms of the TNF gene and mutations of TNFRSF1A (TNFR1 receptor) have led to conceptual breakthroughs in our understanding of the genetic control of inflammation. However the exact mechanisms by which TNFRSF1A mutations give rise to disease susceptibility are not yet well understood. Over the past 10 years these concepts have been used as the basis for successful anti-TNF therapy of autoimmune diseases like rheumatoid arthritis (RA) and Crohn's disease. PMID- 11502071 TI - Role of interleukin-13 in innate and adaptive immunity. AB - Initially thought to be functionally redundant with IL-4 as a predominant anti inflammatory factor secreted during type-2 T-cell responses, IL-13 possesses a number of additional properties that distinguish it from IL-4 in addition to having both anti-inflammatory and immune activating properties. This review centers primarily on the role of IL-13 in the regulation of cellular functions of innate immunity and acquired immunity against certain microbial pathogens. First, we discuss IL-13's regulation of innate cell targets and its impact on inflammation, antigen uptake and antigen presentation. Second, we focus on IL 13's involvement in acquired immunity to infectious helminths and protozoa. The role of this cytokine in immune responses is still being determined but evidence to date suggests this molecule has been conserved as an important regulatory factor involved in both early innate and late adaptive responses. PMID- 11502072 TI - The impact of chemokine receptor conformational heterogeneity on HIV infection. AB - Receptor binding largely governs viral tropism, since the presence of CD4 and an appropriate coreceptor is a prerequisite for membrane fusion and virus infection. Env-receptor interactions are conformationally complex, involving multiple regions in both gp120 as well as in the receptors. As a result, differences in receptor conformation, posttranslational processing, and surface density all have the potential to influence viral infectivity and therefore tropism and pathogenesis. This review gives an overview of the research that led to the discovery of chemokine receptors as coreceptors for HIV-1, describes the repertoire of coreceptors described to date and addresses their in vivo relevance. We will discuss very recent studies that indicate that while the presence of CD4 and coreceptor are necessary for virus infection, their mere presence is not sufficient. PMID- 11502073 TI - Mediators of inflammation and acute phase response in the liver. AB - The acute phase response is a generalized response of the organism to multiple disturbances of its physiological homeostasis. It consists of local and systemic reactions. Inflammatory processes are the main causes for the initiation of these defence mechanisms. Responsible mediators for the acute phase response are predominantly cytokines, whereby the liver is the predominant target organ. Changes in hepatocyte gene expression profiles result in dramatic changes in serum concentrations of specific plasma proteins, called acute phase proteins. IL 6 was identified as the principal mediator of this reaction. Via its cellular signal transducer gp130 IL-6 induces DNA-binding of STAT transcription factors on regulatory elements of target genes. While IL-6 dependent processes are mainly conferred to be protective other inflammatory cytokines are attributed to be cytotoxic for the liver. TNF-alpha was shown to be involved in several models of liver failure as a mediator for both cytotoxicity and cell proliferation. TNF alpha leads via caspases to the onset of apoptosis, the so-called programmed cell death. On the other hand it activates NF-kappaB thereby triggering inflammatory processes. In this review we display the relevance for intracellular actions of both cytokines in several models of liver injury. Especially we refer to the T cell mediated Concanavalin A induced liver failure and to liver regeneration induced by CCL4 and partial hepatectomy. Both cytokines contribute in concert to a cellular balance during these pathophysiological conditions. PMID- 11502074 TI - Cytokines in rheumatoid arthritis: is it all TNF-alpha? AB - Anti-TNF-alpha therapy has shown clear efficacy in the treatment of rheumatoid arthritis (RA). Since some patients do not respond and the treatment is suspensive, combination therapy may be of interest. Other cytokines produced by monocytes such as IL-1, IL-12, IL-18 are also involved. The secretion of these cytokines is regulated by subsets of T-lymphocytes. Among these, IL-17 producing Th1 cells appear to contribute directly to the destructive process. Furthermore, this T-cell contribution enhances the action of monocyte derived proinflammatory cytokines. Using models of human RA synovium inflammation and bone resorption, ex vivo results suggest that combination therapy may be of interest. Acting on more than one cytokine may increase the percentage of responding RA patients as well as the degree of individual patient response. PMID- 11502075 TI - Cytokines, from atopy to asthma: the Th2 dogma revisited. AB - Asthma is a spreading condition in Western countries, in most cases in relationship with atopy. Atopy is defined by an individual predisposition to develop allergic diseases in response to environmental allergens. The atopic immune system is characterized by a Th2 deviation determined by genetic and environmental factors. Among these factors, the role of allergen exposure, dietary behavior, air pollution and early exposure to microbes is discussed. In asthma, a Th2 cell activation is evident, but is accompanied by a Tc1 cell activation. These Tc1 cells probably down-regulate Th2 cells, but are also relevant to the bronchial hyperresponsiveness characterizing asthma. We propose that Tc1 activation in asthma could be the link between allergy and bronchial hyperresponsiveness. PMID- 11502076 TI - Breach of IL-12 monopoly in the initiation of type 1 immunity to intracellular infections: IL-12 is not required. AB - IL-12 is believed to play an important role in type 1 T-cell differentiation and type 1 cytokine IFN-gamma release by T- and NK-cells and macrophages in host defense against intracellular infections by bacteria, parasites, fungi and viruses. However, recent studies by us and others have provided unequivocal evidence that while IL- 12 is critically required for the development of type 1 immunity to the majority of intracellular bacterial, parasitic and fungal infections, it is not required for anti-viral type 1 immune responses. These findings have provoked our re-thinking about the role of IL-12 in type 1 immunity and the search for additional cytokines capable of initiating anti-viral type 1 immunity. We hypothesize that there exist multiple cytokines including IL-12 which play a redundant role in the initiation of type 1 immunity against viral infection. These cytokines are likely released from not only antigen-presenting macrophages/dendritic cells but many other cell types, which suits the mode of viral infection. The existence of multiple factors capable of driving type 1 immunity endows the host with additional safeguards to cope with prevalent viral foes. PMID- 11502077 TI - Pro- versus anti-inflammatory cytokines: myth or reality. AB - Inflammation is characterized by an interplay between pro- and anti-inflammatory cytokines. Cytokines are commonly classified in one or the other category: interleukin-1 (IL-1), tumor necrosis factor (TNF), gamma-interferon (IFN-gamma), IL-12, IL-18 and granulocyte-macrophage colony stimulating factor are well characterized as pro-inflammatory cytokines whereas IL4, IL-10, IL-13, IFN-alpha and transforming growth factor-beta are recognized as anti-inflammatory cytokines. In this review, we point out that this classification is far too simplistic and we provide numerous examples illustrating that a given cytokine may behave as a pro- as well as an anti-inflammatory cytokine. Indeed, the cytokine amount, the nature of the target cell, the nature of the activating signal, the nature of produced cytokines, the timing, the sequence of cytokine action and even the experimental model are parameters which greatly influence cytokine properties. PMID- 11502078 TI - Characterization of an IL-2 mimetic with therapeutic potential. AB - Human interleukin-2 (IL-2) interacts with two types of functional receptors (IL 2R alpha betagamma and IL-2R betagamma) and acts on a broad range of target cells involved in inflammatory reactions and immune responses. IL-2 is also used in different clinical trials aimed at improving the treatment of some cancers and the recovery of CD4 lymphocytes by HIV patients. The therapeutic index of IL-2 is limited by various side effects dominated by the vascular leak syndrome. We have shown that a chemically synthesised fragment of the IL-2 sequence can fold into a helical tetramer likely mimicking the quatemary structure of an hemopoietin. Indeed, peptide p1-30 (containing amino acids 1 to 30, including the sequence corresponding to the entire alpha helix A of IL-2) spontaneously folds into an alpha-helical homotetramer and stimulates the growth of T-cell lines expressing human IL-2R beta, whereas shorter versions of the peptide lack helical structure and are inactive. At the cellular level, p1-30 induces lymphokine-activated killer (LAK) cells and preferentially activates CD8 low lymphocytes and natural killer cells, which constitutively express IL-2R beta. A significant IFN-gamma production is also detected following p1-30 stimulation. A mutant form of p1-30 (Asp20-->Lys) which is likely unable to induce vascular leak syndrome remains capable to generate LAK cells like the original p1-30 peptide. Altogether our data suggest that p1-30 has therapeutic potential. PMID- 11502079 TI - Diagnostic differences in social anhedonia: a longitudinal study of schizophrenia and major depressive disorder. AB - This study examined the hypothesis that, in schizophrenia, elevated trait social anhedonia (SA) is a stable individual difference, whereas in depression, increased SA is a reflection of a current clinical state that will diminish with recovery. Differences in trait Negative Affect (NA) and Positive Affect (PA) were also examined. Individuals with schizophrenia (n = 55) and depression (n = 34) were evaluated at baseline during hospitalization and compared with nonpsychiatric control participants (n = 41). Participants were assessed again at a 1-year follow-up. At baseline, compared with control participants, individuals with schizophrenia and depression were both characterized by elevated SA, greater NA, and lower PA. In schizophrenic individuals, elevated SA remained stable over the follow-up. However, in recovered depressed patients, SA declined over the follow-up period. Group differences remained in NA and PA over the 1-year follow up. These results support the view that elevated SA is enduring in schizophrenia but that elevated SA is transiently related to clinical status in depression. PMID- 11502080 TI - Anxiety sensitivity, panic, and depressed mood: a reanalysis teasing apart the contributions of the two levels in the hierarchial structure of the Anxiety Sensitivity Index. AB - Anxiety sensitivity (AS) has been defined as the fear of anxiety and anxiety related sensations, and evidence suggests that AS plays an important role in the psychopathology of panic. It is entirely unclear whether the relation between AS and panic should be attributed to one (or more) of the AS group factors, the general AS factor, or to factors at both levels of the AS hierarchy. The authors reanalyzed data presented earlier by R. M. Rapee, T. A. Brown, M. A. Antony, and D. H. Barlow (1992) to tease apart the contributions of the different levels of the AS hierarchy to fear responses to hyperventilation and 5.5% carbon dioxide challenges. The results demonstrated that AS-Physical Concerns is the only one of the three AS group factors that contributes to relations with fear responses to these two challenges. However, AS-Mental Incapacitation Concerns had a stronger positive linear association with depressed mood than did AS-Physical Concerns. PMID- 11502081 TI - Transitioning into and out of large-effect drinking in young adulthood. AB - As individuals age beyond the college years into young adulthood, many exhibit a tendency to moderate or "mature out of" alcohol involvement. The current study classified effect-drinking statuses in young adults and examined transitions among statuses using latent transition analysis, a latent variable state sequential model for longitudinal data. At 3 occasions over 7 years (Years 1, 4, and 7), 443 men (47%) and women (mean age of both at baseline = 18.5 years; 51% with family history of alcoholism) responded to 3 past-30-day items assessing drinking and subjective effects of drinking: whether the respondent drank alcohol, felt high, and felt drunk. Latent statuses included abstainers (14% at Year 1), limited-effect drinkers (8%), moderate-effect drinkers (23%), and large effect drinkers (54%). Respondents with family history of alcoholism were less likely to transition out of large-effect drinking than those without family history. Men exhibited more severe initial effect-drinking statuses and lower transition probabilities into less severe effect-drinking statuses than women. PMID- 11502082 TI - Patterns of recovery from trauma: the use of intraindividual analysis. AB - Patterns of recovery from sexual and nonsexual assault were examined. Two studies containing data from female victims of these assaults were analyzed. In Study 1, victims (N = 101) underwent 12 weekly assessments with measures of posttraumatic stress disorder (PTSD), depression, and state anxiety. In Study 2, victims (N = 108) underwent monthly assessments on the same measures. The authors examined the effects of type of trauma and time of peak reaction on long-term recovery using intraindividual analysis of change. In both studies, initial and peak reactions of rape victims were more severe than were those of nonsexual assault victims on all measures of psychopathology. Victims with delayed peak reaction exhibited more severe pathology at the final assessment than did victims with early peak reaction. Results of Study 2 indicated a slower recovery rate from sexual than nonsexual assault; in Study 1 a similar pattern of recovery emerged. The advantages of an individual-focused, longitudinal approach to recovery from a trauma are discussed. PMID- 11502083 TI - Using the five-factor model to represent the DSM-IV personality disorders: an expert consensus approach. AB - This study sought to extend previous work on the five-factor dimensional model (FFM) of personality disorder (PD) by developing more comprehensive FFM descriptions of prototypic cases. Specifically, the authors asked experts in each of the 10 DSM-IV PDs to rate the prototypic case by using all 30 facets of the FFM. Aggregating across raters of the given disorder generated a prototype for each disorder. In general, there was good agreement among experts and with previous theoretical and empirical FFM translations of DSM diagnostic criteria. Furthermore, the ability of the FFM explanation to reproduce the high comorbidity rates among PDs was demonstrated. The authors concluded that, with the possible exception of schizotypal PD, the DSM PDs can be understood from the dimensional perspective of the FFM. Future directions for research, including the use of the present prototypes to "diagnose" personality disorder, are discussed. PMID- 11502084 TI - A taxometric investigation of the latent structure of worry. AB - Researchers have described 2 types of worriers, normal and pathological, who differ in the frequency, intensity, and controllability of their worry experiences. Although normal and pathological worry are generally treated as separate though related phenomena, no study has tested for separateness against the alternative hypothesis that all worry exists along a single dimension. In the present study, worry ratings of 1,588 college students were submitted to taxometric procedures designed to evaluate latent structure. Results provided evidence for the dimensionality of worry. These findings suggest that generalized anxiety disorder (GAD), whose central feature is worry, may also be quantitatively rather than qualitatively different from normal functioning. The authors argue that a focus on normal and pathological extremes has constrained the study of worry phenomena and that dimensional conceptualization of worry may significantly enhance understanding of both worry and GAD. PMID- 11502085 TI - Autonomic stress reactivity and executive functions in successful and unsuccessful criminal psychopaths from the community. AB - A significant gap in the psychopathy literature is the lack of studies comparing "successful," nonconvicted psychopaths with "unsuccessful," convicted psychopaths. This study tested the hypothesis that successful psychopaths show increased autonomic stress reactivity and better neuropsychological function compared with unsuccessful psychopaths. A total of 26 controls, 16 unsuccessful psychopaths, and 13 successful psychopaths were assessed on psychophysiological measures recorded during an emotional manipulation, the Wisconsin Card Sorting Test (WCST), Wechsler Memory Scale--Revised subtests, and childhood stressors. Compared with controls, unsuccessful psychopaths showed reduced cardiovascular stress reactivity. In contrast, successful psychopaths showed heightened reactivity, better WCST performance, and more parental absence than unsuccessful psychopaths and controls. The implications of these findings and the generalizability of existing psychopathy research are discussed. PMID- 11502086 TI - Somatosensory processing in the biological relatives of schizophrenia patients: a signal detection analysis of two-point discrimination. AB - This study examined 2-point discrimination performance in 1 st-degree biological relatives (n = 39) of individuals with schizophrenia and normal adult control participants (n = 30) recruited from the community. They completed an objective 2 point discrimination task, adapted for use with a signal detection approach to permit separation of discriminability (i.e., sensitivity, d') from response bias/ criterion (or motivation, lnbeta). Relatives revealed poorer performance on the d' index compared with controls. The 2 groups did not differ on lnbeta, suggesting a genuine difference in sensitivity but not response bias. The sensitivity deficit might reflect decreased spatial acuity and/or impaired intensity cue processing of tactile stimuli. Poor performance on the d' index was most closely associated with 2 schizotypic features, namely "odd beliefs/magical thinking." PMID- 11502087 TI - Long-term stability of communication deviance. AB - Communication deviance (CD), forms of communication that are not bizarrely thought disordered but are hard to follow and that make difficult the consensual sharing of attention and meaning, has been hypothesized as a nonspecific contributor of rearing parents to psychopathology of offspring, including schizophrenia. This hypothesis, or an alternative of genetic transmission, would gain plausibility if CD has long-term stability. CD was evaluated, using tape recorded and reliably scored Rorschachs in 158 Finnish adoptees, and retested after a median interval of 11 years. Adolescent CD was not stably correlated with follow-up CD. However, initial CD at a mean age of 32 and follow-up CD were significantly correlated. Gender, genetic risk for schizophrenia, and DSM-III-R (American Psychiatric Association, 1987) psychiatric diagnoses had no effect on adult CD stability. CD appears to be a stable, traitlike feature of adult but not adolescent functioning. PMID- 11502088 TI - Specifying the relations between affect and heavy alcohol use among young adults. AB - Current reformulations of the tension reduction hypothesis posit that only a subset of vulnerable individuals are at risk for drinking in response to negative affect. To further specify this model, this study examined the types of mood and social contexts under which affect and alcohol use are associated. Participants were 74 college students who completed repeated assessments of mood, alcohol use, friendship quality, and social support. A complex pattern of findings supported the moderating influences of gender, friendship factors, and the timing of behavior (i.e., weekends vs. weekdays) on the relation between affect and alcohol use. Young adults with less intimate and supportive friendships, as compared with their peers, showed risk for greater drinking following relative elevations in sadness and hostility. Such drinking episodes, in turn, predicted subsequent elevations in these same negative moods the following week. Gender differences in such a cyclical pattern of affect and alcohol use were found to vary across differing emotional experiences. Recommendations for a more refined theory of affect and alcohol use are discussed. PMID- 11502089 TI - Psychopathy, antisocial personality, and suicide risk. AB - H. Cleckley (1976) maintained that psychopaths are relatively immune to suicide, but substantial evidence exists for a relationship between antisocial deviance and suicidal acts. This study was the first to explicitly examine suicidal history among psychopathic individuals as defined by R. D. Hare's (1991) Psychopathy Checklist--Revised (PCL-R). Male prison inmates (N = 313) were assessed using the PCL-R and DSM-III-R and DSM-IV criteria (American Psychiatric Association, 1987, 1994) for antisocial personality disorder (APD), and they completed A. Tellegen's (1982) Multidimensional Personality Questionnaire (MPQ). Presence or absence of prior suicide attempts was coded from structured interview and prison file records. Suicide history was significantly related to PCL-R Factor 2 (which reflects chronic antisocial deviance) and to APD diagnosis but was unrelated to PCL-R Factor 1, which encompasses affective and interpersonal features of psychopathy. Higher order MPQ dimensions of Negative Emotionality and low Constraint were found to account for the relationship between history of suicidal attempts and antisocial deviance, indicating that temperament traits may represent a common vulnerability for both. PMID- 11502090 TI - Structural relations between borderline personality disorder features and putative etiological correlates. AB - This study assessed the structural relations between borderline personality disorder (BPD) features and purported etiological correlates. Approximately 5,000 18-year-old nonclinical young adults were screened for BPD features, and 2 cohorts of participants (total N = 421; approximately one half of whom endorsed significant borderline features) completed the laboratory phase of the study. Measures included self-report and interview-based assessments of BPD psychopathology, personality, psychopathology in biological parents, and childhood physical and sexual abuse. Significant relations between BPD features and purported etiological correlates of BPD were found. A multivariate model that included parental psychopathology, childhood abuse, and personality factors provided an adequate fit to the data and supported the contention that the personality traits disinhibition and negative affectivity underlie BPD features. PMID- 11502091 TI - Pathological gamblers, with and without substance use disorders, discount delayed rewards at high rates. AB - Pathological gambling is classified as a disorder of impulse control, yet little research has evaluated behavioral indices of impulsivity in gamblers. The rates at which rewards delayed in time are subjectively devalued may be a behavioral marker of impulsivity. This study evaluated delay discounting in 60 pathological gamblers and 26 control participants. Gamblers were divided into those with (n = 21) and without (n = 39) substance use disorders. A hypothetical $1,000 reward was delayed at intervals ranging from 6 hr to 25 years, and immediate rewards varied from $1 to $999. Pathological gamblers discounted delayed rewards at higher rates than control participants, and gamblers with substance use disorders discounted delayed rewards at higher rates than non-substance-abusing gamblers. These data provide further evidence that rapid discounting of delayed rewards may be a feature central to impulse control and addictive disorders, including pathological gambling. PMID- 11502092 TI - Diminished response to pleasant stimuli by depressed women. AB - This study examined the self-report and facial expressions of emotional response to pictorial stimuli and the incidental learning of pleasant and unpleasant words by depressed (n = 20) and nondepressed (n = 20) women. Depression was associated with reports of diminished emotional response and reduced frequency and intensity of facial expressions only to pleasant stimuli. The 2 groups did not differ in response to hedonically unpleasant stimuli, even those specifically relevant to the emotion of sadness. In a similar vein, depressed and nondepressed participants showed differences in incidental recall for only pleasant self referential terms. There was no difference in recall of unpleasant words. These findings suggest the importance of hedonic deficits on psychological processes in clinical depression. PMID- 11502093 TI - Mouse hepatitis virus type-2 infection in mice: an experimental model system of acute meningitis and hepatitis. AB - Infection with mouse hepatitis virus (MHV) strain A59 produces acute hepatitis, encephalitis, and chronic demyelination in mice. However, little is known about a closely related strain, MHV-2, which is only weakly neurotropic. To better understand the molecular basis of neurotropism of MHVs, we compared the pathogenesis and genomic sequence of MHV-2 with that of MHV-A59. Intracerebral injection of MHV-2 into 4-week-old C57B1/6 mice produces acute meningitis and hepatitis without encephalitis or chronic inflammatory demyelination. Sequence comparison between MHV-2 and MHV-A59 reveals 94-98% sequence identity of the replicase gene, 83-95% sequence identity of genes 2a, 3, 5b, 6, and 7, and marked difference in the sequence of genes, 2b, 4, and 5a. This information provides the basis for further studies exploring the mechanism of viral neurotropism and virus induced demyelination. PMID- 11502094 TI - TNF-alpha, PDGF, and TGF-beta(1) expression by primary mouse bronchiolar-alveolar epithelial and mesenchymal cells: tnf-alpha induces TGF-beta(1). AB - The bronchiolar-alveolar epithelium (BAE) is a primary target site for inhaled agents that cause lung injury. These cells, consequently, release a broad range of mediators that influence other cell populations, including interstitial lung fibroblasts that are central to the development of interstitial pulmonary fibrosis (IPF). A number of peptide growth factors (GF) have been postulated to be essential in the pathogenesis of IPF. We demonstrate here that primary populations of mouse BAE and mesenchymal cells, maintained in culture, synthesize four potent GF. These are platelet-derived growth factor isoforms (PDGF) A and B, transforming growth factor beta-1 (TGF-beta(1)), and tumor necrosis factor alpha (TNF-alpha). A mouse lung epithelial cell isolation technique pioneered in this laboratory has been used to purify the BAE cells to greater than 85% (80 +/- 5.6% alveolar type II and 9 +/- 2.3% Clara cells) in culture. Northern analysis, RNase protection assay, and immunocytochemistry (ICC) were used to establish mRNA and protein expression of the GF over time in the cultured BAE and mesenchymal cells. We show for the first time in these primary mouse lung cells that treatment of both cell types with TNF-alpha upregulates expression of TGF-beta(1). The four GF are produced by both epithelial and mesenchymal cells but with different temporal patterns. TGF-beta(1) is expressed constitutively by BAE and mesenchymal cells, whereas TNF-alpha expression wanes over time. The findings by ICC were consistent with levels of mRNA expression in both cell types. As genetically defined and altered mouse strains are becoming increasingly valuable for modeling lung disease, studying the gene expression patterns of target cells from these animals in vitro would be useful in sorting out the complex responses by individual cell types of the lung and the interactions among the multitude of mediators that are released during lung cell injury. PMID- 11502095 TI - cAMP enhances Cx43 gap junction formation and function and reverses choline deficiency apoptosis. AB - Previously, it had been shown that acute choline deficiency (CD) induced apoptosis in cultured rat liver epithelial cells, whereas cells that are adapted to survive in low-choline-containing medium acquire resistance to CD apoptosis and undergo malignant transformation. Thus, understanding the mechanisms of action of CD could increase our understanding of the role of choline, an essential nutrient, in the process of malignant transformation. The present experiments were designed to test the hypothesis that CD might function as a pro apoptotic trigger by altering the localization of connexin 43 gap junction protein and gap junctional intercellular communication (GJIC). Established liver epithelial cells (WB cells; Hep3B cells) were maintained in a defined, serum-free medium control (70 microM choline) or choline deficient medium (CD, 5 microM choline) and the localization of connexin 43 protein (Cx43) was studied by immunocytochemistry and Western blotting. In nontumorigenic WB cells, CD apoptosis was associated with retention of Cx43 in the golgi/ER region of the cytoplasm and decreased GJIC as measured using a preloading fluorescent dye transfer method (calcein AM/DiIC(18)). Cells maintained in CD in the presence of 8-bromoadenosine 3':5'-cyclic monophosphate exhibited restoration of Cx43 at the plasma membrane and increased GJIC and inhibition of apoptosis. These studies show that CD apoptosis in nontumorigenic liver epithelial cells is associated with alterations to Cx43 and GJIC and that an uncoupling of Cx43 localization and GJIC is related to resistance to CD apoptosis in transformed liver epithelial cells. PMID- 11502096 TI - Atherosclerotic lesions are associated with increased immunoreactivity for inducible nitric oxide synthase and endothelin-1 in thoracic aortic intimal cells of hyperlipidemic Watanabe rabbits. AB - The development and progression of atherosclerotic lesions in Watanabe heritable hyperlipidemic rabbits is associated with increases in inducible nitric oxide synthase (NOS2) and endothelin-1 (ET-1) immunoreactivity. In contrast, there is a reduction of immunoreactivity for neuronal NOS (NOS1) in aortic endothelial cells, but no change in endothelial NOS (NOS3) immunoreactivity. However, subendothelial macrophages and smooth muscle showed a different pattern of immunoreactivity of NADPH-diaphorase (NADPH-d), NOS2, ET-1, and NOS1. The lipid rich macrophages in the intima were positively labeled for NADPH-d, NOS1, NOS2, NOS3, and ET-1. Smooth muscle cells in the subendothelium and the medial layers of the vascular wall were also positive for these markers. These results are consistent with the reduction of endothelium-dependent vasorelaxation that is known to occur during the development and progression of atherosclerosis in familial hypercholesterolemia. The data suggest a key role for vasoactive substances in the development of atherosclerosis. PMID- 11502097 TI - The effect of intravenous immunoglobulins on the progression of experimental autoimmune myocarditis in the rat. AB - The aim of this study was to evaluate the efficacy of two experimental regimes of human intravenous immunoglobulins (IVIG) on the progression of experimental autoimmune myocarditis (EAM). EAM is induced by immunization against myosin and represents a T-cell-dependent disorder that progresses toward dilated cardiomyopathy similar to the human equivalent. No effective treatment is currently at hand for management of the disorder, as immunosuppressant drugs are associated with multiple side effects. Three groups of Lewis rats were induced to develop EAM by immunization with porcine myosin and sacrificed 21 days later. Group A received a 5-day regimen of IVIG (800 mg/kg) following induction of the disorder; Group B received a daily dose of IVIG (800 mg/kg) and group C was treated with PBS. IVIG given daily but not during the first 5 days significantly suppressed myocarditis score (0.81 +/- 0.26 and 1.14 +/- 0.42, respectively) in comparison with controls (mean score of 1.78 +/- 0.36). The effect was accompanied by a reduction in the cellular and humoral immune response of the respective animals toward myosin. IVIG was deposited within the extracellular matrix surrounding the damaged myocytes. TNF-alpha expression was reduced in both groups treated with IVIG, whereas iNOS expression paralleled the extent of myocardial inflammation regardless of treatment. IVIG at doses twice those applied for human disease are effective in ameliorating the progression of EAM. The effect may be mediated by suppression of the cellular and humoral response to myosin. IVIG may be found clinically feasible in humans as an adjuvant or single therapy for autoimmune myocarditis. PMID- 11502098 TI - Whole body hyperthermia accelerates atherogenesis in low-density lipoprotein receptor deficient mice. AB - The application of brief periods of heat stress prior to induction of various forms of tissue injury (ischemia-reperfusion, myocardial infarction, endothelial denudation) has been shown to result in preconditioning and attenuation of subsequent damage. Atherosclerosis represents a state of heightened response to injury at the level of the vessel wall, involving endothelial cells, smooth muscle cells, and macrophages. In the current study, we studied the effects of whole body hyperthermia (WBH) on diet-induced atherosclerosis in a murine model. Low-density lipoprotein receptor deficient mice were either exposed to a 30-min WBH (n = 10) or nontreated (n = 7). Animals were given a high-fat ("Paigen"-type) diet to speed the progression of atherosclerosis immediately following WBH for 6 weeks. Aortic and plaque heat shock protein (HSP) 70, suggested to mediate thermotolerance, was assessed by immunohistochemisry and Western blot at different time points following induction of WBH. Aortic sinus plaque formation was significantly accelerated in WBH-treated mice (275,800 +/- 19,540 microm(2) ) in comparison with their control litters (152,100 +/- 18,200 microm(2); P = 0.0004). Plaque composition was also influenced by WBH as lesions were more mature and had an increased proportion of lipid core/fibrous cap accompanied by increased numbers of apoptotic cells. Total cholesterol and triglyceride levels were not affected significantly by WBH. HSP70 protein expression in the aortas was increased 30 min and 6 and 12 h following WBH induction. Thus, induction of WBH, which affords protection in models of arterial injury, appears to have a proatherogenic role in murine atherosclerosis, despite its upregulatory influence on the expression of HSP70. PMID- 11502099 TI - High panel reactive antibody against cross-reactive group antigens as a contraindication to renal allotransplantation. AB - Preformed circulating cytotoxic IgG anti-HLA alloantibodies induced by previous failed grafts, blood transfusion, or pregnancy are a contraindication to allotransplantation and result in hyperacute rejection. These persistent, highly cytotoxic panel reactive antibodies (PRAs) may be specific for epitopes that are shared among HLA antigens known as cross-reactive groups (CREGs). The present investigation includes 24 subjects awaiting renal transplants with flow cytometric PRAs >30%. Eighty-seven percent of the patients developed alloantibodies specific for the mismatched antigens of previous failed grafts. The complement-dependent cytotoxicity test revealed that A1 and A2 antigens were highly immunogenic, whereas A23, B35, and B7 were less so. All patients who formed anti-A1 and anti-A2 also had developed alloantibodies specific for other antigens of the 1C and 2C CREGs, respectively. The presence of anti-class II HLA alloantibodies led to poor graft survival, i.e., a maximum of 2 years. PMID- 11502100 TI - Liver necrosis induced by thyroid hormone administration in rats fed ethanol. AB - Recently it was observed that the hypothyroid state induced by propylthiouracil treatment reduced the elimination rate of ethanol. This was based on the fact that hypothyroid rats could only tolerate 10 g/kg/day of ethanol infusion over 24 h. Euthyroid rats tolerate 13 to 14 g/kg/day of ethanol infusion. In the present report it was postulated that thyroid supplements would increase the daily dose of ethanol that the treated rats could tolerate. Daily thyroid supplements, given intraperitoneally or orally, did, in fact, result in an increase in the elimination rate of ethanol. Death from ethanol overdose did not occur until the amount of ethanol given reached 16 to 19 g/kg/day. In addition, thyroxine supplements blunted or completely inhibited the urinary ethanol cycle. Submassive centrilobular liver necrosis was present in 9 of 10 rats given thyroid hormone and fed ethanol. Periportal duct metaplasia and fibrosis were also observed. Thus, thyroid supplements probably enhanced central hypoxia caused by ethanol to the point where ischemic necrosis developed. The latter healed by scar formation. Thyroid treatment completely prevented ethanol-induced steatohepatitis, indicating that the fatty change is responsive to the metabolic rate. PMID- 11502101 TI - Bax, Bcl-2, and NF-kappaB expression in sanguinarine induced bimodal cell death. AB - The apoptosis related proteins Bax, Bcl-2, and NF-kappaB were analyzed in sanguinarine induced apoptosis and blister cell death (BCD) of K562 erythroleukemia cells and in sanguinarine treated high Bcl-2 expressing JM1 pre-B lymphoblastic cells, utilizing immunofluorescence-flow cytometry. Sanguinarine induced apoptosis of K562 cells was found to have increased Bax expression and decreased NF-kappaB, whereas BCD showed a decrease in Bax expression and an increase in NF-kappaB. In contrast, high Bcl-2 expressing JM1 cells, when exposed to the same concentrations (and duration) of sanguinarine that induced PCD and BCD in K562 cells, failed to show the respective morphologies while showing a concomitant increase in Bcl-2. Results from studies with K562 cells suggest that Bax is pro-apoptotic and also that NF-kappaB activation may be associated with BCD. Results from studies with JM1 cells suggest that Bcl-2 is anti-apoptotic and anti-BCD. Results from JM1 cells strengthen the assumption in the literature of the central role Bcl-2 plays in chemoresistance by assuming an anti-PCD role. These results also suggest that, in JM1 cells, Bcl-2 may further complicate chemoresistance by being anti-BCD in nature, in addition to its anti-PCD role. PMID- 11502102 TI - Interactions between Surfactants and Particles: Dispersion, Surface Modification, and Adsolubilization. AB - Interactions of surfactants and polymers at solid-liquid interfaces can be correlated with many interfacial processes. In this study, we discuss dispersion of particles, surface modification of particles, and adsolubilization using various surfactants and particles. Copyright 2001 Academic Press. PMID- 11502103 TI - Some Thoughts Regarding Theoretical Aspects of the Corrin-Harkins Relation and the Micellization Product of Ionic Micelles. AB - The dependence of the stability of ionic micelles on the ionic strength of the medium is examined analytically without recourse to any explicit expression of the surface potential of micelles. The present study is based on the idea developed by Evans, Mitchell, and Ninham (D. F. Evans, D. J. Mitchell, and B. W. Ninham, J. Phys. Chem. 88, 6344 (1984)) that the interfacial free energy at the water/hydrocarbon core interface is independent of the ionic strength of the medium. The Corrin-Harkins (C-H) relation, a linear relation between the logarithm of the critical micelle concentration (cmc) and the logarithm of the counterion concentration n(C), is obtained in the range of n(C) where the salting out effect is negligible, under the condition that the area per monomer on the micelle surface decreases very weakly with n(C). The "micellization product" of the charged pseudophase model of ionic micelles is discussed. The linear dependence of the surface potential of ionic micelles on n(C) is derived while a part of the effects of salt on the micelle size/shape is allowed. Copyright 2001 Academic Press. PMID- 11502104 TI - Influence of Polydispersity on Adsorption of Nanoparticles. AB - The adsorption of polydisperse, interacting nanoparticles is studied experimentally and discussed in terms of the random sequential adsorption model. Two kinds of polystyrene particles with different size variation (41+/-6 and 107+/-5 nm) were used in adsorption experiments at or close to saturation. The dried monolayer particle films were analyzed with scanning electron microscopy. Selective adsorption of smaller particles resulted in altered size distributions on the surface compared to that in solution. Varying the ionic strength was seen to influence the effective polydispersity of the particles. With increasing salt concentration there was a relative increase in the adsorption of smaller particles, resulting in a large shift toward smaller particle sizes in the size distribution on the surface. Polydispersity gave a slight increase in coverage at high salt concentrations and a decrease in the ordering of the particles on the surface. Copyright 2001 Academic Press. PMID- 11502105 TI - The Effect of Solvent and Fiber Treatment on the Deposition of Organic Silane Solutions Using THF and Acetone. AB - To facilitate the deposition of gamma-ureidopropyltrimethoxysilane monomer onto E glass fibers, anhydrous acetone and THF were used as carrier solvents. Untreated industrial E-glass fibers and cold water plasma treated E-glass fibers were used. Cold water plasma treatment of the fibers increased the hydrophobicity of the surface by increasing the number of hydroxyl groups at the surface; additionally, the amount of aluminium and calcium at the surface was increased. The silane was found to deposit in bilayers with an external surface due to condensed siloxane; aluminium and calcium were also condensed into the network when acetone was used as a deposition solvent. The bilayers were strongly hydrogen bonded via the ureido functional groups. When THF was used as a carrier solvent, a ureido functional surface was obtained due to the THF interfering with the condensation of the silane. The silane was deposited in conical patches of varying heights. Copyright 2001 Academic Press. PMID- 11502106 TI - New Factorial Designs to Evaluate Chemisorption of Divalent Metals on Aminated Silicas. AB - Adsorption processes of Cu(II), Co(II), and Hg(II) on two aminated silica gel surfaces with immobilized ethylenediamine and diethylenediamine groups were studied by a new 2(3) full factorial design. Two metal quantity levels, temperatures of 25 and 50 degrees C, and silica amounts of 100 and 200 mg were employed. Our study indicates that higher factorial design levels increase adsorption for all metals evaluated, with the exception of the mass parameter. Adsorption is also significantly affected by important antagonistic and synergistic effects involving all factors. Both functionalized silica gel surfaces present higher interactions and good perspectives in preconcentration studies for mercury. The factorial design results are also discussed in terms of some solvation properties for each of the metals studied. Copyright 2001 Academic Press. PMID- 11502107 TI - Experimental Study of Fibrin/Fibrin-Specific Molecular Interactions Using a Sphere/Plane Adhesion Model. AB - Fibrin, the biopolymer produced in the final step of the coagulation cascade, is involved in the resistance of arterial thrombi to fragmentation under shear flow. However, the nature and strength of specific interactions between fibrin monomers are unknown. Thus, the shear-induced detachment of spherical monodispersed fibrin coated latex particles in adhesive contact with a plane fibrin-coated glass surface has been experimentally studied, using an especially designed shear stress flow chamber. A complete series of experiments for measuring the shear stress necessary to release individual particles under various conditions (various number of fibrin layers involved in the adhesive contact, absence or presence of plasmin, the main physiological fibrinolytic enzyme) has been performed. The nonspecific DLVO interactions have been shown to be negligible compared to the interactions between fibrin monomers. A simple adhesion model based on the balance of forces and torque on particles, assuming an elastic behavior of the fibrin polymer bonds, to analyze the experimental data in terms of elastic force at rupture of an elementary intermonomeric fibrin bond has been used. The results suggested that this force (of order 400 pN) is an intrinsic quantity, independent of the number of fibrin layers involved in the adhesive contact. Copyright 2001 Academic Press. PMID- 11502108 TI - Colloid Particle Adsorption on Partially Covered (Random) Surfaces. AB - The random sequential adsorption (RSA) approach was used to model irreversible adsorption of colloid particles at surfaces precovered with smaller particles having the same sign of surface charge. Numerical simulations were performed to determine the initial flux of larger particles as a function of surface coverage of smaller particles θ(s) at various size ratios lambda=a(l)/a(s). These numerical results were described by an analytical formula derived from scaled particle theory. Simulations of the long-time adsorption kinetics of larger particles have also been performed. This allowed one to determine upon extrapolation the jamming coverage θ(l)(infinity) as a function of the lambda parameter at fixed smaller particle coverage θ(s). It was found that the jamming coverage θ(l)(infinity) was very sensitive to particle size ratios exceeding 4. Besides yielding θ(l)(infinity), the numerical simulations allowed one to determine the structure of large particle monolayers at the jamming state which deviated significantly from that observed for monodisperse systems. The theoretical predictions suggested that surface heterogeneity, e.g., the presence of smaller sized contaminants or smaller particles invisible under microscope, can be quantitatively characterized by studying larger colloid particle adsorption kinetics and structure of the monolayer. Copyright 2001 Academic Press. PMID- 11502109 TI - Calibration of Polarization-Sensitive and Dual-Angle Laser Light Scattering Methods Using Standard Latex Particles. AB - A polarization-sensitive laser light scattering (PSLLS) method and a dual-angle laser light scattering (DALLS) method have been studied for in situ measurement of submicrometer hydrosol and aerosol particles. By using standard monodisperse polystyrene latex particles suspended in water and air as test particles, calibration of systems built based on the above methods have been performed. The effects of light scattered by agglomerated aerosol particles (multiplets) were corrected by considering the fraction of multiplets as determined with an aerosol measurement technique using a differential mobility analyzer. The change in the measured intensities of scattered light with particle diameter was then determined by calculations based on Mie theory. It was shown that the PSLLS system can determine particle diameters as small as approximately 60 nm for the test hydrosol particles and approximately 100 nm for test aerosol particles, respectively. The DALLS system can determine smaller diameters than the PSLLS system for test particles with no light absorption. The change in scattered light intensities with particle diameter was also investigated by theoretical calculations with various refractive indexes and scattering angles. The PSLLS and DALLS systems promise to become routine measurement tools for absorbing and nonabsorbing particles, respectively. Copyright 2001 Academic Press. PMID- 11502110 TI - Strong and Weak Interactions of Colloidal Particles with High Surface Potentials. AB - A simple method for calculating the interaction force and energy per unit area between two dissimilar plates with high potentials at constant surface potential presented. Using Derjaguin's method and the improved Derjaguin' method, the expressions of the interaction free energy between two dissimilar spheres with high surface potentials are derived. These formulae may be divided into two groups: those for the strong interaction and those for the weak interaction. The juncture of strong and weak interactions is at kappah=4 for dissimilar plates and at kappah=2.8 for spheres The relative error is largest at this point, about 10%. Copyright 2001 Academic Press. PMID- 11502111 TI - Diffusing-Wave Spectroscopy of Concentrated Alumina Suspensions during Gelation. AB - Using diffusing-wave spectroscopy, we followed the aggregation and gelation of concentrated (30 vol%) alumina suspensions. The suspensions were destabilized by either shifting the pH to the isoelectric point or by increasing the ionic strength. Both effects can be achieved continuously and homogeneously by using an enzyme-catalyzed internal chemical reaction. Based on the light-scattering data, we could derive quantitative information about the sol-gel transition and the viscoelastic properties of the gels, as well as a characterization of changes in the microstructure. The elastic moduli determined from light scattering are found to be in good agreement with rheological measurements. Copyright 2001 Academic Press. PMID- 11502112 TI - On the Use of the Hypothesis of Statistically Homogeneous Suspensions in the Calculation of Their Conductivity. AB - In this paper the condition of a "statistically homogeneous" suspension is carefully analyzed. This condition requires that in the presence of a macroscopic gradient of the electric potential there are no macroscopic ion concentration gradients in the system. It is shown that the use of this condition, which should always be required in the calculation of the electrical conductivity of colloidal suspensions, is far from trivial. Furthermore, its hasty use can lead to substantial errors in the calculation of the conductivity increment. Copyright 2001 Academic Press. PMID- 11502113 TI - Kinetics of the Formation of Nano-Sized Platinum Particles in Water-in-Oil Microemulsions. AB - The effect of surfactant type and temperature on the kinetics of the formation of platinum nanoparticles in water-in-oil microemulsions by chemical reduction of PtCl(6)(2-) were examined with time-resolved UV-vis absorption spectroscopy. The surfactants used were poly(ethylene glycol)monododecyl ethers (C(12)E(4), C(12)E(5), C(12)E(6)), sodium bis(2-ethylhexyl)sulphosuccinate (AOT), and mixtures of the alcohol ethoxylates and AOT. The oil domain was n-heptane. The microemulsion droplet size was measured by a dynamic light scattering technique (photon correlation spectroscopy) and the final platinum particle size was determined by transmission electron microscopy. The reaction rate for platinum particle formation was approximately the same in microemulsions based on either of the alcohol ethoxylates but considerably lower for microemulsions based on AOT. In microemulsions based on mixtures of an alcohol ethoxylate and AOT the reaction rate was similar to that obtained when alcohol ethoxylate was the sole surfactant. The reaction was observed to be particularly rapid in microemulsions based on combinations of AOT and C(12)E(5) or C(12)E(6), and the rate was relatively independent of the ratio of the nonionic and anionic surfactants. The reaction was found to be of first order for platinum nanoparticles formed in alcohol ethoxylate-, AOT-C(12)E(5)-, and AOT-C(12)E(6)-based microemulsions, whereas in microemulsions with AOT and AOT-C(12)E(4) the reaction rate seemed to be of higher reaction order. The platinum particles were found to be less than 5 nm in average diameter, which was consistent with the microemulsion droplet size. Copyright 2001 Academic Press. PMID- 11502114 TI - The Electrical Double Layer at Hydrous Manganese Dioxide/Electrolyte Interface. AB - The interfacial properties of two hydrous oxides of manganese, namely an amorphous variety and a poorly crystalline synthetic birnessite, in different electrolyte media, have been studied by using a batch potentiometric titration procedure with a 72-h equilibration period. The data obtained by this procedure are more relevant to a natural water system than those obtained by the rapid titration method as reported earlier [J. Colloid Interface Sci. 131, 103 (1989).]. Although the points of zero charge do not show any major variations, the surface dissociation and complexation (with Na(+) in NaCl) constants differ with the method of titration. Both of the samples are characterized by their high surface charge (low DeltapK(a)) and surface potential, and as a result, complexation takes place possibly within the inner helmholtz plane. It is concluded that the triple-layer model is not applicable to this colloid system and that a basic Stern model would be more suitable in this case. Copyright 2001 Academic Press. PMID- 11502115 TI - Heterogeneity of Ion-Exchange Membranes: The Effects of Membrane Heterogeneity on Transport Properties. AB - The heterogeneity of different cation-exchange membranes (Neosepta CMX, Selemion CMV, and HJC heterogeneous membrane) and their effects on transport properties were investigated using chronopotentiometry, membrane conductivity, and current voltage curves. Modifying the classical Sand equation, a method has been developed to determine the fraction of the conducting region (epsilon) of the ion exchange membrane. The epsilon values of the CMX, CMV, and HJC membranes were 0.93, 0.95, and 0.75, respectively. Considering the characteristics of each membrane-the CMX and CMV are reinforced homogeneous membranes, while the HJC is a heterogeneous membrane-the epsilon values determined in this study seem to be reasonable. The dependence of membrane conductivities and the limiting current densities on the fraction of conducting region of each membrane have also been studied. Copyright 2001 Academic Press. PMID- 11502116 TI - A New Method of Calculating Pore Size Distribution: Analysis of Adsorption Isotherms of N(2) and CCl(4) for a Series of MCM-41 Mesoporous Silicas. AB - The adsorption isotherms of N(2) gas at 77 K and CCl(4) vapor at 283.1(5), 298.1(5), and 308.1(5) K were measured for six samples of the mesoporous silicas having uniform cylindrical pores (MCM-41). The pore radii of the six samples (r(p)), which were evaluated from the alpha(s) plots of the N(2) isotherms, were 1.13, 1.29, 1.50, 1.65, 1.90, and 2.53 nm. The CCl(4) adsorption isotherms show that the capillary condensation occurs at the very narrow P/P(0) range. The core radii of the six adsorbents (r(c)), which were estimated from a comparison plot of the CCl(4) isotherm, were 0.90, 1.01, 1.28, 1.37, 1.60, and 2.17 nm. In the comparison plot, the standard CCl(4) isotherm for nonporous silica was used as the reference isotherm. It has been clarified that the Polanyi adsorption potential of capillary condensation is proportional to the reciprocal of the core radii: RT ln(P(0)/P)=5.37r(c)(-1) nm(-1), ln(P(0)/P)=2.17r(c)(-1) nm(-1) at 298.1(5) K, [A]. The statistical thickness of adsorbed CCl(4) on the curved surface (t((pore))), which was estimated from the difference between the pore radii and the core radii, was given by Eq. [B]: t((pore))=0.188+0.336(P/P(0))+0.382(P/P(0))(2) nm [B], (0.08

25 mol% of pendant octyl groups) resulted in small mixed objects of micellar size (radius about 10 nm). The drop of the mean radius of intermediate structures formed upon the vesicle breakage was also sensitive to temperature. A tentative mechanism was proposed on the basis of kinetics and FFEM studies. Copyright 2001 Academic Press. PMID- 11502121 TI - Thermodynamic Investigation of the Micellization and the Adsorption of Short Perfluoroalkylated Diols via Fluorescence Spectroscopy and Tensiometry. AB - The micellization and adsorption of two short chain perfluorodiols 3,3,4,4,5,5,6,6,6-nonafluorohexane-1,2-diol (nFHD) and 3,3,4,4,5,5,6,6,7,7,8,8,8 tridecafluorooctane-1,2-diol (tFOD) are examined from a thermodynamic point of view as a function of temperature and methanol content. The microenvironment of the fluorinated aggregates is evaluated by the fluorescence probe method using pyrene and a molecular rotor 1,1-dicyano-4-p-dimethylaminophenyl)-1,3-butadiene (DMAPhC). The formation of micellar aggregates being evidenced, the results are discussed in terms of the polarity and of the cohesion behavior of the micellar aggregates by taking into account the methanol (MeOH) effect. The critical micellization concentrations thus determined are compared with those given by surface tension measurements. Micellar and adsorption thermodynamic parameters such as Gibbs free energies, enthalpies, and entropies are determined together with the surface areas. The results are compared with literature data and discussed. A model for describing the adsorption process of the fluorinated compounds upon the influence of methanol is finally proposed. Copyright 2001 Academic Press. PMID- 11502122 TI - Effect of Pentanol Partitioning on Solubilization of Tetrachloroethylene and Gasoline by Sodium Dodecyl Sulfate Micelles. AB - The effect of interfacial pentanol concentrations on solubilization of tetrachloroethylene (PCE) and gasoline by sodium dodecyl sulfate (SDS) micelles was compared to that for dodecane solubilization, which had been measured in a previous study. The solubilization of PCE and gasoline reached their maximum values at a 1 : 3 SDS-to-pentanol molar ratio in the interface. As pentanol concentrations increased beyond that necessary for interfacial saturation, solubilization of PCE and gasoline decreased. This behavior was similar to that observed when dodecane was the oil phase. Electrical conductivity of aqueous SDS/pentanol solutions followed a trend similar to that for oil solubilization, reaching a maximum value at a 1 : 3 molar ratio of SDS to pentanol in the interface. The results of this and previous studies suggest that pentanol partitioning in SDS micelles can be described by a simple two-region model: Region I is the interface between the water-continuous phase and oil and Region II is the micelle inner core. When the mole fraction of pentanol in the interface is less than 0.75, pentanol partitions strongly into Region I, where it acts as a cosurfactant along with SDS and enhances oil solubilization. Above 0.75 mole fraction in the interface, pentanol partitions strongly into Region II, where it acts as a polar oil and competes with other oils for solubilization. Copyright 2001 Academic Press. PMID- 11502123 TI - The Surface Energy and Contact Angles of a Liquid Foam. AB - A liquid foam can be regarded as a single fluid with a characteristic "surface tension". This property is calculated for two- and three-dimensional ordered and disordered foams using a broken bond approach. The contact angle of a foam with a solid or a liquid substrate is also calculated from the energies of the bubbles in contact with the substrate. Experiments were carried out in which the contact angles of foams were measured, with reasonable agreement with the predictions. Copyright 2001 Academic Press. PMID- 11502124 TI - Sugar-Ester Nonionic Microemulsion: Structural Characterization. AB - Surfactants containing sugar components and fatty acids satisfy the quality standards for food application. The food grade sugar ester in this study is a commercial sucrose monoester of stearic acid (abbreviated SES), the oil phase consists of a 1:1 mixture of n-tetradecane and l-butanol. The originally planned food grade oil, a medium chain triglyceride, is substituted by tetradecane because tetradecane is available as a fully deuterated product, which is necessary for some structural investigations. The investigated system is solid at room temperature, but liquefies and structures into a homogeneous microemulsion when heated to above 37 degrees C. The structural characterization of such microemulsions is the aim of this work. The established methods for this purpose are scattering methods, such as small-angle scattering of X-rays and neutrons and dynamic light scattering. These scattering techniques can be used to obtain valuable information on the size, shape, and internal structure of colloids and complex fluids. We started our investigation with the pseudobinary system SES, tetradecane and l-butanol, varying the SES content. The scattering results show that the sugar ester form inverse globular micelles in the oil phase. The size of these micelles is about 6 nm. While the size is nearly constant in a wide SES concentration regime (5 up to 40% surfactant), the volume or aggregation number increases significantly with SES. This is explained by an increasing replacement of l-butanol molecules by sugar-ester molecules in the micelles formed. Moreover, it can be shown that these micelles strongly overlap. Their center-to-center distance is about 3.8 nm at 40% SES at a micellar diameter of 6 nm. The micellar overlap leads to a highly reduced diffusion of the micelles as was found with dynamic light scattering. When incorporating water in the micellar core, the micelles swell up to about 10 nm and the shape of the aggregates becomes more and more elongated with higher water content. Copyright 2001 Academic Press. PMID- 11502125 TI - Wall Slip Phenomena in Oil-in-Water Emulsions: Effect of Some Structural Parameters. AB - The influence of some structural parameters on wall slip phenomena during the flow of oil-in-water model emulsions has been investigated using geometries with smooth and rough surfaces. A wall slip phenomenon has been quantified as the relative difference between the areas under the flow curves obtained using geometries with both types of surfaces. Experimental results demonstrate that wall slip effects phenomena vanish as the disperse phase fraction, the emulsifier content, or the agitation speed applied during the emulsification process increase. These influences have been related to changes in both droplet size and interparticle interactions. In addition to this, the shear stress range in which wall slip is more important coincides with the region at which a shear-induced deflocculation process occurs. In this region, low reproducible values of the flow curve were obtained by using geometries with smooth surfaces. Copyright 2001 Academic Press. PMID- 11502126 TI - The Dynamics of Marangoni-Driven Local Film Drainage between Two Drops. AB - A study of Marangoni-driven local continuous film drainage between two drops induced by an initially nonuniform interfacial distribution of insoluble surfactant is reported. Using the lubrication approximation, a coupled system of fourth-order nonlinear partial differential equations was derived to describe the spatio-temporal evolution of the continuous film thickness and surfactant interfacial concentration. Numerical solutions of these governing equations were obtained using the Numerical Method of Lines with appropriate initial and boundary conditions. A full parametric study was undertaken to explore the effect of the viscosity ratio, background surfactant concentration, the surface Peclet number, and van der Waals interaction forces on the dynamics of the draining film for the case where surfactant is present in trace amounts. Marangoni stresses were found to cause large deformations in the liquid film: Thickening of the film at the surfactant leading edge was accompanied by rapid and severe thinning far upstream. Under certain conditions, this severe thinning leads directly to film rupture due to the influence of van der Waals forces. Time scales for rupture, promoted by Marangoni-driven local film drainage were compared with those associated with the dimpling effect, which accompanies the approach of two drops, and implications of the results of this study on drop coalescence are discussed. Copyright 2001 Academic Press. PMID- 11502127 TI - Effective Compressibility of a Bubbly Slurry. AB - Bubbles trapped within the slurry at the bottom of waste tanks at the Hanford Site expand and contract in response to ambient pressure changes. These bubbles contain hydrogen and other flammable gases that can pose a safety hazard. Determining the effective compressibility of bubbles in the slurry could allow one to make improved estimates of the quantity of gas trapped in the slurry from changes in slurry volume with ambient pressure changes. More broadly, the diffusive growth and mechanical properties of bubbles in a porous medium are important issues in a number of applications. Numerical calculations based on a one-dimensional biconical-pore-network model show that the effective compressibility of a population of bubbles shows hysteresis with pressure increase and decrease. This hysteresis is caused by the sudden jumps of interfaces from pore throat to throat during a pressure decrease and from pore body to body during a pressure increase. Snap-off may occur during these jumps but does not significantly alter the effective compressibility of the population of bubbles. Copyright 2001 Academic Press. PMID- 11502128 TI - Effective Compressibility of a Bubbly Slurry. AB - The goal of this study is to fit model parameters to changes in waste level in response to barometric pressure changes in underground storage tanks at the Hanford Site. This waste compressibility is a measure of the quantity of gas, typically hydrogen and other flammable gases, that can pose a safety hazard, retained in the waste. A one-dimensional biconical-pore-network model for compressibility of a bubbly slurry is presented in a companion paper. Fitting these results to actual waste level changes in the tanks implies that bubbles in the slurry layer are long and the ratio of pore-body radius to pore-throat radius is close to 1; unfortunately, compressibility can not be quantified unambiguously from the data without additional information on pore geometry. Therefore, determining the quantity of gas in the tanks requires more than just waste-level data. The non-uniqueness of the fit is also found with two other simple models: a capillary-tube model with contact angle hysteresis and a spherical-pore model. Copyright 2001 Academic Press. PMID- 11502129 TI - Synthesis and Surface Property of Aqueous Fluorine-Containing Polyurethane. AB - A novel aqueous fluorine-containing polyurethane was prepared with a hydrophobic macromonomer of a perfluoroalkyl group. Two representative properties of the polyurethane, initial particle diameter dispersed in water and surface free energy of coating films, were investigated. The macromonomer was synthesized by radical copolymerization of perfluoroalkylethyl acrylate and methyl methacrylate with a diol of chain-transfer agent in order to attenuate solubility and hydrophobic property. Anionic aqueous polyurethane was obtained with a good hydrophobic film property by one-step condensation polymerization of the macromonomer with hydrophilic comonomers and successive ionization. The polyurethane showed an initial average diameter of less than 1100 nm in water and surface free energies of less than 19 dyn/cm. The water dispersion property and hydrophobic surface property of the polyurethane can be controlled by controlling the content and hydrophobic property of the macromonomer. The incorporation of the macromonomer in the polyurethane backbone did not show a significant effect on the glass transition temperature, or the softness, of the polyurethane. Copyright 2001 Academic Press. PMID- 11502130 TI - Dilute Solution Properties and Aggregation Behavior of Alternate Hetero-arm Copolymer Brushes. AB - Alternate hetero-arm copolymer brushes were synthesized by free-radical copolymerizations of vinylbenzyl-terminated polystyrene macromonomers (PS-VB) with a methacryloyl-terminated poly(ethylene oxide) macromonomer (PEO-MA)/tin tetrachloride (SnCl(4): Lewis acid) complex. Dilute solution properties of such hetero-arm copolymer brushes were investigated by static and dynamic light scattering. Effective diffusion coefficients D(eff) for copolymer brushes possessing large aspect ratios showed almost constant values in the range of polymer concentrations 0-6x10(-3) g/cm(3). These results mean that copolymer brushes formed unimolecular structures even in a good solvent such as benzene. It was speculated from angular dependence measurements that in solution copolymer brushes take geometrically anisotropic conformations such as cylinders. To determine the phase-separated cylindrical domains of alternate copolymer brushes, we constructed large aggregates of copolymer brushes in water. Consequently, alternate copolymer brushes seemed to lead self-assemblies among phase-separated hydrophobic PS domains. Copyright 2001 Academic Press. PMID- 11502131 TI - Electrokinetic Characterization of Poly(N-isopropylacrylamide) Microgel Particles: Effect of Electrolyte Concentration and Temperature. AB - A fundamental study about the characterization of a poly(N-isopropylacrylamide), poly(NIPAM), microgel, cross-linked with N,N'-methylene bisacrylamide (BA), has been carried out. The anionic charge of this system was increased by copolymerization with 2-acrylamido-2methylpropanesulphonic (AMPS) acid. The electrokinetic behavior of these aMPS/NIPAM microgel particles has been investigated. First, the surface charge density was determined by conductimetric titration, and the particle size was obtained by transmission electron microscopy under several conditions. Electrophoretic mobility measurements were carried out as a function of electrolyte concentration and temperature. The hydrodynamic diameter of the particles was also measured by photon correlation spectroscopy as a function of electrolyte concentration and temperature. These size data have been taken into account in order to explain the mobility behavior by using Ohshima's theory for soft particles, with very good agreement between experiments and theoretical predictions. Copyright 2001 Academic Press. PMID- 11502132 TI - Scattering Behavior of Restructured Aggregates: A Simulation Study. AB - Simulations of diffusion-limited cluster-cluster aggregation (DLCA) with no restructuring, full restructuring, and partial restructuring have been performed. The scattering patterns produced from these aggregates have been simulated using the Rayleigh-Gans-Debye approximation. Pure DLCA aggregates produced a scattering pattern with the slope of the fractal region being about -1.8. In contrast, the slope of the fractal region of the scattering pattern for fully restructured aggregates was about -2.1, indicating an increase in fractal dimension. Partial restructuring at large length scales produced an upward turn in the scattering pattern at low qr(o), while at high qr(o) the fractal section of the pure DLCA aggregate was retained. This last result was expected and is consistent with the results and postulations of several other workers. This simulation shows that the type of scattering pattern often obtained from orthokinetic or sheared aggregation can be produced by restructuring of aggregates at large length scales. Copyright 2001 Academic Press. PMID- 11502133 TI - Fiber-like Gold Particles Prepared in Cationic Micelles by UV Irradiation: Effect of Alkyl Chain Length of Cationic Surfactant on Particle Size. AB - Fiber-like gold particles were prepared by irradiation of HAuCl(4) solutions at 253.7 nm in the presence of alkyltrimethylammonium chlorides (C(n)TAC: n=10, 12, 14, and 16). Fiber-like gold particles were obtained above a threshold concentration of C(n)TAC. The length of fiber-like gold particles increases with increasing alkyl chain length of C(n)TAC. Copyright 2001 Academic Press. PMID- 11502134 TI - Rheological Properties of Silica Suspensions in Aqueous Solutions of Block Copolymers and Their Water-Soluble Components. AB - Dynamic moduli of fumed silica suspensions in aqueous solutions of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) block copolymers and PEO homopolymers were measured as a function of surface coverage. Since the block copolymers and PEO are adsorbed on the silica surface through hydrogen bonding between the ether oxygen and the silanol group on the silica surface, the interaction between the silanol groups, which is dominant for the aggregation of silica particles, should be prohibited. Dynamic moduli in the silica suspensions were strongly related to the stability of the silica suspensions and the block copolymer, and the longest PEO portion was useful for stabilizing the silica particles. However, the PEO homopolymer did not support stability of the silica particles, suggesting that chain conformation of the PEO portion in the block copolymer is different from that for the PEO homopolymer. Copyright 2001 Academic Press. PMID- 11502135 TI - Citations and references. PMID- 11502136 TI - Addressing domestic violence through maternity services: policy and practice. AB - OBJECTIVE: to explore current policies and practices in maternity units that aim to identify, assess and support women experiencing domestic violence. DESIGN AND SETTING: a postal survey, conducted between June and October 1999, of all NHS Trusts in England and Wales that provided maternity services. PARTICIPANTS: Heads of Midwifery or the midwife with expertise or interest in domestic violence in each Trust. MAIN OUTCOME MEASURES: use of written policies and agreed practices for identifying and referring women experiencing domestic violence, such as availability of information, routine questioning of all women and offering women an appointment without their partner. RESULTS: 87% (183) of the 211 NHS Trusts providing maternity care participated in the survey. Twelve per cent of units had written policies for identifying women experiencing domestic violence, and a further 30% had some form of agreed practice. Less than half of maternity units routinely offered women an appointment without their partner, and just over half displayed material about domestic violence in places where women receive maternity care. Only three units had undertaken audit on their domestic violence practices. CONCLUSIONS: there is considerable variability around England and Wales in policies and practices related to domestic violence. It is evident that clear guidelines for identification and referral, training, audit and the integration of domestic violence policies with child protection and other policies are necessary to fully address the issues. PMID- 11502137 TI - Ongoing relationships with a personal focus: mothers' perceptions of birth centre versus hospital care. AB - OBJECTIVE: to describe women's perceptions of care in Western Australian birth centres following a previous hospital birth. DESIGN, SETTING AND PARTICIPANTS: an exploratory study was undertaken to examine the care experiences of women from three Western Australian birth centres. Data were obtained from 17 women whose interviews were audio-recorded and transcribed. The research focused upon women's perceptions of their recent birth centre care as compared to previous hospital care during childbirth. FINDINGS: four key themes emerged from the analysis: 'beliefs about pregnancy and birth', 'nature of the care relationship', 'care interactions' and 'care structures'. The themes of 'care interactions' and 'care structures' will be presented in this paper. Care interactions refer to women's opportunities to develop rapport with their carers. Care structures involved the organisational framework in which care was delivered. The first two themes of 'beliefs about pregnancy and birth' and the 'nature of the care relationship' were discussed in a previous paper. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: differences in opportunities for care interactions and care structures were revealed between birth centre and hospital settings. Ongoing, cumulative contacts with midwives in the birth-centre setting were strongly supported by women as encouraging the development of rapport and perception of 'being known' as an individual. Additionally, care structures tailored to women were advocated over the systematised, fragmented care found in hospital settings. PMID- 11502138 TI - Normality and collaboration: mothers' perceptions of birth centre versus hospital care. AB - OBJECTIVE: to describe women's perceptions of care in Western Australian birth centres following a previous hospital birth. DESIGN, SETTING AND PARTICIPANTS: an exploratory design was used to study the care experiences of 17 women recruited from three Western Australian birth centres. Data were obtained from in-depth interviews that explored women's perceptions of their care in both the birth centre and hospital context. FINDINGS: four key themes emerged from the analysis: 'beliefs about pregnancy and birth', 'nature of the care relationship', 'care interactions', and 'care structures'. The themes of 'beliefs about pregnancy and birth' and 'nature of the care relationship' are discussed in this paper. Beliefs about pregnancy and birth refer to the philosophical underpinnings of pregnancy and birth held by women and their carers. Nature of the care relationship identifies women's perceptions of their relationship with health professionals. Care interactions and care structures will be described in a subsequent paper. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: The women's comments suggested differences in philosophy between hospital and birth-centre settings. The philosophy and beliefs of caregivers was an important component of the care experience. Women valued the normality of the birth-centre approach and the opportunity to experience the birth of their child with collaborative support from a midwife. PMID- 11502139 TI - A review of the literature on women's views on their maternity care in the community in the UK. AB - OBJECTIVES: to review the UK literature relating to women's views on community based maternity care between 1970 and 1998. DESIGN: UK research studies examining community-based maternity care were identified by searching several electronic databases using defined search terms. Data extraction was carried out by two or more independent reviewers using a pro-forma. Findings are summarised in this paper. FINDINGS: a total of 624 papers were retrieved and 241 met inclusion criteria. Only 40 papers included some form of comparison group. Examples from more recent and relevant papers focusing on women's views of their maternity care are described and discussed. Women expressed high levels of satisfaction with care. However, there are serious gaps in the research evidence. CONCLUSION AND POLICY IMPLICATIONS: most of the papers included in the review were descriptive and few studies were of a size and quality to enable findings to be generalised to other groups and settings. More information is needed on the care process. There is a need for research studies to examine consumer views where different groups of women experience different care processes. PMID- 11502140 TI - Lesbian mothers' experiences of maternity care in the UK. AB - OBJECTIVE: to explore the maternity care experiences of a sample of lesbians in the UK in order to evaluate service delivery to this group. DESIGN: a descriptive study, using both qualitative and quantitative survey methods to elicit 'rich' accounts of women's experiences. PARTICIPANTS: a convenience sample of 50 women, reporting on a total of 65 pregnancies. FINDINGS: while participants were generally appreciative of the care they received, they also reported high levels of anxiety about the implications of disclosure, together with acute awareness of midwives' personal attitudes and prejudices. Their comments demonstrate the extent to which these issues may negatively impact on quality of care, and the study reveals examples of discomfort, inappropriate service delivery and even hostility. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: 'booking in' and antenatal education are identified as the two areas where service delivery is least effective in meeting the needs of this client group. Findings were used in drawing up the Royal College of Midwives' position paper on the care of lesbian mothers. PMID- 11502141 TI - Women's reluctance to seek help for stress incontinence during pregnancy and following childbirth. AB - OBJECTIVE: to identify whether women were made aware of stress incontinence around the time of childbirth, whether symptomatic women sought help from professionals caring for them at this time, to look at what help they receive in the first instance and to understand why some of those experiencing it chose not to seek help. DESIGN: a qualitative survey was conducted using semi-structured interviews. SETTING: interviews were conducted in participants' own homes. PARTICIPANTS: 42 women who reported symptoms of stress incontinence at eight weeks postpartum, and 15 women who were symptomatic one year following the birth of their baby. FINDINGS: women were reluctant to seek help, although they were often inconvenienced and troubled by the condition. This was predominantly due to the nature of the condition itself, although the relationship with their health care professionals was also a consideration in some cases. The majority of women were not provided with information on the condition but wanted health professionals to provide a warning that the condition could occur. They also wanted health professionals to seek out information about symptoms, rather than the women themselves having to broach the subject. The first line of treatment at this time was usually a recommendation to perform pelvic floor exercises. IMPLICATIONS FOR PRACTICE: health professionals caring for women during pregnancy or following the birth of their baby need to raise awareness of the condition, the treatment available, and to be pro-active in seeking out those experiencing incontinence rather than expecting women to approach them for help. PMID- 11502142 TI - 'Knowing your place': student midwives' views of relationships in midwifery in Ireland. AB - OBJECTIVE: to explore the feelings and views of student midwives of their education as they progressed through their two-year programme in Ireland, with the intention of interpreting and understanding the working and learning world of the participants so that future students might be assisted to improve their educational experiences. DESIGN: phenomenology, using the technique of triangulation in a number of ways. SETTING: all seven midwifery schools in Southern Ireland. PARTICIPANTS: all students in the first intake of 1995 in every midwifery school in Ireland (n=125). DATA COLLECTION: individual and group interviews, diary-keeping and questionnaires. KEY CONCLUSIONS: the findings presented in this paper illustrate the students' views of their relationships with qualified midwives and obstetricians. IMPLICATIONS FOR PRACTICE: an attitude of respect for all students needs to be fostered in hospitals educating midwives if we are to produce caring midwives in the future. The new, extended programme of midwifery education in Ireland should include an emphasis on communication skills and conflict management. A re-organisation of the hierarchical structures evident in midwifery management would lead to an improvement in the experiences of students and childbearing women alike. PMID- 11502143 TI - A comparison of partnership caseload midwifery care with conventional team midwifery care: labour and birth outcomes. AB - OBJECTIVES: to compare the effects of partnership caseload midwifery care, with conventional team midwifery care. Comparisons of labour interventions and birth outcomes were made between the two models of care. DESIGN: a prospective, non randomised clinical trial. SETTING: Women's Hospital at Leicester Royal Infirmary, Leicester, UK in 1998. PARTICIPANTS: 303 women from the experimental group and 308 from the control group (611 in total) matched for age, ethnicity, marital status, parity, gravida and height who gave birth between April 1997 and August 1998. INTERVENTION: the control group received conventional team midwifery care during pregnancy, labour and birth, and the experimental group received care from midwives working in partnerships that provided continuity of care during pregnancy, labour and birth. KEY FINDINGS: 21% of women in the experimental group had an epidural compared with 32% of the controls (OR 0.56 95%, CI 0.39-0.81, P=0.002). The normal vaginal birth rate (74% v 66%, OR 1.45, 95% CI 1.02-2.05, P=0.038), upright birth posture rate (60% v 14%, OR 9.64, 95% CI 5.96-15.61, P= or <0.001), intact perineum rate (40% v 30%, OR 1.57, 95% CI 1.05-2.35, P=0.027), and physiological third stage rate (37% v 1.5%, OR 38.69, 95% CI 11.98-124.89, P= or <0.001) were significantly higher in the experimental group. The induction of labour rate (16% v 23%, OR 0.66, 95% CI 0.44-0.98, P=0.042) was significantly lower in the experimental group. Women in the experimental group had more home births (17% v 1.3%, OR 15.38. 95% CI 5.48-43.14, P= or <0.001); used the midwife led birthing suite more often (28% v 12%, OR 2.77, 95% CI 1.82-4.22, P= or <0.001); were more likely to take an early discharge (two to six hours) from hospital following birth (25% v 3%, OR 11.32. 95% CI 5.55-23.06, P= or <0.001); and were attended in birth more often by either their named midwife (67% v 5%, OR 39.65, 95% CI 22.38-70.25, P= or <0.001) or her partner (known midwife) (84% v 14%, OR 32.74, CI 20.96-51.14, P= or <0.001). IMPLICATIONS FOR PRACTICE: partnership caseload midwifery care resulted in less interventionist labour and more normal birth than conventional team midwifery care. Women in the experimental group had more home births, birth in a midwife-led suite and opted for early discharge home postnatally more often than the controls. They also experienced much higher levels of continuity, particularly of a known midwife during labour and birth. The study findings should encourage other maternity units in the UK to pilot and evaluate the model to see if these benefits are transferable. PMID- 11502145 TI - Hippocampal injections of amyloid beta-peptide 1-40 impair subsequent one trial/day reward learning. AB - The injection of amyloid beta-peptide (Abeta) into rat CNS has been reported to induce cellular neuropathology. The present study investigated whether multiple intrahippocampal injections of Abeta 1-40 would impair one-trial/day reward learning 14 days later. Twenty-four male Sprague-Dawley rats, 3-4 months old, were injected with either Abeta 1-40 or distilled water into seven hippocampal sites bilaterally. Ten rats received 3 nmol Abeta 1-40 in 2 microl of distilled water per injection site, while 14 rats received distilled water alone. Following a 9-day recovery period, rats were gradually food deprived to 82% of their initial body weight. Fourteen days after the intrahippocampal injection, all rats received an initial training trial and three subsequent daily retention trials. Rats receiving Abeta 1-40 were significantly impaired on the second retention trial in terms of accuracy (number of unbaited alleys entered) and on the second and third retention trials in terms of speed (reciprocal of latency to reward). Histological analysis showed that Abeta 1-40 injections produced significant neuronal loss and gliosis. Abeta 1-40 immunoreactivity persisted locally at the injection site and in macrophages 2 weeks following the hippocampal injections. These effects appear to be sequence-specific; rats receiving Abeta 1-42 with a scrambled peptide sequence did not differ significantly from rats receiving distilled water alone in retention of the learning task or degree of histological damage. PMID- 11502146 TI - The effects of genotype, foraging role, and sucrose responsiveness on the tactile learning performance of honey bees (Apis mellifera L.). AB - We analyzed sucrose responsiveness and associative tactile learning in two genetic strains of honey bees under laboratory conditions. These strains differ in their foraging behavior. Bees of the "high" strain preferentially collect pollen. "Low"-strain bees mainly forage for nectar. Responsiveness to different sucrose concentrations and tactile learning were examined using the proboscis extension reflex. Acquisition, extinction of conditioned responses, and responses to an alternative tactile stimulus were tested. High-strain bees are more responsive to sucrose than low-strain bees. Regardless of genotype, pollen foragers are more responsive to sucrose than nectar foragers. In bees of both strains we find the same relationship between responsiveness to sucrose and acquisition. Bees responding to low sucrose concentrations show more often the conditioned response during acquisition than those responding only to higher sucrose concentrations. Extinction of conditioned responses depends on the response probability during acquisition. Discrimination between the two tactile stimuli is affected by genotype but not by responsiveness to sucrose. High-strain bees discriminate better than low-strain bees. Our experiments thus establish links between division of labor, responsiveness to sucrose, and associative learning in honey bees. PMID- 11502147 TI - Fear conditioning-induced alterations of phospholipase C-beta1a protein level and enzyme activity in rat hippocampal formation and medial frontal cortex. AB - We investigated the effects of one-trial fear conditioning on phospholipase C beta1a catalytic activity and protein level in hippocampal formation and medial frontal cortex of untreated control rats and rats prenatally exposed to ethanol. One hour following fear conditioning of untreated control rats, phospholipase C beta1a protein level was increased in the hippocampal cytosolic fraction and decreased in the hippocampal membrane and cortical cytosolic and cortical membrane fractions. Twenty-four hours after fear conditioning, phospholipase C beta1a protein level was reduced in the hippocampal cytosolic fraction and elevated in the cortical nuclear fraction; in addition, 24 h after conditioning, phospholipase C-beta1a activity in the cortical cytosolic fraction was increased. Rats that were exposed prenatally to ethanol displayed attenuated contextual fear conditioning, whereas conditioning to the acoustic-conditioned stimulus was not different from controls. In behavioral control (unconditioned) rats, fetal ethanol exposure was associated with reduced phospholipase C-beta1a enzyme activity in the hippocampal nuclear, cortical cytosolic, and cortical membrane fractions and increased phospholipase C-beta1a protein level in the hippocampal membrane and cortical cytosolic fractions. In certain cases, prenatal ethanol exposure modified the relationship between fear conditioning and changes in phospholipase C-beta1a protein level and/or activity. The majority of these effects occurred 1 h, rather than 24 h, after fear conditioning. Multivariate analysis of variance revealed interactions between fear conditioning, subcellular fraction, and prenatal ethanol exposure for measures of phospholipase C-beta1a protein level in hippocampal formation and phospholipase C-beta1a enzyme activity in medial frontal cortex. In the majority of cases, fear conditioning-induced changes in hippocampal phospholipase C-beta1a protein level were augmented in rats prenatally exposed to ethanol. In contrast, fear conditioning-induced changes in cortical phospholipase C-beta1a activity were, often, in opposite directions in prenatal ethanol-exposed compared to diet control rats. We speculate that alterations in subcellular phospholipase C-beta1a catalytic activity and protein level contribute to contextual fear conditioning and that learning deficits observed in rats exposed prenatally to ethanol result, in part, from dysfunctions in phospholipase C-beta1a signal transduction. PMID- 11502148 TI - Contrasting effects of imidacloprid on habituation in 7- and 8-day-old honeybees (Apis mellifera). AB - We examined the effects of sublethal doses (0.1, 1, and 10 ng per animal) of a new neonicotinoid insecticide, Imidacloprid, on habituation of the proboscis extension reflex (PER) in honeybees (Apis mellifera) reared under laboratory conditions. In untreated honeybees, the habituation of the proboscis extension reflex is age-dependent and there is a significant increase in the number of trials required for habituation in older bees (8-10 days old) as compared to very young bees (4-7 days old). Imidacloprid alters the number of trials needed to habituate the honeybee response to multiple sucrose stimulation. In 7-day-old bees, treatment with Imidacloprid leads to an increase in the number of trials necessary to abolish the response, whereas in 8-day-old bees, it leads to a reduction in the number of trials for habituation (15 min and 1 h after treatment), and to an increase 4 h after treatment. The temporal effects of Imidacloprid in both 7- and 8-day-old bees suggest that 4h after treatment the observed effects are due to a metabolite of Imidacloprid, rather than to Imidacloprid itself. Our results suggest the existence of two distinct subtypes of nicotinic receptors in the honeybee that have different affinities to Imidacloprid and are differentially expressed in 7- and 8-day-old individuals. PMID- 11502149 TI - Use of win-stay and win-shift strategies in place and cue tasks by medial caudate putamen (MCPu) lesioned rats. AB - This study investigated the behavioral function of the medial caudate putamen (MCPu) in the solving of maze tasks. MCPu lesioned rats (n = 35) and control rats (n = 35) were trained for the place or cue task (the four baited arms and four unbaited arms task) in an eight-arm radial maze, which requires the win-stay or the win-shift strategy. In Experiment 1, in which the place task was used, MCPu lesioned rats could learn the task in the win-shift condition, but not in the win stay condition. MCPu lesioned rats made a lot of unbaited errors in the win-stay condition, as they persistently chose adjacent arms. Control rats could learn the tasks in both conditions. In Experiment 2, in which the cue task was used, MCPu lesioned rats and control rats could learn the tasks in both the win-stay and the win-shift conditions. If anything, the performance of MCPu rats was a little better than that of control rats in the win-stay condition. The results of these two experiments revealed that the MCPu was involved in solving the win-stay place task, but not the win-shift place, win-stay cue, and win-shift cue tasks. These findings suggest that the MCPu plays an important role in utilizing both spatial information and switching foraging strategies flexibly and efficiently, that is, processing complicated visuospatial cognition. PMID- 11502150 TI - Effects of a flavor-placement reversal test after different modalities of taste aversion learning. AB - Taste aversion learning is induced through two different behavioral procedures: a short-term or concurrent (two-daily flavors) and a long-term or sequential (one daily flavor) procedure. For the concurrent group of animals, two gustatory/olfactory stimuli are presented separately but at the same time on a daily basis. One is paired with simultaneous intragastric administration of hypertonic NaCl and the other with physiological saline. For the sequential group, the two stimuli are presented on alternate days, one of them followed by intragastric injection of the aversive stimulus and the other by saline, both after a delay of 15 min. The two groups learned the task, but when they were subjected to a flavor-placement reversal test only the sequential group was successful in achieving it. In a second experiment, three groups of animals had to learn concurrent or sequential discrimination tasks (with either simultaneous or delayed administration of the visceral stimulus) using only spatial/proprioceptive cues. The data show that none of the groups learned them under these conditions. The results are discussed in terms of the different modalities of learning. Short-term and long-term taste aversion learning are different in the anatomical structures involved, the number of trials required for acquisition and, as shown in this paper, flexibility. PMID- 11502152 TI - The use and evaluation of primary data in 29 trichloroethylene carcinogen risk assessments. AB - This paper reports the results from a detailed study on how risk assessments of chemicals are actually made. The study is performed by comparing 29 cancer risk assessments made of one and the same chemical substance, namely, trichloroethylene. In this paper, the conclusions that are drawn in these risk assessment documents are described, and differences between the conclusions are explored. This is made within the framework of a proposed cancer risk assessment index. The selection of scientific data for risk assessment purposes is analyzed and the different risk assessors' interpretations and evaluations of individual primary data are compared. It is concluded that the data sets utilized by the trichloroethylene risk assessors are surprisingly incomplete and that biased data selection may have influenced some of the risk assessors' conclusions. Different risk assessors often interpret and evaluate one and the same study in different ways. There are also indications of both interpretation bias and evaluation bias for some of the risk assessors. PMID- 11502153 TI - A review of adverse pregnancy outcomes and formaldehyde exposure in human and animal studies. AB - We examine the potential for reproductive and developmental effects from formaldehyde exposure. Formaldehyde is unlikely to reach the reproductive system in humans in concentrations sufficient to cause damage since it is rapidly metabolized and detoxified upon contact with the respiratory tract. While there are effects seen in in vitro studies or after injection, there is little evidence of reproductive or developmental toxicity in animal studies under exposure levels and routes relevant to humans. Most of the epidemiology studies examined spontaneous abortion and showed some evidence of increased risk (meta-relative risk=1.4, 95% CI 0.9-2.1). We found evidence of reporting biases and publication biases among the epidemiology studies and when these biases were taken into account, we found no evidence of increased risk of spontaneous abortion among workers exposed to formaldehyde (meta-relative risk=0.7, 95% CI 0.5-1.0). The small number of studies on birth defects, low birth weight, and infertility among formaldehyde workers; the limitations in the design of these studies; and the inconsistent findings across these studies make it difficult to draw conclusions from the epidemiology data alone. However, information from experimental studies and studies of metabolism indicate reproductive impacts are unlikely at formaldehyde exposures levels observed in the epidemiology studies. PMID- 11502154 TI - Using dose addition to estimate cumulative risks from exposures to multiple chemicals. AB - The Food Quality Protection Act (FQPA) of 1996 requires the EPA to consider the cumulative risk from exposure to multiple chemicals that have a common mechanism of toxicity. Three methods, hazard index (HI), point-of-departure index (PODI), and toxicity equivalence factor (TEF), have commonly been considered to estimate the cumulative risk. These methods are based on estimates of ED(10) (point of departure) and reference doses from the dose-response functions of individual chemicals. They do not incorporate the actual dose-response function of the mixture from multiple chemical exposures. Dose addition is considered to be an appropriate approach to cumulative risk assessment because it assumes that the chemicals of interest act in accordance with a common mode of action (a similar action). This paper proposes a formal statistical procedure to estimate the cumulative risk by fitting the dose-response model of the mixture under dose addition. The relative potency between two chemicals is estimated directly from the joint dose response model of the mixture. An example data set of four drugs representing four chemicals is used to illustrate the proposed procedure and compare it to the HI, PODI, and TEF methods. PMID- 11502155 TI - Polychlorinated biphenyls, dibenzo-p-dioxins, and dibenzofurans and birth weight and immune and thyroid function in children. AB - None of the publications reviewed provide clinical evidence that PCBs (polychlorinated biphenyls) and related chemicals adversely affect weight, immune, or thyroid function in infants or children born to healthy mothers. Birth weights of term infants fell within the normal range in all studies. The greatest difference between comparison groups was reported by G. G. Fein et al. (1984a, Intrauterine Exposure of Humans to PCBs: Newborn Effects, EPA-600/3-84-060, Environmental Protection Agency; 1984b, J. Pediatr. 105, 315-320). P. R. Taylor et al. (1989, Am. J. Epidemiol. 129, 395-406) reported smaller differences in occupationally exposed women and E. Dar et al. (1992, Environ. Res. 59, 189-201) found that women with higher PCB serum levels had larger babies. S. Patandin et al. (1998, Pediatr Res. 44, 538-545) found a negative association of PCB concentrations in maternal or cord plasma and birth weight of breast and formula fed infants combined but not when breast-fed infants were analyzed separately. L. Rylander et al. (1995, Scand. J. Work Environ. Health 21, 368-375) reported decreased birth weights with higher dietary intake of contaminated fish. Thus, correlations between PCB exposure or polluted fish ingestion and birth weight were inconsistent. Thyroid and immune function were also within the normal range. In none of the papers were normal laboratory reference values provided and overall the statistically significant differences accounted for little of the variance. PMID- 11502156 TI - Pharmacology and toxicology of ethinyl estradiol and norethindrone acetate in experimental animals. AB - For over 30 years various combinations of synthetic estrogens and progestins have been used in oral contraceptive formulations. Ethinyl estradiol (EE) and norethindrone acetate (NA) are common synthetic hormones used in oral contraceptives such as Loestrin, Brevicon, Ortho-Novum, Norlestrin, and Norinyl. In recent years these oral contraceptives have been considered for development in other therapeutic indications. Given the use of these agents for other clinical indications with different and larger target populations, an updated comprehensive review of the toxicology literature of estrogens and progestins is warranted. This review will summarize available data on the pharmacology and toxicology of estrogens and progestins with an emphasis on the specific synthetic hormones EE and NA. Ethinyl estradiol and norethindrone acetate alone or in combination, possess low acute and chronic toxicity. In some studies, EE and/or NA increased the incidence of specific tumors in susceptible strains of rodents and dogs, but not monkeys. These agents are not teratogenic when given in combination. Alone EE and NA have clastogenic properties. Overall, the animal data demonstrates that long-term exposure to EE and NA formulations pose very little health risks to humans. PMID- 11502157 TI - Quantitative cross-species extrapolation in noncancer risk assessment. AB - A procedure has been developed to extrapolate dose-effect findings across species and quantify some of the associated uncertainty via traditional statistical methods. The relationship between dose-effect curves which are known in two species can be described by a k-parameter dose equivalence equation (DEE). When a DEE is determined for as many agents of a given "family" as are known, a k dimensional distribution of DEE parameters would obtain. When an actual extrapolation is to be made for a new agent of the same family which has not been (or cannot be) tested in the species to which results are to be extrapolated, the best estimate of the parameters of the new DEE would be some measure of central tendency and the best estimate of the uncertainty would be the variance/covariance of the k-dimensional distribution of DEE parameters. The method of extrapolation seeks to compliment the mechanistic knowledge or be a substitute for mechanistic methods in the large majority of cases where such understanding is lacking. PMID- 11502158 TI - A biomathematical model of particle clearance and retention in the lungs of coal miners. AB - To understand better the factors influencing the relationships among airborne particle exposure, lung burden, and fibrotic lung disease, we developed a biologically based kinetic model to predict the long-term retention of particles in the lungs of coal miners. This model includes alveolar, interstitial, and hilar lymph node compartments. The 131 miners in this study had worked in the Beckley, West Virginia, area and died during the 1960s. The data used to develop this model include exposure to respirable coal mine dust by intensity and duration within each job, lung and lymph node dust burdens at autopsy, pathological classification of fibrotic lung disease, and smoking history. Initial parameter estimates for this model were based on both human and animal data of particle deposition and clearance and on the biological and physical factors influencing these processes. Parameter estimation and model fit to the data were determined using least squares. Results show that the end-of-life lung dust burdens in these coal miners were substantially higher than expected from first-order clearance kinetics, yet lower than expected from the overloading of alveolar clearance predicted from rodent studies. The best-fitting and most parsimonious model includes processes for first-order alveolar-macrophage mediated clearance and transfer of particles to the lung interstitium. These results are consistent with the particle retention patterns observed previously in the lungs of primates. The findings indicate that rodent models extrapolated to humans, without adjustment for the kinetic differences in particle clearance and retention, would be inadequate for predicting lung dust burdens in humans. Also, this human lung kinetic model predicts greater retained lung dust burdens from occupational exposure than predicted from current human models based on lower exposure data. This model is useful for risk assessment of particle-induced lung diseases, by estimating equivalent internal doses in rodents and humans and predicting lung burdens in humans with occupational dust exposures. PMID- 11502159 TI - A biomathematical model of particle clearance and retention in the lungs of coal miners. II. Evaluation of variability and uncertainty. AB - The objective of this study is to investigate the sources of variability and uncertainty in a previously developed human lung dosimetry model. That three compartment model describes the retention and clearance kinetics of respirable particles in the gas-exchange region of the lungs. It was calibrated using exposure histories and lung dust burden data in U.S. coal miners. A multivariate parameter estimation and optimization method was developed for fitting the dosimetry model to these human data. Models with various assumptions about overloading of alveolar clearance and interstitialization (sequestration) of particles were evaluated. Variability in the estimated clearance rate coefficients was assessed empirically by fitting the model to groups' and to each miner's data. Distributions of lung and lymph node particle burdens were computed at working lifetime exposures, using the variability in the estimated individual clearance rate coefficients. These findings confirm those of the earlier analysis; i.e., the best-fitting exposure-dose model to these data has substantial interstitialization/sequestration of particles and no dose-dependent decline in alveolar clearance. Among miners with different characteristics for smoking, disease, and race, the group median estimated alveolar clearance rate coefficients varied by a factor of approximately 4. Adjustment for these group differences provided some improvement in the dosimetry model fit to all miners (up to 25% reduction in MSE), although unexplained interindividual differences made up the largest source of variability. The predicted mean lung and lymph node particle burdens at age 75 after exposure to respirable coal mine dust at 2 mg/m(2) for a 45-year working lifetime were 12 g (5th and 95th percentiles, 3.0 26 g) and 1.9 g (0.26-5.3), respectively. This study provides quantitative information on variability in particle retention and clearance kinetics in humans. It is useful for risk assessment by providing estimated lung dust burdens associated with occupational exposure to respirable particles. PMID- 11502161 TI - Dendritic cell migration to lymph nodes: cytokines, chemokines, and lipid mediators. AB - Mobilization of dendritic cells into lymphatic vessels requires cytokine stimulation and induction of the chemokine receptor CCR7. The respective roles of the CCR7 ligands CCL19 and CCL21 in mediating migration are not fully defined, but chemotaxis to CCL19 mediates Langerhans cell exit from the epidermis. Optimal chemotaxis to CCL19 occurs when DCs are triggered with exogenous leukotriene C(4), an eicosanoid transported out of the cell via the ATP binding cassette (ABC) transporter multidrug resistance related protein 1 (MRP1, ABCC1). Indeed, MRP1 and the related multidrug resistance protein 1 (MDR1, p-glycoprotein, ABCB1) may control the intracellular and extracellular accumulation of key signaling lipids that regulate dendritic cell migration. PMID- 11502162 TI - Dendritic cell subsets and the regulation of Th1/Th2 responses. AB - Cells of the dendritic family are suited to perform two distinct functions at two discrete locations. In the peripheral tissues, dendritic cells (DC) act as sentinels for "dangerous" antigens. They then migrate into the lymphoid organ, where they initiate activation of T lymphocytes which are specific for these antigens. During their migration, DC shift from an antigen-capturing mode to a T cell sensitizing mode. In addition to switching on the immune response, subtypes of DC appear to influence the character of T cell differentiation, i.e. the Th1/Th2 balance. We will review the cellular and molecular bases of Th1-Th2 development by DC subsets, and will focus primarily, although not exclusively, on mouse DC. PMID- 11502163 TI - Plasmacytoid monocytes/T cells: a dendritic cell lineage? AB - Plasmacytoid monocytes/T cells were first described in 1958, yet their origin and function have remained enigmatic. Recently a series of publications brought these cells to the forefront of immunological research. Indeed, plasmacytoid monocytes/T-cells contain natural type-I interferon producing cells and can differentiate in vitro into dendritic cells (DC). It has been proposed that plasmacytoid monocytes/T-cells represent a distinct lineage of cells whose fate it is to differentiate into dendritic cells. Herein we will review recent advances in our understanding of plasmacytoid monocytes/T cells and highlight arguments in favor or against this lineage hypothesis. We propose that plasmacytoid monocytes/T cells represent a composite group of both myeloid and lymphoid early-committed cells that are characterized by their ability to differentiate in vitro into DC. PMID- 11502164 TI - Dendritic cells and tumor immunity. AB - Researchers and clinicians have tried for decades to use the mechanisms of immunity for the fight against cancer. Early attempts aimed at the instrumentation of soluble immune mediators such as antibodies or cytotoxic proteins for the therapy of malignancies. Major improvements in understanding the induction and regulation of cellular immunity have now made it possible to generate effector cells in cancer patients which are specific for the neoplastic disease. At the beginning of every cellular immune reaction against cancers tumor antigens have to be presented to T cells in order to activate them and drive them into clonal expansion. This is done by antigen presenting cells, the most powerful of which is the dendritic cell (DC). While DC were hard to isolate initially, they can be generated in large numbers in vitro today and manipulated in multiple ways before given back to a patient to induce tumor immunity. Thus, a great amount of hope lies in the use of DC as inducers of tumor immunity. However, the first clinical studies, which have now been completed with only limited success make clear, that still a lot of open questions remain to be answered. This review tries to give an overview of this rapidly developing field, mentioning the major conceptual approaches and techniques, but also discussing important caveats. The next years will show whether we can improve our understanding of DC biology and the mechanisms of immune induction strongly enough to effectively employ DC for immunotherapy of cancer. PMID- 11502165 TI - Dendritic cells and viral immunity: friends or foes? AB - Dendritic cells (DC) residing in epithelial tissues of various mucosae and the skin are characterized by the unique ability to capture antigens and migrate to draining lymph nodes, where they can activate naive and memory T cells. Although DC play a pivotal role in inducing protective immunity to viral infection, they can also be exploited by viruses to evade the host immune response, induce immune suppression, or serve as latent viral reservoirs. Thus, virus interactions with DC may lead to an immune response that can be protective, but does not necessarily lead to complete virus elimination, resulting in immunopathology. PMID- 11502166 TI - DCs and peripheral T cell tolerance. AB - Tolerance is a state in which the immune system as a whole fails to make an active response to antigen. Three mutually exclusive mechanisms appear to account for the fate of antigen-specific peripheral T cells within a tolerant animal: maintenance of naive status, deletion after responding to antigen, and long term survival after responding to antigen, a mechanism that should probably be considered part of the spectrum of memory responses. The types and functional status of the DCs that present antigen in each case remain controversial. This review will summarize the indirect evidence that underlies some of the hypotheses that account for peripheral T cell tolerance. PMID- 11502167 TI - Potential of tolerogenic dendritic cells for transplantation. AB - Dendritic cells (DC) are professional antigen (Ag)-presenting cells considered traditionally as the passenger leukocytes that, after migration from transplanted tissues, stimulate allospecific naive T cell responses and trigger acute rejection. However, there is recent evidence that, besides their role in central T lymphocyte deletion in the thymus, DC perform a crucial function to induce/maintain peripheral T cell tolerance. This paper outlines conceptual models that try to explain how DC may induce/maintain tolerance. It also considers how such ideas have been implemented recently in an effort to generate tolerogenic DC to induce donor Ag-specific tolerance/ immunosuppression and prolonged allograft survival. These approaches include genetic engineering of donor- or recipient-derived DC to express molecules capable of promoting tolerance to alloAg. PMID- 11502168 TI - Probing inhibitors binding to human urokinase crystals by Raman microscopy: implications for compound screening. AB - Inhibition of urokinase activity represents a promising target for antimetastatic therapy for several types of tumor. The present study sets out to investigate the potential of Raman spectroscopy for defining the molecular details of inhibitor binding to this enzyme, with emphasis on single crystal studies. It is demonstrated that high quality Raman spectra from a series of five inhibitors bound individually to the active site of human urokinase can be obtained in situ from urokinase single crystals in hanging drops by using a Raman microscope. After recording the spectrum of the free crystal, a solution of inhibitor containing an amidine functional group on a naphthalene ring was added, and the spectrum of the crystal-inhibitor complex was obtained. The resulting difference Raman spectrum contained only vibrational modes due to bound inhibitor, originating from the protonated group, i.e., the amidinium moiety, as well as naphthalene ring modes and features from other functionalities that made up each inhibitor. The identification of the amidinium modes was placed on a quantitative basis by experimental and theoretical work on naphthamidine compounds. For the protonated group, -C-(NH2)(2)(+), the symmetric stretch occurs near 1520 cm(-1), and a less intense antisymmetric mode appears in the Raman spectra near 1680 cm( 1). The presence of vibrational modes near 1520 cm(-1) in each of the Raman difference spectra of the five complexes examined unambiguously identifies the protonated form of the amidinium group in the active site. Several advantages were found for single crystal experiments over solution studies of inhibitor enzyme complexes, and these are discussed. The use of single crystals permits competitive binding experiments that cannot be undertaken in solution in any kind of homogeneous assay format. The Raman difference spectrum for a single crystal that had been exposed to equimolar amounts of all five inhibitors in the hanging drop showed only the Raman signature of the compound with the lowest K(i). These findings suggest that the Raman approach may offer a route in the screening of compounds in drug design applications as well as an adjunct to crystallographic analysis. PMID- 11502169 TI - Yeast mitochondrial dehydrogenases are associated in a supramolecular complex. AB - Separation of yeast mitochondrial complexes by colorless native polyacrylamide gel electrophoresis led to the identification of a supramolecular structure exhibiting NADH-dehydrogenase activity. Components of this complex were identified by N-terminal Edman degradation and matrix-assisted laser desorption ionization mass spectrometry. The complex was found to contain the five known intermembrane space-facing dehydrogenases, namely two external NADH dehydrogenases Nde1p and Nde2p, glycerol-3-phosphate dehydrogenase Gut2p, D- and L-lactate-dehydrogenases Dld1p and Cyb2p, the matrix-facing NADH-dehydrogenase Ndi1p, two probable flavoproteins YOR356Wp and YPR004Cp, four tricarboxylic acids cycle enzymes (malate dehydrogenase Mdh1p, citrate synthase Cit1p, succinate dehydrogenase Sdh1p, and fumarate hydratase Fum1p), and the acetaldehyde dehydrogenase Ald4p. The association of these proteins is discussed in terms of NADH-channeling. PMID- 11502170 TI - Recognition of ATGA sequences by the unfused aromatic dication DB293 forming stacked dimers in the DNA minor groove. AB - Furamidine and related diamidines represent a promising series of drugs active against widespread parasites, in particular the Pneumocystic carinii pathogen. In this series, the phenylfuranbenzimidazole diamidine derivative DB293 was recently identified as the first unfused aromatic dication capable of forming stacked dimers in the DNA minor groove of GC-containing sequences. Here we present a detailed biochemical and biophysical characterization of the DNA sequence recognition properties of DB293. Three complementary footprinting techniques using DNase I, Fe(II)-EDTA, and an anthraquinone photonuclease were employed to locate binding sites for DB293 in different DNA restriction fragments. Two categories of sites were identified by DNase I footprinting: (i) 4/5 bp sequences containing contiguous A.T pairs, such as 5'-AAAA and 5'-ATTA; and (ii) sequences including the motif 5'-ATGA.5'-TCAT. In particular, a 13-bp sequence including two contiguous ATGA motifs provided a highly preferential recognition site for DB293. Quantitative footprinting analysis revealed better occupancy of the 5' ATGA site compared to the AT-rich sites. Preferential binding of DB293 to ATGA sites was also observed with other DNA fragments and was confirmed independently by means of hydroxyl radical footprinting generated by the Fe(II)-EDTA system, as well as by a photofootprinting approach using the probe anthraquinone-2-sulfonate (AQS). In addition, this photosensitive reagent revealed the presence of sites of enhanced cutting specific to DB293. This molecule, but not other minor groove binders such as netropsin, induces specific local structural changes in DNA near certain binding sites, as independently shown by DNase I and the AQS probe. Recognition of the ATGA sequence by DB293 was investigated further using melting temperature experiments and surface plasmon resonance (SPR). The use of different hairpin oligonucleotides showed that DB293 can interact with AT sites via the formation of 1:1 drug-DNA complexes but binds much more strongly, and cooperatively, to ATGA-containing sequences to form 2:1 drug-DNA complexes. DB293 binds strongly to ATGA sequences with no significant context dependence but is highly sensitive to the orientation of the target sequence. The formation of 2:1 DB293/DNA complexes is abolished by reversing the sequence 5'-ATGA-->3'-ATGA, indicating that directionality plays an important role in the drug-DNA recognition process. Similarly, a single mutation in the A[T-->G]GA sequence is very detrimental to the dimer interactions of DB293. From the complementary footprinting and SPR data, the 5'-ATGA sequence is identified as being a highly favored dimer binding site for DB293. The data provide clues for delineating a recognition code for diamidine-type minor groove binding agents, and ultimately to guide the rational design of gene regulatory molecules targeted to specific sites of the genetic material. PMID- 11502171 TI - The nonmutagenic (R)- and (S)-beta-(N(6)-adenyl)styrene oxide adducts are oriented in the major groove and show little perturbation to DNA structure. AB - Conformations of (R)-beta-(N(6)-adenyl)styrene oxide and (S)-beta-(N(6) adenyl)styrene oxide adducts at position X(6) in d(CGGACXAGAAG).d(CTTCTTGTCCG), incorporating codons 60, 61 (underlined), and 62 of the human N-ras protooncogene, were refined from (1)H NMR data. These were designated as the beta R(61,2) and beta-S(61,2) adducts. A total of 533 distance restraints and 162 dihedral restraints were used for the molecular dynamics calculations of the beta S(61,2) adduct, while 518 distances and 163 dihedrals were used for the beta R(61,2) adduct. The increased tether length of the beta-adducts results in two significant changes in adduct structure as compared to the corresponding alpha styrenyl adducts [Stone, M. P., and Feng, B. (1996) Magn. Reson. Chem. 34, S105 S114]. First, it reduces the distortion introduced into the DNA duplex. For both the beta-R(61,2) and beta-S(61,2) adducts, the styrenyl moiety was positioned in the major groove of the duplex with little steric hindrance. Second, it mutes the influence of stereochemistry at the alpha-carbon such that both the beta-R(61,2) and beta-S(61,2) adducts exhibit similar conformations. The results were correlated with site-specific mutagenesis experiments that revealed the beta R(61,2) and beta-S(61,2) adducts were not mutagenic and did not block polymerase bypass. PMID- 11502172 TI - A structural model for the ternary cleavable complex formed between human topoisomerase I, DNA, and camptothecin. AB - Using the X-ray crystal structure of the human topoisomerase I (TOP1)-DNA cleavable complex, we have developed a general model for the ternary drug-DNA TOP1 cleavable complex formed with camptothecin (CPT) and its analogues. This model has the drug intercalated between the -1 and +1 base pairs, with the E-ring pointing into the minor groove and the A-ring directed toward the major groove. The ternary complex is stabilized by an array of hydrogen bonding and hydrophobic interactions between the drug and both the enzyme and the DNA. Significantly, the proposed model is consistent with the current body of experimental mutation, cross-linking, and structure-activity data. In addition, the model reveals potential sites of interaction that can provide a rational basis for the design of next generation compounds as well as for de novo drug design. PMID- 11502173 TI - Site-directed mutagenesis and X-ray crystallography of the PQQ-containing quinoprotein methanol dehydrogenase and its electron acceptor, cytochrome c(L). AB - Two proteins specifically involved in methanol oxidation in the methylotrophic bacterium Methylobacterium extorquens have been modified by site-directed mutagenesis. Mutation of the proposed active site base (Asp303) to glutamate in methanol dehydrogenase (MDH) gave an active enzyme (D303E-MDH) with a greatly reduced affinity for substrate and with a lower activation energy. Results of kinetic and deuterium isotope studies showed that the essential mechanism in the mutant protein was unchanged, and that the step requiring activation by ammonia remained rate limiting. No spectrally detectable intermediates could be observed during the reaction. The X-ray structure, determined to 3 A resolution, of D303E MDH showed that the position and coordination geometry of the Ca2+ ion in the active site was altered; the larger Glu303 side chain was coordinated to the Ca2+ ion and also hydrogen bonded to the O5 atom of pyrroloquinoline quinone (PQQ). The properties and structure of the D303E-MDH are consistent with the previous proposal that the reaction in MDH is initiated by proton abstraction involving Asp303, and that the mechanism involves a direct hydride transfer reaction. Mutation of the two adjacent cysteine residues that make up the novel disulfide ring in the active site of MDH led to an inactive enzyme, confirming the essential role of this remarkable ring structure. Mutations of cytochrome c(L), which is the electron acceptor from MDH was used to identify Met109 as the sixth ligand to the heme. PMID- 11502174 TI - Reactions of dimethylsulfoxide reductase in the presence of dimethyl sulfide and the structure of the dimethyl sulfide-modified enzyme. AB - The bis-molybdopterin enzyme dimethylsulfoxide reductase (DMSOR) from Rhodobacter capsulatus catalyzes the conversion of dimethyl sulfoxide (DMSO) to dimethyl sulfide (DMS), reversibly, in the presence of suitable e(-)-donors or e(-) acceptors. The catalytically significant intermediate formed by reaction of DMSOR with DMS ('the DMS species') and a damaged enzyme form derived by reaction of the latter with O(2) (DMS-modified enzyme, DMSOR(mod)D) have been investigated. Evidence is presented that Mo in the DMS species is not, as widely assumed, Mo(IV). Formation of the DMS species is reversed on removing DMS or by addition of an excess of DMSO. Equilibrium constants for the competing reactions of DMS and DMSO with the oxidized enzyme (K(d) = 0.07 +/- 0.01 and 21 +/- 5 mM, respectively) that control these processes indicate formation of the DMS species occurs at a redox potential that is 80 mV higher than that required, according to the literature, for reduction of Mo(VI) to Mo(IV) in the free enzyme. Specificity studies show that with dimethyl selenide, DMSOR yields a species analogous to the DMS species but with the 550 nm peak blue-shifted by 27 nm. It is concluded from published redox potential data that this band is due to metal-to-ligand charge transfer from Mo(V) to the chalcogenide. Since the DMS species gives no EPR signal in the normal or parallel mode, a free radical is presumed to be in close proximity to the metal, most likely on the S. The species is thus formulated as Mo(V)-O-S(*)Me(2). Existing X-ray crystallographic and Raman data are consistent with this structure. Furthermore, 1e(-) oxidation of the DMS species with phenazine ethosulfate yields a Mo(V) form without an -OH ligand, since its EPR signal shows no proton splittings. This form presumably arises via dissociation of DMSO. The structure of DMSOR(mod)D has been determined by X-ray crystallography. All four thiolate ligands and Ogamma of serine-147 remain coordinated to Mo, but there are no terminal oxygen ligands and Mo is Mo(VI). Thus, it is a dead-end species, neither oxo group acceptance nor e(-)-donation being possible. O(2)-dependent formation of DMSOR(mod)D represents noncatalytic breakdown of the DMS species by a pathway alternative to that in turnover, with oxidation to Mo(VI) presumably preceding product release. Steps in the forward and backward catalytic cycles are discussed in relation to earlier stopped-flow data. The finding that in the back-assay the Mo(IV) state may at least in part be by-passed via two successive 1e(-) reactions of the DMS species with the e(-) acceptor, may have implications in relation to the existence of separate molybdopterin enzymes catalyzing DMSO reduction and DMS oxidation, respectively. PMID- 11502175 TI - Human ferrochelatase: characterization of substrate-iron binding and proton abstracting residues. AB - The terminal step in heme biosynthesis, the insertion of ferrous iron into protoporphyrin IX to form protoheme, is catalyzed by the enzyme ferrochelatase (EC 4.99.1.1). A number of highly conserved residues identified from the crystal structure of human ferrochelatase as being in the active site were examined by site-directed mutagenesis. The mutants Y123F, Y165F, Y191H, and R164L each had an increased K(m) for iron without an altered K(m) for porphyrin. The double mutant R164L/Y165F had a 6-fold increased K(m) for iron and a 10-fold decreased V(max). The double mutant Y123F/Y191F had low activity with an elevated K(m) for iron, and Y123F/Y165F had no measurable activity. The mutants H263A/C/N, D340N, E343Q, E343H, and E343K had no measurable enzyme activity, while E343D, E347Q, and H341C had decreased V(max)s without significant alteration of the K(m)s for either substrate. D340E had near-normal kinetic parameters, while D383A and H231A had increased K(m)s for iron. On the basis of these data and the crystal structure of human ferrochelatase, it is proposed that residues E343, H341, and D340 form a conduit from H263 in the active site to the protein exterior and function in proton extraction from the porphyrin macrocycle. The role of H263 as the porphyrin proton-accepting residue is central to catalysis since metalation only occurs in conjunction with proton abstraction. It is suggested that iron is transported from the exterior of the enzyme at D383/H231 via residues W227 and Y191 to the site of metalation at residues R164 and Y165 which are on the opposite side of the active site pocket from H263. This model should be general for mitochondrial membrane-associated eucaryotic ferrochelatases but may differ for bacterial ferrochelatases since the spatial orientation of the enzyme within prokaryotic cells may differ. PMID- 11502176 TI - A novel family of hairpin peptides that inhibit IgE activity by binding to the high-affinity IgE receptor. AB - A family of structured peptides that bind to FcepsilonRIalpha, the alpha-chain of the high-affinity receptor for IgE, has been identified. Binding selections using FcepsilonRIalpha and polyvalent peptide-phage libraries yielded a dominant 18 residue peptide-phage clone, as well as related sequences that did not resemble fragments of IgE. Synthetic peptides based on these sequences inhibited IgE binding to its receptor, and were found by NMR analysis to adopt a stable beta hairpin structure in solution. Optimized peptides with micromolar receptor affinity exhibited high stability in biological fluids and inhibited cellular histamine release in an in vitro bioassay of IgE activity. The structure-activity relationships of these peptides, which are less than 1% of the size of IgE, suggest an overlap between their binding site and that of IgE on FcepsilonRI. Thus, the peptides demonstrate that blocking a small epitope on this receptor chain is sufficient to block IgE activity. Such structured peptides represent a possible starting point for the design of novel antagonists, and offer the potential for testing in vivo a new approach for treating allergic disease. PMID- 11502177 TI - beta-Ketoacyl-[acyl carrier protein] synthase I of Escherichia coli: aspects of the condensation mechanism revealed by analyses of mutations in the active site pocket. AB - beta-Ketoacyl-[acyl carrier protein (ACP)] synthase forms new carbon-carbon bonds in three steps: transfer of an acyl primer from ACP to the enzyme, decarboxylation of the elongating substrate and its condensation with the acyl primer substrate. Six residues of Escherichia coli beta-ketoacyl-ACP synthase I (KAS I) implicated in these reactions were subjected to site-directed mutagenesis. Analyses of the abilities of C163A, C163S, H298A, D306A, E309A, K328A, and H333A to carry out the three reactions lead to the following conclusions. The active site Cys-163 is not required for decarboxylation, whereas His-298 and His-333 are indispensable. Neither of the histidines is essential for increasing the nucleophilicity of Cys-163 to enable transfer of the acyl primer substrate. Maintenance of the structural integrity of the active site by Asp-306 and Glu-309 is required for decarboxylation but not for transfer. One function of Lys-328 occurs very early in catalysis, potentially before transfer. These results in conjunction with structural analyses of substrate complexes have led to a model for KAS I catalysis [Olsen, J. G., Kadziola, A., von Wettstein Knowles, P., Siggaard-Andersen, M., and Larsen, S. (2001) Structure 9, 233-243]. Another facet of catalysis revealed by the mutant analyses is that the acyl primer transfer activity of beta-ketoacyl-ACP synthase I is inhibited by free ACP at physiological concentrations. Differences in the inhibitory response by individual mutant proteins indicate that interaction of free ACP with Cys-163, Asp-306, Glu-309, Lys-328, and His-333 might form a sensitive regulatory mechanism for the transfer of acyl primers. PMID- 11502178 TI - Backbone dynamics in dihydrofolate reductase complexes: role of loop flexibility in the catalytic mechanism. AB - To elucidate the influence of local motion of the polypeptide chain on the catalytic mechanism of an enzyme, we have measured (15)N relaxation data for Escherichia coli dihydrofolate reductase in three different complexes, representing different stages in the catalytic cycle of the enzyme. NMR relaxation data were analyzed by the model-free approach, corrected for rotational anisotropy, to provide insights into the backbone dynamics. There are significant differences in the backbone dynamics in the different complexes. Complexes in which the cofactor binding site is occluded by the Met20 loop display large amplitude motions on the picosecond/nanosecond time scale for residues in the Met20 loop, the adjacent betaF-betaG loop and for residues 67-69 in the adenosine binding loop. Formation of the closed Met20 loop conformation in the ternary complex with folate and NADP(+), results in attenuation of the motions in the Met20 loop and the betaF-betaG loop but leads to increased flexibility in the adenosine binding loop. New fluctuations on a microsecond/millisecond time scale are observed in the closed E:folate:NADP(+) complex in regions that form hydrogen bonds between the Met20 and the betaF-betaG loops. The data provide insights into the changes in backbone dynamics during the catalytic cycle and point to an important role of the Met20 and betaF-betaG loops in controlling access to the active site. The high flexibility of these loops in the occluded conformation is expected to promote tetrahydrofolate-assisted product release and facilitate binding of the nicotinamide ring to form the Michaelis complex. The backbone fluctuations in the Met20 loop become attenuated once it closes over the active site, thereby stabilizing the nicotinamide ring in a geometry conducive to hydride transfer. Finally, the relaxation data provide evidence for long-range motional coupling between the adenosine binding loop and distant regions of the protein. PMID- 11502179 TI - Purification and characterization of a soluble bioactive amino-terminal extracellular domain of the human thyrotropin receptor. AB - The amino-terminal ectodomain of the human TSH receptor has been expressed at the surface of CHO cells as a glycosylphosphatidylinositol-anchored molecule containing a 10-residue histidine tag close to its C terminus. The soluble ectodomain could be released from the cells by treatment with a glycosylphosphatidylinositol-phospholipase C and purified to apparent homogeneity by cobalt-Sepharose chromatography. Two nanomoles of material was obtained, which was suitable for analysis by mass spectrometry. This allowed the identification of four out of the six potential N-glycosylation sites as being effectively glycosylated. A proportion of the purified soluble ectodomain displayed specific binding of (125)I-labeled TSH, allowing for the first time performance of classical saturation binding experiments. Two classes of high-affinity binding sites were identified: site A, K(d) 0.014 nM; site B, K(d) 0.83 nM. The significance of site A, whose affinity is much higher than for the holoreceptor at the surface of intact cells, remains to be clarified. The purified ectodomain was capable of inhibiting efficiently the thyroid stimulating activity of immunoglobulins from patients with Graves' disease. It allowed computation of the amounts of these immunoglobulins in patient's serum, giving values up to 10 microg/mL. Contrary to all currently available assays, the soluble ectodomain of the TSH receptor purified in a functionally competent conformation allows direct studies of its interactions with TSH and autoantibodies and opens the way to structural studies. PMID- 11502180 TI - Structure of an active soluble mutant of the membrane-associated (S)-mandelate dehydrogenase. AB - The structure of an active mutant of (S)-mandelate dehydrogenase (MDH-GOX2) from Pseudomonas putida has been determined at 2.15 A resolution. The membrane associated flavoenzyme (S)-mandelate dehydrogenase (MDH) catalyzes the oxidation of (S)-mandelate to give a flavin hydroquinone intermediate which is subsequently reoxidized by an organic oxidant residing in the membrane. The enzyme was rendered soluble by replacing its 39-residue membrane-binding peptide segment with a corresponding 20-residue segment from its soluble homologue, glycolate oxidase (GOX). Because of their amphipathic nature and peculiar solubilization properties, membrane proteins are notoriously difficult to crystallize, yet represent a large fraction of the proteins encoded by genomes currently being deciphered. Here we present the first report of such a structure in which an internal membrane-binding segment has been replaced, leading to successful crystallization of the fully active enzyme in the absence of detergents. This approach may have general application to other membrane-bound proteins. The overall fold of the molecule is that of a TIM barrel, and it forms a tight tetramer within the crystal lattice that has circular 4-fold symmetry. The structure of MDH-GOX2 reveals how this molecule can interact with a membrane, although it is limited by the absence of a membrane-binding segment. MDH-GOX2 and GOX adopt similar conformations, yet they retain features characteristic of membrane and globular proteins, respectively. MDH-GOX2 has a distinctly electropositive surface capable of interacting with the membrane, while the opposite surface is largely electronegative. GOX shows no such pattern. MDH appears to form a new class of monotopic integral membrane protein that interacts with the membrane through coplanar electrostatic binding surfaces and hydrophobic interactions, thus combining features of both the prostaglandin synthase/squaline hopine cyclase and the C-2 coagulation factor domain classes of membrane proteins. PMID- 11502181 TI - Structural characterization of a six-nucleotide RNA hairpin loop found in Escherichia coli, r(UUAAGU). AB - The binding region of the Escherichia coli S2 ribosomal protein contains a conserved UUAAGU hairpin loop. The structure of the hairpin formed by the oligomer r(GCGU4U5A6A7G8U9CGCA), which has an r(UUAAGU) hairpin loop, was determined by NMR and molecular modeling techniques as part of a study aimed at characterizing the structure and thermodynamics of RNA hairpin loops. Thermodynamic data obtained from melting curves for this RNA oligomer show that it forms a hairpin in solution with the following parameters: DeltaH degrees = 42.8 +/- 2.2 kcal/mol, DeltaS degrees = -127.6 +/- 6.5 eu, and DeltaG degrees (37) = -3.3 +/- 0.2 kcal/mol. Two-dimensional NOESY WATERGATE spectra show an NOE between U imino protons, which suggests that U4 and U9 form a hydrogen bonded U.U pair. The U5(H2') proton shows NOEs to both the A6(H8) proton and the A7(H8) proton, which is consistent with formation of a "U" turn between nucleotides U5 and A6. An NOE between the A7(H2) proton and the U9(H4') proton shows the proximity of the A7 base to the U9 sugar, which is consistent with the structure determined for the six-nucleotide loop. In addition to having a hydrogen-bonded U.U pair as the first mismatch and a U turn, the r(UUAAGU) loop has the G8 base protruding into the solvent. The solution structure of the r(UUAAGU) loop is essentially identical to the structure of an identical loop found in the crystal structure of the 30S ribosomal subunit where the guanine in the loop is involved in tertiary interactions with RNA bases from adjacent regions [Wimberly, B. T., Brodersen, D. E., Clemons, W. M., Morgan-Warren, R. J., Carter, A. P., Vonrhein, C., Hartsch, T., and Ramakrishnan, V. (2000) Nature 407, 327-339]. The similarity of the solution and solid-state structures of this hairpin loop suggests that formation of this hairpin may facilitate folding of 16S RNA. PMID- 11502182 TI - Symmetrical stabilization of bound Ca2+ ions in a cooperative pair of EF-hands through hydrogen bonding of coordinating water molecules in calbindin D(9k). AB - Water molecules are found to complete the Ca2+ coordination sphere when a protein fails to provide enough ligating oxygens. Hydrogen bonding of these water molecules to the protein backbone or side chains may contribute favorably to the Ca2+ affinity, as suggested in an earlier study of two calbindin D(9k) mutants [E60D and E60Q; Linse et al. (1994) Biochemistry 33, 12478-12486]. To investigate the generality of this conclusion, another side chain, Gln 22, which hydrogen bonds to a Ca2+-coordinating water molecule in calbindin D(9k), was mutated. Two calbindin D(9k) mutants, (Q22E+P43M) and (Q22N+P43M), were constructed to examine the interaction between Gln 22 and the water molecule in the C-terminal calcium binding site II. Shortening of the side chain, as in (Q22N+P43M), reduces the affinity of binding two calcium ions by a factor of 18 at low ionic strength, whereas introduction of a negative charge, as in (Q22E+P43M), leads to a 12-fold reduction. In 0.15 M KCl, a 7-fold reduction in affinity was observed for both mutants. The cooperativity of Ca2+ binding increases for (Q22E+P43M), while it decreases for (Q22N+P43M). The rates of Ca2+ dissociation are 5.5-fold higher for the double mutants than for P43M at low ionic strength. For both mutants, reduced strength of hydrogen bonding to calcium-coordinating water molecules is a likely explanation for the observed effects on Ca2+ affinity and dissociation. In the apo forms, the (Q22E+P43M) mutant has lower stability toward urea denaturation than (Q22N+P43M) and P43M. 2D (1)H NMR and crystallographic experiments suggest that the structure of (Q22E+P43M) and (Q22N+P43M) is unchanged relative to P43M, except for local perturbations in the loop regions. PMID- 11502183 TI - Kinetic analysis of nonphotochemical quenching of chlorophyll fluorescence. 1. Isolated chloroplasts. AB - Nonphotochemical quenching of chlorophyll fluorescence in plants is indicative of a process that dissipates excess excitation energy from the light-harvesting antenna of photosystem II. The major fraction of quenching is obligatorily dependent upon the thylakoid DeltapH and is regulated by the de-epoxidation state of the xanthophyll cycle carotenoids associated with the light-harvesting complexes. Basic principles of enzyme kinetics have been used to investigate this process in isolated chloroplasts. The extent of quenching was titrated against the estimated thylakoid lumen pH, and a sigmoidal relationship was obtained with a Hill coefficient of 4.5 and a pK of 4.7. Upon de-epoxidation, these parameters changed to 1.6 and 5.7, respectively. Antimycin A suppressed quenching, increasing the Hill coefficient and reducing the pK. The rate of induction of quenching fitted second-order kinetics with respect to illumination time, and the rate constant was dependent upon the DeltapH, the de-epoxidation state, the presence of antimycin, and also the presence of dibucaine, a quenching enhancer. All these data are consistent with the notion that quenching is caused by a conformational transition in a chloroplast thylakoid protein; this transition shows cooperativity with respect to proton binding, and is controlled by de epoxidation state and various exogenous reagents. PMID- 11502184 TI - Kinetic analysis of nonphotochemical quenching of chlorophyll fluorescence. 2. Isolated light-harvesting complexes. AB - The chlorophyll fluorescence yield of purified photosystem II light-harvesting complexes can be lowered by manipulation of experimental conditions. In several important respects, this quenching resembles the nonphotochemical quenching observed in isolated chloroplasts and leaves, therefore providing a model system for investigating the underlying mechanism. A methodology based on the principles of enzyme kinetic analysis has already been applied to isolated chloroplasts, and this same experimental approach was used here with purified LHCIIb, CP26, and CP29. It was found that the kinetics of the decrease in fluorescence yield robustly fitted a second-order kinetic model with respect to time after induction of quenching. The second-order rate constant was dependent upon the complex that was analyzed, the detergent concentration, the solution pH, and the presence of exogenous xanthophyll cycle carotenoids. In contrast, the formation of an absorbance change at 683 nm that accompanies quenching displayed first-order kinetics. The reversal of quenching also displayed second-order kinetics. These data show that quenching results from a binary reaction, possibly arising between two chlorophyll molecules. On the basis of these data, a model for the regulation of nonphotochemical quenching based upon the allosteric control of the conformation of light-harvesting complexes by protonation and xanthophyll binding is presented. PMID- 11502185 TI - N-hydroxyguanidines as new heme ligands: UV-visible, EPR, and resonance Raman studies of the interaction of various compounds bearing a C=NOH function with microperoxidase-8. AB - Interaction between microperoxidase-8 (MP8), a water-soluble hemeprotein model, and a wide range of N-aryl and N-alkyl N'-hydroxyguanidines and related compounds has been investigated using UV-visible, EPR, and resonance Raman spectroscopies. All the N-hydroxyguanidines studied bind to the ferric form of MP8 with formation of stable low-spin iron(III) complexes characterized by absorption maxima at 405, 535, and 560 nm. The complex obtained with N-(4-methoxyphenyl) N' hydroxyguanidine exhibits EPR g-values at 2.55, 2.26, and 1.86. The resonance Raman (RR) spectrum of this complex is also in agreement with an hexacoordinated low-spin iron(III) structure. The dissociation constants (K(s)) of the MP8 complexes with mono- and disubstituted N-hydroxyguanidines vary between 15 and 160 microM at pH 7.4. Amidoximes also form low-spin iron(III) complexes of MP8, although with much larger dissociation constants. Under the same conditions, ketoximes, aldoximes, methoxyguanidines, and guanidines completely fail to form such complexes with MP8. The K(s) values of the MP8-N-hydroxyguanidine complexes decrease as the pH of the solution is increased, and the affinity of the N hydroxyguanidines toward MP8 increases with the pK(a) of these ligands. Altogether these results show that compounds involving a -C(NHR)=NOH moiety act as good ligands of MP8-Fe(III) with an affinity that depends on the electron richness of this moiety. The analysis of the EPR spectrum of the MP8-N hydroxyguanidine complexes according to Taylor's equations shows a strong axial distortion of the iron, typical of those observed for hexacoordinated heme Fe(III) complexes with at least one pi donor axial ligand (HO(-), RO(-), or RS( )). These data strongly suggest that N-hydroxyguanidines bind to MP8 iron via their oxygen atom after deprotonation or weakening of their O-H bond. It thus seems that N-hydroxyguanidines could constitute a new class of strong ligands for hemeproteins and iron(III)-porphyrins. PMID- 11502186 TI - Mutagenic and chemical modification of the ABA-1 allergen of the nematode Ascaris: consequences for structure and lipid binding properties. AB - The polyprotein allergens/antigens of nematodes (NPAs) are the only lipid binding proteins known to be produced as polyproteins. Cleavage of the large polyprotein precursors at regularly spaced proteinase cleavage sites produces 10 or 11 individual protein units of approximately 15 kDa. The sequences of these units are highly diverse within and between species, but there are five absolutely or strongly conserved amino acid positions (Trp15, Gln20, Leu42, Cys64, and Cys120). We have tested the role of these signature amino acids by mutational or chemical alteration of the ABA-1 protein of Ascaris, and examined the resulting modified proteins for perturbations of their lipid binding activities and structural integrity. Substitution of Trp15 and Gln20 both affect the stability of the protein in terms of resistance to thermal or chemical denaturation, but the ligand binding function is unaffected. Mutation of Leu42, however, disrupts both the protein's structural stability and functional integrity, as does chemical disruption of the disulfide bridge formed between Cys64 and Cys120. We also find that the C-terminal, but not the N-terminal, half of the protein binds fatty acids, indicating that the binding site may be confined to this part of the protein. This also supports the idea that the NPA units are themselves derived from an ancient duplication event, and that they may comprise two functionally distinct domains. PMID- 11502187 TI - Membrane binding and self-association of alpha-synucleins. AB - Although its function is unknown, alpha-synuclein is widely distributed in neural tissue and is the major component in the pathological aggregates found in patients with Parkinson's disease, Alzheimer's disease, Down's syndrome, and multiple system atrophy. In this report, we have quantified the binding alpha synucleins to lipid membranes. In contrast to previous studies, we find, using real time equilibrium fluorescence methods, that alpha-synuclein binds strongly to large, unilamellar vesicles with either anionic or zwitterionic headgroups. Membrane binding is also strong for beta-synuclein, phosphorylated alpha synuclein, and a synuclein mutant that is associated with familial Parkinson's disease. In solution at less than 400 nM, synuclein has a tendency to undergo concentration-dependent oligomerization as determined by changes in intrinsic fluorescence and fluorescence resonance energy transfer. Above this concentration, the protein begins to aggregate into structures visible by light scattering. Although membrane binding does not affect the secondary structure of alpha-synuclein, it greatly inhibits the ability of this protein to self associate. Taken together, our results indicate that pathological conditions may be associated with a disruption in synuclein-membrane interactions. PMID- 11502188 TI - Role of terminal nonhomologous domains in initiation of human red cell spectrin dimerization. AB - Human erythrocyte spectrin is an antiparallel heterodimer comprised of a 280 kDa alpha subunit and a 246 kDa beta subunit which further associates into tetramers in the red cell membrane cytoskeleton. Lateral association of the flexible rodlike monomers involves a multiple-step process that is initiated by a high affinity association near the actin-binding end of the molecule (dimer nucleation site). In this study, recombinant alpha and beta proteins comprising two or four "spectrin type" motifs with and without adjacent, terminal nonhomologous domains were evaluated for their relative contributions to dimer initiation, and the thermodynamic properties of these heterodimer complexes were measured. Sedimentation equilibrium studies showed that in the absence of the heterologous subunit, individual recombinant proteins formed weak homodimers (K(d) > 0.3 mM). When 2-motif (alpha20-21 and beta1-2) and 4-motif (alpha18-21 and beta1-4) recombinants lacking the terminal nonhomologous domains were paired with the complementary protein, high affinity heterodimers were formed in sedimentation equilibrium analysis. Both the alpha20-21/beta1-2 complex and the alpha20 21EF/betaABD1-2 complex showed stoichiometric binding with similar binding affinities (K(d) approximately 10 nM) using isothermal titration calorimetry. The alpha20-21/beta1-2 complex showed an enthalpy of -10 kcal/mol, while the alpha20 21EF/betaABD1-2 complex showed an enthalpy of -13 kcal/mol. Pull-down assays using alpha spectrin GST fusion proteins showed strong associations between all heterodimer complexes in physiological buffer, but all heterodimer complexes were destabilized by the presence of Triton X-100 and other detergents. Complexes lacking the nonhomologous domains were destabilized to a greater extent than complexes that included the nonhomologous domains. The detergent effect appears to be responsible for the apparent essential role of the nonhomologous domains in prior reports. Taken together, our results indicate that the terminal nonhomologous domains do not contribute to dimer initiation nor are they required for formation of high affinity spectrin heterodimers in physiological buffers. PMID- 11502189 TI - Shape of Ocr, the gene 0.3 protein of bacteriophage T7: modeling based on light scattering experiments. AB - Ocr, the first protein expressed by bacteriophage T7, inhibits type Iota DNA restriction enzymes by preventing them from binding to DNA. This inhibition allows the phage to successfully infect the host. The shape of ocr is modeled on the basis of static and dynamic light scattering measurements. The static light scattering data confirm previous observations that ocr exists in solution as a dimer. The diffusion constant determined by dynamic light scattering indicates a nonspherical shape of the ocr dimer. Hydrodynamic models of ellipsoids are presented, and it is argued that ocr is best described by a prolate ellipsoid with dimensions of 10.4 nm by 2.6 nm. The size and shape predicted by this model are consistent with ocr acting as a mimic of the DNA structure bound by type Iota restriction enzymes. PMID- 11502190 TI - Thermodynamic characterization of ligand-induced conformational changes in UDP-N acetylglucosamine enolpyruvyl transferase. AB - The binding of UDP-N-acetylglucosamine (UDPNAG) to the enzyme UDP-N acetylglucosamine enolpyruvyl transferase (MurA) was studied in the absence and presence of the antibiotic fosfomycin by isothermal titration calorimetry. Fosfomycin binds covalently to MurA in the presence of UDPNAG and also in its absence as demonstrated by MALDI mass spectrometry. The covalent attachment of fosfomycin affects the thermodynamic parameters of UDPNAG binding significantly: In the absence of fosfomycin the binding of UDPNAG is enthalpically driven (DeltaH = -35.5 kJ mol(-1) at 15 degrees C) and opposed by an unfavorable entropy change (DeltaS = -25 J mol(-1) K(-1)). In the presence of covalently attached fosfomycin the binding of UDPNAG is entropically driven (DeltaS = 187 J mol(-1)K( 1) at 15 degrees C) and associated with unfavorable changes in enthalpy (DeltaH = 28.8 kJ mol(-1)). Heat capacities for UDPNAG binding in the absence or presence of fosfomycin were -1.87 and -2.74 kJ mol(-1) K(-1), respectively, indicating that most ( approximately 70%) of the conformational changes take place upon formation of the UDPNAG-MurA binary complex. The major contribution to the heat capacity of ligand binding is thought to be due to changes in the solvent accessible surface area. However, associated conformational changes, if any, also contribute to the experimentally measured magnitude of the heat capacity. The changes in solvent-accessible surface area were calculated from available 3D structures, yielding a DeltaC(p) of -1.3 kJ mol(-1) K(-1); i.e., the experimentally determined heat capacity exceeds the calculated one. This implies that other thermodynamic factors exert a large influence on the heat capacity of protein-ligand interactions. PMID- 11502191 TI - Theoretical study of a landscape of protein folding-unfolding pathways. Folding rates at midtransition. AB - This paper presents a new method for calculating the folding-unfolding rates of globular proteins. The method is based on solution of kinetic equations for a network of folding-unfolding pathways of the proteins. The rates are calculated in the point of thermodynamic equilibrium between the native and completely unfolded states. The method has been applied to all the proteins listed by Jackson [Jackson, S. E. (1998) Folding Des. 3, R81-R91] and some peptides. Although the studied protein chains differ by more than 1 order of magnitude in size and exhibit two- as well as three-state kinetics in water, and their folding rates cover more than 11 orders of magnitude, the theoretical estimates are reasonable close to the experimentally measured folding rates in midtransition (the correlation coefficient being as high as 0.78). This means that the presented theory (having no adjustable parameters at all) is consistent with the experimental observations. PMID- 11502192 TI - The reaction of serpins with proteinases involves important enthalpy changes. AB - When active serpins are proteolytically inactivated in a substrate-like reaction, they undergo an important structural transition with a resultant increase in their conformational stability. We have used microcalorimetry to show that this conformational alteration is accompanied by an important enthalpy change. For instance, the cleavage of alpha(1)-proteinase inhibitor by Pseudomonas aeruginosa elastase, Staphylococcus aureus V8 proteinase, or papain and that of antithrombin by leukocyte elastase are characterized by large enthalpy changes (DeltaH = -53 to -63 kcal mol(-1)). The former reaction also has a large and negative heat capacity (DeltaC(p)() = -566 cal K(-1) mol(-1)). In contrast, serpins release significantly less heat when they act as proteinase inhibitors. For example, the inhibition of pancreatic elastase, leukocyte elastase, and pancreatic chymotrypsin by alpha(1)-proteinase inhibitor and that of pancreatic trypsin and coagulation factor Xa by antithrombin are accompanied by a DeltaH of -20 to -31 kcal mol(-1). We observe no heat release upon proteolytic cleavage of inactive serpins or following inhibition of serine proteinases by canonical inhibitors or upon acylation of chymotrypsin by N-trans-cinnamoylimidazole. We suggest that part of the large enthalpy change that occurs during the structural transition of serpins is used to stabilize the proteinase in its inactive state. PMID- 11502193 TI - Identification of a region involved in the communication between the NADP(H) binding domain and the membrane domain in proton pumping E. coli transhydrogenase. AB - The two hydrophilic domains I and III of Escherichia coli transhydrogenase containing the binding sites for NAD(H) and NADP(H), respectively, are located on the cytosolic side of the membrane, whereas the hydrophobic domain II is composed of 13 transmembrane alpha-helices, and is responsible for proton transport. In the present investigation the segment betaC260-betaS266 connecting domain II and III was characterized primarily because of its assumed role in the bioenergetic coupling of the transhydrogenase reaction. Each residue of this segment was replaced by a cysteine in a cysteine-free background, and the mutated proteins analyzed. Except for betaS266C, binding studies of the fluorescent maleimide derivative MIANS to each cysteine in the betaC260-betaR266 region revealed an increased accessibility in the presence of NADP(H) bound to domain III; an opposite effect was observed for betaS266. A betaD213-betaR265 double cysteine mutant was isolated in a predominantly oxidized form, suggesting that the corresponding residues in the wild-type enzyme are closely located and form a salt bridge. The betaS260C, betaK261C, betaA262C, betaM263, and betaN264 mutants showed a pronounced inhibition of proton-coupled reactions. Likewise, several betaR265 mutants and the D213C mutant showed inhibited proton-coupled reactions but also markedly increased values. It is concluded that the mobile hinge region betaC260-betaS266 and the betaD213-betaR265 salt bridge play a crucial role in the communication between the proton translocation/binding events in domain II and binding/release of NADP(H) in domain III. PMID- 11502194 TI - Specific binding of 8-oxoguanine-containing RNA to polynucleotide phosphorylase protein. AB - 8-Oxoguanine, an oxidized form of guanine, has the potential to pair with both cytosine and adenine, and thus, the persistence of this base in messenger RNA would cause translational errors. To prevent such an outcome, organisms probably have a mechanism for recognizing RNA molecules carrying 8-oxoguanine and prevent them from entering into the cellular translational machinery. We now report that the Escherichia coli cell possesses proteins that bind specifically to RNA carrying 8-oxoguanine. On incubation with a cell-free extract, 8-oxoguanine containing RNA is stable while normal RNA is degraded by cellular nucleases. The RNase protection assay and gel shift assay revealed that some proteins bind specifically to 8-oxoguanine-containing RNA, hence preventing nuclease attacks. Among the complexes that were detected, one with a 77 kDa protein exhibits tight binding between RNA and protein components. This protein was identified as polynucleotide phosphorylase, encoded by the pnp gene. pnp(-)() mutants are hyperresistant to paraquat, a drug that induces oxidative stress in the cell. Binding of Pnp protein to 8-oxoguanine-containing RNA would inhibit cell growth, probably due to withdrawal of such RNA from the translational machinery. The Pnp protein may, therefore, discriminate between an oxidized RNA molecule and a normal one, thus contributing a high fidelity of translation. PMID- 11502195 TI - Conformation of the C-terminal domain of the pro-apoptotic protein Bax and mutants and its interaction with membranes. AB - The C-terminal domain of the pro-apoptotic protein Bax is a hydrophobic stretch which, it has been predicted, anchors this protein to the outer mitochondrial membrane when apoptosis is induced in the cell. A 21mer peptide imitating this domain has been synthesized together with two mutants, one with a S184 substituted by K and the other with the S184 deleted. When their structures were studied by infrared spectroscopy, it was seen that the three peptides formed aggregates both in solution and within lipid membranes, and that the peptide changed its secondary structure as a consequence of these two mutations. It was also observed that the wild-type peptide and the two mutants became membrane integral molecules and changed their conformation when they were incorporated into model membranes with the same composition as the outer mitochondrial membrane. With the peptides incorporated in the membranes the location of W188 was studied by fluorescence quenching using the water soluble quencher acrylamide and different doxyl-PC located in the membrane, this residue being found at different membrane depths in each of the three peptides. The fact that the three peptides were able to perturb the motion of the fluorescent probe diphenylhexatriene confirmed their insertion in the membrane. However, whereas the wild type and the DeltaS184 mutant peptides were very efficient in releasing encapsulated carboxyfluorescein from liposomes, the mutant S184K was less efficient. Taken together, these results showed that the mutation tested changed the conformation of the C-terminal domain of Bax and the positions that they adopted when inserted in membranes, confirming the importance of S184 determining the conformation of this domain. At the same time, these results confirmed that the C-terminal domain of Bax participates in disrupting the barrier properties of biomembranes. PMID- 11502197 TI - Psychrophilic valine dehydrogenase of the antarctic psychrophile, Cytophaga sp. KUC-1: purification, molecular characterization and expression. AB - We found the occurrence of valine dehydrogenase in the cell extract of a psychrophilic bacterium, Cytophaga sp. KUC-1, isolated from Antarctic seawater and purified the enzyme to homogeneity. The molecular mass of the enzyme was determined to be approximately 154 kDa by gel filtration and that of the subunit was 43 kDa by SDS/PAGE: the enzyme was a homotetramer. The enzyme required NAD+ as a coenzyme, and catalyzed the oxidative deamination of L-valine, L-isoleucine, L-leucine and the reductive amination of alpha-ketoisovalerate, alpha ketovalerate, alpha-ketoisocaproate, and alpha-ketocaproate. The reaction proceeds through an iso-ordered bi-bi mechanism. The enzyme was highly susceptible to heat treatment and the half-life at 45 degrees C was estimated to be 2.4 min. The kcat/Km (micro(-1).s(-1)) values for L-valine and NAD+ at 20 degrees C were 27.48 and 421.6, respectively. The enzyme showed pro-S stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of coenzyme. The gene encoding valine dehydrogenase was cloned into Escherichia coli (Novablue), and the primary structure of the enzyme was deduced on the basis of the nucleotide sequence of the gene encoding the enzyme. The enzyme contains 370 amino-acid residues, and is highly homologous with S. coelicolor ValDH (identity, 46.7%) and S. fradiae ValDH (43.1%). Cytophaga sp. KUC-1 ValDH contains much lower numbers of proline and arginine residues than those of other ValDHs. The changes probably lead to an increase in conformational flexibility of the Cytophaga enzyme molecule to enhance the catalytic activity at low temperatures. PMID- 11502196 TI - Concerted influence of key amino acids on the lipid binding properties of a single-spanning membrane protein: NMR and mutational analysis. AB - Finding the combinations of key amino acids involved in the interaction network underlying the interfacial features of membrane proteins would contribute to a better understanding of their sequence-structure-function relationships and the role of anionic phospholipids. To further address these questions, we performed mutational analysis associated with NMR experiments on synthetic fragments of the single-spanning membrane protein PMP1 that exhibit binding specificity for phosphatidylserine (PS). The aromatic and glutamine residues of the helix part of the PMP1 cytoplasmic domain were mutated. (1)H NMR experiments were carried out using perdeuterated DPC micelles as a membrane-like environment, in the absence and presence of small amounts of either POPC or POPS lipids. From intermolecular NOEs and chemical shift data, specific and nonspecific aspects of peptide phospholipid interactions were distinguished. The major finding of our study is to reveal the concerted influence of a tryptophan and a glutamine residue on the interfacial conformation and lipid binding specificity of the PMP1 cytoplasmic domain. PMID- 11502198 TI - The molecular interaction of human salivary histatins with polyphenolic compounds. AB - Dietary tannins are polyphenols that are effectively precipitated by salivary histatins (Hsts), a novel family of tannin binding proteins. Epigallocatechin gallate (EGCG), a flavan-3-ol ester related to condensed tannins (polymerized products of flavan-3-ols), and pentagalloyl glucose (PGG), a hydrolyzable tannin, were used to evaluate the molecular nature of Hst-polyphenol interaction. NMR demonstrated that Hst5, a representative Hst, bound to EGCG in a hydrophobic manner via basic and aromatic residues. In contrast, proline plays a dominant role in polyphenol binding to other tannin precipitating proteins. The role of basic and aromatic amino acids in EGCG binding was investigated using a series of modified Hsts in each of which one type of amino acid was substituted by Ala. EGCG bound to all modified Hsts, but the binding was diminished. Optimal EGCG binding also depended on the primary structure, as a polypeptide with randomised Hst5 sequence showed significantly diminished interaction with EGCG. Soluble EGCG/Hst5 complexes containing up to seven molecules of EGCG per mol of Hst5 had a 1-mM dissociation constant. In contrast to EGCG, PGG formed small soluble complexes with Hst5 consisting of only one molecule each of PGG and Hst5, as demonstrated by analytical ultracentrifugation. These complexes became insoluble upon binding of additional molecules of PGG. Diminished PGG binding was seen to a peptide with a Hst5 randomized sequence showing the importance of the primary structure. Hsts may serve to form insoluble complexes with tannins thereby preventing their absorption from the intestines and potentially harmful biological effects. In contrast the much weaker interaction with EGCG may allow its uptake into the organism and exploitation of its antioxidant effect. PMID- 11502199 TI - Identification and cloning of an 85-kDa protein homologous to RING3 that is upregulated in proliferating endothelial cells. AB - A central event in angiogenesis is proliferation of blood vessels, which plays a major role in the progression of a number of inflammatory and neoplastic diseases. It is responsible for the switch of endothelial cells from an antiangiogenic to an angiogenic phenotype. To identify novel activated/proliferating-related proteins in human endothelial cells, a subtractive immunization approach was used to elicit a host antibody response against human dermal microvascular endothelial cells (HDMECs) stimulated with a potent angiogenic cytokine such as VPF/VEGF165. In this study, a new mAb, LY9, which is highly specific to VPF/VEGF165-activated HDMECs, was isolated. Stimulation of HDMECs by VPF/VEGF165 or basic fibroblast growth factor (bFGF) resulted in a dose-dependent and time-dependent increase in the binding of LY9. On Western-blot analysis, LY9 identified an 85-kDa protein (p85) in the lysates of several endothelial cells derived from microvascular or large vessel sources, the expression of which is dramatically increased by VPF/VEGF165. The mAb also identified p85 in primary cell cultures of human foreskin keratinocytes but failed to recognize human fibroblasts (MRC5) and a number of different human tumor cell lines, including MG63 osteosarcoma and MCF7 breast carcinoma cells. Immunological screening of a human keratinocyte lambdagt11 cDNA expression library with LY9 identified a partial cDNA clone of 750 bp. DNA sequencing of this clone and predicted amino acids showed more than 93% homology to RING3 kinase, a member of a newly described family of bromodomain-containing proteins that transactivates in the nucleus the promoters of a number of the E2F family of transcription factors. This molecule may represent a new signaling target activated by VPF/VEGF165 and bFGF that allows endothelial cells to enter the proliferative phase of the angiogenic process. PMID- 11502200 TI - Characterization of PSA-RP2, a protein related to prostate-specific antigen and encoded by alternative hKLK3 transcripts. AB - Despite the wide use of prostate-specific antigen (PSA) as a marker of prostate cancer, analysis of its gene products has not yet been completed. The structure of two alternative mRNAs (0.9 and 1.65 kb) of the hKLK3 gene that retain the third intron is reported here. These partially spliced transcripts were detected by hybridization or RT-PCR in normal prostate tissue, benign prostate hyperplasia (BPH) and cancerous prostate tissues, and also in the prostate LNCaP cell line. Insertion of the unspliced intron creates an in-frame stop codon and results in a truncated prepro PSA variant of 180 amino-acid residues. This novel variant, designated PSA-RP2, has an alternate C-terminal tail and lacks the serine residue essential for the catalytic activity of PSA. Prepro PSA-RP2 was transiently produced in COS-7 cells and detected in the spent medium using an anti-PSA serum. Secreted PSA-RP2 was glycosylated with an apparent molecular mass of 25 kDa. Our findings suggest that PSA-RP2 contributes to the molecular heterogeneity of free PSA in the serum of patients with benign or malignant prostate tumors. PMID- 11502201 TI - The amino-acid sequence of the glucose/mannose-specific lectin isolated from Parkia platycephala seeds reveals three tandemly arranged jacalin-related domains. AB - A mannose/glucose-specific lectin was isolated from seeds of Parkia platycephala, the most primitive subfamily of Leguminosae plants. The molecular mass of the purified lectin determined by mass spectrometry was 47 946 +/- 6 Da (by electrospray ionization) and 47 951 +/- 9 Da (by matrix-assisted laser-desoption ionization). The apparent molecular mass of the lectin in solutions of pH in the range 4.5-8.5 determined by analytical ultracentrifugation equilibrium sedimentation was 94 +/- 3 kDa, showing that the protein behaved as a non-pH dependent dimer. The amino-acid sequence of the Parkia lectin was determined by Edman degradation of overlapping peptides. This is the first report of the primary structure of a Mimosoideae lectin. The protein contained a blocked N terminus and a single, nonglycosylated polypeptide chain composed of three tandemly arranged homologous domains. Each of these domains shares sequence similarity with jacalin-related lectin monomers from Asteraceae, Convolvulaceae, Moraceae, Musaceae, Gramineae, and Fagaceae plant families. Based on this homology, we predict that each Parkia lectin repeat may display a beta prism fold similar to that observed in the crystal structure of the lectin from Helianthus tuberosus. The P. platycephala lectin also shows sequence similarity with stress- and pathogen-upregulated defence genes of a number of different plants, suggesting a common ancestry for jacalin-related lectins and inducible defence proteins. PMID- 11502202 TI - Hepatocyte growth factor/scatter factor stimulates migration of rat mammary fibroblasts through both mitogen-activated protein kinase and phosphatidylinositol 3-kinase/Akt pathways. AB - Hepatocyte growth factor/scatter factor (HGF/SF) is considered to be a mesenchymal-derived factor that acts via a dual system receptor, consisting of the MET receptor and proteoglycans present on adjacent epithelial cells. Surprisingly, HGS/SF stimulated the migration of rat mammary (Rama) 27 fibroblasts, although it failed to stimulate their proliferation. HGF/SF stimulated a transient activation of mitogen-activated protein kinases p44 and p42 (p42/44(MAPK)), with a maximum level of dual phosphorylation of p42/44(MAPK) occurring 10-15 min after the addition of the growth factor, which was followed by a rapid decrease to near basal levels after 20 min. Interestingly, a second phase of p42/44(MAPK) dual phosphorylation was observed at later times (3 h to 10 h). PD098059, a specific inhibitor of MEK-1, prevented the dual phosphorylation of p42/44(MAPK) and also the phosphorylation of p90(RSK) (ribosomal subunit S6 kinase), which mirrored the kinetics of p42/44(MAPK) phosphorylation. Moreover, PD098059 prevented the HGF/SF-induced migration of Rama 27 cells. HGF/SF also induced an early increase in the phosphorylation of protein kinase B/Akt. Akt phosphorylation was elevated 15 min after the addition of HGF/SF and then declined to basal levels by 30 min. Wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PtdIns3K), prevented the increase in Akt phosphorylation and abolished HGF/SF-induced migration of fibroblasts. PD098059 also inhibited the stimulation of Akt phosphorylation by HGF/SF and wortmannin similarly inhibited the stimulation of p42/44(MAPK) dual phosphorylation. These results suggest that HGF/SF-induced motility depends on both the transient dual phosphorylation of p42/44(MAPK) and the activation of PtdIns3K in Rama 27 fibroblasts and that these pathways are mutually dependent. PMID- 11502203 TI - Localization of epitopes for monoclonal antibodies to urokinase-type plasminogen activator: relationship between epitope localization and effects of antibodies on molecular interactions of the enzyme. AB - We localized the epitopes for several murine mAbs to human urokinase-type plasminogen activator (uPA) by Ala scanning mutagenesis and related the localization to the effects of the mAbs on the molecular interactions of uPA. Several antibodies against the serine proteinase domain (SPD) were found to have overlapping epitopes composed of variable combinations of Arg178, Arg179, His180, Arg181, Tyr209, Lys211, and Asp214 in the so-called 37-loop and 60-loop, located near the active site and taking part in the binding of uPA to plasminogen activator inhibitor-1 (PAI-1). Besides inhibiting uPA-catalysed plasminogen activation, all antibodies to SPD strongly delayed the binding of uPA to PAI-1, decreasing the second-order rate constant 15- to 6500-fold. There was no correlation between the relative effects of the 37-loop and 60-loop substitutions on the second-order rate constant and on the binding of the antibodies, indicating that the antibodies did not delay complex formation by blocking residues of specific importance for the uPA-PAI-1 reaction, but rather by steric hindrance of the access of PAI-1 to the active site. The affinity of the SPD antibodies for the uPA-PAI-1 complex was only slightly lower than that for free uPA, indicating that the 37-loop and 60-loop are exposed in the complex. The epitopes for two antibodies to the kringle included Arg108, Arg109, and Arg110. The ability of these antibodies to block the binding of uPA to polyanions correlated with a reduced uPA-polyanion affinity after substitution of the three Arg residues. PMID- 11502204 TI - Identification of rat cyclic nucleotide phosphodiesterase 11A (PDE11A): comparison of rat and human PDE11A splicing variants. AB - We have isolated and characterized rat cyclic nucleotide phosphodiesterase (PDE)11A, which exhibits properties of a dual-substrate PDE, and its splice variants (RNPDE11A2, RNPDE11A3, and RNPDE11A4). The deduced amino-acid sequence of the longest form of rat PDE11A splice variant, RNPDE11A4, was 94% identical with that of the human variant (HSPDE11A4). Rat PDE11A splice variants were expressed in a tissue-specific manner. RNPDE11A4 showed unique tissue distribution distinct from HSPDE11A4, which is specifically expressed in the prostate. Rat PDE11A splice variants were expressed in COS-7 cells, and their enzymatic characteristics were compared. Although the Km values for cAMP and cGMP were similar for all of them (1.3-1.6 and 2.1-3.9 microM, respectively), the Vmax values differed significantly (RNPDE11A4 >> RNPDE11A2 > RNPDE11A3). Human PDE11A variants also displayed very similar Km values and significantly different Vmax values (HSPDE11A4 >> HSPDE11A2 > HSPDE11A3 >> HSPDE11A1). The Vmax values of HSPDE11A4 for cAMP and cGMP were at least 100 times higher than those of HSPDE11A1. These observations indicate unique characteristics of PDE11A splicing variants. PMID- 11502205 TI - Internalization of tenecin 3 by a fungal cellular process is essential for its fungicidal effect on Candida albicans. AB - Tenecin 3 is a glycine-rich, antifungal protein of 78 residues isolated from the insect Tenebrio molitor larva. As an initial step towards understanding the antifungal mechanism of tenecin 3, we examined how this protein interacts with the pathogenic fungus Candida albicans to exert its antifungal action. Tenecin 3 did not induce the release of a fluorescent dye trapped in the artificial membrane vesicles and it did not perturb the membrane potential of C. albicans by the initial interaction. Fluorescence confocal microscopy and flow cytometric analysis revealed that tenecin 3 is rapidly internalized into the cytoplasmic space in energy-dependent and temperature-dependent manners. This internalization is also dependent on the ionic environment and cellular metabolic states. These results suggest that the internalization of tenecin 3 into the cytoplasm of C. albicans is mediated by a fungal cellular process. The internalized tenecin 3 is dispersed in the cytoplasm, and the loss of cell viability occurs after this internalization. PMID- 11502206 TI - Drug sequestration in cytoplasmic organelles does not contribute to the diminished sensitivity of anthracyclines in multidrug resistant K562 cells. AB - Cells that acquire multidrug resistance (MDR) are characterized by a decreased accumulation of a variety of drugs. In addition, sequestration of drugs in intracellular vesicles has often been associated with MDR. However, the nature and role of intracellular vesicles in MDR are unclear. We addressed the relationship between MDR and vesicular anthracycline accumulation in the erythroleukemia cell line K562 and a drug-resistant counterpart K562/ADR that overexpresses P-glycoprotein. We used four anthracyclines (all of which are P glycoprotein substrates): daunorubicin and idarubicin, which have good affinity for DNA and as weak bases can accumulate inside acidic compartments; hydroxyrubicin, which binds to DNA but is uncharged at physiological or acidic pH and thus cannot accumulate in acidic compartments; and WP900, an enantiomer of daunorubicin, which is a weak DNA binder but has the same pKa and lipophilicity as daunorubicin. The intrinsic fluorescence of anthracyclines allowed us to use macro- and micro-spectrofluorescence, flow cytometry, and confocal microscopy to characterize their nuclear or intravesicular accumulation in living cells. We found that vesicular accumulation of daunorubicin, WP900 and idarubicin, containing a basic 3'-amine was predominantly restricted to lysosomes in both cell lines, that pH regulation of acidic compartments was not defective in human K562 cells, and that vesicular drug accumulation was much more pronounced in the parental tumor cell line than in the multidrug-resistant cells. These results indicate that vesicular anthracycline sequestration does not contribute to the diminished sensitivity to anthracyclines in multidrug-resistant K562 cells. PMID- 11502207 TI - A further clue to understanding the mobility of mitochondrial yeast cytochrome c: a (15)N T1rho investigation of the oxidized and reduced species. AB - A new approach was developed to overproduce 15N-enriched yeast iso-1-cytochrome c in the periplasm of Escherichia coli in order to perform a study of the motions in the ms-micros time scale on the oxidized and reduced forms through rotating frame 15N relaxation rates and proton/deuterium exchange studies. It is confirmed that the reduced protein is rather rigid whereas the oxidized species is more flexible. The regions of the protein that display increased internal mobility upon oxidation are easily identified by the number of residues experiencing conformational equilibria and by their exchange rates. These data complement the information already available in the literature and provide a comprehensive picture of the mobility in the protein. In particular, oxidation mobilizes the loop containing Met80 and, through specific contacts, affects the mobility of helix 3 and possibly of helix 5, and of a section of protein connecting the heme propionates to helix 2. The relevance of internal motions to molecular recognition and to the early steps of the unfolding process of the oxidized species is also discussed. In agreement with the reported data, subnanosecond mobility is found to be less informative than the ms-micros with respect to redox dependent properties. PMID- 11502208 TI - Dimerization, stability and electrostatic properties of porcine beta lactoglobulin. AB - The study of homologous proteins belonging to the same family can provide a rationale for important molecular properties such as oligomer formation, folding mechanism and mode of binding. We report here a physico-chemical characterization of porcine beta-lactoglobulin, purified from pooled milk: size-exclusion chromatography, CD and NMR measurements were used to study the aggregation and stability of this protein. In spite of the high sequence identity and homology of porcine beta-lactoglobulin with the widely studied bovine species, the two proteins exhibit very different behaviours. The porcine protein shows a monomer dimer equilibrium with a pH dependence opposite to that observed for the bovine species. Unfolding experiments revealed the presence of an intermediate that probably has excess alpha helices, as reported for equine species. Modelling studies were performed on bovine, porcine and equine proteins, and, interestingly, electrostatic surface potential calculations led to results consistent with the different dimer interface found for porcine beta lactoglobulin in the crystal structure. Interaction studies revealed that porcine beta-lactoglobulin is unable to bind fatty acids at any pH, thus questioning the main functional role proposed for lactoglobulins as fatty acid transporters or solubilizers. PMID- 11502209 TI - Oxidized low density lipoproteins downregulate LPS-induced platelet-activating factor receptor expression in human monocyte-derived macrophages: implications for LPS-induced nuclear factor-kappaB binding activity. AB - Monocytes/macrophages play a key role in atherogenesis due to their inflammatory properties including formation of lipid mediators such as platelet-activating factor (PAF). We investigated the effect of oxidized low-density lipoprotein (oxLDL) on lipopolysaccharide (LPS)-induced PAF receptor (PAF-R) expression in human macrophages and the implication of the nuclear factor (NF)-kappaB in this regulation. LPS-treatment (1 microg.mL(-1)) of macrophages increased PAF binding and PAF-R mRNA expression by 56% (P < 0.05) and twofold (P < 0.01), respectively. In contrast, highly oxidized low-density lipoprotein [ox24hLDL; 100 microg.mL( 1); thiobarbituric acid reacting substances: 31 +/- 3 nmol equiv. malondialdehyde (MDA).mg protein LDL-1] diminished PAF-R expression (-69%; P < 0.05) and mRNA level (- 45%; P < 0.01). LPS pretreatment induced the activated form of p65 in the nuclear compartment of macrophages (detected by Western blotting) and NF kappaB binding activity (by electrophoretic mobility shift assay). Treatment of macrophages with ox24hLDL suppressed the LPS-induced binding of NF-kappaB to DNA. In addition, treatment of macrophages with lysophosphatidylcholine (2 and 10 microM), a major component of oxLDL, inhibited the LPS-induced NF-kappaB binding to DNA and reduced PAF binding by 30 and 70%, respectively. In conclusion, oxLDL may downregulate PAF-R expression in human macrophages by inhibiting LPS-induced NF-kappaB binding to DNA. PMID- 11502210 TI - Intrinsic versus extrinsic stabilization of enzymes: the interaction of solutes and temperature on A4-lactate dehydrogenase orthologs from warm-adapted and cold adapted marine fishes. AB - We examined the effects of temperature and stabilizing solutes on A4-lactate dehydrogenase (A4-LDH) from warm- and cold-adapted fishes, to determine how extrinsic stabilizers affect orthologs with different intrinsic stabilities. Conformational changes during substrate binding are rate-limiting for A4-LDH, thus stabilization due to intrinsic or extrinsic factors leads to decreased activity. A4-LDH from a warm-temperate goby (Gillichthys mirabilis), which has lower values for kcat and the Michaelis constant for pyruvate ( K m PYR), was intrinsically more stable than the orthologs of the cold-adapted Antarctic notothenioids Parachaenichthys charcoti and Chionodraco rastrospinosus, as shown by a higher apparent transition ('melting') temperature (Tm(APP)). We used four solutes, glycerol, sucrose, trimethylamine-N-oxide and poly(ethylene glycol) 8000, which stabilize proteins through different modes of preferential exclusion, to study temperature-solute interactions of the three orthologs. Changes in Tm(APP) were similar for all orthologs in each solute tested, but the catalytic rate of G. mirabilis A4-LDH was decreased most by solutes and increased most by temperature. In contrast, the K m PYR values of the Antarctic orthologs were more affected than that of the goby by both solutes and temperature. We conclude that (a) preferential exclusion of solutes functions within the native state of A4-LDH to favor conformational microstates with minimal surface area; (b) the varied effects of the different solutes on the kinetic properties are due to the interaction between this nonspecific stabilization and the differing intrinsic stabilities of the orthologs; (c) the catalytic rates of A4-LDH orthologs are equally affected by stabilizing solutes, if measurements are made at physiologically appropriate temperatures; and (d) global stability and localized flexibility of these A4-LDH orthologs may evolve independently. PMID- 11502211 TI - Expression of the rabbit cytochrome P450 aromatase encoding gene uses alternative tissue-specific promoters. AB - The aim of the present study was to analyse the tissue-specific expression of various promoter-derived transcripts from the gene encoding rabbit aromatase cytochrome P450. A new promoter, named I.r, was identified, and promoters II and I.r were sequenced. Promoter I.r-derived transcripts were found in preovulatory granulosa cells, corpus luteum, placenta and adipose tissue. An alternative splice variant of this transcript was found with tissue-specific preference. Tissue-specific expression of promoter-derived variants was studied in the ovary before and after ovulation. While the level of promoter II-derived transcript decreased dramatically after ovulation, that of promoter I.r-derived transcript remained unchanged, indicating that promoter II and promoter I.r were not controlled by a single regulation system. The existence of this dual system of regulation suggests that the rabbit ovary could be a useful model to study the promoter-specific regulation of aromatase. PMID- 11502212 TI - Focal adhesion kinase affects the sensitivity of human hepatocellular carcinoma cell line SMMC-7721 to tumor necrosis factor-alpha/cycloheximide-induced apoptosis by regulating protein kinase B levels. AB - Most cell lines are resistant to tumor necrosis factor-alpha (TNF-alpha) cytotoxicity and require cotreatment of TNF-alpha with cycloheximide (Chx) to undergo apoptosis. Recently, the serine/threonine protein kinase, protein kinase B has been demonstrated to protect cells from apoptosis induced by TNF-alpha. In this study, we have shown that the human hepatocellular carcinoma cell line, SMMC 7721, was insensitive to TNF-alpha cytotoxicity and underwent apoptosis quickly in the presence of TNF-alpha and Chx. PKB levels decreased during TNF-alpha/Chx induced apoptosis. No significant change in PKB levels was found in the presence of TNF-alpha or Chx alone. It seemed that the level of PKB closely correlated with apoptosis. The protein level of focal adhesion kinase (FAK) was reduced by 66% by transfecting FAK antisense cDNA recombinant vector into SMMC-7721 cells. We determined the apoptosis-induced effect of TNF-alpha/Chx on the FAK antisense cDNA transfectant cells. The results indicated that the percentage of apoptotic cells was enhanced at lower doses of TNF-alpha (10, 20 or 50 U.mL(-1)) and decreased at a higher dose of TNF-alpha (1000 U.mL(-1)) in the transfected cells as compared to the control. Correspondingly, in the FAK antisense cDNA transfectant cells treated with lower doses of TNF-alpha in presence of 10 microg.mL(-1) Chx, the PKB level was lower, but in the FAK antisense cDNA transfectants treated with higher doses of TNF-alpha in presence of 10 microg.mL( 1) Chx, the PKB level was higher. In response to TNF-alpha alone, FAK antisense cDNA transfectants showed a decrease in the level of PKB. However, in the case of TNF-alpha cotreated with wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PtdIns3K), the FAK antisense cDNA transfectants produced significantly less amounts of PKB than the control. It seemed that FAK could stimulate PKB levels through a pathway not involving PtdIns3K. These results suggest that FAK can affect the sensitivity of SMMC-7721 cells to TNF-alpha/Chx-induced apoptosis in a biphasic manner by regulating PKB levels. PMID- 11502213 TI - Mechanism of substrate inhibition in cellulose synergistic degradation. AB - A comprehensive experimental study of substrate inhibition in cellulose hydrolysis based on a well defined system is presented. The hydrolysis of bacterial cellulose by synergistically operating binary mixtures of cellobiohydrolase I from Trichoderma reesei and five different endoglucanases as well as their catalytic domains displays a characteristic substrate inhibition. This inhibition phenomenon is shown to require the two-domain structure of an intact cellobiohydrolase. The experimental data were in accordance with a mechanism where cellobiohydrolases previously bound to the cellulose by means of their cellulose binding domains are able to find chain ends by lateral diffusion. An increased substrate concentration at a fixed enzyme load will also increase the average diffusion distance/time needed for cellobiohydrolases to reach new chain ends created by endoglucanases, resulting in an apparent substrate inhibition of the synergistic action. The connection between the binding properties and the substrate inhibition is encouraging with respect to molecular engineering of the binding domain for optimal performance in biotechnological processes. PMID- 11502214 TI - A mouse in vitro transcription system reconstituted from highly purified RNA polymerase II, TFIIH and recombinant TBP, TFIIB, TFIIE and TFIIF. AB - Unregulated transcription of protein-encoding genes in vitro is dependent on 12 subunit core RNA polymerase II and five general transcription factors; TATA binding protein (TBP), transcription factor (TF)IIB, TFIIE, TFIIF, and TFIIH. Here we describe cloning of the mouse cDNAs encoding TFIIB and the small and large TFIIE and TFIIF subunits. The cDNAs have been used to express the corresponding proteins in recombinant form in Escherichia coli and in Sf21 insect cells, and all proteins have been purified to > 90% homogeneity. We have also purified a recombinant His6-tagged mouse TBP to near homogeneity and show that it is active in both a reconstituted mouse in vitro transcription system and a TBP dependent in vitro transcription system from Saccharomyces cerevisiae. The more complex general transcription factors, TFIIH and RNA polymerase II, were purified more than 1000-fold and to near homogeneity, respectively, from tissue cultured mouse cells. When combined, the purified factors were sufficient to initiate transcription from different promoters in vitro. Functional studies of the S phase-specific mouse ribonucleotide reductase R2 promoter using both the highly purified system described here (a mouse cell nuclear extract in vitro transcription system) and in vivo R2-promoter reporter gene assays together identify an NF-Y interacting promoter proximal CCAAT-box as being essential for high-level expression from the R2 promoter. PMID- 11502215 TI - Cytochrome c reconstituted from two peptide fragments displays native-like redox properties. AB - Recombination of two fragments of horse cytochrome c (the heme-containing N fragment, residues 1-56, and the C-fragment, residues 57-104), which are substantially unstructured at neutral pH, gives rise to a 1:1 fragment complex with a compact conformation, in which the alpha helical structure and the native Met80-Fe(III) axial bond are recovered. With respect to the native protein, the ferric complex shows a less rigid atomic packing and a decreased stability [Delta(DeltaG(o))D = 14.7 kJ.mol(-1)], ascribed to perturbations involving the Trp59 microenvironment and, to a lower extent, the heme pocket region. The redox potential, E1/2 = 234 +/- 5 mV vs. normal hydrogen electrode at 25 degrees C, is close to that of the intact protein, consistent with recovery of the native Met80 heme Fe(III) axial bond. Furthermore, the fragment complex shows reactivity similar to intact cytochrome c, in the reaction with cytochrome c oxidase. We conclude that the absence in the complex of some native cross-links and interlocked packing important for protein rigidity and stability is not as relevant for maintaining the native redox properties of the protein, provided that some structural requirements (i.e. recovering of the native-like alpha helical structure) are fulfilled and coordination of Met80 to the heme-iron is restored. PMID- 11502216 TI - Assembly of gap junction channels: mechanism, effects of calmodulin antagonists and identification of connexin oligomerization determinants. AB - The assembly of connexins (Cxs) into gap junction intercellular communication channels was studied. An in vitro cell-free synthesis system showed that formation of the hexameric connexon hemichannels involved dimeric and tetrameric connexin intermediates. Cx32 contains two putative cytoplasmic calmodulin-binding sites, and their role in gap junction channel assembly was investigated. The oligomerization of Cx32 into connexons was reversibly inhibited by a calmodulin binding synthetic peptide, and by W7, a naphthalene sulfonamide calmodulin antagonist. Removing the calmodulin-binding site located at the carboxyl tail of Cx32 limited connexon formation and resulted in an accumulation of intermediate connexin oligomers. This truncation mutant, Cx32Delta215, when transiently expressed in COS-7 cells, accumulated intracellularly and had failed to target to gap junctions. Immunoprecipitation studies suggested that a C-terminal sequence of Cx32 incorporating the calmodulin-binding site was required for the formation of hetero-oligomers of Cx26 and Cx32 but not for Cx32 homomeric association. A chimera, Cx32TM3CFTR, in which the third transmembrane and proposed channel lining sequence of Cx32 was substituted by a transmembrane sequence of the cystic fibrosis transmembrane conductance regulator, did not oligomerize in vitro and it accumulated intracellularly when expressed in COS-7 cells. The results indicate that amino-acid sequences in the third transmembrane domain and a calmodulin binding domain in the cytoplasmic tail of Cx32 are likely candidates for regulating connexin oligomerization. PMID- 11502217 TI - Evidence for efficient uptake and incorporation of sialic acid by eukaryotic cells. AB - Sialic acids are the most abundant terminal carbohydrate moiety on cell surface glycoconjugates in eukaryotic cells and are of functional importance for many biological ligand-receptor interactions. It is a widely accepted view that sialic acids cannot be efficiently taken up from the extracellular space by eukaryotic cells. To test this assumption, we cultivated two recently identified human hematopoetic cell lines which are hyposialylated due to a deficiency in de novo sialic acid biosynthesis in the presence of N-acetylneuraminic acid (NeuAc), the most frequently found sialic acid. Surprisingly, NeuAc medium supplementation rapidly and potently compensated for the endogenous hyposialylation in a concentration-dependent manner, resulting in the presentation of cell surface sialoglycans involved in cell adhesion, virus infection and signal transduction. We provide several lines of experimental evidence that all suggest that NeuAc was neither extracellularly incorporated nor degraded to a less complex sugar before uptake. Importantly, NeuAc induced a marked increase in intracellular CMP-NeuAc levels in both human cell lines and in primary cells regardless of the prior sialylation status of the cells. Studies employing 9-[3H]NeuAc revealed an uptake consistent with the observed incorporation of unlabeled NeuAc. We propose the existence of an efficient uptake mechanism for NeuAc in eukaryotic cells. PMID- 11502218 TI - Rapid, ATP-dependent degradation of a truncated D1 protein in the chloroplast. AB - The D1 protein constitutes one of the reaction center subunits of photosystem II and turns over rapidly due to photooxidative damage. Here, we studied the degradation of a truncated D1 protein. A plasmid with a precise deletion in the reading frame of the psbA gene encoding D1 was introduced into the chloroplast of Chlamydomonas reinhardtii. A homoplasmic mutant containing the desired gene was able to synthesize the truncated form of the polypeptide, but could not accumulate significant levels of it. As a consequence, other central photosystem II subunits did not assemble within the thylakoid membrane. In vivo pulse-chase experiments showed that the abnormal D1 protein is rapidly degraded in the light. Degradation was delayed in the light in the presence of an uncoupler, or when cells were incubated in the dark. Pulse-chase experiments performed in vitro indicate that an ATP and metal-dependent protease is responsible for the breakdown process. The paper describes the first in vivo and in vitro functional test for ATP-dependent degradation of a defect polypeptide in chloroplasts. The possible involvement of proteases similar to those removing abnormal proteins in prokaryotic organisms is discussed on the basis of proteases recently identified in chloroplasts. PMID- 11502219 TI - Secretion of biologically active glycoforms of bovine follicle stimulating hormone in plants. AB - We chose the follicle stimulating hormone (FSH), a pituitary heterodimeric glycoprotein hormone, as a model to assess the ability of the plant cell to express a recombinant protein that requires extensive N-glycosylation for subunit folding and assembly, intracellular trafficking, signal transduction and circulatory stability. A tobacco mosaic virus (TMV) based transient expression system was used to express a single-chain (sc) version of bovine FSH in the tobacco related species Nicotiana benthamiana. Preparations of periplasmic proteins from plants infected with recombinant viral RNA contained high levels of sc-bFSH, up to 3% of total soluble proteins. Consistently, in situ indirect immunofluorescence revealed that the plant cell secreted the mammalian secretory protein to the extracellular compartment (EC). By mass spectrometric analysis of immunoaffinity purified sc-bFSH derived from EC fractions, we found two species of the plant paucimannosidic glycan type, truncated forms of complex-type N glycans. Stimulation of cAMP production in a CHO cell line expressing the porcine FSH receptor acknowledged the native-like structure of sc-bFSH and a sufficient extent of N-glycosylation required for signal transduction. Furthermore, in superovulatory treatments of mice, sc-bFSH displayed significant in vivo bioactivity, although much lower than that of pregnant mare serum gonadotropin. We conclude that plants may have a broad utility as hosts for the recombinant expression of proteins even where glycosylation is essential for function. PMID- 11502220 TI - Essential roles of CD14 and lipopolysaccharide-binding protein for activation of toll-like receptor (TLR)2 as well as TLR4 Reconstitution of TLR2- and TLR4 activation by distinguishable ligands in LPS preparations. AB - Although genetic studies have revealed a critical role for the toll-like receptor (TLR) 4 in the biological response to lipopolysaccharide (LPS), the activities of ectopically expressed TLR4 and TLR2 are controversial. We have found that under appropriate transfection conditions, both TLR2 and TLR4 mediate LPS-induced NF kappaB activation in human embryonic kidney 293 cells. The reconstitution systems we established here allow direct biochemical characterization and comparison of activation of each receptor. TLR4 is approximately 100-fold more sensitive to LPS than TLR2. In contrast to the response to commercial LPS preparations, TLR2 is unresponsive to repurified LPS or synthetic lipid A, indicating the requirement for an additional molecule(s). On the other hand, a lipid A-neutralizing reagent, polymyxin B, blocks the ability of the LPS preparation to stimulate both receptors, suggesting that lipid A is also involved in the activation of TLR2. Mutant TLRs harboring a point mutation in the cytoplasmic domain is inactive in transducing the signal upon stimulation, and act as dominant-negative mutants specifically inhibiting the activation of corresponding type of the receptor but not the other type. Thus, the two receptors are independently activated by distinguishable ligands. Nevertheless, the responses of both TLRs to the LPS preparation are strongly dependent on serum and CD14 and LPS-binding protein are essential for the activation of both of the two receptors. Supporting its functional significance, both receptors are found to associate with CD14. PMID- 11502221 TI - Identification of the beta-dystroglycan binding epitope within the C-terminal region of alpha-dystroglycan. AB - Dystroglycan is a receptor for extracellular matrix proteins that plays a crucial role during embryogenesis in addition to adult tissue stabilization. A precursor product of a single gene is post-translationally cleaved to form two different subunits, alpha and beta. The extracellular alpha-dystroglycan is a membrane associated, highly glycosylated protein that binds to various extracellular matrix molecules, whereas the transmembrane beta-dystroglycan binds, via its cytosolic domain, to dystrophin and many other proteins. alpha- and beta Dystroglycan interact tightly but noncovalently. We have previously shown that the N-terminal region of beta-dystroglycan, beta-DG(654-750), binds to the C terminal region of murine alpha-dystroglycan independently from glycosylation. Preparing a series of deleted recombinant fragments and using solid-phase binding assays, the C-terminal sequence of alpha-dystroglycan containing the binding epitope for beta-dystroglycan has been defined more precisely. We found that a region of 36 amino acids, from position 550-585, is required for binding the extracellular region, amino acids 654-750 of beta-dystroglycan. Recently, a dystroglycan-like gene was identified in Drosophila that showed a moderate degree of conservation with vertebrate dystroglycan (31% identity, 48% similarity). Surprisingly, the Drosophila sequence contains a region showing a higher degree of identity and conservation (45% and 66%) that coincides with the 550-585 sequence of vertebrate alpha-dystroglycan. We have expressed this Drosophila dystroglycan fragment and measured its binding to the extracellular region of vertebrate (murine) beta-dystroglycan (Kd = 6 +/- 1 microM). These data confirm the proper identification of the beta-dystroglycan binding epitope and stress the importance of this region during evolution. This finding might help the rational design of dystroglycan-specific binding drugs, that could have important biomedical applications. PMID- 11502223 TI - Glucose and mental performance. PMID- 11502225 TI - Statistical analysis of nutritional studies. AB - Statistical analysis of data collected in experimental or observational studies is an important part of nutritional research. If it is not done appropriately, there is a risk that information in the data is lost, or that conclusions are misleading. The present article does not attempt a full review of the subject of statistics, but attempts to list aspects where care is needed. PMID- 11502224 TI - Uncoupling proteins: their roles in adaptive thermogenesis and substrate metabolism reconsidered. AB - During the past few years, there have been two major developments, if not revolutions, in the field of energy balance and weight regulation. The first at the molecular level, which was catalysed by developments in DNA screening technology together with the mapping of the human genome, has been the tremendous advances made in the identification of molecules that play a role in the control of food intake and metabolic rate. The second, at the systemic level, which centered upon the use of modern technologies or more robust analytical techniques for assessing human energy expenditure in response to starvation and overfeeding, has been the publication of several papers providing strong evidence that adaptive thermogenesis plays a much more important role in the regulation of body weight and body composition than previously thought. Within these same few years, several new members of the mitochondrial carrier protein family have been identified in a variety of tissues and organs. All apparently possess uncoupling properties in genetically-modified systems, with two of them (uncoupling protein (UCP) 2 and UCP3) being expressed in adipose tissues and skeletal muscles, which are generally recognised as important sites for variations in thermogenesis and/or in substrate oxidation. Considered as breakthrough discoveries, the cloning of these genes has generated considerable optimism for rapid advances in our molecular understanding of adaptive thermogenesis, and for the identification of new targets for pharmacological management of obesity and cachexia. The present paper traces first, from a historical perspective, the landmark events in the field of thermogenesis that led to the identification of these genes encoding candidate UCP, and then addresses the controversies and on-going debate about their physiological importance in adaptive thermogenesis, in lipid oxidation or in oxidative stress. The general conclusion is that UCP2 and UCP3 may have distinct primary functions, with UCP3 implicated in regulating the flux of lipid substrates across the mitochondria and UCP2 in the control of mitochondrial generation of reactive oxygen species. The distinct functions of these two UCP1 homologues have been incorporated in a conceptual model to illustrate how UCP2 and UCP3 may act in concert in the overall regulation of lipid oxidation concomitant to the prevention of lipid-induced oxidative damage. PMID- 11502226 TI - Dexamethasone induces sodium-dependant vitamin C transporter in a mouse osteoblastic cell line MC3T3-E1. AB - The regulation of intracellular ascorbic acid (AsA) levels may be under the control of an AsA-specific membrane transporter. The present study investigates AsA uptake and expression of Na-dependent vitamin C transporter (SVCT) mRNA in the mouse osteoblastic cell line, MC3T3-E1. Among eight compounds tested, dexamethasone (Dex) all-trans retinoic acid, transforming growth factor beta, prostaglandin E2 and transferrin significantly and respectively) stimulated the update of AsA into MC3T3-E1 cells. Among these five, Dex was the most active, inducing mSVCT2 mRNA and the uptake of AsA in a time- and concentration-dependant manner. Dex did not induce mSVCT1 mRNA. These results suggest that the Dex induced stimulation of AsA incorporation into osteoblastic cells is mediated by the induction of mSVCT2. Since Dex reduced alkaline phosphatase activity in MC3T3 E1 cells in our culture conditions, Dex-induced stimulation of AsA incorporation might not be the result of differentiation. Hormone-regulated changes of SVCT expression may have an important role in cell functions. PMID- 11502227 TI - Effects of dietary sucrose on hippocampal serotonin release: a microdialysis study in the freely-moving rat. AB - The effects of dietary supplementation with either sucrose or starch (50 g/kg regular food for 2 weeks) on central 5-hydroxytryptamine (5HT; serotonin) release were investigated in freely-moving rats. It has been suggested that the amount of transmitter that serotoninergic neurons release might be altered by food intake. We monitored the effects of sucrose and starch on concentrations of extracellular 5HT, its metabolite 5-hydroxyindoleacetic acid (5HIAA), gamma-aminobutyric acid (GABA) and dopamine in the hippocampus, using in vivo microdialysis. The major finding was that baseline levels of extracellular hippocampal 5HT in rats with ad libitum access to food supplemented with sucrose were significantly higher compared with the starch control group. We then verified that sucrose supplementation affected the potency of S(+)fenfluramine to increase hippocampal 5HT levels. In both groups of rats, acute intraperitoneal injection (1 mg/kg) of this anorectic drug induced a response curve of the extracellular hippocampal 5HT levels, with a shape that corresponded with earlier data for different brain areas often using up to 10-fold higher doses of S(+)fenfluramine. Nevertheless, we showed that throughout the experiment the absolute values of the sucrose response curve remained higher than in the starch group. On the other hand, S(+)fenfluramine exerted longer lasting effects in the starch group, as compared with the sucrose group. Significant decreases in levels of extracellular hippocampal 5HIAA levels following S(+)fenfluramine administration were simultaneously observed. A practical implication of the present findings is that dietary sucrose may bias the results of studies investigating brain serotoninergic mechanisms and the effects of (anorectic) drugs interacting with 5HT systems in the hippocampus. PMID- 11502228 TI - Reduced thymocyte proliferation but not increased apoptosis as a possible cause of thymus atrophy in iron-deficient mice. AB - Iron deficiency induces thymus atrophy in laboratory animals and very likely in humans by unknown mechanisms. The atrophy is associated with impaired cell mediated immunity. In this study, we tested the hypothesis that thymus atrophy is a result of increased apoptosis and reduced thymocyte proliferation. Thymocytes were obtained from twenty-seven control, twenty-seven pairfed, twenty-seven iron deficient (ID) mice; twelve and fourteen ID mice that received the control diet (0.9 mmol/kg versus 0.09 mmol/kg for the ID diet) for 1 d (repletion, R1) and 3 d (R3), respectively. Cell cycle analysis and apoptosis were studied by flow cytometry using propidium iodide staining and terminal deoxyuridine nick end labeling of DNA breaks assay respectively. When mice were killed, haemoglobin, haematocrit, and liver iron stores of ID, R1, and R3 mice were 25-40 % of those of control and pairfed mice Absolute and relative thymus weights and thymocyte numbers were 19 to 68 % lower in ID, R1, and R3 than in control and pairfed groups We found no significant difference among groups in the percentage of cells undergoing apoptosis. A higher percentage of thymocytes from ID and R1 mice than those of control, pairfed, and R3 mice were in the resting phase of the normal cell cycle Conversely, a lower percentage of thymocytes from ID and R1 mice than those from control, pairfed, and R3 mice were in the DNA synthesis phase and late phase of DNA synthesis and onset of mitosis (G2-M) Indicators of iron status positively correlated (r 0.3 to 0.56) with the percentage of thymocytes in the G2 M phase Results suggest that reduced cell proliferation but not increased apoptosis is the cause of thymus atrophy associated with iron deficiency. PMID- 11502229 TI - Learning behaviour and cerebral protein kinase C, antioxidant status, lipid composition in senescence-accelerated mouse: influence of a phosphatidylcholine vitamin B12 diet. AB - Our objective was to determine whether dietary supplementation with phosphatidylcholine (PC) plus vitamin B12 could afford beneficial effects on biochemical and biophysical events in the brain of senescence-accelerated mouse (SAM) substrain SAMP8. We measured learning behaviour, hippocampal protein kinase C (PKC) activity, cerebral antioxidant status, phospholipid composition and fatty acid composition in 6-month-old SAMP8 and in age-matched controls (SAM substrain SAMR1). In comparison with SAMR1, SAMP8 showed a significant elevation in total grading score of senescence and a significant decline in acquisition SAMP8 had a lower hippocampal PKC activity and cerebral PKC-beta mRNA abundance than SAMR1. SAMP8 had increased cerebral lipid peroxide levels and proportion of sphingomyelin, and a lower proportion of 20 : 4n-6 and 22 : 6n-3 in cerebral phosphtidylethanolamine than SAMR1. SAMP8 fed the PC combined with vitamin B12 diet had an increased PKC activity and a higher proportion of 22 : 6n-3 than SAMP8 fed the control diet. These results indicate the potential benefit of PC combined with vitamin B12 as a dietary supplement. PMID- 11502230 TI - Placebo expectancy effects in the relationship between glucose and cognition. AB - The present study investigated the extent of expectancy in the ability of glucose to affect cognitive performance. Using a within-subjects design, subjects (n 26) completed four experimental sessions (in counterbalanced order and after an initial practice session) during which they were given a 500 ml drink 30 min prior to completing a cognitive assessment battery. In addition, all subjects completed a baseline practice session during which they were given no drink. During two of the sessions, subjects were given a drink containing 50 g glucose and on the other two they were given a drink containing aspartame. A balanced placebo design was used, such that for half the sessions subjects were accurately informed as to the content of the drink (glucose or aspartame), whereas in the other two sessions they were misinformed as to the content of the drink. The task battery comprised a 6 min visual analogue of the Bakan vigilance task, an immediate verbal free-recall task, an immediate verbal recognition memory task and a measure of motor speed (two-finger tapping). Blood glucose and self reported mood were also recorded at several time points during each session. Glucose administration was found to improve recognition memory times, in direct contrast to previous findings in the literature. Glucose administration also improved performance on the Bakan task (relative to the control drink), but only in sessions where subjects were informed that they would receive glucose and not when they were told that they would receive aspartame. There were no effects either of the nature of the drink or expectancy on the other measures. These results are interpreted in terms of there being some contribution of expectancy concerning the positive effects of glucose on cognition in studies which have not used an equi-sweet dose of aspartame as a control drink. PMID- 11502231 TI - Flow-mediated vasodilation is not impaired when HDL-cholesterol is lowered by substituting carbohydrates for monounsaturated fat. AB - Low-fat diets, in which carbohydrates replace some of the fat, decrease serum cholesterol. This decrease is due to decreases in LDL-cholesterol but in part to possibly harmful decreases in HDL-cholesterol. High-oil diets, in which oils rich in monounsaturated fat replace some of the saturated fat, decrease serum cholesterol mainly through LDL-cholesterol. We used these two diets to investigate whether a change in HDL-cholesterol would change flow-mediated vasodilation, a marker of endothelial function. We fed thirty-two healthy volunteers two controlled diets in a weeks' randomised cross-over design to eliminate variation in changes due to differences between subjects. The low-fat diet contained 59.7 % energy (en%) as carbohydrates and 25.7 en% as fat (7.8 en% as monounsaturates); the oil-rich diet contained 37.8 en% as carbohydrates and 44.4 en% as fat (19.3 en% as monounsaturates). Average (sd) serum HDL-cholesterol after the low-fat diet was 0.21 (sd 0.12) mmol/l (8.1 mg/dl) lower than after the oil-rich diet. Serum triacylglycerols were 0.22 (sd 0.28) mmol/l (19.5 mg/dl) higher after the low-fat diet than after the oil-rich diet. Serum LDL and homocysteine concentrations remained stable. Flow-mediated vasodilation was 4.8 (SD 2.9) after the low-fat diet and 4.1 (SD 2.7) after the oil-rich diet (difference 0.7 %; 95 % CI -0.6, 1.9). Thus, although the low-fat diet produced a lower HDL-cholesterol than the high-oil diet, flow-mediated vasodilation, an early marker of cardiovascular disease, was not impaired. PMID- 11502232 TI - Red wine raises plasma HDL and preserves long-chain polyunsaturated fatty acids in rat kidney and erythrocytes. AB - The effects of red wine and ethanol on plasma lipoproteins and the fatty acid composition of kidney lipids and erythrocytes phospholipids were studied. Lipid peroxidation is one of the main deleterious effects of oxidant attack on biomolecules, due to the disruption of the structural integrity of membranes. The vulnerability of the kidney to oxidative damage has been partly attributed to its high content of long-chain polyunsaturated fatty acids. Antioxidants, such as flavonoids, would be a means of reducing the risk of oxidative damage to membranes. Nutritional sources rich in antioxidants, including those provided by wine, are expected to attenuate the effects of oxidative challenges. Adult rats were fed red wine rich in flavonols, ethanol (125 ml/l), or alcohol-free red wine. The control group drank water. After 10 weeks, blood samples served to measure plasma lipoproteins and antioxidant capacity. Kidney lipids and erythrocyte phospholipids were extracted. The samples were assayed by GLC. Energy intake did not differ between all the groups, but the weight gain of the ethanol group was less than the other three groups. Blood HDL and triacylglycerols were increased by both ethanol and red wine. Ethanol decreased arachidonic and docosahexaenoic acids in both kidney lipids and erythrocyte phospholipids, as compared with either water, red wine or alcohol-free red wine groups. These results indicate that non-alcoholic components of red wine could contribute to avoiding the unfavourable effects of ethanol on plasma lipoproteins, kidney lipids and membrane erythrocyte phospholipids. PMID- 11502233 TI - Exercise prevents the augmentation of postprandial lipaemia attributable to a low fat high-carbohydrate diet. AB - There is concern that replacement of dietary fat with carbohydrate may not reduce the overall risk of CHD because this replacement strategy elevates postprandial plasma triacylglycerol (TAG) concentrations. The present study was designed to test the hypothesis that daily exercise can offset the augmented postprandial lipaemia associated with a short-term high-carbohydrate diet. Nine healthy, normolipidaemic men aged 33 (sd 4) years consumed a test meal (g/kg body mass; 1.2 fat, 1.1 carbohydrate, 0.2 protein) on three occasions: after 3 d on a typical Western diet (46, 38 and 16 % energy from carbohydrate, fat and protein respectively); after 3 d on an isoenergetic high-carbohydrate diet (corresponding values: 70, 15 and 15 % energy); after 3 d on the same high-carbohydrate diet with 30 min moderate exercise daily. Fasting plasma TAG concentration was higher after the high-carbohydrate diet (1.15 (se 0.16) mmol/l) than after the Western diet (0.83 (se 0.10) mmol/l; ). Similarly, postprandial lipaemia (6 h total area under plasma TAG concentration v. time curve) was higher after the high carbohydrate diet (12.54 (se 2.07) mmol/l.h) than after the Western diet (9.30 (se 1.30) mmol/l.h; ). The addition of exercise to the high-carbohydrate diet significantly reduced postprandial lipaemia (9.95 (se 1.94) mmol/l.h; when compared with the high-carbohydrate diet) but not fasting TAG concentration (1.02 (se 0.24) mmol/l). In conclusion, daily exercise prevented the augmentation of postprandial lipaemia attributable to the short-term high-carbohydrate diet and, thus, exercise may be a powerful adjunct to dietary change. PMID- 11502234 TI - Effect of diets rich in oleic acid, stearic acid and linoleic acid on postprandial haemostatic factors in young healthy men. AB - The aim of the present study was to investigate the effects of stearic acid-, oleic acid- and linoleic acid-rich meals on postprandial haemostasis in young healthy volunteers whose background diets had been controlled for 14 d in a residential study. Six healthy male volunteers were assigned randomly to consume diets rich in stearic acid, oleic acid or linoleic acid for 14 d. On day 15, plasma lipids and haematological variables were measured in the fasted state, and 3 and 7 h (factor VII and prothrombin activation peptide fragments, 1 and 2 only) after consumption of a test meal. Test meals provided 40 % of the subjects' daily energy requirement, with 41 % of the energy provided as fat, 17 % energy as protein and 42 % energy as carbohydrate. The mean fat content of the meal was 45 (sd 5) g. Significant alterations from fasted values were observed for activated factor VII after 7 h), factor VII antigen after 7 h), prothrombin activation peptide fragments 1 and 2 after 7 h) and plasminogen activator inhibitor type 1 activity after 3 h) after consumption of each of the three meals. No significant differences were observed in haemostatic values (factor VII coagulant activity, factor VII antigen, tissue plasminogen activator activity prothrombin activation peptide fragment and plasminogen activator inhibitor type-1) with regard to diet except for activated factor VII at 3 h; values were higher after the oleic acid- and linoleic acid-rich meals than after the stearic acid-rich meal After consumption of each of the three meals, chylomicrons contained proportionately more palmitic acid than the lipids ingested. The present study shows that there are demonstrable changes in postprandial haemostasis when young healthy volunteers with controlled dietary backgrounds are challenged with a physiological fat load. These changes are independent of the fatty acid composition of the test meals. PMID- 11502235 TI - Dietary phosphate-dependent growth is not mediated by changes in plasma phosphate concentration. AB - The present study was undertaken in order to assess the role of dietary phosphate in growth. A diet deficient in phosphate led to a suppression of growth in juvenile rats. The phenomenon is similar to that described for diets deficient in other essential single components such as Mg, Zn or K. However, unlike the other constituents, dietary phosphate restriction affected the growth rate much more than it altered the serum phosphate concentration; addition of phosphate to the drinking water of rats fed a low-phosphate diet restored the growth rate without a concomitant change in serum phosphate concentration. The suppression of growth rate by the deletion of phosphate was associated with a delayed decrease in food intake. This finding implies that the variation in appetite was secondary to the change in growth. The increase in body weight following phosphate supplementation was associated with a concomitant increment in food intake. The phosphate dependent growth was, however, evident also in rats that were pair-fed with those that were not supplied with phosphate. It is concluded that dietary phosphate dependent growth is not mediated by changes in phosphate concentrations in the extracellular fluid. It is plausible that signals arising from receptors for phosphate in the digestive system constitute part of the growth control apparatus in rats. PMID- 11502236 TI - Phytate levels in diverse rat tissues: influence of dietary phytate. AB - Phytate (inositol hexaphosphate; InsP6) was determined in rat tissues fed on diets with different phytate contents, using a GC-mass detection methodology that permitted the evaluation of the total amount of this substance present in such tissues. The highest InsP6 concentrations were found in brain 5.89 x 10(-2)(SE 5.7 x 10(-3)) mg/g DM), whereas the concentrations detected in kidneys, liver and bone were similar to each other 1.96 x 10(-3) (SE 0.20 x 10(-3), 3.11 x 10(-3) (SE 0.24 x 10(-3), 1.77 x 10(-3) (SE 0.17 x 10(-3)) mg/g DM respectively) and 10 fold less than those detected in brain. When rats were fed on a purified diet in which InsP6 was undetectable, the InsP6 levels of the organs mentioned earlier decreased dramatically (9.0 x 10(-4), 3.8 x 10(-5), 1.4 x 10(-5) mg/g DM in brain, kidneys and liver respectively) and in some cases became undetectable (bone). The addition of InsP6 to this purified diet led to the increase of InsP6 levels in these tissues. This clearly demonstrated that the majority of the InsP6 found in organs and tissues has a dietary origin and is not a consequence of endogenous synthesis. Consequently, considering that InsP6 could be involved in some important biological roles, the value of any diet on supplying this substance is noteworthy. PMID- 11502237 TI - Effects of yoghurt enriched with plant sterols on serum lipids in patients with moderate hypercholesterolaemia. AB - The objective of the present study was to assess the effect of consumption of a yoghurt-based drink enriched with 1-2 g plant sterols/d on serum lipids, transaminases, vitamins and hormone status in patients with primary moderate hypercholesterolaemia. Thirty patients were randomly assigned to one of two treatment groups: a low-fat low-lactose yoghurt-based drink enriched with 1 g plant sterol extracted from soyabean/d v. a low-fat low-lactose yoghurt, for a period of 4 weeks. After a 2-week wash-out period, patients were crossed over for an additional 4-week period. Second, after a 4-week wash-out period, eleven patients were treated with 2 g plant sterols/d in a second open part of the study for a period of 8 weeks. The yoghurt enriched with plant sterols significantly reduced, in a dose-dependent manner, serum total cholesterol and LDL-cholesterol levels and LDL-cholesterol:HDL-cholesterol (P<0.001), whereas no changes were observed in HDL-cholesterol and triacylglycerol levels, either in the first or the second part of the study. There were only slight, not statistically significant, differences in serum transaminase, vitamin and hormone levels. To conclude, a low-fat yoghurt-based drink moderately enriched with plant sterols may lower total cholesterol and LDL-cholesterol effectively in patients with primary moderate hypercholesterolaemia. PMID- 11502238 TI - In vitro availability of zinc from infant foods with increasing phytic acid contents. AB - An in vitro method was used to determine the availability of Zn from infant foods containing increasing amounts of phytate, and to quantify the effect of the phytate:Zn molar ratio on the availability. During the in vitro assay, digestive conditions of infants, younger and older than 4 months of age, were carefully simulated since the solubility of phytate-Zn complexes during digestion is pH dependent. Availability was measured with a continuous flow dialysis in vitro procedure with previous intralumen digestive stage. Zn concentrations were determined with flame atomic absorption spectrometry. Phytic acid content was measured with HPLC. Adding phytate to infant formula lowered Zn availability to 2.84 (sd 0.17) % when the phytate:Zn molar ratio increased to 2.2 as compared with cows' milk-based formula (6.65 (sd 0.55) %). Availability from vegetables (23.83 (sd 2.17) %) significantly decreased at a ratio > 7.9 (15.12 (sd 1.63) %). Zn availability from soyabean-based formula (2.26 (sd 0.36) %) was lower compared with cows' milk-based formula (6.65 (sd 0.55) %). Availability between soyabean- and cows' milk-based formula was similar when a phytate:Zn ratio of 2.2 (2.84 (sd 0.17) %) was obtained in the cows' milk formula. The negative effect of phytic acid on Zn availability was dependent on the type of the food and the phytate content, and should be considered when using soyabean-based formulas during early infancy. PMID- 11502239 TI - Accumulation and apparent oxidation of cis,trans-18 : 2 isomers relative to linoleic acid in rats. AB - Dietary cis,trans-18 : 2 isomers impair desaturation and elongation of linoleic acid (Delta9cis,12cis-18 : 2), but little is known of their proportional partitioning between accumulation and oxidation. The present study was therefore designed to assess the accumulation and apparent oxidation of cis,trans-18 : 2 isomers compared with that of trans-18 : 1 isomers and Delta9cis,12cis-18 : 2 in rats. Accumulation is defined as whole-body increase in a fatty acid during a given period (i.e. final body content-initial body content). The apparent oxidation (disappearance) is defined as whole-body utilization of a fatty acid relative to its intake for a given period (intake-excretion-accumulation-longer chain products)/intakex100). The animals were fed on a diet containing 15 % (w/w) partially hydrogenated rapeseed oil with 1.72 % energy as cis,trans-18 : 2 isomers and varying amounts of Delta9cis,12cis-18 : 2. The apparent oxidation of total cis,trans-18 : 2 isomers (72-76 % dietary intake) was greater than that of Delta9cis,12cis-18 : 2 (38-51 % dietary intake) but it was similar to that of total trans-18 : 1 isomers (78-82 % dietary intake). Among the four isomers, the apparent oxidation of Delta9trans,12trans-18 : 2 was greater than that of the other isomers including Delta9trans,12cis-18 : 2, Delta9cis,12trans-18 : 2 and Delta9cis,13trans-18 : 2. Accumulation of Delta5cis,8cis,11cis,15trans-20 : 4 and Delta5cis,8cis,11cis,14trans-20 : 4 derived from chain-elongation and desaturation of Delta9cis,13trans-18 : 2 and Delta9cis,12trans-18 : 2 was decreased when the dietary Delta9cis,12cis-18 : 2 supply was increased. PMID- 11502240 TI - The role of gut tissue in the energy metabolism of growing lambs fed forage or concentrate diets. AB - The role of the gastrointestinal tract (GIT) in explaining the less efficient utilization of metabolizable energy (ME) in growing lambs fed forage rather than concentrate-based diets was investigated by feeding forage (legume-grass silage) and concentrate (whole shelled maize) diets, at isoenergetic intakes (ME basis), using five groups of lambs. One group of seven lambs was an initial slaughter group and of the two groups (eight lambs per group) fed each diet, one group was fed for 8 weeks, whereas the other group was fed for 16 weeks. All lambs were slaughtered between 18.5 and 20 h following their last meal. Retained energy (as a percentage of ME intake) was higher (concentrate-fed 28, forage-fed 17; P<0.001) for the concentrate-fed animals. Weight-specific mucosal O2 uptake (ml/g DM per h), measured in vitro, was 37 % higher for the forestomach (reticulum, rumen and omasum) and small intestine (jejunum) than for the abomasum and large intestine (caecum and colon), but there was no evidence for a diet effect (except colon; forage-fed 5.3, concentrate-fed 4.2; P=0.036). Total GIT heat loss was estimated as 14 (forage-fed) and 18 (concentrate-fed) % of the whole-body heat loss. Although the GIT did not contribute to increased thermogenesis in the forage-fed lambs in the present study, greater relative contribution of GIT tissue to whole-body mass, i.e. GIT as a percentage of empty-body weight(forage 7.6, concentrate 6.6; P<0.001) in the forage-fed animals supports a role for the GIT in contributing to higher thermogenesis observed in ruminants fed forage as opposed to concentrate diets. PMID- 11502241 TI - Quantifying and separating the effects of macronutrient composition and non macronutrients on energy density. AB - The purpose of the present study was to estimate and compare the effects of macronutrient composition (relative portions of macronutrients) and of non macronutrient components (e.g. water and fibre) on energy density (energy per unit weight) of the diets of human subjects. We used standard macronutrient energy content values to develop a simple conceptual model and equation for energy density in terms of % energy from dietary fat and % non-macronutrients by weight. To study these effects in self-selected diets of free-living subjects, we used four consecutive days of self-weighed and recorded food records for thirty two male and thirteen female free-living adult subjects. In the range of typical human diets, the effect of % non-macronutrients by weight was several times greater than that of % energy from dietary fat, both in absolute terms and relative to daily variation in subjects' diets. Both effects were large enough to be physiologically important. Non-macronutrients (% by weight) alone explained much more of the variation in self-selected dietary energy density either between subjects (R2 95 %) or day-to-day (R2 95 %) than did % energy from dietary fat (R2 5 % and 6 % respectively). Omitting beverages gave similar results. The smaller effect of macronutrient composition on energy density of diets is mainly because alterations in macronutrient composition affect only the portion of typical dietary intake that is macronutrients (one-quarter to one-third of weight). Mathematical methods are also useful in analysing observational data and for separating effects of macronutrient composition and non-macronutrients in intervention studies. These results illustrate the importance of considering non macronutrients in the design and analysis of experimental or observational dietary data. PMID- 11502242 TI - Obesity and undernutrition in a very-low-income population in the city of Maceio, northeastern Brazil. AB - Obesity is the nutritional disorder which has shown the greatest increase in prevalence, even in those countries in which deficiency diseases represent a severe public health problem. The goal of the present study was to analyse the anthropometric profile of a community living in the outskirts of Maceio, capital of Alagoas (northeastern Brazil), and to investigate the hypothesis of a coexistence of undernutrition and obesity in a very low-income population. The survey was conducted on 315 families (1247 individuals). Among the children (aged < or =10 years), the prevalence of wasting, stunting and wasting plus stunting was 3.8, 8.3 and 8.7 % respectively. Wasting (10.2 %) was the most prevalent form of undernutrition among adolescents; nonetheless, a higher frequency of stunting (11 %) and overweight-obesity (5.5 %) was seen specifically in girls, in agreement with trends found in other studies. Adults exhibited a high prevalence of overweight-obesity (25 %), but stunting was also present (22 %). Of the stunted individuals, 30 % were overweight-obese and 16.3 % were underweight. There were eighty-six families with at least one parent who was underweight (27 %) and 104 families with at least one parent who was overweight (33 %). Underweight and overweight-obesity were both present in ninety-six households (30 %). These results may indicate that better living conditions in urban areas in a population 'adapted' to chronic famine might increase the susceptibility to obesity. Considering the harm caused by the cumulative effect of these two conditions (undernutrition in childhood and obesity in adult life) there is a clear need for new studies to uncover the determinant factors so that preventive measures can be implemented. PMID- 11502243 TI - Viability and dose-response studies on the effects of the immunoenhancing lactic acid bacterium Lactobacillus rhamnosus in mice. AB - Previous studies have indicated that the lactic acid bacterium Lactobacillus rhamnosus HN001 can enhance immune function in mice, following oral delivery. However, the influence of bacterial cell viability on immunoenhancement, and the optimum dose of HN001 required for this effect, have not been determined. In the present study, both live and heat-killed preparations of L. rhamnosus HN001 were shown to enhance the phagocytic activity of blood and peritoneal leucocytes in mice, at a dose of 109 micro-organisms daily. In contrast, only live HN001 enhanced gut mucosal antibody responses to cholera toxin vaccine. Feeding mice with 107 viable HN001/d for 14 d was shown to enhance the phagocytic capacity of blood leucocytes, with incremental enhancement observed at 109 and 1011 daily doses. In contrast, a minimum dose of 109 viable HN001/d was required to enhance the phagocytic activity of peritoneal leucocytes, and no further increment was observed with 1011 daily. This study demonstrates that L. rhamnosus HN001 exhibits dose-dependent effects on the phagocytic defence system of mice, and suggests that while the innate cellular immune system is responsive to killed forms of food-borne bacteria, specific gut mucosal immunity may only be stimulated by live forms. PMID- 11502244 TI - Oligofructose and long-chain inulin: influence on the gut microbial ecology of rats associated with a human faecal flora. AB - Dietary incorporation of fermentable, indigestible fructans may be of benefit to gastrointestinal health by providing short-chain fatty acids, stimulating the proliferation of bifidobacteria or lactobacilli and suppressing potential pathogenic organisms in the gut. We tested the hypothesis that the effects of fructans on caecal, colonic and faecal short-chain fatty acid concentration and microflora composition depend on their chain length. Germ-free rats associated with a human faecal flora were randomly assigned to one of four treatments as follows: (1) commercial standard diet as a control (Con); (2) Con+50 g short chain oligofructose/kg (OF); (3) C+50 g long-chain inulin/kg (lcIN); or (4) Con+50 g OF-lcIN/kg (Mix OF-lcIN). Changes in bacterial population groups in response to feeding these diets were investigated with 16S rRNA-targeted probes applied in in situ hybridization. Mix OF-lcIN- and lcIN-containing diets resulted in larger numbers of caecal, colonic and faecal bacteria of the Clostridium coccoides-Eubacterium rectale cluster than Con (10.6 and 10.3 v. 9.5 log10/g wet wt), whereas OF alone did not affect this bacterial group in caecum, colon or faeces. A bifidogenic effect was only observed in the colon and faeces of OF treated rats. More lactobacilli were found in caecal and colonic contents of Mix OF-lcIN-fed rats and in faeces of OF-fed rats compared with Con. Mix OF-lcIN and OF led to significantly smaller numbers of caecal, colonic and faecal bacteria belonging to the Clostridium histolyticum and C. lituseburense groups than Con (6.8 and 6.9 v. 7.9 log10/g wet wt). Counts of total bacteria, Bacteroides Prevotella and Enterobacteriaceae did not differ between the groups. OF and/or lcIN-containing diets significantly increased the caecal and colonic concentration of butyrate and its relative molar proportion. Only lcIN-containing diets resulted in a higher faecal concentration of butyrate than Con. Higher molar proportions of faecal butyrate were observed with all diets that had been supplemented with OF and/or lcIN. Stimulation of butyrate production could be of interest for the prevention of ulcerative colitis and colon cancer. PMID- 11502245 TI - Pneumococcal pulmonary infection, septicaemia and survival in young zinc-depleted mice. AB - The aim of the present study was to explore whether mice fed a diet low in Zn (2.0 mg Zn/kg diet) for a relatively short period of time were more prone to severe Streptococcus pneumoniae infection than mice fed a normal diet (25 mg elemental Zn/kg). The Zn-deficient mice were compared with mice in two Zn adequate control groups; one pair-fed and another with free access to the diet. After 2 weeks feeding, the mice were infected intranasally under anaesthesia with a suspension containing about 10(7) pneumococci. Clinical status was observed every day and blood samples were examined for S. pneumoniae every second day for a week. All infected mice examined carried the infecting strain intranasally. The survival time and time before positive blood culture were significantly shorter in the Zn-depleted group than in the pair-fed Zn-adequate group (hazard ratios 15.6 and 3.2, and respectively). At the end of the observation period, ten of the twelve mice in the Zn-deficient group were dead while one of twelve and two of twelve were dead in the two Zn-adequate control groups. This study shows that even acutely-induced Zn deficiency dramatically increases the risk of serious pneumococcal infection in mice. PMID- 11502246 TI - Energy expenditure by de novo lipogenesis. PMID- 11502247 TI - New building blocks for the dendritic spine. AB - Two recent studies by Sheng and associates (Pak et al., 2001; Sala et al., 2001) provide an elegant molecular analysis of the role of a spine-specific protein, SPAR, and the synaptic proteins Shank and Homer, in regulating dendritic spine morphology, and the possible functional consequences of this regulation. PMID- 11502248 TI - A new critical period for sensory map plasticity. AB - The development of neural circuits is influenced by sensory experience during restricted critical periods early in life. A novel critical period is demonstrated for plasticity of the whisker map in layer 2/3 of rat primary somatosensory cortex. Sensory experience during this period guides initial formation of whisker receptive fields. PMID- 11502249 TI - Tactile shape processing. AB - Neuroimaging techniques may aid in the identification of areas of the human brain that are involved in tactile shape perception. Bodegard et al. (2001) relate differences in the properties of tactile stimuli to differences in areas of cortical activation to infer tactile processing in the somatosensory network. PMID- 11502250 TI - The role of tangential migration in the establishment of mammalian cortex. AB - In the developing nervous system, neurons are generated at sites distant from their ultimate location. In the vertebrate CNS, neurons utilize distinct migration strategies to reach their ultimate residence. This review discusses the contribution of tangential migration to the architectural development of the cerebral cortex and cerebellum. PMID- 11502251 TI - Viktor Hamburger (1900-2001). Journey of a neuroembryologist to the end of the millennium and beyond. PMID- 11502252 TI - A model of synaptic memory: a CaMKII/PP1 switch that potentiates transmission by organizing an AMPA receptor anchoring assembly. AB - Ca2+/calmodulin-dependent protein kinase II (CaMKII) is localized in the postsynaptic density (PSD) and is necessary for LTP induction. Much has been learned about the autophosphorylation of CaMKII and its dephosphorylation by PSD protein phosphatase-1 (PP1). Here, we show how the CaMKII/PP1 system could function as an energy-efficient, bistable switch that could be activated during LTP induction and remain active despite protein turnover. We also suggest how recently discovered binding interactions could provide a structural readout mechanism: the autophosphorylated state of CaMKII binds tightly to the NMDAR and forms, through CaMKII-actinin-actin-(4.1/SAP97) linkages, additional sites for anchoring AMPARs at synapses. The proposed model has substantial experimental support and elucidates principles by which a local protein complex could produce stable information storage and readout. PMID- 11502253 TI - Crossregulation between Neurogenin2 and pathways specifying neuronal identity in the spinal cord. AB - We have examined how genetic pathways that specify neuronal identity and regulate neurogenesis interface in the vertebrate neural tube. Here, we demonstrate that expression of the proneural gene Neurogenin2 (Ngn2) in the ventral spinal cord results from the modular activity of three enhancers active in distinct progenitor domains, suggesting that Ngn2 expression is controlled by dorsoventral patterning signals. Consistent with this hypothesis, Ngn2 enhancer activity is dependent on the function of Pax6, a homeodomain factor involved in specifying the identity of ventral spinal cord progenitors. Moreover, we show that Ngn2 is required for the correct expression of Pax6 and several homeodomain proteins expressed in defined neuronal populations. Thus, neuronal differentiation involves crossregulatory interactions between a bHLH-driven program of neurogenesis and genetic pathways specifying progenitor and neuronal identity in the spinal cord. PMID- 11502254 TI - Crossinhibitory activities of Ngn1 and Math1 allow specification of distinct dorsal interneurons. AB - Distinct classes of neurons are generated from progenitor cells distributed in characteristic dorsoventral patterns in the developing spinal neural tube. We define restricted neural progenitor populations by the discrete, nonoverlapping expression of Ngn1, Math1, and Mash1. Crossinhibition between these bHLH factors is demonstrated and provides a mechanism for the generation of discrete bHLH expression domains. This precise control of bHLH factor expression is essential for proper neural development since as demonstrated in both loss- and gain-of function experiments, expression of Math1 or Ngn1 in dorsal progenitor cells determines whether LH2A/B- or dorsal Lim1/2-expressing interneurons will develop. Together, the data suggest that although Math1 and Ngn1 appear to be redundant with respect to neurogenesis, they have distinct functions in specifying neuronal subtype in the dorsal neural tube. PMID- 11502255 TI - Signaling in channel/enzyme multimers: ATPase transitions in SUR module gate ATP sensitive K+ conductance. AB - ATP-sensitive potassium (K(ATP)) channels are bifunctional multimers assembled by an ion conductor and a sulfonylurea receptor (SUR) ATPase. Sensitive to ATP/ADP, K(ATP) channels are vital metabolic sensors. However, channel regulation by competitive ATP/ADP binding would require oscillations in intracellular nucleotides incompatible with cell survival. We found that channel behavior is determined by the ATPase-driven engagement of SUR into discrete conformations. Capture of the SUR catalytic cycle in prehydrolytic states facilitated pore closure, while recruitment of posthydrolytic intermediates translated in pore opening. In the cell, channel openers stabilized posthydrolytic states promoting K(ATP) channel activation. Nucleotide exchange between intrinsic ATPase and ATP/ADP-scavenging systems defined the lifetimes of specific SUR conformations gating K(ATP) channels. Signal transduction through the catalytic module provides a paradigm for channel/enzyme operation and integrates membrane excitability with metabolic cascades. PMID- 11502256 TI - The expression of vesicular glutamate transporters defines two classes of excitatory synapse. AB - The quantal release of glutamate depends on its transport into synaptic vesicles. Recent work has shown that a protein previously implicated in the uptake of inorganic phosphate across the plasma membrane catalyzes glutamate uptake by synaptic vesicles. However, only a subset of glutamate neurons expresses this vesicular glutamate transporter (VGLUT1). We now report that excitatory neurons lacking VGLUT1 express a closely related protein that has also been implicated in phosphate transport. Like VGLUT1, this protein localizes to synaptic vesicles and functions as a vesicular glutamate transporter (VGLUT2). The complementary expression of VGLUT1 and 2 defines two distinct classes of excitatory synapse. PMID- 11502257 TI - Neurotrophin-induced transport of a beta-actin mRNP complex increases beta-actin levels and stimulates growth cone motility. AB - Neurotrophin regulation of actin-dependent changes in growth cone motility may depend on the signaling of beta-actin mRNA transport. Formation of an RNP complex between the beta-actin mRNA zipcode sequence and Zipcode Binding Protein 1 (ZBP1) was required for its localization to growth cones. Antisense oligonucleotides to the zipcode inhibited formation of this RNP complex in vitro and the neurotrophin induced localization of beta-actin mRNA and ZBP1 granules. Live cell imaging of neurons transfected with EGFP-ZBP1 revealed fast, bidirectional movements of granules in neurites that were inhibited by antisense treatment, as visualized by FRAP analysis. NT-3 stimulation of beta-actin protein localization was dependent on the 3'UTR and inhibited by antisense treatment. Growth cones exhibited impaired motility in the presense of antisense. These results suggest a novel mechanism to influence growth cone dynamics involving the regulated transport of mRNA. PMID- 11502258 TI - Polarized dendritic transport and the AP-1 mu1 clathrin adaptor UNC-101 localize odorant receptors to olfactory cilia. AB - Odorant receptors and signaling proteins are localized to sensory cilia on olfactory dendrites. Using a GFP-tagged odorant receptor protein, Caenorhabditis elegans ODR-10, we characterized protein sorting and transport in olfactory neurons in vivo. ODR-10 is transported in rapidly moving dendritic vesicles that shuttle between the cell body and the cilia. Anterograde and retrograde vesicles move at different speeds, suggesting that dendrites have polarized transport mechanisms. Residues immediately after the seventh membrane-spanning domain of ODR-10 are required for localization; these residues are conserved in many G protein-coupled receptors. UNC-101 encodes a mu1 subunit of the AP-1 clathrin adaptor complex. In unc-101 mutants, dendritic vesicles are absent, ODR-10 receptor is evenly distributed over the plasma membrane, and other cilia membrane proteins are also mislocalized, implicating AP-1 in protein sorting to olfactory cilia. PMID- 11502259 TI - Regulation of dendritic spine morphology by SPAR, a PSD-95-associated RapGAP. AB - The PSD-95/SAP90 family of scaffold proteins organizes the postsynaptic density (PSD) and regulates NMDA receptor signaling at excitatory synapses. We report that SPAR, a Rap-specific GTPase-activating protein (RapGAP), interacts with the guanylate kinase-like domain of PSD-95 and forms a complex with PSD-95 and NMDA receptors in brain. In heterologous cells, SPAR reorganizes the actin cytoskeleton and recruits PSD-95 to F-actin. In hippocampal neurons, SPAR localizes to dendritic spines and causes enlargement of spine heads, many of which adopt an irregular appearance with putative multiple synapses. Dominant negative SPAR constructs cause narrowing and elongation of spines. The effects of SPAR on spine morphology depend on the RapGAP and actin-interacting domains, implicating Rap signaling in the regulation of postsynaptic structure. PMID- 11502260 TI - Rapid development and plasticity of layer 2/3 maps in rat barrel cortex in vivo. AB - Cortical synaptic circuitry develops rapidly in the second postnatal week, simultaneous with experience-dependent turnover of dendritic spines. To relate the emergence of sensory maps to synaptogenesis, we recorded synaptic potentials evoked by whisker deflection in layer 2/3 neurons from postnatal day (P) 12 to 20. At P12, synaptic responses were undetectable. Only 2 days later in life (P14), receptive fields had mature organization. Sensory deprivation, if initiated before P14, disrupted receptive field structure. In layer 4, responses and maps were already mature by P12 and insensitive to deprivation, implying that barrel cortex develops from layer 4 to layer 2/3. Thus, P12-14 is a critical period shared by layer 2/3 synapses and their spines, suggesting that spine plasticity is involved in the refinement of maps. PMID- 11502261 TI - Hierarchical processing of tactile shape in the human brain. AB - It is not known exactly which cortical areas compute somatosensory representations of shape. This was investigated using positron emission tomography and cytoarchitectonic mapping. Volunteers discriminated shapes by passive or active touch, brush velocity, edge length, curvature, and roughness. Discrimination of shape by active touch, as opposed to passive touch, activated the right anterior lobe of cerebellum only. Areas 3b and 1 were activated by all stimuli. Area 2 was activated with preference for surface curvature changes and shape stimuli. The anterior part of the supramarginal gyrus (ASM) and the cortex lining the intraparietal sulcus (IPA) were activated by active and passive shape discrimination, but not by other mechanical stimuli. We suggest, based on these findings, that somatosensory representations of shape are computed by areas 3b, 1, 2, IPA, and ASM in this hierarchical fashion. PMID- 11502262 TI - Recovering meaning: left prefrontal cortex guides controlled semantic retrieval. AB - Prefrontal cortex plays a central role in mnemonic control, with left inferior prefrontal cortex (LIPC) mediating control of semantic knowledge. One prominent theory posits that LIPC does not mediate semantic retrieval per se, but rather subserves the selection of task-relevant knowledge from amidst competing knowledge. The present event-related fMRI study provides evidence for an alternative hypothesis: LIPC guides controlled semantic retrieval irrespective of whether retrieval requires selection against competing representations. With selection demands held constant, LIPC activation increased with semantic retrieval demands and with the level of control required during retrieval. LIPC mediates a top-down bias signal that is recruited to the extent that the recovery of meaning demands controlled retrieval. Selection may reflect a specific instantiation of this mechanism. PMID- 11502263 TI - Preparation of 1,2-O-isopropylidene derivatives of alpha-D-galactoseptanose, beta L-altroseptanose, and 3-O-methyl-alpha-D-guloseptanose. AB - Displacement of the tosyloxy group in 5-O-benzyl-1,2-O-isopropylidene-4-O-(p toluenesulfonyl)-alpha-D-glucoseptanose has yielded derivatives of 1,2-O isopropylidene-alpha-D-galactoseptanose. Acid catalysed acetonation then gave 1,2:3,4-di-O-isopropylidene-alpha-D-galactoseptanose or 1,2;4,5-di-O isopropylidene-alpha-D-galactoseptanose using lower acid concentrations. Reduction of the ketone derived from 1,2:3,4-O-isopropylidene-alpha-D-septanose gave 1,2;3,4-di-O-isopropylidene-beta-L-altroseptanose. Reaction of 3,4-anhydro-5 O-benzyl-1,2-O-isopropylidene-alpha-D-galactoseptanose with sodium methoxide gave 5-O-benzyl-1,2-O-isopropylidene-4-O-methyl-alpha-D-glucoseptanose and 5-O-benzyl 1,2-O-isopropylidene-3-O-methyl-alpha-D-guloseptanose. Solution-state conformations of these compounds have been deduced from their 1H NMR spectra. PMID- 11502264 TI - Complexation of trivalent lanthanide cations by D-ribose in the solid state. The crystal structure and FT-IR study of PrCl3-alpha-D-ribopyranose-5H2O. AB - The crystal structure of praseodymium chloride.alpha-D-ribopyranose pentahydrate, PrCl3-C5H10O5-5 H2O, M(r)=487.47, a=9.1989(8), b=8.8214(7), c=9.8233(9) A, beta=94.060(3) degrees, V=795.2(1) A(3), Z=2, mu=0.71073 A and R=0.0418 for 1923 observed reflections and 172 parameters has been determined. The sugar provides three hydroxyl groups, ax-eq-ax for coordination. The Pr(3+) ion is nine coordinated with five Pr-O bonds from water molecules, three from hydroxyl groups and one from chloride. The OH, CO stretching vibrations and COH bending vibrations are shifted in the complex IR spectrum and the hydroxyl groups, water molecules, chloride ions form an extensive hydrogen-bond network. PMID- 11502265 TI - Synthesis of the milk oligosaccharide 2'-fucosyllactose using recombinant bacterial enzymes. AB - The enzymatic synthesis of GDP-beta-L-fucose and its enzymatic transfer reaction using recombinant enzymes from bacterial sources was examined. The GDP-D-mannose 4,6-dehydratase and the GDP-4-keto-6-deoxy-D-mannose 3,5-epimerase-4-reductase from Escherichia coli K-12, respectively, were used to catalyse the conversion of GDP-alpha-D-mannose to GDP-beta-L-fucose with 78% yield. For the transfer of the L-fucose to an acceptor, we cloned and overproduced the alpha-(1-->2) fucosyltransferase (FucT2) protein from Helicobacter pylori. We were able to synthesise 2'-fucosyllactose using the overproduced FucT2 enzyme, enzymatically synthesised GDP-L-fucose and lactose. The isolation of 2'-fucosyllactose was accomplished by anion-exchange chromatography and gel filtration to give 65% yield. PMID- 11502266 TI - Human ovarian cancer, lymphoma spleen, and bovine milk GlcNAc:beta1,4Gal/GalNAc transferases: two molecular species in ovarian tumor and induction of GalNAcbeta1,4Glc synthesis by alpha-lactalbumin. AB - Affinity Gel-UDP was utilized to purify GlcNAc:beta1,4Gal/GalNAc transferases (Ts) from human lymphoma spleen, ovarian tumor, and ovarian cancer sera. Mn(2+) was found to be an absolute requirement for activity. Two molecular species containing both beta1,4Gal/GalNAc-T activities were discernible when the purified ovarian tumor microsomal enzyme was subjected to Sephacryl S-100 HR column chromatography as well as native polyacylamide gel-electrophoresis. Acceptor specificity studies of the affinity-purified lymphoma spleen and ovarian tumor microsomal enzymes and the conventionally purified, as well as the cloned, bovine milk GlcNAc:beta1,4Gal-Ts using a number of synthetic acceptors showed that the beta(1,6)-linked GlcNAc moiety to alpha-GalNAc was the most efficient acceptor. As compared to the purified milk enzyme, the recombinant form exhibited sixfold GlcNAc:beta1,4 GalNAc-T activity and up to eightfold GlcNAc6SO3beta-:beta1,4Gal-T activity. Further, the recombinant enzyme catalyzed the transfer of GalNAc to the terminal beta-linked GlcNAc6SO3 moiety. Alpha-lactalbumin (alpha-LA) inhibited up to 85%, the transfer of Gal to the GlcNAc moiety linked either to Man or GlcNAc. On the contrary, alpha-LA had no significant influence on the transfer of GalNAc to the above acceptors. alpha-LA had no appreciable effect on the recombinant enzyme, except for the transfer of Gal or GalNAc to Glc. Both alpha- and beta glucosides, as well as alpha-N-acetylglucosaminide, did not serve as acceptors. PMID- 11502267 TI - Investigation of inclusion complexation of paclitaxel by cyclohenicosakis-(1-->2) (beta-D-glucopyranosyl), by cyclic-(1-->2)-beta-D-glucans (cyclosophoraoses), and by cyclomaltoheptaoses (beta-cyclodextrins). AB - Inclusion complexation of the poorly soluble drug, paclitaxel, was investigated with various host cyclooligosaccharides such as a family of isolated neutral cyclohenicosakis-(1-->2)-(beta-D-glucopyranosyl) (cyclic-(1-->2)-beta-D-glucans, cyclosophoraoses), dimethyl cyclomaltoheptaose (cyclodextrins, DM-beta-CD) and hydroxypropyl cyclomaltoheptaose (cyclodextrins, HP-beta-CD). Quantitative analysis with high-performance liquid chromatography (HPLC) indicated that all three cyclic oligosaccharides could increase the solubility of paclitaxel, where DM-beta-CD gave the best results and a family of cyclosophoraoses and HP-beta-CD, both gave similar results. Complexation of host molecules with paclitaxel was studied by NMR and fluorescence spectroscopic analyses. NMR spectroscopic analysis showed that the aromatic regions of paclitaxel experienced noticeable changes of the chemical shifts or peak shapes upon interaction with host molecules. The relatively bulky cyclosophoraoses allowed favorable accessibility to either the B-ring or A-ring of paclitaxel, while DM-beta-CD and HP-beta-CD allowed accessibility to all the aromatic rings including the C ring. The interaction of DM-beta-CD with paclitaxel greatly increased the fluorescence intensity compared with other host molecules, suggesting the more effective partitioning of a moderate fluorophore into a hydrophobic cluster adjacent to the C-ring of paclitaxel. PMID- 11502268 TI - Structural analysis of the exopolysaccharide from Burkholderia caribensis strain MWAP71. AB - Burkholderia caribensis strain MWAP71 was isolated from rhizosphere soil microaggregates in Martinique. The extracellular polysaccharide produced by this strain was found to be composed of D-glucose (D-Glc), 6-deoxy-L-talose (L-6dTal), 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), and an O-acetyl group in a molar ratio of 2:1:1:1. The primary structure of the polysaccharide was shown by sugar analysis, electrospray mass spectrometry, partial acid hydrolysis and 1-D and 2-D NMR spectroscopy to consist of a tetrasaccharide repeating unit having the following structure: [structure in text]. PMID- 11502269 TI - Isolation of oligogalacturonic acids up to DP 20 by preparative high-performance anion-exchange chromatography and pulsed amperometric detection. AB - Milligram quantities of oligogalacturonic acids up to a degree of polymerization (DP) of 20 were purified by high-performance anion-exchange chromatography utilizing a preparative-scale (21-mm i.d.) CarboPac PA1 column and a nonlinear potassium acetate (pH 7.5) gradient. Detection was accomplished by pulsed amperometry without post-column addition of hydroxide. Pulsed amperometry at near neutral pH is an excellent detection method for preparative separations of carbohydrates because it avoids base-catalyzed degradation reactions that can occur at high pH. This method was simpler, faster, had higher sample loading capacity and allowed for the isolation of higher DP oligogalacturonic acids than other methods reported previously. With this improved method, multi-milligram quantities of valuable oligogalacturonic acids (up to DP 20) can be readily isolated. PMID- 11502270 TI - Effect of binding of Calcofluor White on the carbohydrate residues of alpha1-acid glycoprotein (orosomucoid) on the structure and dynamics of the protein moiety. A fluorescence study. AB - Calcofluor White is a fluorescent probe that interacts with polysaccharides and is commonly used in clinical studies. Interaction between Calcofluor White and carbohydrate residues of alpha1-acid glycoprotein (orosomucoid) was previously studied at low and high concentrations of Calcofluor compared to that of the protein. alpha1-Acid glycoprotein contains 40% carbohydrate by weight and has up to 16 sialic acid residues. At equimolar concentrations of Calcofluor and alpha1 acid glycoprotein, the fluorophore displays free motions [Albani, J. R.; Sillen, A.; Coddeville, B.; Plancke, Y. D.; Engelborghs, Y. Carbohydr. Res. 1999, 322, 87 94], while at high concentration of Calcofluor, its surrounding microenvironment is rigid, inducing the rigidity of the fluorophore itself [Albani, J. R.; Sillen, A.; Plancke, Y. D.; Coddeville, B.; Engelborghs, Y. Carbohydr. Res. 2000, 327, 333-340]. In the present work, red-edge excitation spectra and steady-state anisotropy studies performed on Trp residues in the presence of Calcofluor, showed that the apparent dynamics of Trp residues are not modified. However, deconvoluting the emission spectra with two different methods into different components, reveals that the structure of the protein matrix has been disrupted in the presence of high Calcofluor concentrations. PMID- 11502271 TI - Synthesis of S-linked glucosaminyl-4-thiohex-2-enopyranosides via allylic SN2' substitution of a tosylhexenose. AB - Treatment of 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-1-thio-beta-D-glucopyranose with 1,6-anhydro-3,4-dideoxy-2-O-p-toluenesulfonyl-beta-D-erythro-hex-3 enopyranose gave 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-beta-D-glucopyranosyl-(1- >4)-1,6-anhydro-2,3-dideoxy-4-thio-beta-D-erythro-hex-2-enopyranose in 86% yield. Its 1,6-anhydride bond was cleaved with methanol to give a mixture of methyl glycosides (alpha/beta approximately 5:1), from which the alpha anomer was separated by crystallization and converted into its 6-acetate, 6 methanesulfonate, or deacetylated to obtain the corresponding free methyl thiodisaccharide. The structure of the new compounds was confirmed by 1H and 13C NMR spectra. PMID- 11502272 TI - Synthesis of steroidal glycosides bearing the disaccharide moiety of OSW-1 and their antitumor activities. AB - Nine glycosides bearing the disaccharide of OSW-1, namely 2-O-(4-methoxybenzoyl) beta-D-xylopyranosyl-(1-->3)-2-O-acetyl-alpha-L-arabinopyranosides, were synthesized, and their antitumor activities were tested. PMID- 11502273 TI - FK506 promotes adenosine release from endothelial cells via inhibition of adenosine kinase. AB - The immunosuppressants, cyclosporin A and tacrolimus (FK506) induce an increase in plasma levels of adenosine and mimic ischemic preconditioning. However, the mechanism of action of the two drugs on adenosine metabolism is not clear. Since inhibition of adenosine kinase promotes an increase in endogenous adenosine release, we tested a hypothesis that FK506 induces adenosine release via inhibition of adenosine kinase activity. In cultured endothelial cells, FK506 enhanced release of tracer adenosine and inhibited uptake of tracer adenosine. It also reduced adenosine kinase activity of the cell membrane fraction. In addition, FK506 does not inhibit membrane transport of tracer adenosine. These observations indicate that FK506 inhibits in situ adenosine kinase activity in endothelial cells. Other cell signaling inhibitors were found to inhibit adenosine uptake via inhibition of adenosine transport. In conclusion, FK506 promotes adenosine release from endothelial cells by a novel mechanism involving inhibition of adenosine kinase activity associated with the membrane. PMID- 11502274 TI - pH-dependent blocking actions of three novel antiarrhythmic compounds on K+ and Na+ currents in rat ventricular myocytes. AB - Three novel chemically related compounds were studied for their pH-dependent ion channel blocking actions on the transient outward K+ current (I(to)) and the Na+ current (I(Na)) in isolated rat ventricular myocytes. The (+/-)-trans napthylethoxycyclohexylamines, RSD1108, RSD1070 and RSD1067, showed similar potencies in reducing the inactivation time course of I(to) at pH 7.4. However, RSD1108 (pK(a) 6.8) was a more potent blocker of I(to) at pH 6.4 than the other two compounds (pK(a) values near 8.0). The reduction of inactivation times induced by the RSD compounds was consistent with open channel blockade and in consequence an open channel block model was used in order to estimate blocking and unblocking rate constants. This analysis showed no apparent correlation between pK(a) and onward blocking rate constants for the compounds. However, the unblocking rate constant for the low pK(a) compound RSD1108 at acid pH decreased markedly from that found at normal pH. Both RSD1108 and RSD1070 showed an enhanced potency to block I(Na) at acid pH relative to pH 7.4. However, RSD1108 showed significantly less inhibition of I(Na) at both pH values compared to RSD1070 and RSD1067. Differences in channel block were also evident between RSD1070 and RSD1067, which could be attributed to the difference in napthyl groups between their chemical structures. The compounds exhibited use- and frequency-dependent blockade of I(Na) with the amount of use-dependent blockade greater for RSD1108 and RSD1067 than for RSD1070 at acid pH compared to neutral pH. Greater frequency-dependent inhibition was apparent for RSD1108 as compared to RSD1070 and RSD1067 at both pH 7.4 and 6.4. These results point out the importance of the magnitude of pK(a) and chemical structure in ion channel blocking actions of a series of structurally related compounds. PMID- 11502275 TI - Budesonide reduces vascular endothelial growth factor secretion and expression in airway (Calu-1) and alveolar (A549) epithelial cells. AB - Vascular endothelial growth factor (VEGF), a cytokine expressed in the respiratory epithelial cells, induces vascular hyperpermeability and edema, symptoms that are alleviated by budesonide, an anti-asthma corticosteroid. However, modulation of VEGF levels by budesonide in the respiratory epithelium has not been studied. In this study, we investigated the mechanisms of VEGF secretion using brefeldin A and monensin in human airway (Calu-1) and alveolar (A549) epithelial cells, and further determined whether budesonide inhibits VEGF secretion and mRNA expression through a glucocorticoid receptor-mediated mechanism. In both cell types, VEGF secretion was inhibited by brefeldin A and monensin, suggesting vesicular transport of VEGF through endoplasmic reticulum (ER)-golgi pathway. At concentrations devoid of cytotoxicity, budesonide reduced VEGF secretion and VEGF mRNA expression in both cell types and these effects were inhibited by mifepristone (RU 486), a glucocorticoid receptor antagonist, suggesting that budesonide reduces VEGF secretion and expression through its glucocorticoid receptor-mediated action. Also, budesonide-mediated inhibition of VEGF mRNA was time- and protein synthesis-dependent. Thus, budesonide may be of potential value in treating disorders of the respiratory tract that are associated with VEGF elevation. PMID- 11502276 TI - L-carnitine prevents increase in diastolic [CA2+] induced by doxorubicin in cardiac cells. AB - Doxorubicin is a highly effective anticancer chemotherapeutic agent that produces a dose-dependent cardiomyopathy that limits its clinical usefulness. We investigated the acute effects of doxorubicin on diastolic free Ca2+ concentrations ([Ca2+]) and the cardioprotective action of L-carnitine in isolated cardiomyocytes loaded with fura-2/AM (acetoxymethyl ester). Exposure to 10(-6)-10(-4) M doxorubicin induced an elevation of diastolic calcium concentration ([Ca2+]) that was concentration dependent. Nitrendipine failed to prevent the doxorubicin-induced elevation of [Ca2+]. Incubation with L-carnitine (10(-8) to 10(-6) M) did not modify [Ca2+]. Pretreatment of cardiomyocytes with L carnitine 10(-8)-10(-7) M did not prevent the doxorubicin effect on [Ca2+]. However, L-carnitine 10(-6) M fully inhibited the increase in [Ca2+] induced by this anthracycline derivative. These results indicate that acute exposure to doxorubicin impairs intracellular Ca2+ handling and that L-carnitine exerts a cardioprotective effect, in part by preventing the doxorubicin-induced increase in diastolic Ca2+ concentration. PMID- 11502277 TI - Tolerance to the anticonvulsant activity of midazolam and allopregnanolone in a model of picrotoxin seizures. AB - The effects of an intracerebroventricular (i.c.v.) administration of a non selective full benzodiazepine receptor agonist, midazolam, and a neuroactive steroid, allopregnanolone, on picrotoxin-induced seizures and striatal dopamine metabolism, were studied in mice. It was found that acute i.c.v. injections of midazolam (ED50=38.25 nmol) and allopregnanolone (ED50=26.34 nmol) blocked picrotoxin-induced seizures to a similar extent. After repeated administration at the ED(85) doses (midazolam-56.6 nmol, allopregnanolone-94.2 nmol; once or twice daily for 5 days) tolerance developed to the anticonvulsant activity of midazolam (ED50=94.14 nmol) and allopregnanolone (ED50=186.70 nmol). Acute i.c.v. injections of midazolam and allopregnanolone (at the ED50 doses established in the model of picrotoxin seizures: 38.25 and 26.34 nmol, respectively), significantly decreased the concentration of dopamine metabolites: 3 methoxytyramine and 3,4-dihydroxyphenylacetic acid, as well as the dopamine turnover rate (homovanilic acid/dopamine ratio; by about 20%), in the mouse striatum. These findings together with the recently published data on the potentiation by midazolam and allopregnanolone of ethanol-induced sleep [Pharmacol. Biochem. Behav. 67 (2000) 345] indicate a very similar central effect profile of benzodiazepines and neurosteroids. Moreover, similar efficacy of allopregnanolone and midazolam at the GABA(A) receptors has been found. Overall, the results of the present study, along with the possibility of neurosteroid conversion in the brain into other steroid hormones (testosterone, estradiol, aldosterone), add to the accumulating evidence suggesting a less favorable pharmacological profile for this class of drugs than was previously thought. PMID- 11502278 TI - Lidocaine potentiates atrial natriuretic peptide-induced relaxation of bovine tracheal smooth muscle. AB - The effect of lidocaine on the changes in tension and guanosine 3',5'-cyclic monophosphate (cGMP) content induced by atrial natriuretic peptide (ANP) and nitric oxide (NO) was examined in bovine tracheal smooth muscle preparations contracted with methacholine (0.3 microM). Lidocaine (10 microM) did not affect the methacholine-induced tensions, whereas 100 microM lidocaine significantly (P<0.01) attenuated methacholine-induced ones. Treatment of the tracheal preparations with lidocaine (10 and 100 microM) significantly (P<0.05) augmented the relaxant responses to ANP, whereas the same procedure did not alter the responses to sodium nitroprusside, (+/-)-(E)-ethyl-2-[(E)-hydroxyimino]-5-nitro-3 hexeneamide (NOR 3) or 8-bromo-cGMP. Lidocaine (100 microM) enhanced cGMP accumulation induced by ANP (0.1 microM) but not by sodium nitroprusside (0.3 microM). In contrast, mexiletine (100 microM), another class Ib antiarrhythmic, did not affect ANP- and sodium nitroprusside-induced relaxations. These results suggest that lidocaine augments ANP-induced relaxation and cGMP accumulation, probably by modulating activation mechanism of particulate guanylyl cyclase. PMID- 11502279 TI - Melatonin protects against renal oxidative stress after obstructive jaundice in rats. AB - The goals of this study were to analyze the renal oxidative status in experimental biliary obstruction and to evaluate the impact of melatonin on renal oxidative stress. Cholestasis was done by double ligature and section of the extra-hepatic biliary duct. Melatonin was injected i.p. (500 microg/kg/day). Malondialdehyde, reduced glutathione, catalase, superoxide dismutase, glutathione reductase, glutathione peroxidase and glutathione transferase were determined in the renal tissue. After biliary obstruction, an increase in malondialdehyde (P<0.0001) and a fall in reduced glutathione (P<0.0001) were seen. Moreover, the scavenger enzyme activity had significantly diminished. After melatonin administration, the malondialdehyde fell significantly (P<0.0001), whereas reduced glutathione showed an important increase (P<0.0001) compared with the ligated bile duct group. Experimental bile duct obstruction was associated to an increase of renal oxidative stress. Treatment with melatonin decreased the renal lipid peroxidation, and both the reduced glutathione as well as the scavenger enzyme activity recovered. PMID- 11502280 TI - Alleviation of wood smoke-induced lung injury by tachykinin receptor antagonist and hydroxyl radical scavenger in guinea pigs. AB - We recently reported that wood smoke inhalation initially (within 5 min) causes airway injury and subsequently produces both airway and parenchymal injury after a delay (within 2 h). In this study, we investigated the mediator mechanisms of this delayed smoke-induced lung injury in 126 anesthetized and artificially ventilated guinea pigs who received challenges of either air or 40 tidal breaths of wood smoke. Two hours after inhalation, wood smoke produced various injurious responses, including increases in alveolar-capillary permeability, microvascular permeabilities, and histological injury scores, in airway and parenchymal tissues. Pre-treatment given before smoke challenge with CP-96,345 [a tachykinin NK1 receptor antagonist; (2S,3S)-cis-2-(diphenylmethyl)-N-((2-methoxyphenyl) methyl)-1-aza bicyclo(2.2.2.)-octan-3-amine], dimethylthiourea (a hydroxyl radical scavenger), or a combination of these two drugs largely alleviated both the airway and parenchymal responses, whereas pre-treatment with SR-48,968 [a tachykinin NK2 receptor antagonist; (S)-N-methyl-N(4-(4-acetylamino-4 phenylpiperidino)-2-(3,4-dichlorophenyl)-butyl)benzamide] or a combination of CP 96,344 and SR-48,965 (inactive enantiomers) failed to do so. Post-treatment given at 5 min after smoke challenge with CP-96,345 or dimethylthiourea significantly alleviated the parenchymal responses, while having no effect on the airway responses. Pre-treatment with dimethylthiourea prevented the smoke-induced reduction in airway neutral endopeptidase activity (an enzyme for tachykinin degradation). We concluded that (1) tachykinins and hydroxyl radical play important roles in producing smoke-induced delayed lung injury in guinea pigs, and both may be involved in the spread of injury from the airways to the pulmonary parenchyma, and (2) the contribution of tachykinins is mediated via the activation of tachykinin NK1 receptors, and is associated with the hydroxyl radical-induced inactivation of airway neutral endopeptidase. PMID- 11502281 TI - Experimental meningitis in the rat: protection by uric acid at human physiological blood concentrations. AB - The natural peroxynitrite scavenger uric acid was previously shown to be protective in a rat model of pneumococcal meningitis; however, rats have much lower blood uric acid levels than humans. Therefore, we evaluated its therapeutic effect at human physiological blood concentrations. Intraperitoneal pretreatment with uric acid increased its blood concentrations from 44.9+/-10.0 microM in untreated rats to 169.8+/-122.6 microM and reduced the cerebrospinal fluid (CSF) pleocytosis from 12767+/-2520 to 8376+/-2450 cells/microl (P<0.05) and the intracranial pressure from 11.6+/-3.0 to 4.3+/-1.2 mm Hg (P<0.05). Coadministration of oxonic acid, an inhibitor of urate oxidase, increased the blood uric acid levels to 355.0+/-79.6 microM and further reduced the CSF pleocytosis (4190+/-1749 cells/microl, P<0.05) and the intracranial pressure (1.4+/-2.4 mm Hg). Uric acid+oxonic acid also had a beneficial effect when administered 2 or 4 h after the induction of meningitis. We demonstrate a dose dependent anti-inflammatory effect of uric acid at blood levels in the human physiological range. PMID- 11502282 TI - Inhibition of TPA-induced cyclooxygenase-2 expression and skin inflammation in mice by wogonin, a plant flavone from Scutellaria radix. AB - Wogonin (5,7-dihydroxy-8-methoxyflavone), isolated from Scutellaria radix, was previously reported to inhibit the expression and activity of the enzyme cyclooxygenase-2 in lipopolysaccharide (LPS)-stimulated cells of a mouse macrophage cell line, RAW 264.7. Here, in order to find in vivo effects, inhibition by wogonin of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cyclooxygenase-2 expression and anti-inflammatory activity in vivo were investigated. When applied topically to the dorsal skin of mice, wogonin at doses of 50-200 microg/site/treatment (total of five treatments in 3 days) inhibited cyclooxygenase-2 expression and prostaglandin E2 production induced by multiple treatments with TPA. At 200 microg/site/treatment, wogonin caused a 55.3% reduction of prostaglandin E2 production on the dorsal skin compared with an increased production in the TPA-treated control group. The same compound significantly inhibited mouse ear edema induced by TPA in both preventive (58.1% inhibition) as well as curative treatment (31.3% inhibition) schedules at 200 microg/ear/treatment. Inhibition of neutrophil infiltration was also observed. Therefore, wogonin may be beneficial for cyclooxygenase-2-related skin disorders. PMID- 11502285 TI - The AMDD program: history, focus and structure. AB - The AMDD (Averting Maternal Death and Disability) Program was established at the Mailman School of Public Health in 1999. In this article, we discuss four key aspects of the program: the focus on emergency obstetric care; the use of process indicators; working with partners; and applying human rights. PMID- 11502283 TI - Expression of preprotachykinin-B, the gene that encodes neurokinin B, in the rat uterus. AB - Neurokinin B, a peptide belonging to the tachykinin family, is undetectable in peripheral tissues from nonpregnant animals. In the present study, we analysed the expression of the preprotachykinin-B (PPT-B) gene, which encodes neurokinin B, in the rat uterus. Preprotachykinin-B mRNA was expressed in the uterus and its levels varied greatly depending upon the hormonal conditions. This is consistent with a role of this tachykinin in the regulation of uterine functions. PMID- 11502287 TI - Monitoring obstetric services: putting the UN guidelines into practice in Malawi. I: developing the system. AB - The UNICEF/WHO/ UNFPA "Guidelines for Monitoring the Availability and Use of Obstetric Services" was published in 1997 as a guide for implementing process indicators. The Malawi Safe Motherhood Project covers 5 million people and was the first large project to introduce the new indicators as part of a routine monitoring system. A rigorous process of needs assessment, tools development, operations research, field testing and training was conducted. It was necessary to reach agreement on locally appropriate standard definitions of obstetric emergencies, reporting formats and catchment populations. Underreporting of emergencies, misreporting of maternal deaths and double counting of referrals were minimized by improving recording tools. Time, cost, political and technical inputs are important considerations--in Malawi, the system took 1 year to set up at a cost of $100 000. Developing a routine monitoring system to obtain data for the process indicators is feasible, but requires adaptation of the cross sectional evaluation methods described in the UN Guidelines. PMID- 11502289 TI - Holding up a mirror: changing obstetric practice through criterion-based clinical audit in developing countries. AB - The objective of the study described is to assess the feasibility and effectiveness of using a criterion-based clinical audit to measure and improve the quality of obstetric care at the district hospital level in developing countries. The focus is on the management of five life-threatening obstetric complications--hemorrhage, eclampsia, genital tract infection, obstructed labor and uterine rupture was audited using a "before and after" design. The five steps of the audit cycle were followed: establish criteria of good quality care; measure current practice (Review I); feedback findings and set targets; take action to change practice; and re-evaluate practice (Review II). Systematic literature review, panel discussions and pilot work led to the development of 31 audit criteria. Review I included 555 life-threatening complications occurring over 66 hospital-months; Review II included 342 complications over 42 hospital months. Many common areas for improvement were identified across the four hospitals. Agreed mechanisms for achieving these improvements included clinical protocols, reviews of staffing, and training workshops. Some aspects of clinical monitoring, drug use and record keeping improved significantly between Reviews I and II. Criterion-based clinical audit in four typical district hospitals in Ghana and Jamaica is a feasible and acceptable method for quality assurance and appears to have improved the management of life-threatening obstetric complications. PMID- 11502291 TI - Malaria prophylaxis and the reduction of anemia at childbirth. AB - OBJECTIVES: To assess the effect of introducing chloroquine prophylaxis during pregnancy on prevalence of anemia (<10.9 g/dl) at childbirth and perinatal outcome. METHODS: Observational study in a rural district hospital in Ghana, which compared 2803 women who received chloroquine prophylaxis during pregnancy with 3084 historical controls, who had not received prophylaxis during pregnancy. Main outcome measures were hemoglobin level at childbirth, perinatal mortality and birth weight. RESULTS: Mean hemoglobin level before childbirth increased from 10.7 g/dl (S.D.=1.0 g/dl) to 11.0 g/dl (S.D.=0.9 g/dl). Prevalence of anemia decreased from 29.4 to 13.3% (OR=0.4 and 95% CI=0.3-0.4). Prevalence of moderately severe anemia (<9.0 g/dl) decreased from 4.4 to 3.3% (OR=0.7, 95% CI=0.6-0.97). Perinatal mortality and low birth weight (<2500 g) remained unchanged. CONCLUSIONS: Routine chloroquine prophylaxis in pregnancy is useful in reducing anemia at childbirth in malaria-endemic regions. Fetal outcome did not improve with chloroquine prophylaxis in this study. PMID- 11502292 TI - Tamponade-balloon for obstetrical bleeding. AB - OBJECTIVES: To study the effectiveness of a large volume, fluid-filled tamponade balloon in the management of post-partum hemorrhage originating from the implantation site of low-lying placenta/placenta previa. METHODS: A silicone, fluid-filled balloon was designed for tamponade function, with a filling capacity volume of 500 cc of sterile saline, and strength to withstand a maximum internal and external pressure of 300 mmHg. Five women with postpartum bleeding caused by low-lying placenta/placenta previa, and one woman with cervical ectopic pregnancy, underwent a tamponade balloon insertion as a conservative measure in the management of bleeding. RESULTS: The tamponade balloon was used in five women with post-partum bleeding caused by low-lying placenta/placenta previa, and in one woman with cervical pregnancy. The balloon was effective in controlling post partum hemorrhage originating from the placental site of the lower uterine segment, and bleeding from the implantation site of cervical ectopic pregnancy. CONCLUSION: Hemostasis in cases of post-partum bleeding caused by low-lying placenta/placenta previa can be achieved by using a large volume, fluid-filled tamponade balloon. PMID- 11502293 TI - Delivery of patients with early onset, severe pre-eclampsia. AB - OBJECTIVES: To compare the effects of induction/labor to delivery before labor in early onset, severe pre-eclampsia. METHODS: Five-year prospective case series. Delivery course and neonatal outcome were examined for 335 women with viable singletons. RESULTS: Induction was successful in 45% of attempts. Women exposed to labor had longer (5.5 days, P<0.0001) admissions to delivery periods and were more often delivered for maternal indications (RR=2.87, 95% CI=1.98-4.16). Their babies were born 1.6 weeks older (P<0.0001) and 352 g heavier (P<0.0001) than those delivered before labor. Babies exposed to labor needed intensive care less often (RR=0.4, 95% CI=0.27-0.58), had lower rates of severe hyaline membrane disease (RR=0.26, 95% CI=0.11-0.59) and sepsis (RR=0.56, 95% CI=0.33-0.93), and were discharged earlier (P<0.0001). CONCLUSIONS: Exposure to induction/labor in selected patients is not detrimental to neonatal outcome in early, severe pre eclampsia. PMID- 11502294 TI - Induction of labor in breech presentation. AB - OBJECTIVE: Induction of labor in breech presentation, although not contraindicated, has rarely been reported. We have undertaken to evaluate the safety and outcome of this practice in two Israeli institutions along with a literature review of this controversial subject. METHOD: The research design was a retrospective case control study covering the years: 1980-1999. We have studied 53 term (>37 weeks) breech deliveries induced for various medical and obstetrical reasons, in two major regional hospitals in Israel. Induction was performed with prostaglandin E(2) for the unripe cervix and with oxytocin for induction or augmentation when the cervix was ripe. Six women were induced by nipple stimulation. Controls were 53 women with spontaneous labor in breech presentation that had a trial of vaginal delivery, and 54 women with breech presentation who delivered by elective cesarean section. RESULT: No significant difference in the various maternal and fetal outcomes was observed. CS rate was comparable in both study and control groups (34% vs. 32%) and two-thirds gave birth vaginally. CONCLUSIONS: In properly selected and carefully managed cases of breech presentation, induction of labor seems a safe and reasonable option. PMID- 11502295 TI - Computerized analysis of fetal heart rate parameters by gestational age. AB - OBJECTIVE: The aim of this study is to define the reference ranges for fetal heart rate (FHR) parameters according to gestational age, by determining the relationship between the FHR and gestational age using a computerized FHR analysis system. METHODS: Using our own software developed by Hanyang University Hospital in Korea, non-stress tests were performed for 20 min. FHR parameters for 6455 subjects were analyzed for various gestational groups; <25 weeks, 25-28 weeks, 29-32 weeks, 33-36 weeks, 37-40 weeks, and >40 weeks. RESULTS: The FHR parameters were related to gestational age. The mean baseline FHR, signal loss, and fetal movements decreased significantly with gestation (P<0.0001). The variability and accelerations of FHR were highest for the 37-40 weeks gestational group (P<0.05). CONCLUSIONS: The results showed that overall, the differences in the FHR parameters between gestational groups were statistically significant, and the gestational age of the fetus should be considered when interpreting FHR patterns. PMID- 11502296 TI - Transvaginal sacrospinous colpopexy for marked uterovaginal and vault prolapse. AB - OBJECTIVE: The transvaginal sacrospinous ligament fixation technique was used as part of the vaginal repair procedure for marked uterovaginal prolapse, and in the treatment of vault prolapse. METHOD: Out of the 26 women treated with sacrospinous ligament suspension of the vaginal vault, 23 had marked uterovaginal prolapse and three had vault prolapse following hysterectomy. Patients with vault prolapse underwent posterior vaginal repair, obliteration of the enterocele sac and sacrospinous colpopexy. Patients with marked uterovaginal prolapse underwent vaginal hysterectomy with high ligation of the enterocele sac, anterior and posterior vaginal repair, and sacrospinous colpopexy. Bilateral salpingoopherectomy was added to the procedure in five patients. All patients were examined 6 weeks after the operation and, subsequently, on an annual basis. The mean follow-up period was 2.6 years (1-5 years). RESULTS: Out of the three patients with previous vault prolapse, none had recurrences. Out of the 23 patients with previous marked uterovaginal prolapse, only two had small cystocele, and one had small enterocele at 36 months following the operation. These patients were asymptomatic and did not need an operation. Vaginal vault prolapse was not observed in any of these patients. Two women had post-operative urinary tract infection and five had buttock discomfort, which subsided after 2 months. No other intra- or post-operative complications occurred. CONCLUSION: Transvaginal sacrospinous colpopexy can be performed together with vaginal hysterectomy, and anterior and posterior vaginal wall repair in patients with marked uterovaginal prolapse because of its high success in avoiding possible vault prolapse, and low intra- and post-operative complication rates. PMID- 11502297 TI - Dystocia: a study of its frequency and risk factors in seven cities of west Africa. AB - OBJECTIVES: To determine the incidence of dystocia in seven west African cities, to attempt to discover what, if any, factors at the prenatal visit might identify women at risk of dystocia, and to assess the utility of such screening. METHOD: This prospective population study of 20326 pregnant women in west Africa (MOMA) analyzed risk factors for dystocia on the basis of deliveries in health care facilities. RESULTS: Incidence of dystocia was 18.3%. In the multivariate analysis, the risk factors were small stature, previous cesarean, and nulliparity. As screening tools these factors have inadequate positive predictive values, either singly or combined. CONCLUSION: It is almost impossible to predict the occurrence of dystocia before the onset of labor. Therefore, labor must be carefully monitored, and there must be health care facilities available that can manage complications, especially cesarean deliveries. If such facilities are not accessible, an effective referral system must be established. PMID- 11502298 TI - A community based study on the change of practice of female genital mutilation in a Sudanese village. AB - OBJECTIVE: To investigate the practice of female genital mutilation (FGM), among young and old parents. METHODS: One hundred and twenty young parents and grandparents in a rural area in central Sudan were randomly selected for interviews carried out according to structured questionnaires with open answer possibilities. RESULTS: All female respondents had undergone FGM. Of the young respondents, 44% had decided not to let their daughters undergo FGM. Young fathers were more involved in the decision process than previously known, especially when decisions were taken not to perform FGM. Tradition and social pressure were the main motives for performing FGM. Sexuality was an important aspect, mentioned both as motives for and against FGM. Religious belief and education level significantly affected to what extent FGM was practiced. CONCLUSION: This is the first community based study of FGM indicating a significant shift in practice between generations, young parents starting to question the value of FGM. PMID- 11502299 TI - Assisted reproduction developments in the Islamic world. AB - A November 2000 workshop organized by the International Islamic Center for Population Studies and Research, Al-Azhar University, Cairo, considered use of assisted reproduction technologies (ART) in the Islamic world. The workshop reinforced a 1997 recommendation that a Standing Committee for Shari'a Medical Ethics be constituted to monitor and assess developments in ART practice. Among issues the workshop addressed were equitable access to services for infertile couples of modest means, and regulation of standards of equipment and personnel that ART centers should satisfy to gain approval to offer services. Acceptable uses of preimplantation genetic diagnosis were proposed, and follicular maturation research in animals, including in vitro maturation and in vitro growth of oocytes, was encouraged, leading to human applications. Embryo implantation following a husband's death, induced postmenopausal pregnancy, uterine transplantation and gene therapy were addressed and human reproductive cloning condemned, but cloning human embryos for stem cell research was considered acceptable. PMID- 11502300 TI - Subclinical chorioamnionitis as a causal factor in unexplained stillbirths. PMID- 11502301 TI - The impact of an intensive care unit on maternal mortality. PMID- 11502302 TI - Effects of progestogen-only contraceptives on breast-feeding and infant growth. PMID- 11502303 TI - A randomized trial of interferon-alpha in cervical dysplasia. PMID- 11502304 TI - Outcome of failed pregnancies managed in an early pregnancy unit. PMID- 11502305 TI - Delivery of retained placenta by misoprostol in second trimester abortion. PMID- 11502306 TI - Children and EtOH--the dangers need spelling out. PMID- 11502307 TI - Desire for physician-assisted suicide: requests for a better death? PMID- 11502308 TI - Derangement of apoptosis in cancer. PMID- 11502309 TI - What is the role of growth-hormone therapy in short children who were small for gestational age? PMID- 11502310 TI - Hot foods--unexpected help with energy balance? PMID- 11502311 TI - Detention and mandatory treatment for tuberculosis patients in Russia. PMID- 11502312 TI - Ombudsman's fifth report. PMID- 11502313 TI - Cholesterol and all-cause mortality in elderly people from the Honolulu Heart Program: a cohort study. AB - BACKGROUND: A generally held belief is that cholesterol concentrations should be kept low to lessen the risk of cardiovascular disease. However, studies of the relation between serum cholesterol and all-cause mortality in elderly people have shown contrasting results. To investigate these discrepancies, we did a longitudinal assessment of changes in both lipid and serum cholesterol concentrations over 20 years, and compared them with mortality. METHODS: Lipid and serum cholesterol concentrations were measured in 3572 Japanese/American men (aged 71-93 years) as part of the Honolulu Heart Program. We compared changes in these concentrations over 20 years with all-cause mortality using three different Cox proportional hazards models. FINDINGS: Mean cholesterol fell significantly with increasing age. Age-adjusted mortality rates were 68.3, 48.9, 41.1, and 43.3 for the first to fourth quartiles of cholesterol concentrations, respectively. Relative risks for mortality were 0.72 (95% CI 0.60-0.87), 0.60 (0.49-0.74), and 0.65 (0.53-0.80), in the second, third, and fourth quartiles, respectively, with quartile 1 as reference. A Cox proportional hazard model assessed changes in cholesterol concentrations between examinations three and four. Only the group with low cholesterol concentration at both examinations had a significant association with mortality (risk ratio 1.64, 95% CI 1.13-2.36). INTERPRETATION: We have been unable to explain our results. These data cast doubt on the scientific justification for lowering cholesterol to very low concentrations (<4.65 mmol/L) in elderly people. PMID- 11502314 TI - Mortality in patients with coeliac disease and their relatives: a cohort study. AB - BACKGROUND: Although previous studies have shown increased mortality in patients with coeliac disease and their relatives, no data are available in relation to different patterns of clinical presentation. We assessed mortality in patients with coeliac disease and their first-degree relatives. METHODS: We enrolled, in a prospective cohort study, 1072 adult patients with coeliac disease consecutively diagnosed in 11 gastroenterology units between 1962 and 1994, and their 3384 first-degree relatives. We compared the number of deaths up to 1998 with expected deaths and expressed the comparison as standardised mortality ratio (SMR) and relative survival ratio. FINDINGS: 53 coeliac patients died compared with 25.9 expected deaths (SMR 2.0 [95% CI 1.5-2.7]). A significant excess of mortality was evident during the first 3 years after diagnosis of coeliac disease and in patients who presented with malabsorption symptoms (2.5 [1.8-3.4]), but not in those diagnosed because of minor symptoms (1.1 [0.5-2.2]) or because of antibody screening (1.2 [0.1-7.0]). SMR increased with increasing delay in diagnosis and for patients with poor compliance with gluten-free diet. Non-Hodgkin lymphoma was the main cause of death. No excess of deaths was recorded in relatives with coeliac disease. INTERPRETATION: Prompt and strict dietary treatment decreases mortality in coeliac patients. Prospective studies are needed to clarify the progression of mild or symptomless coeliac disease and its relation to intestinal lymphoma. PMID- 11502315 TI - Origins of the desire for euthanasia and assisted suicide in people with HIV-1 or AIDS: a qualitative study. AB - BACKGROUND: Euthanasia and assisted suicide, and policies to address them are the subjects of contentious debate in many countries. However, the question of why people desire euthanasia or assisted suicide has not been coherently answered. We aimed to answer this question in a specific group of patients. METHODS: We did a qualitative study of 32 people with HIV-1 or AIDS, who were enrolled in the HIV-1 Ontario Observational Database at Sunnybrook and Women's College Health Sciences Centre, Toronto, Ontario, Canada. We elicited participants' experiences of deliberation about euthanasia or assisted suicide, and the meaning of these experiences with in-depth, face-to-face interviews. We analysed our data with grounded theory methods. FINDINGS: Participants' desire for euthanasia and assisted suicide were affected by two main factors: disintegration, which resulted from symptoms and loss of function; and loss of community, which we defined as progressive diminishment of opportunities to initiate and maintain close personal relationships. These factors resulted in perceived loss of self. Euthanasia and assisted suicide were seen by participants as means of limiting loss of self. INTERPRETATION: These determinants of desire for euthanasia or assisted suicide in people with HIV-1 or AIDS have implications for the debate on these practices, and development of policies to regulate them. PMID- 11502317 TI - Amodiaquine, sulfadoxine/pyrimethamine, and combination therapy for treatment of uncomplicated falciparum malaria in Kampala, Uganda: a randomised trial. AB - BACKGROUND: Increasing Plasmodium falciparum resistance to chloroquine in sub Saharan Africa necessitates use of alternative antimalarial agents. Affordable alternative treatments include sulfadoxine/pyrimethamine and amodiaquine. Combination of antimalarial agents can increase therapeutic efficacy and delay emergence of drug resistance. We compared the efficacy of sulfadoxine/pyrimethamine, amodiaquine, and an amodiaquine/sulfadoxine/pyrimethamine combination for treatment of uncomplicated malaria in a region of high chloroquine resistance. METHODS: Patients with symptoms of uncomplicated falciparum malaria and confirmed disease in Kampala, Uganda, were randomly assigned to receive sulfadoxine/pyrimethamine (25 mg/kg sulfadoxine, and 1.25 mg/kg pyrimethamine) plus placebo; amodiaquine (25 mg/kg) plus placebo; or amodiaquine plus sulfadoxine/pyrimethamine. Patients were followed up for 14 days, and clinical and parasitological outcomes were assessed. FINDINGS: 90% (400/445) of patients enrolled in the study successfully completed 14 days of follow-up. Treatment failure based on clinical criteria occurred in 13 of 131 (10%) patients on sulfadoxine/ pyrimethamine, nine of 131 (7%) on amodiaquine, and four of 138 (3%) on amodiaquine/sulfadoxine/pyrimethamine. Based on parasitological criteria, treatment failed in 26%, 16%, and 10% of these patients, respectively. Amodiaquine/sulfadoxine/pyrimethamine was significantly more effective than sulfadoxine/pyrimethamine alone in children aged younger than 5 years (clinical failure in 3.5% vs 13.9%, respectively, risk difference 10.4% [95% CI, 1.6-19.3] p=0.021; parasitological failure in 12.8% vs 26.4%, risk difference 13.6% [1.2-26.0] p=0.041). INTERPRETATION: Sulfadoxine/pyrimethamine, amodiaquine, and amodiaquine/sulfadoxine/pyrimethamine were all effective for treatment of uncomplicated falciparum malaria in Uganda. The amodiaquine/sulfadoxine/pyrimethamine combination was the most effective, and could be the optimum low-cost alternative to chloroquine in Africa. PMID- 11502318 TI - Monitoring of large randomised clinical trials: a new approach with Bayesian methods. AB - BACKGROUND: In judging whether or not to continue enrolling patients into a randomised clinical trial, most data-monitoring and ethics committees (DMECs) rely on the p value for the difference in effect between the study groups. In the 1990s, two randomised controlled trials-one in patients with lung cancer and one in those with head and neck cancer-were instead monitored by Bayesian methods. We assessed the value of this approach in the monitoring of these clinical trials. METHODS: Before the trials opened, participating clinicians were asked their opinions on the expected difference between the study treatment (continuous hyperfractionated accelerated radiotherapy [CHART]) and conventional radiotherapy. These opinions were used to form an "enthusiastic" and a "sceptical" prior distribution. These prior distributions were combined with the trial data at each of the annual DMEC meetings. If, during monitoring, a result in favour of CHART was seen, the DMEC was to decide whether the results were sufficiently convincing to persuade a sceptic that CHART was worthwhile. Conversely, if there was apparently no or little difference, the DMEC was asked whether they thought the results sufficiently convincing to persuade an enthusiast that CHART was not worthwhile. FINDINGS: At each of the annual meetings, the DMEC concluded that there was insufficient evidence to convert either sceptics or enthusiasts, and that the trials should therefore remain open to recruitment. Neither trial was closed to recruitment earlier than planned. However if a conventional (p-value-based) stopping rule had been used, the lung cancer trial would probably have been stopped. INTERPRETATION: This Bayesian approach to monitoring is simple to implement and straightforward for members of the DMEC to understand. In our opinion, it is more intuitively appealing than conventional approaches. PMID- 11502319 TI - Antibodies that stop the adrenals in their tracks. PMID- 11502320 TI - Frequency of C3435T polymorphism of MDR1 gene in African people. AB - The variability of P-glycoprotein expression between individuals is linked to a C3435T polymorphism of the human MDR1 gene. Concentration of P-glycoprotein in intestinal epithelial cells and in a subset of lymphoid cells is substantially lower in people with the T/T genotype than those with the C/C genotype. We compared allele frequencies of the C3435T polymorphism in random samples of west African, African American, white, and Japanese people. We recorded a significantly higher frequency of the C/C genotype in West Africans and African Americans (142 of 172 [83%] and 25 of 41 [61%], respectively), than in white people (139 of 537 [26%]) (p<0.0001). These findings could affect use of drugs that are P-glycoprotein substrates (such as HIV-1 protease inhibitors and ciclosporin) in African populations. PMID- 11502321 TI - Prediction of severe disseminated adenovirus infection by serum PCR. AB - Adenoviruses are increasingly recognised as viral pathogens that can cause fatal infections in immunocompromised patients, particularly recipients of haematopoietic stem-cell grafts. Adenovirus infections are not easily diagnosed and the development of a severe infection cannot be predicted by standard culture techniques. In a pilot study, we investigated the value of adenovirus DNA detection in serum as a marker of disseminated disease in 14 patients with defined patterns of adenovirus infections. The results show that the appearance of adenoviral DNA in serum preceded the development of a severe or fatal adenovirus infection. Because proper management is dependent on early diagnosis and differentiation from other conditions, this test may be a valuable tool in the management of adenovirus infection. PMID- 11502322 TI - Incidence of childhood acute lymphoblastic leukaemia in Yorkshire, UK. AB - Between 1980 and 1998, in the north-west of England, a significant rise in childhood acute lymphoblastic leukaemia was caused by an increase in the precursor B-cell form of this disease. We analysed data on children who were diagnosed with leukaemia in Yorkshire, UK, between 1974 and 1997. The incidence of acute lymphoblastic leukaemia remained stable, although a non-significant yearly increase of 2.4% was noted for the precursor B-cell form of this disease from 1980 onwards. The precursor B-cell form accounted for 80% of all acute lymphoblastic leukaemia. Our data are not consistent with increasing incidence for precursor B-cell acute lymphoblastic leukaemia, although numbers of children with acute myeloid leukaemia are rising. PMID- 11502323 TI - A case-control study of food poisoning seen at an accident and emergency department. AB - We investigated the hypothesis that sporadic food poisoning is a result of the consumption of food cooked or prepared outside the home. We did a case-control study, set in an urban emergency department, to find out the odds ratios for the risk of food poisoning associated with various patterns of consumption. We found that recent eating out (exposure on the day of or day before presentation) was associated with an odds ratio of 2.41 (95% CI 1.29-4.50) for presenting with food poisoning. PMID- 11502324 TI - US rejection sets biological weapons treaty adrift. PMID- 11502330 TI - Of tardigrades, trehalose, and tissue engineering. PMID- 11502331 TI - Johns Hopkins research returns to normal. PMID- 11502332 TI - Ireland leaves disabled children's right to extended education in doubt. PMID- 11502333 TI - Senior health official in Pakistan arrested on corruption charges. PMID- 11502334 TI - US university and Indian government investigates cancer drug trial. PMID- 11502336 TI - Catalonian government expands patients' right to choose. PMID- 11502337 TI - UK government launches nationwide sexual health strategy. PMID- 11502338 TI - The art and science of clinical knowledge: evidence beyond measures and numbers. AB - Medical doctors claim that their discipline is founded on scientific knowledge. Yet, although the ideas of evidence based medicine are widely accepted, clinical decisions and methods of patient care are based on much more than just the results of controlled experiments. Clinical knowledge consists of interpretive action and interaction-factors that involve communication, opinions, and experiences. The traditional quantitative research methods represent a confined access to clinical knowing, since they incorporate only questions and phenomena that can be controlled, measured, and counted. The tacit knowing of an experienced practitioner should also be investigated, shared, and contested. Qualitative research methods are strategies for the systematic collection, organisation, and interpretation of textual material obtained from talk or observation, which allow the exploration of social events as experienced by individuals in their natural context. Qualitative inquiry could contribute to a broader understanding of medical science. PMID- 11502339 TI - Model of peripheral autonomous modules and a myovesical plexus in normal and overactive bladder function. AB - Normal bladder function is controlled by the central nervous system (CNS) and any peripheral contribution to bladder control is believed to be small. Nevertheless, anatomically and functionally, such a contribution might exist. Taking account of this evidence, we propose that the detrusor muscle is arranged into modules, which are circumscribed areas of muscle active during the filling phase of the micturition cycle. These modules might be controlled by a peripheral myovesical plexus, consisting of intramural ganglia and interstitial cells. Detrusor overactivity is the occurrence of abnormal increases in pressure during bladder filling, which the patient cannot inhibit. This disorder is thought to be a consequence of abnormal expression of the micturition reflex or changes in the properties of the smooth muscle. We propose that detrusor overactivity results from exaggerated symptomatic expression of peripheral autonomous activity, resulting from a shift in the balance of excitation and inhibition in smooth muscle modules. These structures responsible for origin and spread of peripheral autonomous activity could be targeted to help develop new therapeutic strategies. PMID- 11502340 TI - Community-based approaches to HIV treatment in resource-poor settings. PMID- 11502341 TI - Preventing antiretroviral anarchy in sub-Saharan Africa. AB - Combination antiretroviral therapy has dramatically improved the survival of patients living with HIV and AIDS in industrialised countries of the world. Despite this enormous benefit, there are some major problems and obstacles to be overcome.(1) Treatment of HIV-infection is likely to be lifelong.(2) Unfortunately, many HIV-infected individuals cannot tolerate the toxic effects of the drugs, or have difficulty complying with treatment which involves large numbers of pills and complicated dosing schedules. Poor adherence to treatment leads to the emergence of drug-resistant viral strains that need new combinations of drugs or new drugs altogether. PMID- 11502349 TI - Effects of acetylcholine and GABA on neurons in the area postrema of Suncus murinus brainstem slices. AB - Slices (400 microm) containing the area postrema were prepared from three Japanese house musk shrews. Data reported are from 16 representative units. Units fell into one of three separate groups with spontaneous firing rates of 36-41/s, 16-18/sec. and 6-8/sec., possibly reflecting different set points of an endogenous pacemaker. Most neurons did not respond to ionophoresis of GABA or ACh. In those that did, ACh briefly increased spike frequency in a dose-dependent manner. The firing pattern suggests that the membrane potential was depolarized strongly, coupled with an extremely large conductance increase. Bath perfusion of curare, but not atropine, inhibited the response. Ionophoresis of GABA caused strong inhibition. The data show Suncus AP contains neurons that are sensitive to nicotine and GABA and may be emetogenic. PMID- 11502350 TI - Reduction of glial fibrillary acidic protein-immunoreactive astrocytes in some brain areas of old hairless rhino-j mice (hr-rh-j). AB - Mutations in the hairless (hr) gene of mice result in hair follicle and other epithelial defects. The hr gene is expressed at high levels in the brain where it probably participates in the survival and maintenance of some neuronal populations, but whether it also supports glial populations of the central nervous system has been not investigated. To clarify this, quantitative immunohistochemistry for astrocytes (glial fibrillary acidic protein (GFAP)) and microglial cells (CD11b macrophage antigen) was used in the brain of a mutant mouse strain, the hairless (hr-rh-j) type, which carries the homozygous hr gene rhino mutation. The glial cell density was assessed in the cerebral cortex, hippocampus, striatum, hypothalamus and cerebellum of young (3 months) and old (9 months) hr-rh-j mice. No significant differences were found between young wild type and hr-rh-j mice. The density of GFAP immunoreactive astrocytes normally increased as a function of age, but in older hr-rh-j mice there was a severe reduction (P<0.01) in the striatum, hypothalamus, and hippocampus. Conversely, the microglial cells were insensible to aging or to hr-rh-j mutation. These results suggest that the hr gene is involved in the maintenance of the GFAP immunoreactive cells in some cerebral areas. Nevertheless, because these animals do not show any neurological signs, the functional significance of the present findings remains to be established. PMID- 11502351 TI - Dextromethorphan increases tyrosine hydroxylase mRNA in the mesencephalon of adolescent rats. AB - Dextromethorphan (DM), an antitussive widely available in over-the-counter, has been abused mostly in teenage groups at high doses. To examine effects of DM on the reward pathway, we injected a high dose of DM (40 mg/kg; intraperitoneally) into the adolescent rat and measured tyrosine hydroxylase (TH) mRNA by in situ hybridization in the ventral tegmental area (VTA) and the substantia nigra (SN). Remarkable increases in the level of TH mRNA were observed in the VTA and SN 2 h after DM injection. Stereotyped behavior and ataxia increased, and rearing decreased by DM administration. These results suggest that DM-induced increase in TH mRNA expression in mesencephalon contribute to the reinforcing property and the behavioral effects of DM. PMID- 11502352 TI - Tau accumulation in a patient with pallidonigroluysian atrophy. AB - We studied the brain of a patient with pallidonigroluysian atrophy (PNLA) in whom argyrophilic and abnormally phosphorylated tau positive neurons and glia were identified in the brain on Gallyas-Braak silver staining and immunohistochemical analysis although neurofibrillary tangles were not seen by Bodian silver stain. Immunohistochemical studies using six anti-tau antibodies that recognize the different phosphorylated epitopes of tau protein revealed that these epitopes in neurons and glial cells share common characteristics with neurofibrillary tangles in Alzheimer's disease. Immunoblot analysis of phosphorylated tau protein showed major bands of 64 and 68 kDa and after dephosphorylation, tau consisted mainly of 4 repeat tau. No mutations were detected in the coding exons and their flanking intronic regions of the tau gene. This study suggests that PNLA is one of tauopathy and the biochemical characteristics of phosphorylated tau are similar to those found in progressive supranuclear palsy and corticobasal degeneration. PMID- 11502353 TI - Tea catechin, (-)-epigallocatechin gallate, causes membrane depolarizations of myenteric neurons in the guinea-pig small intestine. AB - Intracellular recordings were made from enteric neurons in the myenteric plexus of the guinea-pig small intestine to examine actions of one of tea catechins, (-) epigallocatechin gallate (EGCG), on neuronal activity. EGCG at concentrations from 1 to 20 microM depolarized about 60% of both of S and AH neurons in a concentration-dependent manner, although EGCG-induced depolarizations were largely varied in amplitude from neuron to neuron. The depolarizations started passing off during the presence of EGCG at higher concentrations, and became smaller or almost abolished with repeated applications, indicating the EGCG induced response shows desensitization-like characteristics. The EGCG depolarization was associated with a decrease in input membrane resistance, and their reversal potential was estimated about -30 mV. Since the EGCG depolarization was recorded in the absence of external Ca(2+), it is concluded that one of tea catechins has direct depolarizing effects on a population of myenteric neurons. PMID- 11502354 TI - The spindle neurons are present in the cingulate cortex of chimpanzee fetus. AB - We report the existence of the spindle neurons in layer Vb of the anterior cingulate cortex (Brodman's area 24b) in a chimpanzee fetus (embryonic day 224), which was stillborn. About 5.3% of neuronal cells in layer Vb were the spindle neurons at this stage. The width of the spindle neurons was 10-15 microm. In layer V of the prefrontal cortex (Brodman's area 46) in the chimpanzee fetus, and in layer Vb of the cingulate cortex in adult macaque monkeys, no spindle neurons were observed. The immunoreactivity against brain-derived neurotrophic factor (BDNF) was detected only in the pyramidal neurons at this stage. These findings suggest that the existence of the spindle neurons in layer Vb of the anterior cingulate cortex and the presence of BDNF in the pyramidal neurons are intrinsically characterized during the embryonic stage of the chimpanzee. PMID- 11502355 TI - Synergism between A(2A)-adenosine receptor activation and vasoactive intestinal peptide to facilitate [3H]-acetylcholine release from the rat motor nerve terminals. AB - The effect of vasoactive intestinal peptide (VIP) on evoked [(3)H]-acetylcholine ([(3)H]-ACh) release from motor nerve terminals, and its interaction with presynaptic facilitatory A(2A)-adenosine receptors was investigated in the rat phrenic nerve-hemidiaphragms. Facilitation of [(3)H]-ACh release by VIP (100 nM) only becomes apparent when high frequency (50 Hz) or long lasting pulses (1 ms) were delivered to the phrenic nerve; VIP excitation was prevented by removal of endogenous adenosine tonus, with adenosine deaminase (2.5 units/ml) or with the A(2A)-receptor antagonist, 3,7-dimethyl-1-propargyl xanthine, (10 microM). Pretreatment with the selective A(2A)-receptor agonist, CGS 21680C (2 nM), potentiated the neurofacilitatory action of VIP (100 nM). The results suggest that tonic A(2A)-receptors activation by endogenous adenosine is required to trigger the facilitatory action of VIP on evoked [(3)H]-ACh release from motor nerve endings. PMID- 11502356 TI - Non-linear electroencephalogram dynamics in patients with spontaneous nocturnal migraine attacks. AB - The present study was conducted to examine non-linear electroencephalogram (EEG) measures during the development of a spontaneous migraine attack. We investigated the sleep EEG of five patients with migraine without aura in the pain-free interval and at the onset of a nocturnal attack. Sleep EEG recordings were analysed using the method of global dimensional complexity compared to conventional sleep scoring techniques. We found no divergence between classical sleep architecture and the estimated dimensional course nor any relevant short term changes related to the onset of headache. There was, however, a loss of dimensional complexity in the first two non-rapid eye movement sleep states in the migraine night, with statistical significance during the second sleep cycle. For the first time, these results provide evidence of a global dimension decrease that is related to cortical network changes during a migraine attack. PMID- 11502357 TI - Neural correlates of response inhibition for behavioral regulation in humans assessed by functional magnetic resonance imaging. AB - To identify the specific frontal and cingulate regions involved in response inhibition, five Chinese right-handed male volunteers with an attention span reaching or exceeding a 7-digit level participated in this blocked design functional magnetic resonance imaging study. Each block represented one of the two experimental conditions, the Correct Matching or the Incorrect Matching condition. In the Correct Matching condition, the subjects were required to make correct judgement of whether the two 3-digit numbers presented in succession were the same or not; whereas in the Incorrect Matching condition, the participants were instructed to inhibit the correct responses. Bilateral activations of the prefrontal and anterior cingulate as well as the left posterior cingulated cortex were observed while the subjects were exercising response inhibition. The roles of these activations in response inhibition were discussed. PMID- 11502358 TI - Bromodeoxyuridine and methylazoxymethanol exposure during brain development affects behavior in rats: consideration for a role of nerve growth factor and brain derived neurotrophic factor. AB - Rats prenatally exposed to the neurotoxins methylazoxymethanol (MAM) or 5-Bromo 2'-deoxyuridine (BrdU) are used as animal models of brain maldevelopment. We administered in rats MAM (20 mg/kg), or BrdU (100 mg/kg) or both at gestational day 11. Locomotion was not affected by any prenatal treatment whereas learning was delayed in the Morris maze in MAM animals. BrdU induced decreased NGF and BDNF levels in the hippocampus. In the parietal cortex prenatal BrdU administration induced NGF potentation associated with decreased BDNF. Animals treated with both MAM and BrdU showed also an increased immunopositivity for choline acetyltransferase (ChAT) and low affinity neurotrophins' receptor (p75) in the septum and Meynert's nuclei. These findings suggest that embryonic exposure to MAM and/or BrdU may be useful for studying mechanisms associated with neurodegenerative diseases affecting brain morphology and behavior. PMID- 11502359 TI - Multiple G-protein-coupled receptors mediate presynaptic inhibition at single excitatory synapses in the rat visual cortex. AB - Modulation of excitatory synaptic transmission by agonists for several neurotransmitter receptors was investigated at intrinsic cortical synapses derived from single presynaptic neurons. Excitatory postsynaptic currents (EPSCs) were recorded from layer 5 pyramidal neurons in the rat visual cortex in response to minimal stimulation within the same layer. 5-hydroxytryptamine, adenosine, baclofen, carbachol and DCG-IV all suppressed EPSCs with an increase in paired pulse ratio. These agonists reduced the frequency of miniature EPSCs without significantly affecting their amplitude distribution. These results suggest that glutamatergic excitatory transmission in the neocortex is under the control of presynaptic inhibition mediated by multiple neuromodulator receptors co-expressed in single presynaptic terminals. PMID- 11502360 TI - Time course of recovery of the somatosensory map following hindpaw sensory deprivation in the rat. AB - Hindlimb sensory deprivation is known to induce a decrease in the cortical representation of hindpaw, and an increase in the size of the cutaneous receptive fields. The aim of the present study was to determine (i) the time-course of recovery when the rat retrieves a normal use of its limbs after a 14-day period of sensory disruption and (ii) whether a 1-day period of sensory deprivation is sufficient to induce a plasticity. Our results indicate that the remodelling of the cortical map was not observed after 1 day of sensory deprivation. On the other hand, the recovery was achieved after 6 h. These findings suggest that a procedure reducing sensory function resulted in reversible changes in the somatosensory cortex. The recovery was more rapid than the induction of plasticity. PMID- 11502361 TI - Specific actions of cyanide on membrane potential and voltage-gated ion currents in rostral ventrolateral medulla neurons in rat brainstem slices. AB - The present study examined specific effects of sodium cyanide (CN) on the membrane potential (MP), spontaneous discharge (SD) and voltage-gated ion current of the identified bulbospinal rostral ventrolateral medulla (RVLM) neuron in the rat pup brainstem slice. 125 microM CN rapidly depolarized MP in the RVLM neuron by 11.6 mV as well as enhanced the SD rate by 300%. In contrast, the same dose of CN immediately hyperpolarized unlabeled, non-RVLM neurons by 4.8 mV. 50 microM CN did not significantly affect voltage-gated Ca(++) or A-type K(+) currents. The same concentration of CN, however, rapidly and reversibly suppressed voltage gated Na(+) currents and sustained outward K(+) currents in the RVLM neuron by 22.5% and 23%, respectively. Tetraethylammonium could mimic the effect of CN on MP, SD and sustained K(+) current in the RVLM neuron. It is concluded that: (1) like that from the adult rat, the rat pup bulbospinal RVLM neuron can be selectively and rapidly excited by CN; (2) the hypoxia-sensitive, sustained outward K(+) channel may play an important role in the acute hypoxia-induced excitation of the RVLM neurons. PMID- 11502362 TI - Nerve growth factor induces the expression of the LIM homeodomain transcription factor Isl-1 with the kinetics of an immediate early gene in adult rat dorsal root ganglion. AB - LIM-homeodomain genes encode a major class of transcription factors which play a critical role in regulating tissue specific gene expression. In this report, we have shown that three members of the LIM-homeodomain gene family - Isl-1, Rlim and Lim-3 are expressed in adult rat sensory neurons. Furthermore, we show that the addition of nerve growth factor (NGF) to cultures of primary dorsal root ganglion neurons leads to the induction of Isl-1, Rlim and Lim-3 mRNA expression. The increase in Isl-1 mRNA levels upon NGF addition was rapid and occurred even in the presence of cycloheximide. These findings place Isl-1 as a novel member of the immediate early class of genes. In contrast, Rlim and Lim-3mRNA induction by NGF required protein synthesis. The role of LIM-homeodomain genes in mediating responses to NGF in adult sensory neurons is discussed. PMID- 11502363 TI - Association study between novel promoter variants at the 5-HT2C receptor gene and human patients with bipolar affective disorder. AB - Two recently described adjacent DNA polymorphisms [(GT)12-18 and (CT)4-5] in the 5'-regulatory region of 5-HT2C receptor gene were analysed in a sample of 88 bipolar patients and 162 controls, all of Spanish origin. Statistical analyses revealed no overall allele or genotype associations with the disease. A haplotype analyses between the (GT)12-18/(CT)4-5 motif and a Cys23Ser variant of the 5-HT2C gene (which had previously been genotyped in the same sample) showed similar distributions between cases and controls. Only a slight increase of s-Ser23 haplotype was found in the subgroup of bipolar women with family history of psychiatric illness (OR=1.24 [95%CI: 1.12-1.38]). PMID- 11502364 TI - Genetic polymorphism in the cathepsin G gene and the risk of Alzheimer's disease. AB - Alzheimer's disease (AD) is a complex disease with the possible involvement of several genes. The APOE*4 allele has been documented to be a major risk factor for sporadic late-onset AD, but it is neither necessary nor sufficient to cause the disease. Cathepsin G, a serine protease found commonly in the azurophillic granules of neutrophils, has been reported to possess some beta-secretase like properties, and thus may be involved in the processing of amyloid precursor protein (APP). Recently, an A-->G polymorphism has been reported in exon 4 of the cathepsin G gene, which changes the codon AAC ((125) Asp) to AGC ((125)Ser). In this study, we have investigated the association of this polymorphism with sporadic late-onset AD. We screened DNA samples from 464 late-onset AD cases and 310 age-matched controls. No significant association was seen between this polymorphism and AD. When the data were stratified by the APOE*4 carrier status, no significant difference was seen either. Our data show no effect of this cathepsin G polymorphism in AD. Characterization of additional polymorphisms in this gene may provide more conclusive answers. PMID- 11502365 TI - Immunisation against East Coast fever by the infection and treatment method: evaluation of the use of ice baths for field delivery and appraisal of an acid formulation of long-acting tetracycline. AB - Immunisation by the infection and treatment method using the Katete strain is currently the most efficient prophylactic technique to control East Coast fever (ECF) in the endemic areas of the Eastern Province of Zambia. The maintenance of the cold chain in liquid nitrogen up to the time of inoculation and the cost of the reference long-acting oxytetracycline (Terramycin LA, Pfizer) are the main drawbacks of the method. The work presented in this paper aims at reducing the cost of immunisation against ECF by using an ice bath for the field delivery and a cheaper long-acting oxytetracycline formulation as chemotherapeutic agent. In experimental conditions, the results from 40 calves immunised after various periods of storage on ice ranging from 4 to 32 h indicate that deferred immunisation performed with a stabilate kept on ice for up to 6h after thawing has an efficiency of 90%. Moreover, sporozoites kept on ice were still surviving 32 h after thawing. In a field trial, 91 calves were inoculated with a stabilate kept for 3.5-5.5 h after thawing and dilution whereas 86 calves were immunised using the standard method. Clinical and parasitological reactions to immunisation were monitored as well as the seroconversion. In the field trial, the deferred immunisation was more efficient than the standard method. The acid formulation of oxytetracycline that was tested was found as suitable as the reference alkaline formulation for the chemotherapeutic control of the Katete strain in ECF immunisation. One indoor trial was carried out on 10 animals and a field trial involved 93 calves. PMID- 11502366 TI - A survey of Theileria sergenti infection, daily weight gain and conception proportions in 85 herds of grazing heifers in Japan. AB - A survey of Theileria sergenti infections, daily weight gain and conception proportion was conducted in 85 herds of grazing heifers in Japan. Basic information and epidemiological data from participating herds were obtained by mailed questionnaires, which were completed by field veterinarians. The average cumulative incidence and proportion of treated animals in the participating herds were 25.7 and 21.1%, respectively. The average daily weight gain and conception proportion were 0.51 kg per day and 56.9%, respectively. The basic information and epidemiological data had a large range and standard deviation, which reflect the wide diversity of the grazing herds in Japan. Herds with heavy tick infestation had significantly higher cumulative incidence and proportion of treated animals, therefore, this factor can be a good estimator to predict the occurrence and loss by theileriosis of the herds. The present questionnaire survey was useful for obtaining information about herds in different regions, and this survey method can be applied to the research of other animal diseases in Japan. PMID- 11502367 TI - Efficacy of combined chemotherapy against gastrointestinal nematodes and Fasciola hepatica in cattle. AB - A controlled trial of the efficacy of several anthelmintic compounds as a combined therapy in the treatment of gastrointestinal nematodes (GIN) and Fasciola hepatica (F. hepatica) in naturally infected cattle was carried out. Twenty crossbred calves, 8-18 months old, were selected for inclusion in the trial based on finding eggs of F. hepatica and GIN in the faeces. They were blocked in four groups of five animals each according to GIN fecal egg counts on day 0. Treatments were sequentially allocated to animals in each block as follows: Group 1 served as non-treated control; Group 2 was treated with netobimin orally at 20 mg/kg; Group 3 received triclabendazole orally at 12 mg/kg and levamisole was applied intramuscularly at 5.5 mg/kg; Group 4 received clorsulon administered subcutaneously (s.c.) at 2 mg/kg and ivermectin s.c. at 200 microg/kg. Six to eight days after treatment the animals were euthanatized in order to collect and identify the parasites. Results showed a reduction of GIN by 87.3, 95.8 and 99.5% in Groups 2, 3 and 4, respectively. The percentage reduction of immature flukes was 0.0, 72.5, and 67.5% and for adult flukes 91.0, 97.5 and 100% for Groups 2, 3 and 4, respectively. Compounds indicated against nematodes showed high efficacy and products directed against F. hepatica acceptably removed adult flukes. However, efficacy against immature stages was generally not satisfactory. PMID- 11502368 TI - Direct anthelmintic effects of condensed tannins towards different gastrointestinal nematodes of sheep: in vitro and in vivo studies. AB - In vitro and in vivo studies were conducted to determine possible direct anthelmintic effects of condensed tannins towards different ovine gastrointestinal nematodes. A larval development/viability assay was used to investigate the effect of a condensed tannin extract (Quebracho) towards larvae of Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus vitrinus. The development to infective larvae and their viability was assessed in all three species and LD 50 values were calculated. The presence of Quebracho extract in the cultures decreased the viability of L3 in all species; the LD 50 were not significantly different for the different species. Forty-eight sheep were allocated to one of eight groups and were infected with a single dose of either 4000 L3 H. contortus (groups 1 and 2) or 5000 L3 T. colubriformis and 5000 L3 Nematodirus battus simultaneously (groups 3-6) or 10,000 L3 of T. circumcincta (groups 7 and 8). From day 28 until day 31 of the experiment, sheep infected with the intestinal species were drenched with Quebracho extract at 4, 8 or 16% w/w of food intake, or remained as undrenched controls; sheep infected with the abomasal species were either drenched with Quebracho extract at 8% w/w of food intake or remained as undrenched controls. All sheep were slaughtered 4 days after the end of the drenching period. Sheep infected with the intestinal species and drenched with 16% w/w Quebracho had lower FEC compared to sheep drenched with 8% w/w (P<0.05), which in turn were lower than in sheep either drenched with 4% Quebracho or which remained undrenched (P<0.05). The lowest intestinal worm burden was recovered from sheep drenched with 8% w/w Quebracho extract (P<0.05). The administration of Quebracho extract at 8% of food intake for 3 days did not affect FEC or worm burdens in sheep infected with the abomasal species compared to controls. PMID- 11502369 TI - Bronchopulmonary helminths of roe deer (Capreolus capreolus) in the northwest of Spain. AB - From 1993 to 1995, 148 hunter killed roe deer (Capreolus capreolus L.) from game reserves in Lugo (northwestern Spain) were examined for lungworm infection. The overall prevalence of infection was high (66.2%). Infections were comprised to two species, Varestrongylus capreoli and Dictyocaulus noerneri with a prevalence of 62.0 and 18.2%, respectively. Significant differences in the prevalence and larval burdens in lungs and faecal samples were observed between different locations. The highest overall prevalence of lungworm infection was observed in the large reserve at Ancares (78%) in comparison to that in smaller reserves (61.7%). Considering that the species recorded in this study are highly host specific, it is unlikely that roe deer represent a significant reservoir of lungworm transmissible to domestic ruminants in northwestern Spain. PMID- 11502370 TI - Stage specific mortality and humoral immune responses during pulse and trickle infestations of the common cattle grub, Hypoderma lineatum (Diptera: Oestridae). AB - Stage specific survival of larvae and pupae of the common cattle grub, Hypoderma lineatum de Villers, was compared in cattle artificially infested with larvae delivered by subcutaneous injection as a single infestation of 100 larvae, in trickle infestations of either two doses of 50 larvae, separated by 15 days, or in four doses of 25 larvae, separated by 9, 6, and 6 days, respectively. The duration of the migratory and 'warble' phases were also compared between treatments. The kinetics of the antibody response was monitored throughout the larval development by ELISA. No difference was noted in the level of antibodies among the three treatments. Mortality of first-instars differed among the infestation groups with mortality in the single pulse infestation group 0.05). Using iodometric technique, in a microtitration tray, the enzyme was fully demonstrated in 10(40%) of the isolates within 30 min, in 22(88%) within 60 min and in all (25 or 100%) within 90 min. The rapid detection of Aeromonas amylase will, no doubt, be of great value in routine diagnostic microbiology. PMID- 11502375 TI - Infectious etiologies in acute exacerbation of COPD. AB - Acute exacerbation (AE) is a frequent episode during the prolonged chronic course of chronic obstructive pulmonary disease (COPD), which entails significant morbidity and mortality. The purpose of this study was to determine the frequency distribution of infectious etiologies in these episodes. Two hundred forty hospitalizations for AECOPD were included in a prospective, purely serologically based study. Paired sera were obtained for each of the hospitalizations and were tested using immunofluorescence or EIA methods to identify 13 different pathogens. Only significant changes in antibody titers were considered diagnostic. The mean age ( +/- SD) of the patients was 66.8 +/- 9.0 years and 179 (84%) were males. In 175 (72.9%) hospitalizations at least one infectious etiology was identified. In 117 (48.8%) hospitalizations at least one of 7 viral etiologies was identified. In 72 (30.0%) hospitalizations at least one of the following atypical bacteria was identified: Legionella spp. in 40 (16.7%), Mycoplasma pneumoniae in 34 (14.2%), and Coxiella burnetii in a single hospitalization. In 58 (24.2%) hospitalizations at least one classic bacterial etiology was found: Streptococcus pneumoniae in 48 (20.0%), Hemophilus influenzae in 10 (4.2%) and Moraxella catarrhalis in 9 (3.8%). More than one etiology was found in 72 (30.0%) hospitalizations. There were no significant differences in the etiologic distribution when the patients were classified by severity of airway obstruction or the clinical type of the exacerbation. We conclude that in most cases of hospitalization due to AECOPD the infectious etiology is viral or atypical bacteria and is classic bacteria in only a minority of cases. More than one etiologic cause can be identified in a third of the cases. The frequency distribution of the etiologies is not associated with the severity of airway obstruction or the clinical type of the exacerbation. The results of our study suggest that atypical bacteria should be covered in antibiotic regimens recommended for AECOPD. This issue should be addressed in future studies. PMID- 11502376 TI - Isolation of Clostridium innocuum from cases of recurrent diarrhea in patients with prior Clostridium difficile associated diarrhea. AB - Clostridium innocuum isolates resistant to vancomycin (MIC values of 16-24 microg/mL) were isolated from three patients with recurrent Clostridium difficile -associated diarrhea (CDAD). We discuss the clinical significance and problems associated with the identification and differentiation of these two clostridial species, which may result in misdiagnosis of patients. PMID- 11502377 TI - Comparison of Light-Cycler PCR, enzyme immunoassay, and tissue culture for detection of herpes simplex virus. AB - A rapid PCR method was developed using the Roche LightCycler technology for detection of Herpes Simplex Virus I and II. The Lightcycler method was compared with tissue culture and direct HSV antigen detection using routine clinical samples. The LightCycler PCR was shown to be more sensitive than isolation by tissue culture (sensitivity of culture versus PCR 78%) and HSV antigen detection (sensitivity of EIA versus PCR 56%). The amplified product may be typed without additional tests. The LightCycler PCR method provides a rapid method for HSV detection that correlates well with established tests. PMID- 11502378 TI - Correlation of MIC methods and tentative interpretive criteria for disk diffusion susceptibility testing using NCCLS methodology for fusidic acid. AB - Three hundred and eight staphylococci and 43 Streptococcus pyogenes were tested by agar dilution, microbroth dilution, E-test, and disk diffusion using 5, 10 and 50 microg disks, as outlined by the NCCLS, in order to correlate the different MIC methods and to establish tentative species specific interpretive zone diameter breakpoints for fusidic acid. MIC results of the three methods were comparable. For Staphylococcus aureus, using MIC breakpoints of < or = 0.5 mg/L for susceptible and > or = 2 mg/L for resistant tentative interpretive zone diameters of > or = 20 mm and > or = 21 mm for susceptible and < or = 17 mm and < or = 18 mm for resistant are suggested for the 5 microg and the 10 microg disk, respectively. The 50 microg disk did not separate susceptible from resistant isolates. For streptococci a uniform MIC distribution of 2-8 mg/L was found. PMID- 11502379 TI - Interactions of colistin and rifampin on multidrug-resistant Acinetobacter baumannii. AB - The increased incidence of nosocomial infections by multidrug-resistant Acinetobacter spp creates demand on the application of some combinations of older antimicrobials on that species. The in vitro activities of colistin and of rifampin and of their interaction were tested on 39 nosocomial isolates of Acinetobacter baumannii. All isolates were resistant to ampicillin/sulbactam, to 3(rd) and 4(th) generation cephalosporins, to amikacin and to ciprofloxacin. MICs were determined by a microdilution technique and interactive studies between 1x or 4x MIC of colistin and rifampin were performed by the time-kill assay. Rifampin was applied at a concentration of 2 microg/mL which is equal to its mean serum level. All isolates were inhibited by colistin and only 15.2% by rifampin. Synergy between 1x MIC of colistin and rifampin was detected in 15.4% of isolates at 6 h of growth and in 51.3% of isolates at 24 h of growth. Synergy between 4x MIC of colistin and rifampin was detected in 15.4% of isolates at 6 h of growth and in 66.7% of isolates at 24 h of growth. It is concluded that colistin is highly active on multidrug-resistant Acinetobacter spp and its activity on A.baumannii is increased in the presence of rifampin, so that their administration might be proposed for nosocomial infections by these isolates. PMID- 11502380 TI - Candida albicans spinal epidural abscess secondary to prosthetic valve endocarditis. AB - A 56-year-old woman, with underlying rheumatic heart disease status post mitral valve replacement, presented with fever, low back pain radiating to right leg, and congestive heart failure. Magnetic resonance imaging detected an L5-S1 spinal epidural abscess. A vegetation on prosthetic mitral valve was found by transesophageal echocardiography. Cultures of epidural aspirate, surgical specimen, and blood all grew Candida albicans. She received surgical drainage of the spinal epidural abscess and i.v. amphotericin B 1 mg/kg/day for eight weeks. Clinical symptoms improved gradually and she was discharged without neurologic sequelae. She remained well and continued to lead an active life two years after discharge. PMID- 11502381 TI - Identification of Arcobacter cryaerophilus isolated from a traffic accident victim with bacteremia by 16S ribosomal RNA gene sequencing. AB - Traditional ways of identifying slow growing bacteria is slow and often difficult. In this study, a small, Gram-negative, facultative anaerobic, slow growing bacillus was isolated from the blood culture of a 7-year old traffic accident victim. The bacterium was non-hemolytic, catalase and oxidase positive. An attempt to use the Vitek system (GNI+) and the API system (20NE) to identify the strain was unsuccessful as the growth controls showed negative results. 16S ribosomal RNA gene sequencing showed that there was 1 base difference between the isolate and Arcobacter cryaerophilus (GenBank Accession no. U25805), 1 base difference between the isolate and A. cryaerophilus (GenBank Accession no. U34387), 10 base differences between the isolate and A. cryaerophilus (GenBank Accession no. L14624), 34 base differences between the isolate and A. butzleri (GenBank Accession no. U34386), 34 base differences between the isolate and A. butzleri (GenBank Accession no. U34387), and 38 base differences between the isolate and A. butzleri (GenBank Accession no. L14626), indicating that the isolate most closely resembled a strain of A. cryaerophilus. Identification of the isolate in our case by conventional methods was difficult, as the absence of a curved morphology has made it confused with other Gram-negative non fermentative bacteria, and the slow growth rate has made it unidentifiable by both the Vitek and API systems. Although the exact source of infection and route of transmission in our case remains elusive, we speculate that the bacteria were transmitted through the respiratory tract while the boy was suffocated in the mud. The present report represents an example of showing the usefulness of 16S rRNA gene sequencing for identification of slow growing bacteria. PMID- 11502382 TI - Epidemiology and frequency of resistance among pathogens causing urinary tract infections in 1,510 hospitalized patients: a report from the SENTRY Antimicrobial Surveillance Program (North America). AB - Bacterial urinary tract infections (UTIs) are an important cause of septicemia resulting in high mortality rates, prolonged hospital stays and increased healthcare costs. Periodic reviews of pathogen frequency and susceptibility patterns impact on appropriate antimicrobial usage, leading to more effective prescribing practices. As part of the SENTRY Antimicrobial Surveillance Program (SENTRY, 1998), participants collected 50 consecutive UTI pathogens from patients hospitalized in 31 medical centers (26 in the United States and five in Canada) and forwarded subcultures to the coordinating center. Thirty-four antimicrobial agents were tested including two investigational compounds (quinupristin/dalfopristin [Q/D], gatifloxacin). The rank order of the 32 species identified during the study was: Escherichia coli (46.9%) > Enterococcus spp. (12.8%) > Klebsiella spp. (11.0%) > Pseudomonas aeruginosa (7.5%) > Proteus mirabilis (5.0%) > coagulase-negative staphylococci (CoNS; 3.4%). This pathogen rank order did not change from 1997 to 1998, but resistance patterns changed. Clonal spread of confirmed extended spectrum beta-lactamase-producing strains was not observed, but co-resistance was elevated for aminoglycosides, tetracyclines, sulfonamides, and fluoroquinolones. P. aeruginosa was most susceptible to amikacin (97.3%) > piperacillin +/- tazobactam (92.0-95.6%) > cefepime = imipenem (91.2%) > ceftazidime (85.8%). Fluoroquinolone resistance was greater in P. aeruginosa (24.8-39.8%) > P. mirabilis (5.3-13.3%) > Enterobacter spp. (6.7-8.9%) > Klebsiella spp. (4.2-7.8%) > E. coli (3.0-3.8%). Only 5% of enterococci were resistant to vancomycin. These results emphasize the need for continued surveillance studies for common infections which establish baseline resistance patterns by geographic areas, and have the potential to detect epidemics or direct local epidemiologic interventions. PMID- 11502385 TI - Genetic schemes and schemata in neurophysiology. AB - Information in nervous systems is often carried by neural ensembles--groups of neurons in transient functional linkage--and written in a code that involves the spatial locations of active neurons or synapses and the times at which activity occurs. Even in favorable neuroanatomical circumstances, studying neural ensemble function presents a serious experimental challenge. One recent strategy to overcome this challenge relies on protein-based sensors that provide direct optical images of neural activity, and on protein-based effectors that interfere with it. Because these molecules are encodable in DNA, they can be introduced into intact animals by genetic manipulation, and their expression pattern can be tailored to include--exclusively and at the same time comprehensively--the neurons of interest. Circumscribed populations of neurons can thus be studied in virtual isolation at defined stages of intact neural pathways. PMID- 11502383 TI - Antibiotic usage in primary care units in Taiwan after the institution of national health insurance. AB - To understand antibiotic usage in primary care units in Taiwan after the institution of national health insurance, we collected all prescriptions for one week in March each year from 1996 to 1999 from out-patient clinics of randomly sampled public health stations located in various parts of Taiwan. We sampled 114 health stations with 40,891 patient-visits and 68,386 diagnoses made in 1996; 154 health stations were sampled with 53,992 patient-visits and 99,466 diagnoses in 1997; 166 health stations were sampled with 49,112 patient-visits and 96,161 diagnoses in 1998; and 162 health stations were sampled with 46,976 patient visits and 93,641 diagnoses in 1999. The percentages of total patient-visits that resulted in antibiotic treatment were 14.2%, 12.5%, 14.1% and 13.0% in the four time periods, respectively. Among patient-visits, patient group under the age of 11 received the highest percentage of antibiotic treatment. Common cold was the most frequent diagnosis for which antibiotic was prescribed during the four time periods, accounting for 32.3% of total antibiotic prescriptions. Among patients with the diagnosis of common cold, 31.3% received antibiotic treatment, and the highest figure was among pediatric patients (<16 year of age). Penicillins, cephalosporins and macrolides were the most commonly prescribed classes of antibiotics. They accounted for 35.4%, 26.5% and 21.6% of all prescribed antibiotics in these four study periods. From this study, it was found that, after institution of the national health insurance system, antibiotics are still very commonly used in primary care units in Taiwan and still seem to be overused. PMID- 11502386 TI - Dendritic processing. AB - Dendritic processing multiplies the computational power of a single neuron by enabling the processing of inputs in a spatio-temporally differentiated manner. Recently, the development of new and refined optical, electrophysiological and molecular-biological techniques has led to new insights into dendritic function and revealed an astonishing plethora of computational mechanisms. PMID- 11502387 TI - Neurodynamics: nonlinear dynamics and neurobiology. AB - The use of methods from contemporary nonlinear dynamics in studying neurobiology has been rather limited.Yet, nonlinear dynamics has become a practical tool for analyzing data and verifying models. This has led to productive coupling of nonlinear dynamics with experiments in neurobiology in which the neural circuits are forced with constant stimuli, with slowly varying stimuli, with periodic stimuli, and with more complex information-bearing stimuli. Analysis of these more complex stimuli of neural circuits goes to the heart of how one is to understand the encoding and transmission of information by nervous systems. PMID- 11502388 TI - Neuronal diversity in the retina. AB - The listing of cell types present in the retina is nearing completion, the first time this can be said for any significantly complex sample of the central nervous system. Mammalian retinas contain approximately 55 separate neuronal types. The functions of 22 of them are known or can be strongly inferred. For these 22, in every instance, a cell defined as a 'type' by structural criteria carries out a distinct and individual physiological function. Electrophysiological experiments continue to reveal new features of the retina's handling of information, and there is every reason to believe that the remaining 33 types of cell will also have distinct physiological functions. Further subtleties clearly exist in both peripheral and central visual coding. PMID- 11502389 TI - How do visual neurons respond in the real world? AB - Natural visual stimuli have been reported to evoke neural responses with brief, intense firing events. This has important implications for how visual information is encoded. A recent experiment measured the responses of visual neurons in the natural environment, circumventing many limitations of laboratory simulations while still retaining tight experimental control. Under these conditions, neurons fired intensely at precise, discrete times, validating that this firing regime is relevant to natural vision. PMID- 11502390 TI - Analysis of sensory coding with complex stimuli. AB - Most high-level sensory neurons have complex, nonlinear response properties; a comprehensive characterization of these properties remains a formidable challenge. Recent studies using complex sensory stimuli combined with linear and nonlinear analyses have provided new insights into the neuronal response properties in various sensory circuits. PMID- 11502391 TI - Can you still see the cochlea for the molecules? AB - It is now established that the mammalian cochlea uses active amplification of incoming sound to achieve sensitivity. Cellular details are emerging slowly. Recent studies of sensory hair cells have highlighted the possible molecular bases for amplification and the components of sensitivity regulation within the cochlea: a synthesis is likely to depend on effective and physiologically informed modelling. PMID- 11502392 TI - The roles and functions of cutaneous mechanoreceptors. AB - Combined psychophysical and neurophysiological research has resulted in a relatively complete picture of the neural mechanisms of tactile perception. The results support the idea that each of the four mechanoreceptive afferent systems innervating the hand serves a distinctly different perceptual function, and that tactile perception can be understood as the sum of these functions. Furthermore, the receptors in each of those systems seem to be specialized for their assigned perceptual function. PMID- 11502393 TI - Magnetite-based magnetoreception. AB - Orientation, navigation, and homing are critical traits expressed by organisms ranging from bacteria through higher vertebrates. Sensory systems that aid such behavior have provided key selective advantages to these groups over the past 4 billion years, and are highly evolved; magnetoreception is no exception. Across many species and groups of organisms, compelling evidence exists that the physical basis of this response is tiny crystals of single-domain magnetite (Fe3O4). It is the opinion of the authors that all magnetic field sensitivity in living organisms, including elasmobranch fishes, is the result of a highly evolved, finely-tuned sensory system based on single-domain, ferromagnetic crystals. PMID- 11502394 TI - Recent dynamics in olfactory population coding. AB - Recent studies of the olfactory bulb (vertebrates) and antennal lobe (insects) have elucidated how odor information is represented by the identity, synchronization, slow temporal dynamics and position of active neurons in an ensemble. Odor representations are dynamically reorganized both during the course of a stimulus and with experience. These results provide new insights into the logic of odor representations and dynamical network computations. PMID- 11502395 TI - Energy as a constraint on the coding and processing of sensory information. AB - Neurons use significant amounts of energy to generate signals. Recent studies of retina and brain connect this energy usage to the ability to transmit information. The identification of energy-efficient neural circuits and codes suggests new ways of understanding the function, design and evolution of nervous systems. PMID- 11502396 TI - Memory-based expectations in electrosensory systems. AB - Adaptive processing of electrosensory information occurs in the cerebellum-like structures of three distinct groups of fish. Associations within each of these structures result in the generation of negative images of predictable features of the sensory inflow. Addition of these negative images to the actual inflow removes the predictable features, allowing the unpredictable, information-rich sensory signals to stand out. Evidence from all three groups of fish indicates that the negative images are mediated by plasticity at parallel fiber synapses. PMID- 11502397 TI - Processing in layer 4 of the neocortical circuit: new insights from visual and somatosensory cortex. AB - Recent experimental and theoretical results in cat primary visual cortex and in the whisker-barrel fields of rodent primary somatosensory cortex suggest common organizing principles for layer 4, the primary recipient of sensory input from the thalamus. Response tuning of layer 4 cells is largely determined by a local interplay of feed-forward excitation (directly from the thalamus) and inhibition (from layer 4 inhibitory interneurons driven by the thalamus). Feed-forward inhibition dominates excitation, inherits its tuning from the thalamic input, and sharpens the tuning of excitatory cells. Recurrent excitation enhances responses to effective stimuli. PMID- 11502398 TI - The organization of sensory cortex. AB - Recent studies of primary visual cortex (V1) redefine layers 3 and 4 of V1 in monkeys and show that monkeys, apes and humans have different laminar specializations. Projections from V1 define a smaller, but complete, third visual area, and a dorsomedial area. The middle temporal visual area has two types of motion-sensitive modules with inputs from cytochrome oxidase columns in V1. Second-level somatosensory areas have been described in humans, and a second level auditory area is shown to respond to somatosensory stimuli. PMID- 11502399 TI - Sensory modalities are not separate modalities: plasticity and interactions. AB - Historically, perception has been viewed as a modular function, with the different sensory modalities operating independently of each other. Recent behavioral and brain imaging studies challenge this view, by suggesting that cross-modal interactions are the rule and not the exception in perception, and that the cortical pathways previously thought to be sensory-specific are modulated by signals from other modalities. PMID- 11502400 TI - Learning to communicate. AB - Of the few animal groups that learn their vocalizations, songbirds are uniquely amenable to molecular, physiological, and behavioral analyses of the neural features responsible for vocal learning. In order to communicate effectively as an adult, a young songbird recognizes and memorizes a model of his species specific song during a developmentally critical period called sensory acquisition. Factors are now emerging that contribute to the length and strength of this learning phase. In a second critical period, known as sensorimotor learning, the young bird uses auditory feedback to perfect his motor performance, creating a match to the memorized model. New studies show that motor matching can persist beyond sensorimotor learning, and thus a role for the acquired model might also persist into adulthood. Fascinating in their own right, songbirds also provide optimism that mature brains have recourse to plasticity. PMID- 11502401 TI - The importance of the mechanical circulatory support database: a plea for commitment. PMID- 11502402 TI - The Registry of the International Society for Heart and Lung Transplantation: eighteenth Official Report-2001. PMID- 11502403 TI - Transplantation tolerance induced by mixed chimerism. AB - Although short- and long-term results after organ transplantation have improved considerably in recent years, morbidity and mortality rates in graft recipients remain high. The induction of lifelong donor-specific tolerance would dramatically improve outcome after organ transplantation. Although many tolerance protocols have been successful in rodent studies, most of these approaches have failed when attempted in large animals or humans. Robust tolerance, in contrast, has been demonstrated with mixed chimerism regimens not only in rodents but also in large animal models, including non-human primates. Furthermore, mixed chimerism protocols have been developed that would be feasible in cadaveric, and thus in thoracic, transplantation. The induction of mixed hematopoietic chimerism is therefore an attractive experimental approach for development of clinical tolerance protocols. One of the obstacles to widespread clinical application of this concept is the remaining toxicity of the host conditioning. Recent advances, however, have led to substantially milder protocols that could become clinically acceptable in the foreseeable future. This article provides a short overview of the basic mechanisms by which immunologic tolerance may be induced, describes the concept of mixed chimerism as a promising approach for clinical tolerance induction, and reviews recent progress in developing clinically feasible mixed chimerism protocols. PMID- 11502404 TI - Exaggerated chronotropic and energetic response to dobutamine after orthotopic cardiac transplantation. AB - BACKGROUND: After heart transplantation, the transplanted denervated heart displays both an exaggerated chronotropic and an exaggerated inotropic response to circulating catecholamines. This study assessed whether denervated transplanted hearts also display an exaggerated energetic response when challenged with dobutamine. METHODS AND RESULTS: A total of 18 heart transplant recipients and 14 normal volunteers underwent measurements of myocardial oxygen consumption (MVO2), external work (EW), and pressure-volume area (PVA), at rest and during infusion of dobutamine. At rest, calculated myocardial (PVA/MVO2) and mechanical (EW/MVO2) efficiencies were similar among transplant recipients and normal volunteers. During low-dose dobutamine infusion (8 microg/kg/min), transplant recipients exhibited a larger increase in heart rate (to 126 +/- 14 vs 87 +/- 26 beats/min, p < 0.001) and MVO2 (to 269 +/- 43 vs 233 +/- 19 J/min/100g, p < 0.05) and a smaller increase in EW (64 +/- 18 vs 72 +/- 13 J/min/100g, p < 0.05) and PVA (70 +/- 16 vs 81 +/- 13 J/min/100g, p < 0.05) than did normal volunteers. As a result, both myocardial (26 +/- 4 vs 35 +/- 4%, p < 0.05) and mechanical (23 +/- 4 vs 30 +/- 4%, p < 0.001) efficiencies were lower during dobutamine infusion in transplant recipients than in normal volunteers. During the infusion of a higher dose of dobutamine (19 microg/kg/min), the chronotropic and inotropic responses of heart transplant recipients were even more exaggerated. The fall in myocardial efficiency induced by dobutamine correlated with the increase in heart rate (r = -0.58) and could be reproduced in normal volunteers by coadministration of atropine. CONCLUSIONS: Transplant recipients exhibit a larger fall in contractile efficiency and a larger oxygen-wasting effect during dobutamine infusion than do normal volunteers. Because normal volunteers pre-medicated with atropine presented with a similar increase in heart rate and a similar fall in efficiency, the exaggerated energetic response of transplanted hearts to dobutamine likely resulted from the same mechanisms as their chronotropic supersensitivity, i.e., the loss of inhibitory parasympathetic innervation. PMID- 11502405 TI - Pulmonary artery systolic pressures estimated by echocardiogram vs cardiac catheterization in patients awaiting lung transplantation. AB - BACKGROUND: At many lung transplant centers, right heart catheterization and transthoracic echocardiogram are part of the routine pre-transplant evaluation to measure pulmonary pressures. Because decisions regarding single vs bilateral lung transplant procedures and the need for cardiopulmonary bypass are often made based on pulmonary artery systolic pressures, we sought to examine the relationship between estimated and measured pulmonary artery systolic pressures using echocardiogram and catheterization, respectively. METHODS: We retrospectively reviewed all patients in our program who had measured pulmonary hypertension (n = 57). Patients with both echocardiogram-estimated and catheterization-measured pulmonary artery systolic pressures performed within 2 weeks of each other were included (n = 19). We analyzed results for correlation and linear regression in the entire group and in the patients with primary pulmonary hypertension (n = 8) and pulmonary fibrosis (n = 8). RESULTS: In patients with primary pulmonary hypertension, pulmonary artery systolic pressure was 94 +/- 27 and 95 +/- 15 mm Hg by echocardiogram and catheterization, respectively, with r(2) = 0.11; in patients with pulmonary fibrosis, 57 +/- 23 and 58 +/- 12 mm Hg with r(2) = 0.22; and in the whole group, 76 +/- 29 and 75 +/ 23 mm Hg with r(2) = 0.50. Thirty-two additional patients had mean pulmonary artery systolic pressure = 48 +/- 16 mm Hg by catheterization but either had no evidence of tricuspid regurgitation by echocardiogram (n = 22) or the pulmonary artery systolic pressure could not be measured (n = 10). CONCLUSIONS: In patients with pulmonary hypertension awaiting transplant, pulmonary artery systolic pressures estimated by echocardiogram correspond but do not serve as an accurate predictive model of pulmonary artery systolic pressures measured by catheterization. Technical limitations of the echocardiogram in this patient population often preclude estimating pulmonary artery systolic pressure. PMID- 11502406 TI - Iron overload and nitric oxide-derived oxidative stress following lung transplantation. AB - BACKGROUND: Reactive oxygen species (ROS) may contribute to airway injury and the development of the bronchiolitis obliterans syndrome (BOS) following lung transplantation (LT). Chemically active iron released from ferritin stores and nitric oxide (NO)-derived radicals may add to the oxidative burden. METHODS: We determined the concentrations of ferritin and the aqueous NO derivative nitrite (NO2(-)) within bronchoalveolar lavage fluid (BALF) of 14 stable LT recipients (ST) and 7 subjects with BOS and 21 normal controls. We also assessed the relationship between BALF ferritin and hemosiderin-laden macrophages (HLMs) using a hemosiderin score (HS) and determined BALF albumin concentration as a marker of microvascular leakage. RESULTS: BALF ferritin concentrations and HSs were significantly elevated in LT recipients overall compared with normal controls (p < 0.05). BALF NO2(-) levels were elevated in BOS subjects and STs compared with normal controls (p = 0.002 and p = 0.09, respectively), but there was no difference between transplant groups. BALF albumin concentrations were elevated in BOS patients compared with normal controls (p = 0.02) and ST (p = 0.05), but there was no difference between STs and controls. There was a significant relationship between BALF ferritin concentration and HS in LT recipients overall (r(s) = 0.7, p < 0.001). In BOS subjects, but not ST, BALF ferritin was significantly related to BALF albumin (r(s) = 0.8, p = 0.05) and there was a weak relationship with NO2(-) concentration (r(s) = 0.6, p = 0.1). BALF NO2(-) was strongly related to BALF % neutrophils in BOS subjects (r(s) = 0.9, p < 0.01), but there was no such relationship in STs. CONCLUSIONS: Our findings suggest that the allograft could be subject to significant iron-generated oxidative stress, which may be exacerbated by NO and neutrophil-derived ROS, particularly in BOS. Microvascular leakage may be a feature of established chronic rejection, which potentiates the iron overload and contributes to further airway damage and remodeling. PMID- 11502407 TI - Preliminary result of an algorithm to select proper ventricular assist devices for high-risk patients with extracorporeal membrane oxygenation support. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) is not suitable for long term support because of its high incidence of complications. Conversion from ECMO to ventricular assist device (VAD) is reasonable, and we have developed a simple algorithm for selecting proper VADs for these ECMO-supported patients. METHODS: We converted 12 patients who were receiving ECMO support to VAD for bridge to transplantation. Group I (n = 6) was converted directly from ECMO to VAD. Group II (n = 6) underwent stage conversion. We added left atrial drainage to ECMO because of lung edema or marked left heart distension. We discontinued drainage after recovery of right heart function. Group II had more unfavorable risk factors for VAD before ECMO. RESULTS: Three patients (50%) in Group I received biventricular VADs. The other 3 patients were converted to left ventricular assist device (LVAD), but only 1 (16.7%) experienced successful conversion. We successfully converted 5 patients (83.3%) in Group II to LVAD without right VAD, and 4 of them could be weaned from the ventilator. The multiple-organ dysfunction score gradually improved in Group II despite additional surgery. Two patients in each group received heart transplantation and survived long term. CONCLUSION: Using a conversion protocol provides a good guideline for making decisions. According to the protocol, right heart and pulmonary function can be clearly assured before shifting to LVAD in these critical ECMO-supported patients. PMID- 11502408 TI - The topography of intimal thickening and associated remodeling pattern of early transplant coronary disease: influence of pre-existent donor atherosclerosis. AB - BACKGROUND: With native coronary disease, intimal plaque initially accumulates at focal areas in the artery, often accompanied by compensatory vessel enlargement. With transplant coronary disease, the topography of intimal thickening and associated remodeling pattern are less studied. METHODS: We studied 72 prospectively recruited transplant patients with serial intravascular ultrasound using 4.3F catheters at baseline and at 1-year follow up. We considered 175 ultrasound-recorded segments (mean, 2.4 +/- 1.1 segments per patient) exactly matched on the serial studies by both angiographic criteria and ultrasound criteria, using arterial and venous branch points, pericardium, and sinuses as anatomic landmarks. RESULTS: Eighty-eight segments had no donor disease, and 87 had donor disease (80 eccentric and 7 concentric intimal thickening). Progressive intimal thickening occurred in 48 segments without (55%) and 43 segments with donor disease (48%, p = NS). Thickening from segments without donor disease was mainly eccentric (81%). Thickening from segments with donor eccentric plaque was also mainly eccentric (67%, p = NS compared with segments without donor disease), with further thickening superimposed on the original plaque. Concentric intimal thickening was uncommon. Of the 58 patients who had >1 segment matched, intimal changes were discordant in 34 (59%), with progression in some and lack of progression in other segments. Total vessel area change correlated with intimal area change (r = 0.37 with a slope of 0.79, p < 0.001), including segments with (r = 0.39; slope, 0.69) and segments without (r = 0.37; slope, 1.16) donor disease. CONCLUSION: The intimal thickening of early transplant coronary disease is mainly eccentric and often discordant within each individual patient. Donor eccentric plaque often serves as a nidus for further intimal growth. The topography of intimal thickening in transplant coronary disease resembles that of native coronary disease, but the presence of a pre-existent donor plaque may impede compensatory remodeling as further intimal thickening occurs after transplantation. PMID- 11502409 TI - Hepatitis B virus infection in heart transplant recipients in a hepatitis B endemic area. AB - BACKGROUND: Hepatitis B virus (HBV) infection is hyperendemic in Taiwan. It is almost impossible for us to reject organ donors or recipients with positive serum hepatitis B surface antigen (HBsAg). We report our experience with HBV infection in heart transplant recipients with particular attention to outcome of recipients who were HBsAg+ or who had received donor hearts from HBsAg+ donors. METHODS: We performed a retrospective review of medical records. RESULTS: In the study, we included 101 heart recipients with post-transplant survival of more than 6 months. According to pre-transplant HBV serology markers, we divided patients into 4 groups. Group 1 (n = 8) had been HBsAg+ at the time of heart transplantation. Of these, 6 patients had HBV reactivation in the post-transplant follow-up and needed lamivudine treatment. Complete response was achieved in all 6 patients; however, HBV recurrence occurred in 1 patient after 8 months of lamivudine treatment. The recurrence remained under partial control. Group 2 (n = 16) was HBV naive at the time of heart transplantation. Of these, 2 received HBsAg+ donor hearts under perioperative hepatitis B immunoglobulin prophylaxis. HBV infection was successfully prevented in 1 patient, but the other contracted HBV hepatitis, which was successfully treated with lamivudine. In Group 2, 10 patients received donor hearts from anti-HBs+ donors, and none contracted HBV hepatitis after transplantation. Group 3 (n = 55) had protective anti-HBs antibody at the time of heart transplantation either from previous HBV vaccination (n = 10) or from natural HGB infection (n = 45). HBsAg+ donor hearts were transplanted into 2 patients with anti-HBs from previous HBV vaccination, and into 8 patients with anti-HBs form natural HBV infection. However, none of these 10 patients who received HBsAg+ donor hearts had HBV hepatitis after transplantation. Group 4 (n = 22) was HBs-, anti-HBs-, and anti-HBc+ at the time of heart transplantation. Of these, 7 patients received HBsAg+ donor hearts. Six patients experienced no HBV hepatitis after heart transplantation, and serum HBV DNA by polymerase chain reaction (PCR) at the time of heart transplantation was negative in all 6 patients. One patient had HBV hepatitis after transplantation, and serum HBV DNA by PCR at the time of heart transplantation also was positive. CONCLUSION: HBV reactivation after the heart transplantation was common but usually well controlled with lamivudine treatment. Therefore, HBV carrier status should not contraindicate heart transplantation. HBsAg+ donor hearts were safely transplanted into anti-HBs+ recipients; therefore, HBsAg+ itself was not a contraindication to heart donation. Patients with HBsAg-, anti-HBs-, anti-HBc+, and negative HBV DNA in the serum by PCR could be protected from HBV infection from HBsAg+ donor hearts. However, patients with HBsAg-, anti-HBs-, anti-HBc+, and positive HBV DNA in the serum by PCR should be recognized as HBV carriers and closely followed for potential HBV flare-up after heart transplantation. PMID- 11502410 TI - Nitroglycerin alters alveolar type II cell ultrastructure after ischemia and reperfusion. AB - BACKGROUND: Although administration of nitric oxide (NO) has been suggested to reduce pulmonary reimplantation response, concerns remain about cytotoxic side effects. METHODS: Using light and electron microscopy, we examined the effects of the NO donor nitroglycerin (NTG) (0.1 mg/ml) as a supplement to the preservation solution Celsior on the structural integrity of rat lungs after extracorporeal ischemia (4 hours at 10 degrees C) and reperfusion (50 minutes) (IR). We performed evaluation in comparison with Celsior alone after IR using either standard antegrade perfusion through the pulmonary artery or retrograde perfusion through the left atrium as an alternative way to improve the preservation quality. Untreated, non-ischemic lungs served as controls (n = 5 per group). We recorded respiratory and hemodynamic parameters during reperfusion. Tissue collection using systematic uniform random sampling was representative for the whole organ and allowed stereologic quantification of structures. RESULTS: After IR, histochemistry revealed no breaks in the alveolo-capillary barrier and we detected no alveolar flooding. Edema formed in the peribronchovascular cuffs, of which the volume fraction was increased (p =.008). Vasoconstriction of the smaller arteries accompanied antegrade flush, which occurred neither after administration of NTG nor after retrograde flush, as shown by immunostaining for alpha-smooth muscle actin. Treatment with NTG was associated with focal disintegration of Type II cells, which displayed edematous swelling of distinct cell compartments and lysis of mitochondria and cells. Nitroglycerin prevented alveolar collapse, which was increased in the other IR groups (p = 0.013). We observed alterations in intra-alveolar surfactant components. CONCLUSION: These findings indicate pathologic effects of NTG treatment on alveolar epithelial integrity. Therefore, we suggest further critical evaluation of NTG/NO for therapeutic use in lung transplantation. PMID- 11502411 TI - Effect of a neutrophil elastase inhibitor (ONO-5046 Na) on ischemia/reperfusion injury using the left-sided heterotopic canine heart transplantation model. AB - BACKGROUND: Ischemia/reperfusion injury is a major cause of transplanted heart dysfunction. Several reports have demonstrated that polymorphonuclear neutrophil (PMN) elastase derived from the activated neutrophils might play an important role in this injury. Herein, we investigated the protective effects of PMN elastase inhibitor (ONO-5046 Na) on ischemia/reperfusion injury using a left sided canine heterotopic heart transplantation model. METHODS: We used 10 pairs of adult beagle dogs. The donor heart was transplanted heterotopically into the left thoracic cavity of the recipient without cardiopulmonary bypass. A bolus of ONO-5046 Na (10 mg/kg) was introduced intravenously to 5 recipients (group II) at 15 minutes before reperfusion and was followed by continuous infusion (10 mg/kg per hour) for 180 minutes. Five dogs (group I) did not receive ONO-5046 Na and thus served as a control. After reperfusion, we evaluated transplanted heart function and obtained blood samples from the coronary sinus over a 360-minute period. RESULTS: E(max) and pre-load recruitable stroke work in group II showed significantly better recovery than group I. Blood levels of PMN elastase, creatine kinase MB, lactate and inflammatory cytokines (tumor necrosis factor alpha, interleukin-6, interleukin-8) were significantly lower in group II. Depletion of myocardial concentration of adenosine triphosphate at 120 minutes after reperfusion and myocardial water content was significantly lower in group II. CONCLUSIONS: ONO-5046 Na, which inhibits PMN elastase, could reduce ischemia/reperfusion injury in heart transplantation. These results indicate that clinical application of ONO-5046 Na should be considered. PMID- 11502412 TI - A limited sampling strategy for cyclosporine area under the curve monitoring in lung transplant recipients. AB - We developed a limited sampling strategy (LSS) for predicting cyclosporine (Neoral) area under the curve from concentration-time data obtained specifically from lung transplant recipients. The optimal and most clinically convenient LSS for lung transplant recipients, based on patient wait time, number of blood samples required, percent prediction error, and assessment of predictive performance is one that requires 2 blood samples collected at 1 and 3 hours post dose: AUC = 1.75 x C(1) + 4.91 x C(3) + 185.62. PMID- 11502413 TI - Intracranial bleed during bridge to transplant may not preclude a successful result. AB - We report the case of a 29-year-old man who suffered sub-arachnoid bleeding while stabilized on a biventricular assist device as a bridge to cardiac transplantation. We adjusted his anti-coagulation therapy to control the bleeding and to concurrently minimize thrombosis while on support. He underwent 2 craniotomy operations to evacuate sub-arachnoid hematomas, and he underwent a subsequent operation to debride and close the dura. Eighteen days later, he underwent successful orthotopic heart transplant and was discharged to home 3 weeks post-transplant. PMID- 11502414 TI - Isolated non-compaction of the left ventricle: a rare indication for transplantation. AB - This report describes the diagnostic difficulty encountered in a young female patient presenting with neurologic symptoms, atrial fibrillation and severe left ventricular systolic dysfunction, eventually leading to cardiac transplantation. The scrutiny used in the evaluation of the particular aspect of the left ventricle, and the integration of the information obtained from echocardiography, angiography and magnetic resonance imaging, led to the diagnosis of a rare and mostly unknown cause of cardiac failure. The correct identification of this entity is mandatory because enhanced risk of thromboembolism and malignant arrhythmia should be anticipated. A review of the literature revealed only 6 patients in whom isolated non-compaction of the left ventricle was treated by heart transplantation. PMID- 11502415 TI - Bronchogenic carcinoma in lung transplant recipients. AB - Although lung transplant recipients have a higher prevalence of non-melanoma skin cancers and lymphoma than the general population, the same has not been noted for bronchogenic carcinoma. If an increased prevalence of bronchogenic carcinoma exists, contributing factors may include the high rate of previous tobacco use in this population and/or the chronic immunosuppression used to prevent allograft rejection. With time, the incidence of bronchogenic carcinoma in the lung transplant population is likely to parallel the increasing longevity and number of transplanted individuals. We describe 2 cases of bronchogenic carcinoma in lung transplant recipients that demonstrate the morbidity associated with the discovery or development of bronchogenic carcinoma in this population. PMID- 11502416 TI - Fatal visceral leishmaniasis in a heart transplant recipient. PMID- 11502417 TI - First successful Italian clinical experience with DeBakey VAD. AB - The growing number of patients waiting for heart transplantation more than tripled between 1989 and 1998. Various non-pulsatile mechanical circulatory support devices have been developed as bridge to heart transplantation in recent years. We report the first successful Italian clinical experience with an axial flow pump, DeBakey VAD, in a patient supported as bridge to transplantation for 55 days. PMID- 11502418 TI - Revascularization of the occluded right coronary artery during left ventricular assist device implantation. AB - A 62-year-old man with end-stage ischemic cardiomyopathy and left ventricular function of 20% was evaluated for heart transplantation. Cardiac catheterization revealed proximal occlusion of the dominant right coronary artery (RCA) with collateral blood flow and significant stenosis in the distal part, but no significant re-occlusions of the stented left coronary artery and no significant stenosis of the left circumflex artery. When the patient became catecholamine dependent, Novacor left ventricular assist device (LVAD) implantation, as a bridge to transplantation, was considered and the patient operated upon. To avoid ischemic right heart failure after LVAD implantation, a concomitant re vascularization of the distal RCA was performed. The post-operative course was uneventful. Five weeks later, a control angiogram showed the patent bypass graft. The distal stenosis of the RCA was treated successfully with dilation and stent implantation. The patient is presently in stable condition on LVAD and awaits transplantation as an outpatient. PMID- 11502419 TI - Fast three-dimensional sodium imaging of human brain. AB - A three-dimensional sodium imaging technique with a minimum echo time of 0.9 ms is described in a 2.0 Tesla whole-body system. The relaxation behaviour in vivo of sodium was analysed: a fast T(2)(*) relaxation component between 1.2 and 1.6 ms and a slow T(2)(*) relaxation component between 7.1 ms and 8.4 ms were quantified in brain tissue of three volunteers. Three-dimensional sodium images of the human brain were acquired in 8.5 min with a resolution of 4.7 x 4.7 x 10 mm (0.2 cc voxel size) and a signal-to-noise ratio of 20 in brain tissue and 30 in cerebrospinal fluid. PMID- 11502421 TI - Silent BOLD imaging. AB - Pulsed magnetic field gradients in magnetic resonance imaging produce high levels of acoustic noise. In functional magnetic resonance imaging, acoustic scanner noise causes unwanted masking effects. Recently, we proposed a method to perform magnetic resonance imaging experiments undisturbed by acoustic scanner noise by utilizing the property of standard gradient coils to poorly submit acoustic noise in the low frequency range. The silent gradient scheme is now incorporated into a standard T(2)*-weighted sequence. Additionally, simultaneous multi-slice excitation (SIMEX) pulses were implemented to improve the intrinsic low volume coverage of the silent sequence. The proposed silent SIMEX technique was tested and compared with a standard noisy technique using auditory and visual stimulation paradigms. The scanner noise during the silent experiments could be reduced below the range of the ambient noise of the magnet room. This feasibility study shows a trend of decreased activated areas in the noisy experiment for both, the visual and auditory paradigm. PMID- 11502420 TI - (31)P-MR Spectroscopy for the evaluation of energy metabolism in intact residual myocardium after acute myocardial infarction in humans. AB - OBJECTIVE: Experimental studies have demonstrated that acute myocardial infarction (MI) alters energy metabolism even in non-infarcted adjacent tissue. In patients with subacute MI, the influence of the regional ischemic insult on energy metabolism of intact septal myocardium was analyzed using 31P-Magnetic resonance spectroscopy (MRS). PATIENTS AND METHODS: In eight patients with wall motion abnormalities in the anterior wall 31P-spectra were obtained from non infarcted adjacent septal myocardium, as well as infarcted anterior myocardium (voxel size 25 ccm each) 29+/-8 days after MI using a 3D-CSI technique. Additionally, cardiac function was analyzed using breath-hold cine MRI. MRI was repeated 6 months after revascularization to assess viability of infarcted segments. Eight age-matched healthy volunteers served as control group. RESULTS: According to follow-up MRI 4/8 patients showed regional wall motion recovery. Here, PCr/ATP-ratios were not significantly reduced in intact septal myocardium as well as infarcted anterior myocardium compared to healthy volunteers (1.28+/ 0.10 and 1.14+/-0.09 vs. 1.45+/-0.29). No recovery of regional function was detected in 4/8 patients with-therefore-non-viable anterior myocardium. PCr/ATP ratios were significantly reduced in intact and infarcted myocardium compared with healthy volunteers as well as to patients with wall motion recovery (0.77+/ 0.17 and 0.49+/-0.23; P<0.05). DISCUSSION: These preliminary results indicate that energy metabolism is reduced in patients with persisting wall motion abnormalities after myocardial infarction and revascularization in ischemically injured as well as in adjacent non-injured myocardium. PMID- 11502422 TI - Echo planar magnetic resonance imaging of anisotropic diffusion in asparagus stems. AB - MR diffusion-weighted imaging (DWI) uses the signal loss associated with the random thermal motion of water molecules in the presence of magnetic field gradients to derive a number of parameters that reflect the translational mobility of the water molecules in tissues. In highly organized but asymmetric structures, this mobility may be affected by the obstacles present and this in a direction-dependent way. Important examples of this are white brain matter and the stem of certain plants, both containing fibrous components where diffusion of water molecules across fibers is much more restricted than along the fibers. Diffusion that exhibits such directional dependence is said to be anisotropic, and diffusion tensor magnetic resonance imaging allows localized characterization of this behavior. Interpretation of the information obtained in terms of the underlying tissue structure is often hampered by the complexity of factors that can produce the observed behavior. A phantom that exhibits well-defined anisotropic diffusion and yields sufficient signal can help the experimental testing of the relevant methods and models. In this paper, we have used a phantom consisting of asparagus stems as a test object for assessing the value of the acquisition and postprocessing techniques commonly used in the clinic for this kind of investigation. Because of its strongly fibrous and cylindrically symmetric morphology, exhibiting a well-defined sub-classification of cells on the basis of size and shape, asparagus allows a relatively simple interpretation of the results obtained in the diffusion experiments. Our experiments show that the known structural information about the main cell types encountered correlates well with the behavior patterns of the diffusion parameters. PMID- 11502423 TI - Slice acquisition order and blood oxygenation level dependent frequency content: an event-related functional magnetic resonance imaging study. AB - Many event-related functional magnetic resonance imaging paradigms performed so far have been designed to study a limited part of the brain with high temporal resolution. However, event-related paradigms can be exploratory, therefore requiring whole brain scans and so repetition times (TR) of several seconds. For these large TR values, the slice acquisition order may have an important effect on the detection of event-related activation. Indeed, when the scanning is interleaved, the temporal delay between the acquisition of two contiguous slices can reach a few seconds. During this time, the subject is likely to move, and the haemodynamic response will vary significantly. In this case, the interpolation applied between contiguous slices for motion correction induces a temporal smoothing between voxels that are spatially close but temporally sampled a few seconds apart. This should modify the frequency structure of the response and may impair the detection of short events. We, therefore tested the effect of three acquisition schemes (sequential, sequential with gap and interleaved, INT) at two repetition times (TR=3 and 6 s on six and seven subjects, respectively) on activation detection and frequency content in a visual motion event-related paradigm. Unexpectedly, for large TR (6 s), results were found in favour of the INT acquisition scheme (P<0.05). For smaller TR, no strong bias could be found. Generally, intra-subject variability (across acquisition schemes) is found to be much smaller than inter-subject variability, confirming the importance of multi subjects analyses. Our study also shows that important physiological information is carried by high frequency components that should not be filtered out. PMID- 11502424 TI - Measurements of left ventricular dimensions using real-time acquisition in cardiac magnetic resonance imaging: comparison with conventional gradient echo imaging. AB - This study investigates the use of real-time acquisition in cardiac magnetic resonance imaging (MRI) for measurements of left ventricular dimensions in comparison with conventional gradient echo acquisition. Thirty-one subjects with a variety of left ventricular morphologies to represent a typical clinical population were studied. Short-axis data sets of the left ventricle (LV) were acquired using a conventional turbo-gradient echo and an ultrafast hybrid gradient echo/echo planar sequence with acquisition in real-time. End-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF) and left ventricular mass (LV mass) were measured. The agreement between the two acquisitions and interobserver, intraobserver and interstudy variabilities were determined. The bias between the two methods was 5.86 ml for EDV, 0.23 ml for ESV and 0.94% for EF. LV mass measurements were significantly lower with the real time method (mean bias 14.38 g). This is likely to be the result of lower spatial resolution and chemical shift artefacts with the real-time method. Interobserver, intraobserver and interstudy variabilities were low for all parameters. In conclusion, real time acquisition in MRI can provide accurate and reproducible measurements of LV dimensions in subjects with normal as well as abnormal LV morphologies, but LV mass measurements were lower than with conventional gradient echo imaging. PMID- 11502425 TI - Potential use of the undersampling technique in the acquisition of nuclear magnetic resonance signals. AB - This communication reviews the use of undersampling techniques to acquire NMR signals. Undersampling transforms bandpass free induction decay (FID) signals, centered at high frequencies, into lowpass signals or bandpass signals centered at much lower frequencies. Consequently, the analog electronic stages that perform the demodulation can be eliminated, gaining in stability and reducing the phase distortion while maintaining an equivalent or better signal to noise ratio when an adequate sampling rate is chosen. The technique has been tested on a BRUKER BIOSPEC BMT 47/40, and the results show that undersampling could be used to process NMR and MRI signals, extending the range of applications of the 'digital radio' techniques to NMR and MRI apparatus. PMID- 11502426 TI - Creatine loading and resting skeletal muscle phosphocreatine flux: a saturation transfer NMR study. AB - 31P saturation-transfer nuclear magnetic resonance spectroscopy was used to study skeletal muscle phosphocreatine (PCr) flux in healthy male volunteers. Data analysis included consideration of effects from incomplete saturation and radiofrequency spillover. Spectra were recorded from the resting gastrocnemius muscle before and after 6 days of creatine monohydrate (Cr-H2O) intake (20 g/day). Parallel to an improved muscle performance during maximal intermittent exercise following Cr-H2O supplementation, the concentration of PCr increased (P=0.01) by 23% (34.9+/-2.8 mmol/l vs. 28.6+/-2.7 mmol/l), whereas other metabolites were unaffected (inorganic phosphate: 4.3+/-1.4 mmol/l, free intracellular Mg(2+): 1.1+/-0.7 mmol/l, cytosolic pH: 7.04+/-0.02). Forward and reverse fluxes through the creatine kinase (CK) reaction did not change significantly from their baseline levels (v(for): 11.8+/-5.4 mmol/l per second vs. 15.3+/-6.8 mmol/l per second, (v(rev): 9.5+/-3.4 mmol/l per second vs. 10.9+/ 3.7 mmol/l per second). The rate of PCr resynthesis in resting muscle is not limited by the CK reaction, which is near equilibrium. Consequently, the post load increase in total creatine has no effect on the unidirectional CK reaction rates. PMID- 11502427 TI - Regional metabolite levels of the normal posterior fossa studied by proton chemical shift imaging. AB - MR spectroscopy of the posterior fossa is pitted with numerous technical difficulties. It is, however, of great clinical interest in the study of the degenerative diseases and tumors of this area. We have developed a method to perform 2D CSI of this area, by using a sagittal slice and a careful positioning of outer volume saturation. We performed this acquisition in 30 healthy volunteers to determine the normal metabolic ratios in five voxels of this area (mesencephalon, pons, medulla oblongata, vermis, cerebellar white matter). The main technical difficulty was magnetic field inhomogeneity in the lower brainstem generated by dental alloys. However, 88% of the voxels were of sufficient quality to be analyzed. The statistically significant regional variations were a higher NAA/Cr ratio in the pons than in the medulla oblongata, higher Cho/Cr in the pons than in the mesencephalon and higher Cho/Cr in the cerebellar white matter than in the vermis. We conclude that 2D CSI of the brainstem, although technically delicate can be performed in most patients. PMID- 11502428 TI - Concordance of information in parallel electronic and paper based patient records. AB - OBJECTIVE: to evaluate the results of parallel use of both paper based and electronic patient records with respect to concordance of corresponding information in two continuously updated versions of the same records. DESIGN: retrospective evaluation of patient records, comparing documentation in electronic and paper based patient records. SETTING: Department of Neurology in a Norwegian university hospital using paper based and electronic patient records in parallel during migration towards completely electronic patient records. MATERIAL: electronic and paper based patient records of 90 randomly selected patients visiting the department between 1 November 1997 and 30 April 1999. RESULTS: seven percent of the electronic documents were significantly different in some way from the corresponding paper documents. About 4-13% of the documents in the electronic record were missing; one percent were missing from the paper record. CONCLUSION: parallel use of electronic and paper based patient records has resulted in inconsistencies between the record systems in our setting. Documentation is missing in both the electronic and paperbased records. When implementing electronic record systems intended to operate in parallel with paperbased systems, focus should be on securing the validity of all versions of the record. PMID- 11502429 TI - Moving time window aggregates over patient histories. AB - Moving window concepts are used in temporal query languages for aggregate functions over the dimension 'time'. In the medical domain, aggregation of patient data over time windows builds a powerful mechanism within clinical database queries to satisfy a class of typical medical question formulations. Contrary to other fields, like the business domain for example, there is the additional need to synchronize time windows with the individual course of diseases rather than with the calendar system only. In this paper, we present several variants of shifting time windows over patient histories and suggest a set of essential options for a moving window clause. The proposed parameters for window creation as well as suitable default settings are discussed in the context of retrieving data from medical records. PMID- 11502430 TI - Cognitive psychological studies of representation and use of clinical practice guidelines. AB - Clinical practice guidelines provide a means to enhance physician performance. This investigation was undertaken in an attempt to understand the nature of impact of guideline use on physician performance. We investigated the impact of (a) algorithmic-based and (b) text-based practice guidelines on clinical decision making by physicians at varying levels of expertise. Data were collected using clinical scenarios and a think-aloud paradigm, both with (primed) and without (spontaneous) the use of the guidelines. The two guidelines used in the study were management of diabetes and screening for thyroid disease. The results show that guidelines were used as reminders for both experts and non-experts. Guidelines acted as an educational tool for non-experts by assisting in knowledge reorganization, particularly for the non-experts. Text and algorithmic guideline formats were both useful to physician performance depending on the purpose of use: solving clinical problems or learning. These results provide insights into how guidelines can be fine-tuned for different users and for different purposes. Empirical research, coupled with design principles from the cognitive sciences, can form an essential component of guideline design and development. PMID- 11502431 TI - Implementation of clinical guidelines through an electronic medical record: physician usage, satisfaction and assessment. AB - CONTEXT: We developed and evaluated the Emergency Department Expert Charting System (EDECS) to provide real-time guidance regarding the care of low back pain in adults, fever in children, and occupational exposure to blood and body fluids in health care workers, by embedding clinical guidelines within an electronic medical record. OBJECTIVE: To describe the behaviors and attitudes of physicians who used EDECS. DESIGN: Pre-post questionnaires were used to assess physician attitudes. Time studies of the intervention phase were observational, using clocks embedded in the software. PARTICIPANTS: One hundred and forty two residents and interns in emergency, pediatric, internal, and family medicine and patients with the above-mentioned complaints. MAIN OUTCOME MEASURES: Physician utilization of EDECS, time spent using EDECS, physician satisfaction and beliefs. RESULTS: Eighty four percent of the 142 eligible physicians used EDECS at least once. Five hundred and ninety one of 789 (75%) eligible cases were completed using EDECS. Median session time decreased from 12 min for session 1, to 5.5 min for sessions 16 and above. Physicians generally agreed that care with EDECS was better than standard care, particularly with respect to documentation. There was, however, considerable heterogeneity in belief among complaints. CONCLUSIONS: These data illuminate both the potentials of computer-assisted decision making and the need for context-specific approaches when attempting to implement guidelines. PMID- 11502432 TI - Indicators of organizational readiness for clinical information technology/systems innovation: a Delphi study. AB - The study presented in this article represents the second phase in a multi-phased research program focused on health care organization readiness for clinical information technology/system (IT/S) innovation. The overall purposes of this exploratory study were to: (1) validate the seven IT/S innovation readiness sub dimensions of a heuristic organizational information technology/systems innovation model (OITIM) developed in phase one of the research program, and (2) identify indicators to assess the validated sub-dimensions. The study was conducted with an expert panel using a two-round modified Delphi technique. In Round #1, panelists supported retention of the OITIMs' seven theoretical IT/S innovation readiness sub-dimensions with an interrater agreement level range of 82-100%. Their sub-dimension importance ratings ranged from 3.27 to 3.72 (1=not important to 4=critically important) with the resources sub-dimension receiving the highest rating of 3.72. Panelists recommended that two sub-dimensions, 'Staffing and Skills' and 'Operations', be renamed to 'End-Users' and 'Management Structures', respectively, and that one sub-dimension, 'Administrative Support' be added. In Round #2, panelists identified a total of 316 indicators to assess the eight sub-dimensions. A two-step thematic analysis process was done with these indicators to reduce duplication and overlap. In Step 1, the investigator created ten preliminary theme categories per sub-dimension. In Step 2, a coding team categorized 279 (88%) of the 316 indicators into preliminary sub-dimension themes to create an organizational IT/S innovation readiness assessment taxonomy. This preliminary taxonomy was used to develop an organizational information technology/systems innovation readiness scale that is currently being pilot tested in phase three of the research program. PMID- 11502433 TI - Information technology sophistication in health care: an instrument validation study among Canadian hospitals. AB - Several empirical studies have shown that the use of computer-based information systems could have positive impacts on organizational performance. Many agree to say that health care organizations are no exceptions. But if one wishes to identify the effects of information technology (IT) on the delivery of care, one must be able to characterize IT for operationalization purposes. The primary objective of this research project is to develop and validate a measurement instrument of IT sophistication in the hospital milieu. Such instrument should provide hospital managers with a diagnostic tool capable of indicating the profile of their respective institutions in regard to IT adoption and comparing this profile to those of other similar medical centers. Both qualitative and quantitative data were collected in order to assess the psychometric properties of the measurement instrument. Overall, findings suggest a high-moderate level of functional sophistication, a somewhat low level of technological sophistication, and an even lower level of integration sophistication in all of the sampled medical centers. Hence, future investments shall therefore be directed towards the integration of clinical and administrative applications and the acquisition of more advanced technological devices, more specifically those, which allow direct capture of clinical data at the bedside. PMID- 11502434 TI - Overview consensus statement. PMID- 11502435 TI - Is the flare phenomenon clinically significant? AB - OBJECTIVES: The existing luteinizing hormone-releasing hormone (LHRH) analogs have been the preferred method of inducing androgen deprivation for prostate cancer for over a decade. These agents are well known to cause a surge in serum testosterone levels during the first week of therapy. However, there are wide discrepancies in reports of the frequency and severity of acute clinical progression or clinical flare that might result from the testosterone surge. Also, there is not a clear consensus as to whether antiandrogens should be routinely given to all patients during the first month of LHRH therapy to prevent flare responses. METHODS: Clinical trials involving LHRH analog therapy for prostate cancer were reviewed, and the frequency of clinical flare responses noted. Particular attention was given to the kinds of clinical problems associated with the flare response. The use of LHRH analog therapy in treatment of patients with prostate cancer for indications other than overt metastatic disease is discussed, because this is becoming a much more common use of these agents. This article analyzes 2 placebo-controlled, double-blind trials testing the effectiveness of existing antiandrogens in ameliorating flare responses. RESULTS: The use of LHRH analogs for patients with stage D2 disease can be associated with clinical flare in approximately 10% of D2 patients. In addition to bone pain, cord compression, and bladder outlet obstruction, another potentially severe side effect is cardiovascular risk arising presumably from hypercoagulability associated with a rapid increase in tumor burden. In clinical series involving D2 patients, the frequency of clinical flare greatly varies, probably because of the level of scrutiny of the investigator and/or the prostate cancer tumor burden present at the initiation of therapy. Concomitant antiandrogen therapy reduces, but does not totally eliminate, the flare responses in patients at high risk for flare. Treating prostate cancer in the D0 stage or in the neoadjuvant setting will result in biochemical evidence of testosterone surge, but these patients are at very little risk for clinical flare responses. CONCLUSIONS: There is a wide variation in the reported frequency of clinical flare responses from LHRH analogs during the initial treatment of patients with stage D2 disease. The risk-to-benefit ratio, especially in patients with symptomatic bone metastasis, would dictate routine use of antiandrogen therapy for the first month of LHRH analog treatment. For patients at risk for cord compression, other means of ablating testosterone might be considered, such as ketoconazole, orchiectomy, or LHRH antagonists. Clinical flare responses, as opposed to biochemical flare responses, are very rare during LHRH analog therapy for stage D0 disease and/or in the setting of neoadjuvant hormonal therapy. PMID- 11502437 TI - Luteinizing hormone-releasing hormone monotherapy: a viable option for treatment of prostate cancer? AB - When addressing the appropriateness of luteinizing hormone-releasing hormone (LHRH) analog monotherapy as a method of androgen deprivation, it is important to look at the alternative option of androgen deprivation by combined androgen blockade (CAB). The randomized control trials studying CAB have been thoroughly analyzed and subjected to meta-analyses that have shown a small but significant difference in survival at 5 years. These findings have been subject to variable interpretations. Much like other treatment-option decisions for prostate cancer, it is mandatory to inform patients fully on the costs and benefits of monotherapy versus CAB and to incorporate the patient's concerns and preferences in the decision-making process. Currently, parameters to identify a subgroup of patients who might specifically benefit from a combined androgen deprivation treatment policy are not available. When using monotherapy, it is necessary to recognize several important facts. Castrate testosterone levels are not achieved or maintained in all patients. For some patients this may be a disadvantage. Additionally, before categorizing a patient as having an androgen-independent tumor, it is important to measure serum testosterone to ensure that LHRH analog monotherapy has achieved and maintained a castrate testosterone level. PMID- 11502439 TI - Antiandrogen monotherapy: a new form of treatment for patients with prostate cancer. AB - Nonsteroidal antiandrogens are generally used in conjunction with castration as combined androgen blockade. However, the changing profile of patients with prostate cancer has made monotherapy with a nonsteroidal antiandrogen an attractive alternative therapeutic approach, offering potential quality-of-life benefits over conventional treatment modalities. Of available antiandrogens, monotherapy with bicalutamide has been most extensively evaluated. Combined data from 2 studies at a median follow-up time of 6.3 years revealed no statistically significant difference in overall survival between bicalutamide 150-mg monotherapy and castration in patients with nonmetastatic locally advanced disease. In patients with metastatic disease, there was a statistically significant difference (6 weeks) in overall survival in favor of castration. Bicalutamide monotherapy is associated with significant quality-of-life benefits (sexual interest and physical capacity), with preliminary data suggesting that the risk of osteoporosis may also be reduced by bicalutamide 150-mg monotherapy compared with castration. In general, bicalutamide is well tolerated, with a predictable adverse-effect profile. Breast pain (40%) and gynecomastia (49%) are the most common adverse events seen during monotherapy with this drug. In summary, the availability of bicalutamide 150-mg monotherapy broadens treatment options for men with locally advanced prostate cancer, offering a viable and attractive alternative to castration in this patient population. Ongoing studies will determine the role of bicalutamide in the treatment of localized disease. PMID- 11502441 TI - Luteinizing hormone-releasing hormone antagonists in prostate cancer. AB - Luteinizing hormone-releasing hormone (LHRH) antagonists work by directly inhibiting LHRH without any initial stimulation of the LHRH receptor. The physiologic response is a direct and rapid decrease in luteinizing hormone, follicle-stimulating hormone, and testosterone without any flare. Although there has been extensive basic-science work on these medications, practical shortcomings have limited clinical studies in prostate cancer. Many of these compounds induce significant histamine-mediated side effects, and until recently, no depot form existed. In 2 recent phase-3 studies comparing abarelix depot with leuprolide and with leuprolide plus bicalutamide, abarelix lowered serum testosterone more quickly. None of the 89 patients on leuprolide alone were castrate on day 8 as opposed to 72% of the 180 patients randomized to abarelix (P <0.001). Similarly, none of the combination group were castrate by day 8, whereas 68% of the abarelix patients were castrate (P <0.001). In addition, 82% of the patients treated with leuprolide and 86% of those given leuprolide/bicalutamide had testosterone surge, whereas none of the abarelix patients did (P <0.001 for both studies). Both phase 2 and phase 3 data show abarelix to be well tolerated. In conclusion, LHRH antagonists offer the physiologic response of orchiectomy without surgery. These medications are well tolerated and a depot form now exists. The expansion of indications for androgen deprivation, such as downsizing or intermittent therapy, could provide many opportunities for their use. Despite these encouraging advances, however, their routine use for advanced prostate cancer may depend on demonstration of a survival advantage in avoiding flare. PMID- 11502443 TI - In vitro mechanism of PC SPES. AB - PC SPES (BotanicLab, Brea, California) an herbal supplement for patients with prostate cancer, is composed of 7 highly concentrated Chinese herbs and 1 US herb. It was developed in seeking positive attributes of Chinese and Western medicine for cancer treatment. Chemical standardization of this composition showed that baicalin is the most abundant active compound. Several reports on phase 2 clinical studies of PC SPES suggest that it is a well-tolerated active treatment for androgen-independent prostate cancer. In this report, data obtained from various laboratory experiments will be presented to elucidate the in vitro mechanism. Profound biologic effects of PC SPES on prostate cancer cells were observed on both androgen-dependent (LNCap) and androgen-independent (DU-145) cell lines. These effects include the following: (1) induction of cell apoptosis and cell cycle modulation; (2) inhibition of cell proliferation; (3) downregulation of bcl-2, bcl-6, proliferating cell nuclear antigen, and prostate specific antigen proteins; (4) downregulation of androgen receptor (AR); and (5) upregulation of p53, bax, and p21 proteins. Concurrent animal studies using 2 different models, Copenhagen rats and nude mice, confirmed a dose-dependent suppressive effect of PC SPES on tumor volumes and tumor progression. Our results show that the cytotoxic and cytostatic properties of PC SPES are not entirely dependent on the presence of AR. The antitumor mechanism of PC SPES is complex. It involves multiple metabolic pathways, such that the whole extract acts on redundant mechanisms, which otherwise will permit cell survival if a single target agent is used. PMID- 11502444 TI - The effects of phytotherapeutic agents on prostate cancer: an overview of recent clinical trials of PC SPES. AB - The clinical effects of the dietary supplement PC SPES will be reviewed. This herbal extract is a combination of 1 US herb and 7 Chinese herbs shown to be active against both androgen-dependent and androgen-independent prostate cancer. Anecdotal reports on the effectiveness of this agent have led to several clinical trials. These clinical trials have shown prostate-specific antigen partial response rates and overall improvements in quality of life. Although these studies have concluded that there is a possible estrogenic component, evidence exists that this is not the sole reason for the efficacy of PC SPES. At this point, clinical trials have validated PC SPES as an option for patients with prostate cancer, but further study is needed to pinpoint the exact mechanism of action. PMID- 11502448 TI - Early versus late androgen deprivation therapy in metastatic disease. AB - Patients presenting with metastatic prostatic cancer can be categorized into 3 groups. At present, most patients seen with metastases are those identified as having lymph-node disease when being assessed for curative therapy. The second group consists of patients with a high level of prostate-specific antigen, without symptoms, who are found incidentally to have asymptomatic bone metastases or metastases in soft tissue. The third group, who previously comprised about half of patients presenting with metastatic prostate cancer, are those presenting with painful metastases. There can be little doubt that most urologists will treat the second and third group of patients with hormone therapy at the outset. The question is whether the mere presence of lymph-node metastases or painless bony or soft tissue metastases justifies the side effects of long-term hormone therapy. A number of studies have shown a benefit in progression-free survival in the treatment of patients with lymph-node disease. Only 1 study has shown an advantage in overall survival. All studies of hormone therapy in asymptomatic and symptomatic metastatic disease have shown that serious complications of the disease can be avoided by offering hormonal therapy when the diagnosis is established. With the new generation of antiandrogens, differentiation therapies, and possibly alpha-reductase inhibitors, hormone therapy causes many fewer side effects than in the past and can be tolerated for longer periods of time. An aim of early hormonal therapy and its justification is a possible improvement in the quality of life of patients with metastatic prostate carcinoma, whose quantity of life cannot be lengthened. PMID- 11502446 TI - Use of antisense oligonucleotides targeting the antiapoptotic gene, clusterin/testosterone-repressed prostate message 2, to enhance androgen sensitivity and chemosensitivity in prostate cancer. AB - BACKGROUND: Androgen resistance develops, in part, from upregulation of antiapoptotic genes after androgen withdrawal. Identification and targeting of genes mediating androgen-independent (AI) progression may lead to development of novel therapies that delay hormone-refractory prostate cancer. Clusterin is a cell survival gene, that increases after androgen ablation. Here, we review clusterin's functional role in apoptosis and the ability of antisense oligonucleotides (ASOs) against clusterin to enhance apoptosis in prostate cancer xenograft models. RESULTS: Immunostaining of radical prostatectomy specimens confirm that clusterin is highly expressed in 80% prostate cancer cells after neoadjuvant hormone therapy, but is low or absent (<20%) in untreated specimens. Clusterin levels increase >10 fold in regressing Shionogi tumors after castration. Pretreatment of mice bearing androgen-dependent Shionogi tumors with calcium antagonists inhibited castration-induced apoptosis, tumor regression, and clusterin gene upregulation, illustrating that clusterin is an apoptosis associated gene and not an androgen-repressed gene. Clusterin ASOs reduced clusterin levels in a dose-dependent and sequence-specific manner. Adjuvant treatment with murine clusterin ASOs after castration of mice bearing Shionogi tumors decreased clusterin levels by 70% and resulted in earlier onset and more rapid apoptotic tumor regression, with significant delay in recurrence of AI tumors. Species-specific clusterin ASOs also increased the cytotoxic effects of paclitaxel, reducing the 50% inhibitory concentration (IC(50)) of PC-3 and Shionogi cells by 75% to 90%. Although clusterin ASOs had no effect on the growth of established AI Shionogi or PC-3 tumors, clusterin ASOs synergistically enhanced paclitaxel-induced tumor regression in both Shionogi and PC-3 models. CONCLUSIONS: Collectively, these data identify clusterin as an antiapoptosis protein, upregulated in an adaptive cell-survival manner by androgen ablation and chemotherapy, which confers resistance to various cell-death triggers. Inhibition of clusterin upregulation using clusterin ASOs can enhance cell death after treatment with androgen ablation and chemotherapy. PMID- 11502450 TI - Androgen deprivation therapy for patients with clinically localized (stages T1 to T3) prostate cancer and for patients with biochemical recurrence after radical prostatectomy. AB - Recently published studies suggest a benefit for androgen deprivation therapy (ADT) delivered early in the course of prostate cancer. However, the use of ADT specifically in patients with clinically localized disease or biochemical-disease recurrence after local therapy is not well defined. Potential candidates for primary ADT include patients who are poor candidates for definitive local therapy because of advanced age or comorbid conditions, as well as patients with significant local disease who refuse standard therapy. Treatment strategies designed to minimize the side effects of prolonged therapy, such as intermittent ADT or antiandrogen monotherapy, show promise as alternatives to continuous ADT in some patients. The role of ADT in patients with clinically localized and recurrent prostate cancer, whether it is delivered in a continuous or intermittent fashion, must be determined in randomized, prospective trials. PMID- 11502452 TI - The role of androgen ablation in patients with biochemical or local failure after definitive radiation therapy: a survey of practice patterns of urologists and radiation oncologists in the United States. AB - To identify therapeutic patterns for putative prostate cancer treatment failures and the role played by androgen ablation therapy in these patients, a questionnaire study was undertaken with urologists and radiation oncologists who had attended a brachytherapy forum at the Seattle Prostate Institute (SPI). Hypothetical questions were asked about recommendations the physicians would give to a patient demonstrating biochemical or local failure after external-beam radiation therapy. Most of the physicians queried were in private practice; 53% were radiation oncologists and 47% were urologists. The respondents' recommendations for a hypothetical patient, who was 45 to 65 years of age, with a biopsy-proven local recurrence was treatment with androgen ablation (35% of respondents), radical prostatectomy (25%), interstitial brachytherapy (20%), and observation (19%). In the 65- to 75-year-old patient with a local recurrence, the respondents recommended observation (43%), androgen ablation (35%), interstitial brachytherapy (17%), and radical prostatectomy (4%). In patients receiving androgen ablation for a biochemical failure alone, there was no consensus on whether to use luteinizing hormone-releasing hormone agonist alone, total androgen ablation, orchiectomy, or intermittent androgen ablation. Criteria that prompted physicians to initiate androgen ablation were based on the rate of prostate-specific antigen (PSA) increase (67%), an absolute PSA number (24%), or clinical failure (9%). In the younger patient with a local recurrence, local intervention with radical prostatectomy or interstitial brachytherapy was recommended most often, followed by androgen ablation, then by observation. In the older patient, observation was recommended most often, followed closely by androgen ablation. Overall, there was a lack of consensus on how to deliver androgen ablation. However, there was remarkable agreement between urologists and radiation oncologists on virtually all issues queried. PMID- 11502453 TI - Duration of neoadjuvant androgen deprivation therapy before radical prostatectomy and disease-free survival in men with prostate cancer. AB - There is little evidence that neoadjuvant androgen deprivation therapy (ADT) of 3 months' duration before radical prostatectomy (RP) favorably influences disease free survival. However, recent data suggest that prolonged treatment may improve outcome. We conducted a prospective cohort study to determine whether ADT of either standard or prolonged duration before RP influences the risk of prostate specific antigen (PSA) failure. We followed 756 men treated for prostate cancer by RP between 1991 and 1998 in Quebec City. Of these, 240 received combined neoadjuvant ADT for either /=93 days (111 men), and 516 were treated by RP alone. Multivariate Cox regression was used to estimate the hazard ratios (HR) of PSA failure (>0.3 ng/mL) associated with treatment regimen controlling for age, clinical stage, grade, and initial PSA level. The median duration of follow-up was 4 years. Compared with men treated by RP alone, those who received neoadjuvant ADT for >/=93 days had an HR of PSA failure of 0.60. The inverse association with the risk of PSA failure became statistically significant from the third year on, reached its greatest magnitude after 4 years, and was still present 8 years after RP. No association was observed for ADT of /=1 high-risk features. PMID- 11502457 TI - Androgen deprivation therapy for prostate cancer chemoprevention: current status and future directions for agent development. AB - Prostate cancer chemoprevention is defined as the administration of natural and synthetic agents that inhibit >/=1 steps in the natural history of prostate carcinogenesis. The goal is to find agents that modulate the progression from normal epithelium to dysplasia to high-grade prostatic intraepithelial neoplasia (HGPIN) to locally invasive cancer and systemic disease. Another important goal for chemoprevention is the maintenance of an androgen-sensitive clinical state and delay of the emergence of androgen independence. There is a strong rationale for androgen deprivation therapy (ADT) as a chemoprevention strategy for prostate cancer based on evidence from epidemiologic, experimental, molecular pathophysiologic, and randomized, controlled clinical trials. This includes the fact that HGPIN, the most likely precursor of invasive cancer, is androgen dependent and responds to ADT. Although the large, phase-3 Prostate Cancer Prevention Trial (PCPT) of finasteride versus placebo has established the feasibility and role of ADT for primary prevention, nevertheless, limitations of the anticipated treatment-effect size (eg, 25% reduction) and the potential for selection of androgen resistance provide incentive for finding other effective chemopreventive agents. The availability of novel noncytotoxic pharmaceutical and natural products in clinical development create opportunities for improving the therapeutic index through the principles of combination therapy. The emergence of new powerful tools, such as gene chip complementary DNA microarrays for multiplex gene expression profiling, will accelerate the identification of new molecular targets and the design of rational combinations. Several agent classes have a strong basis for combination with ADT, including antiproliferatives, antioxidant micronutrients (selenium), antiestrogens, and nonsteroidal anti-inflammatory drugs (selective cyclooxygenase-2 inhibitors). PMID- 11502458 TI - Effect of androgen deprivation therapy on prostatic intraepithelial neoplasia. AB - There is a marked decrease in the prevalence and extent of high-grade prostatic intraepithelial neoplasia (PIN) in men with prostate cancer after androgen deprivation therapy (ADT) when compared with untreated cases. Basal cell hyperplasia, cytoplasmic clearing, and prominent atrophy of benign acini, with decreased ratio of acini to stroma, accompany this decrease. These findings indicate that the benign and dysplastic prostatic epithelium is androgen dependent. In the normal prostatic epithelium, luminal secretory cells are more sensitive to the absence of androgen than basal cells, and the proliferative cells of high-grade PIN share this androgen sensitivity. The loss of some normal, hyperplastic, and dysplastic epithelial cells with ADT is probably because of acceleration of programmed single-cell death. Remarkably little is known about the comparative effect of different forms of chemical ADT on PIN and cancer, although there appears to be a limited and consistent repertoire of morphologic responses to all forms of this therapy. Conversely, blockade of 5alpha-reductase with finasteride has little or no effect on PIN (or benign epithelium and cancer), unlike other forms of ADT. A recent international consensus conference sponsored by the World Health Organization concluded that identification of high grade PIN offered the possibility of chemoprevention with hormonal therapy to block the development of clinical cancer. Multiple chemoprevention trials are planned or under way to address this hypothesis. PMID- 11502459 TI - The effect of androgen deprivation therapy on health-related quality of life in men with prostate cancer. AB - Treatment for prostate cancer has a significant impact on health-related quality of life (HRQOL). We examined HRQOL in a cohort of men receiving androgen deprivation therapy (ADT) and a cohort who opted for surveillance. The cohort consisted of 1178 newly diagnosed patients from the Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE) database (a national longitudinal registry of patients with prostate cancer). General and disease-specific HRQOL outcomes were measured with validated instruments (the SF-36 and University of California at Los Angeles [UCLA] Prostate Cancer Index) at study entry and quarterly thereafter. Individuals were grouped by initial treatment: ADT, surveillance, radical prostatectomy, or radiation therapy. There were 106 men who selected surveillance, 167 men receiving ADT, 351 men treated by radical prostatectomy, and 75 men receiving radiation therapy in the first year after diagnosis. Mean age at diagnosis was 73 years of age, with surveillance patients the oldest and radical prostatectomy patients the youngest. Men receiving ADT reported poorer urinary and sexual function and a higher rate of urinary and sexual bother than patients selecting surveillance. ADT and surveillance HRQOL scores remained low (ie, poorer function) in the year after treatment, whereas men undergoing radical prostatectomy showed improvement in these scales. Patients who received ADT had reduced energy, poorer sexual and urinary function, and were more bothered by their urine and sexual function than patients undergoing other treatments, except surveillance. Longer follow-up time after start of ADT and surveillance is needed to discern the impact of comorbidities on HRQOL. PMID- 11502461 TI - Osteoporosis due to androgen deprivation therapy in men with prostate cancer. AB - OBJECTIVES: The frequency of osteoporotic fractures is greatly increased in men receiving androgen deprivation therapy (ADT), but whether the risk of osteoporosis differs between different types of ADT or between continuous and intermittent therapy has not been determined. Techniques for modifying ADT associated bone loss have not been clearly identified. METHODS: Risk factors for the development of osteoporosis in men receiving ADT will be reviewed. Relations between bone mineral density (BMD) values and the development of osteoporotic fractures, along with methods for preventing both BMD loss and osteoporotic fractures, will be discussed. RESULTS: ADT rapidly accelerates bone loss among men with prostate cancer and multiplies the risk of osteoporotic fractures among them. Factors other than ADT-associated bone loss contributing to this fracture risk include both decreased BMD before ADT and an increased tendency to fall associated with muscle weakness, impaired balance, and postural hypotension. Each of these factors may be associated with poor nutrition, advancing malignant disease, hypogonadism of non-ADT origin, advanced age, and the use of narcotic, antihypertensive, or sedative medications. Although the success of therapy designed to improve BMD values and lower the fracture rate in these patients has not been explored, regular exercise, smoking abstinence, adequate calcium, protein, and vitamin D intake, maintenance of weight, and the use of bisphosphonates or calcitonin may each have a useful therapeutic role. Theoretical considerations suggest that intermittent ADT may decrease the frequency of ADT-associated osteoporosis. CONCLUSIONS: An urgent need exists for the definition of techniques useful in preventing osteoporotic fractures in men receiving ADT for prostate cancer. PMID- 11502463 TI - The role of diethylstilbestrol in the treatment of prostate cancer. AB - OBJECTIVES: Diethylstilbestrol administration was a classic form of androgen deprivation therapy (ADT) that gradually fell out of favor because of its cardiovascular toxicity, economic disinterest on the part of manufacturers, and the emergence of novel therapeutic agents with a superior safety profile. The cost of contemporary agents and the efficacy of diethylstilbestrol (DES) have perpetuated the evaluation of this agent, especially in the circumstance of early hormone-refractory (clinical stage D2.5) disease. The objective of this study is to evaluate the status of DES-based therapies, and assess their efficacy and toxicity as a viable form of ADT. METHODS: Current research from single institution and group studies, as well as basic scientific investigations related to DES, were assessed with regard to the population studied, dosage, criteria for response, response rate, duration of response, and toxicity. RESULTS: Contemporary basic research has demonstrated a direct apoptotic effect of DES on prostate cancer cells. There is also evidence to support the ability of DES to suppress testosterone production at extratesticular sites and inhibit dihydroepiandrosterone sulfate serum levels. Contemporary cooperative group trials for stage D2 disease incorporating a DES arm have demonstrated therapeutic efficacy and equivalence to orchiectomy, which is marred by significant cardiovascular toxicity. In smaller single-institution studies (n = 17 to 38) of patients with D2.5 disease, an average response rate of 55% is noted with a mean time to clinical progression of 6.4 months (2 to 18). Cardiovascular toxicity occurred in 10% to 30% of patients, with events including deep vein thrombosis, myocardial infarction, and transient ischemic attack. Edema and gynecomastia was also noted. Strategies to reduce thromboembolic events, such as dose reduction or the use of warfarin sodium were unsuccessful, whereas the use of low-dose aspirin (100 mg daily) resulted in only 1 of 38 vascular events. CONCLUSIONS: In contemporary studies of DES as an agent for ADT in D2.5 patients, a reasonable response rate (40% to 60%) of modest duration (5 to 8 months) is noted. Cardiovascular complications still persist, requiring the development of safe, effective antithrombolic therapy to take advantage of this phenomenon. PMID- 11502465 TI - Growth factors and their receptors: new targets for prostate cancer therapy. AB - Stimulation of the signal transduction pathway of the epidermal growth-factor receptor (EGFR) tyrosine kinase family of receptors in tumor cells enhances cellular proliferation, prevents apoptosis, and promotes tumor-cell mobility, adhesion, and invasion. Therapeutic approaches used to target the EGFR and its signal transduction cascade include (1) monoclonal antibodies (eg, cetuximab [IMC C225]) directed against the extracellular binding domain of the receptor; and (2) trastuzumab, a monoclonal antibody binding to the HER2 receptor; immunotoxin conjugates use an antibody directed against EGFR joined to a cell toxin. All are in clinical trials for a number of cancers, including prostate cancer. Antisense strategies are in preclinical development. Low-molecular-weight inhibitors of the EGFR tyrosine kinase also in clinical development include OSI-774, PD182905, PKI 166, CI-1033, and ZD1839. ZD1839 has shown encouraging results in patients with prostate cancer in phase 1 trials. mn PMID- 11502466 TI - Rationale for combination ketoconazole/ vitamin D treatment of prostate cancer. AB - The high rate of progression of prostate cancer after androgen deprivation therapy mandates that new strategies be developed. Adjuvant therapy combined with androgen deprivation may slow or prevent progression. Ketoconazole plus calcitriol therapy is an example of 1 such a combination with a mechanistic basis for synergistic activity. Ketoconazole is commonly used as a second-line androgen deprivation therapy. This imidazole derivative is an inhibitor of P-450 enzymes, including those involved in steroidogenesis. Other P-450 enzymes that are inhibited by ketoconazole include 1alpha-hydroxylase and 24-hydroxylase, which metabolize vitamin D. Growth inhibition of prostate cancer cells by vitamin D depends on levels of the active metabolite, 1,25-dihydroxyvitamin D(3) (calcitriol). The enzyme 24-hydroxylase converts calcitriol to less active products. The inhibition of 24-hydroxylase by ketoconazole maintains the magnitude and duration of response to calcitriol. Combined ketoconazole/calcitriol therapy might therefore potentiate the antitumor activity of calcitriol. Because androgen-independent prostate cancer cells often remain responsive to growth inhibition by calcitriol, it is also possible that calcitriol would slow or prevent development of androgen-independent cancer growth. Another consideration is that ketoconazole blocks 1alpha-hydroxylase activity, which is the key enzyme that creates calcitriol in the body. Therefore, patients receiving ketoconazole therapy are likely to be deficient in vitamin D. The detrimental consequences of vitamin D deficiency in these patients would also be alleviated by the addition of calcitriol to the therapeutic regimen. PMID- 11502467 TI - The role of cyclooxygenase-2 in prostate cancer. AB - Cyclooxygenase-2 (COX-2) is the inducible isozyme of COX, a key enzyme in the conversion of arachidonic acid to prostaglandins and other eicosanoids. COX-2 is highly expressed in a number of human cancers and cancer cell lines, including prostate cancer. We studied the immunohistochemical expression of COX-2 in the human prostate gland. The enzyme is strongly expressed in smooth muscle cells of both the normal and cancerous prostate. Its expression in noncancerous epithelial cells is limited to the basal cell layer. In prostatic inflammation, luminal epithelial cells surrounded by lymphocytes are induced to express the enzyme. COX 2 is expressed in the epithelial cells of high-grade prostatic intraepithelial neoplasia and cancer. We have demonstrated that treatment of human prostate cancer cell lines with a selective COX-2 inhibitor induces apoptosis both in vitro and in vivo. The in vivo results also indicate that the COX-2 inhibitor decreases tumor microvessel density and angiogenesis. COX-2 inhibitors can prevent the hypoxic upregulation of a potent angiogenic factor, vascular endothelial growth factor. These results indicate that COX-2 inhibitors may, therefore, serve as effective chemopreventive and therapeutic agents in cancer of the prostate. PMID- 11502468 TI - In vivo suicide gene therapy model using a newly discovered prostate-specific membrane antigen promoter/enhancer: a potential alternative approach to androgen deprivation therapy. AB - Prostate-specific membrane antigen (PSMA) is a type-2 membrane protein expressed in the prostate, and it is highly expressed in metastatic or poorly differentiated adenocarcinomas. Moreover, PSMA expression is upregulated by androgen deprivation. These advantages make PSMA a useful target for prostate cancer therapy, especially in combination with conventional hormonal treatment. We recently reported that a prostate-specific enhancer is present in the third intron of the PSMA gene. In this study, we have further analyzed the activity of PSMA promoter/enhancer in prostate cancer cells and cells of other tissue origins (breast cancer MCF-7, lung cancer H157, and colorectal cancer HCT8 cells), and we have examined whether this construct could be used for efficient expression of the suicide gene, cytosine deaminase (CD), in vivo. The PSMA promoter/enhancer expressed the luciferase reporter gene in the prostate cancer lines LNCaP and C4 2, with 8- to 20-fold higher expression than the simian virus 40 promoter/enhancer, although it was inactive in the other cell lines. This construct efficiently drove the suicide gene CD, sensitizing C4-2 cells to 5 fluorocytosine (5-FC) with the inhibitory concentration (IC(50)) <300 micromol/L in vitro. Athymic male nude mice bearing the transfected C4-2 cells were treated with intraperitoneal injections of either 5-FC (600 mg/kg) twice a day or saline solution for 3 weeks. C4-2 cell tumors were eliminated by 5-FC when they were expressing our therapeutic construct carrying CD under the regulatory control of the PSMA promoter/enhancer. Our results show the in vivo utility of the PSMA promoter/enhancer in a gene therapy situation targeting prostate cancer. PMID- 11502469 TI - Understanding bone strength: size isn't everything. PMID- 11502470 TI - Modulation of appositional and longitudinal bone growth in the rat ulna by applied static and dynamic force. AB - Appositional and longitudinal growth of long bones are influenced by mechanical stimuli. Using the noninvasive rat ulna loading model, we tested the hypothesis that brief-duration (10 min/day) static loads have an inhibitory effect on appositional bone formation in the middiaphysis of growing rat ulnae. Several reports have shown that ulnar loading, when applied to growing rats, results in suppressed longitudinal growth. We tested a second hypothesis that load-induced longitudinal growth suppression in the growing rat ulna is proportional to time averaged load, and that growth plate dimensions and chondrocyte populations are reduced in the loaded limbs. Growing male rats were divided into one of three groups receiving daily 10 min bouts of static loading at 17 N, static loading at 8.5 N, or dynamic loading at 17 N. Periosteal bone formation rates, measured 3 mm distal to the ulnar midshaft, were suppressed significantly (by 28-41%) by the brief static loading sessions despite normal (dynamic) limb use between the daily loading bouts. Static loading neither suppressed nor enhanced endocortical bone formation. Dynamic loading increased osteogenesis significantly on both surfaces. At the end of the 2 week loading experiment, loaded ulnae were approximately 4% shorter than the contralateral controls in the 17 N static and dynamic groups, and approximately 2% shorter than the control side in the 8.5 N static group, suggesting that growth suppression was proportional to peak load magnitude, regardless of whether the load was static or dynamic. The suppressed growth in loaded limbs was associated with thicker distal growth plates, particularly in the hypertrophic zone, and a concurrent retention of hypertrophic cell lacunae. Negligible effects were observed in the proximal growth plate. The results demonstrate that, in growing animals, even short periods of static loading can significantly suppress appositional growth; that dynamic loads trigger the adaptive response in bone; and that longitudinal growth suppression resulting from compressive end-loads is proportional to load magnitude and not average load. PMID- 11502471 TI - Role of serum leptin, insulin, and estrogen levels as potential mediators of the relationship between fat mass and bone mineral density in men versus women. AB - Although fat mass is related to bone mineral density (BMD), the potential mechanism(s) of this effect remain to be defined. Thus, we assessed the role of the candidate hormones, leptin, insulin, and estrogen in mediating fat mass effects on the skeleton. Specifically, we related these hormones and fat mass to BMD at the total hip, mid-lateral spine, and mid-distal radius in a sample of 137 premenopausal women (age range 21-54 years), 165 postmenopausal women (34-93 years), and 343 men (23-90 years) recruited from the general population. Fat mass and BMD were significantly related in pre- and postmenopausal women at multiple sites, whereas this relationship was only weakly present in men at the total hip. Serum leptin levels were also significantly related to BMD in the women, but not in the men. Insulin was associated with hip BMD in the women, and bioavailable estradiol (E2) was correlated with BMD at all sites in men and in postmenopausal women. In the women, adjusting for leptin reduced the strength of the association between fat mass and BMD, with further adjustments for insulin or bioavailable E2 having no additional effects. Adjusting for leptin in the men had no consistent effect on the relationship between fat mass and BMD. Collectively, these data suggest that there is a sexual dimorphism in the relationship of fat mass and leptin to BMD, with both being positively associated with BMD in women but not in men. In women, leptin may also mediate at least part of the protective effect of fat mass on the skeleton. PMID- 11502472 TI - Bone biomechanical properties in prostaglandin EP1 and EP2 knockout mice. AB - Prostaglandins play an important role in regulating the bone adaptation response to mechanical stimuli. Prostaglandin E2 (PGE2) is an effective modulator of bone metabolism. Administration of PGE2 to rodents results in increased cancellous and cortical bone mass translating into enhanced mechanical strength. The PGE2 influence on bone is mediated through four well-characterized receptors (EP1, EP2, EP3, and EP4). Although the PGE2 pathways and mechanisms of action on cells involved in bone adaptation are still under investigation, it is now known that each receptor plays a unique role in regulating PGE2-related bone cell function. The EP1 subtype is coupled with Ca2+ mobilization. The EP2 subtype stimulates cyclic adenosine monophosphate (cAMP) formation. cAMP in turn is responsible for the early cellular signal that stimulates bone formation. This study compared physical and biomechanical properties of bone in EP1 and EP2 knockout mice to their corresponding wild-type controls. Ash weight was measured in the ulnae, and femurs and vertebral bodies were tested in three-point bending and compression, respectively. The results suggest: (a) EP1 receptors have a minimal influence on skeletal strength or size in mice; and (b) EP2 receptors have a major influence on the biomechanical properties of bone in mice. The absence of EP2 receptors resulted in weak bone biomechanical strength properties in the EP2 knockout model as compared with the corresponding wild-type control mice. PMID- 11502473 TI - Bipedal stance exercise enhances antiresorption effects of estrogen and counteracts its inhibitory effect on bone formation in sham and ovariectomized rats. AB - In this study we employed a raised cage model in combination with estrogen to observe their effects on the proximal tibial metaphysis (PTM) and tibial shaft (TX) in sham-operated or ovariectomized rats. A total of 105 6-month-old female Sprague-Dawley rats were used in the study. Bilateral sham ovariectomy or ovariectomy was performed at day 0 and the rats were housed in normal height or raised cages (RCs) and injected subcutaneously twice per week with 10 microg/kg of 17beta-estradiol (E2) or vehicle for 4 and 8 weeks. Because the time course of bone loss or bone gain distribution was not uniform in the metaphyses of the tibia, we subdivided the PTM into three zones (medial, central, and lateral) to observe the different bone loss or bone gain patterns after ovariectomy and/or raised cages. We found that: (1) E2 alone did not alter bone area or architecture in sham rats, whereas RC alone increased trabecular thickness and area of PTM, but had no effects on TX; (2) Ovx induced most bone loss from the central zone of the PTM and endocortical surface of TX, accompanied by decreased trabecular number and increased bone resorption; (3) E2 alone prevented ovx-induced bone loss by preserving trabecular number and depressing bone resorption; (4) RC alone partially compensated for bone loss following ovx by thickening the surviving trabeculae in lateral and medial zones, and tended to stimulate bone formation and decrease bone resorption; and (5) RC plus E2 increased trabecular bone area by having an additive effect on bone resorption and bone turnover. RCs helped to prevent the depressive effect of estrogen on periosteal bone formation. In conclusion, early and rapid bone loss occurred in the central zone of the metaphysis and endocortical surface after ovx. Estrogen replacement therapy prevented this loss. Raised cages partially compensated for bone loss following ovx by thickening the trabeculae in the lateral area of the metaphysis and decreased endocortical erosion. Combination treatment added bone to the PTM and prevented the decrease of periosteal bone formation after estrogen administration. PMID- 11502474 TI - Genetic variation in bone-regenerative capacity among inbred strains of mice. AB - Genetic variation in bone-regenerative capacity has not been studied in any animal model system. We developed a "drill-hole" model in the tail vertebra of inbred strains of mice that allows us to reproducibly introduce an injury with a defined boundary and quantify the rate of bone healing using the combination of high-resolution Faxitron X-ray imaging and the ChemiImager 4000 Low Light Imaging System. Using this model, we demonstrate that bone-regenerative capacity is a genetically controlled trait with an estimated heritability of 72%, and that it differs significantly among inbred strains of mice. Of the 12 inbred strains tested, Sencar/PtJ was identified as the most suitable model for the study of hard-tissue regeneration. This strain regained 73% of bone loss 30 days after injury, in contrast to the slow healer, CBA/J, which recovered only 25% of the bone loss during the same period. Bone-regenerative capacity was not correlated with soft-tissue-regenerative capacity, suggesting that different sets of genes may regulate soft- and hard-tissue regeneration. It was, however, significantly correlated with total bone mineral density (R = 0.49, p < 0.01), indicating that high bone density is associated not only with prevention of bone fracture, but also with promotion of bone regeneration. PMID- 11502476 TI - The E11 osteoblastic lineage marker is differentially expressed during fracture healing. AB - Given the complexity of the fracture healing process and the involvement of a multitude of cells, we hypothesize that a very large number of genes would be transcriptionally regulated during the repair process. To identify genes that play a role during bone regeneration in cellular events, such as proliferation, migration, adhesion, and differentiation, we employed differential display and compared mRNA populations isolated from postfracture (PF) day 3 calluses to those of intact (contralateral) rat femurs. One such gene whose expression was upregulated at PF day 3 is identified as the osteoblastic lineage marker, E11 antigen. E11 is a cell membrane protein localized predominantly on osteoblasts and osteocytes. In this study we show that E11 mRNA expression is consistently upregulated during fracture repair, with elevated levels (tenfold) seen as early as PF day 3. These high levels of expression were maintained throughout all of the early stages of repair examined (PF day 3-21 calluses) and paralleled those of another osteoblastic marker, osteopontin. Similarly, high protein levels were detected throughout the reparative phase of the callus, particularly in osteoblasts, preosteocytes, and osteocytes, localized within the newly made osteoid. No labeling was detected in fibroblasts, proliferating chondrocytes, or hypertrophic chondrocytes, consistent with previous results. Taken together, these results suggest that the E11 antigen is indeed a suitable marker for both osteoblasts and osteocytes and that it plays a significant role in osteogenesis. PMID- 11502475 TI - Male rodent model of age-related bone loss in men. AB - Osteoporosis is a common occurrence in aging men. There is currently no appropriate animal model for studying age-related bone loss in men. To determine whether male Sprague-Dawley (SD) rats experience bone loss with aging and whether this rodent model is appropriate for studying age-related bone loss in men, SD rats aged 1-27 months were examined at the L-4 vertebra, the left femoral neck, and the left proximal tibia using peripheral quantitative computed tomography (pQCT) densitometry. In the L-4 vertebra of the male SD rats, cortical bone mineral content (BMC), cortical bone mineral density (BMD), and cortical bone thickness (Ct.Th) increased to a maximum at about 4 months of age and then plateaued. Vertebral cortical BMC began to decrease after about 13 months and vertebral Ct.Th began to decrease after about 9 months. By 27 months of age, vertebral cortical BMC decreased by 26.1% (p < 0.0001) and vertebral Ct.Th decreased by 31% (p < 0.0001). Vertebral cancellous BMC and vertebral cancellous BMD increased to a maximum at about 3 months of age and then declined progressively with aging after a short plateau. From 3 to 27 months of age, vertebral cancellous BMC and vertebral cancellous BMD had decreased linearly by 35.4% (p < 0.0001) and 49.4% (p < 0.0001), respectively. Both vertebral periosteal and vertebral endocortical perimeters of the L-4 vertebra of the rats increased with aging. From 9 to 27 months of age, the percent increase of vertebral endocortical perimeter (19.8%, p < 0.0001) was higher than that of vertebral periosteal perimeter (7.4%, p < 0.0001). This process was associated with a decrease with aging in vertebral Ct.Th. In addition, cancellous bone in the femoral neck and the proximal tibia began to be lost at 9 months of age and, by 27 months of age, cancellous BMC and cancellous BMD decreased by 59.7% (p < 0.0001) and 58.4% (p < 0.0001), respectively, in the femoral neck and by 72.2% (p < 0.0001) and 71.4% (p < 0.0001), respectively, in the proximal tibia. To gain further insight into the effects of aging on cancellous bone in the L-4 vertebra, histomorphometry was done on the L-4 vertebral body of animals aged 3, 6, 9, 18, and 24 months after pQCT densitometry. From 3 months of age and thereafter, cancellous bone volume (BV/TV) decreased progressively and, by 24 months, there was a decrease of 35.7% (p < 0.0001). In the L-4 vertebra, single- and double labeled surfaces, mineral apposition rate (MAR), and bone formation rate (BFR/BS) decreased with aging. In conclusion, age-related bone loss in male SD rats started mostly from 9 months of age when bone growth had been completed. Aging male SD rats experience bone loss comparable to that seen in men. Thus, male SD rats represent an appropriate animal model of age-related bone loss in men. We recommend using male SD rats that are 9 months old as the starting age for age related bone loss. We also suggest using the L-4 vertebra and femoral neck as the clinically relevant bone sites for determining the cause of the loss of bone, and how and whether therapeutic agents could modulate age-related bone loss in men. PMID- 11502477 TI - Novel mode of processing and secretion of connective tissue growth factor/ecogenin (CTGF/Hcs24) in chondrocytic HCS-2/8 cells. AB - The synthesis, processing, and secretion of human connective tissue growth factor (CTGF/Hcs24) in a human chondrocytic cell line, HCS-2/8, were analyzed immunochemically. By metabolic pulse-labeling, chasing, and subsequent immunoprecipitation analyses, active synthesis of CTGF was observed not only in growing HCS-2/8 cells, but also in confluent cells. However, secretion and processing of CTGF were found to be regulated differentially, depending upon the growth status. During phases of growth, HCS-2/8 cells released CTGF molecules immediately without sequestering them within the cell layer. In contrast, after the cells reached confluence, the secretion slowed, resulting in an accumulation of CTGF in the cells or extracellular matrices (ECMs). Also, in confluent cell layers, a 10 kDa protein that was reactive to an anti-CTGF serum was observed. This CTGF-related small protein was not detected immediately after labeling, but gradually appeared within 6 h after chase, which suggests its entity as a processed subfragment of CTGF. Surprisingly, the 10 kDa protein was stable even 48 h after synthesis, and was not released by ECM digestion, suggesting an intracellular maintenance and function. Taken together, the behavior of CTGF in HCS-2/8 cells is remarkably different from that reported in fibroblasts, which may represent unique roles for CTGF in the growth and differentiation of chondrocytes. PMID- 11502478 TI - Effects of calcitonin, amylin, and calcitonin gene-related peptide on osteoclast development. AB - Amylin and calcitonin gene-related peptide (CGRP) are homologous 37 amino acid peptides that are found in the circulation. Both peptides belong to the calcitonin family. Similar to calcitonin, amylin and CGRP inhibit osteoclast activity, although they are much less potent than calcitonin. Calcitonin is known to act on the latter stages of osteoclast development, inhibiting the fusion of committed preosteoclasts to form mature multinucleated cells; however, whether or not calcitonin acts earlier in the formation of the precursor osteoclasts is controversial. The question of osteoclast development has never been examined with respect to amylin and CGRP. These issues are addressed in the present study. We studied the effects of calcitonin (salmon and rat), amylin (human and rat), and CGRP (human and rat) in mouse bone marrow cultures stimulated to generate osteoclasts using 1alpha,25-dihydroxyvitamin D3. Calcitonin dose-dependently decreased the numbers of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells as well as TRAP-positive mono-/binucleated cells at concentrations >10(-13) mol/L. Amylin and CGRP showed similar effects at concentrations >10(-9) mol/L. In addition, calcitonin substantially reduced the ratio of TRAP-positive multinucleated to mono-binucleated cells, indicating an effect on fusion of osteoclast precursors. The present data establish that this family of peptides not only acts on mature osteoclasts but also inhibits their development in bone marrow cultures. This activity is shared by amylin and CGRP. The much greater potency of calcitonin than amylin and CGRP is consistent with the action of these peptides being mediated by calcitonin receptors. PMID- 11502479 TI - The effects of transplantation of osteoblastic cells with bone morphogenetic protein (BMP)/carrier complex on bone repair. AB - We investigated the effects of transplantation of osteoblastic cells with a bone morphogenetic protein (BMP)/carrier complex on bone repair by in vitro and in vivo experiments. Poly-D,L-lactic-co-glycolic acid/gelatin sponge (PGS) was used as a carrier for cell transplantation. In the in vitro experiments, three cell types, C3H10T1/2 cells, MC3T3-E1 cells, and primary osteoblastic cells, isolated from newborn rat calvariae (ROB cells), were cultured for 2 weeks on PGS alone or PGS containing BMP-2 (PGS/BMP). C3H10T1/2 cells cultured on PGS/BMP expressed several markers related to differentiation of both osteoblasts and chondrocytes, such as alkaline phosphatase (ALP) activity and mRNAs for osteocalcin and aggrecan, whereas the cells cultured on PGS alone expressed no such markers. MC3T3-E1 cells cultured on PGS/BMP exhibited a more ALP-positive cells than those cultured on PGS alone. PGS/BMP promoted ROB cell differentiation into both osteoblasts and chondrocytes. In the in vivo experiments, we transplanted ROB cells, which had been cultured on PGS alone or PGS/BMP in vitro for 2 weeks, into bone defects created in rat calvariae. Transplantation of ROB cells cultured on PGS alone generated little new bone. Transplantation of ROB cells cultured on PGS, which absorbed a low dose (10 ng) of rhBMP-2,; induced significantly higher bone mineral content than PGS/BMP alone, although application of a high dose (1 microg) of rhBMP-2 induced no difference in bone mineral content between transplantation of PGS/BMP with or without ROB cells. These results show that transplantation of osteoblastic cells after induction of osteoblast maturation in vitro by cultivation on PGS/BMP is a potent technique for cell therapy of bone repair. PMID- 11502480 TI - Strontium ranelate inhibits bone resorption while maintaining bone formation in alveolar bone in monkeys (Macaca fascicularis). AB - Strontium ranelate (S12911) has previously been shown to stimulate bone formation and inhibit bone resorption in rats. To determine whether strontium ranelate affects normal bone remodeling, we studied the effect of strontium ranelate on alveolar bone in monkeys. Strontium ranelate, at dosages of 100, 275, and 750 mg/kg per day, or vehicle, were given by gavage to 31 normal adult monkeys (Macaca fascicularis) (15 males, 16 females), aged 3-4 years. Treatment for 6 months with strontium ranelate resulted in an increase in plasma strontium concentration. Histomorphometric analyses of indices of bone formation and resorption were determined in standardized areas of alveolar bone. Treatment with strontium ranelate decreased the histomorphometric indices of bone resorption (osteoclast surface and number) with a maximal significant effect at the highest dose tested. In contrast to this inhibitory effect on bone resorption, strontium ranelate maintained bone formation. Although the amount of osteoid tended to increase, strontium ranelate, even at the highest dose, had no deleterious effect on bone mineralization, as evaluated by mineral apposition rate and osteoid thickness. These findings show that strontium ranelate decreases indices of bone resorption while maintaining bone formation in the alveolar bone in monkeys. PMID- 11502481 TI - Fractal dimension analysis of weight-bearing bones of rats during skeletal unloading. AB - Fractal analysis was used to quantify changes in trabecular bone induced through the use of a rat tail-suspension model to simulate microgravity-induced osteopenia. Fractal dimensions were estimated from digitized radiographs obtained from tail-suspended and ambulatory rats. Fifty 4-month-old male Sprague-Dawley rats were divided into groups of 24 ambulatory (control) and 26 suspended (test) animals. Rats of both groups were killed after periods of 1, 4, and 8 weeks. Femurs and tibiae were removed and radiographed with standard intraoral films and digitized using a flatbed scanner. Square regions of interest were cropped at proximal, middle, and distal areas of each bone. Fractal dimensions were estimated from slopes of regression lines fitted to circularly averaged plots of log power vs. log spatial frequency. The results showed that the computed fractal dimensions were significantly greater for images of trabecular bones from tail suspended groups than for ambulatory groups (p < 0.01) at 1 week. Periods between 1 and 4 weeks likewise yielded significantly different estimates (p < 0.05), consistent with an increase in bone loss. In the tibiae, the proximal regions of the suspended group produced significantly greater fractal dimensions than other regions (p < 0.05), which suggests they were more susceptible to unloading. The data are consistent with other studies demonstrating osteopenia in microgravity environments and the regional response to skeletal unloading. Thus, fractal analysis could be a useful technique to evaluate the structural changes of bone. PMID- 11502482 TI - Alendronate increases degree and uniformity of mineralization in cancellous bone and decreases the porosity in cortical bone of osteoporotic women. AB - The strength of bone is correlated with bone mass but is also influenced significantly by other factors such as structural properties of the matrix (e.g., collagen mutations) and the mineral. Changes at all levels of this organization could contribute to fracture risk. We investigated the effects of alendronate (Aln) treatment on the density of mineralization and the ultrastructure of the mineral/collagen composite, size and habitus of mineral particles in iliac cancellous bone, as well as on the porosity of iliac cortical bone from postmenopausal osteoporotic women. Twenty-four transiliac bone biopsies from Phase III Aln (10 mg/day) trials (placebo and Aln after 2 and 3 years of treatment, n = 6 per group) were studied. The mineral structure was investigated by quantitative backscattered electron imaging (qBEI) and by scanning small-angle X-ray scattering (scanning-SAXS). qBEI histograms reflect the bone mineralization density distribution (BMDD), whereas SAXS patterns characterize the size and arrangement of the mineral particles in bone. We found that: (i) the relative calcium content of osteoporotic bone was significantly lower than that of data base controls; (ii) mineralization was significantly higher and more uniform after Aln treatment; (iii) size and habitus of the mineral particles was not different between placebo and Aln-treated groups; and (iv) the porosity of cortical bone was reduced significantly by Aln treatment. We conclude that Aln treatment increases the degree and uniformity of bone matrix mineralization without affecting the size and habitus of the mineral crystals. It also decreases the porosity of the corticalis. Together these effects may contribute to the observed reduction in fractures. PMID- 11502483 TI - Comparison of bone parameters by dual-energy X-ray absorptiometry and peripheral quantitative computed tomography in Hutterite vs. non-Hutterite women aged 35-60 years. AB - A previous report of elevated dual-energy X-ray absorptiometry (DXA) bone mineral density (BMD) Z scores suggests that Hutterite females might be significantly less likely to develop osteoporosis compared with other U.S. females. In the present study, we sought to determine if high Hutterite DXA BMD Z scores were elevated because of larger bone size. Hutterites reside in isolated, self sufficient colonies with an emphasis on agricultural production, and girls enter a strenuous task rotation at age 15 years. We obtained cross-sectional bone measurements of the 66% distal tibia using peripheral quantitative computed tomography (pQCT) to compare bone size and geometry on 97 Hutterite and 30 non Hutterite women, aged 35-60 years. Total body (TB) and lumbar bone mineral content (BMC), BMD, and bone area measurements by DXA were available on a subset of the study population. We identified no differences between groups in pQCT total bone area, cortical bone area, or cortical bone density. Larger bone area by DXA was apparent in Hutterites compared with non-Hutterites at the TB (least square means: 2038 +/- 8 cm2 vs. 1953 +/- 19 cm2, p < 0.05) and lumbar (least square means: 58 +/- 0.5 cm2 vs. 57 +/- 2 cm2, p < 0.01) sites. TB BMC adjusted for TB bone area was marginally higher in Hutterites compared with non-Hutterites (least square means: 2341 +/- 15 g vs. 2281 +/- 30 g, p = 0.08). Hutterites had marginally higher TB BMD Z scores when controlling for weight and age (least square means: 1.3 +/- 0.1 vs. 0.8 +/- 0.2, p = 0.07). Hutterites had higher lumbar BMC adjusted for lumbar bone area and weight (least square means: 65 +/- 1 g vs. 58 +/- 2 g, p < 0.01) and higher weight-and age-adjusted lumbar BMD Z scores (least square means: 1.1 +/- 0.1 vs. 0.1 +/- 0.4, p = 0.01). Our data indicate that a true advantage in trabecular bone density probably exists among Hutterite women aged 35-60 years. Hutterite women might be protected against age related fractures because of their larger bone size and higher bone density at normally susceptible trabecular sites. PMID- 11502484 TI - Temperature effect in the avian osteoclast resorption assay [corrected]. PMID- 11502485 TI - Alzheimer movement re-examined 25 years later: is it a "disease" or a senile condition in medical nature? AB - Dementia in the elderly used to be rare, but why has it become a major social threat today? There can be many potential answers, but an ultimate one is clear: the longer life expectancy today. This knowledge indicates that "advanced aging" is a primary suspect in the origin of senile dementia. If so, then why can many elderly remain healthy at the same old age? We know, for example, that elderly people commonly have a certain degree of atherosclerosis and osteoporosis, but only some of them develop severe clinical symptoms at the same age. These different outcomes generally can be explained by "risk factors" in life (exercise, diet, individual background, etc). It thus appears to be a general pattern that advanced aging (after age 80) will set the stage for various senile disorders, but risk factors largely determine the onset age as well as individual specificity of their clinical manifestations. In this context, senile disorders including senile dementia would differ fundamentally from the pathogen-caused conventional diseases (AIDS, polio, cancer, Down's, etc.) by origin, incidence, and intervention strategy. This view would call into question the current definition of senile dementia as a conventional "disease" (Alzheimer's). The term "Alzheimer's disease" originally referred to "midlife" dementia, but it is defined today to be the same medical entity as senile dementia on the basis that they both display the same hallmarks and symptoms despite their onset age difference. Now, after in-depth scrutiny, we finally come to realize that they are not the same disease, but as different as heart failure at midlife versus the "same" failure at advanced age (i.e., a conventional disease versus a senile condition). Thus, by eliminating the age difference, the new definition has converted a senile condition into a conventional "disease", thereby changing the course of its scientific inquiry to miss the main targets. This may be why after extensive studies for 25 years, the origin of senile dementia has remained an enigma. PMID- 11502486 TI - The role of bone marrow transplantation in first remission of paediatric all. AB - Although intensive chemotherapy has improved event-free survival for most children with lymphoblastic leukaemia there remain up to 10% who have not benefited from this approach. These include infants, children with Ph1 positive leukaemia, with near-haploidy, and slow remitters in most of whom event free survival remains below 40%. Evaluation of the benefits of Bone Marrow Transplantation in high risk ALL is fraught with difficulties and to date has not produced clear evidence of benefit. The way forward lies in prospective evaluation of BMT in tightly defined subsets of highest risk children, a task which will require international collaboration. PMID- 11502487 TI - Pediatric stem cell transplantation and critical care (an outcome evaluation). AB - This paper reviews eight published studies of children who required critical care following a stem cell transplant. Approximately 14% of children required mechanical ventilation following stem cell transplant. Sixteen percent of these children survived. Eleven percent of children who had primary lung injury secondary to either infectious or non-infectious causes survived. Patients with respiratory failure induced by disease in organ systems other than the lungs had much better survival (33-39%). Children reported to have a non-bacterial infectious lung disease had very poor survival. Children who developed multi organ system failure (MOSF) in addition to lung disease also had poor survival. The majority of children died of MOSF or pulmonary failure. PMID- 11502488 TI - Could bronchial asthma be an endogenous, pulmonary expression of retinoid intoxication? AB - Asthma has become a major public health problem, affecting about 17 million people in the United States, including 4.8 million children. A striking increase in asthma and other forms of atopy has occurred in children in the U.S. and other western countries during the past 30 years. Several studies have reported an inverse association between childhood infectious illness and the development of atopy, suggesting that certain forms of infection protect against and even inhibit asthma. This may involve a shift in the balance of CD4 T lymphocyte helper cells from a Th2 to a Th1-type cytokine profile. However, the underlying mechanisms remain uncertain. Based on a review of the literature, it is conjectured that in the absence of certain types of childhood infection, retinoids (vitamin A and its congeners) accumulate in the lung. Later, upon exposure to known triggers for asthma, retinoid metabolites may be produced in such high concentration that they produce an acute, localized form of retinoid intoxication, recognized as status asthmaticus. PMID- 11502490 TI - Ethnic diversity of class III genes in autoimmune disease. AB - One may wonder why the genes for the class III region are even situated in the human Major Histocompatibility Locus (MHC). However, on closer inspection we find that the genes for the complement components, tumor necrosis factor (TNF) and others may play an important role in host immune defenses. Thus, this region on chromosome six may be more appropriately thought of as an immune response area. Accordingly, there is a high degree of polymorphism in many genes within the MHC, the most notable being the HLA genes. In the class III region C2, factor B (Bf) and C4 are polymorphic in as many populations studied to date. The purpose of this review will be to briefly describe the class III region, identify the genetic polymorphism found in various ethnic groups and define their roles in autoimmune diseases. PMID- 11502489 TI - Molecular pathogenesis of pneumococcal pneumonia. AB - The past two decades have witnessed an explosion of data on the molecular pathogenesis of pneumonia caused by Streptococcus pneumoniae, one of the most important pathogens currently plaguing man. Identification and functional analysis of genes and their proteins, elucidation of mechanisms involved in adherence, colonization, inflammation, and invasion, and an understanding of interactions with the host and with external factors have provided knowledge that can be used to attack this organism with small molecule or vaccine based strategies. Study of the pneumococcus has also led to insights into other pathogens that share a unique spectrum of respiratory disease. In this review we will discuss recent advances in our understanding of the pathogenesis of pneumonia due to S. pneumoniae, highlighting emerging themes common to other organisms such as Haemophilus influenzae and Neisseria meningitidis. PMID- 11502491 TI - A long-term ventricular assist device utilizing a magnetic bearing system and implantable physiologic controller. PMID- 11502492 TI - Minimally invasive subclavian/axillary artery to coronary artery bypass (SAXCAB): review and classification. AB - BACKGROUND AND PURPOSE: Subclavian/axillary artery to coronary artery bypass (SAXCAB) surgery is defined as a minimally (or less) invasive coronary revascularization procedure where one or more grafts are anastomosed to the second or third parts of the subclavian artery or any of the three parts of the axillary artery (inflow source) and attached to one or more coronary arteries, and where there are two separate minimally invasive incisions to expose the target coronary artery and the inflow sources, respectively. The indications and contraindications for SAXCAB surgery are discussed, and the relevant chest wall anatomy and that of the subclavian and axillary arteries are reviewed. The effect of respiration and anatomic variability as they impact the SAXCAB graft are discussed. Three components of the anatomy that are important in SAXCAB surgery are discussed: The relation of the first rib to the clavicle insofar as it affects access to the third part of the subclavian artery, the anatomy of the subclavian and axillary arteries and their branches, and the anatomy of the chest wall and its movement. In addition, the different SAXCAB variations that have been applied clinically are reviewed and classified, and future aspects of SAXCAB research are discussed. SAXCAB surgery is unique among the different types of minimally invasive direct coronary artery bypass (MIDCAB) surgery because of the enormous diversity of the techniques that have been described. Based on these descriptions, a new classification of SAXCAB grafting is proposed depending on whether the graft is inside or outside the rib cage and whether or not the coronary artery is exposed by rib resection or through an intercostal space. The third part of the classification takes into consideration the mode of entry into the chest, whether it is by rib resection or through an intercostal space. METHODS: Inquiries were made by telephone and by mail in the year 2000 to a number of surgeons who had published details of their SAXCAB techniques, and informal information was obtained by a series of personal communications as to the estimated number of operations they had performed and the outcomes. Published data was also used to formulate a rough guide as to the international status of the procedure at this time. RESULTS: The total estimated international experience is about 100 cases and the patency is between 70 and 100 percent in the time frame of about one to two years. CONCLUSIONS: The MIDCAB technique in general has been successful in providing an alternative way to revascularize the coronary arteries, and the SAXCAB has proved to be one of the most interesting classes of MIDCAB surgery. SAXCAB grafts seem to be unique among coronary revascularization procedures and, indeed, probably almost all vascular procedures, in that there is enormous diversity in the route for the graft from the inflow source to the target coronary artery. Being knowledgeable about the different varieties of SAXCAB surgeries will help the surgeon during a rescue operation as the surgery can be tailored to suit a particular patient. The SAXCAB seems to be a very safe operation, and it is striking that so far no one has reported any major complications. PMID- 11502493 TI - Comparison of minithoracotomy with conventional sternotomy methods in valve surgery. AB - BACKGROUND: To determine the differences in the operative findings between the two groups of patients who had undergone either minithoracotomy or conventional sternotomy. METHODS: We compared 12 valve operations that were performed in our clinic with minithoracotomy (group I) between January 1997 and November 1999 with 13 valve operations that were performed with conventional median sternotomy (group II) in the same period in regard to preoperative, perioperative and postoperative variables, retrospectively. Preoperative variables were age, sex, bleeding time, clotting time, platelet count, and additional diseases like diabetes mellitus, hypertension, etc. Perioperative variables were extracorporeal circulation (ECC) time, cross-clamp (CC) time, and operation time. Postoperative variables were mechanical ventilation period, stay in the postoperative intensive care unit and hospital, mediastinal drainage amount, the amount of blood and blood products for transfusions, and costs. Group I consist of six mitral valve replacements (MVRs), three aortic valve replacements (AVRs), one aortic valve replacement combined with mitral valvuloplasty, and two tricuspid valve replacements (TVRs). Group II consist of nine MVRs and four AVRs. RESULTS: Statistical results are given with mean standard error (SEM) deviations. There were significant differences between the two groups in respect to operation time (in group I, mean operation time was 328 +/- SEM 22 minutes in group II, 271 +/- SEM 14 minutes (p < 0.04)); mediastinal drainage (in group I, mean drainage time was 283 +/- SEM 57 cc/m2, in group II, 490 +/- SEM 74 cc/m2 (p < 0.04)); and amounts of transfused blood and blood products (in group I, mean transfused blood products amount was 375 +/- SEM 115 cc/m2, in group II, 874 +/- SEM 184 cc/m2 (p < 0.03)). CONCLUSION: The operation times are apparently longer in the minithoracotomy group. On the other hand, less mediastinal drainage occurred and less blood and blood products transfusion needs were determined to exist in the minithoracotomy group. PMID- 11502494 TI - Redo mitral surgery using the Estech endoclamp. AB - BACKGROUND: Redo-CABG surgery remains extremely hazardous in the presence of open bypass grafts. In our patients with mitral valve pathology with open and well functioning bypass grafts, we explored alternative approaches in order to avoid damage to the grafts by extensive dissection and direct clamping of the ascending aorta. The "Estech procedure," which uses the Estech remote access perfusion (RAP) endoclamp catheter (Estech Inc., Danville, CA), was selected for these patients. METHODS: From January 1998 to January 2000, 10 patients underwent an Estech procedure for redo mitral surgery. All patients had previous cardiac operations such as coronary artery bypass grafting (CABG) and/or mitral valve procedures. The Estech procedure consisted of an anterior left thoracotomy and peripheral cannulation at femoral site using the Estech endovascular balloon technique. The series was comprised of seven mitral valve replacements, two valve reconstructions, and one closure of a paravalvular leak. One procedure had to be converted to a standard re-sternotomy due to extreme arteriosclerosis of the descending aorta with plaque dislocation at the time of catheter insertion. However, no damage was inflicted to the open bypass grafts. RESULTS: The follow up period ranged from six to 30 months and was 100% complete. We encountered one hospital death in our group, which was due to a late post-operative intestinal infarction and multiple organ failure (MOF), and was not procedure related. As expected, morbidity was high in this compromised cohort, but no late death has occurred prior to submission of this article. All survivors progressed to an acceptable NYHA functional class. CONCLUSION: The excellent results in this complex patient group inspired us to use the Estech procedure as a standard approach for redo mitral surgery. PMID- 11502495 TI - Correction of congenital heart defects and mitral valve operations using limited anterolateral thoracotomy. AB - PURPOSE: Median sternotomy, which generally is used as a standard access for atrial septal defect (ASD) and mitral valve operations, has a significant risk of postoperative instability/osteomyelitis of the sternum. Moreover, especially in young women, the resulting large scar is a poor cosmetic result that may have adverse psychological consequences. Our presentation suggests that these difficulties may be avoided by the use of a less invasive approach consisting of a limited anterolateral thoracotomy with standard cannulation. MATERIAL AND METHODS: From June 1997 until December 1999, 13 women, mean age 31.9 +/- 9.2 years, with atrial septum defect (n = 8), sinus venosus defect with partial anomalous pulmonary venous connection (n = 1), left atrial myxoma (n =1) or mitral valve regurgitation (n = 3), were scheduled for less invasive operation. In all cases a double lumen tube was used for ventilation. After a submammarian skin incision of about 10 cm a limited anterolateral thoracotomy was performed in the fifth right intercostal space. For cannulation of the ascending aorta a trochar cannula was used. Both caval veins were cannulated by angled vena cava catheters. Standard cardiopulmonary bypass was established using normothermia in all patients undergoing operations with correction of congenital heart defects and mild hypothermia (32 degrees C) in the three patients undergoing mitral valve operation. Surgery was performed in cardioplegic arrest using Bretschneider's solution. All corrections of congenital heart defects were performed by Goretex patches. Mitral valve reconstruction was carried out in two patients, and one patient underwent mitral valve replacement. RESULTS: No complications occurred in any of the 13 patients peri- or postoperatively. Total time of operation was 211.9 +/- 36.0 minutes, the perfusion time was 77.0 +/- 25.8 minutes, and the aortic cross-clamp time was 51.8 +/- 21.9 minutes. Mean stay in ICU was 1.2 +/- 0.4 days (total hospital stay: 7.8 +/- 2.2 days). Postoperative thoracic x-ray and cardiac echocardiography/dopplersonography revealed no pathological findings in any patients. CONCLUSION: Atrial septal defect operations, including partial anomalous pulmonary venous connection, left atrial myxoma and mitral valve operations, can be performed safely and effectively using a limited anterolateral thoracotomy and standard cannulation technique with excellent cosmetic results. PMID- 11502496 TI - Predictors of operative time in multicenter port-access valve registry: institutional differences in learning. AB - BACKGROUND: The predictors of operative time and the effects of learning in isolated valve operations using port-access techniques have not been defined. METHODS: Analysis of covariance was used to examine the determinants of procedure time, pump time, and aortic clamp time. In the largest prospective, registry of patients undergoing isolated aortic valve replacement (AVR, N=199), mitral repair (MVP, N=307), or mitral replacement (MVR, N=232) using port-access techniques 1997-1999 at 27 institutions. RESULTS: Institutional case volume ranged from one to 214 (median 6). Operative time was longer in redo procedures (5.3 +/- 1.6 vs. 4.4 +/- 1.3 hr, p = 0.0001), longer with MVP or MVR vs. AVR (4.8 +/- 1.2 vs. 5.0 +/- 1.5 vs. 3.8 +/- 1.2 hr, p = 0.0001), and decreased with case number (mean decrease 1.00 +/- 0.19 min/case, p = 0.04). Operative time also varied between institutions (p = 0.001). Rate of learning (decrease in time per case) varied significantly between institutions only for MVP (p = 0.03). Similar analysis showed that pump time and clamp times did not significantly change over time (p > 0.17) but varied significantly between institutions. Institutional volume did not affect operative, pump, or clamp times or rate of learning (decrease in operative time/case). CONCLUSIONS: These prospective registry data demonstrate that, for port-access valve procedures, procedure times continue to improve (learning) even after 100 cases. Procedure time and learning are affected by institutional differences and by the type of procedure, but are little affected by institutional volume. This data provides a model to understand learning of new surgical procedures, and this data suggests that port-access valve procedures can be mastered by a variety of institutions. PMID- 11502497 TI - Interleukin-1, interleukin-2 and interleukin-10 expression in different techniques of saphenous vein harvesting. AB - BACKGROUND: Levels of the cytokines Interleukin-1 (IL-1), IL-2, and IL-10 are sensitive to the traumatic effect of saphenous vein harvesting. Their levels are compared between the endoscopic and traditional open techniques of harvesting. METHODS: Samples of human saphenous veins were harvested from 90 randomly selected patients undergoing coronary artery bypass surgery (CABG), using the open or endoscopic techniques. Endothelial cells collected from the vein samples retrieved through both techniques were cultured for 72 hours. Pre and postoperative sera, in addition to the supernatants from the cultures, were analyzed for IL-1, IL-2, and IL-10 using ELISA. RESULTS: Mean preoperative concentrations of IL-1, Il-2, and IL-10 were 0.11+/- 0.04, 0.09 +/- 0.04, and 0.09 +/- 0.04 pg/ml, respectively. Corresponding values for postoperative sera were 0.13 +/- 0.08, 0.12 +/- 0.10, 0.14 +/- 0.17 pg/ml, respectively. The differences between pre and postoperative means for each cytokine were not statistically significant (p = 0.13, 0.18, 0.05, respectively). Mean IL-1, IL-2, and IL-10 concentrations for endothelial cell culture supernatants did not differ significantly between the endoscopic (0.17 +/- 0.11, 0.11 +/- 0.05, and 0.32 +/- 0.40 pg/ml, respectively) and the open method (0.19 +/- 0.16, 0.11 +/- 0.05, and 0.46 +/- 0.80 pg/ml, respectively) (p = 0.48, 0.81, 0.30, respectively). CONCLUSION: Since endoscopic and open saphenectomies are technically comparable with respect to their effects on IL-1, Il-2, and IL-10 levels, we recommend the endoscopic method for its lower morbidity and the potential for earlier hospital discharge. PMID- 11502498 TI - Should we use video-assisted endoscopic vein harvesting as a standard technique? AB - BACKGROUND: To investigate the feasability and results of endoscopic vein harvesting (EVH) using the Vasoview Uniport system (Guidant Corporation, Menlo Park,CA). Can this technique be used as a standard technique for vein harvesting in coronary artery bypass surgery (CABG) or is it too time consuming? Do smaller incisions result in less morbidity and discomfort? METHODS: From October 1998 to May 1999, 158 patients who underwent CABG with venous grafts, in addition to arterial grafts, formed the study population for EVH. In group A (n=131) the vein was harvested with the Vasoview Uniport System. In group B (n=27) the vein was harvested by a conventional open technique with interrupted incisions because of unavailability of the equipment. Recordings were made on vein length, harvest time, length of incision, and complications. RESULTS: In none of the patients in group A was a conversion to the open technique necessary. In 72/131, pure EVH was used. In 59/131 an additional incision below the knee was used for harvesting extra vein length. Mean harvested vein graft length (cm) was 35.9 (range 18-56) in group A and 30.6 (range 16-51) in group B, and mm of vein harvested/min was 77 and 71 in group A and B. Mean time for harvesting and closing (min) was 56.1 (range 14-120) SD 20.4 and 78.3 (range 37-129) SD 26 for a mean length of incision (cm) of six (range 2-19) and 27 (range 12-54). Wound complications at postoperative day three at discharge, and after six weeks were seen in 30 (23%), 27 (20%) and four (4%) patients of group A, and in five (18%), five (18%) and four (23%) of group B. CONCLUSIONS: Despite a learning curve in using endoscopic techniques, the total procedural time for EHV is acceptable and even shorter than open harvesting. Most of the time is gained in closure of the wound. Hematoma formation is the most common peroperative complication, but diminishes with experience. The absence of postoperative edema after EVH is striking. Despite the higher costs for disposable material, we have adopted EVH as a standard technique since patient and surgeon satisfaction have improved substantially. PMID- 11502500 TI - A multicenter initial clinical experience with right heart support and beating heart coronary surgery. AB - BACKGROUND: During coronary surgery without CPB, exposure of posterior vessel via sternotomy can cause deterioration of cardiac hemodynamics requiring inotrope drugs support. Recent animal experiments demonstrate hemodynamic benefit of right heart support (RHS) with the AMED system. The purpose of this study was to evaluate the hemodynamic effects during cardiac manipulation to expose the posterior coronary arteries, and determine the effect of RHS in restoring hemodynamics, increasing anastomotic exposure and reducing inotropic requirements. MATERIAL AND METHODS: From July 28 to December 29, 32 patients (25 men/7 women), mean age of 63.4 (+/- 6.2 years, ages: 49-78) received coronary revascularization with the A-Med RHS device. They were divided into two groups of 16 patients, A and B. Group A patients had at least one circumflex branch bypassed. The anterior wall was systematically bypassed off-pump without RHS. The right coronary artery (RCA) and the obtuse coronary artery (OM) were completed utilizing RHS. In group B patients, all vessels including anterior vessels were bypassed with the RHS. Mean arterial pressure (MAP), mean pulmonary arterial pressure (PAP), cardiac output (CO) and the average pump flow (APF) were recorded during the OM and RCA bypass for group A, and for group B LAD data was also recorded. RESULTS: Elective beating heart coronary artery bypass graft (CABG) was successfully accomplished in 32 patients with RHS. Data measurements recorded in Group A showed the improved hemodynamic recovery for OM and RCA bypass with RHS. The MAP increased from 44 to 68 mmHg (OM) and from 63 to 81 mmHg (RCA); the CO from 2.1 to 4.4 L/min (OM) and from 3.3 to 4.7 L/min (RCA). In group B, the data recorded showed the stability of the MAP in all vessels bypassed (LAD, OM and RCA). No device-related patient incidents occurred. All 32 patients were discharged to their homes. CONCLUSIONS: The AMED system, as RHS support, facilitated coronary bypass without CPB to posterior vessels, restoring hemodynamics, providing better exposure to anastomotic sites and apparently reducing inotropes need. Prospective randomize trials are necessary to confirm this initial experience. PMID- 11502501 TI - Active perfusion of coronary grafts facilitates complex off-pump coronary artery bypass surgery. AB - BACKGROUND: Hemodynamic stability during cardiac manipulation for complex, multivessel off-pump coronary artery bypass grafting (OPCAB) remains problematic. METHODS: A servo-controlled pump has been utilized to deliver warm whole blood to coronary grafts prior to construction of proximal anastomoses. RESULTS: This technique may avoid detrimental hemodynamic decompensation, which may accompany regional coronary ischemia during cardiac displacement. It may also allow precise infusion of supplemental additives leading to coronary vasodilatation, myocardial resuscitation, and enhancement of myocardial contractility. CONCLUSION: In this report, three complex OPCAB cases are described which were successfully performed with active graft perfusion and which might not otherwise have been technically feasible by conventional OPCAB techniques. PMID- 11502502 TI - Late outcome of reoperative coronary revascularization on the beating heart. AB - BACKGROUND: Reoperative (redo) coronary artery bypass grafting (CABG) is associated with a higher morbidity and mortality than first-time CABG. An off cardiopulmonary bypass (off-pump) approach to redo CABG, however, may potentially benefit redo patients. The aim of the present report is to describe the early and long-term clinical outcome of patients who underwent off-pump redo CABG between July 1985 and January 1999 in our institution. METHODS: Redo patients (n = 138) represented 13% of patients who had off-pump CABG during the period of study (n = 1072). Mean patient age was 63 +/- 12 years, and 67% were men. Surgical approaches included median sternotomy (n = 93, 67%), anterior (n = 20, 15%) and lateral (n = 25, 18%) minimally invasive direct coronary artery bypass (MIDCAB). RESULTS: Operative mortality was 2% (n = 3). Target lesion re-intervention was 6% (n = 9) Actuarial survival at a mean period of follow-up of 2.5 +/- 1 year (range: 1 month to 11 years) was 83%. Event-free survival (freedom from death, myocardial infarction, and repeat intervention) was 67%. Overall cardiac-related mortality was 10% (n = 14). CONCLUSION: Off-pump redo CABG can be safely performed with a relatively low mortality rate and a low rate of target lesion revascularization. PMID- 11502503 TI - Myocardial revascularization on the beating heart after recent onset of acute myocardial infarction. AB - OBJECTIVE: Coronary artery bypass grafting (CABG) after the recent onset of acute myocardial infarction (AMI) is associated with high morbidity and mortality. Myocardial revascularization without cardiopulmonary bypass (CPB) has been proposed as an alternative technique to treat these patients in an attempt to decrease the operative risks. METHODS: From January 1995 to June 1999, 518 patients underwent CABG after the recent onset of AMI (1-20 days): 421 patients were revascularized on-CPB and 97 patients off-CPB. Preoperative risk factors (redo operations, congestive heart failure, stroke, extensive calcification of the aorta, and dialysis) were significantly higher in the off-CPB group (p-value < 0.05). Preoperative use of intra-aortic balloon pump (IABP) (off-CPB 5.2% versus on-CPB 2.4%, p-value = NS) and emergent operations (off-CPB 5.2% versus on CPB 2.6%, p-value = NS) were similar in both groups. Mean number of grafts per patient was 3.46 in the on-CPB group versus 1.82 in the off-CPB group (p-value < 0.005). RESULTS: Actual mortality was 2.9% in the on-CPB group versus 6.2% in the off-CPB group (p-value = NS). Morbidity was comparable in the two groups. Multivariate analysis showed that advanced age, preoperative hemodynamic instability, and left ventricular hypertrophy were independent risk factors for death. Global ischemic time and preoperative hypertension were independently related to postoperative AMI. At univariate and multivariate analysis, CPB was not related to mortality or major postoperative complications. CONCLUSION: Multivariate analysis demonstrates that CABG can be performed safely with or without CPB in patients with recent AMI. CPB is not independently related to mortality or major adverse outcomes. PMID- 11502504 TI - Optimizing off-pump coronary artery bypass graft: technical and metabolic aspects. AB - PURPOSE: Our purpose was to determine if complete revascularization could be successfully performed off bypass in the majority of coronary artery bypass graft (CABG) patients. METHODS: Using a metabolic protocol and the Medtronic Octopus stabilizer device (Medtronic, Inc., Minneapolis, MN), 285 consecutive patients between July 1, 1997 and July 31, 1999 were successfully revascularized off pump via median sternotomy. A relative contraindication for off bypass CABG was cardiogenic shock. The metabolic protocol was designed to enhance systolic and diastolic cardiac function during surgery. RESULTS: Of 298 patients on whom we attempted off-pump revascularizations, 96% (285/298) were successful. The mean number of grafts per case was 3.3. The mean age of patients was 69.5 years, and 38% (109/285) were 75 years of age or older. Perioperative myocardial infarction (MI) occurred in two patients (0.7%) and operative mortality was 2.8% (8/285). Average Parsonnet score was 4.8. Cost savings were realized through minimal blood product utilization (average 1.5 units per case) and decreased cardiopulmonary bypass supplies. CONCLUSION: Complete revascularization can be performed off bypass in the majority of CABG patients. The concomitant use of the Octopus off bypass technique with aggressive metabolic support ensures manual manipulation of a beating heart and results in a low incidence of postoperative complications and significant cost savings. PMID- 11502505 TI - Closed chest coronary artery bypass on the beating heart. AB - Minimally invasive surgical procedures have become a part of routine cardiac surgery. The surgical techniques have been developed for the treatment of coronary artery disease in order to minimize surgical trauma. With the introduction of a 3-D-based totally endoscopically functioning system into minimally invasive cardiac (MIC) surgery, further reduction of skin incisions became possible and enhanced MIC techniques could be improved. Due to the 6 degrees freedom of motion allowed by wrist-enhanced instruments and a newly developed endoscopic stabilizer, totally endoscopic coronary artery bypass procedures on a beating heart became feasible. We present here our initial series of totally endoscopic "off-pump" coronary artery bypass grafting in patients suffering from coronary artery single vessel disease. In all patients, the procedure was successfully performed via four 1 cm chest incisions as closed chest procedures. PMID- 11502506 TI - Diversity of SHV and TEM beta-lactamases in Klebsiella pneumoniae: gene evolution in Northern Taiwan and two novel beta-lactamases, SHV-25 and SHV-26. AB - A total of 113 blood culture isolates of Klebsiella pneumoniae from 10 hospitals in northern Taiwan were studied for SHV and TEM beta-lactamase production. bla(SHV) was amplified from all isolates by PCR. TEM-type resistance, was found in 32 of the isolates and was of the TEM-1 type in all isolates. SHV-1, -2, -5, 11, and -12 and two novel enzymes were identified. These novel enzymes were designated SHV-25 and SHV-26 and had pIs of 7.5 and 7.6, respectively. Amino acid differences in comparison to the amino acid sequence of bla(SHV-1) were found at positions T18A (ThrACC-->AlaGCC), L35Q (LeuCTA-->GluCAA), and M129V (MetATG- >ValGTG) for SHV-25 and at position A187T (AlaGCC-->ThrACC) for SHV-26. The results of substrate profiles and MIC determinations showed that the novel enzymes did not hydrolyze extended-spectrum cephalosporins, rendering the isolates susceptible to these agents. Inhibition profiles revealed that the 50% inhibitory concentration for SHV-26 was higher than those for SHV-1 and SHV-25, resulting in an intermediate resistance to amoxicillin-clavulanic acid. Forty nine ribotypes were identified, suggesting that major clonal spread had not occurred in any of the hospitals. According to the amino acid sequence, SHV beta lactamases in Taiwan may basically be derived through stepwise mutation from SHV 1 or SHV-11 and further subdivided by four routes. The stepwise mutations initiated from SHV-1 or SHV-11 to SHV-2, SHV-5, and SHV-12 comprise the evolutionary change responsible for extended-spectrum beta-lactamase (ESBL) production in Taiwan. The stepwise mutations that lead to a non-ESBL (SHV-25) and the beta-lactamase (SHV-26) with reduced susceptibility to clavulanic acid are possibly derived from SHV-11 and SHV-1, respectively. The results suggest a stepwise evolution of SHV beta-lactamases in Taiwan. PMID- 11502507 TI - Aminoglycoside resistance with homogeneous and heterogeneous populations of antibiotic-resistant ribosomes. AB - Aminoglycosides bind to rRNA in the small subunit of the bacterial ribosome. Mutations in the decoding region of 16S rRNA confer resistance to specific subsets of aminoglycoside antibiotics. The two major classes of 2 deoxystreptamine aminoglycosides are the 4,5- and the 4,6-disubstituted antibiotics. Antibiotics of the 4,5-disubstituted class include neomycin, paromomycin, and ribostamycin. Gentamicins and kanamycins belong to the 4,6 disubstituted class of aminoglycosides. Structural studies indicated the potential importance of position 1406 (Escherichia coli numbering) in the binding of ring III of the 4,6-disubstituted class of aminoglycosides to 16S rRNA. We have introduced a U1406-to-A mutation in a plasmid-encoded copy of E. coli 16S rRNA which has been expressed either in a mixture with wild-type ribosomes or in a strain in which all rRNA is transcribed from the plasmid-encoded rrn operon. High-level resistance to many of the 4,6-disubstituted aminoglycosides is observed only when all the rRNA contains the U1406-to-A mutation. In contrast to the partial dominance of resistance observed with other mutations in the decoding region, there is a dominance of sensitivity with the 1406A mutation. Chemical footprinting experiments indicate that resistance arises from a reduced affinity of the antibiotic for the rRNA target. These results demonstrate that although position 1406 is an important determinant in the binding and action of the 4,6 disubstituted aminoglycosides, other rRNA mutations that perturb the binding of ring I of both classes of 2-deoxystreptamine aminoglycosides confer higher levels of resistance as well as a partial dominance of resistance. PMID- 11502508 TI - In vitro and in vivo activities of syn2836, syn2869, syn2903, and syn2921: new series of triazole antifungal agents. AB - The in vitro and in vivo activities of four azole compounds belonging to a new series of 2(2,4-difluorophenyl)-3-(4-substituted piperazin-1-yl)-1-(1,2,4-triazol 1-yl) butanol antifungal agents is described. The compounds were selected from a library of azole compounds synthesized by our group. The in vitro activities of Syn2869, Syn2836, Syn2903, and Syn2921 against a panel of over 240 recently collected clinical isolates of yeast and molds were determined, and the results were compared with those obtained with fluconazole (FLC), itraconazole (ITC), and amphotericin B (AMB). The MICs at which 90% of the isolates were inhibited (MIC(90)s) for the four test compounds for strains of Candida spp. ranged from <0.048 to 0.78 microg/ml. All compounds were also active against FLC-resistant Candida albicans and other Candida sp. strains. Moreover, MIC(90)s for strains of Cryptococcus neoformans, Aspergillus spp., Trichophyton spp., and Microsporum spp. were also low and ranged from <0.048 to 0.39 microg/ml. The test compounds produced a fungistatic pattern during the time-kill kinetic studies. In vivo studies indicated that all four test compounds have good efficacies against C. albicans in a murine systemic infection model and significantly improved the survival rates of the infected mice. The results for Syn2903 were similar to those for FLC, while the other compounds were slightly less effective but had ranges of activities similar to the range of activity of ITC. The compounds were also evaluated against an Aspergillus fumigatus systemic infection. Syn2903 was also superior to ITC, whereas the efficacy data for the other compounds were similar to those for ITC. It was concluded from the data generated for this new series of azole compounds in the studies described above that further pharmacokinetic and toxicologic evaluations are warranted prior to selection of a candidate compound for preclinical testing. PMID- 11502509 TI - In vitro activity of telithromycin against Spanish Streptococcus pneumoniae isolates with characterized macrolide resistance mechanisms. AB - The susceptibilities to telithromycin of 203 Streptococcus pneumoniae isolates prospectively collected during 1999 and 2000 from 14 different geographical areas in Spain were tested and compared with those to erythromycin A, clindamycin, quinupristin-dalfopristin, penicillin G, cefotaxime, and levofloxacin. Telithromycin was active against 98.9% of isolates (MICs, < or =0.5 microg/ml), with MICs at which 90% of isolates are inhibited being 0.06 microg/ml, irrespective of the resistance genotype. The corresponding values for erythromycin were 61.0% (MICs, < or =0.25 microg/ml) and >64 microg/ml. The erm(B) gene (macrolide-lincosamide-streptogramin B resistance phenotype) was detected in 36.4% (n = 74) of the isolates, which corresponded to 93.6% of erythromycin-intermediate and -resistant isolates, whereas the mef(A) gene (M phenotype [resistance to erythromycin and susceptibility to clindamycin and spiramycin without blunting]) was present in only 2.4% (n = 5) of the isolates. One of the latter isolates also carried erm(B). Interestingly, in one isolate for which the erythromycin MIC was 2 microg/ml, none of these resistance genes could be detected. Erythromycin MICs for S. pneumoniae erm(B)-positive isolates were higher (range, 0.5 to >64 microg/ml) than those for erm(B)- and mef(A)-negative isolates (range, 0.008 to 2 microg/ml). The corresponding values for telithromycin were lower for both groups, with ranges of 0.004 to 1 and 0.002 to 0.06 microg/ml, respectively. The erythromycin MIC was high for a large number of erm(B)-positive isolates, but the telithromycin MIC was low for these isolates. These results indicate the potential usefulness of telithromycin for the treatment of infections caused by erythromycin-susceptible and -resistant S. pneumoniae isolates when macrolides are indicated. PMID- 11502510 TI - Resistance of Streptococcus pneumoniae to deformylase inhibitors is due to mutations in defB. AB - Resistance to peptide deformylase inhibitors in Escherichia coli or Staphylococcus aureus is due to inactivation of transformylase activity. Knockout experiments in Streptococcus pneumoniae R6x indicate that the transformylase (fmt) and deformylase (defB) genes are essential and that a def paralog (defA) is not. Actinonin-resistant mutants of S. pneumoniae ATCC 49619 harbor mutations in defB but not in fmt. Reintroduction of the mutated defB gene into wild-type S. pneumoniae R6x recreates the resistance phenotype. The altered enzyme displays decreased sensitivity to actinonin. PMID- 11502511 TI - Pharmacokinetic and pharmacodynamic parameters for antimicrobial effects of cefotaxime and amoxicillin in an in vitro kinetic model. AB - An in vitro kinetic model was used to study the relation between pharmacokinetic and pharmacodynamic (PK-PD) parameters for antimicrobial effect, e.g., the time above MIC (T>MIC), maximum concentration in serum (C(max)), and area under the concentration-time curve (AUC). Streptococcus pyogenes and Escherichia coli were exposed to cefotaxime, and the activity of amoxicillin against four strains of Streptococcus pneumoniae with different susceptibilities to penicillin was studied. The drug elimination rate varied so that the T>MIC ranged from 20 to 100% during 24 h, while the AUC and/or the initial concentration (C(max)) were kept constant. For S. pyogenes and E. coli, the maximal antimicrobial effect (E(max)) at 24 h occurred when the antimicrobial concentration exceeded the MIC for 50 and 80% of the strains tested, respectively. The penicillin-susceptible pneumococci (MIC, 0.03 mg/liter) and the penicillin-intermediate strain (MIC, 0.25 mg/liter) showed maximal killing by amoxicillin at a T>MIC of 50%. For a strain for which the MIC was 2 mg/liter, C(max) needed to be increased to achieve the E(max). Under the condition that C(max) was 10 times the MIC, E(max) was obtained at a T>MIC of 60%, indicating that C(max), in addition to T>MIC, may be an important parameter for antimicrobial effect on moderately penicillin resistant pneumococci. For the strain for which the MIC was 4 mg/liter, the reduction of bacteria varied from -0.4 to -3.6 log(10) CFU/ml at a T>MIC of 100%, despite an initial antimicrobial concentration of 10 times the MIC. Our studies have shown that the in vitro kinetic model is a useful complement to animal models for studying the PK-PD relationship for antimicrobial effect of antibiotics. PMID- 11502512 TI - N-terminal fatty acid substitution increases the leishmanicidal activity of CA(1 7)M(2-9), a cecropin-melittin hybrid peptide. AB - In order to improve the leishmanicidal activity of the synthetic cecropin A melittin hybrid peptide CA(1-7)M(2-9) (KWKLFKKIGAVLKVL-NH(2)), a systematic study of its acylation with saturated linear fatty acids was carried out. Acylation of the N(epsilon)-7 lysine residue led to a drastic decrease in leishmanicidal activity, whereas acylation at lysine 1, in either the alpha or the epsilon NH(2) group, increased up to 3 times the activity of the peptide against promastigotes and increased up to 15 times the activity of the peptide against amastigotes. Leishmanicidal activity increased with the length of the fatty acid chain, reaching a maximum for the lauroyl analogue (12 carbons). According to the fast kinetics, dissipation of membrane potential, and parasite membrane permeability to the nucleic acid binding probe SYTOX green, the lethal mechanism was directly related to plasma membrane permeabilization. PMID- 11502513 TI - Mechanism of therapeutic effectiveness of cefixime against typhoid fever. AB - beta-Lactams have been considered ineffective against organisms growing inside mammalian cells because of their poor penetration into cells. However, cefixime has been shown to be clinically effective against typhoid fever. The probable mechanism of therapeutic effectiveness of cefixime against typhoid fever was investigated using Salmonella enterica serovar Typhimurium instead of S. enterica serovar Typhi both in a cellular and in a mouse infection model. Cefixime was able to inhibit the growth of serovar Typhimurium inhabiting monocyte-derived THP 1 cells. Elongation of serovar Typhimurium in THP-1 cells was observed microscopically. Apparent morphological changes of serovar Typhimurium in THP-1 cells were also observed by electron microscopy. The concentration of cefixime inside THP-1 cells was almost half (46 to 48%) of the concentration outside the cells when serovar Typhimurium coexisted in the solution. The length of time after oral dosing (8 mg/kg) that cefixime was present-calculated from levels in serum-at a concentration above the MIC at which 90% of the serovar Typhi organisms inside human cells were inhibited was presumed to be more than 12 h. Cefixime also showed excellent activity in the mouse systemic and oral infection models based on infections caused by serovar Typhimurium. It is concluded that a fair amount of cefixime can enter mammalian cells and inhibit the growth of bacteria inside cells when the bacteria are sensitive enough to cefixime, as are serovars Typhimurium and Typhi. PMID- 11502514 TI - In vitro and in vivo antibacterial activities of TAK-083, an agent for treatment of Helicobacter pylori infection. AB - The antibacterial activity of TAK-083 was tested against 54 clinical isolates of Helicobacter pylori and was compared with those of amoxicillin, clarithromycin, and metronidazole. The growth-inhibitory activity of TAK-083 was more potent than that of amoxicillin, clarithromycin, or metronidazole (the MICs at which 90% of the strains are inhibited were 0.031, 0.125, 64, and 8 microg/ml, respectively). The antibacterial activity of TAK-083 was highly selective against H. pylori; there was a >30-fold difference between the concentration of TAK-083 required to inhibit the growth of H. pylori and that required to inhibit the growth of common aerobic and anaerobic bacteria. Exposure of H. pylori strains to TAK-083 at the MIC or at a greater concentration resulted in an extensive loss of viability. When four H. pylori strains were successively subcultured in the medium containing subinhibitory concentrations of TAK-083, no significant change in the MICs of this compound was observed. TAK-083 strongly inhibited the formation of tryptophanyl-tRNA in H. pylori while exhibiting little effect on the same system in eukaryotes. TAK-083 was efficacious in the treatment of gastric infection caused by H. pylori in Mongolian gerbils. The results presented here indicate that TAK-083 is a promising candidate for the treatment of H. pylori infection. PMID- 11502515 TI - Continuous versus intermittent infusion of vancomycin in severe Staphylococcal infections: prospective multicenter randomized study. AB - A continuous infusion of vancomycin (CIV) may provide an alternative mode of infusion in severe hospital-acquired methicillin-resistant staphylococcal (MRS) infections. A multicenter, prospective, randomized study was designed to compare CIV (targeted plateau drug serum concentrations of 20 to 25 mg/liter) and intermittent infusions of vancomycin (IIV; targeted trough drug serum concentrations of 10 to 15 mg/liter) in 119 critically ill patients with MRS infections (bacteremic infections, 35%; pneumonia, 45%). Microbiological and clinical outcomes, safety, pharmacokinetics, ease of treatment adjustment, and cost were compared. Microbiological and clinical outcomes and safety were similar. CIV patients reached the targeted concentrations faster (36 +/- 31 versus 51 +/- 39 h, P = 0.029) and fewer samples were required for treatment monitoring than with IIV patients (7.7 +/- 2.2 versus 11.8 +/- 3.9 per treatment, P < 0.0001). The variability between patients in both the area under the serum concentration-time curve (AUC(24h)) and the daily dose given over 10 days of treatment was lower with CIV than with IIV (variances, 14,621 versus 53,975 mg(2)/liter(2)/h(2) [P = 0.026] and 414 versus 818 g(2) [P = 0.057], respectively). The 10-day treatment cost per patient was $454 +/- 137 in the IIV group and was 23% lower in the CIV group ($321 +/- 81: P < 0.0001). In summary, for comparable efficacy and tolerance, CIV may be a cost-effective alternative to IIV. PMID- 11502516 TI - Alkyl-lysophospholipid resistance in multidrug-resistant Leishmania tropica and chemosensitization by a novel P-glycoprotein-like transporter modulator. AB - Drug resistance has emerged as a major impediment in the treatment of leishmaniasis. Alkyl-lysophospholipids (ALP), originally developed as anticancer drugs, are considered to be the most promising antileishmanial agents. In order to anticipate probable clinical failure in the near future, we have investigated possible mechanisms of resistance to these drugs in Leishmania spp. The results presented here support the involvement of a member of the ATP-binding cassette (ABC) superfamily, the Leishmania P-glycoprotein-like transporter, in the resistance to ALP. (i) First, a multidrug resistance (MDR) Leishmania tropica line overexpressing a P-glycoprotein-like transporter displays significant cross resistance to the ALP miltefosine and edelfosine, with resistant indices of 9.2- and 7.1-fold, respectively. (ii) Reduced expression of P-glycoprotein in the MDR line correlates with a significant decrease in ALP resistance. (iii) The ALP were able to modulate the P-glycoprotein-mediated resistance to daunomycin in the MDR line. (iv) We have found a new inhibitor of this transporter, the sesquiterpene C 3, that completely sensitizes MDR parasites to ALP. (v) Finally, the MDR line exhibits a lower accumulation than the wild-type line of bodipy-C(5)-PC, a fluorescent analogue of phosphatidylcholine that has a structure resembling that of edelfosine. Also, C-3 significantly increases the accumulation of the fluorescent analogue to levels similar to those of wild-type parasites. The involvement of the Leishmania P-glycoprotein-like transporter in resistance to drugs used in the treatment of leishmaniasis also supports the importance of developing new specific inhibitors of this ABC transporter. PMID- 11502517 TI - Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. AB - Candida albicans is implicated in many biomaterial-related infections. Typically, these infections are associated with biofilm formation. Cells in biofilms display phenotypic traits that are dramatically different from those of their free floating planktonic counterparts and are notoriously resistant to antimicrobial agents. Consequently, biofilm-related infections are inherently difficult to treat and to fully eradicate with normal treatment regimens. Here, we report a rapid and highly reproducible microtiter-based colorimetric assay for the susceptibility testing of fungal biofilms, based on the measurement of metabolic activities of the sessile cells by using a formazan salt reduction assay. The assay was used for in vitro antifungal susceptibility testing of several C. albicans strains grown as biofilms against amphotericin B and fluconazole and the increased resistance of C. albicans biofilms against these antifungal agents was demonstrated. Because of its simplicity, compatibility with a widely available 96 well microplate platform, high throughput, and automation potential, we believe this assay represents a promising tool for the standardization of in vitro antifungal susceptibility testing of fungal biofilms. PMID- 11502518 TI - SHV-16, a beta-lactamase with a pentapeptide duplication in the omega loop. AB - A clinical isolate of Klebsiella pneumoniae was found to be resistant to ampicillin (MIC of 128 microg/ml), ticarcillin (MIC of 512 microg/ml), and ceftazidime (MIC of 128 microg/ml) and susceptible to all other beta-lactams; a synergistic effect between clavulanate and ceftazidime suggested the presence of an extended-spectrum beta-lactamase (ESBL). Transconjugants in Escherichia coli were obtained at low levels (10(-7) per donor cell) and exhibited a similar beta lactam resistance pattern (resistant to ampicillin, ticarcillin, and ceftazidime at 64 microg/ml). The ESBL, pI 7.6, was encoded by a large plasmid (>100 kb) which did not carry any other resistance determinant. The ESBL-encoding gene was amplified by PCR using bla(SHV)-specific primers and was sequenced. The deduced amino acid sequence of the SHV-16 ESBL showed that it differed from SHV-1 by only a pentapeptide insertion (163DRWET167) corresponding to a tandem duplication in the omega loop. The implication of the 163a-DRWET163b-DRWET sequence in ceftazidime resistance was confirmed by cloning either bla(SHV-1) or bla(SHV-16) in the same vector, subsequently introduced in the same E. coli strain. Under these isogenic conditions, SHV-16 conferred a 32-fold increase in ceftazidime MIC compared to that with SHV-1. Furthermore, site-directed mutagenesis experiments modifying either E166aA or E166bA revealed that the functional glutamic residue was that located in the first copy of the duplicated sequence. But surprisingly, the second E166b also conferred a low-level resistance to ceftazidime. This work is the first description of a class A enzyme exhibiting an extended substrate specificity due to an insertion instead of a nucleotide substitution(s) in a clinical isolate. PMID- 11502519 TI - In vitro activities of fluoroquinolones against the spirochete Borrelia burgdorferi. AB - Little is known to date about the in vitro activity of fluoroquinolones against Borrelia species. Our study aimed at determining the in vitro activities of 15 quinolones against nine isolates of the Borrelia burgdorferi sensu lato complex in addition to one Borrelia valaisiana and one Borrelia bissettii tick isolate. For the determination of MICs, a standardized colorimetric microdilution method was applied. Determination of minimal borreliacidal concentrations providing 100% killing of the final inoculum (MBCs) after 72 h and time-kill experiments were performed by conventional culture in Barbour-Stoenner-Kelly medium in combination with dark-field microscopy. The rank order of potency on a microgram-per milliliter basis for the substances with in vitro activity against B. burgdorferi was gemifloxacin (MIC at which 90% of the isolates tested are inhibited [MIC(90)], 0.12 microg/ml) > sitafloxacin (MIC(90), 0.5 microg/ml), grepafloxacin (MIC(90), 0.5 microg/ml) > gatifloxacin (MIC(90), 1 microg/ml), sparfloxacin (MIC(90), 1 microg/ml), trovafloxacin (MIC(90), 1 microg/ml) > moxifloxacin (MIC(90), 2 microg/ml), ciprofloxacin (MIC(90), 2 microg/ml) > levofloxacin (MIC(90), 4 microg/ml) > ofloxacin (MIC(90), 8 microg/ml), norfloxacin (MIC(90), 8 microg/ml) > fleroxacin (MIC(90), >16 microg/ml), and pefloxacin (MIC(90), 32 microg/ml) > nalidixic acid (MIC(90), 256 microg/ml). After 72 h of exposure, gemifloxacin was borreliacidal (100% killing) against the isolates investigated at a median MBC of 4 microg/ml. In the other compounds tested, median MBCs were higher (> or =8 microg/ml). Results of electron microscopy and time-kill studies clearly support an in vitro activity of some fluoroquinolones against borreliae. Our study demonstrates for the first time the enhanced in vitro effectiveness of some of the recently introduced 4-quinolones against B. burgdorferi. PMID- 11502520 TI - In vitro susceptibilities of wild-type or drug-resistant hepatitis B virus to (-) beta-D-2,6-diaminopurine dioxolane and 2'-fluoro-5-methyl-beta-L arabinofuranosyluracil. AB - Prolonged treatment of chronic hepatitis B virus (HBV) infection with lamivudine ([-]-beta-L-2',3'-dideoxy-3' thiacytidine) or famciclovir may select for viral mutants that are drug resistant due to point mutations in the polymerase gene. Determining whether such HBV mutants are sensitive to new antiviral agents is therefore important. We used a transient transfection system to compare the sensitivities of wild-type HBV and four lamivudine- and/or famciclovir-resistant HBV mutants to adefovir [9-(2-phosphonyl-methoxyethyl)-adenine; PMEA] and the nucleoside analogues (-)-beta-D-2, 6-diaminopurine dioxolane (DAPD) and 2'-fluoro 5-methyl-beta-L-arabinofuranosyluracil (L-FMAU). The drug-resistant mutants contained amino acid substitutions in the polymerase protein. We found that the M550I and M550V plus L526M substitutions, which confer lamivudine resistance, did not confer cross-resistance to adefovir or DAPD, but conferred cross-resistance to L-FMAU. The M550V substitution in isolation conferred a similar phenotype to M550I, except that it did not confer significant resistance to L-FMAU. The L526M substitution, which is associated with famciclovir resistance, conferred cross resistance to L-FMAU but not to adefovir or DAPD. Inhibition of HBV secretion by DAPD, L-FMAU, and adefovir did not always correlate with inhibition of the generation of intracellular HBV replicative intermediates, suggesting that these analogs may preferentially inhibit specific stages of the viral replication cycle. PMID- 11502521 TI - Occurrence and risk of cochleotoxicity in cystic fibrosis patients receiving repeated high-dose aminoglycoside therapy. AB - Cystic fibrosis (CF) patients receive repeated courses of aminoglycoside therapy. These patients would consequently be expected to be more susceptible to cochleotoxicity, a recognized side effect with single courses of aminoglycoside therapy. The primary aim of this retrospective study was to establish the incidence and severity of auditory deficit in CF patients. Standard (0.25- to 8 kHz) and high-frequency (10- to 16-kHz) pure-tone audiometry was carried out in 70 CF patients, and the results were compared with the results from 91 control subjects. These subjects were further divided into pediatric and adult groups. Of 70 CF patients, 12 (1 pediatric) displayed hearing loss considered to be caused by repeated exposure to aminoglycosides. There was a nonlinear relationship between the courses of therapy received and the incidence of hearing loss. The severity of the loss did not appear to be related to the number of courses received. Assuming the risk of loss to be independent for each course, preliminary estimates of per course risk of hearing loss were less than 2%. Upon comparison with previous clinical studies and experimental work, these findings suggest that the incidence of cochleotoxicity in CF patients is considerably lower than would be expected, suggesting that the CF condition may confer protection against aminoglycoside cochleotoxicity. PMID- 11502522 TI - Specific inhibition of human immunodeficiency virus type 1 (HIV-1) integration in cell culture: putative inhibitors of HIV-1 integrase. AB - To study the effect of potential human immunodeficiency virus type 1 (HIV-1) integrase inhibitors during virus replication in cell culture, we used a modified nested Alu-PCR assay to quantify integrated HIV DNA in combination with the quantitative analysis of extrachromosomal HIV DNA. The two diketo acid integrase inhibitors (L-708,906 and L-731,988) blocked the accumulation of integrated HIV-1 DNA in T cells following infection but did not alter levels of newly synthesized extrachromosomal HIV DNA. In contrast, we demonstrated that L17 (a member of the bisaroyl hydrazine family of integrase inhibitors) and AR177 (an oligonucleotide inhibitor) blocked the HIV replication cycle at, or prior to, reverse transcription, although both drugs inhibited integrase activity in cell-free assays. Quercetin dihydrate (a flavone) was shown to not have any antiviral activity in our system despite reported anti-integration properties in cell-free assays. This refined Alu-PCR assay for HIV provirus is a useful tool for screening anti-integration compounds identified in biochemical assays for their ability to inhibit the accumulation of integrated HIV DNA in cell culture, and it may be useful for studying the effects of these inhibitors in clinical trials. PMID- 11502523 TI - Isolation of rat dihydrofolate reductase gene and characterization of recombinant enzyme. AB - While assays of many antifolate inhibitors for dihydrofolate reductase (DHFR) have been performed using rat DHFR as a target, neither the sequence nor the structure of rat DHFR is known. Here, we report the isolation of the rat DHFR gene through screening of a rat liver cDNA library. The rat liver DHFR gene has an open reading frame of 561 bp encoding a protein of 187 amino acids. Comparisons of the rat enzyme with those from other species indicate a high level of conservation at the primary sequence level and more so for the amino acid residues comprising the active site of the enzyme. Expression of the rat DHFR gene in bacteria produced a recombinant protein with high enzymatic activity. The recombinant protein also paralleled the human enzyme with respect to the inhibition by most of the antifolates tested with PT652 and PT653 showing a reversal in their patterns. Our results indicated that rat DHFR can be used as a model to study antifolate compounds as potential drug candidates. However, variations between rat and human DHFR enzymes, coupled with unique features in the inhibitors, could lead to the observed differences in enzyme sensitivity and selectivity. PMID- 11502524 TI - In vitro activities of 10 antifungal drugs against 508 dermatophyte strains. AB - We have tested 508 strains belonging to 24 species of dermatophytes against 10 antifungal drugs following mainly the NCCLS (M38-P) standard for filamentous fungi. However, several important factors, such as the temperature (28 versus 35 degrees C) and time of incubation (4 to 10 days versus 21 to 74 h), have been modified. The antifungals used were itraconazole, ketoconazole, miconazole, clotrimazole, voriconazole, terbinafine, amphotericin B, fluconazole, UR-9825, and G-1. In general, with the exception of fluconazole and G-1, all antifungals were shown to be highly effective. PMID- 11502525 TI - Pharmacokinetics of clinafloxacin after single and multiple doses. AB - Clinafloxacin (CI-960) is a potent broad-spectrum, fluoroquinolone antibiotic that has been studied for parenteral and oral administration in patients with serious infections. The objectives of these studies were to examine the pharmacokinetics and safety of clinafloxacin following administration of single and twice-daily intravenous (i.v.) and oral doses to volunteers. Plasma and urine samples were assayed by validated liquid chromatographic methods, and pharmacokinetic parameter values were determined by noncompartmental methods. Safety was evaluated by clinical observation and laboratory tests. Absorption was rapid after oral administration, with maximum concentrations in plasma (C(max)) generally occurring within 2 h. Concentrations in plasma declined biexponentially, with an average terminal half-life of 4 to 6 h after single doses and 5 to 7 h after multiple doses. Increases in C(max) and area under the concentration-time curves (AUC) were generally proportional to the dose. The volume of distribution was much greater than total body water. Approximately 40 to 75% of the clinafloxacin doses were excreted unchanged into urine. Absolute bioavailability of orally administered clinafloxacin was approximately 90% and did not change with increasing dose. Therefore, switching patients from i.v. to oral dosing should achieve similar concentrations in plasma. The tolerability of clinafloxacin was acceptable. No serious adverse events occurred. C(max) values and minimum plasma clinafloxacin concentrations during multiple dosing exceeded MICs for a wide range of organisms. PMID- 11502526 TI - Clinafloxacin pharmacokinetics in subjects with various degrees of renal function. AB - As the primary route for elimination of clinafloxacin is renal clearance (CL(R)) of unchanged drug, studies were conducted to determine the pharmacokinetic profile of clinafloxacin following administration to young and elderly subjects, subjects with various degrees of renal function, and subjects requiring dialysis. These were open-label studies in which subjects received single oral clinafloxacin doses. Sixteen young subjects (18 to 35 years old) and 16 elderly subjects (>65 years old) were enrolled in a study comparing pharmacokinetic profiles of clinafloxacin in young and elderly subjects. Twenty subjects having various degrees of renal function were enrolled into one of three groups based on degree of renal function as measured by creatinine clearance (CL(CR)). Twelve subjects with severe renal impairment requiring dialysis enrolled in a third study. Clinafloxacin was generally well tolerated by all subjects. Clinafloxacin pharmacokinetic profiles in elderly subjects were dependent only on age-related decreases in renal function. Clinafloxacin maximum concentrations in plasma, areas under the concentration-time curves, and terminal elimination half-life values increased with decreasing CL(CR) values. Total apparent body clearance of clinafloxacin from the plasma after oral administration (CL(oral)) and CL(R) were dependent on CL(CR) according to the following relationships: CL(oral) = 2.3. CL(CR) + 77 and CL(R) = 1.74. CL(CR). Hemodialysis had no significant effect on clinafloxacin clearance. Based on the relationship between CL(CR) and clinafloxacin CL(oral) and CL(R) values, the clinafloxacin dose should be halved in patients having a CL(CR) of <40 ml/min. Further dose adjustment is not warranted in patients requiring hemodialysis. PMID- 11502527 TI - Drug interactions with clinafloxacin. AB - Many fluoroquinolone antibiotics are inhibitors of cytochrome P450 enzyme systems and may produce potentially important drug interactions when administered with other drugs. Studies were conducted to determine the effect of clinafloxacin on the pharmacokinetics of theophylline, caffeine, warfarin, and phenytoin, as well as the effect of phenytoin on the pharmacokinetics of clinafloxacin. Concomitant administration of 200 or 400 mg of clinafloxacin reduces mean theophylline clearance by approximately 50 and 70%, respectively, and reduces mean caffeine clearance by 84%. (R)-Warfarin concentrations in plasma during clinafloxacin administration are 32% higher and (S)-warfarin concentrations do not change during clinafloxacin treatment. An observed late pharmacodynamic effect was most likely due to gut flora changes. Phenytoin has no effect on clinafloxacin pharmacokinetics, while phenytoin clearance is 15% lower during clinafloxacin administration. PMID- 11502529 TI - Expression of ubiquitin gene in Microsporum canis and Trichophyton mentagrophytes cultured with fluconazole. AB - The expression of the ubiquitin (Ub) gene in dermatophytes was examined for its relation to resistance against the antifungal drug fluconazole. The nucleotide sequences and the deduced amino acid sequences of the Ub gene in Microsporum canis were proven to be 99% similar to those of the Ub gene in Trichophyton mentagrophytes. Expression of mRNA of Ub in M. canis and T. mentagrophytes was enhanced when the fungi were cultured with fluconazole. The antifungal activity of fluconazole against these dermatophytes was increased in the presence of Ub proteasome inhibitor. PMID- 11502528 TI - Limonene arrests parasite development and inhibits isoprenylation of proteins in Plasmodium falciparum. AB - Isoprenylation is an essential protein modification in eukaryotic cells. Herein, we report that in Plasmodium falciparum, a number of proteins were labeled upon incubation of intraerythrocytic forms with either [(3)H]farnesyl pyrophosphate or [(3)H]geranylgeranyl pyrophosphate. By thin-layer chromatography, we showed that attached isoprenoids are partially modified to dolichol and other, uncharacterized, residues, confirming active isoprenoid metabolism in this parasite. Incubation of blood-stage P. falciparum treated with the isoprenylation inhibitor limonene significantly decreased the parasites' progression from the ring stage to the trophozoite stage and at 1.22 mM, 50% of the parasites died after the first cycle. Using Ras- and Rap-specific monoclonal antibodies, putative Rap and Ras proteins of P. falciparum were immunoprecipitated. Upon treatment with 0.5 mM limonene, isoprenylation of these proteins was significantly decreased, possibly explaining the observed arrest of parasite development. PMID- 11502530 TI - Novel alpha- and beta-amino acid inhibitors of influenza virus neuraminidase. AB - In an effort to discover novel, noncarbohydrate inhibitors of influenza virus neuraminidase we hypothesized that compounds which contain positively charged amino groups in an appropriate position to interact with the Asp 152 or Tyr 406 side chains might be bound tightly by the enzyme. Testing of 300 alpha- and beta amino acids led to the discovery of two novel neuraminidase inhibitors, a phenylglycine and a pyrrolidine, which exhibited K(i) values in the 50 microM range versus influenza virus A/N2/Tokyo/3/67 neuraminidase but which exhibited weaker activity against influenza virus B/Memphis/3/89 neuraminidase. Limited optimization of the pyrrolidine series resulted in a compound which was about 24 fold more potent than 2-deoxy-2,3-dehydro-N-acetylneuraminic acid in an anti influenza cell culture assay using A/N2/Victoria/3/75 virus. X-ray structural studies of A/N9 neuraminidase-inhibitor complexes revealed that both classes of inhibitors induced the Glu 278 side chain to undergo a small conformational change, but these compounds did not show time-dependent inhibition. Crystallography also established that the alpha-amino group of the phenylglycine formed hydrogen bonds to the Asp 152 carboxylate as expected. Likewise, the beta amino group of the pyrrolidine forms an interaction with the Tyr 406 hydroxyl group and represents the first compound known to make an interaction with this absolutely conserved residue. Phenylglycine and pyrrolidine analogs in which the alpha- or beta-amino groups were replaced with hydroxyl groups were 365- and 2,600-fold weaker inhibitors, respectively. These results underscore the importance of the amino group interactions with the Asp 152 and Tyr 406 side chains and have implications for anti-influenza drug design. PMID- 11502531 TI - Virtual screening of combinatorial libraries across a gene family: in search of inhibitors of Giardia lamblia guanine phosphoribosyltransferase. AB - Parasitic protozoa lack the ability to synthesize purine nucleotides de novo, relying instead on purine salvage enzymes for their survival. Guanine phosphoribosyltransferase (GPRT) from the protozoan parasite Giardia lamblia is a potential target for rational antiparasitic drug design, based on the experimental evidence, which indicates the lack of interconversion between adenine and guanine nucleotide pools. The present study is a continuation of our efforts to use three-dimensional structures of parasitic phosphoribosyltransferases (PRTs) to design novel antiparasitic agents. Two micromolar phthalimide-based GPRT inhibitors were identified by screening the in house phthalimide library. A combination of structure-based scaffold selection using virtual library screening across the PRT gene family and solid phase library synthesis led to identification of smaller (molecular weight, <300) ligands with moderate to low specificity for GPRT; the best inhibitors, GP3 and GP5, had K(i) values in the 23 to 25 microM range. These results represent significant progress toward the goal of designing potent inhibitors of purine salvage in Giardia parasites. As a second step in this process, altering the phthalimide moiety to optimize interactions in the guanine-binding pocket of GPRT is expected to lead to compounds with promising activity against G. lamblia PRT. PMID- 11502532 TI - New class of small nonpeptidyl compounds blocks Plasmodium falciparum development in vitro by inhibiting plasmepsins. AB - Malarial parasites rely on aspartic proteases called plasmepsins to digest hemoglobin during the intraerythrocytic stage. Plasmepsins from Plasmodium falciparum and Plasmodium vivax have been cloned and expressed for a variety of structural and enzymatic studies. Recombinant plasmepsins possess kinetic similarity to the native enzymes, indicating their suitability for target-based antimalarial drug development. We developed an automated assay of P. falciparum plasmepsin II and P. vivax plasmepsin to quickly screen compounds in the Walter Reed chemical database. A low-molecular-mass (346 Da) diphenylurea derivative (WR268961) was found to inhibit plasmepsins with a K(i) of 1 to 6 microM. This compound appears to be selective for plasmepsin, since it is a poor inhibitor of the human aspartic protease cathepsin D (K(i) greater than 280 microM). WR268961 inhibited the growth of P. falciparum strains W2 and D6, with 50% inhibitory concentrations ranging from 0.03 to 0.16 microg/ml, but was much less toxic to mammalian cells. The Walter Reed chemical database contains over 1,500 compounds with a diphenylurea core structure, 9 of which inhibit the plasmepsins, with K(i) values ranging from 0.05 to 0.68 microM. These nine compounds show specificity for the plasmepsins over human cathepsin D, but they are poor inhibitors of P. falciparum growth in vitro. Computational docking experiments indicate how diphenylurea compounds bind to the plasmepsin active site and inhibit the enzyme. PMID- 11502533 TI - Efficacies of ABT-773, a new ketolide, against experimental bacterial infections. AB - ABT-773 is a novel ketolide effective against antibacterial-resistant respiratory tract pathogens. The pharmacokinetic profile of ABT-773 was studied in rats and consisted of a mean peak concentration in plasma of 1.07 microg/ml and an area under the concentration-time curve (AUC) of 12.03 microg. h/ml when the compound was delivered at a dose of 25 mg/kg of body weight. It concentrated in rat lung tissue, with a lung tissue-to-plasma ratio of 29 based on the AUC. In acute systemic infections in mice, ABT-773 showed efficacy against macrolide susceptible strains of Staphylococcus aureus, Streptococcus pneumoniae, S. pyogenes, and Listeria monocytogenes. Additionally, ABT-773 improved the survival of mice infected with resistant S. pneumoniae containing either the ermB gene, the mefE gene, or altered penicillin binding protein genes. In a rat lung model of infection, ABT-773 demonstrated 50% effective doses lower than those of comparator macrolides when evaluated against the following strains of S. pneumoniae: a macrolide-lincosamide-streptogramin B-susceptible strain, an ermB strain, and an mefE strain. ABT-773 was also effective against Haemophilus influenzae lung infections in rats. Thus, ABT-773 may prove to be a useful new antibacterial agent for the treatment of respiratory tract infections. PMID- 11502534 TI - Role of penicillin-binding protein 5 in expression of ampicillin resistance and peptidoglycan structure in Enterococcus faecium. AB - The contribution of penicillin-binding protein 5 (PBP 5) to intrinsic and acquired beta-lactam resistance was investigated by constructing isogenic strains of Enterococcus faecium producing different PBP 5. The pbp5 genes from three E. faecium clinical isolates (BM4107, D344, and H80721) were cloned into the shuttle vector pAT392 and introduced into E. faecium D344S, a spontaneous derivative of E. faecium D344 highly susceptible to ampicillin due to deletion of pbp5 (MIC, 0.03 microg/ml). Immunodetection of PBP5 indicated that cloning of the pbp5 genes into pAT392 resulted in moderate overproduction of PBP 5 in comparison to wild type strains. This difference may be attributed to a difference in gene copy number. Expression of the pbp5 genes from BM4107 (MIC, 2 microg/ml), D344 (MIC, 24 microg/ml), and H80721 (MIC, 512 microg/ml) in D344S conferred relatively low levels of resistance to ampicillin (MICs, 6, 12, and 20 microg/ml, respectively). A methionine-to-alanine substitution was introduced at position 485 of the BM4107 PBP 5 by site-directed mutagenesis. In contrast to previous hypotheses based on comparison of nonisogenic strains, this substitution resulted in only a 2.5-fold increase in the ampicillin MIC. The reversed-phase high-performance liquid chromatography muropeptide profiles of D344 and D344S were similar, indicating that deletion of pbp5 was not associated with a detectable defect in cell wall synthesis. These results indicate that pbp5 is a nonessential gene responsible for intrinsic resistance to moderate levels of ampicillin and by itself cannot confer high-level resistance. PMID- 11502535 TI - GES-2, a class A beta-lactamase from Pseudomonas aeruginosa with increased hydrolysis of imipenem. AB - Pseudomonas aeruginosa GW-1 was isolated in 2000 in South Africa from blood cultures of a 38-year-old female who developed nosocomial pneumonia. This isolate harbored a self-transferable ca. 100-kb plasmid that conferred an expanded spectrum cephalosporin resistance profile associated with an intermediate susceptibility to imipenem. A beta-lactamase gene, bla(GES-2), was cloned from whole-cell DNA of P. aeruginosa GW-1 and expressed in Escherichia coli. GES-2, with a pI value of 5.8, hydrolyzed expanded-spectrum cephalosporins, and its substrate profile was extended to include imipenem compared to that of GES-1, identified previously in Klebsiella pneumoniae. GES-2 activity was less inhibited by clavulanic acid, tazobactam and imipenem than GES-1. The GES-2 amino acid sequence differs from that of GES-1 by a glycine-to-asparagine substitution in position 170 located in the omega loop of Ambler class A enzymes. This amino acid change may explain the extension of the substrate profile of the plasmid-encoded beta-lactamase GES-2. PMID- 11502536 TI - Susceptibilities of Mycoplasma hominis, M. pneumoniae, and Ureaplasma urealyticum to GAR-936, dalfopristin, dirithromycin, evernimicin, gatifloxacin, linezolid, moxifloxacin, quinupristin-dalfopristin, and telithromycin compared to their susceptibilities to reference macrolides, tetracyclines, and quinolones. AB - The susceptibilities of Mycoplasma hominis, Mycoplasma pneumoniae, and Ureaplasma urealyticum to eight new antimicrobial agents were determined by agar dilution. M. pneumoniae was susceptible to the new glycylcycline GAR-936 at 0.12 microg/ml and evernimicin at 4 microg/ml, but it was resistant to linezolid. It was most susceptible to dirithromycin, quinupristin-dalfopristin, telithromycin, reference macrolides, and josamycin. M. hominis was susceptible to linezolid, evernimicin, and GAR-936. It was resistant to macrolides and the ketolide telithromycin but susceptible to quinupristin-dalfopristin and josamycin. U. urealyticum was susceptible to evernimicin (8 to 16 microg/ml) and resistant to linezolid. It was less susceptible to GAR-936 (4.0 microg/ml) than to tetracycline (0.5 microg/ml). Telithromycin and quinupristin-dalfopristin were the most active agents against ureaplasmas (0.06 microg/ml). The new quinolone gatifloxacin was active against M. pneumoniae and M. hominis at 0.12 to 0.25 microg/ml and active against ureaplasmas at 1.0 microg/ml. The MICs of macrolides were markedly affected by pH, with an 8- to 32-fold increase in the susceptibility of M. pneumoniae as the pH increased from 6.9 to 7.8. A similar increase in susceptibility with increasing pH was also observed with ureaplasmas. Tetracyclines showed a fourfold increase of activity as the pH decreased 1 U, whereas GAR-936 showed a fourfold decrease in activity with a decrease in pH. PMID- 11502537 TI - DNA sequence analysis of rdxA and frxA from 12 pairs of metronidazole-sensitive and -resistant clinical Helicobacter pylori isolates. AB - We previously reported that inactivation of rdxA and/or frxA converted Helicobacter pylori from metronidazole sensitive to metronidazole resistant. To examine the individual roles of rdxA and frxA in the development of metronidazole resistance in H. pylori, we examined the status of rdxA and frxA from 12 pairs of metronidazole-sensitive and -resistant H. pylori isolates obtained following unsuccessful therapy containing metronidazole. Arbitrary primed fingerprinting analyses revealed that the genotypes of 11 sensitive and resistant pairs of strains were essentially identical. Amino acid sequence identities of RdxA and FrxA from the 14 metronidazole-sensitive isolates ranged from 92 to 98% and 95 to 98%, respectively, compared to that of H. pylori J99 (MIC, 1 microg/ml). All strains with high-level metronidazole resistance (MICs, 128 microg/ml) contained premature truncation of both RdxA and FrxA caused by nonsense and/or frameshift mutations. Strains with intermediate resistance to metronidazole (MICs, 32 to 64 microg/ml) contained a single premature truncation and/or altered RdxA and FrxA caused by nonsense, frameshift, and unique missense mutations. The low-level metronidazole-resistant strains (MICs, 8 microg/ml) contained unique missense mutations in FrxA but no specific changes in RdxA. The results demonstrate that alterations in both the rdxA and frxA genes are required for moderate and high level metronidazole resistance and that metronidazole resistance that develops during anti-H. pylori therapy containing metronidazole is most likely to involve a single sensitive strain infection rather than a coinfection with a metronidazole-resistant strain. PMID- 11502538 TI - Cell-based fluorescence assay for human immunodeficiency virus type 1 protease activity. AB - The human immunodeficiency virus type 1 (HIV-1) protease is essential for production of infectious virus and is therefore a major target for the development of drugs against AIDS. Cellular proteins are also cleaved by the protease, which explains its cytotoxic activity and the consequent failure to establish convenient cell-based protease assays. We have exploited this toxicity to develop a new protease assay that relies on transient expression of an artificial protease precursor harboring the green fluorescent protein (GFP-PR). The precursor is activated in vivo by autocatalytic cleavage, resulting in rapid elimination of protease-expressing cells. Treatment with therapeutic doses of HIV 1 protease inhibitors results in a dose-dependent accumulation of the fluorescent precursor that can be easily detected and quantified by flow cytometric and fluorimetric assays. The precursor provides a convenient and noninfectious model for high-throughput screenings of substances that can interfere with the activity of the protease in living cells. PMID- 11502539 TI - Treatment of visceral leishmaniasis with sterically stabilized liposomes containing camptothecin. AB - The efficacy of 20(S)-camptothecin (CPT), free and incorporated into sterically stabilized liposomes, has been investigated in vitro against Leishmania donovani promastigotes and in vivo in a murine model of visceral leishmaniasis. Incubation of L. donovani promastigotes with free or liposomal CPT inhibited the growth of parasites in a dose-dependent manner. Tissue distribution studies revealed that the intraperitoneal administration of liposomal CPT was efficient for the delivery of high drug levels to the liver and spleen. Treatment of infected mice with intraperitoneal injections of free and liposomal CPT significantly reduced the parasite loads in the livers by 43 and 55%, respectively, compared with the loads for untreated controls. However, both treatments caused normochromic anemia and neutropenia. PMID- 11502540 TI - Risk factors for emergence of resistance to broad-spectrum cephalosporins among Enterobacter spp. AB - Among 477 patients with susceptible Enterobacter spp., 49 subsequently harbored third-generation cephalosporin-resistant Enterobacter spp. Broad-spectrum cephalosporins were independent risk factors for resistance (relative risk [OR] = 2.3, P = 0.01); quinolone therapy was protective (OR = 0.4, P = 0.03). There were trends toward decreased risk for resistance among patients receiving broad spectrum cephalosporins and either aminoglycosides or imipenem. Of the patients receiving broad-spectrum cephalosporins, 19% developed resistance. PMID- 11502541 TI - Interspecies recombination contributes minimally to fluoroquinolone resistance in Streptococcus pneumoniae. AB - Analysis of 71 ciprofloxacin-resistant (MIC > or = 4 microg/ml) Streptococcus pneumoniae clinical isolates revealed only 1 for which the quinolone resistance determining regions of the parC, parE, and gyrB genes were genetically related to those of viridans group streptococci. Our findings support the occurrence of interspecies recombination of type II topoisomerase genes; however, its contribution to the emergence of quinolone resistance among pneumococci appears to have been minimal. PMID- 11502542 TI - In vitro antifungal activities of the new triazole UR-9825 against clinically important filamentous fungi. AB - We used a modified reference microdilution method (the M-38P method) to evaluate the in vitro activities of the new triazole UR-9825 in comparison with those of amphotericin B against 77 strains of opportunistic filamentous fungi. UR-9825 was clearly more active than amphotericin B against all fungi except Fusarium solani and Scytalidium spp. Notably, UR-9825 had low MICs for Aspergillus fumigatus and Paecilomyces lilacinus (MICs at which 90% of isolates are inhibited, 0.125 microg/ml for both species). PMID- 11502543 TI - Pretreatment of mice with clindamycin improves survival of endotoxic shock by modulating the release of inflammatory cytokines. AB - Suppression of endotoxin release and subsequent production of inflammatory cytokines is crucial in the treatment of septic shock. We investigated the effect of clindamycin (CLI) on endotoxic shock induced in mice by Escherichia coli lipopolysaccharide (LPS). Mice were treated with CLI (160 to 600 mg/kg) or saline and then injected with E. coli LPS and D-(+)-galactosamine intraperitoneally 0.5 h after CLI administration. Pretreatment with CLI significantly improved survival in a dose-dependent manner (CLI, at 160, 300, and 440 mg/kg) and significantly lowered the peak concentrations of tumor necrosis factor alpha and interleukin 1beta (IL-1beta) in serum. However, the peak concentrations of IL-6 in the sera of CLI-treated mice were higher than in control mice. Our findings suggest that CLI alters LPS-induced inflammatory cytokine production and suppresses endotoxin induced mortality in this murine model. PMID- 11502544 TI - Stability and compatibility of ceftazidime administered by continuous infusion to intensive care patients. AB - The stability and compatibility of ceftazidime have been examined in the context of its potential use in concentrated solutions for continuous infusion in patients suffering from severe nosocomial pneumonia and receiving other intravenous medications by the same route. Ceftazidime stability in 4 to 12% solutions was found satisfactory (<10% degradation) for 24 h if kept at a temperature of 25 degrees C (77 degrees F) maximum. Studies mimicking the simultaneous administration of ceftazidime and other drugs as done in clinics showed physical incompatibilities with vancomycin, nicardipine, midazolam, and propofol and a chemical incompatibility with N-acetylcystein. Concentrated solutions (50 mg/ml) of erythromycin or clarithromycin caused the appearance of a precipitate, whereas gentamicin, tobramycin, amikacin, isepamicin, fluconazole, ketamine, sufentanil, valproic acid, furosemide, uradipil, and a standard amino acid solution were physically and chemically compatible. PMID- 11502545 TI - Emergence of a pneumococcal clone with cephalosporin resistance and penicillin susceptibility. AB - We report two South African serotype 6B pneumococcal isolates with cephalosporin resistance, yet with susceptibility to penicillin. DNA fingerprinting revealed that they were clonal in origin. pbp 2X and 1A genes showed major alterations typical of cephalosporin-resistant pneumococci. The pbp 2B gene was completely unaltered, explaining the penicillin susceptibility of the isolates. PMID- 11502546 TI - Multidrug-resistant strains of Neisseria gonorrhoeae in Greece. AB - Eighty-seven out of 575 gonococci isolated in Greece from 1991 to 1998 belonged to serovar Bropyst and exhibited resistance to penicillin, tetracycline, erythromycin, and chloramphenicol. Conventional and molecular typing showed three clusters, A, B, and C, that were associated with networks of high- frequency transmitters (cluster A with homosexuals and clusters B and C with refugees from Eastern Europe). Study of one isolate revealed mutations in the penA, mtrR, and porB genes that may explain the multidrug-resistant phenotype. PMID- 11502547 TI - Structural analysis of chloroquine resistance reversal by imipramine analogs. AB - For imipramine, desipramine, and eight analogs of these well-known drugs, an N-5 aminoalkyl substitution was a minimum but insufficient structural feature associated with chloroquine resistance reversal. Although a second distal aliphatic nitrogen atom was unnecessary for resistance reversal, the direction of the dipole moment vector was critical. PMID- 11502548 TI - Integrons and gene cassettes in the enterobacteriaceae. AB - Integrons were detected in 59 of 120 (49%) urinary isolates of Enterobacteriaceae by PCR using degenerate primers targeted to conserved regions of class 1, 2, and 3 integrase genes. PCR sequencing analysis of the cassette arrays revealed a predominance of cassettes that confer resistance to the aminoglycosides and trimethoprim. PMID- 11502549 TI - Interactions of quinupristin-dalfopristin with eight other antibiotics as measured by time-kill studies with 10 strains of Staphylococcus aureus for which quinupristin-dalfopristin alone was not bactericidal. AB - Quinupristin-dalfopristin (Q-D) and eight other antimicrobial agents were tested alone and in combination with Q-D in time-kill studies against 10 strains of macrolide-lincosamide-streptogramin B-resistant Staphylococcus aureus. Although Q D is normally a bactericidal drug, it was only bacteriostatic for these isolates. Gentamicin alone was bactericidal against 7 of the 10 strains, and Q-D did not alter that killing effect. However, when vancomycin, cefepime, ceftazidime, imipenem, piperacillin-tazobactam, and ciprofloxacin were bactericidal when tested alone, the killing rates were reduced when combined with Q-D. The clinical significance of this in vitro antagonism is unknown at this time, and more studies are needed. PMID- 11502550 TI - Escherichia coli as an expression system for K(+) transport systems from plants. AB - The value of the Escherichia coli expression system has long been established because of its effectiveness in characterizing the structure and function of exogenously expressed proteins. When eukaryotic membrane proteins are functionally expressed in E. coli, this organism can serve as an alternative to eukaryotic host cells. A few examples have been reported of functional expression of animal and plant membrane proteins in E. coli. This mini-review describes the following findings: 1) homologous K(+) transporters exist in prokaryotic cells and in eukaryotic cells; 2) plant K(+) transporters can functionally complement mutant K(+) transporter genes in E. coli; and 3) membrane structures of plant K(+) transporters can be elucidated in an E. coli system. These experimental findings suggest the possibility of utilizing the E. coli bacterium as an expression system for other eukaryotic membrane transport proteins. PMID- 11502551 TI - Ca(2+) influx and cAMP elevation overcame botulinum toxin A but not tetanus toxin inhibition of insulin exocytosis. AB - Previous reports showed that cleavage of vesicle-associated membrane protein-2 (VAMP-2) and synaptosomal-associated protein of 25 kDa (SNAP-25) by clostridial neurotoxins in permeabilized insulin-secreting beta-cells inhibited Ca(2+)-evoked insulin secretion. In these reports, the soluble N-ethylmaleimide-sensitive factor attachment protein target receptor proteins might have formed complexes, which preclude full accessibility of the putative sites for neurotoxin cleavage. In this work, VAMP-2 and SNAP-25 were effectively cleaved before they formed toxin-insensitive complexes by transient transfection of insulinoma HIT or INS-1 cells with tetanus toxin (TeTx) or botulinum neurotoxin A (BoNT/A), as shown by immunoblotting and immunofluorescence microscopy. This resulted in an inhibition of Ca(2+) (glucose or KCl)-evoked insulin release proportionate to the transfection efficiency (40-50%) and an accumulation of insulin granules. With the use of patch-clamp capacitance measurements, Ca(2+)-evoked exocytosis by membrane depolarization to -10 mV was abolished by TeTx (6% of control) but only moderately inhibited by BoNT/A (30% of control). Depolarization to 0 mV to maximize Ca(2+) influx partially overcame BoNT/A (50% of control) but not TeTx inhibition. Of note, cAMP activation potentiated Ca(2+)-evoked secretion by 129% in control cells but only 55% in BoNT/A-transfected cells and had negligible effects in TeTx-transfected cells. These results indicate that, whereas VAMP-2 is absolutely necessary for insulin exocytosis, the effects of SNAP-25 depletion on exocytosis, perhaps on insulin granule pool priming or mobilization steps, could be partially reversed by higher levels of Ca(2+) or cAMP potentiation. PMID- 11502552 TI - Zinc status affects p53, gadd45, and c-fos expression and caspase-3 activity in human bronchial epithelial cells. AB - This study was designed to examine the influence of zinc depletion and supplementation on the expression of p53 gene, target genes of p53, and caspase-3 activity in normal human bronchial epithelial (NHBE) cells. A serum-free, low zinc medium containing 0.4 micromol/l of zinc [zinc deficient (ZD)] was used to deplete cellular zinc over one passage. In addition, cells were cultured for one passage in media containing 4.0 micromol/l of zinc [zinc normal (ZN)], which represents normal culture concentrations (Clonetics); 16 micromol/l of zinc [zinc adequate (ZA)], which represents normal human plasma zinc levels; or 32 micromol/l of zinc [zinc supplemented (ZS)], which represents the high end of plasma zinc levels attainable by oral supplementation in humans. Compared with ZN cells, cellular zinc levels were 76% lower in ZD cells but 3.5-fold and 6-fold higher in ZA and ZS cells, respectively. Abundances of p53 mRNA and nuclear p53 protein were elevated in treatment groups compared with controls (ZN). For p53 mRNA abundance, the highest increase (3-fold) was observed in ZD cells. In contrast, the highest increase (17-fold) in p53 nuclear protein levels was detected in ZS cells. Moreover, gadd45 mRNA abundance was moderately elevated in ZD and ZA cells and was not altered in ZS cells compared with ZN cells. Furthermore, the only alteration in c-fos mRNA and caspase-3 activity was the twofold increase and the 25% reduction, respectively, detected in ZS compared with ZN cells. Thus p53, gadd45, and c-fos and caspase-3 activity appeared to be modulated by cellular zinc status in NHBE cells. PMID- 11502553 TI - Preservation of metabolic reserves and function after storage of myocytes in hypothermic UW solution. AB - Isolated rat myocytes cold stored anaerobically up to 24 h in University of Wisconsin solution lost 95% of their ATP and 100% of their glycogen. They underwent contracture when rewarmed in a Krebs-Henseleit (KH) medium that contained Ca unless Ca addition was delayed. In the latter case, cell function, measured by stimulation-induced cell shortening, was surprisingly well retained. Aerobically stored cells were resistant to Ca on rewarming, although 96% of glycogen was still lost, along with 46% of ATP. Cells that were incubated for 48 h aerobically with the substrates glucose and pyruvate at pH 6.2 retained 77% of their ATP and 59% of their glycogen, with good cell morphology. At pH 6.2, the demand for ATP was only 55% of that at pH 7.4. However, after rewarming, these cells functioned no better than anaerobically stored cells, although their inotropic response to isoproterenol was improved. We conclude that 1) aerobic conditions with substrates at low pH preserve myocyte metabolic reserves well for 48 h, partly by reducing the demand for ATP; 2) rewarming conditions are critical for anaerobically stored cells with metabolic stores that are severely depleted; and 3) unloaded cell function is surprisingly insensitive to a period of severe metabolic deprivation. PMID- 11502554 TI - S-adenosyl-L-homocysteine hydrolase is necessary for aldosterone-induced activity of epithelial Na(+) channels. AB - The A6 cell line was used to study the role of S-adenosyl-L-homocysteine hydrolase (SAHHase) in the aldosterone-induced activation of the epithelial Na(+) channel (ENaC). Because aldosterone increases methylation of several different molecules, and because this methylation is associated with increased Na(+) reabsorption, we tested the hypothesis that aldosterone increases the expression and activity of SAHHase protein. The rationale for this work is that general methylation may be promoted by activation of SAHHase, the only enzyme known to metabolize SAH, a potent end-product inhibitor of methylation. Although aldosterone increased SAHHase activity, steroid did not affect SAHHase expression. Antisense SAHHase oligonucleotide decreased SAHHase expression and activity. Moreover, this oligonucleotide, as well as a pharmacological inhibitor of SAHHase, decreased aldosterone-induced activity of ENaC via a decrease in ENaC open probability. The kinetics of ENaC in cells treated with antisense plus aldosterone were similar to those reported previously for the channel in the absence of steroid. This is the first report showing that active SAHHase, in part, increases ENaC open probability by reducing the transition rate from open states in response to aldosterone. Thus aldosterone-induced SAHHase activity plays a critical role in shifting ENaC from a gating mode with short open and closed times to one with longer open and closed times. PMID- 11502555 TI - Mechanism of thiamine uptake by human jejunal brush-border membrane vesicles. AB - Thiamine, a water-soluble vitamin, is essential for normal cellular functions, growth and development. Thiamine deficiency leads to significant clinical problems and occurs under a variety of conditions. To date, however, little is known about the mechanism of thiamine absorption in the native human small intestine. The objective of this study was, therefore, to characterize the mechanism of thiamine transport across the brush-border membrane (BBM) of human small intestine. With the use of purified BBM vesicles (BBMV) isolated from the jejunum of organ donors, thiamine uptake was found to be 1) independent of Na(+) but markedly stimulated by an outwardly directed H(+) gradient (pH 5.5(in)/pH 7.5(out)); 2) competitively inhibited by the cation transport inhibitor amiloride (inhibitor constant of 0.12 mM); 3) sensitive to temperature and osmolarity of the incubation medium; 4) significantly inhibited by thiamine structural analogs (amprolium, oxythiamine, and pyrithiamine), but not by unrelated organic cations (tetraethylammonium, N-methylnicotinamide, or choline); 5) not affected by the addition of ATP to the inside and outside of the BBMV; 6) potential insensitive; and 7) saturable as a function of thiamine concentration with an apparent Michaelis-Menten constant of 0.61 +/- 0.08 microM and a maximal velocity of 1.00 +/- 0.47 pmol. mg protein(-1). 10 s(-1). Carrier-mediated thiamine uptake was also found in BBMV of human ileum. These data demonstrate the existence of a Na(+)-independent, pH-dependent, amiloride-sensitive, electroneutral carrier mediated mechanism for thiamine absorption in native human small intestinal BBMV. PMID- 11502556 TI - Involvement of DMT1 in uptake of Cd in MDCK cells: role of protein kinase C. AB - The involvement of iron (Fe) transporters in the uptake of cadmium (Cd) was examined in Madin-Darby kidney cells (MDCK). The uptake of Cd displayed properties that are associated with the Fe transporter divalent metal transporter 1 (DMT1). For example, the uptake of Cd and Fe was reduced by altering the cell membrane potential. The uptake of Cd was blocked by Fe, and the uptake of Fe was blocked by Cd. Also, the uptake of Cd and Fe was higher in MDCK cells bathed in a buffer at low pH. Increased uptake of Fe and Cd was observed in the HEK-293 cell line overexpressing DMT1. Overnight treatment of MDCK cells with the protein kinase C activator phorbol 12,13-dibutyrate (PDBu) resulted in increased uptake of Cd and Fe and an increase in DMT1 mRNA. An increase in newly transcribed DMT1 mRNA was not observed, suggesting that PDBu does not increase DMT1 mRNA by activating transcription. Rather, the increase was most likely due to greater stability of DMT1 mRNA, because the rate of degradation of DMT1 mRNA was slower in MDCK cells treated with PDBu. Our results suggest that Fe and Cd are transported in MDCK cells by a transporter with biochemical properties similar to those of DMT1. PMID- 11502557 TI - ANG II type 1 receptor downregulation does not require receptor endocytosis or G protein coupling. AB - ANG II type 1 (AT(1)) receptors respond to sustained exposure to ANG II by undergoing downregulation of absolute receptor numbers. It has been assumed previously that downregulation involves endocytosis. The present study hypothesized that AT(1) receptor downregulation occurs independently of receptor endocytosis or G protein coupling. Mutant AT(1) receptors with carboxy-terminal deletions internalized <5% of radioligand compared with 65% for wild-type AT(1) receptors. The truncated AT(1) receptors retained the ability to undergo downregulation. These data suggest the existence of an alternative pathway to AT(1) receptor degradation that does not require endocytosis, per se. Point mutations in either the second transmembrane region or second intracellular loop impaired G protein (G(q)) coupling. These receptors exhibited a biphasic pattern of downregulation. The earliest phase of downregulation (0-2 h) was independent of coupling to G(q), but no additional downregulation was observed after 2 h of ANG II exposure in the receptors with impaired coupling to G(q). These data suggest that coupling to G(q) is required for the later phase (2-24 h) of AT(1) receptor downregulation. PMID- 11502558 TI - CFTR modulates programmed cell death by decreasing intracellular pH in Chinese hamster lung fibroblasts. AB - To study the potential influence of cystic fibrosis conductance regulator (CFTR) on intracellular pH regulation during apoptosis induction, we used PS120 Chinese hamster lung fibroblasts devoid of the Na(+)/H(+) exchanger (NHE1 isoform) transfected with constructs, allowing the expression of CFTR and/or NHE1. Kinetics of lovastatin-induced apoptosis were measured by orcein staining, double staining with Hoechst-33258, propidium iodide, DNA fragmentation, and annexin V labeling. In PS120 control cells, the percentage of apoptotic cells after 40 h of lovastatin treatment was 23 +/- 3%, whereas in PS120 CFTR-transfected cells, this percentage was 40 +/- 4%. In PS120 NHE1 cells, the transfection with CFTR did not modify the percentage of apoptotic cells after 40 h (control: 19 +/- 3%, n = 8; CFTR: 17 +/- 1%, n = 8), indicating that blocking intracellular acidification by overexpressing the Na(+)/H(+) exchanger inhibited the enhancement of apoptosis induced by CFTR. In all cell lines, the initial pH values were identical (pH = 7.46 +/- 0.04, n = 9), and treatment with lovastatin led to intracellular acidification. However, the pH value after 40 h was lower in PS120 CFTR transfected cells (pH = 6.85 +/- 0.02, n = 10) than in PS120 cells (pH = 7.15 +/- 0.03, n = 10). To further investigate the origin of this increased intracellular acidification observed in CFTR-transfected cells, the activity of the DIDS inhibitable Cl(-)/HCO exchanger was studied. 8-Bromoadenosine 3',5'-cyclic monophosphate incubation resulted in Cl(-)/HCO exchanger activation in PS120 CFTR transfected cells but had no effect on PS120 cells. Together, our results suggest that CFTR can enhance apoptosis in Chinese hamster lung fibroblasts, probably due to the modulation of the Cl(-)/HCO exchanger, resulting in a more efficient intracellular acidification. PMID- 11502559 TI - Direct estimate of 1:1 stoichiometry of K(+)-Cl(-) cotransport in rabbit erythrocytes. AB - This work was undertaken to obtain a direct measure of the stoichiometry of Na(+) independent K(+)-Cl(-) cotransport (KCC), with rabbit red blood cells as a model system. To determine whether (86)Rb(+) can be used quantitatively as a tracer for KCC, (86)Rb(+) and K(+) effluxes were measured in parallel after activation of KCC with N-ethylmaleimide (NEM). The rate constant for NEM-stimulated K(+) efflux into isosmotic NaCl was smaller than that for (86)Rb(+) by a factor of 0.68 +/- 0.11 (SD, n = 5). This correction factor was used in all other experiments to calculate the K(+) efflux from the measured (86)Rb(+) efflux. To minimize interference from the anion exchanger, extracellular Cl(-) was replaced with SO, and 4,4'-diisothiocyanothiocyanatodihydrostilbene-2,2'-disulfonic acid was present in the flux media. The membrane potential was clamped near 0 mV with the protonophore 2,4-dinitrophenol. The Cl(-) efflux at 25 degrees C under these conditions is approximately 100,000-fold smaller than the uninhibited Cl(-)/Cl(-) exchange flux and is stimulated approximately 2-fold by NEM. The NEM-stimulated (36)Cl(-) flux is inhibited by okadaic acid and calyculin A, as expected for KCC. The ratio of the NEM-stimulated K(+) to Cl(-) efflux is 1.12 +/- 0.26 (SD, n = 5). We conclude that K(+)-Cl(-) cotransport in rabbit red blood cells has a stoichiometry of 1:1. PMID- 11502560 TI - Phosphate-induced chondrocyte apoptosis is linked to nitric oxide generation. AB - An elevation in inorganic phosphate (P(i)) concentration activates epiphyseal chondrocyte apoptosis. To determine the mechanism of apoptosis, tibial chondrocytes were treated with P(i), and nitrate/nitrite (NO/NO) levels were determined. P(i) induced a threefold increase in the NO/NO concentration; inhibitors of nitric oxide (NO) synthase activity and P(i) transport significantly reduced NO/NO levels and prevented cell death. Furthermore, a dose dependent increase in cell death was observed after exposure of chondrocytes to S nitrosoglutathione. P(i) increased caspase 3 activity 2.7-fold. Both caspase 1 and caspase 3 inhibitors protected chondrocytes from P(i)-induced apoptosis. P(i) caused a significant decrease in the mitochondrial membrane potential, while NO synthase inhibitors maintained mitochondrial function. While P(i) caused thiol depletion, inhibition of P(i) uptake or NO generation served to maintain glutathione levels. The results suggest that NO serves to mediate key metabolic events linked to P(i)-dependent chondrocyte apoptosis. PMID- 11502561 TI - Hypertonicity rescues T cells from suppression by trauma-induced anti inflammatory mediators. AB - Trauma causes the release of anti-inflammatory factors thought to cause infections by inhibiting T cells. We have found that hypertonic saline (HS) enhances functions of normal T cells. Here we studied if HS can rescue T cells from suppression by costimulating interleukin (IL)-2 production. Human peripheral blood mononuclear cells were treated with the immunosuppressive factors IL-4, IL 10, transforming growth factor (TGF)-beta(1), and PGE(2) and with serum of trauma patients and stimulated with phytohemagglutinin, and IL-2 production was measured. Costimulation with HS tripled IL-2 production of normal cells. IL-4, IL 10, TGF-beta(1), and PGE(2) suppressed IL-2 production with IC(50) of 500, 1, 36,000, and 0.01 pg/ml, respectively. Costimulation of suppressed cells with HS restored IL-2 production and increased IC(50) values >70-fold. Serum from trauma patients could completely suppress normal cells; however, costimulation with HS restored IL-2 production by up to 80% of the control response. These findings show that HS can restore the function of suppressed T cells, suggesting that HS resuscitation of trauma patients could reduce posttraumatic sepsis. PMID- 11502562 TI - Gene and protein expressions of nitric oxide synthases in ischemia-reperfused peripheral nerve of the rat. AB - This study examined mRNA and protein expressions of neuronal (nNOS), inducible (iNOS), and endothelial nitric oxide synthases (eNOS) in peripheral nerve after ischemia-reperfusion (I/R). Sixty-six rats were divided into the ischemia only and I/R groups. One sciatic nerve of each animal was used as the experimental side and the opposite untreated nerve as the control. mRNA levels in the nerve were quantitatively measured by competitive PCR, and protein was determined by Western blotting and immunohistochemical staining. The results showed that, after ischemia (2 h), both nNOS and eNOS protein expressions decreased. After I/R (2 h of ischemia followed by 3 h of reperfusion), expression of both nNOS and eNOS mRNA and protein decreased further. In contrast, iNOS mRNA significantly increased after ischemia and was further upregulated (14-fold) after I/R, while iNOS protein was not detected. The results reveal the dynamic expression of individual NOS isoforms during the course of I/R injury. An understanding of this modulation on a cellular and molecular level may lead to understanding the mechanisms of I/R injury and to methods of ameliorating peripheral nerve injury. PMID- 11502563 TI - Regulation of ATP-sensitive K(+) channels by protein kinase C in murine colonic myocytes. AB - We investigated the regulation of ATP-sensitive K(+) (K(ATP)) currents in murine colonic myocytes with patch-clamp techniques. Pinacidil (10(-5) M) activated inward currents in the presence of high external K(+) (90 mM) at a holding potential of -80 mV in dialyzed cells. Glibenclamide (10(-5) M) suppressed pinacidil-activated current. Phorbol 12,13-dibutyrate (PDBu; 2 x 10(-7) M) inhibited pinacidil-activated current. 4-alpha-Phorbol ester (5 x 10(-7) M), an inactive form of PDBu, had no effect on pinacidil-activated current. In cell attached patches, the open probability of K(ATP) channels was increased by pinacidil, and PDBu suppressed openings of K(ATP) channels. When cells were pretreated with chelerythrine (10(-6) M) or calphostin C (10(-7) M), inhibition of the pinacidil-activated whole cell currents by PDBu was significantly reduced. In cells studied with the perforated patch technique, PDBu also inhibited pinacidil-activated current, and this inhibition was reduced by chelerythrine (10(-6) M). Acetylcholine (ACh; 10(-5) M) inhibited pinacidil-activated currents, and preincubation of cells with calphostin C (10(-7) M) decreased the effect of ACh. Cells dialyzed with protein kinase C epsilon-isoform (PKCepsilon) antibody had normal responses to pinacidil, but the effects of PDBu and ACh on K(ATP) were blocked in these cells. Immunofluorescence and Western blots showed expression of PKCepsilon in intact muscles and isolated smooth muscle cells of the murine proximal colon. These data suggest that PKC regulates K(ATP) in colonic muscle cells and that the effects of ACh on K(ATP) are largely mediated by PKC. PKCepsilon appears to be the major isozyme that regulates K(ATP) in murine colonic myocytes. PMID- 11502564 TI - Timolol may inhibit aqueous humor secretion by cAMP-independent action on ciliary epithelial cells. AB - The beta-adrenergic antagonist timolol reduces ciliary epithelial secretion in glaucomatous patients. Whether inhibition is mediated by reducing cAMP is unknown. Elemental composition of rabbit ciliary epithelium was studied by electron probe X-ray microanalysis. Volume of cultured bovine pigmented ciliary epithelial (PE) cells was measured by electronic cell sizing; Ca(2+) activity and pH were monitored with fura 2 and 2',7'-bis(2-carboxyethyl)-5(6) carboxyfluorescein, respectively. Timolol (10 microM) produced similar K and Cl losses from ciliary epithelia in HCO/CO(2) solution but had no effect in HCO/CO(2)-free solution or in HCO/CO(2) solution containing the carbonic anhydrase inhibitor acetazolamide. Inhibition of Na(+)/H(+) exchange by dimethylamiloride in HCO/CO(2) solution reduced Cl and K comparably to timolol. cAMP did not reverse timolol's effects. Timolol (100 nM, 10 microM) and levobunolol (10 microM) produced cAMP-independent inhibition of the regulatory volume increase (RVI) in PE cells and increased intracellular Ca(2+) and pH. Increasing Ca(2+) with ionomycin also blocked the RVI. The results document a previously unrecognized cAMP-independent transport effect of timolol. Inhibition of Cl(-)/HCO exchange may mediate timolol's inhibition of aqueous humor formation. PMID- 11502565 TI - Regulation of cytoplasmic calcium levels by two nitric oxide receptors. AB - We examined the effects of dissolved nitric oxide (NO) gas on cytoplasmic calcium levels ([Ca(2+)](i)) in C6 glioma cells under anoxic conditions. The maximum elevation (27 +/- 3 nM) of [Ca(2+)](i) was reached at 10 microM NO. A second application of NO was ineffective if the first was >0.5 microM. The NO donor diethylamine/NO mimicked the effects of NO. Acute exposure of the cells to low calcium levels was without effect on the NO-evoked response. Thapsigargin (TG) increased [Ca(2+)](i) and was less effective if cells were pretreated with NO. Hemoglobin inhibited the effects of NO at a molar ratio of 10:1. 8-Bromo-cGMP was without effect on the NO-evoked response. If cells were pretreated with TG or exposed chronically to nominal amounts of calcium, NO decreased [Ca(2+)](i). The results suggest that C6 glioma cells have two receptors for NO. One receptor (NO(A)) elevates [Ca(2+)](i) and resides on the endoplasmic reticulum (ER). The other receptor (NO(B)) decreases [Ca(2+)](i) and resides on the plasmalemma or the ER. The latter receptor dominates when the level of calcium within intracellular stores is diminished. PMID- 11502566 TI - EGF receptor function is required in late G(1) for cell cycle progression induced by bombesin and bradykinin. AB - We examined the role of epidermal growth factor (EGF) receptor (EGFR) tyrosine kinase activation in G protein-coupled receptor (GPCR) agonist-induced mitogenesis in Swiss 3T3 and Rat-1 cells. Addition of EGFR tyrosine kinase inhibitors (e.g., tyrphostin AG-1478) abrogated bombesin-induced extracellular signal-regulated kinase (ERK) activation in Rat-1 cells but not in Swiss 3T3 cells, indicating the importance of cell context in determining the role of EGFR in ERK activation. In striking contrast, treatment with tyrphostin AG-1478 markedly (~70%) inhibited DNA synthesis induced by bombesin in both Swiss 3T3 and Rat-1 cells. Similar inhibition of bombesin-induced DNA synthesis in Swiss 3T3 cells was obtained using four structurally different inhibitors of EGFR tyrosine kinase. Furthermore, kinetic analysis indicates that EGFR function is necessary for bombesin-induced mitogenesis in mid-late G(1) in both Swiss 3T3 and Rat-1 cells. Our results indicate that EGFR kinase activity is necessary in mid-late G(1) for promoting the accumulation of cyclins D1 and E and implicate EGFR function in the coupling of GPCR signaling to the activation of the cell cycle. PMID- 11502567 TI - High-glucose-induced metallothionein expression in endothelial cells: an endothelin-mediated mechanism. AB - Vascular endothelial cells are constantly exposed to oxidative stress and must be protected by physiological responses. In diabetes mellitus, endothelial cell permeability is impaired and may be increased by high extracellular glucose concentrations. It has been postulated that metallothionein (MT) can protect endothelial cells from oxidative stress with its increased expression by cytokines, thrombin, and endothelin (ET)-1. In this study, we demonstrate that high glucose concentration can induce MT expression in endothelial cells through a distinct ET-dependent pathway. Exposure of human umbilical vein endothelial cells (HUVEC) to increasing concentrations of glucose resulted in a rapid dose dependent increase in MT-2 and ET-1 mRNA expression. MT expression may be further augmented with addition of ET-1. Preincubation of the cells with the specific ET(B) antagonist BQ-788 blocked MT-2 mRNA expression more effectively than the ET(A) inhibitor TBC-11251. High glucose also increased immunoreactive MT protein expression and induced translocation of MT into the perinuclear area. Perinuclear localization of MT was related to high-glucose-induced reorganization of F-actin filaments. These results demonstrate that an increase in extracellular glucose in HUVEC can lead to a rapid dose-dependent increase in MT-2 mRNA expression and to perinuclear localization of MT protein with changes to the cytoskeleton. These effects are mediated via the ET receptor-dependent pathway. PMID- 11502568 TI - Proton leak and CFTR in regulation of Golgi pH in respiratory epithelial cells. AB - Work addressing whether cystic fibrosis transmembrane conductance regulator (CFTR) plays a role in regulating organelle pH has remained inconclusive. We engineered a pH-sensitive excitation ratiometric green fluorescent protein (pHERP) and targeted it to the Golgi with sialyltransferase (ST). As determined by ratiometric imaging of cells expressing ST-pHERP, Golgi pH (pH(G)) of HeLa cells was 6.4, while pH(G) of mutant (DeltaF508) and wild-type CFTR-expressing (WT-CFTR) respiratory epithelia were 6.7-7.0. Comparison of genetically matched DeltaF508 and WT-CFTR cells showed that the absence of CFTR statistically increased Golgi acidity by 0.2 pH units, though this small difference was unlikely to be physiologically important. Golgi pH was maintained by a H(+) vacuolar (V)-ATPase countered by a H(+) leak, which was unaffected by CFTR. To estimate Golgi proton permeability (P(H(+))), we modeled transient changes in pH(G) induced by inhibiting the V-ATPase and by acidifying the cytosol. This analysis required knowing Golgi buffer capacity, which was pH dependent. Our in vivo estimate is that Golgi P(H(+)) = 7.5 x 10(-4) cm/s when pH(G) = 6.5, and surprisingly, P(H(+)) decreased as pH(G) decreased. PMID- 11502569 TI - An inwardly rectifying K(+) channel, Kir4.1, expressed in astrocytes surrounds synapses and blood vessels in brain. AB - Glial cells express inwardly rectifying K(+) (Kir) channels, which play a critical role in the buffering of extracellular K(+). Kir4.1 is the only Kir channel so far shown to be expressed in brain glial cells. We examined the distribution of Kir4.1 in rat brain with a specific antibody. The Kir4.1 immunostaining distributed broadly but not diffusely in the brain. It was strong in some regions such as the glomerular layer of the olfactory bulb, the Bergmann glia in the cerebellum, the ependyma, and pia mater, while little activity was detected in white matter of the corpus callosum or cerebellar peduncle. In the olfactory bulb, Kir4.1 immunoreactivity was detected in a scattered manner in about one-half of the glial fibrillary acidic protein-positive astrocytes. Immunoelectron microscopic examination revealed that Kir4.1 channels were enriched on the processes of astrocytes wrapping synapses and blood vessels. These data suggest that Kir4.1 is expressed in a limited population of brain astrocytes and may play a specific role in the glial K(+)-buffering action. PMID- 11502570 TI - Disruption of the actin cytoskeleton regulates cytokine-induced iNOS expression. AB - Interleukin-1beta (IL-1beta) induces the inducible nitric oxide synthase (iNOS), resulting in the release of nitric oxide (NO) from glomerular mesangial cells. In this study, we demonstrated that disruption of F-actin formation by sequestration of G-actin with the toxin latrunculin B (LatB) dramatically potentiated IL-1beta induced iNOS protein expression in a dose-dependent manner. LatB by itself had little or no effect on iNOS expression. Staining of F-actin with nitrobenzoxadiazole (NBD)-phallacidin demonstrated that LatB significantly impaired F-actin stress fiber formation. Jasplakinolide (Jasp), which binds to and stabilizes F-actin, suppressed iNOS expression enhanced by LatB. These data strongly suggest that actin cytoskeletal dynamics regulates IL-1beta-induced iNOS expression. We demonstrated that LatB decreases serum response factor (SRF) activity as determined by reporter gene assays, whereas Jasp increases SRF activity. The negative correlation between SRF activity and iNOS expression suggests a negative regulatory role for SRF in iNOS expression. Overexpression of a dominant negative mutant of SRF increases the IL-1beta-induced iNOS expression, providing direct evidence that SRF inhibits iNOS expression. PMID- 11502571 TI - Polyamine depletion stabilizes p53 resulting in inhibition of normal intestinal epithelial cell proliferation. AB - The p53 nuclear phosphoprotein plays a critical role in transcriptional regulation of target genes involved in growth arrest and apoptosis. The natural polyamines, including spermidine, spermine, and their precursor putrescine, are required for cell proliferation, and decreasing cellular polyamines inhibits growth of the small intestinal mucosa. In the current study, we investigated the mechanisms of regulation of p53 gene expression by cellular polyamines and further determined the role of the gene product in the process of growth inhibition after polyamine depletion. Studies were conducted both in vivo and in vitro using rats and the IEC-6 cell line, derived from rat small intestinal crypt cells. Levels for p53 mRNA and protein, transcription and posttranscription of the p53 gene, and cell growth were examined. Depletion of cellular polyamines by treatment with alpha-difluoromethylornithine (DFMO) increased p53 gene expression and caused growth inhibition in the intact small intestinal mucosa and the cultured cells. Polyamine depletion dramatically increased the stability of p53 mRNA as measured by the mRNA half-life but had no effect on p53 gene transcription in IEC-6 cells. Induction of p53 mRNA levels in DFMO-treated cells was paralleled by an increase in the rate of newly synthesized p53 protein. The stability of p53 protein was also increased after polyamine depletion, which was associated with a decrease in Mdm2 expression. When polyamine-deficient cells were exposed to exogenous spermidine, a decrease in p53 gene expression preceded an increase in cellular DNA synthesis. Inhibition of the p53 gene expression by using p53 antisense oligodeoxyribonucleotides significantly promoted cell growth in the presence of DFMO. These findings indicate that polyamines downregulate p53 gene expression posttranscriptionally and that growth inhibition of small intestinal mucosa after polyamine depletion is mediated, at least partially, through the activation of p53 gene. PMID- 11502572 TI - Cloning and characterization of a functional P2X receptor from larval bullfrog skin. AB - ATP activates an apical-to-basolateral nonselective cation current across the skin of larval bullfrogs (Rana catesbeiana) with similarities to currents carried by some P2X receptors. A functional P2X receptor was cloned from tadpole skin RNA that encodes a 409-amino acid protein with highest protein homology to cP2X(8). RT-PCR showed that this transcript was found in skin, heart, eye, brain, and skeletal muscle of tadpoles but not in skin, brain, or heart of adults. After transcribed RNA from this clone was injected into Xenopus oocytes, application of ATP activated a transient current similar to other P2X receptors and the ATP activated transient in short-circuit current (I(sc)) across intact skin. The agonists 2-methylthio-ATP and adenosine-5'-O-(thiotriphoshate) also activated transient currents. alpha,beta-Methylene-ATP and ADP were poor agonists of this receptor. Suramin and pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid tetrasodium (PPADS) were potent antagonists, and PPADS showed an irreversible blockade of this receptor to agonist activation. Under external Na(+)-free, Ca(2+)/Mg(2+)-free conditions (N-methyl-D-glucamine replacement, 0.5 mM EGTA), ATP activated a steadily increasing inward current. Fluorescence microscopy showed that propidium was entering the cells, suggesting that a relatively large pore size was formed under zero divalent conditions. This clone has some characteristics consistent with previously described ATP-activated I(sc) in the tadpole skin. Because the clone is not found in adult skin, it may have some exclusive role in the tadpole such as sensory reception by the skin or triggering apoptosis at metamorphosis. PMID- 11502573 TI - Na(+)-dependent neutral amino acid transporter ATB(0) is a rabbit epithelial cell brush-border protein. AB - System B(0) activity accounts for the majority of intestinal and kidney luminal neutral amino acid absorption. An amino acid transport system, called ATB(0) (also known as ASCT2), with functional characteristics similar to those of system B(0), has been recently cloned. We generated polyclonal antibodies to human and rabbit ATB(0) COOH-terminal peptides and used Western blot analysis to detect ATB(0) protein in rabbit tissues, rabbit ileal brush-border membrane vesicles (BBMV), and HeLa cells transfected with plasmids containing ATB(0) cDNAs. Immunohistochemistry was used to localize ATB(0) in rabbit kidney and intestine. In Western blots of rabbit tissues, ATB(0) was a broad smear of 78- to 85-kDa proteins. In transfected HeLa cells, ATB(0) appeared as a smear consisting of 57- to 65-kDa proteins. The highest expression was found in the kidney. ATB(0) was enriched in rabbit ileal BBMV and in HeLa cells transfected with ATB(0) cDNAs. In the kidney and in the intestine, ATB(0) was confined to the brush-border membrane (BBM) of the proximal tubular cell and of the enterocyte, respectively. Tissue and intracellular distribution of ATB(0) protein parallels that of system B(0) activity. ATB(0) protein could be the transporter responsible for system B(0) in the BBM of epithelial cells. PMID- 11502574 TI - Ca(2+) regulation of gap junctional coupling in lens epithelial cells. AB - The quantitative effects of Ca(2+) signaling on gap junctional coupling in lens epithelial cells have been determined using either the spread of Mn(2+) that is imaged by its ability to quench the fluorescence of fura 2 or the spread of the fluorescent dye Alexa Fluor 594. Gap junctional coupling was unaffected by a mechanically stimulated cell-to-cell Ca(2+) wave. Furthermore, when cytosolic Ca(2+) concentration (Ca) increased after the addition of the agonist ATP, coupling was unaffected during the period that Ca was maximal. However, coupling decreased transiently approximately 5-10 min after agonist addition when Ca returned to resting levels, indicating that this transient decrease in coupling was unlikely due to a direct action of Ca on gap junctions. An increase in Ca mediated by the ionophore ionomycin that was sustained for several minutes resulted in a more rapid and sustained decrease in coupling (IC(50) ~300 nM Ca(2+), Hill coefficient of 4), indicating that an increase in Ca alone could regulate gap junctions. Thus Ca increases that occurred during agonist stimulation and cell-to-cell Ca(2+) waves were too transient to mediate a sustained uncoupling of lens epithelial cells. PMID- 11502575 TI - Heterologous expression of Na(+)-K(+)-ATPase in insect cells: intracellular distribution of pump subunits. AB - The Na(+)-K(+)-ATPase is a heterodimeric plasma membrane protein responsible for cellular ionic homeostasis in nearly all animal cells. It has been shown that some insect cells (e.g., High Five cells) have no (or extremely low) Na(+)-K(+) ATPase activity. We expressed sheep kidney Na(+)-K(+)-ATPase alpha- and beta subunits individually and together in High Five cells via the baculovirus expression system. We used quantitative slot-blot analyses to determine that the expressed Na(+)-K(+)-ATPase comprises between 0.5% and 2% of the total membrane protein in these cells. Using a five-step sucrose gradient (0.8-2.0 M) to separate the endoplasmic reticulum, Golgi apparatus, and plasma membrane fractions, we observed functional Na(+) pump molecules in each membrane pool and characterized their properties. Nearly all of the expressed protein functions normally, similar to that found in purified dog kidney enzyme preparations. Consequently, the measurements described here were not complicated by an abundance of nonfunctional heterologously expressed enzyme. Specifically, ouabain sensitive ATPase activity, [(3)H]ouabain binding, and cation dependencies were measured for each fraction. The functional properties of the Na(+)-K(+)-ATPase were essentially unaltered after assembly in the endoplasmic reticulum. In addition, we measured ouabain-sensitive (86)Rb(+) uptake in whole cells as a means to specifically evaluate Na(+)-K(+)-ATPase molecules that were properly folded and delivered to the plasma membrane. We could not measure any ouabain sensitive activities when either the alpha-subunit or beta-subunit were expressed individually. Immunostaining of the separate membrane fractions indicates that the alpha-subunit, when expressed alone, is degraded early in the protein maturation pathway (i.e., the endoplasmic reticulum) but that the beta-subunit is processed normally and delivered to the plasma membrane. Thus it appears that only the alpha-subunit has an oligomeric requirement for maturation and trafficking to the plasma membrane. Furthermore, assembly of the alpha-beta heterodimer within the endoplasmic reticulum apparently does not require a Na(+) pump-specific chaperone. PMID- 11502576 TI - Temperature dependence of cloned mammalian and salmonid cardiac Na(+)/Ca(2+) exchanger isoforms. AB - The cardiac Na(+)/Ca(2+) exchanger (NCX), an important regulator of cytosolic Ca(2+) concentration in contraction and relaxation, has been shown in trout heart sarcolemmal vesicles to have high activity at 7 degrees C relative to its mammalian isoform. This unique property is likely due to differences in protein structure. In this study, outward NCX currents (I(NCX)) of the wild-type trout (NCX-TR1.0) and canine (NCX 1.1) exchangers expressed in oocytes were measured to explore the potential contributions of regulatory vs. transport mechanisms to this observation. cRNA was transcribed in vitro from both wild-type cDNA and was injected into Xenopus oocytes. I(NCX) of NCX-TR1.0 and NCX1.1 were measured after 3-4 days over a temperature range of 7-30 degrees C using the giant excised patch technique. The I(NCX) for both isoforms exhibited Na(+)-dependent inactivation and Ca(2+)-dependent positive regulation. The I(NCX) of NCX1.1 exhibited typical mammalian temperature sensitivities with Q(10) values of 2.4 and 2.6 for peak and steady-state currents, respectively. However, the I(NCX) of NCX-TR1.0 was relatively temperature insensitive with Q(10) values of 1.2 and 1.1 for peak and steady-state currents, respectively. I(NCX) current decay was fit with a single exponential, and the resultant rate constant of inactivation (lambda) was determined as a function of temperature. As expected, lambda decreased monotonically with temperature for both isoforms. Although lambda was significantly greater in NCX1.1 compared with NCX-TR1.0 at all temperatures, the effect of temperature on lambda was not different between the two isoforms. These data suggest that the disparities in I(NCX) temperature dependence between these two exchanger isoforms are unlikely due to differences in their inactivation kinetics. In addition, similar differences in temperature dependence were observed in both isoforms after alpha-chymotrypsin treatment that renders the exchanger in a deregulated state. These data suggest that the differences in I(NCX) temperature dependence between the two isoforms are not due to potential disparities in either the I(NCX) regulatory mechanisms or structural differences in the cytoplasmic loop but are likely predicated on differences within the transmembrane segments. PMID- 11502577 TI - Plasma interleukin-6 during strenuous exercise: role of epinephrine. AB - Exercise induces increased levels of plasma interleukin-6 (IL-6) as well as changes in the concentration of lymphocytes and neutrophils. The aim of this study was to investigate a possible role for epinephrine. Seven healthy men participated in an exercise experiment. One month later they received an epinephrine infusion. The exercise consisted of treadmill running at 75% of maximal O(2) consumption for 2.5 h. The infusion trial consisted of 2.5 h of epinephrine infusion calculated to reach the same plasma epinephrine levels seen during the exercise experiment. The plasma concentration of IL-6 increased 29 fold during exercise, with peak levels at the end of exercise. The increase in plasma IL-6 during epinephrine infusion was only sixfold, with the peak value at 1 h after infusion. The lymphocyte concentration increased to the same levels during exercise and epinephrine infusion. The lymphocyte count decreased more in the postexercise period than after epinephrine infusion. The neutrophil concentration was elevated threefold in response to exercise, whereas no change was found in response to epinephrine infusion. In conclusion, the exercise induced increase in plasma IL-6 could not be mimicked by epinephrine infusion. However, epinephrine induced a small increase in IL-6 and may, therefore, partly influence the plasma levels of IL-6 during exercise. In addition, the results support the idea that epinephrine plays a role in exercise-induced changes in lymphocyte number, whereas epinephrine does not mediate exercise-induced neutrocytosis. PMID- 11502578 TI - Decrease in rat taste receptor cell intracellular pH is the proximate stimulus in sour taste transduction. AB - Taste receptor cells (TRCs) respond to acid stimulation, initiating perception of sour taste. Paradoxically, the pH of weak acidic stimuli correlates poorly with the perception of their sourness. A fundamental issue surrounding sour taste reception is the identity of the sour stimulus. We tested the hypothesis that acids induce sour taste perception by penetrating plasma membranes as H(+) ions or as undissociated molecules and decreasing the intracellular pH (pH(i)) of TRCs. Our data suggest that taste nerve responses to weak acids (acetic acid and CO(2)) are independent of stimulus pH but strongly correlate with the intracellular acidification of polarized TRCs. Taste nerve responses to CO(2) were voltage sensitive and were blocked with MK-417, a specific blocker of carbonic anhydrase. Strong acids (HCl) decrease pH(i) in a subset of TRCs that contain a pathway for H(+) entry. Both the apical membrane and the paracellular shunt pathway restrict H(+) entry such that a large decrease in apical pH is translated into a relatively small change in TRC pH(i) within the physiological range. We conclude that a decrease in TRC pH(i) is the proximate stimulus in rat sour taste transduction. PMID- 11502579 TI - Regulation of glutamate transport and transport proteins in a placental cell line. AB - We utilized HRP.1 cells derived from midgestation rat placental labyrinth to determine that the primary pathway for glutamate uptake is via system X, a Na(+) dependent transport system. Kinetic parameters of system X activity were similar to those previously determined in rat and human placental membrane vesicle preparations. Amino acid depletion caused a significant upregulation of system X activity at 6, 24, and 48 h. This increase was reversed by the addition of glutamate and aspartate but not by the addition of alpha-(methylamino)isobutyric acid. Immunoblot analysis of the three transport proteins previously associated with system X activity indicated a trend toward an increase in GLT1, EAAC1, and GLAST1 immunoreactive protein contents by 48 h; cell surface expression of the same was enhanced by 24 h. Inhibition analysis suggested key roles for EAAC1 and GLAST1 in basal anionic amino acid transfer, with an enhanced role for GLT1 under conditions of amino acid depletion. In summary, amino acid availability as well as intracellular metabolism regulate anionic amino acid uptake into this placental cell line. PMID- 11502580 TI - Enhanced mitochondrial testicular antioxidant capacity in Goto-Kakizaki diabetic rats: role of coenzyme Q. AB - Because diabetes mellitus is associated with impairment of testicular function, ultimately leading to reduced fertility, this study was conducted to evaluate the existence of a cause-effect relationship between increased oxidative stress in diabetes and reduced mitochondrial antioxidant capacity. The susceptibility to oxidative stress and antioxidant capacity (in terms of glutathione, coenzyme Q, and vitamin E content) of testis mitochondrial preparations isolated from Goto Kakizaki (GK) non-insulin-dependent diabetic rats and from Wistar control rats, 1 yr of age, was evaluated. It was found that GK mitochondrial preparations showed a lower susceptibility to lipid peroxidation induced by ADP/Fe(2+), as evaluated by oxygen consumption and reactive oxygen species generation. The decreased susceptibility to oxidative stress in diabetic rats was associated with an increase in mitochondrial glutathione and coenzyme Q9 contents, whereas vitamin E was not changed. These results demonstrate a higher antioxidant capacity in diabetic GK rats. We suggest this is an adaptive response of testis mitochondria to the increased oxidative damage in diabetes mellitus. PMID- 11502581 TI - Role of phospholamban in the modulation of arterial Ca(2+) sparks and Ca(2+) activated K(+) channels by cAMP. AB - Phospholamban (PLB) inhibits the sarcoplasmic reticulum (SR) Ca(2+)-ATPase, and this inhibition is relieved by cAMP-dependent protein kinase (PKA)-mediated phosphorylation. The role of PLB in regulating Ca(2+) release through ryanodine sensitive Ca(2+) release channels, measured as Ca(2+) sparks, was examined using smooth muscle cells of cerebral arteries from PLB-deficient ("knockout") mice (PLB-KO). Ca(2+) sparks were monitored optically using the fluorescent Ca(2+) indicator fluo 3 or electrically by measuring transient large-conductance Ca(2+) activated K(+) (BK) channel currents activated by Ca(2+) sparks. Basal Ca(2+) spark and transient BK current frequency were elevated in cerebral artery myocytes of PLB-KO mice. Forskolin, an activator of adenylyl cyclase, increased the frequency of Ca(2+) sparks and transient BK currents in cerebral arteries from control mice. However, forskolin had little effect on the frequency of Ca(2+) sparks and transient BK currents from PLB-KO cerebral arteries. Forskolin or PLB-KO increased SR Ca(2+) load, as measured by caffeine-induced Ca(2+) transients. This study provides the first evidence that PLB is critical for frequency modulation of Ca(2+) sparks and associated BK currents by PKA in smooth muscle. PMID- 11502582 TI - Regulation of PGE(2) and PGI(2) release from human umbilical vein endothelial cells by actin cytoskeleton. AB - Disruption of microfilaments in human umbilical vein endothelial cells (HUVEC) with cytochalasin D (cytD) or latrunculin A (latA) resulted in a 3.3- to 5.7-fold increase in total synthesis of prostaglandin E(2) (PGE(2)) and a 3.4- to 6.5-fold increase in prostacyclin (PGI(2)) compared with control cells. Disruption of the microtubule network with nocodazole or colchicine increased synthesis of PGE(2) 1.7- to 1.9-fold and PGI(2) 1.9- to 2.0-fold compared with control cells. Interestingly, however, increased release of PGE(2) and PGI(2) from HUVEC into the media occurred only when microfilaments were disrupted. CytD treatment resulted in 6.7-fold more PGE(2) and 3.8-fold more PGI(2) released from HUVEC compared with control cells; latA treatment resulted in 17.7-fold more PGE(2) and 11.2-fold more PGI(2) released compared with control cells. Both increased synthesis and release of prostaglandins in response to all drug treatments were completely inhibited by NS-398, a specific inhibitor of cyclooxygenase-2 (COX-2). Disruption of either microfilaments using cytD or latA or of microtubules using nocodazole or colchicine resulted in a significant increase in COX-2 protein levels, suggesting that the increased synthesis of prostaglandins in response to drug treatments may result from increased activity of COX-2. These results, together with studies demonstrating a vasoprotective role for prostaglandins, suggest that the cytoskeleton plays an important role in maintenance of endothelial barrier function by regulating prostaglandin synthesis and release from HUVEC. PMID- 11502583 TI - Regulation of phospholipase C-gamma(1) by the actin-regulatory protein villin. AB - The actin-regulatory protein villin is tyrosine phosphorylated and associates with phospholipase C-gamma(1) (PLC-gamma(1)) in the brush border of intestinal epithelial cells. To study the mechanism of villin-associated PLC-gamma(1) activation, we reconstituted in vitro the tyrosine phosphorylation of villin and its association with PLC-gamma(1). Recombinant villin was phosphorylated in vitro by the nonreceptor tyrosine kinase c-src or by expression in the TKX1 competent cells that carry an inducible tyrosine kinase gene. Using in vitro binding assays, we demonstrated that tyrosine-phosphorylated villin associates with the COOH-terminal Src homology 2 (SH2) domain of PLC-gamma(1). The catalytic activity of PLC-gamma(1) was inhibited by villin in a dose-dependent manner with half maximal inhibition at a concentration of 12.4 microM. Villin inhibited PLC gamma(1) activity by sequestering the substrate phosphatidylinositol 4,5 bisphosphate (PIP(2)), since increasing concentrations of PIP(2) reversed the inhibitory effects of villin on PLC activity. The inhibition of PLC-gamma(1) activity by villin was reversed by the tyrosine phosphorylation of villin. Further, we demonstrated that tyrosine phosphorylation of villin abolished villin's ability to associate with PIP(2). In conclusion, tyrosine-phosphorylated villin associates with the COOH-terminal SH2 domain of PLC-gamma(1) and activates PLC-gamma(1) catalytic activity. Villin regulates PLC-gamma(1) activity by modifying its own ability to bind PIP(2). This study provides biochemical proof of the functional relevance of tyrosine phosphorylation of villin and identifies the molecular mechanisms involved in the activation of PLC-gamma(1) by villin. PMID- 11502584 TI - Protein kinase Cepsilon contributes to regulation of the sarcolemmal Na(+)-K(+) pump. AB - A reduction in angiotensin II (ANG II) in vivo by treatment of rabbits with the angiotensin-converting enzyme inhibitor, captopril, increases Na(+)-K(+) pump current (I(p)) of cardiac myocytes. This increase is abolished by exposure of myocytes to ANG II in vitro. Because ANG II induces translocation of the epsilon isoform of protein kinase C (PKCepsilon), we examined whether this isozyme regulates the pump. We treated rabbits with captopril, isolated myocytes, and measured I(p) of myocytes voltage clamped with wide-tipped patch pipettes. I(p) of myocytes from captopril-treated rabbits was larger than I(p) of myocytes from controls. ANG II superfusion of myocytes from captopril-treated rabbits decreased I(p) to levels similar to controls. Inclusion of PKCepsilon-specific blocking peptide in pipette solutions used to perfuse the intracellular compartment abolished the effect of ANG II. Inclusion of psiepsilonRACK, a PKCepsilon specific activating peptide, in pipette solutions had an effect on I(p) that was similar to that of ANG II. There was no additive effect of ANG II and psiepsilonRACK. We conclude that PKCepsilon regulates the sarcolemmal Na(+)-K(+) pump. PMID- 11502585 TI - Peroxynitrite causes endothelial cell monolayer barrier dysfunction. AB - Nitric oxide (.NO) attenuates hydrogen peroxide (H(2)O(2))-mediated barrier dysfunction in cultured porcine pulmonary artery endothelial cells (PAEC) (Gupta MP, Ober MD, Patterson C, Al-Hassani M, Natarajan V, and Hart, CM. Am J Physiol Lung Cell Mol Physiol 280: L116-L126, 2001). However,.NO rapidly combines with superoxide (O) to form the powerful oxidant peroxynitrite (ONOO(-)), which we hypothesized would cause PAEC monolayer barrier dysfunction. To test this hypothesis, we treated PAEC with ONOO(-) (500 microM) or 3-morpholinosydnonimine hydrochloride (SIN-1; 1-500 microM). SIN-1-mediated ONOO(-) formation was confirmed by monitoring the oxidation of dihydrorhodamine 123 to rhodamine. Both ONOO(-) and SIN-1 increased albumin clearance (P < 0.05) in the absence of cytotoxicity and altered the architecture of the cytoskeletal proteins actin and beta-catenin as detected by immunofluorescent confocal imaging. ONOO(-)-induced barrier dysfunction was partially reversible and was attenuated by cysteine. Both ONOO(-) and SIN-1 nitrated tyrosine residues, including those on beta-catenin and actin, and oxidized proteins in PAEC. The introduction of actin treated with ONOO(-) into PAEC monolayers via liposomes also resulted in barrier dysfunction. These results indicate that ONOO(-) directly alters endothelial cytoskeletal proteins, leading to barrier dysfunction. PMID- 11502586 TI - Mechanism of increased renal gene expression during metabolic acidosis. AB - Increased renal catabolism of plasma glutamine during metabolic acidosis generates two ammonium ions that are predominantly excreted in the urine. They function as expendable cations that facilitate the excretion of acids. Further catabolism of alpha-ketoglutarate yields two bicarbonate ions that are transported into the venous blood to partially compensate for the acidosis. In rat kidney, this adaptation is sustained, in part, by the induction of multiple enzymes and various transport systems. The pH-responsive increases in glutaminase (GA) and phosphoenolpyruvate carboxykinase (PEPCK) mRNAs are reproduced in LLC PK(1)-fructose 1,6-bisphosphatase (FBPase) cells. The increase in GA activity results from stabilization of the GA mRNA. The 3'-untranslated region of the GA mRNA contains a direct repeat of an eight-base AU sequence that functions as a pH response element. This sequence binds zeta-crystallin/NADPH:quinone reductase with high affinity and specificity. Increased binding of this protein during acidosis may initiate the pH-responsive stabilization of the GA mRNA. In contrast, induction of PEPCK occurs at the transcriptional level. In LLC-PK(1) FBPase(+) kidney cells, a decrease in intracellular pH leads to activation of the p38 stress-activated protein kinase and subsequent phosphorylation of transcription factor ATF-2. This transcription factor binds to cAMP-response element 1 within the PEPCK promoter and may enhance its transcription during metabolic acidosis. PMID- 11502587 TI - Trafficking and cell surface stability of ENaC. AB - The epithelial Na(+) channel (ENaC) plays a key role in the regulation of Na(+) and water absorption in several epithelia, including those of the distal nephron, distal colon, and lung. Accordingly, mutations in ENaC leading to reduced or increased channel activity cause human diseases such as pseudohypoaldosteronism type I or Liddle's syndrome, respectively. The gain of ENaC function in Liddle's syndrome is associated with increased activity and stability of the channel at the plasma membrane. Thus understanding the regulation of channel processing and trafficking to and stability at the cell surface is of fundamental importance. This review describes some of the recent advances in our understanding of ENaC trafficking, including the role of glycosylation, ENaC solubility in nonionic detergent, targeting signal(s) and hormones. It also describes the regulation of ENaC stability at the cell surface and the roles of the ubiquitin ligase Nedd4 (and ubiquitination) and clathrin-mediated endocytosis in that regulation. PMID- 11502588 TI - Cloning and characterization of the human urea transporter UT-A1 and mapping of the human Slc14a2 gene. AB - We have isolated and characterized the human homolog of the rat largest urea transporter of the UT-A family (hUT-A1). The 4.2-kb hUT-A1 cDNA encodes a 920 amino acid peptide, which is 89% identical to the rat UT-A1 protein. By Northern hybridization, hUT-A1 expression is detected in the human inner medulla as a approximately 4.4-kb mRNA transcript. By Western analysis, hUT-A1 is identified as a approximately 100-kDa protein in the human inner medulla. By immunohistochemistry, hUT-A1 expression is localized to the inner medullary collecting duct (IMCD). When transfected into HEK-293 cells hUT-A1 cDNA is translated into a approximately 98-kDa protein. Expression of hUT-A1 in Xenopus oocytes results in phloretin-inhibitable uptake of (14)C-urea, which shows only modest stimulation by cAMP, suggesting that in the human IMCD vasopressin may have a limited role in the short-term regulation of hUT-A1-mediated urea transport. We determined the organization of the human Slc14a2 gene and identified 20 exons distributed over approximately 67.5 kb on chromosome 18, from which hUT-A1 and the other human urea transporter, hUT-A2, are transcribed. PMID- 11502589 TI - Odd E. Hanssen and the Hanssen method for measurement of single-nephron glomerular filtration rate. AB - In the middle of the twentieth century, the suspicion that deep and superficial nephrons might serve different functions created a demand for measurement of single-nephron glomerular filtration rate (SNGFR). Rather unexpectedly, the answer came from Odd E. Hanssen (1917-1964), a Norwegian physician working on his own in the Department of Pathological Anatomy, University of Oslo, with minimal support and no interaction with renal physiologists. In 1963, after nearly 10 years of work, he presented the ferrocyanide method, allowing simultaneous estimates of SNGFR in a large number of nephrons in all layers of the kidney. This review first describes his early visions of the method and the elaborate and extremely time-consuming studies in mice to verify the technique. As a byproduct came valuable information on the relationship between nephron size and SNGFR, glomerular intermittency, and the emptying of the tubules on filtration stop. Hanssen died from a cerebral hemorrhage in 1964, and for several years the method seemed entirely forgotten. Fortunately, Andrew Baines took up the use of ferrocyanide in 1963-1964 while working on his thesis in Toronto, but his first publication came in 1969 from Saclay, France, in collaboration with Christian de Rouffignac. Modifications allowing determination of absolute SNGFR were worked out by de Rouffignac and by Jaime Coehlo in New York. Thereafter, the "Hanssen method" spread rapidly, and in the early 1980s about 50 reports had been published from 17 laboratories in 9 countries. The distribution of SNGFR in mammals, birds, and fish was described, as well as the response to water and salt loads, vasoactive substances, hormones, varying perfusion pressure, blood loss, etc. Finally, after mentioning two recent methods inspired by the Hanssen technique but using other filtration markers, the review concludes that most of our present knowledge on SNGFR distribution and regulation has been obtained by the method developed by Hanssen 40 years ago. PMID- 11502590 TI - Mechanism underlying diuretic effect of L-NAME at a subpressor dose. AB - The diuretic effects of nitric oxide (NO) synthase inhibitors administered at subpressor dose in rats are controversial, and the tubular segments involved are not known. In the present study, we examined the effect of N(omega)-nitro-L arginine methyl ester (L-NAME) at a subpressor dose on renal interstitial NO and cGMP activity and on renal tubular segmental reabsorption of fluid in the rat. Intravenous infusion of L-NAME at 1 microg. kg(-1). min(-1) in Sprague-Dawley rats (N = 8), which did not alter mean arterial pressure or glomerular filtration rate, significantly increased urine flow rate (U(v); from 78.2 +/- 12.7 to 117.1 +/- 14.9 microl/min, P < 0.05). Paradoxically, this effect of L-NAME was concomitant with significant increases in nitrite/nitrate (from 10.79 +/- 1.20 to 16.50 +/- 2.60 microM, P < 0.05) and cGMP (from 0.65 +/- 0.09 to 0.98 +/- 0.18 nM, P < 0.05) concentrations in renal cortical microdialysate as well as the nitrite/nitrate concentration in the medullary microdialysate. Micropuncture studies in the superficial nephron revealed that L-NAME significantly increased the flow rate (from 8.3 +/- 0.9 to 12.2 +/- 1.2 nl/min, P < 0.05) and fractional delivery of fluid to the distal tubule, but not those in the late proximal tubule. In conclusion, L-NAME, at the subpressor dose used in this study, increased renal nitrate/nitrite and cGMP and inhibited fluid reabsorption in tubular segments between the late proximal tubule and the distal tubule of superficial nephrons. PMID- 11502591 TI - Role of guanylyl cyclase and cytochrome P-450 on renal response to nitric oxide. AB - The present study evaluated whether inhibition of guanylyl cyclase (GC) with 1H (1,2,4)oxadiazolo[4,3-a]quinoxaline-1-one (ODQ) and methylene blue (MB) or inhibition of the renal metabolism of arachidonic acid by cytochrome P-450 (CYP450) enzymes with 1-aminobenzotriazole (ABT) and N-hydroxy-N'-(4 butyl-2 methyl phenyl)formamidine (HET0016) alters the renal tubular and vascular effects of a nitric oxide (NO) donor in vivo. Intrarenal infusion of ODQ or MB at a dose of 170 nmol. kg(-1). min(-1) lowered renal blood flow (RBF) by 30 and 15%, respectively; glomerular filtration rate (GFR) by 26 and 18%, respectively; and sodium and water excretion by approximately 35%. In rats pretreated with nitro-L arginine methyl ester (37 nmol. kg(-1). min(-1)) to block the endogenous production of NO, intrarenal infusion of the NO donor S-nitroso-N-acetylcysteine (S-NO-NAC; 50 nmol. kg(-1). min(-1)) increased RBF (18%), sodium (73%), and water excretion (61%). ODQ or MB administration blocked the effect of S-NO-NAC on RBF but not the diuretic and natriuretic response. Pretreatment of rats with ABT or HET0016 also abolished the renal vasodilatory response to the NO donor and reduced its diuretic and natriuretic effect. These results indicate that both activation of GC and inhibition of CYP450 enzymes contribute to the renal vascular actions of NO, whereas the natriuretic and diuretic actions of NO appear to be largely CYP450 dependent. PMID- 11502592 TI - Vitamin D reduces renal NaPi-2 in PTH-infused rats: complexity of vitamin D action on renal P(i) handling. AB - Acute administration of dihydroxycholecalciferol [1,25(OH)(2)D(3)] blunts phosphaturia and increases urinary cAMP excretion in parathyroid hormone (PTH) infused parathyroidectomized (PTX) rats. Because chronic administration of 1,25(OH)(2)D(3) enhances the phosphaturic response to exogenous parathyroid hormone despite blunting of urinary cAMP excretion, we have examined the expression of the renal cortex type II Na-P(i) cotransporter (NaPi-2) mRNA and protein in 1) chronic PTX Sabra rats, 2) PTX rats receiving a physiological dose of 1,25(OH)-2-D(3), 3) PTX rats receiving 35 ng/h of PTH, and 4) rats receiving both PTH and 1,25(OH)(2)D(3), for 7 days via osmotic minipumps. Our results confirm that there is increased phosphaturia in the PTH+1,25(OH)(2)D(3)-infused animals despite blunting of urinary cAMP excretion, a reduced filtered load of phosphate, and lack of a phosphaturic effect by 1,25(OH)(2)D(3) alone. Both PTH and 1,25(OH)(2)D(3) significantly reduced expression of renal cortex NaPi-2 mRNA and NaPi-2 protein, and the administration of PTH together with 1,25(OH)(2)D(3) had additive effects in further decreasing NaPi-2 mRNA and NaPi-2 protein levels. Expression of two other epithelial transporters, type 1 Na-sulfate and type 1 Na glucose cotransporters, were not different between the groups, suggesting specificity of the effects of PTH and 1,25(OH)(2)D(3) on phosphate transport. The effect of chronic administration of 1,25(OH)(2)D(3) has not been noted previously, and the cellular mechanisms and signaling processes that mediate the decrease in NaPi-2 remain to be determined. PMID- 11502593 TI - CFTR disruption impairs cAMP-dependent Cl(-) secretion in primary cultures of mouse cortical collecting ducts. AB - The role of the cystic fibrosis transmembrane conductance regulator (CFTR) in the renal cortical collecting duct (CCD) has not yet been fully elucidated. Here, we investigated the effects of deamino-8-D-arginine vasopressin (dDAVP) and isoproterenol (ISO) on NaCl transport in primary cultured CCDs microdissected from normal [CFTR(+/+)] and CFTR-knockout [CFTR(-/-)] mice. dDAVP stimulated the benzamyl amiloride (BAm)-sensitive transport of Na(+) assessed by the short circuit current (I(sc)) method in both CFTR(+/+) and CFTR(-/-) CCDs to a very similar degree. Apical addition of 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) or glibenclamide partially inhibited the rise in I(sc) induced by dDAVP and ISO in BAm-treated CFTR(+/+) CCDs, whereas dDAVP, ISO, and NPPB did not alter I(sc) in BAm-treated CFTR(-/-) CCDs. dDAVP stimulated the apical-to-basal flux and, to a lesser extent, the basal-to-apical flux of (36)Cl(-) in CFTR(+/+) CCDs. dDAVP also increased the apical-to-basal (36)Cl(-) flux in CFTR(-/-) CCDs but not the basal-to-apical (36)Cl(-) flux. These results demonstrate that CFTR mediates the cAMP-stimulated component of secreted Cl(-) in mouse CCD. PMID- 11502594 TI - Collagen IV promotes repair of renal cell physiological functions after toxicant injury. AB - Collagen IV is found in the renal proximal tubular cell (RPTC) basement membrane and is a mediator of renal development and function. Pharmacological concentrations of L-ascorbic acid phosphate (AscP) promote the repair of physiological functions in RPTC sublethally injured by S-(1,2-dichlorovinyl)-L cysteine (DCVC). We hypothesized that AscP promotes RPTC repair by stimulating collagen IV synthesis and/or deposition. RPTC exhibited increased synthesis but decreased deposition of collagen IV after DCVC exposure. In contrast, RPTC cultured in pharmacological concentrations of AscP maintained collagen IV deposition. The activity of prolyl hydroxylase was decreased in RPTC after DCVC injury, an effect that was partially attenuated in injured RPTC cultured in pharmacological concentrations of AscP. The addition of exogenous collagen IV to the culture media of DCVC-injured RPTC promoted the repair of mitochondrial function and Na(+)-K(+)-ATPase activity. However, neither collagen I, laminin, nor fibronectin promoted cell repair. These data demonstrate an association between AscP-stimulated deposition of collagen IV and exogenous collagen IV and repair of physiological functions, suggesting that collagen IV plays a specific role in RPTC repair after sublethal injury. PMID- 11502595 TI - Interaction of cations, anions, and weak base quinine with rat renal cation transporter rOCT2 compared with rOCT1. AB - The rat organic cation transporter (rOCT)-2 was characterized by electrical and tracer flux measurements compared with rOCT1. By applying choline gradients to voltage-clamped Xenopus oocytes expressing rOCT2, potential-dependent currents could be induced in both directions. Tracer flux measurements with seven organic cations revealed similar Michaelis-Menten constant values for both transporters, with the exception of guanidine. In parallel experiments with rOCT2 and rOCT1, inhibition of tetraethylammonium transport by 12 cations, 2 weak bases, corticosterone, and the anions para-amminohippurate, alpha-ketoglutarate, and probenecid was characterized. The IC(50) values of many inhibitors were similar for both transporters, whereas others were significantly different. Mepiperphenidol and O-methylisoprenaline showed an approximately 70-fold lower and corticosterone a 38-fold higher affinity for rOCT2. With the use of these inhibitors together with previous information on cation transporters, experimental protocols are proposed to dissect out the individual contributions of rOCT2 and rOCT1 in intact proximal tubule preparations. Inhibition experiments at different pH levels strongly suggest that the weak base quinine passively permeates the plasma membrane at physiological pH and inhibits rOCT2 from the intracellular side. PMID- 11502596 TI - Distinct characteristics of two human Nedd4 proteins with respect to epithelial Na(+) channel regulation. AB - The epithelial Na(+) channel (ENaC) is regulated via PY motif-WW domain interaction by the mouse (m) ubiquitin-protein ligase mNedd4-2 but not by its close relative mNedd4-1. Whereas mNedd4-1 is composed of one C2, three WW, and one HECT domain, mNedd4-2 comprises four WW domains and one HECT domain. Both proteins have human (h) homologs, hNedd4-1 and hNedd4-2; however, both of them include four WW domains. Therefore, we characterized hNedd4-1 and hNedd4-2 in Xenopus laevis oocytes with respect to ENaC binding and interaction. We found that hNedd4-2 binds to and abrogates ENaC activity, whereas hNedd4-1 does not coimmunoprecipitate with ENaC and has only modest effects on ENaC activity. Structure-function studies revealed that the C2 domain of hNedd4-1 prevents this protein from downregulating ENaC and that WW domains 3 and 4, involved in interaction with ENaC, do not by themselves provide specificity for ENaC recognition. Taken together, our data demonstrate that hNedd4-2 inhibits ENaC, implying that this protein is a modulator of salt homeostasis, whereas hNedd4-1 is not primarily involved in ENaC regulation. PMID- 11502597 TI - Transport of plasma proteins across vasa recta in the renal medulla. AB - In this study, we have extended a mathematical model of microvascular exchange in the renal medulla to elucidate the mechanisms by which plasma proteins are transported between vasa recta and the interstitium. In contrast with other work, a distinction was made between the paracellular pathway and the transcellular route (i.e., water channels) in descending vasa recta (DVR). Our model first indicates that concentration polarization on the interstitial side of vasa recta has a negligible effect on medullary function. Our results also suggest that, whereas proteins are cleared from the interstitium by convection, both diffusion and convection play a role in carrying proteins to the interstitium. In those regions where transcapillary oncotic pressure gradients favor volume influx through the paracellular pathway in DVR, diffusion is the only means by which proteins can penetrate the interstitium. Whether the source of interstitial protein is DVR or ascending vasa recta depends on medullary depth, vasa recta permeability to proteins, and vasa recta reflection coefficients to small solutes and proteins. Finally, our model predicts significant axial protein gradients in the renal medullary interstitium. PMID- 11502598 TI - Ammonium interaction with the epithelial sodium channel. AB - The purpose of this study was to investigate the direct effect of NH(3)/NH on mouse epithelial Na(+) channels (mENaC) expressed in Xenopus oocytes. Two electrode voltage-clamp and ion-selective microelectrodes were used to measure the Na(+) current, intracellular pH (pH(i)), and ion activities in oocytes expressing mENaC. In oocytes expressing mENaC, removal of external Na(+) reversibly hyperpolarized membrane potential by 129 +/- 5.3 mV in the absence of 20 mM NH(4)Cl but only by 100 +/- 7.8 mV in its presence. Amiloride completely inhibited the changes in membrane potential. In oocytes expressing mENaC, butyrate (20 mM) caused a decrease in pH(i) (0.43 +/- 0.07) similar to the NH(4)Cl-induced pH(i) decrease (0.47 +/- 0.12). Removal of Na(+) in the presence of butyrate caused hyperpolarization that was not significantly different from that in the absence of butyrate at high pH(i) (in the absence of NH(4)Cl). Removal of external Na(+) resulted in an outward current of 3.7 +/- 0.8 microA (at -60 mV). The magnitude of this change in current was only 2.7 +/- 0.7 microA when Na(+) was removed in the presence of NH(4)Cl. In oocytes expressing mENaC, NH(4)Cl also caused a decrease in whole cell conductance at negative potential and an outward current at positive potential. In the presence of amiloride, steady-state current and the change in current caused by removal of Na(+) were not different from zero. These results indicate that NH(4)Cl inhibits Na(+) transport when mENaC is expressed in oocytes. The inhibition of voltage changes is not due to intracellular acidification caused by NH(4)Cl. Permeability and selectivity of ENaC to NH may play a role. PMID- 11502599 TI - Puromycin aminonucleoside damages the glomerular size barrier with minimal effects on charge density. AB - Puromycin aminonucleoside (PAN) has been suggested to reduce glomerular charge density, to create large glomerular "leaks," or not to affect the glomerular barrier. Therefore, we analyzed glomerular charge and size selectivity in vivo and in isolated kidneys perfused at 8 degrees C (cIPK) in control and PAN-treated rats. The fractional clearances (theta) for albumin and Ficoll of similar hydrodynamic size were 0.0017 +/- 0.0004 and 0.15 +/- 0.02, respectively, in control cIPKs. Two-pore analysis gave similar results in vivo and in vitro, with small- and large-pore radii of 47-52 and 85-105 A, respectively, in controls. Puromycin increased the number of large pores 40-50 times, the total pore area over diffusion distance decreased by a factor of 25-30, and the small-pore radius increased by 33% (P < 0.001 for all comparisons of size selectivity and theta). The effect of PAN was less dramatic on the estimated wall charge density, which was 73% of that of controls. We conclude that puromycin effectively destroys the glomerular size barrier with minimal effects on charge density. PMID- 11502600 TI - Quantification of TGF-beta1 mRNA along rat nephron in obstructive nephropathy. AB - Unilateral ureteral obstruction (UUO) leads to interstitial fibrosis of the obstructed kidney, and transforming growth factor-beta1 (TGF-beta1) is thought to play an important role in this process. Although increased TGF-beta1 mRNA expression in the obstructed kidney has been demonstrated, the source of the increased TGF-beta1 remains to be elucidated. To determine the precise localization of TGF-beta1 in the obstructed kidney, we examined TGF-beta1 mRNA expression using in situ hybridization and competitive RT-PCR in rats with UUO. In situ hybridization demonstrated that TGF-beta1 mRNA expression was preferentially increased in tubular epithelial cells and to a lesser degree in infiltrating macrophages in obstructed kidneys. Quantitative analysis using competitive RT-PCR in microdissected nephron segments revealed that levels of TGF beta1 mRNA in obstructed kidneys relative to control kidneys increased significantly in proximal tubules, thick ascending limbs of Henle, and distal convoluted tubules, whereas those in glomeruli and collecting ducts did not change significantly. Of the tubular segments, the proximal tubules appeared to predominantly contribute to increased TGF-beta1 mRNA. Our findings suggest that renal tubules, particularly proximal tubules, are the main contributors to increased TGF-beta1 mRNA expression in obstructed kidneys and to the subsequent interstitial fibrosis. PMID- 11502601 TI - p53 Protects renal inner medullary cells from hypertonic stress by restricting DNA replication. AB - We previously found that p53 upregulation by hypertonicity protected renal inner medullary collecting duct (mIMCD3) cells from apoptosis. The purpose of the present study was to investigate the mechanism by which p53 protects the cells. We now find that hypertonicity (NaCl added to a total of 500 mosmol) inhibits DNA replication and delays G(1)-S transition as concluded from analysis of cell cycle distributions and bromodeoxyuridine (BrDU) incorporation rates. Lowering of p53 with p53 antisense oligonucleotide attenuated such effects of hypertonicity, resulting in an increased number of apoptotic cells in S phase and cells with >4 N DNA. Results with synchronized cells are similar, showing that cells in the early S phase are more sensitive to hypertonicity. Immunocytochemistry revealed that p53 becomes phosphorylated on Ser(15) and translocates to the nucleus in S both in isotonic and hypertonic conditions. Caffeine (2 mM) greatly reduces the p53 level and Ser(15) phosphorylation, followed by a remarkable increase of DNA replication rate, by failure of hypertonicity to inhibit it, and by reduction of cell number during hypertonicity. Finally, inhibition of DNA replication by the DNA polymerase inhibitor aphidicolin significantly improves cell survival, confirming that keeping cells in G(1) and decreasing the rate of DNA replication is protective and that these actions of p53 most likely are what normally help protect cells against hypertonicity. PMID- 11502602 TI - Immunocytochemical localization of Na-K-ATPase alpha- and gamma-subunits in rat kidney. AB - The gamma-subunit of the Na-K-ATPase is a single-span membrane protein that alters the kinetic properties of the enzyme. It is expressed in the kidney, but our initial observations indicated that it is not present in all nephron segments (Arystarkhova E, Wetzel RK, Asinovski NK, and Sweadner KJ. J Biol Chem 274: 33183 33185, 1999). Here we used triple-label confocal immunofluorescence microscopy in rat kidney with antibodies to Na-K-ATPase alpha1- and gamma-subunits and nephron segment-specific markers. Na-K-ATPase alpha1-subunit stain was low but unambiguous in proximal segments, moderate in macula densa, connecting tubules, and cortical collecting ducts, high in thick ascending limb and distal convoluted tubules, and nearly undetectable in glomeruli, descending and ascending thin limb, and medullary collecting ducts. The gamma-subunit colocalized at staining levels similar to alpha1-subunit in basolateral membranes in all segments except cortical thick ascending limb and cortical collecting ducts, which had alpha1 subunit but no detectable gamma-subunit stain. Selective gamma-subunit expression may contribute to the variations in Na-K-ATPase properties in different renal segments. PMID- 11502603 TI - SNAP-25-associated Hrs-2 protein colocalizes with AQP2 in rat kidney collecting duct principal cells. AB - The vasopressin-induced trafficking of aquaporin-2 (AQP2) water channels in kidney collecting duct is likely mediated by vesicle-targeting proteins (N ethylmaleimide-sensitive factor attachment protein receptors). Hrs-2 is an ATPase believed to have a modulatory role in regulated exocytosis. To examine whether Hrs-2 is expressed in rat kidney, we carried out RT-PCR combined with DNA sequence analysis and Northern blotting using a digoxigenin-labeled Hrs-2 RNA probe. RT-PCR and Northern blotting revealed that Hrs-2 mRNA is localized in all zones of rat kidney. The presence of Hrs-2 protein in rat kidney was confirmed by immunoblotting, revealing a 115-kDa protein in kidney and brain membrane fractions corresponding to the expected molecular size of Hrs-2. Immunostaining and confocal laser scanning microscopy of LLC-PK(1) cells (a porcine proximal tubule cell line) transfected with Hrs-2 DNA confirmed the specificity of the antibody and revealed that Hrs-2 is mainly localized in intracellular compartments, including cathepsin D-containing lysosomal/endosomal compartments. The cellular and subcellular localization of Hrs-2 in rat kidney was examined by immunocytochemistry and confocal laser scanning microscopy. Hrs-2 immunoreactivity was observed in collecting duct principal cells, and weaker labeling was detected in other nephron segments. The labeling was predominantly present in intracellular vesicles, but labeling was also observed in the apical plasma membrane domains of some cells. Colabeling with AQP2 revealed colocalization in vesicles and apical plasma membrane domains, suggesting a role for Hrs-2 in regulated AQP2 trafficking. PMID- 11502604 TI - Neurotransmitter-stimulated ion transport by cultured porcine vas deferens epithelium. AB - A collagenase-based dissociation technique has been developed to routinely establish monolayer cultures of freshly isolated porcine vas deferens epithelium. Cells isolated from each tissue are transferred to 25-cm(2) tissue culture flasks and grown in a standard cell culture medium. Flasks reach confluency in 3-4 days, and cells are subsequently seeded onto permeable supports. Cultured cells display a monolayer cobblestone appearance and are immunoreactive to anti-ZO-1 and anti cytokeratin antibodies. Electron microscopy is employed to demonstrate the presence of junctional complexes and microvilli. When evaluated in modified Ussing chambers, cultured monolayers exhibit a basal lumen negative potential difference, high electrical resistance (>1,000 Omega. cm(2)), and respond to norepinephrine, vasopressin, ATP, adenosine, and histamine, with changes in short circuit current indicative of anion secretion. Responses are significantly attenuated in Cl(-)- and/or HCO-free solutions. Attempts to further optimize culture conditions have shown that chronic exposure to insulin increases proliferation rates. Thus the culture method described will reliably produce viable neurotransmitter-responsive cell monolayers that will allow for the characterization of vas deferens epithelial function and associated control mechanisms. PMID- 11502605 TI - In vivo visualization of characteristics of renal microcirculation in hypertensive and diabetic rats. AB - We developed a videomicroscope system with a charge-coupled device camera and evaluated it in the investigation of the glomerular microcirculation in normal [Wistar-Kyoto (WKY)], spontaneously hypertensive (SHR), and streptoyotocin induced diabetic rats (STZ). In WKY, the diameter of the afferent arterioles (Af) was 11.9 +/- 0.7 microm and that of the efferent arterioles (Ef) was 8.9 +/- 0.7 microm. Af and Ef in each glomerulus could be visualized simultaneously with continuous recording of blood pressure and renal blood flow. In SHR, Af diameter was constricted to approximately 60% of that in WKY. A dose-dependent dilation of Af and Ef was observed after administration of barnidipine (1-10 microg/kg iv), a calcium channel antagonist, in all three groups. No change was seen in the Af-to Ef diameter ratio (Af/Ef ratio) in WKY. In SHR, the Af/Ef ratio increased significantly because of the marked dilation of Af after barnidipine administration. In contrast, barnidipine dilated Ef in STZ, causing a significant reduction in the Af/Ef ratio. This system can analyze in vivo glomerular microcirculation and systemic macrocirculation simultaneously, allowing more direct investigation of the characteristics of and acute changes in glomerular microcirculation in pathological animals. PMID- 11502607 TI - Chronic recurrent multifocal osteomyelitis is a differential diagnosis of juvenile idiopathic arthritis. PMID- 11502606 TI - Radiographic scoring methods as outcome measures in rheumatoid arthritis: properties and advantages. AB - BACKGROUND: Use of scored radiographs as an outcome measure can help estimate the progression of rheumatoid arthritis (RA). Radiographs not only provide permanent records with which to evaluate RA serially, but can also be randomised and blinded, a major advantage in clinical trials. OBJECTIVES AND METHOD: Medline was searched for information about the principal methods of assessing joints affected by RA. Each technique was evaluated for its measurement properties, advantages, and limitations. MAIN FINDINGS: The most commonly used methods are those devised by Sharp, Larsen, and van der Heijde/Sharp, and their variants. Methods based on the Sharp technique provide separate scores for erosion and for joint space narrowing. Larsen and variants, together with the Simple Erosion Narrowing Score (SENS) method, provide an overall score. Each method's measurement properties (feasibility, time consumption, etc) depend on the degree of detail it considers. Authors consistently recommend taking a posteroanterior view of hand and foot radiographs, and the use of trained raters. Intra- and interrater reliability values are generally higher than 0.70 (less often assessed by the intraclass correlation coefficient than the correlation coefficient). Sensitivity to change is calculated by several techniques (standardised response mean (SRM), adjusted SRM, minimal detectable change, smallest detectable difference). Most methods assessed with SRM reach a value of 0.80 or more. CONCLUSION: Standardised procedures are available for performing and reading radiographs in RA. The choice of scoring method depends on the time and staff available, and the required degree of reliability and sensitivity to change. PMID- 11502608 TI - Standing committee on epidemiology and health services research. PMID- 11502609 TI - Responsiveness of the WOMAC osteoarthritis index as compared with the SF-36 in patients with osteoarthritis of the legs undergoing a comprehensive rehabilitation intervention. AB - OBJECTIVE: To compare the responsiveness of the condition-specific Western Ontario and McMaster Universities osteoarthritis (OA) index (WOMAC) and the generic Short Form-36 (SF-36) in patients with OA of the legs undergoing a comprehensive inpatient rehabilitation intervention. METHODS: A prospective follow up study of consecutively referred inpatients of a rehabilitation clinic was made. The patients included fulfilled the American College of Rheumatology criteria for knee or hip OA and underwent both passive and, particularly, active physical therapy for three to four weeks. Responsiveness assessment was performed using the standardised response mean (SRM), effect size, and Guyatt's responsiveness statistic between admission and discharge (end of rehabilitation) and then again between admission and three months later. For pain and function the SRMs were stratified by sex and OA joint. Effects were tested by the t test and SRMs of different scales were compared by the jack knife test. RESULTS: At the three month follow up, complete data were obtained for 223 patients. In general, the three responsiveness statistics showed a similar order of responsiveness. For both instruments, the pain scales were more responsive than the function scales. The responsiveness of the pain scale of both instruments was comparable (SRM=0.723 for WOMAC and SRM=0.528 for SF-36 at the end of rehabilitation; SRM=0.377 for WOMAC and SRM=0.468 for SF-36 at the three month follow up). In the measurement of function, the WOMAC was significantly more responsive than the SF-36 (SRMs, end of rehabilitation: 0.628 v 0.249; three month follow up: 0.235 v -0.001). Responsiveness tended to be higher in women and in knee OA than in men and hip OA. CONCLUSIONS: Both instruments, the WOMAC and the SF-36, capture improvement in pain in patients undergoing comprehensive inpatient rehabilitation intervention. Functional improvement can be detected better by the WOMAC than by the SF-36. All the other scales of both instruments were more weakly responsive. PMID- 11502610 TI - Health Assessment Questionnaire modifications: is standardisation needed? AB - BACKGROUND: Physical disability is part of the end point measures in rheumatoid arthritis clinical trials. The Stanford Health Assessment Questionnaire Disability Index (HAQ DI) is often used for this purpose but lacks international uniformity owing to variations in the translated and adapted questionnaires and variations in its calculation. To study the consequences of these variations the previous Dutch HAQ (HAQ90) was revised, resulting in a new Dutch HAQ (HAQ99). OBJECTIVE: To compare DI scores from the two versions, and to study the consequences of applying different calculation methods for the DI score. METHODS: 78 patients completed both the HAQ99 and the HAQ90. To compare the use of different category score calculation methods a post hoc analysis on prospectively collected data obtained in clinical trials was performed. RESULTS: No statistically significant differences were observed between the DI scores of the HAQ90 and the HAQ99 using the alternative method (that is, without correcting for aid and devices). However, correcting for the use of aid or devices or not did result in statistically significant different DI scores. The systematic shift when using the maximum or mean item score for calculation of the category score resulted in non-comparable absolute DI scores. CONCLUSION: The use of HAQ DI questionnaires with different numbers of items and/or categories does not hinder international comparability, except when these variations interfere with the calculation method of the DI (as in the case of questionnaires without a section correcting for devices). For the sake of international uniformity the HAQ or any validated translation should be used and calculated in a standard way, including correcting for the use of aid and devices, and taking the maximum within each category as the category score. PMID- 11502611 TI - Serum TIMP-1, TIMP-2, and MMP-1 in patients with systemic sclerosis, primary Raynaud's phenomenon, and in normal controls. AB - BACKGROUND: Excess tissue matrix accumulates in systemic sclerosis (SSc), accounting for both visceral and dermal fibrosis. It is suggested that decreased serum levels of matrix metalloproteinases (MMPs) or increased levels of tissue inhibitors of matrix metalloproteinases (TIMPs) may account for this matrix accumulation. OBJECTIVE: To measure serum levels of tissue inhibitors of metalloproteinases, TIMP-1, TIMP-2, and collagenase-1 (MMP-1), in patients with diffuse cutaneous systemic sclerosis (dcSSc), limited cutaneous systemic sclerosis (lcSSc), primary Raynaud's phenomenon (RP), and in normal controls. METHODS: Serum samples from patients with dcSSc (n=83), lcSSc (n=87), RP (n=80), and normal controls (n=98) were analysed using enzyme linked immunosorbent assays (ELISAs) for total TIMP-1, TIMP-2, and MMP-1. Results from each assay were analysed by the Kruskal-Wallis test. Dunn's multiple comparison post-test was then applied between groups. RESULTS: TIMP-1 levels were significantly raised in dcSSc and lcSSc groups compared with the RP group and normal controls (p<0.01 to p<0.001). In the dcSSc group, TIMP-1 levels were significantly higher in early disease (<2 years) than in late stage disease (>4 years) (p<0.05). This was not found for the lcSSc group. Serum TIMP-2 and MMP-1 levels in dcSSc and lcSSc did not differ significantly from those in normal controls. Increased levels of TIMPs were not convincingly associated with organ disease. No assay result correlated with autoantibody status (anti-topoisomerase 1 (anti-Scl-70), anticentromere antibody, or anti-RNA polymerase). No significant differences in serum TIMP-1, TIMP-2, or MMP-1 levels were shown in the RP group compared with normal controls. CONCLUSIONS: Raised TIMP-1 levels in the SSc groups support the hypothesis that matrix accumulation occurs in SSc at least in part owing to decreased degradation. Moreover, the variation in TIMP-1 levels between the early and late disease stages of dcSSc seems to reflect the early progressive course of dermal fibrosis seen clinically. The expected reduction in serum MMP-1 levels in the SSc groups was not found. This suggests that tissue matrix accumulation is due to increased inhibitors rather than to decreased MMPs. PMID- 11502612 TI - Association of occupational physical demands and psychosocial working environment with disabling shoulder pain. AB - OBJECTIVE: To estimate the individual and combined associations of physical and psychosocial working environment with disabling shoulder pain and to identify groups at "high risk" for shoulder pain. METHODS: A cross sectional survey was conducted at five manual occupational settings in south Manchester, United Kingdom (n=775, 83%). RESULTS: Both the duration of occupational physical demands (working postures, manual handling activities, and repetitive arm movements) and psychosocial working environment (psychological demands and lack of opportunity to learn new skills) were found to be significantly associated with shoulder pain. Three occupational factors identified a high risk group for shoulder pain: duration of lifting with one hand (prevalence rate ratio (PRR) (highest third) 2.0, 95% confidence interval (CI) 1.4 to 2.8), duration of working above shoulder level (PRR (highest third) 2.2, 95% CI 1.5 to 3.3), and whether employees found their work stressful (PRR (highest third) 1.4, 95% CI 1.0 to 2.1). In addition, a measure of psychological distress (General Health Questionnaire (GHQ) score) was found to identify a group at high risk for shoulder pain (PRR (highest third) 1.9, 95% CI 1.3 to 2.9). In employees exposed to three or more of these factors, 79% (23/29) reported shoulder pain compared with only 16% (56/353) of those not exposed to any. CONCLUSION: This study has identified a variety of occupational physical demands and psychosocial factors associated with shoulder pain. It has also identified groups of employees at a "high risk" for shoulder pain by their exposure to both physical and psychosocial factors. PMID- 11502613 TI - What is the fate of erosions in early rheumatoid arthritis? Tracking individual lesions using x rays and magnetic resonance imaging over the first two years of disease. AB - OBJECTIVES: To investigate the progression of erosions at sites within the carpus, in patients with early rheumatoid arthritis (RA), using magnetic resonance imaging (MRI) and plain radiology over a two year period. METHODS: Gadolinium enhanced MRI scans of the dominant wrist were performed in 42 patients with RA at baseline (within six months of symptom onset) and one year. Plain wrist radiographs (x rays) and clinical data were obtained at baseline, one year, and two years. Erosions were scored by two musculoskeletal radiologists on MRI and x ray at 15 sites in the wrist. A patient centred analysis was used to evaluate the prognostic value of a baseline MRI scan. A lesion centred analysis was used to track the progression of individual erosions over two years. RESULTS: The baseline MRI erosion score was predictive of x ray erosion score at two years (p=0.004). Patients with a "total MRI score" (erosion, bone oedema, synovitis, and tendonitis) > or =13 at baseline were significantly more likely to develop erosions on x ray at two years (odds ratio 13.4, 95% CI 2.65 to 60.5, p=0.002). Baseline wrist MRI has a sensitivity of 80%, a specificity of 76%, a positive predictive value of 67%, and a high negative predictive value of 86% for the prediction of wrist x ray erosions at two years. A lesion centred analysis, which included erosions scored by one or both radiologists, showed that 84% of baseline MRI erosions were still present at one year. When a more stringent analysis was used which required complete concordance between radiologists, all baseline lesions persisted at one year. The number of MRI erosion sites in each patient increased from 2.1 (SD 2.7) to 5.0 (4.6) (p<0.0001) over the first year of disease. When MRI erosion sites were tracked, 21% and 26% were observed on x ray, one and two years later. A high baseline MRI synovitis score, Ritchie score, and erythrocyte sedimentation rate were predictive of progression of MRI erosions to x ray erosions over one year (p=0.005, 0.01, and 0.03 respectively), but there was no association with the shared epitope. Progression of MRI erosions to x ray erosions was not seen in those with transient polyarthritis. CONCLUSIONS: MRI scans of the wrist, taken when patients first present with RA, can predict radiographic erosions at two years. MRI may have a role in the assessment of disease prognosis and selection of patients for more or less aggressive treatment. However, only one in four MRI erosions progresses to an x ray erosion over one year, possibly owing to healing, observer error, or technical limitations of radiography at the carpus. Progression of MRI erosions to x ray erosions is greatest in those with high baseline disease activity. PMID- 11502614 TI - Effect of patient education on adherence to drug treatment for rheumatoid arthritis: a randomised controlled trial. AB - OBJECTIVE: To determine whether a patient education programme (PE) would improve rates of adherence to a slow acting antirheumatic drug and to assess any subsequent effect on patient outcome. METHODS: A randomly controlled study comprising 100 patients with rheumatoid arthritis (49 control CG; 51 experimental EG) requiring D-penicillamine (DPA). The same practitioner saw patients on seven occasions, for the same length of time. The EG received 7 x 30 minute one to one sessions of PE, while the CG received standard management. The primary measure of adherence was a pharmacological marker (phenobarbitone) encapsulated with the DPA assayed at monthly intervals for six months. Plasma viscosity (PV), C reactive protein, articular index, morning stiffness, and pain score were used to assess outcome. RESULTS: 454 blood samples were collected and assayed and the pharmacological marker showed the EG to be significantly more adherent on more occasions than the CG (p<0.05). Patterns of adherence over time showed that at 12 weeks 86% (38/44) of those in the EG compared with 64% (29/45) of the CG remained adherent (p=0.01). These trends continued and by the end of the study 85% (29/34) of the EG compared with 55% (23/42) of the CG were taking their DPA as prescribed. Fifteen patients (12 from the EG) experienced side effects requiring study withdrawal and 14 patients requested study withdrawal (two from the EG). On study entry patients in the CG had significantly higher levels of PV than the EG and this remained so throughout the research. However, on completion, the health status of patients in both groups had improved significantly (p<0.01). CONCLUSIONS: PE significantly increased adherence to DPA and its effects persisted over a period of six months. No additional clinical benefit was detected in the EG in comparison with the CG. PMID- 11502615 TI - Involvement of the peripheral nervous system in primary Sjogren's syndrome. AB - BACKGROUND: Involvement of the peripheral nervous system in patients with primary Sjogren's syndrome (SS) has been reported, but its prevalence in neurologically asymptomatic patients is not well known. OBJECTIVE: To assess clinical and neurophysiological features of the peripheral nervous system in patients with primary SS. PATIENTS AND METHODS: 39 (38 female) consecutive patients with primary SS, aged 20-81 years (mean 50), with a disease duration of 1-30 years (mean 8) were studied. The peripheral nervous system was evaluated by a questionnaire, physical examination, quantified sensory neurological examination, and neurophysiological measurements (nerve conduction studies). To assess autonomic cardiovascular function an orthostatic challenge test, a Valsalva manoeuvre, a forced respiration test, and pupillography were done. RESULTS: Abnormalities as indicated in the questionnaire were found in 8/39 (21%) patients, while an abnormal neurological examination was found in 7/39 (18%) patients. Abnormalities in quantified sensory neurological examination were found in 22/38 (58%) patients. In 9/39 (23%) patients, neurophysiological signs compatible with a sensory polyneuropathy were found. No differences were found in the autonomic test results, disease duration, serological parameters, or erythrocyte sedimentation rate between the patients with primary SS with and those without evidence of peripheral nervous involvement. CONCLUSION: Subclinical abnormalities of the peripheral nervous system may occur in patients with primary SS selected from a department of rheumatology, but clinically relevant involvement of the peripheral nervous system in this patient group is rare. PMID- 11502617 TI - A pilot study of a long acting somatostatin analogue for the treatment of refractory rheumatoid arthritis. AB - OBJECTIVE: To evaluate the efficacy and safety of a long acting somatostatin analogue in a subset of patients with refractory rheumatoid arthritis (RA). METHODS: Ten patients with active, refractory RA, who had failed to respond to at least four disease modifying antirheumatic drugs (DMARDs), were treated with monthly intramuscular injections of 20 mg of a long acting preparation of octreotide (Sandostatin-LAR) for three months. They were evaluated every two weeks in an open label pilot study. The primary measure of clinical response was the American College of Rheumatology criteria for a 20% improvement in measures of disease activity (ACR 20). RESULTS: Eight patients completed the 14 week trial, while two patients received only one or two doses of the somatostatin analogue, but were eligible for evaluation. On an intention to treat basis 6/10 patients responded: four patients met the ACR 20 criteria at weeks 6-10, while two patients continued to improve with time, and met the ACR 50 and 70 criteria respectively, at week 14. On evaluation of the 10 patients as a group, a significant improvement (p<0.05) was noted in the mean visual analogue scales of pain, doctor's and patient's global assessment of disease activity, and in the mean number of swollen joints. Adverse effects were minor: transient bloating and loose stools, an urticarial rash (n=1), and a transient increase of liver enzymes (n=1). CONCLUSION: Treatment with a long acting somatostatin analogue led to significant clinical improvement in a subset of patients with active, refractory RA. The treatment was relatively safe and well tolerated. Further large, placebo controlled studies are required to evaluate this drug as a potential DMARD for patients with RA. PMID- 11502616 TI - Performance of two ELISAs for antifilaggrin autoantibodies, using either affinity purified or deiminated recombinant human filaggrin, in the diagnosis of rheumatoid arthritis. AB - OBJECTIVE: To develop a standardisable enzyme linked immunosorbent assay (ELISA), using human filaggrin, for detection of antifilaggrin autoantibodies in rheumatoid arthritis (RA). To compare the diagnostic performance of the ELISA with those of reference tests: "antikeratin antibodies" ("AKA"), and antibodies to human epidermis filaggrin detected by immunoblotting (AhFA-IB). METHODS: Two ELISAs were developed using either affinity purified neutral-acidic human epidermis filaggrin (AhFA-ELISA-pur) or a recombinant human filaggrin deiminated in vitro (AhFA-ELISA-rec) as immunosorbent. Antifilaggrin autoantibodies were assayed in 714 serum samples from patients with well characterised rheumatic diseases, including 241 RA and 473 other rheumatic diseases, using the two ELISAs. "AKA" and AhFA-IB tests were carried out in the same series of patients. The diagnostic performance of the four tests was compared and their relationships analysed. RESULTS: The titres of "AKA", AhFA-IB, and the AhFA-ELISAs correlated strongly with each other. The diagnostic sensitivity of the AhFA-ELISA-rec, which was better than that of AhFA-ELISA-pur, was 0.52 for a specificity of 0.95. This performance was similar to those of "AKA" or AhFA-IB. However, combining AhFA ELISA-rec with AhFA-IB led to a diagnostic sensitivity of 0.55 for a specificity of 0.99. CONCLUSION: A simple and easily standardisable ELISA for detection of antifilaggrin autoantibodies was developed and validated on a large series of patients using a citrullinated recombinant human filaggrin. The diagnostic performance of the test was similar to that of the "AKA" and AhFA-IB. Nevertheless, combining the AhFA-ELISA-rec with one of the other tests clearly enhanced the performance. PMID- 11502618 TI - Lymphoma in a patient with rheumatoid arthritis receiving methotrexate treatment: successful treatment with rituximab. AB - A 55 year old man with chronic lymphocytic leukaemia (CLL) and rheumatoid arthritis (RA), treated for four years with methotrexate (MTX), who developed a B cell non-Hodgkin's lymphoma (B-NHL), is described. The tumour was localised to the shoulder and axillary lymph nodes, and positive for Epstein-Barr viral antigens. After failure of radiation and chemotherapy, a complete remission was achieved with a combination of antibody treatment (rituximab) and EPOCH. The development of a second malignancy in a patient with RA receiving MTX has not been described before. The summation of T cell deficiencies induced by MTX, CLL, and RA may all have contributed to the development of the B-NHL. PMID- 11502619 TI - Benign familial hypermobility syndrome and Trendelenburg sign in a painting "The Three Graces" by Peter Paul Rubens (1577-1640). AB - Clinical features suggestive of hypermobility syndrome and a positive Trendelenburg sign are described in a painting "The Three Graces" (1638-1640) by Peter Paul Rubens, Prado, Madrid. The most obvious findings are scoliosis, positive Trendelenburg sign, and hyperextension of the metacarpal joints, hyperlordosis, and flat feet. The sitters, presumably Helene Fourment (second wife of Rubens) and her sisters, support the hereditary familial aspect of hypermobility. Manifest hypermobility of the hand has also been found in two other ancient paintings: "Saint Cyriaque" in the Heller Retable by Mathias Grunewald (1450-1528), Frankfurt, and "The wounded man" by Gaspare Traversi, Venice (1732-1769). The finding of signs of hypermobility in ancient paintings shows that artists who are keen observers of nature could describe, or at least record, this condition long before doctors did. The art of the past can be a useful tool in the field of paleopathology. PMID- 11502620 TI - Committee on Manpower of Pulmonary and Critical Care Societies: a report to membership. PMID- 11502621 TI - Lest the tide return. PMID- 11502622 TI - Smear-negative pulmonary tuberculosis in industrialized countries. PMID- 11502623 TI - Adenosine deaminase in the diagnosis of tuberculous pleural effusion. PMID- 11502624 TI - The obesity-hypoventilation syndrome revisited. PMID- 11502625 TI - Noninvasive cardiac output monitoring. PMID- 11502626 TI - Enhancing patient safety for pediatric bronchoscopy: alternatives to conscious sedation. PMID- 11502627 TI - Treatment experience of multidrug-resistant tuberculosis in Florida, 1994-1997. AB - OBJECTIVE: To study the clinical characteristics and results of patients with diagnoses of multidrug-resistant tuberculosis (MDR-TB) in the state of Florida. METHODS: Retrospective chart review of all patients (n = 81) with diagnoses of MDR-TB in Florida between January 1, 1994, and July 31, 1997. RESULTS: The average number of resistant drugs was 4.8 (range, 2 to 11). Of 81 patients, 46 patients (57%) completed adequate therapy, 26 patients (32%) died, and 9 patients (11%) never completed a satisfactory course of treatment. Patients who received at least part of their therapy at A. G. Holley State Hospital, a specialized tuberculosis (TB) treatment center, had significantly higher treatment completion rates (79%) than those treated as outpatients alone (48% treatment completion rate, p < 0.001), even after the exclusion of patients who were acutely ill and died within 2 months of diagnosis. CONCLUSION: In Florida, a specialized TB care program for MDR-TB, including at least partial inpatient therapy, yielded higher treatment completion rates compared to outpatient treatment alone. PMID- 11502628 TI - Identifying pulmonary tuberculosis in patients with negative sputum smear results. AB - BACKGROUND: Clinicians need to decide whether to begin empiric therapy for patients who are suspected of having tuberculosis (TB) but have negative sputum smear results. Culture results may take weeks, and delaying treatment may allow further transmission of disease. STUDY OBJECTIVE: To identify the clinical, demographic, and radiographic characteristics that identify smear-negative patients who have TB, and to create a TB prediction rule. DESIGN: Retrospective chart review. SETTING: University-affiliated public hospital in San Francisco, CA, between 1993 and 1998. PATIENTS: Forty-seven patients with TB and 141 control patients who were hospitalized with a suspicion of pulmonary TB; all had negative sputum smear results. MEASUREMENTS AND RESULTS: Demographic, clinical, and radiographic variables were determined by chart review. In multivariate analysis, a positive tuberculin skin test result (odds ratio [OR], 4.8; 95% confidence interval [CI], 2.0 to 11.9) was independently associated with an increased risk of a positive TB culture finding. A radiographic pattern not typical of pulmonary tuberculosis (OR, 0.3; 95% CI, 0.1 to 0.7) and expectoration with cough (OR, 0.3; 95% CI, 0.1 to 0.6) were predictive of a decreased risk. An interaction between HIV seropositivity and mediastinal lymphadenopathy on the chest radiograph was also associated with a positive TB culture result (OR, 7.2; 95% CI, 1.4 to 36.0). The TB prediction score (TPS) was created with widely ranging likelihood ratios that could affect the posterior probability of TB by 30-fold. CONCLUSION: The TPS put into context with the overall prevalence of TB in a given area may help clinicians decide if a patient with negative sputum smear results should start empiric antituberculous therapy or wait for culture results. These results need prospective validation. PMID- 11502629 TI - Adenosine deaminase levels in nontuberculous lymphocytic pleural effusions. AB - OBJECTIVES: Adenosine deaminase (ADA) can aid in the diagnosis of tuberculous pleural effusions, but false-positive findings from lymphocytic effusions have been reported. We studied the ADA levels in a variety of nontuberculous lymphocytic effusions and analyzed the relationships between ADA and conventional hematologic and biochemical parameters. METHODS: One hundred six lymphocytic pleural fluid samples (lymphocyte count > 50%) were analyzed. These included post coronary artery bypass grafting (CABG) effusions (n = 45), malignant effusions (n = 27), miscellaneous exudative effusions (n = 10), and transudative effusions (n = 24). ADA levels were determined using the Giusti method. In 22 randomly selected cases, ADA was measured again on the same sample 6 weeks later. RESULTS: The ADA level reached the diagnostic cutoff for tuberculosis (40 U/L) in only three cases (2.8%): two lymphomas and one complicated parapneumonic effusion. There was no significant correlation between effusion ADA levels and the total leukocyte (r = 0.08), differential lymphocyte (r = 0.18) or monocyte (r = - 0.18) counts. ADA levels were significantly lower in the transudative effusions (7.2 +/ 3.5 U/L) than in post-CABG (16.6 +/- 7.2 U/L), malignant (15.3 +/- 11.2 U/L), and other exudative (15.4 +/- 13.1 U/L) effusions (p < 0.001). ADA measurements were consistent when assayed 6 weeks apart (r = 0.95; p < 0.00001; coefficient of variation, 14%). CONCLUSIONS: ADA levels in nontuberculous lymphocytic effusions seldom exceeded the diagnostic cutoff for TB. Effusion ADA levels cannot be predicted from total or differential leukocyte counts. Post-CABG pleural fluids had ADA levels similar to other nontuberculous lymphocytic effusions. ADA is stable in effusion fluids, and its measurement is reproducible. PMID- 11502630 TI - Soluble leukocyte selectin in the analysis of pleural effusions. AB - STUDY OBJECTIVES: To determine if soluble leukocyte selectin (sL-selectin) levels in serum and pleural fluid (PF) are an inflammatory marker that differentiates pleural effusion transudates from exudates. DESIGN: sL-selectin PF and serum levels were measured in consecutive patients and compared to established criteria. SETTING: A tertiary-care military medical center. PATIENTS: One hundred twenty patients undergoing diagnostic or therapeutic thoracentesis. INTERVENTIONS: PF and serum samples were collected during thoracentesis and analyzed separately for sL-selectin levels. Results were compared with clinical diagnosis and established PF criteria including the criteria of Light et al, cholesterol ratio, total bilirubin ratio, and albumin gradient. MEASUREMENTS AND RESULTS: sL-selectin levels in PF and serum were determined in 109 patients. By clinical diagnosis, mean +/- SD PF sL-selectin levels were 200.2 +/- 124.3 ng/mL in transudates and 496.8 +/- 379.2 ng/mL in exudates (p < 0.001). By the criteria of Light et al, mean PF sL-selectin levels were 195.7 +/- 105.2 ng/mL in transudates and 448.2 +/- 367.6 ng/mL in exudates (p < 0.001). Mean sL-selectin PF to serum ratios were 0.31 +/- 0.17 in transudates and 0.72 +/- 0.31 in exudates (p < 0.001) by clinical criteria, and 0.31 +/- 0.18 in transudates and 0.64 +/- 0.33 in exudates (p < 0.001) by the criteria of Light et al. No significant difference was noted with serum sL-selectin levels between groups. CONCLUSIONS: sL-selectin is an inflammatory marker that differentiates transudates from exudates in pleural effusions and is a sensitive indicator for PF analysis. PMID- 11502631 TI - The obesity-hypoventilation syndrome revisited: a prospective study of 34 consecutive cases. AB - STUDY OBJECTIVES: Obesity has many detrimental effects on the respiratory function and may lead to chronic hypoventilation in some patients, an association known as the obesity-hypoventilation syndrome (OHS). In many cases, patients with OHS also have sleep apneas. Hereafter, we describe several features of a cohort (n = 34) of patients with OHS and show the comparisons with a large cohort (n = 220) of patients with obstructive sleep apnea syndrome (OSAS). We compare also OHS patients with a group of patients with the association of OSAS and COPD, also known as "overlap" patients. DESIGN: Descriptive analysis of prospectively collected clinical data. SETTING: Respiratory care unit and sleep laboratory of university hospital. RESULTS: In OHS patients, OSAS was present in most of the cases (23 of 26 patients). However, in three patients, OHS was not associated with OSAS, showing that obesity per se may lead to chronic hypoventilation. As expected by definition, OHS patients had, on average the worst diurnal arterial blood gas measurements, compared to the other groups. For the OHS patients, the mean diurnal PaO(2) was 59 +/- 7 mm Hg, which was significantly different from the PaO(2) of the OSAS patients (75 +/- 10 mm Hg; p = 0,001) but also from the overlap patients (66 +/- 10 mm Hg; p = 0.015). Pulmonary hypertension (ie, mean pulmonary artery pressure > 20 mm Hg) was more frequent in OHS patients than in "pure" OSAS patients (58% vs 9%; p = 0.001). CONCLUSION: Patients with OSAS and chronic respiratory insufficiency had in most cases an associated OHS or COPD. Patients with OHS were older than patients with pure OSAS. They had mild-to moderate degrees of restrictive ventilatory pattern due to obesity. Severe gas exchange impairment and pulmonary hypertension were quite frequent. The association of OHS and OSAS was the rule. However, in three patients, OHS was not associated with OSAS, suggesting that OHS is an autonomous disease. PMID- 11502633 TI - Home oximetry studies for diagnosis of sleep apnea/hypopnea syndrome: limitation of memory storage capabilities. AB - BACKGROUND: Memory oximeters enable diagnostic studies for sleep apnea hypopnea syndrome (SAHS) to be performed in the home. However, memory capabilities may be limited. STUDY OBJECTIVES: To compare a pulse oximeter used at home with an 8-h memory, storing data every 12 s, and in the laboratory, with on-line recording every 2 s. DESIGN: Prospective cohort study. SETTING: Patients' homes and a sleep laboratory. PATIENTS: One hundred patients with suspected SAHS. MEASUREMENTS: Home oximetry and a laboratory full polysomnography. The number of >/= 4% dips in pulse oximetric saturation (SpO(2)) was calculated for each study. Daytime sleepiness was assessed by the Epworth Sleepiness Scale (ESS) score. RESULTS: The mean dips per hour were 5.3/h (range, 0 to 53/h) for home studies and 13.4/h (range, 0 to 106/h) for laboratory studies; the relationship between home and laboratory studies was as follows: home = (0.4 x laboratory) - 0.01 +/- 11.2; r(2) = 0.64. Mean difference was 8.4/h (- 2.5 to + 77.9/h), which correlated with the mean of the measurements. At a cutoff point of 10/h, 52 studies were both negative and 13 studies were both positive. Nineteen home studies were false negatives. Sensitivity was 0.41, and specificity was 1.0. In these 19 studies, 7 patients had an ESS score > 10 and 4 patients had an ESS score > 14. To confirm that differences were due to different sampling rates, 16 additional patients had on-line data and stored data collected simultaneously in the laboratory. Mean dips per hour were 3.2/h (range, 0.1 to 18.3/h) for the stored data and 8.34/h (0.2 to 22.8/h) for on-line data; the relationship being stored was as follows: 0.5 on-line - 1.17 +/- 2.6; r(2) = 0.69. Mean difference was 5.2/h (0.04 to 15.4 h), which correlated with the mean of the measurements. CONCLUSION: Home studies using a memory storage pulse oximeter may underestimate the number of hypoxic dips, probably due to sampling rates. Clinically significant hypoxic SAHS may therefore be missed. PMID- 11502632 TI - The use of health-care resources in obesity-hypoventilation syndrome. AB - OBJECTIVE: To document health-care utilization (ie, physician claims and hospitalizations) in patients with obesity-hypoventilation syndrome (OHS), for 5 years prior to the diagnosis and for 2 years after the diagnosis and initiation of treatment. DESIGN: Retrospective observational cohort study. SETTING: University-based sleep disorders center in Manitoba, Canada. PATIENTS AND CONTROL SUBJECTS: Twenty OHS patients (mean [+/- SD] age, 52.7 +/- 9.5 years; body mass index [BMI], 47.3 +/- 11.0 kg/m(2); PaCO(2), 59.7 +/- 13.8 mm Hg; PaO(2), 51.6 +/ 12.4 mm Hg) were matched to two sets of control subjects. First, each case was matched to 15 general population control subjects (GPCs) by age, gender, and geographic location, and, second, each case was matched to a single obese control subject (OBC) who was matched using the same criteria as for the GPCs, plus the measurement of BMI. MEASUREMENTS AND RESULTS: In the 5 years before diagnosis, the 20 OHS patients had (mean +/- SE) 11.2 +/- 1.8 physician visits per patient per year vs 5.7 +/- 0.8 (p < 0.01) visits for OBCs and 4.5 +/- 0.4 (p < 0.001) visits for GPCs. OHS patients generated higher fees, $623 +/- 96 per patient per year for the 5 years prior to diagnosis compared to $252 +/- 34 (p < 0.001) for OBCs and $236 +/- 25 (p < 0.001) for GPCs. OHS patients were much more likely to be hospitalized than were subjects in either control group in the 5 years prior to diagnosis (odds ratio [OR] vs GPCs, 8.6) (95% confidence interval [CI], 5.9 to 12.7); OR vs OBCs, 4.9 (95% CI, 2.3 to 10.1). In the 2 years after diagnosis and the initiation of treatment (usually continuous positive airway pressure or bilevel positive airway pressure), there was a significant linear reduction in physician fees. In the 2 years after the initiation of treatment, there was a 68.4% decrease in days hospitalized per year (5 years before treatment, 7.9 days per patient per year; after 2 years of treatment, 2.5 days per patient per year [p = 0.01]). CONCLUSIONS: OHS patients are heavy users of health care for several years prior to evaluation and treatment of their sleep breathing disorder; there is a substantial reduction in days hospitalized once the diagnosis is made and treatment is instituted. PMID- 11502634 TI - Abdominal muscle activity in sleep apnea during continuous positive airway pressure titration. AB - STUDY OBJECTIVES: The aim of this study was to investigate whether presence of expiratory abdominal muscle activity (EAMA) in obstructive sleep apnea syndrome (OSAS) patients during nasal continuous positive airway pressure (nCPAP) is due to either nCPAP overprescription or nCPAP underprescription. DESIGN: Airflow, esophageal pressure (Pes), and gastric pressure (Pga) were routinely measured during polysomnography aimed at determining the optimal nCPAP level, and the magnitude of EAMA was evaluated in relation to the nCPAP level and to the conventional indexes of upper-airway obstruction used during nCPAP titration. PATIENTS: The study was performed 12 patients with OSAS. RESULTS: Six patients displayed sustained EAMA, ie, EAMA lasting > 3 min, and characterized by a decrease in abdominal diameter and a paradoxical rise in Pga during expiration. In all six patients, EAMA decreased gradually as nCPAP neared optimal levels, and then disappeared when the optimal nCPAP level was achieved. The decrease in EAMA as nCPAP increased was associated with an increase in minute ventilation, decreases in both inspiratory and expiratory resistance, a decrease in Pes swing, and the normalization of the inspiratory flow contour. CONCLUSIONS: We conclude that the EAMA observed in some OSAS patients might be an indirect marker of upper airway obstruction, and that the presence of EAMA during nCPAP titration might indicate a suboptimal nCPAP level rather than a deleterious effect of nCPAP. PMID- 11502635 TI - Combined effects of a nasal dilator and nasal prongs on nasal airflow resistance. AB - STUDY OBJECTIVES: Nasal prongs (NPs), when used to assess nasal flow, can result in dramatic increases in nasal airflow resistance (NR). The aim of this study was to investigate whether the NP-induced increases in NR could be corrected by the simultaneous use of an internal nasal dilator (ND). DESIGN: NR was estimated by posterior rhinomanometry, in the basal state (NRb), and while breathing with NP (NRp), with ND (NRd), and with both ND and NP (NRd + p). PARTICIPANTS: The study was performed in 15 healthy subjects. MEASUREMENTS AND RESULTS: NR (mean NRb [+/- SEM], 2.5 +/- 0.4 cm H(2)O/L/s) significantly decreased with ND (NRd = 1.4 +/- 0.2 cm H(2)O/L/s; p < 0.001) and significantly increased with NP (NRp = 3.8 +/- 0.8 cm H(2)O/L/s; p < 0.001). A significant logarithmic relationship was found between NRd and NRb (r(2) = 0.95; p < 0.0001), and a significant exponential relationship was found between NRp and NRb (r(2) = 0.99; p < 0.0001). While breathing with both ND and NP, NRd + p was significantly lower than NRb (1.9 +/- 1.4 cm H(2)O/L/s; p < 0.02). CONCLUSIONS: Our results demonstrate that the ND tends to slightly overcorrect the NP-induced increase in NR and suggest that, in view of the possible effects of NPs on upper airway resistance, the combination of both devices might be used for nasal airflow monitoring during nocturnal polysomnography in patients presenting with highly resistive nares. PMID- 11502636 TI - Perception of dyspnea in patients with neuromuscular disease. AB - BACKGROUND: The perception of dyspnea is not a prominent complaint of resting patients with neuromuscular disease (NMD). To our knowledge, no study has been addressed at evaluating the interrelationships among lung mechanics, respiratory motor output, and the perception of dyspnea in patients with NMD receiving ventilatory stimulation. MATERIAL: Eleven patients with NMD (mean +/- SD age, 44 +/- 11.8 years; 5 men) of different etiology and a group of normal subjects matched for age and sex (control subjects). METHODS: While patients were breathing room air, lung volumes, arterial blood gases, the pattern of breathing (minute ventilation [E], tidal volume [VT], respiratory frequency, inspiratory time), and maximal (less negative) esophageal pressure during a sniff maneuver (Pessn), as an index of inspiratory muscle strength, were measured. Then we evaluated the response to hypercapnic-hyperoxic stimulation (hypercapnic hyperoxic rebreathing test [RT]) in terms of breathing pattern, inspiratory swing of pleural pressure (Pessw), and inspiratory effort (Pessw[%Pessn]). During the RT, dyspnea was assessed every 30 s using a modified Borg scale (0 to 10). RESULTS: Pulmonary volumes were reduced in seven patients, and PCO(2) was out of proportion to E in four patients. Group Pessn was 42.8 +/- 23.6 cm H(2)O in patients and 107 +/- 20.4 cm H(2)O in control subjects (p < 0.001). Dynamic elastance (Eldyn) [p = 0.0016] and Pessw(%Pessn) [p < 0.0005] were higher in patients. During the RT, Borg/CO(2), Pessw(%Pessn)/CO(2), and Borg/Pessw(%Pessn) were similar in the two groups, while E/CO(2) and VT/CO(2) were lower in patients (p < 0.0002 for both). As a consequence, for unit change in VT (percentage of predicted vital capacity [%VC]), greater changes in Pessw(%Pessn) were associated with greater Borg scores in patients. Baseline Eldyn related to Pessw(%Pessn)/VT(%VC) during hypercapnia (r(2) = 0.85), an index of neuroventilatory coupling of the ventilatory pump (NVC). NVC predicted a good amount of the variability in Borg/E (r(2) = 0.46, p < 0.02). CONCLUSIONS: In this subset of NMD patients during hypercapnic stimulation, a normal inspiratory motor output per unit change in PCO(2) results in a shallow breathing pattern. The consequent impairment of NVC underlies the higher scoring of dyspnea in these patients. PMID- 11502637 TI - The perception of dyspnea in patients with mild asthma. AB - BACKGROUND: Airway function, as assessed by standard spirometry, and the intensity of dyspnea reported by asthmatic patients correlate poorly. OBJECTIVE: This study tests the following two hypotheses: (1) that measures of the tendency of a patient to somatize will reduce the variation in the report of dyspnea not explained by airway function; and (2) that plethysmography is a better tool with which to estimate the degree of dyspnea associated with asthma. DESIGN: A prospective laboratory study carried out over one study session. PARTICIPANTS: Forty asthmatic subjects who had withheld bronchodilator (BD) therapy overnight. INTERVENTIONS: We performed spirometry, plethysmography, and an assessment of dyspnea (ie, modified Borg scale) on all subjects before and after they received BD therapy. Standard questionnaires pertaining to psychological state and trait were administered as well. RESULTS: The change in specific airway conductance with BD therapy correlated with a decline in the Borg score (r = 0.47; p = 0.007). By contrast, neither spirographic measures nor measures of static lung volumes correlated. Correlation with the Borg scale score was not improved by adding indexes of either somatization or psychological state or trait. CONCLUSION: The relief of dyspnea reported by patients with mild asthma after BD therapy is related to dilatation of the central airways. PMID- 11502638 TI - Evaluation of blood vessels and edema in the airways of asthma patients: regulation with clarithromycin treatment. AB - BACKGROUND: Although airway angiogenesis and edema have been proposed to contribute to the airway remodeling process in patients with asthma, there are few studies looking at these structural components in the airway tissue of asthma patients. Mycoplasma infection may be associated with chronic asthma and has been shown to induce angiogenesis and edema in a murine model. PARTICIPANTS AND MEASUREMENTS: We evaluated blood vessels and edema by immunohistochemistry in endobronchial biopsy samples from 10 normal control subjects and 15 patients with mild-to-moderate asthma before and after a 6-week treatment with clarithromycin (n = 8) or placebo (n = 7). Type IV collagen and alpha(2)-macroglobulin were used to identify blood vessels and edema in the tissue, respectively. Mycoplasma pneumoniae was evaluated by polymerase chain reaction. SETTING: National Jewish Medical and Research Center. RESULTS: At baseline, the vascularity, the number of blood vessels, and the edematous area in the airway tissue were not significantly different between asthmatic patients and normal control subjects. However, asthmatic patients demonstrated increased blood vessel size compared with normal control subjects (p = 0.03). After clarithromycin treatment in asthmatic patients, the number of blood vessels was increased (p = 0.02), while edema decreased (p = 0.049). Asthmatic patients who tested positive for M pneumoniae showed a significant increase in vascularity than asthmatic patients who tested negative for M pneumoniae (p = 0.02). CONCLUSION: Our data suggest that angiogenesis and edema may not be significant features of airway remodeling in patients with chronic, mild-to-moderate asthma. Clarithromycin treatment in asthmatic patients could reduce the edematous area as identified by alpha(2) macroglobulin staining, which may lead to airway tissue shrinkage and cause an artificial increase in the number of blood vessels. PMID- 11502639 TI - Salmeterol powder provides significantly better benefit than montelukast in asthmatic patients receiving concomitant inhaled corticosteroid therapy. AB - STUDY OBJECTIVES: Comparison of inhaled salmeterol powder vs oral montelukast treatment in patients with persistent asthma who remained symptomatic while receiving inhaled corticosteroids. DESIGN: Randomized, double-blind, double dummy, parallel-group, multicenter trials of 12-week duration. SETTING: Outpatients in private and university-affiliated clinics. PATIENTS: Male and female patients > or = 15 years of age with a diagnosis of asthma (baseline FEV(1) of 50 to 80% of predicted) and symptomatic despite receiving inhaled corticosteroids. INTERVENTIONS: Inhaled salmeterol xinafoate powder, 50 microg bid, or oral montelukast, 10 mg qd. MEASUREMENTS AND RESULTS: Treatment with salmeterol powder resulted in significantly greater improvements from baseline compared with montelukast for most efficacy measurements, including morning peak expiratory flow (35.0 L/min vs 21.7 L/min; p < 0.001), percentage of symptom-free days (24% vs 16%; p < 0.001), and the percentage of rescue-free days (27% vs 20%; p = 0.002). Total supplemental albuterol use was decreased significantly more in the salmeterol group compared with the montelukast group (- 1.90 puffs per day vs - 1.66 puffs per day; p = 0.004) and nighttime awakenings per week decreased significantly more with salmeterol than with montelukast (- 1.42 vs - 1.32; p = 0.015). Patients treated with inhaled salmeterol were significantly more satisfied with their treatment regimen and how well, how fast, and how long it worked than were patients who were treated with oral montelukast. The safety profiles for the two treatments were similar. CONCLUSION: In patients with persistent asthma who remain symptomatic while receiving inhaled corticosteroids, adding inhaled salmeterol powder provided significantly greater improvement in lung function and asthma symptoms and was preferred by patients over oral montelukast. PMID- 11502640 TI - Electrolyte disturbances in patients with chronic, stable asthma: effect of therapy. AB - OBJECTIVE: To determine the prevalence of electrolyte disturbances in patients with chronic, stable asthma, and to assess whether the therapeutic agents used to treat chronic asthma have an effect on abnormal electrolyte levels. DESIGN: Prospective, hospital-based, cross-sectional study. SETTING: University teaching hospital in Jeddah, Saudi Arabia. PATIENTS: Patients with chronic, stable asthma. METHOD: Ninety-three consecutive patients with chronic, stable asthma were involved in the study. On the day of the visit to the asthma clinic, particulars such as age, sex, duration of asthma, and details of drug therapy were obtained from each asthmatic patient. Serum potassium, magnesium, phosphorus, calcium, and sodium levels were measured. Normal values were as follows: potassium, 3.5 to 5 mmol/L; magnesium, 0.74 to 1.2 mmol/L; phosphorus, 0.8 to 1.4 mmol/L; and calcium, 2.1 to 2.6 mmol/L. RESULTS: Electrolyte disturbances were found in 43% of the patients; 85% of the patients had one electrolyte disturbance, 10% had two electrolyte disturbances, and 5% had three electrolyte disturbances. The highest proportions were for magnesium (26.9%) and phosphorus (15.1%) [serum levels were 0.69 +/- 0.04 mmol/L and 0.64 +/- 0.09 mmol/L, respectively], the lowest proportions were for potassium (5.4%) and sodium (4.3%) [serum levels were 3.3 +/ 0.01 mmol/L and 133 +/- 0.01 mmol/L, respectively], and no patient had a calcium disturbance. Logistic regression analysis showed no statistically significant association between the therapy used and electrolyte disturbances. CONCLUSION: Hypomagnesemia and hypophosphatemia were found to be the two most common electrolyte disturbances in patients with chronic, stable asthma. Therapeutic agents used to treat patients with chronic asthma have no effect on abnormal electrolyte levels. The underlying cause still remains unclear. PMID- 11502641 TI - Effects of supplemental oxygen during activity in patients with advanced COPD without severe resting hypoxemia. AB - STUDY OBJECTIVES: To assess oxygen desaturation during activities and to evaluate the short-term effects of supplemental O(2) use in patients with severe COPD who do not qualify for long-term O(2) therapy. DESIGN: A double-blind, randomized, placebo-controlled trial. SETTING: Outpatients from the pulmonary diseases division of a tertiary-care university hospital. PATIENTS: Twenty patients with stable COPD with FEV(1)/FVC ratios of < 50%, FEV(1) levels < 55% of the predicted normal value, and PaO(2) levels of > 60 mm Hg when resting. INTERVENTIONS: Patients were initially evaluated with pulmonary function tests, blood gas analysis, and Doppler echocardiography, and they underwent the following three 6 min walking tests (WTs) in a random sequence: basal WT (BWT); WT while breathing compressed air (CAWT); and WT while breathing O(2) (O(2)WT). MEASUREMENTS AND RESULTS: The distance walked was recorded in meters. Dyspnea was measured by Borg scale measurement before and after the tests, and arterial oxygen saturation measured by pulse oximetry (SpO(2)) was continuously monitored. Results were analyzed by grouping patients in the following manner: desaturators (DSs) (ie, patients with a drop in SpO(2) of at least 5% and < 90% during the WT) vs nondesaturators (NDSs); and O(2) responders (ie, patients with an increase of at least 10% in the distance walked and/or a decrease of at least 3 points in Borg index score) vs nonresponders. During the BWT, 11 of 20 patients (55%) were defined as desaturators. During the O(2)WT, the SpO(2) remained at > 90% in every patient. The distance walked increased by 22% (p < 0.02), and dyspnea decreased 36% (p < 0.01) in DS patients. In NDS patients, O(2) administration reduced dyspnea by 47% (p < 0.001), but the distance walked did not improve. Responses were markedly different from one patient to another. No significant differences were noticed between the results of the BWT and CAWT in any of the groups. Thirteen O(2) responders did not differ from 7 nonresponders either in basal data or in desaturation measure during the BWT, except that all walking responders (five patients) were above the median of basal left ventricle performance. CONCLUSIONS: Most of the studied COPD patients desaturated during the BWT. O(2) administration avoided desaturation and could increase the distance walked and reduce dyspnea, but these effects were not related to walking desaturation in individual cases. Improvements were not a placebo effect. The therapeutic role of O(2) during activities in some patients with severe COPD needs to be individually assessed. PMID- 11502642 TI - Anatomic landmarks to estimate the length of the diaphragm from chest radiographs: effects of emphysema and lung volume reduction surgery. AB - STUDY OBJECTIVES: To define anatomic landmarks that can be used to predict insertions of the diaphragm on chest radiographs and to estimate diaphragm length. DESIGN: Prospective clinical trial with a parallel group design. SETTING: Laboratory investigations in normal volunteers recruited by advertisement and in emphysema outpatients being evaluated for elective bilateral lung volume reduction surgery (LVRS). PATIENTS: Twenty-six normal subjects classified into young and older age groups, with a third group of 13 emphysema patients matched for age and sex with the older group. MEASUREMENTS: Identification and between group comparisons were made of anatomic landmarks on anteroposterior and lateral chest radiographs obtained at total lung capacity. Predicted landmarks were generated from normal subjects. Within-subject and between-group comparisons were made of diaphragm length index (DLI) based on observed anatomic landmarks (DLIobs) and diaphragm length index based on predicted anatomic landmarks (DLIpred) at functional residual capacity. RESULTS: Anatomic landmarks were not different between the three groups or between male and female subjects, and were not different before and after LVRS in emphysema patients. No difference was found between DLIobs and DLIpred in normal subjects and emphysema patients, but both were smaller in emphysema patients than in normal subjects and increased after LVRS in emphysema patients. CONCLUSION: This study validates the use of anatomic landmarks to estimate DLI. Using these landmarks simplifies the determination of diaphragmatic lengths and could be a useful tool for the evaluation of the functional capacity of the diaphragm, and possibly as a prognostic indicator of patients who are candidates for LVRS. PMID- 11502643 TI - Effects of vesnarinone on peripheral circulating levels of cytokines and cytokine receptors in patients with heart failure: a report from the Vesnarinone Trial. AB - STUDY OBJECTIVES: Proinflammatory cytokines may contribute to disease progression in heart failure by virtue of the direct toxic effects that these molecules exert on the heart and the circulation. Accordingly, there is interest in developing therapeutic agents with anticytokine properties that might be used as adjunctive therapy to modulate proinflammatory cytokine levels in patients with heart failure. Previous experimental studies suggested that vesnarinone has potent anticytokine properties in vitro. Therefore, we examined the effects of vesnarinone on circulating levels of cytokines and cytokine receptors in a large scale, multicenter, clinical trial of patients with moderate-to-advanced heart failure: the Vesnarinone Trial (VEST). METHODS: Circulating levels of tumor necrosis factor (TNF)-alpha, soluble TNF-receptor type 1, soluble TNF-receptor type 2, as well as interleukin (IL)-6 and soluble IL-6 receptor (sIL-6R) were measured on plasma samples by enzyme-linked immunosorbent assay at baseline and at 24 weeks in patients who were receiving placebo (n = 352), 30 mg of vesnarinone (n = 367), and 60 mg of vesnarinone (n = 327). RESULTS: Treatment with 30 mg and 60 mg of vesnarinone had no effect on circulating levels of cytokines or cytokine receptors in patients with advanced heart failure over a 24 week period. CONCLUSIONS: In contrast to the potent anticytokine effects observed with vesnarinone in experimental studies in vitro, the results of this clinical study suggest that vesnarinone does not have any measurable anticytokine effects in vivo in patients with moderate-to-advanced heart failure. PMID- 11502644 TI - A pilot safety trial of prolonged (48 h) infusion of the dual endothelin-receptor antagonist tezosentan in patients with advanced heart failure. AB - STUDY OBJECTIVES: Tezosentan, an IV dual endothelin-receptor antagonist, has demonstrated beneficial hemodynamic effects in patients with advanced heart failure. In addition, no notable differences in safety and tolerability variables were detected between tezosentan-treated and placebo-treated patients when infused over 4 to 6 h. The present study was conducted primarily to assess the safety and tolerability of tezosentan when administered over a prolonged, 48-h treatment period, and secondarily to investigate hemodynamic response. DESIGN: This randomized, double-blind, active-controlled study of continual IV administration of two dosages of tezosentan (20 mg/h and 50 mg/h; n = 6 each) or dobutamine (5 microg/kg/min; n = 2) over 48 h in patients with advanced heart failure was conducted to assess tolerability, safety, and hemodynamic variables (Doppler echocardiography). RESULTS: During tezosentan infusion, no episodes of hypotension requiring withdrawal of therapy occurred, and hemodynamic rebound was not observed after abrupt cessation of the infusion. There were no reports of worsening heart failure in tezosentan-treated patients up to 28 days following the infusion. The most common side effect during the infusion was headache (9 of 12 tezosentan-treated patients and both dobutamine-treated patients). Echocardiographic Doppler measurements suggested improvements in cardiac index, pulmonary capillary wedge pressure, and relaxation properties as well as in diastolic and systolic function in all treatment groups. CONCLUSIONS: Prolonged, 48-h IV dual endothelin-receptor antagonism with tezosentan was well tolerated with no new safety concerns emerging. These data further support the potential role of tezosentan in the treatment of patients with acute heart failure. PMID- 11502645 TI - Effect of hyperoxia on left ventricular function and filling pressures in patients with and without congestive heart failure. AB - STUDY OBJECTIVES: To determine the effects of hyperoxia on left ventricular (LV) function in humans with and without congestive heart failure (CHF). DESIGN: An acute physiologic study of the effect of hyperoxia on right-heart hemodynamics, LV contractility (peak positive rate of rise of LV pressure [+dP/dt]), time constant of isovolumic left ventricular relaxation (tau), and LV filling pressures. SETTING: Bayer Cardiovascular Clinical Research Laboratory at the Mount Sinai Hospital, Toronto, Ontario. PATIENTS: Sixteen patients with stable CHF and 12 subjects with normal LV function received the hyperoxia intervention. INTERVENTIONS: Patients received 21% O(2) by a nonrebreather mask, followed by 100% O(2) for 20 min, and 21% O(2) for a 10-min recovery period. RESULTS: In response to hyperoxia, there was a 22 +/- 6% increase in LV end-diastolic pressure (LVEDP) in the CHF group and a similar 29 +/- 14% increase in LVEDP in the normal LV function group (p < 0.05 for both; mean +/- SEM). Hyperoxia was also associated with a prolongation in tau of 10 +/- 2% in the CHF group (p < 0.05) and 8 +/- 2% in the normal LV function group (p < 0.05). No changes in +dP/dt were observed in either group. CONCLUSIONS: Hyperoxia was associated with impairment of cardiac relaxation and increased LV filling pressures in patients with and without CHF. These observations indicate that caution should be used in the administration of high inspired O(2) fractions to normoxic patients, especially in the setting of CHF. PMID- 11502646 TI - Assessment of cardiac stress from massive pulmonary embolism with 12-lead ECG. AB - BACKGROUND: Massive pulmonary embolism (PE) that causes severe pulmonary hypertension can produce specific ECG abnormalities. We hypothesized that an ECG scoring system would vary in proportion to the severity of pulmonary hypertension and would help to distinguish patients with massive PE from patients with smaller PE and those without PE. METHODS: A 21-point ECG scoring system was derived (relative weights in parentheses): sinus tachycardia (2), incomplete right bundle branch block (2), complete right bundle branch block (3), T-wave inversion in leads V(1) through V(4) (0 to 12), S wave in lead I (0), Q wave in lead III (1), inverted T in lead III (1), and entire S(1)Q(3)T(3) complex (2). ECGs obtained within 48 h prior to pulmonary arteriography were located for 60 patients (26 positive for PE, 34 negative for PE) and for 25 patients with fatal PE. RESULTS: Interobserver agreement (11 readers) for ECG score was good (Spearman r = 0.74). The ECG score showed significant positive relationship to systolic pulmonary arterial pressure (sPAP) in patients with PE (r = 0.387, p < 0.001), whereas no significant relationship was seen in patients without PE (r = - 0.08, p = 0.122). When patients were grouped by severity of pulmonary hypertension (low, moderate, severe), only patients with severe pulmonary hypertension from PE had a significantly higher ECG score (mean, 5.8 +/- 4.9). At a cutoff of 10 points, the ECG score was 23.5% (95% confidence interval [CI], 16 to 31%) sensitive and 97.7% (95% CI, 96 to 99%) specific for the recognition of severe pulmonary hypertension (sPAP > 50 mm Hg) secondary to PE. In 25 patients with fatal PE, the ECG score was 9.5 +/- 5.2. CONCLUSIONS: The derived ECG score increases with severity of pulmonary hypertension from PE, and a score > or = 10 is highly suggestive of severe pulmonary hypertension from PE. PMID- 11502647 TI - Can peak expiratory flow measurements estimate small airway function in asthmatic children? AB - BACKGROUND: Asthma is characterized in part by small airways dysfunction. Peak expiratory flow (PEF) measurement has been suggested by all international guidelines as an important tool in asthma management. The correlation between PEF and FEV(1) but not with forced expired flow at 50% of vital capacity (FEF(50)) is well-established. STUDY OBJECTIVE: To determine the value of PEF measurement as a predictor of small airways status as expressed by FEF(50). DESIGN: Analysis of the association between PEF and FEF(50) in single and multiple determinations. PATIENTS: One hundred eleven asthmatic children (mean age, 11.8 years), grouped in the following way according to FEV(1) values: within normal range (n = 46); mildly reduced FEV(1) (n = 44); and moderately/severely reduced FEV(1) (n = 21). RESULTS: Overall, FEF(50) and PEF were significantly correlated (r = 0.49; p < 0.0001). However, in 41.6% of the patients, the actual FEF(50) differed by > 20% from the calculated FEF(50). PEF has a high specificity (82.4%) but a poor sensitivity (51.7%) to detect FEF(50) status. PEF was better able to reflect abnormal FEF(50) in the patients with more severe asthma and to reflect normal FEF(50) values in the healthier patients. In patients with multiple measurements (n = 40), the correlation between FEF(50) and PEF was significantly better than that derived from a single determination (multiple measurements r = 0.77; single measurement, r = 0.49). CONCLUSIONS: Although PEF is an important tool in the management of asthmatic patients, it does not yield a complete picture because it is not sensitive in detecting small airways function. It is best used at home along with regular spirometry measurements at the clinic. PEF may serve as a better index of changes in small airways function once an individual regression is determined. PMID- 11502648 TI - Spirometry reference values for American Indian adults: results from the Strong Heart Study. AB - STUDY OBJECTIVE: To derive spirometry normative values from a large population of American Indian adults and compare them to reference values for white adults. DESIGN: Pulmonary function was assessed using spirometry in participants of the Strong Heart Study, a multicenter, community-based, prospective study of cardiovascular risk factors and disease in American Indians, utilizing American Thoracic Society guidelines and a vigorous quality assurance program. SETTING: Central Arizona, southwestern Oklahoma, central South Dakota, and northeastern North Dakota. PARTICIPANTS: Acceptable spirometry results were obtained from 1,619 women and 1,005 men aged 45 to 74 years. RESULTS: Internal reference values and normal ranges for FEV(1), FVC, and the FEV(1)/FVC ratio were derived from a healthy subgroup of 253 women and 190 men, identified by excluding participants with factors associated with a lower FEV(1). Ten percent of the entire cohort (269 of 2,624 subjects) had airways obstruction, as defined by an FEV(1)/FVC below the lower limit of the normal (LLN) using the internal reference equations. After allowing for measurement "noise," 31 participants were below the LLN using reference equations for white adults from the large National Health and Nutrition Examination Study (NHANES) III study but were normal using the internal reference equations (1.3% false-positive), while 27 participants were classified as normal using NHANES III equations but had airways obstruction using the internal reference equations (1.2% false-negative). Similarly low misclassification rates were seen for a low FVC (prevalence, 17.6%). CONCLUSION: For clinical purposes, NHANES III spirometry reference equations for white adults may be used when testing American Indian women and men aged 45 to 74 years. PMID- 11502649 TI - Exhaled carbon monoxide and nitric oxide in COPD. AB - STUDY OBJECTIVES: To investigate whether exhaled carbon monoxide (CO) and nitric oxide (NO) could be used as noninvasive in vivo biomarkers of oxidative stress in the lungs of patients with COPD. DESIGN: Single-center cross-sectional study. PATIENTS: Ten healthy nonsmokers, 12 smokers, 15 stable ex-smokers with COPD, and 15 stable current smokers with COPD. INTERVENTIONS: Subjects attended the outpatient clinic on one occasion for pulmonary function tests and exhaled CO and NO measurements. MEASUREMENTS AND RESULTS: Mean (+/- SEM) CO levels in ex-smokers with COPD were higher (7.4 +/- 1.9 ppm; p < 0.05) than in nonsmoking control subjects (3.0 +/- 0.3 ppm) but were lower than in current smokers with COPD (20.0 +/- 2.6 ppm; p < 0.001). There was no correlation between exhaled CO and NO. There was no correlation between CO and lung function tests in any group of patients. Exhaled NO was higher in ex-smokers with COPD (12.0 +/- 1.0 parts per billion [ppb]; p < 0.001) than in healthy nonsmokers (6.5 +/- 0.6 ppb) and in current smokers with COPD (7.6 +/- 1.1 ppb; p < 0.01) compared to healthy smokers (3.3 +/- 0.4 ppb). Ex-smokers with COPD had higher exhaled NO levels than did current smokers with COPD (p < 0.001) There was a negative correlation between exhaled NO and FEV(1) in both ex-smokers with COPD (r = -0.60; p < 0.02) and current smokers with COPD (r = -0.59; p < 0.02). CONCLUSION: The measurement of exhaled CO and NO may represent a new method for the noninvasive monitoring of airway inflammation and oxidant stress in COPD ex-smokers. Exhaled CO and NO are strongly affected by cigarette smoking, which limits their usefulness as biomarkers in current smokers. PMID- 11502650 TI - Evaluation of right ventricular performance with a right ventricular ejection fraction thermodilution catheter and MRI in patients with pulmonary hypertension. AB - STUDY OBJECTIVES: We sought to compare catheter studies using a right ventricular ejection fraction (REF) catheter together with echocardiography and MRI in patients with pulmonary hypertension. PATIENTS AND METHODS: We compared hemodynamic findings, echocardiography, and MRI studies in 16 patients with pulmonary hypertension. Six healthy volunteers served as control subjects for the MRI studies. RESULTS: MRI imaging provided accurate assessment of cardiac output in all but two patients. As compared with MRI, the REF catheter constantly underestimated the REF and overestimated right ventricular volumes in patients with pulmonary hypertension. REF, end-systolic and end-diastolic right ventricular volumes, and right ventricular muscle mass, as determined by MRI, were almost identical in patients with preserved cardiac function and those with low-output failure. The only factor that was different in both groups was the severity of tricuspid regurgitation. CONCLUSION: Right ventricular dimensions and muscle mass do not differ in patients with pulmonary hypertension who have low cardiac output and those who do not. According to our results, the major determinant of cardiac output in these patients appears to be the severity of tricuspid regurgitation. The REF catheter provides invalid data on right ventricular dimensions in patients with pulmonary hypertension. PMID- 11502651 TI - Pulmonary complications in cardiac transplant recipients. AB - BACKGROUND: The incidence of pulmonary complications in heart transplant recipients has not been extensively studied. We report pulmonary complications in 159 consecutive adult orthotopic heart transplantations (OHTs) performed in 157 patients. MATERIALS AND METHODS: Retrospective review of medical records. RESULTS: Forty-seven of 159 recipients (29.9%) had 81 pulmonary complications. Pneumonia was the most common (n = 27), followed by bronchitis (n = 15), pleural effusion (n = 10), pneumothorax (n = 7), prolonged respiratory failure requiring tracheotomy (n = 7), and obstructive sleep apnea syndrome (n = 6). All patients with late-onset (> 6 months after transplantation) community-acquired bacterial pneumonia presented with fever, cough, and a new lobar consolidation on the chest radiograph, and responded promptly to empiric antibiotics without undergoing an invasive diagnostic procedure. In contrast, early-onset nosocomial bacterial pneumonias carried a 33.3% mortality rate. A positive tuberculin skin test result was associated with a significantly higher rate of pulmonary complications (62.5% vs 26.8%, p = 0.007). Lung cancer and posttransplant lymphoproliferative disorder (PTLD) developed exclusively in 6 of the 61 patients (8.1%) who received induction immunosuppression with murine monoclonal antibody (OKT3). CONCLUSION: Pulmonary complications are common following heart transplantation, occurring in 29.9% of recipients, and are attributed to pneumonia of primarily bacterial origin in one half of cases. Late-onset community-acquired pneumonia carried an excellent prognosis following empiric antibiotic therapy, suggesting that in the appropriate clinical setting invasive diagnostic procedures are unnecessary. Analogous to reports in other solid-organ transplant recipients, induction therapy with OKT3 was associated with an increased incidence of lung cancer and PTLD. Overall, the development of pulmonary complications after OHT has prognostic significance given the higher mortality in this subset of patients. PMID- 11502652 TI - Clinicoradiologic features of pleuropulmonary Paragonimus westermani on Kyusyu Island, Japan. AB - STUDY OBJECTIVE: Recently, the number of new cases of Paragonimus westermani in humans has gradually increased, and paragonimiasis is a re-emerging public health issue in Kyusyu, Japan. We review our recent experience with pleuropulmonary Paragonimus westermani. PATIENTS: Pulmonary paragonimiasis was diagnosed in 13 patients at the Third Department of Internal Medicine, Miyazaki Medical College between 1993 and 1999. RESULTS: Both sputum and bronchoscopic examinations revealed ova in four of nine patients; bronchoscopy yielded ova in two additional patients. Twelve patients (92%) had respiratory symptoms, including cough (92%), sputum and/or hemoptysis (92%), and chest pain (46%). Chest radiography and CT showed pleural lesions (62%) and parenchymal lesions (92%). Of note was the high frequency of solitary nodular lesions (62%), mimicking lung cancer, tuberculosis, or fungal diseases. Immunodiagnosis and bronchoscopic examination were also useful for diagnosis. Praziquantel treatment was very effective and had minimal side effects. One patient required surgical decortication for empyema in spite of treatment with praziquantel. Eosinophilia was noted in peripheral blood and body fluids, which was probably due to increased levels of interleukin-5. CONCLUSIONS: Our findings indicate that our patients with Paragonimus westermani presented with a wide variety of radiographic findings, which were different from the classic presentations reported earlier. Bronchoscopic examination and serologic tests are very useful for accurate diagnosis. As dietary habits change and international transportation increases, it appears likely that paragonimiasis will also increase in frequency in various parts of the world. PMID- 11502653 TI - Mediastinal lymph node sampling following positron emission tomography with fluorodeoxyglucose imaging in lung cancer staging. AB - OBJECTIVES: To evaluate the predictive accuracy as well as the rates of false positive and false-negative results of CT and positron emission tomography (PET) fluorodeoxyglucose (FDG) imaging in detecting the metastatic intrathoracic lymph nodes in patients with suspected or proven non-small cell lung cancer (NSCLC). Our other objective was to determine the need for routine invasive sampling procedure in confirming PET/CT staging results. METHODS: The results of CT and PET-FDG scanning in 77 patients with suspected or proven NSCLC were correlated with the histologic findings of hilar/mediastinal lymph node sampling using mediastinoscopy, open biopsy, thoracotomy, or thoracotomy with resection. Patients were then classified into resectable and unresectable groups based initially on PET results and compared to histologic findings. RESULTS: The sensitivity, specificity, and accuracy of CT and PET for detecting metastatic lymphadenopathy were 68%, 61%, 63%, and 87%, 91%, and 82%, respectively. A change of management with routine sampling following PET was seen in five of six patients (83%) with false-positive findings (13%) but in none of four patients (9%) with false-negative findings. CONCLUSION: The false-positive findings of PET FDG imaging affected selection of treatment in 83% of patients. However, false negative results did not change management in any patient. This could potentially prevent unnecessary invasive thoracotomy, mediastinoscopy, or other sampling procedures in patients with negative PET results. PMID- 11502654 TI - Outcome prediction of emergency patients by noninvasive hemodynamic monitoring. AB - OBJECTIVES: We used noninvasive hemodynamic monitoring in the initial resuscitation beginning in the emergency department (ED) for the following reasons: (1) to describe early survivor and nonsurvivor patterns of emergency patients in terms of cardiac, pulmonary, and tissue perfusion deficiencies; (2) to measure quantitatively the net cumulative amount of deficit or excess of the monitored functions that correlate with survival or death; and (3) to explore the use of discriminant analysis to predict outcome and evaluate the biological significance of monitored deficits. METHODS: This is a descriptive study of the feasibility of noninvasive monitoring of patients with acute emergency conditions in the ED to evaluate and quantify hemodynamic deficits as early as possible. The noninvasive monitoring systems consisted of a bioimpedance method for estimating cardiac output together with pulse oximetry to reflect pulmonary function, transcutaneous oxygen tension to reflect tissue perfusion, and BP to reflect the overall circulatory status. These continuously monitored noninvasive measurements were used to prospectively evaluate circulatory patterns in 151 consecutively monitored severely injured patients beginning with admission to the ED in a university-run county hospital. The net cumulative deficit or excess of each monitored parameter was calculated as the cumulative difference from the normal value vs the time-integrated monitored curve for each patient. The deficits of cardiac, pulmonary, and tissue perfusion functions were analyzed in relation to outcome by discriminant analysis and were cross-validated. RESULTS: The mean (+/- SEM) net cumulative excesses (+) or deficits (-) from normal in surviving vs nonsurviving patients, respectively, were as follows: cardiac index (CI), +81 +/- 52 vs -232 +/- 138 L/m(2) (p = 0.037); arterial hemoglobin saturation, -1 +/- 0.3 vs -8 +/- 2.6%/h (p = 0.006); and tissue perfusion, +313 +/- 88 vs -793 +/- 175, mm Hg/h (p = 0.001). The cumulative mean arterial BP deficit for survivors was 10 +/- 13 mm Hg/h, and for nonsurvivors it was -57 +/- 24 mm Hg/h (p = 0.078). CONCLUSIONS: Noninvasive monitoring systems provided continuously monitored on line displays of data in the early postadmission period from the ED to the operating room and to the ICU for early recognition of circulatory dysfunction in short-term emergency conditions. Survival was predicted by discriminant analysis models based on the quantitative assessment of the net cumulative deficits of CI, arterial hypoxemia, and tissue perfusion, which were significantly greater in the nonsurvivors. PMID- 11502655 TI - Factors associated with reintubation in intensive care: an analysis of causes and outcomes. AB - INTRODUCTION: Reports on reintubation have focused on patients in whom planned extubation has been unsuccessful or those who have been accidentally extubated. However, reintubation is often required in events not related to accidental extubation. These cases have not been well described previously. OBJECTIVES: To examine the causes, outcomes, and contributing factors associated with patients who required reintubation for events not including accidental extubation. METHODS: Appropriate reintubation incidents were extracted from the Australian Incident Monitoring Study in Intensive Care database and analyzed using descriptive methodology. RESULTS: One hundred forty-three incidents were identified with prominent precipitating events, including tube malposition (17%), securing/taping problems (17%), pilot tube/cuff problem (16%), blocked/kinked airway (14%), failed extubation (14%), and poor planning for extubation (6%). Narrative description of morbidity included hypoxia in 25% of reports, hypercarbic respiratory failure in 12%, aspiration in 7%, sputum retention in 7%, and cardiac arrhythmias in 6%. The reporter selected "major physiologic complications" and "prolonged hospital stay" as prominent adverse outcomes in 52% and 16% of patients, respectively. Major factors contributing to reintubation involved "error of judgement/problem recognition" (identified in 62% of reports), "high unit activity" (20%), "difficult patient habitus" (26%), and "lack of patient cooperation" (14%). Rechecking patient and equipment, and skilled assistance were prominent factors in limiting the adverse consequences of the incident. CONCLUSION: This study indicated that reintubation not related to accidental extubation resulted in major physiologic complications and potentially contributed to increased length of stay. Its findings suggest that the adequate provision of highly qualified, intensive-care-trained staff is essential for the avoidance or minimization of these incidents. PMID- 11502656 TI - The Macklin effect: a frequent etiology for pneumomediastinum in severe blunt chest trauma. AB - STUDY OBJECTIVES: To review the etiology and pathophysiology of pneumomediastinum in severe blunt trauma, with a special interest in one of its possible origins, the Macklin effect. The Macklin effect relates to a three-step pathophysiologic process: blunt traumatic alveolar ruptures, air dissection along bronchovascular sheaths, and spreading of this blunt pulmonary interstitial emphysema into the mediastinum. The clinical relevance of the Macklin effect was also evaluated. SETTING: A university hospital serving as a reference trauma center. PATIENTS: A selection of 51 patients with severe blunt trauma between 1995 and 2000. INCLUSION CRITERIA: Severe trauma or high-speed deceleration justifying chest CT; if chest CT demonstrated a pneumomediastinum, bronchoscopy and esophagoscopy were performed to rule out tracheobronchial or esophageal injury. DESIGN: Retrospective analysis of patients' clinical files, chest CT, and bronchoscopy and esophagoscopy reports. The Macklin effect was diagnosed when an air collection adjacent to a bronchus and a pulmonary vessel could be clearly identified on the chest CT. Clinical relevance of the Macklin effect was statistically evaluated regarding its repercussions on the pulmonary gas exchange function, the respective durations of intensive care and total hospital stay, and the associated injuries. RESULTS: Twenty (39%) Macklin effects and 5 tracheobronchial injuries (10%) were identified. One tracheobronchial injury occurred simultaneously with the Macklin effect. The presence of the Macklin effect affected neither the clinical profile nor the result of pulmonary gas analysis on hospital admission, but was associated with a significant (p < 0.001) lengthening of the intensive care stay. CONCLUSIONS: The Macklin effect is present in 39% of severe blunt traumatic pneumomediastinum detected by CT. Its identification does not rule out a tracheobronchial injury. The Macklin effect reflects severe trauma, since it is associated with significantly prolonged intensive care stay. PMID- 11502657 TI - Hantavirus pulmonary syndrome due to Andes virus in Temuco, Chile: clinical experience with 16 adults. AB - STUDY OBJECTIVES: To describe the clinical features and laboratory abnormalities of 16 adults with confirmed Hantavirus pulmonary syndrome (HPS) due to Andes virus in Temuco, Chile. DESIGN: A retrospective chart review abstracting clinical, radiologic, laboratory, and epidemiologic data. SETTING: ICU of the university teaching hospital in Temuco, Chile. PATIENTS: Sixteen patients with HPS treated between 1997 and 1999. RESULTS: Patients were aged from 19 to 45 years, 82% were men, and 88% were farm or timber workers with occupational acquisition of HPS. After an incubation period ranging from 5 to 25 days, a prodromal influenza-like phase frequently was accompanied by abdominal symptoms. From 1 to 7 days later, respiratory insufficiency and hemodynamic instability suddenly appeared. In 81%, hemorrhage was evident; in 63%, moderate-to-severe bleeding occurred. The most prominent laboratory abnormalities were hemoconcentration, leukocytosis, thrombocytopenia, altered partial thromboplastin time (PTT), creatine kinase, transaminases, and hyponatremia. Creatinine elevation was common, with clinical importance in two patients. All patients had severe hypoxemia and pulmonary edema. Fifteen patients received supportive treatment, and 5 patients were treated with corticosteroids. The mortality rate was 43.8%. CONCLUSIONS: Bad prognostic factors appeared to be severe hypotension, lower PaO(2)/fraction of inspired oxygen values, prolonged PTT, hemorrhage, greater volume load, and profuse bronchorrhea. The effects of treatment with corticosteroids could not be determined. Hemorrhage and renal involvement were common in our patients, features not often described in the North American literature of Sin Nombre virus HPS. PMID- 11502658 TI - The occurrence of ventilator-associated pneumonia in a community hospital: risk factors and clinical outcomes. AB - STUDY OBJECTIVES: To prospectively identify the occurrence of ventilator associated pneumonia (VAP) in a community hospital, and to determine the risk factors for VAP and the influence of VAP on patient outcomes in a nonteaching institution. DESIGN: Prospective cohort study. SETTING: A medical ICU and a surgical ICU in a 500-bed private community nonteaching hospital: Missouri Baptist Hospital. PATIENTS: Between March 1998 and December 1999, all patients receiving mechanical ventilation who were admitted to the ICU setting were prospectively evaluated. INTERVENTION: Prospective patient surveillance and data collection. RESULTS: During a 22-month period, 3,171 patients were admitted to the medical and surgical ICUs. Eight hundred eighty patients (27.8%) received mechanical ventilation. VAP developed in 132 patients (15.0%) receiving mechanical ventilation. Three hundred one patients (34.2%) who received mechanical ventilation died during hospitalization. Logistic regression analysis demonstrated that tracheostomy (adjusted odds ratio [AOR], 6.71; 95% confidence interval [CI], 3.91 to 11.50; p < 0.001), multiple central venous line insertions (AOR, 4.20; 95% CI, 2.72 to 6.48; p < 0.001), reintubation (AOR, 2.88; 95% CI, 1.78 to 4.66; p < 0.001), and the use of antacids (AOR, 2.81; 95% CI, 1.19 to 6.64; p = 0.019) were independently associated with the development of VAP. The hospital mortality of patients with VAP was significantly greater than the mortality of patients without VAP (45.5% vs 32.2%, respectively; p = 0.004). The occurrence of bacteremia, compromised immune system, higher APACHE (acute physiology and chronic health evaluation) II scores, and older age were identified as independent predictors of hospital mortality. CONCLUSIONS: These data suggest that VAP is a common nosocomial infection in the community hospital setting. The risk factors for the development of VAP and risk factors for hospital mortality in a community hospital are similar to those identified from university-affiliated hospitals. These risk factors can potentially be employed to develop local strategies for the prevention of VAP. CLINICAL IMPLICATIONS: ICU clinicians should be aware of the risk factors associated with the development of VAP and the impact of VAP on clinical outcomes. More importantly, they should cooperate in the development of local multidisciplinary strategies aimed at the prevention of VAP and other nosocomial infections. PMID- 11502659 TI - Clinical predictors of respiratory failure and long-term outcome in black tar heroin-associated wound botulism. AB - STUDY OBJECTIVES: To our knowledge, the predictors of respiratory failure (RF) and long-term mechanical ventilation have not previously been examined in patients with wound botulism associated with black tar heroin use. DESIGN: Retrospective case series. SETTING: A large university hospital. PATIENTS: Twenty consecutive patients from 1991 to 1998 with a diagnosis of wound botulism associated with drug use as identified through chart records from a single institution. RESULTS: The predominant mode of drug use was subcutaneous (75%). Fifteen of 20 patients (75%) developed RF. The clinical presentation was similar in groups with RF and without RF. Of those patients who received antitoxin within 12 h of presentation, 57% required mechanical ventilation compared to 85% of patients receiving the antitoxin after 12 h. The median durations of mechanical ventilation were 11 days for those who received antitoxin within 12 h, and 54 days for those who did not receive antitoxin within 12 h. The duration of mechanical ventilation for patients receiving antibiotics within 12 h was 35 days vs 54 days for patients receiving antibiotics after 12 h. Early tracheostomy (< 10 days after initial intubation) was associated with a shorter duration of mechanical ventilation (median, 45 days vs 60 days, respectively). CONCLUSION: Early antitoxin administration may decrease the need for and duration of mechanical ventilation among patients with wound botulism. Early tracheostomy may be beneficial for patients with RF. PMID- 11502660 TI - Detection of viable myocardium by transvenous myocardial contrast echocardiography using harmonic power Doppler: canine model of acute coronary occlusion and reperfusion. AB - STUDY OBJECTIVE: To assess whether myocardial contrast echocardiography (MCE) using harmonic power Doppler (HPD) in conjunction with the transvenous contrast agent SHU 563A would be useful in detecting stunned but viable myocardium. DESIGN: Acute coronary occlusion (2 to 3 h) followed by 1 h of reperfusion was created in 10 dogs in an open-chest model. MEASUREMENTS AND RESULTS: Continuous harmonic B-mode for wall motion analysis and ECG triggered HPD for assessment of myocardial perfusion was employed during coronary occlusion and after reperfusion. Postmortem 2,3,5-triphenyltetrazolium chloride (TTC) staining was performed to verify infarction. Extent of wall motion abnormality (WMA), perfusion defect size, and anatomic infarct size (myocardial infarction [MI]) were analyzed in a 5-segment model. All 10 dogs showed WMA in 23 of 50 segments during coronary occlusion. In eight dogs, HPD detected perfusion defects in 18 of 50 segments. The concordance rate between WMA and perfusion defect was 86%. Mean linearized power (MLP) in segments with WMA was significantly lower compared to normal segments (60.7 +/- 38.9 vs 110.5 +/- 108.8, p < 0.05). After reperfusion, the extent of WMA was larger than the area of perfusion defect (percentage of left ventricular slice area): 30 +/- 13% vs 9 +/- 8%, p < 0.01. Eventual infarct size was 6 +/- 7%. WMAs were seen in 18 of 50 segments. TTC confirmed MI in 7 of 18 segments. MLP in segments with WMA but no MI was significantly higher compared to segments with WMA and MI (84.5 +/- 67.3 vs 13.2 +/- 9.6, p < 0.01). Thus, the extent of WMA after reperfusion was greater than the size of perfusion defect and eventual MI, indicating the presence of stunned but viable myocardium. CONCLUSION: MCE using HPD and the contrast agent SHU 563A can demonstrate the efficacy of reperfusion, identify necrotic regions, and aid in the recognition of stunned but viable myocardium. This approach could be useful clinically in patients with acute MI undergoing reperfusion therapy. PMID- 11502661 TI - An animal model of response and nonresponse to inhaled nitric oxide in endotoxin induced lung injury. AB - STUDY OBJECTIVE: Oxygenation may be improved in 40 to 60% of ARDS patients by inhalation of nitric oxide (NO). We have studied the response to inhaled NO in porcine acute lung injury 4 h and 6 h after onset of a 2-h endotoxin infusion (30 microg/kg/h), hypothesizing that a responder may change to a nonresponder over time and with progression of lung injury. DESIGN: Animal study. SETTING: Experimental laboratory in a university hospital. INTERVENTIONS AND MEASUREMENTS: We studied eight pigs under general anesthesia (mean weight, 26.2 kg) receiving mechanical ventilation adjusted to normocapnia, with a fraction of inspired oxygen (FIO(2)) of 0.5 to 1.0. Blood gases, endotoxin concentration, and central hemodynamics were measured hourly, and ventilation-perfusion (/) relationships were assessed by multiple inert gas elimination technique before and after inhalation of NO. NO was delivered at 40 ppm for 10 min at 4 h and 6 h of endotoxin exposure. RESULTS: Seven of eight pigs were responders to NO at 4 h, defined as a > or = 20% increase in oxygenation index (PaO(2)/FIO(2)) [223 +/- 43 to 330 +/- 56 mm Hg; p = 0.001]. The same pigs exhibited a > or = 20% fall in mean pulmonary artery pressure (39.4 +/- 2.2 to 30.0 +/- 2.1 mm Hg; p < 0.001). The response correlated to the perfusion to "normal /" regions (r = - 0.82) and negatively to shunt and dead space ventilation (r = 0.76 and r = 0.87, respectively). At 6 h, seven of eight pigs were nonresponders, despite unaltered hemodynamics and gas exchange. Correlations at 4 h between physiologic variables and response to NO were abolished. The logarithmic SDs of the perfusion distribution, a measure of the degree of / mismatch, increased significantly from 4 to 6 h (p = 0.04). CONCLUSION: Response to inhaled NO is abolished over time in endotoxin-induced ARDS pig lungs. The response seems to be related to the degree of / mismatch, which may indicate an important role of hypoxic pulmonary vasoconstriction. PMID- 11502662 TI - The effect of helium on ventilator performance: study of five ventilators and a bedside Pitot tube spirometer. AB - OBJECTIVE: To assess in vitro the performance of five mechanical ventilators Siemens 300 and 900C (Siemens-Elma; Solna, Sweden), Puritan Bennett 7200 (Nellcor Puritan Bennett; Pleasanton, CA), Evita 4 (Dragerwerk; Lubeck, Germany), and Bear 1000 (Bear Medical Systems; Riverside CA)-and a bedside sidestream spirometer (Datex CS3 Respiratory Module; Datex-Ohmeda; Helsinki, Finland) during ventilation with helium-oxygen mixtures. DESIGN: In vitro study. SETTING: ICUs of two university-affiliated hospitals. METHODS AND MEASUREMENTS: Each ventilator was connected to 100% helium through compressed air inlets and then tested at three to six different tidal volume (VT) settings using various helium-oxygen concentrations (fraction of inspired oxygen [FIO(2)] of 0.2 to 1.0). FIO(2) and VT were measured with the Datex CS3 spirometer, and VT was validated with a water displacement spirometer. MAIN RESULTS: The Puritan Bennett 7200 ventilator did not function with helium. With the other four ventilators, delivered FIO(2) was lower than the set FIO(2). For the Siemens 300 and 900C ventilators, this difference could be explained by the lack of 21% oxygen when helium was connected to the air supply port, while for the other two ventilators, a nonlinear relation was found. The VT of the Siemens 300 ventilator was independent of helium concentration, while for the other three ventilators, delivered VT was greater than the set VT and was dependent on helium concentration. During ventilation with 80% helium and 20% oxygen, VT increased to 125% of set VT for the Siemens 900C ventilator, and more than doubled for the Evita 4 and Bear 1000 ventilators. Under the same conditions, the Datex CS3 spirometer underestimated the delivered VT by about 33%. CONCLUSIONS: At present, no mechanical ventilator is calibrated for use with helium. This investigation offers correction factors for four ventilators for ventilation with helium. PMID- 11502663 TI - Association of p53 gene mutation and telomerase activity in resectable non-small cell lung cancer. AB - PURPOSE: Mutation of the p53 gene and deregulation of telomerase may be essential for canceration in some malignant diseases. However, relationships between these occurrences have not yet been clarified. We examined the roles of p53 gene mutation and telomerase activity relative to the clinical and pathologic features of non-small cell lung carcinoma (NSCLC). METHODS: Frozen sections of 40 surgically resected NSCLC specimens were used. DNA extracted from fresh tumor specimens was analyzed with polymerase chain reaction (PCR), single-strand conformation polymorphism (SSCP) method, to screen alterations in the p53 gene. Exons showing aberrant band shifts on SSCP were reamplified, and the PCR products were directly sequenced. In addition, the telomerase activity of the same specimens was analyzed quantitatively with the fluorescence-based telomeric repeat amplification protocol assay, and the total product generated (TPG) method. Clinical and pathologic parameters were evaluated using a statistical analysis system. RESULTS: Mutations of the p53 gene relevant to an altered protein were confirmed in 19 of 40 specimens (47.5%). The TPG of 40 specimens was 75.24 +/- 15.55 (mean +/- SE). The TPG of the 19 specimens positive for p53 gene mutation was significantly higher than that of the 21 specimens negative for p53 gene mutation. Furthermore, the degree of cell differentiation was significantly correlated with both p53 gene mutation and high telomerase activity. CONCLUSIONS: p53 gene mutation and high telomerase activity cooperate to induce tumorigenesis and low-grade differentiation in NSCLC. Simultaneous occurrence of p53 gene mutation and high telomerase activity may be relevant to the grade of malignancy in NSCLC. PMID- 11502664 TI - Increased expression of the chemoattractant cytokines eotaxin, monocyte chemotactic protein-4, and interleukin-16 in induced sputum in asthmatic patients. AB - BACKGROUND: Induced sputum from asthmatic patients has been recently used to assess inflammatory cells. We have previously reported an increased expression of Th-2-type cytokines in induced sputum of asthmatic patients. C-C chemokines, particularly eotaxin and monocyte chemotactic protein (MCP)-4, are associated with eosinophilic infiltration. Interleukin (IL)-16 is associated with chemotactic activity for CD4+ cells. Chemokine expression in BAL and bronchial biopsy specimens has been demonstrated in asthmatic airways, but not in induced sputum. METHODS: We examined whether eotaxin, MCP-4, and IL-16 expression could be detected in induced sputum of asthmatic patients (n = 10), and whether the expression was increased compared to normal control subjects (n = 9). Eotaxin, MCP-4, and IL-16 immunoreactivity were determined by immunocytochemistry. In addition, inflammatory cells were investigated using markers for T cells (CD3), eosinophils (major basic protein [MBP]), macrophages (CD68), neutrophils (elastase), and epithelial cells (cytokeratin). RESULTS: Our results showed that there was a significant difference in the percentages of MBP-positive and epithelial cells between asthmatic patients and normal control subjects (p < 0.05). However, there was no difference between these two groups in the percentage of CD3-, elastase-, and CD68-positive cells. Immunoreactivity for eotaxin, MCP-4, and IL-16 was expressed in the induced sputum of all asthmatic patients, and expression of these chemotactic cytokines was significantly greater than in control subjects (p < 0.001, p < 0.005, and p < 0.001, respectively). CONCLUSIONS: This study showed that induced sputum could be used to detect chemokines in patients with bronchial asthma, and that the upregulation of chemotactic cytokines in the airways can be seen using noninvasive techniques. PMID- 11502665 TI - High concentrations of beta-chemokines in BAL fluid of patients with diffuse panbronchiolitis. AB - BACKGROUND: T cells are important cellular components of bronchial inflammation in diffuse panbronchiolitis (DPB). beta-Chemokines such as RANTES (regulated on activation, normal T-cell expressed and secreted) and macrophage inflammatory peptide (MIP)-1alpha are closely related to the migration of inflammatory cells into the lung. In this study, we investigate the contribution of beta-chemokines to the accumulation of T cells in the lungs of patients with DPB. PATIENTS AND METHODS: We determined the levels of beta-chemokines in BAL fluid (BALF) and the correlation between these levels and T-cell subsets in BALF of 23 patients with DPB and 16 healthy subjects by sandwich enzyme-linked immunosorbent assay and flow cytometry. RESULTS: Percentages of CD3+ human leukocyte antigen (HLA)-DR+, CD8+, and CD8+HLA-DR+ cells in BALF of patients were significantly higher than in the control BALF. The absolute number of CD8+HLA-DR+ cells was also higher in BALF of patients than in the control BALF (p < 0.0001). Phenotypic analysis of CD4+ cells in BALF showed a similar percentage of CD4+CD45RA+ cells and CD4+CD29+ cells in patients and normal subjects. The concentrations of RANTES and MIP 1alpha in BALF of patients with DPB were significantly higher than in BALF of normal subjects (p < 0.05). In addition, there was a significant correlation between the absolute number or percentage of CD8+HLA-DR+ cells and MIP-1alpha concentration in BALF. CONCLUSIONS: Our results suggest that the interaction between activated CD8+ T cells and MIP-1alpha may contribute to the pathogenesis of DPB. PMID- 11502666 TI - Plastic bronchitis and the role of bronchoscopy in the acute chest syndrome of sickle cell disease. AB - STUDY OBJECTIVES: To review the prevalence, clinical features, and role of bronchoscopy in patients with plastic bronchitis during the acute chest syndrome (ACS) of sickle cell disease (SCD). DESIGN: Eight-year review of clinical experience. SETTING: Tertiary referral children's hospital. PATIENTS: Twenty-six pediatric inpatients with 29 ACS episodes requiring diagnostic bronchoscopy. RESULTS: Of the pediatric inpatients with ACS who underwent bronchoscopy, plastic bronchitis was diagnosed in 21 of 29 episodes (72%). There was no difference in clinical features between the patients with and without plastic bronchitis. Bronchoscopy was an essential diagnostic tool, but its therapeutic benefits were doubtful. CONCLUSIONS: This is the first report of the prevalence of plastic bronchitis in patients with ACS of SCD. In our patient population, this condition was found to be common. The role of diagnostic bronchoscopy is essential. A large series, multicenter study is required to determine whether bronchoscopy and BAL are therapeutically beneficial when added to currently practiced supportive care. PMID- 11502667 TI - Pediatric fiberoptic bronchoscopy with a laryngeal mask airway. AB - BACKGROUND AND OBJECTIVES: Bedside flexible fiberoptic bronchoscopy (FFB) with sedation has been recognized as a diagnostic modality in children. In certain circumstances, general anesthesia with endotracheal intubation is advocated. This study evaluates the usefulness of the laryngeal mask airway (LMA) as an alternative to endotracheal intubation during pediatric FFB. DESIGN, SETTING, AND PATIENTS: Between July 1995 and June 2000, we studied 92 children (51 girls; age range, 1 through 15 years) in the operating theater of a major tertiary children's hospital. The LMA was used in children with atelectasis, diffuse infiltrates, and those who required BAL under general anesthesia. The size of the LMA was chosen to accommodate a bronchoscope appropriate for the child's weight and age. RESULTS: Procedures were well tolerated, no complications were observed, and oxygen saturation exceeded 95% in all patients. Major findings included mucoid impaction and purulent bronchial secretions, and BAL was successfully accomplished in all individuals. CONCLUSIONS: Diagnostic BAL or extraction of mucous plugs should be accomplished with optimal control of the airway under general anesthesia. The use of the LMA during FFB is safe, provides excellent patient comfort, and should be utilized as an alternative to endotracheal intubation. PMID- 11502668 TI - Bleomycin-induced pneumonitis. AB - The cytotoxic agent bleomycin is feared for its induction of sometimes fatal pulmonary toxicity, also known as bleomycin-induced pneumonitis (BIP). The central event in the development of BIP is endothelial damage of the lung vasculature due to bleomycin-induced cytokines and free radicals. Ultimately, BIP can progress in lung fibrosis. The diagnosis is established by a combination of clinical symptoms, radiographic alterations, and pulmonary function test results, while other disorders resembling BIP have to be excluded. Pulmonary function assessments most suitable for detecting BIP are those reflecting lung volumes. The widely used transfer capacity of the lungs for carbon monoxide appeared recently not to be specific when bleomycin is used in a polychemotherapeutic regimen. There are no proven effective treatments for BIP in humans, although corticosteroids are widely applied. When patients survive BIP, they almost always recover completely with normalization of radiographic and pulmonary function abnormalities. This review focuses on BIP, especially on the pathogenesis, risk factors, and its detection. PMID- 11502669 TI - Overnight pulse oximetry for sleep-disordered breathing in adults: a review. AB - Pulse oximetry is a well-established tool routinely used in many settings of modern medicine to determine a patient's arterial oxygen saturation and heart rate. The decreasing size of pulse oximeters over recent years has broadened their spectrum of use. For diagnosis and treatment of sleep-disordered breathing, overnight pulse oximetry helps determine the severity of disease and is used as an economical means to detect sleep apnea. In this article, we outline the clinical utility and economical benefit of overnight pulse oximetry in sleep and breathing disorders in adults and highlight the controversies regarding its limitations as presented in published studies. PMID- 11502670 TI - Mitochondrial disease: a pulmonary and critical-care medicine perspective. AB - The clinical spectrum of mitochondrial diseases has expanded dramatically in the last decade. Abnormalities of mitochondrial function are now thought to participate in a number of common adult diseases, ranging from exercise intolerance to aging. This review outlines the common presentations of mitochondrial disease in ICUs and in the outpatient setting and discusses current diagnostic and therapeutic options as they pertain to the pulmonary and critical care physician. PMID- 11502671 TI - The white chest. PMID- 11502673 TI - Video-assisted thoracoscopic surgery for catamenial hemoptysis. AB - Catamenial hemoptysis is a rare condition, and only 36 cases have been reported since the first published case. We describe a woman with catamenial hemoptysis recurring over 8 years. The lesion was diagnosed using chest CT scan during menses and was also visualized clearly via thoracoscopy. The patient was treated successfully with a partial resection of the lung using video-assisted thoracic surgery (VATS) and has been asymptomatic for 14 months since the operation. We suggest that VATS for catamenial hemoptysis is a more effective treatment than medical therapy. PMID- 11502674 TI - Weakness, daytime somnolence, cough, and respiratory distress in a 77-year-old man with a history of childhood polio. PMID- 11502675 TI - The role of passive immunization in hiv-positive patients : a case report. AB - An HIV-positive patient presented with pulmonary tuberculosis as her AIDS defining diagnosis in 1993 and was effectively treated with 12 months of standard antituberculosis medications (isoniazide, rifampin, and pyrazinamide for 2 months). She received zidovudine for 6 weeks at the time of her diagnosis; however, because of patient preference, she has not received subsequent standard HIV medications (7 years). Her CD4 count at the time of diagnosis (1993) was 297/microL. Monthly passive immunotherapy was administered (fresh frozen plasma from HIV-negative blood donors with a significant titer for the anti-vasoactive intestinal peptide [VIP]/NTM antibody) from December 1993 to June 1994. Her CD4 count increased to > 400/microL during the passive immunotherapy and has remained stable for the past 6 years. The rational for the use of anti-VIP/NTM antibodies preparations in HIV, the possible mode of action of anti-VIP/NTM antibodies, the use of Ig preparations, and the role of exercise as a natural source of anti VIP/NTM antibodies are discussed. This case report supports the potential therapeutic use of anti-VIP antibodies for treatment of HIV disease. PMID- 11502676 TI - Pneumocystis carinii pneumonia in pregnancy. AB - OBJECTIVE: To report five new cases of Pneumocystis carinii pneumonia (PCP) and to review and analyze the existing reports on the subject. METHOD: Five new cases of PCP during pregnancy are described. The cases, case series, and related articles on the subject in the English language were identified through a comprehensive MEDLINE search and reviewed. RESULTS: More than 80% of women with AIDS are of reproductive age, and PCP is the most common cause of AIDS-related death in pregnant women in the United States. Among 22 reviewed cases, the mortality rate was 50% (11 of 22 patients), which is higher than that usually reported for HIV-infected individuals with PCP. Respiratory failure developed in 13 patients (59%), and mechanical ventilation was therefore required, and the survival rate in patients requiring mechanical ventilation was 31%. Maternal and fetal outcomes were better in cases of PCP during the third trimester of the pregnancy. A variety of treatment regimens were used, including sulfamethoxazole trimethoprim (SXT) alone or in combination with pentamidine, steroids, and eflornithine. The survival rate in patients treated with SXT alone was 71% (5 of 7 patients) and for those treated with SXT and steroids was 60% (3 of 5 patients), with an overall survival rate in both groups of 66.6% (8 of 12 patients). CONCLUSION: PCP has a more aggressive course during pregnancy, with increased morbidity and mortality. Maternal and fetal outcomes remain dismal. Treatment with SXT, compared to other therapies, may result in an improved outcome. Withholding appropriate PCP prophylaxis may adversely affect maternal and fetal outcomes. PMID- 11502677 TI - Acute myopericarditis after diphtheria, tetanus, and polio vaccination. AB - We report the first case of myopericarditis after triple vaccination against diphtheria, tetanus, and poliovirus in a young adult. He presented with fever, acute chest pain, and diffuse ST-segment elevation 2 days after vaccination. Two dimensional echocardiography findings were normal. Endomyocardial biopsy showed interstitial edema with diapedesis of erythrocytes. Laboratory findings showed inflammatory syndrome and elevated circulating immune complexes. He recovered within a few days with high-dose aspirin treatment and was without complications at 3-month follow-up. We discuss the different hypotheses for infective or hypersensitivity myocarditis. PMID- 11502678 TI - Improved chest expansion in idiopathic scoliosis after intensive, multiple modality, nonsurgical treatment in an adult. AB - This case report documents a substantial increase in chest wall expansion in a middle-aged woman with stable right thoracic spinal curvature due to idiopathic scoliosis. Treatment involved intensive psychological and mobilization therapies, including comprehensive manipulative medicine treatments and daily manual traction. Over an 8-year period, a 6-cm increase in resting chest circumference (in the absence of weight gain) and a 7.5-cm increase in chest expansion were correlated with a substantial reduction of incidence of respiratory infections. PMID- 11502679 TI - TB or not TB: cavitary bronchiolitis obliterans organizing pneumonia mimicking pulmonary tuberculosis. AB - Two patients with subacute symptoms and signs compatible with pulmonary tuberculosis (TB) had right upper lobe cavitary infiltrates shown on chest radiography. In both patients, purified protein derivative and microbiologic testing excluded TB, and tissue examination yielded typical histologic changes of bronchiolitis obliterans organizing pneumonia (BOOP). Glucocorticoid therapy led to clinical and radiologic resolution. Though probably rare in this situation, BOOP should be considered in the differential diagnosis of patients presenting with clinical and radiologic features of pulmonary TB. PMID- 11502680 TI - Respiratory control during independent lung ventilation. AB - We describe the case of a lung transplant patient with primary graft failure and an emphysematous native lung, who displayed different respiratory rates between the transplanted lung and the native lung. Inflation of the native lung delayed the next inspiratory effort relative to inflation of the denervated transplanted lung. Synchronous inflation of both lungs required more pressure in each lung than when that lung was inflated with the contralateral lung near functional residual capacity, suggesting the two lungs compete for space within the thoracic cavity. PMID- 11502681 TI - Lysis of a left ventricular thrombus with recombinant tissue plasminogen activator. AB - A 23-year-old woman with peripartum cardiomyopathy presented with a 2.1 x 2.5-cm pedunculated, mobile, left ventricular thrombus and evidence of systemic embolization. Due to the patient's poor left ventricular function, thrombectomy was not a viable option. Treatment with high-dose IV heparin was initially utilized but was unsuccessful as the thrombus appeared to enlarge on echocardiography. An accelerated weight-adjusted dose of recombinant tissue plasminogen activator (rt-PA) successfully lysed the thrombus without evidence of embolization. Although rt-PA has been used for primary lysis of high-risk ventricular thrombi, this is the first documentation of successful lysis of a left ventricular thrombus in a patient with peripartum cardiomyopathy. PMID- 11502682 TI - Faulty design resulting in temporary pacemaker failure. AB - A 77-year-old man became asystolic 3 days after aortic valve replacement and coronary artery bypass surgery. A dual-chamber temporary pacemaker generator was turned on but failed to discharge; instead, an obscure error message appeared on the liquid crystal display of the pacemaker. The intensive-care nurses and physicians were unable to activate the pacemaker. We describe the pacemaker design that led to this instance of pacemaker failure. This case is important because it illustrates how a medical equipment design flaw can turn a human error into a potentially catastrophic event. PMID- 11502683 TI - How much incidence is enough? PMID- 11502684 TI - Surveillance bronchoscopy following insertion of silicone stents. PMID- 11502685 TI - Dr. Kahana's lesson: some more to learn. PMID- 11502686 TI - Augmentation therapy in alpha(1)-antitrypsin deficiency. PMID- 11502687 TI - Cardiac thrombus in Behcet disease. PMID- 11502688 TI - Ethics in decision making and patient autonomy. PMID- 11502689 TI - Metastatic lung cancer without regional lymph node swelling. PMID- 11502690 TI - Our antismoking campaign. PMID- 11502694 TI - Plasma nonesterified fatty acid concentration as a risk factor for sudden cardiac death: the Paris Prospective Study. PMID- 11502695 TI - Serum levels of the antiinflammatory cytokine interleukin-10 are decreased in patients with unstable angina. AB - BACKGROUND: Proinflammatory cytokines play a role in acute coronary events. However, the potential role of antiinflammatory cytokines in the modulation of the atherosclerotic process remains unknown. Interleukin (IL)-10, which is expressed in human atherosclerotic plaques, has potent deactivating properties in macrophages and T cells. The aim of this study was to assess whether serum concentrations of IL-10 differed between patients with unstable and stable angina pectoris. METHODS AND RESULTS: A total of 95 patients with angina pectoris and angiographically documented coronary artery disease were studied. Of these, 50 patients had chronic stable angina (with stable symptoms over 3 months), and 45 patients had Braunwald class IIIB unstable angina with ST-segment changes. Serum IL-10 and IL-6 concentrations were measured on admission using commercially available immunoassays. Serum IL-10 concentrations were lower in unstable angina patients compared with those who had chronic stable angina (28.4 versus 14.0 pg/mL; 95% CI, 9.8 to 19.0; P<0.0001), even after adjustment for variables that were significantly different on univariate analysis. IL-6 concentrations were higher in the unstable angina group (20.9 versus 11.4 pg/mL; 95% CI, 1.0 to 12.6; P=0.04). CONCLUSIONS: Patients with unstable angina had significantly lower serum IL-10 concentrations than did patients with chronic stable angina. This important finding is in keeping with previous data from animal model studies that suggest that IL-10 has a protective role in atherosclerosis. PMID- 11502696 TI - Functional evidence for a role of vascular chymase in the production of angiotensin II in isolated human arteries. AB - BACKGROUND: In human arteries, angiotensin-converting enzyme (ACE) inhibitors incompletely block the production of angiotensin (Ang) II from Ang I. This ACE independent production of Ang II appears to be caused by serine proteases, one of which presumably is chymase. However, several serine proteases may produce Ang II, and the exact role of chymase in the vascular production of Ang II has never been directly evaluated using selective chymase inhibitors. METHODS AND RESULTS: Rings of human mammary arteries were subjected to either Ang I or the chymase selective substrate [pro,(11) D-Ala(12)] Ang I in the absence or the presence of the ACE inhibitor captopril, the serine protease inhibitor chymostatin, or the selective chymase inhibitor C41. Captopril only partially inhibited (by 33%) the response to Ang I. In the absence of captopril, C41 markedly reduced (by 44%) the response to Ang I, and this effect was identical to that of chymostatin. C41 also significantly reduced the response to Ang I in the presence of captopril, although this inhibitory effect was slightly less than that of captopril in combination with chymostatin. [Pro,(11)D-Ala(12)] Ang I induced potent contractions that were not affected by captopril but were abolished by chymostatin and markedly reduced by C41. In addition, we found that prior treatment of the patients with an ACE inhibitor did not affect the in vitro response to Ang I (in the absence or the presence of captopril) or to [Pro,(11)D Ala(12)] Ang I. CONCLUSIONS: Our results reinforce the hypothesis that chymase is a major serine protease implicated in the ACE-independent production of Ang II in human arteries. PMID- 11502697 TI - Adenovirus-mediated gene transfer of VEGF(121) improves lower-extremity endothelial function and flow reserve. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) currently is being evaluated in clinical angiogenesis trials involving patients with peripheral arterial disease. We hypothesized that delivery of VEGF to the skeletal muscle of the lower extremity using an adenoviral vector (Ad(GV)VEGF(121.10)) would improve peripheral endothelial function. Accordingly, we investigated lower-extremity endothelial function in patients enrolled in a Phase I adenovirus-mediated gene delivery trial of VEGF(121.10). METHODS AND RESULTS: Blood flow to the index extremity was measured by thermodilution at baseline and 30 days after administration of Ad(GV)VEGF(121.10), in response to the infusion of endothelium dependent and -independent agonists (acetylcholine and nitroglycerin, respectively) into the ipsilateral femoral artery. There was no difference in basal flow before or after treatment with Ad(GV)VEGF(121.10). In response to acetylcholine (150 microg/min and 300 microg/min), there was a 0.9-fold (0.33+/ 0.03 to 0.32+/-0.03 L/min) and 1.2-fold (0.33+/-0.03 to 0.490+/-0.02 L/min) change in flow before Ad(GV)VEGF(121.10) treatment. After Ad(GV)VEGF(121.10) treatment, flow increased 2.4-fold (0.310+/-0.04 to 0.730+/-0.10 L/min) and 2.3 fold (0.31+/-0.04 to 0.7+/-0.08 L/min), respectively (P<0.05 before Ad(GV)VEGF(121.10) treatment versus after Ad(GV)VEGF(121.10) for both doses). Infusion of nitroglycerin resulted in a 1.8-fold increase in flow before Ad(GV)VEGF(121.10) (0.33+/-0.03 to 0.58+/-0.06 L/min) compared with a 2.4-fold increase (0.31+/-0.04 to 0.73+/-0.09 L/min) after Ad(GV)VEGF(121.10) (P=NS before Ad(GV)VEGF(121.10) versus after Ad(GV)VEGF(121.10)). Lower-extremity flow reserve increased in all patients in response to at least 1 dose of acetylcholine. Peak walking times increased concomitant with improvement in endothelial function. CONCLUSIONS: Adenoviral gene transfer of VEGF(121.10) appears to modulate endothelial function and lower-extremity flow reserve in patients with peripheral arterial disease. PMID- 11502698 TI - Circulating nonesterified fatty acid level as a predictive risk factor for sudden death in the population. AB - BACKGROUND: In ischemic conditions, concentration of circulating nonesterified fatty acids (NEFA) is increased and has a proarrhythmic effect that is responsible for ventricular tachyarrhythmias. In nonischemic patients, high NEFA plasma concentration has been shown to be associated with frequent premature ventricular complexes and increased familial risk of cardiovascular disease, but its relation to sudden death has not been studied. We assessed the role of circulating NEFA in sudden death in asymptomatic men in a long-term cohort study. METHODS AND RESULTS: A total of 5250 men employed by the city of Paris, aged 42 to 53 in 1967 to 1972, free of known ischemic cardiac disease, and included in the Paris Prospective Study I, completed a second annual examination and had fasting plasma circulating NEFA measured. Each subject underwent a physical examination and ECG, provided blood for laboratory tests, and answered questionnaires administered by trained interviewers. Vital status was obtained for each subject from specific inquiries until he retired; after retirement, it was obtained from death certificates. Body mass index, systolic and diastolic blood pressures, tobacco consumption, parental history of sudden death, fasting cholesterol level, and circulating NEFA concentration were independent factors associated with sudden death during follow up (average, 22 years). When adjusted for confounding factors, circulating NEFA concentration remained an independent risk factor for sudden death (relative risk, 1.70; 95% confidence interval, 1.21 to 2.13) but not for fatal myocardial infarction. CONCLUSIONS: Circulating NEFA concentration is an independent risk factor for sudden death in middle-aged men. Some form of primary prevention could be envisaged in subjects at high risk of sudden death. PMID- 11502699 TI - Decreased coagulability has no clinically relevant effect on atherogenesis: observations in individuals with a hereditary bleeding tendency. AB - BACKGROUND: Hemostasis affects ischemic cardiovascular disease through its role in formation of occluding arterial thrombi. Several studies suggest that hemostasis also might play a role in atherogenesis. We investigated whether individuals with an inherited bleeding tendency are protected against development of atherosclerosis. METHODS AND RESULTS: A total of 76 individuals with an inherited bleeding tendency (hemophilia and von Willebrand disease) and 142 healthy controls were included in the present study. Early atherosclerotic vessel wall changes were quantified by measurement of intima-media thickness in the carotid and femoral arteries by B-mode ultrasonography. To validate intima-media thickness measurements, measurements also were performed in 77 individuals with clinically proven atherosclerosis and in 34 healthy, age-matched controls. A large difference in intima-media thickness was found between individuals with proven atherosclerosis and healthy controls, in particular for the femoral artery (difference for carotid artery, 0.16 mm; femoral artery, 0.53 mm). Comparison between patients with a bleeding tendency and healthy controls showed only minimally reduced intima-media in femoral artery in individuals with a bleeding tendency (adjusted difference, -0.078 mm; 95% CI, -0.17 to 0.018 mm). Subgroup analysis revealed that in subjects with moderate to severe hemophilia, vessel walls were thinnest (adjusted difference, -0.10 mm; 95% CI, -0.27 to 0.061 mm). CONCLUSIONS: Hypocoagulability caused by hemophilia or von Willebrand disease has at most a limited effect on atherogenesis. PMID- 11502700 TI - Phased chest and abdominal compression-decompression versus conventional cardiopulmonary resuscitation in out-of-hospital cardiac arrest. AB - BACKGROUND: Several methods have been developed to improve the efficacy of mechanical resuscitation, because organ perfusion achieved with conventional manual resuscitation is often insufficient. In animal studies, phased chest and abdominal compression-decompression resuscitation by use of the Lifestick device has resulted in a better outcome compared with that of conventional resuscitation. In end-of-life patients, an increased coronary perfusion pressure was achieved. The aim of the present study was to determine the feasibility, safety, and efficacy of the Lifestick compared with conventional resuscitation in patients with sudden nontraumatic out-of-hospital cardiac arrest. METHODS AND RESULTS: The crews of 4 mobile intensive care units, staffed by an emergency physician and a paramedic, were trained to use the device. Fifty patients were randomized by sealed envelopes to either Lifestick (n=24) or conventional (n=26) resuscitation. No differences were found regarding demographic and logistical conditions between the groups. Nineteen of the patients (73%) with conventional resuscitation had ventricular fibrillation, 13 of whom survived to hospital admission (no survivals with other arrhythmias) and 7 were discharged. In contrast, in the Lifestick-CPR group, only 9 patients had ventricular fibrillation (38%; P=<0.02; OR, 2.5; 95% CI, 0.6 to 10.6). Four of these 9 patients and 5 of 15 patients with other arrhythmias survived to hospital admission, but none survived to hospital discharge. Autopsy in a subgroup of patients who died at the scene revealed less injuries with Lifestick than with conventional resuscitation. CONCLUSION: Lifestick resuscitation is feasible and safe and may be advantageous in patients with asystole or pulseless electric activity. PMID- 11502701 TI - Impact of smoking on clinical and angiographic restenosis after percutaneous coronary intervention: another smoker's paradox? AB - BACKGROUND: Recent studies have suggested that smokers may require less frequent repeated revascularization after percutaneous coronary intervention (PCI) compared with nonsmokers. However, the mechanism of this phenomenon is unknown. METHODS AND RESULTS: We examined the association between smoking and restenosis using pooled data from 8671 patients treated with PCI in 9 multicenter clinical trials. Clinical restenosis was examined in the cohort of 5682 patients who were assigned to clinical follow-up only. Angiographic restenosis was evaluated in the subset of 2989 patients who were assigned to mandatory angiographic restudy. Among those patients assigned to clinical follow-up only, target lesion revascularization (TLR) occurred in 6.6% of smokers and 10.1% of nonsmokers (P<0.001). After adjustment for baseline clinical and angiographic differences, the rate of TLR remained significantly lower in smokers with an adjusted relative risk of 0.69 (95% CI, 0.54 to 0.88). Among the angiographic cohort, there were no differences in the rates of angiographic restenosis or follow-up diameter stenosis in either univariate or multivariate analyses. This dissociation between clinical and angiographic restenosis was explained in part by reduced sensitivity to restenosis on the part of smokers and by the greater reluctance of smokers to seek medical attention despite recurrent angina. CONCLUSIONS: In patients undergoing contemporary PCI, cigarette smoking is associated with a lower rate of subsequent TLR without affecting angiographic restenosis. These findings have important implications for the follow-up of smokers after PCI and suggest that cross-study comparisons of rates of clinical restenosis must account for the potential confounding effect of smoking. PMID- 11502702 TI - Heart failure with a normal ejection fraction: is measurement of diastolic function necessary to make the diagnosis of diastolic heart failure? AB - BACKGROUND: The diagnosis of diastolic heart failure is generally made in patients who have the signs and symptoms of heart failure and a normal left ventricular (LV) ejection fraction. Whether the diagnosis also requires an objective measurement of parameters that reflect the diastolic properties of the ventricle has not been established. METHODS AND RESULTS: We hypothesized that the vast majority of patients with heart failure and a normal ejection fraction exhibit abnormal LV diastolic function. We tested this hypothesis by prospectively identifying 63 patients with a history of heart failure and an echocardiogram suggesting LV hypertrophy and a normal ejection fraction; we then assessed LV diastolic function during cardiac catheterization. All 63 patients had standard hemodynamic measurements; 47 underwent detailed micromanometer and echocardiographic-Doppler studies. The LV end-diastolic pressure was >16 mm Hg in 58 of the 63 patients; thus, 92% had elevated end-diastolic pressure (average, 24+/-8 mm Hg). The time constant of LV relaxation (average, 51+/-15 ms) was abnormal in 79% of the patients. The E/A ratio was abnormal in 48% of the patients. The E-wave deceleration time (average, 349+/-140 ms) was abnormal in 64% of the patients. One or more of the indexes of diastolic function were abnormal in every patient. CONCLUSIONS: Objective measurement of LV diastolic function serves to confirm rather than establish the diagnosis of diastolic heart failure. The diagnosis of diastolic heart failure can be made without the measurement of parameters that reflect LV diastolic function. PMID- 11502703 TI - Effect of aging on the prognostic significance of ambulatory systolic, diastolic, and pulse pressure in essential hypertension. AB - BACKGROUND: This study compared the relative prognostic significance of 24 hour intra-arterial ambulatory systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and pulse pressure (PP) parameters in middle aged versus elderly hypertensives. METHODS AND RESULTS: A total of 546 subjects aged <60 years and 142 subjects aged >/=60 years who had undergone baseline pretreatment 24-hour intra-arterial ambulatory blood pressure monitoring were followed for 9.2+/-4.1 years. Multivariate analysis showed that in younger subjects, 24-hour, daytime, and nighttime DBP, MAP, and SBP, when considered individually, were positively related to morbid events; DBP parameters provided the best predictive values. In the group >/=60 years (elderly group), 24-hour, daytime, and nighttime PP and SBP were the most predictive parameters, whereas ambulatory DBP and MAP measurements failed to provide any prognostic value. When 24-hour values of SBP and DBP were jointly included in the baseline model, DBP (z=2.02, P=0.04) but not SBP (z=-0.43, P=0.67) was related to outcome in younger subjects, whereas in the elderly group, SBP (z=3.33, P=0.001) was positively and DBP (z=-1.75, P=0.07) was negatively related to outcome. Clinic blood pressure measurements failed to provide any independent prognostic value in either age group. CONCLUSIONS: The relative prognostic significance of ambulatory blood pressure components depends on age; DBP parameters provided the best prognostic value in middle-aged individuals, whereas PP parameters were the most predictive in the elderly. This may reflect differing underlying hemodynamic mechanisms of hypertension in these age groups. PMID- 11502704 TI - Altered growth responses of pulmonary artery smooth muscle cells from patients with primary pulmonary hypertension to transforming growth factor-beta(1) and bone morphogenetic proteins. AB - BACKGROUND: Mutations in the type II receptor for bone morphogenetic protein (BMPR-II), a receptor member of the transforming growth factor-beta (TGF-beta) superfamily, underlie many cases of familial and sporadic primary pulmonary hypertension (PPH). We postulated that pulmonary artery smooth muscle cells (PASMCs) from patients with PPH might demonstrate abnormal growth responses to TGF-beta superfamily members. METHODS AND RESULTS: For studies of (3)H-thymidine incorporation or cell proliferation, PASMCs (passages 4 to 8) were derived from main pulmonary arteries. In control cells, 24-hour incubation with TGF-beta(1) (10 ng/mL) or bone morphogenetic protein (BMP)-2, -4, and -7 (100 ng/mL) inhibited basal and serum-stimulated (3)H-thymidine incorporation, and TGF beta(1) and BMPs inhibited the proliferation of serum-stimulated PASMCs. In contrast, TGF-beta(1) stimulated (3)H-thymidine incorporation (200%; P<0.001) and cell proliferation in PASMCs from PPH but not from patients with secondary pulmonary hypertension. In addition, BMPs failed to suppress DNA synthesis and proliferation in PASMCs from PPH patients. Reverse transcription-polymerase chain reaction of PASMC mRNA detected transcripts for type I (TGF-betaRI, Alk-1, ActRI, and BMPRIB) and type II (TGF-betaRII, BMPR-II, ActRII, ActRIIB) receptors. Receptor binding and cross-linking studies with (125)I-TGF-beta(1) confirmed that the abnormal responses in PPH cells were not due to differences in TGF-beta receptor binding. Mutation analysis of PASMC DNA failed to detect mutations in TGF-betaRII and Alk-1 but confirmed the presence of a mutation in BMPR-II in 1 of 5 PPH isolates. CONCLUSIONS: We conclude that PASMCs from patients with PPH exhibit abnormal growth responses to TGF-beta(1) and BMPs and that altered integration of TGF-beta superfamily growth signals may contribute to the pathogenesis of PPH. PMID- 11502705 TI - Shock reduction using antitachycardia pacing for spontaneous rapid ventricular tachycardia in patients with coronary artery disease. AB - BACKGROUND: Implantable cardioverter-defibrillators (ICDs) can terminate some ventricular tachycardias (VTs) painlessly with antitachycardia pacing (ATP). ATP has not routinely been applied for VT >188 bpm because of concerns about efficacy, risk of acceleration, and delay of definitive shock therapy. This prospective, multicenter study evaluated the efficacy of empirical ATP to terminate fast VT (FVT; >188 bpm). METHODS AND RESULTS: Two hundred twenty coronary artery disease patients received ICDs for standard indications. Empirical, standardized therapy was programmed so that all FVT episodes (average cycle length [CL] 240 to 320 ms, 250 to 188 bpm) were treated with 2 ATP sequences (8-pulse burst pacing train at 88% of the FVT CL) before shock delivery. A total of 1100 episodes of spontaneous ventricular tachyarrhythmias occurred during a mean of 6.9+/-3.6 months of follow-up. Fifty-seven percent were classified as slow VT (CL>/=320 ms), 40% as FVT (240 ms2-fold in obese Zucker rats compared with lean Zucker rats (0.21+/ 0.06 versus 0.08+/-0.03 mm(2), P<0.01). The neointimal area was markedly increased in the obese Zucker rats 7 days after injury (0.058+/-0.024 versus 0.033+/-0.009 mm(2), P<0.05) and persisted through 21 days. In both obese and lean Zucker rats, cell proliferation peaked in the media at 3 days (118.66+/ 84.28 versus 27.50+/-12.75 bromodeoxyuridine-labeled cells per cross section). In the intima, cell proliferation markedly increased beginning at day 3 and persisted through day 14 in the obese and lean Zucker rats (202.27+/-98.86 versus 35.71+/-20.54 bromodeoxyuridine-labeled cells at 7 days). CONCLUSIONS: The type II diabetic rat model, typifying insulin resistance, is associated with a propensity for neointima. The obese Zucker rat model may be an ideal diabetic model to further characterize the diabetic vascular response to injury. PMID- 11502709 TI - Effects of selective cyclooxygenase-2 inhibition on vascular responses and thrombosis in canine coronary arteries. AB - BACKGROUND: Prostanoid synthesis via the action of cyclooxygenase-2 (COX-2) is a component of the inflammatory response. Prostacyclin, a product of COX-2 in vascular endothelium, has important physiological roles, such as increasing blood flow to injured tissues, reducing leukocyte adherence, and inhibiting platelet aggregation. We examined the possibility that selective COX-2 inhibition could suppress the protective effects of prostacyclin, resulting in an alteration of the hemostatic balance and vascular tone. METHODS AND RESULTS: Circumflex coronary artery thrombosis was induced in dogs by vascular electrolytic injury. Orally administered celecoxib (COX-2 inhibition) or high-dose aspirin (HDA) (COX 1 and COX-2 inhibition) did not alter time to occlusive thrombus formation compared with controls (celecoxib 77.7+/-7.2 minutes, HDA 72.0+/-18.5 minutes, control 93.0+/-21.8 minutes). Oral HDA with an endothelial recovery period (HDA ER) (COX-1 inhibition) produced a significant increase in time to vessel occlusion (257.0+/-41.6 minutes). The observed increase in time to occlusion was abolished when celecoxib was administered to animals dosed with HDA-ER (80.7+/ 20.6 minutes). The vasomotor effect of endothelium-derived prostacyclin was examined by monitoring coronary flow during intracoronary administration of arachidonic acid or acetylcholine. In celecoxib-treated animals, vasodilation in response to arachidonic acid was reduced significantly compared with controls. CONCLUSIONS: The results indicate important physiological roles for COX-2-derived prostacyclin and raise concerns regarding an increased risk of acute vascular events in patients receiving COX-2 inhibitors. The risk may be increased in individuals with underlying inflammatory disorders, including coronary artery disease. PMID- 11502710 TI - Left ventricular remodeling in transgenic mice with cardiac restricted overexpression of tumor necrosis factor. AB - BACKGROUND: The mechanisms responsible for tumor necrosis factor (TNF)-induced LV structural remodeling in the adult heart are not known. METHODS AND RESULTS: We generated a line of transgenic mice (MHCsTNF) with cardiac restricted overexpression of TNF that develop progressive LV dilation/remodeling from 4 to 12 weeks of age. During the early phases of LV structural remodeling, there was a significant increase in total matrix metalloproteinase (MMP) activity that corresponded to a decrease in total myocardial fibrillar collagen content. As the MHCsTNF mice aged, there was a significant decrease in total MMP zymographic activity that was accompanied by an increase in total fibrillar collagen content. The changes in total MMP activity and myocardial fibrillar collagen content were related to a time- dependent increase in myocardial tissue inhibitor of metalloproteinases (TIMP)-1 levels, resulting in a significant time-dependent decrease in the MMP activity/TIMP level ratio in the MHCsTNF mice. To determine a possible mechanism for the increase in myocardial fibrosis, we also measured levels of TGF-beta(1) and TGF-beta(2) protein levels, which were shown to be significantly elevated in the hearts of the MHCsTNF mice. CONCLUSIONS: Our results suggest that progressive time-dependent changes in the balance between MMP activity and TIMP activity are responsible, at least in part, for the spectrum of TNF-induced changes in the myofibrillar collagen content that occur during LV structural remodeling in the MHCsTNF mice. PMID- 11502711 TI - His electrogram alternans reveal dual-wavefront inputs into and longitudinal dissociation within the bundle of His. AB - BACKGROUND: His electrogram (HE) amplitude and morphology changes were observed in our previous studies during transition from "fast" to "slow" atrioventricular nodal (AVN) conduction. This phenomenon and its significance for the dual-AVN electrophysiology are not well recognized and have not been studied. METHODS AND RESULTS: Experiments were performed on 17 healthy rabbit atrial-AVN preparations during standard programmed electrical pacing. HEs were mapped along the His bundle with roving surface electrodes, along with recording of cellular action potentials (APs). HEs recorded from the superior margin of the His bundle were of greater amplitude during basic beats and decreased substantially, by 42+/-19% (P<0.01), when premature A(1)A(2) shortened to 178+/-20 ms. In contrast, the HEs from the inferior margin increased dramatically, 2.9+/-1.7 times (P<0.01), during short A(1)A(2) and remained high until AVN block occurred. In addition, during long A(1)A(2), the superior HEs consistently preceded the inferior by 1.9+/-0.7 ms. In contrast, at short A(1)A(2), the superior HEs occurred 2.7+/-0.8 ms after the inferior. Cellular AP recordings demonstrated clearly the presence of and the transition between early (fast) and late (slow) excitation wavefronts that accompanied HE alternans. CONCLUSIONS: The morphological-electrophysiological evidence from the AV junction suggests that fast and slow wavefronts reach the His bundle differently, producing functional longitudinal dissociation into 2 domains. The characteristic HE alternans recorded from these domains are a new sensitive tool to determine the presence of distinctly different wavefronts and their participation in the conduction during reentrant or other arrhythmias. These findings provide further understanding of the mechanisms of dual-AVN electrophysiology. PMID- 11502713 TI - Learning from mistakes: the case of clinical electrophysiology: a perspective on evidence-based rhythmology. PMID- 11502712 TI - Plasminogen activator inhibitor-1 deficiency prevents hypertension and vascular fibrosis in response to long-term nitric oxide synthase inhibition. AB - BACKGROUND: Long-term inhibition of nitric oxide synthase (NOS) is known to induce hypertension and perivascular fibrosis. Recent evidence also suggests that long-term NOS inhibition induces expression of plasminogen activator inhibitor-1 (PAI-1) in vascular tissues and that PAI-1 may contribute to the development of fibrosis after chemical or ionizing injury. On the basis of these observations, we hypothesized that PAI-1 may influence the vascular response to long-term NOS inhibition by N(omega)-nitro-L-arginine methyl ester (L-NAME). METHODS AND RESULTS: We compared the temporal changes in systolic blood pressure and coronary perivascular fibrosis in PAI-1-deficient (PAI-1(-/-)) and wild-type (WT) male mice (N=6 per group). At baseline, there were no significant differences in blood pressure between groups. After initiation of L-NAME, systolic blood pressure increased in both groups at 2 weeks. Over an 8-week study period, systolic blood pressure increased to 141+/-3 mm Hg in WT animals versus 112+/-4 mm Hg in PAI-1( /-) mice (P<0.0001). The extent of coronary perivascular fibrosis increased significantly in L-NAME-treated WT mice (P<0.01 versus PAI-1(-/-) mice). Cardiac type I collagen mRNA expression was greater in control (P<0.01) and L-NAME treated PAI-1(-/-) (P<0.05) groups than in control WT mice, indicating that PAI-1 deficiency prevents the increase of collagen deposition by promoting matrix degradation. CONCLUSIONS: These findings suggest that PAI-1 deficiency alone is sufficient to protect against the structural vascular changes that accompany hypertension in the setting of long-term NOS inhibition. Direct inhibition of vascular PAI-1 activity may provide a new therapeutic strategy for the prevention of arteriosclerotic cardiovascular disease. PMID- 11502714 TI - Unusual clinical presentation of a patient with an extreme form of right ventricular dysplasia. PMID- 11502715 TI - Electrical connections between pulmonary veins in humans: evidence after radiofrequency ablation of the venoatrial junction. PMID- 11502716 TI - MRI Versus 3D echocardiography in postinterventional patients with hypertrophic obstructive cardiomyopathy. PMID- 11502717 TI - Pizza and vegetables don't stick to the endothelium. PMID- 11502718 TI - Is cardiopulmonary bypass really the cause of postoperative atrial fibrillation? PMID- 11502719 TI - Metoprolol-induced changes in myocardial (123)I-metaiodobenzylguanidine uptake in Parkinson's disease. PMID- 11502720 TI - Atherosclerosis: Is it time for a new name? PMID- 11502721 TI - AHA Conference Proceedings: Do existing databases hold the answers to clinical questions in geriatric cardiovascular disease and stroke? Executive Summary. Database Conference, January 27-30, 2000. Washington, DC, USA. PMID- 11502722 TI - Bristol scandal. PMID- 11502723 TI - In vitro characterization of the inhibition profile of loratadine, desloratadine, and 3-OH-desloratadine for five human cytochrome P-450 enzymes. AB - The purpose of this study was to evaluate loratadine, desloratadine, and 3-OH desloratadine as inhibitors of certain human liver cytochrome P-450 enzymes. Pooled human liver microsomes were used to determine whether loratadine, desloratadine, and 3-OH-desloratadine were inhibitors of cytochrome P-450 (CYP) 1A2, 2C9, 2C19, 2D6, and 3A4. Loratadine did not inhibit CYP1A2 or CYP3A4 at concentrations up to 3829 ng/ml, which is approximately 815-fold greater than the expected maximal human plasma concentration (4.7 +/- 2.7 ng/ml) following the recommended dose of 10 mg/day. Loratadine inhibited CYP2C19 and CYP2D6 with IC(50) values of approximately 0.76 microM [291 ng/ml; K(i) congruent with 0.61 microM (234 ng/ml)] and 8.1 microM [3100 ng/ml; K(i) congruent with 2.7 microM (1034 ng/ml)], respectively, which are approximately 62 and 660 times the expected loratadine therapeutic exposure concentration. Neither desloratadine nor 3-OH-desloratadine inhibited CYP1A2, CYP2C9, CYP2C19, CYP2D6, or CYP3A4 greater than 25% at concentrations of 3108 or 3278 ng/ml, respectively. These results suggest that loratadine and the active metabolites desloratadine and 3-OH desloratadine are unlikely to affect the pharmacokinetics of coadministered drugs which are metabolized by these five cytochrome P-450 enzymes. PMID- 11502724 TI - Metabolic activation of polycyclic aromatic hydrocarbons and other procarcinogens by cytochromes P450 1A1 and P450 1B1 allelic variants and other human cytochromes P450 in Salmonella typhimurium NM2009. AB - A variety of polycyclic aromatic hydrocarbons and their dihydrodiol derivatives, arylamines, heterocyclic amines, and nitroarenes, were incubated with cDNA-based recombinant (Escherichia coli or Trichoplusia ni) systems expressing different forms of human cytochrome P450 (P450 or CYP) and NADPH-P450 reductase using Salmonella typhimurium tester strain NM2009, and the resultant DNA damage caused by the reactive metabolites was detected by measuring expression of umu gene in the cells. Recombinant (bacterial) CYP1A1 was slightly more active than any of four CYP1B1 allelic variants, CYP1B1*1, CYP1B1*2, CYP1B1*3, and CYP1B1*6, in catalyzing activation of chrysene-1,2-diol, benz[a]anthracene-trans-1,2-, 3,4-, 5,6-, and 8,9-diol, fluoranthene-2,3-diol, dibenzo[a,l]pyrene, benzo[c]phenanthrene, and dibenz[a,h]anthracene and several arylamines and heterocyclic amines, whereas CYP1A1 and CYP1B1 enzymes had essentially similar catalytic specificities toward other procarcinogens, such as (+)-, (-)-, and (+/ )-benzo[a]pyrene-7,8-diol, 5-methylchrysene-1,2-diol, 7,12 dimethylbenz[a]anthracene-3,4-diol, dibenzo[a,l]pyrene-11,12-diol, benzo[b]fluoranthene-9,10-diol, benzo[c]chrysene, 5,6-dimethylchrysene-1,2-diol, benzo[c]phenanthrene-3,4-diol, 7,12-dimethylbenz[a]anthracene, benzo[a]pyrene, 5 methylchrysene, and benz[a]anthracene. We also determined activation of these procarcinogens by recombinant (T. ni) human P450 enzymes in S. typhimurium NM2009. There were good correlations between activities of procarcinogen activation by CYP1A1 preparations expressed in E. coli and T. ni cells, although basal activities with three lots of CYP1B1 in T. ni cells were very high without substrates and NADPH in our assay system. Using 14 forms of human P450s (but not CYP1B1) (in T. ni cells), we found that CYP1A2, 2C9, 3A4, and 2C19 catalyzed activation of several of polycyclic aromatic hydrocarbons at much slower rates than those catalyzed by CYP1A1 and that other enzymes, including CYP2A6, 2B6, 2C8, 2C18, 2D6, 2E1, 3A5, 3A7, and 4A11, were almost inactive in the activation of polycyclic aromatic hydrocarbons examined here. PMID- 11502725 TI - Simultaneous pharmacokinetic modeling of cocaine and its metabolites, norcocaine and benzoylecgonine, after intravenous and oral administration in rats. AB - To accurately assess the mechanism of involvement of the active metabolite norcocaine in the effects of oral cocaine, it is essential to determine the rate and extent of the formation of norcocaine. Although this study was designed specifically for this aim, it was also of interest to characterize the metabolite kinetics of benzoylecgonine for comparative purpose. We first characterized the pharmacokinetics of cocaine, norcocaine, and benzoylecgonine by the i.v. route of administration; all three drugs decayed biexponentially. These pharmacokinetic estimates were then used for determination of the formation of norcocaine and benzoylecgonine after i.v. and p.o. (20-40 mg/kg) cocaine administration. Although t(1/2alpha), and t(1/2beta) were similar across the three compounds, the values of volume of distribution in the central compartment and clearance for benzoylecgonine were much smaller than those of cocaine and norcocaine. Norcocaine was not detected following i.v. cocaine; however, serum norcocaine concentrations were as high as those of oral cocaine. Both routes of cocaine administration produced benzoylecgonine. A pharmacokinetic model for the metabolite kinetics was proposed by sequentially adding the models that most adequately described the formation of each metabolite to the model of cocaine. For oral cocaine, the absolute bioavailability was 3.48%, whereas 6.04 and 2.26% of cocaine were converted to benzoylecgonine and norcocaine, respectively, during first-pass absorption regardless of dose. Furthermore, the majority of norcocaine and 92% of benzoylecgonine were formed during the first-pass absorption, leaving 8% of benzoylecgonine produced in systemic circulation. The profile of norcocaine as a metabolite confirmed the involvement of norcocaine in cocaine's behavioral effects. PMID- 11502726 TI - Mechanism-based inactivation of CYP2C11 by diclofenac. AB - It has been known that diclofenac is biotransformed into chemically reactive metabolites, which bind covalently to liver microsomal proteins, including cytochrome P450 enzyme(s). We have investigated the ability and selectivity of diclofenac to inactivate P450 enzymes. Preincubation of microsomes of untreated rats with diclofenac in the presence of NADPH resulted in time-dependent loss of testosterone 2alpha- and 16alpha-hydroxylation activities. No effect of the preincubation was observed on ethoxyresorufin O-deethylase, pentoxyresorufin O depentylase, or testosterone 6beta-hydroxylation activity. The time-dependent decreases in testosterone 2alpha- and 16alpha-hydroxylation activities followed the pseudo-first order kinetics and were saturable with increasing diclofenac concentrations. Reduced glutathione was not capable of protecting against the decrease in the enzyme activities. These data establish that a mechanism-based inactivation of CYP2C11 occurs during the oxidative metabolism of diclofenac. The diclofenac concentrations required to achieve the half-maximal rate of inactivation (K(I)) were 3 to 4 microM, which were close to K(m) for the low-K(m) components for diclofenac 4'- and 5-hydroxylation activities (7.29 and 4.43 microM, respectively). Anti-CYP2C11 IgG inhibited diclofenac 4'- and 5 hydroxylation activities, indicating that CYP2C11 is a major isozyme responsible for these aromatic oxidations. The preincubation of microsomes with 4'- or 5 hydroxydiclofenac did not cause a decrease in testosterone 2alpha- or 16alpha hydroxylation activity, suggesting that neither of the primary metabolites is a precursor of the metabolite that inactivates CYP2C11. Therefore, a highly reactive intermediate(s) inactivating CYP2C11, probably arene-oxide, appears to be generated during the process of diclofenac 4'- and/or 5-hydroxylation. Diclofenac metabolism in human liver microsomes did not cause inactivation of CYP2C9, a major isozyme involved in diclofenac 4'-hydroxylation. Because the human microsomes have high diclofenac 4'-hydroxylation but not 5-hydroxylation activity, importance of the latter pathway in the inactivation is suggested. PMID- 11502727 TI - The use of 3-[2-(N,N-diethyl-N-methylammonium)ethyl]-7-methoxy-4-methylcoumarin (AMMC) as a specific CYP2D6 probe in human liver microsomes. AB - Recently, a novel nonfluorescent probe 3-[2-(N,N-diethyl-N-methylammonium)-ethyl] 7-methoxy-4-methylcoumarin (AMMC), which produces a fluorescent metabolite AMHC (3-[2-(N,N-diethyl-N-methylammonium)ethyl]-7-hydroxy-4-methylcoumarin) was used with microsomes containing recombinant enzymes (rCYP) to monitor CYP2D6 inhibition in a microtiter plate assay. This article describes the studies that were performed in human liver microsomes (HLM) to establish the selectivity of AMMC toward CYP2D6. Metabolism studies in HLM showed that AMMC was converted to one metabolite identified by mass spectrometry as AMHC. Kinetic studies indicated an apparent K(m) of 3 microM with a V(max) of 20 pmol/min. mg of protein for the O-demethylation reaction. The O-demethylation of AMMC in HLM was inhibited significantly in the presence of a CYP2D6 inhibitory antibody. Using a panel of various HLM preparations (n = 12), a good correlation (r(2) = 0.95) was obtained between AMMC O-demethylation and bufuralol metabolism, a known CYP2D6 substrate, but not with probes for the other major xenobiotic metabolizing CYPs. Finally, only rCYP2D6 showed detectable metabolism in experiments conducted with rCYPs using AMMC at a concentration of 1.5 microM (near K(m)). However, at a concentration of 25 microM AMMC, rCYP1A also contributed significantly to the formation of AMHC. Knowing the experimental conditions under which AMMC was selective for CYP2D6, a microtiter assay was developed to study the inhibition of various compounds in HLM using the fluorescence of AMHC as an indication of CYP2D6 activity. The inhibition potential of various chemicals was found to be comparable to those determined using the standard CYP2D6 probe, bufuralol, which requires high-performance liquid chromatography separation for the analysis of its CYP2D6-mediated 1'-hydoxylated metabolite. PMID- 11502728 TI - Metabolism of chlorpyrifos by human cytochrome P450 isoforms and human, mouse, and rat liver microsomes. AB - One of the factors determining the toxicity of chlorpyrifos (CPS), an organophosphorus (OP) insecticide, is its biotransformation. CPS can be activated by cytochrome P450 (CYP) through a desulfuration reaction to form chlorpyrifos oxon (CPO), a potent anticholinesterase. CPS can also be detoxified by CYP through a dearylation reaction. Using pooled human liver microsomes (HLM), a K(m(app)) of 30.2 microM and V(max(app)) of 0.4 nmol/min/mg of protein was obtained for desulfuration, and a K(m(app)) of 14.2 microM and a V(max(app)) of 0.7 nmol/min/mg of protein was obtained for dearylation. These activities are lower than those obtained from rat liver microsomes. Gender differences in humans were also observed with female HLM possessing greater activity than male HLM. Use of human CYP isoforms expressed in human lymphoblastoma cells demonstrated that CYP1A2, 2B6, 2C9*1, 2C19, and 3A4 are involved in CPS metabolism. CYP2B6 has the highest desulfuration activity, whereas dearylation activity is highest for 2C19. CYP3A4 has high activity for both dearylation and desulfuration. The use of phenotyped individual HLM demonstrated that predictions of metabolic activation and/or detoxication could be made based on relative amounts of CYP2B6, 2C19, and 3A4 in the microsomes. Thus, individuals with high CYP2C19 but low 3A4 and 2B6 are more active in dearylation than in desulfuration. Similarly, individuals possessing high levels of CYP2B6 and 3A4 have the greatest potential to form the activation product. These differences between individuals suggest that differential sensitivities to CPS may exist in the human population. PMID- 11502729 TI - Genetic polymorphism of cytochrome P450 3A5 in Chinese. AB - The CYP3A subfamily enzymes are the most abundant and important drug-metabolizing enzymes. Wide variation in the CYP3A5 expression was well known. Recently, G(-44) to A of CYP3AP1 was found to segregate with CYP3A5*3 defective allele. The homozygous A(-44) subjects showed low expression of CYP3A5. In Caucasian, only 9.2% of CYP3AP1 alleles were with G(-44) and associated with the wild-type CYP3A5*1 allele, which expressed CYP3A5 significantly. By using polymerase chain reaction and FauI endonuclease digestion, we found that 28% of CYP3AP1 alleles are G(-44) in 110 Chinese subjects. The frequency is 3 times higher in Chinese than in Caucasian, implying more Chinese subjects are probably extensive CYP3A5 metabolizers. In two Chinese subjects, we also found a heterozygous G(13048)gt-to G(13048)gc mutation at the intron 5 splicing donor site, leading to a splicing defect. A 6478-base pair minigene, including intron 4 to intron 7, was used for in vitro transcription. Both the wild-type and the mutated minigenes produced splicing variants. The wild-type minigene used Ggt(13050) as the splicing donor. The mutant minigene used gt(8504) in intron 4 or gt(13112) in intron 5 as the splicing donor for various splicing acceptors. The splicing defect may result in a shorter peptide or cause the frame shift. In the other two Chinese subjects, we found A(14763)-to-G mutation in exon 7, resulting in the Q200R amino acid change. The consequence of the polymorphism site has not been known. In Caucasian, there is a reported T398N polymorphism. In these Chinese subjects, we did not find polymorphism at this site. PMID- 11502730 TI - Effect of alpha-fluorination of valproic acid on valproyl-S-acyl-CoA formation in vivo in rats. AB - Studies designed to compare valproic acid (VPA) with its alpha-fluorinated derivative (F-VPA) for their abilities to form acyl-CoA thioester derivatives in vivo are described. Recent studies have shown that alpha-fluorination of a hepatotoxic metabolite of VPA (Delta(4)-VPA) resulted in a nonhepatotoxic derivative. We hypothesize that the decrease in hepatotoxicity may be related to a lack of formation of the intermediary acyl-CoA thioester. To determine the effect of alpha-fluoro substitution on acyl-CoA formation, we synthesized F-VPA and compared it with VPA for its ability to form the acyl-CoA thioester derivative in vivo in rat liver. Thus, after dosing rats with VPA or F-VPA, animals were sacrificed (0.05-, 0.5-, 1-, 2-, and 5-h postadministration) for the analysis of liver tissue. High-performance liquid chromatography (HPLC) and electrospray ionization/tandem mass spectrometry analysis of liver extracts from VPA-dosed rats showed the presence of VPA-CoA that was maximal after 0.5 h (185 nmol/g of liver) and was still measurable 5-h postadministration (90 nmol/g of liver). In agreement with our hypothesis, F-VPA did not form the corresponding acyl-CoA derivative as determined by the absence of F-VPA-CoA upon HPLC analysis of liver extracts from F-VPA-dosed rats. Further examination of liver tissue for the presence of free acids revealed that the differences in acyl-CoA formation cannot be explained by differences in VPA and F-VPA free acid concentrations. From these observations and related studies showing the lack of toxicity due to alpha-fluoro substitution, we propose that metabolism of VPA by acyl-CoA formation may mediate the hepatotoxicity of the drug. PMID- 11502731 TI - Metabolism of vanoxerine, 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3 phenylpropyl)piperazine, by human cytochrome P450 enzymes. AB - Vanoxerine (1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine; GBR12909) is a promising agent for the treatment of cocaine dependence. Knowledge of the major pathway for GBR12909 metabolism is important for prediction of the likelihood of drug-drug interactions, which may affect the therapeutic clinical outcome, when this agent is used in cocaine-dependent individuals receiving multiple drug therapy. We studied biotransformation of GBR12909 in human liver microsomes (n = 4), human hepatocytes, and microsomes containing cDNA-expressed human P450 isoforms with GBR12909 concentrations within the range of steady-state plasma concentrations detected in healthy volunteers. A high-pressure liquid chromatography assay was used to measure parent GBR12909 and its primary metabolite. GBR12909 was metabolized by human liver microsomes, hepatocytes, and cDNA-expressed human P450s to a single metabolite. Ketoconazole, a selective inhibitor of CYP3A, reduced GBR12909 biotransformation in human liver microsomes and primary hepatocytes by 92 +/- 2 and 92.4 +/- 0.4%, respectively. Quercetin (an inhibitor of CYP2C8/3A4) was a less effective inhibitor producing 62 +/- 22% inhibition in human liver microsomes and 54 +/- 35% in hepatocytes. Other P450 selective inhibitors did not decrease GBR12909 biotransformation more than 29% in either human liver microsomes or hepatocytes with the exception of chlorzoxazone (CYP2E1), which inhibited GBR12909 biotransformation by 71.4 +/- 18.5% in primary human hepatocytes. Ciprofloxacin (CYP1A2), sulfaphenazole (CYP2C9), quinidine (CYP2D6), chlorzoxazone (CYP2E1), and mephenytoin (CYP2C19) did not demonstrate statistically significant inhibition (p > 0.05) of GBR12909 biotransformation in liver microsomes. cDNA-expressed P450 3A4 metabolized GBR12909 to a greater extent than 2C8 and 2E1. These data suggest the possibility that multiple P450 isoforms may be involved in human GBR12909 metabolism but that CYP3A appears to be the major enzyme responsible for human GBR12909 biotransformation. PMID- 11502732 TI - Depentylation of the rat esophageal carcinogen, methyl-n-pentylnitrosamine, by microsomes from various human and rat tissues and by cytochrome P450 2A3. AB - Methyl-n-pentylnitrosamine (MPN) is carcinogenic for the rat esophagus. To determine organ specificity for MPN activation by human tissues, microsomes isolated from human organs (snap-frozen <6 h after death or removed surgically) were incubated with [pentyl-(3)H]MPN, and [(3)H]pentaldehyde formation was measured by high-pressure liquid chromatography of its 2,4-dinitrophenylhydrazone using radioflow assay. With 100 microM MPN, mean depentylation rates were 6.6 (liver), 2.9 to 3.8 (kidney, stomach, small intestine, and colon), and 0.4 to 1.6 (esophagus, lung, and skin) pmol of pentaldehyde/mg of protein/min. Of 14 human esophagi, four showed relatively high depentylation rates of 3.3 to 4.1 pmol/mg/min. Apparent K(m) was 80 to 160 microM (V(max), 3-15 pmol/mg/min) for three esophagi, 90 to 130 (2 livers), and 1330 (1 kidney) microM. Rat tissues showed mean depentylation rates for 100 microM MPN of 24.9 (liver), 14.5 (esophagus), 7.0 (lung), and 0.0 to 2.7 (5 other tissues) pmol/mg/min. MPN depentylation by rat cytochrome P450 2A3 showed an apparent K(m) of 8 microM (V(max), 70 pmol/nmol of P450/min) and was competitively inhibited by the CYP2A inhibitor coumarin (apparent K(i), 4 microM). Coumarin (0.4 mM) inhibited microsomal depentylation of 100 microM MPN by 37 to 62% for human esophagus, liver, kidney, and colon and for rat esophagus but not for rat liver and lung. MPN depentylation by rat esophageal microsomes increased up to 90% on adding P450 reductase. The results indicate organ-specific MPN metabolism by rat but not human esophagus. Nevertheless, the relatively high activity of four human esophagi might indicate increased susceptibility of some individuals to carcinogenesis by unsymmetrical dialkylnitrosamines. PMID- 11502733 TI - Induction of cytochromes P450 1A1 and 1B1 by emodin in human lung adenocarcinoma cell line CL5. AB - Emodin (3-methyl-1,6,8-trihydroxyanthraquinone) is an active compound of many laxative herbal drugs. The present study aimed to determine the effects of emodin on cytochrome P450 (P450)-dependent monooxygenases of human lung adenocarcinoma CL5 cells. Treatment of CL5 cells with 100 microM emodin for 24 h induced benzo[a]pyrene hydroxylation, 7-ethoxyresorufin O-deethylation, and 7 ethoxycoumarin O-deethylation activities of S9 fractions. Immunoblot analysis of CL5 S9 proteins revealed that emodin induced proteins immunorelated to P450s 1A1 and 1B1. Northern blot analysis of total cellular RNA showed that emodin induced P450s 1A1 and 1B1 mRNA levels in CL5 cells. These inductive effects on P450 monooxygenase activity, protein, and mRNA were concentration- and time-dependent. Addition of emodin to CL5 cell microM S9 inhibited its 7-ethoxycoumarin O deethylation activity. Treatment of CL5 cells with 10 microM 3-methylcholanthrene for 24 h induced monooxygenase activity and P450s 1A1 and 1B1 proteins and mRNA levels. Treatment of the lung cells with 100 microM emodin or purpurin (1,2,4 trihydroxyanthraquinone) for 24 h produced greater induction of P450s 1A1 and 1B1 mRNA than did anthraflavic acid (2,6-dihydroxyanthraquinone) or anthraquinone. The emodin treatment induced P450s 1A1 and 1B1 mRNA in human lung carcinoma NCI H322 and breast cancer MCF-7 cells. Emodin induced P450 1A1, but not 1B1, mRNA in human hepatoma HepG2 cells. The present study demonstrates that emodin is an inducer of P450s 1A1 and 1B1 protein and mRNA in human lung adenocarcinoma CL5 cells. Modulation of P450 by emodin may be an important factor affecting metabolism and toxicity of the hydroxyanthraquinone in humans. PMID- 11502734 TI - Role of induction of specific hepatic cytochrome P450 isoforms in epoxidation of 4-vinylcyclohexene. AB - 4-Vinyl-1-cyclohexene (VCH) is ovotoxic in B6C3F(1) mice but not in Fischer-344 rats, which can be partially attributed to greater formation of toxic epoxides from VCH in mice compared with rats. Since repeated exposure to VCH is necessary to cause ovotoxicity in mice, it is important to determine whether repeated exposure results in induction of cytochrome P450 (CYP) enzymes involved in its bioactivation. Hepatic microsomes prepared from mice or rats treated repeatedly with VCH demonstrated significantly increased VCH bioactivation in vitro, as assessed by VCH-1,2-epoxide, VCH-7,8-epoxide, or vinylcyclohexene diepoxide (VCD) formation. Mice and rats were then dosed with VCH, VCH-1,2-epoxide, or VCD for 10 days and measured for increases in hepatic microsomal CYP levels or activities. Total hepatic CYP levels were elevated only in microsomes from mice pretreated with VCH or VCH-1,2-epoxide. Immunoblotting analysis of microsomes from VCH treated rodents revealed elevated levels of CYP2A and CYP2B in mice but not rats. VCH-1,2-epoxide pretreatment also increased CYP2B levels in the mouse. Activities toward specific substrates for CYP2A and CYP2B (coumarin and pentoxyresorufin, respectively) confirmed that VCH and VCH-1,2-epoxide pretreatments resulted in increased catalytic activities of CYP2A and CYP2B in the mouse but not the rat. Pretreatment with phenobarbital, a known inducer of CYP2A and CYP2B, increased VCH bioactivation in both species. Interestingly, metabolism studies with human CYP "Supersomes" reveal that, of eight isoforms tested, only human CYP2E1 and CYP2B6 were capable of significantly catalyzing VCH epoxidation, whereas CYP2B6, CYP2A6, CYP2E1, and CYP3A4 were capable of catalyzing the epoxidation of the monoepoxides. PMID- 11502735 TI - Cytochrome P450 induction in rat hepatocytes assessed by quantitative real-time reverse-transcription polymerase chain reaction and the RNA invasive cleavage assay. AB - The acceleration of drug discovery due to combinatorial chemistry and high throughput screening methods has increased the numbers of candidate pharmaceuticals entering the drug development phase, and the capability to accurately predict whether drug candidates will induce various members of the drug-metabolizing cytochrome P450 (CYP) enzyme superfamily is currently of great interest to the pharmaceutical industry. In the present study, we describe the rapid and reliable analysis of CYP induction in a readily obtained model system (cultured rat hepatocytes) using both real-time quantitative reverse transcription-polymerase chain reaction (real-time RT-PCR) and the RNA invasive cleavage assay. The levels of members in the three primary inducible rat CYP subfamilies (CYP1A1, CYP2B1/2, and CYP3A1) were analyzed in untreated and induced (beta-naphthoflavone, phenobarbital, and hydrocortisone) hepatocyte cultures under various media conditions to screen for optimal CYP induction profiles. The fold inductions measured by real-time RT-PCR and the RNA invasive cleavage assay were also compared with enzyme activity measurements in parallel cultures using liquid chromatography/double mass spectrometry-based assays, and the sensitivity and the specificity of the two RNA analysis methods were compared. Using these techniques, various culture conditions were examined for optimizing induction of the three CYP subfamily members. Both real-time RT-PCR and the RNA invasive cleavage assay prove to be effective methods for determining the effects of drugs on specific CYPs in primary rat hepatocytes. PMID- 11502736 TI - Calcium binding properties of gamma-crystallin: calcium ion binds at the Greek key beta gamma-crystallin fold. AB - The beta- and gamma-crystallins are closely related lens proteins that are members of the betagamma-crystallin superfamily, which also include many non-lens members. Although beta-crystallin is known to be a calcium-binding protein, this property has not been reported in gamma-crystallin. We have studied the calcium binding properties of gamma-crystallin, and we show that it binds 4 mol eq of calcium with a dissociation constant of 90 microm. It also binds the calcium mimic spectral probes, terbium and Stains-all. Calcium binding does not significantly influence protein secondary and tertiary structures. We present evidence that the Greek key crystallin fold is the site for calcium ion binding in gamma-crystallin. Peptides corresponding to Greek key motif of gamma crystallin (42 residues) and their mutants were synthesized and studied for calcium binding. These peptides adopt beta-sheet conformation and form aggregates producing beta-sandwich. Our results with peptides show that, in Greek key motif, the amino acid adjacent to the conserved aromatic corner in the "a" strand and three amino acids of the "d" strand participate in calcium binding. We suggest that the betagamma superfamily represents a novel class of calcium-binding proteins with the Greek key betagamma-crystallin fold as potential calcium binding sites. These results are of significance in understanding the mechanism of calcium homeostasis in the lens. PMID- 11502737 TI - Control of myoblast proliferation with a synthetic ligand. AB - Skeletal myoblast grafts can form contractile tissue to replace scar and repair injured myocardium. Although potentially therapeutic, generating reproducible and sufficiently large grafts remains a challenge. To control myoblast proliferation in situ, we created a chimeric receptor composed of a modified FK506-binding protein (F36V) fused with the fibroblast growth factor receptor-1 cytoplasmic domain. Mouse MM14 myoblasts were transfected with this construct and treated with AP20187, a dimeric F36V ligand, to induce receptor dimerization. Transfected myoblasts proliferated in response to dimerizer (comparable with basic fibroblast growth factor (bFGF) treatment), whereas the dimerizer had no effect on non transfected cells. Similar to bFGF treatment, dimerizer treatment blocked myotube formation and myosin heavy chain expression and stimulated mitogen-activated protein (MAP) kinase phosphorylation in transfected cells. Non-transfected cells differentiated normally and showed no MAP kinase phosphorylation with dimerizer treatment. Furthermore, myoblasts treated with dimerizer for 30 days in culture reduced MAP kinase phosphorylation, withdrew from the cell cycle, and differentiated normally upon drug withdrawal, demonstrating reversibility of the effect. Thus, forced dimerization of the fibroblast growth factor receptor-1 cytoplasmic domain reproduces critical aspects of bFGF signaling in myoblasts. We hypothesize that in vivo administration of AP20187 following myoblast grafting may allow control over graft size and ultimately improve cardiac function. PMID- 11502738 TI - Angiotensin II-induced transcriptional activation of the cyclin D1 gene is mediated by Egr-1 in CHO-AT(1A) cells. AB - Cyclin D1 protein expression is regulated by mitogenic stimuli and is a critical component in the regulation of G(1) to S phase progression of the cell cycle. Angiotensin II (Ang II) binds to specific G protein-coupled receptors and is mitogenic in Chinese hamster ovary cells stably expressing the rat vascular Ang II type 1A receptor (CHO-AT(1A)). We recently reported that in these cells, Ang II induced cyclin D1 promoter activation and protein expression in a phosphatidylinositol 3-kinase (PI3K)-, SHP-2-, and mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK)-dependent manner (Guillemot, L., Levy, A., Zhao, Z. J., Bereziat, G., and Rothhut, B. (2000) J. Biol. Chem. 275, 26349-26358). In this report, transfection studies using a series of deleted cyclin D1 promoters revealed that two regions between base pairs (bp) -136 and -96 and between bp -29 and +139 of the human cyclin D1 promoter contained regulatory elements required for Ang II-mediated induction. Mutational analysis in the -136 to -96 bp region provided evidence that a Sp1/early growth response protein (Egr) motif was responsible for cyclin D1 promoter activation by Ang II. Gel shift and supershift studies showed that Ang II-induced Egr-1 binding involved de novo protein synthesis and correlated well with Egr-1 promoter activation. Both U0126 (an inhibitor of the MAPK/ERK kinase MEK) and wortmannin (an inhibitor of PI3K) abrogated Egr-1 endogenous expression and Egr-1 promoter activity induced by Ang II. Moreover, using a co-transfection approach, we found that Ang II induction of Egr-1 promoter activity was blocked by dominant-negative p21(ras), Raf-1, and tyrosine phosphatase SHP-2 mutants. Identical effects were obtained when inhibitors and dominant negative mutants were tested on the -29 to +139 bp region of the cyclin D1 promoter. Taken together, these findings demonstrate that Ang II-induced cyclin D1 up-regulation is mediated by the activation and specific interaction of Egr-1 with the -136 to 96 bp region of the cyclin D1 promoter and by activation of the -29 to +139 bp region, both in a p21(ras)/Raf-1/MEK/ERK-dependent manner, and also involves PI3K and SHP-2. PMID- 11502739 TI - Four PSM/SH2-B alternative splice variants and their differential roles in mitogenesis. AB - An SH2 domain originally termed SH2-B had been identified as a direct cellular binding target of a number of mostly mitogenic receptors. The complete cellular protein, termed PSM, and respective sequence variants share additional Pro-rich and PH regions, as well as similarities with APS and Lnk. A role of these mediators has been implicated in signaling pathways found downstream of growth hormone receptor and receptor tyrosine kinases, including the insulin, insulin like growth factor-I (IGF-I), platelet-derived growth factor (PDGF), nerve growth factor, hepatocyte growth factor, and fibroblast growth factor receptors. As a result of this report a total of four PSM/SH2-B sequence variants termed alpha, beta, gamma, and delta have now been identified in the mouse and have been compared with the available rat and human sequences. Variant differences are based on alternative splicing and define distinct last exons 7, 8, and 9 that result in reading frameshifts and unique carboxyl-terminal amino acid sequences. Variant sequences have been identified from cDNA libraries and directly by reverse transcription-polymerase chain reaction. Sequence analysis predicts four distinctly sized protein products that have been demonstrated after cDNA expression. All were found phosphorylated on tyrosine specifically in response to IGF-I and PDGF stimulation. cDNA expression of the four variants caused variant dependent levels of stimulation of IGF-I- and PDGF-induced mitogenesis. The most pronounced increase in mitogenesis was consistently observed for the gamma variant followed by delta, alpha, and beta with decreasing responses. In contrast, the mitogenic response to epidermal growth factor consistently remained unaffected. The variants are expressed in most mouse tissues, typically, most strongly in pairs of alpha and delta or beta and gamma. Our findings implicate differential roles of the PSM/SH2-B splice variants in specific mitogenic signaling pathways. PMID- 11502740 TI - Neuregulin-heparan-sulfate proteoglycan interactions produce sustained erbB receptor activation required for the induction of acetylcholine receptors in muscle. AB - Neuregulins bind to and activate members of the EGF receptor family of tyrosine kinases that initiate a signaling cascade that induces acetylcholine receptor synthesis in the postsynaptic membrane of neuromuscular synapses. In addition to an EGF-like domain, sufficient for receptor binding and tyrosine auto phosphorylation, many spliced forms also have an IG-like domain that binds HSPGs and maintains a high concentration of neuregulin at synapses. Here, we show that the IG-like domain functions to keep the EGF-like domain at sufficiently high concentrations for a sufficiently long period of time necessary to induce acetylcholine receptor gene expression in primary chick myotubes. Using recombinant neuregulins with and without the IG-like domain, we found that IG like domain binding to endogenous HSPGs produces a 4-fold increase in receptor phosphorylation. This enhancement of activity was blocked by soluble heparin or by pretreatment of muscle cells with heparitinase. We show that at least 12-24 h of neuregulin exposure was required to turn on substantial acetylcholine receptor gene expression and that the erbB receptors need to be kept phosphorylated during this time. The need for sustained erbB receptor activation may be the reason why neuregulins are so highly concentrated in the extracellular matrix of synapses. PMID- 11502741 TI - An isoform of the coactivator AIB1 that increases hormone and growth factor sensitivity is overexpressed in breast cancer. AB - The AIB1 (amplified in breast cancer 1) protein is a coactivator that potentiates the transcriptional activity of nuclear hormone receptors, and its gene is amplified in a subset of human breast cancers. Here we report a splice variant of AIB1 mRNA that lacks the exon 3 sequence. We determined that the AIB-Delta3 mRNA encoded a 130-kDa protein that lacks the NH(2)-terminal basic helix-loop-helix and a portion of the PAS (Per-Arnt-Sim homology) dimerization domain. The 130-kDa protein was detected in MCF-7 breast cancer cells at levels that were 5-10% of the full-length protein, whereas in non-transformed mammary epithelium lines, the AIB-Delta3 protein was present at significantly lower levels compared with the full-length AIB1. Consistent with this finding, the abundance of AIB1-Delta3 mRNA was increased in human breast cancer specimens relative to that in normal breast tissue. To determine whether there were phenotypic changes associated with the overexpression of the AIB-Delta3 isoform, we performed functional reporter gene assays. These revealed that the ability of AIB1-Delta3 to promote transcription mediated by the estrogen or progesterone receptors was significantly greater than that of the full-length protein. Surprisingly, the AIB1-Delta3 isoform was also more effective than AIB1 in promoting transcription induced by epidermal growth factor. Overexpression of AIB1-Delta3 may thus play an important role in sensitizing breast tumor cells to hormone or growth factor stimulation. PMID- 11502742 TI - Protein kinase C regulates the phosphorylation and cellular localization of occludin. AB - Occludin is an integral membrane phosphoprotein specifically associated with tight junctions, contributing to the structure and function of this intercellular seal. Occludin function is thought to be regulated by phosphorylation, but no information is available on the molecular pathways involved. In the present study, the involvement of the protein kinase C pathway in the regulation of the phosphorylation and cellular distribution of occludin has been investigated. Phorbol 12-myristate 13-acetate and 1,2-dioctanoylglycerol induced the rapid phosphorylation of occludin in Madin-Darby canine kidney cells cultured in low extracellular calcium medium with a concomitant translocation of occludin to the regions of cell-cell contact. The extent of occludin phosphorylation as well as its incorporation into tight junctions induced by protein kinase C activators or calcium switch were markedly decreased by the protein kinase C inhibitor GF 109203X. In addition, in vitro experiments showed that the recombinant COOH terminal domain of murine occludin could be phosphorylated by purified protein kinase C. Ser(338) of occludin was identified as an in vitro protein kinase C phosphorylation site using peptide mass fingerprint analysis and electrospray ionization tandem mass spectroscopy. These findings indicate that protein kinase C is involved in the regulation of occludin function at tight junctions. PMID- 11502743 TI - Multiple phosphorylation sites of DNA polymerase alpha-primase cooperate to regulate the initiation of DNA replication in vitro. AB - DNA polymerase alpha-primase (pol-prim) is the only enzyme that can start DNA replication de novo. The 180-kDa (p180) and 68-kDa (p68) subunits of the human four-subunit enzyme are phosphorylated by Cyclin-dependent kinases (Cdks) in a cell cycle-dependent manner. Cyclin A-Cdk2 physically interacts with pol-prim and phosphorylates N-terminal amino acids of the p180 and the p68 subunits, leading to an inhibition of pol-prim in initiating cell-free SV40 DNA replication. Mutation of conserved putative Cdk phosphorylation sites in the N terminus of human p180 and p68 reduced their phosphorylation by Cyclin A-Cdk2 in vitro. In contrast to wild-type pol-prim these mutants were no longer inhibited by Cyclin A Cdk2 in the initiation of viral DNA replication. Importantly, rather than inhibiting it, Cyclin A-Cdk2 stimulated the initiation activity of pol-prim containing a triple N-terminal alanine mutant of the p180 subunit. Together these results suggest that Cyclin A-Cdk2 executes both stimulatory and inhibitory effects on the activity of pol-prim in initiating DNA replication. PMID- 11502744 TI - The inhibitory gamma subunit of the type 6 retinal cyclic guanosine monophosphate phosphodiesterase is a novel intermediate regulating p42/p44 mitogen-activated protein kinase signaling in human embryonic kidney 293 cells. AB - The inhibitory gamma subunits of the retinal rod and cone photoreceptor type 6 retinal cyclic guanosine monophosphate phosphodiesterase (PDEgamma) are expressed in non-retinal tissues. Here, we show that PDEgamma interacts with the G-protein coupled receptor kinase 2 signaling system to regulate the epidermal growth factor- and thrombin-dependent stimulation of p42/p44 mitogen-activated protein kinase in human embryonic kidney 293 cells. This is based upon several lines of evidence. First, the transfection of cells with an antisense rod PDEgamma plasmid construct, which reduced endogenous rod PDEgamma expression, ablated the epidermal growth factor- and thrombin-dependent stimulation of p42/p44 mitogen activated protein kinase. Second, the transfection of cells with recombinant rod or cone PDEgamma and/or G-protein-coupled receptor kinase 2 increased the stimulation of p42/p44 mitogen-activated protein kinase by epidermal growth factor or thrombin. In contrast, a G-protein-coupled receptor kinase 2 phosphorylation-resistant rod PDEgamma mutant failed to increase the epidermal growth factor- or thrombin-dependent stimulation of p42/p44 mitogen-activated protein kinase and, in fact, functioned as a dominant negative. Thrombin also stimulated the association of endogenous rod PDEgamma with dynamin II, which was increased in cells transfected with rod PDEgamma or G-protein-coupled receptor kinase 2. Dynamin II plays a critical role in regulating endocytosis of receptor signal complexes required for activation of p42/p44 mitogen-activated protein kinase. Therefore, PDEgamma may have an important role in promoting endocytosis of receptor signal complexes leading to the activation of p42/p44 mitogen activated protein kinase. We conclude that PDEgamma is an entirely novel intermediate regulating mitogenic signaling from both receptor tyrosine kinase and G-protein-coupled receptors in human embryonic kidney 293 cells. PMID- 11502745 TI - Signaling through extracellular signal-regulated kinase is required for spermatogonial proliferative response to stem cell factor. AB - In vitro addition of stem cell factor (SCF) to c-kit-expressing A(1)-A(4) spermatogonia from prepuberal mice stimulates their progression into the mitotic cell cycle and significantly reduces apoptosis in these cells. SCF addition results in a transient activation of extracellular signal-regulated kinases (Erk)1/2 as well as of phosphatidylinositol 3-kinase (PI3K)-dependent Akt kinase. These events are followed by a rapid re-distribution of cyclin D3, which becomes predominantly nuclear, whereas its total cellular amount does not change. Nuclear accumulation of cyclin D3 is coupled to transient activation of the associated kinase activity, assayed using the retinoblastoma protein (Rb) as a substrate. These events were followed by a transient accumulation of cyclin E, stimulation of the associated histone H1-kinase activity, a delayed accumulation of cyclin A2, and Rb hyper-phosphorylation. All the events associated with SCF-induced cell cycle progression are inhibited by the addition of either a PI3K inhibitor or a mitogen-activated protein-kinase kinase (MEK) inhibitor, indicating that both MEK and PI3K are essential for c-kit-mediated proliferative response. On the contrary, the anti-apoptotic effect of SCF is not influenced by the separate addition of either MEK or PI3K inhibitors. Thus, SCF effects on mitogenesis and survival in c-kit expressing spermatogonia rely on different signal transduction pathways. PMID- 11502746 TI - Phosphorylation of Ser(19) alters the conformation of tyrosine hydroxylase to increase the rate of phosphorylation of Ser(40). AB - The effect of phosphorylation on the shape of tyrosine hydroxylase (TH) was studied directly using gel filtration and indirectly using electrospray ionization mass spectrometry. Phosphorylation of Ser(19) and Ser(40) produced a TH molecule with a more open conformation than the non-phosphorylated form. The conformational effect of Ser(19) phosphorylation is less pronounced than that of the Ser(40) phosphorylation. The effect of Ser(19) and Ser(40) phosphorylation appears to be additive. Binding of dopamine produced a more compact form when compared with the non-dopamine-bound TH. The interdependence of Ser(19) and Ser(40) phosphorylation was probed using electrospray ionization mass spectrometry. The rate constants for the phosphorylation of Ser(19) and Ser(40) were determined by electrospray ionization mass spectrometry using a consecutive reaction model. The rate constant for the phosphorylation of Ser(40) is approximately 2- to 3-fold higher if Ser(19) is already phosphorylated. These results suggest that phosphorylation of Ser(19) alters the conformation of tyrosine hydroxylase to allow increased accessibility of Ser(40) to kinases. PMID- 11502747 TI - Conformations of the active and inactive states of opsin. AB - The signaling state metarhodopsin II of the visual pigment rhodopsin decays to the apoprotein opsin and all-trans retinal, which are then regenerated to rhodopsin by the visual cycle. Opsin is known to have at neutral pH only a small residual constitutive activity toward its G protein transducin, which is thought to play a considerable role in light adaptation (bleaching desensitization). In this study we show with Fourier-transform infrared spectroscopy that after metarhodopsin II decay, opsin exists in two conformational states that are in a pH-dependent equilibrium at 30 degrees C with a pK of 4.1 in the presence of hydroxylamine scavenging the endogenous all-trans retinal. Despite the lack of the native agonist in its binding pocket, the low pH opsin conformation is very similar to that of metarhodopsin II and is likewise stabilized by peptides derived from rhodopsin's cognate G protein, transducin. The high pH form, on the other hand, has some conformational similarity to the inactive metarhodopsin I state. We therefore conclude that the opsin apoprotein displays intrinsic conformational states that are merely modulated by bound all-trans retinal. PMID- 11502748 TI - The Dbl homology domain of BCR is not a simple spacer in P210BCR-ABL of the Philadelphia chromosome. AB - The Dbl homology (DH) domain of BCR in P210BCR-ABL (P210/WT) has been thought to have a negative effect on the activation of BCR-ABL because P185BCR-ABL, in which this region is physically deleted, has stronger biochemical and biological activities. To study the role of the DH domain of BCR in the background of P210/WT, the region was replaced with homologous sequences derived from Dbl (P210/Dbl) or CDC24 (P210/CDC24) or with irrelevant sequences from LacZ (P210/LacZ) or luciferase (P210/Luci). Surprisingly, the abilities to transform Rat1 cells or mouse bone marrow cells and induce growth factor independence in interleukin 3-dependent mouse Ba/F3 cells were retained only in P210/Dbl. However, even P210/Dbl could not achieve the wild type level of surviving potential against genotoxins in Rat1 cells and in Ba/F3 cells. Activation of Akt correlated with the biological changes in Rat1 cells but did not correlate with the biological changes in Ba/F3 cells. The DH domain was not tyrosine phosphorylated in vitro, nor could we find any differences in peptide mapping between in vitro phosphorylated P210/WT and P210/Dbl. Although functions of the DH domain remain to be discovered, we propose that the DH domain makes positive contributions to P210BCR-ABL. PMID- 11502749 TI - The basic helix-loop-helix domain of the aryl hydrocarbon receptor nuclear transporter (ARNT) can oligomerize and bind E-box DNA specifically. AB - The aryl hydrocarbon receptor nuclear transporter (ARNT) is a basic helix-loop helix (bHLH) protein that contains a Per-Arnt-Sim (PAS) domain. ARNT heterodimerizes in vivo with other bHLH PAS proteins to regulate a number of cellular activities, but a physiological role for ARNT homodimers has not yet been established. Moreover, no rigorous studies have been done to characterize the biochemical properties of the bHLH domain of ARNT that would address this issue. To begin this characterization, we chemically synthesized a 56-residue peptide encompassing the bHLH domain of ARNT (residues 90-145). In the absence of DNA, the ARNT-bHLH peptide can form homodimers in lower ionic strength, as evidenced by dynamic light scattering analysis, and can bind E-box DNA (CACGTG) with high specificity and affinity, as determined by fluorescence anisotropy. Dimers and tetramers of ARNT-bHLH are observed bound to DNA in equilibrium sedimentation and dynamic light scattering experiments. The homodimeric peptide also undergoes a coil-to-helix transition upon E-box DNA binding. Peptide oligomerization and DNA affinity are strongly influenced by ionic strength. These biochemical and biophysical studies on the ARNT-bHLH reveal its inherent ability to form homodimers at concentrations supporting a physiological function and underscore the significant biochemical differences among the bHLH superfamily. PMID- 11502750 TI - FAS activation induces dephosphorylation of SR proteins; dependence on the de novo generation of ceramide and activation of protein phosphatase 1. AB - The search for potential targets for ceramide action led to the identification of ceramide-activated protein phosphatases (CAPP). To date, two serine/threonine protein phosphatases, protein phosphatase 2A (PP2A) and protein phosphatase 1 (PP1), have been demonstrated to function as ceramide-activated protein phosphatases. In this study, we show that treatment with either anti-FAS IgM (CH 11) (150 ng/ml) or exogenous d-(e)-C(6-)ceramide (20 microm) induces the dephosphorylation of the PP1 substrates, serine/arginine-rich (SR) proteins, in Jurkat acute leukemia T-cells. The serine/threonine protein phosphatase inhibitor, calyculin A, but not the PP2A-specific inhibitor, okadaic acid, inhibited both FAS- and ceramide-induced dephosphorylation of SR proteins. Anti FAS IgM treatment of Jurkat cells led to a significant increase in levels of endogenous ceramide beginning at 2 h with a maximal increase of 10-fold after 7 h. A 2-h pretreatment of Jurkat cells with fumonisin B(1) (100 microm), a specific inhibitor of CoA-dependent ceramide synthase, blocked 80% of the ceramide generated and completely inhibited the dephosphorylation of SR proteins in response to anti-FAS IgM. Moreover, pretreatment of Jurkat cells with myriocin, a specific inhibitor of serine-palmitoyl transferase (the first step in de novo synthesis of ceramide), also blocked FAS-induced SR protein dephosphorylation, thus demonstrating a role for de novo ceramide. These results were further supported using A549 lung adenocarcinoma cells treated with d-(e) C(6-)ceramide. Dephosphorylation of SR proteins was inhibited by fumonisin B(1) and by overexpression of glucosylceramide synthase; again implicating endogenous ceramide generated de novo in regulating the dephosphorylation of SR proteins in response to FAS activation. These results establish a specific intracellular pathway involving both de novo ceramide generation and activation of PP1 to mediate the effects of FAS activation on SR proteins. PMID- 11502751 TI - Identification and characterization of a novel inositol hexakisphosphate kinase. AB - The inositol pyrophosphate disphosphoinositol pentakisphosphate (PP InsP(3)/InsP(7)) is formed in mammals by two recently cloned inositol hexakiphosphate kinases, InsP(6)K1 and InsP(6)K2 (Saiardi, A., Erdjument-Bromage, H., Snowman, A. M., Tempst, P., and Snyder, S. H. (1999) Curr. Biol. 9, 1323 1326). We now report the identification, cloning, and characterization of a third InsP(7) forming enzyme designated InsP(6)K3. InsP(6)K3 displays 50 and 45% sequence identity to InsP(6)K1 and InsP(6)K2, respectively, with a smaller mass (46 kDa) and a more basic character than the other two enzymes. InsP(6)K3 is most enriched in the brain where its localization resembles InsP(6)K1 and InsP(6)K2. Intracellular disposition discriminates the three enzymes with InsP(6)K2 being exclusively nuclear, InsP(6)K3 predominating in the cytoplasm, and InsP(6)K1 displaying comparable nuclear and cytosolic densities. PMID- 11502752 TI - Transforming growth factor-beta repression of matrix metalloproteinase-1 in dermal fibroblasts involves Smad3. AB - Enhanced production of matrix metalloproteinase-1 (MMP-1, collagenase-1) is implicated in pathological tissue destruction. Transforming growth factor-beta (TGF-beta) prevents cytokine-induced MMP-1 gene expression in fibroblasts. In these studies, we examined the hypothesis that repression of MMP-1 may be mediated through the Smad signaling pathway. The results showed that Smad3 and Smad4, but not Smad1 or Smad2, mimicked the inhibitory effect of TGF-beta and abrogated interleukin-1beta (IL-1beta)-induced stimulation of MMP-1 promoter activity and NFkappaB-specific gene transcription in dermal fibroblasts. Experiments with truncation mutants indicated that both MH1 and MH2 domains of Smad3 were necessary for inhibitory activity. Dominant negative mutants of Smad3 or Smad4 and antagonistic Smad7, which disrupts ligand-induced Smad3 phosphorylation, abrogated the repression of MMP-1 transcription by TGF-beta. Similar results were obtained using immunoblot and Northern analysis. Furthermore, TGF-beta failed to repress MMP-1 promoter activity in Smad3 deficient murine embryonic fibroblasts. These results implicated cellular Smads in mediating the inhibitory effects of TGF-beta. Overexpression of the transcriptional co-activator p300, but not its histone acetyltransferase (HAT) deficient mutant, was able to relieve repression of MMP-1 gene expression, suggesting that Smad-dependent inhibition may be due to increased competition between Smad proteins and IL-1beta signaling pathways for limiting amounts of cellular p300. Together, these results demonstrate that MMP-1 is a target for negative regulation by TGF-beta through cellular Smad3 and Smad4. Smad-mediated repression of MMP-1 gene expression may be important for preventing excessive matrix degradation induced by inflammatory cytokines; disruption of Smad signaling, as occurs in certain cancer cells, may thus be causally linked to uncontrolled tissue destruction mediated through MMP-1. PMID- 11502758 TI - LAD-1, the Caenorhabditis elegans L1CAM homologue, participates in embryonic and gonadal morphogenesis and is a substrate for fibroblast growth factor receptor pathway-dependent phosphotyrosine-based signaling. AB - This study shows that L1-like adhesion (LAD-1), the sole Caenorhabditis elegans homologue of the L1 family of neuronal adhesion molecules, is required for proper development of the germline and the early embryo and embryonic and gonadal morphogenesis. In addition, the ubiquitously expressed LAD-1, which binds to ankyrin-G, colocalizes with the C. elegans ankyrin, UNC-44, in multiple tissues at sites of cell-cell contact. Finally, we show that LAD-1 is phosphorylated in a fibroblast growth factor receptor (FGFR) pathway-dependent manner on a tyrosine residue in the highly conserved ankyrin-binding motif, FIGQY, which was shown previously to abolish the L1 family of cell adhesion molecule (L1CAM) binding to ankyrin in cultured cells. Immunofluorescence studies revealed that FIGQY tyrosine-phosphorylated LAD-1 does not colocalize with nonphosphorylated LAD-1 or UNC-44 ankyrin but instead is localized to sites that undergo mechanical stress in polarized epithelia and axon-body wall muscle junctions. These findings suggest a novel ankyrin-independent role for LAD-1 related to FGFR signaling. Taken together, these results indicate that L1CAMs constitute a family of ubiquitous adhesion molecules, which participate in tissue morphogenesis and maintaining tissue integrity in metazoans. PMID- 11502759 TI - A conserved alpha-herpesvirus protein necessary for axonal localization of viral membrane proteins. AB - Pseudorabies virus, an alpha-herpesvirus, is capable of infecting the nervous system and spreading between synaptically connected neurons in diverse hosts. At least three viral membrane proteins (gE, gI, and Us9) are necessary for the spread of infection from presynaptic to postsynaptic neurons (anterograde spread) in infected rodents. To understand how these proteins effect anterograde spread between neurons, we analyzed the subcellular localization of viral proteins after infection of cultured rat sympathetic neurons with wild-type or mutant viruses. After Us9-null mutant infections but not gE-null mutant infections, only a subset of the viral structural proteins had entered axons. Surprisingly, capsid and tegument proteins but not viral membrane proteins were detected in axons. The spread of Us9 missense mutants in the rodent nervous system correlated with the amount of viral membrane proteins localized to axons. We conclude that the Us9 membrane protein controls axonal localization of diverse viral membrane proteins but not that of capsid or tegument proteins. The data support a model where virion subassemblies but not complete virions are transported in the axon. Our results provide new insight into the process of virion assembly and exit from neurons that leads to directional spread of herpesviruses in the nervous system. PMID- 11502760 TI - Lipid raft microdomain compartmentalization of TC10 is required for insulin signaling and GLUT4 translocation. AB - Recent studies indicate that insulin stimulation of glucose transporter (GLUT)4 translocation requires at least two distinct insulin receptor-mediated signals: one leading to the activation of phosphatidylinositol 3 (PI-3) kinase and the other to the activation of the small GTP binding protein TC10. We now demonstrate that TC10 is processed through the secretory membrane trafficking system and localizes to caveolin-enriched lipid raft microdomains. Although insulin activated the wild-type TC10 protein and a TC10/H-Ras chimera that were targeted to lipid raft microdomains, it was unable to activate a TC10/K-Ras chimera that was directed to the nonlipid raft domains. Similarly, only the lipid raft localized TC10/ H-Ras chimera inhibited GLUT4 translocation, whereas the TC10/K Ras chimera showed no significant inhibitory activity. Furthermore, disruption of lipid raft microdomains by expression of a dominant-interfering caveolin 3 mutant (Cav3/DGV) inhibited the insulin stimulation of GLUT4 translocation and TC10 lipid raft localization and activation without affecting PI-3 kinase signaling. These data demonstrate that the insulin stimulation of GLUT4 translocation in adipocytes requires the spatial separation and distinct compartmentalization of the PI-3 kinase and TC10 signaling pathways. PMID- 11502761 TI - The AP2 binding site of synaptotagmin 1 is not an internalization signal but a regulator of endocytosis. AB - One characteristic linking members of the synaptotagmin family to endocytosis is their ability to bind the heterotetrameric AP2 complex via their C2B domain. By using CD4/synaptotagmin 1 chimeras, we found that the internalization signal of synaptotagmin 1 lies at the extreme COOH-terminus of the protein and can function in the absence of the C2B domain that contains the AP2 binding site. However, although not essential for internalization, the C2B domain of synaptotagmin 1 appeared to control the recognition of the internalization motif. By mutagenesis, two sites have been identified that modify regulation by the C2B domain in the neuroendocrine PC12 cell line. Mutation of a dilysine motif in the beta sandwich core of the domain eliminates endocytosis. This site is known to be a site of protein-protein interaction. Mutations in the calcium binding region, or in its close proximity, also affect internalization in PC12 cells. In fibroblasts, the C2B domain inhibits the COOH-terminal internalization signal, resulting in an absence of internalization in those cells. Thus, internalization of synaptotagmin 1 is controlled by the presence of a latent internalization signal in the COOH terminal region and a regulatory region in the C2B domain. We propose that internalization of synaptotagmin 1 is regulated in this way to allow it to couple the processes of endocytosis and calcium-mediated exocytosis in cells of the neuroendocrine lineage. PMID- 11502762 TI - Contraction and polymerization cooperate to assemble and close actomyosin rings around Xenopus oocyte wounds. AB - Xenopus oocytes assemble an array of F-actin and myosin 2 around plasma membrane wounds. We analyzed this process in living oocytes using confocal time-lapse (four-dimensional) microscopy. Closure of wounds requires assembly and contraction of a classic "contractile ring" composed of F-actin and myosin 2. However, this ring works in concert with a 5-10-microm wide "zone" of localized actin and myosin 2 assembly. The zone forms before the ring and can be uncoupled from the ring by inhibition of cortical flow and contractility. However, contractility and the contractile ring are required for the stability and forward movement of the zone, as revealed by changes in zone dynamics after disruption of contractility and flow, or experimentally induced breakage of the contractile ring. We conclude that wound-induced contractile arrays are provided with their characteristic flexibility, speed, and strength by the combined input of two distinct components: a highly dynamic zone in which myosin 2 and actin preferentially assemble, and a stable contractile actomyosin ring. PMID- 11502763 TI - The discrepancy between presenilin subcellular localization and gamma-secretase processing of amyloid precursor protein. AB - We investigated the relationship between PS1 and gamma-secretase processing of amyloid precursor protein (APP) in primary cultures of neurons. Increasing the amount of APP at the cell surface or towards endosomes did not significantly affect PS1-dependent gamma-secretase cleavage, although little PS1 is present in those subcellular compartments. In contrast, almost no gamma-secretase processing was observed when holo-APP or APP-C99, a direct substrate for gamma-secretase, were specifically retained in the endoplasmic reticulum (ER) by a double lysine retention motif. Nevertheless, APP-C99-dilysine (KK) colocalized with PS1 in the ER. In contrast, APP-C99 did not colocalize with PS1, but was efficiently processed by PS1-dependent gamma-secretase. APP-C99 resides in a compartment that is negative for ER, intermediate compartment, and Golgi marker proteins. We conclude that gamma-secretase cleavage of APP-C99 occurs in a specialized subcellular compartment where little or no PS1 is detected. This suggests that at least one other factor than PS1, located downstream of the ER, is required for the gamma-cleavage of APP-C99. In agreement, we found that intracellular gamma secretase processing of APP-C99-KK both at the gamma40 and the gamma42 site could be restored partially after brefeldin A treatment. Our data confirm the "spatial paradox" and raise several questions regarding the PS1 is gamma-secretase hypothesis. PMID- 11502764 TI - Thylakoid DeltapH-dependent precursor proteins bind to a cpTatC-Hcf106 complex before Tha4-dependent transport. AB - The thylakoid DeltapH-dependent pathway transports folded proteins with twin arginine-containing signal peptides. Identified components of the machinery include cpTatC, Hcf106, and Tha4. The reaction occurs in two steps: precursor binding to the machinery, and transport across the membrane. Here, we show that a cpTatC-Hcf106 complex serves as receptor for specific binding of twin arginine containing precursors. Antibodies to either Hcf106 or cpTatC, but not Tha4, inhibited precursor binding. Blue native gel electrophoresis and coimmunoprecipitation of digitonin-solubilized thylakoids showed that Hcf106 and cpTatC are members of an approximately 700-kD complex that lacks Tha4. Thylakoid bound precursor proteins were also associated with an approximately 700-kD complex and were coimmunoprecipitated with antibodies to cpTatC or Hcf106. Chemical cross-linking revealed that precursors make direct contact with cpTatC and Hcf106 and confirmed that Tha4 is not associated with precursor, cpTatC, or Hcf106 in the membrane. Precursor binding to the cpTatC-Hcf106 complex required both the twin arginine and the hydrophobic core of the signal peptide. Precursors remained bound to the complex when Tha4 was sequestered by antibody, even in the presence of DeltapH. These results indicate that precursor binding to the cpTatC Hcf106 complex constitutes the recognition event for this pathway and that subsequent participation by Tha4 leads to translocation. PMID- 11502765 TI - Clinical review 134: The endocrinology of prostate cancer. PMID- 11502766 TI - Editorial: The rational use of growth hormone in HIV-infected patients. PMID- 11502767 TI - The effects of recombinant human growth hormone on body composition and glucose metabolism in HIV-infected patients with fat accumulation. AB - GH has been proposed as a therapy for patients with HIV-associated fat accumulation, but the pharmacological doses (6 mg/d) used have been associated with impaired fasting glucose and hyperglycemia. In contrast, physiologic doses of GH ( approximately 1 mg/d) in HIV-negative men reduced visceral adiposity and eventually improved insulin sensitivity, despite initially causing insulin resistance. We conducted an open-label study to evaluate the effects of a lower pharmacologic dose of GH (3 mg/d) in eight men with HIV-associated fat accumulation. Oral glucose tolerance, insulin sensitivity, and body composition were measured at baseline, and 1 and 6 months. Six patients completed 1 month and 5, 6 months of GH therapy. IGF-I levels increased 4-fold within 1 month of GH treatment. Over 6 months, GH reduced buffalo hump size and excess visceral adipose tissue. Total body fat decreased (17.9 +/- 10.9 to 13.5 +/- 8.4 kg, P = 0.05), primarily in the trunk region. Lean body mass increased (62.9 +/- 6.4 to 68.3 +/- 9.1 kg, P = 0.03). Insulin-mediated glucose disposal, measured by a euglycemic hyperinsulinemic clamp, declined at month 1 (49.7 +/- 27.5 to 25.6 +/- 6.6 nmol/kg(LBM).min/pmol(INSULIN)/liter, P = 0.04); values improved at month 6 (49.2 +/- 22.6, P = 0.03, compared with month 1) and did not differ significantly from baseline. Similarly, the integrated response to an oral glucose load worsened at month 1 (glucose area under the curve 20.1 +/- 2.3 to 24.6 +/- 3.7 mmol.h/liter, P < 0.01), whereas values improved at month 6 (22.1 +/- 1.5, P = 0.02, compared with month 1) and did not differ significantly from baseline. One patient developed symptomatic hyperglycemia within 2 wk of GH initiation; baseline oral glucose tolerance testing revealed preexisting diabetes despite normal fasting glucose. In conclusion, GH at 3 mg/d resulted in a decrease in total body fat and an increase in lean body mass in this open-label trial. While insulin sensitivity and glucose tolerance initially worsened, they subsequently improved toward baseline. However, the dose of GH used in this trial was supraphysiologic and led to an increase in IGF-I levels up to three times the upper normal range. Because there are known adverse effects of long-term GH excess, the effectiveness of lower doses of GH should be studied. We also recommend a screening oral glucose tolerance test be performed to exclude subjects at risk for GH-induced hyperglycemia. PMID- 11502768 TI - The effect of methimazole pretreatment on the efficacy of radioactive iodine therapy in Graves' hyperthyroidism: one-year follow-up of a prospective, randomized study. AB - The effect of antithyroid drugs on the efficacy of radioiodine (131I) treatment is still controversial. This study evaluated the effect of methimazole pretreatment on the efficacy of 131I therapy in Graves' hyperthyroidism. Sixty one untreated patients were randomly assigned to receive 131I alone (32 patients) or 131I plus pretreatment with methimazole (30 mg/d; 29 patients). 131I was administered 4 d after drug discontinuation. The calculated 131I dose was 200 microCi/g thyroid tissue as estimated by ultrasound, corrected by 24-h radioiodine uptake. Serum TSH, T4, and free T4 were measured 4 d before 131I therapy, on the day of treatment, and then monthly for 1 yr. Considering cure as euthyroidism or hypothyroidism, based on free T4 measurement, approximately 80% of patients from both groups were cured 3 months after beginning 131I treatment. After 1 yr the groups were similar in terms of persistent hyperthyroidism (15.6% vs. 13.8%), euthyroidism (28.1% vs. 31.0%), or hypothyroidism (56.3% vs. 55.2%). Relapsed patients presented larger thyroid volume (P = 0.002), higher 24-h radioiodine uptake (P = 0.022), and T3 levels (P = 0.002). Multiple logistic regression analysis identified T3 values as an independent predictor of therapy failure. In conclusion, pretreatment with methimazole had no effect on either the time required for cure or the 1-yr success rate of 131I therapy. PMID- 11502769 TI - A prospective investigation of quality of life and psychological well-being after the discontinuation of GH treatment in adolescent patients who had GH deficiency during childhood. AB - Some patients given growth-promoting therapy for GH deficiency in childhood will remain GH deficient in their adult lives and hence could benefit from continued GH replacement therapy. This longitudinal study sought to assess whether quality of life declines after GH discontinuation in late adolescence, and whether differences can be discerned in quality of life in patients whose GH deficiency persists into adulthood and those whose GH secretory capacity falls within normal ranges. Forty patients, aged 16-21 yr at baseline, were assessed over a 2-yr period commencing with discontinuation of GH therapy. Twenty-one patients were assigned to a GH deficiency group, and 19 were assigned to a GH-sufficient group. Quality of life assessments were made using the Nottingham Health Profile, Psychological General Well-Being Index, and Mood Adjective Check List Measures. Visual analog assessment of personality and affect and cognitive function tests were performed. The Mood Adjective Check List and visual analog assessments identified between-group and temporal changes in a limited number of the various personality domains assessed. The Psychological General Well-Being Index assessment indicated greater baseline impairment in the GH deficiency group than in the GH-sufficient group in overall score and in the domains of depression and general health. There was also a between-group difference in anxiety score at the 2-yr assessment, with the GH deficiency group having greater anxiety. Measurement of cognitive factors failed to reveal differences between groups. These results indicate that the discontinuation of GH therapy in late adolescence does not risk an immediate decline in the perceived quality of life detectable with the Nottingham Health Profile and Psychological General Well-Being Index measures. However, differences detected with the Mood Adjective Check List and visual analog assessments hint at clinically significant changes in the life experiences of adolescents discontinued from GH for which traditional measures may lack sensitivity. PMID- 11502770 TI - Circadian and ultradian rhythm and leptin pulsatility in adult GH deficiency: effects of GH replacement. AB - Leptin contributes to the regulation of body weight in healthy individuals and is secreted by adipocytes in a diurnal pattern, with superimposed pulsatility. The circulating leptin concentration is increased in both normally obese and untreated adult GH deficiency, a syndrome characterized by increased adiposity. Leptin circadian rhythm is preserved in adult GH deficiency patients; however, an ultradian rhythm and pulsatility has previously not been reported. Alterations in plasma leptin concentration in obese individuals and adult GH deficiency patients after GH replacement have been attributed to changes in body fat mass. In our present study leptin circadian and ultradian rhythm, leptin pulsatility and its relationship with body fat mass were examined in 12 adult GH deficiency patients (6 men) before and 1 month after GH replacement. All subjects with adult GH deficiency had hypopituitarism subsequent to pituitary surgery and were stabilized on conventional pituitary hormone replacement. Plasma leptin was measured over 24 h at 30-min intervals, and changes in body composition were recorded using bioelectrical impedance. The 24-h mean leptin concentration decreased from 2.04 +/- 0.04 nmol/liter in untreated adult GH deficiency patients to 1.64 +/- 0.03 nmol/liter after 1 month of GH replacement (P < 0.0001). Before GH replacement, patients demonstrated a significant mean leptin circadian rhythm (P < 0.001), with a mesor of 2.05 +/- 0.03 nmol/liter and a superimposed ultradian frequency of 2.0 +/- 0.1 cycles/d. After GH replacement, the circadian rhythm was preserved (P < 0.001), but mesor decreased to 1.65 +/- 0.01 nmol/liter (P < 0.0001), and leptin ultradian frequency increased to 16.0 +/-0.2 cycles/d (P < 0.0001). Pulse analysis (ULTRA) revealed 3.1 +/- 0.9 pulses/24 h in untreated adult GH deficiency patients, which significantly increased to 9.9 +/- 2.2 pulses/24 h after 1 month of GH replacement (P < 0.001). There was no significant change in body mass index or body fat mass after 1 month of GH replacement. The body fat percentage significantly reduced from 36.5 +/- 2.8% to 35.5 +/- 2.7% after 1 month of GH replacement (P < 0.05). This change in body fat percentage was explained by a significant increase in lean body mass, from 56.2 +/- 2.8 kg at baseline to 57.4 +/- 2.7 kg after 1 month (P < 0.05). A significant correlation was observed between plasma leptin and body fat percentage at baseline and 1 month after GH replacement (both, r = 0.7; P < 0.01) in the absence of a significant correlation between leptin and body fat mass before and after GH replacement (P = 0.13 and P = 0.11, respectively). Thus, untreated adult GH deficiency is associated with elevated 24-h leptin concentration, preserved circadian rhythm, and decreased pulsatility. The secretory pattern is restored after GH replacement, with a significant reduction in the 24-h mean leptin concentration, maintenance of circadian rhythm, and increased pulsatility. This GH-induced change in the leptin secretory pattern precedes significant changes in body fat mass and may therefore be independent of changes in adipose tissue. Restoration of leptin pulsatility may be of clinical benefit, and our data could lead to novel approaches for leptin manipulation in the future. PMID- 11502771 TI - The nonimpact of thyroid stunning: remnant ablation rates in 131I-scanned and nonscanned individuals. AB - Thyroid stunning has been reported as the temporary impairment of thyroid tissue after a 111-MBq or greater diagnostic 131I dose that decreases the final absorbed dose in ablative therapy. Concerns regarding the reality of stunning have arisen in part due to a flawed study design in prior reports. To assess whether a stunning effect has any impact on therapeutic outcomes, we compared initial treatment ablation rates in patients who received 111- to 185-MBq 131I diagnostic scans (n = 37) before ablative doses of 3700-7400 MBq with ablation rates in patients who did not receive any 131I before the initial treatment dose (n = 63). Ablation rates were 64.9% for scanned patients and 66.7% for nonscanned patients, a nonsignificant difference. Nonscanned patients with metastatic lesions (n = 23) were ablated at a higher rate (78.3%) than scanned patients (n = 9) (66.7%), but the difference was not significant (P = 0.50). It is possible that the reported stunning phenomenon, specifically its impact in temporarily impairing tissue, has been overemphasized. PMID- 11502772 TI - 131I therapy for differentiated thyroid cancer leads to an earlier onset of menopause: results of a retrospective study. AB - Treatment with 131I for differentiated thyroid cancer may give a follicle damaging radiation dose to the ovaries. This damage to the ovarian function could shorten the fertile life span and advance the natural menopause. To address this issue, we studied retrospectively the menopausal age of 130 women treated with 131I for differentiated thyroid cancer in our institution from 1974-1993. The menopausal age of women treated with 131I for differentiated thyroid cancer after total thyroidectomy and subjected to suppressive L-T4 therapy was compared with the menopausal age of a control group including 127 goitrous women who were treated with suppressive L-T4 for a comparable period of time. The cumulative therapeutic 131I dose to cancer patients ranged from 1,110-40,700 MBq (mean +/- SD, 5,308 +/- 5,483 MBq; median, 3700 MBq). All patients chosen for the study were younger than 45 yr when first treated (i.e. first administration of 131I and L-T4 for cancer patients, and institution of L-T4 therapy for goitrous patients), and older than 45 yr at the end of the study period. The menopausal status of both groups was assessed from the clinical records and compared using Kaplan Meier survival analysis. The menopausal age of cancer women treated with 131I and suppressive L-T4 therapy was less than that of goitrous patients treated with suppressive L-T4 therapy (P < 0.001). We could not detect any relationship between menopausal age and the age at the first or last 131I dose or to the cumulative 131I dose received. These data indicate that 131I treatment is probably associated with an earlier ovarian failure in thyroid cancer patients. Conceivably, the ovarian irradiation by 131I might contribute to the process of the follicular atresia, thus inducing earlier menopause. PMID- 11502773 TI - Sexual dimorphism in counterregulatory responses to hypoglycemia after antecedent exercise. AB - After antecedent hypoglycemia, counterregulatory responses to subsequent hypoglycemia exhibit greater blunting in men than in women. Because physical exercise and hypoglycemia share multiple counterregulatory mechanisms, we hypothesized that prior exercise may also result in gender-specific blunting of counterregulatory responses to subsequent hypoglycemia. Thirty healthy subjects (15 women and 15 men; age, 28 +/- 3 yr; body mass index, 23 +/- 1 kg/m2) were studied during 2-d experiments. Day 1 consisted of either identical 90-min morning and afternoon cycle exercise at 50% maximum oxygen expenditure or two 2-h episodes of hyperinsulinemic euglycemia. Day 2 consisted of a 2-h morning hyperinsulinemic-hypoglycemic clamp. Endogenous glucose production was measured using [3-(3)H]glucose. Muscle sympathetic nerve activity was measured using microneurography. Day 2 insulin (540 +/- 36 pmol/liter) and plasma glucose (2.9 +/- 0.06 pmol/liter) levels were similar in men and women during the last 30 min of hypoglycemia. Compared with antecedent euglycemia, d 1 exercise produced significant blunting of d 2 counterregulatory responses to hypoglycemia. Several key d 2 counterregulatory responses were blunted to a greater extent in men than in women: glucagon (men, -105 +/- 14; women, -25 +/- 7 ng/liter; P < 0.0001), epinephrine (men, -2625 +/- 257 pmol/liter; women, -212 +/- 573; P < 0.001), norepinephrine (men, -0.50 +/- 0.12 nmol/liter; women, -0 +/- 0.11; P < 0.001), and muscle sympathetic nerve activity (men, -13 +/- 4; women, -4 +/- 4 bursts/min; P < 0.01). Cardiovascular responses (heart rate and systolic and mean arterial blood pressures) were also more blunted by antecedent exercise in men than in women. After d 1 exercise, the amount of glucose infused during d 2 hypoglycemia in men was increased 6-fold compared with that after d 1 euglycemia. This amount was significantly increased (P < 0.01) compared with the 2-fold (P < 0.01) increment in glucose infusion that was required in women after d 1 exercise. Lipolysis was unaffected by d 1 exercise in women, but was significantly blunted during d 2 hypoglycemia in men. In summary, two bouts of prolonged, moderate exercise (90 min at 50% maximum oxygen expenditure) induced a marked sexual dimorphism in key neuroendocrine (glucagon, catecholamines, and muscle sympathetic nerve activity) and metabolic (glucose kinetic, lipolysis) responses to next day hypoglycemia. PMID- 11502774 TI - Thyroid function and autoimmunity during interferon beta-1b treatment: a multicenter prospective study. AB - Thyroid dysfunction and autoimmunity have been reported during type I interferon therapy, namely interferon-alpha for chronic hepatitis or interferon-beta for multiple sclerosis. To define the frequency of thyroid dysfunction and autoimmunity during interferon-beta treatment, 156 multiple sclerosis patients were prospectively followed up by 18 centers for 1 yr after starting interferon beta-1b treatment. Serial clinical assessments and tests of thyroid and liver function and antithyroid autoantibodies (all performed by the same centralized laboratory) were conducted every 3 months. TSH and antithyroid autoantibodies against human TG or thyroid microsomal antigens were measured by immunoradiometric methods; free T3 and T4 were measured by chromatographic assays. Longitudinal occurrence of thyroid or liver alterations or of autoantibodies was analyzed with the generalized estimating equations method, correcting for the correlation of repeated measurements of the same subject over time. Pretreatment comparison with a control group of 437 healthy blood donors did not show significant differences in the frequency of thyroid dysfunction or antithyroid autoantibody positivity. During interferon-beta treatment, the de novo frequency of thyroid alteration was 8.3%, that of liver alteration was 37.5%, and that of antithyroid autoantibody was 4.5%. Generalized estimating equations analysis demonstrated that the frequency of liver alteration significantly increased during treatment compared with the baseline value (odds ratio, 7.03; confidence interval, 2.49-19.9), whereas that of thyroid alteration or of antithyroid autoantibodies did not. The frequency of thyroid dysfunction during interferon-beta treatment showed random, nonsignificant changes over time and, in addition, was not correlated to antithyroid autoantibody positivity. PMID- 11502775 TI - Reduced bone density in androgen-deficient women with acquired immune deficiency syndrome wasting. AB - Women with acquired immune deficiency syndrome wasting are at an increased risk of osteopenia because of low weight, changes in body composition, and hormonal alterations. Although women comprise an increasing proportion of human immunodeficiency virus-infected patients, prior studies have not investigated bone loss in this expanding population of patients. In this study we investigated bone density, bone turnover, and hormonal parameters in 28 women with acquired immune deficiency syndrome wasting and relative androgen deficiency (defined as free testosterone < or =3.0 pg/ml, weight < or =90% ideal body weight, weight loss > or =10% from preillness maximum weight, or weight <100% ideal body weight with weight loss > or =5% from preillness maximum weight). Total body (1.04 +/- 0.08 vs. 1.10 +/- 0.07 g/cm2, human immunodeficiency virus-infected vs. control respectively; P < 0.01), anteroposterior lumbar spine (0.94 +/- 0.12 vs. 1.03 +/- 0.09 g/cm2; P = 0.005), lateral lumbar spine (0.71 +/- 0.14 vs. 0.79 +/- 0.09 g/cm2; P = 0.02), and hip (Ward's triangle; 0.68 +/- 0.14 vs. 0.76 +/- 0.12 g/cm2; P = 0.05) bone density were reduced in the human immunodeficiency virus infected compared with control subjects. Serum N-telopeptide, a measure of bone resorption, was increased in human immunodeficiency virus-infected patients, compared with control subjects (14.6 +/- 5.8 vs. 11.3 +/- 3.8 nmol/liter bone collagen equivalents, human immunodeficiency virus-infected vs. control respectively; P = 0.03). Although body mass index was similar between the groups, muscle mass was significantly reduced in the human immunodeficiency virus infected vs. control subjects (16 +/- 4 vs. 21 +/- 4 kg, human immunodeficiency virus-infected vs. control, respectively; P < 0.0001). In univariate regression analysis, muscle mass (r = 0.53; P = 0.004) and estrogen (r = 0.51; P = 0.008), but not free testosterone (r = -0.05, P = 0.81), were strongly associated with lumbar spine bone density in the human immunodeficiency virus-infected patients. The association between muscle mass and bone density remained significant, controlling for body mass index, hormonal status, and age (P = 0.048) in multivariate regression analysis. These data indicate that both hormonal and body composition factors contribute to reduced bone density in women with acquired immune deficiency syndrome wasting. Anabolic strategies to increase muscle mass may be useful to increase bone density among osteopenic women with acquired immune deficiency syndrome wasting. PMID- 11502776 TI - Remission and recurrence of hyperthyroid Graves' disease during and after methimazole treatment when assessed by IgE and interleukin 13. AB - We analyzed the relationship between serum IgE concentrations and the remission or recurrence of Graves' disease. One hundred seven patients with Graves' disease were treated with methimazole (MMI). Serum IgE concentration greater than 170 IU/ml was found in 41 of 107 untreated patients (38.3%). However, the presence of TSH-binding inhibiting immunoglobulin or thyroid-stimulating antibody did not correlate with the IgE concentrations. Remission was found in 20 of 41 patients with elevated IgE concentrations (48.8%) after 18 months of MMI treatment, as opposed to 53 of 66 patients with normal concentrations (80.3%) (P = 0.0014). MMI treatment was discontinued in 73 patients who were followed for 26-48 months. The recurrence of Graves' disease was found in 13 patients, whereas the remaining 60 were still in remission. The rate of long-standing remission was lower in patients with elevated than normal IgE concentration (34.1% vs. 69.7%, P = 0.0007). We also analyzed serum levels of interleukin (IL)-13. Although IL-13 was not detected in all patients, the detection rate was higher in patients without remission and in those with recurrence than in those with long-standing remission (47.1%, 38.5%, and 13.3%, respectively; P = 0.0012). More patients with elevated IgE were positive for allergic diseases and for family history of allergic diseases in their first-degree relatives. We conclude that the elevation of IgE and the higher detection rate of IL-13 are associated with both remission and recurrence of Graves' disease. PMID- 11502777 TI - Hypothalamo-pituitary-adrenal axis dysfunction in chronic fatigue syndrome, and the effects of low-dose hydrocortisone therapy. AB - These neuroendocrine studies were part of a series of studies testing the hypotheses that 1) there may be reduced activity of the hypothalamic-pituitary adrenal axis in chronic fatigue syndrome and 2) low-dose augmentation with hydrocortisone therapy would improve the core symptoms. We measured ACTH and cortisol responses to human CRH, the insulin stress test, and D-fenfluramine in 37 medication-free patients with CDC-defined chronic fatigue syndrome but no comorbid psychiatric disorders and 28 healthy controls. We also measured 24-h urinary free cortisol in both groups. All patients (n = 37) had a pituitary challenge test (human CRH) and a hypothalamic challenge test [either the insulin stress test (n = 16) or D-fenfluramine (n = 21)]. Baseline cortisol concentrations were significantly raised in the chronic fatigue syndrome group for the human CRH test only. Baseline ACTH concentrations did not differ between groups for any test. ACTH responses to human CRH, the insulin stress test, and D- fenfluramine were similar for patient and control groups. Cortisol responses to the insulin stress test did not differ between groups, but there was a trend for cortisol responses both to human CRH and D-fenfluramine to be lower in the chronic fatigue syndrome group. These differences were significant when ACTH responses were controlled. Urinary free cortisol levels were lower in the chronic fatigue syndrome group compared with the healthy group. These results indicate that ACTH responses to pituitary and hypothalamic challenges are intact in chronic fatigue syndrome and do not support previous findings of reduced central responses in hypothalamic-pituitary-adrenal axis function or the hypothesis of abnormal CRH secretion in chronic fatigue syndrome. These data further suggest that the hypocortisolism found in chronic fatigue syndrome may be secondary to reduced adrenal gland output. Thirty-two patients were treated with a low-dose hydrocortisone regime in a double-blind, placebo-controlled cross-over design, with 28 days on each treatment. They underwent repeated 24-h urinary free cortisol collections, a human CRH test, and an insulin stress test after both active and placebo arms of treatment. Looking at all subjects, 24-h urinary free cortisol was higher after active compared with placebo treatments, but 0900-h cortisol levels and the ACTH and cortisol responses to human CRH and the insulin stress test did not differ. However, a differential effect was seen in those patients who responded to active treatment (defined as a reduction in fatigue score to the median population level or less). In this group, there was a significant increase in the cortisol response to human CRH, which reversed the previously observed blunted responses seen in these patients. We conclude that the improvement in fatigue seen in some patients with chronic fatigue syndrome during hydrocortisone treatment is accompanied by a reversal of the blunted cortisol responses to human CRH. PMID- 11502778 TI - Relationship of serum sex steroid levels to longitudinal changes in bone density in young versus elderly men. AB - Estrogen appears to play an important role in determining bone mineral density in men, but it remains unclear whether estrogen primarily determines peak bone mass or also affects bone loss in elderly men. Thus, we assessed longitudinal rates of change in bone mineral density in young (22-39 yr; n = 88) vs. elderly (60-90 yr; n = 130) men and related these to circulating total and bioavailable estrogen and testosterone levels. In young men bone mineral density increased significantly over 4 yr at the mid-radius and ulna and at the total hip (by 0.32-0.43%/yr), whereas it decreased in the elderly men at the forearm sites (by 0.49-0.66%/yr), but did not change at the total hip. The rate of increase in bone mineral density at the forearm sites in the young men was significantly correlated to serum total and bioavailable estradiol and estrone levels (r = 0.22-0.35), but not with total or bioavailable testosterone levels. In the elderly men the rates of bone loss at the forearm sites were most closely associated with serum bioavailable estradiol levels (r = 0.29-0.33) rather than bioavailable testosterone levels. Moreover, elderly men with bioavailable estradiol levels below the median [40 pmol/liter (11 pg/ml)] had significantly higher rates of bone loss and levels of bone resorption markers than men with bioavailable estradiol levels above 40 pmol/liter. These data thus indicate that estrogen plays a key role both in the acquisition of peak bone mass in young men and in bone loss in elderly men. Moreover, our findings suggest that age-related decreases in bioavailable estradiol levels to below 40 pmol/liter may well be the major cause of bone loss in elderly men. This subset of men is perhaps most likely to benefit from preventive therapy. PMID- 11502779 TI - Comparison of the effectiveness and tolerability of intravenous or oral glucocorticoids associated with orbital radiotherapy in the management of severe Graves' ophthalmopathy: results of a prospective, single-blind, randomized study. AB - Eighty-two consecutive patients with moderate-to-severe and active Graves' ophthalmopathy were randomly treated with orbital radiotherapy combined with either oral (prednisone; starting dose, 100 mg/d; withdrawal after 5 months) or iv (methylprednisolone; 15 mg/kg for four cycles and then 7.5 mg/kg for four cycles; each cycle consisted of two infusions on alternate days at 2-wk intervals) glucocorticoids. The two groups did not differ for age, gender, duration of hyperthyroidism and ophthalmopathy, prevalence of smokers, thyroid volume, and pretreatment ocular conditions. Both groups of patients received radioiodine therapy shortly before treatment for Graves' ophthalmopathy. Follow up lasted for 12 months. A significant reduction in proptosis (from 23.2 +/- 3.0 to 21.6 +/- 1.2 mm in the iv glucocorticoid group, P < 0.0001; and from 23 +/- 1.8 to 21.7 +/- 1.8 mm in oral glucocorticoid group, P < 0.0001) and in lid width (from 13.3 +/- 2.5 to 11.8 +/- 2.2 mm, and from 13.6 +/- 2.0 to 11.5 +/- 1.9 mm, respectively; P < 0.001 in both cases) occurred, with no difference between the two groups. Diplopia significantly improved in both groups: it disappeared in 13 of 27 (48.1%) iv glucocorticoid patients (P < 0.005) and in 12 of 33 (36.4%) oral glucocorticoid patients (P < 0.03). The degree of amelioration of diplopia did not significantly differ between the two groups (P = 0.82). Optic neuropathy improved in 11 of 14 iv glucocorticoid (P < 0.01) and only in 3 of 9 oral glucocorticoid (P = 0.57) patients, with no significant difference in these outcomes. The Clinical Activity Score decreased from 4.5 +/- 1.2 to 1.7 +/- 1.0 (P < 0.0001) in the iv glucocorticoid group and from 4.2 +/- 1.1 to 2.2 +/- 1.2 (P < 0.0001) in the oral glucocorticoid group; final Clinical Activity Score was significantly lower in iv glucocorticoid than in oral glucocorticoid patients (P < 0.01). By self-assessment evaluation, 35 (85.3%) iv glucocorticoid and 30 (73.2%) oral glucocorticoid patients reported an improvement of ocular conditions (P = 0.27). Overall, both treatments produced favorable effects in most patients, but responders in the iv glucocorticoid group (36 of 41, 87.8%) were more than in the oral glucocorticoid group (26 of 41, 63.4%) (P < 0.02). Moreover, iv glucocorticoid treatment was better tolerated than oral glucocorticoid treatment. Side effects occurred in 23 (56.1%) iv glucocorticoid and 35 (85.4%) oral glucocorticoid patients (P < 0.01); in particular, cushingoid features developed in 5 of the former and 35 of the latter patients. One iv glucocorticoid patient had severe hepatitis of undetermined origin at the end of glucocorticoid treatment, followed by spontaneous recovery. In conclusion, high-dose iv glucocorticoid and oral glucocorticoid (associated with orbital radiotherapy) are effective in the management of severe Graves' ophthalmopathy, but the iv route seems to be more effective and better tolerated than the oral route and associated with a lower rate of side effects. PMID- 11502780 TI - Successful long-term treatment of refractory Cushing's disease with high-dose mifepristone (RU 486). AB - An extremely ill patient, with Cushing's syndrome caused by an ACTH-secreting pituitary macroadenoma, experienced complications of end-stage cardiomyopathy, profound psychosis, and multiple metabolic disturbances. Initially treated unsuccessfully by a combination of conventional surgical, medical, and radiotherapeutic approaches, he responded dramatically to high-dose long-term mifepristone therapy (up to 25 mg/kg x d). Treatment efficacy was confirmed by the normalization of all biochemical glucocorticoid-sensitive measurements, as well as by the significant reversal of the patient's heart failure, the resolution of his psychotic depression, and the eventual unusual return of his adrenal axis to normal. His 18-month-long mifepristone treatment course was notable for development of severe hypokalemia that was attributed to excessive cortisol activation of the mineralocorticoid receptor, which responded to spironolactone administration. This case illustrates the efficacy of high-dose long-term treatment with mifepristone in refractory Cushing's syndrome. The case also demonstrates the potential need for concomitant mineralocorticoid receptor blockade in mifepristone-treated Cushing's disease, because cortisol levels may rise markedly, reflecting corticotroph disinhibition, to cause manifestations of mineralocorticoid excess. PMID- 11502781 TI - Insulin resistance as a predictor of age-related diseases. AB - The current study was initiated to evaluate the ability of insulin resistance to predict a variety of age-related diseases. Baseline measurements of insulin resistance and related variables were made between 1988-1995 in 208 apparently healthy, nonobese (body mass index < 30 kg/m2) individuals, who were then evaluated 4-11 yr later (mean +/- SEM = 6.3 +/- 0.2 yr) for the appearance of the following age-related diseases: hypertension, coronary heart disease, stroke, cancer, and type 2 diabetes. The effect of insulin resistance on the development of clinical events was evaluated by dividing the study group into tertiles of insulin resistance at baseline and comparing the events in these 3 groups. Clinical endpoints (n = 40) were identified in 37 individuals (18%) of those evaluated, including 12 with hypertension, 3 with hypertension + type 2 diabetes, 9 with cancer, 7 with coronary heart disease, 4 with stroke, and 2 with type 2 diabetes. Twenty-eight out of the total 40 clinical events were seen in 25 individuals (36%) in the most insulin-resistant tertile, with the other 12 occurring in the group with an intermediate degree of insulin resistance. Furthermore, insulin resistance was an independent predictor of all clinical events, using both multiple logistic regression and Cox's proportional hazards analysis. The fact that an age-related clinical event developed in approximately 1 out of 3 healthy individuals in the upper tertile of insulin resistance at baseline, followed for an average of 6 yr, whereas no clinical events were observed in the most insulin-sensitive tertile, should serve as a strong stimulus to further efforts to define the role of insulin resistance in the genesis of age related diseases. PMID- 11502783 TI - Sensitization to insulin induces ovulation in nonobese adolescents with anovulatory hyperandrogenism. AB - In nonobese girls with an adolescent variant of polycystic ovary syndrome, insulin-sensitizing treatment reduces hyperinsulinism, dyslipidemia, and hyperandrogenism and restores eumenorrhea; however, the effect on anovulation is unknown. We assessed whether metformin treatment is capable of inducing ovulation in nonobese adolescents with anovulatory hyperandrogenism after precocious pubarche. The study population consisted of 18 adolescents (mean age, 16 yr; body mass index, 21.4 kg/m2; 3-7 yr beyond menarche) with hyperinsulinemic hyperandrogenism. All girls received metformin for 6 months in a daily dose of 1275 mg. Before inclusion, persistent anovulation was documented by weekly serum progesterone measurements less than 4 ng/ml (months -3 and -1); the ovulation rate was assessed similarly after 2, 4 and 6 months on metformin; a premenstrual progesterone level greater than 8 ng/ml was used as ovulation marker. Regular menses were reported by 16 of 18 girls within 4 months on metformin, and all girls were eumenorrheic after 6 months on metformin. Of the 18 girls, 1 (6%) ovulated after 2 months on metformin, 7 (39%) after 4 months, and 14 (78%) after 6 months; ovulation induction failed in the girls with the lowest birth weight or most severe hyperandrogenism. Metformin treatment was well tolerated. In conclusion, sensitization to insulin was found to be an effective approach to induce ovulation in nonobese adolescents with anovulatory hyperandrogenism. PMID- 11502782 TI - Usefulness of L-carnitine, a naturally occurring peripheral antagonist of thyroid hormone action, in iatrogenic hyperthyroidism: a randomized, double-blind, placebo-controlled clinical trial. AB - Old studies in animals and unblinded studies in a few hyperthyroid patients suggested that L -carnitine is a periferal antagonist of thyroid hormone action at least in some tissues. This conclusion was substantiated by our recent observation that carnitine inhibits thyroid hormone entry into the nucleus of hepatocytes, neurons, and fibroblasts. In the randomized, double-blind, placebo controlled 6-month trial reported here, we assessed whether 2 or 4 g/d oral L carnitine were able to both reverse and prevent/minimize nine hyperthyroidism- related symptoms. We also evaluated changes on nine thyroid hormone-sensitive biochemical parameters and on vertebral and hip mineral density (bone mineral density). Fifty women under a fixed TSH-suppressive dose of L -T(4) for all 6 months were randomly allocated to five groups of 10 subjects each. Group 0 associated placebo for 6 months; groups A2 and A4 started associating placebo (first bimester), substituted placebo with 2 or 4 g/d carnitine (second bimester), and then returned to the association with placebo. Groups B2 and B4 started associating 2 and 4 g/d carnitine for the first two bimesters, and then substituted carnitine with placebo (third bimester). Symptoms and biochemical parameters worsened in group 0. In group A, symptoms and biochemical parameters worsened during the first bimester, returned to baseline or increased minimally during the second bimester (except osteocalcin and urinary OH-proline), and worsened again in the third bimester. In group B, symptoms and biochemical parameters (except osteocalcin and urinary OH-proline) did not worsen or even improved over the first 4 months; they tended to worsen in the third bimester. In both the A and B groups, the two doses of carnitine were similarly effective. At the end of the trial, bone mineral density tended to increase in groups B and A (B > A). In conclusion, L-carnitine is effective in both reversing and preventing symptoms of hyperthyroidism and has a beneficial effect on bone mineralization. Because hyperthyroidism depletes the body deposits of carnitine and since carnitine has no toxicity, teratogenicity, contraindications and interactions with drugs, carnitine can be of clinical use. PMID- 11502784 TI - Serum Tg--a sensitive marker of thyroid abnormalities and iodine deficiency in epidemiological studies. AB - Serum Tg is widely used in the control of thyroid cancer but also in the diagnosis of certain other thyroid diseases. Serum Tg may be useful in the characterization of the iodine status of a population, but little is known about determinants of serum Tg levels. We examined a random selection of 4,649 subjects from 2 regions in Denmark with different iodine status. Thyroid volume and structure were determined with ultrasonography, and thyroid function tests and Tg analysis were performed. The factor with the closest association with serum Tg levels was thyroid volume at ultrasonography (P < 0.001). Also thyroid nodularity (P < 0.001) and iodine excretion (P < 0.001) had close associations to serum Tg, even after adjusting for the influence of the other parameters. Thyroid dysfunction had a less pronounced but still highly significant association with serum Tg (P < 0.001), but no relation was found to serum TSH in general. The association with age seemed to rely on differences in the prevalence of thyroid abnormalities, and men had lower Tg levels than women of the same age. There was a marked difference in serum Tg between the two regions with slightly different iodine excretion also after adjusting for the other factors. In conclusion, serum Tg reflects thyroid abnormalities and thyroid function and is a sensitive marker of iodine deficiency in a population. PMID- 11502785 TI - Effects of GH and/or sex steroid administration on abdominal subcutaneous and visceral fat in healthy aged women and men. AB - Aging is associated with reduced GH, IGF-I, and sex steroid axis activity and with increased abdominal fat. We employed a randomized, double-masked, placebo controlled, noncross-over design to study the effects of 6 months of administration of GH alone (20 microg/kg BW), sex hormone alone (hormone replacement therapy in women, testosterone enanthate in men), or GH + sex hormone on total abdominal area, abdominal sc fat, and visceral fat in 110 healthy women (n = 46) and men (n = 64), 65-88 yr old (mean, 72 yr). GH administration increased IGF-I levels in women (P = 0.05) and men (P = 0.0001), with the increment in IGF-I levels being higher in men (P = 0.05). Sex steroid administration increased levels of estrogen and testosterone in women and men, respectively (P = 0.05). In women, neither GH, hormone replacement therapy, nor GH + hormone replacement therapy altered total abdominal area, sc fat, or visceral fat significantly. In contrast, in men, administration of GH and GH + testosterone enanthate decreased total abdominal area by 3.9% and 3.8%, respectively, within group and vs. placebo (P = 0.05). Within-group comparisons revealed that sc fat decreased by 10% (P = 0.01) after GH, and by 14% (P = 0.0005) after GH + testosterone enanthate. Compared with placebo, sc fat decreased by 14% (P = 0.05) after GH, by 7% (P = 0.05) after testosterone enanthate, and by 16% (P = 0.0005) after GH + testosterone enanthate. Compared with placebo, visceral fat did not decrease significantly after administration of GH, testosterone enanthate, or GH + testosterone enanthate. These data suggest that in healthy older individuals, GH and/or sex hormone administration elicits a sexually dimorphic response on sc abdominal fat. The generally proportionate reductions we observed in sc and visceral fat, after 6 months of GH administration in healthy aged men, contrast with the disproportionate reduction of visceral fat reported after a similar period of GH treatment of nonelderly GH deficient men and women. Whether longer term administration of GH or testosterone enanthate, alone or in combination, will reduce abdominal fat distribution related cardiovascular risk in healthy older men remains to be elucidated. PMID- 11502786 TI - Radioiodine treatment of hyperthyroidism-prognostic factors for outcome. AB - There is little consensus regarding the most appropriate dose regimen for radioiodine (131I) in the treatment of hyperthyroidism. We audited 813 consecutive hyperthyroid patients treated with radioiodine to compare the efficacy of 2 fixed-dose regimens used within our center (185 megabequerels, 370 megabequerels) and to explore factors that may predict outcome. Patients were categorized into 3 diagnostic groups: Graves' disease, toxic nodular goiter, and hyperthyroidism of indeterminate etiology. Cure after a single dose of 131I was investigated and defined as euthyroid off all treatment for 6 months or T4 replacement for biochemical hypothyroidism in all groups. As expected, patients given a single dose of 370 megabequerels had a higher cure rate than those given 185 megabequerels, (84.6% vs. 66.6%, P < 0.0001) but an increase in hypothyroidism incidence at 1 yr (60.8% vs. 41.3%, P < 0.0001). There was no difference in cure rate between the groups with Graves' disease and those with toxic nodular goiter (69.5% vs. 71.4%; P, not significant), but Graves' patients had a higher incidence of hypothyroidism (54.5% vs. 31.7%, P < 0.0001). Males had a lower cure rate than females (67.6% vs. 76.7%, P = 0.02), whereas younger patients (<40 yr) had a lower cure rate than patients over 40 yr old (68.9% vs. 79.3%, P < 0.001). Patients with more severe hyperthyroidism (P < 0.0001) and with goiters of medium or large size (P < 0.0001) were less likely to be cured after a single dose of 131I. The use of antithyroid drugs, during a period 2 wk before or after 131I, resulted in a significant reduction in cure rate in patients given 185 megabequerels 131I (P < 0.01) but not 370 megabequerels. Logistic regression analysis showed dose, gender, goiters of medium or large size, and severity of hyperthyroidism to be significant independent prognostic factors for cure after a single dose of 131I. We have demonstrated that a single fixed dose of 370 megabequerels 131I is highly effective in curing toxic nodular hyperthyroidism as well as Graves' hyperthyroidism. Because male patients and those with more severe hyperthyroidism and medium or large-sized goiters are less likely to respond to a single dose of radioiodine, we suggest that the value of higher fixed initial doses of radioiodine should be evaluated in these patient categories with lower cure rates. PMID- 11502787 TI - Combination treatment with estrogen and calcitriol in the prevention of age related bone loss. AB - Estrogen deficiency and declining calcium absorption due to reduced calcitriol levels or intestinal resistance to calcitriol, are important factors in the pathogenesis of age-related bone loss. The main objective of this study was to examine the effect of estrogen and 1,25-dihydroxyvitamin D therapy given individually or in combination on bone loss in elderly women. Four hundred eighty nine elderly women with normal bone density for their age, aged 65-77 yr, were entered into a randomized double blind, placebo-controlled trial. Women were randomized to one of four groups: conjugated estrogens (0.625 mg, daily) to women without a uterus (estrogen replacement therapy) plus medroxyprogesterone acetate (2.5 mg, daily) to women with a uterus (hormone replacement therapy), calcitriol (0.25 microg twice daily), a combination of hormone replacement therapy/estrogen replacement therapy plus calcitriol, or placebos for 3 yr. The primary outcome was the change in bone mineral density of the femoral neck and spine. In the intent to treat analysis, hormone therapy (hormone replacement therapy/estrogen replacement therapy) produced a mean (+/-1 SD) increase in bone mineral density of 2.98% (+/-5.45%) at the femoral neck (P < 0.0001) and 4.36% (+/-6.42%) at the spine (P < 0.0001). There were parallel increases in total hip and trochanter bone mineral density. Calcitriol increased bone mineral density 0.10% (+/- 4.27%) at the femoral neck (P = 0.57) and 1.65% (+/- 4.83%) at the spine (P < 0.0124). The combination of hormone replacement therapy/estrogen replacement therapy + calcitriol increased bone mineral density 3.80% (+/-4.95%) at the femoral neck (P < 0.001), 4.91% (+/-6.0%) at the spine (P < 0.0001), and parallel changes at the total hip and trochanter. All three treatment groups differed significantly from placebo at the spine and for the hormone replacement therapy/estrogen replacement therapy groups at the femoral neck, spine, total hip and trochanter. There were no significant differences between combination therapy and hormone replacement therapy/estrogen replacement therapy alone on bone mineral density at any site in the intent to treat analysis. In a secondary analysis of the effect in women who were adherent to treatment, calcitriol had a more significant effect on spine (P = 0.003) and total hip (P = 0.004). The increase in bone mineral density in the adherent groups of women was always higher compared with the intent to treat groups. Combination therapy compared with hormone replacement therapy/estrogen replacement therapy alone produced a significantly greater response in trochanter (P = 0.007) and total hip bone mineral density (P = 0.0017). In summary, hormone replacement therapy/estrogen replacement therapy alone and in combination with calcitriol therapy was highly effective in reducing bone resorption and increasing bone mineral density at the hip and other clinically relevant sites in a group of elderly women, with normal bone density for their age. Calcitriol was effective in increasing spine bone mineral density. In the adherent women, combination therapy with hormone replacement therapy/estrogen replacement therapy and calcitriol increased bone mineral density significantly more in the total hip and trochanter than did hormone replacement therapy/estrogen replacement therapy alone. PMID- 11502788 TI - Prospective randomized study of effects of unopposed estrogen replacement therapy on markers of coagulation and inflammation in postmenopausal women. AB - Estrogen replacement therapy decreases the risk of arterial disease while at the same time increases the risk for venous thrombosis. Whether a common mechanism(s) of coagulation and inflammation contributes to both responses is unclear. This study determined simultaneous effects of estrogen replacement therapy on regulators of the direct (extrinsic) pathway for activation of coagulation, coagulation, and the acute phase response. Plasma from 26 postmenopausal women without risk factors for cardiovascular disease was collected before (baseline) and after 3 months of treatment with either conjugated equine estrogen (Premarin, 0.625 mg/d) or placebo. Plasma lipids, tissue factor pathway inhibitor antigen and activity, plasminogen, prothrombin, P-selectin, alpha1-protease inhibitor, and C-reactive protein were measured. Estrogen replacement therapy significantly reduced mean concentrations of tissue factor pathway inhibitor (antigen and activity; P < 0.001), which were correlated significantly to decreases in low density lipoprotein (r2 = 0.71). Plasminogen and C-reactive protein increased significantly. Other parameters were unchanged. The results of this prospective study suggest that 3 months of estrogen replacement therapy in healthy postmenopausal women decreases low density lipoprotein with simultaneous decreases in tissue factor pathway inhibitor, a major inhibitor of the extrinsic coagulation pathway, and increases C-reactive protein, a component of the acute phase response. Concomitant changes in these parameters may reduce the risk for arterial disease while altering the threshold for thrombotic events. PMID- 11502789 TI - Ovarian hyperstimulation without elevated serum estradiol associated with pure follicle-stimulating hormone-secreting pituitary adenoma. AB - We report a unique case of a 28-yr-old woman with a gonadotroph adenoma secreting FSH, presented with ovarian hyperstimulation, without elevation of serum estradiol. She presented with abdominal pain and large ovaries (both 10 cm in diameter) with multiple follicular cysts shortly after discontinuing oral contraceptive pills. She had a supranormal PRL level of 71 microg/liter (normal, <20), FSH of 8.4-9.2 IU/liter (normal for follicular phase, 2.4-10), LH of 0.01 IU/liter (normal, 1.6-9.3), estradiol of 108 pmol/liter (normal for follicular phase, 80-790), and free alpha-subunit level of 0.11 microg/liter (normal, <1.8). A nuclear magnetic resonance study revealed invasive pituitary macroadenoma, 30 mm in diameter. Dopamine agonist (cabergoline) treatment normalized serum PRL but had no affect on FSH levels. A transsphenoidal surgery was performed, and most of the adenoma was resected. One month after surgery the patient resumed menstruation, and the hormonal profile included serum FSH of 6.3 IU/liter, LH of 2.1 IU/liter, estradiol of 156 pmol/liter, and PRL of 10 microg/liter. The excised adenoma tissue exhibited intense immunostaining for FSH and secreted this hormone to culture medium. Stimulation with TRH (both in vivo preoperatively and in vitro study of the excised tumor) had no effect on FSH secretion from the adenoma. Estradiol did not suppress FSH release from cultured adenoma cells. Patient serum samples showed significant FSH bioactivity when tested in a human granulosa cell line. This case is remarkable because the ovarian hyperstimulation related to the FSH-secreting adenoma was not associated with high levels of serum estradiol, probably due to insufficient LH production by the normal pituitary. Thus, it supports the two-cell, two-gonadotropin theory, that both FSH and LH are necessary for normal ovarian estrogen production. PMID- 11502790 TI - A "pheo" lurks: novel approaches for locating occult pheochromocytoma. AB - Most, but not all, pheochromocytomas can be localized by computed tomography or magnetic resonance imaging. Here we introduce two novel approaches for localization of pheochromocytoma in a patient in whom conventional imaging modalities failed to show the tumor. First, we establish that measurements of plasma free metanephrines coupled with vena caval sampling are useful for localizing occult pheochromocytoma, particularly when elevations in plasma catecholamines are slight or intermittent. Second, we show that positron emission tomographic scanning using the imaging agent 6-[18F]fluorodopamine as a substrate for the norepinephrine transporter offers a highly effective method for tumor localization. These novel approaches may be of value in difficult cases, where biochemical and clinical evidence of pheochromocytoma is compelling, yet conventional imaging modalities fail to locate the tumor. PMID- 11502791 TI - Epitope mapping of tsh receptor-blocking antibodies in Graves' disease that appear during pregnancy. AB - Spontaneous remission of Graves' disease during pregnancy is thought to be due to a reduction of thyroid-stimulating antibody activity. We suspected, however, that a broader change in TSH receptor antibody characteristics might play an important role in modulating disease activity during pregnancy. We measured TSH binding inhibitory Ig, thyroid-stimulating antibody, and thyroid stimulating-blocking antibody activities in 13 pregnant Graves' disease patients at first, second, and third trimesters and 4 months postpartum. To measure and epitope-map thyroid stimulating antibody and thyroid stimulating-blocking antibody activities, we used CHO cells transfected with wild-type human TSH receptor or with several TSH receptor-LH/hCG receptor chimeras: Mc1+2, Mc2, and Mc4. These chimeric cells have their respective TSH receptor residues 9-165, 90-165, and 261-370 substituted with equivalent residues of the LH/hCG receptor. Overall thyroid-stimulating antibody decreased, whereas thyroid stimulating-blocking antibody increased progressively during pregnancy. TSH binding inhibitory Ig fluctuated in individual patients, but overall the activities remained statistically unchanged. Thyroid stimulating-blocking antibody appeared in subjects who were either negative for thyroid-stimulating antibody or whose thyroid-stimulating antibody activity increased or decreased during pregnancy. Epitope mapping showed that the thyroid-stimulating antibodies were mainly directed against residues 9-165 of the N-terminus of the TSH receptor extracellular domain. All thyroid stimulating blocking antibodies had blocking activities against residues 261-370 of the C terminus of the ectodomain. However, the majority of the thyroid stimulating blocking antibodies had a hybrid conformational epitope directed against N terminal residues 9-89 or 90-165 as well. Despite a change in the activity level, we did not observe any change in the epitope of either the stimulatory or blocking Abs as pregnancy advanced. In conclusion, a change in the specificity of TSH receptor antibody from stimulatory to blocking activity was observed during pregnancy, and the appearance of thyroid stimulating-blocking antibody may contribute to the remission of Graves' disease during pregnancy. PMID- 11502792 TI - Metabolism of orally administered androstenedione in young men. AB - Androstenedione is a steroid hormone and the major precursor to testosterone. It is available without prescription and taken with the expectation that it will be converted to testosterone endogenously and increase strength and athletic performance. The metabolism of orally administered testosterone has not been well studied. We randomly assigned 37 healthy men to receive 0, 100, or 300 mg oral androstenedione in a single daily dose for 7 d. Single 8-h urine collections were performed on the day before the start of the androstenedione administration and on d 1 and 7 to assess excretion rates of free and glucuronide- conjugated testosterone, androsterone, etiocholanolone, and dihydrotestosterone. Serum testosterone glucuronide concentrations were measured by frequent blood sampling over 8 h on d 1 in 16 subjects (5 each in the 0 and 100 mg group and 6 in the 300 mg group). In the control group, mean (+/-SE) d 1 and 7 excretion rates for testosterone, androsterone, etiocholanolone, and dihydrotestosterone were 3 +/- 1, 215 +/- 26, 175 +/- 26, and 0.4 +/- 0.1 microg/h, respectively. In the 100 mg group, mean d 1 and 7 excretion rates for testosterone, androsterone, etiocholanolone, and dihydrotestosterone were 47 +/- 11, 3,836 +/- 458, 4,306 +/- 458, and 1.6 +/- 0.2 microg/h, respectively. In the 300 mg group, mean d 1 and 7 excretion rates for testosterone, androsterone, etiocholanolone, and dihydrotestosterone were 115 +/- 39, 8,142 +/- 1,362, 10,070 +/- 1,999, and 7.7 +/- 1.5 microg/h, respectively. Urinary excretion rates of all metabolites were greater in both the 100 and 300 mg groups than in controls (P < 0.0001). Urinary excretion rates of testosterone (P = 0.007), androsterone (P = 0.009), etiocholanolone (P = 0.0005), and dihydrotestosterone (P < 0.0001) were greater in the subjects who received 300 mg androstenedione than in those who received 100 mg. In the treated groups, excretion of free testosterone accounted for less than 0.1% of the total excreted testosterone measured. Serum testosterone glucuronide levels increased significantly during frequent blood sampling in both the 100 and 300 mg groups compared with controls (P = 0.0005 for the 100 mg group; P < 0.0001 for the 300 mg group). The net mean changes in area under the curve for serum testosterone glucuronide were -18 +/- 25%, 579 +/- 572%, and 1267 +/- 1675% in the groups receiving 0, 100, and 300 mg/d androstenedione, respectively. We conclude that the administration of both 100 and 300 mg androstenedione increases the excretion rates of conjugated testosterone, androsterone, etiocholanolone, and dihydrotestosterone and the serum levels of testosterone glucuronide in men. The magnitude of these increases is much greater than the changes observed in serum total testosterone concentrations. These findings demonstrate that orally administered androstenedione is largely metabolized to testosterone glucuronide and other androgen metabolites before release into the general circulation. PMID- 11502793 TI - Improved glycemic control increases fasting plasma acylation-stimulating protein and decreases leptin concentrations in type II diabetic subjects. AB - Acylation-stimulating protein is an adipocyte-derived protein that has recently been suggested to play an important role in the regulation of triglyceride storage. To date, little information has been reported with regard to fasting acylation-stimulating protein levels and its relation to metabolic control, leptin, and/or lipids in subjects with diabetes mellitus. We therefore evaluated fasting acylation-stimulating protein, leptin, and lipid levels before and 4 months after improving glycemic control with sulfonylurea treatment in a group of poorly controlled obese women with type 2 diabetes and in age- and body mass index-matched nondiabetic obese women. Fasting plasma acylation-stimulating protein (49.67 +/- 19.73 vs. 48.49 +/- 20.70 nmol/liter) and leptin concentrations (33.7 +/- 23.2 vs. 26.2 +/- 10.6 ng/ml) were not significantly different between the groups. Improvement of glycemic control produced parallel falls in fasting blood glucose and hemoglobin A1c. Plasma leptin concentrations were also significantly reduced (33.69 +/- 23.2 vs. 22.73 +/- 11.26 ng/ml; P = 0.036), whereas fasting acylation-stimulating protein concentrations were significantly increased after treatment (48.49 +/- 20.70 vs. 72,82 +/- 29,72 nmol/liter; P = 0.004). Nevertheless, lipids and apolipoprotein B did not significantly improve. We could not find any correlation between elevated acylation-stimulating protein levels and changes in body mass index, glucose, insulin, hemoglobin A1c, leptin, or lipid levels. Similarly, the decrement in circulating leptin levels observed after treatment did not correlate with changes in the levels of glucose, insulin, hemoglobin A1c, or any lipid parameters. We conclude that improved glycemic control increases fasting acylation-stimulating protein and decreases leptin concentrations, but not corrects critical lipid abnormalities in type 2 obese diabetic subjects. Moreover, altered plasma acylation-stimulating protein levels are not associated with changes in body mass index or lipid, leptin, insulin, or glucose levels. Thus, our findings suggest that improved glycemic control or insulin resistance is not responsible for abnormal fatty acid trapping, and failure of lipids to improve after treatment in our patients is consistent with the acylation-stimulating protein resistance concept. PMID- 11502794 TI - Stimulation of human extravillous trophoblast migration by IGF-II is mediated by IGF type 2 receptor involving inhibitory G protein(s) and phosphorylation of MAPK. AB - We have earlier shown that migration and invasiveness of first trimester human extravillous trophoblast cells are stimulated by IGF-II, independently of IGF type 1 receptor and that migration stimulation is the primary reason for increased extravillous trophoblast cell invasiveness induced by IGF-II. In the present study we examined the functional role of IGF type II receptor in IGF-II stimulation of extravillous trophoblast cell migration and the underlying signal transduction pathways including the participation of inhibitory G protein(s) and MAPK. The migratory ability of a well characterized in vitro propagated human first trimester extravillous trophoblast cell line expressing the phenotype of extravillous trophoblast cells in situ was quantitated with a Transwell migration assay under different experimental conditions. We found that the extravillous trophoblast cells expressed an abundance of IGF type 2 receptor as detected by immunostaining and Western blots, and recombinant human IGF-II promoted their migration in a dose- and time-dependent manner. Both polyclonal and monoclonal IGF type 2 receptor-blocking antibodies blocked migration-stimulating effects of IGF-II. Two synthetic IGF-II analogs ([Leu27]IGF-II, which can bind to IGF type 2 receptor and IGF-binding proteins, but not IGF type 1 receptor, and [QAYL Leu27]IGF-II, which can bind to IGFR-II, but neither IGFR-I nor IGF-binding proteins) both stimulated extravillous trophoblast cell migration to levels higher than those induced by wild-type IGF-II. These results reveal that IGF-II action was mediated by IGF type 2 receptor, independently of IGF type 1 receptor and IGF-binding proteins. Treatment of extravillous trophoblast cell membrane preparations with IGF-II decreased adenylyl cyclase activity in a concentration dependant manner, indicating the participation of inhibitory G proteins in IGF-II action. This was substantiated further with the findings that increasing intracellular cAMP using forskolin or (Bu)2cAMP inhibited basal extravillous trophoblast cell migration and blocked IGF-II stimulation of migration. IGF-II treatment rapidly stimulated phosphorylation of MAPK (ERK-1 and -2), which was blocked by pretreatment of extravillous trophoblast cells with the MAPK kinase (MEK) inhibitor PD98059. Treatment with this inhibitor also blocked extravillous trophoblast cell migration in the presence or absence of IGF-II. These results, taken together, reveal that IGF-II stimulates extravillous trophoblast cell migration by signaling through IGF type 2 receptor, involving inhibitory G proteins and activating the MAPK pathway. PMID- 11502795 TI - Characterization of human FSH isoforms reveals a nonglycosylated beta-subunit in addition to the conventional glycosylated beta-subunit. AB - Human FSH consists of a mixture of isoforms that can be separated on the basis of differences in negative charge conferred by variations in the numbers of sialic acid residues that terminate oligosaccharide branches. Western analysis of human FSH isoforms separated by chromatofocusing revealed the presence of two human FSHbeta isoforms that differed in size. A low mol wt human FSHbeta isoform was associated with all FSH isoform fractions. A high mol wt human FSHbeta isoform was associated with the more acidic fractions and increased in relative abundance as the pI decreased. Characterization of representative human FSHbeta isoforms by mass spectrometry and automated Edman degradation revealed a low mol wt isoform that was not glycosylated. A high mol wt isoform was N-glycosylated at Asn residues 7 and 24. These results indicate that pituitary human FSH consists of two classes of molecules: those that possess a nonglycosylated beta-subunit and those that possess a glycosylated beta-subunit. Glycoprotein hormones are known to be elliptical molecules, and the beta-subunit oligosaccharides project outward from the short diameter, thereby increasing it. It is interesting to speculate that this change in shape might affect ultrafiltration rates, leading to differences in delivery rates to target tissues and elimination by filtration in the kidney. PMID- 11502796 TI - The role of IGF-binding proteins in mediating the effects of recombinant human IGF-I on insulin requirements in type 1 diabetes mellitus. AB - To determine the role of IGF-binding proteins in mediating the direct effects of recombinant human IGF-I on insulin requirements in type 1(insulin-dependent) diabetes mellitus, overnight changes in IGF-I, IGF-II, and IGF-binding protein-1, -2, and -3, collected under euglycemic conditions, were compared in nine subjects after double blind, randomized, sc administration of recombinant human IGF-I (40 microg/kg) or placebo at 1800 h. On both nights a somatostatin analog infusion (300 ng/kg x h) suppressed endogenous GH production, and three timed discrete GH pulses (total, 0.029 IU/kg x night) ensured identical GH levels. After recombinant human IGF-I administration, IGF-I levels and the IGF-I/IGF-binding protein-3 ratio increased [mean +/- SEM:IGF-I, 401 +/- 22 ng/ml; placebo, 256 +/- 20 ng/ml (P = 0.0002); IGF-I, 0.108 +/- 0.006; placebo, 0.074 +/- 0.004 (P = 0.0003), respectively], and insulin requirements decreased (IGF-I, 0.12 +/- 0.03; placebo, 0.23 +/- 0.03 U/kg x min; P = 0.008). The normal within-individual inverse relationships between insulin and IGF-binding protein-1 levels were observed (lag time 2 h: r = -0.34; P < 0.01). Yet despite reduced free insulin levels (8.5 +/- 1.5; placebo, 12.2 +/- 1.2 mU/liter; P = 0.03), IGF-binding protein-1 levels were reduced after recombinant human IGF-I administration (53.7 +/- 6.8; placebo, 82.2 +/- 11.8 ng/ml; P = 0.008). The largest reductions in free insulin levels after recombinant human IGF-I and thus putative improvement in insulin sensitivity occurred in subjects with the smallest increase in the plasma IGF-I/IGF-binding protein-3 ratio (r = 0.7; P = 0.03). Taken together, these data are consistent with the hypothesis that transcapillary movement of IGF-I (perhaps mediated by IGF-binding protein-1), out of the circulation facilitates altered insulin sensitivity. These data have important implications for risk-benefit assessment of recombinant human IGF-I therapy in type 1 diabetes mellitus. PMID- 11502797 TI - Familial corticosteroid-binding globulin deficiency due to a novel null mutation: association with fatigue and relative hypotension. AB - Corticosteroid-binding globulin is a 383-amino acid glycoprotein that serves a hormone transport role and may have functions related to the stress response and inflammation. We describe a 39-member Italian-Australian family with a novel complete loss of function (null) mutation of the corticosteroid-binding globulin gene. A second, previously described, mutation (Lyon) segregated independently in the same kindred. The novel exon 2 mutation led to a premature termination codon corresponding to residue -12 of the procorticosteroid-binding globulin molecule (c.121G-->A). Among 32 family members there were 3 null homozygotes, 19 null heterozygotes, 2 compound heterozygotes, 3 Lyon heterozygotes, and 5 individuals without corticosteroid-binding globulin mutations. Plasma immunoreactive corticosteroid-binding globulin was undetectable in null homozygotes, and mean corticosteroid-binding globulin levels were reduced by approximately 50% at 18.7 +/- 1.3 microg/ml (reference range, 30-52 microg/ml) in null heterozygotes. Morning total plasma cortisol levels were less than 1.8 microg/dl in homozygotes and were positively correlated to the plasma corticosteroid-binding globulin level in heterozygotes. Homozygotes and heterozygote null mutation subjects had a high prevalence of hypotension and fatigue. Among 19 adults with the null mutation, the systolic blood pressure z-score was 12.1 +/- 3.5; 11 of 19 subjects (54%) had a systolic blood pressure below the third percentile. The mean diastolic blood pressure z-score was 18.1 +/- 3.4; 8 of 19 subjects (42%) had a diastolic blood pressure z-score below 10. Idiopathic chronic fatigue was present in 12 of 14 adult null heterozygote subjects (86%) and in 2 of 3 null homozygotes. Five cases met the Centers for Disease Control criteria for chronic fatigue syndrome. Fatigue questionnaires revealed scores of 25.1 +/- 2.5 in 18 adults with the mutation vs. 4.2 +/- 1.5 in 23 healthy controls (P < 0.0001). Compound heterozygosity for both mutations resulted in plasma cortisol levels comparable to those in null homozygotes. Abnormal corticosteroid-binding globulin concentrations or binding affinity may lead to the misdiagnosis of isolated ACTH deficiency. The mechanism of the association between fatigue and relative hypotension is not established by these studies. As idiopathic fatigue disorders are associated with relatively low plasma cortisol, abnormalities of corticosteroid-binding globulin may be pathogenic. PMID- 11502798 TI - A comprehensive analysis of MNG1, TCO1, fPTC, PTEN, TSHR, and TRKA in familial nonmedullary thyroid cancer: confirmation of linkage to TCO1. AB - About 5% of nonmedullary thyroid cancer is familial. These familial nonmedullary thyroid cancer cases are characterized by an earlier age of onset, more aggressive phenotype, and in some families a high propensity to benign thyroid disease. Little is known about the genes conferring predisposition to nonmedullary thyroid cancer. Three loci have been identified through genetic linkage: MNG1 on 14q32, TCO1 on 19p13.2, and fPTC on 1p21. In addition to these putative genes, a number of loci represent candidate familial nonmedullary thyroid cancer predisposition genes by virtue of their involvement in sporadic disease (TRKA), their role in benign disease (TSHR), and because they underlie syndromes with a risk of nonmedullary thyroid cancer (PTEN). To evaluate the roles of MNG1, TCO1, fPTC, PTEN, TSHR, and TRKA in familial nonmedullary thyroid cancer, we have carried out a comprehensive mutation and linkage analysis of these genes in 22 families. One family was linked to chromosome 19q13.2, confirming that TCO1 underlies a subset of familial nonmedullary thyroid cancer. None of the families was linked to MNG1 or fPTC, and there was no evidence to support the roles of PTEN, TSHR, or TRKA. Familial nonmedullary thyroid cancer is an emerging clinical phenotype that is genetically heterogeneous, and none of the currently identified genes accounts for the majority of families. PMID- 11502799 TI - Class III alleles of the variable number of tandem repeat insulin polymorphism associated with silencing of thymic insulin predispose to type 1 diabetes. AB - Type 1 diabetes results from autoimmune destruction of the insulin-producing pancreatic beta cells. The insulin gene (INS) is also expressed in human thymus, an ectopic expression site likely involved in immune tolerance. The IDDM2 diabetes susceptibility locus maps to a minisatellite composed of a variable number of tandem repeats situated 0.5 kb upstream of INS. Chromosomes carrying the protective long INS variable number of tandem repeats alleles (class III) produce higher levels of thymic INS mRNA than those with the predisposing, short class I alleles. However, complete silencing of thymic INS transcripts from the class III chromosome was found in a small proportion of heterozygous human thymus samples. We hypothesized that the specific class III alleles found on these chromosomes silence rather than enhance thymic insulin expression. To test the prediction that these alleles are predisposing, we developed a DNA fingerprinting method for detecting two putative "silencing" alleles found in two thymus samples (S1, S2). In a set of 287 diabetic children and their parents we found 13 alleles matching the fingerprint of the S1 or S2 alleles. Of 18 possible transmissions, 12 of the S1-S2 alleles were transmitted to the diabetic offspring, a frequency of 0.67, significantly higher than the 0.38 seen in the remaining 142 class III alleles; P = 0.025. This confirms our prediction and represents an additional level of correlation between thymic insulin and diabetes susceptibility, which supports a thymic enhancer effect of the INS variable number of tandem repeats as the mechanism of IDDM2 and refines the contribution of IDDM2 genotyping to diabetes risk assessment. PMID- 11502800 TI - Behavior of small thyroid cancers found by screening radiation-exposed individuals. AB - Thyroid cancers detected by screening irradiated individuals are often small and of uncertain clinical significance. We retrospectively analyzed the effect of screening in a cohort of 4296 individuals exposed to radiation as children in the 1940s and 1950s and followed by us from 1974 until the present. We compared the thyroid cancers diagnosed before 1974 (122 cases, routine care) with the cancers found in subjects screened by us after 1974 (172 cases, screened), using cancer recurrence as the end point. Screening included a thyroid scan or, more recently, thyroid ultrasound. As expected, many of the cancers found by screening were very small (52% were <10 mm), but the range of tumor sizes overlapped those found by routine care. The recurrence rate was significantly lower in the cases found by screening, but when the comparison was limited to cancers 10 mm or larger, no difference in the recurrence rates was seen. This would suggest that the lower recurrence rate observed for small thyroid cancers detected at screening was due to earlier diagnosis rather than more effective treatment. By univariate analysis, four factors were associated with an increased risk of recurrence of small (<10 mm) thyroid cancers: short latency (i.e. a shorter time interval between the radiation exposure and the first thyroid surgery), lymph node metastases present at diagnosis, multifocal cancers, and higher radiation dose. In a multivariate analysis combining the four risk factors, only short latency was significant. As thyroid cancers that escape detection by routine means should be diagnosed at screening, and both large and small thyroid cancers have the potential to recur, screening may be of value, but only if groups with a sufficiently high prevalence of thyroid cancer can be identified to offset the adverse effects of unnecessary treatment due to false positive results. PMID- 11502801 TI - Determinants of the impaired secretion of glucagon-like peptide-1 in type 2 diabetic patients. AB - To elucidate the causes of the diminished incretin effect in type 2 diabetes mellitus we investigated the secretion of the incretin hormones, glucagon-like peptide-1 and glucose- dependent insulinotropic polypeptide and measured nonesterified fatty acids, and plasma concentrations of insulin, C peptide, pancreatic polypeptide, and glucose during a 4-h mixed meal test in 54 heterogeneous type 2 diabetic patients, 33 matched control subjects with normal glucose tolerance, and 15 unmatched subjects with impaired glucose tolerance. The glucagon-like peptide-1 response in terms of area under the curve from 0-240 min after the start of the meal was significantly decreased in the patients (2482 +/- 145 compared with 3101 +/- 198 pmol/liter.240 min; P = 0.024). In addition, the area under the curve for glucose-dependent insulinotropic polypeptide was slightly decreased. In a multiple regression analysis, a model with diabetes, body mass index, male sex, insulin area under the curve (negative influence), glucose-dependent insulinotropic polypeptide area under the curve (negative influence), and glucagon area under the curve (positive influence) explained 42% of the variability of the glucagon-like peptide-1 response. The impaired glucose tolerance subjects were hyperinsulinemic and generally showed the same abnormalities as the diabetic patients, but to a lesser degree. We conclude that the meal-related glucagon-like peptide-1 response in type 2 diabetes is decreased, which may contribute to the decreased incretin effect in type 2 diabetes. PMID- 11502802 TI - Acute insulin responses to leucine in children with the hyperinsulinism/hyperammonemia syndrome. AB - Mutations of glutamate dehydrogenase cause the hyperinsulinism/hyperammonemia syndrome by desensitizing glutamate dehydrogenase to allosteric inhibition by GTP. Normal allosteric activation of glutamate dehydrogenase by leucine is thus uninhibited, leading us to propose that children with hyperinsulinism/hyperammonemia syndrome will have exaggerated acute insulin responses to leucine in the postabsorptive state. As hyperglycemia increases beta cell GTP, we also postulated that high glucose concentrations would extinguish abnormal responsiveness to leucine in hyperinsulinism/hyperammonemia syndrome patients. After an overnight fast, seven hyperinsulinism/hyperammonemia syndrome patients (aged 9 months to 29 yr) had acute insulin responses to leucine performed using an iv bolus of L-leucine (15 mg/kg) administered over 1 min and plasma insulin measurements obtained at -10, -5, 0, 1, 3, and 5 min. The acute insulin response to leucine was defined as the mean increase in insulin from baseline at 1 and 3 min after an iv leucine bolus. The hyperinsulinism/hyperammonemia syndrome group had excessively increased insulin responses to leucine (mean +/- SEM, 73 +/- 21 microIU/ml) compared with the control children and adults (n = 17) who had no response to leucine (1.9 +/- 2.7 microU/ml; P < 0.05). Four hyperinsulinism/hyperammonemia syndrome patients then had acute insulin responses to leucine repeated at hyperglycemia (blood glucose, 150-180 mg/dl). High blood glucose suppressed their abnormal baseline acute insulin responses to leucine of 180, 98, 47, and 28 microU/ml to 73, 0, 6, and 19 microU/ml, respectively. This suppression suggests that protein-induced hypoglycemia in hyperinsulinism/hyperammonemia syndrome patients may be prevented by carbohydrate loading before protein consumption. PMID- 11502803 TI - Leptin, cortisol, and GH secretion interactions in short normal prepubertal children. AB - The hormonal regulation of the ob gene and leptin secretion in humans is still unclear. To investigate the interactions among leptin, cortisol, and GH, we analyzed and time-cross-correlated their spontaneous 24-h secretion in 12 short normal prepubertal children of both sexes (6 females and 6 males). Time-cross correlation analyses demonstrated that leptin and cortisol were correlated in both a negative and positive fashion. The negative correlation, with cortisol leading leptin by 4 and 3 h for boys and girls, respectively, might reflect the stimulatory effect of CRH on the sympathetic system, which inhibits leptin secretion; the positive correlation, with leptin leading cortisol by 6 and 5 h for boys and girls, respectively, might reflect a direct effect of leptin on CRH secretion in the hypophyseal portal system. Time-cross-correlation analyses also showed a strong positive correlation between GH and leptin concentrations, with GH leading leptin by 5 and 2 h for boys and girls, respectively, suggesting a possible direct leptin-releasing effect of GH on adipocytes. We conclude that cross-correlation analyses of 24-h hormone secretions under baseline physiological conditions suggest that the hypothalamic-pituitary-adrenal axis might have a prevailing inhibitory effect on leptin secretion, whereas leptin might exert a positive effect on the hypothalamic-pituitary-adrenal axis. The relation between GH and leptin could be a direct one and characterized prevalently by a positive effect of GH on leptin secretion. Further investigations using different experimental systems are needed to ascertain the validity of these mathematically educed conclusions. PMID- 11502804 TI - Is population bone mineral density variation linked to the marker D11S987 on chromosome 11q12-13? AB - Our purpose is to test linkage of human chromosome 11q12-13 to BMD variation. Chromosome 11q12-13 has been linked to three BMD-related phenotypes that are inherited as Mendelian traits in human pedigrees: an autosomal dominant high bone mass trait, autosomal recessive osteoporosis pseudoglioma, and autosomal recessive osteopetrosis. A sibling pair study with 374 sibships showed significant linkage of D11S987 to normal BMD variation, with a maximum logarithm of odds score of 3.5. However, a subsequent linkage study with a total of 595 sibling pairs demonstrated reduced significance for linkage of D11S987 to bone mineral density variation, with a logarithm of odds score less than 2.2. We genotyped five markers in a genomic region of approximately 27 cM centering on D11S987 and measured bone mineral density and other traits (weight, etc.) for 635 individuals from 53 human pedigrees. Each of these pedigrees was ascertained through a proband with bone mineral density Z-scores less than -1.28 at the hip or spine. Adjusting for age, sex, and weight as covariates, we performed two point and multipoint linkage analyses using the variance component linkage analysis method implemented in Sequential Oligogenic Linkage Analysis Routines. We found little evidence of linkage of these five markers to bone mineral density at the spine, hip, wrist and total body bone mineral content. The maximum logarithm of odds score at these five markers was 0.25, and the maximum logarithm of odds score at D11S987 was 0.15. Therefore, although we cannot exclude the linkage of D11S987 region to bone mineral density variation, there is no evidence for linkage of the marker D11S987 on human chromosome 11q12-13 to bone mineral density variation in our study population. PMID- 11502805 TI - Effect of glucocorticoid therapy on energy intake in children treated for acute lymphoblastic leukemia. AB - Despite a widespread belief that glucocorticoid therapy is associated with positive energy balance and excess weight gain there is a dearth of quantitative evidence about its effects and the underlying mechanisms of any effects. The primary aim of the present study was to quantify the effect of dexamethasone and prednisone treatment on energy intake in children treated for childhood acute lymphoblastic leukemia. A secondary aim was to test for differences in excess weight gain between patients treated using the 2 glucocorticoids. We measured energy intake in 26 patients (mean +/- SD age, 6.3 +/- 2.3 yr) during a 5-d period "on" steroids and again in the week before steroid treatment. Changes in body mass index from diagnosis to 1 and 2 yr postdiagnosis were expressed as SD scores. Steroid treatment was associated with a significant increase in energy intake of approximately 20% (mean paired difference, 1.7 MJ/d; SD, 2.8; 95% confidence interval, 0.7-2.8 MJ/d), with no significant difference between the 2 steroids. The mean change in body mass index SD score was +0.38 (SD, 1.10; P < 0.05) to 1 yr and +0.68 (SD, 1.38; P < 0.05) to 2 yr, with no significant difference between the 2 groups of patients. Glucocorticoid treatment in childhood acute lymphoblastic leukemia increases energy intake markedly, and this effect contributes to the excess weight gain and obesity characteristic of patients being treated for acute lymphoblastic leukemia. PMID- 11502806 TI - Familial medullary thyroid carcinoma with noncysteine ret mutations: phenotype genotype relationship in a large series of patients. AB - Familial medullary thyroid carcinoma only is related to germline mutations in the protooncogene RET, mainly in exons 10, whereas noncysteine mutations (exons 13 15) are considered infrequent. We analyzed 148 patients from 47 familial medullary thyroid carcinoma only families, and we found noncysteine RET mutations in 59.5% of these families. Of the index cases with noncysteine mutations, 43.4% presented with a multinodular goiter and high basal calcitonin; they were older at diagnosis than those with mutation in exon 10 and had more multifocal medullary thyroid carcinoma, but no difference in size, bilaterality, presence of C cell hyperplasia, or nodal metastases was found. Gene carriers with noncysteine RET mutations had a lower incidence of medullary thyroid carcinoma (78.2% vs. 94.1%) than those with mutation in exon 10; 20.2% had C cell hyperplasia only, although thyroidectomized at an older age. In conclusion, familial medullary thyroid carcinoma with noncysteine RET mutations are not infrequent and are overrepresented in presumed sporadic medullary thyroid carcinoma, suggesting that RET analysis should routinely be extended to exons 13, 14, and 15. The phenotype is characterized by a late onset of the disease, suggesting a delayed appearance of C cell disease rather than a less aggressive form. In familial medullary thyroid carcinoma gene carriers, the optimal timing for thyroidectomy remains controversial. Based on these data, we propose that surgery should be performed before elevation of the basal calcitonin level, potentially as soon as the pentagastrin test becomes abnormal. PMID- 11502807 TI - Two distinct MICA gene markers discriminate major autoimmune diabetes types. AB - The polymorphism of the major histocompatibility complex class I chain-related A gene is associated with type 1 diabetes mellitus. The major histocompatibility complex class I chain-related A gene 5 allele is significantly more frequent in Caucasian type 1 diabetes mellitus children than in healthy subjects, but no information is available on the association with adult-onset type 1 diabetes mellitus or with the so-called slowly progressive latent autoimmune diabetes of the adult in the same ethnic group. In this study we estimated the frequency of major histocompatibility complex class I chain- related A gene alleles and human leukocyte antigen-DRB1*03-DQA1*0501-DQB1*0201 and human leukocyte antigen-DRB1*04 DQA1*0301-DQB1*0302 in 195 type 1 diabetes mellitus subjects, in 80 latent autoimmune diabetes of the adult subjects, and in 158 healthy subjects from central Italy. Major histocompatibility complex class I chain-related A gene 5 was significantly associated with type 1 diabetes mellitus only in the 1-25 yr age group at diagnosis, and the odds ratio of the simultaneous presence of both major histocompatibility complex class I chain-related A gene 5 and human leukocyte antigen-DRB1*03- DQA1*0501-DQB1*0201 and/or human leukocyte antigen DRB1*04-DQA1*0301-DQB1*0302 was as high as 54 and higher than 388 when compared with double negative individuals. Adult-onset type 1 diabetes mellitus (age at diagnosis, >25 yr) and latent autoimmune diabetes of the adult were significantly associated with major histocompatibility complex class I chain-related A gene 5.1, which was not significantly increased among diabetic children. Only the combination of major histocompatibility complex class I chain-related A gene 5.1 and human leukocyte antigen-DRB1*03-DQA1*0501-DQB1*0201 and/or human leukocyte antigen-DRB1*04-DQA1*0301-DQB1*0302 conferred increased risk for adult-onset type 1 diabetes mellitus or for latent autoimmune diabetes of the adult. Our study provides demonstration of the existence of distinct genetic markers for childhood/young-onset type 1 diabetes mellitus and for adult-onset type 1 diabetes mellitus/latent autoimmune diabetes of the adult, namely major histocompatibility complex class I chain-related A gene 5 and major histocompatibility complex class I chain-related A gene 5.1, respectively. PMID- 11502808 TI - TNF-alpha and hyperandrogenism: a clinical, biochemical, and molecular genetic study. AB - To evaluate the role of TNF-alpha in the pathogenesis of hyperandrogenism, we have evaluated the serum TNF-alpha levels, as well as several polymorphisms in the promoter region of the TNF-alpha gene, in a group of 60 hyperandrogenic patients and 27 healthy controls matched for body mass index. Hyperandrogenic patients presented with mildly increased serum TNF-alpha levels as compared with controls (mean[median] +/- SD: 7.2[7.0] +/- 3.3 pg/ml vs. 5.6[4.4] +/- 4.0 pg/ml, P < 0.02). Although no differences in body mass index and insulin resistance indexes were observed between patients and controls, when subjects were classified by body weight, serum TNF-alpha was increased only in lean patients as compared with lean controls, but this difference was not statistically significant when comparing obese patients with obese controls. The TNF-alpha gene polymorphisms studied here (-1196C/T, -1125G/C, -1031T/C, -863C/A, -857C/T, 316G/A, -308G/A, -238G/A, and -163G/A) were equally distributed in hyperandrogenic patients and controls. However, carriers of the -308A variant presented with increased basal and leuprolide-stimulated serum androgens and 17 hydroxyprogesterone levels when considering patients and controls as a group. No differences were observed in serum TNF-alpha levels, body mass index, and insulin resistance indexes, depending on the presence or absence of these variants. In conclusion, our present results suggest that the TNF-alpha system might contribute to the pathogenesis of hyperandrogenism, independent of obesity and insulin resistance. However, elucidation of the precise mechanisms underlying the relationship between the TNF-alpha system and androgen excess is needed before considering TNF-alpha as a significant contributing factor to the development of hyperandrogenism. PMID- 11502809 TI - Effect of short-term fasting on free and bound leptin concentrations in lean and obese women. AB - Plasma leptin exists in protein-bound and free forms, which may affect its hormonal bioactivity. Therefore, the relationship between bound and free leptin may be particularly important during physiological conditions that cause rapid alterations in total plasma leptin concentration, such as fasting. The purpose of this study was to evaluate the effect of short-term fasting on bound and free plasma leptin concentrations and leptin binding capacity (a measure of plasma leptin-binding protein content) in lean and obese women. Six lean (body mass index, 21 +/- 1 kg/m2) and 6 abdominally obese (BMI, 36 +/- 1 kg/m2) women were studied after 14 h and 22 h of fasting. Although total plasma leptin concentration was more than 6-fold greater in obese (45.4 +/- 7.6 microg/liter) compared with lean (7.4 +/- 1.0 microg/liter) women at 14 h of fasting (P < 0.05), the percentage of leptin in the bound form was greater in lean than obese subjects (29 +/- 2% vs. 12 +/- 3%; P < 0.05). Arterial total, free, and bound plasma leptin concentrations all declined between 14 h and 22 h of fasting in both lean and obese groups, but the relative decline of these fractions was greater in lean (36 +/- 4%, 60 +/- 9%, and 51 +/- 13%, respectively) than in obese (19 +/- 5%, 21 +/- 8%, and 12 +/- 7%, respectively) subjects (all P < 0.05). In contrast, leptin binding capacity was unchanged. The percentage of total plasma leptin present in bound form was constant between 14 h and 22 h of fasting in lean subjects and increased slightly but significantly in obese subjects. These data demonstrate that both free and bound fractions of leptin in plasma decrease quickly in response to energy restriction, but the decline is blunted in abdominally obese compared with lean women. In addition, the equilibrium between bound and free leptin fractions is maintained during brief fasting and is not regulated by leptin binding capacity. PMID- 11502810 TI - Disruption of the synchronous secretion of leptin, LH, and ovarian androgens in nonobese adolescents with the polycystic ovarian syndrome. AB - The present study probes putative disruption of hypothalamic control of multihormone outflow in polycystic ovarian syndrome by quantitating the joint synchrony of leptin and LH release in adolescents with this syndrome and eumenorrheic controls. To this end, hyperandrogenemic oligo- or anovulatory patients with polycystic ovarian syndrome (n = 11) and healthy girls (n = 9) underwent overnight blood sampling every 20 min for 12 h to monitor simultaneous secretion of leptin (immuno-radiometric assay), LH (immunofluorometry), and androstenedione and T (RIA). Synchronicity of paired leptin-LH, leptin androstenedione, and leptin-T profiles was appraised by two independent bivariate statistics; viz., lag-specific cross-correlation analysis and pattern-sensitive cross-approximate entropy. The study groups were comparable in chronological and postmenarchal age, body mass index, fasting plasma insulin/glucose ratios, and serum E2 concentrations. Overnight mean (+/- SEM) serum leptin concentrations were not distinguishable in the two study groups at 30 +/- 4.8 (polycystic ovarian syndrome) and 32 +/- 7.4 microg/liter (control). Serum LH concentrations were elevated at 9.5 +/- 1.4 in girls with polycystic ovarian syndrome vs. 2.8 +/ 0.36 IU/liter in healthy subjects (P = 0.0015), androstenedione at 2.8 +/- 0.30 (polycystic ovarian syndrome) vs. 1.2 +/- 0.11 ng/ml (control) (P = 0.0002), and T at 1.56 +/- 0.29 (polycystic ovarian syndrome) vs. 0.42 +/- 0.06 ng/ml (P < 0.0001). Cross-correlation analysis shows that healthy adolescents maintained a positive relationship between leptin and LH release, wherein the latter lagged by 20 min (P < 0.01). No such association emerged in girls with polycystic ovarian syndrome. In eumenorrheic volunteers, leptin and androstenedione concentrations also covaried in a lag-specific manner (0.0001 < P < 0.01), but this linkage was disrupted in patients with polycystic ovarian syndrome. Anovulatory adolescents further failed to sustain normal time-lagged coupling between leptin and T (P < 0.01). Approximate entropy calculations revealed erosion of orderly patterns of leptin release in polycystic ovarian syndrome (P = 0.012 vs. control). Cross entropy analysis of two-hormone pattern regularity disclosed marked disruption of leptin and LH (P = 0.0099), androstenedione and leptin (P = 0.0075) and T-leptin (P = 0.019) synchrony in girls with polycystic ovarian syndrome. In summary, hyperandrogenemic nonobese adolescents with oligo- or anovulatory polycystic ovarian syndrome manifest: 1) abrogation of the regularity of monohormonal leptin secretory patterns, despite normal mean serum leptin concentrations; 2) loss of the bihormonal synchrony between leptin and LH release; and 3) attenuation of coordinate leptin and androstenedione as well as leptin and T output. In ensemble, polycystic ovarian syndrome pathophysiology in lean adolescents is marked by vivid impairment of the synchronous outflow of leptin, LH and androgens. Whether analogous disruption of leptin-gonadal axis integration is ameliorated by therapy and/or persists into adulthood is not known. PMID- 11502811 TI - Diagnostic accuracy of 18F-fluorodeoxyglucose positron emission tomography in the follow-up of papillary or follicular thyroid cancer. AB - Positron emission tomography with 18F-fluorodeoxyglucose is a relatively new nuclear imaging technique in oncology. We conducted a systematic review to determine the diagnostic accuracy of 18F-fluorodeoxyglucose positron emission tomography in patients suspected of recurrent papillary or follicular thyroid carcinoma. Two reviewers independently selected, extracted, and assessed data from relevant literature found in computerized databases and by reference tracking. Prospective and retrospective studies with 10 human subjects, or more, that evaluated the accuracy of ring positron emission tomography, using 18F fluorodeoxyglucose in follicular and papillary thyroid cancer, were included. Studies on 18F-fluorodeoxyglucose imaging using gamma cameras, reviews, case reports, editorials, letters, and comments were excluded. The methodological quality was assessed by applying the criteria for diagnostic tests recommended by the Cochrane Methods Group on Screening and Diagnostic Tests. A rating system was used for qualitative analysis consisting of four levels of evidence (1 = highest level; 4 = lowest level). Fourteen studies met the inclusion criteria. All studies claimed a positive role for positron emission tomography but, at evidence levels 3 or 4, precluding quantitative analysis. Methodological problems included poor validity of reference tests and a lack of blinding of test performance and interpretation. The reviewed material was heterogeneous with respect to patient variation and validation methodology. The most consistent data were found on the ability of 18F-fluorodeoxyglucose positron emission tomography to provide an anatomical substrate in patients with elevated serum Tg and negative iodine-131 scans. In conclusion, the results seem to support the potential of 18F fluorodeoxyglucose positron emission tomography to identify and localize foci of recurrent cancer in the latter patient subset. However, implementation of positron emission tomography in a routine diagnostic algorithm requires additional evidence. PMID- 11502812 TI - Chronic insomnia is associated with nyctohemeral activation of the hypothalamic pituitary-adrenal axis: clinical implications. AB - Although insomnia is, by far, the most commonly encountered sleep disorder in medical practice, our knowledge in regard to its neurobiology and medical significance is limited. Activation of the hypothalamic-pituitary-adrenal axis leads to arousal and sleeplessness in animals and humans; however, there is a paucity of data regarding the activity of the hypothalamic-pituitary-adrenal axis in insomniacs. We hypothesized that chronic insomnia is associated with increased plasma levels of ACTH and cortisol. Eleven young insomniacs (6 men and 5 women) and 13 healthy controls (9 men and 4 women) without sleep disturbances, matched for age and body mass index, were monitored in the sleep laboratory for 4 consecutive nights, whereas serial 24-h plasma measures of ACTH and cortisol were obtained during the fourth day. Insomniacs, compared with controls, slept poorly (significantly higher sleep latency and wake during baseline nights). The 24-h ACTH and cortisol secretions were significantly higher in insomniacs, compared with normal controls (4.2 +/- 0.3 vs. 3.3 +/- 0.3 pM, P = 0.04; and 218.0 +/- 11.0 vs. 190.4 +/- 8.3 nM, P = 0.07). Within the 24-h period, the greatest elevations were observed in the evening and first half of the night. Also, insomniacs with a high degree of objective sleep disturbance (% sleep time < 70), compared with those with a low degree of sleep disturbance, secreted a higher amount of cortisol. Pulsatile analysis revealed a significantly higher number of peaks per 24 h in insomniacs than in controls (P < 0.05), whereas cosinor analysis showed no differences in the temporal pattern of ACTH or cortisol secretion between insomniacs and controls. We conclude that insomnia is associated with an overall increase of ACTH and cortisol secretion, which, however, retains a normal circadian pattern. These findings are consistent with a disorder of central nervous system hyperarousal rather than one of sleep loss, which is usually associated with no change or decrease in cortisol secretion or a circadian disturbance. Chronic activation of the hypothalamic-pituitary-adrenal axis in insomnia suggests that insomniacs are at risk not only for mental disorders, i.e. chronic anxiety and depression, but also for significant medical morbidity associated with such activation. The therapeutic goal in insomnia should be to decrease the overall level of physiologic and emotional arousal, and not just to improve the nighttime sleep. PMID- 11502813 TI - Relevant cAMP-specific phosphodiesterase isoforms in human pituitary: effect of Gs(alpha) mutations. AB - Both cAMP production by adenylyl cyclase and cAMP degradation by phosphodiesterases account for intracellular cAMP levels. We previously demonstrated an increased phosphodiesterase activity in GH-secreting adenomas bearing the gsp oncogene. Here we characterize both the activity and the expression of cAMP-specific phosphodiesterase genes in the human pituitary and in gsp+ and gsp- GH-secreting adenomas and analyze the impact of this intracellular feedback mechanism on the levels of cAMP-responsive element-binding protein phosphorylation. Normal pituitary and gsp- GH-secreting adenomas showed similar phosphodiesterase activities, and 7-fold higher levels were observed in gsp+ tumors. In these tumors the increased activity was mainly owing to isobutyl methyl-xanthine-sensitive phosphodiesterase 4 and to isobutyl-methyl-xanthine insensitive isoforms. By semiquantitative RT-PCR, all phosphodiesterase 4 transcripts were expressed in the normal and tumoral pituitary. However, the levels of phosphodiesterase 4C and 4D messenger RNAs were significantly higher in gsp+ than in gsp- GH-secreting adenomas and normal pituitary. Expression of the thyroid-specific isobutyl-methyl-xanthine-insensitive phosphodiesterase 8B was absent in the normal pituitary but detectable in almost all GH-secreting adenomas and higher in gsp+ (P < 0.02). Therefore, this study provides a characterization of phosphodiesterase expression in human pituitary and demonstrates a dramatic induction of the cAMP-specific phosphodiesterases 4C and phosphodiesterases 4D and phosphodiesterases 8B in gsp+ GH-secreting adenomas. Similar levels of cAMP responsive element-binding protein phosphorylation were observed in gsp- and gsp+ GH-secreting adenomas; however, phosphodiesterase blockade caused an increase in cAMP-responsive element-binding protein phosphorylation that was significantly higher in gsp+ than in gsp- adenomas. Because cAMP-responsive element-binding protein represents the principal end point of the cAMP pathway, these results suggest that the enhanced phosphodiesterase activity may have a significant impact on the phenotypic expression of gsp mutations. PMID- 11502814 TI - Secondary hyperparathyroidism and bone turnover in elderly blacks and whites. AB - This study was undertaken to describe the prevalence of secondary hyperparathyroidism in African-American and Caucasian participants in the Boston Low-Income Elderly Osteoporosis Study and to examine and compare associations of hyperparathyroidism with biochemical markers of bone turnover and bone density in the two racial groups. Serum osteocalcin and serum cross-linked N-telopeptides of type I collagen, and calcaneal bone mineral density were measured in February or March in 255 men and women, 64 yr of age and older. Subjects were categorized as normal or as having hyperparathyroidism, based on a serum PTH concentration below or above the top of the normal range (6.9 pmol/liter), respectively. The prevalence of hyperparathyroidism was 38% in the 144 black subjects and 20% in the 111 white subjects. Serum osteocalcin and cross-linked N-telopeptides of type I collagen were significantly higher in both black and white hyperparathyroid subjects (P < 0.05), and the hyperparathyroid-related difference in osteocalcin was greater among black than white subjects. Hyperparathyroidism was significantly associated with reduced heel bone mineral density in blacks (P = 0.008) but not in whites. This study provides evidence that secondary hyperparathyroidism is prevalent in elderly adults, both black and white, and that it should not be viewed as a benign condition in either group. Recent public health efforts to promote higher calcium and vitamin D intakes, targeted predominantly to older Caucasians, should also be directed to older African Americans. PMID- 11502815 TI - Elevated plasma levels of soluble receptors of TNF-alpha and their association with smoking and microvascular complications in young adults with type 1 diabetes. AB - The purposes of this study were 1) to compare soluble tumor necrosis factor-alpha receptors, which are thought to reflect the degree of TNF-alpha activation, in nondiabetic subjects and type 1 diabetic patients, and 2) to evaluate the effects of smoking and microvascular complications on soluble tumor necrosis factor-alpha receptor levels in type 1 diabetic individuals. Plasma soluble tumor necrosis factor-alpha receptor levels (R1 and R2) were measured in 50 young type 1 diabetic patients without clinical macroangiopathy and in a matched group of 20 healthy volunteers. When diabetic patients were grouped according to smoking and microvascular complication status, the groups of patients had similar values of age, sex, body mass index, blood pressure, lipids, creatinine, and glycometabolic control. Nevertheless, soluble tumor necrosis factor-alpha receptor-R1 levels but not R2 levels, were markedly elevated (P < 0.05 or less) in complicated vs. uncomplicated (2.40 +/- 0.3 vs. 1.80 +/- 0.1 ng/ml) patients and in smokers vs. nonsmokers (2.66 +/- 0.4 vs. 1.76 +/- 0.1 ng/ml). In a two-factor ANOVA, both smoking (P < 0.01) and microvascular complications (P < 0.05) were independent predictors of soluble tumor necrosis factor-alpha receptor-R1. Soluble tumor necrosis factor-alpha receptor levels of diabetic patients who did not smoke or without complications were similar to those of healthy controls. In conclusion, smoking and microvascular complications seem to exert an additive and deleterious impact on TNF-alpha activation, as reflected by levels of soluble tumor necrosis factor-alpha receptors, in young adults with type 1 diabetes. PMID- 11502816 TI - Mutation of somatostatin receptor type 5 in an acromegalic patient resistant to somatostatin analog treatment. AB - Introduction of somatostatin analogs has greatly contributed to improving the prognosis of acromegaly. Although the majority of patients are effectively treated by these agents, resistance occurs in a subset of patients. So far, resistance to somatostatin has never been associated with mutations of the somatostatin receptor subtypes (sst2 and sst5) that inhibit GH secretion. Molecular analysis of genomic DNA from pituitary tumor and peripheral blood obtained from an acromegalic resistant to octreotide showed a somatic activating mutation of Gsalpha (Arg201Cys), no mutation in sst2, and one polymorphism (Pro109Ser) and one germ line mutation (Arg240Trp) in sst5. Wild-type (WT) and mutant sst5 PCR products were cloned and transfected into Chinese hamster ovary K1 cells. In Chinese hamster ovary K1 cells stably expressing mutant sst5, somatostatin-28 was less potent in inhibiting cyclic AMP levels than in WT cells. Proliferation of mutant cells exceeded that of WT by 50%. Moreover, somatostatin reduced cell growth and MAPK activity in WT but not in mutant cells in which the peptide even increased MAPK activity. We suggest that this mutation that abrogates the antiproliferative action of somatostatin and activates mitogenic pathways may be involved in the resistance to somatostatin treatment. PMID- 11502817 TI - Weight reduction increases plasma levels of an adipose-derived anti-inflammatory protein, adiponectin. AB - Adiponectin, an adipose tissue-specific plasma protein, was recently revealed to have anti-inflammatory effects on the cellular components of vascular wall. Its plasma levels were significantly lower in men than in women and lower in human subjects with obesity, type 2 diabetes mellitus, or coronary artery disease. Therefore, it may provide a biological link between obesity and obesity-related disorders such as atherosclerosis, against which it may confer protection. In this study, we observed the changes of plasma adiponectin levels with body weight reduction among 22 obese patients who received gastric partition surgery. A 46% increase of mean plasma adiponectin level was accompanied by a 21% reduction in mean body mass index. The change in plasma adiponectin levels was significantly correlated with the changes in body mass index (r = -0.5, P = 0.01), waist (r = 0.4, P = 0.04) and hip (r = -0.6, P = 0.0007) circumferences, and steady state plasma glucose levels (r = -0.5, P = 0.04). In multivariate linear regression models, the increase in adiponectin as a dependent variable was significantly related to the decrease in hip circumference (beta = -0.16, P = 0.028), after adjusting body mass index and waist circumference. The change in steady state plasma glucose levels as a dependent variable was related to the increase of adiponectin with a marginal significance (beta = -0.92, P = 0.053), after adjusting body mass index and waist and hip circumferences. In conclusion, body weight reduction increased the plasma levels of a protective adipocytokine, adiponectin. In addition, the increase in plasma adiponectin despite the reduction of the only tissue of its own synthesis suggests that the expression of adiponectin is under feedback inhibition in obesity. PMID- 11502818 TI - Heterozygous mutation in the cholesterol side chain cleavage enzyme (p450scc) gene in a patient with 46,XY sex reversal and adrenal insufficiency. AB - Cytochrome P450scc, the mitochondrial cholesterol side chain cleavage enzyme, is the only enzyme that catalyzes the conversion of cholesterol to pregnenolone and, thus, is required for the biosynthesis of all steroid hormones. Congenital lipoid adrenal hyperplasia is a severe disorder of steroidogenesis in which cholesterol accumulates within steroidogenic cells and the synthesis of all adrenal and gonadal steroids is impaired, hormonally suggesting a disorder in P450scc. However, congenital lipoid adrenal hyperplasia is caused by mutations in the steroidogenic acute regulatory protein StAR; it has been thought that P450scc mutations are incompatible with human term gestation, because P450scc is needed for placental biosynthesis of progesterone, which is required to maintain pregnancy. In studying patients with congenital lipoid adrenal hyperplasia, we identified an individual with normal StAR and SF-1 genes and a heterozygous mutation in P450scc. The mutation was found in multiple cell types, but neither parent carried the mutation, suggesting it arose de novo during meiosis, before fertilization. The patient was atypical for congenital lipoid adrenal hyperplasia, having survived for 4 yr without hormonal replacement before experiencing life-threatening adrenal insufficiency. The P450scc mutation, an in frame insertion of Gly and Asp between Asp271 and Val272, was inserted into a catalytically active fusion protein of the P450scc system (H2N-P450scc Adrenodoxin Reductase-Adrenodoxin-COOH), completely inactivating enzymatic activity. Cotransfection of wild-type and mutant vectors showed that the mutation did not exert a dominant negative effect. Because P450scc is normally a slow and inefficient enzyme, we propose that P450scc haploinsufficiency results in subnormal responses to ACTH, so that recurrent ACTH stimulation leads to a slow accumulation of adrenal cholesterol, eventually causing cellular damage. Thus, although homozygous absence of P450scc should be incompatible with term gestation, haploinsufficiency of P450scc causes a late-onset form of congenital lipoid adrenal hyperplasia that can be explained by the same two-hit model that has been validated for congenital lipoid adrenal hyperplasia caused by StAR deficiency. PMID- 11502819 TI - Mutational analysis of the PRL receptor gene in human breast tumors with differential PRL receptor protein expression. AB - PRL is a major growth and differentiating hormone in the human breast, with activation of the PRL-PRL receptor complex increasingly recognized as an important mechanism in the induction and progression of mammary tumors. Although constitutive activation of various hormone and growth factor receptors is newly recognized as a common cause of tumor development, the PRL receptor gene has not been analyzed for similar aberrations in breast and other tumors. Therefore, using bacterial artificial chromosomes containing the PRL receptor gene and intron-spanning PCR, we determined the exon-surrounding intron sequences providing primers for the first analysis of the entire coding region of the human PRL receptor gene. We examined the presence of PRL receptor in 41 breast tumors by immunohistochemistry and attempted a correlation of its expression to pathological grading of the disease. Then tumor cells were isolated by laser capture microdissection to examine DNA from 30 patients for PRL receptor mutations. The PRL receptor immunoreactive score did not correlate to the tumor size, histopathological grading, age, or family history of patients. PRL receptor immunoreactivity was predominantly found in steroid hormone receptor-positive tumors, but without overall correlation of immunoreactive score. In both PRL receptor-positive and PRL receptor- negative breast cancer cells, direct sequencing of the coding sequence of the PRL receptor gene did not detect any somatic or hereditary gene aberrations. In conclusion, PRL receptor mutations do not appear to be common in human breast cancer, suggesting that constitutive activation of the PRL receptor can be excluded as a major cause of mammary tumor genesis. The molecular structure of the PRL receptor seems to remain intact in tumor tissue, and systemic and local production of PRL may participate in tumor cell growth and proliferation through functional receptors. PMID- 11502820 TI - Estrogen increases endothelial carbon monoxide, heme oxygenase 2, and carbon monoxide-derived cGMP by a receptor-mediated system. AB - Carbon monoxide, a gaseous activator of soluble guanylyl cyclase formed by a subtype of the enzyme heme oxygenase designated heme oxygenase-2 in vascular endothelium, has been found to dilate blood vessels independently from nitric oxide. Because of the parallels between nitric oxide and carbon monoxide, we speculated that estrogen might affect carbon monoxide production in vascular endothelium. Endothelial cells of human origin (umbilical vein and uterine artery) were incubated for 4 or 24 h with 10(-12)-10(-6) M 17beta-estradiol. 17beta-Estradiol, at a concentration such as that attained during the ovulatory phase of the menstrual cycle (10(-10) M), administrated for 4 h led to a 2-fold increase in intracellular carbon monoxide production and heme oxygenase-2 protein levels (P < 0.05). A reporter assay, measuring the formation of cGMP as the direct product of carbon monoxide-induced activation of soluble guanylyl cyclase in endothelial cells, also revealed a 56% increase in cellular cGMP after treatment with 10(-10) M E2 17beta-estradiol (P < 0.05). By contrast, higher 17beta-estradiol concentrations had no significant respective effects due to nitric oxide synthase inhibition of carbon monoxide release. This 17beta estradiol effect appeared to be ER dependent, as preincubation with tamoxifen (10(-6) M) blocked the stimulatory effect of 17beta-estradiol in each instance. Our preliminary data indicate a potential role for carbon monoxide as a biological messenger molecule in estrogen-mediated regulation of vascular tone. PMID- 11502821 TI - X-linked hypophosphatemia attributable to pseudoexons of the PHEX gene. AB - X-linked hypophosphatemia is commonly caused by mutations of the coding region of PHEX (phosphate-regulating gene with homologies to endopeptidases on the X chromosome). However, such PHEX mutations are not detected in approximately one third of X-linked hypophosphatemia patients who may harbor defects in the noncoding or intronic regions. We have therefore investigated 11 unrelated X linked hypophosphatemia patients in whom coding region mutations had been excluded, for intronic mutations that may lead to mRNA splicing abnormalities, by the use of lymphoblastoid RNA and RT-PCRs. One X-linked hypophosphatemia patient was found to have 3 abnormally large transcripts, resulting from 51-bp, 100-bp, and 170-bp insertions, all of which would lead to missense peptides and premature termination codons. The origin of these transcripts was a mutation (g to t) at position +1268 of intron 7, which resulted in the occurrence of a high quality novel donor splice site (ggaagg to gtaagg). Splicing between this novel donor splice site and 3 preexisting, but normally silent, acceptor splice sites within intron 7 resulted in the occurrences of the 3 pseudoexons. This represents the first report of PHEX pseudoexons and reveals further the diversity of genetic abnormalities causing X-linked hypophosphatemia. PMID- 11502822 TI - Abdominal visceral fat and fasting insulin are important predictors of 24-hour GH release independent of age, gender, and other physiological factors. AB - Numerous physiological factors modulate GH secretion, but these variables are not independent of one another. We studied 40 younger (20-29 yr.; 21 men and 19 women) and 62 older (57-80 yr.; 35 men and 27 women) adults to determine the contributions of several demographic and physiological factors to the variability in integrated 24-h GH concentrations. Serum GH was measured every 10 min for 24 h in an enhanced sensitivity chemiluminescence assay. The predictor variables included: age group (young or old), gender, abdominal visceral fat (by computed tomography), total body fat mass and percentage body fat by dual-energy x-ray absorptiometry, serum IGF-I, fasting serum insulin, 24-h mean estradiol and testosterone, and peak oxygen uptake by graded exercise (treadmill) testing. Multiple ordinary least squares regression analysis was used to quantitatively assess the individual contribution that each predictive measure made to explain the variability among values of integrated 24-h GH concentrations while in the presence of the remaining predictors. The model explained 65% of the variance in integrated 24-h GH concentrations. Abdominal visceral fat (P < 0.002) and fasting insulin (P < 0.008) were consistently important predictors of integrated 24-h GH concentrations independent of age group, gender, and all other predictor variables. Although serum IGF-I was an important overall predictor of integrated 24-h GH concentrations (P = 0.002), this relationship was present only in the young subjects and was modulated by gender. The remaining variables failed to contribute significantly to the model. We conclude that abdominal visceral fat and fasting insulin are important predictors of integrated 24-h GH concentrations in healthy adults, independent of age and gender. Serum IGF-I is an important predictor of integrated 24-h GH concentrations in young but not older subjects. Bidirectional feedback between each of these three factors and GH secretion may account for the strong relationships observed. PMID- 11502823 TI - Determinants of the effectiveness of glucagon-like peptide-1 in type 2 diabetes. AB - GLP-1 lowers blood glucose in fasting type 2 diabetic patients. To clarify the relation of the effect of GLP-1 to obesity, blood glucose, beta-cell function, and insulin sensitivity, GLP-1 (1.2 pmol/kg.min) was infused iv for 4-6 h into 50 fasting type 2 diabetic patients with a wide range of age, body mass index, HbA1c, and fasting plasma glucose. The effectiveness of GLP-1 was evaluated by calculation of a glucose disappearance constant for each individual (Kg, linear slope of log-transformed plasma glucose), and by the lowest stable glucose level (Nadir plasma glucose) obtained during the infusion. Grouped according to fasting plasma glucose (<10, 10-15, >15 mmol/liter), Kg values were 0.45 +/- 0.03, 0.38 +/- 0.04, and 0.28 +/- 0.04%/min (P = 0.005), and Nadir plasma glucose values were 4.7 +/- 0.1 (3.9-5.9), 5.8 +/- 0.4 (4.3-8.4), and 8.7 +/- 1.4 (6.2-18.7) mmol/liter (P = 0.0003). Nonresponders were not identified. Multiple regression analysis with Kg or Nadir plasma glucose as the dependent parameter and body mass index, age, gender, diabetes duration, and significantly correlated parameters (in multiple regression for Kg: fasting plasma glucose, fasting nonesterified fatty acid, dipeptidyl peptidase activity, peak insulin, and the logarithm of beta-cell function; and for Nadir plasma glucose: fasting plasma glucose, fasting nonesterified fatty acid, dipeptidyl peptidase activity, delta glucagon decrement, F-GLP-1 total, logarithm of beta-cell function, and Kg) as independent parameters resulted in fasting plasma glucose as the only significant predictor of Kg, and fasting plasma glucose and Kg as predictors of Nadir plasma glucose. Kg and Nadir plasma glucose were neither influenced by treatment nor by neuropathy per se. In conclusion, GLP-1 lowers plasma glucose in type 2 diabetes regardless of severity, but glucose elimination is faster and obtained glycemic level lower in patients with the lower fasting plasma glucose. Not all patients can be expected to reach normoglycemia. PMID- 11502824 TI - No association of an interleukin 4 gene promoter polymorphism with Graves' disease in the United Kingdom. AB - Graves' disease (GD) is an autoimmune thyroid disease of unknown etiology, although predisposition to the development of this disease is thought to be caused by both genetic and environmental factors. Recently, an association between a promoter polymorphism of the interleukin 4 gene and GD has been reported. A C-T base change at position -590 showed modest protection against the development of GD in a United Kingdom data set of 135 patients with GD and 101 controls. This polymorphism was, therefore, investigated in a much larger case control cohort of 384 patients with GD and 288 control subjects using PCR, followed by restriction fragment length polymorphism analysis. No protective effect of the T allele of this polymorphism was observed in our data set, and indeed no significant difference in either allelic or genotypic distribution was seen between the patient and control groups. Moreover, calculation of probabilities indicate that the original study lacked sufficient power to support the conclusions drawn. Our data support the hypothesis that the C-T promoter polymorphism of the interleukin 4 gene does not confer protection against the development of GD in Caucasians in the United Kingdom. PMID- 11502825 TI - Cerebral blood flow and glucose metabolism in hypothyroidism: a positron emission tomography study. AB - Hypothyroidism is often associated with defective memory, psychomotor slowing, and depression. However, the relationship between thyroid status and cognitive or psychiatric disturbances remains unclear. Using psychometric scales, 10 patients who had undergone total thyroidectomy for thyroid carcinoma were evaluated for depression, anxiety, and psychomotor slowing; they were examined both when euthyroid and hypothyroid after thyroid hormone withdrawal. Positron emission tomography was used, with oxygen-15-labeled water and fluorine-18F-labeled 2 deoxy-2fluoro-D-glucose as the tracers, to correlate the regional cerebral blood flow and cerebral glucose metabolism with the mental state in patients. Two different image analysis techniques (regions of interest and statistical parametric maps) were applied. In hypothyroidism, there was a generalized decrease in regional cerebral blood flow (23.4%, P < 0.001) and in cerebral glucose metabolism (12.1%, P < 0.001) and there were no specific local defects. Patients were also significantly more depressed (P < 0.001), anxious (P < 0.001) and psychomotor slowed (P < 0.005) in hypo than in euthyroid status. These results indicate that the brain activity was globally reduced in severe hypothyroidism of short duration without the regional modifications usually observed in primary depression. PMID- 11502826 TI - Increased vascular endothelial growth factor serum concentrations may help to identify patients with onset of type 1 diabetes during childhood at risk for developing persistent microalbuminuria. AB - This study was designed to evaluate whether vascular endothelial growth factor serum concentrations may identify adolescents with onset of type 1 diabetes during childhood at greater risk to develop persistent microalbuminuria and incipient diabetic nephropathy. In January 1989, vascular endothelial growth factor serum levels were measured in 101 normoalbuminuric diabetic children and adolescents (aged 7-14.9 yr; onset of diabetes before age 18 yr; duration of diabetes >7 yr). Participants were clinically examined at baseline and annually thereafter. Vascular endothelial growth factor serum concentrations were measured every year during the 8-yr follow-up period. Over 8 yr, 11 of 101 patients (10.9%) developed persistent microalbuminuria; no patient developed overt nephropathy. The risk of developing microalbuminuria was higher in children with increased vascular endothelial growth factor serum levels (using 160 pg/ml as the arbitrary cut-off point; group 1) compared with those with normal vascular endothelial growth factor serum levels at the beginning of the study (group 2; 19.2 vs. 2.0%; P < 0.01; sensitivity, 90.9%; specificity, 53.3%). The odds ratio for the occurrence of microalbuminuria after adjustment for confounding variables (albumin excretion rate, sex, hemoglobin A(1c), mean blood pressure, cholesterol, and triglycerides) in type 1 diabetic adolescents with elevated vascular endothelial growth factor serum levels was 4.1 (95% confidence interval, 2.0 10.9). These results suggest that vascular endothelial growth factor serum concentrations may be one of the predictors and risk factors for microalbuminuria and incipient diabetic nephropathy in adolescents and young adults with onset of diabetes during childhood. Persistently increased vascular endothelial growth factor serum levels may help to identify normotensive, normoalbuminuric patients with type 1 diabetes who are predisposed to develop persistent microalbuminuria later in life. PMID- 11502827 TI - Isolated GH deficiency with dominant inheritance: new mutations, new insights. AB - Autosomal dominantly inherited isolated GH deficiency is caused by mutations of GH-1 that alter the normal structure of GH. We studied 16 familial cases and 1 sporadic case with isolated GH deficiency type II from 1 Dutch and 4 German families by direct sequencing of PCR-amplified genomic DNA and ectopic transcript analysis of lymphocyte mRNA. In addition, the clinical data of the affected individuals were analyzed. Two previously reported mutations and 1 novel splice site mutation in intron III of GH-1 (+1G to C and +1G to A; new, +2T to C) were detected that cause exon 3 skipping. We also discovered a novel G6191 to T missense mutation in exon 4 of GH-1 that changes valine 110, which is highly conserved in mammalian and several nonmammalian GH, to phenylalanine. Splicing of the primary RNA transcript was not affected by this mutation, which is very likely to alter the normal GH structure at the protein level. The onset of growth failure was earlier, and the degree was more severe in affected children with GH 1 splice site mutations compared with those in children with the GH-1 missense mutation. In addition, the severity of the phenotype was variable, even within the same family. The age at diagnosis was between 0.8-9.6 yr (median, 5.1 yr); height at diagnosis was between -2.5 and -8.1 SD score (median, -4.0). Most of the children were lean at diagnosis, with a body mass index ranging from -1.7 to +3.3 SD score (median, -0.5). The 5 affected adults had final heights between 1.8 and -4.5 SD score (median, -3.6), centripetal obesity, and muscular hypotrophy. Before therapy, IGF-I and IGF-binding protein-3 serum levels of all affected children were severely diminished (<<5th percentiles for age). The maximum GH peak in a total of 25 stimulation tests was between 0.1-5.0 microg/liter (median, 0.9), indicating severe GH deficiency. The height of the adenohypophysis studied by magnetic resonance imaging was normal in 2 affected children and mildly decreased in 2 others. Substitution with GH resulted in good catch-up growth in all treated children. Children with severe GH and IGF-I deficiencies, but normal size of the adenohypophysis should be examined for GH-1 splice site and missense mutations. The observed discrepancy between the very homogeneous hormone data proving severe GH and IGF-I deficiencies and the high variability of growth failure even within the same family suggests that the onset and predominance of GH-dependent growth during infancy are individually different and modified by as yet unknown factors. PMID- 11502828 TI - Abnormal GH receptor signaling in children with idiopathic short stature. AB - Peripheral GH insensitivity may underlie idiopathic short stature in children. As the clinical and biochemical hallmarks of partial GH insensitivity have not yet been clearly elucidated, the identification of such patients is still difficult. We integrated functional, biochemical, and molecular studies to define the more reliable marker(s) of GH insensitivity. In particular, we measured GH receptor transducing properties through GH-induced protein tyrosine phosphorylation in patients' peripheral blood mononuclear cells and performed direct sequencing analysis of GH receptor-coding exons. Five of 14 idiopathic short stature patients with low basal IGF-I levels showed low or absent IGF-I increment after 4 d of GH administration. However, a prolonged GH stimulation induced in 3 of them an increase in IGF-I 40% above the baseline value. The IGF-binding protein-3 behavior paralleled that of IGF-I. The 2 GH-unresponsive subjects showed an abnormal tyrosine phosphorylation pattern after GH challenge. Sequence analysis of the GH receptor gene revealed a heterozygous mutation resulting in an Arg to Cys change (R161C) in exon 6 in only 1 patient, who had normal GH receptor responsiveness. Our findings indicate that abnormal GH receptor signaling may underlie idiopathic short stature even in the absence of GH receptor mutations. Thus, combining the 4-d IGF-I generation test and the analysis of GH-induced protein tyrosine phosphorylation is a useful tool to help identify idiopathic short stature patients with partial GH insensitivity. PMID- 11502829 TI - Mutational analysis and genotype-phenotype correlation of the PHEX gene in X linked hypophosphatemic rickets. AB - PHEX is the gene defective in X-linked hypophosphatemic rickets. In this study, analysis of PHEX revealed mutations in 22 hypophosphatemic rickets patients, including 16 of 28 patients in whom all 22 PHEX exons were studied. In 13 patients, in whom no PHEX mutation had been previously detected in 17 exons, the remaining 5 PHEX exons were analyzed and mutations found in 6 patients. Twenty different mutations were identified, including 16 mutations predicted to truncate PHEX and 4 missense mutations. Phenotype analysis was performed on 31 hypophosphatemic rickets patients with PHEX mutations, including the 22 patients identified in this study, 9 patients previously identified, and affected family members. No correlation was found between the severity of disease and the type or location of the mutation. However, among patients with a family history of hypophosphatemic rickets, there was a trend toward more severe skeletal disease in patients with truncating mutations. Family members in more recent generations had a milder phenotype. Postpubertal males had a more severe dental phenotype. In conclusion, although identifying mutations in PHEX may have limited prognostic value, genetic testing may be useful for the early identification and treatment of affected individuals. Furthermore, this study suggests that other genes and environmental factors affect the severity of hypophosphatemic rickets. PMID- 11502830 TI - Hypopituitarsim is associated with triglyceride enrichment of very low-density lipoprotein. AB - The dyslipidemia associated with hypopituitarism may contribute to increased vascular mortality. The atherogenic potential of lipoproteins is determined not only by concentration but also by their composition. We therefore studied very low-density lipoprotein composition and apolipoprotein B kinetics in 16 hypopituitary subjects and 16 controls. Hypopituitarism was associated with reduced high-density lipoprotein cholesterol (0.98[0.82-1.18] vs. 1.35[1.15-1.41] mmol/liter, P < 0.001) and increased triglyceride concentrations (1.64[1.09-2.77] vs. 1.12[0.66-1.67] mmol/liter, P = 0.01). Total (P = 0.76) and low-density lipoprotein cholesterol (P = 0.56) concentrations were similar. Very low-density lipoprotein- triglyceride was significantly increased (1.48[1.02-2.55] vs. 0.9[0.31-2.30] mmol/liter, P = 0.004), but very low-density lipoprotein cholesterol levels were similar (P = 0.93). The molar ratios of very low-density lipoprotein-triglyceride:apolipoprotein B (6193[4283-9566] vs. 3599[3188-6854], P = 0.005) and very low-density lipoprotein-triglyceride:cholesterol (2.8[1.98 3.78] vs. 1.6[1.44-2.80], P < 0.003) were significantly increased; very low density lipoprotein-cholesterol:apolipoprotein B molar ratios (P = 0.93) were similar. Very low-density lipoprotein apolipoprotein B fractional synthetic rate (a measure of apolipoprotein B catabolism, P = 0.42) and pool size (P = 0.63) were similar. The very low-density lipoprotein apolipoprotein B absolute synthetic rate (a measure of apolipoprotein B synthesis) tended to be higher in hypopituitarism (17.7[2.91-19.50] vs. 26.6[19.64-28.05] mg/kg per day, P = 0.24) but failed to reach statistical significance. The absolute synthetic rate, and hence very low-density lipoprotein production, correlated with very low-density lipoprotein triglyceride:apolipoprotein B ratio (P = 0.02, Rs = 0.63), suggesting that triglyceride enrichment of very low-density lipoprotein is important in the mechanism underlying very low-density lipoprotein overproduction in hypopituitarism. Because triglyceride-enriched lipoproteins are proatherogenic, this may contribute to the vascular mortality observed in hypopituitarism. The reasons for these observations are unknown; GH deficiency or routine endocrine replacement may be important. PMID- 11502831 TI - Two Chinese families with Pendred's syndrome--radiological imaging of the ear and molecular analysis of the pendrin gene. AB - We report two families in whom the index cases satisfied the classical diagnostic criteria of Pendred's syndrome. In family I, two siblings were deaf, and one was normal. In family II, both parents and two offspring were deaf. Computed tomography scans performed in five of six of these deaf individuals showed enlarged vestibular aqueducts in all cases, and Mondini cochlea only in family II. Affected members in family I were compound heterozygotes inheriting the paternal allele with a novel mutation S398del in exon 10 and a maternal allele with two mutations IVS13+9C-->G in intron 13, in addition to H723R. In family II, the mother and one child carried both the novel intronic IVS8-2A-->G and H723R mutations, whereas the father and index case were homozygous for the IVS8-2A-->G mutation. A perchlorate discharge test was positive in 50% of cases tested. In conclusion, we concur that radiological and molecular studies should be performed for confirmation of Pendred's syndrome. We report, for the first time, a Pendred's syndrome family in which affected members had three mutations, as well as a second family in whom the intermarriage of two Pendred's syndrome patients resulted in Pendred's syndrome offspring. PMID- 11502832 TI - A prospective, randomized study of endometrial telomerase during the menstrual cycle. AB - The purpose of this study was to characterize telomerase activity during the menstrual cycle, focusing on the luteal phase. A total of 84 endometrial biopsy samples were obtained from 72 participants. Daily urinary LH testing (OvuQuick, Quidel) was used to establish the day of the LH rise, and participants were randomized to return during the secretory phase. Twelve women returned on the identical day during the luteal phase of a subsequent cycle to allow intercycle comparisons of telomerase activity. Telomerase activity was evaluated using a modified TRAP-eze (Intergen) detection protocol. At the time of each endometrial biopsy, serum estrogen and progesterone were measured. Proliferative phase endometrium showed high telomerase activity. At the onset of the luteal phase telomerase activity was high, but it decreased during the early luteal phase, disappeared by the midluteal phase (6 d after LH surge detected), and then rose to moderate levels in the late luteal phase beginning on luteal d 10. Serum progesterone levels were inversely related to telomerase activity. In conclusion, endometrial telomerase activity is dynamic: high during the proliferative phase but inhibited during the midsecretory phase of the menstrual cycle. The timing of expression coincides with the rise and fall of progesterone levels and the time period of maximal uterine receptivity for embryo implantation. This supports a relationship between sex steroid levels and telomerase regulation. PMID- 11502833 TI - Effects of aging and estradiol supplementation on GH axis dynamics in women. AB - GH and IGF-I secretion decrease with age. The decline in serum GH with age appears to be associated with menopause. Prior studies of GH release before and after oral and transdermal hormonal replacement in the postmenopausal patient have shown no change or an increase in GH secretion. To distinguish the somatotropic axis effects of aging from those of estrogen deficiency, we compared eight prematurely menopausal women, aged 25-40 yr, with eight postmenopausal women, aged 51-70 yr, both before and after estradiol replacement. All women had a body mass index below 28 kg/m2. All were evaluated twice with frequent blood sampling every 10 min for 24 h. Studies were performed in the absence of exogenous hormones and 6-8 wk after transdermal estradiol replacement, targeted to achieve a serum estradiol level of 367 pmol/liter. GH pulsatility was analyzed. Variables tested included mean GH levels, interpulse baseline mean, pulse frequency per 24 h, and pulse amplitude. Transdermal estrogen replacement had a significant effect on mean GH levels and mean basal GH levels in both the premature ovarian failure and the age-appropriate postmenopausal group. No differences were noted in GH pulse frequency, GH pulse amplitude, IGF-I, IGF binding protein-1, and IGF-binding protein-3 before and after treatment. A pronounced age effect was noted between the two groups. The premature ovarian failure women secreted significantly greater mean GH than the age-appropriate postmenopausal group regardless of treatment, with a significance level of P = 0.026. Interpulse baseline GH means were greater in the premature ovarian failure women than in the age-appropriate postmenopausal group, but the significance of this relationship was obliterated after adjustment for body mass index. Pulse amplitude was significantly increased in the premature ovarian failure women compared with age-appropriate postmenopausal women (P = 0.006). No significant changes were detected in the GH pulse frequency between the premature ovarian failure and postmenopausal groups. We conclude that moderate doses of transdermal estradiol supplementation do not exert a great effect on the somatotropic axis in women. Age and body composition appear to be the predominant influences on GH activity in women. PMID- 11502834 TI - Expression of scavenger receptor class B1 in endometrium and endometriosis. AB - Endometriosis is characterized by the presence of endometrial glands and stroma within the peritoneum and other extrauterine sites. The presence of aromatase, a key enzyme in the biosynthesis of estradiol, has been demonstrated in eutopic endometrial samples of women with moderate to severe endometriosis, but not in those of disease-free women. Animal studies have shown that high density lipoprotein provides precursor cholesterol pool for steroidogenesis. Scavenger receptor class B1, a 82-kDa cell surface protein, is involved in the binding of high density lipoprotein to target cells and promotes cholesteryl ester uptake. In this study we detected the presence of scavenger receptor class B1 in eutopic and ectopic endometrium. There was more scavenger receptor class B1 protein associated with endometriosis compared with matched endometrium. Two bands (82 and 45 kDa) were detected in the endometrium and endometriosis samples. Glycanase treatment indicated that the 45-kDa protein might be a nonglycosylated form of scavenger receptor class B1. Immunostaining of fixed tissues detected scavenger receptor class B1 in glandular epithelium of both tissues. Scavenger receptor class B1 and aromatase mRNA were increased in endometriosis tissues. Scavenger receptor class B1 expression in the endometrium and endometriosis supports a role for this receptor in the uptake of high density lipoprotein cholesterol, possibly supporting in situ estrogen production, which is detrimental in the progression of endometriosis. PMID- 11502835 TI - Altered expression of novH is associated with human adrenocortical tumorigenesis. AB - NOVH belongs to the CCN (CTGF/CYR61/NOV) family of proteins, some of which have chemotactic, mitogenic, adhesive, and angiogenic properties. Whereas ctgf and cyr61 are growth factor-inducible, immediate-early genes, nov is expressed in growth-arrested or quiescent cells. As nov expression has been shown to be altered in both avian and human nephroblastomas and to be a target of WT1 regulation, NOV may play important roles in normal nephrogenesis and the development of Wilms' tumors. The aim of this study was to determine whether changes in novH expression were associated with tumorigenesis in tissues other than those of the kidney. We showed by Northern blotting and immunohistochemistry that among human adult endocrine tissues, the adrenal gland is a major site of novH expression, and that in adult and fetal adrenal tissue, novH is primarily expressed in the adrenal cortex. Studies with 12 benign and 18 malignant adrenocortical tumors revealed that the levels of novH mRNA and protein decreased significantly (P < 0.004) with progression of adrenocortical tumors from a benign to a malignant state. Although the localization of NOVH did not change, the N glycosylation profile of benign and malignant tumors differed considerably from that of normal adrenocortical tissue, and these differences may affect the biochemical properties of the molecule. The properties of NOVH here provide the first evidence that this member of the CCN family could be involved in adrenocortical tumor development. PMID- 11502836 TI - Autosomal dominant GH deficiency due to an Arg183His GH-1 gene mutation: clinical and molecular evidence of impaired regulated GH secretion. AB - G to A transition at position 6664 of the GH-1 gene results in the substitution of Arg183 by His (R183H) in human GH protein and causes a new form of autosomal dominant isolated GH deficiency (type II). Although a weak GH release after standard pharmacological provocation tests is observed in these affected individuals, the dominant inheritance pattern is postulated to be caused by a blockade of the GH-regulated secretion in the somatotrophs. The aim of this study was to analyze the impact of this autosomal dominant mutation not only at a clinical, but also at a cellular, level. The results of the different stimulation tests showed first that the patient possesses a severely impaired, but releasable, GH store, and second that the GH secretion is blocked in a time dependent and reversible way. To confirm these clinical data, cell culture studies were performed looking at the regulated secretory pathway of GH using AtT 20 cells. Importantly, we were able to show that when the R183H mutant GH was expressed in AtT-20 cells, secretagogue (forskolin) induced a normal R183H GH regulated secretion, but in AtT-20 cells coexpressing both the R183H mutant GH and the normal GH, forskolin-induced GH secretion was markedly reduced. Together, the experiments seem to support the hypothesis that R183H mutant GH severely impaired the GH-regulated secretion and may, therefore, be the cause of this specific form of isolated GH deficiency type II. PMID- 11502837 TI - Germline RET 634 mutation positive MEN 2A-related C-cell hyperplasias have genetic features consistent with intraepithelial neoplasia. AB - C-cell hyperplasias are normally multifocal in multiple endocrine neoplasia type 2A. We compared clonality, microsatellite pattern of tumor suppressor genes, and cellular kinetics of C-cell hyperplasia foci in each thyroid lobe. We selected 11 females from multiple endocrine neoplasia type 2A kindred treated with thyroidectomy due to hypercalcitoninemia. C-cell hyperplasia foci were microdissected for DNA extraction to analyze the methylation pattern of androgen receptor alleles and microsatellite regions (TP53, RB1, WT1, and NF1). Consecutive sections were selected for MIB-1, pRB1, p53, Mdm-2, and p21WAF1 immunostaining, DNA content analysis, and in situ end labeling. Appropriate tissue controls were run. Only two patients had medullary thyroid carcinoma foci. Nine informative C-cell hyperplasia patients showed germline point mutation in RET, eight of them with the same androgen receptor allele preferentially methylated in both lobes. C-cell hyperplasia foci showed heterogeneous DNA deletions revealed by loss of heterozygosity of TP53 (12 of 20), RB1 (6 of 14), and WT1 (4 of 20) and hypodiploid G0/G1 cells (14 of 20), low cellular turnover (MIB-1 index 4.5%, in situ end labeling index 0.03%), and significantly high nuclear area to DNA index ratio. MEN 2A (germline point mutation in RET codon 634) C-cell hyperplasias are monoclonal and genetically heterogeneous and show down-regulated apoptosis, findings consistent with an intraepithelial neoplasia. Concordant X-chromosome inactivation and interstitial gene deletions suggest clone expansions of precursors occurring at a point in embryonic development before divergence of each thyroid lobe and may represent a paradigm for other germline mutations. PMID- 11502838 TI - Expression of adrenomedullin by human placental cytotrophoblasts and choriocarcinoma JAr cells. AB - Adrenomedullin is a multifunctional peptide expressed in a variety of tissues. This study was conducted to investigate the expression of adrenomedullin and its mRNA by human trophoblasts and the possible existence of adrenomedullin receptor in those cells. Human placentas in all three trimesters were obtained from patients undergoing therapeutic abortions and deliveries. Total RNA was extracted from placental trophoblastic tissues and JAr choriocarcinoma cells, and the expression of adrenomedullin mRNA was determined by RT-PCR. Immunohistochemical analysis was performed by the avidin/biotin immunoperoxidase method using a specific antibody to adrenomedullin. The secretion of adrenomedullin by JAr cells cultured in medium containing [35S]cysteine-[35S]methionine was determined by immunoprecipitation followed by PAGE. The presence of adrenomedullin receptor in JAr cells was examined using a binding assay with [125I]rat adrenomedullin. Adrenomedullin mRNA was expressed by human placental trophoblastic tissues in all three trimesters and by JAr cells. Immunohistochemical analysis revealed that adrenomedullin is expressed by cytotrophoblasts in placentas in all three trimesters, but not by syncytiotrophoblasts. The expression of adrenomedullin in the cytotrophoblast was most abundant in first trimester placenta and became less abundant during the course of pregnancy. JAr cells synthesized and secreted immunoreactive adrenomedullin. Binding assay with [125I]rat adrenomedullin demonstrated specific binding of adrenomedullin to JAr cells, indicating the existence of a specific receptor for adrenomedullin in trophoblastic cells. Adrenomedullin is transcribed and secreted by cytotrophoblastic cells that possess adrenomedullin receptor. Adrenomedullin may play a potential role as an autocrine/paracrine factor in the growth of cytotrophoblasts, especially in early gestation. PMID- 11502839 TI - A novel mutation (Q40P) in PAX8 associated with congenital hypothyroidism and thyroid hypoplasia: evidence for phenotypic variability in mother and child. AB - Congenital hypothyroidism associated with thyroid hypoplasia can be caused by several genetic defects, including mutations in the TSHbeta-subunit, the TSH receptor, the G(s)alpha-subunit, and the transcription factor PAX8. Four girls with sporadic congenital hypothyroidism and hypoplastic thyroid glands were analyzed for mutations in PAX8 and TTF2 (FKHL15). Mutations in the coding region of the TSHbeta-subunit gene, the TSH receptor gene, and exons 8 and 9 of G(s)alpha had been excluded previously. Serum TSH concentrations were 150 mU/liter or more, TG levels were within normal limits, and thyroid autoantibodies were absent. Technetium scintigraphies did not reveal the presence of thyroid tissue, but ultrasonography documented hypoplastic, normally located glands. One patient was found to harbor a heterozygous transversion 119A-->C in exon 3 of PAX8 replacing a conserved glutamine by proline in the paired box domain (Q40P). Analysis of her family members revealed that her mother, who has a thyroid gland of normal size and mild, adult-onset autoimmune hypothyroidism, is also heterozygous for this mutation. Functional analyses of the PAX8 Q40P mutation showed impaired binding to a PAX8 response element and absent trans-activation of a thyroid peroxidase promoter luciferase reporter gene. These findings confirm the important role of PAX8 in the development of the thyroid, but they indicate that PAX8 gene mutations may have a variable penetrance or expressivity. The absence of mutations in the coding sequences of the analyzed genes in the three other patients supports the concept that the pathogenesis of congenital hypothyroidism associated with thyroid hypoplasia is diverse. PMID- 11502840 TI - Overexpression of CXC chemokines by an adrenocortical carcinoma: a novel clinical syndrome. AB - A patient with adrenocortical carcinoma presented with fever, leukocytosis, and increased acute phase reactants. The tumor was infiltrated with neutrophils. Immunohistochemical staining of the tumor showed positive signal for epithelial neutrophil-activating protein-78, an angiogenic and chemotactic CXC chemokine. Conditioned medium from tumor-derived cells (RL-251) showed high concentration of IL-8, epithelial neutrophil-activating protein-78, Gro alpha, and Gro gamma, angiogenic CXC chemokines with a potential role in tumorigenesis. An adrenal cancer/severe combined immunodeficiency mouse chimera was developed. Mice grew tumors rapidly, and circulating levels of IL-8 and epithelial neutrophil activating protein-78 were detected. In contrast, animals transplanted with NCI H295 cells, a nonchemokine-secreting cell line, grew tumors more slowly and did not have detectable chemokine levels. Similar to the patient, mice with RL-251 tumors developed marked leukocytosis and neutrophilia, and their tumors were infiltrated with neutrophils. Mice were passively immunized with epithelial neutrophil-activating protein-78 antisera. A marked decrease in tumor growth was observed. Potential for chemokine production by other adrenocortical tumors was investigated by RT-PCR in archival material. Six of seven adrenal carcinomas and one of three adenomas had cDNA for IL-8; six of seven carcinomas and the three adenomas had cDNA for epithelial neutrophil-activating protein-78. We concluded that the clinical presentation of this case resulted from increased tumor production of chemotactic chemokines. Through their angiogenic and chemotactic properties these chemokines may play an important role in adrenal tumorigenesis. PMID- 11502841 TI - nm23-H1 immunoreactivity as a prognostic factor in differentiated thyroid carcinoma. AB - Several prognostic factors have been proposed to identify the patients at risk to develop metastases in differentiated thyroid carcinoma. Reduced nm23-H1 expression (a metastatic suppressor gene) has been correlated with high tumor metastatic potential in various human carcinomas, but the results obtained in differentiated thyroid carcinoma remain controversial. To elucidate the usefulness of nm23-H1 as a differentiated thyroid carcinoma prognosis factor, we evaluate the relationship between nm23-H1 immunoreactivity as well as both clinical status and patient outcome. For this purpose, thyroid resected specimens obtained from 94 differentiated thyroid carcinoma consecutive patients (64 papillary and 30 follicular) with at least 5 yr of follow-up were stained using monoclonal antibody to nm23-H1. We did not observe any relationship between nm23 H1 immunoreactivity and age, gender, initial differentiated thyroid carcinoma stage, local recurrence, or distant metastases in patients with papillary carcinoma. However, in patients with follicular carcinoma, a significant inverse association between metastatic disease and the expression of nm23-H1 product was obtained (P < 0.05). In addition, significant differences were found in the survival curves according to nm23-H1 immunoreactivity (log-rank P < 0.01). Finally, nm-23-H1 immunoreactivity was more specific but less sensitive than AMES score to predict metastases. In conclusion, our results suggest that nm23-H1 immunostaining could be added to the classic prognostic factors currently used to predict the outcome of patients with follicular thyroid carcinoma. PMID- 11502842 TI - Reorganization of myofilament proteins and decreased cGMP-dependent protein kinase in the human uterus during pregnancy. AB - Excessive or premature contractions of uterine smooth muscle may contribute to preterm labor. Contractile stimuli induce myosin and actin filament interactions through calcium-dependent myosin phosphorylation. The mechanisms that maintain myometrial quiescence until term are not well established, but may include control of calcium levels by nitric oxide and cGMP signaling and thin filament (caldesmon and calponin) regulation. Previously, we reported that myometrial tissues from pregnant rats are not responsive to cGMP due to decreases in cGMP dependent protein kinase. Considering the well documented differences in the endocrinology of parturition among species, this study was conducted to test the hypothesis that the levels and subcellular distribution of caldesmon, calponin, and cGMP-dependent protein kinase are regulated with the hormonal milieu of human pregnancy. Whereas cGMP-dependent protein kinase was significantly reduced in the human uterus during pregnancy, caldesmon expression was significantly increased, and both caldesmon and calponin were redistributed to a readily extractable subcellular pool. These data suggest that cGMP-dependent protein kinase does not mediate gestational quiescence. Redistribution of thin filament-associated proteins, however, may alter uterine smooth muscle tone or the cytoskeletal framework of myocytes to maintain gestation despite the substantial distention that accompanies all intrauterine pregnancies. PMID- 11502843 TI - Absence of constitutively activating mutations in the GHRH receptor in GH producing pituitary tumors. AB - The molecular events leading to the development of GH-producing pituitary tumors remain largely unknown. We hypothesized that activating mutations of the GHRH receptor might occur in a subset of GH-producing pituitary tumors. Genomic DNA samples from 54 GH-producing pituitary tumor tissues were screened for mutations of the GHRH receptor. Eleven homozygous or heterozygous nucleotide substitutions [169G > A (A57T), 338C > T (P113L), 363G > T (E121D), 409C > T (H137Y), 547G > A (D183N), 673G > A (V225I), 749G > A (W250X), 760G > A (V254M), 785G > A (S262N), 880G > A (G294R), 1268G > A (C423Y)] were found in 12 patients (22.2%). The 169G > A substitution (A57T) appears to be a polymorphism (4 patients, 7.4%). E121D and V225I were each found in 2 patients. In 1 patient with the V225I sequence, the substitution was not found in genomic DNA from peripheral leukocytes, suggesting a somatic mutation. A patient with a heterozygous W250X mutation was homozygous for the C423Y substitution. These variant GHRH receptors were studied in transfected TSA-201 cells to evaluate the functional consequences of the amino acid changes. None of the GHRH receptor variants was associated with basal elevation of intracellular cAMP. GHRH induced variable cAMP responses. With the W250X and G294R variants, there was no cAMP stimulation by GHRH, indicating that the mutations are inactivating. Expression of the W250X GHRH receptor on the cell membrane was severely decreased and GHRH binding to the G294R GHRH receptor was impaired. Although GHRH receptor variants are common in GH- producing pituitary adenomas, constitutively activating mutations, as a mechanism for GH-producing pituitary tumors appear to be rare. PMID- 11502844 TI - Mutations in the preproghrelin/ghrelin gene associated with obesity in humans. AB - Ghrelin and preproghrelin sequences were determined in 96 unrelated female subjects with severe obesity (mean body mass index (BMI) 42.3 +/- 3.4 kg/m(2)) and in 96 non-obese female controls (mean BMI 23.0 +/- 1.4 (kg/m2) of the Swedish Obese Subjects cohort. A mutation at amino acid position 51 (Arg51Gln) of the preproghrelin sequence that corresponds to the last amino acid in mature ghrelin product was identified in six (all heterozygotes) obese subjects (6.3%) but not among controls (p < 0.05). The self-reported weight at 20, 30, and 40 years of age tended to be 7.5, 4.7 and 6.4 kg lower, respectively, among obese Gln allele carriers versus obese non-carriers. In addition, a mutation at codon 72 of the preproghrelin gene (Leu72Met) was detected in 15 obese (12 hetero- and 3 homozygotes) and 12 control (all heterozygotes) subjects. This mutation outside the coding region of the mature ghrelin product tended to be associated with lower age of self-reported onset of obesity (15.6 +/- 7.9 vs. 20.5 +/- 10.5 years; p = 0.09). In addition to these two mutations in coding regions, a G274A base change in a non-coding region between exons one and two was found only in two obese individuals. The Arg51Gln amino acid substitution may alter the cleavage site of endoproteases and the length of the mature ghrelin product. The functional significance of the Leu72Met mutation and a G274A base change remains to be determined. In conclusion, the data provide evidence that a low frequency sequence variation in the ghrelin gene could play a role in the etiology of obesity. PMID- 11502845 TI - The role of human chorionic gonadotropin and cAMP/protein kinase A signaling in the differentiation of human endometrial stromal cells. PMID- 11502846 TI - A new century of international telemedicine for radiation-exposed victims in the world. PMID- 11502847 TI - Bone density regression to the mean and the individual patient. PMID- 11502849 TI - Prevalence of primary aldosteronism in unselected hypertensive populations: screening and definitive diagnosis. PMID- 11502852 TI - Distinguishing unilateral aldosteronoma from bilateral disease. PMID- 11502857 TI - Autoimmune melanocyte destruction in vitiligo. PMID- 11502858 TI - Immunological phenotyping of fibroblast cultures from patients with a mitochondrial respiratory chain deficit. AB - Conventional approaches to the diagnosis of mitochondrial respiratory chain diseases, using enzyme assays and histochemistry, are laborious and give limited information concerning the genetic basis of a deficiency. We have evaluated the diagnostic value of 12 monoclonal antibodies to subunits of the four respiratory chain enzyme complexes and F(1)F(0)-ATP synthase. Antibodies were used in immunological studies with skin fibroblast cultures derived from patients with diverse mitochondrial diseases, including patients in which the disease was caused by a nuclear genetic defect and patients known to harbor a heteroplasmic mutation in a mitochondrial tRNA gene. Immunoblotting experiments permitted the identification of specific enzyme assembly deficits and immunocytochemical studies provided clues regarding the genetic origin of the disease. The immunological findings were in agreement with the biochemical and genetic data of the patients. Our study demonstrates that characterization of the fibroblast cultures with the monoclonal antibodies provides a convenient technique to complement biochemical assays and histochemistry in the diagnosis of mitochondrial respiratory chain disorders. PMID- 11502859 TI - Multiplex detection of hotspot mutations by rolling circle-enabled universal microarrays. AB - Detection of somatic low abundance mutations in early cancer development requires a discriminatory, specific, and high-throughput methodology. In this study we report specific, discriminatory detection of low abundance mutations through a novel combination of rolling circle amplification (Nat Genet 1998; 19:225-232) and PCR ligation detection reaction on a universal oligonucleotide microarray (J Mol Biol 1999; 292:251-262). After mutation-specific multiplex ligation and hybridization of 17 pairs of probes to a generic microarray, the ligated probes were visualized. The multiplex mutation-specific ligation is possible only because rolling circle amplification permits quantification of previously undetectable hybridization events conducive to the detection of a single mutation from within a pool of over 100 wild-type alleles. This system is readily adaptable to high-throughput automation using a robot such as the Biomek platform. PMID- 11502860 TI - Rottlerin-independent attenuation of pervanadate-induced tyrosine phosphorylation events by protein kinase C-delta in hemopoietic cells. AB - The understanding and control of many pathophysiological conditions is based on knowledge of subtly regulated intracellular signaling networks. We have found that in pervanadate (PV)-treated J558L myeloma cells, amongst other signaling proteins, protein kinase C (PKC)-delta and src homology 2-containing inositol phosphatase (SHIP) are tyrosine phosphorylated on expression of the B cell receptor, suggesting a role for these proteins in the preformed B cell receptor transducer complex. Rottlerin, a widely used PKC-delta-specific inhibitor, efficiently blocks these PV-induced tyrosine phosphorylation events. Furthermore, PV treatment of bone marrow-derived mast cells (BMMC) also results in tyrosine phosphorylation of PKC-delta, SHIP, and additional proteins. Rottlerin also inhibits these responses, indicating that PKC-delta might play an important enhancing role in the propagation of phosphotyrosine signals in B cells and mast cells and hence in the regulation of function of both cell types. Therefore, BMMC from PKC-delta -/- mice were generated by in vitro differentiation and assayed for tyrosine phosphorylation events in response to PV. Intriguingly, and opposite to the Rottlerin data, PKC-delta -/- BMMC show a stronger response to PV than wild-type cells, suggesting an attenuating role for PKC-delta. This response can be inhibited equally well by Rottlerin, indicating clearly that Rottlerin is not specific for PKC-delta in vivo. A comparison between Rottlerin and the panspecific PKC inhibitor bisindolylmaleimide suggests that Rottlerin also targets kinases beyond the PKC family. Moreover, Ser473 phosphorylation of protein kinase B (PKB) after PV treatment is blocked by Rottlerin as efficiently as by the phosphatidylinositol 3-kinase inhibitor LY294002. In this report, we provide evidence that PKC-delta constitutes a crucial attenuating factor in B cell and mast cell signal transduction and suggest that PKC-delta is important for the regulation of physiological B and mast cell functions as well as for their pathophysiology. Furthermore, dominant PKC-delta-independent effects of Rottlerin are presented, indicating restrictions of this inhibitor for use in signal transduction research. PMID- 11502861 TI - A novel proliferation-associated variant of CFR-1 defined by a human monoclonal antibody. AB - The germline coded human monoclonal IgM antibody 103/51 was isolated from a gastric carcinoma patient. This antibody binds to a 130-kd membrane molecule and has a mitotic effect on tumor cells in vitro. To characterize the target, we sequenced the protein and showed that the antibody binds to the cysteine-rich fibroblast growth factor receptor (CFR)-1, which is highly homologous to MG-160 and the E-selectin-ligand (ESL)-1. The epitope was determined by glycosidase digestion experiments to be an N-linked carbohydrate side chain. Immunohistochemistry was used to investigate the tissue distribution of CFR-1. Different healthy tissues were tested and only the collecting tubes of the kidney, the Golgi apparatus, and the glomerular and fascicular zones of the adrenal gland stained positive. However, on malignant tissue the receptor is overexpressed in nearly all tested stomach cancers (12 of 15) and other tested carcinomas (13 of 15). Most interestingly, the receptor is also present in Helicobacter pylori gastritis and gastric dysplasia, but absent on uninflamed stomach mucosa. This restricted tissue pattern indicates that antibody 103/51 reacts with a membrane-bound variant of CFR-1, which is mainly expressed on transformed cells and precursor lesions and is essential for proliferation processes. The possible activity of antibody 103/51 as an activating ligand in these proliferative changes of gastric epithelial mucosa is discussed. PMID- 11502862 TI - Inhibitors of phosphodiesterase isoforms III or IV suppress islet-cell nitric oxide production. AB - The general phosphodiesterase (PDE) inhibitor pentoxifylline (PTX), and the PDE type IV inhibitor rolipram (ROL), both increase intracellular cAMP levels and suppress inflammatory cytokine production by T cells and macrophages. We have previously shown that PTX and ROL protect from autoimmune diabetes in nonobese diabetic (NOD) mice. These drugs may mediate some of their anti-inflammatory effects by blocking nitric oxide (NO) production by macrophages. In this study, we investigated the effect of PDE inhibitors in blocking NO production by insulin secreting NIT-1 insulinoma cells and mouse islet cells in vitro and in vivo. Insulinoma cells and islet cells produced NO when stimulated with a combination of inflammatory cytokines and lipopolysaccharide (LPS). We found that both PTX and ROL markedly suppressed this induced NO production. Islet cells express PDEs III and IV and, accordingly, the PDE III inhibitor cilostamide (CIL) also suppressed NO production, and a combination of ROL and CIL had a synergistic effect. This suppression appeared to be mediated, at least in part, by elevating cAMP level and was mimicked by other cAMP-elevating agents, ie, membrane permeable cAMP analogs (dibutyryl cAMP and 8-bromo cAMP) and an adenylate cyclase stimulator (forskolin). PDE inhibitors suppressed the expression of inducible nitric oxide synthase (iNOS) mRNA. In vivo treatment with PTX or ROL prevented iNOS protein expression in the islets of NOD mice with cyclophosphamide accelerated disease. Our findings suggest that PDE inhibitors can protect islets against autoimmunity. PMID- 11502863 TI - Alpha-1-antitrypsin and a broad spectrum metalloprotease inhibitor, RS113456, have similar acute anti-inflammatory effects. AB - There is increasing evidence that antiproteases are able to affect the inflammatory response. To further examine this question, we administered human alpha-1-antitrypsin (alpha1AT) or a synthetic metalloprotease inhibitor (RS113456) to C57 mice followed by a single intratracheal dose of quartz, a dust that evokes a marked, lasting, polymorphonuclear leukocyte (PMN) infiltrate. At 2 hours after dust administration, both antiproteases completely suppressed silica induced PMN influx into the lung and macrophage inflammatory protein-2 (MIP 2)/monocyte chemotactic protein-1 (MCP-1) (neutrophil/macrophage chemoattractant) gene expression, partially suppressed nuclear transcription factor kappaB (NF kappaB) translocation, and increased inhibitor of NF-kappaB (IkappaB) levels. By 24 hours, PMN influx and connective tissue breakdown measured as lavage desmosine or hydroxyproline were still at, or close to, control levels after antiprotease treatment, and increases in NF-kappaB translocation and MIP-2/MCP-1 gene expression were variably suppressed. At both time points, neither agent prevented silica-induced increases in amount of whole lung MIP-2 or MCP-1 protein, but both did prevent increases in whole lung intercellular adhesion molecule-1 (ICAM-1) at 24 hours. Inactivating the alpha1AT by oxidation to the point that it no longer possessed antiproteolytic properties did not affect its ability to suppress inflammation. Both antiproteases also prevented the silica-induced acute inflammatory response in mice with knocked out genes for macrophage metalloelastase (MME -/-), mice that develop inflammation, but not connective tissue breakdown, and the pattern of alpha1AT breakdown fragments was identical in control and MME -/- animals. These findings suggest that, in this model of acute PMN mediated inflammation, a serine protease inhibitor and a metalloprotease inhibitor have similar anti-inflammatory properties, that inflammation is not mediated by proteolysis with generation of chemotactic matrix fragments, and that classic antiproteolysis (complexing of protease to antiprotease) probably does not play a role in suppression of inflammation. The antiproteolytic effects of these agents do not seem to be mediated by protection of endogenous alpha1AT. PMID- 11502864 TI - Overexpression of the granulocyte colony-stimulating factor gene impairs bone morphogenetic protein responsiveness in mice. AB - Granulocyte colony-stimulating factor (G-CSF) is the major hematopoietic growth factor regulating the production and differentiation of neutrophils. We previously demonstrated that permanent overexpression of G-CSF in transgenic mice produces a dramatic enlargement of the bone cavity and reduction of bone mass. This phenotype was shown to be associated with an increase of osteoclast-mediated bone resorption. As a way of determining the role of G-CSF in bone formation in vivo, an ectopic bone was induced subcutaneously into G-CSF transgenic mice by bone morphogenetic protein (BMP)-2, a potent initiator of bone and cartilage from undifferentiated mesenchymal cells. A BMP-2/atelocollagen pellet containing recombinant human BMP-2 was implanted into a dorsal subfascial pocket. At one week after implantation, proliferation of mesenchymal cells around the implant was significantly decreased in transgenic mice compared with control mice. At three weeks, an ectopic bone containing bone marrow was formed both in transgenic and control mice. However, the ectopic bones of the transgenic mice were smaller and less consistent than those of control mice, and the calcium contents were reduced to 56.2% of those of controls. The ectopic bone in the G-CSF mice showed poor development of both lamellar and trabecular bone. Semiquantitative reverse transcription-polymerase chain reaction analysis of the ectopic bone at 3 weeks disclosed no significant differences in the mRNA levels of type I collagen, osteopontin, and osteocalcin between G-CSF mice and control mice. Immunohistochemical study in G-CSF mice showed reduced staining of osteocalcin in the bone matrix surrounding the reduced number of osteoblasts. The half-life of BMP in the implants was prolonged to 7 to 9 days in the G-CSF mice, whereas it was 5 days in the control mice. Collectively, the permanent expression of G-CSF may retard the differentiation process of osteoblasts by impairing the initial induction of mesenchymal cells, resulting in reduction of bone mass, suggesting that G-CSF regulates the bone metabolism by modulating both osteoclast and osteoblast function. Furthermore, it is suggested that G-CSF is a potent modulator of the BMP-2 signal pathway in vivo. PMID- 11502865 TI - Macrophage-induced apoptosis limits endovascular trophoblast invasion in the uterine wall of preeclamptic women. AB - Impaired invasion of uteroplacental arteries by extravillous trophoblast cells is a key pathogenic mechanism of preeclampsia. We previously demonstrated that reduced trophoblast invasion into uteroplacental spiral arteries was associated with an excess of macrophages in and around these arteries. To explore the significance of these observations, we correlated the extent of extravillous trophoblast apoptosis in placental bed biopsy specimens with macrophage distribution and studied the effect of macrophages upon trophoblast apoptosis in vitro. Extravillous trophoblast hybrid cells were cocultured with activated macrophages exposed to exogenous tumor necrosis factor alpha (TNFalpha), anti tumor necrosis factor receptor I (TNF-RI), and tryptophan depletion, and the rates of trophoblast apoptosis were measured. Extravillous trophoblast hybrid cells showed increased rates of apoptosis following exposure to exogenous TNFalpha, with tryptophan depletion, and when cocultured with activated macrophages. The proapoptotic effects of macrophages in vitro were completely inhibited only by simultaneous addition of tryptophan and anti-TNF-RI. Our data indicate that macrophages, residing in excess in the placental bed of preeclamptic women, are able to limit extravillous trophoblast invasion of spiral arterial segments through apoptosis mediated by the combination of TNFalpha secretion and tryptophan depletion. The mechanisms by which macrophages are activated and recruited to the placental bed are presently unknown but are likely central to the pathogenesis of preeclampsia. PMID- 11502866 TI - Uterine natural killer cells in a three-dimensional tissue culture model to study trophoblast invasion. AB - The high numbers of CD56(+) cells with natural killer (NK) functions present in the uterine mucosa during the late secretory phase of the menstrual cycle and during early pregnancy have been considered to be implicated in implantation and in the regulation of trophoblast invasion. A three-dimensional organ culture model was used to study the interactions of these uterine NK cells with Jeg-3 and BeWo choriocarcinoma cells as a model of the invasive trophoblast. For this purpose, fragments of endometrial and decidual tissue were put in close contact with multicellular spheroids of choriocarcinoma cells in small silicon funnels. After the formation of stable contacts, the confrontation cultures were transferred to spinner flasks, cultivated for up to 6 days, and prepared for immunohistochemistry. During 2 days of cocultivation, the first cells started to move forward into the stromal component of the confrontation culture as demonstrated by staining of the choriocarcinoma cells using anti-human cytokeratin. Invasion advanced until, after a total of 6 days, some choriocarcinoma cells had already penetrated deeply into the host tissue. After a cultivation period of 1 week, both the endometrial and decidual tissue fragments still contained several CD56(+) uterine NK cells, and some of them expressed the proliferation-associated marker Ki-67 without any exogenous activation. A few CD56(+) cells were found directly at the invasion front, as well as between the choriocarcinoma cells. These cells also contained the cytolytic granule protein perforin indicating a migration of NK cells with cytolytic potential toward the potentially invasive cells. In conclusion, this human system closely resembles the in vivo conditions during trophoblast invasion and provides an appropriate in vitro model for studying dynamic processes involving various cell types during trophoblast invasion at the experimental level. Moreover, it enables us to study the effects of cytokines and growth factors that possibly regulate trophoblast invasion. PMID- 11502867 TI - Ki-Mcm6, a new monoclonal antibody specific to Mcm6: comparison of the distribution profile of Mcm6 and the Ki-67 antigen. PMID- 11502868 TI - An optimized protocol for first strand cDNA synthesis from laser capture microdissected tissue. PMID- 11502869 TI - Mass spectrometry of the proteome. PMID- 11502870 TI - Identification of the P2Y(12) receptor: a novel member of the P2Y family of receptors activated by extracellular nucleotides. PMID- 11502871 TI - Induction of L-arginine transport is inhibited by atrial natriuretic peptide: a peptide hormone as a novel regulator of inducible nitric-oxide synthase substrate availability. AB - BACKGROUND: The inducible nitric-oxide synthase (iNOS) synthesizes NO from L arginine. Availability of L-arginine is maintained by a lipopolysaccharide (LPS) induced induction of the CAT-2B amino acids transporter. Recently, we could show that the cardiovascular hormone atrial natriuretic peptide (ANP) inhibits the induction of iNOS in LPS-stimulated macrophages via its guanylate cyclase-coupled A-receptor. PURPOSE: To investigate whether ANP exerts an effect on LPS-induced L arginine uptake. METHODS: Murine bone marrow derived macrophages were activated with LPS (1 microg/ml, 20 h) in the presence or absence of ANP or C-type natriuretic peptide (CNP). L-Arginine transport was determined by measuring the uptake of L-[(3)H]arginine. L-[(3)H]Arginine influx was also determined in LPS activated cells in the presence of N(G)-monomethyl-L-arginine (L-NMMA), competitor amino acids, or ANP. Nitrite accumulation was determined in supernatants of LPS-activated cells cultured in the presence or absence of L ornithine. RESULTS: ANP dose dependently (10(-8)-10(-6)M) inhibited LPS-induced L [(3)H]arginine uptake when added simultaneously with LPS, whereas it showed no effect when added simultaneously with L-[(3)H]arginine. The effect was abrogated by the A-receptor antagonist HS-142-1 (10 microg/ml). CNP (10(-6) M) did not influence L-arginine transport. Competitor amino acids (10(-2) M) inhibited L [(3)H]arginine uptake. An excess of unlabeled L-arginine (10(-2) M) as well as its analog L-NMMA (10(-3) M) also reduced L-[(3)H]arginine influx. L-Arginine uptake was critical for production of NO because L-ornithine significantly decreased LPS-induced nitrite accumulation. CONCLUSION: This work demonstrates that ANP inhibits LPS-induced L-arginine uptake via its guanylate cyclase-coupled A-receptor. Besides its influence on the induction of iNOS, this effect may represent an important and unique mechanism by which ANP regulates NO production in macrophages. PMID- 11502872 TI - Regulation of the human CYP2B6 gene by the nuclear pregnane X receptor. AB - Cytochromes P450 (P450s) are involved in the oxidative metabolism of a plethora of structurally unrelated compounds, including therapeutic drugs. Two orphan members of the nuclear receptor superfamily, the pregnane X receptor (PXR; NR1I2) and constitutive androstane receptor (CAR; NR1I3) have been implicated in this phenomenon. In the present study, we examined the transcriptional regulation of the human CYP2B6 gene. In primary cultures of human hepatocytes, CYP2B6 was highly inducible by a number of compounds known to be human PXR ligands, including rifampicin and hyperforin. PXR was shown to be capable of activating the phenobarbital-responsive enhancer module (PBREM) region of the CYP2B6 gene, a 51-base-pair enhancer element that mediates induction of CYP2B6 expression by CAR. The two nuclear receptor-binding motifs within the PBREM effectively bound PXR as a heterodimer with the 9-cis retinoic acid receptor alpha (NR2B1). Taken together, these observations demonstrate that the CYP2B6 gene is directly regulated by PXR and further establish this receptor as a key regulator of drug metabolizing P450s. PMID- 11502873 TI - Molecular cloning of the platelet P2T(AC) ADP receptor: pharmacological comparison with another ADP receptor, the P2Y(1) receptor. AB - Platelet activation plays an essential role in thrombosis. ADP-induced platelet aggregation is mediated by two distinct G protein-coupled ADP receptors, Gq linked P2Y(1), and Gi-linked P2T(AC), which has not been cloned. The cDNA encoding a novel G protein-coupled receptor, termed HORK3, was isolated. The HORK3 gene and P2Y(1) gene were mapped to chromosome 3q21-q25. HORK3, when transfected in the rat glioma cell subline (C6-15), responded to 2-methylthio-ADP (2MeSADP) (EC(50) = 0.08 nM) and ADP (EC(50) = 42 nM) with inhibition of forskolin-stimulated cAMP accumulation. 2MeSADP (EC(50) = 1.3 nM) and ADP (EC(50) = 18 nM) also induced intracellular calcium mobilization in P2Y(1)-expressing cells. These results show that HORK3 is a Gi/o-coupled receptor and that its natural ligand is ADP. AR-C69931 MX and 2MeSAMP, P2T(AC) antagonists, selectively inhibited 2MeSADP-induced adenylyl cyclase inhibition in HORK3-expressing cells. On the other hand, A3P5PS, a P2Y(1) antagonist, blocked only 2MeSADP-induced calcium mobilization in P2Y(1)-expressing cells. HORK3 mRNA was detected in human platelets and the expression level of HORK3 was equivalent to that of P2Y(1). These observations indicate that HORK3 has the characteristics of the proposed P2T(AC) receptor. We have also determined that [(3)H]2MeSADP binds to cloned HORK3 and P2Y(1). Competition binding experiments revealed a similarity in the rank orders of potency of agonists and the selectivity of antagonists as obtained in the functional assay. These results support the view that P2Y(1) functions as a high-affinity ADP receptor and P2T(AC) as a low-affinity ADP receptor in platelets. PMID- 11502874 TI - Differential cell death induced by salsolinol with and without copper: possible role of reactive oxygen species. AB - Salsolinol (SAL), a novel dopaminergic catechol tetrahydroisoquinoline neurotoxin, has been speculated to contribute to the etiology of Parkinson's disease and neuropathology of chronic alcoholism. Our previous studies have demonstrated that SAL induces strand scission in oX174 supercoiled DNA and oxidative base modification in calf thymus DNA in the presence of cupric ion. We now report that treatment of rat pheochromocytoma (PC12) cells with SAL causes reduced viability, which was exacerbated by Cu(2+). The copper chelator bathocuproinedisulfonic acid ameliorated cytotoxicity induced by SAL and Cu(2+). N-Acetyl-L-cysteine and reduced glutathione protected against SAL- plus Cu(2+) mediated PC12 cell death. Cells exposed to SAL underwent apoptosis, as revealed by characteristic morphological and biochemical changes. SAL treatment resulted in increased levels of Bax with a concomitant decrease in expression of Bcl-x(L). Furthermore, SAL rapidly activated c-Jun N-terminal kinase, whereas the activity of extracellular signal-regulated protein kinase remained unchanged. Transfection with Bcl-x(L) or Bcl-2 led to protection against SAL-mediated PC12 cell death. Although SAL alone could cause apoptotic death in PC12 cells, cells treated with SAL together with Cu(2+) became necrotic. Cells exposed to both SAL and Cu(2+) exhibited higher levels of intracellular reactive oxygen species, malondialdehyde, and 8-oxo-7,8-dihydro-2'-deoxyguanosine than did those treated with SAL alone. These results suggest that copper accelerates redox cycling of SAL, leading to massive production of reactive oxygen species, which can divert the SAL-induced cell death to necrosis. PMID- 11502875 TI - Apoptosis induced by the homocamptothecin anticancer drug BN80915 in HL-60 cells. AB - The homocamptothecin (hCPT) derivative BN80915 containing a seven-membered lactone ring represents one of the most potent topoisomerase I inhibitors described. This anticancer agent, currently undergoing phase I clinical trials, has been shown to produce a greater number of DNA strand breaks than conventional camptothecins with a six-membered lactone ring. To shed light on the mechanism of action of hCPT at the cellular level, we compared the effects of BN80915 and the classic camptothecin SN-38, the active metabolite of irinotecan, on HL-60 human promyelocytic cancer cells. A variety of biochemical events, at both the mitochondrial and the nuclear levels, were characterized to determine how and to what extent the hCPT derivative can induce apoptotic cell death. The use of cytometry, Western blot analysis, confocal microscopy, and different colorimetric assays enabled us to demonstrate that BN80915 is a potent inducer of apoptosis in HL-60 cells. This induction of apoptosis is associated with cell cycle changes, a marked decrease of intracellular pH, activation of caspase-3 and -8, DNA fragmentation, and externalization of phosphatidylserine lipids but no significant changes of the mitochondrial membrane potential or the expression of Bcl-2. The interconnections between these different events are discussed. Collectively, the results indicate that the superior activity expressed at the topoisomerase I level leads to a more pronounced induction of apoptosis by BN80915 compared with SN-38. The study identifies and delineates signaling factors involved in BN80915-induced apoptosis in HL-60 cells. PMID- 11502876 TI - A Na(+)/Cl(-)-dependent transporter for catecholamines, identified as a norepinephrine transporter, is expressed in the brain of the teleost fish medaka (Oryzias latipes). AB - We report the isolation, functional characterization, and localization of a Na(+)/Cl(-)-dependent catecholamine transporter (meNET) present in the brain of the teleost fish medaka. This carrier is very similar to the human neuronal norepinephrine transporter (NET) and the human neuronal dopamine transporter (DAT), showing 70 and 64% amino acid identity, respectively. When expressed in COS-7 cells, this transporter mediates the high-affinity uptake of dopamine (K(M) = 290 nM) and norepinephrine (K(M) = 640 nM). Its pharmacological profile reveals more similarities with NET, including a high affinity for the tricyclic antidepressants desipramine (IC(50) = 0.92 nM) and nortriptyline (IC(50) = 16 nM). In situ hybridization on the medaka brain shows that meNET mRNA is present only in a subset of tyrosine hydroxylase-positive neurons found in the noradrenergic areas of the hindbrain, such as the locus ceruleus and area postrema. None of the dopaminergic areas anterior to the isthmus contains any labeled neurons. Neither reverse transcriptase-polymerase chain reaction with degenerate primers specific for gamma-aminobutyric acid transporter/NET nor autoradiographic experiments with [(125)I]3b-(4-iodophenyl)-tropane-2b-carboxylic acid methyl ester revealed an additional catecholamine transporter in the medaka brain. Uptake experiments with medaka brain synaptosomes show an endogenous transport with a pharmacological profile identical to that of the recombinant meNET. Thus, meNET is probably the predominant--if not the only--catecholamine transporter in the medaka fish brain. In view of the highly conserved primary structures and pharmacological properties of meNET, it is tempting to speculate that a specific dopamine transport developed later in vertebrate evolution and probably accompanied the tremendous enlargement of the meso-telencephalic dopaminergic pathways in amniotes. PMID- 11502877 TI - Inhibition of thymidylate synthase activity by antisense oligodeoxynucleotide and possible role in thymineless treatment. AB - Thymidylate synthase (TS) is an important target for chemotherapeutic treatment of cancer. However, efficacy of TS-targeted anticancer drugs is limited by the development of drug resistance as a result of TS gene amplification. In this work, a phosphorothioated antisense oligonucleotide (ODN), designated ATS-2, was used to suppress cellular synthesis of TS. ATS-2 at 0.2 microM concentration was mixed with lipofectin in a charge ratio of 1:1 and was used to treat the human embryonic kidney (HEK) cell line. A reduction of TS mRNA and protein was achieved. Furthermore, a dose-dependent reduction of cumulative viable cells of up to 98% was observed. Flow cytometer analysis of cell cycle progression indicates that ATS-2-treated cells were arrested and went into apoptosis at the S phase, possibly because of thymidine shortage, suggesting that ATS-2 is specifically effective for dividing cells. When used in combination with the anticancer drug FdUrd, ATS-2 exerted a additive inhibitory effect on cellular proliferation. To elucidate the possible role of cellular thymidine kinase (TdR kinase) in ATS-2 treatment, a second cell line, HeLa, was used. Both HEK and HeLa have similar rates of cell division and ODN uptake. In contrast to HEK, which was shown to have very low levels of TdR kinase activity in [(3)H]thymidine incorporation experiments, [(3)H]thymidine incorporation in HeLa was 15-fold greater than that of HEK. We found that HeLa cells were sensitive to FdUrd but were rather resistant to ATS-2. On the contrary, HEK cells were sensitive to ATS 2 but insensitive to FdUrd. Effects of ATS-2 and FdUrd are, therefore, complementary in thymineless treatment too. PMID- 11502878 TI - Glycosylation of the human prostacyclin receptor: role in ligand binding and signal transduction. AB - Prostacyclin, a potent vasodilator and inhibitor of platelet aggregation, acts through a cell-surface G protein-coupled receptor [prostacyclin (IP)]. The human (h) IP contains two consensus sites for N-linked glycosylation (N(7) and N(78)). However, the role of glycosylation is unknown. Mutant receptors (N(7)-Q(7),N(78) Q(78) and N(7),N(78)-Q(7),Q(78)) were generated by replacing N(7) and/or N(78) with Q's. Receptor glycosylation was similar in the wild-type and N(7)-Q(7) and was inhibited with tunicamycin. N(78)-Q(78) and N(7),N(78)-Q(7),Q(78) demonstrated little or no glycosylation. Membrane localization was reduced for each mutant concomitant with impaired glycosylation. Partial localization to the plasma membrane allowed direct examination of the effect of glycosylation on IP function. High-affinity binding to N(7)-Q(7) was similar (K(d) = 21.7 +/- 1.7 nM, n = 4) to that of the wild-type receptor (K(d) = 24.3 +/- 3.6 nM, n = 4), despite a reduced value for B(max) (0.35 +/- 0.03 fmol/mg of protein versus 3.34 +/- 0.52 fmol/mg of protein, n = 4). Binding to N(78)-Q(78) (B(max) = 0.27 +/- 0.03 fmol/mg of protein, n = 3; K(d) = 149.1 +/- 11.1, n = 3) and N(7),N(78) Q(7),Q(78) (no specific binding) was further impaired. Agonist-induced adenylyl cyclase activation was reduced in N(7)-Q(7) cells, whereas N(78)-Q(78) cells responded only to high concentrations of iloprost and N(7),N(78)-Q(7),Q(78) were unresponsive. Inositol phosphate generation was evident only with the wild-type. Only the wild-type and N(7)-Q(7) receptors underwent agonist-induced sequestration. Our findings demonstrate greater glycosylation at N(78) compared with N(7). The extent of N-linked glycosylation of hIP may be important for membrane localization, ligand binding, and signal transduction. PMID- 11502879 TI - Glutathione peroxidase-1 overexpression prevents ceramide production and partially inhibits apoptosis in doxorubicin-treated human breast carcinoma cells. AB - Reduced glutathione and N-acetylcysteine can inhibit both apoptosis and necrosis of several cell types, suggesting a critical role for reactive oxygen species (ROS) in cell death. However, how the cellular defense against oxidative stress is connected with other cell death mediators remains unclear. We selectively investigated the interaction of seleno-glutathione peroxidase-1 (GPx-1), the major enzyme responsible for peroxide detoxification in mammalian cells, with the cytotoxic response of T47D human breast cancer cells to doxorubicin, an anticancer drug known to promote production of ROS and apoptotic mediator ceramide. The sensitivity to doxorubicin-mediated cell death was compared in T47D/H3 containing low levels of endogenous GPx and T47D/GPx2 transfectant cells, which overexpress GPx-1. We show that T47D/GPx2 cells were significantly more resistant than T47D/H3 cells to doxorubicin (1 microM). The glutathione precursor, N-acetylcysteine also partially protected T47D/H3 cells from the lethal effect of doxorubicin, whereas L-buthionine-(S,R)-sulfoximine, an inhibitor of glutathione biosynthesis, sensitized both GPx-1--deficient and proficient cells. Interestingly, in addition to a decrease in ROS production, the activation of neutral sphingomyelinase, sphingomyelin hydrolysis, and ceramide generation in response to doxorubicin was impaired in T47D/GPx2 cells compared with control cells. In contrast, GPx overexpression did not protect breast cancer cells from cell death induced by exogenous cell-permeant ceramide. Moreover, the basal activity of neutral sphingomyelinase was considerably lower in T47D/GPx2. Taken together, these results indicate that GPx-1 can regulate doxorubicin induced cell death signaling at least in part by interfering with the activation of the sphingomyelin-ceramide pathway. PMID- 11502880 TI - Interaction of barbiturate analogs with the Torpedo californica nicotinic acetylcholine receptor ion channel. AB - Barbiturate-induced anesthesia is a complex mechanism that probably involves several ligand-gated ion channel superfamilies. One of these superfamilies includes the archetypical nicotinic acetylcholine receptor (nAChR), in which barbiturates act as noncompetitive antagonists. In this regard, we used the Torpedo californica nAChR and a series of barbiturate analogs to characterize the barbiturate binding site(s) on this superfamily member. [(14)C]Amobarbital binds to one high-affinity (K(d) = 3.7 microM) and several (approximately 11) low affinity (K(d) = 930 microM) sites on the resting and desensitized nAChRs, respectively. Characteristics of the barbiturate binding site on the resting nAChR include: (1) a tight structure-activity relationship. For example, the barbiturate isobarbital [5-ethyl-5'-(2-methylbutyl) barbituric acid] is >10-fold less potent than its formula isomer amobarbital [5-ethyl-5'-(3-methylbutyl) barbituric acid] in inhibiting [(14)C]amobarbital binding. (2) A binding locus within the pore of the nAChR ion channel. Each of the barbiturate analogs inhibited the binding of [(3)H]tetracaine or photoincorporation of 3 trifluoromethyl-3-(m-[(125)I]iodophenyl) diazirine in a mutually exclusive manner. (3) Stereoselective binding. The R(+)-enantiomers of isobarbital and pentobarbital are approximately 2-fold more potent in inhibiting 3 trifluoromethyl-3-(m-[(125)I]iodophenyl) diazirine photoincorporation than the S( )-enantiomers. Finally, molecular modeling suggests that within the channel, the pyrimidine ring of the barbiturate is located just above the highly conserved leucine ring (M2--9; e.g., delta Leu-265), whereas the 5' side chain projects downward, and depending upon its conformation, introduces steric hindrance to binding because of the restriction in the lumen of the channel introduced by the leucine side chains. PMID- 11502881 TI - Chinese herbal remedy wogonin inhibits monocyte chemotactic protein-1 gene expression in human endothelial cells. AB - Wogonin (Wog), an active component of Scutellaria baicalensis, has antioxidant and anti-inflammatory properties. Monocyte chemotactic protein-1 (MCP-1), a potent chemoattractant for monocytes, plays a crucial role in case of early inflammatory responses, including atherosclerosis. In this study, we investigated the effect of Wog on phorbol ester (PMA)-induced MCP-1 expression in human umbilical vein endothelial cells (ECs). The MCP-1 mRNA levels and MCP-1 release in Wog-treated ECs were measured. Wog inhibited PMA-induced MCP-1 mRNA levels and MCP-1 secretion in a dose-dependent manner. The inhibition of MCP-1 induction by Wog is a transcriptional event, as shown by Wog's significant reduction of both MCP-1 promoter and 4x 12-O-tetradecanoylphorbol-13-acetate response element luciferase reporter activities. By electrophoretic mobility assay, Wog significantly reduced the AP-1 binding activity induced by PMA. Furthermore, the PMA-induced extracellular signal-regulated kinase 1/2 and c-Jun amino-terminal kinase activities that contributed to AP-1 activity and MCP-1 gene induction were obviously attenuated after pretreating ECs with Wog. The decrease of MCP-1 secretion by Wog pretreatment led to a reduction of monocyte adhesion to ECs. Taken together, our results demonstrate that Wog inhibits MCP-1 induction in ECs; this inhibition is mediated by reducing AP-1 transcriptional activity via the attenuation of ERK1/2 and JNK signal transduction pathways. We conclude that Wog has the potential therapeutic development for use in anti-inflammatory and vascular disorders. PMID- 11502882 TI - cAmp modulates exocytotic kinetics and increases quantal size in chromaffin cells. AB - The role of cAMP/cAMP-dependent protein kinase (PKA) on the late phase of exocytosis has been studied by amperometry on Ba(2+)-stimulated single bovine chromaffin cells. Forskolin (FSK) increases the intracellular cAMP levels in a concentration-dependent manner. Forskolin (100 nM) does not increase the number of exocytotic events, although it significantly increases the net granule content of catecholamines (CA), which is accompanied by a slowing of the process of degranulation. These effects are reversible, occur within 15 to 60 s, and are not due to newly synthesized CA. Isoprenaline, pituitary adenylate cyclase-activating polypeptide-38 or dB-cAMP reproduce FSK effects as does cholera toxin. The inhibition of phosphodiesterases with 3-isobutyl-1-methylxanthine mimics and potentiates the effect of FSK and isoprenaline. Rolipram and okadaic acid also produce a drastic increase in net granule content of CA, whereas H-89 attenuates the FSK response. These data indicate that cyclic AMP/PKA might favor the granule aggregation before its fusion with cell membrane and slow the late step of the exocytotic process. PMID- 11502883 TI - Carrier-mediated delivery improves the efficacy of 9-(2 phosphonylmethoxyethyl)adenine against hepatitis B virus. AB - We recently synthesized a lipophilic prodrug of 9-(2-phosphonyl methoxyethyl)adenine (PMEA), designated PMEA-LO, and incorporated it into reconstituted lactosylated high-density lipoprotein (LacNeoHDL). In a rat model, LacNeoHDL-associated PMEA-LO was internalized by the asialoglycoprotein receptor on parenchymal liver cells and converted into its active diphosphorylated metabolite. To further evaluate the therapeutic potential of the carrier associated prodrug, we examined in this study the processing of (125)I-labeled PMEA-LO--loaded LacNeoHDL by HepG2 cells. Upon incubation with HepG2 cells, PMEA LO--loaded LacNeoHDL became rapidly cell-associated. The association was saturable and of high-affinity (k(d) = 3.8 +/- 0.4 nM). Asialofetuin, an established ligand for the asialoglycoprotein receptor, inhibited the association by >75%, which confirms the role of the asialoglycoprotein receptor. Association of the prodrug-loaded particles to HepG2 cells was coupled to degradation. Radiolabeled degradation products appeared in the culture medium with a lag phase of 2 h. Asialofetuin and chloroquine inhibited secretion of degradation products by 75 to 80%, indicating that PMEA-LO--loaded LacNeoHDL is internalized via the asialoglycoprotein receptor and lysosomally processed. The therapeutic potential of LacNeoHDL-associated PMEA-LO was studied by measuring its effects on hepatitis B virus (HBV) replication in Hep AD38 cells (HBV-transfected HepG2 cells). LacNeoHDL-associated PMEA-LO effectively inhibited HBV DNA synthesis. The EC(50) value of carrier-associated PMEA-LO was 35 times lower than that of free PMEA (3.4 +/- 0.4 and 120 +/- 18 ng of PMEA/ml, respectively). We conclude that the present results, combined with our earlier in vivo disposition data, underscore the therapeutic potential and utility of PMEA-LO--loaded LacNeoHDL for treatment of chronic hepatitis B. PMID- 11502884 TI - The green tea polyphenol (-)-epigallocatechin-3-gallate blocks nuclear factor kappa B activation by inhibiting I kappa B kinase activity in the intestinal epithelial cell line IEC-6. AB - The I kappa B kinase complex (IKK) mediates activation of the transcription factor nuclear factor-kappa B (NF-kappa B). We previously showed that green tea polyphenols inhibited endotoxin-mediated tumor necrosis factor-alpha (TNF alpha) production by blocking NF-kappa B activation. In this study, we evaluated whether green tea polyphenols inhibit NF-kappa B by blocking IKK activity. We assessed IKK activity by detecting changes in phosphorylation of an I kappa B alpha glutathione S-transferase (GST) fusion protein. IEC-6 cells pretreated with an extract of green tea polyphenols (GrTPs; 0--0.4 mg/ml) had diminished TNF alpha induced IKK and NF-kappa B activity. Of the various GrTPs, (-)-epigallocatechin-3 gallate (EGCG) was the most potent inhibitor. We next examined whether EGCG inhibited activated IKK. In cytosolic extracts of TNF alpha-stimulated cells, EGCG inhibited phosphorylation of I kappa B alpha-GST (IC(50) > 18 microM) consistent with inhibition of IKK activity. Using other polyphenols, we showed that the gallate group was essential for inhibition, and antioxidants were ineffective in blocking activated IKK. Importantly, EGCG decreased IKK activity in cytosolic extracts of NIK transiently transfected cells. This latter finding showed that our findings were not related to nonspecific kinase activity. In conclusion, EGCG is an effective inhibitor of IKK activity. This may explain, at least in part, some of the reported anti-inflammatory and anticancer effects of green tea. PMID- 11502885 TI - Identification of potent and selective neuropeptide Y Y(1) receptor agonists with orexigenic activity in vivo. AB - Neuropeptide Y (NPY) binds to a family of G-protein coupled receptors termed Y(1), Y(2), Y(3), Y(4), Y(5), and y(6). The use of various receptor subtype selective agonists and antagonists has facilitated identification of the receptor subtypes responsible for mediating many of the biological effects of NPY. For example, the potent orexigenic activity of NPY is believed to be mediated by both the Y(1) and Y(5) receptor subtypes. Several selective Y(5) receptor agonists that stimulate food intake in rodents are available, but no selective Y(1) receptor agonist has been reported. We have identified several NPY analogs that bind the NPY Y(1) receptor with high affinity and exhibit full agonist activity, measured as inhibition of forskolin-stimulated cAMP production in cells expressing the cloned NPY Y(1) receptor. [D-Arg(25)]-NPY, [D-His(26)]-NPY, Des AA(10--17)[Cys(7,21),Pro(34)]-NPY, Des-AA(11--18)[Cys(7,21),D-Lys(9)(Ac)]-NPY, Des-AA(11--18)[Cys(7,21),D-Lys(9)(Ac),Pro(34)]-NPY, Des-AA(11--18)[Cys(7,21),D Lys(9)(Ac),D-His(26)]-NPY and Des-AA(11--18)[Cys(7,21),D-Lys(9)(Ac),D-His(26), Pro(34)]-NPY bind the NPY Y(1) receptor with K(i) values of 0.9 +/- 0.2, 2.0 +/- 0.3, 0.2 +/- 0.05, 0.7 +/- 0.1, 0.2 +/- 0.01, 2.2 +/- 0.3, and 1.2 +/- 0.3 nM, respectively, and inhibit forskolin-stimulated cAMP production with EC(50) values of 0.2 +/- 0.02, 0.5 +/- 0.04, 0.3 +/- 0.03, 0.5 +/- 0.05, 0.4 +/- 0.16, 5.3 +/- 0.32, and 5.1 +/- 0.97 nM, respectively. These peptides are highly selective for the NPY Y(1) receptor relative to the NPY Y(2), Y(4), and Y(5) receptors. [D Arg(25)]-NPY, [D-His(26)]-NPY and Des-AA(11--18)[Cys(7,21), D-Lys(9)(Ac),D His(26),Pro(34)]-NPY stimulate food intake dose-responsively in Long-Evans rats for at least 4 h after intracerebroventricular administration. Although the involvement of Y(1) receptors in several physiological activities, such as vasoconstriction and anxiolysis, remains to be investigated, adequate tools are now available. PMID- 11502886 TI - 2-Aminoethoxydiphenyl borate directly inhibits store-operated calcium entry channels in human platelets. AB - In this study, we examined 2-aminoethoxydiphenyl borate (2APB) as an inhibitor of Ca(2+) influx in human platelets. 2APB was found to inhibit thrombin-mediated intracellular Ca(2+) mobilization rapidly in platelets incubated in the absence of extracellular Ca(2+). This result supports an intracellular action of 2APB on inositol 1,4,5-trisphosphate (IP(3))-receptor Ca(2+) channels. 2APB was without effect on the ability of thapsigargin to mobilize intracellular Ca(2+). This result suggests that the efflux of Ca(2+) from the endoplasmic reticulum mediated by thapsigargin is not via IP(3) Ca(2+) channels. However, 2APB was able to prevent the entry of Ca(2+) and Sr(2+) through thapsigargin-activated, store operated Ca(2+) channels (SOCC). This result supports a direct inhibitory effect of 2APB on SOCC. 2APB was also able to block the entry of Sr(2+), Ba(2+), and Mn(2+) entry into unstimulated platelets, which suggests that 2APB was inhibiting the Ca(2+) influx channels directly. The capacity of 2APB to prevent Ca(2+) influx and Sr(2+) influx was rapid because it occurred immediately upon addition to the platelets. The inhibition of Ca(2+) and Sr(2+) influx by 2APB was similar to that seen with the cell-impermeable nonselective Ca(2+)-channel blocker La(3+) or the Ca(2+) chelator EGTA. Diphenylboronic anhydride and 2,2 diphenyltetrahydrofuran, two compounds that are structurally similar to 2APB, also inhibited Ca(2+) influx. It was concluded that 2APB was a rapid and effective direct inhibitor of SOCC in human platelets; as such, it cannot be used to support the involvement of IP(3) receptors in the activation of SOCC. PMID- 11502887 TI - Antitumor activity of 2',3'-dideoxycytidine nucleotide analog against tumors up regulating DNA polymerase beta. AB - DNA polymerase beta (Pol beta), an error-prone DNA-synthesizing enzyme tightly down-regulated in healthy somatic cells, has been shown to be overexpressed in many human tumors. In this study, we show that treatment with the 2',3' dideoxycytidine (ddC) nucleoside analog inhibited in vitro and in vivo the proliferation of Pol beta-transfected B16 melanoma cells, which up-regulate Pol beta compared with control isogenic cells. The administration of ddC also increased specifically the survival of mice bearing Pol beta-overexpressing B16 melanoma. When the phosphorylated form of ddC was electrotransfered into Pol beta transfected melanoma, the cell growth inhibition was strengthened, strongly suggesting that the cytotoxic effect results from incorporation of the chain terminator into DNA. Using in vitro single- and double-stranded DNA synthesis assays, we demonstrated that excess Pol beta perturbs the replicative machinery, favors ddC-TP incorporation into DNA, and consequently promotes chain termination. Therefore, the use of chain terminator anticancer agents could be suitable for the treatment of tumors with a high level of Pol beta. PMID- 11502888 TI - Reversal of physiological stress-induced resistance to topoisomerase II inhibitors using an inducible phosphorylation site-deficient mutant of I kappa B alpha. AB - Physiological stress conditions associated with the tumor microenvironment play a role in resistance to anticancer therapy. In this study, treatment of EMT6 mouse mammary tumor cells with hypoxia or the chemical stress agents brefeldin A (BFA) or okadaic acid (OA) causes the development of resistance to the topoisomerase II inhibitor etoposide. The mechanism of physiological stress-induced drug resistance may involve the activation of stress-responsive proteins and transcription factors. Our previous work shows that treatment with BFA or OA causes activation of the nuclear transcription factor NF-kappa B. Pretreatment with the proteasome inhibitor carbobenzyoxyl-leucinyl-leucinyl-leucinal inhibits stress-induced NF-kappa B activation and reverses BFA-induced drug resistance. To test whether NF-kappa B specifically mediates stress-induced drug resistance, an inducible phosphorylation site-deficient mutant of I kappa B alpha (I kappa B alpha M, S32/36A) was introduced into EMT6 cells. In this study, we show that I kappa B alpha M expression inhibits stress-induced NF-kappa B activation and prevents BFA-, hypoxia-, and OA-induced resistance to etoposide. These results indicate that NF-kappa B activation mediates both chemical and physiological drug resistance to etoposide. Furthermore, they imply that coadministration of agents that inhibit NF-kappa B may enhance the efficacy of topoisomerase II inhibitors in clinical cancer chemotherapy. PMID- 11502889 TI - Agonist regulation of rat alpha 3 beta 4 nicotinic acetylcholine receptors stably expressed in human embryonic kidney 293 cells. AB - Effects of agonists on rat alpha 3 beta 4 nicotinic acetylcholine receptors expressed in KX alpha 3 beta 4R2 cells [human embryonic kidney 293-derived cells] were studied. The potencies of seven agonists varied over a 7000-fold range, with a rank order of epibatidine >> A85380 > cytisine approximately 1,1-dimethyl-4 phenyl-piperazinium iodide (DMPP) approximately nicotine > acetylcholine > carbachol. The efficacies of all of the agonists studied here were similar except for DMPP, which seemed to be a partial agonist compared with nicotine and acetylcholine. Nicotine and carbachol desensitized the receptors in a time- and concentration-dependent manner. The EC(50) values for nicotine and carbachol to desensitize the receptors during a 60-min exposure were 3 and 51 microM, respectively, indicating that these agonists are more potent at desensitizing the receptors than at activating them. The function of the receptors recovered from agonist-induced desensitization rapidly and almost completely. The half-time for recovery of function from desensitization after a 60-min treatment with nicotine increased with the concentration of nicotine used to desensitize the receptors. In contrast, no such concentration dependence for time to recovery of function was found when carbachol was used to desensitize the receptors. We propose that this difference may be due to the cell permeability of nicotine, allowing it to enter and be sequestered inside of cells and then slowly diffuse out to maintain receptor desensitization. After a 5-day exposure to 100 microM nicotine, the receptors were completely desensitized, but receptor function recovered to 83% of control values with a half-time of about 10.5 min. Although the number of nicotinic receptor binding sites measured with (+/-)-[(3)H]epibatidine was increased during the chronic treatment with nicotine, no increase in function was detected. PMID- 11502890 TI - Beta-adrenergic receptor subtype-specific signaling in cardiac myocytes from beta(1) and beta(2) adrenoceptor knockout mice. AB - The sympathetic nervous system modulates cardiac contractility and rate by activating beta-adrenergic receptors (beta AR) expressed on cardiac myocytes and specialized cells in the sinoatrial node and the conduction system. Recent clinical studies have suggested that beta-adrenergic receptors also play a role in cardiac remodeling that occurs in the pathogenesis of cardiomyopathy. Both beta(1) and beta(2) adrenergic receptors are expressed in human and murine hearts. We have examined the effect of beta AR activation on the spontaneous contraction rate of neonatal myocyte cultures from wild-type and beta receptor knockout (KO) mice (beta(1)AR-KO, beta(2)AR-KO and beta(1)beta(2)AR-KO mice). Stimulation of the beta(1)AR in beta(2)AR-KO myocytes produces the greatest increase in contraction rate through a signaling pathway that requires protein kinase A (PKA) activation. In contrast, stimulation of the beta(2)AR in beta(1)AR KO myocytes results in a biphasic effect on contraction rate with an initial increase in rate that does not require PKA, followed by a decrease in rate that involves coupling to a pertussis toxin sensitive G protein. A small isoproterenol induced decrease in contraction rate observed in beta(1)beta(2)AR-KO myocytes can be attributed to the beta(3)AR. These studies show that all three beta AR subtypes are expressed in neonatal cardiac myocytes, and the beta(1)AR and beta(2)AR couple to distinct signaling pathways. PMID- 11502891 TI - The Effects of isoflurane on acetylcholine receptor channels: 3. Effects of conservative polar-to-nonpolar mutations within the channel pore. AB - We performed macroscopic and single-channel current measurements on wild-type (WT) and two mutant muscle-type nicotinic acetylcholine (ACh) receptor channels transiently expressed in HEK-293 cells. The mutants contained polar-to-nonpolar substitutions at the 10' (alpha(2)S10'A beta T10'A gamma delta) and 6' positions (alpha(2)S6'A beta gamma delta S6'A) in the M2 pore region of the channel. We studied the behavior of these channels in the absence and presence of the volatile general anesthetic isoflurane. Both mutations changed the gating behavior of the channel. A comparison of the alpha(2)S10'A beta T10'A gamma delta mutant to WT receptors revealed faster desensitization kinetics, increased sensitivity to ACh, a higher efficacy for activation by the partial nicotinic agonist decamethonium, and a greater number of openings per burst. A comparison of the alpha(2)S6'A beta gamma delta S6'A mutant to WT receptors also revealed increased sensitivity to ACh and an increased burst duration at the single channel level with ACh as agonist. The alpha(2)S10'A beta T10'A gamma delta mutation increased the sensitivity of the ACh receptor to isoflurane, whereas the alpha(2)S6'A beta gamma delta S6'A mutation did not. These changes were probably not caused by the differential effects of the mutation on channel gating and desensitization. The increased sensitivity of the alpha(2)S10'A beta T10'A gamma delta receptor to isoflurane is state-dependent; the mutation changes the affinity of the closed state but not that of the open state of the channel. PMID- 11502892 TI - Estrogen receptor alpha mediates the proliferative but not the cytotoxic dose dependent effects of two major phytoestrogens on human breast cancer cells. AB - Phytoestrogens are a chemically diverse group of compounds made by plants that can have estrogenic effects in animals. Both tumorigenic and antitumorigenic effects have been reported. Although estrogens stimulate the growth of many breast tumors, there is a negative correlation between the incidence of breast cancer and the phytoestrogen-rich diet of certain Asian populations. To begin to resolve this paradox, we have analyzed the estrogenic properties of genistein and quercetin, two flavonoid phytoestrogens particularly abundant in soybeans. Trans activation experiments with a transfected reporter gene for nuclear estrogen receptors (ER) show strong activation of the endogenous ER alpha by both phytoestrogens in two MCF7 human breast cancer cell lines. This is supported by the observation that the two phytoestrogens induce the down-regulation of ER alpha mRNA and protein levels. Using chimeric proteins consisting of the hormone binding domains of ER alpha and ER beta fused to the Gal4 DNA binding domain, we have established that genistein and quercetin are full estrogenic agonists of both ER isoforms. Ligand binding experiments with purified ER alpha and ER beta confirm that the two phytoestrogens are ER ligands. At concentrations that are sufficient to obtain substantial transcriptional activity, they stimulate the proliferation of two ER alpha-dependent breast cancer cell lines. At high concentrations, such as those reached with a soy-rich diet, genistein and quercetin are strong cytotoxic agents that even kill ER-independent HeLa cells. Thus, the mode of action of phytoestrogens and the balance between being risk or chemopreventive factors for breast cancer may depend on the dietary load. PMID- 11502893 TI - Ca(v)3.2 channel is a molecular substrate for inhibition of T-type calcium currents in rat sensory neurons by nitrous oxide. AB - Although nitrous oxide (N(2)O; laughing gas) remains widely used as an anesthetic and analgesic in clinical practice, its cellular mechanisms of action remain inadequately understood. In this report, we examined the effects of N(2)O on voltage-gated Ca(2+) channels in acutely dissociated small sensory neurons of adult rat. At subanesthetic concentrations, N(2)O blocks low-voltage-activated, T type Ca(2+) currents (T currents), but not high-voltage-activated (HVA) currents. This blockade of T currents was concentration dependent, with an IC(50) value of 45 +/- 13%, maximal block of 38 +/- 12%, and Hill coefficient of 2.6 +/- 1.0. No desensitization of the response or change in current kinetics was observed during N(2)O application. The magnitude of T current blockade by N(2)O does not seem to reflect any use- or voltage-dependent properties. In addition, T current blockade was not altered when intracellular GTP was replaced with guanosine 5'-(gamma thio)triphosphate or guanosine 5'-0-(2-thiodiphosphate) suggesting a lack of involvement of G-proteins in the inhibition. N(2)O selectively blocked currents arising from the Ca(v)3.2 but not Ca(v)3.1 recombinant channels stably expressed in human embryonic kidney (HEK) cells in a concentration-dependent manner with an apparent affinity and potency similar to native dorsal root ganglion currents. Analogously, the block of Ca(v)3.2 T currents exhibited little voltage- or use dependence. These data indicate that N(2)O selectively blocks T-type but not HVA Ca(2+) currents in small sensory neurons and Ca(v)3.2 currents in HEK cells at subanesthetic concentrations. Blockade of T currents may contribute to the anesthetic and/or analgesic effects of N(2)O. PMID- 11502894 TI - Regulation of the rat glutathione S-transferase A2 gene by glucocorticoids: involvement of both the glucocorticoid and pregnane X receptors. AB - Glucocorticoids regulate the rat glutathione S-transferase A2 (GSTA2) gene in a biphasic manner in cultured hepatocytes that repress gene expression at low concentration (10--100 nM) but induce gene expression at high concentration (>1 microM). High concentrations of the glucocorticoid receptor (GR) antagonist RU38486 (5--10 microM) also induced the expression of GSTA2. These effects were reproduced in HepG2 cells transfected with a luciferase reporter containing 1.6 kilobase pairs of 5'-flanking sequence of GSTA2 and expression plasmids for either GR, pregnane X receptor (PXR) or a combination of both. Dexamethasone t butylacetate (1 microM t-Bu-DEX) repressed gene expression between 60 to 75% when only GR was expressed. When PXR was expressed, both basal and t-Bu-DEX-dependent gene expression was increased over 2-fold, respectively. Biphasic regulation of gene expression was observed over a broad range of t-Bu-DEX concentrations when expression plasmids for both receptors were cotransfected. Other steroids of the pregnane class induced GSTA2 expression as expected for a PXR-dependent process. Because no canonical responsive element for the PXR-RXR alpha heterodimer was observed in the 5'-flanking region of the construct, deletion analysis was used to identify a pregnane responsive region between base pairs -700 and -683; this 20-bp region contains the antioxidant response element (ARE). When the ARE sequence was mutated, basal, t-butylhydroquinone- and 17 alpha hydroxypregnenolone-inducible expression were all lost. These results suggest that PXR interacts with factors binding to the ARE to elicit the pregnane inductive response for GSTA2. PMID- 11502896 TI - A cue to queue for CoQ? PMID- 11502895 TI - A phenylalanine residue at segment D3-S6 in Nav1.4 voltage-gated Na(+) channels is critical for pyrethroid action. AB - Mammalian voltage-gated Na(+) channels were less sensitive to pyrethroids than their insect counterparts by 2 to 3 orders of magnitude. Deltamethrin at 10 microM elicited weak gating changes in rat skeletal muscle alpha-subunit Na(+) channels (Nav1.4) after > 30 min of perfusion. About 10% of the peak current was maintained during the 8-ms, +50-mV pulse and, upon repolarization to -140 mV, the amplitude of the slow tail current corresponded to less than 3% of total Na(+) channels modified by deltamethrin. A background mutation, Nav1.4-I687M (within D2:S4-S5 cytoplasmic linker), enhanced the deltamethrin-induced maintained current by approximately 2.5-fold, whereas Nav1.4-I687T, a homologous superkdr mutation, reduced it by approximately 2-fold. Repetitive pulses at 2 Hz further augmented the effects of deltamethrin on Nav1.4-I687M mutant channels so that approximately 75% of peak currents were maintained. A second mutation, Nav1.4 I687M/F1278I at the middle of D3-S6, rendered the channel greatly resistant to deltamethrin. This double mutant channel remained sensitive to batrachotoxin (BTX), even though nearby residues S1276 and L1280 were critical for BTX action. We hypothesize that the deltamethrin receptor and the BTX receptor are situated at the middle but opposite surface of the D3-S6 alpha-helical structure. Another mutant, Nav1.4-I687M/N784K, exhibited a partial deltamethrin-resistant phenotype but was completely resistant to BTX. Consistent with the BTX-resistant phenotype of N784K and the known adjacent kdr mutation at position L785F, deltamethrin and BTX were probably situated next to each other upon binding at D2-S6. Evidently, distinct residues from multiple S6 segments were critical for deltamethrin and BTX actions. PMID- 11502897 TI - Chromosome 22q and the mind-brain interface: bedside to bench to bedside. PMID- 11502898 TI - Changing cortical excitability with low-frequency magnetic stimulation. PMID- 11502899 TI - Nisus steps forth. PMID- 11502901 TI - Training clinical researchers in neurology: we must do better. AB - The pace of scientific discoveries, the increasing complexity of managing patients, and the runaway cost of neurological services have created an urgent need for a wide range of clinical research in neurology. Despite increasing recognition of this need and recent increases in funding for training clinical investigators, neurologists conducting cellular and molecular investigations are more likely to join faculties, maintain research careers, and attain academic advancement. Because academic departments of neurology are successful in producing and nurturing basic science researchers, why aren't they just as triumphant in spawning clinical investigators? This crisis in the preparation of clinical investigators has been brought about by many factors: competing time demands for clinical service, lack of methodologically rigorous training in the disciplines necessary to conduct clinical research, and lack of mentorship. Neurology residents contemplating a clinical research career may observe junior faculty who lack career guidance, are ill-prepared as independent investigators, and must juggle patient demands while trying to write a research grant or conduct a study. Already burdened by medical school debts, is it any wonder that our neurology graduates don't leap to a career with a future that seems so insecure? Academic departments of neurology must develop full-scale clinical research training programs if they are to meet the pressing need for clinical research. As a starting point, they must free themselves from their dependence on providing clinical services to generate income. Following the model which has produced successful basic researchers, much greater effort must be given to establishing rigorous methodological training in collaboration with other departments, creating senior role models, and protecting time for clinical investigators to conduct research. Unless we create incentives to careers in clinical research, we will never answer the growing number of clinical research questions we face today. PMID- 11502902 TI - Symmetric intracerebral calcifications. PMID- 11502903 TI - A randomized, placebo-controlled trial of coenzyme Q10 and remacemide in Huntington's disease. AB - OBJECTIVES: To determine whether chronic treatment with coenzyme Q10 or remacemide hydrochloride slows the functional decline of early Huntington's disease (HD). METHODS: The authors conducted a multicenter, parallel group, double-blind, 2 x 2 factorial, randomized clinical trial. Research participants with early HD (n = 347) were randomized to receive coenzyme Q10 300 mg twice daily, remacemide hydrochloride 200 mg three times daily, both, or neither treatment, and were evaluated every 4 to 5 months for a total of 30 months on assigned treatment. The prespecified primary measure of efficacy was the change in total functional capacity (TFC) between baseline and 30 months. Safety measures included the frequency of clinical adverse events. RESULTS: Neither intervention significantly altered the decline in TFC. Patients treated with coenzyme Q10 showed a trend toward slowing in TFC decline (13%) over 30 months (2.40- versus 2.74-point decline, p = 0.15), as well as beneficial trends in some secondary measures. There was increased frequency of nausea, vomiting, and dizziness with remacemide and increased frequency of stomach upset with coenzyme Q10. CONCLUSIONS: Neither remacemide nor coenzyme Q10, at the dosages studied, produced significant slowing in functional decline in early HD. PMID- 11502904 TI - Limb immobilization for the treatment of focal occupational dystonia. AB - BACKGROUND: Occupational focal upper-limb dystonia is characterized by involuntary muscle contractions that selectively interfere with the execution of specific motor tasks such as writing or playing a musical instrument. Occupational dystonias have a severe social impact, especially in certain professions. The available medical treatments offer little benefit. METHODS: In eight patients with idiopathic occupational focal dystonia of the upper limb, the dystonic forearm and hand were immobilized with a plastic splint for mean (+/-SD) 4.5 +/- 0.75 weeks. Before splinting (base line) and at various intervals afterwards (4, 12, and 24 weeks), the authors assessed the severity of dystonia and the patients' motor performance objectively (Arm Dystonia Disability Scale and Tubiana and Chamagne Score) and subjectively (Self-Rating Score). RESULTS: Assessment 4 weeks after splint removal, when patients had regained normal voluntary movements, showed that the severity of dystonia and the patients' performance of the impaired motor task had improved; the benefit persisted unchanged at later follow-up visits (Arm Dystonia Disability Scale: base line 20.6 +/- 30.2%; after 4 weeks 83.9 +/- 23.8%, p = 0.007; after 12 weeks 83.9 +/- 23.8%, p = 0.007; after 24 weeks 79.7 +/- 29.5%, p = 0.015. Tubiana and Chamagne Score: base line 28.6 +/- 22.7%; after 4 weeks 80.0 +/- 23.1%, p = 0.015; after 12 weeks 80.0 +/- 23.1%, p = 0.015; after 24 weeks 74.3 +/- 32.1%, p = 0.031. Self-Rating Score: base line 20.6 +/- 19.3%; after 4 weeks 63.7 +/- 25.2%, p = 0.015; after 12 weeks 66.9 +/- 28.1%, p = 0.015; after 24 weeks 70.6 +/- 31.8%, p = 0.015). At the 24-week visit the improvement disappeared in one patient, was moderate in three, and marked in four. CONCLUSIONS: Limb immobilization can be a simple, effective, safe, and inexpensive treatment for focal occupational upper limb dystonia. PMID- 11502905 TI - Catecholamines in patients with 22q11.2 deletion syndrome and the low-activity COMT polymorphism. AB - OBJECTIVE: To investigate catecholamine phenotypes and the effects of a tyrosine hydroxylase inhibitor in individuals with the 22q11.2 deletion syndrome and low activity catechol-O-methyltransferase (COMT). BACKGROUND: Many persons with the 22q11.2 deletion syndrome suffer severe disability from a characteristic ultrarapid-cycling bipolar disorder and associated "affective storms." One etiologic hypothesis for this condition is that deletion of the COMT gene from one chromosome 22 results in increased catecholamine neurotransmission, particularly if the undeleted chromosome 22 encodes a variant of COMT with low activity. METHODS: In a preliminary study, plasma, urine, and CSF catecholamines and catecholamine metabolites were measured in four teenage patients with a neuropsychiatric condition associated with 22q11.2 deletion and the low-activity COMT polymorphism on the nondeleted chromosome. In these four patients, and an additional institutionalized adult with the condition, an uncontrolled, open label trial of metyrosine was administered in an attempt to lower catecholamine production and to alleviate symptoms. RESULTS: Mild elevations of baseline CSF homovanillic acid (HVA) were found in three of four patients and a moderate reduction in CSF HVA after metyrosine treatment in the patient with the highest pretreatment concentration. The course of the five patients during the clinical trial is described. CONCLUSIONS: In patients with the 22q11.2 deletion syndrome and low-activity COMT, controlled studies of pharmacologic agents that decrease catecholamine production, block presynaptic catecholamine storage, or enhance S adenosylmethionine, the cosubstrate of COMT, are warranted. PMID- 11502906 TI - LIS1 missense mutations cause milder lissencephaly phenotypes including a child with normal IQ. AB - BACKGROUND: Classical lissencephaly is a disorder of neuroblast migration with most patients having mutations of either the LIS1 or DCX genes. Most patients with lissencephaly secondary to LIS1 mutations have a severe malformation consisting of generalized agyria and pachygyria. However, increasing experience suggests that the phenotypic spectrum is wider than previously thought. METHODS: The authors describe the clinical and imaging features and mutation data of the five known patients with missense mutations of the LIS1 gene and emphasize one patient with normal intelligence. RESULTS: Patients with a missense mutation of the LIS1 gene have a wider and milder spectrum of cortical malformations and clinical sequelae compared with patients with other mutation types. CONCLUSION: Milder and more variable phenotypes seen in patients with missense mutations of LIS1 are likely a consequence of suboptimal function of the mutant LIS1 protein, rather than complete loss of function of this protein. The authors suggest that the few patients found thus far with missense mutations of LIS1 results from an underascertainment of patients with more subtle malformations and that abnormalities of the LIS1 gene may account for a greater spectrum of neurologic problems in childhood than has previously been appreciated. PMID- 11502907 TI - Oculomotor abnormalities in attention deficit hyperactivity disorder: a preliminary study. AB - BACKGROUND: Prevailing hypotheses suggest that attention deficit hyperactivity disorder (ADHD) is secondary to dysfunction of motor intentional systems mediated by prefrontal circuitry. Oculomotor paradigms provide a mechanism for examining and localizing dysfunction at the interface between movement and cognition. OBJECTIVE: Three different saccade tasks (reflexive or prosaccades, antisaccades, and memory-guided saccades) were used to examine functions necessary for the planning and the execution of eye movements, including motor response preparation, response inhibition, and working memory. METHODS: The study included 19 children with ADHD, divided into two groups: a group of 8 children on methylphenidate at the time of testing and a group of 11 children not taking any psychoactive medication. Results from the two groups were compared with those from 25 age- and gender-matched normal control children. RESULTS: Both groups of children with ADHD made significantly more directional errors than did controls on the antisaccade task and significantly more anticipatory errors than did controls on the memory-guided saccade task, findings that are consistent with deficits in response inhibition. There were no significant differences in prosaccade latency, although unmedicated children with ADHD showed significantly greater variability in latency on the prosaccade task than did controls. On the memory-guided saccade task there were no significant differences in saccade accuracy; however, unmedicated children with ADHD showed longer saccade latency than did either controls or medicated children with ADHD. CONCLUSIONS: Oculomotor findings suggest that deficits in prefrontal functions, in particular response inhibition, contribute to behavioral abnormalities observed in ADHD. Findings also suggest that the administration of methylphenidate is associated with improvements in the consistency of motor response. Although there were no observed improvements in response inhibition with methylphenidate, conclusions await a design in which subjects complete testing both on and off medication. PMID- 11502908 TI - Sudden unexplained death in children with epilepsy. AB - BACKGROUND: Sudden unexplained death is a significant cause of mortality in people with epilepsy. Risk factors that have been identified include male sex, poor compliance with medications, and antiepileptic drug (AED) polypharmacy. However, these may not apply to the pediatric population in which the causes of epilepsy differ from the adult population. Therefore, risk factors for sudden unexplained death in epilepsy (SUDEP) in children must be evaluated independently from those in the adult population. METHODS: Cases of SUDEP in children less than 18 years of age occurring over a 10-year period in the province of Ontario, Canada, were identified. Records were reviewed for demographic and clinical features and neuropathology findings. RESULTS: Twenty-seven cases of SUDEP in children were identified. Sixty-three percent were male. Age at death ranged from 8 months to 15 years. Fourteen children had symptomatic epilepsy (52%), five had cryptogenic epilepsy (18%), and eight had idiopathic epilepsy (30%). Twelve children were treated with one AED (46%), 10 were on two AED (38%), and three were on three AED (12%). At the time of death, seven children had one serum AED concentration below the therapeutic range (35%) and 12 children had AED levels within the therapeutic range (60%). CONCLUSIONS: This case series represents the largest series of sudden unexplained death in children with epilepsy. At least two previously described risk factors for SUDEP in adults, low serum AED levels at time of death and AED polytherapy, do not appear to be significant in children. PMID- 11502909 TI - Increased driving accident frequency in Danish patients with epilepsy. AB - OBJECTIVE: To determine driving accident frequency in a cohort of patients with epilepsy. METHODS: A 10-year historical cohort register study of 159 subjects with epilepsy and 559 controls individually matched for age, gender, place of residence, and exposure period was carried out. All had nonprofessional driver's licenses without restrictions. Persons with recorded diagnoses of other neurologic diseases, diabetes, psychoses, seizures, abuse, or poisoning of any kind were not included. The outcome measure was treatment at the casualty department after an accident as a car driver. RESULTS: Ten patients with epilepsy and five controls had been treated at the casualty department, the rate per 1,000 person-years with exposure being seven times higher (CI 2.18 to 26.13) in those with epilepsy than in the control cohort. CONCLUSIONS: Drivers with epilepsy are more likely than healthy controls to be treated at a casualty department after having a motor vehicle accident. PMID- 11502910 TI - Early hormonal changes during valproate or carbamazepine treatment: a 3-month study. AB - BACKGROUND: Long-term treatment with valproate (VPA) or carbamazepine (CBZ) may induce reproductive endocrine disorders in patients with epilepsy. METHODS: Serum concentrations of reproductive hormones were studied in 17 women and 22 men with recently diagnosed epilepsy before they started either VPA or CBZ medication, and 1 and 3 months later. RESULTS: No weight gain or clinical signs of hormonal disorders were observed during the follow-up. The mean serum levels of testosterone, luteinizing hormone, follicle-stimulating hormone, and sex hormone binding globulin (SHBG) increased, and dehydroepiandrosterone sulfate (DHEAS) decreased, in women starting VPA. Serum testosterone levels increased in half of the women on VPA. Serum concentrations of progesterone and dehydroepiandrosterone increased, and gonadotropins decreased, in men on VPA during the follow-up. Serum SHBG levels increased and DHEAS decreased during the first months of CBZ treatment in both sexes. In addition, the free-androgen index decreased in men after starting CBZ. CONCLUSIONS: Hormonal changes occur after only 1 month's use of VPA or CBZ. VPA-treatment seems to be associated with increased serum androgen levels, but the profile of hormonal changes appears to be different in women than in men. The use of CBZ, in turn, was associated with increased SHBG concentrations and thus with diminished sex steroid function in both sexes. The women with increased serum testosterone levels in the early phase of VPA medication may be at increased risk for VPA-related endocrine disorders later during treatment. PMID- 11502911 TI - Decreased bone mass and increased bone turnover with valproate therapy in adults with epilepsy. AB - BACKGROUND: Bone loss and hypovitaminosis D are reported in patients taking antiepileptic drugs, but little is known about changes in bone and calcium metabolism from valproic acid (VPA). OBJECTIVE: To assess the relationship of VPA to bone mass and calcium metabolism in 40 adults with epilepsy on long-term VPA monotherapy, 40 age- and sex-matched epileptic patients taking phenytoin (PHT), and 40 healthy control subjects. Bone mineral density (BMD) of the second metacarpal was determined as T- and Z-scores. RESULTS: BMD reduction from control values was 14% (12% in men, 16% in women) with VPA and 13% (12% in men, 15% in women) with PHT. Among patients on VPA, nine (23%) had T-scores below -2.5 SD, suggesting osteoporosis; 15 (37%) had T-scores between -1 and -2.5 SD, suggesting osteopenia. Serum concentrations of calcium were significantly higher with VPA than in PHT or control groups. Serum concentrations of bone Gla protein (a bone formation marker) and pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP; a bone resorption marker) associated with either drug significantly exceeded control values. Z-scores for BMD in the VPA group correlated negatively with calcium and ICTP. High ICTP correlated positively with ionized calcium, implying that increased bone resorption caused the latter. CONCLUSION: Long-term VPA monotherapy can increase bone resorption, leading to decreased BMD. PMID- 11502912 TI - Decreased corticospinal excitability after subthreshold 1 Hz rTMS over lateral premotor cortex. AB - OBJECTIVE: To study whether trains of subthreshold 1 Hz repetitive transcranial magnetic stimulation (rTMS) over premotor, prefrontal, or parietal cortex can produce changes in excitability of motor cortex that outlast the application of the train. BACKGROUND: Prolonged 1 Hz rTMS over the motor cortex can suppress the amplitude of motor-evoked potentials (MEP) for several minutes after the end of the train. Because TMS can produce effects not only at the site of stimulation but also at distant sites to which it projects, the authors asked whether prolonged stimulation of sites distant but connected to motor cortex can also lead to lasting changes in MEP. METHODS: Eight subjects received 1500 magnetic stimuli given at 1 Hz over the left lateral frontal cortex, the left lateral premotor cortex, the hand area of the left motor cortex, and the left anterior parietal cortex on four separate days. Stimulus intensity was set at 90% active motor threshold. Corticospinal excitability was probed by measuring the amplitude of MEP evoked in the right first dorsal interosseous muscle by single suprathreshold stimuli over the left motor hand area before, during, and after the conditioning trains. RESULTS: rTMS over the left premotor cortex suppressed the amplitude of MEP in the right first dorsal interosseous muscle. The effect was maximized (approximately 50% suppression) after 900 pulses and outlasted the full train of 1500 stimuli for at least 15 minutes. Conditioning rTMS over the other sites did not modify the size of MEP. A control experiment showed that left premotor cortex conditioning had no effect on MEP evoked in the left first dorsal interosseous muscle. CONCLUSIONS: Subthreshold 1 Hz rTMS of the left premotor cortex induces a short-lasting inhibition of corticospinal excitability in the hand area of the ipsilateral motor cortex. This may provide a model for studying the functional interaction between premotor and motor cortex in healthy subjects and patients with movement disorders. PMID- 11502913 TI - Frequency of bowel movements and the future risk of Parkinson's disease. AB - BACKGROUND: Constipation is frequent in PD, although its onset in relation to clinical PD has not been well described. Demonstration that constipation can precede clinical PD could provide important clues to understanding disease progression and etiology. The purpose of this report is to examine the association between the frequency of bowel movements and the future risk of PD. METHODS: Information on the frequency of bowel movements was collected from 1971 to 1974 in 6790 men aged 51 to 75 years without PD in the Honolulu Heart Program. Follow-up for incident PD occurred over a 24-year period. RESULTS: Ninety-six men developed PD an average of 12 years into follow-up. Age-adjusted incidence declined consistently from 18.9/10,000 person-years in men with <1 bowel movement/day to 3.8/10,000 person-years in those with >2/day (p = 0.005). After adjustment for age, pack-years of cigarette smoking, coffee consumption, laxative use, jogging, and the intake of fruits, vegetables, and grains, men with <1 bowel movement/day had a 2.7-fold excess risk of PD versus men with 1/day (95% CI: 1.3, 5.5; p = 0.007). The risk of PD in men with <1 bowel movement/day increased to a 4.1-fold excess when compared with men with 2/day (95% CI: 1.7, 9.6; p = 0.001) and to a 4.5-fold excess versus men with >2/day (95% CI: 1.2, 16.9; p = 0.025). CONCLUSIONS: Findings indicate that infrequent bowel movements are associated with an elevated risk of future PD. Further study is needed to determine whether constipation is part of early PD processes or is a marker of susceptibility or environmental factors that may cause PD. PMID- 11502914 TI - Time trends in the incidence of parkinsonism in Olmsted County, Minnesota. AB - OBJECTIVE: To investigate time trends in the incidence of parkinsonism and PD over a 15-year period (1976 to 1990). METHODS: The authors used the medical records-linkage system of the Rochester Epidemiology Project to identify incidence cases of parkinsonism in Olmsted County, MN, over three 5-year periods, 1976 to 1980, 1981 to 1985, and 1986 to 1990. PD and other types of parkinsonism were classified using defined criteria. Population denominators were derived from census data and were corrected by removing prevalent cases of parkinsonism. RESULTS: Over the 15 years of the study, 364 cases of parkinsonism were identified; 154 (42%) of them had PD. The incidence of parkinsonism remained stable over the three 5-year periods for the age classes 0 to 39, 40 to 59, and 60 to 69 years. For the age class 70 to 99 years, there was some increase over time mainly owing to an increased incidence of drug-induced parkinsonism. The incidence of PD remained stable over the three 5-year periods for all age classes. Results were similar when considering men and women separately. No birth cohort effect was present for parkinsonism. Comparison with three previous studies in the same population did not reveal any major long-term secular trends in the incidence of parkinsonism. CONCLUSIONS: The findings for PD over 15 years and comparison of the findings with historical data for parkinsonism over half a century suggest that no major environmental risk factors for PD (e.g., environmental toxins, drugs, diet constituents, or infectious agents) were introduced or removed from this population during these periods. PMID- 11502915 TI - Randomized placebo-controlled trial of donepezil in patients with progressive supranuclear palsy. AB - OBJECTIVE: There is no effective treatment for progressive supranuclear palsy (PSP). Because results of immunochemical and pharmacologic studies suggest that the cholinergic system may play a role in the cognitive and motor features of PSP, the authors investigated the effects of donepezil (10 mg/day), an acetylcholinesterase inhibitor, in 21 patients with PSP (mean age +/- SD; 65.7 +/ 4.7 years) by a randomized, double-blind, placebo-controlled crossover trial. METHODS: Donepezil and placebo were administered for 6 weeks each with a 1-month washout period. Patients were evaluated before and at the end of each treatment phase. Outcome measures evaluated neuropsychiatric, global cognitive, frontal, memory, motor, and activities of daily living (ADL) status. RESULTS: Two patients withdrew during the washout phase because of unrelated medical problems. Donepezil-induced systemic side effects were transient and generally mild. Because of worsening of motor function, three patients received 5 mg/day of donepezil. All patients achieved blood and CSF therapeutic levels of donepezil. While the patients were taking donepezil, their Double Memory Test scores improved, whereas their ADL/mobility scores significantly worsened. CONCLUSION: The findings suggest that acetylcholinesterase inhibitors such as donepezil have at best selective, modest effects on cognition in patients with PSP. In light of its deleterious effects on ADL/mobility, donepezil is not recommended for this patient population. PMID- 11502916 TI - Predictors of good outcome after intravenous tPA for acute ischemic stroke. AB - BACKGROUND: Thrombolytic therapy for acute ischemic stroke with IV alteplase is increasingly well established in North America but not elsewhere. Baseline factors that altered the response to alteplase were not identified by the National Institute of Neurological Disorders and Stroke tPA Stroke Study Group. METHODS: The authors gathered information from centers in the United States, Canada, and Germany on 1,205 patients with acute ischemic stroke treated with IV alteplase. The purpose was to identify independent factors that were predictive of good outcome using multivariable logistic regression modelling. The modified Rankin Scale score was dichotomized into good outcome (mRS 0 to 1) and poor outcome (mRS >1) as the primary outcome measure. RESULTS: In relative order of decreasing magnitude, milder baseline stroke severity, no history of diabetes mellitus, normal CT scan, normal pretreatment blood glucose level, and normal pretreatment blood pressure were independent predictors of good outcome among patients treated with IV alteplase for acute ischemic stroke. Confounding was observed among history of diabetes mellitus, CT scan appearance, baseline serum glucose level, and blood pressure, suggesting important relationships among these variables. CONCLUSIONS: Several factors were independently predictive of good outcome among patients with acute ischemic stroke treated with alteplase. These results require further confirmation before clinical implementation. PMID- 11502917 TI - A 1-year, placebo-controlled preservation of function survival study of donepezil in AD patients. AB - OBJECTIVE: To examine the effects of donepezil compared with placebo on the preservation of function in patients with AD over a 1-year period. METHODS: This was a prospective, 54-week, double-blind, placebo-controlled, survival to endpoint study. Patients were required to have at entry: a diagnosis of probable AD (National Institute of Neurological and Communicative Disorders and Stroke criteria); Mini-Mental State Examination score of 12 to 20; Clinical Dementia Rating of 1 or 2; modified Hachinski ischemia score < or =4; and capability of performing 8 of 10 instrumental activities of daily living and 5 of 6 basic activities of daily living. Patients (n = 431) were randomized to placebo or donepezil (5 mg/day for 28 days, 10 mg/day thereafter). Outcome measures were the AD Functional Assessment and Change Scale, the Mini-Mental State Examination, and Clinical Dementia Rating scale. At each visit, investigators determined whether predefined criteria for clinically evident decline in functional status had been met. Patients who met the endpoint criteria were discontinued per protocol. RESULTS: Donepezil extended the median time to clinically evident functional decline by 5 months versus placebo. The probability of patients treated with donepezil remaining in the study with no clinically evident functional loss was 51% at 48 weeks, compared with 35% for placebo. The Kaplan-Meier survival curves for the two treatment groups were different (p = 0.002, log-rank test). CONCLUSIONS: Patients with AD continue to show detectable disease progression over time, but treatment with donepezil for 1 year was associated with a 38% reduction in the risk of functional decline compared with placebo. PMID- 11502918 TI - A 1-year, randomized, placebo-controlled study of donepezil in patients with mild to moderate AD. AB - OBJECTIVE: To evaluate the long-term clinical efficacy and safety of donepezil versus placebo over 1 year in patients with mild to moderate AD. METHODS: Patients (n = 286; mean age, 72.5 years) with possible or probable AD from five Northern European countries were randomized to receive either donepezil (n = 142; 5 mg/day for 28 days, followed by 10 mg/day) or placebo (n = 144) for 1 year. RESULTS: The study was completed by 66.9% of the donepezil- and 67.4% of the placebo-treated patients. The benefit of donepezil over placebo was demonstrated by the Gottfries-Brane-Steen (a global assessment for rating dementia symptoms) total score at weeks 24, 36, and 52 (p < 0.05) and at the study end point (week 52, last observation carried forward; p = 0.054). Advantages of donepezil over placebo were also observed in cognition and activities of daily living (ADL) assessed by the Mini-Mental State Examination at weeks 24, 36, and 52, and the end point (p < 0.02) and by the Progressive Deterioration Scale at week 52 and the end point (p < 0.05). Adverse events (AE) were recorded for 81.7% of donepezil- and 75.7% of placebo-treated patients, with 7% of donepezil- and 6.3% of placebo-treated patients discontinuing because of AE. Treatment response to donepezil was not predicted by APOE genotype or sex in this population. CONCLUSION: As the first 1-year, multinational, double-blinded, placebo controlled study of a cholinesterase inhibitor in AD, these data support donepezil as a well tolerated and effective long-term treatment for patients with AD, with benefits over placebo on global assessment, cognition, and ADL. PMID- 11502919 TI - The prevalence of neuropsychiatric syndromes in systemic lupus erythematosus. AB - OBJECTIVE: To describe the prevalence of neuropsychiatric (NP) syndromes in a Finnish population of patients with systemic lupus erythematosus (SLE) and to classify them according to the recently developed American College of Rheumatology (ACR) nomenclature and case definitions for NPSLE. METHODS: Cross sectional, population-based study covering an area with 440,000 people. A total of 58 patients with a definite diagnosis of SLE and aged 16 to 65 years were found in the computerized database of the area hospitals. Of these, 46 (79%) agreed to participate. The diagnosis of various NP syndromes was based on clinical impression (H.A.) following history, examination, review of medical records, and neuropsychologic testing. RESULTS: At least one NP syndrome was identified in 42 patients (91%). The most frequent manifestation was cognitive dysfunction (n = 37; 81%), followed by headache (n = 25; 54%) and mood disorder (n = 20; 43%). When mild NP syndromes (mild cognitive deficit, headache, mild depression, anxiety, electroneuromyography-negative polyneuropathy) were excluded, the prevalence of NPSLE dropped to 46%. CONCLUSIONS: According to the ACR nomenclature, there is a high prevalence of NP manifestations in a population based sample of patients with SLE. Most NP syndromes were classified as minor; if they were excluded, the 46% prevalence of NPSLE would be slightly less than estimated in previous studies. PMID- 11502920 TI - Current management of ALS: comparison of the ALS CARE Database and the AAN Practice Parameter. The American Academy of Neurology. AB - BACKGROUND: The American Academy of Neurology (AAN) ALS Practice Parameter was published in April 1999. The ALS CARE Database has been collecting data on the management of patients with ALS in North America since 1996. OBJECTIVE: To compare the management of patients with ALS in North America as recorded in the ALS CARE Database with the recommendations of the AAN ALS Practice Parameter. METHODS: Data were analyzed from 2018 patients at enrollment and from 373 of these patients who died between enrollment and May 1999. RESULTS: Eighty-two percent of the enrolled patients reported that they had been given enough information about ALS. Only 54% of patients with drooling were receiving medication for this problem. Only 41% of those who reported being depressed most of the time were receiving antidepressant medications. Only 28% of those with dyspnea and only 9.2% of those with a forced vital capacity <40% predicted were receiving noninvasive positive pressure ventilator support. Only 30% of those with moderate to severe dysphagia had a gastrostomy tube. Half of the patients who died did so at home, but only 47% of them received residential hospice services. Although 89% of patients who died were recorded as having done so peacefully, 17% were reported to have had breathing difficulties (i.e., respiratory distress), 8% anxiety, 3.3% pain, and 2.5% choking. Advance directives were in place for 90% of the patients who died, and in 97% of cases these directives were followed. CONCLUSIONS: These findings indicate that in the 3-year period prior to the publication of the AAN Practice Parameter, many but not all patients received the care that is recommended in that parameter; there were deficiencies, particularly in the key areas of gastrostomy and noninvasive positive pressure ventilation. PMID- 11502921 TI - Lamotrigine reduces painful diabetic neuropathy: a randomized, controlled study. AB - OBJECTIVE: To study the efficacy of lamotrigine in relieving the pain associated with diabetic neuropathy. METHODS: The authors randomly assigned 59 patients to receive either lamotrigine (titrated from 25 to 400 mg/day) or placebo over a 6 week period. Primary outcome measure was self-recording of pain intensity twice daily with a 0 to 10 numerical pain scale (NPS). Secondary efficacy measures included daily consumption of rescue analgesics, the McGill Pain Questionnaire (MPQ), the Beck Depression Inventory (BDI), the Pain Disability Index (PDI), and global assessment of efficacy and tolerability. RESULTS: Twenty-four of 29 patients (83%) receiving lamotrigine and 22 of 30 (73%) patients receiving placebo completed the study. Daily NPS in the lamotrigine-treated group was reduced from 6.4 +/- 0.1 to 4.2 +/- 0.1 and in the control group from 6.5 +/- 0.1 to 5.3 +/- 0.1 (p < 0.001 for lamotrigine doses of 200, 300, and 400 mg). The results of the MPQ, PDI, and BDI remained unchanged in both groups. The global assessment of efficacy favored lamotrigine treatment over placebo, and the adverse events profile was similar in both groups. CONCLUSIONS: Lamotrigine is effective and safe in relieving the pain associated with diabetic neuropathy. PMID- 11502922 TI - Poor Beard!! Charcot's internationalization of neurasthenia, the "American disease". AB - OBJECTIVE: To analyze the role of the seminal 19th-century neurologist, Jean Martin Charcot, in the internationalization of neurasthenia, previously known as "the American disease." BACKGROUND: The New York neurologist, George Beard, first described neurasthenia in 1869 and considered it a disorder related to the particular stress of modern civilization, most typified in the United States. METHODS: Charcot's personal files on neurasthenia from the Bibliotheque Charcot, Salpetriere Hospital, Paris, were studied and his teaching lessons and lectures were examined. RESULTS: Charcot presented numerous cases of neurasthenia, always crediting Beard with the original name and description. Calling Beard's 1880 work a "remarkable monograph," Charcot emphasized that patients with neurasthenia also comprised the bulk of his own private practice. Focusing on the signs of sexual impotency, fatigue, and a tight band of pressure around the head, Charcot categorically distinguished patients with neurasthenica from patients with hysteria. Like Beard, Charcot concluded that the origin of neurasthenia was psychological stress and felt the European society also fostered the environment to precipitate the disease. Charcot adamantly opposed extrapolations that called for early childhood educational reforms to reduce current classroom stress. Charcot sympathized more with the treating physician than the patient, calling neurasthenics insufferable (insupportables). On the front sheet of his neurasthenia file, he wrote in large script, "Poor Beard!!" CONCLUSION: By emphasizing the prevalence of neurasthenia and extending Beard's observations, Charcot internationally legitimized the new diagnosis. Adding neurasthenia to the other neurologic descriptions from the United States by Hammond, Mitchell, and Dana, Charcot helped to foster the recognition of the American Neurologic School. PMID- 11502923 TI - Coenzyme Q10 reverses pathological phenotype and reduces apoptosis in familial CoQ10 deficiency. AB - Two brothers with myopathic coenzyme Q10 (CoQ10) deficiency responded dramatically to CoQ10 supplementation. Muscle biopsies before therapy showed ragged-red fibers, lipid storage, and complex I + III and II + III deficiency. Approximately 30% of myofibers had multiple features of apoptosis. After 8 months of treatment, excessive lipid storage resolved, CoQ10 level normalized, mitochondrial enzymes increased, and proportion of fibers with TUNEL-positive nuclei decreased to 10%. The authors conclude that muscle CoQ10 deficiency can be corrected by supplementation of CoQ10, which appears to stimulate mitochondrial proliferation and to prevent apoptosis. PMID- 11502924 TI - Agnosia for familiar faces and odors in a patient with right temporal lobe dysfunction. AB - The authors studied a 53-year-old man with progressive prosopagnosia and inability to recognize his favorite foods by smell. He could not identify pictures of familiar faces, but he could match unfamiliar faces and distinguish them from familiar ones. He could not identify familiar odors, but he could detect them and could perceive them as pleasant or familiar. Neuroimaging revealed temporal lobe changes, especially on the right. Right temporal lesions may produce face and odor agnosia by preventing perceptual familiarity units from accessing semantic associations. PMID- 11502925 TI - Preservation of golf skills in a case of severe left lobar frontotemporal degeneration. AB - The authors report the longitudinal study of a 53-year-old man with severe lobar atrophy confined to the left frontal and temporal lobes, including the left hippocampus, but sparing other cortical regions. He experienced profound cognitive deterioration, sparing only visuospatial memory. Despite these deficits, he could play golf at a high level of competence, following rules and etiquette as well as monitoring the ongoing game. The patient's golf performance may have been supported by residual visuospatial declarative memory and complex flexible implicit memory programs. PMID- 11502926 TI - Persistent prolongation of simple reaction time in sports concussion. AB - A baseline computerized cognitive assessment was completed by 483 military cadets before their initial school year. Fourteen cadets concussed during physical education boxing were retested <1 hour after injury and again on return to full activity 4 days later. Compared with baseline testing, postinjury performance on simple reaction time and continuous performance tests was significantly slowed, even after cadets experienced resolution of physical symptoms and were cleared to resume full activity. These findings may be relevant to current concussion management guidelines. PMID- 11502927 TI - Dilated cervical epidural veins and extra arachnoid fluid collection in orthostatic headaches. AB - Cervical MRI of eight patients with severe orthostatic headache showed a convex shaped, prominent, dilated anterior, internal vertebral venous plexus with a flow void signal in all patients and in seven also extra arachnoid fluid collection. Follow-up MRI showed resolution of the abnormalities in all patients. The authors hypothesize that these findings reflect disturbed compensatory vascular mechanisms by which autoregulation of the CSF pressure tries to compensate for the reduced CSF volume. PMID- 11502928 TI - Delayed post-irradiation bulbar palsy in nasopharyngeal carcinoma. AB - In a hospital series of 70 patients on follow-up after radiotherapy for nasopharyngeal carcinoma, 14 patients (20%) developed delayed post-irradiation bulbar palsy 1 to 18 years after radiotherapy (mean 5.5 years). Functional disability was moderate to severe. Three patients had aspiration pneumonia with one mortality. Post-irradiation bulbar palsy was a common complication and probably resulted from direct neuronal damage. PMID- 11502929 TI - False lateralization by subdural electrodes in two patients with temporal lobe epilepsy. AB - The authors present two patients with medically refractory partial seizures who had invasive recordings with stereotactic depth EEG (SEEG) and subdural electrodes (SDE) as part of their presurgical workup. SDE recordings were falsely lateralizing in both of these patients with pathologically proven mesial temporal sclerosis. In temporal lobe epilepsy, SEEG electrodes should be considered when presurgical studies are discordant. PMID- 11502930 TI - Ketotic hyperglycemia and epilepsia partialis continua. AB - Epilepsia partialis continua (EPC) may occur during nonketotic hyperglycemia but has not been described with diabetic ketoacidosis. The authors report a patient with EPC associated with ketotic hyperglycemia. Brain MRI showed two areas of abnormal signal intensity in the left precentral gyrus and in the right cerebellar hemisphere. Hyperglycemia may reduce seizure threshold because of the increase in gamma-aminobutyric acid metabolism and may trigger epileptic discharges. PMID- 11502931 TI - Population study of benign rolandic epilepsy: is treatment needed? AB - Forty-three of 79 children (54%) with benign rolandic epilepsy from a regional population were treated with antiepileptic drugs (AED); 36 (46%) were not. Physician advice was a major determinant of treatment choice. AED significantly reduced generalized seizures (p = 0.001) but did not reduce partial seizures. After 4 to 14 years and >900 seizures, all patients were in remission without medication or injury. Physicians may confidently offer a no-AED treatment strategy. PMID- 11502932 TI - Narcolepsy associated with other central nervous system disorders. AB - The authors identified patients with the coexistence of narcolepsy and another CNS disorder seen between 1975 and 1998 at their institution. Eighteen patients were identified, nine with narcolepsy commencing within 1 year before or after the other disorder. Seven patients (39%) had hypothalamic-pituitary syndromes. When they occur together, narcolepsy and other CNS disorders frequently emerge at about the same time, suggesting a causative relationship. Hypothalamic-pituitary pathology was the most common association. PMID- 11502933 TI - CSF antiretroviral drug penetrance and the treatment of HIV-associated psychomotor slowing. AB - The authors evaluated whether highly active antiretroviral therapy (HAART) with multiple CSF-penetrating drugs results in greater improvement in HIV-associated psychomotor slowing than HAART with a single CSF-penetrating drug. Both groups had improvement in CD4 count, plasma viral load, as well as two tests of psychomotor speed. Comparing the two groups, there were no differences in the mean change for CD4 count, viral load, or any of the neuropsychological tests. Multiple and single CSF-penetrating HAART may be equivalent for treating HIV associated psychomotor slowing. PMID- 11502934 TI - Visual loss in cysticercosis: analysis of 23 patients. AB - Of 23 cases with cysticercal visual loss, 13 had optic neuropathy (due largely to papilledema), 4 sustained chiasmal damage, and 6 had retrochiasmal lesions. Papilledema usually was associated with hydrocephalus, chiasmal involvement was caused by inflammation and compression from adjacent cysts, and retrochiasmal damage was produced by large parenchymal cysts or vasculitic cerebral infarction. PMID- 11502935 TI - A prospective natural history study of inclusion body myositis: implications for clinical trials. AB - Eleven patients with untreated inclusion body myositis (IBM) were prospectively studied during a 6-month period that included muscle strength, lean body mass, and muscle mass measurements. There was an overall quantifiable mean decline in percent of predicted normal muscle strength of 4% from baseline in a 6-month period, but one third of patients showed no change or slight improvements in strength. Short-term treatment trials in IBM will require large numbers of patients to detect slowing, arrest, or even slight improvement in muscle strength. PMID- 11502936 TI - Delirium from nicotine withdrawal in neuro-ICU patients. AB - Five cases of presumed nicotine withdrawal delirium among brain-injured patients treated in a neurologic intensive care unit are presented. Each patient had a history of heavy tobacco use and experienced dramatic and sustained clinical improvement within hours of transdermal nicotine replacement. These preliminary observations suggest that nicotine withdrawal may be an under-recognized cause of delirium in patients with acute brain injury. PMID- 11502937 TI - Prevalence and correlates of parkinsonism in patients with primary depression. AB - The authors examined the prevalence, clinical correlates, and longitudinal changes of parkinsonism in 94 patients with primary depression and 20 healthy control subjects. Parkinsonism was present in 20% of patients with primary depression. This syndrome was significantly associated with older age, more severe depression, and more severe cognitive impairment. In a subgroup of depressed patients, parkinsonism was reversible upon recovery from the mood disorder. PMID- 11502938 TI - The professor or the resident? A misidentification of two faces. PMID- 11502939 TI - Bilateral pallidal deep brain stimulation for cervical and truncal dystonia. PMID- 11502940 TI - Systemic lupus erythematosus with anti-Hu antibodies and polyradiculoneuropathy. PMID- 11502941 TI - Creatine monohydrate does not increase strength in patients with hereditary neuropathy. PMID- 11502942 TI - Lack of association between ubiquitin carboxy-terminal hydrolase L1 gene polymorphism and PD. PMID- 11502944 TI - Weight change associated with valproate and lamotrigine monotherapy in patients with epilepsy. PMID- 11502945 TI - The FAB: a frontal assessment battery at bedside. PMID- 11502946 TI - Failure of long-term pallidal stimulation corrected by subthalamic stimulation in PD. PMID- 11502947 TI - Late-onset SCA2: 33 CAG repeats are sufficient to cause disease. PMID- 11502948 TI - Improving the delivery of care to the seriously mentally ill. PMID- 11502949 TI - Resolving disparities in antidepressant treatment and quality-of-life outcomes between uninsured and insured primary care patients with depression. AB - BACKGROUND: Efforts to improve primary care depression treatment should penetrate to vulnerable uninsured populations. OBJECTIVE: To assess a primary care intervention's impact on treatment and quality-of-life outcomes in uninsured and insured depressed patients during the acute treatment phase. RESEARCH DESIGN: Twelve community primary care practices were randomized to 'enhanced' (intervention) and usual care conditions. Physicians, nurses and administrative staff in enhanced care practices received training to improve detection and management of depression. SUBJECTS: In 1996 to 1997, 383 nonelderly depressed patients who were either uninsured or covered by private insurance/Medicaid were enrolled; 343 (89.6%) completed six-month follow-up. MEASURES: Adequate pharmacotherapy (>or=3 months of antidepressants at therapeutic doses); adequate psychotherapy (>or=8 counseling visits); improvement in mental-health-related quality-of-life (MHQOL), assessed by Mental Component Summary scale for SF-36. RESULTS: Multivariate results showed that 54.6% of uninsured enhanced care (UEC) patients received adequate pharmacotherapy, compared with 14.3% of uninsured usual care (UUC) patients (P = 0.0005); however, receipt of adequate psychotherapy was comparable between these two groups (18.2% UEC, 11.9% UUC; P = 0.42). Intervention effects on insured patients' treatment were modest to minimal. Among usual care patients, the insured had 5.4 points greater improvement in MHQOL at 6 months than the uninsured (12.4 points insured, 7.0 points uninsured; P = 0.02); however, among patients receiving the intervention, the insured and uninsured had comparable MHQOL improvement (12.3 points insured, 11.6 points uninsured; P = 0.76). CONCLUSIONS: The intervention improved antidepressant treatment rates in uninsured patients and helped resolve quality of-life outcome disparities observed between insured and uninsured patients receiving usual care. PMID- 11502950 TI - Use of pharmacy data to assess quality of pharmacotherapy for schizophrenia in a national health care system: individual and facility predictors. AB - OBJECTIVES: This paper presents a profile of the use of antipsychotic medications in the treatment of schizophrenia in a national health system. METHODS: Prescription drug records written for antipsychotic medications between June 1999 and September 1999 were collected for patients diagnosed with schizophrenia in the Department of Veteran Affairs (VA). Indicators were constructed describing whether patients received multiple antipsychotic medications and whether the total weekly dose was outside of the range specified in the treatment recommendations developed by the schizophrenia Patient Outcomes Research Team (PORT). Generalized estimation equations were used to identify patient and facility characteristics that are associated with adherence to PORT recommendations. RESULTS: Of the 34,925 patients in the final sample, 2,383 (6.8%) received prescriptions for more than one antipsychotic (polypharmacy). A higher number of patients (4,554 or 13.0%) were dosed above the PORT recommendations on an antipsychotic medication and even more (8,148 or 23.3%) were dosed below the recommended PORT dosage. Older patients, minorities, and those with comorbid depression or substance abuse were generally less likely to receive multiple antipsychotics or be dosed above PORT recommendations. Neither academic emphasis (the percentage of the mental health budget spent on research and education) nor fiscal stress was significantly associated with adherence to recommendations. CONCLUSIONS: In the nation's largest mental health system, a relatively small number of patients were prescribed multiple antipsychotic medications, but more than a third were dosed outside of the PORT recommended range. PMID- 11502951 TI - Can quality improvement programs for depression in primary care address patient preferences for treatment? AB - BACKGROUND: Depression is common in primary care, but rates of adequate care are low. Little is known about the role of patient treatment preferences in encouraging entry into care. OBJECTIVES: To examine whether a primary care based depression quality improvement (QI) intervention designed to accommodate patient and provider treatment choice increases the likelihood that patients enter depression treatment and receive preferred treatment. METHODS: In 46 primary care clinics, patients with current depressive symptoms and either lifetime or current depressive disorder were identified through screening. Treatment preferences, patient characteristics, and use of depression treatments were assessed at baseline and 6 months by patient self-report. Matched clinics were randomized to usual care (UC) or 1 of 2 QI interventions. Data were analyzed using logistic regression models. RESULTS: For patients not in care at baseline, the QI interventions increased rates of entry into depression treatment compared with usual care (adjusted percentage: 50.0% +/- 5.3 and 33.0% +/- 4.9 for interventions vs. 15.9% +/- 3.6 for usual care; F = 12.973, P <0.0001). Patients in intervention clinics were more likely to get treatments they preferred compared with those in usual care (adjusted percentage: 54.2% +/- 3.3 and 50.7% +/- 3.1 for interventions vs. 40.5% +/- 3.1 for usual care; F = 6.034, P <0.003); however, in all clinics less than half of patients preferring counseling reported receiving it. CONCLUSIONS: QI interventions that support patient choice can improve the likelihood of patients receiving preferred treatments. Patient treatment preference appears to be related to likelihood of entering depression treatment, and patients preferring counseling may require additional interventions to enhance entry into treatment. PMID- 11502952 TI - Medical surveillance after breast cancer diagnosis. AB - OBJECTIVES: To characterize the tests ordered for surveillance of breast cancer recurrence in the 4 years after breast cancer diagnosis by surgeons, medical oncologists, and radiation oncologists. RESEARCH DESIGN: 303 stage I or II breast cancer patients age 55-years or older and diagnosed at 1 of 5 Boston hospitals. Patient interviews and medical record abstracts provided the data to characterize patient demographics, the breast cancer stage and its primary therapy, and the surveillance procedures ordered. RESULTS: 279 of the 303 women had some surveillance testing. Among those who received some surveillance, a mean of 22.0 tests were ordered, most by their medical oncologists (mean = 14.4), followed by their surgeons (mean = 9.7) and their radiation oncologists (mean = 5.7). The most common test was a mammogram (mean = 3.9). Women ages 75 to 90 years old were at higher risk for failure to complete four consecutive years of surveillance and for receipt of less than guideline surveillance. Younger women, women treated at a breast cancer center with a unified patient chart, and women who worked full or part time were at lower risk for failure to complete 4 years of surveillance. CONCLUSION: Most women in this cohort received some surveillance after completing primary therapy for breast cancer. Although no woman's surveillance corresponded exactly to existing guidelines, the oldest women were least likely to receive guideline surveillance. Surveillance after breast cancer therefore joins the list of aspects of breast cancer care-breast cancer screening, diagnosis, prognostic evaluation, and primary therapy-for which older women receive less than definitive care. PMID- 11502953 TI - The treatment of neck and low back pain: who seeks care? who goes where? AB - BACKGROUND: Neck and low back pain are leading causes of morbidity and health care utilization. However, little is known about the characteristics that differentiate those who seek from those who do not seek health care for their pain. OBJECTIVES: The objectives of this study were to: 1) describe health care utilization for neck and back pain; 2) determine the characteristics of individuals seeking health care for neck and back pain; and 3) identify the characteristics of patients who consult medical doctors, chiropractors, or both. DESIGN: Population-based cross-sectional mailed survey. SUBJECTS: Subjects were randomly selected adults from the Saskatchewan Health Insurance and Registration File. MEASURES: Demographic, socio-economic, general health, comorbidity, health related-quality-of-life, pain severity and health care utilization data were collected. The main outcome was whether subjects with prevalent neck or low back pain visited a health care provider in the previous month. RESULTS: Twenty-five percent of individuals with neck or low back pain visited a health care provider. Seeking health care was associated with disabling neck or back pain, digestive disorders, worse bodily pain and worse physical-role-functioning. Compared with medical patients, fewer chiropractic patients lived in rural areas or reported arthritis, but they reported better social and physical functioning. More patients consulting both providers reported disabling neck or back pain. CONCLUSIONS: Individuals seeking care for neck or back pain have worse health status than those who do not seek care. Patients consulting chiropractors alone report fewer comorbidities and are less limited in their activities than those consulting medical doctors. PMID- 11502954 TI - Screening for high utilizing somatizing patients using a prediction rule derived from the management information system of an HMO: a preliminary study. AB - BACKGROUND: Somatization is a common, costly problem with great morbidity, but there has been no effective screening method to identify these patients and target them for treatment. OBJECTIVES: We tested a hypothesis that we could identify high utilizing somatizing patients from a management information system (MIS) by total number of visits and what we termed "somatization potential," the percentage of visits for which ICD-9 primary diagnosis codes represented disorders in the musculoskeletal, nervous, or gastrointestinal systems or ill defined complaints. METHODS: We identified 883 high users from the MIS of a large staff model HMO as those having six or more visits during the year studied (65th percentile). A physician rater, without knowledge of hypotheses and predictors, then reviewed the medical records of these patients and identified somatizing patients (n = 122) and nonsomatizing patients (n = 761). In two-thirds of the population (the derivation set), we used logistic regression to refine our hypothesis and identify predictors of somatization available from the MIS: demographic data, all medical encounters, and primary diagnoses made by usual care physicians (ICD-9 codes). We then tested our prediction model in the remaining one-third of the population (the validation set) to validate its usefulness. RESULTS: The derivation set contained the following significant correlates of somatization: gender, total number of visits, and percent of visits with somatization potential. The c-statistic, equivalent to the area under the ROC curve, was 0.90. In the validation set, the explanatory power was less with a still impressive c-statistic of 0.78. A predicted probability of 0.04 identified almost all somatizers, whereas a predicted probability of 0.40 identified about half of all somatizers but produced few false positives. CONCLUSIONS: We have developed and validated a prediction model from the MIS that helps to distinguish chronic somatizing patients from other high utilizing patients. Our method requires corroboration but carries the promise of providing clinicians and health plan directors with an inexpensive, simple approach for identifying the common somatizing patient and, in turn, targeting them for treatment. The screener does not require clinicians' time. PMID- 11502955 TI - Relative impact of patient and clinic factors on adherence to primary care preventive service guidelines: an exploratory study. AB - BACKGROUND: Preventive care service use is commonly compared across health plans, clinics, or individual providers, yet little is known about the influence of the clinic versus patient factors on utilization of these services. OBJECTIVES: To measure the relative influence of the facility (clinic) versus patient factors (demographic, behavioral and functional characteristics) on patients' utilization of mammography, Pap smears, cholesterol screening, and retinal exams for those with diabetes. RESEARCH DESIGN: Retrospective analysis, using administrative and patient survey data. SUBJECTS: Enrollees in 2 University-based clinics and a county hospital-based clinic serving a predominantly low-income population with limited access to health care. Eligibility for cervical cancer screening, screening mammography, cholesterol screening, or annual retinal exam (diabetes) was defined by age, sex, and diagnosis. MEASURES: Multivariate models, one using readily available administrative data, and another using detailed health status and behavior data gathered from a clinics-wide survey. RESULTS: Unadjusted screening rates for three of four procedures were significantly and substantially lower at the county hospital based clinic than the two University-based clinics. After adjusting for patient characteristics, utilization of three screening services at the county hospital remained significantly below the University-based clinics (Odds Ratios [95% CI]: mammogram 0.15 [0.06-0.35]; Pap smear 0.32 [0.21 0.50]; cholesterol 0.19 [0.09-0.38]; diabetes retinal exam10.68 [0.93-3.01]). The models with detailed survey data performed only marginally better than the models using only administrative data. CONCLUSIONS: Patient characteristics were much less important than the clinic for predicting whether patients received primary care preventive services. Our results suggest that case mix adjustment is unlikely to explain away discrepancies in performance between clinics or provider groups. PMID- 11502956 TI - Children's health care use: a prospective investigation of factors related to care-seeking. AB - OBJECTIVES: To determine the best predictors of the amount of children's health care use. RESEARCH DESIGN: Child health, psychosocial, and family status variables were collected. Families were then followed prospectively for 2 years to gather health care use data. Multivariate regression analysis was used to determine factors related to volume of child health care use. SUBJECTS: 367 mothers and children ages 5 to 11 years continuously enrolled in a staff model HMO. MEASURES: Child health care visits obtained from a computerized database comprised the dependent variable. Independent variables were organized into a 5 component framework including: Demographic Characteristics; Family Characteristics; Child Health and Prior Health Care Use; Child Behavior and Mental Health; and Mothers' Mental Health and Health Care Use. RESULTS: The volume of a child's past health care use was the best predictor of future health care use, with the presence of past acute recurring illnesses, child pain and mother's retrospective health care use also serving as significant predictors in the model. Analysis of a second model was conducted omitting children's past use of health care. In this model the mother's worry about child health was the best predictor of use, with child health and child and maternal psychosocial variables significantly contributing to explained variance in the model. CONCLUSIONS: This study supports prior research indicating past use is the best predictor of future health care use. In addition, the study suggests that maternal perceptions of child health and maternal emotional functioning influence the decision-making process involved in seeking health care on behalf of children. Effective management of pediatric health care use needs to address broader needs of the child and family beyond solely the child's health, most notably maternal functioning. PMID- 11502957 TI - Heterogeneity of risk in a managed home health care population. AB - OBJECTIVE: To examine the distribution of risk and the correlation between risks in a home care population with regard to several important adverse outcomes. BACKGROUND: Researchers and policy makers have long recognized the heterogeneity of home care populations. Most research in this area focuses on identifying predictors of adverse outcomes. The degree of the heterogeneity of risks is much more poorly understood. Yet understanding the degree of risk heterogeneity at the population level is important because it has implications for the extent to which the level of care should vary among recipients. STUDY SETTING: Patients enrolled in the Arizona Health Care Cost Containment System (AHCCCS) program, between the December 1992 and April 1998. OUTCOME MEASURES: Estimating the risk for nursing home placement, hospitalization, death, and functional decline. METHODS: Estimating discrete time hazard models. From these models the predicted risk for each outcome is estimated and the distribution and correlation of predicted risks is examined. Model fit is assessed through split sample techniques and by examining the ratio of predicted to actual outcomes for selected sub-groups. RESULTS: The estimates reveal a wide variation in predicted risk. The ratio of predicted risk at the 90th percentile relative to the 10th percentile ranges from 4.99 for nursing home admission to 6.65 for hospitalization. The distributions of predicted risks are all skewed, particularly the distributions for death and nursing home admission. Predicted nursing home risk is highly correlated with the predicted risk for death (rho = 0.71). The predicted risk for hospitalization is not strongly correlated with the predicted risk for either death or nursing home admission. CONCLUSION: The wide variation in risk among home care patients suggests that efficient allocation of resources would require variation in spending and targeting of services based on patient characteristics. Greater research regarding the effectiveness of home care for different sub-populations is called for. PMID- 11502958 TI - Length of stay as a source of bias in comparing performance in VA and private sector facilities: lessons learned from a regional evaluation of intensive care outcomes. AB - OBJECTIVES: Compare intensive care unit (ICU) mortality and length of stay (LOS) in a VA hospital and private sector hospitals and examine the impact of hospital utilization on mortality comparisons. RESEARCH DESIGN: Retrospective cohort study. SUBJECTS: Consecutive ICU admissions to a VA hospital (n = 1,142) and 27 private sector hospitals (n = 51,249) serving the same health care market in 1994 to 1995. MEASURES: Mortality and ICU LOS were adjusted for severity of illness using a validated method that considers physiologic data from the first 24 hours of ICU admission. Mortality comparisons were made using two different multivariable techniques. RESULTS: Unadjusted in-hospital mortality was higher in VA patients (14.5% vs. 12.0%; P = 0.01), as was hospital (28.3 vs. 11.3 days; P <0.001) and ICU (4.3 vs. 3.9 days; P <0.001) LOS. Using logistic regression to adjust for severity, the odds of death was similar in VA patients, relative to private sector patients (OR 1.16, 95% CI 0.93-1.44; P = 0.18). However, a higher proportion of VA deaths occurred after 21 hospital days (33% vs. 13%; P <0.001). Using proportional hazards regression and censoring patients at hospital discharge, the risk for death was lower in VA patients (hazard ratio 0.70; 95% CI 0.59-0.82; P <0.001). After adjusting for severity, differences in ICU LOS were no longer significant (P = 0.19). CONCLUSIONS: Severity-adjusted mortality in ICU patients was lower in a VA hospital than in private sector hospitals in the same health care market, based on proportional hazards regression. This finding differed from logistic regression analysis, in which mortality was similar, suggesting that comparisons of hospital mortality between systems with different hospital utilization patterns may be biased if LOS is not considered. If generalizable to other markets, our findings further suggest that ICU outcomes are at least similar in VA hospitals. PMID- 11502959 TI - Expectations regarding aging among older adults and physicians who care for older adults. AB - BACKGROUND: Understanding older adults' expectations regarding aging is important for both clinicians and policy-makers. OBJECTIVES: 1) To identify the content for a survey to measure expectations regarding aging; 2) to qualitatively compare older adults' and physicians' expectations regarding aging. SETTING: Three senior centers and one university-based internal medicine faculty practice. PARTICIPANTS: Forty-nine adults (mean age 78 years); 11 primary care clinicians (mean age 37 years). MEASUREMENTS: A facilitator conducted five focus groups of older adults and two of physicians using a standardized script designed to elicit expectations regarding aging. Qualitative analysis by two independent reviewers identified domains of expectations, with a 3rd reviewer used to resolve discrepancies. A corresponding coding scheme was applied to each line of the transcripts. Content and frequency of expectations regarding aging and beliefs regarding care seeking were examined and compared. RESULTS: Content analysis identified 26 domains of expectations regarding aging. Each of the seven most frequently mentioned domains of expectations was mentioned by at least 50% of participants. Of 760 unique statements coded, the most frequently described domains in both the older adult and physician groups were physical function, cognitive function, social function, pain, and sexual function. Older adults differed from physicians by describing five mental-health related domains: anxiety, emotional-well-being, happiness, sleep, and length of life/death. CONCLUSIONS: Using focus groups of older adults and physicians, we identified consistent content for a closed-ended patient-centered survey to measure expectations regarding aging. Further study should determine whether physicians address mental health aspects of aging valued by older persons. PMID- 11502960 TI - Donor or recipient origin of posttransplantation lymphoproliferative disorders: the traces of the Epstein-Barr virus. PMID- 11502961 TI - Should catecholamines be given to brain dead donors? PMID- 11502962 TI - Viral serine proteinase inhibitor (SERP-1) effectively decreases the incidence of graft vasculopathy in heterotopic heart allografts. AB - BACKGROUND: Graft vascular disease (GVD) is the most common cause of late graft failure in solid organ transplantation. Recent studies have shown good efficacy of a novel nontoxic viral-derived serine proteinase inhibitor (SERP-1) in preventing postangioplasty restenosis. The current study was designed to test whether short-term treatment with SERP-1 was effective in reducing the incidence of GVD in a solid organ transplant. METHODS: Piebald-Virol-Glaxo (PVG) donor hearts were transplanted into August-Copenhagen-Irish (ACI) recipients and observed for 90 days. All recipients (n=60) were treated with microemulsion cyclosporine (CsA) 7.5 mg/kg per gavage from day 0 to day 9 and randomized into 4 groups. SERP-1 was given intravenously. Group I received CsA monotherapy; group II, CsA+SERP-1 1 ng/g (postoperative days 0-9); group III, CsA+SERP-1 10 ng/g (postoperative days 0-9); and group IV, CsA+SERP-1 10 ng/g (postoperative days 0 9, 30, and 60). Graft viability was monitored by palpation, and GVD was assessed by morphometry. RESULTS: Two animals in group I rejected their allografts on postoperative days 7 and 14, 1 animal in group II rejected the allograft (postoperative day 31), and none in group III and IV rejected the allografts. At 90 days postoperative, 23.8% of all coronary vessels showed evidence of GVD in group I, 18.4% in group II, 12.9% in group III, and 11.8% in group IV. The difference in incidence of GVD was significant between groups I and III (P<0.05) and groups I and IV (P<0.05). Treatment with SERP-1 was well tolerated, and all animals regained weight quickly postsurgery. CONCLUSIONS: Treatment of allograft recipients with SERP-1 in combination with CsA early after transplantation significantly decreases the incidence of GVD when compared to grafts treated with only CsA. These results demonstrate the clinical potential for this novel serine protease inhibitor to prevent GVD in solid organ transplantation. PMID- 11502963 TI - Suppression mediated by anergic CD4+ T cells requires stimulation by MHC-peptide complexes and can be induced in the presence of costimulation. AB - BACKGROUND: In this study, we have investigated the mechanisms involved in both the induction of suppressive anergy, the stability of the anergy induced, and the possible mechanisms by which the response of immunocompetent CD4+ T cells are suppressed. METHODS: We used immobilized anti-CD3 monoclonal antibody (mAb) to induce anergy in T helper (Th) 1 and Th0 cells reactive with MHC class II molecule H2 I-Ab. RESULTS: We observed that suppressive anergy was induced independently of costimulation in Th0 but not Th1 cells. Although the anergic and suppressive states of Th0 cells were stable in the presence of exogenous interleukin-2, this was not the case for Th1 cells. No evidence for linked epitope suppression was observed for any of the I-Ab reactive cells investigated. Neither anergy nor suppression was observed in Th0 cells upon restimulation with anti-CD3 in the presence of syngeneic antigen-presenting cells (APCs). However, anergy but not suppression was observed in co-cultures restimulated with anti-T cell antigen receptor (TCR) mAbs/syngeneic APCs and suppression could be restored by the addition of I-Ab+ APCs. CONCLUSIONS: Overall, these data suggested that the MHC-peptide complex recognized by the Th0 cells was required for suppression of the response of immunocompetent cells. We propose that suppression is mediated either by down-modulation of the MHC-peptide complex recognized by the anergic T cells or that a molecule specific to the MHC-peptide/TCR interaction facilitates negative regulation by APC:T or T:T interactions. PMID- 11502964 TI - Induction therapy with monoclonal antibodies specific for CD80 and CD86 delays the onset of acute renal allograft rejection in non-human primates. AB - CD80 and CD86 (also known as B7-1 and B7-2, respectively) are both ligands for the T cell costimulatory receptors CD28 and CD152. Both CD80 and CD86 mediate T cell costimulation, and as such, have been studied for their role in promoting allograft rejection. In this study we demonstrate that administering monoclonal antibodies specific for these B7 ligands can delay the onset of acute renal allograft rejection in rhesus monkeys. The most durable effect results from simultaneous administration of both anti-B7 antibodies. The mechanism of action does not involve global depletion of T or B cells. Despite in vitro and in vivo evidence demonstrating the effectiveness of the anti-B7 antibodies in suppressing T cell responsiveness to alloantigen, their use does not result in durable tolerance. Prolonged therapy with murine anti-B7 antibodies is limited by the development of neutralizing antibodies, but that problem was avoided when humanized anti-B7 reagents are used. Most animals develop rejection and an alloantibody response although still on antibody therapy and before the development of a neutralizing antibody response. Anti-B7 antibody therapy may have use as an adjunctive agent for clinical allotransplantation, but using the dosing regimens we used, is not a tolerizing therapy in this non-human primate model. PMID- 11502965 TI - Selective attenuation of acute lung ventilatory injury by methylene blue after liver ischemia-reperfusion: a drug response study in an isolated perfused double organ model. AB - BACKGROUND: Liver transplantation-related ischemia-reperfusion (IR) is associated with the generation of stress oxidants that can spread damage remotely. Methylene blue (MB) had been shown to reduce lung neutrophils sequestration after in vivo intestinal IR and to have a dose-dependent potential for abrogating oxidant induced ex vivo aortal ring reperfusion injury after liver IR. We now investigated MB's dose-dependent capabilities in preventing acute lung injury after the same liver IR. METHODS: Wistar rat livers (eight replicates/group) were perfused (control) with modified Krebs-Henseleit solution or put globally in no flow (IR) conditions for 2 hr. Separately prepared lungs were then paired with livers and "reperfused" (15 min) together. The livers were then removed, and the lungs were left to recirculate alone with the accumulated Krebs for 45 min. Three additional control and three IR groups were reperfused with Krebs containing 20, 40, or 60 mg/kg MB at concentrations of 42, 86, or 128 microM. RESULTS: All IR livers had hepatocellular and biochemical abnormalities compared with normal functions in the controls. Liver IR was associated with a 50%-75% increase in lung ventilation and perfusion pressures, vascular resistance and decreased compliance, and abnormal bronchoalveolar lavage (BAL) volume and content. Adding 42 and 86 microM MB selectively maintained normal the vascular parameters, intra experimental lung weight gain, BAL indices, and wet-to-dry ratios. MB128 microM but not 42 or 86 microM best prevented IR-induced deterioration in lung ventilatory pressure and compliance. CONCLUSIONS: MB selectively affords maintenance of normal lung ventilatory versus vascular measures after liver ischemia-reperfusion. Its proposed differential mechanism of action is discussed. PMID- 11502966 TI - Functional metabolic characteristics of intact pig livers during prolonged extracorporeal perfusion: potential for a unique biological liver-assist device. AB - BACKGROUND: The clinical development of liver-support devices based on perfusion of either pig hepatocytes cartridges or whole pig livers has been hampered by the ability to use sufficient liver cell mass to provide adequate metabolic support, limited perfusion times, and the potential for patient exposure to pig zoonotic diseases. METHODS: We designed an original system in which an isolated intact pig liver was perfused extracorporeally under physiological conditions in a closed loop circuit with allogeneic pig blood and constant monitoring of major physiological and functional parameters. The perfusion circuit further included an interface membrane to provide for separation of patient and liver perfusion circulation. RESULTS: Prolonged (6-21 hr) liver perfusion did not produce significant liver damage as reflected by modest rises in the levels of the serum transaminases, stability of main biochemical parameters (including potassium), and the maintenance of normal cellular morphology. Optimal liver function was documented as measured by lactate consumption, control of glycemia, and the results of clotting studies and functional assays. The perfused liver cleared 82% and 79% of peak bilirubin and ammonia concentrations with clearing kinetics identical throughout perfusion. Indocyanine green clearance was identical to that observed in the living donor before explant surgery. CONCLUSIONS: In conclusion, the extracorporeal pig liver perfusion apparatus described here allows optimal pig liver function for prolonged periods of time. The microporous membrane to provide separation of donor organ and recipient and the high level of functional activity suggest that this form of liver metabolic support may have important clinical applications. PMID- 11502967 TI - Protective effects of ischemic preconditioning on the cold-preserved liver are tyrosine kinase dependent. AB - BACKGROUND: Little data exist regarding the use of ischemic preconditioning before sustained hepatic cold storage. We hypothesized that ischemic preconditioning protects hepatic grafts via a tyrosine kinase-dependent pathway. METHODS: Six porcine livers underwent routine harvest (control). Five other livers underwent 15 min of in situ ischemia followed by 15 min of reflow before harvest (ischemic preconditioning). Another five livers were pretreated with a tyrosine kinase inhibitor (genistein) before preconditioning. Upon reperfusion and after 2 hours of cold storage, graft function, graft circulatory impairment, and markers of cellular damage were analyzed. Tissue cytoplasmic extracts were analyzed for tyrosine phosphorylation with Western blot. Significance was determined with t tests. RESULTS: Ischemic-preconditioned grafts demonstrated enhanced bile production, augmented responses to a bile acid challenge, and elevated O2 consumption (P<0.05) compared to controls. Also, preconditioned grafts demonstrated improved hepatic tissue blood flow and decreased hepatic vascular resistance (P<0.005) compared to controls. Endothelial cell preservation (factor VIII immunostain) was improved in preconditioned graft biopsies compared to controls. With genistein pretreatment, all observed improvements returned to control levels. Analysis of cytoplasmic extracts demonstrated an increase in tyrosine phosphorylation before cold ischemia in preconditioned grafts only, but not in control or genistein-pretreated grafts. CONCLUSIONS: The data indicate that ischemic preconditioning protects the liver from sustained cold ischemia and that tyrosine kinases are involved in preconditioning responses. PMID- 11502968 TI - The role of different immunosuppression in the long-term histological outcome of HCV reinfection after liver transplantation for HCV cirrhosis. AB - BACKGROUND: The effect of the type of immunosuppression on the course of posttransplant hepatitis C virus (HCV) infection is unclear. The aim of this study was to evaluate the histological outcome of posttransplant HCV infection with respect to initial immunosuppressive therapy in a cohort of 59 of 65 HCV positive transplant patients who survived at least 12 months. METHODS: Initial immunosuppressive therapy was triple (cyclosporine or tacrolimus and azathioprine and prednisolone) in 41, double (cyclosporine and prednisolone) in 5, and single (cyclosporine or tacrolimus) in 13 patients. There was blinded histological evaluation, based on necroinflammatory activity (grading score:0-18) and fibrosis (staging score: 0-6). The median histological follow-up was 36 (12-72) months. RESULTS: In the last liver biopsy, high necroinflammatory activity indicating chronic hepatitis (grading score > or =4) was found in 42 (71%) and severe fibrosis or cirrhosis (staging score > or =4) in 18 (30.5%) patients. High necroinflammatory activity was associated significantly with absence of pretransplant alcohol abuse (P=0.01) and relatively with occurrence of posttransplant acute lobular hepatitis C (P=0.055). Development of severe fibrosis or cirrhosis was significantly associated only with the type of initial immunosuppressive therapy. In particular, severe fibrosis or cirrhosis developed significantly more frequently in patients treated with triple or double (17/46 or 37%) than with single initial immunosuppressive therapy (1/13 or 7.7%) (adjusted for biopsy time: P=0.045). CONCLUSIONS: Severe fibrosis or cirrhosis appears to develop in 30% of HCV transplant patients in a median of 3 years and to be associated with heavier initial immunosuppression. PMID- 11502969 TI - Reversal of steroid- and anti-lymphocyte antibody-resistant rejection using intravenous immunoglobulin (IVIG) in renal transplant recipients. AB - BACKGROUND: Despite the recent advances in immunosuppression, steroid-resistant rejection remains a difficult problem in renal transplant recipients. METHODS: We reviewed our experience with i.v. immunoglobulin (IVIG) in the treatment of steroid- and antilymphocyte antibody-resistant rejection in renal transplant patients. Between September 1996 and March 1999, 17 patients were treated with IVIG to reverse steroid- or antilymphocyte antibody-resistant rejection. A total of 2 g/kg of IVIG was administered to patients during each treatment course. RESULTS: With a mean follow-up of 21.5+/-9.5 months from the time of IVIG administration, patient and graft survival rates were 94% (16/17) and 71% (12/17), respectively. The baseline mean serum creatinine level prior to rejection was 2.2+/-0.7 mg/dl and peaked at 3.3+/-1.1 mg/dl at the time of the diagnosis of refractory rejection. IVIG therapy was associated with a fall in the mean creatinine to 2.8+/-1.1 mg/dl. The most recent serum creatinine in patients with functioning grafts was 2.8+/-1.6 mg/dl. In 82% of allograft biopsies after IVIG, reversal or reduction in the severity of rejection was demonstrated. In addition, IVIG therapy rescued three of four patients with antilymphocyte antibody-resistant rejection. CONCLUSIONS: IVIG rescue therapy for steroid- or antilymphocyte antibody-resistant rejection is associated with resolution or improvement of rejection severity, stable renal function, and reasonable graft survival. PMID- 11502970 TI - Measurement of mycophenolate mofetil effect in transplant recipients. AB - BACKGROUND: Immunosuppression involves the nature of the immunosuppressive agents and individual differences in patient factors. We investigated whether the effect of mycophenolate mofetil (MMF) is measurable using an in vitro measure of immunocompetence and related its effects to cyclosporine (CsA) in vitro. METHODS: Liver or kidney transplant recipients receiving prednisone; CsA or tacrolimus; and MMF, azathioprine (AZA), or neither, were studied. Immunocompetence was assessed by one-way mixed lymphocyte culture using patients' peripheral blood leukocytes (PBL) and three validated stimulators. The effect of immunosuppressive agents added in vitro on normal PBL stimulation by Staphylococcus enterotoxin B was determined by the carboxyfluorescein diacetate succinimidyl ester measurement of division. RESULTS: Patients receiving MMF had an average immunocompetence level of 12+/-23, compared with 39.7+/-65 and 25.5+/-42 for those receiving AZA or neither AZA nor MMF, respectively. Thus, there was an approximately 80% suppression of the response in the MMF group. Assessment of normal cell division revealed that CsA allowed multiple cell generations but suppressed the numbers of cells in each, whereas MMF blocked proliferation into subsequent generations. Addition of clinically relevant levels of mycophenolic acid, the active agent for MMF, added to more moderate levels of CsA, was required to achieve greater than 80% suppression, consistent with the degree of immunocompetence depression measured in patients. CONCLUSIONS: These data provide the novel finding that the effect of MMF treatment on patients is measurable in their PBL as decreased immunocompetence in vitro. The effect of MMF on normal PBL approximates the 80% inhibition that we found in patients. Moreover, the effect of MMF on cell division provides a rationale for the superior effectiveness of regimens including MMF. PMID- 11502971 TI - Primary hyperoxaluria type 1: improved outcome with timely liver transplantation: a single-center report of 36 children. AB - BACKGROUND: The appropriate use of liver transplantation in children with type-1 primary hyperoxaluria (PH-1) is not well established. We reviewed our experience with 36 children with PH-1, including 12 who underwent liver transplantation. PATIENTS AND METHODS: From 1989-1998, 36 children from 10 families in northern Israel were diagnosed with PH-1. Eight children presented with renal failure; seven of these eight had the severe infantile form of the disease. One child was treated with kidney transplantation alone. Combined liver-kidney transplantation has been performed in nine children and preemptive liver transplantation in three children. A review of the patients' charts for the following parameters was performed: age, clinical signs, and renal sonographic findings at diagnosis, age at onset of dialysis, and current status. Type of transplant, pre- and posttransplant urine oxalate excretion, current renal function, survival, and complications were recorded in liver recipients. RESULTS: Of the 23 nontransplanted children, 9 died of complications related to severe systemic oxalosis and 14 are alive (mean follow-up, 7.4 years), including 2 who are candidates for transplantation. The child who underwent only kidney transplantation died of unrelated causes. Of the 12 liver recipients, 2 died within the first 3 months posttransplant and another child underwent retransplantation due to hepatic arterial thrombosis. At intervals after transplant ranging from 6-54 months, 10 recipients are alive (7 of the 9 recipients of combined liver-kidney transplants and all 3 recipients of preemptive liver transplants). Mean GFR in the 10 survivors is 77 ml/min/m2. In 9 of these 10, daily urinary oxalate excretion normalized. Renal function has improved (mean GFR 86 vs. 58 ml/min/m2) but renal oxalate deposits remain in the three recipients of isolated liver grafts. CONCLUSIONS: Our decade-long experience with children with PH-1 supports strategies for early diagnosis and timely liver transplantation. Preemptive isolated liver transplantation should be considered in children who develop the disease during infancy or in those with slowly progressive disease when significant symptoms develop. Combined liver kidney transplantation is suggested for children with end-stage renal disease. PMID- 11502972 TI - Effect of Bw4 and Bw6 epitope mismatches on antibody production, acute and chronic rejection, and graft survival in renal allografts. AB - BACKGROUND: Highly sensitized patients often have antibodies directed against the HLA Bw4 and Bw6 epitopes. Because of the high frequency of these epitopes, when present, these antibodies result in a high incidence of positive cross-matches. We sought to determine whether antibodies specific for Bw4 or Bw6 affected renal allograft outcome. METHODS: The effect of mismatches for the HLA class I public epitopes, Bw4 and Bw6, was examined in 72 recipients of one haplotype matched recipients of living, related donor renal allografts selected to control for degree of HLA mismatch. Analysis of the production of HLA-specific antibody was performed for 180 recipients of failed cadaveric allografts by complement dependent cytotoxicity tests and by an enzyme-linked immunoadsorbent assay (ELISA). RESULTS: No significant difference was observed in the incidence of acute rejection, number of rejection episodes or 1-year allograft survival among Bw4/6 matched versus mismatched recipients of one haplotype matched allografts. Additionally, no significant difference in the development of chronic allograft nephropathy was noted among 56 recipients followed long-term (> or =3 years). In the recipients of failed cadaveric transplants, Bw4/6 mismatching was associated with the frequency and magnitude of production of HLA-specific antibody. However, the panel reactive antibodies correlated with the number of HLA-A and -B mismatches, and there was no additional impact of Bw4/6 mismatching. IgG, HLA specific antibodies were found to be significantly increased among patients homozygous for Bw4 or Bw6, whether or not there was a Bw4/6 mismatch. CONCLUSIONS: Mismatching for Bw4 or Bw6 does not confer any independent, increased risk for humoral sensitization or renal allograft failure. PMID- 11502973 TI - Proteinuria after renal transplantation affects not only graft survival but also patient survival. AB - BACKGROUND: Proteinuria is associated with an increased risk of renal failure. Moreover, proteinuria is associated with an increased death risk in patients with diabetes mellitus or hypertension and even in the general population. METHODS: One year after renal transplantation, we studied the influence of the presence of proteinuria on the risk of either graft failure or death in all 722 recipients of a kidney graft in our center who survived at least 1 year with a functioning graft. Proteinuria was analyzed both as a categorical variable (presence versus absence) and as a continuous variable (quantification of 24 hr urine). Other variables included in this analysis were: donor/recipient age and gender, original disease, race, number of HLA-A and HLA-B mismatches, previous transplants, postmortal or living related transplantation, and transplantation year. At 1 year after transplantation, we included: proteinuria, serum cholesterol, serum creatinine, blood pressure, and the use of antihypertensive medication. RESULTS: In the Cox proportional hazards analysis, proteinuria at 1 year after transplantation (both as a categorical and continuous variable) was an important and independent variable influencing all endpoints. The influence of proteinuria as a categorical variable on graft failure censored for death showed no interaction with any of the other variables. There was an adverse effect of the presence of proteinuria on the graft failure rate (RR=2.03). The influence of proteinuria as a continuous variable showed interaction with original disease. The presence of glomerulonephritis, hypertension, and systemic diseases as the original disease significantly increased the risk of graft failure with an increasing amount of proteinuria at 1 year. The influence of proteinuria as a categorical variable on the rate ratio for patient failure was significant, and there was no interaction with any of the other significant variables (RR=1.98). The death risk was almost twice as high for patients with proteinuria at 1 year compared with patients without proteinuria. The influence of proteinuria as a continuous variable was also significant and also without interaction with other variables. The death risk increased with increasing amounts of proteinuria at 1 year. Both the risks for cardiovascular and for noncardiovascular death were increased. CONCLUSION: Proteinuria after renal transplantation increases both the risk for graft failure and the risk for death. PMID- 11502974 TI - No evidence of accelerated loss of kidney function in living kidney donors: results from a cross-sectional follow-up. AB - BACKGROUND: There is a lack of kidneys available for kidney transplantation, and living donors are increasingly used. It is important to examine the possible long term adverse affect on the renal function and blood pressure of the donors. METHODS: We have made a comprehensive follow-up of all living kidney donors at our center from 1964 to 1995. Of 402 donors still alive, we were able to get information about serum creatinine, urinary proteins, and blood cells in urine using reagent strips, and blood pressure from 87%. The glomerular filtration rate (GFR) was estimated using a formula and was measured with Iohexol clearance in 43 of the donors. Individual data on GFR and the prevalence of hypertension were compared with the age- and gender-expected values. RESULTS: The mean age of the examined donors was 61 years (SD:13) at follow-up, and the time since donation was 12 years (SD:8). The average estimated GFR was 72% (SD:18) of the age predicted value. The ratio of the estimated to the predicted GFR showed no correlation to the time since donation, indicating that there is no accelerated loss of renal function after donation. GFR below 30 ml/min was found in five donors. No donor died in uremia or had dialysis treatment before death. However, three donors developed renal disease, and one was in dialysis treatment. In two of these cases, hereditary factors were possibly involved. Hypertension was present in 38% of the donors but the age-adjusted prevalence of hypertension among donors was not higher than in the general population. Significant proteinuria (> or =1.0 g/L) was found in 3% and slight proteinuria (<1.0 g/L) in 9% of the donors. Proteinuria was associated with hypertension and a lower GFR. CONCLUSIONS: On average, the remaining renal function of kidney donors did not deteriorate more rapidly than what may be expected from ageing. However one-third of the female and half of the male donors developed hypertension and, approximately, 10% displayed proteinuria. Nevertheless, our study supports the continued use of living kidney donors if strict criteria are used for acceptance. PMID- 11502975 TI - Weaning of immunosuppression in living donor liver transplant recipients. AB - BACKGROUND: Some reported studies have indicated the possibility of immunosuppression withdrawal in cadaveric liver transplantation. The aim of this study was to evaluate the possibility and feasibility of weaning living donor liver transplant recipients from immunosuppression. METHODS: From June of 1990 to October of 1999, 63 patients were considered to be weaned from immunosuppression. They consisted of 26 electively weaned patients and 37 either forcibly or incidentally weaned patients (nonelective weaning) due to various causes but mainly due to infection. Regarding elective weaning, we gradually reduced the frequency of tacrolimus administration for patients who survived more than 2 years after transplantation, maintained a good graft function, and had no rejection episodes in the preceding 12 months. The frequency of administration was reduced from the conventional b.i.d. until the start of weaning to q.d., 4 times a week, 3 times a week, twice a week, once a week, twice a month, once a month, and finally, the patients were completely weaned off with each weaning period lasting from 3 to 6 months. The reduction method of nonelective weaning depended on the clinical course of each individual case. When the patients were clinically diagnosed to develop rejection during weaning, then such patients were treated by a reintroduction of tacrolimus or an additional steroid bolus when indicated. RESULTS: Twenty-four patients (38.1%) achieved a complete withdrawal of tacrolimus with a median drug-free period of 23.5 months (range, 3-69 months). Twenty-three patients (36.5%) are still being weaned at various stages. Sixteen patients (25.4%) encountered rejection while weaning at median period of 9.5 months (range, 1-63 months) from the start of weaning. All 16 were easily treated with the reintroduction of tacrolimus or additional steroid bolus therapy. CONCLUSIONS: We were able to achieve a complete withdrawal of immunosuppression in some selected patients. Although the mechanism of graft acceptance in these patients has yet to be elucidated, we believe that a majority of long-term patients undergoing living donor liver transplantation may, thus, be potential candidates to be successfully weaned from immunosuppression. PMID- 11502976 TI - Effects of catecholamine application to brain-dead donors on graft survival in solid organ transplantation. AB - BACKGROUND: In a recent single-center study, donor use of catecholamines was identified to reduce kidney allograft rejection. This study investigates the effects of donor employment of adrenergic agents on graft survival in a large data base, including liver and heart transplants. METHODS: The study was based on the registry of the Eurotransplant International Foundation including 2415 kidney, 755 liver, and 720 heart transplants performed between January 1 and December 31, 1993. A total of 1742 donor record forms referring to the cadaveric donor activities in 1993 were systematically reviewed with regard to employment of adrenergic agents. Catecholamine use was simply coded dichotomously and divided into three strata according to zero, single, and combined application. Multivariate Cox regression including age, gender, cause of brain death, cold ischemia, HLA-mismatching, number of previous transplants, and urgency in liver transplants was applied for statistical analysis. RESULTS: Donor employment of catecholamines was associated with increased 4-year graft survival after kidney transplantation (hazard ratio [HR], 0.85; 95% confidence interval [95% CI], 0.74 0.98). The benefit is conferred in a dose-dependent manner and compares in quantitative terms with prospective HLA matching on class I and class II antigens (HR, 0.90; 95% CI, 0.84-0.97). Use of norepinephrine was predictive of initial nonfunction after heart transplantation (HR, 1.66; 95% CI, 1.14-2.43), but did not compromise liver grafts (HR, 0.94; 95% CI, 0.67-1.32). CONCLUSIONS: Optimizing the management of brain-dead organ donors, including the possibility of selective administration of adrenergic agents, may provide a major benefit on graft survival without adverse side effects for the recipients. Further investigation on best use of adrenergic drugs, optimum dosage, and duration is warranted. PMID- 11502977 TI - Studies of Pediatric Liver Transplantation (SPLIT): year 2000 outcomes. AB - BACKGROUND: Initiated in 1995, the Studies of Pediatric Liver Transplantation (SPLIT) registry database is a cooperative research network of pediatric transplantation centers in the United States and Canada. The primary objectives are to characterize and follow trends in transplant indications, transplantation techniques, and outcomes (e.g., patient/graft survival, rejection, growth parameters, and immunosuppressive therapy.) METHODS: As of June 15, 2000, 29 centers registered 1144 patients, 640 of whom received their first liver-only transplant while registered in SPLIT. Patients are followed every 6 months for 2 years and yearly thereafter. Data are submitted to a central coordinating center. RESULTS: One/two-year patient survival and graft loss estimates are 0.85/0.82 and 0.77/0.72, respectively. Risk factors for death include: in ICU at transplant (relative risk (RR)=2.63, P<0.05) and height/weight deficits of two or more standard deviations (RR=1.67, P<0.05). Risk factors for graft loss include: in ICU at transplant (RR=1.77, P<0.05) and receiving a cadaveric split organ compared with a whole organ (RR=2.3, P<0.05). The percentage of patients diagnosed with hepatic a. and portal v. thrombosis were 9.7% and 7%, respectively; 15% had biliary complications within 30 days. At least one re operation was required in 45%. One/two-year rejection probability estimates are 0.60/0.66. Tacrolimus, as primary therapy posttransplant, reduces first rejection risk (RR=0.70, P<0.05). Eighty-nine percent of school-aged children are in school full-time, 18 months posttransplant. CONCLUSIONS: This report provides one of the first descriptions of characteristics and clinical courses of a multicenter pediatric transplant population. Observations are subject to patient selection biases but are useful for generating hypothesis for future studies. PMID- 11502978 TI - Candida fasciitis following renal transplantation. AB - BACKGROUND: We describe a rare case of necrotizing fasciitis involving Candida albicans, an organism that has been reported to have a minimal potential for invasive soft tissue infection. In this case, immunosuppression, chronic renal failure, and a history of diabetes mellitus were predisposing factors. METHODS: The medical record and histopathologic material were examined. The clinical literature was reviewed for previous cases of C albicans necrotizing fasciitis. RESULTS: A review of the literature showed that in solid organ transplant recipients, localized fungal soft tissue infection is infrequent, with only 35 cases reported between 1974 and 1992. Necrotizing fasciitis caused by C albicans is extremely rare in the modern era of solid organ transplantation. CONCLUSIONS: The management of transplant patients at risk for invasive fungal infection warrants a high index of suspicion for fungal necrotizing fasciitis in the setting of wound infection and merits a thorough investigation for atypical pathogens. PMID- 11502979 TI - The role of the allograft in the induction of donor-specific T cell hyporesponsiveness. AB - BACKGROUND: With adequate immunosuppression the majority of renal allografts are accepted, despite the exceptional vigour of the T cell alloimmune response. Previous work from this laboratory has demonstrated that this is accompanied by significant reductions in the precursor frequencies of anti-donor T cells. We have also shown that parenchymal cells are tolerogenic in vitro. We propose that the reduction in T cell frequencies may be due to the interaction between circulating T cells and potentially tolerogenic graft parenchymal cells. Primed/memory T cells (CD45RO+) are the only subset capable of reaching the allograft and therefore we would predict that T cell hyporesponsiveness would develop predominantly in the CD45RO+ subset due to their trafficking properties. METHODS: Frequencies of IL-2 secreting CD45RA+ and CD45RO+ CD4+ T cells in response to donor and third party stimulator cells were estimated in a series of renal transplant recipients, both before and after transplantation. RESULTS: There were highly significant reductions in the frequencies of donor-specific CD4+CD45RO+ T cells, when adjusted to control for the generalised effects of immunosuppression. There were no significant alterations in the frequencies of donor-specific CD4+CD45RA+ T cells. CONCLUSIONS: In renal transplant recipients, donor-specific CD4+ T cell hyporesponsiveness occurs predominantly in CD4+ CD45RO+ T cells which is the subset capable of trafficking through the graft. PMID- 11502980 TI - Comparison of activation requirements and activation phenotype of allogeneic and xenogeneic rodent responses in vivo and in vitro. AB - BACKGROUND: Both discordant and concordant xenogeneic responses are dominated by humoral immunity. Recent advances in molecular engineering approaches may largely prevent rejection by means of this pathway, leaving the cellular arm of the immune response as the principal remaining barrier to successful engraftment. METHODS: To characterize further the cellular response to xenogeneic tissues, we used the intracellular fluorescent marker CFSE (5-(and-6)-carboxyfluorescein diacetate succinimidyl ester) to track the mitotic record of T cells (and T cell subsets) after either xenogeneic or allogeneic activation in vitro or in vivo. Activation marker expression was monitored by simultaneous labeling with antibodies for either CD25 or CD134. RESULTS: The in vitro and in vivo responses of Lewis lymphocytes were generally similar in magnitude and timing comparing activation with allogeneic or xenogeneic stimulators. However, the xenogeneic T cell precursor frequency was found to be markedly higher than that previously reported and were comparable to that seen in allogeneic responses. Xenogeneic responses were unique in the continued expression of activation markers in later division cycles. In addition, CD4 and CD8 T-cell proliferation was highly dependent on stimulator class II expression, highlighting the importance of CD4 T cells and the indirect pathway in the xenogeneic response. CONCLUSIONS: An unexpectedly high precursor frequency was detected for xenogeneic cellular responses in the rat anti-mouse combination and was comparable to that seen in allogeneic responses. Differences in xenogeneic versus allogeneic activation profiles exist that may result from the cellular pathways used for activation. PMID- 11502981 TI - Molecular pathogenesis of Epstein-Barr virus associated posttransplant lymphomas: new insights through latent membrane protein 1 fingerprinting. AB - BACKGROUND: Fingerprint amino acid patterns within the carboxy terminus of the latent membrane protein (LMP1) oncoprotein of Epstein-Barr virus (EBV) allow individual strain identification at the molecular level. LMP1 is expressed in the tumor cells of EBV-associated posttransplant lymphomas (PTLs) and the LMP1 genome is also identified in lymphocytes of most donors of allogeneic bone marrow. Therefore, LMP1 genotyping in donor lymphocytes and PTL tumor cells, together with sex chromatin determination of tumor cells, allows to determine the origin of PTL tumor cells and the origin of individual EBV strains harboured by them. METHODS: We traced the origin of aggressive PTLs occurring in six patients after allogeneic T cell-depleted stem cell transplantation (allo-SCT). DNA was extracted from donor lymphocytes and PTLs of recipients and amplified with LMP1 specific primers in each case. A comparative sequence analysis of the fingerprint LMP1 region identified in donor lymphocytes and lymphoma was performed. RESULTS: One lymphoma of donor origin occurred after highly selected CD34+ PBSCT and contained the same LMP1 genotype as the donor lymphocytes. Three lymphomas of recipient origin had deletions within the carboxy terminus of LMP1, not identified in the donor strains. All lymphomas occurred in the setting of allo SCT and had a rapid clinical course. CONCLUSIONS: These results show that highly selected CD34+ PBSCT does not protect against transfer of EBV positive founder cells of donor type PTL and that, after allo-SCT, recipient type PTLs are not uncommon. Outgrowth of recipient type lymphoma may be favoured by LMP1 deletion variant strains present in recipient lymphocytes. PMID- 11502982 TI - Immunotoxin FN18-CRM9 induces stronger T cell signaling than unconjugated monoclonal antibody FN18. AB - BACKGROUND: The T-cell receptor (TCR)/CD3 complex is the target of therapeutic strategies aimed at prolonging allograft survival. The immunotoxin FN18-CRM9, composed of the anti-CD3 monoclonal antibody FN18 and the mutated diphtheria toxin CRM9, is useful for prolonging allograft survival in preclinical models of transplantation. To explore the influence of conjugation of the mutated diphtheria toxin on functional activation of the TCR/CD3 complex, we compared the effects of FN18-CRM9 and unconjugated FN18 on protein tyrosine phosphorylation and ligand/receptor internalization in purified monkey peripheral blood T cells. METHODS: Purified normal rhesus monkey T cells were incubated with unconjugated FN18 or conjugated FN18-CRM9 and examined for differences in antibody binding, tyrosine phosphorylation, and CD3 internalization. RESULTS: Binding cross inhibition studies demonstrated that both compounds were able to inhibit fluorescein isothiocyanate-FN18 binding to CD3 with similar efficacy and potency. However, FN18-CRM9 was more potent than FN18 in triggering the phosphorylation of several proteins on tyrosine residues and in inducing CD3 internalization. The tyrosine kinase inhibitor genistein blocked FN18-CRM9-induced protein tyrosine phosphorylation and CD3 internalization, suggesting that tyrosine phosphorylation is involved in the internalization of the immunotoxin. Interestingly, in FN18 CRM9- but not FN18-treated cells, there was a gradual decrease in cellular CD3 protein levels within 24 and 48 hr; such a decrease was not observed with the control protein Csk. CONCLUSIONS: Our findings suggest that the conjugation of the mutated diphtheria toxin CRM9 to FN18 modulates the monoclonal antibody mediated cross-linking of the TCR/CD3 complex, leading to a stronger protein tyrosine phosphorylation and CD3 internalization. This may in turn contribute to the greater efficacy of the immunotoxin in prolonging allograft survival. PMID- 11502983 TI - Human anti-porcine gammadelta T-cell xenoreactivity is inhibited by human FasL expression on porcine endothelial cells. AB - BACKGROUND: The role of gammadelta T cells during an immune response is still elusive and has been proposed to play a first line of defense along with other cells of the innate immune system, such as macrophages and natural killer cells, before alphabeta T-cell activation occurs. Innate cellular immune response plays a major role in xenograft rejection. We investigated the response of human gammadelta T cells to unmodified and human FasL (hFasL)-expressing xenogenic porcine endothelial cells. METHODS: A 51Cr release assay was used to study the xenoreactivity of human gammadelta T-cell clones against porcine endothelial cells. Stable transfectants of porcine endothelial cells expressing hFasL were established and analyzed for their effectiveness in controlling this response. RESULTS: Of the gammadelta T-cell clones tested, 38.9% were cytotoxic for porcine endothelial target cells. This cytotoxic response of human gammadelta T-cell clones was significantly inhibited by a monoclonal antibody against human CD3. Incubation of gammadelta T-cell clones with concanamycin A, an inhibitor of the perforin/granzyme B pathway, caused inhibition of lysis of porcine endothelial cells. Inhibition was not observed upon incubation with either anti-FasL or anti tumor necrosis factor-alpha monoclonal antibodies. Expression of hFasL on porcine endothelial cells significantly reduced lysis by human gammadelta T cells. CONCLUSION: These results imply that human gammadelta T cells may represent an important obstacle to xenotransplantation. Specific strategies targeted at this subset of T cells could be important in controlling innate cellular response to xenografts and facilitate graft survival. PMID- 11502984 TI - Platelet-mediated activation of endothelial cells: implications for the pathogenesis of transplant rejection. AB - BACKGROUND: Platelets exert their normal functions at sites of endothelial disruption by plugging discontinuities in blood vessels and secreting products that promote thrombosis, inflammation, and the healing of wounds. Whether platelets might induce these changes in xenograft blood vessels, leading to development of acute vascular rejection, has been uncertain. METHODS: To examine the role of human platelets in modulation of xenograft endothelium, pig endothelial cells were treated with human platelets. RESULTS: Treatment of quiescent porcine endothelial cells with human platelets modulated the endothelial cells. Whereas resting human platelets caused little change in normal porcine endothelial cells, platelets activated with small amounts of thrombin induced striking changes in the endothelial cells, including the induction of tissue factor activity, the expression of E-selectin, and the secretion of endothelin-1. These changes were induced, at least in part, by interleukin-1 (IL 1) associated with the platelet surface and were modified by the secretion of transforming growth factor-beta (TGF-beta). CONCLUSION: These findings may explain how the activation of platelets at an early point in the rejection of vascularized organ xenografts or in chronic diseases might contribute to thrombotic, ischemic, and inflammatory changes characteristic of an organ xenograft undergoing rejection. PMID- 11502985 TI - Biphasic response of the regional lymphatics in the normal lymphocyte transfer reaction. AB - BACKGROUND: Initially developed for histocompatibility testing, the normal lymphocyte transfer (NLT) reaction involves the intradermal injection of allogeneic lymphocytes from one individual to another. Because of the unique kinetics of the immunological response to allogeneic lymphocytes, the NLT reaction has been considered an informative system for the analysis of transplant immunity. METHODS: In this study, we used bilateral efferent lymph duct cannulations in sheep to examine the regional lymphatic response to the NLT reaction. Our studies used monoclonal antibodies to define lymphocyte population dynamics and DNA flow cytometry to reflect lymphocyte proliferative responses. RESULTS: The results confirmed a biphasic NLT reaction. An unexpected finding was the marked differences between the early and late NLT responses. The early response was characterized by T-lymphocyte proliferation, as reflected by S-phase DNA, which was comparable in both the NLT-stimulated and contralateral control efferent lymphocytes. This bilateral proliferative response was observed in both CD4+ and CD8+ lymphocytes. In contrast, the late response was restricted to the efferent lymph from the NLT-stimulated lymph node. Dual-parameter flow cytometry demonstrated that the dominant component of this unilateral NLT response was CD8+ lymphocytes. CONCLUSIONS: These results suggest important functional distinctions between systemic and regional lymphatic responses to intradermal alloantigens. PMID- 11502986 TI - Peripheral regulation of graft-versus-host alloreactivity in mixed chimeric miniature swine. AB - BACKGROUND: Despite the presence of circulating donor-derived T cells during the induction of mixed chimerism across MHC barriers in miniature swine, severe graft versus-host disease was avoided in the majority of animals. In this study, we investigated the possible roles of recipient and donor lymphoid populations in the regulation of donor-anti-recipient alloreactivity. METHODS: Mixed chimerism across a full MHC-mismatch barrier was established in miniature swine using a high-dose allogeneic peripheral blood stem cell protocol. Peripheral blood mononuclear cells from mixed chimeric swine were co-cultured with naive donor matched responders and naive recipient-matched stimulators in mixed lymphocyte reactions. RESULTS: Peripheral blood mononuclear cells from mixed chimeras inhibited donor-anti-recipient proliferation. This suppression was radioresistant to 25 Gy. Suppression of donor-anti-recipient alloreactivity was not observed in mixed lymphocyte co-cultures when donor-derived cells were added in the absence of recipient-derived cells. CONCLUSIONS: These results suggest an association between the presence of an active and relatively radioresistant cell population, demonstrable in vitro, and the regulation of graft-versus-host disease across MHC barriers in mixed chimeric miniature swine. PMID- 11502987 TI - Evans' syndrome complicating chronic graft versus host disease after cadaveric liver transplantation. AB - Acute graft versus host disease (GVHD) occurred in a patient after cadaveric liver transplantation from an HLA disparate donor. Immunosuppression resulted in a remission, but chronic GVHD with a scleroderma-like syndrome ensued. This was further complicated by immune hemolytic anemia and thrombocytopenia (Evan's syndrome). Semi-quantitative microsatellite analysis of circulating lymphoid cells showed that T cells were predominantly of donor origin, thereby explaining the chronic GVHD. The marrow hematopoietic cells remained recipient, so that the immune cytopenias were expected to be alloimmune in nature. However, the red cell antibodies were shown to have anti-C and anti-e specificity, with both the donor (R1R1) and recipient (R1r) possessing the C and e antigens. Therefore, the immune hemolysis might be considered both alloimmune and autoimmune. The patient finally died of sepsis. This case illustrates that chronic GVHD due to stable donor T cell engraftment may rarely occur in liver transplantation despite HLA disparity. Immunosuppression may result in dysregulation of T cell functions, leading to alloimmune and autoimmune problems. PMID- 11502988 TI - Successful use of cardiac allograft from serotonin antagonist intoxication. AB - BACKGROUND: Compromised organ donors are generally not accepted for heart transplantation (HT) despite the increasing number of critically ill patients on the waiting lists. By extending the donor criteria to include certain cases of intoxication, the organ shortage may be reduced. METHODS: The case of a successful orthotopic HT with an allograft from a donor poisoned by antidepressant overdose is presented. RESULTS: Early graft function was satisfactory with anteroseptal dyskinesis and an ejection fraction of 75% on echocardiography. The cardiac allograft recipient suffered some postoperative complications including gastrointestinal problems. The following period was up to now uneventful. Discharge from the intensive care unit was after 4 days. In hospital stay was prolonged at 26 days. CONCLUSIONS: Because of limited myocardial toxicity, donor hearts from certain victims of antidepressant intoxication may be safely used for HT. Existing cardiac organ donor criteria must be reevaluated to maximise the available organ pool. PMID- 11502989 TI - Diagnostic yield of screening colonoscopies in lung transplant candidates. AB - BACKGROUND: Colonoscopy has been used to screen lung transplant candidates for colorectal diseases that would preclude transplantation. The diagnostic yield of this procedure is unknown. METHODS: This is a retrospective cohort study of patients 50 years of age and over who underwent lung transplant evaluations from 1996 to 1999. We assessed the prevalence and location of colonoscopic abnormalities, the predictive value of risk factors for colonic neoplasms, and the impact of colonoscopic findings on management. RESULTS: Thirty-one patients were evaluated. Twenty-four patients had at least one abnormal endoscopic finding. Six patients (19%) had adenomatous polyps; no carcinomas were detected. The 13 patients with risk factors were more likely to have adenomas (relative risk=2.8, P=0.2). The negative predictive value of risk factors for adenomas was 89%. One patient's management was altered and none were denied transplant listing because of the colonoscopic findings. CONCLUSIONS: Screening colonoscopy did not substantively alter the management of lung transplant candidates. More selective screening strategies may be warranted. PMID- 11502990 TI - Successful renal transplantation after endovascular bifurcated stent graft repair of an abdominal aortic aneurysm. AB - Renal transplantation after repair of aortoiliac aneurysms with traditional prosthetic vascular grafts has been shown to be effective. Vascular surgery continues to rapidly evolve, most notably with the advancement of endovascular repair of abdominal aortic aneurysms. Controlled trials continue to support the trend toward the use of endovascular bifurcated aortic stent grafts. For this we describe the first renal transplant in a patient with an endovascular bifurcated aortoiliac stent graft. No intraoperative difficulties were encountered. At 1 year follow-up, the transplanted kidney is functioning well with a normal serum creatinine level of 1.3 mg/dl, and the patient has no worsening of peripheral vascular disease. We recommend that the presence of an endovascular aortic graft not be a contraindication to renal transplantation. PMID- 11502991 TI - Nosocomial outbreak of influenza virus A (H3N2) infection in a solid organ transplant department. AB - BACKGROUND: There is a strong body of evidence in favor of influenza virus immunization in solid organ recipients. However, little attention has been devoted to other reservoirs, such as the patients' relatives and, at the time of hospital admission, to the healthcare workers. METHODS: Analysis of the epidemiology of an outbreak of nosocomial influenza A in a solid organ transplant unit. RESULTS: Four cases of influenza A virus infection were reported during a short 4-day outbreak in a 12 single-room transplant unit. None of the patients had been immunized against influenza. Three patients had not been visited by their relatives between admission and influenza infection. Three nurses, among the 27 healthcare workers, presented with clinical flu symptoms at times consistent with nosocomial transmission. CONCLUSIONS: Because the prevention of influenza infection by vaccination warrants a global strategy to target the different reservoirs, we suggest that the modern policy of vaccinating solid organ patients should be extended both to their relatives and to the healthcare workers of transplant units. PMID- 11502992 TI - A giant multiacinar macroregenerative nodule in an explanted liver. AB - Macroregenerative nodules (MRN) have been detected with increased incidence in explanted livers since orthotopic liver transplantation (OLTx) has become a routine treatment for end-stage liver disease. Autopsy series suggest that MRN may be more common than once thought, and several studies point to the malignant potential of these lesions. With increasing waiting times for OLTx, the propensity for these premalignant lesions to arise in cirrhotic livers has important implications for the supervision of patients on OLTx waiting lists. We present here a striking example of a MRN and review a topic that is generating considerable interest. PMID- 11502993 TI - Fate of a renal tubulopapillary adenoma transmitted by an organ donor. AB - Organ transplantation from cadaveric donors has a risk of cancer transmission. However, some reports indicate that kidneys bearing small carcinomas can be safely transplanted, as can other organs harvested from the same donor. We report herein the case of two allograft recipients (left kidney and heart with no evidence of tumor) who developed a renal carcinoma soon after transplantation. The initial tumor of the donor was a 17-mm tubulopapillary adenoma found on the right kidney, which was not transplanted. The left kidney recipient rejected all residual tumoral cells after graft removal and immunosuppression discontinuation. The heart recipient died 7 months after transplantation from metastasis of a renal carcinoma. This strongly suggests that circulating carcinoma cells were present at the time of organ retrieval and that they were not cleared by in situ perfusion. In contrast with the literature data, this report indicates that patients with small renal tubulopapillary tumors should not be considered for organ donation. PMID- 11502994 TI - Comparison of the effects of enalapril and losartan on posttransplantation erythrocytosis in renal transplant recipients: prospective randomized study. AB - BACKGROUND: The aim of this prospective randomized study was to compare the safety and efficacy of enalapril (E) and losartan (L) in the treatment of posttransplantation erythrocytosis and the effect of the ACE genotype on response to therapy. METHODS: Twenty-seven (24 male and 3 female, mean age 34+/-8 years) renal transplant recipients with erythrocytosis were treated either with E (15 patients) (10 mg/day) or L (12 patients) (50 mg/day) for 8 weeks. RESULTS: The hemoglobin levels were significantly decreased in the L (17.1+/-0.7 to 15.9+/-1.3 g/dl, P=0.01) and E groups (17.4+/-1.1 to 14.9+/-2.2 g/dl, P=0.001). Among the responders who discontinued treatment, there was a trend for longer time to relapse in the L group (7.38+/-3.75 months; 95% confidence interval: 0.03-14.7) compared with the E group (2.75+/-0.70 (95% confidence interval: 1.37-4.13) (P=0.11). Decrease in hemoglobin was more prominent with E compared with L ( 3.26+/-0.65 vs. -1.70+/-0.39 g/dl, P=0.05). Decrease in hemoglobin levels between DD and non-DD genotype groups was similar (-2.0+/-1.5 vs. -1.7+/-2.3 g/dl, P=0.69). CONCLUSIONS: Enalapril caused a greater decrease but faster relapse in hemoglobin levels compared with losartan in patients with posttransplantation erythrocytosis. The DD type polymorphism had no effect on response. PMID- 11502995 TI - Safety of aerosolized amphotericin B lipid complex in lung transplant recipients. AB - BACKGROUND: Fungal infections remain an important cause of morbidity and mortality in lung transplant recipients. Aerosolized amphotericin B lipid complex (ABLC) may be more efficacious than conventional amphotericin B in the prevention of fungal infections in animal models, but experience with aerosolized ABLC in humans is lacking. METHODS: We conducted a prospective, noncomparative study designed to evaluate safety of aerosolized ABLC in lung or heart-lung transplant recipients. RESULTS: A total of 381 treatments were administered to 51 patients. Complete spirometry records were available for 335 treatments (69 in intubated patients, 266 in extubated patients). ABLC was subjectively well tolerated in 98% of patients. Pulmonary mechanics worsened by 20% or more posttreatment in less than 5% of all treatments. There were no significant adverse events related to study medication in any patient, and 1-year survival for all enrolled patients was 78%. CONCLUSION: Administration of nebulized ABLC is safe in the short-term and well-tolerated in lung transplant recipients. Additional prospective, randomized studies are needed to determine the efficacy of aerosolized ABLC alone or in conjunction with systemic therapies in the prevention of fungal infections in lung transplant recipients. PMID- 11502996 TI - Heterozygosity for the factor V Leiden (G1691A) mutation predisposes renal transplant recipients to thrombotic complications and graft loss. AB - BACKGROUND: Heterozygosity for a mutation in the coagulation factor V gene (factor V Leiden; FVL) leads to resistance to activated protein C and represents the most common cause of inherited thrombophilia. FVL is associated with a high risk for thromboembolic events and might be a risk factor for venous thrombosis and early graft loss in renal transplant recipients. METHODS: We studied a cohort of 202 renal allograft recipients to assess the impact of the FVL mutation on thrombotic events and graft loss within 1 year after transplantation. We recorded the occurrence of deep venous thrombosis, pulmonary embolism, early graft perfusion defect, and graft loss. The occurrence of these events was then correlated with the presence or absence of heterozygosity for the FVL mutation. RESULTS: Heterozygosity for FVL was detected in 8 (4%) of 202 patients. The incidence of deep venous thrombosis or pulmonary embolism was higher in heterozygous compared with wild-type patients (25% vs. 5.7%, P=0.09). Furthermore, early graft perfusion defect (25% vs. 2.6%; P=0.03) and graft loss within 7 days after transplantation (2/8 vs. 1/194; P=0.004) were significantly more frequent among heterozygous carriers of FVL. All eight FVL carriers were negative for protein C or S deficiency and antiphospholipid and anticardiolipin antibodies, and were not carriers of the G20210A prothrombin mutation. CONCLUSIONS: Heterozygosity for the FVL mutation predisposes renal allograft recipients to venous thromboembolic complications, graft perfusion defects, and early transplant loss. Screening for the FVL mutation and appropriate peri- and postoperative anticoagulation after renal transplantation might prevent these thromboembolic complications. PMID- 11502997 TI - Cerivastatin-induced rhabdomyolysis in a renal transplant on cyclosporin. PMID- 11502998 TI - Living donor with type IV-A choledochal cyst in liver transplantation. PMID- 11502999 TI - Homologous transplantation of prepubertal testes in the dog. PMID- 11503000 TI - Neutropenia after treatment of posttransplantation erythrocytosis with enalapril. PMID- 11503001 TI - Expanded criteria donors/high risk recipients is not always cheaper than hemodialysis. PMID- 11503002 TI - Programmed cell death in extraocular muscle tendon/sclera precursors. AB - PURPOSE: This study was designed to examine the occurrence of natural cell death in the periocular mesenchyme of mouse embryos. METHODS: Vital staining with LysoTracker Red and Nile blue sulfate as well as terminal nick end labeling (TUNEL) were utilized to identify apoptotic cell death in whole and histologicaly sectioned gestational day 10.5 to 14 mouse embryos. Laser scanning confocal microscopy was used to provide a three dimensional representation of the cell death pattern. Immunohistochemical staining for neural crest and myoblast populations was utilized to indicate the cell population undergoing apoptosis. RESULTS: Programmed cell death was evident in the developing rectus muscle tendons/sclera on gestational days 11 through 12.5 (corresponding to the weeks 5 6 of human development). Although each of these peripheral periocular condensations has readily apparent amounts of apoptosis, the pattern of cell death varied among them. Cell death was most apparent in the superior rectus tendon primordium, while that for the lateral rectus had the least evidence of apoptosis. CONCLUSIONS: Although apoptosis was readily evident in the periocular mesenchyme in distinct regions located medial and distal to the developing rectus muscles, programmed cell death in these sites has not previously been reported. New imaging techniques coupled with stains that evidence apoptotic cell death have made it possible to define this tissue as a prominent region of programmed cell death. Although neuronal tissues, including particular regions of the developing eye, are well recognized as sites of programmed cell death, description of this phenomenon in the extraocular tendon/sclera precursors is novel. PMID- 11503003 TI - Optimizing drug development: strategies to assess drug metabolism/transporter interaction potential-toward a consensus. PMID- 11503004 TI - Demonstrating effectiveness in a post-placebo era. PMID- 11503005 TI - The effect of pregnancy on cytochrome P4501A2, xanthine oxidase, and N acetyltransferase activities in humans. AB - OBJECTIVE: Our objective was to evaluate the activity of cytochrome P4501A2 (CYP1A2), xanthine oxidase (XO), and N-acetyltransferase 2 (NAT2) from early to late pregnancy and after delivery. METHODS: Twelve women were studied on three occasions during pregnancy (early, 8-16 weeks' gestation; middle, 20-28 weeks' gestation; and late, 32-39 weeks' gestation) and about 1 month after delivery. Caffeine was used as a metabolic probe. After the women ingested a can or a bottle of caffeine-containing soft drink, urine samples were collected for 12 hours. The caffeine metabolites measured were 5-acetylamino-6-amino-3 methyluracil (AAMU), 1-methylxanthine (1X), 1-methyl-uric acid (1U), 1,7-dimethyl uric acid (17U), and 1,7-dimethylxanthine (17X). The hepatic enzyme activities were estimated by the urinary caffeine metabolic ratios as follows: CYP1A2 = (AAMU + 1X + 1U)/17U; XO = 1U/(1X + 1U); NAT2 = AAMU/(AAMU + 1X + 1U). RESULTS: Statistically significant differences were found in CYP1A2 (P < .0001) and NAT2 (P < .01). The mean metabolic ratios for CYP1A2 during pregnancy (6.80, 5.18, and 4.97 for the early phase, middle phase, and late phase, respectively) were significantly lower than the ratio after delivery (10.39). The mean metabolic ratio for NAT2 in the early phase (0.57) was significantly lower than after delivery (0.66). There was no significant difference in metabolic ratios for XO during pregnancy and after delivery. CONCLUSION: The data demonstrate that pregnancy influences CYP1A2 and NAT2 activity. CYP1A2 activity decreases not only in late pregnancy but also in early and middle pregnancy. PMID- 11503006 TI - Effect of ranitidine on the pharmacokinetics of triazolam and alpha hydroxytriazolam in both young (19-60 years) and older (61-78 years) people. AB - OBJECTIVE: This study evaluated the effect of oral ranitidine (75 mg and 150 mg) on the pharmacokinetics of triazolam (0.25 mg) and its major metabolite, alpha hydroxytriazolam, in both young and older people. Metabolite data were used to distinguish the mechanism of this interaction. METHOD: This was a randomized, open-label, 3-way crossover study. Eighteen young (19-60 years) and 12 older (61 78 years) men and women were randomly assigned to receive evening doses of triazolam 0.25 mg (1) alone, (2) on the third day of dosing ranitidine 75 mg twice daily for 4 days, and (3) on the third day of dosing ranitidine 150 mg twice daily for 4 days. RESULTS: In the young group, mean triazolam area under the concentration-time curve from time zero to infinity [AUC(0-infinity)] was 10% and 28% higher after treatment with 75 mg and 150 mg ranitidine, respectively. In the older group, mean triazolam AUC(0-infinity) was 31% and 28% higher after treatment with 75 mg and 150 mg ranitidine, respectively. There was no change in the alpha-hydroxytriazolam/triazolam AUC(0-infinity) ratio in either age group, indicating that neither formation nor elimination of alpha-hydroxytriazolam was affected by ranitidine. There were no changes in the half-life of triazolam or alpha-hydroxytriazolam. CONCLUSION: Ranitidine increases oral absorption of triazolam in both young and older people. This effect is likely caused by elevation of gastrointestinal pH, allowing for greater absorption of acid-labile triazolam. The difference in this effect between age groups at the lower 75-mg dose of ranitidine suggests that older people may be more sensitive to the antisecretory effect of ranitidine. PMID- 11503007 TI - Modulation of the cytochrome P450-mediated metabolism of ifosfamide by ketoconazole and rifampin. AB - BACKGROUND: The autoinducible metabolic transformation of the anticancer agent ifosfamide involves activation through 4-hydroxyifosfamide to the ultimate cytotoxic ifosforamide mustard and deactivation to 2- and 3 dechloroethylifosfamide with concomitant release of the neurotoxic chloroacetaldehyde. Activation is mediated by cytochrome P450 (CYP) 3A4 and deactivation by CYP3A4 and CYP2B6. The aim of this study was to investigate modulation of the CYP-mediated metabolism of ifosfamide with ketoconazole, a potent inhibitor of CYP3A4, and rifampin (INN, rifampicin), an inducer of CYP3A4/CYP2B6. METHODS: In a double-randomized, 2-way crossover study a total of 16 patients received ifosfamide 3 g/m(2) per 24 hours intravenously, either alone or in combination with 200 mg ketoconazole twice daily (1 day before treatment and 3 days of concomitant administration) or 300 mg rifampin twice daily (3 days before treatment and 3 days of concomitant administration). Plasma pharmacokinetics and urinary excretion of ifosfamide, 2- and 3 dechloroethylifosfamide, and 4-hydroxyifosfamide were assessed in both courses. Data analysis was performed with a population pharmacokinetic model with a description of autoinduction of ifosfamide. RESULTS: Rifampin increased the clearance of ifosfamide at the start of therapy at 102%. The fraction of ifosfamide metabolized to the dechloroethylated metabolites was increased, whereas exposure to the metabolites was decreased as a result of increased elimination. The fraction metabolized and the exposure to 4-hydroxyifosfamide were not significantly influenced. Ketoconazole did not affect the fraction metabolized or the exposure to the dechloroethylated metabolites, whereas both parameters were reduced with 4-hydroxyifosfamide. CONCLUSIONS: Coadministration of ifosfamide with ketoconazole or rifampin did not produce changes in the pharmacokinetics of the parent or metabolites that may result in an increased benefit of ifosfamide therapy. PMID- 11503008 TI - Erythromycin increases plasma concentrations of alpha-dihydroergocryptine in humans. AB - OBJECTIVE: Our objective was to investigate the potential for relevant pharmacotherapeutic interaction between cytochrome P4503A4 (CYP3A4)-inhibiting agents such as erythromycin and the dopamine agonist alpha-dihydroergocryptine (DHEC). METHODS: The study was carried out as a single-center, controlled, nonblinded, 2-way crossover clinical trial with randomly allocated period balanced sequences, investigating two treatments of a single oral dose of 10 mg DHEC (on the morning of day 1), once administered alone (reference), once along with a 4-day treatment (days -2 to 1) of 500 mg erythromycin 3 times daily. Periods were separated by a washout of at least 14 days. Nine healthy white male volunteers, 22 to 42 years old, with a body weight range of 58 to 90 kg (body mass index, 20.2-25.1 kg x m(-2)) began the study. One subject discontinued prematurely, and 8 concluded the study in accordance with the study protocol. RESULTS: The plasma and urinary pharmacokinetics of DHEC and its metabolites were characterized by a large variability. Concomitant treatment with erythromycin led to respective increases of 9.5 (95% confidence interval [CI], 6.5 to 13.9) and 16.5 (95% CI, 8.7 to 31.5) times the maximum observed plasma drug concentration and the area under the time course of the plasma concentrations up to the last quantifiable concentration after dosing of unchanged DHEC (determined by radioimmunoassay). The 24-hour urinary excretion was on average 11 times larger (95% CI, 5.9 to 20.7). Qualitatively similar findings were recorded for the total of DHEC plus metabolites (as determined by enzyme immunoassay). CONCLUSIONS: The concomitant use of erythromycin or similarly CYP3A4-inhibiting agents along with direct dopaminergic agonists such as the ergoline DHEC may cause a clinically relevant increase in pharmacokinetic exposure, which may induce exaggerated dopaminergic effects. PMID- 11503010 TI - Interindividual variability in sensitivity to warfarin--Nature or nurture? AB - BACKGROUND: Interindividual variability in responses to warfarin is attributed to dietary vitamin K, drug interactions, age, or genetic polymorphism in the cytochrome P4502C9 enzyme (CYP2C9) (allelic variants 2C9*2 and 2C9*3 ) linked with impaired metabolism of the potent enantiomere S-warfarin. PATIENTS AND METHODS: We quantified the relative effects of age and of simultaneously determined CYP2C9 genotype, plasma warfarin and vitamin K concentrations, and concurrent medications on warfarin maintenance doses in 156 patients at optimized stable anticoagulation. RESULTS: Allele frequencies for CYP2C9*1, CYP2C9*2, and CYP2C9*3 were 0.84, 0.10, and 0.06. Warfarin doses were 6.5 +/- 3.2, 5.2 +/- 2.4, and 3.3 +/- 2.0 mg/d in the 3 genotype groups (P < .0001). Warfarin doses decreased with age as follows: 7.7 +/- 3.7 versus 4.9 +/- 2.9 mg/d at < 50 years and >66 years (P < .001), mainly as a result of decreased plasma warfarin clearance (2.8 +/- 1.4 mL/min versus 1.9 +/- 0.8 mL/min; P < .001). Vitamin K (1.6 +/- 1.1 ng/mL) did not differ among the age or genotype groups. Patients >or=66 years old with the CYP2C9*3 allele required only 2.2 +/- 1.2 mg/d compared with 7.9 +/- 3.7 mg/d in those C transition at position -1912 (T-1912C), C-1886G, C-1566T, G-1538A, C-1189T, G-982A, and A-162G were 0.019, 0.019, 0.077, 0.019, 0.579, 0.019, and 0.003, respectively. Some mutations occurred simultaneously, and a total of 6 sequence patterns (patterns 1 6) were observed. The luciferase reporter gene assay indicated that the presence of mutation(s) resulted in a reduction in luciferase activity of 41.4% (pattern 2) to 86.8% (pattern 5) compared with the activity of the wild-type construct. The calculated intrinsic clearance of phenytoin was also lower (up to a 40% reduction for pattern 2) when a mutation(s) was present. CONCLUSION: In addition to the two major mutations in the coding region (CYP2C9*2 and CYP2C9*3 ), mutations in the 5'-flanking region of the human CYP2C9 gene appear to contribute to the large interindividual variability in drug metabolism activity. PMID- 11503009 TI - A pharmacokinetic study of intermittent rifabutin dosing with a combination of ritonavir and saquinavir in patients infected with human immunodeficiency virus. AB - AIM: Our primary aim was to evaluate the plasma exposures and safety of rifabutin and its active 25-O-desacetyl metabolite during concomitant therapy of intermittent rifabutin dosing regimens with a combination of ritonavir and saquinavir. METHODS: Twenty-four patients without mycobacterial infection who were human immunodeficiency virus seropositive and who were receiving 400 mg each of ritonavir and saquinavir twice daily participated in a 3-period, 2-group longitudinal pharmacokinetic study. Patients were equally randomized to receive 300 mg of rifabutin every 7 days (group 1) or 150 mg of rifabutin every 3 days (group 2) for 8 weeks. Blood samples were collected over the dosing intervals of the protease inhibitors at baseline (period 1) and of the 3 drugs after 4 weeks (period 2) and 8 weeks (period 3) for HPLC measurement of plasma concentrations of the 3 drugs and 25-O-desacetylrifabutin. RESULTS: Nineteen patients (group 1, n = 10; group 2, n = 9) completed the study. Five individuals withdrew from the study; 3 of them experienced side effects, and 2 were lost to follow-up. For combined groups, mean saquinavir and ritonavir overall (area under the concentration-time curve [AUC]) and peak (C(max)) plasma exposures averaged over periods 2 and 3 did not change significantly (8% to 19%; P > .05) compared with those in period 1 (90% confidence intervals, -7% to 26% for ritonavir and -2% to 38% for saquinavir). Rifabutin and metabolite AUC and C(max) exposures were stable over the 8 weeks, with intraindividual coefficients of variation of 12% to 19%. Oral clearance of rifabutin was similar in both groups (321 mL/min in group 2 versus 372 mL/min in group 1; P = .34). Rifabutin C(max) values were significantly lower in group 2 (310 ng/mL versus 496 ng/mL in group 1; P = .004). Rifabutin and metabolite predose levels were significantly higher in group 2 (rifabutin: 54 ng/mL versus 17 ng/mL; desacetyl rifabutin: 55 ng/mL versus 28 ng/mL; P < .002). CONCLUSIONS: Rifabutin exposures were similar at 4 and 8 weeks and had minimal effect on ritonavir and saquinavir exposures. Intermittent rifabutin dosing over 8 weeks provided a safe and manageable regimen for concurrent therapy with a combination of ritonavir and saquinavir. PMID- 11503013 TI - Detection of one single mutation predicts thiopurine S-methyltransferase activity in a population of Saami in northern Norway. AB - Thiopurine S-methyltransferase (TPMT) activity exhibits genetic polymorphism. The purpose of this investigation was to identify TPMT mutant alleles in the Saami population as a basis of developing genotyping tests for prediction of TPMT activity. The most predominant allele in Saamis (n = 194) was the TPMT*3C allele (A719G mutation) representing 92% of the mutant alleles, with an estimated allelic frequency of 3.3%. The most frequent allele in Caucasians (n = 66) living in the same geographic area was the TPMT*3A (A719G and G460A mutations) representing 91% of the mutant alleles, with an estimated allelic frequency of 3.4%. A test for one mutation, A719G, may prospectively identify more than 90% of the Saami individuals who require reduction in thiopurine dose to avoid hematopoietic toxicity. In a Norwegian population, comprising both the major Caucasian population and a minor Saami population, the same genotyping tests (eg, tests for the A719G and G460A mutations) may be used. PMID- 11503014 TI - Identification of functionally variant MDR1 alleles among European Americans and African Americans. AB - MDR1 (P-glycoprotein) is an important factor in the disposition of many drugs, and the involved processes often exhibit considerable interindividual variability that may be genetically determined. Single-strand conformational polymorphism analysis and direct sequencing of exonic MDR1 deoxyribonucleic acid from 37 healthy European American and 23 healthy African American subjects identified 10 single nucleotide polymorphisms (SNPs), including 6 nonsynonymous variants, occurring in various allelic combinations. Population frequencies of the 15 identified alleles varied according to racial background. Two synonymous SNPs (C1236T in exon 12 and C3435T in exon 26) and a nonsynonymous SNP (G2677T, Ala893Ser) in exon 21 were found to be linked (MDR1*2 ) and occurred in 62% of European Americans and 13% of African Americans. In vitro expression of MDR1 encoding Ala893 (MDR1*1 ) or a site-directed Ser893 mutation (MDR1*2 ) indicated enhanced efflux of digoxin by cells expressing the MDR1-Ser893 variant. In vivo functional relevance of this SNP was assessed with the known P-glycoprotein drug substrate fexofenadine as a probe of the transporter's activity. In humans, MDR1*1 and MDR1*2 variants were associated with differences in fexofenadine levels, consistent with the in vitro data, with the area under the plasma level time curve being almost 40% greater in the *1/*1 genotype compared with the *2/*2 and the *1/*2 heterozygotes having an intermediate value, suggesting enhanced in vivo P-glycoprotein activity among subjects with the MDR1*2 allele. Thus allelic variation in MDR1 is more common than previously recognized and involves multiple SNPs whose allelic frequencies vary between populations, and some of these SNPs are associated with altered P-glycoprotein function. PMID- 11503015 TI - [Presence of genetic rearrangements in E1/E2 regions of episomal HPV 16 isolates from cervical carcinomas 5-8]. AB - OBJECTIVE: Analysis of genetic situations of E1/E2 region of episomal HPV 16 isolates from cervical carcinomas. METHODS: Physical statues of HPV 16 DNAs from cervical carcinomas were analyzed by Southern blot hybridization. E1/E2 regions of episomal RPV 16 isolates were amplified by PCR, and the PCR products were cloned and sequenced. RESULTS: Eight isolates from totally 28 cases showed extrachromosomal HPV 16 DNA. PCR cloned sequence analysis revealed that all the eight isolates contained intact E1/E2 regions, whereas 4 isolates had different ratios of genetic rearrangements, including insertion of E1 repeat sequence and several deletions. Only one case showed the same G deletion at position 1138 as HPV 16 prototype. CONCLUSIONS: DNA rearrangements were present in E1/E2 regions of episomal HPV 16 isolates from cervical cancers and might play a role in tumorigenesis. Different viral genotypes in an identical tumor tissue might indicate that the tumor occurring at the same position of a patient may derive from two or even more cell clones. PMID- 11503016 TI - [Studies on the basis of molecular biology of the phase change of influenza A(H1N1) viruses]. AB - OBJECTIVE: To reveal the basis of molecular biology of the phase change of influenza A (H1N1) viruses. METHODS: Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method, using synthetic oligodeoxynucleotide primers. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (Version 1.03) and Editseq (Version 3.69) software. RESULTS: No special amino acid on HA1 protein molecule determining the influenza A(H1N1) virus with O or D phase feature was found. However, there are differences of amino acid sequences at -2, 7, 130 and 139 positions between H1N1 viruses isolated in or before end after 1995. It is very interesting that since 1995, two different genetic lineages of influenza A (H1N1) virus HA gene have been cocirculating in men in China. One lineage is no any deletion of amino acid on HA1 protein molecule. The other one has a deletion at 130 position. Their amino acid sequences were compared with those of H1N1 viruses isolated before 1995. CONCLUSIONS: There is no special amino acid on HA1 domain protein molecule determining the influenza A (H1N1) virus with O or D phase feature was found. Since 1995, two distinguished genetic lineages of influenza A H1N1 virus HA gene have been cocirculating in men in China. PMID- 11503017 TI - [Identification and visualization of virus-like particles in peripheral blood mononuclear cells]. AB - OBJECTIVE: PBMCs from patients with chronic hepatitis C were examined by electron microscopy (EM), immune EM (IEM) combined with immunohistochemistry to trace the infection and morphology of HCV in the PBMCs. METHODS: The PBMCs from 28 patients with chronic hepatitis C were analyzed firstly for HCV RNA and HCV antigens by RT PCR and immunohistochemistry respectively. The PBMCs with positive HCV RNA and HCV antigens were then observed by electron microscopy. RESULTS: The positive rate of HCV RNA and HCV antigens were 77.27% (17/22) and 75.00% (21/28), respectively. Two types of polymorphic HCV-like particles with diameter of approximately 65 nm and 110 nm were observed in cytoplasmic vesicles of the PBMCs from HCV Ag positive patients. 10 patients with HCV Ag over expression were studied by electron microscopy and immune election microscopy. The budding phenomenon of the particles could be also visualized in the cytoplasmic vesicles of the PBMCs. Some ultrastructural changes showed an increased ratio of cytoplasm to nucleus and pyknosis of nucleus was also suggestive of the virus infection. CONCLUSIONS: The EM study demonstrated the infection and replication of HCV in the PBMCs of chronic hepatitis C patients by morphology and morphogenesis. PMID- 11503018 TI - [Studies on clinical trials and immune effect of a new purified rabies vaccine prepared from Vero cells]. AB - OBJECTIVE: To observe the safety and neutralizing antibody response of a new purified rabies vaccine prepared from Vero cells. METHODS: The vaccine used was manufactured in Hainan Institute of Biologicals and licensed by the Ministry of Health for clinical trials. The vaccine was administrated in 30 and 318 volunteers during phase I and II clinical trials with preexposure (day 0, 7, and 28) or post exposure (day 0,3,7,14 and 28) vaccination schedules. The "Verorab (PVRV)", a rabies vaccine made in France was used as control. RESULTS: The results showed that the new purified vaccine did not cause any severe and moderate side reactions, the slight side reaction rate was 12% including those complained by vaccines themselves. The immunological effect was determined by detecting neutralizing antibody with mouse neutralization test (NIH). The positive seroconversion rates of neutralizing antibody were 100% (n= 27, GMT 2.89 IU/ml) and 100% (n= 30, GMT 5.29 IU/ml) respectively by pre- exposure and post exposure schedules. No significant difference was found when compared with France "Verorab" vaccine by preexposure schedule (n= 24, GMT 2.13 IU/ml). CONCLUSIONS: The clinical trials indicated that the purified rabies vaccine prepared from Vero cells only showed slight side reaction and had good neutralizing antibody response. Therefore this new vaccine is safe and protective for the prevention of rabies in human. PMID- 11503019 TI - [Sequence analysis of the 5' and 3' terminal regions of dengue type 2 virus 04 strain]. AB - OBJECTIVE: To analyze the sequences of 5' and 3' terminal region of dengue type 2 virus 04 strain. METHODS: Total RNA was isolated from dengue type 2 virus 04 strain (D2-04) infected C6/36 cells. With this RNA as template, the cDNA of both 5' and 3' termini of D2-04 were amplified using RACE method respectively. The cDNAs were separately inserted into pGEM-T vector and the recombinant plasmids containing the 5' terminus 535 bp and 3' terminus 503 bp were obtained. The nucleotide sequences of the inserted cDNA fragment were determined. The nucleotide sequences of the 5' and 3' noncoding regions of D2-04 were compared with other dengue type 2 viruses such as JAM, NGC, S1, 16681 and PD-53. RESULTS: The results showed that they shared 98.96%, 98.96%, 93.75%, 98.95%, 97.92% and 97.72%, 97.80%, 90.65%, 94.26%, 94.22% homology with D2-04 strain respectively. CONCLUSIONS: Except SI, the homology between D2-04 and other type 2 viruses was higher than 94%. PMID- 11503020 TI - [Isolation, identification and certain characteristics of a Hantaan virus proposed as candidate virus for killed vaccine]. AB - OBJECTIVE: Isolation and identification of a Hantaan virus from HFRS patient's serum for killed vaccine development. METHODS: This virus temporally called FT14 detected by IFAT, RT-PCR. Partial sequence of M segment was determined by Sanger' a dideoxy chain termination method. RESULTS: The antigenicity and PCR amplification of the newly isolated virus FT14 proved that it belongs to virus of HTN type. Antiserum to the virus can neutralize many strains of Hantaan virus from different areas. The partial sequence of M segment of FT14, as compared with other HTN strains, showed great differences among nucleotide sequences, while the deduced amino acid sequence is comparatively conservative. The titer of the virus reached its peak at 9 days post inoculation in vitro, then the peak titer could last for a long time. CONCLUSIONS: This virus strain FT14 would be a good candidate virus for killed vaccine development. PMID- 11503021 TI - [Study on relationship between the apoptosis of hepatocytes and liver fibrosis of chronic viral hepatitis]. AB - OBJECTIVE: To study the relationship between apoptosis of hepatocytes and liver fibrosis of chronic viral hepatitis (CH). METHODS: DNA fragmentation, one of the characteristic features of apoptosis, was detected by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and the expression of Fas, transforming growth factor beta 1 (TGF-beta 1) and Pre collagen III peptide (P III P) in liver tissues were detected by immunohistochemistry. Soluble Fas (sFas) and TGF-beta 1 in serum were detected by ELISA. RESULTS: The damage of DNA in hepatocytes of CH correlated closely with the expression of Fas and TGF-beta 1 in liver tissue and serum (r = 0.6129, 0.5368, 0.5564, 0.5996; P <0.05-0.01). In severe type CH and liver cirrhosis, the degree of liver fibrosis and the expression of P III P and TGF-beta 1 were higher than that in mild type and moderate type CH. CONCLUSIONS: The sFas in serum may be one of the factors associated with the apoptosis. The apoptosis of hepatocytes correlated with liver fibrosis of CH and TGF-V beta 1 may play an important role between apoptosis and liver fibrosis. PMID- 11503022 TI - [Study on the origin of influenza A(H1N2) virus HA and NA genes]. AB - OBJECTIVE: To determine the origins of the HA and NA genes of new subtype (H1N2) of influenza A virus. METHODS: Virus was amplified in embryonated chicken eggs, then virion RNA was transcribed into cDNA by reverse transcriptase, cDNA amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method, using synthetic oligodideoxynucleotide primers. Finally, phylogenetic analysis of the sequencing data was per-formed with MegAlign (Version 1.03) and Editseq (Version 3.69) software. RESULTS: The homology of amino acid sequences of protein molecules on HAl domains when compared, with A/PR/8/34(H1N1) and A/Guangdong/6/91(H1 N1) viruses was 98.2% and 99.4%, respectively. The homology of amino acid sequences of NA protein molecules between the novel reassortant and H1N1 virus was only 83.4 %, but the homology of those between H1N2 strain and H3N2 virus circulating in men in 1995 was as high as 99.1 %. CONCLUSIONS: The new reassortant (H1N2) virus HA gene was derived from A/PR/8/34(H1N1) like virus, but its NA gene was derived from H3N2 virus circulating in men in 1995. PMID- 11503023 TI - [Viral etiology study of acute conjunctivitis]. AB - OBJECTIVE: To study the viral etiology of acute conjunctivitis broke out in Shenzhen in 1997. METHODS: Isolated the virus from eye and pharynx swabs of acute conjunctivitis in acute stage, and detected virus nucleic acid by polymerase chain reaction (PCR). DNA of viral strains were subjected to restriction enzyme analysis using several endonucleases. Also, neutralizing antibody of paired sera were detected. RESULTS: The positive virus isolation rates of 14 pharynx and eye swabs were 71.43% and 64.29%, respectively; and that of the virus DNA by PCR were 78.57% and 71.43%, respectively. All of the strains isolated were adenovirus type 3. Three paired viral strains chosen and subjected to restriction enzyme analysis proved belonging to Ad3a2 gene type. The neutralization test of paired sera of 24 cases showed that the antibody titers had 4 fold rising or above. Neutralization test of isolated viruses with paired sera of itself was carried out, the level of neutralizing antibody in convalescent stage was above 4 times as compared with that of acute stage. CONCLUSIONS: The results showed that the outbreak of acute conjunctivitis occurred in 1997 in Shenzhen was caused by adenovirus type 3. PMID- 11503024 TI - [Antibody to human herpesvirus type-8 in the general populations of Xinjiang Autonomous Region(A.R.)]. AB - OBJECTIVE: Kaposi 's sarcoma (KS) does not rarely occur in Xinjiang A. R. Much of the evidence show that human herpes virus-8 (HHV-8) is associated with KS, and the different incidence of classic KS among different countries reflects the different prevalence in general populations. The aim of this study was to know whether the prevalence of HHV-8 is higher in general populations of Xinjiang than the other places of the country. METHODS: We used BCBL-1 cell line as antigen and collected 1071 serum sample of different nationalities (Uighur, Khalkhas, Kazak and Han nationality) from south and north regions of Xinjiang A. R. for detection of HHV-8 IgG antibody by IFA. RESULTS: The results showed that HHV-8 IgG antibody positive rate in Khalkhas was 48% (96/200), Uighur 30.4%(62/204), Kazak 12.5% (25/200) and Han 16.9% (79/467). While sera were diluted in 1:20, the positive rate in Khalkhas still was as high as 30.5% (61/200). In addition, the positive rate of HHV-8 IgG antibody in minorities in the same native area all were higher than Han except Kazak nationality. CONCLUSIONS: These data illustrated that the prevalence of HHV-8 IgG antibody in general populations of different nationality was different, Khalkhas and Uighur were higher than Kazak and Han. Most of the KS cases were reported from Uighur nationality, but no case was reported from Khalkhas thought a highest HHV-8 IgG antibody detection rate (48%) there was demonstrated. The reason for it would be further studied. PMID- 11503025 TI - [Detection of hepatitis C virus RNA in the tissue of hepatocellular carcinoma by multiple detection system]. AB - OBJECTIVE: Detection of hepatic HCV RNA in hepatocellular carcinoma (HCC) is difficult, since its expression is very low. Several techniques have been established. However, false positive and negative rates still exist. In this study, we applied several conventional and recently developed detection systems to determine the exact effect of HCV RNA on the development of RCC. METHODS: Immunohistochemistry for HCV core antigen and in situ hybridization for HCV RNA had been performed in 39 cases of HCC. IS-RT-PCR was applied for detection and localization of HCV RNA. We extracted microdissected liver tissues to detect HCV RNA separately in cancerous and pericancerous tissues so that the microscopic origin of the amplicons could be controlled. The serological tests including ELISA and RT-PCR for HCV also were performed in all cases. RESULTS: The positive rate by ELISA (30.8%) was not always consistent with that by RT-PCR(43.6%)from serum samples. The latter was much more sensitive and accurate to reflect existence of HCV RNA. Immunohistochemistry showed expression of RCV core antigen in 15 of 39 HCC cases. In pericancerous tissues, the signals were mainly localized in the cytoplasm of hepatocytes. However, translocated expression of HCV core protein was observed in the nuclei of tumor cells, the translocated rate was 73.3%. According to IS-RT-PCR, the positive signals were located mainly in the cytoplasm of cancer cells, the positive rate in HCC was 53.8%, expression of HCV RNA, serum HCV RNA level was detected to be low or negative. This suggested that serum HCV RNA was not always a good reflection of hepatic HCV RNA in HCC tissues. The microdissection RT-PCR described here gave an equal positive rate (59.0%) of HCV RNA in cancerous and pericancerous tissues. CONCLUSIONS: The high detection rate of HCV RNA in HCC specimens even from seronegative patients confirms the important role of HCV RNA during malignant transformation. IS-RT-PCR is a good detection and localization method to visualize HCV RNA in HCC tissues. The microdissection RT-PCR method has a distinct advantage and we anticipate this method will provide an important evidence to determine whether HCV play a direct or indirect role in hepatocarcinogenesis. PMID- 11503026 TI - [Photodynamic inactivation of model virus in human plasma by methylene blue and its influence on plasma proteins]. AB - OBJECTIVE: To determine whether photochemical method can inactivate viruses in human plasma efficiently, and to what extent it affects the protein in human plasma. METHODS: Vascular stomatitis virus (VSV) Indiana strain was used as the indicating virus. It was inoculated on Vero cell monolayer, incubated in 37 degrees C for 24 hours, microtitered by cytopathogenic effect. Small amount experiments were carried out to observe virus inactivation and the influence on plasma proteins. RESULTS: Methylene blue (MB) in combination with visible light can inactivate the model virus in human plasma efficiently. The dye in very low concentration combined with light was sufficient to achieve a reduction of VSV titer of more than 6 log10 TCID50. Compared with the untreated plasma, neither the activities of blood clotting factors nor the immunological properties of the plasma proteins and the blood albumin were reduced significantly. In this experiment, the dose of MB used for photoinactivation of the virus was very low, about a hundred fold lower than that of methemoglobinemia used in clinical treatment. Furthermore, heat of the light and photochemical reaction are two import factors for decreasing of clotting factor's activity. CONCLUSIONS: The results of this preliminary study suggest the potential application of MB photodynamic inactivation of plasma virus is an useful approach in clinical practice. PMID- 11503027 TI - [Mechanism of Se-Indocalamus tessdatus polycassine(S-ITPS) against human immunodeficiency virus type 1 (HIV-1)]. AB - OBJECTIVE: To study the mechanism of Se- Indochalamus tessdatus polycassine (S ITPS) against HIV-1 and to provide a theoretical support for developing the anti HIV drugs. METHODS: S-ITPS was added to MT4 cells before viral inoculation, at the same time of viral inoculation, then the cells were cultured with or without S-ITPS for 1-4 weeks, finally, cytopathic effect (CPE), MTT staining method for viable cells(MTT assay),p24 antigen titer (ELISA), or infectivity (TCID50 assay) of the cultural supernatants were used as markers to monitor the virus growth. RESULTS: S-ITPS can inhibit HIV-induced CPE (1C50 = l0microg/ml)and viral replication (1C50 = 156 microg/ml)in dose dependent manner. The virus infectivity in the presence of S-ITPS was greatly decreased than that of the control. The virus inhibition was enhanced under the presence of noncytotoxic drug concentration of <1250 microg/ml in cell culture, inhibition of viral replication by S-ITPS was 73.7%, 63.5%, 87.7% and 95.4% at week 1, week 2, week 3 and week 4 of cultured cells respectively. It can also inhibit absorption of HIV-1 to MT-4 cells, but no inhibition of HIV-1 was seen when MT-4 cells were pretreated with S ITPS. Virus can be inactivated by the agent also. CONCLUSIONS: S-ITPS is a potent anti-HIV- 1 agent in MT-4 /HIV- 1 cultural system, at least two mechanisms were included inhibition both of HIV-1 absorption to MT-4 cells and viral replication in HIV-infected cells. S-ITPS inhibition of CPE is stronger than that of p24 antigen production. PMID- 11503028 TI - [Cloning and expression of full-length subtype E and subtype B gag gene from uncultured PBMCs of HIV-1 infected individuals in China]. AB - OBJECTIVE: To clone and express subtype E and subtype B gag gene of the prevalent HIV-1 strain in China. METHODS: One HIV-1 positive sample, whose subtype had been determined as sub-type E and one HIV-1 positive sample, whose subtype had been determined as subtype B by sequence analysis of partial env gene were chosen to be amplified by nested PCR. We got two full-length fragments of gag gene and incorporated them first into plin8Pr55 and then into pFastBacl. RESULTS: Sequence analysis showed the cloned gag genes have the complete open-reading frames and have no major deletion and insertion. Using the Bac-toBac system, we obtained the recombinant baculoviruses containing gag genes. Western-blot analysis showed that insect cells infected with gag recombinant baculoviruses expressed HIV-1 gag antigen. Thin section electron microscopy showed that virus-like particles (VLPs) formed in insect cells was infected with gag. CONCLUSIONS: The cloning and expression of gag genes of the prevalent HIV-1 stains in China revealed that the cloned genes have intact structure and function and this paves the way for the development of HIV-1 vaccines targeted at the epidemic of HIV-1 in China, also for the study of biological functions of gag gene and development of diagnostic kits of HIV-1. PMID- 11503029 TI - [Cloning and sequencing of complete TT virus genome from Beijing isolate of China]. AB - OBJECTIVE: To obtain the complete TT virus genome from a Beijing isolate of China. METHODS: PCR was performed for amplifying the TT virus genome from serum of a patient with non A-G hepatitis. Five overlapped fragments, that are 199 bp (1-199), 1 267 bp (90-1356), 878 bp (1354-2231),1129 bp(2178-3306),'434 bp(3306 3739), respectively, were obtained. These PCR products were sequenced by standard method. RESULTS: The complete TT virus genome of Beijing isolate consists of 3 739 nucleotides involving two ORFs. The first ORF locates at 589-2902 nt, encoding 770 amino acids, the second ORF locates at 107-715 nt, encoding 202 amino acids. 5' and 3' terminus of the genome are non-coding regions including 106 bp and 837 bp respectively. In this genome, A is 31.2%, C 25.4%, G 22.5% and T 20.9%. The homology of Beijing isolate and Japanese TT virus was more than 95% at both nucleotide and amino acid levels. CONCLUSIONS: We successfully cloned and sequenced the complete TT virus genome from a Chinese patient. This Beijing TT virus isolate and that reported by Japan all belong to a same genotype. PMID- 11503030 TI - [Expression, purification and identification of human IFN-alpha 2 b/HBV Pre S2 fusion protein]. AB - OBJECTIVE: To express a fusion protein of human interferon-a2b and HBV Pre S2 in E.coli for the purpose of investigating anti-HBV immunomodulatory protein. METHODS: Human interferon-a2b and HBV Pre S2 encoding genes were amplified from plasmid templates through PCR, then fused and cloned into plasmid pBV220 through engineering technique to generate expression plasmid pBV-IFN-Pre S2. The plasmid was transfected into E. coli to produce fusion protein. RP-HPLC and ion-exchange chromatography were employed to purify fusion protein. RESULTS: 27 kDa fusion protein was expressed up to 15% of total bacterial protein in E. coli. After purification, the purity of fusion protein reached 95% of total protein. Anti viral assay showed that IFN-Pre S2 protein induced a VSV-resistant activity of 1.25 x 10(8) lU/mg protein in Wish cell line, similar to the bioactivity of original recombinant human IFN-alpha 2b. ELISA data showed that IFN-Pre S2 protein had antigenicities of both human IFN-alpha 2b and HBV Pre S2. In addition, fusion protein showed the feature of binding to polymeric human serum albumin (PHSA). CONCLUSIONS: The bifunctional fusion protein was efficiently expressed in E. coli. The work provided initial evidence for studying PHSA receptor-targeting IFN-alpha 2b,which might have potential application for the treatment of HBV infection. PMID- 11503031 TI - [Establishment and application of Z-HL16C cell line]. AB - OBJECTIVE: Human embryonic lung fibroblasts at 8th passage was presented to us by the Institute of Virology. At its 16th passage,under the culture condition of our laboratory, the lung fibroblasts changed into epithelial like cells. We attempted to establish a cell line and applied it for virus isolation. METHODS: The cells were continuously passaged, cloned, the cell morphology, growth capacity and chromosome number were determined. RESULTS: The cell size appeared uniform, cell boundary was distinct, the cell recovery rate after frozen storing was above 90%. The cell replicated permanently and now it has been passaged 136 times. The chromosome number has changed from 46 to 110. We named this cell line the ZHL16C. It proved to be sensitive to these viruses tested: enteroviruses (Polio, Cox, Echo), influenza viruses, parainfluenza viruses, adenoviruses, measles virus, herpes simplex and herpes zoster viruses, cytomegalovirus, rubella virus and respiratory syncytial virus. When using Z-HL16C cell to isolate virus from 29 adenovirus swab samples collected from 29 soldiers with epidemic high fever in a new military, adenovirus type 3,7 coxsackie virus types B1, B5, B5, B5, B5 were isolated. CONCLUSIONS: The cell line HL16C has been stably established, it has a broad spectrum in sensitivity for viruses. PMID- 11503032 TI - [A prospective study of posttransfusion hepatitis B virus infection]. AB - OBJECTIVE: To understand the status of HBV posttransfusion infection after blood HBsAg screening of the blood donors. METHODS: Serum samples of blood recipients and donors were detected for HBV-M and HBV DNA by ELISA technique and polymerase chain reaction respectively. RESULTS: There were 326 sera positive for HBV-M and 57 for HBV DNA in 583 blood donors' sera. 39 sera were positive for HBV-M and 23 positive for HBV-DNA in 136 sera of blood recipients, the positive seroconversion rates were 28.68% and 16.9% respectively. CONCLUSIONS: There is high incidence of HBV infection in blood donors though they have been screened for HBsAg. The posttransfusion HBV infections are still occurring, therefore, the blood donors should be strictly tested for HBV-M. If conditions are possible, the HBV DNA had better been screened. PMID- 11503033 TI - [Preparation of hepatitis C virus nonstructural region antigens and detection of their relative antibodies]. AB - OBJECTIVE: To investigate the relationship between HCV NS3 and the pathogenesis of HCV and research a method to assess the therapeutic effect of interferon (lEN) in clinic. METHODS: In a retrospective research, we used HCV-NS3 antigens of different terminals expressed in E. coli which contained the recombinant plasmids involving amino terminal region (N') or carboxyl terminal region (C') of HCV-NS3, to detect the antibodies by Western blot in sera of 63 hepatitis C patients who were treated with IFN. RESULTS: It proved that antibodies against HCV-NS3 N' and/or HCV NS3 C' terminals in hepatitis C patients may predict the treatment result of IFN, especially in the patients with chronic hepatitis C. CONCLUSIONS: The detection of antibodies against HCV-NS3 N' and/or C' terminals may be used for evaluation of IFN treatment, especially in chronic hepatitis C ones. PMID- 11503035 TI - [The studies on human endogenous retrovirus gene expression, distribution of the genome and association with IDDM and other inheritance- related diseases]. AB - OBJECTIVE: The mechanism for type 1 diabetes still remains unclear. This study was focused on the viral morphology, endogenous retrovirus expression, immunology and molecular biology. METHODS: Type C retrovirus-like particles were observed in beta cells of the diabetic pancreases under electron microscope. Human endogenous retroviral pol gene expression was detected by PCR in cDNA libraries from IDDM pancreases, brain, splenocytes and control subjects. These DNA segments were sequenced. RESULTS: ERV9, HERV-K-MIN, HERV-K10, RTVL related sequences were significantly higher in IDDM group than control subjects and DR2-related MMTV like sequences were significant lower in IDDM group than control subjects. ERV9- related genes, gag, pol, env and 3' LTR were amplified and sequenced by PCR from cDNA libraries of 2 newly onset diabetic pancreases. Retrovirus sequences were further searched for their distribution on the chromosome genome and their locations at the centromeres, telomeres, immune reaction genes (MHC, TCR, Ig autoantigens) and at many disease loci, such as the loci for IDDM, Mody, the candidate genes for NIDDM, diabetic complications, autoimmune diseases, cancer and other genetic diseases. CONCLUSIONS: These facts indicate that the endogenous retrovirus genes may be the important genetic factor of IDDM and play key role in the structure and physical function of chromosome and the pathogenesis of genetic related diseases. PMID- 11503034 TI - [Detection of TT virus DNA by polymerase chain reaction-microplate hybridization]. AB - OBJECTIVE: To establish a method for the detection of TT virus (TTV) DNA by polymerase chain reaction (PCR)-microplate hybridization and to find out various influential factors of the assay. METHODS: TTV DNA extracted by guanidinium thiocyanate was amplified by PCR, the PCR primers were pre-labeled with biotin. By conjugation of biotin with streptavidin, the amplified products were coated to the microplate well, and hybridized with FITC- probe simultaneously. After being reacted with Anti-Fluorescein POD, the products that contained FITC probe were coloured by TMB-H2O2 and then measured the optical density using a microplate reader at 450 nm wavelength. RESULTS: The optimal concentration of NaOH was 0.3 mol/L, and the optimal time of hybridization and enzymatic reaction was 60 minutes and 45 minutes, respectively. The results also showed that the positive rate of TTV infection in normal population, hepatitis A-G and hepatitis non A-G was 13.6%, 20.3% and 28.1%, respectively. CONCLUSIONS: Comparing the PCR microplate hybridization with the electrophoresis assay, the former is simple, rapid, sensitive and specific and is expected to be a new technique instead of electrophoresis assay as a routine method for TTV DNA detection. PMID- 11503036 TI - [The partial nucleotide sequence of L segment of Hantavirus strain]. AB - OBJECTIVE: To study the nucleotide sequence of L segment of Hantavirus strain A9. METHODS: The partial L segment cDNA of A9 strain was amplified by PCR, the PCR product was sequenced. RESULTS: The nucleotide sequence of partial L segment of A9 strain was provided, the sequence was A, T rich, similar to other Hantaviruses, and similar to both M, S segments. Compared with nucleotide sequences from other Hantaviruses, it revealed a higher homology to HTN virus strain 76-118,83.5% at nucleotide level and 96.7% at deduced amino level. SNV, PUU and Tula viruses shared the lower homology. A9 and 76-118 were at same cluster in phylogenetic analysis, close to SEO virus but not to PUU, SNV, Tula viruses. Comparing of partial nucleotide sequence of strain A9 with 76-118, some conserved regions were found, but also some regions of A9 were more conserved to other viruses not 76-118. It suggested that when design the primers for L segments amplification, more than one type of Hantavirus sequence should he concerned. Like the homology from M and S segments of Hantaviruses, the viruses isolated from same host animal showed high homology. CONCLUSIONS: Nucleotide sequence of L segment of Hantavirus strain A9 revealed homology between 85.2% 70.8% to different types of Hantavirus and shared a higher homology of 85.2% to strain 76-118. PMID- 11503037 TI - [The effect of coxsackie virus B3 on intracellular calcium homeostasis in rat cardiomyocytes]. AB - OBJECTIVE: To investigate the effect of coxsackievirus B3 (CVB3) on intracellular calcium, L type voltage dependent calcium channel (VDCC) and Na+/Ca2+ exchange (NCE) in adult rat cardiomyocytes. METHODS: Rat hearts were perfused with collagenase to gain single cardiomyocyte. The intracellular free calcium concentration of the cardiomyocytes were detected by laser confocal microscopy with calcium indicator, Fluo3-acetoxymethyl ester; and the currents of VDCC, Na+ and NCE were recorded by patch clamp techniques. RESULTS: The free intracellular calcium concentration increased from (51 +/- 43) nmo1/L in normal cardiomyocytes to (339 +/- 208) nmo1/L in CVB3 infected cardiomyocytes. The VDCC current increased from (4.13 +/- 0.75) pA/pF to (4.9 +/- 1.4) pA/pF after CVB3 infection, and the inward NEC current decreased from (-0.46 +/- 0.21) pA/pF to (-0.28 +/- 0.16) pA/pF; besides, the reverse potential of NCE was shifted from (29.4 +/- 7.0) mV to (41 +/- 14) mV with CVB3 infection. CONCLUSIONS: The increase of L type VDCC current and decrease of inward NCE current with shift of NCE reverse potential may partly attribute to intracellular free calcium accumulation; also, it may be the cause of cell damage and the occurrence of abnormal electric activities induced by CVB3 infection. PMID- 11503038 TI - [Study on the carcinogenic mechanism of human papillomavirus typel6 E7 protein in cervical carcinoma]. AB - OBJECTIVE: To determine the relationship between HPVl6 E7-Rb protein binding (Rb protein was dissociated from pRb-E2F-l complex) and the occurrence of cervical carcinoma. METHODS: HPV16 DNA infection rate was tested in cervical carcinoma and normal cervical tissues by polymerase chain reaction (PCR) technique. Then using Western blot analysis to find the presence HPVl6E7 protein and pRb-E2F-1 complex in HPV16 DNA positive cervical carcinoma tissues. The normal cervical tissues were used as control. RESULTS: Out of 67 samples with cervical carcinomas, 34 samples were HPV16 DNA positive (50.7%), no HPVl6 DNA were detected in 10 samples of normal cervical tissues. In cervical carcinoma tissues, the HPV16E7-pRb complex, the free E2F-1 were detected by Western blot, but no complex of pRb-E2F l was found. CONCLUSIONS: Our results seem to suggest that HPV 16 E7 protein could interact with pRb through separating the complex of pRb-E2F-1, release free E2F- 1, resulted in disordered cell cycle and cell malignant transformation. PMID- 11503039 TI - [Generation of monoclonal antibody Fab fragments to parathyroid hormone related protein by phage display technology]. AB - OBJECTIVE: To develop monoclonal antibody Fab fragments using display technique. METHODS: The mouse IgG Fab genes of heavy and light chains were amplified from spleen cells of a parathyroid hormone-related protein (PTHrP) immunized mouse. The combinatorial phage antibody library was prepared by inserting both heavy and light chain Fab genes into phagemid vector pComb3 and followed by infection of helper phage. The library was selected by purified recombinant PTHrP. RESULTS: The combinatorial phage antibody library was constructed successfully and the specific mouse Fabs to PThrP were selected and expressed in E. coli. CONCLUSIONS: The selected specific mouse Fabs can recognize PTHrP with high specificity. PMID- 11503040 TI - [The efficacy of antiviral antibiotic 17997 on treatment of HSV-1 infected guinea pig skin infection]. AB - OBJECTIVE: To study the treatment efficacy of antiviral antibiotic 17997 against HSVl infected guinea pig skin infection. METHODS: Guinea pig skin was infected by HSV1. 24hrs or 48hrs of post infection local treatment of 0.3% 17997 cream was started, tid for five days. In the mean time, acyclovir treatment, cream treatment and virus control were included. RESULTS: Local treatment of 0.3% 17997 cream showed therapeutic effects, it reduced the average scores of skin lesion, accelerated crusting-time and healing-time. CONCLUSIONS: 0.3% 17997 cream showed significant treatment efficacy when compared with cream and virus controls by reducing skin lesion scores and healing-time. The treatment efficacy of 3.0% acyclovir cream was a little bit better than 0.3% 17997 cream. PMID- 11503041 TI - [Inhibition of the expression of Epstein-Barr virus antigens in vitro by Chinese medicine compound "Dongxia wan"]. AB - OBJECTIVE: To investigate the effect of Chinese medicine "Dongxia wan" on the expression of Epstein-Barr virus (EBV) antigens in vitro. METHODS: Cell culture and indirect immunoenzyme methods were used. RESULTS: When Raji and B95-8 cells were cultured with 20-80 microg/'ml of "Dongxia wan", the expression of EBV early antigen (EBV-EA) and EBV capsid antigen (EBV-VCA) induced by croton oil and n butyric acid and the natural expression of EBV-VCA were all inhibited significantly. The inhibition rates were 19.3% - 49.84%, 34.63% - 45.61% and 21.67% - 47.78%, respectively. When Raji and B95-8 cells were pretreated with "Dongxia wan" the expression of EBV-EA and EBV-VCA were also inhibited obviously. CONCLUSIONS: Chinese medicine "Dongxia wan" can inhibit the expression of EBV antigens in target cells, it may be used to the prevention and treatment of EBV related diseases. PMID- 11503042 TI - [Preparation of polyclonal antibody to human prion protein using the expressed GST-PrP fusion protein as antigen]. AB - OBJECTIVE: Preparing specific antibody to prion protein. METHODS: Using prokaryotic expressed GST-PrP fusion protein as antigen, rabbits were immunized subcutaneously. RESULTS: ELISA assay revealed that the titer of the prepared antiserum against expressed PrP protein was as high as 1:128 000. Western blot test showed that the antiserum was able to react with the in vitro expressed intact and different lengths of C-terminus truncated PrP proteins, as well as the native PrP proteins from brain homogenization of human and mouse. CONCLUSIONS: The prokaryotic expressed GST-PrP fusion protein can efficiently elicit in immunized animals the PrP-specific antibody. PMID- 11503043 TI - [Further study on the genetic characteristics of a strain of A/PR/8/34/]. AB - OBJECTIVE: Through the test and analysis of nucleotide sequences of seven different RNA segments of the virion to exclude the possibility that A/Guangdong/6/91 (H1N1) strain might be derived from laboratory contamination and to determine how many RNA segments are there with nucleotide sequence difference between A/Guangdong/6/91 (H1N1) and A/PR/8/34 (H1N1) viruses, as well as to determine is A/Guangdong/6/91 virus a reassortant? METHODS: Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA was amplified by PCR, the products of PCR were purified. Afterward, RNA sequence test was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers. RESULTS: There are no nucleotide sequence differences in RNA segments 6 and 7 between A/Guangdong/6/91 (H1N1) and PR8 strains, while there are 20,5, 11, 7 and 6 differences in nucleotide sequences of RNA segments 1, 2, 3, 5 and 8 between these two strains, respectively. These differences induce 10, 2, 1, 0 and 4 differences in amino acid sequences on PB2, PB1, PA,NP and NS protein molecules between A/Guangdong/6/91 (H1N1) and PR8 strains, respectively. CONCLUSIONS: The possibility of laboratory contamination of A/Guangdong/6/91 (H1N1) virus can be ruled out completely. A/Guangdong/6/91 (H1N1) virus is a PR8 like strain, but not a reassortant. PMID- 11503044 TI - [Effect of Sophora flavescens Ait on cultured beating myocardial cells of coxsackie B3 virus infected newborn rat]. AB - OBJECTIVE: Coxsackie B viruses (CVB3) are considered to be the most common etiologic agents of viral myocarditis. There are not any special anti-CVB3 drugs yet. From previous studies, the anti-CVB3 affect of sophora flavescens ait (SFA) had been discovered. Our experiment was to study the anti-CVB3 ability of SFA to cultured heating myocardial cells of CVB3 infected newborn rat. METHODS: The myocardial cells were divided into four groups: 1. infected group (n = 8), infected only with CVB3, not adding SFA; 2. treated group (n =8), infected with CVB3, adding SFA (100 microg/ml); 3. drug group (n=6), adding SFA (100 microg/ml) only; 4. control group (n = 6), not infected with CVB3, not adding SFA. RESULTS: The myocardial cells of the infected group had cell pathogenic effect (CPE) on the second day after virus inoculation, the CPE progressed rapidly from + to ++++. In contrast, no CPE in the other three groups was found. The LDH and SGOT of the infected group were higher than that in the other three groups, showing a significant difference (P <0.05). The virus titer of the infected group was higher than that of the treated group. There was no influence on normal myocardial cells if the concentration of SFA was lower than 300 microg/ml. When the concentration of SFA was 6.25 microg/ml 200 microg/ml, it showed protective effect on infected myocardial cells. CONCLUSIONS: The results of the experiment suggest that SFA might inhibit CVB3 replication in myocardial cells. PMID- 11503045 TI - [Molecular design and immunogenicity studies of multiple antigenic peptide corresponding to envelope glycoprotein hypervariable region 1 of hepatitis C virus]. AB - OBJECTIVE: To study the immunogenicity of multiple antigen peptide (MAP) corresponding to the hypervariable region 1 (HVR1) within the putative envelope glycoprotein E2/NS1 of hepatitis C virus (HCV). METHODS: Basing upon the E2/NS1 consensus sequence of HCV-BJ (isolate of Beijing) which we had obtained previously, and referring to several HCV HVR1 sequences reported by foreign authors, we synthesized a linear epitope peptide (LP) and a MAP (symmetric 8 branches) according to 22 amino acids (position 390 - 411 aa) of HCV-BJ HVR1 on the basis of our prediction to their antigenicity parameters. Then, the Balb/C mice and rabbits were immunized with IP or MAP respectively. We compared the immunogenicity of MAP with Lp. RESULTS: The results showed that the immunogenicity of MAP which elicited high-titer antibody more than 1:40 960 was much stronger than that of LP (1:1 280). CONCLUSIONS: It suggests that the immunogenicity of peptide corresponding to HCV HVR1 was stronger by using MAP conformation, and this provide a basis for designing HCV molecular vaccine using MAP. PMID- 11503046 TI - [Determination of serum cytokines in individuals with HCV infection]. AB - OBJECTIVE: To investigate the immunopathogenesis of chronic hepatitis C virus (HCV) infection. METHODS: Serum T-helper lymphocyte (Th1) cytokines (IFN-gamma, IL-2) and Th2 cytokines (IL-4, IL-10) were measured by ELISA in 18 individuals with chronic HCV infection 10 persons of chronic HBV infection and 11 normal persons. RESULTS: IL-2 (P <0.05), IL-4 and IL-10 (P <0.025, P <0.001) were significantly increased in HCV infected hosts as compared to 11 normal controls, the production of Th2 cytokines was more predominant. CONCLUSIONS: An enhanced Th2 responses is present in HCV infection, which may he responsible for the chronicity of HCV infection. PMID- 11503047 TI - [Epidemiological aspects of the genotype of hepatitis C virus]. AB - OBJECTIVE: To research the epidemiological characteristics of genotype of hepatitis C virus. METHODS: 107 HCV isolates from 7 cities in southern, northern and northeast of China were analyzed using PCR and second generation line probe assay (INNO-LiPA). RESULTS: The analysis of HCV genotypes showed that: 1. HCV l b was the common genotype (83 17%), however the frequency of genotype l b in northeast (72.22%) was lower than northern (87.65%, P <0.01), and the frequency of the genotype 2 was in low level (6.86%). The rate of coinfection with l b and other genotype was 10.29%. 2. The rate of infection with genotype l b in female patients was higher (93.61%) than that in male (75.0%) showing statistical significance (P <0.01). 3. In three age groups (< 30, 30 - 49 and > 50 years old) the rates of infection with genotype l b were 77.77%, 83.92% and 90.90%, respectively. 4. The detection rates of genotype l b in patients with infection period > 10 years and <10 years were 100% and 80.43% respectively (P <0.01). CONCLUSIONS: There are some relationship between HCV genotype and HCV geographic distribution, sex, age and infection period of patients. PMID- 11503048 TI - [Detection of IgG antibody to hepatitis G virus by ELISA]. AB - OBJECTIVE: To study the relation of HGV IgG antibody (anti- HGV IgG) to various viral infections, ALT and HGV RNA. METHODS: To detect anti-HGV IgG from 315 persons infected with various viruses hepatitis (A - E) and 117 healthy blood donors by ELISA. The detection of ALT of these peoples, and detect HGV RNA of the samples of positive anti-HGV IgG by RT-PCR were also carried out. RESULTS: The positive rate of anti-HGV IgG of 117 blood donors was 3.42% (4/117), and that of the people infected by non A- E, HAV, HBV, HCV, HEV, HBV and HCV, HBV and HDV, HBV and HEV were 5.95%(5/84), 0(0/3), 3.75% (3/80), 17. 28%(14/81), 0(0/15), 33.33%(5/15), 3.33%(1/30), 16.66%(1/6), respectively. The relativity of anti-HGV IgG and HGV RNA was 30.30% (10/33), and 33.33% of ALT level in the antibody positive samples were elevated. CONCLUSIONS: HGV may infect people alone, or coinfect with HBV or /and HCV, but is highly relative to HCV, and the coinfection of HGV with HCV may increase the degree of liver damage. PMID- 11503049 TI - [The study on hepatitis G virus infection among patients with liver diseases]. AB - OBJECTIVE: To investigate the hepatitis G virus (HGV) infection among patients with liver diseases. METHODS: Anti-HGV was detected by ELISA among 154 patients with liver diseases (no history of blood transfusion), 54 of them, HGV RNA was detected by RT-PCR concomitantly. The rates of anti-HGV and HGV RNA were calculated, respectively. The positivity of HGV RNA was taken as diagnostic criteria for HGV infection in different kinds of liver disease. RESULTS: Of 154 patients with liver diseases, anti-HGV was positive in 31 patients, the detectable rate of anti- HGV was 20.13%. HGV RNA was detected in 13 of 54 patients, the detectable rate was 24.07%. The positive rates of HGV RNA tested in 35 patients with chronic hepatitis and 6 patients with fatty-liver were 20.00% (7/35) and a ratio of 5/6, respectively. CONCLUSIONS: (1) HGV infection is considerably prevalent among patients with non-transfusion transmitted sporadic hepatitis. (2) HGV is a responsible agent for chronic hepatitis. (3) The clinical significance of detectable HGV RNA in patients with fatty-liver remains for further study. PMID- 11503050 TI - [The expression of P21 and P15 proteins in liver tissue of chronic viral hepatitis and hepatocellular carcinoma and their relation to cell apoptosis]. AB - OBJECTIVE: To study the expression of P21 and P15 proteins in liver tissue of chronic viral hepatitis (CH) and hepatocellular carcinoma (HCC) and their relation to cell apoptosis. METHODS: The expression of P21 and P15 proteins in 36 CH patients liver tissues were detected by immunohistochemistry and the DNA damage of the hepatocytes were detected by TDT mediated dUDT nick and labeling (TUNEL). 10 cases of HCC were also studied. RESULTS: P21 and P15 proteins mainly distributed in the interstitium of portal areas, the piece meal necrotic sites and the nuclei and cytoplasm of the hepatocytes. The higher expression of P21 and P15 proteins in the interstitium and nuclei of the hepatocytes was seen in CH as compared with that in HCC. The levels of expression of P21 and P15 in moderate and severe type CH were higher with that of the mild type CH (P <0.05). The correlative coefficient of expression of P21 and P15 with the DNA damage of hepatocytes were 0.56, 0.51 respectively (P <0.01). CONCLUSIONS: The enhanced expression of P21 and P15 proteins in CH correlated with the injury of liver tissues and apoptosis of hepatocytes. PMID- 11503051 TI - [The characteristics of clinical course of chronic hepatitis B and liver cirrhosis pathology of child patient]. AB - OBJECTIVE: To study the characteristics of clinical course of chronic hepatitis B and liver cirrhosis pathology of the child patients. METHODS: The serum test for hepatitis A/B/C/E, the liver function test and the liver puncture for pathological examination were performed in 66 child patients with chronic hepatitis B. RESULTS: The age, clinical course and form of disease onset, showed no-obvious difference between S0 group (without liver cirrhosis) and group > or = S3 (with evident liver cirrhosis). The cases with splenomegaly accounted for 2.9% in S0 group, but 32.3% in group > or = S3. The Hbe Ag positive rate of group S0 was apparently higher than that of group > or = S3 (74.29% and 29.03%, respectively). The higher the activity of liver inflammation, the more serious the extent of liver cirrhosis. CONCLUSIONS: The liver cirrhosis of child patients with chronic hepatitis B has nothing to do with their age, clinical course, form of disease onset and virus replication, but it is in direct connection with the liver inflammation activity. If splenomegaly occurs and thrombin acts abnormally, albumin in serum decreases while gamma-globulin increases, the liver cirrhosis should be highly watched out for. PMID- 11503052 TI - [Detection of core promoter mutations in chronic hepatitis B]. AB - OBJECTIVE: To study the core promoter mutation in chronic hepatitis B and its effect on viral serology. METHODS: HBV core promoter gene fragments were amplified by using mismatched PCR combined with a restriction fragment length polymorphism assay. The PCR products were digested with Bcl I and subjected to electrophoresis on agarose gels. RESULTS: We investigated the core promoter mutation in 89 patients with HBV DNA positive. The patterns of restriction fragment length polymorphism (RFIP) of the core promoter gene were distinguished and verified by direct sequencing. The combined mutations of nucleotides (nt) 1762 and 1764 in the core promoter from A to T and G to A were detected in 47 individuals, 21 of 43 HBeAg positive patients and 26 of 46 anti- HBe positive cases were found infected with this combined mutant. These two groups showed no significant difference (P > 0.05). The combined mutation was found in 11 of the 15 patients with medium and severe chronic hepatitis. In 5 chronic hepatitis B patients with coexistence of wild-type and mutant a tendency of superior accumulation of mutant was found. CONCLUSIONS: This study suggests that the core promoter mutations commonly exist in Chinese chronic hepatitis B patients. The mutation can only reduce the pre-core mRNA transcribing efficiency, but cannot discontinue the synthesis of HBeAg. The effect on serology in this mutation is different from that in pre-core stop 28 mutation. The superior accumulation of mutations seems relating to the degree of chronic liver disease. PMID- 11503053 TI - [The molecular epidemiology of transfusion transmitted virus infection in Shanxi Province]. AB - OBJECTIVE: To study the molecular epidemiology of transfusion transmitted virus (TTV) infection and its distribution in different populations in Shanxi Province and the gene types of the isolates. METHODS: TTV DNA in serum samples was detected by polymerase chain reaction. Partial gene of the strains from different populations were cloned and sequenced, and their gene types were analyzed. RESULTS: Of 24 cases with chronic hepatitis B, 31 cases with non A-G hepatitis, 53 cases of professional blood donors, 112 cases of healthy lying -in women, the positive rates of TTV DNA were 41.66%, 29.03%, 58.49% and 8.93%, respectively. TTV DNA in serum samples of 18 cases of 2 - 3 years old healthy children were negative and also were negative in umbilical cord blood of 10 cases of TTV DNA positive healthy lying-in woman. Sequence analysis of 4 isolated strains of Shanxi Province showed that the nucleotide identity among them were more than 90% and they all belonged to the same gene type (la) CONCLUSIONS: This study proved that TTV infection existed in Shanxi Province, TTV might be an important pathogen for non-A to G hepatitis. TTV often coinfected with hepatitis B virus and there were high rates of TTV infection in professional blood donors and in patients with chronic liver disease. The gene type of TTV in Shanxi province was la. PMID- 11503054 TI - [High prevalence of TT virus (TTV) infection and gene subtype analysis between Japanese and Chinese]. AB - OBJECTIVE: To compare the infection rates and gene subtypes of TTV between Japanese and Chinese. METHODS: TTV was detected by a semi-nested PCR. The positive samples were sequenced directly. RESULTS: Infection rate of T'TV in normal Chinese was 64%, but in the patients suffered from chronic C hepatitis, the infection rates were 68% in Chinese and 49.1% in Japanese The gene subtype of TTV Gla + 1b in Chinese was 40%, G2a+ 2b 60%; in Japanese TTV Gla+ 1b was 78.6%, G2a+ 2b 21.4%. CONCLUSIONS: There were high infection rates of TTV in Chinese and Japanese. The gene subtype distribution of TTV had some difference between Chinese and Japanese. PMID- 11503055 TI - [Therapeutic effect of blood activating polypeptide on the treatment of chronic severe viral hepatitis]. AB - OBJECTIVE: To study the therapeutic effect of blood activating polypeptide on chronic severe viral hepatitis (CSH) and the method used. METHODS: The treated group included 50 CSH patients (male 82, female 8, age 39 +/- 9). Based on routine treatments, they were intravenously injected with 15 ml blood activating polypeptide dissolved in 250 ml of 10% glucose, qd x 4 wks. The control group contained 50 CSH patients (male 3, female 7, age 38+/-12). They were only treated with routine treatments for liver protection, jaundice regression, health maintenance, anti-infection and diuresis. RESULTS: The SB and ALT in treated patients decreased obviously (P < 0.01) after treatment, the prominent efficacy was 52%, the total efficacy was 86%, all were superior to the control group. CONCLUSIONS: Blood activating polypeptide is safe, reliable and effective in the treatment of CSH. PMID- 11503056 TI - [The localization of parvovirus B19 in cardiac biopsy tissue of congenital heart disease]. AB - OBJECTIVE: To explore the relationship between parvovirus B19 infection and congenital heart disease (CHD), and distribution of parvovirus B 19 gene in cardiac tissue. METHODS: We conducted a case controls study investigating the presence of B19 gene in cardiac biopsy tissue of 37 cases of CHD, 8 cases of rheumatic heart disease (RHD) and 20 cases of non-CHD with nested polymerase chain reaction (Nested PCR) and in situ hybridization (ISH). RESULTS: Among 37 CHD patients, 6 were B19 DNA positive in cardiac tissue (18.92%), while in control group, including 8 RHD patients (autopsy specimens) and 20 non-CHD patients, the B19 gene were all negative, there was a significant difference of B19 gene presence between CHD group and control group (P = 0.0421). ISH of B19 gene in cardiac tissue were positive in 4 CHD patients and all were negative in control group. The B19 gene was found locating in the nucleus of cardiac cell by ISH. CONCLUSIONS: Parvovirus B19 infection correlated with CHD. Since B19 gene was mainly found in the cardiac cell nucleus, maybe it influenced the regulation of gene expression and affected the development of heart with the result of CHD. PMID- 11503057 TI - [Seroepidemiology of adult diarrhea rotavirus in some countries of Asia]. AB - OBJECTIVE: Adult diarrhea rotavirus (ADRV) is the major cause of severe adult diarrhea. This paper is to survey the ADRV antibody in sera of normal persons of some areas of Asia including China cities of Changchun, Nanking, Taiyuan, Wuhan, Kathmandu of Nepal, Seoul of South Korea and Hong Kong China. METHODS: The ADRV antibody was detected by enzyme-linked immunosorbent blocking test. RESULTS: The data of survey showed that 1. A total detection of 2183 normal sera from persons of Asian areas showed an ADRV antibody positive rate of 3.6% (79/2183); 2. In cities in China and in Hong Kong, the ADRV antibody showed a lower level in age groups of 10-20,20-30-; 3. The ADRV antibody was detectable in normal persons in 1997 in Hong Kong which indicated an earlier existence of ADRV there before the ADRV outbreak in mainland China in 1983; 4. The ADRV antibody level observed in 1986-1991 in Hong Kong showed no obvious difference; 5. ADRV antibody was detectable in normal persons in Nepal and South Korea. CONCLUSIONS: The survey showed there was only a low level of ADRV antibody in normal persons in the Asian areas surveyed. PMID- 11503058 TI - [The arboviruses of genus alphavirus family togaviridae were isolated from mosquitoes captured from Yantai]. AB - OBJECTIVE: To isolate arbovirus from mosquitoes caught from Yantai. METHODS: The isolated viruses were tested for their physico- chemical properties, examined by electron microscopy and specific immuno-reactivity. RESULTS: Fifteen strains of virus were isolated from mosquitoes in 1994 from Yantai. Three of them were further assayed. The results showed that the viruses could multiply on C6/36 cell and produce typical cytopathogenic effect. The viruses couldn't cause regular sickness and death of suckling mice by intracerebral inoculation. The viruses were sensitive to ether, but resistant to 5, -Idu. One sample was examined with electron microscope, and spherical virus particles were observed. The diameter of the virus particles is about 55 +/- 2. 3nm. It did not react with the JBE virus and Bunyaviridae group specific immuno-ascitic fluids but cross-reacted with the group A Togaviridae specific immuno-ascitic fluid. CONCLUSIONS: The results showed that the viruses isolated from Yantai belong to alphavirus of togaviridae. PMID- 11503059 TI - [Detection of Japanese encephalitis virus in samples of JE patients by RT-PCR]. AB - OBJECTIVE: To set up and assess the feasibility of RT-PCR method for Japanese encephalitis virus (JEV) detection, and use this method to detect clinical specimens. METHODS: The sensitivity of this RT-PCR was measured by plaque formation test, and the specificity of primers was proved by detecting some other flaviviruses. This RT-PCR was also compared with PRHI. RESULTS: JEV RNA could be detected successfully from viral culturing supernatants and from mouse brain infected by JEV. Its sensitivity of detecting JEV RNA from viral culturing supernatants was 64 PFU. A total of 38 specimens (of which 25 were serum specimens, 13 CSF specimens) from 16 clinically susceptible JE patients were examined by RT-PCR and RPHI Summing up the detection results of RT-PCR and RPHI, 14 patients among the 16 susceptible patients were affirmed as Japanese encephalitis patients. This RT-PCR is specific to JEV. The sensitivity of joint use of RT-PCR and RPHI to detect JEV was 100%, higher than using RPHI alone, and was 7.7% higher than using RT-PCR only. RT-PCR is more suitable for epidemiological survey than virus isolation. CONCLUSIONS: This RT-PCR for JEV detection is highly specific and sensitive. Toxemia period of JE patient is very short, so grasping the time of blood specimen collection will affect virus detection rate. PMID- 11503060 TI - Development of the fragment constant method for estimating the partition coefficients of nonionic organic mixtures. AB - The fragment constant model for estimating partition coefficient of nonionic organic mixtures is developed based on group contribution theory. For 60 mixtures randomly composed of eight substituted benzenes, the C(18)-Empore disks/water partition coefficients (K(MD)) are determined. Establishment of a model between these K(MD) and the mole fraction of the fragments (Cl, Br, I, NO(2), CH(3)) acquires the fragments constants for K(MD). The significant linear regression equation, between these fragment constants and that of Hansch, provides an approach for calculating K(MD) of untested nonionic organic mixtures by using the fragment constants of Hansch. PMID- 11503061 TI - Gridded usage inventories of technical hexachlorocyclohexane and lindane for China with 1/6 degrees latitude by 1/4 degrees longitude resolution. AB - China banned the use of technical HCH (BHC: 1,2,3,4,5,6-hexachlorocyclohexane) in 1983; lindane has been used in this country since 1991. The total production was around 4 million t for technical HCH from 1952 to 1984, and 11,400 t for lindane between 1991 and 2000. Though the total produced technical HCH was considered to be used in China, only 3,200 t of lindane were used between 1991 and 2000 with the rest for export or on unused stack. Annual usage of these two compounds was located into each province first and then broken for different crops. Inventories of gridded usage of both technical HCH and lindane with 1/6 degrees by 1/4 degrees latitude/longitude resolution have been created by different gridded cropland data sets as surrogate data. The intensive use of technical HCH on croplands was concentrated in the southeastern part of China, but the use of lindane was concentrated in the northern part of China. PMID- 11503062 TI - Aerobic and anaerobic metabolism of glutaraldehyde in a river water-sediment system. AB - Material balance studies of glutaraldehyde in a river water-sediment system demonstrate that glutaraldehyde preferred to remain in the water phase. Glutaraldehyde was metabolized rapidly under both aerobic and anaerobic conditions. The pseudo-first-order half-life of catabolism, based on the loss of glutaraldehyde from the water phase, was 10.6 h aerobically and 7.7 h anaerobically. In contrast, under sterile conditions at pH 5 or 7, no appreciable degradation of glutaraldehyde was observed over a 31-day period. At pH 9, about 30% of the glutaraldehyde degraded over the same period. The major degradate was identified as 3-formyl-6-hydroxy-2-cyclohexene-1-propanal, a cyclicized dimer of glutaraldehyde. The extrapolated half-life of abiotic degradation was 508 days at pH 5, 102 days at pH 7, and 46 days at pH 9. Under aerobic conditions, glutaraldehyde was first biotransformed into the intermediate glutaric acid, which then underwent further metabolism ultimately to carbon dioxide. Metabolism of glutaraldehyde under anaerobic conditions did not proceed ultimately to methane, but terminated with the formation of 1,5-pentanediol via 5 hydroxypentanal as an intermediate. PMID- 11503063 TI - Nitrogen, phosphorous, and organic carbon removal in simulated wetland treatment systems. AB - Effects of vegetation, wastewater drawdown, hydraulic retention time (HRT), and media depth on removal of nitrogen, phosphorous, and organic carbon in microcosms were investigated. Synthetic wastewater was added daily to 28 microcosms, and effluent was sampled every 12 days for 132 days. Effluent was analyzed for ammonium (NH(4)(+)-N), nitrate (NO(3)(-)-N), orthophosphate (PO(4)(3-)-P), and total organic carbon (TOC). Average percent removal of NH(4)(+)-N was significantly greater in microcosms containing plants (67%) than in those without plants (29%). Percent removal of PO(4)(3-)-P was also significantly greater in microcosms with plants (42%) than in microcosms without plants (20%), but no significant difference was found for TOC removal between microcosms with plants (67%) and those without plants (74%). Average removal was significantly lower in microcosms with wastewater drawdown than in those without wastewater fluctuation for both NH(4)(+)-N (51% versus 83%) and PO(4)(3-)-P (14% versus 71%). Percent NH(4)(+)-N removal was significantly greater in microcosms with a 6-day retention time (80%) than in those with a 2-day retention (53%), and PO(4)(3-)-P removal was also significantly greater with a 6-day retention time (55%) than a 2-day retention (29%). No differences were seen in TOC removal due to any of the treatments but HRT, where removal was greater microcosms with a 2-day HRT (76%) than in those with a 6-day HRT (60%). Media depth did not have a significant effect on nutrient removal. Results of this study demonstrate that required design parameters are different depending on the nutrient being removed in systems simulated by these microcosms. PMID- 11503064 TI - Effects of experimental manipulation of pH and salinity on Cd(2+) uptake by the sponge Microciona prolifera and on sponge cell aggregation induced by Ca(2+) and Cd(2+). AB - Marine sponges (Microciona prolifera) collected in St. Joseph Bay, Florida panhandle, were exposed for 2 h to pH/salinity unit combinations of 7.4/30, 6.3/30, 7.4/11, and 6.3/11. Cell suspensions from these were aggregated with 24 microM of either CaCl(2) or CdCl(2). Cells exposed to the low/low (11/6.3) combination aggregated spontaneously (no added stimulus) in 8/11 experiments, suggesting a significant disturbance of normal function, possibly involving disrupted ion uptake. In all other combinations aggregation proceeded normally and there were no statistically significant differences among the groups. CdCl(2) was as effective an aggregation stimulus as CaCl(2). The calcium channel blocker verapamil (100 microM) reduced calcium-induced aggregation by 15% but had no effect on cadmium (Cd)-induced aggregation, indicating that L-type calcium channels do not play a major role in aggregation induced by these divalent cations. Sponge tissue was exposed for 48 h to the same pH/salinity combinations but containing Cd (15 or 150 microg/ml) and then dried and analyzed for Cd. All sponges concentrated Cd but those exposed to low salinity concentrated it most (in one case x13). Low pH alone had no appreciable effect but appeared to increase the effect of low salinity. One sponge with a native Cd content of 47.2 microg/g dry weight had the highest acquired Cd content. The results of this study indicate that low levels of salinity and pH, similar to those we recorded in the study area, facilitate the accumulation of Cd, but not via L-type calcium channels, and disrupt normal aggregation responses of the cell. These results may help explain a previous observation that cells from M. prolifera from this area, with high native levels of Cd, failed to aggregate in response to CaCl(2)[Philp RB (1999) Comp Biochem Physiol 124C:41-49] and also the frequent die-offs of Microciona that have virtually eliminated this sponge from its local habitat. PMID- 11503065 TI - Chemical contamination and toxicity of sediment from a coastal area receiving industrial effluents in Kuwait. AB - The Shuaiba coastal area (12.5 x 1.5 km) was examined for contamination with total organic carbon, volatile organic matter, total petroleum hydrocarbons, polycyclic aromatic hydrocarbons, cadmium, chromium, copper, lead, nickel, vanadium, and zinc in sediment; their desorption by aqueous elution; and toxicity to aquatic biota. The pollutants were mainly accumulated in the upstream area facing Mina Al-Ahmadi refinery to Shuaiba harbour. Solid-phase Microtox assays showed severe toxicity, and the LC(50) was negatively correlated with most of the chemical parameters, suggesting that toxicity was the function of collective effects of the pollutants present in sediment. Sea water elutriation showed poor desorption of pollutants from sediment, and the elutriates were not found toxic to Microtox and brine shrimp larvae. Whole sediment suspension in sea water reduced the survival of fingerlings in fish bioassays. Action from Shuaiba Area Authority is required to reduce pollutant accumulation in identified depositional area on the Shuaiba coast by facilitating unrestricted water flow in the area and restricting pollutant discharge at source. PMID- 11503066 TI - Sediment toxicity identification evaluation (TIE) studies at marine sites suspected of ordnance contamination. AB - A sediment quality assessment survey and subsequent toxicity identification evaluation (TIE) study was conducted at several sites in Puget Sound, Washington. The sites were previously suspected of contamination with ordnance compounds. The initial survey employed sea urchin porewater toxicity tests to locate the most toxic stations. Sediments from the most toxic stations were selected for comprehensive chemical analyses. Based on the combined information from the toxicity and chemical data, three adjacent stations in Ostrich Bay were selected for the TIE study. The results of the phase I TIE suggested that organics and metals were primarily responsible for the observed toxicity in the sea urchin fertilization test. In addition to these contaminants, ammonia was also contributing to the toxicity for the sea urchin embryological development test. The phase II TIE study isolated the majority of the toxicity in the fraction containing nonpolar organics with high log K(ow), but chemical analyses failed to identify a compound present at a concentration high enough to be responsible for the observed toxicity. The data suggest that some organic or organometallic contaminant(s) that were not included in the comprehensive suite of chemical analyses caused the observed toxicological responses. PMID- 11503067 TI - Development of marine toxicity data for ordnance compounds. AB - A toxicity database for ordnance compounds was generated using eight compounds of concern and marine toxicity tests with five species from different phyla. Toxicity tests and endpoints included fertilization success and embryological development with the sea urchin Arbacia punctulata; zoospore germination, germling length, and cell number with the green macroalga Ulva fasciata; survival and reproductive success of the polychaete Dinophilus gyrociliatus; larvae hatching and survival with the redfish Sciaenops ocellatus; and survival of juveniles of the opossum shrimp Americamysis bahia (formerly Mysidopsis bahia). The studied ordnance compounds were 2,4- and 2,6-dinitrotoluene, 2,4,6 trinitrotoluene, 1,3-dinitrobenzene, 1,3,5-trinitrobenzene, 2,4,6 trinitrophenylmethylnitramine (tetryl), 2,4,6-trinitrophenol (picric acid), and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). The most sensitive toxicity test endpoints overall were the macroalga zoospore germination and the polychaete reproduction tests. The most toxic ordnance compounds overall were tetryl and 1,3,5-trinitrobenzene. These were also the most degradable compounds, often being reduced to very low or below-detection levels at the end of the test exposure. Among the dinitro- and trinitrotoluenes and benzenes, toxicity tended to increase with the level of nitrogenation. Picric acid and RDX were the least toxic chemicals tested overall. PMID- 11503068 TI - A bioassay for the measurement of insecticide concentration. AB - A bioassay was developed to measure insecticide residues using fruit flies (Drosophila melongaster). After adding a known volume of sampling solution, the time at which 50% of the flies were dead (LT(50)) was recorded and cross referenced to the appropriate calibration curve. Using known standards, comparable results were obtained using the bioassay and GC-MS. The bioassay allows concentrations of synthetic pyrethroids as low as 1 pg L(-1) to be measured with a variance of < 5%. The bioassay can be used reliably over a wide range of temperatures and it is tolerant to a range of pH and surface tensions of the test solution. The whole bioassay is compact, physically robust, and simple to use; hence, it could be of use in the field as a quick preliminary assessment of water contamination. PMID- 11503069 TI - Toxicity of fipronil and its degradation products to Procambarus sp.: field and laboratory studies. AB - Fipronil is a phenylpyrazole insecticide that is the active ingredient in the pesticide Icon 6.2 FS which is applied to rice seeds targeting the rice water weevil. An arthropod-selective insecticide, fipronil blocks the GABA-gated chloride channel and is unique in that several of its degradation products have been indicated to be equal or more potent than fipronil. After application of rice seeds (2-3 days postplant) to flooded rice fields, water is typically pumped from the rice fields and can be used for the culture of crayfish (Procambarus sp.). Because fipronil is selective for arthropods, is transported via organic sediment, and crayfish consume organic sediment, 96-h LC(50) experiments were conducted with fipronil and three of its environmental derivatives in crayfish under conditions without carrier solvents in water of similar pH, alkalinity, and hardness as observed in south Louisiana crayfish culture ponds. Measured LC(50)s for fipronil to red swamp (Procambarus clarkii) and white river (Procambarus zonangulus) crayfish were 14.3 (95% CI; 5.1-23.4) and 19.5 (95% CI; 11.1-27.9) microg/L, respectively. LC(50)s of fipronil sulfone (11.2; 9.2-13.2 microg/L), fipronil sulfide (15.5; 13-18 microg/L); and the photoproduct, desulfinyl fipronil (68.6; 46-95.2 microg/L) displayed very high toxicity in crayfish. In situ toxicity studies using caged crayfish in culturing ponds receiving effluent from drained rice fields indicated that effluent from rice fields planted with Icon-treated seed was significantly more toxic compared to untreated surface water (40% survival compared to 83% survival). Hazard quotient comparisons using measured water concentrations in the field and laboratory-based LC(50)s indicated that fipronil and its metabolites in water resulting from Icon-treated rice seed planting poses a significant risk to crayfish survival. PMID- 11503070 TI - Toxicity of cadmium and copper on larval and juvenile stages of the estuarine crab Chasmagnathus granulata (Brachyura, Grapsidae). AB - The toxicity of cadmium and copper were assayed in the estuarine crab Chasmagnathus granulata. Acute assays were made on the first larvae stage, and both acute and chronic assays were made on juvenile crabs. The acute lethal toxicity of the assayed heavy metals was three orders of magnitude higher in larvae than in juveniles. Cadmium proved to be more toxic than copper in most cases; this difference was more evident in the chronic assays on juveniles, according to the nonphysiological feature of cadmium and its persistent accumulation in organisms. During these chronic assays, cadmium produced both a significant mortality and a clear retardation of molting, in accordance with its inhibitory effect on the molting process as previously reported. Copper only caused a molting acceleration during chronic assays, perhaps as a detoxifying mechanism. Heavy metal concentrations having effects in the chronic assays have been reported in some sectors of the estuary where the assayed species lives. PMID- 11503071 TI - Analysis and occurrence of polybrominated diphenyl ethers in Washington state freshwater fish. AB - A method is described for analyzing polybrominated diphenyl ethers (PBDEs) in fish tissue using gas chromatography with atomic emission detection (GC/AED) and quantitation by compound independent calibration (CIC). The method has the advantage of an element-selective response, including selectivity between bromine and chlorine. An analysis of fish tissue samples from selected locations in Washington State showed total PBDE concentrations ranged from 1.4 microg/kg (wet weight) in rainbow trout from a remote spring-fed stream to 1,250 microg/kg in mountain whitefish from the urbanized Spokane River. Tetra and penta isomers were the major compounds present. There appear to be substantial between-species differences in the ability of fish to metabolize PBDEs. PMID- 11503072 TI - Effect of endosulfan on antioxidants of freshwater fish Channa punctatus Bloch: 1. Protection against lipid peroxidation in liver by copper preexposure. AB - Effect of a single exposure of endosulfan (5 ppb) on catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), and reduced glutathione (GSH) of liver, kidney, and gill of a freshwater fish (Channa punctatus Bloch) were evaluated after 24 h of treatment. Endosulfan exposure resulted in a significant induction (p < 0.05-0.001) of GPx, GST activity, and GSH levels in all the organs. However, CAT activity was found to be significantly decreased (p < 0.01 0.001). Lipid peroxidation (LPO) values were also determined in liver, kidney, and gill and a significant increase in LPO values (p < 0.05-0.01) was observed in all the organs. We also investigated whether preexposure to low concentration of copper (10 ppb) for 4 weeks has any protective effect against endosulfan-induced oxidative damage. In copper-acclimatized endosulfan-exposed fish, a significant decrease in GPx (p < 0.001), GST (p < 0.05), GSH (p < 0.001) levels, and LPO (p < 0.01) was observed in the liver, whereas CAT activity was increased significantly (p < 0.001). However, kidney and gill did not show any significant alterations in antioxidant levels. The results of this study demonstrate that endosulfan induces peroxidative damage in liver, kidney, and gill in response to which levels of antioxidant were modulated. However, when fish preacclimatized to copper were exposed to endosulfan, protection against oxidative damage was observed only in the liver. It is proposed that measurement of antioxidants in fish tissues may prove to be useful in biomonitoring of exposure to aquatic pollutants. PMID- 11503073 TI - Accumulation profiles of persistent organochlorines in waterbirds from an estuary in Korea. AB - Persistent organochlorine pollutants (POPs), such as polychlorinated dibenzo-p dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), and organochlorine (OC) pesticides such as DDTs (dichlorodiphenyltrichloroethane), HCHs (hexachlorocyclohexane isomers), CHLs (chlordane compounds) and HCB (hexachlorobenzene), were measured in subcutaneous fat of resident and migratory birds collected from the Nakdong River estuary (NRE) in Korea. Black-tailed gull, a resident bird from the NRE, contained greater concentrations of PCDD/Fs and PCBs than the migratory birds collected in the estuary. For example, mean concentrations of PCDD/Fs in black-tailed gull (395.5 pg/g fat weight) were higher than those in migratory birds, such as greenshank (198.3 pg/g fat weight), common gull (90.9 pg/g fat weight) black headed gull (84.2 pg/g fat weight), and common tern (47.1 pg/g fat weight). However, concentrations of DDTs and/or HCHs were great in some migratory species, such as little tern (mean DDT 6,200 ng/g fat weight) and black-necked grebe (HCHs 475 ng/g fat weight). This suggested that contamination of PCDD/Fs and PCBs in resident gulls are due to intake of locally contaminated fish near the NRE. Elevated OC pesticide levels in migratory birds indicated that these birds have been exposed to DDTs and HCHs during their migration in Southeast Asian countries where chlorinated pesticides are still used. 2,3,7,8-Tetrachloro dibenzo-p-dioxin equivalents (TEQs) were calculated using the toxic equivalency factors (TEFs) reported by World Health Organization in 1998. Four of the 2,3,7,8-substituted congeners of PCDD/Fs contributed over 90% of the TEQs. PMID- 11503074 TI - Lead poisoning of waterfowl by contaminated sediment in the Coeur d'Alene River. AB - The Coeur d'Alene River basin in Idaho has been contaminated by mine tailings that have impaired the health of wildlife since the early 1900s. In other parts of the world, virtually all lead poisoning of waterfowl is caused by the ingestion of manmade lead artifacts, primarily spent lead shotshell pellets or, occasionally, fishing sinkers. However, in the Coeur d'Alene River basin in Idaho, nonartifactual lead poisoning was the ultimate cause of death of most of 219 (77%) of 285 waterfowl carcasses that had been found sick or dead from 1992 through 1997. The majority of these 219 waterfowl (172 tundra swans [Cygnus columbianus], 33 Canada geese [Branta canadensis], and 14 other species) were poisoned by ingesting river sediment that was contaminated with lead. The next most common cause of death (20 instances, 7%) was lead poisoning accompanied by ingested shotshell pellets. The remaining 46 waterfowl succumbed to trauma, infectious diseases (aspergillosis, avian cholera, tuberculosis), or miscellaneous problems, or the cause of death was not determined. PMID- 11503075 TI - Secondary effects of the pesticide Bacillus thuringiensis kurstaki on chicks of spruce grouse (Dendragapus canadensis). AB - The objective of this study was to determine if application of the biological pesticide Bacillus thuringiensis kurstaki (B.t.k.) had secondary effects on spruce grouse (Dendragapus canadensis) chicks in the southern boreal forest of Ontario, Canada. B.t.k. kills larvae of Lepidoptera that might otherwise be food. This may affect types of food ingested and influence chick growth rates by reducing the availability of a high-protein food. Amount of ingested grit, which is diet-dependent, may also be influenced, which could alter exposure to anthropogenic toxic metals, such as cadmium (Cd) and lead (Pb), associated with the grit. Chick growth rate from 0 to 14 days of age was determined, and gizzard contents were analyzed for types of foods and levels of macronutrients and trace metals in chicks raised in a plot treated with B.t.k. compared to chicks raised on a control plot. Chick growth rates were significantly different (p < 0.05); chicks from the treatment plot increased body mass 30% less as compared to chicks raised on the control plot. Control chicks contained on average 2.5-fold the mass (in g) of grit and nearly twofold the mass of food, which included larvae, in their gizzards compared to those exposed to B.t.k. whose diets were dominated by spiders and ants. Concentrations of magnesium (Mg), calcium (Ca), phosphorous (P), iron (Fe), manganese (Mn), sulfur (S) and the trace metals zinc (Zn), copper (Cu), nickel (Ni), and chromium (Cr) did not differ between experimental and control birds (p > 0.05). In contrast, control chicks had higher levels of aluminum (Al), Cd, cobalt (Co), and Pb in their gizzards than chicks from the sprayed area (p < 0.05), attributed to greater amounts of consumed grit. Although birds from the sprayed area had lower levels of the toxic metals Pb and Cd in their gizzards, a protein-deficient diet could have been a factor that contributed to decreased rates of growth in B.t.k.-exposed birds as compared to those birds held on nonsprayed sites. Reduced growth during the first 2 weeks of avian development has important consequences at the population level, and it is recommended that this secondary effect be considered for all species who rely on Lepidoptera larvae as a food resource prior to wide spread application of B.t.k. for insect control. PMID- 11503076 TI - Effects of mercury on the contractile activity of the right ventricular myocardium. AB - The increase in right ventricular systolic pressure observed in vivo after the administration of mercury opposes to the idea that the metal depresses the cardiac pump performance. We then investigated the effects of HgCl(2) (0.1 to 2.5 microM) on the contractile activity of the right ventricular myocardium, measuring isometric and tetanic contractions of right ventricular isolated strips, right ventricular isovolumic systolic and diastolic pressures, and the coronary perfusion pressure (0.03 to 3 microM) in constant-flow Langendorff perfused rat hearts. The results presented here suggest that the acute effects of mercury on the right ventricular myocardium are distinct. When isolated strips of right ventricular wall are used, the contractile depression produced by mercury is manifested. However, when mercury is administered to isolated perfused hearts or in vivo this depressant effect is not revealed. The possible reasons for this behavior are the increased coronary perfusion pressure, which promotes a positive inotropic effect, manifested during the infusion of increasing concentrations of mercury, or the putative stretch of the ventricular fibers, which might cause the increment of diastolic pressure. An interesting finding is that the mechanical activity of the preparations, in which mercury is administered via coronary circulation, is not depressed and, even more, it can increase systolic pressure. However, the nature of this protective effect of coronary circulation cannot be explained by the results presented here. PMID- 11503077 TI - Hepatic enzyme induction and acute endocrine effects of 2,2',3,3',4,6' hexachlorobiphenyl and 2,2',3,4',5',6-hexachlorobiphenyl in prepubertal female rats. AB - Polychlorinated biphenyls (PCBs) with the liable 2,3,6-substitution are important components of certain commercial mixtures and frequently detected in biota, but little is known about their enzyme induction abilities and possible endocrine disrupting effects. CB 132 (2,2',3,3',4,6'-hexachlorophenyl) and CB 149 (2,2'3,4',5',6-hexachlorophenyl) were investigated in weanling female rats dosed intraperitoneally on days 21 and 22 and killed on day 24 of age. Uterotropic response, serum thyroid hormone, and hepatic enzyme induction were examined in prepubertal female rats treated with these two environmentally relevant 2,3,6 substituted chlorobiphenyl (CB) congeners from 8 mg/kg to 96 mg/kg. The readily metabolized CB 132 did not cause any significant increase in all endpoints measured in the present study. On the other hand, CB 149 was a weak PROD and BROD inducer and a modest depleter of serum thyroxine in prepubertal female rats. The finding of thyroid hormone disruption by CB 149 may lead to biologically significant neurobehavioral and neurochemical changes in developing animals via milk lactation. PMID- 11503078 TI - Organochlorines and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in cancerous and noncancerous breast tissue: do the data support the hypothesis that oxidative DNA damage caused by organochlorines affects breast cancer? AB - A study was conducted to test the hypothesis that oxidative DNA damage caused by exposure to organochlorines is an important risk factor in breast cancer. This is the first study that evaluates this hypothesis by measuring 8-hydroxy-2' deoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage, polychlorinated biphenyl (PCB) congeners, and isomers of bis (4-chlorophenyl)-1,1,1 trichloroethane (DDT) and bis (4-chlorophenyl)-2,2,2-dichloroethane (DDE) in cancerous and noncancerous tissue. We measured these compounds in 44 primary tumors (cancerous) and 21 benign breast biopsy (noncancerous) tissues. Overall, no significant differences were observed in the level of the organochlorines between the tissues. The median concentration for 8-OHdG was 10.5 fmol/mg DNA (1.7/10(5) deoxyguanosine residues), and 8.5 fmol/mg DNA (1.4/10(5) deoxyguanosine residues) in cancerous and noncancerous tissue, respectively. These values are similar to background levels. No significant differences were observed in 8-OHdG levels in cancerous versus noncancerous tissue, and no correlation was demonstrated between the organochlorines and 8-OHdG. The data thus do not support the hypothesis that oxidative DNA damage caused by exposure to organochlorines is an important risk factor in breast cancer. PMID- 11503079 TI - Abdominal aortic stenting: current practice. PMID- 11503080 TI - Endoluminal treatment of abdominal aortic aneurysms. AB - BACKGROUND: We report our preliminary results with endovascular treatment of abdominal aortic aneurysms (AAA). METHODS: Between October 1998 and June 2000, 64 patients (62 male, two female; mean age = 70 years) underwent endovascular repair of AAA. Different types of prostheses were used, both bifurcated (n = 58) and straight (n = 6). We performed duplex sonography and spiral computed tomographic angiography (CTA) at discharge and at 3, 6, 12, and 18 months. Follow-up ranged from 1 to 20 months. RESULTS: All procedures were successful, except for three immediate and one late surgical conversions (6.2%). One patient died 14 days after immediate surgical conversion. At discharge, CTA showed 13 endoleaks: three resolved spontaneously, six persisted during follow-up, and four (one angioplasty and three embolizations) were treated successfully. Stenosis of an iliac branch occurred in one patient after 3 months and was successfully treated by angioplasty. Late endoleaks were detected by imaging follow-up in four cases, three at 1 year and one at 6 months, requiring deployment of distal extender cuffs (n = 2), a proximal cuff (n = 1), and lumbar embolization (n = 2). CONCLUSION: Our preliminary experience supports the efficacy of endovascular repair in selected patients, but strict and accurate follow-up is required. PMID- 11503081 TI - Duodenal carcinoma: sonographic findings. AB - BACKGROUND: Duodenal carcinoma is a relatively rare tumor and its sonographic (US) findings have rarely been reported. METHODS: We reviewed the clinical data and US results of 15 patients with histologically proven duodenal carcinoma to pinpoint the contribution of US to the diagnosis. RESULTS: US was useful for not only detecting the duodenal carcinoma (13 of 15, 86.7%), which altered the endoscopic procedure in nine patients, but also for evaluating vascular involvement (marked portal invasion in four cases and inferior vena cava invasion in two cases). Four of the eight carcinomas arising from the second or third portion showed this vascular invasion. CONCLUSION: After reviewing the clinical data and US results of 15 cases of duodenal carcinoma, we found that US was useful for detecting duodenal carcinoma and determining vascular invasion. Thus, US contributed significantly to alter the endoscopic procedure and patient management. PMID- 11503082 TI - Primary small bowel tumors: a radiologic-pathologic correlation. AB - BACKGROUND: Primary small bowel tumors are rare and their preoperative diagnosis is unsatisfactory. The cornerstone of diagnosis remains contrast radiography. The present study was done to evaluate the radiologic findings of primary small bowel tumors as shown on enteroclysis and to correlate these observations with surgical and histopathologic findings. METHODS: Seventy two patients with primary small bowel tumors identified by enteroclysis were evaluated. All the patients were subjected to jejunal biopsy or surgery. The diagnosis was established by histopathologic examination in all cases. RESULTS: Radiologic findings were suggestive of benign tumors in 19 patients and malignant tumors in 31 patients. Nonspecific findings in the form of diffuse involvement of the small bowel were noted in 22 patients. There was 100% radiologic-surgical correlation. Leiomyomas and lymphomas were the most common benign and malignant tumors, respectively. Tumor specificities were 89.5% for benign tumors and 41.5% for malignant tumors. CONCLUSION: Distinctive morphologic patterns as shown on enteroclysis are highly suggestive of specific tumor types in the majority of cases. PMID- 11503083 TI - Enteroclysis: a multidisciplinary approach. AB - We developed a method for passing the enteroclysis catheter at endoscopy in patients requiring upper endoscopy and enteroclysis. This method reduced patient discomfort, the time needed for fluoroscopic tube placement, and overall radiation doses. We conclude that endoscopy and enteroclysis performed together is practical in these patients. PMID- 11503084 TI - Sonographic diagnosis of adult intussusception. AB - We describe three cases of adult intussusception in which ultrasonography provided the correct preoperative diagnosis. Patients underwent ultrasonography to investigate nonspecific acute abdominal pain; intussusception was not suspected. In all cases, the sonographic pattern was typical of intussusception and ultrasonography was the only diagnostic study. Bowel ischemia was not found at surgery in any patient. PMID- 11503085 TI - Liver tumors in fatty liver: difficulty in ultrasonographic interpretation. AB - BACKGROUND: Fatty liver and liver tumors are very frequent diseases. Sonography (US) currently is the initial diagnostic tool for hepatic exploration. However, there is a marked paucity of US findings of tumors in fatty liver. METHODS: We studied the US findings of 41 lesions (31 patients) with this combination, with special attention paid to internal echoes, marginal echoes, and especially the mode of back echoes, and compared them with the US results of 64 lesions (38 patients) without fatty liver. RESULTS: Comparing the group having liver tumor without fatty liver with the group having liver tumor with fatty liver showed that (a) the number of hypoechoic lesions increased (29 of 41, 70.7%, vs. 16 of 64, 25.0.%), (2) the tumor margin appeared indistinct (32 of 41, 78.0%, vs. nine of 64, 14.1%), and (c) the lesions showing posterior echo enhancement increased (34 of 41, 82.9%, vs. eight of 64, 12.5%). CONCLUSION: Liver tumors in fatty liver are expected to show unusual patterns on US, so we should consider this difficulty when interpreting these US findings and we should not make a conclusion without including other imaging modalities. PMID- 11503086 TI - Pericaval fat collection mimicking an intracaval lesion on CT in patients with chronic liver disease. AB - BACKGROUND: Localized fat collections, which mimic fat-containing lesions of the inferior vena cava (IVC) on computed tomographic (CT) images, have been reported as rare incidental findings. Our goal was to evaluate the association of this CT finding with chronic liver disease. METHODS: Sixty-one patients with chronic liver disease were prospectively studied with contrast-enhanced abdominal CT. The prevalence, attenuation, location, shape, and size of the pericaval fat collections were assessed. Multidirectional reformatted CT images were obtained from helical CT data to identify origins of the pericaval fat collections. Sixty one patients without chronic liver disease were studied as control subjects. RESULTS: Pericaval fat collections were seen on abdominal CT in 16 (26.2%) of the 61 patients. On the reformatted images, the fat collections were contiguous to the fat tissue around the subdiaphragmatic esophagus in all 16 patients. The fat collections were located at the posterior aspect of the IVC in 12 patients. In the control group, pericaval fat collection mimicking an intracaval lesion was not seen. CONCLUSION: In patients with chronic liver disease, pericaval fat collections are not rare CT findings and their characteristic location is considered to be posterior to the IVC. It is important not to misinterpret such CT findings as abnormalities of the IVC, such as thrombus or tumors. PMID- 11503087 TI - Sclerosed hemangioma of the liver. AB - We report the radiologic findings of sclerosed hemangioma (SH), a rare variant of hepatic hemangioma. Dynamic contrast-enhanced computed tomography showed a hypodense mass in the liver with delayed enhancement. T2-weighted magnetic resonance imaging showed the mass as hypointense in relation to cerebrospinal fluid. The final diagnosis of SH was made pathologically. Although SH is rare, understanding its radiologic appearance is important to avoid unnecessary surgery and should be included in the differential diagnoses of hepatic lesion with delayed enhancement. PMID- 11503088 TI - Sonographic features of hepatic adenomas with pathologic correlation. AB - BACKGROUND: We compared the sonographic characteristics of hepatic adenomas with pathologic findings. METHODS: Information over 10 years was collected on 12 patients (six men, six women; mean age = 47 years) with surgically proven hepatic adenomas. Clinical data, sonographic features, and histopathologic findings were reviewed. RESULTS: The tumors in males were smaller and simpler than those in women (p < 0.05, Fisher's exact test). Four of the six larger tumors (>5 cm) showed mixed-echoic patterns corresponding with pathologically intratumoral hemorrhage and necrosis. Four homogeneously hypoechoic tumors had less change in tumor composition. Three homogeneously hyperechoic tumors had evident fatty changes inside. One isoechoic tumor had a hypoechoic rim, that correlated mostly to the tumor itself and compressed liver parenchyma. Seven of the 12 tumors had thin fibrous capsules that were not seen on sonography. CONCLUSION: Hepatic adenomas have variable sonographic appearances depending on changes in the tumor. Hypoechoic, hyperechoic, and mixed-echoic patterns represent simple adenoma, adenoma with fatty metamorphosis, and hemorrhagic necrosis, respectively, in tumors. PMID- 11503089 TI - Peliosis hepatis: triphasic helical CT and dynamic MRI findings. AB - We report the triphasic helical computed tomographic (CT) and dynamic magnetic resonance imaging (MRI) findings in a unique case of biopsy-proven peliosis of the liver. Several reports have described the CT and MRI findings of this entity without specific appearance. This report discusses the usefulness of dynamic helical CT and MRI for the early diagnosis of peliosis. PMID- 11503090 TI - Transcatheter arterial embolization in the treatment of symptomatic cavernous hemangiomas of the liver: a prospective study. AB - BACKGROUND: This prospective study evaluated the clinical and radiologic results of transcatheter arterial embolization (TAE) for the treatment of symptomatic cavernous hemangiomas of the liver. The technique, its complications, and effectiveness also were analyzed. METHODS: Eight patients (five male, three female; mean age +/- SD = 47.75 +/- 8.59 years) with symptomatic cavernous hemangiomas of the liver were treated by TAE with polyvinyl alcohol particles or gelfoam and steel coils (single session) followed by supportive treatment. Tumor characterization (including the extent and number of lesions) was done on triple phase helical computed tomography or gadolinium-enhanced dynamic magnetic resonance imaging. RESULTS: The lesions were located in the right lobe in five patients, left lobe in one, and both lobes in two. The largest diameter of the lesions was 6-18 cm (9.28 +/- 5.13 cm). The treatment response was assessed on follow-up ultrasound and color Doppler and/or contrast-enhanced helical computed tomography. There were no treatment-related deaths and morbidity was minimal. Embolization was the only method of treatment in seven patients; however, one patient had surgery after TAE because the symptoms were only partly relieved. Indications for embolization were abdominal pain (eight patients), rapid tumor enlargement (four of eight), and recurrent jaundice (one of eight). Symptomatic improvement was documented in all patients after embolization. Symptoms did not worsen in any patient. The mean size of the tumor did not show any statistically significant change on follow-up radiologic examinations. However, in one patient, the tumor significantly regressed in size after embolization. CONCLUSION: TAE of hepatic cavernous hemangioma is a useful procedure in the therapy of symptomatic hemangiomas. PMID- 11503091 TI - Hepatic falciform artery: angiographic observations and significance. AB - BACKGROUND: The purpose of this study was to assess the angiographic incidence and appearance of the hepatic falciform artery (HFA) and discuss its clinical significance. METHODS: Hepatic angiograms of 53 patients obtained with digital subtraction angiography were prospectively evaluated with regard to incidence, anatomic features, and flow speed of the HFA. We analyzed whether the background of chronic liver disease affected the incidence of the HFA. Transcatheter arterial chemoinfusion or chemoembolization for liver tumors was performed in 33 patients. We noted the occurrence of supraumbilical skin complications. RESULTS: The HFA was observed in 13 (24.5%) of 53 patients on celiac or common hepatic angiograms. The blood flow of the HFA was slower than that of the peripheral hepatic arteries in all patients. No significant difference in the incidence of HFA between the 34 patients with chronic liver disease and the 19 patients with normal livers was found. One treated patient with an HFA and a history of gastrectomy developed a supraumbilical red skin rash. CONCLUSION: The angiographic incidence of the HFA is more common than previously reported. The delayed and persistent opacification of the HFA on hepatic angiograms caused by its slow blood flow is considered the key to its identification. PMID- 11503092 TI - Intrahepatic portal vein aneurysm: three-dimensional power Doppler demonstration in four cases. AB - We describe four cases of portal vein aneurysm that were studied with ultrasonography, color Doppler, computed tomography, and magnetic resonance imaging as well as three-dimensional power Doppler. Three-dimensional ultrasonography showed the portal aneurysm and its relationship to its portal branches in all patients and showed the portosystemic communication in three. In all cases, computed tomography or magnetic resonance imaging confirmed the three dimensional ultrasonographic findings. PMID- 11503093 TI - Peripheral bone metastasis from hepatocellular carcinoma: sonographic findings. AB - BACKGROUND AND METHODS: We reviewed medical and radiologic records of five patients with bone metastasis from hepatocellular carcinoma (HCC) to evaluate the role of sonography (US) in the diagnosis of this bone metastasis. RESULTS: The metastatic lesions were clearly visualized by US as expansive, homogeneous, soft tissue masses with bony destruction, and color Doppler US showed fine vessels within the lesions. After treatment, US showed a decrease in tumor size with an increase in echogenicity and a decrease in blood flow. Hepatic US at the time of bone metastasis showed a portal tumor thrombus in all cases. CONCLUSION: When US detects a portal venous tumor thrombus in HCC patients, attention should be directed not only to the liver but also to bone to improve patient care. US is useful not only in detecting the metastatic lesion but also in evaluating the treatment effect. PMID- 11503094 TI - Splenic lymphoma: differentiation from splenic cyst with ultrasonography. AB - BACKGROUND AND METHODS: Lymphoma can be nearly anechoic and mimic a cyst on ultrasonography (US). To investigate whether this phenomenon occurs at the level of the spleen, we analyzed the US findings of 38 cases of splenic lymphoma and 16 cases of splenic cyst. RESULTS: (1) With regard to shape, echogenicity of the lesion, and mode of posterior echo, there was no difference between splenic lymphomas and splenic cysts. However, the boundaries of the lesions were indistinct in splenic lymphomas and distinct in splenic cysts. (2) Blood flow signals and vascular penetration were seen exclusively in splenic lymphomas. CONCLUSION: The mode of boundary echo (distinct or indistinct) distinguishes splenic lymphomas from splenic cysts. Color Doppler US increases the diagnostic confidence of US. PMID- 11503095 TI - Iliopsoas abscess: a report of 24 patients diagnosed by CT. AB - BACKGROUND: We wanted to define the role of computed tomography (CT) in the diagnosis, etiology, and treatment of iliopsoas abscess. METHODS: Twenty-four patients (18 men, six women; age range = 17-86 years) with iliopsoas abscesses diagnosed over 8 years were retrospectively reviewed. All presented with fever and elevated white blood cell counts. Twenty-one had abdominal, flank or pelvic pain and nine had specific psoas signs suggesting the diagnosis. RESULTS: Seventeen of the abscesses were right-sided. Twenty were regarded as secondary to various underlying causes that were clearly demonstrated on CT and related to gastrointestinal (n = 12), skeletal (n = 5), or urinary tract (n = 3) diseases. All patients received appropriate antibiotic treatment. Thirteen also had their abscesses drained and eight had definitive surgical procedures. CONCLUSION: CT is an effective imaging technique for diagnosing iliopsoas abscess, even when classic clinical signs are absent. Treatment by percutaneous drainage under CT guidance is another advantage. When a psoas abscess is a complication of Crohn's disease, resection of the affected bowel segment is recommended in addition to drainage because drainage alone even in conjunction with appropriate medical therapy is usually not effective. PMID- 11503096 TI - Transvaginal ultrasonography of nongynecologic pelvic lesions. AB - Transvaginal ultrasonography (TVUS) is one of the preferred imaging modalities in patients with gynecologic problems because of its high diagnostic accuracy, noninvasiveness, and wide availability. In endovaginal scanning, the problem of sonic attenuation is much less significant than with the transabdominal approach in the evaluation of the viscera in the true pelvis. Placement of high-frequency, high-resolution probes within the vagina allows accurate assessment of all anatomic structures of the female reproductive tract within the pelvis, and, incidentally, a variety of pathologic conditions affecting the intestinal tract, the urinary system, the pelvic walls, vessels, lymph nodes, and peritoneum can be assessed by this technique. In this article, we show the appearances of nongynecologic lesions of the female pelvis as imaged with TVUS and discuss the clinical indications to this kind of study and the role of TVUS in guiding interventional maneuvers through the vaginal vault. All endovaginal scans were taken with transducers at frequencies of 5.0-7.5 MHz. PMID- 11503097 TI - Juvenile granulosa cell tumor. AB - We present three-dimensional computed tomographic findings of a juvenile granulosa cell tumor of the ovary at FIGO stage IA in a 17-year-old woman. Juvenile granulosa cell tumor is one of the rare sex cord stromal tumors of the ovary. Most tumors at FIGO stage IA have a favorable prognosis, whereas those at higher stages have a less favorable outcome. PMID- 11503098 TI - Left adrenal vein localization by 3D real-time volume-rendering CTA before laparoscopic nephrectomy in living renal donors. AB - BACKGROUND: We investigated whether the left adrenal vein could be consistently localized on three-dimensional (3D) real-time volume-rendering computed tomographic angiographic (CTA) mapping in a group of living renal donors before laparoscopic nephrectomy. METHODS: Sixty-six consecutive potential renal donors were referred for CTA vascular mapping before laparoscopic donor nephrectomy. Thirty-three patients were examined on a single-detector helical CT scanner and the other 33 were examined on a multidetector unit. In each patient, arterial phase and venous phase volumetric data sets were acquired after the intravenous injection of 150 cc of non-ionic contrast material. Three radiologists reviewed the data sets at a free-standing workstation after the application of 3D volume rendering software and reached a consensus on whether the left adrenal vein was visualized and, when seen, its position relative to the abdominal aorta. RESULTS: The left adrenal vein was found in 92.5% of the 66 donors (91% in the single detector group and 94% in the multidetector group). The junction of the left adrenal vein and left renal vein averaged 5.2 mm (range = 0-13 mm) from the left lateral wall of the abdominal aorta. CONCLUSION: Three-dimensional real-time volume-rendering CTA with single-detector and multidetector scanners permits consistent localization of the left adrenal vein in more than 90 % of prospective living renal donors. PMID- 11503099 TI - Sickle-beta-thalassemia and splenic calcification. PMID- 11503100 TI - Population-based developmental outcomes monitoring for high-risk infants is imperative. PMID- 11503101 TI - Focal small bowel perforation: an adverse effect of early postnatal dexamethasone therapy in extremely low birth weight infants. AB - OBJECTIVE: We tested the hypothesis that early postnatal dexamethasone (EPD) increases the risk of focal small bowel perforation (FSBP) in extremely low birth weight (ELBW) infants. STUDY DESIGN: The techniques of meta-analysis were applied to studies evaluating EPD, which we identified through a systematic literature search. Studies were included if they were randomized, placebo-controlled trials of EPD, enrolled infants with birth weights < or =1000 g, and reported FSBP as an outcome variable. The Breslow-Day test was used to assess for homogeneity and a summary odds ratio was calculated using the Mantel-Haenszel exact method. RESULTS: Four studies, with a pooled sample of 1383 infants, were included in the primary analysis. The Breslow-Day test showed a p-value of 0.61, indicating homogeneity among the studies. FSBP was significantly higher in EPD treated infants [odds ratio 1.91, 95% confidence interval (CI) 1.21, 3.07; p=0.004]. CONCLUSION: EPD increases the risk of FSBP in ELBW infants. PMID- 11503103 TI - Thermal stability and transition studies with a hybrid warming device for neonates. AB - OBJECTIVE: The use of both warmer beds and incubators is common in neonatal intensive care units (NICU), and transferring between these two warming devices is a routine and necessary event. This study was designed to evaluate the efficacy of a new hybrid-warming device, the Versalet, in transitioning a preterm animal from a warmer bed to an incubator mode and back. STUDY DESIGN: Nine premature lambs were randomized, following delivery, to receive thermal support from a conventional warming bed and an incubator (control group), or from the Versalet (study group) in the warmer bed and incubator modes. Core and various surface temperatures, as well as physiological parameters were measured first during warming in the radiant warmer bed mode, Versalet or Resuscitaire and then during transition to the incubator mode, Versalet or Isolette, and then back to the warmer bed mode. RESULTS: The animals remained stable during all the transitions. Despite careful planning, adverse events occurred in the control group during transfers. There were no significant differences in the temperature or physiologic profiles during any of the transitions in either group. CONCLUSION: Compared with the standard warming technique used in NICUs (separate warmer bed and incubator), the Versalet provides similar thermal and cardiovascular stability without adverse events during transition to different modes of warming. The degree to which this device would contribute to ease of management and improved outcomes in humans needs to be evaluated in a clinical trial. PMID- 11503102 TI - Training pediatric house staff in evidence-based ethics: an exploratory controlled trial. AB - OBJECTIVE: To evaluate an educational intervention in evidence-based ethics (emphasizing clinical knowledge, epidemiologic skills, and recognition of ethical issues) administered to house staff before rotating through our neonatal intensive care unit. STUDY DESIGN: A controlled trial of 64 pediatric house staff assigned to alternating control and intervention rotations. Questionnaires were administered at the end of the rotation. RESULTS: Some benefits of the intervention were observed. However, a large percentage of intervention and control house staff substantially overestimated (>1.25 correct value) predischarge mortality (23% vs. 55% of house staff; p<0.02), mortality or major morbidity (74% vs. 46% of house staff; p=0.04), and cerebral palsy rates (70% vs. 87%; p=0.12). Neither group cited many methodological criteria for evaluating follow-up studies (3.3 vs. 2.4 criteria; p=0.05) or ethical issues considered in treatment recommendations for extremely premature infants (3.1 vs. 2.8 issues; p=0.35). CONCLUSION: Improved house staff training in evidence-based ethics is needed. PMID- 11503104 TI - Significance of meconium-stained amniotic fluid in the preterm population. AB - OBJECTIVE: Numerous studies have assessed the significance of meconium-stained amniotic fluid (MSAF) at term. However, to date, there has been very little documentation on the incidence and significance of meconium in the preterm population. Our objective was to define the incidence of MSAF in patients delivering prematurely (<37 weeks) and examine its association with underlying fetal acidosis, Apgars and admission to the neonatal intensive care unit (NICU). METHOD: All patients delivering at a single tertiary care center between June 1994 and September 1997 were reviewed for the presence of meconium and gestational age <37 weeks at delivery. Maternal demographics and birth outcomes including cord gases, Apgar scores and admission to the NICU were collected. Exclusion criteria included multiple gestations, breech presentations, fetal anomalies and patients not in labor. RESULTS: Out of a total of 9570 patients there were 506 (5.3%) preterm births meeting the inclusion criteria, of whom 24 (4.8%) had MSAF noted either during labor or at delivery. Comparing the preterm group with and without meconium, there were no differences in maternal age, gravidity, rate of Cesarean section, or gestational age at delivery. Cord pH (7.27 meconium vs. 7.29 no meconium) and base excess (-5.1 meconium vs. -4.0 no meconium) were similar in both groups. There were no clinically significant differences in mean Apgar scores at 1 and 5 minutes. However, an increased number of NICU admissions were noted in the group with meconium (75% vs. 53%, p=0.04). CONCLUSION: The incidence of meconium staining of the amniotic fluid in labor in the preterm population is less than 5% and by itself is not a significant marker of fetal acidosis. PMID- 11503105 TI - Labor epidurals improve outcomes for babies of mothers at high risk for unscheduled cesarean section. AB - CONTEXT: Epidural placement for labor in the general population of laboring women is associated with increased incidence of operative deliveries, prolongation of labor, and may be associated with an increased cesarean section rate. The risks and benefits associated with epidural placement for labor in the subpopulation of mothers at high risk for cesarean section have not been studied. OBJECTIVE: To determine if a population of mothers and babies at high risk for cesarean section will have improved outcomes with labor epidural placement. DESIGN: A decision and cost analysis examining epidural placement for labor on a population of women who are at high risk for unscheduled cesarean section and may benefit from scheduled cesarean section as determined by threshold analysis was performed. Outcomes and probabilities were determined through analysis of the Department of Defense's 1996 National Quality Management Program (NQMP) Birth Product Line data set containing more than 7000 deliveries. Outcomes were defined using variables comprised of all documented conditions that occurred during the peripartum and neonatal hospitalizations. The 1997 NQMP data set was used to validate the results. SETTING: Military Treatment Facilities throughout the United States and abroad and civilian facilities in the United States providing care to military dependents. PATIENT POPULATION: Active duty and dependent pregnant women and babies. RESULTS: About 8% of mothers in this patient population were found to be at high risk for cesarean section. The decision and cost analyses showed that babies of the high risk mothers who received epidurals for labor had better clinical outcomes (p<0.05) and the procedure was cost neutral (p=0.23). The procedure did not increase the frequency of cesarean section, and there was no effect on maternal outcomes scores. These results were confirmed by the validation study. CONCLUSIONS: There is a sizable subpopulation of women at high risk for cesarean section whose babies may have better outcomes with epidural placement with no sacrifice in maternal outcomes or costs. PMID- 11503106 TI - Nosocomial coagulase negative staphylococcal (CoNS) catheter-related sepsis in preterm infants: definition, diagnosis, prophylaxis, and prevention. AB - Nosocomial infections with coagulase negative staphylococcus (CoNS) are a frequent and significant cause of morbidity in the preterm infant. Infections diagnosed after the first 72 hours of life are arbitrarily deemed to be "nosocomial." There are many difficulties encountered in efforts to evaluate and compare nosocomial sepsis in the NICU. An issue of primary concern is the lack of uniformity in the definition of sepsis in the NICU. Based on the frequency of positive blood cultures in infants less than 1000 g, it appears reasonable to evaluate methods for the prevention of nosocomial sepsis. These include prophylactic antibiotic administration, antiseptic impregnated catheters, and the use of an antibiotic lock technique. PMID- 11503107 TI - Decisions regarding resuscitation of the extremely premature infant and models of best interest. AB - Differences concerning the care of an extremely premature infant may stem from alternative points of view on how to determine the infant's best interest. These alternatives are illustrated by differences between recently published statements by the American Academy of Pediatrics (AAP) Committee on Fetus and Newborn (COFN) and the Committee on Bioethics (COB). The statements agree that a goal of neonatal medicine is to minimize both under- and overtreatment of the extremely premature infant, and advocate that the decision-making process ought to be based on the concept of the premature infant's best interest. However, the two AAP Committees appear to diverge in how they operationalize the concept of an infant's best interest. The COFN adopts a process consistent with an "expertise" model of best interest, while the COB process is consistent with a "negotiated" model. In the "expertise" model, medical re-evaluation of the infant's status, the best outcome data available, and the treating physician's best medical judgment determine best interest. This model limits parental and societal input, and can lead a physician to act paternalistically. In the "negotiated" model, best interest is determined by outcome data and physician assessment, as well as the moral value of an outcome. This model maximizes parental input, accepts physicians as moral agents, and respects social influence in a decision. It is important to clarify one's model of best interest to help understand the differences of opinion regarding decisions based on best interest. The negotiated model of best interest is a more ethically appropriate model to approach decision making. PMID- 11503109 TI - A neonate with Loffler syndrome. AB - We report a neonate-boy with pulmonary infiltrates and peripheral blood eosinophilia. He was noted to have abnormal pulmonary infiltrates on a chest X ray film taken on day 8 after birth when he had vomiting. He had not such symptoms as cough or dyspnea. In routine laboratory studies, eosinophilia was noted. Radiographic changes were transient and disappeared by day 25. Eosinophilia was also transient and gradually returned to normal level by 2 months. Loffler syndrome is very rare in neonates and its diagnosis is often made fortuitously likely in this case. PMID- 11503110 TI - Congenital cytomegalovirus infection (CMV). PMID- 11503112 TI - The effect of pulsed irrigation on the incidence of heterotopic ossification after total hip arthroplasty. AB - Heterotopic ossification (HO) is a common complication of total hip arthroplasty (THA). Pulsed lavage is being used with increasing frequency for THA. A prospective randomized, double-blind trial was initiated to determine if pulsed lavage affected the incidence of HO. A total of 94 THAs in 91 patients were analyzed. No significant difference in the incidence of HO was found between the 2 groups. Hypertrophic osteoarthritis was found to be a significant risk factor for HO. The findings suggest that the osteogenic precursor cells thought to be involved in the pathogenesis of HO possibly are derived from within the local soft tissues in the proximity of the hip joint. PMID- 11503113 TI - Femoral remodeling after porous-coated total hip arthroplasty with and without hydroxyapatite-tricalcium phosphate coating: a prospective randomized trial. AB - We prospectively assessed femoral bone remodeling using dual-energy x-ray absorptiometry for 2 years after total hip arthroplasty. Thirty-nine hips were randomized to receive a titanium proximally porous-coated femoral component with or without hydroxyapatite-tricalcium phosphate coating. Although both stems resulted in alterations in the periprosthetic bone mineral density, the hydroxyapatite-tricalcium phosphate coated stems had significantly less femoral bone loss than the uncoated stems at 2-year follow-up. This reduced femoral bone loss may provide short-term and long-term advantages over noncoated stems. PMID- 11503114 TI - Cementless porous-coated total knee arthroplasty: 10-year results in a consecutive series. AB - We report the results of 114 AGC 2000 porous-coated, cementless total knee arthroplasties (TKA) performed consecutively in 102 patients during the period 1984-1986. After 10 years, 58 TKAs in 52 patients were evaluated with patient assessment, Hospital for Special Surgery knee score, weight-bearing radiographs done under fluoroscopic control, and survivorship analysis. All dropouts within the first 9 years were patients dying with a functioning TKA except 1 revision secondary to a supracondylar fracture after 8.5 years. Of the patients, 53 (92%) were satisfied or very satisfied with their TKA, and 55 (95%) of the knees were rated good or excellent. There was no pain in 53 knees, and the median knee flexion was 110 degrees. Six radiolucencies >1 mm were found beneath parts of the tibial component, and 5 radiolucencies were seen beneath the femoral component. None had progressed compared with the 5-year follow-up, and in all cases trabeculae could be seen reaching the prosthetic component. No migrations had occurred since the 5-year follow-up. No obvious joint space reduction was seen. Osteolysis presenting as an isolated cyst was found in 1 knee in the lateral tibial condyle and was not progressive. Two tibial components had been revised because of aseptic loosening and 1 because of septic loosening, all within the first 3 years. No femoral or patellar components were revised. The cumulative prosthesis survival rate after 10 to 11 years was 97%. When pain and radiographic loosening also were considered, the success rate was 87%. Cementless insertion of a nonmodular, porous-coated TKA resulted in a long-term durable bone-prosthesis interface. The flat-on-flat articulation did not result in catastrophic polyethylene wear or osteolysis within the first 10 years. PMID- 11503115 TI - Reconstruction of segmental defects during revision procedures of the acetabulum with the Burch-Schneider anti-protrusio cage. AB - Sixty-three Burch-Schneider rings in 62 patients with segmental or combined defects were followed up after a mean of 5.45 years. The mean Harris hip score was 74.9 points. Aseptic loosening occurred in 3 cases, and septic loosening occurred in 2 cases. All cases of aseptic loosening had Paprosky stage IIIb defects and defects of the posterior column. Radiologic migration analysis showed increased migration with increase in the Paprosky defect stage, defects of the posterior column, and cranial defects. The cage has the lowest migration and loosening rate in cases with medial defects or anterior column defects. Surgical treatment of defects of the posterior column is associated with an increased rate of aseptic loosening, whereas implantation with cranial defects shows a higher migration rate with no significant increase in loosening. PMID- 11503116 TI - The Harris hip score: comparison of patient self-report with surgeon assessment. AB - Outcome evaluations are of primary concern in contemporary medical practice. Questionnaires are being used increasingly to provide input data for such outcomes evaluation. This study comprised 50 primary total hip arthroplasties in 36 patients who had undergone the procedure at least 12 months before enrollment. Each patient completed a self-report Harris Hip Score (HHS) 30 days before a formal evaluation by an independent orthopaedic surgeon that included a HHS. Comparison was made between the completed responses to the individual items on the self-report HHS and surgeon-assessed HHS. Concordance of item response and kappa statistic were calculated. Overall the self-report and surgeon-assessed HHS showed excellent concordance. The results of this study support the use of the HHS as a self-report instrument. PMID- 11503117 TI - Does implant selection affect outcome of revision knee arthroplasty? AB - We reviewed 139 consecutive femoral or tibial revision knee arthroplasties to determine if the outcome of revision knee arthroplasty using revision implant systems was superior to revisions using primary implant systems. Group 1 (n = 42) consisted of revisions performed with implants designed for primary total knee arthroplasty. Group 2 (n = 42) consisted of revisions performed with modified primary components. Group 3 (n = 55) consisted of revisions performed with components specifically designed for revision arthroplasty. The implant status was known in 123 of 139 knees at a mean follow-up of 7 years (range, 5-12 years). The implant-related failure rate, defined as reoperation requiring component revision or removal, was 26% for group 1, 14% for group 2, and 6% for group 3 (P<.05). Revision implants exhibited superior performance and durability despite their use in more difficult reconstructions. The improved longevity of revision implants justifies the evolution of modular revision components. PMID- 11503118 TI - The Harris-Galante total hip arthroplasty: a minimum 8-year follow-up study. AB - A total of 93 Harris-Galante Porous I (HGP- I) prostheses were analyzed with a mean follow-up of 10.2 years. Thigh pain (28 hips) correlated with unstable fixation (P<.005), female gender (P<.045), young age (P<.05), poor distal femoral fill (P<.002), subsidence (P<.0001), and osteolysis (P<.05). Thirteen stems and 6 metallic shell cups were revised. Kaplan-Meier analysis showed 13-year survival rates of 92.5% +/- 5.8% for the metallic shell cup, 79.7% +/- 13% for the HGP-I cup, and 76.3% +/- 14.0% for the stem. Five polyethylene liners were changed because of wear >1 mm. Radiographic loosening occurred in 8 cups. Radiographic bone ingrowth was present in 54 stems, stable fibrous fixation was present in 24 stems, and unstable fixation was present in 15 stems. The mean polyethylene wear was 0.17 mm/y. Eleven hips (11.8%) had acetabular osteolysis, and 24 (25.8%) had femoral osteolysis, the latter being more frequent in unstable stems (P<.007). The HGP-I metallic cup shows better clinical and radiographic results than the stem, which frequently is associated with pain, unstable fixation, and osteolysis. Excessive polyethylene wear is frequent. PMID- 11503119 TI - Should total hip arthroplasty femoral components be designed to subside? A radiostereometric analysis study of the Charnley Elite and Exeter stems. AB - The Charnley Elite and the Exeter stems have different design concepts: The former is designed not to subside, whereas the latter is expected to subside. This radiostereometric analysis study compares the early migration of the 2 stems. For both implants, the 1st year migration was about 4 times faster than the 2nd year. The Exeter migration was predominantly distal (1 mm/y in the 1st year). It also showed slight collapse into valgus, and the head migrated slowly posteriorly (0.3 mm/y in the 1st year). In contrast, the Elite had slow distal migration (0.2 mm/y in the 1st year) and rapid posterior head migration (0.8 mm/y in the 1st year). Four Elites and no Exeters had rapid posterior head migration rates (mean 2.8 mm/y in the 1st year and 0.8 mm/y in the 2nd year). The Elite and the Exeter stems have fundamentally different early patterns of migration, which affect their long-term function; 20% of the Elites and none of the Exeters had rapid posterior head migration in the 1st year and the 2nd year and are likely to fail early. Polished, collarless, tapered designs, such as the Exeter, may be more forgiving than conventional stems designed not to subside. PMID- 11503120 TI - Total knee arthroplasty without patellar resurfacing in isolated patellofemoral osteoarthritis. AB - Thirty-one patients (33 knees) with symptomatic patellofemoral osteoarthritis and minimal tibiofemoral changes underwent LCS total knee arthroplasty without patellar resurfacing. Average age was 73 years (range, 58-89 years) with a female to-male ratio of 5:1. Average follow-up was 20 months (range, 12-40 months). All except 4 patients had grade 3 or 4 patellofemoral osteoarthritis. Preoperatively, all patients had significant knee pain. Sleep disturbance was reported in 21 patients. All but 10 patients required walking aids. Average range of motion was 108 degrees (range, 80-125 degrees ). At latest review, 21 knees were pain-free, the remaining 12 knees being described as having only occasional knee pain. Two patients continued to have night pain. Average range of motion was 104 degrees (range, 70-135 degrees ). Lateral patellar tilt improved in all but 5 knees by an average of 7 degrees (range, 1-26 degrees ). Patellar congruency improved in all but 3 knees by an average of 18% (range, 3-63%). None of the patients to date have required revision surgery. Total knee arthroplasty without patellar resurfacing is an effective option in older patients with isolated patellofemoral osteoarthritis. PMID- 11503121 TI - Comparative study of the activity of total hip arthroplasty patients and normal subjects. AB - The walking activity of normal subjects and total hip arthroplasty (THA) patients from the Wrightington Hospital for Joint Disease and The General Infirmary at Leeds was assessed by means of electronic pedometers. The principal objectives were to establish the extent to which joint arthroplasty patients recover their activity relative to normal subjects and to establish the number of loading cycles to which prostheses should be subjected in joint simulator studies of implant performance. A further objective was to establish an experimental procedure for the assessment of the role of activity in contributing to the well known scatter in the measurements of femoral head penetration into acetabular cups in in vivo studies of implant performance. The last-mentioned issue is addressed in another article ([1]). The walking activity of 2 normal subjects of disparate ages was assessed during 1 full year. It was concluded that fair estimates of activity could be achieved by recording pedometer readings during successive 2- to 4-week periods. This approach was adopted in the full assessment of the walking activity of cohorts of normal subjects and THA patients. Linear regression expressions relating the number of steps taken daily and the annual number of loading cycles on each leg to age are presented for normal subjects and THA patients. In all cases, activity declines with age, but it is shown that total joint arthroplasty is not at all restrictive on walking activity-a remarkable testimony to the efficacy of total joint arthroplasty. Attention is drawn, however, to different levels of activity of THA patients recorded in the present United Kingdom study and a similar survey conducted in California. PMID- 11503122 TI - The effect of activity levels of total hip arthroplasty patients on socket penetration. AB - Survivorship of total hip arthroplasties (THAs) has been linked to penetration of the femoral head into the polyethylene acetabular cup and to polyethylene wear. The activity level of patients with THAs is considered to be an important factor affecting wear, and the purpose of this study was to explore the relationship between activity as recorded by pedometers and cup penetration. The measurement of daily activity levels of normal subjects and THA patients of various ages are discussed in another article ([1]). Subjects were monitored continuously during 2 to 4-week periods using simple pedometer devices. Patients (n = 54; mean age, 58 years) from the Centre for Hip Surgery at Wrightington Hospital for whom the penetration data also were available were included in the study. The average activity level for the patient group was 1.426 million loading cycles on each hip joint per year. Radiographic penetration measurements were compared for 81 hips in 54 patients with a mean follow-up of 13.1 years (range, 1.3-26.4 years). The overall correlation of penetration with implantation period is known to be poor, however, and did not improve significantly when the penetration was plotted against a new parameter that took account of not only implant life, but also the level of activity and patient weight. The considerable scatter of penetration levels was noted to increase with increasing implantation period, which indicates that in the multifactorial problem of polyethylene wear, other factors, particularly femoral head surface finish or polyethylene deterioration, may predominate. PMID- 11503123 TI - The influence of technique on fixation of primary total hip arthroplasty in patients with rheumatoid arthritis. AB - This is a prospective, consecutive study of 98 total hip arthroplasties implanted by 1 surgeon in 66 patients with rheumatoid arthritis. The mean follow-up time was 7.4 years (range, 2-13 years). All 98 acetabular components were uncemented titanium fiber metal-coated components fixed with multiple screws. Sixty-five hips had bulk or cancellous allograft for protrusio acetabuli. Following a prospective protocol based on patient age, 51 hips had an uncemented femoral component, and 47 hips had a cemented femoral component. Using the Harris Hip Score, 30 hips were rated as excellent; 44, good; 15, fair; and 9, poor. There were no early deep infections. One hybrid hip was removed for late metastatic infection at 7 years. Radiographic evaluation of 98 acetabular components showed 1 case of septic loosening, 2 cases of aseptic loosening (1 patient asymptomatic), and 1 case with severe wear and ischial osteolysis. None of the 47 cemented femoral components subsided, and osteolysis was seen in only 3 femora (7%). Of the 51 uncemented femoral components, subsidence occurred in 7 hips (14%), and osteolysis occurred in 15 hips (30%). Uncemented acetabular components have a high rate of success in patients with rheumatoid arthritis who have a total hip arthroplasty. There is a high rate of subsidence and osteolysis, however, with first-generation cementless femoral components. PMID- 11503124 TI - The effect of surgeon experience on component positioning in 673 Press Fit Condylar posterior cruciate-sacrificing total knee arthroplasties. AB - Component angles of 673 Press Fit Condylar (PFC) total knee arthroplasties were measured from standard short-leg radiographs. The femoral and tibial resections were performed with intramedullary and extramedullary instrumentation. The mean coronal tibial component angle was 88.59 degrees (SD, 2.28 degrees; range, 78-98 degrees ), with 17.1% having values <87 degrees and 1.9% having values >93 degrees. The mean coronal femoral component angle was 97.43 degrees (SD, 3.44 degrees; range, 84-115 degrees ), with 9.1% having values <94 degrees and 13.1% having values >100 degrees. An ideal tibiofemoral angle of 4 degrees to 10 degrees of valgus was achieved in 75.3% of patients, being <4 degrees in 18.6% and >10 degrees in 6.1%. Alignment was not significantly different between consultant and trainee surgeons. Although varus positioning of the tibial component was the commonest error, the wide range of femoral component angles signifies problems with standard intramedullary femoral guides. PMID- 11503125 TI - Geometric analysis of potential error in using femoral intramedullary guides in total knee arthroplasty. AB - Proper component orientation is necessary for the long-term success of a total knee arthroplasty. Femoral component placement has used jigging systems that are based on intramedullary and extramedullary guides. The intramedullary system has been shown to foster more accurate and reproducible placement of the femoral component. This study analyzed and quantified the potential error of the intramedullary guide system. Geometric data of the human femur were obtained from radiographs of 45 cadaver femora in the anteroposterior and lateral views. The correct entry point of the intramedullary rod in the distal femur is the midaxial line of the femoral canal on anteroposterior and lateral radiographs. The position of this point in relation to the mediolateral and anteroposterior dimensions of the distal femur was calculated as a ratio. The average calculated ratios were anteroposterior, 0.53 offset medially; lateral, 0.32 offset anterior. Mathematical models were constructed to quantify the potential varus valgus and flexion extension error of the guide rods. Variables investigated were entry point location, guide rod diameter, guide rod length, and rotation. The surgeon must be cognizant of these potential sources of error to maximize the accuracy of the femoral intramedullary jigging system. PMID- 11503126 TI - Dependency of cement mantle thickness on femoral stem design and centralizer. AB - Deficient cement mantles may be detrimental with regard to long-term outcome of cemented femoral stems. We performed a cadaver study on 48 left femora with 4 different stem designs (1 anatomic, 3 straight) to study the influence of stem design, centralizer, and femur type on cement mantle thickness. A radiographic and microradiograhic analysis was done. Overall, 88% of stems were aligned within 1 degrees of neutral in the frontal plane. In Gruen zones 1 through 7, we measured 24 thin cement mantles (<2 mm) in 19 specimens with no correlation to stem design or zone. In the sagittal plane, typical areas of thin cement mantles were identified in Gruen zones 8 and 9 (n = 39) and 12 (n = 21). The anatomic stem design carried the lowest risk (54%) of producing a thin cement mantle proximally in Gruen zones 8 and 9. The risk for straight stem designs was >90%. Straight stems without centralizer showed the highest risk of thin cement mantles in Gruen zone 12 (93%). Centralizers were efficient to prevent thin cement mantles in zone 12 but had no effect proximally. Lateral radiographs are essential to allow for adequate radiographic assessment of the cement mantle and stem alignment. There is a high risk of producing thin cement mantles in Gruen zones 8 and 9, in particular when straight stems are used. Posterior canal entry and low neck osteotomies are essential. Anatomic stems respect the anatomy, allow for more even cement mantles, minimize the risk of thin cement mantles without the use of centralizers, and may be considered in the femur with marked proximal bow. PMID- 11503127 TI - A new pin-on-disk wear testing method for simulating wear of polyethylene on cobalt-chrome alloy in total hip arthroplasty. AB - Hip simulator studies show that the wear of ultra-high molecular weight polyethylene against a cobalt alloy head depends on the wear path, especially the combination of a predominantly linear wear direction on which is superimposed motions in different directions. We postulated that multidirectional motion was necessary to generate realistic wear rates in pin-on-disk testing. To assess this hypothesis, a new pin-on-disk tester was developed, capable of unidirectional and bidirectional motion. Unidirectional motion produced no detectable wear. The rectangular motion produced wear rates, surface morphologies, and wear particles consistent with human acetabular specimens. The results for 1 Hz and 2 Hz were similar. PMID- 11503128 TI - Acetabular revision with impacted freeze-dried cancellous bone chips and a cemented cup: a report of 7 cases at 5 to 9 years' follow-up. AB - The long-term results of bone impaction grafting with fresh-frozen femoral head allografts and a cemented cup are favorable. Because of intermittent shortage of fresh-frozen femoral heads at our local bone bank, we used processed freeze-dried bone in 7 acetabular revisions operated between 1989 and 1994. All 7 consecutive patients were followed annually. At final review (March 2000), 1 patient had died after 8.5 years of follow-up of a cause not related to the surgery. In 1 hip, a rerevision was performed for septic loosening 5 years after the previous septic loosening. Radiographically the freeze-dried allografts seemed to incorporate in all cases but the reinfected one; progressive radiolucent lines were not seen, although 1 case had a stable line in 1 zone. The overall survival rate for the 7 acetabular reconstructions at an average follow-up of 7 years (range, 5-9 years) was 86%. At midterm follow-up, there was no aseptic loosening. In this limited case report, the results at midterm for freeze-dried allograft bone chips in acetabular reconstructions are acceptable. PMID- 11503129 TI - Superior mesenteric vein tear with total hip arthroplasty. AB - Total hip arthroplasty imparts significant physical forces on the patient at the time of surgery. We report a case of an injury to the superior mesenteric vein that is thought to have occurred at the time of impaction of the acetabular component of a total hip arthroplasty. This complication has been reported previously only from high-energy nonpenetrating trauma, such as motor vehicle accidents. PMID- 11503130 TI - Wear debris from total hip arthroplasty presenting as an intrapelvic mass. AB - A granulomatous reaction to the small particles worn off from an implanted prosthesis is a common complication in arthroplasty. The chronic inflammatory response to the wear debris from either the polyethylene or the metal components is thought to be responsible for osteolysis, implant loosening, and finally mechanical failure. Formation of an extra-articular granulomatous tissue mass is uncommon, however. We report a case of a huge granulomatous mass that presented as an intrapelvic tumor. The prosthesis was an Anatomical Medullary Locking (AML) hip arthroplasty with cementless fixation. The polyethylene insert was manufactured by ram extrusion and sterilized with gamma irradiation in air. Fragments of the insert were among the contents in the cystic mass together with black metallic debris. There was no loosening of the prosthesis, and the acetabulum floor was preserved. PMID- 11503131 TI - Total hip arthroplasty in young adults after failed triple innominate osteotomy. PMID- 11503133 TI - Is antiphospholipid antibody determination clinically relevant to the vasculitides? PMID- 11503134 TI - Lack of association between antiphospholipid antibodies and thrombocytopenia in patients with Wegener's granulomatosis. AB - BACKGROUND AND OBJECTIVE: In patients with Wegener's granulomatosis (WG), thrombocytopenia is less common than thrombocytosis. An increased prevalence of antiphospholipid antibodies (aPL), which is associated with thrombocytopenia, has been noted in patients with WG. The aim of this study was to examine the relationship between thrombocytopenia and aPL in patients with WG. METHODS: Thrombocytopenic episodes were searched for in a random sample of 83 patients with WG. Stored sera obtained during thrombocytopenia, which was defined as platelet count below 130 x 10(9)/L, were examined by 2 different enzyme-linked immunosorbent assays (ELISA) for IgG and IgM anticardiolipin antibodies (aCL) and for IgG antiphosphatidylserine antibodies (aPS). Screening for lupus anticoagulant was performed by use of activated partial thromboplastin time (aPTT). Results were compared with the prevalence of aPL in 20 consecutive nonthrombocytopenic patients with WG. RESULTS: Six cases with thrombocytopenic episodes were found in the group of 83 patients with WG. Increased IgG and IgM aCL were detected in 1 patient, who also had elevated IgG aPS. A positive test result solely for IgM aCL was found in another patient. These findings were consistent in both ELISA for aPL. Five patients were being treated with cyclophosphamide when thrombocytopenia occurred. In the group of nonthrombocytopenic patients with WG, elevated IgG aCL and IgG aPS were consistently detected in 1 patient in both ELISA. Three other patients had positive results in single tests, which were not confirmed by the second assay. In all patients, aPTT was normal. CONCLUSIONS: Thrombocytopenia is a rare finding in patients with WG. A similar prevalence of aPL in thrombocytopenic and nonthrombocytopenic patients with WG provides no evidence that aPL play a major role in the pathogenesis of these events. Thrombocytopenia in WG is more likely caused by the myelotoxic effect of preceding cyclophosphamide treatment. We found a frequency of aPL in WG that exceeds frequencies seen in the general population but does not approximate those detected in systemic lupus erythematosus and closely related disorders. Semin Arthritis Rheum 31:4-11. PMID- 11503135 TI - Antiphospholipid antibodies and thrombophilic factors in giant cell arteritis. AB - OBJECTIVES: To evaluate the prevalence of thrombophilic risk factors known to induce intravascular clotting and to assess their relationship with ischemic manifestations in giant cell arteritis (GCA). METHODS: Eighty consecutive patients with established GCA were included: 36 with isolated temporal arteritis (TA), 14 with isolated polymyalgia rheumatica (PMR), and 30 with TA and PMR. Forty-four patients (67%) had ischemic phenomena due to GCA. Twelve patients (15%) had thrombotic events unrelated to GCA (6 strokes, 5 deep venous thrombosis, and 1 myocardial infarction). A control group of 100 age- and sex matched individuals without autoimmune disease, bleeding disorders, thrombosis, or clinical picture of TA or PMR also was analyzed. All participants were tested for the antiphospholipid antibody (aPL) profile, protein C, protein S, antithrombin activity, factor V Leiden mutation, and prothrombin gene G20210A mutation. We also studied fibrinolysis parameters: plasminogen, tissue-type plasminogen activator (t-PA) antigen, t-PA activity, type-1 plasminogen activator inhibitor (PAI-1) antigen, PAI-1 activity, and the 4G/5G polymorphism of the promoter region of the PAI-1 gene. RESULTS: Eleven patients (18%) tested positive for lupus anticoagulant, 24 (30%) for anticardiolipin antibodies, 9 (11%) for anti-beta 2-glycoprotein I antibodies, and 29 (36%) for antiprothrombin antibodies. No relationship was found between these autoantibodies and ischemic manifestations. None of the patients had decreased protein C, protein S or antithrombin activity. Two patients and 2 controls were heterozygous for factor V Leiden, and only 1 patient and 2 controls were heterozygous for the prothrombin gene G20210A mutation. No statistically significant correlation was found between any thrombophilic factor and GCA-related or GCA-unrelated ischemic events. CONCLUSION: GCA patients have a high prevalence of aPL that is not related to ischemic manifestations. Moreover, GCA-related or GCA-unrelated ischemic manifestations do not appear to be due to congenital thrombophilic risk factors. Semin Arthritis Rheum 31:12-20. PMID- 11503136 TI - Bromocriptine in rheumatic and autoimmune diseases. AB - BACKGROUND AND OBJECTIVES: Multiple lines of evidence support the concept that the anterior pituitary hormone prolactin has a pathogenic role in rheumatic and autoimmune diseases including, but not limited to, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), Reiter's syndrome, psoriatic arthritis, and uveitis. Conversely, the dopaminergic agonist bromocriptine appears to have therapeutic effects through suppression of pituitary prolactin secretion and, perhaps, through actions on peripheral dopamine receptors. This article reviews the experimental and clinical data supporting the therapeutic use of bromocriptine as a nonstandard or adjunctive therapy in rheumatic and autoimmune diseases. METHODS: Data addressing the potential therapeutic role of bromocriptine in rheumatic and autoimmune diseases, as well as frequently associated comorbidities, was accumulated from the author's work, online literature search of the National Library of Medicine, and references from these identified publications. RESULTS: There have been a number of clinical therapeutic trials using 2.5 to 30 mg of bromocriptine per day in a single or divided dose, which have shown efficacy with minimal side effects in the treatment of rheumatic and autoimmune diseases. In RA, bromocriptine administration has induced immunosuppression of several immune parameters and has been associated with improvements in morning stiffness, grip strength, numbers of swollen/painful joints, and the Health Assessment Questionnaire disability index. In two blinded studies, bromocriptine reduced the number of SLE flares and was as effective as hydroxychloroquine in reducing lupus disease activity indices, respectively. In case reports, bromocriptine has been used successfully in the treatment of Reiter's syndrome enthesopathy and psoriatic arthritis. The potential efficacy of bromocriptine in the treatment of uveitis and multiple sclerosis is suggested but remains to be verified. CONCLUSIONS: Double-blind, placebo-controlled studies are limited, but clinical observations and trials support the use of bromocriptine as a nonstandard primary or adjunctive therapy in the treatment of recalcitrant RA, SLE, Reiter's syndrome, and psoriatic arthritis and associated conditions unresponsive to traditional approaches. Additional investigation is needed to verify this conclusion and extend preliminary results. RELEVANCE: In patients with rheumatic and autoimmune diseases, bromocriptine may be a relatively safe and efficacious alternative therapy. Semin Arthritis Rheum 31:21-32. PMID- 11503137 TI - Bone morphogenetic proteins in the development and healing of synovial joints. AB - OBJECTIVES: To review current knowledge of the role of bone morphogenetic proteins (BMPs) in joint formation and how this may be relevant to healing in adult joints. METHOD: Review of published literature using a search of the PubMed database (1966 to 2000) made available by the National Library of Medicine. Additional articles of historical interest were identified from the bibliographies of published literature. RESULTS: BMPs and a related family, the growth and differentiation factors (GDFs), are stimulators of bone and cartilage formation in the developing skeleton. They, together with their antagonists, play key roles in the specification of the joint site and cavitation of synovial joints during embryonic development. Disruption of the GDF-5 gene in mice and humans is associated with abnormal joint formation. In situ hybridization studies have shown that BMPs are expressed during formation of synovial joints in the embryo. However, excessive BMP activity leads to obliteration of joints because of cartilage overgrowth. BMPs are being considered as therapeutic agents to stimulate healing of articular cartilage after damage. Evidence suggests that BMPs are present in adult joints and have roles in healing and maintenance. However, inflammatory cytokines and growth factors present in damaged joints modulate the actions of BMPs. CONCLUSIONS: BMPs, and in particular GDF-5, are involved in synovial joint formation. They may also have effects on the maintenance and healing of adult joints, but factors present after damage may alter their effectiveness. RELEVANCE: Articular cartilage heals poorly after damage. BMPs may be useful therapeutically to stimulate healing of damaged articular cartilage. Increased knowledge of their role in joint formation will improve understanding of how to use them. Semin Arthritis Rheum 31:33-42. PMID- 11503138 TI - Acute bacterial arthritis caused by group C streptococci. AB - OBJECTIVES: To summarize the characteristics of and clinical experience with joint infection by group C streptococcus. METHODS: Two new cases of acute bacterial arthritis due to group C streptococci are reported, and a MEDLINE search was performed, which located 22 additional cases. RESULTS: Mean age (+/- standard deviation) of all 24 patients was 46 +/- 25 years, and most patients were men (75%). The species most frequently identified was Streptococcus equisimilis (12 cases). Eight patients (33%) had a general risk factor for infection, and the same percentage had some type of arthropathy. Only 3 patients had previous contact with animals. The infection was polyarticular in a third of cases, and the joint most frequently involved was the knee. The majority of patients showed a good response to the treatment with intravenous penicillin. However, 3 patients had functional sequelae, 2 more had residual radiological lesions, and 3 died. CONCLUSIONS: Acute bacterial arthritis due to group C streptococci is a serious but uncommon entity that can affect patients without risk factors. Rapid diagnosis and treatment may improve the outcome. Semin Arthritis Rheum 31:43-51. PMID- 11503139 TI - Elevated blood viscosity in systemic lupus erythematosus. AB - OBJECTIVES: It has been proposed that elevated blood viscosity contributes to atherothrombotic and thromboembolic processes. We evaluated whether there is increased blood viscosity in systemic lupus erythematosus (SLE) that might contribute to cardiovascular complications and reduced tissue perfusion. METHODS: Blood viscosity profiles were evaluated in SLE patients to determine whether rheologic disturbances contribute to the cardiovascular risk profile. Blood viscosity profiles were evaluated in 27 patients with SLE and 46 age- and gender matched controls. Blood viscosity was measured at 37 degrees C and shear rates of 1 s(-1) and 100 s(-1), then corrected to the average hematocrit of the SLE patients. RESULTS: Corrected blood viscosity values were higher in SLE patients than in controls at 100 s(-1) (P =.002). Positive correlations were found between the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index for SLE, which quantifies damage to 12 organ systems and fibrinogen (rho =.39; P =.042) and plasma viscosity (rho =.38; P =.049). CONCLUSIONS: Our data indicate that blood viscosity values at a standard hematocrit are elevated in SLE patients. Further investigations are needed to evaluate whether the increased blood viscosity values in SLE patients contribute to cardiovascular complications and tissue ischemia. CLINICAL RELEVANCE: Because blood viscosity values correlate with the clinical severity of SLE, blood viscosity may contribute to the cardiovascular complications and reduced tissue perfusion in SLE patients. Semin Arthritis Rheum 31:52-57. PMID- 11503140 TI - Biochemical basis of the pharmacologic action of chondroitin sulfates on the osteoarticular system. AB - BACKGROUND: Chondroitin sulfates (CS) are involved in articular metabolism and could be used as therapeutic agents in degenerative articular diseases. OBJECTIVES: To review the published reports describing both the metabolism of glycosaminoglycans (GAG) and their involvement in osteoarticular pathophysiology. METHODS: MEDLINE search for relevant articles and review of cited references. RESULTS: 1) CS are formed of disaccharide units; sulfated galactosamine residues in position 4 or 6 are found in various ratios, depending on the age and the type of tissue. Binding to the core protein through N- and O-linkages leads to aggregates of monomers with high molecular weights. The proteoglycan aggregate exhibits viscoelastic and hydration properties and an ability to interact with the surrounding tissue through electric charges leading to protection of the cartilaginous tissues. 2) CS are synthesized both in chondrocytes and in bone cells by the action of specific glycosyl-transferases; their catabolism occurs in the matrix and involves numerous matrix (metalloproteinases) and lysosomal enzymes. 3) CS are inhibitors of extracellular proteases involved in the metabolism of connective tissues. In addition to their anti-inflammatory effects, CS in vitro stimulate proteoglycan production by chondrocytes; they also inhibit cartilage cytokine production and induce apoptosis of articular chondrocytes. CS increase the intrinsic viscosity of the synovial liquid. 4) In vivo in experimental arthritis, the number and severity of articular symptoms decreases after CS administration. In bones, CS accelerate the mineralization process and bone repair. CONCLUSIONS: All these data suggest that CS play a role in articular and bone metabolism by controlling cartilaginous matrix integrity and bone mineralization. PMID- 11503141 TI - Distribution of androgen and estrogen receptor mRNA in the brain and reproductive tissues of the leopard gecko, Eublepharis macularius. AB - Incubation temperature during embryonic development determines gonadal sex in the leopard gecko, Eublepharis macularius. In addition, both incubation temperature and gonadal sex influence behavioral responses to androgen and estrogen treatments in adulthood. Although these findings suggest that temperature and sex steroids act upon a common neural substrate to influence behavior, it is unclear where temperature and hormone effects are integrated. To begin to address this question, we identified areas of the leopard gecko brain that express androgen receptor (AR) and estrogen receptor (ER) mRNA. We gonadectomized adult female and male geckos from an incubation temperature that produces a female-biased sex ratio and another temperature that produces a male-biased sex ratio. Females and males from both temperatures were then treated with equivalent levels of various sex steroids. Region-specific patterns of AR mRNA expression and ER mRNA expression were observed upon hybridization of radiolabeled (35S) cRNA probes to thin sections of reproductive tissues (male hemipenes and female oviduct) and brain. Labeling for AR mRNA was very intense in the epithelium, but not within the body, of the male hemipenes. In contrast, expression of ER mRNA was prominent in most of the oviduct but not in the luminal epithelium. Within the brain, labeling for AR mRNA was conspicuous in the anterior olfactory nucleus, the lateral septum, the medial preoptic area, the periventricular preoptic area, the external nucleus of the amygdala, the anterior hypothalamus, the ventromedial hypothalamus, the premammillary nucleus, and the caudal portion of the periventricular nucleus of the hypothalamus. Expression of ER mRNA was sparse in the septum and was prominent in the ventromedial hypothalamus, the caudal portion of the periventricular nucleus of the hypothalamus, and a group of cells near the torus semicircularis. Many of these brain regions have been implicated in the regulation of hormone-dependent, sex-typical reproductive and agonistic behaviors in other vertebrates. Consequently, these nuclei are likely to control such behaviors in the leopard gecko and also are candidate neural substrates for mediating temperature effects on behavior. PMID- 11503142 TI - Guanine nucleotide exchange factors CalDAG-GEFI and CalDAG-GEFII are colocalized in striatal projection neurons. AB - CalDAG-GEFI and CalDAG-GEFII (identical to RasGRP) are novel, brain-enriched guanine nucleotide exchange factors (GEFs) that can be stimulated by calcium and diacylglycerol and that can activate small GTPases, including Ras and Rap1, molecules increasingly recognized as having signaling functions in neurons. Here, we show that CalDAG-GEFI and CalDAG-GEFII mRNAs, detected by in situ hybridization analysis, have sharply contrasting forebrain-predominant distributions in the mature brain: CalDAG-GEFI is expressed mainly in the striatum and olfactory structures and deep cortical layers, whereas CalDAG-GEFII is expressed widely in the forebrain. Within the striatum, however, the two CalDAG-GEF mRNAs have nearly identical distributions: they are coexpressed in striatal projection neurons that give rise to the direct and indirect pathways of the basal ganglia. Subcellular fractionation analysis of the substantia nigra with monoclonal antibodies against CalDAG-GEFI suggests that CalDAG-GEFI protein is present not only in the cell bodies of striatal projection neurons but also in their axons and axon terminals. These results suggest that the CalDAG-GEFs may be key intracellular regulators whereby calcium and diacylglycerol signals can regulate cellular functions through small GTPases in the basal ganglia circuits. PMID- 11503143 TI - Coupling from AII amacrine cells to ON cone bipolar cells is bidirectional. AB - The AII amacrine cell is a critical interneuron in the rod pathway of the mammalian retina. Rod signals pass into cone pathways by means of gap junctions between AII amacrine cells and ON cone bipolar cells. Filling AII amacrine cells with Neurobiotin produces labeling of cone bipolar cells by means of these gap junctions. However, tracer injections into bipolar cells do not produce labeling of the AII network (Vaney [1997] Invest Ophthalmol Vis Sci. 38:267-273), which suggests that the AII/bipolar gap junctions allow the passage of tracer in only one direction. This mechanism stands in contrast to physiological results, which indicate that light adapted signals can pass from ON cone bipolar cells into the AII network (Xin and Bloomfield [1999] Vis Neurosci. 16:653-665). Here, we report that a variety of ON and OFF bipolar cells are sometimes anomalously coupled to the A-type horizontal cell network. These relatively rare examples do not result from dye injection errors, but seem to represent minor developmental errors. However, this provides a method to obtain Neurobiotin-filled cone bipolar cells without the necessity of impaling them with a microelectrode. Under these conditions, Neurobiotin spreads from ON cone bipolar cells into neighboring AII amacrine cells. The dye-coupled AII amacrine cells, positively identified by double labeling with an antibody against calretinin, were centered around anomalously coupled ON bipolar cells. These results indicate that AII/bipolar cell gap junctions allow tracer coupling in both directions, consistent with previous physiological results. The previous failure to detect the passage of neuronal tracer from injected bipolar cells to AII amacrine cells may reflect electrode damage or perhaps the asymmetrical voltage sensitivity of a heterotypic gap junction. PMID- 11503144 TI - Morphology, electrophysiology, and calbindin immunoreactivity of myenteric neurons in the guinea pig distal colon. AB - The morphological and physiological characteristics of myenteric neurons in the guinea pig distal colon were determined using Lucifer yellow- or N-(2-aminoethyl) biotinamide-containing microelectrodes and intracellular recording and staining methods. The neurons in this study (n = 204) were classified on the basis of the shapes of their cell bodies and short processes or dendrites and the number of long processes or axons as Dogiel type I (n = 75 neurons; 36.8%), filamentous (n = 31 neurons; 15.2%), Dogiel type II (n = 38 neurons; 18.6%), and unclassified (n = 60 neurons; 29.4%). All Dogiel type II neurons had action potentials followed by an after-spike hyperpolarization (AH), and most of them (84%) had large, smooth somata and filamentous, short processes in addition to multiple, long processes or axons. Most of Dogiel type I, filamentous, and unclassified neurons (98%) had a single, long process, but four Dogiel type I neurons and one unclassified neuron had two long processes terminating as varicosities within other ganglia or on the surface of longitudinal muscle. The projections of monoaxonal neurons were distributed equally between oral and aboral directions, and most of them received fast excitatory postsynaptic potentials (EPSPs). All of the Dogiel type II neurons and seven Dogiel type I neurons were positive for calbindin immunoreactivity, but three filamentous neurons received fEPSPs, had spikes followed by AH, and were negative for calbindin. The presence of calbindin immunoreactive(-IR) neurons was quite variable among the ganglia. These results confirm that neither the presence of calbindin immunoreactivity nor the absence of fEPSPs can be used as a predictor of cellular morphology or electrophysiological properties of myenteric neurons in the distal colon. PMID- 11503145 TI - Number, origins, and chemical types of rat pallidostriatal projection neurons. AB - The dorsal globus pallidus (GP) receives major inputs from the dorsal neostriatum (Str), the subthalamic nucleus (STN) and the dorsal thalamus. The GP projects to multiple basal ganglia nuclei. One of the GP projection sites is the Str. The pallidostriatal projection has been considered minor. However, several recent studies have suggested that this projection is heavier than previously thought and that it might play a significant role in controlling the activity of the Str. To reveal more details of this projection, we examined the number of GP neurons that participated in the projection, their origins in the GP and their immunoreactivity for the calcium binding protein parvalbumin (PV), by using a combination of Fluoro-Gold (FG) retrograde labeling and immunohistochemical methods. Immunostaining for the calcium binding protein calbindin-28K (CaBP) was used to identify the CaBP-poor sensorimotor and CaBP-rich associative Str regions and the corresponding CaBP-poor middle, CaBP-rich border, and the caudomedial GP regions. The CaBP-poor dorsolateral Str region occupies a small portion of the Str, whereas the CaBP-poor middle GP region occupies a large portion of the GP. The immunostaining for neuron-specific nuclear protein (NeuN) was used to visualize neurons that were immunonegative for FG or PV. Cell counts revealed that the middle GP region contained a higher density of neurons and also a higher percentage of PV-positive neurons than the border and caudomedial regions of the GP. These observations suggested that the GP is involved more in sensorimotor function than associative function. Approximately 40% of neurons in the CaBP-poor middle GP region project to the CaBP-poor part of the dorsolateral Str. Approximately 30% of the neurons in both the CaBP-rich border and the caudomedial GP regions project to the CaBP-rich Str region. More than 40% of the pallidostriatal neurons in CaBP-poor middle GP region are PV-positive, whereas most of those in CaBP-rich GP regions are PV-negative. It was estimated from the cell count data that most of the PV-negative neurons in all three regions of the GP project to the Str. The results indicate that the sensorimotor and associative territories of the Str have reciprocal projections between corresponding territories of the GP. The involvement of a large number of GP neurons suggested that the pallidostriatal projection should be taken into account in the analysis of functional roles of the basal ganglia. PMID- 11503146 TI - Embryonic and paralarval development of the central nervous system of the loliginid squid Sepioteuthis lessoniana. AB - The embryonic development of the central nervous system (CNS) in the oval squid Sepioteuthis lessoniana is described. It has three distinct phases: (1) The ganglionic accumulation phase: Ganglionic cell clusters develop by ingression, migration, and accumulation of neuroblasts. (2) The lobe differentiation phase: Ganglia differentiate into lobes. The phase is identified by the beginning of an axogenesis. During this phase, neuropils are first formed in the suboesophageal mass, then in the basal lobe system, and finally in the inferior frontal lobes and the superior frontal-vertical lobe systems. (3) The neuropil increment phase: After the shape of the lobes reached its typical form, neuropil growth occurs, specifically in the vertical lobe. The paralarval central nervous system (CNS) is characterized by neuronal gigantism of the giant fibers and some suboesophageal commissures and connectives. The neuropil formation in the CNS of S. lessoniana occurs somewhat earlier than in Octopus vulgaris, although the principal developmental plan is quite conservative among the other coleoids investigated. Some phylogenetic aspects are discussed based on the similarities in the morphologic organization of their brains. PMID- 11503147 TI - Organisation and maturation of the human thalamus as revealed by CD15. AB - The distribution of the CD15 antigen (CD15, 3-fucosyl-N-acetyl-lactosamine, Lewis x) has been studied immunohistochemically in the fetal human thalamus. Its changing patterns could be related to three successive, but overlapping, periods primarily due to its association with radial glial cells, neuropil, and neural cell bodies, respectively. From 9 weeks of gestation (wg), a subset of CD15 positive radial glial cells distinguished the neuroepithelium of the ventral thalamus, a characteristic also seen in the developing mouse. Distal processes of the radial glial cells converged at the root of the forebrain choroid tenia, which was also CD15 positive. From 13 wg until approximately 20 wg, CD15-positive neuropil labeling marked the differentiation areas of prospective nuclei within the dorsal thalamus and progressively outlined their territories in a time sequence, which appeared specific for each nucleus. CD15 labeling of differentiating nuclei of the ventral, medial, anterior, and intralaminar thalamic divisions showed a transient topographic relationship with restricted areas of the ventricular wall. After 26 wg, CD15 immunoreactivity was observed in subpopulations of glial cells and neurons. Transient CD15 immunoreactivity was also found in delimited compartments within the subventricular region. The time of CD15 expression, its location, and cellular association suggest that CD15 is involved in segmentation of diencephalon, in the specification of differentiating nuclear areas and initial processes regarding the formation of intercellular contacts and cellular maturation. PMID- 11503148 TI - Exposure to fox odor inhibits cell proliferation in the hippocampus of adult rats via an adrenal hormone-dependent mechanism. AB - To determine whether exposure to fox odor alters granule neuron production, we examined proliferating cells and their progeny in the dentate gyrus of adult male rats exposed to trimethyl thiazoline, a component of fox feces. Additionally, to determine whether this effect is adrenal hormone-mediated, we examined animals exposed to fox odor after bilateral adrenalectomy and replacement with low levels of the endogenous glucocorticoid corticosterone. Stereologic analyses of the number of 5-bromo-2'deoxyuridine (BrdU) -labeled cells revealed that exposure to fox odor but not other, nonthreatening, odors (mint or orange) rapidly decreased the number of proliferating cells in the dentate gyrus. This effect is dependent on a stress-induced rise in adrenal hormones; exposure to fox odor resulted in an increase in circulating corticosterone levels and prevention of this increase (by means of adrenalectomy plus low-dose corticosterone replacement) eliminated the suppression of cell proliferation. Examination at longer survival times revealed that the decrease in the number of new granule cells in fox odor-exposed animals was transient; a difference was still detectable at 1 week after BrdU labeling but not at 3 weeks. In both fox and sham odor-exposed animals, many new cells acquired morphologic and biochemical characteristics of mature granule neurons. The majority of these cells expressed a marker of immature granule neurons (TuJ1) by 1 week after BrdU labeling and markers of mature granule neurons (calbindin, NeuN) by 3 weeks after labeling. These findings suggest that stressful experiences rapidly diminish cell proliferation by increasing adrenal hormone levels, resulting in a transient decrease in the number of adult-generated immature granule neurons. PMID- 11503149 TI - Development of GABA-immunoreactive neuron patterning in the spinal cord. AB - In the frog Xenopus laevis, gamma-aminobutyric acid (GABA)-immunoreactive spinal cord neurons (Kolmer-Agduhr cells) formed a dispersed pattern within two columns on either side of the midline. The cellular pattern became established during embryonic and larval development. The GABA-immunoreactive cells are cerebrospinal fluid (CSF)-contacting neurons that began to appear by 1.2 days (st 26) of development. This stage occurred shortly after neural tube closure (0.9 days, st 21) and followed the appearance of ultrastructural characteristics of CSF contacting neurons. The pattern of GABA-immunoreactive cells emerged during embryogenesis, as their density increased. Each longitudinal column was heterogeneous, containing cells with and without GABA immunoreactivity. Spatial analysis at several embryonic and larval stages showed that the cells in each column formed a nonrandom, dispersed pattern even at early stages of differentiation. This one-dimensional pattern resembled that of dopamine immunoreactive neurons, which are also located in the ventral spinal cord. The patterning of both cell types followed a different time course, but the ultimate spacing of the neurons remained comparable. These results suggested that the mechanism patterning the two cell types within the same region was similar but not identical and may involve related molecular mechanisms. PMID- 11503150 TI - The deep mesencephalic nucleus as an output center of basal ganglia: morphological and electrophysiological similarities with the substantia nigra. AB - The deep mesencephalic nucleus (DMN) is a large midbrain reticular region between the superior colliculus, the substantia nigra compacta, the periaqueductal gray, and the medial geniculate body. Although some data suggest that it is involved in nociception and visceral control, its functions remain unclear. In the present study, by using morphological (combination of anterograde and retrograde tracers with immunocytochemistry and in situ hibrydization) and electrophysiological (firing activity and transynaptic response to striatal stimulation) methods, we show that a subpopulation of DMN cells shares many morphological and electrophysiological characteristics with those of the substantia nigra reticulata (SNR). These similarities include the following: 1) firing rate, firing pattern, and conduction velocity; 2) expression of GAD65, GAD67, and PV; 3) excitatory and inhibitory inputs from the striatum; and 4) projections to the ventral thalamus, superior colliculus, and pedunculopontine tegmental nucleus. Some differences were also found. In comparison with SN, DMN cells and striatal afferents are more sparsely distributed and they show conspicuous contralateral projections to the thalamus and superior colliculus. This suggests that, similarly to the SNR, the DMN acts as an output center of basal ganglia and probably facilitates the inter-hemispheric regulation of these centers. PMID- 11503151 TI - Shape diversity among chick retina Muller cells and their postnatal differentiation. AB - It is currently believed that in each vertebrate species Muller cells in the central retina constitutes a fairly homogeneous population from the morphologic point of view and that particularly the chick Muller cell attains full shape differentiation at prenatal stages. However, in this study of the chick retina, from day 1 to day 55 of life, we show that there is a large variety of Muller cell shapes and that many of them complete shape differentiation postnatally. We used a cell dissociation method that preserves the whole shape of the Muller cells. Unstained living and unstained fixed cells were studied by phase-contrast microscopy, and fixed cells immunostained for intermediate filaments of the cytoskeleton were studied by fluorescence microscopy. Our results show that (1) Muller cell shapes vary in the origination of the hair of vitread processes, in the shape of the ventricular (outer or apical) process, in the presence or absence of an accessory process, as well as in the number and shape of processes leaving from the ventricular process at the level of the outer nuclear and outer plexiform layers (ONL/OPL); (2) during the first month of life, many Muller cells differentiate the portion of the ventricular process that traverses the ONL, most Muller cells differentiate the ONL/OPL processes, and all Muller cells differentiate the thin short lateral processes leaving from the vitread hair processes at the level of the inner plexiform layer (IPL). The number of cells differing in the shape of the ventricular process and that of cells with and without accessory process were estimated. The spatial relationship between the outer portion of the ventricular process of the Muller cell and the photoreceptor cells was also studied. Our results show that the branching of the ventricular process and the refinement of Muller cell shape is achieved without apparent participation of growth cones. We give a schematic view of how the branching of the ventricular process might take place and propose the size increase of photoreceptor soma as a factor responsible for this branching. PMID- 11503152 TI - Distribution of substance P and neurokinin-1 receptor immunoreactivity in the suprachiasmatic nuclei and intergeniculate leaflet of hamster, mouse, and rat. AB - The circadian pacemaker in the hypothalamic suprachiasmatic nuclei (SCN) receives photic information directly via the retinohypothalamic tract (RHT) and indirectly from retinally innervated cells in the thalamic intergeniculate leaflet (IGL) that project to the SCN. Using standard immunohistochemical methods, we examined the presence and distribution of substance P (SP) and the neurokinin-1 receptor (NK-1) in the SCN and IGL of rat and determined whether the patterns of immunostaining generalized to the SCN and IGL of Syrian hamster, Siberian hamster, and mouse. Terminals immunoreactive for SP were sparse within the SCN of Siberian and Syrian hamsters and mouse but were intense in the ventral, retinally innervated portion of the rat SCN. Immunostaining for the NK-1 receptor was mainly absent from the SCN of hamster and mouse. In contrast, a plexus of NK-1-ir cells and processes that was in close proximity to SP-ir terminals was found in the ventral SCN of the rat. Substance P-ir terminals were observed in the IGL of all four species, as were NK-1-ir cells and fibres. Double-labelled IGL sections of hamster or rat revealed SP-ir terminals in close apposition to NK-1 immunostained cells and/or fibres. These data indicate that SP could be a neurotransmitter of the RHT in rat, but not in hamster or in mouse, and they highlight potential species differences in the role of SP within the SCN circadian pacemaker. Such species differences do not appear to exist at the level of the IGL, where SP-ir and NK-1-ir were similar in all species studied. PMID- 11503153 TI - Connections of higher order visual relays in the thalamus: a study of corticothalamic pathways in cats. AB - Axonal markers injected into layers 5 and 6 of cortical areas 17, 18, or 19 labeled axons going to the lateral geniculate nucleus (LGN), the lateral part of the lateralis posterior nucleus (LPl), and pulvinar (P). Area 19 sends fine axons (type 1, Guillery [1966] J Comp Neurol 128:21-50) to LGN, LPl, and P, and thicker, type 2 axons to LPl and P. Areas 17 and 18 send type 1 axons to LGN, and a few type 1, but mainly type 2 axons to LPl and P. Type 1 and 2 axons from a single small cortical locus distribute to distinct, generally nonoverlapping parts of LP and P; type 1 axons have a broader distribution than type 2 axons. Type 2 axons, putative drivers of thalamic relay cells (Sherman and Guillery [1998] Proc Natl Acad Sci USA 95:7121-7126; Sherman and Guillery [2001] Exploring the thalamus. San Diego: Academic Press), supply small terminal arbors (100- to 200-microm diameter) in LPl and P, and then continue into the midbrain. Each thalamic type 2 arbor contains two terminal types. One, at the center of the arbor, is complex and multilobulated; the other, with a more peripheral distribution, is simpler and may contribute to adjacent arbors. Type 2 arbors from a single injection are scattered around and along "isocortical columns" in LPl, (i.e., columns that represent cells having connections to a common cortical locus). Evidence is presented that the connections and consequently the functional properties of cells in LP change along these isocortical columns. Type 2 driver afferents from a single cortical locus can, thus, be seen as representing functionally distinct, parallel pathways from cortex to thalamus. PMID- 11503154 TI - Connections of the nucleus incertus. AB - The nucleus incertus (NI) is a distinct cell group in caudoventral regions of the pontine periventricular gray, adjacent to the ventromedial border of the caudal dorsal tegmental nucleus. Recent interest in the NI stems from evidence that it represents one of the periventricular sites with the highest expression levels of mRNA encoding the type 1 corticotropin-releasing hormone (CRH) receptor, which has a high affinity for naturally occurring CRH, perhaps accounting for some of the extrapituitary actions of the peptide on autonomic and behavioral components of the stress response. However, almost nothing is known about NI function and hodological relationships. In this paper, we present the results of a systematic analysis of NI inputs and outputs using cholera toxin B subunit as a retrograde tracer and Phaseolus vulgaris-leucoagglutinin as an anterograde tracer. Our retrograde tracer experiments indicate that the NI is in a strategic position to integrate information related to behavioral planning (from the prefrontal cortex), lateral habenular processing, hippocampal function, and oculomotor control. Based on its efferent connections, the NI is in a position to exert significant modulating influences on prefrontal and hippocampal cortical activity, and the nucleus is also in a position to influence brain sites known to control locomotor behavior, attentive states, and learning processes. Overall, the present results support the idea that the NI is a distinct region of the pontine periventricular gray, and together with the superior central (median raphe) and interpeduncular nuclei the NI appears to form a midline behavior control network of the brainstem. PMID- 11503155 TI - Neuropsychological and functional cognitive skills of 35 unselected adults with sex chromosome abnormalities. AB - This report presents data defining the neuropsychological and cognitive phenotypes of a group of adults with sex chromosome abnormalities identified at birth through the chromosome screening of 40,000 consecutive newborns between 1964 and 1974. In all three nonmosaic groups, reading skills were impaired and intelligence quotients were on average reduced more than 20 points relative to controls. The 47,XXX women demonstrated greatest overall impairment, including reduced scores on tests of conceptualization and problem solving. 45,X women demonstrated impairment in spatial thinking skills, and 47,XXY men in verbal processing skills. No reduced scores were found in the female mosaic group. Marked variability in scores was seen in all groups; some propositi have been unable to hold any job, whereas others have completed college and are professionally employed. PMID- 11503156 TI - Identification of three novel mutations of the noggin gene in patients with fibrodysplasia ossificans progressiva. AB - We report noggin mutations in three Spanish families with fibrodysplasia ossificans progressiva (FOP). The three propositi have typical FOP findings; in the first and third families the parents are unaffected, while in the second family the father is partially affected. DNA of the three propositi and their parents was screened by sequencing for mutations in the noggin gene (NOG). Sequencing indicated a G to C mutation at nucleotide 274 of the NOG gene in the first propositus, encoding for the G92R substitution at the peptide level; this first mutation is de novo, the corresponding change not being observed in parents. In the second propositus, a G to T mutation at nucleotide 271 encodes for the G91C substitution, transmitted in the corresponding family by the partially affected father. In the third propositus, sequencing indicated a G to A mutation at nucleotide 275, encoding for the G92E substitution; this third mutation is de novo. All three mutations, as well as the Delta42 deletion already reported, resulted in the alteration of the portion of the NOG gene at positions 265-282, encoding for the potential N-myristoylation site at residues 89-GGGGGA 94. PMID- 11503157 TI - Phenotypic expression of tissue mosaicism in a 45,X/46,X,dicY(q11.2) female. AB - We describe a girl who presented at the age of 11 years with short stature. She had female external genitalia and some clinical features of Turner syndrome. At laparotomy a uterus and Fallopian tubes and small gonad-like tissue masses in the region of the Fallopian fimbria were found. The tissue masses were removed and histological examination revealed no organized testicular or ovarian morphology. Remnants of Fallopian tubes, epididymis, and clusters of Leydig cells were seen but no Sertoli cells were found. Endocrine studies showed levels of sex hormones consistent with primary gonadal failure. G-banding analysis of 16 blood lymphocytes revealed the karyotype 46,X,dicY(q11.2) in all cells. Varying proportions of X and Y centromeres in blood lymphocytes, skin fibroblasts, and in the incompletely formed Wolffian and Mullerian duct derivatives were demonstrated by FISH. Molecular studies confirmed the absence of most of the long arm of the Y chromosome and an intact short arm. The SRY gene was shown to be present, but we presume that due to the mosaicism the dose was insufficient to allow normal testicular development. PMID- 11503158 TI - Provisionally unique autosomal recessive syndrome due to significant consanguinity. AB - We report a patient with a syndrome of MR/MCA who was the product of a highly consanguineous family. The patient was the result of a union between a man and his daughter. The daughter was in turn the product of a mating between this same man and his mother. Major findings include: severe psychomotor and mental retardation, microcephaly with cerebral dysgenesis and cerebellar hypoplasia, early hypotonia and late hypertonia, short stature, early swallowing incoordination with aspiration pneumonias, distinctive face with striking hypertelorism, hypospadias, cryptorchidism, overlapping fingers, and rocker bottom feet. Chromosome and metabolic studies were normal. The severity and uniqueness of his phenotype suggests a new, probably private, autosomal recessive disorder due to homozygosity for one or more loci. PMID- 11503159 TI - Combination of renal agenesis with respiratory and alimentary tract atresia results in normal lung development. AB - The VACTERL complex comprises renal agenesis and atresias of the alimentary and respiratory tracts. We report on a case with this combination causing severe oligohydramnios but with normal lung development. The likely protective mechanism for pulmonary development was an increase in alveolar pressure and reduced alveolar fluid loss due to the esophageal-tracheal malformation. This suggests the possible treatment of oligohydramnios by tracheal occlusion. PMID- 11503160 TI - Soft tissue facial resemblance in families and syndrome-affected individuals. AB - We investigated soft tissue facial resemblance among relatives with or without syndromes and among related and unrelated individuals diagnosed with the same syndrome. Using correlation coefficients, we compared facial landmark (i.e., three-dimensional coordinate) positions and measurements gained by photogrammetry in various combinations of normal and syndrome-affected individuals. There were fewer significant correlations for the three-dimensional coordinates and measurements between the normal parent-normal child pairs than for the normal sib pairs. There was no discernible pattern for the single measurements in the parent child pairs, whereas all of the midline vertical measurements were significantly positively correlated in the normal sib pairs. Significant correlations were always positive in all sib comparisons, but ranged from negative to positive in all parent-child correlations. The shared environment of sibs was a possible explanation for their greater resemblance in comparison with parent-child pairs. We also had measurements from 11 subjects (related and unrelated) diagnosed with one of four syndromes, and we used these to compare individuals with the same syndrome by calculating correlation coefficients based on all available pairs of measurements. The highest significant positive correlations were found for related individuals with the same syndrome (0.72 to 0.83). Unrelated individuals with the same syndrome also had significant positive correlations, but they were lower (0.35 to 0.65). We therefore inferred that the genetic similarities between unrelated individuals with syndromes played a role in the resemblance between them, and that common genes and environment in related individuals further contributed to the high correlations found for them. PMID- 11503161 TI - De novo deletion of chromosome 18q in a baby with harlequin ichthyosis. AB - Harlequin ichthyosis, (MIM 242500), is a rare, autosomal recessive skin disorder due to an inborn error of epidermal keratinization. The gene for this condition has not been localized. We present a case of HI in which there was a de novo deletion of chromosome 18q: the karyotype was 46, XY, del(18)(q21.3). We postulate that the gene for HI may lie at, or distal to 18q21.3 and that the deletion observed in this case may have unmasked this autosomal recessive disorder. PMID- 11503162 TI - Heritability of life span in the Old Order Amish. AB - Although a familial contribution to human longevity is recognized, the nature of this contribution is largely unknown. We have examined the familial contribution to life span in the Old Order Amish (OOA) population of Lancaster County, Pennsylvania. Analyses were conducted on 1,655 individuals, representing all those born prior to 1890 and appearing in the most widely available genealogy, surviving until at least age 30 years, and with known date of death. Mean age at death (+/-SD) in this population was 70.7 +/- 15.6 years, and this did not change appreciably over time. Parental and offspring ages at death were significantly correlated, as were ages of death among siblings. Offspring longevity was correlated with longevity of both parents, and in more or less additive fashion. For example, mean offspring age at death was 69.4 +/- 15.3 years in individuals for whom both parents died before the age of 75 years (n = 280) and increased to 73.5 +/- 16.0 years in individuals for whom neither parent died before the age of 75 years (n = 311). These differences were highly significant (P = 0.006). We estimated heritability of life span to be 25% +/- 5%, suggesting that the additive effects of genes account for one quarter of the total variability in life span in the OOA. We conclude that longevity is moderately heritable in the OOA, that the genetic effects are additive, and that genetic influences on longevity are likely to be expressed across a broad range of ages. Published 2001 Wiley-Liss, Inc. PMID- 11503163 TI - A man who inherited his SRY gene and Leri-Weill dyschondrosteosis from his mother and neurofibromatosis type 1 from his father. AB - We report on a man with neurofibromatosis type 1 (NF1) and Leri-Weill dyschondrosteosis (LWD). His father had NF1. His mother had LWD plus additional findings of Turner syndrome (TS): high arched palate, bicuspid aortic valve, aortic stenosis, and premature ovarian failure. The proband's karyotype was 46,X,dic(X;Y)(p22.3;p11.32). Despite having almost the same genetic constitution as 47,XXY Klinefelter syndrome, he was normally virilized, although slight elevation of serum gonadotropins indicated gonadal dysfunction. His mother's karyotype was mosaic 45,X[17 cells]/46,X,dic(X;Y)(p22.3;p11.32)[3 cells].ish dic(X;Y)(DXZ1 +,DYZ1 + ). The dic(X;Y) chromosome was also positive for Y markers PABY, SRY, and DYZ5, but negative for SHOX. The dic(X;Y) chromosome was also positive for X markers DXZ1 and a sequence < 300 kb from PABX, suggesting that the deletion encompassed only pseudoautosomal sequences. Replication studies indicated that the normal X and the dic(X;Y) were randomly inactivated in the proband's lymphocytes. LWD in the proband and his mother was explained by SHOX haploinsufficiency. The mother's female phenotype was most likely due to 45,X mosaicism. This family segregating Mendelian and chromosomal disorders illustrates extreme sex chromosome variation compatible with normal male and female sexual differentiation. The case also highlights the importance of karyotyping for differentiating LWD and TS, especially in patients with findings such as premature ovarian failure or aortic abnormalities not associated with isolated SHOX haploinsufficiency. PMID- 11503164 TI - Novel and recurrent mutations in lamin A/C in patients with Emery-Dreifuss muscular dystrophy. AB - Emery-Dreifuss muscular dystrophy (EDMD) is characterized by slowly progressive muscle wasting and weakness; early contractures of the elbows, Achilles tendons, and spine; and cardiomyopathy associated with cardiac conduction defects. Clinically indistinguishable X-linked and autosomal forms of EDMD have been described. Mutations in the STA gene, encoding the nuclear envelope protein emerin, are responsible for X-linked EDMD, while mutations in the LMNA gene encoding lamins A and C by alternative splicing have been found in patients with autosomal dominant, autosomal recessive, and sporadic forms of EDMD. We report mutations in LMNA found in four familial and seven sporadic cases of EDMD, including seven novel mutations. Nine missense mutations and two small in-frame deletions were detected distributed throughout the gene. Most mutations (7/11) were detected within the LMNA exons encoding the central rod domain common to both lamins A/C. All of these missense mutations alter residues in the lamin A/C proteins conserved throughout evolution, implying an essential structural and/or functional role of these residues. One severely affected patient possesed two mutations, one specific to lamin A that may modify the phenotype of this patient. Mutations in LMNA were frequently identified among patients with sporadic and familial forms of EDMD. Further studies are needed to identify the factors modifying disease phenotype among patients harboring mutations within lamin A/C and to determine the effect of various mutations on lamin A/C structure and function. PMID- 11503165 TI - Prenatal diagnosis of a de novo ring chromosome 11. AB - Ring chromosomes are uncommon findings in prenatal diagnosis. Growth retardation is the most significant manifestation, in particular among patients with rings of larger chromosomes. A 30-year-old gravida 1, para 0 white woman was referred for genetic counseling because of maternal anxiety. Cytogenetic analysis of amniotic fluid cells at 16 weeks gestation revealed an abnormal mosaic female chromosome complement; 46,XX,r(11)(p15q25)[14]/45,XX,-11[7]. The ring 11 showed no detectable loss of chromosomal material at 450 band level. Both parents had a normal karyotype. Fluorescence in situ hybridization demonstrated intact subtelomeric regions in the ring chromosome. A targeted ultrasound evaluation at the time of consultation suggested no significant abnormalities. The parents were counseled and subsequently decided to terminate the pregnancy. The autopsy revealed an immature female fetus with abnormal craniofacial features including brachycephaly, low-set ears and hypertelorism, bicornuate uterus, and calcifications in the renal tubules. The abnormal phenotypes could be a consequence of the ring instability, submicroscopic deletion, and/or alteration of genetic material at the site of fusion. PMID- 11503166 TI - Preliminary evidence for a cognitive phenotype in Barth syndrome. AB - Barth syndrome (BTHS) is a rare, X-linked, recessive disorder that affects almost exclusively males. It is characterized by short stature, cardioskeletal myopathy, cyclic neutropenia, increased excretion of 3-methylglutaconic acid in the urine, and moderate hypocholesterolemia. The objective of the present study was to assess whether BTHS presents with a cognitive phenotype. Preliminary data were collected from five kindergarten or first-grade boys with BTHS. An abbreviated psychoeducational test battery was administered to each boy, and parents of each boy completed standardized behavior rating scales. Data from 120 boys of similar age or grade level were used for one comparison group; a subset of this sample comprised a comparison group that was individually matched on age and grade level to one of the five boys with BTHS. Preliminary data reflect a higher incidence of cognitive difficulties in boys with BTHS relative to both comparison groups. Boys with BTHS had significantly lower visual spatial skills, but comparable reading related skills, when compared with either group. Although based on a small sample size, the preliminary data presented in this work are the first indication of a cognitive phenotype associated with BTHS. PMID- 11503167 TI - Pure trisomy 10p resulting from an extra ring chromosome: characterization by methods of advanced molecular cytogenetics. AB - With the use of advanced molecular cytogenetic techniques, we have identified an extra ring chromosome consisting of the entire short arm of chromosome 10 (10p) in a 9-month-old girl with multiple congenital anomalies. This case represents another cytogenetic mechanism leading to the formation of pure complete trisomy 10p, which has not been reported previously. Pure trisomy 10p is rare. This case will further delineate those features associated with trisomy 10p. PMID- 11503168 TI - Frequency and ethnic distribution of the common DHCR7 mutation in Smith-Lemli Opitz syndrome. AB - Smith-Lemli-Opitz syndrome (SLOS) is an inherited multiple malformation syndrome caused by enzymatic deficiency of 3beta-hydroxysterol-Delta(7)-reductase (DHCR7). SLOS is thought to be most common among European Caucasians, with an incidence of 1 in 20,000 to 1 in 30,000 births. To define the carrier rate and ethnic distribution of SLOS, we screened DNA samples from 2,978 unrelated individuals for the most common SLOS mutation (IVS8-1G-->C). Twenty-four heterozygotes of the IVS8-1G-->C mutation were detected in 2,978 individuals of European Caucasian and Black backgrounds. For European Caucasians, the carrier rate for SLOS may be as high as 1 in 30, suggesting an incidence of 1 in 1,700 to 1 in 13,400. This high number is supported by the recent observation of newborn and prenatal incidence of 1 in 22,000 in the Caucasian population. Ours is the first report of the IVS8 1G-->C mutation in persons of African ancestry. Published 2001 Wiley-Liss, Inc. PMID- 11503169 TI - Rapid molecular prenatal diagnosis of Smith-Lemli-Opitz syndrome. PMID- 11503170 TI - Two novel frame shift mutations of CFTR causing null alleles in a patient with a severe course of CF. PMID- 11503171 TI - Lack of association of -308A/G TNFA promoter and 196R/M TNFR2 polymorphisms with disease severity in Thai adult malaria patients. PMID- 11503173 TI - [Computed tomography in the evaluation of the bronchopulmonary system in late periods after childhood partial resection of the lung]. AB - The authors used computed tomography (CT) as a basis to analyze the anatomic status of the bronchopulmonary system in 28 patients undergone resection of a part of the lung in childhood for chronic nonspecific lung inflammations and malformations. The findings confirmed the high diagnostic value of CT in the evaluation of the lung in different postoperative periods and showed a certain range of changes varying from significant in the lung operated on to progression of the primary disease to the collateral lung. PMID- 11503174 TI - [X-ray morphology of coronary arteries in patients with arterial hypertension in late periods after endovascular treatment]. AB - An association of arterial hypertension (AH) and late angiographic outcomes of endovascular interventions was studied in coronary heart disease. Forty two patients with or without AH underwent repeated coronary angiography within 2 months after the procedure. The total number of restenoses was found to be significantly greater in patients with AH than in those without it. The diameter of restenoses was significantly smaller (70.2 +/- 24.7 vs 48.2 +/- 29.2%) and their area is significantly larger (82.6 +/- 19.6 vs 66.9 +/- 28.8). Absence probability of restenosis was significantly smaller in patients with AH (7 vs 27%); at the same time there was the greatest reduction in this probability in the first 8 months following the intervention. PMID- 11503176 TI - [Radiodiagnosis of patients with primary sclerosing cholangitis]. AB - Among diseases of the hepatobiliary system, primary sclerosing cholangitis is an undetectable disorder of the biliary tract rather than a rare nosological entity, complex radiation study is of great importance in its preoperative diagnosis. Among direct methods for contrasting the biliary tract, the authors gave preference to percutaneous transhepatic cholangiography that allows the dilated biliary tract to be contrasted virtually in 100% of cases. The specific features of X-ray semiotics of primary sclerosing cholangitis were identified in 17 patients. PMID- 11503175 TI - [Possibilities of radiologic methods (ultrasonography, computed tomography) in the preoperative evaluation of intramural invasion of gastric cancer]. AB - By analyzing the findings in 72 patients with gastric cancer, the authors show the potentialities of noninvasive techniques of radiation diagnosis (transabdominal ultrasonography (USG) and X-ray computed tomography (CT) in the preoperative evaluation of the T-stage of gastric cancer. Ultrasound and computed tomographic semiotics of intramural invasion of gastric carcinoma is made. Ultrasonography was found to have the highest specificity in detecting early stage gastric cancers prior to computed tomography. In the authors' opinion, a complex use of the data obtained by these techniques reveals the degree of invasion of gastric carcinoma and its extent with high precision. Transabdominal USG and CT should rank with the initial methods used for diagnosing gastric cancer. Transabdominal USG should be most expediently used as an initial technique of the above studies, by taking into account its wide accessibility, easiness-to-use, and lack of radiation load. PMID- 11503177 TI - [Ultrasound study in the evaluation of the knee joint in osteoarthrosis deformans]. AB - The paper analyzes a group of 50 patients aged 25 to 55 years who had osteoarthrosis deformans of the knee joint. A control group comprised 10 healthy individuals of the same age. All the patients underwent routine roentgenography in 2 projections and ultrasonography of the knee joint by the procedure of L. Rubaltelli. The main criteria of sonographic destructive joint changes correlate with X-ray signs. The stage of the disease is defined by the magnitude of joint structural changes and by the sequence of their appearance, as shown by ultrasound study. PMID- 11503178 TI - [Intravascular embolization of cerebral aneurysms by mechanically detachable micro-spirals]. AB - The paper presents experience in embolizing arterial aneurysm, which of great value for interventional radiology. Endovascular embolization is a low-traumatic intervention, which reduces the time of rehabilitation in a patient and makes him active much earlier. The paper uses a sufficient body of data to provide evidence for the efficiency and safety of this technique in the treatment of aneurysms. The authors' experience allows one to expand indications for endovascular treatment of arterial aneurysm and to create a mechanism of embolization in practice, by applying the mechanically detachable spirals. PMID- 11503179 TI - [Systemic air embolism and pneumopericardium in a newborn with respiratory distress syndrome]. PMID- 11503180 TI - [Gated blood pool scintigraphy: diagnostic value of visual analysis]. PMID- 11503181 TI - [Radionuclide methods in the assessment of viability of dyssynergic myocardium in reversible left ventricular dysfunction]. PMID- 11503182 TI - [Effect of X-ray contrast media on hemostasis and its role in the genesis of adverse reactions]. PMID- 11503183 TI - [3D FSPGR (fast spoiled gradient echo) magnetic resonance imaging in the diagnosis of focal cortical dysplasia in children]. AB - Small dysplastic lesions of the cerebral cortex are often missed by conventional MRI methods. The identification of subtle structural abnormalities by traditional multiplanar rectilinear slices is often limited by the complex convolutional pattern of the brain. We used a method of FSPGR (fast spoiled gradient-echo) of three-dimensional MRI data that improves the anatomical display of the sulcal structure of the hemispheric convexities. It also reduces the asymmetric sampling of gray-white matter that may lead to false-positive results. We present 5 from 12 patients with dysplastic cortical lesions in whom conventional two-dimensional and three-dimensional MRI with multiplanar reformatting was initially considered normal. Subsequent studies using 3D FSPGR identified various types of focal cortical dysplasia in all. These results indicate that an increase in the detection of subtle focal dysplastic lesions may be accomplished when one improves the anatomical display of the brain sulcal structure by performing 3D FSPGR. PMID- 11503184 TI - [Stromal cell-derived factor 1 (SDF-1). Its structure and function]. AB - SDF-1, a novel cytokine from alpha-chemokine family, plays a key role in regulation of haematopoiesis. It exists in two forms (alpha and beta) that originate from alternative splicing. Its high expression in the bone marrow microenvironment accounts for the release of progenitor cells in the circulation and represents a prevention of uncontrolled leak of CD34+ cells. Notably significant is its stimulation of proliferation of B-lineage progenitors, in other haematopoietic lineages it functions as a facilitating factor of other cytokines. Ability of induction of platelet aggregation reveals the role of SDF-1 in thrombogenesis and vascular lumen obliteration in vessels affected by atherosclerosis. The only receptor for SDF-1 is CXCR4, whose presence was proved in great numbers of tissues and organs. Their presence was also verified in brain tumours, whereas degree of their expression raises with grading, angiogenesis and occurrence of necrotic changes in tumour. Thanks to this feature it will probably be possible to estimate the prognosis of the patients. SDF-1 is also a suppressor of immune response via its facilitating activity on the interaction of the macrophages and CD8+ T lymphocytes. Affinity of the T-lymphocytotropic HIV to CXCR4 holds out hopes for a possible modulation of the infection with SDF-1. The significance of SDF-1 and its receptor CXCR4 is supported by morphological and functional abnormalities of new-born mice in their absence, especially disorders in haematopoiesis, angiogenesis and development of cardiac and nervous tissues. PMID- 11503185 TI - [Histologic classification of kidney tumors for clinical practice in adults]. AB - Kidney tumors represent a wide scale of histological observations. However, only angiomyolipoma can be recognised preoperatively from results of the graphical examination. Other types can be recognised only on the bases of histological examination. Completely benign tumor is oncocytoma (it represents about 5% of all kidney tumors). Angiomyolipoma (2%) is also benign, though some case reports describing its malign transformation has been published. Angiomyolipoma under 4 cm can be only monitored, the larger tumors should be resected or selectively embolised the arterial blood supply to prevent spontaneous rupture. From the group of benign tumors only cystic nephroma can be diagnosed more often (up to 1%). One of the criteria for diagnosing the renal cortical adenoma is its size under 5 mm. That is why any adenoma, which could be diagnosed by means of graphical examination and therefore clinically significant does not exist. Most of tumors are malign epithelial tumors--renal carcinomas (RC). The are classified according Heidelberg classification into 5 elementary types: clear cell, papillary, chromophobe, originating form collecting ducts and not classifiable. Clear cell (conventional) renal carcinoma (CRC) comes most often (70 to 80%), its malign potential rise with increased size of tumor and with the gradient. Five year survival is achieved in 30-50%. Granular form of CRC carcinoma (7% of all CRC) is the equivalent of poorly differentiated PRC and it has an adverse prognosis. In contrary, the cystic form of CRC (about 6%) in benign. Papillary form of RC has the five-year survival in 84%, malignant are only tumors poorly differentiated. These are tumors with extensive necroses, which brings a fragile consistency and they can be distinguished by graphical examination. Chromophobe type of RC (5%) has the five-year survival in 90%. Poor prognosis has its sarcomatoid form, which can originate from any RC, but most frequently it is derived from the chromophobe type. The form originating from collecting ducts is highly infrequent and very malignant with the five-year survival in 20% only. The unclassified form of RC (3-5%) includes tumors not suiting to the criteria of the previous RC. Other primary renal malignant tumors (sarcomas, Wilms' tumor of adults, medullar carcinoma, carcinoid) are very rare. Comparatively frequent are metastases of other tumors (namely that of lung carcinoma) and renal impairment in leukemia, which are complication not often met by urologist. PMID- 11503186 TI - [Changes in microcirculation and selected laboratory parameters in the early stages of diabetic microangiopathy]. AB - BACKGROUND: Early stages of diabetic microangiopathy are accompanied with dysfunction, manifested by changes of some biochemical parameters. Parallel changes were observed in microcirculation. The aim of this study was to compare microcirculation in the skin of a forearm evaluated by laser Doppler with selected laboratory markers of endothelial dysfunction in Type 1 diabetes mellitus without microangiopathy or with incipient microangiopathy. METHODS AND RESULTS: Group of 43 Type 1 diabetic patients was examined in this study. 20 of them had no signs of microangiopathy and in 23 patients a simple diabetic retinopathy (background retinopathy) was diagnosed. Control group consisted of 25 healthy persons of comparable age, sex, and body mass index. All persons involved in this study were examined by laser Doppler and by biochemical examination and the results were compared. In comparison with control group, in diabetic patients the arm occlusion significantly lowered the increase of perfusion (29 +/- 12 vs. 41 +/- 18 perfusion units (PU) p < 0.01). Similarly the perfusion velocity increase was significantly lower in diabetic patients than in healthy controls (p < 0.01). Also the velocity of the perfusion increase after the warming up was lower in the diabetic than in non-diabetic persons (p < 0.01). Such changes of perfusion or those of velocity of perfusion increase were significantly lower in diabetic patients with microangiopathy than in those without this complication. Perfusion increases after both stimuli highly correlated (r = 0.86, p < 0.001). In diabetic patients with microangiopathy significantly higher N-acetyl-beta glucosaminidase (NAG) activities in serum and E-selection or ICAM-1 concentrations were found as compared with patients without microangiopathy, whereas plasma concentrations of tissue plasminogen activator (tPA) or inhibitor (PAI-1) were comparable with those in the control group. NAG activity inversely correlated with velocity of the perfusion increase after both the occlusion (r = 0.41, p < 0.01) and the warming (r = -0.38, p < 0.05). Similar relationship was found between tPA or E-selectin and the velocity of the perfusion increase after the occlusion (r = -0.48, p < 0.01). CONCLUSIONS: Our results confirm that biochemical parameters and microcirculation are impaired in the early stage of microangiopathy in Type 1 diabetic patients. Detailed analysis showed that both types of examination offer slightly different information on the vascular status. A long prospective study in diabetic patients without incipient vascular changes will be necessary to evaluate if biochemical or microcirculatory changes can bring an earlier information on the developing angiopathy. PMID- 11503187 TI - [Increased levels of circulating advanced glycation end products in a model of acute renal insufficiency in rats]. AB - BACKGROUND: Advanced glycation end products (AGEs) are formed from proteins and peptides by non-enzymatic glycation or glycooxidation. AGEs are formed slowly during aging, and they accumulate in circulation and tissues in diabetes and chronic renal failure. Kidney plays a key role in the disposal of AGEs. Aim of this study was to verify the hypothesis that, acute loss of renal function with enhanced oxidative and carbonyl stress should result in a rise of circulating AGEs levels. METHOD AND RESULTS: Acute renal failure (ARI) was induced in rats by bilateral nephrectomy (24-72 hours). The data on AGEs levels, oxidative status and antioxidative defense was compared to those of sham operated animals. 48 hours after the induction of ARI concentrations of AGEs, determined fluorimetrically or as carboxymethyllysine, rose 2-fold, and they correlated with concentrations of creatinine (r = 0.938, p < 0.001 and r = 0.815, p < 0.001, respectively). Malondialdehyde (MDA) and lipofuscine (LF) concentrations rose in a time dependent manner, suggesting an enhanced oxidative and carbonyl stress. Enhanced lipid peroxidation did not result from the suppressed antioxidant defense: activity of superoxide dismutase rose by 50%, while that of glutathione peroxidase was not compromised. Total antioxidant status increased, probably due to the accumulation of uremic toxins with scavenging capacity, such as hyppurate. CONCLUSIONS: According to our knowledge our data was first to show a rapid increase in circulating AGEs concentrations in the model of acute renal failure in rats. If AGEs accumulate in acute renal failure in humans, their contribution to acute toxicity, and/or to the development of later complications, might be of a great importance. PMID- 11503188 TI - [Ethylene glycol poisoning]. AB - Ethylene glycol is a common substance in various antifreeze preparations. Intoxication is caused by ingestion either as a suicide attempt or by an accident. The clinical course of intoxication is always severe, however, immediate launching of proper treatment can reverse the poor prognosis. Intoxication clinically proceeds in three stages: 1. Depressive CNS disturbances that can lead to coma. 2. Organ manifestations. 3. Acute renal failure. The case report of 46-year-old man admitted after ingestion of ethylene glycol in suicidal attempt summarises the essential principles of therapy--monitoring of vital functions, administration of ethanol solution as an antidote and hemodialysis to remove toxic components. PMID- 11503189 TI - [Glaucoma and pitfalls in therapy in the elderly with multiple morbidity from the viewpoint of the geriatrician]. AB - Glaucoma in multi-morbid patients is frequently treated by specialists non ophthalmologists. The growing number of elderly patients with glaucoma and comparatively small knowledge on this disease among the medical workers brought us to review recent findings on the aetiology, pathogenesis, risks of development and namely the problems of glaucoma treatment, possible drug interactions and their side effects. Considering the growing incidence of glaucoma with age, general practitioners, internists, geriatrists and other specialists will face the problems of drug interactions and global or ophthalmological side effects during the polypharmacological treatment of their multi-morbid seniors. The article gives basic information on the problems of using antiglaucoma drugs and their effects on the organism and eyes. Effects of some foodstuff and medicaments used in therapy of other diseases on the intraocular pressure and glaucoma are also summarise. PMID- 11503190 TI - [Current views on receptors for mediators of the autonomic nervous system of the heart]. AB - Heart function reflects activity of the heart pacemaker cells. This basic function is permanently influenced by two practically antagonistic systems- sympathetic and parasympathetic systems. They control not only the force of contraction (influencing calcium channels), heart rate (via potassium channels), but also arterioventricular transduction (changing the threshold for ion channels opening by influencing the maximal negative level of the membrane potential). Last few years changed our view on autonomic receptor expression in the heart. It is assumed that the heart adrenoreceptors are represented by alpha 1, beta, and beta 2 subtypes. The expression of beta 3 subtype is still controversial, similarly the existence of other, fourth beta-adrenoreceptor subtype, putative beta 4, is uncertain. Moreover, the end of the twentieth century brought the correction of former idea about a single muscarinic receptor subtype (M2) in the heart. Functional correlates of these findings are also reviewed. PMID- 11503191 TI - [Acute states in gastroenterology: spontaneous bacterial peritonitis and the acute intestinal pseudoobstruction syndrome]. AB - Our article concentrates on two acute states, which develop less dramatically but their after-effects may be very serious: Spontaneous bacterial peritonitis and Ogilvie's syndrome. Spontaneous bacterial peritonitis is a bacterial infection of the ascitic fluid without any intraperitoneal source of infection. Ascites is a condition of the disease but need not be clinically manifested. Spontaneous bacterial peritonitis comes usually during heavy hepatic impairment. Diagnosis can be set according: 1. Positive cultivation of ascitic fluid, 2. PMN levels higher than 250/mm3, 3. No infection, which may require a surgical intervention is apparent. Liver disease, which brings about the spontaneous bacterial peritonitis can be: 1. Chronic (e.g. alcoholic cirrhosis), 2. Subacute (e.g. alcoholic hepatitis), 3. Acute (e.g. fulminant hepatic failure). Mortality of this form of peritonitis can reach up to 46%. The most frequent etiological factor is alcohol and viral hepatitis, the most frequent agents are E. coli and Klebsiella pneumoniae. The disease is most effectively cured by cefalosporins of the third generation. With inadequate treatment, prognosis may be poor. Intestinal pseudoobstruction syndrome has clinical symptomatology of a serious impairment with ileus without signs of any mechanical intestinal obstruction. Syndrome can be classified according to its development: 1. Acute form--acute intestinal pseudoobstruction syndrome--Ogilvie's syndrome, 2. Chronic form- chronic intestinal pseudoobstruction syndrome. Pathogenic mechanism of the syndrome is not known. The disease is related to immobility, administration of some drugs, electrolyte imbalance and concomitant diseases (most frequently malignant tumors). Clinical symptomatology dominates nausea, vomiting, diffuse abdominal pain, constipation or diarrhoea. For diagnostics the first step should be termination of all medication, which could have causing affects, then taking native abdominal X-ray picture where gaseous intestinal distension can be prominent (coecum distended up to 9-12 cm). Identification of fluid surfaces is not usual. Endoscopic examination can exclude obstruction in the distal part of gut minimally. The most frequent complication is perforation of coecum. Pharmacological treatment relays on prokinetics. The basic intervention remains decompression by a rectal catheter or an effective coloscopic decompression with subsequent introduction of a cannula. Mortality of the disease fluctuates between 43 and 46%. PMID- 11503192 TI - [Activation of autologous hematopoietic stem cell transplants with interleukin 2]. AB - BACKGROUND: Recent findings of the role of the immunity in eradication of residual tumour tissue after autologous transplantation rejection leading to extensive studies on T-cell mediated specific antitumor effects or nonspecific NL cell mediated anticancer effects. We have evaluated on the methods of adoptive cell therapy--IL-2 activation of autologous graft in the preclinical conditions. In laboratory conditions we have manipulated with autologous grafts form patients suffering with chronic myelocytic leukemia and patients suffering with multiple myeloma. METHODS AND RESULTS: Autologous graft was activated with IL-2 during 24 hours cultivation period in X-Vivo 10 medium with heparine, glutamine and Dnase. Quality of grafts after cultivation, contamination and activation of T and NK cell were evaluated. No significant differences between IL-2 activated graft and control were found. Results of autologous graft quality (CD34+, CFU-GM) were comparable with already published results. Quality of final product allowed starting of clinical experimental trials. CONCLUSIONS: We have proved the possibility to use IL-2 activated autologous graft in the clinical conditions. Based on our preclinical results experimental clinical trials have been initiated in patients suffering from chronic myelocytic leukemia and multiple myeloma. PMID- 11503193 TI - [Pulmonary edema caused by laryngospasm]. AB - The authors report on a case of non-cardiac pulmonary oedema that developed in a young athletic man at an operating theatre. This unusual condition resulted from a laryngospasm followed by strenuous ventilation retractions. Diagnostic routine, differential diagnosis and treatment are described. PMID- 11503194 TI - [Health risks of mobile phones]. AB - Health problems related to the use of cellular telephones become a "hot" journalistic topic. Incompetent and uninformed authors of newspaper articles inform, deform and sometimes even frighten their users with malignant effects of electromagnetic radiation starting with impairment of psychic concentration, sleep disorders till development brain tumors. Such (and so far not proved apprehensions) have to be confound with scientific arguments. One of the first studies aimed at possibility of brain tumor formation brought about negative answer. The lack of empirical evidences of the mobile phone electromagnetic radiation cancerogenity suggests that cellular phones do not represent a serious risk of cancer. PMID- 11503195 TI - Storytelling is not just for children. PMID- 11503196 TI - Restringing instruments. PMID- 11503198 TI - Anterior cruciate ligament reconstruction. AB - The anterior cruciate ligament (ACL) is the most commonly reconstructed knee ligament. It is estimated that more than 100,000 new ACL injuries occur annually. This article discusses preoperative education, perioperative nursing care, surgical technique, and postoperative management of patients undergoing ACL reconstruction procedures. PMID- 11503199 TI - Thoracoscopic sympathectomy for palmar hyperhidrosis. AB - Palmar hyperhidrosis (i.e., excessive sweating of the palms) usually appears at puberty and causes psychological, social, educational, and occupational problems for people who suffer from it. Although many treatments have been used, the only treatment that permanently eradicates the condition is sympathectomy. The advent of thoracoscopic surgery has allowed surgeons to perform sympathectomy as an outpatient procedure that is safe and effective and produces life-changing results for patients. PMID- 11503200 TI - Sentinel lymph node biopsy for breast cancer. AB - In a health care system in which decreasing costs, increasing quality, and minimizing morbidity are emphasized, new, less invasive surgical techniques are gaining popularity. Sentinel lymph node biopsy is one such procedure used to treat many types of breast cancer. By performing a biopsy of this node, which receives lymphatic drainage from the tumor, the surgeon can predict the status of all nodes in the axillary region and accurately diagnose stages of cancer, thus eliminating axillary dissection for 55% to 70% of patients. Successful outcomes result from a concerted effort among the nuclear medicine physician, radiologist, surgeon, and surgical team members. PMID- 11503201 TI - The meaning of being a perioperative nurse. AB - The purpose of this study is to describe the meaning of being a perioperative nurse within the contexts of the sociopolitical, economic, and cultural forces that influence perioperative nursing practice. A theory of communicative action provided the framework for this philosophical inquiry. The researcher conducted one-time, in-depth interviews with six expert perioperative nurses selected from three practice settings in the northeastern United States. These interviews resulted in the text that the researcher analyzed based on the principles of critical hermeneutics, and four constitutive patterns were identified. The meaning of being a perioperative nurse for these nurses was identified as a struggle to make sense of their existence in the OR. PMID- 11503202 TI - Literature review and survey comparing surgical scrub techniques. PMID- 11503203 TI - Interviewing as a data collection method. PMID- 11503205 TI - Continuous free-flow electrophoresis separation of cytosolic proteins from the human colon carcinoma cell line LIM 1215: a non two-dimensional gel electrophoresis-based proteome analysis strategy. AB - The conventional approach for analyzing the protein complement of a genome involves the combination of two-dimensional gel electrophoresis (2-DE) and mass spectrometric based protein identification technologies. While 2-DE is a powerful separation technique, it is severely limited by the insolubility of certain classes of proteins (e.g. hydrophobic membrane proteins), as well as the amount of protein that can be processed. Here, we describe a simple procedure for resolving complex mixtures of proteins that involves a combination of free flow electrophoresis (FFE), a liquid-based isoelectric focussing (IEF) method, and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Resolved proteins were identified by peptide fragment sequencing using capillary column reversed-phase high performance liquid chromatography (RP-HPLC)/mass spectrometry (MS). An initial demonstration of the method was performed using digitonin/ethylenediaminetetraacetic acid EDTA extracted cytosolic proteins from the human colon carcinoma cell line, LIM 1215. Cytosolic proteins were separated by liquid-based IEF (pH range 3-10) into 96 fractions, and each FFE fraction was further fractionated by SDS-PAGE. Selected protein bands were excised from the SDS-PAGE gel, digested in situ with trypsin, and subsequently identified by on line RP-HPLC/electrospray-ionization ion trap MS. Our results indicate that FFE is: (i) an extremely powerful liquid-based IEF method for resolving proteins; (ii) not limited by the amount of sample that can be loaded onto the instrument; and (iii) capable of fractionating intact protein complexes (a potentially powerful tool for cell-mapping proteomics). An up-to-date list of cytosolic proteins from the human colorectal carcinoma cell line LIM 1215 can be found in the Joint Protein Structure Laboratory (JPSL) proteome database. This information will provide an invaluable resource for future proteomics-based biological studies of colon cancer. The JPSL proteome database can be accessed through the World Wide Web (WWW) network (http://www.ludwig.edu.au/jpsl/jpslhome.html). PMID- 11503206 TI - Thiourea enhances mapping of the proteome from murine white adipose tissue. AB - Adipose tissue imposes problems in two-dimensional (2-D) analysis due to its extremely high content of fat. To improve protein separation detergents and chaotropes were varied in the IEF step. The most important factor for obtaining distinct spots in the 2-D gel was whether thiourea was included or not. Many high molecular weight spots became resolved by using thiourea, while no spots disappeared or showed inferior characteristics, thus approximately twice as many spots were possible to quantify. Hydrophobic indices were compared for a set of proteins that gave rise to sharper spots with proteins that were not improved on the use of thiourea. The comparison did not give any statistically significant difference between the two groups of proteins. One of the effects obtained by inclusion of thiourea was that the dominating protein, serum albumin, appeared as more condensed spots allowing other minor proteins to be detected. This work resulted in a protocol which greatly enhances the resolution of proteins in adipose tissue. A 2-D map of mouse white adipose tissue from epididymal fat pads was constructed in which 140 spots were identified by mass spectrometry. This work lays the ground for our further studies on white adipose tissue in metabolic diseases such as obesity and dyslipidemia. PMID- 11503207 TI - Identification of protein carbonyls after two-dimensional electrophoresis. AB - The oxidative modification of proteins plays a major role in a number of human diseases, but identity of the specific proteins that are most susceptible to oxidation has posed a difficult problem. Protein carbonyls are increased after oxidative stress, and after derivatization with 2,4-dinitrophenyl hydrazine (DNP) they can be detected by various analytical and immunological methods. Although high resolution two-dimensional electrophoresis (2-DE) can resolve virtually all proteins present in a cell or tissue it has been difficult to determine the oxidized proteins because the DNP-derivatization process alters the isoelectric points of proteins, and additional procedures must be utilized to remove reaction byproducts. These additional procedures can lead to loss of sample, and poor isoelectric resolution on immobilized pH gradient (IPG) strips. We have developed a method that allows the IPG strips to be derivatized with DNP directly following isoelectric focusing of the proteins. This method allows the visualization of oxidized proteins by 2-DE with high reproducibility. PMID- 11503208 TI - A new silver staining apparatus and procedure for matrix-assisted laser desorption/ionization-time of flight analysis of proteins after two-dimensional electrophoresis. AB - We report on a new silver stain especially developed for staining large gels (25 cm x 20 cm) from the Hoefer ISO-DALT system for matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis of proteins. The staining protocol can be summarized as follows: the gels are sensitised in tetrathionate/potassium acetate solution and washed several times in distilled water. After impregnation with silver nitrate, the silver is reduced in the presence of potassium carbonate, thiosulphate and formaldehyde. The staining procedure is stopped with Tris/acetate after which the gels are rinsed and stored in water before spot picking for MALDI-TOF analysis is performed. This protocol has several advantages over existing ones. The gels are stained in a new apparatus that reduces gel handling to a minimum thus also reducing the contamination with keratins to a minimum. The development times in potassium carbonate are very long (up to 40 min) thus improving batch-to-batch reproducibility. Only the surface of the proteins is stained and the silver can be oxidized, thereafter MALDI-TOF can be performed with protein loads as little as 100 micrograms per gel. PMID- 11503209 TI - Rapid and simple single nanogram detection of glycoproteins in polyacrylamide gels and on electroblots. AB - The fluorescent hydrazide, Pro-Q Emerald 300 dye, may be conjugated to glycoproteins by a periodic acid Schiff's (PAS) mechanism. The glycols present in glycoproteins are initially oxidized to aldehydes using periodic acid. The dye then reacts with the aldehydes to generate a highly fluorescent conjugate. Reduction with sodium metabisulfite or sodium borohydride is not required to stabilize the conjugate. Though glycoprotein detection may be performed on transfer membranes, direct detection in gels avoids electroblotting and glycoproteins may be visualized within 2-4 h of electrophoresis. This is substantially more rapid than PAS labeling with digoxigenin hydrazide followed by detection with an antidigoxigenin antibody conjugate of alkaline phosphatase, or PAS labeling with biotin hydrazide followed by detection with horseradish peroxidase or alkaline phosphatase conjugates of streptavidin, which require more than eight hours to complete. Pro-Q Emerald 300 dye-labeled gels and blots may be poststained with SYPRO Ruby dye, allowing sequential two-color detection of glycosylated and nonglycosylated proteins. Both fluorophores are excited with mid range UV illumination. Pro-Q Emerald 300 dye maximally emits at 530 nm (green) while SYPRO Ruby dye maximally emits at 610 nm (red). As little as 300 pg of alpha 1-acid glycoprotein (40% carbohydrate) and 1 ng of glucose oxidase (12% carbohydrate) or avidin (7% carbohydrate) are detectable in gels after staining with Pro-Q Emerald 300 dye. Besides glycoproteins, as little as 2-4 ng of lipopolysaccharide is detectable in gels using Pro-Q Emerald 300 dye while 250 1000 ng is required for detection with conventional silver staining. Detection of glycoproteins may be achieved in sodium dodecyl sulfate-polyacrylamide gels, two dimensional gels and on polyvinylidene difluoride membranes. PMID- 11503210 TI - Multiresolution image registration for two-dimensional gel electrophoresis. AB - In proteomic research, two-dimensional electrophoresis (2-D) is an important tool for investigating differential patterns of qualitative and quantitative protein expression. The strength of the technique is due to its unrivalled power of being able to separate simultaneously thousands of proteins. The key to the comparison of 2-D protein profiles, however, lies in the use of a fast and robust image matching process which is essential to the subsequent quantification procedure. To satisfy the growing demand for a robust and fully automatic method of matching 2-D gel protein separation profiles, we describe in this paper a novel registration technique based on image intensity distribution rather than selected features. The method uses a multiresolution representation of the gel profiles and exploits the fact that coarse approximations to the optimal matching can be extracted efficiently from low-resolution images. This permits the removal of misalignments at different scales in a systematic manner and the strength of the new method has been confirmed by a double blind trial of 111 2-D gel pairs. The proposed method requires neither landmarks nor an a priori image alignment, and takes about five seconds for processing a typical gel pair on a standard personal computer. PMID- 11503211 TI - The use of glass wool as an attachment surface for studying phenotypic changes in Pseudomonas aeruginosa biofilms by two-dimensional gel electrophoresis. AB - Two-dimensional polyacrylamide gel electrophoresis was used to demonstrate phenotypic differences between Pseudomonas aeruginosa biofilm cells and the planktonic counterpart cells under defined culture conditions. Glass wool was used as a substratum for cell attachment as it affords a large surface-to-volume ratio (1 g with a mean diameter of 15 microns = 1300 cm2), supports the growth of biofilms, allows for free movement of cells between the inter-strand spaces, and it facilitates the exchange of nutrients and oxygen. It also allows for the separation of the biofilm biomass from the surrounding surface influenced planktonic (SIP) cells for further characterization. Comparative analysis of the respective proteomes indicated striking differences in the protein patterns of planktonic, biofilm and SIP cells. We selected 41 proteins, the levels of which varied in a significant and reproducible way in the respective protein profiles. In the biofilm cells, a general up-regulation of the spots was seen, but in SIP cells expression of these spots were generally down-regulated. Altogether six unique proteins were seen in the planktonic cells, while the biofilm and SIP cells contained five and two unique proteins, respectively. Glass wool, therefore, appears to be an ideal attachment surface for the study of biofilm development. PMID- 11503212 TI - Cellular distribution of translationally controlled tumor protein in rat and human testes. AB - In a recent proteomic study we identified 53 spermatogonial proteins among which was the translationally controlled tumor protein (TCTP). This is a protein previously reported as being implicated in proliferation events in normal and tumoral tissues that had never previously been seen in the testis. The present study was aimed at establishing the complete cellular distribution of TCTP and its transcript and the ontogenetic expression of this gene within the testis. Using an immunohistochemistry technique, an intense TCTP signal was detected in gonocytes (the prespermatogonia) in the fetal rat testis and in spermatogonia within adult human and neonatal and adult rat testes. Meiotic spermatocytes and postmeiotic haploid spermatids were also strongly immunostained in a stage dependent manner in human and rat testes. In addition, different levels of TCTP expression were also observed in the testicular somatic cells, with strong expression in Leydig cells and peritubular cells, and weak expression in Sertoli cells. Western and Northern blot analyses confirmed the presence of TCTP at all ages studied, with higher levels of RNA expression at 9 and 20 d postpartum, when spermatogonia and primary spermatocytes represent the highest proportion of germ cells: it was also confirmed that TCTP is present in all populations of isolated testicular cells. A transcript of 0.85 kb corresponding to TCTP, was expressed at all ages studied. This transcript was found to be expressed strongly in spermatogonia, somewhat less in isolated Leydig, resident macrophage, peritubular and Sertoli cells, weakly in the primary spermatocytes but not at all in spermatids. Interestingly, in the latter, a different transcript of 1.1 kb was present. The same 1.1 kb transcript appeared in testis extracts from 35 days postpartum onwards, corresponding to an age when spermatids accumulate within the tubules. Of note is that resident macrophages were found to express both the 0.85 and the 1.1 kb transcripts. We conclude that the strong expression of TCTP in spermatogonia makes it highly likely that the protein plays a significant role in spermatogenesis. PMID- 11503213 TI - Identification of tumor antigens in renal cell carcinoma by serological proteome analysis. AB - We have investigated the suitability of proteomics for identification of tumor associated antigens. First, we compared the proteomes of nontumorous kidney and renal cell carcinoma (RCC) by two-dimensional gel electrophoresis (2-DE) and silver staining. Protein patterns were markedly different (approximately 800 spots in RCCs versus approximately 1400 spots in kidney). 2-DE immunoblotting revealed five RCC-specific spots, reproducibly reactive with RCC-patient but not healthy donor control sera. Two of these antigens were isolated by preparative 2 DE, and identified by Edman sequencing of tryptic peptides. The first antigen, smooth muscle protein 22-alpha (SM22-alpha), is an actin-binding protein of unknown function predominantly expressed in smooth muscle cells. In situ hybridization revealed that SM22-alpha is not expressed in the malignant cells but in mesenchymal cells of the tumor stroma. The second antigen represents carbonic anhydrase I (CAI), an isoform usually not expressed in kidney. Interestingly, a different isoform (CAXII) has previously been identified by serological expression cloning as an antigen overexpressed in some RCCs. In additional assays, antibodies to recombinant CAI or SM22-alpha were detected in sera from 3/11 or 5/11 RCC patients, respectively, whereas sera from 13 healthy individuals did not react. In conclusion, serological proteome analysis may be a new tool for the identification of tumor-associated antigens. PMID- 11503214 TI - Proteins associated with the cell envelope of Trichoderma reesei: a proteomic approach. AB - A total of 220 cell envelope-associated proteins were successfully extracted and separated from Trichoderma reesei mycelia actively synthesizing and secreting proteins and from mycelia in which the secretion of proteins are low. Altogether 56 spots were examined by nanoelectrospray tandem mass spectrometry and amino acid sequence was obtained for 32 spots. From these, 20 spots were identified by Advanced BLAST searches against all databases available to BLAST. The most abundant protein in both types of mycelia was HEX1, the major protein in Woronin body, a structure unique to filamentous fungi. Other proteins identified were vacuolar protease A, enolase, glyceraldehyde-3-phosphate dehydrogenase, transaldolase, protein disulfide isomerase, mitochondrial outer membrane porin, diphosphate kinase and translation elongation factor beta. Partial short amino acid sequence obtained from some proteins did not allow them to be assigned to a specific protein in the database by BLAST search. In some cases, the tandem mass spectrometry spectra were too complicated to be able to assign an amino acid sequence with certainty. The number of spots (12) giving a clear signal but finding no match in the databases suggests that a majority of proteins associated with a filamentous fungal cell wall, are novel. Some technical problems related to protein isolation are also discussed. PMID- 11503215 TI - Let's put our shoulder to the problem of access to care. PMID- 11503216 TI - Taking up the challenge of injury control. PMID- 11503217 TI - Anal fissure: unclear causes but promising new treatments. PMID- 11503218 TI - Permanent cardiac pacing. PMID- 11503219 TI - Committing to change with an entry-level master's degree. PMID- 11503220 TI - Inserting an intrauterine device. PMID- 11503221 TI - Another pathway for generating compassion toward BPD patients. PMID- 11503222 TI - Borderline personality disorder in primary care. Recognizing signs and symptoms. PMID- 11503223 TI - Relationship management and functional improvement in the care of the BPD patient. PMID- 11503224 TI - Evidence-based medicine--in real time. Comparing methods of cervical Ca screening. PMID- 11503225 TI - Appendicitis in children. New insights into an age-old problem. PMID- 11503226 TI - Greeting a new time with new initiatives. PMID- 11503227 TI - [Sanitary and epidemiological services in chemical weapons elimination]. AB - Chemical weapons elimination on Russian Federation territory positions some problems that should be solved to prevent harm for public health and environment in Russia. The author specified corresponding methodic recommendations. PMID- 11503228 TI - [Silicosis hazard of silica dusts in diamond mining industry (experimental study)]. AB - Slow progressing experimental coniosis was induced by exposure to two samples of silica dust that was obtained from diamond openwork in "Mir" quarry of Yakutia. Moderate fibrogenicity of the dusts studied results from relatively low portions of silica and from metals oxides admixtures. PMID- 11503229 TI - [Interaction between cerebral hemispheres biopotentials under the influence of organic solvents (according to the spectral coherent analysis)]. AB - The study covered informative and diagnostic value of parameters in EEG spectral coherent analysis for early and differential diagnosis of chronic exposure to organic solvents. Neurotic symptoms appeared to be early signs of the toxic effect and to be associated with dramatic decrease in regulatory influence from thalamic and hypothalamic structures. PMID- 11503230 TI - [Diagnostic problems of post-intoxication states]. AB - Complete clinical examination covered 37 patients in various stages after acute occupational poisonings with pesticides (8 examinees), with explosive gases (9 subjects), with carbon oxide (3 examinees), with cadmium oxides (1 examinee), with nitrogen compounds (4 ones), with sulfurous gases (2 examinees), with organic solvents (10 subjects) including chlorinated hydrocarbons (2 subjects). Post-intoxication period appeared to include syndromes characteristic for each poison and corresponding to severity of acute state (from cephalgia to parkinsonism and thyrotoxicosis after acute poisoning with carbon oxide, organic neurologic signs and toxic hepatitis after acute poisoning with phosphorus organic compounds, cerebral asthenia after acute exposure to organic solvents, severe encephalopathy and toxic auditory and optic neuropathy after hydrogen sulfide). Inadequate medical rehabilitation and continuous occupational exposure induced deterioration in clinical signs and advanced post-intoxication syndromes. PMID- 11503231 TI - [Hygienic regulation of welding aerosols in air of workplace and pneumoconiosis in welders (review of literature)]. AB - The authors represent review of literature on fibrogenic effects of welding aerosols and results of own clinical and epidemiologic studies concerning pneumoconiosis course in electric welders compared to other occupational groups. In conclusion, the authors necessitate more strict control over total concentration of welding aerosol solid component (WASC) in air of workplace and determination of MAC for WASC. PMID- 11503232 TI - [Gas chromatography in detection of toluene diisocyanate in biological medium]. AB - The authors suggested a method of gas chromatography for detection of toluylene diisocyanate (TDI) in biologic material: liver, lungs, spleen, brain, kidney. The method includes TDI extraction from tissues by solvent, subsequent concentration and the extract chromatography by a device with electron capture detector. The method was tested in white rats. PMID- 11503233 TI - [Reactivity of bronchi in workers exposed to various pneumotropic substances]. AB - Determining risks of bronchial asthma is one of the main tasks in contemporary occupational pulmonology. The important risk factor is nonspecific increased reactivity of bronchi. The work was aimed to determine occurrence of nonspecific increased reactivity of bronchi in individuals working with pneumotropic polluters and to test a new method determining. PMID- 11503234 TI - [State of respiration biomechanics and blood gases in electric welders having various occupational diseases]. PMID- 11503235 TI - [Occupational hygiene in building ceramics production]. PMID- 11503236 TI - [Changes in biochemical, cytochemical and immunologic parameters under exposure to low levels of nickel in humans]. PMID- 11503237 TI - [Hereditary polymorphism features in patients with silicosis associated with chronic bronchitis]. AB - Chronic bronchitis frequently accompanies silicosis in metallurgy workers. Association of the two diseases significantly lowers life quality and disables the patients. Prophylactic measures for the diseases are elimination of the etiologic factors and detection of genetic markers for propensity and insusceptibility to silicosis and chronic bronchitis. Phenotypes of Lewis a-b- system, of Hp2-1 haptoglobin and of group-specific component Gc2-2 could be risk factors for chronic bronchitis in silicosis patients. Those of Lewis a-b+ system, of Hp2-2 haptoglobin and group-specific component Gc1-2 are resistant to chronic bronchitis on silicosis background. PMID- 11503238 TI - Treating subacromial bursitis with an injection into the bursa. PMID- 11503239 TI - Current practices and newer agents for the management of pediatric asthma. PMID- 11503240 TI - Detox in the ED: taking urgent action. PMID- 11503241 TI - Asthma and impotence. The story of an unexpected connection. PMID- 11503242 TI - Rectifying color and class disparities in health care. PMID- 11503243 TI - Responding to evidence of abuse in the elderly. PMID- 11503244 TI - Football or cervical Ca?--make up your mind. PMID- 11503245 TI - Answers to century-sized questions. PMID- 11503246 TI - Answers to century-sized questions. PMID- 11503247 TI - The culturally sensitive medical interview. PMID- 11503248 TI - [Molecular basis of carcinogenesis in lung cancer induced by cigarette smoking]. AB - An association between cigarette smoking and lung cancer carcinogenesis is reviewed. It is highly possible, that "individual susceptibility" for tumor development exists and is related to polymorphic variants of genes encoding for enzymes, which are employed in metabolism of xenobiotic substances. The gathering of highly reactive molecules due to modified metabolic processes results in DNA adducts forming and increased tendency for mutations. Group of genes, responsible for proliferation, cell cycle arrest, apoptosis and DNA damage repair are frequently altered. PMID- 11503249 TI - [Cytological assessment of the nasal mucosa in children with adenoidal hypertrophy. Part I]. AB - Investigation was performed on 68 children (24 girls and 44 boys) aged of 3-14, who was qualified by paediatricians for cytological evaluation of the nasal mucosa. They had undergone treatment for chronic upper respiratory tract infection and had clinical symptoms of the adenoidal hypertrophy. The adenoidal hypertrophy in children was confirmed by x-ray of nasopharynx. Seven children had previously undergone adenoidectomy and 3 had recurrent otitis. In the investigated group of children the majority of cytograms contained high level of neutrophils (> 50% of the total number of cells). Only 2 children showed normal cytogram. Discovery of substantial eosinophilia of the nasal mucosa (11.76% of investigated children) indicates that allergy diagnosis should be performed in children with adenoidal hypertrophy. PMID- 11503250 TI - [Cytological assessment of nasal mucosa in children with adenoidal hypertrophy. Part II]. AB - In this investigation we would like to evaluate what changes in cytology of the nasal mucosa occur after surgical removal of adenoids. The investigation was performed on 25 children (10 girls and 15 boys) aged 3-13. Material for the cytological evaluation was obtained from the nasal concha. The children were tested twice: before and one month after the surgery. Cytograms of the nasal mucosa of children with adenoidal hypertrophy before the surgery showed increased number of neutrophils. Cytological normalisation of the nasal mucosa occurs in one month after the surgical removal. In two children percentage of eosinophils in cytograms increased and percentage of neutrophils remained significant after the surgery. Cytological assessment of the nasal mucosa performed before and after surgical removal of the hypertrophied adenoid is helpful in evaluation of the accomplished therapy and may indicate what are the reasons of its failure. PMID- 11503251 TI - [Selenium plasma concentration level in children with food allergy]. AB - Selenium (Se) is an essential component of many mammalian enzymes and therefore plays an important role in different metabolic processes. Se deficiency occurs in inadequate diet consumption as well as in some digestive tract and allergic diseases. In this research we showed that Se plasma concentration levels in healthy children was 71.8 mg/l, whereas in those with food allergy 54.1 mg/l (with smaller intestinal villus atrophy) or 50.4 mg/l (with greater intestinal villus atrophy). Obtained results indicate that children with food allergy display higher risk of Se deficiency. Antioxidative and immunomodulatory action of Se, connecting with promising literature data of Se supplementation, promote us to state, that this trace element could be used for accessory food allergies treatment. PMID- 11503252 TI - [Cigarette smoking as a predicting factor for osteoporosis in women]. AB - The aim of the study was to answer the question--if smoking could facilitate occurrence of osteoporosis in chosen population?. Study was among 110 women in the age between 45-66 years, hospitalised in the Hospital in Slupsk. In 22 (20%) of respondents osteoporosis was diagnosed (bone density was lower than medium density for more than 2.5 SD in densytometric study), and these women were qualified to group I. Group II consisted of 88 (80%) women who were not diagnosed towards osteoporosis. Based on this study, it can be stated, that in examined group smoking could facilitate occurrence of osteoporosis among respondents from group I, what is proved by a high percentage of smokers 68.2%, the average number of cigarettes smoked per day 17.0 +/- 1.2 items and average period of smoking 32.1 +/- 6.5 years. As smoking is common phenomenon among women (one out of two examined smoked) you should stress elimination of this risk factor of many diseases including osteoporosis. PMID- 11503253 TI - [Own experience in diagnosis and therapy in pulmonary Langerhans cell granulomatosis]. AB - We have assessed 12 patients (2 females, 10 males) aged between 19 and 53 years (mean 38.3 +/- 10.3) diagnosed with LCG during 14-year period (1985-1998). All patients were smokers. Follow-up was from 6 to 132 months (mean 47.5 +/- 44.4). LCG diagnosis was confirmed by histology in 10 cases (9 lung, and 1 bone biopsy). In 2 patients the diagnosis was made on clinical grounds, including characteristic appearance on HRCT scans. Mean FVC was 78.9 +/- 15.9% of predicted, DLCO 64.1 +/- 22% of predicted. In 8 patients (67%) FVC or DLCO were below 80% of predicted. In 2 patients with histologically proven diagnosis, HRCT was not characteristic for LCG. The treatment was introduced in 8 patients (67%). Only 3 out of 8 patients initially treated with steroids responded to this treatment. In the rest treated patients, therapy was changed to cytotoxic agents. 3 patients died (1 after 2 years, and 2 after 11 years) 2 due to LCG and 1 due to pneumonia. CONCLUSION: HRCT appearance is not always characteristic in patients with histological diagnosis of LCG. Pulmonary involvement in LCG is frequently connected with lung function derangements. Response to steroids is poor, and switching to cytotoxic agents is often necessary. PMID- 11503254 TI - [Evaluation of anti-mycobacterial antibody IgG concentration in rapid diagnosis of tuberculosis]. AB - Tuberculosis is not always easy to diagnose. All the methods applied in diagnosis of this illness have certain limitations. In this study the value of Rapid Test TB in diagnosis of tuberculosis was evaluated. The aim of the test is to detect antibody against mycobacterial antigen 38 kDa. Material consisted of 100 persons divided into 4 groups: BK(+) tuberculosis patients, BK (0) tuberculosis patients- culture (+), lung cancer patients and patients with sarcoidosis. Positive results were obtained in 100% of culture positive patients and in 10% of patients culture (+), in 20% of lung cancer patients and in 40% of patients with sarcoidosis. Specifity of the test was 55%. The results indicate that Immunozyme Mycobacterium test is valuable tool in tuberculosis diagnosis especially in case when a technical equipment is wanting. PMID- 11503255 TI - [Endothelin-1 in pathogenesis of Raynaud's syndrome]. AB - Endothelin is an endogenous vasoconstrictor and plays an important role in pathogenesis of Raynaud's phenomenon. Plasma endothelin-1 (ET-1) and von Willebrand factor (vWF) concentrations following cold exposure in 52 patients with Raynaud's phenomenon were measured. Statistically significant increase of ET 1 and vWF was found in the study group in compare to healthy volunteers. There was positive correlation between ET-1 and vWF in those cases. The dates suggest that ET-changes indicates a vasospastic effect on vascular injury. Treatment with endothelin-receptor antagonist may prevent structural changes in vessel well. PMID- 11503256 TI - [Congenital pneumonia in fetuses and newborns ad inflammatory lesions in the placenta]. AB - The authors compared the degree of congenital pneumonia in stillborn and neonates died in first two days of life with inflammation lesions of placenta. The coexistence of those two processes and its significant correlation was found, especially among preterm LBW neonates. This finding supports the hypothesis about infection as a probable cause not only of preterm deliveries but infection of fetuses and neonates as well. Analysis of some cases revealed also that general infection of mother could cause transplacental infection of foetuses. PMID- 11503257 TI - [Circulatory and respiratory systems in children after surgical correction of atrial septal defect type II (ASD II)]. AB - The main objective of the study was to evaluate the conditions of circulatory and respiratory system 1-5 years after surgical correction of atrial septal defect (ASD II) in 112 children who had been operated at Silesian Medical Centre in Katowice in 1989-1993. Obtained results were compared with these ones found in the group of 60 healthy children. Mean age of study group was 7.78 +/- 4.46 years at the time of examination and 5.34 +/- 3.98 at the time of the operation. Twenty older children in the mean age 11 +/- 4.5 years (necessity for cooperation) were selected from the study group to perform the respiratory system tests. These tests were performed 7 days prior to operation and repeated 8 to 12 months afterwards. The following indices were determined: FVC--forced volume capacity, FRC--functional residual capacity, RV--residual volume, TLC--total lung capacity, FEV1--forced first second expiratory volume, PEF--peak expiratory flow, MEF 75-50 25% FVC--maximum expiratory flow at 75-50-25% of forced volume capacity, ITGV- intrathoracic gas volume, Raw-respiratory tract resistance, CL, st--static compliance of lung, DL, CO[SB]--lung diffusion capacity for carbon monoxide and D/VA--Krogh transfer coefficient. Circulatory system tests included: medical interview, physical examination, routine and 24-hour ambulatory ECG, echocardiography. There was found that children after ASD II closure have efficient circulatory system and most of them have normal ECHO results. Almost 40% of the children within 1 to 5 years following ASD operations have cardiac arrhythmias found in 24-hour ECG, but significant arrhythmias are seen only in 17.8% of the subjects. Most of the rhythm disturbances were mild. Significant arrhythmias and sinus node dysfunction were observed more frequently among the children who underwent surgical correction than in the healthy control group. Statistically significant deterioration of lung diffusion capacity for carbon monoxide (DL,CO[SB]) and Krogh coefficient (D/VA) has been found after the surgery. Thus, it is concluded that some pulmonary abnormalities may persist or even increase after successful ASD operation. PMID- 11503258 TI - [Virtual bronchoscopy: a new tool of non-invasive imaging of airway inner surface based on standard spiral computed tomography of the chest]. AB - The aim of the study is to assess the new method of radiological imaging of the inner surface of a tracheobronchial tree in order to determine to what extend it can replace the standard bronchoscopy. The spiral CT examinations of 43 patients with hilar lung tumor in virtual mode had been reconstructed retrospectively and compared to the results of fiberoptic bronchoscopy. There was a significant correlation between both methods to reveal stenoses or occlusions of airway lumen. Virtual endoscopy turned out to be an unsatisfactory method in the visualisation of mucosal changes. PMID- 11503259 TI - [Drug resistance of group B streptococci]. AB - The group B streptococci (GBS) are one of the main infections' factors in neonates. The source of the infection is mother and the infection follows during childbirth. In many countries various methods of prophylaxis in reference to mother and neonate are introduced. Thus far the widest application have found a method of giving to pregnant women penicillins or cephalosporins. The purpose of the present work was to study how the drug-resistant of the GBS forms in our country and if using antibiotics in prophylactic purposes without doing any antibiograms is burdened with risk, resulted from the possibility of meeting resistant strains. 365 strains of the GBS, derived from colonised neonates, pregnant women and clinical materials were put to the scientific research. 120 animal strains isolated from events of mastitis in cows, represented the second group. The GBS, derived from pregnant women were sensitive to every examined antibiotics except lincomycin (5.9%). Amid the streptococcus isolated from neonates there were some strains resistant to every examined antibiotic (1.9 4.4%). More often resistant strains were isolated from clinical materials (3.9 9.0%). The highest percentage of the resistant strains was identified in animals (7.5-20.0%). The GBS as a rule were insensitive to cotrimoxasolum. As follows from our scientific researches, the GBS derived from people as a rule are sensitive to used antibiotics. But there were some strains resistant even to several antibiotics. Possibility of meeting these strains should be taken into consideration as in prophylaxis as in treatment of the infections. Amid the animal strains of the GBS the phenomenon of the resistance to antibiotics is more often met than in human ones. PMID- 11503260 TI - [Patient adherence in respiratory tract infections: ceftibuten versus other antibiotics (PARTICULAR study)]. AB - The study was conducted to evaluate patient compliance to antibiotic therapy in respiratory tract infections. The main aim of the study was to establish whether dosing frequency (1 vs 2 or 3 times daily) and other factors influence compliance. Patients aged 18 and over attending selected primary health facilities in Lodz region were enrolled. Patients were randomly assigned to receive ceftibuten, 400 mg once daily or other antibiotic on physician's choice with 2 or 3 times daily dosing. On the 5th day during the home visit the questionnaire study was performed and the compliance was measured by a pill count. Four hundred and six patients (205 in ceftibuten and 201 in other antibiotics group, respectively) were fully evaluable. Overall compliance was 76.6% with 97.6% for ceftibuten, 66.0% for antibiotics with b.i.d. dosing and 23.5% for antibiotics with t.i.d. dosing. Using a logistic regression analysis with a stepwise variable selection, dosing frequency was found to be a major variable associated with patient compliance (p = 0.00000, odds ratio 0.09, 95% confidence interval 0.057-0.165). Non-compliance with antibiotics is a common phenomenon in respiratory tract infections therapy. Once-daily antibiotic dosing leads to the best possible compliance in such circumstances. Due to its once daily dosing and rare side effects, ceftibuten ensures nearly perfect compliance in respiratory tract infections therapy. PMID- 11503261 TI - [Immunological marker activity in first episode schizophrenic patients]. AB - We evaluated immune-inflammatory markers in 32 first-episode schizophrenic patients during exacerbation of symptoms and during clinical improvement. Mean concentrations of sIL-2R and IFN-g was higher and mean concentration of sIL-6R was lower in cell cultures from blood of schizophrenic patients than in normal controls and there was no difference in IL-2 concentration. There was no correlation between concentrations of cytokines and demographic and clinical data. Our results may suggest that immune system disturbances could be observed in schizophrenia at the onset of the disease. PMID- 11503262 TI - [Hoigne syndrome as an acute non-allergic reaction to different drugs: case reports]. AB - In 1959 Hoigne described the first cases of pseudo-anaphylactic reactions induced by intramuscular administration of procaine penicillin G. This complication, characterized by acute psychological and neurological manifestations, is still of current interest. We report cases of patient, whose Hoigne syndrome was not due to procaine penicillin G (i.e. lidocaine, edan, azlocillin, gentamycin, klaritromycin and cefuroxime). It is accepted that Hoigne syndrome can be caused by lidocaine. Further Stell i.m. and Ojo OA published similar case report of Amoxycillin-induced hallucinations. These clinical observations indicate also that using term "acute non-allergic reaction to procaine penicillin G" as synonym of Hoigne syndrome is not precise. Further classification of the reaction among psychic disturbances or adverse drug reactions is very difficult. Much of literature is devoted to the differentiation from anaphylaxis rather than other symptoms of main disease or side effects. Harmful effect of main disease is very likely. Lidocaine was taken intravenously by patient with diabetes and heart attack or locally with chronic hepatitis during topical anesthesia. Benzylpenicillin, azlocycillin, gentamycin, klaritromycin and cefuroxime was used during serious bacterial infections. These observations indicate that pathogenesis of Hoigne syndrome may be associated with main disease and organic brain disease. Then procaine kindling mechanism partly explain Hoigne syndrome. Therefore pathogenesis, definition, differentiation and classification of this complication is still unclear. PMID- 11503263 TI - [Cutaneous cryptococcosis in AIDS patient]. AB - 45-year-old man with the Acquired Immunodeficiency Syndrome, on highly active antiretroviral therapy (HAART) resulting in rapid decline of HIV RNA and increase of CD4 T cells count, developed multiple skin umbilicated lesions (resembling molluscum contagiosum) on his face, ears, neck and chest. Histopathology and mycological cultures of a skin biopsy revealed Cryptococcus neoformans. Antigens of Cryptococcus was also identified in blood. During treatment with amphotericin B, the skin lesions regressed. This case demonstrates that skin lesions resembling molluscum contagiosum may be caused by cryptococcal infection. It is necessary to perform skin biopsy in HIV-infected persons with skin lesions to diagnose cutaneous cryptococcosis. The open question is if skin cryptococcosis may be the immune reconstruction disease. PMID- 11503264 TI - [A case of primary lymphangiomyomatosis in retroperitoneal space and lungs]. AB - Authors present a rare case of primary lymphangiomyomatosis. Diagnosis was based on histopathological evaluation. Diagnostic difficulties were caused by lack of pathognomonic symptoms in clinical examination, ultrasonography and CT. The patient has been treated with hormonal medicines (Provera, Megace) since 1997. Now she is in a good condition, She hasn't got any symptoms of progression disease. PMID- 11503265 TI - [Liver tuberculosis]. AB - The case of rare localization of extrapulmonary tuberculosis--isolated tuberculoma of the liver--was described. The tumor was found accidentally, during abdominal ultrasound for benign prostatic enlargement. Computed enhanced tomography didn't explain the character of the tumor, so percutaneous thin-needle biopsy was performed. Preliminary histological report suggested primary liver cancer. Patient was operated on, tumor was removed with the margin of the healthy liver tissue. Histological examination of the specimen revealed the nature of the tumor. Diagnosed liver tuberculoma is usually treated without surgery. PMID- 11503266 TI - [Adhesion molecules and their role in organ response after trauma]. AB - In this paper, we presented general characterization of adhesion molecules and their role in organ response after injury. Severe trauma is frequently complicated by multiple organ dysfunction (MODS) or failure (MOF). The pathogenesis of these syndromes involves a complex interaction of humoral and cellular events. The recruitment of circulatory leukocytes into tissues is regulated in part by specific leukocyte-endothelial cell interactions with several families of cell adhesion molecules. In the adhesion cascade, the initial phase of inflammation, a transient slowing of neutrophils in postcapillary venules is mediated by selectins. Firm adhesion of neutrophils to the endothelium occurs by interaction of beta 2 integrins to ICAM-1. Diapedesis between endothelial cells requires the engagement of PECAM-1. Significance of the adhesion molecule in organ response after severe trauma permitted their diagnostic and prognostic use as well as new methods of treatment. PMID- 11503267 TI - [Allergy to penicillin: facts and controversies]. AB - Correct diagnosis in patients with side effect during benzylpenicillin (penicillin G) treatment is highly problematic. It is accepted, that hypersensitivity to penicillin is overdiagnosed. Very little qualitative information is available on the diagnostic and differentiation of penicillin allergy. On present stage it is not techniques permitting to exclude anaphylaxis. This paper describe topical knowledge about risks administration of benzylpenicillin to patients suspected of penicillin hypersensitivity. Precious advice in diagnostic procedures and therapeutic management was given. PMID- 11503268 TI - [Nicotine and endocrine system in women]. AB - Cigarette smoking continues to be one of the world's most important substance abuse problems. The widespread and persistent use of cigarettes is caused by the addictiveness of nicotine. A variety of human and animal studies have shown that nicotine can induce changes in hormonal balance in women. A clear understanding of the pharmacokinetics and metabolism of nicotine may support changes in smoking behavior and exposure to tobacco toxins in women and may aid in the development of nicotine replacement products for smoking cessation. This review examines the present state of knowledge concerning the effects of nicotine on the female endocrine system. The first part of this article briefly summarizes recent findings from studies on nicotine metabolism. The second and third parts provide an overview of nicotine effects on hormone levels and the functional consequences of these effects in non-pregnant and pregnant women. In the final part, the influence of hormonal factors and pregnancy on metabolism and the pharmacological effects of nicotine are discussed. PMID- 11503269 TI - [Current opinions on immunological processes in rheumatoid arthritis during pregnancy]. AB - The essential in pathogenesis of RA is induction of incorrect immunological response against synovial and connective tissue antigens, which depends of CD4+ T cells activation by specific antigen. This stimulation leads to releasing Th1 lymphokines. The most important cytokine is TNF-alpha. An increased level of TNF alpha, IL-1, IL-6, GM-CSF, IL-8 was observed in patients with RA. PDGF, FGF, TGF, C-X-C a chemokines (IL-GRO-alpha, ENA78) and CCb chemokines (RANTES, MCP1 MIP1 alpha) are also involved in synovial hyperplasia in RA. During a pregnancy a clinical improvement in women with RA is frequent. The reason of this fact is probably connected with Th2 predominance (IL-4, IL-10) caused by presence of fetal tissues. Specific, cell-mediated immunity is suppressed and changed to Th2 by progesterone and PGE2. During a pregnancy a higher sensitivity of lymphocytes to progesterone was found. Progesterone stimulates T cells to PIBF production, which decreases NK activity. Th2 cytokines (Il-6, IL-10, IL-13, TGF) are expressed on decidua and inhibit secretion of Th1 cytokines (IL-2, INF gamma, TNF alpha, IL-1 alpha, IL-1 beta). Immunosuppression caused by pregnancy probably decreases inflammatory and destructive reactions in tissues women with RA. The first attack of this disease frequently observed during puerperium is connected with a high level of prolactin and a low of estrogens, which causes a increased release of IL-2 and has a main influence on initiation and increasing of inflammatory process in RA. PMID- 11503270 TI - [Clinical application of cyclooxygenase-2 inhibitors]. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) are a group of compounds with similar therapeutic and side effects. Their therapeutic effects depend on blockade of prostaglandin synthesis through enzyme cyclooxygenase (COX) inhibition. Two isoforms of the enzyme cyclooxygenase have been identified: COX-1 and COX-2. Selective COX-2 inhibitors i.e. meloxicam, nimesulid, etodolac or highly selective COX-2 inhibitors i.e. celecoxib, rofecoxib have antiinflammatory and analgesic properties with less or no gastrointestinal or other NSAIDs-typical adverse effects. Highly selective COX-2 inhibitors may also be active in colonic polyposis, colorectal cancer and Alzheimer's disease. PMID- 11503271 TI - [Vitamin C: antioxidant barrier]. AB - There is growing interest concerning the role of vitamin C in physiological conditions. In this paper L-ascorbic acid is discussed in detail including its chemical activity and biological properties. The primary goal of this review is to present the chemical characteristics of this substance and to discuss its relation to various biological functions of vitamin C, mainly as an antioxidant barrier. PMID- 11503272 TI - Potential application of immobilized and perfused hepatocytes in environmental toxicology studies. AB - Conventional cellular models have contributed significantly to the understanding of many aspects of cell physiology and molecular biology. In these models cells are metabolically less active, due to the inefficient oxygenation and waste product buildup. Therefore perfusion methods for the cells are expected to improve cell activities. Cells have to be fixed in or on an appropriate inert carrier or support, which enables cellular perfusion, maintains integrated cellular functions and makes a bioreactor. Since isolated hepatocytes are extensively used in biomedical studies including those dealing with environmental pollutants or toxins and in xenobiotic biotransformation investigations, an efficient hepatocyte perfusion model has to be available for researchers. This research article is focusing on the value of hepatocyte immobilization as a laboratory bioreactor model and is shedding light on its potentiality in research related to public health. We demonstrate the application of this cellular model as a means to study representative phase I and phase II biotransformation reactions using hexobarbital hydroxylation and 7-ethoxycoumarin deethylation and 4-chloro 2-dinitrobenzene glutathione. Both phase I and phase II drug biotransformation in hepatocytes was demonstrated in this study non-destructively to the cells and in an efficient way. In spite of the aforementioned advantages, immobilized hepatocytes yet have relatively limited applications compared to conventional hepatocyte cellular systems. Reasons for this discrepancy are discussed. This cellular system may become popular due to the better performance of immobilized hepatocytes as compared to conventional hepatocyte culture and due to economic and ethical reasons. Naturally its applicability will cover several biomedical areas including basic research in environmental toxicology and other public health issues. PMID- 11503273 TI - Frequency of occurrence of cytomegalovirus and Chlamydia pneumoniae in lymphocytes of atherosclerotic patients. AB - Forty-three blood samples from atherosclerotic donors and 28 samples from normal individuals were analyzed to determine the frequency of occurrence of cytomegalovirus (CMV) and Chlamydia pneumoniae DNA sequences in lymphocytes of Saudi Arabian donors using Polymerase Chain Reaction (PCR). In non atherosclerotic donors, no CMV DNA was detectable and only one sample was positive for C-pneumoniae DNA sequences. Of the 43 atherosclerotic patients, 22 were infected with CMV, 23 were infected with C-pneumoniae and 11 showed no infection. Thirteen of the 43 donors showed simultaneous infection with both CMV and C-pneumoniae. These results demonstrate that atherosclerotic patients are more frequently infected with CMV or C-pneumoniae or both. PMID- 11503274 TI - Injuries among children--some cost considerations in USA. AB - Unintentional and intentional injuries are a major cause of morbidity and mortality among children under the age of 19. The large number of premature death and disabilities due to injury and accompanying high economic cost, including public sector expenditures, emphasize the need to reduce the burden of injury. Productivity losses are greater from injury than from the three other leading causes of death--heart disease, stroke and cancer. Implementation of known injury control interventions can substantially reduce the incidence, severity, and accompanying cost of injury. Because financial resources are limited, decision makers need to consider the cost and effectiveness of alternative prevention strategies. Injury causes with the highest cost per injury could make a priority, especially within maternal and child health programs; as an alternative, projected total injury cost could determine program priority. PMID- 11503275 TI - Smoking and damages of reproduction: evidence of ELSPAC. AB - The international longitudinal prospective ELSPAC study also includes women from the Czech Republic: the results sum up data from 4530 women from City of Brno and District of Znojmo who had different exposure to active and passive smoking. Anamnestic data were collected by fulfilling the internationally unified questionnaires during pregnancy and at the 2nd month after delivery. Gynaecologists, obstetricians and pediatricians, using the unified procedures, were collecting objective data in the course of the pregnancy, delivery, monitoring health status of women and newborns. Significant differences were found between smoking and non-smoking women in the prevalence of alcohol and marihuana consumers both before and during pregnancy and after delivery. In personal health history, smoking women more often reported their self-perceived poor health, panic and loss of self-control during the delivery. In the current pregnancies, smoking women suffer more often with placenta praevia, placental abnormalities, fetal growth retardation and fetal malformations. On the other hand, the prevalence of preeclampsia was decreased among smokers. The children of mothers who were moderate/heavy smokers during pregnancy, had on average by 245 g lower birth weight, 1.22 cm shorter body length and by 0.66 cm smaller head circumference than those of non-smokers. These measures were on average also decreased in the groups of newborns whose mothers were light smokers or non smokers heavy exposed to environmental tobacco smoke. On the contrary, the average gestational age was similar both in case of active smoking, passive smoking, and non-smoking women. In spite of the fact that most of smokers would give up smoking after getting pregnant, it is necessary to place the antismoking interventions and nutrition advisory service within the routine duty of physicians. PMID- 11503276 TI - The outcome of infertility treatment by in-vitro fertilisation in smoking and non smoking women. AB - The former and current smoking habits of 159 patients treated for infertility by means of IVF/ET technique were observed. The data were verified by a cotinine examination in urine. The basal levels of gonadotropines before treatment, the process and outcome of ovarian stimulation, oocyte fertilisation and incidence of pregnancy were observed. There were no differences between smokers and non smokers in terms of their age and professional risks; smoking correlated with a lower level of education. The overall response of active smokers to hormonal stimulation was worse than that of non-smokers: a lower number of mature follicles (12.3 vs. 16.2) and a lower number of oocytes were gained (7.3 vs. 10.9). There was a correlation between the age of the smokers and the higher consumption of FSH needed for stimulation and lower levels of 17 beta-estradiole. The number of fertilised oocytes in smokers was significantly lower (p < 0.01; 68.2% vs. 47.8%). A negative correlation between the duration of exposure to cigarette smoke and fertilisation rate was shown (p < 0.05). There were fewer embryos in smokers as compared with non-smokers (3.3 vs. 4.7). A total of 35 women became pregnant (i.e. 22.1%), of which 28.8% were non-smokers, 12.5% were occasional smokers, and none were regular smokers. The OR value in non-smokers was 1.48, and in occasional smokers 0.57. In this study, the negative influence of smoking on the outcome of treatment by IVF/ET technique was proved. The phasing out of smoking should be an integral part of human infertility treatment. PMID- 11503277 TI - Premature mortality in lung cancer as an indicator of effectiveness of tobacco use prevention in a gender perspective--a comparison between Poland and Sweden. AB - This contribution presents a comparative analysis of the probability of premature overall deaths and lung cancer mortality for men and women between one Nordic country--Sweden--and one country in transition in central Europe, namely Poland. Furthermore the study compares the pattern of smoking prevalence for both sexes in the two countries. Male lung cancer mortality is constant in Sweden during the last two decades, which is completely contrary to the trend in Poland with a long period of increase followed by a slow decrease during the last years. Lung cancer mortality for women in both countries is on a continuous increase and the rate is exactly the same. The premature mortality in lung cancer in Sweden in the age group of 20-44 years now is higher for women than for men. Swedish men have reduced their smoking habit strongly and far more than Polish men (17 respectively 42% 1999). The male use of moist snuff in Sweden is taken into consideration when discussing the low rate of lung cancer in Sweden. Concerning the pattern of tobacco use it is obvious that Swedish and Polish women during the last decades have had the same and slowly decreasing smoking prevalence (21 respectively 23% 1999). The conclusion is that the best way to control the lung cancer epidemic is to reduce smoking prevalence in the population. Lung cancer mortality in younger and middle aged groups is a good indicator of the success in tobacco control in a country as well for women as for men, especially in the middle age groups. PMID- 11503278 TI - Role of genetic factors in development and progression of non-Hodgkin's lymphomas. AB - Non-Hodgkin's lymphomas (NHL) are a heterogeneous group of neoplasms with wide variation in histologic features, immunologic phenotype, molecular abnormalities, clinical presentation, and disease progression. New molecular techniques have significantly increased understanding about the molecular background for development of NHL and it has been possible to identify typical genetic abnormalities in specific NHL subtypes. Some of the genetic changes and alterations in predisposing genes related to NHL are reviewed in this article. The reviewed information suggests that it is of great importance to look for reliable diagnostic tools for screening of population groups to identify individuals at high risk of lymphomas. The follow up of interaction of various predisposing genes and environmental factors in lymphoma pathogenesis is of considerable public health importance as well. In a pilot study, polymerase chain reaction-restriction fragment length polymorphism based genotyping assays were used to determine the frequency of polymorphisms in genes coding for various biotransformation enzymes in a case-control study comprised of 147 patients with NHL and group of age- and sex-matched unrelated healthy Czech individuals. Preliminary statistical analyses showed no association of CYP1A1-m1/m2, CYP2E1 c1/c2, GSTM1null, and GSTT1null genotypes with incidence and status of NHL. On the other hand significant differences in distribution of genotypes of CYP2E1 intron 6, EPHX-exon 3, and GSTP1-exon 5 were observed between cases and controls. Thus it seems that study of co-segregation of particular genotypes of biotransformation enzymes with higher risk of lymphoproliferative disorders may prove to be very useful approach in elucidation of the etiology of malignancies of lymphoid origin. PMID- 11503279 TI - Epidemiology of invasive meningococcal infections of children in Slovenia during 1993-1999. AB - Invasive strains of Neisseria meningitidis isolated from sterile body fluids of children aged 0-14 years were intentionally collected in nine Slovenian laboratories, and sent to the laboratory of the National Institute of Public Health, Ljubljana for final evaluation. From 1993-1999 we collected 53 invasive strains from children presented in this study. The incidence rate of invasive diseases in children shows that there is a rapid fall from 4.74/100,000 in 1993 to 0.90/100,000 in 1999. Invasive diseases predominated in children in the age group 0-1 year (19.6 cases per 100,000 children); followed by the age groups 2-4 years, 5-9 years and 10-14 years. The results of serogrouping indicate that the largest proportion of meningococci isolated (90%) were group B, while 8% of isolates were group C and only 2% group Y. The serotype/serosubtype distribution shows that the most frequent serotype (excluding 19 non-typable NT strains) was serotype 22, which was expressed in 9 strains and the most frequent serosubtype (excluding 11 NST strains) was P1.5,2 found in 7 strains. The most frequently isolated strains (excluding NT/NST strains) were B:22:P1.14; B:NT:P1.5 and B:NT:P1.16. Thirty-one strains collected during 1993-1995 were analysed by multilocus enzyme electrophoresis (MEE). There were 25 electrophoretic types (ETs) among 31 strains; the strains were very heterogeneous with only four ETs being represented by more than one strain. Study shows that meningococcal disease is still endemic in Slovenia. PMID- 11503280 TI - Does alcohol protect against ischaemic heart disease in Bulgaria? A case-control study of non-fatal myocardial infarction in Sofia. AB - BACKGROUND: Many observational studies have shown that alcohol consumption is associated with a reduced risk of ischaemic heart disease (IHD). IHD mortality has generally fallen in established market economies but not in some countries of Eastern Europe. Since the level of consumption of saturated fat does not explain these differences in trends, other associations with risk need to be explored. We investigated whether alcohol consumption also presents a U or J-shaped association with IHD risk in a case-control study in Bulgaria. METHODS: Cases (n = 155) were admissions to the cardiology unit, Central Clinical Hospital, Sofia, aged 45 to 69, with confirmed diagnoses of ischaemic heart disease. Controls (n = 154) were concurrent admissions for minor elective surgery. Measurements were made of blood pressure, height and weight and a blood sample was taken around three days after admission. Subjects were interviewed before discharge and asked about the type and amount of alcohol they consumed. RESULTS: Reported alcohol intake demonstrated a J-shaped association with the risk of IHD. The odds ratio (adjusted only for age and sex) was 0.67 (95% CI 0.34-1.28) for patients reporting 0.01-18 g/d of alcohol consumption daily, and 0.36 (95% CI 0.18-0.73) for 18.01-36 g/d, compared to patients reporting to be abstainers. The associations with alcohol intake remained statistically significant and unaltered after adjustment for established IHD risk factors: HDL cholesterol, body mass index, systolic blood pressure, family history, education, physical activity and risk factors significantly related with IHD: fruit and vegetables consumption, type of fat used in cooking, bread consumption. CONCLUSIONS: Our results are consistent with previous studies showing a J-shaped association between alcohol intake and IHD risk. The highest protective effect we observed for levels of alcohol intake 18.01-36 g/d, which corresponds to 100-200 ml wine or 1-2 beers, or little more than 50-100 ml spirits. PMID- 11503281 TI - Pilot study of risk behaviour, voluntary HIV counselling and HIV antibody testing from saliva among inmates of prisons in Slovakia. AB - OBJECTIVE: To implement a pilot study of risk behaviour and HIV infection using HIV antibody testing from saliva to improve the situation as regards HIV/AIDS infection in prison institutions in the Slovak Republic. MATERIAL AND METHODS: The study comprised adult and juvenile males of grade one correction categories and prisoners from the prison for juveniles in Martin, as well as females prisoners in Nitra. Preventive activities were implemented in May 1998 in the form of discussions concerning topics related to HIV/AIDS infection. Saliva was collected for the presence of HIV antibodies and a questionnaire regarding sexual practice was completed. RESULTS: 32 persons [8 adult males (25%), 6 juvenile males (18.7%) and 18 females (56%)] were voluntarily tested for the presence of HIV antibodies in saliva. Nobody was HIV-positive. 75 persons (20 adult males, 30 juvenile males and 25 females) were involved in the study of risk behaviour. 40.8% participants had primary education, 28.2% secondary education, 2.8% were students of universities and 28.2% were apprenticies. 60% inmates (mostly females) were religious. Juvenile males reported the highest number of partners while females the smallest (p < 0.001). The more partners were reported by respondents, the lower was condom usage (p < 0.07). 47.6% females relied on credibility of partners, while 75% adult males and 50% adolescent males did not use protection. 0% females, 5% adult males and 10.3% juvenile males reported to have homosexual contacts outside prison while 19%, 5.6% and 8.3% in the prison, respectively. Paid sexual services were offered by 9.1% females, 15.8% adult males and 25% juvenile males. Outside prison adult and juvenile males used non sterile used syringes as well as tattooing more often than females (p < 0.07 and p < 0.04, respectively). CONCLUSION: The present study provides information on the results of HIV-antibody testing in saliva. The results are based on a study of risk behaviour and difficulties linked with HIV/AIDS prevention among prisoners. PMID- 11503282 TI - On the need to teach evidence-based preventive medicine to health professionals. AB - In the last decade evidence-based medicine (EBM) has become popular between clinicians and epidemiologists as a tool to facilitate the translation of scientific research into clinical practice. In public health, this approach is important for health protection, different levels of disease prevention, and health promotion practice. EBM concept applicable to public health follows the same principles, but some additional aspects have to be considered. For example, in preventive medicine, there is the strong need to compare the relative effectiveness of various interventions in prolonging survival or preventing the occurrence or complications of a disease, as it is crucial for setting preventive priorities. Usually, systematic reviews and meta-analyses quantify the effects of treatments on selected endpoints (health outcomes). However, in setting preventive priorities the reverse perspective is important. The judgement about preventive action cannot be done without an adequate consideration of ethical and social context. In view of emerging evidence-based medicine to health professionals became of paramount importance. In the paper, this course of action has been discussed as an aftermath of the EB training workshop organized in Pavia (Italy) by the International Center for Studies and Research in Biomedicine. PMID- 11503284 TI - Cholera: a re-emerging infection. PMID- 11503283 TI - Pattern recognition of respirable dust particles by a back-propagation artificial neural network. AB - A back-propagation neural network was used as a pattern recognition tool for LAMMA mass spectral data. Standard EPA source profiles were used as training and test data of the net. The elemental patterns (10 elements) of the sum of 100 mass spectra of fine dust particles were presented to the trained nets and satisfactory recognition (> 50%) was obtained. PMID- 11503285 TI - Awake respiratory function in patients with the obstructive sleep apnoea syndrome. AB - INTRODUCTION: The flow-volume curves of patients with obstructive sleep apnoea (OSA) obtained during the awake state are frequently abnormal. OBJECTIVE: To determine 1) the relationship between the awake respiratory function and the severity of sleep-disordered breathing in a group of Malaysian patients with the OSA syndrome and 2) the frequency of flow-volume curve abnormality in these patients. MATERIALS AND METHODS: A retrospective analysis of the data from respiratory function tests during wakefulness and nocturnal polysomnography was performed on 48 patients with OSA. The severity of OSA was defined by the apnoea hypopnoea index (AHI) and the lowest oxygen saturation during sleep (SpO2nadir). RESULTS: AHI had a significant relationship with alveolar-arterial oxygen gradient (r = 0.34, p = 0.046) and SpO2nadir (r = -0.49, p < 0.001) but not with any anthropometric parameter or the other awake respiratory function variables measured. SpO2nadir had a significant relationship with body mass index (r = 0.54, p < 0.001), neck circumference (r = -0.39, p = 0.013), awake room air PaO2 (r = 0.61, p < 0.001), alveolar-arterial oxygen gradient (r = -0.41, p = 0.015) and baseline supine SpO2 (r = 0.53, p < 0.001). There was no correlation between SpO2nadir and any spirometric or static lung volume parameters. The maximum inspiratory and maximum expiratory flow-volume curves of 26 patients (54%) showed a ratio of forced expiratory flow to forced inspiratory flow at mid-vital capacity (FEF50/FIF50) greater than one. In addition, flow oscillations (the "sawtooth" sign) were noted in the inspiratory and/or expiratory flow-volume curves of 21 patients (44%), 9 of whom did not have an FEF50/FIF50 > 1. Altogether, the maximum flow-volume curves during wakefulness of 35 (73%) of the 48 patients showed variable upper airway obstruction and/or flow oscillations. However, the presence of these two upper airway abnormalities, either occurring alone or together did not have an effect on the severity of OSA as measured by the AHI or SpO2nadir. CONCLUSIONS: Abnormalities of the flow-volume loop consistent with inspiratory flow limitation and/or upper airway instability during wakefulness are common in patients with the OSA syndrome. The degree of oxygen desaturation during sleep in these patients is related to their awake oxygenation status. PMID- 11503286 TI - Appendicular metastasis from carcinoma cervix. AB - A young lady who was treated for early squamous cell carcinoma of cervix presented with perforated appendicitis. Appendicectomy was done and the histopathology was reported as metastatic squamous cell carcinoma. Squamous cell carcinoma of the cervix metastasizing to the appendix is extremely rare and we previously unreported. PMID- 11503287 TI - 'Thermal reflex therapy' a danger for diabetic feet. PMID- 11503289 TI - A view for patients. PMID- 11503288 TI - Diabetic retinopathy in the community. PMID- 11503291 TI - Factors influencing the outcome of arthrodesis for congenital kyphosis and kyphoscoliosis. AB - Sixty-five patients with congenital kyphosis and kyphoscoliosis who underwent spinal arthrodesis are reviewed to determine the factors that influenced the outcome of arthrodesis. Mean follow up after surgery was 6 years and 6 months with all patients having a minimum of 2 years follow up. A satisfactory outcome, or a stable arthrodesis was defined as a loss of correction of less than 10 degrees from the time of surgery till review. An unsatisfactory outcome, or unstable arthrodesis was considered when there was more than 10 degree loss. Type of vertebral anomaly and type of arthrodesis procedure were significantly influenced stability of arthrodesis, whereas age when arthrodesis was performed and size of curve at surgery were not significantly related to stability of arthrodesis. PMID- 11503290 TI - Risk of tuberculosis to healthcare workers. PMID- 11503292 TI - Notification of occupational and work-related diseases and poisonings in Malaysia, 1997-1998. AB - In 1997, the Ministry of Health Malaysia introduced a surveillance programme for occupational and work-related diseases including poisonings for cases seen in government health facilities. Between June 1997 and November 1998, there were 36 cases of respiratory disease and 95 cases of poisoning by chemicals and pesticides notified while skin diseases were 108 cases. Respiratory diseases reported were predominantly occupational asthma (25%), pneumoconiosis (17%) and infections (39%). The commonest reported skin disease was contact dermatitis (87%). The commonest causes of occupational poisonings were paraquat (19%), organo-phosphates (16%), agro-chemicals excluding pesticides (15%) and gases (10%). The number of cases reported is still relatively few compared to data from other countries, suggesting that there is still considerable under reporting. PMID- 11503293 TI - The use of an in-house biotin-avidin linked immunosorbent assay to detect Aspergillus antigens in sera of immunocompromised patients. AB - A biotin-avidin-linked immunosorbent assay was developed to detect Aspergillus antigens in sera of immunocompromised patients. The assay was based on a double antibody sandwich ELISA using polyclonal antibodies raised against water-soluble antigens of Aspergillus fumigatus. Aspergillus antigens were positive in sera of 9 of 16 (56%) patients who were studied prospectively and in 13 of 73 (19%) patients studied retrospectively. The 9 prospectively studied patients who were antigen positive were febrile neutropenic hematological malignancy patients who exhibited a high risk of acquiring invasive aspergillosis. PMID- 11503294 TI - A retrospective study of death among patients treated for tuberculosis in the Klang Chest Clinic for the year 1999. AB - A retrospective epidemiological study was done on 41 deaths among patients treated for TB in the Klang Chest Clinic for the year 1999. The findings revealed a male preponderance of TB deaths with Indians having the highest case fatality rate (8.6%). The majority of deaths occurred within the 25-44 year age group. Of those cases diagnosed as Pulmonary TB, 69% were diagnosed as far-advanced, at presentation, based on chest x-rays reported. Eighty three percent of these patients were still on treatment when they died. HIV was the most common co existing disease condition and was implicated in 14.6% of the deaths. The recommendations include implementation of clinical guidelines on TB screening for HIV patients, a study to evaluate the effectiveness of the current TB surveillance programme and for chest x-rays of all sputum smear negative patients to be reported by the radiologist to reduce the risk of misdiagnosis. PMID- 11503295 TI - Detection of Vibrio cholerae 01 from aquatic environment in Sarawak. AB - The detection of Vibrio cholerae 01 from the aquatic environment of Daro and Bintulu in Sarawak was carried out following an outbreak of cholera. Conventional culture methods and detection of ctx gene by polymerase chain reaction technique were carried out on 80 water samples. Only one sample was positive by culture methods while 8 were positive by PCR. DNA finger printing by pulsed-field gel electrophoresis showed that the clinical isolates in Daro and Bintulu were genetically identical while the environmental isolate was closely related. Recovery of Vibrio cholerae by culture method is poor and newer methods of detection should be developed. PMID- 11503296 TI - Bone transport in the management of fractures of the tibia. AB - A review was carried out in 21 cases of bone transport in the tibia done between May 1995 and December 1997. These were done for the treatment of compound (Grade IIIB) fractures with extensive bone and soft tissue loss and in infective non unions of the tibia using the Ilizarov technique and ring fixator. In 5 cases, 2 or 3 additional procedures were needed such as tendo achilles (TA) lengthening, bone grafting, revision of construct or revision of scar at the docking site. Average resection of infected bone was 5.2 cms in the infective non-union group and average bone and soft tissue loss was 8 cms in the compound Grade IIIB fracture group. The defect was bridged and regenerate bone occurred in all the cases except one. Union was achieved in all the cases although 10 needed bone grafting. Infection was eradicated in all the cases. Limb length discrepancy was corrected in all the cases except three. Mean duration of treatment was 12 months. PMID- 11503297 TI - Trichuris Dysentery Syndrome: evidence that it may be underdiagnosed in Kelantan. AB - The prevalence rate of trichuriasis in children in certain areas of Kelantan is high. However the Trichuris Dysentery Syndrome (TDS), a condition in children characterized by chronic diarrhoea, stunting and anaemia is said to be rare. A recent change in policy at our institution to lower the threshold for undertaking colonoscopy in children with chronic diarrhoea resulted in the detection of 6 cases of TDS in less than a year. The median age was 7 years (range 3-13) and the median duration of symptoms before diagnosis was 2 years (range 1-8). The insidious nature of TDS and the low level of awareness of this condition even among health care workers may result in considerable underdiagnosis. PMID- 11503298 TI - Ambulatory surgery and anaesthesia in HUKM, a teaching hospital in Malaysia: the first two years experience. AB - Ambulatory or day care surgery is still in its infancy in this part of the world. Our newly built university affiliated hospital started its Day Surgery Centre in February 1998. It is the first multidisciplinary ambulatory surgery centre in a teaching hospital in the country. It caters for Orthopaedic surgery, Urology, Plastic surgery, Otorhinolaryngology, General surgery, Paediatric surgery and Ophthalmology. We have done 2,604 cases and our unanticipated admission rate is less than 2%. There has been no major morbidity or mortality. The problems of setting up a multidisciplinary ambulatory centre in a teaching hospital are discussed. PMID- 11503299 TI - Breastmilk feeding status and weight gain of low birth weight infants in a Neonatal Intensive Care Unit. AB - A prospective observational study of feeding in low birth weight (LBW) infants with birth weight (BW) of at least 1.8 kg admitted to the Neonatal Intensive Care Unit (NICU) showed that nearly 80% of mothers provided expressed breastmilk (EBM) and a further 14% breastfed their infants before discharge. Weight gain was overall poor at a mean of 9.48 +/- 7.82 grams per kg per day with those on predominant EBM feeding (EBM > 70%) doing worse than those on predominant preterm formula (PTF) feeding (EBM < 31%), weight gain being 5.40 +/- 6.88 and 11.10 +/- 8.15 grams per kg per day respectively (p < 0.01). Weight gain was also poorer (7.72 +/- 5.55 grams per kg per day) in patients with respiratory distress syndrome (RDS) compared to those who did not have RDS (12.02 +/- 9.58 grams per kg per day). p < 0.05. Incidence of infants < 10th centile body weight at birth was 16.8% and at discharge was 69.1%. PMID- 11503300 TI - Feeding practices of infants delivered in a district hospital during the implementation of Baby Friendly Hospital Initiative. AB - A study on infant feeding practices was conducted during the implementation of the Baby Friendly Hospital Initiative (BFHI) in a district hospital. The aim was to identify which population subgroups had lower breastfeeding rates at 4 months and the effect of attendance of antenatal breastfeeding education on breastfeeding practices. All infants delivered in May 1996 were followed-up. 204 respondents were analyzed. This study demonstrated a higher exclusive and any breastfeeding rates at 4 months than some other studies. (48% and 76% respectively). It was found that the Malays were more likely to be breastfeeding exclusively at 4 months (72%) than the Indians (32%) and the Chinese (4%). (P < 0.01). There were more non-working mothers breastfeeding exclusively at 4 months than working mothers. (60% versus 26%) P < 0.01. Antenatal breastfeeding education in the form that was given appeared to improve breastfeeding rates at 4 months. Future efforts to promote breastfeeding should target the Chinese mothers and the working mothers. PMID- 11503301 TI - Crash simulation of lower limb with motorcycle basket. AB - Lower limb injuries are the main cause of temporary and permanent disability among motorcyclists in Malaysia. They cause non-fatal but serious injuries requiring hospitalisation. Detailed studies on factors influencing lower limb injuries are justified in an attempt to reduce the occurrence of these injuries. This study presents a computer simulation of the crash behaviour of the basket of a small-engined motorcycle with the lower limb using finite element (FE) methods. The results suggest that the extensive deformation of the motorcycle basket may reduce the risk of injury to the lower limb. The behaviour of the basket during collision is analogous to the crumple zone of automobiles. PMID- 11503302 TI - Outcome of 235 renal transplant recipients followed up at ministry of health hospitals in the State of Johor. AB - Retrospective analysis was done on 235 recipients, 133 males and 102 females, who were transplanted between 25th September 1979 and 25th June 1999. 85.1% were Chinese, 7.7% were Indians and 7.2% Malays. 23% (54) were living related renal transplants (LRRT) all except 5 done at Hospital Kuala Lumpur. 60% (141) were living unrelated donor renal transplants (LURT) done in India. 17% (40) were cadaveric transplants (CADT) (all done in China except 2 at Hospital Kuala Lumpur and one in London). 97% (228) were first transplants. Primary renal disease was unknown in 69.4%, 17% (40) glomerulonephritis, 5.5% diabetic nephropathy and 8.1% 19 others. All were on prednisolone, 93.2% were on azathioprine and 96.6% were on cyclosporin A. The acute rejection rate was 23.4% (55 episodes). Patient survival was 88% at five years and patients alive with functioning graft was 84% at 5 years. LRRT had significantly better survival compared to LURT. 34 grafts were lost to chronic allograft nephropathy. 46 recipients died (33 died with functioning graft). PMID- 11503303 TI - Ectrodactary, ectodermal dysplasia, and cleft lip-palate syndrome. AB - The EEC syndrome (ectrodactyly, ectodermal dysplasia and cleft lip-palate) is a rare disorder inherited as an autosomal dominant trait or can occur sporadically. We describe a case of this syndrome with chronic ocular surface disorder secondary to abnormalities of ocular adnexia. PMID- 11503304 TI - Phenothiazine deposits in corneo and lens. AB - Phenothiazines have been associated with various ocular adverse effects, in particular cataract and corneal deposits. Several effects are vision threatening. Awareness of these effects, in particular by Psychiatrists and Ophthalmologist, is essential for prompt diagnosis and appropriate treatment. We report two patients presented to us with corneal deposits and lenticular opacities following long term use of Phenothiazines. PMID- 11503305 TI - Pancreatic tuberculosis presenting with recurrent acute pancreatitis. AB - Tuberculosis, in its extrapulmonary form, though emerging as a common clinical problem, rarely affects the pancreas. Its indolent course, vague symptomatology along with its non-specific laboratory and radiographic findings call for greater vigilance. We report a case of pancreatic tuberculosis, previously managed as recurrent alcohol related pancreatitis which showed symptomatic improvement following commencement of antituberculosis drugs. The diagnosis of pancreatic tuberculosis in this case was based on the abdominal CT scan findings, response to anti-tubeculous chemotherapy and overall laboratory and radiological work-up. PMID- 11503306 TI - Knee crepitus from an unusual source. AB - A case report of a missed appendicitis presenting with abdominal wall necrotising fasciitis which extended up to the right knee. This subcutaneous collection in the prepatella region of the right knee presented as a crepitus and mimicked an intraarticular pathology. PMID- 11503307 TI - [Helping the patient to decide]. AB - Since 1998, a second opinion practice in Amsterdam has tried to find a method for providing the patient with decision support. Current literature is studied and discussed with the patient and support is given with the interpretation of medical information. This also means providing insight into the peculiarities of doctors' diagnostic and therapeutic reasoning. Informed consent or the decision to refuse treatment should be based on sound arguments and balanced information. Due to a disc hernia, a 60-year-old man suffered from immobilizing pain and a loss of strength in his right leg. Surgery was proposed but the patient had found additional information on the Internet, which the neurosurgeon was not prepared to discuss. A 40-year-old woman suffering from incontinence was offered periurethral collagen injection therapy. She had found a medical article on this subject for which she sought an explanation. On the basis of this discussion questions were formulated which she could pose to her urologist. A 70-year-old woman with shoulder pain, who was not satisfied with the pain medication prescribed by her general practitioner, came to discuss the possibilities of second opinion and referral to a specialist. A 29-year-old woman had consented to a surgical correction of a urethral stenosis, now wondered whether she could be treated laparoscopically. She phoned for advice and to find out what sort of questions she should pose to her urologist. PMID- 11503308 TI - [Decision support in primary care--a vertical second opinion]. AB - This comment concerns a specific second opinion practice in Amsterdam. In this practice a physician limits her consultations by optimally preparing patients for their subsequent encounter with the responsible clinician. It is argued that this type of practice must be discouraged, as due to its limited goal, lack of personal responsibility and subordination of 'confidence' in doctor-patient relationships, it constitutes a strongly deviant form of clinical practice. PMID- 11503309 TI - [Medical research and the pharmaceutical industry. Uneasy bedfellows or a prenuptial agreement?]. AB - Recent publications in medical journals and the media seem to indicate that there are a number of major problems in the relationship between researchers and the pharmaceutical sector. Results of research allegedly are influenced or manipulated, the right to publish the results are taken away from the researchers, and all this for money. The underlying data do not seem to indicate that this is a very common problem, but rather a generalisation of incidents. To efficiently develop medicines in future requires an ever-growing amount of academic know-how that these companies simply lack; in modern pharmaceutical development there are many opportunities to connect academic and industrial research programmes, resulting in synergy rather than an academic sell-out. All parties concerned should make transparent and clear agreements. PMID- 11503310 TI - [Childhood cancers in the Netherlands (1989-1997)]. AB - In a recent report by the Society of Integral Cancer Centres in the Netherlands, attention was devoted to the incidence of cancer in children and the mortality arising from this. In recent years the growing diagnostic and therapeutic possibilities have changed the perspective of childhood cancer enormously. Based on a careful classification and clinical staging, national and international investigations have resulted in new and successful therapeutical strategies. Overall prognoses of childhood cancer have improved dramatically from a 5-year survival rate in the 1960s and 1970s of less than 30%, to an 8-year survival rate of more than 70% between 1989 and 1997. However, this success means that more investigations into the long-term effects of childhood cancer and its treatment are needed. In a study at the Amsterdam University Hospital, 700 adult survivors of childhood cancer were reinvestigated of whom over 75% experienced one or more clinically relevant long-term effects. A continuous survey for long-term effects is needed for the development of new therapeutic strategies, which allow children treated for cancer to develop with the same possibilities in life as their healthy peers. PMID- 11503311 TI - ['Variant of Creutzfeldt-Jacob disease and blood transfusion'; report of the Dutch Health Council]. AB - The new variant form of Creutzfeldt-Jakob Disease (vCJD), which has been diagnosed in about 100 patients--mostly in the United Kingdom (UK)--is considered to be associated with the consumption of beef contaminated with the agent bovine spongi-form encephalopathy (BSE). Although no cases of vCJD have been reported until now in the Netherlands, large quantities of beef have been imported from the UK in previous years; furthermore about 17 cattle with BSE have been detected in the Netherlands. Concern about the possible transmission of vCJD via blood and blood-products has led to a number of countries taking precautionary measures. Following questions raised by the Minister of Health, Welfare and Sport, the Health Council of the Netherlands issued a report to address the need for certain precautionary measures such as the leukodepletion of blood and the exclusion of donors at risk for vCJD. The Health Council recommends the routine leukodepletion of cellular blood components. The exclusion of donors who have resided in the UK for six or more months during the period 1980-1996, was considered to be insufficient to contribute to risk reduction. The Minister has recently decided to follow these two recommendations. However, she is of the opinion that the Health Council's recommendation to exclude all donors who have previously been transfused with cellular blood components is unnecessary. A common European position regarding such precautionary measures is deemed to be necessary. This would allow the exchange of blood components between countries and would also prevent donors, patients and the public at large from being confused or uncertain about the safety of blood components. PMID- 11503312 TI - [Epidemiological developments and insights in chronic inflammatory bowel diseases]. AB - In the Western world, a sharp increase in the incidence of both Crohn's disease and ulcerative colitis was observed during the sixties and seventies, after which it reached a plateau at around 7 and 12 new cases per 100,000 inhabitants per year, respectively. Both chronic inflammatory bowel diseases usually commence between 15 and 30 years of age. Most patients have a normal life expectancy. A multifactorial model has been proposed to explain the pathogenesis, in which there seems to be an interaction between genetic predisposition, environmental factors and an altered immune response (occurring either after or because of the disease). Smoking is a clear risk factor in the development of Crohn's disease and it also negatively influences the clinical outcome. On the other hand, stopping smoking is a risk factor in the development of ulcerative colitis. PMID- 11503313 TI - [Limited role of naltrexone in the treatment of opiate addiction]. AB - Naltrexone has been used for many years as an opiate antagonist for maintenance treatment to prevent relapse in opiate addiction and, more recently, for rapid opiate detoxification with and without general anaesthesia. Naltrexone is useful for rapid detoxification without anaesthesia. However, the detoxification procedure under anaesthesia lacks a scientific basis. There is no clear evidence of the efficacy of naltrexone maintenance treatment. Poor compliance and possible receptor sensitisation means there may be a potentially increased risk of mortality through opiate overdose following cessation of naltrexone treatment and relapse into addiction. PMID- 11503314 TI - [From gene to disease; from SHOX to Leri-Weill dyschondrosteosis, Turner syndrome and idiopathic short stature]. AB - Leri-Weill dyschondrosteosis is a pseudodominant hereditary skeletal dysplasia associated with haploinsufficiency of the SHOX gene. The SHOX gene is located on the pseudoautosomal region of both the X-chromosome and the Y-chromosome, and belongs to the homeobox genes, which code for transcription factors involved in early foetal development. Mutations in the SHOX gene, especially deletions, are detected in approximately 60% of patients with Leri-Weill dyschondrosteosis. The SHOX gene is also involved in skeletal abnormalities in Turner syndrome and possibly plays a role in idiopathic short stature. PMID- 11503315 TI - [Diagnostic image (48). Linitis plastica]. AB - In a 67-year-old woman with recurrent vomiting the gastric folds did not disappear after air insufflation. Linitis plastica was diagnosed and stomach resection was carried out. PMID- 11503316 TI - [Local tumor control and (disease-free) survival after surgery with pre- and intraoperative radiotherapy for primary non-resectable rectal carcinoma and local recurrence]. AB - OBJECTIVE: Survey of the results of multimodality treatment for primary irresectable rectum carcinoma and local recurrence of rectal cancer. DESIGN: Retrospective. METHODS: During the period 1 February 1994 to 31 August 1999, 43 patients with locally advanced primary rectal cancer (25 men and 18 women; mean age: 64 years (range: 36-86)) and 53 patients with a local recurrence (33 men and 20 women; mean age: 61 years (39-82)) were treated with a multimodality treatment: i.e. preoperative radiotherapy (doses 50.4 Gy, or 30.0 Gy in the case of reirradiation), extensive surgery and intraoperative radiotherapy (doses 10 17.5 Gy). This treatment took place at two hospitals in the Netherlands, the Catharina Hospital in Eindhoven and, since 1997, the Daniel den Hoed Cancer Centre in Rotterdam. In 2000 data were collected for the local control and (disease-free) survival; these were analysed using the Kaplan-Meier method. Patients also completed a questionnaire about the quality of life at a median period of 14 months (range: 4-60) after the operation; the response level was 96% (76/79). RESULTS: After 3 years, the local control, disease-free survival and survival rates for the locally advanced primary rectal cancer group were 74%, 60% and 55% respectively, and for the locally recurrent rectal cancer group 64%, 34% and 50% respectively. Tumour resection with microscopically negative margins had a statistically significant positive effect on the local control and disease-free survival in both groups as well as on the survival in the locally advanced primary patient group. Seven of the 96 patients (7%) died as a result of complications. Of the patients with a primary irresectable carcinoma or a local recurrent tumour who completed the questionnaire the results were as follows: 56% and 63% respectively had been able to resume employment, 53% and 59% respectively had been able to resume their previous lifestyle, 15% and 27% respectively indicated radicular pain as a new symptom, 26% and 46% respectively stated problems with walking, 42% and 44% respectively stated problems with urinating and 59% and 52% respectively a reduction in sexual activity. PMID- 11503317 TI - [Clinical algorithm for cerebrospinal fluid test of 14-3-3 protein in diagnosis of Creutzfeldt-Jacob disease]. AB - OBJECTIVE: To study whether an algorithm that includes additional diagnostic information could increase the specificity of the 14-3-3 protein testing in patients suspected to suffer from Creutzfeldt-Jakob disease (CJD). DESIGN: The development of a diagnostic algorithm. METHOD: The 14-3-3 protein was tested in the cerebrospinal fluid from 69 consecutive patients suspected of having CJD. On the basis of a former study and literature research, a diagnostic algorithm was constructed, which restricted the indication for performing the 14-3-3 protein test. RESULTS: By taking into consideration the findings of neuroimaging and routine cerebrospinal fluid examination prior to 14-3-3 testing, the specificity increased to 97% (95%-CI: 85.5-99.9) thus changing the prior probability of having CJD of 35% to a posterior probability of 75-100%, in the case of a positive test result. CONCLUSION: Determining the presence of 14-3-3 protein is a highly sensitive and specific marker for sporadic CJD when used in combination with imaging and cerebrospinal fluid examination. PMID- 11503318 TI - [Six-month old girl with an anaphylactic reaction to rice flour, a rare food allergy]. AB - A 6-month-old girl had been admitted three times due to the sudden onset of respiratory and gastrointestinal symptoms, paleness and a reduced level of consciousness. Various laboratory tests (including allergy tests) as well as microbiological and radiological investigations did not establish the cause. Only when it was noticed that the incidents followed a change of diet, did it became obvious that a reaction to food components was the underlying cause. A provocation test revealed an anaphylactic reaction to rice flour. Food allergy is a widespread problem within paediatrics; rice flour appears to be hypoallergenic, but can lead to immunological responses. PMID- 11503319 TI - [Facial paresis in children; consider Lyme disease]. PMID- 11503320 TI - [Drug utilization by the elderly with depressive symptoms]. PMID- 11503321 TI - [Diagnostic image (32)]. PMID- 11503322 TI - [Safety of specific cyclo-oxygenase 2 inhibitors]. PMID- 11503323 TI - [Safety of specific cyclo-oxygenase 2 inhibitors]. PMID- 11503324 TI - [Safety of specific cyclo-oxygenase 2 inhibitors]. PMID- 11503325 TI - [Cholecystectomy by mini-laparotomy access. Retrospective analysis of the 10-year work]. AB - Ten-year experience of operative treatment of cholelithic disease using mini access cholecystectomy (MACH) in 920 patients was summarized. There were operated on 167 patients for an acute cholecystitis, chronic calculous cholecystitis--716, gall bladder polyposis--36, gall bladder cancer--1. In 48 patients the MACH was matched with choledochus drainage, in 9--choledocholithotomy was performed, in 1- choledochoduodenostomy. In 6 observations the MACH were done simultaneously inguinal herniotomy, in 10--umbilical herniotomy, in 7--uterine extirpation, in 13--operation for ovarial cyst. The bile leakage from gall bladder bed occurred in 3 observations, the operative wound suppuration--in 4. Duration of stationary treatment was 4.1 days at average. PMID- 11503326 TI - [Reoperation for recurrency of dysphagia in patients with cardiospasm and achalasia of cardia]. AB - Results of treatment of 129 patients with dysphagia recurrency after operative treatment of cardiospasm and achalasia of cardia for the 1970-2000 yrs period were analyzed. There were operated 52 (40.3%) of patients, 77--were treated conservatively. The mainly applied methods of operation were those elaborated in the clinic: reoesophagocardiomyotomy with incomplete fundoplication (in 28 observations) and extirpation of the eosophagus with plasty using gastric tube (in 18). Positive result, noted in majority of observations, witnessed the correctness of substantiation of indications and choice of operative intervention in these patients. PMID- 11503327 TI - [Quality of life of patients with colostomy after colonic injury]. AB - The immediate and late follow-up result of surgical treatment of 98 patients with colostoma, formated for the colonic damage, was investigated. The life saving of a survivor constitutes the main task of operating surgeon. The late follow-up period course was not investigated. The method of estimation of the patient quality of life was elaborated and applied for assessment of the medicinal rehabilitational measures efficacy. Unfortunate site of the stoma putting up on anterior abdominal wall, mistakes during its formation, the absence of qualitative stomal rigging, the absence or inadequacy of specialized dispanseric follow-up conduction unfavourably influence the quality of life. When specialized medicinal-rehabilitational measures are applied the quality of life indexes stabilization is observed in patients, to whom colostoma was performed, approximately in 3 months after the damage occurs, persisting during 3-4 months. PMID- 11503328 TI - [Peculiarities of surgical treatment of chylous peritonitis]. AB - Ten own observations of chyloperitoneum for the 1984-2000 yrs period were analyzed. Algorithm of steps of surgeon in revealing chyloperitoneum was substantiated. PMID- 11503329 TI - [Influence of hyperbaric oxygenation on clinico-immunologic indices in children with acute diffuse peritonitis]. AB - The impact of hyperbaric oxygenation (HBO) on clinical-immunologic indexes in complex treatment of an acute diffuse peritonitis of appendicular genesis in children was studied up. Efficacy of the HBO application in moderate immunodepressive state, registered in this disease, was noted. PMID- 11503330 TI - [Surgical treatment of chronic fibrous pancreatitis without ductal hypertension]. AB - Experience of treatment of 36 patients with fibrous chronic pancreatitis without ductal hypertension was summarized. In 15 (41.7%) patients was done operation with direct intervention on pancreas, in 21 (58.3%)--without it. Good and satisfactory result was noted in 94.5% of patients. PMID- 11503331 TI - [Clinical efficacy of alprostan in treatment of obliterating multifocal atherosclerotic affection of arteries in patients with critical ischemia of lower extremities]. AB - Experience of application of alprostan (PGE1, alprostadil) in 19 patients with atherosclerotic affection of vessels, chronic and acute ischemia of lower extremities in III-IV stage in combination with affection of other vascular basins was summarized. In 10 patients, to whom operative vascular reconstruction was contraindicated and/or technically unrealizable, the conservative treatment was conducted using alprostan during 12-25 days up to the clinical effect achievement (total dose up to 1.6 mg). In 9 patients after performance of conservative treatment using alprostan the vascular bed reconstruction was done. Daily infusion of alprostan in 0.1 mg dose diluted in 250-400 ml of isotonic solution of sodium chloride during 2.5-3 h with duration of course not less than 15 days and 1.2-2.2 mg total dose of preparation constitutes an optimal scheme of treatment. Good result was noted in treatment of patients in stage II of ischemia, application of alprostan in stage IV of ischemia had permitted to reduce the extremity amputation volume or to escape it on the whole, and to restore more rapidly the cutaneous integrity in the trophic disorders regions. The alprostan usage is trustworthy in patients with multifocal atherosclerosis with the cardiac coronary vessels affection in an ischemic heart disease. PMID- 11503332 TI - [Clexane and regional anesthesia. Contraindication or possible combination?]. AB - Results of the efficacy and safety analysis of low-molecular weight heparine clexane (sodium enoxaparine) in patients with urological diseases were adduced. Postoperative usage of clexane along with epidural anesthesy had prevented occurrence of hypercoagulation as well as epidural and spinal hematoma. PMID- 11503333 TI - [Blood reinfusion in treatment of thoracic and abdominal trauma]. AB - The experience of application of the blood reinfusion in complex of reanimational measures in treatment of 280 injured persons with thoracoabdominal damage in an acute period of trauma was summarized. Reinfusion was done in 42.8% of operated injured persons, promoting to preserve 243.1 l of preserved blood and simplifying rendering the assistance in an acute period of trauma. PMID- 11503334 TI - [Surgical treatment of patients with consequences of combined trauma of upper extremity complicated by purulent infection]. AB - Result of treatment of 256 patients with consequences of combined trauma of upper extremity was analyzed. In 83 (32.4%) patients the purulent-necrotic complications occurred, caused by mistakes committed on the treatment process phase, especially during rendering the primary aid. It had promoted occurrence of the joint's contracture and ancilosis, osteomyelitis, ischemic contracture and the skin defects as well. Approach to the treatment conduction was in accordance with general principles of the patients orthopedic rehabilitation with consequences of combined trauma of upper extremity. Sanation of purulent focus was performed in 19 patients and microvascular transplantation of tissues--in 13 before restorational procedure was done. PMID- 11503335 TI - [Surgical treatment of intrathoracic compression of respiratory ways caused by mediastinal teratocarcinoma in children]. AB - Twelve children with mediastinal teratodermoid tumor, causing intrathoracic compression of respiratory ways (ITCRW) in 8 of them, were treated in clinic. In 75% of children with ITCRW the conduction of special preoperative preparation was necessary. All the patients were operated on. Tumor was removed en bloc or using the fragmentation method. In secondary tracheomalacia the anterior aortopexy for the tracheal lumen preservation was done simultaneously. PMID- 11503336 TI - [Application of ultrasonic aspiration in surgical treatment of glial tumors]. AB - Results of surgical treatment of 139 patients with cerebral glioma of supratentorial localization were analyzed. The method of ultrasonic aspiration of tumor was applied for the surgical intervention radicalism raising. Application of ultrasonic aspiration had guaranteed the conditions for the controlled excision of tumoral tissue, the hemostasis performance, permitted to reduce cerebral traumatization and to preserve main vessels in the tumor environment, promoted the uncomplicated postoperative period course, rapid reconvalescence of the patients, the neurologic functions preservation, reduction of the patient's invalidization possibility, improvement of his quality of life. PMID- 11503337 TI - [Reaction of peripheric immune organ on action of low-energy red spectrum lasers]. AB - Influence of low-energy laser irradiation of red diapason on the popliteal lymph nodes (LN) structure of sexually matured white rats depending on the given locally irradiation dose, using light conductor of laser apparatus ALOK-1, was investigated. In the irradiated LN the regular dose-dependent quantitative and qualitative changes in parenchyma were observed. In contralateral LN the same, but less pronounced, changes were observed, witnessing about systemic character of peripheral immune organs reactions on the laser irradiation influence. During irradiation in 0.3 and 3 J/cm2 doses the activation of processes in the LN parenchyma, and in 30 J/cm2 dose the destruction processes of LN were observed. PMID- 11503338 TI - [Application of baxtims preparation in treatment of experimental diffuse purulent peritonitis]. AB - The proteins proteolysis increase as well as their biosynthesis disorder were observed in experimental diffuse purulent peritonitis, which was simulated in rats of Wistar line. Application of baxtims preparation for the abdominal cavity treatment during 14 days had promoted restoration of this process and the proteinic deficiency elimination as well. PMID- 11503339 TI - [Reconstruction surgeries in patients after ileostomy]. PMID- 11503340 TI - [Treatment of chronic arterial insufficiency using chemical desympathyzation under control of computer tomography]. PMID- 11503341 TI - [Obturator of external intestinal fistula]. PMID- 11503342 TI - [Non-standard operative interventions in patients with complicated gastroduodenal ulcer concurrent with peritonitis]. PMID- 11503343 TI - [Observation of successful operative treatment of osteal form of Recklinghausen's disease]. PMID- 11503344 TI - [Principles of reconstructive surgery of digestive tract]. PMID- 11503345 TI - Implementation of a medical geographic information system: concepts and uses. AB - This paper introduces a medical geographic information system which has been implemented to enhance public-health research by facilitating the modelling of spatial processes of disease, environment, and healthcare systems in a rural area of Bangladesh. In 1966, a surveillance system was implemented to record all vital demographic events in the study area. Selected information on reproductive and child health, socioeconomic conditions, and health and family-planning interventions is being collected for the surveillance database. This paper discusses the conceptual design of integrating the surveillance database with the medical geographic information system and its use in conducting multidisciplinary health research. The paper is intended to help those who wish to implement a health-based geographic information system to understand the links between people and their environments and to better meet the health needs of target communities. PMID- 11503346 TI - Validity of women's self-reported obstetric complications in rural Ghana. AB - This retrospective study assessed the utility of women's self-reports to identify obstetric complications in rural Ghana. All consenting obstetric and postpartum inpatients, presenting from the seventh month of gestation to 42 days postpartum, were interviewed at the Holy Family Hospital, Techiman and were asked about their signs and symptoms. A combination of clinical examination and laboratory testing of urine and blood samples was used for determining case status. Self-reported obstetric complications of 340 women were compared with the corresponding diagnostic status for their sensitivity, specificity, predictive value, and test efficiency. Using algorithms that could not be practically applied at the community level, self-reported symptoms correctly identified the majority (75%) of complicated and uncomplicated pregnancies, but missed one-quarter of cases requiring emergency obstetric care. The positive predictive value of 50% indicates that women's self-reported symptoms should not be used in estimating the incidence of these conditions or in identifying women requiring referral in this population. PMID- 11503347 TI - Rate of caesarean section as a process indicator of safe-motherhood programmes: the case of Kenya. AB - The study assessed the value of currently-available data on the rates of caesarean section as an indicator of safe-motherhood programmes. Data, collected through the routine health information system of the Ministry of Health, Kenya, were used for analyzing the available process indicators. The methodology of this study illustrates both usefulness and limitations of readily-available healthcare information. The rate of hospital-based caesarean section was 6.3% of all births (range 0.3-37%), whereas the rate of population-based caesarean section was 0.95% (range 0.1%-4%). The rate of population-based caesarean section indicates a significant unmet need for obstetric care in the rural areas and may be a useful tool for monitoring progress on safe-motherhood initiatives in poor settings. Rates of population-based caesarean section are low in Kenya, especially in the rural areas. The rate of caesarean section may be a valuable process indicator for identifying the gaps in obstetric care and may be used for advocating improvements for healthcare to the relevant authorities. PMID- 11503348 TI - Influence of placental malaria infection and maternal hypergammaglobulinaemia on materno-foetal transfer of measles and tetanus antibodies in a rural west African population. AB - Placental malaria infection jeopardizes pregnancy outcome, and its influence may also impair the transplacental transfer of some antibodies. Two hundred and thirteen Gambian mother-baby pairs were studied to determine the influence of placental malaria infection and maternal hypergammaglobulinaemia on transplacental transfer of measles and tetanus antibodies in Gambian population. Placental blood and tissue were collected for placental malaria diagnosis. Cord and maternal sera were tested for total IgG concentration by laser nephelometry and for IgG antibody to tetanus toxoid and measles by ELISA. The prevalence of placental malaria infection was 51.1%. Mothers whose placentae were parasitized had a significantly higher mean total serum IgG (22.0 g/L vs 11.3 g/L, p < 0.001) and measles antibody level (4.02 IU/mL vs 1.21 IU/mL, p < 0.01), but not tetanus antibody, than mothers with non-parasitized placentae. Results of multiple regression analysis showed that placental malaria infection and maternal hypergammaglobulinaemia were associated with the reduction of 72% (95% CI 67.84) and 86% (95% CI 76.91) in transplacental transfer of measles antibody respectively but did not influence the transfer of tetanus antibody. It is concluded that the combined influence of placental malaria infection and maternal hypergammaglobulinaemia is significantly associated with the transfer of impaired measles antibody in this population. PMID- 11503349 TI - Policy of universal salt iodization in Bangladesh: do coastal people benefit? AB - Iodine deficiency disorders (IDD) are a serious public-health problem in Bangladesh and in other countries. Use of iodized salt has been promoted to solve the problem. A study was conducted in eight unions of Chakaria upazila in the Cox's Bazar district of Bangladesh during 1997-1998 to determine the prevalence of use of iodized salt, explore the reasons behind nonuse, and identify the socioeconomic correlates of its use. A quantitative survey was conducted to collect information from 21,190 households on socioeconomic status, demographic characteristics, and the kind of salt used. In-depth interviews and focus-group discussions were also conducted to understand the situation further. The results revealed that only 1.9% of the households used iodized salt. Strong barriers that limit their use of iodized salt included the wide availability of coarse salt, lack of knowledge about the link between iodized salt and IDD, and the high cost of iodized salt. Households in the salt-producing localities and those that are economically disadvantaged tend to use iodized salt less than others. Understanding of the prevalent situation will allow the policy-makers to take measures to improve the situation in the salt-producing areas. PMID- 11503350 TI - Sanitation, community environments, and childhood diarrhoea in rural Zimbabwe. AB - Childhood diarrhoea can be transmitted within the household and community environments. This study examined the effect of partial latrine coverage on diarrhoeal morbidity at the household and community levels. Weekly morbidity data on 272 children, aged less than five years, were collected for 45 weeks from October 1996 to August 1997 in two neighbouring semi-arid communities in rural Zimbabwe. The communities were similar except for sanitation. In Community A, 62% of the children lived in households with a latrine, and in Community B, there was no sanitation. Diarrhoeal morbidity was 68% lower in Community A than in Community B. In addition, the children from the households in Community A without a latrine had lower diarrhoeal morbidity than the children in Community B. The inter-community difference in diarrhoeal morbidity was likely to be due to the latrines and associated hygiene education interrupting intra- and inter-household transmission of diarrhoea. The latrines reduced faecal contamination of the community environment, and as diarrhoeal morbidity fell, contact between infectives and susceptibles within the community would also have fallen. The neighbourhoods in Community A where the adjacent households did not have sanitation had higher diarrhoeal morbidity than in the neighbourhoods where one household did have sanitation. The results of the study suggest that sanitation programmes, which share building costs between the state and individuals, i.e. households, and hence, do not achieve complete coverage in a community, benefit the whole community, because the households that cannot afford a latrine experience lower diarrhoeal morbidity due to the safer community environment. PMID- 11503351 TI - Diarrhoeal morbidity among young children in Eritrea: environmental and socioeconomic determinants. AB - Diarrhoeal diseases are still a major cause of morbidity and mortality among children in many developing countries, particularly in sub-Saharan Africa. In Eritrea, diarrhoeal disease is one of the most frequent causes of illness and a major contributor to mortality among children. Yet, except for some descriptive reports by the National Statistics Office of Eritrea, no systematic study of the factors that influence the prevalence of diarrhoeal morbidity among young children in Eritrea was carried out. This study examined the effects of some environmental and socioeconomic factors that were associated with childhood diarrhoea in Eritrea. The study used data collected by the Demographic and Health Survey (DHS) in Eritrea in 1995. Results of the study showed that the environmental and socioeconomic conditions of the population were significant predictors of diarrhoeal morbidity. The study also found an important association of diarrhoeal morbidity with the age and the number of children, particularly with high prevalence of diarrhoea at the age of weaning and in households with a large number of children. PMID- 11503352 TI - Perceptions of and management practices for Diarrhoeal Diseases by traditional healers in northeastern Nigeria. AB - In Nigeria, there is a paucity of data on the beliefs and practices of traditional healers concerning diarrhoeal diseases. This study was undertaken to provide baseline data for integrating activities of traditional healers into the national Control of Diarrhoeal Diseases (CDD) programme. Interviews of 14 traditional healers, drawn from two large ethnic groups in northeastern Nigeria, were conducted. All but one of them willingly discussed their beliefs and practices. Almost all traditional healers used herbs expecting that it would stop diarrhoea episode. Most traditional healers (n = 11) were, however, aware of the sugar-salt solution (SSS), and six of them had positive impression about it. Ten were willing to be further trained in the preparation/use of SSS, and five reported its past use. Some healers strongly believed that breast-feeding was an important cause of diarrhoea and that, in some cases, diarrhoea was only amenable to traditional treatment. The results of this study suggest that the traditional healers in the region may contribute to promoting the appropriate home management of diarrhoea. However, their perceptions and practices need to be upgraded to ensure success. PMID- 11503353 TI - Research to support household and community IMCI. Report of a meeting, 22-24 January 2001, Baltimore, Maryland, USA. AB - The Integrated Management of Childhood Illness (IMCI) strategy combines improved case management of childhood illness with aspects of nutrition, immunization, disease prevention, and promotion of growth and development. The household and community component of IMCI was formulated to reach the numerous sick children who are ill and often die at home without ever being treated by a trained healthcare practitioner. In January 2001, USAID (Child Health Research Project and BASICS II) and the CORE Group sponsored a meeting in Baltimore, Maryland, to determine the research needed to implement household and community IMCI effectively. This paper summarizes the presentations at that meeting and highlights the research and programme priorities expressed using the three element approach devised by the CORE Group and USAID (BASICS II and Child Survival Technical Support Project). Research priorities to improve partnerships between health facilities and the communities they serve (Element 1) include finding ways to increase community involvement and management of health facilities, establishing accurate costs for community IMCI services, and formulating cost-recovery mechanisms tailored to local circumstances. Programme priorities in Element 1 include establishment of systems for maintenance of an adequate supply of essential equipment and medicines, while retaining access for the poor and ensuring adequate referral mechanisms for severely-ill patients that include monitoring and incentives for the performance of health workers. Research priorities to increase appropriate, accessible care and information from community-based care providers (Element 2) consist of activities to design simplified IMCI guidelines for use by community health workers (CHWs) and volunteers and evaluation of the impact of using these guidelines on morbidity and mortality. Also a priority item in this category is experimenting with ways to teach mothers and families to care for sick children at home and strategies to improve the practice of medicine in the private sector. More research is also needed to improve the quality of nutritional and preventive health counselling given by CHWs and to find ways to make community IMCI interventions sustainable. Programme priorities in Element 2 include efforts at the national level to establish policies to improve care by traditional and private care providers and to define the position of volunteer workers in the national health system. Research priorities to integrate promotion of key family practices critical for public health (Element 3) are focused on determining which interventions are the most effective in reducing child morbidity and mortality at the household and community levels, finding the best methods of delivering these interventions, and implementing these and scaling up in essential service packages. Particular issues in child health, needing urgent attention from the research and programme communities, are HIV/AIDS and neonatal morbidity and mortality. PMID- 11503354 TI - Chest conduction properties and ECG equalization. AB - In common practice of detecting and recording biomedical signals, it is often implicitly assumed that the propagation, through the whole circuit human body electrodes recording devices, is frequency and voltage independent. As a consequence, clinicians are not aware that recorded signals do not correspond faithfully to the original electrical activity of organs under investigation. We have studied the transmission of electrical signals in human body at various voltages and frequencies to understand if and to which extent the most diffused stimulating and recording techniques used in medicine are affected by global body conduction properties. Our results show that, in order to obtain a more faithful detection of electrical activity produced or evoked by human organs (e.g. EGG, electromyography, etc.), it is convenient to 'equalize'' recorded signals. To this purpose, two equalization techniques are proposed, based, respectively, on a simple hardware filtering during acquisition, or FFT post-processing of the acquired signals. As an application, we have studied the transmission of electrical signal in human chest and have compared equalized high frequency ECG signals with raw (original) recordings. PMID- 11503355 TI - A cross-sectional study of workers with occupational exposure to petroleum derivatives. AB - Expression of human involucrin (hINV) a protein of the cornified cell envelope, was studied in the skin of gasoline storage workers, in order to evaluate the effects of the exposure to petroleum derivatives. A total of 25 forearm skin punch biopsies were carried out. Twenty of which were performed on exposed subjects and five on controls. The specimens were processed for immunohistochemistry and hINV expression was evaluated using an anti-hINV monoclonal antibody and the ABC technique. Percentage of immunolabeled keratinocytes was significantly higher in subjects exposed to gasoline with respect to the control sample. A premature hINV expression was detected both in suprapapillary and interpapillary keratinocytes. Such overexpression of hINV seems to be related to an attempt of increasing skin defence mechanism. Therefore it was concluded that also in absence of clinical skin manifestation the exposure to gasoline determines an involvement of keratinocytes on molecular basis. PMID- 11503356 TI - [Eulogy for Jean-Charles Sournia (1917-2000)]. PMID- 11503357 TI - [Prevention of congenital toxoplasmosis in France. Risk assessment. Results and perspectives of prenatal screening and newborn follow up]. AB - In France, a national program for the prevention of congenital toxoplasmosis has been set up 25 years ago. This program is here presented and discussed in details. It is based on a decision tree well defined, with pre and/or per gravidic serological screening with several different tests, completed, if necessary, by ultrasounds examinations of the fetus, biomolecular tests (PCR) on amniotic fluid, and by clinical, biological, and radiological surveillance of neo nates. The purpose of this prevention program is to: 1/identify nonimmune young women and limit their contamination risk during pregnancy by appropriate counseling on hygiene and diet; 2/screen and treat per gravidic toxoplasmosis as early as possible so as to prevent or limit transmission to the fetus and its consequences. 3/in utero diagnose and treat infestation of the fetus; 4/diagnose and treat asymptomatic congenital toxoplasmosis in neonates, to prevent risks of reactivation and late complications, especially ocular. Such a prevention program has a cost validated by the prevalence of acquired toxoplasmosis in adults in France (over 50% of the population) and by the yearly incidence of congenital toxoplasmosis (at least 0.1% of births according to the best hypothesis). These 6 to 700 congenital toxoplasmosis cases per year may be compared to the 6 to 7,000 per gravidic seroconversions which could lead to fetal contamination if no preventive measures are taken. Nevertheless, as it is often the case in the field of prevention, it is very difficult to statistically assess the efficacy of this program even though several arguments show that it allows to eliminate the most serious toxoplasmosis, sources of serious handicaps at birth, and to limit the frequency of late complications (especially retino-choroiditis) of asymptomatic infections in neonates. The position of European countries varies as to prevention of congenital toxoplasmosis. Some countries (Austria, Belgium) have national prevention programs similar to the French one, whereas others have set up only limited programs or set up no systematic prevention. These differences may be accounted for by the different frequencies of toxoplasmic risk. It seems mandatory to forget all dogmatism and not to stick to a strictly statistical approach for a disease with not only medical but also social and human consequences. PMID- 11503358 TI - [Neuronal connectivity and chemical mediators involved in olfactory message transmission]. AB - In this review, we discuss some of the neural processes involved in the perception of odors which, together with audition and vision, provide essential information for analyzing our surroundings. We shall see how odor detection and learning induce substantial structural and functional changes at the first relay of the olfactory system, i.e., the main olfactory bulb. Among the mechanisms which participate in these modifications are changes in the cell's responses to a transmitter and the persistence of a high level of interneuron neurogenesis within the adult olfactory bulb. Our goal is to present some observations related to these two phenomena that may aid in understanding the neural mechanisms of sensory perception and shed light on the cellular basis of olfactory learning. To this purpose, we summarize the current ideas concerning the molecular mechanisms and organizational strategies used by the olfactory system to transduce, encode, and process information at various levels in the olfactory sensory pathway. Due to space constraints, this review focuses exclusively on the olfactory systems of vertebrates and primarily those of mammals. PMID- 11503359 TI - [New functional cerebral cartography: studies of plasticity of the human brain]. AB - The non-invasive brain scanning techniques, introduced a quarter of a century ago, have become crucial for diagnosis in clinical neurology. They have also been used to investigate brain function and have provided information about normal activity and pathogenesis. They have been used to investigate functional specialisation in the brain and how specialised areas communicate to generate complex integrated functions such as speech, memory, the emotions and so on. Brain plasticity is a generic term that has now come to be used to signify any changes in brain structure or function. The phenomenon is poorly understood, and yet clinical neurologists are aware that spontaneous recovery from brain lesions is not uncommon. An improved understanding of the mechanisms of recovery may generate new therapeutic strategies and indicate ways of modulating mechanisms that promote plastic compensation for loss of function. The main methods used to investigate these issues are positron emission tomography and magnetic resonance imaging (MRI). The techniques of functional brain mapping and computational morphometrics depend on high performance scanners and a validated set of analytic statistical procedures that generate reproducible data and meaningful inferences from them. The motor system presents a good paradigm to illustrate advances made by scanning towards an understanding of plasticity at the level of brain areas. The normal motor system is organised in a nested hierarchy. Recovery from paralysis caused by internal capsule strokes involves functional reorganisation manifesting as changed patterns of activity in the component brain areas of the normal motor system. The pattern of plastic modification depends in part on patterns of residual or disturbed connectivity after brain injury. Therapeutic manipulations in patients with Parkinson's disease using deep brain stimulation, dopaminergic agents or foetal mesencephalic transplantation provide a means to examine mechanisms underpinning plastic change. Other models of plastic change, such as normal visuospatial learning or re-establishing speech comprehension after cochlear implantation in the deaf illustrate how patterns of brain function adapt over time. Limitations of the scanning techniques and prospects for the future are discussed in relation to new developments in the neuroimaging field. PMID- 11503360 TI - [Compulsory medical certificates of early childhood: unknown epidemiology tool]. AB - The delivery of three medical certificates has been made compulsory in 1970, the first during the first eight days of life, the second and the third respectively at 9 and 24 months. Thirty years later a survey has been conducted to evaluate the application and the efficiency of the system in 100 french departments. All the medical directors of the departmental MCH service send back the questionnaire. The number of annual birth by department is of 7,665 (mean) with extremities of 800 to 36,457. The covering of the concerned population is very good for the first certificate and increasing from 88.8% in 1992 to 94.9% in 1998; it is less good and not increasing for the CS9 (71.1%) and the CS24 (66%). The answers to each item are also variable according to the department but with an average of 86% (to refer to 650,000 annual births). The computer treatment of the certificates is increasing since ten years but 8 departments are still using manual analysis. In spite of the decentralization of MCH service each department is obliged to send to the Ministry of Health, each year, the results of 22 indicators which makes it possible to follow up the evolution of the situation of young children at the national level and to establish comparasons between departments. Simultaneously the certificates allow to guide the MCH activities, often at the canton level. The first certificate has been very useful for the study of perinatal problems and the third for the vaccination coverage. For example, some results are given and discussed (qualification of the examiner, neonatal mortality and socio-economical categories, mother age, duration of pregnancy, birth weight, vaccination coverage...). Of course, the system is not perfect but is improving due to a better information of the medical personnel and of the families. It is still necessary to continue the research to underline the deficiencies and to correct them. Probably, in the future, the system will be extended to the registration of data obtained at the maternal school (3-4 years old) and the elementary school (5-6 years old). PMID- 11503361 TI - [Members of the Vaccine Central Committee: a group of men deserving their native land and even humanity]. AB - Two hundred years ago, in May 1800, a private initiative set up a "Central Committee of Vaccine" in Paris. A handful of men launched an extraordinary research: how to implement a still recent and little-known discovery, a method to protect people against small pox. The news came from England, with which France was on bad terms. Smallpox was a fearsome disease killing one tenth of the population and disfiguring or maiming as many again. After several failed attempts, success was reached and the value of the method demonstrated. The committee played a major role in spreading the vaccine (thanks to Jenner) not only in France but also in the whole of the Napoleonic Empire. His remarkable experiments were published and made known to the whole western world. The Committee was made official en 1804 and operated until the foundation of the Academy of medicine, which took over its duties and responsibilities. The French owe a lot to this Central Committee of Vaccine, which greatly contributed to fighting small pox and eradicating the disease finally. PMID- 11503362 TI - [The beginnings of vaccine diffusion in France (1800-1850)]. AB - First factor of mortality at the eighteenth century, smallpox killed each year about 50.000 to 80.000 people in France and 25.000 to 30.000 in England. In 1796, Edward Jenner discovered the fabulous properties of the cowpox which, transplanted from cow to human, immunized against the disease. In France, between 1800 and 1850, a few hundreds of vaccinators took part in a significant crusade against smallpox. They went in the villages and the thatched cottages, fought against the routine, and, sometimes, against the hostility of the mayors or the priests. Moreover, the cow-pox often missed or lost its strength. Despite everything, their efforts were crowned success: during the nineteenth century, small-pox mortality drops by 90%. PMID- 11503363 TI - [Eradication of infectious diseases and vaccination]. AB - Immunizations in man may act upon the infectious diseases development in three evolutive patterns: eradication, elimination, or disease control. Since the eradication of smallpox, very few diseases are at present candidates for a next eradication; poliomyelitis will be the first, perhaps followed by measles in many years. In spite of efficacious vaccines and a solely human reservoir, the eradication requires very strict conditions. However some diseases have been recently successfully eliminated: poliomyelitis (Americas, Europe, West Pacific regions), measles (i.e. Finland, United States), rubella (i.e. Finland). In spite of some limits, diphtheria, pertussis, hepatitis B are well controlled providing to get a sufficient vaccine coverage and to improve a sustained surveillance of those diseases. Regarding Haemophilus influenzae b, infections it is premature to predict a near elimination and the control is not yet well defined. National and regional programs have often to improve immunisation coverage and a better surveillance. PMID- 11503364 TI - [Vaccines outside of Europe. Light and shadow of a victory]. AB - In the beginning of the 19th century, vaccination against smallpox spread rapidly in Europe and out of Europe. Although most statesmen eagerly adopted the vaccine, populations displayed mixed attitudes, ranging from mild enthusiasm to resistance. The article illustrates the social response with examples drawn from Algeria, Tunisia, Egypt and Brazil. This analysis puts in sharp focus the crucial importance of a factor for the managers of public health: the adhesion of populations. PMID- 11503366 TI - Field study of dairy cows with reduced appetite in early lactation: clinical examinations, blood and rumen fluid analyses. AB - The study included 125 cows with reduced appetite and with clinical signs interpreted by the owner as indicating bovine ketosis 6 to 75 days postpartum. Almost all of the cows were given concentrates 2 to 3 times daily. With a practitioners view to treatment and prophylaxis the cows were divided into 5 diagnostic groups on the basis of thorough clinical examination, milk ketotest, decreased protozoal activity and concentrations, increased methylene blue reduction time, and increased liver parameters: ketosis (n = 32), indigestion (n = 26), combined ketosis and indigestion (n = 29), liver disease combined with ketosis, indigestion, or both (n = 15), and no specific diagnosis (n = 17). Three cows with traumatic reticuloperitonitis and 3 with abomasal displacement were not grouped. Nonparametric methods were used when groups were compared. Aspartate aminotransferase, glutamate dehydrogenase, gamma-glutamyl transferase and total bilirubin were elevated in the group with liver disease. Free fatty acids were significantly elevated in cows with ketosis, compared with cows with indigestion. Activity and concentrations of large and small protozoas were reduced, and methylene blue reduction time was increased in cows with indigestion. The rumen fluid pH was the same for groups of cows with and without indigestion. Prolonged reduced appetite before examination could have led to misclassification. Without careful interpretation of the milk ketotest, many cases with additional diagnoses would have been reported as primary ketosis. Thorough clinical examination together with feasible rumen fluid examination and economically reasonable blood biochemistry did not uncover the reason(s) for reduced appetite in 14% of the cows. More powerful diagnostic methods are needed. PMID- 11503365 TI - In vitro evaluation of frozen-thawed stallion semen: a review. AB - The article reviews methods used for in vitro evaluation of sperm, with particular emphasis on frozen-thawed stallion sperm. The techniques, limitations of the methods and correlations with fertility results are discussed. Very few studies have tried to find correlation between fertility of frozen stallion semen and laboratory tests. It is difficult and expensive to inseminate an adequate number of mares to achieve statistically significant differences. Significant, but low correlations have been demonstrated between the foaling rate and subjective motility of sperm incubated for 2 h and 4 h at 37 degrees C and hypoosmotic swelling test after 0 and 3 h of incubation. Significant correlations have been reported between the pregnancy rate and viability of propidium iodide stained sperm assessed by flow cytometry as well as for glass wool and Sephadex filtration tests. No correlations have been detected between fertility and motility immediately after thawing. In spite of that, motility estimation by light microscope is the most commonly used method to evaluate frozen-thawed stallion sperm. Computer assisted automatic sperm analyzers have replaced light microscopy in research projects, but so far nobody has been able to demonstrate a correlation between fertility of frozen stallion semen and any of the motility parameters obtained by these instruments. PMID- 11503367 TI - Calcification of the intervertebral discs and curvature of the radius and ulna: a radiographic survey of Finnish miniature dachshunds. AB - The vertebral column of 124 randomly selected miniature dachshunds, representing 4.5% of the population registered by the Finnish Kennel Club during the years 1988 to 1996, were radiographed. The front legs were also radiographed in order to evaluate the curvature of the radius and ulna. Calcified discs were found in 75.9% of the longhaired miniature dachshunds and in 86.7% of the wirehaired ones. The occurrence of signs associated with IDD was 16.5% in longhaired and 15.6% in wirehaired miniature dachshunds. The occurrence of signs of IDD in dogs with calcified discs was 20.0% and 17.9% in longhaired and wirehaired miniature dachshunds, respectively. In dogs without calcifications only one dog showed signs of IDD. The curvature of the radius and the ulna did not differ between the dogs with signs of IDD and the healthy ones, or between the dogs with and without intervertebral calcifications. Our results indicate that radiographic eradication based on the presence of intervertebral calcifications is not suitable for breeding purposes for the Finnish miniature dachshund population because the percentage of dogs without calcifications is small. PMID- 11503368 TI - Specific immune response of mares and their newborn foals to Actinobacillus spp. present in the oral cavity. AB - Oral swab samples, serum and colostrum was taken from 15 mares and 14 of their foals, within 24 h of birth. The presence of antibody against Actinobacillus spp. isolated from the oral cavity was investigated using agar gel immunodiffusion. Antibodies against 48 out of the 77 Actinobacillus isolates from all horses in the study were present in the respective sera of 13 mares and 9 foals. In 11 mother-foal pairs, the antibody content of the foal serum was similar to that of the mare, and in 9 cases this was reflected in the antibody content of colostrum from the mare. The results indicate that an immune response to Actinobacillus spp. colonising the oral cavity is present in many adult horses and that this immune response can be transferred from mother to foal via colostrum. PMID- 11503369 TI - Effect of tetracosactid on post partum cyclicity in cows after induction of parturition with PGF2 alpha. AB - Parturition and retention of fetal membranes were induced with PGF2 alpha in 3 primiparous dairy cows. Starting on day 12 post partum (PP) the cows were treated with 500 micrograms i.m. of ACTH-analogue (tetracosactid) every 6 h for 6 times. Changes in plasma concentrations of cortisol, progesterone and 15-ketodihydro PGF2 alpha were evaluated immediately after treatment. The effects on the resumption of ovarian activity were evaluated by clinical and ultrasound examinations and by progesterone and 15-ketodihydro-PGF2 alpha analyses for 56 days after parturition. Treatment was able to induce a statistically significant (p < 0.01) similar increase in cortisol and progesterone after both the 1st and the 6th injections, in all cows. No changes in 15-ketodihydro-PGF2 alpha concentrations were seen after any of the injections of ACTH-analogue. The first corpus luteum (CL) was seen on day 18 PP (cow A), and 28 (cow B) and in both cases it was followed by a normal ovarian cyclicity. No CL was observed during the whole period of study in cow C. Progesterone profiles confirmed these clinical and ultrasonographic findings. The steroid output, especially progesterone, induced by the ACTH-analogue might be a stimulus for the onset of ovarian cyclicity, since 2 of the 3 animals ovulated earlier than expected. These findings point to the fact that interference with the stress system might have a positive effect on ovarian cyclicity. The different pattern of response does however demand further studies. PMID- 11503371 TI - Electrolyte composition of mink (Mustela vison) erythrocytes and active cation transporters of the cell membrane. AB - Red blood cells from mink (Mustela vison) were characterized with respect to their electrolyte content and their cell membranes with respect to enzymatic activity for cation transport. The intra- and extracellular concentrations of Na+, K+, Cl-, Ca2+ and Mg2+ were determined in erythrocytes and plasma, respectively. Plasma and red cell water content was determined, and molal electrolyte concentrations were calculated. Red cells from male adult mink appeared to be of the low-K+, high-Na+ type as seen in other carnivorous species. The intracellular K+ concentration is slightly higher than the extracellular one and the plasma-to-cell chemical gradient for Na+ is weak, though even the molal concentrations may differ significantly. Consistent with the high intracellular Na+ and low K+ concentrations, a very low or no ouabain-sensitive Na+, K(+) ATPase activity and no K(+)-activated pNPPase activity were found in the plasma membrane fraction from red cells. The Cl- and Mg2+ concentrations expressed per liter cell water were significantly higher in red cells than in plasma whereas the opposite was the case with Ca2+. The distribution of Cl- thus does not seem compatible with an inside-negative membrane potential in mink erythrocytes. In spite of a steep calcium gradient across the red cell membrane, neither a calmodulin-activated Ca(2+)-ATPase activity nor an ATP-activated Ca(2+)-pNPPase activity were detectable in the plasma membrane fraction. The origin of a supposed primary Ca2+ gradient for sustaining of osmotic balance thus seems uncertain. PMID- 11503370 TI - Plasma calcium, inorganic phosphate and magnesium during hypocalcaemia induced by a standardized EDTA infusion in cows. AB - The intravenous Na2EDTA infusion technique allows effective specific chelation of circulating Ca2+ leading to a progressive hypocalcaemia. Methods previously used were not described in detail and results obtained by monitoring total and free ionic calcium were not comparable due to differences in sampling and analysis. This paper describes a standardized EDTA infusion technique that allowed comparison of the response of calcium, phosphorus and magnesium between 2 groups of experimental cows. The concentration of the Na2EDTA solution was 0.134 mol/l and the flow rate was standardized at 1.2 ml/kg per hour. Involuntary recumbency occurred when ionised calcium dropped to 0.39-0.52 mmol/l due to chelation. An initial fast drop of ionized calcium was observed during the first 20 min of infusion followed by a fluctuation leading to a further drop until recumbency. Pre-infusion [Ca2+] between tests does not correlate with the amount of EDTA required to induce involuntary recumbence. Total calcium concentration measured by atomic absorption remained almost constant during the first 100 min of infusion but declined gradually when the infusion was prolonged. The concentration of inorganic phosphate declined gradually in a fluctuating manner until recumbency. Magnesium concentration remained constant during infusion. Such electrolyte responses during infusion were comparable to those in spontaneous milk fever. The standardized infusion technique might be useful in future experimental studies. PMID- 11503372 TI - Serum calcium response following oral zinc oxide administrations in dairy cows. AB - Six non-pregnant cows were allocated into 3 groups. Group 1 comprised a pair of lactating cows, whereas groups 2 and 3 each comprised a pair of non-lactating cows. The cows in groups 1 and 2 were dosed intraruminally by stomach tube with zinc oxide at 120 mg Zn per kg of bodyweight at weekly intervals for a period of 33 days. Each cow received a total of 4 doses of zinc oxide. Group 3 served as non-treated control group. Blood samples were collected from all 6 cows daily. Serum was analysed for concentration of calcium. Within 12-24 h of each zinc oxide administration the serum calcium of the lactating cows dropped dramatically indicating the existence of an antagonistic effect between Zn and Ca. The first Zn induced hypocalcaemic episode in the lactating cows was followed by a rise in serum calcium to a level above the pre-dosing level and above the mean value of the control group. The depth of the hypocalcaemic response decreased with the number of zinc oxide dosings. This effect was explained as a response from the stimulation of the calcium homeostatic mechanisms. In the Zn dosed non-lactating cows responses were similar but less clear. The perspective of these findings is discussed in relation to resistance towards parturient hypocalcaemia. PMID- 11503373 TI - Optimization of the agar-gel method for isolation of migrating Ascaris suum larvae from the liver and lungs of pigs. AB - Experiments on use of an agar-gel method for recovery of migrating Ascaris suum larvae from the liver and lungs of pigs were conducted to obtain fast standardized methods. Subsamples of blended tissues of pig liver and lungs were mixed with agar to a final concentration of 1% agar and the larvae allowed to migrate out of the agar-gel into 0.9% NaCl at 38 degrees C. The results showed that within 3 h more than 88% of the recoverable larvae migrated out of the liver agar-gel and more than 83% of the obtained larvae migrated out of the lung agar gel. The larvae were subsequently available in a very clean suspension which reduced the sample counting time. Blending the liver for 60 sec in a commercial blender showed significantly higher larvae recovery than blending for 30 sec. Addition of gentamycin to reduce bacterial growth during incubation, glucose to increase larval motility during migration or ice to increase sedimentation of migrated larvae did not influence larvae recovery significantly. PMID- 11503374 TI - Plasma melatonin levels in relation to the light-dark cycle and parental background in domestic pigs. AB - To study porcine melatonin secretion in a stable environment 3 daytime (10.00 15.00) and 3 nighttime (22.00-03.00) plasma samples were collected by jugular venipuncture from 15 gilts, 16 sows, 3 boars and 48 piglets (24 females and 24 males from 8 litters) and analysed for melatonin content. Nighttime melatonin concentrations were higher than daytime melatonin concentrations (p < 0.001), whereas no effect of sampling order could be discerned. The 3 adult Hampshire boars had higher melatonin concentrations during the day and the night, than the 31 adult Yorkshire females (p < 0.05). There was no clear difference between gilts and sows in plasma melatonin. The gilts from one of the litters had higher plasma melatonin concentrations than the gilts in 3 other litters (p < 0.05). Among the 48 piglets, the increase of nocturnal melatonin secretion differed between litters (p < 0.01), whereas the influence of father was not quite significant (p = 0.12). No difference in daytime melatonin concentrations between litters could be found, and there was no difference in melatonin levels between the male and female piglets. In conclusion, this study demonstrates that domestic pigs express a nocturnal increase of melatonin secretion in a standard stable environment. For some animals the amplitude of nighttime melatonin secretion was very low, although always higher than the daytime base levels. Furthermore, the levels of nighttime melatonin secretion differed between litters, which suggests a genetic background. PMID- 11503375 TI - Salmonella dublin in Danish dairy herds: frequency of change to positive serological status in bulk tank milk ELISA in relation to serostatus of neighbouring farms. AB - Bulk tank milk from 1,429 herds were collected in 3 rounds from 19 different geographic areas. The milk samples were tested by use of indirect LPS-ELISA procedure to detect Salmonella dublin antibodies. From the obtained OD-values herd seroprevalence in the given area was determined and GR-scores calculated for each herd by addition of the number of positive sampling rounds by the 5 geographically closest neighbour herds. In the 19 different areas the calculated prevalence ranged from 0.01 to 0.41. Totally 3,697 GR-scores were given. The mean GR-scores in the areas ranged from 0.0 to 6.5. Higher GR-scores were found in herds changing to seropositive status compared with herds seronegative throughout the study period. The results indicate that the risk for a dairy herd to receive S. dublin infection increases with the disease status among the nearest neighbours and with the prevalence of seropositive herds in the geographic area. PMID- 11503376 TI - Experimental infection of conventional pigs with Streptococcus suis serotype 2 by aerosolic exposure. PMID- 11503377 TI - Clinical and endocrine investigations after dexamethasone and prostaglandin induced premature parturition--a case report. PMID- 11503378 TI - Type 2 diabetes. Thoughts about an epidemic and your role in battling it. PMID- 11503379 TI - This week's episode of diabetic law and order. PMID- 11503380 TI - Painful lump on a middle-aged scalp. Squamous cell carcinoma. PMID- 11503381 TI - The hidden patient. Identifying the needs of caregivers. PMID- 11503382 TI - Health care for a legion of aging baby boomers. PMID- 11503383 TI - Alzheimer's disease: the possibility of prevention and early treatment. PMID- 11503384 TI - How does menopause alter the primary care of women? PMID- 11503385 TI - The prospect of "staying nine". PMID- 11503387 TI - Here comes tomorrow. Advances in cardiovascular medicine. PMID- 11503386 TI - "Elder" physician assistants and their practices. AB - Physician assistants (PAs) in American medicine are aging. After 30 years of having PAs enter the US health care system, it seems an appropriate time to examine where they are in their careers. This study describes the phenomenon of a rising number of elder PAs in the ranks of the nation's health providers and their patterns of practice. METHOD: A subpopulation cohort of practicing PAs 60 years and older was compared to all PAs. Age range, gender, practice setting, specialty, and other demographic variables were analyzed using established methods. Differences between elder PAs and all other PAs and services provided by the two groups were examined. RESULTS: Elder PAs constituted about 1.6% of full time PAs in practice in 1998. Average age of elder PAs is 64 years (range, 60 years to 74 years) compared to 41 years (range, 21 years to 60 years) for all full-time practicing PAs. Analysis of several variables--gender, ethnicity, geographic distribution, number of patients seen in a week, and hours worked in a week--revealed few differences between the cohorts. On average, older PAs were in practice longer than all PAs (18.3 years and 9.2 years, respectively), had been with their current employer for 7.7 years, and worked one job. Elder PAs had an associate degree at twice the rate of all PAs, but three times as many of them had a doctorate. Elder PAs are more likely to be taking care of older patients, working in a rural area, and remaining in a rural area longer than PAs overall. INTERPRETATION: America's corps of PAs continues to work into and beyond the usual retirement age for most workers, and is caring for a greater percentage of older persons than all PAs are. Most elder PAs practice in family or general medicine and prefer to practice in a rural area. These practice preferences mirror results of previous studies of the practices of elder physicians in the United States and Canada. The aging of PAs has important implications for clinical practice and health workforce planning. If PAs were encouraged to take early retirement, more elderly patients would be shifted to younger providers- with the potential to affect delivery of primary care services in rural communities. PMID- 11503388 TI - Childhood immunization: schedule for 2000. PMID- 11503390 TI - More on a title change for PAs. PMID- 11503389 TI - Help for new graduates seeking a job. PMID- 11503391 TI - Answering big questions, and making hard decisions. PMID- 11503392 TI - Answering big questions, and making hard decisions. PMID- 11503393 TI - Pharmacists are on the health care team, too. PMID- 11503394 TI - Tests for PAs in specialty practice. PMID- 11503395 TI - In defense of surgical PAs. PMID- 11503396 TI - In defense of surgical PAs. PMID- 11503397 TI - In defense of surgical PAs. PMID- 11503398 TI - Revealing one's true self in a medical journal. PMID- 11503399 TI - Revealing one's true self in a medical journal. PMID- 11503400 TI - Revealing one's true self in a medical journal. PMID- 11503401 TI - Advice on obtaining the valuable Medicare number. PMID- 11503402 TI - Determining the cause and relieving the discomfort of rhinosinusitis. PMID- 11503403 TI - Better care for patients with an allergic disorder. PMID- 11503404 TI - Medicine versus the Medicare purse. Paying the price for prescription drugs. PMID- 11503405 TI - Dangers, myths, controversy. Sublingual nifedipine for hypertensive crisis. PMID- 11503406 TI - Choosing treatment for a PFO-related neurologic event. PMID- 11503407 TI - Physician assistants and the practice of neurology. AB - This is the first study of how physician assistants (PAs) function in neurology practice. Forty-six PAs who work full-time in a neurology practice were surveyed by telephone and by mail; 100% responded to all telephone survey questions on aspects of practice, and almost all responded to the mailed survey. Of these, 26 worked in a group practice; the average duration of neurology experience was 7 years. Diagnoses seen by these PAs were similar to those seen in most neurology practices. The scope of findings is limited but suggests that, in neurology settings where PAs are employed, they perform a broad range of functions. Furthermore, PAs in most neurology practices likely assist in off-loading patients whose requirements for care are considered "routine" by the PAs' supervising physician(s). PMID- 11503408 TI - Argyria: consequences of using an antiquated medication. PMID- 11503409 TI - Survived by a daughter. PMID- 11503410 TI - Hantavirus update on an emerging--and lethal--infectious disease. PMID- 11503411 TI - Evidence-based medicine in everyday practice. Are your decisions based on solid evidence? PMID- 11503412 TI - CT versus plain films in the diagnosis of intestinal obstruction. PMID- 11503413 TI - Proposing centers of excellence in PA studies. PMID- 11503414 TI - Update on migraine management. PMID- 11503415 TI - Examining referral practices of primary care physician assistants. AB - BACKGROUND: Various barriers to specialty referrals that are initiated by physician assistants (PAs) have been reported to limit a PA's ability to care for patients effectively when they have a medical problem that requires specialty intervention. To assist in evaluating this matter, we conducted a survey to assess referral practices and perceived barriers to referral among primary care PAs in the United States. METHODS: A cross-sectional, random sample of 500 primary care PAs from across the United States was surveyed to determine their referral practices and perceived barriers to referrals. Standard descriptive statistics and univariate analysis of variables were carried out using Pearson chi-squared tests. RESULTS: Seventy-one percent of respondents identified barriers to referral of patients to specialists, although 86% were satisfied with their level of autonomy. The most frequently identified barrier (38%) was the patient's insurance company. This is not an unexpected finding, considering the managed care environment in which medicine is practiced in the United States. Of respondents who mentioned this barrier, the majority were in practice 5 years or less (P < .05) and lived in a community with 10,000 or more people (P < .05). Seventeen percent of respondents also identified refusal or reluctance of specialists to accept referrals from PAs as a barrier; of respondents who mentioned this barrier, the majority had been in practice 5 or fewer years (P < .05) and worked in a community of 10,000 or more people (P < .01). A reassuring finding was that the majority of PAs (86%) were satisfied with their level of autonomy in making referrals. This may reflect the level of confidence that supervising and specialty physicians have in a PA's ability to make appropriate decisions regarding referrals. CONCLUSIONS: Further research is needed to determine whether these barriers affect a patient's access to cost-effective, high-quality care from specialist physicians. PMID- 11503416 TI - The growing presence of complementary and alternative medicine. PMID- 11503417 TI - A review of the neurotoxicity risk of selected hydrocarbon fuels. AB - Over 1.3 million civilian and military personnel are occupationally exposed to hydrocarbon fuels, emphasizing gasoline, jet fuel, diesel fuel, or kerosene. These exposures may occur acutely or chronically to raw fuel, vapor, aerosol, or fuel combustion exhaust by dermal, respiratory inhalation, or oral ingestion routes, and commonly occur concurrently with exposure to other chemicals and stressors. Hydrocarbon fuels are complex mixtures of 150-260+ aliphatic and aromatic hydrocarbon compounds containing varying concentrations of potential neurotoxicants including benzene, n-hexane, toluene, xylenes, naphthalene, and certain n-C9-C12 fractions (n-propylbenzene, trimethylbenzene isomers). Due to their natural petroleum base, the chemical composition of different hydrocarbon fuels is not defined, and the fuels are classified according to broad performance criteria such as flash and boiling points, complicating toxicological comparisons. While hydrocarbon fuel exposures occur typically at concentrations below permissible exposure limits for their constituent chemicals, it is unknown whether additive or synergistic interactions may result in unpredicted neurotoxicity. The inclusion of up to six performance additives in existing fuel formulations presents additional neurotoxicity challenge. Additionally, exposures to hydrocarbon fuels, typically with minimal respiratory or dermal protection, range from weekly fueling of personal automobiles to waist-deep immersion of personnel in raw fuel during maintenance of aircraft fuel tanks. Occupational exposures may occur on a near daily basis for from several months to over 20 yr. A number of published studies have reported acute or persisting neurotoxic effects from acute, subchronic, or chronic exposure of humans or animals to hydrocarbon fuels, or to certain constituent chemicals of these fuels. This review summarizes human and animal studies of hydrocarbon fuel-induced neurotoxicity and neurobehavioral consequences. It is hoped that this review will support ongoing attempts to review and possibly revise exposure standards for hydrocarbon fuels. PMID- 11503419 TI - Methyl bromide risk characterization in California. PMID- 11503418 TI - Biomarkers of apoptosis: release of cytochrome c, activation of caspase-3, induction of 8-hydroxy-2'-deoxyguanosine, increased 3-nitrotyrosine, and alteration of p53 gene. AB - Biomarkers rely on biochemical, histological, morphological, and physiological changes in whole organisms. Their use is becoming an important tool to examine changes at cellular and molecular levels, especially in nucleic acids and proteins. Biomarkers are used to measure exposure to a toxic agent, to detect severity of any toxic response, and to predict the possible outcome. Information on the mechanisms of action of toxicants can allow the development of potential biomarkers of effect and thus improvement of the risk assessment processes. Use of biomarkers as a tool to predict induction of apoptosis allows identification of biological signs that may indicate increased risk for disease. In cells undergoing apoptosis, the release of cytochrome c from the mitochondria to the cytoplasm and the activation of caspase-3, a key enzyme to execution stage of apoptotic pathway, have been studied as biomarkers of cell death (apoptosis). Products of DNA fragmentation that either accumulate in the cellular tissues or are excreted in the urine are useful markers of DNA damage. The induction level of urinary or cellular level of 8-hydroxy-2-deoxyguanosine and 3-nitrotyrosine has been used as a marker to measure extent of DNA oxidative damage. Furthermore, alteration or overexpression of the p53 gene was considered an indication of apoptosis. This article reviews some of the aspects of biomarkers of apoptosis, indicating relevance of their uses to predict apoptosis following exposure to environmental toxicants. PMID- 11503420 TI - Decompression prosthesis. PMID- 11503421 TI - Management of the dental patient with pulmonary tuberculosis. AB - Tuberculosis is a potentially fatal infectious disease. Although it typically manifests in populations subjected to crowding and living in poor conditions of hygiene, in the past decade the morbidity and mortality associated to pulmonary tuberculosis has increased throughout the world--particularly due to the appearance of strains resistant to a range of drugs, and to the vulnerability of human immunodeficiency virus (HIV)-positive individuals to tuberculosis infection. In this context, dental professionals should implement schemes and protocols designed to identify patients with lung tuberculosis, in order to both refer such patients to specialized medical care and to avoid infection of the dental personnel and other patients. A review is made of the etiology, pathogenesis, clinical manifestations, diagnosis and treatment of patients with pulmonary tuberculosis, with particular emphasis on the management measures indicated in the dental clinic. PMID- 11503422 TI - [We attempt to elucidate whether candidosis or candidiasis should be used when referring to disease caused by fungi belonging in the genus Candida]. PMID- 11503423 TI - [Head and neck radiated patients]. PMID- 11503424 TI - Spirituality, meaning, mental health and nursing. PMID- 11503426 TI - Children speak: nursing is "a girl's job". PMID- 11503425 TI - Results of VNS long-term depression study are positive. PMID- 11503427 TI - What every nurse needs to know about nocturnal sleep-related eating disorder. AB - This article provides information about a bizarre pattern of eating while asleep called nocturnal sleep-related eating disorder. People with this disorder, which has begun to be studied only recently, demonstrate features of both a sleep disorder and an eating disorder. Many clients are reluctant to initiate discussions regarding this condition because of feelings of powerlessness and shame. Other clients do not discuss their symptoms because nurses and other clinicians fail to gather accurate assessment data due to lack of knowledge within the professional community regarding the disorder. This review includes the historical background, definition of terms, and clinical manifestations of nocturnal sleep-related eating. In addition, emphasis on assessment and clinical management are included. Safety issues, the need to educate health care providers, and the role of the nurse in advocating for appropriate diagnosis, treatment, and referral are addressed. PMID- 11503428 TI - Revisiting the identity issue in anorexia. AB - Anorexia nervosa is considered an enigmatic disease with a multitude of predisposing factors and no empirically confirmed effective treatment. However, all people with the disease relentlessly pursue a common idealized goal--the state of severe emaciation. In this article, the perspective that this condition epitomizes a life-time struggle for self-acceptance and self-definition is reexamined. People afflicted with anorexia come to define themselves through their emaciated form, and maintenance of this form becomes their major focus and primary means of fulfillment. Therapy must address the identity issue, which is fundamental to the onset and perpetuation of the disease. Use of interpersonal techniques is advocated as most appropriate in serving this purpose. PMID- 11503429 TI - Lithium treatment present and future. AB - 1. Lithium remains unmatched for long-term effectiveness against the morbidity and mortality associated with bipolar I and II disorders. 2. Lithium is the first choice for bipolar depression. 3. Recent studies suggest that long-term lithium treatment may have mortality-lowering and anti-suicidal effects. 4. Chronic lithium, or valproate, administration has been shown to have a neuroprotective effect on rat and human cultured neurons. PMID- 11503430 TI - From safety contract to safety agreement. AB - 1. Confusion and controversy continue to surround the definition and use of safety contracts. 2. A safety agreement based on Orlando's Nursing Theory may be a more practical, concise, and nurse-patient friendly means to promote patient safety. 3. The key ingredients in a safety agreement are identifying patients' immediate safety needs and decreasing patients' immediate distress related to safety through the nurse-patient relationship. 4. The value of a safety agreement lies not in the tool itself but in its promotion of a nurse-patient relationship that involves communication and collaboration. PMID- 11503431 TI - A life journey with schizophrenia. PMID- 11503432 TI - Behind the scenes. PMID- 11503433 TI - [Sp1 transcription factor]. PMID- 11503434 TI - [Dinucleoside polyphosphates: occurrence, metabolism and function]. PMID- 11503435 TI - [Human genome 2001]. PMID- 11503436 TI - [Structure of the prokaryotic ribosome]. PMID- 11503437 TI - [CCC DNA--intermediate replication form of HBV genome]. PMID- 11503438 TI - [The role of GreA and GreB factors in bacterial transcription elongation]. PMID- 11503439 TI - [DNA repair in mammalian cells: mechanisms of homologous recombination]. PMID- 11503440 TI - [Are mitochondrial mechanisms of DNA repair effective?]. PMID- 11503441 TI - [Role of Rsp5p ubiquitin-protein ligase in endocytosis of plasma membrane proteins in yeast]. PMID- 11503442 TI - [Estrogen receptor beta--structure, regulation and function]. PMID- 11503443 TI - [Role of chemokine receptors in HIV infection and AIDS therapy]. PMID- 11503444 TI - [Genomic imprinting in Prader-Willi syndrome]. PMID- 11503445 TI - [Etiology of Angelman syndrome--new views on the mechanism of genomic imprinting]. PMID- 11503446 TI - Telemedicine using standard Internet technologies (telemedicine for less than $1,000). PMID- 11503447 TI - Choosing your telemedicine system. PMID- 11503448 TI - How to make telemedicine work. PMID- 11503449 TI - Teleradiology. PMID- 11503450 TI - Telesonography. PMID- 11503451 TI - Telecytology. PMID- 11503452 TI - Other telemedicine applications. PMID- 11503453 TI - Regulations and other issues. PMID- 11503454 TI - What's ahead? PMID- 11503455 TI - Veterinary telemedicine. Introduction. PMID- 11503456 TI - Why telemedicine? PMID- 11503457 TI - Understanding image-capture devices. PMID- 11503458 TI - [Development of perforator vein surgery from the Linton and Cockett procedure to endoscopic dissection]. AB - Interruption of incompetent perforator veins (PV) is regarded as one of the principles of therapy in cases of severe stages of chronic venous insufficiency. This paper is to demonstrate the development of PV surgery from the original procedure (termed the radical Linton's operation after its originator) to minimal invasive approaches as the subfascial endoscopic perforator vein surgery (SEPS). Subfascial or epifascial interruption is frequently complicated by poor postoperative wound healing (sometimes more than 50%) even in cases with smaller incisions as the former Linton's procedure. PV interruption by hooks and special knives has been in wide use, however, the procedure is not as precise as a direct subfascial approach. The blind subfascial perforator interruption and fasciotomy by Hach has been advocated in severe forms of chronic venous insufficiency. It was Hauer at the beginning of the eighties who introduced a video-controlled surgical approach using a scope with fiberoptic lighting and video camera. At this time, a variety of instruments and modified techniques exists, but the effect of SEPS on venous hemodynamics and clinical outcome is not totally proven due to an additional reflux or obstruction in the deep or saphenous veins and concurrent epifascial surgery. A lower rate of complications of the endoscopic procedures is described, however, there is still a rate of about 5% hematoma and about 7% delayed wound healing. SEPS is an important adjunct in severe cases of PV insufficiency with chronic venous ulcers and lipodermatosclerosis. SEPS will fail in circular crural ulceration where the lesion should be totally excised and covered with skin transplantation. Up to now there is no study to demonstrate the efficacy of SEPS in patients with primary saphenous vein incompetence without deep reflux. PMID- 11503459 TI - [Technique of endoscopic subfascial CO2 insufflation-dissection of perforator veins]. AB - Endoscopic subfascial dissection of perforating veins (ESDP) represents a new additional technique for treatment of greater saphenous system varicosities. We present a modification of ESDP, entitled endoscopic subfascial insufflation dissection of perforating veins (ESIDP). In ESIDP CO2-insufflation induced enlargement of subfascial space (subfascial pneumocave) facilitates identification and dissection of perforating veins. From 1.1.1998 to 1.7.2000, 150 procedures of ESIDP have been conducted in 127 patients (90 females, 37 males). Identification of incompetent perforating and superficial veins and assessment of deep veins were performed preoperatively using duplex sonography. In addition to a description of the surgical technique, we present an overview about indications for ESIDP and possible ESIDP-related postoperative complications. PMID- 11503460 TI - [Varicose vein operation under inpatient conditions]. AB - The change of surgical operations from inpatient to outpatient conditions as a method of lowering costs in health care has lead to a significant rise in the number of outpatient operations, especially in varicose vein surgery. Hospitalisation is still required in special cases. The criteria of 146 patients with varicose veins for admittance to inpatient treatment were retrospectively reviewed. The clinical findings, the designated operation and the specific personal factors were assessed. We found in just under 70% of the patients the indication for hospital admittance whereas in slightly over 30% the outpatient management was ascertained. Through this selection the total number of inpatients could be reduced, leading on the other hand to a relative increase in the number of difficult cases with more severe conditions requiring more complex management. PMID- 11503461 TI - [Ambulatory operation of varicose veins]. AB - Over the past years out-patient operations of varicose veins have become increasingly accepted. In the year 1998 the number of patients treated for long saphenous vein varicosis on an outpatient-basis has risen to 66,922. For the individual risk assessment of the patient the ASA-classification is proven. ASA class I and II patients may be considered for out-patient treatment. 98.2% of our patients that were operated upon 1995 on an outpatient-basis were classified as ASA I or II. Beside the risk assessment the extension of the disease should determine the indication for outpatient or inpatient treatment. During the year 1995 the majority of operations were performed for long or short saphenous vein varicosis (51.3% in the outpatient group, 61.9% in the inpatient group). But the percentage of isolated treatment of sidebranches and perforating veins was much higher in the outpatient group with 30% in comparison to the inpatient group with 3.4%. Under these conditions outpatient varicose vein surgery can be carried out with good result and a low complication-rate. In our outpatient group a deep vein thrombosis occurred in 0.03%, in other outpatient centers the incidence of deep vein thrombosis was reported with 0.08%. Wound infection was seen in 0.49%, a bleeding complication in 0.16% of the patients. PMID- 11503462 TI - [Is there justification for varicose vein surgery with local anesthesia in successive procedures? Prospective study of 5,000 crossectomy/stripping operations]. AB - In a prospective study we examined the complications of 5,000 operations (n = 876 as outpatient; ligations of the SFJ/SPJ +/- stripping or recurrence operation of the SFJ/SPJ), that were carried out under local anaesthesia in a successive procedure. We registered only the in-hospital complications or those occurring during the first postoperative week of outpatient procedure: Mortality, deep venous thrombosis, pulmonary embolism, major vascular injuries, major nerval injuries, anaesthesia complications. The proof and/or exclusion of complications were done by clinical investigations with the parameters: Mortality, vascular and/or nerval injuries, anaesthesia complications as well as pulmonary embolism. With the parameter deep venous thrombosis n = 2,495 operations were examined by color duplex scan in the region of the sapheno-femoral (popliteal) junction before patients were discharged from hospital/ambulatory therapy. In the other patients the diagnosis was done according to clinical parameters. RESULTS: In the investigation period two complications occurred at the nerval-system after difficult ligation of the SPJ: a) Paresis concerning mainly the peroneal branch of the N. ischiadicus with regeneration after nine months. b) Lesion of the N. tibialis with paresis of the M. abductor digiti minimi that was unchanged after seven months. CONCLUSION: Varicose vein surgery under local anaesthesia as a successive procedure represents a very safe treatment option. A comparison with literature data from prospective or retrospective studies is not possible, because of missing studies and/or unclear information or definition problems. It would be desirable if such studies could be performed. PMID- 11503463 TI - [Tumescence local anesthesia in venous surgery]. AB - After the long standing application of the tumescent solution with percutaneous sticks the tumescent technique in combination with the Rofil Medro-pump represents at the moment a very elegant and innovative method of local anesthesia in the field of phlebosurgery. Since 1997 the tumescent technique has been used in our hospital for all major surgeries in the field of phlebosurgery. Even more difficult operations like inguinal relapse-varicosis or acute thrombophlebitis are successfully performed with this method. From our experience the advantages of this method are a reduced bleeding, less hematomas, an insignificant risk of thrombosis and embolism, an antibacterial effect, the hydrodissection and anodynia. Using very large volumes painless surgical treatment of complicated and extended findings is possible. Essential disadvantages (like a wet surgical field, which needs to get used to) hardly exist neither for the patient nor for the surgeon. Therefore, the tumescent technique represents a safe, comfortable and almost painless kind of local anesthesia of the skin and the subcutaneous fatty tissue. PMID- 11503464 TI - [Recurrent varicose veins. Surgical procedure--results]. AB - We report 96 patients (107 legs) with recurrence after varicose vein surgery. We define a recurrent varicose vein as a new transfascial insufficiency after incomplete interruption of the saphenofemoral as well as popliteal junction with reflux in the epifascial venous system. 92 patients underwent external primary surgery. Morphologically an insufficient high ligation of the V. saphena magna (Moszkowicz's operation) led in 30 cases to a recurrent varicose vein. In 47 cases an incomplete isolated, in 29 cases an incomplete saphenofemoral ligation during a Babcock procedure and in one case an incomplete saphenopopliteal ligation for V. saphena parva insufficiency were the reasons for the recurrence. As recurrent surgery we performed 106 ligations of the saphenofemoral junction. In 38 of these cases an isolated saphenofemoral ligation and in 68 cases an additional stage-adjusted ligation of the V. saphena magna were carried out. In one case of recurrent saphenopopliteal insufficiency a repeated ligation of the V. saphena parva was performed. The recurrence is an avoidable complication of a not perfect primary surgery. The main cause is an inadequate access with incomplete saphenofemoral or -popliteal ligation. PMID- 11503465 TI - [Surgery of the lateral accessory saphenous vein]. AB - Confusion exists regarding the right surgical strategy for therapy of isolated varicosis of the lateral accessorian saphenous vein (LASV) resulting from an incompetent valve at the crosse without concomitant reflux into the long saphenous vein (LSV). In a retrospective study 30 patients with isolated varicosis of the LASV were examined 3 years after surgical treatment. In this study the first segment of the LSV was removed after resection of the LASV. 64% of our patients were free of recurrence after 3 years. 25% had partial reflux from the Boyd perforans vein and only 9% of the patients had a complete reflux in the LSV. Based on our results it is justified not to remove the LSV in cases of isolated varicosis of the LASV. PMID- 11503466 TI - [Prospective study of avoiding neoangiogenesis after great saphenous vein crossectomy. Initial results]. AB - A meticulous dissection of the sapheno-femoral junction (SFJ) at the time of primary surgery is regarded as best protection against the development of recurrences from this area. However, despite correct ligation of the junction recurrences may occur. In a prospective randomised trial, which has been started in 1998, we want to find out, whether this regrowth might be inhibited by the use of different ligation technics of the SFJ: Group 1: Ligation of the SFJ with resorbable Vicryl Group 2: Ligation with Vicryl and continuous non-resorbable stitching over (Prolene) the saphenous stump which precludes contact between free stump endothelium and the surrounding subcutaneous tissue Group 3: Non-resorbable ligation of the SFJ (Ethibond) Group 4: Ethibond ligation with Prolene stitching over the saphenous stump. The first follow-up examinations, which have been done by color-dupley-scan are presented. In each group about n = 100 groins could be examined. We found slight inguinal insufficiency due to a small branch of the femoral vein in group one: n = 10; group two: n = 6; group three: n = 3; group four: n = 1. Our hypothesis that the use of the suture material or the free lying stump endothelium might be influencing the development of the neovascularisation, seems to be supported by these results. We found the lowest rate of postoperative inguinal refluxes in the Ethibond-Prolene group, where the contact between free stump endothelium and the surrounding subcutaneous tissue is precluded. However, definite reliable data will be presented not before the third follow-up (24 months after the operation). PMID- 11503467 TI - [Stage-adapted therapy concept in ascending thrombophlebitis]. AB - Varicophlebitis is the most frequent and important acute complication of a varicosed long and/or short saphenous vein. In view of the controversial discussion about the either conservative or surgical treatment, a clinically relevant classification of this syndrome appears useful: Stage I includes varicophlebitis without involvement of the respective junctional valve--in the groin or at the knee--and deep veins. While in Stage II the proximal part of the thrombus has reached the respective junctional valves of the long or short sapheneous vein, in Stage III it has entered the deep veins by means of these valves. In Stage IV the thrombus migrates via insufficient perforating veins into the deep system. Stages I and IV should be treated conservatively first, removal of the varicous veins should be performed after regression of the acute symptoms. Stages II and III should be considered an indication for urgent surgery. The surgical strategy consists of crossectomy, resection of the saphenous vein without stripping, radical excision of all varicous veins and ligature of insufficient perforating veins. In stage III the thrombectomy of the deep veins using the Fogarty-procedure must be carried out before any other measures are taken. In 1996 a total number of 40 limbs with ascending varicophlebitis (stage I = 16, stage II = 19, stage III = 5) was observed. 10 extremities (stage I = 2, stage II = 5, stage III = 3) underwent surgical treatment. 1 patient developed a deep infection of the groin, the average stay in hospital was 9 days. PMID- 11503468 TI - [Complications in varicose vein operations]. AB - Although operations on varicous veins are regarded as easy and low-risk, severe complications with the involvement of arteries, veins and nerves can occur during the course of such operations and, in reconstruction, demand the whole spectrum of vascular surgery. Therefore, surgeons operating on varicous veins and on out patients should possess sufficient experience in vascular surgery and be able to admit their patients to a centre of vascular surgery in the case of severe complications without unnecessary loss of time. Beside a perfectly done operation sufficient preoperative diagnostic examinations and postoperative care of the patient have to be claimed. The treatment of varicous veins in a hospital must be possible also in the future. The extent of the procedure is usually greater during stationary treatment and bilateral operations are feasible. Therefore, from the economic point of view, a stationary assignment for a few days does not seem more expensive than several single operations with subsequent outpatient treatment and repeated diagnostic examinations in ambulatory settings. PMID- 11503469 TI - [Complications in surgery of varicose veins]. AB - Based on our experience derived from approximately 20,000 varicose vein operations during the last three decades we report about possible complications in varicose vein surgery. We had no postoperative mortality. Crossectomy of the greater saphenous vein may lead to major complications. We describe a case of femoral artery injury. Postoperative deep vein thrombosis and pulmonary embolism are very rare events. Bleeding complications in the groin, necessitating surgical reintervention are seldom, other bleeding complications like suffusions and hematomas can be seen more often. These complications are significantly more frequent if we use low molecular weight heparins for prophylaxis of deep vein thrombosis postoperatively. Lymphatic complications like lymphcysts and -fistulas are harmless in most cases and regress spontaneously. Concerning neurological complications lesions of the saphenous nerve are of some importance. Altogether the risk for a patient undergoing varicose vein surgery is minimal but not zero. PMID- 11503470 TI - [Indications and technique for sclerotherapy of varicose veins]. AB - Sclerotherapy is in general a very safe method for the treatment of teleangiectasias and reticular varicose veins done by an experienced therapist. The complication-rate is very low, with the use of Polidocanol the rate of skin necrosis ranges between 0.001 and 0.2%, the incidence of superficial thrombophlebitis is 0.08%, the incidence of an allergic reaction is reported with 0.2%. The sclerotherapy of large perforating veins and the long or short saphenous veins is controversially discussed. The results of prospective randomized studies show better results with operative treatment in comparison to sclerotherapy. The recurrence-rate of sclerotherapy of the long saphenous vein varies between 22 and 37% after 3 years, between 40 and 70% after 5 years and 94% after 10 years. Before starting the sclerotherapy the extension of the varicose vein disease should be diagnosed correctly. The diagnosis can be obtained with duplex-ultrasound, that gives either functional and morphologic information about the deep and superficial venous system. PMID- 11503471 TI - [Medical compression therapy]. AB - In order to evaluate the correlation between the pressure exerted by compression stockings and a resulting improvement in venous hemodynamics, we carried out an open, randomized, prospective study on 22 patients (11 women and 11 men with an average age of 55.1 [10.3]) with chronic venous insufficiency (CVI) in the clinical stages C1-4, Ep, AS, Ap, PR. Dynamic strain gauge plethysmography was used to measure the acute effect on venous hemodynamics of 9 different compression stockings in compression class 2 (A-D). At the same time when venous function parameters were monitored, we also measured the pressure exerted by the compression stockings during rest and exercise. With all compression stockings the average resting pressure fulfilled in reclined patients the specifications for compression class 2 (25-35 mmHg at the ankle, CEN). The compression stockings lengthened venous refill time t0 in a statistically significant degree. The improvement in venous function was correlated with the ratios of maximal working pressure to resting pressure while standing (r = 0.97, p < 0.001). Compression stockings belonging to the same compression class vary in their acute effect on venous hemodynamics. The efficiency of the different therapeutic compression stockings was largely dependent on the amount of fabric stretch, which can be characterized in vivo with the ratio of maximum exerted pressure during movement to that while standing still. A knowledge of the hemodynamic effectivity of the various compression stockings allows the optimal stocking selection for each patient and his individual clinical situation. PMID- 11503472 TI - [Comments on the history of surgery in Strassburg (I)]. PMID- 11503473 TI - [Treatment of pelvic injuries (1)]. PMID- 11503474 TI - MedBytes. PMID- 11503475 TI - 'On our own terms'--one year later. PMID- 11503476 TI - Managing the pain of advanced chronic disease. AB - Persons with life-limiting illness such as cancer, congestive heart failure, chronic obstructive pulmonary disease, and other organ system failure may needlessly endure severe pain for months or even years as the disease progresses. Adequately assessing and treating pain and other factors associated with advancing disability often dramatically alters the patient's experience of the disease process. Regulations concerning the use of controlled substances for the treatment of chronic pain are reviewed, and a simple yet thorough pain assessment and treatment strategy is presented. Physicians practicing these palliative skills will be gratified when caring for patients with chronic, life-limiting illness, as the potential exists for improving patient well-being, even in the face of advancing disease and ensuing disability. PMID- 11503477 TI - Symptom management in hospice and palliative care. AB - Managing the symptoms of advanced disease at the end of life is one of the most challenging aspects of medicine for most clinicians. Traditional textbooks provide limited resources for treating patients at this stoichiometric point in their disease. This article provides an overview in the treatment of common symptoms at the end of life, such as anxiety, anorexia and cachexia, constipation, delirium, dyspnea, fatigue and asthenia, nausea and vomiting, malignant intestinal obstruction, and terminal restlessness. By addressing these symptoms, the physician can play a key role in the patient's achievement of a peaceful, symptom-free, and dignified death in the setting of their choice. PMID- 11503478 TI - Providing hospice care in rural Texas. AB - Hospice organizations in rural areas face many logistical and financial challenges. The authors review their experiences in offering hospice care to a widely dispersed population of patients. Crown of Texas Hospice, headquartered in Amarillo, Texas, provides hospice care throughout most of the vast and sparsely populated Texas panhandle. In the United States, patients typically are not referred to hospice care until the very end of the terminal phase of their disease process. Late referrals limit the opportunities for optimal symptom management and also limit the ability of a hospice organization to meet the psychosocial and spiritual needs of patients and their families. Recommendations to improve the timeliness of hospice referral are suggested. PMID- 11503479 TI - Spiritual care at the end of life. AB - An essential part of end-of-life medical care, spiritual care allows physicians to recognize problems like meaninglessness, anguish, and hopelessness, for which effective interventions are available. This form of care can be a deeply rewarding area of medical practice. PMID- 11503480 TI - New programs for children living with life-threatening conditions. AB - Palliative care is not commonly available to most children who die. Some children who die need comprehensive child- and family-centered services for all their lives, or for several years, followed by bereavement care. Other children who die have needs that are emergent, acute, and short term with regard to medical care but very long term with regard to bereavement needs. Exciting new initiatives may soon change the experience of children living with life-threatening conditions and their families. PMID- 11503482 TI - A nursing home visit. PMID- 11503481 TI - Research opportunities in palliative medicine. AB - Patients who require palliative care have many physical and psychosocial needs. Their families also require care and advice. Despite recent advances in assessment and management, major issues relating to palliative care for patients and their families remain unanswered. Several challenges face researchers in this area. We discuss administrative issues, including historical development, funding, academic recognition, integration of services, and cooperation among different centers. Methodological challenges in patient issues and trial design are outlined, and areas for future research are proposed. PMID- 11503483 TI - [Should angioplasty still be considered for atherosclerotic renal artery stenosis?]. PMID- 11503484 TI - [The peony, a forgotten panacea]. PMID- 11503485 TI - [Anatomy and basic knowledge of the spinal cord]. AB - Basic knowledge of macroscopic anatomy is needed to understand the clinical signs and distinct syndromes resulting from the disturbed function of the tracks, segments and roots of the spinal cord. The cord is contained within the vertebral canal, continuing the medulla down to the conus medullaris. Descending tracks are motor (corticospinal and extrapyramidal), and ascending tracks are sensory (spinothalamic and lemniscal). Spinal cord involvement must be considered when a sensorimotor spinal tract and a radicular spinal syndromes are combined. Syringomyelia, Brown-Sequard syndrome and subacute combined degeneration of the spinal cord must also be familiar to clinicians, as well as how to differentiate a conus medullaris from a cauda equina syndrome. PMID- 11503486 TI - [Mechanical pathology of the spinal cord]. AB - Syringomyelia is a cyst within the spinal cord, associated with numerous causes, most frequently combined with the Chiari abnormality. Many other causes of craniovertebral junction anomalies are described. In those cases the syringomyelia is so-called hind-brain related syringomyelia. Other dysraphic states and spinal causes of syringomyelia are spina bifida cystica, lumbar lipomas, diastematomyelia or split notochord syndrome. Mechanical causes of spinal cord compression are mostly of tumoral origin. The vast majority of spinal metastases occur extradurally and originate from carcinomas of the breast, prostate and lung. Intradural extramedullary tumours of the spine constitute two thirds of all spinal neoplasm: schwannomas and meningiomas make up approximately 90% of the total and occur in equal numbers. PMID- 11503487 TI - [Inflammatory pathology of the spinal cord]. AB - Diagnosis of myelopathy whatever its cause, is based on clinical findings: paraparesis, loss of sensibility, sphincterian disturbance. Diagnosis of myelopathy requires magnetic resonance imaging, which rules out compressive and vascular disorders. Main causes of inflammatory myelopathies are multiple sclerosis multisystemic disorders, infectious and post-infectious diseases. Absence of identifiable causes in one quarter of the cases leads to the diagnosis of idiopathic myelitis. PMID- 11503488 TI - [Pathologic vascular malformations of the spinal cord]. AB - Like for the intracranial level, spinal cord vascular malformations can be classified, according to their radiological (particularly arteriographic) and histological aspects, as: arteriovenous malformations, cavernomas and so-called malformations, arterial aneurysms, and venous malformations. This classification should also include the involvement of the surrounding envelopes. Clinical manifestations are not always typical. Magnetic resonance imaging is useful for diagnosis, and angiography is not always necessary nowadays. Arteriovenous malformations are the most frequent vascular malformations; they are equally represented by arteriovenous fistulas of the dura mater and intradural arteriovenous malformations. Intradural cavernomas are rare as compared to arteriovenous malformations. Intradural arterial aneurysms are a rare, owing to local haemodynamic conditions. Venous malformations are exceptional; some intramedullar forms have been recently reported. PMID- 11503489 TI - [Neurodegenerative diseases of the spinal cord]. AB - Numerous classifications have been applied to describe degenerative diseases of the spinal cord before the era of molecular analysis. The present challenge is to match in an appropriate manner old and new clinical classifications with the results of genetic analysis. This work is now well advanced for spinocerebellar degeneration but needs to be developed for hereditary spastic paraplegia, spinal muscular atrophies and amyotrophic lateral sclerosis. The widening of clinical spectrum observed in hereditary neurodegenerative diseases of the spinal cord raises the question of modulating factors (environmental or genetic). Clinicians in charge of patients with progressive spinal cord signs should bear in mind that the field of degenerative diseases is a moving one and that a degenerative aetiology has to be suspected more widely than in the past. PMID- 11503490 TI - [Spinal cord infections]. AB - Although infectious myelopathies are rare, appropriate aetiological diagnosis is of crucial importance to improve outcome. Lyme disease causes a myelomeningoradiculitis during the second stages and a progressive encephalomyelitis during the third stage. Serum and cerebrospinal fluid antibody tests are the basis of diagnosis. Myelopathy usually responds to treatment with ceftriaxone or doxycycline. Vacuole myelopathy occurs in patients with human immunodeficiency virus infection. It is characterised by a progressive spastic and ataxic paraparesis. About half of patients have symptoms of dementia. No specific treatment is available. Human T-cell lymphotropic virus type I is endemic in tropical areas and particularly in French West Indies. It causes a chronic spastic paraparesis with sexual and bladder dysfunction. Diagnosis depends on the positive antibody response in both serum and cerebrospinal fluid, and on the exclusion of other causes of spinal cord disease. There is no specific treatment. PMID- 11503491 TI - [Intramedullary tumors]. AB - Intramedullary tumours represent 2-4% of all central nervous system tumours. Clinical presentation can adopt any aspect of slow medullar compression. Often benign, these tumours are mostly of glial origin: ependymomas or astrocytomas. Diagnosis has been revolutionised by magnetic resonance imaging, which allows a precise analysis of the tumour with differentiation between the consistent portion and its satellite cysts. Surgery remains the treatment of choice. Total removal can often be achieved and no further complementary treatment is justified, except in proven malignancy. If resection has been partial, radiotherapy remains controversial as non-deprived of untoward effects, when delivered at efficient dose. Long term follow-up is mandatory, as there are late recurrences or secondary aggravations, evolving on their own. As post-therapeutic clinical state is highly correlated with the pretherapeutic state, intramedullary tumours are to be considered as medical emergencies. PMID- 11503492 TI - [Pernicious anemia]. AB - Pernicious anaemia is the most common cause of cobalamin deficiency. Nervous disorders associated with cobalamin deficiency are neuropathy, optic atrophy, dementia and myelopathy (subacute combined degeneration). In this case, symptoms are those of posterior and lateral column dysfunction of the spinal cord, with diminished vibratory sensation, ataxia, weakness of limbs, hyperreflexia, extensor plantar response and spasticity. Macrocytosis and anaemia are often lacking. There is an inverse correlation between the degree of anaemia and the extent of nervous impairment. The most sensitive tool for the diagnosis of cobalamin deficiency is the serum cobalamin level. But a normal cobalamin assay does not fully exclude cobalamin deficiency. Levels of serum methylmalonic acid and total homocysteine are useful as ancillary tests in the diagnosis. Treatment is based on intramuscular injections of vitamin B12. PMID- 11503493 TI - [Diffuse interstitial pneumonia. Etiology and diagnosis]. PMID- 11503494 TI - [Acute renal failure. Etiology, physiopathology, diagnosis, principles of treatment]. PMID- 11503495 TI - [Breast cancers. Epidemiology, pathology, staging, diagnosis, etiology, principles of treatment]. PMID- 11503496 TI - [Chronic viral hepatitis B and C. Epidemiology, diagnosis, etiology, prevention]. PMID- 11503497 TI - [Psychiatric disorders in pregnancy and postpartum. Diagnosis]. PMID- 11503498 TI - [Radiotherapy. Radiobiological aspects, principal secondary effects]. PMID- 11503499 TI - [Treatment of scabies]. PMID- 11503500 TI - [Pierre Dionis, a surgeon in Moliere's time]. PMID- 11503501 TI - [The transfusion chain: from donor to recipient]. AB - From donor to recipient, raw blood will undergo a succession of processing events before eventually becoming a qualified, recipient-adapted finished product. The integrity of the transfusion chain and its various links will determine the final quality of blood transfusion, a key element for a number of medical disciplines. The recent reform of blood transfusion has brought about a national system based on a single body, the French Blood Institution, with regional branches ensuring effective networking of the entire country. Keeping a permanent scientific watch, this Donor-to-Recipient set-up warrants optimal quality according to current knowledge, and close links with the single hemovigilance network which continually analyses events that may affect transfusion safety and takes immediate action accordingly. PMID- 11503502 TI - [Clinical and biological qualifications for blood donations]. AB - Three major steps are very important to increase transfusion safety: epidemiological step: donor selection; biological step: qualification of each blood donation; technological step: obtention of final product. The final therapeutical blood products must meet the requirements concerning their quality and safety, two factors with which the population is very demanding. These requirements can be ensured only if ethical, clinical epidemiological and biological parameters are clearly defined. The strict application of these criteria to each step of the transfusion process, from the blood donation to the final product, allows us to ensure the security of recipients with regard to known risks but also to anticipate other emergent risks. PMID- 11503503 TI - [Labile blood products and their preparation]. AB - Licensed labile blood components are put down on a regulatory list by the French Health Authority. They are prepared by the French National Blood Service and controlled according to regulatory and validated procedures. Depending on the origin of the blood, labile blood components are either homologous (donors) or autologous (from patients). Blood components (red cell concentrates, platelet concentrates and fresh frozen plasma) are processed within sterile closed disposable systems using either post-donation processing of whole blood or apheresis technology. All homologous blood components are leuco-reduced. Depending on specific settings, blood components could be washed, gamma irradiated or cryopreserved. Fresh frozen plasma is either "donor retested" after quarantine or viro-attenuated by solvent detergent treatment. Release is only allowed after a full conformity control of each blood component unit. PMID- 11503504 TI - [Immune safety in blood transfusions]. AB - Immune security of blood donation is defined by all means aiming at reducing or eliminating the immune risk related to blood donation. It concerns the whole transfusion process from the blood donor to the receiver. The immune risk of blood donation is directly related to the polymorphism of molecular and cellular blood groups systems. Immune security consists in avoiding the meeting of antigens and augmented by the direct baneful consequences of the immune conflict. This requires the previous immune characterisation of blood products and of the patients and of their compatibility, which must be strongly maintained along the transfusion process. To control this process, which is still too much deficient, represents the true guarantee of immune security of blood donation. PMID- 11503505 TI - [Indications for labile blood products]. AB - The aim of blood transfusion therapy is to supply specific labile products to patients in quality and in quantity. Different basic blood products are available: red blood cell concentrates, platelets concentrates and fresh frozen plasma, and according to their biological parameters, some of them can be selected and secondly adapted. Therefore, because some blood products are very rare and expensive, it is not possible to systematically use blood products having all the specifications. Also, it is necessary for clinicians to know blood product indications in order to avoid inadapted and abusive prescriptions inducing blood product unavailability damaging for other patients. The benefice risk ratio must be continuously measured, because these human products have some adverse effects: specially immunologic reactions and transfusion transmitted diseases. The systematic cell blood product leukoreduction since April 1st 1998 in France, allowed us to decrease these risks and to obtain a good tolerance. The hypothetical prion blood product transmission must be taken into consideration by plasma leukoreduction and limiting the blood product transfused number, in the respect of each specific clinical situation. PMID- 11503506 TI - [Autologous blood transfusion]. AB - Autologous blood transfusion techniques are the principal means of reducing allogeneic blood exposure. Those techniques were developed in order to prevent the risk of contamination by viruses, mainly HVB, HCV and HIV. However that risk has become so small that all studies show an exorbitant cost/efficiency ratio. Autologous blood transfusion would therefore be of no interest in terms of public health but a recent experimental study suggested a possible transmission of the BSE agent through blood. Until the matter is settled, the precaution principle means we should prefer alternative techniques to allogeneic blood whenever possible, hence a renewed interest in autologous transfusion. PMID- 11503507 TI - [Blood transfusion surveillance: organization and results]. AB - Created by law in 1994, haemovigilance is a national system of surveillance and alarm, from blood collection to the follow up of the recipients, gathering and analysing all untoward effects of blood transfusion in order to correct their cause and prevent recurrence. Three levels compose the haemovigilance network: local, regional and national. The incident reporting is mandatory and this rate is stable with 2.9 to 3 incidents per 1000 blood components transfused. The transfusion incident due to bacterial contamination and ABO incompatibility worry the medical community. Haemovigilance contributes to transfusion safety. It makes possible to recognise risks already known, to be on alert for emergent risks, to undertake preventive actions on critical steps of the transfusion chain, and to warrant follow up of safety measures efficiency. PMID- 11503509 TI - [The researcher, the clinician, the teacher ... and the evaluation]. PMID- 11503508 TI - [Challenges of transfusion medicine]. AB - Transfusion medicine has the logic of a therapeutic chain applied to labile blood components and cell therapy products, within a coherent structure, such as the recently created Etablissement francais du sang. Faced to the threat of emerging- sometimes hypothetical--transfusion risks, such as the possible transmission of BSE by blood transfusion, the precaution principle requires developing strategies to reduce labile blood components consumption, by strictly defining the framework of blood transfusion prescription and encouraging the search for red blood cell and platelet substitutes. In the field of alternatives to labile blood components, research has however yielded few results. The future of transfusion medicine lies in biotechnology: cell (and gene) therapy will become part of novel therapeutic strategies for the treatment of numerous pathologies in man. Transfusion medicine will have to consider the significant advances achieved over the last few years in the field of multipotent stem cells. Transfusion medicine will thus find its place in the promising field of innovating therapies. PMID- 11503510 TI - [Facial traumas. Diagnosis of lesions, early complications]. PMID- 11503511 TI - [Bacterial infections of the newborn. Diagnosis, principles of treatment, prevention]. PMID- 11503512 TI - [Feeling faint, syncope and brief loss of consciousness. Diagnostic approach]. PMID- 11503513 TI - [Hyperlipoproteinemias. Epidemiology, etiology, physiopathology, diagnosis, treatment]. PMID- 11503514 TI - A simple method for the purification of an antimicrobial peptide in recombinant Escherichia coli. AB - A magainin derivative, designated MSI-344, was produced in Escherichia coli as fusion protein, by utilizing a truncated amidophsphoribosyltransferase of E. coli as a fusion partner. Bacterial cells transformed with the gene encoding the fusion protein were grown to a high cell density and induced with isopropyl-1 thio-b-D-galatoside (IPTG) to initiate product expression. The fusion protein was accumulated into cytoplasmic inclusion body and recombinant MSI-344 was released from the fusion partner by hydroxylamine treatment. Following cleavage of the fusion protein with hydroxylamine, the released MSI-344 was purified to homogeneity by cationic exchange chromatography. The final purity was at least 95% by reversed-phase high performance liquid chromatography (RP-HPLC). Purified recombinant MSI-344 was found to be indistinguishable from the synthetic peptide determined by amino acid sequences and antimicrobial activity assay. PMID- 11503515 TI - EBI databases and services. AB - The EMBL Outstation-European Bioinformatics Institute (EBI) is a center for research and services in bioinformatics. It serves researchers in molecular biology, genetics, medicine, and agriculture from academia, and the agricultural, biotechnology, chemical, and pharmaceutical industries. The Institute manages and makes available databases of biological data including nucleic acid, protein sequences, and macromolecular structures. It provides to this community bioinformatics services relevant to molecular biology free of charge over the Internet. Some of these databases and services are described in this review. PMID- 11503517 TI - Flexible genetic engineering using RecA protein. AB - With the explosion in genetic information and almost complete sequencing of the human genome, a shift in the experimental goals of molecular biologists is occurring. Instead of focusing on single genes, current attempts seek to divine the interactions of several genes and sequences. This requires increasingly complex genetic constructs and manipulations, often of very large DNA constructs, and these can be made with RecA protein-based techniques. When RecA protein combined with an oligonucleotide acts as a sequence-specific "masking tape" to block DNA from the action of DNA modifying enzymes, and can be used to direct the cleavage of DNA at single predetermined restriction endonuclease sites. This reaction is called RecA-Assisted Restriction Endonuclease (RARE) Cleavage. The reverse reaction, known as RecA-Assisted Ligation, can be used to join any two desired fragments. When one of those fragments is a vector, a desired fragment can be cloned directly without constructing a genomic library. The reagents and equipment needed are relatively inexpensive, and almost any desired genetic construct up to about 300 kb in size can be made in a straightforward manner. PMID- 11503518 TI - Transcriptional targeted gene therapy for hepatocellular carcinoma by adenovirus vector. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies with poor prognosis and is highly amenable to the development of novel therapeutic strategy. The human alpha-fetoprotein (AFP) gene is normally expressed in fetal liver and is transcriptionally silent in adult liver but overexpressed in HCC. In order to destroy AFP-producing HCC specifically, replication defective adenoviral vectors containing the transcriptional control elements of the AFP gene were designed. Expression of suicide genes by the AFP promoter/enhancer induced prodrug sensitivity in AFP (+) cells but not AFP (-) cells. The expression of suicide genes by ubiquitous promoter, however, showed no selectivity after prodrug treatment. Adenoviral vector transduced genes efficiently not only in vitro but also in vivo, and AFP-producing HCC xenografts regressed by transduction with transcriptionally targeted vectors and subsequent systemic administration of prodrug in animal model. Utilization of the transcriptional regulatory element to drive drug sensitive genes can be a promising strategy for cancer specific therapy. PMID- 11503516 TI - Pathophysiological mechanisms of asthma. Application of cell and molecular biology techniques. AB - Asthma is a common increasing and relapsing disease that is associated with genetic and environmental factors such as respiratory viruses and allergens. It causes significant morbidity and mortality. The changes occurring in the airways consist of a chronic eosinophilic and lymphocytic inflammation, together with epithelial and structural remodeling and proliferation, and altered matrix proteins, which underlie airway wall narrowing and bronchial hyperresponsiveness (BHR). Several inflammatory mediators released from inflammatory cells such as histamine and cysteinyl-leukotrienes induce bronchoconstriction, mucus production, plasma exudation, and BHR. Increased expression of T-helper 2 (Th2) derived cytokines such as interleukin-4 and 5 (IL-4, 5) have been observed in the airway mucosa, and these may cause IgE production and terminal differentiation of eosinophils. Chemoattractant cytokines (chemokines) such as eotaxin may be responsible for the chemoattraction of eosinophils to the airways. The initiating events are unclear but may be genetically determined and may be linked to the development of a Th2-skewed allergen-specific immunological memory. The use of molecular biology techniques on tissues obtained from asthmatics is increasing our understanding of the pathophysiology of asthma. With the application of functional genomics and the ability to transfer or delete genes, important pathways underlying the cause if asthma will be unraveled. The important outcome of this is that new preventive and curative treatments may ensue. PMID- 11503521 TI - University of Pittsburgh and Cephoid develop rapid genetic test that may help cancer patients avoid second surgeries. PMID- 11503520 TI - High level of expression of the Toxoplasma gondii-recombinant Rop2 protein in Escherichia coli as a soluble form for optimal use in diagnosis. AB - The rhoptry 2 protein (Rop2) is an interesting protein of Toxoplasma gondii that is involved in the parasite invasion of host cell, it has three T-cell epitopes and high antigenic value. However, the expression of Rop2 as a recombinant protein in Escherichia coli is not an easy task, showing low levels of expression or degradation and solubility problems. Using a recombinant Rop2(196-561) fused to 6 histidine residues, we showed high levels of expression in bacteria growing in terrific broth. rRop2(196-561) was purified mainly as a soluble product and in high concentrations (approx 1 mg/mL) under native conditions (40 mM imidazol in phosphate buffer). However, after a cycle of freezing-thawing rRop2(196-561) became insoluble. When glycerol was added to 26%, immediately after purification, the protein stayed soluble after cycles of freezing-thawing. Finally, it was demonstrated that under these conditions soluble rRop2(196-561) keeps its diagnostic value in contrast with the insoluble protein. PMID- 11503519 TI - Chemical construction of immunotoxins. AB - Immunotoxins are chimeric proteins consisting of an antibody linked to a toxin. The antibodies most frequently used for the preparation of immunotoxins are murine monoclonal antibodies belonging to IgG isotype. The most used toxins for the chemical construction of immunotoxins are Ricin toxin A chain in its deglycosylated form and recombinant Pseudomonas endotoxin with the cell-binding domain deleted. The linkage of the antibody to the toxin can be accomplished by chemical methods using reagents that crosslink antibody to toxin. The usual crosslinkers attach disulfide groups into the antibody molecule to form a disulfide bond between the antibody and the toxin. Disulfide bonds are susceptible to reduction in the cytoplasm of the targeted cells thereby releasing the toxin so that it can exert its cytotoxic activity only into the cells (e.g., tumor cells) binding the antibody moiety. This article describes various methods to obtain antibodies and toxins and several procedures for their crosslinking as well as "in vitro" and "in vivo" testing of the immunotoxins efficacy. PMID- 11503522 TI - Cepheid's DNA analysis system may help stop vine disease threatening California's $2.8 billion wine & grape industry. PMID- 11503523 TI - [Measuring arterial hypertension]. PMID- 11503524 TI - [Pulse pressure and cardiovascular risk]. PMID- 11503525 TI - [Prognostic value of "white coat syndrome" arterial hypertension]. PMID- 11503526 TI - [The kidney and hypertension: cause or victim?]. PMID- 11503527 TI - Hypertension and dyslipidemia. PMID- 11503528 TI - Microalbuminuria in essential hypertension. PMID- 11503529 TI - [Basic principles of treatment of hypertension]. PMID- 11503530 TI - Non-pharmacologic management of hypertension. PMID- 11503531 TI - Benefits of angiotensin converting enzyme inhibition and angiotensin II receptor antagonists in hypertension. AB - It is through blood pressure lowering that reductions in cardiovascular morbidity and mortality are achieved. The treatment of hypertension patients should also focus on prevention and management of cardiovascular complications, not merely on achieving a target level of blood pressure. In making treatment decisions it is appropriate to assess patients for the presence or absence of risk factors and/or clinical cardiovascular disease. Clinicians should strive to use the least intrusive means possible to achieve goal blood pressure and the treatment regiment should be individualized for each patient. The review has illustrated specific clinical conditions where the use of an hypertensive agent that inhibits the renin-angiotensin system may offer added benefit to the patient while producing the needed anti-hypertensive effects. PMID- 11503532 TI - [Arterial hypertension, menopause and hormone substitution]. PMID- 11503533 TI - A practical guideline for management of hypertension in patients with diabetes. PMID- 11503534 TI - Post-transplantation hypertension. PMID- 11503535 TI - Proceedings of the 1st ASEAN Symposium on Problem-Based Learning in the Health Services. 20-22 November 2000, Singapore. PMID- 11503536 TI - Is problem-based learning a quality approach to education in health sciences? AB - The Faculty of Health Sciences at McMaster University has pioneered, experimented and finally excelled in the application of problem-based learning (PBL) as an entire medical curriculum for the past 35 years. However, the general practice of PBL by other medical schools around the globe has progressed slowly. In theory, PBL as an educational philosophy has long been considered as a quality cognitive concept and was adopted by many medical schools via curriculum reform to improve students' learning attitude. In practice, what is the experimental evidence for PBL meeting the expectation of a quality education in health sciences? How do we differentiate problems associated with PBL philosophy per se from those associated with the ways PBL are handled and implemented? I will address these questions from the perspective of the assessment of performance of students, graduates and practising physicians from the PBL track compared to those from the conventional track based on literature information. Ample evidence suggests that PBL is superior in producing more compassionate physicians and graduates with lifelong learning and leadership quality. But, some educators and administrators are still skeptical that the benefits from PBL may be too marginal to justify the resources required in sustaining it. In this presentation, the assessment of PBL, in both theoretical and practical terms, will be discussed using McMaster PBL as a convenient example because of its relatively long history in practising PBL in medical education. PMID- 11503537 TI - Assessment in problem-based learning (PBL). AB - Problem-based learning (PBL), a relatively new approach to teaching, has been introduced in several of the medical schools in the Southeast Pacific region. This approach emphasises active, self-directed learning and puts the emphasis on the individual student and his/her role in the educational process. It is well known that assessment plays a large role in influencing student learning behaviour. Therefore, it is important that the assessment process not hamper learning or adversely affect attainment of the goals of the curriculum. If student behaviours are directed toward achieving success on the assessments, instructors' efforts to create a climate of self-directed learning and individual responsibility will be frustrated. This article provides information on the different classes of assessment vehicles available to assess the various domains of medical competence. We pay special attention to the critical relationship between the educational goals of problem-based learning and the different assessment instruments used. We briefly describe some assessment instruments and how it can relate to these goals in a positive manner. We conclude that no assessment instrument is ideal and that each has unique inherent deficiencies. As a consequence, a successful student assessment process requires use of several complementary instruments. PMID- 11503538 TI - Clinical reasoning learning sessions. AB - Problem-based learning (PBL) is used all over the world and is recognised as a modern and efficient way of teaching medicine. In most medical schools, these techniques are applied during the pre-clinical phase of the curriculum. We report here our experience with PBL during clerkship rotations. These sessions, called Clinical Reasoning Learning (CRL), have, as a main objective, the organisation of knowledge already stored by our students in mental networks, using a well-defined strategy. The principle of these sessions is to recreate a clinical consultation setting focussed on a specific problem identified among the mandatory objectives of rotation. Based on examples taken from the urological content of teaching during the rotation in surgery, we demonstrate the method and discuss the basis of this technique. PMID- 11503539 TI - Problem-based learning in medical education: the Singapore hybrid. AB - INTRODUCTION: Problem-based learning (PBL) is an innovative educational strategy to enhance student-centred, interactive and integrated learning through intensive small group tutorials. PBL is problem-first learning in which a commonly encountered medical problem serves as the primary focus of and stimulus for learning. Several adaptations have evolved since its implementation at McMaster University in 1969. In 1999, the Singapore medical school implemented PBL as a key feature within its new hybrid curriculum. METHODS: The review will focus on the educational implications and impact of the present curriculum reform with emphasis on the implementation of PBL within the new hybrid medical curriculum. RESULTS: Our early experience in implementing PBL resembles those of several other "traditional" medical schools, namely, a generally favourable and positive response from staff and students mixed with varying degree of anxiety to change. CONCLUSION: Appropriate training and changing the mindsets of staff and students, strong leadership from the Dean and a deep commitment by all concerned are essential to ensure the successful implementation of PBL within a "traditional" medical school. PBL will enhance the quality of medical education and the educational preparation of our students to meet the challenges of future medical practice. PMID- 11503540 TI - Quality assurance of problem-based learning (PBL): the Hong Kong experience. AB - INTRODUCTION: The Faculty of Medicine of the University of Hong Kong launched its new medical curriculum in 1997. It was an integrated and system-based curriculum, with problem-based learning (PBL) being one of its new pedagogy. This paper describes the quality assurance of the introduction of PBL. MATERIALS AND METHODS: Regular PBL tutor training and case-writing workshops were conducted, with experts both locally and internationally invited to serve as trainers. The monitoring mechanisms was the use of paper questionnaires filled by both students and teachers. A group with a view to improve the process and the content of PBL studied the ratings of these evaluations. RESULTS: In 3 years, a total of 40 training workshops were held and, 268 tutors and 57 case-writers were initiated. Most of the teachers were convinced that PBL did work even in students who came from a didactic learning background. Extensive and continued refresher PBL workshops would be necessary. CONCLUSION: A stringent monitoring and evaluation process formed an integral part of an effective introduction of PBL in the medical curriculum. PMID- 11503541 TI - Assessment in problem-based learning: the role of the tutor. AB - INTRODUCTION: Problem-based learning (PBL) in medicine emphasises the tutor's role in facilitating collaborative and integrated learning. While it is widely recognised that tutors come to know their students, they traditionally play little part in the formal assessment process. We introduced tutor assessment into a problem-based medical curriculum and examined the patterns of tutor marks in four subjects, in terms of within semester consistency and their relation to other forms of assessment in the course. MATERIALS AND METHOD: Seventy-four tutors assessed 187 students twice (formative assessment in mid-semester, summative assessment at the end of semester) in each semester, using an assessment tool that focussed on communication and learning skills. Tutor marks were examined using correlations, cluster analysis and chi-square coefficients. RESULTS: There were consistent trends in the patterns of cluster membership for pairs of marks in high, average and low clusters from mid-semester formative to end-of-semester summative assessment. Cluster membership for pairs of marks was predominantly consistent. Most movement was from lower to higher clusters over the two assessments, reflecting student improvement after tutors' feedback and tutor responsiveness to feedback to them. There was little variance in the marks of different tutors. Tutor marks correlated moderately and significantly with other forms of assessment in each subject. CONCLUSION: Tutor marks contribute useful, distinctive dimensions to assessment in a problem-based medical curriculum. PMID- 11503542 TI - Introduction of problem-based learning in a traditional medical curriculum in Singapore--students' and tutors' perspectives. AB - OBJECTIVES: The Faculty of Medicine at the National University of Singapore was founded in 1905 and has trained many generations of medical practitioners. Teaching has been based on a traditional British-style curriculum with 2 years of training in the basic clinical sciences and 3 years in the clinical disciplines. Starting in the academic year 1999-2000, a more integrated curriculum was introduced. In conjunction with this, approximately one-fifth of the curriculum time was dedicated to problem-based learning (PBL). This will be the first time that PBL is being implemented in the medical school and both staff and students will be new at it. Thus, the objective of this study was to gather information on the reactions of both staff and students after the actual implementation. MATERIALS AND METHODS: A questionnaire was designed to assess the following: (1) What are tutors' and students' opinions on the relative benefits on students' learning process and participation of PBL versus traditional lectures? (2) What is the level of satisfaction with various aspects of their PBL experience? (3) What were the difficulties that were encountered? RESULTS: Several positive and negative aspects of the tutors' and students' experiences were revealed. Most reported fairly high levels of satisfaction with their PBL experience. CONCLUSIONS: Overall, the experiences have been positive and both groups are willing to "struggle" with this new way of learning. PMID- 11503543 TI - Encouraging learning how to fish: an uphill but worthwhile battle. AB - INTRODUCTION: Encouraging teaching practices such as problem-based learning (PBL) amongst undergraduate students within a lecture-based, system-based integrated curriculum is a challenge. Students are apprehensive about developing an organised framework for acquiring knowledge while lecturers are required to reframe their views on the educational process and their role as educators. MATERIALS AND METHODS: Lecturers and students in the Phase (Year) II programme were asked to fill questionnaires following the second and fourth PBL cases. The two sets of survey responses were compared to see whether the students' and teachers' perceptions had changed over the 5-month period. RESULTS: Students' responses from both surveys (1 and 2) were similar in that a majority agreed that the PBL tutorials had encouraged the seeking of information (66% and 67%, respectively), had improved understanding (57% and 56%), integration (65% and 70%) and application (50% and 64%) of knowledge. However, the views given in the form of written comments, following their positive responses, were somewhat contradictory. A large number of students (38% and 40%) faced difficulties in getting involved in discussions during the PBL tutorial and a majority (73% and 82%) preferred the normal subject-based tutorials. The reasons given by approximately 20% of the students were that the subject-based tutorials were more efficient for obtaining information and/or that the information had been pre selected by the lecturers. More than 80% of the lecturers (in both surveys) perceived that the students had identified the appropriate learning objectives and covered the subject matter. The percentage of lecturers who agreed that PBL tutorials encouraged rapport and teamwork amongst students had increased in the second survey, from 70% to 92% and 55% to 83% respectively. CONCLUSION: Implementing PBL is not simply a matter of developing new teaching materials and new effective ways of presenting them. It requires a paradigm shift, a change in the roles of students and teachers, and time. PMID- 11503544 TI - Students' perception of problem-based learning in the medical curriculum of the Faculty of Medicine, University of Colombo. AB - INTRODUCTION: Problem-based learning (PBL) is used as an instructional method in the system-based modules of the new innovative curriculum of the Colombo Medical Faculty in Sri Lanka. This study was undertaken to assess whether objectives of having PBL to facilitate acquisition of desirable learning skills and generic skills had been achieved. The perceived advantages and disadvantages of PBL were also studied. MATERIALS AND METHODS: The study population comprised third year medical students who had completed 12 PBL sessions. A self-administered questionnaire, which measured learning outcomes and acquisition of generic skills on a 5-point Likert rating scale, was used. RESULTS: The response rate was 87.2% (n = 188). Eighty per cent of students were aware of the rationale for having PBL and of learning from recommended material. Seventy-five per cent knew the competencies that could be acquired from PBL and two-thirds knew about the importance of small group discussions. PBL had helped to improve communication skills in 57% and problem solving skills in 52%. The main disadvantage was that it was time-consuming. Only 46.6% were satisfied with the participation of colleagues in the group. Seventy-six per cent felt that PBL could be better conducted. CONCLUSION: Awareness of concepts of PBL, reasons for its inclusion in the curriculum and educational advantage was high. The main disadvantage perceived was that it was time-consuming. There is a need to improve the conduct of PBL and, provide guidance and awareness programme for students. PMID- 11503545 TI - Problem-based learning (PBL) as an approach in the teaching of biochemistry of the endocrine system at the Angeles University College of Medicine. AB - INTRODUCTION: Biochemistry is a basic science subject introduced in the first year of the medical curriculum. At the Angeles University College of Medicine, the approach used in teaching biochemistry has always been the conventional lecture-based strategy. This study described the factors involved in the development of modules and the use of problem-based learning (PBL) as an innovative strategy in teaching the biochemistry of the endocrine system. MATERIALS AND METHODS: Four PBL modules consisting of clinical problems, student's and facilitator's guides and a list of learning objectives were developed and used by 68 first year medical students under the supervision of a tutor during small group tutorial sessions. A 5-point Likert scale questionnaire was employed to ascertain the perceptions of the students on the influences of the components of PBL on the learning process. RESULTS: The respondents affirmed that the PBL approach motivated them to actively control the direction of their learning needs and encouraged them to acquire self-learning skills. Ninety per cent of the students found the PBL approach to have inspired them to take charge of their own learning of the biochemistry of the endocrine system. CONCLUSION: The study showed that the students found this alternative method acceptable particularly in motivating to clarify biochemical concepts that facilitated their understanding of selected endocrine problems. PMID- 11503546 TI - Prevention of human immunodeficiency virus (HIV) transmission from mother to child--a cohort study in Singapore. AB - INTRODUCTION: The landmark Paediatric AIDS Clinical Trials Group (PACTG) trial 076 showed in 1994 that antiretroviral therapy (ART) was effective in reducing maternal-child transmission of human immunodeficiency virus (HIV). This trial included antenatal oral zidovudine (ZDV), intrapartum intravenous ZDV, 6 weeks of oral ZDV to the babies and no breastfeeding. MATERIALS AND METHODS: This study is an on-going, prospective, open-label trial conducted from 1995 in which we enrolled HIV-infected pregnant women using the above strategy. Since 1997, the antenatal component of the regimen was modified to include lamivudine with ZDV. All babies had serial HIV polymerase chain reaction (PCR) and antibody tests including enzyme-linked immunosorbent assay (EIA), particle agglutination (PA) and Western blot (WB) at day 1, 1 week, 1, 2, 3, 6, 12 and 18 months. RESULTS: A total of 16 out of 19 eligible women were recruited from 1995 to 1999. The median age was 26 years (range 22 to 38 years), 38% were Singaporeans, median CD4 was 421 cells/mL (range 18 to 713 cells/mL) and median baseline gestational age was 23.5 weeks (range 8 to 32 weeks). None of the 16 children was infected as evidenced by 2 negative HIV PCRs including 1 done > 4 months old with a follow-up of 6 months to 2 years. There was a statistically significant difference between the 3 HIV antibody tests at 12 months of age (P = 0.003), there being more negative results with WB as compared to PA (P = 0.02). However, the difference between the 3 tests at 18 months was not statistically significant. No long-term side effects in these children were seen. CONCLUSION: Although the number of patients in this study is small, the absolute prevention of transmission (95% confidence intervals 0%-17%) in this cohort supports the recommendation of antenatal HIV screening and treatment of those infected. PMID- 11503548 TI - Body mass index and its related factors in the elderly. AB - BACKGROUND: A cross-sectional study was conducted in May 1998 in Chung-Hsing Village, Taiwan to evaluate the relationship between body mass index (BMI) and its related factors in the elderly. METHODS: Individuals aged 65 years and over were recruited as study subjects. A total of 1093 persons, out of 1774 registered residents, were contacted for face-to-face interview (61.6%). However, only 586 respondents completed the questionnaire and had blood tests. Analysis was based on these 586 subjects. RESULTS: There were 66.0% men and 34.0% women. The mean age was 73.1 +/- 5.3 years. The BMI was 24.13 +/- 4.64 kg/m2 and 24.07 +/- 3.99 kg/m2 for men and women, respectively (P > 0.05). In univariate analysis, high systolic pressure, high diastolic pressure, hypertriglyceridaemia, hyperglycaemia and hyperuricaemia were related to obesity. After controlling for the other covariates, the multivariate logistic regression analysis showed that significant related factors of obesity were hyperglycaemia and hyperuricaemia. CONCLUSIONS: Significant related factors of obesity in the elderly were hyperglycaemia and hyperuricaemia. A large-scale investigation will be suggested in the future to address causal-effect issues between obesity and hyperglycaemia or hyperuricaemia. PMID- 11503547 TI - Body mass index profile in Hong Kong Chinese adults. AB - INTRODUCTION: Obesity has now become an epidemic with increasing prevalence in most parts of the world. However, information on the problem of obesity in Asians is limited. SUBJECTS AND METHODS: From April 1996 to August 1997, 17,121 Chinese adult subjects from the community of Hong Kong presented themselves voluntarily at the United Christian Nethersole Community Health Service Centers for primary health care check-up. Demographic data including height and weight were documented. Their body mass indexes (BMI) were measured. The prevalence rates of overweight and obese subjects were calculated according to the 1995 World Health Organization criteria. RESULTS: There were 4781 (27.9%) men and 12,340 (72.1%) women. The mean age (+/- SD) was 51.3 +/- 16.0 years (range 20 to 96 years, median 49.8 years). Of the 17,121 subjects, 4939 (28.85%) were overweight and 656 (3.83%) were obese. Based on the 1997 Hong Kong Census statistics of the overall Hong Kong population data, the age-standardised prevalence rates of overweight and obese subjects were 30.52% and 3.19% in men and 22.14% and 3.35% in women, respectively. Of the 10,937 subjects of working age (20 to 65 years), the prevalence rates of overweight and obese subjects were 28.98% and 3.55%, respectively. Of the 6648 subjects aged > or = 60 years, the prevalence rates of overweight and obese subjects were 36.31% and 4.81%, respectively. CONCLUSIONS: In Hong Kong, the age-standardised prevalence rate of overweight and obese subjects were 30.5% and 3.2% in men and 22.1% and 3.4% in women, respectively. This information provides some useful estimations of the extent of an important public health problem, obesity, in Hong Kong. PMID- 11503549 TI - Treatment of acute promyelocytic leukaemia using a combination of all-trans retinoic acid and chemotherapy. AB - INTRODUCTION: The combination of all-trans retinoic acid (ATRA) with chemotherapy has improved the outcome of acute promyelocytic leukaemia (APL). Effective induction as well as maintenance therapy for APL can be achieved using this combination of anti-leukaemic agents. MATERIALS AND METHODS: Twenty-four consecutive patients with newly-diagnosed APL were treated with ATRA daily together with either daunorubicin or idarubicin. Therapy with ATRA was continued until complete remission (CR) was achieved; thereafter, patients were treated with 2 cycles of an anthracycline-based consolidation chemotherapy (either daunorubicin or idarubicin). Maintenance therapy was achieved using 5 alternating cycles of low-dose methotrexate (MTX) plus 6-mercaptopurine (6MP) followed by ATRA alone. RESULTS: Twenty-three out of 24 patients (96%) completed induction therapy and achieved haematological CR (HCR) as well as molecular remission (MR); however, 1 patient (5%) died from retinoic acid syndrome. Twenty-one out of 23 evaluable patients (91%) completed consolidation chemotherapy, and 2 patients (10%) died, 1 from neutropenic sepsis and the other from relapse following non compliance to therapy. All 21 surviving patients in the present study received maintenance chemotherapy and are still in HCR and MR at a median follow-up of 23 months. The estimated actuarial 2-year overall survival (OS) and event-free survival (EFS) rates were both 84% +/- 9%. CONCLUSION: The combination of ATRA with an anthracycline is an effective remission-induction therapy for newly diagnosed APL. Maintenance therapy using alternating cycles of MTX plus 6MP followed by ATRA alone is effective in maintaining CR and MR as well as prolonging the survival of patients with APL. PMID- 11503550 TI - A retrospective study of incontinentia pigmenti seen at the National Skin Centre, Singapore over a 10-year period. AB - INTRODUCTION: Incontinentia pigmenti is a rare X-linked dominant disease which affects the ectodermal tissues, usually lethal in males. MATERIALS AND METHODS: A retrospective analysis of clinical data obtained from the photographic documentation and casenotes of patients diagnosed to have incontinentia pigmenti at the National Skin Centre. The study covered the period from January 1990 to December 1999. RESULTS: Twenty-six patients were diagnosed to have incontinentia pigmenti of the Bloch-Sulzberger type; 23 (88.5%) were females and 3 (11.5%) were males. There were 20 Chinese, 3 Malay and 3 Indian patients. Most patients had cutaneous manifestations at birth or within the first week of life. Cutaneous features included vesicles, papules, verrucous plaques and splash-like hyperpigmentation along the lines of Blaschko. The cutaneous lesions were widespread in 21 (81%) and localised in 5 (19%) patients. In some cases, hypopigmented atrophic streaks (2 patients) or whorled scarring alopecia (4 patients) were seen. Extracutaneous manifestations, seen in 5 (19%) patients, included neurological, dental and ocular defects. One Malay girl had severe neurological involvement associated with ocular abnormalities. A positive family history was present in 6 (23%) patients. The 3 male patients were Chinese without any family history. CONCLUSIONS: Each stage of the disease comes with its own set of differential diagnosis, including infections e.g. herpes virus infection and other types of genodermatoses e.g. linear and whorled nevoid hypermelanosis. The phenomenon of whorled scarring alopecia, hitherto unreported in the literature, corresponded to the lines of Blaschko. In the 3 Chinese male patients, the disorder probably originated from a new mutation. X chromosome inactivation in females during early embryogenesis results in a mosaic population of cells and this explains the linear and patchy manifestations of incontinentia pigmenti. PMID- 11503551 TI - Malignant melanoma seen in a tertiary dermatological centre, Singapore. AB - INTRODUCTION: Primary malignant melanoma is an uncommon skin tumour in Singapore compared with the Western countries. The clinical characteristics of melanoma have been rarely reported in Asians. OBJECTIVES: Our purpose was to study the incidence, clinical characteristics and risk factors of biopsy proven malignant melanoma seen in a tertiary referral skin hospital. METHODS: Case records of patients with histology proven malignant melanoma from January 1989 to December 1998 were retrieved. Patients were interviewed and a complete skin and systemic examination was performed. Data on demographics, histological types, clinical characteristics and risk factors were collated. RESULTS: Between 1989 and 1998, 27 patients were diagnosed with histology proven malignant melanoma. There was a predominance of Chinese with a female to male ratio of 1.3:1. There was a mean lag period of 1.6 years to diagnosing melanoma and there is a general lack of knowledge among the local patients. Most of the lesions (89%) occurred on the extremities. Acral lentiginous melanoma (41%) and nodular melanoma (41%) were the two commonest histologic types. Of the nodular melanomas, 64% were observed in patients below 50 years (64%) while majority of the acral lentiginous melanomas occurred in those above 50 years (91%). In terms of risk factors, we reported a high prevalence of diabetes mellitus with acral lentiginous melanoma. CONCLUSIONS: This study highlighted interesting aspects in terms of clinical characteristics and risk factors in our local patients. It also served as a remainder of the need to raise awareness among the public and medical community of skin cancers or melanomas as they are potentially treatable if detected early. PMID- 11503553 TI - Case reports of low dose cyclosporine. A therapy in adult minimal change nephrotic syndrome. AB - INTRODUCTION: Many centres still use steroids to induce remission in patients with minimal change nephrotic syndrome (MCNS) and failing that to give a course of cyclophosphamide, though some centres are already using cyclosporine A (CsA) as an alternative. We report the benefits of CsA therapy in 3 adults with difficult to treat MCNS in whom low dose CsA therapy proved to be efficacious. CLINICAL PICTURE AND OUTCOME: The first patient had her 1st relapse after 8 years and thereafter had 2 more relapses, within 3 months of each other, in spite of therapy with cyclophosphamide. With CsA therapy, at a dose of 3.5 mg/kg body weight (BW)/day, she achieved lasting remission of 22 months as of September 1999 and is still in remission. The second patient had his relapses of nephrotic syndrome over a period of 10 years when treated with prednisolone and cyclophosphamide. On the 13th relapse, he achieved a remission lasting 21 months after a 3 month course of CsA at a dose of 4 mg/kg BW/day. With the 14th relapse, he took half the dose of CsA prescribed [only the morning dose of neoral CsA (2 mg/kg BW/day)] and still achieved a remission and has been in remission since. The third patient was a young woman, married for 2 years without children. She could not tolerate prednisolone because of erosive gastritis and she responded to a pulse dose of intravenous cyclophosphamide for her 1st episode of nephrotic syndrome with complete remission. However, when she relapsed 5 months later she did not respond to a similar dose of i.v. cyclophosphamide and was therefore treated with CsA (4 mg/kg BW/day) which induced a prompt remission 1 month after commencement of therapy and she is still in remission. The trough CsA levels for the 3 patients (range 41 to 107 ng/mL) and the calculated average CsA levels were lower than that used for post renal transplant immunosuppression. The trough CsA levels were, however, similar to that used in patients with MCNS from other series, though achieved at lower CsA doses. CONCLUSION: Our study shows that low dose CsA is a useful agent for induction of remission of MCNS and maintenance of lasting remission. A low dose CsA regimen will make CsA more affordable. PMID- 11503552 TI - Catheter-related infection: diagnosis, prevention and treatment. AB - INTRODUCTION: Catheter infection continues to be a serious problem in critically ill patients. This review will examine the incidence, epidemiology, microbiology, diagnosis, risk factors for and treatment of catheter-related bloodstream infections. METHODS: Relevant articles were culled from a Medline search and other review articles on catheter-related infection. Important abstracts presented within the past year were included in the review if the data had not been published in a peer-reviewed journal. RESULTS: Catheter-related bloodstream infections (CRBSI) increase morbidity and the cost of care of patients. The predominant organisms associated with CRBSI are coagulase negative staphylococci, enterococci and Staphylococcus aureus. Diagnosis usually requires catheter removal and culture but newer techniques such as "differential time to positivity" may permit diagnosis of CRBSI in situ. Reducing risk factors for infection are important: education of house staff, use of the subclavian insertion site, skin preparation with chlorhexidine solutions, use of maximum barrier precautions during catheter insertion and catheter maintenance with chlorhexidine sponges. If infection rates are still high after institution of these measures, use of antiseptic or antibiotic-impregnated catheters should be considered. Treatment of CRBSI in critically ill patients mandates catheter removal and treatment with systemic antibiotics. CONCLUSIONS: The best treatment for CRBSI is prevention. Ongoing studies are evaluating the effectiveness of newer antiseptic catheters, the risk of developing antibiotic resistance when using antibiotic-impregnated catheters and the contribution of multiple interventions on the development of CRBSI. PMID- 11503555 TI - Clinical update on Helicobacter pylori. PMID- 11503554 TI - A rare cause of syncope in a patient with diabetes mellitus--a case report. AB - INTRODUCTION: Hypoglycaemic episodes in patients with diabetes mellitus are mostly due to excess doses of exogenous insulin or oral hypoglycaemic agents, coupled with poor caloric intake and excessive unplanned physical exertion. Hypoglycaemia as a result of endogenous hyperinsulinaemia due to an insulinoma is extremely rare in such patients. CLINICAL PICTURE: This patient with type 2 diabetes mellitus presented with episodes of syncope. Investigations confirmed recurrent hypoglycaemia from endogenous hyperinsulinaemia, with localisation of a tumour in the tail of the pancreas. TREATMENT: Distal pancreatectomy and splenectomy. Histology confirmed an insulinoma. OUTCOME: No further hypoglycaemic episodes were noted. The patient returned to his diabetic state with rather poor glycaemic control. CONCLUSIONS: Repeated hypoglycaemic episodes in a patient with diabetes mellitus despite complete withdrawal of hypoglycaemic agents should lead one to consider other causes of hypoglycaemia. PMID- 11503556 TI - Introduction to Bayesian methods for medical research. AB - Bayesian statistical methods provide a credible alternative to traditional statistical approaches used in medical research. In this article, the author gives a brief introduction to the key concepts involved in Bayesian methods and compares them with some commonly used non-Bayesian statistical techniques. PMID- 11503557 TI - The 4th Tan Tock Seng Hospital Oration: challenge for the medical profession in the 21st century. PMID- 11503558 TI - [Clinical practice improvement--clinical practice quality improvement of local health care processes]. AB - Clinical Practice Improvement (CPI) is a methodological approach to develop analytically-based protocols to achieve desirable outcomes at the lowest essential cost over the continuum of local care processes. Several elements of the CPI approach make it attractive to clinicians: First, it is a scientific bottom-up approach that places accountability for practice improvement and outcomes with local clinicians. Clinicians are not told to blindly follow a guideline or protocol developed by others, but instead collect data on outcomes, on treatments, and on patient signs and symptoms that support practice change. CPI supports caregivers in making their own decisions about optimal care on the basis of objective statistical evidence gathered in the routine, everyday practice of medicine. Second, CPI measurement encompasses a comprehensive view of the care management process: patient characteristics, process steps, and outcomes. All three classes of data are considered simultaneously. This comprehensive measurement framework provides a basis for meaningful analyses of significant associations, as well as relationships between process and outcome. Third, the CPI methodology focuses on deployment and application. There is a continual emphasis on factors that can be implemented to improve outcomes and the process to achieve these results. This focus on implementation guides who is involved in the design, what data are collected, what questions are answered during analyses, and who designs the protocols or improvements in practice. PMID- 11503559 TI - [Evaluation of a patient-held medical record by patients--results from the Ried Medical Quality Network]. AB - In the "medical-quality network Ried", an independent practice network in the south of Hessia, an A 4 format patient held record for multi-morbid chronically ill patients is used since 1997. 655 patients (response-rate 71.0%) from sixteen general practitioner (GP) practices were surveyed by written questionnaire. Respondents were in average 68.1 years of age, and for 16.3 years with their GP. 52.2% were male. Patients had their own records for 24.2 months and had in average 1.9 (women) or 2.3 (men) serious health problems. Women had in average 5.0, men had 4.8 different drugs prescribed. 96.7% of the respondents thought that the patient-held record is a good idea, 85.5% felt more secure and 98.7% have it always at hand at home. The patient-held record provides better information about ones own health (92.1%) and 96.6% want to be able to look at their own record. The patient-held record makes it easier for patients to talk to doctors (86.6%) and provides a better overview for the GP (90.8%). The more drugs patients have to take, the more important it becomes for them to have their own record. 68.6% take the record on holyday, 57% take it to other doctors than the GP. 76.4% reported that the patient-held record was used at least once by a specialist, 61.3% by a hospital. 21.8% remembered emergency situations where it was helpful. Only 4.8% thought that it is tiresome. Few proposals for improvement were made, mainly focussing on a smaller format. On the basis of these results patient-held records should be a core activity of practice networks, especially for this type of patients. PMID- 11503560 TI - [Evidence-based medicine in internal guideline development in a general hospital- the Park-Clinic EbM-Project]. AB - Evidence-based medicine (EbM) is despite its historical roots still a young concept that is quite relevant in everyday work, but yet little used. The first hospital specific approach to implement EbM into the health care system and daily clinical practice in Germany is recorded in the project of the Park-Klinik Weissensee (Berlin). Implementation and execution of EbM into daily routine should be reached through five steps which should result in better quality of treatment of the patients: 1. EbM-Training for hospital staff; 2. EbM-Search (= access to electronic databases and EbM-information); 3. Guideline-Committee (= hospital committee for the development of clinical practice guidelines); 4. EbM Recommendation (= use of EbM-information in discharge letters) and, finally 5. EbM-Events (= vocational trainings for hospital physicians and general practitioners). Although project duration is yet quiet short, first results allow to look into the future with confidence. At the same time it can be recognized how long and difficult the way for all participants will be to reach evidence based information and communication. The "Park-Klinik EbM-Project" can make an important contribution to health care services research in Germany. PMID- 11503561 TI - [Interviews with physicians in private practice as a starting point for quality improvement in hospitals]. AB - Subjective criteria gain importance in care research and quality management. Therefore, psychiatrists in private practice and general practitioners working in the catchment area of a psychiatric hospital were surveyed by questionnaire evaluating expectations and satisfaction concerning their collaboration with the hospital. Psychiatrists and general practitioners rated legible interim discharge letters, good in-patient treatment, adequate diagnostics, and direct referral to the hospital as most important. Psychiatrists express dissatisfaction regarding prescription of expensive drugs, lack of involvement in planning new psychiatric institutions, delivery of discharge summaries, referring back the patient after in-patient care, and delay of hospital admission. As conclusion, quality management should pay more attention to ambulatory care physicians' points of view to reduce problems of interaction between clinicians and their colleagues in private practice and to improve the treatment continuity of psychiatric patients. PMID- 11503562 TI - [Innovations in medicine--the structured way from an idea to recognized treatment]. AB - New techniques in medicine in an enormous variety and limited financial resources make it necessary to define step by step the "national medical how to do". This is to be done by the four boards of health care self-government--especially by the "Koordinierungsausschuss". Thus the clinically active physicians are enabled to implement their own new ideas and their own innovations--especially via high quality guidelines, consented within the scientific societies. This new closed circle, a kind of "government free zone" should be used intensively by all active colleagues. PMID- 11503563 TI - [Modern quality management in pathology--accreditation according to EN DIN 45004: an experience report]. AB - Accreditation according to EN DIN 45004 is a new way of documenting quality assurance and competence in pathology. It comprises a quality assurance program according to ISO 9002 as well as demonstration of diagnostic competence to a panel of experts. Accreditation results in a marked improvement of diagnostic quality and appears suitable to document special competence in pathology according to a flexible internationally recognized norm. We recommend accreditation according to EN DIN 45004 to all competent institutes of pathology. PMID- 11503564 TI - [Importance and possibilities of general guidelines in family practice strategies -do we need 'basic guidelines' in general practice?]. AB - General practice is distinguished particularly by complex patient problems and generic competencies to act and counsel as a family practitioner. Guidelines which are only centered on clinical topics like diagnoses, treatments etc. are not sufficient to support action and decision in general practice. The article proposes the concept of generic guidelines (basic guidelines) for family medicine. A need for basic guidelines in general practice is established in three areas: 1) cross sectional patient problems (e.g. care for immigrants, common strain and distress, counselling of screening procedures), 2) basic skills and strategies in family practice (e.g. problem-oriented consultation, home visits, patient information and informed consent), and 3) practice management and documentation. The development of generic guidelines seems to be difficult as the efforts to evidence-based professional practice are only recently started, but this is matched by the high importance of guidance for general practice, and certain concepts of family medicine support it. It is concluded that the development of basic guidelines in general practice will have a considerable impact on structuring the framework and fostering the quality improvement of general practice. PMID- 11503565 TI - [Problems in quality assurance in physiotherapy of rheumatic diseases]. PMID- 11503566 TI - Wireless LANs in healthcare. AB - Wireless local area networks (WLANs) function much like conventional LANs, except that the information they carry is transmitted using radio-frequency (RF) signals and sometimes infrared signals rather than copper or fiberoptic cables. Using devices that communicate over a WLAN has several benefits: For example, the devices can be used virtually anywhere in the facility without needing access to a hardwired data terminal, and they are often easier to install since they don't need additional wiring. But WLANs also have drawbacks. They can't transfer as much data at one time as wired systems, for example, or transfer it as quickly. Because they require new technology, they may carry greater costs. And because they operate in a frequency band that isn't protected from interference, they require special techniques to maximize the quality of their signals--techniques that, themselves, require a number of challenging decisions. In this article, we describe the basic operating and purchasing considerations involved in deciding whether to invest in WLAN technology and which type of system to choose. PMID- 11503567 TI - Continuous renal replacement therapy. The real story. AB - Continuous renal replacement therapy (CRRT) is used to treat patients who suffer from acute renal failure. Unlike the more traditional intermittent hemodialysis (IHD), CRRT is administered around the clock, providing patients with nonstop therapy and sparing them the destabilizing hemodynamic and electrolytic changes characteristic of IHD. Many clinicians have reported that CRRT's steady, milder treatment is less traumatic for some of their patients than IHD. For acute renal failure patients who can't tolerate IHD--usually those with very low blood pressure--CRRT is often the only choice of treatment. CRRT can also be used on many patients who can tolerate IHD. But because there is little useful survival data concerning CRRT, and because CRRT is sometimes perceived as significantly more expensive than IHD, some hospital administrators have--groundlessly, we believe--questioned the wisdom of investing in this technology. In this article, we examine CRRT's benefits and take a close look at some of the myths that have impeded its adoption. PMID- 11503568 TI - Tara KLamp circumcision clamp. PMID- 11503569 TI - Electromagnetic interference from linear accelerators can affect electronic devices. PMID- 11503570 TI - Ventilator power cords are lifelines that need careful inspection. PMID- 11503571 TI - Shorter-than-expected battery operation time for MDE prism modular physiologic monitors. PMID- 11503572 TI - Potentially defective power supply boards in certain Nellcor pulse oximeters. PMID- 11503573 TI - [Generalized convulsive status epilepticus]. PMID- 11503574 TI - [Generalized convulsive status epilepticus: analysis apropos of 57 cases]. AB - BACKGROUND: The generalized convulsive status epilepticus (GCSE) is a medical emergency with a high morbimortality rate. We analyzed the casuistry in our hospital and we assessed the causes related to mortality. PATIENTS AND METHODS: This is a retrospective study of 57 cases of GCSE that took place between january 1992 to december 1998. The variables studied were mortality among groups according to age, previous epilepsy and the duration of the convulsive episode. RESULTS: We found male prevalence (63.2%). The 49.2% of patients didn't have previous history of epilepsy. In 38.6% of cases the treatment dose was insufficient and in 61.4% it was delated. The mortality rate was 36.8% with a higher significance in patients older than 65 (p < 0.02), without previous history of epilepsy (p < 0.001) and in episodes lasting more than 4 hours (p < 0.05). Only 12.9% of patients received prehospital treatment. CONCLUSIONS: The mortality of GCSE is very high. The initial treatment is inadequate. We must establish a premature and adequate protocol, with optim pharmacological doses and a good coordination with intensive care units. PMID- 11503575 TI - [Pattern of blood levels of erythropoietin and proinflammatory cytokines in patients with anemia of chronic disorders secondary to infection]. AB - OBJECTIVE: To evaluate the pattern of erythropoietin (EPO) and some proinflammatory cytokines in the anemia of chronic disorders (ACD) secondary to infection. METHODS: Sequential determination in serum of interleukin-1 beta (IL-1 beta), necrosis tumoral factor alpha (TNF-alpha), gamma interferon (IFN-gamma), interleukin-6 (IL-6), and erythropoietin (EPO) in 25 patients with chronic bacterial infectious diseases and ACD criteria. We evaluated the relationship of these mediators with the anemia and the iron metabolism. RESULTS: Serum EPO levels significatively decreased compared with initial values, and the last control was in normal rank (18.04 +/- 19.10 vs. 8.56 +/- 4.72 UI/mL; p < 0.001; normal rank: 4-15 mUI/mL). In the first control, there was a negative and non significative correlation between the EPO levels and the hemoglobin concentration (r = -0.115, NS), reaching significance in the last control (r = -0.446; p < 0.05). There was negative correlation between the hematocrit and TNF-alpha levels (r = 0.467; p < 0.05) and between the haemoglobin values and the log of serum TNF alpha (r = 0.424; p < 0.001). An inverse correlation between the IL-6 levels and both, the hemoglobin concentration and the serum iron was found, and there was a direct correlation between this cytokine values and the EPO levels. CONCLUSIONS: Blunted response of erythropoietin and the action of TNF may contribute to the pathogenesis of ACD secondary to infection. Positive correlation between IL-6 and EPO suggest a proerythropoietic action of IL-6 in response to the anemia. PMID- 11503576 TI - [Ambulatory blood pressure monitoring in young patients]. AB - OBJECTIVE: To defining the criteria for performing ambulatory blood pressure monitoring (ABPM) in young patients. METHOD: It is reported the experience with ABPM on 52 consecutive patients (younger than 30 years old) consulting for hypertension (mean age 23.4 +/- 4.9 years). The ambulatory BP was measured noninvasively for twenty-seven hours by the Spacelabs 90207 device programmed to measure BP every fifteen minutes during daytime and every 20 minutes during nighttime. The definition of daytime and nighttime was made on the basis of wakefulness and sleep or bed rest periods, obtained from a diary kept by the subject, normal nocturnal BP drop was defined as a decrease higher of 10% versus daytime values. It was defined normal BP an 24 hours ambulatory BP < 130/80 mmHg. RESULTS: Thirty seven patients (71%) were normotensives. There were not differences between normotensive and hypertensive patients neither by age (normotensive 23.9 +/- 12.5, hypertensive 23.3 +/- 4.0 years), nor by sex (normotensive, 21 men and 16 women; hypertensive, 10 men and 3 women). Mean 24 h BP of normotensive patients was 119/72 mmHg (p < 0.001 vs. hypertensive, 135/89 mmHg). There were not differences in nocturnal BP drop. White-coat reaction was more intense in normotensive patients (1.17 +/- 0.12, vs. hypertensive 1.04 +/- 0.08, p < 0.001). Four hypertensives showed white-coat reaction (1.11 +/- 0.05). CONCLUSION: ABPM is a helpful diagnostic tool in young patients. It should be routinely performed as first exploration in all patients younger than 30 years consulting for hypertension. PMID- 11503577 TI - [Differences between tuberculous spondylitis and brucellar spondylitis]. AB - OBJECTIVE: To identify potential differences in the clinical and laboratory characteristics between tuberculous spondylitis (TS) and brucellar spondylitis (BS). PATIENTS AND METHODS: Retrospective study of patients with TS and BS diagnosed in our hospital between january 1992 and december 1998. RESULTS: TS was diagnosed in 17 patients and BS in 10 patients. In our series, a higher delay in the diagnosis (27.9 +/- 24.6 vs. 16 +/- 5.6 weeks, p = 0.02) was found in TS. There was a higher frequency, but without stadistic significance, of immunosuppression, one or several paravertebral or epidural abscesses, spinal cord compression, anemia and an elevated erythrocyte sedimentation rate in TS, and a higher frequency of fever/febricule and residual vertebral pain in BS. Lumbar location was the most frequent in both groups (58.8% in TS and 70% in BS). CONCLUSIONS: It is possible that there were some differences in the clinical and laboratory characteristics between TS and BS which may be an aid in the differential diagnosis of both entities and orient the empirical treatment in these cases without a definitive microbiological diagnosis or while awaiting the diagnostic confirmation. PMID- 11503578 TI - [Varicella pneumonia in the adult. Study of 9 cases]. AB - BACKGROUND: In the adult, the primary infection by the varicella-zoster virus acquires an unusual severity due to several complications, the most frequent of them being pneumonia. We study the main characteristics of nine patients diagnosed of pneumonia varicellosa. METHODS: Clinical, therapeutic and evolutive features of 9 adult patients, both immunocompetents and immunodepressed, diagnosed of pneumonia varicellosa are retrospectively reviewed, in the last ten years, at Hospital de Sant Pau, Barcelona. Diagnosis of varicella was established on the basis of the typical rash in the context of a feverish illness. The antecedents of smoking habit, pregnancy and underlying disease, evaluating especially arterial blood and platelet count at entrance, are assessed. RESULTS: Nine patients (4 males and 5 women; mean age 38 years) were included in the study. Seventy-eight percent of patients were smokers of more than 20 cigarettes a day; one met criteria of simple chronic bronchitis, another suffered ankylosing spondylitis and three were known carriers of human immunodeficiency virus. None of the female patients was pregnant. Respiratory symptoms began from the third and fifth day after the skin rash, and the most common symptoms were cough (89%), dyspnea (67%) and hemoptysis (22%). Arterial blood gas determination showed hypoxemia in four patients (45%). Chest X-ray revealed an interstitial pattern predominantly at both bases, with a case of right pleural effusion. Intravenous acyclovir was started in 6 patients, foscarnet in one and symptomatic therapy in two patients. All patients had a favourable clinical course, none of them requiring entrance to the Intensive Care Unit. CONCLUSIONS: Adult patients with varicella pneumonia that suffer respiratory insufficiency, thrombocytopenia or are carriers of base illnesses must be early treated with intravenous acyclovir. However, despite clinical, biological and radiological recovery is earlier with such treatment, the evolution seems equally favourable if it is only conducted, for instance, symptomatic therapy with antithermic and antihistaminic compounds. PMID- 11503579 TI - [Streptococcus suis meningitis]. AB - Human infection by Streptococcus suis (S. suis) is a zoonosis, with a known occupational risk and clinical presentation mainly as a purulent meningitis with low mortality and frequent hearing loss and ataxia sequela. Less than 150 human cases have been reported since original one thirty years ago. There is a geographical distribution most patients living in northern Europe and south Asia. S. suis disease in human has been reported in two patients in Spain the last years. We present two patients with S. suis meningitis, both were men with occupation related by pork meet, and good outcome. They come at our hospital in a lapse of one month. Both had neurosensorial hearing loss and walking ataxia. One patient had peripheral facial paralysis and diplopia because of paresia of contralateral sixth nerve, with complete resolution at three months. The rare presentation of S. suis meningitis in our country must not forget us to record the working risk at anamnesis. PMID- 11503580 TI - [Intravascular lymphomatosis manifesting in the lung]. AB - A case of intravascular lymphomatosis with predominant symptoms in the lung (cough and interstitial pulmonary infiltrates) is presented. Fever and confusion were lateness symptoms. Examination postmortem established the diagnosis. In the lung alveolar walls, small arterioles and the capillaries were occupied by atypical cells positives for CD45 and CD20, leukocyte and pan-B markers, and negatives for CD45-Ro and CD34, pan-T and endothelial markers. The main characteristics and the lung participation of this entity were reviewed. PMID- 11503581 TI - [Right ventricular cardiomyopathy: a diagnosis to bear in mind]. AB - Arrhythmogenic right ventricular cardiomyopathy is a entity of unknown etiology, that is pathologically characterized by right ventricular myocardial atrophy and fibroadipous tissue replacement. We present the case of a 65-year-old male patient with the diagnosis of chronic liver disease, whose study lead a diagnosis of Arrhythmogenic right ventricular cardiomyopathy. PMID- 11503582 TI - [Oxidative stress, diseases and antioxidant treatment]. AB - Oxidation is a biochemical process of loss of electrons associated with another of reception called reduction. This process is capital for life, because it takes part in the production of cellular energy. Oxidative stress appears when oxidation is excessive. This reality is complex in all biological levels, and cannot be measured or defined by a single parameter. A great number of diseases have been related to oxidative stress and generation of free radicals. For this reason, antioxidant therapies and diets (such as mediterranean diet) rich or enriched with antioxidants seem to prevent or at least to attenuate the organic deterioration originated by an excessive oxidative stress. PMID- 11503583 TI - [Giant cell pulmonary carcinoma in a patient with HIV infection]. PMID- 11503584 TI - [Bone metastasis as initial manifestation of follicular carcinoma of the thyroid]. PMID- 11503585 TI - [Endobronchial hamartoma: a case report]. PMID- 11503586 TI - [Genital tuberculosis and HIV infection]. PMID- 11503587 TI - [Patient with hypercalcemia and pancreatic islet cell tumor (parathyrinoma)]. PMID- 11503588 TI - [Late diagnosis of lepromatous leprosy in a geriatric patient]. PMID- 11503589 TI - [Disseminated intravascular coagulation secondary to acute brucellosis]. PMID- 11503590 TI - [Thrombopenia associated with mixed connective tissue disease and laparoscopic splenectomy]. PMID- 11503591 TI - [Cutaneous vasculitis and drugs]. PMID- 11503592 TI - [Use of AEP (Appropriateness Evaluation Protocol) in the emergency department of a hospital]. PMID- 11503593 TI - [Myopathy in hypothyroidism]. PMID- 11503594 TI - [Splenic marginal lymphoma: diagnosis made by splenectomy]. PMID- 11503595 TI - New England's Blue Cross Blue Shield shake-out: a case study in consumer activism. AB - For two generations, Blue Cross Blue Shield (BCBS) insurance plans could be counted on to play a key role in the financing of every community's health care system. "The Blues" dominated the health insurance market, yet they were also a reliable "insurer of last resort." In recent years, BCBS plans have begun to re structure to stay competitive. Four of New England's BCBS plans have proposed or completed mergers with Anthem Insurance, a mutual insurance company based in Indiana. This issue of States of Health looks at how advocates in New England are working together to protect health care consumers amid this transformation. PMID- 11503596 TI - Linking grassroots leadership and legal advocacy: partnerships for health care justice. AB - Every community has suffered its bout of health care turmoil. HMOs that suddenly announce they're dropping Medicare plans for seniors. Prescription drug costs that soar unabated. Increasing numbers of people without insurance despite a booming economy. Hospital shutdowns. News that the Blue Cross plan, long a community institution, is going for-profit. Because such changes affect us all, consumers must be at the table when decisions about the health care system get made. The need to strengthen the consumer voice--to institutionalize that voice and make it more effective--is prompting many consumer advocates to partner with attorneys. This States of Health looks at such partnerships and their tremendous potential for protecting consumers--and for achieving the broader goal of health care justice. PMID- 11503597 TI - Expanding Medicaid to cover the uninsured. AB - A decade of unparalleled economic prosperity has done little to reduce the number of Americans without health insurance. Despite an expansion of government-funded health insurance for the nation's low-income children, the number of working parents without health insurance continues to rise. Many states are looking at how Medicaid, long the health insurance program for families on welfare, could provide a health insurance safety net for the working poor. This issue of States of Health summarizes the issue and explores the Medicaid expansion strategies some states are using to provide health coverage for the working uninsured. PMID- 11503598 TI - [Interferon alfa-2a in the treatment of hepatitis C]. PMID- 11503599 TI - [Multimedia hospital in operation]. PMID- 11503600 TI - Licensing exams. PMID- 11503601 TI - Evidence-based psychiatric practice: implications for education and continuing professional development. Canadian Psychiatric Association position paper. PMID- 11503602 TI - Ask sirenhead. Care for the slow-drowning patient. PMID- 11503603 TI - Posterior leukoencephalopathy syndrome. PMID- 11503604 TI - Smoking and diabetes in Chinese men. PMID- 11503605 TI - Attachment of endometrial cells to peritoneum--facilitated? PMID- 11503606 TI - Impaired natural killer cell activity in endometriosis?--A technical challenge for validation. PMID- 11503607 TI - Risks of multiple pregnancy--the decision belongs to whom? PMID- 11503608 TI - Risks of multiple pregnancy--the decision belongs to whom? PMID- 11503609 TI - Conditions to perform endometrial laser intrauterine thermotherapy. PMID- 11503610 TI - Validity of comparisons? PMID- 11503611 TI - Comment on: Granulocyte macrophage-colony stimulating factor (GM-CSF) and sucralfate in prevention of radiation-induced mucositis: a prospective randomized study. PMID- 11503612 TI - In regard to Glatstein: Scientific physicians and evidence-based medicine. IJROBP 2001;49:619-21. PMID- 11503613 TI - In regard to Archambeau et al., IJROBP 2000;48:1155-1166. PMID- 11503614 TI - A cost-outcome analysis of adjuvant postmastectomy locoregional radiotherapy in high-risk postmenopausal breast cancer patients. PMID- 11503615 TI - Intra-arterial cisplatin more effective than intravenous. PMID- 11503616 TI - [Mitral subvalvular ventricular aneurysm]. PMID- 11503617 TI - Humeral lesion in a 69-year-old woman. PMID- 11503618 TI - The FDA and The Lancet: an exchange. PMID- 11503619 TI - The FDA and The Lancet: an exchange. PMID- 11503620 TI - The FDA and The Lancet: an exchange. PMID- 11503621 TI - The FDA and The Lancet: an exchange. PMID- 11503622 TI - The FDA and The Lancet: an exchange. PMID- 11503623 TI - Geographical differences for pneumococcal disease. PMID- 11503624 TI - Velocardiofacial syndrome or DiGeorge's anomaly. PMID- 11503625 TI - Direct observation for tuberculosis treatment. PMID- 11503626 TI - Direct observation for tuberculosis treatment. PMID- 11503628 TI - Medical education. PMID- 11503627 TI - Direct observation for tuberculosis treatment. PMID- 11503629 TI - Medical education. PMID- 11503630 TI - Oral transmucosal fentanyl and sufentanil for incident pain. PMID- 11503631 TI - Pamidronate in incident pain due to bone metastases. PMID- 11503632 TI - Efficacy of haloperidol in the treatment of nausea and vomiting in the palliative patient: a systematic review. PMID- 11503633 TI - Postural hypotension blamed on epidural. PMID- 11503634 TI - Re: Reversible delirium during opioid switching. PMID- 11503635 TI - Parks and factors in their success. PMID- 11503636 TI - Parks and factors in their success. PMID- 11503637 TI - In defense of antisense. PMID- 11503638 TI - Lateral gene transfer or viral colonization? PMID- 11503639 TI - Supplemental Security Income: determining disability for a child under age 18. Social Security Administration. Final rules. AB - On February 11, 1997, we published interim final rules with a request for comments to implement the Supplemental Security Income (SSI) childhood disability provisions of sections 211 and 212 of Public Law (Pub. L.) 104-193, the Personal Responsibility and Work Opportunity Reconciliation Act of 1996. We are now publishing revised final rules in response to public comments. We are also conforming our rules to amendments to Public Law 104-193 made by the Balanced Budget Act of 1997, Public Law 105-33. Finally, we are simplifying and clarifying some rules in keeping with the President's goal of using plain language in regulations. PMID- 11503640 TI - Food labeling: health claims; plant sterol/stanol esters and coronary heart disease. Food and Drug Administration, HHS. Interim final rule. AB - The Food and Drug Administration (FDA) is authorizing the use, on food labels and in food labeling, of health claims on the association between plant sterol/stanol esters and reduced risk of coronary heart disease (CHD). FDA is taking this action in response to a petition filed by Lipton (plant sterol esters petitioner) and a petition filed by McNeil Consumer Healthcare (plant stanol esters petitioner). Based on the totality of publicly available evidence, the agency has concluded that plant sterol/stanol esters may reduce the risk of CHD. PMID- 11503641 TI - Nephrology. PMID- 11503642 TI - Extension of expiration date for the respiratory body system listings. Social Security Administration (SSA). Final rule. AB - We adjudicate claims at the third step of our sequential evaluation process for evaluating disability using the Listing of Impairments (the Listings) under the Social Security and Supplemental Security Income (SSI) programs. This final rule extends until July 2, 2002, the date on which the respiratory body system listings will no longer be effective. We have made no revisions to the medical criteria in these listings; they remain the same as they now appear in the Code of Federal Regulations. This extension will ensure that we continue to have medical evaluation criteria in the listings to adjudicate claims for disability based on impairments in the respiratory body system at step three of our sequential evaluation process. PMID- 11503643 TI - Gastroenterology and urology devices; effective date of requirement for premarket approval of the implanted mechanical/hydraulic urinary continence device. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to require the filing of a premarket approval application (PMA) or a notice of completion of a product development protocol (PDP) for the implanted mechanical/hydraulic urinary continence device, a generic type of medical device intended for the treatment of urinary incontinence. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990 (the SMDA), and the Food and Drug Administration Modernization Act of 1997. PMID- 11503644 TI - Civilian Health and Medical Program of the Uniformed Service (CHAMPUS); prosthetic devices. Office of the Secretary, DoD. Final rule. AB - This final rule implements Section 702 of the National Defense Authorization Act for Fiscal Year 1998, which authorizes purchase of prosthetic devices, as determined by the Secretary of Defense, to be necessary because of significant conditions resulting from trauma, congenital anomalies, or disease. The Act changes the existing limited provisions for prosthetic devices, expanding coverage to include the cost sharing of other prostheses, e.g., noses, ears and fingers. PMID- 11503645 TI - Medicare+Choice: where to from here? PMID- 11503646 TI - Regulations on statements made for dietary supplements concerning the effect of the product on the structure or function of the body; partial stay of compliance. Food and Drug Administration, HHS. Final rule; partial stay of compliance. AB - The Food and Drug Administration (FDA) is announcing a partial stay of compliance for the final rule defining the types of statements that can be made concerning the effect of a dietary supplement on the structure or function of the body for certain dietary supplement products. Dietary supplement products that were labeled, or for which labeling had been printed, on or before January 6, 2000, the publication date of the final rule, are eligible for the stay. This action is in response to two petitions for stay and reconsideration. PMID- 11503647 TI - On the alert. PMID- 11503648 TI - SCHIP in the formative years: an update. PMID- 11503649 TI - Reconstructive pelvic surgery. PMID- 11503650 TI - Psychiatry. PMID- 11503651 TI - Catheter thrombectomy for peripheral arterial clot removal. PMID- 11503652 TI - Food labeling: health claims and labeling statements; dietary fiber and cancer; antioxidant vitamins and cancer; omega-3 fatty acids and coronary heart disease; folate and neural tube defects; revocation. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is revoking its regulations codifying the agency's decision not to authorize the use of health claims for four substance disease relationships in the labeling of foods, including dietary supplements: Dietary fiber and cancer, antioxidant vitamins and cancer, omega-3 fatty acids and coronary heart disease, and the claim that 0.8 milligram (mg) of folate in dietary supplement form is more effective in reducing the risk of neural tube defects than a lower amount in conventional food. This action is being taken in response to a decision of the U.S. Court of Appeals for the D.C. Circuit invalidating these regulations and directing FDA to reconsider whether to authorize the four health claims. This action will result in the removal of the regulations but does not constitute FDA authorization of the four claims. FDA is completing its reconsideration of the claims and expects to issue decisions on all four claims by October 10, 2000. PMID- 11503653 TI - What the publisher can teach the patient: intellectual property and privacy in an era of trusted privication. AB - This article begins with a premise that intellectual property and privacy have something significant and yet understated in common: both are about balancing a creator's desire to control a particular set of data with consumers' desires to access and redistribute that data. Both law and technology influence such balancing, making it more or less palatable to use data for particular purposes- whether one is an individual making a copy of a popular song for a friend, or a hospital selling a list of maternity ward patients to a day care service. In the shadow of the Internet's rapid development and concomitant easing of barriers to data sharing, holders of intellectual property are pairing increased legal protection with the technologies of "trusted systems." I describe how these technologies might allow more thorough mass distribution of data, while allowing publishers to retain unprecedented control over their wares. For instance, an e Book seller might charge one price for a read-only copy that could not be printed or forwarded and charge an additional fee for each copy or printout made. Taking up the case of medical privacy, I then suggest that those who worry about the confidentiality of medical records, particularly as they are digitized by recent congressional mandate, might seek to augment comparatively paltry legal protections with trusted systems technologies. For instance, a trusted system could allow a patient to specify how and by whom her records could be used; within limits, she could allow full access to her primary care physician, while allowing only time-limited access to emergency care providers, non-personally identifiable access to medical researchers, and no access at all for marketing purposes. These technologies could allow for new kinds of privacy protection, without sacrificing the legitimate interests of the consumers of medical records. PMID- 11503655 TI - Physical qualification of drivers; medical examination; certificate. Federal Motor Carrier Safety Administration (FMCSA), DOT. Final rule. AB - This document updates and simplifies the medical examination form that is currently used to determine the physical qualification of commercial motor vehicle (CMV) drivers operating in interstate commerce. The FMCSA takes this action in response to numerous requests from medical examiners to update and simplify the medical examination form that is currently used. This action is intended to reduce the incidence of errors on such forms and to provide more uniform medical examinations of CMV drivers engaged in interstate commerce. The current Federal physical qualification standards tested by medical examiners and recorded on the form will not be revised in this rulemaking. PMID- 11503654 TI - Trusted systems and medical records: lowering expectations. PMID- 11503656 TI - Dental products devices; reclassification of endosseous dental implant accessories. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is reclassifying the manually powered drill bits, screwdrivers, countertorque devices, placement and removal tools, laboratory pieces used for fabrication of dental prosthetics, trial abutments, and other manually powered endosseous dental implant accessories from class III to class I. These devices are intended to aid in the placement or removal of endosseous dental implants and abutments, prepare the site for placement of endosseous dental implants or abutments, aid in the fitting of endosseous dental implants or abutments, aid in the fabrication of dental prosthetics, and be used as an accessory with endosseous dental implants when tissue contact will last less than an hour. FDA is also exempting these devices from premarket notification. This reclassification is on the Secretary of Health and Human Services' own initiative based on new information. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976 (the 1976 amendments), the Safe Medical Devices Act of 1990 (the SMDA), and the Food and Drug Administration Modernization Act of 1997 (FDAMA). PMID- 11503657 TI - Medicaid program; home and community-based services. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with comment period expands State flexibility in providing prevocational, educational, and supported employment services under the Medicaid home and community-based services waiver provisions currently found in section 1915(c) of the Social Security Act (the Act); and incorporates the self implementing provisions of section 4743 of the Balanced Budget Act of 1997 that amends section 1915(c)(5) of the Act to delete the requirements that an individual have prior institutionalization in a nursing facility or intermediate care facility for the mentally retarded before becoming eligible for the expanded habilitation services. In addition, we are making a number of technical changes to update or correct the regulations. PMID- 11503658 TI - Keynote paper: reassessing regulatory compliance. PMID- 11503659 TI - Reassessing the law of preemption. PMID- 11503660 TI - Tort law deference to FDA regulation of medical devices. PMID- 11503661 TI - Rewarding regulatory compliance: the pursuit of symmetry in products liability. PMID- 11503662 TI - Regulatory compliance preclusion of tort liability: limiting the dual-track system. PMID- 11503663 TI - Research that matters: bridging the gap between research and practice in thanatology. AB - This article serves as an Introduction to this two-issue special series of Death Studies on the integration of research and practice in thanatology. After discussing the lack of dialogue between researchers and practitioners in the field, the author identifies some common elements of the research and clinical processes in thanatology. These include the central role of pragmatic theory building and the universal human encounter with loss. The author then offers suggestions for enhancing the exchange between researchers and clinicians, including improvements in theory, methodology, and the dissemination of research findings. Lastly, the individual articles in the series are introduced to the reader. PMID- 11503664 TI - Research, clinical practice, and the human experience: putting the pieces together. AB - The work of the clinician and that of the researcher are presented as representing 2 cultures that cannot exist without each other. A third culture is described to reflect the common human experience of death and grief that invariably informs the work of both clinicians and researchers. At different times in our lives we can be both the subjects and the objects of the work in which we are involved. The author's experience as co-principal investigator with the Massachusetts General Hospital/Harvard Child Bereavement Study and the application of qualitative research is used to exemplify these ideas and demonstrate how the cultures can be integrated. PMID- 11503665 TI - Psychotherapy of traumatic grief: a review of evidence for psychotherapeutic treatments. AB - The authors present the concept of Traumatic Grief and then review controlled studies that pertain to its psychotherapeutic treatment. They conclude that it will most likely be a specific therapy for Traumatic Grief that will be proven most effective in systematic studies completed in the future. PMID- 11503666 TI - Searching for meaning in loss: are clinical assumptions correct. AB - Three assumptions guiding research and clinical intervention strategies for people coping with sudden, traumatic loss are that (a) people confronting such losses inevitably search for meaning, (b) over time most are able to find meaning and put the issue aside, and (c) finding meaning is critical for adjustment or healing. We review existing empirical research that addresses these assumptions and present evidence from a study of 124 parents coping with the death of their infant and a study of 93 adults coping with the loss of their spouse or child to a motor vehicle accident. Results of these studies indicate that (a) a significant subset of individuals do not search for meaning and yet appear relatively well-adjusted to their loss; (b) less than half of the respondents in each of these samples report finding any meaning in their loss, even more than a year after the event; and (c) those who find meaning, although better adjusted than those who search but are unable to find meaning, do not put the issue of meaning aside and move on. Rather, they continue to pursue the issue of meaning as fervently as those who search but do not find meaning. Implications for both research and clinical intervention are discussed. PMID- 11503667 TI - Searching for the meaning of meaning: grief therapy and the process of reconstruction. AB - A comprehensive quantitative review of published randomized controlled outcome studies of grief counseling and therapy suggests that such interventions are typically ineffective, and perhaps even deleterious, at least for persons experiencing a normal bereavement. On the other hand, there is some evidence that grief therapy is more beneficial and safer for those who have been traumatically bereaved. Beginning with this sobering appraisal, this article considers the findings of C.G. Davis, C.B. Wortman, D.R. Lehman, and R.C. Silver (this issue) and their implications for a meaning reconstruction approach to grief therapy, arguing that an expanded conception of meaning is necessary to provide a stronger basis for clinical intervention. PMID- 11503668 TI - Medicare program; additional supplier standards. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule establishes additional standards for an entity to qualify as a Medicare supplier for purposes of submitting claims and receiving payment for durable medical equipment, prosthetics, orthotics, and supplies (DMEPOS). These regulations will ensure that suppliers of DMEPOS are qualified to provide the appropriate health care services and will help safeguard the Medicare program and its beneficiaries from any instances of fraudulent or abusive billing practices. PMID- 11503669 TI - Ramblings from the trenches: a clinical perspective on thanatological research. AB - Is thanatological research helpful in clinical practice, or do clinicians feel researchers are from some "other planet"? Written from the perspective of two practicing clinicians, this article explores the problems and potential of research to enhance clinical practice in end-of-life and bereavement care. Using a case example as a starting point, the article highlights important choice points where good research could assist the clinician. It also discusses several domains of theory and research where advances in knowledge are likely to be particularly helpful to caregivers "in the trenches." Finally, some of the barriers to dissemination of new information to practitioners are described, and suggestions for reducing these barriers are discussed. PMID- 11503670 TI - The use of research findings in bereavement programs: a case study. AB - Violent death is a major public health problem in the United States, yet there is no consensus among bereavement researchers and clinicians regarding a "gold standard" of bereavement services to be offered to family survivors. This article has three purposes: (a) to describe the planning, implementation, and results of a theory-based intervention study involving parents bereaved by the violent deaths of their children; (b) to suggest programmatic elements of bereavement services based on some findings from both the bereavement program and the follow-up data obtained from parents up to 5 years postdeath; and (c) to identify future research and theory development needs. The results of the intervention study involving 261 bereaved parents provide helpful insights regarding parent bereavement program preferences as well as changes in parent outcomes. Some of the most relevant findings pertain to variability in distress levels, gender, and causes of deceased children's deaths. Findings suggest that many different types of services are needed to meet parents' needs. The follow up data collected from the parents 1, 2, and 5 years postdeath demonstrate that loss accommodation following violent death bereavement is both lengthy and difficult. Recommendations follow for both bereavement services and research studies. PMID- 11503671 TI - Grief therapy and research as essential tensions: prescriptions for a progressive partnership. AB - Although the articles in this Special Series take an important step toward fostering mutually respectful dialogue between thanatological research and practice, many impediments still stand in the way of the continuation of the conversation. This closing coda therefore offers several prescriptions for sustaining a constructive exchange between the two domains, in the hope that a more artful and empirically informed grief therapy might yet take shape that embodies the unique strengths of each perspective. PMID- 11503672 TI - A survey of professionals' training and experiences in delivering death notifications. AB - A mail survey was conducted of 240 people from different professions that routinely encountered death to assess their previous training and experiences in delivering death notifications. Nearly 40% of these persons had received neither classroom nor experiential training in death notification, although 70% of respondents had performed at least one notification. The causes of death that contributed to notifiers' distress during notification included (a) violent crime, (b) drunk driving crashes, (c) suicide, and (d) the death of a child. Survivor reactions that were the most difficult for notifiers to manage during the notification included (a) attempts to harm self or others (b) physical acting out, and (c) intense anxiety. Notifiers indicated that they most frequently coped with the stresses of notification by (a) spending time with family, (b) talking with coworkers, and (c) spending time alone. The implications of the results and the needs for systematic death notification education were discussed. PMID- 11503673 TI - Scott and White Grief Study--phase 2: toward an adaptive model of grief. AB - The current study was a dual investigation focused both on the pathogenesis of grief responses and on factors associated with personal growth as a bereavement outcome in a heterogeneous sample of 85 mourners. To examine the pathogenesis of grief, the authors tested the ability of several high-risk factors to predict mourners' subsequent emotional intensity on 2 dependent measures: the Grief Experience Inventory and the Hogan Grief Reaction Checklist. Three situational variables (traumatic death, younger age of decedent, and perception of preventability) as well as 2 mourner liabilities (history of mental health treatment and greater number of other losses) were associated with higher subjective grief misery scores. When using personal growth as a positive outcome following bereavement, the authors identified 4 behavioral correlates of adaptive grieving: ability to see some good resulting from the death, having a chance to say goodbye, intrinsic spirituality, and spontaneous positive memories of the decedent. The advantages of an adaptive model of grief for generating treatment implications are discussed. PMID- 11503674 TI - Cryosurgical ablation of unresectable hepatic tumors. PMID- 11503675 TI - Photodynamic therapy for subfoveal choroidal neovascularization. PMID- 11503676 TI - Radiofrequency ablation of unresectable hepatic tumors. PMID- 11503677 TI - Autologous chondrocyte transplantation. PMID- 11503678 TI - Radiofrequency volumetric tissue reduction for the treatment of simple snoring, upper airway resistance syndrome, or obstructive sleep apnea syndrome. PMID- 11503679 TI - Treatment of twin-twin transfusion syndrome (TTTS) with aggressive amnioreduction and/or fetoscopic laser therapy. PMID- 11503680 TI - Medical devices; labeling for menstrual tampon for the "ultra" absorbency. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule that amends its menstrual tampon labeling regulation to add the term "ultra" absorbency for tampons that absorb 15 to 18 grams (g) of fluid with the syngyna test. At present, FDA requires standardized terms to be used for the labeling of a menstrual tampon to indicate its particular absorbency. This rule enables consumers to compare the absorbency of one brand and style of tampon with the absorbency of other brands and styles. FDA is issuing this final rule under the Federal Food, Drug, and Cosmetic Act (the act) to ensure that labeling of menstrual tampons is not misleading. Elsewhere in this issue of the Federal Register, FDA is proposing to change the standardized menstrual tampon term "junior" to "light". PMID- 11503681 TI - Medical devices; exemption from premarket notification; class II devices; triiodothyronine test system. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is publishing an order granting a petition requesting exemption from the premarket notification requirements for the triiodothyronine test system with certain limitations. This rule will exempt from premarket notification the triiodothyronine test system intended for measuring the hormone triiodothyronine in serum and plasma. FDA is publishing this order in accordance with procedures established by the Food and Drug Administration Modernization Action of 1997 (FDAMA). PMID- 11503682 TI - Americans with Disabilities Act (ADA) accessibility guidelines for buildings and facilities; play areas. Architectural and Transportation Barriers Compliance Board. Final rule. AB - The Architectural and Transportation Barriers Compliance Board (Access Board) is issuing final accessibility guidelines to serve as the basis for standards to be adopted by the Department of Justice for new construction and alterations of play areas covered by the Americans with Disabilities Act (ADA). The guidelines include scoping and technical provisions for ground level and elevated play components, accessible routes, ramps and transfer systems, ground surfaces, and soft contained play structures. The guidelines will ensure that newly constructed and altered play areas meet the requirements of the ADA and are readily accessible to and usable by individuals with disabilities. The Department of Justice must adopt the guidelines as standards for them to be enforceable under the ADA. PMID- 11503683 TI - Medicare program; removal of the requirements for the cardiac pacemaker registry. Health Care Financing Administration, HHS. Final rule. AB - This final rule eliminates all requirements and references regarding the Cardiac Pacemaker Registry (the Registry) in our regulations. It conforms to the Food and Drug Adminstration's (FDA) recent final rule that required any physician and any provider of services who requests or receives Medicare payment for the implantation, removal, or replacement of permanent cardiac pacemaker devices and pacemaker leads to submit certain information to the Registry. We used the information to administer Medicare payment for these devices. This rule implements an Act to Repeal An Unnecessary Medical Device Reporting Requirement passed by Congress to eliminate duplicative and unnecessary reporting. PMID- 11503684 TI - Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); TRICARE Dental Program. Office of the Secretary, DoD. Final rule. AB - This final rule revises the comprehensive CHAMPUS regulation pertaining to the Expanded Active Duty Dependents Benefit Plan, or more commonly referred to as the TRICARE Family Member Dental Plan (TFMDP). The TFMDP limited eligibility to eligible dependents of active duty members (under a call or order that does not specify a period of thirty (30) day or less). Concurrent with the timeframe of the publication of the proposed rule, the Defense Authorization Act for Fiscal Year 2000 (Public Law 106-65, sec. 711) was signed into law and its provisions have been incorporated into this final rule. The Act authorized a new plan, titled the TRICARE dental program (TDP), which allows the Secretary of Defense to offer a comprehensive premium based indemnity dental insurance coverage plan to eligible dependents of active duty members (under a call or order that does not specify a period of thirty (30) days or less), eligible dependents of members of the Selected Reserve and Individual Ready Reserve, and eligible members of the Selected Reserve and Individual Ready Reserve. The Act also struck section 1076b (Selected Reserve dental insurance), or Chapter 55 of title 10, United States Code, since the affected population and the authority for that particular dental insurance plan has been incorporated in 10 U.S.C. 1076a. Consistent with the proposed rule and the provisions of the Defense Authorization Act for Fiscal Year 2000, the final rule places the responsibility for TDP enrollment and a large portion of the appeals program on the dental plan contractor; allows the dental plan contractor to bill beneficiaries for plan premiums in certain circumstances; reduces the former TFMDP enrollment period from twenty-four (24) to twelve (12) months; excludes Reserve component members ordered to active duty in support of a contingency operation from the mandatory twelve (12) month enrollment; clarifies dental plan requirements for different beneficiary populations; simplifies enrollment types and exceptions; reduces cost-shares for certain enlisted grades; adds anesthesia as a covered benefit; provides clarification on the Department's use of the Congressional waiver for surviving dependents; incorporates legislative authority for calculating the method by which premiums may be raised and allowing premium reductions for certain enlisted grades; and reduces administrative burden by reducing redundant language, referencing language appearing in other CFR sections and removing language more appropriate to the actual contract. These improvements will provide Uniformed Service members and families with numerous quality of life benefits that will improve participation in the plan, significantly reduce enrollment errors and positively effect utilization of this important dental plan. The proposed rule was titled the "TRICARE Family Member Dental Plan". PMID- 11503685 TI - Health care perceptions and experiences. AB - Survey results suggest that most people have negative attitudes about health maintenance organizations (HMOs), even members of HMOs who are satisfied with their own care. This Issue Brief illustrates how perceptions of HMOs may color peoples' ratings of their own health care. According to new findings from the Center for Studying Health System Change (HSC), differences in ratings between privately insured people in HMOs and other types of insurance are in part attributable to peoples' perceptions of the type of health plan they are in, not the actual type of plan they are covered by. These results, which have implications for efforts to regulate managed care, suggest that reliance on attitudinal surveys alone are likely to provide a somewhat distorted and more negative view of care in HMOs, thereby exaggerating differences in how people assess the care they receive. PMID- 11503686 TI - Amendment to Executive Order 13078, to expand the role of the National Task Force on Employment of Adults with Disabilities to include a focus on youth. PMID- 11503687 TI - Postmarketing studies for approved human drug and licensed biological products; status reports. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is revising the requirements for annual postmarketing status reports for approved human drug and biological products, and is requiring applicants to submit annual status reports for certain postmarketing studies of licensed biological products. This rule describes the types of postmarketing studies covered by these status reports, the information to be included in the reports, and the type of information that FDA would consider appropriate for public disclosure. This action will implement the Food and Drug Administration Modernization Act of 1997 (FDAMA). PMID- 11503688 TI - Medicare program; revisions to payment policies under the physician fee schedule for calendar year 2001. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with comment period makes several changes affecting Medicare Part B payment. The changes include: refinement of resource-based practice expense relative value units (RVUs); the geographic practice cost indices; resource-based malpractice RVUs; critical care RVUs; care plan oversight and physician certification and recertification for home health services; observation care codes; ocular photodynamic therapy and other ophthalmological treatments; electrical bioimpedance; antigen supply; and the implantation of ventricular assist devices. This rule also addresses the comments received on the May 3, 2000 interim final rule on the supplemental survey criteria and makes modifications to the criteria for data submitted in 2001. Based on public comments we are withdrawing our proposals related to the global period for insertion, removal, and replacement of pacemakers and cardioverter defibrillators and low intensity ultrasound. This final rule also discusses or clarifies the payment policy for incomplete medical direction, pulse oximetry services, outpatient therapy supervision, outpatient therapy caps, HCPCS "G" Codes, and the second 5-year refinement of work RVUs for services furnished beginning January 1, 2002. In addition, we are finalizing the calendar year (CY) 2000 interim physician work RVUs and are issuing interim RVUs for new and revised codes for CY 2001. We are making these changes to ensure that our payment systems are updated to reflect changes in medical practice and the relative value of services. This final rule also announces the CY 2001 Medicare physician fee schedule conversion factor under the Medicare Supplementary Medical Insurance (Part B) program as required by section 1848(d) of the Social Security Act. The 2001 Medicare physician fee schedule conversion factor is $38.2581. PMID- 11503689 TI - Computer-enhanced laparoscopic abdominal surgery. PMID- 11503690 TI - Hand surgery. PMID- 11503691 TI - Reasonable charges for medical care or services. Department of Veterans Affairs. Interim final rule. AB - This document amends the Department of Veterans Affairs (VA) medical regulations concerning "reasonable charges" for medical care or services provided or furnished by VA to a veteran: (1) For a nonservice-connected disability for which the veteran is entitled to care (or the payment of expenses of care) under a health plan contract; (2) For a nonservice-connected disability incurred incident to the veteran's employment and covered under a worker's compensation law or plan that provides reimbursement or indemnification for such care and services; or (3) For a nonservice-connected disability incurred as a result of a motor vehicle accident in a State that requires automobile accident reparations insurance. This document amends the regulations to update databases and other provisions for the purpose of providing more precise charges. PMID- 11503692 TI - Wall Street comes to Washington: market watchers evaluate the health care system. AB - Purchasers are tolerating double-digit premium increases for health plans and agreeing to employee demands for open-access products and wide choice of doctors largely because of the robust economy and tight labor markets, according to a panel of analysts at the fifth annual Wall Street roundtable convened by the Center for Studying Health System Change (HSC). The analysts, who specialize in health care companies, also discussed the challenges facing managed care and how health plans are evolving. This Issue Brief reports on these and other trends, including employers considering defined contribution approaches, techniques to control pharmaceutical spending and the impact of the Internet on the health care system. PMID- 11503693 TI - Are defined contributions a new direction for employer-sponsored coverage? AB - Defined contributions for health benefits are being promoted as the new silver bullet for employers to combat the rising costs of health care, the managed care backlash and the changing climate for employer liability. As interest in this concept grows, so does the number of proposed alternatives for implementing it. Originally called fixed contributions, defined contributions now also refer to cash transfers or vouchers, with reliance on the individual market for health insurance. A more recent angle for defined contributions is using the Internet as an on-line marketplace for purchasing health insurance. This Issue Brief examines defined-contribution strategies and assesses issues relevant to employers, employees and public policy makers. PMID- 11503694 TI - Sickness benefits. Railroad Retirement Board. Final rule. AB - The Railroad Retirement Board (Board) hereby amends its regulations under the Railroad Unemployment Insurance Act (RUIA) to permit a "nurse practitioner" to execute a statement of sickness in support of payments of sickness benefits under the RUIA. The Board does not currently accept statements executed by a nurse practitioner, which in some cases may delay payment of benefits. PMID- 11503695 TI - Traineeships. National Institutes of Health, HHS. Final rule. AB - The National Institutes of Health (NIH) is amending the regulations governing traineeships to add conditions under which NIH may terminate traineeship awards and revise the authorities for the awards. PMID- 11503696 TI - Biological products: reporting of biological product deviations in manufacturing. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is amending the regulation requiring licensed manufacturers of biological products to report errors and accidents in manufacturing that may affect the safety, purity, or potency of a product. FDA also is amending the current good manufacturing practice (CGMP) regulations for blood and blood components to require establishments involved in the manufacture of blood and blood components, including licensed manufacturers, unlicensed registered establishments and transfusion services, to report biological product deviations in manufacturing. The final rule requires licensed manufacturers, unlicensed registered blood establishments, and transfusion services who had control over the product when a deviation occurred to report to FDA the biological product deviation if the product has been distributed. The final rule also establishes a 45-day reporting period. FDA is issuing the final rule as part of a retrospective review under Executive Order 12866 of significant FDA regulations to improve the effectiveness of FDA's regulatory program. PMID- 11503697 TI - Exemption from federal preemption of state and local cigarette and smokeless tobacco requirements; revocation. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is revoking its regulation governing the exemption from Federal preemption of State and local medical device requirements for the sale and distribution of cigarettes and smokeless tobacco to children and adolescents. This action is being taken in response to the Supreme Court Decision of March 21, 2000, in which the court held that Congress has not given FDA the authority to regulate tobacco products as customarily marketed. On March 31, 2000, FDA removed its regulations restricting the sale and distribution of cigarettes and smokeless tobacco to children and adolescents. Because these regulations are not in effect, the State requirements are not preempted. Therefore, FDA is revoking its regulations exempting the State and local requirements from preemption. This rule is also adding a regulation that was inadvertently removed in a previous document. PMID- 11503698 TI - VA payment for non-VA public or private hospital care and non-VA physician services that are associated with either outpatient or inpatient care. Department of Veterans Affairs. Final rule. AB - This document amends our medical regulations concerning VA payment for non-VA public or private hospital care provided to eligible VA beneficiaries. This document also amends our medical regulations concerning VA payment for non-VA physician services that are associated with either outpatient or inpatient care provided to eligible VA beneficiaries at non-VA facilities. With certain exceptions, these payments have been based on Medicare methodology. Sometimes VA can negotiate contracts with hospitals or physicians or with their agents to reduce the payment amounts. This document amends these regulations to allow VA to make lower payments based on such negotiations. PMID- 11503699 TI - Living well: a diagnosis of America's healthiest places. PMID- 11503701 TI - The lifestyle Rx. PMID- 11503700 TI - How to beat the flu: the latest options as a vaccine shortage looms. PMID- 11503702 TI - The U.S. spends more money on health care than any other country. But the nation's 44 million uninsured face a system of . . . second-class medicine. PMID- 11503703 TI - Herbal Rx for prostate problems. PMID- 11503704 TI - How does managed care do it? AB - Integrating the health services and insurance industries, as health maintenance organizations (HMOs) do, could lower expenditure by reducing either the quantity of services or unit price or both. We compare the treatment of heart disease in HMOs and traditional insurance plans using two datasets from Massachusetts. The nature of these health problems should minimize selection. HMOs have 30% to 40% lower expenditures than traditional plans. Both actual treatments and health outcomes differ little; virtually all the difference in spending comes from lower unit prices. Managed care may yield substantial increases in measured productivity relative to traditional insurance. PMID- 11503705 TI - Looking straight ahead. PMID- 11503706 TI - Other causes of choroidal neovascularization. PMID- 11503707 TI - Medicare program; prospective payment system for hospital outpatient services. Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This interim final rule with comment period provides for the annual update to the Medicare hospital outpatient prospective payment system conversion factor that is used to calculate the payment amount for each payment group, effective January 1, 2001. It also updates the wage index values and incorporates the year 2001 changes in the procedure codes that are used to make payments under this system. In this rule, we are also responding to public comments received on those portions of the April 7, 2000 final rule with comment period (which established the hospital outpatient prospective payment system) that implemented related provisions of the Balanced Budget Refinement Act (BBRA) of 1999. In addition, we are responding to public comments on the August 3, 2000 interim final rule with comment period that modified the April 7, 2000 final rule with comment period by revising the criteria used to define new or innovative medical devices, drugs, and biologicals eligible for transitional pass-through payments and correcting the criteria for grandfathering provider-based Federally Qualified Health Centers (FQHC) into the prospective payment system. PMID- 11503708 TI - Asbestos worker protection. Environmental Protection Agency (EPA). Final rule. AB - In this Final Rule, EPA is amending both the Asbestos Worker Protection Rule (WPR) and the Asbestos-in-Schools Rule. The WPR amendment protects State and local government employees from the health risks of exposure to asbestos to the same extent as private sector workers by adopting for these employees the Asbestos Standards of the Occupational Safety and Health Administration (OSHA). The WPR's coverage is extended to State and local government employees who are performing construction work, custodial work, and automotive brake and clutch repair work. This final rule cross-references the OSHA Asbestos Standards for Construction and for General Industry, so that future amendments to these OSHA standards are directly and equally effective for employees covered by the WPR. EPA also amends the Asbestos-in-Schools Rule to provide coverage under the WPR for employees of public local education agencies who perform operations, maintenance, and repair activities. EPA is issuing this final rule under section 6 of the Toxic Substances Control Act (TSCA). PMID- 11503709 TI - Small business size standards; health care. Small Business Administration. Final rule. AB - The Small Business Administration is adopting new size standards for 19 Health Care industries and retaining the existing $5 million size standard for the remaining 11 Health Care industries. The North American Industry Classification System classifies Health Care industries under Subsector 621, Ambulatory Health Care Services; Subsector 622, Hospitals; and Subsector 623, Nursing and Residential Care Facilities. These revisions are made to more appropriately define the size of businesses in these industries that SBA believes should be eligible for Federal small business assistance programs. PMID- 11503710 TI - Amendments to Summary Plan Description regulations. Pension and Welfare Benefits Administration, Labor. Final rule. AB - This document contains a final rule amending the regulations governing the content of the Summary Plan Description (SPD) required to be furnished to employee benefit plan participants and beneficiaries under the Employee Retirement Income Security Act of 1974, as amended (ERISA). These amendments implement information disclosure recommendations of the President's Advisory Commission on Consumer Protection and Quality in the Health Care Industry, as set forth in their November 20, 1997, report, "Consumer Bill of Rights and Responsibilities." Specifically, the amendments clarify benefit, medical provider, and other information required to be disclosed in, or as part of, the SPD of a group health plan and repeal the limited exemption with respect to SPDs of welfare plans providing benefits through qualified health maintenance organizations (HMOs). In addition, this document contains several amendments updating and clarifying provisions relating to the content of SPDs that affect both pension and welfare benefit plans. This document also adopts in final form certain regulations that were effective on an interim basis implementing amendments to ERISA enacted as part of the Health Insurance Portability and Accountability Act of 1996 (HIPAA). This final rule will affect employee pension and welfare benefit plans, including group health plans, as well as administrators, fiduciaries, participants and beneficiaries of such plans. PMID- 11503711 TI - Employee Retirement Income Security Act of 1974: rules and regulations for administration and enforcement; claims procedure. Pension and Welfare Benefits Administration, Labor. Final regulation. AB - This document contains a final regulation revising the minimum requirements for benefit claims procedures of employee benefit plans covered by Title I of the Employee Retirement Income Security Act of 1974 (ERISA or the Act). The regulation establishes new standards for the processing of claims under group health plans and plans providing disability benefits and further clarifies existing standards for all other employee benefit plans. The new standards are intended to ensure more timely benefit determinations, to improve access to information on which a benefit determination is made, and to assure that participants and beneficiaries will be afforded a full and fair review of denied claims. When effective, the regulation will affect participants and beneficiaries of employee benefit plans, employers who sponsor employee benefit plans, plan fiduciaries, and others who assist in the provision of plan benefits, such as third-party benefits administrators and health service providers or health maintenance organizations that provide benefits to participants and beneficiaries of employee benefit plans. PMID- 11503712 TI - Senior living: beyond the nursing home. PMID- 11503713 TI - Immunology and Microbiology Devices; Classification of Anti-Saccharomyces cerevisiae (S. cerevisiae) Antibody (ASCA) Test Systems. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the Anti-Saccharomyces cerevisiae (S. cerevisiae) antibody (ASCA) test system into class II (special controls). The special control that will apply to this device is a guidance document entitled "Guidance for Industry and FDA Reviewers: Class II Special Control Guidance Document for Anti-Saccharomyces cerevisiae (S. cerevisiae) Antibody (ASCA) Premarket Notifications." Elsewhere in this issue of the Federal Register. FDA is announcing the availability of this guidance document. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying these devices into class II (special controls) in order to provide a reasonable assurance of the safety and effectiveness of the devices. PMID- 11503714 TI - Risk management. National Aeronautics and Space Administration (NASA). Interim rule adopted as final with changes. AB - This is a final rule amending the NASA FAR Supplement (NFS) to emphasize considerations of risk management, including safety, security (including information technology security), health, export control, and damage to the environment, within the acquisition process. This final rule addresses risk management within the context of acquisition planning, selecting sources, choosing contract type, structuring award fee incentives, administering contracts, and conducting contractor surveillance. PMID- 11503715 TI - Conscious sedation. PMID- 11503716 TI - Anesthesiology. PMID- 11503717 TI - Exploration of the relationship between physical and/or sexual abuse, attitudes about life and death, and suicidal ideation in young women. AB - Experiencing sexual and/or physical abuse at any point in one's life can be quite traumatic. In addition to the immediate effects of the abuse, numerous long-term consequences have been identified, including an increased risk of developing psychological disorders and attempting suicide. We sought to determine if a relationship exists between abuse experiences, attitudes about life and death, and suicidal ideation by surveying 644 college women between the ages of 18 and 24. Participants completed packets containing standardized abuse questionnaires, the Multi-Attitude Suicide Tendency Scale, and the Adult Suicidal Ideation Questionnaire. We concluded that abuse plays a role in how these young women think about life and death. Specifically, less repulsion by death and more repulsion by life were reported by women with more significant abuse histories. In addition, these same women endorsed higher levels of current suicidal ideation. It was also determined that women who had experienced more abuse tended to have been in therapy for longer periods of time. Implications of this study for mental health professionals working with suicidal young adults are discussed. PMID- 11503718 TI - Traumatic stress and death anxiety among community residents exposed to an aircraft crash. AB - This article examined the relationship between traumatic stress and death anxiety among community residents who lived near woodlands in which an aircraft had crashed in Coventry, England. The hypothesis was that there would be a high level of impact of the crash experienced by residents and that the residents would experience psychological distress. It was also hypothesized that the impact of the crash and distress were associated with death anxiety. Eighty-two residents were interviewed for the study. They were asked to complete the Impact of Event Scale, the General Health Questionnaire (GHQ), and the Death Anxiety Scale. The results showed that Coventry residents were found to experience intrusive thoughts and display avoidance behavior. The residents' intrusive thoughts and avoidance behavior were significantly more severe than one group of Horowitz's standardized samples (i.e., the medical students), but not significantly different from another group (i.e., the stress clinic samples). Fifty-seven percent scored at or above the GHQ cutoff point, which meant that they were considered to be psychiatric cases. Community residents scored significantly lower in death anxiety than the standardized high-death anxiety patients but no differently from the standardized control patients. Correlations were found between the impact of the event, psychological distress, and death anxiety. PMID- 11503719 TI - Firearms and suicide: the American experience, 1926-1996. AB - The hypothesis that American male suicide rates are higher than the rates of women because men select more highly lethal methods than do women is tested by adjusting male rates so that the distribution of male and female suicides to highly lethal methods is equal. However, the adjusted male rate is still higher than the total female rate in all eight periods from 1926-1929 through 1996. Also, increases in the percentage of female suicides using firearms over this period are unrelated to increases in female rates, and similar increases in firearms use by males are positively related to increases in male suicide rates only in recent decades. The impact of change in the male firearms suicide rate on change in their total suicide rate was weak or nonexistent in three of seven change periods; its impact on the female total rate was trivial in five of the seven change periods. PMID- 11503720 TI - Factor structure of the Grief Experience Questionnaire (GEQ). AB - The Grief Experience Questionnaire (GEQ; T.W. Barrett & T.B. Scott, 1989) is a self-report measure of grief responses, including some that have been associated with grief after a suicide (e.g., feelings of rejection, responsibility, shame, stigmatization, etc.). In this study, a sample of 350 university students who had experienced the death of a significant other completed the GEQ. A principal components analysis with varimax rotation yielded an 8-factor solution with satisfactory psychometric properties. Results clearly document that the GEQ is a multidimensional measure of grief phenomenology. It is concluded that although the GEQ has broad applicability, the scale as revised herein may have a special relevance to suicide bereavement, and may be of use in both research and clinically based applications. PMID- 11503721 TI - Developing beliefs that are compatible with death: revising our assumptions about predictability, control, and continuity. AB - Many of us assume that life is highly predictable, subject to our control, and destined to continue indefinitely. Although such perspectives on life afford us a sense of security, these views tend to be incompatible with the experience of death. This article outlines the benefits of assimilating death into our belief systems and explores problems with waiting to alter our beliefs until after confrontation with death. Next, the article describes how we can use the strategy of imagining our own death or the death of loved ones as a means of making our beliefs more compatible with death. Options are then presented for overcoming resistances to imagining loss. Finally, the article proposes alternate ways of conceptualizing issues of predictability, control, and continuity; it addresses procedures for sustaining new beliefs in these areas; and it lists limitations associated with attempting to adopt such beliefs. PMID- 11503722 TI - The federal-state Medicaid match: an ongoing tug-of-war over practice and policy. PMID- 11503723 TI - Food labeling, safe handling statements, labeling of shell eggs; refrigeration of shell eggs held for retail distribution. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is revising its food labeling regulations to require a safe handling statement on cartons of shell eggs that have not been treated to destroy Salmonella microorganisms. The agency also is requiring that, when held at retail establishments, shell eggs be stored and displayed under refrigeration at a temperature of 7.2 degrees C (45 degrees F) or less. FDA is taking these actions because of the number of outbreaks of foodborne illnesses and deaths caused by Salmonella Enteritidis (SE) that are associated with the consumption of shell eggs. These actions also respond, in part, to petitions from Rose Acres Farm, Inc., and the Center for Science in the Public Interest (CSPI). Safe handling statements will help consumers take measures to protect themselves from illness or deaths associated with consumption of shell eggs that have not been treated to destroy Salmonella (all serotypes). Refrigeration of shell eggs that have not been treated to destroy Salmonella will help prevent the growth of SE in shell eggs. PMID- 11503724 TI - Medical device; exemption from premarket notification; class II devices; barium enema retention catheters and tips with or without a bag. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is publishing an order granting a petition requesting exemption from the premarket notification requirements for barium enema retention catheters and tips with or without a bag with certain limitations. This rule will exempt from premarket notification barium enema retention catheters and tips with or without a bag. FDA is publishing this order in accordance with procedures established by the Food and Drug Administration Modernization Act of 1997 (FDAMA). PMID- 11503725 TI - Special demonstration programs. Office of Special Education and Rehabilitative Services, Department of Education. Final regulations. AB - The Secretary issues regulations governing the Special Demonstration Programs. These regulations are needed to implement changes in the Rehabilitation Act Amendments of 1998. The regulations provide definitions and requirements for grants and contracts under the expanded authority of the Special Demonstration Programs. PMID- 11503726 TI - Revision to requirements for licensed Anti-Human Globulin and Blood Grouping Reagents. Food and Drug Administration, HHS. Direct final rule. AB - The Food and Drug Administration (FDA) is amending the biologics regulations applicable to microbiological controls for licensed Anti-Human Globulin (AHG) and Blood Grouping Reagents (BGR). FDA is amending the regulations to remove the requirements that the products be sterile. FDA is publishing this direct final rule because the requirement that these products be sterile is not necessary for the products to be safe, pure, and potent. FDA is issuing these amendments directly as a final rule because they are noncontroversial and there is little likelihood that FDA will receive any significant comments opposing the rule. Elsewhere in this issue of the Federal Register, FDA is publishing a proposed rule under FDA's usual procedures for notice and comment in the event the agency receives any significant adverse comments. If FDA receives any significant adverse comment that warrants terminating the direct final rule, FDA will consider such comments on the proposed rule in developing the final rule. PMID- 11503727 TI - Prevalence of urinary incontinence in middle-aged and older women: a survey-based methodological experiment. AB - OBJECTIVE: To determine whether self-reports of urinary incontinence are influenced by the context in which survey questions are presented. Variation in the extent to which surveys promote complete and accurate reporting may help explain the wide range of published prevalence estimates. METHODS: Respondents to a nationally representative telephone survey were randomly assigned one of two questionnaire forms. Both forms contained the same incontinence question. One form included an introduction and follow-up probe, which acknowledged the embarrassment of discussing incontinence and stressed the importance of thorough reporting. RESULTS: Use of the introduction and probe produced a significantly higher prevalence rate, with the effect varying by respondent's age. DISCUSSION: The prevalence of incontinence is likely to be underestimated unless studies attend to the possibility of biased reporting. The finding of a form by age interaction suggests the importance of survey context in studies of risk factors, as well as in prevalence studies. PMID- 11503728 TI - Sustained personal autonomy: a measure of successful aging. AB - OBJECTIVES: This study addresses the following question: What characteristics of urban, noninstitutionalized elders predict which individuals are most likely to remain independent of personal assistance during a 2-year observation period? METHODS: A population-based sample of 602 noninstitutionalized urban residents aged 70 and older was followed for 2 years. RESULTS: Ninety-eight of the 487 survivors remained independent. Factors associated with sustained independence were relatively younger age, male gender, fewer medical conditions, good physical function, and nonsmoking. The attitudes "favors family or self over agency assistance" and "does not expect filial obligation" were also independently associated. DISCUSSION: The results are consistent with previous studies of successful aging and show that attitudes expressed at baseline favoring personal independence are associated with sustained autonomy during a period of at least 2 years. PMID- 11503729 TI - The cost of recruiting Alzheimer's disease caregivers for research. AB - OBJECTIVES: This study examined the cost and outcomes associated with multiple recruitment strategies used to enroll participants in an Alzheimer's disease (AD) caregiver study. METHODS: Recruitment data were collected as part of an AD caregiving intervention study and examined for number of referrals and participants, yield, personnel cost, materials cost, total cost, and cost per participant. RESULTS: Recruitment rates varied by method implemented and referral source. Overall, 100 participants were enrolled for a total cost of $10,127, yielding an average per participant cost of $101.00. The Formal Recruitment Method, used at agencies with large participant pools, emerged as the most cost effective strategy. DISCUSSION: Recruitment costs varied greatly, depending on the referral sites and their available pool of participants. Substantial time, money, and personnel need to be budgeted for recruitment efforts given the expected costs, the competing pressures in the health care region, and the variability of participant response. PMID- 11503730 TI - Age-based rationing in the allocation of health care. AB - OBJECTIVES: This article seeks to review debates about age-based rationing in health care. METHODS: The article identifies four different levels (or types) of decision-making in health resources allocation--societal, strategic, programmatic, and clinical--and assesses how the issues of rationing vary in relation to each level. RESULTS: The article concludes that rationing is least defensible at the clinical level, where it is also most covert. The role of rationing at other levels is more defensible when based on grounds of cost effectiveness rather than equity. The article emphasizes the importance of fairness in health allocation and suggests that efficiency criteria need to be considered in that context. DISCUSSION: The article suggests that rationing is most problematic where it is least overt. This raises further questions about how rationing can be made more explicit at different levels of decision making. PMID- 11503731 TI - Job characteristics and leisure physical activity. AB - OBJECTIVES: This study employs a sample population of older workers to estimate an empirical model of leisure exercise activity. Alternative theories relating work and leisure attitudes relevant for understanding the exercise behavior of older workers are tested empirically. METHODS: Responses of 6,433 full-time older workers (51 to 61 years old) from the 1992 Health and Retirement Study (HRS) are grouped into two white-collar and blue-collar worker categories and are analyzed to test whether self-reported levels of regular physical activity are associated with the physical demands and stress associated with one's job. RESULTS: Although the white-collar workers, whose jobs involve more physical efforts, are more likely to do light physical activity, the blue-collar workers, whose jobs are more physically demanding, tend to engage in more vigorous exercise. DISCUSSION: The empirical results are most supportive of the generalization theory, and they also illustrate the complexity of relationships between work and leisure physical activity. PMID- 11503732 TI - Use of mental health services by persons with a mental illness in nursing facilities: initial impacts of OBRA87. AB - OBJECTIVE: To identify mental health service use patterns in nursing facilities subsequent to nursing home reforms in the Omnibus Budget Reconciliation Act of 1987. METHODS: The study involves a retrospective analysis of Medicare Part B claims for mental health care for persons in a nursing facility in 1992 (N = 692) using the Medicare Current Beneficiary Survey. Mental health service use was described and analyzed using logistic regression. RESULTS: Only 26% of nursing facility residents and 36% of residents with a mental illness had a mental health visit in 1992. Logistic regression confirms analyses prior to 1987 showing older residents and those in rural areas remain less likely to receive mental health visits. DISCUSSION: Claims data collected after the passage of The Nursing Home Reform Act (NHRA) of 1987 indicate continued low levels of mental health treatment in nursing homes. PMID- 11503733 TI - Procedures for transportation workplace drug and alcohol testing programs. Office of the Secretary, DOT. Final rule. AB - The Department of Transportation is revising its drug and alcohol testing procedures regulation. The purposes of the revision are to make the organization and language of the regulation clearer, to incorporate guidance and interpretations of the rule into its text, and to update the rule to include new provisions responding to changes in technology, the testing industry, and the Department's program. PMID- 11503734 TI - Regulations implementing the Federal Coal Mine Health and Safety Act of 1969, as amended. Employment Standards Administration, Labor. Final rule. AB - On January 22, 1997, the Department issued a proposed rule to amend the regulations implementing the Black Lung Benefits Act. 62 FR 3338-3435 (Jan. 22, 1997). When the comment period closed on August 21, 1997, the Department had received written submissions from almost 200 interested persons, including coal miners, coal mine operators, insurers, physicians, and attorneys. The Department also held hearings in Charleston, West Virginia, and Washington, D.C. at which over 50 people testified. The Department carefully reviewed the testimony and the comments and, on October 8, 1999, issued a second notice of proposed rulemaking. 64 FR 54966-55072 (Oct. 8, 1999). In its second notice, the Department proposed changing several of the most important provisions in its initial proposal. The Department also explained its decision not to alter the original proposal with respect to other key regulations based on the comments received to date. Finally, the Department prepared an initial regulatory flexibility analysis. In order to ensure that small businesses that could be affected by the Department's proposal received appropriate notice of the Department's proposed changes, the Department mailed a copy of the second notice of proposed rulemaking to all coal mine operators contained in the databases maintained by the Mine Safety and Health Administration. The Department initially allowed interested parties until December 7, 1999 to file comments to its second proposal, but extended that period until January 6, 2000. The Department received 37 written submissions before the close of the comment period, from groups representing both coal miners and coal mine operators. The Department also received comments from individual miners, various coal mining and insurance companies, as well as from claims processing organizations, attorneys, and various professional organizations. The Department has carefully reviewed all of the comments, and is issuing its final rule. The rule contains a final regulatory flexibility analysis as required by the Regulatory Flexibility Act. PMID- 11503735 TI - Code of conduct for the International Space Station Crew. National Aeronautics and Space Administration (NASA). Interim final rule. AB - NASA is issuing new regulations entitled "International Space Station Crew," to implement certain provisions of the International Space Station (ISS) Intergovernmental Agreement (IGA) regarding ISS crewmembers' observance of an ISS Code of Conduct. PMID- 11503736 TI - National Medical Support Notice. Pension and Welfare Benefits Administration, Labor. Final rule. AB - This document contains a final rule that promulgates a National Medical Support Notice to be issued by State agencies as a means of enforcing the health care coverage provisions in a child support order, and to be treated by plan administrators of group health plans as a qualified medical child support order under section 609(a) of Title I of the Employee Retirement Income Security Act (ERISA). Through this regulation, the Department of Labor (the Department) is implementing an amendment to section 609(a) of ERISA, made by section 401 of the Child Support Performance and Incentive Act of 1998 (CSPIA), Pub. L. 105-200. This rule will affect group health plans, participants in group health plans, noncustodial children of such participants, and State agencies that administer child support enforcement programs. PMID- 11503737 TI - National Medical Support Notice. Office of Child Support Enforcement, Administration for Children and Families, HHS. Final rule. AB - This rule implements provisions of the Child Support Performance and Incentives Act of 1998 (CSPIA), Public Law 105-200, that require State child support enforcement agencies, under title IV-D of the Social Security Act (the Act), to enforce the health care coverage provision in a child support order through the use of the National Medical Support Notice (NMSN). A proposed rule was published in the Federal Register on November 15, 1999 (64 FR 62074). After consideration of the written comments received, changes have been made in this final regulation, including changes to the NMSN found in the Appendix. PMID- 11503738 TI - Standards for privacy of individually identifiable health information. Office of the Assistant Secretary for Planning and Evaluation, DHHS. Final rule. AB - This rule includes standards to protect the privacy of individually identifiable health information. The rules below, which apply to health plans, health care clearinghouses, and certain health care providers, present standards with respect to the rights of individuals who are the subjects of this information, procedures for the exercise of those rights, and the authorized and required uses and disclosures of this information. The use of these standards will improve the efficiency and effectiveness of public and private health programs and health care services by providing enhanced protections for individually identifiable health information. These protections will begin to address growing public concerns that advances in electronic technology and evolution in the health care industry are resulting, or may result in, a substantial erosion of the privacy surrounding individually identifiable health information maintained by health care providers, health plans and their administrative contractors. This rule implements the privacy requirements of the Administrative Simplification subtitle of the Health Insurance Portability and Accountability Act of 1996. PMID- 11503739 TI - Old-age, survivors, and disability insurance and Supplemental Security Income for the aged, blind, and disabled; substantial gainful activity amounts; "services" for trial work period purposes--monthly amounts; student child earned income exclusion. Social Security Administration. Final rules. AB - We are revising the rules to automatically adjust each year, based on any increases in the national average wage index, the average monthly earnings guideline we use to determine whether work done by persons with impairments other than blindness is substantial gainful activity; provide that we will ordinarily find that an employee whose average monthly earnings are not greater than the "primary substantial gainful activity amount," has not engaged in substantial gainful activity without considering other information beyond the employee's earnings; increase the minimum amount of monthly earnings and the minimum number of self-employed work hours in month that we consider shows that a person receiving title II Social Security benefits based on disability is performing or has performed "services" during a trial work period, and automatically adjust the earnings amount each year thereafter; increase the maximum monthly and yearly Student Earned Income Exclusion amounts we use in determining Supplemental Security Income (SSI) Program eligibility and payment amounts for student children, and automatically adjust the monthly and yearly exclusion amounts each year thereafter. We are revising these rules as part of our efforts to encourage individuals with disabilities to test their ability to work and keep working. We expect that these changes will provide greater incentives for many beneficiaries to attempt to work or, if already working, to continue to work or increase their work effort. PMID- 11503740 TI - Medicare, Medicaid, and CLIA programs; extension of certain effective dates for clinical laboratory requirements under CLIA. Centers for Disease Control and Prevention (CDC) and Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule extends certain effective dates for clinical laboratory requirements in regulations published on February 28, 1992, that implemented provisions of the Clinical Laboratory Improvement Amendments of 1988 (CLIA). This rule extends the phase-in date of the quality control requirements applicable to moderate and high complexity tests and extends the date by which an individual with a doctoral degree must possess board certification to qualify as a director of a laboratory that performs high complexity testing. These effective dates are extended to allow the Department to revise quality control requirements and establish the qualification requirements necessary for individuals with doctoral degrees to serve as directors of laboratories performing high complexity testing. These effective date extensions do not reduce the current requirements for quality test performance. PMID- 11503741 TI - Medicare program; expanded coverage for outpatient diabetes self-management training and diabetes outcome measurements. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule implements section 4105 of the Balanced Budget Act of 1997 (BBA) by expanding Medicare coverage for outpatient diabetes self-management training and establishes outcome measurements for evaluating the improvement of the health status of Medicare beneficiaries with diabetes. These services include education and training furnished to a beneficiary with diabetes by an approved entity deemed to meet certain quality standards established in this final rule. The physician (or qualified nonphysician practitioner) treating the beneficiary's diabetes must certify that these services are needed as part of the beneficiary's comprehensive plan of care. PMID- 11503742 TI - At the brink: how Harvard Pilgrim got in trouble. AB - The Massachusetts insurance commissioner placed Harvard Pilgrim Health Care (HPHC) in receivership in January on the basis of large projected losses that put the nonprofit plan in a significant negative net worth position. Because Harvard Pilgrim was the largest health plan in the market, with substantial amounts payable to hospitals and physicians, its financial problems shook the Boston health care community. The story also attracted national attention because of the plan's prominence and its reputation for quality. The Center for Studying Health System Change (HSC) followed the Harvard Pilgrim story closely as part of its continuous tracking of Boston--one of the 12 Community Tracking Study sites visited every two years--and is able to put this event in broader context. Many of Harvard Pilgrim's problems are evident in plans elsewhere. This Issue Brief discusses the causes of the plan's financial problem and the state's response, which has preserved the organization. PMID- 11503743 TI - Pathology. PMID- 11503744 TI - Harmonic scalpel tonsillectomy. PMID- 11503746 TI - Is assisted living the right choice? PMID- 11503745 TI - Medicare and Medicaid programs; physicians' referrals to health care entities with which they have financial relationships. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with 90-day comment period (Phase I of this rulemaking) incorporates into regulations the provisions in paragraphs (a), (b), and (h) of section 1877 of the Social Security Act (the Act). Under section 1877, if a physician or a member of a physician's immediate family has a financial relationship with a health care entity, the physician may not make referrals to that entity for the furnishing of designated health services (DHS) under the Medicare program, unless an exception applies. The following services are DHS: clinical laboratory services; physical therapy services; occupational therapy services; radiology services, including magnetic resonance imaging, computerized axial tomography scans, and ultrasound services; radiation therapy services and supplies; durable medical equipment and supplies; parenteral and enteral nutrients, equipment, and supplies; prosthetics, orthotics, and prosthetic devices and supplies; home health services; outpatient prescription drugs; and inpatient and outpatient hospital services. In addition, section 1877 of the Act provides that an entity may not present or cause to be presented a Medicare claim or bill to any individual, third party payer, or other entity for DHS furnished under a prohibited referral, nor may we make payment for a designated health service furnished under a prohibited referral. Paragraph (a) of section 1877 of the Act includes the general prohibition. Paragraph (b) of the Act includes exceptions that pertain to both ownership and compensation relationships, including an in-office ancillary services exception. Paragraph (h) includes definitions that are used throughout section 1877 of the Act, including the group practice definition and the definitions for each of the DHS. We intend to publish a second final rule with comment period (Phase II of this rulemaking) shortly addressing, to the extent necessary, the remaining sections of the Act. Phase II of this rulemaking will address comments concerning the ownership and investment exceptions in paragraphs (c) and (d) and the compensation exceptions in paragraph (e) of section 1877 of the Act. Phase II of this rulemaking will also address comments concerning the reporting requirements and sanctions provided by paragraphs (f) and (g) of the Act, respectively, and include further consideration of the general exception to the referral prohibition related to both ownership/investment and compensation for services furnished in an ambulatory surgical center (ASC), end-stage renal dialysis facility, or by a hospice in section 411.355(d) of the regulations (this exception presently is in force and effect as to clinical laboratory services). In addition, Phase II of this rulemaking will address section 1903(s) of the Act, which extends aspects of the referral prohibition to the Medicaid Program. Phase II will also address comments received in response to this rulemaking, as appropriate, and certain proposals for new exceptions to section 1877 of the Act not included in the 1998 proposed rulemaking, but suggested in the public comments. PMID- 11503747 TI - Super-germ alert: how to avoid antibiotic misuse and overuse. PMID- 11503748 TI - Engineering genetics policy. PMID- 11503749 TI - Quackery no more: alternative medicine moves into the mainstream. PMID- 11503750 TI - Futurespeak. Interview by Alison Stein Wellner. PMID- 11503751 TI - Interim final rules for nondiscrimination in health coverage in the group market. Internal Revenue Service, Department of the Treasury; Pension and Welfare Benefits Administration, Department of Labor; Health Care Financing Administration, Department of Health and Human Services. Interim final rules with request for comments. AB - This document contains interim final rules governing the provisions prohibiting discrimination based on a health factor for group health plans and issuers of health insurance coverage offered in connection with a group health plan. The rules contained in this document implement changes made to the Internal Revenue Code of 1986 (Code), the Employee Retirement Income Security Act of 1974 (ERISA), and the Public Health Service Act (PHS Act) enacted as part of the Health Insurance Portability and Accountability Act of 1996 (HIPAA). PMID- 11503752 TI - Assistance to States for the Education of Children with Disabilities. Office of Special Education and Rehabilitative Services, Department of Education. Final regulations. AB - The Secretary amends the regulations for the Assistance to States for the Education of Children with Disabilities program under Part B of the Individuals with Disabilities Education Act (IDEA; Part B). This amendment is needed to implement the statutory provision that for any fiscal year in which the appropriation for section 611 of the IDEA exceeds $4.1 billion, a local educational agency (LEA) may treat as local funds up to 20 percent of the amount it receives that exceeds the amount it received during the prior fiscal year. The amendment is intended to ensure effective implementation of the 20 percent rule by clarifying which funds under Part B of IDEA can be included in the 20 percent calculation, and, as a result, to reduce the potential for audit exceptions. PMID- 11503753 TI - When will it change? PMID- 11503754 TI - Revisions to the requirements applicable to blood, blood components, and source plasma; confirmation in part and technical amendment. Food and Drug Administration, HHS. Direct final rule; confirmation in part and technical amendment. AB - The Food and Drug Administration (FDA) is confirming in part the direct final rule issued in the Federal Register of August 19, 1999. The direct final rule amends the biologics regulations by removing, revising, or updating specific regulations applicable to blood, blood components, and Source Plasma to be more consistent with current practices in the blood industry and to remove unnecessary or outdated requirements. FDA is confirming the provisions for which no significant adverse comments were received. The agency received significant adverse comments on certain provisions and is amending Title 21 Code of Federal Regulations to reinstate the former provisions. PMID- 11503755 TI - Medicaid program; revision to Medicaid upper payment limit requirements for hospital services, nursing facility services, intermediate care facility services for the mentally retarded, and clinic services. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule modifies the Medicaid upper payment limits for inpatient hospital services, outpatient hospital services, nursing facility services, intermediate care facility services for the mentally retarded, and clinic services. For each type of Medicaid inpatient service, existing regulations place an upper limit on overall aggregate payments to all facilities and a separate aggregate upper limit on payments made to State-operated facilities. This final rule establishes an aggregate upper limit that applies to payments made to government facilities that are not State government-owned or operated, and a separate aggregate upper limit on payments made to privately-owned and operated facilities. This rule also eliminates the overall aggregate upper limit that had applied to these services. With respect to outpatient hospital and clinic services, this final rule establishes an aggregate upper limit on payments made to State government-owned or operated facilities, an aggregate upper limit on payments made to government facilities that are not State government-owned or operated, and an aggregate upper limit on payments made to privately-owned and operated facilities. These separate upper limits are necessary to ensure State Medicaid payment systems promote economy and efficiency. We are allowing a higher upper limit for payment to non-State public hospitals to recognize the higher costs of inpatient and outpatient services in public hospitals. In addition, to ensure continued beneficiary access to care and the ability of States to adjust to the changes in the upper payment limits, the final rule includes a transition period for States with approved rate enhancement State plan amendments. PMID- 11503756 TI - Medicare program; payment for nursing and allied health education. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule sets forth in regulations Medicare policy for the payment of costs of approved nursing and allied health education programs. In addition, the rule clarifies the payment methodology for certified registered nurse anesthetist education programs. In general, the final rule clarifies and restates payment policies previously established in the Provider Reimbursement Manual and other documents, but never specifically addressed in regulations. The final rule carries out a directive made in the Omnibus Budget Reconciliation Act of 1989 and addresses changes required by the Omnibus Budget Reconciliation Act of 1990. PMID- 11503757 TI - Facsimile transmission of prescriptions for patients enrolled in hospice programs. Drug Enforcement Administration (DEA), Justice. Final rule. AB - DEA is finalizing, without change, the interim rule with request for comment published in the Federal Register on July 25, 2000 (65 FR 45712). The interim rule amended Title 21, Code of Federal Regulations (CFR) 1306.11(g) to clearly articulate that prescriptions for Schedule II narcotic substances for patients enrolled in hospice care certified by Medicare under Title XVIII or licensed by the state may be transmitted by facsimile. No comments to the interim rule were received. This final rule makes the clarification permanent. PMID- 11503758 TI - Medicaid program; change in application of Federal Financial Participation limits. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule changes the current requirement that limits on Federal Financial Participation (FFP) must be applied before States use less restrictive income methodologies than those used by related cash assistance programs in determining eligibility for Medicaid. This change was originally published as a proposed rule on October 31, 2000 (65 FR 64919). This regulatory change is necessary because the current regulatory interpretation of how the FFP limits apply to income methodologies under section 1902(r)(2) of the Social Security Act (the Act) unnecessarily restricts States' ability to take advantage of the authority to use less restrictive income methodologies under that section of the statute. While the enactment of section 1902(r)(2) of the Act could be read in the limited manner embodied in current regulations the statute does not require such a reading, and subsequent State experience with implementing section 1902(r)(2) of the Act calls into question the current regulation's approach. PMID- 11503759 TI - State child health; implementing regulations for the State Children's Health Insurance Program. Health Care Financing Administration (HCFA), HHS. Final rule. AB - Section 4901 of the Balanced Budget Act of 1997 (BBA) amended the Social Security Act (the Act) by adding a new title XXI, the State Children's Health Insurance Program (SCHIP). Title XXI provides funds to States to enable them to initiate and expand the provision of child health assistance to uninsured, low-income children in an effective and efficient manner. To be eligible for funds under this program, States must submit a State plan, which must be approved by the Secretary. This final rule implements provisions related to SCHIP including State plan requirements and plan administration, coverage and benefits, eligibility and enrollment, enrollee financial responsibility, strategic planning, substitution of coverage, program integrity, certain allowable waivers, and applicant and enrollee protections. This final rule also implements the provisions of sections 4911 and 4912 of the BBA, which amended title XIX of the Act to expand State options for coverage of children under the Medicaid program. In addition, this final rule makes technical corrections to subparts B, and F of part 457. PMID- 11503760 TI - Development and validation of the Hogan Grief Reaction Checklist. AB - The purpose of this article is to provide data on a recently developed instrument to measure the multidimensional nature of the bereavement process. In contrast to widely used grief instruments that have been developed using rational methods of instrument construction, the Hogan Grief Reaction Checklist (HGRC) was developed empirically from data collected from bereaved adults who had experienced the death of a loved one. Factor analysis of the HGRC revealed 6 factors in the normal trajectory of the grieving process: Despair, Panic Behavior, Blame and Anger, Detachment, Disorganization, and Personal Growth. Additional data are provided that support reliability and validity of the HGRC as well as its ability to discriminate variability in the grieving process as a function of cause of death and time lapsed since death. Empirical support is also provided for Personal Growth as an integral component of the bereavement process. The article concludes by considering the substantive as well as psychometric findings of this research for such issues as traumatic grief, anticipatory grief, change in the bereaved person's self-schema, and spiritual and existential growth. PMID- 11503761 TI - Israeli children's reactions to the assassination of the Prime Minister. AB - The assassination of Israel's Prime Minister Yitzhak Rabin by a political opponent caused disbelief, shock, anxiety, and deep grief in the Israeli society. This study reports on 229 4th-grade children's responses to the traumatic event 2 days after it occurred. In the present study, a semi-projective measure, the Bar Ilan Picture Test for Children (R. Itskovitz & H. Strauss, 1982, 1986), was adapted to the situation to assess their affective reactions, social support resources, and coping on the second day after the murder. Overall, the results reveal that their reactions reflect the general adult population affective response to the assassination and resemble the responses of children found in epidemiological studies following trauma and disaster. The role played by the school as social support system is also discussed. PMID- 11503762 TI - Parental bereavement: the crisis of meaning. AB - This descriptive study used qualitative methods to look at two aspects of the search for meaning in parental bereavement--the search for cognitive mastery and the search for renewed purpose. One hundred and seventy-six bereaved parents answered open-ended questions about the experience of their child's death and the meaning of their life since the death. For most parents, the child's death precipitated a severe crisis of meaning and initiated a search for meaning that involved both cognitive mastery and renewed purpose. Those parents who were able to find meaning in the death cited connections with people, the memory of the child, and positive gains resulting from the trauma. The great majority of parents believed that their lives since the death of the child had meaning. Meaning came from connections with people, activities, beliefs and values, personal growth, and connections with the lost child. Implications for grief counseling are discussed. PMID- 11503763 TI - College student bereavement, scholarship, and the university: a call for university engagement. AB - The prevalence of bereavement among traditional-aged college students should impel universities to assist bereaved students on their campuses. Dealing with bereavement can not only challenge a college student's completing the developmental tasks that our society sets for the later adolescent years, but also imperil the student's remaining in school and graduating. It is in the best interests of the university to develop and implement a variety of effective interventions to assist bereaved students. The author argues that universities are communities devoted to scholarly endeavors and should explicitly incorporate the dimension of compassion and caring. An abbreviated case study is used to illustrate the situations in which one grieving student found herself when she returned to school following the death of her father. A call is made for greater university engagement by forming a university-based bereavement center to coordinate and conduct coherent inquiry that fulfills the scholarly functions of discovery, application, and instruction. Four specific actions for a bereavement center are to train nonbereaved students to provide peer support, to provide structured interventions for college students at risk of bereavement complications, to raise consciousness about bereavement on the university campus, and to conduct research into various bereavement populations and bereavement topics. PMID- 11503764 TI - Protection of human research subjects. Department of Health and Human Services (DHHS). Final rule. AB - The Department of Health and Human Services (DHHS) is amending its human subjects protection regulations. These regulations provide additional protections for pregnant women and human fetuses involved in research and pertains to human in vitro fertilization. The rule continues the special protections for pregnant women and human fetuses that have existed since 1975. The rule enhances the opportunity for participation of pregnant women in research by promoting a policy of presumed inclusion, by permitting the pregnant woman to be the sole decision maker with regard to her participation in research, and by exempting from the regulations six categories of research. The rule also provides a mechanism for the Secretary of HHS to conduct or fund research not otherwise approvable after consultation with an expert panel and public review and comment. PMID- 11503765 TI - Opioid drugs in maintenance and detoxification treatment of opiate addiction; Substance Abuse and Mental Health Services Administration, HHS. Final rule. AB - The Department of Health and Human Services and the Substance Abuse and Mental Health Services Administration (SAMHSA) are issuing final regulations for the use of narcotic drugs in maintenance and detoxification treatment of opioid addiction. This final rule repeals the existing narcotic treatment regulations enforced by the Food and Drug Administration (FDA), and creates a new regulatory system based on an accreditation model. In addition, this final rule shifts administrative responsibility and oversight from FDA to SAMHSA. This rulemaking initiative follows a study by the Institute of Medicine (IOM) and reflects recommendations by the IOM and several other entities to improve opioid addiction treatment by allowing for increased medical judgment in treatment. PMID- 11503766 TI - State Vocational Rehabilitation Services Program. Office of Special Education and Rehabilitative Services, Department of Education. Final regulations. AB - The Secretary amends the regulations governing the State Vocational Rehabilitation Services Program. These amendments implement changes to the Rehabilitation Act of 1973 made by the Rehabilitation Act Amendments of 1998 that were contained in Title IV of the Workforce Investment Act of 1998 (WIA), enacted on August 7, 1998, and as further amended in 1998 by technical amendments in the Reading Excellence Act and the Carl D. Perkins Vocational and Applied Technology Education Act Amendments of 1998 (hereinafter collectively referred to as the 1998 Amendments). PMID- 11503767 TI - Percutaneous vertebroplasty. PMID- 11503768 TI - Transcatheter arterial chemoembolization of hepatic tumors. PMID- 11503769 TI - Vacuum-assisted closure for chronic wound healing. PMID- 11503770 TI - Intracoronary brachytherapy for the prevention and management of restenosis. PMID- 11503772 TI - Vaccinations: public health's 'miracle' under scrutiny. PMID- 11503771 TI - Iontophoresis. PMID- 11503773 TI - When faith-healing fails: the Oregon legislature has drawn a fine line between punishing devout parents and ensuring necessary medical care for youngsters. PMID- 11503774 TI - Medicare and Medicaid programs; hospital conditions of participation: anesthesia services. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule amends the Anesthesia Services Condition of Participation (CoP) for hospitals, the Surgical Services Condition of Participation for Critical Access Hospitals (CAH), and the Ambulatory Surgical Center (ASC) Conditions of Coverage Surgical Services. This final rule changes the physician supervision requirement for certified registered nurse anesthetists furnishing anesthesia services in hospitals, CAHs, and ASCs. Under this final rule, State laws will determine which professionals are permitted to administer anesthetics and the level of supervision required, recognizing a State's traditional domain in establishing professional licensure and scope-of-practice laws. States and hospitals are free to establish additional standards for professional practice and oversight as they deem necessary. The hospital anesthesia services CoP, CAH surgical services CoP, and the conforming change to the anesthesia Conditions of Coverage apply to all Medicare and Medicaid participating hospitals, CAHs, and ASCs. PMID- 11503775 TI - Occupational exposure to bloodborne pathogens; needlestick and other sharps injuries; final rule. Occupational Safety and Health Administration (OSHA), Department of Labor. Final rule; request for comment on the Information Collection (Paperwork) Requirements. AB - The Occupational Safety and Health Administration is revising the Bloodborne Pathogens standard in conformance with the requirements of the Needlestick Safety and Prevention Act. This Act directs OSHA to revise the Bloodborne Pathogens standard to include new examples in the definition of engineering controls along with two new definitions; to require that Exposure Control Plans reflect how employers implement new developments in control technology; to require employers to solicit input from employees responsible for direct patient care in the identification, evaluation, and selection of engineering and work practice controls; and to require certain employers to establish and maintain a log of percutaneous injuries from contaminated sharps. PMID- 11503776 TI - Consideration of potassium iodide in emergency plans. Nuclear Regulatory Commission. Final rule. AB - The Nuclear Regulatory Commission (NRC) is amending its emergency planning regulations governing the domestic licensing of production and utilization facilities. The final rule requires that consideration be given to including potassium iodide (KI) as a protective measure for the general public that would supplement sheltering and evacuation. KI would help prevent thyroid cancers in the unlikely event of a major release of radioactivity from a nuclear power plant. The final rule responds to petitions for rulemaking (PRM 50-63 and PRM 50 63A) submitted by Mr. Peter G. Crane concerning the use of KI in emergency plans. PMID- 11503777 TI - Human cells, tissues, and cellular and tissue-based products; establishment registration and listing. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to require human cells, tissue, and cellular and tissue-based product establishments to register with the agency and list their human cells, tissues, and cellular and tissue based products. FDA is also amending the registration and listing regulations that currently apply to human cells, tissues, and cellular and tissue-based products regulated as drugs, devices, and/or biological products. These actions are being taken to establish a unified registration and listing program for human cells, tissues, and cellular and tissue-based products. PMID- 11503778 TI - Occupational injury and illness recording and reporting requirements. Occupational Safety and Health Administration (OSHA), U.S. Department of Labor. Final rule. AB - The Occupational Safety and Health Administration (OSHA) is revising its rule addressing the recording and reporting of occupational injuries and illnesses (29 CFR parts 1904 and 1952), including the forms employers use to record those injuries and illnesses. The revisions to the final rule will produce more useful injury and illness records, collect better information about the incidence of occupational injuries and illnesses on a national basis, promote improved employee awareness and involvement in the recording and reporting of job-related injuries and illnesses, simplify the injury and illness recordkeeping system for employers, and permit increased use of computers and telecommunications technology for OSHA recordkeeping purposes. This rulemaking completes a larger overall effort to revise Part 1904 of Title 29 of the Code of Federal Regulations. Two sections of Part 1904 have already been revised in earlier rulemakings. A rule titled Reporting fatalities and multiple hospitalization incidents to OSHA, became effective May 2, 1994 and has been incorporated into this final rule as Section 1904.39. A second rule entitled Annual OSHA injury and illness survey of ten or more employers became effective on March 13, 1997 and has been incorporated into this final rule as Section 1904.41. The final rule being published today also revises 29 CFR 1952.4, Injury and Illness Recording and Reporting Requirements, which prescribes the recordkeeping and reporting requirements for States that have an occupational safety and health program approved by OSHA under Section 18 of the Occupational Safety and Health Act (the "Act" or "OSH Act"). PMID- 11503779 TI - Determining adjusted income in HUD programs serving persons with disabilities: requiring mandatory deductions for certain expenses; and disallowance for earned income. Office of the Secretary, HUD. Final rule. AB - This final rule amends HUD's regulations in part 5, subpart F, to include additional HUD programs in the list of programs that must make certain deductions in calculating a family's adjusted income. These deductions primarily address expenses related to a person's disability, for example medical expenses or attendant care expenses. The purpose of this amendment is to expand the benefits of these deductions to persons with disabilities served by HUD programs not currently covered by part 5, subpart F. Second, this rule adds a new regulatory section to part 5 to require for some but not all of these same programs the disallowance of increases in income as a result of earnings by persons with disabilities. HUD believes that making these deductions and disallowance available to persons with disabilities through as many HUD programs as possible will assist persons with disabilities in obtaining and retaining employment, which is an important step toward economic self-sufficiency. This rule follows publication of a August 21, 2000 proposed rule, and takes into consideration public comments received on the rule. PMID- 11503780 TI - Medicaid program; Medicaid managed care. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with comment period amends the Medicaid regulations to implement provisions of the Balanced Budget Act of 1997 (BBA) that allow the States greater flexibility by permitting them to amend their State plan to require certain categories of Medicaid beneficiaries to enroll in managed care entities without obtaining waivers if beneficiary choice is provided; establish new beneficiary protections in areas such as quality assurance, grievance rights, and coverage of emergency services; eliminate certain requirements viewed by State agencies as impediments to the growth of managed care programs, such as the enrollment composition requirement, the right to disenroll without cause at any time, and the prohibition against enrollee cost-sharing. In addition, this final rule expands on regulatory beneficiary protections provided to enrollees of prepaid health plans (PHPs) by requiring that PHPs comply with specified BBA requirements that would not otherwise apply to PHPs. PMID- 11503781 TI - Age adjustment using the 2000 projected U.S. population. PMID- 11503782 TI - Abciximab and percutaneous coronary intervention: new indications. Medium-term benefit. AB - (1) Abciximab is now indicated for the prevention of ischaemic complications in patients undergoing percutaneous coronary intervention, and for the prevention of myocardial infarction in patients with unstable angina undergoing percutaneous coronary intervention. (2) The clinical file on abciximab now includes three placebo-controlled trials. (3) The EPIC trial involved patients at high risk of thrombosis, while the EPILOG trial included patients regardless of their risk of thrombosis. In neither trial did abciximab reduce overall mortality. In high risk patients abciximab reduced the risk of a new myocardial infarction at one year, but not for longer. In patients at lower risk, the reduction in the risk of reinfarction persisted for at least six months. (4) The CAPTURE trial showed that abciximab, started 24 hours before angioplasty for unstable angina, reduced the risk of myocardial infarction at one month. (5) The bleeding risk was controlled by the use of heparin at doses adjusted to body weight, and by applying strict precautions to protect the injection site. PMID- 11503783 TI - Paclitaxel and lung cancer: new preparation. No therapeutic progress. AB - (1) Paclitaxel is now licensed, in combination with cisplatin, for the treatment of non small-cell lung cancer in patients not qualifying for surgery or radiotherapy. (2) The clinical file is relatively bulky but of mediocre methodological quality. (3) In one trial the cisplatin + paclitaxel combination was neither more effective nor better tolerated globally than cisplatin monotherapy at a higher dose. (4) The cisplatin + paclitaxel combination has also been compared with the cisplatin + etoposide and cisplatin + teniposide combinations, but showed neither greater clinical efficacy nor fewer adverse effects. PMID- 11503784 TI - Gabapentin monotherapy: new indication. Sometimes helpful. AB - (1) Gabapentin is now licensed for first-line or replacement monotherapy of partial epilepsy, in patients over 12 years of age. (2) The assessment file concurs with current recommendations of medicines agencies. (3) One comparative trial of first-line monotherapy showed a similar efficacy/adverse effects ratio of gabapentin and carbamazepine. Gabapentin has not been compared with valproate sodium. (4) In one trial, involving patients with refractory epilepsy, various doses of gabapentin were added to an ongoing inadequately effective treatment, which was then gradually stopped. Gabapentin monotherapy was considered satisfactory in a minority of patients. (5) The adverse effects of gabapentin are limited to neuropsychological disorders, namely dizzy spells, drowsiness, fatigue and headache. The risk of interactions is also limited. (6) The optimal dose regimen of gabapentin is not yet established. PMID- 11503785 TI - Nabumetone: new preparation. Just another NSAID. AB - (1) Nabumetone is a nonsteroidal antiinflammatory drug recently marketed in France. It has been available in other countries for about 15 years. Its licensed indications cover chronic inflammatory rheumatism and osteoarthritis. (2) The clinical file is bulky, but available trials in chronic inflammatory rheumatism involve only rheumatoid arthritis. (3) In rheumatoid arthritis (4 trials) and osteoarthritis (11 trials), nabumetone was no more effective than other nonsteroidal antiinflammatory drugs with which it was compared. (4) Clinical trial data, pharmacovigilance surveys and epidemiological studies suggest that nabumetone is among the antiinflammatory drugs with the least gastrointestinal adverse effects, but it has not yet been shown that they are less frequent than those of diclofenac, etodolac, ibuprofen or sulindac. PMID- 11503787 TI - Another year... PMID- 11503786 TI - Efavirenz: new preparation. An alternative to HIV protease inhibitors. AB - (1) Efavirenz, a non nucleoside HIV reverse transcriptase inhibitor, is indicated, in combination with other antiretroviral drugs, for the treatment of HIV infection. (2) The methodologically sound clinical file is limited to trials based on validated surrogate end points (viral load) and involving patients in the early stages of HIV infection. (3) Two dose-finding studies favour a dose regimen of 600 mg in a single daily intake for adults. (4) Interim results of three ongoing trials are also available. After 48 weeks of treatment, three-drug regimens comprising efavirenz plus two nucleoside inhibitors were more effective on viral load than three-drug regimens comprising indinavir plus two nucleoside inhibitors. In another trial, after 24 weeks of treatment, a three-drug regimen combining efavirenz and two nucleoside inhibitors was no more effective on viral load than a three-drug regimen comprising nelfinavir and two nucleoside inhibitors. (5) It remains to be determined if four-drug regimens comprising efavirenz, a protease inhibitor and two nucleoside inhibitors are superior to three-drug regimens. (6) Data on the safety profile of efavirenz show mainly neuropsychological and cutaneous disorders. The severity of cutaneous effects in children needs further assessment. PMID- 11503788 TI - Pancytopenia due to the interaction of allopurinol with azathioprine or mercaptopurine. AB - (1) Allopurinol increases the haematological toxicity of azathioprine and mercaptopurine, with a risk of pancytopenia. (2) Combination of allopurinol with azathioprine or mercaptopurine should be avoided. PMID- 11503789 TI - Prohibition of phenolphthalein. AB - Welcome disappearance of phenolphthalein from medicinal use. PMID- 11503790 TI - Management of HIV-infected adults: antiretroviral treatment. AB - (1) The main aim of antiretroviral therapy is to inhibit viral replication as fully and as durably as possible. The reference treatment is currently a regimen combining three antiretroviral drugs belonging to two different classes. (2) Viral load and the CD4+ lymphocyte count are the best available markers for follow-up and decision-making. (3) Viral resistance, along with frequent cross resistance within a given class of antiretroviral drugs, limits the therapeutic options. (4) Adherence to treatment is of major importance. Prescription of antiretroviral drugs must be part of a "treatment plan" decided with and accepted by the patient. (5) First-line treatment and subsequent modifications are based mainly on viral load values. A confirmed rebound of viral replication necessitates a rapid switch in treatment. (6) Viral load rebounds should be managed with antiretroviral drugs that the patient has not yet received and to which no cross-resistance is expected. (7) After repeated treatment failure, T cell counts are more useful and important than viral load values. It may be better to continue the ongoing treatment unchanged than to partially modify it. (8) Treatment success depends partly on the patient's social environment and quality of care. PMID- 11503791 TI - Tamoxifen unsuitable for primary prevention of breast cancer: uncertain efficacy, clear risks. AB - (1) Three large placebo-controlled trials conducted in the United States, the United Kingdom and Italy have tested tamoxifen (20 mg/day) in primary prevention of breast cancer. The British and American trials involved women at particularly high risk. (2) Only the American trial concluded that tamoxifen was effective. The results suggested that treating 130 women with tamoxifen for 5 years would avoid two cases of invasive breast cancer, but at a cost of one severe adverse event (either deep vein thrombosis/pulmonary embolism or uterine cancer). No impact on mortality was documented. (3) No satisfactory reason has yet been forwarded for the conflicting results of the American and British trials. The negative results of the Italian trial are difficult to interpret because of inadequate statistical power. (4) As the preventive effect of tamoxifen is at best limited, and because of uncertainties on the consequences of long-term treatment, prescribing tamoxifen for primary prevention of breast cancer is not justified. (5) Tamoxifen currently has no place in the primary prevention of breast cancer, except in the clinical trial setting. PMID- 11503792 TI - Don't exaggerate the systemic adverse effects of hepatitis B vaccination: the vaccine benefits people at risk. AB - (1) The initial clinical assessment files on hepatitis B vaccines showed no risk of severe systemic adverse effects. (2) Pharmacovigilance data obtained in several countries show that hepatitis B vaccination carries a risk of very rare anaphylactic reactions. (3) Anecdotal observations have raised the worrying possibility that neurological and "autoimmune" conditions may be caused or worsened by hepatitis B vaccination in some patients. Current epidemiological data do not support this possibility, but do not formally exclude it. (4) The risk-benefit ratio of hepatitis B vaccination is advantageous in people at risk. (5) All mass vaccination programmes must include systematic monitoring of adverse effects. PMID- 11503793 TI - Cabergoline and hyperprolactinaemia: new preparation. Better than bromocriptine. AB - (1) Cabergoline, a dopamine agonist already marketed in about 40 countries, is indicated in France for the treatment of hyperprolactinaemia (idiopathic or caused by a pituitary microadenoma). The reference drug in this setting is bromocriptine. (2) The clinical file on cabergoline is methodologically sound, albeit limited mainly to women with amenorrhoea. (3) Two partially blinded comparative trials have shown that cabergoline is significantly more effective than bromocriptine in restarting ovulatory cycles with menstruation. (4) In these trials the incidence of nausea was significantly lower on cabergoline than on bromocriptine. Other adverse events seem to be equally frequent with the two drugs. (5) Cabergoline is effective when taken once or twice a week, while bromocriptine needs to be taken several times a day. PMID- 11503794 TI - Dalteparin new treatment duration: new dosage. Is prolonged prophylaxis needed? AB - (1) In the prevention of deep venous thrombosis after hip surgery, the recommended treatment period with dalteparin has been extended from seven days to 35 days. (2) The clinical file is based on two double-blind trials comparing dalteparin administered for 35 days with dalteparin administered for seven days followed by a placebo, in a total of 546 patients. (3) No clinical trials have compared prolonged dalteparin prevention to dalteparin for one week followed by oral anticoagulant therapy. (4) Clinical vascular events were infrequent between days 7 and 35. There was one death due to pulmonary embolism in the placebo group. One patient on dalteparin died, possibly because of a subdural haematoma discovered at autopsy. (5) Prolonged dalteparin prophylaxis necessitates laboratory monitoring to detect thrombocytopenia. (6) Cost-effectiveness studies of dalteparin administration for 35 days versus dalteparin for seven days followed by oral anticoagulants, are required. PMID- 11503795 TI - Zanamivir: new preparation. No tangible progress against influenza. AB - (1) Zanamivir, an antiviral drug, is marketed in Europe and the United States for treatment of influenza during outbreaks. (2) The clinical file mainly consists of three double-blind, placebo-controlled trials. (3) When administered to patients with symptoms of influenza, at an inhaled dose of 10 mg twice a day, zanamivir has no clear or at least limited benefit, hastening clinical recovery by 0.7-1.5 days, provided that treatment is started within 30 hours of symptom onset. Zanamivir seems to have an action on influenza virus types A and B. No reliable data are available on the impact of zanamivir on overall symptom intensity during the episode or the time required to resume normal activities. The possible value of zanamivir in preventing complications in at-risk subjects remains to be demonstrated. (4) Zanamivir has not been compared with another antiviral drug, amantadine, which has been available for nearly 20 years in France for the same indication. The efficacy of the two drugs regarding the time to recovery is of the same order. Amantadine is effective only on influenza virus type A. (5) In clinical trials the incidence of adverse events was not significantly higher on zanamivir than on placebo. However, cases of bronchospasm following zanamivir administration occurred in patients with asthma and chronic obstructive airways disease. Amantadine, which has been in use for about thirty years, has a relatively good safety profile. (6) Zanamivir (Relenza degrees ) is a costly preparation. PMID- 11503796 TI - Levonorgestrel: new preparation. Emergency contraceptive. AB - (1) The cornerstone of the clinical file on single-agent levonorgestrel (750 mug per tablet) in emergency contraception is a randomised comparative double-blind trial involving nearly 2,000 women. (2) In this trial the efficacy of two levonorgestrel doses (750 mug per dose) was significantly superior to that of two doses of the ethinyloestradiol (100 mug) + levonorgestrel (500 mug) combination: respectively 1.1% and 3.2% of women became pregnant. (3) Nausea and vomiting, dizzy spells and fatigue were half as frequent in the women using single-agent levonorgestrel (750 mug) as in those receiving the combined product. In both groups nearly a third of the women had abnormal periods after treatment. PMID- 11503797 TI - Acellular pertussis vaccines: new preparation. Better tolerated but less active? AB - (1) In France SmithKline Beecham market a three-component acellular pertussis vaccine in the form of a tetravalent vaccine and a pentavalent vaccine. Pasteur M/erieux MSD also market in France a two-component acellular pertussis vaccine in the form of a tetravalent vaccine and a pentavalent vaccine. (2) The two pentavalent vaccines are indicated for an early booster injection at age 16-18 months; the two tetravalent vaccines are indicated for a late booster injection at age 11-13 years. (3) The published assessment file on these vaccines is limited. (4) Immunological studies show that the acellular pertussis valency does not reduce the antigenicity of the valencies with which it is combined (diphtheria, tetanus and poliomyelitis), although there is some doubt regarding the Haemophilus influenzae tybe b valency. (5) In the absence of direct comparisons with the cellular pertussis vaccine available in France in tetravalent or pentavalent vaccines, indirect evidence and data from a trial comparing the acellular two-component vaccine with the cellular vaccine (Vaxicoq degrees ) marketed in France (combined with diphtheria and tetanus valencies) suggest that clinical protection is slightly lower after primary vaccination with the two acellular vaccines. (6) In children, acellular pertussis vaccines are globally better tolerated than the cellular pertussis vaccine. (7) In adolescents, a small study has shown that the three-antigen acellular pertussis vaccine is relatively well tolerated. But the acellular vaccine was released too recently onto the international market to know the precise incidence of rare and severe adverse effects. (8) The new French vaccination schedule for pertussis now includes a booster between 11 and 13 years. Only long-term epidemiological follow-up can show if routine vaccination of adolescents against pertussis will have an impact on the incidence of pertussis in infants and adults. (9) The incidence of Haemophilus b meningitis must continue to be monitored. PMID- 11503798 TI - Trimetazidine: a second look. Just a placebo. AB - (1) Trimetazidine is licensed in France for the prevention of angina attacks and for symptomatic treatment of vertigo and tinnitus. (2) In angina the evidence on trimetazidine is sorely lacking. The only available comparative trial failed to show that trimetazidine monotherapy was even as good as low-dose propranolol. For want of appropriate assessment, it is not known if trimetazidine can reinforce the activity of betablockers, the reference treatment. (3) In symptomatic treatment of vertigo and tinnitus, the three available placebo controlled trials show significant differences in favour of trimetazidine, but these differences are too small to be clinically relevant. (4) Data on the adverse effect profile of trimetazidine are virtually non existent. Gastrointestinal disorders and headache have been reported. PMID- 11503799 TI - Visual field constriction on vigabatrin. AB - (1) Vigabatrin carries a high risk of concentric visual field constriction, sometimes associated with a drop in visual acuity. The changes in the visual field appear to be irreversible. (2) Consequently, vigabatrin can be considered only as a last resort for infantile spasms refractory to steroid therapy and for partial epilepsy refractory to other anticonvulsants. (3) Patients treated with vigabatrin must have their visual fields monitored regularly. PMID- 11503801 TI - Drug-induced stuttering. AB - (1) Remember that some drugs can cause stuttering, even if this side effect appears to be rare. PMID- 11503800 TI - Valve disease and pulmonary arterial hypertension due to appetite suppressants (update). AB - (1) The unfavourable risk-benefit ratio of amphetamine appetite suppressants has led the French medicines agency to withdraw their marketing authorisation. PMID- 11503802 TI - Severe cardiac arrhythmia on fexofenadine? AB - (1) One case of ventricular fibrillation during fexofenadine administration has been reported in a man with a pre-existing long QT interval. PMID- 11503803 TI - Hepatitis B vaccination in countries with low endemicity. AB - (1) Hepatitis B is less frequent in rich countries than in the rest of the world. The prevalence of HBs antigen (a measure of the risk of viral transmission) ranges from less than 0.1% in northern Europe to 1-5% in southern Europe and 8 20% in many African and Asian countries. (2) Initial recommendations to vaccinate only those in high risk groups had no apparent impact on the hepatitis B transmission rate. Changes in behaviour linked to the AIDS pandemic have coincided with a fall in the incidence of acute hepatitis B. (3) The main limitation of immunisation strategies restricted to high risk groups is the fact that no known risk factor is identified in over 30% of cases of acute hepatitis B. (4) All rich countries have recommended routine screening for hepatitis B in pregnant women, and immunisation of newborns whose mothers carry HBs antigen. (5) In countries with low endemicity, HBV transmission usually occurs after the age of 20. Universal vaccination of infants would, therefore, have no effect on the HBV transmission rate for 15-20 years, and then only if the coverage was extensive. Countries in northern Europe, where the prevalence of HBV infection is the lowest in the world, together with Canada, Switzerland and Australia, have not recommended universal immunisation of infants. (6) Universal immunisation of adolescents covers a population closer to the ages most at risk. Only northern European countries have not adopted this policy. PMID- 11503804 TI - Acellular pertussis vaccines: maybe slightly less effective, but better tolerated. AB - (1) With the exception of an American vaccine, whole-cell pertussis vaccines, including the preparation marketed in France, have an efficacy of 90 to 95%. Systemic and local reactions are frequent and can be severe. Cellular vaccines do not seem to cause permanent brain lesions or sudden infant death. (2) Four acellular pertussis vaccines are marketed in France: two have two antigenic components and two have three components. (3) The efficacy of the best acellular pertussis vaccines does not exceed 85% after primary vaccination. Clinical protection may be shorter-lived with acellular than with cellular vaccines. (4) Systemic and local adverse effects, both mild and severe, are significantly less frequent with acellular vaccines. PMID- 11503805 TI - Medicaid program; use of restraint and seclusion in psychiatric residential treatment facilities providing psychiatric services to individuals under age 21. Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This interim final rule with comment period establishes a definition of a "psychiatric residential treatment facility" that is not a hospital and that may furnish covered Medicaid inpatient psychiatric services for individuals under age 21. This rule also sets forth a Condition of Participation (CoP) that psychiatric residential treatment facilities that are not hospitals must meet to provide, or to continue to provide, the Medicaid inpatient psychiatric services benefit to individuals under age 21. Specifically, this rule establishes standards for the use of restraint or seclusion that psychiatric residential treatment facilities must have in place to protect the health and safety of residents. This CoP acknowledges a resident's right to be free from restraint or seclusion except in emergency safety situations. We are requiring psychiatric residential treatment facilities to notify a resident (and, in the case of a minor, his or her parent(s) or legal guardian(s)) of the facility's policy regarding the use of restraint or seclusion during an emergency safety situation that occurs while the resident is in the program. We believe these added requirements will protect residents against the inappropriate use of restraint or seclusion. PMID- 11503806 TI - State Vocational Rehabilitation Services Program. Office of Special Education and Rehabilitative Services, Department of Education. Final regulations. AB - The Secretary amends the regulations governing the State Vocational Rehabilitation Services Program (VR program) by revising the scope of employment outcomes under the VR program. These regulations redefine the term "employment outcome" (as it applies to the VR program) to mean outcomes in which an individual with a disability works in an integrated setting. This action is necessary to reflect the purpose of Title I of the Rehabilitation Act of 1973, as amended (Act), which is to enable individuals with disabilities who participate in the VR program to achieve an employment outcome in an integrated setting. PMID- 11503807 TI - Creating a causal connection: from prenatal drug use to imminent harm. PMID- 11503808 TI - Confidentiality of health information: what can we learn from state partnerships? PMID- 11503809 TI - Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); methodology for coverage of phase II and phase III clinical trials sponsored by the National Institutes of Health. Office of the Secretary; DoD. Final rule. AB - This final rule allows the Department of Defense to waive normal requirements so that covered beneficiaries can participate in Phase II and Phase III clinical trials sponsored or approved by the National Institutes of Health National Cancer Institute (NIH NCI). This waiver authority is expected to promote beneficiary access to promising new treatments and contribute to the development of such treatments. PMID- 11503810 TI - Clopidogrel: new preparation. An alternative to aspirin. AB - (1) Clopidogrel, an antiplatelet drug chemically similar to ticlopidine, is marketed in France for secondary prevention of thrombotic complications in patients with a history of myocardial infarction, ischaemic stroke or peripheral arterial disease. (2) Marketing authorisation was based mainly on the CAPRIE trial, a study that involved 19,815 patients. In this trial of secondary cardiovascular prevention, clopidogrel was slightly more effective than aspirin (325 mg/day) according to a statistical analysis of a combined end point (ischaemic stroke, or myocardial infarction, or death of vascular causes). The difference was more marked in the subgroup of patients with obstructive arterial disease of the lower limbs. (3) Clopidogrel was well tolerated in this trial. The only adverse effects more frequent on clopidogrel than on aspirin were rash and diarrhoea. (4) Clopidogrel showed no haematological toxicity, an adverse effect that restricts the use of ticlopidine. (5) The lack of long-term follow up in real clinical settings prevents any meaningful estimation of the safety profile or of the risk of drug interactions. PMID- 11503811 TI - Raloxifene: new preparation. Not better than oestrogen. AB - (1) Raloxifene is marketed in France for the prevention of non traumatic vertebral fracture in postmenopausal women. In animal pharmacology studies it was found to both agonise and antagonise oestrogen. (2) The assessment file is methodologically sound but fails to answer a good number of practical questions. (3) A placebo-controlled trial showed that raloxifene reduced the risk of vertebral collapse after two years of treatment, in both the primary and secondary prevention settings, but no effect was demonstrated on non vertebral fractures. Furthermore, raloxifene reduced the risk of breast cancer in this trial. (4) Two trials versus combined hormone replacement therapy showed a more favourable effect of the latter on surrogate end points reflecting the risk of fracture and the cardiovascular risk (changes in bone mineral density and lipid profile). (5) Compared with combined hormone replacement therapy, raloxifene reduced the incidence of menorrhagia and mastodynia, but did not relieve symptoms linked to the menopause. (6) The results of animal studies call for close clinical monitoring to detect a possible increase in the incidence of ovarian cancer. PMID- 11503812 TI - Entacapone: new preparation. Comparative data are lacking. AB - (1) Entacapone, a catechol-O-methyltransferase inhibitor, is indicated, in combination with levodopa + a dopadecarboxylase inhibitor, for the treatment of parkinsonian patients who have motor fluctuations on levodopa therapy. (2) The clinical file contains only placebo-controlled trials. (3) The two main clinical trials involved patients with moderate fluctuations while on levodopa, usually combined with other antiparkinsonian drugs. They showed a moderate effect of entacapone, with an increased duration of motor improvement ("on" periods) of approximately one hour, but one trial showed no increase in overall patient satisfaction. (4) In clinical trials the main adverse effects of entacapone were dyskinesias and gastrointestinal disorders (especially diarrhoea and abdominal pain). Given the limited number of patients included in these clinical trials, the risk of rare but severe adverse effects (especially hepatitis) cannot be ruled out. (5) Entacapone therapy can cost up to four times more than treatment with bromocriptine. PMID- 11503813 TI - Nifuroxazide in acute diarrhoea: OTC preparation. Irrational. AB - (1) Nifuroxazide, an intestinal antibacterial agent, is now available in France, without a prescription, for the treatment of acute diarrhoea in adults. (2) According to the only available comparative randomised trial, there is no effect on dehydration. Relative to a placebo, the mean number of stools is reduced by about one per day during the first two days of treatment, with no significant difference thereafter. PMID- 11503814 TI - Perindopril + indapamide: new preparation. Simple trick. AB - (1) The combination of perindopril 4 mg + indapamide 1.25 mg is approved for second-line treatment of hypertension after failure of perindopril alone. (2) The other combination, of a low dose of an angiotensin-converting-enzyme inhibitor (2 mg of perindopril) and a diuretic (0.625 mg of indapamide), is being promoted as first-line treatment of hypertension. (3) The clinical files for both preparations are limited to the strict minimum. (4) A dose-finding study showed that the perindopril 4 mg + indapamide 1.25 mg dose combination offered the best risk-benefit ratio by comparison with combinations containing the same perindopril dose but other indapamide doses. (5) A double-blind trial suggests that the antihypertensive activity of the perindopril 4 mg + indapamide 1.25 mg combination is equivalent to that of the captopril 50 mg + hydrochlorothiazide 25 mg and enalapril 20 mg + hydrochlorothiazide 12.5 mg combinations. The safety profile was the same for the three combinations. (6) The 2-mg perindopril combination has not been compared with perindopril monotherapy at the usual dose of 4 mg, or with indapamide monotherapy at a mean dose of 2.5 mg. (7) The two combinations are costlier than their competitors. PMID- 11503816 TI - Penciclovir: new preparation. Slightly effective. AB - (1) A penciclovir skin cream has been approved in France for local treatment of herpes simplex labialis. (2) The clinical file is limited to a single trial versus the excipient. This double-blind trial showed that, even when started immediately after the first manifestations occur, penciclovir cream failed to prevent the emergence of skin lesions. The only effect was a reduction in the duration of the lesions and associated pain by a few hours. In the absence of any specific assessment, there is no evidence that penciclovir improves overall patient satisfaction. (3) Penciclovir has not been compared with aciclovir skin cream. PMID- 11503815 TI - Levonorgestrel intrauterine device: new preparation. An alternative. AB - (1) Mirena degrees is a contraceptive intrauterine device delivering levonorgestrel. (2) Its assessment is based on two prospective randomised multicenter trials comparing it to a copper device. (3) It is not known whether it is any more difficult to insert than other devices. (4) Efficacy, in terms of absolute protection and risk of extrauterine pregnancy, appears to be very good, at least equal to that of copper devices. (5) Women were less subject to menorrhagia than were women using a copper device. (6) In clinical trials up to a third of women had amenorrhoea. Some women had spotting during the first three months, and other adverse effects (e.g. acne, headache and mastodynia) were more frequent than among women using copper devices. PMID- 11503817 TI - Anaemia due to ACE inhibitors and losartan in patients with renal failure. AB - (1) Angiotensin-converting-enzyme inhibitors (ACE inhibitors) can cause or aggravate anaemia in patients with chronic renal failure (especially those on dialysis) and in renal graft recipients. (2) In these settings, ACE inhibitor therapy can increase epoetin consumption. (3) Similar reports have been made with losartan, an angiotensin II receptor antagonist. (4) The risk-benefit ratio of ACE inhibitors and angiotensin II receptor antagonists in patients with chronic renal failure should be regularly reassessed. PMID- 11503818 TI - Poppers intoxication. AB - (1) Should be considered in cyanosis caused by methaemoglobinaemia. PMID- 11503819 TI - Neurological and cardiac events on phenylpropanolamine. AB - (1) Phenylpropanolamine, or norephedrine, is a sympathomimetic vasoconstrictor very similar to amphetamine, and carries a risk of potentially severe cardiac, neurological and psychiatric events. (2) One should be aware of the presence of phenylpropanolamine in many non prescription drugs. (3) Pharmacists should pay special attention to the presence of phenylpropanolamine in freshly prepared slimming mixtures. PMID- 11503820 TI - Neurological disorders after zidovudine exposure in utero. AB - (1) New cases of mitochondrial disorders have been described in French children exposed to zidovudine in utero. Despite this, zidovudine remains the reference prophylaxis for mother-child HIV transmission. PMID- 11503821 TI - Headache due to antimigraine 5HT1 receptor agonists. AB - (1) Like other analgesics and antimigraine drugs, the 5HT1 receptor agonists, or triptans, can cause a self-sustaining headache syndrome. (2) Headache due to 5HT1 receptor agonists should be suspected when a patient presents with an increased frequency of migraine or non migraine headache associated with daily intake of such drugs. Withdrawal usually leads to the disappearance of those headaches that are due to the drug, or at least a reduction in their frequency. PMID- 11503822 TI - Choosing a macrolide: drug interactions must be taken into account. AB - (1) Some macrolides carry a strong risk of potentially severe drug interactions. (2) Erythromycin is the macrolide with the biggest risk of interactions. (3) Spiramycin is the macrolide with the smallest risk of interactions. Azithromycin and dirithromycin also seem to have a low risk of interactions, but they are far more recent and therefore less well known. (4) In first-line use, unless there are specific indications, a macrolide with a low risk of interactions should be used. PMID- 11503823 TI - Consumption and dangers of ecstasy: an amphetamine derivative with a misleading reputation of safety. AB - (1) Ecstasy carries a risk of potentially severe short-term complications, even after occasional ingestion of low doses. The risk has not been assessed thoroughly, however, and long-term toxicity is even less well established. PMID- 11503824 TI - Our correspondence with the World Health Organization about the hypertension guidelines. AB - (1) Following publication of the hypertension guidelines by the World Health Organization (WHO) and the International Society of Hypertension (ISH) we wrote to Dr Brundtland, criticizing the recommendations that markedly conflicted with reliable published data. (2) We were not happy with Dr Brundtland's response to our letter, so we have stopped the process to become a WHO collaborating centre. PMID- 11503825 TI - Pamidronic acid and Paget's disease: new indication. Short-course treatment. AB - (1) Pamidronic acid, a diphosphonate, is now approved for intravenous treatment of Paget's disease in France. Treatment is given over two to three days, in hospital, and can be repeated six months later. (2) The clinical file contains only non comparative trial data suggesting that a total dose of 180-210 mg has a clinical benefit in most patients and tends to normalise biochemical criteria thought to reflect disease progression. (3) Because no comparative trials have been done, there is no way of knowing whether a short course of intravenous pamidronic acid is any more or less effective that long-term oral etidronic acid or tiludronic acid therapy. However, in the non comparative trials a number of patients who had become resistant to oral diphosphonate therapy had a clinical and/or biochemical response to intravenous pamidronic acid. (4) While intravenous pamidronic acid is well tolerated overall, the precise incidence of its adverse effects on bone (transient increase in pain, osteomalacia) and the eyes (uveitis) is not known. PMID- 11503827 TI - Rituximab: new preparation. Encouraging preliminary results. AB - (1) Rituximab is indicated for the treatment of patients with relapsed or chemoresistant stage III-IV follicular lymphoma. (2) The clinical file is currently limited to 300 patients treated in non comparative trials. (3) Four infusions of rituximab, at a dose of 375 mg/m2/week, led to at least a partial tumour response in 50% of these poor-prognosis patients, and prevented disease progression for more than nine months in the responders (median value not yet reached). (4) Adverse reactions are frequent and sometimes severe during the first infusion. PMID- 11503826 TI - Codeine for child pain: new preparation. Helpful in some cases. AB - (1) This codeine-based syrup is indicated for the treatment of pain in children. It is the first step 2 analgesic (WHO classification) to become available in France for oral treatment of children. (2) Efficacy and the optimal dose regimen are based mainly on lengthy experience with codeine in other countries. Only a few small trials on acute pain are available. (3) The safety profile is that of all opiates: adverse effects comprise mainly constipation, nausea and vomiting. (4) Accidental ingestion by a young child can lead to severe poisoning. PMID- 11503828 TI - Etidronic acid and steroid therapy: new indication. No proven impact on fractures. AB - (1) Etidronic acid (disodium etidronate) has a new licensed indication in the prevention of bone loss due to steroid therapy. (2) Three randomised, double blind placebo-controlled trials have been done in this setting. The benefit of etidronic acid has been shown only on the basis of lumbar bone density (a surrogate end point) and only in women. (3) There is no concrete evidence that etidronic acid is any more effective than a placebo in reducing the risk of fracture in patients on steroid therapy. Similarly, in the absence of comparative trials there is no proof that etidronic acid is any more effective than hormone replacement therapy in postmenopausal women. PMID- 11503829 TI - Octreotide and bleeding oesophageal varices: new indication. Nothing new. AB - (1) Octreotide, a synthetic somatostatin analogue, is now also indicated for emergency treatment of ruptured oesophageal varices pending endoscopic treatment. (2) The assessment file in this indication is limited. (3) The two available comparative trials show no difference in efficacy compared with terlipressin, but their statistical power was low. Octreotide has not been compared with somatostatin, the efficacy of which is not clearly established either. (4) A placebo-controlled trial, published only as an abstract, showed no specific activity of octreotide on bleeding oesophageal varices. (5) There are few reports of adverse effects with octreotide. There is nothing to suggest that its adverse effects differ from those of somatostatin, when the drug is used in patients who cannot tolerate terlipressin. Octreotide is no easier to administer than somatostatin or terlipressin. PMID- 11503830 TI - Valve disease due to appetite suppressants (cont'd). AB - (1) The risk of cardiac valve disease linked to appetite suppressants has now been confirmed by several studies. It seems to increase with the duration of treatment. (2) The outcome after treatment cessation is poorly known. (3) Patients exposed to these drugs should continue to be monitored. PMID- 11503831 TI - Cholesterol-lowering drugs. Some drugs with demonstrated efficacy but different benefits in primary and secondary prevention. AB - (1) In primary prevention trials, pravastatin and lovastatin prevented myocardial infarction and had a positive risk-benefit ratio in men with LDL-cholesterol values exceeding 4.5 mmol/l (1.7 g/l). Cholestyramine and gemfibrozil also prevented myocardial infarction in men with more severe hypercholesterolaemia; while clofibrate had a negative risk-benefit ratio in patients with moderate hypercholesterolaemia. These treatments have not been assessed for primary prevention in women or in patients aged over 70. (2) In trials involving patients with coronary heart disease, pravastatin and simvastatin both reduced the risk of myocardial infarction and/or mortality in patients of both sexes with LDL-cholesterol values above 3.2 mmol/l (1.2 g/l). Gemfibrozil also reduced the risk of myocardial infarction but not mortality, while clofibrate and bezafibrate had no preventive effect. PMID- 11503832 TI - Flawed WHO recommendations on hypertension: WHO has damaged its reputation. AB - (1) The international expert group ignored the ground rules governing clinical assessment, and clinical trial data too, taking up a stance that favours drug manufacturers. PMID- 11503833 TI - Buprenorphine replacement therapy: a confirmed benefit. AB - (1) Three years after high-dose buprenorphine preparations were first marketed in France, we examine their use as replacement therapy for heroin addiction. (2) Various surveys of community pharmacists have shown that the prescribing and dispensing conditions are feasible in the routine ambulatory setting. However, teamwork between physicians and pharmacists is rarely optimal, and fractionated dispensing is under-used. This could lead to abuse by a minority of patients. (3) During ambulatory management, at a mean dose of 8 mg/day sublingually, long term buprenorphine therapy seems to yield a reduction in drug consumption by most patients, and can help with social reintegration. (4) The risks of buprenorphine treatment are mainly linked to abuse (massive doses, intravenous injection). Fatalities have occurred after high doses of buprenorphine combined with benzodiazepines, especially when taken intravenously. (5) The limited data on buprenorphine intake during pregnancy are reassuring. PMID- 11503834 TI - Orlistat: new preparation. No hurry . . . AB - (1) Treatments for obesity are disappointing. None has yet shown an effect on morbidity and mortality. Non drug treatments are poorly assessed. Stable long term weight loss necessitates long-term management. (2) Orlistat, a gastrointestinal lipase inhibitor, is indicated, in combination with a low calorie diet, for the management of obesity. (3) The assessment file is rather bulky and methodologically sound, at least in terms of the "weight loss" end point. (4) During medium-term trials (12-24 months), orlistat administered at a dose of 120 mg three times a day and combined with dietary intervention had a moderate positive impact on body weight (-3.5 kg on average). (5) No longer-term trials have been done. (6) It is not known whether this drug affects morbidity and mortality linked to obesity. (7) In clinical trials there was an increase in the frequency of breast cancer among patients treated with orlistat. This potential risk is currently being assessed in a specific trial. (8) Gastrointestinal adverse effects are frequent. (9) Treatment is costly. PMID- 11503835 TI - Montelukast: new preparation. No current use in asthma. AB - (1) Montelukast, an antiasthmatic drug belonging to the leukotriene antagonist family, has two indications in France: as adjunctive treatment for mild to moderate chronic asthma when regular inhaled steroid therapy and short-acting inhaled beta 2 stimulants "on demand" are inadequate; and in the prevention of effort-induced asthma. (2) The clinical file on montelukast contains no methodologically acceptable comparisons with reference treatments. (3) Several placebo-controlled trials have shown the efficacy of montelukast, with an improvement in clinical scores and respiratory function tests in chronic asthma; and prevention of effort-induced asthma. (4) In chronic asthma montelukast has not been compared with oral or inhaled long-acting beta 2 stimulants, or with sustained-release theophylline in patients inadequately controlled by steroid therapy. (5) In effort-induced asthma, only two trials have compared montelukast to salmeterol. On the basis of preliminary results the authors concluded that montelukast was superior in both studies. (6) Clinical trials showed no clear difference in the frequency of side effects in patients on montelukast and those on a placebo. However, montelukast may possibly be associated with the Churg and Strauss syndrome in rare cases. (7) Montelukast is an expensive drug. PMID- 11503836 TI - Olanzapine: new preparation. Keep an eye on this neuroleptic. AB - (1) Olanzapine, a neuroleptic, has obtained European marketing authorisation for the treatment of schizophrenia. (2) The clinical file is satisfactory, but in the absence of relevant trials it has not yet been demonstrated that olanzapine has a specific activity on the positive or negative symptoms of schizophrenia. (3) The global efficacy of olanzapine was not significantly different from that of haloperidol in two of the three comparative trials published to date. (4) The only relevant comparative trial fails to demonstrate the superiority of olanzapine over risperidone. (5) Olanzapine has fewer adverse neurological effects than haloperidol, but there is no evidence that it differs from other recent neuroleptics in this respect. (6) Olanzapine can have anticholinergic adverse effects and frequently causes weight gain. (7) Active pharmacovigilance is required, as subclinical cases of elevated transaminase levels, increased blood pressure and QT prolongation were observed in clinical trials (2,500 patients treated). PMID- 11503837 TI - Tretinoin and cutaneous photoageing: new preparation. Guaranteed adverse effects! AB - (1) A cream containing 0.05% of tretinoin is approved in France for the treatment of sun-induced skin damage ("photoageing"). (2) Three comparative trials versus the excipient show that the effects of the tretinoin cream are at best limited and slow to occur; furthermore, they disappear on treatment cessation, necessitating long-term use. (3) The 0.05% tretinoin cream has poor local tolerability: most subjects develop irritation and fragile skin, necessitating a longer interval between each application. Systemic adverse effects can occur in some circumstances. (4) There are persistent doubts over safety during pregnancy. PMID- 11503838 TI - Topiramate: new indication. A bulkier assessment file. AB - (1) The licensed indications of topiramate have been extended to cover treatment (in combination with an inadequately effective ongoing treatment) of generalised tonic-clonic epilepsy in adults and children over four years, and partial epilepsy in children over four. (2) The efficacy of topiramate was established in two double-blind, placebo-controlled trials in patients with generalised tonic clonic epilepsy, and in a double-blind, placebo-controlled trial in patients with partial epilepsy. (3) Topiramate has not been compared with other antiepileptics with similar indications, i.e. lamotrigine and vigabatrin. (4) Topiramate appears to be relatively well tolerated. PMID- 11503839 TI - Ticlopidine: a second look. No further use in routine practice. AB - (1) The clinical file on ticlopidine is based mainly on a placebo-controlled trial involving patients with lower-limb arterial disease, and two large double blind trials in the post-stroke period, in which the comparator was a placebo in one and aspirin in the other. (2) Ticlopidine proved more effective than the placebo in both indications. In secondary prevention of ischaemic stroke, ticlopidine has a moderate advantage over aspirin in terms of efficacy but carries a risk of serious adverse effects, especially haematological. (3) An indirect comparison suggests that clopidogrel, another antiplatelet drug chemically close to ticlopidine, has comparable efficacy in cardiovascular prevention and no severe adverse effects to date. PMID- 11503840 TI - Late cardiotoxicity of anthracyclines. AB - (1) Cardiotoxicity of anthracyclines increases with the cumulative dose and can lead to heart failure. (2) Heart failure can occur several years after treatment, even in patients previously free of cardiac manifestations. (3) Late cardiac effects can be observed in patients who have received less than the recommended maximal cumulative dose. (4) There is no validated monitoring procedure. PMID- 11503841 TI - Management of type 2 diabetes: long-awaited evidence of benefits after blood sugar control. AB - (1) The UKPDS trial was a very large, complex, comparative study with methodological weaknesses such as the absence of blinding. It showed that lowering the blood glucose level in patients with type 2 diabetes reduces the risk of clinical complications, especially diabetic microangiopathy. (2) In contrast, glycaemic control had no statistically significant impact on mortality. (3) Contrary to findings in a previous trial, glucose-lowering sulphonylureas and insulin did not increase cardiovascular morbidity or mortality in the UKPDS study. (4) Glibenclamide was the only drug tested that yielded a statistically significant reduction in the risk of clinical complications linked to type 2 diabetes. (5) Strict glycaemic control with a glucose-lowering sulphonylurea or insulin was associated with hypoglycaemic episodes in approximately a quarter of patients each year. (6) Metformin gave conflicting results that are difficult to explain: metformin reduced mortality in overweight patients with type 2 diabetes; but in diabetic patients poorly controlled by glucose-lowering sulphonylureas, mortality was higher in the group treated with the sulphonylurea + metformin combination than in the group that continued treatment with a sulphonylurea alone. (7) Currently, the results of the UKPDS trial are the only available clinical data on which to base the choice of treatment for type 2 diabetic patients aged between 25 and 65 years. When a glucose-lowering drug is considered necessary and is not contraindicated, the first-line choice is glibenclamide for diabetics who are not overweight, and metformin for those who are. PMID- 11503842 TI - Fractures in adults on systemic steroid therapy: which prophylaxis? AB - (1) Systemic steroid therapy leads to a loss of bone density after a few months. The loss is at least partly reversible on treatment cessation. Together with age, the underlying disease, and reduced mobility, systemic steroid therapy is a risk factor for fractures. (2) There are no treatments with proven efficacy in the prevention of fractures among patients on systemic steroid therapy. Prevention is thus based on restricting steroid therapy to situations where the benefits are likely to outweigh the risks. (3) The first preventive measure is to encourage adequate calcium intake, as for all subjects at risk of osteoporosis. There is no firm evidence that all patients on steroids require medicinal calcium supplementation. (4) Some treatments slow the decline in bone density associated with steroid therapy, but none has a demonstrated preventive effect on symptomatic fractures. This is the case of the calcium + vitamin D combination, which has the best risk-benefit ratio. Two diphosphonates and, in postmenopausal women, hormone replacement therapy, also have a positive effect on bone density. PMID- 11503843 TI - Serotonin reuptake inhibitor antidepressants and pregnancy: many unanswered questions. AB - (1) Serotonin reuptake inhibitor antidepressants are not teratogenic in animals. (2) Human data are limited, and most involve fluoxetine. (3) No data pointing to a teratogenic effect after fluoxetine exposure during the first trimester have been reported. (4) Self-resolving neurological signs have been observed in newborns exposed to fluoxetine at the end of pregnancy. (5) The paucity of data requires all prescribers of antidepressants to pregnant women to report any adverse events occurring in a child or mother to a teratogenicity or pharmacovigilance centre. PMID- 11503844 TI - Self-care among older adults: an analysis of the objective and subjective illness contexts. AB - OBJECTIVES: The authors hypothesize that older adults diagnosed with arthritis show a greater reliance on objective factors in their self-care behaviors, whereas those diagnosed with heart problems or hypertension demonstrate a greater reliance on more general belief-laden factors. METHODS: A total of 794 older adults (mean age = 69.3) who were professionally diagnosed with arthritis, heart problems, or hypertension completed a telephone survey about a number of aspects of their illness condition and their general well-being. RESULTS: The results from the hierarchical regression analyses indicate that objective factors and illness-specific beliefs are better predictors of self-care behavior in the arthritis group, whereas general beliefs (e.g., self-efficacy and general well being) are better predictors of such behavior in the heart problems and hypertension groups. DISCUSSION: The analyses support the authors' hypothesis. The results are discussed in the context of expanding the Health Belief Model of self-care. PMID- 11503845 TI - Valuation of life: a concept and a scale. AB - OBJECTIVES: The objective was to derive and test the psychometric characteristics of a scale to measure Valuation of Life (VOL). METHODS: Four samples were used in successive phases of exploratory factor analysis, confirmatory factor analysis, reliability and validity testing, and exploration of response-error effects. Estimates of Years of Desired Life were obtained under a variety of hypothetical quality-of-life (QOL)-compromising conditions of poor health. RESULTS: Confirmed 13-item (Positive VOL) and 6-item (Negative VOL) factors were obtained. A significant relationship between VOL and most Years of Desired Life estimates remained when demographic, health, quality of life, and mental health measures were controlled. Analysis of Negative VOL revealed that some respondents misunderstand the meaning of an agree response to negatively phrased items. DISCUSSION: VOL is a cognitive-affective schema whose function as a mediator and moderator between health and end-of-life decisions deserves further research. PMID- 11503846 TI - The Danish 1905 cohort: a genetic-epidemiological nationwide survey. AB - OBJECTIVES: The authors studied nonagenarians, a rapidly growing age group whose cognitive and physical abilities have yet to be investigated systematically. METHODS: All Danes born in 1905 were invited to participate in a home-based 2 hour multidimensional interview, including cognitive and physical performance tests and collection of DNA, carried out by lay interviewers. Population-based registers were used to evaluate representativeness. RESULTS: There were 2,262 participants. A total of 1,632 (72%) gave a DNA sample. Participants and nonparticipants were highly comparable with regard to marital status, institutionalization, and hospitalization patterns, but men and rural area residents were more likely to participate. Six months after the survey began, 7.2% of the participants and 11.8% of the nonparticipants had died. DISCUSSION: Despite the known difficulties of conducting surveys among the extremely old, it was possible to conduct a nationwide survey, including collection of DNA, among more than 2,000 fairly nonselected nonagenarians using lay interviewers. PMID- 11503847 TI - The impact of family environment and decision-making satisfaction on caregiver depression: a path analytic model. AB - OBJECTIVES: This research examines caregiver depression in the context of traditional care-related primary stressors, such as the caregiving context and care-recipient impairment, and secondary stressors, such as family environment and decision-making satisfaction. METHODS: The authors examine a causal (path) model of depression among 244 caregivers. Ordinary least squares regression results are used to determine the direct and indirect effects of stressors on caregiver depression. RESULTS: The path coefficients obtained show that adaptability and conflict have the most powerful net effects. With the caregiving context variables, they explain approximately 30% of the variance in decision-making satisfaction. Family adaptability and decision-making satisfaction also have significant paths. The caregiving context, network, family environment, and decision-making variables explain approximately 25% of the variance in caregiver depression. DISCUSSION: These findings suggest that practitioners working with caregivers to ameliorate depression need to examine the broader aspects of family environment and caregiver perceptions related to decision making. PMID- 11503848 TI - A molecular view of coping behavior in older adults. AB - OBJECTIVES: The purpose of this study was to assess age, gender, ethnicity, and education differences in specific ("molecular") coping behaviors for three older adult age groups. METHODS: Thirty-five specific coping reactions were assessed on the item level for 74 sexagenarians, 70 octogenarians, and 116 centenarians of the Georgia Centenarian Study. RESULTS: A multivariate analysis of covariance revealed significant age group, gender, and education differences for 14 coping reactions. Four items were affected exclusively by age; five were affected only by gender; and three were affected only by education. One item showed age group and education differences; another showed gender and education differences. No ethnicity differences were obtained. The largest effect for an age group difference was found for accepting health problems. DISCUSSION: The findings suggest that a molecular view of specific coping behaviors in reaction to health problems, in addition to global measures of coping, is essential. PMID- 11503849 TI - Contribution of education to health and life satisfaction in older adults mediated by negative affect. AB - OBJECTIVES: The authors developed a model of relationships between two enduring attributes (educational attainment and negative affect) and two indicators of successful aging (health and life satisfaction). METHODS: A probability sample of 1,177 participants (age 55 and older) were interviewed four times at 6-month intervals. Structural equation models were developed based on the authors' hypothetical model proposing a mediating effect of negative affect between health and successful aging. RESULTS: As predicted, education and negative affect both were directly related to health and life satisfaction. Also, as predicted, negative affect mediated the relationship between education and successful aging indicators. DISCUSSION: Education appears to confer a lifelong advantage for healthy aging. Part of this advantage is accounted for by the relationship between education and trait negative affect. Higher educational attainment is related to lower levels of trait negative affect; lower negative affect results in better health and life satisfaction. PMID- 11503850 TI - Botulinum toxin type A and dynamic equinus in children with cerebral palsy: new indication. Better than repeat casts. AB - (1) Botox degrees , a product based on type A botulinum toxin, has received a new licensed indication in the local treatment of dynamic equinus in children with spasticity due to cerebral palsy. (2) Three placebo-controlled trials show that intramuscular injections of type A botulinum toxin reduce spastic equinus and substantially improve walking for at least 3 months. Two small trials, each involving 20 children, show no difference in effects between type A botulinum toxin and successive stretching casts. (3) In this setting the risk of adverse effects is smaller with type A botulinum toxin than with stretching casts. (4) Treatment with type A botulinum toxin is costly. PMID- 11503851 TI - Hypnotic dependence: zolpidem and zopiclone too. AB - (1) Loss of efficacy can occur after a few weeks of treatment with zolpidem and zopiclone. This is not a reason to increase the dose. (2) Zolpidem and zopiclone carry a risk of dependence and of potentially severe disorders (e.g. seizures) during withdrawal. (3) The risk-benefit ratio of benzodiazepines and related drugs (zopiclone and zolpidem) remains more favourable than that of other drugs used in insomnia. However, first-line treatment of sleep disorders should not necessarily be drug-based. PMID- 11503852 TI - Erythromycin-induced pyloric stenosis in infants. AB - (1) Several cases of pyloric stenosis have occurred in infants treated with erythromycin during the first weeks of life. (2) A disturbing report of a cluster of cases is enough to avoid erythromycin use in neonates. PMID- 11503853 TI - Treatment of chronic hepatitis B: interferon alfa first. AB - (1) Chronic hepatitis B is defined by the persistence of circulating HBs antigen for more than 6 months. Between 15 and 25% of chronic HBV carriers die prematurely of complications (mainly cirrhosis and hepatocellular carcinoma). (2) Patients with chronic hepatitis B rarely clear the virus spontaneously, but viral replication ceases in approximately 10% of patients annually, with disappearance of HBe and viral DNA, and emergence of anti-HBe antibodies. (3) Active viral replication and histologically proven liver necrosis are risk factors for progression to cirrhosis. (4) Antiviral treatments have been assessed only in patients with active viral replication. (5) Interferon alfa has been widely tested in the clinical setting. In one trial, in which patients were followed for 7 years, mortality was lower in the treatment group than in untreated controls. (6) Interferon alfa must be injected, and its administration may be followed by adverse effects such as a 'flu-like syndrome (frequently), psychiatric problems and potentially severe thyroid disorders. (7) Interferon alfa monotherapy for 4-6 months (possibly extended to 8-9 months) remains the first-line treatment for chronic hepatitis B. (8) Lamivudine has documented antiviral efficacy but its effect is only temporary in many patients. In the only trial comparing lamivudine with interferon alfa, lamivudine was no more effective than interferon in the short term (on the basis of serological and histological end points), despite a bias in its favour. Trials of lamivudine + interferon alfa in patients who fail to respond to interferon monotherapy have given unfavourable results. (9) Adverse effects are infrequent on lamivudine, but pharmacovigilance is required to assess potential hepatic and pancreatic effects at the dose used in this indication. (10) The long-term effects of lamivudine are unknown, especially on the risks of cirrhosis, hepatocarcinoma and the selection of resistant mutants. (11) Pending further data, lamivudine should be used only in clinical trials and cohort studies. PMID- 11503854 TI - Antihypertensives after 70. The STOP-2 trial confirms key place for diuretics. AB - (1) Diuretics remain the first-line antihypertensive for elderly patients. PMID- 11503855 TI - Behaviour therapy for chronic insomnia. AB - (1) In chronic insomnia, cognitive-behaviour treatments appear to be as effective as benzodiazepines. PMID- 11503856 TI - No efficacy of homeopathy against influenza. AB - (1) A meta-analysis stressing the poor methodological quality of trials of homeopathic preparations in influenza concludes that there is no proven preventive or curative effect. PMID- 11503857 TI - Drug interactions with selective serotonin reuptake inhibitors, especially with other psychotropics. AB - (1) Selective serotonin reuptake inhibitors (SSRIs) are involved in many drug interactions with potentially serious clinical consequences. (2) These interactions involve all SSRIs but particularly fluoxetine, which is the best studied antidepressant in this family. (3) Because of their long elimination half-life (particularly fluoxetine) the risk of interactions persists for several days or even weeks after SSRI withdrawal. (4) Drug interactions with clinical consequences usually involve combinations of an SSRI with other psychotropics, especially monoamine oxidase inhibitor (MAOI) and tricyclic antidepressants, clozapine, lithium, methadone, etc. (5) The clinical consequences of drug interactions with SSRI are either due to overdosing of the drug combined, or to a serotonin syndrome with neuromuscular and vegetative (autonomic) symptoms. (6) Interactions with a number of other drugs have been reported, especially carbamazepine, phenytoin and oral anticoagulants, with a risk of overdose of these drugs. (7) The risk of hyponatraemia linked to SSRIs seems to be increased during concomitant treatment with diuretics. PMID- 11503858 TI - Medroxyprogesterone and palliative care: new indication. No impact on quality of life. AB - (1) Oral medroxyprogesterone has been granted a new licence in France for the treatment of weight loss and loss of appetite in patients receiving palliative care for cancer. (2) The clinical file on medroxyprogesterone in this indication is limited. A large number of patients were excluded from the final assessments in the three published placebo-controlled trials. (3) Overall, these trials show that 12 weeks of treatment with medroxyprogesterone 300-1,000 mg a day orally is associated at best with median weight gain of 3 kg, or weight stabilisation, but with no effect on quality of life. (4) Although no cases of thrombosis were reported in the clinical trials, the risk of thrombosis cannot be ruled out in medroxyprogesterone therapy. PMID- 11503859 TI - Anti-D human immunoglobulin: new preparation. Important in young Rh D (-) women. AB - (1) Nat/eaD degrees , a product based on anti-D human immunoglobulin, is marketed in France for the prevention of alloimmunisation to Rhesus D antigen in obstetrics and after mismatched transfusion. (2) Anti-D human immunoglobulin has been in clinical use for more than 30 years. Its assessment is based more on experience than on well-designed comparative trials. (3) A meta-analysis of 6 trials involving more than 10,000 women demonstrated the efficacy of anti-D human immunoglobulin as prophylaxis after delivery of a Rhesus D-positive infant to a Rhesus D-negative woman. However, the optimal dose regimen and route of administration remain unclear. (4) Some data favour the use of anti-D human immunoglobulin after abortion, as it appears to reduce immunisation rates from about 3-4% to 0.4%. (5) Anti-D human immunoglobulin is likely to be effective antenatally in circumstances or procedures carrying a risk of maternal exposure to fetal red cells, although this has not been proved in comparative trials. (6) Efficacy of anti-D human immunoglobulin in anti-D alloimmunisation after mismatched transfusions has been described only in small clinical pharmacology studies in healthy volunteers. (7) The manufacturing process does not rule out a theoretical risk of nude virus or prion transmission, although no cases have been reported with NateaD degrees . PMID- 11503860 TI - Methylphenidate and narcolepsy: new indication. When modafinil fails. AB - (1) Narcolepsy (daytime bouts of drowsiness) is sometimes associated with cataplexy, and can be incapacitating. The best-assessed treatment is modafinil, which has no demonstrated efficacy on cataplexy. (2) After many years of off license use in the treatment of narcolepsy, methylphenidate is now licensed for this indication in France. (3) According to the results of our literature search, which includes a clinical expert report from Novartis Pharma, the file on methylphenidate in narcolepsy is mainly based on "clinical experience": only three case series, totalling fewer than 200 patients, have been published. (4) These non comparative studies suggest an effect of high dose methylphenidate on daytime drowsiness and cataplexy. (5) The adverse effects of methylphenidate are those of amphetamine psychostimulants, i.e. mainly neuropsychological disorders, cardiovascular effects, loss of appetite and a limited risk of excessive use by some patients. PMID- 11503861 TI - Repaglinide and diabetes: new preparation. No better than glucose-lowering sulphonamides. AB - (1) In France, repaglinide is indicated for the treatment of type 2 diabetes, alone or in combination with metformin, when lifestyle measures are inadequate. (2) The clinical file on repaglinide contains no trials based on morbidity and mortality end points. (3) According to comparative trials versus glucose lowering sulphonamides, the glucose-lowering effect of repaglinide seems to be equivalent to that of glibenclamide. (4) The adverse effect profile of repaglinide appears similar to that of glucose-lowering sulphonamides. Hypoglycaemia is the most frequent adverse effect. In comparative trials, hypoglycaemia was as frequent in patients on repaglinide as in those on glucose lowering sulphonamides. (5) Drug interactions can occur between repaglinide and some enzyme inhibitors or inducers. (6) Treatment with repaglinide at the maximal dose recommended in France is nearly 4 times as costly as with glibenclamide at the maximal dose. PMID- 11503862 TI - Civilian Health and Medical Program of the Uniformed Services (CHAMPUS)/TRICARE; partial implementation of Pharmacy Benefits Program; implementation of National Defense Authorization Act Medical Benefits for fiscal year 2001. Office of the Secretary, DoD. Interim final rule. AB - This interim final rule implements several sections of the Floyd D. Spence National Defense Authorization Act for Fiscal Year 2001. The rule allows coverage of physical examinations for beneficiaries ages 5 through 11 that are required in connection with school enrollment; provides an additional two-year period for survivors of deceased active-duty members to remain eligible for TRICARE medical and dental benefits at active-duty dependent rates; extends eligibility for medical and dental benefits to Medal of Honor recipients and their immediate dependents in the same manner as if the recipient were entitled to retired pay; partially implements the Pharmacy Benefits Program establishing revised copays and cost-shares for the prescription drug benefit; implements the TRICARE Senior Pharmacy Program by establishing a new eligibility for prescription drug benefits for Medicare-eligible retirees; allows a waiver of copayments, cost-shares, and deductibles for all Uniformed Services TRICARE eligible active duty family members residing with their TRICARE Prime Remote eligible Active Duty Service Member Sponsor within a TRICARE Prime Remote designated area until implementation of the TRICARE Prime Remote for Family Member Program or October 30, 2001, whichever is later; provides for the elimination of TRICARE Prime copayments for active duty family members enrolled in TRICARE Prime; provides for the reimbursement of reasonable travel expenses for TRICARE Prime beneficiaries referred by a primary care provider to a specialty care provider who provides services over 100 miles away; and reduces the maximum amount which retirees, their family members and survivors would be liable from $7,500 to $3,000. The Department is publishing this rule as an interim final rule in order to meet statutorily required effective dates. Public comments, however, are invited and will be considered as to possible revisions to this rule. PMID- 11503863 TI - Exemption from the make inoperative prohibition. National Highway Traffic Safety Administration (NHTSA), DOT. Final rule. AB - NHTSA is taking action to facilitate the modification of motor vehicles so that persons with disabilities can drive or ride in them. The agency is accomplishing this by issuing a limited exemption from a statutory provision that prohibits specified types of commercial entities from either removing safety equipment or features installed on motor vehicles pursuant to the Federal motor vehicle safety standards or altering the equipment or features so as to adversely affect their performance. The exemption is limited in that it allows repair businesses to modify only certain types of Federally-required safety equipment and features, under specified circumstances. PMID- 11503864 TI - Clinical chemistry and clinical toxicology devices; classification of B-type natriuretic peptide test system. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the B-type natriuretic peptide (BNP) test system into class II (special controls). The special control that will apply to this device is a guidance document entitled "Class II Special Control Guidance Document for B-Type Natriuretic Peptide Premarket Notifications; Final Guidance for Industry and FDA Reviewers." The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying these devices into class II (special controls) in order to provide a reasonable assurance of the safety and effectiveness of the device. PMID- 11503865 TI - Medical devices; reclassification of the shoulder joint metal/polymer/metal nonconstrained or semi-constrained porous-coated uncemented prosthesis. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is announcing that it is reclassifying the shoulder joint metal/polymer/metal nonconstrained or semi-constrained porous coated uncemented prosthesis intended to replace a shoulder joint from class III to class II (special controls). The agency is also announcing that it has issued an order in the form of a letter to the Orthopedic Surgical Manufacturers Association (OSMA) reclassifying the device. The special control that will apply is a guidance document entitled "Class II Special Controls Guidance: Shoulder Joint Metal/Polymer/Metal Nonconstrained or Semi-Constrained Porous-Coated Uncemented Prosthesis." The agency is classifying this device into class II because special controls, in addition to general controls, would provide reasonable assurance of the safety and effectiveness of the device, and there is sufficient information to establish special controls. PMID- 11503866 TI - Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); TRICARE Dental Program. Office of the Secretary, DoD. Final rule. AB - On October 23, 2000 (65 FR 63202), the Department of Defense published a final rule on TRICARE Family Member Dental Plan. The rule had an effective date that began during the Presidential Moratium on Rules, therefore, this rule is republished to change the effective date to April 1, 2001. This rule is published exactly as previously published. No changes have been made. It revises the comprehensive CHAMPUS regulation pertaining to the Expanded Active Duty Dependents Benefit Plan, or more commonly referred to as the TRICARE Family Member Dental Plan (TFMDP). The TFMDP limited eligibility to eligible dependents of active duty members (under a call or order that does not specify a period of thirty (30) day or less). Concurrent with the timeframe of the publication of the proposed rule, the Defense Authorization Act for Fiscal Year 2000 (Pub. L. 106 65, sec. 711) was signed into law and its provisions have been incorporated into this final rule. The Act authorized a new plan, titled the TRICARE dental program (TDP), which allows the Secretary of Defense to offer a comprehensive premium based indemnity dental insurance coverage plan to eligible dependents of active duty members (under a call or order that does not specify a period of thirty (30) days or less), eligible dependents of members of the Selected Reserve and Individual Ready Reserve, and eligible members of the Selected Reserve and Individual Ready Reserve. The Act also struck section 1076b (Selected Reserve dental insurance), or Chapter 55 of title 10, United States Code, since the affected population and the authority for that particular dental insurance plan has been incorporated in 10 U.S.C. 1076a. Consistent with the proposed rule and the provisions of the Defense Authorization Act for Fiscal Year 2000, the final rule places the responsibility for TDP enrollment and a large portion of the appeals program on the dental plan contractor; allows the dental plan contractor to bill beneficiaries for plan premiums in certain circumstances; reduces the former TFMDP enrollment period from twenty-four (24) to twelve (12) months; excludes Reserve component members ordered to active duty in support of a contingency operation from the mandatory twelve (12) month enrollment; clarifies dental plan requirements for different beneficiary populations; simplifies enrollment types and exceptions; reduces cost-shares for certain enlisted grades; adds anesthesia as a covered benefit; provides clarification on the Department-s use of the Congressional waiver for surviving dependents; incorporates legislative authority for calculating the method by which premiums may be raised and allowing premium reductions for certain enlisted grades; and reduces administrative burden by reducing redundant language, referencing language appearing in other CFR sections and removing language more appropriate to the actual contract. These improvements will provide Uniformed Service members and families with numerous quality of life benefits that will improve participation in the plan, significantly reduce enrollment errors and positively effect utilization of this important dental plan. The proposed rule was titled the - TRICARE Family Member Dental Plan. PMID- 11503867 TI - Finance issue brief: insurance reform. AB - Individual and small group market reforms are intended to increase access to health insurance. The majority of the working uninsured are employed by small businesses, prompting state legislatures to make changes in the small group market. Many states did this on their own; however, to ensure that health insurance was reaching the most people possible, the federal government enacted the Health Insurance Portability and Accountability Act (HIPAA) of 1996. This act required states without extensive law in the small group and individual insurance market to enact laws to comply with provisions such as guaranteed issue, guaranteed renewal, preexisting conditions, portability and nondiscrimination. It also delved into other areas, including long-term care, inpatient care after childbirth, medical savings accounts and mental health parity. PMID- 11503868 TI - Organizing for a new world of health care. AB - At press time, Congress and President Clinton were negotiating how many billions of dollars to cut from Medicaid, and how to structure future federal health care contributions to the states. Regardless of the outcome of those talks, Medicaid cutbacks are only one of several important threats to the health of the poor. In this issue of States of Health, advocates talk about useful policies, tactics, and organizing strategies for maintaining an acceptable level of health care for vulnerable people. PMID- 11503869 TI - Making the Medicaid-welfare disconnect work for low-income families. AB - Medicaid has historically functioned as the health insurance program for families on welfare, but the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 (PRWORA) "disconnected" the two programs. The idea was that families should not lose Medicaid just because they left welfare under PRWORA's new work requirements and time limits. However, de-linkage has not worked as Congress intended. The states have failed to implement the disconnect properly, causing hundreds of thousands of poor families to drop off the medicaid rolls; most are now uninsured. This issue of States of Health looks at the Medicaid-welfare disconnect and how advocates are responding to protect families and children. PMID- 11503870 TI - Consolidation in the health care market: good or bad for consumers. AB - The health care system has been transforming itself; the big fish are eating the small fish; the small fish are combining to grow big. Providers, including hospitals, physicians, and health plans, are aggressively buying, selling, merging, entering into joint ventures, and otherwise consolidating into larger affiliations. These new enterprises, in turn, form networks that can deliver all the health services a patient may need in a lifetime--acute, home health, and nursing home care, and more--and then compete for insurance contracts to cover the greatest possible number of people. While this market reorganization seems to have a role in tempering spiraling costs, it raises other issues of choice, quality, and access. This issue of States of Health examines how consumer interests are faring in the changing market place. PMID- 11503871 TI - Data and information for evaluating managed care. AB - Despite the rapid spread of managed care, its impact on quality and availability of health care services is not yet clear. That's because, in general, health plans are failing to collect, and regulators failing to require, the kind of data that could be useful. This issue of States of Health explores what information we need to collect, analyze, and disclose in order to assess managed care. PMID- 11503872 TI - Monitoring the impacts of the changing health care system. AB - Although the rapidly changing health care delivery system requires careful watching, information that could warn of problems with the quality or availability of care often doesn't exist. This makes the advocate's role in monitoring how consumers are faring more important than ever. In this issue of States of health, advocates share their experiences concerning what situations to monitor, methods for gathering information, and how to use the results to push for improvements. PMID- 11503873 TI - Health care philanthropies: how communities can participate. AB - When a nonprofit hospital or health plan converts to for-profit status, the value of its assets endows a charitable foundation. As a result, billions of health care dollars are being shifted into new philanthropic institutions with an explicit mission to "improve the health of the community." But this issue of States of Health argues that mission can only be accomplished if consumers are involved significantly in the conversion process. PMID- 11503874 TI - Regulating risks: consumer protections in provider health networks. AB - As competition for health care dollars grows fierce, physicians and hospitals are going head to head with insurance companies. They are forming new partnerships, broadly called provider-sponsored organizations, that cut out the middleman and have the potential to furnish better care at lower prices. But legislators, regulators, and consumers must understand the dangers inherent in the structure of some PSOs. PMID- 11503875 TI - Consumer advocacy in Medicare HMOs. AB - Rising out-of-pocket health care costs and premiums for Medicare supplemental insurance are driving many beneficiaries out of traditional fee-for-service Medicare and into health maintenance organizations. These consumers give up unrestricted provider choice in exchange for controlled costs and some additional service. However, in the context of weak oversight by the federal Health Care Financing Administration, the push by health plans to increase profits has meant that vulnerable Medicare HMO enrollees may not receive the services and consumer protections required by law. PMID- 11503876 TI - The Ohio Blue Cross conversion: building an opposition. AB - With good reason, there has been loud public opposition to the proposed sale of Blue Cross of Ohio to Columbia/HCA Healthcare Corporation. If approved, the deal would allow the hospital giant also to control insurance coverage for one and a half million subscribers. Many aspects of the deal are alarming, but foremost is that assets belonging to the public would be sold to a private corporation. As States of Health goes to print, the Department of Insurance decision whether to approve the sale has not been released, though news stories indicate the answer is no. Regardless of the decision, consumers won: without their intervention, regulators were expected to give the deal pro forma approval. Instead, the public was quickly educated, and the deal received the scrutiny it deserves. PMID- 11503877 TI - Preserving and expanding services when hospitals change hands. AB - An estimated 700 hospitals were merged, sold, or otherwise changed hands in 1996. When this happens, communities worry that needed services will be lost. But each transaction is an opportunity to reexamine hospital services and push for improvement. This issue of States of Health shows how consumer groups around the country are using market transactions to advance services and expand community participation. PMID- 11503878 TI - Community benefits in a changing health care market. AB - Market changes in the health industry--mergers, acquisitions, and other transactions--are eliminating many of the traditional sources of care for people who have no insurance or poor coverage. There are fewer public or private nonprofit hospitals with a charitable mission. Moreover, through Medicaid contracting, a portion of the funds that once supported broad public health goals now go to private HMOs that serve only their own members. Advocates are responding with the demand that health providers--nonprofit and for-profit, hospitals and health plans--collaborate with the residents of communities where they do business to improve people's health. PMID- 11503879 TI - The Ohio Blue Cross conversion: building an opposition. AB - In March, state insurance regulators rejected the bid of Blue Cross of Ohio to sell the business to Columbia/HCA, saying it would not be in the public interest. The decision came as a shock to those who assumed the transaction would get pro forma approval. The fact that it received press, public, and regulatory attention is due largely to the efforts of a coalition of consumer interests, supported by Community Catalyst. This issue of States of Health outlines how Community Catalyst and other groups successfully focused and strengthened opposition to a deal that disposed of public assets without public participation or representation. PMID- 11503880 TI - State and federal insurance reforms for individuals and small groups. AB - Forty-six states and the federal government have enacted legislation to eliminate some of the worst forms of discrimination by health insurance companies against individuals and people who buy in small groups. This issue of States of Health examines what the federal Health Insurance Portability and Accountability Act will--and won't--do for vulnerable people. It highlights the continuing need for state reforms and looks at a role for advocates. PMID- 11503881 TI - Expanding health insurance for children. AB - This issue of States of Health provides information that advocates, consumers, legislators, and other policymakers can use in crafting effective state health insurance programs for children. It looks at states' options under the rules and funding of the State Children's Health Insurance Program, the 10-year, $50 billion federal package enacted in 1997 as Title XXI, highlighting the critical program design issues that may mean the difference between healthy children and wasted opportunity. PMID- 11503882 TI - Purchasing cooperatives for small employers. AB - Despite a booming economy, the number of uninsured Americans is rising. It hit nearly 42 million in 1996. Many of the uninsured work at businesses with fewer than 50 employees. Because small firms have traditionally found it difficult to provide health benefits, purchasing cooperatives have grown in scope and size across the country in recent years. By bringing small businesses together to buy insurance as a group, these organizations can help employers provide greater choice to their workers at a lower cost. However, to operate well in the insurance market, purchasing cooperatives must be well-designed and provided with adequate legal protections. PMID- 11503883 TI - Making managed care work for people with disabilities. AB - Over two years ago, with states increasingly moving to contract with managed care organizations for the care of people with disabilities and chronic illnesses, States of Health explored the concerns of some of these health care consumers and their families. Worries centered on the health plans' lack of experience serving disabled and chronically ill patients; the prevalence of a medical model of care, which undervalues services that enable a person to maintain a high quality of daily life; and incentives to cut costs that would result in inappropriate or negligent care. Little has been resolved since then. A few health plans--mostly small specialty plans--serve as oases of best practices, but their attitudes and ways of operating are far from adoption across the board. Advocates argue that the best targeted, most reliable consumer protections involve consumers themselves in the planning, design, and implementation of managed care programs. PMID- 11503884 TI - Overcoming problems with dental care for Medicaid beneficiaries. AB - Lack of access to dental care means more than dirty teeth to many vulnerable consumers. It also is related to pain, poor nutrition, and the inability to accomplish tasks of daily living. This issue of States of Health examines the nature of the problem and the variety of ways advocates are addressing it. PMID- 11503885 TI - "Crowd-out": what it means for children's health insurance. AB - As states finalize proposals for the federal Title XXI Children's Health Insurance Program (CHIP), consumers, advocates, legislators, and policymakers across the country are facing the issue of "crowd-out." How can they prevent CHIP's new public funds from "crowding out"--or supplanting--private funds now used to insure children? This issue of States of Health looks at research on crowd-out and reviews what some states have learned as a result of previous Medicaid expansions. Their experiences shed light on the challenge of making sure that Title XXI health care dollars reach the intended consumers, children who lack adequate insurance. PMID- 11503886 TI - The Columbia/HCA hospital sales: an opportunity to re-focus on community benefits. AB - Columbia/HCA, currently the target of federal fraud investigations that have led to indictments of four top executives, announced plans in late 1997 to sell 115 hospital facilities. In a period of frenetic mergers, this sale will most affect communities in which Columbia has put multiple facilities on the auction block, yet this round of restructuring also foreshadows an increasingly consolidated- and profit-driven--industry. States of Health explores how community intervention in these sales may offer a chance to see that health needs are not further sacrificed to industry profits. PMID- 11503887 TI - Strengthening the free care safety net. AB - For many of the estimated 43 million people in this country who have no health insurance, free care is often the only health care available. With competition forcing hospitals to cut costs, consumer advocates face new challenges as they seek to ensure that free care remains available to all who need it. This issue of States of Health explores the problems facing both consumers who rely on free care and those who provide that care. It highlights what some advocates and communities are doing to address those concerns. PMID- 11503888 TI - The new health philanthropy: ensuring the effective use of conversion foundation assets. AB - Across the country, the conversion of nonprofit hospitals and health insurance plans to for-profit status is resulting in the creation of new philanthropic foundations. The preservation of these assets--and their continued use for public benefit--is required by law. This issue of States of Health discusses an additional challenge facing consumers and communities: ensuring that these assets are most effectively used to promote community health. PMID- 11503889 TI - Medicare HMO marketing and information: keeping the focus on what consumers need. AB - Across the country, Medicare beneficiaries face complicated choices about health insurance and HMOs, but too often they lack the information they need to make good decisions. Instead, they rely on marketing information that is frequently inadequate, sometimes downright misleading. This issue of States of Health looks at efforts to improve the ways that HMOs market their Medicare plans and assesses whether "choice" really benefits beneficiaries. PMID- 11503890 TI - Unfinished business: the restoration of immigrant health access after welfare reform. AB - One in ten Americans is a recent immigrant and, until 1996, even the poorest had access to health care through Medicaid. That changed radically with the 1996 Personal Responsibility and Work Opportunity Reconciliation Act (PRWORA), the federal welfare reform law, which sharply restricted immigrant access to public benefits, including Medicaid. This issue of States of Health looks at PRWORA's impact--and what advocates and immigrants are doing to restore health care access for these newcomers, who are among the most vulnerable in our communities. PMID- 11503891 TI - Children's health insurance programs: strategies for outreach and enrollment. AB - In tandem with Medicaid, the State Children's Health Insurance Program (CHIP) could cover most of the 11 million uninsured children in the United States. Nearly half of children without insurance are eligible for existing Medicaid programs, and an estimated 4 million more will be eligible under new CHIP rules. The key to moving toward universal coverage for children is outreach--making sure that every child who qualifies for the programs gains access to them. This issue of States of Health discusses some of the most effective strategies for achieving that goal. PMID- 11503892 TI - Coordination, choice, and value in long-term care. AB - Systemic problems in the nation's long-term care system result in high-cost, low quality care in most states. Consumers must exhaust their savings to become eligible for long-term care coverage through Medicaid, get lost in a maze of fragmented systems funded by different sources, and often are institutionalized unnecessarily. Several states have developed new models to provide long-term care that is better coordinated and includes more social supports. As these examples show, it pays to invest in home and community-based care systems and to help those systems connect to medically based networks. PMID- 11503893 TI - Expanding insurance coverage and access for low-income people. AB - Medicaid provides health insurance for 40 million low-income women and children, elderly, blind, and disabled people. In 1996, state initiatives covered nearly a million more people who have very low incomes but aren't eligible for Medicaid. This issue of States of Health looks at the extent to which such programs to expand insurance coverage actually improve access to care. How many of those who are eligible are enrolled? How many receive appropriate care? And how can we increase those numbers to the limit? PMID- 11503894 TI - Rural issues in managed care. AB - Residents of rural areas are sicker, older, and poorer than the rest of the population. Though their use of health care is relatively low, their need is greater. The ongoing reorganization of the health care financing and delivery systems may have more impact--positive or negative--on rural people than on anyone else. This issue of States of Health examines how to preserve or improve quality and access for rural consumers in managed care. PMID- 11503895 TI - ACOG Committee Opinion no. 257: Committee on Genetics. Genetic evaluation of stillbirths and neonatal deaths. PMID- 11503896 TI - Host immune responses in the course of bovine leukemia virus infection. AB - Bovine leukemia virus (BLV) is a type C retrovirus infecting bovine B cells and causing enzootic bovine leukosis. Since it takes long periods to develop the disease, it is believed that BLV and host immune responses are closely related. In this review, the accumulated data showing close relationship between BLV and host immune responses are summarized in 4 sections. First, we discuss the role of cell-mediated immunity in protecting hosts from BLV infection. Second, several reports showing the relationship between the disease progression and the change of cytokine profiles are summarized. In the third section, we have focused on tumor necrosis factor alpha (TNFalpha) and its two types of receptors, and the possible involvement of TNFalpha in the BLV-induced leukemogenesis is discussed. The expression of TNFalpha has been shown to be regulated by major histocompatibility complex (MHC) haplotype. The resistance to BLV infection is supposed to be established by some innate factors, which are closely related to MHC haplotype. Finally, we propose that a breeding strategy based on the MHC haplotype could be a good approach to control BLV infection. This review includes some recent data from us and other groups. PMID- 11503897 TI - Higher sensitivity in induction of apoptosis in fibroblast cell lines derived from LEC strain rats to ultraviolet B radiation. AB - When lung fibroblast cell lines from LEC and WKAH rats were irradiated with ultraviolet B (UVB) and assayed for colony formation, LEC rat cells showed a higher sensitivity than did WKAH rat cells. The LEC rat cells were approximately 1.5-fold more sensitive to UVB radiation than were the WKAH rat cells in terms of D37 values, which are the doses of UVB required to reduce cell survival to 37%. When the rat cells were irradiated with UVB in the presence of 0.5 M dimethyl sulfoxide (DMSO), which efficiently scavenges free radicals such as hydroxyl radicals, no significant difference was observed between the survival curves of either LEC or WKAH rat cells irradiated with UVB in the presence of 0.5 M DMSO and those irradiated with UVB in the absence of DMSO. Therefore, formation of free radicals may not be involved in cell death induced by UVB radiation. Flow cytometry showed that the percentage of apoptotic cells in the LEC rat cell population increased with post-incubation time after UVB radiation. The proportion of apoptotic cells in the UVB-irradiated LEC rat cell population increased as the dose of UVB was increased. In contrast, no significant proportion of apoptotic cells was observed in the UVB-irradiated WKAH rat cell population. These results showed a higher sensitivity in induction of apoptosis by UVB radiation in LEC rat cells than in WKAH rat cells. PMID- 11503898 TI - Establishment of the prediction table of parturition day with ultrasonography in small pet dogs. AB - To establish a prediction table of parturition day the real-time B-mode ultrasonographic examinations were performed in the 8 pregnant Malteses and 10 Yorkshire terriers (total pups, 25 and 38 pups, respectively) from 18 days of gestation until the parturition. Ovulation was designated the first day of gestation (day 0). Extra fetal and fetal structures were measured from all conceptues. The parameters that exhibited the best correlation to parturition were used to compile a prediction table of parturition day. To testify the precision of the prediction table of parturition day, the 15 pregnant Malteses (48 pups) and 13 pregnant Yorkshire terriers (42 pups) with unknown mating time were examined using ultrasonography. Inner chorionic cavity diameter on days 18 to 37 and fetal head diameter on day 38 to parturition that showed the best correlation to gestational age were the most pertinent to the estimation of gestational age and the prediction of parturition day. The two parameters were used to compile a prediction table of parturition with averaged regression equations. In verificational examinations, with the exception of I Yorkshire terrier (3.6%) having 1 fetus, 18 of 28 bitches (64.3%) delivered exactly on the date predicted and 9 of 28 bitches (32.1%) delivered within I day of the date predicted. Therefore, the prediction table of parturition day seems to be a useful tool of the prediction of parturition day in practice. PMID- 11503899 TI - Genotypes of pestivirus RNA detected in live virus vaccines for human use. AB - Live virus vaccines for human use, 29 monovalent vaccines against measles, mumps, rubella or polio, eight polyvalent vaccines against measles-mumps-rubella and one bacterial polyvalent vaccine against Streptococcus pneumoniae, were tested by reverse transcriptase-nested PCR for the presence of petivirus or pestivirus RNA. Twenty-four samples were selected from European manufacturers, ten were from U.S.A. and four from Japan. Five (13.1%) out of 38 tested samples were positive for pestivirus RNA. Three vaccines (rubella and two measles) were from Europe and two (mumps and rubella) from Japan. The 5'-untranslated genomic region of the contaminant pestivirus RNA were amplified by reverse transcription-PCR and sequenced. Analyses based on primary nucleotide sequence homology and on secondary structures, characteristic to genotypes, revealed that the cDNA sequences belonged to bovine viral diarrhea virus (BVDV). A cDNA sequence, detected from one measles sample, belonged to BVDV-1b genotype. Pestiviral cDNA detected from the Japanese mumps and rubella vaccine samples, belonged to the BVDV genotypes 1a and 1c, respectively. Analysis on two cDNA sequences detected from measles and rubella vaccine samples from Europe showed their appurtenance to a new genotype, BVDV-1d. These findings indicate that contamination by animal pestivirus may occur in biological products for human use. PMID- 11503900 TI - Comparative study of the reproductive organs of Fasciola groups by optical microscope. AB - Reproductive organs of stained and mounted whole specimens of different types of Fasciola (F. hepatica, F. gigantica, and parthenogenetic diploid and triploid flukes) were observed to clarify the structure of their reproductive organs. The results are as follows; 1. Basic structure differences could not be identified. 2. The flukes without sperm, or those with an extremely small quantity in the seminal vesicle. are parthenogenetic Fasciola sp. 3. It was newly discovered that the surface of the cirrus is surrounded by many shallow gutters, and that spines form a line in the gutters. 4. The structure of the reproductive organ on the genus Fasciola are shown in detail in the figures. PMID- 11503901 TI - In vitro antibody response to the tetrapeptide repeats of Plasmodium falciparum circumsporozoite protein by splenocytes from mice infected with P. yoelii yoelii. AB - The antibody response to the recombinant protein, R32tet32, which contained the repetitive sequence (NANP)n of Plasmodium falciparum CSP was determined in C57BL/6 mice during the course of nonlethal infection with Plasmodium yoelii 17X. Marked suppression of the IgG antibody response to R32tet32 occurred when mice were immunized at peak parasitemia (on day 16). In vitro antibody responses of spleen cells from acutely infected mice to R32tet32 were similarly suppressed. Stimulation of normal spleen cells cultured for 5 days with 100 ng/ml of R32tet32 gave an optimal IgG antibody response, but spleen cells from infected mice obtained at peak parasitemia failed to respond to a broad range of antigen concentrations. Cocultivation studies employing enriched lymphocyte populations from infected and uninfected C57BL/6 mice indicated that both T and B cells from infected mice were defective in their response to R32tet32. The response to the repetitive region was restored by the addition of recombinant mouse interleukin-2 (IL-2) at a dose of 50 U/ml to cultures of spleen cells from infected mice. PMID- 11503902 TI - Characterization of classical swine fever virus associated with defective interfering particles containing a cytopathogenic subgenomic RNA isolated from wild boar. AB - Classical swine fever virus (CSFV) strain WB82, isolated from a wild boar in 1982, induced a distinct cytopathic effect (CPE) in primary swine testicle cell culture and in most of the porcine cell lines. This strain of CSFV was found to be composed of two biotypes. cytopathogenic (cp) CSFV, as a minor population, and noncytopathogenic (noncp) CSFV, as a major population. The noncp CSFV (designated strain WB82/E+) was obtained by biological cloning, and it showed the exaltation of Newcastle disease virus phenomenon. In Northern blot analysis and RT-PCR assay, CSFV RNA with a subgenomic (sg) length was detected in addition to full length viral RNA only in the cells in which a CPE had been revealed. These RNAs represent the genomes of typical defective interfering (DI) particles because of the strict dependence on a complementing helper virus and interference with replication of the helper virus. The sg RNA, which exhibits the genomes of the DI particles, lacked the nucleotides of the viral genomic region from Npro to NS2 (4764 bases). When extracted sg RNA was transfected to the cells infected with the WB82/E+ strain, a distinct CPE was observed. Interestingly, the CPE was observed in cells infected with other heterologous noncp CSFV ALD and GPE- strains by sg RNA transfection. The results suggested that these noncp CSFVs act as helper viruses for the replication of sg RNA (DI particles). It was also shown that the cytopathogenicity of strain WB82 is caused by apoptosis. PMID- 11503903 TI - Immunohistochemical studies on the differential maturation of three types of olfactory organs in the rats. AB - Differential maturation of three types of olfactory organs, the olfactory epithelium (OE), the vomeronasal organ (VNO) and the septal olfactory organ of Masera (MO), was examined immunohistochemically in embryonic and newborn rats by the use of antiprotein gene product 9.5 (PGP 9.5) serum. These olfactory organs were derived in common from the olfactory placode as neuroepithelia. In the OE, PGP 9.5-immunopositive olfactory cells first appeared at 13 days of gestation. The OE maturated completely, and showed the same cytological features as in the adult at 20 days of gestation. The MO first appeared as a dense mass of PGP 9.5 immunopositive sensory cells on the most ventrocaudal part of the nasal septum at 15 days of gestation and was evidently isolated from the OE by the decrease of immunopositive cells in the intercalated epithelium between the OE and the MO at 20 days of gestation. However, even at 7 days after birth, the MO did not complete its development and contained sensory cells aggregating in the mass. The VNO was separated from the nasal cavity at 13 days of gestation as a tubular structure of a neuroepithelium including PGP 9.5-immunopositive sensory cells. These cells gradually increased in number in the sensory epithelium of the VNO and extended their dendritic processes to the free surface at 7 days after birth. These findings clarified the differential maturation of these olfactory organs. That is, the OE completes its development before birth, while the MO and VNO after birth. PMID- 11503904 TI - Mutagenicity of water samples from five cities in Korea. AB - Four doses (equivalent to 4, 2, 1, and 0.5 liter water) of organic extracts from raw, treated and drinking waters sampled from seven different treatment plants in five cities in Korea were challenged to the Ames test using S. typhimurium strains TA98 and TA100 in the presence/absence of S9 mix. The mutagenicity was usually observed from chlorine-treated (28.6%) and drinking (42.9%) waters rather than raw (3.4%) waters. The strain TA98 (33.3%) was more sensitive to detect the mutagenicity of water samples than the strain TA100 (16.7%). However, the absence of S9 mix showed higher mutagenic activity of waters compared to the presence of S9 mix, corresponding to the detection of 42.9% and 7.1%, respectively. These results indicate that the bacterial mutagenicity of treated and drinking waters may be derived from chlorination in water treatment plants but that the mutagenicity in humans may be limited due to enzymatic metabolism. PMID- 11503905 TI - Cycle of the seminiferous epithelium in the Java fruit bat (Pteropus vampyrus) and the Japanese lesser horseshoe bat (Rhinolophus cornutus). AB - The cycle of the seminiferous epithelium in the Java fruit bat, Pteropus vampyrus, and the Japanese lesser horseshoe bat, Rhinolophus cornutus, was investigated by light microscopy and the characteristics of spermiogenesis were compared between these two species. In the Java fruit bat, the cycle of the seminiferous epithelium was divided into 11 stages and developing spermatids were subdivided into 13 steps. While in the Japanese lesser horseshoe bat, the cycle of the seminiferous epithelium was divided into 10 stages and developing spermatids were subdivided into 13 steps. Excepting slight morphological differences, the characteristics of acrosomal formation in both species were almost similar with each other. In the Java fruit bat after stage VII, the acrosome gradually elongated, flattened and finally became scoop-like in shape. In the Japanese lesser horseshoe bat after stage VIII, the acrosome elongated, flattened and then slightly shortened. Before spermiation, the acrosome became long spatula-like in shape. The elongation and flattening of spermatids in these two species were similar to those in insectivores. The finding may reflect the fact that the order Chiroptera is phylogenetically close to the order Insectivora. PMID- 11503906 TI - Localization of proliferative and apoptotic cells in the kidneys of ICR-derived glomerulonephritis (ICGN) mice. AB - The ICR-derived glomerulonephritis (ICGN) mouse is a novel inbred mouse strain with a hereditary nephrotic syndrome, considered to be a good model of human idiopathic nephrotic syndrome and develops proteinuria, hypoproteinemia and anemia. In the present study, we compared the cell kinetics in the kidneys of ICGN mice with age-matched ICR mice as normal controls. The proliferating cells were visualized by 5-bromo-2'-deoxyuridine labeling, and apoptotic cells were determined by terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling. Many proliferating epithelial cells of renal tubules, glomerular mesangial cells and tublointerstitial fibroblast like cells were observed in the kidneys of ICGN mice, but no proliferating cells were seen in the kidneys of ICR mice. Apoptotic cells had round nuclei, and were observed only in the tubulointerstitium in the kidneys of ICGN mice but not in that of controls. The proliferation of renal tubular epithelial cells may represent a compensatory response, and that of mesangial and fibroblast-like cells may play a pathogenic role in nephrotic syndrome. Apoptosis in tubulointerstitial cells with round nuclei may have been erythropoietin-producing cells, and probably caused anemia. PMID- 11503907 TI - The formation of apical delta of the permanent teeth in dogs. AB - To determine the process of formation of apical delta, a histological study on the permanent teeth was carried out in dogs. A litter of 7 clinically healthy beagle dogs and 33 adult dogs (4- to 15- year-old) of 12 breeds with periodontal disease were used for the experiments. Teeth extracted from 6-,7-,8- and 9-month old beagles were sectioned and stained with HE solution. Tooth roots obtained from adult dogs with periodontal disease were ground. Each tooth was classified into the following root types under a light microscope: Type I (no apical delta = no apical closure), II (few apical delta), IIIA (low apical delta) and IIIB (high apical delta). In the 6-month-old beagles, more than half the tooth roots were classified as type I. In the 7-month-old beagles, type IIIB apical delta was the most predominant and types I, II and IIIA apical delta were occassionally seen. Apical closure and delta were observed in all beagles at 8 months of age histologically. In the 8- and 9-month-old beagles, all root apexes observed were type IIIB. Most of the 314 tooth roots extracted from 33 adult dogs were type IIIB, but a few were type IIIA. PMID- 11503908 TI - Role of reticulocytes on gametocytogenesis in chickens infected with Leucocytozoon caulleryi. AB - Reticulocytes, known as late polychromatic erythrocytes, were induced in blood of chickens infected with Leucocytozoon caulleryi by bleeding when the second generation merozoites were released into the blood from the second-generation schizonts. The second-generation merozoites preferentially invaded into reticulocytes and developed to stage II gametocytes. Enhanced development of stage II gametocytes to mature gametocytes was observed in the reticulocytes in vivo and in vitro in the bleeding group. Nevertheless, invasion of reticulocytes by the second-generation merozoites was not considered to be absolutely necessary for the development of gametocytes. PMID- 11503909 TI - Molecular properties of the matrixprotein(M) gene of the lapinized rinderpest virus. AB - The nucleotide sequence of the matrixprotein (M) gene of the lapinized rinderpest virus (RPV-L) was determined. The full-length cDNA of the RPV-L M gene is composed of 1460 base pairs and is supposed to contain an open reading frame of 1005 nucleotides encoding on M protein of 335 amino acids. The homology of the predicted amino acid among congeneric morbilliviruses such as RPV Kabete 'O' strain (wild strain of RPV), RPV RBOK strain (vaccine strain of RPV for cattle), measles virus (MV), and canine distemper virus (CDV), is approximately 94%, 93%, 87% and 77%, respectively. In the present study, all coding regions of the RPV-L strain have been determined. PMID- 11503910 TI - Activation of GABA(A) receptors in the accessory olfactory bulb does not prevent the formation of an olfactory memory in mice. AB - When female mice are mated, they form a memory to the pheromonal signal of their male partner. The neural mechanisms underlying this memory involve changes at the reciprocal dendrodendritic synapses between glutamatergic mitral cells and gamma aminobutyric acid (GABA)-ergic granule cells in the accessory olfactory bulb (AOB). Blockade of GABA(A) receptors in the AOB leads to the formation of an olfactory memory. In an attempt to disrupt memory formation at mating, we used local infusions of the GABA(A) receptor agonist muscimol into the AOB during the critical period for memory formation. Muscimol across a wide range of doses (1 1000 pmol) did not prevent memory formation. The resistance of this memory to GABA(A) receptor activation may reflect the complexity of synaptic microcircuits in the AOB. PMID- 11503911 TI - Mast cell MMP-9 production enhanced by bacterial lipopolysaccharide. AB - Although mast cells contribute to host protective immunity against bacterial infections, the exact mechanism of their recruitment at the affected site has been unclear. Recently, we have reported that both mouse and human mast cells are capable of producing matrix metalloproteinase (MMP)-9, a matrix-degrading enzyme necessary for leukocyte transmigration. Here, we demonstrated that bacterial lipopolysaccharide (LPS) enhanced MMP-9 production of mouse bone marrow derived cultured mast cells. This action of LPS was partially suppressed by the pretreatment of cultured mast cells with a protein kinase C (PKC) inhibitor, indicating the possible involvement of PKC signaling pathways in the production of MMP-9 by LPS. Thus, these suggest the upregulation of mast cell MMP-9 by bacterial components, thereby resulting in their migration at the affected site. PMID- 11503912 TI - Detection of ehrlichial infection by PCR in dogs from Yamaguchi and Okinawa Prefectures, Japan. AB - Species-specific nested polymerase chain reaction (PCR) was used to detect the presence of possible canine ehrlichial agents (Ehrlichia canis, E. chaffeensis, E. ewingii, E. equi and E. platys) and monocytic ehrlichial agents found in Japan (E. muris and a recently discovered Ehrlichia species detected from Ixodes ovatus) in blood samples from dogs in Yamaguchi and Okinawa Prefecture, Japan. Partial sequence of E. platys was detected from 1 of 67 dogs (1.5%) tested from Yamaguchi Prefecture and 24 out of 87 (27.6%) in the subtropical Okinawa Prefecture. Dogs in Okinawa and Miyako Islands had a higher positive rate (69.2 and 45.0%, respectively) than Ishigaki Island (11.1%). Another dog in Yamaguchi Prefecture had a positive PCR reaction to the Ehrlichia sp. detected from I. ovatus. No other Ehrlichia were found in these samples. PMID- 11503913 TI - Aberrant branch of the bronchoesophageal artery resembling patent ductus arteriosus in a dog. AB - An anomalous shunt between the bronchoesophageal artery and pulmonary artery was diagnosed in a 1-year- old, 3.5 kg female Miniature Dachshund by selective contrast angiography. A cardiac murmur had been observed in the dog during examination at another hospital. The machinery murmur was auscultated at the left side of the base of the heart. Although thoracic radiography revealed mild cardiomegaly, the characteristic findings of patent ductus arteriosus (PDA), including as aortic arch enlargement and pulmonary artery enlargement were not observed. Echocardiography demonstrated shunting of blood flow presumably from the arterial duct at the pulmonary artery carina. Based on the above findings the case was diagnosed as PDA. Angiocardiography was performed to confirm the diagnosis in preparation for surgical treatment, but later we confirmed that the shunt vessel was not PDA, but apparently a branch of the bronchoesophageal artery. The shunt vessel was branching in a complicated manner and shunted to the pulmonary artery. PMID- 11503914 TI - The first host record of trichinosis in a red fox, Vulpes vulpes japonica, from Aomori Prefecture, northern Honshu, Japan. AB - A case of wildlife trichinosis was found in a red fox (Vulpes vulpes japonica) captured at Rokkasho, Aomori Prefecture on November 27, 1998. Trichinella larvae were obtained from almost all of the muscle tissues except for the masseter. The highest number of larvae per gram of tissue was found in the muscles of the gluteal region and throat. The lowest number was found in the diaphragm and tongue. Trichina cysts within the muscle fibers had groups of fatty cells at the poles, and minimal tissue reaction was observed around the cyst. No calcification was found in the cyst. These morphological findings suggested that the considerable time had elapsed since the invasion. This is the first case of trichinosis in a red fox in Japan. PMID- 11503915 TI - Lymphoproliferative responses in pigs infected with Mycobacterium avium. AB - Naturally infected cases of swine mycobacteriosis were divided into two groups, localized infection (LI) and disseminated infection (DI). Lymphoproliferative response (LPR) was then examined to estimate their immunological states. Both control and LI groups showed strong response to Concanavalin A (Con A) and phytohemagglutinin (PHA) in the LPR, and lymphocytes recovered from the LI responded well to purified protein derived from M. avium (PPD). On the other hand, the DI group showed weak response to both Con A and PHA, despite their strong response to PPD stimulation. These data suggest that the low LPR to Con A and PHA observed in the DI groups was probably not due to the general unresponsiveness of T-cells. PMID- 11503916 TI - Imaging the human corpus luteum. PMID- 11503917 TI - Breast ductal carcinoma in situ. PMID- 11503918 TI - Sonographic and Doppler characteristics of the corpus luteum: can they predict pregnancy outcome? AB - OBJECTIVE: To determine whether there is a relationship between gray scale or Doppler characteristics of the corpus luteum and first-trimester pregnancy outcome. METHODS: We conducted a prospective study of patients with spontaneous singleton pregnancies between 5 and 8 weeks' gestation. The corpus luteum size, sonographic appearance, resistive index, and peak systolic velocity were measured on transvaginal sonography. Maternal use of exogeneous progesterone was recorded. Only patients with known first-trimester outcome were included. RESULTS: There were 201 study patients. The corpus luteum could be visualized in 197 (98%) and had a mean +/- SD size of 1.9 +/- 0.6 cm, a mean resistive index of 0.50 +/- 0.08, and a peak systolic velocity of 20.5 +/- 11.2 cm/s. There were 151 first trimester survivors (75.1 %) and 50 spontaneous losses (24.9%). In a comparison of the survivors and losses, there was no significant difference in mean corpus luteum size (1.9 versus 1.7 cm; P = .10, t test), mean resistive index (0.50 versus 0.50; P = .71, t test), peak systolic velocity (21 versus 19 cm/s; P = .29, t test), or sonographic appearance (P = .78, chi2 test). The lack of association between corpus luteum characteristics and outcome persisted when cases were stratified by progesterone use and the presence or absence of a heartbeat on the study sonogram. CONCLUSION: There is no apparent relationship between the characteristics of the corpus luteum and first-trimester pregnancy outcome. PMID- 11503919 TI - Preoperative evaluation of pelvic tumors by Doppler and three-dimensional sonography. AB - OBJECTIVE: To study a spectrum of systems (two-dimensional transvaginal, transvaginal color Doppler, three-dimensional, three-dimensional power Doppler, and contrast-enhanced three-dimensional power Doppler sonography) for preoperative evaluation of pelvic tumors. METHODS: Two hundred ninety-two patients were evaluated by the 5 complementary methods in preoperative sonographic assessments. We examined adnexal and endometrial morphology, thickness, and volume by two- and three-dimensional sonography and analyzed blood flow by transvaginal color, pulsed Doppler, and three-dimensional power Doppler sonography in all examined patients. In 89 patients with complex adnexal lesions of uncertain malignancy, contrast-enhanced three-dimensional power Doppler sonography was performed. RESULTS: Morphologic assessment by three-dimensional sonography yielded additional information in 58% of cases compared with two dimensional sonography. Furthermore, this modality was superior to two dimensional sonography in accurate depiction and diagnosis of 2 cases of fallopian tube carcinoma. Combined morphology and vascular indexing reached sensitivity of 97% and specificity of 99%. Endometrial volume in patients with malignant disease was significantly different (28.2 +/- 0.02 cm3) from that in those who had hyperplasia (7.81 +/- 0.03 cm3), polyps (3.5 +/- 0.02 cm3), or normal endometria (0.8 +/- 0.02 cm3). With combined morphologic and three dimensional power Doppler examination of endometrial lesions, sensitivity and specificity reached 89% and 97%, respectively. CONCLUSIONS: Combined morphologic and vascular imaging improves preoperative assessment of gynecologic tumors. PMID- 11503920 TI - Assessment of a new logistic model in the preoperative evaluation of adnexal masses. AB - OBJECTIVE: To assess a new logistic regression model developed to predict malignancy in adnexal masses. METHODS: In the first part of this study, we developed a logistic model by applying logistic regression analysis in a series of 268 adnexal masses (203 benign and 65 malignant lesions) in 248 patients (mean age, 43.6 years; SD, 14.2 years) evaluated and treated at our institution. Eleven parameters were entered in the logistic regression analysis in a forward stepwise way. In the second part of the study, we evaluated the model's diagnostic performance in a further set of 135 adnexal masses (103 benign and 32 malignant tumors) in 129 patients (mean age, 44.4 years; SD, 14.6 years). This diagnostic performance was compared with that of age, tumor volume, Sassone's and Ferrazzi's B-mode ultrasonographic morphologic scoring systems, serum cancer antigen 125 level, and the tumor's lowest resistive index. Comparison was done by calculating the area under the receiver operating characteristic curve. RESULTS: In logistic analysis, only menopausal status, the presence of papillary projections, the logarithm of the cancer antigen 125 value, tumor blood flow location, and the lowest resistive index were retained in the model. The model had the best area under the curve (0.97), significantly higher than patient age (area under the curve, 0.78; P = .001), tumor volume (area under the curve, 0.68; P < .0001), cancer antigen 125 (area under the curve, 0.88; P = .008), lowest resistive index (area under the curve, 0.85; P = .011), Ferrazzi's scoring system (area under the curve, 0.89; P = .01), and maximal peak systolic velocity (area under the curve, 0.71; P< .0001). Comparison with Sassone's scoring system (area under the curve, 0.91) did not reach statistical significance, but a clear trend was found (P = .116). CONCLUSIONS: The model had the best diagnostic performance for discriminating between benign and malignant adnexal masses. A clinical prospective evaluation is needed to confirm its actual value. PMID- 11503921 TI - Subtorsion of the ovary: sonographic features and clinical management. AB - OBJECTIVE: To define the sonographic imaging criteria of ovarian subtorsion. METHODS: Fourteen women 16 to 35 years of age with suspected unilateral twisted ovaries were sonographically evaluated in a tertiary care center before laparoscopy. Transvaginal sonography was performed to determine the longest ovarian dimension, diameter of the periovarian vessels, and degree of intraovarian blood flow and to evaluate ovarian morphology. Laparoscopy was performed to verify the sonographic findings and to perform detorsion. RESULTS: All ovaries with subtorsion were of normal size, although still significantly longer than the contralateral, normal ovaries (47.5 and 29.6 mm, respectively; P = .001). The mean diameter of the periovarian vessels was significantly greater for the ovaries with subtorsion than the normal ovaries (29.9 and 19.2 mm, respectively; P= .0001). At laparoscopy, the ovary was twisted less than 180 degrees in 9 patients and 180 degrees to 360 degrees in 5. The degree of twisting correlated significantly with the size of the ovary (P = .016) and diameter of the periovarian vessels (P = .046). The 3-month sonographic follow-up after detorsion revealed bilateral normal ovarian dimensions with proper follicular growth. CONCLUSIONS: Congestion of the periovarian vessels with normal ovarian morphology and persistent blood flow on sonography, combined with the presence of appropriate clinical signs, may be indicative of ovarian subtorsion. PMID- 11503922 TI - Significance of the solid component in predicting malignancy in ovarian cystic teratomas: diagnostic considerations. AB - OBJECTIVE: To determine whether gray scale characteristics of the solid components of cystic ovarian teratomas exist that could differentiate more common benign forms from malignant variants. METHODS: We retrospectively reviewed the sonographic images of 188 ovarian teratomas that contain at least a 25% cystic component and correlated the images with the final diagnosis. Features of the solid component assessed included its echo texture, overall appearance, shape, size, and internal homogeneity. RESULTS: One-hundred seventy-seven teratomas were benign, and 11 were malignant; among the malignant masses, 7 were high grade. Of the benign forms, 155 solid components (88%) were hyperechoic, 168 (95%) were focal in appearance, 105 (59%) were nodular in shape, and 123 (69%) were uniformly solid. Of the malignant types, 9 solid components (82%) were isoechoic, 6 (55%) had branching, 6 (55%) were irregular in shape, and 8 (73%) were uniformly solid. Five malignant teratomas (45% overall and 71% of high-grade subtypes) had branching isoechoic components. Only 2 benign teratomas (1%) had isoechoic components that branched. CONCLUSIONS: The presence of a branching isoechoic component in a cystic ovarian teratoma may suggest malignancy. PMID- 11503923 TI - Sonographic diagnosis of pelvic adhesions in patients after ovum pickup. AB - OBJECTIVE: To evaluate the use of transvaginal sonography for the detection of pelvic adhesions by using clear free fluid in the pouch of Douglas found after ovum pickup. METHODS: A prospective clinical study was performed in an infertility unit of an academic research facility. Sonography was performed in 50 women with infertility 3 days after ovum pickup, and the visceral peritoneum of the uterus, the ovaries, and the fallopian tubes was scanned for possible pelvic adhesions. RESULTS: The serosal surfaces of the uterus, ovaries, and fallopian tubes were successfully observed for the presence of adhesions in 86%, 68%, and 20% of the patients, respectively. Improved visualization was associated with an increased amount of pelvic fluid for the uterus (P = .01) but not for the ovaries and fallopian tubes. The amount of fluid in the pelvis correlated with an increased number of retrieved oocytes (P = .07) and a decreased need for manual manipulation to achieve proper imaging of the uterus (P = .001). CONCLUSION: Transvaginal sonography performed in the presence of fluid in the pelvis may show adhesions mainly attached to the uterus and ovaries. Assessment of possible mechanical factors is important in planning treatment of patients with infertility. PMID- 11503924 TI - Sonohysterography for the diagnosis of residual trophoblastic tissue. AB - OBJECTIVE: To assess the efficacy, safety, and associated complications of sonohysterography for the diagnosis of residual trophoblastic tissue. METHODS: We conducted a prospective study of 23 consecutive patients admitted to our ultrasonography unit with clinical and ultrasonographic signs of retained intrauterine tissue. RESULTS: Twelve patients had hydrosonographic features suggestive of residual trophoblastic tissue (i.e., an intrauterine lesion not detachable from the uterine wall after instillation of saline), whereas in 11 cases the hydrosonographic findings were negative for retained tissue. Blood flow was detected within abnormal intrauterine masses in 4 of 12 patients with trophoblastic tissue, whereas it was not detected in any patient without retained tissue (P = .093). No complications were encountered during the procedure or the postprocedure period. None of the patients had anesthetic complications, perforation of the uterus, fluid overload, or any other surgical complication. All 12 patients underwent hysteroscopic removal of the suspected residual trophoblastic tissue, and histologic confirmation of residual trophoblastic tissue was obtained in all cases. CONCLUSIONS: Sonohysterography for detection and diagnosis of residual trophoblastic tissue is an accurate and safe procedure. Further studies comparing the efficacy of sonohysterography with that of diagnostic hysteroscopy are warranted. PMID- 11503925 TI - Fetal stimulation by pulsed diagnostic ultrasound. AB - OBJECTIVE: To show that pulsed ultrasound from a clinical ultrasonic imaging system can stimulate the fetus. Stimulation is defined mainly as increased fetal gross body movements in response to excitation. METHODS: Fetuses of a group of 9 volunteer women (mean gestational age, 33.37 weeks; range, 25-40 weeks) were evaluated for body movement under 3 different conditions: (1) control, with no ultrasound exposure; (2) ultrasound in continuous wave Doppler mode; and (3) pulsed ultrasound in pulsed Doppler and B modes. A conventional external fetal monitor, with negligible ultrasonic output, was used to monitor fetal gross body motions. After an initial rest period of 3 minutes with 1 or no fetal motion, fetuses were monitored for an additional 3 minutes under the exposure criterion defined for each condition. Resulting fetal motions under the 3 conditions were compared using the Wilcoxon signed rank test. RESULTS: The test showed that fetuses moved significantly more frequently under condition 3 (mean +/- SD, 3.43 +/- 1.93 movements per minute) than under condition 1 (0.40 +/- 7.33 movements per minute) or condition 2 (0.63 +/- 7.67 movements per minute); P = .004 and .016, respectively. Fetal movements under conditions 1 and 2 did not differ significantly. CONCLUSION: Diagnostic ultrasound may stimulate fetal body motion. PMID- 11503926 TI - An approach to acoustic properties of biological tissues using acoustic micrographs of attenuation constant and sound speed. AB - OBJECTIVE: To develop a method for two-dimensional measurement of acoustic properties of biological tissue elements at the microscopic level for specimens with a thickness of approximately 10 microm. METHODS: The procedure was developed on the basis of mechanically scanned acoustic microscopic techniques in the frequency range of 100 to 200 MHz and the theory of interference phenomena. Various tissues and samples were prepared to evaluate the method and to interpret sound propagation properties. RESULTS: Tissues with high protein content, low water content, or both had a high attenuation constant and sound speed. The exponent n of attenuation against frequency was almost unity at the microscopic level, whereas it was greater than unity when the specimen thickness was greater. Sound speed dispersion was not observed. CONCLUSIONS: The method was shown to be reproducible, and the data were interpreted acoustically and pathologically with reference to tissue type and specimen thickness. PMID- 11503927 TI - Translabial sonography of vaginal fibroids: report of 2 cases and review of the literature. AB - OBJECTIVE: To determine the role of translabial sonography in the diagnosis of vaginal fibroids. METHODS: Two women with vaginal masses of undetermined origin were examined by various imaging procedures, including translabial sonography. RESULTS: Initial examinations, which included transabdominal sonography, cystoscopy, and cystourethrography, yielded inconclusive findings. Translabial sonography, however, suggested isolated vaginal leiomyomas in both patients, and in both the diagnosis was confirmed histologically after surgery. CONCLUSIONS: Translabial sonography should be considered as an adjunct to transabdominal and transvaginal sonography for patients with suspected vaginal fibroids. PMID- 11503928 TI - Sonographic features of ovarian cystadenofibromas: spectrum of findings. AB - OBJECTIVE: To describe the sonographic characteristics of ovarian cystadenofibromas. METHODS: We conducted a retrospective study of 23 ovarian tumors histopathologically confirmed as ovarian cystadenofibromas that were preoperatively evaluated by transvaginal color Doppler sonography. RESULTS: In all cases the mass was predominantly cystic. Septations appeared in 30.4% of the tumors. Papillary projections or solid nodules appeared in 56.5% of the cases. The most frequent appearance was a unilocular complex cystic mass. In 47.8% of the tumors, vascularization was detected, having a typical pattern of peripheral vascularization with scattered vessels of high blood flow impedance. CONCLUSIONS: We have described the sonographic spectrum of findings of ovarian cystadenofibromas. The most frequent appearance was a unilocular cystic mass with gross papillary projections or solid nodules. In almost half of the tumors, vascularization could be detected. PMID- 11503929 TI - Sonographic diagnosis of ulnar artery aneurysm in hypothenar hammer syndrome: report of 2 cases. PMID- 11503930 TI - A false-positive diagnosis of a prenatal encephalocele on transvaginal ultrasonography. PMID- 11503931 TI - Lymphoma of the penis: sonographic findings. PMID- 11503932 TI - Synchronous bilateral testicular torsion in an adult. PMID- 11503933 TI - Do ordinary spectacle lenses reduce eye injuries? PMID- 11503934 TI - Overnight orthokeratology. PMID- 11503935 TI - New equations for spectacle induced ocular deviations: responses to some typical questions. PMID- 11503936 TI - Amelanotic choroidal nevus and melanoma: cytology, tumor size, and pigmentation as prognostic indicators. AB - BACKGROUND: Choroidal nevi are fairly common lesions of the posterior pole that can sometimes transform into melanoma, and it is thought that most choroidal melanomas arise from preexisting nevi. Occasionally, these lesions present as nonpigmented or amelanotic variations of their pigmented counterparts. Recent studies suggest a relationship between tumor pigmentation and risk of growth and metastasis, with a better prognosis for lightly pigmented or amelanotic lesions. CASE REPORTS: A case of an amelanotic choroidal nevus and melanoma are presented. In Case 1, a 26-year-old white female was found to have a large amelanotic nevus in the right eye. After 7 years of periodic observation, the lesion has not changed. In Case 2, a 51-year-old white male was diagnosed with a large amelanotic melanoma in the left eye. Due to extensive involvement of the optic nerve, the patient underwent enucleation. Histological evaluation confirmed the lesion as a mixed-cell type malignant amelanotic melanoma. CONCLUSION: Management of choroidal nevi generally consists of periodic observation, and the most widely accepted management of choroidal melanoma is observation, radiotherapy, and transpupillary thermotherapy or enucleation. The therapeutic modality of choice for melanoma will vary depending on the size, growth, and location of the lesion. In addition, recent studies suggest an association between heavy tumor pigmentation, tumor size, cell type, and risk of metastasis. Although many variables will influence the final treatment option, pigmentation of the lesion should also be considered. PMID- 11503937 TI - Acquired unilateral visual loss attributed to an accommodative spasm. AB - BACKGROUND: Accommodative spasm (AS) has been reported to be a rare cause of an acquired unilateral visual loss. We describe a unique case of acquired visual loss due to an apparent unilateral AS triggered by occlusion of the contralateral eye. Clinical examination and photorefraction techniques were performed during the acute phase of the disorder and in the follow-up evaluations. CASE REPORT: An otherwise healthy 27-year-old woman presented with a 2-month decrease of vision in her left eye. Unaided visual acuity in the right eye was 20/20 and in the left eye was finger counting. Orthophoria existed at distance and near. The decrease in vision in the left eye was attributed to an apparent unilateral AS of 5 D, which occurred only when the right eye was occluded. Under the occluder, the right eye also manifested an AS. With the left eye occluded or with binocular viewing, the right eye and left eye accommodated normally. With a 5 D convex lens before the right eye, the visual acuity in the left eye was 20/20. Refractive error with cycloplegia revealed low hyperopia in each eye. CONCLUSION: AS can be the cause of acquired unilateral visual loss. To our knowledge, this is the first documented report of an apparent unilateral AS triggered by occlusion of the contralateral eye. PMID- 11503938 TI - Prospective quantification of near work using the experience sampling method. AB - INTRODUCTION: The assessment of individuals' near work by survey methods is challenging. The feasibility of the Experience Sampling Method to quantify daily visual tasks was evaluated. METHODS: Twenty-one subjects were randomly paged five times per day for 6 days. When paged, the subjects dialed into a telephone survey to report the nature, duration, and working distance of their visual activity at the time of the page. RESULTS: The overall response rate was 81.3% (512/630 pages). The individual response rates varied from 6.7% to 100% (median, 86.7%). Responses were grouped into 11 categories/activities for ease of analysis. Of 506 responses, the most common activity category was "distance tasks," which included driving and walking (N = 92). Other common responses included computer use (N = 68), reading (N = 66), household tasks, e.g., cleaning, cooking, and showering (N = 64), and watching television (N = 61). Activities with a mean distance < or =26 inches (arm length) were combined as near-work responses and accounted for 54.3% (258/475) of all responses. CONCLUSION: A modification of the Experience Sampling Method can be used to obtain a "real-time" sampling of visual activities. PMID- 11503939 TI - Effect of induced anisometropia on binocular through-focus contrast sensitivity. AB - PURPOSE: The aim of this study was to characterize binocular through-focus function in white light and to investigate the effect of induced anisometropia on binocular depth of focus. METHODS: The subjects viewed sine-wave gratings generated on a monitor through a modified Badal system that produced gradual changes in target vergence ranging from -4.00 to +2.00 D. Binocular through-focus contrast sensitivity curves were obtained at a spatial frequency of 11 cpd and for different levels of induced anisometropia. Subjective depths of focus were derived from the through-focus curves. RESULTS: An induced anisometropia lower than 1.00 D led to a monomodal through-focus curve involving a single depth of focus, whereas with higher anisometropia, the curves became bimodal indicating a lack of performance at intermediate distance. Binocular thresholds predicted by the quadratic summation model from our monocular measurements were well correlated to our binocular measurements. Predictions allowed us to estimate optimum levels of induced anisometropia. PMID- 11503940 TI - All developmental dyslexic subtypes display an elevated motion coherence threshold. AB - PURPOSE: Psychophysical studies indicate that many dyslexics have a motion processing deficit. The purpose of this study was to determine whether elevated motion coherence thresholds correlate with the specific dyslexic subtypes as defined by the Boder classification scheme. METHODS: Twenty-one dyslexics (seven dyseidetics, six dysphonetics, and eight dysphoneidetics) and 19 age- and gender matched controls participated in the study. The dyslexics were identified by an exclusionary approach and then subtyped with the Adult Dyslexia Test or the Dyslexia Determination Test. Motion coherence thresholds were determined with random dot kinematograms composed of signal dots and noise dots. Signal dots moved either left or right on each trial, whereas noise dots moved in random directions. The percentage of dots that comprised the signal was varied randomly on each trial (0 to 21% in 3% increments). Subjects guessed the direction of signal dot motion on each trial (two-alternative forced-choice task). A 75% correct threshold was determined with a Weibull equation fit to the psychometric function. RESULTS: All three dyslexic subtypes had elevated motion coherence thresholds (t-test; dyseidetics p = 0.01, dysphonetics p = 0.039, dysphoneidetics p = 0.048). CONCLUSION: Motion-coherence deficits are not correlated with a specific dyslexic subtype, but, rather, are common to all subtypes. However, some individuals in each of the dyslexic subtypes were found to have normal motion coherence thresholds, suggesting that other factors must be considered to predict the motion sensitivity deficits found in dyslexia. PMID- 11503941 TI - Resistance of plastic ophthalmic lenses: the effect of base curve on different materials during static load testing. AB - PURPOSE: This study was designated to evaluate, through a static load test, the influence of lens base curve on the fracture resistance of three common plastic materials. METHODS: A JJ Lloyd load cell machine was used to test the fracture resistance of -4.00 D spherical lenses. The samples had a nominal center thickness of 2.0 mm and a base curve distributed in one of five groups (+0.50, +2.50, +4.50, +6.50, and +8.50 D). The lenses were manufactured in CR39, polycarbonate, and TL16, a high refractive index plastic (n = 1.599). RESULTS: The lens base curve influenced fracture resistance for all materials. For these materials, resistance increased as the base curve varied from +0.50 to +8.50 D. The resistance of CR39, TL16, and polycarbonate lenses was found to be linearly dependent on lens base curve. The effect is stronger for polycarbonate. Fracture resistance was higher for TL16 than for CR39, and polycarbonate was much more resistant to breakage than the two other materials. CONCLUSIONS: For a given power, the fracture resistance of an ophthalmic lens is reduced when its base curve has a low value. Consequently, the flattening of ophthalmic lenses for cosmetic purposes is not recommended as far as fracture resistance is concerned. PMID- 11503942 TI - The power output of laser pointers: do they exceed federal standards? AB - BACKGROUND: Concern about the safety of laser pointers has increased as these devices have become readily available to the public, which includes children who may use them improperly. According to federal (Food and Drug Administration) standards, laser pointers are class 3a devices, and power output cannot exceed 5.0 mW. METHODS: We tested a sample consisting of 29 red diode laser pointers obtained from various sources that were equipped with fresh batteries. Power output was measured with an optical power meter with the sensor 5.0 cm from the laser. The laser was turned on for 60 s and a reading was taken every 10 s. This procedure was repeated five times for every laser. RESULTS: The power output of unaltered lasers ranged from 0.7 mW to a high of 3.9 mW. Alterations to some of the lasers increased the output slightly above 5.0 mW. CONCLUSIONS: When the laser pointers in our sample were used as packaged, they did not exceed the federal standards of 5.0 mW. PMID- 11503943 TI - Modeling logMAR visual acuity scores: effects of termination rules and alternative forced-choice options. AB - BACKGROUND: Logarithm of the minimum angle of resolution (logMAR) charts come in a variety of alternative forced-choice (AFC) formats and can be used with a variety of different rules to determine when to stop a subject reading down a chart (termination rules). METHODS: Exact calculation and Monte Carlo simulation techniques were used to compare logMAR scores for different termination rules and for infinite-, 26-, 10-, 8-, 4-, and 2-AFC logMAR chart formats. Slope-corrected standard deviation, an index of variability of the underlying ideal threshold, was used as a common metric for the different test conditions. RESULTS: The mean and standard deviation of logMAR scores can be significantly affected by termination rules and AFC format. For different AFC formats, different termination criteria were found to give optimal slope-corrected standard deviations. CONCLUSION: A number of clinically suitable termination rules are proposed for different AFC format logMAR charts. For letter-by-letter scoring of Bailey-Lovie and EDTRS charts, it is recommended that researchers and practitioners use a termination rule of stopping after four or more mistakes on a line. PMID- 11503944 TI - Compensating for vertical anisometropic imbalance by the positioning of segment centers. AB - Prismatic imbalance produced in the reading area by an anisometropic spectacle correction can be offset by various methods. If a reading addition is present, one method is to provide differently shaped segments and allowing their optical centers to be separated vertically while the segment tops remain in horizontal alignment. Traditionally, to apply this method, the relative prismatic difference at the reading point is first determined by a thin lens application of Prentice's rule. This rule is then applied a second time to determine the placement of the segments that will offset the prismatic difference produced by the major lenses. However, the conventional application of Prentice's rule for determining the fusional demand in a particular area of the spectacle field often produces very large errors, which will affect the calculation of the placement of the segments. This article develops and demonstrates more accurate methods for applying the principle of compensating segments. PMID- 11503945 TI - Enhancement of heart rate variability by cholinergic stimulation with pyridostigmine in healthy subjects. AB - The purpose of this study was to determine the effect of the oral administration of pyridostigmine bromide on indices of heart rate variability (HRV) in healthy young volunteers. Seventeen healthy participants (11 men, 6 women; aged 27 +/- 8 y) submitted to a randomized, crossover, double-blind protocol, in which they received 30 mg pyridostigmine bromide (PYR) or placebo orally at 8-hour intervals for 24 hours, on two separate days. Venous blood samples were collected 2 and 24 hours after the first dose for determination of serum cholinesterase activity. Holter tapes were recorded during the 24-hour period and analyzed using a semiautomatic technique to evaluate time- and frequency-domain indices of HRV and to build three-dimensional return maps for later quantification. Symptoms were mild and occurred similarly during administration of PYR and placebo (p = 0.140). Serum cholinesterase activity was reduced by 15% at 2 hours (p = 0.013) and by 14% at 24 hours (p = 0.010) after the first dose of PYR, but not after administration of placebo. Pyridostigmine administration caused a significant increase in the mean 24-hour R-R interval (placebo: 814 +/- 20 msec; PYR: 844 +/- 18 msec; p = 0.003) and in time-domain indices of HRV, such as the standard deviation of all R-R intervals (SDNN; placebo: 151 +/- 9 msec; PYR: 164 +/- 9 msec; p = 0.017), and the percentage of pairs of adjacent R-R intervals differing by more than 50 msec (pNN50; placebo: 12.8 +/- 1.8%; PYR: 13.9 +/- 1.5%; p = 0.029). Pyridostigmine had no significant effect on frequency-domain indices of HRV, but resulted in significant increase in P2, a parasympathetic index derived from the three-dimensional return map (placebo: 93 +/- 13 msec; PYR: 98 +/- 13 ms; p = 0.029). In conclusion, low-dose pyridostigmine reduced mean heart rate and increased HRV during a 24-hour period in healthy young subjects. PMID- 11503946 TI - Spectral and cross-spectral autoregressive analysis of cardiovascular variables in subjects with different degrees of orthostatic tolerance. AB - The mechanisms leading to vasovagal syncope are still unclear. A simple discriminating test for the identification of syncope-prone subjects is not presently available. Fifty-two subjects had a stepwise orthostatic test with 60 degrees tilt and -20 and -40 mm Hg lower-body negative pressure before the appearance of impending syncope symptoms. Spectral and cross-spectral analyses of heart period and systolic pressure time series were performed to estimate the power of the high-frequency (approximately equals 0.25 Hz) and low-frequency (approximately equals 0.1 Hz) oscillations, the coherence between heart period and systolic pressure, and the mean low-frequency and high-frequency central frequency, phase shift, and transfer function at maximal coherence. According to time to presyncope, the 52 subjects were divided into two groups: 25 with normal orthostatic tolerance, and 27 with poor orthostatic tolerance. In the supine positions, the mean central low-frequency was significantly lower in poor tolerance group than in normal-tolerance group, discriminating poor from normal orthostatic tolerance with 80% specificity and 83% sensitivity, and was significantly correlated to time to presyncope. In the 2 to 3 minutes preceding syncope, subjects with poor orthostatic tolerance had less tachycardia, lower low frequency power of systolic pressure, higher respiratory frequency, and a less negative phase shift in high-frequency range. In presyncope, sympathetic activation is reduced in subjects with poor orthostatic tolerance. In addition, the higher breathing frequency and the smaller negativity of phase shift in high frequency range, which may indicate an inadequate engagement of the baroreflex, suggest a causal role of respiration in the development of syncope. Supine central values of low frequency may be proposed as a valuable clinical index of orthostatic intolerance. PMID- 11503947 TI - The effects of autonomic dysfunction and endurance training on cardiovascular control. AB - The effects of autonomic dysfunction and regular activity on the cardiovascular system were investigated. The 48 participants included 12 subjects with tetraplegia, 12 subjects with paraplegia, 12 sedentary subjects, and 12 endurance trained able-bodied controls. Central and peripheral autonomic data were obtained at rest to estimate efferent cardiac vagal output and sympathetic vasomotor control, and plasma norepinephrine concentration was determined as a marker of peripheral sympathetic activity. Cardiovascular parameters were obtained using a noninvasive cardiac output maneuver. The group with paraplegia did not differ from the sedentary group for efferent cardiac vagal output, but all other group comparisons were different (p <0.05). Sympathetic vasomotor control and stroke index were also similar between the paraplegia and sedentary groups, whereas both were increased in the endurance-trained group and were significantly reduced in the tetraplegia group. A strong relation between efferent cardiac vagal output and stroke index was established for the total group (r = 0.78, p <0.01), and analysis of covariance determined that the slope of this relation was similar among the groups. Sympathetic vasomotor control correlated significantly with plasma norepinephrine (r = 0.57, p <0.01), and a relation between sympathetic vasomotor control and stroke index was identified for the total group (r = 0.40, p <0.01). These results suggest that vagal control of resting central cardiac function is maintained despite autonomic dysfunction. The comparable findings in the paraplegia and sedentary groups suggest that regardless of peripheral autonomic dysfunction, the absence of regular physical activity has a similar effect on the resting vagal modulation and stroke index. PMID- 11503948 TI - Brain meets gut: gastroesophageal reflux. PMID- 11503949 TI - Cerebrovascular and cardiovascular responses associated with orthostatic intolerance and tachycardia. AB - Idiopathic orthostatic intolerance syndrome is characterized by postural symptoms of cerebral hypoperfusion without arterial hypotension. Abnormal baroreceptor responses with deranged cerebral autoregulation leading to cerebral vasoconstriction have been proposed as a causative mechanism. The authors report the cerebrovascular and cardiovascular responses in a patient who recovered from orthostatic intolerance and tachycardia. Changes in the orthostatic responses of mean arterial pressure (MAP), heart rate (HR), cardiac output (CO), and transcranial Doppler middle cerebral artery (MCA) mean blood flow velocity (Vmean) were assessed at admission and again 6 months after recovery. Normal cardiovascular responses to forced breathing and to standing indicated intact overall baroreflex integrity with normal baroreflex sensitivity (10.2 msec.mm Hg( 1)). After the patient stood for 8 minutes, presyncopal symptoms developed, with unchanged MAP but increased HR (+41 beats/min) and reduced stroke volume (SV) ( 69%), CO (-50%), and MCA Vmean (-46%; 57 to 31 cm. s(-1)). After a reconditioning program and recovery, the patient was reexamined. The supine MCA Vmean was larger (79 cm. s(-1)), as were MAP (76 versus 70 mm Hg) and CO (+15%). The orthostatic HR increase was smaller (+5 beats/min), as was the reduction in SV (-44%) and CO (-30%), with an increase in MAP to 93 mm Hg. The orthostatic reduction in MCA Vmean was smaller (-13 versus -26 cm.s(-1)) and standing cerebrovascular resistance decreased (1.41 versus 2.39 mm Hg.cm. s(-1)). In this patient who had intact baroreflex control and no postural decrease in blood pressure, the reduction in MCA Vmean, concomitant with a large decrease in CO, seemed reversible. The result suggests that a symptomatic reduction in cerebrovascular conductance during standing is to be interpreted as being an adaptive response to a critically limited systemic blood flow, rather than to derangement of cerebral autoregulation. PMID- 11503950 TI - Assessment of cardiovascular autonomic dysfunction in multiple system atrophy. AB - Because heart rate is controlled mainly by the autonomic nervous system, cardiovascular autonomic dysfunction may contribute to the prognosis of patients with multiple system atrophy (MSA). To clarify cardiovascular autonomic dysfunctions in MSA, the authors investigated the relation between blood pressure (BP) and pulse rate (PR), and assessed a power spectral analysis of heart rate variability (HRV) during the clinical course using ambulatory BP and a heart rate monitor for 24 hours. The authors studied seven patients with MSA (five men and two women, aged 61.0 +/- 5.8 years) and seven healthy volunteers (four men and three women, aged 58.0 +/- 6.6 years) without hypertension, heart disease, or intracranial lesions. The MSA group showed abnormal circadian variations of BP and PR and a significantly decreased correlation coefficient between BP and PR. A significant decrease and altered circadian variation also existed in the number of changes in successive R-R intervals greater than 50 msec (RR50) and in the power of the high- and low-frequency component of HRV. The authors observed a significant negative correlation between the duration of illness and the number of changes in successive R-R intervals greater than 50 msec. The characteristic dysautonomia in MSA was a decrease in sympathetic and parasympathetic activity, with an abnormal circadian rhythm of BP and HRV. The balance between sympathetic and parasympathetic activity was also impaired. The parasympathetic modulation represented by RR50 worsened according to the development of the illness. Those autonomic dysfunctions may have affected the cardiovascular systems, which may indicate a poor prognosis in patients with MSA. An analysis of HRV and the circadian rhythm of BP and HRV are useful in evaluating cardiac autonomic dysfunctions in MSA. PMID- 11503951 TI - Decreased sympathetic inhibition in gastroesophageal reflux disease. AB - This study was undertaken to evaluate autonomic nervous system function in patients with gastroesophageal reflux disease. Based on clinical criteria, 28 consecutive patients with no history of heart, metabolic, or neurologic disease (mean age 41 y, range 20-62 y) reporting with upper gastrointestinal symptoms typical of gastroesophageal reflux underwent esophageal manometry, ambulatory 24 hour pH study with electrocardiographic monitoring, power spectral analysis of heart rate variability, and cardiovascular tests. Twelve healthy subjects served as controls. A positive result of prolonged esophageal pH study (pH in the distal esophagus less than 4, lasting more than 4.2% of recording time) was observed in 21 patients (reflux group); seven patients were categorized in the nonreflux group. No patient showed arrhythmias or any correlation between heart rate variability changes during electrocardiographic monitoring and episodes of reflux (pH less than 4, lasting more than 5 minutes). A decrease of sympathetic function occurred only in the reflux group (p <0.05) supported by the lower increase of systolic/diastolic blood pressure at sustained handgrip. No other cardiovascular tests showed statistically significant differences in the control or nonreflux groups. Total time reflux showed an inverse correlation with sympathetic function in the reflux group (r = -0.415, p <0.028). We concluded that there is some evidence for a slightly decreased sympathetic function in patients with gastroesophageal reflux disease that is inversely correlated with total time reflux. In these patients, decreased sympathetic function may cause dysfunction of intrinsic inhibitory control with increased transient spontaneous lower esophageal sphincter relaxations, thus resulting in gastroesophageal reflux disease. PMID- 11503952 TI - Sleep embodies maximum and minimum levels of autonomic integration. AB - This review is based on the Valsalva lecture delivered by Professor Elio Lugaresi at the first meeting on the European Federation of Autonomic Societies, held in conjunction with the Italian Autonomic Society in Bologna in July 1999. PMID- 11503953 TI - Epidemiological features of myelodysplastic syndromes: results from regional cancer surveys and hospital-based statistics. AB - Although myelodysplastic syndromes (MDS) have been increasingly diagnosed in recent years, precise data on their prevalence and incidence are still lacking. Due to difficulties of diagnosis and classification, large-scale population-based studies that are required for obtaining truly representative data on the epidemiology of MDS are currently not available. Our present knowledge of the incidence and other epidemiological characteristics of MDS is based on a few regional studies performed by authors with a long-lasting interest in these hemopathies. Despite certain limitations, these studies have consistently shown that MDS are relatively common hematological malignancies. Their crude incidence varies from 2.1 to 12.6 cases per 100,000 people per year. Among the age group that is mainly affected, people older than 70 years, we are now faced with incidence rates of about 15 to 50 cases per 100,000 people per year. The recent increase in MDS incidence observed in some studies is probably not the result of an actual increase in the number of cases, but reflects improvements in geriatric medical care and diagnosis. PMID- 11503954 TI - Clonality in the myelodysplastic syndromes. AB - The myelodysplastic syndromes (MDSs) comprise a heterogeneous group of stem cell disorders involving cytopenia and dysplastic changes in 3 hematopoietic lineages. Although it is accepted that MDS is a clonal disorder, the exact nature of the involvement of multipotent stem cells and progenitor cells has not been resolved. Most clonality studies of MDS support the proposal that the primary neoplastic event occurs, in most patients, at the level of a committed myeloid progenitor cell, capable of differentiation into multiple myeloid lineages. The extent of the involvement of T and B lymphocytes in MDS remains controversial. Much of the variation reported may result from disease heterogeneity and technical issues such as skewed methylation patterns occurring in studies analyzing X-chromosome inactivation patterns (XCIP) and possible impurities in lymphocyte preparation. A great deal of the evidence in support of T-lymphocyte involvement in MDS has been generated by XCIP studies, and some of these data need to be treated with caution, especially data from studies in which appropriate controls were omitted. In contrast, B-lymphocyte involvement in some patients with MDS is based on studies using more robust technology including combined immunophenotyping and fluorescence in situ hybridization. Clonality studies involving myeloid and lymphoid cells in MDS have yielded discrepant results with regard to the potential involvement of multipotent (lympho-myeloid) hematopoietic stem cells (HSCs). However, failure to detect a clonal marker in all cells of all lineages does not preclude multipotent-HSC involvement. Some recent studies have produced compelling evidence to show that, in some patients with MDS, the multipotent HSC is the target of the primary neoplastic event. It now seems probable that MDS arises in multipotent HSCs more commonly than previously recognized. Such data not only provide important new insights into the biology of MDS but also may have therapeutic implications. The determination of whether multipotent HSCs are involved in the MDS clone may be important for the use of autologous stem cell transplantation in these patients. PMID- 11503955 TI - The role of apoptosis in the pathogenesis of the myelodysplastic syndromes. AB - The paradoxical occurrence of peripheral cytopenias despite a normo/hypercellular marrow in myelodysplastic syndromes (MDS) has been attributed to excessive intramedullary hematopoietic cell apoptosis. It has also been postulated that abrogation of programmed cell death (PCD) may underlie MDS transformation to acute myeloid leukemia (AML). Despite overwhelming evidence for a role of aberrant apoptosis in myelodysplasia, the molecular mechanisms responsible for such changes have not been elucidated. This paper summarizes current evidence implicating a role for altered PCD in MDS and outlines potential cellular mechanisms whereby hematopoietic progenitor cell apoptosis may be dysregulated. PMID- 11503956 TI - Chromosome and molecular abnormalities in myelodysplastic syndromes. AB - Cytogenetic abnormalities are seen in approximately 50% of cases of myelodysplastic syndrome (MDS) and 80% of cases of secondary MDS (following chemotherapy or radiotherapy). These abnormalities generally consist of partial or complete chromosome deletion or addition (del5q, -7, +8, -Y, del20q), whereas balanced or unbalanced translocations are rarely found in MDS. Fluorescence hybridization techniques (fluorescence in situ hybridization [FISH], multiplex FISH, and spectral karyotyping) are useful in detecting chromosomal anomalies in cases in which few mitoses are obtained or rearrangements are complex. Ras mutations are the molecular abnormalities most frequently found in MDS, followed by p15 gene hypermethylation, FLT3 duplications, and p53 mutations, but none of these abnormalities are specific for MDS. The rare cases of balanced translocations in MDS have allowed the identification of genes whose rearrangements appear to play a role in the pathogenesis of some cases of MDS. These genes include MDS1-EVI1 in t(3;3) or t(3;21) translocations, TEL in t(5;12), HIP1 in t(5;7), MLF1 in t(3;5), and MEL1 in t(1;3). Genes more frequently implicated in the pathogenesis of MDS cases, such as those involving del5q, remain unknown, although some candidate genes are currently being studied. Cytogenetic and known molecular abnormalities generally carry a poor prognosis in MDS and can be incorporated into prognostic scoring systems such as the International Prognostic Scoring System. PMID- 11503958 TI - Bone marrow colony-forming unit assay in cats with naturally occurring myelodysplastic syndromes. AB - Feline myelodysplastic syndromes (MDS) has been diagnosed in many cats infected with feline leukemia virus, although the pathogenesis of this hematopoietic deficiency has been unclear. In this study, we assayed the bone marrow erythroid colony-forming units (CFU-E) and granulocyte-machrophage CFUs (CFU-GM) to investigate the pathogenesis of feline MDS. The number of CFU-E colonies was decreased in 4 of 7 cats with MDS, and the number of CFU-GM colonies was also decreased in 4 cats. Furthermore, small colonies of CFU-GM were found in all 7 cases. These findings indicated that refractory cytopenia of feline MDS could be caused by abnormal maturation and differentiation of hematopoietic stem cells in bone marrow, as it is in human MDS. The pathogenesis of feline MDS might be similar to that of human MDS. PMID- 11503959 TI - Hematopoietic capability of CD34+ cord blood cells: a comparison with CD34+ adult bone marrow cells. AB - The characteristics of hematopoietic progenitor and stem cell (HPC/HSC) populations in mammals vary according to their ontogenic stage. In humans, HPC/HSCs from umbilical cord blood (CB) are increasingly used as an alternative to HPC/HSCs from adult bone marrow (BM) for the treatment of various hematologic disorders. How the hematopoietic activity of progenitor and stem cells in CB differs from that in adult BM remains unclear, however. We compared CD34+ cells, a hematopoietic cell population, in CB with those in adult BM using phenotypic subpopulations analyzed by flow cytometry, the colony-forming activity in methylcellulose clonal cultures, and the repopulating ability of these cells in NOD/Shi-scid (NOD/SCID) mice. Although the proportion of CD34+ cells was higher in adult BM than in CB mononuclear cells, the more immature subpopulations, CD34+ CD33- and CD34+ CD38- cells, were present in higher proportions in CD34+ CB cells. Clonal culture assay showed that more multipotential progenitors were present in CD34+ CB cells. When transplanted into NOD/SCID mice. CD34+ adult BM cells could not reconstitute human hematopoiesis in recipient BM, but CD34+ CB cells achieved a high level of engraftment, indicating that CD34+ CB cells possess a greater repopulating ability. These results demonstrated that human hematopoiesis changes with development from fetus to adult. Furthermore, CD34+ CB cells contained a greater number of primitive hematopoietic cells, including HSCs, than did adult BM, suggesting the usefulness of CD34+ CB cells not only as a graft for therapeutic HSC transplantation but also as a target cell population for ex vivo expansion of transplantable HSCs and for gene transfer in gene therapy. PMID- 11503957 TI - Differentiation, apoptosis, and function of human immature and mature myeloid cells: intracellular signaling mechanism. AB - Human myeloid cells include hematopoietic cells at various stages of differentiation, from immature myeloid cells to mature phagocytes. Normal immature myeloid cells undergo differentiation concomitantly with proliferation in response to hematopoietic growth factors, and terminally differentiated cells, ie, mature phagocytes, exert their effector functions and then die a natural death via apoptosis. However, leukemic myeloid cells are induced to differentiate with growth suppression by several inducers, such as retinoic acid. This review describes differentiation, apoptosis, and functionality of human myeloid cells. mainly focusing on the intracellular signaling mechanism. The signal transduction system for these biological events of the life cycle of myeloid cells has recently been studied, and several characteristics have been elucidated. First, the signaling pathway for myeloid differentiation is mainly focused in the mitogen-activated protein kinases, such as extracellular signal-regulated kinase and p38, and transcriptional factors such as the signal transducers and activators of transcription PU.1 and CCAAT enhancer binding protein. Second, the signaling mechanism for myeloid cell apoptosis is fundamentally identical to that found in other cells. Caspases, caspase-activated DNase, and mitochondrial molecules such as apoptosis-inducing factor have been reported to be important, and mitogen-activated protein kinases such as p38 appear to be less important. Finally, p38 and phosphatidylinositol 3-kinase play critical roles in the signaling cascade for functional activation of mature phagocytes. The reasons why the same signaling molecules play distinct roles according to the differentiation stage and biological event await future clarification. PMID- 11503960 TI - Defective megakaryopoiesis and abnormal erythroid development in Fli-1 gene targeted mice. AB - Mouse embryos homozygous for a targeted disruption in the Fli-1 gene show hemorrhage into the neural tube and brain on embryonic day (E)11.0 and die shortly thereafter. Livers from the mutant embryos contain drastically reduced numbers of pronormoblasts, basophilic normoblasts, and colony-forming cells. To determine the nature of impaired hematopoiesis, we carried out cell culture studies of mutant embryonic stem (ES) cells and cells from the aorta-gonad mesonephros (AGM) region of E10.0 mutant embryos. There was a striking reduction in the number of megakaryocytes in cultures of mutant AGM cells compared with cultures of AGM cells from wild-type or heterozygous embryos. Furthermore, Fli-1 mutant ES cells failed to produce megakaryocyte colonies and multilineage colonies containing megakaryocytes. Consistent with the observed defect in megakaryopoiesis, we also demonstrated the down-regulation of c-mpl in the AGM of mutant embryos. The percentages of pronormoblasts and basophilic normoblasts were significantly reduced in cultures of mutant AGM embryos, which contained primarily polychromatophilic and orthochromatic normoblasts. These results provide further evidence for the role of Fli-1 in the regulation of hematopoiesis and for c-mpl as a Fli-1 target gene. PMID- 11503961 TI - Targeted integration of foreign DNA into a defined locus on chromosome 19 in K562 cells using AAV-derived components. AB - Targeted integration of foreign DNA is ideal for gene therapy, particularly when target cells such as hematopoietic cells actively divide and proliferate. Adeno associated virus (AAV) has been shown to integrate its genome into a defined locus, AAVS1 (19q13.3-qter). The inverted terminal repeat (ITR) and Rep proteins are responsible for this site-specific integration, and a system has been developed that delivers a gene preferentially into AAVS1 by using these components of AAV. We examined whether this system could be applied to gene transfer into K562 cells. Two rep expression plasmids were tested, 1 driven by the cytomegalovirus (CMV) promoter (pCMVR78) and the other under the translational control of an internal ribosome entry site (pMGiR78) with mouse mammary tumor virus promoter. K562 cells were cotransfected with a rep plasmid and a plasmid containing a neo gene flanked by the ITRs. G418-resistant clones were isolated and analyzed by Southern blot analysis and fluorescence in situ hybridization (FISH). Southern blot analysis suggested AAVS1-specific integration of the neo gene in 6 (35%) of 17 clones when K562 cells were transfected with pMGiR78 by lipofection. FISH located the neo gene on chromosome 19 in 5 of these 6 clones (29%). Eight (32%) of 25 clones obtained by electroporation with pCMVR78 had the neo gene at AAVS1, according to Southern blot analysis, and 4 of these 8 clones (16%) were positive according to FISH analysis. These results suggest that site-specific integration of foreign DNA can be achieved at a significantly high rate in human hematopoietic cells using the AAV components. PMID- 11503962 TI - Selective transduction of HIV-1-infected cells by the combination of HIV and MMLV vectors. AB - Human immunodeficiency virus 1 (HIV-1)-infected cells are important targets of gene therapy for acquired immune deficiency syndrome. We have developed a novel strategy for targeted gene transfer into HIV-1-infected cells based on 2-step gene transfer. The first step involves the stable introduction of the HIV vector containing the ecotropic Moloney murine leukemia virus (MMLV) receptor gene (EcoRec) into human CD4+ T cells as a molecular switch. Because the HIV-long terminal repeat (HIV-LTR) is Tat inducible, it is expected that EcoRec is expressed only after HIV-1 infection. Northern blot analysis and a retrovirus binding assay confirmed that the HIV-LTR of the integrated vector was silent in transduced cells but strongly transactivated in HIV-1 infection. High levels of EcoRec expression were observed only in HIV-1-infected cells. These cells became highly susceptible to ecotropic MMLV infection and, therefore, in the second step, HIV-1-infected cells were selectively transduced with ecotropic MMLV vectors. More than 70% of HIV-1-infected cells were transduced by this strategy. These findings indicate that this 2-step method can be used for selective and stable gene transfer into HIV-1-infected cells. PMID- 11503964 TI - Paratracheal extramedullary hematopoiesis. AB - Extramedullary hematopoiesis (EMH) is a rare finding in hematology. A 73-year-old female patient with a 1-week history of severe progressive dyspnea was examined, and computed tomography (CT) showed a paratracheal mass 3 cm in size located 1 cm below the vocals cords and causing obliteration of the tracheal airway. Cytology of a needle biopsy revealed EMH. External radiotherapy of 200-cGy fractions to a total dose of 2000 cGy was administered with 3-dimensional conformal planning to treat the progressive symptoms. The patient's clinical symptoms started to improve 2 days after radiotherapy and had completely disappeared after 7 days. CT scans showed complete response on follow-up at 1 week to 5 months after radiotherapy. Mature and immature hematopoietic cells and many adipose cells were seen in the pretreatment samples. Histologic findings in the posttreatment samples showed that these cells had completely disappeared due to the conformal radiotherapy. On the basis of clinical, radiologic, and histologic results, we suggest that conformal radiotherapy may be useful for the treatment of paratracheal localization of EMH because good tumoral irradiation was obtained in this case, with the protection of normal tissues. PMID- 11503963 TI - Reduced CD43 expression on the neutrophils of MDS patients correlates with an activated phenotype of these cells. AB - CD43 (also known as leukosialin and sialophorin) is a surface sialoglycoprotein expressed at high levels on most leukocytes implicated in adhesion, antiadhesion, and activation/proliferation mechanisms. We studied the expression of this molecule on the leukocytes of patients with myelodysplastic syndromes (MDSs) in an effort to detect acquired deficiencies of this molecule. We used immunofluorescence flow cytometry in analyzing whole blood and isolated neutrophils from 49 MDS patients, 33 men and 16 women aged 33 to 85 years (median, 75 years), and 18 healthy individuals aged 35 to 80 years (median, 72 years). According to French-American-British classification criteria, 13 patients had refractory anemia, 18 had refractory anemia with ringed sideroblasts, 9 had refractory anemia with excess of blasts, 4 had refractory anemia with excess of blasts in transformation to acute leukemia, and 5 had chronic myelomonocytic leukemia. We found decreased expression of CD43 on the neutrophils of these patients, and we correlated this finding with the activation status of these cells as it is defined by their phenotypes. We studied the expression of CD11b, CD18, CD35, CD67, CD69, CD44, and CD53 molecules known to be changed in the activated form of neutrophils. CD43 expression correlated positively with CD53 and CD44 expression and negatively with CD11b, CD18, CD35, CD67, and CD69 expression. Additionally, increased levels of soluble vascular cell adhesion molecules were detected in these patients, suggesting endothelial cell activation. In conclusion, we believe that the decreased expression of CD43 on the neutrophils of MDS patients is associated with activation of these cells and is probably due to cleavage of the molecule from the cell surface and that the same mechanism is possibly responsible for the parallel down-regulation of CD44 and CD53. PMID- 11503965 TI - Effect of additional chromosomal abnormalities in acute promyelocytic leukemia treated with all-trans-retinoic acid: a report of 17 patients. AB - Seventeen cases of acute promyelocytic leukemia (APL) treated with all-trans retinoic acid (ATRA) and combination chemotherapy at Tokyo Metropolitan Komagome Hospital between 1992 and 1999 were reviewed, and divided into 2 karyotype-based cytogenetic groups. One group comprised 7 patients with either the typical t(15;17) alone or a normal karyotype, and the other group comprised 10 patients with additional karyotypic abnormalities. No patient had received prior chemotherapy or irradiation, and no cases were complicated by a history of myelodysplastic syndrome before the diagnosis of APL. There were no significant differences in clinical characteristics at disease presentation. Complete remission was achieved in all 17 patients and karyotypes of bone marrow cells normalized in all cases. No differences were found in relapse rate, overall survival, or disease-free survival between the 2 groups. The analysis did not reveal any significant effect of additional chromosomal abnormalities on the prognosis of APL patients undergoing treatment with ATRA. However, a small number of patients were assessed in this study, and further cumulative studies are needed. PMID- 11503966 TI - Significant association between expression of the CD11b surface molecule and favorable outcome for patients with acute myeloblastic leukemia. AB - The expression of several myeloid and non-lineage associated surface antigens in 70 patients with acute myeloblastic leukemia was investigated in relation to patient and disease characteristics, response to therapy, and prognosis. A leukocyte integrin, CD11b, was the only antigen that showed a significant association with complete remission (CR) duration and survival (P < .025). The mean survival for CD11b+ patients was longer than for CD11b- patients (578 +/- 76 versus 397 +/- 7 days, respectively). CR duration was 897 +/- 84 for CD11b+ patients and 366 +/- 71 for CD11b- patients. Multivariate analysis confirmed the predictive value of CD11b expression for longer survival (relative risk, 3.2; P = .02) and CR duration (relative risk, 3.2; P = .03). CR rate was also significantly higher in CD11b+ patients (77.3%) than in CD11b- patients (46.1%) (P = .01). Survival and remission duration were not influenced by expression of other surface markers including CD13, CD14, CD33, CD34, CD71, CD38, and HLA-DR or by other variables including French-American-British subtype, age, and leukocyte count. Extramedullary disease (EXD) was associated with the presence of both CD13 and CD14 expression (P < .04) but occurred less frequently in CD13+ cases. CD13 expression occurred more frequently in female patients (P = .03). CD38 expression was associated with lower platelet count and an increase in the number of blasts (P < .02). PMID- 11503967 TI - Expression of the Gfi-1 gene in HTLV-1-transformed T cells. AB - Human T-cell leukemia virus type I (HTLV-I) is the causative agent of adult T cell leukemia/lymphoma (ATLL) and immortalizes human T cells interleukin-2 (IL-2) dependently in vitro. Protracted culture of HTLV-I-infected T cells enables them to grow IL-2-independently. Although acquisition of IL-2-independent growth has been correlated with activation of signal transducers and activators of transcription (STATs), the precise mechanism of IL-2-independent growth is unknown. We found that expression of the Gfi-1 (growth factor independence-1) gene was elevated in most HTLV-I-transformed IL-2-independent cell lines but in few HTLV-I-infected IL-2-dependent cell lines. We also found elevated expression of Gfi-1 in fresh leukemic cells of ATLL patients. Although expression of Gfi-1 is correlated with activation of STAT3, induction of the dominant negative form of STAT3 in the HUT102 cell line does not alter the level of Gfi-1 expression. Furthermore, MT2 cells treated with Gfi-1 antisense oligonucleotide had reduced [3H]thymidine uptake compared with MT2 cells treated with Gfi-1 sense oligonucleotide. These findings indicate that Gfi-1 activation is involved in the IL-2-independent growth of HTLV-I-transformed T cells in vitro and in the development of ATLL in vivo, but is not induced by STAT activation. PMID- 11503968 TI - Clinical trial to investigate the pharmacokinetics, pharmacodynamics, safety, and efficacy of recombinant factor VIIa in Japanese patients with hemophilia with inhibitors. AB - A multicenter and open-labeled clinical trial of human recombinant factor VIIa (rFVIIa) was conducted in Japanese patients with severe hemophilia A or B with inhibitors. The trial consisted of 2 parts. In study 1, the pharmacokinetics, pharmacodynamics, and safety of a single dose of 120 microg/kg of rFVIIa were investigated in 8 patients. In the subsequent study 2, the hemostatic effect and safety of rFVIIa were evaluated during a 24-week period in 10 patients. In study 1, the mean maximum FVII-coagulant activity (FVII:C) was found to occur after 10 minutes; activity then decreased rapidly and returned to the baseline within 24 hours after a single intravenous infusion of rFVIIa. The mean half-life of FVII:C was 3.5 hours. The activated partial thromboplastin time and prothrombin time in the patients were immediately shortened but returned to the baseline within 24 hours after dosing. In study 2, 86 microg/kg to 120 microg/kg of rFVIIa (mean, 97 microg/kg) was administered 1 to 85 times to 10 patients. A total of 58.0% (91/157) of bleeding episodes were treated excellently or effectively, with 5 (3.2%) ineffective episodes. There was no apparent trend in the relationship of the hemostatic effect with bleeding sites, mean dose, or number of injections. The efficacy rate, however, was significantly higher (90.0%) in bleeding episodes treated within 3 hours than in those treated at longer intervals (31.0%). No treatment-related adverse events were observed, and there was no evidence of antibody formation to rFVIIa. In conclusion. rFVIIa is an effective and well tolerated option for treatment of bleeding episodes in hemophilia patients with inhibitors. PMID- 11503969 TI - Lupus anticoagulant-like activity observed in a dimeric lambda protein produced by myeloma cells. AB - We report here a lupus anticoagulant (LA)-like activity observed in a 45-year-old man with Bence-Jones protein (BJP) lambda-type multiple myeloma. This patient showed no clinical symptoms of thrombosis or bleeding diathesis. Laboratory examination on admission showed mild anemia, prolongation of activated partial thromboplastin time (APTT) (APTT, 56.2 seconds; control, 29.1 seconds), normal prothrombin time, normal thrombin time, and massive proteinuria (2.3 g/d). The mix test with normal plasma showed the presence of circulating anticoagulant. Based on the assumption that the lambda-type BJP may have been responsible for the prolongation of APTT, we purified the BJP from the patient's urine using column works. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting showed that the purified protein was a 48-kd homodimer of immunoglobulin lambda-chains. Addition of the purified dimeric lambda-type BJP to the normal plasma prolonged both APTT and dilute Russell's viper venom time (DRVVT) in a dose-dependent manner, and the negatively charged phospholipid dependent prothrombinase activity was significantly inhibited in the presence of this protein. Furthermore, both the prolongation of DRVVT and the inhibition of the prothrombinase activity were almost completely abrogated under the condition of high ionic strength. These findings collectively suggest that the dimeric lambda-type BJP showed LA-like activity via the mechanism of ionic charge. PMID- 11503970 TI - Chronic idiopathic thrombocytic purpura in a young male patient with isolated IgM deficiency. PMID- 11503971 TI - Two-dimensional gel electrophoresis of nuclear proteins of different stages of B chronic lymphocytic leukaemia. AB - Two-dimensional polyacrylamide gel electrophoresis was used to compare the composition of nuclear proteins from normal and B-chronic lymphocytic leukaemia (B-CLL) mononuclear cells. Some differences in the electrophoretic behaviour of these proteins from normal and transformed cells, especially with molecular weights/pI of 14-16 kD/5.9-7.4; 28-32 kD/4.9-5.5; 38-39 kD/5.4-6.1; 44-46 kD/5.1 5.6; 47-52 kD/5.0-5.6; 64-69 kD/5.1-5.7, and 95-105 kD/5.2-5.5, were observed. The comparative analysis of nuclear proteins, obtained from mononuclear cells of patients with B-CLL at different stages of development, indicated that the expression of some protein components might be correlated with the progression of this disease. PMID- 11503972 TI - Morphohistobiochemical characteristics of embryogenic and nonembryogenic callus cultures of sweet potato (Ipomoea batatas L.). AB - Ipomoea batatas callus culture raised in a medium supplemented with 2,4-D (2,4 dichlorophenoxy acetic acid) alone or 2,4-D in combination with benzyl adenine, were found to be embryogenic. Supplementation of exogenous chemicals, such as 5 g/l NaCI or 0.7 g/l proline together with a mild dose of 0.2 mg/l 2,4-D, enhanced somatic embryogenesis significantly in all the genotypes tested. Morphological, growth, physiological, histological, and biochemical characteristics of the embryogenic callus were different from the nonembryogenic callus. The former was compact, slow growing, and nodular compared with the fast growing, fragile, nonembryogenic callus. The embryogenic callus tissue had more dry matter, protein and reducing sugar contents compared with the less embryogenic callus. The somatic embryogenic response remained steady in the cultures for up to 96 weeks. PMID- 11503973 TI - Evaluation of fitness components in strains of Drosophila mulleri carrying different genotypes for an esterase. AB - Several fitness components in strains of Drosophila mulleri carrying the slow or the fast alleles for the major beta esterase (esterase-4) found in this species, as well as in heterozygous flies in which the slow or fast alleles came from one of the parents, were evaluated. Twelve components were analysed including longevity of both virgins and mated males and females, productivity, viability, including the egg-larva, egg-pupa, egg-imago and pupa-imago periods. These parameters were used to estimate the total fitness for each genotype. The best score was reached by individuals having the Est-4S/Est-4S genotype (scored at 1.000), followed by a fitness value of 0.892 presented by the Est-4F/Est-4S genotype (with the fast allele from maternal origin), 0.863 for the Est-4F/Est-4F and 0.842 for the Est-4S/Est-4Fgenotypes (with Est-4F of maternal origin). These results suggested a higher relative adaptability of the Est-4S/Est-4S genotype followed by the Est-4F/Est-4S hybrid that possessed the allele Est-4S of maternal origin, which was incompatible with predictions of neutral polymorphism. PMID- 11503974 TI - Protective effect of poly I:poly C from gentamicin nephrotoxicity in guinea pigs. AB - Poly(inosinic) and poly(cytidylic) acids (Poly I:Poly C) have been used to induce the production of endogenous interferon or release preformed interferon in mammals. Interferon increases the resistance of the cells. Sixty guinea pigs were used to investigate whether Poly I:Poly C gave protection from gentamicin nephrotoxicity. The animals were divided into six equal groups. Group 1 were controls; group 2 received gentamicin intramuscularly; group 3 received gentamicin and 12 h later frusemide; group 4 received gentamicin and 12 h later 1 deamino-8-D-argine vasopressin (DDAVP) intramuscularly; group 5 received subcutaneously Poly I:Poly C; group 6 received Poly I:Poly C and 24 h later gentamicin. Frusemide in group 3 potentiated gentamicin nephrotoxicity while DDAVP in group 4 ameliorated gentamicin nephrotoxicity. Poly I:Poly C itself had no toxic effect on renal tissue, while Poly I:Poly C followed 24 h later by gentamicin indicated a protective effect from the gentamicin nephrotoxicity as the functional and histological investigations indicated. PMID- 11503975 TI - Anticlastogenic effects of d- and l-centchroman in Swiss albino mice. 2. Subacute study in vivo and comparison with tamoxifen. AB - The antimutagenic effects of the two enantiomers of centchroman, a nonsteroidal oral contraceptive, were evaluated and compared with tamoxifen, a known breast cancer drug. Anticlastogenic assays in subacute in vivo studies in Swiss albino mice were used. They revealed that both d-centchroman and I-centchroman reduced the chromosome aberrations produced by dimethylbenz(a)anthracene and cyclophosphamide, when compared with the group treated only with the former mutagen. Tamoxifen also reduced the chromosome aberrations produced by the two mutagens. Overall the results showed that l-centchroman alone was more effective in reducing cyclophosphamide-induced aberrations than d-centchroman, and for toxicity reasons may be an alternative to tamoxifen in breast cancer therapy. PMID- 11503977 TI - Zen and the art of surgery. PMID- 11503976 TI - Correlation between the expression of cyclin A protein and p53 activity in oral squamous cell carcinomas. AB - Cyclins and wild-type p53 protein are prime cell cycle regulators and may be involved in tumorigenesis. Cyclin A is a late S cyclin and its abnormalities have been reported in several cancers, including oral squamous cell carcinomas. To explore whether aberrant G1/S in p53 mutant tumours leads to increased cyclin A protein in oral squamous cell carcinomas (OSCC), a total of 39 samples were evaluated for the expression of cyclin A and p53 protein by an immunohistochemical method using a labelled polymer assay. These samples comprised two hyperkeratotic and three oral premalignant lesions (two moderate and one severe dysplastic lesions), and 27 OSCC, together with seven healthy controls. The results demonstrated that the cyclin A protein was localized and highly expressed in the nuclei of the tumour cells. Although there was no correlation between cyclin A detection and the local lymph node involvement, a positive correlation was noted between the positivity of cyclin A and p53 protein (p <0.05). The results suggested that cyclin A may contribute to the progression of oral cancer and correlated to some degree with that of the p53 gene activity. PMID- 11503978 TI - Arthroscopically assisted autologous osteochondral transplantation for osteochondral lesions of the talar dome: an MRI and clinical follow-up study. AB - Osteochondral Lesions of the Talar Dome (OLT) are common problems encountered in orthopedics. Although the etiology remains uncertain, a myriad of treatment options exists. The authors describe arthroscopically assisted autologous osteochondral graft (OCG) transplantation procedures in the treatment of unstable OLTs in nine patients. The patients underwent standard preoperative MRI examination to assess fragment stability (using De Smet criteria for stability). Intraoperative arthroscopy was used to correlate the preoperative MRI assessment (using Cheng/Ferkel grading). After transplantation procedures, MRI (using De Smet criteria for stability) assessed graft incorporation for stability at an average of 9.3 months after the procedure. Preoperative MRI correlated highly with arthroscopic findings of OLT instability (sensitivity = 1.0). This has been demonstrated in the current orthopedic literature. The post transplantation MRI demonstrated stable graft osteointegration by De Smet criteria in all patients. Postoperative visual analogue pain scales showed significant improvement from preoperative assessment. Postoperative AOFAS Ankle-Hindfoot scores averaged 80.2 (S.D. +/- 18.9). Our favorable early results and those of other authors using similar techniques may validate OCG transplantation as a viable alternative for treating unstable osteochondral defects in the talus that are refractive to more commonly used surgical techniques. PMID- 11503979 TI - Mosaicplasty for the treatment of osteochondritis dissecans of the talus: two to seven year results in 36 patients. AB - An osteochondral defect (OCD) is known as a symptomatic lesion causing pain, recurrent synovitis, and altered joint mechanics most commonly in a weight bearing joint. Loose bodies may develop, which may then cause joint destruction and/or locking. The damage to the articular surface is most likely a precursor of ankle osteoarthritis. With the recent advances in diagnostic imaging, such as MRI, as well as the development of ankle arthroscopy, the identification and classification of these lesions has become much more precise. This allows more accurate staging and improves treatment recommendations. The assessment of a particular treatment is also improved. A variety of treatment alternatives are now available. These include arthroscopic procedures including debridement, retrograde drilling, and bone grafting. Compared to open treatment, arthroscopic procedures may be particularly advantageous in the treatment of small defects and stable OCD lesions. Until recently, however, favorable results have been less predictable for large or unstable osteochondral defects. We treat these more difficult lesions with a mosaic autogenous osteochondral transplantation. In our hands, this appears to provide an optimal treatment result. The present report evaluates the clinical outcome of 36 patients followed for two to seven years after a mosaicplasty autogenous osteochondral transplantation from a non or less weight bearing portion of the knee to the ipsilateral talus. Ankle function was measured by the Hannover scoring system and showed good to excellent results in 34 cases (94%) with no long term donor site morbidity. The encouraging clinical results are supplemented with radiographs and histology, which support the premise of lasting relief of symptoms and prevention of ankle arthrosis. PMID- 11503980 TI - Percutaneous vs. open repair of the ruptured Achilles tendon--a prospective randomized controlled study. AB - A prospective randomized controlled trial comparing open and percutaneous repair of closed ruptured Achilles tendons was performed over a period of 30 months. Sixty-six patients from seven district general hospitals were entered into the study with 33 patients randomized into each group. A modification of the technique described by Ma and Griffith was used in the percutaneous group and a Kessler suture supplemented with interrupted sutures was used in the open group. Patients were followed up for a minimum of six months. The mean age was 38.5 years (26 to 53 years). Forty patients were male and 26 female. After the rupturing event but prior to surgery, it was noted that seven patients had paresthesia in the territory of the sural nerve. The mean duration of immobilization was 12.4 weeks (10 to 14). The complications in the open group included seven wound infections (21%), two adhesions (6%) and two cases of re rupture (6%). In the percutaneous group there were three cases of wound puckering (9%), one re-rupture (3%) and one case with persistent paresthesia in the sural nerve territory (3%). The difference in infective wound complications between the two groups was statistically significant (Fisher's exact test P = 0.01). Percutaneous repair is advocated on the basis of the low rate of complications and improved cosmetic appearance. PMID- 11503981 TI - Anatomic study of the medial neurovascular structures in relation to calcaneal osteotomy. AB - Medial displacement osteotomy of the calcaneous is commonly performed for stage II posterior tibial tendon insufficiency in an effort to improve the valgus deformity of the hindfoot. We performed an anatomic study examining the medial neurovascular anatomy and its relation to the osteotomy in an attempt to determine which structures may be at risk during the procedure. Calcaneal osteotomies were performed through a lateral approach on 22 fresh-frozen cadaver below-knee specimens. Dissection was then performed medially to identify the Medial Plantar Nerve (MPN), the Lateral Plantar Nerve (LPN), the Posterior Tibial Artery (PTA), and their respective branches. Measurements determined either 1) where the structure crossed the osteotomy or 2) if the structure did not cross, the closest perpendicular distance from the osteotomy and at which point along its length this occurred. Perpendicular distances were recorded in millimeters and position along the osteotomy as a percentage of the total length from the posterosuperior aspect. An average of four neurovascular structures crossed each osteotomy site (range 2 to 6), most of which were branches of the LPN or the PTA. The MPN did not cross in any of the specimens studied, the LPN crossed in one specimen, and the PTA crossed in two specimens. The MPN distributed no crossing branches. The calcaneal sensory branch of the LPN was identified and crossed in 86% of the cadavers at 19% (+/- 15%) along the osteotomy length. A more distal second branch of the LPN (Baxter's nerve) was identified and crossed in 95% of the specimens at 61% (+/ 20%) along the osteotomy length. A third crossing branch existed in one specimen. Each PTA distributed from zero to three branches which variably crossed the osteotomy at a point from 2% to 100% along its length. The PTA bifurcated in 77% of the specimens at 49% (+/- 9%) along the osteotomy length. A consistent finding in every specimen was the presence of two veins accompanying the PTA with one on either side. A number of medial neurovascular structures may be at risk when perfoming a calcaneal osteotomy through a lateral approach. A minimum of two structures crossed the osteotomy site at variable positions in this study, although most of these structures represented branches off of the LPN or the PTA, with the LPN and the PTA themselves crossing only infrequently. The authors recommend that the completion of the osteotomy through the medial calcaneal cortex be performed in a carefully controlled manner to reduce the risk of post-operative complications including pain, numbness, and hematoma formation. PMID- 11503982 TI - An analysis of skin perfusion over the achilles tendon in varying degrees of plantarflexion. AB - Delayed wound healing and, less commonly, wound breakdown are significant complications following open Achilles tendon repair. Skin perfusion over the Achilles tendon may be reduced when the ankle is plantarflexed. The aim of this study was to analyse skin perfusion over the Achilles tendon with the ankle in varying degrees of plantarflexion in 20 volunteers. Skin perfusion was determined by measuring the transcutaneous skin oxygen pressure (tcPO2) using the Novametrix TcO2M 860 monitor. Measurements were taken at the medial edge of the Achilles tendon in 20 volunteers. Skin perfusion was maximal with the ankle plantarflexed to 20 degrees. With plantarflexion beyond this skin perfusion was reduced. At 40 degrees plantarflexion skin perfusion was reduced by up to 49% (mean 35%, range 27% to 49%). We conclude plantarflexion beyond 20 degrees reduces skin perfusion in the region of the Achilles tendon. Though this study was performed on non operated cases, and is thus limited, the findings may have clinical implications with regard to cast position following Achilles tendon repair. PMID- 11503983 TI - Biomechanical comparison of ankle arthrodesis techniques: crossed screws vs. blade plate. AB - Many different techniques for ankle arthrodesis have been described. Experience at our institution with crossed screws internal fixation has not met the 90+% union rate reported in the literature. A compression blade plate is one technique for ankle arthrodesis which has not been evaluated biomechanically. A biomechanical study comparing two groups of sawbone ankle fusion constructs fixed with crossed screws and compression blade plates was performed in order to evaluate the stiffness and rigidity of these two arthrodesis techniques. The crossed screws construct demonstrated superior stiffness during dorsiflexion (p < 0.001) and valgus (p < 0.001) loading. The two constructs were found to have equal strength in resisting plantarflexion, varus and torsional loads although there was a trend for greater resistance by the crossed screws construct. These findings lend biomechanical support to the use of crossed screws for tibiotalar arthrodesis. PMID- 11503984 TI - Intramedullary screw fixation of proximal fifth metatarsal fractures: a biomechanical study. AB - Intramedullary screw fixation is a popular technique for treatment of proximal fifth metatarsal fractures. The purpose of this study was to compare the fixation rigidity of a 5.5 mm partially threaded cannulated titanium screw, with presumed superior endosteal purchase, to a similar 4.5 mm screw. Acute fifth metatarsal fractures were simulated in cadavers, stabilized with intramedullary screws, and loaded to failure in three-point bending. The initial failure loads for the metatarsals fixed with 4.5 mm and 5.5 mm screws were not significantly different (332.4 N vs. 335.2 N, respectively), nor were the ultimate failure loads (849.8 N vs. 702.2 N, respectively). Based upon our results, maximizing screw diameter does not appear to be critical for fixation rigidity and may increase the risk of intraoperative or postoperative fracture. PMID- 11503986 TI - Effect of distal fibular resection on ankle laxity. AB - Ten cadaver specimens were used to study the effect of distal fibular resection and ligamentous repair on ankle laxity. The distal fibula was resected sequentially at three levels. Talar tilt and anterior drawer were measured on stress radiographs after resection and then after ligament repair. Sequential resection of the distal fibula caused a progressive increase in talar tilt and anterior drawer at each cut level (p < 0.5), regardless of whether the ligaments were repaired or not. Ligament repair significantly decreased talar tilt at cuts above the ATFL insertion and decreased anterior drawer at cuts at the tibiotalar level, but these values did not approach control values. PMID- 11503985 TI - Intramedullary screw fixation of Jones fractures. AB - BACKGROUND: Jones fractures of the fifth metatarsal can be stabilized using intramedullary screw fixation techniques. A range of screw diameters from 4.5 mm to 6.5 mm can be used, but the optimal screw for this procedure has yet to be defined. In clinical practice, we have observed that failure is more likely when smaller diameter screws are used. METHODS: Experimental Jones fractures were created in 23 pairs of human cadaver fifth metatarsals, which were fixed using either 5.0 mm or 6.5 mm screws. Fracture stiffness and pull-out strengths were measured for either screw type and their relationships with bone mineral density and medullary canal diameter were determined. RESULTS: There was no significant difference in the bending stiffness of fractures stabilized with 5.0 mm and 6.5 mm screws; however, different mechanisms of failure were noted for either screw type. Poor thread purchase within the medullary canal was noted with the 5.0 mm screws, while excellent purchase was noted with 6.5 mm screws. Pull-out strength testing revealed significantly higher pullout strengths for the larger 6.5 mm screws. There was no significant difference in bone mineral density or medullary canal diameter between right and left metatarsals. CONCLUSIONS: Fifth metatarsals can often accommodate a 6.5 mm screw for the stabilization of Jones fractures. Larger diameter screws did not result in greater fracture stiffness in our model, but did result in significantly greater pull-out strengths. CLINICAL RELEVANCE: Larger diameter screws may be more appropriate for intramedullary screw fixation of Jones fractures. PMID- 11503987 TI - Foot pressures during gait: a comparison of techniques for reducing pressure points. AB - PURPOSE: Various methods have been used to redistribute plantar surface foot pressure in patients with foot ulcers. This study was conducted to determine the effectiveness of four modalities (fracture walker, fracture walker with insert, and open and closed toe total contact casts) in reducing plantar foot pressure. METHODS: Ten healthy, normal volunteer subjects had an F-scan sensor (ultra thin shoe insert pressure monitor) placed under the right foot. They then ambulated on a flat surface, maintaining their normal gait. Dynamic plantar pressures were averaged over 10 steps at four different sites (plantar surface of great toe, first metatarsal head, base of fifth metatarsal, and plantar heel). All subjects repeated this sequence under five different testing conditions (barefoot, with a fracture walker, fracture walker with arch support insert, open and closed toe total contact cast). Each subject's barefoot pressures were then compared with the pressures during the different modalities. RESULTS: All four treatment modalities significantly reduced (p < 0.05) plantar pressure at the first metatarsal head (no method was superior). The fracture walker, fracture walker with insert, and open toe total contact cast significantly reduced pressure at the heel. Pressures at the base of the fifth metatarsal and great toe were not significantly reduced with any treatment form. CONCLUSION: The fracture walker, with and without arch support, and total contact cast can effectively reduce plantar pressure at the heel and first metatarsal head. PMID- 11503988 TI - The effect of shoe sole composition on in vivo tibial strains during walking. AB - To test the hypothesis that shoe sole composition can affect the level of bone strain and strain rates that contribute to the development of stress fractures, in vivo tibial strain measurements were made during treadmill walking while wearing four shoes which differed only in their sole composition. Soles of 65 Shore A polyurethane with an embedded heel air cell had significantly lower compression and shear strains and shear strain rates than soles of 65, 75 and a composite of 40/65 Shore A polyurethane with no embedded air cells. A sole composed of 65 Shore A polyurethane with an embedded air cell can potentially be protective against stress fracture in a walking shoe. PMID- 11503989 TI - Proximal fourth metatarsal injuries in athletes: similarity to proximal fifth metatarsal injury. AB - Proximal fourth metatarsal injuries are rarely reported. We present five case histories in which athletic patients sustained injuries at the shaft-base junction of the fourth metatarsal. Similar to proximal fifth metatarsal injuries, adduction of the forefoot appears to be associated. Our patients returned to their activities in two to eight months. These patients injuries tended to take longer to heal than other lesser metatarsal fractures and stress fractures (which are typically more distal). Some patients were continually symptomatic, even after three months of rest and immobilization. This coincides with proximal fifth metatarsal injuries and stress fractures. For treatment of proximal fourth metatarsal injuries to be successful, ideal treatment appears to involve nonweightbearing below-knee cast/boot immobilization for three weeks. This is followed by an additional three or more weeks of weightbearing immobilization. Healing may still be prolonged. PMID- 11503990 TI - Anterior dislocation of the subtalar joint: a case report. AB - We report a very rare case of anterior dislocation of the subtalar joint. Forceful supination of the foot and dorsiflexion of the ankle was considered the cause of the injury in this case. Closed reduction was successful for the talocalcaneal component of subtalar joint, although surgery was subsequently performed because of the residual subluxation of the midtarsal joint including the talonavicular component of subtalar joint and the associated fracture of the lateral process of the talus. Satisfactory results were shown at three-year follow-up. PMID- 11503991 TI - Sealing ability of Dyract, Geristore, IRM, and super-EBA as root-end filling materials. AB - The purpose of this study was to evaluate the apical sealing ability of two compomers (Dyract and Geristore), IRM, and Super-EBA. Forty single canal roots from human teeth were instrumented until a size 40 file extended 1 mm beyond the apex. One millimeter of root apex was removed and a preparation 3-mm deep was prepared. Roots were divided into 4 groups of 10 roots each and filled with IRM, Super-EBA, Dyract, or Geristore. Canals were not obturated to ensure that any leakage was due to the apical filling material alone. Each root was then affixed to a fluid filtration device and subjected to a pressure of 14 cm of H2O, which has been determined to be the normal pulpal tissue pressure. The integrity of the seal was evaluated for 5 min at 1, 7, 30, and 180 days. Data were analyzed at each time point using one-way analysis of variance on ranks. The results of this study suggest that the new compomers Dyract and Geristore are equal or superior to IRM and equivalent to Super-EBA in their ability to reduce apical leakage when used as retrofilling materials. PMID- 11503992 TI - Presence of interleukin-4-producing cells for human bone regeneration after application of guided tissue regeneration membranes. AB - To study the process of bone regeneration we examined three samples of periapical regenerative tissue obtained from two patients under a guided tissue regeneration treatment in endodontic surgery by the immunohistochemical and enzyme histochemical methods. The regenerative tissue consisted of a large number of fibroblast-like cells and a small number of mononuclear cells. Fibroblast-like cells stained positively for alkaline phosphatase and osteopontin, whereas mononuclear cells stained positively for CD4. Interleukin-4-producing cells could be detected in adjacent sections. However, interferon-y-producing cells could not be detected. These findings suggest that interleukin-4-producing cells may be one of the elements associated with success in the human bone regeneration process in vivo. PMID- 11503993 TI - In vitro apical leakage of system B compared with other filling techniques. AB - A fluid filtration system was used to compare the apical microleakage of roots filled with the system B (Analytic Technology), single-cone technique, lateral condensation, vertical condensation, and Thermafil (Caulk-Dentsply). After preparation of the canal system with a Profile (Caulk-Dentsply) five groups of 10 single-rooted teeth were randomly filled according to 1 of the 5 techniques. Phosphate-buffered saline was forced under a 15 cm H2O pressure, through the apex, toward the coronal part of the root. The filtration rate in L s(-1) Kpa(-1) was recorded 24 h after filling and after 1-month storage in phosphate-buffered saline. The 24 h results showed that the single cone technique provided the highest leakage (p = 0.001). At 1-month system B, Thermafil and vertical condensation had less leakage than the two other techniques (p = 0.0001): lateral condensation produced moderate apical leakage, whereas the single cone technique showed the highest leakage. Regardless of the technique the apical leakage increased after 1-month storage (p = 0.001 or p = 0.0001, depending on the technique). PMID- 11503994 TI - In vitro assessment of the antimicrobial action and the mechanical ability of chlorhexidine gel as an endodontic irrigant. AB - The objective of this study was to assess the chlorhexidine gluconate gel as an endodontic irrigant. First the ability of chlorhexidine gel to disinfect root canals contaminated in vitro with Enterococcus faecalis was investigated. A scanning electron microscope was also used to evaluate its cleansing ability compared with endodontic irrigants commonly used, such as sodium hypochlorite and chlorhexidine gluconate liquid. The results indicated that the chlorhexidine gel produced a cleaner root canal surface and had an antimicrobial ability comparable with that obtained with the other solutions tested. It was concluded that chlorhexidine gluconate in gel form has potential for use as an endodontic irrigant. PMID- 11503996 TI - P53 protein expression in odontogenic cysts. AB - p53 protein seems to be related to the suppression of cell proliferation. p53 positive tissues seem to have a higher proliferative activity than p53-negative ones. Odontogenic keratocyst (OKC) has a different behavior from other types of cysts because it is more aggressive, with a tendency to recurrence. Twenty-two dentigerous cysts, 24 radicular cysts, and 20 OKCs were used in the present study. Two dentigerous cysts (9.1%), 2 radicular cysts (8.3%), and 9 OKCs (45%) expressed the p53 protein. The differences between the three groups were statistically significant (p = 0.003). In 10 cases of OKCs epithelial dysplasia was found. One of the 10 OKCs without dysplasia and 8 of the 10 OKCs with dysplasia were p53-positive: the difference between the two groups was statistically significant (p = 0.007). The overexpression of p53 protein was not on the other hand correlated with the occurrence of multiple, bilateral, and recurrent OKCs. Moreover the distribution of p53-positive cells was parabasal in contrast with other types of cysts. These qualitative and quantitative differences in proliferative activity in OKCs seem to point to an alteration in cell cycle control. PMID- 11503995 TI - Microleakage of root-end filling materials. AB - The purpose of this study was to evaluate the microleakage of various root-end filling materials using a fluid filtration system. Sixty extracted human single rooted teeth were used. The crowns were removed, the canals prepared, and root end fillings placed. The samples were divided into two control and five experimental groups. The root-end filling materials tested were: amalgam, Intermediate Restorative Material (IRM), a dentin-bonded resin, Super-EBA, and mineral trioxide aggregate. The results showed that amalgam root-end fillings demonstrated significantly more microleakage than Super-EBA, dentin-bonded resin, or mineral trioxide aggregate. There was no significant difference between amalgam and IRM. However IRM was also not significantly different from the other three groups. There were no significant differences between the other three groups. PMID- 11503997 TI - Matrix metalloproteinase-2 in dentin matrix mineralization. AB - In the serum-free culture medium of bovine odontoblasts we detected active gelatinolytic metalloproteinases, matrix metalloproteinase (MMP)-2 and MMP-9 (gelatinases A and B). The activity of MMP-2, in particular, appeared suddenly around day 21 in the culture, coinciding with the development of odontoblastic cell processes and the loss of alkaline phosphatase. Reverse transcriptase polymerase chain reaction analysis of these odontoblasts demonstrated that messages of MMP-2 but not MMP-9 increased significantly between day 15 and day 21. The in vitro observation indicates that medium conditioned by these odontoblasts and containing significant amounts of MMP-2 degrades not only the collagenous substrates but also purified dentin phosphophoryn as well. We have also observed that dephosphorylated dentin phosphoprotein becomes a better substrate for casein kinase II after limited proteolysis with MMP-2. These results support our working hypothesis that MMP-2-mediated proteolytic processing is an important step in accelerating the process of dentin matrix maturation, which includes phosphorylation and subsequent mineralization. As has been suggested previously, extracellular phosphorylation of matrix proteins is an important step in biomineralization both in bone and in dentin (Mikuni-Takagaki et al., J Bone Miner Res 1995;10:231-42; Zhu et al., Biochem J 1997; 323:637-43). Our present histochemical analysis in MMP-2 knockout mice confirms the concept with the delayed formation of mineralized tissues, dentin, and bone. PMID- 11503998 TI - Feline immunodeficiency virus model to study human immunodeficiency virus/acquired immune deficiency syndrome conditions. AB - This study was designed to induce rapid progression of the feline immunodeficiency virus (FIV) infection in cats. Predictably inducing the FIV disease state in the cat would yield an excellent tool to study endodontic disease processes under immunosuppressed conditions. Eight cats were immunosuppressed with steroids before infection with FIV. Another eight cats, age and sex-matched littermates, served as uninoculated seronegative controls. Complete blood counts were taken for 10 mo in the FIV group, and 10 wk in the control group, including lymphocyte subsets. ELISAs were used to detect FIV infection. Statistical analysis was performed with generalized estimating equation models. All cats were positive at one point in time. The FIV group had significantly lower peripheral blood CD4+ counts compared with the control group. Therefore the FIV model presented gives the desired outcome and simulates what occurs in human immunodeficiency virus infection. PMID- 11503999 TI - Benchmarking the endodontic literature on MEDLINE. AB - The purpose of this study was to identify and quantify the endodontic literature available for clinical decision making. A search strategy based on Medical Subject Headings for endodontics was developed to examine MEDLINE. The identified articles were limited to human subjects and English. Sensitive and specific methodological search filters identified four categories of information: etiology, diagnosis, therapy, and prognosis. The results were then subdivided by year to identify trends. Between 1990 and 1998 MEDLINE identified 3,152 articles published in English on endodontics in humans. The number of articles per year (mean +/- SD) for sensitive and specific searches was etiology (28+/-10, 1+/-2), diagnosis (38+/-11, 1+/-1), therapy (59+/-15, 3+/-3), and prognosis (40+/-13, 10+/-5), respectively. The number of articles in each category increased by 1 to 3% each year. There were 150 articles/yr in endodontics in at least 120 journals cited on MEDLINE on which to base clinical decisions. PMID- 11504000 TI - Treatment of a mandibular molar with perforating internal resorption. AB - A case is presented in which a previous partial root canal treatment on a mandibular molar developed internal resorption of the distal canal. The case was followed at the University of Oklahoma student clinic and endodontic treatment completed in the mesial canals, with calcium hydroxide placed in the distal canal for 10 months due to a pre-existing perforating defect of the root surface. Endodontics was then completed. A 17-month follow-up film showed osseous repair apically and also adjacent to the distal root surface where a slight overextension of filling material was evident. PMID- 11504001 TI - Aspergillosis of the maxillary sinus as a complication of overfilling root canal material into the sinus: report of two cases. AB - Aspergillosis of the maxillary sinus is a relatively rare disease in nonimmunocompromised patients. In recent years a number of cases of aspergillosis of the maxillary sinus have been reported in association with overextension of root canals fillings with certain root canal cements. It has been suggested that zinc oxide-based root canal cements might promote the infection with the Aspergillus species. In particular Aspergillus fumigatus has been found to be associated with the maxillary sinus infection. Radiographically the unique appearance of a dense opacity foreign body reaction in the maxillary sinus was considered a characteristic finding in maxillary sinus aspergillosis. Because this association of overfilling of root canal cements and aspergillosis of the maxillary sinus is not too well known we report two cases of young healthy female patients with the characteristic findings, both radiographically and clinically. In both patients the first maxillary molar was involved. Patients were symptomless and the diagnosis was made accidently. However at surgical inspection both patients revealed aspergillomas, including the overextended root canal cement. The surgical procedure is described as are the microscopic findings in both cases showing the characteristic branching hyphae and conidophores typical of Aspergillus. Overextension into the maxillary sinus with root canal cements has to be avoided; material has to be removed from the sinus because otherwise aspergillosis infection may ensue. PMID- 11504002 TI - Zebra XXI, part 1. PMID- 11504003 TI - Similarity, global matching, and judgments of frequency. AB - In the test-pair similarity effect, forced-choice recognition is more accurate for similar test pairs, such as leopard-cheetah, than it is for unrelated test pairs, such as leopard-turnip. According to global matching models, this occurs because the retrieved familiarities of similar items are correlated. In the Minerva 2 model, global matching underlies frequency judgments as well as recognition memory. One implication of this model is that judged frequencies of similar items should be correlated. Another implication is that judgments of summed frequency for pairs of words (how many presentations were there of word1 and word2 combined?) should have higher variance when word1 and word2 are similar than when they are unrelated. These predictions were tested and confirmed in two experiments. A review of these and other results suggests that theories of recognition memory should also be applicable to frequency-judgment tasks. PMID- 11504004 TI - Effects of word length on young children's memory performance. AB - The standard working model that accounts for word length effects in young children has been questioned. This study explored two hypotheses suggested by Gathercole and Hitch (1993). In three experiments with serial recall tasks, the word presentation interval was adjusted so that participants could repeat both long and short words subvocally the same number of times in the intervals between the words presented. As a result, among children from 3 to 6 years of age, the word length effect was significantly reduced, whereas the same manipulation of the word presentation interval did not affect the word length effect in adults. These results suggest that the word length effects in young children reflect the process of retaining auditory information in the interval between presentations as well as readout of the phonological representation. PMID- 11504005 TI - Structural alignment facilitates the noticing of differences. AB - High-similarity concept pairs that elicit many commonalities also elicit many related differences (Gentner & Markman, 1994; A. B. Markman & Gentner, 1993a, 1993b, 1996; A. B. Markman & Wisniewski, 1997). This finding has been used to support the claim that the comparison process is one of structural alignment. However, it is possible that the difference advantage results from some other property of high-similarity pairs, such as a greater number of stored differences. The present experiments demonstrate that the comparison process itself leads to the greater psychological availability of differences. In three experiments, participants listed commonalities for word pairs and then listed differences under a time pressure for these old pairs and new pairs. In Experiment 1, participants listed more differences for old than for new pairs, consistent with the claim that the comparison process facilitates noticing differences. In Experiment 2, we showed that the difference-listing advantage is specific to the comparison process: Mere coprocessing of the pairs (specifically, providing thematic relations) does not facilitate, and in fact appears to inhibit, difference listing. In Experiment 3, pairs with deeper common systems elicited a larger number of specific alignable differences than did pairs with shallow sets of commonalities. Overall, the results support the structural alignment claim that the comparison process promotes the noticing of both commonalities and related differences. PMID- 11504006 TI - A group size effect on personal risk judgments: implications for unrealistic optimism. AB - In Experiments 1 and 2, college students (N = 32 and N = 18, respectively) read heart attack risk profiles (i.e., lists of risk factors) for each of several employees at a series of fictional companies and judged the heart attack risk of the typical employee at each company. In both experiments, subjects' risk judgments increased as a function of the number of employees at the companies. In Experiments 3A and 3B, college students (N = 56 and N = 33, respectively) judged the heart attack risk of the typical employee at a company and also judged the risk of each individual employee. In these experiments, the typical employee was generally judged to be at higher risk than the individual employees. This group size effect might help to explain unrealistic optimism--people's tendency to judge themselves to be at lower risk than their peers for negative life events. Furthermore, it can be modeled successfully within Fiedler's (1996) BIAS framework. PMID- 11504007 TI - An inverse base rate effect with continuously valued stimuli. AB - It is well known that people do not always make normative use of information about relative frequencies of categories when making categorical judgments. The "inverse base rate" effect (Medin & Edelson, 1988) is a typical example of this: Subjects violate normative reasoning principles by assigning certain ambiguous stimuli as belonging to the less frequent of two categories, rather than to the more common category. This effect has been explained as being due to the shifting of attention from shared stimulus features to distinctive features during learning. When stimuli are defined by values along continuous dimensions, rather than by the presence and absence of features, then attention could shift between dimensions or between values, or both. In three experiments, base rate differences were used to determine the way in which attention is shifted during learning about stimuli with continuously valued dimensions. Simulation modeling shows that the results are consistent with the movement of attention both between and within stimulus dimensions. PMID- 11504008 TI - Feedback effects on cost-benefit learning in perceptual categorization. AB - Two experiments were conducted in which the effects of different feedback displays on decision criterion learning were examined in a perceptual categorization task with unequal cost-benefits. In Experiment 1, immediate versus delayed feedback was combined factorially with objective versus optimal classifier feedback. Immediate versus delayed feedback had no effect. Performance improved significantly over blocks with optimal classifier feedback and remained relatively stable with objective feedback. Experiment 2 used a within-subjects design that allowed a test of model-based instantiations of the flat-maxima (von Winterfeldt & Edwards, 1982) and competition between reward and accuracy (Maddox & Bohil, 1998a) hypotheses in isolation and of a hybrid model that incorporated assumptions from both hypotheses. The model-based analyses indicated that the flat-maxima model provided a good description of early learning but that the assumptions of the hybrid model were necessary to account for later learning. An examination of the hybrid model parameters indicated that the emphasis placed on accuracy maximization generally declined with experience for optimal classifier feedback but remained high, and fairly constant for objective classifier feedback. Implications for cost-benefit training are discussed. PMID- 11504009 TI - Contextual overlap and eyewitness suggestibility. AB - Studies of eyewitness suggestibility have traditionally used a paradigm that maximizes the extent to which the postevent interview overlaps with the witnessed event in terms of narrative content, narrative structure, and environmental context. The present study explored whether these dimensions of overlap contribute to people's tendency to confuse suggested details for those they have actually witnessed. We systematically manipulated the extent to which the postevent questionnaire overlapped with the witnessed event. Across two experiments, overlap in narrative content, narrative structure, or environmental context was not found to increase suggestibility effects, even though the manipulation did have other memory effects (e.g., it improved cued recall of the actual source of the suggestions, Experiment 2). These findings suggest that understanding the interaction between the structure and content of the objective context in which misinformation is encountered and various remembering contexts (e.g., recognition vs. recall) is important for advancing our understanding of source confusion in an eyewitness situation. PMID- 11504010 TI - Principle explanation and strategic schema abstraction in problem solving. AB - This study was undertaken to examine the effects of strategic schema-acquisition tasks (problem comparison or problem construction) and the method of principle explanation (abstract or embedded principle method) on schema acquisition. Ninety eight subjects studied a set of problems in probability, presented according to either method of principle explanation. Half the subjects in each principle explanation group were then asked to compare analogous problems, and the rest constructed new analogous problems. To determine whether subjects generalized problem schemas, they were given new analogous problems to solve. The results showed that when the abstract principle method was used, schema acquisition was better in problem comparison; but with the embedded principle method, schema acquisition was better in problem construction. Results were discussed in relation to the importance of some fit between the presentation of problem information and the processes that will draw from or build on this information in tasks designed to allow novice problem solvers to acquire advanced problem representations. PMID- 11504011 TI - Taxi drivers' exceptional memory of street names. AB - In this paper, we explore the nature of taxi drivers' serial recall of street names. The main question is whether the memory of verbal material benefits from the possibility of using visuospatial associations and knowledge concerning large scale environment. In two experiments, expert taxi drivers' recall of street names was superior to that of control groups. In Experiment 1, experts' superiority of memory was greater when the street names reflected a visuospatially continuous route than when the street names were located along a straight line across the map without spatial continuity or were presented in random order. In Experiment 2, the expert taxi drivers recalled spatially continuously organized lists much better than they recalled lists of street names belonging to the same semantic category or lists presented in alphabetical order. This result also suggests that interitem associations, which are based on spatial co-occurrence, are efficient in comparison with other mnemonics. PMID- 11504012 TI - Subjective frequency estimates for 2,938 monosyllabic words. AB - Subjective frequency estimates for large sample of monosyllabic English words were collected from 574 young adults (undergraduate students) and from a separate group of 1,590 adults of varying ages and educational backgrounds. Estimates from the latter group were collected via the internet. In addition, 90 healthy older adults provided estimates for a random sample of 480 of these words. All groups rated words with respect to the estimated frequency of encounters of each word on a 7-point scale, ranging from never encountered to encountered several times a day. The young and older groups also rated each word with respect to the frequency of encounters in different perceptual domains (e.g., reading, hearing, writing, or speaking). The results of regression analyses indicated that objective log frequency and meaningfulness accounted for most of the variance in subjective frequency estimates, whereas neighborhood size accounted for the least amount of variance in the ratings. The predictive power of log frequency and meaningfulness were dependent on the level of subjective frequency estimates. Meaningfulness was a better predictor of subjective frequency for uncommon words, whereas log frequency was a better predictor of subjective frequency for common words. Our discussion focuses on the utility of subjective frequency estimates compared with other estimates of familiarity. The raw subjective frequency data for all words are available at http://www.artsci.wustl.edu/dbalota/labpub.html. PMID- 11504013 TI - Local strategic control of information in visual word recognition. AB - In the present studies, we examined strategic flexibility in the use of lexical and sublexical information in Korean word recognition. Korean readers show a large frequency effect for words printed in the alphabetic Hangul script only if these words are embedded in a list consisting largely of logographic (Hanza) words. In the first experiment, high- and low- frequency Hangul words were preceded by Hangul or Hanza words. Frequency effects were eliminated when a Hangul target word was preceded by two Hangul words, even when the overall proportion of Hanza words in the list was large. In the second experiment, one group saw each stimulus preceded by a cue indicating which script would be seen on that trial. Frequency effects for Hangul targets were absent under this condition, but were present when subjects were not so cued. These results indicate that Korean readers are able to control their use of lexical and sublexical information over a small number of stimuli or even trial by trial. PMID- 11504014 TI - Variable diets and changing taste in plant-insect relationships. AB - The host ranges of phytophagous insects are determined to a large degree by plant chemistry. Specialist insects are often closely associated with plants that produce characteristic chemicals, which may act as attractants or stimulants to aid in finding or recognizing a host. Generalist insects are generally believed to rely on the presence of repellents or deterrents to ensure avoidance of unsuitable plants. However, the chemistry of any plant can be highly variable, as a result of growth characteristics, genetic variation, or environmental factors. Such variable chemistry may provide windows of opportunity for nonadapted insects to utilize a plant or for a plant to become resistant to a normally adapted herbivore. Differences in insect responses to plant constituents may also result from genetic variation or environmental factors. In particular, dietary experience has been found to influence the ability of insects to taste plant chemicals that may serve as signals of suitability or unsuitability. Certain dietary constituents appear to suppress the development of taste sensitivity to deterrents in an insect, whereas the presence of specific stimulants in the diet may result in the development of dependence on these compounds. These findings further emphasize the fact that the dynamics of plant biochemistry along with plasticity in the sensory system of insects might be expected to play a major role in the evolution of new plant-insect relationships. PMID- 11504015 TI - Does excretion of secondary metabolites always involve a measurable metabolic cost? Fate of plant antifeedant salicin in common brushtail possum, Trichosurus vulpecula. AB - Salicin was administered orally to six brushtail possums by incorporation in food for six days at three dose levels (0.05, 0.5, and 1.5% wet weight), giving mean +/- SD daily intakes of 0.31 +/- 0.09, 2.76 +/- 0.75, and 6.04 +/- 1.12 mmol salicin. Metabolites were identified by mass spectrometry and assayed by HPLC. Salicyl alcohol glucuronide accounted for 56-64% of urinary metabolites over the three doses, salicyluric acid 15-26%, salicin 10-18%, and there were smaller amounts of free (2-4%) and conjugated (0-6%) salicylic acid. beta,2 Dihydroxyphenylpropionic acid was a minor metabolite. The hydrolysis of dietary salicin enabled reconjugation of its aglycone, salicyl alcohol, with a more polar sugar, glucuronic acid, thus enhancing its renal excretion and resulting in little net loss of substrates for conjugation and a low measurable metabolic cost of excretion. PMID- 11504016 TI - Behavioral and electrophysiological responses of Arhopalus tristis to burnt pine and other stimuli. AB - The exotic longhorn beetle Arhopalus tristis is a pest of pines, particularly those damaged by fire, and a major export quarantine issue in New Zealand. Actinograph recordings of caged individuals showed that males and females were most active from dusk to midnight. Olfactometer experiments indicated that females moved upwind toward odors from burnt pine (80%, N = 75), compared to unburnt pine (20%). Oviposition choice tests showed that eggs were predominantly laid on burnt logs (79%, N = 20), compared to unburnt logs. Beetles were trapped by funnel traps baited with burnt (mean catch per trap 7.8) and unburnt (mean catch 4.1 per trap) pine bark from inside a screen cage (4 x 3 m), while unbaited traps had a mean catch 0.1 beetles (N = 8 replicates). The treatment of burnt pine bark with a 1:1:2 mixture of green leaf volatiles (E)-2-hexen-1-ol and (E)-2 hexenal) in mineral oil as a repellent reduced trap catch by fivefold in a similar experiment (mean catches of 1.2 beetles per trap to burnt pine bark plus repellent treatment and 6.2 beetles per trap to burnt pine bark alone). The treatment of burnt pine bark with this solution also reduced oviposition by 98.5% (mean eggs per log of 11.1 on burnt pine and 0.3 on burnt pine plus repellent), indicating that oviposition cues have the potential to be significantly disrupted. The electrophysiological responses of adult beetles were recorded to a range of odorants. Normalized responses to monoterpenes known to occur in Pinus radiata ranged from about 20 to about 150, with alpha-terpineol giving the greatest responses in both sexes. Green leaf volatiles also gave high responses. The potential exists to improve the management of this insect using chemical cues in various ways. PMID- 11504017 TI - Differential neurosensory responses of adult Colorado potato beetle, Leptinotarsa decemlineata, to glycoalkaloids. AB - Neurons from chemosensory hairs on the galeae of adult Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), were investigated for responses to glycoalkaloids of the family Solanaceae. While solanine and tomatine elicited irregular firing by multiple neurons and bursting activity at 1 mM concentration in most sensory hairs, stimulation with leptine I resulted in consistently high frequency, slowly adapting responses with a dose-dependent effect between 0.03 and 0.3 mM concentrations. Responses to a mixture of solanine and leptine I suggested possible modification of the leptine I response by other glycoalkaloids, resulting in reduced neural activity relative to leptine I alone. These results establish a method for specifically evaluating leptine I and other glycoalkaloids for effects on feeding behavior of CPB and provide a sensory component for incorporating deterrent chemistry into biorational control methods for the CPB. PMID- 11504018 TI - Electrophysiological and behavioral responses of female Helicoverpa armigera to compounds identified in flowers of African marigold, Tagetes erecta. AB - Seven electrophysiologically active compounds were detected in air-entrained headspace samples of live flowers of Tagetes erecta analyzed by gas chromatography (GC) linked to a female Helicoverpa armigera electroantennograph (EAG) using polar and nonpolar capillary columns. These compounds were subsequently identified using GC linked to mass spectrometry as benzaldehyde, (S) (-)-limonene, (R,S)-(+/-)-linalool, (E)-myroxide, (Z)-beta-ocimene, phenylacetaldehyde, and (R)-(-)-piperitone. Electrophysiological activity was confirmed by EAG with a 1-microg dose of each compound on filter paper eliciting EAG responses that were significantly greater than the solvent control response from female moths. Wind-tunnel bioassays with T. erecta headspace samples, equivalent to 0.4 flower/hr emission from a live flower, elicited a significant increase in the number of upwind approaches from female H. armigera relative to a solvent control. Similarly, a seven-component synthetic blend of EAG-active compounds identified from T. erecta presented in the same ratio (1.0:1.6:0.7:1.4:0.4:5.0:2.7, respectively) and concentration (7.2 microg) as found in the natural sample elicited a significant increase in the number of upwind approaches relative to a solvent control during a 12-min bioassay that was equivalent to that elicited by the natural T. erecta floral volatiles. PMID- 11504019 TI - Close-range attraction in Lygocoris pabulinus (L.). AB - Males of the green capsid bug, Lygocoris pabulinus, exhibit a specific courtship behavior, i.e., a vibration of the abdomen. When both live and dead females were offered to males, this vibration behavior was elicited in most of the males tested. When females were dissected into separate body parts, heads, wings, and legs elicited equal responses, while thorax plus abdomen elicited a much lower response. When separate body parts were extracted, the leg extracts elicited significantly stronger responses than any other extract. This suggests that female L. pabulinus legs are either the source of a close-range sex pheromone or that pheromone is accumulated on the legs due to grooming behavior. The leg extracts contained several hydrocarbons such as n-alkenes, n-alkanes, and some methylalkanes. Female extracts contained more (Z)-9-pentacosene and male extracts contained more (Z)-9-heptacosene. Substrates on which females had walked elicited similar responses as female legs, indicating that the pheromone is deposited on the substrate. This enlarges the functional range of low-volatility compounds, which are thought to function only when sexes are in close vicinity or in contact. PMID- 11504020 TI - Oviposition deterrent by female reproductive gland secretion in Japanese pine sawyer, Monochamus alternatus. AB - Adult females of Monochamus alternatus are known to use palpation to recognize oviposition scars that contain eggs and to be deterred from oviposition. This study investigated the oviposition-deterring activity of a jellylike secretion deposited by the females immediately after oviposition and methanol extracts of female reproductive organs in the laboratory. When females searching for oviposition sites encountered artificial oviposition scars, they stopped walking and drummed the surface and inside of the oviposition scars with their maxillary and labial palpi. When the females encountered the artificial scars plugged with the jellylike secretion, most of them left the scars after palpation. In contrast, when females encountered artificial scars not plugged with the secretion, most of them deposited single eggs through the scars. In another experiment, most females left artificial scars treated with methanol extracts of the spermathecal gland or other reproductive organs after palpation, but most of them oviposited through the scars treated with methanol alone. The results showed that females' recognition of egg-containing scars and departure from such scars were mediated by the chemical(s) produced by their reproductive organs. PMID- 11504021 TI - Reproductive character displacement in Lymantria monacha from northern Japan? AB - Our objective was to test the hypothesis that the pheromone blend and/or diel periodicity of pheromonal communication differ in populations of the nun moth, Lymantria monacha (Lepidoptera: Lymantriidae), from eastern Asia (northern Honshu, Japan) and Central Europe (Bohemia, Czech Republic). Coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of pheromone gland extract of female L. monacha from Japan confirmed the presence of compounds previously identified in pheromone extracts of L. monacha from Bohemia, as follows: (Z)-7-octadecene, 2-methyl-(Z)-7-octadecene (2me-Z7-18Hy), cis-7,8-epoxy octadecane (monachalure), and cis-7,8-epoxy-2-methyloctadecane (disparlure). Field experiments in Honshu suggested that (+)-monachalure is the major pheromone component of L. monacha. 2me-Z7-18Hy significantly enhanced attractiveness of (+) monachalure. Addition of (+)-disparlure to (+)-monachalure plus 2me-Z7-18Hy in Honshu and Bohemia increased attractiveness of lures by 1.2 and 20 times, respectively, indicating that (+)-disparlure is of least and most significance in the respective L. monacha populations. Moreover, capture of male L. monacha in pheromone-baited traps between 18:00 and 24:00 hr in Bohemia and 2:00 and 5:00 hr in Honshu revealed a markedly different diel periodicity of pheromonal communication. Pheromonal communication late at night and use of (+)-monachalure, rather than (+)-disparlure, as the major pheromone component by L. monacha in Honshu may have resulted from interspecific competition with coseasonal L. fumida, which uses the early night for pheromonal communication and (+) disparlure as major pheromone component. Whether communication channel divergence of L. monacha in Honshu indeed constitutes a case of reproductive character displacement is difficult to prove. The evolution of such divergence in sympatric populations of L. fumida and L. monacha would have to be demonstrated. PMID- 11504022 TI - Identification and synthesis of a male-produced sex pheromone from the stink bug Chlorochroa sayi. AB - The reproductive behavior of the stink bug Chlorochroa sayi was studied in the laboratory. There was a sexual maturation period of about 10 days before bugs began mating. Sexually mature adult bugs engaged in courtship consisting of antennation and head-butting of the female by the male, before the female adopted a receptive posture and copulation occurred. Both sexes mated multiple times during their life-spans, with the mean duration of copulations of virgin bugs (42.3 +/- 19.6 min) and experienced bugs (37.3 +/- 28.4 min) being similar. Most matings were initiated in the late afternoon or evening, when pheromone production by males was greatest. Males transferred sperm and nutrients constituting about 17% of their body weight to females during mating. Three male specific components, methyl geranate, methyl citronellate, and methyl (E)-6-2,3 dihydrofarnesoate in a ratio of 100:0.45:1.6, were first detected in volatiles collected from male bugs on green beans about 9-12 days after the final molt to the adult stage. In vertical Y-tube bioassays, females were attracted to odors from mature male bugs, and to a blend of the three male-produced components. Low numbers of females also were attracted in field trials with the three-component blend. The relatively weak attraction may be a result of other, as yet unknown cues being required in addition to the pheromone, such as visual or substrate borne vibrational cues. PMID- 11504023 TI - Components of male aggregation pheromone of strawberry blossom weevil, Anthonomus rubi herbst. (Coleoptera:Curculionidae). AB - The strawberry blossom weevil, Anthonomus rubi, is a major pest of strawberries in the United Kingdom and continental Europe. As part of a project to develop noninsecticidal control methods, the pheromone system of this species was investigated. Comparison of volatiles produced by field-collected, overwintering individuals of each sex led to identification of three male-specific compounds- (Z)-2-(3,3-dimethylcyclohexylidene)ethanol, (cis)-1-methyl-2-(1 methylethenyl)cyclobutaneethanol, and 2-(1-methylethenyl)-5-methyl-4-hexen-1-ol (lavandulol)--in amounts of 6.1, 1.2, and 0.82 microg/day/ male. The first two compounds are components of the aggregation pheromone of the boll weevil, Anthonomus grandis, grandlure II and grandlure I, respectively. Grandlure I was the (1R,2S)-(+) enantiomer and lavandulol was a single enantiomer, although the absolute configuration was not determined. Trace amounts of the other two grandlure components (Z)-(3,3-dimethylcyclohexylidene)acetaldehyde (grandlure III) and (E)-(3,3-dimethylcyclohexylidene)acetaldehyde (grandlure IV) were also detected. (E,E)-1-(1-Methylethyl)-4-methylene-8-methyl-2,7-cyclo-decadiene (germacrene-D), a known volatile from strawberry plants, Fragaria ananassa, was collected in increased amounts in the presence of pheromone-producing weevils. Male weevils only produced pheromone on F. ananassa and not on scented mayweed, Matracaria recutita, or cow parsley, Anthriscus sylvestris, although these are known food sources. In field trials using various combinations of synthetic grandlures I, II, III, and IV and lavandulol, significantly more weevils were caught in traps baited with blends containing grandlure I and II and lavandulol than in those baited with blends without lavandulol or unbaited controls. Addition of grandlure III and IV had no significant effect on attractiveness. Horizontal sticky traps were found to be more effective than vertical sticky traps or standard boll weevil traps. In mid-season females predominated in the catches, but later more males than females were trapped. PMID- 11504024 TI - Novel chemistry of abdominal defensive glands of nymphalid butterfly Agraulis vanillae. AB - Abdominal defensive glands of both sexes of the Gulf fritillary butterfly, Agraulis vanillae (Linnaeus) (Nymphalidae:Heliconiinae) emit a pronounced odor when disturbed. We have identified 6-methyl-5-hepten-2-one; oleic, palmitic, and stearic esters of the corresponding alcohol 6-methyl-5-hepten-2-ol; hexadecyl acetate; 1,16-hexadecanediol diacetate; and 1,15-hexade-canediol diacetate in the glandular exudate. Since we have determined that free-flying birds or birds in a butterfly conservatory discriminate against A. vanillae as prey, we suggest that the constituents in the glands may play a defensive role against potential avian predators. PMID- 11504025 TI - Lack of intraspecific aggression in the ant Tetramorium bicarinatum: a chemical hypothesis. AB - Tetramorium bicarinatum (Myrmicinae) is an ant species frequently found in tropical and subtropical areas, particularly in Africa, Southeast Asia (Japan), and South America (Brazil). The species is polygynous, reproduces by budding, and has sterile workers. Since the nests are widely distributed in a given area, the problem arises of territorial defense against conspecifics. Because not all ants defend territories, we assessed the defensive behavior of T. bicarinatum workers through intraspecific and interspecific aggressiveness tests. A detailed behavioral study of the interactions between workers from several different colonies of T. bicarinatum (originating from Japan and Brazil) showed that workers do not discriminate against conspecific nonnestmate individuals, but they are highly aggressive towards allospecifics (Myrmica rubra, Myrmicinae). The results suggest that each colony from this ant species possesses a similar colonial odor. Chemical analyses of the cuticular hydrocarbons of these species were made with gas chromatography coupled to mass spectrometry. Results showed that the different colonies of T. bicarinatum possess a common chemical profile mainly composed of straight-chain alkanes and alkenes, while M. rubra possess more methyl-branched alkanes. We suggest that methyl alkane cues play a determining role in colonial recognition and that these results could explain the underlaying basis of the lack of intraspecific aggressiveness in T. bicarinatum. PMID- 11504026 TI - Avoidance of conspecific injury-released chemical cues by free-ranging Gammarus lacustris (Crustacea:Amphipoda). AB - Behavioral responses to chemical cues have been demonstrated for a range of aquatic animals. Injury-released chemical alarm cues from conspecifics are released when a prey's predator is actively foraging. Detection of these cues elicits antipredator behaviors that reduce the probability of predation. Amphipod crustaceans in the genus Gammarus are widespread denizens of ponds and streams. Antipredator responses by Gammarus to conspecific alarm cues, and subsequent reduction of predation risk, are known from experiments in the laboratory. Here, we verify laboratory findings by demonstrating an avoidance response to alarm cues using a field population of G. lacustris. We used small traps baited with sponge blocks containing either water (control) or injury-released cues from Gammarus. We repeated the experiment twice. In both experiments, significantly fewer Gammarus were captured in traps with alarm cue sponges than in traps with water sponges. Predatory leeches Dina parva were attracted to Gammarus traps in the first experiment but not the second experiment. In the second experiment, we measured the individual weight of captured amphipods. Two size classes were present; small (1-5 mg) and large (35-108 mg). Both sizes contributed to the avoidance response. Within the large size class, small individuals were significantly less responsive to the alarm cue than large individuals, implying that small adult Gammarus may have different cost/benefit decision criteria for risk assessment than large Gammarus. PMID- 11504027 TI - Mammalian exocrine secretions XV. Constituents of secretion of ventral gland of male dwarf hamster, Phodopus sungorus sungorus. AB - In a study aimed at the chemical characterization of constituents of the ventral gland secretion of the male dwarf hamster, Phodopus sungorus sungorus, 48 compounds, including saturated alcohols, saturated and unsaturated ketones, saturated and unsaturated straight-chain carboxylic acids, iso- and anteisocarboxylic acids, 3-phenylpropanoic acid, hydroxyesters, 2-piperidone, and some steroids were identified in the secretion. The position of the double bonds in gamma-icosadienyl-gamma-butyrolactone and gamma-henicosadienyl-gamma butyrolactone, and the position of methylbranching in seven C16-C21 saturated ketones could not be established. Several constituents with typically steroidal mass spectra also remained unidentified. The female dwarf hamster's ventral gland either does not produce secretion or produced so little secretion that it was impossible to collect enough material for analysis. PMID- 11504028 TI - Mammalian exocrine secretions XVI. Constitutents of secretion of supplementary sacculi of dwarf hamster, Phodopus sungorus sungorus. AB - As a first step in a study of the role of the secretion of the supplementary sacculi (buccal secretion) of the dwarf hamster, Phodopus sungorus sungorus, almost complete chemical characterization of the secretion was achieved. The 35 compounds identified include carbon dioxide, hydrogen sulfide, a large number of carboxylic acids (representing the bulk of the organic volatile fraction of the secretion), phenol, 2-piperidone, indole, two long-chain hydroxyesters, cholestrol, desmosterol, and lanosterol. The position of the double bonds in gamma-icosadienyl-gamma-butyrolactone and gamma-henicosadienyl-gamma butyrolactone could not be determined, and these two compounds remained only partially characterized. Large variations were found in the relative concentrations in which the short-chain carboxylic acids are present in the secretions of individual animals, and although this aspect was not investigated in sufficient detail in the present investigation, the difference in the carboxylic acid profiles of the secretions of individual animals could play a role in individual recognition in this animal. PMID- 11504029 TI - Effects of genotype, nutrient availability, and defoliation on aspen phytochemistry and insect performance. AB - Genetic and environmental variability, and their interactions, influence phytochemical composition and, in turn, herbivore performance. We evaluated the independent and interactive effects of plant genotype, nutrient availability, and defoliation on the foliar chemistry of quaking aspen (Populus tremuloides) and consequences for performance of gypsy moths (Lymantria dispar). Saplings of four genotypes were grown under two conditions of nutrient availability and subjected to three levels of artificial defoliation. Concentrations of all secondary and primary metabolites evaluated responded to at least one or more of the experimental treatments. Of the secondary metabolites, phenolic glycosides were affected strongly by genotype, less so by nutrient availability, and not induced by defoliation. Condensed tannins were strongly dependent upon genotype, soil nutrient availability, and their interaction, and, in contrast to phenolic glycosides, were induced by artificial defoliation. Of the primary metabolites, foliar nitrogen was affected by genotype and soil nutrient availability. Starch concentrations were affected by genotype, nutrient availability, defoliation and interactions among these factors. Foliar water content responded to genotype, nutrient availability, and defoliation, and the effect of nutrient availability depended on genotype. Herbivore performance on these plants was strongly influenced by plant genotype and soil nutrient availability, but much less so by defoliation. Although several of the compound types (condensed tannins, starch, and water) responded to defoliation, quantitative variation in these compounds did not contribute to substantive changes in herbivore performance. Rather, the primary source of variation in insect performance was due to plant genotype (phenolic glycoside levels), while nutrient availability (foliar nitrogen levels) was of secondary importance. These results suggest that genetic variation in aspen plays a major role in determining patterns of insect performance, whereas environmental variation, such as was tested, here is of negligible importance. PMID- 11504030 TI - Importance of bacterial decomposition and carrion substrate to foraging brown treesnakes. AB - Brown treesnakes are an invasive species to the island of Guam that have caused extensive ecological and economic damage. Efforts to control the snake population have included trapping using live mouse lures, but for logistical and economic reasons a synthetic lure is needed. When searching for live food, brown treesnakes use both visual and odor cues. However, when searching for carrion, odor cues are sufficient. Attempts to develop synthetic lures based on chemical reconstruction of the complex carrion odor have not succeeded. We provide evidence that a microbial-substrate interaction is important for bait take by brown treesnakes. Microbial cultures taken from mouse carrion indicate that Enterobacter agglomerans is the predominant bacterium, and field tests suggest that this organism may be important to odor production that attracts brown treesnakes. This information may prove useful in the development of microbial based biological reactors that could be formulated to produce a continuous stream of odor of sufficient complexity so as to be attractive to foraging snakes. PMID- 11504031 TI - Effects of predator chemical cues and behavioral biorhythms on foraging activity of terrestrial salamanders. AB - Red-backed salamanders, Plethodon cinereus, show a variety of alarm responses to chemical cues from eastern garter snakes, Thamnophis sirtalis. We measured the foraging activity of red-backed salamanders exposed to water soiled by a garter snake (fed P. cinereus) or to unsoiled water. Salamanders exposed to snake-soiled water showed less foraging activity than salamanders exposed to unsoiled water; therefore, predators could have nonlethal effects on salamander populations. Our results also show additional factors influenced salamander foraging activity. Salamander foraging activity and responsiveness to chemical cues do not appear to have been affected by sex or food deprivation. Salamander foraging activity does appear to have been influenced by activity biorhythms. Foraging activity of animals in both treatments showed a bimodal periodicity that is consistent with natural activity patterns controlled by internal biorhythms. Exposure to snake soiled water significantly reduced foraging activity during periods of peak foraging activity, but had a subtler effect on foraging activity during natural lulls in activity. We suggest that both activity biorhythms and exposure to chemical cues are important factors affecting salamander foraging behavior. PMID- 11504032 TI - Sequestration of host plant-derived compounds by geometrid moth, Milionia basalis, toxic to a predatory stink bug, Eocanthecona furcellata. AB - A predatory stink bug, Eocanthecona furcellata, died after feeding on Milionia basalis larvae. The compounds toxic to E. furcellata were isolated from the hemolymph of M. basalis larvae and identified as inumakilactone A, nagilactone C, and nagilactone C glucoside. The concentrations of inumakilactone A, nagilactone C, and nagilactone C glucoside in the hemolymph of the final instar larvae were 130, 50, and 770 microg/ml, respectively. Nagilactone C showed the highest insecticidal activity against second-instar nymphs of E. furcellata, while nagilactone C glucoside showed the lowest, one twentieth of that of nagilactone C. When mixed compounds were given at the same concentrations as those in hemolymph of M. basalis, all nymphs of E. furcellata died with in three days. Inumakilactone A and nagilactone C were found to be in the leaves of podocarp, Podocarpus macrophyllus, the only host plant of M. basalis, at concentrations of 13 and 175 microg/g fresh weight, respectively. However, no nagilactone C glucoside was detected in the leaves of this species. These results suggested that M. basalis may transform nagilactone C to its glucoside. PMID- 11504033 TI - Comparison of cultivars of ornamental crop Gerbera jamesonii on production of spider mite-induced volatiles, and their attractiveness to the predator Phytoseiulus persimilis. AB - We investigated whether volatiles produced by spider mite-damaged plants of four gerbera cultivars differ in attractiveness to Phytoseiulus persimilis, a specialist predator of spider mites, and how the mite-induced odor blends differ in chemical composition. The gerbera cultivars differed in resistance, as expressed in terms of spider mite intrinsic rate of population increase (rm). In order of increasing resistance these were Sirtaki, Rondena, Fame, and Bianca. To correct for differences in damage inflicted on the cultivars, we developed a method to compare the attractiveness of the blends, based on the assumption that a larger amount of spider mite damage leads to higher attraction of P persimilis. Spider mite-induced volatiles of cultivars Rondena and Bianca were preferred over those of cultivar Sirtaki. Spider mite-induced volatiles of cultivars Sirtaki and Fame did not differ in attractiveness to P. persimilis. Sirtaki plants had a lower relative production of terpenes than the other three cultivars. This was attributed to a low production of cis-alpha-bergamotene, trans-alpha-bergamotene, trans-beta-bergamotene, and (E)-beta-farnesene. The emission of (E)-beta-ocimene and linalool was lower in Sirtaki and Fame leaves than in Bianca and Rondena. The importance of these chemical differences in the differential attraction of predatory mites is discussed. PMID- 11504034 TI - Single and blended maize volatiles as attractants for diabroticite corn rootworm beetles. AB - Synthetic maize volatiles and analogs dispensed singly and blended were tested for attractiveness to western (WCR, Diabrotica virgifera virgifera) and northern corn rootworm beetles (NCR, D. barberi) in maize fields. Newly identified attractants included syn-benzaldoxime, especially for NCR, and beta-caryophyllene for WCR females. (+/-)-Linalool was more effective than was (-)-linalool. Myrcene, (+)-beta-pinene, and (-)-beta-pinene were unattractive. Adding methyl salicylate to (+/-)-linalool, (+)-alpha-terpineol, or beta-ionone appeared to synergistically increase capture of WCR females, but dispensing the terpenes in binary blends did not. Dose-response data for methyl salicylate, (+/-)-linalool, and a blend of both compounds confirmed the synergy. beta-Caryophyllene, but not (-)-alpha-pinene, added to the latter blend produced a further synergistic increase in WCR female capture that did not vary with sesquiterpene dose from 1.0 to 100 mg. Indole addition to the same blend caused an increase in WCR female captures indicative of synergy, assuming that each did not individually lure different segments of the WCR female population. The green leaf volatiles (Z)-3 hexenyl acetate and (Z)-3-hexen-1-ol were unattractive alone and had no influence on efficacy of traps baited with 3.3 mg each of (+/-)-linalool, methyl salicylate, and beta-caryophyllene. The latter mixture captured about half as many WCR females as did 10 mg of 4-methoxycinnamaldehyde, a potent WCR attractant standard. Substituting beta-ionone for (+/-)-linalool yielded a ternary blend that captured more beetles than did the aldehyde and was unaffected by aldehyde addition. Olive oil, which has been used to sustain attractant volatilization, did not affect captures. The results show that the blending of maize volatiles has thepotential to greatly improve efficacy of lures having promising applications in corn rootworm population management. PMID- 11504035 TI - A chiral sex pheromone system in the pea midge, Contarinia pisi. AB - The sex pheromone of the pea midge consists of 2-acetoxytridecane, (2S,11S) diacetoxytridecane and (2S,12S)-diacetoxytridecane. The responses of male pea midges to the corresponding stereoisomers of (2S,11S)-diacetoxytridecane and (2S,12S)-diacetoxytridecane were tested in field trapping experiments and by electroantennographic recordings. When added at 20% of the pheromone component to the sex pheromone blend, the (2S,11R)- and (2R,11S)-stereoisomers of (2S,11S) diacetoxytridecane, were shown to have a strong inhibitory effect on male attraction in the field. At the same dose, (2R,11R)-diacetoxytridecane, (2R,12R) diacetoxytridecane, and meso-2,12-diacetoxytridecane, did not have a significant effect on male behavior. It was also shown that substitution of either (2S,11S) diacetoxytridecane or (2S,12S)-diacetoxytridecane with the related stereoisomers reduced trap catches to the level of blank traps. The electroantennographic recordings showed similar dose-response curves for the pheromone components and the stereoisomers shown to have an inhibitory effect. It seems likely that male antennae have receptors for bothpheromone components and for inhibitory stereoisomers. Scanning electron microscopy and transmission electron microscopy of the antennae revealed three types of sensilla involved in chemoreception: sensilla circumfila, sensilla trichodea, and sensilla coeloconica. The sensilla circumfila and trichodea are both innervated by two sensory cells, whereas the sensilla coeloconica are innervated by four to five cells. PMID- 11504036 TI - Sex pheromone components of the buck moth Hemileuca maia. AB - The sex attractant pheromone blend of Hemileuca maia (Lepidoptera: Saturniidae) from the vicinity of Baton Rouge, Louisiana, has been identified. The major component of the blend is (E10,Z12)-hexadeca-10,12-dienal (E10,Z12-16:Ald), in combination with the minor components (E10,Z12)-hexadeca-10,12-dien-1-ol (E10,Z 12-16:OH), and (E10,Z12)-hexadeca-10,12dien-1-yl acetate (E10,Z12-16:Ac). Ratios of the compounds in extracts of female pheromone glands varied around a mean of 100:7.4:6.3. None of the three components were attractive to male moths when tested as single components. Several other compounds were tentatively identified from female pheromone gland extracts, including E10,E12-16:Ald, E10,E12-16:OH, and E10,E12-16:Ac, but addition of these components, either alone or in combination, at biologically relevant rates, did not significantly increase the attractiveness of lures. The saturated analogs, hexadecanal, hexadecanol, and hexadecyl acetate, also were identified in gland extracts, but had no apparent effect as pheromone components. PMID- 11504037 TI - The nature of the gregarizing signal responsible for maternal transfer of phase to the offspring in the desert locust Schistocerca gregaria. AB - We examined aggregative behavior of hatchlings of the desert locust Schistocerca gregaria emerging from solitaria egg pods that had been incubated in sand previously used for consecutive ovipositions by gregarious females. Hatchlings derived from such eggs were significantly more gregarious than those derived from pods laid in clean sand. The gregarizing effect of the sand-associated factor originating from 3, 5, or 10 ovipositions by gregarious females increased in a dose-dependent fashion. Washing the sand with organic solvents following such ovipositions, or flushing it with nitrogen gas, led to substantial loss of its gregarizing effect, showing that the active signal is volatile and of medium polarity. The gregarizing activity of the exposed sand correlated with the presence of C-8 unsaturated ketones, (Z)-6-octen-3-one and (E,E)-3,5-octadiene-2 one and its E,Z isomer, previously shown by us to form part of the releaser pheromone system that mediates group oviposition in S. gregaria. These ketones were present in relatively large amounts in the eggs obtained from egg pods of gregarious females and were also detected in the extracts of accessory glands of gregarious females, a candidate source of the gregarization factor. It is proposed that the pheromone is secreted at the onset of oviposition. This would account for its distribution predominantly within the eggs and surrounding sand at the site of oviposition. The study sheds new light on the pheromonal mechanism associated with transgenerational transmission of gregarious characters in crowded S. gregaria populations and provides yet another case in this insect of dual releaser and primer roles played by the same pheromone blend. PMID- 11504038 TI - (Z)-3-hexenyl (R)-3-hydroxybutanoate: a male specific compound in three North American decorator wasps Eucerceris rubripes, E. conata and E. tricolor. AB - The males of the decorator wasps Eucerceris have been observed to display abdomen dragging behavior on plants surrounding their nest. It is thought that this applies a territorial-marking sex pheromone that serves to alert females to the males' territory for courtship and mating. The extracts of three species E. rubripes, E. conata, and E. tricolor have been analyzed by GC-MS. The gas chromatograms revealed the presence, in large quantities, of one common volatile compound in the male head extracts. By analogy with the closely related bee wolves Philanthus, we believe the pheromone is produced in the mandibular glands. Utilizing mass spectrometry, microreactions, FT-IR, and 1H and 13C NMR spectroscopy the structure of the pheromone was proposed to be (Z)-3-hexenyl 3 hydroxybutanoate, which was confirmed by synthesis. The absolute configuration of the chiral center was determined to be R for the three species by preparing the Mosher esters of the insect samples and comparing their GC retention times with a synthetic sample of known absolute configuration. In addition 2- and 3-hexenoic acid and some aromatic compounds were also found in varying quantities in both males and females along with hydrocarbons and fatty acids, although no species specific profiles emerged. PMID- 11504039 TI - Correspondence of soldier defense secretion mixtures with cuticular hydrocarbon phenotypes for chemotaxonomy of the termite genus Reticulitermes in North America. AB - Soldier defense secretions from samples of Reticulitermes collected in California, Nevada, Arizona, New Mexico, and Georgia were characterized and correlated with cuticular hydrocarbon phenotypes. Twenty-seven cuticular hydrocarbon phenotypes have been defined, and soldier defense secretion (SDS) phenotypes have been described for 25 of these. Forty-five terpenoid compounds were found, including monoterpenes, sesquiterpenes, and a few diterpenes. The monoterpenes include (-)-alpha-pinene, (-)-beta-pinene, (-)-camphene, myrcene, (Z)- and (E)-ocimene, and (-)-limonene. The major sesquiterpenes produced are (+) gamma-cadinene, (+)-gamma-cadinene aldehyde, (-)-germacrene A, germacrene B, gamma-himachalene, and beta-bisabolene. Some SDS phenotypes pair with more than one cuticular hydrocarbon phenotype; however, with two exceptions, each hydrocarbon phenotype is associated with only one SDS phenotype. These chemical characterizations lend support to the conclusion that there are numerous undescribed species of Reticulitermes in North America. PMID- 11504040 TI - A model for peak and width of signaling windows: Ips duplicatus and Chilo partellus pheromone component proportions--does response have a wider window than production? AB - Pheromone communication systems have a reliable signal with a restricted window of amounts and ratios released and perceived. We propose a model based on a Gaussian response profile that allows a quantification of the response peak (location of optimum) and a measure of the peak width (response window). Interpreting the Gaussian curve, fitted by nonlinear regression (NLR), as a standard normal distribution, the peak location equals the mean (it) and the window width equals 2 x the standard deviation (2sigma). The NLR procedure can provide an objective measure for both peak location and width for a wide range of data sets. Four empirical data sets as well as 10 literature data sets were analyzed. The double-spined spruce engraver, Ips duplicatus, was field tested in four populations to find the optimum proportion for attraction to the two male aggregation pheromone components, ipsdienol (Id) and (E)-myrcenol(EM), ranging from 0 to 100% of Id. Tests in Norway and the Czech Republic confirmed the preference of western populations for a blend between 50 and 90% Id. A population in Inner Mongolia showed a preference for traps with the 10 and 50% Id baits. The NLR fitted values for response peak and width (mu; 2sigma) were: Norway 0.64, 0.73; Czech Republic 0.53, 0.73; NE China 0.77, 0.29; and Inner Mongolia 0.33, 0.50. The signal produced by Norwegian field-collected males had a narrower window width (2sigma = 0.12). Males of the maize stem borer, Chilo partellus, were tested in a flight tunnel for their response to variation in the two major female sex pheromone gland components, (Z)- l1-hexadecenal and the corresponding alcohol (OH). Variation of the alcohol in seven levels from 2 to 29% OH showed the highest male response for 17% OH. For all behavioral steps, the peak of male response was near mu = 0.14, while the window width fell from 2sigma = 0.5 to 0.2 for eight sequential behavioral steps from take-off to copulation. Female production had a similar peak location (mu = 0.13) but a narrower width, 2sigma = 0.14. Literature data from other moth species showed similar patterns, with a wider male response relative to the female production windows. Literature data on response to enantiomer ratios in a hymenopteran and to pheromone amounts in a dipteran were also described by our model. In a bark beetle population (Ips pini), with two hybridizing enantiomeric strains, the production peaks were narrower (0.1) than the response peaks (0.5). Thus, it in general, seems that in the pheromone systems analyzed, the width of the response window (2sigma = 0.1 to 0.8) is larger than that of the production window (2sigma = 0.03 to 0.14), irrespective of the sex of the sender. PMID- 11504041 TI - Density-dependent chemical interference--an extension of the biological response model. AB - The response of plants to many phytochemicals changes from stimulatory to inhibitory as the concentration of the phytochemical increases. In this paper, a previous biological response model is extended to yield estimates of plant responses to changes in phytochemical concentrations in the case of density dependent phytotoxicity. This requires a knowledge of plant densities, phytochemical concentrations in soil, and the relationship between the two. According to this model extension, inhibition is a probable outcome in density dependent chemical interference, but phytotoxic effects may become stimulatory as plant density increases. In addition, low phytochemical concentrations in soil may cause an increase in the slope of the biomass-density relationship compared to the slope of control treatments. Experimental data from the literature support this model extension, and in several cases density-dependent chemical interference can be estimated mathematically. PMID- 11504042 TI - Association of carbon disulfide with plants in the family Fabaceae. AB - We examined 40 taxa from nine genera within the subfamily Mimosoideae; 29 (73%) from six genera produced carbon disulfide (CS2). In addition, 19 of 40 taxa (48%) produced carbonyl sulfide (OCS). Of nine mimosoid taxa that produced CS2, all possessed a djenkolic acid and a cysteine lyase. Of three mimosoid taxa that did not produce CS2, two lacked a cysteine lyase and one lacked both a lyase and a djenkolic acid. Of 16 taxa from 14 genera from the other two subfamilies of the Fabaceae, the Caesalpinioideae and Papilionoideae, none produced CS2. The results suggest that CS2 production is common in the Mimosoideae and uncommon in the Caesalpinioideae and Papilionoideae and that plants in the Mimosoideae that do produce CS2 must possess both a djenkolic acid and a cysteine lyase for this production to occur. PMID- 11504043 TI - Chiral analysis of UV nonabsorbing compounds by capillary electrophoresis using macrocyclic antibiotics: 1. Separation of aspartic and glutamic acid enantiomers. AB - Glycopeptide antibiotics, namely vancomycin or teicoplanin, were evaluated in capillary electrophoresis for the analysis of UV nonabsorbing compounds such as aspartic and glutamic acid enantiomers. Electrophoretic runs were performed in laboratory-made polyacrylamide-coated capillaries using the partial filling counter current method in order to avoid the presence on the detector path of the absorbing chiral selector. The background electrolyte consisted of an aqueous or aqueous-organic buffer in the pH range of 4.5-6.5 of sorbic acid/histidine and the appropriate concentration of chiral selector. Several experimental parameters such as antibiotic concentration and type, buffer pH, organic modifier, type and concentration of absorbing co-ion (for the indirect UV detection) were studied in order to find the optimum conditions for the chiral resolution of the two underivatized amino acids in their enantiomers. Among the two investigated chiral selectors, vancomycin resulted to be the most useful chiral selector allowing relatively high chiral resolution of the studied compounds even at low concentration. The optimized method (10 mM sorbic acid/histidine, pH 5, and 10 mM of vancomycin) was used for the analysis of real samples such as teeth dentine and beer. PMID- 11504044 TI - Interaction between 18-crown-6-tetracarboxylic acid and positional substituents of enantiomers and simultaneous separation of positional enantiomers of methyl-DL tryptophans by capillary electrophoresis. AB - 18-Crown-6-tetracarboxylic acid (18C6H4) is a chiral selector with high selectivity for the enantioseparation of solutes bearing the primary amine function. This work presents the simultaneous separation of positional enantiomers of methyl-DL-tryptophans by using 18C6H4 as an additive to the background electrolyte. Separation conditions such as pH, the concentration of 18C6H4, and the applied voltages have critical inference on the simultaneous separation. The addition of cyclodextrins as anionic surfactants to the background electrolyte did not improve the separation. The selector-selectand interactions between 18C6H4 and the positional enantiomers have been investigated. It was observed that both the position and type of substituents contribute to the enantioselectivity. The migration order and resolution depended on the distance from the substituents to the asymmetric carbon of the enantiomers. PMID- 11504045 TI - Salting-out solvent extraction for the off-line preconcentration of benzalkonium chloride in capillary electrophoresis. AB - The use of salting-out effect for the off-line preconcentration of charged analytes in capillary electrophoresis is demonstrated for the first time. Using benzalkonium chloride (BAK) as model compound, a mixture of cationic surfactants consisting of even-numbered alkylbenzyl quaternary ammonium homologues (C8-C18), the addition of appropriate amounts of sodium chloride and acetonitrile in the sample solution (2 mL sample volume) was found to be capable of providing ca. 40 fold enhancement in detection sensitivity. In addition to affording a preconcentrating effect due to the extraction of BAK in the smaller volume water miscible organic solvent phase (acetonitrile), the organic solvent also serves to improve the peak area and shape of the longer chain surfactants. Optimal experimental conditions, such as volume of acetonitrile and concentration of sodium chloride, for the preconcentration of BAK with good preconcentration factors and reproducibility were investigated. The usefulness of the present method was demonstrated for the improved determination of BAK present in commercially available industrial and pharmaceutical products. PMID- 11504046 TI - Electrophoretic migration behavior of DNA fragments in polymer solution. AB - A newly developed polymer coil shrinking theory is described and compared with the existing entangled solution theory to explain electrophoretic migration behaviour of DNA in hydroxypropylmethylcellulose (HPMC) polymer solution in buffer containing 100 mM tris(hydroxymethyl)aminomethane 100 mM boric acid, 2 mM ethylenediaminetetraacetic acid at pH 8.3. The polymer coil shrinking theory gave a better model to explain the results obtained. The polymer coil shrinking concentration, Cs, was found to be 0.305% and the uniform entangled concentration, C+, 0.806%. The existence of three regions (the dilute, semidilute, and concentrated solution) at different polymer concentrations enables a better understanding of the system to guide the selection of the best conditions to separate DNA fragments. For separating large fragments (700/ 800 bp), dilute solutions (HPMC < 0.3%) should be used to achieve a short migration time (10 min). For small fragments (200/300 bp), concentrated solutions are preferred to obtain constant resolution and uniform separation. The best resolution is 0.6% HPMC due to a combined interaction of the polymer coils and the entangled structure. The possibility of DNA separation in semidilute solution is often neglected and the present results indicate that this region has a promising potential for analytical separation of DNA fragments. PMID- 11504047 TI - Stacking and separation of coproporphyrin isomers by acetonitrile-salt mixtures in micellar electrokinetic chromatography. AB - The effectiveness of the addition of salt and acetonitrile in the sample matrix to induce narrowing of the analyte zones is demonstrated for the first time in micellar electrokinetic chromatography (MEKC). Using coproporphyrin (CP) I and III isomers as test compounds, the use of sodium cholate (SC) as the micelle in the separation buffer and a high concentration of sodium chloride in the aqueous sample solution (without the presence of an organic solvent) were found to provide enhancement in peak heights for both CP I and III, but yielded very poor resolution of these two positional isomers at sample size of 10% capillary volume or larger. With the addition of acetonitrile as the organic solvent in the aqueous sample solution (acetonitrile-salt mixtures), baseline/partial resolution of CP I and III was obtained even at large injection volumes, along with significant increase in peak heights for both isomers. Possible mechanisms responsible for the narrowing of analyte zones are briefly discussed. The effects of experimental parameters, such as concentrations of salt and acetonitrile, on peak heights and resolution of the test compounds were studied. Importantly, the usefulness of the present method was demonstrated for the MEKC determination of endogenous CP I and III present in normal urine samples with good separation and detection performances. PMID- 11504048 TI - Sol-gel technique for the preparation of beta-cyclodextrin derivative stationary phase in open-tubular capillary electrochromatography. AB - A novel open-tubular capillary electrochromatography (OT-CEC) column coated with 2,6-dibutyl-beta-cyclodextrin (DB-beta-CD) was prepared using sol-gel technique. In the sol-gel approach, owing to the three-dimensional network of sol-gel and the strong chemical bond between the stationary phase and the surface of capillary columns, good chromatographic characteristics and unique selectivity in separating isomers were shown. We achieved high efficiencies of 5-14 x 10(4) plates/m for the isomeric nitrophenols using the sol-gel-derived DB-beta-CD columns. The migration time reproducibility of the separation of the isomeric nitrophenols was better than 2.2% over five runs and 4.5% from column to column. These sol-gel-coated DB-beta-CD columns have shown improved separations of isomeric aminophenols, isomeric dihydroxybenzenes and isomeric nitrophenols, in comparison with the sol-gel matrix capillary column. The influences of buffer pH and methanol solvent on separation were investigated. The chiral resolution of enantiomers such as ibuprofen and binaphthol was explored primarily. PMID- 11504049 TI - Influence of buffer composition and sample pretreatment on efficiency separation for monitoring neuropeptides in plasma using capillary electrophoresis. AB - More efficient and faster separation conditions for qualitative as well as quantitative analysis of neuropeptides in human plasma using capillary zone electrophoresis (CZE) have been developed. The analysis method for neuropeptides has been improved specifically to study thyroid hormone related neuropetides for the regulation of thyroid disease. In this study, we investigated the pretreatment methods, composition of the running buffer and rinsing procedures between runs in order to obtain more sensitive and faster separation of trace neuropeptides in plasma by CZE. The tested neuropeptides were somatostatin (SOMA), vasopressin (VP), neurotensin (NT), and thyrotropin-releasing hormone (TRH). Plasma samples were pretreated by deproteinization and solid-phase extraction method. The fraction of neuropeptides was reconstituted in 40% acetonitrile followed by ultrafiltration, and then analyzed by CZE. Resolution and sensitivity was improved using the separation buffer composition with 100 mM Tris-phosphate buffer (pH 2.0) while the sensitivity was further improved via a stacking method using the sample buffer of 40% acetonitrile. These sample pretreatment methods and buffer condition permit quantitative analysis on tested neuropeptides at the 20 ng/mL level. The rinsing procedures between runs using 90% ethanol dramatically shortened the rinsing time to 30 min. PMID- 11504050 TI - Separation of chlorophenoxyacetic acids and chlorophenols by using capillary zone electrophoresis. AB - In this study, the choice of electrolyte systems for the separation and detection of a range of chlorophenoxyacetic acids and chlorophenols by means of capillary zone electrophoresis (CZE) is discussed. A series of acetate buffers over the buffering capacity pH range 4.03-5.5 were initially chosen for the separation. It was found that chlorophenoxyacetic acids could be separated at pH 4.03 and 4.5 but the most satisfactory separation of chlorophenols was obtained at pH 5.5. The factors affecting separation selectivity, including the addition of organic modifiers, was also studied. The use of 25% 2-butanol, 5% ethylene glycol and 10% acetonitrile as organic solvents resulted in the total separation of both classes of these compounds but poor peak shape of chlorophenols resulted and a number of chlorophenoxyacetic acids were not well separated. A borate-phosphate buffer gave improved peak shape of chlorophenols. Further improved separation of the components of the mixture was obtained by the addition of 2 mM fully methylated beta-cyclodextrin to the 35 mM borate- 60 mM phosphate buffer at pH 6.5, maintaining good peak shape. In this case, separation of the two compound classes, chlorophenoxyacetic acids and chlorophenols, is achieved, with complete resolution of individual compounds in less than 5 min with high efficiency (of the order of 150,000 plates for the ca. 40 cm column). The method is applied to a commercial 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide mixture. PMID- 11504051 TI - Metal speciation using capillary electrophoresis--inductively coupled plasma atomic emission spectrometry and polytetrafluoroethylene capillaries. AB - A capillary electrophoresis--inductively coupled plasma atomic-emission spectrometry (CE-ICP-AES) system using a polytetrafluoroethylene (PTFE) capillary has been developed. The CE-ICP interface was a modified concentric nebulizer. The PTFE capillary (50 microm internal diameter) was used as the central capillary of the nebulizer. Using the PTFE capillaries, the solution flow rate induced by the carrier gas flow was smaller than that of glass capillary. Solution flow was mainly induced by the CE electric field. Baseline separation of Ba2+/Mg2+ ion pair using simple buffer solution of 0.014 M sodium acetate was reported. Separation and correlation of metal species in metallothioneins (MT-1 and MT-2 in MT) of rabbit liver using the CE-ICP system were also discussed. PMID- 11504052 TI - Determination of trace metals by capillary electrophoresis. AB - A new analytical procedure is developed using a strong complexing agent, 1,10 phenanthroline (Phen), for direct UV detection of Zn, Mn, Cu, Co, Cd, and Fe at microg/L concentrations in environmental water samples. The metal chelates formed showed different electrophoretic mobilities and solved the comigration problem for capillary electrophoresis (CE) separation of free metal ions. To obtain stable metal-Phen chelates during the capillary zone electrophoresis (CZE) run, both pre-column and on-column complexation are required and threefold excess of Phen over metal ions should be added to the sample. The optimized background electrolyte (BGE) consists of 30 mM hydroxylamine hydrochloride and 0.1% methanol at pH 3.6. Under hydrodynamic sampling, CE run at + 20 kV in 65 cm x 0.05 mm ID fused-silica column with detection at 265 nm, baseline separation, satisfactory working ranges (10 microg/L to 5.5 mg/L), sensitive detection limits (1-3 microg/L), good repeatability for migration times (relative standard deviation, RSD 0.36-0.81%, n = 5), peak area (RSD 3.2-4.2%, n = 5) and peak height (RSD 3.2 4.5%, n = 5) were obtained for the metal cations investigated. The reliability of the method was established by parallel determination using the inductively coupled plasma-atomic emission spectrometry (ICP-AES) method giving results within statistical variation. The procedure developed is shown to provide a quick, sensitive, precise, and economic method for simultaneous determination of metal cations that can form stable chelates with Phen. PMID- 11504053 TI - Analyses of quaternary ammonium drugs in horse urine by capillary electrophoresis mass spectrometry. AB - A capillary electrophoresis-mass spectrometry (CE-MS) method for the analysis of quaternary ammonium drugs in equine urine was developed. Quaternary ammonium drugs were first extracted from equine urine by ion-pair extraction and then analysed by CE-MS in the positive electrospray ionization (ESI) mode. Within 12 min, eight quaternary ammonium drugs, each at 1 ng/mL in horse urine, could be detected. The confirmation of these drugs in urine samples was achieved by capillary electrophoresis tandem mass spectrometry (CE-MS/MS). A direct comparison of this method was made with existing liquid chromatography/mass spectrometry (LC-MS) methods in the detection and confirmation of glycopyrrolate and ipratropium bromide in horse urine. While the two drugs could be detected within the same CE-MS run at 1 ng/mL in urine, they could only be detected in separate LC-MS runs at 5 ng/mL in urine. In addition, CE-MS consumed a much smaller volume of extract; the analyte peak widths, in some cases, were much narrower; and as the quaternary ammonium ions were well separated electrophoretically from the mainly neutral urine matrix, a much cleaner background in the CE-MS total ion trace was observed. PMID- 11504054 TI - Binding studies of porphyrins to human serum albumin using affinity capillary electrophoresis. AB - The present work demonstrates that affinity capillary electrophoresis (ACE) can be employed as a valuable and powerful tool for studying the interactions between porphyrins and proteins in biological and biomedical research, such as the development of porphyrins and related compounds as efficient and selective photosensitizers in the photodynamic therapy of cancers. Binding constants of human serum albumin (HSA) to four biological porphyrins (uroporphyrin I, heptacarboxylporphyrin, coproporphyrin I, protoporphyrin IX), which possess a wide range of hydrophobicity, were estimated by ACE. Based on 1:1 molecular association between these individual porphyrins and HSA, the change of the electrophoretic mobility of HSA as a function of porphyrin concentration in the run buffer was measured and the binding constants were calculated from the slope of the Scatchard plots. The binding constant values were found to be 8.80 +/- 0.51 x 10(4) M(-1), 2.39 +/- 0.16 x 10(5) M(-1), 1.61 +/- 0.11 x 10(6) M(-1), and 9.34 +/- 0.30 x 10(6) M(-1) for uroporphyrin I, heptacarboxylporphyrin, coproporphyrin I, and protoporphyrin IX, respectively, and most of these results are in good agreement with those reported in the literature using conventional methods for binding measurements. Additionally, experimental binding constant data obtained using ACE was found to exhibit very good correlation with theoretical hydrophobicity values calculated using the Rekker's hydrophobic fragmental constant method, thus further supporting the hypothesis that the hydrophobicity of the porphyrin side chains play an important role in governing the hydrophobic interaction of porphyrins with serum proteins such as HSA. PMID- 11504055 TI - Capillary electrophoresis of high-molecular chitosan: the natural carbohydrate biopolymer. AB - Due to its high resolving power and diverse application range, capillary electrophoresis (CE) has been successfully applied to the analysis of carbohydrates. In this paper, a method for the determination of high-molecular chitosan (Mr 200,000) using CE is presented. We studied the optimal condition of buffer pH and type, and column type for determination of chitosan. Optimal CE performance was found when employing 100 mM triethylamine (TEA)-phosphate buffer, pH 2.0 and untreated fused-silica capillary (50 microm x 27 cm) for the chitosan analysis. Under optimum conditions, excellent linear responses were obtained in the concentration range of 1.25-20 microM, with a linear correlation coefficient of 0.9983. The standard deviations of the migration time and peak area were found to be 2.5 and 6.4%, respectively. This method could be readily applied to chitosan determination in real biological samples and commercial products. PMID- 11504056 TI - Separation of stilbenes by capillary electrophoresis and high-performance liquid chromatography. AB - Stilbenes, fluorescence whitening agents (FWAs), are usually added to cleaning agents in household and in industry. Capillary electrophoresis (CE) was often applied to separate various compounds simultaneously for its multinomial advantages. In this paper, we established analytical methods of six diaminostilbenes with CE and ion-pair chromatography (IPC). The optimum mobile phase for IPC was 11.78 mM tetrabutylammonium hydrogen sulfate (TBA) aqueous and acetonitrile. An IPC method has been developed for simple and direct separation for diaminostilbenes, anionic substances, with TBA as ion-pair reagent. Satisfactory linear ranges (7.0 x 10(-3) approximately 3.0 x 10 microg/mL), correlation coefficients (0.9992-0.9999), and detection limits (6-13 ng/mL) were obtained. Separations were also performed by capillary zone electrophoresis (CZE) using a buffer consisting of Tris (pH 10.1), n-tetradecyltrimethylammonium bromide (TTAB) and acetonitrile. A linear range of 5.0 x 10(-1) - 4.0 x 10 microg/mL, correlation coefficients between 0.9975 and 0.9998, and detection limits between 337 and 446 ng/mL were obtained. In particular, the separation of a pair of similar compounds (mass difference of 2) was achieved by addition of TTAB. The optimum analytical methods of CE and high-performance liquid chromatography (HPLC) were applied to commercial household with direct analysis and standard addition. No significant bias were shown between them by t-test at 95% confidence level. PMID- 11504057 TI - Determination of sialic acids in the serum of cancer patients by capillary electrophoresis. AB - A novel method for the determination of N-acetylneuraminic acid (NANA) and N glycolylneuraminic acid (NGNA) was developed by using high-performance capillary electrophoresis (HPCE) with UV detection at 195 nm. NANA and NGNA were separated directly and analyzed without pre- or postcolumn derivation. The detection limit of NANA is 9.6 x 10(-6) mol L(-1) and for mass 3.879 x 10(-14) mol (39 fmol). This method was applied for the determination of NANA in 30 normal human and 72 cancer patients. The results demonstrated that NANA in the sera of cancer patients increased significantly as compared with the normal human (P < 0.001). The new method is simple and sensitive, and is suitable for basic research and clinical application to malignant tumors. PMID- 11504058 TI - Determination of aristolochic acids in medicinal plants (Chinese) prepared medicine using capillary zone electrophoresis. AB - Aristolochic acids (I and II) are commonly found in medicinal plants such as Radix aristolochiae and have been reported to cause acute hepatitis and end-stage renal failure. The aim of this work was to develop a method for the analysis of aristolochic acids in medicinal plant/Chinese prepared medicine (CPM) using (CZE). The buffer used was 30 mM sodium tetraborate at pH 9.5, detection was at 254 nm, applied voltage at 18 kV and the temperature was set at 25 degrees C. The effect of ionic strength, pH, and applied voltage on the separation was investigated. The precision values (relative standard deviation, RSD, %) for the relative migration time and peak area or peak height for aristolochic acids I and II were found to be less than 0.3% and between 2.6 to 4.0%, respectively. The limit of detection for aristolochic acids I and II was found to be 1.2 and 0.9 mg/L, respectively. The proposed method using pressurized liquid extraction (PLE) with CZE was used to determine the amount of aristolochic acids in medicinal plants or CPM samples with complex matrix and the results were compared with high performance liquid chromatography (HPLC). Method precision (RSD, n = 6) was found to be less than 4% when those from applied to medicinal plants and CPM samples. PMID- 11504059 TI - Application of capillary electrophoresis for organic acid analysis in herbal studies. AB - A capillary electrophoresis (CE) procedure has been developed for the separation of 25 inorganic and organic acid anions using a buffer system consisting of 15 mM tris(hydroxymethyl)aminomethane, 3 mM 1,2,4-benzenetricarboxylic acid, 1.5 mM tetraethylenepentaamine (TEPA) and 20% methanol with pH adjusted to 8.4. A good separation of organic acids extracted from a mixture of Chinese traditional medicine (TCM) containing three herbs, Flos chrysthemi, Spica prunellae, and Folium mori was obtained using the procedure developed with satisfactory working range (0.20-77 mg/g), low detection limit (90-190 microg/g), and good repeatability (relative standard deviation 4.47-6.99%, n = 4). A satisfactory extraction of organic acids was achieved within 20 min using 0.1 M NaOH. The addition of TEPA to provide a reduced electroosmotic flow (EOF) environment was shown to remove interfering organic compounds extracted from TCM. The applicability of using organic acids as markers for determining the mixing ratio of constituent herbs for a TCM mixture was investigated using a three-component mixture with a 1:1:1 mixing ratio. A satisfactory mixing ratio of 1.04:1.09:0.98 was obtained using the methodology developed based on organic acids as markers. The application of our method for determining more complicated TCM mixtures has been discussed. PMID- 11504060 TI - Determination of oxoanions in river water by capillary electrophoresis. AB - A new capillary electrophoresis (CE) procedure was developed for simultaneous determination of ten oxoanions (CrO4(2-), SeO4(2-), MoO4(2-), WO4(2-), VO4(3-), SeO3(2-), As04(3-), TeO3(2-), TeO4(2-), and AsO3(3-)) which were baseline separated from each other and from the interfering UV absorbing anions (NO3- and NO2-) commonly found in environmental water samples. The new background electrolyte system developed contained 5 mM potassium phosphate and 0.007 mM octadecyltrimethylammonium hydroxide, pH 11.2. The optimized working conditions were electrokinetic sampling at -5 kV for 10 s, running voltage at -15 kV with 5 microA current, and detection wavelength at 205 nm. No interference was observed for non-UV-absorbing anions and UV-absorbing anions up to 20 and 10 times higher concentrations respectively. The speed of analysis was fast, with a complete CE run within 6 min. Wide linear ranges (1-2,000 microg/L), good repeatability in migration time (relative standard deviation RSD 0.55-2.8%), satisfactory precision in peak area (RSD 3.8-5.6%) and peak height (RSD 3.9-5.3%) measurement, and detection limits (1-25 microg/L) sufficiently sensitive to detect oxoanions found in environmental water samples were obtained. The reliability of the CE procedure developed had been established by recovery test and parallel method determination using atomic absoprtion spectrophotometry for real river water sample. PMID- 11504061 TI - Determination of pesticides in drinking water by micellar electrokinetic capillary chromatography. AB - A new analytical procedure using a two-step sample preconcentration (solid-phase extraction (SPE) and field-amplified sample stacking) prior to separation by micellar electrokinetic capillary chromatography was developed for the determination of 14 pesticides such as aldicarb, carbofuran, isoproturon, chlorotoluron, metolachlor, mecoprop, dichlorprop, MCPA, 2,4-D, methoxychlor, TDE, DDT, dieldrin, and DDE in drinking water. Good recoveries of pesticides were obtained using SPE with sample pH adjusted to 2-3. Field-amplified sample stacking was found to give enrichment factors up to 30-fold preconcentration of various pesticides under reversed polarity at -2 kV for 50 s. The optimized background electrolyte (BGE) consisted of 50 mM sodium dodecyl sulfate (SDS), 10 mM borate buffer, 15 mM beta-cyclodextrin (beta-CD), and 22% acetonitrile at pH 9.6, running was under 25 kV and detection at 202 nm. Good linearity was obtained for all pesticides with detection limits down to 0.04-0.46 ng/mL and a working range of 0.1-40 ng/mL. The repeatabilities of migration time and peak area were satisfactory with relative standard deviations (RSDs) between 0.66 and 13.6% and 4.1 and 28%, respectively. All pesticides except dieldrin were found to be detected at concentrations at least tenfold lower than the World Health Organization (WHO) guideline values. The analytical procedure developed offers an economic method for fast screening of multiple pesticide residues in drinking water for health protection. It had been applied to determine carbofuran and MCPA in agricultural run-off water samples, giving satisfactory repeatabilities of 10 and 12%, respectively, with n=5 for the determination of pesticides in contaminated water samples. PMID- 11504062 TI - Determination of carbonyl compounds in air by electrochromatography. AB - A new analytical procedure based on electrochromatography was developed for the separation and quantitation of 14 aldehydes and ketones (formaldehyde, acetaldehyde, acetone, acrolein, propionaldehyde, butanone, crotonaldehyde, isobutyraldehyde/butyraldehyde, 2-pentenaldehyde, isovaleraldehyde, valeraldehyde, benzaldehyde and hexanaldehyde) in ambient air currently to be regulated by the Hong Kong Environmental Protection Department. A volatile mobile phase using ammonium acetate compatible with mass spectrometry detection was developed and optimized as follows. Methanol: acetonitrile: aqueous buffer (4 mM ammonium acetate) = 65:5:30% v/v. With electrokinetic injection at 5 kV for 2 s, aqueous buffer pH adjusted to 8, applied voltage controlled at 25 kV, and detection at 360 nm in a fused-silica column packed with 3 microm ODS, a satisfactory separation was obtained for the 14 carbonyl compounds investigated. The working ranges in acetonitrile solution were found to vary from 0.25 to 79 mg/L with a correlation coefficient greater than 0.99, detection limits from 0.10 to 0.63 mg/L, and precision (relative standard deviation, n = 3) from 2.3 to 9.2%. Under an air flow rate of 0.3 L/min for a sampling time of 1 h, the working ranges varied from 0.030 to 11,000 microg/m3 and detection limits from 0.011 to 0.084 microg/ m3. The method has been successfully applied to monitor three carbonyl compounds in four urban and rural sites in Hong Kong and gave hourly readings of three carbonyl compounds for all the sites investigated with a separation time less than 25 min. PMID- 11504063 TI - Separation of dsDNA in the presence of electroosmotic flow under discontinuous conditions. AB - Separations of phiX-174/HaeIII DNA restriction fragments have been performed in the presence of electroosmotic flow (EOF) using five different polymer solutions, including linear polyacrylamide (LPA), poly(ethylene oxide) (PEO), hydroxypropylcellulose (HPC), hydroxyethylcellulose (HEC), and agarose. During the separation, polymer solutions entered the capillary by EOF. When using LPA solutions, bulk EOF is small due to adsorption on the capillary wall. On the other hand, separation is faster and better for the large DNA fragments (> 872 base pairs, bp) using derivative celluloses and PEO solutions. Several approaches to optimum resolution and speed by controlling EOF and/or altering electrophoretic mobility of DNA have been developed, including (i) stepwise changes of ethidium bromide (0.5-5 microg/mL), (ii) voltage programming (125-375 V/cm), (iii) use of mixed polymer solutions, and (iv) use of high concentrations of Tris-borate (TB) buffers. The DNA fragments ranging from 434 to 653 bp that were not separated using 2% PEO (8,000,000) under isocratic conditions have been completely resolved by either stepwise changes of ethidium bromide or voltage programming. Compared to PEO solutions, mixed polymer solutions prepared from PEO and HEC provide higher resolving power. Using a capillary filled with 600 mM TB buffers, pH 10.0, high-speed (< 15 min) separation of DNA (pBR 322/HaeIII digest, pBR 328/ Bg/l digest and pBR 328/Hinfl digest) has been achieved in 1.5% PEO. PMID- 11504064 TI - Microchip isoelectric focusing using a miniature scanning detection system. AB - A miniature scanning fluorescent detector has been developed for plastic microchannel isoelectric focusing (mIEF) analysis. The detector, comprised of a lamp and photomultiplier tube (PMT) on a moving stage, measured the real-time distribution of fluorescently labeled peptides subjected to gel-free mIEF. During the run, the effective length of the 6-cm channel was scanned every 9 s. Analysis was completed within 5 min while still obtaining high resolution and sensitivity. In addition, the scanning detector was used to characterize peptide migration properties within the channel by providing simultaneous temporal and spatial measurements. PMID- 11504065 TI - High-throughput polymerase chain reaction analysis of clinical samples by capillary electrophoresis with UV detection. AB - Routine genetic analysis of large numbers of individuals by polymerase chain reaction (PCR) using capillary electrophoresis is often restricted by the low throughput of standard protocols and the tedious sample preparation process. Here, we demonstrate that capillary electrophoresis with UV detection can be used in PCR-based DNA analysis starting from clinical samples without purification or complicated sample manipulation. After PCR reaction using cheek cells, blood, or HIV-1 gag DNA, the reaction mixtures were injected into a capillary array either on-line or off-line by base stacking. The use of multiplexed absorption detection and the elimination of any purification steps both before and after PCR reaction can potentially provide significant benefits compared to current methods for DNA analysis with regard to time, cost, and labor. PMID- 11504066 TI - Spatially focused deposition of capillary electrophoresis effluent onto surface enhanced Raman-active substrates for off-column spectroscopy. AB - Surface-enhanced Raman scattering (SERS) is employed to obtain distinctive spectra of compounds that are efficiently separated by capillary electrophoresis (CE) and deposited onto planar SERS-active substrates. A simple method is described that explains how to prepare SERS-active substrates by depositing a silver-colloid solution onto frosted-glass microscope slides, using a high efficiency nebulizer. Scanning electron micrographs reveal a layered coating of fairly uniform-sized, 100-nm silver nanoparticles with interstitial spaces ranging from a few to tens of nanometers. The on-column separation is monitored by laser-induced fluorescence, while electrofilament depositing the CE effluent onto a moving SERS-substrate. Subsequently, the SERS spectra and off-column electropherograms are obtained with a simple confocal Raman spectrometer. The test compounds used to demonstrate this technique include compounds of biological significance: benzyloxyresorufin, riboflavin, and resorufin. CE and Raman conditions are evaluated to determine their affects on the SERS signals. An average off-column efficiency of 100,000 plates/m and a signal reproducibility of 11% relative standard deviation were achieved. Characteristic spectra with major Raman bands exhibiting signal-to-noise ratios of greater than 3 were obtained for a 3.2-nL injection of 10(-6) M (706 fg) resorufin. Forming a self-assembled monolayer (SAM) on the substrate increases the sensitivity of the SERS technique and decreases the on-substrate broadening. Calibration plots for both plain- and SAM-SERS substrates are demonstrated. PMID- 11504067 TI - Electrically driven microseparation methods for pesticides and metabolites: V. Micellar electrokinetic capillary chromatography of aniline pesticidic metabolites derivatized with fluorescein isothiocyanate and their detection in real water at low levels by laser-induced fluorescence. AB - Anilines are important pollutants occurring in the environment as industrial discharges as well as the transformation products (i.e., metabolites) of a wide variety of commonly used pesticides. In this report, we describe the precolumn derivatization of anilines with fluorescein isothiocyanate (FITC) and their subsequent separation and detection by capillary electrophoresis-laser induced fluorescence (CE-LIF) detection. The FITC-aniline derivatives were readily detected at the 10(-10) M level. This limit of detection (LOD) was achieved in the presence of glycosidic surfactants complexed with borate at alkaline pH yielding the so-called in situ charged micelles. The glycosidic surfactants evaluated were n-octyl- and n-nonylglucopyranoside. Furthermore, and under optimum conditions, the FITC precolumn derivatization of the anilines was performed in real water (e.g., tap and lake water) spiked with anilines at the LOD level. The water matrices showed marginal effects on the extent of derivatization at the LOD level, and the possible interferents in the water samples did not affect the FITC-solute signal due to the selectivity of the derivatization and detection schemes. Besides filtration from microparticles, the real water samples did not necessitate extensive sample cleanup prior to derivatization. PMID- 11504068 TI - Electrically driven microseparation methods for pesticides and metabolites: VI. Surfactant-mediated electrokinetic capillary chromatography of aniline pesticidic metabolites derivatized with 9-fluoroenylmethyl chloroformate and their detection by laser-induced fluorescence. AB - In this report, we describe a surfactant-mediated electrokinetic capillary chromatography (SM-EKC) system for the separation of 9-fluoroenylmethyl chloroformate (FMOC)-derivatized anilines by capillary electrophoresis (CE). The SM-EKC system consisted of dioctyl sulfosuccinate (DOSS)/acetonitrile mixtures and was suited for the CE separation of the relatively hydrophobic FMOC-aniline analytes and other neutral compounds, e.g. alkylphenyl ketones. While the organic modifier acetonitrile (ACN) allowed the solubilization of the hydrophobic solutes and maintained the DOSS surfactant in its monomeric form by inhibiting micellization, the DOSS surfactant associated with the FMOC anilines to a varying degree thus leading to their differential migration and separation. Under these conditions, the FMOC-anilines were readily detected at the 10(-6) M level by UV at 214 nm and at the 10(-8) M level by laser-induced fluorescence (LIF) using a solid-state UV laser operating at 266 nm line as the excitation wavelength. The FMOC precolumn derivatization was also readily performed in lake water spiked with anilines at near the limit of detection (LOD) level. The lake water matrix showed no significant effects on the extent of derivatization at the LOD level as well as on the detection of the analytes due to the selectivity of the FMOC derivatization. The derivatization and detection of spiked lake water necessitated only the removal of microparticles by microfiltration prior to derivatization and detection. PMID- 11504069 TI - Determination of phycobiliproteins by capillary electrophoresis with laser induced fluorescence detection. AB - Phycobiliproteins are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae. They are composed of a protein backbone to which linear tetrapyrrole chromophores are covalently bound. Furthermore, they are water soluble highly fluorescent, and relatively stable at room temperature and neutral pH. For this reason, capillary electrophoresis-laser induced fluorescence (CE LIF) seems the idea method for determination of these important proteins. The effects of buffer additives such as sodium dodecyl sulfate (SDS)and putrescine on the separation of the three major phycobiliprotein types, namely allophycocyanin, phycocyanin, and phycoerythrin, with excitation and emission maxima at 652/660, 615/647, and 565(494)/575 nm, respectively, are considered. Detection limits for these proteins by CE-LIF are some 60-500 times better than by absorbance detection. The development of a fast and sensitive CE-LIF assay such as this is of potential significance to our understand ing of chemical and biological oceanographic processes. PMID- 11504070 TI - Capillary electrophoretic analysis of mu- and m-calpain using fluorescently labeled casein substrates. AB - Calpains are unique calcium-dependent thiol proteases that have been proposed to participate in a number of physiological processes including signal transduction and protein turnover in skeletal muscle. Calpains exist in two major forms. Interestingly, the two forms of protease show no significant difference in their action on various substrates. The only demonstrable difference in their activity involves the concentration of calcium required for activation. Both mu- and m calpains typically achieve half maximal activation at 50 microM and 0.7 mM calcium, respectively. The focus of this study was to examine the action of both forms of calpain on casein substrates and assess whether any differences could be observed in the resulting peptide finger print using capillary electrophoresis. Purified mu- and m-calpain were incubated for various lengths of time with Oregon Green labeled alphas- and beta-casein. The reactions were stopped with sodium dodecyl sulfate (SDS) and products separated by capillary electrophoresis in micellar electrokinetic capillary chromatography (MEKC) mode using laser-induced fluorescence (LIF) detection. Comparison of the electropherograms showed no difference in the peptide profile for either enzyme. However, it was found that beta-casein was hydrolyzed more extensively than alphas-casein, by both enzymes. Capillary electrophoresis was found to be a very sensitive technique for detection of calpain activity. Using beta-casein as substrate, the CE approach was able to detect 2-3 ng of calpain activity. The results also suggest that capillary electrophoresis is a useful tool for proteolytic investigations of protein structure. PMID- 11504071 TI - Diagnosis of late-infantile neuronal ceroid lipofuscinosis: a new sensitive method to assay lysosomal pepstatin-insensitive proteinase activity in human and animal specimens by capillary electrophoresis. AB - Batten disease, or human late-infantile neuronal ceroid lipofuscinosis (LINCL) is a familiar progressive degenerative disease affecting children, caused by a deficiency of a lysosomal proteinase (tripeptidyl peptidase I, TPP-I) and characterized by the accumulation of autofluorescent storage bodies in the brain and other tissues of the body. Current methodology used to diagnose this disease needs to be improved in order to have less invasive techniques with higher resolution and shorter assay time. In this report, we discuss the potential merits of micellar electrokinetic chromatography as an excellent tool that requires minute samples but offers high resolution and a short running time for monitoring TPP-I activity in human and animal specimens. PMID- 11504072 TI - Analysis of glutathione by capillary electrophoresis based on sample stacking. AB - Glutathione is a small peptide, which participates in cellular oxidation reduction and detoxification. It is present in most biological tissues at different concentrations. The oxidized and reduced forms of the peptide were measured in erythrocytes and myocardial tissue by capillary electrophoresis based on stacking. After tissue homogenization or hemolysis of the red blood cells, the samples were deproteinized with acetonitrile and injected filling about 13% of the capillary volume. The electrophoresis was performed at 10 kV using a separation buffer of 250 mM borate, 50 mM Tris, pH 8.0. Sample stacking increased the sensitivity of detection by 10-20-fold. PMID- 11504073 TI - Dopaminergic neurotransmission in psychiatry: should selection of patients only be based on DSM or ICD criteria? PMID- 11504074 TI - A multicentre observational study of radionuclide therapy in patients with painful bone metastases of prostate cancer. AB - A multicentre observational study was conducted by the Italian Association of Nuclear Medicine between 1996 and 1998. Twenty-nine Nuclear Medicine Departments participated. The aims of the study were to systematically evaluate the efficacy, toxicity and repeatability of radionuclide therapy of painful bone metastases (RTBM) in a large number of patients and to assess its incidence in patients with prostate cancer. Out of 818 treatments performed with a single i.v. dose of 148 MBq of strontium-89 chloride or 1,295 MBq of rhenium-186 hydroxyethylidene diphosphonate (HEDP), 610 could be evaluated (527 with 89Sr and 83 with 186Re HEDP). Eighty-one patients received multiple (up to five) RTBM. The total number of retreatments was 100. Patients were followed up for a period of 3-24 months. Results, assessed according to pain relief and consumption of analgesic drugs, were expressed at four levels: 1, no response; 2, mild response; 3, good response; 4, excellent response. Responses were: level 1 in 19%, level 2 in 21.3%, level 3 in 33.3% and level 4 in 26.4% of cases. Retreatments showed significantly (P<0.01) worse responses (48% levels 3+4), in comparison to first RTBM. Duration of palliation was 5.0+/-3.5 months, and was longer in cases of excellent response, in first RTBM, in patients with limited metastases and when 89Sr was used. Better responses were found in cases of limited skeletal disease, under good clinical conditions, when life expectancy exceeded 3 months, and in radiologically osteoblastic or mixed bone lesions. The only statistically significant predictive factor was life expectancy (P<0.001). Flare phenomenon (14.1% of cases) did not correlate with the response. Haematological toxicity (mild to moderate in most cases) mainly affected platelets, and was observed in 25.5% of cases overall and in 38.9% of retreatments. RTBM did not seem to prolong life, though in some cases scintigraphic regression of bone metastases was observed. The two radiopharmaceuticals did not show any statistically significant differences in palliative efficacy and toxicity, either in first RTBM or in retreatments. PMID- 11504075 TI - Monitoring the chemotherapeutic response in primary lung cancer using 99mTc-MIBI SPET. AB - Prediction and evaluation of the response to chemotherapy (CTx) are important for the correct and cost-effective treatment of patients with primary lung cancer. Although fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) is accepted as the most useful and accurate nuclear medicine technique for this purpose, its expense and limited availability restrict its use. Compared with PET agents, technetium-99m methoxyisobutylisonitrile (MIBI), which is used in nuclear oncology, is cheaper and available in any nuclear medicine clinic. With this in mind, in this study we aimed to evaluate the role of 99mTc-MIBI in monitoring the chemotherapeutic response in primary lung cancer. Twenty patients with primary lung cancer underwent 99mTc-MIBI single-photon emission tomography (SPET) at 15 min (early) and 3-4 h (delayed) after injection of the tracer. All patients underwent 99mTc-MIBI SPET study twice: before and after the 3rd cycle of CTx. Patients were divided into two groups, responders [R(+), n=10] and nonresponders [R(-), n=10], according to the change in tumour size on CT scan taken 2 weeks after the last cycle of the CTx. From the SPET images early and delayed tumour/lung ratios (ER and DR) were obtained before and after CTx. In the R(+) group, ER and DR decreased significantly after CTx, from 3.28+/-1.55 to 1.78+/ 0.72 (P<0.04) and from 3.23+/-1.55 to 2.0+/-0.88 (P<0.05), respectively. However, in the R(-) group, while ER showed a slight and statistically insignificant increase after CTx (from 2.51+/-1.23 to 2.65+/-1.86), DR increased significantly, from 2.74+/-1.37 to 3.27+/-2.31 (P<0.03). The percentage decreases in ER and DR in the R(+) group after CTx was significantly higher than that in the R(-) group: 34.36%+/-26.7% vs -13.78%+/-27.58% (P<0.0002) and 29.45%+/-25.23% vs -18.58%+/ 20.51% (P<0.0005), respectively. Using a decrease of > or =10% as a threshold for monitoring the chemotherapeutic response, 99mTc-MIBI had a sensitivity of 90% and a specificity of 100%. We found a positive correlation in 14 patients between ER and DR and survival: r=0.6754 and P=0.008, and r=0.5755 and P=0.031, respectively. Our results suggest that 99mTc-MIBI might be used in routine practice to monitor the chemotherapeutic response in patients with primary lung cancer, especially when PET is not available. PMID- 11504076 TI - Targeted bone marrow irradiation in the conditioning of high-risk leukaemia prior to stem cell transplantation. AB - Disease recurrence following stem cell transplantation (SCT) remains a major problem. Despite the sensitivity of leukaemias to chemotherapy and irradiation, conventional conditioning before SCT is limited by significant organ toxicity. Targeted irradiation of bone marrow and spleen by radioimmunotherapy may provide considerable dose escalation, with limited toxicity to non-target organs. In this study, 27 patients with high-risk or relapsing leukaemia were treated with rhenium-188-labelled CD66a,b,c,e radioimmunoconjugates (188Re-mAb) specific for normal bone marrow in addition to conventional conditioning with high-dose chemotherapy and 12 Gy total body irradiation prior to SCT. A mean activity of 10.2+/-2.1 (range 6.9-15.8) GBq 188Re-mAb was administered intravenously. Acute side-effects were assessed according to the CTC classification and patient outcome was determined. Mean radiation doses (Gy; range in parentheses) to relevant organs and whole body were as follows: 13.1 (6.5-22) to bone marrow, 11.6 (1.7-31.1) to spleen, 5.0 (2.0-11.7) to liver, 7.0 (2.3-11.6) to kidneys, 0.7 (0.3-1.3) to lungs and 1.4 (0.8-2.1) to the whole body. Stem cells engrafted in all patients within 9-18 days post SCT. Acute organ toxicity of grade II or less was observed. During follow-up for 25.4+/-5.3 (range 18-34) months, 4/27 (15%) patients died from relapse, and 9/27 (33%) from transplantation-related complications. Fourteen patients (52%) are still alive and in ongoing complete clinical remission. Radioimmunotherapy with the bone marrow-seeking 188Re labelled CD66 mAb can double the dose to bone marrow and spleen without undue extramedullary acute organ toxicity, when given in addition to high-dose chemotherapy and 12 Gy TBI before allogeneic SCT. This intensified conditioning regimen may reduce the relapse rate of high-risk leukaemia. PMID- 11504077 TI - Evaluation of head and neck cancer with 18F-FDG PET: a comparison with conventional methods. AB - The aim of this study was to evaluate the usefulness of 18F-FDG PET in the diagnosis and staging of primary and recurrent malignant head and neck tumours in comparison with conventional imaging methods [including ultrasonography, radiography, computed tomography (CT) and magnetic resonance imaging (MRI)], physical examination, panendoscopy and biopsies in clinical routine. A total of 54 patients (13 female, 41 male, age 61.3+/-12 years) were investigated retrospectively. Three groups were formed. In group I, 18F-FDG PET was performed in 15 patients to detect unknown primary cancers. In group II, 24 studies were obtained for preoperative staging of proven head and neck cancer. In group III, 18F-FDG PET was used in 15 patients to monitor tumour recurrence after radiotherapy and/or chemotherapy. In all patients, imaging was obtained at 70 min after the intravenous administration of 180 MBq 18F-FDG. In 11 of the 15 patients in group I, the primary cancer could be found with 18F-FDG, yielding a detection rate of 73.3%. In 4 of the 15 patients, CT findings were also suggestive of the primary cancer but were nonetheless equivocal. In these patients, 18F-FDG showed increased 18F-FDG uptake by the primary tumour, which was confirmed by histology. One patient had recurrence of breast carcinoma that could not be detected with 18F-FDG PET, but was detected by CT. In three cases, the primary cancer could not be found with any imaging method. Among the 24 patients in group II investigated for staging purposes, 18F-FDG PET detected a total of 13 local and three distant lymph node metastases, whereas the conventional imaging methods detected only nine local and one distant lymph node metastases. The results of 18F-FDG PET led to an upstaging in 5/24 (20.8%) patients. The conventional imaging methods were false positive in 5/24 (20.8%). There was one false positive result using 18F-FDG PET. Among the 15 patients of group III with suspected recurrence after radiotherapy and/or chemotherapy, 18F-FDG was true positive in 7/15 (46.6%) and true negative in 4/15 (26.6%). The conventional imaging methods were true positive in 5/15 (33.3%) and true negative in 4/15 (26.6%). One false negative (6.6%) and three false positive findings (20%) on 18F-FDG PET were due to inflamed tissue. The conventional imaging methods were false positive in three (20%) and false negative in three cases (20%). It is concluded that in comparison to conventional diagnostic methods, 18F-FDG PET provides additional and clinically relevant information in the detection of primary and metastatic carcinomas as well as in the early detection of recurrent or persistent head and neck cancer after radiotherapy and/or chemotherapy. 18F-FDG PET should therefore be performed early in clinical routine, usually before CT or MRI. PMID- 11504078 TI - The efficacy of technetium-99m ciprofloxacin (Infecton) imaging in suspected orthopaedic infection: a comparison with sequential bone/gallium imaging. AB - Technetium-99m ciprofloxacin (Infecton) has recently become established as a new radiopharmaceutical for the imaging of infection. This study was performed to determine the value of Infecton imaging in demonstrating orthopaedic infection and to compare the results with bone/gallium imaging. Twenty-two patients (12 female, 10 male; mean age 51.7+/-16.8 years) with suspected orthopaedic infective conditions were included in the study. The patients underwent three scintigraphic studies in the following sequence: 740 MBq 99mTc-methylene diphosphonate (MDP) three-phase bone scintigraphy; at least 2 days later, 370 MBq Infecton scan at 1 4 h; and finally, 185 MBq gallium-67 scintigraphy. 67Ga imaging could not be performed on four patients. All images were blindly interpreted by two independent observers. The final diagnosis was made by consensus when the readings were different. Interpretation of the early and late Infecton images was made separately, with visual findings being classified according to a four-grade scale (0, +, ++, +++). Images graded 0 and +, and also those regions which showed a decrease in uptake grade on late images as compared with early images, were classified as negative for infection; grades ++ and +++ were classified as positive. Bone/gallium images were considered positive when the images were spatially incongruent or when gallium uptake was more intense than that of 99mTc MDP. The diagnosis was confirmed by intraoperative microbiological or histological findings, or by the presence of gross purulence. The sensitivity of Infecton imaging was found to be 85%, the specificity 92% and the accuracy 88%, as compared to figures of 78%, 100% and 90%, respectively, for bone/gallium imaging. Although the two modalities showed a similar clinical yield, the easy availability of Infecton and the short investigation time make Infecton imaging the better option for the detection of orthopaedic infection. PMID- 11504079 TI - The optimal reference population for cardiac normality in myocardial SPET in the detection of coronary artery stenoses: patients with normal coronary angiography or subjects with low likelihood of coronary artery disease? AB - Both subjects with a low likelihood for coronary artery disease (CAD) and patients with normal findings on coronary angiography have been used as reference populations in non-invasive stress testing, including myocardial perfusion scintigraphy. Both of these criteria of normality--low likelihood of CAD and normal coronary angiography--have been criticised, and consensus on this issue is lacking. The aim of this study was to compare two different reference populations by testing the performance of artificial neural networks designed to interpret myocardial scintigrams. The networks were trained on myocardial perfusion scintigrams from 87 patients with angiographically documented CAD and on studies from one of two different reference groups: 48 patients with no signs of CAD based on angiography or 128 healthy volunteers with a likelihood for CAD <5%. The performance of the two different networks was then tested using scintigrams from a separate test group of 68 patients. Coronary angiography was used as the gold standard in this group. The network trained on patients with no signs of CAD based on angiography showed an area under the receiver operating characteristic (ROC) curve of 93%. The ROC area for the network trained on healthy volunteers was 72%, and this difference was statistically significant (P=0.03). The results of this study using artificial neural networks suggest that normal angiography should be preferred as the reference standard in myocardial scintigraphy when a patient is examined for CAD prior to possible angiography. Whether the same is true for other indications, e.g. in prognostic evaluation, is unknown. PMID- 11504080 TI - Somatostatin receptor sst1-sst5 expression in normal and neoplastic human tissues using receptor autoradiography with subtype-selective ligands. AB - Somatostatin receptors are known to be expressed in a large number of human tumours and represent the basis for in vivo tumour targeting. Stable somatostatin derivatives such as octreotide or lanreotide are the most frequently used radiopharmaceuticals acting through specific binding to somatostatin receptors; however, they do not bind with high affinity to all five receptor subtypes. Whereas the mRNAs for most receptor subtypes have been detected in tumours, it is in most cases unclear which of the receptor subtype proteins are expressed. Since in vitro receptor binding methods are close correlates and predictors of in vivo peptide receptor targeting, we took advantage of the recently developed subtype selective analogues and evaluated approximately 200 tumours for their receptor subtype protein expression in specific binding assays using autoradiography with 125I-[Leu8, D-Trp22, Tyr25]-somatostatin-28 and displacement by subtype-selective analogues. The majority of the tested neuroblastomas, meningiomas, medulloblastomas, breast carcinomas, lymphomas, renal cell carcinomas, paragangliomas, small cell lung carcinomas and hepatocellular carcinomas predominantly expressed sst2. The prostate carcinomas and sarcomas preferentially expressed sstl, while a majority of inactive pituitary adenomas displayed sst3 and, to a lesser extent, sst2. Growth hormone-secreting pituitary adenomas preferentially expressed sst2 and sst5; gastroenteropancreatic tumours and phaeochromocytomas frequently displayed sst2 and/or sstl. Non-neoplastic human tissues such as vessels, nerve plexus, pancreatic islets, prostatic stroma, adrenal medulla, spleen and germinal centres of the lymphoid tissues preferentially expressed sst2. However, the human gastric mucosa predominantly expressed sst1 while colonic mucosa displayed sst2. Interestingly, a minority of tumours showed a strong 125I-[Leu8, D-Trp22, Tyr25]-somatostatin-28 binding, of which less than 50% could be displaced by the sum of the five subtype-selective analogues. This observation suggests the existence of an as yet unknown subtype in selected tumours. This study is the first report to analyse the somatostatin receptor subtype expression in tumours with binding methods. We conclude that sst2, with high affinity for current radiopharmaceuticals such as Octreoscan, is predominantly expressed in a majority of tumours. Fewer tumour types (sarcomas, prostate cancers, inactive pituitary adenomas) preferentially express another subtype. This information is of importance with regard to the clinical applications and development of somatostatin analogues with distinct receptor subtype selectivities. PMID- 11504081 TI - Biodistribution of [11C] methylaminoisobutyric acid, a tracer for PET studies on system A amino acid transport in vivo. AB - [N-methyl-11C]alpha-Methylaminoisobutyric acid (11C-MeAIB) is a potentially useful tracer for positron emission tomography (PET) studies on hormonally regulated system A amino acid transport. 11C-MeAIB is a metabolically stable amino acid analogue specific for system A amino acid transport. We evaluated the biodistribution of 11C-MeAIB in rats and humans to estimate the usefulness of the tracer for in vivo human PET studies, for example, on regulation of system A amino acid transport and on tumour imaging. Healthy Sprague-Dawley rats (n=14) were killed 5, 20, 40 or 60 min after the injection of 11C-MeAIB, and the tissue samples were weighed and counted for 11C radioactivity. Ten lymphoma patients with relatively limited tumour burden underwent whole-body (WB) PET imaging with 11C-MeAIB. In addition, three other patients had dynamic PET scanning of the head and neck area, and the tracer uptake was quantitated by calculating the kinetic influx constants (Ki values) for the tracer. In animal studies, the highest activity was detected in the kidney, pancreas, adrenal gland and intestines. In humans, the highest activity was found in the salivary glands, and after that in the kidney and pancreas, similar to the results in animal studies. Rapid uptake was also detected in the skeletal muscle. In the graphical analysis, linear plots were obtained, and the mean fractional tracer uptake values (Ki) of the parotid glands (n=3) and cervical muscles (n=3) were 0.039+/-0.008 min(-1) and 0.013+/ 0.006 min(-1), respectively. The Ki value of the tumour (n=1) was 0.064 min(-1). Higher uptake of 11C-MeAIB into the tumour tissue was encountered. These results encourage further 11C-MeAIB PET studies in humans on the physiology and pathology of system A amino acid transport and on tumour detection. PMID- 11504082 TI - 3-[123I]Iodo-alpha-methyl-L-tyrosine uptake in cerebral gliomas: relationship to histological grading and prognosis. AB - 3-[123I]Iodo-alpha-methyl-L-tyrosine (IMT) is employed clinically as a tracer of amino acid transport in brain tumours using single-photon emission tomography (SPET). This study investigates the role of IMT SPET in the non-invasive histological grading and prognostic evaluation of cerebral gliomas. The files of patients investigated by IMT SPET in our clinic between 1988 and 1996 were evaluated retrospectively. Complete follow-up was available for 58 patients with cerebral gliomas investigated by IMT SPET shortly after tumour diagnosis. Seventeen patients had low-grade gliomas (WHO grade II), 14 had anaplastic gliomas (WHO grade III) and 27 had glioblastomas (WHO grade IV). Thirty-six cases were primary tumours and 22 cases, recurrences. Maximal and mean tumour-to-brain (T/B) ratios of IMT uptake at the first IMT SPET investigation were related to histological grading and survival time. Patients with low-grade gliomas showed significantly longer survival than patients with high-grade (grade III or IV) tumours. Gliomas without contrast enhancement on computed tomography or magnetic resonance imaging scans were associated with longer patient survival than tumours with contrast enhancement. The T/B ratios of IMT SPET showed no differences in relation to histological grading [WHO grade II: 1.73+/-0.59; WHO grade III: 1.74+/-0.38; WHO grade IV: 1.59+/-0.35, (mean+/-SD, T/B ratios of mean tumour uptake)]. The median survival time of patients with a high T/B ratio on IMT SPET was not significantly different from that of patients with a low T/B ratio (T/B ratio <1.6, 14.8 months; T/B ratio > or =1.6, 13.0 months). Thus, no evidence could be found for a relationship between IMT uptake in cerebral gliomas and either histological grading or survival time. Nevertheless, IMT SPET constitutes a useful method for the detection of primary and recurrent gliomas, determination of tumour extent and individual follow-up. PMID- 11504083 TI - Non-invasive methods for absolute cerebral blood flow measurement using 99mTc ECD: a study in healthy volunteers. AB - Radionuclide angiography with technetium-99m ethyl cysteinate dimer (ECD) allows non-invasive estimation of absolute cerebral blood flow (CBF), either by graphical Patlak-Gjedde analysis (PGA) or by spectral analysis (SA). Other methods estimate CBF by means of single-point arterial or venous sampling. The aim of this study was to evaluate radionuclide scanning and single-point venous sampling as potential clinical non- to minimally invasive methods for CBF determination in a large set of carefully screened healthy volunteers over the adult age range. Eighty-three carefully screened healthy volunteers (20-81 years, 43 males, 40 females) underwent planar radionuclide angiography with 925 MBq 99mTc-ECD. After correction for camera dead-time loss, hemispheric CBF was calculated from brain perfusion indices (BPI): BPI(G) for PGA and BPI(S) for SA. Of the volunteers, 49 also underwent venous sampling 6 min post injection, from which the lipophilic octanol extraction fraction and hemispheric brain fractionation index (BFI) were determined. All datasets were correlated and evaluated as a function of age and gender. Intrasubject variability for the BPI measurements was assessed in 11 volunteers by repeat study within 2 weeks of the first acquisition. Graphical and spectral analysis BPIs were strongly correlated (R=0.846, P<0.00001). This correlation coefficient increased to R=0.903 for the 74 cases in which graphical analysis was not hampered by temporal tracer retention in cervicobrachial venous valves. The BFI was weakly correlated to both BPI indices (BPI(G): R=0.34, P=0.02; BPI(S): R=0.31, P=0.04). The right hemisphere showed significant asymmetry for BPI(S) (AI=2.7%+/-4.3%, P<0.001), in correspondence with previous 99mTc-ECD data. BPI(G), BPI(S) and BFI were all inversely related to age, with an increased gradient after the age of 55 years, while there was no significant gender difference. The ratio of BPI(G) to BIP(S), which is a measure of the cerebral extraction fraction for 99mTc-ECD, was not dependent on age. Intersubject variability was 15.5% for both radionuclide scanning-based methods and 18.2% for venous sampling, and in all cases was independent of age. A much lower intrasubject variability was observed for BPI(S) (7.2%) than for BPI(G) (12.6%). This study provides reference values for normal perfusion indices assessed by graphical and spectral analysis. The results also indicate that spectral analysis allows the most reproducible estimate of hemispheric perfusion by means of an operator-independent and objective approach. Whereas accurate calibration of normal BPI(S) values to hemispheric CBF with established methods needs to be performed, non-invasive calculation of regional absolute CBF using 99mTc-ECD is possible by application of a linearisation algorithm. PMID- 11504084 TI - 99mTc-ECD brain perfusion SPET: variability, asymmetry and effects of age and gender in healthy adults. AB - Reliable and high-resolution reference data for regional cerebral blood flow measured with single-photon emission tomography (SPET) are necessary for optimal clinical and research use. Therefore, a large dataset of normal technetium-99m labelled ethylene cysteine dimer (ECD) perfusion SPET in carefully screened healthy volunteers with an age range spanning six decades was created, with correction for non-uniform attenuation and scatter and based on an anatomically standardised analysis. Eighty-nine healthy volunteers, stratified for gender (46 females, 43 males; age 20-81 years), were included. Twelve volunteers underwent repeated 99mTc-ECD SPET after 2.5+/-2.3 weeks. An automated whole-brain volume of interest analysis with MANOVA as well as voxelwise analysis using SPM99 was conducted. Average intersubject variability was 4.8% while intrasubject reproducibility was 3.0%. An age-related decline in tracer uptake was found in the anterior cingulate gyrus, bilateral basal ganglia, left prefrontal, left lateral frontal and left superior temporal and insular cortex (all P=0.001-0.02). There was an overall increase in right/left asymmetry with age, which was most pronounced in the frontal and temporal neocortex. The most significant correlations between AI and age decade were found in the prefrontal (R=0.35, P=0.001) and superior temporal neocortex (R=0.43, P<0.001). Women had significantly higher uptake in the right parietal cortex (P<0.001), while men showed higher uptake in the cerebellum and the left anterior temporal and orbitofrontal cortex (all P<0.01). This normative dataset allows age- and gender specific patient and group assessment of 99mTc-ECD perfusion SPET under a wide variety of clinical circumstances in relation to normal variations and highlights the importance of both age- and gender-specific normal datasets for optimal analysis sensitivity. PMID- 11504085 TI - Dynamic measurement of the flow rate in cerebrospinal fluid shunts in hydrocephalic patients. AB - We compared clinical outcomes in hydrocephalic patients and observed variation in the rate of flow in ventriculoperitoneal shunts with changes in posture in 231 separate examinations of shunt flow in 148 patients. A small cadmium telluride detector was placed over the shunt reservoir, and clearance of radioisotope injected into the reservoir was recorded as a measure of flow. Flow rate tended to increase during head elevation. Four patterns of radioisotope clearance were seen: type I, no flow; type II, adequate flow with moderate opening pressure; type III, adequate flow with low opening pressure; and type IV, excessive flow. This categorisation reflected clinical shunt function. Our method effectively assesses flow rate with the patient in a variety of postures or during movement, yielding useful information for adjustment of shunt valve pressure. PMID- 11504086 TI - Regional alterations of myocardial norepinephrine transporter density in streptozotocin-induced diabetic rats: implications for heterogeneous cardiac accumulation of MIBG in diabetes. AB - Cardiac scintigraphic studies using iodine-123 labeled metaiodobenzylguanidine ([123I]MIBG) have previously demonstrated the heterogeneous myocardial accumulation of radioactivity in diabetes. In this study, we investigated the myocardial regional distribution of [125I]MIBG and the effects of regional myocardial blood flow, myocardial norepinephrine (NE) content, and norepinephrine transporter (NET) function on regional [125I]MIBG accumulation in streptozotocin induced diabetic (STZ-D) rats. Dual-isotope autoradiographic studies using [125I]MIBG and technetium-99m labeled hexakis (2-methoxy-2-isobutylisonitrile) (99mTc-MIBI), a tracer for the measurement of myocardial blood flow, were carried out to investigate the changes in regional myocardial blood flow in STZ-D rats. Uptake of [125I]MIBG was similar between the anterior wall and the inferior wall in control rats. On the other hand, in STZ-D rats, uptake of [125I]MIBG in the inferior wall was significantly less than that in the anterior wall. Uptake of 99mTc-MIBI was not significantly different between the anterior and inferior walls in control or STZ-D rats, indicating that myocardial blood flow did not change regionally in either control or STZ-D rats, and that the blood flow was not responsible for the heterogeneity of the distribution of [125I]MIBG in STZ-D rats. In STZ-D rats, cardiac NE concentrations determined using an HPLC electrochemical detection (ECD) system were significantly increased in both the anterior and the inferior wall, although there was no significant difference in NE concentration between the anterior and inferior walls in control or STZ-D rats. Furthermore, the density and affinity of NET were investigated by studying the binding of [3H]desipramine to cardiac membranes. The Bmax values of the NET in the anterior wall were not significantly different between control and STZ-D rats, but the Bmax value of the NET in the inferior wall was significantly lower in STZ-D rats than in controls. In conclusion, myocardial MIBG uptake was reduced in the inferior wall of STZ-D rats compared with control rats; this decrease was correlated with the decrease in NET density, but was not dependent on the regional myocardial blood flow and NE concentration. These results suggest that regional fluctuations in NET levels in the inferior wall contribute to heterogeneous MIBG accumulation in diabetes. PMID- 11504087 TI - Monte Carlo evaluation of object shape effects in iodine-131 SPET tumor activity quantification. AB - In our clinical iodine-131 single-photon emission tomography (SPET) quantification for radioimmunotherapy, calibration and partial volume correction are based on measurements with phantoms containing spheres to simulate patient tumors even though real tumors are frequently nonspherical. In this study, Monte Carlo simulation was used to evaluate how object shape influences "spill-out" and "spill-in", which are major sources of quantification error associated with the poor spatial resolution of 131I SPET. Objects that varied in shape (spheres, cylinders, and an irregular structure) but were identical in activity and volume were simulated. Iterative reconstruction employed both attenuation and triple energy-window scatter compensation. VOIs were defined in the reconstructed images both using physical boundaries and using expanded boundaries to allow for the limited resolution. When physical boundaries were used, both spill-out and spill in were more significant for nonspherical structures than for spherical structures. Over the range of object volumes (50-200 ml) and at all background levels, VOI counts in cylinders were lower than VOI counts in spheres. This underestimation increased with decrease in object size (for the cold background 18% at 200 ml and -39% at 50 ml). It also decreased with increase in background activity because spill-in partially compensated for spill-out. It was shown that with a VOI larger than physical size, the results are independent of object shape and size only in the case of cold background. Activity quantification was carried out using a procedure similar to that used in our clinic. Quantification of nonspherical objects was improved by simple sphere-based partial volume correction, but the error was still large in some cases (for example, -39% for a 50-ml cylinder in a cold background and -35% for a 200-ml irregular structure defined on the basis of a typical tumor outlined on an X-ray computed tomography scan of a patient with non-Hodgkin's lymphoma). Partial volume correction by patient-specific Monte Carlo simulation may provide better quantification accuracy. PMID- 11504088 TI - Comparison of 99mTc-sestamibi lung/heart ratio, transient ischaemic dilation and perfusion defect size for the identification of severe and extensive coronary artery disease. AB - The ability to identify patients with severe coronary artery disease (CAD) by analysis of perfusion defects is limited. The lung/heart ratio (LHR) and transient ischaemic dilatation (TID) have been used for this purpose in thallium 201 scintigraphy. The value of these parameters in technetium-99m sestamibi single-photon emission tomography (SPET) imaging is controversial. In this study, therefore, we determined TID and LHR in a single-day rest/stress 99mTc-sestamibi SPET perfusion protocol and compared these measurements with perfusion defect size (PDS) and angiographic severity of CAD. Severe CAD was defined as >75% left main coronary stenosis and/or >90% proximal left anterior descending artery stenosis and/or >90% proximal stenosis in the left circumflex and right coronary arteries. LHR was determined from a stress anterior planar image recorded < or =6 min after exercise. TID ratio was derived from automatically calculated left ventricular rest/stress volumes, and PDS was measured based on semi-automated computer software (CEqual). Diagnostic accuracy and predictive values were compared between 22 patients with severe and 98 patients without severe CAD. LHRs showed a higher sensitivity (73%) for the assessment of severe CAD as compared to PDS and TID ratio (41% and 23% respectively, P<0.01), whereas specificity was highest for TID ratio [95%, P<0.01 when compared to PDS (84%) and LHR (82%)]. It is concluded that increased LHR in 99mTc-sestamibi myocardial perfusion imaging seems to yield good diagnostic accuracy in the detection of patients with severe CAD and may be derived from a single-day rest/stress study. PMID- 11504089 TI - Effects of ageing on serotonin transporters in healthy females. AB - The effect of ageing on brain serotonin transporters was evaluated in 19 healthy female volunteers (age range 22-74 years) using single-photon emission tomography and [123I]nor-beta-CIT. The study subjects were scanned 0.3, 3, 6 and 23 h after injection of 185 MBq of [123I]nor-beta-CIT. The ratio of the distribution volume for tracer in the midbrain to that in the cerebellum minus 1 was used as an index for serotonin transporter binding. An age-related decline of 2% per decade (r= 0.47; P<0.05) was found in the midbrain. The decline in [123I]nor-beta-CIT binding in the serotonin transporter-rich area is much less than that in dopamine transporters in the striatum (6% per decade). PMID- 11504090 TI - Radiation exposure and radiation protection of the physician in iodine-131 Lipiodol therapy of liver tumours. AB - Intra-arterial iodine-131 labelled Lipiodol therapy for liver cancer has been investigated for safety and efficacy over a number of years, but data on radiation exposure of personnel have remained unavailable to date. The aim of this study was to assess the radiation exposure of the physician during intra arterial 131I-Lipiodol therapy for liver malignancies and to develop appropriate radiation protection measures and equipment. During 20 intra-arterial administrations of 131I-Lipiodol (1110-1924 MBq), radiation dose equivalents (RDE) to the whole body, fingers and eyes of the physician were determined for (a) conventional manual administration through a shielded syringe, (b) administration with an automatic injector and (c) administration with a lead container developed in-house. Administration by syringe resulted in a finger RDE of 19.5 mSv, an eye RDE of 130-140 microSv, and a whole-body RDE of 108-119 microSv. The injector reduced the finger RDE to 5 mSv. With both technique (a) and technique (b), contamination of angiography materials was observed. The container allowed safe transport and administration of the radiopharmaceutical from 4 m distance and reduced the finger RDE to <3 microSv and the eye RDE to <1 microSv during injection. During femoral artery compression, radiation exposure to the fingers reached 170 microSv, but the whole-body dose could be reduced from a mean RDE of 114 microSv to 14 microSv. No more contamination occurred. In conclusion, radiation exposure was high when 131I-Lipiodol was administered by syringe or injector, but was significantly reduced with the lead container. PMID- 11504091 TI - Effect of corrections for blood glucose and body size on [18F]FDG PET standardised uptake values in lung cancer. AB - Standardised uptake values (SUVs) are commonly used as a semi-quantitative index of 2-[18F]fluoro-2-deoxy-D-glucose (FDG) tracer uptake in positron emission tomography (PET). Studies have shown that SUVs may depend on body size and blood glucose concentration and corrections for these effects have been proposed in the literature. This retrospective study investigated the effect of the proposed corrections on SUVs from a group of 154 patients with lung cancer who had scans on a dedicated PET scanner. A total of 252 SUVs were requested as an aid to staging during consideration for surgical resection. SUVs were calculated normalised to body weight (SUVw), lean body mass (SUV(LBM)) and body surface area (SUV(BSA)). The following correlations were examined: SUV with height, weight and body surface area for the different body size normalisations; SUVw and SUVw x blood glucose (SUV(BG)) with blood glucose; SUVw with scan time post injection; and SUVw with apparent lesion diameter. Significant correlations were only observed between: SUV(LBM) and height (P=0.007); SUVw and scan time (P=0.007); SUVw and lesion diameter (P=0.0005); and SUV(BG) and blood glucose (P<0.00001). The correlation between SUV(LBM) and height suggests that lean body mass as a function of height alone should not be used to normalise SUVs; however, the lean body mass calculated from a height and weight nomogram did not show this effect. The strong correlation between SUV(BG) and blood glucose concentration suggests that for non-diabetic fasted patients, lung tumour SUVs should not be adjusted for blood glucose. PMID- 11504092 TI - 99mTc-(V)DMSA scintimammography in the assessment of breast lesions: comparative study with 99mTc-MIBI. AB - The purpose of this study was to evaluate and compare the diagnostic accuracy of pentavalent technetium-99m dimercaptosuccinic acid [99mTc-(V)DMSA] and 99mTc methoxyisobutylisonitrile (MIBI) in the detection of primary breast cancer and metastatic lymph node involvement, and in the clarification of cases with indeterminate mammograms. Forty-one women (mean age+/-SD 55+/-7 years) referred for a suspicious breast lesion on physical examination and/or an abnormal mammogram underwent MIBI and (V)DMSA scintimammography (SMM) at separate sessions (48-h interval). Lateral prone and anterior supine images were obtained at 10 and 60 min after administration of 740-925 MBq of each tracer, in the arm contralateral to the breast lesion. The ipsilateral axillary region was also included in the field of view. The results of SMM and mammography were compared with histological findings. Breast cancer was histologically confirmed in 26 patients (mean diameter+/-SD 2.87+/-1.5 cm). Benign lesions were found in 15 patients (mean diameter+/-SD 2.04+/-2.7 cm). Mammography was definitely positive in 23/26 patients with breast cancer and indeterminate in 3/26 (sensitivity 88.4%). In benign lesions, mammography was true negative in 5/15 cases and indeterminate in 10/15 (specificity 33.3%). Both MIBI and (V)DMSA SMM detected 23/26 breast cancers (sensitivity 88.4%) and were true negative in 14/15 (specificity 93.3%). T/B ratios for breast cancer in MIBI and (V)DMSA scans were similar, and significantly higher than for benign lesions. MIBI correctly diagnosed 12/13 and (V)DMSA 11/13 cases in which the findings of mammography were indeterminate. In addition, (V)DMSA detected seven of eight cases of in situ ductal carcinoma (DCIS) associated with infiltrating carcinomas, while MIBI detected only two of these eight cases. (V)DSMA was also diffusely concentrated in benign lesions complicated by epithelial hyperplasia. Metastatic lymph node involvement was successfully imaged in 15/19 patients with metastatic disease by both agents (sensitivity 78.9%), while true-negative scans were observed in 19/22 (specificity 86.3%) patients with benign or malignant tumours without lymph node metastases. Linear regression analysis revealed a high coefficient of correlation between the (V)DMSA and the MIBI T/B ratios (r=0.8 P<0.001). We conclude that both (V)DMSA and MIBI show an excellent ability to detect breast cancer and its lymph node metastases. (V)DMSA also has a tendency to be diffusely and more intensely localised than MIBI in pre-invasive lesions, such as DCIS or epitheliosis, which are at risk of developing into malignancies. (V)DMSA could therefore provide a useful tool in the diagnosis of such lesions and possibly modify a predefined surgical plan. Finally, we believe that both tracers could offer an alternative method for elucidating nondiagnostic mammograms. PMID- 11504093 TI - Recent progress in fluorine-18 labelled peptide radiopharmaceuticals. AB - The application of biologically active peptides labelled with positron-emitting nuclides has emerged as a useful and interesting field in nuclear medicine. Small synthetic receptor-binding peptides are currently the preferred agents over proteins and antibodies for diagnostic imaging of various tumours. Due to the smaller size of peptides, both higher target-to-background ratios and rapid blood clearance can often be achieved with radiolabelled peptides. Hence, short-lived positron emission tomography (PET) isotopes are potential candidates for labelling peptides. Among a number of positron-emitting nuclides, fluorine-18 appears to be the best candidate for labelling bioactive peptides by virtue of its favourable physical and nuclear characteristics. The major disadvantage of labelling peptides with 18F is the laborious and time-consuming preparation of the 18F labelling agents. In recent years, various techniques have been developed which allow efficient labelling of peptides with 18F without affecting their receptor-binding properties. Moreover, the development of a variety of prosthetic groups has facilitated the efficient and site-specific labelling of peptides with 18F. The 18F-labelled peptides hold enormous clinical potential owing to their ability to quantitatively detect and characterise a wide variety of human diseases when using PET. Recently, a number of 18F-labelled bioactive peptides have shown great promise as diagnostic imaging agents. This review presents the recent developments in 18F-labelled biologically active peptides used in PET. PMID- 11504094 TI - Higher relative biological efficiency of alpha-particles: in vitro veritas, in vivo vanitas? PMID- 11504095 TI - Two years' experience using no-carrier-added meta-[123I]iodobenzylguanidine in clinical studies. PMID- 11504096 TI - Impact of patient positioning in scintimammography. PMID- 11504097 TI - Methodological considerations influence the clinical value of parathyroid localisation diagnostics. PMID- 11504098 TI - Ad astra per aspera: audacity and reinvention. PMID- 11504099 TI - Reinventing astronomy and our view of the Universe, again. PMID- 11504100 TI - Clinical manifestations and survival rates among patients with saccular intracranial aneurysms: population-based study in Olmsted County, Minnesota, 1965 to 1995. AB - OBJECTIVE: To report presenting clinical symptoms, previous medical history, and survival rates for people with saccular intracranial aneurysms (IAs), in a defined population. METHODS: The medical records of all residents of Olmsted County, Minnesota, with possible IAs were reviewed. Clinical manifestations at the time of diagnosis, previous medical history, demographic factors, and survival rates after diagnosis were determined. RESULTS: Of 270 people with IAs detected between 1965 and 1995, 188 exhibited symptoms at the time of diagnosis, including 74% of women and 63% of men (P = 0.054). Intracranial hemorrhage (ICH) was the most common presenting symptom (60% of all patients and 86% of patients who exhibited symptoms), followed by cranial nerve palsy, transient ischemic attacks, and seizures. Survival rates after detection (with the exclusion of cases that were first detected during autopsies) were dependent on the occurrence of ICH; 23% of patients who presented with ICH died by 1 day after diagnosis, compared with 5% of those who did not exhibit symptoms or exhibited symptoms but without ICH at presentation. At 5 years, 44.7% of patients with hemorrhage had died, compared with 29.4% of patients with symptoms other than hemorrhage. After the first 24 hours after detection, survival rates did not differ significantly for those presenting with or without hemorrhage. Predictors of better survival rates also included lower age and later calendar year of presentation. CONCLUSION: This study provides the first data on aneurysm characteristics, clinical symptoms, and survival rates among people with IAs in a defined population. During the study period, most aneurysms were detected in the context of an aneurysm-related symptom (particularly among women), with a large proportion of patients presenting with ICH. After the acute phase of hemorrhage, long-term survival rates among people with IAs were similar for those presenting with or without ICH. PMID- 11504101 TI - Management of cysts arising after radiosurgery to treat intracranial arteriovenous malformations. AB - OBJECTIVE: The proper treatment for patients with cyst formation after arteriovenous malformation radiosurgery is unknown. METHODS: The treatment of six patients who developed cysts after arteriovenous malformation radiosurgery is described. Four patients had undergone gamma knife radiosurgery (two patients developed cysts after repeat procedures), and two patients had undergone linear accelerator-based radiosurgery. The median prescription isodose volume at the time of the first radiosurgical procedure was 13.2 cm3 (range, 8.0-28.7 cm3). RESULTS: The cysts were discovered a median of 48 months (range, 24-89 mo) after radiosurgery. Three patients were originally without symptoms, and observation with serial imaging was performed; two of those patients developed symptoms 13 and 40 months later, whereas one patient has remained without symptoms for 51 months. Initial treatments for patients with symptomatic cysts included cyst aspiration (n = 3) and placement of a cystoperitoneal shunt (CPS) (n = 2). The median cyst volume was 14 cm3 (range, 4-63 cm3). Cyst recurrence occurred within 2 months for patients who underwent aspiration alone, necessitating placement of a CPS. Shunt placement eliminated the cysts for four patients, at a median follow up time of 16 months (range, 9-27 mo). One patient's cyst persisted despite CPS placement, and cyst excision was performed. No morbidity occurred with any of the cyst treatments. CONCLUSION: Cyst formation after arteriovenous malformation radiosurgery may occur many years after the procedure. Although most symptomatic cysts can be effectively treated with CPSs, cyst excision may be necessary if the mass effect is not relieved with the less invasive approach. PMID- 11504102 TI - Neurosurgical procedures in Jehovah's Witnesses: an increased risk? AB - OBJECTIVE: Because of the growing numbers of members worldwide in the sect of Jehovah's Witnesses, the refusal of blood and blood products due to religious reasons is increasingly encountered in clinical practice. As an alternative to blood transfusion, Jehovah's Witnesses accept blood-free volume substitution, and they sometimes accept the intraoperative reinfusion of autologous blood via a so called cell saver. The aim of this study was to examine whether the refusal of blood transfusion affects the surgical indications for neurosurgery and whether morbidity and mortality rates are higher after neurosurgical interventions in Jehovah's Witnesses. METHODS: The pre-, intra-, and postoperative hemoglobin and hematocrit values as well as coagulation parameters of a group of Jehovah's Witnesses (n = 103) were compared with those of a valid control group. RESULTS: The total intraoperative blood loss during spinal and intracranial surgery in Jehovah's Witnesses was often less than in controls, which suggests a less traumatic surgical procedure. Hemodynamically relevant blood loss occurred in two spinal and four intracranial interventions. The patients were managed without receiving blood transfusions or blood products, although increased time in the intensive care unit and increased convalescence days were necessary. Mean surgical times were 17.5 minutes longer for spinal interventions and 36.7 minutes longer for intracranial interventions than for patients in the control group. This may be attributed to a more careful and thus slower surgical technique and to longer and more extensive hemostasis. The length of hospitalization was 15% longer for Jehovah's Witnesses than for controls. CONCLUSION: The morbidity and mortality rates for Jehovah's Witnesses undergoing neurosurgery were not higher than those of the control group. Thus, it can be concluded that Jehovah's Witnesses did not have a higher risk when microsurgical techniques and extensive anesthetic monitoring were applied during neurosurgery. Because the surgical success rate for Jehovah's Witnesses corresponded to that of the control group, the increase in costs because of longer treatment times is compensated in the long run by avoiding a lengthier illness, sometimes with more expensive conservative therapy. PMID- 11504103 TI - Early proactive management of vestibular schwannomas in neurofibromatosis type 2. AB - OBJECTIVE: The treatment of patients with neurofibromatosis Type 2 has always been challenging for neurosurgeons and neurotologists. Guidelines for appropriate management of this devastating disease are controversial. METHODS: A retrospective study of 28 patients with neurofibromatosis Type 2 who underwent 40 middle fossa craniotomies for excision of their acoustic tumors is reported. Eleven patients underwent bilateral procedures. The study focused on hearing preservation and facial nerve results for this group of patients. The 16 male patients and 12 female patients ranged in age (at the time of surgery) from 10 to 70 years, with a mean age of 22.6 years. The mean tumor size was 1.1 cm (range, 0.5-3.2 cm), and the majority of tumors were less than 1.5 cm. RESULTS: Measurable hearing was preserved in 28 ears (70%), with 42.5% being within 15 dB pure-tone average and 15% speech discrimination score of preoperative levels. In 55% of cases there was no change in the hearing class, as defined by the American Academy of Otolaryngology-Head and Neck Surgery. Of the 11 patients who underwent bilateral operations, 9 (82%) retained some hearing bilaterally. After 1-year follow-up periods (mean, 12.8 mo), 87.5% of patients exhibited normal facial nerve function (House-Brackmann Grade I). CONCLUSION: Early surgical intervention to treat acoustic tumors among patients with neurofibromatosis Type 2 is a feasible treatment strategy, with high rates of hearing and facial nerve function preservation. PMID- 11504104 TI - Adrenocorticotropic hormone-producing pituitary tumors: 12- to 22-year follow-up after treatment with stereotactic radiosurgery. AB - OBJECTIVE: To study retrospectively long-term outcomes of patients with adrenocorticotropic hormone-producing pituitary tumors that were treated with stereotactic Leksell gamma knife unit radiosurgery. METHODS: Eighty-nine patients aged 5 to 67 years were treated between 1976 and 1985. Eighteen patients aged 18 to 68 years (mean age, 41 yr) were followed in detail. Fifteen patients were women. None had previously received conventional radiotherapy, but pituitary microsurgery had been performed in two patients, and one patient had had an adrenalectomy. In the remaining 15 patients, radiosurgery was the primary therapy. RESULTS: Sixty-four patients had one stereotactic treatment, and 25 patients had two or more treatments. No complications were observed during treatment and the immediate follow-up period. At follow-up, 17 patients had died 1 to 20 years after the first treatment. No deaths were related to the treatment. In our 18 patients, the follow-up time after the first radiosurgical treatment was 12 to 22 years (mean follow-up period, 17 yr). Urinary cortisol levels gradually normalized in 83% of the patients. No recurrences were observed. Pituitary hormone insufficiencies developed in about two of every three patients and occurred even more than 10 years after treatment. Eight patients had transient hyperprolactinemia. The patients' vision and visual fields were unaffected, and none of them had signs of radiation-induced side effects such as brain tumors or brain necrosis. CONCLUSION: Stereotactic radiosurgery is a safe and effective method in the treatment of patients with adrenocorticotropic hormone-producing pituitary tumors, and the effect of treatment is long-lasting. Stereotactic radiosurgery is mainly a complement to microsurgery because of its gradually appearing effect and the occurrence of pituitary insufficiency. New pituitary deficiencies may be found more than 10 years after treatment. PMID- 11504105 TI - Electrical stimulation of the prelemniscal radiation in the treatment of Parkinson's disease: an old target revised with new techniques. AB - OBJECTIVE: In the treatment of tremor and rigidity in patients with Parkinson's disease (PD), the prelemniscal radiation (RAPRL), a subthalamic bundle of fibers, is an exquisite target that can be visualized easily on ventriculograms. We sought to evaluate the effect of electrical stimulation of the RAPRL on symptoms and signs of PD in a long-term trial and to determine the localization of the stimulated area by means of stereotactic magnetic resonance imaging studies. METHODS: Ten patients with PD predominantly on one side had tetrapolar electrodes stereotactically oriented through a frontal parasagittal approach to the RAPRL contralateral to the most prominent symptoms. Preoperative and postoperative evaluations at 3, 6, 9, and 12 months after surgery were performed using conventional PD scales and quantitative evaluations of tremor amplitude and reaction time. Stereotactic high-resolution magnetic resonance imaging studies with the electrodes in place were used for anatomic localization. RESULTS: In all patients, temporary suppression of tremor occurred when the electrodes reached the target. The most effective stimulation was obtained when the pair of contacts was placed in the RAPRL. Long-term stimulation at 130 Hz, 0.09 to 0.450 milliseconds, and 1.5 to 3.0 V produced significant improvement in tremor and rigidity and mild improvement in bradykinesia. CONCLUSION: The RAPRL is an effective target for the alleviation of tremor and rigidity in patients with PD by either lesioning or neuromodulation; however, neuromodulation has the advantage of not inducing an increase in bradykinesia. The stimulated area seems to be independent of the subthalamic nucleus. PMID- 11504106 TI - Nonaccidental pediatric head injury: diffusion-weighted imaging findings. AB - OBJECTIVE: Diffusion-weighted imaging (DWI) reveals nonhemorrhagic posttraumatic infarction hours to days before conventional computed tomographic scanning or magnetic resonance imaging (MRI). We evaluated the diagnostic utility of DWI in children with nonaccidental head trauma. METHODS: The medical records and imaging examinations obtained between January 1998 and May 2000 for all children less than 2 years of age with presumed or suspected nonaccidental head injury were reviewed retrospectively. Twenty children who had undergone DWI within 5 days of presentation were included in the study. Computed tomographic scans, conventional MRI sequences, and DWI combined with apparent diffusion coefficient (ADC) maps were evaluated. RESULTS: Eleven girls and nine boys (median age, 5 mo) were studied. Eighteen children had presumed nonaccidental head trauma, and two children had suspected nonaccidental head trauma. Of the 18 children with presumed nonaccidental trauma, 16 (89%) demonstrated abnormalities on DWI/ADC, as compared with neither of the two children with suspected nonaccidental trauma. In 13 (81%) of 16 positive cases, DWI revealed more extensive brain injury than was demonstrated on conventional MRI sequences or showed injuries not observed on conventional MRI. DWI combined with ADC maps allowed better delineation of the extent of white matter injury. DWI/ADC abnormalities in the nonaccidental head injured children were likely to involve posterior aspects of the cerebral hemispheres, with relative sparing of the frontal and temporal poles. Severity on DWI correlated significantly with poor outcome (P < 0.005). CONCLUSION: DWI has broad applications in the early detection of infarction in children with nonaccidental head injury and enhances the sensitivity of conventional MRI. In the patients in this study, early DWI provided an indicator of severity that was more complete than any other imaging modality. The use of DWI may help to identify children at high risk for poor outcome and to guide management decisions. PMID- 11504107 TI - Surgical management of isolated hemangioblastomas of the spinal cord. AB - OBJECTIVE: Intramedullary hemangioblastomas are rare tumors, accounting for just 3% of all intraspinal neoplasms. The purpose of this study is to define the occurrence of isolated intramedullary hemangioblastomas and to analyze the role of the radiological studies and surgery for these lesions. METHODS: The charts of 19 consecutive patients operated on for isolated spinal intramedullary hemangioblastoma were reviewed. Preoperatively, all patients underwent magnetic resonance imaging and nine underwent spinal angiography. For all patients, the surgical approach was via posterior laminectomy. RESULTS: Our study sample comprised 6 women and 13 men, with an average age of 31.5 years (range, 16-75 yr). The mean prodrome was 20.8 months. Pain was the most common complaint. In all cases, the neoplasms were associated with a syrinx or edema. Gross total resection was achieved in all patients. At last follow-up examination (mean, 50.1 mo), 13 patients (68%) had improved and 6 patients (32%) had stabilized as compared with their preoperative clinical status. CONCLUSION: Isolated intramedullary hemangioblastomas typically have an indolent clinical course. These tumors have characteristic imaging properties on magnetic resonance imaging and angiography. Surgical removal of these lesions results in excellent long-term functional outcome. PMID- 11504108 TI - Long-term outcome of 104 patients after lumbar sequestrectomy according to Williams. AB - OBJECTIVE: The authors report the late outcome of 104 consecutive patients after Williams' sequestrectomy for virgin lumbar disc herniation. METHODS: The clinical records and the mailed questionnaires of 38 women and 66 men operated consecutively between March 1991 and November 1993 were analyzed retrospectively. In these 104 patients, 105 Williams' sequestrectomies were performed. RESULTS: The mean age at operation was 50.5 years (range, 23.2-86.7 yr), and follow-up ranged from 4.1 to 6.9 years (mean, 5.3 yr). Success rates, including excellent, good, and fair results, were 92.5%, 94.7%, and 93.3% for lumbalgia, radicular pain, and neurological dysfunction, respectively. Only a few patients did not improve or presented with worsened symptoms. Three of four patients with cauda equina syndrome recovered immediately after the intervention. There were eight (7.7%) minor postoperative complications, which were treated conservatively. Three women and three men (5.8%) underwent a revision procedure for a recurrent lumbar disc herniation at the same level after 0.4 to 3.1 years (mean, 1.8 yr). Two (1.9%) of these patients underwent further operations because of reherniation, and they required internal instrumentation eventually because of failed back surgery syndrome. CONCLUSION: Williams' conservative approach with sequestrectomy alone is a safe operative modality. It should be used whenever possible. As demonstrated in our series with a long follow-up time, the results are as favorable as or better than results after standard microsurgical lumbar discectomy with curettement of the interspace. Whether the incidence of failed back surgery syndrome can be reduced by this approach remains to be proved. PMID- 11504109 TI - Nuances in acoustic neuroma surgery. AB - This article describes surgical methods developed during my resection of 600 acoustic neuromas, 599 of which were total removals. This series began in 1965, with the introduction of the surgical microscope and the development of microsurgical technique. In the 1960s, there was no established technology for the microsurgery of acoustic neuromas. Much of the work of the great early neurosurgeons--Cushing, Dandy, Olivecrona, and other remarkable pioneers--did not really apply. New approaches, solutions, and tools were introduced at that time, and there was a long, steep learning curve. I hope that some of the discussion herein makes learning easier for the new generation of neurosurgeons as they develop their own operative nuances. PMID- 11504110 TI - Three-dimensional computed tomographic cranial base measurements for improvement of surgical approaches to the petrous carotid artery and apex regions. AB - OBJECTIVE: The bony and vascular anatomic features in the region of the petrous apex can vary significantly. These variations affect the operative view obtained via extended subtemporal or anterior transpetrosal approaches to cranial base lesions for individual patients. The goal of this study was to evaluate three dimensional computed tomography as a means of obtaining detailed preoperative anatomic information regarding bony and vascular landmarks and spatial relationships in the region of the petrous carotid artery and petrous apex. METHODS: We radiographically studied 15 patients (30 sides), using 0.8- to 1-mm thick, reconstructed, computed tomographic images. Special attention was given to the course of the petrous carotid artery. RESULTS: The petrous carotid artery was located lateral to the trigeminal impression. The size of the petrous apex medial to the horizontal petrous carotid artery was observed to be variable. The width of bone from the trigeminal impression to the wall of the internal auditory canal averaged 9.6 mm (range, 5.2-16.1 mm). A variable amount of bone overlying the internal auditory canal (4.5 mm) was also present. Multiple other relationships among key landmarks were quantified. CONCLUSION: There is significant variability in the anatomic features of the petrous apex among patients. For each patient, detailed preoperative information regarding the amount of bone to be removed during a cranial base procedure can be obtained using three-dimensional computed tomography. This information may be critical for determination of the amount of extra exposure that can be achieved via an anterior petrosectomy for each patient. PMID- 11504111 TI - Endoscopic endonasal approaches to the cavernous sinus: surgical approaches. AB - OBJECTIVE: After completion of an earlier endoscopic transsphenoidal anatomic study, we studied various endoscopic transsphenoidal approaches using cadaveric specimens to develop endoscopic endonasal surgical approaches to the cavernous sinus. METHODS: Ten cavernous sinuses in five artery-injected adult cadaveric heads were studied with 0-, 30-, and 70-degree angled 4-mm rod-lens endoscopes. The extent of the surgical exposure, the skewed endoscopic anatomic view, and the maneuverability of surgical instruments through their relative operating spaces were studied after various endoscopic endonasal approaches via one nostril. RESULTS: The paraseptal approach was used between the nasal septum and the middle turbinate and provided exposure at the anteromedial portion of the cavernous sinus. The contralateral paraseptal approach rendered a slightly more medial view at the cavernous sinus than did the ipsilateral approach. This approach offered limited surgical access to the lateral vertical compartment. The middle turbinectomy approach allowed surgical access to the lateral wall of the cavernous sinus, except for the superior orbital fissure and the orbital apex. The middle meatal approach, which was made between the middle turbinate and the lateral nasal wall, revealed the entire lateral vertical compartment of the cavernous sinus, including the orbital apex and the superior orbital fissure. However, its lateral tangential surgical trajectory and the absence of dedicated surgical tools limited the surgeon's surgical maneuverability. A combination of the middle turbinectomy and middle meatal approaches increased the operating space. CONCLUSION: Various endoscopic endonasal surgical approaches to the cavernous sinus were studied using adult cadaveric head specimens. PMID- 11504112 TI - Deep brain stimulator electrodes used for lesioning: proof of principle. AB - OBJECTIVE: Patients with chronically implanted deep brain stimulator (DBS) electrodes can encounter complications requiring hardware removal. We assessed the safety and efficacy of using implanted DBS electrodes to create a therapeutic lesion before their removal. METHODS: Revision or removal of the DBS electrodes was required in two patients who had previously undergone DBS implantation. We conducted a series of in vitro experiments to confirm that the DBS electrodes could be used to generate radiofrequency lesions and to assess the relationship between radiofrequency parameters and lesion size. With this information, and with the approval of the hospital ethical review board, implanted electrodes were used to create incremental radiofrequency lesions in the thalamus in one patient and in the subthalamic nucleus in another. The procedures were performed under local anesthesia with contiguous contacts of the DBS lead connected to the active and reference sites of the RF generator to create a bipolar lesion. RESULTS: A 51 year-old man with essential tremor and a thalamic DBS required repeated battery changes secondary to tolerance and high voltage demands. Rather than replacing the battery, a radiofrequency thalamotomy was performed by using the existing left DBS electrode. At the 6-month follow-up examination, successful lesioning provided near complete tremor control. A second patient, a 50-year-old man with Parkinson's disease who had undergone bilateral subthalamic deep brain stimulation, developed skin erosion over the DBS hardware. A subthalamic nucleus lesion was made through the right DBS electrode. Lesion position and size were confirmed with magnetic resonance imaging. CONCLUSION: Lesions can be made through chronically implanted DBS electrodes in a safe, graded fashion and can produce therapeutic benefit. PMID- 11504113 TI - Cellular engineering: molecular repair of membranes to rescue cells of the damaged nervous system. AB - PURPOSE: The acute administration of hydrophilic polymers (polyethylene glycol) can immediately seal nerve membranes, preventing their continuing dissolution and secondary axotomy. Polymer application can even be used to reconnect, or fuse, the proximal and distal segments of severed axons in completely transected adult mammalian spinal cord. CONCEPT: The sealing or fusion of damaged nerve membranes leads to a very rapid (minutes or hours) recovery of excitability in severely damaged nerve fibers, observed as a rapid return of nerve impulse conduction in vitro, as well as an in vivo recovery of spinal cord conduction and behavioral loss in spinal cord-injured adult guinea pigs. RATIONALE: Surfactant application produces a rapid repair of membrane breaches through mechanisms of interaction between the polymers and the aqueous phase of damaged membranes, and their ability to insert into, or seal, the hydrophobic core of the axolemma exposed by mechanical damage. DISCUSSION: This new technology applied to severe neurotrauma offers a clinically safe and practical means to rescue significant populations of spinal cord nerve fibers within 8 hours after damage--preventing their continued dissolution and secondary axotomy by secondary injury mechanisms. Application of this novel technology to other injuries to the peripheral and central nervous system is discussed, as well as a general application to soft tissue trauma. PMID- 11504114 TI - Alpha(v)beta3 and alpha(v)beta5 integrin expression in glioma periphery. AB - OBJECTIVE: This study analyzed the expression of integrins alpha(v)beta3 and alpha(v)beta5 in glioma tissue and focused on the periphery of high-grade gliomas. METHODS: The analysis was performed with Western blot, immunohistochemistry, and immunofluorescence, by use of two monoclonal antibodies able to recognize the functional integrin heterodimer. The expression of integrin related ligands and growth factors also was studied. Sections from the tumor periphery were classified as either tumor periphery (light tumor infiltrate or scant visible cells) or peritumor (heavy tumor infiltration). RESULTS: Our data on glioma tissues demonstrated that both integrins were expressed in glioma cells and vasculature and their expression correlated with the histological grade. Alpha(v)beta3 expression was prominent in astrocytic tumors. Both integrins were markers of tumor vasculature, particularly of endothelial proliferation. A high grade glioma periphery demonstrated a prominent expression of integrin alpha(v)beta3. Cells demonstrating alpha(v)beta3 positivity were identified as tumor astrocytes and endothelial cells by double imaging. The same cells were surrounded by some alpha(v)beta3 ligands and co-localized fibroblast growth factor 2. Matrix metalloproteinase 2 also was found to be co-localized with alpha(v)beta3 in the same cells. Alpha(v)beta3 expression was more relevant in tumor astrocytes. Alpha(v)beta3 integrin and vascular endothelial growth factor expression increased from the periphery to the tumor center. CONCLUSION: Our data support the role of integrins alpha(v)beta3 and alpha(v)beta5 in glioma associated angiogenesis. In addition, they suggest a role for integrin alpha(v)beta3 in neoangiogenesis and cell migration in high-grade glioma periphery. PMID- 11504115 TI - History of the surgical treatment of ulnar nerve compression at the elbow. AB - OBJECTIVE: Surgery to achieve ulnar nerve decompression at the elbow has been performed for nearly 2 centuries. Several methods have been developed, some of which have been abandoned. Historical insight improves understanding of current techniques and provides the basis for the development of new methods. Which treatment method is best is the topic of ongoing debate. METHODS: The literature was reviewed using the MEDLINE database. Standard textbooks and retrieved articles were checked for missing references. For older articles, the bibliographies of books and theses were consulted. When I was unsuccessful in finding information in standard biographies of authors of milestone publications, I contacted the hospitals or institutions in which these individuals worked or are currently working. RESULTS: A systematic chronological overview of the surgical treatment of ulnar nerve compression at the elbow is presented, with special attention to people who described a treatment method for the first time. CONCLUSION: This article is the first in the literature to provide information about and photographs of nearly all of the people who were important in the development of the surgical treatment of compression of the ulnar nerve at the elbow. PMID- 11504116 TI - Middle meningeal origin of the ophthalmic artery. AB - OBJECTIVE: To examine the microsurgical anatomy and clinical significance of an anomalous origin of the ophthalmic artery from the middle meningeal artery. METHODS: In the course of an anatomic study of the cavernous sinus, an anomalous ophthalmic artery arising from the middle meningeal artery was found. To further define the anatomy of the region, five additional skulls, in which the arteries and veins were injected with colored latex, were dissected using 3x to 40x magnification. RESULTS: The anomalous ophthalmic artery arose from the frontal branch of the middle meningeal artery, passed through the superior orbital fissure, and supplied the entire contents of the orbit, as well as giving rise to the central retinal artery. This study provides the first display of this anomaly in an anatomic dissection. CONCLUSION: The ophthalmic artery may infrequently arise from the middle meningeal artery. This anomaly places the ophthalmic artery at risk during procedures in which the dura is elevated from the greater and lesser wings of the sphenoid or when the sphenoid ridge is removed and during embolization procedures involving the branches of the external carotid artery. PMID- 11504117 TI - Radiosurgery-induced microvascular alterations precede necrosis of the brain neuropil. AB - OBJECTIVE: Radiosurgery is used as a therapeutic modality for a wide range of cerebral disorders. It is important to understand the underlying causes of deleterious side effects that may accompany gamma-irradiation of brain tissue. In this study, structural alterations in rat cerebral vessels subjected to gamma knife irradiation in vivo were examined, for elucidation of their potential role in necrosis formation. METHODS: A maximal center dose of 75 Gy was delivered to the rat parietal cortex with a 4-mm collimator, and changes occurring before necrosis formation were assessed 3.5 months after irradiation. Transmission electron microscopy, using horseradish peroxidase as a tracer, and scanning electron microscopy with vascular casting were performed. RESULTS: The capillary network in the irradiated area exhibited thickening and vacuolation of the basement membrane. The capillary density in the irradiated area was lower and the average capillary diameter was larger, compared with the nonirradiated side. These results indicate that substantial changes in the neuropil do not occur 2 weeks before the time of definite necrosis formation, whereas changes in the basement membrane are prominent. CONCLUSION: The necrotic response to intermediate doses of focused-beam irradiation appears after a considerable latency period and then progresses rapidly. This contrasts with previously reported responses to fractionated whole-brain irradiation, in which damage occurs slowly and gradually. Alterations in the microvascular basement membrane precede overt cellular changes in neuronal and vascular cells and provide an early index of cerebrovascular dysfunction in regions destined to undergo necrosis. PMID- 11504118 TI - Absence of early proinflammatory cytokine expression in experimental intracerebral hemorrhage. AB - OBJECTIVE: We sought to analyze the regional concentrations of proinflammatory cytokines in the acute period of intracerebral hemorrhage (ICH) and to test the hypothesis that ICH is associated with the expression of proinflammatory cytokines in the acute period. Although the expression of cytokines and their role in neuronal injury and inflammation is well characterized in cerebral ischemia and head injury, no information exists regarding expression of cytokines in ICH. METHODS: We introduced ICH in eight anesthetized mongrel dogs by autologous blood injection (6 ml) under arterial pressure in the deep white matter adjacent to the left basal ganglia. Samples of arterial blood and cerebrospinal fluid were collected, and tissue extracts were prepared from different regions of the brain for immunoassay of tumor necrosis factor alpha, interleukin (IL)-1beta, and IL-6 concentrations in animals with and without ICH. RESULTS: The tumor necrosis factor a levels (+/- standard error) in the cerebrospinal fluid 1 hour after ICH did not differ significantly between the ICH group and the control group (7.1 +/- 1.3 pg/ml versus 10.8 +/- 2.3 pg/ml, P = 0.22). Levels in the perihematoma region in the ICH group (96.6 +/- 3.1 pg/ml) were not significantly different from those in the control group (93.4 +/- 6.7 pg/ml, P = 0.7). IL-6 levels (+/- standard error) in the perihematoma region in the ICH group (116.3 +/- 13.3 pg/ml) did not differ significantly from those in corresponding regions in the control group (122.3 +/- 12.8 pg/ml, P = 0.7). IL 1beta levels were below 5 pg/ml in serum, cerebrospinal fluid, and extracts of different brain regions. CONCLUSION: The early pathophysiology of ICH does not involve significant expression of tumor necrosis factor a either in the perihematoma region or other regions of the brain. The observation suggests that the pathophysiology of ICH in the acute period is different from both cerebral ischemia and traumatic brain injury. PMID- 11504119 TI - Continuous intravenous infusion of CGS 26303, an endothelin-converting enzyme inhibitor, prevents and reverses cerebral vasospasm after experimental subarachnoid hemorrhage. AB - OBJECTIVE: Endothelin-mediated vasoconstriction has been implicated in the pathophysiology of cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH). Endothelin-1, the most potent vasoconstrictor peptide of the endothelin family, is synthesized initially as a large prepropeptide that requires multiple steps of post-translational processing for activation. The final step of this processing involves the proteolytic cleavage of a relatively inactive precursor, big endothelin-1, by the metalloprotease endothelin-converting enzyme. Previous findings have demonstrated that intravenous bolus injections of an endothelin converting enzyme inhibitor (CGS 26303) administered twice daily can prevent and reverse arterial narrowing in a rabbit model of SAH. However, attenuation of vasospastic response was incomplete and required relatively high doses to be effective in reversing vasospasm. Therefore, the present study evaluated an alternative protocol for administration of CGS 26303 to optimize the antispastic influence of this compound. METHODS: Continuous intravenous infusion of CGS 26303 at doses of 2.4, 8.0, or 24.0 mg/kg/d was initiated either 1 hour (prevention paradigm) or 24 hours (reversal paradigm) after experimental SAH in New Zealand White rabbits. All animals were killed by perfusion-fixation 48 hours after SAH. Basilar arteries were then removed and sectioned, and their cross-sectional areas were measured by use of computer-assisted video microscopy. RESULTS: Continuous intravenous infusion of CGS 26303 attenuated SAH-induced cerebral vasospasm in a dose-dependent manner in both the prevention and the reversal groups. These effects achieved statistical significance at all doses as compared with the SAH only or SAH-plus-vehicle groups. Furthermore, the attenuation of vasospasm after continuous infusion of CGS 26303 was more efficacious than that obtained with bolus injections. CONCLUSION: These findings provide further support for the use of endothelin-converting enzyme inhibition as a therapeutic strategy for reduction of cerebral vasospasm, and they also support the effectiveness of this strategy even when initiated after arterial narrowing has been established. The findings also indicate that continuous intravenous infusion of CGS 26303 is a more effective approach for attenuation of vasospasm than bolus intravenous administration. PMID- 11504120 TI - Inhibitory effects of sodium butyrate on proliferation and invasiveness of human glioma cells. AB - OBJECTIVE: Sodium butyrate (SB), a differentiation-inducing agent, has been demonstrated to inhibit cellular proliferation in a number of human cell lines. Its precise mechanisms remain to be clarified, however. We investigated molecular mechanisms of SB-induced growth suppression as well as the effects of SB on the invasiveness of human glioma cells. METHODS: Human glioma U87MG and U251MG cells were treated with 1 or 2 mmol/L SB for 48 hours, and the inhibition of cell growth was assessed by spectrophotometric analysis. Cell cycle analysis was carried out by the 5-bromo-2'-deoxyuridine incorporation method, and expression of cell cycle-regulatory proteins was determined by immunoblotting. In addition, invasiveness was assessed using a Transwell chamber (Iwaki, Tokyo, Japan) with extracellular matrix substrate fibronectin or laminin (Iwaki). RESULTS: SB treatment resulted in significantly suppressed proliferation of both U87MG and U251MG cells in a dose-dependent manner. It inhibited the G1-S transition, which was associated with increased expression of p21 and cyclin D1 and reduced pRb phosphorylation. Treatment with antisense oligonucleotide for Rb abrogated SB induced G1 arrest. p21 up-regulation was independent of the p53 status of the glioma cells. SB treatment also inhibited invasiveness on fibronectin and laminin. CONCLUSION: Our results indicate that SB may suppress the growth of human glioma cells through modulation of cell cycle progression and also may affect their invasiveness on extracellular matrix substrates, which suggests that SB may be a useful therapeutic agent in treating multiple aspects of malignant gliomas. PMID- 11504121 TI - 100 years of neurological surgery at the Mayo Clinic. AB - The practice of neurological surgery at the Mayo Clinic began as early efforts were undertaken by Drs. Charles Mayo and Emil Beckman, and surgical procedures were performed for trauma, infection, tumor, epilepsy, and hemorrhage. In 1919, the Section of Neurologic Surgery was established, with Alfred W. Adson as its first chair. Subsequently, Drs. Winchell McK. Craig, J. Grafton Love, Collin S. MacCarty, Ross H. Miller, and Thoralf M. Sundt, Jr., followed as eminent chairmen. Beginning with a modest number of cases per year, the neurosurgical service at the Mayo Clinic has grown to become one of the largest in North America. Under the current leadership of Dr. David G. Piepgras, approximately 3200 surgical procedures spanning the spectrum of subspecialties are performed each year by a staff of 10 neurosurgeons. This article traces neurosurgery at the Mayo Clinic, including several persons who contributed to its achievements over the past century. PMID- 11504122 TI - Metastatic hepatocellular carcinoma presenting as epidural hematoma: case report. AB - OBJECTIVE AND IMPORTANCE: A case of acutely symptomatic epidural hematoma caused by metastatic hepatocellular carcinoma (HCC) to the cranium is reported. This is a rare case of metastatic HCC without known primary presenting as an epidural hematoma. CLINICAL PRESENTATION: The patient presented with an acute onset of headache, aphasia, and right hemiparesis 2 weeks after he experienced minor trauma to the cranium. An emergency computed tomographic scan of the head revealed the presence of a left parietal epidural hematoma. INTERVENTION: An emergency evacuation of the epidural hematoma was performed, and metastatic HCC was diagnosed. CONCLUSION: The patient's neurological deficits were reversed with surgical intervention, and he is now undergoing palliative chemotherapy. This was the first clinical manifestation of HCC in this patient. This case reaffirms the neurosurgeon's role in the complex, multidisciplinary care of patients with craniospinal metastasis. PMID- 11504124 TI - Isolated, primary extranodal Hodgkin's disease of the spine: case report. AB - OBJECTIVE AND IMPORTANCE: To describe an unusual case of isolated, primary spinal Hodgkin's disease to draw attention to this disease as a possible diagnosis in patients with mixed inflammatory cell infiltrate lesions located in the spine. CLINICAL PRESENTATION: A 54-year-old African American woman presented with back pain and progressive lower-extremity weakness as a result of spinal cord compression from Hodgkin's disease of the thoracic vertebrae. INTERVENTION: The patient underwent decompressive surgery with stabilization, then was incorrectly treated with antituberculosis medication. When the disease progressed, later investigations revealed Hodgkin's disease, which responded to a course of radiation therapy. An extensive examination failed to detect lymphoma elsewhere. CONCLUSION: Isolated, primary extranodal Hodgkin's disease of the spine is extremely rare. Seven other cases were reported in 1954. Although the Hodgkin's disease in these cases may arise in the bone of the spine, the possibility of origin in the paraspinous soft tissues also cannot be excluded. PMID- 11504123 TI - Accumulation of intraventricular fat in an intracranial epidermoid tumor: case report. AB - OBJECTIVE AND IMPORTANCE: A fat component within the ventricles or subarachnoid space in fat-containing tumors such as an epidermoid or a dermoid has been observed in rare instances. However, there have been no reports regarding an increase in the size of such a fat component. We describe the case of an epidermoid tumor with intraventricular fat that showed an increase in size and amount. CLINICAL PRESENTATION: A 26-year-old woman was admitted with headache and diplopia. Computed tomography and magnetic resonance imaging of the brain revealed a fat-containing suprasellar tumor and widespread fat globules in adjacent sulci and cisterns and within the frontal horn of the lateral ventricle. INTERVENTION: The patient underwent a pterional craniotomy. Removal of the suprasellar tumor was nearly total. Histopathological examination revealed an epidermoid tumor. Sequential magnetic resonance imaging throughout the ensuing 65 month period revealed no evidence of tumor recurrence; however, the intraventricular fat remained and increased in size. The patient underwent surgery via the transcallosal approach at 69 months after the initial operation, and the presence of free-floating oily fat globules was confirmed. CONCLUSION: In the case of a fat-containing tumor with free fat in the cerebrospinal fluid spaces, careful serial examination is necessary, with particular attention to the possibility of changes in size. PMID- 11504125 TI - Conus perimedullary arteriovenous fistula with intracranial drainage: case report. AB - OBJECTIVE AND IMPORTANCE: Perimedullary arteriovenous fistulae (AVFs) do not commonly present with subarachnoid hemorrhage or intracranial venous drainage causing neurological symptoms. We present a case with both of these features. The patient was inadvertently treated for an unruptured intracranial aneurysm before his true problem was recognized. CLINICAL PRESENTATION: A 65-year-old man presented with sudden-onset lower-extremity weakness, diplopia, nausea, and dysarthria on the day of admission. A lumbar puncture documented subarachnoid hemorrhage, and imaging studies revealed a left middle cerebral artery aneurysm. It was noted during surgery that this aneurysm was unruptured, and the patient did not exhibit improvement after surgery. INTERVENTION: Spinal angiography demonstrated a spinal perimedullary AVF feeding from the left T12 radicular artery; venous drainage extended rostrally into the posterior fossa venous system. The AVF was surgically occluded via a posterior laminectomy at the level of the AVF. After surgery, the patient's symptoms began to abate. CONCLUSION: Conus perimedullary AVFs can have venous drainage that extends as far as intracranial veins, which can lead to confusing clinical findings because the symptoms may suggest an intracranial process, although the lesion is in the spine. Surgeons must be aware of this confusing presentation. PMID- 11504126 TI - Effects of surgical revascularization on peripheral artery aneurysms in moyamoya disease: report of three cases. AB - OBJECTIVE AND IMPORTANCE: The beneficial effects of surgical revascularization on rebleeding in moyamoya disease remain unclear. This report is intended to clarify the effects of surgical revascularization on peripheral artery aneurysms, which represent one of the causes of intracranial bleeding in moyamoya disease. CLINICAL PRESENTATION: Findings for three female patients who experienced intracranial bleeding are presented. Cerebral angiography revealed that intracranial bleeding resulted from the rupture of peripheral artery aneurysms arising from dilated collateral vessels such as the lenticulostriate artery. INTERVENTION: The patients successfully underwent superficial temporal artery middle cerebral artery anastomosis combined with encephaloduromyoarteriosynangiosis. Angiography demonstrated obliteration of the peripheral artery aneurysms, together with the disappearance or decrease in caliber of the parent collateral arteries, after surgery. None of the patients experienced rebleeding during the follow-up period (up to 52 mo). CONCLUSION: The results strongly suggest that surgical revascularization potentially improves cerebral circulation and decreases hemodynamic stress on collateral vessels, obliterating peripheral artery aneurysms. PMID- 11504127 TI - New method for obliterative treatment of an anterior wall aneurysm in the internal carotid artery: encircling silicone sheet clip procedure--technical case report. AB - OBJECTIVE AND IMPORTANCE: Aneurysms on the anterior surface of the internal carotid artery (ICA) have been shown to be somewhat different from ordinary berry aneurysms because they are rather small, grow rapidly in a short time, and easily lead to rupture, especially during surgery. The most difficult problem is that this type of aneurysm cannot be eliminated easily by an ordinary clipping procedure without causing apparent arterial stenosis or occlusion. CLINICAL PRESENTATION: A 52-year-old man experienced a subarachnoid hemorrhage because of a ruptured aneurysm located on the anterior surface of the ICA. The tiny aneurysmal body, which was covered with a layer of brain tissue, was successfully exposed. The ICA seemed to be atherosclerotic, and the aneurysmal portion was solitary and had a reddish color. TECHNIQUE: A large silicone sheet encircling clip (Vascwrap; Mizuho Ikakogyo Co., Ltd., Tokyo, Japan) was selected for this patient. The proximal margin of the silicone sheet was incised with a V-shaped cut, and the middle part of the sheet, which covered the diagonal part of the ICA, was trimmed to make it shorter. The blade of the fenestrated clip was applied to obliterate the aneurysm and was attached to the normal arterial wall together with this modified Vascwrap sheet to create a small space between the normal arterial wall and the surrounding Vascwrap sheet. Then tiny pieces of Teflon fiber (E.I. duPont de Nemours and Co., Wilmington, DE) was inserted from both margins, and the whole Vascwrap sheet was sealed with fibrin glue to ensure good adhesion. CONCLUSION: This method seemed adequate in treating this difficult aneurysm without causing postoperative regrowth or occlusion of the patient's ICA. PMID- 11504129 TI - Comparison of anatomic and neurophysiological methods for subthalamic nucleus targeting. PMID- 11504128 TI - Easy sellar reconstruction in endoscopic endonasal transsphenoidal surgery with polyester-silicone dural substitute and fibrin glue: technical note. AB - OBJECTIVE: To describe a simple method of sellar reconstruction after endoscopic endonasal transsphenoidal surgery that will allow rapid watertight closure of the sellar floor. METHODS: A bent sheet of a polyester-silicone dural substitute, fashioned for this purpose with scissors, is introduced into the sella after removal of the lesion. Because of the consistency of the sheet, it opens spontaneously and becomes stuck. Autologous fat tissue or gelatin foam is positioned thereafter, followed by another layer of the dural substitute; a film of fibrin glue completes the sealing. RESULTS: Fifteen patients underwent this method and no postoperative cerebrospinal leak or other complication was experienced. CONCLUSION: This easy method of sellar reconstruction represents an effective and fast possibility to perform the final step of the endoscopic transsphenoidal procedure, which otherwise may cause maneuverability problems in the limited space of one nostril. PMID- 11504130 TI - Preconquest Peruvian neurosurgeons: a study of Inca and pre-Columbian trephination and the art of medicine in ancient Peru. PMID- 11504131 TI - Near-real-time guidance using intraoperative magnetic resonance imaging for radical evacuation of hypertensive hematomas in the basal ganglia. PMID- 11504132 TI - Head protection in England before the First World War. PMID- 11504133 TI - Is there a benefit of preoperative meningioma embolization? PMID- 11504134 TI - Convulsive seizures following subdural application of fibrin sealant containing tranexamic acid in a rat model. PMID- 11504135 TI - Surgical experience with frontolateral keyhole craniotomy through a superciliary skin incision. PMID- 11504136 TI - Intraspinal clear cell meningioma: diagnosis and management--report of two cases. PMID- 11504137 TI - Prodromal signs and clinical factors influencing outcome in patients with intraventricular rupture of purulent brain abscess. PMID- 11504138 TI - Transorbital keyhole approach to anterior communicating artery aneurysms. PMID- 11504139 TI - Notice on use of AlloDerm. PMID- 11504140 TI - Dual diagnosis: HIV and mental illness, a population-based study. AB - This is a cross-sectional, population-based (n = 378,710) study using hospital discharge abstract data to determine the relative risk associated with having a dual diagnosis of mental illness and HIV/AIDS. The analysis addresses issues of gender, race, and age, as well as types of mental illness. Persons with a mental illness are 1.44 times more likely to have HIV/AIDS. Women are at increased risk of being dually diagnosed. There are no risk differences by race. Those with a specific diagnosis of substance abuse or a depressive disorder are more likely to have a diagnosis of HIV/AIDS. PMID- 11504141 TI - Co-occurring psychosocial distress and substance abuse in community clients: initial validity and reliability of self-report measures. AB - The current study examines the self-reports of 227 community support clients using a paper and pencil questionnaire that included the South Shore Problem Inventory-revised (a brief multidimensional psychosocial distress scale), a one item index of self-rated substance abuse (SRSA), a quantity-frequency index for alcohol consumption (QFI), and a one-item index measuring the frequency of marijuana use. Results support the factor structure and internal consistency of the SSPI-r, and show significant correlations among the substance use indices. Implications for including brief mental health and substance abuse measures are discussed within the context of routine assessment and practice evaluation. PMID- 11504142 TI - Community factors in the development of resiliency: considerations and future directions. AB - Researchers have questioned why some children and adolescents are more resilient than others in the face of adversity and have identified several protective factors. The present paper focuses on one of these variables, namely, support from caring adults in the community. We present a brief review of this component of the resiliency literature along with a discussion of some of the issues and challenges raised by the findings. It is suggested that the evidence is substantial enough and the possible rewards associated with exploiting these findings considerable enough to warrant mounting wide-scale community-based efforts to assist vulnerable youth. PMID- 11504143 TI - Physician unwitting participation in abuse and neglect of persons with developmental disabilities. AB - The vast majority of birth parents and foster parents for children with special needs do the best they can in challenging circumstances. Many describe the frustrating circumstance of knowing more than their physicians about their child's unusual medical conditions. In general, physicians are well-advised to listen carefully to the reports of observant, caring parents and foster parents, and to give these observations at least as much weight as their own. However, in the work of our traveling clinic we have observed a number of situations of abuse and neglect which were perpetrated by parents or foster parents, and where the actual acts of abuse and/or neglect were wholly or in part endorsed, in writing, by the treating physician. PMID- 11504144 TI - Adopting innovations--lessons learned from a peer-based hospital diversion program. AB - Moves to bridge the gap between research and practice have heightened interest in how service innovations are adopted. This paper reports on a peer-based hospital diversion program that provided short-term respite care, clinical monitoring, connection or re-connection to other mental health services, and peer support. The program was successful in providing services to the target population and was viewed as highly desirable by service recipients and clinical agencies. However, full adoption of this innovation was not realized and it closed barely a year after opening. Lessons learned from both the "life" and "death" of this program are offered. PMID- 11504145 TI - The role of the psychiatrist: job satisfaction of medical directors and staff psychiatrists. AB - In a previous survey of Columbia University Public Psychiatry Fellowship alumni, medical directors reported experiencing higher job satisfaction compared to staff psychiatrists. To further this inquiry, the authors conducted an expanded survey among the membership of the American Association of Community Psychiatrists (AACP). We mailed a questionnaire to all AACP members. Respondents categorized their positions as staff psychiatrist, program medical director or agency medical director, and rated their overall job satisfaction. The form also included a number of demographic and job characteristic items. Of 479 questionnaires mailed, a total of 286 individuals returned questionnaires (61%-12 forms were undeliverable). As in our previous survey, medical directors experience significantly higher job satisfaction compared to staff psychiatrists. Program and agency medical directors do not differ significantly. In addition, job satisfaction is strongly and negatively correlated with age for staff psychiatrists but not for medical directors. This survey strengthens the previously reported advantage medical directors have over staff psychiatrists regarding job satisfaction. The finding that job satisfaction decreases with increasing age of staff psychiatrists but not medical directors is particularly interesting, suggesting that staff psychiatrist positions may come to be regarded as "dead-end" over time. Psychiatrists are advised to seek promotions to program medical director positions early in their careers, since these positions are far more available, and provide equal job satisfaction, compared to agency medical director positions. PMID- 11504146 TI - Characterization and mechanisms of chromosomal alterations induced by benzene in mice and humans. AB - Elevated frequencies of chromosomal aberrations have been observed in the lymphocytes of benzene-exposed workers. Similar changes occurring in the bone marrow may play an important role in the development of leukemia. The objective of this research has been to characterize chromosomal alterations induced by benzene in mice and humans and to investigate the potential role of inhibition of topoisomerase II in the myelotoxic effects of benzene. The research is presented in three sections corresponding to the specific aims of the project: genotoxicity studies in the mouse, topoisomerase II studies, and initial studies using a new fluorescence in situ hybridization (FISH) approach to detect chromosome alterations in benzene-exposed workers. The results of the mouse experiments indicate that both chromosome breakage and aneuploidy are induced in the bone marrow of B6C3F1 mice following benzene administration. Chromosome breakage is the predominant effect, and this occurs primarily in the mouse euchromatin. Significant breakage within the mouse heterochromatin was also observed, as was aneuploidy. Breakage in the mouse bone marrow erythrocytes increased as a function of both dose and duration of benzene administration. The aneuploidy resulting from benzene exposure in mice was a relatively infrequent event, with increases of both chromosome loss and hyperdiploidy being observed. In the topoisomerase studies, benzene or its metabolites were shown to inhibit topoisomerase II enzyme activity in an isolated enzyme system, in a human bone marrow-derived leukemia cell line, and in vivo in the bone marrow of treated mice. The decreased activity was probably due to the rapid degradation of the topoisomerase II protein in the treated cells. In the human biomonitoring studies, the feasibility of using FISH with tandem DNA probes to detect chromosome alterations in interphase granulocytes and lymphocytes of benzene exposed workers was demonstrated. The results from the two worker studies were somewhat inconsistent, however. In the study of Estonian workers, characterized by lower exposures and a smaller sample size, the benzene-exposed workers exhibited elevated frequencies of breakage in the lq12 region as compared with those seen in controls. A suggestive trend toward increased hyperdiploidy was also seen, although the frequencies in the exposed workers were low and within the range of our laboratory's historical control frequencies. In the larger study of more highly exposed Chinese workers, no increase in breakage affecting the 1q12 region was seen among the exposed workers. A trend toward increased hyperdiploidy of chromosome 1 was seen in the exposed workers when the concentration of urinary benzene metabolites was used in conjunction with the frequency of hyperdiploidy observed in the lymphocytes of the individual workers. The results of these studies indicate that benzene exposure is characterized by chromosome breakage, primarily within the euchromatin, and modest increases in aneuploidy. These findings also provide the first direct evidence that benzene is capable of inhibiting the enzymatic activity of topoisomerase II in vivo, providing additional support for the hypothesis that inhibition of topoisomerase II contributes to benzene-induced toxicity and leukemogenesis. PMID- 11504147 TI - Biotransformation of MTBE, ETBE, and TAME after inhalation or ingestion in rats and humans. AB - The biotransformation of methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and tert-amyl methyl ether (TAME) was studied in humans and in rats after inhalation of 4 and 40 ppm of MTBE, ETBE, and TAME, respectively, for 4 hours, and the biotransformation of MTBE and TAME was studied after ingestion exposure in humans to 5 and 15 mg in water. tert-Butyl alcohol (TBA), a TBA conjugate, 2 methyl-1,2-propanediol, and 2-hydroxyisobutyrate were found to be metabolites of MTBE and ETBE. tert-Amyl alcohol (TAA), free and glucuronidated 2-methyl-2,3 butanediol (a glucuronide of TAA), 2-hydroxy-2-methyl butyrate, and 3-hydroxy-3 methyl butyrate were found to be metabolites of TAME. After inhalation, MTBE, ETBE, and TAME were rapidly taken up by both rats and humans; after termination of exposure, clearance from blood of the ethers by exhalation and biotransformation to urinary metabolites occurred with half-times of less than 7 hours in rats and humans. Biotransformation of MTBE and ETBE was similar in humans and rats after inhalation exposure. 2-Hydroxyisobutyrate was recovered as a major product in urine. All metabolites of MTBE and ETBE excreted with urine were eliminated with half-times of less than 20 hours. Biotransformation of TAME was qualitatively similar in rats and humans, but the metabolic pathways were different. In humans, 2-methyl-2,3-butanediol, 2-hydroxy-2-methyl butyrate, and 3 hydroxy-3methyl butyrate were recovered as major urinary products. In rats, however, 2-methyl-2,3-butanediol and its glucuronide were major TAME metabolites recovered in urine. After ingestion of MTBE and TAME, both compounds were rapidly absorbed from the gastrointestinal tract. Hepatic first-pass metabolism of these ethers was not observed, and a significant part of the administered dose was transferred into blood and cleared by exhalation. Metabolic pathways for MTBE and TAME and kinetics of excretion were identical after ingestion and inhalation exposures. Results of studies presented here suggest (1) that excretion of MTBE, ETBE, and TAME in rats and humans is rapid, (2) that biotransformation and excretion of MTBE and ETBE are identical in rats, and (3) that biotransformation and excretion of TAME is quantitatively different in rats and humans. PMID- 11504148 TI - Human cytochrome P450 isozymes in metabolism and health effects of gasoline ethers. AB - To reduce the production of carbon monoxide and other pollutants in motor vehicle exhaust, methyl tert-butyl ether (MTBE*), ethyl tert-butyl ether (ETBE), and tert amyl methyl ether (TAME) are added to gasoline as oxygenates for more complete combustion. Among them, MTBE is the most widely used. The possible adverse effect of MTBE in humans is a public concern, but the human enzymes responsible for metabolism of these gasoline ethers and the causes or factors for increased sensitivity to MTBE in certain individuals are totally unknown. This information is important to understanding the health effects of MTBE in humans and to assessing the human relevance of pharmacokinetics and toxicity data obtained from animals. In the present study, we demonstrated that human liver is active in metabolizing MTBE to tert-butyl alcohol (TBA), a major circulating metabolite and an exposure marker of MTBE. The activity is localized in the microsomal fraction but not in the cytosol. Formation of TBA in human liver microsomes is NADPH dependent and is significantly inhibited by carbon monoxide, which inhibits cytochrome P450 (CYP) enzymes. These results provide strong evidence that CYP enzymes play a critical role in the metabolism of MTBE in human livers. Human liver is also active in the oxidative metabolism of 2 other gasoline ethers, ETBE and TAME. We observed a large interindividual variation in metabolizing these gasoline ethers in 15 microsomal samples prepared from normal human livers. The activity level (pmol metabolite/min/mg) ranged from 204 to 2,890 for MTBE; 179 to 3,134 for ETBE; and 271 to 8,532 for TAME. The microsomal activities in metabolizing MTBE, ETBE, and TAME correlated highly with each other (r = 0.91 to 0.96), suggesting that these ethers are metabolized by the same enzyme(s). Correlation analysis of the ether-metabolizing activities with individual CYP enzyme activities in the human liver microsomes showed that the highest degree of correlation was with CYP isoform 2A6 (CYP2A6)+ (r = 0.94 for MTBE, 0.95 for ETBE, and 0.90 for TAME), which is constitutively expressed in human livers and known to be polymorphic. CYP2A6 displayed the highest turnover number in metabolizing gasoline ethers among a battery of human CYP enzymes expressed in human B lymphoblastoid cells. CYP2A6 coexpressed with human CYP reductase by a baculovirus expression system was also more active than CYP isoform 2E1 (CYP2E1) in the metabolism of MTBE, ETBE, and TAME. Kinetic studies on MTBE metabolism with human liver microsomes (n = 3) exhibited an apparent Michaelis constant (Km) of 28 to 89 microM and a maximum rate of metabolism (Vmax) of 215 to 783 pmol/min/mg. Metabolism of MTBE, ETBE, and TAME by human liver microsomes was inhibited by coumarin, a known substrate of human CYP2A6, in a concentration dependent manner. Monoclonal antibody against human CYP2A6 caused a significant inhibition (75% to 95%) of the metabolism of MTBE, ETBE, and TAME in human liver microsomes. Taken together, these results clearly indicate that, in human liver, CYP2A6 is a major enzyme responsible for metabolism of MTBE, ETBE, and TAME. Although CYP2E1 metabolizes diethyl ether and was previously suggested to be involved PMID- 11504150 TI - Clinician beliefs and practices related to increasing responsivity to the needs of Maori with alcohol and drug problems. AB - Culturally responsive treatments are often cited as essential for successfully addressing substance use-associated problems in indigenous and other ethnic groups. However, there has been little investigation of the support for this assertion among alcohol and drug-user treatment workers, or how it might translate into clinical practice. The current paper reports on the results of a survey of the New Zealand alcohol and drug-user treatment field, which canvassed these issues. Eighty-six percent of respondents advocated adjustment of clinical practice when working with Maori. Two key strategies were referral to specialist Maori groups or individuals and/or contacting/meeting with whanau (family). Comparisons were made between respondents who referred clients on and those who provided intervention themselves. Implications of results, limitations and future research are discussed. PMID- 11504149 TI - MTBE inhaled alone and in combination with gasoline vapor: uptake, distribution, metabolism, and excretion in rats. AB - The purpose of these studies was to extend previous evaluation of methyl tert butyl ether (MTBE)* tissue distribution, metabolism, and excretion in rats to include concentrations more relevant to human exposure (4 and 40 ppm) and to determine the effects of coinhalation of the volatile fraction of unleaded gasoline on the tissue distribution, metabolism, and excretion of MTBE. Groups of male F344 rats were exposed nose-only for 4 hours to 4, 40, or 400 ppm 14C-MTBE or to 20 or 200 ppm of the light fraction of unleaded gasoline (LFG) containing 4 or 40 ppm 14C-MTBE, respectively. To evaluate the effects of repeated inhalation of LFG on MTBE tissue distribution, metabolism, and excretion, rats were exposed for 4 hours on each of 7 consecutive days to 20 or 200 ppm LFG with MTBE (4 or 40 ppm) followed on the eighth day by a similar exposure to LFG containing 14C-MTBE. Subgroups of rats were evaluated for respiratory parameters, initial body burdens, rates and routes of excretion, and tissue distribution and elimination. The concentrations of MTBE and its chief metabolite, tert-butyl alcohol (TBA), were measured in blood and kidney immediately after exposure, and the major urinary metabolites-2-hydroxyisobutyric acid (IBA) and 2-methyl-1,2-propanediol (2MePD)-were measured in urine. Inhalation of MTBE alone or as a component of LFG had no concentration-dependent effect on respiratory minute volume. The initial body burdens of MTBE equivalents achieved after 4 hours of exposure to MTBE did not increase linearly with exposure concentration. MTBE equivalents rapidly distributed to all tissues examined, with the largest percentages distributed to liver. The observed initial body burden did not increase linearly between 4 and 400 ppm. At 400 ppm, elimination half-times of MTBE equivalents from liver increased and from lung, kidney, and testes decreased compared with the two smaller doses. Furthermore, at 400 ppm the elimination half-time for volatile organic compounds (VOCs) in breath was significantly shorter and the percentage of the initial body burden of MTBE equivalents eliminated as VOCs in breath increased significantly. These changes probably reflect a saturation of blood with MTBE at 400 ppm and strongly suggest that the uptake and fate of MTBE are notably different at exposure concentrations above and below 400 ppm. Single and repeated coexposure to 20 and 200 ppm LFG with MTBE had opposite effects on the total body burden of MTBE equivalents present at the end of exposures compared with those achieved after 4 and 40 ppm MTBE exposures: 20 ppm LFG increased and 200 ppm LFG significantly decreased the burdens of MTBE equivalents present. The effects of coexposure to LFG on blood levels of MTBE equivalents paralleled the effects on body burden. These differences in overall uptake of MTBE equivalents cannot be attributed to alterations of minute volume. The reason for the increase in overall uptake after 20-ppm LFG exposure is not clear. Decreased MTBE absorption (uptake) after single and repeated coexposure to 200 ppm LFG may be due to a decrease in solubility of MTBE in blood caused by inhalation of other hydrocarbons. Investigations on the blood/air partition coefficient of MTBE in the absence and presence of LFG would be needed to confirm this hypothesis. Single and repeated coexposure to either 20 or 200 ppm LFG significantly decreased the percentage of the initial body burden from MTBE equivalents in tissues, including liver, kidney, and testes, immediately and 72 hours after PMID- 11504151 TI - Whanaungatanga - a process in the treatment of Maori with alcohol- and drug-use related problems. AB - Maori, the indigenous people of New Zealand have advocated that culturally-based care requires goals and processes specifically defined within their ethnocultural context. Positive cultural identity and pride have been posited as integral to achieving wellbeing for Maori. A sample of Maori in treatment for alcohol- and drug-use associated problems was reported to believe that a sense of belonging to an iwi ("tribe") could contribute to the recovery process. Data relating to "tribal" affiliation, cultural self-identity and upbringing of that sample is presented. The key concepts of whanau ("family") and whanaungatanga ("relationships") in the treatment process for Maori are explored and the implications of their utility discussed. The findings highlight the complexity of providing treatment that is cognizant of ethnocultural factors. PMID- 11504152 TI - Use and misuse of benzodiazepines in Brazil: a review. AB - Benzodiazepines are among the most prescribed and consumed medication groups in the world. Although benzodiazepines are used in the treatment of several psychiatric and non-psychiatric disorders, and are generally safe and well tolerated, the potential for misuse and abuse is considerable. This makes the study and regulation of benzodiazepine prescription and consumption an item of concern in public health around the world. Most developed countries have consistent data of benzodiazepine sales and consumption; however, data from developing countries is scarce, making health policies on the use of benzodiazepines a much tougher issue in these countries. This article aims to review the epidemiology of benzodiazepine use in Brazil, as well as to analyze how legislation, physician misinformation and economic factors might contribute to making benzodiazepine abuse a problem in the country. PMID- 11504153 TI - An exploratory study of buprenorphine use in Bangladesh: a note. AB - A pilot study was carried out in Bangladesh during August and September, 1995, using a "snowball" technique with 30 male multiple drug users in order to investigate buprenorphine use, characteristics of the users, their reasons for its use and the drug's effects. PMID- 11504154 TI - Assessment of benzodiazepine dependence in alcohol and drug dependent outpatients: a research report. AB - In this study on 99 outpatients who were being treated for alcohol and/or drug dependence and also using benzodiazepines (BZDs), prevalence rates of DSM-III-R and ICD-10 substance dependence diagnoses were ascertained and scalability, reliability and validity of the scales of the Benzodiaepine Dependence Self Report Questionnaire (Bendep-SRQ) were assessed. The latter properties were investigated by Rasch analyses, discriminability coefficients, test-retest coefficients and factor analyses. BZD dependence was found to be a prevalent additional diagnosis. The psychometric findings appear to support the use of the Bendep-SRQ at outpatient addiction centers, which could contribute to a more differentiated treatment for poly-substance dependence. PMID- 11504155 TI - Drug-dependent women: demographic and clinical characteristics in a Brazilian sample. AB - This study compared baseline demographics, clinical characteristics and patterns of substance use of 99 substance misusing women seeking treatment in a Brazilian hospital who had been diagnosed with drug dependence (alcohol dependence, if present, was not the most important) against 162 women diagnosed with alcohol dependence. Most of the drug-dependent women in this study were dependent on cocaine (73.7%). It was found that drug-dependent women, at entry, were younger, with a higher educational level, single or lived alone, and had a job outside home more often than alcoholics; they also had less alcohol-use related problems in the family and more relatives with problems with other drugs. They sought treatment mainly by self-initiative and reported more past suicide attempts than their alcoholic counterparts. In addition, they began drug use at the same age but increased drug use, as well as seeking treatment, significantly earlier. Alcoholics had more psychiatric comorbidity. The findings point out heterogeneity among chemically-dependent Brazilian women. PMID- 11504156 TI - Adolescent marijuana use and adult occupational attainment: a longitudinal study from age 18 to 28. AB - This report uses data from a nationally representative longitudinal sample to examine the relationship of marijuana use by high school seniors to occupational attainment 10 years later. Analyses were conducted separately by gender, with and without controlling for other variables. Control variables, all measured when respondents were seniors, were: academic performance, educational aspirations, and occupational aspirations. Results indicate that the influence of marijuana use on occupational attainment is considerably different for males and females. PMID- 11504157 TI - New treasures from old? EPHESUS. Eplerenome Post-AHI Heart Failure Efficacy and Survival Study. AB - Impressive clinical benefits have been derived by inhibiting the renin angiotensin-aldosterone system with angiotensin-converting enzyme inhibitors. There is growing evidence that aldosterone plays a contributing role in the pathogenesis of heart failure beyond its sodium retention properties. EPHESUS, a trial of a novel aldosterone antagonist, eplerenone, in patients with myocardial infarction, left ventricular dysfunction and pulmonary congestion will determine whether this approach produces additive clinical benefits in modernly managed patients. PMID- 11504158 TI - The Brugada syndrome. PMID- 11504159 TI - Adeno-associated virus-mediated transfer of endothelial nitric oxide synthase gene inhibits protein synthesis of rat ventricular cardiomyocytes. AB - We investigated whether nitric oxide (NO) synthase gene transfer could attenuate growth of cultured cardiac myocytes. First, we investigated the effects of exogenous NO and cGMP analog on protein synthesis of cultured neonatal rat cardiac myocytes. The NO donor 3-morpholino-sydnonimine-hydrochloride (SIN-1) and 8-bromo-cGMP caused concentration-dependent decreases in phenylephrine-stimulated incorporation of 3H-leucine into cardiac myocytes. We then transferred endothelial constitutive NO synthase (ecNOS) gene into cultured neonatal rat cardiac myocytes using adeno-associated virus (AAV) vectors. ecNOS gene transfer into cardiac myocytes induced 140 kD ecNOS protein expression and significantly increased cGMP contents of myocytes compared with control cells. ecNOS gene transfer inhibited 3H-leucine incorporation into cardiac myocytes in response to phenylephrine, which was significantly recovered in the presence of the NOS inhibitor N(G)-monomethyl-L-arginine acetate. These results indicate that endogenously generated NO by ecNOS gene transfer using AAV vectors inhibits the alpha-adrenergic agonist-induced cardiac protein synthesis at least partially via cGMP production. PMID- 11504160 TI - Differential ability of human endothelial cells to internalize and express exogenous DNA. AB - Vascular endothelium gene expression regulates blood-vessel wall interactions, vascular permeability, smooth muscle cell growth and tone. The possibility to introduce exogenous DNA or RNA sequences in endothelial cells represents a novel therapeutic approach of vascular disease. The aim of the work was to investigate the ability of endothelial cells to internalize and express exogenous DNA sequences. Human umbilical vein endothelial cells (HUVEC) were transfected with either a 780 bp fluorescein-labeled DNA (FITC-DNA) or pEGFP-C1 plasmid encoding for a green fluorescent protein (GFP), using the cationic liposome DOTAP as transfection reagent. The transfected cell population was passed through a FACScan apparatus and percentage of fluorescent cells was determined using a FACScan analysis programme. The SW620 tumor-derived cell line was used as control. The percentage of FITC-DNA positive cells was 66.0% for HUVEC and 45.0% for SW620 cells. On the contrary, the percentage of GFP-positive cells was 13.8% and 43% for HUVEC and SW620, respectively. By increasing the amount of DNA as well as the protocol of administration the percentage of GFP-positive HUVEC was enhanced suggesting a rapid degradation of DNA in the HUVEC cytoplasm. PMID- 11504161 TI - Disopyramide and its metabolite enhance insulin release from clonal pancreatic beta-cells by blocking K(ATP) channels. AB - In an insulin-secreting pancreatic beta-cell line (MIN6), insulin release was caused by disopyramide, an antiarrhythmic drug with Na-channel blocking action, and its main metabolite mono-isopropyl disopyramide (MIP). Insulin secretion, measured as immunoreactive insulin (IRI), was accelerated to 265.7% of the control by disopyramide and to 184.4% by MIP, with half-effective concentrations (EC50) of 30.9 +/- 1.5 microM and 92.4 +/- 2.2 microM. We tested the possibility that these drugs induce insulin release by inhibiting ATP-sensitive K+ (K(ATP)) channels of MIN6 cells. In the cell-attached or ATP-free inside-out mode with patch membranes on MIN6 cells, K-selective channels were recorded with unitary conductance of 70.5 +/- 3.5 pS (150 mM external K+ ions at room temperature). The channels were concluded to be MIN6-K(ATP) channels because they were closed by extracellular high glucose (11.0 mM) or glibenclamide (200 nM) and were reversibly activated by diazoxide (50 microM). In the inside-out patch mode, they were inhibited by micromolar ATP. In both cell-attached and insideout mode, disopyramide and MIP inhibited single MIN6-K(ATP) channels. In the inside-out mode, they produced a dose-dependent inhibition of channel activity: the half blocking concentrations (IC50) were 4.8 +/- 0.2 microM for disopyramide and 40.4 +/- 3.1 microM for MIP. It was therefore concluded that both agents exert insulinotrphic effect through the inhibition of membrane K(ATP) channels in MIN6 cells. PMID- 11504162 TI - Effects of mibefradil, a T- and L-type calcium channel blocker, on cardiac remodeling in the UM-X7.1 cardiomyopathic hamster. AB - Abnormalities of T-type calcium channel function reported to occur in the transition phase to heart failure in the hamster cardiomyopathy may contribute to progression of the disease. We tested the hypothesis that chronic exposure to mibefradil, a selective T-type calcium channel blocker, improves the deleterious cardiac remodeling observed in this condition. In the present study, 40 normal (N) and 40 UM-X7.1 cardiomyopathic hamsters (CMH), aged 180 days, were treated daily by gavage for 21 days with mibefradil (30 mg/Kg). Eight to 10 animals from each group were sacrificed at the end of the treatment period while the remainder were followed for an additional 30 days without treatment (washout period). Hearts were harvested, fixed with 10%-buffered paraformaldehyde and then cut in half down the middle. Several slices were dehydrated, embedded in paraffin and stained with Masson Trichrome. Wall thickness and dilatation index of the left ventricle were estimated by planimetry. Myocardial capillary density was also computed. The greater heart weight/body weight ratio seen in untreated CMH (7.7 +/- 0.4 vs 5.5 +/- 0.2 in N, p < 0.05) was improved with mibefradil. The dilatation index averaged 0.504 +/- 0.04 in N was increased in untreated CMH (0.566 +/- 0.03) and ameliorated in mibefradil-treated CMH. The 1-month washout period led to further deterioration of the dilatation index in untreated and mibefradil-treated CMH. Capillary density averaged 10,000 +/- 781 per mm2 in hearts from untreated N hamsters and 8,830 +/- 795 per mm2 in untreated CMH (p = NS). Chronic exposure to mibefradil resulted in a significant reduction of capillary density in both N and CMH hearts. Following the 1-month washout period, the change in myocardial capillary density associated with mibefradil was no longer detectable. In conclusion, chronic exposure to mibefradil, a T- and L-type calcium channel blocker, exerts opposite effects during the transition phase to heart failure in CMH, improving the deleterious left ventricular remodeling in UM X7.1 hamsters and reducting myocardial capillary density independently of the disease process. PMID- 11504163 TI - Late coronary artery reperfusion has additive beneficial effects on infarct expansion when combined with early angiotensin converting enzyme inhibitor therapy post myocardial infarction. AB - BACKGROUND: Individually, late coronary artery reperfusion and early angiotensin converting enzyme (ACE) inhibitor therapy prevent infarct expansion post myocardial infarction (MI). OBJECTIVES: To examine the effect of late reperfusion on infarct expansion when added to early ACE inhibitor therapy post MI. METHODS: Rats were randomized into two groups: Reperfusion group: rats underwent coronary artery occlusion followed by reperfusion 2 hours after MI, a time too late to reduce infarct size. A control group: rats underwent permanent coronary artery occlusion followed by a sham operation 2 hours after MI. All rats received enalapril (2.0 +/- 0.2 mg/kg) daily in drinking water, started immediately after the second operation. Rats were sacrificed 2 weeks after coronary occlusion. Hearts were arrested and fixed at a constant pressure, then sectioned and photographed for morphometric analysis. RESULTS: Infarct size was similar in the reperfusion and control groups (23 +/- 2 vs 26 +/- 2%, p = NS). Septal thickness was also similar in both groups (1.8 +/- 0.1 vs 1.8 +/- 0.1 mm, p = NS). There was a trend towards thicker infarcts in the reperfusion group compared to the control group (0.84 +/- 0.06 vs 0.72 +/- 0.05 mm, p = 0.1). Compared to early ACE inhibition alone, late reperfusion combined with early ACE inhibition limited infarct expansion (expansion index, 1.13 +/- 0.12 vs 1.44 +/- 0.14, p < 0.05), prevented left ventricular (LV) dilation (LV volume, 0.30 +/- 0.02 vs 0.39 +/- 0.03 ml, p < 0.01) and prevented LV hypertrophy (LV weight, 0.71 +/- 0.18 vs 0.77 +/- 0.20 gm, p < 0.05). CONCLUSIONS: Late coronary artery reperfusion prevents infarct expansion, LV dilation and hypertrophy even when added to early ACE inhibitor therapy post MI. This suggests that late reperfusion may be beneficial in patients with acute MI treated with early ACE inhibitor therapy. PMID- 11504164 TI - Myocardial lactate release after intracoronary verapamil application in humans: acute effects of intracoronary verapamil on systemic and coronary hemodynamics, myocardial metabolism, and norepinephrine levels. AB - Coronary and systemic hemodynamic effects of verapamil have been investigated previously in detail. The acute impact of intracoronary verapamil on coronary hemodynamics has, however, not been correlated to simultaneously changes in myocardial metabolism or norepinephrine levels in humans. After bolus application of 1 mg verapamil into the left coronary artery of 52 patients scheduled for routine coronary angiography, heart rate (HR) remained unchanged, whereas mean arterial blood pressure (MAP) decreased (93.8 +/- 14.9 mmHg to 85.1 +/- 13.7 mmHg, p = 0.001). Coronary blood flow (CBF), calculated from intracoronary Doppler measurements and quantitative coronary angiography, increased after verapamil administration (28.5 +/- 16.7 ml/min to 66.2 +/- 41.8 ml/min, p < 0.001), whereas coronary vascular resistance index (CVRI) decreased (1.43 +/- 0.92 to 0.46 +/- 0.23, p < 0.001). Blood samples, taken simultaneously from the aorta (Ao) and coronary sinus (CS) at baseline and at maximal flow velocity, showed an increase in norepinephrine concentrations in Ao (209 +/- 151 ng/l to 283 +/- 195 ng/l, p < 0.001) and CS (233 +/- 162 ng/l to 323 +/- 248 ng/l, p = 0.004). Myocardial metabolism of pyruvate and free fatty acids were not affected. Glucose release was augmented and initial lactate consumption changed to a net lactate release into the CS (Ao to CS differences: glucose: -1.92 +/- 9.9 mg/dl to -12.8 +/- 22.8 mg/dl, p < 0.001; lactate: 0.07 +/- 0.2 mmol/l to -0.08 +/- 0.3 mmol/l, p = 0.001). Similar results were obtained for the extraction ratios and flux of these metabolites. There was a weak correlation between the increase in CBF and lactate release into the CS. This is the first report of unexpected myocardial lactate release following intracoronary verapamil administration in humans. This lactate release was paralleled by an increased glucose release into the CS at an unchanged metabolism of free fatty acids and pyruvate. One explanation for this unexplained lactate release during increased coronary blood flow might be a wash out phenomenon of lactate from previous ischemic areas, other explanations might be the induction of paradox myocardial ischemia and/or a steal effect. Further studies are necessary to explain these unexpected findings of increased coronary flow and myocardial lactate release. Until reliable explanations are pending, studies using only lactate release as a marker of myocardial ischemia, without taken coronary and systemic hemodynamic parameters into account, should be interpreted with caution. PMID- 11504165 TI - Comparison of the effect of perindopril and acebutolol on cerebral hemodynamics in hypertensive patients. AB - PURPOSE: The aim of the study was to compare effect of perindopril (4 mg once a day) versus acebutolol (400 mg once a day) on cerebral hemodynamics in hypertensive patients. METHODS: Thirty untreated patients with mild-to-moderate hypertension were studied. Drug influences on Doppler-derived parameters from the cerebral blood flow (CBF) velocity wave form were evaluated in a randomized, double blind, cross-over, placebo controlled study. The mean CBF velocity, pulsatility index (PI), cerebrovascular resistance and index of CBF were calculated from concomitant transcranial Doppler measurements and systemic blood pressure. RESULTS: Acebutolol and perindopril significantly decreased systolic, diastolic and mean blood pressure in relation to placebo. The mean value of CBF velocity increased to a comparable level after both drugs (54.9 +/- 9.1 cm/s on placebo vs 62.8 +/- 14.5 cm/s on perindopril p<0.01 and 61.4 +/- 9.2 cm/s on acebutolol, p<0.01). Also, the cerebrovascular resistance index decreased similarly after both drugs (2.26 +/- 0.35 on placebo vs 1.68 +/- 0.42 on perindopril p<0.01 and 1.7 +/- 0.36 on acebutolol p<0.01). The calculated CBF index increased significantly after each drug (25.23 +/- 7.99 on placebo vs 33.98 +/- 11.23 p<0.01 on perindopril and 30.90 +/- 8.04 on acebutolol p<0.01). However, perindopril augmented the CBF index more effectively than acebutolol (p<0.05). CONCLUSIONS: Among patients with mild-to-moderate hypertension both acebutolol and perindopril beneficially decreased cerebrovascular resistance and increased the CBF index in comparison with placebo. The increase of CBF index was greater after perindopril than acebutolol, which suggests a more significant improvement in cerebral perfusion by perindopril. The non-invasive transcranial Doppler ultrasonography method of CBF velocity measurement may contribute to choosing optimal antihypertensive therapies and to monitor their effect. PMID- 11504166 TI - The Carvedilol and ACE-Inhibitor Remodelling Mild Heart Failure EvaluatioN trial (CARMEN)--rationale and design. AB - Inhibition or reversal of ventricular remodelling in heart failure patients is regarded as of prime importance in the treatment of heart failure and in determining long term outcome. Recent studies have demonstrated that the addition of carvedilol to Angiotensin Converting Enzyme (ACE) inhibitors and other routine heart failure therapy results in a valuable improvement in the clinical status and life expectancy of mild, moderate and severe heart failure patients. ACE inhibitors have become the cornerstone of heart failure therapy. Also, carvedilol in combination with standard therapy (including ACE inhibitors) has demonstrable beneficial effects on left ventricular remodelling. Each new treatment has to be added, this quickly leads to polypharmacy, which may not be necessary and even unwanted in the individual patient, as the pharmacological profile of carvedilol compares favourably to ACE inhibitors, this suggests that it could challenge ACE inhibitors as first-line treatment for heart failure. The CARMEN trial (Carvedilol and ACE-Inhibitor Remodelling Mild Heart Failure EvaluatioN) was designed to compare the effects of carvedilol alone and of carvedilol plus an ACE inhibitor (enalapril) with the effect of an ACE inhibitor alone on different parameters of left ventricular remodelling as well as morbidity and mortality in patients with chronic mild heart failure, thereby allowing conclusions on whether combination therapy may be replaced by the multiple action adrenergic inhibitor carvedilol in the future. PMID- 11504167 TI - The EPHESUS trial: eplerenone in patients with heart failure due to systolic dysfunction complicating acute myocardial infarction. Eplerenone Post-AMI Heart Failure Efficacy and Survival Study. PMID- 11504168 TI - Opioid preconditioning extends to post-ischemic recovery of contractile function as shown with a novel opioid delta selective agonist, SB 235863A, that acts on the mitochondrial K(ATP) channel. PMID- 11504169 TI - Hormones, drugs, remodeling and outcome in heart failure and after myocardial infarction. PMID- 11504170 TI - Effect of angiotensin converting enzyme inhibitor on plasma von Willebrand factor level in patients with coronary artery disease. PMID- 11504171 TI - Scientific foundations of hormesis. Introduction. PMID- 11504172 TI - Hormesis: a generalizable and unifying hypothesis. AB - The present article represents a comprehensive effort to assess the hypothesis that hormesis is a highly generalizable biological phenomenon independent of environmental stressor, biological endpoint, and experimental model system. The evaluative methodology and complementary approaches employed to assess this question are (1) evolutionary biology-based theoretical paradigm; (2) evaluation of > 20,000 toxicology articles using a priori entry and evaluative criteria; (3) evaluation of 17 large-scale studies each providing data on numerous agents tested in the same experimental model by the same research team; (4) the assimilation of experimental pharmacological data on 24 receptor systems in which biphasic dose responses have been established reproducibly along with hormetic mechanism elucidation; and (5) assessment of the original hormesis database with 1600 dose-response relationships demonstrating evidence consistent with the hormesis hypothesis. The complementary approaches for assessing hormesis provided strong support for its credibility as a central biological theory based on its high frequency of occurrence and quantitative features of expression within microbe, plant, and invertebrate and vertebrate animal systems. The findings suggest that hormetic effects represent evolutionary-based adaptive responses to environmentally induced disruptions in homeostasis. Such adaptive responses, which are incorporated into organismal integrative physiological systems and now clarified at the mechanistic level for more than two dozen receptor systems, provide a cogent basis for the application of hormetic mechanisms in the elucidation of fundamental evolutionary-based biological processes and in the development of novel clinical modalities. PMID- 11504173 TI - Overcompensation stimulation: a mechanism for hormetic effects. AB - Whether hormetic responses result from a direct or an overcompensation type of stimulatory response has been an unresolved and contentious issue in both radiation and chemical toxicology. The goal of the present article is to identify numerous examples of overcompensation stimulation in the biological/biomedical literature and to evaluate their descriptive and quantitative features. The findings provide support for the hypothesis that hormetic dose-response relationships from a broad array of biological models can occur after an initial disruption in homeostasis. The finding also demonstrates the significant role of temporal factors in the assessment of dose response relationships. PMID- 11504174 TI - Agonist concentration gradients as a generalizable regulatory implementation strategy. AB - This article proposes a general mechanism by which biphasic dose-response relationships occur in pharmacological and toxicological experimental systems. Such biphasic responses are mediated via a strategy of differential binding to stimulatory and inhibitory receptor subtypes based on agonist concentrations. Such a strategy is widely seen in pharmacological systems and has been demonstrated in toxicological-related biphasic dose responses in which the treatment altered levels of endogenous agonists. PMID- 11504175 TI - Prostaglandins: biphasic dose responses. AB - Biphasic dose responses induced by prostaglandins are documented and assessed with respect to quantitative features of their dose-response functions, mechanistic foundation, and biological generalizability. Biphasic responses were seen for multiple endpoints, including DNA synthesis, neutrophil migration, corticosterone production, fibroblast proliferation, and other parameters. In addition, numerous nonsteroidal antiinflammatory agents and other drugs alter prostaglandin concentrations and/or transport in a biphasic dose- response manner. These findings have considerable implications for the understanding of basic biological regulatory processes as well as clinical practices. PMID- 11504176 TI - Nitric oxide: biphasic dose responses. AB - The capacity of nitric oxide (NO) to affect biphasic dose responses in pharmacological and toxicological systems was assessed. Numerous examples of such biphasic responses were documented, including osteoclast differentiation, various vascular responses, neutrophil migration, superoxide anion formation, exploratory behavior in rodents, vitamin D3 levels in macrophages, human sperm motility and mobility, myocardial contraction, and other functions. The quantitative features of the dose response indicated a maximum stimulatory response usually less than twofold greater than the controls. While the stimulatory range was variable, ranging from approximately 2.5 to 500-fold, the majority was < or = 10-fold. These findings indicate that biphasic dose-response relationships are common manifestations of the NO-induced effects. PMID- 11504177 TI - Estrogen and related compounds: biphasic dose responses. AB - This article documents and quantitatively assesses the capacity of estrogen, phytoestrogens, and antiestrogens to affect biphasic dose-response relationships in animal/human models and across a broad range of cell types, affecting multiple endpoints. The range of endpoints displaying such biphasic dose responses includes plasminogen activation, oxytocin secretion, angiogenesis, cell proliferation, bone growth, monocyte chemotaxis, secretion of various cytokines, and other effects. The quantitative features of the dose response relationships revealed that the magnitude of the stimulatory responses was typically less than twofold, whereas the stimulatory responses were markedly variable ranging from about 5- to 10(6)-fold. Mechanistic explanations of the biphasic responses are addressed. PMID- 11504178 TI - Androgens: biphasic dose responses. AB - Testosterone was reported to affect a variety of reproductive endpoints. More notable were the effects of dihydrotestosterone on cell proliferation in the prostate cancer cell model LNCaP and Sertoli cell function. Testosterone production was also biphasically affected by prolactin that was administered to adult testicular cells in vitro. PMID- 11504179 TI - Adrenergic receptors: biphasic dose responses. AB - Agonists and antagonists of the adrenergic receptor system were assessed for their capacity to affect dose-response relationships across biological model, tissue and endpoint. U shaped dose responses were commonly reported, affecting multiple endpoints such as smooth muscle contraction/relaxation, memory, blood pressure, sexual behavior, platelet aggregation, and others. In six of the endpoints studied, mechanistic foundations of the biphasic nature of the dose response were established. The quantitative features of the dose-response relationships indicated that the maximum stimulatory responses, with but one exception, were less than twofold greater than the controls. The range of stimulatory responses was generally within 10- to 1000-fold, except for the platelet aggregation endpoint where the range was 10(3) to 10(5). PMID- 11504180 TI - Adenosine: biphasic dose responses. AB - This article characterizes the occurrence of biphasic dose responses by adenosine and its stable analogs in numerous systems, including the respiratory tract, kidney, cardiovascular system, and brain. Considerable mechanistic research on various systems explored and clarified the interactions of the A-1 and A-2 receptors and their interactive influence on the occurrence of the biphasic dose responses. In general, the maximum stimulatory response was 2.5-fold or less than controls, while the stimulatory concentration range was quite variable ranging from 10- to 10(5)-fold. PMID- 11504181 TI - 5-Hydroxytryptamine (serotonin): biphasic dose responses. AB - This article briefly summarizes the occurrence of biphasic dose-response relationships associated with 5-hydroxytryptamine (5-HT) receptor systems, as well as 5-HT agonists and antagonists. Such biphasic responses have been reliably reported in multiple experimental settings that explored a broad range of responses, including aggressive behavior, cardiovascular functioning, muscle relaxation, and interactions with other receptor systems such as adrenergic interaction as well as with neurotransmitters such as NMDA. PMID- 11504182 TI - Dopamine: biphasic dose responses. AB - The present article indicates that dopamine and/or its agonists induce biphasic dose-response relationships for numerous endpoints. These include locomotion, pain sensitivity, blood pressure, prolactin secretion, oxytocin release, heart rate, memory, and neuronal adenylate cyclase activity. Biphasic responses were reported predominantly with male Sprague-Dawley rats, but also with mice, dogs, monkeys, and humans. Regardless of the model or endpoint the maximum changes from the control were always modest being within the 10 to 80% range. The range of stimulatory responses was quite variable, extending from slightly greater than a factor of 10 for the endpoints such as memory, pain-vocalization, and diastolic blood pressure to the 10(6) range for prolactin release and the 10(8) range for oxytocin release. Mechanistic studies suggested that the stimulatory and inhibitory effects of dopamine are mediated by different receptors or receptor subtypes having opposite actions and different ligand affinities. PMID- 11504183 TI - Opiates: biphasic dose responses. AB - It was shown that biphasic responses are commonly reported for opiates with respect to a broad range of animal models and endpoints. These endpoints include such diverse functions as blood pressure, muscle tension, breathing rates, hCG production, HIV production, neutrophil migration, ACTH production, protein binding, and neuronal functioning. Quantitative features of the dose-response relationships indicated that the maximum stimulatory responses were < or = 3-fold greater than the controls with most being between 10 to 70% greater than the controls. In contrast to the striking similarity in the maximum stimulatory response, there was marked variation with respect to the dose range of the stimulatory responses that varied from 10(1) to 10(10). Mechanistic assessments were conducted for most biphasic dose-response relationships and are addressed in detail. PMID- 11504184 TI - Amyloid beta-peptide: biphasic dose responses. AB - This article summarizes recent findings indicating that amyloid beta-peptide displays neurotoxic and neurotrophic effects, depending on concentration. Mechanistic findings revealed that reactive oxygen species mediate both the toxic and neurotropic responses as a function of concentration with low doses being neutotrophic, while higher doses were toxic. The data reveal a potential biological function for amyloid beta-peptide within an optimal concentration zone. These findings suggest the critical role of dose in understanding disease causation and clinical therapeutics for Alzheimer's disease. PMID- 11504185 TI - Apoptosis: biphasic dose responses. AB - The features of apoptotic dose responses typically have been evaluated over a broad dose-response continuum to differentiate apoptotic and necrotic responses. The present article specifically addresses the dose-response relationship for apoptosis in detail. It was determined that a number of well-documented cases exist in which the dose-response relationship for apoptosis is biphasic. The underlying mechanisms and biomedical implications of this biphasic dose-response phenomenon are discussed. PMID- 11504186 TI - Cell migration/chemotaxis: biphasic dose responses. AB - The dose-dependent effects of chemoattractants on target cell migration such as tumor cells, fibroblasts, and especially neutrophils were assessed. In general, the dose-response characteristics were strikingly biphasic in nature and were independent of both the target tissue and chemoattractant agent. The assessment included evaluations of the quantitative features of the dose-response relationship, mechanistic foundations of the biphasic responsiveness and clinical applications. PMID- 11504187 TI - A critique of "the scientific foundations of hormesis". PMID- 11504188 TI - Another view of the scientific foundations of hormesis. PMID- 11504189 TI - The future of hormesis: where do we go from here? AB - Although the occurrence of U-shaped dose responses in toxicology (i.e., hormetic effects) have been known for more than a century, the concept of hormesis has long been marginalized under the belief that such observations could be explained by a combination of poor study designs and normal variability. However, recent efforts have established that numerous highly reliable studies demonstrating hormetic effects exist and that such findings appear to be highly generalizable across species, endpoint measures, and class of agents assessed. In light of such a long and complicated history, and its significant biomedical/toxicological implications, this article explores the unique challenges that the concept of hormesis confronts in both the experimental and institutional domains with respect to assessing its scientific foundations and validity and the impediments to its intstitutional acceptance and use with society. This perspective is then immediately critiqued by five authors in subsequent articles. PMID- 11504190 TI - Reflections on the future of hormesis. PMID- 11504191 TI - A possible path forward for hormesis. PMID- 11504192 TI - The future of hormesis: what is the clinical relevance to hormesis? PMID- 11504193 TI - Hormesis - a pharmaceutical industry perspective. PMID- 11504194 TI - Hormesis in biogerontology. PMID- 11504195 TI - Commentary on white paper the future of hormesis: where do we go from here? Edward J. Calabrese. PMID- 11504196 TI - Evaluating the evidence for hormesis: a statistical perspective. AB - It is possible to account for hormesis under current regulatory guidelines by invoking criteria for departure from default risk assessment procedures. However, past experience suggests that it will be difficult to amass enough evidence for hormesis in an individual case to permit departure from default procedures. Accordingly, hormesis is likely to be important in agency risk assessments only if guidelines are modified to incorporate hormesis as a default assumption. This could be appropriate if hormesis is determined to be a universal or near universal phenomenon. Although there is ample evidence that hormesis occurs in many specific situations, the overall prevalence of hormesis is very difficult to evaluate based on currently available data. The lack of a valid statistical test for hormesis is a major limitation when evaluating evidence for hormesis. The attempts at estimating the prevalence of hormesis reviewed herein did not adequately control for false positives and/or may have had inadequate power to detect hormesis. Some suggestions are made for constructing a database and analyzing the data therein that would provide more readily interpretable information on the prevalence of hormesis. PMID- 11504197 TI - Radiation hormesis: data and interpretations. AB - Although the biological effects of large doses of ionizing radiation are predominantly harmful, low-to-intermediate doses have been observed to enhance growth and survival, augment the immune response, and increase resistance to the mutagenic and clastogenic effects of further irradiation in plants, bacteria, insects, and mammals. The existence of these stimulatory, or "adaptive", responses implies that the dose-response relationships for genetic and carcinogenic effects of radiation may be similarly biphasic, or hormetic, in nature, a possibility with far-reaching implications for radiation protection. As yet, however, the extent to which such responses may actually reduce the risks attributable to low-level irradiation remains to be determined, pending further elucidation of the relevant dose-response relationships and the apparent lack of responsiveness in some individuals. Therefore, further research is needed to resolve this question. PMID- 11504198 TI - FMD: strategy for dealing with the remaining 'hot spots'. PMID- 11504199 TI - Risk of infectious diseases on farms restocking after FMD. PMID- 11504200 TI - Serological, colostral and milk responses of cows vaccinated with a single dose of a combined vaccine against rotavirus, coronavirus and Escherichia coli F5 (K99). AB - Twenty-five Ayrshire/Friesian cows were vaccinated once with a new combined vaccine against rotavirus, coronavirus and Escherichia coli F5 (K99) or given a saline placebo 31 days before the first expected calving date. Blood samples were taken from the cows at intervals from vaccination until seven days after calving and from their calves up to 28 days after birth, and colostrum and milk samples were collected from the cows at intervals for 28 days after calving. There was a significant increase in the mean specific antibody titre against all three antigens in the serum of the vaccinated animals (even in the presence of pre existing antibody) which was accompanied by increased levels of protective antibodies to rotavirus, coronavirus and E coli F5 (K99) in their colostrum and milk for at least 28 days. PMID- 11504201 TI - Ultrasonography of the teat canal in cows and sheep. AB - When the isolated teat of a cow was examined with an 8.5 MHz linear array transducer in a vertical plane, the teat canal appeared as a thin, white line, bordered on each side by parallel, thick, grey-black bands. In a horizontal plane a comparable image was obtained. In a sheep, images of comparable quality were obtained with a 12 MHz transducer. Histological studies of the tissues whose removal led to the disappearance of this characteristic ultrasonographic appearance showed that it was associated with the stratified keratinised squamous epithelium with distinct papillae. The content of keratin in the stratum corneum was apparently responsible for the bright zone; the stratum lucidum was not visible, and the surrounding dark, less echoic area was associated with the stratum granulosum. Doppler echography in live animals confirmed this designation. The outer layers of the teat wall were more echogenic. PMID- 11504202 TI - Management of hepatitis B virus infections in two gibbons and a western lowland gorilla in a zoological collection. AB - Since 1996, three primates newly arrived at London Zoo have been found to be infected with hepatitis B virus. The species involved were white-cheeked gibbons (Hylobates leucogenys leucogenys and Hylobates leucogenys siki) and a western lowland gorilla (Gorilla gorilla gorilla). The protocols for the practical management of these cases, including the immunisation of susceptible non-human primates and the staff with recombinant hepatitis B vaccine are described, and the origin and evolution of hepatitis B infection in primates are discussed. PMID- 11504203 TI - Clinical features of renal tubular dysplasia, a new hereditary disease in Japanese Black cattle. AB - A new hereditary disease characterised by renal failure, poor growth and long hooves in Japanese Black cattle (wagyu) has been recognised in a region of central Japan since 1990. The number of calves affected has increased gradually, with the incidence reaching 17 of 485 (3.51 per cent) in 1995. Almost all the calves were slightly undersized at birth, and repeatedly had diarrhoea during the neonatal period. They began to show signs of growth retardation with proportional body and elongation of the hooves from about two to five months of age, but they had an almost normal or only slightly decreased appetite. The concentrations of urea nitrogen, creatinine and inorganic phosphorus in serum were high, and the affected calves excreted diluted urine frequently. Among 25 cases, the urine of 21 contained occult blood, 24 contained protein and two contained glucose. In 29 calves observed for 30 to 130 days, the course of the disease varied; in 21 of them it remained unchanged, six became gradually worse and two became severely debilitated and died. The disease was diagnosed as renal tubular dysplasia by histopathological examination. PMID- 11504204 TI - Possible bitertanol and fuberidazole poisoning in young pheasants. PMID- 11504205 TI - Contagious equine metritis in Turkey: first isolation of Taylorella equigenitalis from mares. PMID- 11504206 TI - Thiazole orange-positive platelets in healthy and thrombocytopenic dogs. PMID- 11504207 TI - Influence of different levels of steer serum on production of fertilisable buffalo oocytes in vitro. PMID- 11504208 TI - Kidney transplants in cats. PMID- 11504209 TI - Kidney transplants in cats. PMID- 11504210 TI - FMD inquiry. PMID- 11504211 TI - Equine granulocytic ehrlichiosis in the UK. PMID- 11504212 TI - Mouth ulcers in horses. PMID- 11504213 TI - Warble fly survey results, 2000/01. PMID- 11504214 TI - Psoroptes species in alpacas. PMID- 11504215 TI - Review of the origin of BSE: scrapie agent cannot be excluded. PMID- 11504216 TI - Health protocol for translocation of free-ranging elk. AB - When considering an elk (Cervus elaphus) restoration program, wildlife managers must evaluate the positive and negative elements of translocation. We prepared this protocol to give an overview of health considerations associated with translocation of elk, with an emphasis on movement of free-ranging elk from western North America to the southeastern USA. We evaluated infectious agents and ectoparasites reported in elk from two perspectives. First, we made a qualitative estimate of the ability of the agent to be introduced and to become established. This was done using a selected set of epidemiologic factors. Second, if there was a good possibility that the organism could become established in the release area, the potential pathological consequences for elk and other wildlife, domestic animals, and humans were assessed via examination of the literature and consultation with other animal health specialists. The results of these evaluations were used to classify infectious agents and ectoparasites as low risk (n = 174), unknown risk (n = 10), and high risk (n = 9). We classified Anaplasma marginale, Anaplasma ovis, Mycobacterium paratuberculosis, Pasteurella multocida serotype 3, Elaphostrongylus cervi, Dicrocoelium dendriticum, Fascioloides magna, Echinococcus granulosus, Dermacentor albipictus, and Otobius megnini as unknown risks. High risk infectious agents and ectoparasites were the agent of chronic wasting disease, Brucella abortus, Mycobacterium bovis, Dermacentor andersoni, Ixodes pacificus, and Psoroptes sp. Parelaphostrongylus tennis, Elaeophora schneideri, and a Babesia sp. are parasites endemic in the southeastern USA that may present a "reverse risk" and adversely affect elk if released in some parts of the region. We developed a five-component protocol to reduce the risk of introduction of high risk infectious agents and ectoparasites that included: (1) evaluation of the health status of source populations, (2) quarantines, (3) physical examination and diagnostic testing, (4) restrictions on translocation of animals from certain geographic areas or populations, and (5) prophylactic treatment. PMID- 11504217 TI - Guidelines for the field evaluation of desert tortoise health and disease. AB - Field evaluation of free-ranging wildlife requires the systematic documentation of a variety of environmental conditions and individual parameters of health and disease, particularly in the case of rare or endangered species. In addition, defined criteria are needed for the humane salvage of ill or dying animals. The purpose of this paper is to describe, in detail, the preparation, procedures, and protocols we developed and tested for the field evaluation of wild desert tortoises (Gopherus agassizii). These guidelines describe: preparations for the field, including developing familiarity with tortoise behavior and ecology, and preparation of standardized data sheets; journal notes to document background data on weather conditions, temperature, rainfall, locality, and historic and recent human activities; procedures to prevent the spread of disease and parasites; data sheets for live tortoises to record tortoise identifiation, location, sex, body measurements and activity; health profile forms for documenting and grading physical abnormalities of tortoise posture and movements, general condition (e.g., lethargy, cachexia), external parasites, and clinical abnormalities associated with shell and upper respiratory diseases; permanent photographic records for the retrospective analysis of progression and regression of upper respiratory and eye diseases, analysis of shell lesions and evaluation of growth and age; and indications and methods for salvaging ill or dying tortoises for necropsy evaluation. These guidelines, tested on 5,000 to 20,000 tortoises over a 10 to 27 yr period, were designed to maximize acquisition of data for demographic, ecological, health and disease research projects; to reduce handling and stress of individual animals; to avoid spread of infectious disease; to promote high quality and consistent data sets; and to reduce the duration and number of field trips. The field methods are adapted for desert tortoise life cycle, behavior, anatomy, physiology, and pertinent disease; however the model is applicable to other species of reptiles. Comprehesive databases of clinical signs of disease and health are crucial to research endeavors and essential to decisions on captive release, epidemiology of disease, translocation of wild tortoises, breeding programs, and euthanasia. PMID- 11504218 TI - Health effects following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards. AB - Permanent approval of shot composed of tungsten-iron and tungsten-polymer for waterfowl hunting by the U.S. Fish and Wildlife Service was pending the results of the present study that examined the health and reproductive effects of the two shot types on mallards (Anas platyrhynchos) over a 150-day period. We collected data pertaining to the effects of tungsten-iron and tungsten-polymer shot on mortality, body weight, organ weight, tissue pathology, and shot erosion. Thirty two bird groups (sexes equal) of adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten-polymer shot on days 0, 30, 60, 90, and 120 of a 150-day trial (26 January 1998 to 25 June 1998). An additional 12 mallards (sexes equal) were dosed orally with eight #4 lead shot (positive control) on day 0 of the study. All lead-dosed ducks died by day 25, whereas no ducks died in the other treatment groups. Significant liver hemosiderosis was present in all control and tungsten-iron-dosed males, in five of eight control and three of eight tungsten-iron-dosed females, and in one tungsten-polymer-dosed male examined. The rate of shot erosion was highest for tungsten-polymer shot (99%), followed by tungsten-iron (72%), and steel (55%) shot. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards did not have deleterious health effects during the 150-day trial based on mortality, body weights, organ weights, and histology of the liver and kidneys. PMID- 11504219 TI - Hematological effects and metal residue concentrations following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards. AB - The U.S. Fish and Wildlife Service required a chronic dosing study that assessed the health and reproductive effects of tungsten-iron and tungsten-polymer shot in adult game-farm mallards (Anas platyrhynchos) prior to granting permanent approval of the shot for waterfowl hunting. Herein, we present the effects of tungsten-iron and tungsten-polymer shot on various hematologic parameters and metal residue concentrations in the femur, liver, kidneys, and gonads. Thirty-two bird groups (sexes equal) of adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten-polymer shot on days 0, 30, 60, 90, and 120 of a 150 day trial (26 January 1998 to 25 June 1998). An additional 12 mallards (sexes equal) received eight #4 lead shot (positive control) on day 0 of the study. Lead-dosed mallards had significantly decreased hematocrit, hemoglobin concentration, and whole-blood delta aminolevulinic acid dehydratase activity on day 7, as well as significant changes in a number of plasma chemistry parameters compared to ducks in the control, tungsten-iron, or tungsten-polymer groups. Mallards dosed with tungsten-iron or tungsten-polymer shot had occasional significant differences in hematocrit and plasma chemistry values when compared to control mallards over the 150 day period, but these changes were not considered to be indicative of deleterious effects. Low concentrations of tungsten were detected in gonad and kidney samples from males and females and in liver samples from females dosed with tungsten-polymer shot. Tungsten was also detected in femur samples from tungsten-polymer-dosed mallards. Higher concentrations of tungsten were detected in femur, liver, kidney, and gonad samples from tungsten-iron-dosed ducks. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards did not cause adverse hematological effects during the 150 day trial. Concentrations of tungsten in the femur, liver, kidneys, and gonads were generally higher in tungsten-iron-dosed ducks when compared to tungsten-polymer-dosed ducks. PMID- 11504220 TI - Reproductive effects and duckling survivability following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards. AB - Tungsten-iron and tungsten-polymer shot were given conditional approval for waterfowl hunting by the U.S. Fish and Wildlife Service based partly on the results of a 30-day acute toxicity trial utilizing mallards (Anas platyrhynchos). Final approval of the two tungsten-containing shot was contingent on the results of a 150-day study that assessed the health and reproductive effects of tungsten iron and tungsten-polymer shot in adult mallards. Reproductive data are presented in this paper. Sixteen male and 16 female adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten polymer shot on days 0, 30, 60, 90, and 120 of a 150-day trial (26 January 1998 to 25 June 1998). Reproductive performance was assessed during the last 90 days (day 61 to day 150) of the trial. There were no significant differences in egg production and fertility and hatchability of eggs from tungsten-iron- and tungsten-polymer-dosed ducks compared to control ducks. There was no evidence of differences in percent survivability and body weight of ducklings from tungsten iron and tungsten-polymer mallards compared to ducklings from control ducks. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards during the 150 day trial did not adversely affect reproduction or their offspring. PMID- 11504221 TI - Aflatoxin production in supplemental feeders provided for northern bobwhite in Texas and Oklahoma. AB - Mycotoxins are toxic metabolites produced by varous species of fungi. Aflatoxin (AF), a particular type of mycotoxin, can negatively impact many wildlife species in the laboratory; however, the magnitude of the problem in the field environment is unclear. Wild birds generally consume a combination of native foods and agricultural grains. A common practice in which birds, such as northern bobwhite (Colinus virginianus), contact stored agricultural grain is through supplemental feeding. This feeding practice may promote the production of AF. The objectives of this study were to (1) examine AF production in supplemental feeders and (2) examine the relationship between weather and AF production in supplemental feeders. Samples were collected from supplemental feeders from November through February of 1996-97 and 1997-98. Mean monthly AF concentration of samples from feeders ranged from 0.57+/-2.86 to 15.47+/-14.69 ppb. Aflatoxin concentration in supplemental feeders increased from pre-sample to one month after filling the feeders each year. AF production in supplemental feeders was highly variable among months with no real temporal pattern between years. Instead, AF production was related to the highly variable relative humidity of the study area which influences moisture content of grain. Average relative humidity can be used to predict AF production. PMID- 11504222 TI - Hemograms for and nutritional condition of migrant bald eagles tested for exposure to lead. AB - Plasma proteins, hematocrit, differential blood counts were examined and nutritional condition was estimated for bald eagles (Haliaeetus leucocephalus) trapped (n = 66) during antumn migration, 1994-95 at Galloway Bay (Saskatchewan, Canada), for the purposes of estimating prevalence of exposure to lead. Sex and age differences in hematocrit and plasma proteins were not observed; however, female eagles exhibited larger median absolute heterophil counts than males. Hematologic values were similar to those previously reported from eagles in captivity. Departures from expected hematological values from a healthy population of eagles were not observed in birds with elevated levels of blood lead (> or =0.200 microg/ml). Similarly, nutritional condition was not related to blood-lead concentrations. Therefore, it appears that lead exposure in this population was below a threshold required to indicate toxicological alteration in the hematological values and index of nutritional condition that we measured. PMID- 11504223 TI - Biomarker responses in river otters experimentally exposed to oil contamination. AB - Investigations in Prince William Sound (Alaska, USA) following the Exxon Valdez oil spill (EVOS) revealed that river otters (Lontra canadensis) on oiled shores had lower body mass and elevated values of biomarkers, than did otters living on nonoiled shores. In addition, otters from oiled areas selected different habitats, had larger home ranges, and less diverse diets than animals living in nonoiled areas. These differences between river otters from oiled shores and those from nonoiled areas strongly suggested that oil contamination had an effect on physiological and behavioral responses of otters. In this study, we explored the effects of crude oil contamination on river otters experimentally. We hypothesized that exposure to oil would result in elevated values of biomarkers, indicating induced physiological stress. Fifteen wild-caught male river otters were exposed to two levels of weathered crude oil (i.e., control, 5 ppm/day/kg body mass, and 50 ppm/day/kg body mass) under controlled conditions in captivity at the Alaska Sealife Center in Seward (Alaska, USA). Responses of captive river otters to oil ingestion provided mixed results in relation to our hypotheses. Although hemoglobin (Hb, and associated red blood cells) and white blood cells, and possibly interleukin-6 immunoreactive responded in the expected manner, other parameters did not. Aspartate aminotransferase, alanine aminotransferase, and haptoglobin (Hp), did not increase in response to oiling or decreased during rehabilitation. Conversely, principle-component analysis identified values of alkaline phosphatase as responding to oil ingestion in river otters. Our results suggested that opposing processes were concurring in the oiled otters. Elevated production of Hp in response to tissue damage by hydrocarbons likely occurred at the same time with increased removal of Hp-Hb complex from the serum, producing an undetermined pattern in the secretion of Hp. Thus, the use of individual biomarkers as indicators of exposure to pollutants may lead to erroneous conclusions because interactions in vivo can be complicated and act in opposite directions. Additionally, the biomarkers used in investigating effects of oiling on live animals usually are related to the heme molecule. Because of the opposing processes that may occur within an animal, data from a suite of heme-related biomarkers may produce results that are difficult to interpret. Therefore, we advocate the exploration and development of other biomarkers that will be independent from the heme cycle and provide additional information to the effect of oiling on live mammals. PMID- 11504224 TI - Mineral and heavy metal status as related to a mortality event and poor recruitment in a moose population in Alaska. AB - Moose (Alces alces) found dead (FD) and hunter-killed (HK) in 1995 on the north slope of Alaska (USA) in the Colville River Drainage were evaluated for heavy metal and mineral status. Compared to previous reports for moose and domestic cattle, and data presented here from Alaska moose outside the Colville River area, levels of Cu were determined to be low in hoof, hair, liver, kidney, rumen contents, and muscle for these north slope moose. Iron (Fe) was low in muscle as well. These findings, in conjunction with evidence of poor calf survival and adult mortality prompted investigation of a mineral deficiency in moose (serum, blood, and hair) captured in the spring of 1996 and 1997. Captured males had higher Ca, Zn and Cu levels in hair than captured females. Female moose hair samples were determined to be low (deficient) in Cu, Ca, Fe, and Se with mean levels (ppm) of 2.77, 599.7, 37.4, and 0.30, respectively. Serum Cu level was low, and to a lesser degree Zn was deficient as well. Whole blood (1997 only) was marginally deficient in Se and all animals were deficient in Cu. Based on whole blood, sera and hair, Cu levels were considered low for moose captured in spring 1996 and 1997 in the Colville River area as compared to published data and other populations evaluated in this study. Low levels of ceruloplasmin activity support this Cu deficiency theory. Evidence indicates that these moose are deficient in Cu and other minerals; however, the remote location precluded sufficient examination of animals to associate this apparent deficiency with direct effects or lesions. Renal levels of Cd increased with age at expected levels. PMID- 11504225 TI - Brucella sp. antibodies in polar bears from Svalbard and the Barents Sea. AB - A prevalence of 5.4% of anti-Brucella sp. antibodies was found in plasma samples from 297 polar bears (Ursus maritimus) from Svalbard and the Barents Sea. Plasma was tested by the classical brucellosis tests Slow Agglutination of Wright (SAW), EDTA modified SAW and Rose Bengal test, as well as by an indirect Protein A ELISA. Only samples classified as positive in all tests were regarded as containing anti-Brucella sp. antibodies. A significant west to east increase in the proportion of bears with anti-Brucella sp. antibodies was found, with 3.6% (n = 253) at Svalbard (Spitsbergen, Nordaustlandet, Edgeoya, Barentsoya and Hopen), and 15.9% (n = 44) in the central Barents Sea. Anti-Brucella sp. antibodies were previously found in ringed seals (Phoca hispida) and harp seals (Phoca groenlandica) from the same geographical areas. The ringed seal is an important prey species for the Svalbard polar bear population, and may thus be a source of brucellosis for the bears. There are no indications of reproductive disorders caused by Brucella sp. or other infectious agents in our study polar bear population. Potential impacts of Brucella sp. exposure on individuals or the population are unknown. PMID- 11504226 TI - Experimental infection of nontarget species of rodents and birds with Brucella abortus strain RB51 vaccine. AB - The Brucella abortus vaccine strain RB51 (SRB51) is being considered for use in the management of bnucellosis in wild bison (Bison bison) and elk (Cervus elaphus) populations in the Greater Yellowstone Area (USA). Evaluation of the vaccines safety in non-target species was considered necessary prior to field use. Between June 1998 and December 1999, ground squirrels (Spermophilus richardsonii, n = 21), deer mice (Peromyscus maniculatus, n = 14), prairie voles (Microtus ochrogaster, n = 21), and ravens (Corvus corax, n = 13) were orally inoculated with SRB51 or physiologic saline. Oral and rectal swabs and blood samples were collected for bacteriologic evaluation. Rodents were necropsied at 8 to 10 wk and 12 to 21 wk post inoculation (PI), and ravens at 7 and 11 wk PI. Spleen, liver and reproductive tissues were collected for bacteriologic and histopathologic evaluation. No differences in clinical signs, appetite, weight loss or gain, or activity were observed between saline- and SRB51-inoculated animals in all four species. Oral and rectal swabs from all species were negative throughout the study. In tissues obtained from SRB51-inoculated animals, the organism was isolated from six of seven (86%) ground squirrels, one of six (17%) deer mice, none of seven voles, and one of five (20%) ravens necropsied at 8, 8, 10, and 7 wk PI, respectively. Tissues from four of seven (57%) SRB51-inoculated ground squirrels were culture positive for the organism 12 wk PI; SRB51 was not recovered from deer mice, voles. or ravens necropsied 12, 21, or 11 wk, respectively, PI. SRB51 was not recovered from saline-inoculated ground squirrels, deer mice, or voles at any time but was recovered from one saline inoculated raven at necropsy, 7 wk PI, likely attributable to contact with SRB51 inoculated ravens in an adjacent aviary room. Spleen was time primary tissue site of colonization in ground squirrels, followed by the liver and reproductive organs. The results indicate oral exposure to SRB51 does not produce morbidity or mortality in ravens, ground squirrels, deer mice, or prairie voles. PMID- 11504227 TI - Persistent Ehrlichia chaffeensis infection in white-tailed deer. AB - Four white-tailed deer (Odocoileus virginianus) were inoculated intravenously with a deer-origin isolate (15B-WTD-GA) of Ehrlichia chaffeensis. The course of infection was monitored using indirect fluorescent antibody (IFA), polymerase chain reaction (PCR), and culture over a 9 m period. All deer became rickettsemic within 24 days post inoculation (DPI), and all developed antibody titers >1:64 to E. chaffeensis by 17 DPI. Titers in all deer fell below 1:64 during 87 to 143 DPI. One deer exhibited a second period of seropositivity (peak titer of 1:256) from 207 to 271 DPI but was culture and PCR negative during this period. Rickettsemia was confirmed by reisolation of E. chaffeensis as late as 73 to 108 DPI in three deer. Positive PCR results were obtained from femur bone marrow of one deer and from rumenal lymph node of another (leer at 278 DPI. None of the deer developed clinical signs, hematologic abnormalities, or gross or microscopic lesions attributable to E. chaffeensis. Two uninoculated control deer were negative on all tests through 90 DPI at which time they were removed from the study. Herein we confirm that white-tailed deer become persistently infected with E. chaffeensis, have initial rickettsemias of several weeks duration and may experience recrudescence of rickettsemia, which reaffirm the importance of deer in the epidemiology of E. chaffeensis. PMID- 11504228 TI - Mycoplasmosis in captive crows and robins from Minnesota. AB - Mycoplasma sturni is a recently described organism previously associated with conjunctivitis in European starlings (Sturnus vulgaris), northern mockingbirds (Mimus polyglottos) and blue jays (Cyanocitta cristata). Herein we describe the isolation of M. sturni from an American crow (Corvus brachyrhynchos) presenting with conjunctivitis. A nested-PCR was designed for identification of M. sturni in clinical specimens and the sensitivity of the reaction was found to be 10 colony changing units. The organism was found in asymptomatic American crows caged with a nestmate of the crow with conjunctivitis. Mycoplasma sturni also was found in asymptomatic American robins (Turdus migratorius) and in a European starling (Sturnus vulgaris) housed at the same facility as the crows. Heterogenity of M. sturni isolates from different host species was found by random amplified polymorphic DNA (RAPD) analyses. Heterogeneity also was found among M. sturni isolates recovered from American crows. We suggest that M. sturni can successfully infect American crows and American robins with or without the presence of clinical disease. Furthermore, we demonstrate that nested-PCR is an effective method for the detection of M. sturni and that substantial genetic heterogeneity exists among natural isolates of this bacterial pathogen. PMID- 11504229 TI - Altered predation susceptibility of mosquitofish infected with Eustrongylides ignotus. AB - Eustrongylides ignotus is a parasitic nematode whose definitive hosts are often piscivorous wading birds (Ciconiiformes). Several species of small fishes are intermediate hosts, while larger predatory fish may be paratenic (transport) hosts. We examined predation susceptibility of infected mosquitofish (Gambusia holbrooki) to three species of predatory fishes, including juvenile largemouth bass (Micropterus salminoides), warmouth (Lepomis gulosus), and bluegill (Lepomis macrochirus). A 250 L aquarium with removable plexiglass divider and remote observation windows was constructed. Aquatic macrophytes were placed in the tank to provide refuge for the fishes. Predatory fish were allowed to acclimate to one half of the tank, while one infected and one uninfected mosquitofish were placed in the other. The divider was removed and an observer recorded the number of capture attempts and time required for capture. Predators were observed for behavioral alterations for 4 days after ingestion of infected mosquitofish, then examined at necropsy. Infected prey were selected preferentially in 31 of 38 (82%) trials. The number of capture attempts was 2.7+/-0.2 (x +/- SE) for infected fish and 3.9+/-0.4 for uninfected fish. Mean time of capture was 12.4+/ 1.6 min for infected fish and 21.7+/-2.9 for uninfected fish. Because of these differences, infected mosquitofish were more susceptible to predation (P < 0.01) than uninfected fish. Aberrant behavior including lethargy, convulsions, and buoyancy abnormalities was observed in eight (67%) predatory fish. At necropsy, larvae of E. ignotus were found in the coelomic cavity, viscera, and swim bladders of predators. Parasite-induced behavior modification of intermediate hosts may predispose them to predation by wading birds and thereby facilitate the transmission of this nematode in natural populations. PMID- 11504230 TI - Reversible immobilization of eurasian otters with a combination of ketamine and medetomidine. AB - The efficacy and safety of the combination of medetomidine and ketamine was examined in order to establish an adequate chemical immobilization protocol in the Eurasian otter (Lutra lutra) for use during translocation projects in Spain. Thirty-eight Eurasian otters ranging in body mass from 3 to 8.7 kg (mean 5.3 kg) were successfully anesthetized on 82 occasions. The dosage of ketamine was 5.1+/ 0.8 (3.4-6.6) mg/kg (mean +/- SD; range) combined with medetomidine at a dosage of 51+/-8 Rg/kg (34-66 microg/kg). In most cases anaesthetic effect occurred within 3 min and the mean induction time was 5.5+/-3.2 min. The mean pulse rate was 95 beats/min. The mean respiratory rate was 32 respirations/min while the relative oxyhemoglobin saturation was 93%. According to these results, this anesthetic protocol is considered safe and can be recommended in wild caught Eurasian otters for immobilization during translocation projects. It is safe, rapid and can be reversed when needed with atipamezole. However caution is required as heart depression resulting in bradychardia may occur. PMID- 11504231 TI - Use of medetomidine-zolazepam-tiletamine with and without atipamezole reversal to immobilize captive California sea lions. AB - From June 1998 to August 1999, 39 California sea lions (Zalophus californianus) were immobilized at a rehabilitation center in northern California (USA) using medetomidine plus zolazepam and tiletamine (MZT), alone and in combination with isoflurane, with atipamezole reversal. Animals were given 70 microg/kg medetomidine with 1 mg/kg of a 1:1 solution of tiletamine and zolazepam intramuscularly. Mean (+/-SD) time to maximal effect was 5+/-3 min. At the end of the procedure, animals were given 200 microg/kg atipamezole intramuscularly. Immobilization and recovery times were, respectively, 28+/-18 and 9+/-7 min for 15 animals maintained with MZT alone and 56+/-47 and 9+/-6 min for 18 animals intubated and maintained with isoflurane. One mortality occurred during anesthesia. Other disadvantages of the MZT combination included some prolonged ataxia, weakness and disorientation during recovery. However, the use of MZT resulted in faster induction and a more reliable plane of anesthesia that was reversible with atipamezole and safer than other previously used intramuscular agents. Physiological parameters including heart rate, respiratory rate, temperature, pulse oximeter saturation, and end-tidal carbon dioxide were monitored. PMID- 11504232 TI - Immune status of free-ranging green turtles with fibropapillomatosis from Hawaii. AB - Cell-mediated and humoral immune status of free-ranging green turtles (Chelonia mydas) in Hawaii (USA) with and without fibropapillornatosis (FP) were assessed. Tumored and non-tumored turtles from Kaneohe Bay (KB) on the island of Oahu and from FP-free areas on the west (Kona/Kohala) coast of the island of Hawaii were sampled from April 1998 through February 1999. Turtles on Oahu were grouped (0-3) for severity of tumors with 0 for absence of tumors, 1 for light, 2 for moderate, and 3 for most severe. Turtles were weighed, straight carapace length measured and the regression slope of weight to straight carapace length compared between groups (KB0, KB1, KB2, KB3, Kona). Blood was assayed for differential white blood cell count, hematocrit, in vitro peripheral blood mononuclear cell (PBMC) proliferation in the presence of concanavalin A (ConA) and phytohaemagglutinin (PHA), and protein electrophoresis. On Oahu, heterophil/lymphocyte ratio increased while eosinophil/monocyte ratio decreased with increasing tumors score. Peripheral blood mononuclear cell proliferation indices for ConA and PHA were significantly lower for turtles with tumor scores 2 and 3. Tumor score 3 turtles (KB3) had significantly lower hematocrit, total protein, alpha 1, alpha 2, and gamma globulins than the other four groups. No significant differences in immune status were seen between non-tumored (or KB1) turtles from Oahu and Hawaii. There was no significant difference between groups in regression slopes of body condition to carapace length. We conclude that turtles with severe FP are imunosuppressed. Furthermore, the lack of significant difference in immune status between non-tumored (and KB1) turtles from Oahu and Kona/Kohala indicates that immunosuppression may not be a prerequisite for development of FP. PMID- 11504233 TI - Survey of bat populations from Mexico and Paraguay for rabies. AB - A mammalian survey was conducted in Mexico (October 1994-January 1996) and in Paraguay (August 1996-March 1997); a complete specimen was collected for each bat in the survey, including primary voucher specimen, ectoparasites, karyotype, and various frozen tissues. The surveys combined provided 937 brain samples (65 bat species) for rabies diagnosis. One male Lasiurus ega, collected in Paraguay, tested positive for the rabies virus (overall prevalence rate of 0.1%). Nucleotide sequence from a 300 bp region of the rabies nucleoprotein gene was compared with sequence obtained from representative rabies virus samples in the repository at the Centers for Disease Control and Prevention (Atlanta, Georgia, USA). Rabies virus extracted from the brain material of L. ega differed by only one nucleotide from a 300 bp consensus sequence (>99% homology) derived from samples for the variant of rabies virus transmitted by Lasiurus cinereus. Lasiurus ego differed by approximately 15% for the variant transmitted by Desmodus rotundus. Phylogenetic analysis found no evidence to suggest L. ego is a reservoir for rabies antigenic variant 6. The most likely explanation for rabies in L. ega was infection following contact with a rabid L. cinereus. PMID- 11504234 TI - Parvovirus infections in wild carnivores. AB - Various parvoviruses infect carnivores and can cause disease. In this review article the knowledge about infections of free-ranging or captive carnivores with the feline parvoviruses, feline panleukopenia virus, and canine parvovirus, including the antigenic types CPV-2a and -2b, as well as Aleutian disease of mink virus and minute virus of canines are summarized. Particular emphasis is placed on description of the evolution of canine parvovirus which apparently involved wild carnivore hosts. PMID- 11504235 TI - Tuberculous lesions in free-ranging white-tailed deer in Michigan. AB - Descriptions of the anatomical distribution of Mycobacterium bovis gross lesions in large samples of white-tailed deer (Odocoileus virginianus) are lacking in the scientific literature. This report describes the distribution of gross lesions in the 58 white-tailed deer that cultured positive for M. bovis among the 19,500 submitted for tuberculosis testing in Michigan (USA) in 1999. For the vast majority (19,348) of those tested, only the head was submitted; for others, only extracranial tissues (33) or both the head and extracranial tissues (119) were available. Among those deer that cultured positive, cranial gross lesions were noted most frequently in the medial retropharyngeal lymph nodes, although solitary, unilateral parotid lymph node lesions also were found. Extracranial lesions occurred most commonly in the thorax. The distribution of lesions largely agreed with the few existing case reports of the M. bovis in white-tailed deer, although gross lesions were also found in sites apparently not previously reported in this species (liver, spleen, rumen, mammary gland). Some practical issues that may assist future surveillance and public education efforts are also discussed. PMID- 11504236 TI - Granulocytic ehrlichiosis in a roe deer calf in Norway. AB - A case of granulocytic ehrlichiosis is described in a roe deer (Capreolus capreolus) calf from Norway. The calf was heavily infested with Ixodes ricinus and died from Escherichia coli septicemia. Granulocytic Ehrlichia sp. was detected by polymerase chain reaction (PCR) from several organs and sequence determination identified a variant of human granulocytic ehrlichiosis (HGE) agent. This is the first report of a possible clinical granulocytic Ehrlichia sp. infection in a roe deer. PMID- 11504237 TI - Enterotoxigenic Escherichia coli infection in captive black-footed ferrets. AB - Enterotoxigenic Escherichia coli with genes for heat stabile toxins Sta and STb was isolated from the gastrointestinal tract and multiple visceral organs of three adult and three juvenile black-footed ferrets (Mustela nigripes) that died in a captive breeding colony between 24 May 1998 and 2 July 1998. Similar isolates were obtained from rectal swabs of one adult and one juvenile that were clinically ill. All were fed a diet composed of mink chow, raw rabbit meat, beef liver powder, blood meal and lard. Escherichia coli of the same toxin genotype was isolated from the mixed ration. Clinical signs included sudden death, dehydration, anorexia and diarrhea. Necropsy lesions included acute enteritis with large numbers of rod shaped bacteria microscopically visible on intestinal villi. PMID- 11504238 TI - Safety of Brucella abortus strain RB51 in deer mice. AB - Brucella abortus strain RB51 is an approved brucellosis vaccine for use in cattle that may have potential as an oral vaccine for use in elk (Cervus elaphus) and/or bison (Bison bison). This study was designed to determine effects of strain RB51 on deer mice (Peromyscus maniculatus), a nontarget species that could have access to treated baits in a field situation. In February 1994, 90 mice were orally dosed or intraperitoneally injected with 1 x 10(8) colony forming units strain RB51 and 77 controls were similarly dosed with sterile saline. At weekly intervals through early April 1994, 4 to 6 mice from each group were euthanized, gross necropsies performed, spleens and uteruses cultured, and tissues examined histologically. All orally inoculated mice cleared the infection by 6 wk post inoculation (PI). While most of the injected mice cleared the infection by 7 wk PI, a few required 9 wk. There were minimal adverse effects attributable to strain RB51. Apparently, strain RB51 would not negatively impact P. maniculatus populations if it were used in a field situation. Also, deer mice appear to be able to clear the vaccine in 6 to 9 wk, thus the probability of these mice transmitting the vaccine to other animals is low. PMID- 11504239 TI - Otodectic otoacariasis in free-ranging Eurasian lynx in Sweden. AB - An infestation with Otodectes cynotis, the ear mite of cats and dogs, was observed in three free-ranging Eurasian lynx (Lynx lynx) killed in Sweden. The ear canals were obstructed by waxy secretions and exfoliated epithelium. Histologically, there were hyperkeratosis and acanthosis, and the epithelial surface was overlained by hyperkeratotic and parakeratotic crusts with mites, mite detritus and cerumen. In the subcutis there was a slight to moderate infiltration of lymphocytes and macrophages. The ceruminous glands were bypertrophic and hyperplastic, and there was also an hyperplasia of the sebaceous glands. The lesions seemed to correlate with the degree of infestation. To our knowledge, this is the first report of otoacariasis in free-ranging lynx. PMID- 11504240 TI - Notoedric mange in western gray squirrels from Washington. AB - From February 1998 to July 1999, 65 western gray squirrels (Sciurus griseus griseus) were trapped at three sites in Klickitat County, Washington (USA) as part of a home range and habitat use study. No squirrels (0/9) with mange lesions were identified in the initial trapping session (February and March 1998). During all subsequent trapping sessions (August 1998 through July 1999), squirrels with lesions consistent with notoedric mange, caused by the mite Notoedres centrifera (douglasi), were captured or recaptured at all three study sites. The diagnosis was confirmed by histopathology and examination of mites obtained from skin scrapings from two affected squirrels. Of the 56 squirrels captured from August 1998 to July 1999, 33 (59%) had characteristic mange lesions, and 14 (42%) affected squirrels died directly of mange or of secondary complications of mange. Only four breeding females of 22 radio-collared animals (males and females) in the study population were known to have survived the mange outbreak (12 died, 6 missing). Factors potentially contributing to this mange outbreak include a mast crop failure in the fall of 1998 and transmission of mites from animial to animal during trapping and processing sessions. PMID- 11504241 TI - Trombiculiasis in the Florida black bear. AB - We found trombiculid mite (Trombiculidae) infestations in 32 of 101 (32%) freeranging Florida black bears (Ursus americanus floridanus) live-captured or necropsied in Florida from January 1999 to April 2000. Prevalence of chigger infestation was greatest in June with no infestations seen October to March. Chigger infestations were recognized as accumulations of bright orange granular material usually associated with hair shafts. Mites were found in clusters of one to 102 (mean +/- SD = 8.5+/-19.5) and were distributed primarily over the ventral abdomen and thorax, inguinal and axillary regions, and proximal medial aspect of the extremities. Mites were identified as larval Eutrombicula splendens. Cutaneous lesions were seen in two of 32 (6%) infested bears. PMID- 11504242 TI - Susceptibility of common voles to experimental toxoplasmosis. AB - Common voles (Microtus arvalis) in groups of nine to 10 animals were inoculated per os with a dose of 1, 10, 1x10(2), 1x10(3), and of the K1 strain of Toxoplasma gondii. All the common voles inoculated with 1 to 1 x 10(3) oocysts remained subclinical and survived. Three of the 10 voles inoculated with 1 x 10(4) oocysts died between days 7 and 12 post inoculation (p.i.). Antibodies were demonstrated in all the infected voles killed on day 60 p.i. The highest antibody titres in voles detected by the dye test (DT) and latex agglutination test (LAT) were 1,024 and 1,280, respectively. PMID- 11504243 TI - Microscopic evidence of adenoviral infection in a muskrat in Illinois. AB - A wild muskrat (Ondatra zibethicus) found moribund in Illinois (USA) had minimal meningitis and pleuritis, probably of bacterial origin. There were large, basophilic, intranuclear inclusion bodies within scattered enterocytes. The inclusions were microscopically typical of those produced by adenoviruses, and ultrastructurally were intranuclear paracrystalline arrays of virus particles with characteristics of adenoviruses. The significance of the adenovirus infection in this muskrat is unknown. PMID- 11504245 TI - Reference values and limited serological survey for the Iriomote cat in Japan. AB - Clinical investigations and hematological, serum biochemical, and serological surveys were carried out on 11 male and 6 female Iriomote cats (Felis iriomotensis) in Japan. Examined Iriomote cats were considered clinically healthy by the inspection for the general physical conditions. However, urinalysis suggested the inflammation of the urinary tract in all the cats. Antibody for feline panleukopenia virus was positive in one of the examined Iriomote cats, which suggested a previous infection. PMID- 11504244 TI - Fecal corticosterone reflects serum corticosterone in Florida sandhill cranes. AB - Florida sandhill cranes (Grus canadensis pratensis) were conditioned to confinement 6 hr/day for 7 days. On day 8, each bird's jugular vein was catheterized, blood samples were drawn, and each crane was confined for 6 hr. Using a randomized, restricted cross-over design, cranes were injected intravenously with either 0.9% NaCl solution or ACTH (cosyntropin; Cortrosyn; 0.25 mg). During the 6 hr of confinement, fecal samples (feces and urine) were collected from each of five cranes immediately after defecation. Individual fecal samples were collected approximately at hourly intervals and assayed for corticosterone. We showed previously that serum corticosterone did not vary significantly following saline injection, but peaked significantly 60 min after ACTH injection. Maximal fecal corticosterone concentrations (ng/g) were greater (P < 0.10; median 1087 ng/g) following ACTH stimulation compared to maximal fecal corticosterone concentrations at the end of acclimation (day 7; median 176) and following saline treatment (median 541). In cranes under controlled conditions, fecal corticosterone concentration reflects serum corticosterone levels, fecal corticosterone, Grus canadensis pratensis, sandhill cranes, serum corticosterone levels. PMID- 11504246 TI - Aflatoxin levels in corn available as wild turkey feed in Georgia. AB - Samples of corn available as wildlife feed from retailers throughout Georgia (USA) were collected during April 1997 and analyzed for aflatoxin to determine if levels harmful to wild turkeys (Meleagris gallopavo) were present. Three of 31 (10%) samples collected from a 40-country area were positive. An enzyme-linked immunosorbent assay qualitatively determined that two samples contained from 0 to 20 ppb aflatoxin. A chromatography analysis of a third sample measured 380 ppb total aflatoxin. A small percentage of our sample of wildlife feed collected during one season contained levels of aflatoxin that may cause harm to turkeys, especially poults. However, because aflatoxin levels ranging from 100 to 400 ppb may cause liver dysfunction and immunosuppression in turkey poults and other wildlife, grains known to be contaminated with aflatoxin at levels unacceptable for domestic animal feeds (> or =100 ppb) should not be sold as wildlife feed. Further analyses of grains sold as wildlife feed should be conducted to address this potential problem. PMID- 11504247 TI - Anatomical variations in the coronary arteries. AB - The following are observations on 500 consecutive cases of coronary angiographical studies carried out on patients without any case history of myocardial ishcemia. There were 251 males and 249 females. The patients' average age was 45 years old. PMID- 11504248 TI - High division of the median nerve in the carpal tunnel: anatomical observations. AB - Here are analysed personal observation about high duplication of median nerve at the wrist. Here are reported 18 cases on 3300 operations for carpal tunnel. The results are compared with the literary casuistry. PMID- 11504249 TI - The nerves of the upper and lower extremities which are at a greater risk of compressive pathology. Anatomical topographical correlations. Our experience. AB - The experience of numerous cases of surgical operations carried out over the ten years from 1990-2000, has meant we have been able to reflect on the fact that the median, ulnar, radial (upper extremity), tibial and external popliteal sciatic (lower extremity) nerves are more greatly exposed to compressive pathology. This is due to certain anatomical topographical relationships with canals, semi canals, tunnels in well-defined and unchanging sites. From this point of view, a pathology with an etiopathogenic topographical tendency is emerging. PMID- 11504250 TI - Functional anatomy and imaging of the foot. AB - The foot is constituted from a series of small bones making a segmented structure with multiple joints, likened to a dome, in contact with the ground in three points: posteriorly the calcanear tuberosity; anteriorly and medially the head of 1st metatarsum, and anteriorly and laterally the head of 5th metatarsum. In fact, each foot presents a semi-arch whose base is represented by the lateral border and the summit by the medial border of the foot. The foot has been likened to a half-dome, so that when the medial borders of the two feet are placed together, a complete dome is formed. In the foot are present two longitudinal arches: the medial consists of the calcaneus, the talus, the navicular, the three cuneiform bones and the first three metatarsal bones. It is more arcuated and elastic then the lateral, that consist of the calcaneus, the cuboid and the 4th and 5th metatarsus. This is flattened and in contact with the ground. We can identify two transverse arches between longitudinal arches, extending from the medial to the lateral borders of the foot: the first is a lancet dome, between midfoot and forefoot, at the tarsometatarsal joint level; it consists of the bases of the metatarsal bones and the cuboid and the three cuneiform bones; the second is at flat dome, in correspondence of forefoot, at the metatarsophalangeal joint level; it consists of the bases of the proximal phalanges of the fingers and the head of five metatarsal bones. Longitudinal and transversal arches are supported from: the shape of stones of the structure (foot bones); the long and short plantar ligaments, larger and stronger than the dorsal ligaments, tie together the lower edges of the bones; a beam (the plantar aponeurosis and the plantar muscles and tendons) connecting the ends of the bridge effectively prevents separation of the pillars and consequent sagging of the arch; the maintenance of the arch depends on multiple support (ligaments, tendons of extrinsic muscles of the foot) suspending the arch from a cable above the level of the bridge. In this manner, we obtain a "functional unit" with two important aims: to support the body weight (static foot), and to serve as a lever to propel the bodies forward in walking and running (dinamic foot). Imaging techniques are able to provide further data on functional anatomy of the foot; in particular, these techniques show the bones structures, ligaments, muscles and tendons, taking part to the arch setting. Routine x-ray examinations precise information about the bone-shape and their relationship. The short and long ligaments, the tendons and the suspending the arch from above are instead successfully valuable using ultrasonography, CT and MR. PMID- 11504252 TI - Evidence-based practice: new techniques and technology. PMID- 11504251 TI - Symmetry of healthy adult feet: role of orthostatic footprint at computerized baropodometry and of digital formula. AB - Morphological aspects of orthostatic footprints (anterior heel, isthmus, posterior heel), using computerized baropodometry and of the digital formula were studied in 97 subjects (37 males, 60 females; median age: 20.9+/-1.56) at the Medical School of Palermo, Italy. The aim of this study was to contribute to our knowledge of the bilateral symmetry and asymmetry of the human feet in ethnically similar groups. We evaluated the length of the footprints (FL) and the widths of the anterior heel (AHW), isthmus (IW), and the posterior heel (PHW). Values were compared in the left and right feet of each subjects. The general morphology of the footprints was considered to determine bilateral correspondence or divergence. We also evaluated the digital formula to verify any bilateral correspondence. The linear measurements of the footprints did not show any particular bilateral conformity. The general morphology of the footprint showed bilateral correspondence in 76 subjects (78%): in 21 subjects (22%) it did not show any. Typological results of the bilateral orthostatic footprints showed normalfootprints in 54 subjects (55.5%): the isthmus included 1/3 to 2/3 of the AHW. Hollow footprints were found in 20 subjects (20.5%): the isthmus was less than 1/3 of the AHW. At clinical examination, using the digital formula, we found that Egyptian foot (1 degree > 2 degrees) was the most frequent (68%). The standard foot (1 degree = 2 degrees) and the Greek foot (1 degree < 2 degrees) were both present in 16% of the feet examined. Seventy-nine subjects (82%) presented bilaterally correspondent digital formulas: 1 degree > 2 degrees in 59 subjects (62%), 1 degree = 2 degrees in 9 subjects (9%), and 1 degree < 2 degrees in 11 (11%). While comparison of the bilateral linear measurements studied showed that there was primarily no bilateral correspondence, the morphological aspects of the footprints and of the digital formulas showed that bilateral correspondence was prevalent. PMID- 11504253 TI - SESAP questions. Laceration of the perineum. PMID- 11504254 TI - Interrator reliability in osteoarthritis. PMID- 11504255 TI - Musculoskeletal images. Desmoplastic fibroma of the thigh. PMID- 11504256 TI - Soft-tissue images. Bullet embolism. PMID- 11504257 TI - Soft-tissue case 40. Boerhaave's syndrome: postemetic esophageal rupture. PMID- 11504258 TI - Musculoskeletal case 17. Dislocation of the lunate bone. PMID- 11504259 TI - Efficacy of calcium sulfate plus decompression bone in lumbar and lumbosacral spinal fusion: preliminary results in 40 patients. AB - OBJECTIVE: To compare the efficacy of calcium sulfate pellets plus bone obtained from decompression with fresh autologous iliac crest bone in lumbar and lumbosacral spinal fusion with decompression. DESIGN: A prospective randomized clinical trial. SETTING: Four tertiary care centres in Halifax, NS. PATIENTS: All were consenting, skeletally mature and suffering from degenerative disc disease or spondylolisthesis. The first 40 patients from a single test centre are reported; 32 of these had completed 1-year follow-up. INTERVENTIONS: Posterolateral lumbar or lumbosacral spinal fusion with spinal canal decompression and randomized placement of test material (decompression bone plus an equal volume of calcium sulfate pellets) on one side and control material (autologous posterior iliac crest bone of equal volume to the test material) on the contralateral side, which allowed subjects to act as their own control. OUTCOME MEASURES: Assessment of bone formation by radiographic evaluation at 6 and 12 months after fusion by an independent musculoskeletal radiologist blinded to the placement of test material. RESULTS: At 6 and 12 months after fusion, 78% and 88% of patients, respectively, showed bone formation at the test site that was 75% to 100% of, equal to or more than that at the control site. Increases in bone formation at 6 and 12 months were almost identical at both sites. Smoking status, patient gender or age, instrumentation used and volume of graft used were not predictive of outcome. CONCLUSIONS: Calcium sulfate pellets plus decompression bone provided bone formation equivalent to autologous iliac crest bone in a majority of patients. Calcium sulfate pellets plus decompression bone may provide a viable alternative to autologous iliac crest as a graft material for spinal fusion. PMID- 11504260 TI - Physicians' accuracy and interrator reliability for the diagnosis of unstable meniscal tears in patients having osteoarthritis of the knee. AB - OBJECTIVE: To determine clinicians' accuracy and reliability for the clinical diagnosis of unstable meniscus tears in patients with symptomatic osteoarthritis of the knee. DESIGN: A prospective cohort study. SETTING: A single tertiary care centre. PATIENTS: One hundred and fifty-two patients with symptomatic osteoarthritis of the knee refractory to conservative medical treatment were selected for prospective evaluation of arthroscopic debridement. INTERVENTION: Arthroscopic debridement of the knee, including meniscal tear and chondral flap resection, without abrasion arthroplasty. OUTCOME MEASURES: A standardized assessment protocol was administered to each patient by 2 independent observers. Arthroscopic determination of unstable meniscal tears was recorded by 1 observer who reviewed a video recording and was blinded to preoperative data. Those variables that had the highest interobserver agreement and the strongest association with meniscal tear by univariate methods were entered into logistic regression to model the best prediction of resectable tears. RESULTS: There were 92 meniscal tears (77 medial, 15 lateral). Interobserver agreement between clinical fellows and treating surgeons was poor to fair (kappa < 0.4) for all clinical variables except radiographic measures, which were good. Fellows and surgeons predicted unstable meniscal tear preoperatively with equivalent accuracy of 60%. Logistic regression modelling revealed that a history of swelling and a ballottable effusion were negative predictors. A positive McMurray test was the only positive predictor of unstable meniscal tear. "Mechanical" symptoms were not reliable predictors in this prospective study. The model was 69% accurate for all patients and 76% for those with advanced medial compartment osteoarthritis defined by a joint space height of 2 mm or less. CONCLUSIONS: This study underscored the difficulty in using clinical variables to predict unstable medial meniscal tears in patients with pre-existing osteoarthritis of the knee. The lack of interobserver agreement must be overcome to ensure that the findings can be generalized to other physician observers. PMID- 11504261 TI - Toe-to-finger transfer for post-traumatic reconstruction of the fingerless hand. AB - OBJECTIVE: To assess the utility of toe-to-finger transfers (TFTs) for post traumatic reconstruction of the fingerless hand. DESIGN: A case series. SETTING: A regional trauma centre. PATIENTS: Eight men, mean age was 36 years (range from 25-59 yr), who had lost all the fingers from a hand due to a crush-degloving injury (6 patients), frostbite (1 patient) or a burn injury (1 patient). INTERVENTION: TFT. Twelve TFTs were cone and the mean time from injury to reconstruction was 17.2 months. MAIN OUTCOME MEASURES: Objective (range of motion, moving 2-point discrimination, grip strength, key pinch, Jebsen-Taylor hand assessment, return to work) and subjective (activities of daily living and a questionnaire) measures. RESULTS: Eleven of the 12 transfers survived. Six of the 7 in whom the transfer was successful were available for follow-up (mean 45 mo). Range of motion was 10 degrees at the distal interphalangeal joint, 18 degrees at the proximal interphalangeal joint and 59 degrees at the metacarpophalangeal joint. Sensation was protective in all. Grip strength and key pinch were 26.1% and 70.2% of the contralateral hand respectively. Jebsen-Taylor assessment indicated that basic activities were possible but slowed. All 6 patients returned to work and could perform 92.6% of the activities of daily living unassisted. Hand and foot symptoms were mild. Two-thirds were appearance conscious, 5 of the 6 went on to altered vocations and all reported overall satisfaction as high. CONCLUSION: This study supports TFT for reconstruction of the fingerless hand in that, although transferred toe function may be poorer than a normal finger, the hand is restored to a useful, sensate and versatile functional unit, such that global hand and patient function, as well as patient satisfaction, are very good. PMID- 11504262 TI - Empyema thoracis: lack of awareness results in a prolonged clinical course. AB - OBJECTIVE: To assess the hypothesis that empyema thoracis (ET) is a problem often not optimally treated. Long delays in diagnosis are common, long hospital stays are typical and recovery with surgery is relatively rapid. DESIGN: A chart review. SETTING: The Regina Health District associated hospitals, a tertiary referral centre. PATIENTS: The charts of 34 consecutive patients having primary respiratory tract disease and seen during the 6-year period Apr. 1, 1991, to Mar. 31, 1997, were identified. OUTCOME MEASURES: Patient presentation, time until diagnosis of ET, number of radiologic investigations, microbiologic features, treatment methods, postoperative course and mortality. RESULTS: The mean delay in diagnosis, defined as the time of admission to the time of correct diagnosis, was 44.2 days (range from 0 to 573 days) and the mean delay until thoracic surgery referral was 47.4 days (range from 0 to 578 days). On average each patient underwent CT 10.1 times, had 2.6 percutaneous drainage procedures and 2.0 chest tube insertions. The mean time from the first percutaneous chest drainage to the date of diagnosis was 29.8 days (range from 0 to 564 days). Of the 26 patients who underwent CT, the mean time from the first CT of the chest to the date of diagnosis was 9.5 days (range from 0 to 75 days). Cultures of pleural fluid grew no organisms in 17 patients; in the remaining 17 patients cultures grew 23 different microorganisms. Of 26 patients who were referred for surgical opinion, 18 underwent decortication; 8 were not considered to be surgical candidates. Pathological examination showed 17 cases of inflammatory empyema and 1 case of mesothelioma (unrecognized clinically). The mean length of hospital stay postoperatively was 15.2 days. CONCLUSIONS: Early suspicion of ET facilitates its treatment, resulting in fewer investigations and shorter hospital stays. When percutaneous drainage does not eliminate pleural effusions, empyema must be considered. Recovery from surgical decortication is rapid in comparison with the typical protracted preoperative hospital course. PMID- 11504263 TI - Extra-articular deformity in distal radial fractures treated by external fixation. AB - OBJECTIVE: To determine the radiographic outcome in fractures of the distal radius treated with closed reduction and external fixation. DESIGN: A retrospective study. SETTING: The orthopedic department of National Taiwan University Hospital. PATIENTS: Eighty-five consecutive patients (36 female, 49 male), average age 48 years, with fractures of the distal radius seen between March 1995 and June 1998. INTERVENTIONS: Closed reduction and external fixation of fractures, followed up by good-quality posteroanterior and lateral radiographs to evaluate healing. MAIN OUTCOME MEASURES: Radial height, radial inclination and volar tilt were measured on radiographs obtained initially, immediately postoperatively and at the time of removal of external fixation. Data were analyzed by the t-test. RESULTS: Immediately after reduction and fixation, there was a significant improvement in the measurements of radial height and radial inclination. This improvement was gradually lost and height and inclination were significantly decreased at the time the external fixation device was removed. External fixation did not improve the volar tilt. CONCLUSION: External fixation is a popular method to improve the reduction of osseous deformity but cannot effectively protect comminuted distal radial fractures from loss of reduction, which may be associated with shortening and redisplacement. PMID- 11504264 TI - Re-expansion pulmonary edema after repair of a missed diaphragmatic hernia. PMID- 11504265 TI - Vernix caseosa peritonitis presenting post partum as acute cholecystitis. PMID- 11504266 TI - Post-traumatic nonunion of the ulna in a child. PMID- 11504267 TI - Anastomotic stenosis after minimally invasive direct coronary artery bypass. PMID- 11504268 TI - Population and individual bioequivalence: lessons from real data and simulation studies. AB - The Food and Drug Administration (FDA) has proposed replacing the 1992 average bioequivalence (ABE) with population and individual bioequivalence (PBE & IBE), as outlined in the preliminary draft guidance of December 1997, which was subsequently replaced by the draft guidances of August 1999 and resolved in the final guidance of October 2000. This has led to considerable public debate among regulatory, academic, and industry experts at numerous conferences (e.g., FDA/AAPS March 1998, FDA/AAPS August-September 1999, FDA Pharmaceutical Sciences Advisory Committee September 1999) and in the literature. The final guidance calls for ABE to remain as the primary criterion by which new formulations may be judged ready for access to the marketplace. In addition, the FDA recommends the use of replicate study designs for the specific drug classes of controlled release formulations and highly variable drugs. The final guidance also alludes to the possibility of a sponsor requesting alternative criteria such as PBE and IBE following consultation with the FDA. This procedure amounts to a data collection period during which data suitable to evaluate the operating characteristics of PBE and IBE would be generated, analyzed, and discussed among interested parties. A comprehensive review of currently available databases is useful in determining the ultimate value of this data collection period. This report provides an update to the previous publication by the authors. In all, 28 data sets from 20 replicate cross-over bioequivalence studies have been analyzed (n = 12-96) using the statistical methodology in the most recent FDA draft guidance. The results are presented below. ABE Pass: ABE Fail: Total: AUC/Cmax AUC/Cmax AUC/Cmax AUC/Cmax Pass PBE & IBE 20/14 1/3 21/17 Pass IBE only 1/0 0/0 1/0 Fail PBE and IBE 0/2 0/1 0/3 Fail IBE only 2/3 4/5 6/8 Total 23/19 5/9 28/28 Review of the database reveals many interesting features, most notably the lack of consistent results within a given data set across all three criteria. The sensitivity of subject-by-formulation interaction to sample size and inherent variability of the compounds is further explored through simulation studies. It is concluded that additional simulation assessments must be considered when evaluating the value of a data collection period for PBE and IBE assessment. It will be shown that definitive conclusions regarding some of the operating characteristics of PBE and IBE can be achieved through a combination of data driven hypotheses followed by simulation studies to further evaluate the hypotheses. Some recommendations for further data collection will be made. PMID- 11504269 TI - Contributions of five human cytochrome P450 isoforms to the N-demethylation of clozapine in vitro at low and high concentrations. AB - The authors assessed the in vitro contribution of cytochrome P450 (CYP) isoforms 1A2, 3A4, 2C9, 2C19, and 2D6 to the N-demethylation of clozapine mediated by human liver microsomal preparations (HLM). In contrast to previous studies, the authors focused on a relatively low hepatic concentration level, 5 microM, to assess the conditions at a therapeutically relevant hepatic concentration level of clozapine. The optimal concentrations of specific inhibitors were initially established using cDNA-expressed CYP isoforms. The mean contributions of CYPs 1A2, 2C19, 3A4, 2C9, and 2D6 amounted to 30%, 24%, 22%, 12%, and 6%, respectively, with regard to the total HLM-mediated N-demethylation. Thus, the present in vitro study on clozapine N-demethylation suggests that CYP1A2 is the most important form at low concentrations, which is in agreement with clinical findings. CYP2C19 is also of considerable importance, while the roles of CYP2C9 and 2D6 are more modest. CYP3A4 attained a dominating role with an average contribution of 37% at a high clozapine concentration (50 microM). The rate of other metabolic routes mediated by CYP2D6 only corresponded to about one fifth of the CYP2D6 catalyzed N-demethylation rate. PMID- 11504270 TI - Oral bioavailability and disposition of [14C]omapatrilat in healthy subjects. AB - The objective of this study was to determine the absolute oral bioavailability and disposition of omapatrilat. This single-dose, randomized, crossover study of 20 mg intravenous and 50 mg oral [14C]omapatrilat was conducted in 12 healthy male subjects to determine the disposition and oral bioavailability of omapatrilat, an orally active vasopeptidase inhibitor. Blood samples were collected up to 120 hours, and the excreta were collected over 168 hours postdose. Plasma concentrations of omapatrilat were determined by a validated LC/MS/MS procedure. Radioactivity in blood, plasma, urine, and feces was determined by liquid scintillation counting. Urinary excretion of radioactivity averaged 80% and 64% of intravenous and oral doses, respectively; < 1% of oral dose was excreted unchanged in urine. The absolute oral bioavailability of omapatrilat averaged 31%. Total body clearance of omapatrilat (80 L/h) exceeded liver plasma flow. Apparent steady-state volume of distribution of omapatrilat (21 L/kg) was extremely high compared with total body water. Omapatrilat undergoes substantial presystemic first-pass metabolism after oral administration. Omapatrilat is eliminated primarily by metabolism, and its metabolites are eliminated primarily in urine. Extrahepatic organs may be involved in the elimination of omapatrilat. Plasma concentrations of omapatrilat exhibit a prolonged terminal elimination phase, which represents elimination from a deep compartment. PMID- 11504271 TI - The safety of higher than standard dose of doxylamine-pyridoxine (Diclectin) for nausea and vomiting of pregnancy. AB - A delayed-release combination of doxylamine-pyridoxine (D-P) (Diclectin) is the only approved antiemetic medication for use in pregnancy in Canada. The standard recommended dose is up to 4 tablets a day, regardless of body weight or severity of symptoms. The objective of this study was to determine the incidence of adverse maternal and fetal effects and pregnancy outcome in 225 women taking Diclectin at the recommended (n = 123) or higher than recommended (n = 102) doses. In this observational, prospective study, one-third (33.6%) of women reported having adverse effects (sleepiness, tiredness, and/or drowsiness) temporally related to the medication. There was no association between the dose per kg and rates of reported maternal adverse effects with doses ranging from 0.1 mg/kg to 2.0 mg/kg (1-12 tablets). Nausea and vomiting of pregnancy (NVP) was reported as severe by the majority (75.8%) of women. Mean birth weight (BW) was 3,400 g and gestational age (GA) 39 weeks. Multivariate analysis revealed that only prepregnancy weight and GA predicted lower BW, not the dose of D-P or the severity of NVP. There were two pregnancies with major malformation, a finding that is consistent with the rates of birth defects in the general population. It was concluded that the higher than standard dose of Diclectin, when calculated per kg of body weight, does not affect either the incidence of maternal adverse effects or pregnancy outcome. If needed, Diclectin can be given at doses higher than 4 tablets/day to normalize for body weight or optimize efficacy. PMID- 11504272 TI - Measurement of acetaminophen-protein adducts in children and adolescents with acetaminophen overdoses. . AB - Acetaminophen-protein adducts are biomarkers of acetaminophen toxicity present in the centrilobular region of the liver of laboratory animals following the administration of toxic doses of acetaminophen. These biomarkers are highly specific for acetaminophen-induced hepatic injury and correlate with hepatic transaminase elevation. The objective of this prospective, multicenter study was to evaluate the clinical application of the measurement of acetaminophen-protein adducts in pediatric acetaminophen overdose patients. Serum samples were obtained from 51 children and adolescents with acetaminophen overdose at the time of routine blood sampling for clinical monitoring. Six subjects developed "severe" hepatotoxicity (transaminase elevation > 1,000 IU/L), and 6 subjects had transaminase elevation of 100 to 1,000 IU/L. Acetaminophen-protein adducts were detected in the serum of only 1 study subject, a patient with marked transaminase elevation (> 6,000 IU/L) and high risk for the development of hepatotoxicity according to the Rumack nomogram. While this study provides further support for the occurrence of covalent binding of acetaminophen to hepatic protein in humans following acetaminophen overdose, the detection of acetaminophen-protein adducts in serum with the current methodology requires significant biochemical evidence of hepatocellular injury. PMID- 11504273 TI - The risk of liver damage associated with minocycline: a comparative study. AB - Using the General Practice Research Database, the authors performed (1) a cohort analysis comparing the incidence of liver dysfunction in new users of minocycline with new users of oxytetracycline/tetracycline and (2) a case control study assessing antibiotic exposure in new cases of liver dysfunction and controls without liver dysfunction. In new users, the incidence of liver dysfunction in those exposed to minocycline was 1.04 cases/10,000 exposed person months (EPM) and 0.69 cases/10,000 EPM in those exposed to oxytetracycline/tetracycline (relative risk 1.51 [CI95: 0.63, 3.65]). The risk in both groups was greatest in the first month of use. The adjusted odds ratio (ORadj) of liver dysfunction associated with exposure to minocycline compared with nonuse was 2.10 (CI95: 1.30, 3.40); for oxytetracycline/tetracycline, the ORadj was 1.46 (CI95: 0.81, 2.64); and for exposure to erythromycin, the ORadj was 1.64 (CI95: 0.71, 3.80). The authors thus support a weak association between the use of oral antibiotics and liver dysfunction in patients with acne. The risk associated with exposure to minocycline appears to be very small. The cohort analysis demonstrated that any risk associated with minocycline was not significantly greater than that associated with oxytetracycline/tetracycline exposure. PMID- 11504274 TI - Comparison of azathioprine and mycophenolate mofetil for the prevention of acute rejection in recipients of pancreas transplantation. AB - The study was performed to compare the efficacy and side effects of azathioprine (AZA) and mycophenolate mofetil (MMF) in conjunction with cyclosporine or tacrolimus and steroids for the prevention of acute pancreas rejection during the first 6 months of pancreas transplantation. In this case-controlled study, MMF is compared with historical controls of AZA in the prevention of acute pancreas rejection. The primary measures of treatment efficacy were patient and pancreas survival rate at 6 months after transplantation. Secondary efficacy measures were the occurrence of biopsy-proven pancreas rejections and the use of antilymphocyte preparations for rejection treatment. A total of 111 pancreas transplant patients (57 in the AZA group and 54 in the MMF group) were evaluated. The 6-month patient survival rate was 96% in the AZA group versus 97% in the MMF group (p = 0.57). The 6-month pancreas graft survival rate was 88% in the AZA group versus 91% in the MMF group (p = 0.29). However, biopsy-proven rejection episodes during the first 6 months of transplantation were significantly lower with MMF (46%) than with AZA (69%) (p = 0.01). In addition, patients in the AZA group received a greater number of full courses of antilymphocyte therapy as a rejection treatment (p = 0.004). Overall, the frequency of adverse events was similar, although the MMF group experienced higher incidences of gastrointestinal adverse events. In conclusion, compared with AZA, MMF significantly reduces the rate of biopsy proven pancreas rejection during the first 6 months of transplantation and is well tolerated, except for gastrointestinal adverse events. PMID- 11504275 TI - Intravenous safety and pharmacokinetics of a novel dimerizer drug, AP1903, in healthy volunteers. AB - AP1903 is a novel gene-targeted drug that is being developed for use in drug regulated cell therapies. An intravenous, single-blind, placebo- and saline controlled, ascending-dose study was performed to evaluate the safety, tolerability, and pharmacokinetics of AP1903. Twenty-eight normal healthy male volunteers were randomized into five dosage groups of AP1903 (0.01, 0.05, 0.1, 0.5, and 1 mg/kg). Within each group, 4 volunteers received a single dose of AP1903, 1 volunteer received an equal volume of placebo, and 1 received an equal volume of normal saline. The only exception was in the 0.5 mg/kg group, in which 4 volunteers were dosed: 3 received AP1903 and 1 received normal saline. All dosages were administered as intravenous infusions over 2 hours. Clinical safety parameters were monitored, and serial blood and urine samples were collected for analysis of AP1903. No drug-related adverse events were observed at any of the dose levels with the possible exception of facial flushing in 1 volunteer at the 1.0 mg/kg dose level. AP1903 plasma levels were directly proportional to the administered dose, with mean Cmax values ranging from approximately 10 to 1,275 ng/mL over the 0.01 to 1.0 mg/kg dose range. Following the infusion period, blood concentrations revealed a rapid distribution phase, with plasma levels being reduced to approximately 18%, 7%, and 1% of the maximal concentration at 0.5, 2, and 10 hours postdose, respectively. AP1903 was shown to be safe and well tolerated at all dose levels and demonstrated a favorable pharmacokinetic profile at doses well above the anticipated therapeutic dose. PMID- 11504276 TI - Pharmacokinetics and safety of FFR-rFVIIa after single doses in healthy subjects. AB - FFR-rFVIIa is an antithrombotic agent, which has also proven to have antirestenotic properties in animal models. FFR-rFVIIa is a modified recombinant FVIIa in which the catalytic site is irreversibly inactivated by a synthetic tripeptide covalently bound with the FVIIa molecule. The modified rFVIIa retains its tissue factor (TF) binding capacity but is otherwise enzymatically inactive. A double-blind, placebo-controlled, randomized dose escalation trial was conducted to investigate eight single i.v. doses of FFR-rFVIIa (0.01, 0.02, 0.05, 0.08, 0.12, 0.18, 0.27, or 0.40 mg/kg body weight) in healthy male volunteers (n = 62). Safety, pharmacokinetics, and pharmacodynamics of FFR-rFVIIa were assessed. Mean (SD)AUC0-infinity ranged from 0.35 (0.11) to 28.8 (3.5)microg.h/ml, and mean Cmax ranged from 0.078 (0.019) to 4.8 (0.7) microg/ml. The mean elimination half-life ranged from 3.8 to 5.8 hours. Mean AUC0-infinity increased with increasing dose levels. Cmax appeared to be proportional to the dose level, with the exception of the lowest dose level. A dose-dependent prolongation of the prothrombin time was found, demonstrating that FFR-rFVIIa inhibited coagulation via the TF-dependent pathway FFR-rFVIIa was generally well tolerated at all dose levels studied. PMID- 11504277 TI - A comparative multidose pharmacokinetic study of buspirone extended-release tablets with a reference immediate-release product. AB - Buspirone is disclosed in U.S. Patent No. 3,717,634 as a pharmaceutically active compound that has been found to be effective for the treatment of anxiety disorders and depression. In this randomized, two-treatment, two-period, multidose crossover study, the pharmacokinetics of a once-daily extended-release (ER)formulation of buspirone was compared with that of an immediate-release (IR) formulation of commercially available buspirone. A total of 30 mg of the ER formulation was administered to 36 healthy volunteers once daily for 7 days, and 15 mg of the IR formulation was administered twice daily for 7 days. Pharmacokinetic profiles of buspirone and its metabolite, 1-pyrimidinylpiperazine (1-PP), were obtained at steady state. The bioavailability of buspirone from the ER formulation was more than three times higher than that from the IR formulation at steady state, and that of 1-PP was about 25% less. The mean steady-state Cmax of buspirone from the ER formulation was 46% higher than that from the IR formulation (p < 0.05), and that for 1-PP was lower by 29% (p < 0.05). The mean apparent half-life of buspirone from the ER formulation (9.04 hours) was considerably longer than that observed for the IR formulation (3.06 hours). The median 1-PP/buspirone AUC ratio was much higher for the IR formulation at steady state (24.4) than for the ER formulation (6.44). There were no significant differences in average pharmacokinetic metrics observed in men and women. Based on these observations of the potential benefits of once-daily dosing with the ER product in terms of prolonged buspirone plasma concentrations, a significant increase in the ratio of buspirone to 1-PP concentration with a lower intersubject variation could be achieved that should provide an improvement in the desired therapeutic effects of buspirone. PMID- 11504278 TI - Steady-state pharmacokinetics and tolerability of modafinil given alone or in combination with methylphenidate in healthy volunteers. AB - The potential for a pharmacokinetic (PK) drug-drug interaction between modafinil and methylphenidate, each at steady state, was investigated in an open-label, randomized, single-period study in 32 healthy male and female volunteers. All subjects received modafinil once daily orally for 28 days (200 mg on Days 1-7 and 400 mg on Days 8-28). On Days 22 to 28, half of the subjects also received 20 mg of methylphenidate orally 8 hours after their modafinil dose. PK profiles of modafinil were obtained on Days 21 and 28 and compared between the two groups. There were no statistically significant differences between the treatment groups in the mean changes in PK parameters for modafinil. Parameters for its metabolites were also similar between the groups, and all treatments were well tolerated. The results indicate that administration of low-dose methylphenidate 8 hours after treatment with modafinil does not appear to alter the steady-state pharmacokinetics of modafinil in healthy volunteers. PMID- 11504279 TI - Current treatment approaches for chronic myelogenous leukemia. AB - Chronic myelogenous leukemia (CML) is a clonal hematopoietic stem cell disorder that progresses through distinct phases characterized by progressive loss of the differentiation of the malignant clone. Over the past 4 decades, it has been established that the Bcr-Abl protein, created as a consequence of a (9:22) chromosomal translocation, is the cause of the disease. Bcr-Abl functions as a constitutively activated tyrosine kinase, and this kinase activity is absolutely required for the transforming function of the Bcr-Abl protein. Thus, a specific inhibitor of the Bcr-Abl tyrosine kinase would be predicted to be an effective and selective therapeutic agent for CML. STI571, an Abl-specific tyrosine kinase inhibitor, has shown remarkable activity in all phases of CML. The clinical features, molecular pathogenesis, and current treatment options of CML are reviewed along with the development of STI571, the phase I clinical results, and the application of this paradigm to other malignancies. PMID- 11504280 TI - Novel biologically based therapies for myeloma. AB - Recent advances in our understanding of the molecular regulation of myeloma cells suggest novel strategies for treating multiple myeloma. Some myeloma cells express a 69 kD variant of Ku86, a heterodimer subunit that is essential for double-stranded DNA break repair. Presence of the variant impairs DNA repair; therefore normal Ku86 in myeloma cells confers resistance to therapy and may represent a therapeutic target. The upregulation of NF-kappaB-dependent interleukin-6 (IL-6) transcription and secretion that occurs following adhesion of myeloma cells to bone marrow stromal cells (BMSCs) may serve as a potential therapeutic target, as IL-6 is a growth and survival factor for myeloma cells. Accordingly, proteasome inhibitors inhibit activation of NF-kappaB and induce apoptosis of myeloma cells; they also inhibit the NF-kappaB-dependent up regulation of IL-6 in BMSCs and related paracrine growth of adherent tumor cells. Therapeutic strategies may also target the mitogen-activated protein kinase (MAPK) pathway that is thought to mediate the IL-6-induced proliferation of myeloma cells. Vascular endothelial growth factor (VEGF) is also upregulated by adhesion of myeloma cells to BMSCs and may serve as a growth and/or survival factor for myeloma cells; preliminary studies suggest that VEGF receptor inhibitors may block proliferation of tumor cells. Thalidomide was recently used successfully to treat myeloma in patients whose disease was refractory to conventional treatment. An enhanced understanding of the mechanisms of action of thalidomide may result in the development of analogues with enhanced potency and fewer side effects. The potential mechanisms of action of thalidomide are reviewed, including antiangiogenic effects; direct effects of thalidomide on the growth and survival of myeloma cells and BMSCs; modulation of adhesive interactions; and regulation of secretion and bioactivity of cytokines. Immune based strategies for treating multiple myeloma are also reviewed. Therapeutic obstacles include excessive toxicity after allografting, contaminating tumor cells in autografts, and the persistence of minimal residual disease (MRD) after high-dose therapy followed by allogenic or autologous stem cell transplantation. Allografting can be performed safely in myeloma, donor lymphocyte infusions (DLI) may effectively treat relapsed myeloma post allografting; and use of CD4+ T cell enriched DLI may reduce the risk of graft-versus-host disease. Treatment with autografting is frequently compromised by MRD in the autograft and in the patient post myeloablative therapy. Adenoviral purging prior to autotransplantation and in vivo and ex vivo stimulation of autoimmune cells are discussed as potential approaches to address these problems. PMID- 11504281 TI - National Oncology Forum: perspectives for the year 2000. AB - Despite advances in treatment, long-term outcome of patients with diffuse large B cell lymphoma (DLBCL) is no better today than reported in 1975. A recent study applying DNA microarray technology revealed that patients whose cancer related to patterns of genes expressed in germinal center lymphocytes responded more favorably to chemotherapy than patients whose cancer related to patterns of genes expressed in activated lymphocytes. cDNA and oligonucleotide microarrays are described, and their applications in cancer research are reviewed. In addition to DLBCL, microarray technology has been used to study several types of cancer. The applications of microarray technology are numerous and include profiling gene expression patterns in order to facilitate diagnosis and predict response to therapy; correlating patterns of gene expression with prognosis; and identifying genes and gene products that are associated with tumorigenic phenotype or with drug resistance, among other applications. Microarraytechnology has also been used in cell lines to correlate gene expression and chemotherapy response. Furthermore, microarray technology may provide a useful tool to examine the development of drug resistance in cancer and has recently been used to study changes in gene expression caused by activated c-Myc in primary human fibroblasts. Tissue microarrays are described. In addition to the amplification of limited tissue re sources, tissue microarrays have the advantages of limiting the variability associated with tissue processing and limiting the necessary amount of reagent. Tissue microarrays have been used to determine the frequencies of amplication of 3 major breast cancer genes and identify overexpression of ERBB2 mRNA; assess and compare gene amplification in benign prostatic hyperplasia, primary prostate carcinoma, recurrent prostate tumors, and metastatic tumors; compare aggressiveness of prostate carcinoma in 2 patient populations; and study gene amplification across various tumor types. Furthermore, DNA microarray and tissue microarray techniques can be combined to provide convergent evidence of findings and to examine different aspects of gene expression. DNA array technology may also be used to identify critical molecular targets or to identify the critical rate-limiting step in a cascade of genes under the influence of a mutated gene. The historical progression of goals of the National Cancer Institute is reviewed, as well as the economic impact of reduction in cancer-associated mortality. Future efforts should continue the investment in basic research and more effectively integrate it with clinical trials and with approaches to prevention and treatment. PMID- 11504282 TI - Adjuvant therapy for melanoma. AB - Patients with deep primaries (> or = 4 mm) or regional lymph node involvement often require adjuvant therapy in addition to surgery to successfully treat melanoma. Various adjuvant strategies are reviewed. Randomized trials of IFN alpha adjuvant therapy have demonstrated statistically significant improvements in disease-free and overall survival rates, leading to approval by the United States Food and Drug Administration of the use of 1 year of intensive IFN-alpha2b following surgical resection of high-risk disease. A study comparing high-dose IFN with the ganglioside vaccine GMK was terminated early when the Data Safety Monitoring Committee concluded that the high-dose IFN treatment arm was associated with highly significantly improved relapse-free and overall survival. Studies of IFN-alpha in stage I and II melanoma are reviewed. Dose and schedule issues in the use of IFN-alpha are addressed. In addition to adjuvant therapy with IFN-alpha, various other treatment strategies appear promising. Adjuvant vaccine therapy may be useful for treatment of cutaneous melanoma. Polyvalent melanoma vaccines are discussed as a potential adjuvant therapy. Finally, nonrandomized preliminary studies suggest that postoperative radiation to the neck or axilla after radical lymph node dissection may decrease regional recurrence rates in node-positive patients, supporting the selective use of radiation therapy for melanoma. PMID- 11504283 TI - Small cell lung cancer: strategies to optimize chemotherapy response. AB - During the past two decades, a major focus of clinical research in small cell lung cancer (SCLC) has been the manipulation of the dose and schedule of the available active cytotoxic agents. Approaches tested include alternating cyclic combination chemotherapy, increasing the dose intensity of chemotherapy with or without the support of either cytokines or stem cells, and increasing the dose density by delivering treatment at shorter intervals. Overall, the results of clinical trials testing these approaches have been disappointing. One of the difficulties in intensifying treatment for SCLC is that the patients tend to be elderly and have smoking-related pulmonary and cardiovascular comorbidities. In fact, as treatment regimens have become more intensive, several multicenter groups have identified a dramatic increase in early-treatment-related mortality rates. Yet, toxicity associated with dose-intensification may obscure a potential therapeutic advantage in unselected patients. Therefore, some of these groups have performed retrospective analyses to identify factors that predict excessive treatment-related toxicity that can be used for patient stratification in clinical trials. This article reviews the data regarding the role of dose intensified therapy in the treatment of SCLC. We propose that delivery of the currently available chemotherapy drugs at greater dose intensity, if excessive toxicity is avoided, may offer a meaningful improvement in survival, and that clinical trials that appropriately test this hypothesis are warranted. PMID- 11504284 TI - Multimodality management of localized pancreatic cancer. AB - Despite improvements in surgical management, only a minority of patients enjoy long-term survival following pancreaticoduodenectomy for adenocarcinoma of the pancreas. Adjuvant therapy consisting of 5-fluorouracil and irradiation has improved median and 5-year survival rates; however, at least one third of eligible patients do not receive postoperative adjuvant therapy because of delayed recovery following pancreaticoduodenectomy. This has led to the development of treatment schedules incorporating the delivery of systemic therapy and/or chemoradiation prior to surgery. This article briefly outlines the history of adjuvant therapy for adenocarcinoma of the pancreas and reviews the available literature on neoadjuvant therapy for localized pancreatic cancer including investigational therapies under clinical trial evaluation. PMID- 11504285 TI - Cervical cancer: combined modality therapy. AB - Prospective, randomized studies conducted over the past 10 years have changed the management of patients with advanced cervical cancer. The reviewed studies evaluated the use of surgery, irradiation, and chemotherapy in patients with various stages of cervical carcinoma in the absence and presence of high-risk factors for recurrence. A study by the Radiation Therapy Oncology Group (RTOG) compared pelvic with pelvic plus prophylactic para-aortic irradiation in patients with stages IB (> 4 cm), IIA, and IIB cervical cancer. The 10-year survival advantage was 11% for patients treated with prophylactic para-aortic irradiation. A follow-up study compared pelvic plus prophylactic para-aortic irradiation and brachytherapy with pelvic irradiation, brachytherapy, and chemotherapy with cisplatin and 5-FU in patients with IB-to IVA-stage cervical cancer. Overall and disease-free survivals were significantly improved in patients receiving chemotherapy. In patients with a prevalence of stage IIB and III, the Gynecologic Oncology Group (GOG) demonstrated that treatment with hydroxyurea alone was inferior to cisplatin or cisplatin, 5-FU, and hydroxy-urea in patients treated concurrently with pelvic irradiation and brachytherapy, and the GOG adopted irradiation and weekly cisplatin as standard therapy. Further GOG studies suggest that irradiation and weekly cisplatin chemotherapy without hysterectomy is the optimal treatment for patients with stage IB cervical cancer. High-risk factors for recurrence include tumor size, depth of tumor invasion, lymphovascular space involvement, and lymph node involvement. Prospective, randomized studies conducted by the GOG evaluated the effectiveness of various treatments in patients with high-risk factors. In one study that did not use chemotherapy, the recurrence-free interval was about 10% better for stage IB patients receiving postoperative irradiation after radical hysterectomy and pelvic lymphadenectomy compared with those who received no further therapy. Patients with Stages IB and IIA disease who, following radical hysterectomy and lymph node dissection, are identified as having positive pelvic lymph nodes and positive parametrial involvement, are at higher risk for recurrence and death than the high-risk group described above. An intergroup study conducted by the GOG, RTOG, and Southwest Oncology Group compared postoperative pelvic irradiation alone with postoperative pelvic irradiation plus concurrent chemotherapy in this group of patients. Overall and progression-free survivals were superior for patients receiving chemotherapy, and their greatest survival occurred in patients who received 3 or 4 chemotherapy cycles compared with 1 or 2 cycles or no chemotherapy. These findings are summarized with respect to their implications fortreatment of patients with advanced cervical cancer. PMID- 11504286 TI - Metabolic and immunologic consequences of laparoscopy with helium or carbon dioxide insufflation: a randomized clinical study. AB - BACKGROUND: Previous studies using animal models have demonstrated that carbon dioxide (CO2) pneumoperitoneum during laparoscopy is associated with adverse physiological, metabolic, immunological and oncological effects, and many of these problems can be avoided by the use of helium insufflation. The present study was performed in patients to compare the effect of helium and CO2 insufflation on intraperitoneal markers of immunological and metabolic function. METHODS: Eighteen patients undergoing elective upper gastrointestinal laparoscopic surgery were randomized to have insufflation achieved by using either helium (n = 8) or CO2 (n = 10) gas. Intraperitoneal pH was monitored continuously during surgery, and peritoneal macrophage function was determined by harvesting peritoneal macrophages at 5 min and 30 min after commencing laparoscopy, and then assessing their ability to produce tumour necrosis factor alpha (TNF-alpha), and their phagocytic function. RESULTS: Carbon dioxide laparoscopy was associated with a lower intraperitoneal pH at the commencement of laparoscopy, although this difference disappeared as surgery progressed. The production of TNF-alpha was better preserved by CO2 laparoscopy, but the insufflation gas used did not affect macrophage phagocytosis. Patients undergoing helium laparoscopy required less postoperative analgesia. CONCLUSION: The choice of insufflation gas can affect intraperitoneal macrophage function in the clinical setting, and possibly acid-base balance. The present study suggested no immunological advantages for the clinical use of helium as an insufflation gas. The outcomes of the present study, however, are different to those obtained from previous laboratory studies and further research is needed to confirm this outcome. PMID- 11504287 TI - Cusum analysis of trends in operative selection and conversion rates for laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) requires a high degree of technical ability, spatial resolution and dexterity. Assessing trainees and competent operators is an important aspect of quality assurance in patient care. Most institutions quote mean conversion rate as a method of comparing operators' performance. The purpose of the present study was to use the technique of cumulative sum (cusum) analysis to determine whether a learning curve phenomenon exists in operators performing LC. METHODS: Data were obtained retrospectively by reviewing the operative records of all patients undergoing elective and acute cholecystectomy for a 30-month period coinciding with the commencement of LC at North Shore Hospital. Patients' age and gender, date and type of operative procedure, duration of operation, and name of operator were recorded. Mean and cusum-transformed data were derived for all operations as well as for four individual operators' performances. RESULTS: Over the study period a total of 614 cholecystectomies was performed, with 85% attempted laparoscopically. A total of 9.8% required conversion to the open technique. Time trend analysis with the cusum technique for all surgeons revealed an inverse relationship between selection rate and conversion rate. Analysis of four individual surgeons revealed three different time trend profiles. CONCLUSION: There was a direct inverse relationship between conversion rate and selection rate, in that careful selection is associated with a low conversion rate. Comparison of individual surgeons' performance showed wide variation, with only one surgeon exhibiting the phenomenon of a learning curve. Contrary to other reports, we found that performance on LC was not always related to operative experience. This highlights the need for a more objective method to analyse operator competence than operator experience alone. PMID- 11504288 TI - Diagnostic accuracy of vacuum-assisted biopsy device for image-detected breast lesions. AB - BACKGROUND: Non-palpable breast lesions present diagnostic difficulties. Ultrasound-guided fine-needle aspiration cytology (FNAC) is a common method used to obtain a diagnosis, but FNAC is frequently inconclusive or insufficiently accurate. Recently a vacuum-assisted biopsy device (Mammotome, Ethicon, Endo surgery, USA) has been introduced. The diagnostic accuracy of this biopsy device was assessed for lesions that were visible on ultrasound. METHODS: Fifty ultrasound-guided mammotome biopsies were performed. All were small breast lesions primarily detected by ultrasound. All received FNAC as initial assessment. Mammotome biopsy was performed whenever the breast lesion was considered indeterminate or if it was considered benign and there were associated risk factors such as a family history of breast cancer. RESULTS: Of 50 mammotome biopsies 45 had benign histology. Three of 45 lesions were excised at the patients' request and were confirmed to be benign. The remaining 42 patients received an ultrasound follow up at 6 months. The lesion size remained static in 39 patients. In three patients the lesion size increased and they were excised and histology was benign. For the four malignancies diagnosed with mammotome biopsy, three patients received definitive treatment and one patient defaulted. There was one failed mammotome biopsy in the present series. CONCLUSIONS: Mammotome biopsy is an acceptable diagnostic method for small breast lesions seen on ultrasound. It reduces the need for open biopsy without compromising diagnostic accuracy. PMID- 11504289 TI - An epidemiological view of vascular trauma in Western Australia: a 5-year study. AB - BACKGROUND: Although Australia encounters vascular injury less frequently than countries such as the USA, the predominance of blunt trauma with associated complex injuries continues to pose problems for clinicians. The present paper reviews Royal Perth Hospital's experience of vascular trauma epidemiology. METHODS: All individuals who presented to the Royal Perth Hospital (RPH) with a vascular injury between August 1994 and January 2000 were identified from a prospective trauma database. Injuries were classified using the Abbreviated Injury Score (AIS 90 code). The five major vascular injury groups as defined by AIS 90 (neck, thorax, abdomen, upper limb and lower limb) were analysed in turn to determine which injuries were most frequent, what the common causes were, effects of alcohol and drugs, common associated injuries, investigations performed, mortality, proportion of rural vascular trauma with transfer details and lengths of hospital stay. RESULTS: During the study period, 153 patients (1% of total trauma workload) sustained 175 vascular injuries. The commonest causes of injury for each group were stabbing (neck and lower limb), motor vehicle accident (thorax and abdomen) and work machinery (upper limb). Mortality was highest for thoracic injury (14/32; 44%) followed by abdominal injury (10/29; 34%). The ratio of blunt to penetrating vascular trauma is high and is increasing, as is the ratio of truncal to extremity vascular trauma. The most common artery-vein combination injury was to the femoral vessels. CONCLUSIONS: The injury patterns emerging from the present study will hopefully help all medical personnel to recognize the potential for vascular injury in a trauma setting. PMID- 11504290 TI - Human bites of the hand: the Tasmanian experience. AB - INTRODUCTION: The purpose of the present paper is to provide geographically representative information on the presentation, treatment and complications of human bites of the hand in Australia. METHODS: A 5-year retrospective study was undertaken of patients treated at Royal Hobart Hospital for human bites of the hand. The variables evaluated included age, sex, hand involvement, anatomical distribution, presentation, operative findings, bacteriology, antibiotic use, X ray findings, complications and compliance. RESULTS: Thirty-five patients had human bites to the hand. All were male with a median age of 24 years. Most were clenched fist injuries with the middle metacarpophalyngeal joint being the most prone to injury. Patients presenting early had a high incidence of open joints and fractures but spent less time in hospital than late presenters, almost all who had infection complications. Compliance with treatment was found to be a major problem. Serious hand infections were not observed. CONCLUSION: The treatment of human bites must be early, correct and comprehensive. The appropriate treatment is surgical exploration with debridement and lavage, appropriate antibiotic administration, hand elevation and initial immobilization. Poor compliance of patients in the present study was demonstrated by a high incidence of late presentation, self-discharge from hospital and loss to follow up, making assessment of outcomes difficult. Despite this there is an absence of serious hand infections reported in the literature and this may be due to the administration of antibiotics prior to referral. PMID- 11504291 TI - Comparison of hydrogen peroxide instillation with Goodsall's rule for fistula-in ano. AB - BACKGROUND: Identification of the internal opening is an essential step in the management of fistula-in-ano. The predictive accuracy of Goodsall's rule is compared with instillation of hydrogen peroxide for fistulas-in-ano. METHODS: Thirty-five patients (32 male, three female; median age 42 years; range: 6 months 70 years) were studied. Hydrogen peroxide solution was injected into the external opening of the fistula track and effervescence was observed at the internal opening within the anal canal. A fistula track was either excised or incised. Setons were placed within high fistulas. RESULTS: There were 24 simple fistulas, compared to 11 complex fistulas (horseshoe, n = 4; abscess, n = 4). Eighteen external openings were anterior and 17 were posterior. Thirty-four of 35 (97%) internal openings were identified. Only 20 internal openings were in accordance with Goodsall's rule (positive predictive value: 59%). Predictive accuracy was greater for anterior external openings (13 of 18 (72%)) versus posterior external openings (six of 17 (41%); P = 0.016). For recurrent fistulas, seven of 17 fistula tracks had an internal opening in accordance with Goodsall's rule, resulting in a positive predictive value of 41%. (Positive predictive value: anterior 67% vs posterior 12.5%; P = 0.0009.) CONCLUSION: The overall predictive accuracy of Goodsall's rule was poor chiefly because of poor predictive accuracy in posterior and recurrent fistulas. The use of Goodsall's rule alone in decision making before surgical intervention is not recommended. PMID- 11504292 TI - Endocrine tumours of the pancreas: review and recent advances. AB - Pancreatic endocrine tumours (PET) are rare but nonetheless important to recognize and treat in a timely fashion. Significant morbidity occurs due to excess secretion of hormones, with all of the PET having some degree of malignant potential. Surgeons must plan directed operative strategies to deal with these tumours and be prepared to undertake aggressive palliative debulking resections if indicated. Somatostatin receptor scintigraphy and endoscopic ultrasound have been particularly helpful in both localizing and staging patients with PET. Other important advances in management include the use of long-acting somatostatin analogues to inhibit hormonal secretion and tumour growth. The possibility of multiple endocrine neoplasia type 1 (MEN-1) should be considered in any patient with a PET. The present article will review the various classes of PET, describe MEN-1 in relation to PET and examine advances in imaging and localization. The role of surgery for PET is also discussed in the present review. PMID- 11504293 TI - A simple technique for closure of the last laparoscopic working port. AB - Closure of the last laparoscopic working port can be frustrating, particularly in the very obese. A technique is described that simplifies this procedure, using a grasping forcep to transfer the abdominal wall suture. PMID- 11504294 TI - Intraoperative vascular localization to facilitate endopyelotomy after renal transplantation. AB - BACKGROUND: Pelviureteric junction (PUJ) obstruction after renal transplantation is uncommon. Surgical correction can be technically challenging due to dense perinephric adhesions and variable hilar vascular anatomy. Endopyelotomy is well established in the treatment of PUJ obstruction in native kidneys. METHODS: The present paper reports the first experience of antegrade visual cold-knife endopyelotmy performed in a renal allograft. In orientating the incision at the PUJ, preoperative imaging was supplemented by intrarenal Doppler ultrasound, using a probe designed for transoesophageal cardiac monitoring. To the authors' knowledge this approach has not previously been reported. RESULTS: Renal vascular relationships were readily indentified by identifying arterial and venous waveforms. CONCLUSIONS: For this uncommon procedure the use of intrarenal Doppler ultrasound provides greater security in avoiding inadvertent vascular injury. PMID- 11504295 TI - Island flap perineoplasty for coverage of perineal skin defects after repair of cloacal deformity. AB - BACKGROUND: Traumatic cloacal defect is an injury sustained during childbirth in which the anovaginal septum is completely disrupted and the anus and vagina open as a common channel. Such injuries result in complete faecal incontinence and are difficult to repair both in terms of improving function and obtaining skin closure. METHODS: Four cases of traumatic cloacal defect with a delayed presentation are illustrated here. All were treated with an overlapping anterior sphincter repair in combination with island flap perineoplasty to achieve skin closure. Anorectal function before and after surgery and the success of achieving primary wound healing were evaluated. RESULTS: In all four cases profound incontinence was found preoperatively; all patients returned to normal or near normal continence following overlapping anterior sphincter repair. Island flap perineoplasty was successful in achieving primary healing in all cases and no flaps were lost to necrosis. CONCLUSIONS: Island flap perineoplasty is an effective method of achieving skin closure after anterior sphincter repair of traumatic cloacal defects. PMID- 11504296 TI - Osteoid osteoma in a young child: successful non-operative management. PMID- 11504297 TI - Endoscopic management of a non-Zenker's diverticulum. PMID- 11504298 TI - Ectopic pituitary adenoma in the sphenoid sinus causing acromegaly associated with empty sella. PMID- 11504299 TI - Surgical repair of primary infundibular stenosis in a 72-year-old man. PMID- 11504300 TI - Fatal Clostridium difficile infection of the small bowel after complex colorectal surgery. PMID- 11504301 TI - Genetic Epidemiology--science and ethics on familial cancers. AB - Genetic epidemiology provides data on cancer etiology, familial risks and genotype-specific risks. These data are useful for clinical counselling and gene identification. The studies require large, unbiased sample sizes and collaboration between research teams, nationally and internationally. A recent study on Nordic twins suggests that in colorectal, breast and prostate cancer, the inherited component ranges between 27 and 42%, far in excess of the known susceptibility genes. The data from the Swedish Family-Cancer Database, particularly on second cancers, also suggest that a main genetic component in cancer is polygenic. The results have implications for design of genetic studies and for clinical counselling. PMID- 11504302 TI - Risk in numbers--difficulties in the transformation of genetic knowledge from research to people--the case of hereditary cancer. AB - Difficulties in communicating diagnostic information are exacerbated when the 'diagnosis' is a 'genetic risk' for cancer. The risk estimation demanded in this situation differs from other types of probability estimations. Observations of participants in 45 consultation sessions between physicians and potential patients were conducted at a clinic for hereditary cancer to explore the communication of genetic information. Thirty-three sessions were audiotaped, transcribed verbatim and analyzed, along with notes from the other sessions. A dominant theme was found to be numerical discussion of risk. Further analysis resulted in the description of problems for practitioners in the process of translating scientific knowledge into clinical management. Problems in providing information include unclear aims of the consultation sessions, mixing various types of background information and probabilities, recognizing how low the predictive values are, and difficulties in communicating the relationship between probability and conclusions. Problems in communicating information about the genetic risk for cancer are of at least two types: dilemmas arising from uncertainties implicit in the nature of the information itself and difficulties in communicating information in a manner that those concerned can interpret. These issues need clarification, so that information with far-reaching consequences can be made as clear and comprehensible as possible for those involved. PMID- 11504303 TI - Satisfaction with routine follow-up visits to the physician--the needs of patients with breast cancer. AB - Follow-up visits after surgery for breast cancer constitute a large proportion of the outpatient consultations at surgical and oncological clinics. The reasons for the follow-ups include early detection of relapse, patients' well-being, and data collection for quality assurance and scientific studies. The aim of this study was to describe the needs of the patient with breast cancer and satisfaction with routine follow-up visits to the physician. A strategic sample of 20 women with breast cancer, routinely followed-up at an oncology outpatient clinic, was interviewed. A qualitative descriptive design inspired by the phenomenographic method was used. The results identified the need for routines, accessibility, security, continuity, confidence and information. The women's views demonstrated that there are strong reasons for reviewing and changing the design of the traditional follow-up system to obtain the most effective and well-functioning system possible to better meet these women's needs. PMID- 11504304 TI - Mammographic parenchymal patterns and breast cancer natural history--a case control study. AB - The relationship between tumour size, malignancy grade and dense mammographic parenchymal patterns was evaluated by using a nested case-control design with 875 breast cancer cases and 2 601 matched controls. Wolfe's classification was used to assess mammographic parenchymal patterns. The dense P2/DY mammographic parenchymal patterns were significantly associated with invasive ductal grade 3 carcinoma (OR = 2.03; 95%, CI 1.13-3.62, p = 0.016). Stratified by tumour size, we found that the odds ratio for grade 3 invasive ductal NOS breast cancer measuring 15-19 mm associated with P2/DY mammographic parenchymal patterns was 3.46 (95% CI 1.12-10.68). For tumours larger than 30 mm, the odds ratio was 10.09 (95% CI 1.27-79.93). The highest risk of grade 3 cancers being in tumours measuring 30 mm + may be due to dedifferentiation of missed cancers, but there is some excess even in the < 20 mm tumours, suggesting an increased risk in association with dense mammographic patterns of some aggressive cancers which are grade 3 at inception. Our results are consistent with the model that breast cancer is a progressive disease, whose development can be arrested by screening, and that the point at which individual tumour progression is stopped is crucial for prognosis not simply in terms of stage of disease, but also of histological grade. PMID- 11504305 TI - Risk factors for male breast cancer--a case-control study from Scandinavia. AB - We report a population-based case-control study on risk factors for male breast cancer. Data on a broad range of previously suggested risk factors were collected in a set of Scandinavian breast cancer cases and matched controls. Incident cases (n = 282) with histologically verified carcinomas of the breast were identified from notification to the cancer registries of Denmark, Norway and Sweden over a 4 year period 1987-1991 and of these cases, 156 men could be approached and responded. Controls were identified through national central population registers and were matched individually for country, sex and year of birth. Controls with a diagnosis of breast cancer were excluded; 468 of 780 controls responded. Data on risk factors were collected by self-administered questionnaires mailed to the cases between land 2 years after diagnosis and to controls during the same period. The findings were compatible with an increased risk associated with family history of breast cancer (odds ratio (OR) = 3.3, 95% confidence interval (CI) 2.0-5.6), obesity 10 years before diagnosis (OR = 2.1, 95% CI 1.0-4.5) for BMI > 30, diabetes (OR = 2.6, 95% CI 1.3-5.3) and the use of digoxin and methyldopa (OR = 2.0 and 2.1, respectively). The association with family history of breast cancer has been repeated in several studies, while the relation to anthropometric measures has been equivocal. We could not substantiate some associations seen in other studies; namely those with high education, fertility, marital status, testicular injury, liver disease and religion. The detailed questions about gynaecomastia indicated that many cases reported signs of breast cancer as a gynaecomastia. This type of misunderstanding may explain the strong association with gynaecomastia seen in other studies. Several patients died before contact. Thus, risk factors related to a more aggressive male breast cancer or related to high risk of dying (e.g. liver cirrhosis, heavy smoking) may have been missed. PMID- 11504306 TI - Male breast cancer--a reappraisal of clinical and biologic indicators of prognosis. AB - Between 1970 and 1998, 90 cases of male breast cancer with available pathological material were retrieved. The disease often presented in aged patients (median--66 years) and as advanced stage (stage III/IV-51%). Excluding stage IV disease, the neoplasia were predominantly ductal invasive carcinomas. NOS (not otherwise specified) (92%), grade 1 and grade 2 (94%), positive for estrogen and progesterone receptors (72% and 74%), negative for androgen receptors (100%), p53 negative (95%), c-erbB-2 negative (88%) and DNA aneuploid (73%). Assessment of disease outcome is determined by stage at time of diagnosis, and axillary lymph node status was the only parameter found to have a statistically significant correlation with either disease-free interval or overall survival (p < 0.001) by multivariate analysis. Clinically useful information on the probability of relapse can be added by determining c-erbB-2 (p = 0.02) and progesterone receptors (p = 0.04) in stage III and tumor ploidy (p = 0.04) in pN1 subgroups of patients. PMID- 11504307 TI - Gender and time differences in initial location and histological grade of non Hodgkin's lymphomas in Southern Sweden. AB - In accordance with international data, non-Hodgkin's lymphomas (NHL) in the Southern Health Care Region of Sweden were more common in men than in women and there was an inreasing incidence for both genders in the 1980s and 1990s. A study of the years 1983 and 1992, i.e. the beginning and the end of a 10-year period, was carried out. Case records for 94% of all adults diagnosed in 1983 and 1992 (220 men and 171 women) were reviewed. The sites of the lymphomas and malignancy grade (Kiel classification) were recorded. Comparisons were made between the genders and between the two years studied. Among 170 men with nodal lymphomas, 49 were localized (stage I-II) in lymphoid tissue regional to the respiratory tract, 18 were localized elsewhere and 103 were generalized (stage III-IV). The corresponding sites in 138 women were 28, 33 and 77, respectively (p = 0.002), implying a predominance of stage I-II lymphomas regional to the respiratory tract in men and a larger number of localized lymphomas elsewhere, mostly abdominal retroperitoneal with or without inguinal engagement in women. High-grade nodal lymphomas were more common in 1992 (54%) than in 1983 (34%, p = 0.01). suggesting an increasing number of mainly high-grade lymphomas. For extranodal lymphomas (50 in men and 33 in women), no clear differences were observed for gender and the years studied. Risk factors, e.g. occupational hazards, are probably different for men and women. The gender differences in initial nodal location suggest that initial lymphoma sites may be related to different risk factors. Epidemiologic studies where both malignancy grade and initial lymphoma locations are taken into account may be helpful in defining risk factors. PMID- 11504309 TI - Prognostic value of histopathological response to radiotherapy and microvessel density in oral squamous cell carcinomas. AB - The prognostic value of histopathological response to preoperative radiotherapy (50 Gy) in radically resected oral carcinomas was studied in 39 consecutive patients. Microvessel density (MVD) was evaluated for relation to radioresponse and outcome. Resected tumour tissue was examined histopathologically and response to radiotherapy was scored according to induced morphological changes. Pretreatment biopsies were stained with antibodies to von Willebrand factor to evaluate MVD in hot-spot regions, in stromal tissue and in tumour epithelial tissue. Histopathological response to radiotherapy was highly prognostic of local failures and survival (p = 0.002), though microscopic surgical radicality was obtained. In good responders to preoperative radiotherapy, the 5-year survival rate was 68% compared with 24% in poor responders. In 12 patients with local recurrence after radical surgery, 11 had poor histopathological radiotherapy responses. In univariate analysis, a high MVD score in tumour epithelium was associated with poor clinical outcome but MVD did not correlate with histopathological radiotherapy response. PMID- 11504308 TI - Non-Hodgkin's lymphoma of the ocular adnexa. AB - This study investigates the relationship between the clinical features of lymphoma in the ocular adnexal region and the revised European and American lymphoma (REAL) classification. Specimens from 41 patients with ocular adnexal lymphoproliferative disease were reassessed pathologically using the REAL classification. Thirty-two patients with primary non-Hodgkin's lymphomas (NHL) were included in the study, almost all of them having been treated with radiotherapy with or without chemotherapy. Seven of the 32 patients with NHL showed distant recurrence after treatment: 3 out of 26 with extranodal marginal zone B-cell lymphoma, and 4 with other types of NHL. Although the three patients with recurrent marginal zone B-cell lymphomas all survived, other patients with recurrent lymphomas died of disease. The REAL classification provides a good indication of tumor control probability and survival of patients with ocular adnexal NHL. Radiation therapy is an effective treatment modality for extranodal marginal zone B-cell lymphoma of the ocular adnexa. PMID- 11504310 TI - Improving local tumor control by combining vascular targeting drugs, mild hyperthermia and radiation. AB - Improvement in local control in a foot-implanted (200 mm3) C3H mouse mammary carcinoma by combining vascular targeting drugs, mild hyperthermia and radiation was investigated. The vascular targeting drug was flavone acetic acid (FAA; 150 mg/kg) intraperitoneally injected either 3 h before local tumor water-bath heating or 1 h after local tumor irradiation. For untreated tumors, the average (+/- 1 S.E.) tumor growth time (TGT; time to reach 5 x treatment volume) was 7.1 days (+/- 0.4). This was increased to 9.2 days (+/- 0.7) by using FAA. Heating also increased TGT, the effect being temperature and time dependent, and this heat response was further increased by FAA. The radiation dose (+/- 95% confidence interval) to control 50% of tumors (TCD50) 90 days after irradiation was 52 Gy (50-55) for radiation alone. This was decreased to 42 Gy (39-45) by FAA, 47 Gy (45-50) by heating (41.5 degrees C; 60 min) 4 h after irradiation, and to 28 Gy (22-35) by combining FAA and heat. Thus, vascular targeting drugs can improve the efficacy of mild hyperthermia and radiation. PMID- 11504311 TI - Cyclin D1 overexpression versus response to induction chemotherapy in squamous cell carcinoma of the head and neck--preliminary report. AB - The aim of this study was to investigate whether there is an association between overexpression of cyclin D1 and response to therapy. Immunohistochemical overexpression of cyclin D1 was determined in paraffin-embedded specimens from diagnostic biopsies of 89 primary cases of squamous cell carcinoma of the head and neck (SCCHN), using a polyclonal antiserum. The tumor response rates were estimated after curative treatment (i.e. surgery and/or radiotherapy and/or chemotherapy). Patients whose tumors were overexpressing cyclin D1 showed complete or partial response to neoadjuvant chemotherapy with cisplatin/5-FU. In addition, a majority of cyclin D1 negative tumors did not respond at all to this treatment (p = 0.02, Fisher's exact test). This study indicates that immunohistochemical assessment of cyclin D1 expression in SCCHN could be a new predictive marker to select a subgroup of patients that will benefit from neoadjuvant chemotherapy. PMID- 11504312 TI - Biomarkers of intrinsic angiogenic and anti-angiogenic activity in patients with endometrial hyperplasia and endometrial cancer. AB - Serum vascular endothelial growth factor (VEGF) and endostatin were determined in postmenopausal women, including 72 with endometrial cancer, 27 with endometrial hyperplasia and 30 healthy controls. Serum VEGF levels in endometrial hyperplasia (142+/-18 ng/ml, mean +/- SE) and endometrial cancer stages I (291+/-22), II (623+/-68) and stage III-IV (1527+/-119) were significantly higher than the mean for controls (12+/-1.6). Serum endostatin levels in endometrial hyperplasia (149+/-19 ng/ml), endometrial cancer stages I (320+/-41), II (644+/-86) and stage III-IV (1253+/-114) were also significantly higher than the mean for controls (13+/-2.4). Elevated values of VEGF above the non-malignant level were encountered in 7% (stage I), 37% (stage II) and 100% (stage III-IV) of endometrial cancers. The corresponding figures for endostatin were 37%, 59 and 100%, respectively. These results demonstrate that the circulating levels of both markers correlated with tumor stage and apparently tumor burden. Serum VEGF and endostatin levels decreased significantly after treatment, followed by marked elevations at clinical relapse. The VEGF endostatin ratio was higher in the advanced stages ( > 1.0) than in the early stages of endometrial carcinoma (< 1.0). indicating that the balance of angiogenic stimulators and inhibitors may regulate metastasis and access tumor progression. PMID- 11504313 TI - Radiobiological hypoxia, oxygen tension, interstitial fluid pressure and relative viable tumour area in two human squamous cell carcinomas in nude mice during fractionated radiotherapy. AB - Very little is known about the correlation between the radiobiological hypoxic fraction (rHF) and other measures of tumour oxygenation during fractionated irradiation. In the present study the rHF is determined in untreated human FaDu and GL squamous cell carcinoma in nude mice and in tumours irradiated with 10 fractions in 2 weeks and 20 fractions in 4 weeks, using tumour control as the experimental endpoint. The results were compared with measurements of the pO2, the interstitial fluid pressure (IFP) and the relative viable tumour area. In FaDu tumours the radiobiological hypoxic fractions (rHFs) before and during irradiation were not statistically different from 100%. Depending on the assumptions made for D0, the rHFs of GL tumours were between 0.2 and 4% or 30 and 53%. The median pO2 values were 2.8 mmHg for untreated FaDu tumours and 0.2 mmHg for GL tumours (p < 0.001). The median IFP values were 2.6 mmHg in FaDu and 5.3 mmHg in GL tumours (p = 0.01). No important changes in the pO2 and IFP values were observed during fractionated irradiation. The relative viable tumour area during irradiation decreased by 83% in FaDu tumours (p = 0.002) and by 54% in GL tumours (p = 0.003). It is concluded that differences in rHF exist between FaDu and GL tumours before and during fractionated irradiation and that these differences are not reflected by pO2 and IFP values and the relative viable tumour area. PMID- 11504315 TI - Serum lactate dehydrogenase isoenzyme 1 and relapse in patients with nonseminomatous testicular germ cell tumors clinical stage I. AB - Serum lactate dehydrogenase isoenzyme 1 catalytic concentration (S-LD-1) was measured at the time of orchiectomy in 104 patients with nonseminomatous testicular germ cell tumors (NSTGCT) clinical stage I who participated in a randomized study comparing surveillance after orchiectomy (group I) and radiotherapy (group II). For 68 patients, S-LD-1 was measured in a serum sample before or on the day of the orchiectomy. Twenty-seven patients (40%) had elevated S-LD-1; median 102 U/L (range 41-335). For the remaining 36 patients. S-LD-1 was measured in a serum sample after orchiectomy: 8 of these patients (22%) had elevated S-LD-1. S-LD-1 was normalized shortly after surgery in most patients with a preorchiectomy elevated S-LD-1. Fifteen of the 68 patients relapsed: 9 out of 27 with an elevated S-LD-1 and 6 out of 41 patients with normal S-LD-1 (p = 0.13, Fisher's exact test). In group 1, those with a preoperatively elevated S-LD 1 had a lower 8-years' relapse-free survival than those with a normal S-LD-1 (40% vs. 80%, p = 0.003, log-rank test). The role of S-LD-1 in the staging, prognostication and monitoring of patients with NSGCT clinical stage I should be further explored in a large, prospective study. PMID- 11504314 TI - Gamma-glutamyl transferase expression in higher-grade astrocytic glioma. AB - Increased expression of gamma-glutamyltransferase (GGT) has been detected in a range of human malignancies and is thought to be involved in neoplastic proliferation and treatment resistance. Since GGT expression and its role in malignant glioma biology remain largely unknown, we investigated this phenomenon by immunostaining 26 higher-grade human astrocytic gliomas (WHO grades III and IV) with a monoclonal anti-GGT-antibody (138H11). Further, human pancreatic GGT cDNA was used for liposome-mediated transfection of 9L gliosarcoma cells. GGT expressing and control 9L cells were cultured in media containing different amounts of essential amino acids and/or cytotoxic agents. Cell viability was evaluated by microplate MTT assay. Immunohistochemical staining of tumor specimens demonstrated that GGT expression is a frequent feature of higher-grade human astrocytic gliomas, but not of normal brain tissue. Human tumors were strongly GGT-positive in 6 of 7 cases of grade III astrocytoma, and in 12 of 19 grade IV astrocytoma (glioblastoma multiforme, GBM) cases. In the cell culture model, 9L-GGT cells had a growth advantage over control cells in cysteine deficient medium. but not in standard or glutamine-free medium. No significant difference in numbers of viable cells of either clone was found in media containing the alkylating drug BCNU (5-200 microg/ml). In conclusion, GGT is expressed in a high percentage of human WHO grade III astrocytomas and GBM, but not in normal brain tissue. This molecule seems to give neoplastic cells a moderate growth advantage under in vivo conditions. PMID- 11504316 TI - A new conservative approach to extravasation of anthracyclines with dimethylsulfoxide and dexrazoxane. PMID- 11504317 TI - A population-based twin study of generalized anxiety disorder in men and women. AB - This study aimed to a) assess whether genetic or environmental effects are of similar magnitude in the etiology of GAD in men and women, and b) investigate whether familial (genetic or common environmental) risk factors are the same in men and women, or whether there are gender-specific effects. We obtained a lifetime history of DSM-IIII-R GAD, via face-to-face and telephone interviews, from 3100 complete male-male, female-female, and male-female twin pairs, ascertained through a population-based registry. Biometrical twin modeling was utilized to estimate the relative contributions of genetic and environmental factors to liability for GAD, allowing for gender-specific effects. The familial aggregation of GAD in this sample was only modest. In the best-fitting models, the heritability of GAD was the same in men and women, estimated at about 15% to 20%, with no effects of gender-specific genes detected. PMID- 11504318 TI - The effect of remission of poststroke depression on activities of daily living in a double-blind randomized treatment study. AB - Poststroke depression has been associated with impaired recovery of activities of daily living (ADL) during the first 2 years after stroke. This study examined the effect of remission of poststroke depression on recovery in ADL in a double-blind randomized treatment study. Based on a semistructured psychiatric exam and DSM-IV diagnostic criteria, a consecutive series of 23 patients who met criteria for major depression (N = 16) or minor depression (N = 7) were selected and randomly assigned to either active treatment (nortriptyline) or placebo. Functional physical (i.e., ADL) impairment was assessed using the Johns Hopkins Functioning Inventory (JHFI). Patients whose depressive disorder remitted at follow-up had significantly greater recovery in ADL functions compared with patients whose depression did not remit. There were no differences in demographic variables, lesion characteristics, and neurological symptoms between the two groups, which would explain the significantly greater improvement among the remitted patients. Because both major and minor depression patients who remitted showed greater improvement in ADL than nonremitted patients some of whom were treated with active and some with placebo medication, nonpharmacotherapeutic mechanisms related to recovery from depression appear to mediate this enhanced recovery. PMID- 11504319 TI - Function after motor vehicle accidents: a prospective study of mild head injury and posttraumatic stress. AB - Relationships among mild traumatic brain injury (MTBI), posttraumatic stress (PTS), and function were examined in 99 motor vehicle accident (MVA) admissions: 64 in an MTBI group and 35 in a no-MTBI comparison group. Assessments occurred within the first month and at 6 to 9 months. At follow-up, the sample was moderately disabled on the Sickness Impact Profile (SIP), 71% satisfied on the Reintegration of Normal Living Index (RNL), and 42% had returned to work. Only the SIP Psychosocial score was significantly different for MTBI groups; 24% of the sample showed definite symptoms of PTS. This group was significantly more disabled on the SIP, less satisfied on the RNL, and less likely to return to work. The proportion of variance in outcome explained in each model ranged from 32% (Physical SIP) to 44% (RNL). Results suggest the need for clinicians to be more aware of the strong influence of PTS on functional outcomes. PMID- 11504320 TI - Predictive validity of the physical disorders axis of the DSM multiaxial diagnostic system. AB - This paper reports on the predictive validity of the physical disorders axis (axis III) of the DSM multiaxial diagnostic system at 3-year follow-up. A total of 515 general psychiatric patients were assessed with a semistructured procedure that covers all DSM-III diagnoses and axes, and were subsequently followed up for 3 years. Outcome was assessed with several measures of adaptive functioning. Baseline axis III was analyzed according to a) presence of any physical disorder, b) the number of these, c) presence of major chronic physical disorders (MCPD), and d) the number of these. Prediction of impairment in functioning (Strauss Carpenter Scale), derived from baseline axis III, ranged from a correlation coefficient of .18 when expressed as the presence of any physical disorder to .35 when represented by the number of MCPD. Furthermore, within patients with specific psychiatric disorders, it was found that number of MCPD reached a predictive validity of .55 for patients with dysthymic disorders, .44 for those with anxiety disorders, and .41 for those with major depression. Comparative multiple regression analyses, controlling for demographic and clinical variables, showed that the number of MCPD at baseline was the most important predictor of functioning outcome among patients with dysthymic disorders and major depression. The number of MCPD experienced by general psychiatric patients seems to be an important predictor of future functioning, particularly for patients with certain psychiatric disorders. This points out the importance of considering the relationship between psychiatric and MCPD when conducting systematic clinical assessments towards the prediction of course and outcome. PMID- 11504321 TI - Effects of gender and ethnicity on duty-related posttraumatic stress symptoms among urban police officers. AB - We studied 655 urban police officers (21% female, 48% white, 24% black, and 28% Hispanic) to assess ethnic and gender differences in duty-related symptoms of posttraumatic stress disorder (PTSD). We obtained self-report measures of: a) PTSD symptoms, b) peritraumatic dissociation, c) exposure to duty-related critical incidents, d) general psychiatric symptoms, e) response bias due to social desirability, and f) demographic variables. We found that self-identified Hispanic-American officers evidenced greater PTSD symptoms than both self identified European-American and self-identified African-American officers. These effects were small in size but they persisted even after controlling for differences in other relevant variables. Contrary to expectation, we found no gender differences in PTSD symptoms. Our findings are of note because: a) they replicate a previous finding of greater PTSD among Hispanic-American military personnel and b) they fail to replicate the well-established finding of greater PTSD symptoms among civilian women. PMID- 11504322 TI - Gender differences in the relationship of homelessness to symptom severity, substance abuse, and neuroleptic noncompliance in schizophrenia. AB - This study examined gender differences in the relationship of homelessness in schizophrenia to symptom severity, risk behaviors, and prognostic features. Four hundred subjects with schizophrenia were studied: 100 homeless men, 100 homeless women, 100 never homeless men, and 100 never homeless women. Assessments included derivation of five symptom factors by using the Positive and Negative Syndrome Scale (PANSS). Homelessness for the entire sample was associated with greater severity of positive, activation, and autistic preoccupation symptoms, younger age at first hospitalization, and substance abuse (SA). For men only, homelessness was associated with neuroleptic noncompliance (NN). When NN and SA were statistically controlled, symptom severity was not different between the homeless and never homeless. Women, independent of residential status, had more severe negative, activation, and autistic preoccupation symptoms that were not associated with prognostic features or risk behaviors. For both men and women, SA was associated with homelessness, but independent of residence, SA was less severe in women. Additionally, SA was less severe in homeless women than never homeless men. Thus, symptom severity in homeless individuals with schizophrenia appears as an interaction of symptom profiles and risk behaviors that are gender specific. Although cross-sectional analyses cannot distinguish cause from effect, these findings suggest gender-specific routes to homelessness among indigent urban adults with schizophrenia. PMID- 11504323 TI - Neurocognitive correlates of helplessness, hopelessness, and well-being in schizophrenia. AB - Persons with schizophrenia are widely recognized to experience potent feelings of hopelessness, helplessness, and a fragile sense of well-being. Although these subjective experiences have been linked to positive symptoms, little is known about their relationship to neurocognition. Accordingly, this study examined the relationship of self-reports of hope, self-efficacy, and well-being to measures of neurocognition, symptoms, and coping among 49 persons with schizophrenia or schizoaffective disorder. Results suggest that poorer executive function, verbal memory, and a greater reliance on escape avoidance as a coping mechanism predicted significantly higher levels of hope and well being with multiple regressions accounting for 34% and 20% of the variance (p < .0001), respectively. Self-efficacy predicted lower levels of positive symptoms and greater preference for escape avoidance as a coping mechanism with a multiple repression accounting for 9% of the variance (p < .05). Results may suggest that higher levels of neurocognitive impairment and an avoidant coping style may shield some with schizophrenia from painful subjective experiences. Theoretical and practical implications for rehabilitation are discussed. PMID- 11504324 TI - Social phobia as a comorbid condition in sex offenders with paraphilia or impulse control disorder. AB - Studies on the prevalence of social anxiety in sex offenders show mixed results. This may be due to social anxiety being heightened only in diagnostic subgroups of sex offenders, namely in paraphiliacs. In study 1, 72 mentally disordered sexual delinquents and 30 controls were screened for social anxiety with the Social Interaction Anxiety Scale and the Social Phobia Scale by Mattick and Clarke (German versions). In study 2, 55 mentally disordered sexual delinquents were diagnosed with a structured clinical interview. In both studies, sex offenders were categorized as either paraphilic or impulse control disordered (without paraphilia) according to research criteria. Study 1 showed markedly heightened scores for social anxiety in paraphiliacs, particularly for social interaction anxiety. Study 2 found a high lifetime and point prevalence of social phobia in paraphiliacs for which corroborating evidence was again found in questionnaire results. Implications for further research, diagnostic procedures, and therapy are discussed. PMID- 11504325 TI - Developmental aspects of obsessive compulsive disorder: findings in children, adolescents, and adults. AB - Although juvenile obsessive compulsive disorder (OCD) is increasingly recognized as a putative developmental subtype of the disorder, comparisons among children, adolescents, and adults with OCD have been lacking. We aimed to evaluate clinical correlates of OCD in three developmentally distinct groups. Subjects comprised children, adolescents, and adults meeting DSM-III-R and DSM-IV criteria for OCD referred to separate specialized OCD clinics. All subjects were systematically evaluated with structured diagnostic interviews and clinical assessments by OCD experts. Specific clinical correlates and symptom profiles were associated with the disorder in different age groups. These findings support a hypothesis of developmental discontinuity between juvenile and adult OCD and identify age specific correlates of the disorder across the life cycle. Further work is needed to validate whether juvenile-onset OCD represents a true developmental subtype of the disorder. PMID- 11504326 TI - When dreams become a royal road to confusion: realistic dreams, dissociation, and fantasy proneness. AB - Scientific discussions about false memories have, so far, mainly focused on external determinants (e.g., therapeutic interventions). However, in some cases, false memories might develop more spontaneously. For example, difficulties in distinguishing between dreams and reality may lead to false memories. The present article discusses two studies (N = 85 and 255, respectively) that examined to what extent such difficulties occur. In both studies, a nontrivial minority of respondents (11.8% and 25.9%, respectively) reported that they had had the experience of not being able to discriminate between dream and reality. As expected, respondents who reported this type of confusion scored higher on fantasy proneness and dissociation measures than respondents who did not report this confusion. PMID- 11504328 TI - Biochips 2001: the second-generation chip for the clinic. PMID- 11504327 TI - The limited influence of latitude on rates of seasonal affective disorder. PMID- 11504329 TI - Point mutation detection technology for the new-century laboratory. AB - The READIT system represents the newest contribution to point mutation detection technology. Specifically, the system involves hybridizing DNA or RNA probes using phosphorylation chemistry and luciferase as the detection method. The two primary features that distinguish the READIT technology from the competition are its versatility and the ability of users to design its own probes. The automated genotype assignment and statistical analysis simplify the use of this system even more. In the year 2002 more than a million tests will be performed in molecular diagnostics with significant contributions from both research and private laboratories.(1) The most important attributes for laboratorians when reviewing a new technology are cost, accuracy, automation, and ease of use. The advent of unique point mutation detection technologies will certainly assist the role of the laboratory in demonstrating the positive impact of testing on future applications of these technologies. Cost savings, accuracy, simplicity, and flexible throughput are the must-have features of successful point mutation detection technologies for the new-century laboratory. PMID- 11504330 TI - Pathogenesis and symptoms of zinc deficiency. PMID- 11504331 TI - Resistance to protease inhibitors in HIV patients experiencing virologic failure despite adherence to antiretroviral therapy. PMID- 11504332 TI - Significantly reduced variable costs, mortality, and length of stay with timely microbiologic procedures. PMID- 11504333 TI - The perils of nonprospectively planned research. Part 1: drawing conclulsions from sample-based research. PMID- 11504334 TI - A critical appraisal of cystic fibrosis sweat-testing guidelines. PMID- 11504335 TI - ASCP/CAP 2000: keynoting 21st-century pathology informatics and integration into the emerging hospital information system. PMID- 11504336 TI - Update on HIV patient management: latest trends in laboratory testing, immune cell reconstitution, and vaccine development. AB - HIV patient management requires the close interaction of clinicians and laboratorians to integrate the results obtained by emerging methodologies. Evaluation of drug susceptibility to screen for specific mutations in the HIV genome can avoid needless treatment with ineffective drugs, maximizing the benefits of costly triple-drug therapy. In addition, providing a scientific basis for the effective use of genotyping and therapeutic drug monitoring is a valuable role that laboratorians can play in optimizing patient care. The clinical utility of HIV genotyping has been defined more clearly during the last year, and pharmacokinetic profiles of the antiretrovirals are beginning to emerge in the clinical laboratory as a tool to periodically assess adherence to and efficacy of the drug regimens. However, much more work is required in this area. Switching to simpler regimens is perceived as a means to improve medication adherence, manage drug toxicity, and reduce the potential for interaction among different drugs. HIV affects the function of both cytotoxic T-cells and neutralizing antibodies. The ability to identify and quantify HIV-specific immunity may allow us to target and expand specific deficient cell clones as a means of correcting immune deficiencies and strengthening the effects of drug therapy. One such example is IL-2, shown to increase CD4 cell counts at various stages of HIV disease. Additional studies of IL-2 therapy and newer immune modulators are ongoing. Combination vaccines are being tested as a worthwhile approach to optimize induction of both CTLs and antibodies. The role of the laboratory to determine the potency of HIV vaccine candidates awaits further developments evaluating the relative importance of neutralizing and mucosal antibodies versus CTL responses and the degree of immunogenicity needed to contain the spread of infection. The Center for Disease Control and Prevention aims to reduce new HIV infections by 50% per year in the next five years, and this goal is to be achieved through selected strategies put together by the plethora of experts in this arena. The aim of this article is to update the reader on the latest trends in HIV patient management and motivate collaborations to assist in the prompt implementation of improved medical care. PMID- 11504337 TI - Elemental concentrations in different species of seaweeds from Loreto Bay, Baja California Sur, Mexico: implications for the geochemical control of metals in algal tissue. AB - Concentration levels of 21 elements were analyzed by instrumental neutron activation analysis (Rb, Cs, Ca, Sr, Ba, Sc, Cr, Fe, Co, Ni, Zn, Se, As, Sb, Th, U, Br, Hf, Ta, Zr, and Ag) in seven different seaweed species (Codium cuneatum, Sargassum sinicola, Padina durvillaei, Laurencia johnstonii, L. papillosa, Gracilaria pachidermatica and Hypnea pannosa), collected in a shallow coastal zone from Bahia de Loreto, Baja California Sur, Mexico. Measured concentrations in algal tissue spanned almost eight orders of magnitude (from 2.0 x 10(-3) microg g(-1) for Hf to 1.2 x 10(5) microg g(-1) for Ca). Ca was consistently the most abundant element in all analyzed seaweeds, followed by Fe and Sr. Brown algae showed a tendency to incorporate higher concentrations of elements than red and green algae. Additionally, there were significant linear correlations (P < 0.05 to P < 0.001) between a total of 76 different pairs of elements, some of them (e.g. Rb-Ni, Rb-Ag, Sc Cr, Sc-Fe, Sc Ni, Sc Hf, Cr Fe, Fe Ni, Fe-Hf and Ni Th) highly correlated (r2 > 0.900). A significant correlation (r2 = 0.701, n = 18, P < 0.001) exists between our measurements in the tissue of algae and their corresponding average elemental concentrations in oceanic water from the North Pacific Ocean. Hence, overall elemental abundance in algal tissue apparently is controlled by the elemental abundance in oceanic water, whereas metabolic processes as well as environmental factors relevant to each region modify the final concentration of a given element in the body of a macroalgae. PMID- 11504338 TI - Flue gas desulfurization by-products additions to acid soil: alfalfa productivity and environmental quality. AB - Flue gas desulfurization (FGD) by-products are created when coal is burned and SO2 is removed from the flue gases. These FGD by-products are often alkaline and contain many plant nutrients. Land application of FGD by-products is encouraged but little information is available related to plant responses and environmental impacts concerning such use. Agricultural lime (ag-lime) and several new types of FGD by-products which contain either vermiculite or perlite were applied at 0, 0.5, 1.0, and 2.0 times the soil's lime requirement (LR) rate to an acidic soil (Wooster silt loam). The highest FGD by-products application rate was equivalent to 75.2 Mg ha(-1). Growth of alfalfa (Medicago sativa L.) was significantly increased compared to the untreated control in the second year after treatment with yields for the 1 x LR rate of FGD approximately 7-8 times greater compared to the untreated control and 30% greater than for the commercial ag-lime. Concentrations of Mo in alfalfa were significantly increased by FGD by-products application, compared to the untreated control, while compared to the ag-lime treatment, concentrations of B increased and Ba decreased. No soil contamination problems were observed, even at the 2xLR rate, indicating these materials can be safely applied to agricultural soils. PMID- 11504340 TI - Methane emissions from wastewater management. AB - Greenhouse gas emissions in the form of methane and carbon dioxide are produced when municipal and industrial wastewater and their residual solid by-product (sludge) are handled under or subject to anaerobic conditions, thus contributing to the global warming potential or the greenhouse effect. This paper presents estimation methods used for determining methane emissions from the management of wastewater. Applications for estimating countrywide methane gas emissions from wastewater management are presented with the country of Lebanon as an example. The relative significance of these emissions is assessed in comparison with methane emissions from developing and developed countries. Uncertainty associated with the estimation process and mitigation measures to reduce potential impacts of methane emissions from wastewater management are also discussed. PMID- 11504339 TI - Environmental cadium exposure and metabolic responses of the Nile tilapia, Oreochromis niloticus. AB - The contamination of water by metal compounds is a worldwide environmental problem. This study was undertaken to evaluate the impact of short-term cadmium exposure on metabolic patterns of the freshwater fish Oreochromis niloticus. The fish were exposed to 320, 640, 1,280 and 2,560 microg/l sublethal concentrations of Cd++ (CdCl2) in water for 7 days. The specific activities of the enzymes phosphofructo kinase (PFK-E.C.2.7.1.11.), lactate dehydrogenase (LDH E.C.1.1.1.27.) and creatine kinase (CKE.C.2.7.3.2.) were decreased in white muscle after cadmium treatments, indicating decreases in the capacity of glycolysis in this tissue. Cadmium exposure induced increased glucose concentration in white muscle of fish. On the other hand, cadmium exposure at sublethal concentrations increased phosphofructo kinase and LDH in red muscle of fish. Cadmium significantly decreased total protein concentrations in liver and white muscle regardless of tissue glycogen levels. The data suggest that cadmium acts as a stressor, leading to metabolic alterations similar to those observed in starvation. PMID- 11504341 TI - Growth and yield responses of spring wheat (Triticum aestivum L. cv. Minaret) to elevated CO2 and water limitation. AB - Spring wheat (Triticum aestivum L. cv. Minaret) was grown at two different CO2 concentrations (367 and 650 micromol mol(-1)) in open-top-chambers from sowing until final harvest. Furthermore two different watering treatments (well watered and water stressed) and two soil types of different fertility were used. At final harvest, which took place at growth stage 92, plants were separated into different fractions. Elevated atmospheric CO2 caused an accelerated chlorophyll-a breakdown and increased growth and yield. Total shoot biomass was enhanced by 43%, grain yield by 46% and main stem yield by 19%. Water stress also accelerated chlorophyll-a breakdown but reduced total shoot biomass by 40%, grain yield by 45%, main stem yield by 30% and thousand grain weight by 6%. On average, soil fertility altered shoot biomass by 30%, grain yield by 39% and main stem yield by 25%. PMID- 11504342 TI - The acute toxicity of agricultural surfactants to the tadpoles of four Australian and two exotic frogs. AB - Nonionic surfactants are frequently incorporated into pesticide formulations, and are therefore a group of chemicals to which amphibians may be exposed in agricultural or urban landscapes. However, little is known about the effects of surfactant exposure in amphibians. Feeding stage tadpoles of Bufo marinus, Xenopus laevis and four species of Australian frogs (Crinia insignifera, Heleioporus eyrei, Limnodynastes dorsalis and Litoria moorei) were exposed to nonylphenol ethoxylate (NPE) and alcohol alkoxylate in static-renewal acute toxicity tests. All species exhibited nonspecific narcosis following exposure to both these surfactants. The 48-h EC50 values for NPE ranged between 1.1 mg/l (mild narcosis) and 12.1 mg/l (full narcosis). The 48-h EC50 values for alcohol alkoxylate ranged between 5.3 mg/l (mild narcosis) and 25.4 mg/l (full narcosis). Replicate acute toxicity tests with B. narinus exposed to NPE at 30 degrees C over 96 h indicated that the narcotic effects were not particularly time dependant. The mean 24, 48, 72, and 96-h EC50 (mild narcosis) values were 3.6, 3.7, 3.5 and 3.5 mg/l, respectively. The mean 24, 48, 72 and 96-h EC50 (full narcosis) were 4.0, 4.1, 4.2 and 4.0, respectively. Acute toxicity tests with B. marinus exposed to NPE at 30 degrees C under conditions of low dissolved oxygen (0.8-2.3 mg/l) produced a two to threefold increase in toxicity. PMID- 11504343 TI - Distribution of organochlorine pesticides in sediments from Kyeonggi Bay and nearby areas, Korea. AB - The residues of oragnochlorine pesticides (OCPs) in 62 sediments from Kyeonggi Bay and nearby areas in the west coast of Korea were determined. The concentrations of chlordanes (CHLs) and DDTs showed a distinctive gradient of contamination between inner and outer sites of Incheon North Harbor (INH), whereas hexachlorocyclohexanes (HCHs) were uniformly distributed at most sites studied. The distribution of CHLs and DDTs was strongly correlated with total organic carbon contents in sediments while HCH residue levels were independent. Relationship between contaminant's concentration and environmental factors was analyzed by principal component analysis. Distribution patterns of T-CHLs, T DDTs, and TOC were similar while those of T-HCHs, mud content, and grain size were similar. The notable contamination by CHLs and DDTs was found in INH where these levels were one or two orders of magnitude higher than other sites. The dominant OCPs in sediments were beta-HCH among HCH compounds, trans-chlordane among CHL compounds, and p,p'-DDD among DDT compounds. The higher concentrations and compositional pattern of OCPs in INH sediments indicate that INH is in the vicinity of the source. PMID- 11504344 TI - A tunable diode laser system for ammonia flux measurements over multiple plots. AB - This paper reports on the field testing of a tunable diode laser trace gas analyzer system for micrometeorological monitoring of ammonia fluxes. This system uses infrared absorption spectroscopy to measure atmospheric ammonia concentrations and the fluxgradient method to relate the measured concentration gradient to a flux of ammonia. For the field tests, we monitored ammonia fuxes over three plots receiving different manure applications. Each plot was sampled for 15 or 30 min of each hour, producing a high-temporal resolution data set. Analysis of the system response showed that ammonia adsorption to the tubing walls was greatly reduced by the system design and did not interfere with the flux measurement. PMID- 11504345 TI - Response of mycorrhizal Norway spruce seedlings to various nitrogen loads and sources. AB - Norway spruce seedlings were grown under greenhouse conditions in Rootrainers with a vermiculite-peat moss mixture under various N-regimes for 6 months. Either ammonium or nitrate was applied in loads of 100 or 800 kg N ha(-1) year(-1) to seedlings which were either non-mycorrhizal or inoculated with the mycorrhizal fungi Hebeloma crustuliniforme or Laccaria bicolor. The use of increasing N loads enhanced shoot and total biomass, whereas root/shoot ratio, number of short roots and mycorrhization decreased. A significant enhancement of the concentration and content was obvious for the element N, whereas a significant decrease was obvious for P and Zn concentrations. The use of ammonium, as opposed to nitrate, significantly enhanced the biomass and the numbers of short roots, and reduced the root/shoot ratios, but did not influence the mycorrhization. It further significantly enhanced the N concentrations in roots and shoots. Fungal inoculation with H. crustuliniforme or L. bicolor compared to non-inoculated controls significantly enhanced shoot and total biomass, but reduced root/shoot ratios. The mycorrhization further significantly enhanced N and P concentrations and contents, but reduced Mn. Overall, the mycorrhization improved the P nutrition of the seedlings independently on the applied N loads or N sources. Dose response curves using ammonium nitrate as N source with a maximum load of 1600 kg N ha(-1) year(-1) applied on seedlings associated with H. crustuliniforme revealed that the maximum growth was reached at a load of 800 kg N ha(-1) year( 1) with a simultaneous decrease of the mycorrhization. In both shoots and roots, N concentrations increased constantly with increasing N loads, while P, Ca, and Zn concentrations decreased constantly. PMID- 11504346 TI - Precipitation chemistry in the coast of the Metropolitan Region of Rio de Janeiro, Brazil. AB - Precipitation chemistry was studied in the Metropolitan Region of Rio de Janeiro (MRRJ). This study reveals that rainwater in the MRRJ is affected by emissions of air pollutants and provides essential data for future estimates of regional biogeochemical cycles and the impacts of acid deposition on tropical ecosystems. The volume-weighted mean (VWM) pH was 4.77, varying from 3.50 to 6.85. Sea-salt aerosols were the dominant sources of the Na+, Cl- and Mg2+. Excess SO4(2-), Ca2+ and K+ comprised 82, 91, and 87% of their total VWM concentrations, respectively. There were very strong correlations (r > 0.75, P > 0.01) for NO3- and H+, NO3- and excess(exc-)SO4(2-), NH4+ and exc-K+, and exc-SO4(2-) and exc-Ca2+, suggesting causal relationships between these ion pairs. The VWM concentrations of all major ions, except H+, were higher in the dry season, with dry to wet VWM concentration ratios varying from 1.1 (NH4+) to 4.7 (for total K+). PMID- 11504347 TI - Temporal and biogeographic comparisons of PCBs and persistent organochlorine pollutants in the blubber of fin whales from eastern Canada in 1971-1991. AB - Concentrations of PCB congeners and organochlorine (OC) pesticides were studied in archived fin whale (Balaenoptera physalus) blubber samples collected in 1971 1972 from Newfoundland (Nfld) and Nova Scotia (NS) with the objective of obtaining a historical perspective on contaminant levels and proportions at a time when many persistent OCs were still in use. Concentrations of sigmaPCBs and sigmaDDTs in the blubber of 17 fin whales from historical whaling stations in 1971-1972 were generally in agreement with previously reported values for fin whales from Atlantic Canada. Although some differences in sex and body length (age) distribution of samples occurred, significant differences were detected in the concentrations and patterns of PCBs, DDTs and chlordanes in whales from Nfld and NS, supporting ecological evidence that whales from these regions represent relatively separate stocks. Temporal and geographical variations were examined by comparing data to those reported for fin whales from the St Lawrence Estuary (StL) in 1991 by Gauthier et al. (1997a). Significantly higher levels of sigmaDDT, sigmaCHL and HCB were found in the NS (1971-1972) stock compared with the StL (1991) animals, and in HCB between the Nfld stock (1971-1972) and the StL (1991) animals, as well as higher p,p'-DDE/sigmaDDT and lower p,p'-DDT/sigmaDDT ratios in the 1991 whales suggesting that temporal trends in these whales between the 1970s and 1990s were consistent with those in other marine mammals in Atlantic Canada. Evidence also suggests that geographical variations in patterns may occur for individuals from two relatively distinct stocks of fin whales in Atlantic Canada (NS, Nfld) and perhaps for one population (StL) related to, but recognisably different from the NS stock. PMID- 11504348 TI - Contamination of polycyclic aromatic hydrocarbons in surface sediments of mangrove swamps. AB - The concentrations of total polycyclic aromatic hydrocarbons (sigmaPAHs) and 15 individual PAH compounds in 20 surface sediments collected from four mangrove swamps in Hong Kong were analysed. sigmaPAH concentrations ranged from 356 to 11,098 ng g(-1) dry weight with mean and median values of 1992 and 1,142 ng g( 1), respectively. These values were significantly higher than those of marine bottom sediments of Hong Kong harbours, suggesting that more PAHs were accumulated in mangrove surface sediments. The concentrations of sigmaPAHs as well as individual PAH compound varied significantly among mangrove swamps. The swamps heavily polluted by livestock and industrial sewage, such as Ho Chung and Mai Po, had much higher concentrations of total PAHs and individual PAH than the other swamps. The PAH profiles were similar among four mangrove swamps, and were dominated by naphthalene (two-ring PAH), fluorene and phenanthrene (three-ring PAH). The mangrove sediments had higher percentages of low-molecular-weight PAHs. These indicated that PAHs in mangrove sediments might originate from oil or sewage contamination (petrogenic input). Ratio values of specific PAH compounds such as phenanthrene/anthracene and fluoranthene/ pyrene, were calculated to evaluate the possible source of PAH contamination in mangrove sediments. These ratios varied among samples, suggesting that mangrove sediments might have a mixed pattern of pyrolytic and petrogenic inputs of PAHs. Sediments collected from Ho Chung mangrove swamp appeared to be more dominated by pyrolytic input while those from Tolo showed strong petrogenic contamination. PMID- 11504349 TI - Organochlorine compounds in common dolphins (Delphinus delphis) from the Atlantic and Mediterranean waters of Spain. AB - Blubber of free-ranging common dolphins (Delphinus delphis) from the northwestern coast of Spain (Atlantic), sampled in 1984 and 1996, and of common dolphins entangled in fishing nets in the southwestern Mediterranean, sampled during 1992 1994, was analysed for organochlorine pollutants. In the Atlantic, concentration of all pollutants was significantly higher in males than in females. The overall tPCB/tDDT ratio in this area was 3.35, which indicates a predominance of industrial inputs over those associated with agriculture. Individuals sampled in 1996 showed significantly lower DDT concentrations but a higher ppDDE/tDDT ratio than those sampled in 1984, which reflects the aging of the environmental load. In the same period, tPCB concentration remained constant and, as a consequence, the tPCB/tDDT ratio more than doubled. In the Mediterranean, the reduced sample size of adult individuals precluded proper statistical investigation of sex related variation. The overall tPCB/tDDT ratio was 1.12, suggesting a higher contribution of pollutants of agricultural origin. Individuals had significantly higher levels of all DDT forms and a higher ppDDE/tDDT ratio than their counterparts from the Atlantic, but similar PCB concentrations. However, the relative frequency of the different congeners in relation to the total PCB load was different in the two areas. This indicates that the two populations do not mix, at least in the short- or medium-term. Organochlorine levels in both areas are at the mid to low end of the range of concentrations detected in other common dolphin populations and in that of other Delphinidae species from the same region. Although the impact of the organochlorine concentrations on the common dolphin populations surveyed cannot be assessed, it is considered unlikely that they have played a significant role in the decline that the species has suffered in recent decades in the western Mediterranean. PMID- 11504350 TI - Lead and zinc in sediments and biota from Maarmorilik, West Greenland: an assessment of the environmental impact of mining wastes on an Arctic fjord system. AB - Lead and zinc levels in sediments and biota from the fjord system surrounding the lead/zinc mine at Maarmorilik, West Greenland, were investigated to evaluate the impact of waste rock and marine-deposited tailings on the marine biota. Concentrations of metal in the sediment were up to 8,922 +/- 622 microg g(-1) (dry wt.) for lead and 19,351+/- 476 microg g(-1) (dry wt.) for zinc. Levels of lead and zinc were also elevated in a suite of monitor organisms. The feeding modes of the organisms were used to explain the sources of metals to the organisms. After closure of the mine, the concentrations of metals in the upper centimetres of the sediments decreased, and a decreased impact of metals on the brown alga Fucus distichus was observed. However, the metals in the sediments still affect the marine biota in the area. PMID- 11504351 TI - Trace metal accumulation and fish pathologies in areas affected by mining and metallurgical enterprises in the Kola Region, Russia. AB - Throughout the Kola region of Russia there has been a substantial increase of metal concentrations in water, which are related to local discharges from metallurgical and mining industry, transboundary transmissions as well as indirect leaching of elements by acid precipitation. This study presents data on the levels of Ni, Cu, Sr, Al, Zn, Co, Mn, Pb, Cd, Hg in the organs and tissues of fish, and evaluates relationships with water chemistry. Special attention is paid to fish pathologies, whose aetiology is related to the accumulation of metals and the associated changes of the elementary ratios within the organism. Ecotoxicological assessment of the copper nickel, strontium and acidification regimes also is considered in this article. In general we observed a large number of lakes that are heavily contaminated by Ni and Cu. Fish in these lakes contain high concentrations of Ni and Cu and display frequent pathologies, mostly associated with the kidneys. In lakes contaminated with Sr, there also are high Sr levels in fish and pathologies associated with skeletal tissues. Exposure to acidified water appears to increase the transport of metals (including Al, Ni and Cu) into fish and hence the toxic effects. PMID- 11504352 TI - Comparing exposure levels by type of welding operation and evaluating the effectiveness of fume extraction guns. PMID- 11504353 TI - Control of paint overspray in autobody repair shops. National Institute for Occupational Safety and Health. PMID- 11504354 TI - Development of an ACGIH construction industry silica exposure database overview. PMID- 11504355 TI - Old millennium reflections--new millennium challenges. PMID- 11504356 TI - Influence of sebum and stratum corneum hydration on pesticide/herbicide collection efficiencies of the human hand. AB - The adhesion of house dust particles and chemical residues to the hands after contact with a variety of surfaces can yield significant exposures to children. Chemicals present on the surface or absorbed into house dust particles may adhere to the hands and subsequently are available for ingestion or absorption through the skin under normal physiological conditions. Contact of a hand with a surface occurs primarily through a thin layer of sebum and sweat on the skin surface. The study presented here examined the effect of the composition of the sebum and sweat layer on the adhesion of a mixture of pesticides and a herbicide to a hand after contact with a house-dust-laden surface. Collection efficiencies on the hand for atrazine and malathion were correlated with sebum levels on the palm of an individual hand. The increase in sebum values measured in this study caused a 13 percent and 8 percent reduction in collection efficiency, respectively, for atrazine and malathion applied to house-dust-laden surfaces. Diazinon and chlorpyrifos collection efficiencies were correlated with skin hydration levels measured on the palm of the individual's hand. The increased skin hydration measured in this study caused a 7 percent and 5 percent reduction in collection efficiency, respectively, for diazinon and chlorpyrifos. PMID- 11504357 TI - Application of radar to detect pedestrian workers near mining equipment. AB - Between 1990 and 1996, 133 accidents occurred and 23 mine workers were killed when haulage trucks used in surface mines collided with another smaller vehicle, a mine structure, or a pedestrian worker. These accidents were caused by a lack of visibility from the cab of the truck. Similar accidents are common with other types of equipment, such as front-end loaders and shovels. There are several methods for improving the operator's awareness of objects or people around the equipment including improved mirror designs, video cameras, and sensor technologies. Researchers at the National Institute for Occupational Safety and Health (NIOSH) are evaluating collision warning systems that are based on radar technology. These systems are mounted on the mining equipment to monitor one or more of the blind areas. An alarm is provided to the operator if an object or person enters the radar's detection area. Tests consisted of mounting the systems on a 50-ton-capacity truck typically used in quarries and a 240-ton-capacity truck used at a surface mine. This article summarizes the test procedure and results of evaluations of several off-the-shelf and prototype radar systems. False alarm rates and reliable detection zones for pedestrians were recorded for various mounting configurations on the rear of the trucks. Mounting radar systems on large equipment presents several challenges; however, the technology does show promise for this application. PMID- 11504358 TI - Surface sampling for a pesticide in dust and small spills of a solid dye. AB - The aim was to determine whether a published sampling technique for loose soil on hard surfaces achieved acceptable efficiency at surface dust/soil coverages of 10 20 mg/ 100 cm2. The sampler was a cordless personal sampling pump operated at 4.0 L/min connected to a cassette containing a filter, the cassette being also connected to a Tygon sampling probe that was moved manually on the surface to be sampled. Rhodamine 6G dye dust was evaluated at 10 mg/100 cm2 coverage at flow rates of 1.0-4.0 L/min. Two National Institute of Standards and Technology (NIST) standard reference materials (SRMs) were used for pesticide experiments: SRM 2711 Montana Soil, and SRM 1649a Urban Dust. The efficiencies for these SRMs were determined uncoated, and coated with 1300 microg chlorpyrifos/g as a Lorsban 4E emulsifiable concentrate formulation. The 3-pass technique sampled both the pesticide-coated and uncoated dust and soil at >79 percent efficiency and at better than 16 percent coefficient of variation (CV) above 15 mg collected mass. Sampling at 20 mg/100 cm2 coverage lowered the CV to < or = 7.1 percent. The dye was sampled with similar efficiency > or = 1.5 L/min, but the CV for the 3-pass technique was <10 percent at 1.5-2.0 L/min, and <20 percent at 1.5-4.0 L/min. The efficiency for a 5-pass technique for the dye exceeded 90 percent at > 1.5 L/min with CVs < 10 percent at 1.5, 2.0, 2.5, and 3.5 L/min. The weight change of the sampling probe and the cassette must be measured for accurate determination of low surface coverages rather than just the weight change of the filter alone. The method allows use of personal sampling equipment for aerosols to measure low coverages of loose dust, soil, and chemical solids on hard surfaces. PMID- 11504359 TI - Effects of peak levels and number of impulses to hearing among forge hammering workers. AB - The purpose of the study was to (1) compare measured and estimated hearing loss among forge hammering workers by applying models for risk assessment based on measurements of impulse noise, and (2) model the hazardous effects of impulse noise on hearing. Noise exposure and hearing loss among forge hammering workers were studied at two forge workshops of an automobile company, where the equivalent sound pressure levels (104 and 105 dB) were the same, but the peak levels and degree of impulsiveness were significantly different. The hearing threshold levels of selected groups of workers (97 and 235 workers) were determined. Comparison between the measured and expected hearing losses defined according to the ISO standard revealed 2 dB difference in excessive hearing loss (1 dB and 3 dB for the workers of workshop 1 and 2, respectively). The excessive hearing loss equals an increase of 3.5 years of exposure. The hearing loss of workers exposed to low impulsive noise could be predicted well using ISO 1999 1990. The hearing loss of workers exposed to high impulsive noise correlated significantly with the peak levels and the number of impulses in combination. PMID- 11504360 TI - The creation of ergonomic data sheets for hazard communication of work-related musculoskeletal disorders. AB - Information on human health hazards caused by chemical exposure has traditionally been supplied through Material Safety Data Sheets (MSDS). This report describes development of an analogous Ergonomic Data Sheet (EDS) to address hazards of work related musculo-skeletal disorders (WRMSD) from use of hand tools. The EDS includes sections on risk factors for WRMSD, design aspects of the tool that decrease risks, precautionary measures, and objective testing of the tool compared to other comparable tools. The EDS can be used within an Ergonomic Communication Program similar to a Hazard Communication Program. PMID- 11504361 TI - A laboratory evaluation of the accuracy and precision of the photovac snapshot portable gas chromatograph and the Drager Chip Measurement System monitor for benzene in air measurements. AB - A laboratory evaluation of the accuracy and precision of two field instruments, the Photovac Snapshot Gas Chromatograph and the Drager Chip Measurement System (CMS) Monitor, both capable of specific determination of benzene, was carried out. The evaluation was based on the generation of a test atmosphere of known concentration of benzene in a stainless steel calibration chamber and simultaneous sampling of the test atmosphere by each instrument. At the same time, the chamber atmosphere was continuously monitored by one or two data logging photoionization detector (PID) hydrocarbon analysers. A series of six, 10 minute charcoal tube air samples of the test atmosphere was also collected over the 1-hour run and analyzed by gas chromatography. Nine different concentrations ranging from 0.25 ppm to 8 ppm were used. Accuracy was evaluated using National Institute for Occupational Safety and Health (NIOSH) criteria. The Photovac GC consistently met the NIOSH recommended accuracy criteria of +/- 25 percent at or below 1 ppm of benzene, whereas the Drager CMS results generally fell slightly outside this criteria. Compared against less stringent accuracy criteria of +/- 35 percent, all Photovac GC results were acceptable but about 40 percent of Drager CMS results were not. The precision of Photovac GC (CV +/- 10%) is better than Drager CMS (CV = +/- 20% to 40%). Both instruments are, however, good field instruments provided their limitations are taken into account in their use. PMID- 11504362 TI - Carbon monoxide, cigarettes and family doctors. AB - AIMS AND BACKGROUND: General practitioners could play a key role in preventive programs against tobacco-related diseases. However, they seldom take action in the office even with minimal advice counselling. Such behaviour might reflect the lack of academic teaching and the lack of practice with motivational and dependence questionnaires, considered basic tools to help smokers to quit successfully. The study was aimed to investigate the awareness of a sample of Italian family doctors as regards tobacco epidemiology and smoking cessation strategies. METHODS: A total of 428 family doctors were administered a questionnaire with a set of questions on their personal smoking habits and on personal initiatives in the office towards smokers. Another set of questions regarded their knowledge on tobacco issues, with special attention to carbon monoxide, which is widely perceived as a very dangerous poison and works as a motivational tool on smokers and adolescents. Carbon monoxide measurement was carried out on all participants to obtain objective data on smoking and to show the feasibility of the test. RESULTS: The percentage of self-reported current smokers among general practitioners was 24%, with a high prevalence of ex-smokers (46%), and 29% of never smokers. Family doctors were more keen to counsel adolescents than adults about tobacco, and they were very interested in continuing medical education on the issue. The doctors who took part in our study showed a surprising limited knowledge of all the issues associated with smoking cessation and prevention such as epidemiology, cigarette characteristics, success rate of smoking cessation programs, Fagerstrom's tolerance questionnaire, safety of nicotine replacement therapy and the knowledge of carbon monoxide as a product of cigarette smoke. CONCLUSIONS: The scenario depicted by our survey underscores the necessity to improve the knowledge and performance of primary care physicians on tobacco-related issues in order to implement primary and secondary prevention in clinical practice. PMID- 11504364 TI - Evaluation of the prognostic accuracy of Astler-Coller's and Jass' classifications of colorectal cancer. AB - AIM: The study aim was to determine whether the Jass classification is superior to that of Astler-Coller (modification of Dukes' stage) in determining prognosis of patients treated for colorectal carcinoma. STUDY DESIGN: The authors used Jass' classification to restage 263 patients who had undergone radical colorectal surgery and classified according to Astler-Coller. RESULTS: The results revealed that: 1) Astler-Coller's classification enables more accurate selection of patient groups where life expectancy can be predicted; 2) Jass' classification enables statistically significant (P <0.05) improved prognostic discrimination of Astler-Coller's B2 patients, for which the probability of nonrecurrence of the disease is around 60%. CONCLUSIONS: Although Astler-Coller's classification is still valid, Jass' classification is useful for the prognostic discrimination of Astler-Coller's B2 patients. PMID- 11504363 TI - Cytoreductive surgery followed by intraperitoneal hyperthermic perfusion in the treatment of recurrent epithelial ovarian cancer: a phase II clinical study. AB - AIMS AND BACKGROUND: The optimal salvage therapy for recurrent ovarian carcinoma has not been clearly established. Response to second-line chemotherapy is low, with a short median survival (8.8-15 months). We investigated the effect of an aggressive approach consisting of surgery followed by intraperitoneal drug delivery and local hyperthermia. PATIENTS AND METHODS: In a phase II clinical study, 27 patients with advanced/recurrent ovarian carcinoma were treated with cytoreductive surgery and intraperitoneal hyperthermic perfusion. Median patient age was 53 years (range, 30-67) and mean follow-up was 17.4 months (range, 0.3 36.0). Patients had been surgically staged and heavily pretreated with cisplatin based, taxol-based or taxol/platinum-containing regimens. Nineteen (70%) patients were cytoreduced to minimal residual disease <2.5 mm. The intraperitoneal hyperthermic perfusion was performed with the closed abdomen technique, using a preheated polysaline perfusate containing cisplatin (25 mg/m2/L) + mitomycin C (3.3 mg/m2/L) through a heart-lung pump (mean flow of 700 mL/min) for 60 min in the hyperthermic phase (42.5 degrees C). RESULTS: Two-year overall survival was 55%. Median times to overall progression and local progression were 16 months and 21.8 months, respectively. Variables that affected the overall survival or time to progression were as follows: residual disease (P = 0.00025), patient age (P = 0.04), and lag time between diagnosis and cytoreductive surgery + intraperitoneal hyperthermic perfusion (P = 0.04). Treatment-related morbidity, mortality and acute toxicity (grade II-III) rates were 11%, 4% and 11%, respectively. Eight (89%) of 9 patients had ascites resolution. CONCLUSION: Our results suggest that cytoreductive surgery + intraperitoneal hyperthermic perfusion is a well tolerated, feasible and promising alternative in the management of selected patients with recurrent ovarian cancer, but further randomized controlled studies are needed in order to confirm our findings. PMID- 11504365 TI - Roach's mathematical equations in predicting pathological stage in men with clinically localized prostate cancer. AB - AIMS AND BACKGROUND: The therapeutic choice in patients with clinically localized prostate cancer depends on preoperative clinical stage. Diagnostic instruments currently available for such an evaluation--considered separately--have not shown enough efficacy. Roach has recently introduced three simple mathematical equations that--on the basis of prostate-specific antigen and the biopsy Gleason score--are aimed at calculating the definitive pathological stage. We retrospectively analyzed our radical prostatectomy data base to assess the accuracy of the equations in predicting the final stage in patients with clinically localized prostate cancer. METHODS STUDY DESIGN: The study included 173 patients who had undergone radical retropubic prostatectomy at our Institution. Patients were divided into 25 groups, depending on preoperative PSA and the biopsy Gleason score. The risk of extracapsular neoplastic growth, seminal vesicle involvement and lymph node involvement was calculated for each group by means of Roach's equations. On the basis of definitive histological examinations, we compared the expected risk to the observed risk. RESULTS: The observed risk fell within the interval of expected risk in 16 of 17 groups (94%) regarding the evaluation of extracapsular growth, in 15 of 17 (88%) regarding the analysis of seminal vesicle involvement, and in 14 of 17 (82%) regarding the evaluation of lymph node involvement. Therefore, the observed event was in agreement with the expected event in 45 of 51 groups (88%). CONCLUSIONS: The equations represent a practical and effective instrument for preoperative clinical staging in patients with localized prostate cancer. By means of these mathematical formulas, one can assess the correct prognosis and--above all--plan the best therapeutic approach. PMID- 11504366 TI - Salvage treatment with epirubicin and/or paclitaxel in metastatic breast cancer patients relapsed after high-dose chemotherapy with peripheral blood progenitor cells. AB - AIMS AND BACKGROUND: To evaluate feasibility and efficacy of paclitaxel as a single agent or in combination with epirubicin in breast cancer taxane-naive patients who have failed previous high-dose chemotherapy. METHODS: Since February 1995, we have treated 32 patients in first relapse or progression after high-dose chemotherapy. Nineteen patients had metastatic breast cancer, 12 more than 3 involved axillary lymph nodes, and 1 inflammatory breast cancer at inclusion to the program. The median time to relapse after high-dose chemotherapy was 12 months (range, 2-43). At relapse, 12 patients were treated with epirubicin (90 mg/m2) plus paclitaxel (175 mg/m2) administered on day 1 every 21 days. In 20 patients who had previously received more than 350 mg/m2 of a cumulative dose of epirubicin and in one patient pretreated with chemotherapy containing mitoxantrone, we employed paclitaxel (175 mg/m2) alone. A median number of five courses was administered (range, 2-10). RESULTS: The overall response rate after 3 courses (29 of 32 patients were assessable) was 55% and after 6 courses (21 of 32 patients were assessable) was 57%. The median time to progression was 7 months (95% CI, 5.7-9.2), and median survival was 27.5 months (95% CI, 17.8-37.0). Toxicity was recorded for 180 cycles (epirubicin + paclitaxel for 62 cycles and paclitaxel alone for 118 cycles). The main toxicity in both regimens was hematologic. We observed WHO grade 3-4 neutropenia (in 8 patients, 25%), for which G-CSF (5 microg/kg/day s.c.) was employed. WHO grade 3-4 thrombocytopenia occurred in 2 patients (6%) and WHO grade 3 anemia in 1 patient (3%). CONCLUSIONS: Our study showed that paclitaxel (alone or in combination with epirubicin) is feasible as salvage treatment in heavily pretreated patients. PMID- 11504367 TI - Results of a prospective study with high-dose etoposide, thiotepa and carboplatin and peripheral blood stem cell rescue for high-risk stage II-IIIA and selected stage IV breast cancer patients. AB - AIMS AND BACKGROUND: To investigate the safety and efficacy of a high-dose chemotherapy regimen with etoposide, carboplatin and thiotepa in high-risk stage II-IIIA breast cancer and in responsive metastatic patients. STUDY DESIGN: From April 1992 to December 1998, 24 patients with high-risk stage II-IIIA breast cancer (> or = 9 positive nodes) and 9 responsive metastatic patients were enrolled in the trial. After induction chemotherapy with an anthracycline-based regimen, peripheral blood stem cells were mobilized with cyclophosphamide (7 g/m2) and G-CSF (5-16 microg/kg/s.c./day). The high-dose chemotherapy regimen consisted of etoposide (1000 mg/m2), carboplatin (800 mg/m2) and thiotepa (500 mg/m2). At the end of the high-dose chemotherapy, all stage II-IIIA patients received radiotherapy to the breast or chest wall and draining nodes; stage IV patients were irradiated to sites of disease, if feasible. All ER+ and/or PgR+ patients were treated with hormone therapy. RESULTS: For stage II-IIIA high-risk patients, the median follow-up was 4.36 years (range, 1.93-6.94), and the Kaplan Meier estimate at 5 years of disease-free survival and overall survival was 54.8 +/- 11% SE and 76.73 +/- 9.4% SE, respectively. For metastatic patients, the median follow-up was 4.93 years (range, 4.15-7.95), and the Kaplan-Meier estimate at 5 years of progression-free survival and overall survival was 22.2 +/- 13.9% SE and 76.2 +/- 14.8% SE, respectively. No treatment-related deaths were observed. CONCLUSIONS: Our results are comparable to those obtained in other high dose chemotherapy trials but do not seem to be superior to conventional-dose therapy given to similar patients. PMID- 11504368 TI - Radioimmunoguided breast surgery using radiolabeled antibody NR-LU-10 FAB: a pilot study. AB - AIM AND BACKGROUND: Radioimmunoguided surgery using radiolabeled NR-LU-10 Fab was evaluated as a method of intraoperative breast cancer detection. METHODS: Breast cancer patients were injected intravenously with 125I (74 MBq) labeled NR-LU-10 Fab (5 mg) and then underwent tumor excision 2, 4 or 7 days later, during which time the gamma detector probe was used to evaluate the primary tumor for evidence of radioactive uptake. RESULTS: Intraoperative probing revealed tumor localization in 7 of 10 patients (70%). Gamma probe counts of the excised tumor were elevated in all patients, although high counts in surrounding non-malignant tissue obscured the ability to detect the tumor in vivo in 3 patients. One patient with bilateral breast cancer was found to have a separate focus of occult tumor in each breast using the gamma detector probe. CONCLUSIONS: Radiolabeled NR LU-10 Fab possesses favorable pharmokinetics and tumor-binding ability as a targeting agent. However, binding to non-malignant tissue limits its role in the intraoperative evaluation of tumor margins in breast cancer patients. Its role in other malignancies should be explored. PMID- 11504369 TI - Radiotherapy-related fatigue: how to assess and how to treat the symptom. A commentary. AB - Acute and chronic radiotherapy-related fatigue occurs in up to 80% and 30%, respectively, of patients undergoing irradiation for cancer. Frequently, the symptom is not expected by the patients and is underestimated by medical and nursing staff. Fatigue can affect global quality of life more than pain, sexual dysfunction and other cancer- or treatment-related symptoms. Its etiology and correlates are not clear. Published reports are mainly descriptive, and in many of them numerous methodological biases are present. One of the limitations is lack of a standard method of assessment that could simplify the comparison between different series. In the last decade, modern instruments have been designed to measure fatigue. They include uni- and multidimensional tools. Use of these specific instruments is highly recommended for research on radiation related fatigue. In daily practice when time is limited, simple assessment is necessary. For example, systemic use of plain and easily understandable questions about fatigue, its level and impact on daily life could be sufficient and reliable. Therapeutic strategies for radiotherapy-induced fatigue have not yet been clearly defined, but a few randomized studies have been recently published. Physical exercise, group psychotherapy and relaxation therapy have been demonstrated to be effective. Moreover, pharmacological treatment of concomitant disturbances (anemia, pain, insomnia, depression, dehydration, infection, malnutrition) and other radiotherapy side effects (diarrhea, hormonal insufficiency etc.) should be considered. Further methodologically correct studies are warranted to better define the causes, optimal prevention, assessment and management of this symptom. PMID- 11504371 TI - An experience of stereotactic radiation therapy for primary intracranial choriocarcinoma. AB - We report on a patient with choriocarcinoma in the pineal region who was successfully treated with stereotactic radiation therapy (SRT). The increased level of serum human chorionic gonadotropin (HCG) was lowered during chemotherapy with etoposide, cisplatin, and ifosfamide. However, HCG was not normalized and magnetic resonance images still showed an enhanced tumor mass with gadolinium. The patient underwent SRT of 40 Gy at an 80% isodose line per 10 fractions over two weeks, followed by conventional craniospinal irradiation of 32.4 Gy. The level of HCG dropped below the detection limit. The patient has been in good condition for more than four years after the completion of treatment, without any signs of recurrence. We propose SRT as a valid treatment option for malignant germ cell tumors in the pineal region. PMID- 11504370 TI - On the delineation of the gross tumor volume and clinical target volume for head and neck squamous cell carcinomas. AB - Gross tumor volume (GTV) and clinical target volume (CTV) delineation on planning computed tomography (pICT) for head and neck squamous cell carcinomas can be troublesome. We highlight the factors which can be crucial for the radiation oncologist in delineating GTV and CTV on pICT and provide some pratical solutions. Regarding GTV, uncertainties are correlated with transfer of information collected by physical examination and diagnostic radiology to pICT. Moreover, reproducibility of delineation can also be highly variable, particularly when diagnostic imaging quality and pICT quality are poor. Once the prescription has been made, clinical target volume identification on pICT is rarely straightforward. Whereas there are some data about the location of major lymph node stations of the neck, there are no reported guidelines on how to draw subclinical extention of primary head and neck tumors on pICT. Such volumes can be derived from those currently included in simulator films or from those addressed by the surgeon. Some examples are provided. A particular situation is represented by the adjuvant setting, when the primary tumor is removed (by surgery) or reduced (by chemotherapy). In conclusion, this paper shows some major problems associated with identification of GTV and CTV on pICT. Apart from selected cases, the use of pICT for target volume delineation (and thus for field shaping) for head and neck squamous cell carcinoma is still to be considered investigational. PMID- 11504372 TI - Ret oncogene protein expression in papillary thyroid carcinoma and related lesions. AB - BACKGROUND: Activation of Ret oncogenes, particularly Ret/PTC, has been identified in papillary thyroid carcinoma (PTC). The purpose of this study was to investigate the immunostaining pattern of Ret oncogene protein in PTC and nodular non-PTC lesions with a fine chromatin pattern. MATERIALS AND METHODS: Ninety three PTC and 139 nodular non-PTC lesions were microscopically reviewed to identify the nuclear changes of "limited nuclear features of PTC" (focal nuclear grooves, nuclear inclusions or optically clear nuclei) and areas of infiltrating carcinoma (IC) and were submitted for immunostaining with Ret oncogene protein antiserum. RESULTS: Immunoreactivity for Ret protein ranged from negative in follicular adenoma (FA) with a coarse chromatin pattern, to negative or weak reactivity in FA with a fine chromatin pattern, to strong reactivity in PTC with areas of infiltrating carcinoma (IC). In FA with fine chromatin, FA and follicular carcinoma (FC) containing an admixture of areas of coarse and fine chromatin, areas with nuclear changes with "limited nuclear features of PTC" displayed varying degrees of immunoreactivity. The intensity of immunostaining varied with the degree of nuclear change. The noninvasive component of PTC with IC usually showed more extensive and stronger reactivity than PTC without IC. PTCs with and without IC were associated with a rate of lymph node metastasis of 48% and 3%, respectively. CONCLUSIONS: The expression of Ret oncogenes (Ret/PTC, other unknown variants or wild type) is focally or extensively present in all PTC with IC. The degree of immunoreactivity is likely to be proportional to the potential for lymph node metastasis of PTC. In the context of this study and due to the specificity of Ret oncogenes, it is likely that nodular non-PTC lesions with a fine chromatin pattern and focal positive reactivity for Ret oncogene represent PTC-related lesions. PMID- 11504373 TI - Cadherins and integrins in renal cell carcinoma: an immunohistochemical study. AB - AIMS AND BACKGROUND: The aim of this study was to determine the expression of cadherins and integrins in renal cell carcinoma (RCC) and their relationship with tumor morphology and TNM status. METHODS: Cadherin and integrin expression was investigated using an indirect immunoperoxidase technique, applying antibodies to E-, N-, P- and VE-cadherin and to alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, and alpha(v) integrin subunits. Correlation of semiquantitatively scored adhesion molecule levels with histopathological parameters (cytology, growth pattern, nuclear grade) and TNM status was performed for 24 RCCs (17 clear cell, 3 granular, 3 spindle cell and 1 chromophobe cell type according to the WHO classification). RESULTS: E-cadherin and N-cadherin were present in most cases (88% and 67%, respectively) and were usually coexpressed. T3 RCCs displayed higher E-cadherin and N-cadherin levels than T1/T2 tumors regardless of tumor grade, suggesting that impairment of their function might exist without actual loss from tumor cells. P-cadherin was found focally in two RCCs only, while VE cadherin was present on stromal vessel endothelium in five tumors, showing no differences with regard to cell type, growth pattern, tumor grade or TNM status. All integrins were present in the studied RCCs (ranging from 12% for alpha5 to 79% for alpha3), including those that are normally absent from adult kidney tissue (alpha4 and alpha5). Tumors of higher grade showed increased alpha(v) and decreased alpha6 levels, while RCCs with metastases less often showed diffuse alpha3 presence and never expressed alpha5 integrin. CONCLUSIONS: Our results suggest that the level of expression of N-cadherin and some integrins (most notably alpha3, alpha6 and alpha5) is associated with the capacity of RCC for local and distant spread, regardless of tumor grade. PMID- 11504374 TI - Immunohistochemical study of 49 cutaneous melanomas: p53, PCNA, Bcl-2 expression and multidrug resistance. AB - AIMS AND BACKGROUND: Thickness and level of invasion are the main morphological elements for an approximate but not sufficiently sensitive prognostic evaluation of cutaneous melanomas. By using immunohistochemical methods it is possible to detect biological markers related to prognosis. We have studied p53, PCNA, Bcl-2 and P-gp expression in 49 primary cutaneous melanomas. MATERIALS: We used the immunophosphatase APAAP immunohistochemical method. The percentage of labeled cells (according to four classes of positivity: <5%; 5-25%; 25-50%; >50%) and the localization of immunoreactivity were expressed for each marker. Statistical analysis was performed to determine the correlations between markers and level or thickness of melanomas. RESULTS: We found a good correlation between p53 expression and melanoma thickness (P <0.005), PCNA and P-gp expression. No relationship was observed between Bcl-2 expression and the different variables considered or other markers. CONCLUSIONS: Our data seem to indicate an unfavorable prognostic role of higher nuclear p53 expression. However, we believe that our results need to be integrated with patients' clinical follow-up and with the study of the expression of these markers in benign melanocytic lesions to gain more accurate information about their prognostic significance. PMID- 11504375 TI - Deep venous thrombosis of the extremity diagnosed by color Doppler ultrasonography after isolated limb perfusion. AB - AIM: Isolated limb perfusion (ILP) is used to treat locally advanced sarcoma and melanoma of the extremities. ILP is associated with a 1.7% to 10% incidence of deep venous thrombosis (DVT). The aim of the study was to examine the effect of ILP on the peripheral venous system and to evaluate the diagnostic accuracy of color Doppler ultrasonography (US). METHODS: A total of 26 patients with locally advanced sarcoma or melanoma of the extremities received a total of 38 ILP. The patients were evaluated preoperatively and postoperatively by color Doppler US as a noninvasive venous measurement. We used a color Doppler US system (SSA-270A, Toshiba) with a 7.5-MHz linear transducer for gray-scale imaging and a 5-MHz vascular transducer for color Doppler imaging. RESULTS: Seventeen patients underwent single ILP while the others were treated with multiple ILPs. Color Doppler US showed a reflux flow in three (7.9%) patients and DVT occurred subsequently in these patients. CONCLUSIONS: Color Doppler US is a noninvasive and clinically useful diagnostic technique in the diagnosis of extremity DVT. We recommend anticoagulant therapy in patients with reflux on ultrasonographic examination. PMID- 11504376 TI - Two cases of histopathologically advanced (stage IV) early gastric cancers. AB - We report two cases of early gastric cancer with distant metastases (stage IV). At our institute 1428 cases of primary gastric cancer were resected between 1980 and 1997; 536 were diagnosed as early gastric cancer based on the resected specimens (304 cases of mucosal cancer, Tis--TNM classification--and 232 of submucosal cancer, T1). 528 of these 536 cases were classified as histological stage I, six as stage II, none as stage III and two as stage IV. The incidence of stage IV early gastric cancer was 0.14% of all gastric cancers and 0.37% of the early gastric cancers. The two patients with stage IV early gastric cancer were women. Both tumors were defined as early cancer because they were confined to the submucosa. One was a type 0 IIc + III early cancer, histologically classifiable as a small, moderately differentiated adenocarcinoma (tub2 according to the Japanese Classification of Gastric Carcinoma, G2; TNM classification: ICD-O C16), size 10 x 8 mm; the other was a surface spreading type 0 IIc, classifiable as a signet-ring cell carcinoma (sig, G3), size 50 x 35 mm. Stage IV factors were N3 in the first and ovarian metastasis (Krukenberg tumor) in the second case. PMID- 11504377 TI - Epithelioid angiomyolipoma: a case report. AB - Epithelioid angiomyolipoma is a recently recognized tumor entity. We report a case of epithelioid angiomyolipoma of the kidney that occurred in a 70-year-old female. The patient presented with massive abdominal hemorrhage and shock. There was no history of tuberous sclerosis complex. The tumor consisted of smooth muscle, adipocytes, small-to-medium-sized hyalinized blood vessels and numerous pleomorphic epithelioid cells with clear to eosinophilic cytoplasm. Mitosis was exceptional and no glandular pattern was recognizable. Immunohistochemically the epithelioid cells were strongly positive for HMB45 and CD68, focally positive for S-100, actin and vimentin, and negative for epithelial markers including epithelial membrane antigen and cytokeratins. Previous reports in the English literature suggest that epithelioid angiomyolipoma may have malignant potential. In our case, at six months from surgery the patient is alive and well without evidence of relapse or metastasis. PMID- 11504378 TI - Acute cardiotoxicity during capecitabine treatment: a case report. AB - Capecitabine (Xeloda, Roche, Monza), a fluoropyrimidine carbamate, is an orally administered drug that delivers fluorouracil (5-FU) selectively to the tumor. The drug has demonstrated activity in metastatic breast cancer, pancreatic cancer and colorectal cancer. In this case report the authors describe an unusually and reversible cardiac side effect which occurs to 39-year-old patient treated with capecitabine 2000 mg/m2/day for advanced gastric cancer. It is important to note that the safety data from clinical trials indicate that capecitabine has a toxicity profile typical of infused fluoropyrimidines. However, none of the studies described cardiac side effects. PMID- 11504379 TI - Parachordoma: a case report. AB - Parachordoma is a very rare soft tissue tumor with histological features similar to chordoma and chondrosarcoma. It should be distinguished from metastatic chordoma and extraskeletal myxoid chondrosarcoma because of its different treatment and prognosis. In this paper we report one case of parachordoma in a 20 year-old female patient. The tumor occurred in the subcutaneous tissue of the left hand as a painless, fixed, slow-growing mass. Pathologic analysis revealed a tumor composed of lobules of cells with variably vacuolated cytoplasm (physaliphorous cells) separated by fibrous septa, predominantly arranged in peculiar small or large alveolar structures. Immunohistochemistry showed positive staining of the tumor cells with cytokeratin 8/18, S-100 protein and vimentin. The patient is well and without recurrence 20 months after surgery. PMID- 11504380 TI - Differential effects of retinoids on proliferation of bovine mammary epithelial cells in collagen gel culture. AB - The effects of increasing concentrations of retinol, retinal and retinoic acid on proliferation of bovine mammary epithelial cells were investigated in collagen gel cultures. All retinoids significantly inhibited proliferation of mammary epithelial cells. The relative inhibitory potency of the retinoids was: retinoic acid > retinal > retinol. Maximal inhibition at 10 microg/ml corresponded to a 75 95% inhibition of proliferation obtained in basal medium. Retinol, retinal and retinoic acid also inhibited proliferation of cells growth-stimulated with insulin-like growth factor-I (IGF-I). Retinoids in highest concentrations (10 microg/ml) inhibited 68-85% of proliferation of cells obtained in culture medium containing 25 ng IGF-I/ml. Retinol and retinoic acid also inhibited proliferation of cells growth-stimulated by insulin and other growth factors from the IGF growth factor family (des(1-3)IGF-I and IGF-II), as well as growth factors from the epidermal growth factor family (EGF and TGF-alpha), with retinoic acid being more effective than retinol. At a concentration of 100 ng/ml retinol and retinoic acid inhibited respectively 24-38 and 44-52% of mammary cell proliferation stimulated by growth factors of the IGF family, and at 10,000 ng/ml, 61-71% of cell proliferation was inhibited. The growth-stimulating effect of insulin, EGF and TGF-alpha was inhibited 42-64% by retinol and retinoic acid at 100 ng/ml, and 64-84% at 10,000 ng/ml. The present results show that retinol, retinal and retinoic acid are potent inhibitors of bovine mammary epithelial cell proliferation. It is suggested that retinoids may have concentration-dependent roles in regulation of pubertal mammary growth and development, indicating that the milk yield potential of heifers may be affected by vitamin A status. PMID- 11504381 TI - Lactation failure in crossbred Sahiwal Friesian cattle. AB - Milk producers in Malaysia make extensive use of crossbred Sahiwal Friesian dairy cattle. These animals have, however, been found susceptible to lactation failure. A survey of cows in an experimental herd of F1 Sahiwal Friesian animals indicated that, in 30% of animals, milk yield decreased to negligible levels within the first 8 weeks post partum. Lactation failure was associated with a progressive increase in the amount of residual milk left in the udder after normal milking. By week 3 of lactation, residual milk volume was significantly greater than that in animals that, based on previous lactation history were not susceptible to lactation failure, and accounted for up to 30% of milk available at the morning milking. The cellular consequences of residual milk accumulation were evident in the activities of acetyl-CoA carboxylase, fatty acid synthetase and galactosyltransferase, key enzyme markers of cellular differentiation, which decreased in glands undergoing lactation failure and were lower than values measured in tissue of control cows. Mammary cell number, estimated by tissue DNA content, was also reduced in animals undergoing lactation failure. These indices of mammary development indicate that lactation failure is the result of premature involution in susceptible animals. Premature involution is a predictable consequence of progressive milk stasis in failing lactation, and attributable to an increase in autocrine feedback by inhibitory milk constituents. The progressive increase in residual milk is, on the other hand, unlikely to be attributable to impaired mammary development. Measurements of milk storage during milk accumulation showed no differences between control and lactation failure cows in the distribution of milk between alveolar and cisternal storage compartments. We conclude that lactation failure in Sahiwal Friesian cows is due to a failure of milk removal, and probably the result of an impaired milk ejection reflex rather than to the glands' milk storage characteristics. PMID- 11504382 TI - Effect of suckling on the release of oxytocin, prolactin, cortisol, gastrin, cholecystokinin, somatostatin and insulin in dairy cows and their calves. AB - The aim of the present study was to examine how different types of early interaction between dairy cows and their calves influence milking/suckling related hormone release in the cows and sucking/bucket-drinking related hormone release in calves. Eighteen cows of the Swedish Red and White breed were studied during the first week after parturition. The cows were machine milked twice daily, and allotted to one of three treatments: [1] cow and calf were kept together and the cow was allowed to suckle the calf for 30 min about 1 h before each milking; [2] cow and calf were kept together and the calf was bucket fed twice daily; and [3] cow and calf were separated immediately after parturition, and the calf was kept in a single box and was bucket fed twice daily. Blood samples were collected around day 7 from both cows and calves. The plasma levels of oxytocin, prolactin, cortisol, gastrin, cholecystokinin (CCK), somatostatin and insulin were analysed. In the cows the levels of oxytocin, prolactin and cortisol were influenced by all three treatments, except for the level of cortisol which did not respond to suckling. The main finding was that the release of oxytocin was as significantly greater during suckling compared with machine milking. In the calves, the hormone levels were also influenced by the different milk feeding routines. The plasma concentrations of oxytocin, gastrin, CCK and insulin increased in response to milk ingestion in all treatments. However, during sucking, the increase of oxytocin was significantly greater than during bucket drinking. In addition, a strong correlation between oxytocin and insulin was found in response to sucking. Further, significant increases in prolactin and somatostatin, and a decrease in cortisol were found during sucking. The level of somatostatin also increased in response to bucket feeding when calves were kept separately. During bucket feeding, no significant correlation was found with oxytocin, but strong correlations between the gastrointestinal hormones gastrin, CCK, somatostatin and insulin were seen. Together these data suggest that different hormonal patterns were triggered in the cows by suckling and milking and in the calves by sucking and bucket drinking. This is further supported by different correlation patterns observed in the calves in response to sucking and bucket feeding. The present findings imply that management routines for cows and calves during the first week after parturition have consequences for the physiology of the animals. PMID- 11504383 TI - Contamination of pasteurized milk by Bacillus cereus in the filling machine. AB - The contamination of pasteurized milk by Bacillus cereus during the filling process was studied in two dairy plants. Samples of pasteurized milk were taken at four different sites along the production line. The samples were stored at 7 degrees C for 7 d, or at 10 degrees C for 5 d, before plate counting and random selection of B. cereus isolates. Isolates of B. cereus were typed by the polymerase chain reaction (PCR)-based method randomly amplified polymorphic DNA (RAPD). Samples taken at three different sites between the pasteurizer and the filling machine were all holding similar low concentrations of B. cereus, while an increase of the B. cereus count was seen in the consumer packages. More B. cereus of different RAPD types was growing in the consumer packages than in samples taken just before the filling machine. Several RAPD types found in the consumer packages were not detected in the samples taken just before the filling machine. PMID- 11504384 TI - Heterogeneity of caprine kappa-casein macropeptide. AB - The heterogeneity of caprine caseinmacropeptide (CMP) was determined by means of treatments with neuraminidase and acid phosphatase and analyses by anion exchange FPLC and reversed-phase (RP)-HPLC, with on-line and off-line electrospray ionization mass spectrometry. The main CMP components were two non-glycosylated and di-phosphorylated forms, as well as two other mono-phosphorylated species, each corresponding to a genetic variant of caprine kappa-casein due to the silent substitution Ile/Val at position 119. Asialo-aglyco mono- and di-phosphorylated forms were found in the ratios 8-14% and 86-92%, respectively. Approximately 36% of caprine CMP was glycosylated. Based on the obtained molecular masses, the occurrence of tri-, di- and monosaccharide-containing di-phosphorylated CMP are reported, assuming that N-acetylgalactosamine, galactose, N-acetyl and N glycolylneuraminic acids would constitute the main monosaccharides of caprine CMP. CMP microheterogeneity due to the genetic polymorphism was also observed in the glycosylated forms. PMID- 11504385 TI - Genetic polymorphism of the caprine kappa casein gene. AB - Polymorphism in the goat kappa casein gene was studied using the base excision sequence scanning (BESS) method and sequencing. Seven polymorphic sites, corresponding to single nucleotide transversions were detected. Three of these were silent mutations while the other four produced amino acid substitutions. The association between these polymorphic sites was investigated, which resulted in the identification of three goat kappa casein alleles, designated A, B, and C. Protocols for rapid genotyping of the C variant were developed by polymerase chain reaction-restriction fragment length polymorphism using Alw44I and BseNI restriction endonucleases. The occurrence of this allele was found to be very low in Spanish breeds but more frequent in the French Saanen goat. Further studies among different goat populations are necessary to establish the distribution of these alleles and their effects on the quality and functional properties of milk. PMID- 11504386 TI - Goats' milk of defective alpha(s1)-casein genotype decreases intestinal and systemic sensitization to beta-lactoglobulin in guinea pigs. AB - Contradictory results have been reported on the use of goats' milk in cows' milk allergy. In this study the hypothesis was tested, using a guinea pig model of cows' milk allergy, that these discrepancies could be due to the high genetic polymorphism of goats' milk proteins. Forty guinea pigs were fed over a 20 d period with pelleted diets containing one of the following: soyabean proteins (group S), cows' milk proteins (group CM), goats' milk proteins with high (group GM1) or low (group GM2) alpha(s1)-casein content. Parenteral sensitization to GM1 and GM2 proteins as also assessed. The sensitization was measured (1) by systemic IgG1 antibodies directed against bovine or caprine beta-lactoglobulin (beta-lg), alpha-lactalbumin (alpha-la) and whole caseins, and (2) by intestinal anaphylaxis measured in vitro in Ussing chambers, by the rise in short-circuit current (delta Isc) in response to milk proteins. Guinea pigs fed on CM and GM1 developed high titres (> 1500) of anti-beta-lg IgG1, with an important cross reactivity between goat and cow beta-lg. However, in guinea pigs fed on GM2, anti-goat beta-lg IgG1 antibodies were significantly decreased compared with GM1 guinea pigs (mean IgG1 titres were 546 and 2046 respectively), and the intestinal anaphylaxis was significantly decreased (3.5+/-4.5 microA/cm2) compared with that observed in GM1 guinea pigs (8.3+/-7.6 microA/cm2). Animals receiving GM1 or GM2 proteins via the parenteral route developed a marked sensitization. These results suggest that the discrepancies observed in the use of goats milk in cows' milk allergy could be due, at least in part, to the high genetic polymorphism of goats' milk proteins. PMID- 11504387 TI - Detection of cows' milk in goats' cheeses inferred from mitochondrial DNA polymorphism. AB - A new polymerase chain reaction (PCR)-based method was developed to detect cows milk in goat cheese. This method is based on mitochondrial DNA (mtDNA) control region sequence variations. DNA extractions from 150 mg of cheese were carried out using a spin column-based method. Subsequent PCR amplifications of DNA were performed with cow specific primers, demonstrating the ability to detect cows' milk in a variety of cheeses. This simple approach provides high quality DNA, and is shown to be very sensitive, with a detection limit of less than 0.1% of cows' milk. Analysis of an agarose gel digital image allows a rough estimation of the percentage of cows' milk used in adulteration. PMID- 11504388 TI - Effect of added salt and increase in ionic strength on skim milk electroacidification performances. AB - Bipolar-memibrane electroacidification (BMEA) technology which uses the property of bipolar membranes to split water and the demineralization action of cation exchange membranes (CEM), was tested for the production of acid casein. BMEA has numerous advantages in comparison with conventional isoelectric precipitation processes of proteins used in the dairy industry. BMEA uses electricity to generate the desired ionic species to acidify the treated solutions. The process can be precisely controlled, as electro-acidification rate is regulated by the effective current density in the cell. Water dissociation at the bipolar membrane interface is continuous and avoids local excess of acid. In-situ generation of dangerous chemicals (acids and bases) reduces the risks associated with the handling, transportation, use and elimination of these products. The aim of this study was to evaluate the performance of BMEA in different conditions of added ionic strength (p(added) = 0, 0.25, 0.5 and 1.0 M) and added salt (CaCl2, NaCl and KCl). The combination of KCl and p(added) = 0.5 M gave the best results with a 45% decrease in energy consumption. The increased energy efficiency was the result of a decrease in the anode/cathode voltage difference. This was due to an increase of conductivity, produced by addition of salt, necessary to compensate for the lack of sufficiently mobile ions in the skim milk. However, the addition of salts, irrespective of type or ionic strength, increased the required operation time. The protein profile of isolates were similar under all experimental conditions, except at 1.0 M-CaCl2. PMID- 11504389 TI - Correlation of base consumption with the degree of hydrolysis in enzymic protein hydrolysis. AB - It is fairly easy to control the enzymic hydrolysis of proteins in alkaline conditions by measuring the base consumption required to keep the pH constant in the reactor. Unfortunately, however, base consumption is not related in any simple way to the degree of hydrolysis reached at any given moment and to establish this relationship it is essential to find out the mean pK of the alpha amino groups released during the hydrolytic process. We have shown here that the correct mean pK value varies according to the pH of the working conditions and that the relationship between these values may depend upon the kind of protein and protease used. We have put forward a method for determining this relationship experimentally by using a given protein-protease system, consisting of an alkaline titration of the raw protein and when partially hydrolysed. We have tested the results predicted by our theoretical model by applying it to the hydrolysis of whey proteins with a bacterial protease from Bacillus licheniformis at 50 degrees C, pH 8.0. This model can easily be applied to any hydrolytic process involving the appearance of functional groups that are partially protonizable under the working conditions in question in order to follow the kinetics of the reaction via the consumption of the neutralizing agent required to keep pH constant. PMID- 11504390 TI - Thermal inactivation kinetics of bovine cathepsin D. AB - Cathepsin D, the principal indigenous acid proteinase in bovine milk, is a lysosomal proteinase, which exists in milk in four forms, including the inactive zymogen procathepsin D. The thermal inactivation kinetics of bovine cathepsin D, isolated from spleen and milk, were studied under isothermal conditions, using a specific HPLC assay to determine residual activity. Inactivation of the blood enzyme preparation followed first order kinetics, with z-values in phosphate buffer (pH 6.7) and skimmed milk of 6.5 and 7.6 degrees C, respectively, the enzyme being far more stable in the latter environment. Inactivation kinetics of the enzyme purified from milk were more complex, and could be best approximated by a double exponential model. Again, stability was higher in milk than in buffer. The double exponential model may indicate differing heat stabilities of isoforms of the enzyme, or stabilization of the enzyme by some milk constituent. It is clear that the enzyme can survive, at least partially, processes such as heating at 55 degrees C for 30 min during manufacture of high-cook cheese varieties (45% survival), and HTST pasteurization (8% survival), and thus may contribute to proteolysis in a range of dairy products. PMID- 11504391 TI - Molecular self-assembly of partially hydrolysed alpha-lactalbumin resulting in strong gels with a novel microstructure. AB - Gelation of alpha-lactalbumin (alpha-la) incubated with a protease from Bacillus licheniformis (BLP) at 50 degrees C for 4 h was monitored using small oscillatory shear and the large deformation properties of final gels were characterized by uniaxial compression. Transmission electron microscopy was used to visualize the microstructure. Gels made from alpha-la (10 g/l) using BLP were almost transparent, although somewhat whitish, and they were more than 20 times stiffer (measured as complex modulus) than equivalent gels made from beta-lactoglobulin (beta-lg) at the same concentration. The microstructure of the gels consisted of non-branching, apparently hollow strands with a uniform diameter close to 20 nm, similar in overall structure to microtubules. Adding Ca2+ in amounts of 50 or 100 mM changed the spatial distribution of the strands and resulted in a reduction in the failure stress recorded in uniaxial compression. Apart from affecting the microstructure, Ca2+ was shown to be essential for the formation of the gels. It is proposed. that the mechanism behind the self-assembly of the partially hydrolysed alpha-la into long tubes is a spatially restricted creation of ionic bonds between Ca2+ and carboxyl acid groups on peptide fragments resulting from the action of BLP on alpha-la. Proteolysis of alpha-la with BLP in the presence of Ca2+ thus results in formation of a strong gel with a microstructure not previously observed in food protein systems. PMID- 11504392 TI - Formation kinetics of hydroxymethylfurfural, lactulose and furosine in milk heated under isothermal and non-isothermal conditions. AB - A detailed kinetic study of hydroxymethylfurfural, lactulose and furosine formation was performed upon heating milk at temperatures between 90 degrees C and 140 degrees C. In case of prolonged heating, formation kinetics could be described by a fractional conversion model. Considering only the first phase of the model, kinetics could be simplified to a pseudo-zero order model. A first assessment of kinetic parameters was made by isothermal experiments. Data were analysed using both a 2-step linear and a 1-step non-linear regression method. Only for furosine, did the global 1-step regression approach seem to give better results than the individual 2-step regression approach. Next, the estimated parameters k(ref) and Ea were re-evaluated under non-isothermal conditions by subjecting milk to a time variable temperature profile. Given the complexity of Maillard reaction, it seemed better to estimate kinetic parameters under non isothermal conditions when using a simplified model. Formation of hydroxymethylfurfural, lactulose and furosine was characterized by an Ea value of 90.2 kJ/mol (k(110 degrees C) = 1.2 micromol/l, min), 99.1 kJ/mol (k(110 degrees C) = 51.5 mg/l, min) and 88.7 kJ/mol (k(110 degrees C) = 16.3 mg/100 g protein, min) respectively. Additionally, 90% joint confidence regions were constructed in order to obtain an accurate representation of the statistical confidence associated with the simultaneously estimated parameters. PMID- 11504393 TI - Phenotypic and genetic diversity of enterococci isolated from Italian cheeses. AB - In the present study, 124 enterococcal strains, isolated from traditional Italian cow, goat and buffalo cheeses, were characterized using phenotypic features and randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). The RAPD PCR profiles obtained with four primers and five different amplification conditions were compared by numerical analysis and allowed an inter- and intraspecific differentiation of the isolates. Whole cell protein analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used as a reference method for species identification. The strains were identified as Enterococcus faecalis (82 strains), E. faecium (27 strains), E. durans (nine strains), E. gallinarum (four strains) and E. hirae (two strains). Species recognition by means of RAPD-PCR was in agreement with the SDS-PAGE results except for eight strains of E. faecium that clustered in separated groups. On the other hand, phenotypic identification based on carbohydrate fermentation profiles, using the rapid ID 32 STREP galleries, gave different results from SDS PAGE in 12.1% of the cases. The majority of the strains had weak acidifying and proteolytic activities in milk. One E. faecium strain showed vanA (vancomycin resistance) genotype while four strains showed a beta-haemolytic reaction on human blood. Several strains showed antagonistic activity towards indicator strains of Listeria innocua, Clostridium tyrobutyricam and Propionibacterium freudenreichii subsp. shermanii. PMID- 11504394 TI - Streptococcus thermophilus in cheddar cheese--production and fate of galactose. AB - The behaviour of Streptococcus thermophilus in combination with Lactococcus lactis subsp. cremoris or subsp. lactis mesophilic starters in experimental Cheddar cheese is reported. In a standard manufacturing procedure employing a 38 degrees C cook temperature, even very low levels (0.007%) of Str. thermophilus combined with normal levels of the mesophilic starter (1.7%) resulted in increased rates of acid production, the formation of significant amounts of galactose (approximately 13 mmol/kg cheese), and populations nearly equivalent to those of the mesophilic lactic starter in the curd before salting. At a 41 degrees C cook temperature, the Str. thermophilus attained a higher maximum population (approximately log 8.2 colony forming units (cfu)/g) than the Lc. lactis subsp. cremoris (approximately log 6.8 cfu/g) and formed more galactose (approximately 28 mmol/kg). Lactobacillus rhamnosus, deliberately added to a cheese made using Str. thermophilus starter and which contained 24 mmol galactose/kg at day one, utilized all the galactose during the first 3 months of cheese ripening. Adventitious non-starter lactic acid bacteria had the potential to utilize this substrate too, and a close relationship was demonstrated between the increase in this flora and the disapearance of the galactose. Some possible consequences for cheese quality of using Str. thermophilus as a starter component are discussed. PMID- 11504395 TI - Pinkish colouration in Cheddar cheese--description and factors contributing to its formation. PMID- 11504396 TI - Application of polymerase chain reaction for detection of goats' milk adulteration by milk of cow. PMID- 11504397 TI - Development and application of a quantitative, specific assay for Cryptosporidium parvum oocyst detection in high-turbidity environmental water samples. AB - Chlorine-resistant Cryptosporidium parvum oocysts in drinking water play an important role in the epidemiology of cryptosporidiosis. Current methods of detecting these organisms in water are insensitive, labor-intensive, highly subjective, and severely limited by sample turbidity. We describe here an alternative technique utilizing electrochemiluminescence (ECL) technology for detecting C. parvum oocysts in environmental water samples. This method is quantitative, reproducible, and requires only minimal sample processing. Currently, the ECL assay can detect as few as one oocyst in one milliliter of concentrated test sample with sample turbidity of up to 10,000 nephelometric turbidity units. Water and sewer samples collected during a cryptosporidiosis outbreak were tested by ECL assay. Cryptosporidium parvum oocysts were found in the source water at the time of outbreak, and a sharply decreasing level of oocysts in sewer samples was observed over a three-month period following the outbreak. PMID- 11504398 TI - Differentiation between non-virulent and virulent Burkholderia pseudomallei with monoclonal antibodies to the Ara+ or Ara- biotypes. AB - Burkholderia pseudomallei is the causative agent of melioidosis, a fatal tropical infectious disease endemic in Southeast Asia. Environmental isolates of B. pseudomallei have two distinctive biotypes. Some soil isolates are arabinose assimilators (Ara+ biotype) and are non-virulent in experimental animals. The others cannot assimilate arabinose (Ara- biotype) and are virulent in experimental animals. The Ara- biotype is found in almost all B. pseudomallei clinical isolates. In the present study, a panel of eight monoclonal antibodies that agglutinate the bacteria were produced and tested. The first group, Bps-D2, D3, -D5, -L1, and -L2 agglutinated 100% of Ara+ clinical and soil isolates of B. pseudomallei. Another group Bps-A1, -A2, and -D1 agglutinated 92.9% and 90.9% of Ara- clinical and soil isolates, respectively. This panel of monoclonal antibodies may be useful for rapid differentiation between non-virulent Ara+ and virulent Ara- B. pseudomallei. PMID- 11504399 TI - Short report: characterization of enteroaggregative Escherichia coli isolates from Iranian children. AB - We have previously shown that enteroaggregative Escherichia coli (EAEC) is an important pathogen among Iranian infants and children. To better understand the characteristics of EAEC in Iran, we analyzed EAEC isolates for the presence of pAA plasmid-borne factors. Ninety-eight E. coli strains that displayed the aggregative adherence (AA) pattern on HeLa cells were hybridized with the CVD432 (AA) probe and with genes encoding enteroaggregative heat-stable enterotoxin-1 and aggregative adherence fimbriae (AAF) I and II. Our data suggest that AAF/II is common in this population and that AAF/I and AAF/II can sometimes be detected in the same E. coli isolate. Surprisingly, we have found that AA probe-negative strains in Iran share virulence factors with AA probe-positive isolates and therefore may be more similar to probe-positive strains than previously believed. PMID- 11504400 TI - Protection of pigs with cysticercosis from further infections after treatment with oxfendazole. AB - Cysticercosis, the infection by the larvae of Taenia solium, is a major cause of acquired epilepsy in the world; it also causes significant economic loss because of contaminated pork. This disease is endemic in most developing countries and no control strategy has yet been proven efficient and sustainable. To further evaluate the full potential of single-dose oxfendazole treatment for pigs as a control measure, 20 pigs with cysticercosis were treated with oxfendazole and later matched with 41 naive pigs and exposed to a natural challenge in a hyperendemic area. New infections were found by serologic testing in 15 of the 32 controls (47%), and by the presence of cysts at necropsy in 12 of them (37%). Only minute residual scars were detected in the carcasses of oxfendazole-treated pigs. Pigs with cysticercosis, once treated with oxfendazole, are protected from new infections for at least three months. PMID- 11504401 TI - Short report: floxacrine analog WR 243251 inhibits hematin polymerization. AB - Floxacrine was a promising antimalarial compound that led to the identification of WR 243251. On the basis of their structures, we suspected that these compounds might be good inhibitors of hematin polymerization. Indeed, WR 243251 was as potent and floxacrine was only 2-fold less potent than chloroquine as inhibitors of this process. However, this hematin polymerization inhibition did not completely account for the increased antimalarial potency of WR 243251 versus chloroquine. The WR 243251 ketone hydrolysis product WR 243246 was without activity against hematin polymerization. These data also confirm that hematin polymerization inhibition can be quite sensitive to small changes in inhibitor structure. PMID- 11504402 TI - Plasmodium falciparum dihydrofolate reductase alleles and pyrimethamine use in pregnant Ghanaian women. AB - Drug resistance in Plasmodium falciparum affects prevention of malaria in pregnancy. In a cross-sectional study of 530 pregnant Ghanaian women, P. falciparum dihydrofolate reductase (DHFR) gene mutations linked with pyrimethamine resistance were assessed and associations with pyrimethamine intake were analyzed. P. falciparum infected 69% of women without pyrimethamine use, 59% of those who had a history of pyrimethamine consumption but a negative urine test, and 53% of individuals with a positive urine test. Eighty-one percent, 43%, and 74% of the isolates contained the mutations Asn-108, Ile-51, and Arg-59, respectively. Thr-108 occurred in 8%. Pyrimethamine use was associated with increased frequencies of Asn-108 and Arg-59 but not of Ile-51 or Thr-108. In women with prophylaxis, wild-type parasites were absent and anemia tended to be more common with an increasing number of DHFR gene mutations. Pyrimethamine appears to be not adequately effective in this part of Ghana, most likely due to the predominance of resistant parasites. Selection for resistance following insufficient prophylaxis could possibly affect the efficacy of future intermittent sulfadoxine-pyrimethamine treatment. PMID- 11504403 TI - Assessment of Leishmaniasis notification system in Santiago del Estero, Argentina, 1990-1993. AB - Using a capture-recapture method, this study evaluates the completeness of the cutaneous leishmaniasis (CL) surveillance system in four districts of Santiago del Estero province, Argentina, for the period 1990-1993. Four reporting sources were evaluated: medical records kept by health facilities, interviews conducted during a case-control study, and the national and provincial levels of the leishmaniasis surveillance system (LSS). Using the capture-recapture method it was estimated that 210 cases (95% confidence interval [CI]: 202-218) of CL occurred in the four districts during the study period. Completeness of reporting to the leishmaniasis surveillance system at the national level was estimated to be 44.8% (95% CI: 43.2-46.4). The study results indicate that there is substantial underreporting within the LSS, although it did show the appropriate secular trends. The reasons for under-reporting and methods for addressing this problem are discussed. PMID- 11504404 TI - Short report: transient antibody response in Taenia solium infection in field conditions-a major contributor to high seroprevalence. AB - The discordance between extremely high seroprevalence of Taenia solium antibodies in disease-endemic populations, relatively few symptomatic cases of neurocysticercosis, and high background levels of putatively inactive brain lesions (mainly calcifications) in seronegative controls have confused researchers, clinicians, and epidemiologists in the last decade. We reviewed longitudinal serologic data from general population serosurveys in 3 different disease-endemic areas of Peru and Colombia and found that approximately 40% of seropositive people were seronegative when resampled after 1 year (3 surveys) or after 3 years (1 survey). Transient antibodies may have significant implications for the epidemiology of and immunity to this disease. PMID- 11504405 TI - Longitudinal studies of Sin Nombre virus in deer mouse-dominated ecosystems of Montana. AB - Sin Nombre virus (SNV), hosted by the deer mouse (Peromyscus maniculatus), is the primary etiologic agent of Hantavirus pulmonary syndrome (HPS) in North America. To improve our understanding of the epidemiology of HPS in the western United States, we conducted studies of population dynamics and SNV antibody prevalence in deer mouse populations for 6 years on 12 mark-recapture grids in Montana. Monthly numbers of deer mice ranged from zero to over 170 on 1-hectare grids. SNV antibody prevalence was higher than observed in studies in other parts of the United States, averaging 13% (0% to 50%), and peaking in May or June each year. Antibody-positive mice were older (heavier) (78% of positives were adults versus 52% of negatives) and more likely to be males (61% of positives versus 53.4% of negatives). A higher proportion of antibody-positive deer mice of all age-mass classes had scars than did antibody-negative mice. Month-to-month survivorship of antibody-positive adult mice was similar to that of antibody-negative mice, but survival of young antibody-positive deer mice was lower than antibody-negative deer mice. This is the first study to clearly suggest a detrimental effect of SNV infection on deer mice. PMID- 11504406 TI - Two-dimensional electrophoretic analysis of Aedes aegypti mosquito fat body proteins during a gonotropic cycle. AB - We have used two-dimensional polyacrylamide gel electrophoresis to examine changes in the relative abundance and diversity of non-secreted proteins in Aedes aegypti fat body preparations during a reproductive cycle. Electrophoretic profiles were evaluated at four time points after eclosion, and at five time points after a blood meal. In contrast to the dramatic changes in abundance of specific secreted proteins such as vitellogenin, our results show that the complement of proteins internal to the fat body remains relatively constant during the mosquito reproductive cycle. Of the approximately 5-10% of proteins that do change in abundance, the majority undergo a dramatic decrease within 24 hours after eclosion. PMID- 11504407 TI - Binding of Plasmodium falciparum-infected erythrocytes to soluble platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31): frequent recognition by clinical isolates. AB - Platelet-endothelial cell adhesion molecule-1 or CD31 (PECAM-1/CD31) is a receptor recognized by Plasmodium falciparum-parasitized erythrocytes (pRBCs). Fluorescence-labeled soluble recombinant PECAM-1/CD31 (sPECAM-1/CD31) is shown to bind to the surface of P. falciparum-infected erythrocytes on up to 70% of the cells. Binding is blocked by the addition of the unlabeled receptor in a dose dependent fashion, but not by unrelated receptor-proteins. A significant correlation was found between the binding of sPECAM-1/CD31 to pRBCs and the binding to transfected L cells expressing the receptor as seen with six different P. falciparum lines or clones. Panning of cultures on PECAM-1/CD31 transfected L cells was paralleled by an increase in the binding of sPECAM-1/CD31. The pRBCs of 54% of fresh patient-isolates bound sPECAM-1/CD31 with a mean rate of 12.9% (range = 1.1-44%). The data suggest that PECAM-1/CD31 is a common receptor recognized by wild isolates and that the soluble PECAM-1/CD31 suspension assay is a sensitive and reliable way to study PECAM-1/CD31 binding. PMID- 11504408 TI - Rickettsial infection in murine models activates an early anti-rickettsial effect mediated by NK cells and associated with production of gamma interferon. AB - Natural killer (NK) cell activity was significantly increased on days 2-6 of infection in the Rickettsia conorii-infected C3H/HeN mice and on day 2 in the Rickettsia typhi-infected C57BL/6 mice. Depletion of NK cell activity utilizing anti-NK1.1 monoclonal antibody enhanced the susceptibility of normally resistant C57BL/6 mice to infection with R. typhi, and depletion of NK cell activity with antibody to asialo GM1 enhanced the susceptibility of C3H/HeN mice to infection with R. conorii. Serum gamma interferon was increased in R. conorii-infected C3H/HeN mice compared with NK cell-depleted, infected mice during the early course of infection. Additionally, the NK cell activating cytokine IL-12 was elevated in the sera of infected mice during the time period representing enhanced NK cell activity compared with uninfected mice. Thus, it appears that NK cells contribute to the early anti-rickettsial immune response, likely via a mechanism involving gamma interferon. PMID- 11504409 TI - Sex differences in immune responses and viral shedding following Seoul virus infection in Norway rats. AB - In the field, male rodents are more frequently infected with hantaviruses than females. This study examined whether patterns of immune responses against hantavirus differed between the sexes. Male and female Long Evans rats (Rattus norvegicus) were inoculated with Seoul virus, and antibody and cytokine responses, as well as virus shedding were assessed. Males were more likely to shed virus in saliva, to shed virus through multiple routes (saliva, urine, and feces), and to have viral RNA in the spleen than females. Anti-Seoul virus IgG responses were higher in males than females. In both sexes, splenic IFNgamma and IL-4 production increased following infection. After infection, males had higher Th1 immune responses (i.e., IgG2a, IFNgamma, and IL-2) than females; in contrast, Th2 immune responses (i.e., IgG1, IL-4, and IL-10) were similar between the sexes. These data suggest that immune responses to Seoul virus differ between the sexes. PMID- 11504410 TI - Virulence and viremia characteristics of 1992 epizootic subtype IC Venezuelan equine encephalitis viruses and closely related enzootic subtype ID strains. AB - Following a 19-year hiatus, Venezuelan equine encephalitis (VEE) reemerged in western Venezuela in December 1992. This outbreak is important in understanding VEE emergence because phylogenetic studies imply that sympatric, enzootic, subtype ID VEE viruses mutated to generate the epizootic/epidemic. Although the 1992-1993 strains belong to subtype IC, a serotype implicated in extensive outbreaks during the 1960s and in 1995, relatively small numbers of human and equine cases occurred in 1992-1993. We, therefore, evaluated the pathogenicity of these Venezuelan enzootic ID and epizootic IC viruses to determine 1) if they exhibit phenotypes like those described previously for more distantly related enzootic and epizootic strains, and 2) if the 1992-1993 outbreak was limited by the inability of these IC viruses to exploit equines as amplification hosts. All strains were virulent in mice and guinea pigs, but were benign for cotton rats, natural hosts of enzootic viruses. However, only the IC strains produced equine disease, with mean peak viremias of 10(5) suckling mouse 50% lethal doses per mL serum, and some titers exceeding 10(7). These viremias approximate those observed previously with VEE strains isolated during more extensive epizootics, suggesting that efficient equine amplification did not limit the scope and duration of the 1992-1993 outbreak. Enzootic ID virus infection protected all horses from challenge with epizootic strain P676, supporting the hypothesis that epizootics bypass regions of enzootic transmission due to natural immunization of equines by enzootic VEE viruses. PMID- 11504411 TI - Inflammatory mediators in dengue virus infection in children: interleukin-6 and its relation to C-reactive protein and secretory phospholipase A2. AB - To assess the potential role of interleukin-6 (IL-6) in the pathogenesis of dengue virus infection, levels of this cytokine were measured in children with dengue virus infection on admission to the hospital. As presumed surrogate markers of IL-6, C-reactive protein (CRP) and secretory phospholipase A2 (sPLA2) were measured. Three groups were studied: 33 apparently healthy children as negative controls, 11 children with bacterial infections as positive controls, and 186 children with serologically documented dengue virus infection. One hundred and fifteen patients had dengue fever (DF) and 71 had dengue hemorrhagic fever (DHF). Compared with healthy controls, dengue shock syndrome (DSS) patients had significantly higher levels of IL-6 on admission (P < 0.05), comparable with those in positive controls. Dengue patients with shock had significantly higher levels of IL-6 than normotensive patients (P < 0.001) and higher levels of IL-6 were associated with a higher incidence of ascites. C-reactive protein concentrations in dengue patients and in healthy children were not different, but lower than in children with bacterial infections (P = 0.008). Secretory phospholipase A2 levels were higher in dengue patients than in apparently healthy children (P < or = 0.05) and similar to those in children with bacterial infection. Dengue shock syndrome patients had significantly higher sPLA2 concentrations than normotensive patients (P = 0.02). These data indicate that IL 6 and sPLA2 may have a pathogenetic role only in the most severe forms of dengue virus infection. PMID- 11504412 TI - Case report: recovery from persistent septicemic melioidosis. AB - Septicemic melioidosis is often fatal despite treatment with antibiotics such as ceftazidime to which Burkholderia pseudomallei, the causal pathogen, is sensitive in vitro. We report a near-fatal case of septicemic melioidosis with persistent B. pseudomallei bacteremia despite intravenous ceftazidime in which combination therapy with meropenem and ciprofloxacin, splenectomy and correction of metabolic acidosis allowed for hospital discharge. The choice of antibiotic agents was supported by intracellular minimum inhibitory concentration analysis using B. pseudomallei co-culture in Acanthamoeba trophozoites. The patient's B. pseudomallei isolates were indistinguishable by pulsed-field gel electrophoresis from clinical and environmental isolates previously analyzed during investigation of a Western Australian melioidosis outbreak. A combination of antibiotics known to possess intracellular activity against B. pseudomallei, surgery and supportive critical care may provide a means of improving the probability of survival in persistent septicemic melioidosis. PMID- 11504413 TI - Chronic invasive aspergillosis of the paranasal sinuses in immunocompetent hosts from Saudi Arabia. AB - In immunocompetent patients, paranasal invasive aspergillosis is rare and has a high recurrence rate. Twenty-three cases of paranasal invasive aspergillosis, involving 14 male and nine female immunocompetent patients were reviewed. All patients were cancer-free, HIV-negative, with normal WBC, and none of the patients had received immunosuppressive therapy or corticosteroids. Mean duration of symptoms before diagnosis was 18 months. Aspergillus flavus was the species most frequently isolated. Surgical debridement was performed in all patients followed by antifungal therapy in 18 patients. Mean follow-up duration was 30 months. Fourteen patients relapsed after a mean of 13 months and required an average of 4.3 admissions for repeat surgical evacuation. In a logistic regression model, relapse was not associated with age, duration of symptoms, clinical findings, extent of disease, or mode of therapy. However, patients who were relapse-free tended to have had complete surgical evacuation followed by antifungal therapy. PMID- 11504414 TI - Prevention and control of tropical diseases in the 21st century: back to the field. PMID- 11504415 TI - Construction of SIV/HIV-1 chimeric viruses having the IL-5 gene and determination of their ability to replicate and produce IL-5. AB - During the progression of AIDS, there is a shift in abundance of immune cells from Th1-producing cells to Th2-producing cells. To determine whether this change might have an effect on HIV-1 replication in vivo, we constructed simian/human immunodeficiency chimeric viruses having the human IL-5 gene (a Th2-type cytokine) and examined the effect of the inserted gene on viral replication, IL-5 production and viral stability in vitro. The DNA of human IL-5 was inserted into vpr-deleted and nef-deleted infectious SHIVs. The obtained replication-competent viruses were used to infect human T-cell lines and monkey peripheral blood mononuclear cells. As a result, at the time of peak NI-IL5 virus production, IL-5 was produced with a significantly higher titer than 3sj-IL5. The functionality of the produced IL-5 was confirmed by IL-5-dependent cells. The replication of both SHIVs having IL-5 appeared to be faster than that of the parental viruses without the IL-5 gene. These results show that co-expression of IL-5 stimulates SHIV replication in vitro. Thus, it is expected that expression of IL-5 will also have an effect on viral replication and pathogenicity in vivo. PMID- 11504416 TI - Diverged evolution of recent equine-2 influenza (H3N8) viruses in the Western Hemisphere. AB - We reported previously that equine-2 influenza A virus (H3N8) had evolved into two genetically and antigenically distinct "Eurasian" and "American" lineages. Phylogenetic analysis, using the HA1 gene of more recent American isolates, indicated a further divergence of these viruses into three evolution lineages: A South American lineage, a Kentucky lineage, and a Florida lineage. These multiple evolution pathways were not due to geographic barriers, as viruses from different lineages co-circulated. For the Kentucky lineage, the evolution rate was estimated to be 0.89 amino acid substitutions per year, which agreed with the previously estimated rate of 0.8. For the South American lineage, the evolution rate was estimated to be only 0.27 amino acid substitutions per year. This low evolution rate was probably due to a unique alternating Ser138 to Ala138 substitutions at antigenic site A. For the Kentucky lineage, there was a preference for sequential nonsynonymous substitutions at antigenic site B, which was also a "hot spot" for amino acid substitutions. Convalescent sera had minimal cross-reactivity to viruses of different lineages, indicating antigenic distinctions among these viruses. In contrast to human H3N2 viruses, our results suggested that the evolution of equine-2 influenza virus resembled the multiple evolution pathways of influenza B virus. PMID- 11504418 TI - Investigation of the potential of maize streak virus to act as an infectious gene vector in maize plants. AB - There has been a great deal of interest in the possibility that geminiviruses might be used as infectious gene vectors for expression of foreign proteins in plants. However, generic mastreviruses such as Maize streak virus (MSV) have no sequences which are dispensable for systemic infection of plants, and there is a strict limitation on the size of viral DNA which can be moved systemically. We attempted to complement the movement functions deleted from a wild-type-sized, replication-proficient gene replacement vector, by co-infecting plants with it and either wild type MSV, or a replication-deficient but putatively movement proficient viral construct. While ssDNA formation by the gene replacement vector could be complemented in trans by co-transfected wild type virus, true systemic movement of either the vector, or of co-complementing constructs, did not occur. However, recombination between the two complementing viral constructs frequently occurred to generate wild-type virus genomes. The results therefore suggest that formation of ssDNA and size of the viral replicon are not the sole determinants of whether the MSV movement proteins can mobilise viral sequences and move them systemically in plants. PMID- 11504417 TI - The relative infectivities and genomic characterisation of three distinct mastreviruses from South Africa. AB - The genomic nucleotide sequences of the cloned agroinfectious genomes of three South African mastreviruses obtained from Zea mays, a Setaria sp., and Panicum maximum (designated MSV-Kom, MSV-Set, and PanSV-Kar respectively), were determined. Additionally, their relative infectivities and virulence were analysed in a range of differentially susceptible wheat, maize, and barley genotypes. MSV-Kom produced moderate to severe streak symptoms in all maize genotypes tested, but only moderate to very mild symptoms in the wheat and barley genotypes. MSV-Set infected only the susceptible to tolerant maize genotypes, but was generally more severe in the barley and wheat genotypes than MSV-Kom. PanSV Kar was incapable of infecting any of the wheat and barley genotypes and only produced very mild symptoms on the three most sensitive maize genotypes. Genomic characteristics in common with related mastreviruses were identified. Phylogenetic analysis indicated that while MSV-Kom was closely related to previously sequenced MSV isolates, MSV-Set and PanSV-Kar represented distinctly novel strains of MSV and PanSV respectively. In the case of MSV-Set, this is the most distantly related MSV strain yet characterised. PMID- 11504419 TI - Genetic variation in Australian isolates of myxoma virus: an evolutionary and epidemiological study. AB - Myxoma virus and the European rabbit have been coevolving in Australia since the introduction of the Standard Laboratory Strain (SLS) of myxoma virus as a biological control agent for rabbits in 1950. To examine the degree of genetic variation that has occurred in the virus between 1950 and 1995 and to find genetic markers to use for epidemiological studies, we have examined 37 recent field isolates of myxoma virus for restriction fragment length polymorphisms (RFLPs) by comparision with the progenitor SLS released in 1950. Fifteen RFLPs in Australian isolates have been identified and characterized by mapping and partial DNA sequence analysis. The RFLPs that are most common are deletion and insertion events in regions of tandem repeats. Some of these RFLPs have been investigated by using polymerase chain reaction to identify them in archival tissue samples. In epidemiological studies we characterized two polymorphisms which occur in the Lausanne strain of myxoma virus (widely introduced into Australia from the 1970s) but not in SLS and showed that all of the viruses we isolated were derived from the SLS. PMID- 11504420 TI - Sequence comparison and phylogenetic analysis of fish nodaviruses based on the coat protein gene. AB - We have amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced a 605-bp fragment covering the variable region of the coat protein gene of fish nodaviruses infecting European sea bass, Dicentrarchus labrax (n = 24), and shi drum, Umbrina cirrosa (n = 2), in the Mediterranean basin. Nine new isolates were identified and their sequences were combined with sequences in the literature to produce three different data sets. The first set, based on amino acid sequences, was used to verify the monophyly of fish nodaviruses. The second and third data sets, based on nucleic acids, were used to resolve the phylogenetic relationships between closely related fish nodaviruses. Phylogenetic analyses were performed according to the maximum parsimony and neighbor-joining methods. Our results support the monophyly of fish nodaviruses. Moreover, they confirm the subdivision of fish nodaviruses into four main clusters, in agreement with the previously suggested phylogeny of the genus Piscinodavirus, that was based on a smaller number of sequences and an alternative phylogenetic approach [14]. All the Mediterranean isolates were clustered in the group of the red spotted grouper nervous necrosis virus and appear to have a restricted geographic distribution, except for one sequence-type (10 samples) that is widespread throughout the basin. PMID- 11504421 TI - Plant virus transmission by plasmodiophorid fungi is associated with distinctive transmembrane regions of virus-encoded proteins. AB - Computer analysis of published sequence data has consistently identified two complementary transmembrane domains in the coat protein readthrough domains of benyviruses, furoviruses and pomoviruses and in the P2 proteins of bymoviruses. These viruses differ in genome organisation but are all transmitted by plasmodiophorid fungi. The second domain is absent or disrupted in naturally occurring deletion mutants that cannot be fungally-transmitted. In a non transmissible substitution mutant of Beet necrotic yellow vein virus [Tamada et al. (1996) J Gen Virol 77: 1359-1367], the alignment of the helices is disrupted. From conserved patterns detected in transmembrane helix sequences and calculated relative helix tilts, structural arrangements consistent with tight packing of transmembrane helices were identified. These included ridge/groove arrangements between the two helices and strong electrostatic associations at the interfacial regions of the membrane. The data strongly suggest that these transmembrane helices facilitate the movement of virus particles across the fungal membrane. PMID- 11504422 TI - Rotavirus assembly - interaction of surface protein VP7 with middle layer protein VP6. AB - The interaction between the rotavirus proteins viral protein 6 (VP6) and VP7 was examined in several exogenous protein expression systems. These proteins associated in the absence of other rotaviral proteins as demonstrated by a coimmunoprecipitation assay. Deletion analysis of VP7 indicated that truncations of either the mature amino or carboxyl terminus disrupted the proper folding of the protein and were not able to coimmunoprecipitate VP6. Truncation analysis of VP6 indicated that trimerization of VP6 was necessary, but not sufficient, for VP7 binding. MAb mapping and coimmunoprecipitation interference assays indicate that the VP6 amino acid residues between 271 and 342 are required for VP7 interaction. The interaction of VP6 and VP7 was also examined by the assembly of soluble VP7 onto baculovirus-expressed virus-like particles containing VP2 and VP6. Abrogation of this binding by preincubation of the particles with VP6 MAbs mapped to this same domain of VP6, validated our coimmunoprecipitation results. VP6 IgA MAbs that have been shown to be protective in vivo, but not a nonprotective IgA MAb, can interfere with VP7 binding to VP6. This suggests that these IgA MAbs may protect against rotavirus infection by blocking rotavirus assembly. PMID- 11504423 TI - Population dynamics of a scrapie outbreak. AB - A detailed analysis of a scrapie outbreak in a flock of Cheviot sheep is described. A total of 33 cases of 1473 sheep born to the flock were reported between 1985 and 1994. The epidemiology of scrapie can only be understood with reference to sheep demography, the population genetics of susceptibility to scrapie, pathogenesis during a long incubation period, and the rate of transmission (by both horizontal and vertical routes), all of which interact in complex ways. In recent work a mathematical model incorporating these elements was developed and successfully reproduced key features of an earlier outbreak of scrapie in this flock. Here an application of the model to the second outbreak is described. The model accurately reproduces observed allele frequencies and total numbers of susceptible animals remaining at the end of the outbreak. A major difference between the two outbreaks is the very much lower force of infection in the second outbreak. This provided additional information which suggested two ways in which our existing assumptions be refined; firstly, older animals have reduced susceptibility to scrapie and secondly, homozygous and heterozygous susceptibles have different incubation periods. PMID- 11504424 TI - Characterization of a new adenovirus isolated from black-tailed deer in California. AB - An adenovirus associated with systemic and localized vascular damage was demonstrated by transmission electron microscopy and immunohistochemistry in a newly recognized epizootic hemorrhagic disease in California black-tailed deer. In this study, we describe the cultural, physicochemical and serological characteristics of a virus isolated from lung using neonatal white-tail deer lung and turbinate cell cultures. The virus had the cultural, morphological and physicochemical characteristics of members of the Adenoviridae family. The virus would not replicate in low passage fetal bovine, caprine or ovine cells. Antiserum to the deer adenovirus, strain D94-2569, neutralized bovine adenovirus type-6 (BAdV-6), BAdV-7, and caprine adenovirus type-1 (GAdV-1). Antiserum to BAdV-6 did not neutralize the deer adenovirus but antiserum to BAdV-7 and GAdV-1 neutralized the deer adenovirus. Cross-neutralization with the other bovine, caprine and ovine adenovirus species was not observed. Restriction endonuclease patterns generated for the deer adenovirus were unique compared to those for the currently recognized bovine, caprine and ovine adenovirus types. Amino acid sequence alignments of the hexon gene from the deer adenovirus strain D94-2569 indicate that it is a member of the proposed new genus (Atadenovirus) of the Adenoviridae family. While closely related antigenically to BAdV-7 and GAdV-1, the deer adenovirus appears sufficiently distinct culturally and molecularly to justify consideration as a new adenovirus type. PMID- 11504425 TI - Identification of an immunorelevant ORF2 epitope from porcine circovirus type 2 as a serological marker for experimental and natural infection. AB - Post-weaning multisystemic wasting syndrome (PMWS) is a recently identified disease of pigs linked to the emergence of a new porcine circovirus (PCV2). We report here the characterization of immunorelevant linear B-cell epitopes of the Open Reading Frame 2-encoded protein (Orf2) from PCV2 by an enzyme-linked immunosorbent assay (ELISA) using experimental antisera collected from pigs inoculated with a PCV2 isolate. Two epitopes spanning residues 69 to 83 and 117 to 131 were specific to PCV2. Antibodies to the 117 to 131 epitope (B- 133) were detected in 22% and 100% of specific pathogen-free (SPF) pig sera 6 and 11 weeks post inoculation, respectively. Cross-sectional studies performed with field sera collected from PMWS-affected herds showed B-133 antibodies in 5% of 8 to 10 week old pigs, 38% of 13-14 week-old pigs, 62% of 16 to 19 week-old pigs, 56% of 20 to 25 week-old pigs and 45% of 26 to 31 week-old pigs. All these data suggest that epitope B- 133 is a serological marker of PCV2 infection that could be used for the detection of PCV2 antibody response. PMID- 11504426 TI - Detection of infectious viral particles in plant protoplasts inoculated with transcripts of full-length shallot virus X cDNA. AB - Flexible filamentous shallot virus X (ShVX) particles were detected in extracts of Beta vulgaris protoplasts inoculated with transcripts from a full-length ShVX cDNA. Extracts from ShVX-infected protoplast were infectious for ShVX-healthy shallot seedlings. Western blot analysis of inoculated plants revealed the accumulation of the ShVX coat protein, while electron microscopy confirmed the presence of ShVX virions. The results suggest that the in vitro RNA transcripts from full-length ShVX cDNA give rise to infectious viral particles. PMID- 11504427 TI - Genomic and antigenic variations of the HE glycoprotein of bovine coronaviruses associated with neonatal calf diarrhea and winter dysentery. AB - In this study, we attempted to define differences in the hemagglutinin-esterase (HE) glycoprotein between 11 bovine coronaviruses (BCV) recent (post-1991) and past (pre-1991) isolates from neonatal calf diarrhoea (NCD) and winter dysentery (WD) syndromes as a basis for strain differentiation related to the clinical source of the isolates. The five WD-associated BCV isolates studied could be distinguished from past NCD-isolates by their hemagglutinating (HA) properties at 4 degrees and 37 degrees C, their receptor-destroying enzyme (RDE) activities with rat erythrocytes and lack of reactivity of these NCD isolates to four HA inhibiting (HAI) monoclonal antibodies (MAbs) directed against the HE glycoprotein of the reference WD-associated BCQ.2590 Quebec strain. Although minor or no differences could be demonstrated by comparing biological properties of the HE of WD-isolates to those of recent NCD-isolates, past NCD isolates lacked reactivity with the WD HAI MAbs, whereas recent NCD isolates displayed two distinct reactivity patterns. Attempts to define sequence differences in the HE genes of the WD and NCD strains revealed high nucleotide (NT) identities with only scattered amino acid differences, seemingly unrelated to the clinical origin of the isolates or HAI MAb reactivities. PMID- 11504428 TI - Sequence analysis shows that ribgrass mosaic virus Shanghai isolate (RMV-Sh) is closely related to Youcai mosaic virus. AB - The complete nucleotide sequence of an isolate of Ribgrass mosaic virus (RMV-Sh) from Brassica chinensis (Qingcai) in Shanghai, China was determined. The genome consisted of 6301 nucleotides and its genomic organization was similar to those of other crucifer-tobamoviruses. Comparisons of the nucleotide and predicted amino acid sequences and phylogenetic analyses showed that RMV-Sh had very high homology (> 95% identical nucleotides and 97.7-99.6% identical amino acids) to a sequence of Youcai mosaic virus (YMV = Chinese rape mosaic or Oilseed rape mosaic virus), despite differences in host range or symptoms and this strongly suggests that these isolates should be regarded as belonging to the same species. Only coat protein sequences have been reported for other RMV isolates but it seems likely that the distinction between RMV and YoMV will be difficult to maintain. PMID- 11504429 TI - The Academy of Operative Dentistry recommendations for clinical practice. PMID- 11504430 TI - Fissure caries. PMID- 11504431 TI - Buonocore memorial lecture. Adhesive dentistry applied to the treatment of traumatic dental injuries. AB - Dental adhesive techniques have led to a significant simplification of the immediate and definitive treatment of traumatic dental injuries. Composite restoration of fractured teeth, bonding of tooth fragments, the use of laminate veneers or porcelain onlays and resin retained bridges are some of the ways adhesive techniques are used to treat fractured teeth. Furthermore, splinting of luxated teeth almost entirely relies on the combination of adhesion and a flexible resin which simulates the mobility of a normal periodontal ligament during the healing period. The internal strengthening of immature root-filled teeth with composite using an adhesive technique may possibly prevent late root fractures caused by weakening of the tooth structure resulting from endodontic procedures. Finally, the adhesive principle using a retro-seal with composite after apicoectomy significantly increases the healing rate and healing mode. PMID- 11504432 TI - Pulpal inflammatory responses following non-carious class V restorations. AB - The effects of inflammatory activity following surgical intervention can injure pulp tissues; in severe cases it can lead to pulpal complications. With this article, the authors report on the effects of cavity preparation and restoration events and how they can interact together to reduce or increase the severity of pulpal inflammatory activity in 202 restored Class V cavities. Although some inflammatory activity was observed in the absence of bacteria, the severity of pulpal inflammatory activity was increased when cavity restorations became infected. Zinc oxide eugenol and resin-modified glass ionomer cement prevented bacterial microleakage in cavity restorations, with no severe inflammatory activity observed with these materials. Bacteria were observed in cavities restored with enamel bonding resin and adhesive bonded composites and were associated with severe grades of inflammatory activity. The cavity remaining dentin thickness influenced the grade of inflammatory activity. In the absence of infection, the grade of inflammatory activity decreased after 20 weeks post operatively. In the presence of infection, the grade of pulpal inflammation remained stable until a minimum of 30 weeks had elapsed. PMID- 11504433 TI - Comparative wear resistance of reinforced glass ionomer restorative materials. AB - This study investigated the wear resistance of three restorative reinforced glass ionomer cements (Fuji IX GP FAST [FJ], Miracle Mix [MM] and Ketac Silver [KS]). Microfilled (Silux [SX]) and mini-filled (Z100 [ZO]) composites were used for comparison. Six specimens were made for each material. The specimens were conditioned for one week in distilled water at 37 degrees C and subjected to wear testing at 20 MPa contact stress against SS304 counterbodies using a reciprocal compression-sliding wear instrumentation. Distilled water was used as lubricant. Wear depth (microm) was measured using profilometry every 2,000 cycles up to 10,000 cycles. Results were analyzed using ANOVA/Scheffe's test (p<0.05). After 10,000 cycles of wear testing, ranking was as follows: KS>ZO>MM>FJ>SX. Wear ranged from 26.1 microm for SX to 71.5 microm for KS. The wear resistance of KS was significantly lower than FJ, MM and SX at all wear intervals. Although KS had significantly more wear than ZO at 2,000 to 6,000 cycles, no significant difference in wear was observed between these two materials at 8,000 and 10,000 cycles. Sintering of silver particles to glass ionomer cement (KS) did not appear to improve wear resistance. The simple addition of amalgam alloy to glass ionomer may improve wear resistance but results in poor aesthetics (silver-black color). FJ, which relies on improved chemistry instead of metal fillers, showed comparable wear resistance to the composites evaluated and is tooth-colored. It may serve as a potential substitute for composites in low-stress situations where fluoride release is desirable and aesthetic requirements are not high. PMID- 11504434 TI - Influence of thermal cycling on OCA wear of composite restoratives. AB - This study investigated the effects of thermal cycling on wear of four commercial composite resins (Silux, Z100, Ariston and Surefil). Specimens of each material were divided into three treatment groups comprising a control and two different thermal cycling regimes. Control specimens were stored in distilled water at 35 degrees C for 178 hours. Thermal cycled specimens were stored in distilled water at 35 degrees C for 173 hours and subjected to five hours (300 cycles) of a thermal cycling regime consisting of the cycle ABAC, where A and B represent the fixed temperatures of 35 degrees C (28 seconds) and 15 degrees C (two seconds) and C, depending on the treatment group, was either 45 degrees C or 60 degrees C (two seconds). All specimens were subsequently subjected to wear testing at 20 MPa contact stress against SS304 counterbodies with distilled water as the lubricant. Wear depth (microm; n=6) was measured using profilometry every 2,000 cycles up to 10,000 cycles. Results were analyzed using ANOVA/Scheffe's test (p<0.05). The effect of thermal cycling on wear was material-dependent. The wear of Silux and Z100 were not significantly affected by thermal cycling. Thermal cycling of Ariston at an upper temperature of 60 degrees C significantly decreased wear resistance. Thermal cycling affected only the early wear resistance of Surefil. PMID- 11504435 TI - Marginal quality of tooth-colored restorations in class II cavities after artificial aging. AB - This in vitro study compared the proximal marginal adaptation of direct composite restorations with composite and ceramic inlays inserted with different resin cements. Standardized MOD Class II inlay cavities with one proximal box extending below and the other above the CEJ were cut in 48 extracted human molars and randomly assigned to six groups (n=8). Incrementally layered direct composite restorations (P60), composite inlays (P60) and ceramic inlays (Empress; Cerec Vitablocs Mark II) were placed in the cavities. Three different resin cements (RelyX ARC; Variolink II high viscosity; Panavia 21) were used for luting the composite inlays. All ceramic inlays were cemented with RelyX ARC. After finishing and polishing, the teeth were stored for 24 hours in Ringer solution at 37 degrees C before they were subjected to thermal and mechanical loading (5/55 degrees C, 2000x; 50 N vertical load, 50000x). Margins were evaluated on epoxy replicas using a scanning electron microscope at X200 magnification. Statistical analysis was performed with non-parametric test methods (alpha=0.05). The adhesive interfaces to enamel exhibited high percentages of perfect margins for all groups (91.8% to 96%) and a maximum of 5.2% marginal gap formation. Dentin limited cavity segments demonstrated more marginal openings and less perfect margins than enamel-bound areas; however, this was only statistically significant for direct composite restorations and composite inlays inserted with Variolink II and Panavia 21. RelyXARC showed a significantly better adaptation to P60 inlays compared with the leucite-reinforced Empress ceramic but not the Vitablocs Mark II ceramic. PMID- 11504436 TI - Marginal fit of alumina-and zirconia-based fixed partial dentures produced by a CAD/CAM system. AB - Clinical long-term success of all-ceramic dental restorations can be significantly influenced by marginal discrepancies. As a result, this in vitro study evaluated the marginal fit of alumina- and zirconia-based fixed partial dentures (FPDs) machined by the Precident DCS system. Different master steel models of three-, four- and five-unit posterior FPDs with an 0.8 mm chamfer preparation were produced. FPDs made of DC-Zirkon and In-Ceram Zirconia core ceramics were machined by the Precident DCS system. The marginal fit of the milled frameworks placed on the master steel models was determined by a replica technique using a light-body silicone to fill the discrepancies between crown and tooth and a heavy-body material to stabilize the light-body impression material. The impressions were poured into an epoxy resin material and each model was cross sectioned with a low speed diamond saw to better visualize marginal discrepancies. The marginal fit of the FPDs was evaluated by scanning electron microscopy. The measurements of the marginal fit exhibited mean marginal discrepancies in a range between 60.5 and 74.0 microm, mean marginal gaps in a range from 42.9 to 46.3 microm, mean vertical discrepancies in a range from 20.9 to 48.0 microm and mean horizontal discrepancies in the range of 42.0 to 58.8 microm. Statistical data analysis was performed using the non-parametric test of Kruskal-Wallis and Mann-Whitney. The analysis revealed no significant differences (p>0.05) between the mean marginal gaps and vertical and horizontal discrepancies. However, for some FPDs the mean values of the marginal discrepancies were significantly different (p < or =0.05). The wide range of the measured values may be attributed to the complex geometrical design of long span FPDs and difficulties regarding the milling process of brittle ceramic materials. Based on the selection of 100 microm as the limit of clinical acceptability, the results of this study can conclude that the level of marginal fit for alumina- and zirconia-based FPDs achieved with the Precident DCS system meet the clinical requirements. PMID- 11504437 TI - Effects of different burs on dentin bond strengths of self-etching primer bonding systems. AB - This study evaluated the effects of cutting dentin with different types of burs on tensile bond strength using three self-etching primer bonding systems (Clearfil Liner Bond 2 [LB2], Clearfil Liner Bond 2V [2V] and Clearfil SE BOND [SE], Kuraray, Co, Ltd, Osaka, Japan). Thirty-six intact extracted human third molars were ground flat to expose occlusal dentin, followed by polishing the dentin surfaces with #600 SiC paper. The teeth were divided into four groups according to bur type and grit: fine cut fissure steel bur (SB600), cross cut fissure steel bur (SB703), regular grit diamond bur (DB). Controls were abraded with #600 grit SiC paper (AP#600). The dentin surfaces of the SB600, SB703 and DB groups were cut under copious air-water spray with the respective burs mounted in a dental handpiece. The teeth were treated with one of three adhesive systems, then composite buildups were created with Clearfil AP-X (Kuraray Co, Ltd, Osaka, Japan). After soaking in water at 37 degrees C for 24 hours, serial vertical sections (0.7 mm thick, 7-8 slices per one tooth) were made, trimmed to form an hour-glass shape with a 1.0 mm2 cross-section and tensile bond strengths were determined at a cross-head speed of 1 mm/min. Statistical analysis was made using one and two-way ANOVA and Fisher's PLSD test (p<0.05). Eight additional molars were prepared. Burs or abrasive paper were used for SEM observations of the dentin surfaces of each group before and after treatment with the self-etching primers. All adhesive systems yielded the same ranking of bond strengths to the surfaces prepared with different abrasives: from highest to lowest, AP#600 > SB600 > SB703 > DB. This ranking reached statistical significance using Clearfil Liner Bond 2V (p<0.05). Therefore, when cutting dentin, selecting the adequate bur type is important for improved bonding of adhesive systems using self-etching primer to dentin. PMID- 11504438 TI - Microleakage in amalgam restorations: influence of cavity cleanser solutions and anticariogenic agents. AB - This study evaluated in vitro the influence of five solutions used to treat cavity preparations on microleakage of amalgam restorations. Seventy-two standard Class V cavities were prepared in buccal and lingual surfaces of 36 recently extracted human third molars. The cervical wall was located in cementum and the occlusal wall was located in enamel. Specimens were randomly divided into six groups (n=12) according to the solution employed to treat the cavities: Group I 2.5% sodium hypochlorite; Group II-Calcium hydroxide solution; Group III-1.23% Acidulated phosphate fluoride; Group IV-2% digluconate of chlorhexidine; Group V anionic detergent solution (1.25% sodium lauryl sulfate) and Group VI-control group. The solutions were applied for one minute, followed by washing and drying. Two coats of copal varnish were applied in each cavity. The cavities were filled with capsuled amalgam GS-80 (SDI). After finishing and polishing, the specimens were submitted for thermal cycling followed by immersion in methylene blue. Then, the specimens were sectioned and microleakage was evaluated based on a standard ranking under magnification (40x). Data were subjected to statistical analysis using non-parametric tests. Results of the study concluded that leakage was higher in cementum than enamel (p<0.01). The substances employed have no influence on microleakage except for sodium hypochlorite, which increased leakage values in enamel (p<0.05). PMID- 11504439 TI - Effects of soft-start irradiation on the depth of cure and marginal adaptation to dentin. AB - Marginal adaptation of four resin composites, Clearfil APX, Estelite, Silux Plus and Z-100 cured with two irradiation methods (soft-start or high-power start) of a commercial soft-start halogen lamp unit (Elipar Highlight) were evaluated by measurement of the wall-to-wall contraction gap width. One-hundred and sixty cylindrical cavities, 3 mm in diameter and 1.5 mm in depth, were prepared in extracted human molars. The 80 cavity walls were treated with the Megabond system and each 20 cavities were filled with one of four resin composites. Then, each 10 fillings were irradiated by the soft-start method (soft-power light for 10 seconds followed by high-power light for 30 seconds) or high-power light for 40 seconds. The other 80 cavity walls were treated with an experimental bonding system consisting of 0.5M EDTA as a conditioner, 35% glyceryl mono-methacrylate as a primer and Clearfil Photo Bond as a bonding agent. The cavities were restored wtih the four resin composites and two irradiation methods, the same as the Megabond group. The contraction gap was measured with a light microscope and expressed in % of the cavity diameter. In addition, the curing capability of these two light sources was evaluated by measurement of the curing depth of the four resin composites using a split Teflon mold 4 mm in inner-diameter and 8 mm in height. Marginal gap formation of Clearfil APX, Estelite and Silux Plus with the experimental bonding system was completely prevented regardless of the kind of irradiation methods used. The deterioration of marginal adaptation caused by the Megabond system could not be improved by use of the soft-start method programmed in Elipar Highlight. PMID- 11504440 TI - Adherence of plaque components to different restorative materials. AB - This study compared the amount of artificial plaque synthesized in vitro by Streptococcus sobrinus on various dental materials using radioisotopes. In particular, plaque-retaining capacities of new types of ceramics were the focus of this study. Specimens were fabricated from the following materials (one amalgam alloy [Spherical-D], one casting gold alloy [Casting Gold TYPE I], one resin composite [Herculite XR] and three ceramics [Vita Celay Blanks, IPS Empress and Dicor MGC]). The amount of bacteria and glucans adhered on the specimens was measured after incubation for 24 hours at 37 degrees C with radio-labeled cariogenic bacteria and sucrose. This adhesion test was performed using two different surfaces with 600-grit roughness and clinical smoothness. Irrespective of the surface roughness, the least amount of plaque adhered to the ceramics. However, in the case of the resin composite and amalgam, the amount of bacteria and glucan adhesion decreased dramatically by polishing, though there were no statistically different changes in the amount of bacteria and glucans that adhered to the ceramics even after polishing. In general, the amount of adhered bacteria showed almost the same tendency as that of glucans. Although no statistical differences in the amount of bacteria and glucan adhesion were detected among the three ceramics investigated in this study, a lesser amount of bacteria and glucans adhered to them compared to the other materials. PMID- 11504441 TI - Effectiveness of surface protection of resin modified glass ionomer cements evaluated spectrophotometrically. AB - The effectiveness of four surface protectors for resin-modified glass ionomer cements was evaluated by spectrophotometrically determining dye uptake. Ninety specimens, 3.0 mm in diameter and 1.0 mm in height, were made with Photac-Fil, Fuji II LC and Vitremer and divided into six groups for each material. Positive and negative controls were not protected while experimental specimens were protected with proprietary glaze, nail varnish, flowable resin and glaze. The discs were immersed in 0.1% methylene blue solution for 10 minutes after mixing, except for those negative control specimens that were immersed in deionized water. After 24 hours, the specimens were washed and the protectors trimmed with Sof-Lex discs. The specimens were then removed from the matrixes and individually placed in 1.5 mL of 65% nitric acid for five hours. The absorbance was determined spectrophotometrically at 590 nm. Dye uptake was expressed in microgram dye/specimen. The data were analyzed by two-way ANOVA and Tukey-Kramer tests. All surface protectors tested were effective. For Fuji II LC and Vitremer no differences were observed among tested protections. For Photac-Fil, nail varnish showed better performance than the proprietary glaze. PMID- 11504442 TI - Effect of liners on cusp deflection and gap formation in composite restorations. AB - This study measured deformation of cusps and gap formation associated with MOD resin composite restorations in maxillary premolars with and without the use of low elastic modulus liners. Low elastic modulus liners may reduce the deformation by absorbing polymerization shrinkage stress. Forty maxillary premolars were mounted in stone and slot MOD cavities were prepared. Teeth were randomized into four groups. In Group A, cavities were etched, Single Bond was applied and the cavities were restored with Z-100 composite. In Group B, the same was done except that a layer of flowable composite (Revolution) was place and cured after the bonding agent. In Group C, the same steps were followed as Group A but a layer of glass ionomer (Vitrebond) was placed and cured before the bonding agent. In Group D, a thin layer of composite was placed (after the bonding agent) as a base and cured and the cavities were filled. The distance between indexed cusp tips was measured before the restorations were finished and five minutes and 24 hours after the restorations were completed. The samples were then sectioned mesiodistally and epoxy resin replicas were made and prepared for SEM evaluation of gap formation. The mean contraction of the cusps in microm at 5 minutes and 24 hours, respectively, for each group was A) 47 and 30, B) 35 and 21, C) 23 and 8 and D) 40 and 28. Groups A and D resulted in the highest deformation, B was intermediate and C was the lowest. There was no statistically significant difference in gap formation between the groups. PMID- 11504443 TI - Current teaching of cariology in North American dental schools. AB - All 65 dental schools in the United States and Canada were contacted to obtain information relevant to their cariology teaching effort, the caries management philosophy, the diagnostic threshold for surgical intervention and the non surgical treatment alternatives implemented. Forty-three schools (66%) responded to the request for information. Marked variations in the teaching of cariology and the implementation of modern diagnostic and treatment modalities were noted. The results indicate that with exceptions, efforts to increase curricular time and make organizational changes that would promote cariology as an important academic and clinical endeavor have fallen far short of predictions made more than 20 years ago for North America. Many cariology programs in North American dental schools lack the detail and depth expected for this important area of clinical dentistry. PMID- 11504444 TI - Custom matrix adaptation with elastic cords. AB - Proper placement of the matrix and wedge is critical to the success of proximal amalgam and posterior resin composite restorations. This paper has presented an easy technique for adaptation of the matrix in cases where the gingival cavosurface margin involves a concavity. Special case considerations were also discussed. PMID- 11504445 TI - Physiological levels of melatonin enhance gap junction communication in primary cultures of mouse hepatocytes. AB - Gap junction communication is known to be involved in controlling cell proliferation and differentiation, and seems to play a crucial role in suppression of tumor promotion. Melatonin, a hormone secreted by the pineal gland, has putative oncostatic properties. Intercellular communication through gap junctions was assessed by microinjecting Lucifer yellow fluorescent dye into primary hepatocytes and visualizing the spread of the dye to adjacent neighboring cells using phase contrast/fluorescent microscopy. Treatment of primary hepatocyte cultures with a physiological range of melatonin concentrations for 24 h prior to microinjection resulted in significant enhancement in intercellular communication at 0.2 and 0.4 nmol/L but not at lower (0.1 nmol/L) or higher (0.8 or 1.0 nmol/L) concentrations. A time-dependent study showed that the changes in intercellular communication began 10 h after melatonin treatment and reached a maximum at 12 h of treatment. This nonlinear, functional gap junction response to melatonin occurred in the physiological concentration range detected in blood of mammals during nightly releases of the hormone by the pineal gland. These melatonin levels may affect the ability of gap junction communication to exert cell growth control in vivo. The uneven decline between individuals in nocturnal release of melatonin that occurs with age could identify potentially sensitive subpopulations susceptible to developing pathologies involving alterations in biological processes dependent on gap junction communication. PMID- 11504446 TI - Potential role of antioxidants during ethanol-induced changes in the fatty acid composition and arachidonic acid metabolites in male Wistar rats. AB - Biochemical assessment of liver damage during ethanol-induced stress was done by measuring the activities of serum enzymes, viz., aspartate transaminase (AST) and alkaline phosphatase (ALP), which were significantly elevated in rats fed ethanol. Ethanol administration for a period of 60 days modifies the fatty acid composition, and the analysis of fatty acids showed that there was a significant increase in the concentrations of palmitic acid (16:0), stearic acid (18:0), and oleic acid (18:1) in liver, kidney, and brain, whereas the concentrations of palmitoleic (16:1) and arachidonic acid (20:4) were significantly decreased. The breakdown products of arachidonic acids (20:4), prostaglandins, were elevated. The antioxidants curcumin and N-acetylcysteine (NAC) decreased the activities of serum AST and ALP. Curcumin and NAC decreased the concentrations of fatty acids, viz., palmitic, stearic, and oleic acid, whereas arachidonic acid and palmitoleic acid were elevated. The prostaglandin concentrations were also decreased after curcumin and N-acetylcysteine treatment. Thus the present investigation shows that curcumin and N-acetylcysteine prevent the fatty acid changes produced by ethanol and also reduce the inflammatory response of ethanol by reducing the level of prostaglandins. PMID- 11504447 TI - Toxicity of organotin compounds in primary cultures of rat cortical astrocytes. AB - The neurotoxic organotin compounds trimethyl (TMT) and triethyltin (TET) are known to induce astrogliosis in vivo, which is indicated by an increased synthesis of glial fibrillary acidic protein (GFAP) in astrocytes. In contrast, tributyltin (TBT) does not induce astrogliosis. The aim of this study was to investigate whether trialkyltin derivatives can induce an increased GFAP synthesis in astrocyte cultures in the absence of neurons and whether differences between the action of TMT, TET, and TBT can be detected. Primary cultures of rat cortical astrocytes from 2-day-old rats were grown in 96-well plates until confluency and then exposed to various concentrations of TMT, TET, and TBT for 40 h. Effects on basal cell functions were measured by colorimetric determination of cell protein contents and by assessment of viability by means of the MTT assay. An indirect sandwich ELISA for 96-well plates was used for quantitative measurements of the GFAP content of the cells. All three compounds induced a concentration-dependent cytotoxicity indicated by parallel decreases of protein contents and MTT reduction. Half-maximum cytotoxic concentrations were 3 micromol/L (TBT), 30 micromol/L (TET), and 800 micromol/L (TMT). Cellular GFAP contents were reduced in parallel to cytotoxic action but no increase in GFAP expression at subcytotoxic concentrations could be observed. Thus, the astrocytes were not able to respond to TMT or TET exposure by an increased synthesis of GFAP in the absence of neuronal signals. PMID- 11504448 TI - Induction of lipid peroxidation in hamster organs by the carcinogen cadmium: melioration by melatonin. AB - Cadmium is a well-known human carcinogen. Lipid peroxidation is involved in cadmium-related toxicity and carcinogenesis. Melatonin is an effective antioxidant and free radical scavenger. The potential protective effects of melatonin against cadmium-induced lipid peroxidation in hamster brain, heart, kidney, testes, lung, and liver were examined. Lipid peroxidation was induced by intraperitoneal injection of cadmium chloride [single dose of 1 mg/kg body weight (bw)]. To test whether melatonin would protect against the toxicity of the carcinogen, the melatonin was injected peritoneally at a dose of either 15 mg/kg bw or 5 mg/kg bw, 0.5 h before cadmium treatment and thereafter at 8 h intervals during the day in the 48 h interval following the cadmium injection. One group of hamsters received only a single melatonin injection (a dose of 15 mg/kg bw, 30 min prior to cadmium). Forty-eight hours after cadmium injection, lipid peroxidation increased in brain, heart, kidney, testes, and lung. Either multiple injections of melatonin at both the 5 and 15 mg/kg bw doses, or a single injection of 15 mg/kg bw, prevented the cadmium-related increases in lipid peroxidation in brain, heart and lung. Cadmium-induced lipid peroxidation in kidney was prevented by melatonin when it was given as a single dose of 15 mg/kg bw. Melatonin slightly, but not significantly, reduced cadmium-induced lipid peroxidation in testes. It is concluded that cadmium toxicity, at least with regard to the resulting lipid peroxidation, is reduced by administering melatonin. PMID- 11504449 TI - In vitro effect of lead acetate on human erythropoietic progenitors. AB - Lead is known to induce hematological disturbances resulting from abnormalities in cell differentiation and hemoglobin synthesis during hematopoiesis. The aim of the present work was to study human erythropoiesis in vitro in the presence of lead. Human erythroblastic progenitors, burst-forming units-erythroid (BFU-E), were exposed to lead acetate at increasing concentrations during 14 days of culture. Hematotoxicity was evaluated in vitro according to proliferation and differentiation of cell colonies arising from BFU-E development. The ability of cells to synthesize proteins, porphyrins, and hemoglobin was measured by spectrophotometric tests and by high-pressure liquid chromatography (HPLC). Results showed that in the presence of 10(-3) mol/L lead acetate, no hemoglobinized cells were observed in culture and no fluorescent porphyrins were detected in cells. Up to 10(-3) mol/L, lead acetate is not cytotoxic, i.e., it does not induce cell destruction. The present work demonstrates that lead acetate interferes with the porphyrin synthesis of human erythroblastic progenitors in vitro. The decrease of porphyrin content with 10(-5) mol/L lead acetate suggest that delta-aminolevulinic acid dehydratase can be inhibited by lead acetate during in vitro erythropoiesis. In vivo erythropoiesis occurs in the bone marrow. As about 95% of the body burden of lead in adults is located in the bones with a biological half-life of some years, the concentration of lead acetate found to block porphyrin synthesis in vitro has to be compared with in situ bone marrow lead concentrations. PMID- 11504451 TI - Anti-rotational features for osseointegrated implants. PMID- 11504450 TI - Interaction of benzo[c]phenanthridine and protoberberine alkaloids with animal and yeast cells. AB - We compared the effects of four quaternary benzo[c]phenanthridine alkaloids- chelerythrine, chelilutine, sanguinarine, and sanguilutine--and two quaternary protoberberine alkaloids-berberine and coptisine--on the human cell line HeLa (cervix carcinoma cells) and the yeasts Saccharomyces cerevisiae and Schizosaccharomyces japonicus var. versatilis. The ability of alkaloids to display primary fluorescence, allowed us to record their dynamics and localization in cells. Cytotoxic, anti-microtubular, and anti-actin effects in living cells were studied. In the yeasts, neither microtubules nor cell growth was seriously affected even at the alkaloid concentration of 100 microg/ml. The HeLa cells, however, responded to the toxic effect of alkaloids at concentrations ranging from 1 to 50 microg/ml. IC50 values for individual alkaloids were: sanguinarine IC50 = 0.8 microg/ml, sanguilutine IC50 = 8.3 microg/ml, chelerythrine IC50 = 6.2 microg/ml, chelilutine IC50 = 5.2 microg/ml, coptisine IC50 = 2.6 microg/ml and berberine IC50 > 10.0 microg/ml. In living cells, sanguinarine produced a decrease in microtubule numbers, particularly at the cell periphery, at a concentration of 0.1 microg/ml. The other alkaloids showed a similar effect but at higher concentrations (5-50 microg/ml). The strongest effects of sanguinarine were explained as a consequence of its easy penetration through the cell membrane owing to nonpolar pseudobase formation and to a high degree of molecular planarity. PMID- 11504452 TI - Clinical evaluation of hydroxyapatite-coated single-tooth implants: a 5-year retrospective study. AB - The use of implants to replace missing single teeth has become a valuable treatment option. This study presents a 5-year retrospective clinical analysis of single teeth replaced with hydroxyapatite-coated implants. Of the 271 cylinder and screw implants placed over this period, 12 were lost (survival rate = 95.57%). In addition to aesthetic compromise, the most frequent complication was loosening of the abutment screw. These results corroborate findings from the literature and suggest that the described procedure can be safely performed in daily restorative procedures. PMID- 11504453 TI - Predictable translucency and aesthetic success with pressed ceramics. PMID- 11504454 TI - Use of a synthetic low-fusing quartz glass-ceramic material for the fabrication of metal-ceramic restorations. AB - The development of natural aesthetics has been facilitated by various innovative ceramic materials and techniques. Although metal-ceramic restorations have traditionally been the standard for durable and predictable restorations, the use of metal core materials may result in dense, opaque restorations. The incorporation of low-fusing porcelain materials has enabled the provision of translucent restorations that enable sufficient light transmission and strength. This article presents two comparative case reports that demonstrate the clinical application of a synthetic low-fusing quartz glass-ceramic material for the fabrication of aesthetic and functional metal-ceramic restorations. PMID- 11504455 TI - Material selection--the basis of successful surgical closure. PMID- 11504456 TI - Nickel-titanium rotary instruments: review and strategy for development of a new instrument. PMID- 11504457 TI - Stone models without faces--Part II: An international interview. Clinical Perspectives. Interview by Douglas A. Terry. PMID- 11504458 TI - Digital radiography sensors: CCD, CMOS, and PSP. PMID- 11504459 TI - Cheating yourself...cheating your patients. PMID- 11504460 TI - Full-mouth multidisciplinary restoration using the biological approach: a case report. AB - Bioesthetic dentistry is a conservative approach for the restoration of the teeth to natural form and function. This concept generally requires the examination of the patient's mouth, lips, smile, and face as a single collective structure rather than individual units. Harmonious long-term function depends upon the cohesive relationship between the anterior and posterior dentition, the dentogingival complex, the temporomandibular joints, and the neuromusculature system of the patient. This article defines the role of each of these elements and demonstrates their rehabilitation through a unified multidisciplinary treatment plan. PMID- 11504461 TI - Aesthetic stratification of metal-ceramic crown restorations for natural manipulation of light. PMID- 11504462 TI - EDITORIAL. PMID- 11504463 TI - The IGF/IGF-1 receptor signaling pathway as a potential target for cancer therapy. AB - Expression of insulin-like growth factors (IGFs) and their cognate receptor, the IGF-1 receptor, is frequently upregulated during the development of many types of cancer. Besides stimulating cell cycle progression and the transformation status of tumor cells, a wealth of recent experimental data suggests that IGF-mediated signaling exerts a central tumor-promoting function through the repression of tumor cell apoptosis. These functions are all conveyed by the IGF-1 receptor, thus making it an attractive target for therapeutic intervention. Notably, inhibition of IGF-mediated survival function appears to synergize with conventional chemotherapeutic ablation of tumor cells, raising the possibility of combinatorial cancer therapies with significantly reduced side-effects. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504464 TI - Antifungal resistance in non- albicans Candida species. AB - Non- Candida albicans species have emerged as important bloodstream pathogens. They tend to have decreased susceptibility to antifungal agents in vitro and cause infections associated with high morbidity and mortality. Fluconazole resistance can emerge in any Candida spp., but is most commonly seen with Candida krusei, for which resistance is universal, and with Candida glabrata. Amphotericin B resistance has also been increasingly reported, most notably in isolates of Candida lusitaniae and Candida guilliermondii. Efforts are underway to correlate in-vitro antifungal susceptibility of individual Candida isolates with response to therapy of patients with candidemia. Future advances in this field might allow physicians to identify Candida isolates resistant to specific antifungal agents and thereby tailor therapy of candidemia. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504465 TI - The mechanism of resistance to sulfa drugs in Plasmodium falciparum. AB - The sulfonamide and sulfone (sulfa) group of antimalarials has been used extensively throughout malaria endemic regions of the world to control this important infectious disease of humans. Sulfadoxine is the most extensively used drug of this group of drugs and is usually combined with pyrimethamine (Fansidar), particularly for the control of Plasmodium falciparum, the causative agent of the most lethal form of malaria. Resistance to the sulfadoxine/pyrimethamine combination is widespread. Analysis using molecular, genetic and biochemical approaches has shown that the mechanism of resistance to sulfadoxine involves mutation of dihydropteroate synthase, the enzyme target of this group of drugs. Understanding the mechanism of resistance of P. falciparum to sulfa drugs has allowed detailed analysis of the epidemiology of the spread of drug resistance alleles in the field(1)and, in the future, opens the way to the development of novel antimalarials to this target enzyme. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504466 TI - Activation and role of caspases in chemotherapy-induced apoptosis. AB - The importance of caspase activation during apoptosis has become eminently apparent in the last few years. The caspases participate in a proteolytic cascade activated in response to various stimuli, including anticancer drugs, that results in the systematic and orderly eradication of the cell. The core machinery of caspase activation is now emerging and involves multiple molecular complexes. We describe the two best-studied models of caspase activation, the mitochondrial pathway and the cell death receptor pathway, and discuss their involvement in caspase activation induced by various anticancer drugs used in chemotherapy. Defective apoptosis contributes to tumor growth and drug resistance. Understanding the activation and role of caspases in apoptosis may help develop new therapeutic strategies to circumvent drug resistance. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504467 TI - Causes of drug resistance and novel therapeutic opportunities for the treatment of glioblastoma. AB - Malignant gliomas are among the most lethal and intractable of human tumors and drug resistance is one of the major obstacles to their successful treatment. Recent advances in the molecular biology and genetics of human cancers provide a detailed understanding of cellular and molecular responses to chemotherapy and how drug resistance may develop. Several oncogenes and tumor suppressor genes have been shown to confer resistance to tumor cells and should, therefore, provide novel and defined targets for cancer treatment. In addition to overcoming cellular resistance, special efforts to increase drug delivery to glial tumors need to be pursued because of the relatively unique problem of the blood-brain barrier. Treatments aimed at these targets will likely benefit from combined therapies including surgery, traditional chemotherapy and targeted disruption of other physiological processes such as angiogenesis. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504468 TI - Mechanisms of fluoroquinolone resistance. AB - Mechanisms of bacterial resistance to fluoroquinolones fall into two principal categories, alterations in drug target enzymes and alterations that limit permeation of drug to the target, both resulting from chromosomal mutations. No specific resistance mechanisms of quinolone degradation or modification have been found. The target enzymes, DNA gyrase and topoisomerase IV are most commonly altered in domains near the enzyme active sites and in some cases reduced drug binding affinity has been demonstrated. Drug permeation is altered by mutations that increase expression of endogenous multidrug efflux pumps, alter outer membrane diffusion channels, or both. Recently a new plasmid-mediated resistance of an as yet undefined mechanism was found in clinical isolates of Klebsiella pneumoniae. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504470 TI - Proposed alternatives for the use of anti-HIV drugs. PMID- 11504469 TI - Resistance to non-nucleoside inhibitors of HIV-1 reverse transcriptase. AB - Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are a structurally diverse group of compounds which function as inhibitors of HIV-1 replication in vitro and in vivo. Viral resistance is mediated through alterations in the amino acids which form the allosteric site on the HIV-1 reverse transcriptase to which NNRTIs bind. The rapid emergence of resistant viruses has limited the utility of NNRTI monotherapy; however, recent clinical studies have shown that potent NNRTIs can contribute to profound suppression of HIV replication when used in combination therapy. An understanding of the development of resistance to NNRTIs is critical to the effective use of this class of antiretroviral agents. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504471 TI - Proposed alternatives for the use of anti-HIV drugs: is it appropriate? PMID- 11504473 TI - The TRAIL to an anti-cancer agent. PMID- 11504474 TI - Targeting farnesyltransferase: is Ras relevant? AB - Farnesyltransferase inhibitors (FTIs) are a novel class of cancer therapeutics that were developed to block the localization and thereby the activity of oncogenic Ras protein. Preclinical studies have established that FTIs are nontoxic yet capable of reversing malignant phenotypes. However, there is growing evidence that inhibition of Ras may not be crucial and that the antitransforming properties of FTIs are based at least in part upon alteration of Rho, a small GTPase which is involved in cell adhesion and cytoskeletal regulation. These recent developments are reviewed and their impact on the design of clinical trials is discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504475 TI - The role of CD95 system in chemotherapy. AB - Chemotherapy of malignant tumors, including acute leukemias, is largely based on empirical data and clinical experience. In recent years, it has become clear that anticancer drugs induce apoptosis in target cells. Activation of the CD95 system initiated by induction of CD95 ligand/receptor interaction has been shown to contribute to drug-induced apoptosis in certain systems. However, some groups also have shown that drug-induced apoptosis occurs independently of CD95 triggering. Drug-induced activation of apoptotic pathways appears to include parallel or sequential activation of death receptor systems and, most importantly, changes in mitochondrial function with concomitant release of apoptogenic factors. Activation of downstream caspases, such as caspase 3 or caspase 9, seems to be the most critical mediator of a drug-induced cell death. The identification of the crucial elements of drug-induced apoptosis and activation of effector caspases will certainly provide new insights into the molecular determinants of chemosensitivity or chemoresistance of malignant tumors and leukemias. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504476 TI - HIV resistance to zidovudine: the role of pyrophosphorolysis. AB - Zidovudine-resistant strains of HIV became apparent in many patients soon after advent of zidovudine (AZT) monotherapy. While this resistance could be unequivocally correlated with multiple mutations in HIV reverse transcriptase (D67N, K70R, T215F/Y, K219Q), the mechanism or phenotype for this resistance has remained obscure for more than a decade, despite active investigation. Recent studies indicate that AZT resistance may be related to removal of chain terminating AZT from the 3'-terminus of the primer, by a process known as pyrophosphorolysis. This process is catalyzed by HIV-1 reverse transcriptase (RT), and is the reverse reaction of DNA polymerization. The D67N/K70R mutations result in a significantly increased rate of RT-catalyzed pyrophosphorolysis at physiological levels of pyrophosphate, which leads to a decrease in the extent of AZT chain termination of nascent viral DNA. The potential replication deficit of an increased reverse reaction during DNA synthesis is compensated by increased DNA synthesis processivity, a phenotype that results from the T215F/Y/K219Q mutations in RT. The net result of these multiple phenotypes imparted by the multiple mutations in RT is the facile synthesis of full-length viral DNA in the presence of AZT. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504477 TI - Altered binding of chloroquine to ferriprotoporphyrin IX is the basis for chloroquine resistance. AB - The antimalarial specificity of chloroquine (CQ) stems from the saturable uptake of the drug into malaria parasites. Strains of Plasmodium falciparum that are resistant to CQ have evolved a mechanism to reduce the saturable uptake of CQ and several biochemical models have been proposed to explain this. These include an efflux process analogous to multi-drug resistance (MDR) in cancer cells, reduced proton trapping due to elevated vacuolar pH, reduced binding to an intracellular receptor and reduced activity of a permease or drug importer. Here, we attempt to reconcile many of the apparently conflicting data used to support these models. Previous data are analysed in the context of our own model in which CQ uptake is determined by access of the drug to ferriprotoporphyrin IX (FPIX), the intracellular receptor. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504478 TI - Telomerase targeting in cancer treatment: new developments. AB - Telomerase, a ribonucleoprotein expressed in 85% of advanced cancers but not in most somatic cells, compensates for telomeric DNA erosion and as such stabilizes cell immortality. Telomerase inhibition might restore mortality in tumor cells. Recent progress is illustrated in studies on telomerase and telomere targeting with differentiation induction, reverse transcriptase inhibitors, promoter down regulation, antisense inhibition, and blockage of telomere/telomerase interactions. Also, new developments are described indicating that anti telomerase treatment can induce apoptosis in tumor cells and can chemosensitize drug-resistant cell lines. Implications of these findings for anti-telomerase based therapeutic applications, in particular in combination therapies, are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504479 TI - Antimicrobial peptides from platelets. AB - The fact that platelets play a key role in host defense against infection has been demonstrated by the following observations(1): (a) platelets rapidly respond to sites of endovascular trauma and chemotactic stimuli associated with microbial colonization, and they are the earliest and predominant cells at sites of microbial colonization of vascular endothelium; (b) platelets have surface receptors and cytoplasmic granules comparable in structure and function to those of neutrophils, monocytes, or macrophages; (c) platelets adhere directly to, and may internalize, microbial pathogens, thereby enhancing their clearance from the bloodstream and limiting their potential for hematogenous dissemination; (d) bacterial, fungal, and protozoal pathogens are damaged or killed by activated platelets in vitro; (e) platelets are capable of initiating or amplifying complement fixation in the presence of microorganisms; (f) platelets generate oxygen metabolites which likely contribute to their antimicrobial activity; (g) platelets and leukocytes interact synergistically to exert enhanced antimicrobial functions in vitro; (h) thrombocytopenia increases susceptibility to and severity of certain infections. Importantly, rabbit and human platelets are now known to contain and release microbicidal proteins (termed platelet microbicidal proteins [PMPs] or thrombin-induced PMPs [tPMPs]) when stimulated with microorganisms or platelet agonists associated with infection in vitro. It is hypothesized that these microbicidal peptides accumulate locally at sites of endovascular damage or infection. Recent investigations have confirmed that tPMP-susceptible pathogens are less capable of proliferation or hematogenous dissemination in vivo as compared with their isogenic counterpart strains that are resistant to PMPs. Collectively, the above observations strongly suggest that platelets play key and multi-faceted roles in antimicrobial host defense which appear to be significantly mediated by PMPs and tPMPs. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504480 TI - Recent developments in HIV-1 drug resistance. PMID- 11504481 TI - ATP-binding cassette transporters: from multidrug resistance to genetic disease, Gosau, Austria, 20-27 February 1999. PMID- 11504483 TI - Direct activation of caspases by RGD-peptides may increase drug sensitivity of tumour cells. AB - The realization that cellular homeostasis is dependent on the continuous integration of survival and death signals picked up from the environment and the recent advances in identifying the molecular players involved in these networks may increase our ability to manipulate apoptosis for therapeutic purposes. A recent paper by Buckley et al. in Nature(1)brings this goal one step closer by identifying peptides, containing the motif arginine-glycine-aspartate (RGD), that induce apoptosis by direct activation of caspase-3. We put this finding in the context of what is known about the RGD motif in the light of cancer treatment and suggest the possibility of a synergistic action with anticancer drugs. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504484 TI - GST function in drug and stress response. AB - Glutathione (GSH) is the major non-protein thiol in cells and is a co-factor for many types of conjugation reactions involving glutathione S-transferases (GST). Because of the defined role of GST in drug metabolism, elevated expression of GSTpi either as a tumor marker or in drug resistance has automatically been linked to detoxification. Similarly, GSH has mostly been viewed as a passive cellular supply of nucleophilic sulfur. This mini-review highlights recent reports which advocate a novel, non-catalytic role for GSTpi as an integral determinant of cellular pathways of stress response, proliferation and apoptosis. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504485 TI - Limitations of antifungal agents in the treatment of Candida vaginitis: future challenges. AB - The availability of several potent antifungal agents, systemic or topical, over the counter or prescription would suggest that therapeutic needs for Candida vaginitis are minimal or absent. Unfortunately, unmet needs still exist. Moreover, the pharmaceutical industry has abandoned Candida vaginitis and no new agents or studies are imminent. Perhaps the most important advance in the last decade has been the recognition that therapy must be individualized and that not all forms of Candida vaginitis are equal. A critical factor is duration of therapy and the need for maintenance therapy in recurrent candidiasis. In addition, serious deficiencies exist in the therapy of C. glabrata vaginitis, an emerging problem. Azole therapy for C. glabrata frequently fails, depleting the therapeutic armamentarium of successful options. Additional therapeutic challenges remain for women who can be easily controlled but not cured with intensive azole therapy in spite of absence of in vitro antifungal resistance. Any advance in non-drug related therapy will require a better understanding of the immunopathogenesis of VVC and effective naturally occurring host protective mechanisms. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504486 TI - Glutathione and glutathione-dependent enzymes in cancer drug resistance. AB - Genetic and biochemical evidence has demonstrated that glutathione and glutathione-dependent enzymes play a central role in cellular defence against toxic environmental agents. Modulation of cellular glutathione homeostasis can also have a profound effect on the sensitivity of cancer cells to a wide range of drugs used in chemotherapy. These effects are produced by multifactorial mechanisms that involve inactivation of toxic electrophiles by conjugation, modulation of cellular redox state, activation of drug transporter systems and regulation of cell signalling and repair pathways. New data demonstrating the importance of these pathways in cytoprotection and greater understanding of the mechanisms which regulate their function reveal a number of new targets for novel anti-cancer agents. It is critical, however, if these targets are to be exploited correctly that the dynamics of glutathione regulation are taken into account. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504487 TI - Hsp90 as an anti-cancer target. AB - Heat shock protein 90 is one of the most abundant cellular proteins. Although its functions are still being characterized, it appears to serve as a chaperone for a growing list of cell signaling proteins, including many tyrosine and serine/threonine kinases, involved in cell proliferation and/or survival. The recent discovery of natural products which are able to inhibit Hsp90 function have allowed for both identification of its client proteins and for a better understanding of its role in their activity. Accumulating data have suggested that targeting Hsp90 in cancer cells may be of clinical benefit. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504488 TI - Aminoglycosides for the treatment of gram-negative infections: therapeutic use, resistance and future outlook. PMID- 11504489 TI - Chloroquine - some open questions on its antimalarial mode of action and resistance. AB - During the digestion of its host cell hemoglobin, large amounts of toxic ferriprotoporphyrin IX (FPIX) are generated in the intraerythrocytic malaria parasite. FPIX is detoxified either by being polymerized into hemozoin inside the food vacuole, or through its degradation by glutathione in the cytosol. Chloroquine is able to complex with FPIX, thus inhibiting both processes and thereby generating receptors for its own uptake. These leads to the accumulation of FPIX in the membrane fraction of infected cells that results in membrane permeabilization and disruption of cation homeostasis and concluded in parasite death. Several unresolved questions, such as the site of FPIX:chloroquine complex formation, the role of pH gradient in drug accumulation and resistance, the role of Pgh-1 in resistance, the mode of action of reversers and the involvement of proteins and their mutants in resistance, are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504490 TI - Physiology and molecular genetics of multidrug resistance in Entamoeba histolytica. AB - Entamoeba histolytica presents the evolutionarily conserved multidrug-resistance (MDR) phenotype, discovered in mammalian cells. MDR cells overexpress the membrane P-glycoprotein, which excludes unrelated drugs from the cytoplasm. E. histolytica mutants exhibit cross-resistance to unrelated drugs, which are pumped out from the cytoplasm. In drug-resistant trophozoites, the constitutively expressed EhPg1 gene appears to be up-regulated by a C/EBP-like factor and a multiprotein complex that were not found in drug-sensitive trophozoites. The drug induced EhPgp5 gene, on the other hand, appears to be up-regulated by AP-1 and HOX factors. Here we review the main physiological and molecular facts of the MDR phenotype in E. histolytica. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504491 TI - The 90th Annual Meeting of the American Association for Cancer Research (AACR) Philadelphia, USA, 10-14 April 1999. PMID- 11504492 TI - Regular Article. PMID- 11504493 TI - Thymidine phosphorylase: its role in sensitivity and resistance to anticancer drugs. AB - Thymidine phosphorylase (TP) is an angiogenic enzyme present in normal tissues. Increased levels are found in many tumors, in stromal cells, tumor cells or both. High tumor TP levels may confer a poor prognosis. Cytokines (including interferons), tissue hypoxia and low pH increase TP levels. The influence of tumor TP on fluoropyrimidine toxicity is variable, but capecitabine is a prodrug of fluorouracil that requires activation by TP and hence may have a higher therapeutic index than other fluoropyrimidines. Folate-based thymidylate synthase inhibitors may also be more effective in tumors with a high TP because of increased degradation of endogenous thymidine. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504494 TI - Proteasome inhibitors as potential novel anticancer agents. AB - Ubiquitin/proteasome-dependent proteolysis plays an essential role in degrading regulatory proteins and thereby controlling processes of cell proliferation and cell death (apoptosis). Most recent experiments using cell cultures and mouse models have demonstrated that proteasome inhibitors induce cancer cell apoptosis and therefore inhibit tumor growth. The proteasome inhibitors have the following unique features: (i) greater apoptosis-inducing potency when tested in various human tumor cell lines than current anticancer drugs; (ii) ability to selectively target transformed and tumor, but not normal, human cells; and (iii) ability to overcome tumor cell resistance to cytotoxic therapies. We suggest that proteasome inhibitors have potential use as novel anticancer agents. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504495 TI - Vancomycin-resistant enterococci: recent advances in genetics, epidemiology and therapeutic options. AB - Vancomycin-resistant enterococci (VRE) have gained much attention in the last decade. Currently, there are five known types of vancomycin resistance based on genes encoding ligase enzymes that the organisms use to produce their cell wall precursors, namely, VanA, VanB, VanC, VanD and VanE. An additional unclassified type was discovered in Australia. The basis of resistance among these phenotypes appears to be similar in that the resistant organisms produce peptidoglycan precursors that end in moieties other than D-alanyl-D-alanine, the usual target of vancomycin. The other dipeptide-like termini identified to date include D alanyl-D-lactate and D-alanyl-D-serine, which have low affinity for glycopeptides. Recent evidence suggests that glycopeptide-producing organisms might be the remote origin of the vancomycin resistance genes. In European countries, avoparcin, a glycopeptide used in farm animals as a growth promoter, has been linked to the occurrence of VRE and occasional common strains have been identified in food products, farm animals, healthy subjects and hospitalized patients. There have been no such reports in the USA where heavy use of vancomycin and use of broad spectrum antibiotics such as cephalosporins have been identified as important risk factors for acquisition of VRE. Transmission within the same or between hospitals has been reported in many countries. Infection control measures and efforts to use antibiotics, particularly vancomycin, more appropriately have been implemented in a number of healthcare facilities with varying degrees of success. Many antibiotics, as a single agent or a combination of drugs, as well as various new antibiotics have been tested in vitro, in animal models, or used in anecdotal cases but clinical data from large comparative trials are not available to date. Because of the limited susceptibility of many VRE to other agents, efforts to control these organisms are particularly important. Copyright 1999 Harcourt Publishers LtdCopyright 1999 Harcourt Publishers Ltd. PMID- 11504496 TI - Chemotherapy of respiratory viruses: prospects and challenges. AB - Billions of people are infected with respiratory viruses annually. Infants and young children, the elderly, immunocompromised individuals and those debilitated by other diseases or nutritional deficiencies are most at risk for serious disease. There are few vaccines available for use against these viruses, and even where there are (influenza, measles and adenovirus), infections remain common. The continued prevalence of respiratory virus infections has lead to renewed efforts to find safe agents effective against the most medically important respiratory viruses: influenza, respiratory syncytial, parainfluenza, measles, rhino- and adenovirus. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504497 TI - Drug resistance in Cryptococcus neoformans. AB - Cryptococcus neoformans has become a major opportunistic fungal pathogen worldwide. Successful treatment of invasive disease with this fungus has used amphotericin B, flucytosine and various azoles. However, treatment failures continue to occur for a variety of reasons including direct antifungal drug resistance. Issues and mechanisms for antifungal drug resistance in Cryptococcus neoformans are reviewed. Furthermore, approaches and strategies for prevention and treatment of antifungal drug resistance are identified and these include host immune modulation, dose optimization, prophylaxis/empirical regimens, improved drug delivery systems such as lipid preparations of amphotericin B, surgery, combination antifungal treatments and development of new antifungal agents. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504498 TI - NF- kappa B and chemoresistance: potentiation of cancer drugs via inhibition of NF- kappa B. PMID- 11504499 TI - NF- kappa B and chemoresistance: could NF- kappa B be an antitumor target? PMID- 11504501 TI - Small cell and its stroma: determinant of pan-resistance? PMID- 11504502 TI - The tumor microenvironment as a determinant of drug response and resistance. AB - Classically, studies of drug resistance in cancer have focused on the molecular biology of single cancer cells. These types of studies have provided important information regarding certain drug resistance mechanisms, including mechanisms that reduce intracellular drug accumulation, alter or repair drug-induced damage, and reduce drug-induced apoptosis. While these cellular mechanisms undoubtedly contribute to the overall phenomenon of drug resistance, it is now evident that the tumor cell microenvironment also influences how a tumor cell behaves and responds to cytotoxic drugs or radiation. Two different forms of tumor cell environmental interaction may explain how some tumor cells survive initial drug exposure and eventually express classical mechanisms of drug resistance. The first form involves soluble mediators, such as interleukins, that are secreted by non-tumor, stromal cells. Interleukin-6 (IL-6) is a classical example of how a soluble mediator secreted by the tumor microenvironment is capable of enhancing tumor cell survival and perhaps blocking apoptosis. The second form of tumor cell environment interaction requires direct cell contact and has been given the term cell-adhesion-mediated drug resistance (CAM-DR). In this case, binding extracellular matrix ligands in the tumor microenvironment may activate cell adhesion molecules, such as the integrins, and these interactions result in the activation of signal transduction pathways that block drug-induced apoptosis. Interrupting the tumor cell-environment interactions or the associated signal transduction pathways may represent a new approach for the treatment of cancer. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504503 TI - Molecular basis of metronidazole resistance in pathogenic bacteria and protozoa. AB - The molecular basis of metronidazole resistance has been examined in anaerobic bacteria, such as Bacteroides, Clostridium, and Helicobacter, and anaerobic parasitic protists such as Giardia, Entamoeba, and trichomonads. A variety of enzymatic and cellular alterations have been shown to correlate with metronidazole susceptibility in these pathogens; however, a common theme has been revealed. Resistant cells are typically deficient in drug activation. The frequent correlation between metronidazole resistance and ineffective drug activation suggests that drug resistance is the result of modification of proteins involved in drug activation. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504504 TI - Mismatch repair and drug responses in cancer. AB - Defects in mismatch repair contribute to development of approximately 15% of colon cancers and to origination of endometrial, gastric and other cancers. Tumors with defects in mismatch repair exhibit marked resistance to alkylators and a variety of anticancer agents that modify DNA to create substrates for the mismatch repair system. These altered drug responses appear to derive from requirements for mismatch repair proteins in signalling apoptosis, altered cell cycle checkpoint behaviour and/or loss of mismatch repair dependent toxicity arising from futile repair cycling. Altered repair mechanisms for mismatched substrates in mismatch repair defective tumors provide both challenges for development of tumor-phenotype-screening methodologies to assure appropriate therapy is administered for these cancers and foci for development of new therapy approaches that capitalize on modified drug responses in mismatch repair- defective cells. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504505 TI - Topoisomerase I inhibitors: selectivity and cellular resistance. AB - Topoisomerase I (top1) inhibitors (camptothecins and other structurally diverse compounds) are effective and promising anticancer agents. Determinants of selectivity toward cancer cells and resistance are multifactorial. These factors can be separated in three groups. The first is related to alterations in drug distribution and metabolism. The second group includes both quantitative and qualitative (mutations) differences in top I. The third group includes resistance and sensitivity factors downstream from the cleavage complexes. They include DNA repair, cell cycle checkpoints and apoptosis, and are probably key to the relative selectivity of camptothecins toward cancer cells and to clinical resistance. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504506 TI - Cell wall targets in methicillin-resistant staphylococci. AB - Multiresistant staphylococci pose an alarmingly growing problem, especially in serious hospital infections. The recent emergence of strains with reduced susceptibility against vancomycin, the last remaining drug effective against methicillin (multi) resistant Staphylococcus aureus, highlights the urgent need for new antimicrobial agents and new therapeutic regimen. Previously, new drugs were discovered exclusively in bacterial whole cell growth assays. Today's more rational approach depends on the identification of suitable target genes and proteins. These should be bacteria-specific and essential for growth either in vitro or in vivo. Targets within cell wall synthesis and remodeling pathways might be particularly attractive because the bacterial cell wall is a unique structure occurring only in prokaryots; many of the antibiotics in use today have confirmed its 'drugability'. However, several potential targets within this field have not yet been exploited successfully for anti-staphylococcal therapy and some were discovered only recently. After a short summary of known potential targets a set of genes involved in the pentaglycine interpeptide bridge formation of the staphylococcal cell wall will be introduced as interesting targets to combat multiresistant staphylococcal infections. Copyright 1999 Harcourt Publishers LtdCopyright DUMMY. PMID- 11504507 TI - Antifungal drug susceptibility testing and resistance in Aspergillus. AB - Aspergillus species are the most common causes of invasive mold infections in immunocompromised patients. The introduction of new antifungal agents, and recent reports of resistance emerging during treatment of aspergillus infections, have highlighted the need for standardized methods of antifungal drug susceptibility testing for filamentous fungi. This review describes the methods that are now being developed for the in vitro testing of Aspergillus species, and the results of attempts to correlate in vitro findings with in vivo outcome. The mechanisms and clinical importance of resistance to the different agents used in the treatment of human aspergillosis are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 11504508 TI - 39(th)Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC) San Francisco, USA, 26-29 September 1999. PMID- 11504510 TI - The rectal cancer debate. PMID- 11504511 TI - The role of total mesorectal excision in rectal cancer surgery. PMID- 11504512 TI - Is radiotherapy for rectal cancer indicated if surgery is optimized? PMID- 11504513 TI - Recent pathology related advances in colorectal adenocarcinomas. AB - This brief review addresses some new developments in the understanding of the molecular pathogenesis of colorectal cancer, including the mismatch repair protein mechanism of colorectal adenocarcinoma carcinogenesis. The second part of this review summarizes recent improvements in the pathological evaluation of colorectal resection specimens, and highlights the advantage seen in the resulting improved quality of gross dissection provided by the use of highly trained non-medical pathology assistants. Topics also covered include optimum methods of assessment of radial resection margins, the accuracy of measured margin distances, the assessment of adenocarcinoma involvement of the peritoneal serosal surface, improved methods of lymph-node recovery from colorectal adenocarcinoma resection specimens and the recently recognized adverse prognostic significance of extranodal pericolonic tumour deposits. Formats for standardized reporting of colorectal resection specimens are presented as a means to improve quality and consistency of pathological data recording and collection. PMID- 11504514 TI - Prognostic factors for gastric cancer in the elderly. PMID- 11504515 TI - Aggressive local treatment for screen-detected DCIS results in very low rates of recurrence. AB - AIMS: To review our institution's practice of treatment of a mammographically detected population of ductal carcinoma in situ (DCIS) patients and to determine the outcome. METHODS: Between April 1989 and March 1994, 304 women with median age 59 years (range 51-65) with DCIS detected on screening mammogram, were treated in the Newcastle General and Royal Victoria Infirmary Hospitals, Newcastle-upon-Tyne, UK. More than half of the women (n=176, 57.8%) decided to have mastectomy. Other treatment options were wide local excision (WLE) with radiotherapy (n=97, 32%) and WLE alone (n=31, 10.2%). All except five received adjuvant hormone treatment. RESULTS: Predominant DCIS was comedo in 122 (42%), followed by cribriform in 87 (30%) and micropapillary in 44 (15%) cases. Grade I was found to be commonest grade (54%) followed by grade II (27%) and grade III (11%). With a median follow-up of 88 months, there were six (2%) recurrences, all of which were in women who were given breast conservation treatment, WLE with radiotherapy (n=1, 1%) and without radiotherapy (n=5, 16.6%). Mastectomy in this series was not associated with any recurrence at all. In three cases the recurrence was invasive, one of who also had distant metastasis. CONCLUSIONS: The findings of this study suggest that in women with DCIS suitable for breast conservation, WLE when combined with radiotherapy is associated with a very low recurrence rate. PMID- 11504516 TI - Pathological features of mucinous carcinoma of the breast are favourable for breast-conserving therapy. AB - AIM: The effectiveness of breast-conserving therapy for mucinous carcinoma has not been well documented. We examined clinical and pathological features of cases to determine whether patients with mucinous carcinoma were suitable candidates for this treatment. METHOD: Cases of pure type (n=52) and mixed type (n=24) mucinous carcinomas were reviewed with emphasis on the risk factors associated with local recurrences after breast-conserving therapy. RESULTS: Large pure mucinous carcinomas had a low incidence of extensive intraductal spreading (EIS). An inverse correlation existed between the incidence of EIS and tumour size (P<0.05). Comedo type EIS was infrequent (11%) in pure mucinous carcinoma. Incidences of lymphatic vessel invasion (4%) and nodal involvement (4%) were lower in pure mucinous carcinoma than in mixed carcinoma (P<0.05). No nodal involvement occurred in patients with pure mucinous carcinoma less than 3 cm in diameter. CONCLUSIONS: Patients with pure mucinous carcinomas, except those invading the local skin, are suitable candidates for breast-conserving therapy. Most pure mucinous carcinomas, including a large tumour up to 5 cm in diameter, can be treated with this therapy. PMID- 11504517 TI - Cyclin E overexpression as an independent risk factor of visceral relapse in breast cancer. AB - AIM: Prognostic value of the cyclin E overexpression in breast cancer has not been clearly established, especially in relation to the pattern of recurrence. We investigated the implication of cyclin E overexpression for the pattern of recurrence in Korean breast cancer patients. METHODS: Using immunohistochemical methods, we retrospectively examined the cyclin E expression level in breast cancer specimens from 128 women who underwent curative breast surgery, and correlated the levels of expression with the pattern of relapse in patients. RESULTS: Cox model-based multivariate analysis indicated that distant relapse could be predicted by the number of positive axillary lymph nodes, high cyclin E expression, and the younger age (<35 years) of the patient. We tested further the association of cyclin E overexpression with the specific types of recurrence; multivariate analyses indicated that adjusted relative risks of bone and visceral relapse as the first events among high cyclin E group were 2.46 (95% confidence interval (CI), 0.86-7.02) (P=0.092), and 3.98 (95% CI, 1.23-12.94) (P=0.022), respectively. On the other hand, cyclin E overexpression was not associated with the risk of locoregional relapse. CONCLUSION: Our data suggest that cyclin E overexpression in primary breast carcinoma tissue could independently predict the risk of distant relapse, especially of visceral relapse, as the first failure after curative breast surgery. PMID- 11504518 TI - Thrombin enhances adhesion in pancreatic cancer in vitro through the activation of the thrombin receptor PAR 1. AB - BACKGROUND: Thrombin, the central enzyme of the coagulation cascade, induces proliferation in different solid tumours. The effect is mainly mediated through the functional thrombin receptor PAR 1, a member of the G-protein coupled receptor family. The aim of this study was to assess the role of thrombin on adhesion of pancreatic cancer to extracellular matrix proteins and endothelial cells in vitro. MATERIALS AND METHODS: The human pancreatic adenocarcinoma cell line MIA PaCa-2 was treated with thrombin and the thrombin-receptor-activating peptide (TRAP), respectively. As a control the cells were pre-incubated with the thrombin-receptor-inhibiting peptide (T1). The cells were incubated on microtiter plates, which were pre-coated with extracellular matrix proteins (fibronectin, laminin, collagen IV) or human umbilical vein endothelial cells (HUVECs), for 30 and 60 min, respectively. The number of adherent cells were measured using the MTT method. ANOVA was used for statistical analysis. RESULTS: Thrombin enhanced the adhesion of MIA PaCa-2 cells to extra-cellular matrix proteins and endothelial cells significantly (P< or =0.001). The effects of thrombin could be mimicked by TRAP. Pre-incubation with T1 inhibited the effect. CONCLUSION: Thrombin enhances adhesion of pancreatic adenocarcinoma to extracellular matrix proteins and endothelial cells in vitro. The effect is mediated through the thrombin receptor PAR 1. The results emphasize the role of thrombin and PAR 1 in pancreatic tumour biology. PMID- 11504519 TI - Treatment of non-resectable pancreatic cancer with intraperitoneal 5-FU and leucovorin IV. AB - AIM: To explore the feasibility of intraperitoneal (IP) 5-fluorouracil (5-FU) and (IV) leucovorin for patients with advanced pancreatic carcinoma. MATERIALS AND METHOD: Thirty patients (11 men), median age 65 (range 36-74 years), with a non resectable pancreatic carcinoma in stage III (n=2) and IV (n=28) were treated with IP 5-FU 750-1000 mg/m(2)and leucovorin IV 100 mg/m(2)for 2 days every 3rd week. Tumour effect was analysed with repeated computed tomography (CT) scans, performance status was estimated with Karnofsky's index (KI) and morphine consumption, and toxicity assessed using World Health Organization (WHO) criteria. RESULTS: Median survival time was 7 months (range 0-21). There was no difference in survival between patients with different grading, staging or tumour size. Regional and systemic toxicity: The treatment was well tolerated, with no grade III or IV complications or side-effects. The median KI showed a minor reduction during treatment. The median morphine consumption per 24 hours increased from 0 mg (range 0-250) at inclusion, to 70 mg (range 0-540) at exclusion. The median nadir (WBC) was 7.2x10(3)/mm(3)(range 5.2-18.8). All patients had abdominal discomfort and distension during IP installation. CONCLUSION: Intraperitoneal administration of 5-FU is feasible for patients with nonresectable pancreatic carcinoma. The treatment can induce a temporary stabilization of tumour growth and eventually prolong survival without adverse effects. PMID- 11504520 TI - Hydronephrosis associated with colorectal carcinoma: treatment and outcome. AB - AIM: Obstruction of the upper urinary tract, hydronephrosis, is not uncommon in the context of primary or recurrent colorectal cancer (CRC). Its presence poses a therapeutic dilemma. This study focuses on the significance of hydronephrosis as a prognostic marker for CRC by analysing the resectability and survival rates of patients affected. PATIENTS AND METHODS: Retrospective data of 52 patients with hydronephrosis were analysed. Ten had primary CRC at different sites and 42 developed hydronephrosis 1-84 months following resection of a primary CRC. Twenty eight had unilateral and 24 bilateral hydronephrosis. RESULTS: In 10 patients with primary CRC and in 38 of those with a history of CRC, hydronephrosis was secondary to malignant obstruction. In four it was related to iatrogenic injury to the urinary tract. Complete surgical resection was possible in five patients (10%) with malignant obstruction. The remaining 90% underwent palliative or no surgical treatment due to diffuse metastasis or extensive local disease. No difference in survival was found between these two groups (6 vs 8 months) nor when comparing CEA levels, Duke's staging, or unilateral vs bilateral hydronephrosis. Patients with benign obstruction were treated by a ureteric stent, leading to resolution of hydronephrosis. All four are alive. CONCLUSIONS: Malignant hydronephrosis, secondary to primary or recurrent CRC, represents local manifestation of a disseminated disease with almost no probability of long-term survival and cure. It would seem that patients with such disease do not benefit from aggressive operations. PMID- 11504521 TI - Is MRI more accurate than CT in estimating the real size of adrenal tumours? AB - BACKGROUND: The size of adrenal tumour plays an important role in the indications for surgical excision of non-functioning adrenal tumours and in selecting the best surgical approach. Computed tomography (CT) has been reported to underestimate the real size of adrenal lesions. The accuracy of magnetic resonance imaging (MRI) in predicting the true tumour size has not been previously investigated. The present retrospective study investigates the accuracy of MRI and CT in the pre-operative determination of true adrenal tumour size. METHODS: The medical records of 65 patients who underwent adrenalectomy for an adrenal mass were reviewed. The size of adrenal tumours as determined by pre operative MRI and/or CT was compared with the "true" histopathological size. The impact of histological diagnosis on size estimation was also investigated. RESULTS: The median age at diagnosis was 42 years (range 1-82 years) and more patients were female (60%). Five patients had bilateral adrenalectomy, thus giving rise to 70 adrenal specimens. The histopathological size of adrenal tumours ranged from 0.9 to 26 cm with a mean of 5.96 cm and a median of 4.70 cm. For tumours larger than 3 cm, MRI significantly underestimated the real tumour size by 20% (P<0.001). CT also underestimated the size of such tumours by 18.1% (P<0.003). Adrenal phaeochromocytomas were consistently underestimated by both modalities. CONCLUSIONS: MRI and CT significantly underestimated the true size of adrenal tumours larger than 3 cm by 20% and 18%, respectively. Surgeons and endocrinologists should interpret the pre-operative size of adrenal lesions with caution. PMID- 11504522 TI - The management of retroperitoneal soft tissue sarcoma: a single institution experience with a review of the literature. AB - AIM: Ten percent of soft tissue sarcomas (STS) arise in the retroperitoneal tissues. The prognosis for patients with retroperitoneal sarcoma is poor with a 5 year survival rate between 12% and 70%. Stage at presentation, high histological grade, unresectable primary tumour and incomplete resection are associated with a less favourable outcome. METHODS: Complete follow-up data were available on 22 patients who underwent surgery for retroperitoneal STS in our institution between 1990 and 2000. Patient, tumour and treatment variables were analysed including use of adjuvant therapy and survival status. RESULTS: Eighteen patients underwent surgery for primary disease, four patients were treated for recurrent disease or metastases. Ten patients presented with pain, seven with an abdominal mass, other presentation included weight loss and haematuria. Thirteen patients presented with tumours larger than 10 cm. The tumours were seven liposarcomas, six leiomyosarcomas, three malignant fibrous histiocytomas, two rhabdomyosarcomas, two malignant schwannomas and two undifferentiated sarcomas. Six primary tumours were completely excised, five patients received radiotherapy and five received chemotherapy. Local recurrence rate was 45% and recurrence-free interval for 10 patients with recurrence was 11 months. Five patients received radiotherapy and five received chemotherapy. The median survival for patients with primary tumours was 36 months, and 5-year survival was 44%. Adjuvant therapy was not associated with higher survival rates. CONCLUSION: This study re-emphasizes the poor outcome of patients with retroperitoneal STS. Adjuvant radiotherapy and chemotherapy do not appear to be any proven benefit and the single most important prognostic factor is aggressive successful en bloc resection of the primary tumour. Our resection rate and 5-year survival rates are comparable with previous reported UK series although lower than large reports from North American centres. This might partly be explained by difficulty in data collection in a retrospective analysis, but may reflect inadequate subspecialization in UK centres. PMID- 11504523 TI - Pathology of ductal carcinoma in situ of the breast: current status. AB - Ductal carcinoma in situ (DCIS) now accounts for around 20-25% of mammographically detected breast cancers. There is strong evidence to show that classification schemes for DCIS should be based primarily on nuclear grade and necrosis as these two features have been shown to be prognostically important as well as having high interobserver reproducibility among pathologists. Newer classifications of DCIS that employ these features, such as the Van Nuys DCIS Classification, are of prognostic importance in predicting recurrence of DCIS after breast conservation and show high levels of reproducibility. For treatment of DCIS via breast conservation a high pre-operative diagnostic rate is desirable, only achievable via needle-guided core biopsy. If local excision without radiotherapy is to be given there is strong evidence to support the requirement for a 10 mm tumour-free margin. Assurance that a margin is tumour free requires sequential specimen processing which is both time consuming and costly, but which can be justified in cost and morbidity terms as radiotherapy may not be required for those patients with a 10 mm tumour-free margin. Other methods of specimen examination involve examination of mammographically directed tissue slices or alternative methods of excision margin assessment such as "onion skinning" of the specimen. Endocrine therapy will doubtless become more important for adjuvant therapy of DCIS as well as chemoprophylaxis in the future. PMID- 11504524 TI - DNA microarray technology in cancer research. AB - Microarray technology transforms the study of functional genetics. The entire genomic activity of cells and tissues can be analysed and compared on single slides, or gene chips. In cancer research, this will allow the better understanding of the regulation of activity of cells and tumours in various states. It will also allow the classification of individual tumours by their gene expression patterns, which may also describe and predict therapeutic resistance and sensitivity patterns. This short article provides a short introduction to the technology and its applications. PMID- 11504525 TI - Long-term survival of a patient with widespread metastases from Barrett's adenocarcinoma. AB - Five-year survival of patients with stage 4 Barrett's adenocarcinoma is minimal. Such a patient is presented, alive without evident disease, 7 years after resection of the primary lesion, during which time he drank Peruvian herbal tea. PMID- 11504526 TI - Extensive cerebral invasion of a basal cell carcinoma of the scalp. AB - Basal cell carcinomas (BCC) are the most common cutaneous malignant neoplasms affecting fair-skinned human beings. They may progressively enlarge and spread by peripheral extension. Intracranial invasion is very rare. A case of a giant BCC of the scalp invading directly into the left hemisphere through the frontal and parietal lobe and meninges is presented. Brain invasion along Virchow-Robin's spaces is demonstrated histopathologically. A review of literature failed to disclose a previous report of similar extensive and fatal cerebral invasion from a basal cell carcinoma of the scalp. PMID- 11504527 TI - Breast cancer in two sisters with Friedreich's ataxia. AB - Friedreich's ataxia (FRDA) is an autosomal recessive neurodegenerative disorder causing progressive ataxia and dysarthria. We report two sisters who had breast cancer aged 39 years and 42 years and who both developed a late onset form of FRDA with onset of neurological symptoms in their thirties. We discuss whether there may be an association between the late onset form of FRDA and malignancy. PMID- 11504528 TI - A.D. Jenkinson et al. Does intraductal breast cancer spread in a segmental distribution? Eur J Surg Oncol 2001; 27:21-25. PMID- 11504529 TI - Zang RY, Zhang ZY, Li ZT et al. Impact of secondary cytoreductive surgery on survival of patients with advanced epithelial ovarian cancer. Eur J Surg Oncol 2000; 26:798-804. PMID- 11504530 TI - Carbohydrate and fat metabolism and related hormonal regulation in normal and diabetic placenta. PMID- 11504531 TI - Villous trophoblast: morphometric perspectives on growth, differentiation, turnover and deposition of fibrin-type fibrinoid during gestation. AB - Villous trophoblast growth and deposition of perivillous fibrin-type fibrinoid were examined in human placentas from 10-41 weeks of gestation. The main aims were: (1) to study growth of different trophoblast domains implicated in epithelial turnover (proliferation, differentiation, extrusion, denudation); (2) to test predictions about relationships between fibrinoid deposits and intervillous volume or villous surface area; and (3) to derive baseline data for future studies on complicated pregnancies. Microscopical fields on trichrome stained paraffin sections were selected by systematic random sampling. Volumes were estimated stereologically by point counting and surface areas by intersection counting. Apparent differences were tested by analyses of variance and relationships by regression and contingency table analyses. All compartments increased in absolute volume and/or surface area although not at the same rates. Relative volumes of cytotrophoblast were greater at earlier stages (10-20 weeks) but, due to differential growth, syncytiotrophoblast nuclear aggregation sites (syncytial knots and 'bridges') occupied greater proportions of trophoblast volume and surface near term (37-41 weeks). Fibrinoid volume correlated positively with intervillous volume and villous surface area but, relative to intervillous volume, seemed to increase near term. Findings confirm that the incidence of syncytial knots increases during gestation and contributes to trophoblast thickness variability. Greater relative volumes and surfaces of syncytial 'bridges' are consistent with increased incidences of true intervillous bridges and/or villous branching points. These findings support the notion that fibrinoid deposition during normal gestation is influenced by the quality of vascular perfusion but also emphasize that the villous surface is another important factor. Haemostatic events operate at the maternal surface of trophoblastic epithelium and influence the steady state between coagulation and fibrinolysis. Fibrinoid is deposited at sites of trophoblast de-epithelialization and these arise following trauma (e.g. abruption of intervillous bridges) or during the extrusion phase of normal epithelial turnover. Like knots and bridges, sites of de-epithelialization also expand at a faster rate than overall villous surface area. These and other events in villous development can be interpreted in terms of a coherent concept of epithelial turnover in which proliferation early in gestation is mainly for growth whilst that at later stages is mainly for renewal and repair. PMID- 11504532 TI - Human trophoblast contains an intracellular protein reactive with an antibody against CD133--a novel marker for trophoblast. AB - CD133 is a protein expressed on the cell membrane of a subfraction of haematopoietic stem and progenitor cells, as well as on some epithelial cells. Previously available antibodies against CD133 recognized only the glycosylated protein, localized to membrane protrusions or microvilli. Due to this, immature intracellular stages of the CD133 protein could not be visualized using these antibodies. We describe reactivity of a commercially available antibody against CD133, called AC133-2, with an intracellular protein in trophoblast. Both villous and extravillous cytotrophoblast, as well as syncytiotrophoblast were stained by AC133-2 in cryostat sections of first trimester and term placenta. Villous stroma was not stained. AC133-2 reactivity was seen in methanol-fixed primary trophoblast cells and trophoblast-derived cell lines, and was coexpressed with cytokeratin-7. CD133 messenger RNA was present in trophoblast and trophoblast derived cell lines, but also in cells not displaying any reactivity with CD133 antibodies. AC133-2 recognized a 55-60 kDa protein on Western blots of cell extracts including trophoblast. The exact nature of this protein is not yet understood. However, AC133-2 is applicable as a positive marker for the characterization of all subtypes of trophoblast and for trophoblast cell lines. PMID- 11504533 TI - The spontaneously hypertensive rat fetus, not the mother, is responsible for the reduced amniotic fluid PTHrP concentrations and growth restriction. AB - Intrauterine parathyroid hormone-related protein (PTHrP) concentrations are reduced in association with growth restriction in the spontaneously hypertensive rat (SHR) compared to those of its normotensive control, the Wistar Kyoto (WKY) rat, implicating PTHrP as a pivotal fetal growth factor. The aim of this study was to examine, by embryo cross-transplanation between SHR and WKY, whether the mother, fetus, or both, are responsible for the suppressed SHR amniotic fluid PTHrP. One-day-old SHR embryos were gestated in either an SHR (SHR-in-SHR) or WKY (SHR-in-WKY) surrogate, similarly one-day-old WKY embryos were gestated in either an SHR (WKY-in-SHR) or WKY (WKY-in-WKY) mother. At 20 days gestation, maternal plasma and amniotic fluid samples were collected and assayed for PTHrP concentrations. Data were analysed by two-way ANOVA (mean+/-sem, n=5-9 mothers/group). There were no differences in litter number or maternal plasma PTHrP concentrations. Fetal weight (P< 0.009), fetal/placental weight ratio (P< 0.004) and amniotic fluid PTHrP concentrations (P< 0.001) were lower and amniotic fluid volume (P< 0.0001) was higher with an SHR fetus compared to the WKY fetus irrespective of maternal strain. Thus, the SHR fetus is growth restricted and has suppressed amniotic fluid PTHrP, which are largely determined by the fetus or gestational tissues and are independent of maternal hypertension or maternal PTHrP. We suggest that the low SHR amniotic fluid PTHrP may play a role in the development of SHR growth restriction. PMID- 11504535 TI - The virus-chaperone connection. PMID- 11504534 TI - Isolation and characterization of a bovine blastocyst-derived trophoblastic cell line, BT-1: development of a culture system in the absence of feeder cell. AB - We established a trophoblastic cell line, bovine trophoblast-1 (BT-1), derived from in vitro matured and fertilized blastocyst. While several trophoblastic cell lines have been previously reported using feeder cell, BT-1 could be cultured in the absence of feeder cell. BT-1 was cultured for more than 18 months (over 75 passage) in the absence of feeder cells, using bovine endometrial fibroblast conditioned medium (fibroblast-conditioned medium). We found that the cell growth was accelerated in fibroblast-conditioned medium. In bromodeoxyuridine incorporation analysis, BT-1 cells growth rate in fibroblast-conditioned medium was about two-fold higher than that in conventional medium. Furthermore, fibroblast-conditioned medium accelerated attachment of BT-1 cells to culture dishes following plating. BT-1 showed epithelial morphology and expressed cytokeratin. During continuous culture, cells accumulated fluid under the cell sheet and form dome-like structure that eventually transformed into free floating vesicles. Reverse transcription polymerase chain reaction analysis and immunoblot analysis demonstrated that BT-1 cells expressed interferon-tau as well as placental lactogen (PL). Immunofluorescence analysis demonstrated that a small number of cells were PL-positive, and these cells were binucleate. The BT-1 trophoblastic cell line could serve as a powerful model system for the study of trophoblast cell lineage and proliferation. PMID- 11504536 TI - Full-genome nucleotide sequence of a hepatitis E virus strain that may be indigenous to Japan. AB - We identified hepatitis E virus (HEV) RNA in serum from a Japanese patient with acute hepatitis, who had never been abroad. The full-genome nucleotide sequence of the HEV isolate (JRA1) from this patient was composed of 7227 nucleotides excepting the poly(A) tail and had ORF1 coding for 1703 amino acids (aa), ORF2 coding for 660 aa, and ORF3 coding for 122 aa. This Japanese strain showed approximately 87% nucleotide similarity to human and swine strains reported from the United States, while it had only 73-76% similarity to Asian and Mexican strains. Here we report the characteristics of the HEV-JRA1 isolate, which might be the first example of an indigenous strain(s) of HEV in Japan. PMID- 11504537 TI - No evidence of HIV and SIV sequences in two separate lots of polio vaccines used in the first U.S. polio vaccine campaign. AB - We obtained sealed vials of two different polio vaccine lots, expiration date 1955, which were used in the first U.S. polio vaccine campaign. These early lots were pulled from the market because they contained live infectious poliovirus which caused polio in some of the vaccines. Theoretically, these vaccines could have contained other infectious retroviruses, including HIV. No viral sequences were detected using RT-PCR analyses with primers capable of amplifying chimpanzee SIV and HIV-1-related viruses nor with primers for macaque SIV, sooty mangabey SIV, and HIV-2-related viruses. Poliovirus sequences were readily amplified by RT PCR, suggesting that the technique used would have detected SIV or HIV sequences, if present. PMID- 11504538 TI - An alpha-helical domain within the carboxyl terminus of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) is associated with cell fusion and resistance to heparin inhibition of cell fusion. AB - Previous studies from our laboratory indicated that a 28-amino-acid carboxyl terminal truncation of gB caused extensive virus-induced cell fusion (Baghian et al., 1993, J Virol 67, 2396-2401). We tested the ability of additional truncations and mutations within gB to cause cell fusion in the recently established virus-free cell fusion assay (Turner et al., 1998, J. Virol. 72, 873 875). Deletion of the carboxyl-terminal 28 amino acids of gB (gBDelta28), which removed part of the predicted alpha-helical structure H17b, caused extensive cell fusion. A gB truncation specified by gBDelta36, which removed the entire H17b domain, caused as much cell fusion as the gBDelta28 truncation. Similarly, gB(A874P) containing a substitution of an Ala with Pro within H17b caused cell fusion. Heparin, a gB-specific inhibitor of virus-induced cell fusion, inhibited both wild-type gB and gB(syn3)-mediated cell fusion. In contrast, fusion of cells transfected with gB(Delta28), gB(Delta36), or gB(A874P) was resistant to heparin inhibition of cell fusion. We concluded the following: (1) The predicted alpha helical structure of H17b within the carboxyl terminus of gB is involved in both virus-induced and virus-free cell fusion. (2) Heparin is a specific inhibitor of gB-mediated fusion in both systems. (3) Resistance to heparin inhibition of gB mediated cell fusion is associated with the predicted alpha-helical structure H17b within the carboxyl terminus of gB. PMID- 11504539 TI - Altered biology of adeno-associated virus type 2 and human papillomavirus during dual infection of natural host tissue. AB - Adeno-associated virus (AAV), a common genital virus, may have a "protective" role against human papillomavirus (HPV)-associated cervical cancer. Epidemiological studies indicate a negative correlation between AAV infection and the incidence of cervical cancer. In contrast, HPV is positively associated with cervical cancer. To investigate interactions between these two viruses we used the organotypic "raft" culture system. The raft culture system is capable of supporting the complete HPV life cycle. Raft tissues that were actively replicating HPV were superinfected with AAV type 2 (AAV-2). We observed a multiplicity of infection (m.o.i.)-dependent enhancement and inhibition of HPV DNA replication, concomitant with AAV-2 replication. The data suggest that at low m.o.i. of AAV-2 infection, HPV DNA replication was slightly increased compared to controls and AAV-2 replicated poorly. At high AAV-2 m.o.i., HPV DNA replication was reduced and AAV-2 replicated to high levels. AAV-2 replication was increased in the presence of HPV compared to primary human keratinocyte, squamous cell carcinoma, and HaCat raft cultures infected with AAV-2 alone. These data suggest that HPV may provide types of "enhancer/helper" functions for AAV-2 replication and progeny formation. Infection with AAV-2 had significant effects on epithelial morphology. During infection with low m.o.i. of AAV-2 the epithelium stratified to a greater extent than in controls. With high m.o.i. of AAV-2 infections, tissue cytopathic effects were observed, indicating an additional factor responsible for the effect of AAV-2 on HPV replication and infection. Our results demonstrate a complex interaction between AAV-2, HPV, and skin during dual infection. PMID- 11504540 TI - Effects of alanine cluster mutations in the D12 subunit of vaccinia virus mRNA (guanine-N7) methyltransferase. AB - The (guanine-N7)-methyltransferase domain of the vaccinia virus mRNA capping enzyme is a heterodimer composed of a catalytic subunit D1(498-844) bound to a stimulatory subunit D12. To identify structural elements of the 287-amino-acid D12 subunit that participate in binding and activation of the catalytic subunit, we introduced 12 double-alanine mutations at vicinal residues that are conserved in the D12 homologs of other vertebrate poxviruses. His-tagged D12 mutants were coexpressed in bacteria with the D1(498-544) subunit, and the recombinant D1(498 844)/His-D12 heterodimers were purified. Eight of the mutants (K111A-R112A, N120A N121A, N126A-N127A, F141A-R142A, K223A-D224A, H260A-S261A, E275A-N276A, and R280A R281A) had no significant effect on methyltransferase activity. Three of the mutants (L61A-K62A, F176A-K177A, and F245A-L246A) displayed an intermediate level of cap methylation (35-50% of wild-type activity). Only one mutation, N42A-Y43A, elicited a significant loss of the methyltransferase activation function (<20% of the wild-type activity). Nine of the D12-Ala/Ala proteins were produced individually in bacteria and tested for reconstitution of methyltransferase activity in vitro by mixing with the catalytic subunit. K111A-R112A, N120A-N121A, F176A-K177A, F245A-L246A, and L61A-K62A displayed diminished affinity for the D1 catalytic subunit. N42A-Y43A was uniquely defective in its ability to activate cap methylation by the catalytic subunit. Our results suggest that the methyltransferase activation function of D12, though clearly dependent on the physical interaction with D1, also requires constituents of D12 that are engaged specifically in catalysis. PMID- 11504541 TI - The 3' stem loop of the West Nile virus genomic RNA can suppress translation of chimeric mRNAs. AB - Cis-acting elements that regulate translation have been identified in the 3' noncoding regions (NCRs) of cellular and viral mRNAs. As one means of analyzing the effect on translation of the conserved 3' terminal RNA structure of the West Nile virus (WNV) genome, the translation efficiencies of chimeric mRNAs composed of a CAT reporter gene flanked by viral or nonviral 5' and 3' terminal sequences were compared. In vitro, the WNV 3'(+) stem loop (SL) RNA reduced the translation efficiencies of chimeric mRNAs with either viral or nonviral 5' NCRs, suggesting that a specific 3'-5' RNA-RNA interaction was not involved. In contrast, the 3' terminal sequence of a togavirus, rubella virus, enhanced translation efficiency. The WNV 3'(+)SL reduced translation efficiency both in cis and in trans and of both capped and uncapped chimeric mRNAs. We have previously reported that three cellular proteins bind specifically to the WNV 3'(+)SL RNA. Competition between the WNV 3'(+)SL and the 5' terminus of the chimeric mRNAs for proteins involved in translation initiation could explain the translation inhibition observed. PMID- 11504542 TI - Expression of Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor monocistronic and bicistronic transcripts in primary effusion lymphomas. AB - Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) encodes a G protein-coupled receptor (vGPCR) in open reading frame (ORF) 74, which is homologous to human chemokine receptors. KSHV vGPCR is constitutively active and induces VEGF mediated angiogenesis. Previous studies have shown that ORF 74 is transcribed as part of a bicistronic message containing ORF K14 upstream of ORF 74, with an early lytic pattern of expression. We have now extended these studies by analyzing three different KSHV-positive primary effusion lymphoma (PEL) cell lines and three PEL clinical samples. In addition, we have identified another less abundant monocistronic transcript containing only ORF 74. Both transcripts were identified at low but similar levels in two PEL clinical samples. We evaluated the degree of sequence and functional conservation of ORF74 in three additional PELs and two KS clinical specimens, demonstrating complete identity at the amino acid level among all isolates. While it is expressed as an early lytic transcript in PEL cell lines, in primary clinical PEL samples transcription of KSHV vGPCR can be readily detected. PMID- 11504543 TI - Attenuated and wild-type HIV-1 infections and long terminal repeat-mediated gene expression from plasmids delivered by gene gun to human skin ex vivo and macaques in vivo. AB - Gene expression from HIV-based gene therapy vectors or live-attenuated HIV-1 vaccines requires RNA transcription supported by the HIV-1 promoter, the long terminal repeat (LTR). Delivery of live-attenuated HIV-1 vaccines as plasmid DNA would overcome problems associated with production of attenuated HIV-1 strains. We investigated the expression of reporter plasmids and proviral HIV-1 constructs driven by either the HIV-1 LTR or LTRs with deletions in the U3 enhancer regions. LTR-driven plasmids were inoculated by gene gun into both human epidermis ex vivo and macaques in vivo. The HIV-1 LTR drove reporter gene expression in human and macaque skin, although with 15- to 20-fold less efficiency compared to the immediate-early cytomegalovirus promoter. A deleted LTR derived from a naturally attenuated HIV-1 strain infecting a member of the well-characterized Sydney Blood Bank Cohort of long-term nonprogressors was 5-fold less efficient in expression of the reporter gene compared to wild-type LTR. Delivery of proviral wild-type HIV-1 DNA constructs to human skin resulted in recovery of HIV-1 from cells emigrating from the epidermis, providing an ex vivo model of the infectivity of proviral HIV-1 DNA. However, delivery of proviral HIV-1 DNA containing deletions in either the LTR, Nef, or the secondary viral transcription activator,Vpr, significantly reduced HIV-1 replication in this model. The early coexpression of Tat from a second plasmid did not restore replication. Thus, although attenuated lentiviral vaccines might be deliverable as proviral DNA constructs in primate subjects, significant improvements are needed to enhance the efficiency of this method. PMID- 11504544 TI - Preferential utilization of the perforin/granzyme pathway for lysis of Epstein Barr virus-transformed lymphoblastoid cells by virus-specific CD4+ T cells. AB - In this report, we show that Epstein-Barr virus (EBV)-infected lymphoblastoid cell lines (LCL) express Fas and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor 2 and that LCL are lysed following engagement of these receptors by agonist Fas and TRAIL receptor-specific monoclonal antibodies (MAb). We also show that EBV-specific CD4+ T cells mediate bystander lysis of susceptible targets through both the Fas/Fas ligand (FasL) and the TRAIL pathways, but find that the dominant mechanism of lysis following cognate, HLA class II-restricted recognition of LCL is the perforin/granzyme pathway. Killing of LCL by EBV-specific CD4+ T cells was strongly inhibited by concanamycin A, an agent that elevates granule pH, resulting in accelerated destabilization and degradation of perforin. In contrast, blocking anti-FasL MAb showed only limited inhibition of LCL killing. Blocking anti-TRAIL MAb had no effect on lysis of LCL by EBV-specific CD4+ T cells. We further show that culture of EBV-specific CD4+ T cells in the presence of interleukin 4 markedly abrogates effector cytotoxic function against LCL through direct depletion of intracellular perforin, with no evidence of a Th1 to Th2 shift in patterns of cytokine expression. PMID- 11504545 TI - Nuclear factor 1 family members mediate repression of the BK virus late promoter. AB - BK virus (BKV) is a member of the polyoma virus family that is ubiquitous in humans. Its 5-kb DNA genome consists of a bidirectional promoter region situated between two temporally regulated coding regions. We mapped the transcription initiation site of the major late promoter (MLP) of the archetype strain BKV(WW) to nt 185. We found that it lies within the sequence TGGN6GCCA, a binding site for members of the nuclear factor 1 (NF1) family of transcription factors. Competition electrophoretic mobility shift and immunoshift assays confirmed that NF1 factors present in nuclear extracts of HeLa and CV-1 cells bind to the BKV MLP. Because BKV(WW) grew poorly in tissue culture and failed to express detectable levels of RNA in vitro, SV40-BKV chimeric viruses were constructed to investigate the transcriptional function of this NF-1 binding site. These sequence-specific factors repressed transcription in a cell-free system when template copy number was low. This repression could be relieved by the addition in trans of oligonucleotides containing wild-type, but not mutated, NF1-binding site sequences. SV40-BKV chimeric viruses defective in this NF1-binding site overproduced late RNA at early, but not late, times after transfection of CV-1 cells. Finally, transient expression in 293 cells of cDNAs encoding the family members NF1-A4, NF1-C2, and NF1-X2 specifically repressed transcription from the BKV late promoter approximately 3-, 10-, and 10-fold, respectively, in a DNA binding-dependent manner. We conclude that some members of the NF1 family of transcription factors can act as sequence-specific cellular repressors of the BKV MLP. We propose that titration of these and other cellular repressors by viral genome amplification may be responsible in part for the replication-dependent component of the early-to-late switch in BKV gene expression. PMID- 11504546 TI - Quantitative analysis of herpesvirus sequences from normal tissue and fibropapillomas of marine turtles with real-time PCR. AB - Quantitative real-time PCR has been used to measure fibropapilloma-associated turtle herpesvirus (FPTHV) pol DNA loads in fibropapillomas, fibromas, and uninvolved tissues of green, loggerhead, and olive ridley turtles from Hawaii, Florida, Costa Rica, Australia, Mexico, and the West Indies. The viral DNA loads from tumors obtained from terminal animals were relatively homogeneous (range 2 20 copies/cell), whereas DNA copy numbers from biopsied tumors and skin of otherwise healthy turtles displayed a wide variation (range 0.001-170 copies/cell) and may reflect the stage of tumor development. FPTHV DNA loads in tumors were 2.5-4.5 logs higher than in uninvolved skin from the same animal regardless of geographic location, further implying a role for FPTHV in the etiology of fibropapillomatosis. Although FPTHV pol sequences amplified from tumors are highly related to each other, single signature amino acid substitutions distinguish the Australia/Hawaii, Mexico/Costa Rica, and Florida/Caribbean groups. PMID- 11504547 TI - The complete DNA sequence of the Ectocarpus siliculosus Virus EsV-1 genome. AB - The Ectocarpus siliculosus Virus-1, EsV-1, is the type-species of a genus of Phycodnaviridae, the phaeoviruses, infecting marine filamentous brown algae. The EsV-1 genome of 335,593 bp contains tandem and dispersed repetitive elements in addition to a large number of open reading frames of which 231 are currently counted as genes. Many genes can be assigned to functional groups involved in DNA synthesis, DNA integration, transposition, and polysaccharide metabolism. Furthermore, EsV-1 contains components of a surprisingly complex signal transduction system with six different hybrid histidine protein kinases and four putative serine/threonine protein kinases. Several other genes encode polypeptides with protein-protein interaction domains. However, 50% of the predicted genes have no counterparts in data banks. Only 28 of the 231 identified genes have significant sequence similarities to genes of the Chlorella virus PBCV 1, another phycodnavirus. To our knowledge, the EsV-1 genome is the largest viral DNA sequenced to date. PMID- 11504548 TI - A novel inducible expression system to study transdominant mutants of HIV-1 Vpr. AB - In this study, an episomal system for ecdysone-inducible gene expression was developed. Human embryonic kidney 293 cells (293VE) expressing a heterodimer of modified ecdysone and retinoid X receptors and the Epstein-Barr nuclear antigen-1 were screened. Plasmids containing the EBV replication origin, oriP, and the ecdysone-response element could replicate and persist in 293VE cells to inducibly express luciferase or Vpr. The induction level, tested with luciferase reporter plasmid, varied among cell lines from 254- to 2056-fold. In one highly inducible cell line, HIV-1 Vpr was expressed well and caused G2 cell cycle arrest in the presence of the inducer, while in the absence of the inducer, no Vpr protein or cell cycle arrest could be detected. Using different selection markers, HIV-1 Vpr was coexpressed with Vpr mutants defective in phosphorylation at Ser79 and G2 cell cycle arrest activity. These Vpr mutants were transdominant to wild-type Vpr for G2 cell cycle arrest activity, but did not alter wild-type Vpr phosphorylation. It is likely that the transdominant mutants and wild-type Vpr compete for a downstream target(s) of G2 cell cycle arrest. PMID- 11504549 TI - Mutations at amino acid positions 63, 189, and 396 of human immunodeficiency virus type 1 reverse transcriptase (RT) partially restore the DNA polymerase activity of a Trp229Tyr mutant RT. AB - Trp229 is part of the nonnucleoside reverse transcriptase inhibitor (NNRTI) binding pocket of HIV-1 reverse transcriptase (RT). It is also an important constituent of the so-called "primer grip." Using a recombinant virus assay, we tried to obtain recombinant virus containing a Trp229Phe or a Trp229Tyr mutation in its RT. Previous studies already established the very low DNA polymerase activities of both the Trp229Phe and the Trp229Tyr mutant RT enzymes. We were able to obtain a Trp229Tyr but not a Trp229Phe mutant virus. However, in addition to the Trp229Tyr mutation this mutant virus also contained an Ile63Met, a Val189Ile, and a Glu396Gly mutation in its RT. When we evaluated the quadruple mutant virus for sensitivity/resistance against a variety of NNRTIs, no significant difference with the sensitivity/resistance profile of the single Trp229Tyr mutant RT enzyme could be observed. We found that the three additional mutations partly restored the low RNA- and DNA-dependent DNA polymerase activities of the Trp229Tyr mutant enzyme. Kinetic analysis revealed that both template/primer binding and dNTP incorporation are affected by the Trp229Tyr mutation. Our findings demonstrate that a mutation at position 229 is unlikely to occur under NNRTI drug pressure due to the poor catalytic activity of the singly mutated RT and the favorable drug sensitivity profile of the mutated enzyme/viruses in both the absence and the presence of the compensatory mutations. Therefore, amino acid position 229 may be regarded as an excellent amino acid target within the NNRTI pocket for rational drug design. PMID- 11504550 TI - Subcellular distribution of the foot-and-mouth disease virus 3A protein in cells infected with viruses encoding wild-type and bovine-attenuated forms of 3A. AB - Picornavirus infection induces the proliferation and rearrangement of intracellular membranes in response to the synthesis of nonstructural proteins, including 3A. We have previously shown that changes in 3A are associated with the inability of a Taiwanese strain of foot-and-mouth disease virus (FMDV) (OTai) to grow in bovine cells and cause disease in cattle, although the virus grows to high titers in porcine cells and is highly virulent in pigs (C. W. Beard and P. W. Mason, 2000, J. Virol. 74, 987-991). To study if differences in the distribution of 3A could account for the species specificity of OTai, we compared the localization of the OTai 3A with a bovine-virulent 3A (serotype A12) in keratinocytes prepared from the tongues of cattle and pigs. Following either infection of keratinocytes or transfection with 3A we were unable to discern differences in 3A distribution in either species of keratinocyte, independent of the strain of virus (or 3A) utilized. In both cell types, 3A distributed in a pattern that overlapped with an endoplasmic reticulum (ER) marker protein, calreticulin (CRT). Furthermore, although FMDV infection or transfection with 3A did not result in a gross redistribution of CRT, both virus infection and 3A transfection disrupted the Golgi. Other picornaviruses that disrupt Golgi function are sensitive to brefeldin A (BFA), a fungal metabolite that interferes with retrograde transport between the Golgi and the ER. Interestingly, BFA has little effect on FMDV replication, suggesting that FMDV may acquire cellular membranes into its replication complexes in a manner different from that of other picornaviruses. PMID- 11504551 TI - Impaired microglial activation in the brain of IL-18-gene-disrupted mice after neurovirulent influenza A virus infection. AB - Knockout of the interleukin-18 (IL-18) gene predisposed mice to impaired clearance of neurovirulent influenza A virus-infected neurons from the brain. In wild-type mice, IL-18 molecule-producing microglia/macrophages emerged in virally attacked regions as early as day 3 after infection. Microglial transformation into macrophages culminated at day 7 to 9, with upregulated expression of Iba1, a novel calcium-binding protein that controls phagocytic functions of microglia/macrophages. In IL-18-/- mice, microglial transformation was interrupted with reduced Iba1 expression. Interferon-gamma (IFN-gamma) immunopositive neurons appeared in and around virally invaded regions in wild type mice, peaking in number at day 7, whereas such cells were barely detected in IL-18-/- mice. Stereotaxic microinjection of recombinant IFN-gamma triggered microglial transformation in IL-18-/- mice and upregulated Iba1 expression, leading to effective eradication of virally infected neurons. Collectively, these results suggest that IL-18 plays a key role in activating microglial functions directed against the influenza virus infection by inducing neuronal IFN-gamma in the brain parenchyma. PMID- 11504552 TI - Phosphorylation of Marburg virus VP30 at serines 40 and 42 is critical for its interaction with NP inclusions. AB - The Marburg virus (MBGV) nucleocapsid complex is composed of four viral proteins (NP, L, VP35, and VP30) and the negative-strand nonsegmented genomic RNA. NP, L, and VP35 are functionally conserved among the order Mononegavirales, whereas VP30, a phosphoprotein, represents a filovirus-specific nucleocapsid protein. In the present paper, we have characterized the localization and function of VP30 phosphorylation. The main phosphorylation sites are represented by seven serine residues in the region of amino acid 40 to 51 of VP30. Additionally, trace amounts of phosphothreonine were detected. Substitution of serine residues 40 and 42 by alanine abolished the interaction of VP30 with NP-induced inclusion bodies, which contain nucleocapsid-like structures formed by NP. Substitution of the other phosphoserine residues had little effect on this interaction. Replacement of the introduced alanine residues 40 and 42 by aspartate restored the interaction between VP30 and the NP inclusions pointing to the importance of negative charges at these particular positions. PMID- 11504553 TI - A 10-kDa structural protein of porcine reproductive and respiratory syndrome virus encoded by ORF2b. AB - The major structural proteins of porcine reproductive and respiratory syndrome virus (PRRSV) are derived from ORFs 5, 6, and 7. Western blots of sucrose gradient-purified virions and PRRSV-infected MARC-145 cells, probed with immune pig serum, showed the presence of an additional 10-kDa protein. Nucleotide sequence analysis of North American PRRSV isolate SDSU-23983 revealed a small ORF within ORF2, named ORF2b, which, when translated, produced a 73-amino-acid nonglycosylated protein. Recombinant 2b protein expressed by a baculovirus clone, AcVR2, comigrated with the 10-kDa virus-associated protein. The loss of 10-kDa protein immunoreactivity after absorption of immune sera with lysates from AcVR2 infected insect cells demonstrated that the 2b and 10-kDa proteins are immunologically similar. Immunoblots were also used for the detection of anti-2b activity in serum samples from experimentally infected adult pigs. Antibodies against PRRSV were apparent by 14 days postinfection, followed by anti-2b activity and serum neutralizing activity. The putative ORF2b start codon is only 6 nucleotides downstream of the adenine of the ORF2a start codon. The expression of ORF2a and 2b as enhanced green fluorescent fusion proteins showed that both proteins were translated; however, the ORF2b was preferentially expressed. These results suggest that the 2b protein is virion associated and the principal product of ORF2. PMID- 11504554 TI - Molecular characterization of the polymerase gene and genomic termini of Nipah virus. AB - In 1998, Nipah virus (NV) emerged in peninsular Malaysia, causing fatal encephalitis in humans and a respiratory disease in swine. NV is most closely related to Hendra virus (HV), a paramyxovirus that was identified in Australia in 1994, and it has been proposed that HV and NV represent a new genus within the family Paramyxoviridae. This report describes the analysis of the sequences of the polymerase gene (L) and genomic termini of NV as well as a comparison of the full-length, genomic sequences of HV and NV. The L gene of NV is predicted to be 2244 amino acids in size and contains the six domains found within the L proteins of all nonsegmented, negative-stranded (NNS) RNA viruses. However, the GDNQ motif found in most NNS RNA viruses was replaced by GDNE in both NV and HV. The 3' and 5' termini of the NV genome are nearly identical to the genomic termini of HV and share sequence homology with the genomic termini of other members of the subfamily Paramyxovirinae. At 18,246 nucleotides, the genome of NV is 12 nucleotides longer than the genome of HV and they have the largest genomes within the family Paramyxoviridae. The comparison of the structures of the genomes of HV and NV is now complete and this information will help to establish the taxonomic position of these novel viruses within the family Paramyxoviridae. PMID- 11504555 TI - The involvement of a stress-activated pathway in equine influenza virus-mediated apoptosis. AB - We have shown elsewhere that equine-2 influenza virus (EIV; subtype H3N8) induced pronounced cell death in infected cells through apoptosis as demonstrated by DNA fragmentation assay and a combined TUNEL and immunostaining scheme. In this study, we investigated the mechanism of EIV-mediated cytotoxicity on a permissive mammalian epithelial cell line, Madin-Darby canine kidney (MDCK) cells. EIV infection increased the cellular levels of oxidative stress and c-Jun/AP-1 protein (which is known to be affected by oxidative stress), as well as its DNA binding activity. Increased production of TGF-beta1, an inducer of c-Jun N terminal kinase or stress-activated protein kinase (JNK/SAPK) activation, was also detected in EIV-infected MDCK cells. It has been reported that TGF-beta may initiate a signaling cascade leading to JNK/SAPK activation. Addition of c-Jun antisense oligodeoxynucleotide, antioxidant N-acetyl-cysteine (NAC), JNK/SAPK inhibitor carvedilol, or TGF-beta-neutralizing antibody effectively blocked c Jun/AP-1 upregulation and TGF-beta1 production mediated by EIV infection. These treatments also attenuated EIV-induced cytopathogenic effects (CPE) and apoptosis. Our results suggest that a stress-activated pathway is involved in apoptosis mediated by EIV infection. It is likely that EIV infection turns on the JNK/SAPK cascade, which modulates the activity of apoptosis-promoting regulatory factor c-Jun/AP-1 and epithelial growth inhibitory cytokine TGF-beta. PMID- 11504556 TI - Regulation of human papillomavirus type 1 replication by the viral E2 protein. AB - The E1 and E2 proteins encoded by papillomaviruses are required for viral replication. Earlier studies have shown that the viral E2 protein plays an important role in replication by targeting the E1 helicase to the origin of replication (ori). We have previously shown that the E1 protein of human papillomavirus (HPV) type 1 is sufficient for the in vivo replication of ori plasmids, although the E2 protein stimulates replication. In this study, we have further analyzed the role of the E2 protein in HPV-1 replication. The optimal ori of HPV-1 contains one putative E1 binding site (E1BS) and two putative E2 binding sites, E2BS-3 and E2BS-4. Plasmid pori171, containing the optimal ori, replicates to much higher levels than plasmid pori312, which includes an additional upstream E2 binding site, E2BS-2, located 75 nucleotides upstream of E2BS-3. To study the possible role of E2BS-2 and other upstream sequences in E2-dependent downregulation of replication, transient replication analysis was done in the presence of increasing levels of the E2 protein. Interestingly, inhibition of pori312 replication was more severe at higher levels of E2, suggesting that this protein may also negatively regulate HPV-1 replication. Deletion of sequences from pori312 containing an additional putative E2BS, E2BS-2A, relieved the repression of replication to a significant extent, while replacement of E2BS-2 with a different sequence of the same length had a modest effect. These results suggest that E2BS-2A plays a major and E2BS-2 a minor role in the negative regulation of HPV-1 replication at high E2 levels. Electrophoretic mobility-shift assays showed that the purified E2 protein bound with high affinity to E2BS-3 and weakly to the other putative E2BSs located within the viral long control region. EMSA using various ori fragments showed the formation of multiple E2-DNA complexes which likely represent binding of E2 to multiple E2BSs present within the HPV-1 ori. Our data are consistent with the assembly of ori-protein complexes at high E2 levels that are impaired for replication and further suggest that E2 may regulate HPV-1 replication by a mechanism involving interaction between the E2 protein bound to E2BSs at a distance. PMID- 11504557 TI - The nucleotide sequence and genomic organization of Citrus leaf blotch virus: candidate type species for a new virus genus. AB - The complete nucleotide sequence of Citrus leaf blotch virus (CLBV) was determined. CLBV genomic RNA (gRNA) has 8747 nt, excluding the 3'-terminal poly(A) tail, and contains three open reading frames (ORFs) and untranslated regions (UTR) of 73 and 541 nucleotides at the 5' and 3' termini, respectively. ORF1 potentially encodes a 227.4-kDa polypeptide, which has methyltransferase, papain-like protease, helicase, and RNA-dependent RNA polymerase motifs. ORF2 encodes a 40.2-kDa polypeptide containing a motif characteristic of cell-to-cell movement proteins. The 40.7-kDa polypeptide encoded by ORF3 was identified as the coat protein. The genome organization of CLBV resembles that of viruses in the genus Trichovirus, but they differ in various aspects: (i) in trichoviruses ORF2 overlaps ORFs 1 and 3, whereas in CLBV, ORFs 2 and 3 are separated and ORFs 1 and 2 overlap in one nucleotide; (ii) CLBV gRNA and CP are larger than those of trichoviruses; and (iii) the CLBV 3' UTR is larger than that of trichoviruses. Phylogenetic comparisons based on CP amino acid signatures clearly separates CLBV from trichoviruses. Also contrasting with trichoviruses, CLBV could not be transmitted to Chenopodium quinoa Willd. Considering these singularities, we propose that CLBV should be included in a new virus genus. PMID- 11504558 TI - Correlation of genetic variability with safety of mumps vaccine Urabe AM9 strain. AB - The Urabe AM9 strain of mumps vaccine live is known for its genetic instability and some vaccines derived from this strain were withdrawn from the market due to an excessive number of vaccine-associated parotitis and meningitis cases. To identify the molecular basis of this instability, we determined complete nucleotide sequences of several stocks of the Urabe strain used for vaccine production by different manufacturers and of two clinical isolates from cases of vaccine-associated meningitis. In contrast to previously published studies relating the Lys335 --> Glu mutation in the viral HN gene with neurovirulence of mumps virus, we could not confirm any association of this mutation with the safety of mumps vaccine. Each of the three vaccine stocks studied had its own characteristic profile of mutations that was identified by cDNA sequencing and quantitated by mutant analysis by PCR and restriction enzyme cleavage. Determination of the mutational profile of mumps vaccine lots could allow vaccine manufacturers to characterize seed viruses and monitor the consistency of vaccine production to prevent emergence of virulent revertants. PMID- 11504559 TI - The relationship between the L1 and L2 domains of the insulin and epidermal growth factor receptors and leucine-rich repeat modules. AB - BACKGROUND: Leucine-rich repeats are one of the more common modules found in proteins. The leucine-rich repeat consensus motif is LxxLxLxxNxLxxLxxLxxLxx- where the first 11-12 residues are highly conserved and the remainder of the repeat can vary in size Leucine-rich repeat proteins have been subdivided into seven subfamilies, none of which include members of the epidermal growth factor receptor or insulin receptor families despite the similarity between the 3D structure of the L domains of the type I insulin-like growth factor receptor and some leucine-rich repeat proteins. RESULTS: Here we have used profile searches and multiple sequence alignments to identify the repeat motif Ixx-LxIxx-Nx-Lxx Lxx-Lxx-Lxx- in the L1 and L2 domains of the insulin receptor and epidermal growth factor receptors. These analyses were aided by reference to the known three dimensional structures of the insulin-like growth factor type I receptor L domains and two members of the leucine rich repeat family, porcine ribonuclease inhibitor and internalin 1B. Pectate lyase, another beta helix protein, can also be seen to contain the sequence motif and much of the structural features characteristic of leucine-rich repeat proteins, despite the existence of major insertions in some of its repeats. CONCLUSION: Multiple sequence alignments and comparisons of the 3D structures has shown that right-handed beta helix proteins such as pectate lyase and the L domains of members of the insulin receptor and epidermal growth factor receptor families, are members of the leucine-rich repeat superfamily.